PMID- 18380129 TI - Age-related changes in top-ten men's U.S. Masters 50-m freestyle swim times as a function of finishing place. AB - This study described age-related changes in swim time as a function of top-ten finishing place for Masters swimmers. Previous research has documented increases in 50-m freestyle swim times for men beyond age 35 using aggregated top-ten times. The present study examined the relationship between age and swim time, taking into account the potential influence of finishing place in the top-ten. Data were retrieved from the United States Masters Swimming database for male top ten finishers in the 50-m short course freestyle for the years from 1993 to 2002. Data from each year were analyzed using linear regression procedures with Age, Square of age, and Finishing place as predictor variables and Swim time as the dependent variable. Analysis indicated that, when Finishing place was considered, a more gradual increase in Swim time as a function of age was seen for finishing places closer to first place than for those closer to tenth place. PMID- 18380130 TI - Bust size and hitchhiking: a field study. AB - To test the effect of a woman's bust size on the rate of help offered, 1200 male and female French motorists were tested in a hitchhiking situation. A 20-yr.-old female confederate wore a bra which permitted variation in the size of cup to vary her breast size. She stood by the side of a road frequented by hitchhikers and held out her thumb to catch a ride. Increasing the bra-size of the female hitchhiker was significantly associated with an increase in number of male drivers, but not female drivers, who stopped to offer a ride. PMID- 18380131 TI - Volatility kills. PMID- 18380132 TI - Digital diet. PMID- 18380133 TI - Dropping acid. PMID- 18380134 TI - Catching no rays. PMID- 18380135 TI - Wipeout? PMID- 18380137 TI - Let the games begin! PMID- 18380136 TI - Cellulose success. PMID- 18380138 TI - Keys to climate protection. PMID- 18380139 TI - The economist has no clothes. PMID- 18380140 TI - Wag the dog. PMID- 18380141 TI - The color of plants on other worlds. PMID- 18380142 TI - Regrowing human limbs. PMID- 18380143 TI - Reclaiming the Aral Sea. PMID- 18380144 TI - Rulers of light. PMID- 18380145 TI - The doping dilemma. PMID- 18380146 TI - Carbon wonderland. PMID- 18380147 TI - Detecting nuclear smuggling. PMID- 18380148 TI - At the edge of life's code. PMID- 18380149 TI - Weapons revealed. PMID- 18380150 TI - Do cosmic rays cause lightning? PMID- 18380151 TI - Does "spring fever" exist? PMID- 18380152 TI - Identification of genes for two major sialoglycoproteins, glycophorin A and glycophorin C in canine red cell membranes. AB - Glycophorins are the major sialoglycoproteins in red blood cell membranes, possessing various physiological and pathological roles. We examined membrane glycoproteins in canine red cells and cloned cDNAs for two major glycophorins, glycophorins A (GPA) and C (GPC) from bone marrow cells. Periodic acid-Schiff staining and immunoblotting analyses showed that canine red cell membranes contained several glycoproteins immunoreactive to an anti-bovine GPC antibody, whereas the most abundant sialoglycoproteins, the candidates for GPA, did not react with an anti-human GPA antibody. The amino acid sequences of the extracellular domains of GPA and GPC had no significant homology to those from other mammalian species, including humans, and had O-linked and/or N-linked glycosylation sites. On the other hand, the C-terminal cytoplasmic domain and/or the transmembrane helices of GPA and GPC were conserved among species, indicating some functional significance of those regions in red cell membranes that include dimerization of GPA in the membrane-spanning region, and association of GPC with membrane skeletal proteins through binding with protein 4.1 and p55 in the cytoplasmic domain. These findings provide insights for clinical studies to evaluate the involvement of GPA and GPC in the pathogenesis of red cell diseases. PMID- 18380153 TI - Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus. AB - The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKK(250) On SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV. PMID- 18380154 TI - Flow cytometry to evaluate the level of Babesia gibsoni parasitemia in vivo and in vitro by using the fluorescent nucleic acid stain SYTO16. AB - In the present study, we employed flow cytometry to evaluate the level of parasitemia of Babesia gibsoni infecting canine erythrocytes in vivo and in vitro by using fluorescent nucleic acid staining. Peripheral blood samples from a B. gibsoni-infected dog and cultured B. gibsoni parasitizing in canine erythrocytes were stained with a membrane-permeable fluorescent nucleic acid stain, SYTO16. In this study, we utilized normal canine erythrocytes (LK erythrocytes) and canine erythrocytes containing high concentrations of potassium, reduced glutathione, and some free amino acids (HK erythrocytes) as host cells for culture. Parasitized cells in vive were discriminated completely from unparasitized cells and a correlation (r = 0.998) between the percentage of SYTO16-positive cells and parasitemia in vivo was observed. On the other hand, erythrocytes in vitro could not be divided clearly into parasitized and unparasitized cells. However, when LK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was almost the same as, and was well correlated (r = 0.932) with, the level of parasitemia. When HK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was almost half of, but was correlated (r = 0.982) with, the level of parasitemia. Therefore, we attempted to observe the changes in the percentage of parasitized cells after treatment with antiprotozoal drug or mitochondria inhibitors by using flow cytometry. The changes in the percentage of SYTO16-positive cells corresponded well with the changes of the level of parasitemia when the parasites in HK erythrocytes were cultured with each compound. The present results suggest that flow cytometric detection using SYTO16 is a rapid and reliable method for monitoring parasitemia both in vive and in vitro. PMID- 18380155 TI - [Trends in surgery of lung cancer]. PMID- 18380156 TI - [Bilateral pleural effusion--a complication of central venous catheterization--a case review]. AB - The authors present a case review of a 30-year old patient, who developed central pleural effusion, a complication related to central venous catheterization and the consequent use of the central venous line. The authors aim to highlight potential complications of the method. The interesting feature of this case is the fact, that no apparent mistakes in the venous line introduction or its later use have been recorded. PMID- 18380157 TI - [Intra-aortic balloon counterpulsation in cardiac surgery patients--experiences of Department of Cardiac Surgery, Charles University, Faculty of Medicine in Hradec Kralove, University Hospital Hradec Kralove, Czech Republic]. AB - AIM: To evaluate the use of intra-aortic balloon pump (IABP) at the Department of Cardiac Surgery, Charles University, Faculty of Medicine in Hradec Kralove, University Hospital Hradec Kralove and identifying the complications of this method and their frequency in long-term follow-up. METHOD: Retrospective analysis of the outcome of IABP use. From September 1994 to September 2007, 10,024 cardiosurgery operations were performed at the Department of Cardiac Surgery, University Hospital Hradec Kralove, Czech Republic. IABP was applied in 363 cases (3.6%). Therapeutic indication for initiation of IABP at our department was the syndrome of low cardiac output in the postoperative period and during operation (difficulty in weaning from cardio-pulmonary bypass) and severe acute ischemic changes of myocardium refractory to pharmacotherapy. Prophylactic IABP was elective introduction of support before cardiac surgery in hemodynamically stable high risk patients. OUTCOME: From the group of 363 counterpulsated patients 192 were succesfully treated (52.9%), 171 patients (47.1%) died in consequence of cardiogenic shock and multiorgan failure. In 20 cases (5.5%) the IABP was introduced before the operation. Complications were observed in 61 patients (16.8%). Vascular complications were found in 27 cases (7.4%) including ischemic changes of the limb (14 cases 3.9%), significant bleeding occurred at the site of puncture (7 cases 1.9%), dissection of the femoral and iliac arteries (2 cases- 0.55%), perforation of the iliac artery (1 case--0.3%). In 1 case (0.3%) the balloon was removed for intramural hematoma of the descending aorta without dissection. In 2 cases (0.55%) the balloon was led into the venous system. Thrombocytopenia occurred in our group in 23 patients (6.3%). Technical complications were observed in 7 cases (1.9%). None from our group of couterpulsated patinets had infectious complication. CONCLUSION: IABP is an effective and clinically verified mechanical cardiac support. Our results are similar to other studies. PMID- 18380158 TI - [A post-myocardial infarction interventricular septal defect. A case review]. AB - In this case review, the authors describe a case of a mechanical complication of acute myocardial infarction--an interventricular septal defect. The surgery could be postponed, to allow for a safer and technically simpler procedure a month after the onset of the heart attack and of the ventricular septal defect. PMID- 18380159 TI - [Rupture of the interventricular septum after the blunt trauma of thorax]. AB - Authors present the case of little patient with the dissection, pseudoaneurysm and finally a rupture of the interventricular septum after the blunt thracic injury. The patient was smitten as a pedestrian by a car and during the whole period of her stay in the hospital she was showing signs of circulatory instability. Due to the current intraabdominal injuries this circulatory decompensation was first assigned to hemoperitoneum, for which the girl was operated on about 3 hours after admission. Nevertheless, even after the abdminal cavity check, after the treatment of supreficial liver lacerations and intensive volume resuscitation the patient showed signs of insufficiency. Diagnosis was finally determined on the base of the transthoracic echocardiography (TTE), which proved the traumatic rupture of interventricular septum. The operation followed correcting the defect, which was performed with a good result according to the TTE postoperatively. Nevertheless, 27 hours after the admission the patient died due to the electromechanical dissociation. In the discussion the authors then evoke a number of papers concerning the same topic. PMID- 18380160 TI - [Benefits of intraoperative n. laryngeus recurrens neuromonitoring in thyroid surgery]. AB - INTRODUCTION: N. laryngeus reccurens injury is a serious complication in thyroid surgery. Nerve visualization is a procedure preventing its traumatization. The visualization with additional intraopertive neuromonitoring results in further reduction of the n. laryngeus reccurens injury rates. AIM: Verification of the intraoperative neuromonitoring effects on reduction of the n. laryngeus reccurens injury rates. MATERIAL, METHODOLOGY: 309 operated patients were included in the intraoperative neuromonitoring prospective study. A total of 295 patients were included in the study and intraoperative monitoring was performed per protocol. RESULTS: Out of the total of 295 subjects, 1 patient sufferred from permanent paresis, ie. 0.33%, nerves at risk 0.2%. 6 subjects sufferred from transient pareses, i.e. 2.03%, nerves at risk 1.2%, with a prevailing 8-week recovery period of the ORL findings. CONCLUSION: Intraoperative neuromonitoring in thyroid surgery is yet another option for reduction of the n. laryngeus reccurens intraoperative injury rates. PMID- 18380161 TI - ["Tension Free" method in inguinal hernioplasty using CHS 100 mesh--our experience]. AB - The authors present their experience with "Tension Free" method for hernia surgery, using a CGS 100 mesh. They highlight pros and cons of the described method. PMID- 18380162 TI - [Haemobilia, the rare case of bleeding into GIT]. AB - AIM OF THE WORK: Bleeding into GIT is still serious problem with not low lethality. Especially, acute bleeding needs fast diagnostics and treatment. The aim of our work is to present the rare case of bleeding into GIT--haemobilia. GROUP OF THE PATIENTS: There is presented casuistics of the patient who was successfully treated on our department for bleeding from erosion of cystic artery as a consequence of acute cholecystitis. CONCLUSION: Nowadays there is dominantly preferred endoscopic approach in diagnostics and treatment of the bleeding into GIT. But surgical approach is in some cases the only one method in treatment of these complications. PMID- 18380163 TI - [Spontaneous rupture of ductus choledochus in acute pancreatitis--a case report]. AB - Spontaneous perforation of biliary tree is a rare disease. There are only a few case- reports published in the literature. Due to little success in the preoperative diagnosis, surgical treatment is delayed, which in a high percentage results into fatal complications. Surgeons' opinions of operative treatment in such cases are controversial, but the primary choice is decompressing drainage of biliary tree. Authors of this report present a case of 71 years old woman admitted to the second surgical clinic with the diagnosis of acute pancreatitis. Because of developing signs of peritoneal irritation, the patient was referred for a surgical procedure with the finding of biliary peritonitis and the rupture of ductus choledochus. In the above case, the diagnosis was made peroperatively, where the surgical treatment consisted of drainage of ductus choledochus by Kehr T drain and cholecystestomy by inserting Malecoat catheter into gallbladder. Postoperative period was without complications, the T drain was removed on the 28th day after operation and Malecoat catheter was also removed two days later. Three years after the operation, the patient is in good physical condition with free bile passage according to ERCP. The aim of this report is to bring the attention to this rare, but more importantly serious cause of origin of sudden abdominal event, in order to increase the interception rate concerning preoperative periods and consequent satisfactory treatment. PMID- 18380164 TI - [Our experience with radiofrequency--assisted liver resections for hepatic tumors]. AB - INTRODUCTION: A technique for liver resection, utilising radiofrequency-induced coagulation necrosis is considered the latest trend in liver surgery. The authors present their clinical experience with this promising liver resection technique. MATERIAL AND METHODOLOGY: From January 2006 to September 2007, a total of 24 patients underwent radiofrequency-assisted liver resections in the Ostrava FN (Faculty Hospital) Surgical clinic. Liver malignancies (primary or secondary) were resected using open surgery or laparoscopy. The data were analyzed to assess the new technique's outcomes, including intraoperative blood loss, transfusion requirement, morbidity and mortality rates. All the patients were followed up prospectively in regular intervals to assess treatment effectivity, complication rate and the disease relaps rate. RESULTS: Radiofrequency-assisted liver resection was utilised to resect a total of 38 tumors in 24 patients. The mean tumor size was 38 cm. The authors performed 5 major liver resections (> 2 segments) and 19 minor liver resections (nonanatomical resections, segmentectomies). The mean total operation time was 190 minutes, the mean liver resection time was 65 minutes. The mean intraoperative blood loss was 384 ml, however, in the last 10 subjects it was only 70 ml. Four subjects (16.7%) received blood transfusions during the perioperative period, in particlar those, who underwent major resections. In four patients (16.7%), postoperative complications were recorded, however, without lethal outcomes. The mean duration of hospitalization was 11.7 days. Based on the histological examination of the resecates, R0 resections were performed in 91.7% of patients. CONCLUSION: Radiofrequency-assisted liver resection is a safe, well- tolerated and effective liver resection technique. It is associated with minor blood loss, it results in reduced morbidity and mortality rates and may be used in both minor or major liver resections. PMID- 18380165 TI - [Surgical management of proximal humeral fractures using the Hackethal (Zifko) method--a case review]. AB - BACKGROUND: Fractures of the proximal humerus actually represent 5% of all fractures and 45% of all humeral fractures with the highest incidence in women over 60 years of age. The outcomes of the greatest concern at these patients (pain, function of the shoulder and activities of daily living) get worse with age, osteoporosis, grading of fracture type and of initial fracture displacement. According to the literature (Evidence Based Medicine--EBM level II-III) operative treatment of displaced fractures reduces pain and need for assistance in activities of daily living, but open reduction with internal fixation by conventional screws and plates was connected with implant loosening, infection and avascular necrosis, whereas closed reduction with different methods of miniinvasive percutaneous stabilisation is threatened by primary malreduction. AIM: Evaluation of clinical and functional outcomes and analysis of results of proximal humerus fractures treated by closed or percutaneous reduction and intramedullary fixation by means of bundle of Kirschner wires after Zifko. DESIGN: Retrospective descriptive study--case serie. MATERIAL AND METHODS: Within January 1, 2005 - December 31, 2005 there were 87 patients with 87 two- and three fragments fractures of proximal humerus (according to Neers classification) operated at our institution, from which 76 by the method after ZiFko. 36 patients from these 76 came to final evaluation (follow-up rate 47%). AO fracture types were: A2-36%, A3-33%, B1-19%, C1-12%. RESULTS: The resulting Constant-Murley (CM) score reached in mean 89 points with 89% of excellent--good functional results. Complications were recorded in 30% of cases and were represented mainly by proximal migration of K-wires. Avascular necrosis of humeral head occured in two cases (5.5%). CONCLUSION: In spite of low follow-up rate we conclude, that the method of closed reduction and intramedullary fixation of two- and three-part fractures of proximal humerus after Zifko offers above-average final results with acceptable rate of complications, mostly not severe ones. The essential pre condition of good result is proper reduction--closed of percutaneous one. On the contrary, suboptimal results correlate with primary and secondary malreductions. The incidence of latter increases in intraarticular fracture types with small fragment of head and thus with insuficient implant retention. To evaluate the merit of the method in relation to angle-stable extra-/or intramedullary implants, the prospective trials are needed. PMID- 18380166 TI - Effects of aluminum on insulin-like growth factor I levels and antioxidant status. AB - BACKGROUND: The aim of this research was to investigate the toxic effects of aluminum (Al) on plasma insulin-like growth factor I (IGF-I) levels and on the liver, the main production site of IGF-I. In addition, we analyzed the influence of Al on liver malondialdehyde (MDA) and glutathione (GSH) levels, and how the antioxidant vitamin E (vit E) affects the altered levels of these parameters. METHODS: Adult male rats (n = 28) were randomly divided into the following four groups: Al alone, Al + vit E, vit E alone, and untreated control group. The Al group received 1 mg/200 g body weight of aluminum sulfate (AlSO4) thrice weekly for two weeks. The Al + vit E group received the same dose of AlSO4 plus 100 mg/kg of vit E once daily. The Vit E group received a daily dose of vit E alone. Control animals received physiologic saline daily. RESULTS: Liver GSH levels were decreased in the Al group but recovered with vit E administration. Liver IGF-I levels significantly decreased in the Al group compared with the control. With the use of vit E, the liver IGF-I levels increased, but this increase was not statistically significant. CONCLUSIONS: The results of this study show that plasma and liver IGF-I levels decrease with Al use. Also liver GSH levels decreased with Al while this recovered with vit E use together with Al. PMID- 18380167 TI - Effect of l-ascorbic acid on antioxidant defense system in testes of albino rats exposed to nickel sulfate. AB - We studied the effect of oral supplementation with L-ascorbic acid (50 mg/100 g body weight) on nickel sulfate (2.0 mg/100 g body weight, i.p.) induced lipid peroxidation in the testes of Wister strain male albino rats. Testicular lipid peroxide and glutathione (GSH) levels and the activities of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were estimated. Nickel sulfate treatment significantly increased the level of testicular lipid peroxide and decreased all antioxidant enzymes activities and GSH concentration. Simultaneously treatment of L-ascorbic acid exhibited a possible protective role on the toxic effect of nickel sulfate on testicular lipid peroxide and GSH concentration as well as antioxidant enzymatic defense system. PMID- 18380168 TI - 13 Cis-retinoic acid mediates apoptosis in Dalton's lymphoma ascites cells by regulating gene expression. AB - Apoptosis is a form of regulated cell death that plays an important role in the maintenance of tissue homeostasis. Treatment of Dalton's lymphoma ascites cells (DLA)-induced experimental solid tumor tumor cells with 13 cis-retinoic acid, at concentrations of 25 microg/mL and 50 microg/mL, produced apoptotic morphologic changes, such as nuclear condensation and DNA-ladder formation after 48h incubation at 37 degrees C. in addition, the compound upregulated caspase-3 expression and downregulated bcl-2 gene expression. In vivo treatment with 13 cis retinoic acid (200 micromoles/kg body weight, ip) inhibited solid tumor development induced with DLA cells in Swiss albino mice. In conclusion, the results suggest that the anti-tumor activity of 13 cis-retinoic acid in DLA cells is due to apoptosis mediated by the regulation of bcl-2 and caspase-3 gene expression. PMID- 18380169 TI - Free cyanide-induced biochemical changes in nitrogen metabolism of the Indian major carp Cirrhinus mrigala. AB - We studied the effect of free cyanide on several aspects of nitrogen metabolism in the gills, liver, and muscle tissue of the Indian major carp, Cirrhinus mrigala. Free amino acid levels and protease activity were elevated following exposure to lethal and sublethal doses of free cyanide. At the lethal concentration, significant changes were noticed on day 4 for both parameters. The normal condition was restored only at the sublethal concentration. PMID- 18380170 TI - Amelioration of cisplatin induced nephrotoxicity in mice by an ethyl acetate extract of Lagerstroemia speciosa (L). AB - We studied an ethyl acetate (EtoAC) extract of leaves obtained from the medicinal and ornamental tree, Lagerstroemia speciosa L, for nephroprotective activity in cisplatin-induced acute renal injury in Balb/C mice. The EtoAC extract at dose levels of 50 and 250 mg/kg showed a dose-dependent reduction in cisplatin-induced elevations in urea and creatinine concentrations. Additionally, treatment with the EtoAC extract prevented the cisplatin-induced decline of the renal antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase, and reduced glutathione. The findings suggest that the EtoAC extract from L. speciosa possesses marked nephroprotective activity and could offer a promising role in the treatment of acute renal injury caused by a nephrotoxin like cisplatin. PMID- 18380171 TI - Adrenalectomy potentiates the anti-inflammatory activity of a calcium channel blocker. AB - Calcium movement is important in the activation of inflammatory cells. Prior studies have shown that calcium channel blockers (CCBs) inhibit carrageenan induced paw edema in rats by acting at pituitary and hypothalamic levels. Here we evaluated the role of calcium channel blockers and their regulation via the hypothalamus-pituitary-adrenal (HPA) axis in acute and chronic models of rat paw inflammation, using both carrageenan (acute) and formalin (chronic) as inflammation inducers. Adrenalectomized (ADX) and intact Sprague Dawley rats (n = 4 per group) weighing 150-250 each were treated with an intraperitoneal injection of nifedipine (400 microg/kg) or vehicle. Edema was assessed plethysmometrically by evaluating paw volume changes. The results show a significant contradiction between acute and chronic inflammation data. In intact animals, nifedipine demonstrated a significant anti-inflammatory effect in the acute inflammatory model but not in the chronic model. In adrenal-ectomized animals, the anti inflammatory effect of nifedipine was significantly enhanced both in acute and chronic models. We conclude that adrenalectomy plays a significant role in modulating the inflammatory pathway in the presence of calcium channel blockers. PMID- 18380172 TI - Hepatotoxic potential of Malathion in the freshwater teleost, Cirrhinus mrigala (Hamilton). AB - Freshwater edible fish, Cirrhinus mrigala, were exposed to a sublethal concentration (3 microl/L) of commercial grade malathion (50% EC) for 5, 15, or 25 days. After each exposure period, the liver was removed to study biochemical alterations. An increase in free amino acids, protease activity, and acetylcholine (Ach) levels, in contrast to decrement in total, structural, and soluble proteins, and acetylcholine esterase (AchE) activity were observed at 5 and 15 days of exposure, but on day 25 of exposure, all values reached near normalcy. The restoration to normalcy implies that after 15 d of exposure, an oscillatory phase in protein turnover toward a more-synthetic phase seems to occur, leading to the establishment of recuperation and adaptation phenomena. PMID- 18380173 TI - Arousal and continuous attention during Ramadan intermittent fasting. AB - During the month of Ramadan, practicing Muslims abstain from eating and drinking from sunrise to sunset. We aimed to investigate the effect of Ramadan fasting on arousal and continuous attention. The electrodermal activity and cancellation test of students were measured in fasting and non-fasting conditions after the conclusion of the Ramadan fast period. The skin conductance level of the fasting group was no different from the non-fasting group. In non-fasting group, the skin conductance response amplitude to an auditory stimulus was higher and the skin conductance response onset latency was lower than in the fasting group. Cancellation test results: the fasting group had a lower total number of marked targets (TNTM) but a higher total number of missed targets (TNMT) and length of time for the subject to complete the test (LTCT) than the non-fasting group. Ramadan fasting did not change arousal, but the reaction time to an auditory stimulus increased during the Ramadan intermittent fasting. Both reaction amplitude and continuous attention also decreased in the fasting condition. PMID- 18380174 TI - [Regulatory properties of Bacillus subtilis isolectins]. AB - The ability of natural and mutant Bacillus subtilis cultures with imperfect reparation/recombination system to synthesis of extracellular and surface lectins was investigated, and dependence of lectin production process on cultures' genotype was proved. Mutant B. subtilis recP has practically lost its ability to produce the extracellular lectins as a result of mutation of a gene of the reparation/recombination system. The application of the method "autofocusing" allowed to investigate all the spectrum oflectin molecular forms of natural B. subtilis culture and to reveal isoforms distinguished by physico-chemical and hemagglutination properties. It was shown that lectin cathode forms inhibit the transcription process from plasmid promnoter completely, and anodic forms activate the transcript formation slightly in the transcription in vitro with T7 bacteriophage DNA-dependent RNA-polymerase. PMID- 18380175 TI - [Chemical characteristic and serological activity of Pragia fontium lipopolysaccharides]. AB - Lipopolisaccharides (LPS) of 8 strains of Pragia fontium--representatives of a new genus of Enterobacteriaceae family, was isolated for the first time. It was demonstrated, that lipopolysaccharides yields were 10-20% of dry cells weight. Investigations of monosaccharide composition have determined the presence of glucose, galactose, rhamnose and mannose in lipopolysaccharides, that permitted to establish 4 chemotypes ofpolysaccharide part of LPS. Serological investigations showed immunochemical heterogeneity of Pfontium: 8 investigated strains were divided by the O-antigenicity into 7 serogroups. PMID- 18380176 TI - [Dynamics of successive changes in sulphidogenic microbial association under the conditions of formation of the biofilm on steel surface]. AB - Dynamics of successive changes in the artificial associative culture of sulphate reducing bacteria was investigated under biofilm formation on steel 3 specimens. It was shown that the ammonificating, denitrificating and iron-reducing bacteria were associative satellites of sulphate-reducing bacteria in the biofilm. Heterotrophic satellites ofsulphate-reductors with domination ofammonificating bacteria developed in the first hours of exposition in the microbial association. The appearance of sulphate-reducing bacteria in the biofilm was fixed on the 24th hour of exposition, their domination was noticed after 72 and 240 hours. It was shown that the successive changes with a consequent change of domination of different bacterial physiologic groups took place in the microbial association under the biofilm formation on the steel surface, that favoured mutually beneficial functioning of corrosion-dangerous bacteria. PMID- 18380177 TI - [Development of corrosion microbial association of soil in the presence of linuron and its derivatives]. AB - Qualitative stability of corrosion-dangerous microbial association, extracted from ferrosphere of corroding underground pipelines in different soils is shown. Prevailing of sulphate-reducing and iron-reducing bacteria is peculiar to the structure of formed corrosion-aggressive microbial cenosis. Development of association of iron-reducing bacteria in the presence of acting matter of pesticide Linuron and its derivatives is depressed. The association of sulphate reducing bacteria is sensitive only to the derivate that contains triazolazepine cycle and tolyl. PMID- 18380178 TI - [Sulfur-oxidizing bacteria of Yavoriv sulfur deposit reservoirs]. AB - Eleven pure bacteria cultures, able to oxidize thiosulfate during growth at pH 7.0-9.4, were isolated from surface layers of Yavoriv sulfur deposit open pit waters. Two cultures proved to be obligate aerobes, but nine cultures performed anaerobic respiration using nitrite, N2O (5 cultures) or only nitrite (4 cultures) as terminal electron acceptors. The growth of all cultures at 22 and 28 degrees C and growth absence at 35, 42 and 55 degrees C was established. All the bacteria are obligate chemolithoauthotrophs, because after cultivation with thiosulfate in the presence ofbiotin, yeast extract, formiate, succinate, arabinose, glucose, fructose and sucrose no growth stimulation was observed, heterothropic growth of any culture was not shown. As to their morphology the cells were bacillary, cytoplasmic membrane was surrounded by three-layer cell wall typical of gram negative bacteria intracellular inclusions, nucleoid, ribosomes and polysomes were also available. On the basis of obtained physiological and morphological characteristics the isolated bacteria cultures were referred to a group of neutrophiles, representatives of genus Thiobacillus, in particular to obligate chemolithoauthotrophs. PMID- 18380179 TI - [Generalized transduction of plasmid pKM101 by temperate bacteriophage ZF40 of Erwinia carotovora]. AB - It was shown that temperate bacteriophage ZF40 of Erwinia carotovora can perform generalized transduction of plasmid pKM 101. The antibiotic-resistance marker transfer is coordinated with the fact of cyclic permutation of the phage genom. The presented results create preconditions for further use of bacteriophage ZF40 as a convenient instrument for genetic study of E. carotovora. PMID- 18380180 TI - [Electron-microscopic study of potato aucuba mosaic virus]. AB - The results of electron-microscopic investigation of the peculiarities of virus interrelations with cells of mesophyl of the potato leaves, which were infected with potato aucuba mosaic virus (PAMV) are presented. Aggregates of the virus particles have been revealed which are localized in cytoplasm between the cellular organoids and just near the cellular membrane. The significant destructural changes of the structure of chloroplasts and formations of virus induced inclusions in them are observed. The cells contain cytoplasmic inclusions -round and oval electron-dence formations which are surrounded with a friable membrane. They occupy the large zones of cytoplasm, that is a typical sign of PAMV infection. PMID- 18380181 TI - [Microbe amylases: characteristic, properties and practical use]. AB - Current data concerning structure, properties and methods of purification ofmicrobial amylolytic enzymes are summarized in this paper. A short characteristic of the main methods of amylase activity measuring is presented, the advantages and disadvantages of each method are shown. It is proposed that novel techniques of enzyme immobilization stabilize the structure of amylases and allow their multiple uses. Scientific interest to amylases is analyzed that is explained by a number of their unique properties such as thermostability and pH tolerance. Authors have demonstrated some examples of the practical using ofamylases in different fields of industry: textile, paper, food industries, brewing and wine-making. The prospects of their possible using in detergent preparing for laundries and dishwashers are presented. It is supposed that future investigations in this trend for isolating new amrnylases from native producers will be developed. PMID- 18380182 TI - Prepare your revenue cycle for the future: a three-stage process. PMID- 18380183 TI - The U.S. mortgage crisis: its effect on healthcare debt. PMID- 18380184 TI - A good companion. PMID- 18380185 TI - Devising a system. New tools help therapists find seating solutions. PMID- 18380186 TI - Case challenge. Therapists combine resources to aid a child with spina bifida. PMID- 18380187 TI - Stay-at-home solutions for seniors. Home modification for aged and disabled on the rise. PMID- 18380188 TI - Consider the angles. Shoulder injuries require a 360-degree look at failure and prevention. PMID- 18380189 TI - Therapy with a charge. Iontophoresis offers therapists an effective way to address soft-tissue inflammation. PMID- 18380190 TI - Getting to the core. Using pilates to promote overall wellness during rehabilitation. PMID- 18380191 TI - Inpatient rehab overview. PMID- 18380192 TI - [Complex echography in the evaluation of the efficiency of treatment for some forms of lymphoproliferative diseases in the lesion of lymph nodes of the abdomen and retroperitoneal space]. AB - A complex ultrasound study of the abdominal and retroperitoneal lymph nodes afflicted by a lymphoproliferative process was made in 33 primary and 20 secondary (pretreated) patients prior to specific therapy and during treatment. The basic echographic signs of their involvement in the pathological process were as follows: higher dimensions, hypoechogenicity, and intranodal vascularization. The lymphoma-affected lymph nodes (in 10 patients) featured rich blood supply with color signals both in the center and around the periphery of a lymph node. The first changes in the diseased lymph nodes as a reduction in the number of intranodal vessels and/or vascular fragments were noted on echograms (in both primary and secondary patients) three days after the initiation of specific therapy if the latter was effective. The node became hypovascular or avascular; there was a rise in the echogenicity of abnormal lymph nodes. During continued antitumor treatment, echographic monitoring revealed regression of the involved lymph nodes or stabilization of the pathological process in 79% of the primary patients and in 80% of the secondary ones. In the remaining patients, the disease progressed in 21% of the primary patients and in 20% of the secondary ones. PMID- 18380193 TI - [Role of magnetic resonance imaging in the detection of chondral and osteochondral lesions in chronic juvenile arthritis]. AB - Chronic juvenile arthritis (CJA) is the most common inflammatory disease of joints in children. There are numerous studies showing the limited informative value of X-ray in the evaluation of CJA progression. Contrast-enhanced magnetic resonance imaging (MRI) using intravenous gadolinium is currently in the foreground in diagnosing arthritis in children, in infants in particular. Knee joints are most frequently afflicted in CJA, showing significant manifestations of the disease. The purpose of the study was to describe the patterns of changes in the nonossified epiphyseal and articular cartilages in the distal epiphyses of femurs in the knee joints of patients with manifestations of chronic juvenile arthritis and to define the role of contrast-enhanced MRI in evaluating the epiphyseal changes in this disease. Sixty-nine patients aged 1.5-14 years who have clinical laboratory and ultrasound signs of CJA lasting 6 months to 5 years underwent contrast-enhanced MRI for the evaluation of changes in the articular and nonossified epiphyseal cartilages. Intravenous contrast enhancement identified several specific features and patterns of epiphyseal changes: subchondral hyperemia of epiphyses and recorded thickened epiphyseal chondral vascular channels, chondral and osteochondral erosions as manifestations of changes in the growing epiphyseal cartilage and articular one in children with chronic arthritis. Thus, contrast-enhanced MRI allows differentiation of different patterns of epiphyseal changes in CJA. PMID- 18380194 TI - [Role of computed tomography in the estimation of the density of the bony block of an injured vertebral column segment at different stages of treatment with an external transpedicular fixation apparatus]. AB - A Somatom AR.HP computed tomograph (Siemens) was used to make studies in 54 patients with injury to the vertebral column and spinal cord in order to determine the formation of a bony block and its density in different portions of an injured segment at the stages of treatment. During therapy, the highest values of bony block density were observed in the dorsal portions of an injured segment (mean 377.32 +/- 184.57 HU). This may be accounted for by a slight effect of artefacts of an external transpedicular fixation apparatus, the main components of which are present at the posterior supporting complex. After treatment, the highest values of bony block density were noted in the central portions of an injured segment (mean 353.11 +/- 208.01 HU). This may be associated with rather serious intervertebral disk damage in this portion. The bony block density index was 339.75 +/- 139.41 HU in the left lateral portions and as high as 312.56 +/- 135.07 HU in the right ones. In the late period, the highest bony block density index (mean 397 +/- 193.07 HU) was seen in the right portions of an injured segment, which may be due to slight scoliotic deformity with the angle being open to the right. PMID- 18380195 TI - [Description of rearrangement of distraction reclaim in the extension of the lower extremities and elimination of their deformity in patients with various etiology of shortening]. AB - The data of X-ray and computed tomographic studies of a long tube distraction reclaim were analyzed in 80 patients who had limb shortening and deformity of varying etiology. The analysis has shown that during distraction there is a positive correlation between the increase in the connective-tissue interlayer area of a distraction reclaim and the overall increase in the area of the distraction reclaim (their ratio is not greater than 30% under the optimal elongation conditions). During fixation there is a decrease in the ratio of the area of a growth zone to the total area of a reclaim and in its total density with a parallel increase in the density of cortical laminae of a newly formed bone. Complete organotypic rearrangement comes to the end in the range of 1-3 years depending on the magnitude of pathomorphological and biomechanical changes and limb function. PMID- 18380196 TI - [Dynamic hepatobiliary scintigraphy in the evaluation of the efficiency of reconstructive surgery in patients after gastrectomy]. AB - The study was aimed at defining the informative value of dynamic hepatobiliary scintigraphy (HBSG) in the evaluation of duodenal reflux in patients with postgastrectomic disorders after reconstructive surgery. HBSG was performed in 23 patients. After intravenous injection of 99mTc bromeside in a dose of 74-80 MBq, the areas concerned were identified in the projection of the liver, gallbladder, hepaticocholedochus, duodenum, and stomach for 120 minutes with 60-min cholagogic breakfast. Histrograms were plotted in the activity-time coordinates; hepatic function, gallbladder contractility, and hepaticocholedochal patency were evaluated; the duodenal motor function index and the gastroduodenal reflux were estimated. HBSG is physiological, which makes it possible to reveal duodenal motor-and-evacuation disorders, to specify the reason for poor outcomes of gastrectomy, to objectively evaluate the severity of postgastrectomic disorders, and to determine further treatment policy for this difficult group of patients. PMID- 18380197 TI - [Changes in renal tumor contrasting in multiphasic SCT]. AB - Changes in renal tumor contrast and renal parenchyma-tumor density gradient were determined at multiphasic spiral computed tomography (SCT). A hundred and seven patients with renal tumors underwent SCT in the native phase and after bolus intravenous contrasting in the corticomedullary, nephrographic, and early excretory phases of the study. According to the level of contrast-enhancement in the corticomedullary phase, the neoplasms were divided into three groups: intensive, moderate, and low contrasting. Intensively and moderately contrast enhancement tumors showed the maximal contrast in the corticomedullary phase of the study with a significant reduction in density values in the nephrographic and early excretory phases. Lowly contrast-enhancement tumors generally demonstrated a slow gradual increase in their contrast from the native phase to the corticomedullary and nephrographic ones with their minimum contrast changes between the nephrographic and early excretory phases. In this group of patients, the difference was frequently insignificant between the neoplasmic density values in the native and corticomedullary phases in this group of patients, so it is expedient to use a combination of native and nephrographic or native and early excretory phases to make a differential diagnosis of hypovascular tumors and renal cysts. The maximum intact renal parenchyma-tumor density gradient was detectable in the nephrographic phase therefore this study phase is of the greatest information value in detecting renal tumors. PMID- 18380198 TI - [The role of low-field strength magnetic resonance imaging in bladder cancer staging]. AB - This article shows the role of magnetic resonance imaging (MRI) in complex diagnostics of urinary bladder cancer. The paper analyzes the authors' own data of urinary bladder MRI in 40 patients with histologically proven bladder cancer. This study demonstrates the additional capacities of low-field strength MRI with enhanced technique including conventional T1-, T2-weighted images along with FLAIR and PD images. PMID- 18380199 TI - [Radiodiagnosis in spine injuries]. PMID- 18380200 TI - [Radiodiagnosis of injuries to the eye and orbital structures]. PMID- 18380201 TI - Increase in collagen turnover induced by intradermal injection of carbon dioxide in rats. AB - INTRODUCTION: Results from clinical observations have demonstrated that percutaneous infiltration of carbon dioxide improves the appearance of the skin in adjacent areas. No studies have been found in the literature that showed evidence of histological changes caused by carbon dioxide injections. OBJECTIVES AND METHODS: A blind cross-sectional pilot study was performed in the Departments of Pharmacology and Morphology of the Federal University of Ceara, with the aim of histologically investigating whether intradermal and/or subcutaneous injection of medicinal carbon dioxide would increase collagen turnover in rats. Ten male Wistar rats were used, aged 3 months (2 animals) and 14 months (8 animals). The 2 younger rats were used as controls. Four of the older rats received injections of saline solution (0.9%), and were also considered to be controls. In the remaining 4, carbon dioxide was injected into the subcutaneous cellular tissue and intradermally. Biopsy samples were collected before and after treatment with carbon dioxide. RESULTS: Collagen turnover increased in the treated animals in comparison with the controls. Compression of collagen bundles in the tissue samples where intradermal injection was used was more intense than in the subcutaneous treatment. The histological characteristics of the samples with carbon dioxide injected intradermally were similar to the characteristics of the younger rats (controls). CONCLUSIONS: The results obtained corroborate clinical observations of aesthetic improvements in the facial skin with carbon dioxide injections. Future research should address the comparison between intradermal and subcutaneous injections, the volume of gas used, and the frequency of treatment sessions. PMID- 18380202 TI - Dermatological implications of skeletal aging: a focus on supraperiosteal volumization for perioral rejuvenation. AB - It is becoming widely accepted that volume changes in the skin and soft tissue contribute greatly to age-related facial reshaping. A significant contribution to these volume changes is the loss of craniofacial skeletal support to the overlying soft tissue. Gravity, once considered the major culprit in facial aging, is now recognized to determine the direction, rather than the extent, of tissue deflation. Although the sequence of events observed in aging is somewhat predictable, its pace among individuals is variable and may be influenced by both intrinsic (e.g., gender, genetics) and extrinsic (e.g., photoaging, smoking, stress) factors. Changes in different tissue layers within a single individual do not occur independently, but interdependently; changes in one tissue within an individual may influence subsequent changes in other tissues. Midfacial soft tissue descent has been observed in response to decreased craniofacial support in both congenital craniofacial hypoplasia and following trauma, leading to a hypothesis that the loss of underlying bony support for any reason, including aging, leads to soft tissue descent in the face. As craniofacial support (the "table") decreases, it leaves less surface area for the outer soft tissue envelope (the "tablecloth") causing it to fold or sag. Replacing this deep support with craniofacial implants has been shown to reposition the overlying soft tissue. Following a brief review of the current literature on aging changes in the skin, soft tissue, and bone; the authors describe their experience with the use of poly-L-lactic acid (PLLA), both as a soft tissue volumizer and as an injectable craniofacial implant in a supraperiosteal location to address both soft tissue volume loss and loss of craniofacial support. In the cases presented, the most striking result noted was the ability to restore a youthful proportion to the perioral area, which had not been achieved previously with soft tissue treatment alone. PMID- 18380203 TI - Botulinum toxin type a versus topical 20% aluminum chloride for the treatment of moderate to severe primary focal axillary hyperhidrosis. AB - Severe hyperhidrosis affects 2.8% of the population and can be emotionally devastating. First-line therapy employs topical agents such as aluminum chloride (AC), but efficacy and tolerability vary widely. Botulinum toxin type A (BTX-A) is FDA-approved for the treatment of primary focal axillary hyperhidrosis unresponsive to topical therapy. A single-center, randomized, parallel, open label, 12-week study was performed to compare the efficacy and safety of BTX-A with 20% AC for the treatment of primary focal axillary hyperhidrosis. Twenty five subjects were randomized to either BTX-A or AC treatment, and were evaluated for treatment response by an improvement of > or =2 grades on the Hyperhidrosis Disease Severity Scale (HDSS). At week 4, 92% of the subjects in the BTX-A group achieved treatment response compared with 33% of the subjects in the AC group. Overall, treatment with BTX-A was more effective and provided greater patient satisfaction than with AC. Treatment with AC was effective and tolerated in 29% of the subjects. PMID- 18380204 TI - Histological and clinical studies on the effects of low to medium level infrared light therapy on human and mouse skin. AB - BACKGROUND AND OBJECTIVE: Deep heating or denaturation of collagen has been reported to be necessary for nonablative skin rejuvenation. The purpose of this study was to examine whether thermally damaged collagen is an indispensable factor to increase the amount of collagen in vivo. Epidermal and dermal responses to infrared light therapy using a Titan source were examined with the aim of correlating histological and clinical responses in human and amelanotic mouse skin. STUDY DESIGN/MATERIALS AND METHODS: Ten, 20, or 30 J/cm2 infrared light were irradiated on the human subject's skin (thigh), while 5, 10, 20, or 30 J/cm2 were used on amelanotic mouse skin. Biopsies were taken and analyzed using hematoxylin and eosin (H&E) and Elastica von Gieson stain. RESULTS: Ten or 20 J/cm2 infrared light increased the amount of both collagen and elastin in all layers of the dermis without denaturing the collagen in human skin. A higher dose of 30 J/cm2 also increased the amount of collagen and elastin, but denatured the collagen in human skin. (In addition to the thigh, 2 treatments of 10 J/cm2 infrared light improved skin toning and texture on the subject's face). In mouse skin, 5 or 10 J/cm2 remarkably increased the amount of both collagen and elastin, and of epidermal cells. Twenty or 30 J/cm increased the amount of collagen and elastin and the number of keratinocytes, but caused some vacuolated degeneration of keratinocytes. The presence of denatured collagen was not evident due to the high density of collagen. CONCLUSIONS: This study shows that the denaturation of collagen is not required to increase the amounts of collagen or elastin in vivo in human skin. The activation of the mitochondria as well as the denaturation of collagen may play important roles in infrared phototherapy. PMID- 18380205 TI - Lip augmentation and rejuvenation using a novel, porcine collagen-derived filler. AB - Proportionally projecting feminine lips are considered attractive sexually. With aging, predictable changes occur in the lips, which often prompt women to seek lip rejuvenation for enhancement. Although there is an ever-expanding list of products for lip augmentation, very little technical information and guidelines for correct enhancement are available in the dermatological literature. This review provides clinical guidelines for a systematic approach to lip augmentation and rejuvenation, and documents clinical experience with Evolence Breeze, a novel, porcine collagen-derived dermal filler. Before undertaking such procedures, thorough lip assessments and clearly defined treatment objectives are needed. Fifteen women, ages 42 to 61 years, underwent procedures using Evolence Breeze; 6 required augmentation and 9 required rejuvenation. Only minimal swelling was noted immediately after lip injection. At 3 months, results were rated as "very good" by 8 patients (53%), "good" by 5 (33%), and "satisfactory" by 2 (13%). At 6 months, all had ongoing results. PMID- 18380206 TI - Utilization of narrow-band ultraviolet light B therapy and etanercept for the treatment of psoriasis (UNITE): efficacy, safety, and patient-reported outcomes. AB - BACKGROUND: Moderate to severe psoriasis is a significant inflammatory disease that frequently requires systemic therapies to effectively treat the underlying disorder. Etanercept and narrow-band ultraviolet light B (NB-UVB) are widely used to treat this disease. OBJECTIVE: To evaluate the effectiveness, tolerability, and patient-reported outcomes of combination etanercept plus NB-UVB phototherapy in moderate to severe plaque psoriasis. METHODS: This 12-week, single-arm, open label study evaluated the combination of etanercept 50 mg twice weekly and NB-UVB thrice weekly in 86 patients. The primary outcome measure was > or =75% improvement from baseline in the Psoriasis Area and Severity Index (PASI 75). Other measures included PASI 90, PASI 100, and the Dermatology Life Quality Index (DLQI). RESULTS: At week 12, 26.0% achieved PASI 100, 58.1% achieved PASI 90, and 84.9% of patients achieved PASI 75. Mean improvement from baseline in DLQI was 84.4%. No unexpected, untoward adverse events were noted. CONCLUSIONS: A 12-week course of etanercept plus NB-UVB phototherapy was well tolerated and produced clinically meaningful improvements in signs and symptoms of moderate to severe plaque psoriasis and in patient-reported outcomes. Further investigation of the safety and efficacy of the use of such combination for this indication in controlled clinical trials would be of interest. PMID- 18380207 TI - A pilot study to determine the safety and efficacy of monochromatic excimer light in the treatment of vitiligo. AB - BACKGROUND AND OBJECTIVE: According to a European pilot study, the 308-nanometer (nm) Excilite mu (DEKA, Florence, Italy) system may be a promising tool for patients with vitiligo by offering targeted phototherapy, a rapid onset of repigmentation, and few adverse effects. The objective of this study was to evaluate the clinical efficacy and safety of the 308-nm Excilite mu in the treatment of vitiligo. METHODS AND LIMITATIONS: Ten patients with stable vitiligo were exposed to 10 weeks of targeted phototherapy with the Excilite mu device, followed by 5 weeks of observation. Skin types 1 and 2 were not included in the cohort, and Wood's light examination was not documented. RESULTS: At 2 weeks, repigmentation was observed in 60% of the subjects, according to patient assessment, and 50% of the subjects, according to the treating physician and independent observer assessments. All patients maintained the repigmentation during the 5-week, follow-up period. CONCLUSION: The 308-nm Excilite mu is a safe and fast-acting therapeutic option in patients with stable vitiligo and skin types 3 through 6. PMID- 18380208 TI - Hydrocortisone butyrate 0.1% lotion in the treatment of atopic dermatitis in pediatric subjects. AB - BACKGROUND: Hydrocortisone butyrate (HCB) is currently marketed as a cream, ointment, and solution. A new lotion formulation of hydrocortisone butyrate 0.1% (Locoid lotion) has been developed and evaluated. OBJECTIVE: The objective of this study was to evaluate the efficacy and safety of HCB 0.1% lotion compared to the vehicle in subjects aged 3 months to less than 18 years with mild to moderate atopic dermatitis (AD). METHODS: In this multicenter double-blind study, 284 subjects with mild to moderate AD were randomized 1:1 to receive HCB 0.1% lotion or the vehicle for a duration of 4 weeks. "Treatment success" was defined as those subjects with a final Physician Global Assessment (PGA) score of 0 or 1 that had at least a 2-point reduction in the PGA score from baseline to day 29. Safety was assessed by monitoring adverse events. RESULTS: Analyses of the final PGA score showed a significant treatment effect (P <.001) in favor of the HCB 0.1% lotion group. The safety profile of the HCB 0.1% lotion was also favorable. LIMITATIONS: The study did not assess the durability of the treatment effects (ie, safety and efficacy) after completion of the 4-week treatment period nor the potential need for longer term therapy given the chronic nature of AD. CONCLUSION: Results demonstrate the safety and efficacy of HCB 0.1% lotion in the treatment of mild to moderate AD in children 3 months to 18 years of age. PMID- 18380209 TI - Skin rejuvenation in Asian skin: the analysis of clinical effects and basic mechanisms of intense pulsed light. AB - BACKGROUND: Skin aging consists of photoaging and intrinsic aging. It is characterized clinically not only by rhytides, but also by pigmentary alterations and facial telangiectasias. There continues to be a growing interest in the efficacy of intense pulsed light (IPL) devices in the treatment of skin aging, as well as further defining its mechanism of action. OBJECTIVES: The objective of this clinical trial was to evaluate the effects and the mechanism of action of an IPL by comparing clinical photographs and biopsy results before and after treatment. METHODS: A total of 58 patients were treated using a new IPL device. Clinical photographs were taken before treatment and compared to those taken 3 weeks after the treatment. Also, 4 cases had pathological analyses of tissues that were stained by haematoxylin-eosin and Uana orcein. Immunohistology of human collagen of types 1 and 3 and quantitative analyses of elastin and collagen were performed by a poly-functional digital image light microscope; a transmission electron microscope was used for 2 of the cases to look for additional changes. RESULTS: After 3 treatments, 62.1% of the patients showed improvement in wrinkles and skin texture. Pigmentation improved in 84.6% of the patients, and a reduction in telangiectasis was seen in 81.25% of the patients. Pathological examination showed that both type 1 and type 3 collagens increased following treatment, but elastin content decreased; however, the elastin fibers were arranged more neatly. In the transmission electron microscope study, the amount of fibroblast activity increased, the fibroblasts were more active, and there were more collagen fibers neatly rearranged within the stroma. CONCLUSION: Clinical and pathological studies demonstrated that the IPL was effective in improving wrinkles and skin texture. The mechanism of action may be through the increasing activity of the fibroblasts, hyperplasia of the fibroblasts, and rearrangement of both collagen and elastin within the stroma. PMID- 18380210 TI - Fast and easy treatment for reduction of the Tyndall effect secondary to cosmetic use of hyaluronic acid. AB - The use of hyaluronic acid (HA) filler substances for facial skin wrinkling and volume loss has become popular. With increasing use, physicians are likely to encounter side effects. One such side effect is a bluish discoloration known as the Tyndall effect. Recently, we have encountered a few cases in which patients presented with a bluish tint in the areas where hyaluronic acid was injected. A number of treatments can be performed to reverse these unwanted effects. We present a simple method for correction of misplacement of HA gel. PMID- 18380211 TI - Erythromelalgia of the ears: an unusual variant and response to therapy. AB - Erythromelalgia is characterized by episodes of erythematous, warm, burning acral skin, which is exacerbated by heat and relieved by cold. Erythromelalgia usually affects the feet and/or hands but, although rare, erythromelalgia may affect the ears. The authors present a 65-year-old woman with erythromelalgia of the ears with disabling symptoms whose diagnosis was delayed for 6 years. The patient failed to respond to numerous therapies before rapidly improving with oral amitriptyline and amitriptyline 1% to 2% and ketamine 0.5% to 1% topical gel. PMID- 18380212 TI - Basal cell carcinoma of the vulva presenting as unilateral pruritus. AB - Basal cell carcinoma (BCC) is the most common human malignancy, which often occurs as a result of ultraviolet light on sun-exposed areas. A more rare location for the presentation of BCC is the non-sun-exposed genital area, where squamous cell cancer occurs frequently in the setting of human papilloma virus and chronic inflammatory lesions (i.e., lichen sclerosus et atrophicus). Consequently, such tumors may escape detection by the dermatologist and be mistaken by the gynecologist for an inflammatory condition. A delay in diagnosis can result in wider surgical margins and potential recurrences. We present a case of BCC of the vulva with involvement of the clitoris presenting with unilateral pruritus and treated as an allergic contact dermatitis with topical corticosteroids. The patient was treated with Mohs micrographic surgery in conjunction with topical imiquimod to spare surrounding tissue. PMID- 18380213 TI - Inguinal hyperhidrosis misdiagnosed as urinary incontinence: treatment with botulinum toxin A. AB - Hyperhidrosis is a psychosocially embarrassing condition with a treatment history of varied success, including topical application of antiperspirants with aluminum salts (e.g., Drysol), oral anticholinergics, iontophoresis, and endoscopic transthoracic sympathectomy. The use of botulinum toxin type A (Botox) in humans for the treatment of hyperhidrosis didn't emerge until 1996. Botulinum toxin A is a safe and effective treatment for hyperhidrosis, and it has been shown to improve the quality of life in affected patients. The toxin works by inhibiting the release of acetylcholine at the neuromuscular junction, and affecting the postganglionic sympathetic innervation of sweat glands. The authors report a case of inguinal hyperhidrosis treated with botulinum toxin A. Very few cases with similar treatments have been found in the medical literature. More work should be done to find the optimal dose for treating this area, and affected patients should be informed of the potential benefits of botulinum toxin therapy. PMID- 18380214 TI - Somatic stem cells in dermatology. PMID- 18380215 TI - Cutaneous clues to drug addiction. PMID- 18380216 TI - Herpesvirus infection in European flat oysters Ostrea edulis obtained from brood stocks of various geographic origins and grown in Galicia (NW Spain). AB - We evaluated differences in productive traits and disease susceptibility among Ostrea edulis stocks. We produced 4 to 5 families from each of 4 oyster populations (Irish, Greek and 2 Galician) in a hatchery. Spat corresponding to 19 different families were transferred to a raft in the Ria de Arousa (Galicia, Spain) for grow-out. Samples of each family were histologically processed every month for 2 yr. One of the pathological conditions disclosed by histological examination was characterised by the occurrence of numerous abnormal cells throughout the connective tissue of various organs, showing hypertrophied nuclei with marginated chromatin and a characteristic large intranuclear acidophilic inclusion. Ultrastructural examination showed that the abnormal cells contained herpesvirus-like particles. In situ hybridisation assay using a DNA probe specific for Ostreid herpesvirus 1 (OsHV-1) confirmed that the abnormal cells were infected by OsHV-1 or a closely related herpesvirus. All cases of this pathological condition, except one, were detected during the first year of grow out; thus it was mostly restricted to juvenile stages. The disease was detected in oysters of each origin but it was not found in all families of each origin, thus suggesting significant parental influence in the susceptibility to this disease or significant influence of the infective status of the parents on the infection of the progeny (vertical transmission). This pathological condition was likely responsible for oyster mortality to some extent during the first year of grow-out. PMID- 18380217 TI - Genotyping of white spot syndrome virus prevalent in shrimp farms of India. AB - DNA extracts from white spot syndrome virus (WSSV) that had infected post-larvae and juveniles of cultured shrimp, wild shrimp and crabs, which had been collected from different hatcheries and farms located along both the east and west coasts of India, revealed considerable variation in several previously identified WSSV DNA repeat regions. These include the 54 bp repeat in ORF 94, the 69 bp repeat in ORF 125 and the compound 45 and 57 bp repeat region in ORF 75. In ORF 94, 13 genotypes were observed with the number of repeats ranging from 2 to 16 units. While 7 repeat units were commonly observed (11.3%), no samples with 11 or 15 repeat units were found. In ORF 125, 11 types were found, with repeats ranging from 2 to 14 units. The most prevalent genotype displayed 4 repeat units (47.1%); no samples with 6 or 13 repeats were observed. The compound repeat region of ORF 75 displayed 6 different patterns of repeats. Samples with the same repeat pattern in one ORF did not always show identical repeat patterns in one or both of the other repeat regions. These data suggest that combined analysis of all 3 variable loci could be used to differentiate and characterize specific WSSV strains. For general epidemiological studies, the best marker with maximum variation is ORF 94, followed by ORF 125 and ORF 75. The 3 repeat regions above were used to compare WSSV genotypes from disease outbreaks on 3 sets of farms from different locations in the state of Andhra Pradesh. The genotypes within each farm set were almost identical, but differed between farm sets, suggesting that WSSV transmission occurred directly through virus carriers or water exchange between adjacent farms at each location. These findings show that genotyping can be a useful epidemiological tool for tracing the movement of WSSV within infected populations. PMID- 18380218 TI - Risk of introducing viral hemorrhagic septicemia virus (VHSV) to the Chilean South Pacific via sardine imports from Europe. AB - Chile imports from Spain 100s of metric tons of frozen sardine Sardina pilchardus fished in European oceans, which, with several other clupeids, are presumed susceptible to infection with viral hemorrhagic septicemia virus (VHSV). The frozen sardines are directly introduced into the sea as bait to catch southern hake Merluccius australis in the same areas where wild and pen-raised salmonids are present. A simulation model was therefore developed to evaluate the potential risk of infection of wild Chilean southern hake with VHSV from imported bait. The model indicated that VHSV-susceptible fish species present in Chilean waters, like southern hake, are not at immediate risk of infection. However, sensitivity analyses showed that infectious doses at lower concentrations of VHSV combined with higher VHSV-prevalence import scenarios could likely result in VHSV infections of a moderate number of indigenous southern hake (> or =54 fish yr( 1)). PMID- 18380219 TI - Outbreaks and risks of infectious spleen and kidney necrosis virus disease in freshwater ornamental fishes. AB - We examined the distribution of iridoviruses in 10 freshwater ornamental fish species hatched in Korea and imported from other Asian countries using both 1 step and 2-step polymerase chain reation (PCR). None of the 10 fish species analyzed were free of iridovirus as shown by 2-step PCR positive results, and 3 species yielded 1-step PCR positive results with associated mortality. Cloned PCR amplicons of the adenosine triphosphatase (ATPase) and major capsid protein (MCP) genes in genomic DNA of iridovirus showed the same nucleotide sequences as that of infectious spleen and kidney necrosis virus (ISKNV) isolated from the mandarinfish Siniperca chuatsi. These results indicate the presence of ISKNV disease in various ornamental fish as new host species and that the disease is widespread throughout different Asian countries including Korea, Singapore and China. Such infections were either clinical with associated mortality (and 1-step PCR positive) or asymptomatic in fish that were externally healthy (and only positive in 2-step PCR). Molecular analyses of the K2 region performed on iridovirus samples isolated from freshwater ornamental fishes revealed deletion/insertion of repetitive sequences of various lengths (42 to 339 bp), depending on the ISKNV isolates, without substitutions. Experimental infection of pearl gourami Trichogaster leeri and silver gourami T. microlepis with a tissue homogenate of pearl gourami infected by ISKNV induced 70 and 20% cumulative mortalities in the pearl and silver gourami, respectively. PMID- 18380220 TI - Neoparamoeba perurans is a cosmopolitan aetiological agent of amoebic gill disease. AB - Previously we described a new member of the Neoparamoeba genus, N. perurans, and showed that it is an agent of amoebic gill disease (AGD) of Atlantic salmon Salmo salar cultured in southeast Tasmania, Australia. Given the broad distribution of cases of AGD, we were interested in extending our studies to epizootics in farmed fish from other sites around the world. Oligonucleotide probes that hybridise with the 18S rRNA of N. perurans, N. branchiphila or N. pemaquidensis were used to examine archival samples of AGD in Tasmania as well as samples obtained from 4 host fish species cultured across 6 countries. In archival samples, N. perurans was the only detectable amoeba, confirming that it has been the predominant aetiological agent of AGD in Tasmania since epizootics were first reported. N. perurans was also the exclusive agent of AGD in 4 host species across 6 countries. Together, these data show that N. perurans is a cosmopolitan agent of AGD and, therefore, of significance to the global mariculture industry. PMID- 18380221 TI - Metabolism of praziquantel in kingfish Seriola lalandi. AB - Investigations into the metabolism of drugs used in aquatic animal therapy are useful for understanding the mechanisms of xenobiotic transformation systems and can aid the development of dosing regimens. This study investigated the metabolism of the synthetic anthelmintic praziquantel, which has application in helminthiasis treatment for several fish species including kingfish Seriola lalandi, a commercial aquaculture finfish species. At least 7 mono- or dihydroxylated derivatives of the parent compound were identified in kingfish after administration of a 150 mg kg(-1) oral praziquantel dose, paralleling findings in mammals. The structure of one representative mono-hydroxylated species that was prominent in the skin, muscle, liver, kidney and plasma of kingfish was investigated using fragmentation experiments; this revealed that hydroxylation of the parent molecule occurred in the tetrahydroisoquinoline region of praziquantel, analogous with mammalian metabolites, but different to that of the active mammalian metabolite (trans-4-OH-praziquantel). The implications of these findings with regard to biotransformation systems for this drug in mammals and fish are discussed. PMID- 18380222 TI - Hypoxia increases intensity of epidermal papillomatosis in roach Rutilus rutilus. AB - Hypoxia, which occurs frequently in aquatic ecosystems and is mainly due to increasing eutrophication can cause severe environmental stress in fish. We investigated experimentally the hypothesis that hypoxia could be one of the environmental stress factors that can induce papillomatosis in fish. Male roach Rutilus rutilus exposed to periodic oxygen deficiency and accompanied temperature increases (OT group) showed the highest increase in the intensity of papillomatosis, as measured by the number of scales covered by papillomatosis tumors. The second highest increase in disease intensity was among male roach exposed to periodical temperature increases. The incidence of such tumors was lowest in the control group, which was exposed to neither hypoxia nor increased temperature. The mortality of fish during the 17 d experiment was highest and the condition factor was lowest in the OT group, indicating this group experienced a higher level of stress. The apparent interaction of hypoxia and temperature suggests that these environmental stressors are among the multifactorial elements leading to papillomatosis in roach. Furthermore, these results provide experimental evidence to indicate that hypoxia may contribute to tumor development in fish. PMID- 18380223 TI - Perkinsus chesapeaki in stout razor clams Tagelus plebeius from Delaware Bay. AB - Perkinsus chesapeaki is reported from stout razor clams Tagelus plebeius in Delaware Bay, extending the known range of P. chesapeaki north of Chesapeake Bay. P. marinus, which causes dermo disease, is prevalent in cultured and wild oysters at this site, but was not detected in T. plebeius. Evidence for the presence of disseminated neoplasia, also reported from Chesapeake Bay, was equivocal. Although P. chesapeaki infections were associated with mortality events, light infection intensities and a general lack of histopathological evidence of disease limit inferences about a causal relationship. A comparison of Ray's fluid thioglycollate medium (RFTM)-based and PCR-based detection assays highlight differences in detection capabilities related to the quantity and type of tissue processed rather than assay sensitivity per se, a point that should be considered when surveying populations for disease prevalence. Investigators are further cautioned to use care when applying and interpreting diagnostic assays when used with novel species. PMID- 18380224 TI - Short- and long-term dietary effects on disease and mortality in American lobster Homarus americanus. AB - The American lobster Homarus americanus fishery is heavily dependent on the use of fish as bait to entice lobsters into traps. There is concern that this food supplementation is nutritionally insufficient for lobsters, but previous experiments reported conflicting results. We conducted a long-term feeding experiment in which 1 yr old American lobsters were fed one of 7 diets for a period of 352 d, a time that allowed the lobsters to molt thrice. The diets consisted of fresh frozen herring, a 'wild' diet (rock crab, mussel, and Spirulina algae), a formulated artificial diet for shrimp, paired combinations of these 3 diets or a diet formulated at the New England Aquarium (Artemia, fish and krill meal, Spirulina algae, soy lecithin, vitamins and minerals). The lobsters fed the diet of 100% fish had higher initial molting rates, but within the period of this experiment all either contracted shell disease or died. Mixed diets resulted in higher survival and a lower probability of mortality. This research demonstrated a critical time component to diet studies in lobsters. Short- and long-term impacts of diet differ. In the long term, continual high consumption rates of fish by the lobsters promote poor health in all lobsters, not just those of market size. The use of fish as bait may make lobsters more susceptible to the stress associated with environmental fluctuation, thereby leading to increased disease and mortality. This nutritional stress can be used to develop a laboratory model of shell disease in American lobsters. PMID- 18380225 TI - Distribution and morphology of growth anomalies in Acropora from the Indo Pacific. AB - We assessed the distribution and prevalence of growth anomalies (GAs) in Acropora from French Frigate Shoals (Hawaii, USA), Johnston Atoll and Tutuila (American Samoa), developed a nomenclature for gross morphology, characterized GAs at the cellular level and obtained preliminary indices of their spatial patterns and progression within coral colonies. Acropora GAs were found in all 3 regions, but the distribution, variety and prevalence of Acropora GAs was highest in American Samoa. GAs were grouped into 7 gross morphologies (exophytic, bosselated, crateriform, nodular, vermiform, fimbriate or annular). On histology, GAs consisted of hyperplastic basal body wall (calicodermis, mesoglea and gastrodermis apposed to skeleton) with 3 distinct patterns of necrosis. There was no evidence of anaplasia or mitotic figures (common but not necessarily required morphologic indicators of neoplasia). Compared to normal tissues, GAs had significantly fewer polyps, zooxanthellae within the gastrodermis of the coenenchyme, mesenterial filaments and gonads but significantly more necrosis. On 2 colonies with GAs monitored at 2 points over 11 mo, numbers of GAs per colony increased from 0.9 to 3 times the original number seen, and significant clustering of GAs occurred within colonies. The evidence of GAs being true neoplasias (tumors) is mixed, so a cautionary approach is urged in use of morphologic terminology. PMID- 18380226 TI - Dioxin--contemporary and future challenges of historical legacies. Dedicated to Prof. Dr. Otto Hutzinger, the founder of the DIOXIN Conference Series. PMID- 18380227 TI - Literature on dioxins and other POPs. PMID- 18380228 TI - 2nd Agent Orange and Dioxin Remediation Workshop. Hanoi, Viet Nam, 18-20 June 2007. PMID- 18380229 TI - Conjoint analysis: a useful tool for assessing preferences for environmental issues. PMID- 18380230 TI - Immobilization of lead in shooting range soils by means of cement, quicklime, and phosphate amendments. AB - BACKGROUND, AIM, AND SCOPE: Lead (Pb) contamination at shooting range sites is increasingly under environmental concern. Controlling Pb leachability from shooting range soil media is an important step to minimize Pb exposure to the surrounding environment. This study investigated stabilization of Pb in shooting range soils treated with cement, quicklime, and phosphate. MATERIALS AND METHODS: Two soils were used and collected from two shooting ranges, referred to as SR1 and SR2. The treatment additives were applied to the soils at rates from 2.5% to 10% (w/w). The effectiveness of each treatment was evaluated by Pb (w/w). The effectiveness of each treatment was evaluated by Pb leachability, measured by the Toxicity Characteristic Leaching Procedure (TCLP). The possible mechanisms for Pb immobilization were elucidated using X-ray powder diffraction (XRPD). RESULTS: Cement and quicklime treatments were effective in immobilizing Pb in SR1 soil, with reduction of Pb concentration in TCLP leachate (TCLP-Pb) to be below the U.S. EPA non-hazardous regulatory limit of 5 mg L(-1) at application rates of > or =5% and 28-d incubation. By contrast, cement and quicklime amendments were less effective for Pb stabilization in SR2 soil because the TCLP-Pb levels in the treated soil were still higher than the limit of 5 mg L(-1) at all application rates, although they were significantly reduced in comparison with the untreated soil. Phosphate application was most effective in reducing Pb leach ing in both soils. Even at an application rate as low as 5% and 1-d incubation, phosphate could reduce TCLP-Pb to be below the limit of 5 mg L(-1) in both soils. DISCUSSION: Immobilization of Pb in the SR1 soil amended with cement and quicklime was attributed to the formation of pozzolanic minerals (e.g., calcium silicate hydrate C-S-H and ettringite) that could encapsulate soil Pb. The pozzolanic reaction was limited in the SR2 soil upon the application of cement and quicklime. Reduction of the TCLP-Pb might result from complexation of Pb on the surface of the formed calcite. Phosphate-induced Pb immobilization was mainly attributed to formation of less soluble PbHPO4. CONCLUSIONS: The results indicate that effectiveness of cementitious treatments (cement and quicklime) in immobilizing Pb varies in two soils, being effective in SR1 soil but less in SR2 soil. For one given soil, no difference was observed of the effeciveness between cement and quicklime treatments, whereas phosphate amendment emerges as a most effective treatment means for stabilizing Pb in both two soils, and it also shows a faster immobilization process and little effect on the soil acid buffering capacity. Recommendations and Perspectives. Overall, our study reveals that immobilizing Pb can be one of the best management practices for Pb contamination at shooting range sites. Phosphate amendment is most effective in immobilizing Pb in any kind of the soil ranges to minimize negative Pb impacts on the shooting range sites. PMID- 18380232 TI - Master's Degree in sustainable development in Switzerland, the first master course comprising three faculties. AB - Sustainable development has become a key aspect in society, economics and environment. Therefore, experts dealing with questions relating to people, the environment and its resources are more and more requested. This paper presents the concept and first experiences of a specialised Master's Degree in Sustainable Development (MSD). This is a pioneer course as it is equally anchored in three faculties (Human science, Natural science, Business and Economy) at the University of Basel, Switzerland. It aims to transmit knowledge, teach methodology and enable practical work experience in the field of sustainable development. This interdisciplinary master's degree is composed of several modules. At first, the attendance of modules providing a basic understanding in the disciplines not yet covered by the former Bachelor degree, is mandatory. In optional modules, the acquired knowledge of the compulsory modules is further enhanced, focussing on four different topics and are titled as: Agglomeration and Ecosystems; Conservation and Utilisation of Natural Resources; Environment, Values, Societal Transformation and Health; and Environmental Problems in a Globalised World. In another optional module, students may complete an internship in which they can apply theoretical and thematic knowledge. To work independently on a problem in the context of, interdisciplinary projects are a central request in the MSD. Finally, the master thesis has to be planned and realized by a plying the scientific methods and skills acquired in the previous modules. Since the beginning of the programme in the winter of 2005/2006, 45 students have enrolled. They received degrees at 25 different universities, 13 of which are from abroad. Some already have several years of working experience, while others have only just completed their Bachelor's degrees. A analysis has shown that the graduates will have excellent chances in the employment market, since they are well qualified to take over sought-after positions in the business world, in administration, in consultancy, national or international organisations, or to follow academic careers. PMID- 18380231 TI - Humic substances. Part 2: Interactions with organisms. AB - GOAL, SCOPE AND BACKGROUND: Freshwater bodies which chemistry is dominated by dissolved humic substances (HS) seem to be the major type on Earth, due to huge non-calcareous geological formations in the Northern Hemisphere and in the tropics. Based on the paradigm of the inertness of being organic, direct interactions of dissolved HS with freshwater organisms are mostly neglected. However, dissolved organic carbon, the majority of which being HS, are natural environmental chemicals and should therefore directly interact with organisms. Major results that widened our perspective on humic substance ecology come from experiments with the compost nematode, Caenorhabditis elegans, which behaved contradictorily to textbook knowledge and provoked an in-depth re-consideration of some paradigms. APPROACH: To overcome old paradigms on HS and their potential interactions with organisms, we reviewed recent international literature, as well as 'grey' literature. We also include results from own ongoing studies. RESULTS: This review focuses on direct interactions of dissolved HS with freshwater organisms and disregards indirect effects, such as under-water light quenching. Instead we show with some macrophyte and algal species that HS adversely interfere with photosynthesis and growth, whereby closely related algal species show different response patterns. In addition to this, HS suppress cyanobacteria more than eukaryotic algae. Quinones in the HS appear to be the effective structure. Furthermore, HS can modulate the offspring numbers in the nematode C. elegans and cause feminization of fish and amphibians--they possess hormone-like properties. The ecological consequences of this potential remain obscure at present. HS also have the potential to act as chemical attractants as shown with C. elegans and exert a mild chemical stress upon aquatic organisms in many ways: induction of molecular chaperons (stress proteins), induction and modulation of biotransformation and anti-oxidant enzymes. Furthermore, they produce an oxidative stress with lipidperoxidation as one clear symptom or even stress defense strategy. Stronger chemical stresses by HS may even lead to teratogenic effects as shown with fish embryos; all physiological responses to HS-mediated stress require energy, which were compensated on the expense of yolk as shown with zebra fish embryos. One Finnish field survey supports the view of a strong chemical stress, as the weight yield in fish species decreases with increasing HS content in the lakes. DISCUSSION: HS exert a variety of stress symptoms in aquatic and compost organisms. According to current paradigms of ecotoxicology, these symptoms have to be considered adverse, because their compensation consumes energy which is deducted from the main metabolism. However, the nematode C. elegans looks actively for such stressful environments, and this behavior is only understandable in the light of new paradigms of aging mechanisms, particularly the Green Theory of Aging. In this respect, we discuss the mild HS-mediated stress to aquatic and compost organisms. New empirical findings with HS themselves and HS building blocks appear to be consistent with this emerging paradigm and show that the individual lifespan may be expanded. At present the ecological consequences of these findings remain obscure. However, a multiple stress resistance may be acquired which improves the individual fitness in a fluctuating environment. CONCLUSIONS: It appears that dissolved HS have to be considered abiotic ecological driving forces, somewhat less obvious than temperature, nutrients, or light. PERSPECTIVES: The understanding of the ecological control by dissolved humic substances is still fragmentary and needs to be studied in more details. PMID- 18380233 TI - Effect of herbicides on glutathione S-transferases in the earthworm, Eisenia fetida. AB - AIM AND BACKGROUND: Earthworms have been studied as a readily available, easily maintainable and cheap test species for assessing chemical pollution, and may be an alternative to in vivo rodent bioassays. The current investigation aims to characterize detoxification enzymes in Eisenia fetida and stress response against two herbicides with different modes of action, namely, fenoxaprop and metolachlor. METHODS: Herbicides were applied to soil containing earthworms. Animals were then collected, sacrificed and shock-frozen. Extracted protein was analyzed for glutathione S-transferase (GST) activity using CDNB (1-chloro-2,4 dinitrobenzene), DCNB (1,2-dichloro-4-nitrobenzene), pNBC (p nitrobenzylchloride), PNOBC (p-nitrobenz-o-ylchloride) and selected herbicides. GST isoenzymes were partially purified by affinity chromatography and molecular weights were estimated by SDS-PAGE. RESULTS: In E. fetida protein extracts, GST activity towards model compounds ranked as CDNB>DCNB>PNBOC>PNBC. Fluorodifen was not conjugated at all, but fenoxaprop and metolachlor were conjugated at low rates. Furthermore, the GST isoenzyme pattern changed during the incubation with herbicides, either due to stress or as a defense reaction. After incubation with monochlorobimane, a strong fluorescence of the intestinal tract and the intersegments was observed, indicating organ-specific GST induction. DISCUSSION: According to the author's knowledge, here, for the first time, evidence is presented that E. fetida GST are also capable of conjugating a wider range of xenobiotic substrates. Different forms of GST were observed and changes in GST isoforms due to the herbicide treatment were also noticed. GST conjugation rates varied between different herbicides used in this experiment. It might be assumed that herbicides may well be detoxified by earthworms, to a certain extent, but that they are also potent stress factors influencing the detoxification system of the animal. High doses or long exposure might lead to deleterious effects on earthworms and limit their survival rate. The use of the animals as bioindicators for herbicides and herbicide residues seems very promising, but is surely influenced by the lack of detoxification for some compounds. CONCLUSIONS: Conjugation of several xenobiotics with model substances and herbicides is proven in the earthworm E. fetida. However, E. fetida has only limited capabilities of detoxifying herbicidal compounds. Different isoforms of GST were involved and altered in their activity after treatment. RECOMMENDATIONS AND PERSPECTIVES: The accumulation of GS-conjugates and their determination via fluorescence microscopy is a quick and secure, additional marker for exposure that should be further developed to complement existing biotests. The described methods and endpoints might help to understand the complex reaction of earthworms towards herbicides and lead to an adapted test methodology. PMID- 18380234 TI - Respiratory health of road-side vendors in a large industrialized city. AB - GOAL, SCOPE AND BACKGROUND: The literature reports a high prevalence of respiratory symptoms associated with exposure to motor vehicle exhaust emissions and people exposed to vehicle emissions are at risk of reduced lung function and cardiovascular performance. Although the effect of traffic emissions is a known risk to respiratory health, retailers are often situated along major roads in a busy urban environment to maximise customers. Shop assistants in an air conditioned environment should be less exposed to traffic fumes and their lung function should be better preserved compared to 'unprotected' vendors exposed directly to vehicle emissions. The lung function of these two groups of workers has not previously been compared. The aims of this study are to determine if there is a difference in the concentration of respirable particles of diameter less than 10 micrometers (PM10) inside and outside air-conditioned shops along a busy major road; and to compare the lung function parameters, blood lead levels and respiratory symptoms between road-side vendors directly 'exposed' to traffic fumes and vendors working in 'protected' air-conditioned shops along the same road. METHODS: Roadside vendors (n=33) and adjacent shop assistants (n=31) were recruited for the study in a district known to have high emissions and pollution measures. All subjects were asked to complete a questionnaire providing the frequency of their respiratory symptoms. Spirometry pulmonary function tests were conducted and exhaled carbon monoxide levels (ECO), oxygen saturation and blood lead levels (BLL) were measured. PM10 level was concurrently measured in the subject's working environment. Pulmonary function data and ECO levels collected from a cohort of university staff aged > or = 30 years (n=92) at a university campus were included for comparison. RESULTS: The concentration of particulate matter (PM10) at the roadside (210 +/- 70 microg/m3) was significantly higher than inside the shops (130 +/- 40 microg/m3). There was no difference in lung function parameters or BLL between the groups, however the forced vital capacity of both groups of vendors was significantly lower than a 'control' cohort of academic institution personnel (n=92). DISCUSSION: This study illustrated that while the level of pollution improved with air conditioning, this may not be sufficient to prevent respiratory consequences. Our data suggests that a reduction in occupational exposure is not enough to protect these workers. CONCLUSIONS: Air-conditioned work environment is insufficient to protect the respiratory system against the adverse effects of exposure to vehicle emissions. RECOMMENDATIONS: The overwhelming effect of exposure during travel to and from work and at home cannot be underestimated. More stringent guidelines to control pollution appear necessary to protect the respiratory health of both shop-keepers and road-side vendors and further studies to explore the effect on respiratory health of the exposure to pollutants associated with commuting to and from work are warranted. PMID- 18380235 TI - U and Th in some brown coals of Serbia and Montenegro and their environmental impact. AB - GOAL, SCOPE AND BACKGROUND: The objective of this paper is to determine and compare the concentrations of U and Th in soft to hard brown (lignite to sub bituminous) coals of Serbia and Montenegro. It also presents comparison of the obtained data on U and Th concentrations with the published data on coals located in some other countries of the world. Almost the whole coal production of Serbia and Montenegro is used as feed coals for combustion in thermal power plants. METHODS: Channel samples from open pit and underground mines and core samples were collected for hard and soft brown coals. For the analysis the samples were decomposed using microwave technique. Obtained solutions containing U and Th were analyzed by inductively coupled plasma mass spectroscopy (ICP-MS) using NIST standards. RESULTS: Concentration of U from the investigated basins and the corresponding mine fields ranges within 0.60-70.10 mg/kg, 0.65-3.20 mg/kg, 0.95 6.59 mg/kg, 1.20-6.05 mg/kg, 0.80-6.66 mg/kg, 0.18-89.90 mg/kg, 0.19-4.14 mg/kg, and 0.28-3.52 mg/kg for the Kostolac, Kolubara, Krepoljin, Sjenica, Soko Banja, Bogovina East field, Senje-Resavica and Pljevlja basins, respectively. Concentration of Th ranges within 0.20-2.60 mg/kg, 0.84-6.57 mg/kg, 1.48-6.48 mg/kg, 0.12-2.71 mg/kg, 0.13-4.95 mg/kg, 0.14-3.48 mg/kg, 0.29-3.56 mg/kg, and 0.17-1.89 mg/kg for the Kostolac, Kolubara, Krepoljin, Sjenica, Soko Banja, Bogovina East field, Senje-Resavica and Pljevlja basins, respectively. DISCUSSION: Brown coal from Senje-Resavica, Kolubara, Kostolac and Pljevlja is characterized by low U concentration. Coals form the Krepoljin, Soko Banja and Sjenica basins have slightly higher U concentrations than the mentioned group. The highest concentration of U is characteristic for the coal from the Bogovina East field. Concentration of Th in coals from Serbia and Montenegro has proved to be low. Out of all investigated coal basins, only the coal from the Krepoljin and Kolubara basins has high concentration of Th. The hydrothermally altered rocks of the Timok dacite-andesite complex, representing the basement of the Bogovina basin, could be a potential source of U, especially at the bottom part of the Lower seam of the Bogovina East field. CONCLUSIONS: This study shows that brown coals in Serbia and Montenegro (soft to hard brown coals or lignite to sub bituminous) contain low levels of U (5.30 mg/kg, average value and 2.10 mg/kg geometric mean value) and Th (1.80 mg/kg, average value and 1.12 mg/kg geometric mean value). There are some obvious differences in concentration of U and Th in coals from different basins in Serbia and Montenegro. The approximate value for U and Th release mainly from power plants was 644.33 t and 983.46 t, respectively within the period 1965-2000 for the studied mines in Serbia, and 23.76 t and 15.05 t for the Potrlica mine (Montenegro) within the period 1965-1997. RECOMMENDATIONS: The coals in Serbia and Montenegro show no identifiable unfavourable impact on the surrounding environment, due to low natural radioactive concentration of U and Th, but further investigations concerning human health should be performed. PERSPECTIVES: Preliminary research revealed that in some Serbian coals (and, particularly, parts of the coal seam) U and Th content are rather high. Such coals should be carefully studied, as well as U and Th concentrations in ash, fly ash, waste disposals, nearby soil and ground water. Further studies should include determination of the radioactivity of all these products, and estimation of possible health impact. PMID- 18380236 TI - Impact of a smelter closedown on metal contents of wheat cultivated in the neighbourhood. AB - BACKGROUND: The contamination of soils by heavy metals engenders important environmental and sanitary problems in Northern France where a smelter has been located for more than one hundred of years. It has been one of the most important Pb production sites in Europe until its closedown in March 2003. Ore smelting process generated considerable atmospheric emissions of dust. Despite an active environmental strategy, these emissions were still significant in 2002 with up to 17 tonnes of Pb, 32 tonnes of Zn and 1 tonne of Cd. Over the years, the generated deposits have led to an important contamination of the surrounding soils. Previous studies have shown pollutant transfers to plants, which can induce a risk for human and animal health. The objective of this study was to evaluate the consequences of the smelter closedown on the Cd and Pb contents of wheat (grain and straw) cultivated in the area. METHODS: Paired topsoil and vegetable samples were taken at harvest time at various distances to the smelter. The sample sites were chosen in order to represent a large range of soil metal contamination. Sampling was realised on several wheat harvests between 1997 and 2003. 25 samples were collected before the smelter closedown and 15 after. All ears of about 1 m long of two rows were manually picked and threshed in the lab. Similarly, straw was harvested at the same time. Total metal contents in soil and wheat samples were quantified. RESULTS: A negative correlation between metal concentrations in soil and the distance to the smelter was shown. The wheat grain and straw showed significant Cd and Pb contents. The straw had higher metal contents than the grain. During the smelter activity, the grain contents were up to 0.8 mg kg(-1) DM of Cd and 8 mg kg(-1) DM of Pb. For the straw, maximum contents were 5 mg kg( 1) DM of Cd and 114 mg kg(-1) DM of Pb. After the smelter closedown, we observed a very large decrease of Pb in the grain (82%) and in the straw (91%). A smaller decrease was observed for Cd in grain. Despite this improvement, 80% of the studied samples remained non-acceptable for human consumption, according to the European legislation values, due to a high Cd content. DISCUSSION: Results highlighted a difference in metal accumulation in the plant organs as well as a difference in metal uptake. The approach pointed out the importance of atmospheric fallout in the wheat contamination pathways for Pb. The smelter closedown has lead to a decrease of the Pb content in wheat. It is interesting to relate this finding with the lead blood levels in children living close to the smelter. CONCLUSIONS: Those results have confirmed the importance of dust fallout in the plant contamination pathways. Before the closedown, Pb measured in the plant was principally originating from the smelter dust emissions. It raised the question of the sanitary risks for humans and animals living in the surrounding a of the smelter. RECOMMENDATIONS AND PERSPECTIVES: In the literature, very few articles take the dust deposit as contamination pathways for crops into consideration. However, in highly contaminated sites, this pathway can be very important. Thus, it would be worthy studying the uptake of metal contaminants by plants through the foliar system. PMID- 18380237 TI - Business continuity planning. It's a critical element of disaster preparedness. Can you afford to keep it off your radar? PMID- 18380238 TI - Forged in fire. A San Diego hospital benefits from automated inventory management during the 2007 wildfires. PMID- 18380239 TI - Automating anatomic pathology. Implementing an AP solution that integrates with your hospital's LIS can improve workflow and productivity. PMID- 18380240 TI - The evidence-based advantage. The proper use of EBM in CPOE can enhance physician decision-making and improve patient outcomes. PMID- 18380241 TI - The critical link. Remote monitoring of ICU patients can save lives and decrease patient length of stay across rural America. PMID- 18380242 TI - Jumping to solutions. Implementing a structured "look before you leap" discovery process can lead to successful IT adoption. PMID- 18380243 TI - Achieving effective biosurveillance. PMID- 18380244 TI - The sins of specialists. AB - In medieval times, moralists compiled lists of sins that certain types of persons, such as clerics, lawyers, kings and physicians, were prone to commit. These persons were set aside from the ordinary crowd by their special duties, skills and knowledge. Today, we might call them "specialists" or "professionals". These sins reflected failures to attain the ideals that the specialists were held to. Among the most common sins were actions that took advantage of those who depended upon the specialist's ability to respond to calls for help. While the lists of sins did not include dentists, who did not exist as specialists at that time, this lecture will pursue the theme of the medieval moralists into the time when dentistry emerges as a specialty form of care. In essence, it asks how professional ethics protects patients or clients from exploitation. PMID- 18380245 TI - Professional ethics and professional etiquette in dentistry: are they compatible? AB - In keeping with the theme of this colloquium, two aspects of ethics in dentistry are addressed: its evolution and its future. With respect to its evolution, two examples of changes in the design of clinical trials in dentistry are discussed. These concern the current requirement of informed consent from the subjects in the trial, now taken for granted, but not necessarily observed before 1964. The Vipeholm dental caries study is one example of pre-Helsinki Declaration experimentation. The second example, also drawn from caries research design, concerns the stricture on the use of placebo-controlled trials in the face of a proven drug. For example, the design of clinical trials of fluoride dentifrices has evolved since the mid 1970s. The use of a placebo-inactive control group is no longer acceptable as it would deprive its subjects of a proven caries preventive agent and would expose its subjects to increased caries risk. While definitions of professional ethics in dentistry may vary, the ADA Code of Ethics includes five principles: patient autonomy ("self-governance"), non-maleficence ("do no harm"), beneficence ("do good"), justice ("fairness") and veracity ("truthfulness"). Professional etiquette refers to the way dentists relate to one another and is governed by the ADA Code of Professional Conduct which expresses specific types of conduct that are either required or prohibited. Sometimes, ethics and etiquette may conflict. The problem of financial issues that conflict with ethical ones is discussed along with the problem of commercialism in the practice of dentistry. Debts from dental school may adversely affect the professional behavior of young dentists, while general dentists might succumb to "goodies" provided by specialists. These often include continuing education courses, gifts, trips, and kickbacks. Specialists may fail to inform patients of improper or poor quality treatment by the referring general practitioner, fearing loss of referrals. Of course these issues are not unique to dentistry; they apply to all health care providers. Nor is this a new problem, but it requires fixing for the future of ethical dental practice. PMID- 18380246 TI - What have teeth taught us about culture? Practice, patienthood and ethics in the history of dentistry and public health. AB - Teeth cut across cultures. They transgress cultural boundaries but also define social boundaries. They provide information about what goes into the mouth, and what the mouth is appropriately used for. Scrutiny of teeth identified a new biomedical space to analyze pain, and created a new culture of medicine for such practices. This paper uses the evolution of dentistry since the 18th century to look at how our social and scientific understanding of teeth has shaped cultural attitudes about pain, politics, beauty and prophylaxis. It then raises questions about how these attitudes in turn create ethical contexts for the practice of dentistry around the world. PMID- 18380247 TI - Ethics versus legal informed consent--a distinction with little difference. AB - The core principles of dental ethics and legal standards of care have similar foundations. Both are dedicated to place the patient's best interest as primary and the practitioner's interest as secondary, Similarities between ethics and the law demonstrate that most often there may be distinctions but little core differences. Informed consent principles illustrate the comparison between dental ethics and the law. PMID- 18380248 TI - Early struggles to identify ethical standards in dentistry: Dr. Benjamin Brown and the amalgam war of the 1840s. AB - Dr. Benjamin Boyer Brown was one of the leading physicians and dentists in St. Louis during the 1830s and 1840s as well as one of its most esteemed citizens for his charitable and educational works. He was also one of the founders of organized dentistry, first editor of the Dental Register of the West, as well as a respected researcher and educator in dentistry, and a member of the American Society of Dental Surgery, a forerunner of the American Dental Association. This society, declared the use of amalgam to be not only unethical but malpractice, and members were forced to sign a pledge not to use it. Although many dentists opposed this decision and ignored the pledge altogether, Dr. Brown was morally unwilling to remain quiet. He vocally opposed the decision of his colleagues to ban amalgam on ethical grounds. In spite of his appeal for reason and his high profile, he was one of the few dentists to be expelled from organized dentistry. He moved to California during the height of the gold rush to begin a new life. Dr. Brown's experience illustrates several issues in dental ethics that remain with us today. PMID- 18380249 TI - Painless Parker's legacy: ethics, commerce, and advertising in the professions. AB - This presentation will review the life and contributions of Dr. Edgar Parker, the infamous and controversial pioneer who specialized in a precarious straddling of the ethics of the commercial marketplace and the ethics of care. Something of a Rorschach test, he was alternatively referred to as a charlatan, the first people's dentist, a renegade, a crusader, a quack, the Henry Ford of dentistry, and "a menace to the dignity of the profession". He eventually owned and managed thirty dental offices, several in San Francisco, as well as the Parker Dental Circus. Because many young, twenty-first century practitioners have little problem with slick advertising, it seems appropriate to revisit Painless Parker's career and contribution to the current state of affairs. PMID- 18380250 TI - An ethical lesson learned from the equestrian sculpture, "The Torch Bearers," at the University of Madrid Dental School. AB - As dental professionals, we continue to learn ethical lessons throughout our careers and beyond. We may experience them in our day-to-day involvements with peers and patients, or they may present themselves under more unusual circumstances. In October, 1974, at the University of Madrid Dental School, the senior author had a deeply impressionable experience minutes before he helped to present a course to Spanish dentists in operative dentistry. His co-presenter was Dr. Miles R. Markley of Denver, Colorado, a National Consultant in Restorative Dentistry for the U.S. Air Force. Their ethics teacher that day was Dr. Gerardo Zabalo, Dean of the dental school. His object lesson was delivered at the base of an equestrian, aluminum, larger-than-life sculpture, "The Torch Bearers" which faced the school. The sculptor shows an old, wrinkled man, lying prostrate, with only enough strength left to raise a lighted torch upward. Reaching down to receive the torch is a robust, energized young man, who is sitting upon a sturdy stallion. The lesson which Dean Zabalo taught that day was simple but profound. The lit torch of ethics, learning and values is essentially transmitted from generation to generation, as young students learn from their elders, accept the torch from their retiring leaders, keep the torch brightly lit as they mature in their profession, and, then in turn, hand it over to those young professionals who will replace them. This process will continue, age after age. Thus the torch of learning and thinking is passed on. In the world of education, human contact and the continued sanctity of the scientific written word must be paramount. Students must be taught how to read, learn and think. Computer technology used exclusively as a mode of teaching and learning can never become a viable substitute for our traditional interactive methods which emphasize human to human interaction. PMID- 18380251 TI - Dental ethics in a larger context: one point of view. AB - Since the end of World War II, the practice of dentistry has been largely transformed from a "calling" into a cog in the ever-expanding "Healthcare Industry". In the process, the distinction between professional ethics and the ethics of commerce has been attenuated and, to a large extent, lost. Today's dentist is faced with an inherent conflict between the pledge of the health professional to hold the patient's interests primary (and above all, to do no harm), and the self-protective commercial principle of caveat emptor. Pressures towards commercialism come from the government and the insurance industry, the increasingly unfavorable ratio between professional fees and the cost of production, and the high cost of dental education. Viewed simplistically, much of dentistry today has an outward form resembling commodity production. Recognizing the substantial forces tending to attenuate ethical standards in our profession may aid us in resisting their encroachments. PMID- 18380253 TI - Futurists ponder the possibilities. PMID- 18380252 TI - Readers' perspectives. Payer organizations accept and appropriately process HIPAA transactions. PMID- 18380254 TI - So you want to be a CIO? PMID- 18380255 TI - Finding the right I.T. PMID- 18380256 TI - Taking on a new task. PMID- 18380257 TI - Starting from scratch. PMID- 18380258 TI - Sharing the costs of e-records. PMID- 18380259 TI - Plan fine-tunes subsidy program. PMID- 18380260 TI - Why use electronic records? PMID- 18380261 TI - Evaluating teaching strategies: a blended perspective. PMID- 18380262 TI - Nursing student caring behaviors during blood pressure measurement. AB - The purpose of this multisite, nonexperimental study was to examine, using a repeated measures design, the effects of a teaching intervention designed to promote caring behaviors as students learn the psychomotor skill of blood pressure measurement. Watson's theory of human caring and a combination of cognitive and connectionist learning theories were used as the organizing construct. Baccalaureate nursing student participants were videotaped and evaluated at two points in time while performing the psychomotor skill of blood pressure measurement on a role-player. Role-players rated the students' caring behaviors using the Role Player Survey of Caring Behaviors During Blood Pressure Measurement instrument. Between these data collection points, students learned about caring behaviors through analysis of a videotaped role-play and required readings. An evaluator randomly selected 10 student videotapes from each of the 6 baccalaureate nursing program study sites and noted the presence or absence of caring behaviors on the Caring Behaviors During Blood Pressure Measurement instrument. Pretest and posttest scores on both subjective and objective research instruments were compared using descriptive statistics and repeated measures analysis of variance (ANOVA). Students demonstrated a significant improvement in objective and subjective caring behaviors between the two performance examinations. The findings support further investigation of teaching interventions to promote the development of caring behaviors during nursing psychomotor skill development. PMID- 18380263 TI - The Omaha System and quality measurement in academic nurse-managed centers: ten steps for implementation. AB - Measuring and describing client problems, nursing interventions, and outcomes is a challenge in nursing care. This study reviews the literature about the use of the Omaha System and describes the steps used to introduce and implement the Omaha System as a documentation and outcome measurement system in academic nurse managed centers. The goal of the article is to provide a guide for nurse educators to develop and implement a quality measurement system that can be used in clinical settings by undergraduate students. The Omaha System is a strategy to introduce and incorporate evidence-based practice in the undergraduate nursing clinical experience. PMID- 18380264 TI - Team-based learning in an undergraduate nursing course. AB - To increase student participation in the learning process, active learning methods, including small group learning, have become increasingly popular in modern curricula. One kind of small group learning, team-based learning, is a relatively new instructional strategy in health care education. Team-based learning uses theoretically based and empirically grounded strategies for ensuring the effectiveness of small groups working independently in classes with high student-to-faculty ratios (e.g., up to 200:1), without losing the benefits of faculty-led small groups with lower ratios (e.g., 7:1). To explore the effectiveness of this learning pedagogy, we evaluated students' level of engagement and attitudes toward the value of teams. Findings demonstrated that team-based learning is an effective teaching strategy for large groups of students. PMID- 18380265 TI - Students' opinions about and expectations of effective nursing clinical mentors. AB - Nursing practice is an integral part of nursing education in which clinical mentors play an important role. Mentors' attitudes, experience, and knowledge influence nursing students and help them to complete their nursing practice with success. The purpose of this qualitative study was to determine nursing students' opinions about and expectations of effective nursing clinical mentors. Focus group interviews were used to obtain data from 24 students after completion of their third year of nursing school. The students, who had nursing practice experience in internal diseases, surgery, gynecology, pediatrics, psychiatry, and public health, were divided into three groups of 8 students. The students noted that effective clinical mentors should be able to communicate without prejudice, give positive feedback, have empathy, require students to do their own research, and offer the students information. PMID- 18380266 TI - Dimensions of caring: a qualitative analysis of nurses' stories. AB - The purpose of this qualitative, phenomenological study is to describe dimensions of caring as they relate to and clarify the practice of professional nursing. Nurses are unique caregivers, and their work at the bedside and in the community matters. What nurses do as they care for patients is multi-dimensional, complex, and essential. Two hundred stories written by nurses were analyzed using Giorgi's methodology for existential phenomenology. Their stories indicate that nursing goes far beyond technical skills. Seven dimensions of caring that define professional nursing practice were found: caring, compassion, spirituality, community outreach, providing comfort, crisis intervention, and going the extra distance. The nurses' stories demonstrate that the dimensions of caring that define professional nursing practice are universal. Documentation of nurse's stories facilitates reflective and thoughtful practice, while clarifying the essential components of nursing. PMID- 18380267 TI - Creating a home care simulation laboratory. AB - Simulation laboratories are being created to prepare students and new staff for clinical experiences. With puppets and garage sale objects, a life-in-home laboratory was developed to meet the needs of those first encountering a home care situation. PMID- 18380268 TI - Integrating the curriculum to advance baccalaureate nursing education. AB - In 2004, the School of Nursing at the University at Buffalo began a program in which individuals with a degree in another field could complete an intensive 12 month program leading to a baccalaureate degree in nursing. Curriculum design using courses from the basic baccalaureate and RN-to-BS program, as well as graduate courses, not only provided the opportunity to integrate accelerated bachelor of science (ABS) students with the other student populations but eliminated the need for the development of courses specific to the ABS program. A unique feature of the program is the incorporation of graduate courses, allowing students to earn 9 to 12 graduate credits. The potential for future nursing leaders has also increased, given the characteristics of these mature, highly motivated, career-minded students who have chosen nursing as a second career. PMID- 18380269 TI - Comparing the academic performance of students in traditional and second-degree baccalaureate programs. AB - Programs offering accelerated baccalaureate (BSN) curricula to students with bachelor's degrees in other fields are growing in popularity. Such students' academic ability may differ from that of students pursuing the BSN as their first degree, due to academic maturity, greater confidence, and polished study skills. This study directly compared accelerated second-degree BSN and traditional BSN students under controlled conditions matched for identical instruction and performance measures. Outcomes analyzed included class test scores, nationally standardized examination scores, skills laboratory performance, and final course grades. Age, as a possible factor of any such differences, was also analyzed. The accelerated students performed significantly higher on every measure than did the students in the traditional program. Age was not found to be predictive of success in either group. Recommendations include modification of teaching strategies to accommodate accelerated students capable of higher performance. PMID- 18380270 TI - Using a social constructivist model of teaching to create a learning community. PMID- 18380271 TI - Migraine and chronic daily headache--we could be doing better. PMID- 18380272 TI - Donation after cardiac death: how best to address ethical concerns. PMID- 18380273 TI - A neuromuscular approach to statin-related myotoxicity. AB - Approximately 95% of statin-treated patients tolerate this form of cholesterol management without any adverse effects. However, given their efficacy in reducing low density lipoproteins and cardiovascular events large numbers of patients are selected for statin therapy. Therefore muscle complications are, in fact, quite common. Limited understanding of the underlying pathophysiology has hampered physicians' ability to identify patients at risk for developing statin myotoxicity. A growing number of published case reports/series have implicated statins in the exacerbation of both acquired and genetic myopathies. A clinical management algorithm is presented which outlines a variety of co-morbidities which can potentiate the adverse effects of statins on muscle. In addition, a rational approach to the selection of those patients most likely to benefit from skeletal muscle biopsy is discussed. Ongoing work will define the extent to which statin-intolerant patients represent carriers of recessive metabolic myopathies or pre-symptomatic acquired myopathies. The expanding importance of pharmacogenomics will undoubtedly be realized in the field of statin myopathy research within the next few years. Such critical information is needed to establish more definitive management and diagnostic strategies. PMID- 18380274 TI - Seizure semiology: value in identifying seizure origin. AB - The diagnosis of epilepsy depends upon a number of factors, particularly detailed and accurate seizure history, or semiology. Other diagnostic data, consisting of electroencephalography, video-monitoring of the seizures, and magnetic resonance imaging, are important in any comprehensive epilepsy program, particularly with respect to lateralizing and localizing the seizure focus, if such a focus exists, and with respect to determining the type of seizure or seizure syndrome. The aim of this review is to present a survey of important semiologic characteristics of various seizures that provide the historian with observations, which help to lateralize and localize epileptic zones. Clinical semiology is the starting point of understanding a seizure disorder and making the diagnosis of epilepsy. While it may not provide unequivocal evidence of localization of the epileptic focus, nevertheless it usually directs subsequent investigations, whose concordance is necessary for the ultimate localization. PMID- 18380275 TI - Skeletal muscle metabolic dysfunction in obesity and metabolic syndrome. AB - Obesity and the related metabolic syndrome have become a worldwide epidemic. Inactivity appears to be a primary causative factor in the pathogenesis of this obesity and metabolic syndrome. There are two possible, perhaps not mutually exclusive, events that may lead to intramyocellular lipid accumulation and mitochondrial dysfunction in patients with obesity. First, obesity, with high intake-associated lipid accumulation in muscle may interfere with cellular mitochondrial function through generation of reactive oxygen species leading to lipid membrane peroxidative injury and disruption of mitochondrial membrane dependent enzymes. This in turn leads to impaired oxidative metabolism. Secondly, a primary defect in mitochondrial oxidative metabolism may be responsible for a reduction in fatty acid oxidation leading to intramyocellular lipid accumulation as a secondary event. Non-invasive techniques such as proton (1H) and phosphorus (31P) magnetic resonance spectroscopy, coupled with specific magnetic resonance imaging techniques, may facilitate the investigation of the effects of various ergometric interventions on the pathophysiology of obesity and the metabolic syndrome. Exercise has positive effects on glucose metabolism, aerobic metabolism, mitochondrial density, and respiratory chain proteins in patients with metabolic syndrome, and we propose that this may be due to the exercise effects on AMP kinase, and a prospective physiological mechanism for this benefit is presented. A physiological model of the effect of intramyocellular lipid accumulation on oxidative metabolism and insulin mediated glucose uptake is proposed. PMID- 18380276 TI - Methylprednisolone for acute spinal cord injury: 5-year practice reversal. AB - OBJECTIVE: To re-evaluate practice patterns for methylprednisolone (MP) administration in patients with acute spinal cord injury (SCI) within the spinal surgery community across Canada five years after the publication of practice recommendations. METHODS: Canadian orthopedic and neurological spine surgeons were surveyed at their respective annual meetings about their practice of steroid administration for acute SCI by means of a questionnaire comprised of the same seven questions posed five years ago plus an additional question related to change of view. RESULTS: Forty-two surgeons and twenty-one residents directly involved in the acute management of SCI completed the questionnaire. Seventy-six percent of spinal surgeons do not prescribe MP for SCI in sharp contrast to 76% who prescribed it five years ago. Of the 24% who use steroids, the NASCIS II dosing regimen is most commonly followed. One third of physicians continue to administer MP because of fear of litigation. CONCLUSIONS: Over a five year period there has been a complete reversal in practice patterns of MP administration for SCI, along with an increased familiarity of the published literature. Attendance at meetings, participation in local group discussions, and peer-reviewed publications appear effective in altering practice preferences arising from peer pressure and even fear of litigation. PMID- 18380277 TI - Multidisciplinary treatment for headache in the Canadian healthcare setting. AB - BACKGROUND/OBJECTIVES: Multidisciplinary treatment programs are seen as an effective way to treat patients with chronic illness. The purpose of this study was to describe a multidisciplinary headache program which was developed in the Canadian public healthcare setting, and to report on patient perceptions of the program and patient treatment outcomes. METHODS: The Calgary Headache Assessment and Management Program (CHAMP) was developed with initial funding from Alberta Health, and continued with function from the Calgary Health Region. Patient perceptions of the program were obtained with questionnaires. Outcome measures for a cohort of patients who completed the Self-Management Workshop were obtained using standard headache related disability measures. RESULTS: Patient perceptions of the education session, the Lifestyle Assessment, and the Self-Management Workshop were very positive. Headache Disability Inventory scores fell from 56.2 to 46.3 from baseline to three months post Self-Management Workshop (p < .001). Corresponding scores for the HIT-6 were 63.6 and 58.2 (p < .001). CONCLUSIONS: Multidisciplinary headache treatment programs can be developed in the Canadian public healthcare system. The program described here was well accepted by many patients and perceived to be useful by them. Headache related disability as measured by standard measures was significantly reduced after participation in the program. PMID- 18380278 TI - 3D conformal radiotherapy and cisplatin for recurrent malignant glioma. AB - PURPOSE: To determine the maximum tolerated dose of 3D conformal radiotherapy in combination with Cisplatin for patients with recurrent malignant gliomas. METHODS: From 1999-2003, nine patients with recurrent malignant glioma received fractionated radiotherapy and Cisplatin (20 mg/m2/d IV on days 1-5) in a Phase I radiation dose escalation trial. Three sequential dose levels were evaluated: 25 Gy, 30 Gy, and 35 Gy, using 5 Gy fractions. All patients received prior external beam radiation (median dose 59.4 (20-60) Gy) and five patients received prior chemotherapy. RESULTS: Six male and three female patients were enrolled with a median age of 52 years, and a median Karnofsky performance status score of 70. The median re-irradiated tumor volume was 18.9 (0.1-78.5) cm3 and the median follow-up was 8.8 (3.2-31.2) months. One patient (30 Gy/ 6 fractions) experienced medically reversible acute grade 3 toxicity. A second patient (35 Gy/ 7 fractions) experienced acute grade 2 toxicity and histology showed tumor and radiation effect. A third patient (25 Gy/ 5 fractions) experienced late grade 3 toxicity from radiation necrosis. The radiological responses consisted of complete response (1 patient), partial response (1 patient), and stable disease (2 patients). The median overall survival was 8.8 months (95% CI 8.0-9.9), and the median disease free interval was 2.0 months (95% CI 1.4-4.4). Seven patients received chemotherapy following re-irradiation and Cisplatin. CONCLUSION: The maximum tolerated dose of 3D conformal fractionated radiotherapy was 30 Gy in 6 fractions with low dose Cisplatin, which was well tolerated in terms of acute toxicity for our patient population. This regimen demonstrated only modest efficacy in the treatment of recurrent malignant glioma. Combinations of conformal re-irradiation and other systemic agents may merit investigation. Currently our recommended dose is 30 Gy in 6 fractions for selected patients. PMID- 18380279 TI - Postural sway and effect of levodopa in early Parkinson's disease. AB - OBJECTIVE: To characterize postural stability control and levodopa responsiveness in early Parkinson's disease (PD). METHODS: Postural sway was studied during quiet stance in ten patients within six years of PD onset, both before (OFF) and after (ON) regular oral levodopa dosing. Postural sway was recorded using a force platform during 30 sec with eyes open, and six dependent variables were examined. RESULTS: Mild baseline subclinical changes in postural sway were recorded in our patients. Clear benefit was observed in five out of six characteristics (mean sway, transversal sway, sagittal sway, sway intensity, and sway area) in the ON condition. CONCLUSION: Postural control mechanisms are affected early in PD and modulated by dopamine. PMID- 18380280 TI - The relation of ultraviolet radiation and multiple sclerosis in Newfoundland. AB - BACKGROUND: It has been thought that the occurrence of multiple sclerosis (MS) could be associated with daily ultraviolet exposure. In this study we investigated the geospatial association between average daily ultraviolet B (UVB) irradiance and MS prevalence in Newfoundland and Labrador (NL), Canada. METHODS: A complete list of patients diagnosed with MS in the province of NL was constructed. Places of habitation from birth to diagnosis were ascertained by mailout survey. RESULTS: A 74% rate of return on the survey results was obtained. A plot of the average daily erythemal UV over the available five years (1998 2002) shows that the distribution of MS follow a north-south gradient. Average daily UVB measurements are lower in the higher latitudes. A statistically significant negative correlation of MS incidence with erythemal UVB was found that is stronger than the correlation using latitude. This correlation appears to be strongest in the first year of life and declines when subsequent years are examined up to age ten. No significant correlation was found for the subjects' locale of habitation at the time of their first MS attack. CONCLUSIONS: This study suggests that UVB radiation may contribute to the pathogenesis of MS. PMID- 18380281 TI - The learning curve of minimally-invasive lumbar microdiscectomy. AB - OBJECT: The safe integration into practice of a new surgical technique requires an appreciation of the learning curve. The object of this study was to assess the learning curve for minimally invasive microdiscectomy (MIM) utilizing a tubular retractor system. METHODS: A prospective evaluation of a single surgeon's first 52 consecutive MIM cases for radiculopathy secondary to single-level posterolateral lumbar disc herniation was performed. The learning curve was assessed using operative time, conversion to open rate, complications, and length of hospitalization. RESULTS: The duration of operative time decreased over the course of the study (range, 49-151 min). By case 15, operative time was typically 60 min or less. There was only one conversion to an open procedure (Case 2). Complications occurred in three cases. All but nine patients were discharged home on the day of surgery. CONCLUSION: The learning curve for MIM was demonstrated. Further assessment of this curve for a large group of surgeons is necessary before a randomized controlled trial comparing standard microdiscectomy to MIM can be conducted. PMID- 18380282 TI - Complications in 622 cases of frame-based stereotactic biopsy, a decreasing procedure. AB - BACKGROUND: Frame-based stereotactic brain biopsy has played an important role in the management of patients with suspected neoplastic intracranial lesions over the last three decades. We reviewed the surgical experience of one surgeon to determine the nature and frequency of complications associated with this procedure. METHODS: Records were reviewed for 858 patients undergoing frame-based stereotactic procedures from January 1986 to May 2006. Data on each case were prospectively collected by the senior author. Procedures for Ommaya reservoir placement, brachytherapy, stereotactic craniotomy flap localization, shunt placement, or treatment of previously-diagnosed intracranial cystic lesions were excluded, leaving 614 patients in whom a total of 622 procedures were performed for purely diagnostic purposes. Complication rates and their association with clinical variables were sought. RESULTS: Morbidity and mortality rates were 6.9% (43/622) and 1.3% (8/622), respectively. The risk of symptomatic hemorrhage (intracerebral hemorrhage [ICH], subarachnoid hemorrhage [SAH], intraventricular hemorrhage [IVH]) was 4.8%. The risks of transient or permanent neurological deficits were 2.9% (18/622) and 1.5% (9/622), respectively. Biopsy of deep-seated lesions was associated with increased overall complication rate, while biopsy of Glioblastoma Multiforme (GBM) was associated with perioperative mortality. CONCLUSIONS: Overall, complication rates were comparable with those in previous reports. The subgroup of patients with deep-seated lesions or a histologic diagnosis of GBM may possess an elevated risk of overall complications or mortality, respectively, compared to other patients undergoing frame-based stereotactic brain biopsy. PMID- 18380283 TI - ONF trainee awards contribute to capacity building in neurotrauma. AB - OBJECTIVE: Injury to the brain and spinal cord is one of the most catastrophic and costly occurrences in the Ontario health system. The objective of the present study was to evaluate the impact of past Ontario Neurotrauma Foundation (ONF) studentships and fellowships in terms of capacity building in the neurotrauma field in Ontario. METHOD: An online, cross sectional survey amongst past recipients of studentships and fellowships that terminated prior to July 2005. Explicit data were collected on various aspects of career development including current activity, awards and publications. RESULTS: Thirty-six out of 42 (86%) eligible past trainees responded; 12 (33%) were Masters students, 12 (33%) were PhD students and 12 (33%) were Post-Doctoral students. A majority of the recipients (61%) are currently involved in neurotrauma-related activities (clinical, research and teaching) in more than 20% of their time, with no substantial differences between the degree groups. Half the recipients are currently involved in neurotrauma-related research in more than 20% of their time. The awardees published 1.5 peer-review manuscripts/person-year and received multiple awards. A high majority of our recipients (86%) feel that the ONF award had a substantial impact on their career. CONCLUSIONS: A high proportion of past award recipients remain involved in neurotrauma activities, especially in research. These results may lead to a cautious conclusion of the positive impact of the ONF studentships and fellowships on neurotrauma capacity building. These results should be considered in strategic planning of funding agencies similar to ONF. PMID- 18380284 TI - Left eye proptosis in an 11-year-old child. PMID- 18380285 TI - Metabolic myopathies discovered during investigations of statin myopathy. PMID- 18380286 TI - Glioblastoma in a patient with a hereditary cancer syndrome. PMID- 18380287 TI - Spontaneous C1-2 CSF leak treated with high cervical epidural blood patch. PMID- 18380288 TI - Anterior corpectomy approach for removal of a cervical intradural schwannoma. PMID- 18380289 TI - Nodular fasciitis presenting in the obturator nerve and gracilis muscle. PMID- 18380290 TI - Cryptogenic methicillin-resistant Staphylococcus aureus brain abscess. PMID- 18380291 TI - An unusual case of stroke and fevers in a traveler returning from Arizona. PMID- 18380292 TI - Hand dermatitis: a review of clinical features, diagnosis, and management. AB - Hand dermatitis is a common skin condition among all ages and both genders. Diagnosis, treatment, lifestyle changes, and an overall understanding of this condition are important in controlling severity and outcomes. PMID- 18380293 TI - Dermatology and skin disease in the American Civil War. AB - The Civil War happened at the end of the medical dark ages or, conversely, at the beginning of the modern medical era. The story of how physicians and nurses of the time approached a number of cutaneous diseases of importance in the military context is related. Entities discussed include the army itch/camp itch phenomenon, sexually transmitted diseases, scurvy and nutritional disorders, smallpox and spurious vaccination, and hospital gangrene. PMID- 18380294 TI - Skin infections in athletes. AB - Athletes represent a unique group of dermatologic patients. Intense and close contact with others' skin primarily places the athlete at risk for infection. Failure to acknowledge the role of sports participation in the genesis of skin infections leads to delayed diagnoses, inappropriate therapy, and subsequent epidemics. Simple prevention measures curb epidemics and allow teams to practice and compete without disruption. PMID- 18380295 TI - What's your assessment? Varicella. PMID- 18380296 TI - Issues related to participation in a clinical trial. AB - While not everyone can or should participate in a clinical trial, these treatment plans should be discussed with appropriate patients and their family members. A clinical trial is a precise, exact written protocol that allows patients in multiple health care settings to have access to these newer modalities. PMID- 18380297 TI - Viral infections. PMID- 18380298 TI - Trichotillomania. PMID- 18380299 TI - Living with psoriasis. PMID- 18380300 TI - ["Nothing gets done without the health professionals". Interview by Yasmina Ouharzoune with Roselyne Bachelot, minister of health, youth and sports]. PMID- 18380304 TI - [Improving care of chronic wounds]. PMID- 18380305 TI - [Hygiene and prevention of infectious risks in community health professionals]. PMID- 18380306 TI - ["There are things to be done in favor of nurses and access to care". Interview by Serge Benaderette]. PMID- 18380307 TI - [The administration of goserelin and the nursing role during injections visits]. PMID- 18380308 TI - [Taking care of leukemia patients]. PMID- 18380309 TI - [Epidemiology of leukemias]. PMID- 18380310 TI - [Hematopoiesis, a complex system]. PMID- 18380311 TI - [Different types of leukemias]. PMID- 18380312 TI - [Treatment of acute leukemias and chronic myeloid leukemias]. PMID- 18380313 TI - [Place and role of the nurse in the diagnosis disclosure in hematology]. PMID- 18380314 TI - [The hospital nurse and chemotherapy]. PMID- 18380315 TI - [Home and chemotherapy, for a better partnership]. PMID- 18380316 TI - [Bone marrow transplantation and nursing care]. PMID- 18380317 TI - [Psychological work after the diagnosis disclosure of leukemia]. PMID- 18380318 TI - [Emergency admission in hematology, the nursing team organizes itself]. PMID- 18380319 TI - [Evolution of treatments, what upheavals?]. PMID- 18380320 TI - [2/10 Carbohydrates, water and fiber]. PMID- 18380321 TI - [2/11 The patient with a total knee prosthesis]. PMID- 18380322 TI - [Investigation of radiation-induced "bystaner effect" using model of adaptive response in mixed lymphocyte culture from humans of different gender]. AB - The novel method for the investigation of radiation-induced "bystander effect" has been tested on the model of mixed lymphocyte culture from humans of different gender. The "bystander effect" was estimated by the ability of nonirradiated female/male cells to develop an adaptive response in mixed culture with irradiated at the dose 0.05 Gy of X-rays G0 lymphocytes of opposite gender. The preliminary results indicate that both irradiated lymphocytes and non-irradiated but neighbouring with pre-exposed cells are less susceptible to the genetic damages manifested as chromosome aberrations induced in G1 lymphocytes by a subsequent high dose of X-ray (1.0 Gy). Direct adaptive response as well as indirect one were expressed more obvious in female lymphocytes. PMID- 18380323 TI - [Stress signaling between human lymphocytes after induction of bystander effect by exposure to ionizing radiation in adaptive doses]. AB - We previously reported that the consequence of human lymphocytes irradiation by the adaptive doses (X-rays, 10 cGy) was a transposition of the homologous chromosome loci in the cell nucleus (FISH method); this phenomenon was mediated by the increase of nucleolus activity. They both are transmited to non-irradiated cells by the bystander effect (BE). We shown that the reaction of stress signaling is induced by the DNA fragments of irradiated lymphocytes. The study shows that after the inhibition of caspase 3 activity in irradiating lymphocytes or the blockade TLR9 in bystander cells the transposition was not observed. A signaling way of BE from irradiated lymphocytes apoptosis to bystander cells receptors is discussing. PMID- 18380324 TI - [An individual variability of the adaptive response to irradiation in human cells. Approach to its determination]. AB - On human blood lymphoxytes with micronuclei (MN) assay and cytokinetic cytochalasin block and analysis of chromosome aberrations the change of cell population composition, adaptive response (AR) and phenomenon of enhanced radiosensitivity after low dose (5 cGy) and challenge doses (1.0 Gy) have been studied. Irradiation have been carried out in G1 and G2 phases of cell cycle (24 h and 48 h after PHA stimulation). Fixation of cells have been conducted after 50 h (2 h after demecolcin adding) and 72 h (24 h after cytochalasin adding) chromosome and MN assay. Evaluation criteria were the frequency of binucleated cells with MN on 1000 binucleated cells and the frequency of cells with chromatid aberration on 100 metaphases. It was shown that cell population constitution change, AR occurring depended on the individual peculiarity. The evaluation of AR presence by the indexes of bimucleated cells with MN frequency and cells with chromatid aberrations don't coincide (coincidence is observed in 3 cases from 15). It is supposed that in G2 phase after irradiation in challenge dose the MN assay and metaphase analysis can register different cells (24 h and 2 h after mitotic block). The cell population constitution change can probably influence on the AR evaluation but in isn't the AR mechanism. The main mechanism of AR forming * the protection from the damages by different ways. AR depends on many factors, individual peculiarities observes by the use of definite evaluation criteria, in individuals with definite genetic constitution. Perhaps these considerations permit to discuss the problem of AR universality. PMID- 18380325 TI - [Effect of temperature on the development of chromosome aberrations in human blood lymphocytes irradiated in pulse and continuous modes at BARS-6 reactor]. AB - Summarized results of 5 repeated experiments of comparative study of radiation effects of the pulse reactor BARS-6 either in a single pulse or a continuos irradiation mode on human lymphocytes are presented. Higher efficiency (30-40%) of continuous irradiation (exposure duration 1 h) rather than pulse irradiation with ultra-high dose rates (1-2.5) x 10(6) Gy/min (pulse duration 65 micros) was confirmed. The efficiency ratio did not depend on the temperature, 20 degrees C or 0 degrees C, during the exposure. Cell repair system and chromatin conformation influence on the results obtained is discussed. PMID- 18380326 TI - [Ionizing radiation can activate the insertion of mitochondrial DNA fragments in the nuclear genome]. AB - In analytical review is considered the possibility of the insertion of mitochondrial DNA (mtDNA) fragments into the nuclear genome of cells, exposed ionizing radiation (IR). Many studies show that integration fragment mtDNA in nuclear genome, as well as its fastening as NUMT-pseudogenes, proceed at ancient periods of the evolutions not only, but also at more late periods. The number of the investigations shows that under influence endogenous reactive oxygen species, chemical agent, UV-light and IR mtDNA is damaged with greater frequency, than nucleus DNA. Furthermore, the repair systems in mitochondria are low efficiency. In irradiated by IR cells mtDNA fragments can transition from the mitochondria to the cytoplasm. The binding of mtDNA fragment to a complex with proteins provides them the protection from nuclease destroying. Possibly, at such safe condition they and are carried to nucleus. At inductions of DNA double-strand breaks (under the action of IR and activated their reparation) mtDNA fragments may be inserted to nuclear genome. Such integration of mtDNA to nuclear genome, with shaping NUMT pseudogenes de novo, may be proceed in irradiated cells in the course of the reparations DNA double-strand breaks by the nonhomologous end-joining pathway. These insertions of mtDNA can cardinally change the structure of nuclear genomes in area of their introduction and render the essential influence upon the realization of genetic information. Available information in literature also allows to suppose that integration mtDNA in nuclear genome can proceed and at raised genomic instability observed in cells at post radiation period. It in equal extent pertains and to malignant cells with raised by instability mitochondrial and nuclear genomes. As the most efficient agent, initiating insertion fragment mtDNA in nuclear genome, is considered ionizing radiation. PMID- 18380327 TI - [Experimental study of the radioprotective action of melanin on the somatic development during irradiation in the antenatal period of ontogenesis]. AB - Daily introduction per os of the exogenous melanin in a doze of weight of 10 mg/kg pregnant female rats Wistar eliminated the functional deficiency of somatic development revealed at posterity at chronic gamma-irradiation in a doze 1.00 1.25 Gy for all period of pregnancy. The irradiation or introduction melanin antenatal to a phase ontogenesis resulted in stimulation of the immune answer, which was determined at offspring on 3rd week after birth. On the basis of the received data it is concluded presence radioembryoprotective actions melanin in the relation embryotoxic effects of small dozes of ionizing radiation and its participation in regulation immunogenes. PMID- 18380328 TI - [Experimental study of the neuroprotective properties of the melanin in embryos irradiated during antenatal development]. AB - Daily introduction per os of the exogenous melanin in a doze of weight of 10 mg/kg pregnant female rats of line Wistar on a background continuous irradiations (dose rate of 2.6 mGy/h within 20 days of pregnancy) eliminated deficiency cognitive functions at posterity. On the basis of the received data it is concluded presence radioembryoprotective actions of melanin in the relation neuro embryotoxic effects of small dozes ionizing radiation. Taking into account small toxicity of melanin, the preparation can be perspective for practical application. PMID- 18380329 TI - [Serum TNFalpha and its receptors in professionals after prolonged exposure to radiation in remote period]. AB - The goal of this study was the detection of serum TNFalpha concentration and soluble receptors (sTNFRI and sTNFRII) in PA Mayak professionals in long-term after the prolonged radiation exposure, subject to cumulative dose. TNFalpha, sTNFRI, and sTNFRII concentration were investigated by ELISA in serum of 59 professionals. Cumulative external doses were within 0.21-7.72 Gy. 239Pu body burden was less than 1.48 kBk. Direct significant correlation was detected between TNFalpha concentration and external dose (0.21-3.(00) Gy; r = 0.41, p = 0.009, n = 40). Serum sTNFRI concentration rose significantly with increasing of cumulative dose. Significant increase of persons number with deviations of sTNFRI concentration from standards in groups with dose higher 1.00 Gy under comparison with control was discovered. The correlation was detected between external dose and serum sTNFRI concentration in long-term after radiation exposure (r = 0.36, p < 0.005). Significant positive correlation was detected between sTNFRI, and sTNFRII under increasing of cumulative external dose (r = 0.60, p = 0.0000004). PMID- 18380330 TI - [Early leukemia effect during chronic exposure to radiation with high doses]. AB - The preliminary results of the analysis of leukemia morbidity in the sub-cohort of workers from PA "Mayak" exposed with high (more than 4Gy) doses during relatively short time range (few years) have been obtained in terms of materials from the medical-dosimetry register (SUrIBPh). The earlier dynamics of the leukemia morbidity implementation (2-5 years after the beginning of exposure) was established for this sub-cohort, in contrast to that predicted on the base of examination of the cohort of atomic bombardment victims from Japan cities (LSS). The "'early" leukemia effect is connected with intensive cell death and has a threshold nature. It could be supposed, that intensification of born-marrow hematopoetics restricts a potential of reparation processes and leads to earlier (in contrast with that observed in the LSS cohort) implementation of carcinogenetic effect. Using propositions developed to describe the process of the creation of consecutive specific stable mutation in the target cells, the options is proposed of multistage model, which allows the prediction of post radiation dynamics of leukemia morbidity intensity. Both data from LSS(DS86) and from the register for workers from PA "Mayak" were used to asses the model parameters. The satisfactory agreement is illustrated between the observed dynamics of leukemia morbidity and the model calculations. PMID- 18380331 TI - [Electrophoretic and immunochemical research of rat urine proteins in dynamics after intravenous injection of thorium dioxide (thorotrast)]. AB - Rats were treated with a single intravenous injection of thorotrast (thorium dioxide)--the source of alpha-rays. Dynamic investigation of urine protens of rats by methods of electrophoresis and immunoelectrophoresis was carried out during 22 months after thorotrast injection. Already the month after drug injection the selectivity of tubular reabsorbtion was disturbed. Three months after thorotrast injection the content of urinal proteins of tissue (in particular renal) origin was decreased. Finally the selectivity of renal filtration of proteins was damaged 4-6 months after thorotrast introduction. Serum proteins which were absent in normal urine (for example transferrin and lipoproteins) appeared in urine of affected rats. The urine proteins of serum origin were less degraded than those in normal urine. The alterations of glomerular filtration was increased up to 20-22 months when the spectrum of urine proteins became similar to the spectrum of serum proteins. The death of treated rats was occurred in this period. Thus the monitoring of urine proteins of rats treated with alpha-ray producing preparation throtrast allows to register the successive alterations of reabsorbtion, excretion and filtration functions of kidney. PMID- 18380332 TI - [Free-radical mechanism of chitosan radiation degradation and problems of the chemical antiradiation protection]. AB - An analysis of published data on the radiolytic properties of chitosan and of some modelling its fragments substances was performed. The main process of radiation chitosan degradation is connected with the formation and conversion of free radicals. The mechanisms of the primary radicals C2, C1 and C3 (of the NH2 and H removal radicals) conversion up to the formation of some terminal products of chitosan radiolysis were offered in the form of the generalized schemes. Problem of the DNA and membrane protection in irradiated cell is discussed. PMID- 18380333 TI - Problems in assessment of risks from exposures to microwaves of mobile communication. AB - Since pioneering investigations published in the beginning of 1970th, various biological responses to non-thermal (NT) microwaves (MW), including adverse health effects, have been described by many research groups all over the world. There is strong evidence that the NT MW biological effects depend on several physical parameters and biological variables, which must be controlled in replication studies. Apart from the fundamental importance, the development of comprehensive mechanisms for the NT MW effects is socially important. The effects of MW of mobile communications are of major concern because of the increased exposure in many countries. It has been shown that adverse effects of NT MW from GSM/UMTS mobile phones on human lymphocytes from healthy and hypersensitive to EMF persons depend on carrier frequency and modulation. Further investigations with human primary cells, animals and volunteers are needed to elucidate possible adverse effects of MW signals that are used in wireless communication. Identification of those types and frequency channels/bands for mobile communication, which do not affect human cells, is urgently needed as the high priority task for the development of safe mobile communication. Numerous data on the NT MW effects clearly indicate that the SAR-concept alone cannot underlie the safety guidelines for chronic exposures to MW from mobile communication and other approaches are needed. However, there is not enough research information to set exposure MW standards. Various genetic and epigenetic effects of signals used in mobile communication should be studied. It has been shown that NT MW affect cells of various types including stem cells and reproductive organs. Stem cells represent especially important cellular model because recent data suggest that different cancer types, including leukemia, have a fundamentally common basis that is grounded on epigenetic changes in stem cells. PMID- 18380334 TI - [The results of computerization of studies of ecological consequences of the accident on the Chernobyl Nuclear Power Plant for terrestrial ecosystems]. AB - In the paper some results of application of information and calculation technologies in researches of ecological consequences of accident on Chernobyl NPP are brought. The effectiveness of a computerization of investigations is scored. Technical and information component isselected. The singularities of application of the methodology of computerization in radioecological researches are considered. The special attention is given to integration of knowledge accumulated in the form of information materials, databases, mathematical models. The browse of the series of radioecological information and of informational prognostic systems constructed from the moment of accident is made. As an example of successful usage of a computerization in radioecological researches provided by small scientific collectives experience of MSU division of radioecology and ecotoxicology are considered. PMID- 18380335 TI - [Age specific formation of doses in children exposed to radioactive caesium following the accident in the Chernobyl Nuclear Power Plant]. AB - Individual measurement data on radiological surveillance of people residing in the territories most radioactively contaminated following the Chernobyl accident are analyzed to provide a comparison of doses from caesium radionuclides to children and adults. The external doses to children does not exceed those to adult inhabitants of the same settlement. By the results of surveillance in 1986 more than 80 thousand inhabitants of the western areas of Bryansk region, a specific activity of caesium radionuclides and corresponding internal radiation dose rate in a number of settlements appeared at children age under 3 years old exceeding the appropriate parameters for adult inhabitants. Among persons evacuated from the contaminated areas, such excess in day of departure from an area averaged 3.8 for surveyed pairs the child-parent. PMID- 18380336 TI - [The assessment of 137Cs accumulation by pine tree plantations in the closest radioactive fallouts originated from the Chernobyl Nuclear Power Plant]. AB - The assessment of 137Cs accumulation by wood is made by using a chamber model of radionuclide behavior. Maximum of 137Cs specific activity in wany wood was in the midst of 1990s, with subsequent decrease in activity and half-time up to 22 to 25 years. PMID- 18380337 TI - Nuclear organization and splicing control. AB - Although major splicing regulatory mechanisms rely on the presence of cis-acting sequence elements in the precursor messenger RNA (pre-mRNA) to which specific protein and factors bind, splice choices are also influenced by transcription kinetics, promoter-dependent loading of RNA-binding proteins and nucleo cytoplasmic distribution of splicing regulators. Within the highly crowded eukaryotic nucleus, molecular machines required for gene expression create specialized microenvironments that favor some interactions while repressing others. Genes located far apart in a chromosome or even in different chromosomes come together in the nucleus for coordinated transcription and splicing. Emerging tools to dissect gene expression pathways in living cells promise to provide more detailed insight as to how spatial confinement contributes to splicing control. PMID- 18380338 TI - Spliceosome assembly and composition. AB - Cells control alternative splicing by modulating assembly of the pre-mRNA splicing machinery at competing splice sites. Therefore, a working knowledge of spliceosome assembly is essential for understanding how alternative splice site choices are achieved. In this chapter, we review spliceosome assembly with particular emphasis on the known steps and factors subject to regulation during alternative splice site selection in mammalian cells. We also review recent advances regarding similarities and differences between the in vivo and in vitro assembly pathways, as well as proofreading mechanisms contributing to the fidelity of splice site selection. PMID- 18380339 TI - Relating alternative splicing to proteome complexity and genome evolution. AB - Prior to genomics, studies of alternative splicing primarily focused on the function and mechanism of alternative splicing in individual genes and exons. This has changed dramatically since the late 1990s. High-throughput genomics technologies, such as EST sequencing and microarrays designed to detect changes in splicing, led to genome-wide discoveries and quantification of alternative splicing in a wide range of species from human to Arabidopsis. Consensus estimates of AS frequency in the human genome grew from less than 5% in mid-1990s to as high as 60-74% now. The rapid growth in sequence and microarray data for alternative splicing has made it possible to look into the global impact of alternative splicing on protein function and evolution of genomes. In this chapter, we review recent research on alternative splicing's impact on proteomic complexity and its role in genome evolution. PMID- 18380341 TI - Technologies for the global discovery and analysis of alternative splicing. AB - During the past approximately 20 years, studies on alternative splicing (AS) have largely been directed at the identification and characterization of factors and mecha nisms responsible for the control of splice site selection, using model substrates and on a case by case basis. These studies have provided a wealth of information on the factors and interactions that control formation of the spliceosome. However, relatively little is known about the global regulatory properties of AS. Important questions that need to be addressed are: which exons are alternatively spliced and under which cellular contexts, what are the functional roles of AS events in different cellular contexts, and how are AS events controlled and coordinated with each other and with other levels of gene regulation to achieve cell- and development-specific functions. During the past several years, new technologies and experimental strategies have provided insight into these questions. For example, custom microarrays and data analysis tools are playing a prominent role in the discovery and analysis of splicing regulation. Moreover, several non-microarray-based technologies are emerging that will likely further fuel progress in this area. This review focuses on recent advances made in the development and application of high-throughput methods to study AS. PMID- 18380340 TI - Complex alternative splicing. AB - Alternative splicing is a powerful means of controlling gene expression and increasing protein diversity. Most genes express a limited number of mRNA isoforms, but there are several examples of genes that use alternative splicing to generate hundreds, thousands and even tens of thousands of isoforms. Collectively such genes are considered to undergo complex alternative splicing. The best example is the Drosophila Down syndrome cell adhesion molecule (Dscam) gene, which can generate 38,016 isoforms by the alternative splicing of 95 variable exons. In this review, we will describe several genes that use complex alternative splicing to generate large repertoires of mRNAs and what is known about the mechanisms by which they do so. PMID- 18380342 TI - Searching for splicing motifs. AB - Intron removal during pre-mRNA splicing in higher eukaryotes requires the accurate identification of the two splice sites at the ends of the exons, or exon definition. The sequences constituting the splice sites provide insufficient information to distinguish true splice sites from the greater number of false splice sites that populate transcripts. Additional information used for exon recognition resides in a large number of positively or negatively acting elements that lie both within exons and in the adjacent introns. The identification of such sequence motifs has progressed rapidly in recent years, such that extensive lists are now available for exonic splicing enhancers and exonic splicing silencers. These motifs have been identified both by empirical experiments and by computational predictions, the validity of the latter being confirmed by experimental verification. Molecular searches have been carried out either by the selection of sequences that bind to splicing factors, or enhance or silence splicing in vitro or in vivo. Computational methods have focused on sequences of 6 or 8 nucleotides that are over- or under-represented in exons, compared to introns or transcripts that do not undergo splicing. These various methods have sought to provide global definitions of motifs, yet the motifs are distinctive to the method used for identification and display little overlap. Astonishingly, at least three-quarters of a typical mRNA would be comprised of these motifs. A present challenge lies in understanding how the cell integrates this surfeit of information to generate what is usually a binary splicing decision. PMID- 18380343 TI - SR proteins and related factors in alternative splicing. AB - SR proteins are a family of RNA binding proteins that contain a signature RS domain enriched with serine/arginine repeats. The RS domain is also found in many other proteins, which are collectively referred to as SR-related proteins. Several prototypical SR proteins are essential splicing factors, but the majority of RS domain-containing factors are characterized by their ability to alter splice site selection in vitro or in transfected cells. SR proteins and SR related proteins are generally believed to modulate splice site selection via RNA recognition motif-mediated binding to exonic splicing enhancers and RS domain mediated protein-protein and protein-RNA interactions during spliceosome assembly. However, the biological function of individual RS domain-containing splicing regulators is complex because of redundant as well as competitive functions, context-dependent effects and regulation by cotranscriptional and post translational events. This chapter will focus on our current mechanistic understanding of alternative splicing regulation by SR proteins and SR-related proteins and will discuss some of the questions that remain to be addressed in future research. PMID- 18380344 TI - hnRNP proteins and splicing control. AB - Proteins of the heterogeneous nuclear ribonucleoparticles (hnRNP) family form a structurally diverse group of RNA binding proteins implicated in various functions in metazoans. Here we discuss recent advances supporting a role for these proteins in precursor-messenger RNA (pre-mRNA) splicing. Heterogeneous nuclear RNP proteins can repress splicing by directly antagonizing the recognition of splice sites, or can interfere with the binding of proteins bound to enhancers. Recently, hnRNP proteins have been shown to hinder communication between factors bound to different splice sites. Conversely, several reports have described a positive role for some hnRNP proteins in pre-mRNA splicing. Moreover, cooperative interactions between bound hnRNP proteins may encourage splicing between specific pairs of splice sites while simultaneously hampering other combinations. Thus, hnRNP proteins utilize a variety of strategies to control splice site selection in a manner that is important for both alternative and constitutive pre-mRNA splicing. PMID- 18380345 TI - Functional and mechanistic insights from genome-wide studies of splicing regulation in the brain. AB - We review here results arising from the systematic functional analysis of Nova, a neuron-specific RNA binding protein targeted in an autoimmune neurological disorder associated with cancer. We have developed a combination of biochemical, genetic and bioinformatic methods to generate a global understanding of Nova's role as a splicing regulator. Genome-wide identification and validation of Nova target RNAs has yielded unexpected insights into the protein's mechanism of action and into the functionally coherent role of Nova in the biology of the neuronal synapse. These studies provide us with a platform for understanding the role of RNA binding proteins in tissue-specific splicing regulation and in disease. PMID- 18380346 TI - Regulation of alternative splicing by signal transduction pathways. AB - Alternative splicing is now recognized as a ubiquitous mechanism for controlling gene expression in a tissue-specific manner. A growing body of work from the past few years as begun to also highlight the existence of networks of signal responsive alternative splicing in a variety of cell types. While the mechanisms by which signal transduction pathways influence the splicing machinery are relatively poorly understood, a few themes have begun to emerge for how extracellular stimuli can be communicated to specific RNA-binding proteins that control splice site selection by the spliceosome. This chapter describes our current understanding of signal-induced alternative splicing with an emphasis on these emerging themes and the likely directions for future research. PMID- 18380347 TI - Coupling transcription and alternative splicing. AB - Alternative splicing regulation not only depends on the interaction of splicing factors with splicing enhancers and silencers in the pre-mRNA, but also on the coupling between transcription and splicing. This coupling is possible because splicing is often cotranscriptional and promoter identity and occupation may affect alternative splicing. We discuss here the different mechanisms by which transcription regulates alternative splicing. These include the recruitment of splicing factors to the transcribing polymerase and "kinetic coupling", which involves changes in the rate of transcriptional elongation that in turn affect the timing in which splice sites are presented to the splicing machinery. The recruitment mechanism may depend on the particular features of the carboxyl terminal domain of RNA polymerase II, whereas kinetic coupling seems to be linked to how changes in chromatin structure and other factors affect transcription elongation. PMID- 18380348 TI - The coupling of alternative splicing and nonsense-mediated mRNA decay. AB - Most human genes exhibit alternative splicing, but not all alternatively spliced transcripts produce functional proteins. Computational and experimental results indicate that a substantial fraction of alternative splicing events in humans result in mRNA isoforms that harbor a premature termination codon (PTC). These transcripts are predicted to be degraded by the nonsense-mediated mRNA decay (NMD) pathway. One explanation for the abundance of PTC-containing isoforms is that they represent splicing errors that are identified and degraded by the NMD pathway. Another potential explanation for this startling observation is that cells may link alternative splicing and NMD to regulate the abundance of mRNA transcripts. This mechanism, which we call "Regulated Unproductive Splicing and Translation" (RUST), has been experimentally shown to regulate expression of a wide variety of genes in many organisms from yeast to human. It is frequently employed for autoregulation of proteins that affect the splicing process itself. Thus, alternative splicing and NMD act together to play an important role in regulating gene expression. PMID- 18380349 TI - Alternative splicing in disease. AB - Alternative splicing is a major source of diversity in the human proteome. The regulation of alternative splicingmodulates the composition of this diversity to fulfill the physiological requirements of a cell. When control of alternative splicing is disrupted, the result can be a failure to meet cellular and tissue requirements resulting in dysfunction and disease. There are several well characterized examples in which disruption of alternative splicing is a cause of disease. Investigations into how the mis-regulation of alternative splicing causes disease complements investigations of normal regulatory processes and enhances our understanding of regulatory mechanisms in general Ultimately, an understanding of how alternative splicing is altered in disease will facilitate strategies directed at reversing or circumventing mis-regulated splicing events. PMID- 18380350 TI - [Gnothi seauton--know thyself! Do you want to survey your genome?]. PMID- 18380351 TI - [Food additives can increase hyperactivity in children. Results from a British study confirm the connection]. PMID- 18380352 TI - [Interferon-alpha can improve the prognosis in high-risk melanoma. Combination of surgery, cytostatics and natural IFN-alpha doubled the survival rate]. PMID- 18380353 TI - [National medical indications. The key to evidence-based, just and accessible care]. PMID- 18380354 TI - [Hospital alcohol tests not completely easy to use for legal purposes. Conversion of ethanol levels in plasma or serum to permillage level in blood]. PMID- 18380355 TI - [Internship should give competence in medical science. The Surgical Association's curriculum proposal for all interns]. PMID- 18380356 TI - [Do Cardizem and Waran interact?]. PMID- 18380357 TI - [Organized improvement of cervix cancer prevention]. PMID- 18380358 TI - [Information about prenatal diagnosis doesn't fulfil comprehensiveness requirements]. PMID- 18380359 TI - [Combined nuchal translucency and biochemistry screening--unanswered questions and moral conflicts]. PMID- 18380360 TI - [Babel's House revisited...]. PMID- 18380361 TI - [One-eyedness on who are suitable as physician]. PMID- 18380362 TI - [Low prescription rates of generic lamotrigine and gabapentine]. PMID- 18380363 TI - [View on patients with pain needs to be changed]. PMID- 18380364 TI - [The anatomy of the soul--does it exist?]. PMID- 18380370 TI - [The transmission of knowledge in mental health]. PMID- 18380371 TI - [Detours and new contours in psychiatric nursing education]. PMID- 18380372 TI - [Training and consolidation of knowledge in the field]. PMID- 18380373 TI - [Tutorial, institutional steps and pedagogic project]. PMID- 18380374 TI - [The tutorial, original space for clinical thinking]. PMID- 18380375 TI - [Consolidation of knowledge in psychiatry and clinical nursing]. PMID- 18380376 TI - [Merlin, the goblin and the young traveller]. PMID- 18380377 TI - [Putting practice at the heart of knowledge consolidation]. PMID- 18380378 TI - [Psychiatric food]. PMID- 18380379 TI - [Complaints of back pain in childhood: find curable causes]. AB - Three children, a 13-year-old boy and a 3-year-old and 6-year-old girl, were presented to the hospital with back pain, caused by Scheuermann's disease, spondylodiscitis and sickle cell disease, respectively. The boy with Scheuermann's disease received exercise therapy, the spondylodiscitis was treated with antibiotic therapy and the girl with sickle cell disease was given hyperhydration and folic acid. Although back pain is a common problem in children and teenagers, it is infrequently reported in the clinic. In contrast to back pain in adults, the same complaint in childhood is more often caused by a serious disorder which should be treated. Various causes of back pain in children can be distinguished: mechanical problems, infections of the lumbar spine, neoplasia, inflammation, and other causes, such as sickle cell disease. A child or adolescent presenting to the clinic with complaints of back pain deserves a careful detailed evaluation of the history, appropriate physical examination and additional investigation. Alarm symptoms are an increase in back pain, age below 4 years, pain during the night, restriction in function, systemic complaints or neurological deficits. PMID- 18380380 TI - [Guideline for imminent preterm birth: insufficient adherence and implementation]. AB - The guideline for referral to perinatology centres in cases of imminent preterm birth at 24-26 weeks gestation, is poorly adhered to by Dutch gynaecologists. Unfortunately, the guideline can be interpreted in various ways and the reasons for non-adherence remain unclear. In addition, no measures were taken to implement the guideline when it was published. This means that the usefulness of the finding that the guideline is poorly adhered to is limited. PMID- 18380381 TI - [Pharmacological treatment of Bell's palsy: favourable effects of prednisolone based therapy now demonstrated]. AB - Bell's palsy accounts for two-thirds ofall acute facial palsies. Presumed reactivation of the herpes simplex virus and concurrent swelling of the facial nerve prompted the use of antivirals in combination with corticosteroids, although evidence supporting the effectiveness of this approach was weak. A recently published randomized placebo-controlled clinical trial assessed the effectiveness of adding valacyclovir to prednisolone; another larger primary-care based study compared treatment with prednisolone, acyclovir or both with placebo. In patients with severe or complete facial palsy, the addition of valacyclovir improved the chance of complete recovery, but as this study was single-blinded, results should be interpreted with caution. Early treatment with prednisolone (25 mg twice daily for to days) significantly improved the chance of complete recovery at 3 and 9 months. Acyclovir, given alone or in addition to prednisolone, did not show any benefit. PMID- 18380382 TI - [The value of positron emission tomography in the diagnosis and treatment of oesophageal cancer]. AB - Fludeoxyglucose positron emission tomography (FDG-PET) is a noninvasive imaging technique that applies the glucose metabolism to visualise the metabolic activity ofa tumour. FDG-PET might improve the selection of potentially curable patients with oesophageal cancer in addition to state-of-the-art conventional work-up (e.g. endoscopic ultrasonography and spiral CT). The additional value however is only 4% for all patients, and 7% in patients with stage III-IV disease. Moreover, the additional costs of FDG-PET are not compensated by the cost reduction ofprevented surgery. To improve the outcome of patients with oesophageal cancer the value ofneoadjuvant chemo- and/or radiotherapy is being investigated. FDG-PET seems to be a promising tool for the early assessment of response to neoadjuvant therapy. In case of non-response the ineffective neoadjuvant therapy can be stopped without further delaying appropriate surgery. FDG-PET might be able to improve the prediction of prognosis, in addition to commonly used histopathological factors. PMID- 18380383 TI - [Angiogenesis inhibitors for the systemic treatment of metastatic renal cell carcinoma: sunitinib, sorafenib, bevacizumab and temsirolimus]. AB - Treatment of patients with metastatic renal cell carcinoma is evolving rapidly due to the advent of novel targeted therapies. Improved knowledge of the underlying pathogenesis has led to the development of drugs that modulate the dominant signal transduction pathways for this disease, which results in inhibition of angiogenesis. Recent evidence indicates that the receptor tyrosine kinase inhibitor sunitinib prolongs progression-free survival compared with interferon-alpha, especially in patients with intermediate risk. Immunotherapy with interferon-alpha or high-dose interleukin-2 should still be considered for low-risk patients, particularly those with clear-cell tumours and metastases of the lung only. In patients who fail treatment with interferon-alpha, sorafenib has been shown to improve progression-free survival. High-risk patients may benefit from treatment with temsirolimus, which inhibits mammalian target of rapamycin (mTOR) kinase activity and has shown to improve overall survival. These angiogenesis inhibitors did not receive mention in the recently published guideline 'Renal cell carcinoma'. PMID- 18380384 TI - [Guideline 'Renal cell carcinoma']. AB - Each year, more than 1500 new cases of renal cell carcinoma are diagnosed in the Netherlands, and approximately 850 patients die due to this disease. The guideline 'Renal cell carcinoma' contains clinical practice recommendations on the diagnosis (imaging, pathological assessment, histopathological classification) and treatment (surgery, chemo-, immuno-, and radiotherapy) of renal cell carcinoma. For diagnostic imaging, chest and abdominal CT is recommended. Scintigraphy is not recommended. The term 'Grawitz tumour' is obsolete and should be replaced by 'renal cell carcinoma' with histological subtype specification according to the 2004 WHO classification. Laparoscopic radical nephrectomy is as effective as open surgery for localised tumours (T1 and T2) and possibly also for T3 tumours. The laparoscopic approach is associated with less morbidity due to the less invasive nature of this technique. This operation requires experience. In partial nephrectomy, a small margin of healthy tissue is sufficient. Frozen section examination of the resection edges does not appear to be required. In patients with metastatic renal cell carcinoma who are eligible for immunotherapy, removal of the tumour prolongs survival. Metastasectomy prolongs survival in patients with a solitary metastasis. Most currently available cytotoxic agents are ineffective against renal cell carcinoma. Interferon-alpha may have a role in the treatment of patients with renal cell carcinoma and favourable prognostic factors, given the survival advantage demonstrated with this agent in clinical trials. The guideline is available in English at www.oncoline.nl. PMID- 18380385 TI - [Diagnostic image (361). A boy with headache and vomiting 2 days after a head injury]. AB - An 11-year-old boy presented 2 days after a head injury with headache and vomiting. There were no neurological abnormalities. CT of the head demonstrated a large epidural haematoma and a cranial fracture. PMID- 18380386 TI - [At the limits of viability: Dutch referral policy for premature birth too reserved]. AB - OBJECTIVE: Evaluation of policy and treatment of deliveries at the limits of viability in the Netherlands and resulting survival figures. DESIGN: Cohort study. METHOD: Within the framework of the European 'Models of organising access to intensive care for very preterm births in Europe' (MOSAIC) study, data was collected on all 512 births in 2003 (terminations excluded) following 22-31 weeks gestation in the catchment areas of the perinatal centres in Nijmegen and Utrecht, the Netherlands. RESULTS: Gynaecologists and neonatologists practised a reserved policy for the active treatment of pregnancies under 25 weeks (5/77; 6%); all infants died. At 25 weeks, an active obstetric policy was used in one quarter of pregnancies, but none of the infants survived. Even at 26 weeks pregnancy, the obstetric policy was reserved and the mortality relatively high (9/31; 29%). From the neonatal deaths, 86 out of 92 (93%) were preceded by a decision either not to start or to discontinue treatment. CONCLUSION: Dutch obstetricians and neonatologists practised a reserved policy at the limits of neonatal viability. There is more need for active antenatal transfer to perinatal centres for those at the lower limit of neonatal viability to enable well balanced decisions to take place. The parents' wishes should always be taken into account. PMID- 18380387 TI - [Treatment of foetal supraventricular tachycardia with antiarrhythmic medication administered through the umbilical vein]. AB - Foetal supraventricular tachycardia (SVT) with hydrops foetalis is associated with a high morbidity and mortality rate. If SVT with hydrops foetalis persists despite transplacental therapy, direct foetal treatment can be initiated. One foetus was found to have SVT with hydrops foetalis during the 29th week of pregnancy, and the condition persisted despite transplacental treatment. Amiodarone was administered directly via the umbilical vein, and the SVT resolved. A second foetus was found to have SVT with hydrops foetalis during the 28th week of pregnancy. The condition persisted despite maternal antiarrhythmic medication. Direct treatment of the foetus with amiodarone was successful. Amiodarone is the treatment of choice for direct foetal therapy for SVT, and can be administered safely via the umbilical vein. Direct foetal therapy should be considered for the treatment of foetal SVT with hydrops foetalis that occurs in the first 31 weeks of pregnancy and persists despite adequate transplacental therapy. PMID- 18380388 TI - [Familial hemiplegic migraine resulting in recurrent coma]. AB - A 71-year-old woman presented with recurrent episodes of headache accompanied by hemihypoesthesia, fever, aphasia, reduced consciousness and worsening of pre existing ataxia. Brain imaging revealed atrophy of the cerebellum. The white cell count in the cerebrospinal fluid was slightly increased. The patient had a family history of migraine and cerebellar ataxia. DNA testing revealed a missense mutation in the CACNA1A gene, confirming the diagnosis of familial hemiplegic migraine. Familial hemiplegic migraine is a rare subtype of migraine with aura. It follows an autosomal dominant pattern of inheritance. Patients with familial hemiplegic migraine exhibit a wide spectrum of symptoms, which can hinder the diagnosis. If a patient presents with recurrent coma or encephalitis with or without cerebellar ataxia, familial hemiplegic migraine should be included in the differential diagnosis. PMID- 18380390 TI - [Overweight and mortality risk: no connection between overweight in middle and older age groups and increased mortality]. PMID- 18380391 TI - [Hypertension due to liquorice and liquorice tea consumption]. PMID- 18380392 TI - [The molecular epidemiology of hepatitis A in The Netherlands; the usefulness of typing isolated viral strains]. PMID- 18380393 TI - Syzygium cumini (L.) SKEELS (Myrtaceae) against diabetes--125 years of research. AB - Syzygium cumini (L.) SKEELS (syn. S. jambolanum DC, Eugenia jambolana LAM.) belongs to the medicinal plants most often recommended as an adjuvant therapy in type 2 diabetes. The plant was extensively studied during the last 125 years, approximately 100 case reports were reported already before the discovery of insulin. After the Second World War, research was concentrated on animal studies. Not all, but many of them reported some success in reducing type 2 diabetes symptoms. However, a state-of-the-art clinical study is still missing. In this review, historical literature dating back to the pre-insulin era was evaluated as were more recent in vitro-, animal-, and in vivo studies. Results were screened for information still useful today and compared to study results achieved in more recent decades. In view of the knowledge summarized here, a successful clinical study should use S. cumini seeds, seed kernels or fruit from India in fairly high doses. Reductions on blood sugar levels by about 30% seem reasonably to be expected. Adverse effects to be expected comprise gastrointestinal disturbances. PMID- 18380394 TI - Optimization of UDP-N-acetylmuramic acid synthesis. AB - UDP-N-acetylmuramic acid (UDP-MurNAc) is a substrate of MurC, an important enzyme in the intracellular pathway of bacterial peptidoglycan biosynthesis. Various approaches towards preparation of UDP-MurNAc have been published but these synthetic preparations were shown to include many problematic steps. An optimization study with the focus on muramyl phosphate and UMP-morpholidate coupling was performed, resulting in a synthetic procedure enabling robust and easily reproducible production on a multi-gram scale. PMID- 18380395 TI - Validated enantiospecific LC method for determination of (R)-enantiomer impurity in (S)-efavirenz. AB - A high-performance liquid chromatographic method was developed for separation of the enantiomers of efavirenz. The developed method was applied for the determination of (R)-enantiomer in (S)-efavirenz and satisfactory results were achieved. The base line separation with a resolution of more than 4.0 was achieved on Chiralcel OD (250 mm x 4.6 mm, 10 microm) column containing tris-(3,5 dimethylphenylcarbomate) as stationary phase. The mobile phase consists of n hexane: isopropyl alcohol (80:20 v/v) with 0.1% (v/v) of formic acid as additive. The flow rate was kept at 1.0 ml/min and the UV detection was monitored at 254 nm. The (R)-enantiomer was found linear over the range of 0.1 microg/ml--6 microg/ml. The limit of detection (LOD) was 0.03 microg/ml and the limit of quantification (LOQ) was 0.1 microg/ml (n=3. The precision of (R)-enantiomer at LOQ level was evaluated through six replicate injections and the RSD of the peak response was achieved as 1.34%. The results demonstrated that the developed LC method was simple, precise, robust and applicable for the purity determination of efavirenz. PMID- 18380396 TI - Determination of bergamottin in human plasma after grapefruit juice ingestion by an UPLC/MS/MS method. AB - Bergamottin was identified as a cause of pharmacokinetic interaction with grapefruit juice intake and as a physiologically active substance involved in lipolysis. However, the quantification method on concentrations of bergamottin in systemic circulation has not been well established. The aim of this study was to develop a simple, sensitive and high-throughput determination system for bergamottin in human plasma using an ultra performance liquid chromatography (UPLC)-MS-MS method. The UPLC system equipped with a UPLC BEH C18 column (2.1 x 50 mm, 1.7 microm) and an ESI prove was appropriate for detection of bergamottin. As a result, a primary product ion (m/z = 203) from precursor ion of bergamottin (m/z = 339) was observed. Plasma from a human volunteer who consumed grapefruit juice one hour before the time of blood sampling, was measured with the UPLC/MS/MS system. The determination of plasma-bergamottin was performed with the highest sensitivity presently available. In conclusion, we succeeded high performance bergamottin-determination in human plasma after grapefruit juice ingestion. The procedure can be usefulto clarify pharmacokinetic and pharmacodynamic characteristics of bergamottin. PMID- 18380397 TI - Deviations of drug solubility in water-cosolvent mixtures from the Jouyban-Acree model--effect of solute structure. AB - Deviations of the predicted solubilities using the Jouyban-Acree model from experimental data were correlated to the structural descritptors of the drugs computed by HyperChem software. The proposed models are able to predict the solubility in water-cosolvent mixtures and reduced the mean percentage deviations (MPD) of predicted solubilities from 24%, 48%, and 53% to 16%, 33% and 38%, respectively for water-propylene glycol, water-ethanol and water-polyethylene glycol 400 mixtures, with the overall improvement in prediction capability of the model being approximately 13%. PMID- 18380398 TI - Polymethacrylate based microparticulates of insulin for oral delivery, part II: solid state characterization. AB - The objectives of the present study were (A) to characterize insulin microparticles prepared by the coprecipitation process by size exclusion chromatography, differential scanning calorimetry, fourier-transform IR spectroscopy, and powder X-ray diffractometry, and (B) to study the solid state conformation of insulin before and after entrapment in the polymeric carrier. Microparticles were prepared by dissolving insulin in 0.01 N HCI and alcohol USP to get a final concentration of 32% v/v. Eudragit L100, a representative polymethyacrylate polymer, was then dissolved in this solution which was transferred to a beaker containing cold water with homogenization to obtain microparticulates. Insulin powder, microcapsules, and a physical mixture of insulin and Eudragit L100 were then analyzed by SEC-HPLC, DSC, FTIR, and XRD to observe changes in protein conformation as result of the manufacturing process. While DSC, XRD and FTIR results were of limited value due to limits of instrument sensitivity, size exclusion chromatography data indicated that higher order aggregates were not formed during microcapsule formation. It was concluded that formulating insulin into microparticles by the coprecipitation process is an attractive and stable method for protein delivery and might be suitable for oral delivery of insulin. PMID- 18380399 TI - Design and development of a mucoadhesive buccal film bearing progesterone. AB - The purpose of this research was to develop and evaluate mucoadhesive films for buccal administration of progesterone using film-forming and mucoadhesive polymers. Buccal films of chitosan bearing progesterone were prepared by solvent casting technique. The films have been evaluated in terms of film weight, thickness, density, surface pH, FT-IR, X-ray diffraction analysis, bioadhesion, swelling properties, in vitro drug release and in vivo studies. It was found that the film formulations of 2 cm2 size having weight in the range of 239 +/- 0.32 to 290 +/- 3.23 mg and film thickness were in the range of 0.49 +/- 0.21 to 0.60 +/- 0.26 mm. Density of the films was found to be 0.108 to 0.139 g/mL. Drug content was found to be uniform in a range of 9.21 +/- 0.051 to 9.67 +/- 0.086 mg/cm2 for formulation F1 to F4. Maximum bioadhesion force was recorded for PVP buccal films (formulation F2) i.e. 0.45 +/- 0.53 N as compared to other films. In vitro residence time was in range of 1.85 +/- 0.08 to 8.94 +/- 0.08 h. The drug release studies revealed that formulations follows non-fickian diffusion. In vivo residence time data confirmed that none of the polymers detached from the oral mucosa over the study period, which indicated that the bioadhesion values of all polymers were satisfactory to retain the film on the buccal mucosa. These mucoadhesive formulations could offer many advantages in comparison to traditional treatments and their efficacy as an effective contraception is assessed. PMID- 18380400 TI - Characterization of solid-state forms of mebendazole. AB - This study deals with the generation and characterization of various solid-state forms of mebendazole (MBZ), a benzimidazole antihelmentic. The drug was subjected to polymorphic screen using different solvents to explore the possibility of existence of different solid forms. Different reported polymorphic forms of MBZ, i.e. form A, B and C were found to be recrystallized from acetic acid:methanol mixture (1:1), ethyl acetate and methanol, respectively. N,N-Dimethyl acetamide (DMA) and N,N-dimethyl formamide (DMF) yielded two new solvates of MBZ. These solid-state forms were characterized by thermoanalytical (DSC, TGA, HSM), crystallographic (XRD), microscopic (optical, polarized), and spectroscopic (FTIR) techniques. Solubility studies were carried out for the solvates to identify the solubility advantage. Molecular modeling studies revealed moderately strong hydrogen bonding between the solvent molecules and MBZ. PMID- 18380401 TI - Drug interaction potentials among different brands of grapefruit juice. AB - The discrepancy of drug-interaction potential among different brands of grapefruit juice was estimated based on inhibition of CYP3A activity caused by furanocoumarin derivatives in the grapefruit juice. Heat treatment of the grapefruit juice at 95 degrees C for 1 h was utilized to degrade the furanocoumarins. Initial velocity of testosterone 6beta-oxidation using human liver microsomes was determined as an indicator of the CYP3A activities. Changes in the velocities of the reaction mixture were observed when 10% of each brand of untreated grapefruit juice or heat-treated grapefruit juice was added. The differences in the velocities between untreated and heat-treated grapefruit juice were defined as the potentials of furanocoumarin-caused CYP3A-inhibitions. PMID- 18380402 TI - Effects of LPS, lipid A and polysaccharide from adapted strains of Escherichia coli on human leucocyte activity. AB - Polysaccharide and lipid A are responsible for the wide-ranging pharmacological activity of bacterial lipopolysaccharides (LPS). The alterations in LPS structure result in various effects on different functions of the target cells. The effects of LPS substructures, the polysaccharide (P) and lipid A (L) from E. coli on the innate mechanisms of human leucocytes were examined and compared in this study. Incubation of leucocytes with LPS and L and P analogues (1 and 100 microg/ml) enhanced their biological activity in dependence on their structure. These results showed that LPS was a less active immunomodulator of leucocytes than L and P analogues isolated from E. coli strains adapted to antimicrobial agents. PMID- 18380403 TI - Sodium nitroprusside regulates the relaxation of the longitudinal muscle in the gut. AB - Nitric oxide (NO) has been shown to mediate nonadrenergic-noncholinergic relaxation in gastrointestinal (GI) smooth muscle cells. As GI smooth muscles relaxations are partly dependent on NO, we decided to investigate the effect of sodium nitroprusside (SNP) on the longitudinal muscle contraction of the isolated guinea pig ileum. Increasing concentrations of SNP (10(-10)M, 10(-9)M, 10(-8)M, 10(-7)M, 10(-6)M and 10(-5)M) reduced ileum contractions stimulated by electrical stimulation (ES) (8-76%; p < 0.05) and by acetylcholine (Ach) (23-62%; p < 0.05) significantly and in a concentration-dependent manner. Furthermore, treatment with an inhibitor of the soluble guanylate cyclase, methylene blue (10 mM), antagonized significantly the relaxing effect of SNP (0-39%; p < 0.05, p < 0.01, p < 0.001 for ES- and 4-27%; p < 0.05 for Ach-induced contractions). The results show that treatment with 1 microM manganese-containing superoxide dismutase (MnSOD) and 10 microM L-arginine (L-arg) caused a significant decrease in SNP induced relaxations (6-55%; p < 0.05, p < 0.001 and 2-46%; p < 0.05, p < 0.01 for ES- and 15-28%; p < 0.05, p < 0.01, p < 0.001 and 12-32%; p < 0.05, p < 0.01 for Ach-induced contractions, respectively). In conclusion, our data suggest that SNP, which releases NO, is able to depress longitudinal muscle contraction of the isolated guinea pig ileum, suggesting that exogenous application of NO inhibits intestinal contractions of smooth muscle cells and that cGMP mediates the response to NO. In addition, MnSOD and L-arg decreased the relaxing effect of SNP on the isolated ileum of the guinea pig. PMID- 18380405 TI - Protective efficacy of an aerosol preparation, obtained from Geranium sanguineum L., in experimental influenza infection. AB - A polyphenol-rich extract from the aerial roots of the medicinal plant Geranium sanguineum L. (PC) protected mice from mortality in the experimental influenza A/Aichi/2/68 (H3N2) virus infection. To provide evidence how a maximum therapeutic benefit can be derived of this preparation, it was inoculated by 6 different routes. It was found that the aerosol application of PC was highly effective. In the dose 5.4 mg/ml, applied according to a prophylactic-therapeutic schedule, the extract exhibited a marked protective effect. The protective index reached the value of 70.1% and the mean survival time was prolonged with 2.9-4.9 days. The lung infectious virus titres and the lung consolidation of virus infected and PC-treated animals were all reduced in comparison with control. The application of PC according to schedules, excluding the pretreatment of mice, proved that this condition was essential for protection. PMID- 18380404 TI - Rifabutin autoinduction is caused by involvement of cytochrome P450 and cholinesterase. AB - Rifabutin exhibits remarkable autoinduction of its elimination, but the mechanism behind it was not fully known. Our work showed that rifabutin administration increased the metabolism of rifabutin itself in both in vivo studies and liver perfusion, and the half-life was decreased by 71.08% and 12.74%, respectively. Further results showed that rifabutin administration increased CYP3A, CYP2D and cholinesterase levels by 87.2%, 57.3% and 65.14%, respectively. The autoinduction phenomenon of rifabutin may, therefore, be attributed to induction of cholinesterase and CYP450 isoenzymes, such as PMID- 18380406 TI - Effects of the extracts of some tropical medicinal plants on estrogen inducible yeast and Ishikawa screens, and on ovariectomized Wistar rats. AB - A total of 33 extracts from 18 Cameroonians plants were studied in two in vitro test systems to determine potential estrogenic activities. The estrogenic activities of the extracts, which have shown promising activity on both in vitro screens were further investigated in vivo on ovariectomized Wistar rats. All 33 extracts were screened in the yeast test-system. Five of these extracts, namely the ethyl acetate extract of the stem bark of Millettia conraui, the ethyl acetate extract of the stem bark of Millettia drastica, the methanol extract of the leaves of Bridelia ferruginea, the methanol extract of the roots of Pseudarthria hookeri and the methanol extract of the roots of Nauclea latifolia showed interesting estrogenic properties, and were therefore further investigated on alkaline Phosphase induction in Ishikawa cells. The extracts of Millettia conraui, Millettia drastica, Pseudarthria hoockeri and Nauclea latifolia showed significant stimulatory effects at 10 and 100 mg/ml doses. The extract of Bridelia ferruginea was not further evaluated because of its toxicity on Ishikawa cells. This stimulatory effect was completely inhibited by a combined treatment with the pure antiestrogen ICI (Faslodex, 5 x 10(-7) M). In vivo experiments showed that per os administration of 200 mg/kg bw of the extracts of Millettia conraui and Bridelia ferruginea significantly increased uterine epithelial height by 17.93% and 28.08% respectively compared with uteri of ovariectomized controls after 7 days of treatment. Uterine epithelial height of animals treated with 100 rg/kg bw/d of ethinylestradiol increased by 242.3% in the same experiment. Extracts of Nauclea latifolia and Millettia drastica had no effect on the uterine epithelial height of ovariectomised rats. 200 mg/kg bw/d of the extracts of Nauclea latifolia, Millettia drastica, Bridelia ferruginea and Millettia conraui given orally significantly increased vaginal epithelial height by 15.64%, 24.06%, 51.02% and 58.12% following the same treatment regiment compared to untreated controls. In line with these data was the finding that vaginal epithelial height and vaginal cornification in the presence of each of these extracts was more advanced than in ovariectomized controls although not as prominent as in response to ethinylestradiol treatment. These results suggest that some constituents of the extracts of Millettia conraui, Millettia drastica, Pseudarthria hookeri, Nauclea latifolia and Bridelia ferruginea may have estrogenic activity. PMID- 18380407 TI - Synthesis and biological activity of thienodiltiazem in isolated heart muscle preparations of guinea pigs. AB - The new compound thienodiltiazem was synthesized and investigated regarding structure-activity relations and calcium antagonistic properties. Isometric contraction force was measured in guinea-pig papillary muscles and chronotropic activity was studied in right atria of guinea pigs. Compared to the parent drug diltiazem (CAS 42399-41-7) the thieno derivative had a more potent negative chronotropic effect on spontaneously beating right atria and a more potent inotropic effect on papillary muscles. The negative inotropic action was reversed by increasing the extracellular calcium concentration. PMID- 18380408 TI - Interrelations between diabetes therapy, self-monitoring of blood glucose, blood glucose and non-fatal or fatal endpoints in patients with type 2 diabetes / results of a longitudinal cohort study (ROSSO 5). AB - The ROSSO study is a retrospective, longitudinal cohort study performed to obtain epidemiological data on self-monitoring of blood glucose (SMBG) in patients with type 2 diabetes and to investigate the impact of SMBG on disease-related morbidity and mortality. 3,268 patients from 192 doctor's practices in Germany were included and their data from diabetes diagnosis (between 1995 and end of 1999) till drop-out (120 died, 17 drop-outs) or end of 2003 were collected from the medical records. The mean observational time was 6.5 years; total patient years of follow-up were 21.266 years. Based on these population data, questions about the motivation of patients with type 2 diabetes to start with SMBG, the changes in diabetes therapy and blood glucose associated with SMBG and the relationship of SMBG with non-fatal or fatal events during follow-up were evaluated. Use of SMBG is significantly associated with personal and baseline conditions. Patients using SMBG are more frequently treated by an internist, more often male, have more frequently a private health insurance, exhibit less frequently arterial hypertension, are younger, have lower systolic blood pressure and higher values of fasting blood glucose (FBG), HbA1c and triglycerides at diagnosis. The start of SMBG is preceded by a steady increase in blood glucose levels. It is accompanied by an intensification of diabetes therapy and followed by a significant reduction of blood glucose in the year after start of SMBG. 67% of the patients treated with diet only before SMBG began with antidiabetic medication concomitantly with SMBG (48% with oral antidiabetic agents (OAD), 9% with insulin and 10% with OAD and insulin) and 30% of the patients treated with OAD started on additional insulin treatment in parallel with SMBG. Switching from no antidiabetic medication to OAD reduced the mean FBG levels significantly from 9.31 to 8.70 mmol/l, from no medication to insulin from 10.05 to 6.93 mmol/l and from no medication to OAD and insulin from 10.85 to 8.92 mmol/l. Similar reductions of mean FBG levels were observed for switching from OAD to insulin therapy. Patients who stayed on OAD therapy also showed a significant reduction of FBG concentrations after the start of SMBG. The hazard for non-fatal events (particularly myocardial infarction and stroke) or overall mortality was significantly reduced for patients who performed SMBG during follow-up (p<0.001). SMBG-associated changes of antidiabetic therapy may contribute to the better clinical outcome of patients with SMBG. PMID- 18380409 TI - Effect of fenofibrate on serum and tissue sialic acid levels in short-term experimental hypercholesterolemia. AB - In recent years, sialic acid is considered to be a possible marker for cardiovascular diseases. The aim of this study was to investigate the effect of two different treatment periods of fenofibrate (CAS 49562-28-9) on serum, heart and liver sialic acid levels in experimental hypercholesterolemia. Serum, heart and liver total sialic acid levels were determined by Warren's thiobarbituric acid (TBA) method and serum lipid levels by commercial kits at the end of the fenofibrate treatment for 3 and 6 weeks. Fenofibrate treatment reduced serum total sialic acid levels significantly in the control and hypercholesterolemic groups at the end of the 3rd week and only in the control group at the end of the 6th week. Serum sialic acid levels of fenofibrate-treated hypercholesterolemic rats in the 6-week period were significantly higher than those in the 3-week period. Neither the hypercholesterolemic diet nor fenofibrate had any significant effect on heart and liver sialic acid levels. In conclusion, decreased serum sialic acid levels in control and hypercholesterolemic groups by fenofibrate short-term treatment may contribute to the decreased risk of cardiovascular diseases that accompanies the hypercholesterolemic complications. The decreased serum sialic acid levels further indicate the clinical efficacy of fenofibrate as an anti-inflammatory agent. PMID- 18380410 TI - Synthesis and analgesic and anti-inflammatory activity of ethyl (6-substituted-3 (2H)-pyridazinone-2-yl)acetate derivatives. AB - A number of 6-substituted-3(2H)-pyridazinones and the corresponding methyl (6 substituted-3(2H)-pyridazinone-2-yl)acetate derivatives carrying the arylpiperazinyl structure present in potent antinociceptive agents reported in the literature were synthesized. As part of a programme a series of diverse arylpiperazine derivatives of ethyl (6-substituted-3(2H)-pyridazinone-2 yl)acetate were prepared and tested for their in vivo analgesic and anti inflammatory activity by using p-benzoquinone-induced writhing test and carrageenan-induced hind paw edema model, respectively. Side effects of the compounds were examined on gastric mucosa. None of the compounds showed gastric ulcerogenic effect compared with reference non-steroidal anti-inflammatory drugs (NSAIDs). On the basis of available data, the structure-activity relationship in the series of ethyl (6-substituted-3(2H)-pyridazinone-2-yl)acetate derivatives was also discussed. When compared to parent 6-substituted-3(2H)-pyridazinones, the new ester derivatives, for example ethyl (6-4-[(2-fluoro)phenyl]piperazine 3(2H)-pyridazinone-2-yl)acetate exhibited better analgesic and anti-inflammatory activity and a lower ulcerogenic effect. PMID- 18380411 TI - Structural basis of the anti-inflammatory activity of melatonin. AB - The anti-inflammatory activity of melatonin (CAS 73-31-4) was examined, using the rat paw edema model, and compared with the non-steroidal anti-inflammatory drug (NSAID) indometacin (CAS 53-86-1) which exerts its effects by inhibition of prostaglandin production on acute inflammation. The experiments showed that melatonin has an important effect on acute inflammatory processes acting as an inhibitor in a similar manner to indometacin. The structural interactions of melatonin with cyclooxygenase (COX), the pharmacological target of NSAIDs, were investigated using computer graphics applications. The results indicated that melatonin has an excellent steric and electronic complementarity with COX. It was found, similarly to previously studied crystal structures of protein-inhibitor complexes, that almost all interactions were of the hydrophobic type but for the typical carboxylate or electronegative group interaction, at the mouth of the active site channel, with Arg 120 and Tyr 355. Therefore, it seems possible that melatonin might bind to the active site of COX-1 and COX-2 suggesting that it may act as a natural inhibitor of the functions of cyclooxygenase modulating in a natural manner the activity of this enzyme. PMID- 18380412 TI - Glucuronidation of paracetamol by human liver microsomes in vitro / enzyme kinetic parameters and interactions with short-chain aliphatic alcohols and opiates. AB - In this study, glucuronidation of paracetamol (CAS 103-90-2) by human liver microsomes and the effects of aliphatic alcohols and opiates were investigated. Paracetamol glucuronidation was optimised for various incubation conditions. Ten different aliphatic alcohols and the opiates morphine, codeine and dihydrocodeine were analysed as inhibitors of paracetamol glucuronidation. Furthermore, the effects of paracetamol on morphine-3 and codeine glucuronidation were investigated. Enzyme kinetic analysis was carried out via determination of the parameters Km, Vmax, Ki and the type of inhibition. Except for methanol and ethanol, all Investigated alcohols inhibited glucuronidation of paracetamol. Ki values ranged between 4.59 mmol/l (n-pentanol) and 340.54 mmol/l (2-propanol). Extent of inhibition strongly depended on the structure and clearly increased with the length of the alkyl chain. All tested opiates inhibited paracetamol glucuronidation with Ki values between 4.02 mmol/l (dihydrocodeine) and 11.44 mmol/l (morphine). Paracetamol itself turned out to be an inhibitor of opiate glucuronidation. The apparent Ki values were 4.62 mmol/l (inhibition of morphine 3 glucuronidation) and 9.44 mmol/l (inhibition of codeine glucuronidation). A mixed inhibition type was determined for all substances. The in vitro studies show a great inhibition potential for the analysed substances. Transferring the results to the in vivo situation, a higher liver toxicity of paracetamol can be assumed, if concomitantly a lot of alcoholic beverages with congener alcohols- e.g. fruit schnapps or whisky--are drunk or if opiates--as analgesics or narcotics--are taken in higher doses. PMID- 18380413 TI - Synthesis of some novel quinolines and pyrimido [4,5-b] quinolines bearing A sulfonamide moiety as potential anticancer and radioprotective agents. AB - Some novel 4-(quinolin-1-yl)-benzenesulfonamide and 4-(pyrimido[4,5-b]quinolin-10 yl)-benzenesulfonamide derivatives have been synthesized. All the newly synthesized target compounds were subjected to in vitro cytotoxic screening to be evaluated for their anticancer activity against Ehrlich ascites carcinoma cells. Among these new compounds, compounds 9a, 11, 12b, 18 and, in particular, 19 showed promising in vitro cytotoxic activity compared with doxorubicin (CAS 23214 92-8) as a reference drug. Moreover, compound 8 exhibited in vivo radioprotective activity against gamma-irradiation in mice. PMID- 18380414 TI - [Pesticide residues--what does the consumer know and what should he know?]. PMID- 18380415 TI - [Insights into the German health care system]. PMID- 18380416 TI - [Is defibrillation by the laity effective?]. PMID- 18380417 TI - ["Intensive care children at home": a campaign on all fronts--the status of families with "intensive care children"]. PMID- 18380418 TI - [Handicapped children are frequently victims of violence]. PMID- 18380419 TI - [Influenza in children: what should be considered in prevention and treatment?]. PMID- 18380420 TI - [Germany needs after care: comprehensive care]. PMID- 18380421 TI - [Medical treatment error: differences between nursing personnel and physicians in readiness "error notification"]. PMID- 18380422 TI - [Relations between maturing grandchildren and their grandparents]. PMID- 18380423 TI - [Diabetes game as product of management oriented education]. PMID- 18380424 TI - Ligand binding efficiency: trends, physical basis, and implications. AB - Ligand efficiency (i.e., potency/size) has emerged as an important metric in drug discovery. In general, smaller, more efficient ligands are believed to have improved prospects for good drug properties (e.g., bioavailability). Our analysis of thousands of ligands across a variety of targets shows that ligand efficiency is dependent on ligand size with smaller ligands having greater efficiencies, on average, than larger ligands. We propose two primary causes for this size dependence: the inevitable reduction in the quality of fit between ligand and receptor as the ligand becomes larger and more complex and the reduction in accessible ligand surface area on a per atom basis as size increases. These results have far-ranging implications for analysis of high-throughput screening hits, fragment-based approaches to drug discovery, and even computational models of potency. PMID- 18380426 TI - Similarity searching and scaffold hopping in synthetically accessible combinatorial chemistry spaces. AB - Large collections of combinatorial libraries are an integral element in today's pharmaceutical industry. It is of great interest to perform similarity searches against all virtual compounds that are synthetically accessible by any such library. Here we describe the successful application of a new software tool CoLibri on 358 combinatorial libraries based on validated reaction protocols to create a single chemistry space containing over 10 (12) possible products. Similarity searching with FTrees-FS allows the systematic exploration of this space without the need to enumerate all product structures. The search result is a set of virtual hits which are synthetically accessible by one or more of the existing reaction protocols. Grouping these virtual hits by their synthetic protocols allows the rapid design and synthesis of multiple follow-up libraries. Such library ideas support hit-to-lead design efforts for tasks like follow-up from high-throughput screening hits or scaffold hopping from one hit to another attractive series. PMID- 18380427 TI - Mechanisms for the deamination reaction of cytosine with H2O/OH(-) and 2H2O/OH( ): a computational study. AB - Mechanisms for the deamination reaction of cytosine with H 2O/OH (-) and 2H 2O/OH (-) to produce uracil were investigated using ab initio calculations. Optimized geometries of reactants, transition states, intermediates, and products were determined at MP2 and B3LYP using the 6-31G(d) basis set and at B3LYP/6-31+G(d) levels of theory. Single point energies were also determined at MP2/G3MP2Large and G3MP2 levels of theory. Thermodynamic properties (Delta E, Delta H, and Delta G), activation energies, enthalpies, and free energies of activation were calculated for each reaction pathway investigated. Intrinsic reaction coordinate (IRC) analysis was performed to characterize the transition states on the potential energy surface. Seven pathways for the deamination reaction were found. All pathways produce an initial tetrahedral intermediate followed by several conformational changes. The final intermediate for all pathways dissociates to product via a 1-3 proton shift. The activation energy for the rate-determining step, the formation of the tetrahedral intermediate for pathway D, the only pathway that can lead to uracil, is 115.3 kJ mol (-1) at the G3MP2 level of theory, in excellent agreement with the experimental value (117 +/- 4 kJ mol ( 1)). PMID- 18380425 TI - Herkinorin analogues with differential beta-arrestin-2 interactions. AB - Salvinorin A is a psychoactive natural product that has been found to be a potent and selective kappa opioid receptor agonist in vitro and in vivo. The activity of salvinorin A is unusual compared to other opioids such as morphine in that it mediates potent kappa opioid receptor signaling yet leads to less receptor downregulation than observed with other kappa agonists. Our initial chemical modifications of salvinorin A have yielded one analogue, herkinorin ( 1c), with high affinity at the microOR. We recently reported that 1c does not promote the recruitment of beta-arrestin-2 to the microOR or receptor internalization. Here we describe three new derivatives of 1c ( 3c, 3f, and 3i) with similar properties and one, benzamide 7b, that promotes recruitment of beta-arrestin-2 to the microOR and receptor internalization. When the important role micro opioid receptor regulation plays in determining physiological responsiveness to opioid narcotics is considered, micro opioids derived from salvinorin A may offer a unique template for the development of functionally selective mu opioid receptor ligands with the ability to produce analgesia while limiting adverse side effects. PMID- 18380428 TI - Mixed-valent dodecanuclear vanadium cluster encapsulating chloride anions and its reaction to form a "bowl"-shaped cluster. AB - Two oxovanadium phosphonate cage compounds have been synthesized in an organic solvent, and their characterization has been done by single-crystal X-ray analysis, IR spectroscopy, and bond valence sum calculations. The simple reaction of a mixed-valent closed V12 cage system produced another quasi-closed system composed of two V6 bowl-type cages. PMID- 18380429 TI - Novel thermochromism relating to supramolecular cuprophilic interaction: design, synthesis, and luminescence of copper(I) pyrazolate trimer and polymer. AB - Solvothermal reactions of 4-(pyrid-4'-yl)-3,5-dimethylpyrazole (HPpz) with CuBr in two mixed solvents, NH3.H2O/EtOH and NH3.H2O/MeCN, afforded respectively a copper(I) trimer, [Cu(Ppz)]3(1), and a polymer, {[Cu(Ppz)]3[CuCN] 3} (2), both containing the [Cu(Ppz)]3 entity as a building block. The products were found to be photoluminescent and, more interestingly, when cooled from room temperature to 10 K, they showed a blue shift followed by a red shift (hereafter shortened to a red-after-blue shift) of emission. PMID- 18380432 TI - Activation in prochiral reaction assemblies on Pt(111). AB - Trifluoroacetophenone (TFAP) forms C O...H-C bonded dimers and trimers at room temperature on Pt(111). It is proposed that these systems mimic the prochiral carbonyl-chiral modifier interaction in the enantioselective hydrogenation of TFAP on cinchona-modified Pt catalysts. That is, the activation of TFAP in homomolecular assemblies at racemic sites is expected to be roughly the same as in the diastereomeric complex formed at chiral sites. This interpretation suggests a reason why alpha-phenyl ketones do not display a strong measured rate enhancement effect in the Orito reaction. PMID- 18380431 TI - Anisotropic assembly of Ag nanoprisms. AB - The Ag nanoprisms with controlled arrangements show distinct optical, crystallographic, and surface-enhanced Raman scattering properties depending on their orientation in the assemblies, demonstrating that the controlled assembly of anisotropic nanostructures can be utilized as a powerful tool for studying their physicochemical properties and for the creation of new classes of functional materials. PMID- 18380430 TI - Folding induced assembly of polypeptide decorated gold nanoparticles. AB - Reversible assembly of gold nanoparticles controlled by the homodimerization and folding of an immobilized de novo designed synthetic polypeptide is described. In solution at neutral pH, the polypeptide folds into a helix-loop-helix four-helix bundle in the presence of zinc ions. When immobilized on gold nanoparticles, the addition of zinc ions induces dimerization and folding between peptide monomers located on separate particles, resulting in rapid particle aggregation. The particles can be completely redispersed by removal of the zinc ions from the peptide upon addition of EDTA. Calcium ions, which do not induce folding in solution, have no effect on the stability of the peptide decorated particles. The contribution from folding on particle assembly was further determined utilizing a reference peptide with the same primary sequence but containing both D and L amino acids. Particles functionalized with the reference peptide do not aggregate, as the peptides are unable to fold. The two peptides, linked to the nanoparticle surface via a cysteine residue located in the loop region, form submonolayers on planar gold with comparable properties regarding surface density, orientation, and ability to interact with zinc ions. These results demonstrate that nanoparticle assembly can be induced, controlled, and to some extent tuned, by exploiting specific molecular interactions involved in polypeptide folding. PMID- 18380433 TI - Formation of trioctylamine from octylamine on Au(111). AB - The adsorption of octylamine on Au(111) under ultrahigh vacuum conditions is investigated. The molecules surprisingly undergo a thermally activated chemical reaction, resulting in formation of trioctylamine as confirmed both by X-ray photoelectron spectroscopy (XPS) and by comparison to the scanning tunneling microscopy (STM) signature of trioctylamine deposited directly onto the surface. PMID- 18380434 TI - Asymmetric direct aldol reaction of functionalized ketones catalyzed by amine organocatalysts based on bispidine. AB - Organocatalysts containing primary-secondary amine based on bispidine and amino acid have been designed to catalyze the asymmetric direct aldol reaction of functionalized ketones including alpha-keto phosphonates, alpha-keto esters, as well as alpha,alpha-dialkoxy ketones as aldol reaction acceptors. The corresponding products with chiral tertiary alcohols were obtained in moderate to high yields (up to 97%) and high enantioselectivities (up to 98% ee). A theoretical study of transition structures demonstrated that protonated piperidine was important for the reactivity and enantioselectivity of this reaction. PMID- 18380435 TI - Trapping of tin(II) and lead(II) homologues of carbon monoxide by a benzannulated lutidine-bridged bisstannylene. AB - The reaction of the benzannulated bisstannylene ligand 2 with Sn O or Pb O generated in situ gave the pincer complexes 3 and 4. Both complexes have been characterized by X-ray diffraction and multinuclear NMR spectroscopy. A divalent state has been found by Mossbauer spectroscopy for the tin atoms in complexes 3 and 4. PMID- 18380436 TI - Mapping the binding site of thiopeptide antibiotics by proximity-induced covalent capture. AB - Proximity-induced covalent capture (PICC) has been established for the investigation of ligand binding to composite protein/oligonucleotide target complexes. The RNA-induced attachment of the thiopeptides Thiostrepton and Nosiheptide to engineered Cys mutants of the ribosomal protein L11 was highly position selective and allowed mapping of their binding site at amino acid resolution. PMID- 18380437 TI - Nickel-catalyzed addition of C-H bonds of terminal alkynes to 1,3-dienes and styrenes. AB - The C-H bond of a terminal alkyne adds to a carbon-carbon double bond of 1,3 dienes, styrenes, and norbornene at room temperature in the presence of a nickel catalyst in regio- and stereoselective manners. Reaction of triisopropylsilylacetylene with 1-substituted 1,3-butadiene derivatives afforded hydroalkynylation products via introduction of a hydrogen atom and a triisopropylsilylethynyl group to 4- and 3-positions of the dienes, respectively. Likewise, 1-triisopropylsiloxy-1,3-butadiene, 1,3-pentadiene, 1-cyclohexen-1-yl 1,3-butadiene, and 1,3-cyclohexadiene underwent the hydroalkynylation reaction, giving the corresponding 1,4-enyne derivatives in good yields at room temperature. Reaction of p-substituted styrene with triisopropylsilylacetylene also proceeded in the presence of the nickel catalyst, giving the branched hydroalkynylation products in good yields. Norbornene gave a exo-addition product in good yield under the same reaction conditions. PMID- 18380438 TI - The mechanism of the rhodium(I)-catalyzed [2 + 2 + 1] carbocyclization reaction of dienes and CO: a computational study. AB - The rhodium(I) catalyzed [2 + 2 + 1] carbocyclization of tethered diene-enes to afford substituted hexahydropentalenones with high levels of diastereoselectivity was modeled using density functional theory. Previously, this transformation was observed to be facile, whereas the analogous bis-ene substrate could not be cyclized under any reasonable conditions. To establish a conceptual understanding of the carbocyclization mechanism and to identify the functional role of the diene fragment we analyzed the simulated reaction mechanisms using the two parent systems. We discovered a thus far unrecognized, but intuitively plausible, role of the CO ligand for controlling the electron density at the metal center, which affects the feasibility of oxidative addition and reductive elimination steps that are key components of the mechanism. Our calculations suggest that the diene moiety is unique and required because of its ability to undergo a eta(2)-->eta(4) reorganization allowing for the thermoneutral expulsion of one CO ligand, which in turn generates an electron-rich, coordinatively saturated Rh(I) center that can efficiently promote the oxidative addition with a low barrier. A number of functionalization strategies were considered explicitly to derive a rational plan for optimizing the catalysis and to expose the roles of the different components of the reactant-catalyst complex. PMID- 18380439 TI - Synthesis of homochiral dihydroxy-4-nitroisoxazolines via one-pot asymmetric dihydroxylation-reduction. AB - Herein we report a one-pot procedure to prepare a family of homochiral dihydroxy 4-nitroisoxazolines 3. This new methodology affords the title compounds in good yields, excellent enantiopurity, and without the use of chromatography. PMID- 18380440 TI - First total synthesis of (-)-achilleol B: reassignment of its relative stereochemistry. AB - The first total synthesis of (-)-achilleol B was achieved using a convergent approach with a longest linear sequence of 14 steps. Three key steps were employed, including an enantioselective Robinson annelation for the construction of the bicyclic moiety. The monocyclic synthon was prepared through a Ti(III) mediated cylization of a chiral monoepoxide obtained via asymmetric dihydroxylation of geranylacetone. The asymmetric preparation of these subunits also permitted us to achieve the enantioselective synthesis of elegansidiol, achilleol A, and farnesiferol B. PMID- 18380442 TI - Direct vinylation and difluorovinylation of arylboronic acids using vinyl- and 2,2-difluorovinyl tosylates via the Suzuki-Miyaura cross coupling. AB - General reaction conditions were developed for the Pd(0)-catalyzed Suzuki-Miyaura coupling reaction of aryl boronic acids with a simple electrophilic vinylation reagent, vinyl tosylate, providing access to styrene derivatives in good yields. The easily accessible vinyl tosylate represents a stable and less toxic alternative to the vinyl halides and the triflate/nonaflate derivatives. Furthermore, this methodology was expanded to provide a facile and straightforward approach for the introduction of a gem-difluorovinyl substituent onto an aromatic ring using the similar and also readily available 2,2 difluorovinyl tosylate as the electrophilic complement. PMID- 18380443 TI - Et2Zn-mediated rearrangement of bromohydrins. AB - A simple and highly efficient method for the rearrangement of bromohydrins mediated by Et 2Zn to synthesize carbonyl compounds was described. Various beta bromo alcohols were treated with 0.6 equiv of Et 2Zn to form a zinc complex in CH 2Cl 2 at room temperature for 2 h, followed by 1,2-migration to give the corresponding carbonyl compounds. This remarkable and clean rearrangement is general for acyclic and cyclic bromohydrins, and a variety of ring-expansive and contractive carbonyl compounds were obtained in good to excellent yields according to the feature of the starting bromohydrins. The functional group tolerance of organozinc reagents in this reaction will be useful in organic synthesis. The scope and limitations of this rearrangement process were also investigated. PMID- 18380444 TI - Testing the hypothesis that selenium deficiency is a risk factor for clozapine induced agranulocytosis in rats. AB - Clozapine is an effective atypical antipsychotic associated with a relatively high incidence of drug-induced agranulocytosis. It forms a reactive nitrenium ion metabolite upon oxidation by peripheral neutrophils and their precursors in the bone marrow. Although the mechanism of this idiosyncratic drug reaction is still unknown, the observation that it does not occur rapidly on rechallenge of patients with a history of clozapine-induced agranulocytosis suggests that it is not immune-mediated. Previous studies by other research groups had found that patients on clozapine had lower plasma and red blood cell levels of selenium. The reactive metabolite of clozapine reacts with glutathione, and therefore, it is likely that it also binds to selenocysteine-containing proteins, such as glutathione peroxidase, thioredoxin reductase, and protein disulfide isomerase. We set out to test the hypothesis that clozapine-induced agranulocytosis is associated with selenium deficiency with rats on a selenium-deficient diet. We studied the effects of clozapine on selenium levels and the effect of selenium deficiency on leukocyte and neutrophil counts and clozapine covalent binding. We did not observe any significant difference between clozapine-treated rats given a selenium-adequate or deficient diet. Therefore, it is unlikely that selenium deficiency is a major risk factor for clozapine-induced agranulocytosis. PMID- 18380445 TI - Testing the hypothesis that vitamin C deficiency is a risk factor for clozapine induced agranulocytosis using guinea pigs and ODS rats. AB - The use of clozapine is limited by a relatively high incidence of drug-induced agranulocytosis. Clozapine is oxidized by bone marrow cells to a reactive nitrenium ion. Although many idiosyncratic drug reactions are immune-mediated, the fact that patients with a history of clozapine-induced agranulocytosis do not immediately develop agranulocytosis on rechallenge suggests that some other factor may be responsible for the idiosyncratic nature of this reaction. The reactive nitrenium ion is very rapidly reduced back to clozapine by vitamin C, and many schizophrenic patients are vitamin C deficient. We set out to test the hypothesis that vitamin C deficiency is a major risk factor for clozapine-induced agranulocytosis using a vitamin C deficient guinea pig model. Although the vitamin C deficient guinea pigs did not develop agranulocytosis, the amount of clozapine covalent binding in these animals was less than we had previously observed in samples from rats and humans. Therefore, we studied ODS rats that also cannot synthesize vitamin C. Vitamin C deficient ODS rats also did not develop agranulocytosis, and furthermore, although covalent binding in the bone marrow was greater than that in the guinea pig, it was not increased in the vitamin C deficient ODS rats relative to ODS rats that had adequate vitamin C in their diet. Therefore, it is very unlikely that vitamin C deficiency is a major risk factor for clozapine-induced agranulocytosis. PMID- 18380446 TI - Fresh look at electron-transfer mechanisms via the donor/acceptor bindings in the critical encounter complex. AB - Seminal insights provided by the iconic R. S. Mulliken and his "charge-transfer" theory, H. Taube and his "outer/inner-sphere" mechanisms, R. A. Marcus and his "two-state non-adiabatic" theory, and N. S. Hush and his "intervalence" theory are each separately woven into the rich panoramic tapestry constituting chemical research into electron-transfer dynamics, and its mechanistic dominance for the past half century and more. In this Account, we illustrate how the simultaneous melding of all four key concepts allows sharp focus on the charge-transfer character of the critical encounter complex to evoke the latent facet of traditional electron-transfer mechanisms. To this end, we exploit the intervalence (electronic) transition that invariably accompanies the diffusive encounter of electron-rich organic donors (D) with electron-poor acceptors (A) as the experimental harbinger of the collision complex, which is then actually isolated and X-ray crystallographically established as loosely bound pi-stacked pairs of various aromatic and olefinic donor/acceptor dyads with uniform interplanar separations of r(DA) = 3.1 +/- 0.2 A. These X-ray structures, together with the spectral measurements of their intervalence transitions, lead to the pair of important electron-transfer parameters, H(DA) (electronic coupling element) versus lambdaT (reorganization energy), the ratio of which generally defines the odd-electron mobility within such an encounter complex in terms of the resonance stabilization of the donor/acceptor assembly [D, A] as opposed to the reorganization-energy penalty required for its interconversion to the electron-transfer state [D(+*), A(-*)]. We recognize the resonance-stabilization energy relative to the intrinsic activation barrier as the mechanistic binding factor, Q = 2H(DA)/lambdaT, to represent the quantitative measure of the highly variable continuum of inner-sphere/outer-sphere interactions that are possible within various types of precursor complexes. First, Q << 1 identifies one extreme mechanism owing to slow electron-transfer rates that result from the dominance of the intrinsic activation barrier (lambdaT) between the encounter and successor complexes. At the other extreme of Q > or = 1, the overwhelming dominance of the resonance stabilization (H(DA)) predicts the odd-electron mobility between the donor and acceptor to occur without an activation barrier such that bimolecular electron transfer is coincident with their diffusional encounter. In between lies a potentially infinite set of states, 0 < Q < 1 with opposing attractive and destabilizing forces that determine the location of the bound transition states along the reaction coordinate. Three prototypical potential-energy surfaces evolve as a result of progressively increasing the donor/acceptor bindings (H(DA)) extant in the precursor complex (at constant lambdaT). In these cases, the "outer-sphere" mechanism is limited by the weak donor/acceptor coupling that characterizes the now classical Marcus outer-sphere mechanism. Next, the "inner sphere" mechanism derives from moderate (localized) donor/acceptor bindings and includes the mechanistic concept of the bridged-activated complex introduced by Taube for a wide variety of ligand-based redox dyads. Finally, the "interior" mechanism is also another subclass of the Taube (inner-sphere) classification, and it lies at the other extreme of very fast electron-transfer rate processes (heretofore unrecognized), arising from the spontaneous annihilation of the donor/acceptor dyad to the delocalized (electron-transfer) complex as it descends barrierlessly into the chemical "black hole" that is rate-limited solely by diffusion. PMID- 18380447 TI - Impact of benchmark data set topology on the validation of virtual screening methods: exploration and quantification by spatial statistics. AB - A common finding of many reports evaluating ligand-based virtual screening methods is that validation results vary considerably with changing benchmark data sets. It is widely assumed that these data set specific effects are caused by the redundancy, self-similarity, and cluster structure inherent to those data sets. These phenomena manifest themselves in the data sets' representation in descriptor space, which is termed the data set topology. A methodology for the characterization of data set topology based on spatial statistics is introduced. The method is nonparametric and can deal with arbitrary distributions of descriptor values. With this methodology it is possible to associate differences in virtual screening performance on different data sets with differences in data set topology. Moreover, the better virtual screening performance of certain descriptors can be explained by their ability of representing the benchmark data sets by a more favorable topology. Finally it is shown, that the composition of some benchmark data sets causes topologies that lead to overoptimistic validation results even in very "simple" descriptor spaces. Spatial statistics analysis as proposed here facilitates the detection of such biased data sets and may provide a tool for the future design of unbiased benchmark data sets. PMID- 18380448 TI - Surrogate AutoShim: predocking into a universal ensemble kinase receptor for three dimensional activity prediction, very quickly, without a crystal structure. AB - "Ensemble surrogate AutoShim" is a kinase specific extension of the AutoShim docking method that solves the three traditional limitations of conventional docking: (1) it gives good correlations with affinity, (2) does not require a target protein structure, and (3) for a preprocessed company archive of 1.5 million compounds, is as fast as traditional 2D QSAR. It does require several hundred experimental IC 50 values for each new target. Original AutoShim adds pharmacophore "shims" to a crystal structure binding site. An iterative partial least squares (PLS) procedure selects the best pose, while adjusting the shim weights to reproduce IC 50 data. Surrogate AutoShim adjusts shims in one crystal structure to reproduce IC 50 data for a different kinase target. Ensemble surrogate AutoShim uses 16 structurally diverse kinase crystal structures as a "universal ensemble kinase receptor", suitable for any kinase target. The 1.5 million member Novartis screening collection has been predocked into the shimmed ensemble, so new kinase models can be built, and the entire corporate archive virtually screened, in hours rather than weeks. A kinase-biased set of 10,000 compounds, that samples the entire corporate archive, has been designed for lead discovery by iterative kinase screening. PMID- 18380449 TI - AutoShim: empirically corrected scoring functions for quantitative docking with a crystal structure and IC50 training data. AB - It has been notoriously difficult to develop general all-purpose scoring functions for high-throughput docking that correlate with measured binding affinity. As a practical alternative, AutoShim uses the program Magnet to add point-pharmacophore like "shims" to the binding site of each protein target. The pharmacophore shims are weighted by partial least-squares (PLS) regression, adjusting the all-purpose scoring function to reproduce IC 50 data, much as the shims in an NMR magnet are weighted to optimize the field for a better spectrum. This dramatically improves the affinity predictions on 25% of the compounds held out at random. An iterative procedure chooses the best pose during the process of shim parametrization. This method reproducibly converges to a consistent solution, regardless of starting pose, in just 2-4 iterations, so these robust models do not overtrain. Sets of complex multifeature shims, generated by a recursive partitioning method, give the best activity predictions, but these are difficult to interpret when designing new compounds. Sets of simpler single-point pharmacophores still predict affinity reasonably well and clearly indicate the molecular interactions producing effective binding. The pharmacophore requirements are very reproducible, irrespective of the compound sets used for parametrization, lending confidence to the predictions and interpretations. The automated procedure does require a training set of experimental compounds but otherwise adds little extra time over conventional docking. PMID- 18380450 TI - Luminescent one- and two-dimensional extended structures and a loosely associated dimer based on platinum(II)-thallium(I) backbones. AB - Neutralization reactions between (NBu4)2[ trans-Pt(C 6F5)2(CN)2] 1 and (NBu4)2[cis-Pt(C6F5)2(CN)2] 2 with TlPF 6 have been carried out, and the resulting structures of [trans,trans,trans-Tl2{Pt(C6F5)2(CN)2}.(CH3COCH3) ] n [4.(CH3COCH3)2] n and {Tl[Tl{cis-Pt(C6F5)2(CN)2}].(H2O)} n [5.(H2O)] n have been determined by X-ray crystallography. Remarkably, the change from trans to cis geometry on the platinum substrate causes a significant decrease in the Pt(II)...Tl(I) metallophilic interaction. Thus, the platinum center in the trans fragment easily connects with two Tl(I) ions forming a distorted pseudo octahedron PtTl2, which generates a final two-dimensional layered structure by secondary additional intermolecular Tl(I)...N(CN) interactions. However, the [cis Pt(C6F5)2(CN)2] (2-) fragment interacts strongly with just one Tl center leading to an extended helical [-Pt-Tl-Pt-Tl-] n(n-) chain. In this case, the second thallium center neutralizes the anionic chain mainly through Tl...N(CN) ( intra) and Tl...F(C 6F 5) (intra and inter)actions. The reaction of TlPF 6 with the monoanionic fragment (NBu4)[cis-Pt(C6F5)2(CN)(PPh2C[triple bond]CPh)] 3 yields the discrete associated dimer [Tl{cis-Pt(C6F5)2(CN)(PPh2C[triple bond]CPh)}] 2 [ 6] 2. Dimer [ 6] 2 could be described as two square pyramids with the thallium atoms in the apical positions, connected through Tl...N(cyano) interactions. The final heteropolynuclear Pt-Tl complexes, except 4 at room temperature, show bright emission in the solid state when irradiated with UV-vis radiation, in contrast to the precursors 1 and 3, which are not luminescent. This difference indicates that the emissions in 4- 6 are presumably related to the interaction between the metal centers. The Pt-Tl bonding interactions and, consequently, the emissive properties are lost in solution at room temperature, as shown by the conductivity and NMR measurements. However, variable-concentration luminescence measurements in glassy acetonitrile solutions indicate the formation of different aggregates with different degrees of Pt...Tl interactions for 4 and 5 and a dimeric structure similar to that observed in solid state for 6. PMID- 18380451 TI - Synthesis and characterization of high nuclearity iron(III) phosphonate molecular clusters. AB - Three new phosphonic acid ligands (4- (t)butylphenyl phosphonic acid, 3,5 dimethylphenyl phosphonic acid, and diphenylmethylphosphonic acid) have been synthesized and employed in search of high molecularity iron(III) clusters. The cluster compounds are characterized by single crystal X-ray diffraction and magnetic measurements. The solvothermal reaction of FeCl 3.6H 2O with diphenylacetic acid and p- (t)butylphenyl phosphonic acid resulted in an unprecedented dodecanuclear cluster [Fe 12(mu 2-O) 4(mu 3-O) 4(O 2CCHPh 2) 14(4- (t)buPhPO 3H) 6]( 1) having a double butterfly like core structure. [Fe 12(mu 2 O) 4(mu 3-O) 4(O 2CPh) 14(C 10H 17PO 3H) 6]( 2), another dodecanuclear cluster having core structure similar to 1, has been synthesized in a reaction between [Fe 3O(O 2CPh) 6(H 2O) 3]Cl and camphylphosphonic acid in the presence of triethylamine at ambient condition. 3,5-Dimethylphenyl phosphonic acid on reacting solvothermally with an oxo-centered iron triangle [Fe 3O(O 2CCMe 3) 6(H 2O) 3]Cl gives a nonanuclear cluster [Fe 9(mu 3-O) 4(O 3PPh(Me) 2) 3(O 2CCMe 3) 13]( 3) having icosahedral type core structure where three positions of the iron atoms have been replaced by phosphorus. Another nonanuclear [Fe 9(O) 3(OH) 3(O 3PCHPh 2) 6(O 2CCMe 3) 6(H 2O) 9] ( 4), having a distorted cylindrical core structure, has been synthesized in a similar solvothermal reaction between [Fe 3O(O 2CCMe 3) 6(H 2O) 3]Cl and biphenylmethyl phosphonic acid. All compounds are characterized by IR spectra, elemental analysis, as well as single crystal X-ray analysis. Magnetic measurements for all the compounds reveal that there are antiferromagnetic interactions between the metal centers. PMID- 18380452 TI - Preparation and comprehensive characterization of [Hg6(alanine)4(NO3)4].H2O. AB - A new mercury-alanine complex has been isolated from reaction mixtures of mercurous nitrate dihydrate and alanine (L and D enantiomers). The solid-state structure contains mercury(I) and mercury(II) associated by alanine ligands in a polymeric array. The disproportionation of mercury(I) to mercury(II) and mercury(0) was facilitated by alanine and is evidenced by the appearance of mercury(0) in reactions of mercury(I) with the 20 common amino acids. This complex is the first mercury(I)-amino acid complex characterized in the solid state. The compounds have been comprehensively characterized using X-ray crystallography, solid-state and solution-state nuclear magnetic resonance spectroscopy, vibrational spectroscopies, and electrospray ionization mass spectrometry. PMID- 18380453 TI - (TAML)FeIV O complex in aqueous solution: synthesis and spectroscopic and computational characterization. AB - Recently, we reported the characterization of the S = (1)/ 2 complex [Fe (V)(O)B*] (-), where B* belongs to a family of tetraamido macrocyclic ligands (TAMLs) whose iron complexes activate peroxides for environmentally useful applications. The corresponding one-electron reduced species, [Fe (IV)(O)B*] (2-) ( 2), has now been prepared in >95% yield in aqueous solution at pH > 12 by oxidation of [Fe (III)(H 2O)B*] (-) ( 1), with tert-butyl hydroperoxide. At room temperature, the monomeric species 2 is in a reversible, pH-dependent equilibrium with dimeric species [B*Fe (IV)-O-Fe (IV)B*] (2-) ( 3), with a p K a near 10. In zero field, the Mossbauer spectrum of 2 exhibits a quadrupole doublet with Delta E Q = 3.95(3) mm/s and delta = -0.19(2) mm/s, parameters consistent with a S = 1 Fe (IV) state. Studies in applied magnetic fields yielded the zero-field splitting parameter D = 24(3) cm (-1) together with the magnetic hyperfine tensor A/ g nbeta n = (-27, -27, +2) T. Fe K-edge EXAFS analysis of 2 shows a scatterer at 1.69 (2) A, a distance consistent with a Fe (IV)O bond. DFT calculations for [Fe (IV)(O)B*] (2-) reproduce the experimental data quite well. Further significant improvement was achieved by introducing hydrogen bonding of the axial oxygen with two solvent-water molecules. It is shown, using DFT, that the (57)Fe hyperfine parameters of complex 2 give evidence for strong electron donation from B* to iron. PMID- 18380454 TI - Spectral evidence and DFT calculations on the formation of Bis(2,2' bipyridine)platinum(II)-N-base adducts. AB - The formation of 1:1 adducts of Pt(bpy)2(2+) (bpy = 2,2'-bipyridine) with various N bases (B) has been ascertained in water at ambient temperature by spectrophotometric titration and electrospray ionization mass spectroscopy. A pseudo-five-coordinated complex, [Pt(bpy)2(B)](2+) with a monodentating bpy, is proposed based on density functional theory calculation. The formation constants (Kc) increase with the nucleophilicity of B except for sterically hindered N bases, indicating an associative ligand-substitution mechanism. PMID- 18380455 TI - Three-fold intramolecular ring-closing metatheses involving square-planar platinum complexes with cis-phosphorus donor ligands: syntheses, structures, and properties of parachute-like complexes. AB - Reactions of the phosphite and phosphine complexes cis-PtCl 2((PX(CH2)mCH=CH2)3)2 (X/m = O/3, O/4, O/5, -/5, -/6) with Grubbs' catalyst, followed by hydrogenations, yield cis-PtCl2(P(X(CH2)(2m+2)X)3P) (6-40%). Crystal structures establish parachute-like motifs in which one X(CH2)(2 m+2)X bridge lies roughly in the platinum coordination plane, and the others lie above and below. PMID- 18380456 TI - Synthesis, potentiometric, kinetic, and NMR Studies of 1,4,7,10 tetraazacyclododecane-1,7-bis(acetic acid)-4,10-bis(methylenephosphonic acid) (DO2A2P) and its complexes with Ca(II), Cu(II), Zn(II) and lanthanide(III) ions. AB - A cyclen-based ligand containing trans-acetate and trans-methylenephosphonate pendant groups, H 6DO2A2P, was synthesized and its protonation constants (12.6, 11.43, 5.95, 6.15, 2.88, and 2.77) were determined by pH-potentiometry and (1)H NMR spectroscopy. The first two protonations were shown to occur at the two macrocyclic ring N-CH 2-PO 3 (2-) nitrogens while the third and fourth protonations occur at the two phosphonate groups. In parallel with protonation of the two -PO 3 (2-) groups, the protons from the NH (+)-CH 2-PO 3 (2-) are transferred to the N-CH 2-COO (-) nitrogens. The stability constants of the Ca (2+), Cu (2+), and Zn (2+) (ML, MHL, MH 2L, and M 2L) complexes were determined by direct pH-potentiometry. Lanthanide(III) ions (Ln (3+)) form similar species, but the formation of complexes is slow; so, "out-of-cell" pH-potentiometry (La (3+), Eu (3+), Gd (3+), Y (3+)) and competitive spectrophotometry with Cu(II) ion (Lu (3+)) were used to determine the stability constants. By comparing the log K ML values with those of the corresponding DOTA (H 4DOTA = 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid) and DOTP (H 8DOTP = 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetramethylenephosphonic acid) complexes, the order DOTA < DO2A2P < DOTP was found for all the metal ion complexes examined here with the exception of the Ca (2+) complexes, for which the order is reversed. The relaxivity of Gd(DO2A2P) decreases between pH 2 and 7 but remains constant in the pH range of 7 < pH < 12 ( r 1 = 3.6 mM (-1) s (-1)). The linewiths of the (17)O NMR signals of water in the absence and presence of Gd(DO2A2P) (at pH = 3.45 and 8.5) between 274 and 350 K are practically the same, characteristic of a q = 0 complex. Detailed kinetic studies of the Ce (3+) and Gd (3+) complexes with DO2A2P showed that complex formation is slow and involves a high stability diprotonated intermediate Ln(H 2DO2A2P)*. Rearrangement of the diprotonated intermediate into the final complex is an OH (-) assisted process but, unlike formation of Ln(DOTA) complexes, rearrangement of Ln(H 2DO2A2P)* also takes place spontaneously likely as a result of transfer of one of the protons from a ring nitrogen to a phosphonate group. The order of the OH (-) assisted formation rates of complexes is DOTA > DO2A2P > DOTP while the order of the proton assisted dissociation rates of the Gd (3+) complexes is reversed, DOTP > DO2A2P > DOTA. (1)H and (13)C NMR spectra of Eu(DO2A2P) and Lu(DO2A2P) were assigned using two-dimensional correlation spectroscopy (2D COSY), heteronuclear multiple quantum coherence (HMQC), heteronuclear chemical shift correlation (HETCOR), and exchange spectroscopy (EXSY) NMR methods. Two sets of (1)H NMR signals were observed for Eu(DO2A2P) characteristic of the presence of two coordination isomers in solution, a twisted square antiprism (TSAP) and a square antiprism (SAP), in the ratio of ~93% and ~7%, respectively. Line shape analysis of the (1)H NMR spectra of Lu(DO2A2P) gave lower activation parameters compared to La(DOTP) for interconversion between coordination isomers. This indicates that the Ln(DO2A2P) complexes are less rigid probably due to the different size and spatial requirements of the carboxylate and phosphonate groups. PMID- 18380457 TI - Solution dynamics and gas-phase chemistry of Pd2@Sn18 4-. AB - Sn9(4-) reacts with Pd(PPh3)4 in ethylenediamine/toluene solvent mixtures in the presence of 2,2,2-cryptand to give the Pd2@Sn18(4-) cluster as the K(2,2,2, crypt)+ salt. The cluster is isostructural with Pd2@Ge18(4-) and has a nuclearity different from that of the Pt and Ni analogues, Ni2@Sn17(4-) and Pt2@Sn17(4-). The Pd2@Sn18(4-) ion has a deltahedral capsulelike structure with 40 cluster bonding electrons and is the largest free-standing polystannide characterized to date. Like Pt2@Sn17(4-), the Pd2@Sn18(4-) complex is highly dynamic in solution, showing a single (119)Sn NMR resonance indicative of an intramolecular liquidlike dynamic exchange. LDI-MS studies of the crystalline sample show extensive fragmentation and the formation of five gas-phase cluster series: Sn(x)- (1 < x < 12), PdSn(x-1) - (4 < x < 18), Pd 2Sn(x-2) - (6 < x < 21), Pd3Sn(x-3) - (8 < x < 21), and Pd 4Sn(x-4) - (13 < x < 21). The most abundant ion in the gas phase is the PdSn(10) - cluster, which presumably has an Sn(10) bicapped-square antiprismatic structure with an endohedral Pd (e.g., Ni@Pb(10)(2-)). PMID- 18380458 TI - A pH-metric, UV, NMR, and X-ray crystallographic study on arsenous acid reacting with dithioerythritol. AB - The aqueous solutions of arsenous acid with the meso and racemic forms of 1,4 dithiol-butane-2,3-diol, namely, dithioerythritol (dte) and dithiothreitol (dtt), respectively, were titrated pH-metrically in different molar ratios. The p K a values determined for As(OH) 3, and dtt were in good accordance with the literature data, and we determined for the first time the p K a value of dte. The deprotonation steps of both M (As(OH) 3 considered as a central metal ion) and H 2L components dte and dtt (considered as ligands) appeared at a higher pH in the titration curves of the ternary systems (M, H 2L, H (+)) than in the individual component. This unusual observation is explained by the condensation reactions between the reagents taking place in the pH < 8 range. In the solutions of c As(III) > 5.10 (-3) M, the precipitate formed upon mixing the arsenous acid and H 2L solutions in neutral medium, and the formation of the precipitate shifted toward acidic pH on the increase of the total concentrations. This indicated that pH-metry can follow the reactions only in an indirect way. Useful, but not satisfactory, information can be obtained by means of this method alone. Combined with NMR and UV spectroscopic measurements it is revealed that depending on the As(III)/H 2L molar ratio, different complexes form in the solutions. In the species with 1:2 composition, one of the ligands is strongly bound to the arsenic(III) probably via its two thiolate, while the second one is attached only weakly. The crystal structure of an As(III)-dte crystal of 1:1 composition, grown from ethanolic solution, shows that As(III) binds the ligand through its three p orbitals in a manner similar to that expected in aqueous solution. While the uptake of the second ligand cannot be detected by pH-metry, the decomposition of thioether bonds above pH approximately 10 is confirmed by the change in UV spectra at approximately 265 nm to be a base-consuming process. In such alkaline solutions, most probably, rearrangement of the bonding scheme occurs, resulting in ligands being bound to the arsenic(III) through the oxygen donor atoms. PMID- 18380459 TI - Spin-frustrated trinuclear Cu(II) clusters with mixing of 2(S = 1/2) and S = 3/2 states by antisymmetric exchange. 1. Dzialoshinsky-Moriya exchange contribution to zero-field splitting of the S = 3/2 state. AB - The mixing of the spin-frustrated 2(S = 1/2) and S = 3/2 states by the Dzialoshinsky-Moriya (DM) exchange is considered for the Cu 3(II) clusters with strong DM exchange coupling. In the antiferromagnetic Cu 3 clusters with strong DM interaction, the 2(S = 1/2)-S = 3/2 mixing by the in-plane DM exchange ( G x ) results in the large positive contribution 2 D DM > 0 to the axial zero-field splitting (ZFS) 2 D of the S = 3/2 state. The correlations between the ZFS 2 D DM of the excited S = 3/2 state, sign of G z and chirality of the ground-state were obtained. In the isosceles Cu 3 clusters, the in-plane DM exchange mixing results in the rhombic magnetic anisotropy of the S = 3/2 state. Large distortions result in an inequality of the pair DM parameters, that leads to an additional magnetic anisotropy of the S = 3/2 state. In the {Cu 3} nanomagnet, the in-plane DM exchange (Gx, Gy) mixing results in the 58% contribution 2 D DM to the observed ZFS 2 D of the S = 3/2 state. The DM exchange and distortions explain the experimental observation that the intensities of the electron paramagnetic resonance (EPR) transitions arising from the 2(S = 1/2) group of levels of the {Cu 3} nanomagnet are comparable to each other and are 1 order of magnitude weaker than that of the S = 3/2 state. In the ferromagnetic Cu 3 clusters, the in plane DM exchange mixing of the excited 2(S = 1/2) and the ground S = 3/2 states results in the large negative DM exchange contribution 2 D DM' < 0 to the axial ZFS 2 D of the ground S = 3/2 state. PMID- 18380460 TI - Evaluation of insecticidal activity of diterpenes and lignans from Aristolochia malmeana against Anticarsia gemmatalis. AB - The insecticidal activity of hexane extracts from the roots and leaves of Aristolochia malmeana was evaluated against Anticarsia gemmatalis larvae by topical application. Extract from the roots was the most active and caused 50% mortality in larvae at 308.4 microg/microL. From this extract, a clerodane diterpene, (-)-kolavenic acid, and three lignans, (-)-kusunokinin, (-)-hinokinin, and (8 S,8' R,9 S)-cubebin, were isolated by chromatography and partition procedures and then evaluated for their insecticidal activities either individually or in pairs. (-)-Kusunokinin showed higher activity against A. gemmatalis (LD10=9.3, LD50=230.1 microg/microL) than the crude extract, and its activity was dose-dependent, whereas the other constituents did not exhibit any significant activity. Together with (-)-kusunokinin and (-)-hinokinin, (-) copalic acid, (-)-2-oxokolavenic acid, (-)- ent-6-beta-hydroxy-copalic acid, (8 R,8' R,9 R)- and (8 R,8' R,9 S)-cubebins, (-)-fargesin, and (-)-phillygenin were isolated from the hexane extract of the leaves. The compounds were identified on the basis of spectroscopic analysis. PMID- 18380461 TI - Authentication of farmed and wild turbot (Psetta maxima) by fatty acid and isotopic analyses combined with chemometrics. AB - Fatty acid composition and stable isotope ratios of carbon (delta(13)C) and nitrogen (delta(15)N) were determined in muscle tissue of turbot (Psetta maxima). The multivariate analysis of the data was performed to evaluate their utility in discriminating wild and farmed fish. Wild (n=30) and farmed (n=30) turbot of different geographical origins (Denmark, The Netherlands, and Spain) were sampled from March 2006 to February 2007. The application of linear discriminant analysis (LDA) and soft independent modeling of class analogy (SIMCA) to analytical data demonstrated the combination of fatty acids and isotopic measurements to be a promising method to discriminate between wild and farmed fish and between wild fish of different geographical origin. In particular, IRMS (Isotope Ratio Mass Spectrometry) alone did not permit us to separate completely farmed from wild samples, resulting in some overlaps between Danish wild and Spanish farmed turbot. On the other hand, fatty acids alone differentiated between farmed and wild samples by 18:2n-6 but were not able to distinguish between the two groups of wild turbot. When applying LDA isotope ratios, 18:2n-6, 18:3n-3, and 20:4n-6 fatty acids were decisive to distinguish farmed from wild turbot of different geographical origin, while delta(15)N, 18:2n-6, and 20:1n-11 were chosen to classify wild samples from different fishing zones. In both cases, 18:2n-6 and delta(15)N were determinant for classification purposes. We would like to emphasize that IRMS produces rapid results and could be the most promising technique to distinguish wild fish of different origin. Similarly, fatty acid composition could be used to easily distinguish farmed from wild samples. PMID- 18380462 TI - Surfactant protein of the Streptomyces subtilisin inhibitor family inhibits transglutaminase activation in Streptomyces hygroscopicus. AB - Transglutaminase (TGase) is widely used in the food industry for improving protein properties by catalyzing the cross-linking of proteins. In Streptomyces, TGase is secreted as a zymogen, and an activation process has been observed in liquid culture. However, the activation mechanism remains unclear. In the present study, the TGase activation process in Streptomyces hygroscopicus was investigated by biochemical approaches. In a liquid culture, Pro-TGase was secreted and gradually was converted into active TGase during the growth period; however, in a cell-free system in which cells were removed from the liquid culture, TGase activation stalled unexpectedly. Subsequently, the TGase activation process was found to be inhibited by a TGase-activating protease inhibitor (TAPI). N-Terminal amino acid sequencing and a homology search of the purified TAPI revealed that it is a member of the Streptomyces subtilisin inhibitor (SSI) family. Furthermore, it was found that TAPI (0.1 mg/mL) decreased the surface tension of water from 72 to 60 mJ/m2 within 5 min, suggesting that it possesses surface activity. This is the first report that an SSI member functions as a surfactant protein. On the basis of these findings, a model for TAPI regulated TGase activation process was proposed. This study provides novel insights into the TGase activation process in Streptomyces. PMID- 18380463 TI - Diphenylamine metabolism in 'braeburn' apples stored under conditions conducive to the development of internal browning. AB - Diphenylamine metabolism and ethylene action were evaluated as factors influencing the development of 'Braeburn' apple internal browning and cavitation during cold storage. Apples treated with the antioxidant diphenylamine (DPA) and/or the ethylene action inhibitor 1-methylcyclopropene (1-MCP) were held at 1 degrees C for up to 6 months in air or a controlled atmosphere (CA) containing 1 kPa of O2 and 3 kPa of CO2. Cortex tissues from fruit without disorders as well as from symptomatic and asymptomatic areas of fruit with disorders were analyzed for DPA and DPA derivative content. Internal browning and cavities developed in control and 1-MCP-treated fruit stored in CA, whereas air-stored and CA fruit treated with DPA or with DPA and 1-MCP prior to storage did not develop disorders. Depending on the storage regimen and duration, less DPA was detected in 1-MCP-treated fruit. The 4-hydroxydiphenylamine (4OHDPA) content of control fruit decreased during air storage duration but increased between 2 and 4 months in CA storage. 4OHDPA content in 1-MCP-treated fruit increased with storage duration in CA but not air. N-Nitrosodiphenylamine (NODPA) was detected after 2 months in control fruit stored in air or CA and in 1-MCP-treated fruit stored in CA, and NODPA content in control fruit was higher compared to that in 1-MCP treated fruit. Accumulation of 4-methoxydiphenylamine (4MeODPA) in control fruit stored in air increased with storage duration, but 4MeODPA content did not change in 1-MCP-treated fruit stored in air or CA. 2-Nitrodiphenylamine content was reduced by prestorage treatment with 1-MCP, but storage environment and duration had no effect on its accumulation. The results indicate that CA storage increases the risk of disorder development in 'Braeburn' apples, that DPA can prevent disorder development, and that the content of DPA and DPA derivatives is influenced by storage environment and ethylene action. A clear relationship between DPA derivative formation and storage conditions that promote internal browning was not apparent. PMID- 18380464 TI - Molecularly imprinted polymer online solid-phase extraction coupled with high performance liquid chromatography-UV for the determination of three sulfonamides in pork and chicken. AB - A selective imprinted amino-functionalized silica gel sorbent was prepared by combining a surface molecular imprinting technique with a sol-gel process for online solid-phase extraction-HPLC determination of three trace sulfonamides in pork and chicken muscle. The imprinted functionalized silica gel sorbent exhibited selectivity and fast kinetics for the adsorption and desorption of sulfonamides. With a sample loading flow rate of 4 mL min (-1) for 12.5 min, enhancement factors and detection limits for three sulfonamides ( S/ N = 3) were achieved. The precision (RSD) for nine replicate online sorbent extractions of 5 microg L (-1) sulfonamides was less than 4.5%. The sorbent also offered good linearity ( r (2) > 0.99) for online solid-phase extraction of trace levels of sulfonamides. The method was applied to the determination of sulfonamides in pork and chicken muscle samples. The prepared polymer sorbent shows promise for online solid-phase extraction for HPLC determination of trace levels of sulfonamides in pork and chicken samples. PMID- 18380465 TI - Lipid-lowering and antioxidant effects of hydroxytyrosol and its triacetylated derivative recovered from olive tree leaves in cholesterol-fed rats. AB - This study was designed to test the lipid-lowering and antioxidative activities of triacetylated hydroxytyrosol compared with its native compound, hydroxytyrosol, purified from olive tree leaves. Wistar rats fed a standard laboratory diet or a cholesterol-rich diet for 16 weeks were used. The serum lipid levels, the thiobarbituric acid-reactive substances (TBARS) level, as an indicator of lipid peroxidation, and the activity of superoxide dismutase (SOD) as well as that of catalase (CAT) were examined. The cholesterol-rich diet induced hypercholesterolemia that was manifested in the elevation of total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C). Administration of hydroxytyrosol and triacetylated hydroxytyrosol (3 mg/kg of body weight) decreased the serum levels of TC, TG, and LDL-C significantly and increased the serum level of high-density lipoprotein cholesterol (HDL-C). Furthermore, the content of TBARS in liver, heart, kidney, and aorta decreased significantly when hydroxytyrosol and its triacetylated derivatives were orally administered to rats compared with those fed a cholesterol-rich diet. In addition, triacetylated hydroxytyrosol and hydroxytyrosol increased CAT and SOD activities in the liver. These results suggested that the hypolipidemic effect of triacetylated hydroxytyrosol and hydroxytyrosol might be due to their abilities to lower serum TC, TG, and LDL-C levels as well as to their antioxidant activities preventing the lipid peroxidation process. PMID- 18380466 TI - Efficient and divergent synthesis of fully substituted 1H-pyrazoles and isoxazoles from cyclopropyl oximes. AB - Efficient and divergent one-pot synthesis of fully substituted 1H-pyrazoles and isoxazoles from cyclopropyl oximes based on reaction conditions selection is reported. Under Vilsmeier conditions (POCl3/DMF), substituted 1 H-pyrazoles were synthesized from 1-carbamoyl, 1-oximyl cyclopropanes via sequential ring-opening, chlorovinylation, and intramolecular aza-cyclization. In the presence of POCl3/CH2Cl2, substituted isoxazoles were obtained from the cyclopropyl oximes via ring-opening and intramolecular nucleophilic vinylic substitution (SNV) reactions. PMID- 18380467 TI - P-glycoprotein efflux inhibition by amphiphilic diblock copolymers: relationship between copolymer concentration and substrate hydrophobicity. AB - The utilization of surfactants to increase intestinal absorption of drugs is a viable strategy that benefits from increases in drug solubilization and the potential for inhibition of P-glycoprotein (P-gp) mediated efflux. However, the effective concentration range for P-gp inhibition of most surfactants is defined over a narrow concentration range, below the critical micelle concentration (CMC), as a result of significant micelle sequestration of drug. Therefore, the objectives of these studies were to assess if association of P-gp substrates differing in hydrophobicity will impact the effective concentration range for P gp inhibition by amphiphilic diblock copolymers based on methoxypolyethylene glycol-block-polycaprolatone (MePEG-b-PCL). Comparisons between the micelle association and Caco-2 cellular accumulation were evaluated using two structurally homologous P-gp substrates, the relatively hydrophobic R-6G and the hydrophilic R-123, over concentrations above and below the CMC for MePEG-b-PCL diblock copolymers. An approximately 3.75-fold enhancement of R-123 accumulation occurred with 2 mM MePEG17-b-PCL5, compared to approximately 1.25-fold for R-6G. This decrease in the accumulation enhancement corresponds with the higher R-6G fraction (0.75) associated at 2 mM MePEG17-b-PCL5 compared with R-123 (0.25). Interestingly, R-6G accumulation was enhanced over a very broad range of MePEG17 b-PCL5 concentrations below the CMC. This was in contrast to R-123, which demonstrated no enhancement below the CMC. A similar concentration dependent accumulation profile was seen with other surfactants such as vitamin E TPGS and Cremophor EL and with two other P-gp substrates differing in hydrophobicity, the relatively hydrophobic paclitaxel and hydrophilic doxorubicin. In conclusion, the effective concentration range for surfactant mediated inhibition of P-gp appears to depend on the P-gp substrate hydrophobicity. PMID- 18380468 TI - Method of achieving desired potentiometric responses of polyacrylate-based ion selective membranes. AB - We introduce a simple procedure allowing preparation of cation-selective electrodes with poly( n-butyl acrylate)-based membranes containing different proportions of primary and interfering ions introduced already at the membrane preparation step, by using two different liphophilic salts of the same anion. With this approach the time required to achieve saturation of polyacrylate membranes with primary ions can be significantly shortened. Moreover, depending on the ratio of the primary and interfering ions introduced to the membrane cocktail, different potentiometric responses are obtained ranging from typical (with micromolar detection limit), through lower detection limits to super Nernstian ones. PMID- 18380469 TI - Differentiation and identification of recombinant human erythropoietin and darbepoetin Alfa in equine plasma by LC-MS/MS for doping control. AB - Recombinant human erythropoietin (rhEPO) and darbepoetin alfa (DPO) are protein based drugs for the treatment of anemia in humans by stimulating erythrocyte production. However, these agents are abused in human and equine sports due to their potential to enhance performance. This paper describes the first method for differentiation and identification of rhEPO and DPO in equine plasma by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The method comprised analyte extraction and enrichment by immunoaffinity separation with anti-rhEPO antibodies, dual digestion by trypsin and peptide-N-glycosidase F (PNGase F), and analysis by LC-MS/MS. Two unique deglycosylated tryptic peptides, (21)EAENITTGCAEHCSLNENITVPDTK (45) (T 5) from rhEPO and (77)GQALLVNSSQVNETLQLHVDK (97) (T 9) from DPO, were employed for differentiation and identification of rhEPO and DPO via LC retention times and major product ions. The limit of identification was 0.1 ng/mL for DPO and 0.2 ng/mL for rhEPO in equine plasma, and the limit of detection was 0.05 ng/mL for DPO and 0.1 ng/mL for rhEPO. Analyte carryover problem encountered was solved by adding 20% acetonitrile to the solvent of the sample digest to increase solubility of the peptides. This method was successfully applied to identification of DPO in plasma samples collected from a research horse following DPO administration and from racehorses out of competition in North America. Thus, it provides a powerful tool in the fight against blood doping with rhEPO and DPO in the horse racing industry. PMID- 18380470 TI - Nanoparticle-textured surfaces from spin coating. AB - Rough surfaces composed of discrete but relatively uniform nanoparticles were prepared from a lightly sulfonated polystyrene ionomer by spin coating from tetrahydrofuran (THF) or a THF/methanol mixture onto a silica surface. The particle morphology is consistent with the spinodal decomposition of the film surface occurring during spin coating. The particles are well wetted to the silica, and if heated for a long time above the ionomer's glass-transition temperature, the particles flow and coalesce into a smooth, homogeneous film. PMID- 18380471 TI - Long functionalized poly(ethylene glycol)s of defined molecular weight: synthesis and application in solid-phase synthesis of conjugates. AB - A concise synthesis of long-chain poly(ethylene glycol) (PEG) of defined molecular weight up to 29 ethyleneoxy units is described. These PEG diols were converted in a two-step synthesis into Fmoc-protected PEG amino acids, suitable as long linkers and compatible with solid-phase peptide synthesis. Long PEG chains (MW > 1000) can be readily synthesized with this method, which has the advantage of defined single molecular weight products over the comparable commercial polymers. The application of these PEG linkers to the synthesis of peptide-PEG-folate conjugates on a solid support was investigated. A method for the solid support synthesis of the targeting component of the conjugate, folic acid-cysteine, was developed, resulting in improved yields with respect to literature methods. The assembly of the peptide, PEG linker, and targeting group on solid support resulted in the synthesis of a conjugate of defined molecular weight and structure. PMID- 18380472 TI - Multivalency of Sonic hedgehog conjugated to linear polymer chains modulates protein potency. AB - A potently active multivalent form of the protein Sonic hedgehog (Shh) was produced by bioconjugation of a modified recombinant form of Shh to the linear polymers poly(acrylic acid) (pAAc) and hyaluronic acid (HyA) via a two-step reaction exploiting carboimiide and maleimide chemistry. Efficiency of the conjugation was approximately 75% even at stoichiometric ratios of 30 Shh molecules per linear HyA chain (i.e., 30:1 Shh/HyA). Bioactivity of the conjugates was tested via a cellular assay across a range of stoichiometric ratios of Shh molecules to HyA linear chains, which was varied from 0.6:1 Shh/HyA to 22:1 Shh/HyA. Results indicate that low conjugation ratios decrease Shh bioactivity and high ratios increase this activity beyond the potency of monomeric Shh, with approximately equal activity between monomeric soluble Shh and conjugated Shh at 7:1 Shh/HyA. In addition, high-ratio constructs increased angiogenesis determined by the in vivo chick chorioallantoic membrane (CAM) assay. These results are captured by a kinetic model of multiple interactions between the Shh/HyA conjugates and cell surface receptors resulting in higher cell signaling at lower bulk Shh concentrations. PMID- 18380473 TI - Predominant release of lysosomal enzymes by newborn rat microglia after LPS treatment revealed by proteomic studies. AB - Growing evidence suggest that microglia may play an important role in the pathogenesis of neurodegenerative disease including Parkinson's disease, Alzheimer's disease, and so forth. The activation of microglia may cause neuronal damage through the release of reactive oxygen species and proinflammatory cytokines. However, the early response of microglial cells remains unclear before cells can secrete the proinflammatory cytokines. Here, a time course analysis showed the earliest expression of inducible nitric oxide synthase and cyclooxygenase-2 at 3 and 24 h following lipopolysaccharide (LPS) treatment. To further define initial response proteins of microglia after LPS treatment, we utilized a novel mass spectrometry-based quantitative proteomic technique termed SILAC (for stable isotope labeling by amino acids in cell culture) to compare the protein profiles of the cell culture-conditioned media of 1 h LPS-treated microglia as compared with controls. The proteomic analysis identified 77 secreted proteins using SignalP; of these, 28 proteins were associated with lysosome of cells and 13 lysosome-related proteins displayed significant changes in the relative abundance after 1 h LPS treatment. Four proteins were further evaluated with Western blot, demonstrating good agreement with quantitative proteomic data. These results suggested that microglia first released some lysosomal enzymes which may be involved in neuronal damage process. Furthermore, ammonium chloride, which inhibits microglia lysosomal enzyme activity, could prevent microglia from causing neuronal injury. Hence, in addition to the numerous novel proteins that are potentially important in microglial activation mediated neurodegeneration revealed by the search, the study has indicated that the early release of lysosomal enzymes in microglial cells would contribute to LPS-activated inflammatory response. PMID- 18380474 TI - Determination and comparison of the Francisella tularensis subsp.novicida U112 proteome to other bacterial proteomes. AB - The proteins expressed by Francisella tularensis subsp. novicida U112 grown to midexponential phase were surveyed by nanoLC-tandem mass spectrometry (LC-MS/MS). To improve annotation of the genome and develop a technology to provide high throughput analysis of the Francisella proteome in multiple conditions, we sought to establish a fast and simple analysis that would reduce as much as possible the false discovery rate. Our survey detected expression of 63.0% of the predicted proteome from the stable condition of growth in rich medium available at (www.francisella.org). On the basis of detection of essential proteins, we estimated coverage to be approximately 80% of the actual expressed proteome. This suggests that no less than 70% of the proteins could be expressed in this condition. This analysis revealed two previously unidentified protein coding open reading frames and validated 50% of the proteins annotated as hypothetical. On the basis of results of the screen to detect essential proteins, not all proteins expressed provide a measurable contribution to F.t. novicida growth in this condition. Comparison of this protein profile with other profiles previously published suggested that the genome size and number of genes involved in regulation have little effect on the number of proteins expressed in a given stable condition. PMID- 18380475 TI - The reductive desulfurization of Met and Cys residues in bovine RNase A is associated with trans lipids formation in a mimetic model of biological membranes. AB - Damage to bovine pancreatic RNase A, due to the H* atom and/or solvated electron attack at protein sulfur-containing residues, was investigated by Raman spectroscopy and mass spectrometry techniques. To the already known desulfurization process affecting Met residues, novel reactivity was observed involving disulfide moieties, leading to the chemical transformation of Cys into Ala residues. Mapping experiments demonstrated that desulfurization selectively affected Met79, Cys110, Cys58 and Cys72 during first stages of reaction. While this reaction was performed on protein species added to large unilamellar vescicles, desulfurization yielded sulfur radicals able to induce a cis-trans isomerization of lipids at the onset of irradiation. These findings reveal new scenarios on reactions generated by radical stressing conditions, suggesting the need for specific assays and for future investigations to detect these modifications in proteins and lipids within challenged cells. PMID- 18380476 TI - Proteomics of two cultivated mushrooms Sparassis crispa and Hericium erinaceum provides insight into their numerous functional protein components and diversity. AB - Mushroom can be defined as a macrofungus with a distinctive fruiting body. Mushrooms of class Basidiomycete are primarily wood degradation fungi, but serve as food and a part of traditional medicine used by humans. Although their life cycle is fairly well-established, the information on the molecular components, especially proteins are very limited. Here, we report proteomics analysis of two edible mushrooms (fruiting bodies) Sparassis crispa and Hericium erinaceum using one- and two-dimensional gel electrophoresis (1-DGE and 2-DGE) based complementary proteomics approaches. 1-DGE coupled with liquid chromatography and mass spectrometry identified 77 (60 nonredundant proteins) and 121 (88 nonredundant proteins) proteins from S. crispa and H. erinaceum, respectively. 2 DGE analysis revealed 480 and 570 protein spots stained with colloidal coomassie brilliant blue in S. crispa and H. erinaceum, respectively. Of the 71 and 115 selected protein spots from S. crispa and H. erinaceum 2D gel blots on polyvinyldifluoride (PVDF) membranes, respectively, 29 and 35 nonredundant proteins were identified by N-terminal amino acid sequencing. Identified nonredundant proteins from 1- or 2-DGE belonged to 19 functional categories. Twenty-one proteins were found common in both S. crispa and H. erinaceum proteomes, including 14-3-3 protein and septin. Together this study provides evidence for the presence of a large number of functionally diverse proteins, expressed in the fruiting body of two economically important mushrooms, S. crispa and H. erinaceum. Data obtained from 1-DGE and 2-DGE analyses is accessible through the mushroom proteomics portal http://foodfunc.agr.ibaraki.ac.jp/mushprot.html. PMID- 18380477 TI - Involvement of p21 and FasL in induction of cell cycle arrest and apoptosis by neochamaejasmin A in human prostate LNCaP cancer cells. AB - Neochamaejasmin A ( 1), a biflavonoid isolated from the roots of a traditional Chinese medicine, Stellera chamaejasme L., was shown to inhibit cellular (3)H thymidine incorporation (IC 50 12.5 microg/mL) and subsequent proliferation of human prostate cancer LNCaP cells. Treatment of LNCaP cells with low doses of 1 (< or =6.25 microg/mL) suppressed DNA-binding activities of the transcription factors NFkappaB and AP-1 to the promoter of cyclin D and also inhibited expression of the cell cycle regulatory proteins cyclin D, proliferating cell nuclear antigen, and nucleolin, thus arresting cells in G 1 phase of the cell cycle. A lengthy exposure with higher doses of 1 (> or =12.5 microg/mL) revealed the production of reactive oxygen species, dissipation of the mitochondrial membrane potential, up-regulation of cyclin-dependent kinase inhibitor p21, and induction of cell apoptosis. An aggregation of Fas-procaspase 8-procaspase 3 and p21-procaspase 3 proteins by coimmunoprecipitation, immunoblotting analysis, and MALDI-mass spectrometry indicated the involvement of Fas and p21 in 1-mediated cytotoxicity, and pretreatment of cells with antisense FasL oligonucleotides partially abolished apoptosis. Thus, 1 blocked cell cycle progression at the G 1 phase by activating the p21 protein and ultimately promoting the Fas-caspase 8 caspase 3 apoptotic machinery. PMID- 18380479 TI - Design and effective synthesis of the first 4-aza-2,3-didehydropodophyllotoxin rigid aminologue: a N-methyl-4-[(3,4,5-trimethoxyphenyl)amino)]-1,2 dihydroquinoline-lactone. AB - The first N1-alkyl-4-amino-1,2-dihydroquinoline-lactone has been prepared by a five-step sequence in a 51% overall yield via the corresponding furo[3,4 b]quinolin-1(3H)-one. A new practical synthesis of this intermediate was carried out using versatile, commercially available starting materials and constitutes the shortest and highest yielding route. These synthetic pathways could be widened with a view toward the preparation of different substituted derivatives, which could be considered as rigid aminologues of 4-aza-2,3 didehydropodophyllotoxins. PMID- 18380478 TI - Aurocitrin and related polyketide metabolites from the wood-decay fungus Hypocrea sp. BCC 14122. AB - The known gentisaldehyde antibiotic aurocitrin (1), its cis-olefinic isomer (2), a salicylaldehyde analogue (3), two new benzofuran derivatives (4 and 5), and a new dihydroisocoumarin (6) were isolated from the wood-decay fungus Hypocrea sp. BCC 14122. The structures were elucidated primarily by NMR and mass spectroscopic analyses. PMID- 18380480 TI - Palladium-catalyzed intramolecular C-arylation of benzylic carbon: synthesis of 3 benzoxazolylisoindolinones by a sequence of Ugi-4CR/postfunctionalization. AB - Submitting Ugi-adduct 4 to two consecutive metal-catalyzed intramolecular reactions, namely copper-catalyzed O-arylation and palladium-catalyzed C arylation of benzylic carbon developed in the course of this study, affords the benzoxazolylisoindolinones in good to excellent yields. PMID- 18380481 TI - Fine-tuning of radiolysis induced emission by variable substitution of donor /acceptor-substituted tetrakis(arylethynyl)benzenes. AB - Emission from charge recombination between radical cations and anions of various tetrakis(arylethynyl)benzenes (TAEBs) was measured during pulse radiolysis in benzene (Bz). The formation of TAEB in the excited singlet state ((1)TAEB*) can be attributed to the charge recombination between TAEB (*+) and TAEB (*-), which is initially generated from the radiolytic reaction. It was found that the charge recombination between TAEB (*+) and TAEB (*-) gave (1)TAEB* as the emissive species but not excimers because of the large repulsion between substituents caused by the rotation around C-C single bonds. Since donor-/acceptor-substituted TAEBs possess three types of charge-transfer pathways (linear-conjugated, cross conjugated, and "bent"-conjugated pathways between the donor and acceptor substituents through the ethynyl linkage), the emission spectra of (1)TAEBs* with intramolecular charge transfer (ICT) character depend on the substitution pattern and the various types of donor and acceptor groups during pulse radiolysis. Through control of the substitution pattern (e.g., the position of the nitrogen atom within the pyridine ring or the number of acceptors per arene ring of the regioisomeric donor-/acceptor-substituted TAEBs with donating N, N-dibutylamino and accepting pyridine unit (N1-9) and those with donating N, N-dibutylamino and accepting one (F1-3), two trifluoromethyl (F4-6), or perfluorinated arene (F7-9) units), fine-tuning of radiolysis induced emission color can be achieved. PMID- 18380482 TI - Novel gold(III) polymer-supported catalyst for indole library synthesis. PMID- 18380483 TI - Synthesis and biological evaluation of novel sulfonanilide compounds as antiproliferative agents for breast cancer. AB - Combinatorial chemistry approaches facilitate drug discovery processes and result in structural modifications of lead compounds that enhance pharmacological activity, improve pharmacokinetic properties, or reduce unwanted side effects. Epidemiological and animal model studies have suggested that nonsteroidal anti inflammatory drugs (NSAIDs) can act as chemopreventive agents. The cyclooxygenase 2 (COX-2) inhibitor nimesulide shows anticancer effects in several cancer cell lines via COX-2-dependent and -independent mechanisms. The molecular structure of nimesulide was used as a starting scaffold to design novel sulfonanilide analogs and examine the structural features that contribute to this anticancer effect. A systematic combinatorial chemical approach was used to generate diversely substituted sulfonanilide derivatives that were tested for their effects on the proliferation of human breast cancer cells. Structure-function analysis indicated that the inhibition of cell growth by compounds derived from the novel sulfonanilides required a bulky terminal phenyl ring, a methanesulfonamide, and a hydrophobic carboxamide moiety. PMID- 18380484 TI - Analysis of the vapor-liquid-solid mechanism for nanowire growth and a model for this mechanism. AB - The vapor-liquid-solid (VLS) mechanism is most widely employed to grow nanowires (NWs). The mechanism uses foreign element catalytic agent (FECA) to mediate the growth. Because of this, it is believed to be very stable with the FECA-mediated droplets not consumed even when reaction conditions change. Recent experiments however differ, which suggest that even under cleanest growth conditions, VLS mechanism may not produce long, thin, uniform, single-crystal nanowires of high purity. The present investigation has addressed various issues involving fundamentals of VLS growth. While addressing these issues, it has taken into consideration the influence of the electrical, hydrodynamic, thermodynamic, and surface tension effects on NW growth. It has found that parameters such as mesoscopic effects on nanoparticle seeds, charge distribution in FECA-induced droplets, electronegativity of the droplet with respect to those of reactive nanowire vapor species, growth temperature, and chamber pressure play important role in the VLS growth. On the basis of an in-depth analysis of various issues, a simple, novel, malleable (SNM) model has been presented for the VLS mechanism. The model appears to explain the formation and observed characteristics of a wide variety of nanowires, including elemental and compound semiconductor nanowires. Also it provides an understanding of the influence of the dynamic behavior of the droplets on the NW growth. This study finds that increase in diameter with time of the droplet of tapered nanowires results primarily from gradual incorporation of oversupplied nanowire species into the FECA-mediated droplet, which is supported by experiments. It finds also that optimum compositions of the droplet constituents are crucial for VLS nanowire growth. An approximate model presented to exemplify the parametric dependency of VLS growth provides good description of NW growth rate as a function of temperature. PMID- 18380485 TI - Extracting superconducting single-crystal Nb mesowires out of NbSe2 by a crystal lattice collapse method. AB - The author reports a conceptually new approach to superconducting niobium (Nb) mesowires in high yields from layered compounds (NbSe2) by solid-state pyrolysis. High-resolution transmission electron microscopy and selected-area electron diffraction demonstrate that the mesowires are single crystalline, grow along the [001] direction, and have specific facets. Unlike the previous electrodeposition routes within templates, a possible vapor-solid interaction mechanism was first proposed for component-selective epitaxial growth of one-dimensional Nb lattices where vapor-assisted collapse of NbSe2 crystal structure occurs through disposing of Se vapor phase matrix at enhanced temperatures. PMID- 18380486 TI - Expansion of human neural precursor cells in large-scale bioreactors for the treatment of neurodegenerative disorders. AB - The transplantation of in vitro expanded human neural precursor cells (hNPCs) represents a potential new treatment alternative for individuals suffering from incurable neurodegenerative disorders such as Parkinson's disease (PD) and Huntington's disease (HD). However, in order for cell restorative therapy to have widespread therapeutic significance, it will be necessary to generate unlimited quantities of clinical grade hNPCs in a standardized method. We report here that we have developed a serum-free medium and scale-up protocols that allow for the generation of clinical quantities of human telencephalon-derived hNPCs in 500-mL computer-controlled suspension bioreactors. The average hNPC aggregate diameter in the bioreactors was maintained below a target value of 500 microm by controlling the liquid shear field. The human cells, which were inoculated at 10(5) cells/mL, exhibited a doubling time of 84 h, underwent a 36-fold expansion over the course of 18 days, and maintained an average viability of over 90%. The bioreactor-derived hNPCs retained their nestin expression following expansion and were able to differentiate into glial and neuronal phenotypes under defined conditions. It has also been demonstrated that these hNPCs differentiated to a GABAergic phenotype that has recently been shown to be able to restore functional behavior in rat models of HD and neuropathic pain (Mukhida, K. et al. Stem Cells 2007; DOI 10.1634/stemcells.2007-0326). This study demonstrates that clinical quantities of hNPCs can be successfully and reproducibly generated under standardized conditions in computer-controlled suspension bioreactors. PMID- 18380487 TI - Generation of local concentration gradients by gas-liquid contacting. AB - We present a generic concept to create local concentration gradients, based on the absorption of gases or vapors in a liquid. A multilayer microfluidic device with crossing gas and liquid channels is fabricated by micromilling and used to generate multiple gas-liquid contacting regions, separated by a hydrophobic membrane. Each crossing can acts as both a microdosing and microstripping region. Furthermore, the liquid and gas flow rate can be controlled independently of each other. The focus of this conceptual article is on the generation of pH gradients, by locally supplying acidic or basic gases/vapors, such as carbon dioxide, hydrochloric acid, and ammonia, visualized by pH-sensitive dyes. Stationary and moving gradients are presented in devices with 500-microm channel width, depths of 200-400 microm, and lengths of multiple centimeters. It is shown that the method allows for multiple consecutive switching gradients in a single microchannel. Absorption measurements in a microcontactor with the model system CO2/water are presented to indicate the dependence of gas absorption rate on channel depth and residence time. Achievable concentration ranges are ultimately limited by the solubility of used components. The reported devices are easy to fabricate, and their application is not limited to pH gradients. Two proof of principles are demonstrated to indicate new opportunities: (i) local crystallization of NaCl using HCl vapor and (ii) consecutive reactions of ammonia with copper(II) ions in solution. PMID- 18380488 TI - Assay to screen for molecules that associate with Alzheimer's related beta amyloid fibrils. AB - Small molecules that bind to aggregated forms of Abeta peptides show promise as potential in vivo labeling agents for the diagnosis and monitoring of Alzheimer's disease. A major challenge in developing potential imaging agents that target Abeta is to rapidly identify and evaluate the association of molecules with insoluble deposits of aggregated Abeta peptides. This paper describes a simple, parallel method to rapidly screen libraries of molecules for their ability to associate with fibrils formed from synthetic Abeta peptides by monitoring their ability to inhibit the interaction of a monoclonal anti-Abeta IgG with these fibrils. We demonstrate that this assay can detect the association of small molecules with Abeta fibrils at concentrations of small molecule in the nanomolar to millimolar range. By comparing results from the screening of a small set of 30 compounds, we illustrated that this assay can rapidly analyze the relative affinity of small molecules for Abeta fibrils and identified eight compounds that can bind to Abeta fibrils at <20 microM concentrations. Significant advantages of this assay are (1) the ability to screen structurally diverse molecules without requiring them to have specific spectroscopic or radiolabeled properties, (2) the ability to estimate the percentage of the surface of the fibrils covered by the small molecules, and (3) the ability to detect the association of small molecules that potentially bind to different sites along the fibril axis. This assay also has minimal requirements for equipment or specialized facilities and should, therefore, be useful for both academic and industrial laboratories. PMID- 18380490 TI - Density functional study of adenine tetrads with N6-H6...N3 hydrogen bonds. AB - The structure and energy of A-tetrads with N6-H6...N3 H-bonds was studied using B3LYP and BH&H density functional theory. The planar A-tetrad with C(4h) symmetry is more stable than the nonplanar structures at C4 and S4 symmetry. This structure corresponds to a local energy minimum. The energies of the structures with N6-H6...N1 and N6-H6...N7 H-bonds studied previously are of similar magnitude. Structures of A-tetrad complexes with sodium and potassium were most stable at S4 symmetry, and similarly, sandwich complexes consisting of two tetrads and a single cation were most stable at S8 symmetry. Relative energies of sandwich complexes with different symmetries obtained with the B3LYP and BH&H methods were quite different. BH&H overestimates the interaction energies between hydrogen-bonded neighbor bases relative to B3LYP. PMID- 18380489 TI - Self-sealed vertical polymeric nanoporous-junctions for high-throughput nanofluidic applications. AB - We developed a reliable but simple integration method of polymeric nanostructure in a poly(dimethylsiloxane) (PDMS)-based microfluidic channel, for nanofluidic applications. The Nafion polymer junction was creased by infiltrating polymer solution between the gaps created by mechanical cutting, without any photolithography or etching processes. The PDMS can seal itself with the heterogeneous polymeric nanoporous material between the PDMS/PDMS gap due to its flexibility without any (covalent) bonding between PDMS and the polymer materials. Thus, one can easily integrate the nanoporous-junction into a PDMS microchip in a leak-free manner with excellent repeatability. We demonstrated nanofluidic preconcentration of proteins (beta-phycoerythrin) using the device. Because the polymeric junction spans across the entire microchannel height, the preconcentration was achieved with high-pressure field or even in large channels, with the dimensions of 1000 microm width x 100 microm depth. PMID- 18380491 TI - Stress dependence of paramagnetic point defects in amorphous silicon oxide. AB - Room-temperature red cathodoluminescence (CL) emission (R band) arising from the paramagnetic point-defect population present in amorphous silicon oxide (SiOx) has been characterized with respect to its shift upon applied stress, according to a piezo-spectroscopic (PS) approach. The R band (found at around 630 nm) originates from nonbridging oxygen hole centers (NBOHC; Si-O*) generated in the presence of oxygen-excess sites. It is shown that reliable stress assessments can be obtained in silica glass with a relatively high spatial resolution, provided that appropriate spectroscopic procedures are developed to precisely extract from the CL spectrum the shift upon stress of the R band, isolated from other partly overlapping bands. Macroscopic and microscopic PS calibration procedures are shown to lead to consistent results on silica materials with different chemical characteristics and, thus, with different intrinsic defect populations. In addition, quantitative calibrations of both electron probe size and luminescence emission distribution within the electron probe are given. As an application of the PS technique, the magnitude of the residual stress piled up (mainly due to a thermal expansion mismatch) at a sharp silica/silicon interface has been characterized by taking into account the gradient in defect population developed as a function of distance from the interface. In the Results and Discussion section, brief comments are offered regarding the possible impact of highly spatially resolved stress assessments in silica glass upon the development of new materials and advanced electronic devices. PMID- 18380492 TI - HN2(2A') electronic manifold. II. Ab initio based double-sheeted DMBE potential energy surface via a global diabatization angle. AB - A double-sheeted double many-body expansion potential energy surface is reported for the coupled 12A'/22A' states of HN2 by fitting about 6000 ab initio energies. All crossing seams are described to their full extent on the basis of converged results. The lowest adiabatic sheet is fitted with a rmsd of 0.8 kcal mol-1 with respect to the calculated energies up to 100 kcal mol-1 above the absolute minimum, and the topology of the first excited-state investigated with the aid of the upper adiabatic sheet. A new scheme that overcomes obstacles in previous diabatization methods for modeling global double-sheeted potential energy surfaces is also reported. The novel approach uses a global diabatization angle which allows the diabats to mimic both the crossing seams and atom-diatom dissociation limits. PMID- 18380493 TI - Two-photon excitation fluorescence correlation spectroscopy of diffusion for Gaussian-Lorentzian volumes. AB - Fluorescence correlation spectroscopy (FCS) is valuable in many scientific domains where diffusion plays a fundamental role. One important experimental realization is based on fluorescence induced by two-photon excitation (TPE). In comparison with one-photon excitation (OPE), TPE-FCS defines better the interrogation volume and the background noise is sensibly reduced. Within this context and for overfilled objective lenses, the three-dimensional Gaussian (3DG) approximation, according to which the spectroscopic interaction is spatially defined by Gaussian profiles only, guarantees a simple analytical data interpretation. By contrast, the volume illuminated by the laser beam focused with partially filled objective lenses follows a Gaussian-Lorentzian (GL) distribution that is taken into account by means of numerical methods only. Here we show that contrary to common belief, the assumption of a GL volume does not hamper analytical treatment of TPE-FCS. Differences and similarities in comparison with the 3DG approximation are discussed. PMID- 18380494 TI - Effective two-photon absorption cross section of heteroaromatic quadrupolar dyes: dependence on measurement technique and laser pulse characteristics. AB - The linear and nonlinear optical properties of the heteroaromatic push-pull-push two-photon absorbing dye N-methyl-2,5-bis[1-(N-methylpyrid-4-yl)ethen-2-yl] pyrrole ditriflate (PEPEP) are reported. The determination of the two-photon absorption (TPA) cross-section spectrum has been performed with different techniques: femtosecond TPA-white light continuum probe experiments, two-photon induced fluorescence, and open aperture Z-scan measurements using both nanosecond and femtosecond laser pulses. The measured TPA cross sections and their wavelength dispersion show a marked dependence on the parameters of the laser pulses and on the measurement technique employed. These properties are discussed in terms of the different microscopic mechanisms that can contribute to the multiphoton absorption processes, with different weight depending on the measurement conditions and on the photophysical parameters of the dye. PMID- 18380496 TI - Theoretical study on the electronic states of formylcarbene (HC.CHO) and triplet ketene (H2CCO) and the ultraviolet absorption spectra attributable to these molecules. AB - The adiabatic energies, vibrational frequencies, and geometries of the ground and excited electronic states of formylcarbene and the triplet electronic states of ketene are calculated employing the state-of-the art ab initio methods. With the help of these calculations, certain ultraviolet (UV) absorption bands observed in the flash photolysis of oxazole and iso-oxazole are assigned to formylcarbene and triplet ketene. PMID- 18380495 TI - Quantum chemical and kinetic study of formation of 2-chlorophenoxy radical from 2 chlorophenol: unimolecular decomposition and bimolecular reactions with H, OH, Cl, and O2. AB - This study investigates the kinetic parameters of the formation of the chlorophenoxy radical from the 2-chlorophenol molecule, a key precursor to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCCD/F), in unimolecular and bimolecular reactions in the gas phase. The study develops the reaction potential energy surface for the unimolecular decomposition of 2-chlorophenol. The migration of the phenolic hydrogen to the ortho-C bearing the hydrogen atom produces 2-chlorocyclohexa-2,4-dienone through an activation barrier of 73.6 kcal/mol (0 K). This route holds more importance than the direct fission of Cl or the phenolic H. Reaction rate constants for the bimolecular reactions, 2 chlorophenol + X --> X-H + 2-chlorophenoxy (X = H, OH, Cl, O2) are calculated and compared with the available experimental kinetics for the analogous reactions of X with phenol. OH reaction with 2-chlorophenol produces 2-chlorophenoxy by direct abstraction rather than through addition and subsequent water elimination. The results of the present study will find applications in the construction of detailed kinetic models describing the formation of PCDD/F in the gas phase. PMID- 18380497 TI - Evaluation of the enthalpy of formation, proton affinity, and gas-phase basicity of gamma-butyrolactone and 2-pyrrolidinone by isodesmic reactions. AB - The knowledge of thermochemical parameters such as the enthalpy of formation, gas phase basicity, and proton affinity may be the key to understanding molecular reactivity. The obtention of these thermochemical parameters by theoretical chemical models may be advantageous when experimental measurements are difficult to accomplish. The development of ab initio composite models represents a major advance in the obtention of these thermochemical parameters, but these methods do not always lead to accurate values. Aiming at achieving a comparison between the ab initio models and the hybrid models based on the density functional theory (DFT), we have studied gamma-butyrolactone and 2-pyrrolidinone with a goal of obtaining high-quality thermochemical parameters using the composite chemical models G2, G2MP2, MP2, G3, CBS-Q, CBS-4, and CBS-QB3; the DFT methods B3LYP, B3P86, PW91PW91, mPW1PW, and B98; and the basis sets 6-31G(d), 6-31+G(d), 6 31G(d,p), 6-31+G(d,p), 6-31++G(d,p), 6-311G(d), 6-311+G(d), 6-311G(d,p), 6 311+G(d,p), 6-311++G(d,p), aug-cc-pVDZ, and aug-cc-pVTZ. Values obtained for the enthalpies of formation, proton affinity, and gas-phase basicity of the two target molecules were compared to the experimental data reported in the literature. The best results were achieved with the use of DFT models, and the B3LYP method led to the most accurate data. PMID- 18380498 TI - Confocal Raman studies of Mg(NO3)2 aerosol particles deposited on a quartz substrate: supersaturated structures and complicated phase transitions. AB - Individual Mg(NO3)2 aerosol particles deposited on a quartz substrate were investigated by confocal Raman spectroscopy. With decreasing the relative humidity (RH) from 92.0% to 1.8%, Raman spectra were obtained of Mg(NO3)2 droplets with water-to-solute molar ratios (WSRs) from 43.1 to 5.2, as well as of amorphous particles. At WSR < 6.0, contact ion pairs between Mg2+ and NO3(-) occurred abundantly, while at RHs of 2.2% and 1.8% with even lower WSRs, amorphous particles appeared with quasi-lattice structures. Two components, one at 3259.0 cm(-1) (C1) and the other at approximately 3480.0 cm(-1) (C2), were resolved for the water O-H stretching envelope through nonlinear curve fittings. The area ratio of C1 to C2, that is, A1/A2, declined with the decrease of WSR, reflecting the breakage of strong hydrogen bonds induced by the hydration of NO3( ). Curve fittings were also carried out for the water O-H stretching envelope of NaNO3 droplets. The value of A1/A2 for Mg(NO3)2 droplets was always higher than that for NaNO3 droplets at the same WSR, indicating a much stronger "structure making" effect of Mg2+ than of Na+. In the efflorescence process, aerosol particles followed different paths of phase transition from droplets to Mg(NO3)2.6H2O or amorphous states. Reversing somewhat the phase transitions in the efflorescence process, aerosol particles dissolved into droplets with the increase of RH in the deliquescence process. Heterogeneous particles prepared by dehydrating Mg(NO3)2.6H2O were investigated by the depth profiling technique. About 15 h later, the main body of particles changed into Mg(NO3)2.2H2O, a small quantity of Mg(NO3)2.6H2O scattered around particle edges, and some particles were in amorphous states. About 10 days later, a new solid phase occurred on particle surfaces, while the interiors were still Mg(NO3)2.2H2O. With increasing the RH to approximately 11%, significant Mg(NO3)2.6H2O formed on particle surfaces, covering the interior Mg(NO3)2.2H2O. PMID- 18380499 TI - Hydration-dependent structural deformation of guanine in the electronic singlet excited state. AB - Theoretical study was performed to investigate how the degree of hydration affects the structures and properties of the canonical form (keto-N9H) of guanine in the ground and lowest singlet pipi* excited state. This work is the continuation of our earlier work where we have studied the hydration of guanine in the first solvation shell with one, three, five, and six water molecules. In the present investigation, we have considered 7-13 water molecules in hydrating guanine. Ground-state geometries were optimized at the Hartree-Fock level, whereas the configuration interaction-singles (CIS) method was used for the excited-state geometry optimization. The 6-311G(d,p) basis set was used in all calculations. The harmonic vibrational frequency analysis was used to determine the nature of the optimized ground- and excited-state potential energy surfaces; all geometries were found to be minima at the respective potential surfaces. It was found that the degree of hydration has a significant influence on the excited state structural nonplanarity of guanine. It is expected that excited-state dynamics of guanine will depend on the degree of hydration. Ground- and excited state geometries of selected hydrated species were also optimized in the bulk water solution using the polarizable continuum model (PCM). It was found that bulk water solution generally does not have significant influence on the structure of the hydrated species. Effects of hydration on different stretching vibrations in the ground and excited states are also discussed. PMID- 18380500 TI - New indole-based chromophore-containing main-chain polyurethanes: architectural modification of isolation group, enhanced nonlinear optical property, and improved optical transparency. AB - Three indole-based nonlinear optical (NLO) chromophores with changeable isolation groups were successfully introduced into the polymer backbone to yield a series of main-chain polyurethanes. Thanks to the main-chain structure and the advantages of the indole-based chromophores, all of the polymers show excellent transparency, good processability, thermal stability, and relatively good NLO effects. The obtained experimental results indicated that the polymer backbone, in addition to the linked isolation moieties, could act as isolation spacers in some special cases. The tested NLO results demonstrated that the isolation groups with apparent site-isolation effect might not benefit the macroscopic NLO effect of the resultant polymers. PMID- 18380501 TI - Microhydration of the guanine-cytosine (GC) base pair in the neutral and anionic radical states: a density functional study. AB - A density functional study of the effects of microhydration on the guanine cytosine (GC) base pair and its anion radical is presented. Geometries of the GC base pair in the presence of 6 and 11 water molecules were fully optimized in the neutral (GC-nH2O) and anion radical [(GC-nH2O)*-] (n = 6 and 11) states using the B3LYP method and the 6-31+G** basis set. Further, vibrational frequency analysis at the same level of theory (B3LYP/6-31+G**) was also performed to ensure the existence of local minima in these hydrated structures. It was found that water molecules surrounding the GC base pair have significant effects on the geometry of the GC base pair and promote nonplanarity in the GC base pair. The calculated structures were found to be in good agreement with those observed experimentally and obtained in molecular dynamics (MD) simulation studies. The water molecules in neutral GC-nH2O complexes lie near the ring plane of the GC base pair where they undergo hydrogen bonding with both GC and each other. However, in the GC anion radical complexes (GC-nH2O, n = 6, 11), the water molecules are displaced substantially from the GC ring plane. For GC-11H2O*-, a water molecule is hydrogen-bonded with the C6 atom of the cytosine base. We found that the hydration shell initially destabilizes the GC base pair toward electron capture as a transient anion. Energetically unstable diffuse states in the hydration shell are suggested to provide an intermediate state for the excess electron before molecular reorganization of the water molecules and the base pair results in a stable anion formation. The singly occupied molecular orbital (SOMO) in the anion radical complexes clearly shows that an excess electron localizes into a pi orbital of cytosine. The zero-point-energy (ZPE-) corrected adiabatic electron affinities (AEAs) of the GC-6H2O and GC-11H2O complexes, at the B3LYP/6-31+G** level of theory, were found to be 0.74 and 0.95 eV, respectively. However, the incorporation of bulk water as a solvent using the polarized continuum model (PCM) increases the EAs of these complexes to 1.77 eV. PMID- 18380502 TI - Synthesis and photophysical properties of conjugated polymers with pendant 9,10 anthraquinone units. AB - We have synthesized and investigated the photophysical properties of a series of electron-donor conjugated copolymers with pendant electron-acceptor units. The copolymers consist of diethynyl-1,4-phenylene, fluorene, or phenylene rings alternating with a phenylene unit bearing a pendant 9,10-anthraquinone moiety. The pendant donor-acceptor polymers were designed to have different optical pi pi* band gaps, while the oxidation potential of the polymer backbone remains approximately constant in the series. The reduction potential of the donor acceptor polymers is associated with the pendant acceptor units. This leads to the special situation that the electrochemical gap between oxidation and reduction potentials is constant, while the optical band gap decreases, going from PPP, via PPF, to PPE. This design is used to study the effect of the optical gap on the photoinduced electron-transfer reaction that occurs between the main chain electron donor and the pendant acceptor, while the same polymer architecture and energy of the charge separated state are maintained. Fluorescence and photoinduced absorption spectroscopy are used to study the electron transfer following photoexcitation in relation to solvent polarity and in thin solid films. For the fluorene-phenylene alternating copolymer, intramolecular photoinduced electron transfer occurs in the Marcus optimal region. PMID- 18380503 TI - Investigations of coupling characters in ionic liquids formed between the 1-ethyl 3-methylimidazolium cation and the glycine anion. AB - In this study, novel ionic liquids formed between the 1-ethyl-3-methylimidazolium cation [emim]+ and the glycine anion [Gly]- have been investigated theoretically. The relevant geometrical characteristics, energy properties, the characters of the intermolecular hydrogen bonds (H bonds), and the possibility of proton transfer as well as IR characteristics have been systematically discussed. The natural bond orbital (NBO) and atoms in molecule (AIM) analyses have also been applied to understand the nature of the interactions between ionic pairs in ionic liquids. The most stable geometries have been determined by analyzing the relative energies and interaction energies, where the C-H...O intermolecular H bonds involving the protons attached to the imidazolium ring have been found to possess partial covalent character in nature. Electron transfers from the lone pairs of the carbonyl O atom of [Gly]- to the C-H antibonding orbital of the [emim]+ can explain the elongation and red shift of the C-H stretching frequency. The interaction modes are more favorable when the carbonyl O atoms of [Gly]- interact with the C2-H of the imidazolium ring and the C-H of the methyl group through the formation of double H bonds. The origin of the high stability of the amino acid ionic liquids observed experimentally may be attributed to the nonexistence of the proton-transferred products (neutral pairs) together with the large energy needed for separation of the ionic pairs. Additionally, the characteristics of the IR spectra have been analyzed to demonstrate the variants of the molecular structure of the [emim]+[Gly]- ionic liquids. PMID- 18380504 TI - Pressure-stiffened Raman phonons in group III nitrides: a local bond average approach. AB - It has long been puzzling regarding the atomistic origin of the pressure-induced Raman optical phonon stiffening that generally follows a polynomial expression with coefficients needing physical indication. Here, we show that an extension of the bond-order-length-strength correlation mechanism and a local bond average approach to the pressure domain have led to an analytical solution to connect the pressure-induced Raman optical phonon stiffening directly to the bonding identities of the specimen and the response of the bonding identities to the applied stimulus. It is found that the pressure-induced blue-shift of Raman optical phonons arises from the bond compression and energy storage exerted by the compressive stress. Agreement between predictions and experimental measurements lead to the clarification of the detailed form for the polynomial coefficients, which provide an atomistic understanding of the physical mechanism of the external pressure induced energy gain, thermally induced bond expansion, as well as means of determining the mode atomic cohesive energy in a specimen. PMID- 18380505 TI - Molecular modeling and simulation of conjugated polymer oligomers: ground and excited state chain dynamics of PPV in the gas phase. AB - The ground and excited state dynamics of poly(p-phenylenevinylene) (PPV) chains is studied through an implementation of mixed quantum/classical molecular dynamics simulation. The model used in the simulations combines the semiempirical Pariser-Parr-Pople (PPP) Hamiltonian to treat the pi molecular electronic structure with a mechanical force field capturing all other aspects. Nuclear degrees of freedom are treated classically. We first validate the model by simulating PPV chains of various length, and evaluate the absorption spectra. The thermal disorder contribution to the breadth of the first absorption band is estimated to be 0.2 eV at T = 300 K. To investigate the relationship between the emission and chain conformation, we simulate an isolated ten unit chain of PPV in the ground and the lowest excited state. The emission spectrum, red-shifted with respect to absorption of about 0.2 eV as found in experiments, shows a structured line shape that we relate to the photoinduced CC bond distortions. In accord with earlier studies, the exciton self-traps in the middle of the chain. We introduce two collective variables that reflect geometrical distortion, and find these to be effective in describing the contribution of chain conformation to the emission spectrum. The collective variables are also shown to be effective in describing the bond relaxation dynamics after photoexcitation. Such a relaxation is found to occur in approximately 100 fs and is guided by a compensatory release of energy between the double and single bonds in the vinylene junctions and p-phenyl rings. Finally, we find that the chain has a very slight preference for a more planar conformation in the excited state, compared to the ground state. However, the thermal motions induce the chain to explore out-of-plane conformations in both the ground and the excited states with an amplitude significantly greater than this difference. PMID- 18380506 TI - Molecular force field for ionic liquids IV: trialkylimidazolium and alkoxycarbonyl-imidazolium cations; alkylsulfonate and alkylsulfate anions. AB - This is the fourth article of a series that describes the parametrization of a force field for the molecular simulation of common ionic liquids within the framework of statistical mechanics. The force field was developed in the spirit of the OPLS-AA model and is thus oriented toward the calculation of equilibrium thermodynamic and structural properties in the condensed (liquid) phase. The ions modeled in the present paper are cations of the 1,2,3-trialkylimidazolium and alkoxycarbonyl imidazolium families and alkylsulfate and alkylsulfonate anions. As in previous publications, the force field is built in a stepwise manner that allows, for example, the construction of models for an entire family of cations or anions, with alkyl side chains of different length. Because of the transferability of the present force field, the ions studied here can be combined with those reported in our three previous publications to create a large variety of ionic liquids that can be studied by molecular simulation. The extension of the force field was validated by comparison of simulation results with the corresponding crystal structure and liquid density experimental data. PMID- 18380507 TI - The effective concentration of unbound ink anchors at the molecular printboard. AB - Self-assembled monolayers terminating in beta-cyclodextrin cavities can be used to bind ink molecules and so provide a molecular printboard for nanopatterning applications. Multivalent or multisite binding strengthens the attachment of large inks and provides more robust patterns. In the present work we use computer simulations to probe the behavior of functionalized dendrimer inks at the printboard. We performed a series of long 10 ns fully atomistic molecular dynamics (MD) simulations to measure the effective local concentration of unbound ink anchor groups at the printboard for a variety of binding modes and also for the partial unbinding prerequisite for ink diffusion on the printboard. These simulations allow us to describe the conformational space occupied by partially bound inks and estimate the likelihood of an additional binding interaction. Furthermore, by simulating the shift from a divalent to monovalent binding mode we show that the released anchor quickly moves to the periphery of the dendrimer binding hemisphere but then reapproaches the printboard and remains in the vicinity of alternative binding sites. Secondary electrostatic interactions between the protonated dendrimer core and hydroxyl groups at the entrance to the beta-cyclodextrin cavities give "flattened" dendrimer binding orientations and may aid dendrimer diffusion on the printboard, allowing the dendrimer to "walk" along the printboard by switching between different partially bound states and minimizing complete unbinding to bulk solution, crucial for the application of the printboard in, for example, medical diagnostics. PMID- 18380508 TI - Alkyl chain length dependence of the self-organized structure of alkyl substituted phthalocyanines. AB - The alkyl chain length on alkyl-substituted phthalocyanines (C(n)OPc) dependence of their self-organized structures was examined in this study. STM results indicated that the symmetry of ordered structures decreased as the alkyl chain became longer, with the exception of C(6)OPc, which preferentially formed a quasi 3-fold symmetrical structure. This could be explained by the fact that the C(n)OPc molecules are most likely to form densely packed structures. With C(n)OPc, when n = 4 to 10, the self-organized structures were dependent on the competition between how densely the molecules were arranged and how loose the intermolecular interaction energy was, caused by the formation of the densely packed structure. However, with C(n)OPc, when n = 10-18, the molecules tended to form densely packed structures by reducing the symmetry, even though the C(n)OPc molecules were distorted. When C(12)OPc and cobalt phthalocyanine were coadsorbed, the mixed system exhibited a four-fold symmetrical structure, which is rarely observed in C(12)OPc. PMID- 18380509 TI - Chemical force titration of plasma polymer-modified PDMS substrates by using plasma polymer-modified AFM tips. AB - Plasma polymerization has gained increasing attention in surface functionalization. We use here chemical force titration to characterize PDMS (polydimethylsiloxane) substrates modified by maleic anhydride-pulsed plasma polymerization. The coating is hydrolyzed to promote the formation of dicarboxylic acid groups. To enhance the variation of the adhesion forces as a function of pH, we use AFM tips modified in the same way as the substrates. The pH-dependent adhesion measurements are performed at different KCl concentrations. The dicarboxylic nature of the maleic acid groups clearly emerges from the force titration curves. The surface pK(a) values (pK(a1) = 3.5 +/- 0.5 and pK(a2) = 9.5 +/- 0.5) of the dicarboxylic acids are evaluated from low electrolyte concentration solutions. The values are shifted toward higher pK(a) values when compared to maleic acid in solution. The first pK(a) appears in the titration force curve for low salt concentration as a peak. This peak changes to a sigmoidal shape at higher salt concentrations. The appearance of a peak is attributed to the formation of strong hydrogen bonds between the tip and the substrate as reported in the literature. The effect of the ionic strength on the force curves is explained by the condensation of counterions on the carboxylate groups. At high pH, the adhesion force almost vanishes. On the approach, at high pH, one first observes repulsion between the tip and the substrate, which varies exponentially with the tip/substrate distance. The decay length of this repulsion force is in good agreement with theoretical predictions of the Debye length, attesting to the electrostatic nature of the interactions. We also find that the replacement of monovalent cation K(+) by the divalent cation Ca(2+) leads to significant changes in the force titration curve at high pH where the dicarboxylic groups are fully ionized. We observe that the adhesion force no longer vanishes at high pH but even slightly increases with pH, an effect that is explained by Ca(2+) ions bridging between two carboxylate groups. PMID- 18380510 TI - Porphyrin-based photocatalytic lithography. AB - Photocatalytic lithography couples light with photoreactive coated mask materials to pattern surface chemistry. We excite porphyrins to create radical species that photocatalytically oxidize, and thereby pattern, chemistries in the local vicinity. The technique advantageously is suited for use with a wide variety of substrates. It is fast and robust, and the wavelength of light does not limit the resolution of patterned features. We have patterned proteins and cells to demonstrate the utility of photocatalytic lithography in life science applications. PMID- 18380511 TI - Experimental studies on the adsorption of two surfactants on solid-aqueous interfaces: adsorption isotherms and kinetics. AB - A quartz crystal microbalance with dissipation (QCM-D) was used to measure the adsorption from aqueous solutions of CTAB (cationic) and C(12)E(6) (nonionic) surfactants on gold and silica surfaces. QCM-D allows for the determination of adsorption isotherms and also the monitoring of the dynamics of adsorption in real time. By considering the atomic-scale roughness of the solid surfaces and the surface area per head group at the air/water interface, our experiments indicate that at bulk concentrations above the critical micelle concentration adsorbed C(12)E(6) forms a monolayer-like structure on both surfaces and CTAB yields a bilayer-like structure. Although our measurements do not allow us to discriminate between the morphology of the aggregates (i.e., between flat monolayers, hemicylinders, or hemispheres in the case of C(12)E(6) and between flat bilayers, cylinders, or spheres in the case of CTAB), these results are particularly significant when compared to recent QCM-D data reported by Macakova et al. (Macakova, L.; Blomberg, E.; Claesson, P. M. Langmuir 2007, 23, 12436). These authors reported that QCM-D overestimates the amount of CTAB adsorbed on silica by as much as 30-40% as a result of entrapped water. Our analysis suggests that the effect of entrapped solvent is not as important as previously assumed and, in fact, QCM-D may not overestimate the amount of CTAB adsorbed when roughness is considered. Results for the kinetics of adsorption suggest that the aggregate structure as well as whether micelles are present may influence the adsorption mechanism. We discuss our results in the perspective of molecular theories for both the equilibrium and kinetics of surfactant adsorption. PMID- 18380512 TI - Macrophase and microphase separations for surfactants adsorbed on solid surfaces: a gauge cell monte carlo study in the lattice model. AB - By combining the gauge cell method and lattice model, we study the surface phase transition and adsorption behaviors of surfactants on a solid surface. Two different cases are considered in this work: macrophase transition and adsorption in a single-phase region. For the case of macrophase transition, where two phases coexist, we investigate the shape and size of the critical nuclei and determine the height of the nucleation barrier. It is found that the nucleation depends on the bulk surfactant concentration. Our simulations show that there exist a critical temperature and critical adsorption energy, below which the transition from low-affinity adsorption to the bilayer structure shows the characteristic of a typical first-order phase transition. Such a surface phase transition in the adsorption isotherm is featured by a hysteresis loop. The hysteresis loop becomes narrower at higher temperature and weaker adsorption energy and finally disappears at the critical value. For the case where no macrophase transition occurs, we study the adsorption isotherm and microphase separation in a single phase region. The simulation results indicate that the adsorption isotherm in adsorption processes is divided into four regions in a log-log plot, being in agreement with experimental observations. In this work, the four regions are called the low-affinity adsorption region, the hemimicelle region, the morphological transition region, and the plateau region. Simulation results reveal that in the second region the adsorbed monomers aggregate and nucleate hemimicelles, while adsorption in the third region is accompanied by morphological transitions. PMID- 18380513 TI - Nanotexturing of polystyrene surface in fluorocarbon plasmas: from sticky to slippery superhydrophobicity. AB - In this work plasma etching processes have been studied to roughen and fluorinate polystyrene surface as an easy method to achieve a superhydrophobic slippery character. Radiofrequency discharges have been fed with CF(4)/O(2) mixtures and the effect of the O(2):CF(4) ratio, the input power, and the treatment duration have been investigated in terms of wettability, with focus on sliding performances. For this purpose, surface morphological variations, evaluated by means of scanning electron microscopy and atomic force microscopy, together with the chemical assessment by X-ray photoelectron spectroscopy, have been correlated with water contact angle hysteresis and volume resolved sliding angle measurements. Results indicate that by increasing the height and decreasing the density of the structures formed by etching, within a tailored range, a transition from sticky to slippery superhydrophobicity occurs. A short treatment time (5 min) is sufficient to obtain such an effect, provided that a high power input is utilized. Optimized surfaces show a unaltered transparency to visible light according to the low roughness produced. PMID- 18380514 TI - Direct time-resolved detection of singlet oxygen in zeolite-based photocatalysts. AB - Singlet oxygen has been characterized spectroscopically as a product of the exposure of suspensions of zeolites containing oxidation catalysts. Spectroscopic and lifetime studies show that a part of the singlet oxygen formed reacts within the zeolite porous structure, while a significant fraction escapes and becomes available for reaction in the bulk media. The liquid phase plays a key role in determining intra- and extracavity dynamics. PMID- 18380516 TI - Snail bioaccumulation of triclocarban, triclosan, and methyltriclosan in a North Texas, USA, stream affected by wastewater treatment plant runoff. AB - Grazing by freshwater snails promotes nutrient turnover in algal communities. Grazed algal compartments may include antimicrobial agents and metabolites, such as triclocarban (TCC), triclosan (TCS), and methyltriclosan (MTCS), which are incompletely removed by wastewater treatment plant (WWTP) processing. The present study quantifies snail bioaccumulation factors (BAFs) for TCC, TCS, and MTCS at the outfall of Pecan Creek (TX, USA), the receiving stream for the city of Denton (TX, USA) WWTP. Helisoma trivolvis (Say) is ubiquitous and thrives under standard laboratory conditions, leading to its choice for this bioaccumulation study in conjunction with Cladophora spp. Along with providing substrate for epiphytic growth, Cladophora spp. provide a source of food and shelter for H. trivolvis. After being caged for two weeks, algae and snails were collected from the WWTP outfall, along with water-column samples, and analyzed by isotope dilution gas chromatography-mass spectrometry for TCS and MTCS and by liquid chromatography mass spectrometry for TCC. Algal and snail samples were analyzed before exposure and found to be below practical quantitation limits for all antimicrobial agents. Triclocarban, TCS, and MTCS in water samples were at low-ppt concentrations (40 200 ng/L). Triclocarban, TCS, and MTCS were elevated to low-ppb concentrations (50-300 ng/g fresh wt) in caged snail samples and elevated to low-ppb concentrations (50-400 ng/g fresh wt) in caged algal samples. Resulting snail and algal BAFs were approximately three orders of magnitude, which supports rapid bioaccumulation among algae and adult caged snails at this receiving stream outfall. The results further support TCC, TCS, and MTCS as good candidate marker compounds for evaluation of environmental distribution of trace WWTP contaminants. PMID- 18380520 TI - Acute, sublethal exposure to a pyrethroid insecticide alters behavior, growth, and predation risk in larvae of the fathead minnow (Pimephales promelas). AB - The present study determined the effects of environmentally relevant, short-term (4-h) exposure to the pyrethroid insecticide esfenvalerate on mortality, food consumption, growth, swimming ability, and predation risk in larvae of the fathead minnow (Pimephales promelas). Acute effect concentrations were determined, and in subsequent experiments, fish were exposed to the following measured sublethal concentrations: 0.072, 0.455, and 1.142 microg/L of esfenvalerate. To measure growth rates (% dry wt/d), 8-d-old fathead minnows were exposed to esfenvalerate for 4 h, then transferred to control water and held for 7 d. Food consumption and abnormal swimming behavior were recorded daily. Additional behavioral experiments were conducted to evaluate how esfenvalerate affects the optomotor response of the fish. To quantify predation risk, esfenvalerate-exposed fathead minnow larvae were transferred to 9.5-L aquaria, each containing one juvenile threespine stickleback (Gasterosteus aculeatus). Sticklebacks were allowed to feed for 45 min, after which the number of surviving minnows was recorded. No mortality occurred during 4-h exposures to esfenvalerate, even at nominal concentrations of greater than 20 microg/L. Delayed mortality (50%) was observed at 2 microg/L after an additional 20 h in clean water. Fish exposed to 0.455 and 1.142 microg/L of esfenvalerate exhibited impaired swimming and feeding ability as well as reduced growth compared to fish exposed to 0.072 microg/L and controls. Predation risk also was significantly increased for larvae exposed to 0.455 and 1.142 microg/L of esfenvalerate. These results demonstrate that larval fish experiencing acute exposures to sublethal concentrations of this insecticide exhibit significant behavioral impairment, leading to reduced growth and increased susceptibility to predation, with potentially severe consequences for their ecological fitness. PMID- 18380521 TI - Predicting survival of grass shrimp (Palaemonetes pugio) exposed to naphthalene, fluorene, and dibenzothiophene. AB - The composition and persistence of dissolved polycyclic aromatic hydrocarbons (PAHs) released to the water column during oil spills are altered by weathering, tidal transport, and addition of dispersants. Conventional toxicity effect metrics, such as the median lethal concentration (LC50), are inaccurate predictors of mortality from all toxicant exposure duration/concentration combinations likely to occur during spills. In contrast, survival models can predict the proportions of animals dying as a consequence of exposures differing in duration and intensity. Extending previous work with ethylnaphthalene, dimethylnaphthalene, and phenanthrene, survival time models were developed that include exposure duration and concentration to predict time to death for grass shrimp (Palaemonetes pugio). Two additional PAHs (naphthalene and fluorene) and a heterocyclic aromatic hydrocarbon (dibenzothiophene) were evaluated for the present study. Preliminary explorations of these models confirmed that quantitative structure- activity regression models were possible for predicting survival model parameters from compound characteristics. Conventional 48-h LC50s also were calculated for the compounds and combined with published LC50s to predict relative PAH toxicity to P. pugio based on octanol-water partitioning. PMID- 18380522 TI - Development of a brain nucleus involved in song production in zebra finches (Taeniopygia guttata) is disrupted by Aroclor 1248. AB - We studied whether polychlorinated biphenyls (PCBs) may alter the development of song control brain nuclei in zebra finch (Taeniopygia guttata) offspring of pulse exposed hens. We orally administered 40 microg of Aroclor 1248 to adult female finches before egg laying. When the progeny were 50 d old, we measured the volumes of the song control nuclei robustus arcopallialis (RA) and higher vocal center (HVC) using light microscopy. Both male and female progeny of exposed birds had a significantly smaller RA than control birds (36 and 16%, respectively; p < or = 0.05). The HVC did not differ in either sex between exposed and control groups. Perhaps impaired development of RA was caused by PCB action on steroid receptors. We conclude that animals living in contaminated areas may be at risk of neurological damage in hormone-sensitive brain areas and that changes in brain nuclei related to song may be a sensitive indicator of low level PCB exposure. PMID- 18380523 TI - Basin-wide impacts of compounds with estrogen-like activity on longnose dace (Rhinichthys cataractae) in two prairie rivers of Alberta, Canada. AB - Environmental compounds with estrogen- or antiestrogen-like activity can enter rivers from multiple sources, including municipal wastewater and agricultural runoff. We used longnose dace (Rhinichthys cataractae) to investigate exposure to compounds with estrogen-like activity, which we measured in water at multiple sites in the Oldman and Bow rivers (AB, Canada). We evaluated changes in vitellogenin mRNA with quantitative reverse transcription-polymerase chain reaction, then compared vitellogenin levels to sex ratios and fish performance indices to assess how exposure to compounds with estrogen-like activity affects longnose dace populations. Vitellogenin levels were elevated at least 59 to 110 km downstream of municipalities. In the Oldman River, increased vitellogenin expression and female-biased sex ratios suggest severe endocrine disruption, likely resulting from the combined impacts of municipal wastewater, agriculture, and large cattle operations within the basin. In the Bow River, municipal wastewater may be the major source of compounds with estrogen-like activity that affect longnose dace. The sex ratios were not heavily skewed, as in the more agriculturally influenced Oldman River. We detected organic contaminants in river samples at every site, but the highest concentrations were found downstream of municipalities and in areas with intense agriculture. Vitellogenin levels and sex ratios of longnose dace suggest basin-wide exposure to compounds with estrogen like activity. Our results demonstrate that it is important to assess rivers at large spatial scales to detect fully the impacts of municipal wastewater and agriculture on fish populations. PMID- 18380524 TI - Satisfaction with dental appearance in 8-9 years-old children. Validation of COAS questionnaire for Italian-speaking children and evaluation of social and geographical context. AB - AIM: This study was designed to adapt the original English-language COAS (Children's Orthodontic Attitude Survey) for third- grade schoolchildren to the Italian cultural environment and to investigate its properties in typical populations, as well as to evaluate children self-perception of their dental appearance. STUDY DESIGN: The COAS questionnaire for third-grade schoolchildren was translated and culturally adapted for Italian-speaking children. The Italian version of the questionnaire was tested on 169 (73 females, 96 males) children. Test-retest reliability was assessed on 34 children one week after the first administration. We also analysed correlations between social status and questionnaire findings. METHODS: All children filled in the questionnaire and then they were clinically examined by three residents. The clinical parameters were correlated with the questionnaire findings to evaluate children's satisfaction with their dental appearance. RESULTS: Ninety-five per cent of children thought it was important to have straight teeth and 87 per cent considered that crooked teeth were ugly. Comparison with clinical parameters showed a statistically significant correlation between crowding and overjet and some answers. Urban children have a better opinion on braces: they would like to have braces and they think they need braces statistically more than rural subjects. CONCLUSION: The Italian version of the modified-COAS questionnaire had a very good reliability. Social status and geographical context play a very important role in children's satisfaction with dental appearance. Children with different social context demonstrate they have very different approaches towards their dental aspect and braces. PMID- 18380525 TI - Laser in paediatric dentistry: patient acceptance of hard and soft tissue therapy. AB - AIM: This study aimed to evaluate the laser therapy efficacy in paediatric dentistry, considering the subjective tolerance and acceptance of therapy in children needing both dental and soft tissue treatments. METHODS: A group of 50 patients from 6 to 12 years of age, needing both hard and soft tissue therapy was selected and treated, without anesthesia, with an Er,Cr:YSGG laser 2780 nm and an Er:YAG laser 2940 nm for a total of 100 treatments performed; before and after the treatment the patients experience was tested with Wong-Baker modified facial image scale. RESULTS: The study showed a good percentage of acceptance and tolerance of both laser treatments: a success rate of 90 % for hard tissues and 63% for soft tissues was obtained; for the total 100 treatments the acceptance was of 75%. CONCLUSION: The Erbium lasers are very effective in paediatric dentistry and are good treatment options. PMID- 18380526 TI - Preschool children and relative analgesia: satisfaction grading through a verbal questionnaire. AB - AIM: The aim of the study was to assess 100 preschool children's satisfaction grading of relative analgesia (RA) after completing dental treatment by collecting their opinion on this technique through a verbal questionnaire. METHODS: After completing dental care a simple verbal questionnaire (3 questions) was administered by the operator. The questionnaire investigated: 1) patient's satisfaction about the sedation treatment; 2) patient's agreement to re experiment the technique and 3) patient's emotions while sedated. Moreover, following treatment, each child was invited to make a drawing on the experience. RESULTS: Data obtained were classified in 3 groups: group 1 (87% of children) appreciated RA and would agree to repeat the experience; group 2 (4% of the sample) did not answer the verbal questionnaire and group 3 (9%) did not enjoy the sedation technique. Only 15 children completed a drawing; conducting a psychological analysis through C.R. Rogers' theory of Person Centred Approach coupled with the handwriting analysis methodology defined by Girolamo Moretti, positive features were found in the majority of the drawings (13 out of 15). CONCLUSION: The majority (87%) of the sample appreciated to experience nitrous oxide inhalation sedation and would undergo a further appointment under RA. PMID- 18380527 TI - Correlation between dental maturation and bone growth markers in paediatric patients. AB - AIM: Many studies have been made to evaluate the growth, development and maturation of cervical vertebrae. These studies make clear how those vertebrae have growth patterns and centre of ossification similar to those in carpal bones, therefore cervical vertebrae can be used to determine bone age. The aim of the present study is the observation of cervical vertebrae in cephalometric radiographs to determine the bone age in a paediatric population and relate it to dental maturation. MATERIALS AND METHODS: The sample was composed of 203 children from Madrid, aged between 2 and 10 years. There were 86 males and 117 females, in each case a panoramic was taken to determine dental calcification using Demirjian method and a cephalometric radiograph was taken to evaluate cervical vertebrae maturation. Different vertical and horizontal dimensions were measured to determine maturation. RESULTS: The results show that there is a positive relationship between age and size of cervical vertebrae. The average horizontal measurements of the third, fourth and fifth cervical vertebrae presented statistically significant differences in size (p<0.05), being larger in boys than in girls. Statistical correlation was found between dental maturation and increased left vertical measures from third (C3VI) and fourth cervical vertebrae (C4VI), as well as middle vertical measure from fourth cervical vertebra (C4VMD). CONCLUSION: Significant changes in size were observed only in vertical measurements and the predictive measurements for dental maturation were C4VI, C4VMD and C3VI. PMID- 18380528 TI - Evaluation of customer satisfaction at the Department of Paediatric Dentistry of "Sapienza" University of Rome. AB - AIM: Healthcare facility management requires the optimisation of the quality of services offered. METHODS: The Authors adopted a questionnaire as a means to assess customer satisfaction and needs. The Questionnaire has been designed to address adults, i.e., parents or guardians of children attending the Paediatric unit as patients. To estimate their degree of satisfaction, either regarding the environment where children are treated or about the direct interactions between the parents and the structure (waiting room, waiting time, treatment time and time needed to make the payments, costs, etc.), the questionnaire was submitted to a sample of approximately 600 customers, between March and June of 2005. RESULTS: On one hand results provide a pleasant confirmation on customers' perception of the service, especially regarding the direct relationship between the parties; on the other, they highlight aspects that could be improved (waiting room, optimisation of waiting time), distinguishing between factors that need extra funding to be implemented, and those that, instead, could be simply improved through a better and more efficient organisation of labour and time. CONCLUSION: The present analysis and previous experiences confirm that appraisal of the degree of customers' satisfaction by means of questionnaires is a valid and necessary instrument for the quality improvement of a healthcare service. Doing so, economic and human resources can be more efficiently allocated. PMID- 18380529 TI - Osteopathic manipulative treatment (OMT) effects on mandibular kinetics: kinesiographic study. AB - AIM: The aim of this study was to evaluate the effects of Osteopathic Manipulative Treatment (OMT) on mandibular kinematics in TMD patients. METHODS: The study was conduced on 28 children with non-specific TMD symptoms, limited mouth opening, history of trauma (delivery trauma, accident trauma). Patients were randomly divided into two groups: an OMT group (study group) and a no intervention group (control group). All subjects underwent a first kinesiographic recording to evaluate the amplitude and velocity of maximal opening-closing movements. Study group patients underwent a second kinesiographic recording 2 months after OMT. Control group patients were submitted to a control kinesiographic recording six months after the first one. Kinesiographic tracings were acquired using the K7I system. RESULTS/STATISTICS: The kinesiographic data of the study group showed a moderate statistically significant difference (p<.07) of maximal mouth opening (MO) parameter and a high statistically significant difference (p<.03) of maximal mouth opening velocity (MOV) parameter. No statistically significative difference (null hypothesis confirmed) of kinesiographic parameters in the control group was observed. CONCLUSION: The results of this study suggest that OMT can induce changes in the stomatognathic dynamics, offering a valid support in the clinical approach to TMD. Multifactorial genesis of chronic disorders is also confirmed. PMID- 18380530 TI - Orthodontic treatment for an unerupted and severely rotated maxillary central incisor. A case report. AB - BACKGROUND: Many reports have indicated the treatment for permanent teeth with disturbed eruption due to the presence of supernumerary teeth. However, successful treatment for an unerupted maxillary incisor with severe rotation and another maxillary incisor with disturbed eruption by a supernumerary tooth is quite rare. CASE REPORT: We treated a 7-year-old Japanese male with an unerupted and severely rotated maxillary right incisor, along with a maxillary left incisor due to interference with eruption by an inverted supernumerary tooth. Orthodontic treatment using surgical exposure, traction and guidance of the rotated right incisor, and maxillary expansion following extraction of the supernumerary tooth were performed. FOLLOW-UP: At 27 months following extraction of the supernumerary tooth, and comprehensive orthodontic treatment led the malposititioned central incisors to a functional position and stable occlusion. PMID- 18380531 TI - Antioxidant and vascular effects of gliclazide in type 2 diabetic rats fed high fat diet. AB - Diabetes mellitus is characterized by oxidative stress, which in turn determines endothelial dysfunction. Gliclazide is a sulphonylurea antidiabetic drug with antioxidant effects due to its azabicyclo-octyl ring. It has been reported to potentially protect the vasculature through improvements in plasma lipid levels and platelet function. We hypothesized that gliclazide has a beneficial effect on endothelial function in Goto-Kakizaki rats (GK), an animal model of type 2 diabetes fed an atherogenic diet for 4 months. We evaluated the influence of gliclazide on both metabolic and oxidative status and NO-mediated vasodilation. GKAD rats showed increased oxidative stress and impaired endothelium-dependent vasodilation. GKAD rats treated with gliclazide showed increased sensitivity to NO-mediated vasodilation, a significant decrease in fasting glycemia and insulinemia, and a significant decrease in systemic oxidative stress. In conclusion, our results suggest that gliclazide treatment improves NO-mediated vasodilation in diabetic GK rats with dyslipidemia probably due to its antioxidant effects, although we cannot rule out substantial benefits due to a reduction in fasting blood glucose. The availability of a compound that simultaneously decreases hyperglycemia, hyperinsulinemia, and inhibits oxidative stress is a promising therapeutic candidate for the prevention of vascular complications of diabetes. PMID- 18380532 TI - Rapid but not slow spinal cord compression elicits neurogenic pulmonary edema in the rat. AB - The development of neurogenic pulmonary edema (NPE) can be elicited by an immediate epidural balloon compression of the thoracic spinal cord. To evaluate whether a slower balloon inflation could prevent NPE development, we examined the extent of NPE in animals lesioned with a rapid (5 microl - 5 microl - 5 microl) or slow rate (3 microl - 2 microl - 2 microl - 2 microl - 2 microl - 2 microl - 2 microl) of balloon inflation. These groups were compared with the NPE model (immediate inflation to 15 microl) and with healthy controls. Slow balloon inflation prevented NPE development, whereas the pulmonary index and histology revealed a massive pulmonary edema in the group with a rapid rate of balloon inflation. Pulmonary edema was preceded by a considerable decrease in heart rate during the inflation procedure. Moreover, rapid inflation of balloon in spinal channel to either 5 microl or 10 microl did not cause NPE. Thus, a slow rate of balloon inflation in the thoracic epidural space prevents the development of neurogenic pulmonary edema, most likely due to the better adaptation of the organism to acute circulatory changes (rapid elevation of systemic blood pressure accompanied by profound heart rate reduction) during the longer balloon inflation period. It should be noted that spinal cord transection at the same level did not cause neurogenic pulmonary edema. PMID- 18380534 TI - Natriuretic peptides - physiology, pathophysiology and clinical use in heart failure. AB - The natriuretic peptides - atrial, brain and C-type - were discovered during the last twenty years. Their effects on cardiovascular, renal, cerebral and other tissues through guanylyl cyclase were uncovered. Over the past decade natriuretic peptides (NPs) became a very useful tool in the management of heart failure patients. Results of many clinical trials have shown that BNP and NT-proBNP are helpful for diagnosis of heart failure. They are also independent markers of prognosis not only in heart failure patients but also in patients with other cardiovascular diseases. Recently published data document the utility of NPs in guiding treatment of heart failure patients. In this article, we focus on basic biochemical and physiological characteristics of NPs as well as on their significance in management of heart failure patients. Some limitations and pitfalls of NPs levels interpretation in diagnosing heart failure are also discussed. PMID- 18380533 TI - Differential expression of VEGFA, TIE2, and ANG2 but not ADAMTS1 in rat mesenteric microvascular arteries and veins. AB - Microvessels respond to metabolic stimuli (e.g. pO(2)) and hemodynamic forces (e.g. shear stress and wall stress) with structural adaptations including angiogenesis, remodeling and pruning. These responses could be mediated by differential gene expression in endothelial and smooth muscle cells. Therefore, rat mesenteric arteries and veins were excised by microsurgery, and mRNA expression of four angioadaptation-related genes was quantified by real time duplex RT-PCR in equal amounts of total RNA, correlated to two different house keeping genes (beta-actin, GAPDH). The results show higher expression of VEGFA, TIE2, and ANG2 in arteries than in veins, but equal expression of ADAMTS1. Higher availability of VEGFA mRNA in endothelial cells of arteries shown here could contribute to the maintenance of mechanically stressed blood vessels and counteract pressure-induced vasoconstriction. PMID- 18380535 TI - Testosterone modulates cardiomyocyte Ca(2+) handling and contractile function. AB - The extent to which sex differences in cardiac function may be attributed to the direct myocardial influence of testosterone is unclear. In this study the effects of gonadal testosterone withdrawal (GDX) and replacement (GDX+T) in rats, on cardiomyocyte shortening and intracellular Ca(2+) handling was investigated (0.5 Hz, 25 oC). At all extracellular [Ca(2+)] tested (0.5-2.0 mM), the Ca(2+) transient amplitude was significantly reduced (by approximately 50 %) in myocytes of GDX rats two weeks post-gonadectomy. The time course of Ca(2+) transient decay was significantly prolonged in GDX myocytes (tau, 455+/-80 ms) compared with intact (279+/-23 ms) and GDX+T (277+/-19 ms). Maximum shortening of GDX myocytes was markedly reduced (by more than 60 %) and relaxation significantly delayed (by more than 35 %) compared with intact and GDX+T groups. Thus testosterone replacement completely reversed the cardiomyocyte hypocontractility induced by gonadectomy. These results provide direct evidence for a role of testosterone in regulating functional Ca(2+) handling and contractility in the heart. PMID- 18380536 TI - Evaluation of oxidative status in acetaminophen treated rat hepatocytes in culture. AB - The present study describes the estimation of acetaminophen (AAP) toxicity in cultured rat hepatocytes. We used different concentrations of AAP - 1, 2.5, 5, 10 and 20 mM, to test influence of AAP on cellular viability, functional capacity and oxidative status at given time intervals. WST 1 test showed decrease of dehydrogenase activity in 5, 10 and 20 mM AAP to 75 % of control values after 1 hour of incubation. At 12 h of treatment, all AAP concentrations decreased WST-1 signal; no enzyme activity was found since 18 h in cells treated with 20 mM AAP according to LDH leakage test performed at 24 h of incubation. Functional capacity was tested by albumin assay where the decrease was strictly related to AAP dose. Intracellular oxidative status was assessed by analysis of GSH/GSSG levels and time course of ROS production and glutathione reductase (GR) activity. Increased ROS production was found already after 3 h of incubation in 2.5, 5, 10 and 20 mM AAP, respectively. The highest ROS production was measured after 12 h treatment. GR activity was decreased already after 3 h of incubation and remained also decreased in cells treated with 2.5, 5, 10 and 20 mM AAP during further incubation. PMID- 18380538 TI - Lower wall shear rate of the common carotid artery in treated type 2 diabetes mellitus with metabolic syndrome. AB - Arterial sites with low wall shear stress (WSS) are more prone to the development of atherosclerotic plaques, as was observed in carotid arteries in subjects with atherosclerosis risk factors. Type 2 diabetes mellitus (DM), hypertension, hyperlipidemia and other components of the metabolic syndrome, are associated with high risk for symptomatic cerebrovascular disease. It was shown by others that untreated type 2 DM is associated with lower WSS in common carotid arteries. However, the cardiovascular risk of type 2 DM could be modified by therapy. The aim of our study was to test the hypothesis that treated type 2 DM subjects with metabolic syndrome still have lower WSS in common carotid arteries than healthy controls. We enrolled 26 compensated DM subjects with metabolic syndrome, treated by metformin, statins and ACEI for more than 6 months, and 22 aged-comparable healthy controls. Wall shear rate (WSR) was used as a measure of WSS. A linear 3 11 MHz probe was used to measure blood velocity and internal diameter in the common carotid arteries. We compared observed values of WSR adjusted for age by ANCOVA. Wall shear rate was significantly lower in DM group than in control subjects: peak (systolic) values of wall shear rate were 410+/-130 s(-1) vs. 487+/-111 s(-1) (p<0.005). DM subjects had significantly lower WSR, because of both thinner lumen and slower blood flow velocities. Lower WSR was accompanied by higher IMT (0.73+/-0.12 mm vs. 0.64+/-0.11 mm, p<0.001). Treated subjects with compensated type 2 DM with metabolic syndrome still have atherogenic hemodynamic profile. These findings might help to understand faster progression of atherosclerosis in diabetic subjects with metabolic syndrome despite up-to-date medication. PMID- 18380537 TI - A non-invasive method for detecting the metabolic stress response in rodents: characterization and disruption of the circadian corticosterone rhythm. AB - Plasma corticosterone (CORT) measures are a common procedure to detect stress responses in rodents. However, the procedure is invasive and can influence CORT levels, making it less than ideal for monitoring CORT circadian rhythms. In the current paper, we examined the applicability of a non-invasive fecal CORT metabolite measure to assess the circadian rhythm. We compared fecal CORT metabolite levels to circulating CORT levels, and analyzed change in the fecal circadian rhythm following an acute stressor (i.e. blood sampling by tail veil catheter). Fecal and blood samples were collected from male adolescent rats and analyzed for CORT metabolites and circulating CORT respectively. Fecal samples were collected hourly for 24 h before and after blood draw. On average, peak fecal CORT metabolite values occurred 7-9 h after the plasma CORT peak and time matched fecal CORT values were well correlated with plasma CORT. As a result of the rapid blood draw, fecal production and CORT levels were altered the next day. These results indicate fecal CORT metabolite measures can be used to assess conditions that disrupt the circadian CORT rhythm, and provide a method to measure long-term changes in CORT production. This can benefit research that requires long-term glucocorticoid assessment (e.g. stress mechanisms underlying health). PMID- 18380539 TI - Asymmetric dimethylarginine and the effect of folate substitution in children with familial hypercholesterolemia and diabetes mellitus type 1. AB - A recently discussed cardiovascular risk factor, asymmetric dimethylarginine (ADMA), is known to act as an endogenous inhibitor of endothelial nitric oxide synthase. The aim of this study was to establish 1) the relationship between ADMA and ultrasonographically or biochemically determined endothelial dysfunction in children, and 2) the effect of folate supplementation on these parameters. The study cohort included 32 children with familial hypercholesterolemia (FH), 30 with diabetes mellitus type 1 (DM1) and 30 age-matched healthy children as the control group. Furthermore, twenty-eight randomly selected FH and DM1 children were re-examined after 3-months supplementation with folic acid. Baseline levels of ADMA and oxidized low density lipoproteins (oxLDL) were significantly higher in FH group than in DM1 and healthy children. Children in DM1 group had significantly lower concentration of homocysteine, but ADMA levels were normal. Folic acid supplementation significantly lowered homocysteine and hsCRP levels in both FH and DM1 group; however, ADMA and oxLDL concentrations remained unaltered. In conclusion, ADMA and oxLDL appear to be associated with endothelial dysfunction in children with FH. Administration of folic acid did not influence these markers in both FH and DM1 children. PMID- 18380540 TI - The role of ghrelin in the regulation of food intake in patients with obesity and anorexia nervosa. AB - Gastrointestinal hormones play an important role in the neuroendocrine regulation of food intake and postprandial satiety. Ghrelin is a 28-amino acid orexigenic peptide produced mainly by the stomach that is involved in both the long-term regulation of body weight and the short-term regulation of postprandial satiety. Impairments in ghrelin secretion may in concert with other factors play an important role in the development of both obesity and anorexia nervosa. Despite an intensive research the critical factors regulating physiological postprandial ghrelin response in healthy individuals and its modification by the presence of obesity and anorexia nervosa are only partially understood. The potential contribution of ghrelin to the differences of diet- vs. surgical-induced weight losses in morbidly obese patients is now also being recognized. The aim of this review is to summarize the current knowledge about the physiology and pathophysiology of ghrelin and to discuss its potential in the prevention and/or treatment of obesity and anorexia nervosa. PMID- 18380541 TI - Diabetes and thyroid hormones affect connexin-43 and PKC-epsilon expression in rat heart atria. AB - We have examined the changes of intercellular electrical coupling protein connexin-43 (Cx43) and of PKC-epsilon in heart atria of diabetic rats and/or after the treatment with triiodothyronine (T(3)). Diabetes was induced in Wistar Kyoto rats by streptozotocin (50 mg/kg, i.v.) and atria were examined after 5 (acute stage) and 10 (chronic stage) weeks. T(3) (10 microg/100 g/day) was applied via a gastric tube for the last 10 days prior to the end of the experiments to non-diabetic and to the half of diabetic rats. Expression and phosphorylated status of Cx43, as well as expression of PKC-epsilon, were analyzed by Western blots using mouse monoclonal anti-Cx43 and rabbit polyclonal anti-PKC-epsilon antibodies. We found that the Cx43 expression was significantly increased after the treatment with T(3) and in the acute diabetes. Both in diabetes and after T(3) treatment the phosphorylation of Cx43 isoforms was markedly suppressed compared to the non-diabetic and T(3)-untreated controls. Such a down-regulation was less pronounced in diabetic rats after the T(3) treatment. The expression of atrial PKC-epsilon was increased in diabetic rats. This increase was suppressed after T(3) administration and the expression was decreased in T(3)-treated non-diabetic rats. We suggest that the reduced Cx43 phosphorylation in diabetic and hyperthyroid rats can deteriorate a cell-to-cell coupling and consequently facilitate a development of atrial tachyarrhythmia in diabetic or hyperthyroid animals. PMID- 18380542 TI - Exercise-induced prostacyclin release positively correlates with VO(2max) in young healthy men. AB - In this study we have evaluated the effect of maximal incremental cycling exercise (IE) on the systemic release of prostacyclin (PGI(2)), assessed as plasma 6-keto-PGF(1alpha) concentration in young healthy men. Eleven physically active - untrained men (mean +/- S.D.) aged 22.7 +/- 2.1 years; body mass 76.3 +/ 9.1 kg; BMI 23.30 +/- 2.18 kg . m(-2); maximal oxygen uptake (VO(2max)) 46.5 +/- 3.9 ml . kg(-1) . min(-1), performed an IE test until exhaustion. Plasma concentrations of 6-keto-PGF(1alpha), lactate, and cytokines were measured in venous blood samples taken prior to the exercise and at the exhaustion. The net exercise-induced increase in 6-keto-PGF(1alpha) concentration, expressed as the difference between the end-exercise minus pre-exercise concentration positively correlated with VO(2max) (r=0.78, p=0.004) as well as with the net VO(2) increase at exhaustion (r=0.81, p=0.003), but not with other respiratory, cardiac, metabolic or inflammatory parameters of the exercise (minute ventilation, heart rate, plasma lactate, IL-6 or TNF-alpha concentrations). The exercise-induced increase in 6-keto-PGF(1alpha) concentration?? was significantly higher (p=0.008) in a group of subjects (n=5) with the highest VO(2max) when compared to the group of subjects with the lowest VO(2max), in which no increase in 6-keto-PGF(1alpha) concentration was found. In conclusion, we demonstrated, to our knowledge for the first time, that exercise-induced release of PGI(2) in young healthy men correlates with VO(2max), suggesting that vascular capacity to release PGI(2) in response to physical exercise represents an important factor characterizing exercise tolerance. Moreover, we postulate that the impairment of exercise induced release of PGI(2) leads to the increased cardiovascular hazard of vigorous exercise. PMID- 18380543 TI - Algesthesia after epileptic seizure. AB - The consequences of epileptic seizures related to postictal inhibition in early postictal period include postictal analgesia. We studied this phenomenon over 96 h following flurothyl-induced seizures in adult male Wistar rats. Nociception of control (no seizure) and seizured groups were tested using the plantar and von Frey hair tests. We determined latency of forepaw and hind paw reactions using plantar tests and the number of von Frey hairs reactions. Shortly after seizures, longer plantar test latencies were seen relative to the control group. Before the seizures the plantar test reaction times were significantly shorter in forepaws than in hind paws. The effect disappeared post-seizure and surprisingly, it also disappeared at the corresponding time in controls; it reappeared after 48 h in the seizure group and after 24 h in controls. Differences in the von Frey hairs test occurred at 5 and 60 min post-seizure, however, these differences could not be explained by limb anatomy; although, different thermal and mechanical nociception mechanisms could be significant. The unexpected reactions in controls could be related to brief social and physical interactions between the two groups. PMID- 18380544 TI - Fiber type composition of unoperated rat soleus and extensor digitorum longus muscles after unilateral isotransplantation of a foreign muscle in long-term experiments. AB - We examined the effects of the unilateral heterochronous isotransplantation on the fiber type composition and myosin heavy chain (MyHC) isoform content of unoperated slow soleus and fast extensor digitorum longus muscles of female inbred Lewis strain rats. Comparison was made between "control" unoperated muscles of experimental rats (after intramuscular transplantation surgery) with the corresponding muscles of completely naive (unoperated) rats of three age groups (5-, 8- and 14-month-old). This was done in order to ascertain whether these muscles can be used as reliable controls to the transplanted and host muscles for our ongoing grafting experiments. The fiber type composition was determined by assessing the histochemical reaction for myofibrillar adenosine triphosphatase, the MyHC isoform content was determined immunocytochemically using monoclonal antibodies specific to different MyHC isoforms and by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Our experiments show that the heterochronous intramuscular isotransplantation procedure had no significant effect on the fiber type composition and MyHC isoform content of the "control" unoperated muscles of the experimental rats when compared to the corresponding muscles of the naive animals. Furthermore, the duration and type of isotransplantation did not also lead to differences among corresponding "control" muscles of experimental animals. We conclude that the unoperated muscles of the experimental rats can be used as controls in our current transplantation project dealing with long-term grafting experiments. PMID- 18380545 TI - Activation of adenosine A(3) receptors potentiates stimulatory effects of IL-3, SCF, and GM-CSF on mouse granulocyte-macrophage hematopoietic progenitor cells. AB - Adenosine A(3) receptor agonist N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA) has been tested from the point of view of potentiating the effects of hematopoietic growth factors interleukin-3 (IL-3), stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony stimulating factor (G-CSF) on the growth of hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) in suspension of normal mouse bone marrow cells in vitro. IB-MECA alone induced no GM-CFC growth. Significant elevation of numbers of GM-CFC evoked by the combinations of IB-MECA with IL-3, SCF, or GM-CSF as compared with these growth factors alone has been noted. Combination of IB MECA with G-CSF did not induce significantly higher numbers of GM-CFC in comparison with G-CSF alone. Joint action of three drugs, namely of IB-MECA + IL 3 + GM-CSF, produced significantly higher numbers of GM-CFC in comparison with the combinations of IB-MECA + IL-3, IB-MECA + GM-CSF, or IL-3 + GM-CSF. These results give evidence of a significant role of selective activation of adenosine A(3) receptors in stimulation of the growth of granulocyte/ macrophage hematopoietic progenitor cells. PMID- 18380546 TI - Correlation of ultrastructural changes of endothelial cells and astrocytes occurring during blood brain barrier damage after traumatic brain injury with biochemical markers of BBB leakage and inflammatory response. AB - Focal cerebral contusion can be dynamic and expansive. It has been proved that subsequent expansive contusion is caused by brain parenchyma damage, especially BBB damage. We investigated a group of patients with traumatic brain injury. The patients (n=18) were divided into group I (n=7) of patients submitted to neurosurgery due to expansive contusion, and group II (n=11) of patients without surgery. Serum concentrations of NSE and S-100B protein were measured by electrochemiluminescence immunoassay, interleukin-6 (IL-6) was measured by chemiluminescent sequential immunometric assay and matrix metalloproteinases (MMP 9, MMP-2) were measured by immunoassays. Cortical biopsy specimens of brain were investigated by electron microscopy in patients with trauma brain injury submitted to neurosurgery. Biochemical investigation from first day up to third day after traumatic brain injury proved increased values of IL-6 (302.2+/-119.9 vs. 59.6+/-11.9 ng/l, p<0.02) and S-100B protein (3.064+/-1.064 vs. 0.649+/-0.182 microg/l, p<0.05) in patients with expansive lesion compared to patients without expansive contusion. Significantly higher levels of MMP-9 (150.4+/-28.46 vs. 74.11+/-13.16 ng/l, p<0.05) and of MMP-2 (814.5+/-126.3 vs. 523.1+/-25.28 ng/l, p<0.05) were found during first 3 days after admission in group I compared to group II. MMP-9 has also elevated in group II from lower values after admission (74.11+/-13.16 ng/l) up to high levels on the 10th day of hospitalization (225.1+/-49.35 ng/l). Ultrastructural investigation of endothelial cells and surrounded tissue revealed perivascular hemorrhage, increased pinocytic activity of endothelial cells, and cytotoxic edema of astroglial cells. Multivesical bodies were disclosed inside the endothelial cells. Higher levels of serum protein S-100B and IL-6 correlated with ultrastructural changes of endothelial cells, and with inflammatory response following TBI, respectively. PMID- 18380548 TI - Re: implant placement with or without simultaneous tooth extraction in patients taking oral bisphosphonates: postoperative healing, early follow-up, and the incidence of complications in two private practices. Fugazzoto PA, Lightfoot WS, Jaffin R, Kumar A.(J Periodontol 2007; 78: 1664-1669). PMID- 18380550 TI - Free gingival grafts to increase keratinized tissue: a retrospective long-term evaluation (10 to 25 years) of outcomes. AB - BACKGROUND: Gingival augmentation procedures are indicated primarily to increase an insufficient amount of gingiva and sometimes to halt the progression of gingival recession. The aim of this retrospective long-term study was to evaluate changes in the amount of keratinized tissue (KT) and in the position of the gingival margin after free gingival graft procedures over a period of 10 to 25 years. METHODS: One hundred three subjects presenting with 224 sites completely lacking attached gingiva associated with gingival recessions were treated in a private practice. The experimental sites were treated with gingival augmentation procedures (free gingival grafts). The grafts were positioned at the presurgical level of the gingival margin or in a submarginal position. Clinical variables, including recession depth, amount of KT, and probing depth (PD), were measured at baseline (T(0)), 1 year after surgery (T(1)), and at the end of the follow-up period (10 to 25 years) (T(2)) and analyzed using descriptive statistics and multilevel models. RESULTS: From T(0) to T(1), the gingival margin shifted coronally 0.8 mm, and KT increased 4.2 mm. From T(1) to T(2), the gingival margin shifted coronally 0.6 mm, and the overall KT decreased 0.7 mm. PD remained stable. CONCLUSION: Gingival augmentation procedures performed in sites with an absence of attached gingiva associated with recessions provide an increased amount of KT associated with recession reduction over a long period of time. PMID- 18380551 TI - 5-year longitudinal study of survival rate and periodontal parameter changes at sites of maxillary canine autotransplantation. AB - BACKGROUND: Although dental transplantation is an accepted treatment modality, few studies have evaluated the prognosis for autotransplantation of maxillary canines, and changes in clinical periodontal parameters still need to be demonstrated. The objectives of this long-term study were to evaluate the prognosis and changes in clinical attachment level (CAL), probing depth (PD), soft tissue recession (REC), and radiographic bone level changes between years 1 and 5 for autotransplanted maxillary canines. METHODS: Thirty subjects with 32 impacted canines were scheduled for transplantation. Transplanted teeth were splinted for 4 weeks, and endodontic treatments were performed within 1 year after the operation. Radiographs were taken, and PD and CAL were measured for up to 5 years. Progressive root resorption was observed in two teeth, and one tooth was extracted during year 4. RESULTS: The survival rate was 93.5%. CAL and PD tended to increase slightly at all sites, especially at palatal sites after transplantation, whereas the greatest CAL and PD were found at approximal sites, and REC was the same as natural teeth. CONCLUSIONS: Maxillary canine transplantation can be a reliable treatment alternative. Clinical measurements demonstrated long-term stability. Based on the literature and the present findings, maxillary canine transplantation can be suggested as a promising treatment modality. PMID- 18380552 TI - Assessment of parameters affecting the presence of the central papilla using a non-invasive radiographic method. AB - BACKGROUND: This study was performed to assess the relationship between radiopaque landmarks and central papilla presence and height. The probability of the presence of a central papilla has been related to the bone crest-contact point distance. Papilla height has been measured by inserting a dental probe under local anesthesia, but this method is invasive. It would be desirable to assess the influence of other dental landmarks on the presence and height of the papilla and to use a non-invasive method for measurement of papilla height that is simple, accurate, and accepted readily by patients. METHODS: Periapical radiographs were obtained in 310 adults with fully erupted permanent dentition, healthy gingiva (plaque and gingival indexes of 0 to 1), and well-aligned maxillary central incisors. Semisoft radiopaque material was placed to fill the space coronal to the papilla tip, which enabled it to be visualized without probing. Vertical and horizontal distances were measured with an electric ruler. RESULTS: The presence of a full central papilla was related significantly to age, papillary height, and distances from the bone crest to the contact point, the buccal cemento-enamel junction, and the proximal cemento-enamel junction. When confounding factors were controlled using multifactorial logistic regression, the distance from the bone crest to the proximal cemento-enamel junction was the strongest determinant of papilla presence. CONCLUSIONS: Standard periapical radiography is a non-invasive method that can be used to study determinants of the presence of a central papilla. The factors linked to papilla recession were age, bone crest-contact point distance, bone crest-proximal cemento-enamel junction distance, and papilla height. Bone crest-proximal cemento-enamel junction distance, the strongest predictor of the probability of papilla recession, might be used to predict this probability after prosthetic or orthodontic interventions. PMID- 18380553 TI - Periodontal attachment loss in an untreated isolated population of Brazil. AB - BACKGROUND: The aim of this study was to assess the prevalence, extent, and severity of clinical attachment loss (CAL) and to investigate the association of demographic, socioeconomic, and behavioral risk indicators with CAL in an untreated isolated population in Brazil. METHODS: All subjects aged > or =12 years were identified by a census. Participants were submitted to a full-mouth clinical examination of six sites per tooth and were interviewed using a structured, written questionnaire. RESULTS: Among the 214 subjects who were interviewed and examined clinically, CAL > or =5 mm in at least one site was observed in 8% of the 12- to 19-year-olds and in all dentate subjects > or =50 years of age; the age-dependent prevalence of CAL > or =7 mm in at least one site ranged from 5% among 12- to 19-year-olds to 83% among subjects > or =50 years old. Multivariate analysis identified plaque (odds ratio [OR] = 2.8), supragingival calculus (OR = 2.9 to 10.6), age > or =30 years (OR = 11.4), and smoking (OR = 2.4) as risk indicators for CAL > or =5 mm and smoking (OR = 8.2) as a risk indicator for CAL > or =7 mm. CONCLUSIONS: CAL is highly prevalent in this isolated population. The high occurrence of CAL in young age groups and the confirmation of traditional risk indicators for CAL in this study suggest that other factors, such as host susceptibility, may be needed to explain the high levels of CAL found. Age and behavioral factors were risk indicators associated significantly with the CAL found in this population and may be useful indicators of high-risk subjects for periodontal diseases. PMID- 18380554 TI - Coronally positioned flap plus resin-modified glass ionomer restoration for the treatment of gingival recession associated with non-carious cervical lesions: a randomized controlled clinical trial. AB - BACKGROUND: The aim of this randomized clinical trial was to evaluate the treatment of gingival recession associated with non-carious cervical lesions by a coronally advanced flap alone (CAF) or in combination with a resin-modified glass ionomer restoration (CAF+R). METHODS: Nineteen subjects with bilateral Miller Class I buccal gingival recessions associated with non-carious cervical lesions were selected. The recessions were assigned randomly to receive CAF or CAF+R. Bleeding on probing (BOP), probing depth (PD), relative gingival recession (RGR), clinical attachment level (CAL), non-carious cervical lesion height (CLH), and dentin sensitivity (DS) were measured at baseline; 45 days; and 2, 3, and 6 months postoperatively. Keratinized tissue width (KTW) and keratinized tissue thickness (KTT) were measured at baseline and 6 months. The height of the non carious cervical lesion located on the root and crown were estimated, allowing calculation of root coverage. RESULTS: Both groups showed statistically significant gains in CAL and soft tissue coverage. The differences between groups were not statistically significant for BOP, PD, RGR, CAL, KTW, and KTT after 6 months. The percentages of CLH covered were 56.14% +/- 11.74% for CAF+R and 59.78% +/- 11.11% for CAF (P >0.05). The root and crown surfaces affected by the non-carious cervical lesion were 1.67 +/- 0.31 mm and 0.96 +/- 0.29 mm, respectively, for CAF+R and 1.59 +/- 0.37 mm and 1.01 +/- 0.33 mm, respectively, for CAF. The estimated root coverage was 88.02% +/- 19.45% for CAF+R and 97.48% +/- 15.36% for CAF (P >0.05). CAF+R reduced DS significantly compared to CAF (P <0.05). CONCLUSIONS: Both procedures provided similar soft tissue coverage after 6 months. Despite the fact that a greater reduction in DS was observed after CAF+R, longitudinal observations are necessary to confirm these results. PMID- 18380555 TI - Periodontal disease in Hispanic Americans with type 2 diabetes. AB - BACKGROUND: Diabetes is a major risk factor for the development of periodontal disease in certain populations. The prevalence of type 2 diabetes is increased in Hispanic Americans, but its impact on the extent and severity of periodontal disease in this population has not been determined. METHODS: Sixty-three Hispanic Americans, aged 33 to 72 years, from South Texas were grouped based on the presence or absence of type 2 diabetes. Past medical histories, including smoking, were obtained. Periodontal status was evaluated by measuring probing depth (PD), clinical attachment level (CAL), plaque, bleeding on probing, visual gingival inflammation, and calculus. RESULTS: Type 2 diabetes was associated frequently with major medical complications in this population. Diabetes was associated with significantly more calculus formation and tooth loss and an increased extent and severity of periodontitis. Subjects with diabetes had nearly three times the mean CAL and frequency of PD >6 mm than subjects without diabetes and nearly twice the frequency of moderate to advanced attachment loss (> or =3 mm). Smoking and diabetes had significant independent effects on mean CAL and the frequency of deep pockets. Diabetes and smoking combined were associated with a significantly higher frequency of sites with CAL > or =3 mm compared to healthy non-smokers, healthy smokers, and non-smokers with diabetes. CONCLUSIONS: Hispanic Americans with type 2 diabetes had more supra- and subgingival calculus, an increased extent and severity of periodontal destruction, and an increased frequency of tooth loss due to periodontitis. An additive/synergistic contribution of type 2 diabetes and smoking for increasing the extent of periodontal disease was observed. PMID- 18380556 TI - Microbiologic and immunologic characteristics of periodontal disease in Hispanic americans with type 2 diabetes. AB - BACKGROUND: The microbiology of periodontitis in type 1 diabetes has been reported, but less is known about type 2 diabetes. Moreover, these data have not linked microbial colonization, host response, and clinical presentation in type 1 or type 2 diabetes. The objectives of this study were to relate periodontal status, periodontal microorganisms, and host-response characteristics in Hispanic Americans with type 2 diabetes. METHODS: Plaque and serum samples were obtained from 63 Hispanic American subjects with and without type 2 diabetes. The microbiology of subgingival plaque samples was evaluated using DNA checkerboard hybridization, and serum antibody to a battery of oral microorganisms was determined using an enzyme-linked immunosorbent assay. RESULTS: In general, similar pathogens were present in periodontitis sites from subjects with and without type 2 diabetes, although the periodontitis sites in diabetes showed a higher frequency of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), and Campylobacter spp. Serum antibody to Campylobacter rectus was elevated in type 2 diabetes, whereas antibody to P. gingivalis and C. rectus were elevated in subjects with periodontitis, irrespective of diabetes status. Stratification of the population based upon antibody to P. gingivalis or C. rectus suggested a linkage between elevated antibody to P. gingivalis, increased frequency of diabetes, and significantly worse periodontitis. CONCLUSION: The increased severity of periodontal disease with type 2 diabetes may reflect an alteration of the pathogenic potential of periodontal bacteria and/or a modification of the characteristics of the host's inflammatory response that may contribute to a breakdown in the homeostasis of the periodontium. PMID- 18380557 TI - Root coverage outcome may be affected by heavy smoking: a 2-year follow-up study. AB - BACKGROUND: Although subepithelial connective tissue graft (CTG) has been reported to be a predictable procedure for root coverage, the impact of smoking on the long-term outcome of periodontal plastic surgery is unclear. Hence, the aim of this study was to evaluate the effect of smoking, on a long-term basis, on the stability of gingival tissue following CTG treatment of gingival recession. METHODS: Twenty-two defects were treated by CTG in canine and premolar Miller Class I and II gingival recessions (11 smokers and 11 non-smokers). The following clinical measurements were obtained at baseline and at 1, 2, 3, 4, 6, 12, 18, and 24 months after surgery: plaque and gingival indexes, extension of gingival recession (GR), probing depth (PD), clinical attachment level (CAL), and gingival thickness. Individuals smoking > or =20 cigarettes/day for > or =5 years were considered smokers. RESULTS: Data analysis demonstrated that both groups presented similar plaque and gingival indexes (P >0.05), and an intragroup analysis showed that CTG was able to promote root coverage and increase gingival thickness in both groups over time (P <0.05). However, at 24 months postoperatively, statistical analysis showed that smokers presented poorer outcomes with regard to PD, GR, and CAL (P <0.05); in addition, a more satisfactory stabilization of the gingival tissue was found in the non-smoker group. CONCLUSION: Smoking may represent a challenge to root coverage outcome for CTG because smoking significantly affected the stability of gingival tissue over time. PMID- 18380558 TI - Root proximity as a risk factor for progression of alveolar bone loss: the Veterans Affairs Dental Longitudinal Study. AB - BACKGROUND: The purpose of the present longitudinal study was to evaluate the association between root proximity and the risk for alveolar bone loss (ABL). METHODS: We used data from the Veterans Affairs Dental Longitudinal Study, a closed-panel longitudinal cohort study of 1,231 men enrolled in 1968 with triennial follow-up examinations. Periapical radiographs of mandibular incisors from subjects with > or =10 years of follow-up were selected. Interradicular distance (IRD) at the cemento-enamel junction and alveolar bone levels at baseline and last follow-up were measured using digitized radiographs. The rate of progressive ABL was determined and expressed as millimeters per 10 years. Site specific multivariate regression models were fit to evaluate the association between IRD and ABL rate, adjusting for age and smoking. Empirical standard errors and generalized estimating equations were used to account for the correlation among sites within subjects. RESULTS: There were 473 dentate subjects, aged 28 to 71 years at baseline, with > or =10 years of follow-up data available for analyses. The mean follow-up time was 23 years. The mean IRD was 1.0 +/- 0.3 mm, and the mean ABL rate during 10 years was 0.61 +/- 0.59 mm. There was a significant non-linear association between IRD and ABL rate (P <0.005). Compared to sites with IRD > or =0.8 mm, sites with IRD <0.6 mm were 28% (95% confidence interval [CI]: 11% to 48%) more likely to lose > or =0.5 mm of bone during 10 years (relative risk: 1.28 [95% CI: 1.11 to 1.48]) and 56% (95% CI: 11% to 117%) more likely to lose > or =1.0 mm of bone during 10 years (relative risk: 1.56 [95% CI: 1.11 to 2.17]). CONCLUSIONS: IRD <0.8 mm is a significant local risk factor for alveolar bone loss in mandibular anterior teeth. Measurement of IRD may have important prognostic value in making treatment decisions. PMID- 18380559 TI - Effect of platelet-rich plasma on the healing of intrabony defects treated with Beta tricalcium phosphate and expanded polytetrafluoroethylene membranes. AB - BACKGROUND: Regenerative periodontal therapy using platelet-rich plasma (PRP) and different types of bone substitutes with or without guided tissue regeneration (GTR) has been proposed as a modality to enhance the outcome of regenerative surgery. However, there are limited data from controlled clinical studies evaluating the effect of PRP on the healing of deep intrabony defects treated with a combination of bone substitutes and GTR. The aim of this study was to clinically evaluate the effect of PRP on the healing of deep intrabony defects treated with beta tricalcium phosphate (beta-TCP) and GTR by means of a non bioresorbable expanded polytetrafluoroethylene membrane. METHODS: Twenty-eight subjects with advanced chronic periodontal disease and displaying one intrabony defect were treated randomly with a combination of PRP + beta-TCP + GTR or beta TCP + GTR. Plaque index, gingival index, bleeding on probing, probing depth (PD), gingival recession, and clinical attachment level (CAL) were evaluated at baseline and at 1 year after treatment. CAL was the primary outcome variable. RESULTS: No differences in any of the investigated parameters were observed at baseline between the two groups. Healing was uneventful in all subjects. At 1 year after therapy, the sites treated with PRP + beta-TCP + GTR showed a reduction in mean PD from 9.1 +/- 0.6 mm to 3.3 +/- 0.5 mm (P <0.001) and a change in mean CAL from 10.1 +/- 1.3 mm to 5.7 +/- 1.1 mm (P <0.001). In the group treated with beta-TCP + GTR, mean PD was reduced from 9.0 +/- 0.8 mm to 3.6 +/- 0.9 mm (P <0.001), and the mean CAL changed from 9.9 +/- 1.0 mm to 5.9 +/- 1.2 mm (P <0.001). In both groups, all sites gained > or =3 mm of CAL. CAL gains > or =4 mm were noted in 86% (12 of 14 defects) of the cases treated with PRP + beta-TCP + GTR and in 79% (11 of 14 defects) of those treated with beta-TCP + GTR. No statistically significant differences in any of the investigated parameters were observed between the two groups at the 1-year reevaluation. CONCLUSION: At 1 year after surgery, both therapies resulted in significant PD reductions and CAL gains. PMID- 18380560 TI - Site-specific development of periodontal disease is associated with increased levels of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in subgingival plaque. AB - BACKGROUND: Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia (previously T. forsythensis), which are regarded as the principal periodontopathogenic bacteria, exist as a consortium in subgingival biofilms. We aimed to examine quantitative relationships between P. gingivalis, T. denticola, and T. forsythia in subgingival biofilms and the relationship between the quantity and prevalence of these three bacteria and site-specific periodontal health. METHODS: This study was cross-sectional. The study population consisted of 35 adult subjects who visited the Kyushu Dental College Hospital. Plaque samples were collected from 105 periodontal pocket sites. Quantitative analyses of each of the three periodontopathogenic bacteria were performed using real-time polymerase chain reaction with species-specific primers and hybridization probes. RESULTS: The plaque samples were divided into four groups based on the presence or absence of a periodontal pocket (probing depth [PD] > or =4 mm) and bleeding on probing (BOP), regardless of attachment loss. The proportions of all three target bacteria detected in samples from sites of periodontal disease (with PD and BOP) were markedly higher than those in the other sample groups. Cell numbers of P. gingivalis, T. denticola, and T. forsythia in the subgingival plaque of each sampling site were significantly mutually correlated and were increased in the plaque of sites of periodontal disease with PD > or =4 mm and BOP. CONCLUSION: The symbiotic effects of P. gingivalis, T. denticola, and T. forsythia, which coaggregate and exist concomitantly in subgingival biofilms, may be associated with the local development of periodontitis. PMID- 18380561 TI - Interleukin-6 polymorphisms are associated with pathogenic bacteria in subjects with periodontitis. AB - BACKGROUND: Growing evidence suggests that individual genetic susceptibility may influence the host's response to infections. Previously, we showed that a common variation in the interleukin (IL)-6 gene was associated with increased odds of detection of common periodontal pathogens from individuals with aggressive periodontitis. The aim of this study was to investigate the association between IL-6 polymorphisms and periodontopathogenic bacteria in a larger, ethnically mixed population of subjects with periodontitis. METHODS: Genomic DNA was extracted from 107 subjects diagnosed with severe forms of periodontitis to study a cluster of polymorphisms in inflammatory genes, including IL-6. The presence of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, and Tannerella forsythia (previously T. forsythensis) in their subgingival biofilm was determined by polymerase chain reaction analysis. Serum IL-6 and C-reactive protein (CRP) concentrations were analyzed by enzyme-linked immunosorbent assay. RESULTS: Multiple logistic regression analysis revealed that the IL-6 -6106 polymorphism was associated with the detection of A. actinomycetemcomitans (P = 0.009; odds ratio [OR] = 3.5; 95% confidence interval [CI]: 1.38 to 9.16) and the concomitant detection of A. actinomycetemcomitans and P. gingivalis (P = 0.015; OR = 3.6; 95% CI: 1.28 to 10.04). The IL-6 -174 polymorphism was associated with increased odds of the concomitant detection of A. actinomycetemcomitans and P. gingivalis (P = 0.042; OR = 2.8; 95% CI: 1.04 to 7.75). Haplotype analysis of all five IL-6 polymorphisms confirmed an association with the detection of A. actinomycetemcomitans (P = 0.046). The IL-6 -6106 polymorphism was also associated with CRP serum levels at multivariate analysis (P = 0.024). CONCLUSIONS: These findings confirm the hypothesis that complex interactions between the microbiota and host genome are at the basis of susceptibility to periodontitis. Periodontal disease may represent a useful model to study the pathways and mechanisms of host-pathogen interactions in inflammatory diseases. PMID- 18380562 TI - Renal alterations in prediabetic rats with periodontitis. AB - BACKGROUND: Periodontitis was shown to have an impact on glucose levels in prediabetic and diabetic rats. The Zucker fatty rat (ZFR) is a well-characterized model of prediabetes presenting with impaired glucose tolerance, hyperinsulinemia, dyslipidemia, and moderate hypertension. The aim of the present study was to investigate whether periodontitis influences kidney changes in ZFRs. METHODS: Male adult ZFRs (N = 19) and their lean littermates (N = 18) were studied. Periodontitis was induced with ligatures in half of the ZFRs and lean rats, whereas the other half served as controls. After 4 weeks, the rats were sacrificed, and the kidneys, liver, and heart were removed and weighed. Kidneys were evaluated histologically for glomerular volume and renal mRNA levels of vascular endothelial growth factor (VEGF), VEGF receptor 2, transforming growth factor-beta, connective tissue growth factor, collagen IValpha1, fibronectin, and nephrin. Urinary albumin excretion and creatinine clearance were also evaluated. RESULTS: In prediabetic ZFRs, periodontitis was associated with kidney hypertrophy (P = 0.03) and a tendency for increased glomerular volume (P = 0.06). In lean littermates, elevated fibronectin mRNA levels (P = 0.03) were noted in the presence of periodontitis. CONCLUSION: Our findings suggest the participation of periodontitis in the development of early renal changes in ZFRs. PMID- 18380563 TI - Efficacy of platelet-rich plasma on wound healing in rabbits. AB - BACKGROUND: The purpose of this study was to evaluate if the healing of full thickness skin wounds was accelerated by platelet-rich plasma (PRP). METHODS: Four 2.5 x 2.5-cm full-thickness skin wounds were created on the backs of 15 New Zealand white rabbits. One wound on each animal received 0.3, 0.6, or 0.9 ml PRP, and the fourth wound served as a control. Seven and eight animals were sacrificed after 1 or 2 weeks, respectively, to determine histomorphometrically the epithelialization rate, contraction rate, healing rate, tissue fill, and volume fractions of fibroblasts, neutrophils, macrophages, and blood vessels. RESULTS: Only the 0.6- and 0.9-ml groups had significantly lower contraction rates than the controls after 2 weeks (P <0.05). Although no statistically significant differences were found in other parameters between the PRP-treated wounds and the controls, the PRP treatment led to increases in average epithelialization rates and volume fraction of blood vessels at both time periods. The PRP also seemed to have the most positive effect on healing rate, tissue fill, and volume fraction of fibroblasts during week 1 compared to week 2. CONCLUSIONS: The PRP treatment enhanced healing in full-thickness wounds by reducing the contraction rate with a trend toward acceleration of the epithelial migration and the angiogenic response. Further studies with larger sample sizes should be conducted to improve statistical sensitivity. Longer time intervals and modifications of PRP volume should also be explored to evaluate the long-term efficacy of PRP on wound healing. PMID- 18380564 TI - Chronic stress may modulate periodontal disease: a study in rats. AB - BACKGROUND: The present study aimed to evaluate whether chronic stress (CS) affects ligature-induced periodontal disease and to investigate the impact of CS on the mRNA levels of interleukin (IL)-1beta, -1 receptor antagonist, -6, and 10, interferon-gamma (IFN-gamma), receptor activator of nuclear factor-kappa B ligand (RANKL), and osteoprotegerin in the gingival tissues of rats. METHODS: Sixty male Wistar rats were assigned randomly to three groups: G1 (control; non ligated sites), G2 (periodontal disease), and G3 (periodontal disease associated with restraint stress for 12 hours/day for the entire study). After 30 days, all animals were sacrificed by decapitation. Blood samples were taken, and the concentrations of corticosterone and catecholamines were measured as biomarkers of CS. Marginal tissues around ligated and non-ligated teeth were harvested, and gene expression was assessed by quantitative polymerase chain reaction. Moreover, the area of bone loss (ABL) was determined histometrically. RESULTS: Data analysis showed that CS increased serum levels of stress biomarkers (P <0.05), ligature placement resulted in a significant ABL compared to non-ligated sites, CS significantly increased the amount of ABL in inflamed sites (P <0.001), and CS significantly increased mRNA levels of proinflammatory (IL-1beta and -6 and IFN gamma) and anti-inflammatory (IL-10) cytokines and proresorptive factor (RANKL) in ligated sites (P <0.05). CONCLUSION: CS significantly increased bone loss resulting from ligature-induced periodontitis by a local increase in proinflammatory and proresorptive factors. PMID- 18380565 TI - The reproducibility of curet sampling of subgingival biofilms. AB - BACKGROUND: Because few studies have examined the critical issue of sampling reproducibility, the purpose of the present study was to examine the reproducibility of curet sampling of subgingival biofilms. METHODS: Seven subgingival biofilm samples were taken successively, using a curet, from each of 80 sites and individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization. One healthy site was sampled in each of 20 periodontally healthy subjects, and one sulcus/pocket of < or =3, 4 to 5, and > or =6 mm was sampled in each of 20 subjects with chronic periodontitis. The significance of differences in counts and proportions of individual species at the seven successive samplings for each probing depth (PD) category was determined using the Kruskal-Wallis test. The reproducibility of species proportions at each PD category was measured using the coefficient of variation (CV), and the consistency of microbial profiles across samples was examined using the minimum similarity coefficient. RESULTS: There was no significant difference in the mean proportions of the 40 test species in the seven successive samples in each of the four PD categories. The median CV for individual species in the same site was 0.79 (95% confidence interval [CI]: 0.76 to 0.82) compared to 1.76 (95% CI: 1.69 to 1.82) in samples from different sites. The within-site mean minimum similarity coefficient (+/- SEM) was 51.2% +/- 2.2%, and it was 27.9% +/- 0.3% between sites. CONCLUSION: The proportions of species remained consistent in successive curet samples, indicating that the use of curets provided a reliable and reproducible method to obtain subgingival samples. PMID- 18380566 TI - Effect of platelet-rich plasma on cell adhesion, cell migration, and myofibroblastic differentiation in human gingival fibroblasts. AB - BACKGROUND: Platelet-rich plasma is a blood-derived fraction that contains a high concentration of platelets and growth factors. It was proposed that the use of this platelet concentrate stimulates tissue repair. However, little is known about the biologic response of gingival fibroblasts to platelet's derived growth factors. In the present study, we evaluated whether platelet-rich plasma modulated cell adhesion, cell migration, and myofibroblastic differentiation in primary cultures of human gingival fibroblasts. METHODS: We studied the response of primary cultures of gingival fibroblasts to thrombin-activated platelet-rich plasma fractions. Cell adhesion was evaluated through a colorimetric assay. Cell spreading, actin cytoskeleton remodeling, and focal adhesion distribution were assessed through light and immunofluorescence microscopy. Cell migration was analyzed using a bicameral cell culture system. Smooth muscle actin production was studied through Western blotting. RESULTS: Exposure of gingival fibroblasts to platelet-rich plasma stimulated adhesion and spreading of cells on fibronectin matrices, the development of actin-enriched cellular extensions, and formation of focal adhesions. Platelet-rich plasma also promoted cell migration and invasion through a reconstituted basement membrane matrix. Differentiation into the myofibroblastic phenotype, assessed through the production of smooth muscle actin, was also stimulated by platelet-rich plasma preparations. CONCLUSION: Platelet-rich plasma may modulate several cell responses potentially involved in wound healing such as cell adhesion, cell migration, and myofibroblastic differentiation. PMID- 18380567 TI - Cellular motility of Down syndrome gingival fibroblasts is susceptible to impairment by Porphyromonas gingivalis invasion. AB - BACKGROUND: Severe periodontal breakdown is often associated with Down syndrome (DS); however, the etiology of this condition is not understood fully. Cellular motility of gingival fibroblasts is a critical event for wound healing and regeneration of periodontal tissues. Porphyromonas gingivalis is known to be a periodontal pathogen that invades host cells, contributing to periodontal destruction. In this study, we examined the influence of P. gingivalis infection on the motility of DS gingival fibroblasts (DGFs). METHODS: DGFs and normal gingival fibroblasts (NGFs) were infected with P. gingivalis with type II fimbriae, and cellular motility was evaluated using an in vitro wounding assay. Protein degradation of alpha5beta1-integrin subunits and a migration-regulating signaling molecule, paxillin, were investigated using specific antibodies. The adhesion to and invasion of fibroblasts by P. gingivalis were determined with a colony forming assay. The gene expressions of alpha5beta1-integrin subunits were also quantified using a reverse transcription-polymerase chain reaction method. RESULTS: The cellular motility of DGFs was impaired significantly by P. gingivalis compared to NGFs, and the former were invaded readily by P. gingivalis. Further, cellular paxillin from DGFs was degraded markedly by the pathogen. Although protein degradation of alpha5beta1 integrin was induced, its mRNA expression was not affected significantly. CONCLUSIONS: P. gingivalis readily invades DGFs and subsequently degrades paxillin, which impairs cellular motility and likely prevents wound healing and the regeneration of periodontal tissues. These characteristics may be involved in the etiology of DS periodontitis. PMID- 18380568 TI - Performance of threaded versus sintered porous-surfaced dental implants using open window or indirect osteotome-mediated sinus elevation: a retrospective report. AB - BACKGROUND: The purpose of this retrospective report was to evaluate the performance of dental implants placed in a teaching environment in patients requiring maxillary sinus elevation. METHODS: Threaded (acid-washed or sand blasted acid-etched) and sintered porous-surfaced (SPS) press-fit implants were used. Sinuses were managed using direct (open window) or indirect (osteotome mediated) techniques. Records were available for 97 implants in 62 patients. Preoperative subantral bone heights were determined from radiographs, primarily panoramic. Of 69 threaded implants used, 40 and 29 were placed using direct (DTH) and indirect (ITH) techniques, respectively. Twenty-eight SPS implants were placed using the indirect technique. Implant failure rates using the Kaplan-Meier method of analysis and cumulative crestal bone loss (the latter based on bone levels seen in the most recent radiographs) were determined for both types of implants. RESULTS: The mean preoperative subantral bone height for DTH implants was 5.0 mm (range, 1 to 12 mm). Preoperative bone heights for ITH implants and SPS implants placed using the indirect approach were 7.2 mm (range, 4 to 12 mm) and 4.2 mm (range, 3 to 6 mm), respectively. Significant differences in preoperative bone height were seen between DTH and ITH implants (P <0.0001) and between ITH and SPS implants (P <0.0001). Average functional times were 18.7 months and 16.3 months for DTH and ITH implants, respectively, whereas the average functional time for SPS implants was 49.9 months. Time in function was significantly greater (P <0.0001) for SPS than DTH and ITH implants. Significant differences in implant length were also seen, with SPS implants significantly shorter than DTH or ITH implants. Three implants failed to integrate, one in each of the three treatment groups, giving initial survival rates of 97.5%, 96.6%, and 96.4% for DTH, ITH, and SPS implants, respectively. Mean cumulative crestal bone loss measurements were 1.84, 1.90, and 0.57 mm for DTH, ITH, and SPS implants, respectively. Bone loss was significantly less with SPS implants than with DTH or ITH implants. A second SPS implant failed after 7 years in function, likely because of prosthetic deficiencies. This late failure reduced the Kaplan-Meier survival rate to 80.4% for the SPS group. CONCLUSION: Results from this teaching clinic suggest that the use of dental implants with sinus elevation procedures is a predictable treatment for the resorbed posterior maxilla. PMID- 18380569 TI - Implant placement at the time of mandibular molar extraction: description of technique and preliminary results of 341 cases. AB - BACKGROUND: Theoretically, the ability to place implants in ideal positions at the time of mandibular molar extraction with concomitant regenerative therapy would simplify and shorten the course of therapy for patients. METHODS: A total of 341 implants were placed in 320 individuals at the time of mandibular molar hemisection and extraction. Concomitant regenerative therapy was performed around 332 of the placed implants. No regenerative therapy was performed around the remaining nine implants. Eleven additional sites, in which simultaneous implant placement was planned, were treated instead with regenerative therapy alone using graft material and a covering membrane. Implants were placed in these sites in subsequent surgical visits. RESULTS: One implant was mobile 3 weeks postinsertion. A second implant was lost after 30 months in function. All other implants were stable at the time of uncovery 3 to 7 months postinsertion. A total of 339 implants have been in function for up to 6 years, with a mean time in function of 30.8 months, yielding a cumulative survival rate of 99.1%. CONCLUSION: Implants may be placed in ideal restorative positions at the time of mandibular molar extraction with or without concomitant regenerative therapy. PMID- 18380570 TI - Gingival metastasis from salivary duct carcinoma of the parotid gland. AB - BACKGROUND: This article reports a rare case of metastasis of salivary duct carcinoma of the parotid gland to the gingiva and reviews the occurrence of metastatic processes to the oral mucosa. METHODS: A 67-year-old white male presented with a chief complaint of a painless nodular tissue growth on the gingiva with reportedly 5 months of evolution. The intraoral examination revealed a reddish, superficially ulcerated nodular lesion ( approximately 2 cm in diameter) on the right mandibular buccal attached gingiva, and the clinical aspect was that of a benign reactive lesion. The patient had undergone a parotidectomy for removal of a salivary duct carcinoma of the parotid gland almost 1 year before. A biopsy of the gingival lesion was performed, and the biopsied tissue was forwarded for histopathologic examination. RESULTS: The analysis of the histopathologic sections of the gingival lesion revealed histomorphologic characteristics very similar to those of the primary parotid gland tumor. The definitive diagnosis was gingival metastasis from a salivary duct carcinoma of the parotid gland. The patient died of complications of a pulmonary metastasis 1 month after the diagnosis of the oral metastatic lesion. CONCLUSIONS: Gingival lesions that mimic reactive and hyperplastic lesions may be metastases from malignant neoplasias of diverse origins. An accurate and timely diagnosis is crucial to establish proper and immediate treatment of the metastatic tumor and possibly identify an occult primary malignant neoplasia. PMID- 18380571 TI - Tunnel technique with a surgical maneuver to increase the graft extension: case report with a 3-year follow-up. AB - BACKGROUND: This article reports a clinical case with a 3-year follow-up in which a subepithelial connective tissue graft (SCTG) was used with the tunnel technique to treat multiple gingival recessions, and describes a technique used to enlarge the extension of the graft. METHODS: A 41-year-old female patient was referred for evaluation and treatment of maxillary multiple recessions. Following basic therapy, the plaque index was 23%, and the gingival index was 12%. Thus, SCTG with the tunnel technique was proposed to provide root coverage of Miller Class I recession on teeth #8 through #11 and a Miller Class III recession on tooth #12. After the donor area had been prepared, SCTG was removed and split cross sectionally to lengthen it. The graft was placed through the tunnel and sutured. RESULTS: Two weeks after the surgical procedure, the tissue color was nearly homogeneous with some reddish regions where the connective tissue was left uncovered, and there were no signs of incisions or suture marks. After 3 years of follow-up, the mean coverage of the recessions was 2.2 +/- 0.7 mm (74.2%), which corresponded to the gain of keratinized tissue. In addition, a gain in tissue thickness was observed. CONCLUSION: In a long-term evaluation, the tunnel technique with the elongated SCTG was used successfully for treatment of multiple gingival recessions with an increase of the soft tissue volume and gain of keratinized tissue. PMID- 18380572 TI - Gingival enlargement as a manifestation of tuberous sclerosis: case report and periodontal management. AB - BACKGROUND: Tuberous sclerosis is an autosomal-dominant inherited disease involving many organs of the body. Oral manifestations include gingival enlargement, fibromas, and dental enamel pitting. The report presents a case of tuberous sclerosis with gingival enlargement histologically consistent with angiofibroma, describes its successful periodontal management, and reviews the literature associated with oral manifestations of tuberous sclerosis. METHODS: A 26-year-old white male presented to the Department of Periodontics and Implant Dentistry, New York University College of Dentistry, with a diagnosis of tuberous sclerosis and a chief complaint of gingival enlargement affecting mastication and esthetics. Following a complete medical history review, consultation with the patient's medical team at New York University Medical Center, and a thorough oral and periodontal examination, a treatment plan was developed that included oral hygiene instructions, mechanical debridement, and periodontal reevaluation. This was followed by gingivectomy, which provided improved function and esthetics. Excised tissue was submitted for histologic examination. The patient was followed every 2 months for assessment of the outcome of the surgical treatment. An extensive search of the dental and dermatologic literature was performed on MEDLINE. RESULTS: Histologic examination of the gingival tissue revealed features consistent with angiofibroma. Fifteen months following gingivectomy, the contours and gingival surface appearance remained normal. CONCLUSIONS: The gingival enlargement was histologically consistent with the characteristic angiofibromas of tuberous sclerosis. The gingival enlargement responded very well to gingivectomy and periodontal maintenance. PMID- 18380573 TI - Periodontal manifestation of leukocyte adhesion deficiency type I. AB - BACKGROUND: Leukocyte adhesion deficiency type I (LAD-I) is an autosomal recessive immunodeficiency disorder characterized by defects in the integrin receptors of white blood cells that lead to impaired adhesion and chemotaxis. Affected patients are susceptible to recurrent bacterial and fungal infections, impaired pus formation, delayed wound healing, and periodontitis. METHODS: A case of generalized advanced periodontal destruction of the permanent and deciduous dentition in a young Jordanian girl with a severe phenotype of LAD-I is presented in this report. The medical diagnosis was made based on the characteristic clinical and laboratory findings, in particular the total absence of CD18, CD11b, and CD11c as determined by flow cytometry sorting. RESULTS: Periodontal findings in this patient include the early onset of the disease, which affected the primary teeth and permanent dentitions, the intense redness and inflammation of the gingiva, and the rapid periodontal destruction that seems refractory to conventional non-surgical periodontal therapy. CONCLUSION: This report emphasizes the importance of the differential diagnosis of severe immunodeficiency disorders in children and adolescents and mandates the importance of combined care by medical and dental practitioners to prevent tooth loss and control oral infection. PMID- 18380575 TI - Evaluation of induction characteristics and hypnotic potency of isoflurane and sevoflurane in healthy dogs. AB - OBJECTIVE: To determine induction characteristics and the minimum alveolar concentration (MAC) at which consciousness returned (MACawake) in dogs anesthetized with isoflurane or sevoflurane. ANIMALS: 20 sexually intact male Beagles. PROCEDURES: In experiment 1, 20 dogs were randomly assigned to have anesthesia induced and maintained with isoflurane or sevoflurane. The MAC at which each dog awoke in response to auditory stimulation (MACawake-noise) was determined by decreasing the end-tidal concentration by 0.1 volume (vol %) every 15 minutes and delivering a standard audible stimulus at each concentration until the dog awoke. In experiment 2, 12 dogs received the same anesthetic agent they were administered in experiment 1. After duplicate MAC determination, the end tidal concentration was continually decreased by 10% every 15 minutes until the dog awoke from anesthesia (MACawake). RESULTS: Mean induction time was significantly greater for isoflurane-anesthetized dogs (212 seconds), compared with the sevoflurane-anesthetized dogs (154 seconds). Mean+/-SD MACawake-noise was 1.1+/-0.1 vol % for isoflurane and 2.0+/-0.2 vol % for sevoflurane. Mean MAC was 1.3+/-0.2 vol % for isoflurane and 2.1+/-0.6 vol % for sevoflurane, and mean MACawake was 1.0+/-0.1 vol % for isoflurane and 1.3+/-0.3 vol % for sevoflurane. CONCLUSIONS AND CLINICAL RELEVANCE: Sevoflurane resulted in a more rapid induction than did isoflurane. The MACawake for dogs was higher than values reported for both agents in humans. Care should be taken to ensure that dogs are at an appropriate anesthetic depth to prevent consciousness, particularly when single-agent inhalant anesthesia is used. PMID- 18380576 TI - Cyclooxygenase expression and prostanoid production in pyloric and duodenal mucosae in dogs after administration of nonsteroidal anti-inflammatory drugs. AB - OBJECTIVE: To assess cyclooxygenase (COX) expression and prostanoid concentrations in pyloric and duodenal mucosae of dogs after administration of nonsteroidal anti-inflammatory drugs (NSAIDs). ANIMALS: 8 healthy dogs. PROCEDURES: Each dog received carprofen (4.4 mg/kg, q 24 h), deracoxib (2 mg/kg, q 24 h), aspirin (10 mg/kg, q 12 h), and placebo (1 dog treat, q 24 h) orally for 3 days (4-week interval between treatments). Before study commencement (baseline) and on day 3 of each treatment, pyloric and duodenal mucosal appearance was assessed endoscopically and biopsy specimens were obtained for histologic examination. Cyclooxygenase-1 and COX-2 protein expressions were assessed via western blotting, and prostanoid concentrations were measured via ELISAs. An ANOVA was used to analyze data. RESULTS: Treatments had no effect on mucosal appearance and ulceration was not evident histologically. In pyloric and duodenal mucosae, COX-1 expression was unaffected by treatments. Cyclooxygenase-2 expression remained unchanged in pyloric mucosa; in duodenal mucosa, aspirin significantly increased COX-2 expression, compared with effects of deracoxib and carprofen. At baseline, total prostaglandin and thromboxane B2 concentrations in pyloric mucosa were significantly greater than those in duodenal mucosa. Aspirin significantly decreased both prostanoid concentrations in both mucosal tissues, compared with other treatments. In pyloric mucosa, carprofen administration significantly decreased total prostaglandin and thromboxane B2 concentrations, compared with deracoxib administration. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, prostanoid synthesis was greater in pyloric mucosa than it was in duodenal mucosa. Nonselective NSAIDs significantly decreased prostanoid concentrations in these mucosae, compared with the effects of a selective COX-2 NSAID. PMID- 18380577 TI - Relationships between velocities of pulmonary venous flow and plasma concentrations of atrial natriuretic peptide in healthy dogs. AB - OBJECTIVE: To investigate the relationship between velocities of pulmonary venous flow (PVF) and plasma concentrations of atrial natriuretic peptide (ANP) in healthy dogs. ANIMALS: 7 healthy Beagles. PROCEDURES: Dogs were anesthetized, intubated, and positioned in left lateral recumbency. Lactated Ringer's solution was infused (200 mL/kg/h) for 60 minutes via a cephalic vein. Transmitral flow and PVF velocities were measured echocardiographically by use of the apical 4 chamber view. Pulmonary capillary wedge pressure (PCWP) and ANP concentrations were determined. RESULTS: IV infusion significantly increased heart rate and PCWP. Similarly, the ANP concentration significantly increased from baseline (before infusion of lactated Ringer's solution) values. Transmitral flow velocities were significantly increased, although the ratio of velocity of the flow during early ventricular diastole (E wave) to velocity of the atrial flow (A wave; E:A ratio) was unchanged. Regarding the PVF velocities, forward flow during ventricular systole (S wave) and retrograde flow during atrial contraction were significantly increased, whereas velocity of the forward flow during ventricular diastole (D wave) was unchanged. Ratio of the velocity of the S wave to velocity of the D wave was increased significantly, and this ratio was significantly correlated with PCWP or ANP concentration. However, the E:A ratio was not correlated with PCWP or ANP concentration. CONCLUSIONS AND CLINICAL RELEVANCE: PVF velocities were strongly correlated with PCWP and plasma ANP concentration in clinically normal dogs. Therefore, PVF velocities may serve as a sensitive indicator and provide additional information for monitoring acute preloading conditions and estimating atrial filling abnormalities in dogs. PMID- 18380578 TI - Determination of reference intervals for plasma biochemical values in clinically normal adult domestic shorthair cats by use of a dry-slide biochemical analyzer. AB - OBJECTIVE: To establish reference intervals of plasma biochemical values in healthy adult domestic shorthair (DSH) cats by use of controlled conditions. ANIMALS: 95 healthy client-owned cats. PROCEDURES: Food was withheld from the cats overnight. All blood samples were obtained on the same day, at the same location, and by the same investigator. Blood samples were collected from a cephalic vein into lithium heparin tubes. After centrifugation of blood samples, plasma supernatants were harvested and stored at -20 degrees C until assayed for total proteins, albumin, creatinine, urea, glucose, calcium, phosphates, sodium, chloride, potassium, and CO2 concentrations and alkaline phosphatase and alanine aminotransferase activities. RESULTS: Reference intervals in healthy adult DSH cats were 65 to 85 g/L for total proteins, 27 to 39 g/L for albumin, 89 to 207 micromol/L for creatinine, 6.6 to 11.3 mmol/L for urea, 4.1 to 8.2 mmol/L for glucose, 2.4 to 2.9 mmol/L for calcium, 1.1 to 2.1 mmol/L for phosphates, 153 to 161 mmol/L for sodium, 120 to 127 mmol/L for chloride, 3.3 to 4.2 mmol/L for potassium, 15 to 21 mmol/L for CO2, 32 to 147 U/L for alkaline phosphatase, and 34 to 123 U/L for alanine aminotransferase. CONCLUSIONS AND CLINICAL RELEVANCE: This study provided reference intervals for plasma analytes in adult DSH cats. The influence of potential confounding factors was minimized through use of controlled preanalytic and analytic conditions. However, these results cannot be extrapolated to other feline breeds or used to interpret results from other biochemical analyzers. PMID- 18380579 TI - Spectral waveform analysis of intranodal arterial blood flow in abnormally large superficial lymph nodes in dogs. AB - OBJECTIVE: To evaluate pulsed-wave Doppler spectral parameters as a method for distinguishing between neoplastic and inflammatory peripheral lymphadenopathy in dogs. SAMPLE POPULATION: 40 superficial lymph nodes from 33 dogs with peripheral lymphadenopathy. PROCEDURES: 3 Doppler spectral tracings were recorded from each node. Spectral Doppler analysis including assessment of the resistive index, peak systolic velocity-to-end diastolic velocity (S:D) ratio, diastolic notch velocity to-peak systolic velocity (N:S) ratio, and end diastolic velocity-to-diastolic notch velocity ratio was performed for each tracing. Several calculation methods were used to determine the Doppler indices for each lymph node. After the ultrasonographic examination, fine needle aspirates or excisional biopsy specimens of the examined lymph nodes were obtained, and lymphadenopathy was classified as either inflammatory or neoplastic (lymphomatous or metastatic) via cytologic or histologic examination. Results of Doppler analysis were compared with cytologic or histopathologic findings. RESULTS: The Doppler index with the highest diagnostic accuracy was the S:D ratio calculated from the first recorded tracing; a cutoff value of 3.22 yielded sensitivity of 91%, specificity of 100%, and negative predictive value of 89% for detection of neoplasia. Overall diagnostic accuracy was 95%. At a sensitivity of 100%, the most accurate index was the N:S ratio calculated from the first recorded tracing; a cutoff value of 0.45 yielded specificity of 67%, positive predictive value of 81%, and overall diagnostic accuracy of 86.5%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that noninvasive Doppler spectral analysis may be useful in the diagnosis of neoplastic versus inflammatory peripheral lymphadenopathy in dogs. PMID- 18380580 TI - Effects of dietary supplementation with fish oil on in vivo production of inflammatory mediators in clinically normal dogs. AB - OBJECTIVE: To evaluate the effect of diets enriched with eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on in vivo production of interleukin (IL)-1, IL-6, tumor necrosis factor (TNF)-alpha, prostaglandin E2 (PGE2), and platelet activating factor (PAF) in dogs. ANIMALS: 15 young healthy dogs. PROCEDURES: Dogs were randomly allocated to receive an isocaloric ration supplemented with sunflower oil (n=5), fish oil (5), or fish oil plus vitamin E (5) for 12 weeks. At week 12, in vivo production of inflammatory mediators was evaluated in serum at multiple time points for 6 hours following stimulation with IV administration of lipopolysaccharide (LPS). RESULTS: Serum activity or concentration (area under the curve) of IL-1, IL-6, and PGE2 significantly increased after LPS injection in all groups but to a lesser extent in dogs receiving the fish oil diet, compared with results for dogs receiving the sunflower oil diet. Serum activity of TNF alpha and PAF concentration also increased significantly after LPS injection in all groups but did not differ significantly among groups. CONCLUSIONS AND CLINICAL RELEVANCE: A fish oil-enriched diet consisting of 1.75 g of EPA/kg of diet and 2.2 g of DHA/kg of diet (dry-matter basis) with an n-6:n-3 fatty acid ratio of 3.4:1 was associated with significant reductions in serum PGE2 concentrations and IL-1 and IL-6 activities. Results supported the use of EPA- and DHA-enriched diets as part of antiinflammatory treatments for dogs with chronic inflammatory diseases. Additional studies in affected dogs are warranted to further evaluate beneficial anti-inflammatory effects of EPA- and DHA-enriched diets. PMID- 18380581 TI - In vitro evaluation of the effect of trans-10, cis-12 conjugated linoleic acid on phagocytosis by canine peripheral blood polymorphonuclear neutrophilic leukocytes exposed to methylprednisolone sodium succinate. AB - OBJECTIVE: To examine whether in vitro treatment with trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) restores the phagocytic capacity and oxidative burst activity (OBA) of canine polymorphonuclear neutrophilic leukocytes (PMNs) exposed to methylprednisolone sodium succinate (MPSS). SAMPLE POPULATION: Peripheral blood PMNs obtained from 12 healthy Beagles. PROCEDURES: The experimental design involved administration of a high dose of MPSS, which is the recommended protocol for dogs with acute spinal cord injury. To evaluate PMN function, blood samples were collected from dogs before IV injections of doses of MPSS or saline (0.9% NaCl) solution (time 0) and 2, 12, and 24 hours after injections ceased. Polymorphonuclear neutrophilic leukocytes were isolated from blood samples and incubated with t10c12-CLA alone or t10c12-CLA in combination with N acetylcysteine (an antioxidant agent). Phagocytic capacity and OBA were measured simultaneously by use of flow cytometry. RESULTS: The phagocytic capacity and OBA of PMNs were suppressed by IV injection of MPSS and restored 12 hours after injection ceased. In vitro treatment with t10c12-CLA enhanced the phagocytic capacity and OBA of PMNs, regardless of whether dogs had been treated with MPSS. Effects of t10c12-CLA on OBA were detected only when phagocytosis was stimulated by microspheres. Use of N-acetylcysteine attenuated the stimulatory effects of t10c12-CLA. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure to t10c12-CLA enhanced the phagocytic capacity and OBA of canine PMNs, and this effect may have involved t10c12-CLA-induced generation of reactive oxygen species. PMID- 18380582 TI - Effects of inhaled fine dust on lung tissue changes and antibody response induced by spores of opportunistic fungi in goats. AB - OBJECTIVE: To investigate the effects of sterile fine dust aerosol inhalation on antibody responses and lung tissue changes induced by Mucor ramosissimus or Trichoderma viride spores following intratracheal inoculation in goats. ANIMALS: 36 weanling Boer-Spanish goats. PROCEDURES: 6 goats were allocated to each of 2 M ramosissimus-inoculated groups, 2 T viride-inoculated groups, and 2 control (tent or pen) groups. One of each pair of sporetreated groups and the tent control group were exposed 7 times to sterilized fine feedyard dust (mean+/-SD particle diameter, <7.72+/-0.69 microm) for 4 hours in a specially constructed tent. Goats in the 4 fungal treatment groups were inoculated intratracheally 5 times with a fungal spore preparation (30 mL), whereas tent control goats were intratracheally inoculated with physiologic saline (0.9% NaCl) solution (30 mL). Pen control goats were not inoculated or exposed to dust. Goats received an IV challenge with equine RBCs to assess antibody responses to foreign antigens. Postmortem examinations were performed at study completion (day 68) to evaluate lung tissue lesions. RESULTS: 5 of 7 deaths occurred between days 18 and 45 and were attributed to fine dust exposures prior to fungal treatments. Fine dust inhalation induced similar lung lesions and precipitating antibodies among spore treated goats. Following spore inoculations, dust-exposed goats had significantly more spores per gram of consolidated lung tissue than did their nonexposed counterparts. CONCLUSIONS AND CLINICAL RELEVANCE: Fine dust inhalation appeared to decrease the ability of goats to successfully clear fungal spores from the lungs following intratracheal inoculation. PMID- 18380583 TI - Effects of small intestinal ischemia and reperfusion on expression of tumor necrosis factor-alpha and interleukin-6 messenger RNAs in the jejunum, liver, and lungs of dogs. AB - OBJECTIVE: To determine the effects of intestinal ischemia and reperfusion on the expression of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 mRNAs in the jejunum, liver, and lungs of dogs. ANIMALS: 8 healthy adult Beagles. PROCEDURES: In each dog, the cranial mesenteric artery was occluded for 0 (control group; n=4) or 60 (I-R group; 4) minutes, followed by reperfusion for 480 minutes; serum TNF-alpha and IL-6 activities and expression levels of TNF alpha and IL-6 mRNAs in jejunal, hepatic, and lung tissues were measured before and at the end of the ischemic period and at intervals during reperfusion. For each variable, values were compared between the control and I-R groups at each time point. RESULTS: Compared with the control group, serum IL-6 activity increased significantly after 180 minutes of reperfusion in the I-R group; also, jejunal TNF-alpha mRNA expression increased significantly after 60 (peak) and 180 minutes of reperfusion. In the I-R group, expressions of IL-6 mRNA in the liver and TNF-alpha and IL-6 mRNAs in the lungs increased significantly at 480 minutes of reperfusion, compared with the control group. Serum TNF-alpha activity, expression of IL-6 mRNA in the jejunum, and expression of TNF-alpha mRNA in the liver in the control and I-R groups did not differ. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the liver, lungs, and jejunum contributed to the production of TNF-alpha and IL-6 after intestinal ischemia and reperfusion in dogs, suggesting that intestinal ischemia and reperfusion induce a systemic proinflammatory cytokine response in dogs. PMID- 18380585 TI - Effect of body position on intraocular pressure in dogs without glaucoma. AB - OBJECTIVE: To determine the effects of body position on intraocular pressure (IOP) in dogs without glaucoma. ANIMALS: 24 healthy dogs with no evidence of glaucoma. PROCEDURES: Dogs underwent ophthalmic examinations to ensure that no IOP-affecting ocular diseases were present. Each dog was sequentially placed in dorsal recumbency, sternal recumbency, and sitting position. For each of the 3 positions, IOP in the right eye was measured by use of an applanation tonometer immediately after positioning (0 minutes) and after 3 and 5 minutes had elapsed. The initial body position was randomly assigned; each position followed the other positions an equal number of times, and IOP measurements were initiated immediately after moving from one body position to the next. Proparacaine hydrochloride (0.5%) was applied to the right eye immediately prior to IOP measurements. RESULTS: Intraocular pressure was affected by body position. During the 5-minute examination, IOP decreased significantly in dogs that were dorsally recumbent or sitting but did not change significantly in dogs that were sternally recumbent. For the 3 positions, overall mean IOP differed significantly at each time point (0, 3, and 5 minutes). Mean IOP in dorsal recumbency was significantly higher than that in sternal recumbency at 0 and at 3 minutes; although the former was also higher than that in sitting position at 3 minutes, that difference was not significant. CONCLUSIONS AND CLINICAL RELEVANCE: Body position affects IOP in dogs. When IOP is measured in dogs, body position should be recorded and consistent among repeat evaluations. PMID- 18380584 TI - Biological activity of dihydroartemisinin in canine osteosarcoma cell lines. AB - OBJECTIVE: To evaluate the biological activity of dihydroartemisinin on canine osteosarcoma cell lines in vitro. SAMPLE POPULATION: 4 canine osteosarcoma cell lines. PROCEDURES: Cell viability assays were performed on canine osteosarcoma cell lines OSCA2, OSCA16, OSCA50, and D17 after 24, 48, and 72 hours of treatment with dihydroartemisinin at concentrations of 0.1 to 100 microM. Apoptosis was assessed by use of an ELISA for free nuclosomal DNA fragmentation and by western blot analysis for cleavage of caspase 3. Cell cycle analysis was performed by use of staining with propidium iodide and flow cytometry. Detection of reactive oxygen species (ROS) was conducted in the D17 cell line by use of 6-carboxy-2',7' dihydrofluorescein diacetate and flow cytometry. RESULTS: The concentration of dihydroartemisinin required for 50% inhibition of cell viability (IC50) was achieved in all 4 canine osteosarcoma cell lines and ranged from 8.7 to 43.6 microM. Induction of apoptosis was evident as an increase in nucleosomal DNA fragmentation, cleavage of caspase 3, and an increase in the population in the sub G0/G1 phase of the cell cycle detected by flow cytometry. Exposure to dihydroartemisinin also resulted in a decrease in the G0/G1 population. Iron dependent generation of ROS was detected in dihydroartemisinin-treated D17 cells; ROS generation increased in a dose-dependent manner. CONCLUSIONS AND CLINICAL RELEVANCE: Incubation with dihydroartemisinin resulted in biological activity against canine osteosarcoma cell lines, which included induction of apoptosis and arrest of the cell cycle. Clinical trials of dihydroartemisinin in dogs with osteosarcoma should be conducted. PMID- 18380586 TI - Pharmacokinetics of remifentanil in conscious cats and cats anesthetized with isoflurane. AB - OBJECTIVE: To characterize the pharmacokinetics of remifentanil in conscious cats and cats anesthetized with isoflurane. ANIMALS: 6 cats. PROCEDURES: Remifentanil (1 microg/kg/min for 5 minutes) was administered IV in conscious cats or cats anesthetized with 1.63% isoflurane in oxygen in a randomized crossover design. Blood samples were obtained immediately prior to remifentanil administration and every minute for 10 minutes, every 2 minutes for 10 minutes, and every 5 minutes for 10 minutes after the beginning of the infusion. Blood was immediately transferred to tubes containing citric acid, flash frozen in liquid nitrogen, and stored at -80 degrees C until analysis. Blood remifentanil concentration was determined by use of liquid chromatography-mass spectrometry. Remifentanil concentration-time data were fitted to compartment models. RESULTS: A 2 compartment model (with zero-order input because of study design) best described the disposition of remifentanil in awake and isoflurane-anesthetized cats. The apparent volume of distribution of the central compartment, the apparent volume of distribution at steady state, the clearance, and the terminal half-life (median [range]) were 1,596 (1,164 to 2,111) and 567 (278 to 641) mL/kg, 7,632 (2,284 to 76,039) and 1,651 (446 to 29,229) mL/kg, 766 (408 to 1,473) and 371 (197 to 472) mL/min/kg, and 17.4 (5.5 to 920.3) and 15.7 (3.8 to 410.3) minutes in conscious and anesthetized cats, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The disposition of remifentanil in cats was characterized by a high clearance. Isoflurane anesthesia significantly decreased the volume of the central compartment, likely by decreasing blood flow to vessel-rich organs. PMID- 18380587 TI - Evaluation of an endoscopically assisted gastropexy technique in dogs. AB - OBJECTIVE: To evaluate the use of endoscopy in conjunction with a gastropexy technique in dogs as a potential means to aid prevention of gastric dilatation volvulus. ANIMALS: 12 healthy adult medium- and large-breed dogs. PROCEDURES: 12 adult research dogs that had no abnormal physical examination findings each underwent an endoscopically assisted gastropexy procedure. On completion of the procedure, the dogs were euthanized and exploratory laparotomies were performed to evaluate the surgical site. Data recorded included anatomic location of the gastropexy, gastropexy length, and duration of procedure as well as any complications. RESULTS: Mean+/-SD gastropexy length was 3.3+/-0.25 cm, and mean duration of surgery was 18+/-7 minutes. In each dog, the stomach was located in its normal anatomic position and all gastropexies were sutured to the abdominal wall at the level of the pyloric antrum. The only complications during the procedure were needle bending and breakage at the time of stay suture placement. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of these findings, it appears that endoscopically assisted gastropexy is a simple, fast, safe, and reliable method of performing a prophylactic gastropexy in dogs when undertaken by a person who is skilled in endoscopy. Such a procedure maximizes the benefits of decreased morbidity and shorter duration of anesthesia associated with minimally invasive surgery. Further clinical studies are warranted to evaluate the long term efficacy of this procedure in dogs at risk for development of gastric dilatation-volvulus. PMID- 18380588 TI - Ventral abdominal approach for screw fixation of sacroiliac luxation in cadavers of cats and dogs. AB - OBJECTIVE: To investigate a technique for repair of sacroiliac luxation with positional screw insertion from the ventral surface of the sacral wing via a ventral abdominal approach. SAMPLE POPULATION: Hemipelvis specimens from cadavers of 5 small- to large-breed dogs and 9 European shorthair cats. PROCEDURES: An optimal entry point and a safe drill corridor for implant placement were determined (4 hemipelvis specimens). Anatomic landmarks were identified, and the surgical technique for a ventral abdominal approach was described. Single positional screw placement was performed across the sacroiliac joint in 23 hemipelvis specimens. Screws were aimed at 25 degrees (n=2), 35 degrees (2), and 45 degrees (19) angles to the vertical axis in a transverse plane (alpha angles) and at a 90 degrees angle to the longitudinal axis in a dorsal plane (beta angle). Implant placement was assessed by radiographic evaluation of the cadavers and of the hemipelvis specimens devoid of soft tissue. RESULTS: By use of alpha angles of 35 degrees and 45 degrees, 20 of 21 implants were placed adequately; screws crossed the sacroiliac joint and penetrated the wing of the ilium without damaging adjacent nerves. The measured median alpha angle was 38 degrees , and the median beta angle was 88 degrees. One complication was recorded. CONCLUSIONS AND CLINICAL RELEVANCE: Cortical positional screw placement from the ventral aspect of the sacral wing by use of a ventral abdominal approach could be an alternative to conventional techniques. This novel technique may be useful for repair of bilateral sacroiliac luxation, treatment of concomitant soft tissue injuries of the caudal portion of the abdominal cavity or abdominal wall, and repair of pelvic floor fractures in a single approach. PMID- 18380589 TI - Ventral abdominal approach for screw fixation of sacroiliac luxation in clinically affected cats. AB - OBJECTIVE: To investigate sacroiliac luxation repair with positional screw insertion from the ventral surface of the sacral wing via a ventral abdominal approach in cats. ANIMALS: 18 European shorthair cats. PROCEDURES: All cats underwent clinical examination including orthopedic and neurologic examination and assessment of lameness and pain before and immediately after surgery and 6 and 16 weeks after surgery. All sacroiliac luxations were stabilized with a single positional 2.4-mm cortical titanium self-tapping screw. The pelvic floor was also repaired in selected cats. Screw entry points and angles determined in a prior study of cadavers were used. Radiographs were taken before surgery and during follow-up evaluations to assess postoperative sacroiliac luxation reduction, implant placement, and repair stability. RESULTS: All implants were placed correctly. Iatrogenic sciatic nerve injuries occurred in 2 cats. Median time to ambulation was 1.5 days for cats with sacroiliac luxation as the sole injury. Radiographic outcome of sacroiliac luxation repair was excellent in 15 of 17 repairs, good in 1 of 17 repairs, and poor in 1 of 17 repairs. Clinical outcome was excellent in 11 of 15 cats and good in 4 of 15 cats. CONCLUSIONS AND CLINICAL RELEVANCE: Insertion of a positional screw across the sacroiliac joint via a ventral abdominal approached can be an alternative to conventional techniques of sacroiliac luxation repair in cats. This novel technique allowed repair of bilateral sacroiliac luxation, repair of pelvic floor fractures, and treatment of soft tissue injuries of the abdominal cavity or abdominal organs with a single approach. PMID- 18380590 TI - Investigation of the role of healthy dogs as potential carriers of rabies virus. AB - To investigate whether healthy animals are potential carriers of rabies virus in China, 153 domestic dogs were collected from a rabies enzootic area, Anlong county in Guizhou Province, and monitored for 6 months. Initially, findings of rabies virus antigen in the saliva of 15 dogs by an enzyme-linked immunosorbent assay (ELISA) test suggested they might be carriers. These 15 dogs were kept under observation for 6 months. None of the dogs showed any clinical signs of rabies during the observation period. Moreover, using the ELISA test alone, detection of rabies virus antigen in saliva of some animals was not consistent during the observation period. However, none of the saliva samples collected either at the time of acquisition or during the observation period was found to be positive for rabies virus RNA by reverse transcriptase-polymerase chain reaction (RT-PCR). Furthermore, neither viral antigen nor viral RNA was detected in the brain samples collected at the time of euthanasia. These results do not provide support for the contention that healthy dogs act as carriers in rabies. Caution is urged when preliminary and nondefinitive tests, such as ELISA, are used to infer clinical status related to rabies. PMID- 18380591 TI - Diagnostics of Pogosta disease: antigenic properties and evaluation of Sindbis virus IgM and IgG enzyme immunoassays. AB - Sindbis virus (SINV) is a mosquito-borne causative agent of a fever-rash arthritis, Pogosta disease, as verified recently by virus isolation from acutely ill patients. Pogosta disease occurs annually, but it emerges as unique epidemics every 7 years in Finland; over 10,000 patient samples have been analyzed for SINV antibodies, with over 2000 diagnosed acute SINV infections. However, the performance of these serological tests with a large number of samples has not been described before. The aim of the present study was to characterize and evaluate methods developed for the serodiagnostics of SINV infection, suitable for large sample numbers, and to examine the protein-specific responses to the antigen used. We developed SINV IgM and IgG enzyme immunoassays (EIA) using highly purified SINV. The EIAs were compared to hemagglutination inhibition (HI) and neutralization tests. We studied paired samples from 46 acutely ill patients taken at approximately 2-week intervals, with a verified SINV infection confirmed by a fourfold rise in HI antibody titer. The assay cut-off values and specificity were determined with confirmed negative sera. Protein-specific antibody responses were examined with immunoblot assay. The optical density values of the EIAs correlated well with the HI titers. The sensitivities of the IgM and IgG EIAs were 97.6% and 100%, and specificities were 95.2% and 97.6%, respectively. We consider that a verified serological diagnosis of acute SINV infection requires (1) in addition to a positive IgM result at least a fourfold increase in HI (or IgG) titer between paired sera or (2) a positive IgM result and a negative/borderline IgG result (which excludes old immunity) and specific reaction in HI. Both E1 and E2 glycoproteins of SINV were shown to be recognized by IgM and IgG antibodies early in infection. PMID- 18380592 TI - Sequential biochemical and mechanical stimulation in the development of tissue engineered ligaments. AB - Application of stimuli in sequence to developing cultures in vitro offers the potential to intricately direct cell development and differentiation by following the template of native tissue behavior. We hypothesize that administration of mechanical stimulation at the peak of growth factor-induced cell activity will differentiate bone marrow stromal cells (BMSCs) along a fibroblast lineage and enhance in vitro ligament development through enhanced matrix ingrowth, matrix metalloproteinase-2 (MMP-2) production, collagen type I production, and extracellular matrix (ECM) alignment. BMSC-seeded silk matrices were cultured in a static growth-factor-free environment for 5 days prior to loading into bioreactor vessels to first establish an appropriate dynamic rotational regime, as determined through assessment of cell activity, histology, and surface topography. Once the regime was determined, seeded matrices initially cultured in basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), or growth factor-free control medium for 5 days were loaded into the bioreactor for 9 days of mechanical stimulation. Our findings indicated that the sequential application of mechanical stimulation following growth factor supplemented static culture induced cell differentiation toward a fibroblast lineage, enhancing matrix ingrowth, cell and ECM alignment, and total collagen type I produced compared to respective static cultures. The current results suggest a dynamic culturing regime in the development of engineered tissues. PMID- 18380593 TI - Bombyx mori silk fibroin membranes as potential substrata for epithelial constructs used in the management of ocular surface disorders. AB - Membranes were prepared from fibroin, a protein isolated from the domesticated silkworm (Bombyx mori) silk, and evaluated as a potential substratum for corneal limbal epithelial cells. These membranes (i.e., B. mori silk fibroin [BMSF] membranes) were cast from dialyzed solutions of fibroin protein (4% w/v) dispensed into 35-mm-diameter culture dishes and dried at room temperature (23-24 degrees C). The resulting material was transparent, easy to handle, and supported levels of human limbal epithelial (HLE) cell growth comparable to that observed on tissue culture plastic. Remarkably, these results were obtained utilizing a commercial serum-free medium (CnT-20) designed for the ex vivo expansion of corneal epithelial progenitor cells. The potential benefits of serum proteins on this culture system were examined through addition of fetal bovine serum (FBS) either to fibroin stocks prior to membrane casting or by supplementation of the CnT-20 medium. Membranes cast in the presence of FBS displayed increasing opacity and induced little change in HLE growth. Supplementation of CnT-20 medium with FBS deterred cell growth on all substrata, including tissue culture plastic control substrata. The remarkable properties of BMSF membranes demonstrated under serum-free conditions warrant investigation of this material as a substratum in the creation of tissue-engineered constructs for the restoration of diseased or damaged ocular surface. PMID- 18380594 TI - Mucosal biofilms: challenges and future directions. PMID- 18380595 TI - Foreigners in our body. PMID- 18380597 TI - Micafungin for the prophylaxis and treatment of Candida infections. AB - Invasive fungal infection is a significant cause of morbidity and mortality worldwide. The incidence of these infections is steadily increasing. In addition, strains resistant to many commonly used antifungal agents are becoming more prevalent. Many new antifungals have become commercially available in recent years, which have vastly improved the ability to treat these infections effectively. Micafungin is one of three commercially available echinocandins available for use in the USA. This class of agents possess a unique mechanism of action that helps to reduce toxicity while maintaining potent antifungal activity. Micafungin is currently approved for the treatment of esophageal candidiasis in adults and is the only in its class approved for the prophylaxis of Candida infection in patients who have undergone hematopoietic stem cell transplantation. It was also recently approved in the USA for the treatment of candidemia and other forms of invasive candiaisis (acute disseminated candiaisis, Candida peritonitis and abscess). In general, micafungin is well tolerated and has favorable safety and drug-interaction profiles. PMID- 18380598 TI - Opportunistic pathogens in AIDS: trends, diagnosis and priorities. AB - The second International Conference on Opportunistic Pathogens in AIDS was organized by the Department of Laboratory Medicine, All India Institute of Medical Sciences (New Delhi, India). The conference was held at the All India Institute of Medical Sciences in India between the 27th and 29th January 2008, and was attended by 225 participants representing 21 countries. The conference focused on trends, diagnosis and management of coinfections among HIV patients. In addition, as part of this conference, a symposium was held on the links between substance abuse, HIV and neuropathology. PMID- 18380599 TI - Pediatric HIV infection: the state of antiretroviral therapy. AB - Pediatric HIV/AIDS has become less of a problem in resource-rich countries as the number of perinatal infections has reduced dramatically since the advent of antiretrovirals, resulting in the effective prevention of mother-to-child transmission. In resource-limited settings, however, pediatric HIV infection remains a colossal problem; a separate review in this same issue of Expert Review of Anti-Infective Therapy examines the international aspects of pediatric HIV/AIDS. Treatment of HIV infection in children differs from that in adults in the use of immunologic markers and owing to drug pharmacokinetics and age-related adherence issues. This review, geared for the general pediatrician or family practitioner who may see the HIV-positive child in the clinic or the hospital, summarizes the most recent pediatric data and guidelines for the testing and treatment of HIV, including the US NIH guidelines released in February 2008. Treatment-experienced patients, who should be cared for by pediatric HIV specialists, are not addressed here specifically. Adolescents, infected either perinatally or sexually, with their own unique issues, deserve a separate review. PMID- 18380600 TI - HIV care and treatment for children in resource-limited settings. AB - Although efforts to combat the HIV epidemic have focused on the perinatal reduction of HIV transmission, many children are still being infected with HIV in resource-limited settings. Access to HIV care, cotrimoxazole and antiretroviral therapy (ART) for HIV-infected children has greatly improved in recent years, and has proven to be very effective in reducing mortality in all age categories. Many challenges remain to be resolved, such as the retention in care of children born to HIV-infected mothers, the lack of pharmacokinetic data on ART in malnourished children, optimum timing of ART, treatment and diagnosis of concomitant tuberculosis, and the effects of ART and HIV on the child's development. In the long term, treatment success might be negated due to lower rates of viral suppression in children and the accumulation of resistance mutations. Evidenced based comprehensive care models should allow for decentralizing care up to the level of the community, allowing larger numbers of children to receive HIV care. PMID- 18380601 TI - Treatment of chronic hepatitis B and the implications of viral resistance to therapy. AB - Treatment options for chronic HBV infection have improved considerably in recent years. However, with the increased number of available antiviral drugs, it is also more difficult to select the optimal drug for each individual patient. Antiviral potency, the risk for the development of drug resistance, safety and costs of treatment all need to be considered. In this review, we describe current concepts in the treatment of chronic hepatitis B, with a special focus on the importance of drug resistance and its implications for the long-term management of hepatitis B. Suggestions for avoiding resistance and management of established drug resistance are provided for each individual approved compound. PMID- 18380603 TI - Antimicrobial stewardship programs: interventions and associated outcomes. AB - Guidelines regarding antimicrobial stewardship programs recommend an infectious diseases-trained physician and an infectious diseases-trained pharmacist as core members. Inclusion of clinical microbiologists, infection-control practitioners, information systems experts and hospital epidemiologists is considered optimal. Recommended stewardship interventions include prospective audit and intervention, formulary restriction, education, guideline development, clinical pathway development, antimicrobial order forms and the de-escalation of therapy. The primary outcome associated with these interventions has been the associated cost savings; however, few published investigations have taken into account the overall cost of the intervention. Over the past 5 years, there has been an increased focus upon interventions intended to decrease bacterial resistance or reduce superinfection, including infections associated with Clostridium difficile colitis. Few programs have been associated with a reduction in antimicrobial drug adverse events. Antimicrobial stewardship programs are becoming increasingly associated with clear benefits and will be integral in the in-patient healthcare setting. PMID- 18380602 TI - Pathogenesis of mucosal biofilm infections: challenges and progress. AB - Living-tissue biofilms remained unrecognized until very recently, mainly as a result of traditional microbial sampling techniques or histologic processing, which disrupt the spatial organization of the tissue microorganisms. Thus, the biofilm nature of certain mucosal infections was frequently unintentionally missed or disregarded. To a large extent, the study of human tissue biofilms is still in its infancy. However, with the advent of newer methodologies, such as fluorescent in situ hybridization and endoscopic confocal laser scanning microscopy, which combine the identification of microbes with in situ, direct visualization of their relationships with each other and with their substratum, mucosal tissue biofilms are becoming easier to study and, thus, their role in human infections is becoming more apparent. This review summarizes the challenges in the study of tissue biofilms, proposes two inflammation-centered - albeit opposite - pathogenetic models of mucosal tissue biofilm infections and suggests directions for future research and novel therapeutic approaches. PMID- 18380604 TI - Acquired rifamycin resistance: pharmacology and biology. AB - The global emergence of multidrug-resistant and extensively drug-resistant TB has refocused attention on preventing acquired resistance. This article reviews basic science, pharmacology and public policy to understand the contribution of these factors to acquired rifamycin resistance (ARR), a critical factor in multidrug resistance. Directly observed therapy short course (DOTS) effectiveness requires that each drug's effects persist equally throughout the dosing interval. Although rifampin and isoniazid have similar pharmacokinetics, the postantibiotic effect of rifampin is several times greater than that of isoniazid. As a result, rifampin's effects may persist unopposed when standard multidrug regimens are administered at intervals longer than 24 h. ARR may occur in this setting when outgrowth is not otherwise contained by the host immune response. Most countries do not provide weekend therapy under DOTS. Limiting TB therapy to weekdays may promote the emergence of acquired drug resistance in patients with advanced AIDS and TB. A large, simple trial to examine this question is both warranted and feasible. PMID- 18380605 TI - Tularemia: current diagnosis and treatment options. AB - Tularemia is an infection caused by Francisella tularensis with a worldwide distribution and diverse clinical manifestations. Limitations in both culture and serologic testing have led to substantial research into new diagnostic techniques and their clinical application, with PCR testing as the best example. This review focuses on the utility of culture, PCR and serologic testing for tularemia. In addition, we also review the evidence to support different therapeutic options for tularemia, highlighting both the most effective supporting evidence for therapeutic recommendations as well as gaps in current knowledge. We conclude the article with suggestions regarding potential areas for future research. PMID- 18380606 TI - Management of Lyme disease. AB - It has been 30 years since Lyme disease was first described in a cohort of patients from Connecticut. An understanding of disease transmission, clinical manifestations and prevention strategies has been established. With the number of new cases increasing each year, it is important that clinicians are aware of the available treatment options. Most patients respond well to a course of treatment with a recommended antibiotic; however, for those patients who develop post-Lyme disease syndrome, the management is unclear. This review provides an overview of Lyme disease and the recommended treatment options available to physicians. PMID- 18380607 TI - Treatment options for paracoccidioidomycosis and new strategies investigated. AB - Paracoccidioidomycosis is the most prevalent systemic endemic mycosis in South America with most reported cases in Brazil. It is a major cause of disability and death among young adult rural workers during their most productive years of life. Sequels are frequent and the evolution of the disease and mortality burden are strongly influenced by the socio-economic status of the patients. Although long periods of antifungal therapy (sulfamethoxazole/trimethoprim, itraconazole and amphotericin B) are used in clinical practice, relapses remain a significant unresolved problem. Early diagnosis is hampered by structural factors, ranging from the high costs of reagents, the lack of trained personnel and limited access to the healthcare system by rural workers. A peptide vaccine aimed at immunotherapy of paracoccidioidomycosis, as an adjuvant to chemotherapy, is being studied. The protective effects obtained in mice intratracheally infected with Paracoccidioides brasiliensis, and the promiscuous binding of the peptide P10 to HLA-DR molecules, suggest that it could be used as a vaccine to reduce the duration of chemotherapy and the risk of relapse. PMID- 18380609 TI - Use of complementary and alternative medicine by physicians in St. Petersburg, Russia. AB - BACKGROUND: Interest in complementary and alternative medicine (CAM) is increasing worldwide, although relatively little is known of physician use of CAM, and there are no quantitative reports of CAM use by Russian physicians. OBJECTIVE: The objective of this study was to determine the prevalence of CAM therapies among practicing physicians and determine predictors of CAM usage. DESIGN: This was a convenience sample prevalence survey. SETTING: The study involved 3 urban academic hospitals in St. Petersburg, in Russia. SUBJECTS: Participants included 192 physicians practicing at the three study hospitals. MEASUREMENTS: The study determined the number (from a list of 32) of CAM therapies that physicians used on themselves, used on their patients, or referred their patients to receive. RESULTS: One hundred and seventy-seven (177; 92%) of the surveyed physicians responded. One hundred percent (100%) of the respondents had practiced CAM or referred patients for at least two CAM therapies. One hundred and seventy-five (175; 99%) had themselves practiced at least two therapies. On average, each physician had practiced or referred patients for 12.7 (95% confidence interval, 11.9-13.6) therapies. On multivariate analysis, knowledge of a foreign language, surgical specialty, and female gender were significantly (p < 0.05) associated with increased CAM usage, while critical care specialty and completion of only an internship were associated with lower rates of CAM practice. CONCLUSIONS: Physician use of CAM in Russia appears very high. The high prevalence of CAM may complicate adoption of Western evidence-based practices. Predictors and effects of CAM usage in Russian society warrant further study. PMID- 18380610 TI - More capital punishment and the veterinary profession. PMID- 18380611 TI - A new understanding for handling disaster. PMID- 18380612 TI - Offering solutions to pet obesity. PMID- 18380613 TI - What is your diagnosis? Noncardiogenic pulmonary edema secondary to upper airway obstruction. PMID- 18380614 TI - Veterinary practice expenses and financial ratios, 2005. PMID- 18380615 TI - Postmarketing surveillance of rabies vaccines for dogs to evaluate safety and efficacy. PMID- 18380616 TI - Serum antibodies against human albumin in critically ill and healthy dogs. AB - OBJECTIVE: To characterize the magnitude and duration of the antibody response against human albumin (HA) in critically ill and healthy dogs. DESIGN: Cohort and cross-sectional study. ANIMALS: Fourteen critically ill dogs that received 25% HA as part of their treatment protocol, 2 healthy dogs with no known previous exposure to HA that received 2 infusions of 25% HA (positive control dogs), and 47 healthy dogs and 21 critically ill dogs with no known exposure to HA (negative control dogs). PROCEDURES: An ELISA to detect IgG against HA was developed. Serum samples were obtained from the critically ill dogs prior to infusion of HA, at the time of hospital discharge, and 4 to 6 weeks and 6 months after HA administration. Serum samples were obtained at 2- to 4-week intervals from both positive control dogs for 101 weeks. A single serum sample was obtained from each of the negative control dogs. RESULTS: All 14 critically ill dogs developed serum IgG against HA. Peak antibody response was detected 4 to 6 weeks after HA administration. In both positive control dogs, IgG against HA was detected 10 days after HA administration and continued past 97 weeks. The peak antibody response was detected at 3 weeks in 1 dog and at 9 weeks in the other. Five of the 68 (7%) negative control dogs had a positive antibody response. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that dogs developed a pronounced IgG response following exposure to HA and that some dogs with no history of HA administration were positive for anti-HA IgG. PMID- 18380617 TI - Pyogranulomatous cystitis associated with Toxoplasma gondii infection in a cat after renal transplantation. AB - CASE DESCRIPTION: An 8-year-old spayed female domestic shorthair cat was evaluated for azotemia and a suspected mass in the urinary bladder 6 weeks after receiving a renal transplant. Ultrasonography revealed a mass at the ureteroneocystostomy site, and the mass was excised. Both the donor and recipient cats were seronegative for Toxoplasma gondii-specific IgG antibodies prior to transplantation. CLINICAL FINDINGS: Histologic evaluation of the mass revealed lesions indicative of extensive necrotizing pyogranulomatous cystitis with numerous intralesional T gondii tachyzoites and bradyzoite cysts. TREATMENT AND OUTCOME: Treatment with clindamycin was initiated; however, the cat's clinical condition continued to decline, and it was euthanized 9 days after the mass was excised. Necropsy revealed T gondii cysts within the renal allograft and the transplanted ureter, with no evidence of systemic spread of organisms. CLINICAL RELEVANCE: Toxoplasmosis should be considered as a differential diagnosis for azotemia in feline renal transplant recipients regardless of the results of assays for T gondii antibodies in the serum of donors or recipients. This report illustrated the need for improved screening of donor and recipient cats and the importance of minimizing exposure to potential sources of T gondii after transplantation. PMID- 18380618 TI - Preputial damage and lateral penile displacement during castration in a degu. AB - CASE DESCRIPTION: A 2-month-old male degu was treated for preputial damage and lateral penile displacement that occurred during attempted castration. CLINICAL FINDINGS: Bruising and swelling of the prepuce and severe edema to the left of the prepuce were evident. The penis could not be extruded from the prepuce. Radiography revealed a large bladder. Contrast medium injected into the prepuce filled the peripreputial subcutaneous tissues. TREATMENT AND OUTCOME: During surgical exploration through a peripreputial approach, the penis was found to be completely separated from the prepuce and located in the adjacent subcutaneous tissue. The penis was repositioned in the prepuce and anchored with a suture at its base. The following day, the preputial orifice was crusted over, urine was leaking from the incision, and the penis could not be extruded from the prepuce. The anchoring suture was removed, and the tip of the penis was sutured to the preputial orifice so that the penis protruded slightly from the prepuce. Urination was normal after the second surgery. Two years later, the preputial orifice remained adhered to the distal portion of the penis and the exposed penile tissue was healthy. CLINICAL RELEVANCE: Penile displacement from the prepuce is an unusual complication of castration in degus. The surgical technique used in this animal may be an effective means of repair. Permanent exposure of the tip of the penis may be well-tolerated in degus. PMID- 18380619 TI - Bilateral pubic and ischial osteotomy for surgical management of caudal colonic and rectal masses in six dogs and a cat. AB - CASE DESCRIPTION: 6 dogs and a cat were evaluated because of caudal colonic and rectal masses. CLINICAL FINDINGS: Tumors were identified in the caudal portion of the colon (n = 2), in the area of the colorectal junction (2), or in the rectum (3). TREATMENT AND OUTCOME: In all 7 animals, bilateral pubic and ischial osteotomy was performed to provide exposure of the rectum and associated tumor. Masses were successfully removed, and all 7 animals were able to ambulate normally within 3 days after surgery. No complications associated with the osteotomy procedure were identified. CLINICAL RELEVANCE: Findings suggested that bilateral pubic and ischial osteotomy provided sufficient exposure for resection of intrapelvic tumors in dogs and a cat with minimal complications. PMID- 18380620 TI - Esophageal foreign body obstruction caused by a dental chew treat in 31 dogs (2000-2006). AB - OBJECTIVE: To evaluate the signalment, clinical signs, diagnosis, treatment, and outcome associated with esophageal obstruction caused by a dental chew treat in dogs. DESIGN: Retrospective case series. ANIMALS: 31 dogs. PROCEDURES: Medical records were contributed from 19 hospitals via responses to a questionnaire that was developed for veterinarians who managed the cases. RESULTS: Esophageal obstructions with the dental chew treat occurred primarily in small dogs (26/31 [83.9%]). The most common clinical signs were gagging, regurgitation, vomiting, anorexia, and lethargy. Diagnosis was usually made via survey thoracic radiography. Obstructions were most commonly located in the distal portion of the esophagus (23/31; 74.2%). Esophageal lesions were moderate or severe in 86.7% (26/30) of the dogs. Orad endoscopic removal of the foreign bodies was uncommon (8/31 [25.8%]); most were pushed into the stomach. Thoracotomy was necessary in 6 dogs. Esophageal strictures developed in 6 of 25 (24%) dogs that survived initial hospitalization. Overall mortality rate was 25.8%. CONCLUSIONS AND CLINICAL RELEVANCE: Esophageal obstructions with a dental chew treat were difficult to remove orally via endoscopy, resulted in moderate or severe esophageal damage, frequently were associated with stricture formation, and were associated with a high mortality rate. PMID- 18380622 TI - Use of cystoscopic-guided laser ablation for treatment of intramural ureteral ectopia in male dogs: four cases (2006-2007). AB - OBJECTIVE: To describe and evaluate the outcome of cystoscopic-guided laser ablation of intramural ureteral ectopia in male dogs. DESIGN: Retrospective case series. ANIMALS: 4 incontinent male dogs with intramural ureteral ectopia. PROCEDURES: Intramural ectopic ureters were diagnosed via preoperative computed tomography-IV urography and subsequent cystoscopy. Transurethral cystoscopic guided laser ablation (diode laser [n = 3 dogs] and holmium:yttrium aluminum garnet laser [1]) was performed to proximally relocate the ectopic ureteral orifice to the urinary bladder. Fluoroscopy was used during the procedures to confirm that the ureteral tract was intramural and the ureteral orifice was intravesicular after the procedure. In 1 dog with bilateral ureteral ectopia, staged laser ablation was performed at 6-week intervals because of difficulty viewing the second ureter on the first attempt. All ureteral orifices were initially located in the middle to proximal portion of the prostatic portion of the urethra. Six weeks after surgery, imaging was repeated in 3 of 4 dogs. RESULTS: Postoperative dysuria or hematuria did not develop. All dogs were immediately continent after laser treatment and remained so at a median follow-up period of 18 months (range, 15 to 20 months) without medical management. CONCLUSIONS AND CLINICAL RELEVANCE: Ureteral ectopia can cause urinary incontinence in male dogs and is usually associated with other urinary tract abnormalities. Cystoscopic-guided laser ablation provided an effective and minimally invasive alternative to surgical management of intramural ureteral ectopia. PMID- 18380623 TI - Chemoprophylactic effects of azithromycin against Rhodococcus equi-induced pneumonia among foals at equine breeding farms with endemic infections. AB - OBJECTIVE: To determine the effect of azithromycin chemoprophylaxis on the cumulative incidence of pneumonia caused by Rhodococcus equi, age at onset of pneumonia, and minimum inhibitory concentration (MIC) of azithromycin for R equi isolates cultured from fecal and clinical samples. DESIGN: Controlled, randomized clinical trial. ANIMALS: 338 foals born and raised at 10 equine breeding farms; each farm had a history of endemic R equi infections. PROCEDURES: Group 1 foals were control foals, and group 2 foals were treated with azithromycin (10 mg/kg [4.5 mg/lb], PO, q 48 h) during the first 2 weeks after birth. Foals were monitored for development of pneumonia attributable to R equi infection and for adverse effects of azithromycin. Isolates of R equi were tested for susceptibility to azithromycin. RESULTS: The proportion of R equi-affected foals was significantly higher for control foals (20.8%) than for azithromycin-treated foals (5.3%). Adverse effects of azithromycin treatment were not detected, and there were no significant differences between groups for the MICs of azithromycin for R equi isolates cultured from fecal or clinical samples. CONCLUSIONS AND CLINICAL RELEVANCE: Azithromycin chemoprophylaxis effectively reduced the cumulative incidence of pneumonia attributable to R equi among foals at breeding farms with endemic R equi infections. There was no evidence of resistance to azithromycin. Nonetheless, caution must be used because it is possible that resistance could develop with widespread use of azithromycin as a preventative treatment. Further investigation is needed before azithromycin chemoprophylaxis can be recommended for control of R equi infections. PMID- 18380624 TI - Enhancement of plasmid DNA yields during fed-batch culture of a fruR-knockout Escherichia coli strain. AB - Well-characterized derivatives of Escherichia coli K12 such as DH5alpha are the host strains commonly used for plasmid DNA production. Owing to the prospective clinical demand for large quantities of plasmid DNA for gene therapy and DNA vaccination, existing plasmid production processes need to be optimized to attain higher plasmid yields. To date, nearly all production optimization efforts are focused on media or fermentation process design. Although there has been a simple empirical evaluation of the available host strains, there is a lack of systematic effort at engineering these host strains for improved plasmid DNA production. In view of this, we engineered DH5alpha WT (wild-type) cells carrying a DNA vaccine plasmid by knocking out the fruR (fructose repressor) [also known as the Cra (catabolite repressor activator)] global regulator gene and evaluated the growth and plasmid yields of these P+ (plasmid-bearing) fruR cells (fruR-knockout cells) during fed-batch cultures with exponential feeding. The P+ fruR cells showed a more rapid accumulation of plasmid DNA towards the end of the fed-batch cultures compared with the P+ WT cells. As a result, the specific plasmid yield of the P+ fruR cells was 21% higher than that of the P+ WT cells [19.2 versus 15.9 mg/g DCW (dry cell weight)]. These results demonstrate that, from an initial high-yielding fermentation process, the knockout of the fruR global regulator gene in E. coli DH5alpha further improves plasmid yields during fed-batch culture. PMID- 18380625 TI - HBeAg-negative chronic hepatitis B: cost-effectiveness of peginterferon alfa-2a compared to lamivudine in Taiwan. AB - OBJECTIVE: In Taiwan, the carrier rate of hepatitis B surface antigen is 15% to 20%, one of the highest in the world. Among chronic hepatitis B (CHB) patients, hepatitis B e antigen (HBeAg)-negative accounts for approximately 40% to 50% of these patients. A recent study found that peginterferon alfa-2a (40 KD) is more effective than lamivudine in treating HBeAg-negative CHB, but its cost effectiveness has not been evaluated. Our objective is to evaluate the incremental cost-effectiveness of 48 weeks of peginterferon alfa-2a compared to 48 weeks of lamivudine, from the perspective of the Taiwan Bureau of National Health Insurance. METHODS: A Markov model was used to simulate the natural history of HBeAg-negative CHB in a cohort of 40-year-old patients. Efficacy, disease progression, economic, and quality-of-life data were derived from published literature and a survey of clinical experts in Taiwan. Life expectancy, quality-adjusted life expectancy, lifetime costs in New Taiwan Dollars (NTD) (1 USD = 31.96 NTD), and incremental cost-effectiveness ratios (ICERs) were calculated. RESULTS: The gain in quality-adjusted life-years (QALYs) for 48 weeks of peginterferon alfa-2a compared to 48 weeks of lamivudine was 0.45 at an additional cost of 157,000 NTD (4900 USD), resulting in an ICER of 347,000 NTD (10,900 USD) per QALY gained. The 95% central range for the ICER from a probabilistic sensitivity analysis was 228,000-566,000 NTD (7100-17,700 USD). CONCLUSIONS: In HBeAg-negative CHB, 48 weeks of treatment with peginterferon alfa 2a compared to 48 weeks of lamivudine appears to offer life expectancy and quality-of-life improvements at an acceptable cost-effectiveness ratio. PMID- 18380626 TI - Probabilistic cost-effectiveness modeling of different breast cancer screening policies in Slovenia. AB - OBJECTIVES: To determine the most cost-effective screening policy for population based mammography breast cancer screening in Slovenia using probabilistic sensitivity analysis. METHODS: A time-dependent Markov model for breast cancer was constructed. General principles of cost-effectiveness analysis with multiple strategies were used to compare the costs and effects of 36 different screening policies. Using probability distributions for model parameters, the true effect of uncertainty across model input parameters on expected costs and effects was explored. The results from probabilistic simulation analysis are presented in a form of cost-effectiveness acceptability curves with cost-effectiveness acceptability frontier. RESULTS: With the presented analysis, it was shown that a 1-year screening interval in population breast cancer screening would produce less benefits at higher costs than less intensive screening and that a 2-year interval would be cost-effective only at high values of society's willingness to pay per quality-adjusted life-year (QALY). Therefore, the optimal screening policy should be chosen among 3-year-interval policies. CONCLUSIONS: Based on commonly quoted thresholds of society's willingness to pay per QALY of $50,000, the optimal approach in the Slovenian population would be screening women aged from 40 to 80 years every 3 years. PMID- 18380627 TI - Cost-effectiveness of ritodrine and fenoterol for treatment of preterm labor in a low-middle-income country: a case study. AB - OBJECTIVES: In countries with high income, tocolytic therapy with beta-mimetic agents is a cost-effective strategy compared to placebo. In our study, the cost effectiveness of two beta-mimetic agents, ritodrine and fenoterol, used in the management of preterm labor was compared in the setting of a low-middle-income transitional country, Serbia & Montenegro. METHODS: This case study was conducted at the Gynecology-Obstetrics Clinic, Clinical Center "Kragujevac," in Kragujevac, Serbia & Montenegro, between October 2004 and January 2006. In total, 235 pregnant patients with threatened preterm labor were enrolled, but 35 were lost to follow-up. Of the remaining 200 patients, 85 were given ritodrine, and 115 fenoterol. The perspective of Republic Institute for Health Insurance in Serbia was taken into account. Only direct costs were calculated; primary outcomes of the study were length of pregnancy (in weeks), time passed from the onset of uterine contractions to delivery (in weeks), and score on modified Flanagan's quality-of-life scale for chronic diseases, measured after discharge from hospital. RESULTS: Prolongation of pregnancy was significantly longer in the fenoterol group (12.7 +/- 8.4 weeks) than in the ritodrine group (11.6 +/- 7.1 weeks). The mean duration of hospitalization was shorter in the fenoterol group (11.9 +/- 8.8 days) than in the ritodrine group (14.9 +/- 11.3 days). The treatment with fenoterol was less costly and more cost-effective than the treatment with ritodrine, but the difference in cost-effectiveness was not statistically significant. The cost of treatment per gained week of pregnancy prolongation was 3345.51 +/- 7668.04 CSD in the fenoterol group, and 4181.96 +/- 12,069.83 CSD in the ritodrine group. CONCLUSIONS: The observed differences in treatment costs and duration of hospitalization per patient did not translate into significant differences in cost-effectiveness ratios, because of low costs of hospitalization and human labor in Serbian health system. Nevertheless, fenoterol treatment still has a tendency to be more cost-effective, and its lower acquisition cost is an advantage to this treatment option. PMID- 18380628 TI - Cost-effectiveness of rosuvastatin in the prevention of ischemic heart disease in Portugal. AB - OBJECTIVE: To assess the cost-effectiveness of rosuvastatin in the treatment of hypercholesterolemia and the prevention of ischemic heart disease (IHD) in Portugal. METHODS: A probabilistic Markov model was constructed to analyze the costs and consequences of lifetime treatment with rosuvastatin, atorvastatin, pravastatin, and simvastatin. For this purpose, the results from randomized head to-head trials evaluating low-density lipoprotein (LDL) changes were combined with the results from a meta-analysis defining the relationship between LDL levels and fatal and nonfatal IHD events. The incidence of myocardial infarction (MI) was derived from a 9-year Portuguese observational study. The eligible population was defined as untreated individuals aged more than 35 years with LDL levels above 115 mg/dl. Death rates due to IHD and other causes were obtained from official data. Resource use in the treatment of MI was estimated by a Delphi panel of eight Portuguese cardiologists with at least 15 years of clinical practice. Costs were calculated from the payers' perspective. RESULTS: Rosuvastatin increases life expectancy between 5.5 and 12.1 days per patient. It is cost-saving when compared to atorvastatin, but it increases costs when assessed against pravastatin and simvastatin (euro1,004 and euro684 per patient, respectively). Therefore, rosuvastatin is a dominant alternative compared to atorvastatin, having an incremental cost-effectiveness ratio of euro30,350 to pravastatin and euro39,340 to simvastatin. In the probabilistic sensitivity analysis, performed rosuvastatin always dominates atorvastatin and is associated with a cost per life-year gained inferior to euro50,000 in 95.7% of the cases when compared to pravastatin and in 67.0% simulations when assessed against simvastatin. CONCLUSIONS: Rosuvastatin is a cost-effective alternative in the prevention of IHD in Portugal. PMID- 18380629 TI - Cost-effectiveness and value of information analyses of neuraminidase inhibitors for the treatment of influenza. AB - OBJECTIVES: To assess the cost-effectiveness of alternative strategies for the treatment of suspected influenza in otherwise healthy adults and to identify future research priorities using value of information analysis. METHODS: A decision model was used to estimate the costs and effects, in terms of quality adjusted life-years (QALYs) of amantadine, zanamivir, and oseltamivir for the treatment of influenza in otherwise healthy adults using data predominantly from meta-analysis of randomized controlled trials. Probabilistic sensitivity analysis using Monte Carlo simulation was conducted. The expected value of perfect information for the entire model and for individual parameters was calculated. RESULTS: Based on mean costs and effects, zanamivir is dominated by oseltamivir. The incremental cost-effectiveness ratio for amantadine (compared with no treatment) is pound 11,000 and pound 44,000 for oseltamivir (compared with amantadine). The probability that amantadine is cost-effective at a willingness to pay of pound 30,000 per QALY is 0.74, falling to 0.49 at pound 20,000 per QALY. Global expected value of perfect information (EVPI) is pound 2 m over 15 years if a willingness to pay threshold of pound 30,000 per QALY is assumed rising to pound 9.6 m at pound 45,000 per QALY. EVPI for only one parameter exceeds pound 500,0000 at pound 30,000 per QALY: the quality of life for untreated influenza. CONCLUSIONS: At traditionally accepted values of willingness to pay for health benefits, it is unlikely that additional research would be an efficient use of scarce resources. The only exception to this would be to examine the health-related quality of life impact of influenza in an untreated patient group. If a higher threshold value were acceptable, there are a small group of parameters that may warrant further investigation. These would, however, require comparative, potentially expensive, research studies. PMID- 18380630 TI - Economic analysis of prophylactic pegfilgrastim in adult cancer patients receiving chemotherapy. AB - OBJECTIVES: Neutropenia and its complications, including febrile neutropenia (FN), are a common side effect of cancer chemotherapy. Results of clinical trials showed that prophylactic use of granulocyte colony-stimulating factors (G-CSF) is effective in preventing FN. In this study, the cost effectiveness (measured as cost per quality-adjusted time [days]) of three treatment alternatives were evaluated: no G-CSF, filgrastim administered daily for 7-12 days after chemotherapy, and a pegylated form of G-CSF pegfilgrastim, administered once per cycle. METHODS: A cost-utility model based on standard clinical practice of treating FN with immediate hospitalization or with ambulatory treatment, from a societal perspective was developed. Direct medical cost estimates for hospitalization were derived from claims data reported by 115 US academic medical centers. Indirect medical costs, productivity costs, probabilities, and utilities are based on published literature. Results were subjected to sensitivity analyses and 95% confidence intervals are based on a Monte Carlo simulation. RESULTS: Mean estimated costs/day of hospitalization were $1984 (SD $1040, N = 24,687) for surviving patients and $3139 (SD $2014, N = 1437) for dying patients. Under baseline conditions, pegfilgrastim dominated both filgrastim and no G-CSF, with expected costs and effectiveness of $4203 and 12.361 quality adjusted life-days (QALDs) for no G-CSF, $3058 and 12.967 QALDs for pegfilgrastim, and $5264 and 12.698 QALDs for filgrastim. CONCLUSIONS: This cost-utility analysis provides strong evidence that pegfilgrastim is not only cost-effective but also cost saving in most common clinical and economic settings. There appear to be both clinical and economic benefits from prophylactic administration of pegfilgrastim. PMID- 18380631 TI - Estimating the cost of complications of diabetes in Australia using administrative health-care data. AB - OBJECTIVES: To estimate Australian health-care costs in the year of first occurrence and subsequent years for major diabetes-related complications using administrative health-care data. METHODS: The costs were estimated using administrative information on hospital services and primary health-care services financed through Australia's national health insurance system Medicare. Data were available for 70,340 patients with diabetes in Western Australia (mean duration of 4.5 years of follow-up). Multiple regression analysis was used to estimate inpatient and primary care costs. RESULTS: For a man aged 60 years, the average costs in the year the event first occurred were: amputation $20,416 (95% CI 18,670-22,411); nonfatal myocardial infarction (MI) $11,660 (10,931-12,450); nonfatal stroke $14,012 (12,849-15,183); ischaemic heart disease $12,577 (12,026 13,123); heart failure $15,530 (13,965-17,009); renal failure $28,661 (22,989 34,202); and chronic leg ulcer $15,413 (13,089-18,123). The costs in subsequent years for a man aged 60 years range from 14% for nonfatal MI to 106% for renal failure, of event costs. CONCLUSIONS: Estimates of the health-care costs associated with diabetes-related complications can be used in modeling the long term costs of diabetes and in evaluating the cost-effectiveness of improving care. PMID- 18380632 TI - A multinational investigation of time and traveling costs in attending anticoagulation clinics. AB - OBJECTIVES: Anticoagulation is used in patients with atrial fibrillation to reduce the risk of ischemic stroke. The therapy requires regular monitoring and, frequently, dose adjustment. This study aimed to determine the time and traveling costs that patients incur to themselves and society in attending anticoagulation clinics. METHODS: A subset of patients from 105 primary and secondary care clinics allocated to the warfarin arm of SPORTIF III (patients from Australia, France, Portugal, Spain, Sweden, and the UK) completed a questionnaire. Patients indicated the type of transport used for clinic visits, and estimated traveling expenses. Patients were also asked to estimate total traveling and clinic attendance time, and to confirm whether they were currently employed and whether they had to give up time from work to attend the clinic. Time cost of companions was also taken into consideration. Cost per visit was calculated (euro, 2003 prices). RESULTS: Questionnaires for a total of 381 patients were analyzed, with the majority of patients from Sweden (n = 130) and the UK (n = 101). Mean cost to patients varied widely between countries, ranging from euro6.9 (France) to euro20.5 (Portugal) per visit. For most countries, time costs (value of lost working and leisure time) were the main driver of costs. Mean time cost to society ranged from euro5.6 (France) to euro31.7 (Portugal) per visit. CONCLUSIONS: Patients incur considerable costs when visiting anticoagulation clinics, and these costs vary by country. The results suggest the importance of taking a broad economic perspective when considering the cost-effectiveness of warfarin. PMID- 18380633 TI - Ten-year survival and cost following breast cancer recurrence: estimates from SEER-medicare data. AB - OBJECTIVE: A variety of pharmacologic therapies are available or in development for the prevention of breast cancer recurrence. Assessing the value of these treatments is compromised by a paucity of data on the impact of recurrence on economic costs and survival. The purpose of this study was to shed light on these issues. METHODS: We conducted a retrospective analysis of linked SEER-Medicare data. All patients in the National Cancer Institute's Surveillance Epidemiology and End Results (SEER) registry who were diagnosed with nonmetastatic breast cancer during 1991-1993 were identified, and their subsequent Medicare claims were scanned for evidence of further breast cancer events (local or distant recurrence, contralateral breast cancer). Medicare claims were then scanned from the time of the event through 2002 to assess patterns of survival and costs. RESULTS: We identified 10,798 patients in SEER who were diagnosed with nonmetastatic breast cancer during 1991-1993, including 1833 who subsequently had another breast cancer event (local recurrence, 958; distant recurrence, 622; contralateral breast cancer, 253). Median survival was 37 months and 8 months among patients with local and distant recurrence, respectively; 53% of patients with contralateral breast cancer remained alive after all the data were censored at 97 months. Expected 10-year costs (2004 US$, discounted 3%) attributable to distant recurrence, local recurrence, and contralateral breast cancer were $11,450 (SE 2056), $19,596 (SE 1754), and $19,183 (SE 4131), respectively. CONCLUSION: Breast cancer recurrence and contralateral breast cancer lead to substantial increases in costs, amounting to approximately $11,000-19,000 over 10 years depending on type. The impact of these events on survival also varies considerably by type. PMID- 18380634 TI - Costs associated with diffuse large B-cell lymphoma patient treatment in a Canadian integrated cancer care center. AB - OBJECTIVE: The objective of this study was to provide a detailed comparative microcosting analysis for two cancer treatment pathways to contribute evidence for resource allocation and operational decision-making in a Canadian cancer care context. METHODS: We estimated direct medical costs (in 2004 CAN$) of the entire pathway of care for diffuse large B-cell lymphoma (DLBCL) patients in a large Canadian cancer treatment center. Patient samples were defined as those who received R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone; n = 85) or CHOP (i.e., without rituximab; n = 86) as first-line treatment. All subsequent treatments including palliative care for these patient samples were assessed. Hospitalization costs and unit costs of medical resources were collected from integrated medical organizations. Individual patient resource consumption was assessed via medical chart review. RESULTS: For first-line treatment, drug cost was the largest contributor to total cost, followed by hospitalization cost. Rituximab was the largest contributor to mean cost differences between R-CHOP and CHOP treatments. For treatments subsequent to first-line treatment, no significant cost differences were found. Hospitalization and transplantation costs were the two largest constituents of total costs subsequent to first-line treatment, followed by drug cost. Patients with advanced stage disease cost significantly more than patients with limited stage disease. CONCLUSION: This is the first detailed microcosting study that has employed consistent local data to estimate total medical costs for DLBCL patients in Canada. This information is useful for resource allocation planning and operational decisions, because it provides more substantive, relevant evidence as compared to aggregated, literature or extrapolated information. PMID- 18380635 TI - Assessing productivity loss and activity impairment in severe or difficult-to treat asthma. AB - OBJECTIVES: Asthma can be associated with substantial productivity loss and activity impairment, particularly among those with the most severe disease. We sought to assess the performance characteristics of an asthma-specific adaptation of the Work Productivity and Activity Impairment Questionnaire (WPAI:Asthma) in patients with either severe or difficult-to-treat asthma. METHODS: We analyzed 2529 subjects from The Epidemiology and Natural History of Asthma: Outcomes and Treatment Regimens (TENOR) study. The WPAI:Asthma was administered at baseline and at 12 months. Asthma control and quality-of-life were simultaneously assessed using the Asthma Therapy Assessment Questionnaire and Mini-Asthma Quality-of-Life Questionnaire, respectively. RESULTS: Severe versus mild-to-moderate asthma was associated with a greater percentage of impairment at work (28% vs. 14%), at school (32% vs. 18%), and in daily activities (41% vs. 21%). At baseline, greater asthma control problems correlated with higher levels of impairment as measured by the WPAI (work: r = 0.54, school: r = 0.37, activity: r = 0.55). Over the 12 month follow-up period, improved quality-of-life correlated with decreased levels of impairment (work: r = -0.42, school: r = -0.36, activity: r = -0.48). In multivariate analyses, greater than 10% overall work impairment at baseline predicted emergency visits (OR 2.6 [1.6, 4.0]) and hospitalization (OR 4.9 [1.8, 13.1]) at 12 months. CONCLUSIONS: The WPAI:Asthma correlates with other self reported asthma outcomes in the expected manner and predicts health-care utilization at 12 months when administered to patients with severe or difficult to-treat asthma. PMID- 18380636 TI - Gaussian process metamodeling in Bayesian value of information analysis: a case of the complex health economic model for breast cancer screening. AB - OBJECTIVES: To determine whether allocation of resources into further research of breast cancer screening is warranted; also, to identify the parameters, for which the information would be most valuable, to prioritize the further research if deemed justifiable. METHODS: The Bayesian value of information analysis was conducted to calculate the overall expected value of perfect information (EVPI) and the partial EVPI for the six groups of parameters. Computational expense of the partial EVPI calculation was challenged with the use of Multiple Linear Regression and Gaussian Process metamodels to significantly cut down the computing time. RESULTS: Of the two metamodeling techniques, the Gaussian Process was proven to perform superiorly and was therefore chosen for the partial EVPI calculation. The results indicate a considerable range in the population EVPI estimates, between euro100 and euro500 millions at the willingness-to-pay values between euro10,000 and euro40,000 per quality-adjusted life-year. The partial EVPI for the groups of parameters indicated that future research would be most valuable if directed toward obtaining more precise estimates of the cancer sojourn times. With the use of the Gaussian process metamodels, the computing time was reduced from 44 years to 47 days. CONCLUSIONS: Although the large values of EVPI suggest collection of further information before choosing the screening policy, it is argued that delaying the decision would result in significantly higher opportunity loss. Therefore, the best option would be to implement the most cost-effective policy given the existing information (screening women aged 40-80 years, at 3-year intervals) and simultaneously conduct observational studies alongside the implemented policy. The decision analytic model could be in this manner periodically updated with additional information as it became available and the most cost-effective policy chosen iteratively. PMID- 18380637 TI - Practical guidelines for economic evaluations alongside equivalence trials. AB - An effective treatment already exists for many diseases. In these cases the effectiveness of a new treatment may be established by showing that the new treatment is as effective as (i.e., equivalent to) or at least as effective as (i.e., noninferior to) the old treatment. For an economic evaluation accompanying a clinical equivalence or noninferiority trial it is important to decide before the start of the study on the appropriate research question. In many cases the objective of the economic evaluation will be to show equivalence or noninferiority of the cost-effectiveness of the treatments. This has major implications for the design and analysis of the economic evaluation. In this article we propose methods for the analysis of economic equivalence and noninferiority studies that are similar to the methods applied to clinical equivalence and noninferiority trials. Furthermore, cost-effectiveness planes prove to be a valuable tool in the interpretation of the results in an economic equivalence or noninferiority trial. The concepts described in the article are illustrated using the results from an economic noninferiority trial. PMID- 18380638 TI - Advanced cutaneous malignant melanoma: a systematic review of economic and quality-of-life studies. AB - OBJECTIVE: Metastatic melanoma (MM), a major concern for health-care providers, is increasing. We systematically reviewed published articles describing the impact of interventions (drugs and screening) on quality of life (QoL) in patients with MM, and articles that measured QoL in MM. METHODS: We searched secondary databases including MEDLINE, Embase, CINAHL, Cochrane, and DARE from inception to 2006 using MESH terms "melanoma" and "metastases." Economic articles were subject to established quality assessment procedures. RESULTS: We found 13 QoL and five economic studies (three cost-effectiveness, two cost-utility; average quality = 83% +/- 7%). No strong evidence was found in this review for cost-effectiveness of interferons in Canada (incremental cost-effectiveness ratio [ICER] = $55,090/quality-adjusted life-year) or temozolomide in the United States (ICER = $36,990/Life-year gained based on nonsignificant efficacy differences). Melanoma screening was not cost-effective in the United States ($150,000 931,000/life-saved) or Germany (no survival benefit). From the 13 QoL studies,eight measured baseline QoL; six studied the same population, generating similar results using different approaches/outcomes. Tools used included GLQ-8, QLQ-C30, QLQ-36, QWB-SA, and SF-36. Baseline scores QoL scores ranged from 0.60 to 0.69. Another five studies (N = 959 patients) were randomized trials analyzing QoL in patients treated with dacarbazine alone, dacarbazine +/- interferon, dacarbazine + fotemustine, interleukin +/- histamine, and temozolomide. Little difference was found in QoL scores between drugs or between baseline and end point. CONCLUSIONS: Cost-effectiveness has not been widely demonstrated for treatment of MM. Only two studies with unimpressive results exist for treatments. Screening was not cost-effective in the United States or Germany. Generally, no significant improvements in QoL were found for any alternative for treating MM. A need exists for effective treatments that improve duration and QoL. PMID- 18380639 TI - Cost-utility of repeated screening for Chlamydia trachomatis. AB - OBJECTIVE: To estimate the cost-effectiveness of repeated screening for chlamydia trachomatis at various time intervals compared to one-off screening of Dutch young adults. METHODS: We used a dynamic model to fully take the spread of the disease over time in the population into account, with data being used gathered within the context of a recently performed pilot study in The Netherlands. The screening frequencies analyzed were: every year, every 2 years, every 5 years, and every 10 years. The strategies were compared in terms of incremental cost effectiveness, expressed as the net costs per quality-adjusted life-year (QALY). RESULTS: For all interval strategies, with the exception of screening every year, incremental cost-effectiveness stays below the informal Dutch threshold of euro20,000 per QALY. CONCLUSION: From a health-economic point of view, for the Dutch situation, we estimated screening every 2 years as the optimal strategy among the options investigated. PMID- 18380640 TI - Comparing the standard EQ-5D three-level system with a five-level version. AB - OBJECTIVES: The aim of this study is a head-to-head comparison of the performance of the three-level EQ-5D (3L) and a newly developed five-level version (5L). METHODS: Eighty-two respondents valued 15 standardized disease descriptions and their own health on three response scales (3L, 5L, and visual analog scale [VAS]) for all five EQ-5D dimensions. Performance was studied in terms of feasibility, face validity, redistribution properties, ordinality, convergent validity, discriminatory power, and test-retest and interobserver reliability. RESULTS: The majority of participants judged 5L as the preferred system in terms of feasibility (76%) and face validity (75%). In total, 1.1% of responses were inconsistent. Ordinality of 5L was confirmed in all cases. Convergent validity of 3L-VAS (range: 0.88-0.99) and 5L-VAS (0.90-0.99) were high and about equal. Discriminatory power (informativity) improves considerably with 5L without loss of Evenness. Interobserver reliability (0.49 vs. 0.57) and test-retest reliability (0.52 vs. 0.69) were higher in 5L. CONCLUSION: The EQ-5D five-level version appears a valid and reliable extension of the three-level system. The new 5L system is particularly useful for describing mild health problems and monitoring population health. PMID- 18380641 TI - Evaluation of new measures of the impact of hypothyroidism on quality of life and symptoms: the ThyDQoL and ThySRQ. AB - OBJECTIVES: This article reports the psychometric properties of two new condition specific questionnaires: 1) the 18-item Underactive Thyroid-Dependent Quality of Life Questionnaire (ThyDQoL) individualized measure of perceived impact of hypothyroidism on quality of life (QoL); and 2) the 15-item Underactive Thyroid Symptom Rating Questionnaire (ThySRQ), in which patients rate symptom bother. METHODS: A cross-sectional survey was conducted of 110 adults with overt and subclinical hypothyroidism, 103 treated with thyroxine. Patients, the majority of whom (81%) were women, were recruited from primary care (57%) and from hospital clinics (43%). The mean age of patients was 55.1 (SD 14.3) years. Respondents rated personally applicable ThyDQoL life domains for importance and impact of hypothyroidism, and ThySRQ symptom bother. RESULTS: Completion rates were high (>98%). All 18 ThyDQoL domains were rated as negatively impacted by hypothyroidism and important for QoL. The ThyDQoL had high internal consistency reliability (Cronbach's alpha = 0.949 [N = 97]), factor analysis indicated that applicable domains could be combined into an overall Average Weighted Impact score, for which the sample mean, -3.11 (2.2), showed considerable negative impact of hypothyroidism on QoL (maximum possible range -9 to +3). There is good preliminary evidence to justify shortening the ThyDQoL to 14 domain-specific items. For the ThySRQ Cronbach's alpha was 0.808 (N = 95). Highest symptom bother ratings were for hair problems, weight gain, depression, cold, and tiredness. CONCLUSIONS: Both the ThyDQoL and ThySRQ are highly acceptable to patients with hypothyroidism and have good internal consistency reliability. Their sensitivity to change now needs to be evaluated in clinical trials. PMID- 18380642 TI - Valuation of the SF-6D Health States Is Feasible, Acceptable, Reliable, and Valid in a Chinese Population. AB - OBJECTIVES: The SF-6D is a preference-based measure of health (PBMH) derived from the SF-36 for economic evaluation. The aim of this study was to find out whether it was feasible, acceptable, reliable, and valid to use the standard gamble (SG) method to generate preference-based values for the SF-6D in a Chinese population. METHODS: The SF-6D was translated into Chinese by forward and backward translations. Forty-nine states defined by the SF-6D were selected using an orthogonal design and grouped into seven sets. An age-sex stratified sample of 126 Chinese adults with low education levels valued a set of 7 and the pits (worst) SF-6D health states by the SG method. The data were modeled at the individual and mean levels to predict preference values for all SF-6D states. The quality of data and the predictive power of the models were compared with results from the United Kingdom. RESULTS: All respondents completed the interviews with 3% finding the process very difficult and 21% felt some degree of irritation or boredom. A total of 907 SG valuations (90% out of 1008 observations) were useable for econometric modeling. There was no significant change in the test-retest values from 21 subjects. The main mean effect models achieved a good fit with a mean absolute error of 0.054. Some differences between the Chinese and UK preference coefficients were found especially in the physical functioning dimension. The range of SG values predicted by the HK function is slightly longer, with the pits state having a value of 0.152 compared to 0.271 in the UK. CONCLUSION: It was feasible, acceptable, reliable, and valid to value the SF-6D with the SG method in a Chinese population with relatively low education levels. The results supported the feasibility and validity of valuing PBMH in Asian populations. Further studies are required to determine whether the differences in the SF-6D scoring algorithms between the British and Chinese populations are important. PMID- 18380643 TI - Estimating the effect of treatment rate changes when treatment benefits are heterogeneous: antibiotics and otitis media. AB - OBJECTIVE: Contrast methods to assess the health effects of a treatment rate change when treatment benefits are heterogeneous across patients. Antibiotic prescribing for children with otitis media (OM) in Iowa Medicaid is the empirical example. METHODS: Instrumental variable (IV) and linear probability model (LPM) are used to estimate the effect of antibiotic treatments on cure probabilities for children with OM in Iowa Medicaid. Local area physician supply per capita is the instrument in the IV models. Estimates are contrasted in terms of their ability to make inferences for patients whose treatment choices may be affected by a change in population treatment rates. RESULTS: The instrument was positively related to the probability of being prescribed an antibiotic. LPM estimates showed a positive effect of antibiotics on OM patient cure probability while IV estimates showed no relationship between antibiotics and patient cure probability. CONCLUSIONS: Linear probability model estimation yields the average effects of the treatment on patients that were treated. IV estimation yields the average effects for patients whose treatment choices were affected by the instrument. As antibiotic treatment effects are heterogeneous across OM patients, our estimates from these approaches are aligned with clinical evidence and theory. The average estimate for treated patients (higher severity) from the LPM model is greater than estimates for patients whose treatment choices are affected by the instrument (lower severity) from the IV models. Based on our IV estimates it appears that lowering antibiotic use in OM patients in Iowa Medicaid did not result in lost cures. PMID- 18380644 TI - Migraine frequency and health utilities: findings from a multisite survey. AB - OBJECTIVES: Assess the relationship between migraine frequency and health utility. METHODS: Patients aged >/=18 years diagnosed with episodic migraine were enrolled at three US sites representing varied models of health-care delivery. All subjects completed a questionnaire that included demographic and clinical information, a migraine-specific disability questionnaire, and the Health Utilities Index Mark 3 (HUI3). The HUI3 is a generic health status and health related quality-of-life measure. HUI3 health status data are translated into utility scores anchored by 0 (dead) and 1 (perfect health). RESULTS: The study enrolled 150 patients. The mean age was 44 years and 87% were female. Mean (+/ SD) monthly migraine frequency was 4.4 +/- 3.6, with 34% reporting 6 migraines per month. The mean (+/-SD) HUI3 score was 0.62 +/- 0.26. After controlling for study center, demographics, comorbidities, migraine characteristics, and level of migraine disruptiveness, migraine frequency was found to be significantly (P < 0.05) and negatively associated with HUI3 scores. Subjects with >6 migraines per month had an adjusted mean HUI3 score of 0.41; the corresponding mean for those reporting 18 000 Ae. aegypti and approximately 170 000 Cx quinquefasciatus adults per day. Septic tanks are likely to be common and widespread in suburban and rural Puerto Rico, where, apparently, they can contribute significantly to the maintenance of island-wide dengue virus endemicity. PMID- 18380656 TI - Survivorship characteristics of the mosquito Aedes caspius adults from southern France under laboratory conditions. AB - The survivorship characteristics of two populations of Aedes caspius (Pallas) (Diptera: Culicidae) were compared in the laboratory. One population was sourced from Mourgues, where larvicides have been used continuously for approximately 40 years, and the other from Pont de Gau, where there has been no consistent mosquito control. The aims of the study were to ascertain the basic life history profiles of adults and to determine whether continuous larviciding affects inherent adult survivorship. Life tables were constructed to calculate the following life expectancy parameters: mean lifetime (tau(ad)); maximum lifetime (tau(max)), and daily survival rate (p(ad)). All three parameters were higher for females than for males (paired t-test, P < or = 0.001); male mean lifetime, maximum lifetime and daily survival rate were 4.95 +/- 0.94 days, 20.50 +/- 6.66 days and 0.79 +/- 0.05, respectively; female values were 14.74 +/- 3.68 days, 49.69 +/- 16.55 days and 0.93 +/- 0.02, respectively. No differences were found between the two populations, and no correlations were found between initial adult densities and their respective survival rates. The survivorship curves for Ae. caspius were type IV for males (mortality rates higher for young adults) and type III for females (mortality rates constant). PMID- 18380657 TI - Potential impacts of climate change on stable flies, investigated along an altitudinal gradient. AB - Adult populations of stable flies were sampled along an altitudinal transect in Reunion Island to determine whether higher temperatures were associated with: (a) higher numbers of flies; (b) a longer season of infestation, and/or (c) different responses to warming in the cosmopolitan Stomoxys calcitrans (L) and the tropical Stomoxys niger niger Macquart (Diptera: Muscidae). Flies of both species were trapped at seven farms situated at four altitudes (100-1600 m a.s.l.) over a 90 week period. For both species, there were no relationships between the maximum or mean fly abundance and altitude. Only minimum abundance in winter was significantly higher at lower altitudes. Maximum and mean abundances differed significantly between nearby farms under similar climatic conditions. Seasonal fluctuations in fly abundance changed along the gradient. At lower altitudes, population growth started earlier after the winter but abundance declined earlier in summer, which resulted in a shift of the season of infestation. Seasonal fluctuations of both species were strongly related to climate variables at high altitude, mainly temperature. However, climate variables explained a decreasing proportion of the variations in abundance at lower altitudes. Stomoxys calcitrans was the most abundant species overall, but the proportion of S. n. niger increased significantly at lower altitudes and this species became predominant at 100 m a.s.l. It is concluded that stable fly infestations are unlikely to worsen in response to global warming. Maximum abundance is limited by local factors, possibly larval resources, which suggests that adequate husbandry practices could override the impact of climate change. Because S. n. niger tends to be the predominant pest at elevated temperatures, it is recommended that this species should not be introduced in areas where climate is changing. PMID- 18380658 TI - High-resolution melt analysis for the detection of a mutation associated with permethrin resistance in a population of scabies mites. AB - Permethrin as a topical acaricide cream is widely used to treat scabies. The neuronal voltage-sensitive sodium channel (Vssc), necessary for the generation of action potentials in excitable cells, is the target of pyrethroid acaricides such as permethrin. Pyrethroid resistance has been linked to specific mutations in the Vssc gene. Following the partial sequencing of the Vssc gene in the scabies mite Sarcoptes scabiei (L.) (Astigmata: Sarcoptidae), we compared Vssc gene sequences from permethrin-sensitive and -tolerant S. scabiei var. canis Gerlach mites, and identified a G to A single nucleotide polymorphism (SNP) in permethrin-tolerant mites resulting in an amino acid change from glycine to aspartic acid in domain III S6. The mutation is in a region of the gene where mutations have been identified in a range of pyrethroid-resistant arthropods. Results of in vitro permethrin exposure assays showed that survival rates for mites bearing the mutation were similar to those previously reported for mites from human subjects where clinical tolerance to permethrin had been observed. A real-time polymerase chain reaction-high-resolution melt (PCR-HRM) assay was developed to detect this SNP. This assay provides a useful methodology for screening for this and other mutations associated with permethrin resistance in scabies mite populations and thus facilitates surveillance for acaricide resistance. PMID- 18380659 TI - Mitochondrial genomes of the sheep blowfly, Lucilia sericata, and the secondary blowfly, Chrysomya megacephala. AB - This paper presents complete mitochondrial genomes for the sheep blowfly, Lucilia sericata (Meigen), and the secondary blowfly, Chrysomya megacephala (Fabricius). Both L. sericata and C. megacephala had standard dipteran-type mitochondrial genome architectures and lengths of 15 945 bp and 15 831 bp, respectively. Additionally, C. megacephala possessed a tRNA duplication either side of the D loop, as previously reported in another Chrysomya species, C. putoria; this duplication appears to be synapomorphic for the genus Chrysomya. As in other insect mitochondrial genomes, base compositions had a high AT content, with both genomes more than 76% AT-rich. PMID- 18380660 TI - Modelling hepatitis C virus kinetics during treatment with pegylated interferon alpha-2b: errors in the estimation of viral kinetic parameters. AB - Neumann et al. [1] developed a widely used model for the analysis of hepatitis C virus (HCV) dynamics after the initiation of interferon therapy that assumes the effectiveness of therapy in blocking virion production, epsilon, is constant. However, with pegylated interferon alpha-2b (PEG-IFN) given weekly, there are significant changes in drug concentration between doses, leading to changes in drug effectiveness and viral rebounds. To investigate the appropriateness of the constant effectiveness (CE) model [1] for studies involving PEG-IFN, we simulated PEG-IFN treatment, using 294 sets of pharmacokinetic/pharmacodynamic (PK/PD) parameters that span observed ranges and fit the simulated data to the CE model. For most combinations of PK/PD parameters, the fits resulted in an infected cell loss rate, delta, that underestimates the true value used in the simulations and yielded over-estimates of the average effectiveness of PEG-IFN. In the setting of PEG-IFN therapy, the use of the CE model of HCV kinetics has to be reevaluated and the validity of its use depends on the amount of HCV RNA rebound observed between doses. PMID- 18380662 TI - Endothelial cell-derived bone morphogenetic proteins regulate glial differentiation of cortical progenitors. AB - Gliogenesis is an important component of cortical development during the postnatal period. Two macroglial cells are generated in a particular order, i.e. astrocytes first and oligodendrocytes later. The mechanisms underlying this sequence of glial differentiation are unknown but interactions with blood vessels are postulated to play a role. We show, using a mouse in-vitro coculture system, that endothelial cells promote astrocyte differentiation but inhibit oligodendrocyte differentiation of postnatal cortical progenitors. Endothelial cells produce bone morphogenetic proteins (BMPs) to activate Sma- and Mad-related protein (Smad) signalling in progenitors and the effects of endothelial cells on glial differentiation are blocked by the BMP antagonist Noggin. Differentiation of progenitors into astrocytes results in the inhibition of endothelial cell growth, accompanied by changes in gene expression of angiogenic factors, indicating bidirectional interactions between progenitors and endothelial cells. In vivo, Smad signalling is activated in various types of cortical cells including progenitors in association with astrogenesis but is inactivated before the peak of oligodendrogenesis. Capillary vessels isolated from the developing cortex express high levels of BMPs. Together, these results demonstrate that endothelial cells regulate glial differentiation by secreting BMPs in vitro and suggest a similar role in cortical gliogenesis in vivo. PMID- 18380661 TI - ADAM2 promotes migration of neuroblasts in the rostral migratory stream to the olfactory bulb. AB - Neuroblasts migrate from the subventricular zone along the rostral migratory stream (RMS) to the olfactory bulb (OB). While the migration occurs by movement over other cells, the molecular mechanisms are poorly understood. We have found that ADAM2 (a disintegrin and metalloprotease 2) is expressed in migrating RMS neuroblasts and functions in their migration. The brains from ADAM2 knockout (KO) mice showed a smaller OB than that seen in wild-type (WT) mice at postnatal day 0. In addition, the RMS in ADAM2 KO mice appeared thinner and less voluminous in its rostral part and thicker in its caudal part. Estimates of migration in vivo using bromodeoxyuridine labeling revealed that neuroblasts from KO mice show a decreased migration rate compared with those from WT mice. Direct assays of migration by imaging living slices also showed a decreased migration speed and loss of directionality in the KO mice. This phenotype was similar to that seen in RMS containing slices from WT mice exposed to a peptide that mimicked the disintegrin loop of ADAM2. Finally, RMS explants from KO or WT mice that were cultured in Matrigel also revealed striking differences. The cells migrating out of explants from WT mice showed robust cell-cell interactions. In contrast, fewer cells migrated out of explants from ADAM2 KO mice, and those that did were largely dispersed and their migration inhibited. These experiments suggest that ADAM2 contributes to RMS migration, possibly through cell-cell interactions that mediate the rapid migration of the neuroblasts to their endpoint. PMID- 18380663 TI - Somal positioning and dendritic growth of horizontal cells are regulated by interactions with homotypic neighbors. AB - Retinal neurons extend their dendritic fields to achieve a degree of dendritic overlap with homotypic neighbors that is cell-type specific. How these neurons regulate their dendritic growth is unclear. The dendritic field of a retinal horizontal cell varies inversely with horizontal cell density across different strains of mice, suggesting that proximity to neighboring cells regulates dendritic growth. To test this directly, we have employed the Cre-loxP conditional gene targeting strategy to achieve inactivation of Lim1 function in developing horizontal cells. Through this approach, Lim1 function was prevented within a subset of horizontal cells that in turn fail to migrate to the horizontal cell layer and differentiate normally. For those remaining horizontal cells with Lim1 intact (about half of the normal population in these mice), we show that they spread themselves out tangentially and differentiate a dendritic morphology that is essentially normal but for the fact that it has nearly doubled in area. Such larger horizontal cells, sampling from an area of retina containing twice their normal afferent number, differentiate a dendritic field with nearly double the number of higher order branches and terminal clusters. These results demonstrate directly that positioning and dendritic growth are regulated by interactions with homotypic neighbors, whereas afferents instruct the differentiation of dendritic patterning. PMID- 18380665 TI - ALG-2 interacting protein AIP1: a novel link between D1 and D3 signalling. AB - Dopamine signalling is a critically important process in the human brain that controls mood, cognition and motor activity. In order to gain detailed insight into this signalling pathway at the molecular level, we carried out yeast two hybrid screens with D1-like (D1, D5) and D2-like (D2, D3, D4) dopamine receptors and identified 11 dopamine receptor interacting proteins (DRIPs). Using the C terminal domain of D1 receptor as bait, we identified AIP1 (ALG-2 interacting protein 1), a known modulator of caspase-dependent and caspase-independent cell death, including neuronal cell death, that is also part of the endosomal transport system. In a separate yeast two-hybrid screen, using the third intracellular cytoplasmic loop of D3 as bait, we again identified AIP1. The interaction of AIP1 with both D1 and D3 was confirmed in vitro and in vivo using a variety of methods, including glutathione S-transferase (GST) pull-down, blot overlay and coimmunoprecipitation from mouse brain lysates. We have also observed colocalization of D1 and D3 with AIP1 in mouse brain tissue. In addition, coexpression of AIP1 with D1 resulted in > 50% reduction in binding capacity of D1 to its antagonist. Finally, AIP1 up-regulates D1 and D3 expression and appears to be important for their stability and trafficking. PMID- 18380664 TI - Tau expression levels from various adeno-associated virus vector serotypes produce graded neurodegenerative disease states. AB - Neurodegenerative diseases involving neurofibrillary tangle pathology are pernicious. By expressing the microtubule-associated protein tau, a major component of tangles, with a viral vector, we induce neuropathological sequelae in rats that are similar to those seen in human tauopathies. We tested several variants of the adeno-associated virus (AAV) vector for tau expression in the nigrostriatal system in order to develop models with graded onset and completeness. Whereas previous studies with AAV2 tau vectors produced partial lesions of the nigrostriatal system, AAV9 or AAV10 tau vectors were more robust. These vectors had formidable efficacy relative to 6-hydroxydopamine for dopamine loss in the striatum. Time-courses for tau transgene expression, dopamine loss and rotational behavior tracked the disease progression with the AAV9 tau vector. There was a nearly complete lesion over a delayed time-course relative to 6 hydroxydopamine, with a sequence of tau expression by 1 week, dopamine loss by 2 weeks and then behavior effect by 3-4 weeks. Relative to AAV2 or AAV8, tau expression from AAV9 or AAV10 peaked earlier and caused more dopamine loss. Varying vector efficiencies produced graded states of disease up to nearly complete. The disease models stemming from the AAV variants AAV9 or AAV10 may be useful for rapid drug screening, particularly for tau diseases that affect the nigrostriatal system, such as progressive supranuclear palsy. PMID- 18380666 TI - Differential synaptic plasticity of the corticostriatal and thalamostriatal systems in an MPTP-treated monkey model of parkinsonism. AB - Two cardinal features of Parkinson's disease (PD) pathophysiology are a loss of glutamatergic synapses paradoxically accompanied by an increased glutamatergic transmission to the striatum. The exact substrate of this increased glutamatergic drive remains unclear. The striatum receives glutamatergic inputs from the thalamus and the cerebral cortex. Using vesicular glutamate transporters (vGluTs) 1 and 2 as markers of the corticostriatal and thalamostriatal afferents, respectively, we examined changes in the synaptology and relative prevalence of striatal glutamatergic inputs in methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated monkeys using electron microscopic immunoperoxidase and confocal immunofluorescence methods. Our findings demonstrate that the prevalence of vGluT1-containing terminals is significantly increased in the striatum of MPTP treated monkeys (51.9 +/- 3.5% to 66.5 +/- 3.4% total glutamatergic boutons), without any significant change in the pattern of synaptic connectivity; more than 95% of vGluT1-immunolabeled terminals formed axo-spinous synapses in both conditions. In contrast, the prevalence of vGluT2-immunoreactive terminals did not change after MPTP treatment (21.7 +/- 1.3% vs. 21.6 +/- 1.2% total glutamatergic boutons). However, a substantial increase in the ratio of axo spinous to axo-dendritic synapses formed by vGluT2-immunoreactive terminals was found in the pre-caudate and post-putamen striatal regions of MPTP-treated monkeys, suggesting a certain degree of synaptic reorganization of the thalamostriatal system in parkinsonism. About 20% of putative glutamatergic terminals did not show immunoreactivity in striatal tissue immunostained for both vGluT1 and vGluT2, suggesting the expression of another vGluT in these boutons. These findings provide striking evidence that suggests a differential degree of plasticity of the corticostriatal and thalamostriatal system in PD. PMID- 18380667 TI - Ammonium alters creatine transport and synthesis in a 3D culture of developing brain cells, resulting in secondary cerebral creatine deficiency. AB - Hyperammonemic disorders in pediatric patients lead to poorly understood irreversible effects on the developing brain that may be life-threatening. We showed previously that some of these NH4+-induced irreversible effects might be due to impairment of axonal growth that can be protected under ammonium exposure by creatine co-treatment. The aim of the present work was thus to analyse how the genes of arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT), allowing creatine synthesis, as well as of the creatine transporter SLC6A8, allowing creatine uptake into cells, are regulated in rat brain cells under NH4+ exposure. Reaggregated brain cell three-dimensional cultures exposed to NH4Cl were used as an experimental model of hyperammonemia in the developing central nervous system (CNS). We show here that NH4+ exposure differentially alters AGAT, GAMT and SLC6A8 regulation, in terms of both gene expression and protein activity, in a cell type-specific manner. In particular, we demonstrate that NH4+ exposure decreases both creatine and its synthesis intermediate, guanidinoacetate, in brain cells, probably through the inhibition of AGAT enzymatic activity. Our work also suggests that oligodendrocytes are major actors in the brain in terms of creatine synthesis, trafficking and uptake, which might be affected by hyperammonemia. Finally, we show that NH4+ exposure induces SLC6A8 in astrocytes. This suggests that hyperammonemia increases blood brain barrier permeability for creatine. This is normally limited due to the absence of SLC6A8 from the astrocyte feet lining microcapillary endothelial cells, and thus creatine supplementation may protect the developing CNS of hyperammonemic patients. PMID- 18380668 TI - Generation recruitment and death of brain cells throughout the life cycle of Sorex shrews (Lipotyphla). AB - Young shrews of the genus Sorex that are born in early summer reduce their body size before wintering, including a reduction of brain weight of 10-30%. In the spring they mature sexually, double their body weight and regain about half of the loss in brain weight. To investigate the mechanisms of brain weight oscillations we studied the rate of cell death and generation in the brain during the whole life cycle of the common shrew (Sorex araneus) and pygmy shrew (S. minutus). After weaning, shrews generate new brain cells in only two mammalian neurogenic zones and approximately 80% of these develop into neurones. The increase of the shrew brain weight in the spring did not depend on recruitment of new cells. Moreover, adult Sorex shrews did not generate new cells in the dentate gyri. Injections of 5-HT1A receptor agonists in the adult shrews induced neurogenesis in their dentate gyri, showing the presence of dormant progenitor cells. Generation of new neurones in the subventricular zone of the lateral ventricles and their recruitment to olfactory bulbs continued throughout life. TUNEL labelling showed that the rate of cell death in all brain structures, including the proliferation zones and olfactory bulb, was very low throughout life. We conclude that neither cell death nor recruitment significantly contributes to seasonal oscillations and the net loss of brain weight in the Sorex shrews. With the exception of dentate gyrus and olfactory bulb, cellular populations of brain structures are stable throughout the life cycle of these shrews. PMID- 18380669 TI - Evidence for a novel functional role of cannabinoid CB(2) receptors in the thalamus of neuropathic rats. AB - Cannabinoid CB(1) receptors have analgesic effects in models of neuropathic pain, but can also produce psychoactive side-effects. A supraspinal location of CB(2) receptors has recently been described. CB(2) agonists are also antinociceptive, although the functional role of supraspinal CB(2) receptors in the control of nociception is unknown. Herein, we provide evidence that CB(2) receptors in the thalamus play a functional role in the modulation of responses of neurons in the ventral posterior nucleus (VPL) of the thalamus in neuropathic, but not sham operated, rats. Spontaneous and mechanically evoked activity of VPL neurons was recorded with a multichannel electrode array in anaesthetized spinal nerve ligated (SNL) rats and compared to sham-operated rats. Intra-VPL administration of the CB(2) agonist JWH-133 (30 ng in 500 nL) significantly reduced spontaneous (P < 0.05), non-noxious (P < 0.001) and noxious (P < 0.01) mechanically evoked responses of VPL neurons in SNL rats, but not in sham-operated rats. Inhibitory effects of JWH-133 on spontaneous (P < 0.01) and noxious-evoked (P < 0.001) responses of neurons were blocked by the CB(2) antagonist SR144528. Local administration of SR144528 alone did not alter spontaneous or evoked responses of VPL neurons, but increased burst activity of VPL neurons in SNL rats. There were, however, no differences in levels of the endocannabinoids anandamide and 2AG in the thalamus of SNL and sham-operated rats. These data suggest that supraspinal CB(2) receptors in the thalamus may contribute to the modulation of neuropathic pain responses. PMID- 18380670 TI - Differential tonic influence of lateral habenula on prefrontal cortex and nucleus accumbens dopamine release. AB - Conditions of increased cognitive or emotional demand activate dopamine release in a regionally selective manner. Whereas the brief millisecond response of dopamine neurons to salient stimuli suggests that dopamine's influence on behaviour may be limited to signalling certain cues, the prolonged availability of dopamine in regions such as the prefrontal cortex and nucleus accumbens is consistent with the well described role of dopamine in maintaining motivation states, associative learning and working memory. The behaviourally elicited terminal release of dopamine is generally attributed to increased excitatory drive on dopamine neurons. Our findings here, however, indicate that this increase may involve active removal of a tonic inhibitory control on dopamine neurons exerted by the lateral habenula (LHb). Inhibition of LHb in behaving animals transiently increased dopamine release in the prefrontal cortex, nucleus accumbens and dorsolateral striatum. The inhibitory influence was more pronounced in the nucleus accumbens and striatum than in the prefrontal cortex. This pattern of regional dopamine activation after LHb inhibition mimicked conditions of reward availability but not increased cognitive demand. Electrical or chemical stimulation of LHb produced minimal reduction of extracellular dopamine, suggesting that in an awake brain the inhibition associated with tonic LHb activity represents a near-maximal influence on dopamine neurotransmission. These data indicate that LHb may be critical for functional differences in dopamine neurons by preferentially modulating dopamine neurons that project to the nucleus accumbens over those neurons that primarily project to the prefrontal cortex. PMID- 18380672 TI - Sleep architecture of the melanin-concentrating hormone receptor 1-knockout mice. AB - Growing amounts of data indicate involvement of the posterior hypothalamus in the regulation of sleep, especially paradoxical sleep (PS). Accordingly, we previously showed that the melanin-concentrating hormone (MCH)-producing neurons of the rat hypothalamus are selectively activated during a PS rebound. In addition, intracerebroventricular infusion of MCH increases total sleep duration, suggesting a new role for MCH in sleep regulation. To determine whether activation of the MCH system promotes sleep, we studied spontaneous sleep and its homeostatic regulation in mice with deletion of the MCH-receptor 1 gene (MCH-R1-/ vs. MCH-R1+/+) and their behavioural response to modafinil, a powerful antinarcoleptic drug. Here, we show that the lack of functional MCH-R1 results in a hypersomniac-like phenotype, both in basal conditions and after total sleep deprivation, compared to wild-type mice. Further, we found that modafinil was less potent at inducing wakefulness in MCH-R1-/- than in MCH-R1+/+ mice. We report for the first time that animals with genetically inactivated MCH signaling exhibit altered vigilance state architecture and sleep homeostasis. This study also suggests that the MCH system may modulate central pathways involved in the wake-promoting effect of modafinil. PMID- 18380673 TI - Addition of fornix transection to frontal-temporal disconnection increases the impairment in object-in-place memory in macaque monkeys. AB - Both frontal-inferotemporal disconnection and fornix transection (Fx) in the monkey impair object-in-place scene learning, a model of human episodic memory. If the contribution of the fornix to scene learning is via interaction with or modulation of frontal-temporal interaction--that is, if they form a unitary system--then Fx should have no further effect when added to frontal-temporal disconnection. However, if the contribution of the fornix is to some extent distinct, then fornix lesions may produce an additional deficit in scene learning beyond that caused by frontal-temporal disconnection. To distinguish between these possibilities, we trained three male rhesus monkeys on the object-in-place scene-learning task. We tested their learning on the task following frontal temporal disconnection, achieved by crossed unilateral aspiration of the frontal cortex in one hemisphere and the inferotemporal cortex in the other, and again following the addition of Fx. The monkeys were significantly impaired in scene learning following frontal-temporal disconnection, and furthermore showed a significant increase in this impairment following the addition of Fx, from 32.8% error to 40.5% error (chance = 50%). The increased impairment following the addition of Fx provides evidence that the fornix and frontal-inferotemporal interaction make distinct contributions to episodic memory. PMID- 18380671 TI - Role for the NR2B subunit of the N-methyl-D-aspartate receptor in mediating light input to the circadian system. AB - Light information reaches the suprachiasmatic nucleus (SCN) through a subpopulation of retinal ganglion cells that utilize glutamate as a neurotransmitter. A variety of evidence suggests that the release of glutamate then activates N-methyl-D-aspartate (NMDA) receptors within the SCN and triggers a signaling cascade that ultimately leads to phase shifts in the circadian system. In this study, we first sought to explore the role of the NR2B subunit in mediating the effects of light on the circadian system of hamsters and mice. We found that localized microinjection of the NR2B subunit antagonist ifenprodil into the SCN region reduces the magnitude of light-induced phase shifts of the circadian rhythm in wheel-running activity. Next, we found that the NR2B message and levels of phospho-NR2B vary with time of day in SCN tissue using semiquantitative real-time polymerase chain reaction and western blot analysis, respectively. Functionally, we found that blocking the NR2B subunit with ifenprodil significantly reduced the magnitude of NMDA currents recorded in SCN neurons. Ifenprodil also significantly reduced the magnitude of NMDA-induced Ca2+ changes in SCN cells. Together, these results demonstrate that the NR2B subunit is an important component of NMDA receptor-mediated responses within SCN neurons and that this subunit contributes to light-induced phase shifts of the mammalian circadian system. PMID- 18380674 TI - Learning-related changes in reward expectancy are reflected in the feedback related negativity. AB - The feedback-related negativity (FRN) has been hypothesized to be linked to reward-based learning. While many studies have shown that the FRN only occurs in response to unexpected negative outcomes, the relationship between the magnitude of negative prediction errors and FRN amplitude remains a matter of debate. The present study aimed to elucidate this relationship with a new behavioural procedure that allowed subjects to predict precise reward probabilities by learning an explicit rule. Insight into the rule did not only influence subjects' choice behaviour, but also outcome-related event-related potentials. After subjects had learned the rule, the FRN amplitude difference between non-reward and reward mirrored the magnitude of the negative prediction error, i.e. it was larger for less likely negative outcomes. Source analysis linked this effect to the anterior cingulate cortex. P300 amplitude was also modulated by outcome valence and expectancy. It was larger for positive and unexpected outcomes. It remains to be clarified, however, whether the P300 reflects a positive prediction error. PMID- 18380675 TI - Right hemisphere contributions to imitation tasks. AB - Humans imitate biological movements faster than non-biological movements. The faster response has been attributed to an activation of the human mirror neuron system, which is thought to match observation and execution of actions. However, it is unclear which cortical areas are responsible for this behavioural advantage. Also, little is known about the timing of activations. Using whole head magnetoencephalography we recorded neuronal responses to single biological finger movements and non-biological dot movements while the subjects were required to perform an imitation task or an observation task, respectively. Previous imaging studies on the human mirror neurone system suggested that activation in response to biological movements would be stronger in ventral premotor, parietal and superior temporal regions. In accordance with previous studies, reaction times to biological movements were faster than those to dot movements in all subjects. The analysis of evoked magnetic fields revealed that the reaction time benefit was paralleled by stronger and earlier activation of the left temporo-occipital cortex, right superior temporal area and right ventral motor/premotor area. The activity patterns suggest that the latter areas mediate the observed behavioural advantage of biological movements and indicate a predominant contribution of the right temporo-frontal hemisphere to action observation-execution matching processes in intransitive movements, which has not been reported previously. PMID- 18380677 TI - What does "self-neglect" in older adults really cost? PMID- 18380678 TI - Discrepancies in T-score readings between patients with asymmetrical gait. PMID- 18380676 TI - Imagery or meaning? Evidence for a semantic origin of category-specific brain activity in metabolic imaging. AB - Category-specific brain activation distinguishing between semantic word types has imposed challenges on theories of semantic representations and processes. However, existing metabolic imaging data are still ambiguous about whether these category-specific activations reflect processes involved in accessing the semantic representation of the stimuli, or secondary processes such as deliberate mental imagery. Further information about the response characteristics of category-specific activation is still required. Our study for the first time investigated the differential impact of word frequency on functional magnetic resonance imaging (fMRI) responses to action-related words and visually related words, respectively. First, we corroborated previous results showing that action relatedness modulates neural responses in action-related areas, while word imageability modulates activation in object processing areas. Second, we provide novel results showing that activation negatively correlated with word frequency in the left fusiform gyrus was specific for visually related words, while in the left middle temporal gyrus word frequency effects emerged only for action-related words. Following the dominant view in the literature that effects of word frequency mainly reflect access to lexico-semantic information, we suggest that category-specific brain activation reflects distributed neuronal ensembles, which ground language and concepts in perception-action systems of the human brain. Our approach can be applied to any event-related data using single-stimulus presentation, and allows a detailed characterization of the functional role of category-specific activation patterns. PMID- 18380679 TI - Outcome of patients who present to the emergency department with an elevated international normalized ratio. PMID- 18380681 TI - Erosive pustular dermatosis of the scalp: an uncommon condition typical of elderly patients. PMID- 18380680 TI - Occurrence of suppurative parotitis in elderly people remains a bad omen. PMID- 18380682 TI - Medical complaints and psychosocial and cultural characteristics of a nationwide sample of 2,136 community-dwelling Brazilian elderly people: the Fundacao Perseu Abramo project. PMID- 18380683 TI - Atypical presentation of acute gout in an elderly patient. PMID- 18380684 TI - Botulinum toxin type A in the treatment of sialorrhea in Parkinson's disease. PMID- 18380685 TI - Lifestyle changes after oral glucose tolerance test improve glucose intolerance in community-dwelling elderly people after 1 year. PMID- 18380686 TI - Intelligence and carotid atherosclerosis in older people: cross-sectional study. PMID- 18380687 TI - Dance and movement therapeutic methods in management of dementia: a randomized, controlled study. PMID- 18380688 TI - A retrospective chart review of the tolerability and efficacy of intravenous immunoglobulin in the treatment of Alzheimer's disease. PMID- 18380689 TI - Cockcroft-Gault formula for diagnosing moderate kidney failure. PMID- 18380690 TI - Sound-alike syndromes: first report of Morvan's, and not Marfan's, syndrome. PMID- 18380691 TI - Reduced falls in the elderly: tai chi or placebo or Hawthorne effect? PMID- 18380694 TI - Validation of two different grading schemes to identify patients with asymptomatic carotid artery stenosis in general population. AB - BACKGROUND: Identification of significant asymptomatic carotid artery stenosis (ACAS) is important because of potential stroke-risk reduction offered by carotid endarterectomy. We present an external validation of two previously developed scoring schemes designed to identify patients with ACAS. METHODS: We used the data from the Cardiovascular Health Study (CHS)-a cohort study of cardiovascular risk factors, for external validation. Carotid Doppler ultrasound was performed in study participants. Two grading schemes, which used age more than 65 years, current smoking, and history of coronary artery disease and hyperlipidemia as predictors for ACAS, were validated using this dataset. RESULTS: A total of 5,449 persons (mean age 72 +/- 5 years; 42% men; and 84% white) were screened. The overall prevalence of ACAS of > or =50% was 4.2%. The prevalence of ACAS in the highest risk category was 19% in both stratification schemes. The stratification remained effective in the white sub-population (P < .001), but was not significant in the African American population (P > .05). CONCLUSION: Both schemes were effective in identifying persons with ACAS among general population aged 65 years or greater. A subset with a prevalence of ACAS of greater than 20% can be identified using these schemes making screening cost-effective among white population. PMID- 18380693 TI - In vivo imaging of neurotransmission and brain receptors in dementia. AB - Positron emission tomography (PET) and single photon emission tomography (SPECT) are the most sensitive techniques to study, in vivo, neurotransmitter systems in normal human brain and in diseases. These methods have been applied over the last years to explore the integrity of cholinergic, serotonergic, GABAergic, and dopaminergic systems in Alzheimer's disease (AD) and in other dementias. The results of these investigations have provided new insights into the neurotransmission involvement in these disorders. Moreover, these methods provide objective quantitative markers for monitoring the progression of disease and the effects of therapies. The pathophysiological and clinical relevance of results from these studies are reviewed and the potential role in early and differential diagnosis discussed. New emerging radiotracers/methods for future investigations of neurochemical processes are delineated. PMID- 18380695 TI - Transcranial ultrasound of the basal ganglia in sporadic Creutzfeldt-Jakob disease. AB - Creutzfeldt-Jakob disease (CJD) is characterized by the core symptoms of rapid progressive dementia, myoclonus, and typical periodic sharp wave complexes (PSWC) on electroencephalography (EEG). In recent years, imaging techniques have become more and more important in the diagnostic process of CJD. Magnetic resonance imaging (MRI) is very characteristic and the most accurate imaging technique to be of help in diagnosing CJD. In this study, we applied transcranial color-coded ultrasound (TCCS) in three cases of sporadic CJD. We found that all of them had in common a typical ultrasound tissue image of the lentiform nucleus, best to be described as a blurry inhomogeneous hyperechogenic signal pattern where usually the echogenicity is mostly homogeneously hypoechogenic. PMID- 18380696 TI - MRI shows dorsal lesions and spinal cord atrophy in chronic sensory neuronopathies. AB - BACKGROUND AND PURPOSE: Sensory neuronopathies (SN) represent a specific subgroup of peripheral nervous system diseases, characterized by degeneration of dorsal root ganglia (DRG) and its projections. We tried to estimate the frequency and extent of spinal cord MRI abnormalities in a group of patients with SN and correlate these with clinical and neurophysiological features. METHODS: We performed spinal cord MRI scans in 16 chronic SN patients. Images were analyzed for the presence of posterior hyperintense lesions on T2WI and cord areas at C3 level were obtained using a previously validated method. A group of 14 healthy controls with similar age and gender distribution was used for comparison. ANOVA was employed for statistical analysis. RESULTS: Posterior T2WI lesions were found in 13 out of 16 patients. Cord areas were significantly smaller in SN patients than controls (84.3 x 97.2 mm(2), P < .05). Atrophy correlated with severity of sensory ataxia and neurophysiologic abnormalities but not with duration of disease. CONCLUSIONS: These findings support volumetric spinal cord MRI as a useful tool in the assessment of chronic SN. PMID- 18380697 TI - Prenatal thrombosis of torcular Herophili with spontaneous resolution and normal outcome. AB - BACKGROUND: Prenatal thrombosis of torcular Herophili is a rare condition. It may be suspected during the routine ultrasonographic follow-up of the fetus, but MRI is necessary to establish the diagnosis. There are 7 reported cases with various results. METHODS: We report a case of prenatal torcular Herophili thrombosis. We present 2 series of fetal MRIs. The first one was performed at 21 and the second at 37 weeks' gestation. RESULTS: The second MRI revealed a total resolution of the thrombus. The product was a neurologically intact infant with normal development. CONCLUSIONS: This is the fourth reported case of fetal torcular Herophili thrombosis with normal outcome. There are no sufficient data regarding the best management of this rare condition; however, following up with fetal MRIs seems to be the most rational choice. PMID- 18380699 TI - Harvesting haemopoietic stem cells for future use: rainy days, real or imagined? PMID- 18380701 TI - Cardiac sarcoidosis. AB - Cardiac sarcoidosis can have a variety of manifestations including conduction disease, congestive heart failure, valvular heart disease, pericardial effusions, tamponade, ventricular arrhythmias and sudden cardiac death. In patients with sarcoidosis, the reported incidence of cardiac involvement ranges from 20% in US autopsy studies to nearly 60% in Japan, where it accounts for the majority of deaths as a result of sarcoidosis. Despite this, the diagnosis of cardiac sarcoidosis remains difficult and no single diagnostic test has emerged that combines a high degree of sensitivity and specificity. Recent evidence suggests that gadolinium-enhanced magnetic resonance imaging can potentially offer high sensitivity and specificity for both disease presence and activity. It may also be possible to track response to treatment of cardiac sarcoidosis with cardiac magnetic resonance imaging. Corticosteroids are the mainstay of treatment for cardiac sarcoidosis as with systemic sarcoid but at present no prospective trial has shown a survival benefit. Pharmacological treatment of heart failure should follow standard heart failure guidelines, whereas anti-arrhythmic treatment is problematic. The role of implantable cardiac defibrillators in sarcoid has not been well defined, although the risk of ventricular arrhythmias and sudden cardiac death are high. Cardiac transplantation remains an option for younger patients, although overall cardiac involvement in sarcoidosis carries a relatively poor prognosis. PMID- 18380700 TI - Consensus guidelines for 'rainy day' autologous stem cell harvests in New South Wales. AB - Autologous stem cell transplantation (ASCT) has a well-established role in the treatment of haematological malignancies. Stem cells are commonly collected following salvage chemotherapy although there may be advantages in collecting earlier in the disease course. A 'rainy day' harvest (RDH) refers to the collection of autologous haemopoietic stem cells for long-term storage. Although there are few data to support RDH, there is increasing evidence that such harvests are being carried out, creating storage pressures in stem cell laboratories across New South Wales. The Bone Marrow Transplant Network New South Wales conducted a three-staged exercise to develop consensus-based RDH guidelines. Using available evidence, guidelines were developed supporting RDH for specific patients with acute and chronic myeloid leukaemias, follicular and other lymphomas, and multiple myeloma. Physician agreement with these disease specific guidelines ranged between 58 and 100%. These consensus guidelines will improve equity of access to appropriate RDH and assist the planning of future storage requirements in New South Wales. PMID- 18380702 TI - Utility of eosinophilia as a diagnostic clue in lower abdominal pain in northern Australia: a retrospective case-control study. AB - Acute eosinophilic enteritis is a difficult diagnosis to make. Insufficient consideration of eosinophilia may commit patients to surgical treatment when medical therapy may be appropriate. The aim of the study was to determine whether the eosinophil count was considered in the diagnostic evaluation of patients presenting with acute abdominal pain who subsequently underwent appendectomy and whether eosinophilia was related to subsequent histology. The method used in the study was retrospective case-control. None of three patients with increased eosinophil counts had histologically proven appendicitis (Fisher's exact test 0.025); worm segments were seen in two patients. None of 39 patients who had histologically proven appendicitis had increased eosinophil counts. Eosinophilia may be underutilized and helminth infection may not be considered in the differential diagnosis of abdominal pain. A normal eosinophil count in the setting of clinically suspected appendicitis may make the diagnosis of eosinophilic enteritis less likely, but does not exclude it. Patients with abdominal pain and peripheral eosinophilia appear less likely to have acute appendicitis on subsequent histology; however, further study is required to validate these findings. The decision to operate remains one of clinical judgement. PMID- 18380703 TI - Warfarin-induced allergic interstitial nephritis and leucocytoclastic vasculitis. AB - Warfarin sodium has been associated with leucocytoclastic vasculitis and has once been associated with allergic interstitial nephritis. Hypersensitivity to warfarin sodium simultaneously resulting in allergic interstitial nephritis and leucocytoclastic vasculitis has not yet been previously reported. We present a 48 year-old man who was on warfarin sodium for 2 months and presented with acute renal failure and reddish purplish macules on his hypogastric regions and lower extremities bilaterally. Kidney biopsy showed allergic interstitial nephritis and punch skin biopsy showed leucocytoclastic vasculitis. Both biopsies also showed high eosinophil count, highly suggestive of a drug-induced reaction. After a negative comprehensive work-up and the absence of other recent medication changes, our patient was determined to have allergic interstitial nephritis and leucocytoclastic vasculitis secondary to warfarin sodium. PMID- 18380704 TI - Cavernous sinus thrombosis and meningitis from community-acquired methicillin resistant Staphylococcus aureus infection. AB - Septic cavernous sinus thrombosis is an uncommon clinical syndrome with a high morbidity and mortality. The commonest bacterial pathogen is Staphylococcus aureus. We describe the study of a patient with cavernous sinus thrombosis and meningitis caused by community-acquired methicillin-resistant S. aureus (CA-MRSA) infection. The isolate was genotyped as the ST93 (Queensland) clone of CA-MRSA and carried the Panton-Valentine leucocidin genes. Cure was obtained following prolonged antimicrobial therapy with vancomycin, rifampicin, cotrimoxazole and linezolid. Given the high morbidity and mortality of cavernous sinus thrombosis and the worldwide recent emergence of CA-MRSA, clinicians treating patients with this infection should consider early empirical coverage for CA-MRSA with an antimicrobial agent, such as vancomycin or linezolid, particularly in the presence of suspected facial staphylococcal skin infections. If vancomycin is used, we emphasize that high doses may be required to achieve even low levels in the cerebrospinal fluid. PMID- 18380705 TI - Statistical inference is overemphasized in cluster investigations: the case of the cluster of breast cancers at the Australian Broadcasting Corporation studios in Brisbane, Australia. AB - The aim of statistical analyses in cluster investigations is to estimate the probability that the aggregation of cases could be due to chance. As a result of several statistical problems - including the post-hoc nature of the analysis and the subjective nature of implied multiple comparisons - this cannot be carried out with any certainty. In cluster investigations, expert opinion should carry much more weight than P-values, which are exceedingly difficult to interpret. PMID- 18380707 TI - Isolated nodular pulmonary amyloidosis. PMID- 18380706 TI - Purpura fulminans in systemic lupus erythematosus. PMID- 18380708 TI - Atypical case of Herpes simplex encephalitis. PMID- 18380709 TI - Inflammatory myopathy and cancer: rare association of seminoma testes and polymyositis. PMID- 18380710 TI - Advances in multicentre ethics review. PMID- 18380711 TI - Multicentre drug trials, ethics approval and death of patients. PMID- 18380713 TI - Endothelial dysfunction in Turkish patients with non-alcoholic fatty liver disease. PMID- 18380715 TI - Hepatitis B reactivation after thalidomide. PMID- 18380716 TI - A tribute to Joe Tjandra (1957-2007). PMID- 18380717 TI - Cell system integration (CSI) trauma. PMID- 18380718 TI - An evidence-based approach: Sugarbaker protocol and pseudomyxoma peritonei of appendiceal origin. PMID- 18380719 TI - Malignant transformation in burn wound with a latent period of less than 1 year. PMID- 18380720 TI - Stapled haemorrhoidectomy, mucosectomy or anopexy: the operation is what the purse string makes it. PMID- 18380721 TI - Treatment of unresectable hepatocellular carcinoma with transarterial radioembolization: iodine-131-lipiodol. PMID- 18380722 TI - Managing deep sternal wound infections with vacuum-assisted closure. AB - Deep sternal wound infection is an uncommon but serious complication of cardiac surgery. Currently, there is no consensus on the optimal management. Vacuum assisted closure (VAC) has been increasingly used to facilitate wound healing. We aim to review the management of deep sternal wound infections using VAC dressing at our hospital. A retrospective review of consecutive cases of deep sternal wound infections was carried out. Median sternotomies were carried out in 2665 patients between July 2001 and June 2006. Thirty-one patients developed deep sternal wound infections (1.2%). In 26 of these patients, VAC dressing was used either as a stand-alone therapy or as an adjunct to late sternal reconstruction. Deep sternal wound infections were diagnosed on average 13 days from initial surgery. Of the patients treated with VAC dressing, 17 (65%) had stand-alone VAC therapy and 9 had VAC therapy followed by sternal reconstruction. The average duration of VAC dressing in the two groups were 21 and 13 days respectively. There were seven in-hospital deaths, six in the stand-alone VAC group and one death from a reconstructive patient who did not have VAC therapy. The length of hospital stay was similar in two VAC groups (37 vs 45 days). Median follow up was 17 months. No late relapse was found in the stand-alone group. In the intermediate therapy group, two patients had chronic wound sinuses and one patient had a wound collection. Vacuum-assisted closure dressing may be used both as a stand-alone and as an intermediate therapy for deep sternal wound infection. Reconstructive surgery may be avoided in a significant proportion of patients. No late relapse has been associated with VAC use. PMID- 18380723 TI - Academic surgery: does it have a future? PMID- 18380724 TI - Academic surgery in North America. PMID- 18380725 TI - Academic surgery in the United Kingdom. PMID- 18380726 TI - Academic surgery in New Zealand: dodo or phoenix? PMID- 18380727 TI - Academic surgery in Asia. PMID- 18380728 TI - Academic surgery in Papua New Guinea. PMID- 18380729 TI - Frontiers in academic surgery: the five M'S. PMID- 18380730 TI - An academic in a private hospital: a modern oxymoron. PMID- 18380731 TI - Surgical research: lost in translation? PMID- 18380732 TI - Academic surgery under threat. PMID- 18380733 TI - The college-university concept. PMID- 18380734 TI - Laparoscopic cholecystectomy in cirrhotic patients with symptomatic gallstone disease. AB - BACKGROUND: The aim of this study was to evaluate the outcome in patients with liver cirrhosis who underwent laparoscopic cholecystectomy for symptomatic gallstone disease. METHODS: Retrospective analysis of prospectively collected data of 34 patients operated between March 1998 and April 2006. RESULTS: There were 19 male and 15 female patients with a median age of 62 years. Cirrhosis aetiology was viral hepatitis in 25 patients, alcohol in 6, primary biliary cirrhosis in 2 and in 1 patient the cause was not identified. Twenty-three were classified as Child-Pugh-Turcotte stage A and 11 as Child-Pugh-Turcotte stage B. The median Model For End-Stage Liver Disease score was 12. Median operating time was 96 min. In three patients there was conversion to open cholecystectomy. Postoperatively, one patient died and six more patients had complications. Median postoperative stay was 3 days. Patients with acute cholecystitis did not have increased morbidity, but had significantly longer hospital stay. CONCLUSION: Laparoscopic cholecystectomy can be carried out with acceptable morbidity in selected patients with well-compensated Child A and B stages liver cirrhosis. Patients with evidence of significant portal hypertension and severe coagulopathy should avoid surgery. PMID- 18380735 TI - Injectable form of calcium sulphate as treatment of aneurysmal bone cysts. AB - BACKGROUND: Aneurysmal bone cysts (ABC) are a rare condition in adolescents and teenagers but may result in pain, fracture and growth abnormalities. The gold standard of open curettage carries the risk of surgical complications and still a local recurrence rate of 20-30%. Percutaneous treatment of ABC have rarely been reported and a poor response the usual outcome. This study investigated a new technique of percutaneous aspiration and injection of ABC using an aqueous solution of calcium sulphate. METHODS: A radiological diagnosis of a bone cyst was made in 15 consecutive patients and pathologically confirmed as ABC. Most had already sustained a fracture and/or had been previously unsuccessfully treated by minimally invasive techniques including embolization or methylprednisolone injection. The procedure of aspiration and injection with calcium sulphate was undertaken, and the patients were reviewed regularly both clinically and radiologically for a minimum of 2 years. RESULTS: The calcium sulphate cement was reabsorbed completely within 8 weeks. The first osseous response was periosteal new bone formation circumferentially followed by gradual opacification of the cystic cavity. All except one patient that described pain before the procedure reported complete relief of symptoms by 4 weeks. Two patients developed a local recurrence of the cyst, and one subsequently developed a pathological fracture. Two patients sustained pathological fractures through healed cysts, 12 and 22 months after the procedure, respectively. CONCLUSIONS: This new technique has shown good early clinical and radiological responses and a low complication rate in a consecutive group of patients with ABC. PMID- 18380736 TI - Treatment of unresectable hepatocellular carcinoma with radiolabelled lipiodol. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common solid organ tumours, with approximately 500,000 new cases being reported each year. It has a very high prevalence in Asia, and almost two-thirds of all cases occur in this region. The incidence of this tumour in Australia has nearly doubled in the past decade. Surgical resection is the mainstay of treatment, but only 10-30% of HCC are amenable to surgical resection at the time of diagnosis. The prognosis for patients with unresectable HCC remains dismal. Internal radiotherapy with radiolabelled lipiodol has been used with some success in treatment of HCC. Several studies have validated its usefulness in an adjuvant setting, but it also appears to have a role in treating unresectable tumours. METHODS: Twenty-two patients with proven HCC, not amenable to or having failed surgical treatment, were evaluated for radiolabelled lipiodol treatment. Of these, 10 patients were excluded after initial evaluation and 12 patients underwent treatment. Patients were followed up every 3 months with physical examination, computed tomography scan and alpha-fetoprotein levels. Overall survival, change in tumour size and alpha-fetoprotein levels were used to evaluate the efficacy of treatment. RESULTS: The median overall survival in patients undergoing lipiodol therapy was 15 months. Tumour size was stabilized in most patients and very few side-effects of the treatment were observed. CONCLUSION: This study has shown that radiolabelled lipiodol is an effective method for the treatment of unresectable locally advanced HCC. PMID- 18380737 TI - Surgical practice to reduce dialysis access insufficiency. AB - BACKGROUND: Patients who are dependant on a percutaneous central venous catheter for dialysis have an excess morbidity and mortality compared with patients with an autologous arteriovenous fistula. METHODS: In an effort to improve outcomes related to episodes of permanent access insufficiency, defined as a patient requiring a venous catheter for haemodialysis, a 12-month prospective audit of surgery carried out to establish and maintain dialysis access was carried out at our institution. RESULTS: Effective measures that reduced the period of time that patients required a venous catheter for dialysis included regular communication between dialysis staff, the surgical service and patients attending for treatment in the dialysis unit, liberal use of duplex ultrasound imaging, pursuing autologous access in the majority of patients, early intervention to correct failing vascular access before conduit thrombosis, using both traditional open surgical and endovascular solutions in establishing and maintaining vascular access, using peritoneal dialysis as a long-term or temporary alternative to haemodialysis and, in cases of arteriovenous polytetrafluoroethylene graft occlusion, early thrombectomy and mandatory revision to provide a conduit immediately available for effective dialysis. CONCLUSION: Conducting an audit of surgical practice contributed to an improvement in outcomes for dialysis dependant patients. Establishing an arteriovenous fistula in a greater proportion of cases before initiating renal replacement therapy may further address the problem of dialysis access insufficiency. PMID- 18380738 TI - Preventing postoperative haematomas in microvascular reconstruction of the head and neck: lessons learnt from 126 consecutive cases. AB - BACKGROUND: Free tissue transfer has become a safe and reliable means for repairing soft tissue and bony defects of the head and neck. Although operative success has reached 98%, the incidence of significant postoperative complications is also relatively high (32%). One common and significant complication is haematoma formation, occurring at both donor and recipient sites, and yet there are minimal published studies on its incidence, aetiology or outcome. A retrospective analysis of both donor- and recipient-site wound haematoma was carried out to identify causative factors and the effect on patient outcome. METHODS: A 5-year review of 132 consecutive microvascular free tissue transfers to head and neck defects at The Royal Melbourne Hospital, for the period February 2001 to February 2006, was conducted. RESULTS: Of 126 included cases, 27 postoperative haematomas resulted. Statistically significant associations were found for each of smoking, non-steroidal anti-inflammatory drug use and the use of corticosteroids preoperatively with the incidence of postoperative haematoma formation. Postoperative blood pressure control and the adequacy of primary tumour excision at the flap recipient site were also found to have significant associations with haematoma formation. Drain tube outputs served as accurate indicators for haematoma. CONCLUSION: There are significant reversible factors that contribute to the development of postoperative haematomas in head and neck reconstructive surgery. Preoperative modifications should, therefore, be sought. Similarly, close monitoring of patient blood pressure during the initial 24 h postoperative period by theatre and recovery staff is important, as is the adequacy of postoperative analgesia. PMID- 18380739 TI - Haemorrhoidectomy with electrocautery or ultrashears and stapled haemorrhoidopexy. AB - BACKGROUND: This study evaluated in a randomized, prospective manner the possible differences in operative time, postoperative pain and analgesic requirement, early complications, time off-work and patient satisfaction between patients undergoing haemorrhoidectomy using the electrocautery (ECH), UltraShears (ultrasonically activated scalpel; USH) or stapled haemorrhoidopexy (SH). METHODS: Patients with symptomatic third-degree or fourth-degree haemorrhoids were randomized into one of the three groups: ECH, SH (PPH stapler; Ethicon Endo Surgery, Cincinnati, OH, USA) or USH groups. The operative times and complications, if any, were recorded. An identical anaesthetic technique was used for all patients, and postoperative analgesia was also standardized according to the pain scores. Pain was evaluated preoperatively and at 8 and 24 h and 2, 7, 14 and 28 days, postoperatively, by means of a visual analogue scale. The numbers of narcotic and non-narcotic analgesics used, the time required for full return to daily activities and the incontinence scores at 4 months postoperatively were evaluated, as well as patient satisfaction. RESULTS: No significant differences with respect to age or sex existed between the three groups, each consisting of 20 patients. USH was significantly faster than both ECH and SH (P < 0.0001 for both comparisons) in operation time, and SH was faster than ECH (P < 0.0001). Comparing any two groups, the mean pain scores in the ECH group were significantly higher than in the SH and USH groups at postoperative 8 and 24 h, and day 2, but the groups reached comparable pain scores at day 7, 14 and 28. The numbers of narcotic and non-narcotic analgesics used in the ECH group were significantly higher than in the SH and USH groups (P < 0.0001 for all comparisons). The early postoperative complication rate was higher in the ECH group (25%), compared with the SH (P = 0.017) and USH (P < 0.0001) groups. Only 12 patients (60%) in the ECH group could be discharged within the first postoperative 24 h, whereas all of the patients (100%) in the SH and 17 patients (85%) in the USH groups could be treated as day cases. The time required for full return to daily activities was similar in the SH and USH groups (P = 0.841), but it was significantly longer in the ECH group (P < 0.0001 for both comparisons). The 70% patient satisfaction rate in the ECH group was significantly lower than in the SH (95%; P = 0.008) and the USH (100%, P = 0.002) groups. CONCLUSION: Our short-term results have shown that SH and ultrasonic dissector haemorrhoidectomies were superior to ECH haemorrhoidectomy, in terms of surgical utility, postoperative pain, analgesic intake, time off-work and patient satisfaction. Some advantages of USH over SH also appeared. PMID- 18380740 TI - Modified stapled haemorrhoidectomy: a suggested improved technique. AB - BACKGROUND: Longo's technique of stapled haemorrhoidopexy has revolutionized management of symptomatic haemorrhoids. However, several problems have been identified in the original Longo's technique. In particular, recurrence of haemorrhoidal tissue has been found to be significantly higher. METHODS: An improved technique with key differences to address these shortcomings is described. The modified stapled haemorrhoidectomy procedure described is carried out without the circular anal dilator and includes the placement of a low purse string of 2-3 cm. CONCLUSION: This technique is proposed as an improvement to the current stapled haemorrhoidopexy. PMID- 18380741 TI - Warm sitz bath does not reduce symptoms in posthaemorrhoidectomy period: a randomized, controlled study. AB - BACKGROUND: Warm water sitz bath is routinely recommended after anal surgery. The aim of this study was to determine the influence of sitz bath with regards to pain scores, analgesic requirements and patient satisfaction ratings after haemorrhoidectomy. METHODS: Fifty patients were randomly assigned to receive antibiotics and analgesics alone (control patients) or a twice-daily sitz bath along with identical antibiotics and analgesics (sitz bath group). Weekly pain score and patient satisfaction score were evaluated on visual analogue scores. The number of analgesic tablets consumed in each week was also assessed. RESULTS: There was no significant difference in age, gender distribution and the number of excised haemorrhoid piles between the two groups. No significant difference in postoperative mean pain score between groups (P = 0.234) was noticed. Likewise, no relevant differences in analgesic requirements between sitz bath and no sitz bath group (P = 0.435) were seen. The satisfaction score was higher in the sitz bath group when compared with the control group; however, it did not reach a statistically significant level. CONCLUSIONS: This study shows that sitz bath does not offer pain relief, wound healing or reduction in consumption of analgesics and thus there is no evidence to prescribe sitz bath in the post haemorrhoidectomy period. PMID- 18380743 TI - Splenosis presenting as multiple intra-abdominal masses mimicking malignancy. PMID- 18380744 TI - Retrorectal dermoid cyst in an adult. PMID- 18380745 TI - Medicine in small doses. PMID- 18380746 TI - Emergence of 'retropro' studies in the surgical literature. AB - BACKGROUND: The surgical published work now contains several studies that involve retrospective evaluations of prospectively collected data. These types of studies - that are referred to as 'RetroPro' studies - have the potential to be a valuable source of information about the care of surgical patients. The aim of this study was to evaluate the role of RetroPro studies in the surgical published work. METHODS: Original articles from 21 surgical journals were selected for study. Rejected from study were case reports, short communications, editorials, and surgical narratives (history, opinions and orations). Each original article was evaluated to determine the source of the study, the topic under evaluation, the sample size, the study design and the use of statistics. RESULTS: Retrospective studies accounted for 34% of the 1386 original articles. The other main components were single-group prospective studies (20%), non-clinical experimental studies (13%), RetroPro studies (10%), review articles (9%) and clinical trials (5%). Almost one-third of the RetroPro studies evaluated more than 1000 patients and 58% of them included some form of multivariable regression or an analysis of survival; that is, they attempted to model the outcome of surgical patients. CONCLUSION: RetroPro studies are now an important source of information about the outcome of surgical patients. They should be recognized as a specific form of clinical investigation. PMID- 18380747 TI - Hong Kong Museum of Medical Sciences website. PMID- 18380748 TI - Teaching autopsies. PMID- 18380749 TI - Optimizing the approach to patients with potentially resectable liver metastases from colorectal cancer. PMID- 18380750 TI - A urological alternative to the College correct patient/side/site implementation guidelines. PMID- 18380751 TI - Clostridium difficile enteritis: a cause for high ileostomy output. PMID- 18380752 TI - Gossypiboma: intramural and transmural migration causing small bowel obstruction. PMID- 18380753 TI - Acute compartment syndrome. PMID- 18380754 TI - Postoperative pseudohyperkalaemia: innocent yet dangerous. PMID- 18380755 TI - Surgical management of scimitar syndrome: an alternative approach. PMID- 18380756 TI - More time and space for congenital heart disease. PMID- 18380758 TI - Adverse event rates in congenital cardiac catheterization: a new understanding of risks. AB - We sought to characterize the frequency, severity, and attributability of adverse events (AE) in congenital cardiac catheterization and identify patient and procedural characteristics associated with AE. Risk for any AE was associated with age < 1 year (odds ratio [OR] 2.3), interventional cases (OR 2.2), increasing procedure type risk group (OR 1, 2.0, 2.5), and indicator of hemodynamic vulnerability (OR 1.6), all p < 0.001. Each of these and weight were also associated with increased risk for high severity events, except for young age. Our understanding of populations at risk for adverse outcomes in congenital cardiac catheterization cases can be enhanced through the development and refinement of variables that categorize procedure types and patient characteristics by similar risk for adverse outcomes. PMID- 18380757 TI - Early surgical morbidity and mortality in adults with congenital heart disease: the University of Michigan experience. AB - OBJECTIVES: To review early surgical outcomes in a contemporary series of adults with congenital heart disease (CHD) undergoing cardiac operations at the University of Michigan, and to investigate possible preoperative and intraoperative risk factors for morbidity and mortality. METHODS: A retrospective medical record review was performed for all patients > or =18 years of age who underwent open heart operations by a pediatric cardiothoracic surgeon at the University of Michigan Congenital Heart Center between January 1, 1998 and December 31, 2004. Records from a cohort of pediatric patients ages 1-17 years were matched to a subset of the adult patients by surgical procedure and date of operation. RESULTS: In total, 243 cardiac surgical operations were performed in 234 adult patients with CHD. Overall mortality was 4.7% (11/234). The incidence of major postoperative complications was 10% (23/234) with a 19% (45/23) minor complication rate. The most common postoperative complication was atrial arrhythmias in 10.8% (25/234). The presence of preoperative lung or liver disease, prolonged cardiopulmonary bypass and aortic cross clamp times, and postoperative elevated inotropic score and serum lactates were significant predictors of mortality in adults. There was no difference between the adult and pediatric cohorts in terms of mortality and morbidity. CONCLUSIONS: The postoperative course in adults following surgery for CHD is generally uncomplicated and early survival should be expected. Certain risk factors for increased mortality in this patient population may include preoperative presence of chronic lung or liver dysfunction, prolonged cardiopulmonary bypass and aortic cross-clamp times, and postoperative elevated inotropic score and serum lactate levels. PMID- 18380760 TI - Diagnostic role of magnetic resonance imaging in identifying aortic arch anomalies. AB - PURPOSE: The aim of this article was to assess the role of MRI in the diagnosis and management of a variety of complex aortic arch anomalies. MATERIALS AND METHODS: Imaging was performed on a 1.5T Philips Gyroscan Intera. We retrospectively reviewed all cardiac MR scans performed from November 2003 to February 2007 at our institute to identify aortic arch anomalies. Magnetic resonance imaging included three-dimensional (3D) sagittal volume images, cine images, short axis ventricular volumes, phase contrast flow of great vessels, and 3D gadolinium magnetic resonance angiogram. RESULTS: Three major categories of anomalies were identified in 47 patients: double aortic arch (n = 9), right-sided aortic arch with mirror image branching (n = 28), and aberrant right subclavian artery (n = 10). In the double aortic arch group, six patients were symptomatic, and four of them underwent a surgical division. Magnetic resonance imaging was used to plan the surgical management. In the other two groups, no patient presented with symptoms of airway or esophageal compression, and the arch abnormalities were noticed during investigation for other associated cardiac disease. However, we noticed a strong correlation with well-defined subgroups of congenital heart disease. Right-sided aortic arch was seen in patients with pulmonary atresia with ventricular septal defect (46.4%), tetralogy of Fallot (32.1%), and double outlet right ventricle with right atrial isomerism (14.2%). Seventy percent of the patients with aberrant right subclavian artery had aortic coarctation, and another case presented a complete aortic arch interruption. CONCLUSION: Magnetic resonance imaging is an important diagnostic tool in identifying anomalies of the aortic arch and its branches and can be considered the imaging technique of choice when planning surgical management, especially when there are associated cardiac anomalies. PMID- 18380759 TI - Noninvasive methods for determining pulmonary vascular function in children with pulmonary arterial hypertension: application of a mechanical oscillator model. AB - OBJECTIVE: Noninvasive diagnostics for pulmonary arterial hypertension (PAH) have traditionally sought to predict main pulmonary artery pressure from qualitative or direct quantitative measures of the flow velocity pattern obtained from spectral Doppler ultrasound examination of the main pulmonary artery. A more detailed quantification of flow velocity patterns in the systemic circuit has been obtained by parameterizing the flow trace with a simple dynamic system model. Here, we investigate such a model's utility as a noninvasive predictor of total right heart afterload and right heart function. DESIGN: Flow velocity and pressure was measured within the main pulmonary artery during right heart catheterization of patients with normal hemodynamics (19 subjects, 20 conditions) and those with PAH undergoing reactivity evaluation (34 patients, 69 conditions). Our model parameters were obtained by least-squares fitting the model velocity to the measured flow velocity. RESULTS: Five parameter means displayed significant (P < .05) differences between normotensive and hypertensive groups. The model stiffness parameter correlated to actual pulmonary vascular resistance (r = 0.4924), pulmonary vascular stiffness (r = 0.6811), pulmonary flow (r = 0.6963), and stroke work (r = 0.7017), while the model initial displacement parameter had good correlation to stiffness (r = 0.6943) and flow (r = 0.6958). CONCLUSIONS: As predictors of total right heart afterload (resistance and stiffness) and right ventricle work, the model parameters of stiffness and initial displacement offer more comprehensive measures of the disease state than previous noninvasive methods and may be useful in routine diagnostic monitoring of patients with PAH. PMID- 18380761 TI - Prevalence of migraine in adults with cyanotic congenital heart disease. AB - OBJECTIVE: There is an increased prevalence of patent foramen ovale in patients with migraine, leading to the suggestion that migraine is more common in patients with potential right-to-left shunts. The aim of this study was to investigate the prevalence of migraine in adults with large right-to-left shunts because of cyanotic congenital heart disease. DESIGN AND PATIENTS: In total, 29 cyanotic adult patients with congenital heart disease answered a questionnaire to determine the prevalence of migraine with or without aura. A total of 38 matched acyanotic patients with congenital heart disease served as controls. A subgroup of 18 acyanotic patients also underwent bubble contrast echocardiography to look for patent foramen ovale. RESULTS: Twenty (69%) of the cyanotic patients had migraine, the majority 17 (59%) having migraine with aura. Twenty-two (58%) of the 38 acyanotic patients had migraine, of whom 16 (42%) had migraine with aura. Nine (50%) of the 18 acyanotic patients who consented to an echocardiogram had patent foramen ovale. Of those with patent foramen ovale, 8 (89%) had migraine and 6 (67%) had migraine with aura. CONCLUSION: There is an increased prevalence of migraine with aura in both cyanotic and acyanotic patients with congenital heart disease. The high prevalence of migraine in acyanotic patients with congenital heart disease may be due to an increased prevalence of patent foramen ovale. PMID- 18380762 TI - Prenatal diagnosis of thoracic ectopia cordis by real-time fetal cardiac magnetic resonance imaging and by echocardiography. AB - Ectopia cordis is a rare congenital defect commonly associated with intra- and extra-cardiac anomalies. This report highlights the complimentary use of echocardiography and cardiac magnetic resonance imaging for detailed prenatal characterization of the anomaly at 23-week gestation. PMID- 18380763 TI - A pediatric case report on arrhythmogenic right ventricular dysplasia. AB - It is important for pediatric cardiologists to remember arrhythmogenic right ventricular dysplasia (ARVD) in the differential diagnosis of syncope in the pediatric population. Although the prevalence of ARVD is estimated to be relatively low and the true incidence is unknown, there are several characteristic findings that can be obtained by noninvasive modalities such as ECG, echocardiogram, and MRI that can lead to a diagnosis despite a nonspecific clinical presentation. We examined the diagnostic steps and current treatment and preventive options in a child with ARVD to highlight these features for clinicians who often evaluate children with syncope. PMID- 18380764 TI - A de novo mutation of the beta cardiac myosin heavy chain gene in an infantile restrictive cardiomyopathy. AB - Here we report the first pediatric case of restrictive cardiomyopathy secondary to a de novo mutation in the cardiac myosin heavy chain gene MYH7. The clinical course is characterized by an early onset of disease, mild hypertrophy of the left ventricle and a very short evolution to death. Because of the location of the mutation in the hinge region between the rod part and the globular head of the myosin molecule, it is possible that restrictive cardiomyopathy resulted from an impairment of flexion/extension of myosin heads during the contraction/relaxation cycle. PMID- 18380765 TI - The survival of a newborn with an interrupted aortic arch and a closed ductus arteriosus. AB - Interrupted aortic arch is a rare congenital cardiovascular anomaly with a wide spectrum of anatomical and clinical presentation. We report the case of a newborn presenting at 11 days of age with a type B-2 aortic arch interruption, a closed ductus arteriosus, and an unusual large collateral vessel arising from the ascending aorta proximal to the right carotid artery and joining an aberrant right subclavian arising from the descending aorta. PMID- 18380766 TI - Stent dilatation of a right ventricle to pulmonary artery conduit in a postoperative patient with hypoplastic left heart syndrome. AB - A 10-day-old child with hypoplastic left heart syndrome (HLHS) underwent first stage palliation for HLHS, Norwood procedure with a Sano modification, i.e., placement of a right ventricular to pulmonary artery (RV-PA) conduit. The patient developed progressively worsening systemic oxygen desaturation in the immediate postoperative period. Stenosis of the proximal RV-PA conduit was diagnosed by echocardiography. In the catheterization laboratory stent placement in the conduit was performed. This resulted in increased systemic oxygen saturation. The patient was eventually discharged from the hospital with adequate oxygen saturations. PMID- 18380767 TI - Percutaneous closure of a large PDA in a 35-year-old man with elevated pulmonary vascular resistance. AB - The presence of a large patent ductus arteriosus (PDA) may result in significant pulmonary hypertension, which may not be reversible. We present the case of a 35 year-old man with pulmonary hypertension who had successful percutaneous closure of a large PDA with an Amplatzer muscular ventricular septal defect occluder and resolution of his pulmonary hypertension. The use of prior balloon test occlusion of the PDA suggested that the procedure would be successful, despite the lack of an immediate fall in the pulmonary artery pressure. PMID- 18380768 TI - Late follow-up of patients who underwent palliation for complex congenital heart disease in childhood. AB - We report on a series of 12 patients with complex congenital heart disease who had reached adult life after surgical palliation in early childhood and who were systematically followed by clinical visits. Patients were born between 1953 and 1979 and were followed up into 2007. All patients had complex lesions with single ventricle physiology and were palliated either by banding of the pulmonary artery or by creation of systemic-pulmonary shunts. Single operations were performed in 4 cases, and 8 patients had 1 or more further palliative interventions. Late corrective surgery and heart transplantation were performed in 1 patient each. Eight survivors reached a mean age of 36 years at follow-up (range 28-48), whereas 4 patients died at a mean age of 32 years (range 22-53). All patients were in New York Heart Association classes II and III. Complications during follow-up were bacterial endocarditis (3), cerebrovascular accidents (3), arrhythmias (3), need for pacemakers (2). Deaths occurred perioperatively at transplant (1), or were caused by sudden death (2), and hypoxia related to fall in blood pressure (1). Successful pregnancies occurred in 3 patients with healthy (small) babies delivered by cesarean section in 2 of them, and vaginal delivery in 1. The course of life in these patients shows not only a wide spectrum with good quality of life in some of them but also a large number of potentially severe complications. Survival beyond 50 years is rare. PMID- 18380771 TI - The effects of warming methods on temperature, cardiac function and cytokines in plateletpheresis donors. AB - BACKGROUND AND OBJECTIVES: Plateletpheresis is the most frequent type of apheresis, with demand for these products continuously increasing. Hypothermia is a common side-effect of apheresis, which may have an effect on the donor's body functions. The aim of this study was to examine the effects of warming methods on plateletpheresis donors' temperature, cardiac function and cytokines. MATERIALS AND METHODS: Fifty plateletpheresis donors were randomly assigned to a control group (n = 25) or a warming group (n = 25), with air and blood warmers during plateletpheresis. The effects of the treatment were examined by comparing body temperature, heart rate, blood pressure, Holter EKG pattern, serum interleukin 1beta (IL-1beta), interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-alpha) concentration, the white blood count, the white blood fraction, and the platelet count at a point in time between the two groups. RESULTS: In the control group, the tympanic temperature decreased more during apheresis compared to the warming group (P = 0.014). The decrease of diastolic blood pressure was significantly greater in the control group compared to the warming group (P = 0.010). As for cardiac function, the frequency of abnormal beats was generally higher in the control group, but the difference was not significant. IL-2 and TNF-alpha decreased significantly after plateletpheresis in the control group only, while there was no change in the warming group. CONCLUSION: The decrease of temperature during plateletpheresis resulted in changes in haemodynamics and cytokines. The warming methods used in this study can prevent the decrease of temperature in donors, and may be helpful in maintaining the haemodynamic and cytokine balance. PMID- 18380772 TI - The experience of living with breast cancer. PMID- 18380773 TI - A new HLA-A*33 variant, HLA-A*3314, identified in a Korean individual. AB - The sequence of human leukocyte antigen (HLA)-A*3314 is identical to that of HLA A*330301 except for a single-nucleotide substitution at codon 49 (GCG-->GGG) resulting in an amino acid change from Ala to Gly. PMID- 18380770 TI - External quality assessment for the detection of HCV RNA, HIV RNA and HBV DNA in plasma by nucleic acid amplification technology: a novel approach. AB - BACKGROUND AND OBJECTIVES: In this EQA study a novel approach was used to assess the performance of blood centres and blood product manufacturers in detecting the possible contamination of plasma with HCV, HIV and HBV by NAT. MATERIALS AND METHODS: A panel of 12 samples, three negative and three positive for each virus, was distributed to the EQA participants. The positive samples were prepared, using the respective WHO standards, in order to obtain a viral concentration of about three times the 95% DL of the methods most commonly used by laboratories involved in blood screening by NAT. Participants were requested to test each sample of the panel on different days, possibly by different operators using their routine NAT assay. RESULTS: Overall, the participants' performance was satisfactory. In particular, 49 of the 59 participants (83%) were able to correctly identify all samples. Regarding the remaining 10 laboratories, in three cases a deviation from the laboratory's procedure that could be attributed to an operator's mistake was observed, in two cases a possible cross-contamination occurred while in the remaining five cases the failure to detect the positive samples couldn't be ascribed to any relevant deviation in the laboratory's procedure. CONCLUSIONS: The novel design of this EQA study allowed participants to verify their day by day activity as the study was carried out in the context of their routine testing. Under these conditions, it was demonstrated that, despite the high level of automation reached by NAT assays, human errors can still occur. PMID- 18380774 TI - A new HLA-A allele identified in a leukemic patient attending hematopoietic cell transplantation: A*2318. AB - Sequence-based typing procedure (SBT) procedure permitted us to identify a new human leukocyte antigen-A allele in a patient attending hematopoietic stem cell transplantation. PMID- 18380775 TI - Twenty-three novel alleles increase diversity at the HLA-C locus. AB - Twenty-three novel HLA-C alleles are described. Nine of the new alleles are single-nucleotide substitutions from their most homologous allele of which seven result in amino acid changes (Cw*0327, *0508, *0514, *0613, *0735, *0739 and *1517) and two are silent substitutions (Cw*030305 and *070107). The remaining 14 alleles (Cw*0113, *0207, *0212, *0216, *0318, *0411, *0417, *0512, *0722, *0733N, *1216, *1218, *1515 and *1607) differ from their most similar alleles by 2-4 nucleotide substitutions that result in 1-3 amino acid differences. PMID- 18380776 TI - HLA-G genotype and HLA-G expression in systemic lupus erythematosus: HLA-G as a putative susceptibility gene in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease mainly mediated by the deposit of immune complexes and defects in T lymphocytes and antigen presenting cells along with a high production of T-helper 2 cytokines. A tolerance-inducible function of nonclassical class Ib human leukocyte antigen (HLA)-G molecule in innate and adaptive cellular responses has been reported, suggesting a role in inflammatory diseases. A 14 bp sequence insertion/deletion polymorphism (rs16375) in the 3'-untranslated region of the HLA-G gene has been associated to the stability of HLA-G messenger RNA. The insertion of the 14 bp sequence seems to be associated with lower levels of soluble HLA-G (sHLA-G). The aim of this study was to evaluate the possible association of the presence of the 14 bp sequence (+14 bp) with SLE. We have HLA-G genotyped 200 SLE patients and 451 healthy control subjects (HS; Italian) and analyzed the plasma levels of sHLA G and interleukin-10 (IL-10) in a subset of SLE patients and healthy subjects (Italian and Danish). A significant increase of the +14 bp HLA-G allele was detected in the Italian SLE patients compared with HS [P = 0.003, OR 1.44 (95% CI 1.13-1.82)]. A significant increased frequency of HLA-G +14/+14 bp and a decreased frequency of HLA-G -14/-14 bp were observed in SLE patients. There median concentration of sHLA-G was significantly lower in the plasma of SLE patients compared with that in the plasma of healthy controls (P < 0.0001). Furthermore, the results confirmed higher concentrations of IL-10-positive plasma in SLE patients. These results support a potential role for HLA-G in the susceptibility of SLE. PMID- 18380777 TI - Description of two novel HLA-DPA1 alleles: DPA1*0110 and DPA1*010304. AB - DPA1*0110 was identical to DPA1*010301 except for a single-nucleotide substitution at codon 43 (TGG-->TGT) resulting in an amino acid change from Trp to Cys. DPA1*010304 was identical to DPA1*010301 except for a single silent substitution at codon 58 encoding Gly (GGC-->GGT). PMID- 18380778 TI - The study of the extended haplotypes of rare HLA-B*2730 allele using microsatellite loci. AB - The aim of the present study was to compare haplotypes of the most frequent B*27 alleles among Croatians (B*2702 and *2705) and the rare B*2730 allele. For this purpose, 37 families with members carrying human leukocyte antigen (HLA)-B27 were selected. All individuals were analysed for eight microsatellites (Msats): D6S2927, short tandem repeat - MHC class I-related gene (STR_MICA), D6S2793, D6S2811, tumor necrosis factor a (TNFa), tumor necrosis factor d (TNFd), D6S273 and D6S1014, while individuals carrying the HLA-B27 specificity were subtyped. Of 39 analysed haplotypes, 20 individuals had B*2702, 15 subjects were positive for the B*2705 allele, the B*2730 allele was found in three haplotypes from different families, while one individual carried the B*2703 allele. HLA-A3 and -DRB1*16 were shared by all three B*2730 haplotypes. The DRB1*16 allele was also observed in the majority of B*2702 haplotypes (76.5%), while HLA-A3 was, after HLA-A2, the second most frequent HLA-A specificity in B*2702 haplotypes. No such correlation was found for the B*2705 haplotypes. Msat analysis showed that B*2730 haplotypes also share the same allele at all tested Msats. The D6S2927, D6S2793, MICA and TNFd Msats were not useful in distinguishing B*2702 and B*2705 alleles because D6S2927-213bp, STR_MICA-179bp, D6S2793-206bp, D6S2811-83bp and TNFd-130bp were detected in almost all cases. Conversely, for the TNFa, D6S273 and D6S1014 loci, haplotypes carrying B*2702 and B*2730 shared a single Msat allele in the majority of cases (TNFa-113bp, D6S1014-134bp and D6S273-134bp), which was not observed for B*2705 haplotypes. In conclusion, the similarity between B*2702 and B*2730 DNA sequences as well as their sharing of the same haplotypic combinations corroborates the proposed mechanism of B*2730 evolution from B*2702 by interallelic recombination. PMID- 18380779 TI - Luminex donor-specific crossmatches. AB - In Luminex bead-based screening assays, color-coded microspheres coated with human leukocyte antigens (HLA) are used to identify both complement-binding and non-complement-binding HLA class I and II antibodies in recipient sera. Many laboratories rely on their specificity detection and use the information obtained for allocation of donor organs. A donor-specific crossmatch in the Luminex technique (LumXm) is for that reason desirable. A LumXm, in which the actual donor HLA are coated onto specific capture beads, was tested for 88 pre- and posttransplant sera of 18 recipients. The results were compared with previously published flow cytometric crossmatch (FCXm) results for the same donor-recipient combinations. All sera were also examined by Luminex single antigen (SA) tests. Class I LumXm detected 24 of 27 T-cell positive FCXm (89%) and class II 15 of 22 B-cell positive FCXm (68%). Sensitivity of LumXm for class I and II was 89% and 68% and specificity was 98% and 97%, respectively. Discrepant LumXm results were obtained in 13 sera of nine patients (15%). In general, based on SA testing, FCXm showed false-positive results for class I and LumXm gave false-negative and positive results for class II. The LumXm test was proven not to react with recipient sera containing DQ antibodies only, also DP detection was insufficient. The validity of the LumXm has been shown for class I, but its value for class II is uncertain. HLA-DR is most probably correctly identified, the validity for DQ and DP is doubtful. PMID- 18380780 TI - A HCl/alcohol formulation increased 5-aminolevulinic acid skin distribution using an ex vivo full thickness porcine skin model. AB - Topical photodynamic therapy with 5-aminolevulinic acid (5-ALA) is a newly established treatment modality for epithelial skin cancer. Skin distribution of different 20% 5-ALA formulations was investigated by detection of protoporphyrin IX (PpIX) using an ex vivo porcine skin model. Fluorescence of PpIX was first detectable at 2 h after application of a 5-ALA/HCl-alcohol solution, followed by a 5-ALA gel +/- 40% DMSO > 5-ALA lipophilic ointment > 5-ALA hydrophilic ointment at 4 h after application. Intensity of PpIX was 10-fold higher after 5-ALA/HCl alcohol application in contrast to 5-ALA hydrophilic ointment. Maxima of PpIX fluorescence were measured between 18 and 24 h after application. PpIX accumulation induced by 5-ALA within the lower parts of the epidermis was increased in order of 5-ALA hydrophilic ointment < 5-ALA lipophilic ointment < 5 ALA gel w/o DMSO < 5-ALA gel with DMSO < 5-ALA/HCl-alcohol. Illumination of skin incubated with different 5-ALA formulations led to a marked increase of apoptotic cells in the epidermis depending on the penetration depth of the 5-ALA formulations. A formulation containing short chain alcohols have been observed to increase the permeation and distribution of 5-ALA. PpIX fluorescence was detected earlier yielding to photodynamic effective amounts of PpIX in the epidermis as compared with all other 5-ALA formulations used. These data emphasizes the potential of investigating properties of new formulations using such a full thickness porcine skin model. PMID- 18380781 TI - Potential factors responsible for dryness of the dorsum of the tongue in elderly requiring care. AB - OBJECTIVE: This study explored potential factors in elderly requiring care that cause dryness of the dorsum of the tongue despite moisture retention in the sublingual mucosa. MATERIALS AND METHODS: Subjects were 224 elderly (mean age: 83.6 +/- 7.4 years) living in a nursing home and requiring individual care. Measurement of moisture of the dorsum of the tongue and sublingual mucosa was performed using a wetness tester (L-SALIVO) with a measurement time of 10 s. RESULTS: A significant difference in drying only of the dorsum of the tongue was noted depending on the degree of incapacity, level of consciousness, eating (oral ingestion or tube-feeding), and conversation. Multiple logistic regression identified the degree of incapacity in terms of eating (oral ingestion or tube feeding) as a significant factor for dryness. The odds ratio for the degree of incapacity was 3.2 while that for eating was 11.226. CONCLUSION: Dry mouth of the dorsum of the tongue in elderly requiring care was associated with the degree of incapacity, tube feeding, and level of conversation. PMID- 18380782 TI - Utilisation of dental services in a university hospital palliative and long-term care unit in Geneva. AB - OBJECTIVES: Dental care plays an important role in the multidisciplinary approach, which is used in palliative and long-term care to address the complex needs of terminally ill patients. The aim of this study was to assess the utilisation of dental services in a University Hospital Palliative and Long-term Care Unit. MATERIAL AND METHOD: Over an observation period of 13 months, structured questionnaires were filled in after each dental appointment. The survey covered three main topics: the initiation and incentive from the dental consultation, particular difficulties because of the patient's health or the hospital's organisation as well as the accomplished treatment. RESULTS: Two hundred and seventy-five questionnaires from a total of 102 patients were available for analysis. The patients' median age was 83 years (SD 10.3, range 49 101 years), 63 were female, 39 male. A majority of first appointments were initiated by a physician (n = 49 of 102), mainly because of pain (n = 62 of 275). 10.5% of the appointments were cancelled on the same day. Only one-fifth of the patients were able to reach the practice on foot. Six used a walking stick and 54 relied on a wheelchair. Eighteen patients needed to be seen in their bed. The most frequently performed procedures were extractions and removal of denture sore spots (n = 67 of 275) followed by the manufacturing of new dentures (n = 38 of 275). In more than 17% of the appointments, no particular treatment was performed. CONCLUSION: The utilisation of dental services in terminally ill and severely compromised elderly patients shown justifies a dental service in a palliative care or geriatric hospital setting. The particular dental work profile requires a practitioner with empathy and professional experience. PMID- 18380783 TI - Association between serum calcium and periodontal disease progression in non institutionalized elderly. AB - OBJECTIVE: To assess the effect of baseline serum calcium on the progression of periodontal disease in non-institutionalized elderly. BACKGROUND: Although a few studies have found some evidence of the role played by dietary calcium in periodontal disease process, there is a paucity of information pertinent to longitudinal assessment of serum calcium-periodontal relationships. MATERIAL AND METHODS: Clinical attachment levels of 266 Japanese subjects aged 70 years were recorded at baseline and annually for six consecutive years. Progression of periodontal disease (PPD) was defined as the number of teeth that showed additional attachment loss of > or =3 mm during the 6 years. The number of PPD was calculated for each subject and categorised into four levels, namely, PPD(0), PPD(1), PPD(2) and PPD(3) where the number of teeth with additional attachment loss ranged from 0, 1-10, 11-20 and >20 respectively. The levels of serum calcium, albumin, random blood sugar, immunoglobulin (IgG, IgA and IgM), gender, smoking habits, education, gingival bleeding and the number of teeth present were obtained at baseline. RESULTS: Serum calcium, IgA, smoking, gingival bleeding and teeth present were associated with PPD at p < or = 0.10 and were included in a multinomial logistic regression analysis. Serum calcium was the only variable that was significantly associated with PPD with relative risks of 100 at PPD(1) and PPD(2), respectively, and 1000 at PPD(3). CONCLUSION: Serum calcium may be considered a risk factor for periodontal disease progression in non institutionalized elderly. PMID- 18380784 TI - MSY2 and polypyrimidine tract binding protein 2 stabilize mRNAs in the mammalian testis. AB - MSY2 is a highly conserved and abundant DNA/RNA-binding protein that functions as a global stabilizer/translational suppressor of mRNAs in male germ cells. The polypyrimidine tract binding protein, PTBP2, is an RNA-binding protein that splices nuclear transcripts and stabilizes specific mRNAs in the cytoplasm. The mechanisms whereby MSY2 selects and stabilizes a large group of male germ cell mRNAs and PTBP2 stabilizes specific mRNAs such as the phosphoglycerate kinase 2 mRNA in the testis and in transfected cells will be discussed. PMID- 18380785 TI - Spermatogonial morphology, kinetics and niches in hamsters exposed to short- and long-photoperiod. AB - Previous studies have shown that under short photoperiod exposure spermatogenesis in golden hamster regresses leading to sexual inactivity. It is known that this regression is related to changes in somatic and germ cells (spermatocytes and spermatids). However, the photoperiod effects on spermatogonial biology have not been studied in detail yet. In this regard, this study was carried out to investigate the morphology, kinetics and niches of different spermatogonial types in golden hamsters under long- and short-photoperiod. Six spermatogonial generations such as type A undifferentiated (A(und)), type A differentiating (A(1), A(2), A(3)), intermediate (In) and type B spermatogonia were characterized, and were morphologically similar irrespective of the photoperiod exposure. The short photoperiod was inhibitory to A(und) spermatogonia and preleptotene but had no effect on the number of differentiating (A(1) to B) spermatogonia. In golden hamsters exposed to stimulatory-photoperiod, the interstitial components were positioned mainly in triangular areas around the seminiferous tubules and, in this situation, the A(und) spermatogonia were clearly positioned in niches (p < 0.05) in all stages studied. On the other hand, during the inhibitory-photoperiod where the seminiferous tubules have smaller diameter, the interstitial components were more homogenously distributed and the triangular areas were not clearly observed. In this case, the niches were identified only at stage VII (p < 0.05), although there was a trend of being positioned in niches area in all the stages studied. Thus, these findings suggest that the A(und) spermatogonia location in the seminiferous epithelium and the niche position are directly related to the position of the interstitial components. PMID- 18380786 TI - The frequency of undescended testis from birth to adulthood: a review. PMID- 18380787 TI - Chronic epididymitis: is epididymectomy a valid surgical treatment? AB - Chronic scrotal pain (>or=3 months) is multi-factorial in nature and difficult to treat. Epididymectomy for chronic epididymal pain is rarely performed because of perceived poor outcome. We retrospectively audited our results, when published 'cure' following testicular denervation is 97%. The records of 32 males (35 consecutive epididymectomies) were retrospectively analysed. Thirteen had previous scrotal surgery. Eight (group one) had palpable painful epididymal abnormalities on clinical examination, nine (group two) had ultrasonic abnormalities but no palpable abnormality and 15 (group three) had neither. Pain response was recorded as: cured, improved, recurred or no change/worse. The mean time to operation was 23.83 months (2-121) and follow-up was 15.57 months (1-84). There were no significant aetiological differences between groups. In group one, 87.5% were cured with the remainder improved. Sixty-seven per cent of group two had a satisfactory outcome. Of group three, 20% were cured and a further 33% improved. Prior scrotal surgery, duration of symptoms and age were not predictive of outcome (Kruskal-Wallis) in terms of pain relief. Epididymectomy for structural abnormalities had excellent results. Those with chronic pain, normal examination and ultrasound had at best, a 55% chance of improvement. This group must be counselled about the low risk of success. The identification of the optimal surgical management of this difficult problem requires a multi-national registry study. PMID- 18380788 TI - Secretory leukocyte peptidase inhibitor and lung cancer. AB - Secretory leukocyte peptidase inhibitor (SLPI) belongs to the whey acidic protein four-disulfide core family of proteins, and has antimicrobial and antiprotease functions. SLPI is produced by the epithelial cells lining the respiratory, digestive, and reproductive tracts. Gene-targeting experiments in mice indicated that one function of SLPI is to protect proepithelin from elastase cleavage in wound healing. In addition to its antiprotease function, SLPI has an anti inflammatory function through the modulation of nuclear factor-kappaB acting intracellularly, especially in macrophages. SLPI is also produced in cancer tissues, but its role in cancer is not well understood. SLPI genes are often upregulated under tumorigenic conditions. We found a negligible number of tumors in the lungs of SLPI knockout mice 20 or 40 weeks after administration of urethane, an interesting experimental model for investigating the function of SLPI in cancer. This review discusses the normal function of SLPI and its possible roles in cancer tissues. PMID- 18380789 TI - Alteration of p53-binding protein 1 expression during skin carcinogenesis: association with genomic instability. AB - Epidermal cells are the first cells to be exposed to environmental genotoxic agents such as ultraviolet and ionizing radiations, which induce DNA double strand breaks (DSB) and activate DNA damage response (DDR) to maintain genomic integrity. Defective DDR can result in genomic instability (GIN) which is considered to be a central aspect of any carcinogenic process. P53-binding protein 1 (53BP1) belongs to a family of evolutionarily conserved DDR proteins. Because 53BP1 molecules localize at the sites of DSB and rapidly form nuclear foci, the presence of 53BP1 nuclear foci can be considered as a cytological marker for endogenous DSB reflecting GIN. The levels of GIN were analyzed by immunofluorescence studies of 53BP1 in 56 skin tumors that included 20 seborrheic keratosis, eight actinic keratosis, nine Bowen's disease, nine squamous cell carcinoma, and 10 basal cell carcinoma. This study demonstrated a number of nuclear 53BP1 foci in human skin tumorigenesis, suggesting a constitutive activation of DDR in skin cancer cells. Because actinic keratosis showed a high DDR type of 53BP1 immunoreactivity, GIN seems to be induced at the precancerous stage. Furthermore, invasive cancers exhibited a high level of intense, abnormal 53BP1 nuclear staining with nuclear accumulation of p53, suggesting a disruption of DDR leading to a high level of GIN in cancer cells. The results of this study suggest that GIN has a crucial role in the progression of skin carcinogenesis. The detection of 53BP1 expression by immunofluorescence can be a useful histological marker to estimate the malignant potential of human skin tumors. PMID- 18380790 TI - Development of thermotolerance requires interaction between polymerase-beta and heat shock proteins. AB - Although heat shock proteins (HSP) are well known to contribute to thermotolerance, they only play a supporting role in the phenomenon. Recently, it has been reported that heat sensitivity depends on heat-induced DNA double-strand breaks (DSB), and that thermotolerance also depends on the suppression of DSB formation. However the critical elements involved in thermotolerance have not yet been fully identified. Heat produces DSB and leads to cell death through denaturation and dysfunction of heat-labile repair proteins such as DNA polymerase-beta (Pol beta). Here the authors show that thermotolerance was partially suppressed in Pol beta(-/-) mouse embryonic fibroblasts (MEF) when compared to the wild-type MEF, and was also suppressed in the presence of the HSP inhibitor, KNK437, in both cell lines. Moreover, the authors found that heat induced gamma H2AX was suppressed in the thermotolerant cells. These results suggest that Pol beta at least contributes to thermotolerance through its reactivation and stimulation by Hsp27 and Hsp70. In addition, it appears possible that fewer DSB were formed after a challenging heat exposure because preheat induced Hsp27 and Hsp70 can rescue or restore other, as yet unidentified, heat labile proteins besides Pol beta. The present novel findings provide strong evidence that Pol beta functions as a critical element involved in thermotolerance and exerts an important role in heat-induced DSB. PMID- 18380791 TI - Identification of SMURF1 as a possible target for 7q21.3-22.1 amplification detected in a pancreatic cancer cell line by in-house array-based comparative genomic hybridization. AB - Pancreatic cancer (PC) cell lines provide a useful starting point for the discovery and functional analysis of genes driving the genesis and progression of this lethal cancer. To increase our understanding of the gene copy number changes in pancreatic carcinomas and to identify key amplification and deletion targets, we applied genome-wide array-based comparative genomic hybridization using in house array (MCG Cancer Array-800) to 24 PC cell lines. Overall, the analyses revealed high genomic complexity, with several copy number changes detected in each line. Homozygous deletions (log(2)ratio < -2) of eight genes (clones) were seen in 14 of the 24 cell lines, whereas high-level amplifications (log(2)ratio > 2) of 10 genes (clones) were detected in seven lines. Among them, we focused on high-level amplification at 7q22.1, because target genes for this alteration remain unknown. Through precise mapping of the altered region by fluorescence in situ hybridization, determination of the expression status of genes located within those regions, and functional analysis using knockdown of the gene expression or the ectopic overexpression approach in PC cell lines, as well as immunohistochemical analyses of candidates in primary tumors of PC, we successfully identified SMURF1 as having the greatest potential as a 7q21.3-22.1 amplification target. SMURF1 may work as a growth-promoting gene in PC through overexpression and might be a good candidate as a therapeutic target. Our results suggest that array-based comparative genomic hybridization analysis combined with further genetic and functional examinations is a useful approach for identifying novel tumor-associated genes involved in the pathogenesis of this lethal disease. PMID- 18380792 TI - Improvement of the breaking strength of wound by combined treatment with recombinant human G-CSF, recombinant human M-CSF, and a TGF-beta1 receptor kinase inhibitor in rat skin. AB - Effective doses of ionizing radiation during preoperative radiotherapy occasionally cause wound complications after subsequent surgery. The authors attempted to accelerate radiation-impaired wound healing in animal models. Recombinant human granulocyte colony-stimulating factor (rhG-CSF), recombinant human macrophage colony-stimulating factor (rhM-CSF), and an inhibitor of transforming growth factor (TGF)-beta1 receptor kinase, SB431542, were injected s.c. into a full-thickness incisional wound site in the dorsal skin of rats after local irradiation of X-ray (30 Gy). Wound healing of irradiated skin was assessed using the breaking strength of the wound and histological analyses. The impaired wound healing in irradiated skin was found to be associated with impaired mobilization of bone marrow-derived cells and enhanced expression of TGF-beta1 mRNA. The breaking strength of the wound in the irradiated skin was approximately one-eighth of that in the non-irradiated skin; however, following combined treatment with the above three compounds the breaking strength increased to approximately one-half of that in the non-irradiated skin. Histological analysis of the wounded skin revealed an increase in formation of collagen fibers and the panniculus carnosus following the combined treatment. Moreover, the increased breaking strength was associated with an increase in a subpopulation of fibrocytes (collagen I/ED1 double positive cells). These findings suggested that a combined treatment with rhG-CSF, rhM-CSF, and SB431542 is promising as a means of improving radiation-impaired wound healing. PMID- 18380794 TI - Induction of multinucleated cells and apoptosis in the PC-3 prostate cancer cell line by low concentrations of polyethylene glycol 1000. AB - Polyethylene glycol (PEG) has been reported to inhibit the development of colonic lesions in carcinogen-treated rats when administered orally. However, the precise mechanism for the chemopreventive activity of PEG remains largely elusive. Based on a characteristic feature of PEG as a 'fusogen', we investigated its potential as a chemotherapeutic agent through the induction of multinucleated cell formation and apoptosis induction in PC-3 prostate cancer cells. When PC-3 cells were treated with 0.5 and 1.0% PEG 1000, multinucleated cells were induced at a frequency of 8.4 and 13%, respectively, 36 h after PEG treatment under high cell density (1 x 10(6) cells in 100 microL PEG solution) in vitro. Although abnormality of cell cycle progression was not evident in PEG-treated PC-3 cells, multinucleated cells substantially disappeared at around 38 h due to apoptosis. In contrast, no apparent growth suppression was observed when PC-3 cells were exposed to up to 1.0% PEG at a much lower cell density, namely under ordinary culture conditions. Furthermore, injection of 0.5% PEG solution in vivo into PC-3 xenografts implanted in BALB/c-nu/nu male mice significantly suppressed tumor growth compared to phosphate-buffered saline injection. Multinucleated TdT mediated dUTP-biotin nick end-labeling (TUNEL)-positive cells were observed inside the PEG-injected tumors. PEG was here demonstrated to have anticell proliferation and antitumor effects via induction of apoptosis, possibly by cell fusion. PEG injection therapy could therefore be adopted as an alternative chemotherapeutic strategy for localized prostate cancers, including those that become refractory to androgen-deprivation therapy. PMID- 18380793 TI - CYP2A6 and the plasma level of 5-chloro-2, 4-dihydroxypyridine are determinants of the pharmacokinetic variability of tegafur and 5-fluorouracil, respectively, in Japanese patients with cancer given S-1. AB - S-1 is an oral anticancer agent composed of tegafur (FT), 5-chloro-2,4 dihydroxypyridine (CDHP), and potassium oxonate. CDHP is added to prevent degradation of 5-fluorouracil (5-FU) by inhibiting dihydropyrimidine dehydrogenase. CYP2A6 is involved in the biotransformation of FT to 5-FU. Thus, we prospectively analyzed the effects of the CYP2A6 genotype, plasma level of CDHP, and patient characteristics on the pharmacokinetic (PK) variability of FT and 5-FU. Fifty-four Japanese patients with metastatic or recurrent cancers who received S-1 were enrolled. The CYP2A6 polymorphisms (*4A, *7, and *9) with deficient or reduced activity were analyzed. All subjects were classified into three groups according to their CYP2A6 genotype: wild type (*1/*1), one-variant allele (*1/any), or two-variant alleles (combination other than *1). The PK of FT, 5-FU, and CDHP were measured on day 1 of treatment. Multivariate regression analysis revealed that oral clearance of FT was associated with the CYP2A6 genotype (analysis of variance [ANOVA], P = 0.000838). The oral clearance of FT seen in patients with the two-variant alleles was significantly lower than those in wild type and the one-variant allele (95% confidence intervals 0.75-2.41 and 0.41-1.82, respectively; Tukey-Kramer test). The area under the time concentration curve (AUC) of 5-FU was significantly correlated with the AUC of CDHP (ANOVA, P = 0.00126). The AUC of 5-FU and CDHP were inversely correlated with creatinine clearance (ANOVA, P = 0.0164 and P = 0.000762, respectively). Although the CYP2A6 variants are the cause of the PK variability of FT, the AUC of CDHP affected by renal function is the key determinant of the variability in the PK of 5-FU. PMID- 18380795 TI - Downregulation of Tie2 gene by a novel antitumor sulfolipid, 3'-sulfoquinovosyl 1'-monoacylglycerol, targeting angiogenesis. AB - We previously reported that 3'-sulfoquinovosyl-1'-monoacylglycerol (SQMG) was effective in suppressing the growth of solid tumors due to hemorrhagic necrosis in vivo. In the present study, we investigated the antiangiogenic effect of SQMG. In vivo assessment of antitumor assays showed that some tumor cell lines, but not others, were sensitive to SQMG. Microscopic study suggested that in SQMG sensitive tumors, but not SQMG-resistant tumors, angiogenesis was reduced. We next investigated gene expression relating to angiogenesis in tumor tissues by quantitative real-time polymerase chain reaction. Consequently, although vascular endothelial growth factor gene expression was not detected with significant differences among the cases, significant downregulation of Tie2 gene expression was observed in all SQMG-sensitive tumors as compared with controls, but not in SQMG-resistant tumors. These data suggested that the antitumor effects of SQMG could be attributed to antiangiogenic effects, possibly via the downregulation of Tie2 gene expression in SQMG-sensitive tumors. PMID- 18380796 TI - Evaluation of the one-minute exercise test to detect peripheral arterial disease. AB - BACKGROUND: Asymptomatic peripheral arterial disease (PAD) is common amongst the elderly and is a risk factor for cardiovascular morbidity and mortality. PAD can be assessed by non-invasive tests such as the ankle/brachial pressure index (ABPI) at rest and Doppler flow velocity (DFV) scanning, but these tests may underestimate the prevalence of PAD. The aim of this study was to estimate the added value, for the detection of PAD, of the one-minute exercise test, defined as positive if the drop of the ankle systolic pressure was more than 30 mmHg. We also investigated whether the combination of the ABPI at rest and the one-minute exercise test could replace DFV scanning. MATERIALS AND METHODS: We studied this in a random sample (n = 631) of a 50- to 75-year-old population. RESULTS: Of these subjects 11% (66/631) had an abnormal ABPI (< 0.9) and 16% (102/631) had an abnormal DFV curve. Of this sample 72% of the subjects performed a one-minute exercise test. Of all subjects 6% (27/451) had an abnormal ABPI (< 0.9) and 12% (54/451) had an abnormal DFV curve. The one-minute exercise test revealed seven cases of PAD (beyond the 67 already identified) which were not detected by an abnormal ABPI at rest and/or DFV scanning. As a result the prevalence of PAD increased by 2%. All patients with an aortoiliac or femoropopliteal obstruction had an ABPI at rest < 0.9. The sensitivity of the combination of the ABPI at rest and the one-minute exercise test to detect abnormal DFV curves was low for crural obstructions. CONCLUSION: The one-minute exercise test slightly improves the detection of peripheral arterial disease in the general population. PMID- 18380797 TI - Inhibition of inducible nitric oxide synthase activity prevents liver recovery in rat thioacetamide-induced fibrosis reversal. AB - BACKGROUND: Stimulation of nitric oxide (NO) synthesis similar to the application of NO donors could be of benefit in liver fibrosis. Many authors believe that activation of NO synthesis by pharmacological agents is promising in the treatment of liver fibrosis. However, there is considerable controversy in understanding the role of NO in fibrogenesis and fibrolysis. The aims of our study were to evaluate the effects of L-arginine, as an NO metabolic precursor, and those of NO synthase (NOS) inhibitors, L-nitroarginine methyl ester (L-NAME) and aminoguanidine (AG) in rats with thioacetamide (TAA)-induced liver fibrosis reversal. MATERIALS AND METHODS: Male Wistar rats, 230-240 g, received TAA (200 mg kg(-1), intraperitoneally) twice a week for 3 months. Liver resolution was simulated by withdrawal of TAA administration. Thereafter the animals were subdivided into five groups and treated by intragastric intubation with: L arginine (100 and 300 mg kg(-1)); L-NAME as an inhibitor of both constitutively expressed NOS (eNOS) and inducible NOS (iNOS) (20 mg kg(-1)), AG as a specific inhibitor of iNOS (100 mg kg(-1)) or placebo. The severity of liver fibrosis was assessed by morphometric evaluation of liver slides stained with Azan-Mallory, hydroxyproline (Hyp) determination and mRNA steady state levels of collagen I, transforming growth factor (TGF)-beta1, metalloproteinases (MMP)-13, -14, tissue inhibitor of MMP (TIMP)-1 and plasminogen activator inhibitor (PAI)-1 were quantified by real time PCR. The activities of serum marker enzyme, alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase, were measured. RESULTS: TAA treatment during 3 months induced micronodular liver fibrosis with a pronounced deposition of collagen fibres. L-Arginine did not affect this deposition nor did it affect both relative and total liver hydroxyproline content. Both NOS inhibitors significantly increased the square of the liver connective tissue stained by Azan-Mallory and the above parameters characterizing liver hydroxyproline content. Both NOS inhibitors up-regulated procollagen alpha1 (I), MMP-13, TIMP-1 and PAI-1 mRNA expression. The AG effects were more pronounced. than those of L-NAME. AG treatment also increased mRNA expression of TGF-beta1 and PAI-1. CONCLUSIONS: Both NOS inhibitors developed a clear pro-fibrotic effect in the liver. Aminoguanidine was more fibrotic than L NAME. Our data suggest a significant anti-fibrotic role for iNOS rather than for eNOS. L-Arginine did not show any anti-fibrotic properties in the TAA-model used. PMID- 18380798 TI - Sex hormone sensitivity of matrix metalloproteinases 6 and 9. PMID- 18380799 TI - Abstracts of the Clinical Vascular Biology Congress, 7-11 May 2008, Antalya, Turkey. PMID- 18380800 TI - Mortality risks associated with Barrett's oesophagus. PMID- 18380802 TI - Protective effect of nasal immunization of influenza virus hemagglutinin with recombinant cholera toxin B subunit as a mucosal adjuvant in mice. AB - To develop an efficient nasal influenza vaccine, influenza A and B virus HA with rCTB as a mucosal adjuvant were administered to mice intranasally. Serum anti-HA IgG and IgA antibody responses for both HA vaccines were significantly increased in the presence of rCTB. Higher HI and neutralizing antibody titers and higher mucosal IgA antibody responses in the respiratory tract were detected when rCTB was added than without rCTB. When mice were immunized with HA vaccine with or without rCTB and challenged by intranasal administration of mouse-adapted pathogenic influenza A virus, all mice immunized with HA plus rCTB survived for seven days without any inflammatory changes in the lungs, while not all the mice immunized with HA without rCTB survived, and all of them had lung consolidations. These results demonstrate that intranasal co-administration of rCTB as a mucosal adjuvant with influenza virus HA is necessary not only for the induction of systemic and mucosal HA antibodies, but also for the protection of mice from morbidity and mortality resulting from virus infection. PMID- 18380804 TI - Hsa, an adhesin of Streptococcus gordonii DL1, binds to alpha2-3-linked sialic acid on glycophorin A of the erythrocyte membrane. AB - Bacterial recognition of host sialic acid-containing receptors plays an important role in microbial colonization of the human oral cavity. The aggregation of human platelets by Streptococcus gordonii DL1 is implicated in the pathogenesis of infective endocarditis. In addition, we consider that hemagglutination of this organism may act as an additive factor to increase the severity of this disease. We previously reported that this interaction requires the bacterial expression of a 203-kDa protein (Hsa), which has sialic acid-binding activity. In the present study, we confirmed that erythrocyte surface sialoglycoproteins are the receptors for Hsa. We examined the effects of proteinase K, chymotrypsin, phospholipase C, and alpha(2-3) or alpha(2-3, 6, 8) neuraminidase on hemagglutination activity and found that the interaction occurs between Hsa and alpha2-3-linked sialic acid containing proteins of erythrocytes. We expressed recombinant NR2, which is the putative binding domain of Hsa, fused with GST in Escherichia coli BL21. Dot-blot analysis demonstrated that GST-HsaNR2 binds both glycophorin A (GPA) and band 3. Moreover, GPA and a small amount of band 3 were detected by GST pull-down assays. These findings indicate that S. gordonii Hsa specifically binds to GPA and band 3, alpha2-3-linked sialic acid membrane glycoproteins. PMID- 18380803 TI - New serotype of mutans streptococci isolated from pig oral cavity. AB - Gram-positive streptococcal mutans-like strains, but with clearly different colony formation than S. orisuis on Mitis Salivarius agar, were isolated from the pig oral cavity and identified by 16S rRNA sequencing, G+C content, DNA-DNA homology and extensive biochemical and serological testing. The phenotypic data showed that the strains were similar to S. orisuis except for susceptibility to bacitracin. DNA-DNA homology between the isolates and S. orisuis was 72~81%. However, serological data showed that they have a different sero-specific antigen from S. orisuis and other mutans streptococci. A new serotype, designated p, strains are classified in a serovar of S. orisuis, one of mutans streptococci. PMID- 18380805 TI - Human immunodeficiency virus type-1 vulnerates nascent neuronal cells. AB - Macrophages or microglial cells are the major target cells for HIV-1 infection in the brain. The infected cells release neurotoxic factors that may cause severe neuronal cell damage, especially in the basal ganglia and hippocampus. In this study, we used rat OHC to examine the region-specific neuronal cell damage caused by HIV-1-infected macrophages. When OHC was cocultured with HIV-1-infected MDM, we found that neuronal cells at the GCL of the DG were preferentially killed via apoptosis, and that projection of MF from GCL to PCL of the CA3 region was severely disturbed. We marked precursor cells around the DG region by using an EGFP-expressing retrovirus vector and found that these cells lost the ability to differentiate into neurons when exposed to HIV-1-infected MDM. In the DG, new neurons are normally incorporated into GCL or PCL, while in the presence of HIV-1 infected MDM, mature neurons failed to be incorporated into those layers. These data indicate that the neurotoxic factor(s) released from HIV-1-infected macrophages impede(s) neuronal cell repair in brain tissue. This suggests that DG is the region of the hippocampus most vulnerable to neuronal damage caused by HIV 1 infection, and that its selective vulnerability is most likely due to the highly active neurogenesis that takes place in this region. PMID- 18380806 TI - Generation and characterization of human monoclonal antibodies to G5, a linear neutralization epitope on glycoprotein of rabies virus, by phage display technology. AB - The aim of the present study was to discover distinct human MAbs to RV with high neutralizing potency and a broad neutralization spectrum. A phage display technology was used to produce human scFv to G5, a conserved linear neutralization epitope on Gp of RV. A phage display scFv library with 6 x 10(7) members was constructed and the phage-scFv with 'antigen-binding' activities were selected with synthetic peptide G5-24. The obtained scFv genes were cloned into pET22b(+)/BL21(DE3) and from this we prepared purified scFv fragments. The assay of the specificity characteristics and neutralization capacity showed that two distinct clones with new human immunoglobulin V genes can recognize G5 specifically as well as neutralize different RV strains. They have potential for inclusion in an antibodies combination aimed for use in rabies PEP. PMID- 18380807 TI - Disruption of the association of 73 kDa heat shock cognate protein with transporters associated with antigen processing (TAP) decreases TAP-dependent translocation of antigenic peptides into the endoplasmic reticulum. AB - Major histocompatibility complex class I-bound antigenic peptides generated in the cytosol are translocated into the ER by TAP. In the present study, the physical association of HSC73 with TAP in human lymphoblastoid T1 cells was demonstrated. The dissociation was induced in the presence of 10 mM ATP, indicating that the ADP-binding form of HSC73 might be associated with TAP. We found that HSC73-binding immunosuppressant, MeDSG disrupted the HSC73-TAP association, whereas it did not affect the binding of HSC73 to a substrate protein. MHC class I expression on the cell surface was also downregulated. Then, the effect of MeDSG on the TAP-mediated ER translocation was examined using two homologous model peptides, NGT-Bw4 and NGT-Bw6, which had distinct binding affinity to HSC73. Although high-affinity peptide NGT-Bw4 was translocated by TAP, low-affinity peptide NGT-Bw6 was not. The TAP-dependent translocation of NGT Bw4 was abolished in the presence of MeDSG. Decreased presentation on the cell surface was shown for the human leukocyte antigen (HLA)-A31-restricted natural antigenic peptide F4.2, which had high affinity to HSC73, in the presence of MeDSG. It was indicated that disruption of the HSC73-TAP association resulted in inhibition of TAP-dependent translocation of HSC73-bound peptides. Our findings highlighted an important role of HSC73 for feeding antigenic peptides to TAP, and suggested a possibility that a synthetic polyamine might inhibit the function of HSC73, thereby suppressing MHC class I-restricted presentation of HSC73-bound antigenic peptides. PMID- 18380808 TI - Attenuated Listeria infection activates natural killer cell cytotoxicity to regress melanoma growth in vivo. AB - Listeria monocytogenes infection induces various types of immune responses. The Lm-induced immunity not only protects the hosts against Lm infection but also has a therapeutic effect on other diseases such as tumors and infectious diseases. In the present study, we sought to identify the cells and molecules that are primarily responsible for the Lm-induced antitumor immune response. We investigated the mechanism of the antitumor immune response induced by Lm infection using melanoma cells and various types of gene-manipulated mice and B16F10 melanoma cells. Melanoma cells were implanted into mice intrasplenically or intraperitoneally. Lm infection of mice remarkably suppressed the growth of transplanted melanoma. The suppression of melanoma growth was due to the augmented NK cytotoxicity. The Lm-induced NK activation against melanoma was type I interferon- and signal transducer and activator of transcription (STAT)1 dependent but independent of IL-12 and IFN-gamma. In contrast to avirulent Listeria innocua and hly(-) Lm failed to induce NK activation, a mutant Lm strain with minimal hemolytic activity and with normal accessibility to cytoplasm induced NK activation. We demonstrated that the attenuated Lm entrance into the cytoplasm induces the production of type I IFN followed by the activation of NK cells, which is essential for the Lm-induced antitumor response. PMID- 18380809 TI - Co-infection of respiratory bacterium with severe acute respiratory syndrome coronavirus induces an exacerbated pneumonia in mice. AB - SARS-CoV grows in a variety of tissues that express its receptor, although the mechanism for high replication in the lungs and severe respiratory illness is not well understood. We recently showed that elastase enhances SARS-CoV infection in cultured cells, which suggests that SARS development may be due to elastase mediated, enhanced SARS-CoV infection in the lungs. To explore this possibility, we examined whether co-infection of mice with SARS-CoV and Pp, a low-pathogenic bacterium which elicits elastase production in the lungs, induces exacerbation of pneumonia. Mice co-infected with SARS-CoV and Pp developed severe respiratory disease with extensive weight loss, resulting in a 33~90% mortality rate. Mice with exacerbated pneumonia showed enhanced virus infection in the lungs and histopathological lesions similar to those found in human SARS cases. Intranasal administration of LPS, another elastase inducer, showed an effect similar to that of Pp infection. Thus, this study shows that exacerbated pneumonia in mice results from co-infection with SARS-CoV and a respiratory bacterium that induces elastase production in the lungs, suggesting a possible role for elastase in the exacerbation of pneumonia. PMID- 18380810 TI - The Bordetella type III secretion system: its application to vaccine development. AB - B. pertussis is a causative agent of whooping cough (pertussis) in humans. Despite wide-scale vaccination in many countries, there is serious concern about pertussis as a re-emerging disease. Re-emergence of pertussis may be explained by several factors: the short duration of protection by the currently available acellular pertussis vaccine, an increase in asymptomatic adult carriers and expansion of strains with certain antigenic variations which are not covered by currently available vaccines. To develop safer and more efficacious vaccines which confer more prolonged protection, researchers are focusing on identification and characterization of new virulence factors. One candidate for protective antigens is the type III secretion system and its secreted proteins. PMID- 18380811 TI - Hematuria: definition and screening test methods. PMID- 18380812 TI - Economic evaluation of prostate cancer screening with prostate-specific antigen. AB - Economic issues cannot be ignored in conducting prostate cancer screening using prostate-specific antigen (PSA). Through an electronic search, we reviewed five descriptive cost studies and nine cost-effectiveness/cost-utility analyses concerning PSA screening. Most of the existing evidence was based on mathematical model analysis and the results are enormously disparate. The cost per quality adjusted life years (QALY) gained was estimated to be $US 63.37 to $68.32, and $8400 to $23 100, respectively, or was dominated by no screening. Economic studies evaluating PSA screening are still far from sufficient. Urologists, epidemiologists and health economists must jointly conduct further studies on not only mortality but also quality of life assessment and economic evaluation, using randomized clinical trials, for a strict evaluation of the actual efficacy of PSA screening. At present, patients should be thoroughly informed of the limitations of PSA screening and, in consultation with urological specialists, make the personal decision of whether to receive it. PMID- 18380813 TI - Prostate stem cells: the niche and cell markers. AB - Prostate cancer and benign prostatic hyperplasia are common diseases in elderly men worldwide. Identifying the prostate stem cell is an important tool to investigate the mechanism of these prostatic diseases. Although the prostate stem cell has not yet been detected, progress has been made. The 'niche' or place in which the prostate stem cell resides is thought to be located in the proximal region of the murine prostate, near the urethra. Several candidate prostate stem cell markers are currently under investigation. In this review, we summarize the historical approaches and recent evidence regarding the niche and prostate specific stem cell markers. PMID- 18380814 TI - Laparoscopic adrenalectomy for malignant tumors. AB - OBJECTIVES: The treatment of malignant adrenal tumors using laparoscopic surgery remains controversial. We thus compared the perioperative outcome of the laparoscopic adrenalectomy for the treatment of malignant tumors with the outcome for benign tumors. We also evaluated the oncological outcome of the laparoscopic adrenalectomy for a malignancy. METHODS: Since 1999 a total of nine laparoscopic adrenalectomies for a malignancy have been performed in nine patients. The median adrenal tumor size was 3 cm. The laparoscopic approach was transperitoneal in all cases. Seven patients had no evidence of a systemic metastatic disease, whereas two patients with a metastatic renal cell carcinoma had systemic metastatic disease at the time of the operation. RESULTS: The median operation time was 165 min and the estimated blood loss was 75 mL in the laparoscopic adrenalectomy for a malignancy. There was no significant difference between laparoscopic adrenalectomy for malignant and benign tumors. Regarding the oncological outcome, seven of the nine patients, including the two palliative cases, treated with a laparoscopic adrenalectomy for a malignancy were alive at a median follow-up of 20 months. One patient died of other causes. CONCLUSIONS: Our results clearly indicate that a laparoscopic adrenalectomy for the treatment of a metastatic adrenal malignancy can be performed with an acceptable outcome as a minimally invasive method in carefully selected patients. PMID- 18380815 TI - How do symptoms have an impact on the prognosis of renal cell carcinoma? AB - AIM: Symptomatic renal cell carcinoma (RCC) is well known to have a characteristic behavior. We therefore evaluated the impact of systemic symptoms on the prognosis of RCC. METHODS: Patard's criteria were used to classify symptoms before operation into three groups defined as: S1 (incidental tumor), S2 (localized symptoms) and S3 (systemic symptoms). Selected clinicopathological factors including gender, maximum tumor diameter, clinical stage, hemoglobin, C reactive protein (CRP) and immunosuppressive acidic protein, nuclear grade and venous invasion were measured preoperatively in 252 patients. To determine impacts of them on the prognosis of RCC, we compared quantitative results using Cox multivariate analysis. RESULTS: The cancer-specific five-year survival rates were 93.1%, 71.0%, and 20.2% for S1 (144 patients), S2 (80 patients) and S3 (28 patients), respectively (P < 0.0001). By the univariate analysis, S2 and S3 were significant prognostic factors (risk ratio 4.5, P = 0.0003, risk ratio 19.15, P < 0.0001, respectively). By the multivariate analysis limited to preoperative clinical characteristics, S3 and CRP were independent factors (risk ratio 7.05, P = 0.0006, risk ratio 3.53, P = 0.0052, respectively). When pathological factors as well as preoperative clinical features were included on multivariate analysis, S3 and CRP remained to be independent predictive factors (risk ratio 6.01, P = 0.0031, risk ratio 2.64, P = 0.0040, respectively). Among pathological factors, only nuclear grade was a significant prognostic factor (risk ratio 2.92, P = 0.013). CONCLUSION: The presence of systemic symptoms is an independent prognostic factor along with nuclear grade and CRP. PMID- 18380816 TI - Evaluation of the association of current cigarette smoking and outcome for patients with clear cell renal cell carcinoma. AB - OBJECTIVES: Cigarette smoking is a well known risk factor for the development of renal cell carcinoma (RCC); however, its association with tumor aggressiveness and patient outcome remains in question. Herein, we test the hypothesis that cigarette smoking is associated with a more aggressive phenotype and poorer outcome among patients with RCC. METHODS: We examined data on 2242 patients treated with radical nephrectomy or nephron-sparing surgery for unilateral, sporadic, clear cell RCC at Mayo Clinic Rochester between 1970 and 2002. Associations of self-reported smoking status with death from RCC were assessed using Cox proportional hazards regression models summarized with hazard ratios (HR) and 95% confidence intervals (CI). RESULTS: While former cigarette smoking was not associated with an increased risk of RCC death, current cigarette smokers were 31% more likely to die from RCC compared with non-smokers on a hazard ratio scale (HR 1.31; 95% CI 1.09-1.58; P = 0.004). Interestingly, current smokers were more likely to present with advanced disease (i.e. later TNM stage) compared with both former and never smokers. After adjustment for TNM stage group and tumor grade, there was no longer a statistically significant increase in the risk of death from RCC for current cigarette smokers (HR 0.99; 95% CI 0.82-1.19; P = 0.875). CONCLUSIONS: Patients who report current smoking at time of surgery are at increased risk of RCC death; however, this association is attenuated after adjustment for standard pathological indices and is therefore of little prognostic value. Nevertheless, the association of current smoking with more advanced disease at presentation (e.g. metastatic spread) warrants further investigation. PMID- 18380817 TI - Clinical outcomes of bacillus Calmette-Guerin instillation therapy for carcinoma in situ of urinary bladder. AB - OBJECTIVES: We analyzed the clinical outcomes of instillation therapy with Bacillus Calmette-Guerin (BCG) to treat carcinoma in situ (CIS) and searched for prognostic factors that could predict disease progression. METHODS: Between January 1995 and January 2001, 185 patients (male, 155; female, 30) diagnosed with bladder CIS underwent weekly BCG instillations (80 mg of Tokyo 172 strain) for eight weeks. Primary, concomitant, and secondary CIS was found in 62 (33.5%), 60 (32.4%) and 63 (34.1%), patients, respectively. Seventy-five (40.5%) and 64 (34.6%) patients had limited and extensive CIS, respectively. The median follow up period was 37.5 months (range 4-95 months). RESULTS: The overall complete response rate was 86.5%. The five-year progression-free survival rate was 78.5%. Several factors, such as age (<60 or >or=60 years), gender, previous transurethral resection, type of CIS, and CIS extension (three or more positive sites out of four to six biopsy sites was defined as extensive), were examined by multivariate analysis to predict progression. The extension of CIS was the only independent prognostic factor. The five-year recurrence-free rate of complete responders (n = 160) was 66.0%. Radical cystectomy was performed in 10 patients (6.3%) during follow up incomplete responders, of whom seven had invasive bladder cancer. Extravesical involvement was identified in 30 patients (16.2%) among whom, 21 (11.3%) had upper urinary tract recurrence and nine (4.9%) had prostatic involvement. CONCLUSION: Therapy with BCG is effective against CIS, the extent of which might be a prognostic factor. Disease progression including extravesical involvement should be carefully monitored over the long-term after BCG therapy. PMID- 18380818 TI - Prognostic significance of adjuvant cisplatin-based combination chemotherapy following radical cystectomy in patients with invasive bladder cancer. AB - OBJECTIVES: The objectives of this study were to retrospectively examine the outcomes of adjuvant cisplatin-based combination chemotherapy following radical cystectomy in patients with invasive bladder cancer in order to identify parameters predicting the prognosis of these patients. METHODS: This study included a total of 51 patients with muscle invasive bladder cancer who underwent radical cystectomy between January 1995 and December 2004, and subsequently received at least two cycles of either MVAC (methotrexate, vinblastine, doxorubicin and cisplatin) or MVP-CAB (methotrexate, doxorubicin, vincristine, cyclophosphamide, bleomycin and cisplatin) chemotherapy in an adjuvant setting because of the presence of diseases corresponding to pT3/4 and/or pN1/2 without clinical evidence of metastasis. The significance of several clinicopathological factors as predictors of disease recurrence and cancer-specific death was evaluated using univariate and multivariate analyses. RESULTS: During the observation period of this study (median, 26.5 months), disease-recurrence and cancer-specific death were observed in 21 and 16 patients, respectively. The 5 year recurrence-free and cancer-specific survival rates were 48.6% and 54.1%, respectively. Among several factors examined in this series, lymph node metastasis and concomitant carcinoma in situ (CIS) were identified as significant predictors of both disease-recurrence and cancer-specific death, and these were also independently associated with disease-recurrence and cancer-specific death. CONCLUSIONS: Adjuvant cisplatin-based combination chemotherapy for patients with extravesically extended bladder cancer following radical cystectomy resulted in comparatively unsatisfactory outcome concerning cancer-control; accordingly, it would be necessary to develop a more efficacious therapeutic strategy for such patients, particularly for those with lymph node metastasis and/or concomitant CIS. PMID- 18380819 TI - Complication rates of ultrasound-guided prostate biopsy: a nation-wide survey in Japan. AB - OBJECTIVE: To assess the morbidity secondary to ultrasound-guided systematic prostate biopsy in Japan. METHODS: Five hundred and forty-eight principal urological training institutions certified by the Japanese Urological Association participated in the survey. A retrospective analysis of complications after prostate biopsy was conducted. RESULTS: Medical records of 212 065 procedures carried out between 2004 and 2006 were reviewed. Seventy-six percent of procedures had been carried out transrectally, and 23% transperineally. Hematuria, rectal bleeding and hematospermia occurred in 12%, 5.9% and 1.2% of cases, respectively. Voiding symptoms were reported after biopsy in 1.9% of cases and urinary retention in 1.1%. Fever (>38 degrees C) was observed in 1.1% and sepsis occurred in 0.07%. Hospitalization was required in 0.69% of cases for the treatment of biopsy-related complications. CONCLUSIONS: Overall, contemporary ultrasound-guided systematic prostate biopsy is safely carried out in Japan. However, minor complications are not infrequent, whereas major ones remain very rare but life-threatening if they occur. Thus, adequate information should be provided to patients undergoing prostate biopsy. PMID- 18380820 TI - The potential role of prebiopsy magnetic resonance imaging combined with prostate specific antigen density in the detection of prostate cancer. AB - AIM: Two-thirds of patients with a gray-zone prostate-specific antigen (PSA) level undergo unnecessary biopsy. Sensitivity is not yet sufficient to permit the use of modified PSA parameters or magnetic resonance (MR) imaging alone for prostate cancer screening. Thus, we evaluated the combination of MR imaging and PSA density (PSAD) for specificity and sensitivity. METHODS: During the period April 2004 through March 2006, 185 patients with a PSA level of 4.0-10.0 ng/mL underwent MR imaging and transrectal ultrasonography-guided 8-core biopsy (systemic sextant biopsy of the peripheral zone plus two cores of transition zone). All MR images were interpreted prospectively by two radiologists. An image was considered positive for prostate cancer if any feature indicated a cancerous lesion. Receiver operating characteristic (ROC) curves were used to compare the usefulness of the PSA level, PSAD and PSA transitional zone density (PSATZ) for the detection of prostate cancer. RESULTS: Of the 185 patients, 62 had prostate cancer. Sensitivity and specificity of the axial T2-weighted MR imaging findings for cancer detection were 79.0% and 59.4%, respectively. The area under the ROC curve was 0.590 for the PSA level, 0.718 for PSAD and 0.695 for PSATZ. MR imaging findings and PSAD were shown by multivariate analysis to be statistically significant independent predictors of prostate cancer (P < 0.001). With a PSAD cut-off value of 0.111, sensitivity was 96.8%, but specificity was 19.5%. Combining MR imaging findings with PSAD increased the specificity to 40% and retained 95% sensitivity. CONCLUSION: MR imaging findings combined with PSAD provide high sensitivity and improve the specificity for the early detection of prostate cancer. PMID- 18380822 TI - Prospective comparative study of single dose versus 3-day administration of antimicrobial prophylaxis in minimum incision endoscopic radical prostatectomy. AB - OBJECTIVE: From the critical stand point against the overuse of antimicrobial agents, appropriate reduction of antimicrobial prophylaxis (AMP) should be considered. We have prospectively reduced AMP and evaluated the occurrence of surgical site infection (SSI) following radical retropubic prostatectomy (RRP) by minimum incision endoscopic surgery (MIES). METHODS: A total of 101 consecutive patients who underwent MIES-RRP for prostate carcinoma were classified into two groups according to AMP dose. The 3-day group of 52 patients received tazobactam sodium/piperacillin sodium (TAZ/PIPC) 2.5 g intravenously before the operation and continued twice daily until postoperative day 2, and the single dose group of 49 patients received TAZ/PIPC 2.5 g intravenously only once before the operation. Additional antimicrobial agents were given only when SSI occurred. The occurrence of SSI and remote infection (RI) were analyzed. RESULTS: There was no significant difference in the rate of SSI occurrence between the 3-day group (3.8%) and single dose group (6.1%) (P = 0.6). RI did not increase in the single dose group. CONCLUSION: Antimicrobial prophylaxis dose was successfully reduced without increasing SSI or RI. A single dose of AMP is feasible to prevent SSI and RI and would be a standard regimen in MIES-RRP. Active surveillance of postoperative infection is mandatory to promptly administer antimicrobial treatment as the need arises. PMID- 18380823 TI - Benign prostatic hyperplasia: clinical benefits on Three-Dimensional Ultrasound eXtended Imaging (3D-XI). AB - OBJECTIVES: To prospectively evaluate whether trans-rectal Three-Dimensional eXtended Imaging (3D XI) allows characterization of the prostate gland in cases of isolated benign prostate hyperplasia (BPH) so as to elucidate the motive for discrepancy of postvoiding residual urine. METHODS: The study was conducted according to the standards of the local ethics committee. Patients gave informed consent. Disclosing the 3D XI display of the prostatic urethra and prostatic gland zones was a preliminary essential. The study included 113 men with a clinical diagnosis of BPH in whom transurethral resection of the prostate was planned. Patients aged from 52 to 75 years (mean 63). Other causes of infravesical obstructive uropathy and prostate neoplastic involvement were excluded. All patients were evaluated by three-dimensional Trans-Rectal Ultrasound using 3D XI methods. Patients were grouped according to the postvoiding residual urine volume into three groups, less and more than 100 mL and urine retention. RESULTS: 3D XI provided excellent resolution and diagnostic authority of prostate gland anatomy and for the appraisal of BPH morphology. The balance and type of nodular eruption proved responsible for the severity of symptoms aspires by extra-axial nodular effect upon the prostate urethra. The 3D XI analysis regarding the nodular stromal to glandular ratio compared to the histopathological results proved effectual in 98.3% of stromal-dominant, 100% in glandular-dominant and 93.3% in mixed type hyperplasia. CONCLUSIONS: The severity of symptoms among men with BPH showed an association with the nodular credence, highlighted by 3D XI as a supportive tool in characterizing BPH. PMID- 18380824 TI - Prevalence of chronic prostatitis symptoms in a randomly surveyed adult population of urban-community-dwelling Nigerian males. AB - OBJECTIVES: Chronic prostatitis is a debilitating disease of the prostate gland that is characterized by chronic pelvic pain. Unlike the developed world, the prevalence of chronic prostatitis and its impact on the quality of life (QOL) of the sufferer is not known in Nigeria. This work was carried out to fill this vacuum. METHODS: A random cross-sectional survey of apparently normal men of ages 20-70 years, living in Nsukka, a university town in South-eastern Nigeria, using the National Institute of Health Chronic Prostatitis Symptom Index (NIH-CPSI) was carried out. The self reported pain and QOL scores were used to identify subjects with chronic prostatitis symptoms and the subjects' QOL status, respectively. RESULTS: Of the 1507 men studied, 12.21% had symptoms suggestive of chronic prostatitis. Subjects with chronic prostatitis symptoms (CPS) (mean relative age 40.98 +/- 12.51 years) had higher mean pain and voiding scores (5.65 +/- 1.99 and 2.61 +/- 2.23, respectively) compared with subjects without CPS (0.44 +/- 0.73 and 1.29 +/- 1.70 for pain and voiding scores, respectively; mean relative age 41.99 +/- 12.99 years). A total of 39.62% of the population had pain in at least one location, while 22.69% of the population had impaired QOL. CONCLUSION: Chronic prostatitis symptoms are as common in Nigeria as they are in the developed world, and so are their effects on the QOL of the sufferer. PMID- 18380825 TI - Impact of intravenous furosemide on flow rate characteristics and clinic waiting times. AB - OBJECTIVE: To assess the impact of a single dose of i.v. furosemide on clinic waiting times and urinary flow rate parameters in a group of symptomatic men. METHODS: Sixty-two consecutive men attending a prostate assessment clinic were prospectively entered into the study. They were then randomized to receive 10 mg of i.v. furosemide or to drink oral fluids only, as per standard protocol. Flow rate parameters and clinic waiting times were recorded. RESULTS: The clinic waiting time was significantly reduced by the administration of i.v. furosemide compared to oral fluid alone (81 vs 36 min; P < 0.0001) without any alteration in flow rate characteristics. Interestingly, although there was no statistically significant difference between the voided volumes in both groups (251 vs 313 mL; P = 0.115) all of the patients given furosemide voided a volume of over 150 mL, where as almost a third (10/32) of those receiving oral fluid alone did not achieve this. There were no adverse events following the administration of i.v. diuretic. CONCLUSION: A single dose of i.v. furosemide dramatically reduces the overall clinic waiting time without inducing statistically significant changes in flow rate parameters. Indeed, none of the patients receiving furosemide voided less than 150 mL. We would thus recommend this practice be considered as an alternative to standard uroflowmetry protocols. PMID- 18380826 TI - Duration of erectile dysfunction and its relationship to treatment seeking and satisfaction with treatment using PDE5 inhibitors. AB - OBJECTIVES: This cross-sectional study was designed to evaluate whether the duration of erectile dysfunction (ED) influenced treatment seeking and satisfaction with treatment using PDE5 inhibitors. METHODS: Participants were 409 men with ED who were primarily recruited over the internet via men's health web sites. Participants completed a questionnaire to assess the duration and perceived severity of ED, information and help-seeking behaviors for ED, and treatment usage and satisfaction with PDE5 inhibitor medication. RESULTS: The results demonstrated that men with ED of longer duration were more likely to have discussed their ED with their partner and doctor and to have sought information and treatment for their ED problem. No differences were found in reported satisfaction with ED medication usage or expected future medication use across the varying levels of ED duration, once variance attributable to age was accounted for. CONCLUSIONS: These results suggest that men are more likely to accept that they have ED and seek treatment for their ED with increasing duration of the condition, although these men are not more satisfied with PDE5 inhibitors as a treatment option. PMID- 18380827 TI - Verapamil prevents the apoptotic and hemodynamic changes in response to unilateral ureteral obstruction. AB - INTRODUCTION: Obstruction of the urinary tract has marked effects on renal blood flow, glomerular filtration rate (GFR), and tubular function. Moreover, ureteral obstruction results in an injury response that can progress to irreversible renal fibrosis and tubular atrophy by apoptosis. METHODS: We examined the effect of a calcium channel blocker (verapamil) on renal functions and the abundance of apoptotic (p53, Fas, proliferating cell nuclear antigen [PCNA]) markers 1 week after Unilateral Ureteral Obstruction (UUO). RESULTS: Immunohistochemistry studies revealed that UUO was markedly associated with up-regulation in the expression of p53 (1550 +/- 82 vs 100 +/- 23%), Fas (657 +/- 48 vs 100 +/- 31%), and proliferating cell nuclear antigen (945 +/- 70 vs 100 +/- 17% of sham levels). Administration of verapamil normalized the up-regulation of apoptotic markers p53 (724 +/- 116 vs 1550 +/- 82%); Fas (162 +/- 38 vs 657 +/- 48%) and PCNA (353 +/- 54 vs 945 +/- 70%). Furthermore, tubular diameter, as an important marker for detecting tubular atrophy was significantly decreased compared to those in UUO rabbits. The percent area of interstitial fibrosis in UUO kidneys was significantly greater than that in Verapamil-treated kidneys. Importantly, Verapamil reduced the development of interstitial fibrosis in UUO rabbits. We measured the GFR and renal blood flow in UUO. Short-term Verapamil challenge partially prevented the decrease in GFR (non-treated UUO: 62 +/- 14; Verapamil + UUO: 119 +/- 7; Sham: 127 +/- 23 microL x min(-1) x kg body wt(-1), P < 0.05) and renal blood flow (non-treated UUO: 1.1 +/- 0.4; Verapamil + UUO: 5.0 +/- 0.2; sham: 6.3 +/- 0.2 mL x min(-1) x kg body wt(-1), P < 0.05). CONCLUSION: Verapamil significantly prevents impairment in renal function and also prevents the up regulation of p53, Fas, and PCNA during UUO, demonstrating a marked renoprotective effect of Verapamil treatment in conditions with urinary tract obstruction. PMID- 18380828 TI - Effect of eviprostat on bladder overactivity in an experimental cystitis rat model. AB - OBJECTIVE: The purpose of this study was to investigate the effects of eviprostat, a phytotherapeutic drug, on bladder overactivity and inflammation in a cyclophosphamide (CYP)-induced cystitis rat model. METHODS: Female Sprague Dawley rats received a single intraperitoneal injection of CYP (200 mg/kg) or saline. After the CYP injection, eviprostat (9, 18 or 54 mg/kg per day) or a vehicle was orally given twice each day. Four days after the CYP injection, bladder function was evaluated by cystometrograms under urethane anesthesia. In a separate group, bladder inflammation was compared between the eviprostat- or vehicle-treated animals. Furthermore, the effects of eviprostat on carbachol induced muscle contraction were evaluated by an in vitro experiment. RESULTS: The intercontraction interval (ICI) significantly decreased in the CYP-injected rats in comparison to the saline-injected rats. In the CYP-injected group, 18 and 54 mg/kg per day of eviprostat treatment significantly increased the ICI, but did not change the maximum voiding pressure in comparison to the vehicle treatment. In the saline-injected group, no significant changes of any parameters in the cystometrograms were observed between the eviprostat- and vehicle-treated groups. CYP-induced bladder inflammation tended to be lower in the eviprostat-treated group in comparison to the vehicle-treated group. An in vitro experiment revealed that eviprostat failed to inhibit carbachol-induced muscle contraction. CONCLUSION: The oral administration of eviprostat suppressed CYP-induced bladder overactivity. The effects of eviprostat on the micturition reflex may be irrespective of antimuscarinic action. The present findings raise the possibility that eviprostat could be an effective treatment for bladder overactivity associated with inflammation. PMID- 18380829 TI - Preoperative management of Cushing's syndrome with metyrapone for severe psychiatric disturbances. AB - A 56-year-old woman suffered from severe depression due to Cushing's syndrome with right adrenal adenoma. She had a normal mental state before developing Cushing's syndrome. Because of her depressive state, informed consent for the treatment could not be obtained, and oral administration of 1.5 g/day metyrapone was commenced. After two weeks, her mental state had markedly improved. She subsequently underwent a right adrenalectomy, and metyrapone administration was terminated on the day of surgery. Without modification on the replacement of corticosteroids, the postoperative course was uneventful. Preoperative management of Cushing's syndrome with metyrapone may be useful in cases of severe psychiatric disturbances due to hypercortisolism. PMID- 18380830 TI - Unusual solitary metastasis of the ciliary body in renal cell carcinoma. AB - Renal cell carcinoma (RCC) usually metastasizes to the lung, liver, bone; ocular metastasis is uncommon. We describe a rare case of metachronous ciliary RCC metastasis in a 42-year-old man who had undergone left radical nephrectomy for conventional RCC (pT3aN0M0, G2 Fuhrman) 6 years earlier. Solitary metastasis of the left eye presented with inflammatory symptoms, but examination of the fundus and bulbar ultrasound revealed a small mass of the ciliary body. Initial radiotherapy was unsuccessful and definitive treatment consisted of ocular enucleation with radical result and no further evidence of local and distant disease. Ocular metastasis of RCC is rare, can appear years after treating the primary tumor and should not be excluded in RCC follow-up. As for other RCC solitary metastasis, the best option remains the radical surgical approach. PMID- 18380831 TI - Metastatic renal cell carcinoma to right ventricle without vena caval involvement. AB - Cardiac metastases from renal cell carcinoma without vena caval involvement are extremely rare. We report 49-year-old man who presented symptoms of heart failure and thrombocytopenia. Computed tomography and echocardiography revealed a left renal tumor and a right ventricular mass without vena caval involvement. His symptoms progressed rapidly and he died at nine days following diagnosis of the right ventricular tumor. PMID- 18380832 TI - BK virus-associated nephropathy in a kidney transplant recipient successfully treated with cidofovir, the first case in Japan. AB - A 51-year-old female received a kidney transplant, donated by her husband. The patient was induced with tacrolimus, mycophenolate mofetil and prednisolone. After methyl prednisolone pulse therapy without biopsy, allograft biopsy on POD 160 showed severe tubulo-interstitial nephritis with intranuclear inclusions. Urine cytology also showed decoy cells. Blood PCR detected an increase of BK virus DNA. She was diagnosed as having BK virus-associated nephropathy . Reduction of tacrolimus and switching of mycophenolate mofetil to mizoribine were done. Serum Creatinin (sCr) still rose to 3.0 mg/dl with persistent viremia and viruria. From on POD 268, 0.25 mg/kg of cidofovir was administered intravenously every two weeks over about four months. Biopsy on POD 387 revealed the disappearance of tubulitis with intranuclear inclusions, and decoy cells also disappeared from urine cytology. BK virus DNA in the blood decreased under the threshold level. sCr was stable and remained about 2.2 mg/dl for three months after the final treatment of cidofovir. PMID- 18380833 TI - Fournier gangrene associated with hyper IgE syndrome (Job syndrome). AB - We report a case of a 32-year-old man with hyper IgE syndrome (Job syndrome) who developed Fournier gangrene due to infectious multiple atheromas of the scrotal skin that progressed to the right groin and thigh. The patient required surgical debridement and subsequent skin grafting. This is a rare case of Fournier gangrene associated with hyper IgE syndrome (Job syndrome). When a patient without diabetes mellitus has repeated infections and atopic-like dermatitis, Job syndrome should be considered. PMID- 18380834 TI - Paratesticular angioleiomyoma with cytological atypia. AB - Angioleiomyomas are common benign smooth muscle neoplasms in the dermis and subcutaneous tissue of limbs. Very rarely have they been described in the external male genitalia with only two cases in the paratesticular tissues and another intratesticular. We report a case of a paratesticular angioleiomyoma with cytological atypia and a peculiar vascular pattern, found incidentally as a painless nodule in a 75-year-old man. The main clinicopathological features together with the differential diagnoses and the pertinent published reports are reviewed. PMID- 18380836 TI - Resin bonding to ceramic. PMID- 18380837 TI - Indirect implant-supported fixed provisional restoration in the esthetic zone: fabrication technique and treatment workflow. AB - Treatment objectives of an implant-supported fixed provisional restoration include shaping/preservation of the gingival soft tissue contour, functional and esthetic substitution of the missing dentition during postsurgical healing, and definitive prosthesis fabrication stages. Fixed provisional restoration can also serve as an esthetic and functional blueprint in the fabrication of the definitive restoration. Despite its common use and important indications, limited information is available on the various aspects of the provisional fabrication and treatment. This article presents a production technique and treatment workflow of a laboratory-fabricated, screw-retained fixed provisional restoration. Provisional restoration is fabricated using layering technique and internal stain characterization. The soft tissue profile of the working cast is modified according to the coronal contour of the diagnostic wax-up. Upon delivery, the provisional contour is reevaluated and modified as necessary. The developed emergence profile of the provisional restoration is transferred to the master cast via customized impression copings. CLINICAL SIGNIFICANCE: Laboratory fabricated implant-supported provisional restorations allow the esthetic and functional substitution of the missing dentition and the shaping of the soft tissue profile, and can act as a blueprint in the fabrication of definitive restorations. PMID- 18380839 TI - Rotational path removable partial denture (RPD): conservative esthetic treatment option for the edentulous mandibular anterior region: a case report. AB - It can be esthetically and financially daunting for patients to lose teeth in an anterior region of the mouth. For these patients, traditional treatment options presented in the past have included fixed partial denture, implants, and conventional removable partial denture (RPD). For patients faced with financial, anatomical, and/or esthetic limitations, the edentulous region can be restored successfully with a rotational path RPD. Rotational path RPD designs have often been overlooked by the dental profession due to its complex concepts involving the prosthetic design and sensitive laboratory techniques. With better understanding of the concepts and design, the dental clinician can deliver the highest esthetic outcome in compromised areas in which other treatment options may often face limitations. This paper reviews the method used to esthetically design and plan a posterior-anterior rotational path RPD in an edentulous mandibular anterior region for a patient missing the mandibular incisors. CLINICAL SIGNIFICANCE: Due to inadequate understanding of the mechanics of rotational path RPDs, many clinicians have not adapted the application of this advantageous prosthesis. When correctly designed and fabricated, the rotational path RPD provides improved esthetics, cleanliness, and retention for patients who may not be suitable candidates for implants or fixed partial dentures in tooth supported edentulous regions. PMID- 18380841 TI - Power distribution across the face of different light guides and its effect on composite surface microhardness. AB - PURPOSE: The aim of this study was to examine the influence of light guide type on the distribution of irradiant emission from a light-emitting diode (LED) curing light and to measure the effect of light dispersion on surface microhardness across the top and bottom surfaces of two types of composite resins. MATERIALS AND METHODS: A laser beam analyzer (LBA-700, Spiricon) was used to evaluate light distribution (Top Hat factor [THF]) across the distal surface of a standard and turbo light guide from an LED curing light (Bluephase, Ivoclar Vivadent). Composites (Z100 [hybrid], A110 [microfill]; 3M ESPE) were placed in blackened rings (2 x 11 mm) and exposed at 0 mm for 5 seconds (Z100) or 15 seconds (A110) using the light guides at similar irradiance, energy density, and exit diameters (N = 5). Similar irradiance values were produced by using the turbo light guide on the "low power" setting of the curing light and the "high power" setting when using the standard light guide. THF values were analyzed with an unpaired t-test. Knoop hardness (KHN) was determined on the top and bottom surfaces (Leco) in 1-mm lateral increments from the specimen center and proceeding 4 mm in both east-west and north-south directions. The effects of the major factors (light guide type and lateral distance) on the hardness of each composite were analyzed using multiple analysis of variance (ANOVA), and a two tailed, unpaired Dunnett's t-test determined when lateral hardness values significantly differed from that at the specimen center. The percentage difference between maximum and minimum (max-min) hardness values for each specimen, with respect to distance from specimen center, and the percentage decrease for the standard and turbo light guides, with respect to both composite resin types, were compared using ANOVA and the Tukey's post-hoc test. RESULTS: The standard guide had a significantly higher (i.e., more uniform light distribution) THF than did the turbo tip (p < 0.001). For the microfill, significant differences in hardness were found based on the distance from the specimen center (p < 0.0001), and with respect to the top or bottom surfaces (p < 0.0001). However, no difference was found between the two types of light guides (p = 0.939). For the hybrid, significant differences in hardness were found based on lateral distance (p < 0.001), surface (p < 0.001), and light guide type (p = 0.045).However, for both composites, significant interactions were present. Significant differences were found between hardness at the specimen center and at various lateral distances, depending on composite type, surface, and light guide type. The percent age max-min hardness decrease across the surface was significantly less for the standard light guide (p < 0.0001) and at the top surface (p < 0.02) with both composite resin types. Also, the least percentage max-min hardness decrease occurred on the top surface of the microfill material (p < 0.001). CONCLUSION: The standard light guide produced a more homogeneous distribution of light across the tip end compared with the turbo light guide, based on the THF. Composite surface hardness patterns correlated with the applied irradiance distribution profiles, yielding greater hardness at higher irradiance locations for both top and bottom surfaces. CLINICAL SIGNIFICANCE: Light guide selection may influence the uniformity of surface and subsurface hardness when light-curing composite resin restorations. PMID- 18380843 TI - Influence of differently oriented dentin surfaces and the regional variation of specimens on adhesive layer thickness and bond strength. AB - STATEMENT OF THE PROBLEM: Adhesive systems can spread differently onto a substrate and, consequently, influence bonding. PURPOSE: The purpose of this study was to evaluate the effect of differently oriented dentin surfaces and the regional variation of specimens on adhesive layer thickness and microtensile bond strength (MTBS). MATERIALS AND METHODS: Twenty-four molars were sectioned mesiodistally to expose flat buccal and lingual halves. Standardized drop volumes of adhesive systems (Single Bond [SB] and Prime & Bond 2.1 [PB2.1]) were applied to dentin according to the manufacturer's instructions. Teeth halves were randomly divided into groups: 1A-SB/parallel to gravity; 1B-SB/perpendicular to gravity; 2A-PB2.1/parallel to gravity; and 2B-PB2.1/perpendicular to gravity. The bonded assemblies were stored in 37 degrees C distilled water for 24 hours and then sectioned to obtain dentin sticks (0.8 mm2). The adhesive layer thickness was determined in a light microscope (x200), and after 48 hours the specimens were subjected to MTBS test. Data were analyzed by one-way and two-way analysis of variance and Student-Newman-Keuls tests. RESULTS: Mean values (MPa +/- SD) of MTBS were: 39.1 +/- 12.9 (1A); 32.9 +/- 12.4 (1B); 52.9 +/- 15.2 (2A); and 52.3 +/- 16.5 (2B). The adhesive systems' thicknesses (microm +/- SD) were: 11.2 +/- 2.9 (1A); 18.1 +/- 7.3 (1B); 4.2 +/- 1.8 (2A); and 3.9 +/- 1.3 (2B). No correlation between bond strength and adhesive layer thickness for both SB and PB2.1 (r = -0.224, p = 0.112 and r = 0.099, p = 0.491, respectively) was observed. CONCLUSIONS: The differently oriented dentin surfaces and the regional variation of specimens on the adhesive layer thickness are material-dependent. These variables do not influence the adhesive systems' bond strength to dentin. CLINICAL SIGNIFICANCE Adhesive systems have different viscosities and spread differently onto a substrate, influencing the bond strength and also the adhesive layer thickness. Adhesive thickness does not influence dentin bond strength, but it may impair adequate solvent evaporation, polymer conversion, and may also determine water sorption and adhesive degradation over time. In the literature, many studies have shown that the adhesive layer is a permeable membrane and can fail over time because of its continuous plasticizing and degradation when in contact with water. Therefore, avoiding thick adhesive layers may minimize these problems and provide long-term success for adhesive restorations. PMID- 18380845 TI - Direct dentin bonding technique sensitivity when using air/suction drying steps. AB - STATEMENT OF PROBLEM: Moisture control before and after application of the primer/adhesive components of etch-and-rinse dentin bonding agents is usually achieved using a stream of air delivered by an air syringe. Suction drying with a suction tip is a common alternative for moisture control, but data about the use of suction drying instead of the air syringe is scarce or nonexistent. PURPOSE: The purpose of this study was to compare the dentin microtensile bond strength (MTBS) using either the air syringe or the suction tip to control the amount of moisture. MATERIALS AND METHODS: Fifteen freshly extracted human molars were divided randomly into three groups of five. A three-step etch-and-rinse dentin bonding agent (OptiBond FL) was used. Group 1 was the control group and utilized air drying alone (with an air syringe) during the placement of the dentin adhesive on the ground-flat occlusal dentin surface. Group 2 also used air drying alone, but teeth were prepared with a standardized MOD cavity. Group 3 utilized suction drying alone in the standardized MOD cavity. All teeth were restored with 1.5-mm-thick horizontal increments of composite resin (Filtek Z100). Specimens were stored in water for 24 hours, then prepared for a nontrimming MTBS test. Bond strength data were analyzed with a Kruskal-Wallis test at p < 0.05. Specimens were also evaluated for mode of fracture and interface characterization using scanning electron microscope (SEM) analysis. RESULTS: The mean MTBSs were not statistically different from one another (p = 0.54) at 54.0 MPa (air-drying, flat dentin), 53.4 MPa (air-drying, MOD), and 49.2 MPa (suction drying, MOD). Microscopic evaluation of failure modes indicated that most failures were interfacial. Failed interfaces, when analyzed under SEM, appeared typically mixed with areas of failed adhesive resin and areas of cohesively failed dentin. CONCLUSIONS: There are no differences in MTBS to human dentin using either the air syringe or the suction tip to control the amount of moisture. The conventional three-step dentin bonding agent used in the present study not only proved insensitive to the moisture-control method but also to the effect of increased polymerization shrinkage stress (ground-flat versus MOD preparation). CLINICAL SIGNIFICANCE: Although the effect of common errors on the performance of total-etch adhesives has been investigated, data about the use of suction drying instead of an air syringe is scarce or nonexistent. The present study demonstrated that both the air syringe and the suction tip can be used to control moisture when using etch-and-rinse dentin bonding agents. The conventional three step dentin bonding agent tested, OptiBond FL, demonstrated low technique sensitivity. PMID- 18380847 TI - Critical appraisal: effects of bleaching on tooth structure and restorations, part III: effects on dentin. PMID- 18380849 TI - Abstracts of the 2008 Annual Scientific Meeting of the American Geriatrics Society. May 1-3, 2008. PMID- 18380848 TI - Meth mouth: methamphetamine and oral health. PMID- 18380850 TI - Impact of a commercial destocking relief intervention in Moyale district, southern Ethiopia. AB - A commercial destocking intervention was piloted in southern Ethiopia during the drought of early 2006. The intervention led to the purchase of an estimated 20,000 cattle valued at USD 1.01 million. On average, destocked households received USD 186 from the sale of cattle--approximately 5,405 households were involved. In terms of aid investment, the approximate cost-benefit ratio was 41:1. During the drought, income from destocking accounted for 54.2 per cent of household income (n = 114 households), and was used to buy food, care for livestock, meet various domestic expenses, support relatives, and either pay off debts or augment savings. Seventy-nine per cent of the income derived from destocking was used to buy local goods or services. Expenditure on livestock care amounted to 36.5 per cent of local spending, and included the private transportation of livestock to better grazing areas. The buoyant livestock export trade was considered to be an important driver of commercial destocking, demonstrating a positive link between livestock and meat exports, and pastoral vulnerability during drought. PMID- 18380851 TI - Changing household responses to drought in Tharaka, Kenya: vulnerability, persistence and challenge. AB - Drought is a recurring challenge to the livelihoods of those living in Tharaka District, Kenya, situated in the semi-arid zone to the east of Mount Kenya, from the lowest slopes of the mountain to the banks of the Tana River. This part of Kenya has been marginal to the economic and political life of Kenya from the colonial period until the present day. A study of more than 30 years of change in how people in Tharaka cope with drought reveals resilience in the face of major macro-level transformations, which include privatisation of landownership, population growth, political decentralisation, increased conflict over natural resources, different market conditions, and environmental shifts. However, the study also shows troubling signs of increased use of drought responses that are incompatible with long-term agrarian livelihoods. Government policy needs to address the challenge of drought under these new macro conditions if sustainable human development is to be achieved. PMID- 18380852 TI - 'It'll never happen to me': understanding public awareness of local flood risk. AB - Following the severe flood events of 1998 and 2000, the United Kingdom's Environment Agency prioritised the need to increase public flood risk awareness. Drawing on data collected during research undertaken for the Environment Agency, this paper contributes to understanding of one aspect of flood awareness: people's recognition that their property is in an area that is potentially at risk of flooding. Quantitative analyses indicate that class is the most influential factor in predicting flood risk awareness, followed by flood experience and length of time in residence. There are also significant area differences. Our qualitative work explores how those defined as 'at risk' account for their lack of awareness or concern about their risk status. We conclude that the problem is often not simply a lack of awareness, but rather, assessments of local risk based on experience that underestimate the impact of rare or extreme events. We underline the importance of engaging with local perspectives on risk and making local people part of 'awareness-raising' processes. PMID- 18380853 TI - Collaborative emergency management: better community organising, better public preparedness and response. AB - Community coordination requires communication and planning of precautions to take when faced with a severe threat of disaster. The unique case of the four Florida hurricanes of 2004--Charley, Frances, Ivan, and Jeanne--is used here to assess community responses to repeated threats of hurricanes. The paper examines how effectiveness in coordinating community disaster response efforts affects future public preparedness. The findings suggest that pre-season planning, open communication between emergency managers and elected officials, and the use of technology all had a significant impact on community responses. The repeated threat scenario indicates that emergency managers must work vigilantly to keep residents informed of the seriousness of a situation. The study describes how emergency managers in Florida countered public complacency during four hurricanes in six weeks. The strategies identified as useful by public managers in the context of hurricanes are applicable to other natural and man-made disasters. PMID- 18380854 TI - Understanding the security management practices of humanitarian organizations. AB - Humanitarian organisations operate in increasingly hostile environments. Although authoritative statistics are scarce, anecdotal evidence suggests that aid workers face life-threatening risks that are exacerbated by the growing number of humanitarian organisations operating in the field, the diversity of their mandates, the lack of common professional security standards, and limited success in inter-agency security coordination. Despite broad acceptance of the need for better security management and coordination, many humanitarian organisations remain ambivalent about devoting increased resources to security management and security coordination. A critical lack of basic empirical knowledge of the field security environment hampers efforts to enhance security management practices. The absence of a systematic means of sharing incident data undermines the capacity of the humanitarian community to address proactively security threats. In discussions about humanitarian staff safety and security, the least common denominator remains cumulative anecdotal evidence provided by the many security personnel working for humanitarian organisations in the feld. PMID- 18380855 TI - Psychological consequences among adults following the 1999 earthquake in Athens, Greece. AB - One year after the 7 September 1999 earthquake in Athens, Greece, we investigated the psychological consequences among 305 individuals (71 per cent female) residing in the settlements of Ano Liosia Municipality. Adaptability was difficult (63 per cent) due to limited space (50 per cent). Insecurity feelings were predictive of difficult adaptability (chi2= 29.8, p<0.0001) and were common (63 per cent) among married subjects, independent of age (chi2= 5.0, odds ratio (OR): 0.49, 95 per cent confidence interval (CI): 0.26-0.90). Eighty per cent expressed stress feelings, mainly nervousness/tension (60 per cent). Adaptability (chi2= 5.3, OR: 0.5, 95 per cent CI: 0.27-0.9), age (chi2= 6.5, OR: 1.03, 95 per cent CI: 1.01-1.06), and female gender (chi2= 4.7, OR: 0.48, 95 per cent CI: 0.25 0.90) were independent predictors of stress feelings. The majority (55 percent) developed sleep disorders, chiefly insomnia (60 percent). Adaptability problems were the only predictor of sleep disorders (chi2= 6.4, OR: 0.5, 95 per cent CI: 0.33-0.87). Psychiatric medication use increased after the earthquake. PMID- 18380856 TI - Decision making as community adaptation: a case study of emergency managers in Oklahoma. AB - This paper explores how emergency managers make judgments regarding long-term policy and offers a sociological account of organisational decision making within an ecological context. Discussions with emergency managers focusing on the relative merits of rainfall estimation and tornado detection served as data with which to address these issues. Among the 39 interviewees, a consensus emerged favouring tornado detection over rainfall estimation. From these findings, the paper attempts to understand why emergency managers prefer tornado detection to rainfall estimation and to develop theoretical generalisations explaining trends in these preferences. When developing long-term policy, analysis of transcripts revealed emergency managers to be most concerned with the relative uncertainty of hazards, the capabilities of technology in hazard mitigation, and how the public perceives environmental threats. Given the environmental, technological, and social concerns reflected in this reasoning, there appears to be a strong ecological context driving the need for tornado detection among emergency managers. Implications and concerns are presented in the final section. PMID- 18380857 TI - Overview of deaths associated with natural events, United States, 1979-2004. AB - Analysis of the National Center for Health Statistics' Compressed Mortality File showed that between 1979 and 2004, natural events caused 21,491 deaths in the United States. During this 26-year period, there were 10,827 cold-related deaths and 5,279 heat-related deaths. Extreme cold or heat accounted for 75 per cent of the total number of deaths attributed to natural events--more than all of deaths resulting from lightning, storms and foods, and earth movements, such as earthquakes and landslides. Cold-related death rates were highest in the states of Alaska, Montana, New Mexico, and South Dakota, while heat-related deaths were highest in the states of Arizona, Missouri, and Arkansas. These deaths occurred more often among the elderly and black men. Other deaths were attributed to lightning (1,906), storms and foods (2,741), and earth movements (738). Most deaths associated with natural events are preventable and society can take action to decrease the morbidity and mortality connected with them. PMID- 18380858 TI - Returning home: resettlement of formerly abducted children in Northern Uganda. AB - This exploratory qualitative study considers the subjective resettlement experiences of children forced into armed conflict in Northern Uganda from the perspectives of 11 former child combatants and 11 adult community members. A thematic analysis was performed on the narrative data. The bioecological model was used to provide a conceptual framework for key themes. Major findings included the overarching impact of ongoing armed conflict on returnees' lives, the important role of the family in supporting children's resettlement, the harassment of former child soldiers by community members, and the community's inability to support systematically the returning children in tangible ways. This study recommends that humanitarian services at all levels strengthen the capacity of families to care for the material and psychoemotional needs of former child soldiers within their communities. PMID- 18380859 TI - Symposium on 'human evolution: ancestors and relatives'. PMID- 18380861 TI - The hominin fossil record: taxa, grades and clades. AB - This paper begins by reviewing the fossil evidence for human evolution. It presents summaries of each of the taxa recognized in a relatively speciose hominin taxonomy. These taxa are grouped in grades, namely possible and probable hominins, archaic hominins, megadont archaic hominins, transitional hominins, pre modern Homo and anatomically modern Homo. The second part of this contribution considers some of the controversies that surround hominin taxonomy and systematics. The first is the vexed question of how you tell an early hominin from an early panin, or from taxa belonging to an extinct clade closely related to the Pan-Homo clade. Secondly, we consider how many species should be recognized within the hominin fossil record, and review the philosophies and methods used to identify taxa within the hominin fossil record. Thirdly, we examine how relationships within the hominin clade are investigated, including descriptions of the methods used to break down an integrated structure into tractable analytical units, and then how cladograms are generated and compared. We then review the internal structure of the hominin clade, including the problem of how many subclades should be recognized within the hominin clade, and we examine the reliability of hominin cladistic hypotheses. The last part of the paper reviews the concepts of a genus, including the criteria that should be used for recognizing genera within the hominin clade. PMID- 18380860 TI - Reconstructing phylogenies and phenotypes: a molecular view of human evolution. AB - This review broadly summarizes how molecular biology has contributed to our understanding of human evolution. Molecular anthropology began in the 1960s with immunological comparisons indicating that African apes and humans were closely related and, indeed, shared a common ancestor as recently as 5 million years ago. Although initially dismissed, this finding has proven robust and numerous lines of molecular evidence now firmly place the human-ape divergence at 4-8 Ma. Resolving the trichotomy among humans, chimpanzees and gorillas took a few more decades. Despite the readily apparent physical similarities shared by African apes to the exclusion of modern humans (body hair, knuckle-walking, thin tooth enamel), the molecular support for a human-chimpanzee clade is now overwhelming. More recently, whole genome sequencing and gene mapping have shifted the focus of molecular anthropology from phylogenetic analyses to phenotypic reconstruction and functional genomics. We are starting to identify the genetic basis of the morphological, physiological and behavioural traits that distinguish modern humans from apes and apes from other primates. Most notably, recent comparative genomic analyses strongly indicate that the marked differences between modern humans and chimpanzees are likely due more to changes in gene regulation than to modifications of the genes themselves, an idea first proposed over 30 years ago. Almost weekly, press releases describe newly identified genes and regulatory elements that seem to have undergone strong positive selection along the human lineage. Loci involved in speech (e.g. FOXP2), brain development (e.g. ASPM), and skull musculature (e.g. MYH16) have been of particular interest, but some surprising candidate loci (e.g. those involved in auditory capabilities) have emerged as well. Exciting new research avenues, such as the Neanderthal Genome Project, promise that molecular analyses will continue to provide novel insights about our evolution. Ultimately, however, these molecular findings can only be understood in light of data from field sites, morphology labs, and museum collections. Indeed, molecular anthropology depends on these sources for calibrating molecular clocks and placing genetic data within the context of key morphological and ecological transitions in human evolution. PMID- 18380862 TI - The environmental context of human evolutionary history in Eurasia and Africa. AB - This review has three main aims: (1) to make specific predictions about the habitat of the hypothetical last common ancestor of the chimpanzee/bonobo-human clade; (2) to outline the major trends in environments between 8-6 Ma and the late Pleistocene; and (3) to pinpoint when, and in some cases where, human ancestors evolved to cope with the wide range of habitats they presently tolerate. Several lines of evidence indicate that arboreal environments, particularly woodlands, were important habitats for late Miocene hominids and hominins, and therefore possibly for the last common ancestor of the chimpanzee/bonobo-human clade. However, as there is no clear candidate for this last common ancestor, and because the sampling of fossils and past environments is inevitably patchy, this prediction remains a working hypothesis at best. Nonetheless, as a primate, it is expected that the last common ancestor was ecologically dependent on trees in some form. Understanding past environments is important, as palaeoenvironmental reconstructions provide the context for human morphological and behavioural evolution. Indeed, the impact of climate on the evolutionary history of our species has long been debated. Since the mid-Miocene, the Earth has been experiencing a general cooling trend accompanied by aridification, which intensified during the later Pliocene and Pleistocene. Numerous climatic fluctuations, as well as local, regional and continental geography that influenced weather patterns and vegetation, created hominin environments that were dynamic in space and time. Behavioural flexibility and cultural complexity were crucial aspects of hominin expansion into diverse environments during the Pleistocene, but the ability to exploit varied and varying habitats was established much earlier in human evolutionary history. The development of increasingly complex tool technology facilitated re-expansion into tropical forests. These environments are difficult for obligate bipeds to negotiate, but their exploitation was accomplished by archaic and/or anatomically modern humans independently in Africa and south-east Asia. Complex social behaviour and material culture also allowed modern humans to reach some of the most hostile regions of the globe, above the Arctic Circle, by the late Pleistocene. This, with colonization of the Americas and Australasia, established Homo sapiens as a truly cosmopolitan species. PMID- 18380863 TI - Hominin life history: reconstruction and evolution. AB - In this review we attempt to reconstruct the evolutionary history of hominin life history from extant and fossil evidence. We utilize demographic life history theory and distinguish life history variables, traits such as weaning, age at sexual maturity, and life span, from life history-related variables such as body mass, brain growth, and dental development. The latter are either linked with, or can be used to make inferences about, life history, thus providing an opportunity for estimating life history parameters in fossil taxa. We compare the life history variables of modern great apes and identify traits that are likely to be shared by the last common ancestor of Pan-Homo and those likely to be derived in hominins. All great apes exhibit slow life histories and we infer this to be true of the last common ancestor of Pan-Homo and the stem hominin. Modern human life histories are even slower, exhibiting distinctively long post-menopausal life spans and later ages at maturity, pointing to a reduction in adult mortality since the Pan-Homo split. We suggest that lower adult mortality, distinctively short interbirth intervals, and early weaning characteristic of modern humans are derived features resulting from cooperative breeding. We evaluate the fidelity of three life history-related variables, body mass, brain growth and dental development, with the life history parameters of living great apes. We found that body mass is the best predictor of great ape life history events. Brain growth trajectories and dental development and eruption are weakly related proxies and inferences from them should be made with caution. We evaluate the evidence of life history-related variables available for extinct species and find that prior to the transitional hominins there is no evidence of any hominin taxon possessing a body size, brain size or aspects of dental development much different from what we assume to be the primitive life history pattern for the Pan-Homo clade. Data for life history-related variables among the transitional hominin grade are consistent and none agrees with a modern human pattern. Aside from mean body mass, adult brain size, crown and root formation times, and the timing and sequence of dental eruption of Homo erectus are inconsistent with that of modern humans. Homo antecessor fossil material suggests a brain size similar to that of Homo erectus s. s., and crown formation times that are not yet modern, though there is some evidence of modern human-like timing of tooth formation and eruption. The body sizes, brain sizes, and dental development of Homo heidelbergensis and Homo neanderthalensis are consistent with a modern human life history but samples are too small to be certain that they have life histories within the modern human range. As more life history-related variable information for hominin species accumulates we are discovering that they can also have distinctive life histories that do not conform to any living model. At least one extinct hominin subclade, Paranthropus, has a pattern of dental life history related variables that most likely set it apart from the life histories of both modern humans and chimpanzees. PMID- 18380865 TI - Cranial base evolution within the hominin clade. AB - The base of the cranium (i.e. the basioccipital, the sphenoid and the temporal bones) is of particular interest because it undergoes significant morphological change within the hominin clade, and because basicranial morphology features in several hominin species diagnoses. We use a parsimony analysis of published cranial and dental data to predict the cranial base morphology expected in the hypothetical last common ancestor of the Pan-Homo clade. We also predict the primitive condition of the cranial base for the hominin clade, and document the evolution of the cranial base within the major subclades within the hominin clade. This analysis suggests that cranial base morphology has continued to evolve in the hominin clade, both before and after the emergence of the genus Homo. PMID- 18380866 TI - The facial skeleton of the chimpanzee-human last common ancestor. AB - This review uses the current morphological evidence to evaluate the facial morphology of the hypothetical last common ancestor (LCA) of the chimpanzee/bonobo (panin) and human (hominin) lineages. Some of the problems involved in reconstructing ancestral morphologies so close to the formation of a lineage are discussed. These include the prevalence of homoplasy and poor phylogenetic resolution due to a lack of defining derived features. Consequently the list of hypothetical features expected in the face of the LCA is very limited beyond its hypothesized similarity to extant Pan. It is not possible to determine with any confidence whether the facial morphology of any of the current candidate LCA taxa (Ardipithecus kadabba, Ardipithecus ramidus, Orrorin tugenensis and Sahelanthropus tchadensis) is representative of the LCA, or a stem hominin, or a stem panin or, in some cases, a hominid predating the emergence of the hominin lineage. The major evolutionary trends in the hominin lineage subsequent to the LCA are discussed in relation to the dental arcade and dentition, subnasal morphology and the size, position and prognathism of the facial skeleton. PMID- 18380864 TI - A natural history of the human mind: tracing evolutionary changes in brain and cognition. AB - Since the last common ancestor shared by modern humans, chimpanzees and bonobos, the lineage leading to Homo sapiens has undergone a substantial change in brain size and organization. As a result, modern humans display striking differences from the living apes in the realm of cognition and linguistic expression. In this article, we review the evolutionary changes that occurred in the descent of Homo sapiens by reconstructing the neural and cognitive traits that would have characterized the last common ancestor and comparing these with the modern human condition. The last common ancestor can be reconstructed to have had a brain of approximately 300-400 g that displayed several unique phylogenetic specializations of development, anatomical organization, and biochemical function. These neuroanatomical substrates contributed to the enhancement of behavioral flexibility and social cognition. With this evolutionary history as precursor, the modern human mind may be conceived as a mosaic of traits inherited from a common ancestry with our close relatives, along with the addition of evolutionary specializations within particular domains. These modern human specific cognitive and linguistic adaptations appear to be correlated with enlargement of the neocortex and related structures. Accompanying this general neocortical expansion, certain higher-order unimodal and multimodal cortical areas have grown disproportionately relative to primary cortical areas. Anatomical and molecular changes have also been identified that might relate to the greater metabolic demand and enhanced synaptic plasticity of modern human brain's. Finally, the unique brain growth trajectory of modern humans has made a significant contribution to our species' cognitive and linguistic abilities. PMID- 18380867 TI - Inferences regarding the diet of extinct hominins: structural and functional trends in dental and mandibular morphology within the hominin clade. AB - This contribution investigates the evolution of diet in the Pan-Homo and hominin clades. It does this by focusing on 12 variables (nine dental and three mandibular) for which data are available about extant chimpanzees, modern humans and most extinct hominins. Previous analyses of this type have approached the interpretation of dental and gnathic function by focusing on the identification of the food consumed (i.e. fruits, leaves, etc.) rather than on the physical properties (i.e. hardness, toughness, etc.) of those foods, and they have not specifically addressed the role that the physical properties of foods play in determining dental adaptations. We take the available evidence for the 12 variables, and set out what the expression of each of those variables is in extant chimpanzees, the earliest hominins, archaic hominins, megadont archaic hominins, and an inclusive grouping made up of transitional hominins and pre modern Homo. We then present hypotheses about what the states of these variables would be in the last common ancestor of the Pan-Homo clade and in the stem hominin. We review the physical properties of food and suggest how these physical properties can be used to investigate the functional morphology of the dentition. We show what aspects of anterior tooth morphology are critical for food preparation (e.g. peeling fruit) prior to its ingestion, which features of the postcanine dentition (e.g. overall and relative size of the crowns) are related to the reduction in the particle size of food, and how information about the macrostructure (e.g. enamel thickness) and microstructure (e.g. extent and location of enamel prism decussation) of the enamel cap might be used to make predictions about the types of foods consumed by extinct hominins. Specifically, we show how thick enamel can protect against the generation and propagation of cracks in the enamel that begin at the enamel-dentine junction and move towards the outer enamel surface. PMID- 18380870 TI - 25 years of published and unpublished coagulation-therapy data for Haemate P. PMID- 18380869 TI - The evolutionary history of the hominin hand since the last common ancestor of Pan and Homo. AB - Molecular evidence indicates that the last common ancestor of the genus Pan and the hominin clade existed between 8 and 4 million years ago (Ma). The current fossil record indicates the Pan-Homo last common ancestor existed at least 5 Ma and most likely between 6 and 7 Ma. Together, the molecular and fossil evidence has important consequences for interpreting the evolutionary history of the hand within the tribe Hominini (hominins). Firstly, parsimony supports the hypothesis that the hand of the last common ancestor most likely resembled that of an extant great ape overall (Pan, Gorilla, and Pongo), and that of an African ape in particular. Second, it provides a context for interpreting the derived changes to the hand that have evolved in various hominins. For example, the Australopithecus afarensis hand is likely derived in comparison with that of the Pan-Homo last common ancestor in having shorter fingers relative to thumb length and more proximo-distally oriented joints between its capitate, second metacarpal, and trapezium. This evidence suggests that these derived features evolved prior to the intensification of stone tool-related hominin behaviors beginning around 2.5 Ma. However, a majority of primitive features most likely present in the Pan-Homo last common ancestor are retained in the hands of Australopithecus, Paranthropus/early Homo, and Homo floresiensis. This evidence suggests that further derived changes to the hands of other hominins such as modern humans and Neandertals did not evolve until after 2.5 Ma and possibly even later than 1.5 Ma, which is currently the earliest evidence of Acheulian technology. The derived hands of modern humans and Neandertals may indicate a morphological commitment to tool-related manipulative behaviors beyond that observed in other hominins, including those (e.g. H. floresiensis) which may be descended from earlier tool making species. PMID- 18380871 TI - A systematic overview of the first pasteurised VWF/FVIII medicinal product, Haemate P/ Humate -P: history and clinical performance. AB - Patients with von Willebrand disease (VWD) and haemophilia A (HA) lack, to varying degrees, the von Willebrand factor (VWF) and coagulation factor VIII (FVIII) that are critical for normal haemostasis. These conditions in turn make patients prone to uncontrolled bleeding. Historically, patients with severe forms of VWD or HA were crippled before adulthood and their life expectancy was significantly reduced. Over the past decades, specific coagulation factor replacement therapies including Haemate P, have been developed to help patients achieve and maintain normal haemostasis. Haemate P is a human, plasma-derived VWF/FVIII medicinal product, which was first licensed in Germany in 1981 for the treatment of HA-associated bleeding. It has since then come to be accepted as the gold standard for both the treatment and prophylaxis of bleeding in VWD, especially in cases where desmopressin [1-deamino-8-D-arginine vasopressin (DDAVP)] has been ineffective. Haemate P was the first effectively virus inactivated (pasteurisation: 60 degrees C for 10 h in aqueous solution) FVIII product, whereby the risk of potentially threatening infective complications of plasma-derived products was reduced. Haemate P was also shown to have a VWF multimer profile remarkably close to that of normal plasma. This bibliographic review presents previously unpublished clinical data of Haemate P, based upon internal clinical study reports of the proprietor, CSL Behring, in addition to data already presented in other publications. The data demonstrate a predictable and well-characterised pharmacokinetic profile, and a proven record of short- and long-term safety, while effectively correcting the haemostatic defects in VWD and HA. Recently available data have also shown Haemate P to be of haemostatic value in exceptional clinical circumstances including surgical interventions. By virtue of its plasma-derived combination of VWF and FVIII, in addition to its high VWF:FVIII content ratio (2.4:1), Haemate P is also associated with successful immune tolerance induction in those patients developing inhibitor antibodies. Although the theoretical risk of thromboembolic complications does exist while receiving Haemate P, as it does with any FVIII replacement therapy, the incidence of such complications has remained notably low. Given the robust data that have accumulated for the use of Haemate P, dosing recommendations are also described in this review; the recommendations are tailored to patient-specific contexts including baseline VWF and FVIII levels in plasma and the type of surgical intervention being undertaken. A wide variety of studies have also provided data on paediatric and geriatric populations, all of which have suggested that Haemate P can be safely and effectively used in a wide variety of clinical circumstances. PMID- 18380868 TI - Locomotion and posture from the common hominoid ancestor to fully modern hominins, with special reference to the last common panin/hominin ancestor. AB - Based on our knowledge of locomotor biomechanics and ecology we predict the locomotion and posture of the last common ancestors of (a) great and lesser apes and their close fossil relatives (hominoids); (b) chimpanzees, bonobos and modern humans (hominines); and (c) modern humans and their fossil relatives (hominins). We evaluate our propositions against the fossil record in the context of a broader review of evolution of the locomotor system from the earliest hominoids of modern aspect (crown hominoids) to early modern Homo sapiens. While some early East African stem hominoids were pronograde, it appears that the adaptations which best characterize the crown hominoids are orthogrady and an ability to abduct the arm above the shoulder - rather than, as is often thought, manual suspension sensu stricto. At 7-9 Ma (not much earlier than the likely 4-8 Ma divergence date for panins and hominins, see Bradley, 2008) there were crown hominoids in southern Europe which were adapted to moving in an orthograde posture, supported primarily on the hindlimb, in an arboreal, and possibly for Oreopithecus, a terrestrial context. By 7 Ma, Sahelanthropus provides evidence of a Central African hominin, panin or possibly gorilline adapted to orthogrady, and both orthogrady and habitually highly extended postures of the hip are evident in the arboreal East African protohominin Orrorin at 6 Ma. If the traditional idea that hominins passed through a terrestrial 'knuckle-walking' phase is correct, not only does it have to be explained how a quadrupedal gait typified by flexed postures of the hindlimb could have preadapted the body for the hominin acquisition of straight-legged erect bipedality, but we would have to accept a transition from stem-hominoid pronogrady to crown hominoid orthogrady, back again to pronogrady in the African apes and then back to orthogrady in hominins. Hand assisted arboreal bipedality, which is part of a continuum of orthograde behaviours, is used by modern orangutans to forage among the small branches at the periphery of trees where the core hominoid dietary resource, ripe fruit, is most often to be found. Derivation of habitual terrestrial bipedality from arboreal hand-assisted bipedality requires fewer transitions, and is also kinematically and kinetically more parsimonious. PMID- 18380872 TI - Growth of synthetic stromatolites and wrinkle structures in the absence of microbes - implications for the early fossil record. AB - Stromatolites and wrinkle structures are often taken to be an important indicator for early life. While both may be shaped by microbial mat growth, this can be open to doubt, so that the contribution of abiotic processes in their construction always needs to be established (Grotzinger & Knoll, 1999). We here report laboratory spray deposition experiments that can generate stromatolites and wrinkle structures in the absence of microbes. These minicolumnar and sometimes branched stromatolites are produced artificially by the aggregation of a synthetic colloid in a turbulent flow regime. They self-organize at the relatively low particle concentrations found in the outer parts of a spray beam. This contrasts with adjacent stratiform deposits that are produced by high rates of colloid deposition and relatively low sediment viscosities found in the centre of a spray beam. These stratiform laminae become subsequently wrinkled during hardening of the colloid. These results support numerical models that together suggest that physicochemical processes are capable of generating laminated sedimentary structures without the direct participation of biology. Geological environments where comparable abiogenic stromatolites and wrinkle structures may be found include: splash-zone silica sinters, desert varnish crusts and early Archean cherts formed from silica gel precursors. PMID- 18380873 TI - Seawater Mg/Ca controls polymorph mineralogy of microbial CaCO3: a potential proxy for calcite-aragonite seas in Precambrian time. AB - A previously published hydrothermal brine-river water mixing model driven by ocean crust production suggests that the molar Mg/Ca ratio of seawater (mMg/Ca(sw)) has varied significantly (approximately 1.0-5.2) over Precambrian time, resulting in six intervals of aragonite-favouring seas (mMg/Ca(sw) > 2) and five intervals of calcite-favouring seas (mMg/Ca(sw) < 2) since the Late Archaean. To evaluate the viability of microbial carbonates as mineralogical proxy for Precambrian calcite-aragonite seas, calcifying microbial marine biofilms were cultured in experimental seawaters formulated over the range of Mg/Ca ratios believed to have characterized Precambrian seawater. Biofilms cultured in experimental aragonite seawater (mMg/Ca(sw) = 5.2) precipitated primarily aragonite with lesser amounts of high-Mg calcite (mMg/Ca(calcite) = 0.16), while biofilms cultured in experimental calcite seawater (mMg/Ca(sw) = 1.5) precipitated exclusively lower magnesian calcite (mMg/Ca(calcite) = 0.06). Furthermore, Mg/Ca(calcite )varied proportionally with Mg/Ca(sw). This nearly abiotic mineralogical response of the biofilm CaCO3 to altered Mg/Ca(sw) is consistent with the assertion that biofilm calcification proceeds more through the elevation of , via metabolic removal of CO2 and/or H+, than through the elevation of Ca2+, which would alter the Mg/Ca ratio of the biofilm's calcifying fluid causing its pattern of CaCO3 polymorph precipitation (aragonite vs. calcite; Mg-incorporation in calcite) to deviate from that of abiotic calcification. If previous assertions are correct that the physicochemical properties of Precambrian seawater were such that Mg/Ca(sw) was the primary variable influencing CaCO3 polymorph mineralogy, then the observed response of the biofilms' CaCO3 polymorph mineralogy to variations in Mg/Ca(sw), combined with the ubiquity of such microbial carbonates in Precambrian strata, suggests that the original polymorph mineralogy and Mg/Ca(calcite )of well-preserved microbial carbonates may be an archive of calcite-aragonite seas throughout Precambrian time. These results invite a systematic evaluation of microbial carbonate primary mineralogy to empirically constrain Precambrian seawater Mg/Ca. PMID- 18380874 TI - Earliest fossil record of bacterial-cyanobacterial mat consortia: the early Silurian Passage Creek biota (440 Ma, Virginia, USA). AB - Cyanobacteria in terrestrial and aquatic habitats are frequently associated with heterotrophic bacteria, and such associations are most often metabolically interactive. Functionally, the members of such bacterial-cyanobacterial consortia benefit from diverse metabolic capabilities of their associates, thus exceeding the sum of their parts. Such associations may have been just as ubiquitous in the past, but the fossil record has not produced any direct evidence for such associations to date. In this paper, we document fossil bacteria associated with a macrophytic cyanobacterial mat in the early Silurian (Llandovery) Massanutten Sandstone of Virginia, USA. Both the bacterial and the cyanobacterial cells are preserved by mineral replacement (pyrite subsequently replaced by iron oxyhydroxides) within an amorphous carbonaceous matrix which represents the common exopolysaccharide investment of the cyanobacterial colony. The bacteria are rod-shaped, over 370 nm long and 100 nm in diameter, and occur both as isolated cells and as short filaments. This occurrence represents the oldest fossil evidence for bacterial-cyanobacterial associations, documenting that such consortia were present 440 Ma ago, and revealing the potential for them to be recognized deeper in the fossil record. PMID- 18380875 TI - Putative cryptoendolithic life in Devonian pillow basalt, Rheinisches Schiefergebirge, Germany. AB - Middle Devonian (Givetian) pillow basalt and inter-pillow breccia from the Rheinisches Schiefergebirge in Germany were found to contain putative biogenic filaments that indicate that life once proliferated within these volcanic rocks. Mineralized filaments are found in carbonate amygdules (vesicles filled by carbonate cement) in the volcanic rock, where they started to form on the internal surface of the once water-filled vesicles. Biogenicity of the filaments is indicated by (1) their size and shape resembling modern microorganisms including a constant diameter along the length of curved filaments, (2) their independence of crystal faces or cleavage planes, (3) branching patterns reminiscent of modern microorganisms, and (4) their spatial clustering and preferential occurrence close to the margin of pillows and in the inter-pillow breccias. A time lag between the deposition of pillow basalt and the activity of endoliths is revealed by the sequence of carbonate cements filling the amygdules. The putative filamentous microorganisms thrived after the formation of early fibrous rim cement, but before later equant calcite spar filled most of the remaining porosity. Microbial clay authigenesis analogous to the encrustation of prokaryotes in modern iron-rich environments led to the preservation of filaments. The filaments predominantly consist of the clay minerals chamosite and illite. Having dwelled in water-filled vesicles, the Devonian basalt-hosted filaments apparently represent cryptoendoliths. This finding suggests that a previously unrecognized niche for life exists within volcanic rock. PMID- 18380876 TI - Early colonization of thermal niches in a silica-depositing hot spring in central Tibet. AB - Thermophilic microbial mats dominated by the anoxygenic phototroph Roseiflexus castenholzii commonly develop around sinter-depositing geysers in the Daggyai Tso geothermal field of central Tibet. In this study we used morphological and molecular genetic techniques to reveal a diverse pioneer biofilm community including both archaea and bacteria involved in early colonization of such thermal niches at temperatures ranging from 46 to 77 degrees C. Sinter precipitation and biomineralization were evident at all locations, but the latter was selective between taxa and most evident on filamentous cells. Evidence for possible indirect biosignatures from biofilms overwhelmed by sinter deposition was found. Succession to a mature community appeared to relate to the growth rate for key taxa outpacing that of silicification within an optimum temperature range of 54-61 degrees C. The thin surface layer of silicification-resistant cyanobacteria that developed on the surface of mature mats may play a role in preventing biomineralization of the susceptible R. castenholzii beneath within these communities. PMID- 18380877 TI - Characterization of extracellular minerals produced during dissimilatory Fe(III) and U(VI) reduction at 100 degrees C by Pyrobaculum islandicum. AB - In order to gain insight into the significance of biotic metal reduction and mineral formation in hyperthermophilic environments, metal mineralization as a result of the dissimilatory reduction of poorly crystalline Fe(III) oxide, and U(VI) reduction at 100 degrees C by Pyrobaculum islandicum was investigated. When P. islandicum was grown in a medium with poorly crystalline Fe(III) oxide as an electron acceptor and hydrogen as an electron donor, the Fe(III) oxide was reduced to an extracellular, ultrafine-grained magnetite with characteristics similar to that found in some hot environments and that was previously thought to be of abiotic origin. Furthermore, cell suspensions of P. islandicum rapidly reduced the soluble and oxidized form of uranium, U(VI), to extracellular precipitates of the highly insoluble U(IV) mineral, uraninite (UO(2)). The reduction of U(VI) was dependent on the presence of hydrogen as the electron donor. These findings suggest that microbes may play a key role in metal deposition in hyperthermophilic environments and provide a plausible explanation for such phenomena as magnetite accumulation and formation of uranium deposits at ca. 100 degrees C. PMID- 18380878 TI - Fossilized microorganisms associated with zeolite-carbonate interfaces in sub seafloor hydrothermal environments. AB - In this paper we describe carbon-rich filamentous structures observed in association with the zeolite mineral phillipsite from sub-seafloor samples drilled and collected during the Ocean Drilling Program (ODP) Leg 197 at the Emperor Seamounts. The filamentous structures are approximately 5 microm thick and approximately 100-200 microm in length. They are found attached to phillipsite surfaces in veins and entombed in vein-filling carbonates. The carbon content of the filaments ranges between approximately 10 wt% C and 55 wt% C. They further bind to propidium iodide (PI), which is a dye that binds to damaged cell membranes and remnants of DNA. Carbon-rich globular microstructures, 1-2 microm in diameter, are also found associated with the phillipsite surfaces as well as within wedge-shaped cavities in phillipsite assemblages. The globules have a carbon content that range between approximately 5 wt% C and 55 wt% C and they bind to PI. Ordinary globular iron oxides found throughout the samples differ in that they contain no carbon and do not bind to the dye PI. The carbon-rich globules are mostly concentrated to a film-like structure that is attached to the phillipsite surfaces. This film has a carbon content that ranges between approximately 25 wt% C and 75 wt% C and partially binds to PI. EDS analyses show that the carbon in all structures described are not associated with calcium and therefore not bound in carbonates. The carbon content and the binding to PI may indicate that the filamentous structures could represent fossilized filamentous microorganisms, the globules could represent fossilized microbial cells and the film-like structures could represent a microbially produced biofilm. Our results extend the knowledge of possible habitable niches for a deep biosphere in sub seafloor environments and suggests, as phillipsite is one of the most common zeolite mineral in volcanic rocks of the oceanic crust, that it could be a common feature in the oceanic crust elsewhere. PMID- 18380879 TI - Carbon pools and isotopic trends in a hypersaline cyanobacterial mat. AB - The fine-scale depth distribution of major carbon pools and their stable carbon isotopic signatures (delta(13)C) were determined in a cyanobacterial mat (Salin de-Giraud, Camargue, France) to study early diagenetic alterations and the carbon preservation potential in hypersaline mat ecosystems. Particular emphasis was placed on the geochemical role of extracellular polymeric substances (EPS). Total carbon (C(tot)), organic carbon (C(org)), total nitrogen (N(tot)), total hydrolysable amino acids (THAA), carbohydrates, cyanobacteria-derived hydrocarbons (8-methylhexadecane, n-heptadec-5-ene, n-heptadecane) and EPS showed highest concentrations in the top millimetre of the mat and decreased with depth. The hydrocarbons attributed to cyanobacteria showed the strongest decrease in concentration with depth. This correlated well with the depth profiles of oxygenic photosynthesis and oxygen, which were detected in the top 0.6 and 1.05 mm, respectively, at a high down-welling irradiance (1441 micromol photons m(-2) s(-1)). At depths beneath the surface layer, the C(org) was composed mainly of amino acids and carbohydrates. A resistance towards microbial degradation could have resulted from interactions with diverse functional groups present in biopolymers (EPS) and with minerals deposited in the mat. A (13)C enrichment with depth for the total carbon pool (C(tot)) was observed, with delta(13)C values ranging from -16.3 per thousand at the surface to -11.3 per thousand at 9-10 mm depth. Total lipids depicted a delta(13)C value of -17.2 per thousand in the top millimetre and then became depleted in (13)C with depth (-21.7 to -23.3 per thousand). The delta(13)C value of EPS varied only slightly with depth (-16.1 to 17.3 per thousand) and closely followed the delta(13)C value of C(org) at depths beneath 4 mm. The EPS represents an organic carbon pool of preservation potential during early stages of diagenesis in recent cyanobacterial mats as a result of a variety of possible interactions. Their analyses might improve our understanding of fossilized microbial remains from mat ecosystems. PMID- 18380881 TI - Microbial interactions with physical sediment dynamics, and their significance for the interpretation of Earth's biological history. PMID- 18380882 TI - An actualistic perspective into Archean worlds - (cyano-)bacterially induced sedimentary structures in the siliciclastic Nhlazatse Section, 2.9 Ga Pongola Supergroup, South Africa. AB - Extensive microbial mats colonize sandy tidal flats that form along the coasts of today's Earth. The microbenthos (mainly cyanobacteria) respond to the prevailing physical sediment dynamics by biostabilization, baffling and trapping, as well as binding. This biotic-physical interaction gives rise to characteristic microbially induced sedimentary structures (MISS) that differ greatly from both purely physical structures and from stromatolites. Actualistic studies of the MISS on modern tidal flats have been shown to be the key for understanding equivalent fossil structures that occur in tidal and shelf sandstones of all Earth ages. However, until now the fossil record of Archean MISS has been poor, and relatively few specimens have been found. This paper describes a study location that displays a unique assemblage with a multitude of exceptionally preserved MISS in the 2.9-Ga-old Pongola Supergroup, South Africa. The 'Nhlazatse Section' includes structures such as 'erosional remnants and pockets', 'multidirected ripple marks', 'polygonal oscillation cracks', and 'gas domes'. Optical and geochemical analyses support the biogenicity of microscopic textures such as filamentous laminae or 'orientated grains'. Textures resembling filaments are lined by iron oxide and hydroxides, as well as clay minerals. They contain organic matter, whose isotope composition is consistent with carbon of biological origin. The ancient tidal flats of the Nhlazatse Section record four microbial mat facies that occur in modern tidal settings as well. We distinguish endobenthic and epibenthic microbial mats, including planar, tufted, and spongy subtypes. Each microbial mat facies is characterized by a distinct set of MISS, and relates to a typical tidal zone. The microbial mat structures are preserved in situ, and are consistent with similar features constructed today by benthic cyanobacteria. However, other mat-constructing microorganisms also could have formed the structures in the Archean tidal flats. PMID- 18380883 TI - Wave and sediment dynamics along a shallow subtidal sandy beach inhabited by modern stromatolites. AB - To help define the habitat of modern marine stromatolites, wave-dominated flow and sediment transport were studied in the shallow subtidal region (1-2 m depth) along the slightly concave, windward face of Highborne Cay, Exuma, Bahamas - the only face of the cay that includes a population of stromatolites concentrated near the region of highest curvature of the beach. Wave energy impacting this island's most exposed beach was driven by local wind forcing which increases largely in response to the passage of atmospheric disturbances that typically affect the region for periods of a few days. Although some wave energy is almost always noted (maximum horizontal orbital speeds at the bottom are rarely <10 cm s(-1)), wave conditions remain comparatively calm until local winds increase above speeds of approximately 3-4 m s(-1) at which point maximum wave speeds rapidly increase to 50-80 cm s(-1). Stromatolites, which are largely restricted to the shoreward side of a shallow platform reef, are sheltered by the reef beyond which wave speeds are one to four times higher (depending on tidal stage). Moreover, stromatolite populations are predominantly found along a region of this wave-exposed beach that experiences comparatively reduced wave energy because of the curved morphology of the island's face. Maximum wave speeds are 1.4 to 2 times higher along more northern sections of the beach just beyond the locus of stromatolite populations. A quantitative model of sediment transport was developed that accurately predicted accumulation of suspended sediment in sediment traps deployed in the shallow subtidal zone along this beach. This model, coupled with in situ wave records, indicates that gross rates of suspended sediment deposition should be two to three times higher northward of the main stromatolite populations. Regions of the beach containing stromatolites nevertheless should experience significant rates of gross suspended sediment deposition averaging 7-10 g cm(-2) day(-1) ( approximately 4-6 cm day(-1)). Results suggest that one axis of the habitat of modern marine stromatolites may be defined by a comparatively narrow range of flow energy and sediment transport conditions. PMID- 18380884 TI - Stromatolite branching in the Neoproterozoic of the Centralian Superbasin, Australia: an investigation into sedimentary and microbial control of stromatolite morphology. AB - The extensive and well-preserved Neoproterozoic Acaciella australica Stromatolite assemblage of Australia is ideal for examining the relative roles of microbial and environmental influences on stromatolite branching and stromatolite macrostructure across a wide geographical area. Detailed sedimentological analyses indicate that the basal hemispheroidal section of bioherms contains abundant sediment. By contrast, the columnar sections of bioherms are composed almost exclusively of micritic laminae. These micritic laminae display little evidence for environmental, especially sedimentary, control over stromatolite morphology. The change from a hemispheroidal morphology to branching morphology is linked to variations in the relative contributions of sediment and framework growth. The shift to columns appears to be closely linked to a decrease in sediment supply that resulted in a more stable environment in which microbially mediated framework growth began to control stromatolite morphology. Branching in the A. australica assemblage stromatolites appears to be caused by shifting sedimentary and microbial control on stromatolite morphology. PMID- 18380885 TI - Calcium carbonate precipitation in cyanobacterial mats from sandy tidal flats of the North Sea. AB - Precipitated calcium carbonate was found in annual cyanobacterial mats developing on the beaches of the North Sea barrier island Schiermonnikoog (the Netherlands). A variety of different calcium carbonate morphs were found in the cyanobacterial mucous secretions and identified by light- and scanning electron microscopy and X ray diffraction. Most of the calcium carbonate seemed to be associated with degrading extracellular polymer. It is conceived that supersaturation of calcium carbonate resulted from the periodic evaporation of the mats and from the release of calcium from the cyanobacterial mucous as a result of its degradation. The analysis of the carbon stable isotopic composition of the calcium carbonate showed only a slight depletion of (13)C, indicating that it did not in major part originated from the decomposition of organic matter. The delta(18)O values of the calcium carbonate confirmed the temperature differences between spring and summer but excluded the possibility that excessive evaporative events controlled precipitation. The precipitation of calcium carbonate could represent a potential factor enhancing the stabilization of intertidal siliciclastic sediments through cementing the sand. The discovery of massive calcium carbonate precipitation in these cyanobacterial mats was unexpected and hitherto unknown. PMID- 18380886 TI - Microbial stabilization of riverine sediments by extracellular polymeric substances. AB - Sediment stability is a critical component for the understanding of cohesive sediment dynamics. Traditionally, physico-chemical sediment conditions have been regarded as most important drivers of sediment stability. However, over the last decade, the stabilization of sediment by biological activity, particularly the influence of highly hydrated matrices of extracellular polymeric substances (EPS) has been given increasing attention. However, most studies have focused on the sediment/water interface and, usually, of marine systems. The present study exploits current knowledge of EPS dynamics from marine systems and applies it to freshwater habitats, also considering a wide range of biological and physico chemical variables. Natural sediments were taken from a freshwater site with high levels of heavy metal pollution (Lauffen reservoir, River Neckar, Germany). Vertical profiles from the flocculent surface layer to depth of 50 cm within the sediment were investigated, monthly, over the course of year. Tubificidae and Chironomidae larvae constituted the majority of the macrofauna. Despite the turbidity of the water column, a highly diverse and abundant microphytobenthic community of diatoms (11-82 microg g(-1) DW) was found at the sediment surface closely associated with high numbers of bacteria (10(9) cells g(-1) DW). The concentrations of all EPS moieties were remarkably high (0.1-0.5, 1.7-3.8, 0.9 5.2 mg g(-1) DW, for colloidal and bound carbohydrates and proteins, respectively) and levels were comparable to those determined in intertidal studies. The microalgal and bacterial biomass both showed strong correlations with the colloidal and bound EPS carbohydrate fractions. The data suggested that the present macrofauna as well as the metabolic activities of microalgae and bacteria interact with sedimentological factors to influence the properties of the sediment by binding fine-grained sediment, changing water content and enhancing the organic content through secretion products. The colloidal and bound EPS moieties showed strong correlation with the critical shear stress for erosion over sediment depth. It is suggested that the cohesive strength of the sediment was controlled by a high number of active adsorption sites and higher charge densities in fine grained sediments. The EPS network may significantly enhance this by embedding particles and permeating the void space but also in offering additional ionic binding sites and cross-linkages. PMID- 18380887 TI - Microalgal mediation of ripple mobility. AB - The interaction between physical and biological factors responsible for the cessation of ripple migration on a sandy intertidal flat was examined during a microalgal bloom period in late winter/early spring, as part of a wider study into the biostabilisation of intertidal sediments. Ripple positions and ripple geometry were monitored, and surface sediment was sampled, at weekly intervals over a 5-week period. Ripples remained in the same position for at least 4 weeks, during which time there was a progressive reduction in bedform height (smoothing) and deposition of some 1.5 cm sediment, mainly in the ripple troughs (surface levelling). The mean chlorophyll a (chl a) sediment content was 6.0 microg gDW( 1) (DW: dry weight) (0-1 mm depth fraction), with a maximum value of 7.4 microg gDW(-1) half way through the bloom. Mean colloidal-S carbohydrate (S: saline extraction) content was 131 microg GE gDW(-1) (GE: glucose equivalent) (0-1 mm), with a maximum of 261 microg GE gDW(-1 )towards the end of the bloom. Important accessory pigments were peridinin (indicative of dinophytes) and fucoxanthin (diatoms). Stepwise multiple regression showed that peridinin was the best predictor of chl a. For the first time, in situ evidence for the mediation of (wave) ripple migration by microalgae is provided. Results indicate that diatoms, and quite possibly dinophytes, can have a significant effect on intertidal flat ripple mobility on a temporal scale of weeks. In addition, microalgal effects appear capable of effecting a reduction in bed roughness on a spatial scale of up to 10(-2 )m, with a subsequent reduction in bottom stress and bed erodability. It is suggested that a unique combination of environmental conditions, in conjunction with the microalgal bloom(s), promoted the initial cessation of ripple movement, and that stationary-phase, diatom-derived extracellular polymeric substances (EPS) (and possibly dinophyte-derived EPS) may have prolonged the condition. It is reasonable to suppose that ripple stabilisation by similar processes may have contributed to ripple mark preservation in the geological record. A conceptual model of sandy intertidal flat processes is presented, illustrating two conditions: (i) a low EPS/microalgae sediment content with low ripple stabilisation and preservation potential; and (ii) a high EPS/microalgae content with higher preservation potential. PMID- 18380888 TI - Tube structures of probable microbial origin in the Neoarchean Carawine Dolomite, Hamersley Basin, Western Australia. AB - The approximately 2.63 Ga Carawine Dolomite, Hamersley Basin, Western Australia, preserves tube structures of probable microbial origin that formed in a low energy environment. The tubes are 0.4-1.8 cm in diameter and at least 10-16 cm long in outcrop. The tubes are defined by dark, 45-microm-thick dolomicritic walls, whereas the tube fill and host rock are composed of 30 microm, cloudy dolomite crystals and rare 170- to 425-microm-wide, dark well-sorted clasts. Closely spaced, rarely discontinuous laminae coat the insides of tubes; less closely spaced, peaked, discontinuous laminae coat the outsides of tubes. The laminae on the outsides of tubes are often intercalated with mammilate structures. The presence of probable microbial coatings on both the insides and the outsides of the tube walls requires that the tubes formed above the sediment water interface. These tube structures probably formed during gas-charged fluid escape, similar to tubes observed in ancient and modern hydrocarbon seeps and cylindrical water transfer structures in sandstones. The laminae that coat the tubes have very similar geometries to modern biofilms that form in both turbulent and laminar flow, and their geometries probably reflect flow conditions during the fluid escape. The identification of these structures suggests that the preserved interaction between fluid escape and microbial growth in carbonates may be more common than previously thought. PMID- 18380889 TI - Primary breast lymphoma: patient profile, outcome and prognostic factors. A multicentre Rare Cancer Network study. AB - BACKGROUND: To asses the clinical profile, treatment outcome and prognostic factors in primary breast lymphoma (PBL). METHODS: Between 1970 and 2000, 84 consecutive patients with PBL were treated in 20 institutions of the Rare Cancer Network. Forty-six patients had Ann Arbor stage IE, 33 stage IIE, 1 stage IIIE, 2 stage IVE and 2 an unknown stage. Twenty-one underwent a mastectomy, 39 conservative surgery and 23 biopsy; 51 received radiotherapy (RT) with (n = 37) or without (n = 14) chemotherapy. Median RT dose was 40 Gy (range 12-55 Gy). RESULTS: Ten (12%) patients progressed locally and 43 (55%) had a systemic relapse. Central nervous system (CNS) was the site of relapse in 12 (14%) cases. The 5-yr overall survival, lymphoma-specific survival, disease-free survival and local control rates were 53%, 59%, 41% and 87% respectively. In the univariate analyses, favorable prognostic factors were early stage, conservative surgery, RT administration and combined modality treatment. Multivariate analysis showed that early stage and the use of RT were favorable prognostic factors. CONCLUSION: The outcome of PBL is fair. Local control is excellent with RT or combined modality treatment but systemic relapses, including that in the CNS, occurs frequently. PMID- 18380891 TI - Epidermal growth factor receptor levels are reduced in mice with targeted disruption of the protein kinase A catalytic subunit. AB - BACKGROUND: Epidermal Growth Factor Receptor (EGFR) is a key target molecule in current treatment of several neoplastic diseases. Hence, in order to develop and improve current drugs targeting EGFR signalling, an accurate understanding of how this signalling pathway is regulated is required. It has recently been demonstrated that inhibition of cAMP-dependent protein kinase (PKA) induces a ligand-independent internalization of EGFR. Cyclic-AMP-dependent protein kinase consists of a regulatory dimer bound to two catalytic subunits. RESULTS: We have investigated the effect on EGFR levels after ablating the two catalytic subunits, Calpha and Cbeta in two different models. The first model used targeted disruption of either Calpha or Cbeta in mice whereas the second model used Calpha and Cbeta RNA interference in HeLa cells. In both models we observed a significant reduction of EGFR expression at the protein but not mRNA level. CONCLUSION: Our results suggest that PKA may represent a target that when manipulated can maintain EGFR protein levels at the single cell level as well as in intact animals. PMID- 18380890 TI - Wheel running from a juvenile age delays onset of specific motor deficits but does not alter protein aggregate density in a mouse model of Huntington's disease. AB - BACKGROUND: Huntington's disease (HD) is a neurodegenerative disorder predominantly affecting the cerebral cortex and striatum. Transgenic mice (R6/1 line), expressing a CAG repeat encoding an expanded polyglutamine tract in the N terminus of the huntingtin protein, closely model HD. We have previously shown that environmental enrichment of these HD mice delays the onset of motor deficits. Furthermore, wheel running initiated in adulthood ameliorates the rear paw clasping motor sign, but not an accelerating rotarod deficit. RESULTS: We have now examined the effects of enhanced physical activity via wheel running, commenced at a juvenile age (4 weeks), with respect to the onset of various behavioral deficits and their neuropathological correlates in R6/1 HD mice. HD mice housed post-weaning with running wheels only, to enhance voluntary physical exercise, have delayed onset of a motor co-ordination deficit on the static horizontal rod, as well as rear-paw clasping, although the accelerating rotarod deficit remains unaffected. Both wheel running and environmental enrichment rescued HD-induced abnormal habituation of locomotor activity and exploratory behavior in the open field. We have found that neither environment enrichment nor wheel running ameliorates the shrinkage of the striatum and anterior cingulate cortex (ACC) in HD mice, nor the overall decrease in brain weight, measured at 9 months of age. At this age, the density of ubiquitinated protein aggregates in the striatum and ACC is also not significantly ameliorated by environmental enrichment or wheel running. CONCLUSION: These results indicate that enhanced voluntary physical activity, commenced at an early presymptomatic stage, contributes to the positive effects of environmental enrichment. However, sensory and cognitive stimulation, as well as motor stimulation not associated with running, may constitute major components of the therapeutic benefits associated with enrichment. Comparison of different environmental manipulations, performed in specific time windows, can identify critical periods for the induction of neuroprotective 'brain reserve' in animal models of HD and related neurodegenerative diseases. PMID- 18380892 TI - Effects of electromagnetic fields of low frequency and low intensity on rat metabolism. AB - A series of experiments on rats have been performed, to study the effects of long time (50 days) exposure to electromagnetic fields of extremely low frequency (ELF, i.e. less than 100 Hz) and amplitude (non thermal), testing whether the metabolic processes would be affected. The background lies on recent observations on the behaviour of isolated enzymes in vitro exposed to EFL fields. In these experiments, the cyclotron (or Larmor) frequency of the metallic ion has been used to "stimulate" the metalloproteins redox-active site, thus obtaining a clear variation of the enzyme functionality. In this paper we have extended for the first time the check to more complex animal metabolism. The novelty of this approach implies that a large amount of data had to be analyzed since it was not possible, in principle, to select only a few parameters among all the potential effects. Several biochemical parameters have been evaluated by comparing their values during the periods of exposure (field ON) and non exposure (field OFF). The evidence that long term exposure to electromagnetic fields with a well defined frequency may have relevant effects on parameters such as body weight, blood glucose and fatty acid metabolism has been obtained. PMID- 18380894 TI - Severe thrombocytosis and anemia associated with celiac disease in a young female patient: a case report. AB - INTRODUCTION: Platelet counts exceeding 1.000 x 103/microl are usually considered secondary to another cause, particularly to chronic myeloproliferative disease (CMPD). Reactive thrombocytosis due to iron deficiency rarely exceeds platelet counts of 700 x 103/microl. CASE PRESENTATION: Here we report the case of a young woman presenting with clinical signs of severe anemia. Laboratory findings confirmed an iron-deficiency anemia associated with severe thrombocytosis of 1703 x 103/microl. Macroscopic gastrointestinal and genitourinary tract bleeding was excluded. The excessive elevation of platelets, slightly elevated lactate dehydrogenase and slightly elevated leukocytes along with the absence of other inflammation parameters raised the suspicion of an underlying hematological disease. However, bone marrow evaluation could not prove the suspected diagnosis of a CMPD, especially essential thrombocythemia (ET). In the further clinical course the platelet count returned to normal after raising the hemoglobin to a level close to normal range with erythrocyte transfusion, and normalization of serum iron and decline of erythropoietin. Finally, following small bowel biopsy, despite the absence of typical clinical signs, celiac disease was diagnosed. After discharge from hospital the patient was commenced on a gluten-free diet and her hemoglobin almost completely normalized in the further follow-up period. CONCLUSION: This case illustrates the rare constellation of an extreme thrombocytosis most likely secondary to iron deficiency due to celiac disease. This represents, to the best of the authors' knowledge, the highest reported platelet count coincident with iron deficiency. A potential mechanism for the association of iron-deficiency anemia and thrombocytosis is discussed. Even in the presence of 'atypically' high platelets one should consider the possibility of reactive thrombocytosis. Extreme thrombocytosis could emerge in the case of iron deficiency secondary to celiac disease. PMID- 18380893 TI - Clinical response after two cycles compared to HER2, Ki-67, p53, and bcl-2 in independently predicting a pathological complete response after preoperative chemotherapy in patients with operable carcinoma of the breast. AB - INTRODUCTION: To investigate the predictive value of clinical and biological markers for a pathological complete remission after a preoperative dose-dense regimen of doxorubicin and docetaxel, with or without tamoxifen, in primary operable breast cancer. METHODS: Patients with a histologically confirmed diagnosis of previously untreated, operable, and measurable primary breast cancer (tumour (T), nodes (N) and metastases (M) score: T2-3(> or = 3 cm) N0-2 M0) were treated in a prospectively randomised trial with four cycles of dose-dense (bi weekly) doxorubicin and docetaxel (ddAT) chemotherapy, with or without tamoxifen, prior to surgery. Clinical and pathological parameters (menopausal status, clinical tumour size and nodal status, grade, and clinical response after two cycles) and a panel of biomarkers (oestrogen and progesterone receptors, Ki-67, human epidermal growth factor receptor 2 (HER2), p53, bcl-2, all detected by immunohistochemistry) were correlated with the detection of a pathological complete response (pCR). RESULTS: A pCR was observed in 9.7% in 248 patients randomised in the study and in 8.6% in the subset of 196 patients with available tumour tissue. Clinically negative axillary lymph nodes, poor tumour differentiation, negative oestrogen receptor status, negative progesterone receptor status, and loss of bcl-2 were significantly predictive for a pCR in a univariate logistic regression model, whereas in a multivariate analysis only the clinical nodal status and hormonal receptor status provided significantly independent information. Backward stepwise logistic regression revealed a response after two cycles, with hormone receptor status and lymph-node status as significant predictors. Patients with a low percentage of cells stained positive for Ki-67 showed a better response when treated with tamoxifen, whereas patients with a high percentage of Ki-67 positive cells did not have an additional benefit when treated with tamoxifen. Tumours overexpressing HER2 showed a similar response to that in HER2-negative patients when treated without tamoxifen, but when HER2-positive tumours were treated with tamoxifen, no pCR was observed. CONCLUSION: Reliable prediction of a pathological complete response after preoperative chemotherapy is not possible with clinical and biological factors routinely determined before start of treatment. The response after two cycles of chemotherapy is a strong but dependent predictor. The only independent factor in this subset of patients was bcl-2. PMID- 18380895 TI - On detection and assessment of statistical significance of Genomic Islands. AB - BACKGROUND: Many of the available methods for detecting Genomic Islands (GIs) in prokaryotic genomes use markers such as transposons, proximal tRNAs, flanking repeats etc., or they use other supervised techniques requiring training datasets. Most of these methods are primarily based on the biases in GC content or codon and amino acid usage of the islands. However, these methods either do not use any formal statistical test of significance or use statistical tests for which the critical values and the P-values are not adequately justified. We propose a method, which is unsupervised in nature and uses Monte-Carlo statistical tests based on randomly selected segments of a chromosome. Such tests are supported by precise statistical distribution theory, and consequently, the resulting P-values are quite reliable for making the decision. RESULTS: Our algorithm (named Design-Island, an acronym for Detection of Statistically Significant Genomic Island) runs in two phases. Some 'putative GIs' are identified in the first phase, and those are refined into smaller segments containing horizontally acquired genes in the refinement phase. This method is applied to Salmonella typhi CT18 genome leading to the discovery of several new pathogenicity, antibiotic resistance and metabolic islands that were missed by earlier methods. Many of these islands contain mobile genetic elements like phage mediated genes, transposons, integrase and IS elements confirming their horizontal acquirement. CONCLUSION: The proposed method is based on statistical tests supported by precise distribution theory and reliable P-values along with a technique for visualizing statistically significant islands. The performance of our method is better than many other well known methods in terms of their sensitivity and accuracy, and in terms of specificity, it is comparable to other methods. PMID- 18380896 TI - JCoast - a biologist-centric software tool for data mining and comparison of prokaryotic (meta)genomes. AB - BACKGROUND: Current sequencing technologies give access to sequence information for genomes and metagenomes at a tremendous speed. Subsequent data processing is mainly performed by automatic pipelines provided by the sequencing centers. Although, standardised workflows are desirable and useful in many respects, rational data mining, comparative genomics, and especially the interpretation of the sequence information in the biological context, demands for intuitive, flexible, and extendable solutions. RESULTS: The JCoast software tool was primarily designed to analyse and compare (meta)genome sequences of prokaryotes. Based on a pre-computed GenDB database project, JCoast offers a flexible graphical user interface (GUI), as well as an application programming interface (API) that facilitates back-end data access. JCoast offers individual, cross genome-, and metagenome analysis, and assists the biologist in exploration of large and complex datasets. CONCLUSION: JCoast combines all functions required for the mining, annotation, and interpretation of (meta)genomic data. The lightweight software solution allows the user to easily take advantage of advanced back-end database structures by providing a programming and graphical user interface to answer biological questions. JCoast is available at the project homepage. PMID- 18380898 TI - Exogenous tumour necrosis factor alpha induces suppression of autoimmune arthritis. AB - INTRODUCTION: Our previous studies showed that arthritic Lewis (LEW) rats produced the highest levels of tumour necrosis factor (TNF)alpha in the recovery phase of adjuvant arthritis (AA), suggesting a correlation between high TNFalpha levels and reduced severity of arthritis. To further explore this correlation, we compared the TNFalpha secretion profile of the AA-resistant Wistar Kyoto (WKY) rats with that of LEW rats, determined the effect of exogenous TNFalpha on the course of AA in LEW rats, and examined various mechanisms involved in TNFalpha induced disease modulation. METHODS: A cohort each of LEW and WKY rats was immunised subcutaneously with heat-killed Mycobacterium tuberculosis H37Ra (Mtb). At different time points thereafter, subgroups of rats were killed and their draining lymph node cells were tested for cytokine production. Another group of LEW rats was injected with TNFalpha intraperitoneally daily for a total of 10 injections, 3 before and 6 after Mtb challenge, and then observed for signs of AA. In parallel, TNFalpha-treated rats were examined for changes in other cytokines, in CD4+CD25+ T cell frequency, and in indoleamine 2,3-dioxygenase (IDO) mRNA expression levels. RESULTS: LEW rats displayed a TNFalpha secretion profile that was opposite to that of the WKY rats. Furthermore, TNFalpha treatment significantly down modulated the severity of AA in LEW rats, and decreased the interferon (IFN)-gamma secretion in response to the pathogenic determinant of the disease-related antigen. No significant alterations were observed in other parameters tested. CONCLUSION: The role of endogenous TNFalpha in the induction and propagation of arthritis is well established. However, exogenous TNFalpha can down modulate the course of AA, displaying an immunoregulatory functional attribute of this cytokine. PMID- 18380897 TI - Evolutionary primacy of sodium bioenergetics. AB - BACKGROUND: The F- and V-type ATPases are rotary molecular machines that couple translocation of protons or sodium ions across the membrane to the synthesis or hydrolysis of ATP. Both the F-type (found in most bacteria and eukaryotic mitochondria and chloroplasts) and V-type (found in archaea, some bacteria, and eukaryotic vacuoles) ATPases can translocate either protons or sodium ions. The prevalent proton-dependent ATPases are generally viewed as the primary form of the enzyme whereas the sodium-translocating ATPases of some prokaryotes are usually construed as an exotic adaptation to survival in extreme environments. RESULTS: We combine structural and phylogenetic analyses to clarify the evolutionary relation between the proton- and sodium-translocating ATPases. A comparison of the structures of the membrane-embedded oligomeric proteolipid rings of sodium-dependent F- and V-ATPases reveals nearly identical sets of amino acids involved in sodium binding. We show that the sodium-dependent ATPases are scattered among proton-dependent ATPases in both the F- and the V-branches of the phylogenetic tree. CONCLUSION: Barring convergent emergence of the same set of ligands in several lineages, these findings indicate that the use of sodium gradient for ATP synthesis is the ancestral modality of membrane bioenergetics. Thus, a primitive, sodium-impermeable but proton-permeable cell membrane that harboured a set of sodium-transporting enzymes appears to have been the evolutionary predecessor of the more structurally demanding proton-tight membranes. The use of proton as the coupling ion appears to be a later innovation that emerged on several independent occasions. PMID- 18380899 TI - Dominant negative Bmp5 mutation reveals key role of BMPs in skeletal response to mechanical stimulation. AB - BACKGROUND: Over a hundred years ago, Wolff originally observed that bone growth and remodeling are exquisitely sensitive to mechanical forces acting on the skeleton. Clinical studies have noted that the size and the strength of bone increase with weight bearing and muscular activity and decrease with bed rest and disuse. Although the processes of mechanotransduction and functional response of bone to mechanical strain have been extensively studied, the molecular signaling mechanisms that mediate the response of bone cells to mechanical stimulation remain unclear. RESULTS: Here, we identify a novel germline mutation at the mouse Bone morphogenetic protein 5 (Bmp5) locus. Genetic analysis shows that the mutation occurs at a site encoding the proteolytic processing sequence of the BMP5 protein and blocks proper processing of BMP5. Anatomic studies reveal that this mutation affects the formation of multiple skeletal features including several muscle-induced skeletal sites in vivo. Biomechanical studies of osteoblasts from these anatomic sites show that the mutation inhibits the proper response of bone cells to mechanical stimulation. CONCLUSION: The results from these genetic, biochemical, and biomechanical studies suggest that BMPs are required not only for skeletal patterning during embryonic development, but also for bone response and remodeling to mechanical stimulation at specific anatomic sites in the skeleton. PMID- 18380900 TI - High incidence of antimicrobial resistant organisms including extended spectrum beta-lactamase producing Enterobacteriaceae and methicillin-resistant Staphylococcus aureus in nasopharyngeal and blood isolates of HIV-infected children from Cape Town, South Africa. AB - BACKGROUND: There is little information on nasopharyngeal (NP) flora or bacteremia in HIV-infected children. Our aim was to describe the organisms and antimicrobial resistance patterns in children enrolled in a prospective study comparing daily and three times weekly trimethoprim-sulfamethoxazole (TMP-SMX) and isoniazid (INH) or placebo prophylaxis. METHODS: NP swabs were taken at baseline from HIV-infected children enrolled in the study. Standard microbiological techniques were used. Children were grouped according to previous or current exposure to TMP-SMX and whether enrolled to the study during a period of hospitalization. Blood culture results were also recorded within 12 months of baseline. RESULTS: Two hundred and three children, median age 1.8 (Interquartile [IQ]: 0.7-4) years had NP swabs submitted for culture. One hundred and eighty four (90.7%) had either stage B or C HIV disease. One hundred and forty-one (69.8%) were receiving TMP-SMX and 19 (9.4%) were on antiretroviral therapy. The majority, 168 (82%) had a history of hospitalization and 91 (44.8%) were enrolled during a period of hospitalization. Thirty-two subjects (16.2%) died within 12 months of study entry. One hundred and eighty-one potential pathogens were found in 167 children. The most commonly isolated organisms were Streptococcus pneumoniae (48: 22.2%), Gram-negative respiratory organisms (Haemophilus influenzae and Moraxella catarrhalis) (47: 21.8%), Staphylococcus aureus (44: 20.4%), Enterobacteriaceae 32 (14.8%) and Pseudomonas 5 (2.3%). Resistance to TMP SMX occurred in > 80% of pathogens except for M. catarrhalis (2: 18.2% of tested organisms). TMP-SMX resistance tended to be higher in those receiving it at baseline (p = 0.065). Carriage of Methicillin resistant S. aureus (MRSA) was significantly associated with being on TMP-SMX at baseline (p = 0.002). Minimal inhibitory concentrations (MIC) to penicillin were determined for 18 S. pneumoniae isolates: 7 (38.9%) were fully sensitive (MIC or=2 microg/ml). Fifty percent of Enterobacteriaceae produced extended spectrum beta-lactamases (ESBL) (resistant to third generation cephalosporins) and 56% were resistant to gentamicin. Seventy-seven percent of S. aureus were MRSA. Carriage of resistant organisms was not associated with hospitalization.On multivariate logistic regression, risk factors for colonization with Enterobacteriaceae were age or= 45 years were identified from PHARMO RLS, including drug-dispensing and hospitalisation data of > 2 million residents of the Netherlands. One-year compliance with bisphosphonates was measured using the Medication Possession Ratio (MPR). To identify determinants of non-compliance, non-compliant women (MPR < 50%) were compared to compliant women (MPR >or= 80%). The effect of patient age, prescriber, initial dosing regimen, gastrointestinal adverse events, co-medication and fractures on non-compliance was investigated. RESULTS: The study cohort included 8822 new users of bisphosphonates, of whom 5079 (58%) were compliant and 2720 (31%) were non-compliant after 1 year. Only 1023 women (11%) had a MPR between >or= 50% and < 80%. Daily dosing at start, increased number of co-medications and new use of intestinal agents in the year after starting bisphosphonates were independently associated with an increased odds of non-compliance. In contrast, higher age, first prescription from a specialist, osteoporosis related hospitalisation and use of NSAIDs in the year preceding bisphosphonate therapy decreased the odds of non-compliance. CONCLUSION: This study revealed several determinants of non-compliance with bisphosphonates, the best controllable being the type of initial bisphosphonate, with daily dosing leading to more non-compliance than weekly dosing. However, compliance for both regimens is suboptimal, pointing to an unmet medical need. PMID- 18380911 TI - Gene therapy for osteosarcoma: steps towards clinical studies. AB - Gene therapy, an applied form of biotechnology, relies on the delivery of foreign DNA into cells. More than 50% of all reported clinical trials for gene therapy are for cancer, though only a scant number for osteosarcoma. Osteosarcoma is a neoplasm afflicting young adults, who in their prime years of life suffer debilitation if not death. The disease is not entirely curable, even with surgery combined with aggressive chemotherapy. Thus, other forms of therapies are being evaluated, including gene therapy. There exist two major forms of gene transfer: viral and non-viral. This review only covers proof-of-principle work carried out in cancer beyond the cell culture stage, in animals. Drawing from the experiences of gene therapy against other cancers, studies for which have already reached the clinical phase, the review discusses potential pitfalls and solutions to enhance gene therapy for osteosarcoma. PMID- 18380912 TI - Enhancement of nortriptyline penetration through human epidermis: influence of chemical enhancers and iontophoresis. AB - Different known percutaneous chemical enhancers and iontophoresis have been tested in-vitro to study their ability to increase transdermal absorption of nortriptyline hydrochloride (20 mg mL(-1)). The chemicals 1-dodecanol, Span 20, Azone, (R)-(+)-limonene or isopropyl myristate were used as an overnight pretreatment at 5% (w/w) in ethanol. Furthermore, isopropyl myristate (20%, w/w) and propylene glycol (15%, w/w) were tested in the same vehicle. Iontophoresis was applied directly to the nortriptyline hydrochloride donor solution for three different concentrations (20, 2 and 0.5 mgmL(-1)). The chemical enhancers slightly increased the nortriptyline transdermal flux but iontophoresis was more efficient. In this case, nortriptyline transdermal flux was concentration dependent, having a higher flux when the concentration was lowered. Therefore, iontophoresis was the most suitable technique to increase transdermal absorption of nortriptyline and it could be an alternative method to provide therapeutic concentrations of this drug in smoking cessation treatment. PMID- 18380913 TI - Effect of chitosan crosslinking on bitterness of artemether using response surface methodology. AB - This work examines the influence of various process parameters on artemether entrapped in crosslinked chitosan microparticles for masking bitterness. A central composite design was used to optimize the experimental conditions for bitterness masking. Critical parameters such as the amounts of artemether, chitosan and crosslinking agent have been studied to evaluate how they affect responses such as incorporation efficiency, particle size and drug release at pH 6.8. The desirability function approach has been used to find the best compromise between the experimental results. The optimized microparticles were characterized by Fourier transform infrared spectroscopy and differential scanning calorimetry. Bitterness score was evaluated by human gustatory sensation test. Multiple linear regression analysis revealed that the crosslinking of chitosan significantly affects incorporation efficiency, particle size and drug release at pH 6.8. The bitterness score of microparticles was decreased to 0, compared with 3+ for pure artemether. The proposed method completed masked the bitter taste of artemether. PMID- 18380914 TI - Effect of ion-pairing and enhancers on scutellarin skin permeability. AB - The aim of this work was to investigate the effect of enhancers and organic amines on the in-vitro percutaneous absorption of the major pharmacologically active compound, scutellarin, obtained from breviscapine extract. The donor vehicle consisted of isopropyl myristate-ethanol in a ratio of 4:1. Percutaneous absorption across full thickness rat skin was investigated in-vitro using 2 chamber diffusion cells, with reverse-phase HPLC for quantification of the permeating scutellarin. Organic amines increased scutellarin permeation by ion pair formation. We also found that the cumulative amount of scutellarin over a period of 12 h of scutellarin was inversely related to the molecular weight of organic amines (r = 0.9134), as well as the logarithm of scutellarin permeability coefficient inversely related to the partition coefficient of organic amines (r = 0.8929). All the permeation enhancers tested increased the cumulative amount of scutellarin over a period of 12 h, and the order of this increase was n-methyl-2 pyrrolidone, oleic acid, menthol or Azone. Drug solubility in donor phase was markedly increased by Azone and n-methyl-2-pyrrolidone, and reduced by menthol and oleic acid. The combined effects of ethanolamine plus Azone, ethanolamine plus menthol, and Azone plus menthol were also investigated. Azone plus menthol had a synergistic effect on the cumulative amount of scutellarin over a period of 12 h. PMID- 18380915 TI - Beneficial effect of antibiotics on ciliary beat frequency of human nasal epithelial cells exposed to bacterial toxins. AB - In the present study, we explored whether the cilio-inhibitory effect induced by toxins derived from bacterial infections could be compensated for by a cilio stimulatory effect of antibiotics. Human nasal epithelial cells (HNEC) expressing beating cilia were grown as monolayers. Ciliary beat frequency (CBF) was determined using an inverted microscope coupled with a high-speed digital camera. Clarithromycin and neomycin did not influence ciliary activity. Bacitracin, clindamycin, gramicidin and roxithromycin increased CBF significantly: by 50 +/- 12%, 54 +/- 16%, 31 +/- 16% and 31 +/- 18%, respectively. A 30 min exposure to Staphylococcus aureus enterotoxin B (SEB) and Pseudomonas aeruginosa lipopolysaccharide (PAL) decreased CBF significantly, by 37 +/- 16 and 28 +/- 12%, respectively. In contrast with exposure to the toxin alone, co-incubation of the nasal monolayer cells with PAL and bacitracin or clindamycin did not result in a decrease in CBF after 30 and 60 min. The effect of SEB could be compensated for by bacitracin but not by clindamycin. After a 12 h preincubation period with SEB, co-incubation with either bacitracin or clindamycin resulted in the complete recovery of CBF. This study suggests that topical antibiotic treatment of nasal infections could result in a dual positive effect, namely treatment of the bacterial infection and recovery of ciliary activity. PMID- 18380916 TI - Role of maltol in advanced glycation end products and free radicals: in-vitro and in-vivo studies. AB - Inhibitors of advanced glycation end products (AGEs) have potential as preventive agents against diabetic complications. In-vitro AGE inhibitory activity, transition metal chelating, and free radical scavenging activity tests have been used to screen for and identify effective AGE inhibitors. In an ongoing project to elucidate AGE inhibiting active components of heat-processed ginseng, maltol was selected for more detailed investigation. Although there are several lines of evidence concerning the antioxidant activity of maltol, the in-vitro and in-vivo inhibitory effects of maltol on AGE generation have not been evaluated. In the present study, the in-vitro AGE inhibitory effects and free radical scavenging activity of maltol were investigated. In addition, the in-vivo therapeutic potential of maltol against diabetic renal damage was tested using streptozotocin (STZ)-diabetic rats. Maltol showed a stronger AGE inhibitory effect than aminoguanidine, a well known AGE inhibitor. In addition, the hydroxyl radical scavenging activity of maltol on electron spin resonance (ESR) spectrometry was slightly stronger than that of aminoguanidine. Therefore, maltol was found to have stronger in-vitro AGE inhibiting activity compared with aminoguanidine. The administration of 50 mgkg(-1) per day of maltol suppressed the elevated serum levels of glycosylated protein, renal fluorescent AGEs, carboxymethyllysine, receptors for AGEs, and nuclear factor-kappaB p65 in diabetic control rats. These beneficial effects of maltol against STZ-diabetic renal damage were thought to result from its free radical scavenging and AGE inhibitory effects. PMID- 18380917 TI - Pimaradienoic acid inhibits vascular contraction and induces hypotension in normotensive rats. AB - The present investigation was designed to investigate the effect of the diterpene ent-pimara-8(14),15-dien-19-oic acid (pimaradienoic acid, PA) on smooth muscle extracellular Ca(2+) influx. To this end, the effect of PA on phenylephrine- and KCl-induced increases in cytosolic calcium concentration ([Ca(2+)](c)), measured by the variation in the ratio of fluorescence intensities (R340/380 nm) of Fura 2, was analysed. Whether bolus injection of PA could induce hypotensive responses in conscious normotensive rats was also evaluated. PA inhibited the contraction induced by phenylephrine (0.03 or 10 micromol L(-1)) and KCl (30 or 90 mmol L( 1)) in endothelium-denuded rat aortic rings in a concentration dependent manner. Pre-treatment with PA (10, 100, 200 micromol L(-1)) attenuated the contraction induced by CaCl(2) (0.5 nmol L(-1) or 2.5 mmol L(-1)) in denuded rat aorta exposed to Ca(2+)-free medium containing phenylephrine (0.1 micro mol L(-1)) or KCl (30 mmol L(-1)). Interestingly, the inhibitory effect displayed by PA on CaCl(2)-induced contraction was more pronounced when KCl was used as the stimulant. Phenylephrine- and KCl-induced increases in [Ca(2+)](c) were inhibited by PA. Similarly, verapamil, a Ca(2+)-channel blocker, also inhibited the increase in [Ca(2+)](c) induced by either phenylephrine or KCl. Finally, bolus injection of PA (1-15 mg kg(-1)) produced a dose-dependent decrease in mean arterial pressure in conscious normotensive rats. The results provide the first direct evidence that PA reduces vascular contractility by reducing extracellular Ca(2+) influx through smooth muscle cellular membrane, a mechanism that could mediate the hypotensive response induced by this diterpene in normotensive rats. PMID- 18380918 TI - Soybean fat supplementation controls insulin resistance caused by fat-free total parenteral nutrition. AB - Conscious young adult male rats were given total parenteral nutrition (TPN) with or without soybean fat for 4 days. Those given fat-free TPN developed severe fatty liver, with hyperglycaemia, hyperinsulinaemia, and hypotriglyceridaemia. These disorders were clearly improved by supplementing TPN with soybean fat, in an amount equivalent to 20% of total calories, and correspondingly reducing glucose. Insulin resistance also developed over a 4-day infusion of fat-free TPN in mature rats. Even after over-night fasting after stopping the TPN infusion, the levels of serum glucose and insulin were higher in the fat-free TPN group than in the control group, and intravenous glucose tolerance test results indicated insulin resistance in the fat-free TPN group. The HOMA-IR index of insulin resistance was significantly improved by supplementation with soybean fat. In conclusion, fat-free TPN infusion induced hyperglycaemia and hyperinsulinaemia, leading to fatty liver and insulin resistance. TPN with glucose should be supplemented with soybean fat emulsion as replacement for part of the glucose calories. PMID- 18380919 TI - Pharmacological effect of capsaicin on rat avoidance behaviours elicited by sine wave electrical stimulation of different frequencies by Neurometer. AB - The Neurometer is a diagnostic device for measuring the perception and threshold of transcutaneous stimulation. It has been used in patients to selectively activate Abeta-, Adelta- and C-fibres in the primary afferents at different stimulus frequencies (2000, 250 and 5 Hz, respectively). In this study, we investigated use of the Neurometer to selectively activate nerves in conscious rats. The behavioural endpoint of paw withdrawal was used to measure the current threshold (CT). This behaviour was elicited by a lower stimulus current than other behaviours evoked by Neurometer stimulation and caused only mild stress in rats. Repeated topical application of capsaicin (four doses of 100 microg) or systemic administration of the capsaicin analogue resiniferatoxin (30 or 300 microg kg(-1)) increased the CT value for this behaviour at 5 Hz stimulation but not at 2000 Hz or 250 Hz. This change in CT at 5 Hz is probably due to C-fibre desensitization by the pharmacological treatments. The combination of 5 Hz sine wave stimulation with a Neurometer and the observation of paw withdrawal behaviour make it possible to perform preclinical studies of C-fibres in animals as an alternative to the use of high- and low-rate heating of the paw. PMID- 18380920 TI - Anti-inflammatory effects of red pepper (Capsicum baccatum) on carrageenan- and antigen-induced inflammation. AB - Inflammation is a pivotal component of a variety of diseases, such as atherosclerosis and tumour progression. Various naturally occurring phytochemicals exhibit anti-inflammatory activity and are considered to be potential drug candidates against inflammation-related pathological processes. Capsicum baccatum L. var. pendulum (Willd.) Eshbaugh (Solanaceae) is the most consumed species in Brazil, and its compounds, such as capsaicinoids, have been found to inhibit the inflammatory process. However, the anti-inflammatory effects of C. baccatum have not been characterized. Thus, this study was designed to evaluate the effects of C. baccatum juice in animal models of acute inflammation induced by carrageenan and immune inflammation induced by methylated bovine serum albumin. Pretreatment (30 min) of rats with pepper juice (0.25-2.0 g kg(-1)) significantly decreased leucocyte and neutrophil migration, exudate volume and protein and LDH concentration in pleural exudates of a pleurisy model. This juice also inhibited neutrophil migration and reduced the vascular permeability on carrageenan-induced peritonitis in mice. C. baccatum juice also reduced neutrophil recruitment and exudate levels of pro-inflammatory cytokines TNF-alpha and IL-1beta in mouse inflammatory immune peritonitis. Furthermore, we demonstrated that the main constituent of C. baccatum juice, as extracted with chloroform, is capsaicin. In agreement with this, capsaicin was able to inhibit the neutrophil migration towards the inflammatory focus. To our knowledge, this is the first demonstration of the anti-inflammatory effect of C. baccatum juice and our data suggest that this effect may be induced by capsaicin. Moreover, the anti-inflammatory effect induced by red pepper may be by inhibition of pro inflammatory cytokine production at the inflammatory site. PMID- 18380921 TI - Topical anti-inflammatory activity of Eugenia brasiliensis Lam. (Myrtaceae) leaves. AB - Eugenia brasiliensis Lam., a plant from the south of Brazil, is used in the popular medicine for rheumatism treatment. This study reports that topical application of hydroalcoholic extract, fractions and isolated compounds from E. brasiliensis caused an inhibition of ear oedema in response to topical application of croton oil on the mouse ear. For oedema inhibition, the estimated ID50 values (dose reducing the inflammatory response by 50% relative to the control value) for hydroalcoholic extract and fractions (hexane, ethyl acetate and dichloromethane) were 0.17, 0.29, 0.13 and 0.14 mg/ear, respectively, with inhibition of 79+/-7%, 87+/-6%, 88+/-5% and 96+/-2%, respectively. Isolated phenolic compounds (quercetin, catechin and gallocatechin) were also effective in inhibiting the oedema (inhibition of 61+/-5%, 66+/-2% and 37+/-9%, respectively). Moreover, both extract and isolated compounds caused inhibition of polymorphonuclear cells influx (inhibition of 85+/-6%, 81+/-5%, 73+/-6% and 76+/ 6%, respectively). The histological analysis of the ear tissue clearly confirmed that the extract and compounds of E. brasiliensis inhibited the influx of polymorphonuclear cells to mouse ear skin after application of croton oil. Furthermore, hydroalcoholic extract was also effective in inhibiting the arachidonic acid-mediated mouse ear oedema (ID50 value was 1.94 mg/ear and inhibition of 60+/-7%). Therefore, these results consistently support the notion that E. brasiliensis possesses topical anti-inflammatory activity. PMID- 18380922 TI - Vascular myorelaxing activity of isolates from South African Hyacinthaceae partly mediated by activation of soluble guanylyl cyclase in rat aortic ring preparations. AB - The vasorelaxing effect of isolates (compounds 1, 2, 3, and 4 (homoisoflavanones), compound 5 (sesquiterpenoid), compounds 6 and 7 (bufadienolides)) from the South African Hyacinthaceae has been assessed using rat aortic ring preparations. Compounds 2, 3, and 4 inhibited the tonic contraction induced by both 60 mM K(+) (K60) and phenylephrine, compound 3 being the most potent. Compounds 5, 6, and 7 caused a modest concentration-dependent relaxation, whereas compound 1 was ineffective. Under K25- or K60-induced depolarization, compound 3 displayed antispasmodic effects not reversed by tetraethylammonium. Under precontraction induced with phenylephrine, compound 3 shifted to the left the concentration-relaxation curves of either isoprenaline or sodium nitroprusside. 1 H-[1,2,4] oxidazolol [4,3-a] quinoxalin-1-one shifted to the right the concentration-relaxation curve of compound 3, while 3'-isobutyl-1 methylxanthine had no effect. In the absence of extracellular Ca(2+), compound 3 (estimated pIC50 = 4.66) and ryanodine reduced the response to phenylephrine. Phenylephrine-stimulated influx of extracellular Ca(2+) was markedly reduced when tissues were pretreated with compound 3 (pIC50 = 5.14) or nifedipine, but stimulated by ryanodine. Compound 3 partially antagonized the contraction induced by phorbol 12-myristate-13-acetate. To our knowledge, this has been the first account describing the vasodilating activity of homoisoflavonoids: compound 3 proved an effective vasorelaxing agent, partly acting via the activation of soluble guanylyl cyclase. PMID- 18380923 TI - Induction of hepatocyte growth factor by fucoidan and fucoidan-derived oligosaccharides. AB - Fucoidan, which is extracted from brown seaweed, is a complex sulphated polysaccharide that is mostly composed of L-fucose and sulphated ester groups. The structural and anionic characteristics of fucoidan are similar to those of heparin. Heparin stimulates production of hepatocyte growth factor (HGF), which has key roles in tissue regeneration. We have shown that fucoidan and fucoidan derived oligosaccharides have similar ability to stimulate production of HGF as heparin and heparin-derived oligosaccharides. This induction of HGF by heparin or fucoidan and their oligosaccharide derivates occurs primarily at the level of translation, probably via the same mechanism. Fucoidan may thus be useful to protect tissues and organs from various injuries and diseases, via mechanisms involving HGF. PMID- 18380924 TI - Effect of methanolic extract of Terminalia arjuna against Helicobacter pylori 26695 lipopolysaccharide-induced gastric ulcer in rats. AB - Helicobacter pylori lipopolysaccharide (HP-LPS) is a potent virulence factor in the causation of gastric ulcer and gastritis. H. pylori-induced gastric pathology is prevalent throughout the world. Herbal medicines are attracting attention because of their traditional values, popularity and belief, as well as for their advantages such as less toxicity, affordability and medicinal value. The present study aimed to evaluate the anti-ulcer effect of a methanolic extract of Terminalia arjuna (TA) against HP-LPS-induced gastric damage in rats. Ulcers were induced with HP-LPS (50 mug per animal) administered orally daily for 3 days. The efficacy of TA on gastric secretory parameters such as volume of gastric juice, pH, free and total acidity, pepsin concentration, and the cytoprotective parameters such as protein-bound carbohydrate complexes in gastric juice and gastric mucosa was assessed. The protective effect of TA was also confirmed by histopathological examination of gastric mucosa. HP-LPS-induced alterations in gastric secretory parameters were altered favourably in rats treated with TA, suggesting that TA has an anti-secretory role. Furthermore, HP-LPS-induced impairments in gastric defence factors were also prevented by treatment with TA. These results suggest that the severe cellular damage and pathological changes caused by HP-LPS are mitigated by TA; these effects are comparable with those of sucralfate. The anti-ulcer effect of TA may reflect its ability to combat factors that damage the gastric mucosa, and to protect the mucosal defensive factors. PMID- 18380925 TI - The anti-inflammatory modulatory role of Solidago chilensis Meyen in the murine model of the air pouch. AB - The aim of this study was to investigate the anti-inflammatory efficacy of an aqueous extract (AE), and its butanolic (BuOH) and aqueous residual (AR) fractions, derived from the rhizome of Solidago chilensis in inflammation caused by carrageenan in mice. Solidago chilensis Meyen rhizome was extracted using hot water at 90 degrees C under infusion. The extract was filtered and lyophilized. Part of the aqueous extract was fractionated with n-BuOH, resulting in butanolic (BuOH) and aqueous residual (AR) fractions. Adult Swiss mice were used in the in vivo experiments. We evaluated the effect of rhizome aqueous extract of Solidago chilensis and these two derived fractions on the inflammation induced by carrageenan in the mouse model of the air pouch. The aqueous extract and its derived fractions significantly inhibited leucocytes, neutrophils, exudation, myeloperoxidase and adenosine deaminase activity, as well as nitric oxide, interleukin-1 beta (IL-1beta), neutrophil chemokine (KC) and tumour necrosis factor-alpha (TNF-alpha) levels (P < 0.05). Indometacin and dexamethasone inhibited all the studied inflammatory parameters (P < 0.01) with the exceptions that indometacin did not inhibit TNF-alpha levels and dexamethasone did not inhibit KC levels (P > 0.05). These results indicate that Solidago chilensis has a significant anti-inflammatory action on acute inflammatory responses and that its inhibitory activity may be due not only to the inhibition of proinflammatory mediators, but also to the inhibition of leucocyte infiltration. PMID- 18380926 TI - Methanol extract of Desmodium gangeticum roots preserves mitochondrial respiratory enzymes, protecting rat heart against oxidative stress induced by reperfusion injury. AB - Ischaemia and reperfusion result in mitochondrial dysfunction, with decreased oxidative capacity, loss of cytochrome c and generation of reactive oxygen species. The aim of this study was to evaluate the effect of a methanol extract of Desmodium gangeticum (L) DC (Fabaceae) (DG) on lipid peroxidation and antioxidants in mitochondria and tissue homogenates of normal, ischaemic and ischaemia-reperfused rats. Myocardial lipid peroxidation products (thiobarbituric acid reactive substances; TBARS) in cardiac tissue homogenates and mitochondrial fractions were significantly increased during ischaemia reperfusion. Antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase (GPx) and glutathione reductase) in the myocardial tissue homogenate and mitochondria decreased significantly during ischaemia reperfusion, accompanied by a decreased activity of mitochondrial respiratory enzymes. Daily pretreatment of rats with DG (50 or 100 mgkg(-1)) orally for 30 days had a significant effect on the activity of mitochondrial and antioxidant enzymes. In-vitro studies showed that DG inhibited lipid peroxidation, and also scavenged hydroxyl and superoxide radicals. The concentrations required to scavenge 50% of the superoxide and hydroxyl radicals were 21 and 50.5 microgmL(-1), respectively. Administration of DG to normal rats did not have any significant effect on any of the parameters studied. The results of our study showed that DG possesses the ability to scavenge the free radicals generated during ischaemia and ischaemia reperfusion and thereby preserves the mitochondrial respiratory enzymes that eventually lead to cardioprotection. PMID- 18380927 TI - A list of enzyme preparations covered by the term enzybiotics should not be restricted to bacteriophage-encoded peptidoglycan hydrolases (lysins). PMID- 18380929 TI - Evidence-based therapy of severe acute respiratory distress syndrome: an algorithm-guided approach. AB - Despite considerable research and constantly emerging treatment modalities, the mortality associated with acute respiratory distress syndrome (ARDS) has remained virtually unchanged over the last decade. Clinical studies have been unable to show a reduction in mortality for most therapeutic interventions except for low tidal volume ventilation. Failure to prove a mortality benefit might be a result of the varying severity of ARDS in the patients studied. Nevertheless, positive responses to single supportive measures (inhaled nitric oxide, prone positioning and extracorporeal membrane oxygenation) have been demonstrated in multiple trials. Criteria for administration, weaning and discontinuation of these supportive interventions have never been described in detail. In this context, implementation of an evidence-based algorithm might facilitate clinical management of severe ARDS. This review summarizes the current evidence base and proposes a new treatment algorithm that aims to prioritize the administration of advanced strategies in a multimodal approach for ARDS. PMID- 18380930 TI - MRI screening of dysbaric osteonecrosis in hyperbaric-chamber inside attendants. AB - Inside attendants are medical staff who accompany patients during hyperbaric oxygen treatments. Dysbaric osteonecrosis (DON) is a well-known consequence of hyperbaric exposure. The aim of this study was to evaluate DON in inside attendants using magnetic resonance imaging (MRI). The bilateral shoulder, hip and knee joints of 12 inside attendants (four men, eight women; mean age 29 years; age range 22 - 36 years) were investigated. The mean +/- SD duration of employment as an inside attendant was 3.8 +/- 3.0 years (range 1 - 9 years) and the mean +/- SD number of hyperbaric exposures was 198 +/- 267 (median 96; range 30 - 950). None of the inside attendants had a history of decompression sickness. The MRIs of the attendants did not reveal bone lesions consistent with DON. This study failed to find an increased risk for DON in inside attendants. Additional multicentre epidemiological studies are warranted to investigate the occupational safety of inside attendants. PMID- 18380931 TI - Using 100% oxygen does not alter the cardiovascular autonomic regulation during non-invasively simulated haemorrhage in healthy volunteers. AB - We tested the effect of 100% oxygen on heart rate (HR), arterial blood pressure (ABP), cardiac output (CO), stroke volume (SV), total peripheral resistance (TPR), HR variability (HRV), systolic blood pressure variability (SBPV) and baroreflex sensitivity (BRS) in 20 healthy volunteers during simulated haemorrhage induced by -40 mmHg lower body negative pressure (LBNP). HRV in the high frequency region (HRV HF), BRS, ABP and TPR were significantly increased, SBPV in the low frequency region (SBPV LF), CO and SV were unchanged, and HR was significantly decreased by 100% oxygen administration during normovolaemia. HRV HF, BRS, CO and SV were significantly decreased, SBPV LF and ABP were unchanged, and HR and TPR were significantly increased by LBNP during 21% or 100% oxygen administration. There were no significant differences in cardiovascular autonomic and haemodynamic responses to LBNP during 21% or 100% oxygen administration, suggesting that 100% oxygen does not alter normal cardiovascular autonomic responses during simulated haemorrhage. PMID- 18380932 TI - Telmisartan, an angiotensin II type 1 receptor blocker, inhibits advanced glycation end-product (AGE)-elicited hepatic insulin resistance via peroxisome proliferator-activated receptor-gamma activation. AB - This study examined whether telmisartan, a unique angiotensin II type 1 receptor blocker (ARB) with peroxisome proliferator-activated receptor-gamma (PPAR-gamma) modulating activity, improved insulin resistance in advanced glycation end product (AGE)-exposed human hepatoma (Hep3B) cells. AGE increased phosphorylation of insulin receptor substrate-1 (IRS-1) at serine-307 residues in Hep3B cells. It also decreased tyrosine phosphorylation of IRS-1 and, subsequently, reduced the association of the p85 subunit of phosphatidylinositol 3-kinase with IRS-1 and glycogen synthesis in insulin-exposed Hep3B cells, all of which were inhibited by telmisartan. The insulin-sensitizing properties of telmisartan in AGE-exposed Hep3B cells were significantly blocked by GW9662, an inhibitor of PPAR-gamma. Candesartan, another ARB, did not affect AGEs-induced serine phosphorylation of IRS-1 at serine-307 residues in Hep3B cells. Our study suggests that telmisartan could improve AGE-elicited insulin resistance in Hep3B cells by inhibiting serine phosphorylation of IRS-1, at least in part, via activation of PPAR-gamma. Telmisartan may play a protective role against hepatic insulin resistance in diabetes. PMID- 18380933 TI - Tamsulosin treatment of chronic non-bacterial prostatitis. AB - The efficacy of tamsulosin in the treatment of chronic non-bacterial prostatitis was evaluated in a randomized clinical observation of 105 male outpatients conducted for 90 days. Patients were randomly divided into five groups (n = 21 per group) according to prostatitis type IIIA or IIIB and therapy regimens (tamsulosin, levofloxacin, or tamsulosin plus levofloxacin combination therapy). National Institutes of Health Chronic Prostatitis Symptom Index scores, expressed prostatic massage test and urodynamic urethral pressure and urethral closure pressure tests were performed to evaluate clinical efficacy of the treatments. Scores for pain, urinary symptoms and quality of life were significantly improved by days 45 and 90 after all treatments in both prostatitis categories. Improvements in symptom scores in the combined treatment group were significantly superior to those in the single treatment groups. Tamsulosin and levofloxacin are both effective in the treatment of, and may have an additive effect in, the treatment of non-bacterial prostatitis. PMID- 18380934 TI - Anterior cruciate ligament reconstruction by using otogenous [correction of otogeneous] hamstring tendons with home-based rehabilitation. AB - We investigated patients undergoing arthroscopic anterior cruciate ligament (ACL) reconstruction using otogenous hamstring tendons with a cross-pin technique to compare a post-operative home-based rehabilitation programme with a clinic-based programme. ACL reconstruction was performed on 104 patients (103 male) by the same surgeon. The mean age of the patients was 31.5 years (range 18 - 43 years) and the mean time interval between injury and operation was 34.3 months. Patients were randomly allocated to either a home-based (n = 52) or clinic-based rehabilitation programme (n = 52). Mean follow-up was 31.1 months (range 12 - 66 months). Patients underwent a series of examinations before and after surgery in order to evaluate functional recovery of their injured knee. The results demonstrated that using otogeneous hamstring tendons for ACL reconstruction was safe and produced satisfactory results. The study also demonstrated that a home based rehabilitation programme was as effective as a clinic-based programme. PMID- 18380935 TI - Does the atopic phenotype prevent development of active tuberculosis infection? AB - This study investigated the relationship between the atopic phenotype and the development of active tuberculosis. A total of 82 human immunodeficiency virus negative males with active pulmonary tuberculosis and 88 healthy controls were enrolled into this prospective study. Serum immunoglobulin E (IgE) levels were measured and skin prick tests performed before initiation of treatment. Skin prick tests were positive in 34.1% of the tuberculosis patients and 39.8% of the controls. Allergic respiratory symptoms were significantly less frequent in skin prick test positive tuberculosis patients (21.4%) compared with skin prick test positive controls (62.9%). Median IgE levels in atopic tuberculosis patients were significantly higher than in atopic controls. The low rate of atopic respiratory complaints seen in the tuberculosis patients, despite having similar allergic skin prick test sensitivities to the controls, could be attributed to a weak T helper (Th) 2 immune reaction and its effects on Th1-Th2 interaction. PMID- 18380936 TI - Prevalence of premenstrual syndrome in autism: a prospective observer-rated study. AB - A systematic, prospective observer-rated study was carried out to determine the prevalence of late luteal phase dysphoric disorder (premenstrual syndrome) in women with autism. A group of women with autism and learning disability (n = 26) was compared with a group of women with a non-autism learning disability (n = 36) matched for age, in-patient status, intelligence, marital status, parity, behavioural problems and ethnicity. Observers rated DSM-IV symptoms of late luteal phase dysphoric disorder every day from each subject over three consecutive menstrual cycles. Using a premenstrual increase in DSM-IV symptoms of >or= 30% as evidence of fulfilment of diagnostic criteria, the prevalence of late luteal phase dysphoric disorder was 92% in the autism group compared with 11% in the control group. This difference was highly statistically significant. The principal conclusion from this study is that there is a marked increase in premenstrual syndrome in women with autism compared with matched controls. PMID- 18380937 TI - Increased expression of paxillin is found in human oesophageal squamous cell carcinoma: a tissue microarray study. AB - Oesophageal cancer is one of the most common cancers worldwide. Currently, the tumour, node, metastasis (TNM) staging system is the primary method for determining its extent and prognosis, however, data suggest this system does not predict prognosis accurately. Research has, therefore, concentrated on searching for specific biomarkers. Paxillin has been shown to play an important role in controlling cell spread and migration. Its over-expression is considered to correlate with the prognosis of some types of cancers, however, the relationship between paxillin expression and clinical outcome in oesophageal cancer has not been investigated. This study determined the expression of paxillin by immunohistochemistry on the tissue microarray of 100 oesophageal squamous cell cancer patients followed up for a mean of 55 months. Paxillin was over-expressed in tumours in 27/100 cases, compared with 6/100 cases for adjacent non-tumoural cells. No correlation occurred between expression of paxillin and overall patient survival, hence paxillin is not an effective prognostic marker in these patients. PMID- 18380938 TI - Human gastric adenocarcinoma allelotype on chromosomes 17 and 18. AB - Allelic losses of multiple chromosome loci in gastric adenocarcinoma suggest that inactivation of tumour suppressor genes in these regions may be important for tumourigenesis. To define deletion intervals and find candidate tumour suppressor genes involved in gastric adenocarcinoma pathogenesis, a genome-wide search for loss of heterozygosity (LOH) was conducted in 45 patients with primary gastric adenocarcinoma. Investigations using 29 microsatellite markers spanning chromosomes 17 and 18 showed allelic deletion in 29 (64%) specimens at one or more loci. Five LOH overlap regions, three newly identified as deletion regions, were defined: RI, D17S831 - D17S921 at 17p12-13.3; RII, D17S1868 - D17S787 at 17q21.3-22; RIII, D17S785 - D17S928 at 17q25.3; RIV, D18S61 - D18S1161 at 18q22; and RV, D18S462 - D18S70 at 18q22-q23. Eleven (24%) patients with chromosome 17 allelic loss also showed LOH on 18q, with at least one region of overlapping. LOH mapping showed allelic losses were widespread on both chromosomes and suggests the possibility that multiple tumour suppressor genes, including one or more that are unknown, might be inactivated in the aetiology of gastric adenocarcinoma. PMID- 18380939 TI - The relationship between renal injury and change in vitamin D metabolism in aged rats with insulin resistance or type 2 diabetes mellitus. AB - Insulin resistance (IR), IR treated with vitamin D, IR treated with 1alpha hydroxyvitamin D (1alpha-(OH)D), type 2 diabetes mellitus (T2DM), T2DM treated with vitamin D and T2DM treated with 1alpha-(OH)D were studied in animal models using aged Wistar rats. Glucose infusion rates and levels of urinary albumin (UA), serum 25-hydroxyvitamin D (25-(OH)D) and 1,25-dihydroxyvitamin D (1,25(OH)2D) were measured. T2DM rats had higher UA than IR or normal rats, and levels of 25-(OH)D in all models weresimilar. IR rats had higher 1,25-(OH)2D levels than T2DM rats, and had lower 1,25-(OH)2D levels than normal rats. Treating IR or T2DM rats with vitamin D had no effect on 25-(OH)D or 1,25-(OH)2D. Administration of 1alpha-(OH)D significantly increased 1,25-(OH)2D in IR rats to above-normal levels, and significantly increased 1,25-(OH)2D in T2DM rats to normal levels. In IR or T2DM, abnormal vitamin D metabolism is characterized by 1,25-(OH)2D deficiency and is related to renal injury. PMID- 18380940 TI - Influence of vaccination and surgery on HLA-DR expression in patients with upper aerodigestive tract cancer. AB - Major surgery is associated with an increased risk of post-operative immunosuppression and infections. We investigated the influence of influenza vaccination on cell-mediated immune responses in cancer patients undergoing either surgical or conservative therapy. Forty patients with an upper aerodigestive tract tumour were allocated to either a surgical or non-surgical treatment course. Patients within each group were randomized to the vaccination or non-vaccination group. Vaccination was performed twice before surgery or conservative treatment. Human leucocyte antigen receptor (HLA-DR) expression on monocytes was analysed by flow cytometry. In the surgical patients, HLA-DR expression on day 1 after surgery decreased in both the vaccinated and non vaccinated groups. Vaccinated non-surgical patients showed significantly increased HLA-DR expression levels compared with the non-vaccinated patients. This pilot study demonstrated that vaccination increased monocyte HLA-DR expression in conservatively-treated cancer patients whereas surgery abrogated this response. Vaccination before surgery, therefore, might not help to maintain immune reactivity after surgery. PMID- 18380941 TI - Treatment of intracerebral glioblastomas with G422 tumour cell vaccine in a mouse model. AB - The aim of this study was to develop a tumour vaccine with the ability to induce and expand higher affinity cytotoxic T lymphocytes and stimulate an effective antitumour immune response. The hypothesis tested was that G422 glioblastoma cells modified with B7-1 and interferon (IFN)-gamma genes could serve as a tumour vaccine. It was found that therapeutic subcutaneous immunizations with this tumour vaccine significantly induced a cytotoxic T-cell response and prolonged the survival of female Kuming mice with intracerebral G422 tumour isografts. The data collectively suggested that G422 glioblastoma cells genetically modified with B7-1 and IFN-gamma genes could serve as a tumour vaccine. PMID- 18380942 TI - Anatomical variations of the coronary sinus ostium area of the human heart. AB - Classic anatomical dissection of 150 heart specimens from adults aged 18 - 80 years was performed. The Thebesian valve was absent in 20% of cases and, in these, 4% had a large ostial valve of the middle cardiac vein in front of the coronary sinus ostium. Fibres of Chiari were found in 10% of cases. Ostia of the middle cardiac vein, posterior veins of the left ventricle, small cardiac vein and deep cardiac veins were present in the distal 10 mm of the coronary sinus. Some samples had ostial and/or parietal valves or antivalves that sometimes contained muscular fibres. Distal accessory parietal valves (2%) and antivalves (1%) of the coronary sinus wall were found at a distance of 4 - 7 mm from its ostium. The frequency and variability of anatomical structures in the area of the coronary sinus ostium probably influence the haemodynamics of this area. Knowledge of and being able to identify these anatomical variations may help in identifying and overcoming potential difficulties in treating arrythmias and in cardiosurgery. PMID- 18380943 TI - Gabexate mesilate suppresses influenza pneumonia in mice through inhibition of cytokines. AB - Gabexate mesilate is a synthetic protease inhibitor that is effective for acute pancreatitis. The effect of gabexate mesilate in influenza pneumonia in mice was investigated by examining the changes in pulmonary inflammatory cytokines and chemokines. Pathological changes in the lungs of treated mice were extremely mild, compared with changes in infected, untreated mice. Intrapulmonary levels of interleukin-6 and macrophage inflammatory protein-2 decreased in treated mice compared with untreated mice, despite similar viral titres in the lungs. Survival terms for treated and untreated groups were similar. These data indicate that gabexate mesilate has beneficial effects on influenza pneumonia, which may be due to the modulation of inflammatory cytokine/chemokine responses. PMID- 18380944 TI - Effect of rosiglitazone on cardiac fibroblast proliferation, nitric oxide production and connective tissue growth factor expression induced by advanced glycation end-products. AB - This study investigated the effects of the peroxisome proliferator-activated receptor-gamma agonist rosiglitazone (RGZ) on cardiac fibroblast proliferation, nitric oxide content and connective tissue growth factor (CTGF) expression following incubation with advanced glycation end-products (AGEs). Cultured neonatal rat cardiac fibroblasts were incubated with various concentrations of AGEs for 48 h. Cells were also incubated with 200 mg/l AGEs plus various concentrations of RGZ. Cardiac fibroblast proliferation and cell cycle status were detected using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Western blotting was used to measure the expression of CTGF and nitric oxide content was evaluated using a nitrate reductase assay. AGEs significantly accelerated proliferation, increased CTGF expression and decreased nitric oxide production in cardiac fibroblasts. These effects of AGEs were inhibited by RGZ in a dose-dependent manner. Treatment with RGZ could be a valuable therapeutic approach in diabetic patients with myocardial fibrosis. PMID- 18380945 TI - Effects of telomerase activity on carcinogenesis and survival in laryngeal carcinoma. AB - This study aimed to look at whether a correlation exists between telomerase activity and survival of laryngeal carcinoma patients. Telomerase activity was measured by telomerase repeat amplification protocol in 31 laryngeal carcinomas and adjacent normal tissues, and in 21 vocal cord polyps (controls). Follow-up was for at least 60 months. Telomerase activity in tissues adjacent to laryngeal carcinomas was significantly higher than in the carcinomas which was, in turn, significantly higher than in vocal cord polyps. There was no significant difference between telomerase activity in carcinomas or adjacent tissues and clinicopathological characteristics. Patients with high telomerase activity in carcinoma tissue had significantly shorter survival times than those with low activity, but no significance difference was observed between survival time and telomerase status in adjacent tissues. Linear regression showed significant association between telomerase activity levels in carcinoma tissues and survival time, but this was not observed in adjacent tissues. This study suggests that telomerase activation probably takes place before the cancer phenotype develops and has prognostic significance for survival of patients with laryngeal carcinoma. PMID- 18380946 TI - Conversion from standard opioid therapy to once-daily oral extended-release hydromorphone in patients with chronic cancer pain. AB - This open-label, multicenter study assessed the efficacy and tolerability of conversion to once-daily OROS hydromorphone from previous opioid agonist therapy in patients with chronic cancer pain. Patients were stabilized on their previous therapy before conversion at a 5:1 ratio of morphine sulfate to hydromorphone hydrochloride. The OROS hydromorphone dose was titrated over 3 - 21 days to achieve effective analgesia and was maintained for up to 14 days. Efficacy was assessed using the Brief Pain Inventory (BPI). Adverse events and vital signs were monitored. Dose stabilization was achieved in 119 of the 127 (94%) patients who received the study medication; in 77%, stabilization was achieved with no titration steps. Mean BPI pain intensity ratings and BPI pain interference scores decreased significantly after OROS hydromorphone treatment compared with pretreatment values. Mean pain-relief level remained stable after conversion and throughout treatment with OROS hydromorphone. Adverse events were as expected for cancer patients receiving opioid agonists. There were no clinically significant changes in vital signs. PMID- 18380948 TI - Clinical and genetic analysis of a Korean patient with late-onset X-linked adrenal hypoplasia congenita and hypogonadotropic hypogonadism: identification of a novel mutation in the NR0B1 gene. AB - Adrenal hypoplasia congenita (AHC) is caused by mutations in the NR0B1 gene on chromosome Xp21.3-p21.2. It manifests as X-linked primary adrenal failure in early infancy or childhood and as hypogonadotropic hypogonadism (HHG) at puberty. Although studies of AHC patients and mutations in the NR0B1 gene have been reported throughout the world, there has previously been only one other case report from Korea. We encountered a 23-year old Korean male with delayed-onset AHC/HHG who had been previously diagnosed with adrenal insufficiency of unknown aetiology at age 13 years. Delayed puberty and incomplete HHG were observed. Direct sequencing of the NR0B1 gene revealed the patient to have a novel insertion mutation (c.959_960insT; Leu321ProfsX68). Although AHC is believed to be rare, it should be considered in a differential diagnosis of patients showing late-onset primary adrenal insufficiency. PMID- 18380947 TI - Prevalence of and risk factors for isolated systolic hypertension in the rural adult population of Liaoning Province, China. AB - The prevalence of and associated risk factors for isolated systolic hypertension, as defined by the Joint National Committee (JNC)-6 classification, were investigated in the rural population of Liaoning Province, China. A total of 45,925 people aged 35 years or older were examined in a cross-sectional study. Overall, the prevalence of isolated systolic hypertension was 10.6% (males 10.1%; females 11.2%). The prevalence of isolated systolic hypertension was positively correlated with age, gender, smoking status, alcohol intake, body mass index, salt intake and Mongolian race. This study showed that isolated systolic hypertension was very common in rural Chinese people and that many risk factors are linked with isolated systolic hypertension. PMID- 18380949 TI - Primary malignant myoepithelioma of the left maxillary sinus: a case report. AB - We describe the case of a 41-year-old woman who presented with a malignant myoepithelioma (MME) in her left maxillary sinus. Exploratory biopsy of the left maxillary sinus was performed and pathological examination demonstrated that the tumour was positive for calponin and cytokeratin 14, which are indicative of MME. Lateral rhinotomy and left total maxillectomy were undertaken and the patient received radiotherapy and chemotherapy post-surgery. Primary recurrence and metastasis to the left angle of the mandible occurred 9 months after the surgery. The patient died of cachexia 13 months after the surgery. PMID- 18380950 TI - Multiple primary primitive neuroectodermal tumours within the spinal epidural space with non-concurrent onset. AB - A case of multiple primary primitive neuroectodermal tumours (PNETs), which occurred at different levels of the spinal epidural space successively over a period of 8 months, is reported. A 24-year-old male, presenting with rapidly progressive paralysis, hyperthesia and a posterior epidural mass extending from T8 to T10 revealed by magnetic resonance imaging (MRI), exhibited a good recovery after initial emergency surgery. Lower back pain, chest pain and paralysis were subsequently reported. Spinal MRI in month 7 revealed a mass extending from T12 to L1 and another mass extending from T4 to T5 was detected epidurally in month 8. Additional operations were performed and radiotherapy was given. Pathological findings were consistent with PNETs and symptoms improved with treatment, particularly following each surgical excision. PMID- 18380951 TI - Chronic conditions that affect post-reproductive health. Cardiovascular disease and obesity. PMID- 18380952 TI - Osteoporosis prevention treatments under the spotlight. PMID- 18380953 TI - Cardiovascular disease in women. PMID- 18380954 TI - Red clover isoflavones are safe and well tolerated in women with a family history of breast cancer. AB - OBJECTIVE: To assess the safety and tolerability of a standardized 40 mg red clover isoflavone dietary supplement (Promensil, Novogen) in women with a family history of breast cancer to evaluate the feasibility of using the supplement for prevention of breast cancer in healthy women. STUDY DESIGN: Healthy women aged 35 70 years (n = 401) with at least one first-degree relative with breast cancer received red clover isoflavones or placebo for three years in a randomized, double-blind, placebo-controlled pilot trial. Participants were assessed clinically and blood samples taken for biochemical analysis every six months. In addition, study participants underwent mammography, bone density and transvaginal ultrasound (postmenopausal women only) once per year. RESULTS: No significant differences in breast density, endometrial thickness, serum cholesterol, follicle stimulating hormone levels and bone mineral density were detected between those taking red clover isoflavones and placebo. In postmenopausal women, some significant differences in bone marker levels were seen between active and placebo groups, at six months and at 12 months. The adverse event profile was similar across all red clover isoflavone and placebo groups. CONCLUSION: This three-year study supports the growing body of evidence that treatment with red clover isoflavones is safe and well tolerated in healthy women. Supplements containing red clover isoflavones did not adversely affect breast density, skeletal strength or cardiovascular status. In postmenopausal women, endometrial status was not adversely affected. The adverse event profile was similar between red clover isoflavones, and placebo and endocrine status did not differ. PMID- 18380955 TI - Gender differences in cardiovascular disease prevention. AB - Risk factors are generally shared between men and women with the major differences being hormonal. Nine modifiable risk factors account for over 90% of the risk of a coronary event in men and women--smoking, hypertension, hyperlipidaemia, diabetes, abdominal obesity, lack of exercise, alcohol excess, reduced intake of fruit and vegetables, and psychosocial issues. Approximately half the decline in deaths from coronary heart disease (CHD), between 1980 and 2000, can be attributed to a reduction in the major risk factors and the other half to the use of evidence-based management. As educational efforts to increase awareness of cardiovascular disease (not cancer) to be the leading cause of death and disability in women are also associated with preventative action, it is important that health-care professionals educate themselves about CHD in women and communicate with women themselves, so that women can come forward for advice and evaluation. PMID- 18380956 TI - Heart disease and gender in mass print media. AB - Heart disease is a major cause of death, disease and disability in the developed world for both men and women. Nevertheless, the evidence suggests that women are under-diagnosed both because they fail to visit the doctor with relevant symptoms and because doctors tend to dismiss the seriousness of women's symptoms of heart disease. This study examines the way that popular mass print media present the possible links between gender and heart disease. The findings suggest that the 'usual candidates' for heart disease are considered to be high achieving and active men for whom the 'heart attack' is sometimes seen as a 'badge of honour' and a symbol of their success. In contrast, women are less often seen as likely to succumb, but they are portrayed as if they are and ought to be worried about their husbands. Women's own bodies are described as so problematic as to be perhaps useless to diagnose, because they are so difficult to understand and treat. PMID- 18380957 TI - Metabolic syndrome and the menopause. AB - The metabolic syndrome consists of a combination of risk factors that include abdominal obesity, atherogenic dyslipidaemia, hypertension and insulin resistance. It increases the risk of cardiovascular disease and type 2 diabetes. The increased risk of cardiovascular disease is higher in women than in men. The first manifestation of metabolic syndrome may occur in pregnancy presenting as gestational diabetes or preeclampsia. Both conditions are associated with increased insulin resistance. Also metabolic syndrome is more common in polycystic ovarian syndrome. It has been suggested that there is a metabolic syndrome resulting from the menopause due to estrogen deficiency, as many of the risk factors are more prevalent in postmenopausal women. Also estrogen replacement improves insulin sensitivity and reduces the risk of diabetes. The key elements in managing the metabolic syndrome are weight reduction, increasing physical activity and diet modification. If blood pressure, lipid and glycaemic control are not achieved through these interventions then pharmacological therapy will be required. PMID- 18380958 TI - Quality of life in obese postmenopausal women. AB - The aim of this review was to identify the ways in which obesity affects the health-related quality of life (HRQoL) of postmenopausal women. This was considered important because a growing body of literature has identified obesity as a significant predictor for a poor psychological wellbeing and negative HRQoL, particularly in women, and because during the transition through the menopause women tend to accumulate more body weight. After searching eight electronic databases, only nine papers appeared meaningful. Although a meta-analysis was not possible, we found that a body mass index (BMI) > 30 kg/m2 was associated with a poor HRQoL in postmenopausal women; particularly in the areas associated with physical functioning, energy and vitality, and health perceptions. Thus, clinical management of obese postmenopausal women should focus on weight reduction and exercise in an attempt to improve wellbeing in these areas. However, the paucity of research, the different instruments chosen to measure HRQoL and the methodological limitations of the studies identified, prevented firm conclusions being made about whether the relationship between BMI and HRQoL is linear in postmenopausal women. Further research is needed to explore this relationship; particularly in comparison with underweight postmenopausal women and obese premenopausal women, where a few papers have identified these women as having a worse HRQoL than their obese postmenopausal counterparts. Other measures of central adiposity, including waist circumference and waist-hip ratio are recommended as useful supplemental measures to BMI in future studies. PMID- 18380959 TI - Obesity and gynaecological cancer. AB - Obesity is now considered to be a global epidemic. The problem of obesity has significant implications for the diagnosis and treatment of gynaecological cancer. The cancer most frequently associated with obesity is that of the endometrium. The risk of endometrial cancer is 2-3 times higher in overweight and obese women. Obesity also adversely affects survival in most studies. With regard to ovarian cancer the evidence is inconsistent. Obesity in young adulthood may be more important than that in later life. With regard to survival obesity has an adverse effect but not in early stage disease. Few data are available regarding cervical cancer and obesity. There is evidence that obesity is associated with adenocarcinoma rather than squamous carcinoma. Data on vulval cancer and obesity are scant. PMID- 18380960 TI - Management of Parkinson's disease in older women. AB - Elderly women with Parkinson's disease (PD) represent a specific patient population that may benefit from individualized treatment strategies. PD has been shown to occur approximately twice as often in men than in women, resulting in theories regarding estrogen being protective against the disease and as a potential treatment strategy. Given women's longer life expectancy, they are more likely to reach an age where antiparkinsonian medications are associated with side-effects. This paper will review medical and surgical treatments as well as the relationship of gender and age with respect to the management of PD. PMID- 18380961 TI - Primary prevention of coronary heart disease in women. AB - The British Menopause Society Council is committed to provide up-to-date authoritative reviews to aid health professionals to inform and advise women about key issues in postreproductive health. Coronary heart disease (CHD) is a leading cause of death in women. Observational studies have consistently shown estrogen to help prevent CHD in postmenopausal women. The large randomized controlled Women's Health Initiative (WHI) trial did not confirm these observational findings. However, further analyses of the WHI study as well as the observational Nurses' Health Study have now found that the timing of onset of hormone replacement therapy (HRT) use is important and that estrogen may have a protective role in CHD in women aged 50-59 years. This consensus statement will examine the evidence regarding HRT and non-estrogen therapies (lipid lowering agents, aspirin, antihypertensives, antidiabetic medications, selective estrogen receptor modulators [SERMs]) as well as diet, lifestyle and smoking cessation in the primary prevention of CHD in women. PMID- 18380962 TI - [Greetings for the new year]. PMID- 18380963 TI - [For clinical studies, do not forget the rules and regulations]. PMID- 18380965 TI - [Detection of periodontal pathogens in coronary atherosclerotic plaques]. AB - OBJECTIVE: To explore the possible correlation between coronary heart disease and periodontitis. METHODS: Subgingival plaque samples and coronary atherosclerotic plaques were harvested from a total of 31 patients with periodontitis who scheduled for coronary artery bypass surgery. The bacteria DNA was obtained from subgingival plaque samples and coronary atherosclerotic plaques using the chelex 100 method. The extracted DNA was examined using the polymerase chain reaction (PCR) technique. RESULTS: In coronary atherosclerotic plaques samples from the 31 patients, Porphyromonas gingivalis (Pg, 38.7%), Actinobacillus actinomycetemcomitans (Aa, 0%), Fusobacterium nucleatum (Fn, 22.6%), Prevotella intermedia (Pi, 12.9%), Bacteroides forsythus (Bf, 38.7%) were detected. The concordant presence of the same periodontal bacteria DNA in subgingival plaques and in coronary atherosclerotic plaques in the same patient was Pg 5 (16.1%), Aa 0 (0%), Pi 2 (6.5%), Fn 4 (12.9%) and Bf 8 (25.8%). CONCLUSIONS: The presence of periodontal bacteria DNA in coronary atherosclerotic plaques could indicate that periodontal pathogenic bacteria may play a role in the coronary heart disease process. PMID- 18380966 TI - [Effect of fibrinogen on the secretion of interleukin-1beta and - 8 by polymorphonuclear leukocytes]. AB - OBJECTIVE: To investigate the effect of one of the acute-phase proteins, fibrinogen, on the release of IL-1beta and -8 by human peripheral polymorphonuclear leukocytes (PMN) and the possible role of fibrinogen during the destruction of periodontium. METHODS: Peripheral PMN were isolated by discontinuous density gradient centrifuging technique. The freshly isolated PMN were suspended in Hank's balanced saline solution (1 x 10(9)/L) supplemented with 0.5% BSA and 0.1% glucose. The levels of IL-1beta and -8 in the supernatants produced by cultured cells upon the addition of human fibrinogen at different concentrations were measured by ELISA technique. RESULTS: Incubated with human fibrinogen at 2 g/L or 10 g/L for different time periods, human peripheral PMN released significantly greater amount of IL-1beta [(10.41 +/- 0.37) - (35.86 +/- 0.30) ng/L or (22.81 +/- 0.45) - (57.77 +/- 2.08) ng/L] and IL-8 [(93.90 +/- 13.95) - (2045.66 +/- 53.03) ng/L or (115.02 +/- 10.61) - (3858.69 +/- 25.65) ng/L] than PMN without the stimulation of fibrinogen (IL-1beta, P < 0.001, and IL 8, P < or = 0.016). The higher concentration of fibrinogen or the longer treatment time, the higher levels of IL-1beta and -8 were released by PMN (P < 0.001). CONCLUSIONS: Fibrinogen induced the secretion of pro-inflammatory cytokines IL-1beta and -8 by PMN and may be involved in magnification of the inflammatory response of periodontium and bone resorption. PMID- 18380967 TI - [Effect of advanced glycation end products on the human gingival fibroblast proliferation and type I collagen synthesis]. AB - OBJECTIVE: To apply the synthesized advanced glycation end products (AGE) to the cultured human gingival fibroblast (HGF) in vitro and then to investigate the effects of AGE on the HGF proliferation and type I collagen synthesis and the potential impact of AGE in the repair of periodontium and its molecular mechanism in diabetes-associated periodontitis. METHODS: The HGF was obtained from explants of human healthy gingival tissues by using tissue-explant technique. The AGE was prepared and then added to the culture media, its effect on HGF proliferation at different time duration was examined with MTT colorimetric assay. The type I collagen concentrations in cell culture supernatants and intracellular proteins were detected by ELISA, and the type I collagen mRNA expression of HGF was analyzed by real-time RT-PCR. RESULTS: 200 mg/L AGE decreased the A value (P < 0.05) and changed the HGF shape. Incubation of HGF with AGE for 72 hours, the quantities of type I collagen were reduced (P < 0.05), and the expression of type I collagen mRNA was down-regulated (P < 0.05). CONCLUSIONS: The AGE inhibited the HGF proliferation, decreased the synthesis of type I collagen and down-regulated the expression of type I collagen mRNA, impairing the repair of periodontium. PMID- 18380968 TI - [Treatment of maxillary protrusion in adult with self-tapping micro-screw implants]. AB - OBJECTIVE: To investigate the position change of upper molars and incisors in order to evaluate the stability of posterior anchorage with the application of micro-screw implant. METHODS: Eight adult patients with severe maxillary protrusion were included. Upper first premolars were extracted and upper posterior anchorage was reinforced with micro-screw implant in all patients. Cephalometric and cast analyses were carried out to record the position change of molar and micro-screw. RESULTS: During treatment the micro-screw implants kept stable in sagittal and vertical plane. Neither the mesial-distal movement nor the tipping of the upper molars during the treatment was statistically significant (P > 0.05). The edge of upper incisors was retracted by 6.86 mm and the tipping was reduced by 18.03 degrees . The center of resistance was intruded by 3.28 mm on average. Significant change was observed (P < 0.01). CONCLUSIONS: Micro-screw implant could provide good anchorage control in the orthodontic treatment. PMID- 18380969 TI - [Effect of simvastatin on bone morphogenetic protein-2 expression in the periodontal tissue after rat tooth movement]. AB - OBJECTIVE: To investigate the effect of systematic administration of simvastatin on the bone morphogenetic protein-2 (BMP-2) expression in the periodontal tissue after rat tooth movement and on the relapse of tooth movement. METHODS: Orthodontic tooth movement of upper first molar was performed in 32 rats with coil spring for 21 days. The 32 rats were randomly allocated into 4 groups: negative control group (isotonic saline) and three experimental groups (2.5 mg x kg(-1), 5.0 mg x kg(-1) and 10.0 mg x kg(-1)). The simvastatin started to be administered to the experimental groups 1 day before appliances were removed, and once a day there after for 4 weeks. The negative control group received the isotonic saline only. The interdental distance between the first and second maxillary molars were measured, when appliances were removed, and 1 week and 4 weeks after that. After the rats were sacrificed, sections of first maxillary molar and periodontal tissue were studied by immunohistochemistry. RESULTS: The number and percentage of relapse was lower in the three experimental groups than in the negative control group (P < 0.05, P < 0.01). The lower dose was given, the less relapse there was, with the lowerest dose resulting in lowest percentage of relapse (26.81% and 53.38%). BMP-2 expression in experimental groups was higher than in the negative control group, with the lowerest dose group showing the highest expression (P < 0.001). The BMP-2 expression on the tension side was slightly stronger than that on the compression side (P > 0.05). CONCLUSIONS: Systemic administration of simvastatin could decrease the extent of relapse of the orthodontic-moved tooth in rat, and the lower-dose of simvastatin seemed more effective. The possible mechanism for this may be that simvastatin functions by increasing the expression of BMP-2 in the periodontal tissue, accelerating the osteoblast activity and promoting bone formation. PMID- 18380970 TI - [Evaluation of patients' temporomandibular joint function after mandible reconstruction with free fibula flap]. AB - OBJECTIVE: To evaluate the temporomandibular joint (TMJ) function after mandible reconstruction with free fibula flap and with different condyle treatment. METHODS: Forty-one cases of mandible and condyle defects following benign tumor resection were reconstructed with fibula free flap, with the condyle being reconstructed by three methods: placement of the distal end of the fibula flap into the glenoid fossa, the fibula serving as the condyle (24 cases); attachment of the resected condyle as a nonvascularized transplant to the end of the fibula flap (5 cases); condyle preservation plus vascularized fibular reconstruction (12 cases). The postoperative TMJ function was evaluated by Fricton function index. RESULTS: No TMJ ankylosis occurred in all patients. There were no significant differences in aesthetics, deglutition and speech function among the three groups; but there were significant differences in TMJ function index (DI, CMI) among different condyle treatments. Condylar preservation showed better TMJ function than fibula used as a substitute for condyle. CONCLUSIONS: Preservation of condyle during free fibula mandibular reconstruction can improve patients' TMJ function. The location and shape of fibular condyle were important factors that could influence the outcome the condyle reconstruction. PMID- 18380971 TI - [Influence of chronic unpredictable emotional stress on the ultrastructures of temporomandibular joint in rats]. AB - OBJECTIVE: To study the influence of emotional stress on the ultrastructures of temporomandibular joint in rats. METHODS: Sixteen male Wistar rats, aged 12 weeks, were divided into 2 groups randomly, experimental group and control group. Animal model of chronic unpredictable stress was established in the experimental group. After 6 weeks, all rats in both groups were killed and their condyles and articular discs were dissected, taken out and observed by scanning electron microscope. RESULTS: Under scanning electron microscope, the surface gelatum of the condyles in experimental group were cracked, and collagen fibrils in disorder and broke, and deep layer collagen fibrils were exposed. The articular discs' surface gelatum were still integrated, but the gyri dispose were irregular. In control group, there were no such changes in condyles and articular discs. CONCLUSIONS: The long-term emotional chronic unpredictable stress could induce the injuries of temporomandibular joint in rats in ultrastructure level. PMID- 18380972 TI - [Electrosonographic analysis of sounds emanating from temporomandibular joint without symptom]. AB - OBJECTIVE: To study the characteristic of joint sounds from temporomandibular joint (TMJ) without symptom during mandible movement. METHODS: Joint sounds of 42 asymptomatic TMJ were recorded and analyzed with K6-I system during mandible movement, the waveform, frequency range, peak frequency and amplitude of joint sounds electrosonogram were studied. RESULTS: Joint sounds were recorded in 25 joints of the 42 joints, 4 kinds of joint sounds were found according to the waveform, frequency range, peak frequency and amplitude of the electrosonography. CONCLUSIONS: Joint sounds can be recorded in asymptomatic TMJ during mandible movement, 4 kinds of joints sounds were found according to the electrosonographic characteristics. PMID- 18380973 TI - [Detection of quorum-sensing pathway and construction of luxS gene allelic exchange plasmid of Streptococcus mutans]. AB - OBJECTIVE: To detect the AI-2 quorum-sensing pathway and construct the luxS g-ene allelic exchange plasmid of Streptococcus mutans. METHODS: To detect AI-2 pathway in Streptococcus mutans, the Vibrio harveyi BB170 was used as reporter strain. The PCR fragments of the upstream and downstream regions of luxS and the Erythromycin resistance gene were amplified with the primers respectively, and these fragments were ligated into pUC19 vector with double endonuclease reaction sequentially, the ligated DNAs were transformed into Escherichia coli DH5alpha, then the reconstructed plasmids were isolated and identified by restricted endonuclease digestions. RESULTS: Streptococcus mutans Ingbritt C could induce luminescence of BB170, suggesting the presence of AI-2 quorum sensing pathway in Streptococcus mutans, and such stimulatory activity was maximal at the mid-log growth phase. The recombinant plasmid pUCluxKO was digested by PstI-BamHI, and the digest product were 1000 bp and 5000 bp. When the pUCluxKO was digested by BamHI-KpnI, the digest product were 1500 bp and 4500 bp. While it was digested by KpnI-EcoRI, the digest product were 1000 bp and 5000 bp. All PCR product was in a single belt respectively. CONCLUSIONS: The recombinant plasmid was cloned effectively and can be used in the construction of S.mutans luxS mutant. PMID- 18380974 TI - [Shaping ability of two nickel-titanium rotary systems in simulated S-shaped canals]. AB - OBJECTIVE: To evaluate the shaping ability of two nickel-titanium rotary systems (ProTaper and Hero642) in simulated S-shaped canals. METHODS: Thirty simulated S shaped canals were randomly divided into three groups and prepared by ProTaper, Hero642, ProTaper combined with Hero642 respectively. All the canals were scanned before and after instrumentation, and the amount of material removed in the inner and outer wall and the canal width after instrumentation were measured with a computer image analysis program. RESULTS: There was significant difference in the amount of material removed at the inner side of apical curvature and outer side of apex between ProTaper combined with Hero642 and ProTaper files (P < 0.05) at the same tip size. The inner and outer wall of the canals were evenly prepared by ProTaper combined with Hero642, and the taper of canals were better than those prepared by Hero642. CONCLUSIONS: ProTaper combined with Hero 642 had better shaping ability to maintain the original shape and could create good taper canals in the simulated S-shaped canal model. PMID- 18380975 TI - [Clinical evaluation of composite inlays in defective molars]. AB - OBJECTIVE: To evaluate clinical effect of composite inlays in the defective molars. METHODS: A total of 200 defective molars from 163 patients were divided into two groups, including 100 molars of each. One group was restored with the direct composite inlays and another group with the traditional composite fillings. All the restorations were evaluated in oral cavity after 6-month and 5 year filling or insertion with United States public health service criterions. The data were analyzed using SPSS 11.0 software with the chi-square test. The significance level was set at 5%. RESULTS: In clinical service for 6 months, the successful rate of composite inlays was 91.8% (90/98) and the corresponding figure for traditional composite fillings was 91.8% (89/97), but there was no statistically significant difference (P > 0.05). In clinical service for 5 years, the successful rate of composite inlay was 87.9% (80/91), the corresponding figure for the traditional composite fillings being 67.4% (60/89) and the difference was statistically significant (P < 0.05). CONCLUSIONS: In clinical, the defective molars can be well restored with the direct composite inlays. Especially in the long-term clinical service, the composite inlays show significant superiority over the traditional composite fillings. PMID- 18380977 TI - [Effect of simulating static magnetic field of magnetic attachment on osteoblastic morphology and surface ultrastructure]. AB - OBJECTIVE: To investigate effects of the static magnetic field (SMF) generated by dental magnetic attachment on osteoblastic morphology and surface ultrastructure. METHODS: The in vitro cultured rat osteoblasts were exposed continuously to 12.5 mT, 125 mT, and 250 mT static magnetic fields for 1, 3, 5, and 7 days. After exposed in SMF, osteoblasts were observed under a phase contrast microscope, and then HE stained and observed under a light microscope. In addition, the cells were observed under a scanning electron microscope (SEM). RESULTS: By continuous exposure, the different intensities of SMF exposure did not change the vital osteoblast growth pattern or distribution. The SEM photos showed that there were certain changes in cellular microstructures for osteoblasts after exposed to 12.5 mT for 5 to 7 days, as well as 125 mT and 250 mT for 3 to 7 days. The more exposure time increased, the more microvesicles on the surfaces of cells were observed. CONCLUSIONS: Continuous SMF-stimulation could not affect the shape, distribution, and growth pattern of osteoblasts. The SMF of magnetic attachments could lead to certain changes in surface ultrastructures of osteoblasts in this study. PMID- 18380978 TI - [Influence of implant-abutment connection mode on stress distribution in peri implant bone]. AB - OBJECTIVE: To investigate the influence of butt joint connection and platform switching design of implant-abutment connection on the stress distribution in peri-implant bone. METHODS: Three-dimensional finite element models of implant supported mandibular first molar with different implant-abutment connections were computed by COSMOSM 2.85. Traditional butt joint connection was used in model A and platform switching design in model B. Loading conditions were a vertical load of 200 N and inclined load of the same magnitude at 45 degrees to the vertical axis of the implant. Stress distribution in peri-implant bone and von Mises stresses at the same point of buccal and lingual implant-bone interfaces in two models were compared. RESULTS: Stresses concentrated in peri-implant cortical bone at the neck of implants on the buccal and lingual sides; maximum von Mises stresses under inclined load were higher than those under vertical load. Maximum von Mises stress in bone was 11.61 MPa in model A and 7.15 MPa in model B under vertical load, and 22.07 MPa in model A and 11.87 MPa in model B under inclined load respectively. Von Mises stresses decreased as the distance from implant abutment junction increased and the most obvious change occurred at the interface between cortical bone and spongy bone. Von Mises stresses at the same points of buccal and lingual implant-bone interfaces in model A were higher than those in model B. CONCLUSIONS: Compared with butt joint connection, platform switching design improved the stress distribution and decreased the maximum stresses in peri-implant bone around implant cervix. PMID- 18380979 TI - [Silver-ion release and particle distribution of denture base resin containing nanometer-sized silver-supported antimicrobial agent]. AB - OBJECTIVE: To investigate the distribution of antimicrobial agent STR-1 of nanometer level which was incorporated with ball-grinding method in the polymethylmethacrylate (PMMA) denture base, and to study the release mode of silver ions from the base. METHODS: The distribution of the antimicrobial agent in the PMMA denture base containing STR-1 at concentrations of 0 g/L, 5 g/L, and 10 g/L was examined with scanning electronic microscopy. Then, PMMA resin bases containing STR-1 at the three concentrations were respectively immersed in artificial saliva at 37 degrees C for 54 days. The release of silver ions from the resin bases was surveyed with inductively coupled plasma-mass spectroscopy (ICP-MS) every 24 hours. RESULTS: The antimicrobial agent incorporated by ball grinding method was even-distributed with individual particles of nanometer level in the PMMA resin base. The release of silver ions from the PMMA resin with antimicrobial agent was extremely slow during the test, a very small fraction of the silver ions released. At the beginning of the test, the release speed was extremely slow, the speed increased rapidly in the middle of the test, and at the end of the test, the speed returned to slow and steady. The cumulative release curve of silver ions was of "S" type. CONCLUSIONS: STR-1 can be even-distributed in the denture base, and the silver ions release from the base with extremely slow speed. It also indicates that biological safety and long-term antimicrobial efficacy of denture base containing silver-supported antimicrobial agents of nanometer level are possibly obtained based on their slow release of silver ions. PMID- 18380980 TI - [Biological transport of tetracycline hydrochloride by human periodontal ligament fibroblasts]. AB - OBJECTIVE: To investigate biological transport of tetracycline hydrochloride by human periodontal ligament fibroblasts (HPDLF) for verifying the hypothesis of delivering medicine to the periodontium and whole body through the root canal. METHODS: HPDLF and MC3T3-E1 cells were incubated in antibiotics solutions. The intracellular antibiotics contents were measured by high performance liquid chromatography (HPLC) and the cell total protein was measured by bradford protein assay. RESULTS: The intracellular contents increased with incubation time. The extracellular medicine concentration had effect on the intracellular contents. CONCLUSIONS: Tetracycline hydrochloride can be transported into HPDLF with incubation and this transport is time-dependent and concentration-dependent. PMID- 18380982 TI - Mushroom-like gastric tumor. PMID- 18380983 TI - Critical glucose control: the devil is in the details. PMID- 18380984 TI - Thrombophilia screening: little role for the JAK2V617F mutation. PMID- 18380985 TI - Accuracy of bedside glucometry in critically ill patients: influence of clinical characteristics and perfusion index. AB - OBJECTIVES: To determine the accuracy of bedside glucose strip assay on capillary blood and on whole blood and to identify factors predictive of discrepancies with the laboratory method. PATIENTS AND METHODS: We conducted a prospective 3-month (July 1-September 30, 2003) study in 85 consecutive patients who required blood glucose monitoring. Values obtained with a glucose test strip on capillary blood and on whole blood were compared with those obtained in the laboratory during serial blood sampling (up to 4 samples per patient). The test strip values were considered to disagree significantly with the laboratory values when the difference exceeded 20%. Clinical and biological parameters and the perfusion index, based on percutaneous oxygen saturation monitoring, were recorded when each sample was obtained. RESULTS: Capillary glucose values conflicted with laboratory reference values in 15% of samples. A low perfusion index was predictive of conflicting values (P=.04). Seven percent of values obtained with glucose strip on whole-blood samples conflicted with laboratory reference values; factors associated with these discrepancies were mean arterial hypotension (P=.007) and generalized mottling (P=.04). CONCLUSION: Bedside blood glucose values must be interpreted with care in critically ill patients. A low perfusion index, reflecting peripheral hypoperfusion, is associated with poor glucose strip performance. Bedside measurements in whole blood seem to be most reliable, except in patients with arterial hypotension and generalized mottling. PMID- 18380986 TI - Hyperglycemia in patients undergoing cerebral aneurysm surgery: its association with long-term gross neurologic and neuropsychological function. AB - OBJECTIVE: To evaluate whether elevated intraoperative blood glucose concentrations are associated with an increased risk of long-term neurologic dysfunction in patients at risk for ischemic brain injury. PATIENTS AND METHODS: Data from 1000 patients were retrieved from the Intraoperative Hypothermia for Aneurysm Surgery Trial database. All patients were recruited between February 2000 and April 2003, and underwent surgery for aneurysm clipping within 14 days of subarachnoid hemorrhage. Gross neurologic and neuropsychological function was evaluated at 3 months after surgery using certified observers and standardized assessment instruments. Intraoperative blood glucose concentrations, measured once when the aneurysm clip was placed, were correlated with neurologic outcome using both univariable and multivariable logistic regression analyses. RESULTS: Blood glucose concentrations at the time of aneurysm clipping ranged from 59 to 331 mg/dL. At 3 months after surgery, those with blood glucose concentrations of 129 mg/dL or more (upper 2 quartiles) were more likely to have impaired cognition (P=.03). Those with glucose concentrations of 152 mg/dL or more (upper quartile) were more likely to experience deficits in gross neurologic function assessed by the National Institutes of Health Stroke Scale (P<.05), but not other scoring scales. Length of stay in intensive care units was longer in those with glucose concentrations of 129 mg/dL or more, but there was no difference among glucose groups in the duration of overall hospital stay or the fraction of patients discharged to home. CONCLUSION: In patients at high risk for ischemic brain injury, intraoperative hyperglycemia, of a magnitude commonly encountered clinically, was associated with long-term changes in cognition and gross neurologic function. PMID- 18380988 TI - Hysteroscopic sterilization in women with severe cardiac disease: experience at a tertiary center. AB - OBJECTIVE: To determine the feasibility and effectiveness of hysteroscopic sterilization as a minimally invasive sterilization method for women with high risk cardiac disease. PARTICIPANTS AND METHODS: In a retrospective cohort study, 18 women with high-risk cardiac conditions that strictly contraindicated pregnancy were compared with a reference cohort of 157 women without cardiac disease. All underwent microinsert hysteroscopic sterilization at Mayo Clinic from January 2003 through February 2007. End points included successful placement, fallopian tube patency determined by hysterosalpingogram 3 months after the procedure, and pregnancy status. RESULTS: Women in the cardiac cohort were younger than those in the reference cohort (median age, 25 vs 39 years; P<.001), had lower parity (median, 0 vs 2; P<.001), and had a higher proportion of patients categorized as American Society of Anesthesiologists' physical status 3 (severe systemic disease) and physical status 4 (systemic disease that is a constant threat to life) (83% vs 6%; P<.001). No significant differences were noted for use of general anesthesia (17% vs 27%; P=.41), successful bilateral device placement (100% vs 95%; P>.99), postoperative pain score (median, 0 for both groups; P=.87), or length of hospitalization (median, 6 vs 6 hours; P=.63). No intraoperative complications occurred. Follow-up hysterosalpingography showed high tubal occlusion rates in both cohorts (100% cardiac; 98% reference; P>.99). No pregnancies occurred during a median follow-up period of 20 months (interquartile range, 8-33 months). CONCLUSION: For women with cardiac disease and strict contraindications for pregnancy, microinsert hysteroscopic sterilization provided minimally invasive, permanent, and reliable contraception. PMID- 18380987 TI - Effect of perioperative insulin infusion on surgical morbidity and mortality: systematic review and meta-analysis of randomized trials.7. AB - OBJECTIVE: To conduct a systematic review and meta-analysis of randomized controlled trials (RCTs) to evaluate the effect of perioperative insulin infusion on outcomes important to patients. PATIENTS AND METHODS: We used 6 search strategies including an electronic database search of MEDLINE, EMBASE, and Cochrane CENTRAL, from their inception up to May 1, 2006, and included RCTs of perioperative insulin infusion (with or without glucose targets) measuring outcomes in patients undergoing any surgery. Pairs of reviewers working independently assessed the methodological quality and characteristics of included trials and abstracted data on perioperative outcomes (ie, outcomes that occurred during hospitalization or within 30 days of surgery). RESULTS: We identified 34 eligible trials. In the 14 trials that assessed mortality, there were 68 deaths among 2192 patients randomized to insulin infusion compared with 98 deaths among 2163 patients randomized to control therapy (random-effects pooled relative risk, 0.69; 95% confidence interval [CI], 0.51-0.94; 99% CI, 0.46-1.04; I2, 0%; 95% CI, 0.0%-47.4%). Hypoglycemia increased in the intensively treated group (20 trials, 119/1470 patients in insulin infusion vs 48/1476 patients in control group; relative risk, 2.07; 95% CI, 1.29-3.32; 99% CI, 1.09-3.88; I2, 31.5%; 95% CI, 0.0%-59.0%). No significant effect was seen in any other outcomes. The available mortality data represent only 40% of the optimal information size required to reliably detect a plausible treatment effect; potential methodological and reporting biases weaken inferences. CONCLUSION: Perioperative insulin infusion may reduce mortality but increases hypoglycemia in patients who are undergoing surgery; however, mortality results require confirmation in large and rigorous RCTs. PMID- 18380989 TI - Prevalence of delayed clinician response to elevated prostate-specific antigen values. AB - OBJECTIVE: To assess the frequency of delayed response to an abnormal prostate specific antigen (PSA) value. PATIENTS AND METHODS: Retrospective review of prostate cancer cases diagnosed between January 1, 2000, and December 31, 2005, in a rural Department of Veterans Affairs health care system serving 44,000 veterans across 2 states. Clinician response was defined as a reference to the elevated PSA result in clinical notes, orders for further evaluation, treatment of presumed prostatitis, or a urology visit or referral. Delay was measured as days between an abnormal PSA result and clinician response. RESULTS: We identified 327 men who met inclusion criteria with an abnormal PSA value before prostate cancer diagnosis. At first PSA elevation, median age was 64 years; 94% were younger than 75 years. Of the 327 men, 253 (77.4%) had a timely (< or =30 days) response to an abnormal PSA value; 23 (7.0%) had between 31 and 180 days; 24 (7.3%), between 181 and 360 days; and 27 (8.3%), more than 360 days between an abnormal PSA measurement and clinician response. The delayed group had nearly an additional year's (309 days) lapse before completed urologic consultation and prostate gland biopsy (313 days) as compared with the timely group. The presence of urologic symptoms, abnormal results from rectal examination, higher PSA values, and higher PSA velocity (P<.05) were associated with timely clinician response to an abnormal PSA measurement. CONCLUSION: In a cohort of men with prostate cancer and an antecedent abnormal PSA value, 15.6% had more than 180 days between an abnormal PSA measurement and clinician response. These findings add to the growing literature demonstrating that missed results occur more frequently than is generally appreciated. Improved systems for clinical data management are needed. PMID- 18380990 TI - Associations of conference attendance with internal medicine in-training examination scores. AB - OBJECTIVE: To examine the association of medical knowledge acquisition with attendance at specific conference types during internal medicine residency. PARTICIPANTS AND METHODS: Attendance at residency core curriculum, morbidity and mortality, and medical grand rounds conferences was tracked for 195 residents who took the Internal Medicine In-Training Examination (IM-ITE) 421 times while attending the Internal Medicine Residency Program at Mayo Clinic's site in Rochester, MN, between October 1, 1999, and October 1, 2003. Random effects modeling was used to adjust for variables known or hypothesized to be associated with IM-ITE scores and allowed discrimination of effects of individual conferences on IM-ITE scores. RESULTS: Total conference attendance was significantly associated with increased IM-ITE scores, with cohort mean increase of 2.30% (95% confidence interval [CI], 1.24% to 3.36%; P<.001). Core curriculum conferences accounted for much more of this association than either medical grand rounds or morbidity and mortality, with cohort mean increases of 2.05% (95% CI, 0.13% to 3.90%; P=.04), 0.24% (95% CI, -0.56% to 1.03%; P=.61), and 0.03% (95% CI, -1.69% to 1.69%; P=.97), respectively. CONCLUSION: Conference attendance is associated with improved knowledge acquisition as measured by improved performance on IM-ITE. This finding does not appear to apply to all conferences at our institution but rather is specifically attributable to attendance at the core curriculum series. PMID- 18380991 TI - JAK2V617F mutation screening as part of the hypercoagulable work-up in the absence of splanchnic venous thrombosis or overt myeloproliferative neoplasm: assessment of value in a series of 664 consecutive patients. AB - The JAK2V617F mutation is recurrent in polycythemia vera and essential thrombocythemia, which are myeloproliferative neoplasms (MPNs) frequently associated with arterial and/or venous thromboembolism. More recently, the JAK2V617F mutation has been identified as a surrogate marker for subclinical or "occult" clonal myeloproliferation in patients with splanchnic venous thrombosis. However, information is limited regarding JAK2V617F-associated thrombosis outside the splanchnic district in patients without overt MPN. To address this issue, we retrospectively studied a consecutive series of 664 such patients who experienced thrombotic events characteristic of an MPN (500 with venous thromboembolism, 136 with stroke, and 28 with myocardial infarction at a young age). The JAK2V617F mutation was detected in only 6 (<1.0%) patients (5 with recurrent venous thromboembolism and 1 with stroke), and the mutant allele burden was low in all instances (range, 2.2%-7.5%). None of these 6 patients developed either overt MPN or recurrent thrombosis after a median follow-up of 40 months. We conclude that the prevalence of the JAK2V617F mutation in patients with nonsplanchnic venous thrombosis in the absence of MPN is too low to warrant mutation screening as part of the hypercoagulable work-up. Our study also suggests that the natural history of a JAK2V617F-positive "occult" MPN might be different from that of a typical MPN. PMID- 18380992 TI - Gut microbiota and its possible relationship with obesity. AB - Obesity results from alterations in the body's regulation of energy intake, expenditure, and storage. Recent evidence, primarily from investigations in animal models, suggests that the gut microbiota affects nutrient acquisition and energy regulation. Its composition has also been shown to differ in lean vs obese animals and humans. In this article, we review the published evidence supporting the potential role of the gut microbiota in the development of obesity and explore the role that modifying the gut microbiota may play in its future treatment. Evidence suggests that the metabolic activities of the gut microbiota facilitate the extraction of calories from ingested dietary substances and help to store these calories in host adipose tissue for later use. Furthermore, the gut bacterial flora of obese mice and humans include fewer Bacteroidetes and correspondingly more Firmicutes than that of their lean counterparts, suggesting that differences in caloric extraction of ingested food substances may be due to the composition of the gut microbiota. Bacterial lipopolysaccharide derived from the intestinal microbiota may act as a triggering factor linking inflammation to high-fat diet-induced metabolic syndrome. Interactions among microorganisms in the gut appear to have an important role in host energy homeostasis, with hydrogen-oxidizing methanogens enhancing the metabolism of fermentative bacteria. Existing evidence warrants further investigation of the microbial ecology of the human gut and points to modification of the gut microbiota as one means to treat people who are over-weight or obese. PMID- 18380993 TI - Effects of niacin on glucose control in patients with dyslipidemia. AB - Niacin (nicotinic acid), the most effective available pharmacotherapy for increasing high-density lipoprotein cholesterol, also lowers triglycerides and hence may be useful, alone or in combination with hydroxymethylglutaryl coenzyme A reductase inhibitors (statins), to offset residual cardiovascular risk in patients with mixed or diabetic dyslipidemia. We conducted a review of published consensus guidelines since 2000 and an English-language PubMed search of prospective, randomized controlled trials and open-label studies from January 1, 1990, through December 31, 2007, concerning the effects of niacin, alone or in combination with statins, on glycemic regulation in dyslipidemic patients (with or without diabetes mellitus). For search terms, we used the title words niacin or nicotinic acid and key words including diabetes, diabetic, dyslipidemia, glucose, glycemic, HbA1c, hemoglobin, hyperglycemia, human, insulin, postprandial, and safety. Retrospective and observational studies, case reports, and case studies were excluded. On the basis of our analysis, the effects of niacin (< or =2.5 g/d), alone or in combination with statins, on fasting glucose (an increase of 4%-5%) and hemoglobin A1c levels (an increase of < or =0.3%) are modest, transient or reversible, and typically amenable to adjustments in oral hypoglycemic regimens without discontinuing niacin. Niacin therapy was infrequently associated with incident diabetes or the need for new insulin prescriptions. Studies showed important clinical benefits of niacin or niacin statin regimens despite modest effects on glucose control. On a population basis, significant reductions in incidences of cardiovascular events and the degree of atherosclerotic progression associated with long-term niacin (or niacin-statin) therapy in patients with diabetic dyslipidemia outweigh the typically mild effects of this therapy on glycemic regulation. Consensus guidelines recommend monitoring glycemic control after initiating niacin treatment or increasing its dosage. PMID- 18380994 TI - Treatment of tobacco dependence. AB - Cigarette smoking continues to cause substantial death and disability, but more than 1 in 5 adults smoke despite the desire among most smokers to stop and the availability of effective treatments. A systematic process to identify all smokers is crucial. Because tobacco dependence is characterized by relapses and remissions, clinicians should be ready to engage smokers and reengage relapsed smokers with options for new medication strategies and additional counseling resources. PMID- 18380995 TI - 55-year-old woman with menopausal symptoms and a family history of breast cancer. PMID- 18380996 TI - Head and neck cancer: changing epidemiology, diagnosis, and treatment. AB - Head and neck cancers account for less than 5% of all cancers and for less than 3% of all cancer deaths in the United States. The populations at risk for head and neck cancers are those who have a long-standing history of smoking and alcohol use. More recently, the incidence of oropharyngeal cancer in younger populations has been increasing and is associated with exposure to the human papillomavirus. This subset of patients appears to have a better overall prognosis and to respond better to treatment. This review is limited to head and neck cancers of squamous cell histology, which constitute more than 90% of head and neck cancers. Because treatment of head and neck cancers is complex and involves multiple modalities, a multidisciplinary approach is needed. This review focuses on the goal of organ preservation and postoperative treatment of high risk patients with the concurrent use of chemotherapy and radiation therapy. This review also highlights recent advances in treatment using molecularly targeted therapies, specifically the role of inhibitors of the epidermal growth factor receptor in locally advanced and recurrent/metastatic squamous cell cancer of the head and neck. Studies in the English language were identified by searching the MEDLINE, EMBASE database (1980-2007) using the search terms head and neck, squamous cell, carcinoma, chemotherapy, radiation, human papillomavirus, epidermal growth factor receptor, and targeted therapy. PMID- 18380997 TI - Carbamazepine as the only effective treatment in a 52-year-old man with trigeminal trophic syndrome. AB - Trigeminal trophic syndrome is a rare condition resulting from self-manipulation of the skin after a peripheral or central injury to the trigeminal system. The syndrome consists of a classic triad of anesthesia, paresthesias, and secondary persistent or recurrent facial ulcerations. The most common causes include destruction of the trigeminal ganglion, rhizotomy, and stroke. We describe a patient who developed the syndrome as a sequel to brainstem infarction and trigeminal neuropathy. Whereas a-lipoic acid and gabapentin were ineffective, a remarkable benefit was achieved by administering carbamazepine (200 mg 3 times a day), which influences both neuropathic and behavioral factors in this rare syndrome. Our experience with the presented case, together with the scarce information in the literature, indicates that carbamazepine should be the first treatment option for trigeminal trophic syndrome. PMID- 18380998 TI - Reconciling new organ donation laws with patient's autonomy and rights to self determination of end-of-life medical care. PMID- 18380999 TI - Immaculate conception. PMID- 18381000 TI - [Vitamin D: beyond treatment of osteoporosis]. PMID- 18381001 TI - [Ankle-brachial index testing is particularly indicated in patients with metabolic syndrome but without known arterial disease]. AB - INTRODUCTION: The objective was to investigate the prevalence of metabolic syndrome (MetS), its cardiovascular disease (CVD) risk and its association with peripheral arterial disease, defined by a low ankle-brachial index (ABI) (< 0.90), in primary prevention. MATERIAL AND METHODS: Cross-sectional study performed in the primary care general population, aged 50-79 years. CVD risk and ABI were measured in persons without known arterial disease with MetS and in a control group. MetS was defined by Adult Treatment Panel III. CVD risk was estimated using the Systematic Coronary Risk Evaluation (SCORE). RESULTS: A total of 581 subjects were recruited. The prevalence of the metabolic syndrome was 30.8% (95% confidence interval [CI], 27.0-34.5). For the CVD risk and ABI studies 217 individuals were included (138 with MetS and 49 with diabetes). Average CVD risk was high (5%) on patients with MetS and twice as much as those without MetS. ABI was low (< 0.90) on 15 subjects [6.9% (95% CI), 3.5-10.2], with higher frequency in the MetS group: 14 patients (10.1%) vs 1 patient (1.3%). The frequency of low ABI in patients with and without diabetes was 18.3% and 3.6% respectively. Low ABI was associated to MetS, diabetes, inactivity, high risk of SCORE and less alcohol habit. In the multivariate analysis, the odds ratio for the association MetS/low ABI was 14.7 (95% CI, 1.7-123.6). CONCLUSION: MetS is related to asymptomatic peripheral arterial disease. Thus, the measure of ABI in those patients with this syndrome is recommended. PMID- 18381002 TI - [Specialized home care of medical diseases in an urban tertiary university hospital. Coordination between the medical services of the hospital and the primary health care]. AB - INTRODUCTION: The experience of an urban Tertiary University Hospital in the design and implementation of Hospital at Home Program (HaHP) integrated in a Department of Internal Medicine and highly coordinated with Medical Services of the Hospital and the Primary Health Care, that contemplates the like main objectives of the promotion of specialized home care medical diseases and the improvement of the coordination with the primary health care. PATIENTS AND METHOD: Systematic collection in all the patients admitted between April 2006 and March 2007 in the HaHP of the following variables: age, gender, service of origin, main diagnosis, Barthel and Charlson index, number of visits per day to doctors and nurses, destination on discharge and medical team. The descriptive statistical analysis was made in April 2007. The results are presented globally and differentiated by teams (internal medicine, respiratory and nutritional support teams). RESULTS: 506 admissions in 390 patients with a mean age of 66.5 (18) years, 53% being women. The Charlson index was 2 (2.2) and the Barthel index 63.5 (40,4). Average stay was 7.9 (8.2) days. The main reasons for admission were the infections and domiciliary intravenous antibiotic therapy in 153 (30.5%) cases, followed by patients with chronic obstructive pulmonary disease or cardiac failure in 107 (21%) cases, and home enteral and parenteral nutrition in 102 (20%) cases. Two hundred (39.5) patients were subsequently controlled by their primary care team after discharger, 241 (47.5) patients were followed-up in the hospital consultations, and 45 (9%) of the patients had to return directly to the hospital. CONCLUSIONS: The creation of a HaHP, for medical diseases, in internal medicine department that is highly coordinated with medical services of the hospital, especially with emergency, respiratory, and nutritional support teams, and with the primary health care, facilitates specialized home care of medical diseases and improves coordination with the primary health care. PMID- 18381003 TI - [Bortezomib in relapsed or refractory multiple myeloma: results in a cohort of 39 patients]. AB - INTRODUCTION: Bortezomib has presently become a significant rescue treatment in multiple myeloma (MM) due to its observed effectiveness and safety in multicenter trials. We have aimed to verify both aspects in a setting of non-selected patients. PATIENTS AND METHODS: This is an observational, prospective study of the cohort of relapsed or refractory MM patients treated with bortezomib in our Department. The variables analyzed were response, its duration, time to the treatment failure (TTF), overall survival (OS), response related conditions and toxicity. Statistical methods used were Fisher's exact test, log rank-test and Kaplan-Meier survival tables. RESULTS: A total of 39 patients, 25 relapsed and 14 refractory to chemotherapy, started the treatment. The mean number of previous treatment was 2.3 and they received an average of 5.8 cycles of bortezomib. Complete response was achieved in 14 patients (36%), partial response in 12 (31%) and minor or no response in 13 ones (33%). Median duration of response was 8 months, median TTF was 10 months and median OS, from the onset of bortezomib was 16.5 months, with a median observation of live patients of 12.5 months. The response was more frequent in males (p = 0.019) and in patients with one previous treatment (p = 0.15). There were no significant differences regarding to TTF when we considered the cause of treatment (relapse or no response to chemotherapy) nor in the number of previous treatment regimes. The most frequent adverse events were reversible thrombocytopenia (31%), polyneuropathy (28%) and asthenia anorexia (23%). CONCLUSIONS: In our cohort of non-selected, relapsed or refractory MM patients, the observations found in the multicenter randomized trials results regarding response rate and duration, TTF OS and safety of bortezomib therapy were verified. PMID- 18381004 TI - [Creutzfeldt-Jakob disease: two new cases in Talavera de la Reina]. AB - Creutzfeldt-Jakob disease is a prion spongiform encephalopathy that causes a rapidly progressive dementia whose neuropathological features are neuronal loss, proliferation of glial cells, and the presence of small vacuoles, which produces a spongiform appearance. Two cases detected in the year 2005 in the <> Hospital are presented. The first one is the case of a man who presented with intense malaise and weight loss, developing dementia in 4 months. The second one was a woman who presented with diplopia, headache and gait disturbances who developed severe neurological impairment with dementia in three months. In both cases the 14.3.3 protein in the cerebrospinal fluid was positive and the EEG showed a characteristic pattern. The genetic studies were negative and both were homozygous for methionine at codon 129. Given that the incidence of this disease in Castilla la Mancha is 0.6 cases/100000 inhabitants/year, the detection of 2 cases in less than a year in Talavera de la Reina (whose population is 150,000) surprised us, leading us to intensify surveillance. PMID- 18381006 TI - [Exanthema after eating in the country]. PMID- 18381005 TI - [Bowel obstruction in patients under treatment with acenocumarol]. PMID- 18381007 TI - [Purpuric lesions and facial edema in adolescents after intense sun exposure]. PMID- 18381008 TI - [71-year old woman with nasal swelling]. PMID- 18381009 TI - [Neuro-Behcet and neurotoxicity due to cyclosporine]. PMID- 18381010 TI - [Mentonian hypoesthesis in a patient who smokes]. PMID- 18381011 TI - [Thomas Mann sign: deep venous thrombosis due to arteriosclerotic plaque rupture]. PMID- 18381013 TI - [Spontaneous chylothorax after minimum exercise in middle-aged woman. A disease to be recognized with good prognosis]. PMID- 18381014 TI - [Von Willebrand factor, endothelial lesion, and ischemic heart disease]. AB - OBJECTIVE: To analyze if the levels of von Willebrand factor (FvW) are higher in patients with ischemic heart disease than in healthy subjects and evaluate the relationship of these levels with clinical recurrence and coronary interventionism. DESIGN: Observational prospective study. PATIENTS: We analyzed the levels of FvW in 75 patients with ischemic heart disease who underwent coronary interventionism (Group I) and compared them with those of 30 healthy subjects with no cardiovascular risk factors and who, theoretically, had no coronary injuries (Group II). MAIN VARIABLES: Levels of FvW before coronary interventionism (sample 0), 24 hours after (sample 1), and at three months of out patient follow-up (sample 2). A single measurement was made of the FvW levels in Group II. RESULTS: Subjects with ischemic heart disease had higher levels of FvW than healthy subjects (162+/-74% versus 95+/-33%; p=0.0001). FvW levels were significantly increased after coronary interventionism (162.4+/-74.9% in sample 0 versus 213+/-90% in sample 1; p=0.0001). Patients with clinical symptoms at three months have no significant difference regarding those with no symptoms in the FvW levels (125+/-63% versus 133+/-60%; p=0.57). CONCLUSIONS: FvW levels reflect an endothelial alteration in patients with ischemic heart disease. The increase of the levels after coronary interventionism could be due to the endothelial aggression itself of the intervention. It was not possible to demonstrate higher levels of FvW in patients with symptoms in the three month follow-up. PMID- 18381015 TI - [Female gender is an independent predictor of in-hospital mortality in patients with ST segment elevation acute myocardial infarction treated with primary angioplasty]. AB - OBJECTIVE: The aim of this study was to determine the influence of gender on in hospital outcome in patients with ST-segment elevation myocardial infarction (STEMI) who underwent primary angioplasty (PA). DESIGN AND SCOPE: Prospective study of a cohort of patients consecutively admitted to the Coronary Unit of a tertiary hospital in the period of January to October 2004 with the diagnoses of IAMEST and treated with PA. PATIENTS: Consecutive sample of 86 patients with this diagnosis divided into two groups based on sex: 52 men and 34 women. MAIN VARIABLES OF INTEREST: In both groups, we analyzed the baseline clinical demographic characteristics, extension of the coronary disease (ECD), success of the PA, appearance of heart failure (HF) and in-hospital mortality in the first 28 days after admission. We analyzed predictors of mortality in a multivariate model. RESULTS: The women were older (70+/-8 versus 65+/-11; p=0.02) and had greater prevalence of diabetes (37% versus 18%; p=0.002) and hypertension (58% versus 37%; p<0.001) than the men while the men had greater frequency of smoking (34% versus 22%; p=0.001). There were no differences in the presence of hyperlipidemia, ECD or the success of PA. Women had a higher incidence of HF on admission (22% versus 12%; p=0.01) and in-hospital mortality (17% versus 8%; p=0.002). In the multivariate analyses, female sex and HF on admission continued to be predictors of in-hospital mortality. CONCLUSIONS: In our study, female gender was an independent predictor of in-hospital mortality in patients with IAMEST treated with PA. PMID- 18381016 TI - Comparison of bronchoscopic bronchoalveolar lavage vs blind lavage with a modified nasogastric tube in the etiologic diagnosis of ventilator-associated pneumonia. AB - OBJECTIVE: Our objective was to compare the results of a blind lavage vs a bronchoscopic-guided bronchoalveolar lavage for the etiologic diagnosis of ventilator-associated pneumonia (VAP). DESIGN: Prospective study in consecutive patients with high probability of VAP. Every patient underwent both procedures, in a formally randomized fashion. The interpretation of quantitative cultures was done in a blind fashion. SETTING: Single center study, with a 20 bed medical and surgical Intensive Care Unit of the University Hospital in Monterrey, Mexico. PATIENTS: Twenty-five patients with high probability of VAP. INTERVENTIONS: Every patient underwent blind bronchoalveolar lavage with a modified nasogastric tube, and a bronchoscopic-guided bronchoalveolar lavage. RESULTS: Twenty-one patients underwent both procedures. Four patients were excluded due to contamination of the cultures. The quantitative cultures were compared in a paired fashion. Only two patients had discordant cultures. The correlation coefficient between the number of colonies was very high, r=0.90 (95% confidence interval [CI], 0.77 0.96; p=0.0001). CONCLUSIONS: The blind bronchoalveolar lavage with a modified nasogastric tube is a valuable tool for the identification of etiologic agent in VAP, particularly when trained bronchoscopists or the necessary resources for bronchoscopic-guided bronchoalveolar lavage are not readily available. PMID- 18381017 TI - [Treatment recommendations at the end of the life of the critical patient]. AB - Admission of a patient in the Intensive Care Unit (ICU) is justified when the critical situation can be reverted or relieved. In spite of this, there is high mortality in the ICU in regard to other hospital departments. End-of-life treatment of critical patients and attention to the needs of their relatives is far from being adequate for several reasons: society denies or hides the death, it is very difficult to predict it accurately, treatment is frequently fragmented between different specialists and there is insufficient palliative medicine training, including communication skills. There are frequent conflicts related to the decisions made regarding the critical patients who are in the end of their life, above all, with the limitation of life-sustaining treatments. Most are conflicts of values between the different parties involved: the patient, his relatives and/or representatives, health professionals, and the institution. The SEMICYUC Working Group of Bioethics elaborates these Recommendations of treatment at the end of the life of the critical patient in order to contribute to the improvement of our daily practice in such a difficult field. After analyzing the role of the agents involved in decision making (patient, familiar, professional, and health care institutions), of the ethical and legal foundations of withholding and withdrawal of treatments, guidelines regarding sedation in the end of the life and withdrawal of mechanical ventilation are recommended. The role of advance directives in intensive medicine is clarified and a written form that reflects the decisions made is proposed. PMID- 18381018 TI - Significance of venous oximetry in the critically ill. AB - Critically ill patients are threatened or affected by multi-organ failure (MOF). Tissue hypoxia is one of the most important co-factors of MOF. Venous oximetry allows the critical estimation of the global oxygen (O(2)) supply-demand ratio and can be gained from mixed (S(v)O(2)) and central venous blood (S(cv)O(2)). Cellular requirements dominate the feedback hierarchy of the O(2)-metabolism. This review describes the history and validity of haemodynamic monitoring, illustrates the physiological background and clinical application of venous oximetry and presents carbon dioxide analysis as evidence of the usefulness of a multi-modal approach in cardio-respiratory monitoring. Variation of cardiac output, optimisation of arterial O(2)-saturation and adaptation of O(2) extraction are shown to be the relevant levels of pathophysiological adaptation as well as therapeutic intervention. We portray the functional equivalence of S(cv)O(2) and S(v)O(2) and analyse their diagnostic, therapeutic and prognostic significance, providing the basis for the efficacy of venous oximetry as an important marker of critical illness. Finally, having drawn an outline of current developments for the better understanding of the oxidative balance of individual organs, we stress the importance of a synoptic O(2)-monitoring strategy as well as the need to use its beneficial, yet unfulfilled, clinical potential. PMID- 18381019 TI - [A near-miss event. A new approach in the study of adverse events]. AB - A near-miss event is defined as an event that could have resulted in an injury, fatality, or property damage if it had not been prevented. Analysis of near-miss events could be an efficient method in the study of adverse events. Reporting of near-misses has many benefits in the study of adverse events since near-misses occur more frequently than adverse events. In addition, as they have no consequences, fear that the professionals involved would have to report them is less. However, up to now, this method has been slow to develop. We present two clinical cases that help to understand the usefulness of the near-miss reporting system. PMID- 18381020 TI - [MELAS: diagnostic keys and treatment in the Intensive Care Unit]. AB - Cerebrovascular accident in young adults is not rare and the most common causes are: cardiac embolism, arterial dissection and migraine. The unknown etiology, in spite of extensive studies, is described in the literature in about one third of patients under 45 years of age. The mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS) may occur with symptoms that simulate a stroke. The <> episodes generally affect the posterior part of the brain and the basic insult mechanisms may be metabolic more than ischemic. We describe the case of an 18-year old patient who was admitted to the Intensive Care Unit due to a sudden picture of blindness. The ophthalmologic study was normal and the neuroimaging studies showed infarction in both occipital lobules. We explain the different clinical features, diagnostic, therapeutic and prognostic methods of the disease. PMID- 18381021 TI - [Severe intoxication with verapamil]. PMID- 18381022 TI - [Loffler syndrome in post-operative of heart surgery]. PMID- 18381023 TI - [Comments on the recommendations for antibiotic treatment of gram positive coccal infections in the critical patient]. PMID- 18381025 TI - [Images in intensive medicine]. PMID- 18381026 TI - [Role of pulse pressure, systolic blood pressure, and diastolic blood pressure in the prediction of cardiovascular risk. Cohort study]. AB - BACKGROUND AND OBJECTIVE: To analize the role of pulse pressure (PP), systolic (SBP) and diastolic blood pressure (DBP), in the prediction of cardiovascular risk. PATIENTS AND METHOD: A prospective cohort study carried out in 2 primary care center, including 932 patients aged between 35-84 years old, without cardiovascular events, selected by simple random sampling, and with an 8 year follow-up. PP, SBP, and DBP were categorized in tertiles, comparing the upper with the 2 lowers. First cardiovascular event, whether fatal or not, such as coronary heart disease, cerebrovascular disease, and peripheral arterial disease was recorded as a composite variable. Results were studied according to Cox models, adjusting for age, sex, smoking, total cholesterol, high density lipoprotein cholesterol, and diabetes mellitus. We studied the correlation between PP with SBP, DBP, and cardiovascular risk factors. RESULTS: We registered 85 cardiovascular events: 43 cases of coronary heart disease, 27 cerebrovascular disease, and 17 peripheral arterial disease. The adjusted hazard ratios for composite variable were: upper PP tertile (>/= 59 mmHg) = 1.3 (95% confidence interval [CI], 0.8-2.1); upper SBP tertile (>/= 140 mmHg) = 1.5 (95% CI, 1.0 2.5); upper DBP tertile (>/= 84 mmHg) = 1.1 (95% CI, 0.7-1.8). Results were similar for specific cardiovascular events. PP was correlated with SBP (r = 0.825; p < 0.001), age (r = 0.422; p < 0.001), diabetes mellitus (r = 0.242; p < 0.001), and smoking (r = -0.158; p = 0.01), with adjusted hazard ratio for these variables of 1.0 (95% CI, 0.6-1.9). CONCLUSIONS: PP is an arterial pressure component very correlated with SBP and other factors, but is not a better cardiovascular risk predictor than SBP. PMID- 18381027 TI - [Antibiotic treatment onset delay and clinical evolution in admitted patients with community-acquired pneumonia]. AB - BACKGROUND AND OBJECTIVE: The delay in the initiation of antibiotic treatment in patients with community-acquired pneumonia (CAP) could influence their prognostic. The aim of our study was to evaluate the influence of a delay in antibiotic treatment on mortality, occurrence of complications, and length of stay in the hospital, in relation to the prior duration of the fever. PATIENTS AND METHOD: This is a cohort study which includes all patients over 18 years of age admitted with CAP over a 4 year period. The response variables analysed were in-hospital mortality, average stay in days and occurrence of complications during evolution. The relationship between the delay in antibiotic treatment and the outcome variables was estimated by logistic regression models, adjusting the estimations by PSI (Pneumonia Severity Index) and APACHE II (Acute Physiology and Chronic Health Evaluation). RESULTS: Six hundred fifty-nine patients were included (average age of 71 years, 71% males). Seventy-six (11.6%) patients died, 12.1% of those who arrived after 48 hours (55.4% of patients) and 10.9% of those who came early. The delay in antibiotic treatment was associated with a higher mortality (odds ratio [OR] = 3.05; 95% confidence interval [CI], 0.86-10.9; threshold of 8 h), prolonged stay (OR = 2.38; CI 95%, 1.11-5.14; threshold of 5 h) and a higher rate of complications (OR = 2.24; CI 95%, 0.90-5.53; threshold of 3 h). CONCLUSIONS: Our results suggest the importance of the early administration of antibiotics in patients with CAP. There is also an increased risk for each of these outcomes in those patients with a history of fever of more than 2 days before arriving in hospital. PMID- 18381029 TI - [Pulse pressure and cardiovascular risk: from epidemiology to pathophysiology]. PMID- 18381028 TI - [Efficacy of gabapentin in the treatment of carpal tunnel syndrome]. AB - BACKGROUND AND OBJECTIVE: To evaluate the analgesic efficacy and safety of gabapentin in the treatment of carpal tunnel syndrome (CTS), as well as the electromyographic (EMG) evolution after 6 months. PATIENTS AND METHOD: A prospective study with a 6-month follow-up of patients with EMG diagnosis of primary CTS starting treatment with 1.800 mg/day of gabapentin. At baseline visit and after 6 months of treatment a complete clinical evaluation and an EMG study were performed. Adverse effects of gabapentin were also registered. RESULTS: Twenty-five patients were included, mean age (standard deviation) 58.88 (7.69) years. After 6 months of treatment, a statistically significant reduction of pain (p = 0.001) and improvement of severity of symptoms (p = 0.008) were observed, although functional capacity did not change. EMG was performed in 19 patients at 6 months. Compared to baseline EMG: 52.6% patients showed no changes in EMG findings, while 5.3% patients showed improvement and in 26.3% the EMG was normal. Progression was only seen in 15.8% of patients after 6 months of treatment. In 28% of the patients gabapentin was stopped because of side effects. CONCLUSIONS: In our series, gabapentin was effective in the reduction of pain and improvement of the severity of the symptoms. Results of EMG after 6 months of treatment showed no changes, with improvement and/or remission in 84.2% of the cases. The drug was safe and well tolerated. PMID- 18381030 TI - [Critical evaluation of clinical practice guidelines]. PMID- 18381031 TI - [Current strategies in the treatment of renal-cell cancer: targeted therapies]. AB - Renal-cell carcinoma represents 95% of all renal tumours. The Von Hippel-Lindau (VHL) tumor-suppressor gene is mutated or silenced in most clear cell renal carcinomas. pVHL loss results in the stabilization of the heterodimeric transcription factor hypoxia-inducible factor (HIF) and enhanced transactivation of HIF target genes. HIF itself has been difficult to inhibit with drug-like molecules although a number of agents that indirectly inhibit HIF, including mTOR (mammalian target of rapamycin) inhibitors, have been identified. Moreover, a number of drugs have been developed that target HIF-responsive gene products, such as vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), implicated in tumor angiogenesis. Many of these targeted therapies, especially sunitinib, have demonstrated significant activity in kidney cancer clinical trials and represent a substantive advance in the treatment of this disease. PMID- 18381032 TI - [Medical personnel's careers. A useful incentive system as long as we adhere to its application principles]. PMID- 18381033 TI - [Leukocytoclastic vasculitis and acenocoumarol]. PMID- 18381034 TI - [Hyperhomocysteinema, B12 deficit and thromboembolic venous disease]. PMID- 18381035 TI - [Coronary spasm and syncopal polimorphic ventricular tachycardia]. PMID- 18381036 TI - Multidrug-resistant Acinetobacter baumannii: eyes wide shut? PMID- 18381037 TI - [Tuberculosis in the Bajo Deba area (Guipuzcoa, Spain) from 1995 to 2006]. AB - BACKGROUND: Within the Multicenter Project on Tuberculosis Research performed in Spain in 1996-1997, the Bajo Deba Area reported the highest incidence of tuberculosis in the Basque Country. We analyzed the clinical and epidemiological characteristics of the tuberculosis population diagnosed in our area during the period of 1995 to 2006. METHODS: Ambispective, observational study. RESULTS: A total of 584 patients were diagnosed with tuberculosis. The disease affected the respiratory tract in 509 cases and other sites in 75 cases. The mean annual incidence rate of tuberculosis was 64.5 cases per 100 000 inhabitants (91.6 in 1995-1998; 34.9 in 2003-2006). The mean annual incidence rate of smear-positive patients was 20.7 cases per 100 000 inhabitants (33.8 in 1995-1998; 12.9 in 2003 2006). The 15 to 24-year-old group was the most highly affected during the period of 1995 to 1998 (mean annual incidence rate 199.4 cases per 100 000); in contrast, the > 75-year-old group was the most highly affected during the period of 2003 to 2006 (121.1 cases per 100 000 inhabitants). Fifty-three patients were co-infected by HIV (9%) (yearly mean of 11.6% in 1995-1998 and 7% in 2003-2006). Lowenstein culture was positive in 431 cases (73.8%). Resistance to isoniazid was detected in 1.4% out of a total of 287 strains tested, and multidrug resistance was not observed. Nine patients were immigrants (1.5%). Treatment completion was greater in our area (505 patients, 86.4%), as compared to that recorded in the Guipuzcoa province during the same period (1956 of 2525 patients, 77.5%) (P < .01). CONCLUSIONS: The Bajo Deba Area presented a high incidence of tuberculosis in the 1995 to 2006 period. Epidemiological trends showed a progressive decrease in the number of tuberculosis patients, with a shift from younger to older persons as the most highly affected age group. The impact of drug resistance and immigration was negligible on tuberculosis rates. The percentage of microbiologically confirmed cases was high. Treatment completion was satisfactory. PMID- 18381038 TI - [Vaccination coverage against Streptococcus pneumoniae in splenectomized patients in a fourth-level hospital (1999-2004)]. AB - INTRODUCTION: Patients in whom the spleen has been removed have a higher risk of acquiring infection by capsulated microorganisms, particularly Streptococcus pneumoniae, Haemophilus influenzae type b, and Neisseria meningitidis. AIMS: Determine the vaccination coverage against S. pneumoniae of splenectomized patients in Hospital Clinico San Carlos (HCSC, Madrid) and to identify variables related to the level of vaccination coverage reached. METHODS: From January 1, 1999 to December 31, 2004, a retrospective cohort study of all splenectomized patients was carried out in HCSC. Patients were identified from the Medical Records Department and the vaccination registry of the Preventive Medicine Service. RESULTS: During the study period, 248 splenectomies were carried out. The median age was 61.0 years (interquartile range, 39.0-71.0). The most frequent reason for splenectomy was malignant gastrointestinal neoplasm (34.7%). Overall vaccination coverage against S. pneumoniae for the period studied was 48.4%. A significant increase in vaccination coverage per year was observed (P < .001). Multivariate analysis identified the following variables associated with a greater risk of not being vaccinated: male sex, patients proceeding from surgical services, and patients undergoing splenectomy during 1999 to 2001. CONCLUSIONS: Vaccination coverage has increased every year since implementation of the protocol. Nevertheless, it necessary to extend the vaccination program within daily clinical practice. PMID- 18381039 TI - [Prevalence of infection by carbapenem-resistant Acinetobacter baumannii in Spain (1999-2005)]. AB - INTRODUCTION: This study determines the prevalence of infections by carbapenem resistant Acinetobacter baumannii (CRAB) in Spain, and identifies trends over time, geographical variations, and factors associated with resistance. METHODS: Yearly prevalence surveys (EPINE, Estudio de Prevalencia de las Infecciones Nosocomiales en Espana) during 1999 to 2005 were analyzed, including data on the site of infection, culture, and antimicrobial susceptibility, and characteristics of the patients and hospitals. RESULTS: On average, 246 hospitals/year participated in the survey. A total of 1168 A. baumannii isolates were identified, yielding an infection prevalence rate of 3/1,000 hospitalized patients. Fourteen percent of isolates were related to community-onset infections. The most frequent sites of infection were the respiratory tract (42.2%), surgical wound (15.1%), urinary tract (12.9%), and skin (11.7%). Rate of carbapenem resistance was 34.5% (95% CI, 31.8-37.3), and was even higher among ICU patients (43.8%; 95% CI, 38.9%-48.7%). There were considerable differences between Spanish regions, with the highest rates of resistance in central regions. Higher resistance rates were observed in respiratory tract infections (43%) and catheter-related bacteremia (47.6%) than among other sites of infection (P =.003). Main factors associated with CRAB identified by multivariate analysis were surgical or tracheostomy procedures, use of invasive devices such as urinary, nasogastric, or central venous catheters, and mechanical ventilation, as well as male gender and pressure sores. CONCLUSIONS: The prevalence of CRAB in Spain is very high. Differences in resistance rates have been observed according to geographic region and area of hospitalization. Several invasive procedures, as well as male gender and pressure sores, are associated with higher rates of carbapenem resistance. PMID- 18381040 TI - [Cutaneous tuberculosis: a 15-year descriptive study]. AB - INTRODUCTION: Cutaneous tuberculosis has experienced a resurgence following a period of decline. The aim of this study was to determine the clinical and epidemiological characteristics of cutaneous tuberculosis in the Ferrol Healthcare Area (Spain). METHODS: Between 1991-2005, 1139 new cases of tuberculosis were diagnosed in Ferrol and submitted to a descriptive analysis. Cutaneous involvement was investigated in all cases. RESULTS: Among 1139 patients, 55 cases of cutaneous tuberculosis were diagnosed (4.8%). The condition was more frequent in women (70.9%), average age was 44.1 +/- 23.3 years, and 56.4% of patients were treated with 6HR2Z-E. Among the 55 cases, 26 (2.3%) were true cutaneous tuberculosis and 29 (2.5%) were tuberculids. In the 26 cases of true tuberculosis, the most frequent form was scrofuloderma (32.7%), followed of lupus vulgaris (7.2%), tuberculous gumma (3.6%), and tuberculosis verrucosa cutis (3.6%). The most frequent locations were the neck, face and trunk, and in 76.9% other sites were involved (lymph nodes 14, bone 6, lung 4, and intestine 1). In the 29 cases with tuberculids, erythema nodosum was the most frequent form (49.1%) followed by erythema induratum of Bazin (3.6%). The lower extremities were affected in all cases and there was simultaneous involvement of other site in 51.9%. In patients with true cutaneous tuberculosis, the diagnostic yield was greater (necrotizing granulomas in 70.6% of biopsies and positive Lowenstein culture for Mycobacterium tuberculosis in 77.8%) and average age was higher than in patients with tuberculids (P < .05). CONCLUSION: Cutaneous tuberculosis is uncommon, preferentially affects women, and is usually associated with tuberculous disease in other locations, particularly in the case of scrofuloderma. PMID- 18381041 TI - [Bone mineral loss in patients with human immunodeficiency virus infection]. AB - Life expectancy and quality of life among patients infected with human immunodeficiency virus (HIV) has dramatically improved with the advent of highly active antiretroviral therapy. Nonetheless, with the transformation of HIV infection into a chronic disease, a series of long-term consequences have been observed, among them osteoporosis. The prevalence of this condition is increasing among individuals with HIV infection. Although an epidemic of fragility fractures has not been described in these patients, an increase in the incidence of such fractures is anticipated as the HIV population ages, and this will undoubtedly have an impact on their quality of life. The numerous factors that favor the development of osteoporosis in these patients are related to the infection, itself, and the treatment received. PMID- 18381042 TI - [Epidemiological surveillance cultures in antimicrobial-resistant bacteria causing nosocomial infection]. AB - Implementation of surveillance culture programs and molecular typing are important contributions of Clinical Microbiology to the control of nosocomial infections. This document provides information on collection, transport, preservation, and processing of samples for surveillance culture, as well as the criteria for interpreting and reporting the results of relevant etiologic agents in nosocomial infection. This includes methicillin-resistant Staphylococcus aureus (MRSA), glycopeptide-resistant Enterococcus spp., enterobacteria producing extended-spectrum beta -lactamases (ESBLs), multiresistant Acinetobacter baumannii and carbapenem-resistant Pseudomonas aeruginosa. Details on the available methods for rapid diagnosis are also presented. The information in this document attempts to provide a general approach to the problem and may be considered a starting point for laboratories that are developing their own guidelines, according to needs defined by the multidisciplinary nosocomial infection control team. PMID- 18381043 TI - [Recommendations for antibiotic monitoring in ICU patients]. AB - Monitoring plasma concentrations of antimicrobial agents used to treat infection in critically ill patients is one of the recommended strategies for improving clinical outcome. Drug monitoring has a double AIM: to limit adverse events and to increase the effectiveness of the drugs. In clinical practice, however, this approach is mainly limited to monitoring plasma concentrations of vancomycin and aminoglycosides, although future extension to other antimicrobial agents would be desirable. Application of this technique varies considerably between hospitals, and this makes interpretation and comparison of the results obtained difficult. For this reason, representatives of various scientific societies related to the pharmacokinetic area have developed a series of recommendations for monitoring plasma concentrations of antimicrobials using vancomycin and several aminoglycosides as the reference. The recommendations are based on 14 questions encompassing all steps of the process: indication for the test, blood sampling (timing of blood collection, blood volume, tubes), transport to the laboratory, techniques applied, normal values, dose adjustment, and reporting the RESULTS: The purpose of these guidelines is to develop a process of monitoring plasma antimicrobial concentrations that is as homogeneous as possible to facilitate the design of multicenter studies, as well as the interpretation and comparison of results. PMID- 18381044 TI - [Infection of cerebrospinal fluid shunt systems]. AB - Cerebrospinal fluid (CSF) shunt infection is a cause of considerable morbidity and mortality. Shunt infection is produced mainly during surgery and by surgical wound infection. Staphylococcus spp. (> 50% methicillin-resistant) are the most common causative bacteria, although gram-negative bacilli (10%-25%) and Propionibacterium acnes are becoming increasingly implicated. Shunt malfunction syndrome and fever are the most frequent clinical manifestations, whereas signs of meningeal irritation are uncommon. Other clinical manifestations depend on the location of the distal catheter. CSF should be obtained by puncture of the shunt reservoir or the distal catheter and processed for biochemical analyses, cell count, Gram stain, and aerobic and anaerobic cultures (lengthy incubation). Because of biofilm formation and to avoid recurrences, the recommended treatment is intravenous antibiotics plus removal of all components of the infected shunt, followed by placement of an external drainage catheter and a new shunt. Prophylaxis is important and can include antimicrobial prophylaxis and/or antibiotic-impregnated catheters. PMID- 18381045 TI - [Ulcerous lesion on the right arm of an HIV-positive patient]. PMID- 18381046 TI - [Panophthalmitis following topical cocaine use in an HIV+ patient]. PMID- 18381047 TI - [Acute Streptococcus suis meningitis in a woman working in a meat market]. PMID- 18381048 TI - [Herpes esophagitis in an immunocompetent patient]. PMID- 18381049 TI - [Epidemiological surveillance of nosocomial infection by a multidisciplinary infection control team]. PMID- 18381050 TI - [Visceral leishmaniasis in recently diagnosed HIV: immune reconstitution syndrome?]. PMID- 18381051 TI - [Sex differences in Alzheimer's disease]. AB - The prevalence of Alzheimer disease is higher in women than in men. In the age group 65-69 years 0.7% of women and 0.6% of men suffer from the disease with increasing frequencies of 14.2% and 8.8% in individuals aged 85-89 years. The incidence is also higher in demented women. In Austria 74.1% of Alzheimer patients older than 60 years are women. Several studies report more pronounced language, mnestic, semantic and orientation deficits in women, but methodological shortcomings might be responsible for this finding. The validity of results reporting a more rapid cognitive decline in women can also be questioned. Women have a broader spectrum of dementiarelated behavioural symptoms with a predominance of depression, while aggression is more frequent in men than in women. Biological explanations for gender-specific differences in the phenotype of Alzheimer s disease include different brain morphology and function with higher susceptibility for pathological lesions in women and greater cognitive reserve in men. Sex differences were also reported for expression of antioxidative enzymes and post-menopausal hormonal changes. Interactions between gender nd response to treatment, if any, are subtle and have large intra individual variability. In Austria, two thirds of patients receiving attendance allowance are women. Care takes place in 80% by the families and is provided by women in 78%. The rate of female care-givers in partly institutionalized care units in 91% in nursing homes it is 84%. PMID- 18381052 TI - [Does antidepressant therapy increase suicide risk in children ad adolescents? - A comment]. AB - Recent data indicate increasing suicide rates for children and adolescents in the United States, Great Britain and the Netherlands. These facts call for a critical discussion of prescription rates of antidepressants for pediatric use. Obviously the U.S. and European regulators issued public warnings about a possible association between antidepressants and suicidal thinking and behaviour have discouraged physicians to use antidepressants in this age group. Untreated depression means impairment of psychosocial development of children and their families. Alarmingly there is no evidence of a significant increase in the use of treatment alternatives as for example psychotherapy. High relapse rate with early onset of depressions are common and therefore children with depression need all our attention and best treatment available. Current Austrian suicidal statistics do not yet demonstrate increasing suicidal rates in adolescents as seen in the U.S., the U.K. and the Netherlands. Thus all our alertness is needed to avoid any analogical progress in suicide rates and despite regulatory warnings and certainly after a careful risk-benefit analyses physicians have to consider pharmacological treatment options in depressive children and adolescents. PMID- 18381053 TI - [Insights in the central serotonergic function in patients with affective disorders]. AB - OBJECTIVE: The simple insight in the central serotonergic activity would allow a more specific pharmacotherapy of disorders with serotonergic deficits and could thus serve as a predictive parameter. An antidepressive treatment could be chosen due to the verification or falsification of such a deficit with a serotonergic or noradrenergic substance. In search of predictive parameters we have investigated the loudness dependence of auditory evoked potentials (LDAEP), a parameter that has already been discussed concerning this matter, and serotonin levels in the human blood serum regarding their correlation. METHODS: Eight outpatients of a lithium clinics were investigated neurophysiologically after blood samples had been taken. RESULTS: A significant negative correlation between LDAEP and serotonin level in the serum was found, a low serotonin level correlates with a high LDAEP. CONCLUSIONS: The result of this pilot study justifies further investigations with larger sample sizes. In case of further and stable evidence the use of serotonin levels in the serum as a predicitve parameter is conceivable. PMID- 18381054 TI - [Origin of patients admitted involuntarily from an urban catchment area]. AB - OBJECTIVE: This investigation highlights some aspects of migration of patients between catchment areas. METHODS: From January till June 2003 all committed patients admitted from Linz, capital of the Austrian province Upper Austria, were investigated for their origin. RESULTS: Out of a total of 214 patients 111 (52%) were not native to Linz. Most stemmed from other counties of Upper Austria (55%), 16 (14%) were from other Austrian provinces and the remainder 32 (29%) were from foreign countries. In patients from other counties of Upper Austria 59% were already psychiatric ill before they had moved to Linz. Compared with patients native to Linz they displayed a greater number of psychiatric admissions, a greater number of cumulative hospital days and more often needed some measures of supported housing. More than a quarter of them had moved to Linz in the last 5 years. CONCLUSIONS: Results show that also in a well-established system of community psychiatric care a considerable number of patients is moving towards urban areas. PMID- 18381055 TI - [Alzheimer's disease: knowledge and attitudes in a representative survey]. AB - Dementia is a prevalent syndrome in ageing societies and therefore of significant medical and social importance for the general population. We have studied knowledge and attitudes towards Alzheimer's dementia (AD) of 1245 epidemiologically representative individuals between 14 and 99 years. Only 13% mentioned memory disturbances, e.g. forgetfulness, as hallmarks of AD; 54% knew that age was a major risk factor; 47% felt that "brain-jogging" was therapeutically useful. In case of developing AD, more than 70% wished to be informed together with a close relative or friend; 7% felt that nobody else should know about their problem; and many more than 50% expected information on treatment, course, symptoms and causes. These results demonstrate, that there is a remarkable lack of relevant information on AD in the general population, and that most individuals wished to be informed about a potential diagnosis of AD together with their family and friends. PMID- 18381056 TI - [Neuropsychiatric symptoms in Sotos syndrome. Case report and review of the literature]. AB - Sotos syndrome, or cerebral gigantism, is a rare genetic syndrome characterized by excessive growth during childhood, macrocephaly, distinctive facial gestalt and learning difficulties. It is caused by mutations or deletions of the NSD-1 gene. Most cases are sporadic. Apart from a number of physical abnormalities that are commonly present, a high prevalence of cognitive, emotional and behavioural problems in children with Sotos syndrome can be assumed. However, there has been almost no literature about psychiatric symptoms in adults with Sotos syndrome so far; one case of psychosis was reported. In the present case, the authors present psychopathological features of an adult patient with Sotos syndrom who developed among other things - psychotic symptoms. PMID- 18381057 TI - [Women seek for help - men die! Is depression really a female disease?]. AB - Prevalence rates of depression in females are about two to three times higher as compared to men. Biological evidence seems to support these data. Genderspecific suiciderates, closely linked to depressive illness, however raise doubts about the genderspecific epidemiological data of depression as males commit suicide three to ten times more often than females. The sociomedical concept of "male depression" delivers an explanation. A gender bias in diagnose as well as a reduced male helpseeking behavior and dysfuntional stress coping in males might be reasons for the reported low prevalence-rates of depression in males. Depression might hide behind addictive- and risk-behavior as well as irritability. As these symptoms differ from the classic depressive symptoms they might not be detected and identified as such. Validated rating scales which specifically assess male symptoms of depression, as well as peer-education programs for colleagues might change current depression rates in males. PMID- 18381058 TI - A prion disease of cervids: chronic wasting disease. AB - Chronic wasting disease (CWD) is a prion disease of deer, elk, and moose, initially recognized in Colorado mule deer. The discovery of CWD beyond the borders of Colorado and Wyoming, in Canada and as far east as New York, has led to its emergence as a prion disease of international importance. Epidemiological studies indicate that CWD is horizontally transmitted among free-ranging animals, potentially indirectly by prion-containing secreta or excreta contaminating the environment. Experimental CWD transmission attempts to other wild and domestic mammals and to transgenic mice expressing the prion protein of cattle, sheep, and humans have shed light on CWD species barriers. Transgenic mice expressing the cervid prion protein have proven useful for assessing the genetic influences of Prnp polymorphisms on CWD susceptibility. Accumulating evidence of CWD pathogenesis indicates that the misfolded prion protein or prion infectivity seems to be widely disseminated in many nonneural organs and in blood. This review highlights contemporary research findings in this prion disease of free ranging wildlife. PMID- 18381059 TI - Signaling by the human serotonin(1A) receptor is impaired in cellular model of Smith-Lemli-Opitz Syndrome. AB - The Smith-Lemli-Opitz Syndrome (SLOS) is a congenital and developmental malformation syndrome associated with defective cholesterol biosynthesis. SLOS is clinically diagnosed by reduced plasma levels of cholesterol along with elevated levels of 7-dehydrocholesterol (and its positional isomer 8-dehydrocholesterol) and the ratio of their concentrations to that of cholesterol. Since SLOS is associated with neurological deformities and malfunction, exploring the function of neuronal receptors and their interaction with membrane cholesterol under these conditions assumes significance. We have earlier shown the requirement of membrane cholesterol for the ligand binding function of an important neurotransmitter G-protein coupled receptor, the serotonin(1A) receptor. In the present work, we have generated a cellular model of SLOS using CHO cells stably expressing the human serotonin(1A) receptor. This was achieved by metabolically inhibiting the biosynthesis of cholesterol, utilizing a specific inhibitor (AY 9944) of the enzyme required in the final step of cholesterol biosynthesis. We utilized this cellular model to monitor the function of the human serotonin(1A) receptor under SLOS-like condition. Our results show that ligand binding activity, G-protein coupling and downstream signaling of serotonin(1A) receptors are impaired in SLOS-like condition, although the membrane receptor level does not exhibit any reduction. Importantly, metabolic replenishment of cholesterol using serum partially restored the ligand binding activity of the serotonin(1A) receptor. These results are potentially useful in developing strategies for the future treatment of the disease since intake of dietary cholesterol is the only feasible treatment for SLOS patients. PMID- 18381060 TI - Lipid organization in human and porcine stratum corneum differs widely, while lipid mixtures with porcine ceramides model human stratum corneum lipid organization very closely. AB - The conformational disordering and lateral packing of lipids in porcine and human isolated stratum corneum (SC) was compared using Fourier transform infrared spectroscopy (FTIR). It was shown that SC of both species differ markedly, porcine SC lipids being arranged predominantly in a hexagonal lattice while lipids in human SC are predominantly packed in the denser orthorhombic lattice. However, the lipid organization of equimolar ceramide:cholesterol:free fatty acid (CER:CHOL:FFA) mixtures prepared with isolated porcine CER or human CER is very similar, only the transition temperatures differed being slightly lower in mixtures with porcine CER. Therefore, the difference in lateral packing between human and porcine stratum corneum is not due to the difference in CER composition. Furthermore, it is possible to use more readily available porcine CER in model lipid mixtures to mimic lipid organization in human SC. As the equimolar porcine CER:CHOL:FFA mixtures closely mimic the lipid organization in human SC, both human SC and this mixture were selected to examine the effect of glycerol on the lipid phase behaviour. It was found that high concentrations of glycerol change the lamellar organization slightly, while domains with an orthorhombic lateral packing are still observed. PMID- 18381061 TI - Globus pallidus plays a critical role in neurotrophic factor induced functional improvements in hemiparkinsonian monkeys. AB - This study was designed to test the hypothesis that the integrity of the globus pallidus (GP) is critical for neurotrophic factor, such as glial-derived neurotrophic factor (GDNF), induced functional changes in rhesus macaques with MPTP-induced parkinsonism, because our previous studies demonstrated that the GP was one of the most affected areas as assessed by the levels of dopamine (DA) and its metabolites. A group of eight hemiparkinsonian monkeys with pallidal lesions, which positively responsed to intraventricular (ICV) injections of GDNF prior to the lesions, and a group of eight hemiparkinsonian monkeys without pallidal lesions, were treated with GDNF after a long washout period after the initial ICV infusions of GDNF. Significant behavioral improvements were only seen in the monkeys without pallidal lesions that received GDNF. Monkeys with pallidal lesions failed to exhibit any behavioral improvement even though they had elevated nigral DA levels. The results suggest that the GP is critical for neurotrophic factor induced functional changes in PD monkeys. PMID- 18381062 TI - Receptor expression is essential for proliferation induced by dimerized Jak kinases. AB - Two members of Jak kinases, Jak1 and Jak3, are associated with the cytoplasmic domains of the interleukin-2 (IL-2) receptor (IL-2R) beta chain (IL-2Rbeta) and the common cytokine receptor gamma chain (gammac), respectively, and accumulating evidence indicates their functional importance in IL-2 signaling. Here, I showed that coumermycin-induced chemical heterodimerization of Jak1 and Jak3 but not homodimerization of Jak1 or Jak3 induces cell proliferation of an IL-2R reconstituted cell line. In this regard, expression of IL-2Rbeta was essential for cell proliferation by chemical heterodimerization of Jak1 and Jak3, indicating that dimerized Jak1 and Jak3 induce heterodimerization of IL-2Rbeta and gammac, which may activate receptor-bound signaling molecules. Previous reports using chemical dimerization suggest that dimerization of Jak kinases is sufficient to induce cell proliferation. The present study indicates that re evaluation of this conclusion is necessary and that interpretation of functional analysis of signaling molecules using chemical dimerizers needs more careful assessment. PMID- 18381063 TI - The orphan nuclear receptor DAX-1 acts as a novel transcriptional corepressor of PPARgamma. AB - DAX-1 is an atypical nuclear receptor (NR) which functions primarily as a transcriptional corepressor of other NRs via heterodimerization. Peroxisome proliferator-activated receptor (PPAR) gamma is a ligand-dependent NR which performs a key function in adipogenesis. In this study, we evaluated a novel cross-talk mechanism between DAX-1 and PPARgamma. Transient transfection assays demonstrated that DAX-1 inhibits the transactivity of PPARgamma in a dose dependent manner. DAX-1 directly competed with the PPARgamma coactivator (PGC) 1alpha for binding to PPARgamma. Endogenous levels of DAX-1 were significantly lower in differentiated 3T3-L1 adipocytes as compared to preadipocytes. Using a retroviral expression system, we demonstrated that DAX-1 overexpression downregulates the expression of PPARgamma target genes, resulting in an attenuation of adipogenesis in 3T3-L1 cells. Our results suggest that DAX-1 acts as a corepressor of PPARgamma and performs a potential function in the regulation of PPARgamma-mediated cellular differentiation. PMID- 18381064 TI - Thermodynamic analysis reveals that GTP binding affects the interaction between the alpha- and gamma-subunits of translation initiation factor 2. AB - Eukaryotic and archaeal translation initiation factors 2, heterotrimers that consist of alpha-, beta-, and gamma-subunits, deliver methionylated initiator tRNA to a small ribosomal subunit in a manner that depends on GTP. To evaluate correlation of the function and association of the subunits, we used isothermal titration calorimetry to analyze the thermodynamics of the interactions between the alpha- and gamma-subunits in the presence or absence of a nonhydrolyzable GTP analog or GDP. The alpha-subunits bound to the gamma-subunit with large heat capacity change (DeltaC(p)) values. The DeltaH and DeltaC(p) values for the interaction between the alpha- and gamma-subunits varied in the presence of the GTP analog but not in the presence of GDP. These results suggest that the binding of both the alpha-subunit and GTP changes the conformation of the switch region of the gamma-subunit and increases the affinity of the gamma-subunit for tRNA. PMID- 18381065 TI - Identification of a novel 4-hydroxyphenylpyruvate dioxygenase from the soil metagenome. AB - 4-Hydroxyphenylpyruvate dioxygenase (HPPD) is a Fe(II)-dependent, non-heme oxygenase that converts 4-hydroxyphenylpyruvate to homogentisate. Essential cofactors, such as plastoquinone and tocopherol, are produced by HPPD-dependent anabolic pathways in plants. To isolate a novel hppd using culture-independent method, a cosmid metagenomic library was constructed from soil in Korea. Screening of Escherichia coli metagenomic libraries led to the identification of a positive clone, YS103B, producing dark brown pigment in Luria-Bertani medium supplemented with l-tyrosine. In vitro transposon mutagenesis of YS103B showed that the 1.3kb insert was sufficient to produce the hemolytic brown pigment. Sequence analysis of YS103B disclosed one open reading frame encoding a 41.4kDa protein with the well-conserved prokaryotic oxygenase motif of the HPPD family of enzymes. The HPPD-specific beta-triketone herbicide, sulcotrione, inhibited YS103B pigmentation. The recombinant protein expressed in E. coli generated homogentisic acid. Thus, we present the successful heterologous expression of a previously uncharacterized hppd gene from an uncultured soil bacterium. PMID- 18381066 TI - Chronic activation of AMP-activated protein kinase-alpha1 in liver leads to decreased adiposity in mice. AB - To assess the metabolic effects of chronic activation of AMP-activated protein kinase (AMPK) in liver, we generated a new transgenic (Tg) mouse model expressing constitutively active (CA)-AMPK-alpha1 in liver. In the short-term activation, the TgCA-AMPK-alpha1 mice exhibited minimal phenotype, but the Tg liver had elevated sterol regulatory element-binding protein (SREBP)-2 mRNA level and a parallel increase in transcripts of its target genes. UCP2 mRNA level was elevated. In the long-term activation, the TgCA-AMPK-alpha1 mice had markedly reduced white fat mass. The Tg liver had reduced mRNA expression of SREBP-1c and its target genes. Remarkably, the Tg mice were resistant to a high-fat diet induced obesity. These data suggest that short-term chronic activation of AMPK alpha1 in liver leads to compensatory increase in lipogenic gene expression due to increased SREBP-2 expression, and long-term chronic activation of AMPK-alpha1 decreases expression of SREBP-1c and its target genes, which results in reduced fat storage. PMID- 18381067 TI - Higher mortality in heterozygous neuropilin-1 mice after cardiac pressure overload. AB - We previously identified that neuropilin-1 (NP-1) was a co-receptor of vascular endothelial growth factor receptor 2 (VEGFR2) and confirmed that NP-1 knockout mice were embryonic lethal due to impairment of vascular development, while VEGF was reported to be involved in the progression of heart failure. However, it is unknown whether NP-1 has any influence on cardiac function, and it also remains poor understood concerning cardiac expression of NP-1 and its interaction with other VEGF receptors in the heart. Here, we first showed that NP-1 heterozygous mice had significantly higher mortality due to either acute or chronic heart failure in response to left ventricular pressure overload. We also observed that NP-1 mRNA and protein were expressed in both neonatal rat cardiomyocytes and adult murine heart. Furthermore, we found that NP-1 formed complexes with VEGFR1 and VEGFR2, respectively, in cardiomyocytes. These findings suggest that NP-1 should play beneficial role in heart failure. PMID- 18381068 TI - Consequences of ions and pH on the supramolecular organization of sphingomyelin and sphingomyelin/cholesterol bilayers. AB - For drug delivery purpose the anticancer drug S12363 was loaded into ESM/Chol liposomes using either a pH or an ammonium gradient. Association between the drug and the liposome depends markedly on the liposome membrane structure. Thus, ESM and ESM/Chol bilayer organization had been characterized by coupled DSC and XRDT as a function of both cholesterol concentration and aqueous medium composition. ESM bilayers exhibited a ripple lamellar gel phase P(beta') below the melting temperature and adopted a L(beta)-like gel phase upon Chol insertion. Supramolecular organization of ESM and ESM/Chol bilayers was not modified by citrate buffer or ammonium sulfate solution whatever the pH (3< or = pH < or =7). Nevertheless, in ESM bilayer, ammonium sulfate salt induced a peculiar organization of head groups, leading to irregular d-spacing and weakly correlated bilayers. Moreover, in the presence of salts, a weakening of van der Waals attraction forces was seen and led to a swelling of the water layer. PMID- 18381069 TI - Photophysics of a beta-carboline based non-ionic probe in anionic and zwitterionic liposome membranes. AB - Interaction of a biologically active beta-carboline based non-ionic probe, 3 acetyl-4-oxo-6,7-dihydro-12H indolo-[2,3-a] quinolizine (AODIQ), with the liposomal vesicles of dimyristoyl-l-alpha-phosphatidylcholine (DMPC) and dimyristoyl-l-alpha-phosphatidylglycerol (DMPG) has been demonstrated using steady-state and time-resolved fluorescence and fluorescence anisotropy techniques. Polarity sensitive intramolecular charge transfer of AODIQ shows a large hypsochromic shift along with an enhancement in the fluorescence quantum yield and fluorescence lifetime in the bilayer membranes compared to those in aqueous buffer solution. Polarity of the immediate vicinity of the probe in the lipid environments has been determined. The fluorometric, quenching and micropolarity determination studies reveal that the fluorophore penetrates deeper in the zwitterionic DMPC membrane compared to the anionic DMPG vesicle. Enhancement in the rotational relaxation time of AODIQ in liposomal membranes suggests that the fluorophore exists in motionally restricted environments. PMID- 18381070 TI - Thermal acoustic radiation from multilamellar vesicles in lipid phase transition. AB - A new acoustical method for the investigation of lipid phase transition is introduced based on the measurement of the thermal acoustic radiation (TAR) inherent in lipids. The TAR of multilamellar vesicles from dipalmitoylphosphatidylcholine (DPPC) and dimyristoylphosphatidylcholine (DMPC) was measured in the megahertz range and the variations in the radiation intensity during the lipid phase transition were recorded. Two types of variations are possible: if the temperature of the vesicles decreases (in the process of transition from the liquid crystalline state to the gel state) then the TAR intensity increases, and if the temperature increases (in the reverse transition) then the TAR intensity decreases. These effects are connected with an increase in the ultrasonic absorption in the vesicles under lipid phase transition. Basing on the results of the TAR investigation, a new theoretical estimate has been developed of the variation in the absorption coefficient during the lipid phase transition. In this estimate, the variation is equated to the ratio of the phase transition entropy to the gas constant. PMID- 18381071 TI - Protein arginine (N)-methyl transferase 7 (PRMT7) as a potential target for the sensitization of tumor cells to camptothecins. AB - PRMT7 belongs to the protein arginine methyl-transferases family. We show that downregulation of PRMT7alpha and beta isoforms in DC-3F hamster cells was associated with increased sensitivity to the Top1 inhibitor camptothecin (CPT). This effect was not due to a change in Top1 contents or catalytic activity, or to a difference in the reversal of DNA breaks. Overexpression of PRMT7alpha and beta in DC-3F cells had no effect on CPT sensitivity, whereas it conferred a resistance to DC-3F/9-OH-E cells for which both isoforms are reduced by two- to three-fold as compared to DC-3F parental cells. Finally, downregulation of the human PRMT7 could also sensitize HeLa cells to CPT, suggesting that it could be used as a target to potentiate CPT derivatives. PMID- 18381072 TI - Dynamics of Cdc42 network embodies a Turing-type mechanism of yeast cell polarity. AB - Complex biochemical networks can be understood by identifying their principal regulatory motifs and mode of action. We model the early phase of budding yeast cellular polarization and show that the biochemical processes in the presumptive bud site comprise a Turing-type mechanism. The roles of the prototypical activator and substrate are played by GTPase Cdc42 in its active and inactive states, respectively. We demonstrate that the nucleotide cycling of Cdc42 converts cellular energy into a stable cluster of activated Cdc42. This energy drives a continuous membrane-cytoplasmic exchange of the cluster components to counteract diffusive spread of the cluster. This exchange explains why only one bud forms per cell cycle, because the winner-takes-all competition of candidate sites inevitably selects a single site. PMID- 18381073 TI - Osteoactivin is a novel osteoclastic protein and plays a key role in osteoclast differentiation and activity. AB - This study presents gene expression, protein expression, and in situ immunohistochemical evidence that osteoclasts express high levels of osteoactivin (OA), which had previously been reported to be an osteoblast-specific protein in bone. OA expression in osteoclasts was up-regulated upon receptor activator of NFkappaB ligand-induced differentiation. Suppression of functional activity of OA with neutralizing antibody reduced cell size, number of nuclei, fusion, and bone resorption activity of osteoclasts. OA was co-immunoprecipitated with integrin beta3 and beta1, indicating that OA co-localizes with integrin beta3 and/or beta1 in a hetero-polymeric complex in osteoclasts. These findings indicate that OA is a novel osteoclastic protein and plays a role in osteoclast differentiation and/or activity. PMID- 18381074 TI - Small mitochondrial ARF (smARF) is located in both the nucleus and cytoplasm, induces cell death, and activates p53 in mouse fibroblasts. AB - The ARF transcript produces two proteins, the full-length ARF, p19(ARF), and a short mitochondrial version, smARF. To explore the functional difference between the two, we generated GFP-fused expression vectors for each protein and introduced them into NIH3T3 murine fibroblasts, which sustains a global deletion in the INK4a locus but contains a functional p53 gene. GFP-p19ARF was located within the nucleolus as previously reported, whereas GFP-smARF was detected mainly in the nucleoplasm. GFP-smARF induced cell death although to a slightly lesser extent than p19ARF. GFP-smARF stabilized p53 thereby inducing expression of the target genes, MDM2 and p21. We suggest that smARF has functions other than mitochondria-mediated autophagy, and induces p53 expression and cell death via a novel mechanism. PMID- 18381075 TI - Establishment of the culture model system that reflects the process of terminal differentiation of connective tissue-type mast cells. AB - To understand physiological roles of tissue mast cells, we established a culture system where bone marrow-derived immature mast cells differentiate into the connective tissue-type mast cell (CTMC)-like cells through modifying the previous co-culture system with Swiss 3T3 fibroblasts. Our system was found to reproducibly mimic the differentiation of CTMCs on the basis of several criteria, such as granule maturation and sensitivity to cationic secretagogues. The gene expression profile obtained by the microarray analyses was found to reflect many aspects of the differentiation. Our system is thus helpful to gain deeper insights into terminal differentiation of CTMCs. PMID- 18381076 TI - Stimulation- and palmitoylation-dependent changes in oligomeric conformation of serotonin 5-HT1A receptors. AB - In the present study we analyzed the oligomerization state of the serotonin 5 HT1A receptor and studied oligomerization dynamics in living cells. We also investigated the role of receptor palmitoylation in this process. Biochemical analysis performed in neuroblastoma N1E-115 cells demonstrated that both palmitoylated and non-palmitoylated 5-HT1A receptors form homo-oligomers and that the prevalent receptor species at the plasma membrane are dimers. A combination of an acceptor-photobleaching FRET approach with fluorescence lifetime measurements verified the interaction of CFP- and YFP-labeled wild-type as well as acylation-deficient 5-HT1A receptors at the plasma membrane of living cells. Using a novel FRET technique based on the spectral analysis we also confirmed the specific nature of receptor oligomerization. The analysis of oligomerization dynamics revealed that apparent FRET efficiency measured for wild-type oligomers significantly decreased in response to agonist stimulation, and our combined results suggest that this decrease was mediated by accumulation of FRET-negative complexes rather than by dissociation of oligomers to monomers. In contrast, the agonist-mediated decrease of FRET signal was completely abolished in oligomers composed by non-palmitoylated receptor mutants, demonstrating the importance of palmitoylation in modulation of the structure of oligomers. PMID- 18381077 TI - Direct protein-protein interaction of 11beta-hydroxysteroid dehydrogenase type 1 and hexose-6-phosphate dehydrogenase in the endoplasmic reticulum lumen. AB - Hexose-6-phosphate dehydrogenase (H6PDH) has been shown to stimulate 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1)-dependent local regeneration of active glucocorticoids. Here, we show that coexpression with H6PDH results in a dramatic shift from 11beta-HSD1 oxidase to reductase activity without affecting the activity of the endoplasmic reticular enzyme 17beta-HSD2. Immunoprecipitation experiments revealed coprecipitation of H6PDH with 11beta-HSD1 but not with the related enzymes 11beta-HSD2 and 17beta-HSD2, suggesting a specific interaction between H6PDH and 11beta-HSD1. The use of the 11beta-HSD1/11beta-HSD2 chimera indicates that the N-terminal 39 residues of 11beta-HSD1 are sufficient for interaction with H6PDH. An important role of the N-terminus was indicated further by the significantly stronger interaction of 11beta-HSD1 mutant Y18-21A with H6PDH compared to wild-type 11beta-HSD1. The protein-protein interaction and the involvement of the N-terminus of 11beta-HSD1 were confirmed by Far-Western blotting. Finally, fluorescence resonance energy transfer (FRET) measurements of HEK-293 cells expressing fluorescently labeled proteins provided evidence for an interaction between 11beta-HSD1 and H6PDH in intact cells. Thus, using three different methods, we provide strong evidence that the functional coupling between 11beta-HSD1 and H6PDH involves a direct physical interaction of the two proteins. PMID- 18381078 TI - Functional role of N-glycosylation from ADAM10 in processing, localization and activity of the enzyme. AB - A disintegrin and metalloprotease 10 (ADAM10) is a type I transmembrane glycoprotein with four potential N-glycosylation sites (N267, N278, N439 and N551), that cleaves several plasma membrane proteins. In this work, ADAM10 was found to contain high-mannose and complex-type glycans. Individual N glycosylation site mutants S269A, T280A, S441A, T553A were constructed, and results indicated that all sites were occupied. T280A was found to accumulate in the endoplasmic reticulum as the non-processed precursor of the enzyme. Furthermore, it exhibited only residual levels of metalloprotease activity in vivo towards the L1 cell adhesion molecule, as well as in vitro, using a ProTNF alpha peptide as substrate. S441A showed increased ADAM10 susceptibility to proteolysis. Mutation of N267, N439 and N551 did not completely abolish enzyme activity, however, reduced levels were found. ADAM10 is sorted into secretory vesicles, the exosomes. Here, a fraction of ADAM10 from exosomes was found to contain more processed N-linked glycans than the cellular enzyme. In conclusion, N-glycosylation is crucial for ADAM10 processing and resistance to proteolysis, and results suggest that it is required for full-enzyme activity. PMID- 18381079 TI - A trifunctional enzyme with glutathione S-transferase, glutathione peroxidase and superoxide dismutase activity. AB - Superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione S transferase (GST) and glutathione reductase (GR) play crucial roles in balancing the production and decomposition of reactive oxygen species (ROS) in living organisms. These enzymes act cooperatively and synergistically to scavenge ROS, as not one of them can singlehandedly clear all forms of ROS. In order to imitate the synergy of the enzymes, we designed and generated a recombinant protein, which comprises of a Schistosoma japonicum GST (SjGST) and a bifunctional 35-mer peptide with SOD and GPX activities. The engineered protein demonstrated SOD, GPX and GST activities simultaneously. This trifunctional enzyme with SOD, GPX and GST activities is expected to be the best ROS scavenger. PMID- 18381080 TI - Thiyl radicals react with nitric oxide to form S-nitrosothiols with rate constants near the diffusion-controlled limit. AB - A possible route to S-nitrosothiols in biology is the reaction between thiyl radicals and nitric oxide. D. Hofstetter et al. (Biochem. Biophys. Res. Commun.360:146-148; 2007) claimed an upper limit of (2.8+/-0.6)x10(7) M(-1)s(-1) for the rate constant between thiyl radicals derived from glutathione and nitric oxide, and it was suggested that under physiological conditions S-nitrosation via this route is negligible. In the present study, thiyl radicals were generated by pulse radiolysis, and the rate constants of their reactions with nitric oxide were determined by kinetic competition with the oxidizable dyes 2,2'-azino-bis(3 ethylbenzothiazoline-6-sulfonate) and a phenothiazine. The rate constants for the reaction of nitric oxide with thiyl radicals derived from glutathione, cysteine, and penicillamine were all in the range (2-3) x10(9) M(-1)s(-1), two orders of magnitude higher than the previously reported estimate in the case of glutathione. Absorbance changes on reaction of thiyl radicals with nitric oxide were consistent with such high reactivity and showed the formation of S nitrosothiols, which was also confirmed in the case of glutathione by HPLC/MS. These rate constants imply that formation of S-nitrosothiols in biological systems from the combination of thiyl radicals with nitric oxide is much more likely than claimed by Hofstetter et al. PMID- 18381081 TI - Rescue of mutant alpha-galactosidase A in the endoplasmic reticulum by 1 deoxygalactonojirimycin leads to trafficking to lysosomes. AB - Active-site-specific chaperone therapy for Fabry disease is a genotype-specific therapy using a competitive inhibitor, 1-deoxygalactonojirimycin (DGJ). To elucidate the mechanism of enhancing alpha-galactosidase A (alpha-Gal A) activity by DGJ-treatment, we studied the degradation of a mutant protein and the effect of DGJ in the endoplasmic reticulum (ER). We first established an in vitro translation and translocation system using rabbit reticulocyte lysates and canine pancreas microsomal vesicles for a study on the stability of mutant alpha-Gal A with an amino acid substitution (R301Q) in the ER. R301Q was rapidly degraded, but no degradation of wild-type alpha-Gal A was observed when microsomal vesicles containing wild-type or R301Q alpha-Gal A were isolated and incubated. A pulse chase experiment on R301Q-expressing TgM/KO mouse fibroblasts showed rapid degradation of R301Q, and its degradation was blocked by the addition of lactacystin, indicating that R301Q was degraded by ER-associated degradation (ERAD). Rapid degradation of R301Q was also observed in TgM/KO mouse fibroblasts treated with brefeldin A, and the amount of R301Q enzyme markedly increased by pretreatment with DGJ starting 12 h prior to addition of brefeldin A. The enhancement of alpha-Gal A activity and its protein level by DGJ-treatment was selectively observed in brefeldin A-treated COS-7 cells expressing R301Q but not in cells expressing the wild-type alpha-Gal A. Observation by immunoelectron microscopy showed that the localization of R301Q in COS-7 cells was in the lysosomes, not the ER. These data suggest that the rescue of R301Q from ERAD is a key step for normalization of intracellular trafficking of R301Q. PMID- 18381082 TI - Regulation of the roles of sphingosine 1-phosphate and its type 1 G protein coupled receptor in T cell immunity and autoimmunity. AB - The lipid mediator sphingosine 1-phosphate (S1P) and its type 1 G protein-coupled receptor (S1P1) affect mammalian immunity through alterations in thymocyte emigration, differentiation of T cell subsets, lymphocyte trafficking in lymphoid organs and other tissues, T cell-dendritic cell and T cell-B cell interactions, and cytokine generation. Recent attention to effects of the S1P-S1P1 axis on non migration functions of lymphocytes includes delineation of a role in terminal differentiation and survival of Th17 effector cells and adaptive Treg cells of the CD4 T cell constellation, and a greater understanding of interactions of the S1P-S1P1 axis with immune cytokines in lymphocyte survival and activities. This breadth of involvement of the S1P-S1P1 axis in immune responses that often are altered in immunological diseases has provided many opportunities for novel therapeutic interventions. A spectrum of pharmacological and immunochemical agents is available that alter immunity by affecting either tissue and fluid concentrations of S1P or levels of expression and signaling activities of S1P1. Such agents have so far been beneficial in the settings of autoimmunity and rejection of transplanted organs, and are likely to become valuable constituents of combined drug programs. PMID- 18381083 TI - Determinants for substrate phosphorylation by Dictyostelium myosin II heavy chain kinases A and B and eukaryotic elongation factor-2 kinase. AB - The alpha kinases are a widespread family of atypical protein kinases characterized by a novel type of catalytic domain. In this paper the peptide substrate recognition motifs for three alpha kinases, Dictyostelium discoideum myosin heavy chain kinase (MHCK) A and MHCK B and mammalian eukaryotic elongation factor-2 kinase (eEF-2K), were characterized by incorporating amino acid substitutions into a previously identified MHCK A peptide substrate (YAYDTRYRR) (Luo X. et al. (2001) J. Biol. Chem. 276, 17836-43). A lysine or arginine in the P+1 position on the C-terminal side of the phosphoacceptor threonine (P site) was found to be critical for peptide substrate recognition by MHCK A, MHCK B and eEF 2K. Phosphorylation by MHCK B was further enhanced 8-fold by a basic residue in the P+2 position whereas phosphorylation by MHCK A was enhanced 2- to 4-fold by basic residues in the P+2, P+3 and P+4 positions. eEF-2K required basic residues in both the P+1 and P+3 positions to recognize peptide substrates. eEF-2K, like MHCK A and MHCK B, exhibited a strong preference for threonine as the phosphoacceptor amino acid. In contrast, the Dictyostelium VwkA and mammalian TRPM7 alpha kinases phosphorylated both threonine and serine residues. The results, together with a phylogenetic analysis of the alpha kinase catalytic domain, support the view that the metazoan eEF-2Ks and the Dictyostelium MHCKs form a distinct subgroup of alpha kinases with conserved properties. PMID- 18381084 TI - Kinetic study of the thermal inactivation of cholinesterase enzymes immobilized in solid matrices. AB - The thermal inactivation of immobilized cholinesterase enzymes (ChE) in solid matrices where the protein unfolding is blocked was studied, thus enabling investigation of the kinetics of the inactivation process directly from the native structure to the inactivated state. The thermal inactivation of butyrylcholinesterase (BChE), recombinant human acetylcholinesterase (rHuAChE), and eel acetylcholinesterase (AChE) enzymes was studied in dry films composed of poly(vinyl pyrollidone) (PVP), bovine serum albumin (BSA) and trehalose at 60 degrees -120 degrees C. The kinetics follows a bi-exponential decay equation representing a combination of fast and slow processes. The activation enthalpy DeltaH(#) and the activation entropy DeltaS(#) for each of the three enzymes have been evaluated. The values of DeltaH(#) for the fast process and for the slow process of BChE are 33+/-3, and 28+/-2 kcal/mol, respectively, and the values of DeltaS(#) are 0.84+/-0.04, and -18.2+/-0.5 cal/deg, respectively. The appropriate value of DeltaH(#) for rHuAChE is 26+/-2 Kcal/mol, for both processes and the values of DeltaS(#) are -17.6+/-0.9, and -23.0+/-0.9 cal/deg, respectively. Similarly, the values of DeltaH(#) for eelAChE are 30+/-3, 31+/-1 kcal/mol, and the values of DeltaS(#) are -6.7+/-0.5, -9.1+/-0.2 cal/deg respectively. PMID- 18381086 TI - Pediatric hematology and pediatric oncology. PMID- 18381087 TI - Decision analysis in pediatric hematology. AB - Decision analysis is a simulation, model-based research technique in which investigators combine information from a variety of sources to create a mathematical model representing a clinical decision. This tool can be used to address many clinical dilemmas in pediatric hematology for which traditional clinical trials are unfeasible or impossible. This article outlines the basic steps of performing and analyzing a decision analysis tree and describes several decision analyses published in the field of pediatric hematology and how to evaluate and judge the decision analysis literature. PMID- 18381085 TI - MicroRNA-206: the skeletal muscle-specific myomiR. AB - MicroRNAs (miRNAs) are a class of non-coding RNAs involved in post transcriptional gene silencing. A small number of striated muscle-specific miRNAs have been identified and shown to have an important role in myogenesis, embryonic muscle growth and cardiac function and hypertrophy. One of these myomiRs (myo=muscle+miR=miRNA), miR-206, is unique in that it is only expressed in skeletal muscle. The purpose of this review is to discuss what is currently known about miR-206 and its function in myogenesis as well as propose potential new roles for miR-206 in skeletal muscle biology. The review is also intended to serve as a comprehensive resource for miR-206 with the hope of encouraging further research on the role of miR-206 in skeletal muscle. PMID- 18381088 TI - Venous thromboembolism in children. AB - With improved pediatric survival from serious underlying illnesses, greater use of invasive vascular procedures and devices, and a growing awareness that vascular events occur among the young, venous thromboembolism (VTE) increasingly is recognized as a critical pediatric concern. This review provides background on etiology and epidemiology in this disorder, followed by an in-depth discussion of approaches to the clinical characterization, diagnostic evaluation, and management of pediatric VTE. Prognostic indicators and long-term outcomes are considered, with emphasis on available evidence underlying current knowledge and key questions for further investigation. PMID- 18381089 TI - Pediatric arterial ischemic stroke. AB - Arterial ischemic stroke (AIS) is a rare disorder in children. Research suggests that risk factors, outcomes, and presentation are different from those of adult stroke. In particular, prothrombotic abnormalities and large vessel arteriopathies that are nonatherosclerotic seem to play a large role in the pathogenesis of childhood AIS. This review examines the epidemiology and etiologies of neonatal and childhood AIS and provides a detailed discussion of approaches to the clinical characterization, diagnostic evaluation, and management. Long-term outcomes of recurrent AIS and neuromotor, speech, cognitive, and behavioral deficits are considered. Emphasis is on evidence underlying current knowledge and key questions for further investigation. PMID- 18381090 TI - The role of the hematologist/oncologist in the care of patients with vascular anomalies. AB - Pediatric hematologist/oncologists have a critical role in the diagnosis and management of patients who have complex vascular anomalies. They provide the clinical and medical skills needed to diagnose, treat, and manage these patients. Hematologist/oncologists also provide support for clinical trials and drug development to further treatment options for these patients. PMID- 18381091 TI - Advances in hemophilia: experimental aspects and therapy. AB - This article describes recent clinical and research advances in hemophilia therapy. Different prophylactic regimens for the management of severe hemophilia are described along with the use of adjuvant treatment options to achieve hemostasis. The safety and efficacy of radionuclide synovectomy with phosphorus 32-sulfur colloid to treat existing joint arthropathy also are described. The development of inhibitors to factor VIII or IX remains a challenge for hemophilia care and recent approaches to achieve immune tolerance induction are discussed. Finally, recent advances in hemophilia are mentioned, including the role of iron, inflammation, and angiogenesis in the pathogenesis of hemophilic arthopathy. PMID- 18381092 TI - Von Willebrand disease. AB - von Willebrand disease is a common inherited bleeding disorder and many cases are diagnosed in childhood. It has a negative impact on the quality of life of affected individuals; therefore, it is important that the condition be recognized and diagnosed. This article reviews the pathophysiology of the condition, the current classification scheme, and the available treatments, highlighting issues specific to the pediatric population. PMID- 18381093 TI - Childhood immune thrombocytopenic purpura: diagnosis and management. AB - Immune thrombocytopenic purpura (ITP) is an autoimmune disorder characterized by a low circulating platelet count caused by destruction of antibody-sensitized platelets in the reticuloendothelial system. ITP can be classified as childhood versus adult, acute versus chronic, and primary versus secondary. Persistence of thrombocytopenia defines the chronic form of the disorder. Secondary causes of ITP include collagen vascular disorders, immune deficiencies, and some chronic infections. This review focuses on the diagnosis and management of children who have acute and chronic ITP. Emphasis is placed on areas of controversy and new therapies. PMID- 18381094 TI - Blood component therapy. AB - Blood component transfusion is integral in the treatment of infants and children by pediatricians, surgeons, intensivists, and hematologists/oncologists. Technologic advances in blood collection, separation, anticoagulation, and preservation have resulted in component preparation of red blood cells, platelets, white blood cells, and plasma, which are superior to whole blood used in the past. Advances in donor selection, infectious disease testing, leukoreduction filters, and gamma irradiation have made products safer. Physicians prescribing blood components should have a basic understanding of indications (and contraindications) and be cognizant of methods of preparation, proper storage conditions, and requirements for modification of blood products to prevent potential adverse effects. PMID- 18381095 TI - Update on thalassemia: clinical care and complications. AB - beta-Thalassemia, originally named Cooley anemia, is an inherited blood disease. Various types of thalassemia are inherited anemias caused by mutations at the globin gene loci on chromosomes 16 and 11, affecting the production of alpha- or beta-globin protein, respectively. The combination of early diagnosis, improvements in monitoring for organ complications, and advances in supportive care have enabled many patients who have severe thalassemia syndromes to live productive, active lives well into adulthood. PMID- 18381096 TI - Oral iron chelators. AB - Effective chelation therapy can prevent or reverse organ toxicity related to iron overload, yet cardiac complications and premature death continue to occur, largely related to difficulties with compliance in patients who receive parenteral therapy. The use of oral chelators may be able to overcome these difficulties and improve patient outcomes. A chelator's efficacy at cardiac and liver iron removal and side-effect profile should be considered when tailoring individual chelation regimens. Broader options for chelation therapy, including possible combination therapy, should improve clinical efficacy and enhance patient care. PMID- 18381097 TI - Hydroxyurea for children with sickle cell disease. AB - Hydroxyurea therapy offers promise for ameliorating the clinical course of children with sickle cell disease (SCD). Hydroxyurea is a prototypic therapeutic option; it can be administered with minimal side effects, has a relatively wide therapeutic window, and has mechanisms of action that address pathophysiologic pathways of sickling, vaso-occlusion, hemolysis, and organ damage. There are limited data regarding hydroxyurea's ability to prevent or diminish organ dysfunction, and the long-term risks of hydroxyurea therapy remain incompletely defined. Although clinical trials are underway to address long-term issues, hydroxyurea remains an effective but underutilized therapy for SCD. PMID- 18381098 TI - Partial splenectomy for hereditary spherocytosis. AB - The significant risks associated with total splenectomy have led to interest in the use of partial splenectomy as an alternative surgical therapy for children who have congenital hemolytic anemia. Partial splenectomy is designed to remove enough spleen to gain desired hematologic outcomes while preserving splenic immune function. Although preliminary data demonstrate successful laboratory and clinical outcomes after partial splenectomy in various congenital hemolytic anemias, conclusive data comparing the efficacy of partial splenectomy to total splenectomy are not reported. Based on preliminary data, a definitive clinical trial of partial splenectomy in children who have severe congenital hemolytic anemia may be warranted. PMID- 18381100 TI - Monoclonal antibodies in lymphoma: the first decade. AB - The past decade has seen improvements in overall survival (OS) for patients with the two most common lymphoma histologies: diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL). In FL, at least four independent datasets have confirmed these survival improvements. The monoclonal antibody rituximab has significantly contributed to these improved therapeutic outcomes. This issue will explore the impact of monoclonal antibody therapy on outcome in indolent lymphoma, and detail how this improved outcome has changed clinical practice. Furthermore, the role of monoclonal antibodies in maintenance regimens and the main indications for radiolabeled antibodies will be discussed in detail. Finally, possible future developments in the field will be proposed, including the use of monoclonal antibodies in ablative transplantation and in the treatment of leukemias. PMID- 18381101 TI - Monoclonal antibodies for the treatment of hematologic malignancies: schedule and maintenance therapy. AB - In the last decade rituximab, alemtuzumab, and gemtuzumab ozogamicin (GO) have been used to treat patients with hematologic malignancies. Their efficacy and safety are now well established. Since their preclinical development, many studies have been performed to optimize dose and schedule. Rituximab is usually given at 375 mg/m(2), a dose that shows activity and little toxicity. It is normally administered as single agent or in combination with chemotherapy to induce remission in B-cell neoplasias. Moreover, given its low toxicity and long half-life, rituximab also can be used as maintenance therapy. Alemtuzumab is administered with a schedule of 30 mg, three times per week, after an initial dose escalation in the first week, showing activity against chronic lymphocytic leukemia (B-CLL) and some T-cell neoplasias. GO is administered at a dose of 9 mg/m(2) at 2-week intervals for two doses; it is the first monoclonal antibody approved for the treatment of relapsed or refractory CD33(+) acute myeloid leukemia (AML). PMID- 18381102 TI - Antibody and immunomodulatory agents in the treatment of indolent non-Hodgkin's lymphoma. AB - Immunomodulatory agents, including cytokines, CpG oligonucleotides, and anti idiotype vaccines have properties that suggest they have the ability to augment rituximab in the treatment of non-Hodgkin's lymphoma (NHL). Although several clinical trials have shown promising results, no randomized trials of reasonable size have been reported to date, limiting the ability to discern whether combinations of immunomodulatory agents with rituximab impact clinical outcome. Until such trials are mature, we do not recommend using these agents in combination outside of the research setting. PMID- 18381103 TI - Chemotherapy combinations with monoclonal antibodies in non-Hodgkin's lymphoma. AB - Although the use of monoclonal antibodies as single agents has had a tremendous impact on the care of patients with non-Hodgkin's lymphoma (NHL), the greatest benefit has been generated by the addition of monoclonal antibodies to conventional cytotoxic chemotherapy. Rituximab is the monoclonal antibody responsible for all clinical improvement noted to date. The addition of rituximab to cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy (R-CHOP regimen) improves the response rate, progression-free survival (PFS), and overall survival (OS) in diffuse large B-cell lymphoma (DLBCL). Adding rituximab to CHOP chemotherapy improves response rates and PFS in mantle cell lymphoma (MCL). Finally, the addition of rituximab to a variety of chemotherapy regimens improves the response rates, PFS, and OS in follicular lymphoma (FL). Several other (epratuzumab, bevacizumab, alemtuzumab) monoclonal antibody-chemotherapy combinations are currently under study in NHL. This review will summarize the data supporting the addition of rituximab to chemotherapy in NHL and discuss preliminary data regarding the use of other monoclonal antibodies in combination with chemotherapy. PMID- 18381104 TI - Antibody therapy for chronic lymphocytic leukemia. AB - The introduction of the monoclonal antibodies rituximab (anti-CD20) and alemtuzumab (anti-CD52) has revolutionized the treatment of chronic lymphocytic leukemia (CLL). Both antibodies were first studied as single agents in relapsed CLL, but rituximab is increasingly used in combination chemoimmunotherapy regimens in previously untreated patients. Phase II studies demonstrated that the addition of rituximab to fludarabine-based chemotherapy improves complete response (CR) rates and prolongs progression-free survival (PFS), but a long-term survival benefit has not been shown. Alemtuzumab is less commonly used, due to the greater likelihood of infusion toxicity, as well as hematologic and immune toxicities. Subcutaneous (SC) administration significantly reduces infusion toxicity, but hematologic and infectious complications, most notably cytomegalovirus (CMV) reactivation, still occur with SC dosing. Alemtuzumab's unique clinical properties include its clinical activity in relapsed CLL patients with del(17p13) and its ability to eradicate minimal residual disease (MRD) in bone marrow. Its use as consolidation therapy to eradicate MRD after nucleoside analog therapy is under active study. Several investigational monoclonal antibodies are in preclinical or clinical studies, most notably lumiliximab (anti CD23) and ofatumumab (HuMax CD20), and are briefly discussed in this review. PMID- 18381105 TI - Antibody therapy for acute myeloid leukemia. AB - Due to the high rate of relapse in younger patients and the overall poor outcome in older patients, novel therapies are needed for the treatment of acute myeloid leukemia (AML). Monoclonal antibodies have become an important treatment modality in cancer therapy. Gemtuzumab ozogamicin (GO), an anti-CD33 immunoconjugate, was approved by the US Food and Drug Administration (FDA) for the treatment of elderly patients with relapsed AML who are not candidates for standard chemotherapy. Single-agent GO and combinations with standard chemotherapeutics have been explored extensively in this disease. Hepatotoxicity and delayed myelosuppression have been dose-limiting. Its toxicity profile is reduced with decreased doses of GO and even by administering only a single infusion. In patients with acute promyelocytic leukemia (APL), the addition of GO can produce molecular remissions and is well tolerated. Targeted immunotherapy with GO for treatment of AML has produced remissions. In order to reduce toxicity and improve efficacy, its optimal dose and schedule and pairing with other standard chemotherapeutic agents need to be defined better in large clinical trials. PMID- 18381106 TI - Non-myeloablative radioimmunotherapy for non-Hodgkin's lymphoma. AB - Iodine 131-tositumomab and ibritumomab tiuxetan, radioimmunoconjugates that target the CD20 antigen, have been approved in the United States for use in relapsed or refractory, indolent, and transformed B-cell lymphomas. These well tolerated agents have among the highest single-agent activity observed for these histologies. Furthermore, emerging data suggest a role for radioimmunotherapy (RIT) as upfront therapy of indolent lymphoma, with expanding utility in the treatment of more aggressive histologies. This review will summarize the key trials that led to approval of both (131)I-tositumomab and ibritumomab tiuxetan, discuss recent publications on re-treatment and therapy of de novo indolent lymphoma and aggressive histologies, and conclude with new topics in RIT. PMID- 18381109 TI - Obstetrics and gynecology for the general surgeon. Foreword. PMID- 18381108 TI - Novel and engineered anti-B-cell monoclonal antibodies for non-Hodgkin's lymphoma. AB - Over the past decade, the safety and efficacy of the anti-CD20 antibody rituximab has resulted in its use in virtually all patients with B-cell non-Hodgkin's lymphoma (NHL). Unfortunately, many patients who initially benefit from rituximab develop resistance while others may never respond. Both the successes and limitations of rituximab have heralded an explosion in research and development of novel monoclonal antibodies. Strategies employed to improve upon rituximab have included developing antibodies to target new epitopes of CD20 and new antigens, humanizing or creating fully human antibodies, and engineering antibodies with a potentially greater capacity for interaction with the host immune system. Each of these strategies has shown varying degrees of preclinical and clinical success. In this review we discuss the rationale for various strategies and report results from clinical trials employing these agents. PMID- 18381107 TI - Radioimmunotherapy-based conditioning regimens for stem cell transplantation. AB - Radioimmunotherapy (RIT) combines the mechanism of action and targeting capability of monoclonal antibodies with the tumoricidal effect of radiation and has shown promising results in the treatment of various hematologic malignancies. Based on RIT's efficacy and safety profile, many investigators have evaluated its use in transplant conditioning regimens with the goal of improving long-term disease control with limited toxicity. In lymphoma, two basic transplant approaches targeting CD20 have emerged: (1) myeloablative doses of RIT with or without chemotherapy, and (2) standard nonmyeloablative doses of RIT combined with high-dose chemotherapy. Myeloablative RIT has been shown to be feasible and efficacious using escalated doses of iodine 131-tositumomab, yttrium 90 ibritumomab tiuxetan, and (131)I-rituximab with or without chemotherapy followed by autologous stem cell transplant (ASCT). The second approach predominantly has used standard doses of (90)Y-ibritumomab tiuxetan or (131)I-tositumomab plus BEAM chemotherapy (carmustine [BCNU], etoposide, cytarabine, melphalan) followed by ASCT. RIT targeting CD45, CD33, and CD66 prior to allogeneic transplantation also has been evaluated for the treatment of acute leukemia. Overall RIT-based transplant conditioning for lymphoma and leukemia has been shown to be safe, effective, and feasible with ongoing randomized trials currently underway to definitively establish its place in the treatment of hematologic malignancies. PMID- 18381110 TI - Obstetrics and gynecology for the general surgeon. Preface. PMID- 18381111 TI - Surgical exposure and anatomy of the female pelvis. AB - Female pelvic anatomy encompasses the reproductive, urologic, and gastrointestinal systems. Knowledge of the inherent relations between these organ systems, as well as the ability to develop pelvic spaces, will enable the surgeon to approach pelvic pathology confidently. This article highlights basic anatomy of the female pelvis and emphasizes points of caution during pelvic surgery, as well as reviews the essential principles of pelvic support. PMID- 18381112 TI - Benign gynecologic conditions. AB - Benign gynecologic conditions constitute the majority of the general gynecologist's practice. Along with health maintenance examinations, contraceptive management, family planning issues, and concerns about incontinence, the gynecologic conditions for which patients commonly present include adnexal masses, leiomyomata, endometriosis, and pelvic inflammatory disease. This article addresses each of these last four entities and incorporates a discussion of their etiologies, clinical presentations, keys to diagnosis, and the various treatment options available. PMID- 18381113 TI - Gynecologic emergencies. AB - Gynecologic emergencies are relatively common and include ectopic pregnancies, adnexal torsion, tubo-ovarian abscess, hemorrhagic ovarian cysts, gynecologic hemorrhage, and vulvovaginal trauma. The purpose of this article is to provide a concise review of these emergencies, focusing on the evaluation and treatment options for the patient. In many cases, other causes of an acute abdomen are in the differential diagnosis. Understanding the tenets of diagnosis helps the surgeon narrow the etiology and guide appropriate treatment. PMID- 18381114 TI - Ovarian cancer. AB - Ovarian cancer ranks fifth in cancer deaths among women and causes more deaths than any other gynecologic malignancy. It is estimated that 22,430 new cases will be diagnosed and 15,280 deaths will be attributed to ovarian cancer in the United States in 2007. Despite initial responses to surgery and chemotherapy in up to 80% of cases, more than 75% of affected women ultimately die from complications of disease progression. PMID- 18381115 TI - Gynecologic malignancies. AB - The pioneers in the field of gynecologic oncology set out to establish an evidence-based approach to the care of women with gynecologic cancer, combining the modalities of surgery, chemotherapy, and radiation. Quality of life has become the cornerstone of care for these patients, in addition to advancing survival through surgical technology, collaborative research trials, and molecular approaches to early diagnosis and management. This article addresses the epidemiology, screening, preventive strategies, diagnosis, staging, surgical care, adjuvant therapy, prognosis, and recurrence management of three common gynecologic malignancies encountered in the operating arena: endometrial, cervical, and vulvar cancer. PMID- 18381116 TI - Gynecologic laparoscopy. AB - Gynecologic laparoscopy is an evolving discipline. Major advances in technology have facilitated the transition from traditional "open" surgery to minimally invasive procedures. This article reviews the basic tenets of gynecologic laparoscopy: pelvic anatomy, general procedures, laparoscopic instruments, complications, and common gynecologic surgeries. Highlighted procedures include adnexal surgery, laparoscopic hysterectomy, and operations for female malignancies. PMID- 18381117 TI - Complications of gynecologic surgery. AB - Complications during gynecologic surgery result from the proximity of the uterus and ovaries to other critical pelvic structures. These structures include the urinary tract, bowel, nerves, and vasculature. Knowledge of pelvic anatomy is important when performing these procedures and is critical in cases of altered anatomy from adhesive disease and during intraoperative hemorrhage. Recognition and repair of an unintended injury gives the best chance for minimizing sequelae from these complications. PMID- 18381119 TI - Physiologic changes in pregnancy. AB - Pregnancy is associated with physiologic changes that affect virtually every organ system in the mother. These physiologic changes allow the parturient to support the growing uterus and fetus and to withstand labor and the postpartum course. Because the gravida may require surgery while pregnant, it is important to have an appreciation of these physiologic changes. These maternal adaptive responses have significant implications for anesthetic, operative, and perioperative management. PMID- 18381118 TI - CT, MRI, PET, PET/CT, and ultrasound in the evaluation of obstetric and gynecologic patients. AB - The role of imaging in obstetrics and gynecology has undergone a revolution over the past few decades. Well-established methods such as endovaginal ultrasound have had a central role in the evaluation of nongravid patients with pelvic pain, as well as in the workup for ectopic pregnancy and evaluation of adnexal masses. Additional tools include MRI in the evaluation of appendicitis and other potentially surgical conditions in pregnant patients and MRI and CT in the evaluation of surgical complications. Newer tools in the radiology armamentarium include PET scanning which, alongside MRI and CT, are often helpful in staging gynecologic malignancy. The role of imaging in the obstetric and gynecology patient will continue to change as new modalities and techniques are introduced. PMID- 18381120 TI - Surgical diseases presenting in pregnancy. AB - Surgical diseases frequently arise in gravid patients, with an estimated 1 in 500 pregnancies requiring nonobstetric surgical intervention. Evaluation and treatment generally are the same as for the nonpregnant patient, but pregnancy may mask some of the typical presenting symptoms, leading to delayed diagnosis. This article highlights some of the more common surgical diseases that may present during pregnancy, including appendicitis, biliary diseases, bowel obstruction, hemorrhoids, inflammatory bowel disease, and malignancies. PMID- 18381121 TI - Trauma and surgical emergencies in the obstetric patient. AB - Pregnancy always must be considered when evaluating a female trauma victim of reproductive age. When managing the pregnant trauma victim, one must optimize the well-being of two patients, but the health of the mother is of paramount importance. Rapid assessment, treatment, and transport are critical to optimizing maternal and fetal outcome. Evaluation must be performed with an understanding of the physiologic changes that occur in pregnancy. These changes alter maternal response to trauma and require adaptations to care. PMID- 18381122 TI - Appropriate selection of biochemical targets is one of the most important driver of cost-effectiveness in the discovery of new medicines. Preface. PMID- 18381123 TI - The discovery of raltegravir, an integrase inhibitor for the treatment of HIV infection. PMID- 18381124 TI - Inhibitors of 11beta-hydroxysteroid dehydrogenase type 1. PMID- 18381125 TI - Alpha7 nicotinic acetylcholine receptor agonists and positive allosteric modulators. PMID- 18381126 TI - Small molecule bradykinin B1 receptor antagonists as potential therapeutic agents for pain. PMID- 18381127 TI - Histone deacetylase inhibitors: a novel class of anti-cancer agents on its way to the market. PMID- 18381128 TI - 5-HT2C ligands: recent progress. PMID- 18381130 TI - Isolated iliac artery aneurysms: a contemporary comparison of endovascular and open repair. AB - OBJECTIVE: Iliac artery aneurysms are rare but associated with significant morbidity and mortality when ruptured. This study compares recent open and endovascular repairs of iliac aneurysms at a single institution. METHODS: Patients were identified and charts reviewed using ICD-9 and CPT codes for iliac artery aneurysm and open or endovascular repair performed between January 2000 and January 2006. Baseline characteristics, procedure-related variables, and follow-up data were retrospectively reviewed. RESULTS: A total of 71 patients were treated with isolated iliac artery aneurysms. There were 19 open and 52 endovascular repairs. Seven presented with acute ruptures and were treated by open (4) or endovascular (3) repair. Preoperative comorbidities were similar between the two groups. Major perioperative (30 day) complications included three deaths in the open group from cardiovascular complications, all after ruptured aneurysm repair, and one death in the endovascular group (after rupture; one additional perioperative death occurred after 30 days due to colonic infarction) (P = NS). Postoperative complications were less frequent in the endovascular group, although this did not reach statistical significance. The mortality was 50% in the open group and 33% in the endovascular group for patients presenting with a ruptured aneurysm (P = NS). Transfusion requirement was significantly higher in the open group (47%) than in the endovascular group (6%) (P = .03). The mean follow-up was 20 +/- 5 months in the open group and 17 +/- 2 months in the endovascular group (P = NS). Long-term complications included two limb thromboses following repair with a bifurcated stent graft that were treated with thrombolysis plus stenting or a fem-fem bypass. Three endoleaks were identified on postop CT scans, all of which were successfully managed with endovascular techniques. There were no postoperative ruptures or aneurysm-related death. The mean postoperative length of stay was 5.2 +/- 2.3 days (open) and 1.3 +/- 1.0 days (endovascular) (P = .04). CONCLUSIONS: This is the first large, case control study comparing open vs endovascular repair of isolated iliac artery aneurysms. Endovascular repair of iliac artery aneurysms is safe and results in decreased length of stay, lower requirement for perioperative blood transfusion, and similar intermediate term outcomes as open repair. PMID- 18381131 TI - Short and midterm results with minimally invasive endovascular repair of acute and chronic thoracic aortic pathology. AB - OBJECTIVES: Endovascular management of both acute and chronic thoracic aortic pathology has emerged as an alternative to open surgery. We reviewed our single center experience with endovascular devices for the treatment of thoracic aortic pathology. METHODS: Between April 2000 and October 2007, 116 thoracic aortic stent grafts were placed to treat a variety of acute or chronic thoracic aortic lesions. Thirty-five percent of the cases were performed emergently. Sixty-five percent of the patients were male; the average age was 63.9 years (range 20-93 years). Indications for treatment were chronic degenerative aneurysms (n = 70), traumatic aortic disruption (n = 20), complicated dissection, intramural hematoma, or penetrating aortic ulcer (n = 14), pseudoaneurysm (n = 10), and Diverticulum of Kommerell (n = 2). Arch vessel revascularization (n = 32) or mesenteric debranching (n = 7) was performed in select cases. Devices used were industry-approved thoracic aortic devices (n = 80), aortic cuff extenders (n = 19), or custom made by the surgeon (n = 17). RESULTS: The 30-day death, stroke and paraplegia/paresis rates were 5.2%, 8.6%, and 2.6%, respectively. Arterial access complications requiring immediate operative repair occurred at a rate of 11.2% (n = 13). The endoleak rate requiring repeat intervention was 6.9% (n = 8). The delayed graft infection rate was 5.2% (n = 6), with four of these cases resulting in death. The mean follow-up is 15 months (range 1-78 months). Computed tomography angiograms were performed at 1, 6, and 12 months following the index procedure, and yearly thereafter. CONCLUSIONS: Endovascular therapy for acute and chronic thoracic aortic pathology is a viable alternative to open surgery with comparable operative morbidity and mortality. Midterm results suggest that endografts are durable, but require more secondary interventions and imaging surveillance than open reconstruction. PMID- 18381133 TI - Hybrid procedures for thoracoabdominal aortic aneurysms and chronic aortic dissections - a single center experience in 28 patients. AB - OBJECTIVE: We report our 6-year experience with the visceral hybrid procedure for high-risk patients with thoracoabdominal aortic aneurysms (TAAA) and chronic expanding aortic dissections (CEAD). METHODS: Hybrid procedure includes debranching of the visceral and renal arteries followed by endovascular exclusion of the aneurysm. A series of 28 patients (20 male, mean age 66 years) were treated between January 2001 and July 2007. Sixteen patients had TAAAs type I III, one type IV, four thoracoabdominal placque ruptures, and seven patients CEAD. Patients were treated for asymptomatic, symptomatic, and ruptured aortic pathologies in 20, and 4 patients, respectively. Two patients had Marfan's syndrome; 61% had previous infrarenal aortic surgery. The infrarenal aorta was the distal landing zone in 70%. In elective cases, simultaneous approach (n = 9, group I) and staged approach (n = 11, group II) were performed. Mean follow-up is 22 months (range 0.1-78). RESULTS: Primary technical success was achieved in 89%. All stent grafts were implanted in the entire thoracoabdominal aorta. Additionally, three patients had previous complete arch vessel revascularization. Left subclavian artery was intentionally covered in three patients (11%). Thirty day mortality rate was 14.3% (4/28). One patient had a rupture before the staged endovascular procedure and died. Overall survival rate at 3 years was 70%, in group I 80%, and in group II 60% (P = .234). Type I endoleak rate was 8%. Permanent paraplegia rate was 11%. Three patients required long-term dialysis (11%). Peripheral graft occlusion rate was 11% at 30 days. Gut infarction with consecutive bowel resection occurred in two patients. There was no significant difference between group I and II regarding paraplegia and complications. CONCLUSIONS: Early results of visceral hybrid repair for high-risk patients with complex and extended TAAAs and CEADs are encouraging in a selected group of high risk patients in whom open repair is hazardous and branched endografts are not yet optional. PMID- 18381134 TI - Lessons learned from midterm follow-up of endovascular repair for traumatic rupture of the aortic isthmus. AB - OBJECTIVE: The aim of this study was to evaluate the short- and midterm results following endovascular repair of a traumatic rupture of the aortic isthmus. METHODS: Between January 2001 and January 2007, 27 patients underwent endovascular repair for acute traumatic rupture of the aortic isthmus (8 women, 19 men, mean age 40.2 +/- 16.7 years [19-78]). All patients underwent a computed tomography scan resulting in the preoperative diagnosis of aortic disruptions. Twenty-one patients were treated within the first 5 days following diagnosis. Follow-up computed tomography scans were performed at 1 week, at 3 and 6 months, and annually thereafter. The median follow-up was 40 months. RESULTS: All endografts were successfully deployed (Excluder-TAG [16], Talent [10], Zenith [2]). Three patients required common iliac artery access. The morbidity rate was 14.8%: two cases of inadvertent coverage of supra-aortic trunks occurred peroperatively, a proximal type I endoleak was successfully treated by a proximal implantation of a second endograft, and one collapse of an endograft was successfully treated by open repair and explantation. No patient suffered transient or permanent paraplegia, cerebral complication, endograft migration, or secondary endoleak. The overall mortality rate was 3.7%. CONCLUSIONS: Short and midterm results following endovascular treatment for traumatic rupture of the aortic isthmus favor the proposition of endovascular repair as the first-line treatment in hemodynamically unstable patients. In hemodynamically stable patients, the preoperative morphological evaluations aim to assess aortic anatomy and thereby detect possible technical limitations (aortic diameter <20 mm, severe aortic isthmus angulation, short proximal aortic neck <20 mm, conical aorta). In the presence of any one of these technical restrictions, open surgical treatment should be discussed to avoid major per- or postoperative complications related to endovascular repair. Further studies and long-term survival studies are mandatory to determine the efficacy and durability of this technique. PMID- 18381138 TI - Statins are independently associated with reduced mortality in patients undergoing infrainguinal bypass graft surgery for critical limb ischemia. AB - OBJECTIVE: Evidence suggesting a beneficial effect of cardioprotective medications in patients with lower extremity atherosclerosis derives largely from secondary prevention studies of heterogeneous populations. Patients with critical limb ischemia (CLI) have a large atherosclerotic burden with related high mortality. The effect of such therapies in this population is largely inferred and unproven. METHODS: The Project of Ex-Vivo vein graft Engineering via Transfection III (PREVENT III) cohort comprised 1404 patients with CLI who underwent lower extremity bypass grafting in a multicenter, randomized prospective trial testing the efficacy of edifoligide for the prevention of graft failure. Propensity scores were used to evaluate the influence of statins, beta blockers, and antiplatelet agents on outcomes while adjusting for demographics, comorbidities, medications, and surgical variables that may influence drug use. Primary outcomes were major adverse cardiovascular events < or =30 days, vein graft patency, and 1-year survival assessed by Kaplan-Meier method. Potential determinants of 1-year survival were modeled using a multivariate Cox regression. RESULTS: In this cohort, 636 patients (45%) were taking statins, 835 (59%) were taking beta-blockers, and 1121 (80%) were taking antiplatelet drugs. Perioperative major adverse cardiovascular events (7.8%) and early mortality (2.7%) were not measurably affected by the use of any drug class. Statin use was associated with a significant survival advantage at 1 year of 86% vs 81% (hazard ratio [HR], 0.71; 95% confidence interval [CI], 0.52-0.98; P = .03) by analysis of both unweighted and propensity score-weighted data. Use of beta-blockers and antiplatelet drugs had no appreciable impact on survival. None of the drug classes were associated with graft patency measures at 1 year. Significant predictors of 1-year mortality by Cox regression modeling were statin use (HR, 0.67; 95% CI, 0.51-0.90; P = .001), age >75 (HR, 2.1; 95% CI, 1.60-2.82; P = .001), coronary artery disease (HR, 1.5; 95% CI, 1.15-2.01; P = .001), chronic kidney disease stages 4 (HR, 2.0; 95% CI, 1.17-3.55; P = .001) and 5 (HR, 3.4; 95% CI, 2.39-4.73; P < .001), and tissue loss (HR, 1.9; 95% CI, 1.23-2.80; P = .003). CONCLUSIONS: Statin use is associated with improved survival in CLI patients 1 year after surgical revascularization. Further studies are indicated to determine optimal dosing in this population and to definitively address the question of relationship to graft patency. These data add to the growing literature supporting statin use in patients with advanced peripheral arterial disease. PMID- 18381141 TI - A prospective comparison of bilateral photoplethysmography versus the ankle brachial pressure index for detecting and quantifying lower limb peripheral arterial disease. AB - OBJECTIVE: This study prospectively assessed the diagnostic accuracy of a novel bilateral photoplethysmography toe pulse measurement technique for the detection of significant lower limb peripheral arterial disease. METHOD: Bilateral photoplethysmography toe pulse measurements were compared with the ankle-brachial pressure index (ABPI) gold standard reference. Pulse wave analysis techniques extracted timing, amplitude, and shape characteristics for the great toes and their right-to-left side differences. These characteristics were compared with previously obtained normative ranges, and the accuracy was assessed for all significant disease (ABPI <0.9) and higher-grade disease (ABPI <0.5). Measurements were collected in a controlled environment within a tertiary vascular surgical unit for 111 subjects (age range, 42-91 years), of whom 48 had significant lower limb peripheral arterial disease and 63 were healthy. Subjects were matched in age, sex, height, body mass index, and heart rate. Diagnostic performance was assessed using diagnostic sensitivity, specificity, accuracy, negative-predictive and positive-predictive value, and the kappa statistic representing agreement between techniques beyond chance. RESULTS: The degree that pulse shape fell beyond the normal range of normalized pulse shapes was at the threshold of substantial to almost perfect agreement compared with ABPI for significant disease detection (diagnostic accuracy, 91% [kappa = 0.80]; sensitivity, 93%; specificity, 89%), and with 90% accuracy (kappa = 0.65) for higher-grade disease detection. Pulse transit time differences between right and left toes also had substantial agreement with ABPI, with diagnostic accuracy of 86% for significant disease detection (pulse transit time to pulse foot [kappa = 0.71] and to pulse peak [kappa = 0.70]) and reached at least 90% for these for the higher-grade disease. The performance ranking for the different pulse features mirrored an earlier pilot study. With the shape and pulse transit time measurements, the negative-predictive values of the 5% disease population screening-prevalence level were at least 99% and had positive-predictive values of at least 98% for the 90% disease-prevalence level for vascular laboratory referrals. CONCLUSION: This simple-to-use technique could offer significant benefits for the diagnosis of peripheral arterial disease in settings such as primary care where noninvasive, accurate, and diagnostic techniques not requiring specialist training are desirable. Improved diagnosis and screening for peripheral arterial disease has the potential to allow identification and risk factor management for this high-risk group. PMID- 18381145 TI - Tear size and location impacts false lumen pressure in an ex vivo model of chronic type B aortic dissection. AB - BACKGROUND: Follow-up mortality is high in patients with type B aortic dissection (TB-AD) approaching one in four patients at 3 years. A predictor of increased mortality is partial thrombosis of the false lumen which may occlude distal tears. The hemodynamic consequences of differing tear size, location, and patency within the false lumen is largely unknown. We examined the impact of intimal tear size, tear number, and location on false lumen pressure. METHODS: In an ex-vivo model of chronic type B aortic dissection connected to a pulsatile pump, simultaneous pressures were measured within the true and false lumen. Experiments were performed in different dissection models with tear sizes of 6.4 mm and 3.2 mm in the following configurations; model A: proximal and distal tear simulating the most common hemodynamic state in patients with TB-AD; model B: proximal tear only simulating patients with partial thrombosis and occlusion of distal tear; and model C: distal tear only simulating patients sealed proximally via a stent graft with persistent distal communication. To compare false lumen diastolic pressure between models, a false lumen pressure index (FPI%) was calculated for all simulations as FPI% = (false lumen diastolic pressure/true lumen diastolic pressure) x 100. RESULTS: In model A, the systolic pressure was slightly lower in the false lumen compared with the true lumen while the diastolic pressure (DP) was slightly higher in the false lumen (DP 66.45 +/- 0.16 mm Hg vs 66.20 +/- 0.12 mm Hg, P < .001, FPI% = 100.4%). In the absence of a distal tear (model B), diastolic pressure was elevated within the false lumen compared with the true lumen (58.95 +/- 0.10 vs 54.66 +/- 0.17, P < .001, FPI% = 107.9%). The absence of a proximal tear in the presence of a distal tear (model C) diastolic pressure was also elevated within the false lumen versus the true lumen (58.72 +/- 0.24 vs 56.15 +/- 0.16, P < .001, FPI% 104.6%). The difference in diastolic pressure was greatest with a smaller tear (3.2 mm) in model B. In model B, DBP increased by 13.9% (P < .001, R(2) 0.69) per 10 beat per minute increase in heart rate (P < .001) independent of systolic pressure. CONCLUSIONS: In this model of chronic type B aortic dissection, diastolic false lumen pressure was the highest in the setting of smaller proximal tear size and the lack of a distal tear. These determinants of inflow and outflow may impact false lumen expansion and rupture during the follow-up period. PMID- 18381146 TI - Visualizing type IV endoleak using magnetic resonance imaging with a blood pool contrast agent. AB - Growing evidence suggests that graft porosity hampers aneurysm shrinkage in patients who have been treated with the original Excluder device. To our knowledge, this suspected porosity has never been visualized in such patients. We present three patients treated with the original Excluder device whose aneurysms did not shrink in the first 2 years after treatment. Computed tomography (CT) angiography and late phase CT did not show endoleak. We performed late phase magnetic resonance imaging with a blood pool agent to visualize graft porosity. Our cases illustrate the usability of a new contrast agent and a new imaging strategy for visualizing slow-flow endoleaks that can not be imaged using currently used imaging techniques with conventional contrast agents. PMID- 18381147 TI - Descending thoracic and abdominal aortic coarctation in the young: Surgical treatment after percutaneous approaches failure. AB - Descending thoracic and abdominal aortic coarctations are characterized by a segmental narrowing that frequently involves the origin of the visceral and renal arteries. Optimal primary treatment is debated, being reported for both surgical and percutaneous complications. We describe our surgical experience with two youths presenting with failure of distal descending aortic stenting and with abdominal aortic coarctation post-balloon angioplasty, and associated thrombosis of a stented right renal artery and stenosis of the origin of the superior mesenteric artery (SMA). In both cases, a longitudinal aortoplasty was performed with a polytetrafluoroethylene (PTFE) patch, using simple aortic cross-clamping. Renal thrombosis and SMA stenosis were managed with eversion technique. In hospital course was uneventful. Midterm follow-up showed absence of significant restenosis and better control of hypertension. In order to refrain from operating on these patients as long as possible, and also because of the very high risk of a redo-surgery, we think that an initial balloon angioplasty should be considered. Surgical management can be adopted, even after failure of percutaneous treatments, with satisfactory short- and midterm vessels patency. PMID- 18381148 TI - Thoracic aorta endograft as an adjunct to resection of a locally invasive tumor: a new indication to endograft. AB - Covered stent grafts are currently used for arterial aneurysm exclusion, aortic dissection, or peripheral occlusive disease. A new indication to endograft was applied to perform resection of the thoracic aorta for infiltration of an adjacent lung cancer into the vessel wall, avoiding a major vascular intervention for aortic graft interposition associated with tumor resection. PMID- 18381149 TI - Bilateral renal artery aneurysm rupture in a man with leukemia: report of a case. AB - Renal artery aneurysms, once thought to be rare, are diagnosed more frequently due to the increasing use of computed tomography, angiography, and other imaging to delineate pathology. The incidence is less than 1% in the general population,(1,2) and increases to 2.5% in the hypertensive population undergoing angiography.(3) Incidence approaches 10% in autopsy series.(4) Despite increasing incidence, renal artery aneurysm rupture remains uncommon. We report the case of a man with chronic myelomonocytic leukemia who suffered bilateral renal artery aneurysm rupture over the course of 1 month. PMID- 18381150 TI - A giant aneurysm of a branch of axillary artery. PMID- 18381151 TI - Basic research by vascular surgeons: why it is important and how to keep it going. PMID- 18381152 TI - Regarding "transcervical carotid stenting with carotid artery flow reversal: 3 year follow-up of 103 stents". PMID- 18381154 TI - Regarding a comparison of percutaneous transluminal angioplasty versus amputation for critical limb ischemia in patients unsuitable for open surgery. PMID- 18381156 TI - Regarding bare metal stent infections: case report and review of the literature. PMID- 18381158 TI - Regarding endoleaks after endovascular aneurysm repair lead to nonuniform intra aneurysm sac pressure. PMID- 18381159 TI - Regarding "does simvastatin save lives; If so, when and in whom?". PMID- 18381161 TI - Regarding "light assisted stab phlebectomy: report of a technique for removal of lower extremity varicose veins". PMID- 18381163 TI - Natural orifice translumenal endoscopic surgery. Foreword. PMID- 18381164 TI - Natural orifice translumenal endoscopic surgery. Preface. PMID- 18381165 TI - Evolution of laparoscopic surgery: lessons for NOTES. AB - Natural orifice translumenal endoscopic surgery (NOTES) is considered by some to be the next revolution in surgery. To advance into clinical NOTES safely and responsibly, the lessons learned from laparoscopic surgery should be applied to the development of NOTES as much as possible. This novel technique is still in evolution and newly designed instruments and devices should be developed. Industry has a considerable role in research and development and collaboration between clinicians and engineers for this matter is crucial. PMID- 18381166 TI - Techniques for transgastric access to the peritoneal cavity. AB - Natural orifice translumenal endoscopic surgery (NOTES) is a unique emerging surgical concept expanding flexible endoscopy beyond the gut wall. The methods and technology growing from this concept may minimize trauma from surgical access to the peritoneal cavity by completely eliminating body surface incisions. So far, NOTES surgeries have been reported by modifying laparoscopic surgery. The peroral transgastric route was chosen to access the peritoneal cavity in initial trials because of a potentially lower risk for surrounding organ injury using the anterior wall percutaneous endoscopic gastrostomy-style gastrotomy. This article reviews and describes techniques of transgastric access to the peritoneal cavity used in published animal studies. PMID- 18381167 TI - Techniques for transanal access to the peritoneal cavity. AB - Natural orifice translumenal endoscopic surgery (NOTES) represents a burgeoning but still largely experimental field. For surgeries involving the upper abdominal organs, the transanal approach promises to provide a more direct route in contrast to the often cumbersome retroflexion typically required with the transgastric approach. The potential disadvantages of the transanal route are also significant and include issues of sterility, the risk of inadvertent trauma to adjacent organs during transmural puncture, and the risk of colonic wall shearing. This article reviews the evolution of transanal access to the peritoneal cavity, highlights the various techniques that have been used for transanal access and closure, and discusses in further detail the relative advantages and disadvantages of this approach. PMID- 18381168 TI - NOTES and anastomosis. AB - The history and development for forming anastomoses using natural orifice translumenal endoscopic surgery (NOTES) techniques is described. Sutured gastrojejunostomy, enteroenteral anastomosis, and ileocolonic anastomoses using rigid and circular staplers passed through the rectum and vaginal wall using a transgastric supervising endoscope using a natural orifice translumenal endoscopic surgery hybrid approach. The staplers for this type of approach have to change. They need to be longer, more flexible, and able to change shape (eg, becoming smaller for introduction and then expanding in the peritoneal cavity). Different methods of introduction over guidewires or in combination with flexible gastroscopes are also needed. There is still a requirement for more ingenuity, persistence, and research if the goal of safer effective formation of anastomoses by less invasive means is to be realized. PMID- 18381169 TI - The eagle or the snake: platforms for NOTES and radical endoscopic therapy. AB - It has only been about 3 years since natural orifice translumenal endoscopic surgery (NOTES) first began to gather the attention of the medical and surgical community at large. The concepts behind NOTES, however, have been under development for almost a decade. It is important to revisit some basic concepts regarding therapeutic flexible endoscopy and in the process understand the fundamental premises on which a sound technology development program for NOTES should be built. This article provides such a review and emphasizes general principles rather than specific embodiments, which are very much at a nascent stage. PMID- 18381170 TI - Infection prevention in NOTES. AB - Prevention of infection during natural orifice translumenal endoscopic surgery (NOTES) was identified as one of the most important challenges for translumenal surgery. Does infection prevention during NOTES warrant such attention? This article summarizes the accumulated data about septic complications during translumenal surgery. PMID- 18381171 TI - EUS-guided NOTES interventions. AB - Entering the peritoneal cavity with the echoendoscope has been avoided because this endoscope is rather rigid and difficult to handle and maneuver in a limited space. Endoscopic ultrasound may be of help, however, to guide natural orifice translumenal endoscopic surgery (NOTES) procedures. This article provides an overview of the potential benefit and the few endoscopic ultrasound-based natural NOTES procedures performed to date. PMID- 18381172 TI - Spatial orientation and off-axis challenges for NOTES. AB - Issues of spatial orientation and off-axis visualization must be addressed to make natural orifice translumenal endoscopic surgery (NOTES) more than a clinical oddity practiced by a highly skilled minority. The issue is the degree of mental work needed to operate with this new modality, with issues of depth perception and anatomy recognition currently making these procedures extremely difficult and unlikely to be practically applied or taught. New instrument designs and clinical tricks combined can dramatically decrease the mental work needed and make this closer to a widely learnable access method. In the long run, computer interfaces may be necessary to resolve some of these human factor issues and make a true universal platform. PMID- 18381173 TI - Hybrid surgery: combined laparoscopy and natural orifice surgery. AB - Hybrid procedures combine laparoscopy and natural orifice surgery techniques. Some groups are currently using hybrid procedures to investigate the concepts of pure natural orifice surgery. Thus far, hybrid procedures have validated the safety of transgastric peritoneoscopy, although improved instrumentation is imperative before widespread clinical applications. As technical improvements develop from hybrid surgery, gastrointestinal endoscopy and abdominal surgery could be revolutionized. PMID- 18381174 TI - Transition from laboratory to clinical practice in NOTES: role of NOSCAR. AB - The way to move natural orifice translumenal endoscopic surgery (NOTES) from the laboratory to the clinical arena is to do high-quality research in the areas recommended by the NOSCAR working group representing barriers to NOTES procedures. Additionally, good experimental work needs to be done to determine whether the theorized advantages of the NOTES approach actually represent reality. It is anticipated that monies will continue to flow to sponsor the necessary research in NOTES. It is clear that momentum continues to build in this exciting new potential paradigm shift in minimally invasive surgery, but one of the keys is to produce good science to validate the NOTES approach. PMID- 18381175 TI - Training for NOTES. AB - The foundation of skills for the performance of natural orifice translumenal endoscopic surgery (NOTES) lies in the training for general surgery (especially laparoscopy) and flexible gastrointestinal endoscopy. Physicians wishing to practice NOTES need to acquire or have both skill sets, or need to partner together to blend complementary capabilities with colleagues. In the future, however, a new cadre of NOTES specialists may emerge who will have developed individual expertise in the full spectrum of NOTES knowledge base requirements. This article highlights a body of knowledge and skills needed to become a NOTES proceduralist and review the current training paradigms for gastrointestinal endoscopists and surgeons. PMID- 18381176 TI - NOTES: human experience. AB - Human natural orifice translumenal endoscopic surgery (NOTES) is already being reported from numerous centers, and the results seem promising. There are key issues to be addressed and benefits over traditional safe procedures need to be demonstrated. Interestingly, however, human NOTES seem to be progressing at a fast pace compared with the evolution of surgical procedures or techniques described previously. This article examines the current status of NOTES in humans, the challenges, and the implications on the future of minimal access surgery. PMID- 18381177 TI - Designing clinical trials for NOTES. AB - This article breaks down the elements of designing clinical trials, with an emphasis on randomized controlled trials, as applied to advancing natural orifice translumenal endoscopic surgery research. It discusses research ethics, feasibility, design elements (such as randomization, blinding, and choosing a primary outcome), and issues regarding presentation of results. The pros and cons of subjective quality of life outcomes are debated, and the role of the patient/consumer in driving the direction and interpretation of clinical trials is discussed. PMID- 18381178 TI - NOTES: the future. AB - The concept of natural orifice translumenal endoscopic surgery (NOTES) has grown in acceptance since the time of its introduction in 2000. Developments in techniques of peritoneal access and closure, surgical techniques, and equipment modification have already been published and intensive research is ongoing. Current and future endoscopists will reap the benefit of this research because many techniques and devices that are developed for NOTES will enhance the ability to perform luminal intervention, including polypectomy, endoluminal hemostasis, and submucosal resection. The authors attempt to predict the future of NOTES by describing potential applications for certain clinical scenarios and conditions. PMID- 18381179 TI - Development and preliminary psychometric testing of a new OA pain measure--an OARSI/OMERACT initiative. AB - OBJECTIVE: To evaluate the measurement properties of a new osteoarthritis (OA) pain measure. METHODS: The new tool, comprised of 12 questions on constant vs intermittent pain was administered by phone to 100 subjects aged 40+ years with hip or knee OA, followed by three global hip/knee questions, the Western Ontario and McMaster Universities (WOMAC) pain subscale, the symptom subscales of the Hip Disability and OA Outcome Score (HOOS) or Knee Injury and OA Outcome Score (KOOS), and the limitation dimension of the Late Life Function and Disability Instrument (LLFDI). Test-retest reliability was assessed by re-administration after 48-96h. Item response distributions, inter-item correlations, item-total correlations and Cronbach's alpha were assessed. Principle component analysis was performed and test-retest reliability was assessed by intra-class correlation coefficient (ICC). RESULTS: There was good distribution of response options across all items. The mean intensity was higher for intermittent vs constant pain, indicating subjects could distinguish the two concepts. Inter-item correlations ranged from 0.37 to 0.76 indicating no item redundancy. One item, predictability of pain, was removed from subsequent analyses as correlations with other items and item-total correlations were low. The 11-item scale had a corrected inter-item correlation range of 0.54-0.81 with Cronbach's alpha of 0.93 for the combined sample. Principle components analysis demonstrated factorial complexity. As such, scoring was based on the summing of individual items. Test retest reliability was excellent (ICC 0.85). The measure was significantly correlated with each of the other measures [Spearman correlations -0.60 (KOOS symptoms) to 0.81 (WOMAC pain scale)], except the LLFDI, where correlations were low. CONCLUSIONS: Preliminary psychometric testing suggests this OA pain measure is reliable and valid. PMID- 18381180 TI - Safety and tolerability of angiotensin-converting enzyme inhibitor versus the combination of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker in patients with left ventricular dysfunction: a systematic review and meta-analysis of randomized controlled trials. AB - BACKGROUND: The addition of an angiotensin receptor blocker (ARB) to an angiotensin-converting enzyme inhibitor (ACEI) in patients with heart failure remains controversial. A recent meta-analysis showed that the combination therapy reduces hospitalization without improved survival. Whether excess risk is associated with this strategy has not been fully explored. We sought to quantify the risk of adverse events of combination therapy (ACEI+ARB) versus ACEI alone. METHODS: MEDLINE, EMBASE, BIOSIS, and Cochrane databases were searched. Eligible studies were randomized, placebo-controlled trials of ACEI versus the combination of ACEI+ARB in patients with heart failure or left ventricular dysfunction. Included studies were reviewed to determine the frequency of adverse effects leading to discontinuation of therapy. RESULTS: Nine trials that enrolled 18,160 patients met the inclusion criteria. A total of 9199 patients received combination therapy, and 8961 patients received an ACEI only. Patients receiving combination therapy had an increased risk of developing any adverse effect by 2.3% (relative risk [RR] = 1.27, 95% confidence interval [CI] = 1.15-1.40, P < .00001, I(2) = 15.9%, number needed to harm [NNH] = 42), hypotension by 1.1% (RR = 1.91, 95% CI = 1.37-2.66, P = .0002, I(2) = 26.6%, NNH = 89), worsening renal function by 1% (RR = 2.12, 95% CI = 1.30-3.46, P = .003, I(2) = 67.3%, NNH = 100), and hyperkalemia by 0.6% (RR = 4.17, 95% CI = 2.31-7.53, P < .00001, I(2) = 0%, NNH = 149). There was no difference in angioedema (RR = 0.88, 95% CI = 0.43 1.80, P = .72, I(2) = 0%) or cough (RR = 0.84, 95% CI = 0.65-1.09, P = .19, I(2) = 0%). CONCLUSION: The current cumulative evidence suggests that patients with left ventricular dysfunction have an increased risk of adverse events leading to discontinuation on ACEI+ARB combination therapy compared with ACEI alone. This excess risk, coupled with a lack of consistent mortality benefit, suggests that ARBs should not routinely be added to ACEI therapy for left ventricular dysfunction. If chosen, the combination strategy may warrant closer patient monitoring to detect adverse effects. PMID- 18381181 TI - Effects of 5'-phosphodiesterase four-week long inhibition with sildenafil in patients with chronic heart failure: a double-blind, placebo-controlled clinical trial. AB - BACKGROUND: The effects of chronic inhibition of 5'-phosphodiesterase with sildenafil on functional capacity, ventilatory efficiency, oxygen uptake, pulmonary hypertension, and endothelial function in chronic heart failure (CHF) are unknown. METHODS: We conducted a randomized, double-blind, placebo-controlled trial to assess the acute (1 hour after 50 mg by mouth) and chronic (4 weeks after 50 mg 3 times per day by mouth) effects of sildenafil in outpatients with CHF. The outcomes were cardiopulmonary exercise test parameters (chronic effect), echocardiographic-derived pulmonary artery systolic pressure, and plethysmography derived forearm blood flow (acute and chronic effects). RESULTS: Nineteen patients with CHF (48 +/- 12 years) with an ejection fraction of 28% +/- 6% were studied. Patients who received sildenafil (n = 11) showed improved maximal oxygen uptake, ventilatory efficiency, and oxygen uptake kinetics. Sildenafil decreased pulmonary artery systolic pressure levels at 60 minutes and at 4 weeks compared with changes after placebo (P = .004 for group and time interaction). Improvement in ventilatory efficiency was positively associated with reductions in pulmonary artery systolic pressure. Patients allocated to placebo demonstrated a trend toward decreased forearm blood flow after reactive hyperemia, whereas this remained unchanged in patients allocated to sildenafil. CONCLUSIONS: Sildenafil administration for 4 weeks in stable outpatients with CHF improves functional capacity, ventilatory efficiency, oxygen uptake kinetics, and pulmonary hypertension. These effects may be mediated in part by improvements in endothelial function. PMID- 18381182 TI - Improvement in dilated cardiomyopathy after bariatric surgery. AB - BACKGROUND: Young severely obese patients with advanced heart failure may not be suitable candidates for cardiac transplantation because of surgical morbidity and availability of adequately sized donor hearts. METHODS AND RESULTS: We report 2 patients with severe systolic dysfunction and Class IV heart failure despite maximal medical therapy who were considered for cardiac transplantation. Because of their severe obesity, transplantation was not considered an optimal therapy, and both were referred for bariatric surgery. The individuals had nonischemic cardiomyopathy. Both underwent laparoscopic vertical gastrectomy, minimizing surgical risk while providing definitive reduction in gastric volume. They experienced substantial weight loss and resolution of dyspnea. Inotrope infusion was discontinued in 1 dobutamine-dependent individual. They achieved weight reduction of 46 to 52 kg after the surgery. End-systolic volume index improved from 64 to 49 mL/m(2) and from 66 to 39 mL/m(2). Left ventricular ejection fraction improved from 20% to 45% and from 25% to 39%. They remain symptom-free and are no longer listed for transplant at 2 years' follow-up. CONCLUSIONS: Bariatric surgery may lead to improvement in left ventricular systolic dysfunction in young morbidly obese individuals with nonischemic cardiomyopathy. Potential explanations for the improvement in left ventricular function include reduced direct toxic effects of adiposity on cardiomyocytes and improved hemodynamics after weight loss. The potential for bariatric surgery to provide an alternative to heart transplantation in extreme obesity merits further study. PMID- 18381183 TI - Objectively measured, but not self-reported, medication adherence independently predicts event-free survival in patients with heart failure. AB - OBJECTIVE: Medication nonadherence is presumed to be related to poor clinical outcomes, yet this relationship rarely has been tested using objective adherence measures in patients with heart failure. Which objective indicators of medication adherence predict clinical outcomes are unknown. The study objective was to determine which indicators of medication adherence are predictors of event-free survival. METHODS: Patients (N = 134) with heart failure (69% were male, aged 61 +/- 11 years, 61% with New York Heart Association class III/IV heart disease) were enrolled in this 6-month longitudinal study. Adherence was measured using two measures: 1) an objective measure, the Medication Event Monitoring System (MEMS); and 2) self-reported adherence (Medical Outcomes Studies Specific Adherence Scale). Three indicators of adherence were assessed by MEMS: 1) dose count, percentage of prescribed doses taken; 2) dose-days, percentage of days correct number of doses taken; and 3) dose-time, percentage of doses taken on schedule. Events (emergency department visits, rehospitalization, and mortality) were obtained by patient/family interview and hospital databases. RESULTS: In Cox regression, two of the three MEMS indicators, dose-count and dose-day, predicted event-free survival before and after controlling for age, gender, ejection fraction, New York Heart Association class, angiotensin-converting enzyme inhibitor use, and beta-blocker use (P = .004, P = .008, and P = .224, respectively). Self-report adherence did not predict outcomes (P = .402). CONCLUSION: Dose-count and dose-day predicted event-free survival. Neither dose time nor self-reported adherence predicted outcomes. Health care providers should assess specific behaviors related to medication taking rather than a global patient self-assessment of patient adherence. PMID- 18381184 TI - Incident heart failure hospitalization and subsequent mortality in chronic heart failure: a propensity-matched study. AB - OBJECTIVE: Hospitalization for worsening heart failure (HF) is common and associated with high mortality. However, the effect of incident HF hospitalization (compared with no HF hospitalization) on subsequent mortality has not been studied in a propensity-matched population of chronic HF patients. METHODS: In the Digitalis Investigation Group trial, 5501 patients had no HF hospitalizations (4512 alive at 2 years after randomization) and 1732 patients had HF hospitalizations during the first 2 years (1091 alive at 2 years). Propensity scores for incident HF hospitalization during the first 2 years after randomization were calculated for each patient and used to match 1057 patients (97%) who had 2-year HF hospitalization with 1057 patients who had no HF hospitalization. We used matched Cox regression analysis to estimate the effect of incident HF hospitalization during the first 2 years after randomization on post-2-year mortality. RESULTS: Compared with 153 deaths (rate, 420/10,000 person years) in the no HF hospitalization group, 334 deaths (rate, 964/10,000 person years) occurred in the HF hospitalization group (hazard ratio 2.49; 95% confidence interval 1.97-3.13; P < .0001). The hazard ratios (95% confidence intervals) for cardiovascular and HF mortality were 2.88 (2.23-3.74; P < .0001) and 5.22 (3.34-8.15; P < .0001), respectively. CONCLUSIONS: Hospitalization for worsening HF was associated with increased risk of subsequent mortality in ambulatory patients with chronic HF. These results highlight the importance of HF hospitalization as a marker of disease progression and poor outcomes in chronic HF, reinforcing the need for prevention of HF hospitalizations and strategies to improve postdischarge outcomes. PMID- 18381185 TI - B-type natriuretic Peptide-guided treatment for predicting outcome in patients hospitalized in sub-intensive care unit with acute heart failure. AB - BACKGROUND: Studies with natriuretic peptides have suggested that physicians do not treat heart acute failure (AHF) aggressively enough, and predischarge B-type natriuretic peptide (BNP) levels may be a useful reminder that more treatment is required. The purpose of this study was to demonstrate that variations in BNP levels during hormone-guided treatment and measured body hydration status enable the timing of the patient's discharge to be optimized. METHODS AND RESULTS: We retrospectively evaluated 186 patients admitted for AHF. All subjects underwent serial bioelectrical impedance analysis and BNP measurement. Therapy was titrated according to BNP value to reach a BNP value of <250 pg/mL, whenever is possible. A BNP value on discharge of <250 pg/mL (obtained in 54% of the patients) predicted a 16% event rate within 6 months, whereas a value >250 pg/mL was associated with a far higher percentage (78%) of adverse events. Among the former, no significant differences in event rate were seen in relation to the time necessary to obtain a reduction in BNP values below 250 pg/mL (14 versus 18%, chi-square = 0.3, NS). Cox regression showed that a BNP cutoff value of 250 pg/mL is the most accurate predictors of events. CONCLUSIONS: Our study demonstrates the usefulness of BNP in intrahospital stratification of AHF, in the decision-making process, and as a tool for "tailored therapy." Integrating this approach into the routine assessment of HF patients would allow clinicians to more accurately identify high-risk patients, who may derive increased benefit from intensive in-hospital management strategies. PMID- 18381186 TI - Predischarge QRS score and risk for heart failure after first ST-elevation myocardial infarction. AB - BACKGROUND: The prognostic value of the QRS score, a simple index of infarct size after a first ST-elevation myocardial infarction, has not been adequately explored in the reperfusion era. METHODS AND RESULTS: We prospectively followed up 100 consecutive survivors of a first ST-elevation myocardial infarction (aged 64 +/- 13 years, 77% were male) without bundle branch block or paced rhythm at hospital discharge for 3 months. The modified 32-point QRS score was calculated as part of the predischarge evaluation. The predefined primary endpoint was the composite of death or hospitalization for heart failure. By 3 months, 6 patients died and 16 patients were readmitted for heart failure, resulting in a 22% primary endpoint rate. Patients with a QRS score >/= 3 at hospital discharge (n = 38) had significantly more events compared with those with a QRS score < 3 (44.7% vs. 8.2%, P < .001), and all six deaths occurred among patients with a QRS score >/= 3 (P = .002). A QRS score < 3 reliably predicted heart-failure free survival during the follow-up period (negative predictive value 91.9%). In multivariate models, the QRS score was an independent predictor of the primary endpoint (hazard ratio = 1.4 per point, 95% confidence interval 1.1-1.8, P = .003). CONCLUSION: For patients surviving a first ST-elevation myocardial infarction, the predischarge QRS score provides powerful prognostic information on short-term outcomes, including mortality and readmission for heart failure. PMID- 18381187 TI - Management of atrial fibrillation in patients with heart failure. AB - BACKGROUND: There is a well-documented relationship and a complex interaction between atrial fibrillation (AF) and heart failure. The coexistence of these 2 clinical entities renders their management even more challenging. METHODS AND RESULTS: We searched current literature to review the management of AF in patients with heart failure. The cornerstones of AF treatment are rate control, cardioversion, and maintenance of sinus rhythm (SR), and prevention of thromboembolism. The issue of rhythm versus rate control remains unresolved. Nonpharmacologic therapies such as radiofrequency catheter ablation of the atrioventricular node with permanent pacemaker implantation, curative catheter ablation of AF, and cardiac resynchronization therapy are emerging and may alter the management of these patients. CONCLUSION: Treatment of atrial fibrillation in the setting of heart failure encompasses a variety of approaches including drugs, devices, and ablation. Larger randomized trials are required to clarify the management of such patients. PMID- 18381188 TI - Xanthine oxidase inhibition improves left ventricular dysfunction in dilated cardiomyopathic hamsters. AB - BACKGROUND: Oxidative stress is implicated in cardiac remodeling and failure. We tested whether xanthine oxidase (XO) inhibition could decrease myocardial oxidative stress and attenuate left ventricular (LV) remodeling and dysfunction in the TO-2 hamster model of dilated cardiomyopathy. METHODS AND RESULTS: TO-2 hamsters were randomized to treatment with the XO inhibitor, allopurinol, or vehicle from 6 to 12 weeks of age. F1B hamsters served as controls. TO-2 hamsters treated with vehicle progressively developed severe LV systolic dysfunction and dilation between 6 and 12 weeks. Marked cardiac fibrosis was apparent in these hamsters at 12 weeks in comparison with F1B controls. The ratio of reduced to oxidized glutathione (GSH/GSSG) was decreased and malondialdehyde levels were increased in the hearts of vehicle-treated TO-2 hamsters. Treatment with allopurinol from 6 to 12 weeks attenuated LV dysfunction and dilation as well as myocardial fibrosis and the upregulation of a fetal-type cardiac gene. Allopurinol also inhibited both the decrease in GSH/GSSG ratio and the increase in malondialdehyde levels in the heart. CONCLUSIONS: These results indicate that chronic XO inhibition with allopurinol attenuates LV remodeling and dysfunction as well as myocardial oxidative stress in this model of heart failure. Allopurinol may prove beneficial for the treatment of heart failure. PMID- 18381189 TI - 17 Beta-estradiol differentially affects left ventricular and cardiomyocyte hypertrophy following myocardial infarction and pressure overload. AB - BACKGROUND: We have shown previously that 17beta-estradiol (E2) increases left ventricular (LV) and cardiomyocyte hypertrophy after myocardial infarction (MI). However, E2 decreases hypertrophy in pressure overload models. We hypothesized that the effect of estrogen on cardiac hypertrophy was dependent on the type of hypertrophic stimulus. METHODS AND RESULTS: Ovariectomized wild-type female mice (n = 192) were given vehicle or E2 treatment followed by coronary ligation (MI), transverse aortic constriction (TAC), or sham operation. Signaling pathway activation was studied at 3, 24, and 48 hours, whereas echocardiography and hemodynamic studies were performed at 14 days. MI induced early but transient activation of p38 and p42/44 MAPK pathways, whereas TAC induced sustained activation of both pathways. E2 had no effect on these pathways, but increased Stat3 activation after MI while decreasing Stat3 activation after TAC. MI caused LV dilation and decreased fractional shortening (FS) that were unaltered by E2. TAC caused LV dilation, reduced FS, and increased LV mass, but in this model, E2 improved these parameters. After MI, E2 led to increases in myocyte cross sectional area, atrial natriuretic peptide (ANP) and beta-myosin heavy chain (MHC) gene expression, but E2 diminished TAC-induced increases ANP and beta-MHC gene expression. CONCLUSIONS: These data demonstrate that the effects of E2 on LV and myocyte remodeling depend on the nature of the hypertrophic stimulus. The opposing influence of E2 on hypertrophy in these models may, in part, result from differential effects of E2 on Stat3 activation. Further work will be necessary to explore this and other potential mechanisms by which estrogen affects hypertrophy in these models. PMID- 18381191 TI - Are ultrasound lung comets useful as prognostic indicator in any patients with dyspnea or chest pain? PMID- 18381190 TI - Simultaneous right atrioventricular pacing: a novel model to study atrial remodeling and fibrillation in the setting of heart failure. AB - BACKGROUND: Atrial fibrillation (AF) is a common arrhythmia which contributes to morbidity and mortality in patients with heart failure (HF). Atrial remodeling is a key substrate for the development of AF in HF. However, experimental models that study AF in the setting of HF have important limitations. We evaluated a new dog model of atrial remodeling and AF. METHODS AND RESULTS: Twenty-two mongrel dogs were randomized into 2 groups: 14 dogs with simultaneous atrioventricular pacing (SAVP) for 2 weeks (220 beats/min, no AV delay) and 8 control dogs with no pacing. SAVP for 2 weeks induced marked changes in atrial mechanical function and conduction. Left atrial area fractional shortening decreased 61 +/- 17%, whereas left ventricular area fractional shortening decreased by 38 +/- 18% from baseline (both P < .05). Conduction slowed and conduction heterogeneity increased. AF was induced in 83% of SAVP dogs, lasting a median of 1600 seconds, versus no dogs with induced AF in the controls. SAVP significantly increased nonfibrillar collagen in the mid-myocardium of both atrial appendages and matrix metalloproteinase-9 activity. CONCLUSIONS: SAVP in dogs induces structural and electrical remodelling that form the substrate for reproducibly inducible AF. This novel model may be useful for studies of the pathophysiology and treatment of AF in heart failure. PMID- 18381193 TI - High-performance liquid chromatography accelerator mass spectrometry: correcting for losses during analysis by internal standardization. AB - A method was developed to account for analytical losses of (14)C-analyte when determining the concentration in biological samples using chromatographic separation and analysis by accelerator mass spectrometry. From the equations of J. Vogel and A.H. Love (in: A.L. Burlingame (Ed.), Methods in Enzymology, Academic Press, New York, 2005), new equations were derived to describe the isotopic dilution of a chromatographically isolated (14)C-analyte. The analytical recovery for each sample was determined by the use of the UV response for nonlabeled analyte, as an internal standard against a standard curve. The slope of the curve was substituted into the equations to provide a method of accurately determining the analyte concentration. PMID- 18381194 TI - Surface plasmon resonance-enhanced fluorescence implementation of a single-step competition assay: demonstration of fatty acid measurement using an anti-fatty acid monoclonal antibody and a Cy5-labeled fatty acid. AB - The development of a single-step, separation-free method for measurement of low concentrations of fatty acid using a surface plasmon resonance-enhanced fluorescence competition assay with a surface-bound antibody is described. The assay behavior was unexpectedly complex. A nonlinear coverage-dependent self quenching of emission from surface-bound fluorescent label was deduced from the response kinetics and attributed to a surface plasmon-mediated energy transfer between adsorbed fluorophores, modified by the effects of plasmon interference. Principles of assay design to avoid complications from such effects are discussed. An anti-fatty acid mouse monoclonal antibody reacting to the alkyl chain was prepared and supported on a gold chip at a spacing appropriate for surface-plasmon field-enhanced fluorescence spectroscopy (SPEFS), by applying successively a self-assembled biotinylated monolayer, then streptavidin, then biotinylated protein A, and then the antibody, which was crosslinked to the protein A. Synthesis of a fluorescently (Cy5) tagged C-11 fatty acid is reported. SPEFS was used to follow the kinetics of the binding of the labeled fatty acid to the antibody, and to implement a competition assay with free fatty acid (undecanoic acid), sensitive at the 1 microM scale, a sensitivity limit caused by the low affinity of antibodies for free fatty acids, rather than the SPEFS technique itself. Free fatty acid concentration in human serum is in the range 0.1-1mM, suggesting that this measurement approach could be applied in a clinical diagnostic context. Finally, a predictive, theoretical model of fatty acid binding was developed that accounted for the observed "overshoot" kinetics. PMID- 18381195 TI - Development and optimization of a binding assay for histone deacetylase 4 using surface plasmon resonance. AB - Histone deacetylase 4 (HDAC4) is a histone deacetylase profoundly involved in cell differentiation and in the pathogenesis of cancer. The histone deacetylase inhibitors are a new, promising class of anticancer agents. The screening of molecular interactions involving determination of the affinity of drug candidates is an integral part of the drug discovery process. Here we report the development of an assay using surface plasmon resonance for the analysis of HDAC4-small molecule interactions. We describe a new cloning and purification strategy that can be used to set up surface plasmon resonance experiments with other recombinant proteins. PMID- 18381196 TI - Electrochemical biosensor for the detection of cauliflower mosaic virus 35 S gene sequences using lead sulfide nanoparticles as oligonucleotide labels. AB - Lead sulfide (PbS) nanoparticles were synthesized in aqueous solution and used as oligonucleotide labels for electrochemical detection of the 35 S promoter from cauliflower mosaic virus (CaMV) sequence. The PbS nanoparticles were modified with mercaptoacetic acid and could easily be linked with CaMV 35 S oligonucleotide probe. Target DNA sequences were covalently linked on a mercaptoacetic acid self-assembled gold electrode, and DNA hybridization of target DNA with probe DNA was completed on the electrode surface. PbS nanoparticles anchored on the hybrids were dissolved in the solution by oxidation of HNO3 and detected using a sensitive differential pulse anodic stripping voltammetric method. The detection results can be used to monitor the hybridization reaction. The CaMV 35 S target sequence was satisfactorily detected with the detection limit as 4.38 x 10(-12)mol/L (3sigma). The established method extends nanoparticle-labeled electrochemical DNA analysis to specific sequences from genetically modified organisms with higher sensitivity and selectivity. PMID- 18381197 TI - Tissue digestion for aluminum determination in experimental animal studies. AB - Four different procedures for the determination of aluminum in tissues by atomic absorption spectrometry (AAS) were investigated. They consisted of conventional acid digestion carried out before and after sample drying, associated or not with fat extraction. Drying was carried out in a conventional oven at 65 degrees C for 24 h. For fat extraction, different solvents and solvent mixtures were investigated considering both extraction yield and sample adequacy for further AAS measurement. Acid digestion was carried out with pure HNO3 or with its mixture with HClO4. After digestion, aluminum was measured by graphite furnace atomic absorption spectrometry. Tissues were collected from Al-exposed and nonexposed mice. The results indicated that drying the sample prior to digestion is advantageous as the amount of acid necessary can be significantly reduced. This procedure does not contribute to increase the aluminum level in the samples providing that careful measures to avoid contamination are taken, as the same procedures carried out without taking any precautions to avoid contamination produced imprecise results. Finally, aluminum was not found in the fatty fraction of any sample, even in exposed mice, demonstrating that aluminum does not accumulate in this part of the tissues. PMID- 18381198 TI - Blue native electrophoresis study on lipases. AB - We have developed a modified blue native polyacrylamide gel electrophoresis (PAGE) protocol that can overcome aggregation of lipases seen in native PAGE. We have shown that two lipases, Pseudomonas aeruginosa lipase and Candida rugosa lipase, which aggregate in the native gel, can be resolved using our protocol. Activity staining was done to test for the functionality of the two lipases. PMID- 18381199 TI - Characterization of Cu2+-binding modes in the prion protein by visible circular dichroism and multivariate curve resolution. AB - Visible circular dichroism (CD) spectra from the copper(II) titration of the metal-binding region of the prion protein, residues 57-98, were analyzed using the self-modeling curve resolution method multivariate curve resolution alternating least squares (MCR-ALS). MCR-ALS is a set of mathematical tools for estimating pure component spectra and composition profiles from mixture spectra. Model-free solutions (e.g., soft models) are produced under the assumption that pure component profiles should be nonnegative and unimodal. Optionally, equality constraints can be used when the concentration or spectrum of one or more species is known. MCR-ALS is well suited to complex biochemical systems such as the prion protein which binds multiple copper ions and thus gives rise to titration data consisting of several pure component spectra with overlapped or superimposed absorption bands. Our study reveals the number of binding modes used in the uptake of Cu(2+) by the full metal-binding region of the prion protein and their relative concentration profiles throughout the titration. The presence of a non CD active binding mode can also be inferred. We show that MCR-ALS analysis can be initialized using empirically generated or mathematically generated pure component spectra. The use of small model peptides allows us to correlate specific Cu(2+)-binding structures to the pure component spectra. PMID- 18381200 TI - Suppression of protein l-isoaspartyl (d-aspartyl) methyltransferase results in hyperactivation of EGF-stimulated MEK-ERK signaling in cultured mammalian cells. AB - l-Aspartyl (l-Asp) and l-asparaginyl residues in proteins isomerize or racemize to d,l-isoaspartyl (d,l-isoAsp) or d-aspartyl (d-Asp) residues during protein aging. These atypical aspartyl residues can interfere with the biological function of the protein and lead to cellular dysfunction. Protein l-isoaspartyl (d-aspartyl) methyltransferase (PIMT) is a repair enzyme that facilitates conversion of l-isoAsp and d-Asp to l-Asp. PIMT deficient mice exhibit accumulation of l-isoAsp in several tissues and die, on average, 12 days after birth from progressive epileptic seizures with grand mal and myoclonus features. However, little is known about the molecular mechanisms by which accumulation of the aberrant residues leads to cellular abnormalities. In this study, we established PIMT-knockdown cells using a short interfering RNA expression system and characterized the resultant molecular abnormalities in intracellular signaling pathways. PIMT-knockdown cells showed significant accumulation of proteins with isomerized residues, compared to control cells. In the PIMT knockdown cells, Raf-1, MEK, and ERK, members of the MAPK cascade, were hyperphosphorylated after EGF stimulation compared to control cells. These results suggest that PIMT repair of abnormal proteins is necessary to maintain normal MAPK signaling. PMID- 18381201 TI - Neuron-specific recombination by Cre recombinase inserted into the murine tau locus. AB - To determine the neuronal function of genes in vivo, the neuron-specific deletion of a target gene in animals is required. Tau, a microtubule-associated protein, is expressed abundantly in neurons but scarcely in glias and other tissues. Therefore, to generate mice that express Cre recombinase in neurons, we inserted Cre recombinase into the tau locus. By crossing these tau-Cre mice with ROSA26 lacZ reporter mice, we observed Cre recombinase activity in the neurons from most of the central nervous system, but not in glias nor in non-neuronal tissues. This neuronal-specific activity appeared during embryogenesis. We further crossed tau Cre mice with rab8 'floxed' mice, and showed that the recombination was nearly complete in the brain, but incomplete or non-detectable in other tissues. Thus, tau-Cre knockin mouse is a useful tool for studying the neuronal function of a gene in vivo. PMID- 18381202 TI - Loss of methylation imprint of Snrpn in postovulatory aging mouse oocyte. AB - Prolonged residence of postovulatory oocyte in the oviduct or prolonged culture in vitro can lead to oocyte aging, which significantly affects pre- and post implantation embryo development. In this study, we employed bisulfite sequencing and COBRA methods to investigate the DNA methylation status of differentially methylated regions (DMRs) of Snrpn and Peg1/Mest, two maternally imprinted genes, in postovulatory oocytes aged in vivo and in vitro. The results showed that Snrpn DMR was clearly demethylated in oocytes aged in vivo at 29h post-hCG and in denuded oocytes aged in vitro for the same time period. However, Peg1/Mest did not show any demethylation in all aged groups at 29h post-hCG. These data indicate that oocytes undergo time-dependent demethylation of Snrpn DMR during the process of postovulatory aging. PMID- 18381203 TI - Transcriptional regulation of human RANK ligand gene expression by E2F1. AB - Receptor activator of nuclear factor kappa B ligand (RANKL) is a critical osteoclastogenic factor involved in the regulation of bone resorption, immune function, the development of mammary gland and cardiovascular system. To understand the transcriptional regulation of RANKL, we amplified and characterized a 1890bp 5'-flanking sequence of human RANKL gene (-1782bp to +108bp relative to the transcription start site). Using a series of deletion mutations of the 1890bp RANKL promoter, we identified a 72bp region (-172 to 100bp) mediating RANKL basal transcriptional activity. Sequence analysis revealed a putative E2F binding site within this 72bp region in the human RANKL promoter. Overexpression of E2F1 increased RANKL promoter activity, while down-regulation of E2F1 expression by small interfering RNA decreased RANKL promoter activity. RT PCR and enzyme linked immunosorbent assays (ELISA) further demonstrated that E2F1 induced the expression of RANKL. Electrophoretic gel mobility shift assays (EMSA) and antibody competition assays confirmed that E2F1 proteins bind to the consensus E2F binding site in the RANKL promoter. Mutation of the E2F consensus binding site in the RANKL promoter profoundly reduced the basal promoter activity and abolished the transcriptional modulation of RANKL by E2F1. These results suggest that E2F1 plays an important role in regulating RANKL transcription through binding to the E2F consensus binding site. PMID- 18381204 TI - KAP1 regulates type I interferon/STAT1-mediated IRF-1 gene expression. AB - Signal transducers and activators of transcription (STATs) mediate cell proliferation, differentiation, and survival in immune responses, hematopoiesis, neurogenesis, and other biological processes. Recently, we showed that KAP1 is a novel STAT-binding partner that regulates STAT3-mediated transactivation. KAP1 is a universal co-repressor protein for the KRAB zinc finger protein superfamily of transcriptional repressors. In this study, we found KAP1-dependent repression of interferon (IFN)/STAT1-mediated signaling. We also demonstrated that endogenous KAP1 associates with endogenous STAT1 in vivo. Importantly, a small-interfering RNA-mediated reduction in KAP1 expression enhanced IFN-induced STAT1-dependent IRF-1 gene expression. These results indicate that KAP1 may act as an endogenous regulator of the IFN/STAT1 signaling pathway. PMID- 18381205 TI - Improvement of sample throughput using fast gas chromatography mass-spectrometry for biochemical diagnosis of organic acid disorders. AB - BACKGROUND: Gas chromatograph mass-spectrometric (GC/MS) method of analysis for urinary organic acids is used for the diagnosis of a variety of metabolic disorders. The method is time-consuming and does not allow for improvements in sample throughput. Although the sample preparation and the data processing have been improved, the long GC/MS analysis time still remains to be problematic. METHODS: The fast-GC/MS method, which utilizes a short microbore capillary GC column and fast temperature programming, was applied to the analysis for urinary organic acids. Urine samples obtained from 15 patients with 9 different disorders and 16 healthy controls were analyzed using conventional GC/MS and fast-GC/MS. RESULTS: Analysis cycle time was shortened from 1 h to 15 min. The automated data system uses retention indices determined by conventional-GC/MS for the identification of 134 organic acids. These retention indices can also be used in data obtained by fast-GC/MS. New fast-GC/MS method with the automated data system gave the same diagnostic results as conventional-GC/MS except for 1 healthy control. CONCLUSIONS: The combined system of fast-GC/MS and the automated data system will be powerful tools in clinical laboratories due to increased sample throughput and reduced analysis costs. PMID- 18381206 TI - Role of endogenous cortisol on Helicobacter pylori colonization. AB - BACKGROUND AND AIMS: Helicobacter pylori infects the gastric mucosa and can lead to chronic gastritis, gastric ulcer and gastric cancer. Colonization of H. pylori in the gastric mucosa is influenced by a variety of host, bacterial and environmental factors. Host defense mechanisms have been affected by endogenous glucocorticoids. We aimed to investigate the relationship between H. pylori and endogenous glucocorticoid. MATERIALS AND METHODS: Forty cases with endoscopically and histologically proven H. pylori and 26 patients who did not have H. pylori on gastric biopsy samples were enrolled in our study. Cortisol was tested from 24-h collected urine samples. RESULTS: H. pylori (+) and H. pylori (-) groups consisted of 40 (28 women, 12 men; aged 44.85+/-12.52 years) and 26 (22 women, 4 men; aged 52.27+/-15.15 years) patients, respectively. Age and gender were similar in both groups. Body mass index, C-reactive protein and erythrocyte sedimentation rate were not statistically different between the two groups (p>0.05). 24-h urine cortisol amount was lower in patients with H. pylori (+) than H. pylori (-) cases. CONCLUSIONS: Present study demonstrates that patients with gastric H. pylori colonization have significantly lower cortisol levels when compared with H. pylori negative cases. There is a negative correlation between H. pylori colonization and urine cortisol output. PMID- 18381207 TI - Reproductive hormones regulate the selective permeability of the blood-brain barrier. AB - Reproductive hormones have been demonstrated to modulate both gap and tight junction protein expression in the ovary and other reproductive tissues, however the effects of changes in reproductive hormones on the selective permeability of the blood-brain barrier (BBB) remain unclear. Age-related declines in BBB integrity correlate with the loss of serum sex steroids and increase in gonadotropins with menopause/andropause. To examine the effect of reproductive senescence on BBB permeability and gap and tight junction protein expression/localization, female mice at 3 months of age were either sham operated (normal serum E2 and gonadotropins), ovariectomized (low serum E2 and high serum gonadotropins) or ovariectomized and treated with the GnRH agonist leuprolide acetate (low serum E2 and gonadotropins). Ovariectomy induced a 2.2-fold increase in Evan's blue dye extravasation into the brain. The expression and localization of the cytoplasmic membrane-associated tight junction protein zona occludens 1 (ZO-1) in microvessels was not altered among groups indicating that the increased paracellular permeability was not due to changes in this tight junction protein. However, ovariectomy induced a redistribution of the gap junction protein connexin-43 (Cx43) such that immunoreactivity relocalized from along the extracellular microvascular endothelium to become associated with endothelial cells. An increase in Cx43 expression in the mouse brain following ovariectomy was suppressed in ovariectomized animals treated with leuprolide acetate, indicating that serum gonadotropins rather than sex steroids were modulating Cx43 expression. These results suggest that elevated serum gonadotropins following reproductive senescence may be one possible cause of the loss of selective permeability of the BBB at this time. Furthermore, these findings implicate Cx43 in mediating changes in BBB permeability, and serum gonadotropins in the cerebropathophysiology of age-related neurodegenerative diseases such as stroke and Alzheimer's disease. PMID- 18381208 TI - Antidepressant-like effects of the delta-opioid receptor agonist SNC80 ([(+)-4 [(alphaR)-alpha-[(2S,5R)-2,5-dimethyl-4-(2-propenyl)-1-piperazinyl]-(3 methoxyphenyl)methyl]-N,N-diethylbenzamide) in an olfactory bulbectomized rat model. AB - The responses of olfactory bulbectomized (OBX) rats to antidepressant treatment are similar to those of depressed patients since chronic administration of an antidepressant reverses OBX-induced behavioral and physiological changes. Previously, using several animal models, it was demonstrated that single treatment with delta-opioid receptor agonists produced an antidepressant-like effect. This study examined the antidepressant effects resulting from subchronic exposure for 8 days to the delta-opioid receptor agonist SNC80 in an OBX rat model of depression. The olfactory bulbs were removed by suction. The emotionality of rats was measured by scoring their responses to given stimuli, i.e., attack, startle, struggle, and fight responses. The OBX rats chronically treated with vehicle for 7 days at 14 days following surgery showed a significant increase in emotionality score and a decrease in the time spent and entries in the open arm of a plus-maze. In the case of OBX rats, these changes were dose- and time-dependently reversed by chronic SNC80 treatment (1-10 mg/kg, s.c.) for 7 days, as same as desipramine (10 mg/kg, i.p.). Moreover, the concentration of 5 HT and its metabolite 5-HIAA in the frontal cortex, hippocampus, and amygdala were decreased in OBX rats, and these changes were also normalized by SNC80 treatment, rather than desipramine treatment. In addition, SNC80 also significantly reversed the loss of TH-positive cells produced by OBX in the dorsal raphe. In conclusion, we demonstrated that subchronic SNC80 treatment could completely reverse OBX-induced behavioral abnormalities and defects in serotonergic function. PMID- 18381209 TI - Maintenance of systemic immune functions prevents accelerated presbycusis. AB - There is no effective therapy for progressive hearing loss such as presbycusis, the causes of which remain poorly understood because of the difficulty of separating genetic and environmental contributions. In the present study, we show that the age-related dysfunctions of the systemic immune system in an animal model of accelerated presbycusis (SAMP1, senescence-accelerated mouse P1) can be corrected by allogeneic bone marrow transplantation (BMT). We also demonstrate that this presbycusis can be prevented; BMT protects the recipients from age related hearing impairment and the degeneration of spiral ganglion cells (SGCs) as well as the dysfunctions of T lymphocytes, which have a close relation to immune senescence. No donor cells are infiltrated to the spiral ganglia, confirming that this experimental system using BMT is connected to the systemic immune system and does not contribute to transdifferentiation or fusion by donor hematopoietic stem cells (HSCs), or to the direct maintenance of ganglion cells by locally infiltrated donor immunocompetent cells. Therefore, another procedure which attempts to prevent the age-related dysfunctions of the recipient immune system is the inoculation of syngeneic splenocytes from young donors. These mice show no development of hearing loss, compared with the recipient mice with inoculation of saline or splenocytes from old donors. Our studies on the relationship between age-related systemic immune dysfunctions and neurodegeneration mechanisms open up new avenues of treatment for presbycusis, for which there is no effective therapy. PMID- 18381210 TI - Effect of the cryoprotectant concentration on the in vitro embryo development and cell proliferation of OPS-vitrified porcine blastocysts. AB - Our objective was to study the effect of the concentration of ethylene glycol (EG) and dimethyl sulfoxide (Me2SO) during vitrification on the development of porcine blastocysts. Vitrification was performed with 0.4 M sucrose and either a Me2SO and EG mixture (15%, 16% and 17% v/v of each) or EG alone (40% v/v), using superfine open pulled straws. Fresh and vitrified blastocysts were cultured for 48 h and the survival and hatching rates were evaluated. Some vitrified and fresh embryos were processed for Hoechst 33342 staining and proliferation cell nuclear antigen (PCNA) inmunolocalization to determine the proliferation index. The survival rate was similar for fresh and vitrified blastocysts, except for blastocysts vitrified using 15% of cryoprotectants, which displayed lower (P < 0.05) survival than fresh blastocysts. Vitrified and fresh blastocysts had a similar cell proliferation index (range: 75.8+/-3.2 to 83.7+/-3). When only hatched blastocysts among groups were compared, the proliferation rate decreased (P < 0.05) after vitrification with 17% of EG-Me2SO. In conclusion, the concentration of EG-Me2SO could be decreased to 16% in the vitrification medium with no reduction of the in vitro developmental ability of the blastocysts. In addition, a 40% EG-based medium can be used for vitrification with similar results to those achieved with a medium containing 16% EG-Me2SO. PMID- 18381211 TI - A role for GPRx, a novel GPR3/6/12-related G-protein coupled receptor, in the maintenance of meiotic arrest in Xenopus laevis oocytes. AB - Progesterone-induced Xenopus laevis oocyte maturation is mediated via a plasma membrane-bound receptor and does not require gene transcription. Evidence from several species suggests that the relevant progesterone receptor is a G-protein coupled receptor (GPCR) and that a second receptor-GPR3 and/or GPR12 in mammals tonically opposes the progesterone receptor. We have cloned a novel X. laevis GPCR, GPRx, which may play a similar role to GPR3/GPR12 in amphibians and fishes. GPRx is related to but distinct from GPR3, GPR6, and GPR12; GPRx orthologs are present in Xenopus tropicalis and Danio rerio, but apparently not in birds or mammals. X. laevis GPRx is mainly expressed in brain, ovary, and testis. The GPRx mRNA increases during oogenesis, persists during oocyte maturation and early embryogenesis, and then falls after the midblastula transition. Microinjection of GPRx mRNA increases the concentration of cAMP in oocytes and causes the oocytes to fail to respond to progesterone, and this block is reversed by co-injecting GPRx with morpholino oligonucleotides. Morpholino injections did not cause spontaneous maturation of oocytes, but did accelerate progesterone-induced maturation. Thus, GPRx contributes to the maintenance of G2-arrest in immature X. laevis oocytes. PMID- 18381212 TI - Terminal end bud maintenance in mammary gland is dependent upon FGFR2b signaling. AB - We previously demonstrated that Fibroblast Growth Factor 10 (FGF10) and its receptor FGFR2b play a key role in controlling the very early stages of mammary gland development during embryogenesis [Mailleux, A.A., Spencer-Dene, B., Dillon, C., Ndiaye, D., Savona-Baron, C., Itoh, N., Kato, S., Dickson, C., Thiery, J.P., and Bellusci, S. (2002). Role of FGF10/FGFR2b signaling during mammary gland development in the mouse embryo. Development 129, 53-60. Veltmaat, J. M., Relaix, F., Le, L.T., Kratochwil, K., Sala, F.G., van Veelen, W., Rice, R., Spencer-Dene, B., Mailleux, A.A., Rice, D.P., Thiery, J.P., and Bellusci, S. (2006). Gli3 mediated somitic Fgf10 expression gradients are required for the induction and patterning of mammary epithelium along the embryonic axes. Development 133, 2325 35.]. However, the role of FGFR2b signaling in postnatal mammary gland development is still elusive. We show that FGF10 is expressed at high level throughout the adipose tissue in the mammary gland of young virgin female mice whereas its main receptor FGFR2 is found mostly in the epithelium. Using a rtTA transactivator/tetracycline promoter approach allowing inducible and reversible attenuation of the FGFR2b signaling throughout the adult mouse, we are now reporting that FGFR2b signaling is also critical during postnatal mammary gland development. Ubiquitous attenuation of FGFR2b signaling in the postnatal mouse for 6 weeks starting immediately after birth is not lethal and leads to minor defects in the animal. Upon dissection of the mammary glands, a 40% reduction in size compared to the WT control is observed. Further examination shows a rudimentary mammary epithelial tree with completely absent terminal end buds (TEBs), compared to a well-branched structure observed in wild type. Transplantation of mammary gland explants into cleared fat pad of wild type mouse recipients indicates that the observed abnormal branching results from defective FGFR2b signaling in the epithelium. We also demonstrate that this rudimentary tree reforms TEBs and resumes branching upon removal of doxycycline suggesting that the regenerative capacities of the mammary epithelial progenitor cells were still functional despite long-term inactivation of the FGFR2b pathway. At the cellular level, upon FGFR2b attenuation, we show an increase in apoptosis associated with a decrease in the proliferation of the mammary luminal epithelium. We conclude that during puberty, there is a differential requirement for FGFR2b signaling in ductal vs. TEBs epithelium. FGFR2b signaling is crucial for the survival and proliferation of the mammary luminal epithelial cells, but does not affect the regenerative potential of the mammary epithelial progenitor cells. PMID- 18381213 TI - Manually-stimulated recovery of motor function after facial nerve injury requires intact sensory input. AB - We have recently shown in rat that daily manual stimulation (MS) of vibrissal muscles promotes recovery of whisking and reduces polyinnervation of muscle fibers following repair of the facial nerve (facial-facial anastomosis, FFA). Here, we examined whether these positive effects were: (1) correlated with alterations of the afferent connections of regenerated facial motoneurons, and (2) whether they were achieved by enhanced sensory input through the intact trigeminal nerve. First, we quantified the extent of total synaptic input to motoneurons in the facial nucleus using synaptophysin immunocytochemistry following FFA with and without subsequent MS. We found that, without MS, this input was reduced compared to intact animals. The number of synaptophysin positive terminals returned to normal values following MS. Thus, MS appears to counteract the deafferentation of regenerated facial motoneurons. Second, we performed FFA and, in addition, eliminated the trigeminal sensory input to facial motoneurons by extirpation of the ipsilateral infraorbital nerve (IONex). In this paradigm, without MS, vibrissal motor performance and pattern of end-plate reinnervation were as aberrant as after FFA without MS. MS did not influence the reinnervation pattern after IONex and functional recovery was even worse than after IONex without MS. Thus, when the sensory system is intact, MS restores normal vibrissal function and reduces the degree of polyinnervation. When afferent inputs are abolished, these effects are eliminated or even reversed. We conclude that rehabilitation strategies must be carefully designed to take into account the extent of motor and/or sensory damage. PMID- 18381214 TI - Costs and benefits of representational change: effects of context on age and sex differences in symbolic magnitude estimation. AB - Studies have reported high correlations in accuracy across estimation contexts, robust transfer of estimation training to novel numerical contexts, and adults drawing mistaken analogies between numerical and fractional values. We hypothesized that these disparate findings may reflect the benefits and costs of learning linear representations of numerical magnitude. Specifically, children learn that their default logarithmic representations are inappropriate for many numerical tasks, leading them to adopt more appropriate linear representations despite linear representations being inappropriate for estimating fractional magnitude. In Experiment 1, this hypothesis accurately predicted a developmental shift from logarithmic to linear estimates of numerical magnitude and a negative correlation between accuracy of numerical and fractional magnitude estimates (r= .80). In Experiment 2, training that improved numerical estimates also led to poorer fractional magnitude estimates. Finally, both before and after training that eliminated age differences in estimation accuracy, complementary sex differences were observed across the two estimation contexts. PMID- 18381215 TI - Hypoxia does not promote naphthalene bioaccumulation in the brown shrimp, Penaeus aztecus. AB - Since increased ventilation is known to be a common strategy used by aquatic animals to cope with hypoxia, we tested in present study the hypothesis that hypoxia can promote the bioaccumulation of naphthalene, a representative polycyclic aromatic hydrocarbon (PAH), in Penaeus aztecus, a penaeid shrimp subject to hypoxia and PAH contamination in the northern Gulf of Mexico. For each of the two naphthalene concentrations, five groups of shrimps were, respectively, subjected to five different conditions, namely, clean seawater under normoxia, seawater containing acetone under normoxia and hypoxia, and seawater containing 10 or 250 microg/L naphthalene under normoxia and hypoxia. Our results suggest that hypoxia does not significantly alter naphthalene bioaccumulation in either the gills or the hepatopancreas of P. aztecus. The absence of a promoting effect of hypoxia on naphthalene bioaccumulation is attributed to the increased disposition of naphthalene when the shrimps are subjected to hypoxia. PMID- 18381216 TI - The toxic benthic dinoflagellate Ostreopsis ovata: quantification of proliferation along the coastline of Genoa, Italy. AB - Toxic harmful algal bloom (HAB) occurrence is becoming more frequent and problematic in highly urbanized coastal zones. In summer 2005 along the urbanized Genoa coastline (Ligurian Sea, North western Mediterranean Sea), local first aid stations treated about 200 people, who all showed similar symptoms following exposure to marine aerosols. The link with proliferation of Ostreopsis ovata was made, and it highlighted for the first time, the risks that benthic HABs may represent in highly urbanised temperate areas. Subsequently, a specific monitoring plan was designed and implemented in the same area in July 2006, before the first signs of Ostreopsis proliferation were detected. Here we report on this quantification of an Ostreopsis ovata bloom in the Ligurian Sea. Cells were quantified both in the water column and in the epiphytic community on macrophytes. Our results suggest a role of sea water temperature and weather conditions in favouring bloom development. PMID- 18381217 TI - Relationships between hydrosedimentary processes and occurrence of mercury resistant bacteria (merA) in estuary mudflats (Seine, France). AB - The Seine estuary (France) is one of the world's macrotidal systems that is most contaminated with heavy metals. To study the mercury-resistant bacterial community in such an environment, we have developed a molecular tool, based on competitive PCR, enabling the quantification of Gram-negative merA gene abundance. The occurrence of the Gram-negative merA gene in relation with the topology (erosion/deposit periods) and the mercury contamination of three contrasted mudflats was investigated through a multidisciplinary approach and compared with a non-anthropized site (Authie, France). The higher abundance of the Gram-negative merA gene in the Seine estuary mudflats indicates a relationship between the degree of anthropization and the abundance of the merA gene in the mudflat sediments. In the Seine mudflats, the maxima of abundance are always located in fresh sediment deposits. Therefore, the abundance is closely related with the hydrosedimentary processes, which thus seem to be determining factors in the occurrence of the Gram-negative merA gene in the surface sediments of the Seine's mudflat. PMID- 18381218 TI - Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis). AB - Aliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed. Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2-3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors. PMID- 18381219 TI - Toxicogenomic analysis of immune system-related genes in Japanese flounder (Paralichthys olivaceus) exposed to heavy oil. AB - Heavy oil contamination is one of the most important environmental issues. Toxicities of polycyclic aromatic hydrocarbons (PAHs), including immune toxicities, are well characterized, however, the immune toxic effects of heavy oil, as a complex mixture of PAHs, have not been investigated. In the present study, we selected Japanese flounder (Paralichthys olivaceus) as a model organism, and observed alteration of immune function by the exposure to heavy oil. To analyze the expression profiles of immune system-related genes, we selected 309 cDNAs from our flounder EST library, and spotted them on a glass slide. Using this cDNA array, alteration of gene expression profiles was analyzed in the kidneys of flounders exposed to heavy oil. Six Japanese flounders (mean body weight: 197 g) were acclimated to laboratory conditions at 19-20 degrees C. Three fish were exposed to heavy oil C (bunker C) at a concentration of 3.8 g/L for 3 days, and the others were kept in seawater without heavy oil and used as the control. After the exposure period, the fish were transferred into control seawater and maintained for 4 days, and then they were dissected and their kidneys were removed. Total RNA was extracted from the kidney samples to use in gene expression analyses. The microarray detected alteration of immune system related genes in the kidneys of heavy oil-exposed flounders, including down regulation of immunoglobulin light chain, CD45, major histocompatibility complex class II antigens and macrophage colony-stimulating factor precursor, and up regulation of interleukin-8 and lysozyme. These results suggest that pathogen resistance may be weakened in heavy oil-exposed fish, causing a subsequent bacterial infection, and then proinflammatory genes may be induced as a defensive response against the infection. Additionally, we found candidate genes for use as biomarkers of heavy oil exposure, such as N-myc downstream regulated gene 1 and heat shock cognate 71 kDa proteins. PMID- 18381220 TI - Amelioration of marine farming impact on the benthic environment using artificial reefs as biofilters. AB - An in situ monitoring of the sediment characteristics and macrobenthic communities was undertaken at a marine fish culture site in subtropical waters of Hong Kong before and after the deployment of biofilters which were made of cement concrete artificial reef (AR) structures. According to the distance to the boundary of the fish cages, 6 points were selected as sampling stations: 2 at the fish cages, 2 near the boundary of the fish culture area, and 2 reference sites further away from the culture area. Bimonthly sediment samples were collected for analysis of silt-clay fraction (SCF), moisture content (MC), total organic carbon (TOC), total Kjeldahl nitrogen (TKN) and total phosphorus (TP). The macrobenthos (>0.5mm) present in the sediment were sorted, identified and enumerated. TOC, TKN and TP levels at the fish cage stations were consistently higher than those at the reference stations over the 1-year pre-AR and 2-year post-AR deployment monitoring period. The diversity of macrofauna was significantly reduced at the fish cage stations relative to the reference sites. The intermediary stations near the fish culture area showed a transitional state of disturbance. Over the 2 year post-AR deployment period, TOC, TKN and TP showed a decreasing trend at the fish cage and intermediary stations. More diverse macrofaunal communities were recorded at the fish cage stations, with species diversity H'increasing from 0-1 at the beginning of the AR deployment to H'>2 at the end of the study. The present results demonstrated that artificial reefs can improve the benthic abiotic environment and biotic conditions beneath fish rafts which are deteriorated due to farming activities. PMID- 18381221 TI - Immuno-localisations (GSSP) of subcellular accumulation sites of phenanthrene, aroclor 1254 and lead (Pb) in relation to cytopathologies in the gills and digestive gland of the mussel Mytilus edulis. AB - Cell and tissue pathology of both, gill and digestive tissue, has been the subject of many studies for the elucidation of contaminant-induced biological effects. In the present study, cellular pathological alterations were linked to subcellular sites of chemical accumulation in gills and digestive gland tissues. For this purpose, mussels (Mytilus edulis) were exposed to the organic contaminants aroclor 1254 (PCB) (20 microg/L), phenanthrene (PAH) (150 microg/L) or the metal lead (Pb) (2.5mg/L). The localization of chemicals at the subcellular level was analysed by an antibody-based detection system (GSSP) by the use of commercially available antibodies specifically directed against the chemicals. Pathological changes were analysed in parallel in identical samples by transmission electron microscopy. After exposure to the different contaminants, cell organelles such as mitochondria, the endo-lysosomal system as well as endoplasmic reticulum showed clear evidence of chemically-induced alterations. Large numbers of crystalloid inclusions were found in mitochondria and in autophagic lysosomes as well as multi-lamellated whorls after PAH and aroclor exposure. Immunocytochemical detection of the chemicals showed their accumulation inside of various cell organelles such as lysosomes, mitochondria, and nuclei. Additionally, chemicals were localized in association to membranes, cilia and microvilli of gill and digestive gland cells. Furthermore, the chitinous rod and mucus secretions of gill epithelial cells were positively labelled for contaminants indicating their role in protection. Localization of contaminants by immuno-detection in combination with pathological diagnosis gives insights into the cellular targets of chemical attack. PMID- 18381222 TI - Cellular energy allocation in the Arctic sea ice amphipod Gammarus wilkitzkii exposed to the water soluble fractions of oil. AB - Increasing offshore oil and gas activities in the European Arctic has raised concerns of the potential anthropogenic impact of oil-related compounds on the polar marine ecosystem. We measured cellular energy allocation (CEA) in the sea ice amphipod Gammarus wilkitzkii after exposure for one month to the water soluble fraction (WSF) of oil. The CEA biomarker measures the energy budget of organisms by biochemically assessing changes in carbohydrates, protein and lipid content as well as the electron transport system activity. A significantly higher protein content was observed in the medium dose compared to controls, while the total energy budget was not affected in G. wilkitzkii. This indicates that parts of the energy budget of G. wilkitzkii, which is a key species in the Arctic ecosystem, is affected by a WSF of oil. PMID- 18381223 TI - Cloning and mRNA expression analysis of an ABCG2 (BCRP) efflux transporter in rainbow trout (Oncorhynchus mykiss) liver and primary hepatocytes. AB - As has been recently demonstrated in mammals, apart from the P-glycoprotein (Pgp, ABCB) and the MRP-like proteins (MRPs, ABCC), another efflux transporter - the BCRP (ABCG2) - expressed in polarized epithelial cells of different tissues, is involved in regulation of intestinal absorption and biliary excretion of potentially toxic xenobiotics. However, no study has been directed towards identification of BCRP (ABCG2) in fish species. In the present study we have cloned the full ABCG2 transcript from rainbow trout (Oncorhynchus mykiss) liver, showing a high sequence identity (60%) to human ABCG2 gene. Using Real-Time PCR we measured relative expression of ABCG2 in trout liver and primary hepatocytes and compared these expression levels to the expression of other ABC transporters expressed in apical membrane of hepatocytes (ABCB1, ABCB11 and ABCC2). ABCG2 expression was 6-fold higher than ABCC2 and almost 42 fold higher than ABCB1, indicating that the ABCG2 probably plays a significant role in the overall disposition and excretion of xenobiotics in fish. PMID- 18381224 TI - Is delta-aminolevulinic acid dehydratase activity in bivalves from south-west Iberian Peninsula a good biomarker of lead exposure? AB - The coast of Huelva is considered as a heavily contaminated area where the Tinto and Odiel rivers discharge after running through a metalliferous mining area in the Iberian Pyrite Belt and end in common estuary called Ria of Huelva. Lead is a highly toxic and widely distributed element in the aquatic environment; therefore there is a great interest in assessing the impact of this contaminant on aquatic organisms. To study the bioavailability and sources of lead, the bivalve species Chamelea gallina was collected from six sampling sites along the coast of Huelva. Besides the effect of this metal on the enzyme activity, delta-aminolevulinic acid dehydratase (ALA-D) was determined in the whole tissue. Results show the inverse relationship between the lead concentration and the activity of ALA-D measured in whole tissues (r2=0.7). Individuals collected from the Ria of Huelva estuary had the highest levels of total lead concentrations and the lowest activity of ALA-D. Lead isotope analyses demonstrated the different sources of lead contamination and also confirmed the influence of Ria of Huelva on the metal input to the marine environment. PMID- 18381225 TI - Preliminary analysis of the forces on the thoracic cage of patients with pectus excavatum after the Nuss procedure. AB - BACKGROUND: The Nuss procedure corrects pectus excavatum using a pre-bent bar that generates stress on the chest wall. To investigate the biomechanical effects after the Nuss procedure, we designed a three-dimensional finite element analysis model to analyze the distribution of stress and strain induced in the chest wall. METHODS: Three patients with pectus excavatum aged 8, 7, and 7 years, were enrolled in this study. The greatest upward displacements of their sternums after the operation were measured from computed tomography images and chest X-ray films. Based on these displacements, we constructed three finite element analysis models for analyzing biomechanical changes in the thoracic cage after the Nuss procedure. FINDINGS: The simulation results indicated that greatest strain occurred at the third through seventh cartilages, especially where they join the sternum and ribs. A high bilateral stress distribution was also found over the backs of the third to the seventh ribs near the vertebral column. INTERPRETATION: The stress and strain induced by the Nuss procedure can be analyzed using our finite element analysis model. Although the stress and strain may have some influence on chest and spine development, a more detailed finite element analysis model is recommended for future study to improve the accuracy of our simulation results. PMID- 18381226 TI - Perceptual-attentional and motor-intentional bias in near and far space. AB - Spatial bias demonstrated in tasks such as line-bisection may stem from perceptual-attentional (PA) "where" and motor-intentional (MI) "aiming" influences. We tested normal participants' line bisection performance in the presence of an asymmetric visual distracter with a video apparatus designed to dissociate PA from MI bias. An experimenter stood as a distractor to the left or right of a video monitor positioned in either near or far space, where participants viewed lines and a laser point they directed under (1) natural and (2) mirror-reversed conditions. Each trial started with the pointer positioned at either the top left or top right corner of the screen, and alternated thereafter. Data analysis indicated that participants made primarily PA leftward errors in near space, but not in far space. Furthermore, PA, but not MI, bias increased bilaterally in the direction of distraction. In contrast, MI, but not PA, bias was shifted bilaterally in the direction of startside. Results support the conclusion that a primarily PA left sided bias in near space is consistent with right hemisphere spatial attentional dominance. A bottom-up visual distractor specifically affected PA "where" spatial bias while top-down motor cuing influenced MI "aiming" bias. PMID- 18381227 TI - Comparison of grain from corn rootworm resistant transgenic DAS-59122-7 maize with non-transgenic maize grain in a 90-day feeding study in Sprague-Dawley rats. AB - DAS-59122-7 (59122) is a transgenic maize (Zea mays L.) that contains genes encoding Cry34Ab1 and Cry35Ab1 proteins from Bacillus thuringiensis Berliner strain 149B1 and phosphinothricin acetyltransferase (PAT) protein from Streptomyces viridochromogenes. Expression of these proteins in planta confers resistance to corn rootworms and other Coleopteran parasites and tolerance to herbicides containing glufosinate ammonium, respectively. In the current study, processed flours from 59122 maize grain or its near isogenic control line (091) were used at two concentrations (50% and 70% wt/wt) to produce diets that were fed to rats for 90 days in accordance with Chinese toxicology guidelines (GB15193.13-2003). A commercial AIN93G diet was used as an additional negative control. No significant differences in body weight and feed utilization were observed between rats consuming diets formulated with 59122 and 091 Control corn. Statistical differences (p<0.05) were observed in certain hematology and serum chemistry response variables between rats consuming diets formulated with 59122 or 091 Control flour compared to AIN93G diet. However, the mean value of these response variables in the 59122 groups were not statistically different from those observed in diets formulated with corresponding high and low concentrations of the flour from the 091 Control maize grain. Therefore, the statistical differences were considered to be related to consumption of diets containing high concentrations of maize flour (compared to AIN93G diets) regardless of source rather than to consumption of flour from 59122 maize grain. The results from this study demonstrated that 59122 maize grain is as safe as non-transgenic maize grain. PMID- 18381228 TI - Genotoxicity studies on green tea catechin. AB - The beneficial effects of tea catechins are well documented. We evaluated the genotoxic potential of a green tea catechin preparation using established genotoxicity assays, including a bacterial reverse mutation assay (Ames test), a chromosomal aberration assay in cultured Chinese hamster lung cells (CHL/IU), a mouse lymphoma L5178Y/tk assay, and a bone marrow micronucleus (MN) assay in ICR CD mice and SD rats. No significant increases in the number of revertant colonies were observed in the Ames test, but positive responses were observed in two in vitro assays: the chromosomal aberration assay and mouse lymphoma L5178/tk assay. However, the in vivo study demonstrated no significant increase in micronucleated polychromatic erythrocytes (MNPCE) in the bone marrow of both ICR CD mice and SD rats administered a high dose of the green tea catechin preparation up to 2000mg/kg. Combined with favorable epidemiological information suggesting a chemopreventive effect of tea catechins on carcinogenesis, we conclude that green tea catechin presents no significant genotoxic concern under the anticipated conditions of use. These results are consistent with other genotoxicity studies of tea catechins, which show minimal, if any, genotoxic potential. PMID- 18381229 TI - Evaluation of subchronic toxicity of dietary administered Cry1Ab protein from Bacillus thuringiensis var. Kurustaki HD-1 in F344 male rats with chemically induced gastrointestinal impairment. AB - Bacillus thuringiensis (Bt) proteins are developed for genetically modified crops and the Bt proteins demonstrate no evidence of toxicity by the oral route in traditional animal models. However, the possible toxicity of Bt proteins under conditions of reduced gastric acid secretion and/or small intestinal damage has not been investigated. In the present study, we therefore evaluated following four F344 rat groups with a purified Bt protein Cry1Ab from B. thuringiensis var. Kurustaki HD-1. Gastrointestinal impairment (GI) alone and GI+Bt protein fed (GI+Bt) groups were given i.p. injections of famotidine to reduce gastric acid secretion twice a day at 30mg/kg body weight in weeks 2 and 4. GI and GI+Bt groups were additionally fed diets containing 80ppm indomethacin for induction of intestinal damage during weeks 1 and 3. Bt alone and GI+Bt groups were also fed diet containing Bt protein Cry1Ab at a concentration of 10ppm in weeks 2 and 4. A no treatment control group was also included. At the end of week 4, all animals were euthanized under ether anesthesia, blood samples were collected for hematology and serum biochemistry and a complete necropsy was performed. No significant changes indicative of toxicity of the Bt protein Cry1Ab used here were noted with any of the parameters investigated. In conclusion, no significant toxicological effects were detected in this subchronic gastrointestinal impairment rat model. PMID- 18381230 TI - Mechanism of apoptosis induced by copper in PC12 cells. AB - Copper, an essential trace element, induces apoptosis in mammalian cells. However, the precise mechanism of copper-induced apoptosis is still unclear. In this study, to determine the apoptotic pathway initiated by copper treatment, apoptotic factors such as Bax, Bad and Bcl-2, and the caspase family in PC12 cells treated with copper were measured by Western blot and RT-PCR analyses. The expression of Bax, Bad, cytochrome c and caspases 3 and 9 were increased by copper treatment. From these results, two pathways for copper-induced apoptosis were suggested. At first, an increase of Bax induces the release of cytochrome c from the mitochondria into the cytoplasm owing to binding to apoptotic activating caspase 9 leading to the activation of caspases 3. In the other pathway an increase of Bax and reactive oxygen species activates the release of AIF from the mitochondria. The AIF induces apoptosis via a caspase-independent pathway. PMID- 18381231 TI - Cytogenetic and molecular aspects of gastric cancer: clinical implications. AB - Gastric cancer is of major importance world-wide being the second most common cause of cancer-related death in the world. According to Lauren's histological classification gastric cancer is divided in two groups, the better differentiated intestinal carcinomas and the poorly differentiated diffuse-type cancers. The genetic changes underlying the initiation and progression of gastric cancer are not well defined. Gastric carcinogenesis is a multistep process involving a number of genetic and epigenetic factors. Although it has been proposed that different genetic pathways exist for differentiated and undifferentiated carcinomas, the two histological subtypes of gastric cancer share some common genetic alterations. Currently, tumor histology and pathologic stage are the major prognostic variables used in the clinical practice for gastric cancer patients. However, it is known that tumors with similar morphology may differ in biological aggressiveness, prognosis and response to treatment. Molecular genetic analysis of gastric cancer revealed a number of associations of certain genetic changes with pathological features, tumor biological behavior and prognosis of gastric cancer patients, suggesting that these genetic abnormalities might play an important role in gastric tumorigenesis. Increasing evidence suggests that the molecular genetic changes could be helpful in the clinical setting, contributing to prognosis and management of patients. Regarding epigenetic events in gastric tumorigenesis, a number of methylating markers have been proposed for risk assessment, prognostic evaluation and as therapeutic targets. However, further research is required in order to systematically investigate the genetic changes in gastric cancer estimating also their usefulness in the clinical practice. A good understanding of the genetic changes underlying gastric carcinogenesis may provide new perspectives for prognosis and screening of high risk individuals. Some of the genetic alterations could definitely improve tumor classification and management of gastric cancer patients. Also, based on molecular data identified in gastric cancer novel therapeutics might help to improve the treatment of this disease. PMID- 18381232 TI - Signaling and regulatory mechanisms of integrin alphavbeta6 on the apoptosis of colon cancer cells. AB - Considerable researches have been done about integrin alphanubeta6 and carcinomas, but little information has been shown about the relationship between integrin alphanubeta6 and apoptosis. In this study, we investigated the apoptosis and its related signal pathways to integrin alphavbeta6 in colon cancer cells. After we blocked the function of integrin alphavbeta6 in HT29 cells used the monoclonal antibody, the apoptotic cells increased markedly. Meanwhile, cytochrome C released from mitochondria into cytosol, Bcl-2 decreased while Bax increased significantly, and Fas and Fas-ligand had no change. The activities of caspase-3 and caspase-9 increased, while caspase-8 remained no change. Moreover, the expression of phosphorylated extracellular signal-related kinase (P-ERK) decreased. We confirmed that integrin alphavbeta6 acted as an important role in inhibiting apoptosis in colon cancer cells, and the signaling involved the mitochondrial pathway. PMID- 18381233 TI - A-kinase anchoring protein mediates TRPV1 thermal hyperalgesia through PKA phosphorylation of TRPV1. AB - Certain phosphorylation events are tightly controlled by scaffolding proteins such as A-kinase anchoring protein (AKAP). On nociceptive terminals, phosphorylation of transient receptor potential channel type 1 (TRPV1) results in the sensitization to many different stimuli, contributing to the development of hyperalgesia. In this study, we investigated the functional involvement of AKAP150 in mediating sensitization of TRPV1, and found that AKAP150 is co expressed in trigeminal ganglia (TG) neurons from rat and associates with TRPV1. Furthermore, siRNA-mediated knock-down of AKAP150 expression led to a significant reduction in PKA phosphorylation of TRPV1 in cultured TG neurons. In CHO cells, the PKA RII binding site on AKAP was necessary for PKA enhancement of TRPV1 mediated Ca2+-accumulation. In addition, AKAP150 knock-down in cultured TG neurons attenuated PKA sensitization of TRPV1 activity and in vivo administration of an AKAP antagonist significantly reduced prostaglandin E2 sensitization to thermal stimuli. These data suggest that AKAP150 functionally regulates PKA mediated phosphorylation/sensitization of the TRPV1 receptor. PMID- 18381234 TI - A meta-analysis and systematic review of thalidomide for patients with previously untreated multiple myeloma. AB - A systematic review and meta-analysis was performed to determine the efficacy and toxicity of thalidomide in previously untreated patients with myeloma. Medline, Embase, Cochrane Controlled Trials Register, and abstracts from the American Society of Hematology and the American Society of Clinical Oncology were searched for randomized controlled trials (RCTs) of either induction or maintenance thalidomide in adults with previously untreated myeloma. Nine RCTs of induction thalidomide, three RCTs of maintenance thalidomide, and one RCT of induction and maintenance thalidomide were identified, involving a total of 4144 subjects. When thalidomide was added to standard, non-transplantation myeloma therapy, overall survival (OS) improved (HR 0.67; 95% CI 0.56-0.81). When thalidomide was given as maintenance following autologous transplantation (ASCT), there was a trend to improved OS (HR 0.61, 95% CI 0.37-1.01); when the only trial which combined induction and maintenance thalidomide was excluded from this analysis, a significant survival advantage emerged (HR 0.49, 95% CI 0.32-0.74). The relative risk of venous thromboembolism (VTE) with induction thalidomide was 2.56 (95% CI 1.88-3.49). A meta-analysis of trials/sub-groups administering low molecular weight heparin (LMWH) as VTE prophylaxis, suggested a persistently increased relative risk of VTE with induction thalidomide (RR 1.54, 95% CI 1.07-2.22). The relative risk of VTE was substantially lower, but still elevated, when thalidomide was given as maintenance therapy following ASCT (RR 1.95, 95% CI 1.15 3.30). In summary, thalidomide appears to improve the overall survival of patients with newly diagnosed myeloma both when it is added to standard, non transplantation therapy, and when it is given as maintenance therapy following ASCT. However, thalidomide is associated with toxicity, particularly a significantly increased risk of VTE. PMID- 18381235 TI - Effect of lead on proliferation and neural differentiation of mouse bone marrow mesenchymal stem cells. AB - Bone marrow-mesenchymal stem cells (MSCs) are considered to be an ideal source of stem cells for assessing the effects of environmental toxins on the proliferation, multipotency and differentiation of adult stem cells. The aim of this study was to investigate the effect of lead on the proliferation and neuronal differentiation of murine MSCs. MTT assay used in this study revealed that while the proliferation of MSCs is sensitive to higher than 10 microM lead, a 50% reduction in the rate of their proliferation can be achieved in the presence of 60 microM lead. The results of immunocytochemistry and RT-PCR showed that beta-mercaptoethanol induced-neuronal differentiation is also reduced after the treatment of MSCs by 60 microM lead. Furthermore, the comet assay analysis of MSCs showed a substantial increase in DNA damage in the lead treated cells compared to the control. In conclusion our results revealed for the first time that lead is not only cytotoxic to the survival and proliferation of MSCs but also inhibits their differentiation to neurons in a dose-dependant manner. Therefore, MSCs appear to be an alternative method for assessing the cytotoxic effects of such environmental hazards. PMID- 18381236 TI - A potential paradox in prostate adenocarcinoma progression: oestrogen as the initiating driver. AB - One in 10 men in the developed world will present with prostate cancer (CaP), and in an ageing population developing strategies for its chemoprevention or treatment is of significance. For decades, androgen ablation has remained the frontline treatment for CaP that is no longer organ-confined and thus deemed surgically inoperable. Orchidectomy or drug-induced reduction of serum testosterone levels with the consequent removal of growth-promoting effects in the prostate is the driving rationale for this regimen. However, resistance often develops within a few months to years and androgen-insensitive tumours develop. In recent years, there has been an increasing focus on chemoprevention with agents such as finasteride being employed to reduce the risk of developing CaP. Significantly, such chemoprevention strategies are also based on 5alpha-reductase inhibition thus reducing intraprostatic dihydrotestosterone levels. Although there may be an overall reduction in CaP incidence in cohorts using such chemoprevention, in a subset of users who do develop this pathology there results a more aggressive, higher-grade disease. There have also been suggestions regarding the protective role of androgens against high-grade CaP. This leads to the intriguing notion that 17beta-oestradiol (E2) may be an initiating driver of CaP; in fact, in old studies in which CaP was induced in rodents, E2 often accelerated the effect of the carcinogen. Might certain chemoprevention strategies or androgen ablation result in a systemic feedback loop in hormone synthesis or metabolism? If so, elevated serum E2 levels could result in its increased conversion to genotoxic catechol oestrogens in target tissues such as the prostate. Paradoxically, if E2 were to be an initiating factor in CaP, anti oestrogens might be an overlooked treatment or chemoprevention strategy. PMID- 18381237 TI - Cancer-related fatigue. AB - Fatigue is a common symptom in patients with cancer and in disease-free survivors. It has a significant impact on the quality of life. Although subjective fatigue is often related to objective changes in physical functioning or impaired performance status, the two phenomena are not synonymous and need to be distinguished. A number of robust and reliable assessment instruments to measure fatigue severity are now available and criteria for cancer-related fatigue syndrome have been proposed. The underlying mechanisms and pathophysiology of cancer-related fatigue are unclear. Management strategies include the use of psycho-educational interventions, exercise programmes and pharmacological treatments. The best evidence for the effectiveness of drug treatments is for the haematopoietic agents in anaemic patients undergoing chemotherapy and for methylphenidate in an on-treatment population. PMID- 18381238 TI - Novel uracil-based 2-aminoanilide and 2-aminoanilide-like derivatives: histone deacetylase inhibition and in-cell activities. AB - A novel series of non-hydroxamate HDAC inhibitors (HDACi) showing a uracil group at the left and a 2-aminoanilide/2-aminoanilide-like portion at the right head have been reported. In particular, the new compounds incorporating a 2 aminoanilide moiety behaved as class I-selective HDACi. Compound 8, the most potent and class I-selective, showed weak apoptosis (higher than MS-275) joined to cytodifferentiating activity on U937 cells. Surprisingly, the highest differentiation was observed with 13, through an effect that seems to be unrelated to HDAC inhibition. PMID- 18381240 TI - Synthesis, cytotoxic activity, and SAR analysis of the derivatives of taxchinin A and brevifoliol. AB - Twenty-one derivatives of taxchinin A (1) and brevifoliol (2) were synthesized and evaluated for cytotoxicity against human non-small lung cancer (A549) cell line. Nine derivatives showed potent activity with IC(50) values from 0.48 to 6.22microM. 5-Oxo-13-TBDMS-taxchinin A (11) and 5-oxo-13,15-epoxy-13-epi taxchinin A (15) are the most potent derivatives, with IC(50) at 0.48 and 0.75microM, respectively. The structure-activity relationship (SAR) of these compounds established that exocyclic unsaturated ketone at ring C is the key structural element for the activity, while the alpha,beta-unsaturated ketone positioned at ring A has no effect for the activity. The significant cytotoxicity of derivatives 11 and 15 may be due to the conformational change in the taxane rings. The 3D-QSAR study was conducted on this series of compounds, which provided optimal predictive comparative molecular field (CoMFA) model with cross validated r(2) (q(2)) value of 0.64. PMID- 18381239 TI - Synthesis and biological evaluation of histone deacetylase inhibitors that are based on FR235222: a cyclic tetrapeptide scaffold. AB - We outline the synthesis of six novel derivatives that are based on a recently discovered HDAC inhibitor FR235222. Our work is the first report utilizing a novel binding element, guanidine, as metal coordinators in HDAC inhibitors. Further, we demonstrate that these compounds show cytotoxicity that parallels their ability to inhibit deacetylase activity, and that the most potent compounds maintain an L-Phe at position 1, and a D-Pro at position 4. Both inhibition of HDAC activity and cytotoxicity against the pancreatic cancer cell line BxPC3 are exhibited by these compounds, establishing that a guanidine unit can be utilized successfully to inhibit HDAC activity. PMID- 18381241 TI - Molecular modeling and dynamics simulations of PNP from Streptococcus agalactiae. AB - This work describes for the first time a structural model of purine nucleoside phosphorylase from Streptococcus agalactiae (SaPNP). PNP catalyzes the cleavage of N-ribosidic bonds of the purine ribonucleosides and 2-deoxyribonucleosides in the presence of inorganic orthophosphate as a second substrate. This enzyme is a potential target for the development of antibacterial drugs. We modeled the complexes of SaPNP with 15 different ligands in order to determine the structural basis for the specificity of these ligands against SaPNP. The application of a novel empirical scoring function to estimate the affinity of a ligand for a protein was able to identify the ligands with high affinity for PNPs. The analysis of molecular dynamics trajectory for SaPNP indicates that the functionally important motifs have a very stable structure. This new structural model together with a novel empirical scoring function opens the possibility to explorer larger library of compounds in order to identify the new inhibitors for PNPs in virtual screening projects. PMID- 18381243 TI - The reliability of electromechanical delay and torque during isometric and concentric isokinetic contractions. AB - The purpose of this investigation was to examine the intra-subject reliability of electromechanical delay (EMD) and torque of the dominant and non-dominant elbow flexors during isometric and isokinetic muscle contractions repeated over five consecutive days. Eleven volunteers that were unfamiliar with isokinetic dynamometry participated in this study and were asked to attend the laboratory on five consecutive days. An isokinetic dynamometer was used to exercise the elbow flexors under isometric, slow (60 degrees s(-1)) and fast (210 degrees s(-1)) isokinetic conditions; surface electromyography was recorded from the belly of biceps brachii and the signal was synchronised with the dynamometer to determine EMD. Intra-subject reliability for all measures was good (CV range, 3.1-6.5%) with no discernable difference between the dominant and non-dominant arms during isometric and isokinetic conditions. In addition, there was little difference in EMD and torque variability between the dominant and non-dominant arms which may have applications for clinicians and future research design when monitoring and investigating human muscle function. These data provide researchers and clinicians with an indication of the magnitude of change that is required to elucidate the presence of a meaningful change to muscle function in the elbow flexors. PMID- 18381242 TI - BDNF activates CaMKIV and PKA in parallel to block MAG-mediated inhibition of neurite outgrowth. AB - The environment of the adult CNS prevents axonal regeneration after injury. This inhibition of axonal regeneration can be blocked by elevating cAMP. Previously, we showed that the cAMP pathway can be activated via pre-treatment with neurotrophins and requires activation of several signaling pathways which converge at activation of the transcription factor, CREB. Here, we show that calcium/calmodulin-dependent kinase IV (CaMKIV) is necessary for the neurotrophin induced phosphorylation of CREB and the block of myelin-mediated inhibition of axonal growth. Pharmacological inhibition of CaMKIV or over-expression of a dominant-negative mutant form of CaMKIV blocks the neurotrophin effect. Interestingly, CaMKIV activation is not necessary if cAMP levels is already elevated. Finally, calcium flux from intracellular stores is necessary for this CaMKIV signaling. These results demonstrate that CaMKIV is another player in the neurotrophin-induced signaling which leads to axonal regeneration and therefore, is a potential target for therapeutic intervention following injury to the adult CNS. PMID- 18381244 TI - Identification of a novel frameshift mutation at codon 53 (-T) in the beta-globin gene causing dominantly inherited beta-thalassemia in a Chinese Miao family. AB - beta-thalassemia, one of the most common inherited disorders of hemoglobin synthesis in the world, is genetically heterogeneous with over 200 different beta globin mutations worldwide. In this study, we describe a novel frameshift beta thalassemia mutation at codon (cd) 53 (-T) in exon 2 of the beta-globin gene in a Chinese Miao family. In this family, all seven heterozygotes with this mutation presented with moderate anemia, jaundice, splenomegaly and elevated hemoglobin A2 levels. None of them had been transfused or carried any other known alpha/beta globin mutation. Pedigree analysis indicated an autosomal dominant inheritance pattern in this family. Two new haplotypes "----+-+" and "--+++-+" were identified by restriction fragment length polymorphism (RFLP) haplotype analysis. The former was associated with the cd53 (-T) mutation and the latter only existed in one family member. Thus, a novel frameshift cd53 (-T) mutation may lead to mild thalassemia intermedia even though there is no statistically significant difference in beta-globin messenger RNA (mRNA) level between six heterozygotes and six normal subjects. PMID- 18381245 TI - Diabetic osteopathy and the IGF system in the Goto-Kakizaki rat. AB - OBJECTIVE: Diabetes mellitus is associated with an increased risk of osteopenia, fracture and Charcot arthropathy. Abnormalities of the IGF system commonly observed in diabetes may underlie this "diabetic osteopathy" as IGF-I and its binding proteins (IGFBPs) have been shown to affect osteoblast and osteoclast activity. DESIGN: In type-2 diabetic and control rats we analyzed IGF-I and IGFBP 1 and -4 levels in serum, and notably, also the IGF-I levels in cortical bone, ankles and vertebrae by immunoassays. Osteopathy was assessed by radiography and dual energy X-ray absorptiometry. RESULTS: In the diabetic rats IGF-I was significantly reduced in serum and diaphyseal bone while IGFBP-1 and IGFBP-4 were increased in serum. The periosteal and endosteal diameters were increased in the diaphysis of humerus and tibia (changes similar to those in elderly humans) while bone mineral density was reduced in long bone metaphyses and vertebrae. CONCLUSIONS: Our study demonstrates both systemic and local disturbances of the IGF-system in rats with type-2 diabetes, consistent with the observed enhanced endosteal erosion in long bone diaphyses, and osteopenia in metaphyses and vertebrae. Whether similar IGF-system changes contribute to osteopathy in patients with diabetes and if treatment of diabetes can reverse the osteopathy has yet to be explored. PMID- 18381246 TI - Objective post-mortem diagnosis of chronic alcohol abuse--a review of studies on new markers. AB - Identification of chronic excessive alcohol consumption in living and deceased individuals is a fundamental task in forensic pathology. Reliable methods for post-mortem diagnosis of chronic alcohol abuse are required because morphological findings are unspecific and ante-mortem data are often unreliable. In clinical practice, several biochemical markers indirectly demonstrating chronic alcohol abuse are employed, but thus far these methods have not been used in routine post mortem investigations. We reviewed publications in which these markers have been applied to autopsy material. Based on this review, some of these biochemical parameters are useful in post-mortem diagnostics, although further systematic research is required. PMID- 18381247 TI - MRI denoising using non-local means. AB - Magnetic Resonance (MR) images are affected by random noise which limits the accuracy of any quantitative measurements from the data. In the present work, a recently proposed filter for random noise removal is analyzed and adapted to reduce this noise in MR magnitude images. This parametric filter, named Non-Local Means (NLM), is highly dependent on the setting of its parameters. The aim of this paper is to find the optimal parameter selection for MR magnitude image denoising. For this purpose, experiments have been conducted to find the optimum parameters for different noise levels. Besides, the filter has been adapted to fit with specific characteristics of the noise in MR image magnitude images (i.e. Rician noise). From the results over synthetic and real images we can conclude that this filter can be successfully used for automatic MR denoising. PMID- 18381248 TI - "Risk assessment of oral cancer in patients with pre-cancerous states of the oral cavity using micronucleus test and challenge assay" [Saran R, Tiwari RK, Reddy PP, Ahuja YR. Oral Oncol 2008;44:354-60]. PMID- 18381249 TI - A comparison of quantitative EEG features for neonatal seizure detection. AB - OBJECTIVE: This study was undertaken to identify the best performing quantitative EEG features for neonatal seizures detection from a test set of 21. METHODS: Each feature was evaluated on 1-min, artefact-free segments of seizure and non-seizure neonatal EEG recordings. The potential utility of each feature for neonatal seizure detection was determined using receiver operating characteristic analysis and repeated measures t-tests. A performance estimate of the feature set was obtained using a cross-fold validation and combining all features together into a linear discriminant classifier model. RESULTS: Significant differences between seizure and non-seizure segments were found in 19 features for 17 patients. The best performing features for this application were the RMS amplitude, the line length and the number of local maxima and minima. An estimate of the patient independent classifier performance yielded a sensitivity of 81.08% and specificity of 82.23%. CONCLUSIONS: The individual performances of 21 quantitative EEG features in detecting electrographic seizure in the neonate were compared and numerically quantified. Combining all features together into a classifier model led to superior performance than that provided by any individual feature taken alone. SIGNIFICANCE: The results documented in this study may provide a reference for the optimum quantitative EEG features to use in developing and enhancing neonatal seizure detection algorithms. PMID- 18381250 TI - Sensory gating in epilepsy - effects of the lateralization of hippocampal sclerosis. AB - OBJECTIVE: The role of the hippocampus in sensory gating is unresolved as yet. The current study was conducted in order to examine the effects of hippocampal lesions on the function of auditory sensory gating. METHODS: Epilepsy patients with unilateral and bilateral hippocampal sclerosis (HS) were investigated. Auditory-evoked potentials (AEPs) were recorded by multichannel scalp EEG in a double-click experiment. Gating was defined as the response decrease of the AEP components from 1st to 2nd click. RESULTS: Diagnosis (left vs. right vs. bilateral HS) did not affect the amplitudes or gating of the P50, N100 or P200 components. However, diagnosis had an impact on the topography of the N100 component after its peak maximum: In right HS patients, the N100 was left lateralized, while it was nearly symmetrically distributed in patients with left HS and right lateralized in patients with bilateral HS. Besides the N100, the topography of the P200 component was affected by diagnosis. CONCLUSIONS: Findings suggest that auditory activity, as reflected in the N100 and P200 components, is modulated by the hippocampus, but not sensory gating in its classical definition. SIGNIFICANCE: Deficits in P50 gating in schizophrenia are unlikely to be explained by hippocampal deficits. PMID- 18381251 TI - Facio-mandibular myoclonus specific during REM sleep. PMID- 18381252 TI - Embryonic expression of p68, a DEAD-box RNA helicase, in the oligochaete annelid Tubifex tubifex. AB - We have cloned and characterized the expression of a p68 homologue (designated Ttu-p68) from the oligochaete annelid Tubifex tubifex. Ttu-p68 mRNA is distributed broadly throughout the early stages. Ttu-p68 is expressed in all of the early blastomeres, in which Ttu-p68 RNA associates with pole plasms. Ttu-p68 transcripts are concentrated to 4d cell but not to 2d cell. During gastrulation, expression of Ttu-p68 is restricted to elongating germ bands (GBs) and an anteriormost crescent of micromere descendants on both sides of the embryo. During body elongation that follows gastrulation, expression of Ttu-p68 is further restricted to the stomodaeum (derived from the micromere crescent), ventral ganglia, lateral dots (corresponding to dorsal and ventral setal sacs), ventral large cells (that resemble presumptive primordial germ cells) in segments VIII-XII, and a bilateral pair of cell clusters at the caudal end. At the end of embryogenesis, Ttu-p68 expression persists exclusively in the tail and the lining epithelium of the pharynx. PMID- 18381253 TI - Bilirubin adsorption property of sol-gel-derived titania particles for blood purification therapy. AB - Titania (anatase) gel powders were prepared by peptizing commercially available titania sols and heating them at temperatures up to 700 degrees C, as candidates for bilirubin adsorbents for blood purification therapy. Those titania particles were in contact with a protein solution containing bilirubin and bovine serum albumin that mimics the blood of bilirubinemia patients. The amount of free or direct bilirubin in the solution insignificant. Indirect bilirubin or a bilirubin complex with albumin was adsorbed on the anatase powders, the primary particle size of which was as large as or larger than the size of an albumin molecule. The surface charge and surface charge density were only minor factors in controlling the indirect bilirubin adsorption. The present results indicated that the size of primary particles and hydrophobicity were significant for the sol-derived anatase in terms of bilirubin adsorption, and both were controllable by the heating temperature and the time period. PMID- 18381255 TI - Maximum strength for intermolecular adhesion of nanospheres at an optimal size. AB - Previous studies have emphasized that the adhesion strength between solid objects tends to increase as the characteristic size of the objects decreases and eventually saturates at the theoretical adhesion strength below a critical size scale. Here we show that the adhesion strength between two spheres or between a sphere and a solid half-space actually exhibits a peak value at an optimal size. This optimal size arises owing to a transition between surface- and bulk dominated interaction regimes at the nanoscale. PMID- 18381256 TI - Enhancement of chromatic contrast increases predation risk for striped butterflies. AB - Many prey species have evolved defensive colour patterns to avoid attacks. One type of camouflage, disruptive coloration, relies on contrasting patterns that hinder predators' ability to recognize an object. While high contrasts are used to facilitate detection in many visual communication systems, they are thought to provide misleading information about prey appearance in disruptive patterns. A fundamental tenet in disruptive coloration theory is the principle of 'maximum disruptive contrast', i.e. disruptive patterns are more effective when higher contrasts are involved. We tested this principle in highly contrasting stripes that have often been described as disruptive patterns. Varying the strength of chromatic contrast between stripes and adjacent pattern elements in artificial butterflies, we found a strong negative correlation between survival probability and chromatic contrast strength. We conclude that too high a contrast leads to increased conspicuousness rather than to effective camouflage. However, artificial butterflies that sported contrasts similar to those of the model species Limenitis camilla survived equally well as background-matching butterflies without these stripes. Contrasting stripes do thus not necessarily increase predation rates. This result may provide new insights into the design and characteristics of a range of colour patterns such as sexual, mimetic and aposematic signals. PMID- 18381254 TI - Optical tweezers for single cells. AB - Optical tweezers (OT) have emerged as an essential tool for manipulating single biological cells and performing sophisticated biophysical/biomechanical characterizations. Distinct advantages of using tweezers for these characterizations include non-contact force for cell manipulation, force resolution as accurate as 100aN and amiability to liquid medium environments. Their wide range of applications, such as transporting foreign materials into single cells, delivering cells to specific locations and sorting cells in microfluidic systems, are reviewed in this article. Recent developments of OT for nanomechanical characterization of various biological cells are discussed in terms of both their theoretical and experimental advancements. The future trends of employing OT in single cells, especially in stem cell delivery, tissue engineering and regenerative medicine, are prospected. More importantly, current limitations and future challenges of OT for these new paradigms are also highlighted in this review. PMID- 18381257 TI - Social networks in the lek-mating wire-tailed manakin (Pipra filicauda). AB - How social structure interacts with individual behaviour and fitness remains understudied despite its potential importance to the evolution of cooperation. Recent applications of network theory to social behaviour advance our understanding of the role of social interactions in various contexts. Here we applied network theory to the social system of lek-mating wire-tailed manakins (Pipra filicauda, Pipridae, Aves). We analysed the network of interactions among males in order to begin building a comparative framework to understand where coordinated display behaviour lies along the continuum from solitary to obligately cooperative dual-male displays in the family Pipridae. Network degree (the number of links from a male to others) ranged from 1 to 10, with low mean and high variance, consistent with the theory for the evolution of cooperation within social networks. We also assessed factors that could predict social and reproductive success of males. Four network metrics, degree, eigenvector centrality, information centrality and reach, some of which assess circuitous as well as the shortest (geodesic) paths of male connectivity, predicted male social rise. The duration of a male's territorial tenure during the 4 years of the study predicted his probability of siring offspring. PMID- 18381258 TI - Molecular anatomy of the cytoplasmic domain of bovine growth hormone receptor, a quantitative trait locus. AB - Quantitative trait loci (QTL) studies have indicated growth hormone receptor (GHR) as a candidate gene affecting cattle milk yield and composition. In order to characterize genetic variation at GHR in cattle, we studied European and East African breeds with different histories of selection, and Bos grunniens, Ovis aries, Sus scrofa, Bison bison and Rangifer tarandus as references. We sequenced most of the cytoplasmic domain (900 bp of exon 10), 89 bp of exon 8, including the putative causative mutation for the QTL effect, and 390 bp of intron 8 for comparison. In the cytoplasmic domain, seven synonymous and seven non-synonymous single nucleotide polymorphisms (SNP) were identified in cattle. Three non synonymous SNPs were found in sheep and one synonymous SNP in yak, while other studied species were monomorphic. Three major haplotypes were observed, one unique to African breeds, one unique to European breeds and one shared. Bison and yak haplotypes are derivatives of the European haplotype lineage. Most of the exon 10 non-synonymous cattle SNPs appear at phylogenetically highly conserved sites. The polymorphisms in exon 10 cluster around a ruminant-specific tyrosine residue, suggesting that this site may act as an additional signalling domain of GHR in ruminants. Alternative explanations for the persistent polymorphism include balancing selection, hitch-hiking, pleiotropic or sexually antagonistic fitness effects or relaxed functional constraints. PMID- 18381260 TI - Does habituation to humans influence predator discrimination in Gunther's dik diks (Madoqua guentheri)? AB - Animals living around humans may habituate to us, but little is known about the consequences of this habituation. Some wildlife managers assume that habituation to humans makes individuals less likely to respond to natural predators, which is something to be avoided in captive breeding programmes where animals are destined for release. We conducted a playback experiment where we broadcast the sounds of a terrestrial predator and the song from a non-threatening bird to Gunther's dik diks (Madoqua guentheri), a small ungulate that is vulnerable to many predators, in areas where dik-diks were and were not habituated to humans. Contrary to our expectation, habituated dik-diks discriminated the predator sounds from the birdsong, while unhabituated dik-diks failed to make this discrimination. Our results demonstrate that humans may influence predation hazard assessment, but we should not generally assume that human-habituated animals will be especially vulnerable to predators. PMID- 18381259 TI - Anisotropic patterned population synchrony in climatic gradients indicates nonlinear climatic forcing. AB - Although climatic forcing has been suspected to be the most common cause of spatial population synchrony owing to the Moran effect, it has proved difficult to disentangle the impact of climate from other possible causes of synchrony based on population survey data. Nonlinear population responses to climatic variation may be a part of this difficulty, but they can also provide an opportunity to highlight the climate impacts through targeted survey designs. In particular, when species distribution ranges encompass consistent spatial gradients in climate (e.g. according to latitude or altitude), such gradients can be strategically included in the spatial design of population surveys as to facilitate comparisons of spatial synchrony patterns across and along the gradient. In that case, we predict that nonlinear impacts of climatic variation on population growth rates will result in anisotropic (direction specific) synchrony patterns in the sense that synchrony will drop faster with distance along the climatic gradient than across it. We provide an empirical case study to exemplify survey design and analyses. Of two sympatric species of geometrids, inhabiting an altitudinal gradient in subarctic birch forest, one (Operophtera brumata L.) showed anisotropic synchrony consistent with a strongly nonlinear sensitivity to climatic variation, whereas the other (Epirrita autumnata Bkh.) did not. These results are interpreted in light of the biological characteristics of the species. PMID- 18381261 TI - Long-term persistence of GM oilseed rape in the seedbank. AB - Coexistence between genetically modified (GM) and non-GM plants is a field of rapid development and considerable controversy. In crops, it is increasingly important to understand and predict the GM volunteer emergence in subsequent non GM crops. Theoretical models suggest recruitment from the seedbank over extended periods, but empirical evidence matching these predictions has been scarce. Here, we provide evidence of long-term GM seed persistence in conventional agriculture. Ten years after a trial of GM herbicide-tolerant oilseed rape, emergent seedlings were collected and tested for herbicide tolerance. Seedlings that survived the glufosinate herbicide (15 out of 38 volunteers) tested positive for at least one GM insert. The resulting density was equivalent to 0.01 plants m-2, despite complying with volunteer reduction recommendations. These results are important in relation to debating and regulating coexistence of GM and non-GM crops, particularly for planting non-GM crops after GM crops in the same field. PMID- 18381262 TI - Is breeding of farmland wading birds depressed by a combination of predator abundance and grazing? AB - Agri-environment schemes have been implemented across Europe to counter biodiversity loss in agricultural landscapes and halt the continual decline of farmland birds, including waders. Such schemes provide financial compensation for changes in agricultural practice, including livestock grazing regimes. Scheme uptake has been variable, partly because farmers believe that other factors, notably predation, are key to wader population declines. On the basis of wader breeding surveys across Shetland, UK, we show that predator density and livestock grazing, through reducing sward height, interact to influence territoriality and thereby are likely to affect wader breeding success. Our results appear to reflect views of both farmers and government agencies, which indicates that future agri-environment schemes would benefit from genuine stakeholder participation to maximize scheme uptake, implementation and beneficial effects on biodiversity. Our findings also imply that agri-environment schemes will reap the greatest benefits for waders through reducing stocking rate where avian predators are abundant. PMID- 18381263 TI - Effects of mutual shading of tree crowns on prediction of photosynthetic light use efficiency in a coastal Douglas-fir forest. AB - Gross primary production (GPP) is often expressed as the product of absorbed photosynthetically active radiation and the efficiency (epsilon) with which a plant community uses absorbed radiation in biomass production. Light-use efficiency is affected by environmental stresses, and varies diurnally and seasonally. Uncertainty about epsilon can be a serious limitation when modeling GPP. An important determinant of epsilon is the amount and type of solar radiation incident on a canopy, because an abundance of light can trigger a photo protective reaction, diminishing GPP. The radiation regime in a forest canopy is determined by the predominant sky conditions and by mutual shading of tree crowns. Shading effects, producing shifts in the amount of incident direct and diffuse solar radiation, have been largely ignored, however, because they depend on forest structure and are difficult to measure. We describe a new approach for estimating changes in mutual canopy shading throughout the day and year based on a canopy structure model derived from light detection and ranging (LiDAR). Proportions of canopy shading were then combined with eddy covariance data to assess the explanatory power for variance in epsilon by regression tree analysis over half-hourly, daily and weekly time scales. The approach explained between 75 and 97% of variance in epsilon, representing an increase of between 5 and 16% compared with models driven solely by meteorological variables. PMID- 18381264 TI - Biomechanical differences in the stem straightening process among Pinus pinaster provenances. A new approach for early selection of stem straightness. AB - Stem straightness is an important selection trait in Pinus pinaster Ait. breeding programs. Despite the stability of stem straightness rankings in provenance trials, the efficiency of breeding programs based on a quantitative index of stem straightness remains low. An alternative approach is to analyze biomechanical processes that underlie stem form. The rationale for this selection method is that genetic differences in the biomechanical processes that maintain stem straightness in young plants will continue to control stem form throughout the life of the tree. We analyzed the components contributing most to genetic differences among provenances in stem straightening processes by kinetic analysis and with a biomechanical model defining the interactions between the variables involved (Fournier's model). This framework was tested on three P. pinaster provenances differing in adult stem straightness and growth. One-year-old plants were tilted at 45 degrees, and individual stem positions and sizes were recorded weekly for 5 months. We measured the radial extension of reaction wood and the anatomical features of wood cells in serial stem cross sections. The integral effect of reaction wood on stem leaning was computed with Fournier's model. Responses driven by both primary and secondary growth were involved in the stem straightening process, but secondary-growth-driven responses accounted for most differences among provenances. Plants from the straight-stemmed provenance showed a greater capacity for stem straightening than plants from the sinuous provenances mainly because of (1) more efficient reaction wood (higher maturation strains) and (2) more pronounced secondary-growth-driven autotropic decurving. These two process-based traits are thus good candidates for early selection of stem straightness, but additional tests on a greater number of genotypes over a longer period are required. PMID- 18381265 TI - Resin flow responses to fertilization, wounding and fungal inoculation in loblolly pine (Pinus taeda) in North Carolina. AB - Resin flow is the primary means of natural defense against southern pine beetle (Dendroctonus frontalis Zimm.), the most important insect pest of Pinus spp. in the southern United States. As a result, factors affecting resin flow are of interest to researchers and forest managers. We examined the influence of fertilization, artificial wounding and fungal inoculation on resin flow in 6- and 12-year-old stands of loblolly pine (Pinus taeda L.) and determined the extent of that influence within and above the wounded stem area and through time. Fertilization increased constitutive resin flow, but only the younger trees sustained increased resin flow after wounding and inoculation treatments. An induced resin flow response occurred between 1 and 30 days after wounding and inoculation treatments. Wounding with inoculation resulted in greater resin flow than wounding alone, but increasing amounts of inoculum did not increase resin flow. Increased resin flow (relative to controls) lasted for at least 90 days after wounding and inoculation. This increase appeared to be limited to the area of treatment, at least in younger trees. The long-lasting effects of fungal inoculation on resin flow, as well as the response to fertilization, suggest that acquired resistance through induced resin flow aids in decreasing susceptibility of loblolly pine to southern pine beetle. PMID- 18381266 TI - Calcium addition at the Hubbard Brook Experimental Forest increases sugar storage, antioxidant activity and cold tolerance in native red spruce (Picea rubens). AB - In fall (November 2005) and winter (February 2006), we collected current-year foliage of native red spruce (Picea rubens Sarg.) growing in a reference watershed and in a watershed treated in 1999 with wollastonite (CaSiO(3), a slow release calcium source) to simulate preindustrial soil calcium concentrations (Ca addition watershed) at the Hubbard Brook Experimental Forest (Thornton, NH). We analyzed nutrition, soluble sugar concentrations, ascorbate peroxidase (APX) activity and cold tolerance, to evaluate the basis of recent (2003) differences between watersheds in red spruce foliar winter injury. Foliar Ca and total sugar concentrations were significantly higher in trees in the Ca-addition watershed than in trees in the reference watershed during both fall (P=0.037 and 0.035, respectively) and winter (P=0.055 and 0.036, respectively). The Ca-addition treatment significantly increased foliar fructose and glucose concentrations in November (P=0.013 and 0.007, respectively) and foliar sucrose concentrations in winter (P=0.040). Foliar APX activity was similar in trees in both watersheds during fall (P=0.28), but higher in trees in the Ca-addition watershed during winter (P=0.063). Cold tolerance of foliage was significantly greater in trees in the Ca-addition watershed than in trees in the reference watershed (P<0.001). Our results suggest that low foliar sugar concentrations and APX activity, and reduced cold tolerance in trees in the reference watershed contributed to their high vulnerability to winter injury in 2003. Because the reference watershed reflects forest conditions in the region, the consequences of impaired physiological function caused by soil Ca depletion may have widespread implications for forest health. PMID- 18381267 TI - Cambial phenology, wood formation and temperature thresholds in two contrasting years at high altitude in southern Italy. AB - Xylogenesis was monitored during 2003 and 2004 in a timberline environment in southern Italy to assess links between temperature, cambial phenology and wood formation on a short-time scale. Wood microcores were collected weekly from May to October from 10 trees of Pinus leucodermis Ant., histological sections were cut with a rotary microtome and anatomical features of the developing and mature tracheids were observed and measured along the growing tree ring. Spring 2003 was hotter than spring 2004, with temperatures up to 2.6 degrees C above historical means. The hotter conditions resulted in an earlier onset of cambial activity and all differentiation phases of about 20 days, resulting in an increased duration of xylogenesis of about 23 days. Air and stem temperatures at which xylogenesis had a 0.5 probability of being active were calculated with logistic regressions fitted on binary responses. In both years, similar thresholds were estimated with daily mean values of 8.2 and 9.5 degrees C for air and stem temperatures, respectively. The observed convergent responses of cambium phenology to temperature during the two contrasting springs confirm the key role of this environmental factor in determining the onset and duration of wood formation in timberline areas. The intra-annual dynamics of ring-width increase differed between years, with significantly narrower rings formed in 2004 than in 2003. These differences were mainly related to cell size because larger earlywood tracheids were produced in 2003. This study demonstrates the plasticity of tree ring formation in response to high temperatures as a result of modifications in the onset and duration of differentiation. PMID- 18381268 TI - Interactions of water stress and solar irradiance on the physiology and biochemistry of Ligustrum vulgare. AB - We studied the interactive effects of water stress and solar irradiance on physiological and biochemical traits in Ligustrum vulgare L., with special emphasis on antioxidant enzymes and flavonoids. Water relations, photosynthetic performance, plant growth, activities of antioxidant enzymes and of phenylalanine ammonia-lyase, and concentrations of nonstructural carbohydrates and phenylpropanoids were measured in plants growing in 12% (shade) or 100% (sun) sunlight and supplied with 100 or 40% of daily evapotranspiration-demand over a 4 week period. The mild water stress treatment caused leaf water potential and relative water content to decline on average by -0.22 MPa and 4.5%, respectively. In response to the water stress treatment, photosynthetic rates decreased more in sun plants than in shade plants, likely because of declines in photosystem II photochemistry, apparent maximum rate of carboxylation and apparent maximum electron transport rate coupled with significant reductions in stomatal conductance. Antioxidant enzymatic activities, which were much greater in sun leaves than in shade leaves under well-watered conditions, increased (particularly the enzymatic activities associated with hydrogen peroxide removal) in response to water stress only in shade leaves. Antioxidant phenylpropanoids, particularly quercetin and luteolin derivatives, markedly increased in response to full sunlight irrespective of water treatment; however, antioxidant phenylpropanoid concentrations increased in response to water stress only in shade leaves. We suggest that: (1) assimilated carbon in sun plants was used largely to support an effective antioxidant system capable of countering water stress-induced oxidative damage--an example of cross tolerance; and (2) in shade plants, carbon was also diverted from growth to counter oxidative damage driven by the mild water-stress treatment. Both findings are consistent with the nearly exclusive distribution of L. vulgare in well-watered, partially shaded Mediterranean areas. PMID- 18381269 TI - Comparative analysis of transcript abundance in Pinus sylvestris after challenge with a saprotrophic, pathogenic or mutualistic fungus. AB - To investigate functional differences in the recognition and response mechanisms of conifer roots to fungi with different trophic strategies, Pinus sylvestris L. was challenged with a saprotrophic fungus Trichoderma aureoviride Rifai. The results were compared with separate studies investigating pine interactions with a pathogen, Heterobasidion annosum (Fr.) Bref. sensu stricto and an ectomycorrhizal symbiont, Laccaria bicolor Maire (Orton). Global changes in the expression of 2109 conifer genes were assayed 1, 5 and 15 days after inoculation. Gene expression data from a cDNA microarray were analyzed by the 2-interconnected mixed linear model statistical approach. The total number of genes differentially expressed compared with the uninfected control was similar after challenge with the pathogen and the ectomycorrhizal symbiont, but the number of differentially expressed genes increased over time for H. annosum, and decreased for L. bicolor. Inoculation of pine roots with T. aureoviride resulted overall in a much lower number of genes with changed transcript levels compared with inoculation with H. annosum or L. bicolor. Functional classification of the differentially expressed genes revealed that the ectomycorrhizal fungus triggered transient induction of defence-related genes. The response and induction of defence against the pathogen was delayed and the magnitude increased over time. Thus, there were specific transcriptional responses depending on whether the conifer roots were challenged with mutualistic, saprotrophic or pathogenic fungi. This suggests that pine trees are able to recognize diverse fungal species and specifically distinguish whether they are pathogenic, neutral or beneficial microbial agents. PMID- 18381270 TI - Molecular cloning of a putative gene encoding isopentenyltransferase from pingyitiancha (Malus hupehensis) and characterization of its response to nitrate. AB - A putative isopentenyltransferase (IPT) encoding gene was identified from a pingyitiancha (Malus hupehensis Rehd.) expressed sequence tag database, and the full-length gene was cloned by RACE. Based on expression profile and sequence alignment, the nucleotide sequence of the clone, named MhIPT3, was most similar to AtIPT3, an IPT gene in Arabidopsis. The full-length cDNA contained a 963-bp open reading frame encoding a protein of 321 amino acids with a molecular mass of 37.3 kDa. Sequence analysis of genomic DNA revealed the absence of introns in the frame. Quantitative real-time PCR analysis demonstrated that the gene was expressed in roots, stems and leaves. Application of nitrate to roots of nitrogen deprived seedlings strongly induced expression of MhIPT3 and was accompanied by the accumulation of cytokinins, whereas MhIPT3 expression was little affected by ammonium application to roots of nitrogen-deprived seedlings. Application of nitrate to leaves also up-regulated the expression of MhIPT3 and corresponded closely with the accumulation of isopentyladenine and isopentyladenosine in leaves. PMID- 18381271 TI - SAGE transcript profiling of the juvenile cambial region of Eucalyptus grandis. AB - Despite the importance of Eucalyptus spp. in the pulp and paper industry, functional genomic approaches have only recently been applied to understand wood formation in this genus. We attempted to establish a global view of gene expression in the juvenile cambial region of Eucalyptus grandis Hill ex Maiden. The expression profile was obtained from serial analysis of gene expression (SAGE) library data produced from 3- and 6-year-old trees. Fourteen-base expressed sequence tags (ESTs) were searched against public Eucalyptus ESTs and annotated with GenBank. Altogether 43,304 tags were generated producing 3066 unigenes with three or more copies each, 445 with a putative identity, 215 with unknown function and 2406 without an EST match. The expression profile of the juvenile cambial region revealed the presence of highly frequent transcripts related to general metabolism and energy metabolism, cellular processes, transport, structural components and information pathways. We made a quantitative analysis of a large number of genes involved in the biosynthesis of cellulose, pectin, hemicellulose and lignin. Our findings provide insight into the expression of functionally related genes involved in juvenile wood formation in young fast-growing E. grandis trees. PMID- 18381272 TI - Molecular cloning and expression analysis of a gene encoding a putative beta ketoacyl-acyl carrier protein (ACP) synthase III (KAS III) from Jatropha curcas. AB - A cDNA clone encoding a putative beta-ketoacyl-acyl carrier protein (ACP) synthase III (KAS III) was isolated from Jatropha curcas L., a woody oil plant. The cDNA clone (named JcKAS III) contained a 1203-bp open reading frame coding for 400 amino acids with a predicted molecular mass of about 42 kDa. The deduced amino acid sequence of the cDNA clone shares about 80% identity to KAS III from other plants, and contains a conserved Cys(176) in the active site and the amino acid motif G(355)NTSAAS(361) which is responsible for binding regulatory acyl ACPs. Southern blotting analysis indicated that JcKAS III is a single copy gene in the J. curcas genome. Quantitative real-time PCR analysis showed that JcKAS III was expressed in all tissues examined with highest expression in roots, and that expression of JcKAS III increased as seeds developed. PMID- 18381273 TI - A MADS-box gene of Populus deltoides expressed during flower development and in vegetative organs. AB - A MADS-box gene (PdPI) was isolated from a cDNA library constructed from male flower buds of Populus deltoides Bartr. ex Marsh. According to an analysis of genomic DNA structure and putative protein structure, and a phylogenetic study, PdPI is an ortholog of the Arabidopsis PI gene. Relative-quantitative real-time polymerase chain reaction analysis showed that PdPI has a broader expression pattern than PI in Arabidopsis. PdPI was strongly expressed in floral buds and roots and weakly expressed in immature xylem, leaves and apical buds of the male P. deltoides tree. In male inflorescences, PdPI expression was abundant in the perianth and anther, and weak in the peduncle and mature pollen. The large differences in PdPI expression at various phases of male floral bud development were closely related to the development of flower organs (perianth and stamen) and pollen. PdPI was also expressed in female inflorescences. Our results suggest that PdPI has multiple functions in the development of P. deltoides. PMID- 18381274 TI - Acclimation of tropical tree species to hurricane disturbance: ontogenetic differences. AB - We investigated acclimation responses of seedlings and saplings of the pioneer species Cecropia schreberiana Miq. and three non-pioneer species, Dacryodes excelsa Vahl, Prestoea acuminata (Willdenow) H.E. Moore var. montana (Graham) Henderson and Galeano, and Sloanea berteriana Choisy ex DC, following a hurricane disturbance in a lower montane wet forest in Puerto Rico. Measurements were made, shortly after passage of the hurricane, on leaves expanded before the hurricane (pre-hurricane leaves) and, at a later time, on recently matured leaves that developed after the hurricane (post-hurricane leaves) from both seedlings and saplings at sites that were severely damaged by the hurricane (disturbed sites) and at sites with little disturbance (undisturbed sites). Pre-hurricane leaves of the non-pioneer species had relatively low light-saturated photosynthetic rates (A(max)) and stomatal conductance (g(s)); neither A(max) nor g(s) responded greatly to the increase in irradiance that resulted from the disturbance, and there were few significant differences between seedlings and saplings. Pre hurricane leaves of plants at undisturbed sites had low dark respiration rates per unit area (R(d)) and light compensation points (LCP), whereas pre-hurricane leaves of plants at disturbed sites had significantly higher R(d) and LCP. Post hurricane leaves of plants at disturbed sites had significantly higher A(max) and R(d) than plants at undisturbed sites. Compared with seedlings, saplings had higher A(max) and R(d) and showed greater acclimation to the increase in irradiance that followed the disturbance. Post-hurricane leaves of the non pioneer species had significantly lower A(max) and were less responsive to changes in irradiance than the pioneer species C. schreberiana. Variation in A(max) across light environments and stages was strongly related to differences in leaf mass per unit area (LMA), especially in the non-pioneer species. As indicated by V(cmax) or J(max) per unit nitrogen, light acclimation of A(max) was determined by leaf morphology (LMA) for the non-pioneer species and by both leaf morphology and leaf biochemistry for C. schreberiana. Ontogenetic changes in A(max) were attributable to changes in leaf morphology. The ontogenetic component of variation in A(max) across light environments and stages differed among species, ranging from 36 to 59% for the non-pioneer species (D. excelsa, 59.3%; P. acuminata var. montana, 44.7%; and S. berteriana, 36.3%) compared with only 17% in the pioneer species C. schreberiana. PMID- 18381275 TI - Ionic homeostasis and reactive oxygen species control in leaves and xylem sap of two poplars subjected to NaCl stress. AB - We investigated the effects of increasing soil NaCl concentration on intracellular compartmentalization of salt and on the activities of antioxidant enzymes (superoxide dismutase (SOD), ascorbic peroxidase (APX), catalase (CAT) and glutathione reductase (GR)) and their role in the regulation of reactive oxygen species (ROS; O(2)(-*) and H(2)O(2)) in leaves and xylem sap of salt tolerant Populus euphratica Oliv. and salt-sensitive P. popularis cv. 35-44. Mesophyll cells of P. euphratica exhibited a high capacity for NaCl exclusion and compartmentalization of salt in vacuoles compared with P. popularis. In P. popularis, the salt treatment resulted in large accumulations of Na(+) and Cl(-) in leaves that induced significant increases in O(2)(-*) and H(2)O(2) production despite marked increases in the activities of antioxidant enzymes in leaves and xylem sap. Separation of the isoforms of leaf SOD, APX and CAT by polyacrylamide gel electrophoresis followed by in-gel activity staining revealed that the salt induced activities of APX and CAT were the result of increases in activities of all the isoenzymes. Leaf injury and shedding of aged leaves occurred following the oxidative burst in P. popularis, indicating that the increased activities of antioxidant enzymes in P. popularis were insufficient to counter the harmful effects of ROS at high soil NaCl concentrations. Unlike P. popularis plants, P. euphratica plants did not exhibit an oxidative burst in response to the NaCl treatments, because of (1) a high salt exclusion capacity and effective compartmentalization of salt in vacuoles, and (2) up-regulation of antioxidant enzymatic activities after the onset of salt stress. We conclude that P. euphratica plants subjected to saline conditions control ROS homeostasis through two pathways: (1) by maintaining cellular ionic homeostasis and thereby limiting the NaCl-induced enhancement of ROS production under long-term saline conditions; and (2) by rapidly up-regulating antioxidant defenses to prevent oxidative damage. PMID- 18381276 TI - Comparative measurements of transpiration and canopy conductance in two mixed deciduous woodlands differing in structure and species composition. AB - Transpiration of two heterogeneous broad-leaved woodlands in southern England was monitored by the sap flux technique throughout the 2006 growing season. Grimsbury Wood, which had a leaf area index (LAI) of 3.9, was dominated by oak (Quercus robur L.) and birch (Betula pubescens L.) and had a continuous hazel (Corylus avellana L.) understory. Wytham Woods, which had an LAI of 3.6, was dominated by ash (Fraxinus excelsior L.) and sycamore (Acer pseudoplatanus L.) and had only a sparse understory. Annual canopy transpiration was 367 mm for Grimsbury Wood and 397 mm for Wytham Woods. These values were similar to those for beech (Fagus sylvatica L.) plantations in the same region, and differ from one another by less than the typical margin of uncertainty of the sap flux technique. Canopy conductance (g(c)), calculated for both woodlands by inverting the Penman Monteith equation, was related to incoming solar radiation (R(G)) and the vapor pressure deficit (D). The response of g(c) to R(G) was similar for both forests. Both reference conductance (g(cref)), defined as g(c) at D=1 kPa, and stomatal sensitivity (-m), defined as the slope of the logarithmic response curve of g(c) to D, increased during the growing season at Wytham Woods but not at Grimsbury Wood. The -m/g(cref) ratio was significantly lower at Wytham Woods than at Grimsbury Wood and was insufficient to keep the difference between leaf and soil water potentials constant, according to a simple hydraulic model. This meant that annual water consumption of the two woodlands was similar despite different regulatory mechanisms and associated short-term variations in canopy transpiration. The -m/g(cref) ratio depended on the range of D under which the measurements were made. This was shown to be particularly important for studies conducted under low and narrow ranges of D. PMID- 18381277 TI - Water relations and drought-induced embolism in olive (Olea europaea) varieties 'Meski' and 'Chemlali' during severe drought. AB - We examined the effects of drought on the water relations, osmotic adjustment and xylem vulnerability to embolism of olive (Olea europaea L.) varieties, 'Meski' and 'Chemlali'. Two-year-old self-rooted cuttings growing in sand-filled pots in a greenhouse were subjected to water stress by withholding water for 60 days. Water relations and gas exchange measurements showed that 'Chemlali' was more drought resistant than 'Meski' and had a greater capacity for osmotic adjustment through solute accumulation. However, when water stress was acute, the effect of osmoregulation on leaf cell turgor was largely counteracted by xylem cavitation. Cavitation vulnerability curves showed that both varieties were highly resistant to embolism formation. The xylem water potential inducing 50% loss of stem conductivity approached -7 MPa in 'Meski' and only slightly less in 'Chemlali'. Although the difference between varieties in susceptibility to xylem embolism was small, it appears to account in large part for the difference between them in the ability to tolerate severe drought. PMID- 18381278 TI - Intra-plant variation in cyanogenesis and the continuum of foliar plant defense traits in the rainforest tree Ryparosa kurrangii (Achariaceae). AB - At the intra-plant level, temporal and spatial variations in plant defense traits can be influenced by resource requirements, defensive priorities and storage opportunities. Across a leaf age gradient, cyanogenic glycoside concentrations in the rainforest understory tree Ryparosa kurrangii B.L. Webber were higher in young expanding leaves than in mature leaves (2.58 and 1.38 mg g(-1), respectively). Moreover, cyanogens, as an effective chemical defense against generalist herbivores, contributed to a defense continuum protecting foliar tissue during leaf development. Chemical (cyanogens and phenolic compounds) and phenological (delayed greening) defense traits protected young leaves, whereas mature leaves were largely protected by physical defense mechanisms (lamina toughness; explained primarily by leaf mass per area). Cyanogen concentration was considerably higher in floral tissue than in foliar tissue and decreased in floral tissue during development. Across contrasting tropical seasons, foliar cyanogenic concentration varied significantly, being highest in the late wet season and lowest during the pre-wet season, the latter coinciding with fruiting and leaf flushing. Cyanogens in R. kurrangii appear to be differentially allocated in a way that maximizes plant fitness but may also act as a store of reduced nitrogen that is remobilized during flowering and leaf flushing. PMID- 18381279 TI - Combinatorial modification of human histone H4 quantitated by two-dimensional liquid chromatography coupled with top down mass spectrometry. AB - Quantitative proteomics has focused heavily on correlating protein abundances, ratios, and dynamics by developing methods that are protein expression-centric (e.g. isotope coded affinity tag, isobaric tag for relative and absolute quantification, etc.). These methods effectively detect changes in protein abundance but fail to provide a comprehensive perspective of the diversity of proteins such as histones, which are regulated by post-translational modifications. Here, we report the characterization of modified forms of HeLa cell histone H4 with a dynamic range >10(4) using a strictly Top Down mass spectrometric approach coupled with two dimensions of liquid chromatography. This enhanced dynamic range enabled the precise characterization and quantitation of 42 forms uniquely modified by combinations of methylation and acetylation, including those with trimethylated Lys-20, monomethylated Arg-3, and the novel dimethylated Arg-3 (each <1% of all H4 forms). Quantitative analyses revealed distinct trends in acetylation site occupancy depending on Lys-20 methylation state. Because both modifications are dynamically regulated through the cell cycle, we simultaneously investigated acetylation and methylation kinetics through three cell cycle phases and used these data to statistically assess the robustness of our quantitative analysis. This work represents the most comprehensive analysis of histone H4 forms present in human cells reported to date. PMID- 18381280 TI - Functional surfaces on the p35/ARPC2 subunit of Arp2/3 complex required for cell growth, actin nucleation, and endocytosis. AB - The Arp2/3 complex is comprised of seven evolutionarily conserved subunits and upon activation by WASp or another nucleation promoting factor nucleates the formation of actin filaments. These events are critical for driving a wide range of cellular processes, including motility, endocytosis, and intracellular trafficking. However, an in depth understanding of the Arp2/3 complex activation and nucleation mechanism is still lacking. Here, we used a mutagenesis approach in Saccharomyces cerevisiae to dissect the structural and functional roles of the p35/ARPC2 subunit. Using integrated alleles that target conserved and solvent exposed residues, we identified surfaces on p35/ARPC2 required for cell growth, actin organization, and endocytosis. In parallel, we purified the mutant Arp2/3 complexes and compared their actin assembly activities both in the presence and in the absence of WASp. The majority of alleles with defects mapped to one face of p35/ARPC2, where there was a close correlation between loss of actin nucleation and endocytosis. A second site required for nucleation and endocytosis was identified near the contact surface between p35/ARPC2 and p19/ARPC4. A third site was identified at a more distal conserved surface, which was critical for endocytosis but not nucleation. These findings pinpoint the key surfaces on p35/ARPC2 required for Arp2/3 complex-mediated actin assembly and cellular function and provide a higher resolution view of Arp2/3 structure and mechanism. PMID- 18381281 TI - Naturally occurring disulfide-bound dimers of three-fingered toxins: a paradigm for biological activity diversification. AB - Disulfide-bound dimers of three-fingered toxins have been discovered in the Naja kaouthia cobra venom; that is, the homodimer of alpha-cobratoxin (a long-chain alpha-neurotoxin) and heterodimers formed by alpha-cobratoxin with different cytotoxins. According to circular dichroism measurements, toxins in dimers retain in general their three-fingered folding. The functionally important disulfide 26 30 in polypeptide loop II of alpha-cobratoxin moiety remains intact in both types of dimers. Biological activity studies showed that cytotoxins within dimers completely lose their cytotoxicity. However, the dimers retain most of the alpha cobratoxin capacity to compete with alpha-bungarotoxin for binding to Torpedo and alpha7 nicotinic acetylcholine receptors (nAChRs) as well as to Lymnea stagnalis acetylcholine-binding protein. Electrophysiological experiments on neuronal nAChRs expressed in Xenopus oocytes have shown that alpha-cobratoxin dimer not only interacts with alpha7 nAChR but, in contrast to alpha-cobratoxin monomer, also blocks alpha3beta2 nAChR. In the latter activity it resembles kappa bungarotoxin, a dimer with no disulfides between monomers. These results demonstrate that dimerization is essential for the interaction of three-fingered neurotoxins with heteromeric alpha3beta2 nAChRs. PMID- 18381282 TI - Appearance and propagation of polyglutamine-based amyloids in yeast: tyrosine residues enable polymer fragmentation. AB - In yeast, fragmentation of amyloid polymers by the Hsp104 chaperone allows them to propagate as prions. The prion-forming domain of the yeast Sup35 protein is rich in glutamine, asparagine, tyrosine, and glycine residues, which may define its prion properties. Long polyglutamine stretches can also drive amyloid polymerization in yeast, but these polymers are unable to propagate because of poor fragmentation and exist through constant seeding with the Rnq1 prion polymers. We proposed that fragmentation of polyglutamine amyloids may be improved by incorporation of hydrophobic amino acid residues into polyglutamine stretches. To investigate this, we constructed sets of polyglutamine with or without tyrosine stretches fused to the non-prion domains of Sup35. Polymerization of these chimeras started rapidly, and its efficiency increased with stretch size. Polymerization of proteins with polyglutamine stretches shorter than 70 residues required Rnq1 prion seeds. Proteins with longer stretches polymerized independently of Rnq1 and thus could propagate. The presence of tyrosines within polyglutamine stretches dramatically enhanced polymer fragmentation and allowed polymer propagation in the absence of Rnq1 and, in some cases, of Hsp104. PMID- 18381283 TI - Foxa2 activity increases plasma high density lipoprotein levels by regulating apolipoprotein M. AB - Obesity, diabetes, insulin resistance, and hyperinsulinemia are frequently associated with a cluster of closely related lipid abnormalities such as low plasma levels of high density lipoprotein (HDL) and elevated levels of triglyceride, both known to increase the risk of developing atherosclerotic disease. The molecular mechanisms linking obesity, insulin resistance, and hyperinsulinemia to low HDL levels are incompletely understood. Here we demonstrate that insulin, through a Foxa2-mediated mechanism, inhibited the expression of apolipoprotein M (apoM), an important determinant of plasma pre beta-HDL and alpha-HDL concentrations. Obese mice had decreased apoM expression and plasma pre-beta-HDL levels due to inactivation of Foxa2 in hyperinsulinemic states. Nuclear reexpression of Foxa2 with a phosphorylation-deficient mutant Foxa2T156A (Ad-T156A) activated apoM expression and increased plasma pre-beta-HDL and alpha-HDL levels. In contrast, haploinsufficient Foxa2(+/-) mice exhibited decreased hepatic apoM expression and plasma pre-beta-HDL and HDL levels. The increase in plasma HDL levels and pre-beta-HDL formation by Foxa2 was mediated exclusively by apoM, as constitutive active expression of Foxa2 in apoM(-/-) mice had no effect on plasma HDL levels. Our results identify a fundamental mechanism by which insulin regulates plasma HDL levels in physiological and insulin resistant states and thus have important implications for novel therapeutic approaches to prevent atherosclerosis. PMID- 18381285 TI - Permeation properties of a P2X receptor in the green algae Ostreococcus tauri. AB - We have cloned a P2X receptor (OtP2X) from the green algae Ostreococcus tauri. The 42-kDa receptor shares approximately 28% identity with human P2X receptors and 23% with the Dictyostelium P2X receptor. ATP application evoked flickery single channel openings in outside-out membrane patches from human embryonic kidney 293 cells expressing OtP2X. Whole-cell recordings showed concentration dependent cation currents reversing close to zero mV; ATP gave a half-maximal current at 250 mum. alphabeta-Methylene-ATP evoked only small currents in comparison to ATP (EC(50) > 5 mm). 2',3'-O-(4-Benzoylbenzoyl)-ATP, betagamma imido-ATP, ADP, and several other nucleotide triphosphates did not activate any current. The currents evoked by 300 mum ATP were not inhibited by 100 microm suramin, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid, 2',3'-O-(2,4,6 trinitrophenol)-ATP, or copper. Ion substitution experiments indicated permeabilities relative to sodium with the rank order calcium >choline >Tris >tetraethylammonium >N-methyl-D-glucosamine. However, OtP2X had a low relative calcium permeability (P(Ca)/P(Na) = 0.4) in comparison with other P2X receptors. This was due at least in part to the presence of an asparagine residue (Asn(353)) at a position in the second transmembrane domain in place of the aspartate that is completely conserved in all other P2X receptor subunits, because replacement of Asn(353) with aspartate increased calcium permeability by approximately 50%. The results indicate that the ability of ATP to gate cation permeation across membranes exists in cells that diverged in evolutionary terms from animals about 1 billion years ago. PMID- 18381286 TI - The substrates of the gamma-aminobutyric acid transporter GAT-1 induce structural rearrangements around the interface of transmembrane domains 1 and 6. AB - The sodium- and chloride-coupled gamma-aminobutyric acid (GABA) transporter GAT-1 is essential for efficient synaptic transmission by this neurotransmitter. GAT-1 is the first cloned member of the neurotransmitter-sodium-symporter family. Here we address the idea that during transport the extracellular halves of transmembrane domains (TM) 1 and 6, TM 1b/TM 6a, move relative to the binding pocket. Therefore, we have probed the aqueous accessibility of TM 6a and its proximity to TM 1b in the presence and absence of its substrates. Cysteines were introduced, one by one, at all TM 6a positions. In several mutants, transport activity was inhibited by the impermeant sulfhydryl reagent (2 trimethylammonium)methanethiosulfonate, whereas wild type GAT-1 was basically insensitive. This inhibition was potentiated by sodium, whereas GABA was protective. Moreover, we used paired cysteine mutagenesis in conjunction with treatments with copper(II)(1,10-phenanthroline)(3) (CuPh). CuPh did not affect the activity of wild type GAT-1 but potently inhibited transport by the TM 6a mutant D287C. Such inhibition was not observed with D287C/C74A, indicating that Asp-287 is close to Cys-74 of TM 1b. Inhibition of transport of D287C by CuPh, but not by (2-trimethylammonium)methanethiosulfonate, was potentiated when sodium and GABA were both removed. Thus, the degree of inhibition by CuPh is not a simple function of the accessibility of the individual cysteines but also involves structural rearrangements around the TM 1b/TM 6a interface. PMID- 18381284 TI - Dopamine D1 receptor-induced signaling through TrkB receptors in striatal neurons. AB - In addition to its role as a neurotransmitter, dopamine can stimulate neurite outgrowth and morphological effects upon primary neurons. To investigate the signal transduction mechanisms used by dopamine in developing striatal neurons, we focused upon the effects of activating the dopamine D1 receptor. Using the D1 receptor agonist SKF38393, we found that Trk neurotrophin receptors were activated in embryonic day 18 striatal neurons. K-252a, a Trk tyrosine kinase inhibitor, and a dopamine D1 receptor antagonist could block the effects of SKF38393. The increase in TrkB phosphorylation was not the result of increased neurotrophin production. Induction of TrkB activity by SKF38393 was accompanied by the phosphorylation of several Trk signaling proteins, including phospholipase Cgamma, Akt, and MAPK. Biotinylation experiments followed by immunostaining by phospho-TrkB-specific antibodies indicated that the mechanism involved increased TrkB surface expression by dopamine D1 receptor activation. This increase in cell surface TrkB expression was dependent upon an increase in intracellular Ca(2+). These results indicate that stimulation of dopamine D1 receptors can be coupled to the neurotrophin receptor signaling to mediate the effects of dopamine upon striatal neurons. PMID- 18381287 TI - Chronic suppression of acetyl-CoA carboxylase 1 in beta-cells impairs insulin secretion via inhibition of glucose rather than lipid metabolism. AB - Acetyl-CoA carboxylase 1 (ACC1) currently is being investigated as a target for treatment of obesity-associated dyslipidemia and insulin resistance. To investigate the effects of ACC1 inhibition on insulin secretion, three small interfering RNA (siRNA) duplexes targeting ACC1 (siACC1) were transfected into the INS-1-derived cell line, 832/13; the most efficacious duplex was also cloned into an adenovirus and used to transduce isolated rat islets. Delivery of the siACC1 duplexes decreased ACC1 mRNA by 60-80% in 832/13 cells and islets and enzyme activity by 46% compared with cells treated with a non-targeted siRNA. Delivery of siACC1 decreased glucose-stimulated insulin secretion (GSIS) by 70% in 832/13 cells and by 33% in islets. Surprisingly, siACC1 treatment decreased glucose oxidation by 49%, and the ATP:ADP ratio by 52%, accompanied by clear decreases in pyruvate cycling activity and tricarboxylic acid cycle intermediates. Exposure of siACC1-treated cells to the pyruvate cycling substrate dimethylmalate restored GSIS to normal without recovery of the depressed ATP:ADP ratio. In siACC1-treated cells, glucokinase protein levels were decreased by 25%, which correlated with a 36% decrease in glycogen synthesis and a 33% decrease in glycolytic flux. Furthermore, acute addition of the ACC1 inhibitor 5 (tetradecyloxy)-2-furoic acid (TOFA) to beta-cells suppressed [(14)C]glucose incorporation into lipids but had no effect on GSIS, whereas chronic TOFA administration suppressed GSIS and glucose metabolism. In sum, chronic, but not acute, suppression of ACC1 activity impairs GSIS via inhibition of glucose rather than lipid metabolism. These findings raise concerns about the use of ACC inhibitors for diabetes therapy. PMID- 18381289 TI - Molecular basis of histone H3K4me3 recognition by ING4. AB - The inhibitors of growth (ING) family of tumor suppressors consists of five homologous proteins involved in chromatin remodeling. They form part of different acetylation and deacetylation complexes and are thought to direct them to specific regions of the chromatin, through the recognition of H3K4me3 (trimethylated K4 in the histone 3 tail) by their conserved plant homeodomain (PHD). We have determined the crystal structure of ING4-PHD bound to H3K4me3, which reveals a tight complex stabilized by numerous interactions. NMR shows that there is a reduction in the backbone mobility on the regions of the PHD that participate in the peptide binding, and binding affinities differ depending on histone tail lengths Thermodynamic analysis reveals that the discrimination in favor of methylated lysine is entropy-driven, contrary to what has been described for chromodomains. The molecular basis of H3K4me3 recognition by ING4 differs from that of ING2, which is consistent with their different affinities for methylated histone tails. These differences suggest a distinct role in transcriptional regulation for these two ING family members because of the antagonistic effect of the complexes that they recruit onto chromatin. Our results illustrate the versatility of PHD fingers as readers of the histone code. PMID- 18381288 TI - Alpha1 soluble guanylyl cyclase (sGC) splice forms as potential regulators of human sGC activity. AB - Soluble guanylyl cyclase (sGC), a key protein in the NO/cGMP signaling pathway, is an obligatory heterodimeric protein composed of one alpha- and one beta subunit. The alpha(1)/beta(1) sGC heterodimer is the predominant form expressed in various tissues and is regarded as the major isoform mediating NO-dependent effects such as vasodilation. We have identified three new alpha(1) sGC protein variants generated by alternative splicing. The 363 residue N1-alpha(1) sGC splice variant contains the regulatory domain, but lacks the catalytic domain. The shorter N2-alpha(1) sGC maintains 126 N-terminal residues and gains an additional 17 unique residues. The C-alpha(1) sGC variant lacks 240 N-terminal amino acids, but maintains a part of the regulatory domain and the entire catalytic domain. Q-PCR of N1-alpha(1), N2-alpha(1) sGC mRNA levels together with RT-PCR analysis for C-alpha(1) sGC demonstrated that the expression of the alpha(1) sGC splice forms vary in different human tissues indicative of tissue specific regulation. Functional analysis of the N1-alpha(1) sGC demonstrated that this protein has a dominant-negative effect on the activity of sGC when coexpressed with the alpha(1)/beta(1) heterodimer. The C-alpha(1) sGC variant heterodimerizes with the beta(1) subunit and produces a fully functional NO- and BAY41-2272-sensitive enzyme. We also found that despite identical susceptibility to inhibition by ODQ, intracellular levels of the 54-kDa C-alpha(1) band did not change in response to ODQ treatments, while the level of 83 kDa alpha(1) band was significantly affected by ODQ. These studies suggest that modulation of the level and diversity of splice forms may represent novel mechanisms modulating the function of sGC in different human tissues. PMID- 18381290 TI - The synthesis of UDP-N-acetylglucosamine is essential for bloodstream form trypanosoma brucei in vitro and in vivo and UDP-N-acetylglucosamine starvation reveals a hierarchy in parasite protein glycosylation. AB - A gene encoding Trypanosoma brucei UDP-N-acetylglucosamine pyrophosphorylase was identified, and the recombinant protein was shown to have enzymatic activity. The parasite enzyme is unusual in having a strict substrate specificity for N acetylglucosamine 1-phosphate and in being located inside a peroxisome-like microbody, the glycosome. A bloodstream form T. brucei conditional null mutant was constructed and shown to be unable to sustain growth in vitro or in vivo under nonpermissive conditions, demonstrating that there are no alternative metabolic or nutritional routes to UDP-N-acetylglucosamine and providing a genetic validation for the enzyme as a potential drug target. The conditional null mutant was also used to investigate the effects of N-acetylglucosamine starvation in the parasite. After 48 h under nonpermissive conditions, about 24 h before cell lysis, the status of parasite glycoprotein glycosylation was assessed. Under these conditions, UDP-N-acetylglucosamine levels were less than 5% of wild type. Lectin blotting and fluorescence microscopy with tomato lectin revealed that poly-N-acetyllactosamine structures were greatly reduced in the parasite. The principal parasite surface coat component, the variant surface glycoprotein, was also analyzed. Endoglycosidase digestions and mass spectrometry showed that, under UDP-N-acetylglucosamine starvation, the variant surface glycoprotein was specifically underglycosylated at its C-terminal Asn-428 N glycosylation site. The significance of this finding, with respect to the hierarchy of site-specific N-glycosylation in T. brucei, is discussed. PMID- 18381291 TI - Structural and functional consequences of tyrosine phosphorylation in the LRP1 cytoplasmic domain. AB - The cytoplasmic domain of LRP1 contains two NPXY motifs that have been shown to interact with signaling proteins. In previous work, we showed that Tyr(4507) in the distal NPXY motif is phosphorylated by v-Src, whereas denaturation of the protein was required for phosphorylation of Tyr(4473) in the membraneproximal NPXY motif. Amide H/D exchange studies reveal that the distal NPXY motif is fully solvent-exposed, whereas the proximal one is not. Phosphopeptide mapping combined with in vitro and in vivo kinase experiments show that Tyr(4473) can be phosphorylated, but only if Tyr(4507) is phosphorylated or substituted with glutamic acid. Amide H/D exchange experiments indicate that solvent accessibility increases across the entire LRP1 cytoplasmic region upon phosphorylation at Tyr(4507); in particular the NPXY(4473) motif becomes much more exposed. This differential phosphorylation is functionally relevant: binding of Snx17, which is known to bind at the proximal NPXY motif, is inhibited by phosphorylation at Tyr(4473). Conversely, Shp2 binds most strongly when both of the NPXY motifs in LRP1 are phosphorylated. PMID- 18381292 TI - RAM-induced allostery facilitates assembly of a notch pathway active transcription complex. AB - The Notch pathway is a conserved cell-to-cell signaling mechanism, in which extracellular signals are transduced into transcriptional outputs through the nuclear effector CSL. CSL is converted from a repressor to an activator through the formation of the CSL-NotchIC-Mastermind ternary complex. The RAM (RBP-J associated molecule) domain of NotchIC avidly interacts with CSL; however, its role in assembly of the CSL-NotchIC-Mastermind ternary complex is not understood. Here we provide a comprehensive thermodynamic, structural, and biochemical analysis of the RAM-CSL interaction for components from both mouse and worm. Our binding data show that RAM and CSL form a high affinity complex in the presence or absence of DNA. Our structural studies reveal a striking distal conformational change in CSL upon RAM binding, which creates a docking site for Mastermind to bind to the complex. Finally, we show that the addition of a RAM peptide in trans facilitates formation of the CSL-NotchIC-Mastermind ternary complex in vitro. PMID- 18381293 TI - Bioluminescence resonance energy transfer assays reveal ligand-specific conformational changes within preformed signaling complexes containing delta opioid receptors and heterotrimeric G proteins. AB - Heptahelical receptors communicate extracellular information to the cytosolic compartment by binding an extensive variety of ligands. They do so through conformational changes that propagate to intracellular signaling partners as the receptor switches from a resting to an active conformation. This active state has been classically considered unique and responsible for regulation of all signaling pathways controlled by a receptor. However, recent functional studies have challenged this notion and called for a paradigm where receptors would exist in more than one signaling conformation. This study used bioluminescence resonance energy transfer assays in combination with ligands of different functional profiles to provide in vivo physical evidence of conformational diversity of delta-opioid receptors (DORs). DORs and alpha(i1)beta(1)gamma(2) G protein subunits were tagged with Luc or green fluorescent protein to produce bioluminescence resonance energy transfer pairs that allowed monitoring DOR-G protein interactions from different vantage points. Results showed that DORs and heterotrimeric G proteins formed a constitutive complex that underwent structural reorganization upon ligand binding. Conformational rearrangements could not be explained by a two-state model, supporting the idea that DORs adopt ligand specific conformations. In addition, conformational diversity encoded by the receptor was conveyed to the interaction among heterotrimeric subunits. The existence of multiple active receptor states has implications for the way we conceive specificity of signal transduction. PMID- 18381295 TI - Actual incidences of road casualties, and their injury severity, modelled from police and hospital data, France. AB - BACKGROUND: Nation-wide road casualty figures usually come from police data. In France, as in many developed countries, the reporting of fatalities is almost complete but the reporting of non-fatal casualties is rather low. It is moreover strongly biased. Valid estimates are needed. METHODS: Using the capture-recapture method on police data and on a road trauma registry covering a large county of 1.6 million inhabitants, we estimate police under-reporting correction factors that account for unregistered casualties. These correction factors are then applied to the nation-wide police data, with standardization on under-reporting bias factors. RESULTS: In 2004, whereas the police report 108,727 non-fatally injured, the estimation yields 400,200. Over the 1996-2004 study period, the average annual estimated incidence is 871/100,000 for all injured (3.4 times the police incidence), 232/100,000 for hospitalized, 103/100,000 for seriously injured (2.2 times the police incidence) and 12.6/100,000 for casualties with long-term major impairment. The incidence of seriously injured (NISS 9+) is 11.3/100,000 for pedestrians, 9.5/100,000 for cyclists, 36.3/100,000 for motorized two-wheel users and 42.5/100,000 for car users. CONCLUSIONS: The estimated incidences are much higher than the police-based ones. This changes the scale of the road injuries issue. The risk of suffering a major impairment from a road crash is equal to the risk of being killed. Motorized two-wheel users experience a large burden of traffic casualties, much larger than that indicated by police data. The approach used can be reproduced in other countries, if an additional medical registration exists. PMID- 18381296 TI - Low HIV-testing rates and awareness of HIV infection among high-risk heterosexual STI clinic attendees in The Netherlands. AB - OBJECTIVES: Since 1999, HIV testing is routinely offered to all attendees of the sexually transmitted infections (STI) outpatient clinic in Amsterdam, the Netherlands. This study evaluates whether this more active HIV-testing policy increased uptake of HIV testing and awareness of an HIV-positive serostatus among heterosexual attendees. METHODS: In addition to routine data collected at each STI consultation, data from half-yearly HIV surveys were used from 1994 to 2004. During each survey period, 1000 consecutive attendees are enrolled voluntary and anonymously for HIV testing and are interviewed on previous HIV testing and outcome. Trends in and predictors for uptake of HIV testing as offered during routine STI consultation were analysed by logistic regression. Trends in awareness of an HIV-positive serostatus as obtained from the anonymous HIV surveys were likewise analysed. RESULTS: The percentage of heterosexual attendees opting for an HIV test during consultation increased from 13% in 1996 to 56% in 2004. However, the proportion of individuals aware of their HIV infection did not change over time and only a minority (19%) of the 108 attendees found HIV positive in the anonymous surveys were aware of their HIV infection. Persons being or visiting a commercial sex worker, having a non-Dutch ethnicity, lacking health insurance and having an STI diagnosed were less likely to opt for an HIV test. CONCLUSIONS: Although heterosexual attendees increased their uptake of HIV testing during STI consultation over time, uptake of testing by attendees at risk for HIV infection, such as those infected with an STI, remained low. As a result, the percentage of persons aware of their HIV infection remained low, posing a risk for their individual health and for ongoing HIV transmission. Current testing strategies, therefore, misses the group that most needs testing. Based on these results, 'opt-out' HIV testing is now the standard procedure at the Amsterdam STI clinic. PMID- 18381294 TI - Involvement of hypoxia-inducing factor-1alpha-dependent plasminogen activator inhibitor-1 up-regulation in Cyr61/CCN1-induced gastric cancer cell invasion. AB - Cysteine-rich 61 (Cyr61/CCN1), one of the members of CCN family, has been implicated in the progression of human malignancies. Previously, our studies have demonstrated that Cyr61/CCN1 has a role in promoting gastric cancer cell invasion, but the mechanism is not clear yet. Here, we found that hypoxia inducing factor-1alpha (HIF-1alpha) protein, but not mRNA, expression was significantly elevated in gastric cancer cells overexpressing Cyr61. Supportively, a profound reduction of endogenous HIF-1alpha protein was noted in one highly invasive cell line, TSGH, when transfected with antisense Cyr61. By comparison, the induction kinetics of HIF-1alpha protein by recombinant Cyr61 (rCyr61) was distinct from that of insulin-like growth factor-1 and CoCl(2) treatment, both well known for induction of HIF-1alpha. Using cycloheximide and MG132, we demonstrated that the Cyr61-mediated HIF-1alpha up-regulation was through de novo protein synthesis, rather than increased protein stability. rCyr61 could also activate the PI3K/AKT/mTOR and ERK1/2 signaling pathways, both of which were essential for HIF-1alpha protein accumulation. Blockage of HIF 1alpha activity in Cyr61-expressing cells by transfecting with a dominant negative (DN)-HIF-1alpha strongly inhibited their invasion ability, suggesting that elevation in HIF-1alpha protein is vital for Cyr61-mediated gastric cancer cell invasion. In addition, several HIF-1alpha-regulated invasiveness genes were examined, and we found that only plasminogen activator inhibitor-1 (PAI-1) showed a significant increase in mRNA and protein levels in cells overexpressing Cyr61. Treatment with PAI-1-specific antisense oligonucleotides or function-neutralizing antibodies abolished the invasion ability of the Cyr61-overexpressing cells. Transfection with dominant negative-HIF-1alpha to block HIF-1alpha activity also effectively reduced the elevated PAI-1 level. In conclusion, our data provide a detailed mechanism by which Cyr61 promoted gastric cancer cell invasive ability via an HIF-1alpha-dependent up-regulation of PAI-1. PMID- 18381297 TI - Still "not satisfied"...Yet. PMID- 18381298 TI - Invited commentary. PMID- 18381299 TI - On "intertester reliability and validity of motion assessments..." Landel et al. Phys Ther. 2008;88:43-49. PMID- 18381300 TI - On "journal publication productivity..." Richter et al. Phys Ther. 2008;88:376 386. PMID- 18381301 TI - On "journal publication productivity..." Richter et al. Phys Ther. 2008;88:376 386. PMID- 18381302 TI - On "journal publication productivity..." Richter et al. Phys Ther. 2008;88:376 386. PMID- 18381303 TI - Technology assessment and adoption in orthopaedics: lessons learned. PMID- 18381304 TI - Geographic variation in device use for intertrochanteric hip fractures. AB - BACKGROUND: Hip fractures in the elderly are a common and costly problem, with intertrochanteric fractures accounting for almost half of these fractures. Most intertrochanteric fractures are treated with either a plate-and-screw device or an intramedullary nail device. We assessed the degree of geographic variation in use of intramedullary nailing for intertrochanteric femoral fractures among Medicare beneficiaries between 2000 and 2002. METHODS: Medicare 100% files (hospital and physician claims, and enrollment) for 2000 through 2002 were used to identify beneficiaries, sixty-five years of age or older, who had undergone inpatient surgery for the treatment of an intertrochanteric femoral fracture with a plate-and-screw device or an intramedullary nail. We used multiple logistic regression analysis to model the use of an intramedullary nail (as opposed to a plate-and-screw device) by state and year, after adjusting for patient age, sex, race, subtrochanteric fracture, comorbidities, and Medicaid-administered assistance. The odds ratios of receiving an intramedullary nail device are reported. The adjusted state rates of intramedullary nailing per 100 Medicare patients with an intertrochanteric fracture are reported for 2000 through 2002. RESULTS: In this study, 212,821 claims for operations to treat patients with an intertrochanteric fracture from 2000 through 2002 met the inclusion criteria. There was considerable geographic variation in intramedullary nail use by state across all years. The mean adjusted intramedullary nailing rate per 100 Medicare patients with an intertrochanteric fracture increased nationally from 7.84 in 2000 to 16.98 in 2002. In 2000, surgeons in sixteen states used an intramedullary nail in fewer than one of every twenty Medicare patients with an intertrochanteric fracture. By 2002, surgeons in only two states used an intramedullary nail in fewer than one of every twenty patients with an intertrochanteric fracture, and in eight states they used an intramedullary nail in more than one of every four patients with an intertrochanteric fracture. CONCLUSIONS: There was substantial geographic variation in the use of intramedullary nailing by state from 2000 through 2002 that was largely not explained by patient-related factors. PMID- 18381305 TI - Nail or plate fixation of intertrochanteric hip fractures: changing pattern of practice. A review of the American Board of Orthopaedic Surgery Database. AB - BACKGROUND: A new method of fixation for intertrochanteric hip fractures that involves the use of an intramedullary nail that interlocks proximally into the femoral head was introduced in the early 1990s. Anecdotal observation of practice patterns during the Part II (oral) American Board of Orthopaedic Surgery examination suggested that the use of this method had increased substantially in recent years in comparison with the more traditional sliding compression screw technique. A study of the Part II database was undertaken to detect changing patterns of care for intertrochanteric fractures. METHODS: During the process of Board certification, candidates for the Part II (oral) examination submit a six month surgical case list and patient data into a secure database. The database was searched for all intertrochanteric fractures (International Classification of Diseases, Ninth Revision, code 820.20 or 820.21) over a seven-year period (1999 through 2006). The cases were categorized by intramedullary nail or plate fixation on the basis of surgeon-reported Current Procedural Terminology codes. Relative utilization of the two devices was analyzed according to year and region, and the devices were compared in terms of complications and outcomes. RESULTS: A dramatic change in practice was demonstrated, with the intramedullary nail fixation rate increasing from 3% in 1999 to 67% in 2006. Regional variation was substantial. The highest rate of utilization of intramedullary nail fixation was recorded by candidates from the South, Southeast, and Southwest, who converted to the new technology faster than those in the Northeast, Northwest, and Midwest. Overall, patients managed with plate fixation had slightly less pain and deformity in comparison with those managed with intramedullary nailing, with no significant differences being identified in terms of function or satisfaction. Patients managed with intramedullary nailing had more procedure-related complications, particularly bone fracture. CONCLUSIONS: From 1999 to 2006, a dramatic change in surgeon preference for the fixation device used for the treatment of intertrochanteric fractures has occurred among young orthopaedic surgeons. This change has occurred despite a lack of evidence in the literature supporting the change and in the face of the potential for more complications. PMID- 18381306 TI - Primary arthroscopic stabilization for a first-time anterior dislocation of the shoulder. A randomized, double-blind trial. AB - BACKGROUND: Anterior dislocation of the glenohumeral joint in younger patients is associated with a high risk of recurrence and persistent functional deficits. The aim of this study was to assess the efficacy of a primary arthroscopic Bankart repair, while controlling for the therapeutic effects produced by the arthroscopic intervention and joint lavage. METHODS: In a single-center, double blind clinical trial, eighty-eight adult patients under thirty-five years of age who had sustained a primary anterior glenohumeral dislocation were randomized to receive either an arthroscopic examination and joint lavage alone or together with an anatomic repair of the Bankart lesion. Assessment of the rate of recurrent instability, functional outcome (with use of three scores), range of movement, patient satisfaction, direct health-service costs, and treatment complications was completed for eighty-four of these patients (forty-two in each group) during the subsequent two years. RESULTS: In the two years after the primary dislocation, the risk of a further dislocation was reduced by 76% and the risk of all recurrent instability was reduced by 82% in the Bankart repair group compared with the group that had arthroscopy and lavage alone. The functional scores were also better (p < 0.05), the treatment costs were lower (p = 0.012), and patient satisfaction was higher (p < 0.001) after arthroscopic repair. The improved functional outcome appeared to be mediated through the prevention of instability since the functional outcome in patients with stable shoulders was similar, irrespective of the initial treatment allocation. The patients who had a Bankart repair and played contact sports were also more likely to have returned to their sport at two years (relative risk = 3.4, p = 0.007). CONCLUSIONS: Following a first-time anterior dislocation of the shoulder, there is a marked treatment benefit from primary arthroscopic repair of a Bankart lesion, which is distinct from the so-called background therapeutic effect of the arthroscopic examination and lavage of the joint. However, primary repair does not appear to confer a functional benefit to patients with a stable shoulder at two years after the dislocation. PMID- 18381307 TI - A randomized clinical trial comparing intralesional bone marrow and steroid injections for simple bone cysts. AB - BACKGROUND: Simple bone cysts are common benign lesions in growing children that predispose them to fracture and are sometimes painful. The purpose of this trial was to compare rates of healing of simple bone cysts treated with intralesional injections of bone marrow with rates of healing of those treated with methylprednisolone acetate. METHODS: Of ninety patients randomly allocated to treatment with either a bone-marrow or a methylprednisolone acetate injection, seventy-seven were followed for two years. The primary outcome, determined by a radiologist who was blind to the type of treatment, was radiographic evidence of healing. The cyst was judged to be either not healed (grade 1 [a clearly visible cyst] or grade 2 [a cyst that was visible but multilocular and opaque]) or healed (grade 3 [sclerosis around or within a partially visible cyst] or grade 4 [complete healing with obliteration of the cyst]). Patient function was assessed with use of the Activity Scale for Kids, and pain was assessed with the Oucher Scale. RESULTS: Sixteen (42%) of the thirty-eight cysts treated with methylprednisolone acetate healed, and nine (23%) of the thirty-nine cysts treated with bone marrow healed (p = 0.01). There was no significant difference between the treatment groups (p > 0.09) with respect to function, pain, number of injections, additional fractures, or complications. CONCLUSIONS: Although the rate of healing of simple bone cysts was low following injection of either bone marrow or methylprednisolone, the latter provided superior healing rates. PMID- 18381308 TI - Intra-articular block compared with conscious sedation for closed reduction of ankle fracture-dislocations. A prospective randomized trial. AB - BACKGROUND: Ankle fracture-dislocations require urgent reduction to protect the soft tissues, to minimize articular injury, and to allow swelling to decrease. Conscious sedation is commonly used to provide analgesia for closed reduction of this injury. We hypothesized that an intra-articular block of the ankle would provide similar analgesia and the ability to reduce the ankle with a lower risk than conscious sedation. METHODS: Between September 2005 and January 2007, forty two patients with an ankle fracture-dislocation presented to our emergency department and were enrolled in a prospective randomized study. The patients were given either conscious sedation or an intra-articular lidocaine block for the reduction and for the application of a plaster splint. After the reduction maneuver, the patients used a visual analog pain scale to rate the level of pain before, during, and after the procedure, from 1 (no pain) to 10 (severe pain). The senior authors reviewed the injury and reduction radiographs to confirm the reduction of the ankle joint. RESULTS: Twenty-one patients were randomized to each group. There was no difference in demographic data or fracture patterns between the groups. Both the sedation and the block reduced the pain to a similar degree. The pain reduction (the initial pain level minus the level of pain after medication was given or injected) was an average (and standard deviation) of 4.6 +/- 3.3 for the block group and 4.2 +/- 3.5 for the sedation group (p = 0.64). The average change in the level of pain between the initial presentation and during the reduction was 3.6 +/- 3.8 for the block group and 4.1 +/- 3.3 for the sedation group. Overall, there was no difference in analgesia provided by these two methods (p = 0.71). An acceptable reduction was achieved for forty-one of the forty-two patients with one failure in the sedation group. The average time for ankle reduction and stabilization in a splint was 81.5 minutes for the sedation group and 63.8 minutes for the block group. CONCLUSIONS: Compared with conscious sedation, an intra-articular lidocaine block provides a similar degree of analgesia and sufficient analgesia to achieve closed reduction of ankle fracture dislocations. PMID- 18381309 TI - Anterior cruciate ligament reconstruction in patients who have excessive joint laxity. AB - BACKGROUND: The purpose of the present study was to compare the clinical results of anterior cruciate ligament reconstruction with use of different grafts in patients with or without excessive joint laxity. METHODS: We retrospectively studied eighty-three patients (forty-eight men and thirty-five women) who had undergone anterior cruciate ligament reconstruction between June 2002 and August 2004. Of the thirty-one patients who had generalized joint laxity, twenty were managed with an autologous bone-patellar tendon-bone graft and eleven were managed with a four-bundle hamstring (semitendinosus-gracilis tendon) graft. Of the fifty-two patients who had normal joint laxity, thirty-three were managed with a bone-patellar tendon-bone graft and nineteen were managed with a hamstring graft. Clinical results were evaluated by means of an examination at a minimum of twenty-four months after surgery. RESULTS: Among the patients who had normal joint laxity, the mean side-to-side difference in anterior tibial translation on testing with the KT-2000 arthrometer at a minimum of twenty-four months postoperatively was 2.7 +/- 1.9 mm in the bone-patellar tendon-bone graft group and 3.5 +/- 1.7 mm in the hamstring graft group. This difference was not significant. The mean Lysholm score was 91 +/- 7 points in the bone-patellar tendon-bone graft group and 85 +/- 10 points in the hamstring graft group (p = 0.492). The side-to-side difference in anterior translation had an inverse correlation with the Lysholm score (beta = -0.604, p < 0.001). Among the patients who had excessive joint laxity, the mean side-to-side difference in anterior tibial translation was 3.4 +/- 1.5 mm in the bone-patellar tendon-bone graft group and 4.5 +/- 2.0 mm in the hamstring graft group (p = 0.036). The mean Lysholm score was 89 +/- 7 points in the bone-patellar tendon-bone group and 79 +/- 12 points in the hamstring group (p = 0.015). The side-to-side difference in anterior translation had an inverse correlation with the Lysholm score (beta = 0.708, p < 0.001). CONCLUSIONS: In patients who have excessive joint laxity, the two-year outcomes of anterior cruciate ligament reconstruction with bone-patellar tendon-bone grafts are better than those with four-bundle hamstring grafts in terms of both side-to-side anterior laxity and clinical results. PMID- 18381310 TI - Weight loss in overweight and obese patients following successful lumbar decompression. AB - BACKGROUND: Neurogenic claudication secondary to lumbar stenosis is often cited by overweight and obese patients as a factor limiting their ability to lose weight. Many patients believe that they will be able to increase their activity and subsequently lose weight following relief of symptoms. The objective of this study was to evaluate weight loss in overweight and obese patients who obtained substantial pain relief after lumbar decompression surgery for spinal stenosis. METHODS: Changes in the body weight and body mass index of overweight and obese patients after lumbar decompression surgery were assessed at a mean of 34.4 months postoperatively. Sixty-three patients (thirty-seven men and twenty-six women with a mean age of 53.4 years) were included in the study. Preoperative and postoperative body weight and body mass indices were calculated, and Zurich Claudication Questionnaire (ZCQ) Symptom Severity and Physical Function scores were obtained. RESULTS: The ZCQ Symptom Severity and Physical Function scores significantly improved, by a mean of 56.4% and 53.0%, respectively. At the time of follow-up, both the mean body weight and the mean body mass index significantly increased, by 2.48 kg and 0.83 kg/m(2), respectively. Overall, 35% of the patients gained >or=5% of their preoperative body weight, 6% of the patients lost >or=5% of their preoperative body weight, and 59% remained within 5% of their preoperative body weight. CONCLUSIONS: The majority of overweight and obese patients maintain or increase their body weight and body mass index following successful lumbar decompression surgery. Substantial relief of symptoms and functional improvements do not appear to help overweight or obese patients to lose weight. This suggests that obesity is an independent disease and not simply a function of symptomatic spinal stenosis, and patients should be counseled regarding these expectations. PMID- 18381311 TI - Characteristics of ossified lesions in the upper cervical spine associated with ossification of the posterior longitudinal ligament in the lower cervical spine. AB - BACKGROUND: Ossified lesions of the upper cervical spine behind the dens may cause cervical myelopathy. We investigated the prevalence of ossified lesions in patients with ossification of the posterior longitudinal ligament in order to clarify the characteristics of these patients. METHODS: Fifty-six consecutive patients with ossification of the posterior longitudinal ligament in the cervical spine were included in the study. Multidetector computed tomography images of the whole spine were made. Ossified lesions of the upper cervical spine were defined as ossified lesions behind the dens as seen on axial computed tomography images. The prevalence of ossified lesions at the first cervical level was analyzed. The clinical and radiographic parameters were compared between the patients with ossified lesions at the first cervical level and the patients without such lesions. RESULTS: Fourteen patients (25%) had ossified lesions of the upper cervical spine. The continuous type of ossification of the posterior longitudinal ligament in the lower cervical spine was common and the segmental type was rare in this group as compared with the types of ossification in the group without ossified lesions at the first cervical level. The patients with first cervical level ossification more frequently had ossification of the posterior longitudinal ligament in the thoracic and/or lumbar spine, compared with the patients without upper cervical ossification. One patient had cervical myelopathy as a result of the upper cervical ossified lesion. CONCLUSIONS: Computed tomography imaging demonstrated the presence of ossified lesions behind the dens in 25% of patients with subaxial ossification of the posterior longitudinal ligament. These upper cervical ossified lesions were commonly seen in patients with multiple-level ossification of the posterior longitudinal ligament. When upper cervical ossified lesions cause severe narrowing of the spinal canal, they should be considered a potential cause of cervical myelopathy. PMID- 18381312 TI - Charcot arthropathy and immobilization in a weight-bearing total contact cast. AB - BACKGROUND: The standard management for Eichenholtz Stage-I Charcot arthropathy has been with non-weight-bearing total contact casts. The purpose of this study was to evaluate the results of the use of weight-bearing total contact casts for similar patients. METHODS: Twenty-seven patients with Charcot arthropathy of the foot and ankle were studied prospectively over a period of eighteen years, from 1988 to 2006. The average duration of follow-up was 5.5 years. Of the twenty seven patients, twenty-six had diabetes mellitus. Total contact casts were used to treat thirty-four feet with Eichenholtz Stage-I or early Stage-II Charcot arthropathy. These patients were allowed to bear weight as tolerated. Casts were changed at weekly intervals and were worn until resolution of the acute stage of the disease. RESULTS: No deleterious effect from weight-bearing, specifically with regard to skin ulceration or rapid deterioration of the osseous architecture, was observed in thirty-three of the thirty-four feet. CONCLUSIONS: Immobilization in a weight-bearing total contact cast appears to be a safe method of treatment of acute Eichenholtz Stage-I Charcot arthropathy of the foot and ankle. PMID- 18381313 TI - Autograft contamination during preparation for anterior cruciate ligament reconstruction. AB - BACKGROUND: The autograft preparation process for anterior cruciate ligament reconstruction has a potential for graft contamination. The purpose of this study was to evaluate the possibility of contamination of the bone-patellar tendon-bone and hamstring tendon autograft during preparation for anterior cruciate ligament reconstruction. METHODS: A primary isolated reconstruction of the anterior cruciate ligament with use of bone-patellar tendon-bone autograft (thirty patients) and hamstring tendon autograft (thirty patients) was performed in a prospective, consecutive series of patients. Three tissue samples were obtained for culture from each graft at different time-intervals during the graft preparation. In addition, the erythrocyte sedimentation rate and the C-reactive protein level were evaluated preoperatively and on the third, seventh, and twentieth postoperative days, and the clinical course of all patients was monitored. RESULTS: The time needed for graft preparation was significantly longer for hamstring autografts (nineteen minutes) than for bone-patellar tendon bone autografts (ten minutes) (p = 0.032). In the hamstring group, cultures of graft tissue from four patients (13%) were positive for bacteria. In the bone patellar tendon-bone group, cultures of graft tissue from three patients (10%) were positive for bacteria; the difference between groups was not significant (p = 0.923). No patient had development of a postoperative infection. There were no differences between patients with a contaminated graft and those with an uncontaminated graft with regard to postoperative changes in the erythrocyte sedimentation rate or the C-reactive protein level at all time-intervals. CONCLUSIONS: A high rate (12%) of autograft contamination can be expected during autograft preparation for anterior cruciate ligament reconstruction. The contamination rate is almost equal for both bone-patellar tendon-bone and hamstring tendon autografts. We could not identify an association between contaminated grafts implanted in the knee and postoperative inflammatory markers such as the erythrocyte sedimentation rate and the C-reactive protein level. PMID- 18381314 TI - Alignment deviation between bone resection and final implant positioning in computer-navigated total knee arthroplasty. AB - BACKGROUND: Computer-navigated total knee arthroplasty is aimed at improving accuracy in the positioning of prosthetic components and realigning the lower limb. The optimal position and orientation of the bone resection planes are targeted by the navigation system, but, after these are obtained, additional manual surgical actions, which may considerably affect final component alignment, are necessary for implantation. The aim of this study was to measure the alignment deviation caused by standard impaction of the tibial and femoral components following bone resections with use of navigation control. METHODS: Ninety-one primary total knee arthroplasties were performed with an image-free knee navigation system. The alignment of the tibial and femoral bone resections was measured in three planes during surgery by the instrumented probe of the system. The alignment measure was repeated after final tibial and femoral component implantation with cement. The alignment deviations between the two measures were considered the positioning error associated with the final manual implantation of the components. RESULTS: The alignment deviations between the bone resections and the subsequent implant placement were >1 degrees in the frontal plane of the femur and in the frontal and sagittal planes of the tibia in 20%, 11%, and 33% of the patients, respectively. The deviations were >2 degrees in 4%, 3%, and 9% of the patients, respectively. Deviations as large as 3 degrees were found at the tibia in the sagittal plane (the posterior slope). CONCLUSIONS: Positioning of the femoral and tibial components in total knee arthroplasty, which mainly involves cementation and impaction of the final components, can introduce a considerable error in alignment, regardless of how accurately the resection planes are made. After computer-navigated total knee arthroplasty, it would be useful therefore to check the alignment of the prosthetic component carefully before the cement hardens. PMID- 18381315 TI - A study of metatarsal fractures in children. AB - BACKGROUND: Metatarsal fractures are common in children. The aim of the present study was to analyze a consecutive series of metatarsal fractures in children and to describe the epidemiology of the fractures, the location of the fractures, and the mechanism of injury. METHODS: A consecutive series of 125 patients with metatarsal fractures who presented to one pediatric surgery department over a twenty-two-month period was evaluated with use of a questionnaire to define the mechanism of injury. All patients were followed until the fracture was healed. The specific location of the fractures and the mechanism of injury were identified. RESULTS: One hundred and twenty-five children (seventy-five boys and fifty girls; average age, 8.6 years [range, one to seventeen years]) presented with 166 metatarsal fractures. Major differences were found between children who were five years of age or less and those who were more than five years of age. In patients who were five years of age or less, the predominant location of the accident that had caused the fracture was inside the house and the primary mechanism was a fall from a height. In patients who were more than five years of age, most accidents occurred at sports facilities and were caused by a fall on a level surface. The most frequently fractured metatarsal in younger children was the first, whereas the most frequently fractured metatarsal in older children was the fifth. CONCLUSIONS: The mechanism of injury and clinical presentation of metatarsal fractures in children are age-dependent, with patients five years of age or less and those more than five years of age having different mechanisms of injury and different types of metatarsal fractures. PMID- 18381316 TI - The effect of postoperative range of motion on functional activities after posterior cruciate-retaining total knee arthroplasty. AB - BACKGROUND: Range of motion is recognized as an important indicator of the success of a total knee replacement; however, an optimal range of motion has yet to be defined. This study was designed to determine the optimal range of motion for knee function after total knee arthroplasty with a posterior cruciate retaining prosthesis. METHODS: We retrospectively reviewed 5556 primary total knee arthroplasties performed with posterior cruciate-retaining prostheses between 1983 and 2003. The relationship between postoperative range of motion and pain, walking ability, stair-climbing ability, and knee function scores was examined at three to five years postoperatively. The relationship between a postoperative flexion contracture or hyperextension and knee function was also examined. RESULTS: Patients with 128 degrees to 132 degrees of motion obtained the highest scores for pain, walking, and knee function and the highest Knee Society scores. The outcomes became substantially compromised with motion of <118 degrees . Patients with 133 degrees to 150 degrees of motion had the highest scores for stair-climbing. A postoperative flexion contracture and hyperextension were associated with lower scores for pain, walking, stair-climbing, and knee function. CONCLUSIONS: The best functional results following total knee arthroplasty are achieved with 128 degrees to 132 degrees of motion. A postoperative flexion contracture and hyperextension of >or=10 degrees are associated with a poorer outcome except that stair-climbing is improved with more motion. PMID- 18381317 TI - Replacement of the hip in children with a tumor in the proximal part of the femur. AB - BACKGROUND: Children rarely require hip replacement, and therefore very little is known about the most effective procedure for children who require hip replacement after proximal femoral resection for a tumor. METHODS: We reviewed the outcome of forty hip replacements in children between two and fifteen years old who had a proximal femoral replacement for malignant disease. Twenty-one children survived the illness and were followed for three to twenty-one years. Cemented acetabular components were used initially to restore hip function, but, in more recent procedures, unipolar replacements and uncemented implants were used. RESULTS: Children who were over the age of eleven years at the time of surgery had a rate of failure (defined as revision of the acetabular component) of 25% at ten years compared with a rate of 75% in younger children. Unipolar replacements in children of all ages failed by ten years, either because of pain or subluxation that led to revision. In children over the age of eleven years, both cemented and uncemented acetabular implants performed well. CONCLUSIONS: The preferred method for restoration of hip function in children under the age of eleven years following proximal femoral resection for a tumor remains unclear. We recommend that at the initial hip surgery an attempt should be made to cause as little damage as possible to the acetabulum, but most children will inevitably need revision surgery as they get older. We hope that this study will guide others in their decision-making with regard to this relatively rare condition. PMID- 18381318 TI - Noninvasive quantification of human nucleus pulposus pressure with use of T1rho weighted magnetic resonance imaging. AB - BACKGROUND: Early diagnosis is a challenge in the treatment of degenerative disc disease. A noninvasive biomarker detecting functional mechanics of the disc is needed. T1rho-weighted imaging, a spin-lock magnetic resonance imaging technique, has shown promise for meeting this need in in vivo studies demonstrating the clinical feasibility of evaluating both intervertebral discs and articular cartilage. The objectives of the present study were (1) to quantitatively determine the relationship between T1rho relaxation time and measures of nucleus pulposus mechanics, and (2) to evaluate whether the quantitative relationship of T1rho relaxation time with the degenerative grade and glycosaminoglycan content extend to more severe degeneration. It was hypothesized that the isometric swelling pressure and compressive modulus would be directly correlated with the T1rho relaxation time and the apparent permeability would be inversely correlated with the T1rho relaxation time. METHODS: Eight cadaver human lumbar spines were imaged to measure T1rho relaxation times. The nucleus pulposus tissue from the L1 disc through the S1 disc was tested in confined compression to determine the swelling pressure, compressive modulus, and permeability. The glycosaminoglycan and water contents were measured in adjacent tissue. Linear regression analyses were performed to examine the correlation between the T1rho relaxation time and the other measured variables. Mechanical properties and biochemical content were evaluated for differences associated with degeneration. RESULTS: A positive linear correlation was observed between the T1rho relaxation time on the images of the nucleus pulposus and the swelling pressure (r = 0.59), glycosaminoglycan content per dry weight (r = 0.69), glycosaminoglycan per wet weight (r = 0.49), and water content (r = 0.53). No significant correlations were observed between the T1rho relaxation time and the modulus or permeability. Similarly, the T1rho relaxation time, swelling pressure, glycosaminoglycan content per dry weight, and water content were significantly altered with degeneration, whereas the modulus and permeability were not. CONCLUSIONS: T1rho-weighted magnetic resonance imaging has a strong potential as a quantitative biomarker of the mechanical function of the nucleus pulposus and of disc degeneration. PMID- 18381319 TI - Lubricin distribution in the goat infraspinatus tendon: a basis for interfascicular lubrication. AB - BACKGROUND: This study was motivated by the need to better understand the tribology of the rotator cuff, as it could provide insights into degenerative processes and suggest therapeutic approaches to cuff disorders. The objective was to evaluate the distribution of a known lubricating protein in the infraspinatus tendon of the rotator cuff in adult goats. The hypothesis was that lubricin, also known as superficial zone protein or proteoglycan 4, serves as an interfascicular lubricant. METHODS: Eight infraspinatus tendons were resected from eight Spanish goats, and five patellar tendons and articular cartilage samples were also resected from five of the goats. Samples were processed for immunolocalization of lubricin with use of a purified monoclonal antibody and for histological analysis with Masson trichrome staining for collagen. The locations of lubricin within the tendon were documented, and measurements were made of the distance to which lubricin was detected in the tendon, relative to the humeral insertion site, and the depth of lubricin staining into the fibrocartilage of the fascicle bordering the humeral joint space. Images from polarized light microscopy were used to measure the fascicle diameter and the crimp length. RESULTS: Lubricin was prominent in layers separating the fascicles in the infraspinatus tendon, with occasional intrafascicular staining. The fibrocartilaginous portion of the fascicle bordering the humeral joint space displayed diffuse lubricin staining. The Masson trichrome staining of the collagen in this lubricin-containing fibrocartilage indicated that it was not being tensioned at the time of fixation, whereas the collagen contained in the body of the tendon was under tension. The crimp of the fascicles near the humeral joint side of the tendon displayed a shorter peak-to-peak length than the crimp of the fascicles in the superior region of the tendon. Only the surface of two of the five patellar tendon samples stained for lubricin; there was no staining within these ligaments. CONCLUSIONS: The sheaths of the fascicles of the infraspinatus tendon near the bone insertion site contain lubricin, indicating that this lubricating protein may be facilitating interfascicular movement. The fact that the crimp pattern of fascicles changes with location in the tendon provides support for the supposition that fascicles move relative to one another as the tendon is loaded, underscoring the importance of a lubricating protein in the layer separating the fascicles. PMID- 18381320 TI - Effect of varying hamstring tension on anterior cruciate ligament strain during in vitro impulsive knee flexion and compression loading. AB - BACKGROUND: The hamstring muscles are well positioned to limit both anterior tibial translation and anterior cruciate ligament strain during the knee flexion phase of a jump landing. We hypothesized that systematically increasing or decreasing hamstring tension during the knee flexion phase of a simulated jump landing would significantly affect peak relative strain in the anterior cruciate ligament. METHODS: Ten cadaveric knees from four male and six female donors (mean age [and standard deviation] at the time of death, 60.3 +/- 23.6 years) were mounted in a custom fixture to initially position the specimen in 25 degrees of knee flexion and simulate axial impulsive loading averaging 1700 N to cause an increase in knee flexion. Quadriceps, hamstring, and gastrocnemius muscle forces were simulated with use of pretensioned linear springs, with the tension in the hamstrings arranged to be increased, held constant, decreased, at "baseline," or absent during knee flexion. Impulsive loading applied along the tibia and femur was monitored with use of triaxial load transducers, while uniaxial load cells monitored quadriceps and medial and lateral hamstring forces. Relative strain in the anterior cruciate ligament was measured with use of a differential variable reluctance transducer, and tibiofemoral kinematics were measured optoelectronically. For each specimen, anterior cruciate ligament strains were recorded over eighty impact trials: ten preconditioning trials, ten "baseline" trials involving decreasing hamstring tension performed before and after three sets of ten trials conducted with increasing hamstring tension, constant hamstring tension, or no hamstring tension. Peak relative strains in the anterior cruciate ligament were normalized for comparison across specimens. RESULTS: Increasing hamstring force during the knee flexion landing phase decreased the peak relative strain in the anterior cruciate ligament by >70% compared with the baseline condition (p = 0.005). Neither a constant hamstring muscle force nor the absence of a hamstring force significantly changed the peak strain in the anterior cruciate ligament relative to the baseline condition. CONCLUSIONS: Increasing hamstring muscle force during the knee flexion phase of a simulated jump landing significantly reduces the peak relative strain in the anterior cruciate ligament in vitro. PMID- 18381321 TI - Effect of preoperative combined treatment with alendronate and calcitriol on fixation of hydroxyapatite-coated implants in ovariectomized rats. AB - BACKGROUND: Osteoporosis is a potential risk factor affecting implant stability following total joint arthroplasty. Bisphosphonates are potent inhibitors of osteoblastic bone resorption, and vitamin D is an important hormone involved in the regulation of calcium metabolism. We investigated the benefit of preoperative treatment with alendronate, vitamin D (calcitriol), or a combination of these substances on fixation of hydroxyapatite-coated implants in an ovariectomized rat model of osteoporosis. METHODS: Of sixty-four female rats in the study, fifty-six underwent ovariectomy and eight, a sham operation. A hydroxyapatite-coated femoral implant and an uncoated implant were implanted in all rats seven months after the ovariectomy or the sham operation. Of the fifty-six rats that underwent ovariectomy, eight served as the control group. The remaining forty-eight were treated with alendronate, calcitriol, or a combination of these agents either starting eight weeks before implantation of the rod or starting immediately after implantation. The treatment was continued until four weeks after the implantation. Four weeks after the implantation, the total and cancellous bone mineral density at the proximal part of the tibia and the shear strength of the bone-implant interface were measured in all rats. RESULTS: Although total bone mineral density increased in all of the alendronate-treated groups, compared with that in the ovariectomized control group, cancellous bone mineral density increased only in the group pretreated with both alendronate and calcitriol. The implant stability in the ovariectomized control rats was significantly decreased compared with that in the sham-operation rats. While treatment with alendronate only or calcitriol only did not improve the stability of the implants, treatment with a combination of alendronate and calcitriol, starting preoperatively, significantly improved the stability of the hydroxyapatite-coated implants. CONCLUSIONS: Treatment with a combination of alendronate and calcitriol, starting preoperatively, can improve cancellous bone mineral density and the stability of hydroxyapatite-coated implants in an osteoporotic rat. PMID- 18381322 TI - The effect of electrical fields on gene and protein expression in human osteoarthritic cartilage explants. AB - BACKGROUND: The destruction of cartilage in patients with osteoarthritis is a consequence of an imbalance between matrix synthesis and degradation. The purpose of the present study was to determine the effects of electrical stimulation on these processes in full-thickness osteoarthritic adult human articular cartilage explants. METHODS: Full-thickness articular cartilage explants from osteoarthritic adult human knee joints were cultured in the absence or presence of interleukin-1beta (IL-1beta) and in the absence or presence of a specifically defined capacitively coupled electrical signal for seven or fourteen days. Total collagen and proteoglycan production were assessed by means of hydroxyproline and hexosamine analyses, respectively. Quantitative real-time polymerase chain reaction assays were used to measure mRNA expression levels of aggrecan, type-II collagen, collagenase-1 (MMP-1), collagenase-3 (MMP-13), stromelysin-1 (MMP-3), aggrecanase-1 (ADAM-TS4), and aggrecanase-2 (ADAM-TS5). RESULTS: Electrical stimulation of cultured explants for seven or fourteen days resulted in significant increases (p < 0.007) in proteoglycan and collagen production and a highly significant upregulation (p 20 MeV) of these meters to several hundred million electron volts. High-energy quasi-monoenergetic beams utilised as part of this study were produced by the cyclotron facilities at the Universite Catholique de Louvain (33 and 60 MeV) and the T. Svedberg Laboratory ( 46, 95, 143 and 173 MeV). In addition, measurements were also conducted at the Los Alamos Neutron Science Center, 800 MeV spallation neutron source, in broad energy fields with an average energy of 345 MeV. For the sake of completeness, data collected between 2.5 and 19 MeV in monoenergetic neutron fields at the German Physikalisch-Technische Bundesanstalt (PTB) facility are also included in this study. PMID- 18381336 TI - Concentrations of radon and its daughter products in and around Bangalore city. AB - Indoor radon and its progeny levels were measured during 2005-06 in Bangalore rural district and in Bangalore City by using Solid State Nuclear Track Detector (SSNTD)-based twin cup dosemeters, and the activity of radium present in soils and rocks was measured by using HPGe detector. Fifty dwellings of different types were chosen for the measurement. The dosimeters containing the detector (LR-115 Type II Film) used in each house were fixed 2 m above the floor. After an exposure time of 90 days, films were etched to reveal tracks. From the track density, the concentrations of radon were evaluated. The value of radon concentration in the indoor air near granite quarries varies from 55 to 300 Bq.m( 3) with a median of 155 Bq.m(-3) and its progeny varies from 0.24 to 19.6 mWL with a median of 8.4 mWL. In Bangalore City, the concentration of radon varies from 18.4 to 110 Bq.m(-3) with a median of 45 Bq.m(-3) and its progeny varies from 1.62 to 11.24 mWL with a median of 4.15 mWL. Higher concentrations of radon and its progeny were observed in granite quarries compared with Bangalore City. The main reason for the higher indoor radon and its progeny concentration is due to the mining activity and the types of the bedrock. The concentration of radon mainly depends on the activity of radium present in soils and rocks and the types of building materials used. The activity of radium varies in granitic regions of Bangalore rural district from 42.0 to 163.6 Bq.kg(-1) with a median of 112.8 Bq.kg(-1). The concentrations of indoor radon and its daughter products and equivalent effective dose are discussed. PMID- 18381337 TI - Ethical issues, justification, referral criteria for budget limited and high-dose procedures. AB - This paper reviews some of the issues connected with questions of ethics, health economics, radiation dose and referral criteria arising from a workshop held under the auspices of the Sentinel Research Program FP6-012909. An extensive bibliography of further reading is included. PMID- 18381338 TI - QA/acceptance testing of DEXA X-ray systems used in bone mineral densitometry. AB - New developments in dual energy X-ray absorptiometry (DEXA) imaging technology [fan beam and cone beam (CB)] result in higher exposure levels, shorter scan times, increased patient throughput and increased shielding requirements. This study presents the results of a European survey detailing the number and location of DEXA systems in SENTINEL partner states and the QA (quality assurance) currently performed by physicists and operators in these centres. The results of a DEXA equipment survey based on an in-house developed QA protocol are presented. Measurements show that the total effective dose to the patient from a spine and dual femur DEXA examination on the latest generation DEXA systems is comparable with a few microSv at most. Scatter measurements showed that the use of a mobile lead screen for staff protection was necessary for fan and CB systems. Scattered dose from newer generation systems may also exceed the exposure limits for the general public so structural shielding may also be required. Considerable variation in the magnitude and annual repeatability of half value layer was noted between different models of DEXA scanners. A comparative study of BMD (bone mineral density) accuracy using the European Spine Phantom highlighted a deviation of up to 7% in BMD values between scanners of different manufacturers. PMID- 18381339 TI - Between channels and tears: aim at ROS to save the membrane of dystrophic fibres. PMID- 18381340 TI - Kv7 (KCNQ) potassium channels that are mutated in human diseases. PMID- 18381341 TI - A follow-up study of children hospitalised with community-acquired pneumonia. AB - OBJECTIVE: To investigate the outcome for children hospitalised with radiologically confirmed community-acquired pneumonia (CAP) DESIGN: Controlled follow-up study. SETTING: Community based in Newcastle upon Tyne, North Tyneside and Northumberland schools. PATIENTS: 103 cases of radiologically confirmed CAP a median of 5.6 years (range 4.4-7.4) after admission to Newcastle General Hospital, matched for sex and school class to a mean of two controls (n = 248). INTERVENTIONS: A respiratory questionnaire, clinical examination and spirometry measurements. MAIN OUTCOME MEASURES: Multiple regression was used to describe associations between explanatory variables, including CAP, and outcome variables: forced expiratory volume in 1 s percent predicted (FEV(1) %), forced vital capacity percent predicted (FVC %), persistent cough, doctor diagnosis of asthma and abnormal chest shape. RESULTS: Cases were 2.9 times more likely (95% CI 1.45 to 5.71, p = 0.020) than controls to have persistent cough and 5.5 times more likely to have an abnormal chest shape (95% CI 1.65 to 18.28, p = 0.005). Cases of an atopic parent had a 7.0% deficit in FEV(1) % predicted (95% CI -10.5 to 3.2, p<0.001) and a 4.4% deficit in FVC % predicted (95% CI -8.0 to -0.78, p = 0.017), but were not at increased risk of subsequent asthma. Cases of a non atopic parent were at increased risk of subsequent asthma (OR 4.8, 95% CI 1.43 to 16.34, p = 0.011) but not of deficit in lung function. CONCLUSIONS: CAP requiring admission to hospital is associated with deficits in lung function and persistent respiratory symptoms. This has implications for follow-up for which recommendations are currently lacking. Parental atopy may be a determinant of outcome. PMID- 18381342 TI - Adiposity measures and blood pressure in Chinese children and adolescents. AB - OBJECTIVE: To investigate the association of adiposity measures with blood pressure (BP) in Chinese children and adolescents. DESIGN: A cross-sectional study. PARTICIPANTS: 1330 boys and 1170 girls aged 6-18 years from a rural population-based cohort of twins studied in Anhui, China, 1998-2000. OUTCOME MEASURES: Adiposity measures included body mass index (BMI), total body fat and trunk fat assessed by dual-energy x-ray absorptiometry. BMI was divided into fat mass index (FMI) and lean mass index (LMI) in the analysis. Major outcomes included: systolic (S) and diastolic (D) BP. Both linear and logistic regressions were performed to assess gender-specific associations between various adiposity measures and BP, with adjustment for age and height. Generalised estimating equations were used to account for intra-twin pair correlations. RESULTS: Mean BMI and percentage body fat in children aged 6-11 years were 14.9 kg/m(2) and 9.7%, respectively; corresponding measures in children aged 12-18 years were 17.8 kg/m(2) and 14.2%. Adiposity measures were more strongly associated with SBP (p<0.05 in all age strata) than DBP (p<0.05 only in children aged 6-11 years). Both FMI (beta = 1.26-2.37) and LMI (beta = 1.00-1.71) are associated with SBP across age and gender strata after adjustment for age and height (p<0.05). CONCLUSIONS: These results indicate that, in this relatively lean population of Chinese children and adolescents, BP, particularly SBP, is positively associated with measures of adiposity. Of all the adiposity measures, BMI is the strongest predictor of BP. PMID- 18381343 TI - Torticollis, facial asymmetry and plagiocephaly in normal newborns. AB - OBJECTIVE: To evaluate the incidence and characteristics of torticollis, plagiocephaly and facial asymmetry in normal newborn infants. DESIGN: 102 healthy newborn infants were examined prospectively during their birth hospitalisation for torticollis with neck range of motion (ROM) assessment and for facial, mandibular and cranial asymmetry by photographic analysis. RESULTS: 73% of newborns had one or more asymmetry: torticollis (16%), asymmetry of the mandible (13%), facial asymmetry (42%) and asymmetry of the head (61%). Torticollis was associated with maternal report of the fetus being "stuck" in one intrauterine position for more than 6 weeks before delivery. Moderate facial asymmetry was associated with a longer second stage of labour, forceps delivery, a bigger baby and birth trauma. Moderate cranial and mandibular asymmetries were associated with birth trauma. More than one significant asymmetry was found in 10% of newborns. CONCLUSIONS: Asymmetries of the head and neck are very common in normal newborns, and sixteen (16%) of 102 study newborns were found to have torticollis. Such newborns, especially if they sleep supine, are thought to be at risk of developing deformational posterior plagiocephaly. Identification of affected infants may allow early implementation of positioning recommendations or physical therapy to prevent the secondary craniofacial deformations that are part of an increasingly common phenomenon. PMID- 18381345 TI - Serious injuries in children: variation by area deprivation and settlement type. AB - OBJECTIVE: To examine the variation in serious injuries in children by the level of deprivation in an area and by types of settlement. METHODS: Hospital admission rates for serious injury to children aged 0-15 years in census lower super output areas in England during the 5-year period 1 April 1999 to 31 March 2004 were analysed. RESULTS: Rates of serious injury in children were higher in the most deprived areas than in the least deprived for pedestrians (rate ratio (RR) 4.1; 95% CI 2.8 to 6.0) and cyclists (RR 3.0; 95% CI 1.9 to 4.7). Rates of serious pedestrian injury were lower in towns and fringe areas (RR 0.67; 95% CI 0.53 to 0.86) and in village (RR 0.64; 95% CI 0.50 to 0.83) areas than in urban areas. The rate of serious injury to cyclists was lower in London than other urban areas (RR 0.78; 95% CI 0.62 to 0.98). The rate of serious injury to car occupants was higher in village than urban areas (RR 1.51; 95% CI 1.05 to 2.17). Rates of serious injury caused by falls were higher in London (RR 1.60; 95% CI 1.47 to 1.75) and lower in villages (RR 0.76; 95% CI 0.66 to 0.88) than in urban areas. Steeper socio-economic gradients in serious injury rates were identified in rural areas for cyclists and for children suffering falls. CONCLUSIONS: Socio-economic inequalities in serious injury exist across the whole of England, particularly for child pedestrians. Rates of serious injury vary by settlement type, and inequalities vary by cause of injury between rural and urban settings. PMID- 18381346 TI - The management of pulmonary hypertension in children. AB - Pulmonary hypertension is relatively common in children and has many causes. The management of the condition has changed dramatically in the past 5 years with the introduction of new medicines. However, diagnosis, investigation and choice of therapy remain a challenge. In 2002 the United Kingdom Pulmonary Hypertension Service for Children was established and this has become the mainstay of management in this country. This service, based at Great Ormond Street Hospital for Children, provides advice, expertise and infrastructure support for the most severely affected patients, particularly those with idiopathic pulmonary arterial hypertension for whom chronic intravenous prostacyclin remains the most effective medication. New medicines are being developed which, rather than focussing on dilating a diseased pulmonary vascular bed, aim to structurally remodel the pulmonary vasculature towards normal. PMID- 18381347 TI - Improving patient safety in paediatrics and child health. PMID- 18381348 TI - Weight gain with pizotifen therapy. AB - OBJECTIVE: To describe the characteristics of weight gain occurring in children treated for migranous conditions with pizotifen DESIGN: Retrospective case note review SETTING: West Suffolk Hospital between 1999 and 2003 PATIENTS: 405 clinic attendances of 121 children receiving pizotifen treatment MAIN OUTCOME MEASURES: Rates of change in weight for age z score between successive outpatient appointments were used to detect excessive weight gain. RESULTS: The mean rate of weight z score increase for 105 patients receiving a constant dosage in 181 intervals between appointments was 0.79 standard deviations per year compared with 0.11 standard deviations per year for height z score increases. The rate of weight z score increase was not correlated with pizotifen dose per kg (r = 0.08), per m(2) (r = -0.05) or initial body mass index (r = -0.04). Mean rates of z score increase were similar in patients with a satisfactory and unsatisfactory therapeutic responses. CONCLUSIONS: Excess weight gain in pizotifen therapy is not predicted by drug dosage or therapeutic response within the range of doses used in clinical practice. PMID- 18381349 TI - Genetic interaction between Bardet-Biedl syndrome genes and implications for limb patterning. AB - Bardet-Biedl syndrome (BBS) is a pleiotropic, genetically heterogeneous disorder characterized by obesity, retinopathy, polydactyly, cognitive impairment, renal and cardiac anomalies, as well as hypertension and diabetes. Multiple genes are known to independently cause BBS. These genes do not appear to code for the same functional category of proteins; yet, mutation of each results in a similar phenotype. Gene knockdown of different BBS genes in zebrafish shows strikingly overlapping phenotypes including defective melanosome transport and disruption of the ciliated Kupffer's vesicle. Here, we demonstrate that individual knockdown of bbs1 and bbs3 results in the same prototypical phenotypes as reported previously for other BBS genes. We utilize the zebrafish system to comprehensively determine whether simultaneous pair-wise knockdown of BBS genes reveals genetic interactions between BBS genes. Using this approach, we demonstrate eight genetic interactions between a subset of BBS genes. The synergistic relationships between distinct combinations are not due to functional redundancy but indicate specific interactions within a multi-subunit BBS complex. In addition, we utilize the zebrafish model system to investigate limb development. Human polydactyly is a cardinal feature of BBS not reproduced in BBS-mouse models. We evaluated zebrafish fin bud patterning and observed altered Sonic hedgehog (shh) expression and subsequent changes to fin skeletal elements. The SHH fin bud phenotype was also used to confirm specific genetic interactions between BBS genes. This study reveals an in vivo requirement for BBS function in limb bud patterning. Our results provide important new insights into the mechanism and biological significance of BBS. PMID- 18381350 TI - Delta-like 1 is essential for the maintenance of marginal zone B cells in normal mice but not in autoimmune mice. AB - Notch2 and Delta-like 1 (Dll1) have been implicated in the development of marginal zone B (MZB) cells. In the present study, we characterized the expression and function of mouse Notch receptors and ligands in the spleen by using newly generated mAbs. Although Notch2 was expressed on both B and T cells in the spleen, the highest expression was observed on precursors of marginal zone B and MZB cells. Dll1 was expressed on macrophage and erythroblasts in the red pulp, but not on B cells or marginal zone macrophage. Administration of a blocking mAb against Dll1 not only blocked the development of MZB cells in juvenile mice but also gradually depleted the pre-established MZB cells in adult mice, indicating a critical role for Dll1 in the maintenance of MZB cells in the spleen of normal mice. Interestingly, Dll1 was not necessary for the maintenance of MZB cells in lupus-prone (NZB x NZW) F1 mice particularly after the onset of the disease, suggesting that the Dll1 independence may be a feature of dysregulated MZB cells producing auto-antibodies. PMID- 18381351 TI - Suppressor of cytokine signaling-1 ameliorates dextran sulfate sodium-induced colitis in mice. AB - Inflammatory bowel disease (IBD) is a chronic disorder of the gastrointestinal tract. Although the etiology and pathogenesis of IBD remain unknown, pro inflammatory cytokines including IFN-gamma play an important role in the development of IBD. Suppressor of cytokine signaling-1 (SOCS-1) is a crucial inhibitor of cytokine signaling, particularly of IFN-gamma. In this study, we investigated the role of SOCS-1 in the development of murine dextran sulfate sodium (DSS)-induced colitis, a model of colitis resembling human IBD. SOCS-1 heterozygous (SOCS-1(+/-)) and wild-type (WT) mice were given 3% DSS dissolved in drinking water for 5 days. Activation and expression of signal transducers and activators of transcription (STAT) in colonic tissues were assessed by western blot analysis. The expression of CD4, IFN-gamma, IL-4, IL-17 and Forkhead box P3 (Foxp3) in colonic lamina propria lymphocytes was analyzed by flow cytometry and cytokine concentrations in serum were measured. DSS-treated SOCS-1(+/-) mice developed more severe colitis than DSS-treated WT mice. Enhanced activation of STAT1, a higher ratio of CD4(+)IFN-gamma(+) T cells and a lower frequency of Foxp3(+) regulatory T (Treg) cells, were observed in the colon of DSS-treated SOCS-1(+/-) mice compared with DSS-treated WT mice. DSS-treated SOCS-1(+/-) mice showed higher levels of IFN-gamma in sera than did DSS-treated WT mice. Furthermore, T cell-specific SOCS-1-conditional knockout mice developed more severe colitis than control mice after DSS administration. Our findings suggest that SOCS-1, particularly in T cells, prevents the development of DSS-induced colitis in mice by inhibiting IFN-gamma/STAT1 signaling and by subsequently regulating Treg cell development. PMID- 18381352 TI - Strain distribution pattern of immune nephritis--a follow-up study. AB - Previous studies have indicated that the NZW, DBA/1, 129/sv and BUB strains are particularly sensitive to experimental anti-glomerular basement membrane (GBM) induced immune nephritis. The present study extends previous observations by examining eight additional inbred mouse strains for their susceptibility to immune nephritis. Unlike the ALR/Lt, CAST/Ei, DDY/JclSidSeyFrk, FVB/NJ, PERA/Ei, SB/Le and BALB/c strains, the C58 mouse strain was observed to be particularly susceptible to experimental immune nephritis, with CBA mice being a close second. In contrast to the other strains, C58 mice uniformly developed heavy proteinuria, azotemia and severe glomerulonephritis with prominent crescent formation and tubulointerstitial nephritis following challenge with anti-GBM sera. These differences were associated with increased murine Ig deposition, leukocyte infiltration and IFN-gamma production within the kidneys of C58 mice. Studies aimed at elucidating the genetic factors and molecular pathways responsible for the enhanced renal disease in C58 mice are warranted. PMID- 18381353 TI - A new mutant of Arabidopsis disturbed in its roots, right-handed slanting, and gravitropism defines a gene that encodes a heat-shock factor. AB - A new mutant of Arabidopsis named rha1 is characterized and the gene involved cloned. In roots, the mutant shows minimal right-handed slanting, reduced gravitropic response, notable resistance to 2,4-D, but scarce resistance to IAA and NAA. The roots also show a clear resistance to the auxin transport inhibitors TIBA and NPA, and to ethylene. Other characteristics are a reduced number of lateral roots and reduced size of shoot and root in the seedlings. The gene, cloned through TAIL-PCR, was found to be a heat-shock factor that maps on chromosome 5, close to and above the RFLP marker m61. The rha1 structure, mRNA, and translation product are reported. Since, so far, no other gravitropic mutant has been described as mutated in a heat-shock factor, rha1 belongs to a new group of mutants disturbed in slanting, gravitropism, and auxin physiology. As shown through the RT-PCR analyses of its expression, the gene retains the function connected with heat shock. If the characteristics connected with auxin physiology are considered, however, it is also likely that the gene, as a transcription factor, could be involved in root circumnutation, gravitropic response, and hormonal control of differentiation. Since GUS staining under the gene promoter was localized mainly in the mature tissues, rha1 does not seem to be involved in the first steps of gravitropism, but is rather related to the general response to auxin. The alterations in slanting (mainly due to reduced chiral circumnutation) and gravitropism lead to the supposition that the two processes may have, at least in part, common origins. PMID- 18381354 TI - Effect of PBPK model structure on interpretation of in vivo human aqueous dermal exposure trials. AB - Multiple research teams have reported data from in vivo human trials in which breath was monitored during and after whole-body or partial immersion in aqueous solutions of volatile organic compounds. Estimation of total dermal absorption from exhaled breath measurements requires modeling, a task to which physiologically based pharmacokinetic (PBPK) models have often been applied. In the context of PBPK models, the exposed skin compartment can be modeled in many different ways. To demonstrate potential effects of alternative skin models on overall PBPK model performance, alternative models of skin have been incorporated in a PBPK model used to predict chloroform in breath during and after immersion in aqueous solution. The models investigated include treatment of skin as both a homogeneous phase and as a membrane in which concentration varies with depth. Model predictions are compared with in vivo human experimental results reported in the prior literature. In the example chosen, the common practice of modeling skin as a homogenous phase leads to prediction of more rapid initial uptake and lower cumulative uptake than does modeling skin as a membrane. Numerical estimates of the permeability coefficient are shown to be dependent upon skin model form and temperature of the aqueous solution. PMID- 18381356 TI - Ecotoxicological applications and significance of the comet assay. AB - Application of the single-cell gel electrophoresis or comet assay has revolutionized the field of genetic ecotoxicology or eco-genotoxicology. It is a rapid, sensitive and relatively inexpensive method providing the opportunity to study DNA damage (including oxidative damage), repair and cell death (apoptosis) in different cell types without prior knowledge of karyotype and cell turnover rate. The assay has, however, often attracted criticism for its lack of ecotoxicological relevance. In addition, in contrast to genetic toxicology where rapid technical progress has been made to improve cell- and tissue-specific adoption of the assay, only limited advancement has been made to transfer the methodologies to ecotoxicological studies. While reviewing the recent information available in the literature and underscoring the importance of induced genetic damage in natural species, the aims of this article are to (i) highlight and judiciously analyse the ecotoxicological relevance of the assay; (ii) attempt to correlate the comet response with other relevant biological responses or biomarkers; (iii) identify the technical challenges and various factors affecting its application in order to make it reliable, reproducible and robust; (iv) critically compare the technical developments in genetic toxicology and genetic ecotoxicology and (v) evaluate the future developments with respect to applications of the assay. It is suggested that while complementing other ecotoxicological parameters and further improving the methodologies, the comet assay will continue to play an important role in genetic ecotoxicology to determine induced genetic damage, which has significant consequences for short- and long-term survival of the natural or wild species. Information obtained through integrated studies using simultaneous applications of multiple biomarkers on different wild organisms could also provide an holistic dimension of toxicological impact of environmental contaminants for the protection of human health. PMID- 18381355 TI - Pregnane X receptor protects HepG2 cells from BaP-induced DNA damage. AB - Pregnane X receptor (PXR) is a nuclear receptor that coordinately regulates transcriptional expression of both phase I and phase II metabolizing enzymes. PXR plays an important role in the pharmacokinetics of a broad spectrum of endogenous and xenobiotic compounds and appears to have evolved in part to protect organisms from toxic xenobiotics. Metabolism of benzo[a]pyrene (BaP), a well-established carcinogen and ubiquitous environmental contaminant, can result in either detoxification or bioactivation to its genotoxic forms. Therefore, PXR could modulate the genotoxicity of BaP by changing the balance of the metabolic pathways in favor of BaP detoxification. To examine the role of PXR in BaP genotoxicity, BaP-DNA adduct formation was measured by 32P-postlabeling in BaP treated parental HepG2 cells and human PXR-transfected HepG2 cells. The presence of transfected PXR significantly reduced the level of adducts relative to parental cells by 50-65% (p < 0.001), demonstrating that PXR protects liver cells from genotoxicity induced by exposure to BaP. To analyze potential PXR-regulated detoxification pathways in liver cells, a panel of genes involved in phase I and phase II metabolism and excretion was surveyed with real-time quantitative reverse transcription PCR. The messenger RNA levels of CYP1A2, GSTA1, GSTA2, GSTM1, UGT1A6, and BCRP (ABCG2) were significantly higher in cells overexpressing PXR, independent of exposure to BaP. In addition, the total GST enzymatic activity, which favors the metabolic detoxification of BaP, was significantly increased by the presence of PXR (p < 0.001), independent of BaP exposure. Taken together, these results suggest that PXR plays an important role in protection against DNA damage by polycyclic aromatic hydrocarbons (PAHs) such as BaP, and that these protective effects may be through a coordinated regulation of genes involved in xenobiotic metabolism. PMID- 18381357 TI - Genetic and disorder-specific aspects of resting state EEG abnormalities in schizophrenia. AB - We evaluated whether abnormal frequency composition of the resting state electroencephalogram (EEG) in schizophrenia was associated with genetic liability for the disorder by studying first-degree biological relatives of schizophrenia patients. The study included a data-driven method for defining EEG frequency components and determined the specificity of resting state EEG frequency abnormalities by assessing schizophrenia patients, bipolar disorder patients, and relatives of both patient groups. Schizophrenia patients and their relatives, but not bipolar patients or their relatives, exhibited increased high-frequency activity (beta) providing evidence for disturbances in resting state brain activity being specific to genetic liability for schizophrenia. Schizophrenia patients exhibited augmented low-frequency EEG activity (delta, theta), while bipolar disorder patients and the 2 groups of relatives generally failed to manifest similar low-frequency EEG abnormalities. The Val(158)Met polymorphism for the catechol-O-methyl transferase (COMT) gene was most strongly associated with delta and theta activity in schizophrenia patients. Met homozygote schizophrenia patients exhibited augmented activity for the 2 low-frequency bands compared with control subjects. Excessive high-frequency EEG activity over frontal brain regions may serve as an endophenotype that reflects cortical expression of genetic vulnerability for schizophrenia. Low-frequency resting state EEG anomalies in schizophrenia may relate to disorder-specific pathophysiology in schizophrenia and the influence of the COMT gene on tonic dopamanergic function. PMID- 18381358 TI - Regulation of hypoxia-inducible genes by ETS1 transcription factor. AB - Hypoxia-inducible factor (HIF-1) regulates the expression of genes that facilitate tumor cell survival by making them more resistant to therapeutic intervention. Recent evidence suggests that the activation of other transcription factors, in cooperation with HIF-1 or acting alone, is involved in the upregulation of hypoxia-inducible genes. Here we report that high cell density, a condition that might mimic the physiologic situation in growing tumor and most probably representing nutritional starvation, upregulates hypoxia-inducible genes. This upregulation can occur in HIF-independent manner since hypoxia inducible genes carbonic anhydrase 9 (CA9), lysyloxidase like 2 (LOXL2) and n-myc down regulated 1 (NDRG1)/calcium activated protein (Cap43) can be upregulated by increased cell density under both normoxic and hypoxic conditions in both HIF-1 alpha-proficient and -deficient mouse fibroblasts. Moreover, cell density upregulates the same genes in 1HAEo- and A549 human lung epithelial cells. Searching for other transcription factors involved in the regulation of hypoxia inducible genes by cell density, we focused our attention on ETS1. As reported previously, members of v-ets erythroblastosis virus E26 oncogene homolog (ETS) family transcription factors participate in the upregulation of hypoxia-inducible genes. Here, we provide evidence that ETS1 protein is upregulated at high cell density in both human and mouse cells. The involvement of ETS1 in the upregulation of hypoxia-inducible genes was further confirmed in a luciferase reporter assay using cotransfection of ETS1 expression vector with NDRG1/Cap43 promoter construct. The downregulation of ETS1 expression with small interfering RNA (siRNA) inhibited the upregulation of CA9 and NDRG1/Cap43 caused by increased cell density. Collectively, our data indicate the involvement of ETS1 along with HIF-1 in regulating hypoxia-inducible genes. PMID- 18381359 TI - Differentially expressed nucleolar transforming growth factor-beta1 target (DENTT) exhibits an inhibitory role on tumorigenesis. AB - Differentially expressed nucleolar transforming growth factor-beta1 target (DENTT), also known as testis-specific protein Y-encoded-like (TSPYL-2) and cell division autoantigen-1, is a member of the testis-specific protein Y-encoded (TSPY)/TSPY-L/SET/nucleosome assembly protein-1 superfamily. DENTT is expressed in various tissues including normal human lung. Here, we investigate the involvement of DENTT in cancer promotion and progression. DENTT messenger RNA (mRNA) and protein levels were shown to be markedly downregulated in human and mouse primary tumors and in human tumor cell lines. Overexpression of DENTT in human lung (A549-DENTT) and breast (MCF-7-DENTT) cancer cells resulted in diminished growth potential in anchorage-dependent growth assays and reduced capacity to form colonies under anchorage-independent culture conditions. The migratory potential of A549-DENTT and MCF-7-DENTT cells was reduced when compared with empty vector control cells. Treating human lung cell lines with demethylating agents increased DENTT expression significantly. DENTT expression pattern paralleled that of transforming growth factor-beta1 (TGF-beta1) in normal and malignant tissue and ectopic expression or treatment with TGF-beta1 in lung cancer cells was followed by increased DENTT mRNA and protein levels. Collectively, our results suggest a role for DENTT as a suppressor of the tumorigenic phenotype. PMID- 18381360 TI - Trend of increase in the incidence of acute myocardial infarction in a Japanese population: Takashima AMI Registry, 1990-2001. AB - The incidence and mortality of acute myocardial infarction (AMI) remain low in Japan despite major dietary changes and worsening cardiovascular risk factors, a situation that should have resulted in a substantial increase in AMI rates (Japanese paradox). The current trend in the incidence of AMI was examined for the period 1990-2001 by use of data from the Takashima AMI Registry covering a stable population of approximately 55,000 in central Japan. AMI incidence rates (per 100,000 person-years) and 95% confidence intervals were calculated for 1990 1992, 1993-1995, 1996-1998, and 1999-2001. The incidence trend was determined by calculating the average annual change in percentage across the years. There were 352 (men: n = 224; women: n = 128) registered first-ever AMI cases during 1990 2001. The age-adjusted incidence rate of all AMI showed a gradual increase from 39.9 (95% confidence interval (CI): 29.8, 50.0) in 1990-1992 to 62.6 (95% CI: 51.5, 73.7) in 1999-2001. In men, the age-adjusted incidence rate increased from 66.5 (95% CI: 46.4, 86.6) in 1990-1992 to 100.7 (95% CI: 78.6, 122.7) in 1999 2001. In women, fluctuation was observed after an initial steep increase. The average annual incidence increased by 7.6% (95% CI: 3.5, 11.7) among men and by 8.3% (95% CI: 1.02, 15.6) among women. To the best of the authors' knowledge, this is the first study to report an increasing trend of AMI in a Japanese population. PMID- 18381361 TI - The role of muscle flap in preventing bronchus stump insufficiency after pneumonectomy for malignant pleural mesothelioma in high-risk patients. AB - Bronchus stump insufficiency (BSI) is one of the major complications after pneumonectomy; we analyzed all patients who underwent extra pleural pneumonectomy (EPP) for malignant pleural mesothelioma (MPM) in order to detect the role of muscle flap (MF) on preventing early and late stump insufficiency. From January 2000 until December 2005, there were 42 patients admitted with MPM for further intervention at our institution. Thirty patients were suitable for surgery and thus received a multimodal treatment with neo-adjuvant chemotherapy using Cisplatin and Gemcitabin (Gemzar), EPP followed by 54 Gray (Gy) adjuvant radiotherapy. Data were collected from the surgical and oncological records. There were 37 male patients (88%), the median age was 65 years (range 40-83 years). Seven (17%) patients had concomitant diseases. Forty patients (95%) had asbestos exposition. The operative procedures were EPP with muscle flap through an anterolateral thoracotomy. Univariate and multivariate analyses were done. One patient (3%) died on the 2nd postoperative day due to lung embolism. Mild complications were noticed in the early postoperative phase in 8 (25%) patients. There was no early or late stump insufficiency during the 15-month follow-up. Surgical techniques using muscle flap seems to play a major role in the prevention of bronchus stump insufficiency especially after neo-adjuvant chemotherapy. PMID- 18381362 TI - Carbon dioxide embolism during endoscopic vein harvesting. AB - Endoscopic vein harvesting (EVH) is becoming common for the patients undergoing coronary artery bypass grafting. Using carbon dioxide insufflations during the vein harvest can produce rare but catastrophic CO(2) embolism. We report a case of massive right atrial CO(2) embolism due to femoral vein injury which occurred during the performance of a routine EVH procedure. PMID- 18381363 TI - eComment: Long saphenous vein harvesting and recurrences. PMID- 18381364 TI - eComment: The anastomosis between aorta and extension conduit of the pulmonary artery. PMID- 18381365 TI - eComment: Bilateral atrial appendage excision should be performed routinely in the surgical treatment of atrial fibrillation. PMID- 18381366 TI - eComment: Foreign body reaction to polytetrafluoroethylene. PMID- 18381367 TI - eComment: TEE- and guidewire-guided axillary artery cannulation. An option? PMID- 18381368 TI - eComment: Can we use endovascular graft stenting in all type B dissections? PMID- 18381369 TI - Effects of an endurance and resistance exercise program on persistent cancer related fatigue after treatment. AB - BACKGROUND: Fatigue is a relevant problem of cancer patients during and after treatment. Several studies have shown that exercise can improve quality of life and functional status of cancer patients undergoing chemo- or radiotherapy. However, there is a lack of information about the effects of this intervention on persistent cancer-related fatigue. Therefore, we assessed the effects of an exercise program on cancer-related fatigue after treatment. PATIENTS AND METHODS: A consecutive series of 32 cancer patients with mild to severe persistent fatigue [scores on the Brief Fatigue Inventory (BFI) > 25] participated in a 3-week exercise program consisting of endurance (30 min walking on a treadmill) and resistance/coordination exercises for the major muscle groups. Fatigue, mood, and anxiety were assessed with questionnaires and physical performance with a stress test before and after the program. RESULTS: At the end of the program, we observed a significant increase of physical performance (workload at the anaerobic threshold pre 61 +/- 26 W, post 78 +/- 31 W, P < 0.0001) and reduction of global fatigue (Functional Assessment of Cancer Therapy: pre 45.7 +/- 13.4, post 52.6 +/- 12.4, P < 0.0001; BFI: pre 37.9 +/- 18.3, post 31.2 +/-17.1, P < 0.001). However, no significant improvement of cognitive fatigue or reduction of anxiety was observed. CONCLUSIONS: A 3-week exercise program leads to a substantial improvement of physical performance and reduction of mental and physical fatigue in cancer patients after treatment. However, this intervention does not affect depression, anxiety, or cognitive fatigue. PMID- 18381370 TI - UFT (tegafur-uracil) in rectal cancer. AB - BACKGROUND: Major achievements in the treatment of localised rectal cancer include the development of total mesorectal excision and the perioperative administration of radiotherapy in combination with continuous infusion (CI) 5 fluorouracil (5-FU). This multimodal approach has resulted in extended survival and lower local relapse rates, with the potential for sphincter-preserving procedures. However, CI 5-FU is inconvenient for patients and is costly. Oral fluoropyrimidines like UFT (tegafur-uracil) offer a number of advantages over 5 FU. METHODS: We undertook a review of published articles and abstracts relating to clinical studies of UFT in the treatment of locally advanced rectal cancer (LARC). Pre- and postoperative studies carried out in patients with newly diagnosed or recurrent disease were included. RESULTS: The combination of UFT and radiotherapy was effective and well tolerated in the preoperative setting, while adjuvant UFT improved survival and reduced distant relapse compared with surgery alone. The efficacy of UFT appears comparable with that of 5-FU and capecitabine and its side-effect profile is favourable. CONCLUSION: Clinical experience to date suggests that UFT is a valuable treatment option for the perioperative treatment of LARC. Further improvements in patient outcomes may result from the combination of UFT with targeted agents. PMID- 18381371 TI - Pancreatic cancer--is the wall crumbling? AB - In spite of advances made in the management of the other more common cancers of the gastrointestinal tract, significant progress in the treatment of pancreatic cancer remains elusive, more so with the recent negative results of several much anticipated randomized trials. Gemcitabine has been a standard treatment for advanced pancreatic cancer since it was shown a decade ago to result in a superior clinical benefit response and survival compared with bolus 5 fluorouracil (5-FU). Since then, clinical trials have explored the pharmacokinetic modulation of gemcitabine by fixed dose administration and the combination of gemcitabine with other cytotoxics or the biological 'targeted' agents. Against a background of numerous negative randomized trials of gemcitabine-based combination treatment, two trials have recently reported modest survival improvements with the use of combination treatment: the United Kingdom National Cancer Research GEMCAP trial of gemcitabine with the orally administered precursor of 5-FU-capecitabine and the National Cancer Institute of Canada Clinical Trials Group PA.3 trial in which the tyrosine kinase inhibitor erlotinib was used with gemcitabine. This review will summarize the results of several recent randomized trials of combination treatment in advanced pancreatic cancer and discuss their implications for clinical practice and for future research in this disease. PMID- 18381372 TI - Phase II multicenter study of larotaxel (XRP9881), a novel taxoid, in patients with metastatic breast cancer who previously received taxane-based therapy. AB - BACKGROUND: Treatment options are limited for patients with refractory metastatic breast cancer (MBC). Larotaxel (XRP9881) is a novel taxoid with preclinical activity against taxane-resistant breast cancer. The current phase II trial of larotaxel was conducted in women with taxane-treated MBC. PATIENTS AND METHODS: Patients were stratified by response to prior taxane therapy (resistant or nonresistant). Larotaxel 90 mg/m(2) was administered as a 1-h infusion every 3 weeks. Patients were evaluated for tumor response every two cycles. A blinded external response review committee determined the overall response rate (ORR), duration of response (DOR), and time to progression (TtP) of the disease. Median survival time (MST) and safety were also evaluated. RESULTS: One hundred and thirty patients were treated. In the nonresistant group, the ORR was 42%; median DOR 5.3 months; median TtP 5.4 months; and MST 22.6 months. In the resistant group, the ORR was 19%; median DOR 5.0 months; median TtP 1.6 months; and MST 9.8 months. The most common grade 3/4 adverse events were neutropenia (82%), fatigue (15%), diarrhea (12%), febrile neutropenia (9%), neutropenic infection (8%), and sensory neuropathy (7%). CONCLUSIONS: Larotaxel has good activity, manageable toxicity, and a favorable therapeutic index in women with taxane-pretreated MBC. PMID- 18381373 TI - Prognostic and therapeutic implications of dipyridamole stress cardiovascular magnetic resonance on the basis of the ischaemic cascade. AB - OBJECTIVE: To determine the prognostic and therapeutic implications of stress perfusion cardiovascular magnetic resonance (CMR) on the basis of the ischaemic cascade. SETTING: Single centre study in a teaching hospital in Spain. PATIENTS: Dipyridamole stress CMR was performed on 601 patients with ischaemic chest pain and known or suspected coronary artery disease. On the basis of the ischaemic cascade, patients were categorised in C1 (no evidence of ischaemia, n = 354), C2 (isolated perfusion deficit at stress first-pass perfusion imaging, n = 181) and C3 (simultaneous perfusion deficit and inducible wall motion abnormalities, n = 66). CMR-related revascularisation (n = 102, 17%) was defined as the procedure prompted by the CMR results and carried out within the next three months. RESULTS: During a median follow-up of 553 days, 69 major adverse cardiac events (MACE), including 21 cardiac deaths, 14 non-fatal myocardial infarctions and 34 admissions for unstable angina with documented abnormal angiography were detected. In non-revascularised patients (n = 499), the MACE rate was 4% (14/340) in C1, 20% (26/128) in C2 and 39% (12/31) in C3 (adjusted p value = 0.004 vs C2 and <0.001 vs C1). CMR-related revascularisation had neutral effects in C2 (20% vs 19%, 1.1 (0.5 to 2.4), p = 0.7) but independently reduced the risk of MACE in C3 (39% vs 11%, 0.2 (0.1 to 0.7), p = 0.01). CONCLUSIONS: Dypiridamole stress CMR is able to stratify risk on the basis of the ischaemic cascade. A small group of patients with severe ischaemia-simultaneous perfusion deficit and inducible wall motion abnormalities-are at the highest risk and benefit most from MACE reduction due to revascularisation. PMID- 18381374 TI - Should the first priority in cardiovascular risk management be those with prior cardiovascular disease? AB - BACKGROUND: Cardiovascular disease (CVD) prevention guidelines typically dichotomise patients by history of CVD, as patients with prior CVD are assumed to be at high CVD risk, whatever their CVD risk profiles. OBJECTIVE: To assess the appropriateness of this practice by comparing CVD event rates of patients with and without prior CVD, over and above risk predicted by standard CVD risk factors. METHODS: Between 2002 and 2007 CVD risk assessments were generated using a web-based Framingham risk prediction algorithm in routine primary care. Individual risk profiles were subsequently linked to national hospitalisation and death records. Observed and predicted (Framingham) CVD risk were compared in patients with and without prior CVD. RESULTS: 35 760 patients were assessed including 10.4% with prior CVD. Of 1216 first CVD events during an average follow up of 2.05 years, 42% occurred in those with prior CVD. Among those without prior CVD, the predicted Framingham five-year CVD risk was similar to the observed risk extrapolated to five years; in the highest Framingham risk band (>20% five-year risk), observed risk was 25.3%. Among those with prior CVD the observed risk extrapolated to five years rose from 21.7% in the lowest Framingham risk band (<5%) to 49% in the highest (>20%). CONCLUSIONS: Patients with prior CVD have five-year CVD risks approximately 20% higher, in absolute terms than patients without prior CVD, after accounting for standard risk factors. Almost half the CVD events occurred in those with prior CVD. These patients should be the highest priority for intensive preventive management in primary care. PMID- 18381375 TI - Acute hyperglycaemia prevents the protective effect of pre-infarction angina on microvascular function after primary angioplasty for acute myocardial infarction. AB - BACKGROUND: Acute hyperglycaemia has been associated with impaired microvascular function after acute myocardial infarction (AMI), whereas pre-infarction angina (PIA) occurring shortly before the onset of AMI has been shown to reduce microvascular injury after reperfusion. OBJECTIVE: To examine whether acute hyperglycaemia prevents the protective effect of PIA on microvascular function after AMI. METHODS: We studied 205 patients with a first anterior wall AMI who underwent primary angioplasty within 12 hours of onset. Coronary flow velocity parameters were assessed immediately after reperfusion using a Doppler guidewire. Severe microvascular injury was defined as the presence of systolic flow reversal and diastolic deceleration time <600 ms. Echocardiographic wall motion was analysed before revascularisation and 4 weeks later. RESULTS: Acute hyperglycaemia, defined as a blood glucose level of >or=198 mg/dl on admission, was found in 67 (33%) patients. In patients without acute hyperglycaemia, PIA was associated with a lower incidence of systolic flow reversal, a longer diastolic deceleration time and a higher coronary flow reserve. However, in patients with acute hyperglycaemia there was no significant difference in these same parameters between patients with and without PIA. In the presence of acute hyperglycaemia PIA did not improve the change in wall motion score. In a multivariate model, the absence of PIA was an independent determinant of severe microvascular injury in patients without acute hyperglycaemia (odds ratio 6.28, p = 0.001), but not in patients with acute hyperglycaemia. CONCLUSION: The protective effect of PIA on microvascular function was attenuated in patients with acute hyperglycaemia, resulting in unfavourable functional recovery. PMID- 18381376 TI - Predictors of slow flow during primary percutaneous coronary intervention: an intravascular ultrasound-virtual histology study. AB - OBJECTIVES: Slow flow phenomenon is a serious complication of percutaneous coronary intervention (PCI) and is associated with a poor prognosis. We sought to evaluate the characteristics of lesions predisposing to the slow/no-reflow phenomenon during primary PCI in patients with acute myocardial infarction. METHODS: The study subjects consisted of 57 consecutive patients (mean age 58.5 (SD 14.5) years, 45 males) who underwent primary PCI for acute myocardial infarction and intravascular ultrasound-virtual histology (IVUS-VH) examination. Slow flow was defined as 0.55 cm(2)/m(2)). RESULTS: Compared with healthy controls (n = 14), patients with low-flow, low-gradient AS had higher resting mean (SD) MBF (0.83 (0.21) vs 0.69 (0.09) ml/min/g, p = 0.001), reduced hyperaemic MBF (1.16 (0.31) vs 2.71 (0.50) ml/min/g, p<0.001) and impaired MFR (1.44 (0.44) vs 4.00 (0.91), p<0.001). Resting MBF and MFR correlated with indices of AS severity in low-flow, low-gradient AS with the strongest relationship observed for EOAI(proj) (r(s) = -0.50, p = 0.002 and r(s) = 0.61, p = 0.004, respectively). Compared with PSAS, TSAS had a trend to a higher resting MBF (0.90 (0.19) vs 0.77 (0.21) ml/min/g, p = 0.06), similar hyperaemic MBF (1.16 (0.31) vs 1.17 (0.32) ml/min/g, p = NS), but a significantly smaller MFR (1.19 (0.26) vs 1.76 (0.41), p = 0.003). An MFR <1.8 had an accuracy of 85% for distinguishing TSAS from PSAS. CONCLUSIONS: Low-flow, low-gradient AS is characterised by higher resting MBF and reduced MFR that relates to the AS severity. The degree of MFR impairment differs between TSAS and PSAS and may be of value for distinguishing these entities. PMID- 18381379 TI - The combined oxacillin resistance and coagulase (CORC) test for rapid identification and prediction of oxacillin resistance in Staphylococcus species directly from blood culture. AB - The combined oxacillin resistance and coagulase (CORC) protocol for rapid identification and determination of oxacillin-susceptibility in Staphylococcus spp from blood culture is described. It incorporates a modified direct tube coagulase test (TCT) and a novel 4-hour multiplication-induction step, which increases the expression of staphylococcal PBP2a if present, facilitating detection by a commercial PBP2a latex agglutination kit. The protocol shows excellent sensitivity and specificity for determination of coagulase-positivity in staphylococci from patient blood cultures (96.8% (95% CI 81.5 to 99.8) and 100% (95% CI 75.9 to 100), respectively, n = 47), and for prediction of oxacillin resistance in S aureus directly from patient blood cultures (100% (95% CI 59.8 to 100) and 100% (95% CI 82.2 to 100), respectively (100% accuracy), n = 31) within 5 hours of blood culture positivity. PMID- 18381380 TI - HER2 testing in the UK: further update to recommendations. AB - These guidelines update the previous UK HER2 testing guidelines and have been formulated to give advice on methodology, interpretation and quality assurance to ensure that HER2 testing results are accurate, reliable and timely with the expansion of testing to all patients with breast cancer at the time of primary diagnosis. The recommendations for testing are the use of immunohistochemistry but with analysis of equivocal cases by in situ hybridisation to clarify their HER2 status or the use of frontline fluorescence in situ hybridisation (FISH) testing for those laboratories wishing to do so; the inclusion of a chromosome 17 probe is strongly recommended. Laboratories using chromogenic or silver in situ hybridisation should perform an initial validation against FISH. For immunohistochemistry and in situ hybridisation there must be participation in the appropriate National External Quality Assurance scheme. PMID- 18381381 TI - Breast carcinomas that co-express E- and P-cadherin are associated with p120 catenin cytoplasmic localisation and poor patient survival. AB - BACKGROUND: Changes in junctional catenin expression may compromise cadherin mediated adhesion, increasing cell malignant properties such as invasive and metastatic abilities. Altered expression of alpha-, beta-, gamma- and p120 catenin has been reported to be associated with E-cadherin loss or decreased expression, in both breast carcinomas and breast cancer cell lines. AIMS AND METHODS: To investigate the expression and subcellular localisation of p120- and beta-catenin in a series of human invasive breast carcinomas, and correlate it with biological markers and clinicopathological parameters. RESULTS: Both catenins frequently exhibited a reduced membranous or cytoplasmic staining pattern. These alterations were significantly correlated with lack of both E cadherin and oestrogen receptor-alpha expression. It was possible to associate the expression of beta-catenin with histological grade, tumour size and nodal status, suggesting a relevant role for this catenin as a prognostic factor. The majority of E- and P-cadherin co-expressing tumours were related to cytoplasmic expression of p120-catenin; in this group of breast carcinomas, patient survival was poor. CONCLUSION: Results indicate that p120-catenin cytoplasmic accumulation may play an important role in mediating the oncogenic effects derived from P cadherin aberrant expression, including enhanced motility and invasion, particularly in tumours which maintain E-cadherin expression. PMID- 18381382 TI - Neurologically presenting Whipple disease: case report and review of the literature. AB - A previously healthy male with subacute onset right leg weakness was suspected to have an astrocytoma as imaging showed a lesion. Subsequent biopsy showed the presence of foamy macrophages containing periodic acid-Schiff staining granules, suggesting Whipple disease as a possible diagnosis. PMID- 18381383 TI - Characteristics of KIT-negative gastrointestinal stromal tumours and diagnostic utility of protein kinase C theta immunostaining. AB - AIMS: To characterise KIT-negative gastrointestinal stromal tumours (GISTs) clinically, pathologically, immunohistochemically and genetically, and to establish the usefulness of protein kinase C theta (PKC) as a diagnostic marker in KIT-negative GIST. METHODS: 252 consecutive cases of GIST were evaluated for clinicopathological characteristics and immunostained for various antibodies. Mutational analyses of KIT and platelet-derived growth factor receptor alpha (PDGFRA) were also performed in 62 cases. RESULTS: 20 (7.9%) GISTs showed negative immunostaining for KIT. KIT-negative GISTs were more likely to originate from omentum or peritoneum, have an epithelioid histology, and be classified as high risk. The overall survival rate of patients with KIT-negative GISTs (5-year survival rate 68.7% (SD 10.7%)) was lower than that of patients with KIT-positive GISTs (5-year survival rate, 79.9% (3.0%)) (p = 0.042, log-rank test). Negative KIT expression was an independent prognostic factor in multivariate Cox regression analysis when the risk of aggressive behaviour and the status of imatinib treatment were adopted as covariates. KIT-negative GISTs also showed lower expression rates of CD34, Bcl-2, and PKC than KIT-positive GISTs; mutational analysis revealed that 30% of KIT-negative GISTs harboured a PDGFRA exon 18 mutation. Immunostaining on PKC showed that 93.9% of all GISTs expressed PKC protein. However, 21.9% of 64 mesenchymal tumours other than GIST also showed positivity on PKC. CONCLUSIONS: KIT-negative GISTs had characteristics that differ from those of KIT-positive GISTs, and negative KIT expression was an independent prognostic indicator for overall survival of patients. Although PKC is a sensitive diagnostic marker for GIST, its usefulness is limited because of low sensitivity and low specificity in KIT-negative GISTs. PMID- 18381384 TI - Pandemic influenza preparedness in Africa is a profound challenge for an already distressed region: analysis of national preparedness plans. AB - A new highly pathogenic strain of influenza virus, H5N1, has emerged causing severe outbreaks in poultry and high mortality rates when humans are infected. The threat of a new influenza pandemic has prompted countries to draft national strategic preparedness plans to prevent, contain and mitigate the next human influenza pandemic. To evaluate preparedness for an influenza pandemic in the African region we analysed African national preparedness plans available in the public domain. A data extraction tool, based on a World Health Organization checklist for influenza epidemic preparedness, was designed in consultation with pandemic influenza planning experts and experts on the region's public health challenges. Thirty-five plans were identified and available from 53 African countries. Most plans are relatively robust in addressing detection and containment of influenza in animals but strategic preparedness to respond to pandemic human influenza is weak. In most plans communication strategies have been developed with the aim to raise awareness of transmission factors and promote hygiene measures. By contrast, the human health care sector is ill prepared. Case management, triage procedures, identification of health care facilities for patient treatment (including home care and provisions for the distribution and administration of pharmaceuticals) are poorly addressed by most plans. The maintenance of essential services in the event of a pandemic is absent from most plans. Whilst many African countries have strategic pandemic influenza preparedness plans, most are developmental in nature and lack operational clarity, or focus principally on the containment of avian influenza rather than pandemic human influenza. Clear strategies, that are operational, need to be developed that reflect the realities of national context and resource constraints and that meet national objectives. These objectives need also to be coherent with international imperatives such that the global threat of pandemic influenza can be met effectively and efficiently. PMID- 18381385 TI - Central nervous system Aspergillus fumigatus presenting as cranial nerve palsy in a patient with ankylosing spondylitis on anti-TNF therapy. PMID- 18381387 TI - Levels of 5' RNA tags in plasma and buffy coat from EDTA blood increase with time. AB - BACKGROUND: For biological sample banking it is important to precisely document sample treatment prior to extraction and storage. A major variable is the interval between blood sampling and subsequent processing and storage. We have determined the relationship between this time interval and frequency of 5' transcript tags. This study was designed to establish guidelines for collecting RNA from blood in prospective studies and ensure maximum availability of RNA analytes. METHODS: Venous blood was collected from 40 healthy volunteers. Samples were processed immediately, 12, 24 and 36 h post collection and buffy coat and/or plasma removed. Total RNA was extracted and reverse transcribed, assays were optimized and levels of 5' RNA tags quantified by qPCR. RESULTS: Stably expressed reference genes were selected to examine 5' tags in plasma and buffy coat blood fractions. Whole blood was processed at various time points post collection to determine the affect on the presence and stability of 5' RNA tags. A significant increase (P < 0.05 to P < 0.001) in 5' RNA tags was observed at 12 h and up to 36 h in plasma and buffy coat samples isolated from EDTA blood which was maintained at 4 degrees C prior to processing when compared with plasma and buffy coat isolated from EDTA blood processed immediately. CONCLUSIONS: Over time 5' RNA tags increase in both plasma and buffy coat samples. It has been previously shown that removing cells from their normal environment produces cellular activation and up-regulation of pathways resulting in increased transcript expression. Positive correlation was observed between the time interval from sample collection to storage and amount of 5' transcript tags present. This increase could be due to white blood cells undergoing necrosis and lysis, or from RNA protected within apoptotic bodies. As 5' RNA tags were targeted using random primers for reverse transcription, even RNA partly degraded by RNases would have been detected. PMID- 18381388 TI - UK Biobank Pilot Study: stability of haematological and clinical chemistry analytes. AB - BACKGROUND: Analytes in blood and urine may vary over time according to conditions of transport and storage. METHODS: UK Biobank pilot study to investigate stability through time of 42 haematological and clinical chemistry analytes in blood and four analytes in urine, kept in storage for up to 36 h, for 40 individuals. Random effects linear regressions were used to model the change through time in repeated assay results on a sample, allowing for heterogeneity between individuals and assay variability. RESULTS: Assay results for most analytes tended to show a small negative bias (1-3% per 12 h stored) over time on average. Statistically significant (P < 0.05) heterogeneity in time trends between individuals, found for nearly all analytes, was dominated by differences in the baseline (time 0) assay results with the possible exception of Mean Corpuscular Haemoglobin Concentration (MCHC). Four out of 46 analytes (serum calcium, cholesterol, fibrinogen and HDL cholesterol) had a predicted probability of a negative time trend for a future individual >0.9. Results for freeze-thaw samples were not materially different from those for non-freeze-thaw samples, except that stability of the analyte results was only assessed up to 12 h. CONCLUSIONS: The results suggest that any instability in assay results up to 36 h is likely to be small in comparison with between individual differences and assay error, and that a single assay measurement at any time between 0 and 36 h should give a representative value of the analyte concentration at time zero for that individual. PMID- 18381389 TI - The UK Biobank sample handling and storage validation studies. AB - BACKGROUND: and aims UK Biobank is a large prospective study in the United Kingdom to investigate the role of genetic factors, environmental exposures and lifestyle in the causes of major diseases of late and middle age. It involves the collection of blood and urine from 500 000 individuals aged between 40 and 69 years. How the samples are collected, processed and stored will have a major impact on the future scientific usefulness of the UK Biobank resource. A series of validation studies was recommended to test the robustness of the draft sample handling and storage protocol. METHODS: Samples of blood and urine were collected from 40 healthy volunteers and either processed immediately according to the protocol or maintained at specified temperatures (4 degrees C for all tubes with the exception of vacutainers containing acid citrate dextrose that were maintained at 18 degrees C) for 12, 24 or 36 h prior to processing. A further sample was maintained for 24 h at 4 degrees C, processed and the aliquots frozen at -80 degrees C for 20 days and then thawed under controlled conditions. The stability of the samples was compared for the different times in a wide variety of assays. RESULTS: The samples maintained at 4 degrees C were stable for at least 24 h after collection for a wide range of assays. Small but significant changes were observed in metabonomic studies in samples maintained at 4 degrees C for 36 h. There was no degradation of the samples for a range of biochemical assays after short-term freezing and thawing under controlled conditions. Whole blood maintained at 18 degrees C for 24 h in vacutainers containing acid citrate dextrose is suitable for viral immortalization techniques. CONCLUSIONS: The validation studies reported in this supplement provide justification for the sample handling and storage procedures adopted in the UK Biobank project. PMID- 18381390 TI - A GC-TOF-MS study of the stability of serum and urine metabolomes during the UK Biobank sample collection and preparation protocols. AB - BACKGROUND: The stability of mammalian serum and urine in large metabolomic investigations is essential for accurate, valid and reproducible studies. The stability of mammalian serum and urine, either processed immediately by freezing at -80 degrees C or stored at 4 degrees C for 24 h before being frozen, was compared in a pilot metabolomic study of samples from 40 separate healthy volunteers. METHODS: Metabolic profiling with GC-TOF-MS was performed for serum and urine samples collected from 40 volunteers and stored at -80 degrees C or 4 degrees C for 24 h before being frozen at -80 degrees C. Subsequent Wilcoxon rank sum test and Principal Components Analysis (PCA) methods were used to assess whether differences in the metabolomes were detected between samples stored at 4 degrees C for 0 or 24 h. RESULTS: More than 700 unique metabolite peaks were detected, with over 200 metabolite peaks detected in any one sample. PCA and Wilcoxon rank sum tests of serum and urine data showed as a general observation that the variance associated with the replicate analysis per sample (analytical variance) was of the same magnitude as the variance observed between samples stored at 4 degrees C for 0 or 24 h. From a functional point of view the metabolomic composition of the majority of samples did not change in a statistically significant manner when stored under two different conditions. CONCLUSIONS: Based on this small pilot study, the UK Biobank sampling, transport and fractionation protocols are considered suitable to provide samples, which can produce scientifically robust and valid data in metabolomic studies. PMID- 18381391 TI - High-throughput 1H NMR-based metabolic analysis of human serum and urine for large-scale epidemiological studies: validation study. AB - BACKGROUND: Metabolic profiling of biofluid specimens is an established method for investigating disease states in clinical studies but is only recently being applied to large-scale human population studies. As part of protocol development for the UK Biobank study, a (1)H nuclear magnetic resonance (NMR)-based metabonomic analysis of specimen storage effects and analytical reproducibility was carried out using urine and serum specimens from 40 volunteers. METHODS: Aliquots of each specimen were stored for t = 0 and t = 24 h at 4 degrees C prior to freezing, and in the case of serum samples for a further 12 h (t = 36), to determine whether the storage times affected specimen composition and quality. A blinded split-specimen matching exercise was implemented to assign candidate spectral pairs stored for different times using multivariate statistical analysis of the NMR data. RESULTS: Using a chemometric strategy, split specimens at time t = 0 and t = 24 or 36 h after storage at 4 degrees C were easily paired and the split-specimen matching task was reduced to a workable size. (1)H NMR profiling established that the t = 24 h urine and serum groups showed no systematic metabolite changes, indicating biochemical stability. Some small differences in serum specimens stored for t = 36 h at 4 degrees C were detectable only by multivariate analysis, and were attributed to generalized alterations in proteins and protein fragments, and possibly trimethylamine-N-oxide. No other specific metabolite was implicated. CONCLUSIONS: For the purposes of NMR-based analysis, storage of urine and serum for up to t = 24 h at 4 degrees C does not detectably affect the metabolic profile and the methodology is robust. Future application of multivariate methods to data-rich studies should substantially enhance information recovery from epidemiological studies. PMID- 18381392 TI - EBV Immortalization of human B lymphocytes separated from small volumes of cryo preserved whole blood. AB - EBV immortalized B lymphocyte cell lines have been extensively used as a source of biological material for functional and molecular studies and represent a potentially limitless source of genomic DNA. Current technologies for EBV transformation are costly and use relatively large volumes of peripheral blood. Alternative methods were examined to determine whether smaller volumes of cryo preserved whole blood could be subsequently transformed and which could provide a more cost-effective strategy for large population-based studies such as UK Biobank. A successful method was established where viable B cells were positively selected from 0.5 ml cryo-preserved whole blood samples. These were EBV transformed in microtitre plates and subsequently expanded in culture. A pilot study within UK Biobank was performed, which confirmed its potential usefulness for this study. PMID- 18381393 TI - Design and implementation of a high-throughput biological sample processing facility using modern manufacturing principles. AB - BACKGROUND: UK Biobank is a prospective study that is collecting biological samples and health and lifestyle data from 500 000 volunteer participants over a 4-year period. These data will be used to facilitate biological and medical research. METHODS: Modern manufacturing principles were used to direct the development of the sample processing facility and automated systems. RESULTS: A fit for purpose facility comprising technology, systems, dedicated process, infrastructure and an appropriate staff structure has been implemented that will deliver and maintain a resource that will support the long-term goals of the UK Biobank study. CONCLUSIONS: Modern manufacturing principles are appropriate for use in the development of a high throughput biological sample processing facility. PMID- 18381394 TI - Designing an automated blood fractionation system. AB - BACKGROUND: UK Biobank will be collecting blood samples from a cohort of 500 000 volunteers and it is expected that the rate of collection will peak at approximately 3000 blood collection tubes per day. These samples need to be prepared for long-term storage. It is not considered practical to manually process this quantity of samples so an automated blood fractionation system is required. METHODS: Principles of industrial automation were applied to the blood fractionation process leading to the requirement of developing a vision system to identify the blood fractions within the blood collection tube so that the fractions can be accurately aspirated and dispensed into micro-tubes. A prototype was manufactured and tested on a range of human blood samples collected in different tube types. RESULTS: A specially designed vision system was capable of accurately measuring the position of the plasma meniscus, plasma/buffy coat interface and the red cells/buffy coat interface within a vacutainer. A rack of 24 vacutainers could be processed in <5 min. CONCLUSION: An automated, high throughput blood fractionation system offers a solution to the problem of processing human blood samples collected in vacutainers in a consistent manner and provides a means of ensuring data and sample integrity. PMID- 18381395 TI - Designing and implementing a large-scale automated -80 degrees C archive. AB - BACKGROUND: This article describes an automated facility for storing biological samples at -80 degrees C, designed to meet the needs of The UK Biobank. No store of this size has previously used liquid nitrogen as refrigerant, and so it was important to confirm the storage compartments could achieve and maintain uniform conditions with efficient use of the coolant. In addition, the store incorporates a novel system of drawers designed to allow robotic access whilst maintaining storage conditions. Experiments were undertaken to prove the performance of both these factors in maintaining the temperature of stored samples, both in steady state conditions and during sample access. METHODS: Sample tubes containing water were placed in key locations within a prototype storage compartment. Thermocouples inside these sample tubes were used to together with automated data loggers to accumulate continuous records of the temperature of the samples during the experiments. RESULTS: Results show that the design of refrigeration system and storage compartment, using LN(2) as refrigerant, enables efficient use of coolant and maintains uniform temperature over the entire compartment. The results also show that samples within the compartment remain at the intended storage temperature during automated retrieval operations, including the situation when an access drawer remains open indefinitely. CONCLUSIONS: These results confirm that the store design meets the requirements for the UK Biobank, and have given both the manufacturer (The Automation Partnership) and the UK Biobank the confidence to proceed with the implementation of a full sized automated store to hold 10 million samples. PMID- 18381396 TI - Design and implementation of a large-scale liquid nitrogen archive. AB - UK Biobank required a cryogenic facility to store, in vapour phase, a minimum of 5 000 000 1.2 ml capacity micro-tubes held in racks of 96 tubes below -180 degrees C for a 20 year period. The archive would be housed in a 900 m(2) industrial unit remote from the main laboratory area that would be generally unmanned and would require a complete cryogenic infrastructure. A commercial supplier was commissioned to provide a complete system including design, project management, supply and installation of all the cryogenic storage requirement including bulk tank, super insulated vacuum lines, Taylor Wharton LABS80K high capacity long-term vapour phase storage vessels with inventory control systems and all the local and remote software. The large-scale storage vessels were selected to give a high storage density, temperature stability and storage in vapour phase. PMID- 18381397 TI - Biorepositories--at the bleeding edge. PMID- 18381398 TI - The UK Biobank sample handling and storage protocol for the collection, processing and archiving of human blood and urine. AB - BACKGROUND: UK Biobank is a large prospective study in the UK to investigate the role of genetic factors, environmental exposures and lifestyle in the causes of major diseases of late and middle age. Extensive data and biological samples are being collected from 500,000 participants aged between 40 and 69 years. The biological samples that are collected and how they are processed and stored will have a major impact on the future scientific usefulness of the UK Biobank resource. AIMS: The aim of the UK Biobank sample handling and storage protocol is to specify methods for the collection and storage of participant samples that give maximum scientific return within the available budget. Processing or storage methods that, as far as can be predicted, will preclude current or future assays have been avoided. METHODS: The protocol was developed through a review of the literature on sample handling and processing, wide consultation within the academic community and peer review. Protocol development addressed which samples should be collected, how and when they should be processed and how the processed samples should be stored to ensure their long-term integrity. The recommended protocol was extensively tested in a series of validation studies. UK Biobank collects about 45 ml blood and 9 ml of urine with minimal local processing from each participant using the vacutainer system. A variety of preservatives, anti coagulants and clot accelerators is used appropriate to the expected end use of the samples. Collection of other material (hair, nails, saliva and faeces) was also considered but rejected for the full cohort. Blood and urine samples from participants are transported overnight by commercial courier to a central laboratory where they are processed and aliquots of urine, plasma, serum, white cells and red cells stored in ultra-low temperature archives. Aliquots of whole blood are also stored for potential future production of immortalized cell lines. A standard panel of haematology assays is completed on whole blood from all participants, since such assays need to be conducted on fresh samples (whereas other assays can be done on stored samples). By the end of the recruitment phase, 15 million sample aliquots will be stored in two geographically separate archives: 9.5 million in a -80 degrees C automated archive and 5.5 million in a manual liquid nitrogen archive at -180 degrees C. Because of the size of the study and the numbers of samples obtained from participants, the protocol stipulates a highly automated approach for the processing and storage of samples. Implementation of the processes, technology, systems and facilities has followed best practices used in manufacturing industry to reduce project risk and to build in quality and robustness. The data produced from sample collection, processing and storage are highly complex and are managed by a commercially available LIMS system fully integrated with the entire process. CONCLUSION: The sample handling and storage protocol adopted by UK Biobank provides quality assured and validated methods that are feasible within the available funding and reflect the size and aims of the project. Experience from recruiting and processing the first 40,000 participants to the study demonstrates that the adopted methods and technologies are fit-for-purpose and robust. PMID- 18381399 TI - Commentary: People's vulnerability to heat wave. PMID- 18381400 TI - Commentary: Calculations of EPIC proportions. PMID- 18381401 TI - FT-COMAR: fault tolerant three-dimensional structure reconstruction from protein contact maps. AB - Fault Tolerant Contact Map Reconstruction (FT-COMAR) is a heuristic algorithm for the reconstruction of the protein three-dimensional structure from (possibly) incomplete (i.e. containing unknown entries) and noisy contact maps. FT-COMAR runs within minutes, allowing its application to a large-scale number of predictions. AVAILABILITY: http://bioinformatics.cs.unibo.it/FT-COMAR PMID- 18381402 TI - Expected gene-order distances and model selection in bacteria. AB - MOTIVATION: The evolutionary distance inferred from gene-order comparisons of related bacteria is dependent on the model. Therefore, it is highly important to establish reliable assumptions before inferring its magnitude. RESULTS: We investigate the patterns of dotplots between species of bacteria with the purpose of model selection in gene-order problems. We find several categories of data which can be explained by carefully weighing the contributions of reversals, transpositions, symmetrical reversals, single gene transpositions and single gene reversals. We also derive method of moments distance estimates for some previously uncomputed cases, such as symmetrical reversals, single gene reversals and their combinations, as well as the single gene transpositions edit distance. PMID- 18381403 TI - Assigning functional linkages to proteins using phylogenetic profiles and continuous phenotypes. AB - MOTIVATION: A class of non-homology-based methods for protein function prediction relies on the assumption that genes linked to a phenotypic trait are preferentially conserved among organisms that share the trait. These methods typically compare pairs of binary strings, where one string encodes the phylogenetic distribution of a trait and the other of a protein. In this work, we extended the approach to automatically deal with continuous phenotypes. RESULTS: Rather than use a priori rules, which can be very subjective, to construct binary profiles from continuous phenotypes, we propose to systematically explore thresholds which can meaningfully separate the phenotype values. We illustrate our method by analyzing optimal growth temperatures, and demonstrate its usefulness by automatically retrieving genes which have been associated with thermophilic growth. We also apply the general approach, for the first time, to optimal growth pH, and make novel predictions. Finally, we show that our method can also be applied to other properties which may not be classically considered as phenotypes. Specifically, we studied correlations between genome size and the distribution of genes. PMID- 18381404 TI - Use of atypical antipsychotic drugs for schizophrenia in Maine Medicaid following a policy change. AB - More than one-third of Medicaid programs and Medicare Part D plans use prior authorization (PA) policies to control the use of atypical antipsychotics (AAs). We used Medicaid and Medicare claims data to investigate how Maine's PA policy affected AA use, treatment discontinuities, and spending among schizophrenia patients initiating AA therapy. Patients initiating AAs during Maine's policy experienced a 29 percent greater risk of treatment discontinuity than patients initiating AAs before the policy took effect; no change occurred in a comparison state. AA spending was slightly lower in both states. Observed increases in treatment discontinuities without cost savings suggest that AAs should be exempt from PA for patients with severe mental illnesses. PMID- 18381405 TI - Identification of p18 INK4c as a tumor suppressor gene in glioblastoma multiforme. AB - Genomic alterations leading to aberrant activation of cyclin/cyclin-dependent kinase (cdk) complexes drive the pathogenesis of many common human tumor types. In the case of glioblastoma multiforme (GBM), these alterations are most commonly due to homozygous deletion of p16(INK4a) and less commonly due to genomic amplifications of individual genes encoding cyclins or cdks. Here, we describe deletion of the p18(INK4c) cdk inhibitor as a novel genetic alteration driving the pathogenesis of GBM. Deletions of p18(INK4c) often occurred in tumors also harboring homozygous deletions of p16(INK4a). Expression of p18(INK4c) was completely absent in 43% of GBM primary tumors studied by immunohistochemistry. Lentiviral reconstitution of p18(INK4c) expression at physiologic levels in p18(INK4c)-deficient but not p18(INK4c)-proficient GBM cells led to senescence like G(1) cell cycle arrest. These studies identify p18(INK4c) as a GBM tumor suppressor gene, revealing an additional mechanism leading to aberrant activation of cyclin/cdk complexes in this terrible malignancy. PMID- 18381406 TI - Risk stratification in cancer predisposition syndromes: lessons learned from novel molecular developments in Li-Fraumeni syndrome. AB - Germ-line mutations in specific genes predispose family members to cancer. Prediction of the exact tumor type and timing of cancer initiation is fundamental to the development of management strategies for these individuals. Recent advances in our understanding of the general processes that control cancer initiation may enable us to tailor more precise risk stratification. This, in turn, will lead to more effective early detection strategies, which would result in more favorable clinical outcomes. In this review, we highlight the steps and methods used to reach this futuristic model. PMID- 18381407 TI - Cell cycle-dependent ciliogenesis and cancer. AB - In mammals, most cell types have primary cilia, protruding structures involved in sensing mechanical and chemical signals from the extracellular environment that act as major communication hubs for signaling controlling cell differentiation and polarity. The list of clinical disorders associated with ciliary dysfunction has expanded from polycystic kidney disease to include many others. Transformed cells commonly lack cilia, but whether this lack is cause or consequence of transformation is not well understood. Here we discuss work addressing recently identified actions of the cancer-promoting proteins Aurora A and HEF1/NEDD9/CAS-L at cilia. Together with older studies, this work suggests that loss of cilia in cancer may contribute to the insensitivity of cancer cells to environmental repressive signals, based in part on derangement of cell cycle checkpoints governed by cilia and centrosomes. PMID- 18381408 TI - Mcl-1: a gateway to TRAIL sensitization. AB - The proapoptotic cytokine tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is being evaluated presently as a selective anticancer agent, but its limited effects against cancer cell lines has raised some concerns about its ultimate clinical utility. Here, we review recent findings that cancer cell sensitivity to TRAIL is greatly increased when the Bcl-2 family protein Mcl-1 is down-regulated by the Raf/vascular endothelial growth factor kinase inhibitor sorafenib, a Food and Drug Administration-approved cancer drug. Using the TRAIL sorafenib combination as a tactic to more effectively kill cancer cells may provide an effective tool to attack a variety of human cancers that are largely presently untreatable. PMID- 18381409 TI - MUC4 mucin interacts with and stabilizes the HER2 oncoprotein in human pancreatic cancer cells. AB - MUC4, a high-molecular weight transmembrane glycoprotein, is overexpressed in pancreatic cancer and is implicated in its pathogenesis. It is a heterodimeric protein containing a large extracellular, heavily glycosylated subunit, MUC4alpha, and a transmembrane growth factor-like subunit, MUC4beta. In the present study, we have shown the interaction of human MUC4 with the receptor tyrosine kinase HER2 in pancreatic adenocarcinoma cells by reciprocal coimmunoprecipitation and cocapping studies. MUC4 colocalized with HER2 at the cell surface and in the cytoplasm. Silencing of MUC4 by transient or stable expression of MUC4-targeted short-interfering RNA led to the down-regulation of HER2 with a concomitant decrease in its phosphorylated form (pY(1248)-HER2). Further analyses revealed that the MUC4-knockdown-mediated decrease in HER2 expression occurred due to the drop in the stability of the receptor. In MUC4 knockdown pancreatic cancer cells, we also observed a reduced phosphorylation of the focal adhesion kinase and p42/44 mitogen-activated protein kinase, which are downstream effector proteins in HER2 signaling. Our findings add a new dimension to MUC4 function as a modulator of cell signaling and provide mechanistic evidence for its role in pancreatic cancer progression. PMID- 18381410 TI - Enhancing poxvirus oncolytic effects through increased spread and immune evasion. AB - The antitumoral effects of oncolytic viruses have generally been limited by inefficient spread of the viruses within infected tumors and by inefficient systemic delivery, particularly in preimmunized hosts. Tumor-selective poxviruses have biological characteristics that may overcome these limitations. Nevertheless, physical barriers within the tumor microenvironment, including the extracellular matrix, can still limit intratumoral spread, and neutralizing antibodies can impede systemic delivery. To counter these limitations, we sought to take advantage of a naturally occurring poxvirus form known as extracellular enveloped virus (EEV). The EEV is shrouded by a host cell-derived lipid bilayer containing anticomplement proteins and is typically released from infected cells early during the infection cycle. Therefore, the EEV form evolved for rapid systemic spread within the host and for evasion of immune-mediated clearance. We compared the oncolytic potential of low versus high EEV-producing strains of vaccinia. EEV-enhanced vaccinia strains displayed improved spread within tumors after systemic delivery, resulting in significantly improved antitumor effects. The EEV-enhanced strains also displayed a greater ability to spread between injected and noninjected distant tumors through the blood and, importantly, displayed reduced clearance by neutralizing antibody. Safety was unaffected. The incorporation of EEV-enhancing mutations into next generation oncolytic vaccinia strains may improve the potency of these viruses without sacrificing safety. PMID- 18381411 TI - Bisphenol A induces a profile of tumor aggressiveness in high-risk cells from breast cancer patients. AB - Breast cancer outcome is highly variable. Whether inadvertent exposure to environmental xenobiotics evokes a biological response promoting cancer aggressiveness and a higher probability of tumor recurrence remains unknown. To determine specific molecular alterations which arise in high-risk breast tissue in the presence of the ubiquitous xenoestrogen, bisphenol A (BPA), we used nonmalignant random periareolar fine-needle aspirates in a novel functional assay. Early events induced by BPA in epithelial-stromal cocultures derived from the contralateral tissue of patients with breast cancer included gene expression patterns which facilitate apoptosis evasion, endurance of microenvironmental stress, and cell cycle deregulation without a detectable increase in cell numbers. This BPA response profile was significantly associated with breast tumors characterized by high histologic grade (P < 0.001) and large tumor size (P = 0.002), resulting in decreased recurrence-free patient survival (P < 0.001). Our assays show a biological "fingerprint" of probable prior exposure to endocrine-disrupting agents, and suggest a scenario in which their presence in the microenvironmental milieu of high-risk breast tissue could play a deterministic role in establishing and maintaining tumor aggressiveness and poor patient outcome. PMID- 18381412 TI - Mutation frequencies and spectra in DNA polymerase eta-deficient mice. AB - The low-fidelity polymerase eta (poleta) is required for bypass of UV-induced pyrimidine dimers inserting adenine nucleotides opposite these lesions. Mutations in the poleta gene are responsible for the genetic defect in xeroderma pigmentosum variant patients. To study if the lack of poleta significantly elevates spontaneous mutation frequency in various organs and tissues of the mouse, we crossed poleta-deficient mice with transgenic mice harboring a chromosomally integrated lacZ-plasmid reporter construct. In cultured embryonic fibroblasts from the lacZ-poleta(-/-) mice, 2.5 J/m(2) UV irradiation induced approximately 5-fold more mutations than in cells from lacZ control mice, in which an approximately 3-fold increase in mutation frequency was found compared with the normal level. Whereas untreated cells harbored mainly 1-bp deletions, UV induced both transitions and transversions, with the latter type more highly represented in the poleta-null cells than in the controls. No difference in mutation induction between the poleta-null cells and the wild-type cells was observed after treatment with N-ethyl-N-nitrosourea. Having shown the validity of the lacZ model to accurately identify poleta-associated mutagenesis, we then determined the mutant frequency at the lacZ locus in liver, spleen, and small intestine of 12-month-old animals. No differences were found between poleta-null, heterozygous, or littermate control mice. We conclude that the poleta defect is specific for UV damage and has no effect on in vivo mutagenesis in mice. PMID- 18381413 TI - Pancreatic stellate cells: partners in crime with pancreatic cancer cells. AB - Pancreatic stellate cells (PSC) produce the stromal reaction in pancreatic cancer, but their role in cancer progression is not fully elucidated. We examined the influence of PSCs on pancreatic cancer growth using (a) an orthotopic model of pancreatic cancer and (b) cultured human PSCs (hPSC) and human pancreatic cancer cell lines MiaPaCa-2 and Panc-1. Athymic mice received an intrapancreatic injection of saline, hPSCs, MiaPaCa-2 cells, or hPSCs + MiaPaCa-2. After 7 weeks, tumor size, metastases, and tumor histology were assessed. In vitro studies assessed the effect of cancer cell secretions on PSC migration and the effect of hPSC secretions on cancer cell proliferation, apoptosis, and migration. Possible mediators of the effects of hPSC secretions on cancer cell proliferation were examined using neutralizing antibodies. Compared with mice receiving MiaPaCa-2 cells alone, mice injected with hPSCs + MiaPaCa-2 exhibited (a) increased tumor size and regional and distant metastasis, (b) fibrotic bands (desmoplasia) containing activated PSCs within tumors, and (c) increased tumor cell numbers. In vitro studies showed that, in the presence of pancreatic cancer cells, PSC migration was significantly increased. Furthermore, hPSC secretions induced the proliferation and migration, but inhibited the apoptosis, of MiaPaCa-2 and Panc-1 cells. The proliferative effect of hPSC secretions on pancreatic cancer cells was inhibited in the presence of neutralizing antibody to platelet-derived growth factor. Our studies indicate a significant interaction between pancreatic cancer cells and stromal cells (PSCs) and imply that pancreatic cancer cells recruit stromal cells to establish an environment that promotes cancer progression. PMID- 18381414 TI - Exploration of tumor-suppressive microRNAs silenced by DNA hypermethylation in oral cancer. AB - In the last few years, microRNAs (miRNA) have started a revolution in molecular biology and emerged as key players in the carcinogenesis. They have been identified in various tumor types, showing that different sets of miRNAs are usually deregulated in different cancers. To identify the miRNA signature that was specific for oral squamous cell carcinoma (OSCC), we first examined expression profiles of 148 miRNAs in a panel of 18 OSCC cell lines and the immortalized oral keratinocyte line RT7 as a control. Compared with RT7, the expression of 54 miRNAs (36.5%) was frequently down-regulated in OSCC lines (<0.5 fold expression, >or=66.7% of 18 lines). Among these 54 miRNAs, we further analyzed four of these miRNAs (i.e., miR-34b, miR-137, miR-193a, and miR-203), located around CpG islands, to identify tumor-suppressive miRNAs silenced through aberrant DNA methylation. The expression of those four genes was restored by treatment with 5-aza-2'-deoxycytidine in OSCC cells lacking their expression. In addition, expression levels of the four miRNAs were inversely correlated with their DNA methylation status in the OSCC lines. In primary tumors of OSCC with paired normal oral mucosa, down-regulation of miRNA expression through tumor specific hypermethylation was more frequently observed for miR-137 and miR-193a than for miR-34b and miR-203. Moreover, the ectopic transfection of miR-137 or miR-193a into OSCC lines lacking their expressions significantly reduced cell growth, with down-regulation of the translation of cyclin-dependent kinase 6 or E2F transcription factor 6, respectively. Taken together, our results clearly show that miR-137 and miR-193a are tumor suppressor miRNAs epigenetically silenced during oral carcinogenesis. PMID- 18381415 TI - Epidermal growth factor receptor gene amplification is acquired in association with tumor progression of EGFR-mutated lung cancer. AB - Both mutation and amplification of epidermal growth factor receptor (EGFR) in lung cancers have been reported in association with clinical responses to tyrosine kinase inhibitors. We have reported evidence implicating mutation specifically in the "terminal respiratory unit" type of adenocarcinoma, which is characterized by expression of thyroid transcription factor 1, a lineage marker of peripheral airway cells. However, little is known about the role of gene amplification in the molecular progression of lung adenocarcinoma. In this study, we examined the topographical distribution of amplification in three microdissected portions each of 48 individual lung cancers with confirmed mutations. Relative copy number of the gene was analyzed using Taq Man-based gene dosage analysis and fluorescent in situ hybridization technique. Gene amplification was found in 11 lung cancers. Strikingly, nine of the cancers showed heterogeneous distribution, and amplification was associated with higher histologic grade or invasive growth. Because it was likely that the high-grade lesions were the origin for metastases, metastatic lymph nodes corresponding to five tumors with heterogeneous distribution were analyzed. Unexpectedly, amplification status of the metastatic sites was not always associated with gene amplification of the primary tumors, suggesting that selection of the metastatic clone may be defined by other factors. We also examined 17 precursor lesions and 21 in situ lung adenocarcinomas, and found that only one in situ carcinoma harbored gene amplification. Taken together, our results show that mutation occurs early in the development of lung adenocarcinoma, and that amplification may be acquired in association with tumor progression. PMID- 18381416 TI - Methylation silencing of transforming growth factor-beta receptor type II in rat prostate cancers. AB - To identify methylation-silenced genes in prostate cancers, a microarray analysis for genes up-regulated by treatment with a demethylating agent, 5-aza-2' deoxycytidine, was performed using three rat prostate cancer cell lines. Eight genes (Aebp1, Dysf, Gas6, LOC361288, Nnat, Ocm, RGD1308119, and Tgfbr2) were re expressed at 16-fold or more, and their promoter CpG islands were shown to be densely methylated in the cancer cell lines. From the eight genes, Tgfbr2, a key mediator of transforming growth factor-beta (TGF-beta) signaling that has been strongly implicated in human and rat prostate carcinogenesis, was selected, and its silencing in primary samples was analyzed further. Tgfbr2 was methylated and markedly down-regulated in three of seven 3,2'-dimethyl-4-aminobiphenyl-induced invasive adenocarcinomas in the dorsolateral lobe of the rat prostate. In humans, marked down-regulation of TGFBR2 protein was observed in 12 of 20 high-grade prostatic intraepithelial neoplasia and 36 of 60 prostate cancers. DNA methylation of the human TGFBR2 promoter CpG islands repressed transcription, if present, but neither methylation nor mutation were detected in 27 human prostate cancers analyzed. Methylation silencing of rat Tgfbr2 was associated with histone H3 lysine 9 trimethylation, whereas decreased expression of human TGFBR2 was mainly due to decreased transcription activity, sometimes in concert with histone deacetylation and H3 lysine 27 trimethylation. The identification of methylation silencing of Tgfbr2 in rat prostate cancers, in accordance with TGFBR2 down regulation in human prostate cancers, will enable us to analyze how aberrant methylation is induced in vivo and identify factors that promote and suppress the induction of aberrant methylation. PMID- 18381417 TI - Phosphatase and tensin homologue deleted on chromosome 10 deficiency accelerates tumor induction in a mouse model of ErbB-2 mammary tumorigenesis. AB - Loss of the tumor suppressor phosphatase and tensin homologue deleted on chromosome 10 (PTEN) and amplification or elevated expression of ErbB-2 are both involved in human breast cancer. To directly test the importance of these genetic events in mammary tumorigenesis, we have assessed whether mammary-specific disruption of PTEN could cooperate with activation of ErbB-2. Transgenic mice expressing ErbB-2 under the transcriptional control of its endogenous promoter (ErbB-2(KI)) were interbred with mice carrying conditional PTEN alleles and an MMTV/Cre transgene. Loss of one or both PTEN alleles resulted in a dramatic acceleration of mammary tumor onset and an increased occurrence of lung metastases in the ErbB-2(KI) strain. Tumor progression in PTEN-deficient/ErbB 2(KI) strains was associated with elevated ErbB-2 protein levels, which were not due to ErbB-2 amplification or to a dramatic increase in ErbB-2 transcripts. Moreover, the PTEN-deficient/ErbB-2(KI)-derived mouse mammary tumors display striking morphologic heterogeneity in comparison with the homogeneous pathology of the ErbB-2(KI) parental strain. Therefore, inactivation of PTEN would not only have a dramatic effect on ErbB-2-induced mammary tumorigenesis but would also lead to the formation of mammary tumors that, in part, display pathologic and molecular features associated with the basal-like subtype of primary human breast cancer. PMID- 18381418 TI - Activator protein-1 transcription factors are associated with progression and recurrence of prostate cancer. AB - To identify biomarkers that discriminate the aggressive forms of prostate cancer, we performed gene expression profiling of prostate tumors using a genetically engineered mouse model that recapitulates the stages of human prostate cancer, namely Nkx3.1; Pten mutant mice. We observed a significant deregulation of the epidermal growth factor and mitogen-activated protein kinase (MAPK) signaling pathways, as well as their major downstream effectors--the activator protein-1 transcription factors c-Fos and c-Jun. Forced expression of c-Fos and c-Jun in prostate cancer cells promotes tumorigenicity and results in activation of extracellular signal-regulated kinase (Erk) MAPK signaling. In human prostate cancer, up-regulation of c-Fos and c-Jun proteins occurs in advanced disease and is correlated with Erk MAPK pathway activation, whereas high levels of c-Jun expression are associated with disease recurrence. Our analyses reveal a hitherto unappreciated role for AP-1 transcription factors in prostate cancer progression and identify c-Jun as a marker of high-risk prostate cancer. This study provides a striking example of how accurate mouse models can provide insights on molecular processes involved in progression and recurrence of human cancer. PMID- 18381419 TI - Origins and prevalence of the American Founder Mutation of MSH2. AB - Large germline deletions within the mismatch repair gene MSH2 account for a significant proportion (up to 20%) of all deleterious mutations of this gene which are associated with Lynch syndrome. An exons 1 to 6 deletion of MSH2, originally reported in nine families, has been associated with a founding event within the United States, which genealogic studies had previously dated to 1727, and the number of present day carriers was estimated to be 18,981. Here, we report the development of a robust multiplex PCR which has assisted in the detection of 32 new families who carry the MSH2 American Founder Mutation (AFM). By offering testing to family members, 126 carriers of the AFM have been identified. Extensive genealogic studies have connected 27 of the 41 AFM families into seven extended pedigrees. These extended families have been traced back to around the 18th century without any evidence of further convergence between them. Characterization of the genomic sequence flanking the deletion and the identification of a common disease haplotype of between 0.6 and 2.3 Mb in all probands provides evidence for a common ancestor between these extended families. The DMLE+2.2 software predicts an age of approximately 500 years (95% confidence interval, 425-625) for this mutation. Taken together, these data are suggestive of an earlier founding event than was first thought, which likely occurred in a European or a Native American population. The consequences of this finding would be that the AFM is significantly more frequent in the United States than was previously predicted. PMID- 18381420 TI - Frequency of the CHEK2 1100delC mutation among women with breast cancer: an international study. AB - A founder allele in the CHEK2 gene (1100delC) has been associated with an elevated risk of breast cancer. This allele is responsible for the majority of CHEK2-associated breast cancers in women from northern European countries; however, within Europe, it seems to be rare in countries that are close to the Mediterranean. The frequency of the 1100delC allele has not been measured in non White populations. We measured the frequency of the CHEK2 founder allele in 3,882 breast cancer patients and 8,609 controls from various countries. The allele was not seen among Asian patients (from Pakistan or the Philippines) and was present in 1 of 155 cases from Brazil. Among White women, the allele was present in 1.5% of 825 familial cases of breast cancer and in 0.7% of 1,106 patients with nonfamilial breast cancer. The allele was equally frequent in Jewish and non Jewish patients. We estimate that the CHEK2 1100delC allele is associated with an odds ratio of 2.6 for breast cancer, which corresponds to a lifetime risk of approximately 24% in Ontario. PMID- 18381421 TI - Hypoxia down-regulates CCAAT/enhancer binding protein-alpha expression in breast cancer cells. AB - The transcription factor CCAAT/enhancer binding protein-alpha (C/EBP alpha) is involved in the control of cell differentiation and proliferation, and has been suggested to act as a tumor suppressor in several cancers. By using microarray analysis, we have previously shown that hypoxia and estrogen down-regulate C/EBP alpha mRNA in T-47D breast cancer cells. Here, we have examined the mechanism by which the down-regulation by hypoxia takes place. Using the specific RNA polymerase II inhibitor 5,6-dichlorobenzimidazole-1-beta-D-ribofuranoside, the mRNA stability was analyzed under normoxia or hypoxia by quantitative reverse transcription-PCR. Hypoxia reduced the half-life of C/EBP alpha mRNA by approximately 30%. C/EBP alpha gene promoter studies indicated that hypoxia also repressed the transcription of the gene and identified a hypoxia-responsive element (-522; -527 bp), which binds to hypoxia-inducible factor (HIF)-1 alpha, as essential for down-regulation of C/EBP alpha transcription in hypoxia. Immunocytochemical analysis showed that C/EBP alpha was localized in the nucleus at 21% O(2), but was mostly cytoplasmic under 1% O(2). Knockdown of HIF-1 alpha by RNAi restored C/EBP alpha to normal levels under hypoxic conditions. Immunohistochemical studies of 10 tumor samples did not show any colocalization of C/EBP alpha and glucose transporter 1 (used as a marker for hypoxia). Taken together, these results show that hypoxia down-regulates C/EBP alpha expression in breast cancer cells by several mechanisms, including transcriptional and posttranscriptional effects. The down-regulation of C/EBP alpha in hypoxia is mediated by HIF-1. PMID- 18381422 TI - c-Jun NH2-terminal kinase activating kinase 1/mitogen-activated protein kinase kinase 4-mediated inhibition of SKOV3ip.1 ovarian cancer metastasis involves growth arrest and p21 up-regulation. AB - In many patients without clinical metastases, cancer cells have already escaped from the primary tumor and entered a distant organ. A long-standing question in metastasis research is why some disseminated cancer cells fail to complete steps of metastatic colonization for extended periods of time. Our laboratory identified c-Jun NH(2)-terminal kinase activating kinase 1/mitogen-activated protein kinase kinase 4 (JNKK1/MKK4) as a metastasis suppressor protein in a mouse xenograft model of experimental i.p. ovarian cancer metastasis. In this model, expression of JNKK1/MKK4 via activation of p38 delays formation of >or=1 mm implants and prolongs animal survival. Here, we elucidate the time course of this delay as well as the biological mechanisms underpinning it. Using the Gompertz function to model the net accumulation of experimental omental metastases, we show that MKK4-expressing implants arise, on average, 30 days later than controls. Quantitative real-time PCR shows that MKK4 expression does not have a substantial effect on the number of cancer cells initially adhering to the omentum, and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling analysis shows that there is no increase in apoptosis in these cells. Instead, immunohistochemical quantitation of cell cycle proteins reveals that MKK4-expressing cells fail to proliferate once they reach the omentum and up regulate p21, a cell cycle inhibitor. Consistent with the time course data, in vitro kinase assays and in vivo passaging of cell lines derived from macroscopic metastases show that the eventual outgrowth of MKK4-expressing cells is not due to a discrete selection event. Rather, the population of MKK4-expressing cells eventually uniformly adapts to the consequences of up-regulated MKK4 signaling. PMID- 18381423 TI - EWS-FLI-1 expression triggers a Ewing's sarcoma initiation program in primary human mesenchymal stem cells. AB - Ewing's sarcoma family tumors (ESFT) express the EWS-FLI-1 fusion gene generated by the chromosomal translocation t(11;22)(q24;q12). Expression of the EWS-FLI-1 fusion protein in a permissive cellular environment is believed to play a key role in ESFT pathogenesis. However, EWS-FLI-1 induces growth arrest or apoptosis in differentiated primary cells, and the identity of permissive primary human cells that can support its expression and function has until now remained elusive. Here we show that expression of EWS-FLI-1 in human mesenchymal stem cells (hMSC) is not only stably maintained without inhibiting proliferation but also induces a gene expression profile bearing striking similarity to that of ESFT, including genes that are among the highest ESFT discriminators. Expression of EWS-FLI-1 in hMSCs may recapitulate the initial steps of Ewing's sarcoma development, allowing identification of genes that play an important role early in its pathogenesis. Among relevant candidate transcripts induced by EWS-FLI-1 in hMSCs, we found the polycomb group gene EZH2, which we show to play a critical role in Ewing's sarcoma growth. These observations are consistent with our recent findings using mouse mesenchymal progenitor cells and provide compelling evidence that hMSCs are candidate cells of origin of ESFT. PMID- 18381424 TI - DNA damage signaling in hematopoietic cells: a role for Mre11 complex repair of topoisomerase lesions. AB - The Mre11 complex promotes DNA double-strand break repair and regulates DNA damage signaling via activation of the ataxia-telangiectasia mutated (ATM) kinase. The hypermorphic Rad50(S) allele encodes a variant of Rad50, a member of the Mre11 complex. Cells expressing Rad50(S) experience constitutive ATM activation, which leads to precipitous apoptotic attrition in hematopoietic cells. In this study, we show that ATM activation by the Rad50S-containing Mre11 complex enhances the proliferation of LSK cells, a population consisting of hematopoietic stem cells and multipotent progenitor cells. In Rad50(S/S) mice, enhanced LSK proliferation triggers apoptotic attrition. This phenotype is mitigated when Rad50(S/S) is combined with mutations that alter either LSK cell quiescence (myeloid elf-1-like factor/ELF4-deficient mice) or hematopoietic differentiation (p21- and p27-deficient mice), indicating that the LSK population is a primary target of Rad50(S) pathology. We show that cells from Rad50(S/S) mice are hypersensitive to camptothecin, a topoisomerase I inhibitor that causes DNA damage primarily during DNA replication. On this basis, we propose that apoptotic attrition of Rad50(S/S) hematopoietic cells results from enhanced proliferation in the context of topoisomerase-associated DNA damage. Impairment of apoptosis in Rad50(S/S) mice promotes hematopoietic malignancy, suggesting that primitive hematopoietic cells serve as a reservoir of potentially oncogenic lesions in Rad50(S/S) mice. These data provide compelling evidence that the Mre11 complex plays a role in the metabolism of topoisomerase lesions in mammals, and further suggest that such lesions can accumulate in primitive hematopoietic cells and confer significant oncogenic potential. PMID- 18381425 TI - Attenuation of junctional adhesion molecule-A is a contributing factor for breast cancer cell invasion. AB - The metastatic potential of cancer cells is directly attributed to their ability to invade through the extracellular matrix. The mechanisms regulating this cellular invasiveness are poorly understood. Here, we show that junctional adhesion molecule A (JAM-A), a tight junction protein, is a key negative regulator of cell migration and invasion. JAM-A is robustly expressed in normal human mammary epithelium, and its expression is down-regulated in metastatic breast cancer tumors. In breast cancer cell lines, an inverse relationship between JAM-A expression and the ability of these cells to migrate on a collagen matrix was observed, which correlates with the known ability of these cells to metastasize. The T47D and MCF-7 cells, which migrate least, are found to express high levels of JAM-A, whereas the more migratory MDA-MB-468 cells have lower levels of JAM-A on the cell surface. MDA-MB-231 cells, which are highly migratory, express the least amount of JAM-A. Overexpression of JAM-A in MDA-MB 231 cells inhibited both migration and invasion through collagen gels. Furthermore, knockdown of JAM-A using short interfering RNAs enhanced the invasiveness of MDA-MB-231 cells as well as T47D cells. The ability of JAM-A to attenuate cell invasion correlated with the formation of increased numbers of focal adhesions and the formation of functional tight junctions. These results show for the first time that an immunoglobulin superfamily cell adhesion protein expressed at tight junctions could serve as a key negative regulator of breast cancer cell invasion and possibly metastasis. Furthermore, loss of JAM-A could be used as a biomarker for aggressive breast cancer. PMID- 18381426 TI - Six1 overexpression in mammary cells induces genomic instability and is sufficient for malignant transformation. AB - Homeoproteins are transcription factors that act as master regulators of development and are frequently dysregulated in cancers. During embryogenesis, the Six1 homeoprotein is essential for the expansion of precursor cell populations that give rise to muscle and kidney, among other organs. Six1 overexpression is observed in numerous cancers, resulting in increased proliferation, survival, and metastasis. Here, we investigate whether Six1 can play a causal role in mammary tumor initiation. We show that Six1 overexpression in MCF12A mammary epithelial cells promotes multiple properties associated with malignant transformation, including increased proliferation, genomic instability, and anchorage-independent growth. We further show that this transformation is dependent on up-regulation of its transcriptional target, cyclin A1, which is normally expressed in the embryonic mammary gland but dramatically reduced in the adult gland. Six1 transformed MCF12A cells are tumorigenic in nude mice, forming aggressive tumors that are locally invasive and exhibit peritumoral lymphovascular invasion. In human breast carcinomas, expression of Six1 and cyclin A1 mRNA correlate strongly with each other (P < 0.0001), and expression of Six1 and cyclin A1 each correlate with Ki67, a marker of proliferation (P < 0.0001 and P = 0.014, respectively). Together, our data indicate that Six1 overexpression is sufficient for malignant transformation of immortalized, nontumorigenic mammary epithelial cells, and suggest that the mechanism of this transformation involves inappropriate reexpression of cyclin A1 in the adult mammary gland. PMID- 18381427 TI - Elevated ornithine decarboxylase levels activate ataxia telangiectasia mutated DNA damage signaling in normal keratinocytes. AB - We examined the effect of increased expression of ornithine decarboxylase (ODC), a key rate-limiting enzyme in polyamine biosynthesis, on cell survival in primary cultures of keratinocytes isolated from the skin of K6/ODC transgenic mice (Ker/ODC) and their normal littermates (Ker/Norm). Although elevated levels of ODC and polyamines stimulate proliferation of keratinocytes, Ker/ODC undergo apoptotic cell death within days of primary culture unlike Ker/Norm that continue to proliferate. Phosphorylation of ataxia telangiectasia mutated (ATM) and its substrate p53 are significantly induced both in Ker/ODC and in K6/ODC transgenic skin. Chromatin immunoprecipitation analyses show that the increased level of p53 in Ker/ODC is accompanied by increased recruitment of p53 to the Bax proximal promoter. ATM activation is polyamine dependent because alpha difluoromethylornithine, a specific inhibitor of ODC activity, blocks its phosphorylation. Ker/ODC also displays increased generation of H(2)O(2), acrolein lysine conjugates, and protein oxidation products as well as polyamine-dependent DNA damage, as measured by the comet assay and the expression of the phosphorylated form of the histone variant gamma H2AX. Both reactive oxygen species generation and apoptotic cell death of Ker/ODC may, at least in part, be due to induction of a polyamine catabolic pathway that generates both H(2)O(2) and cytotoxic aldehydes, because spermine oxidase (SMO) levels are induced in Ker/ODC. In addition, treatment with MDL 72,527, an inhibitor of SMO, blocks the production of H(2)O(2) and increases the survival of Ker/ODC. These results show a novel activation of the ATM-DNA damage signaling pathway in response to increased ODC activity in nontumorigenic keratinocytes. PMID- 18381428 TI - Lkb1 deficiency causes prostate neoplasia in the mouse. AB - Mutation of LKB1 is the key molecular event underlying Peutz-Jeghers syndrome, a dominantly inherited condition characterized by a predisposition to a range of malignancies, including those of the reproductive system. We report here the use of a Cre-LoxP strategy to directly address the role of Lkb1 in prostate neoplasia. Recombination of a LoxP-flanked Lkb1 allele within all four murine prostate lobes was mediated by spontaneous activation of a p450 CYP1A1-driven Cre recombinase transgene (termed AhCre). Homozygous mutation of Lkb1 in males expressing AhCre reduced longevity, with 100% manifesting atypical hyperplasia and 83% developing prostate intraepithelial neoplasia (PIN) of the anterior prostate within 2 to 4 months. We also observed focal hyperplasia of the dorsolateral and ventral lobes (61% and 56% incidence, respectively), bulbourethral gland cysts associated with atypical hyperplasia (100% incidence), hyperplasia of the urethra (39% incidence), and seminal vesicle squamous metaplasia (11% incidence). PIN foci overexpressed nuclear beta-catenin, p-Gsk3 beta, and downstream Wnt targets. Immunohistochemical analysis of foci also showed a reduction in Pten activation and up-regulation of both p-PDK1 (an AMPK kinase) and phosphorylated Akt. Our data are therefore consistent with deregulation of Wnt and phosphoinositide 3-kinase/Akt signaling cascades after loss of Lkb1 function. For the first time, this model establishes a link between the tumor suppressor Lkb1 and prostate neoplasia, highlighting a tumor suppressive role within the mouse and raising the possibility of a similar association in the human. PMID- 18381429 TI - Mitochondrial function and nuclear factor-kappaB-mediated signaling in radiation induced bystander effects. AB - Although radiation-induced bystander effects have been well described over the past decade, the mechanisms of the signaling processes involved in the bystander phenomenon remain unclear. In the present study, using the Columbia University charged particle microbeam, we found that mitochondrial DNA-depleted human skin fibroblasts (rho(o)) showed a higher bystander mutagenic response in confluent monolayers when a fraction of the same population were irradiated with lethal doses compared with their parental mitochondrial-functional cells (rho(+)). However, using mixed cultures of rho(o) and rho(+) cells and targeting only one population of cells with a lethal dose of alpha-particles, a decreased bystander mutagenesis was uniformly found in nonirradiated bystander cells of both cell types, indicating that signals from one cell type can modulate expression of bystander response in another cell type. In addition, we found that Bay 11-7082, a pharmacologic inhibitor of nuclear factor-kappaB (NF-kappaB) activation, and 2 (4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, a scavenger of nitric oxide (NO), significantly decreased the mutation frequency in both bystander rho(o) and rho(+) cells. Furthermore, we found that NF-kappaB activity and its dependent proteins, cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS), were lower in bystander rho(o) cells when compared with their rho(+) counterparts. Our results indicated that mitochondria play an important role in the regulation of radiation-induced bystander effects and that mitochondria dependent NF-kappaB/iNOS/NO and NF-kappaB/COX-2/prostaglandin E2 signaling pathways are important to the process. PMID- 18381430 TI - Gli activity correlates with tumor grade in platelet-derived growth factor induced gliomas. AB - Gli signaling is critical for central nervous system development and is implicated in tumorigenesis. To monitor Gli signaling in gliomas in vivo, we created platelet-derived growth factor-induced gliomas in a Gli-luciferase reporter mouse. We find that Gli activation is found in gliomas and correlates with grade. In addition, we find that sonic hedgehog (SHH) is expressed in these tumors and also correlates with grade. We identify microvascular proliferation and pseudopalisades, elements that define high-grade gliomas as SHH-producing microenvironments. We describe two populations of SHH-producing stromal cells that reside in perivascular niche (PVN), namely low-cycling astrocytes and endothelial cells. Using the Ptc-LacZ knock-in mouse as a second Gli responsive reporter, we show beta-galactosidase activity in the PVN and in some tumors diffusely throughout the tumor. Lastly, we observe that SHH is similarly expressed in human gliomas and note that an intact tumor microenvironment or neurosphere conditions in vitro are required for Gli activity. PMID- 18381431 TI - Regulation of Id1 expression by SRC: implications for targeting of the bone morphogenetic protein pathway in cancer. AB - Deregulated activation of the Src tyrosine kinase and heightened Id1 expression are independent mediators of aggressive tumor biology. The present report implicates Src signaling as a critical regulator of Id1 gene expression. Microarray analyses showed that Id family genes were among the most highly down regulated by incubation of A549 lung carcinoma cells with the small-molecule Src inhibitor AZD0530. Id1 transcript and protein levels were potently reduced in a dose-dependent manner concomitantly with the reduction of activated Src levels. These effects were conserved across a panel of lung, breast, prostate, and colon cancer cell lines and confirmed by the ability of PP2, Src siRNA, and Src blocking peptides to suppress Id1 expression. PP2, AZD0530, and dominant-negative Src abrogated Id1 promoter activity, which was induced by constitutively active Src. The Src-responsive region of the Id1 promoter was mapped to a region 1,199 to 1,360 bps upstream of the translation start site and contained a Smad-binding element. Src was also required for bone morphogenetic protein-2 (BMP-2)-induced Id1 expression and promoter activity, was moderately activated by BMP-2, and complexed with Smad1/5. Conversely, Src inhibitors blocked Smad1/5 nuclear translocation and binding to the Src-responsive region of the Id1 promoter. Consistent with a role for Src and Id1 in cancer cell invasion, Src inhibitors and Id1 siRNA decreased cancer cell invasion, which was increased by Id1 overexpression. Taken together, these results reveal that Src positively interacts with the BMP-Smad-Id pathway and provide new ways for targeted inhibition of Id1. PMID- 18381432 TI - The role of amphiregulin in exemestane-resistant breast cancer cells: evidence of an autocrine loop. AB - Exemestane-resistant breast cancer cell lines (i.e., ExeR), derived from MCF-7 cells expressing a high level of aromatase (MCF-7aro), were generated in our laboratory. The epidermal growth factor (EGF)-like protein amphiregulin (AREG) was highly expressed in ExeR cells based on cDNA microarray analysis. The high levels of AREG mRNA in ExeR cell lines were confirmed by real-time reverse transcription-PCR. The high levels of AREG protein in ExeR cell lysates and culture media were confirmed by Western blot analysis and ELISA, respectively. Furthermore, our Western blot analysis showed that whereas no AREG was detected in the DMSO control, overnight treatment of parental MCF-7aro cells with 1 micromol/L exemestane strongly induced the expression of AREG. This induction was totally blocked by 100 nmol/L of pure antiestrogen ICI 182,780, implying estrogen receptor (ER) dependence of exemestane-induced AREG expression. MCF-7aro cells were not able to proliferate in hormone-free medium, but were able to proliferate in conditioned medium from ExeR cells, similar to the treatment of recombinant human AREG. Small interference RNA targeting AREG inhibited ExeR proliferation, confirming that AREG is truly functioning as a growth factor of ExeR cells. The specific inhibitors to ER (ICI 182,780), EGF receptor (EGFR; AG1478), and mitogen activated protein kinase (MAPK; U0126) all showed dose-dependent suppression of the proliferation of ExeR cells, indicating the involvement of the ER, EGFR, and MAPK pathways. Based on these findings, we propose a possible mechanism that underlies exemestane resistance: exemestane induces AREG in an ER-dependent manner. AREG then activates the EGFR pathway and leads to the activation of the MAPK pathway that drives cell proliferation. PMID- 18381433 TI - Down-regulation of mammalian sterile 20-like kinase 1 by heat shock protein 70 mediates cisplatin resistance in prostate cancer cells. AB - Mammalian sterile 20-like kinase 1 (Mst1) is an ubiquitously expressed serine/threonine kinase, and its activation results in cell apoptosis. Recent studies suggest that Mst1 may function as a tumor suppressor. Here, we reported that heat shock protein 70 (Hsp70), which is thought to protect cells against cellular stress, has been identified as an Mst1-interacting protein, in a yeast two-hybrid screen of human adult prostate cDNA library with a dominant-negative Mst1 (K59R) as bait. The interaction of Mst1 with Hsp70 was confirmed by coimmunoprecipitation in both cotransfected HEK293 cells and prostate cancer cells. Hsp70 colocalized with Mst1 in the cytoplasm of LNCaP cells. The interaction sites with Mst1 consisted of NH(2)-terminal ATPase domain in Hsp70, whereas the inhibitory domain of Mst1 mediates the binding of Hsp70 in Mst1. Overexpression of Hsp70 mediates proteasomal degradation of Mst1 in a Hsp70 interacting protein (CHIP)-dependent manner. Furthermore, the proapoptotic effect of Mst1 was markedly inhibited by overexpression of Hsp70 or CHIP. Most strikingly, in response to the treatment of anticancer drug cisplatin, the induction of Hsp70 expression is higher in the androgen-independent DU145 cells compared with the androgen-dependent LNCaP cells. The higher levels of Hsp70 induction and subsequent Mst1 degradation mediate cisplatin resistance in prostate cancer DU145 cells. Moreover, overexpression of Mst1 sensitizes prostate cancer cells to cisplatin treatment. These findings implicate that Mst1, a downstream target of Hsp70, may be developed as a target for sensitizing hormone refractory prostate cancers to chemotherapy. PMID- 18381434 TI - A novel lung cancer signature mediates metastatic bone colonization by a dual mechanism. AB - Bone is a frequent target of lung cancer metastasis, which is associated with significant morbidity and a dismal prognosis. To identify and functionally characterize genes involved in the mechanisms of osseous metastasis, we developed a murine lung cancer model. Comparative transcriptomic analysis identified genes encoding signaling molecules (such as TCF4 and PRKD3) and cell anchorage-related proteins (MCAM and SUSD5), some of which were basally modulated by transforming growth factor-beta (TGF-beta) in tumor cells and in conditions mimicking tumor stromal interactions. Triple gene combinations induced not only high osteoclastogenic activity but also a marked enhancement of global metalloproteolytic activities in vitro. These effects were strongly associated with robust bone colonization in vivo, whereas this gene subset was ineffective in promoting local tumor growth and cell homing activity to bone. Interestingly, global inhibition of metalloproteolytic activities and simultaneous TGF-beta blockade in vivo led to increased survival and a remarkable attenuation of bone tumor burden and osteolytic metastasis. Thus, this metastatic gene signature mediates bone matrix degradation by a dual mechanism of induction of TGF-beta dependent osteoclastogenic bone resorption and enhancement of stroma-dependent metalloproteolytic activities. Our findings suggest the cooperative contribution of host-derived and cell autonomous effects directed by a small subset of genes in mediating aggressive osseous colonization. PMID- 18381435 TI - Detection of early prostate cancer using a hepsin-targeted imaging agent. AB - Early detection and diagnosis of prostate cancer is key to designing effective treatment strategies. Microarrays have resulted in the discovery of hepsin (HPN) as a biomarker for detection of prostate cancer. In this study, we explore the development of HPN imaging probes for detection of prostate cancer. We used phage display to isolate HPN binding peptides with 190 + 2.2 nmol/L affinity in monomeric form and high specificity. The identified peptides were able to detect human prostate cancer on tissue microarrays and in cell-based assays. HPN targeted imaging agents were synthesized by conjugating multiple peptides to fluorescent nanoparticles to further improve avidity through multivalency and to improve pharmacokinetics. When injected into mouse xenograft models, HPN-targeted nanoparticles bound specifically to HPN-expressing LNCaP xenografts compared with non-HPN-expressing PC3 xenografts. HPN imaging may provide a new method for detection of prostate cancer. PMID- 18381436 TI - TTI-237: a novel microtubule-active compound with in vivo antitumor activity. AB - 5-Chloro-6-[2,6-difluoro-4-[3-(methylamino)propoxy]phenyl]-N-[(1S)-2,2,2 trifluoro-1-methylethyl]-[1,2,4]triazolo[1,5-a]pyrimidin-7-amine butanedioate (TTI-237) is a microtubule-active compound of novel structure and function. Structurally, it is one of a class of compounds, triazolo[1,5a]pyrimidines, previously not known to bind to tubulin. Functionally, TTI-237 inhibited the binding of [(3)H]vinblastine to tubulin, but it caused a marked increase in turbidity development that more closely resembled the effect observed with docetaxel than that observed with vincristine. The morphologic character of the presumptive polymer is unknown at present. When applied to cultured human tumor cells at concentrations near its IC(50) value for cytotoxicity (34 nmol/L), TTI 237 induced multiple spindle poles and multinuclear cells, as did paclitaxel, but not vincristine or colchicine. Flow cytometry experiments revealed that, at low concentrations (20-40 nmol/L), TTI-237 produced sub-G(1) nuclei and, at concentrations above 50 nmol/L, it caused a strong G(2)-M block. The compound was a weak substrate of multidrug resistance 1 (multidrug resistance transporter or P glycoprotein). In a cell line expressing a high level of P-glycoprotein, the IC(50) of TTI-237 increased 25-fold whereas those of paclitaxel and vincristine increased 806-fold and 925-fold, respectively. TTI-237 was not recognized by the MRP or MXR transporters. TTI-237 was active in vivo in several nude mouse xenograft models of human cancer, including LoVo human colon carcinoma and U87-MG human glioblastoma, when dosed i.v. or p.o. Thus, TTI-237 has a set of properties that distinguish it from other classes of microtubule-active compounds. PMID- 18381437 TI - Blood vessel maturation and response to vascular-disrupting therapy in single vascular endothelial growth factor-A isoform-producing tumors. AB - Tubulin-binding vascular-disrupting agents (VDA) are currently in clinical trials for cancer therapy but the factors that influence tumor susceptibility to these agents are poorly understood. We evaluated the consequences of modifying tumor vascular morphology and function on vascular and therapeutic response to combretastatin-A4 3-O-phosphate (CA-4-P), which was chosen as a model VDA. Mouse fibrosarcoma cell lines that are capable of expressing all vascular endothelial growth factor (VEGF) isoforms (control) or only single isoforms of VEGF (VEGF120, VEGF164, or VEGF188) were developed under endogenous VEGF promoter control. Once tumors were established, VEGF isoform expression did not affect growth or blood flow rate. However, VEGF188 was uniquely associated with tumor vascular maturity, resistance to hemorrhage, and resistance to CA-4-P. Pericyte staining was much greater in VEGF188 and control tumors than in VEGF120 and VEGF164 tumors. Vascular volume was highest in VEGF120 and control tumors (CD31 staining) but total vascular length was highest in VEGF188 tumors, reflecting very narrow vessels forming complex vascular networks. I.v. administered 40 kDa FITC-dextran leaked slowly from the vasculature of VEGF188 tumors compared with VEGF120 tumors. Intravital microscopy measurements of vascular length and RBC velocity showed that CA-4-P produced significantly more vascular damage in VEGF120 and VEGF164 tumors than in VEGF188 and control tumors. Importantly, this translated into a similar differential in therapeutic response, as determined by tumor growth delay. Results imply differences in signaling pathways between VEGF isoforms and suggest that VEGF isoforms might be useful in vascular-disrupting cancer therapy to predict tumor susceptibility to VDAs. PMID- 18381438 TI - The cyclin-dependent kinase inhibitor flavopiridol potentiates the effects of topoisomerase I poisons by suppressing Rad51 expression in a p53-dependent manner. AB - The results of a phase I clinical trial of the topoisomerase I (Topo I) poison CPT-11 followed by the cyclin-dependent kinase inhibitor flavopiridol in patients with advanced solid tumors indicate that patients whose tumors were wild-type, but not mutant, for p53 obtained the most clinical benefit from this combination therapy. We elected to elucidate the mechanistic basis for this effect in isogenic-paired HCT116 colon cancer cells that were either wild-type (+/+) or null (-/-) for p53. With the combination therapy of SN-38 (the active metabolite of CPT-11) followed by flavopiridol, the induction of apoptosis was 5-fold greater in the p53+/+ cells compared with the p53-/- cells. This sequential treatment induced phosphorylation of p53 at Ser(15), which interacted with Rad51, a DNA repair protein involved in homologous recombination. Rad51 bound to p53 Ser(15) within the first 5 hours of combination therapy, and then was transcriptionally suppressed at 24 hours by flavopiridol only in p53+/+ cells. Microarray analysis also revealed suppression of Rad51 in a p53-dependent manner. Depletion of Rad51 by small interfering RNA (siRNA) sensitized both p53+/+ and p53-/- cells to SN-38-induced apoptosis with increase of gamma H2AX, a marker of DNA damage. Conversely, overexpression of Rad51 rescued p53+/+ cells from SN-->F induced apoptosis. Because flavopiridol inhibits Cdk9, we found that inhibition of Cdk9 by DRB or by siRNA could recapitulate the flavopiridol effects, with suppression of Rad51 and induction of apoptosis only in p53+/+ cells. In conclusion, after DNA damage by Topo I poisons, flavopiridol targets homologous recombination through a p53-dependent down-regulation of Rad51, resulting in enhancement of apoptosis. PMID- 18381439 TI - Therapeutic efficacy of ABT-737, a selective inhibitor of BCL-2, in small cell lung cancer. AB - Bcl-2 is a central regulator of cell survival that is overexpressed in the majority of small cell lung cancers (SCLC) and contributes to both malignant transformation and therapeutic resistance. We compared primary SCLC xenografts prepared from de novo human tumors with standard cell line-based xenografts in the evaluation of a novel and highly potent small molecule inhibitor of Bcl-2, ABT-737. ABT-737 induced dramatic regressions in tumors derived from some SCLC cell lines. In contrast, only one of three primary xenograft SCLC tumors showed significant growth inhibition with ABT-737. Explanations for this apparent dichotomy may include relatively low expression of Bcl-2 in the primary xenografts or inherent differences in the model systems. The addition of etoposide to ABT-737 in the primary xenografts resulted in significant decreases in tumor growth, underscoring the clinical potential of ABT-737 in combination therapy. To identify factors that may contribute to resistance to ABT-737 and related inhibitors, we isolated resistant derivatives of an initially sensitive cell line-based xenograft. Acquired resistance in this model was associated with decreases in the expression of the primary target Bcl-2, of proapoptotic partners of Bcl-2 (Bax and Bim), and of Bcl-2:Bim heterodimers. Expression profiling reveals 85 candidate genes demonstrating consistent changes in gene expression with acquired resistance. Taken together, these data have specific implications for the clinical development of Bcl-2 inhibitors for SCLC and broader implications for the testing of novel anticancer strategies in relevant preclinical models. PMID- 18381441 TI - FGFR2-amplified gastric cancer cell lines require FGFR2 and Erbb3 signaling for growth and survival. AB - We have identified a critical role for amplified FGFR2 in gastric cancer cell proliferation and survival. In a panel of gastric cancer cell lines, fibroblast growth factor receptor 2 (FGFR2) was overexpressed and tyrosine phosphorylated selectively in FGFR2-amplified cell lines KatoIII, Snu16, and OCUM-2M. FGFR2 kinase inhibition by a specific small-molecule inhibitor resulted in selective and potent growth inhibition in FGFR2-amplified cell lines, resulting in growth arrest in KatoIII cells and prominent induction of apoptosis in both Snu16 and OCUM-2M cells. FGFR2-amplified cell lines also contained elevated phosphotyrosine in EGFR, Her2, and Erbb3, but the elevated phosphorylation in EGFR could not be inhibited by gefitinib or erlotinib. We show that the elevated EGFR, Her2, and Erbb3 phosphotyrosine is dependent on FGFR2, revealing EGFR family kinases to be downstream targets of amplified FGFR2. Moreover, shRNA to Erbb3 resulted in a loss of proliferation, confirming a functional role for the activated EGFR signaling pathway. These results reveal that both the FGFR2 and EGFR family signaling pathways are activated in FGFR2-amplified gastric cancer cell lines to drive cell proliferation and survival. Inhibitors of FGFR2 or Erbb3 signaling may have therapeutic efficacy in the subset of gastric cancers containing FGFR2 amplification. PMID- 18381440 TI - Loss of E-cadherin promotes ovarian cancer metastasis via alpha 5-integrin, which is a therapeutic target. AB - E-cadherin loss is frequently associated with ovarian cancer metastasis. Given that adhesion to the abdominal peritoneum is the first step in ovarian cancer dissemination, we reasoned that down-regulation of E-cadherin would affect expression of cell matrix adhesion receptors. We show here that inhibition of E cadherin in ovarian cancer cells causes up-regulation of alpha(5)-integrin protein expression and transcription. When E-cadherin was blocked, RMUG-S ovarian cancer cells were able to attach and invade more efficiently. This greater efficiency could, in turn, be inhibited both in vitro and in vivo with an alpha(5)beta(1)-integrin-blocking antibody. When E-cadherin is silenced, alpha(5) integrin is up-regulated through activation of an epidermal growth factor receptor/FAK/Erk1-mitogen-activated protein kinase-dependent signaling pathway and not through the canonical E-cadherin/beta-catenin signaling pathway. In SKOV 3ip1 ovarian cancer xenografts, which express high levels of alpha(5)-integrin, i.p. treatment with an alpha(5)beta(1)-integrin antibody significantly reduced tumor burden, ascites, and number of metastasis and increased survival by an average of 12 days when compared with IgG treatment (P < 0.0005). alpha(5) Integrin expression was detected by immunohistochemistry in 107 advanced stage ovarian cancers using a tissue microarray annotated with disease-specific patient follow-up. Ten of 107 tissues (9%) had alpha(5)-integrin overexpression, and 39% had some level of alpha(5)-integrin expression. The median survival for patients with high alpha(5)-integrin levels was 26 months versus 35 months for those with low integrin expression (P < 0.05). Taken together, we have identified alpha(5) integrin up-regulation as a molecular mechanism by which E-cadherin loss promotes tumor progression, providing an explanation for how E-cadherin loss increases metastasis. Targeting this integrin could be a promising therapy for a subset of ovarian cancer patients. PMID- 18381442 TI - Two distinct molecular mechanisms underlying cytarabine resistance in human leukemic cells. AB - To understand the mechanism of cellular resistance to the nucleoside analogue cytarabine (1-beta-D-arabinofuranosylcytosine, AraC), two resistant derivatives of the human leukemic line CCRF-CEM were obtained by stepwise selection in different concentrations of AraC. CEM/4xAraC cells showed low AraC resistance, whereas CEM/20xAraC cells showed high resistance. Both cell lines showed similar patterns of cross-resistance to multiple cytotoxic nucleoside analogues, with the exception that CEM/20xAraC cells remained sensitive to 5-fluorouridine and 2 deoxy-5-fluorouridine. Both cell lines were sensitive to 5-fluorouracil and to a variety of natural product drugs. Although both CEM/4xAraC and CEM/20xAraC cells displayed reduced intracellular accumulation of [(3)H]AraC, only CEM/4xAraC cells showed reduced uptake of [(3)H]uridine, which was used to assess nucleoside transport activities. Genes encoding proteins known to be involved in nucleoside transport, efflux, and metabolism were analyzed for the presence of mutations in the two cell lines. In CEM/4xAraC cells, independent mutations were identified at each allele of human equilibrative nucleoside transporter 1 (hENT1; SLC29A1), one corresponding to a single-nucleotide change in exon 4, the other being a complex intronic mutation disrupting splicing of exon 13. In contrast to CEM/20xAraC cells, CEM/4xAraC cells did not bind the hENT1/SLC29A1 ligand nitrobenzylmercaptopurine ribonucleoside and lacked detectable hENT1/SLC29A1 protein. In CEM/20xAraC cells, independent intronic mutations impairing splicing of exons 2 and 3 were found at each allele of the deoxycytidine kinase gene. These studies point to at least two distinct mechanisms of AraC resistance in leukemic cells. PMID- 18381443 TI - The nucleolin targeting aptamer AS1411 destabilizes Bcl-2 messenger RNA in human breast cancer cells. AB - We sought to determine whether nucleolin, a bcl-2 mRNA-binding protein, has a role in the regulation of bcl-2 mRNA stability in MCF-7 and MDA-MB-231 breast cancer cells. Furthermore, we examined the efficacy of the aptamer AS1411 in targeting nucleolin and inducing bcl-2 mRNA instability and cytotoxicity in these cells. AS1411 at 5 micromol/L inhibited the growth of MCF-7 and MDA-MB-231 cells, whereas 20 micromol/L AS1411 had no effect on the growth rate or viability of normal MCF-10A mammary epithelial cells. This selectivity of AS1411 was related to a greater uptake of AS1411 into the cytoplasm of MCF-7 cells compared with MCF 10A cells and to a 4-fold higher level of cytoplasmic nucleolin in MCF-7 cells. Stable siRNA knockdown of nucleolin in MCF-7 cells reduced nucleolin and bcl-2 protein levels and decreased the half-life of bcl-2 mRNA from 11 to 5 hours. Similarly, AS1411 (10 micromol/L) decreased the half-life of bcl-2 mRNA in MCF-7 and MDA-MB-231 cells to 1.0 and 1.2 hours, respectively. In contrast, AS1411 had no effect on the stability of bcl-2 mRNA in normal MCF-10A cells. AS1411 also inhibited the binding of nucleolin to the instability element AU-rich element 1 of bcl-2 mRNA in a cell-free system and in MCF-7 cells. Together, the results suggest that AS1411 acts as a molecular decoy by competing with bcl-2 mRNA for binding to cytoplasmic nucleolin in these breast cancer cell lines. This interferes with the stabilization of bcl-2 mRNA by nucleolin and may be one mechanism by which AS1411 induces tumor cell death. PMID- 18381444 TI - Characterization of an Akt kinase inhibitor with potent pharmacodynamic and antitumor activity. AB - Akt kinases 1, 2, and 3 are important regulators of cell survival and have been shown to be constitutively active in a variety of human tumors. GSK690693 is a novel ATP-competitive, low-nanomolar pan-Akt kinase inhibitor. It is selective for the Akt isoforms versus the majority of kinases in other families; however, it does inhibit additional members of the AGC kinase family. It causes dose dependent reductions in the phosphorylation state of multiple proteins downstream of Akt, including GSK3 beta, PRAS40, and Forkhead. GSK690693 inhibited proliferation and induced apoptosis in a subset of tumor cells with potency consistent with intracellular inhibition of Akt kinase activity. In immune compromised mice implanted with human BT474 breast carcinoma xenografts, a single i.p. administration of GSK690693 inhibited GSK3 beta phosphorylation in a dose- and time-dependent manner. After a single dose of GSK690693, >3 micromol/L drug concentration in BT474 tumor xenografts correlated with a sustained decrease in GSK3 beta phosphorylation. Consistent with the role of Akt in insulin signaling, treatment with GSK690693 resulted in acute and transient increases in blood glucose level. Daily administration of GSK690693 produced significant antitumor activity in mice bearing established human SKOV-3 ovarian, LNCaP prostate, and BT474 and HCC-1954 breast carcinoma xenografts. Immunohistochemical analysis of tumor xenografts after repeat dosing with GSK690693 showed reductions in phosphorylated Akt substrates in vivo. These results support further evaluation of GSK690693 as an anticancer agent. PMID- 18381445 TI - Statins increase p21 through inhibition of histone deacetylase activity and release of promoter-associated HDAC1/2. AB - Statins are 3-hydroxy-3-methylglutaryl-CoA reductase inhibitors broadly used for the control of hypercholesterolemia. Recently, they are reported to have beneficial effects on certain cancers. In this study, we show that statins inhibited the histone deacetylase (HDAC) activity and increased the accumulation of acetylated histone-H3 and the expression of p21(WAF/CIP) in human cancer cells. Computational modeling showed the direct interaction of the carboxylic acid moiety of statins with the catalytic site of HDAC2. In the subsequent enzymatic assay, it was shown that lovastatin inhibited HDAC2 activity competitively with a K(i) value of 31.6 micromol/L. Sp1 but not p53 sites were found to be the statins-responsive element shown by p21 luciferase-promoter assays. DNA affinity protein binding assay and chromatin immunoprecipitation assay showed the dissociation of HDAC1/2 and association of CBP, leading to the histone-H3 acetylation on the Sp1 sites of p21 promoter. In vitro cell proliferation and in vivo tumor growth were both inhibited by statins. These results suggest a novel mechanism for statins through abrogation of the HDAC activity and promoter histone-H3 acetylation to regulate p21 expression. Therefore, statins might serve as novel HDAC inhibitors for cancer therapy and chemoprevention. PMID- 18381446 TI - Mammalian target of rapamycin and S6 kinase 1 positively regulate 6-thioguanine induced autophagy. AB - DNA mismatch repair (MMR) ensures the fidelity of DNA replication and is required for activation of cell cycle arrest and apoptosis in response to certain classes of DNA damage. We recently reported that MMR is also implicated in initiation of an autophagic response after MMR processing of 6-thioguanine (6-TG). It is now generally believed that autophagy is negatively controlled by mammalian target of rapamycin (mTOR) activity. To determine whether mTOR is involved in 6-TG-induced autophagy, we used rapamycin, a potential anticancer agent, to inhibit mTOR activity. Surprisingly, we find that rapamycin cotreatment inhibits 6-TG-induced autophagy in MMR-proficient human colorectal cancer HCT116 (MLH1(+)) and HT29 cells as measured by LC3 immunoblotting, GFP-LC3 relocalization, and acridine orange staining. Consistently, short interfering RNA silencing of the 70-kDa ribosomal S6 kinase 1 (S6K1), the downstream effector of mTOR, markedly reduces 6 TG-induced autophagy. Furthermore, we show that inhibition of mTOR by rapamycin induces the activation of Akt as shown by increased Akt phosphorylation at Ser(473) and the inhibition of 6-TG-induced apoptosis and cell death. Activated Akt is a well-known inhibitor of autophagy. In conclusion, our data indicate that mTOR-S6K1 positively regulates autophagy after MMR processing of 6-TG probably through its negative feedback inhibition of Akt. PMID- 18381447 TI - Epithelial-to-mesenchymal transition and integrin-linked kinase mediate sensitivity to epidermal growth factor receptor inhibition in human hepatoma cells. AB - Hepatocellular carcinoma (HCC) is associated with a poor prognosis due to late diagnoses and a lack of effective treatment options. Epidermal growth factor receptor (EGFR)-targeted therapies have been effective in other cancers. However, erlotinib and cetuximab have shown only modest efficacy in clinical trials of HCC. We examined epithelial-to-mesenchymal transition (EMT) as a determinant of sensitivity of HCC to EGFR inhibitors. A panel of 12 human hepatoma cell lines were classified as epithelial or mesenchymal based on their expression of E cadherin and vimentin. The resulting classification correlated with a previous microarray analysis of human hepatoma cell lines whereby the mesenchymal cell lines were shown to have increased expression of genes involved in metastasis and invasion. Sensitivity to erlotinib, gefitinib, and cetuximab was assessed and the epithelial cell lines were found to be significantly more susceptible to all three agents. Analysis of the EGFR pathway showed that EMT status was independent of EGFR expression or downstream extracellular signal-regulated kinase activation and only the epithelial cell lines expressed ErbB3. Interestingly, mesenchymal cells resistant to EGFR inhibitors had increased AKT and signal transducer and activator of transcription-3 activation through elevated expression of integrin linked kinase (ILK). Mesenchymal cell lines were therefore experimentally transformed with kinase-inactive ILK (KI-ILK) with a resulting decrease in ILK activity and activation of AKT. KI-ILK transformants showed increased sensitivity to EGFR inhibitors both in vitro and in an in vivo xenograft model. These data suggest that EMT predicts HCC sensitivity to EGFR-targeted therapies and that ILK is a novel target to overcome HCC resistance to EGFR inhibition. PMID- 18381448 TI - Development of novel tetravalent anti-CD20 antibodies with potent antitumor activity. AB - Despite the effectiveness of the anti-CD20 monoclonal antibody (mAb) Rituximab (C2B8) in the treatment of B-cell lymphoma, its efficacy remains variable and often modest. It seems likely that a combination of multiple mechanisms, such as complement-dependent cytotoxicity (CDC) and apoptotic signaling, underlies the therapeutic success of anti-CD20 mAbs. Unfortunately, all the current anti-CD20 mAbs effective in CDC are relatively inactive in signaling cell death and vice versa. In this study, we developed two genetically engineered tetravalent antibodies (TetraMcAb) respectively derived from the anti-CD20 mAbs C2B8 and 2F2. TetraMcAbs, with a molecular mass only 25 kDa higher than native divalent antibodies (DiMcAb), were shown not only to be as effective in mediating CDC and antibody-dependent cellular cytotoxicity against B-lymphoma cells as DiMcAbs but also to have antiproliferative and apoptosis-inducing activity markedly superior to that of DiMcAbs. Interestingly, whereas 2F2 and C2B8 were equally effective in inducing cell growth arrest and apoptosis, the functions of their tetravalent versions, 2F2(ScFvHL)(4)-Fc and C2B8(ScFvHL)(4)-Fc, were significantly different. 2F2(ScFvHL)(4)-Fc exhibited exceptionally more potent antiproliferative and apoptosis-inducing activity than that of C2B8(ScFvHL)(4)-Fc. Immunotherapeutic studies further showed that 2F2(ScFvHL)(4)-Fc was far more effective in prolonging the survival of severe combined immunodeficient mice bearing systemic Daudi or Raji tumors than C2B8, 2F2, and C2B8(ScFvHL)(4)-Fc, suggesting that it might be a promising therapeutic agent for B-cell lymphoma. PMID- 18381450 TI - MCF7 side population cells with characteristics of cancer stem/progenitor cells express the tumor antigen MUC1. AB - Chemotherapy, radiation, and growth inhibitory drugs preferentially eliminate actively growing cancer cells. Cancer recurrence is currently thought to be due to nondividing cancer stem/progenitor cells that are resistant to these therapies. Different therapeutic approaches need to be considered for the elimination of the cancer stem cell population. Immunotherapy is one such approach. In addition to specificity and lack of toxicity, immunotherapy targets cancer cells irrespective of their state of proliferation, as long as they express particular tumor antigens. For that reason, it is important to examine if the tumor antigens that are currently being tested as immunotherapeutic agents are also present on cancer stem cells. This study aimed to determine if one well known tumor antigen, MUC1, which is being tested as an immunotherapy target on tumor cells, is also expressed on the quiescent cancer stem/progenitor cells. We used the so-called side population (SP) cells found in the MCF7 breast cancer cell line, which we first confirmed by cell surface markers and gene profiling to be highly enriched in cells that fulfill specific functional, phenotypic, and molecular criteria for being tumor stem/progenitor cells. We show that these cells express MUC1 and give rise to MUC1(+) tumors in vivo, which maintain the MUC1(+) SP population. MUC1 on SP cells is hypoglycosylated and heavily sialylated; the characteristics of the tumor-specific form were expressed on mature cancer cells and recognized by tumor-specific T cells and antibodies. This suggests that stem/progenitor cells, like mature tumor cells, would be targets of MUC1-directed immunotherapy. PMID- 18381449 TI - Conjugation of human topoisomerase 2 alpha with small ubiquitin-like modifiers 2/3 in response to topoisomerase inhibitors: cell cycle stage and chromosome domain specificity. AB - Type 2 topoisomerases, in particular the alpha isoform in human cells, play a key role in cohesion and sister chromatid separation during mitosis. These enzymes are thus vital for cycling cells and are obvious targets in cancer chemotherapy. Evidence obtained in yeast and Xenopus model systems indicates that conjugation of topoisomerase 2 with small ubiquitin-like modifier (SUMO) proteins is required for its mitotic functions. Here, we provide biochemical and cytologic evidence that topoisomerase 2 alpha is conjugated to SUMO-2/3 during interphase and mitosis in response to topoisomerase 2 inhibitors and "poisons" (ICRF-187, etoposide, doxorubicin) that stabilize catalytic intermediates (cleavage complexes, closed clamp forms) of the enzyme onto target DNA. During mitosis, SUMO-2/3-modified forms of topoisomerase 2 alpha localize to centromeres and chromosome cores/axes. However, centromeres are unresponsive to inhibitors during interphase. Furthermore, formation of topoisomerase 2 alpha-SUMO-2/3 conjugates within mitotic chromosomes strongly correlates with incomplete chromatid decatenation and decreases progressively as cells approach the metaphase-anaphase transition. We also found that the PIASy protein, an E3 ligase for SUMO proteins, colocalizes with SUMO-2/3 at the mitotic chromosomal cores/axes and is necessary for both formation of SUMO-2/3 conjugates and proper chromatid segregation. We suggest that the efficacy of topoisomerase inhibitors to arrest cells traversing mitosis may relate to their targeting of topoisomerase 2 alpha-SUMO-2/3 conjugates that concentrate at mitotic chromosome axes and are directly involved in chromatid arm separation. PMID- 18381451 TI - Antitumor effects of Mucin 1/sec involves the modulation of urokinase-type plasminogen activator and signal transducer and activator of transcription 1 expression in tumor cells. AB - Expression of the transmembrane isoform of Mucin 1 (MUC1/TM) in an aggressive murine mammary tumor line, DA-3, does not alter tumor development and metastasis, leading to death of the host. However, tumor cells expressing a secreted isoform of MUC1 (MUC1/sec) fail to develop tumors in immunocompetent mice. The rejection of MUC1/sec-expressing tumor cells is immunologically mediated, as, initially, innate cells and, ultimately, T cells are required. After gene array analysis, and confirmation at the protein level, it was discovered that MUC1/sec-expressing tumor cells (DA-3/sec) have a significant reduction in expression of urokinase type plasminogen activator (uPA) relative to the parental tumor line and tumor cells expressing MUC1/TM. The serine protease uPA has been found to be involved in growth-promoting signaling, angiogenesis, and induction of matrix remodeling leading to metastasis. Although the tumor-promoting Stat3 transcription factor was unaltered in these tumor cells, the tumor-suppressive and IFN-responsive signal transducer and activator of transcription 1 (Stat1) is dramatically up regulated in DA-3/sec cells. In addition, treatment of various murine and human cell lines with conditioned medium containing MUC1/sec results in up-regulation of Stat1. DA-3/sec tumor cells are also sensitized to the antiproliferative effects of IFN-gamma. Furthermore, transfection of the Stat1 gene into DA-3 tumor cells leads to a down-regulation of uPA and delays tumor progression. Thus, Stat1 up-regulation in DA-3/sec cells seems to play a significant role in the mechanism(s) by which rejection of tumor cells expressing MUC1/sec may be occurring. PMID- 18381453 TI - A cryptic vascular endothelial growth factor T-cell epitope: identification and characterization by mass spectrometry and T-cell assays. AB - Vascular endothelial growth factor (VEGF) is involved in various physiologic processes, such as angiogenesis or wound healing, but is also crucial in pathologic events, such as tumor growth. Thus, clinical anti-VEGF treatments have been developed that could already show beneficial effects for cancer patients. In this article, we describe the first VEGF-derived CD8(+) T-cell epitope. The natural HLA ligand SRFGGAVVR was identified by differential mass spectrometry in two primary renal cell carcinomas (RCC) and was significantly overpresented on both tumor tissues. SRFGGAVVR is derived from a cryptic translated region of VEGF presumably by initiation of translation at the nonclassic start codon CUG(499). SRFGGAVVR-specific T cells were generated in vitro using peptide-loaded dendritic cells or artificial antigen-presenting cells. SRFGGAVVR-specific CD8(+) T cells, identified by HLA tetramer analysis after in vitro stimulation, were fully functional T effector cells, which were able to secrete IFN-gamma on stimulation and killed tumor cells in vitro. Additionally, we have quantitatively analyzed VEGF mRNA and protein levels in RCC tumor and normal tissue samples by gene chip analysis, quantitative reverse transcription-PCR, in situ hybridization, and bead based immunoassay. In the future, T cells directed against VEGF as a tumor associated antigen may represent a possible way of combining peptide-based anti VEGF immunotherapy with already existent anti-VEGF cancer therapies. PMID- 18381452 TI - Signatures associated with rejection or recurrence in HER-2/neu-positive mammary tumors. AB - We have previously shown T-cell-mediated rejection of the neu-overexpressing mammary carcinoma cells (MMC) in wild-type FVB mice. However, following rejection of primary tumors, a fraction of animals experienced a recurrence of a neu antigen-negative variant (ANV) of MMC (tumor evasion model) after a long latency period. In the present study, we determined that T cells derived from wild-type FVB mice can specifically recognize MMC by secreting IFN-gamma and can induce apoptosis of MMC in vitro. Neu transgenic (FVBN202) mice develop spontaneous tumors and cannot reject it (tumor tolerance model). To dissect the mechanisms associated with rejection or tolerance of MMC tumors, we compared transcriptional patterns within the tumor microenvironment of MMC undergoing rejection with those that resisted it either because of tumor evasion/antigen loss recurrence (ANV tumors) or because of intrinsic tolerance mechanisms displayed by the transgenic mice. Gene profiling confirmed that immune rejection is primarily mediated through activation of IFN-stimulated genes and T-cell effector mechanisms. The tumor evasion model showed combined activation of Th1 and Th2 with a deviation toward Th2 and humoral immune responses that failed to achieve rejection likely because of lack of target antigen. Interestingly, the tumor tolerance model instead displayed immune suppression pathways through activation of regulatory mechanisms that included in particular the overexpression of interleukin-10 (IL 10), IL-10 receptor, and suppressor of cytokine signaling (SOCS)-1 and SOCS-3. These data provide a road map for the identification of novel biomarkers of immune responsiveness in clinical trials. PMID- 18381454 TI - Skewing the T-cell repertoire by combined DNA vaccination, host conditioning, and adoptive transfer. AB - Approaches for T-cell-based immunotherapy that have shown substantial effects in clinical trials are generally based on the adoptive transfer of high numbers of antigen-specific cells, and the success of these approaches is thought to rely on the high magnitude of the tumor-specific T-cell responses that are induced. In this study, we aimed to develop strategies that also yield a T-cell repertoire that is highly skewed toward tumor recognition but do not rely on ex vivo generation of tumor-specific T cells. To this end, the tumor-specific T-cell repertoire was first expanded by DNA vaccination and then infused into irradiated recipients. Subsequent vaccination of the recipient mice with the same antigen resulted in peak CD8(+) T-cell responses of approximately 50%. These high T-cell responses required the presence of antigen-experienced tumor-specific T cells within the graft because only mice that received cells of previously vaccinated donor mice developed effective responses. Tumor-bearing mice treated with this combined therapy showed a significant delay in tumor outgrowth, compared with mice treated by irradiation or vaccination alone. Furthermore, this antitumor effect was accompanied by an increased accumulation of activated and antigen specific T cells within the tumor. In summary, the combination of DNA vaccination with host conditioning and adoptive transfer generates a marked, but transient, skewing of the T-cell repertoire toward tumor recognition. This strategy does not require ex vivo expansion of cells to generate effective antitumor immunity and may therefore easily be translated to clinical application. PMID- 18381455 TI - Immunity to growth factor receptor-bound protein 10, a signal transduction molecule, inhibits the growth of breast cancer in mice. AB - This study describes the application of a unique strategy to identify breast cancer antigens [tumor-associated antigen (TAA)]. In a mouse model, the strategy led to the identification of growth factor receptor-bound protein 10 (Grb10) as a newly identified TAA. Grb10 is a signal transduction molecule associated with multiple transmembrane tyrosine kinase receptors. It was discovered by comparing microarrays of cellular breast cancer vaccines highly enriched for cells that induced breast cancer immunity in tumor-bearing mice with nonenriched vaccines. The vaccines were prepared by transferring a cDNA expression library derived from SB5b cells, a breast cancer cell line C3H/He origin (H-2(k)), into LM mouse fibroblasts (H-2(k)). As the transferred cDNA integrates spontaneously into the genome of the recipient cells, replicates as the cells divide, and is expressed, the vaccine could be prepared from microgram amounts of tumor tissue. Relatively few cells in the transduced cell population, however, incorporated cDNA fragments that included genes specifying TAA. (The vast majority specified normal cellular constituents.) A unique strategy was used, therefore, to enrich the vaccine for immunotherapeutic cells. Twenty genes were overrepresented in the enriched vaccines. One, the gene for Grb10, was approximately 100-fold overrepresented. To determine if Grb10 in the enriched vaccine was partly responsible for its therapeutic benefits, the gene was transferred into the fibroblast cell line, which was then used as a vaccine. Mice with established breast cancer treated solely by immunization with the modified fibroblasts developed robust immunity to the breast cancer cells, which, in some instances, was sufficient to result in tumor rejection. PMID- 18381456 TI - An effective vaccine strategy protective against antigenically distinct tumor variants. AB - Antigenically distinct tumor variants can emerge in response to selective pressures inherent to host-tumor interactions. The development of successful immunotherapeutic strategies can be limited by these disparate antigenic profiles. Using the immunomodulator B7-DC XAb to activate cytolytic T cells specific for tumor-associated antigens, we found that the specificity of immune responses elicited by live tumors are distinct from the specificity of the responses elicited by soluble proteins derived from the same tumors. Remarkably, whereas the induced antitumor immunity generated against live variants of the B16 melanoma and EL4 thymic lymphoma tumors were highly specific for the original tumor variant used in the challenge, immunity generated using soluble proteins derived from tumor lysates was broadly reactive, recognizing the challenge tumor, as well as antigenically distinct variants. The antigens detected using live tumor and tumor lysate vaccines could be distinguished biochemically, demonstrating that they are structurally distinct. We show that vaccines using antigens present in tumor cell lysates induce protective immunity with strong memory against distantly related tumor variants. The existence of a class of antigens shared among tumor variants provides an attractive target for vaccine development. PMID- 18381457 TI - Insulin-like growth factor-I-dependent up-regulation of ZEB1 drives epithelial-to mesenchymal transition in human prostate cancer cells. AB - The epithelial-to-mesenchymal transition (EMT) is crucial for the migration and invasion of many epithelial tumors, including prostate cancer. Although it is known that ZEB1 overexpression promotes EMT primarily through down-regulation of E-cadherin in a variety of cancers, the soluble ligands responsible for the activation of ZEB1 have yet to be identified. In the present study, we investigated the role of insulin-like growth factor-I (IGF-I) in the regulation of ZEB1 during EMT associated with prostate tumor cell migration. We found that ZEB1 is expressed in highly aggressive prostate cancer cells and that its expression correlates directly with Gleason grade in human prostate tumors (P < 0.001). IGF-I up-regulates ZEB1 expression in prostate cancer cells exhibiting an epithelial phenotype. In prostate cancer cells displaying a mesenchymal phenotype, ZEB1 inhibition reverses the suppression of E-cadherin protein and down-regulates the expression of the mesenchymal markers N-cadherin and fibronectin. Furthermore, ZEB1 blockade decreases migratory and invasive potential in ARCaP(M) compared with the control. These results identify ZEB1 as a key transcriptional regulator of EMT in prostate cancer and suggest that the aberrant expression of ZEB1 in prostate cancer cells occurs in part in response to IGF-I stimulation. PMID- 18381458 TI - Identification of novel methylation markers in cervical cancer using restriction landmark genomic scanning. AB - Aberrant methylation of CpG islands in gene promoters often represents an early clonal event in carcinogenesis. Accordingly, defining methylation profiles may be useful for developing marker panels for early detection or predicting the risk of cancer precursors. To identify specific genes frequently methylated in cervical cancer, we conducted methylation profiling of 20 primary human cervical cancers using NotI-based restriction landmark genomic scanning (RLGS). Of 2,172 RLGS fragments analyzed (average, 1,753 CpG islands per patient), 186 RLGS fragments were lost in at least one tumor and 40 were lost in three or more. Methylation was identified in 19 (95%) of 20 tumor samples compared with normal DNA. Bisulfite sequencing was conducted to confirm RLGS results. Of the confirmed markers frequently methylated, we developed Methylight assays for two corresponding genes, nucleolar protein 4 (NOL4), and lipoma HMGIC fusion partner like protein 4 (LHFPL4), which were methylated in 85% and 55% of cancers, respectively. Using these assays, we further confirmed frequent CpG island methylation in the original cancers and in another independent series of 15 cervical cancers. We also showed methylation at a reduced frequency in a set of carefully reviewed cytology specimens demonstrating cells exfoliated from cancer precursor lesions. In summary, we identified, for the first time, NOL4 and LHFPL4 as novel methylation targets specific for cervical cancer. Inclusion of NOL4 and LHFPL4 in evaluating methylation panels for early detection, risk prediction, and etiologic research on cervical cancer is warranted. PMID- 18381459 TI - Genetic variation in the one-carbon transfer pathway and ovarian cancer risk. AB - Dysfunction in enzymes involved in one-carbon (1-C) metabolism can lead to increased chromosomal strand breaking and abnormal methylation patterns, which are both associated with cancer risk. Availability of 1-C units may modify risk. We investigated the association of single-nucleotide polymorphisms (SNP) in 21 genes in the 1-C transfer pathway among 829 Caucasian cases with primary epithelial ovarian cancer and 941 frequency-matched unaffected controls enrolled at Mayo Clinic (Rochester, MN) and Duke University (Durham, NC) and examined risk modification by multivitamin supplement use. Multivariable-adjusted SNP-specific logistic regression and haplotype analyses were done for 180 SNPs and false positive report probabilities (FPRP) were calculated. Each copy of the minor allele in SHMT1 intron 5 A>G (rs9909104) was associated with epithelial ovarian cancer [odds ratio (OR), 1.2; 95% confidence interval (95% CI), 1.0-1.4; P trend = 0.02; FPRP = 0.16] and a 5-SNP SHMT1 haplotype was associated with decreased risk (P = 0.01; FPRP = 0.09). Three SNPs in DNMT3A were associated with risk among multivitamin supplement users: 3' untranslated region (UTR) C>G (rs13420827: OR, 0.8; 95% CI, 0.6-1.0; P interaction = 0.006; FPRP = 0.54), intron 6 G>A (rs11887120: OR, 0.8; 95% CI, 0.7-1.0; P interaction = 0.007; FPRP = 0.57), and intron 22 A>T (rs11695471: OR, 1.2; 95% CI, 1.0-1.5; P interaction = 0.01; FPRP = 0.66). These data extend previous findings from other cancers of a role for SHMT1 in ovarian cancer, and provide evidence that SNPs in methylation and DNA synthesis reactions are associated with risk of ovarian cancer. Interventions with modifiable factors such as multivitamin intake may reduce risk. PMID- 18381460 TI - Aspirin, NSAID, and acetaminophen use and the risk of endometrial cancer. AB - To date, no prospective studies have explored the relationship between the use of aspirin, other nonsteroidal anti-inflammatory medications (NSAID), and acetaminophen and endometrial adenocarcinoma. Of the 82,971 women enrolled in a prospective cohort study, 747 developed medical record-confirmed invasive endometrial cancer over a 24-year period. Use of aspirin was ascertained from 1980 to 2004, and for other NSAIDs and acetaminophen, from 1990 to 2004. Cox regression models calculated multivariate relative risks (MV RR), controlling for body mass index (BMI), postmenopausal hormone (PMH) use, and other endometrial cancer risk factors. Currency, duration, and quantity of aspirin were not associated with endometrial cancer risk overall [current use: MV RR, 1.03; 95% confidence interval (CI) 0.83-1.27; >10 years of use: MV RR, 1.01; 95% CI, 0.78 1.30; and cumulative average >7 tablets per week: (MV RR, 1.10; 95% CI, 0.84 1.44)]. However, stratified analyses showed that a lower risk of endometrial cancer among obese (BMI, >or=30 kg/m(2)) women was seen with current aspirin use (MV RR, 0.66; 95% CI, 0.46-0.95). The greatest risk reduction for current aspirin users was seen in postmenopausal obese women who had never used PMH (MV RR, 0.43; 95% CI, 0.26-0.73). The use of other NSAIDs or acetaminophen was not associated with endometrial cancer. Our data suggest that use of aspirin or other NSAIDs does not play an important role in endometrial cancer risk overall. However, risk was significantly lower for current aspirin users who were obese or who were postmenopausal and had never used PMHs; these subgroup findings require further confirmation. PMID- 18381461 TI - The majority of viral-cellular fusion transcripts in cervical carcinomas cotranscribe cellular sequences of known or predicted genes. AB - Integration of human papillomavirus (HPV) DNA into the host genome is a frequent event in cervical carcinogenesis and is reported to occur at randomly selected chromosomal sites. However, as the databases are being up-dated continuously, the knowledge based on sequenced viral integration sites also expands. In this study, viral-cellular fusion transcripts of a preselected group of 74 cervical carcinoma or cervical intraepithelial neoplasia grade 3 (CIN3) biopsies harboring integrated HPV16, HPV18, HPV31, HPV33, or HPV45 DNA were amplified by 3'-rapid amplification of cDNA ends PCR and sequenced. Consistent with previous reports, integration sites were found to be distributed throughout the genome. However, 23% (17 of 74) of the integration sites were located within the cytogenetic bands 4q13.3, 8q24.21, 13q22.1, and 17q21, in clusters ranging from 86 to 900 kb. Of note is that clusters 8q24.21 and 13q22.1 are within 1.5 Mbp of an adjacent fragile site whereas clusters 4q13.3 and 17q21 are >15 Mbp distant to any known fragile sites. It is tempting to speculate that as yet unknown fragile sites may be identified on the basis of HPV integration hotspots. No correlation between HPV type and specific integration loci was found. Of 74 fusion transcripts, 28 contained cellular sequences, which were homologous to known genes, and 40 samples contained sequences of predicted genes. In 33 fusion transcripts, both viral and cellular sequences were in sense orientation, indicating that the gene itself or upstream sequences were affected by integration. These data suggest that the influence of HPV integration on host gene expression may not be a rare effect and should encourage more detailed analyses. PMID- 18381462 TI - Effect of caffeine on the ATR/Chk1 pathway in the epidermis of UVB-irradiated mice. AB - Administration of caffeine was shown in earlier studies to enhance UVB-induced apoptosis and inhibit UVB-induced carcinogenesis in hairless SKH-1 mice. Here, we describe a potential mechanism for these in vivo effects. A single irradiation of mouse skin with UVB activated the ataxia-telangiectasia mutated- and Rad3-related (ATR) pathway, causing a severalfold increase in keratinocytes with phospho-Chk1 (Ser(345)) and a marked decrease in mitotic keratinocytes with cyclin B1 compared with baseline. When given in the drinking water for 1 to 2 weeks before UVB, caffeine (0.4 mg/mL) markedly inhibited the UVB-induced phosphorylation of Chk1 on Ser(345) and caused premature expression of cyclin B1 in the epidermis. Normal keratinocytes had delayed mitotic entry for >10 h following UVB. Caffeine administration reduced this mitotic delay to only 4 h and caused markedly increased apoptosis by 6 to 10 h after UVB. p53 knockout mice were used to determine the role of p53 in these processes. Irradiation with UVB markedly decreased the number of mitotic keratinocytes with cyclin B1 in p53 knockout mice, and topical caffeine immediately after UVB abrogated this response and increased UVB-induced apoptosis severalfold. These effects of caffeine in knockout mice were substantially greater than in wild-type mice. The ability of caffeine to promote the deletion of p53(-/-) keratinocytes may be relevant to its inhibitory effect on UVB-induced skin cancer. Our studies indicate that administration of caffeine enhances the removal of DNA-damaged cells by inhibiting the ATR-mediated phosphorylation of Chk1 and prematurely increasing the number of cyclin B1-containing cells that undergo lethal mitosis. PMID- 18381463 TI - Evaluation of genetic variants in microRNA-related genes and risk of bladder cancer. AB - MicroRNAs (miRNA) are small noncoding RNA molecules involved in a diversity of cellular functions. Although it has been reported that global suppression of the miRNA biogenesis pathway leads to enhanced tumorigenesis, the effect of common genetic variants of miRNA-related genes on cancer predisposition is unclear. To better understand this effect, we genotyped 41 single-nucleotide polymorphisms (SNP) from 24 miRNA-related genes in a case-control study conducted in 746 Caucasian patients with bladder cancer and 746 matched controls. The homozygous variant genotype of a nonsynonymous SNP in the GEMIN3 gene (rs197414) was associated with a significantly increased bladder cancer risk [odds ratios (OR), 2.40; 95% confidence interval (95% CI), 1.04-5.56]. Several additional miRNA related SNPs were also identified that showed a borderline significant association with bladder cancer risk. Haplotype analysis indicated that a common haplotype of the GEMIN4 gene was associated with a significantly increased bladder cancer risk with an OR of 1.25 (95% CI, 1.01-1.54). To assess the aggregate effects of the promising SNPs, we performed a combined unfavorable genotype analysis that included all SNPs showing at least a borderline statistical significance. We found that, compared with the low-risk reference group with less than two unfavorable genotypes, the medium-risk group with two unfavorable genotypes exhibited a 1.29-fold (0.92-1.81) increased risk whereas the high-risk group with more than two unfavorable genotypes exhibited a 1.92 fold (1.36-2.71) increased risk (P(trend) < 0.0001). Overall, this is the first epidemiologic study showing that miRNA-related genetic variants may affect bladder cancer risk individually and jointly. PMID- 18381466 TI - Validation of colorectal cancer screening behaviors. PMID- 18381464 TI - Mitogen- and stress-activated kinase 1-mediated histone H3 phosphorylation is crucial for cell transformation. AB - Mitogen- and stress-activated kinase 1 (MSK1) belongs to a family of dual protein kinases that are activated by either extracellular signal-regulated kinase or p38 mitogen-activated protein kinases in response to stress or mitogenic extracellular stimuli. The physiologic role of MSK1 in malignant transformation and cancer development is not well understood. Here, we report that MSK1 is involved in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced or epidermal growth factor (EGF)-induced neoplastic transformation of JB6 Cl41 cells. H89, a potent inhibitor of MSK1, strongly suppressed TPA-induced or EGF-induced cell transformation. When cells overexpressing wild-type MSK1 were treated with TPA or EGF, colony formation increased substantially compared with untreated cells or cells that did not overexpress MSK1. In contrast, MSK1 COOH terminal or NH(2) terminal dead dominant negative mutants dramatically suppressed cell transformation. Introduction of small interfering RNA-MSK1 into JB6 Cl41 cells resulted in suppressed TPA-induced or EGF-induced cell transformation. In addition, cell proliferation was inhibited in MSK1 knockdown cells compared with MSK1 wild-type cells. In wild-type MSK1-overexpressing cells, activator protein (AP-1) activation increased after TPA or EGF stimulation, whereas AP-1 activation decreased in both MSK1 dominant-negative mutants and in MSK1 knockdown cells. Moreover, TPA-induced or EGF-induced phosphorylation of histone H3 at Ser(10) was increased in wild-type cells but the induced phosphorylation was abolished in MSK1 dominant-negative mutant or MSK1 knockdown cells. Thus, MSK1 is required for tumor promoter-induced cell transformation through its phosphorylation of histone H3 at Ser(10) and AP-1 activation. PMID- 18381467 TI - Reliability and validity of a questionnaire to measure colorectal cancer screening behaviors: does mode of survey administration matter? AB - Valid and reliable self-report measures of cancer screening behaviors are important for evaluating efforts to improve adherence to guidelines. We evaluated test-retest reliability and validity of self-report of the fecal occult blood test (FOBT), sigmoidoscopy (SIG), colonoscopy (COL), and barium enema (BE) using the National Cancer Institute colorectal cancer screening (CRCS) questionnaire. A secondary objective was to evaluate reliability and validity by mail, telephone, and face-to-face survey administration modes. Consenting men and women, 51 to 74 years old, receiving care at a multispecialty clinic for at least 5 years who had not been diagnosed with colorectal cancer were stratified by prior CRCS status and randomized to survey mode (n = 857). Within survey mode, respondents were randomized to complete a second survey at 2 weeks, 3 months, or 6 months. Comparing self-report with administrative and medical records, concordance estimates were 0.91 for COL, 0.85 for FOBT, 0.85 for SIG, and 0.92 for BE. Overall sensitivity estimates were 0.91 for COL, 0.82 for FOBT, 0.76 for SIG, and 0.56 for BE. Specificity estimates were 0.91 for COL, 0.86 for FOBT, 0.89 for SIG, and 0.97 for BE. Sensitivity and specificity varied little by survey mode for any test. Report-to-records ratio showed overreporting for SIG (1.1), COL (1.15), and FOBT (1.57), and underreporting for BE (0.82). Reliability at all time intervals was highest for COL; there was no consistent pattern according to survey mode. This study provides evidence to support the use of the National Cancer Institute CRCS questionnaire to assess self-report with any of the three survey modes. PMID- 18381468 TI - Accuracy of self-reported cancer-screening histories: a meta-analysis. AB - BACKGROUND: Survey data used to study trends in cancer screening may overestimate screening utilization while potentially underestimating existing disparities in use. METHODS: We did a literature review and meta-analysis of validation studies examining the accuracy of self-reported cancer-screening histories. We calculated summary random-effects estimates for sensitivity and specificity, separately for mammography, clinical breast exam (CBE), Pap smear, prostate-specific antigen testing (PSA), digital rectal exam, fecal occult blood testing, and colorectal endoscopy. RESULTS: Sensitivity was highest for mammogram, CBE, and Pap smear (0.95, 0.94, and 0.93, respectively) and lowest for PSA and digital rectal exam histories (0.71 and 0.75). Specificity was highest for endoscopy, fecal occult blood testing, and PSA (0.90, 0.78, and 0.73, respectively) and lowest for CBE, Pap smear, and mammogram histories (0.26, 0.48, and 0.61, respectively). Sensitivity and specificity summary estimates tended to be lower in predominantly Black and Hispanic samples compared with predominantly White samples. When estimates of self-report accuracy from this meta-analysis were applied to cancer screening prevalence estimates from the National Health Interview Survey, results suggested that prevalence estimates are artificially increased and disparities in prevalence are artificially decreased by inaccurate self-reports. CONCLUSIONS: National survey data are overestimating cancer-screening utilization for several common procedures and may be masking disparities in screening due to racial/ethnic differences in reporting accuracy. PMID- 18381470 TI - The effects of survey mode and asking about future intentions on self-reports of colorectal cancer screening. AB - Colorectal cancer (CRC) screening rates are often ascertained via self-reports but can be subject to overreporting bias. Asking about intention to get screened before asking about past screening may minimize overreporting of cancer screening. In a statewide survey conducted from July through October of 2005, we embedded an experiment that tested the effect of question ordering (asking about future intention to get screened before or after asking about past screening; "future first" and "future second," respectively), crossed with survey mode (mail versus telephone), on CRC screening rates. Weighted analysis focused on 752 respondents who were ages 50 years or older. We found (a) that asking about future intentions to get screened before asking about past screening (future first) statistically significantly lowers reports of past CRC screening [70.9% future second versus 58.0% future first; odds ratio (OR), 1.83; 95% confidence interval (95% CI), 1.08-3.13]; (b) that there was no main effect of survey mode; and (c) that the effect of the ordering of the future intentions item varies by survey mode. In the mailed survey, the odds of reporting past CRC screening were almost thrice greater in the future second condition compared with the future first condition (72.4% versus 49.0%, respectively; OR, 2.74; 95% CI, 1.22-6.17). In the telephone condition, the odds of reporting were only 28% higher in the future second (69.5%) condition than in the future first condition (63.9%; OR, 1.28; 95% CI, 0.64-2.57). The results suggest that asking about future intentions to get screened before the actual behavior elicits lower, and arguably more truthful reports of CRC screening but mainly in mailed surveys. PMID- 18381469 TI - Validation of self-reported colorectal cancer (CRC) screening in a study of ethnically diverse first-degree relatives of CRC cases. AB - BACKGROUND: Evidence about the accuracy of self-reports of colorectal cancer (CRC) screening is lacking. We conducted a validation protocol in a randomized trial to increase CRC screening among high-risk individuals. METHODS: First degree relatives (n = 1,280) of CRC cases who were due for CRC screening were included in the parent trial. All subjects who completed the follow-up interview (n = 948) were asked to participate in validation activities. Self-reports of receipt of CRC screening during the 12-month study period were verified via physicians. RESULTS: Although 60% (n = 567) verbally agreed, only 171 subjects (18% of original sample) returned the signed validation form with the physician name and contact information and a medical information release statement. The signed forms were mailed to physicians with a $10 incentive and the request to list the dates of recent CRC screening tests. One hundred twenty-three physicians (72% of physicians contacted, 13% of original sample) returned completed validation forms. Rates of agreement were low across all three screening types with physicians verifying self-reported screening for 29% of fecal occult blood testing, 56% of sigmoidoscopy, 55% of colonoscopy, and 57% of any screening test. CONCLUSION: Validation of self-report using the type of protocol we used for subjects receiving medical care in many community settings may be unfeasible and cost inefficient. Given the overall low participation rate in validation activities and considerable challenges in collecting high quality data, conclusions about the accuracy of self-reported CRC screening are difficult to make based on the results of this study. PMID- 18381471 TI - Evaluation of claims, medical records, and self-report for measuring fecal occult blood testing among medicare enrollees in fee for service. AB - BACKGROUND: There is no agreement on the best data source for measuring colorectal cancer (CRC) screening. Medicare claims have been used to measure CRC testing but the validity of using claims to measure fecal occult blood tests (FOBT) has not been established. METHODS: We compared ascertainment of FOBT among three data sources: self-reports, Medicare claims, and medical records. Data were collected on FOBT use during the study window (1/1/1998 - 12/31/2002). Our study was conducted with North Carolina Medicare enrollees (N = 561) who had previously responded to a telephone survey on CRC tests. FOBT information was abstracted from respondents' physician office medical records and compared with self reported FOBT use and Medicare claims for FOBT. Data sources were assessed for accuracy and completeness of FOBT reporting using sensitivity, specificity, positive predictive value, negative predictive value, and agreement. RESULTS: Reporting of FOBT use in the prior year in medical records and Medicare claims agreed 82% of the time [95% confidence interval (95% CI), 79-85%]. FOBT 1-year use rates from self-report agreed with test use found in medical records 70% of the time (95% CI, 66-74%). The lowest agreement was between self-reported 1-year FOBT use and Medicare claims, which agreed 67% of the time (95% CI, 63-71%). CONCLUSIONS: No data source could be established as providing complete and valid information about FOBT use among Medicare enrollees, showing the difficulty of ascertaining test use rates for noninvasive, low-cost procedures conducted in multiple settings. Caution should be used when attempting to measure FOBT use with self-report, Medicare claims, or medical records. PMID- 18381472 TI - Serum levels of vitamin D metabolites and breast cancer risk in the prostate, lung, colorectal, and ovarian cancer screening trial. AB - Experimental and epidemiologic studies suggest that vitamin D metabolites (1,25 dihydroxyvitamin D [1,25(OH)(2)D] and its precursor 25-hydroxyvitamin D [25(OH)D]) may reduce breast cancer risk. We examined subsequent breast cancer risk related to serum levels of these metabolites. In a cohort of women ages 55 to 74 years, who donated blood at baseline (1993-2001) in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial, we identified 1,005 incident breast cancer cases during follow-up through 2005 (mean time between blood draw and diagnosis, 3.9 years). Noncases (n = 1,005) were frequency matched to the cases based on age and year of entry. Sample weights that accounted for unequal probabilities of selecting cases and noncases were applied to make inferences that reflected the entire Prostate, Lung, Colorectal, and Ovarian cohort. Using Cox proportional hazards modeling, we computed breast cancer relative risks (RR) and 95% confidence intervals (95% CI) by quintile for each metabolite. The RR of breast cancer for the highest quintile of 25(OH)D concentration versus the lowest was 1.04 (95% CI, 0.75-1.45; P(trend) = 0.81). Similarly, the breast cancer RR for the highest quintile of 1,25(OH)(2)D compared with the lowest was 1.23 (95% CI, 0.91-1.68; P(trend) = 0.14). Excluding the first 2 years of follow-up did not materially alter these estimates. There was also no evidence of inverse risk in older women (> or =60 years) versus younger women (<60 years). In this prospective study of postmenopausal women, we did not observe an inverse association between circulating 25(OH)D or 1,25(OH)(2)D and breast cancer risk, although we cannot exclude an association in younger women or with long-term or earlier exposure. PMID- 18381473 TI - Height, body mass index, and ovarian cancer: a pooled analysis of 12 cohort studies. AB - BACKGROUND: Although many studies have investigated the association between anthropometry and ovarian cancer risk, results have been inconsistent. METHODS: The associations of height, body mass index (BMI), and ovarian cancer risk were examined in a pooled analysis of primary data from 12 prospective cohort studies from North America and Europe. The study population consisted of 531,583 women among whom 2,036 epithelial ovarian cancer cases were identified. To summarize associations, study-specific relative risks (RR) were estimated using the Cox proportional hazards model and then combined using a random-effects model. RESULTS: Women with height > or =1.70 m had a pooled multivariate RR of 1.38 [95% confidence interval (95% CI), 1.16-1.65] compared with those with height <1.60 m. For the same comparison, multivariate RRs were 1.79 (95% CI, 1.07-3.00) for premenopausal and 1.25 (95% CI, 1.04-1.49) for postmenopausal ovarian cancer (P(interaction) = 0.14). The multivariate RR for women with a BMI > or =30 kg/m(2) was 1.03 (95% CI, 0.86-1.22) compared with women with a BMI from 18.5 to 23 kg/m(2). For the same comparison, multivariate RRs were 1.72 (95% CI, 1.02 2.89) for premenopausal and 1.07 (95% CI, 0.87-1.33) for postmenopausal women (P(interaction) = 0.07). There was no statistically significant heterogeneity between studies with respect to height or BMI. BMI in early adulthood was not associated with ovarian cancer risk. CONCLUSION: Height was associated with an increased ovarian cancer risk, especially in premenopausal women. BMI was not associated with ovarian cancer risk in postmenopausal women but was positively associated with risk in premenopausal women. PMID- 18381474 TI - Validation of self-reported colorectal cancer screening behavior from a mixed mode survey of veterans. AB - OBJECTIVE: The aim of the study was to validate self-reported colorectal cancer (CRC) screening using the National Cancer Institute Colorectal Cancer Screening questionnaire. MATERIALS AND METHODS: 890 patients, ages 50 to 75 years, from the Minneapolis Veterans Affairs (VA) Medical Center were surveyed by mail. Phone administration was attempted with mail nonresponders. VA and non-VA records were combined for the reference standard. Sensitivity, specificity, concordance, and report-to-records ratio (R2R) were estimated for overall and test-specific CRC adherence among respondents providing complete medical records. Secondary analyses examined variation in estimates by patient characteristics, treatment of missing and uncertain responses, and whether a strict or liberal time interval was used for assessing concordance. RESULTS: Complete medical records were available for 345 of the 686 survey responders. For overall adherence, sensitivity was 0.98, specificity was 0.59, concordance was 0.88, and R2R was 1.14. Sensitivity was 0.82 for fecal occult blood test (FOBT), 0.75 for sigmoidoscopy, 0.97 for colonoscopy, and 0.63 for double-contrast barium enema (DCBE). Specificity was 0.89 for FOBT, 0.76 for sigmoidoscopy, 0.72 for colonoscopy, and 0.85 for DCBE. Concordance was >0.80 for all tests other than sigmoidoscopy (0.76). R2R was 1.31 for FOBT, 1.33 for sigmoidoscopy, 1.42 for colonoscopy, and 6.13 for DCBE. The R2R was lower for a combined sigmoidoscopy and colonoscopy measure. Overreporting was more pronounced for older, less educated individuals with no family history of CRC. Sensitivity and R2R improved using a liberal interval and treating uncertain responses as nonadherent (versus missing), but differences were not statistically significant. CONCLUSIONS: Self reported CRC screening validity is generally acceptable and robust across definitional decisions, but varies by screening test and patient characteristics. PMID- 18381475 TI - Prediagnostic use of hormone therapy and mortality after breast cancer. AB - BACKGROUND: A few studies have observed reduced breast cancer mortality in women who used hormone therapy before diagnosis. Due to the high prevalence of past and current hormone use, it is important to investigate whether these preparations are related to breast cancer mortality. METHODS: To evaluate the influence of prediagnostic use of hormone therapy on breast cancer mortality, a prospective cohort of 12,269 women ages 50 years or more diagnosed with incident invasive breast cancer and residents of Wisconsin, Massachusetts, or New Hampshire were enrolled in three phases beginning in 1988. They were followed for death until December 31, 2005, using the National Death Index. Cumulative mortality and multivariable adjusted hazard rate ratios for breast cancer and other mortality causes were calculated for women according to any hormone therapy use, and for exclusive use of estrogen or estrogen-progestin (EP). RESULTS: During an average 10.3 years of follow-up, 1,690 deaths from breast cancer were documented. Cumulative mortality from breast cancer was lower among hormone therapy users, specifically current users at the time of diagnosis, and EP users, compared with nonusers. Adjusted survival varied by type and duration of hormone therapy before diagnosis. A reduced risk of death from breast cancer was associated with EP preparations (hazard rate ratio, 0.73; 0.59-0.91) and with > or =5 years of EP use (0.60; 0.43-0.84). No association was observed for women who were former or current users of E-alone preparations. CONCLUSIONS: Although use of combined EP preparations increases breast cancer risk, in this study, use of these hormones before diagnosis was associated with reduced risk of death after a breast cancer diagnosis. The better survival among users, particularly of EP, persisted after adjustment of screening, stage, and measured confounders. PMID- 18381476 TI - Validity of four self-reported colorectal cancer screening modalities in a general population: differences over time and by intervention assignment. AB - Little is known about the validity of self-reported colorectal cancer screening. To date, few published studies have validated all four screening modalities per recommended guidelines or included a general population-based sample, and none has assessed validity over time and by intervention condition. To estimate the validity of self-reported screening, a random sample of 200 adults, ages > or =50 years, was selected from those completing annual surveys on screening behavior as part of an intervention study. Approximately 60% of the validation sample authorized medical record review. Sensitivity, specificity, and positive and negative predictive values were calculated for baseline and year 1 follow-up reports for each test and for overall screening adherence. Sensitivity at baseline ranged from 86.9% (flexible sigmoidoscopy) to 100% (colonoscopy). Sensitivity at follow-up was slightly lower. Adjusting for validity measures, the sample overreported screening prevalence at baseline for each of the four modalities. At follow-up, overreporting was greatest for fecal occult blood test (13.0%). Overreporting across intervention conditions was highest for fecal occult blood test (10.8% for control; 24.8% for the most intense intervention) and overall screening adherence (10.9% for control; 14.3% for the most intense intervention). Sensitivity and specificity of self-reported colorectal cancer screening compared with medical records were high; however, adjusting self reported screening rates based on relative error rates reduced screening prevalence estimates. Those exposed to more intense interventions to modify screening behavior seemed more likely to overestimate their screening rates compared with those who were not exposed. PMID- 18381477 TI - Intended coping responses to cancer symptoms in healthy adults: the roles of symptom knowledge, detection behavior, and perceived threat. AB - BACKGROUND: To date, the causal effects of the knowledge of cancer-related symptoms and detection behavior on coping with cancer-related symptoms have not been identified. Therefore, the current study explored the effects of active or passive detection of supposedly well-known or less-known cancer-related symptoms on intended coping responses. In addition, we were interested in the extent to which these effects are driven by heightened perceptions of threat. METHODS: In an experimental study using a 2 x 2 within-subject design, 221 Dutch adults from the general population responded to a survey study sent to their homes (18.4% response). They were asked to read scenario information about four cancer-related symptoms that were (a) well known or less known and (b) actively or passively detected (e.g., self-examination versus unusual blood loss). The authors measured intended coping responses to the detection of cancer-related symptoms as either adaptive (e.g., visiting a general practitioner) or maladaptive (e.g., denial of the symptom). RESULTS: As expected, the findings revealed that well-known symptoms resulted in more anticipated adaptive coping and less anticipated maladaptive coping than less-known symptoms. Unfortunately, the findings also suggest that the active as opposed to passive detection of cancer symptoms (e.g., self-examination versus unusual blood loss) is likely to result in more maladaptive coping. These effects were mediated by heightened perceptions of threat. CONCLUSIONS: Future health education programs that aim to motivate people to be more active in the early detection of cancer symptoms should first focus on increasing people's knowledge about the early warning signs of cancer. PMID- 18381478 TI - Food-induced anaphylaxis and oral allergy syndrome. PMID- 18381479 TI - Hepatitis B in children. PMID- 18381480 TI - Bleeding disorders. PMID- 18381481 TI - Index of suspicion. PMID- 18381482 TI - Pediatrics in the community: a smoking ban in the heart of tobacco country. PMID- 18381483 TI - Tuberculosis. PMID- 18381484 TI - Wilms tumor. PMID- 18381486 TI - Can home AEDs improve survival? PMID- 18381487 TI - Pharmacokinetic-pharmacodynamic modeling of biomarker response and tumor growth inhibition to an orally available cMet kinase inhibitor in human tumor xenograft mouse models. AB - (R)-3-[1-(2,6-Dichloro-3-fluoro-phenyl)-ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-4 yl)-pyridin-2-ylamine (PF02341066) was identified as an orally available, ATP competitive small molecule inhibitor of cMet receptor tyrosine kinase. The objectives of the present studies were to characterize 1) the pharmacokinetic pharmacodynamic relationship of the plasma concentrations of PF02341066 to cMet phosphorylation in tumor (biomarker) and 2) the relationship of cMet phosphorylation to antitumor efficacy (pharmacological response). Athymic mice implanted with GTL16 gastric carcinoma or U87MG glioblastoma xenografts were treated with PF02341066 once daily at doses selected to encompass ED(50) values. Plasma concentrations of PF02341066 were best described by a one-compartment pharmacokinetic model. A time-delay (hysteresis) was observed between the plasma concentrations of PF02341066 and the cMet phosphorylation response. A link model was therefore used to account for this hysteresis. The model fitted the time courses of cMet phosphorylation well, suggesting that the main reason for the hysteresis is a rate-limiting distribution from plasma into tumor. The EC(50) and EC(90) values were estimated to be 19 and 167 ng/ml, respectively. For tumor growth inhibition, the exponential tumor growth model fitted the time courses of individual tumor growth inhibition well. The EC(50) for the GTL16 tumor growth inhibition was estimated to be 213 ng/ml. Thus, the EC(90) for the inhibition of cMet phosphorylation corresponded to the EC(50) for the tumor growth inhibition, suggesting that near-complete inhibition of cMet phosphorylation (>90%) is required to significantly inhibit tumor growth (>50%). The present results will be helpful in determining the appropriate dosing regimen and in guiding dose escalation to rapidly achieve efficacious systemic exposure in the clinic. PMID- 18381488 TI - Prediction of pharmacokinetic drug-drug interactions using human hepatocyte suspension in plasma and cytochrome P450 phenotypic data. III. In vitro-in vivo correlation with fluconazole. AB - Whereas ketoconazole is often used to study the worst-case scenario for clinical pharmacokinetic drug-drug interactions (DDIs) for drugs that are primarily metabolized by CYP3A4, fluconazole is considered to be a moderate inhibitor of CYP3A4, providing assessment of the moderate-case scenario of CYP3A-based DDIs. Fluconazole is also a moderate inhibitor of CYP2C9 and CYP2C19. For predicting clinical DDIs using conventional approaches, determining the in vivo inhibitor concentration at the enzymatic site [I], a critical parameter, is still not practical. In our previous study, a novel method involving hepatocyte suspension in plasma was used to circumvent the need to determine the elusive [I] value. In this study, the CYP1A2, 2C9, 2C19, 2D6, and 3A4 activities remaining in the presence of fluconazole were determined in human hepatocytes suspended in human plasma, covering a range of fluconazole clinical plasma concentrations (C(avg) and C(max)). Because the protein-binding effect of fluconazole is expected to be close to that in vivo, the inhibition observed in vitro will be similar to that in vivo. This inhibition information was then applied to the cytochrome P450 (P450) phenotypic data to predict DDIs. Using the available P450 phenotypic information on theophylline, tolbutamide, omeprazole, S-warfarin, phenytoin, cyclosporine, and midazolam and that determined in this study for sirolimus and tacrolimus, we found that the predictions for area under the curve increases for most of these drugs in the presence of fluconazole were remarkably similar (within 35%) to the observed clinical values. This study proves the general applicability of our approach using human hepatocyte incubation in human plasma to predict DDIs. PMID- 18381489 TI - Prediction of pharmacokinetic drug-drug interactions using human hepatocyte suspension in plasma and cytochrome P450 phenotypic data. II. In vitro-in vivo correlation with ketoconazole. AB - Traditional cytochrome P450 (P450) based drug-drug interaction (DDI) predictions are based on the ratio of an inhibitor's physiological concentration [I] and its inhibition constant K(i). Determining [I] at the enzymatic site, although critical for predicting clinical DDIs, remains a technical challenge. In our previous study, a novel approach using cryopreserved human hepatocytes suspended in human plasma was investigated to mimic the in vivo concentration of ketoconazole at the enzymatic site (Lu et al., 2007), effectively eliminating the estimation of the elusive [I] value. P450 inhibition in this system appears to model that in vivo. Using the ketoconazole inhibition information in a human hepatocyte-plasma suspension together with quantitative P450 phenotypic information, we successfully predicted the pharmacokinetic DDIs for a small set of drugs, such as theophylline, tolbutamide, omeprazole, desipramine, midazolam, loratadine, cyclosporine, and alprazolam, as well as an investigational compound. For the applicability of this model on a wider scale the in vitro-in vivo correlation data set needed to be expanded. However, for most drugs in the literature there is not enough quantitative information on the involvement of individual P450s to predict DDIs retrospectively. To facilitate that, in this study we determined quantitative P450 phenotyping for seven marketed drugs: budesonide, buprenorphine, loratadine, sirolimus, tacrolimus, docetaxel, and methylprednisolone. Augmentation of the new data set with the one generated previously produced broader a database that provided further support for the wider applicability of this approach using ketoconazole as a potent CYP3A inhibitor. This application is predicted to be equally effective with other P450 inhibitors that are not substrates of efflux pumps. PMID- 18381490 TI - Underage drinking: understanding and reducing risk in the context of human development. Foreword. PMID- 18381491 TI - Underage drinking: understanding and reducing risk in the context of human development. Editors' preface. PMID- 18381492 TI - Underage drinking: a developmental framework. AB - A developmental framework for understanding and addressing the problem of underage alcohol consumption is presented. The first section presents the rationale for a developmental approach, including striking age-related data on patterns of onset, prevalence, and course of alcohol use and disorders in young people. The second section examines the fundamental meaning of a developmental approach to conceptualizing underage drinking. The third section delineates contemporary principles of developmental psychopathology as a guide to future research and intervention efforts. Strategic, sensitive, and effective efforts to address the problem of underage drinking will require a developmentally informed approach to research, prevention, and treatment. PMID- 18381493 TI - Early developmental processes and the continuity of risk for underage drinking and problem drinking. AB - Developmental pathways to underage drinking emerge before the second decade of life. Many scientists, however, as well as the general public, continue to focus on proximal influences surrounding the initiation of drinking in adolescence, such as social, behavioral, and genetic variables related to availability and ease of acquisition of the drug, social reinforcement for its use, and individual differences in drug responses. In the past 20 years, a considerable body of evidence has accumulated on the early (often much earlier than the time of the first drink) predictors and pathways of youthful alcohol use and abuse. These early developmental influences involve numerous risk, vulnerability, promotive, and protective processes. Some of these factors are not related directly to alcohol use, whereas others involve learning and expectancies about later drug use that are shaped by social experience. The salience of these factors (identifiable in early childhood) for understanding the course and development of adult alcohol and other drug use disorders is evident from the large and growing body of findings on their ability to predict adult clinical outcomes. This review summarizes the evidence on early pathways toward and away from underage drinking, with a particular focus on the risk and protective factors and the mediators and moderators of risk for underage drinking that become evident during the preschool and early school years. It is guided by a developmental perspective on the aggregation of risk and protection and examines the contributions of biological, psychological, and social processes within the context of normal development. Implications of this evidence for policy, intervention, and future research are discussed. PMID- 18381485 TI - Home use of automated external defibrillators for sudden cardiac arrest. AB - BACKGROUND: The most common location of out-of-hospital sudden cardiac arrest is the home, a situation in which emergency medical services are challenged to provide timely care. Consequently, home use of an automated external defibrillator (AED) might offer an opportunity to improve survival for patients at risk. METHODS: We randomly assigned 7001 patients with previous anterior-wall myocardial infarction who were not candidates for an implantable cardioverter defibrillator to receive one of two responses to sudden cardiac arrest occurring at home: either the control response (calling emergency medical services and performing cardiopulmonary resuscitation [CPR]) or the use of an AED, followed by calling emergency medical services and performing CPR. The primary outcome was death from any cause. RESULTS: The median age of the patients was 62 years; 17% were women. The median follow-up was 37.3 months. Overall, 450 patients died: 228 of 3506 patients (6.5%) in the control group and 222 of 3495 patients (6.4%) in the AED group (hazard ratio, 0.97; 95% confidence interval, 0.81 to 1.17; P=0.77). Mortality did not differ significantly in major prespecified subgroups. Only 160 deaths (35.6%) were considered to be from sudden cardiac arrest from tachyarrhythmia. Of these deaths, 117 occurred at home; 58 at-home events were witnessed. AEDs were used in 32 patients. Of these patients, 14 received an appropriate shock, and 4 survived to hospital discharge. There were no documented inappropriate shocks. CONCLUSIONS: For survivors of anterior-wall myocardial infarction who were not candidates for implantation of a cardioverter defibrillator, access to a home AED did not significantly improve overall survival, as compared with reliance on conventional resuscitation methods. (ClinicalTrials.gov number, NCT00047411 [ClinicalTrials.gov].). PMID- 18381494 TI - Transitions into underage and problem drinking: developmental processes and mechanisms between 10 and 15 years of age. AB - Numerous developmental changes occur across levels of personal organization (eg, changes related to puberty, brain and cognitive-affective structures and functions, and family and peer relationships) in the age period of 10 to 15 years. Furthermore, the onset and escalation of alcohol use commonly occur during this period. This article uses both animal and human studies to characterize these multilevel developmental changes. The timing of and variations in developmental changes are related to individual differences in alcohol use. It is proposed that this integrated developmental perspective serve as the foundation for subsequent efforts to prevent and to treat the causes, problems, and consequences of alcohol consumption. PMID- 18381497 TI - Developmentally informed research on the effectiveness of clinical trials: a primer for assessing how developmental issues may influence treatment responses among adolescents with alcohol use problems. AB - The goal of this article is to familiarize readers with the adolescent developmental issues and processes most likely to affect responses to treatment for alcohol use problems. Although the need for research that blends developmental science and treatment outcome research is widely acknowledged, scant information exists about developmentally informed approaches to treatment research with alcohol-abusing teens. Exactly how developmental issues may influence treatment responses among adolescents with alcohol use problems remains an open question. In the hope of moving developmentally informed research forward, this article reports findings from a literature review regarding the degree to which developmental issues and processes have been considered in adolescent alcohol treatment research. Moreover, promising concepts and methods from applied developmental science are discussed, as are various developmental processes and transitions that may influence adolescent risk behavior. Finally, guidance is provided regarding how applied developmental science conceptualizations and methods may be incorporated successfully into randomized, clinical trials with adolescents with alcohol use problems. PMID- 18381495 TI - A developmental perspective on alcohol and youths 16 to 20 years of age. AB - Late adolescence (ie, 16-20 years of age) is a period characterized by escalation of drinking and alcohol use problems for many and by the onset of an alcohol use disorder for some. This heightened period of vulnerability is a joint consequence of the continuity of risk from earlier developmental stages and the unique neurologic, cognitive, and social changes that occur in late adolescence. We review the normative neurologic, cognitive, and social changes that typically occur in late adolescence, and we discuss the evidence for the impact of these transitions on individual drinking trajectories. We also describe evidence linking alcohol abuse in late adolescence with neurologic damage and social impairments, and we discuss whether these are the bases for the association of adolescent drinking with increased risks of mental health, substance abuse, and social problems in adulthood. Finally, we discuss both the challenges and successes in the treatment and prevention of adolescent drinking problems. PMID- 18381498 TI - Evidence-based treatments for alcohol use disorders in adolescents. AB - The prevalence of adolescent alcohol use and its related consequences underscore the need for evidenced-based treatments in this population. During the past decade, much progress has been made in treating adolescent alcohol use disorders with evidenced-based modalities developed specifically for adolescents. Controlled treatment outcome studies that compared > or = 1 modality, used random assignment to treatment conditions, and were published between 1990 and 2004 are discussed in this review. Psychosocial treatments such as family-based interventions, motivational enhancement therapy (motivational interviewing), behavioral therapy, and cognitive-behavioral therapy, as well as the limited pharmacotherapy studies, are discussed. All of the studies used assessment tools validated for use in adolescent populations. Overall, great strides have been made in the area of adolescent alcohol treatment, and the treatment modalities presented have more than adequate potential for replication. PMID- 18381496 TI - Preventive interventions addressing underage drinking: state of the evidence and steps toward public health impact. AB - The epidemiological features of underage drinking and evidence of its social, health, and economic consequences suggest compelling reasons for the development and dissemination of effective preventive interventions. To clarify the nature and extent of the current evidence base on preventive interventions addressing underage drinking, a review of the literature was conducted through extensive searches of the research literature on outcome evaluations, existing reviews of this body of outcome research (N = 25), and summary reports of evidence on specific interventions. More than 400 interventions were identified and screened, and the evidence for 127 was reviewed. Criteria for the evaluation of evidence were established for intervention studies with alcohol-specific outcome measures for 3 developmental periods (< 10, 10-15, and 16 to > or = 20 years of age). Ultimately, 12 interventions met criteria for "most promising" evidence and 29 met criteria for "mixed or emerging" evidence. Conducting this review revealed clear advances in the number of evidence-based interventions available and the quality of outcome research; however, much work remains to achieve greater public health impact through evidence-based interventions. This work should consider (1) the great need for intervention research related to understudied developmental phases, intervention domains (eg, family, school, community, and media), and populations (eg, early tweens, late teens, young adults not attending college, and nonmajority populations); (2) the critical importance of addressing key issues in research design and methods (eg, limited longitudinal studies, replication studies, and dissemination research); and (3) the need for improved consistency in application of evidence and reporting standards. Finally, we recommend the application of emerging consumer-oriented and community participatory models for intervention development and research, designed to increase the likelihood of "real-world" public health impact through improved translation of intervention science into practice. PMID- 18381499 TI - School readiness. AB - School readiness includes the readiness of the individual child, the school's readiness for children, and the ability of the family and community to support optimal early child development. It is the responsibility of schools to be ready for all children at all levels of readiness. Children's readiness for kindergarten should become an outcome measure for community-based programs, rather than an exclusion criterion at the beginning of the formal educational experience. Our new knowledge of early brain and child development has revealed that modifiable factors in a child's early experience can greatly affect that child's learning trajectory. Many US children enter kindergarten with limitations in their social, emotional, cognitive, and physical development that might have been significantly diminished or eliminated through early identification of and attention to child and family needs. Pediatricians have a role in promoting school readiness for all children, beginning at birth, through their practices and advocacy. The American Academy of Pediatrics affords pediatricians many opportunities to promote the physical, social-emotional, and educational health of young children, with other advocacy groups. This technical report supports American Academy of Pediatrics policy statements "Quality Early Education and Child Care From Birth to Kindergarten" and "The Inappropriate Use of School 'Readiness' Tests." PMID- 18381500 TI - Prevention of influenza: recommendations for influenza immunization of children, 2007-2008. AB - The American Academy of Pediatrics recommends annual influenza immunization for all children with high-risk conditions who are 6 months of age and older, for all healthy children ages 6 through 59 months, for all household contacts and out-of home caregivers of children with high-risk conditions and of healthy children younger than 5 years, and for all health care professionals. To more fully protect against the morbidity and mortality of influenza, increased efforts are needed to identify and immunize all children at high risk and all healthy children ages 6 through 59 months and to inform their parents when annual immunizations are due. Previously unimmunized children who are at least 6 months of age but younger than 9 years should receive 2 doses of influenza vaccine, given 1 month apart, beginning as soon as possible on the basis of local availability during the influenza season. If children in this cohort received only 1 dose for the first time in the previous season, it is recommended that 2 doses be administered in the current season. This recommendation applies only to the influenza season that follows the first year that a child younger than 9 years receives influenza vaccine. A child who then also fails to receive 2 doses the next year should be given only 1 dose per year from that point on. Influenza vaccine should also continue to be offered throughout the influenza season, even after influenza activity has been documented in a community. On the basis of global surveillance of circulating virus strains, the influenza vaccine may change from year to year; indeed, 1 of the 3 strains in the 2007-2008 vaccine is different from the previous year's vaccine. All health care professionals, influenza campaign organizers, and public health agencies should develop plans for expanding outreach and infrastructure to immunize all children for whom influenza vaccine is recommended. Appropriate prioritization of administering influenza vaccine will also be necessary when vaccine supplies are delayed or limited. Because the influenza season often extends into March, immunization against influenza is recommended to continue through late winter and early spring. Lastly, it is recommended that for the 2007-2008 season, and likely beyond, health care professionals do not prescribe amantadine or rimantadine for influenza treatment or chemoprophylaxis, because widespread resistance to these antiviral medications now exists among influenza A viral strains. However, oseltamivir and zanamivir can be prescribed for treatment or chemoprophylaxis, because influenza A and B strains remain susceptible. PMID- 18381501 TI - Impact of admission-day crowding on the length of stay of pediatric hospitalizations. AB - OBJECTIVE: Increased crowding may affect the care that is delivered to hospitalized patients, particularly around the time of admission. There is little information about the impact of admission-day crowding on the outcome of children who are hospitalized with common pediatric conditions. METHODS: A population cohort was constructed of children who were aged 1 to 17 years and were hospitalized in Pennsylvania and New York between April 1, 1996, and June 30, 1998, with 1 of 19 common pediatric conditions (N = 116,235). Condition-specific Cox regression and logit models were developed to estimate the effect of admission-day occupancy on 4 outcome measures after controlling for illness severity and site of care: length of stay; 21-day readmission; prolonged stay or a stay longer than the typical, uncomplicated stay for that condition as a measure of care delivered to patients with uncomplicated courses; and conditional length of stay as a measure of care delivered to patients whose stays are prolonged. RESULTS: For children who were admitted with respiratory disease, increasing admission-day occupancy from 60% to 100% was associated with a 0.25 day increase in the average length of stay. Increased admission-day occupancy above 60% was also associated with higher odds of a prolonged stay but not with a change in 21-day readmission rates or conditional length of stay. For children who were admitted with nonrespiratory conditions, increased admission-day occupancy was not associated with changes to any length-of-stay outcome. CONCLUSIONS: Increased admission-day occupancy was associated with longer lengths of stay for less complicated respiratory admissions but not for children who were admitted with the most serious conditions. These results suggest that medical professionals, during times of increased workload, first focus their attention on more acutely ill children with a complicated course and thus delay treatment of children who have less complicated courses but require time-consuming management and treatment. PMID- 18381502 TI - Effect of a pediatric practice-based smoking prevention and cessation intervention for adolescents: a randomized, controlled trial. AB - OBJECTIVE: The purpose of this work was to determine whether a pediatric practice based smoking prevention and cessation intervention increases abstinence rates among adolescents. METHODS: Eight pediatric primary care clinics were randomly assigned to either intervention or usual care control condition. The provider- and peer-delivered intervention tested was based on the 5A model recommended by the US Public Health Service clinical practice guidelines and the American Academy of Pediatrics and consisted of brief counseling by the pediatric provider followed by 1 visit and 4 telephone calls by older peer counselors aged 21 to 25 years. A consecutive sample of patients aged 13 to 17 years scheduled for an office visit was eligible regardless of smoking status. Of 2711 patients who agreed to participate, 2709 completed baseline assessments, and 2700 (99.6%) and 2690 (99.2%) completed 6- and 12-month assessments, respectively. RESULTS: Compared with the usual care condition, nonsmokers who received the provider- and peer-delivered intervention were significantly more likely to self-report having remained abstinent at 6-month and 12-month follow-up; smokers who received the provider- and peer-delivered intervention were more likely to report having quit at the 6-month but not the 12-month follow-up. A number of adolescent characteristics (eg, age, peer smoking, tobacco dependence, and susceptibility) were found to be predictive of abstinence at follow-up. CONCLUSIONS: A pediatric practice-based intervention delivered by pediatric providers and older peer counselors proved feasible and effective in discouraging the initiation of smoking among nonsmoking adolescents for 1 year and in increasing abstinence rates among smokers for 6 months. PMID- 18381503 TI - Inattention, hyperactivity, and school performance in a population of school-age children with complex congenital heart disease. AB - INTRODUCTION: There is a growing interest in characterizing the neurodevelopmental outcomes of school-age survivors of cardiac surgery. The purpose of this study was to examine a population of 5- to 10-year-old children who underwent newborn cardiac surgery for complex congenital heart disease to characterize and assess risk factors for problems with inattention and hyperactivity, as well as the use of remedial school services. PATIENTS AND METHODS: This study was a cross-sectional analysis of patients who underwent newborn cardiac surgery and were enrolled in a neuroprotection trial conducted at our institution between 1992 and 1997. Parents and teachers completed questionnaires for the school-age child to elicit information pertaining to the child's general health and academic performance. The severity of hyperactivity and inattention were assessed by using 2 standardized questionnaires (Attention Deficit/Hyperactivity Disorder Rating Scale-IV and Behavior Assessment System for Children). In addition to calculating descriptive estimates of their occurrence, single-covariate logistic regression models were specified and tested by using 3 different outcomes (inattention, hyperactivity, and use of remedial school services) and 14 different covariates representing preoperative, intraoperative, and postoperative factors. RESULTS: Data were obtained from parents and/or teachers for 109 children. Fifty-three (49%) were receiving some form of remedial academic services, and 15% were assigned to a special-education classroom. The number of children receiving clinically significant scores for inattention and hyperactivity on the Behavior Assessment System for Children was 3 to 4 times higher than observed in the general population. On the Attention Deficit/Hyperactivity Disorder Rating Scale-IV, 30% of the parents reported high risk scores for inattention and 29% reported high-risk scores for hyperactivity. No perioperative factors were statistically associated with adverse outcomes. CONCLUSION: In this cohort of children with complex congenital heart disease, a significant proportion of the children were at risk for inattention and hyperactivity, and nearly half were using remedial school services. We did not identify any perioperative risk factors, which correlated with high-risk scores or the use of remedial school services. Ongoing neurodevelopmental follow-up and screening are recommended in this vulnerable population. PMID- 18381504 TI - Vaccination coverage by special health care needs status in young children. AB - OBJECTIVE: The purpose of this study was to compare vaccination coverage among children 19 to 35 months of age with and without special health care needs overall and among demographic subgroups. METHODS: Data are from the National Survey of Children With Special Health Care Needs, a module of the State and Local Area Integrated Telephone Survey, which was sponsored by the Health Resources and Services Administration Maternal and Child Health Bureau and conducted in 2000-2002 by the Centers for Disease Control and Prevention National Center for Health Statistics. We used data from the National Immunization Survey and the National Survey of Children With Special Health Care Needs to examine immunization coverage rates for individual vaccines and an array of combined series vaccinations. The relationship between special needs and immunization status was analyzed by age, gender, and race or ethnicity of the child; the child's health insurance type; the mother's educational attainment and presence in the household; and household income relative to the federal poverty level. RESULTS: Overall, there were no significant differences between children with and without special needs for any of the individual antigens or combined immunization series. Some significant differences by special needs status were found within certain demographic subgroups. CONCLUSIONS: Our results suggest that, generally children with special health care needs have immunization rates that are very similar to typically developing children. There is some evidence that children with special health care needs are at risk for underimmunization if they are non Hispanic white or live in an affluent household and are more likely to be immunized if they are Hispanic, poor, publicly insured, or if their mothers did not complete high school. These findings may be due to outreach or support programs that target disadvantaged children. However, it is important to note that the majority of comparisons within demographic subgroups show no significant differences between children with special health care needs and children without special health care needs. PMID- 18381505 TI - Dietary supplements: inpatient policies in US children's hospitals. AB - BACKGROUND: Variations in hospital policies and practices regarding the use of vitamins, minerals, herbs, and other dietary supplements could compromise patient safety and quality of care. We undertook this study to describe pediatric hospital policies regarding dietary supplements. METHODS: We administered a cross sectional survey of a sample of National Association of Children Hospitals and Related Institutions members. Pharmacy personnel were asked about the following: written policies addressing dietary supplements; how such policies are made; and responsibilities for storage, administration, documentation, and checking for potential interactions. We created a 10-point hospital policy and practices quality score to assess the quality of each policy. RESULTS: Of the 186 eligible National Association of Children Hospitals and Related Institutions hospitals, 109 responded (59%). Forty-four percent reported having written policies on vitamins and minerals, herbs, and other dietary supplements. Few hospitals had herbs (2%) or other dietary supplements (38%) on formulary. However, most (81%) allowed patients to use their home supply of dietary supplements under defined circumstances, and 64% allowed staff to make recommendations about dietary supplement use. In 70% of hospitals, nurses stored and administered the home supply of dietary supplements. Less than half (46%) required documentation of a check for drug or dietary supplements interactions. One third (32%) had surgical preoperative recommendations regarding dietary supplements. Only 11% of 109 hospitals satisfied the 10 criteria necessary for a perfect hospital policy and practices quality score indicative of quality practices regarding patient use of dietary supplements. CONCLUSION: Even among those hospitals that have written policies about dietary supplements, there is a wide range in policy quality, which may compromise the safety of patients. PMID- 18381506 TI - Toxicity of buprenorphine overdoses in children. AB - OBJECTIVE: There are few reports in children of overdoses of buprenorphine, a partial opioid agonist used in the treatment of opioid dependence and pain. The purpose of this study was to analyze buprenorphine overdoses in young children reported by US poison centers to the Researched Abuse, Diversion, and Addiction Related Surveillance System. METHODS: A retrospective review of buprenorphine overdoses in children < 6 years of age reported to the Researched Abuse, Diversion, and Addiction-Related Surveillance System from November 2002 through December 2005 was performed. Patients lost to follow-up and those ingesting multiple substances were excluded. RESULTS: Eighty-six cases met inclusion criteria. In the 54 children who developed toxicity, the clinical effects included drowsiness or lethargy (55%), vomiting (21%), miosis (21%), respiratory depression (7%), agitation or irritability (5%), pallor (3%), and coma (2%). There were no fatalities. The mean time to onset of effects was 64.2 minutes, with a range of 20 minutes to 3 hours. Duration of clinical effects was under 2 hours in 11%, 2 to 8 hours in 59%, 8 to 24 hours in 26%, and > 24 hours in 4%. Children who ingested > or = 2 mg of buprenorphine were more likely to experience clinical effects, and all of the children who ingested > 4 mg experienced some effect. No child ingesting < 4 mg experienced a severe effect. Of the 22 children administered naloxone, 67% had at least a partial response. CONCLUSIONS: Buprenorphine overdoses are generally well tolerated in children, with significant central nervous system and respiratory depression occurring in only 7%. Any child ingesting > 2 mg and children < 2 years of age ingesting more than a lick or taste should be referred to the emergency department for a minimum of 6 hours of observation. Naloxone can be used to reverse respiratory depression. PMID- 18381507 TI - Association between living with children and adherence to highly active antiretroviral therapy in the Women's Interagency HIV Study. AB - OBJECTIVE: The purpose of this work was to evaluate whether living with children adversely affects adherence to highly active antiretroviral therapy in HIV infected women. PARTICIPANTS AND METHODS: We conducted a prospective cohort study between October 1998 and September 2005. The study outcome was > or = 95% adherence to highly active antiretroviral therapy evaluated at 5832 semiannual visits among 1366 HIV-infected women in the Women's Interagency HIV Study. The primary exposure defined at the visit immediately before outcome ascertainment was the number of children < or = 18 years of age reported living in the household. RESULTS: The percentage of women who reported > or = 2 children in the household who also reported > or = 95% adherence ranged from 68% to 75% compared with adherence when either 1 child or no children were reported. Each additional child reported living in the household was associated with a 6% decrease in the odds of > or = 95% adherence. CONCLUSION: The impact of living with a child on the ability to take medications by HIV-infected women has not been examined thoroughly. Our data suggest that adherence to highly active antiretroviral therapy is inversely associated with the number of children living in the household. PMID- 18381508 TI - A policy-based school intervention to prevent overweight and obesity. AB - BACKGROUND: The prevalence and seriousness of childhood obesity has prompted calls for broad public health solutions that reach beyond clinic settings. Schools are ideal settings for population-based interventions to address obesity. OBJECTIVE: The purpose of this work was to examine the effects of a multicomponent, School Nutrition Policy Initiative on the prevention of overweight (85.0th to 94.9th percentile) and obesity (> 95.0th percentile) among children in grades 4 through 6 over a 2-year period. METHODS: Participants were 1349 students in grades 4 through 6 from 10 schools in a US city in the Mid Atlantic region with > or = 50% of students eligible for free or reduced-price meals. Schools were matched on school size and type of food service and randomly assigned to intervention or control. Students were assessed at baseline and again after 2 years. The School Nutrition Policy Initiative included the following components: school self-assessment, nutrition education, nutrition policy, social marketing, and parent outreach. RESULTS: The incidences of overweight and obesity after 2 years were primary outcomes. The prevalence and remission of overweight and obesity, BMI z score, total energy and fat intake, fruit and vegetable consumption, body dissatisfaction, and hours of activity and inactivity were secondary outcomes. The intervention resulted in a 50% reduction in the incidence of overweight. Significantly fewer children in the intervention schools (7.5%) than in the control schools (14.9%) became overweight after 2 years. The prevalence of overweight was lower in the intervention schools. No differences were observed in the incidence or prevalence of obesity or in the remission of overweight or obesity at 2 years. CONCLUSION: A multicomponent school-based intervention can be effective in preventing the development of overweight among children in grades 4 through 6 in urban public schools with a high proportion of children eligible for free and reduced-priced school meals. PMID- 18381509 TI - Outcome and growth of infants fetally exposed to heart block-associated maternal anti-Ro52/SSA autoantibodies. AB - OBJECTIVE: The purpose of this work was to analyze outcome with focus on growth in infants fetally exposed to heart block-associated maternal anti-Ro52/SSA autoantibodies and identify maternal factors other than the autoantibodies increasing the risk of fetal heart block. PATIENTS AND METHODS: Thirty-two pregnancies in 30 anti-Ro52-positive mothers were included. Seven fetuses developed second-degree or third-degree atrioventricular block, 8 developed first degree atrioventricular block, and 17 had normal atrioventricular conduction, as diagnosed by using Doppler echocardiography. Three of 6 surviving fetuses with second-degree or third-degree atrioventricular block received treatment with fluorinated steroids. Two fetuses with second-degree atrioventricular block converted to first-degree atrioventricular block without any signs of progression during the study period. Maternal and longitudinal infant data were collected from planned neonatal follow-up and childhood health records from birth to 12 months of age in 31 survivors. RESULTS: Women giving birth to infants with prenatal second-degree or third-degree atrioventricular block were older and with higher parity than those with first-degree atrioventricular block or normal atrioventricular conduction. Second-degree or third-degree atrioventricular block pregnancies were <40 completed weeks, whereas pregnancies with first-degree atrioventricular block or normal atrioventricular conduction had a normal duration. Fetuses with second-degree or third-degree atrioventricular block were retarded by -0.98 +/- 0.77 SD in weight at birth and did not show any catch-up during infancy. In contrast, fetuses with first-degree atrioventricular block or normal atrioventricular conduction had a weight reduction of -0.51 +/- 1.01 SD with a catch-up during the first months after birth. CONCLUSIONS: This report documents that newborns with autoantibody-mediated second-degree or third-degree atrioventricular block are retarded in growth, with no catch-up during infancy, whereas fetuses with first-degree atrioventricular block or normal atrioventricular conduction have a normal growth soon after birth. Increased maternal age and/or parity seem to carry an increased risk for fetal heart block. PMID- 18381510 TI - Maternal smoking and congenital heart defects. AB - OBJECTIVES: In a population-based case-control study, we investigated the association between congenital heart defects and maternal smoking. METHODS: The National Birth Defects Prevention Study enrolled 3067 infants with nonsyndromic congenital heart defects and their parents and 3947 infants without birth defects and their parents. Affected infants had > or = 1 of the following defects: conotruncal, septal, anomalous pulmonary venous return, atrioventricular septal defects, and left-sided or right-sided obstructive heart defects. Mothers of case and control infants were asked if they smoked during the periconceptional period, defined as 1 month before pregnancy through the first trimester. Maternal home and workplace exposure to tobacco smoke during the same period was also determined. Logistic regression was used to compute odds ratios and 95% confidence intervals while controlling for potential confounders. RESULTS: Case infants were more likely to be premature and have lower birth weight than control infants. Women who smoked anytime during the month before pregnancy to the end of the first trimester were more likely to have infants with septal heart defects than women who did not smoke during this time period. This association was stronger for mothers who reported heavier smoking during this period. This relation was independent of potential confounding factors, including prenatal vitamin use, alcohol intake, maternal age, and race or ethnicity. Women who smoked > or = 25 cigarettes per day were more likely than nonsmoking mothers to have infants with right-sided obstructive defects. There was no increased risk of congenital heart defects with maternal exposure to environmental tobacco smoke. CONCLUSIONS: Maternal smoking during pregnancy was associated with septal and right-sided obstructive defects. Additional investigation into the timing of tobacco exposure and genetic susceptibilities that could modify this risk will provide a more precise evidence base on which to build clinical and public health primary prevention strategies. PMID- 18381511 TI - Association of childhood autism spectrum disorders and loss of family income. AB - BACKGROUND: Parents of children with autism have significant out-of-pocket expenditures related to their child's care. The impact of having a child with autism on household income is not known. OBJECTIVE: The purpose of this work was to estimate the loss of household income associated with childhood autism using a nationally representative sample. METHODS: Parents of 11,684 children enrolled in kindergarten to eighth grade were surveyed by the National Household Education Survey-After School Programs and Activities in 2005. An autism spectrum disorder was defined as an affirmative response to the questions, "has a health professional told you that [child] has any of the following disabilities? 1) autism? 2) pervasive developmental disorder or PDD?" There were 131 children with autism spectrum disorder in the sample and 2775 children with other disabilities. We used ordinal logistic regression analyses to estimate the expected income of families of children with autism given their education level and demographic characteristics and compared the expected income with their reported income. RESULTS. Both having a child with autism spectrum disorder and having a child with other disabilities were associated with decreased odds of living in a higher income household after controlling for parental education, type of family, parental age, location of the household, and minority ethnicity. The average loss of annual income associated with having a child with autism spectrum disorder was $6200 or 14% of their reported income. CONCLUSION: Childhood autism is associated with a substantial loss of annual household income. This likely places a significant burden on families in the face of additional out-of-pocket expenditures. PMID- 18381512 TI - Media coverage of the measles-mumps-rubella vaccine and autism controversy and its relationship to MMR immunization rates in the United States. AB - OBJECTIVE: The purpose of this work was to assess the association between media coverage of the MMR-autism controversy and MMR immunization in the United States. METHODS: The public-use files of the National Immunization Survey were used to estimate annual MMR coverage from 1995 to 2004. The primary outcome was selective measles-mumps-rubella nonreceipt, that is, those children who received all childhood immunizations except MMR. Media coverage was measured by using LexisNexis, a comprehensive database of national and local news media. Factors associated with MMR nonreceipt were identified by using a logistic regression model. RESULTS: Selective MMR nonreceipt, occurring in as few as 0.77% of children in the 1995 cohort, rose to 2.1% in the 2000 National Immunization Survey. Children included in the 2000 National Immunization Survey were born when the putative link between MMR and autism surfaced in the medical literature but before any significant media attention occurred. Selective nonreceipt was more prevalent in private practices and unrelated to family characteristics. MMR nonreceipt returned to baseline before sustained media coverage of the MMR-autism story began. CONCLUSIONS: There was a significant increase in selective MMR nonreceipt that was temporally associated with the publication of the original scientific literature, suggesting a link between MMR and autism, which preceded media coverage of the MMR-autism controversy. This finding suggests a limited influence of mainstream media on MMR immunization in the United States. PMID- 18381513 TI - Elevated morphine concentrations in neonates treated with morphine and prolonged hypothermia for hypoxic ischemic encephalopathy. AB - OBJECTIVES: Asphyxia and hypothermia may modify drug pharmacokinetics. We investigated whether analgesia with morphine in neonates with hypoxic ischemic encephalopathy undergoing prolonged moderate systemic hypothermia resulted in elevated serum morphine concentrations compared with normothermic infants. PATIENTS AND METHODS: Infants from 1 center participating in a multicenter randomized study of moderate whole-body hypothermia after perinatal asphyxia (the Total Body Hypothermia Study) were randomly selected for treatment with hypothermia (n = 10) or for standard care on normothermia (n = 6). Hypothermia (33 degrees C to 34 degrees C) was started before 6 hours of age and maintained for 72 hours. All of the infants were treated with a continuous infusion of morphine-hydrochloride, with the rate adjusted according to clinical status. Serum morphine concentrations were determined at 6, 12, 24, 48, and 72 hours after birth. RESULTS: Serum morphine concentrations at 24 to 72 hours after birth were (median [range]) 292 ng/mL (137-767 ng/mL) in the hypothermia-treated infants and 206 ng/mL (88-327 ng/mL) in the infants on normothermia, despite similar morphine infusion rates and cumulative doses. Morphine concentrations correlated with morphine infusion rate, cumulative dose, and treatment with hypothermia. Serum morphine concentrations reached a steady state after 24 hours in the normothermic infants but continued to increase throughout the assessment period in the hypothermia group. Morphine clearance was low in both groups: (median [range]) morphine clearance estimated from area under the curve was 0.69 mL/min per kg (0.58-1.21 mL/min per kg) in hypothermic group and 0.89 mL/min per kg (0.65-1.33 mL/min per kg) in infants on normothermia. Serum morphine concentrations >300 nL/mL occurred more often in the hypothermia group and when the morphine infusion rate was >10 microg/kg per h. CONCLUSIONS: Infants with hypoxic ischemic encephalopathy have reduced morphine clearance and elevated serum morphine concentrations when morphine infusion rates are based on clinical state. Potentially toxic serum concentrations of morphine may occur with moderate hypothermia and infusion rates >10 microg/kg per h. PMID- 18381514 TI - Prevention of vitamin K deficiency bleeding in breastfed infants: lessons from the Dutch and Danish biliary atresia registries. AB - OBJECTIVE: Newborns routinely receive vitamin K to prevent vitamin K deficiency bleeding. The efficacy of oral vitamin K administration may be compromised in infants with unrecognized cholestasis. We aimed to compare the risk of vitamin K deficiency bleeding under different prophylactic regimens in infants with biliary atresia. PATIENTS AND METHODS: From Dutch and Danish national biliary atresia registries, we retrieved infants who were either breastfed and received 1 mg of oral vitamin K at birth followed by 25 microg of daily oral vitamin K prophylaxis (Netherlands, 1991-2003), 2 mg of oral vitamin K at birth followed by 1 mg of weekly oral prophylaxis (Denmark, 1994 to May 2000), or 2 mg of intramuscular prophylaxis at birth (Denmark, June 2000-2005) or were fed by formula. We determined the absolute and relative risk of severe vitamin K deficiency and vitamin K deficiency bleeding on diagnosis in breastfed infants on each prophylactic regimen and in formula-fed infants. RESULTS: Vitamin K deficiency bleeding was noted in 25 of 30 of breastfed infants on 25 microg of daily oral prophylaxis, in 1 of 13 on 1 mg of weekly oral prophylaxis, in 1 of 10 receiving 2 mg of intramuscular prophylaxis at birth, and in 1 of 98 formula-fed infants (P < .001). The relative risk of a bleeding in breastfed compared with formula-fed infants was 77.5 for 25 microg of daily oral prophylaxis, 7.2 for 1 mg of weekly oral prophylaxis, and 9.3 for 2 mg of intramuscular prophylaxis at birth. CONCLUSIONS: A daily dose of 25 microg of vitamin K fails to prevent bleedings in apparently healthy infants with unrecognized cholestasis because of biliary atresia. One milligram of weekly oral prophylaxis offers significantly higher protection to these infants and is of similar efficacy as 2 mg of intramuscular prophylaxis at birth. Our data underline the fact that event analysis in specific populations at risk can help to evaluate and improve nationwide prophylactic regimens. PMID- 18381515 TI - Readmission for neonatal jaundice in California, 1991-2000: trends and implications. AB - OBJECTIVE: We sought to describe population-based trends, potential risk factors, and hospital costs of readmission for jaundice for term and late preterm infants. METHODS: Birth-cohort data were obtained from the California Office of Statewide Health Planning and Development and contained infant vital statistics data linked to infant and maternal hospital discharge summaries. The study population was limited to healthy, routinely discharged infants through the use of multiple exclusion criteria. All linked readmissions occurred within 14 days of birth. International Classification of Diseases, Ninth Revision, codes were used to further limit the sample to readmission for jaundice. Hospital discharge records were the source of diagnoses, hospital charges, and length-of-stay information. Hospital costs were estimated using hospital-specific ratios of costs to charges and adjusted to 1991. RESULTS: Readmission rates for jaundice generally rose after 1994 and peaked in 1998 at 11.34 per 1000. The readmission rate for late preterm infants (as a share of all infants) over the study period remained at <2 per 1000. Factors associated with increased likelihood of hospital readmission for jaundice included gestational age 34 to 39 weeks, birth weight of <2500 g, male gender, Medicaid or private insurance, and Asian race. Factors associated with a decreased likelihood of readmission for jaundice were cesarean section delivery and black race. The mean cost of readmission for all infants was $2764, with a median cost of $1594. CONCLUSIONS: Risk-adjusted readmission rates for jaundice rose following the 1994 hyperbilirubinemia guidelines and declined after postpartum length-of-stay legislation in 1998. In 2000, the readmission rate remained 6% higher than in 1991. These findings highlight the complex relationship among newborn physiology, socioeconomics, race or ethnicity, public policy, clinical guidelines, and physician practice. These trend data provide the necessary baseline to study whether revised guidelines will change practice patterns or improve outcomes. Cost data also provide a break-even point for prevention strategies. PMID- 18381516 TI - Neonatal dexamethasone treatment for chronic lung disease of prematurity alters the hypothalamus-pituitary-adrenal axis and immune system activity at school age. AB - OBJECTIVE: To compare long-term effects of neonatal treatment with dexamethasone or hydrocortisone for chronic lung disease of prematurity on the hypothalamus pituitary-adrenal axis and the immune response in children at school age. PATIENTS AND METHODS: A total of 156 prematurely born children were included in this retrospective matched cohort study. Children treated with dexamethasone (n = 52) or hydrocortisone (n = 52) were matched for gestational age, birth weight, grade of infant respiratory distress syndrome, grade of periventricular or intraventricular hemorrhage, gender, and year of birth. A reference group of 52 children not treated with corticosteroids was included for comparison. Plasma adrenocorticotropic hormone and cortisol in response to a social stress task were determined. Cytokine production was analyzed after in vitro stimulation of whole blood cultures. RESULTS: The Trier Social Stress Test adapted for children induced an adrenocorticotropic hormone and cortisol response in all of the groups. The adrenocorticotropic hormone response was blunted in the dexamethasone group. The overall cortisol level was lower in the dexamethasone than in the hydrocortisone and reference group. Cortisol and adrenocorticotropic hormone in the hydrocortisone and reference groups were similar. The ratio of T-cell mitogen induced interferon-gamma/interleukin-4 secretion was significantly higher in the dexamethasone group than in the hydrocortisone group. Interferon-gamma production and the ratios of interferon-gamma/interleukin-4 and interferon-gamma/ interleukin-10 were significantly higher in the dexamethasone group than the reference group. However, production of these cytokines did not differ between the hydrocortisone and the reference groups. CONCLUSION: Neonatal treatment of prematurely born children with dexamethasone but not with hydrocortisone resulted in long-lasting programming effects on hypothalamus-pituitary-adrenal axis and on the T-helper 1/T-helper 2 cytokine balance. Follow-up of these children is required to investigate long-term clinical consequences. We recommend that authors of previously performed randomized, controlled trials on neonatal glucocorticoid treatment include immune and neuroendocrine analyses in prolonged follow-up of these children. PMID- 18381517 TI - Genetics of pubertal timing and its associations with relative weight in childhood and adult height: the Swedish Young Male Twins Study. AB - OBJECTIVE: Previous studies have suggested that the timing of puberty is associated with BMI in childhood and adult stature. Because the genetic background of these associations is not thoroughly investigated, we aimed to analyze it in a longitudinal twin cohort. METHODS: We studied a Swedish cohort of 99 monozygotic and 76 dizygotic twin pairs born between 1973 and 1979 with weight and length or height measured annually from birth to age 18 years. Age at onset of pubertal growth spurt, age at peak height velocity, and final height were estimated by a parametric JPA2 growth model. The genetic architecture and mutual associations of these traits and childhood BMI were analyzed by linear structural equation modeling. RESULTS: The heritability estimate was 0.91 for age at onset of pubertal growth spurt, 0.93 for age at peak height velocity, and 0.97 for adult height. Age at onset of pubertal growth spurt was negatively associated with BMI from 1 to 10 years of age and stature in early adulthood. For age at peak height velocity, we found similar associations with childhood BMI and stature in early adulthood. These associations were explained by common genetic factors. CONCLUSION: Growth during puberty is strictly genetically regulated. These genetic factors also explain why boys who matured early had higher BMI through childhood and taller stature in early adulthood. PMID- 18381518 TI - Selective provision of asthma self-management tools to families. AB - OBJECTIVE: Providing asthma education in a primary care setting can be challenging because of time and resource constraints. The purpose of this work was to determine factors associated with the provision of different asthma self management tools. METHODS: We conducted a cross-sectional survey with 896 parents of children with asthma (age 2-12 years). We collected information regarding demographics and asthma care, including parent receipt of an asthma action plan, a symptom diary, and asthma information materials; whether an asthma management plan was sent to the child's school; and whether the physician reviewed written instructions on use of a metered-dose inhaler. We used multivariate logistic regression methods to determine factors associated with receipt of different asthma self-management tools controlling for demographic factors. RESULTS: For families where parents only completed high school, there was greater likelihood of receipt of an asthma action plan and physician review of written instructions about how to use an inhaler. For families with a household income less than twice the poverty line, there was greater likelihood of receipt of an asthma action plan, the physician sending a letter to the child's school regarding the child's asthma, and receipt of an asthma symptom diary. CONCLUSIONS: In our sample, primary care pediatricians do not routinely provide asthma education in accordance with National Heart, Lung, and Blood Institute asthma guidelines and "triage" which families receive additional asthma education. We believe that the use of targeted asthma education is a symptom of the limited time and competing demands during a typical visit. As a result, those involved in quality improvement need to help physicians become more efficient and effective at providing asthma education within such time constraints or develop alternative systems of providing asthma education. PMID- 18381519 TI - Comparison of critically ill and injured children transferred from referring hospitals versus in-house admissions. AB - OBJECTIVE: The purpose of this work was to compare the outcomes, severity of illness, and resource use of patients transferred to PICUs from outside hospitals to patients admitted from within the same hospital. METHODS: We conducted a secondary analysis of patients from the 20 US PICUs in the most recent Pediatric Intensive Care Unit Evaluations Software Recalibration Database on a total of 13,017 emergent PICU admissions between January 2001 and January 2006. Dependent variables were PICU resource use and risk-adjusted mortality. The main independent variable was the PICU admission source: patients transferred from referring emergency departments and inpatient wards versus in-house admissions from the same hospitals' emergency departments and inpatient ward. RESULTS: Patients admitted from referring emergency departments had higher use of vasoactive infusions (7.31% vs 5.23%) and mechanical ventilation (33.45% vs 23.6%) than same-hospital emergency department admissions. Compared with in-house ward admissions, patients transferred from referring inpatient wards had higher mechanical ventilation rates (45.05% vs 28.56%) and PICU lengths of stay (8.0 vs 6.7 days). CONCLUSIONS: On average, children admitted to a cohort of US PICUs from referring hospitals were more ill and required more intensive care resources than patients admitted to the same PICUs from within the institution. Hospital level differences in PICU efficiency and severity of illness were highly variable. These data highlight the need for standardized PICU admission criteria to maximize hospital efficiency and suggest opportunities for earlier intervention and consultation by hospitals with PICU-level services to improve quality of care for critically ill children. PMID- 18381520 TI - Self-management behaviors, racial disparities, and glycemic control among adolescents with type 2 diabetes. AB - OBJECTIVE: Type 2 diabetes is a growing problem among adolescents, but little is known about self-management behaviors in this population. Our aim was to examine self-management behaviors and glycemic control among adolescents with type 2 diabetes. METHODS: From 2003 to 2005, a telephone survey of adolescents with type 2 diabetes was performed. Chart review obtained most recent glycated hemoglobin and clinical characteristics. Analyses compared patient characteristics and self management behaviors to recent glycated hemoglobin levels. RESULTS: Of 139 patients contacted, 103 (74%) completed the study. The mean age was 15.4 years: 69% were girls, 47% were white, and 46% were black. Mean glycated hemoglobin was 7.7%, and the average duration of diabetes was 2.0 years. More than 80% of patients reported > or = 75% medication compliance, and 59% monitored blood glucose > 2 times daily. However, patients reported frequent episodes of overeating, drinking sugary drinks, and eating fast food. More than 70% of patients reported exercising > or = 2 times a week, but 68% reported watching > or = 2 hours of television daily. Nonwhite patients had higher glycated hemoglobin and hospitalizations per year compared with white patients. In multivariable analyses, nonwhite race remained significantly associated with higher glycated hemoglobin even after adjusting for age, gender, BMI, insurance status, and other factors. Nonwhite patients were more likely to watch > or = 2 hours of television per day (78% vs 56%), to report exercising < or = 1 time per week (35% vs 21%), and to drink > or = 1 sugary drink daily (27% vs 13%). CONCLUSION: Although patients reported good medication and monitoring adherence, they also reported poor diet and exercise habits and multiple barriers. Nonwhite race was significantly associated with poorer glycemic control even after adjusting for covariates. This may, in part, be related to disparities in lifestyle behaviors. Additional studies are indicated to further assess self management behaviors and potential racial disparities in adolescents with type 2 diabetes. PMID- 18381521 TI - Development, testing, and findings of a pediatric-focused trigger tool to identify medication-related harm in US children's hospitals. AB - OBJECTIVES: The purposes of this study were to develop a pediatric-focused tool for adverse drug event detection and describe the incidence and characteristics of adverse drug events in children's hospitals identified by this tool. METHODS: A pediatric-specific trigger tool for adverse drug event detection was developed and tested. Eighty patients from each site were randomly selected for retrospective chart review. All adverse drug events identified using the trigger tool were evaluated for severity, preventability, ability to mitigate, ability to identify the event earlier, and presence of associated occurrence report. Each trigger and the entire tool were evaluated for positive predictive value. RESULTS: Review of 960 randomly selected charts from 12 children's hospitals revealed 2388 triggers (2.49 per patient) and 107 unique adverse drug events. Mean adverse drug event rates were 11.1 per 100 patients, 15.7 per 1000 patient days, and 1.23 per 1000 medication doses. The positive predictive value of the trigger tool was 3.7%. Twenty-two percent of all adverse drug events were deemed preventable, 17.8% could have been identified earlier, and 16.8% could have been mitigated more effectively. Ninety-seven percent of the identified adverse drug events resulted in mild, temporary harm. Only 3.7% of adverse drug events were identified in existing hospital-based occurrence reports. The most common adverse drug events identified were pruritus and nausea, the most common medication classes causing adverse drug events were opioid analgesics and antibiotics, and the most common stages of the medication management process associated with preventable adverse drug events were monitoring and prescribing/ordering. CONCLUSIONS: Adverse drug event rates in hospitalized children are substantially higher than previously described. Most adverse drug events resulted in temporary harm, and 22% were classified as preventable. Only 3.7% were identified by using traditional voluntary reporting methods. Our pediatric-focused trigger tool is effective at identifying adverse drug events in inpatient pediatric populations. PMID- 18381522 TI - Variability in expert assessments of child physical abuse likelihood. AB - OBJECTIVES: In the absence of a gold standard, clinicians and researchers often categorize their opinions of the likelihood of inflicted injury using several ordinal scales. The objective of this protocol was to determine the reliability of expert ratings using several of these scales. METHODS: Participants were pediatricians with substantial academic and clinical activity in the evaluation of children with concerns for physical abuse. The facts from several cases that were referred to 1 hospital's child abuse team were abstracted and recorded as in a multidisciplinary team conference. Participants viewed the recording and rated each case using several scales of child abuse likelihood. RESULTS: Participants (n = 22) showed broad variability for most cases on all scales. Variability was lowest for cases with the highest aggregate concern for abuse. One scale that included examples of cases fitting each category and standard reporting language to summarize results showed a modest (18%-23%) decrease in variability among participants. The interpretation of the categories used by the scales was more consistent. Cases were rarely rated as "definite abuse" when likelihood was estimated at < or = 95%. Only 7 of 156 cases rated < or = 15% likelihood were rated as "no reasonable concern for abuse." Only 9 of 858 cases rated > or = 35% likelihood were rated as "reasonable concern for abuse." CONCLUSIONS: Assessments of child abuse likelihood often show broad variability between experts. Although a rating scale with patient examples and standard reporting language may decrease variability, clinicians and researchers should be cautious when interpreting abuse likelihood assessments from a single expert. These data support the peer review or multidisciplinary team approach to child abuse assessments. PMID- 18381523 TI - Gymnastics-related injuries to children treated in emergency departments in the United States, 1990-2005. AB - OBJECTIVE: The objective of this study was to describe the epidemiology of gymnastics-related injuries among children in the United States. METHODS: A retrospective analysis was conducted of data for children 6 through 17 years of age from the National Electronic Injury Surveillance System of the US Consumer Product Safety Commission for 1990 through 2005. Sample weights provided by the National Electronic Injury Surveillance System were used to calculate national estimates of gymnastics-related injuries. Injury rates were calculated for the most frequently occurring types of injury using gymnastics participation data. RESULTS: An estimated 425,900 children 6 through 17 years of age were treated in US hospital emergency departments for gymnastics-related injuries during the 16 year period of 1990-2005. The number of injuries averaged 26,600 annually. The number of injuries sustained per 1000 gymnastics participants per year for the pediatric population was 4.8. The number of injuries sustained per 1000 participants per year was greater for the ages 12 to 17 years (7.4) than for the ages 6 to 11 years (3.6). The place where an injury occurred include school (40.0%), a place of recreation or sports (39.7%), home (14.5%), or on other public property (5.8%). The body parts injured were upper extremity (42.3%), lower extremity (33.8%), head/neck (12.9%), trunk (10.4%), and other (0.6%). Injury diagnoses were strain/sprain (44.5%), fracture/dislocation (30.4%), abrasion/contusion (15.6%), laceration/avulsion (3.7%), concussion/closed head injury (1.7%), and other (4.2%). The majority (97.1%) of patients with gymnastics related injuries were treated and released from the emergency department. CONCLUSIONS: Gymnastics has one of the highest injury rates of all girls' sports. Establishment of a national database for gymnastics-related injuries, including exposure data for direct calculation of injury rates, would permit better identification and monitoring of risk factors for gymnastics-related injuries and aid in the development, implementation, and evaluation of injury prevention strategies based on epidemiologic evidence. PMID- 18381524 TI - Growth in stimulant-naive children with attention-deficit/hyperactivity disorder using cross-sectional and longitudinal approaches. AB - OBJECTIVE: The objective of this study was to investigate the growth of stimulant naive children with attention-deficit/hyperactivity disorder in 3 aspects of development: level, trend, and structure of the process. PATIENTS AND METHOD: The study sample included 53 individuals between the ages of 6 and 17 years (mean: 11.90). The average level of growth (z scores) in prepubertal, pubertal, and postpubertal stage of development and trend of the process were estimated on the basis of a comparison with growth charts. The prediction of adolescent growth spurt was conducted using the mathematical structural growth model of Jolioceur, Pontier, and Abidi. RESULTS: Difference between boys with attention deficit/hyperactivity and the norm was statistically significant in the prepubertal stage and for the average level of growth between the ages of 2 and 17 years. Distinct suppression of growth was found between the ages of 9 and 14. Analysis of development structure revealed an earlier onset of the adolescent growth spurt among boys (difference: 5 months) and a higher velocity of growth at this moment (difference: 0.33 cm/year) than expected values. CONCLUSIONS: The application of both cross-sectional and longitudinal analyses clearly illustrated the higher level of growth of boys with attention-deficit/hyperactivity in the prepubertal stage, the suppression of growth on the turn of prepubertal and pubertal periods, and earlier occurrence of the spurt onset. Observed differences in the level and trend of growth as well as in the parameters of adolescent growth spurt are linked with disorder-related factors. PMID- 18381525 TI - Use of aromatase inhibitors in children and adolescents with disorders of growth and adolescent development. AB - Although treatment of children and adolescents who have disorders of growth and adolescent development with aromatase inhibitors is increasingly common, data for or against their use are extremely limited. Precocious puberty, short stature, and gynecomastia are conditions for which inhibition of the enzyme aromatase might prove beneficial to reduce clinical signs of estrogenization and/or estrogen-mediated skeletal maturation. In this report, we summarize the published data regarding the use of aromatase inhibitors in these conditions, and review known and potential benefits, safety concerns, and shortcomings of the available information. PMID- 18381526 TI - Global initiatives for improving hospital care for children: state of the art and future prospects. AB - Deficiencies in the quality of health care are major limiting factors to the achievement of the Millennium Development Goals for child and maternal health. Quality of patient care in hospitals is firmly on the agendas of Western countries but has been slower to gain traction in developing countries, despite evidence that there is substantial scope for improvement, that hospitals have a major role in child survival, and that inequities in quality may be as important as inequities in access. There is now substantial global experience of strategies and interventions that improve the quality of care for children in hospitals with limited resources. The World Health Organization has developed a toolkit that contains adaptable instruments, including a framework for quality improvement, evidence-based clinical guidelines in the form of the Pocket Book of Hospital Care for Children, teaching material, assessment, and mortality audit tools. These tools have been field-tested by doctors, nurses, and other child health workers in many developing countries. This collective experience was brought together in a global World Health Organization meeting in Bali in 2007. This article describes how many countries are achieving improvements in quality of pediatric care, despite limited resources and other major obstacles, and how the evidence has progressed in recent years from documenting the nature and scope of the problems to describing the effectiveness of innovative interventions. The challenges remain to bring these and other strategies to scale and to support research into their use, impact, and sustainability in different environments. PMID- 18381527 TI - Basic training for the pediatrician: how to provide comprehensive anticipatory guidance regarding military service. PMID- 18381528 TI - Age inconsistency in the American Academy of Pediatrics guidelines for acute otitis media. AB - OBJECTIVE: The American Academy of Pediatrics acute otitis media guidelines could reduce antibiotic use. The objective was to compare strategies for diagnosing and treating otitis: (1) a commonly used, 2-criteria strategy, (2) the guidelines' 3 criteria algorithm, and (3) initially watching without antibiotics. METHODS: A decision analysis was performed with literature-based parameter. The target population was children presenting to primary care physicians with possible otitis media. Main outcomes were antibiotic use, sick days, mild adverse drug events, and number needed to treat/avoided sick day. RESULTS: For children 2 to <6 months of age, compared with the 2-criteria strategy, guideline use predicted 21% less antibiotic use, 13% more sick days, and 23% fewer adverse drug events; the number needed to treat for the 2-criteria strategy versus the American Academy of Pediatrics strategy was 1.2 children per avoided sick day. For children 6 to <24 months of age, guideline use, compared with the 2-criteria strategy, predicted 26% less antibiotic use, 14% more sick days, and 28% fewer adverse drug events; the number needed to treat for the 2-criteria strategy versus the American Academy of Pediatrics strategy was 1.4 children per avoided sick day. For children >2 years of age, guideline use, compared with the 2 criteria strategy, predicted 67% less antibiotic use, 4% more sick days, and 68% fewer adverse drug events. The number needed to treat for the guideline strategy versus the watch strategy was 6.3 children per avoided sick day; that for the 2 criteria strategy versus the guideline strategy was 12.3. Guideline use for children <2 years implies that our number needed to treat to avoid a sick day is <1.4; for children >2, guideline use implies we are willing to treat at least 6.3 children to avoid a sick day. Thus, the guidelines imply a greater willingness to treat older children, compared with younger children. CONCLUSIONS: The American Academy of Pediatrics guidelines are inconsistent in their outcomes across age groups. Guideline implementation under age 2 reduces antibiotic use but at a relatively heavy cost of sick days and parental missed work days. This trade-off may be particularly unfavorable for working parents, who might reasonably prefer greater antibiotic use. PMID- 18381529 TI - Valuing reduced antibiotic use for pediatric acute otitis media. AB - OBJECTIVE: The 2004 American Academy of Pediatrics acute otitis media guidelines urge parents to weigh the benefits of reduced antibiotic use, adverse drug events, and future resistance versus risks of extra costs and sick days resulting from guideline use. The value of decreased antibiotic resistance has not been quantified. The objective was to perform cost-utility analysis, estimating the resistance value of implementing the guidelines for acute otitis media treatment for children <2 years of age. Outcomes were described with a common denominator and the value of avoiding resistance was estimated using a parental perspective. METHODS: Decision analysis results were used for outcome probabilities. Published utilities were used to describe outcomes in quality-adjusted life-day units. The minimum resistance benefit value, where the benefits of the American Academy of Pediatrics guidelines would at least balance their costs, was defined as the guidelines' incremental costs minus their other benefits. RESULTS: For a child 2 to <6 months of age presenting to a primary care physician with possible otitis media, parents would need to value the resistance benefit at 0.77 quality adjusted life-days per antibiotic prescription avoided for the guidelines' benefits to balance their costs. For the 6- to <24-month-old group, results were 0.67 quality-adjusted life-days per prescription avoided. Results were sensitive to the dollar cost utility; when willingness to pay ranged from $20,000 to $200,000 per quality-adjusted life-year, results ranged from 0.36 and 0.30 quality-adjusted life-days up to 4.10 and 3.57 quality-adjusted life-days for the 2- to <6-month-old and 6- to <24-month-old groups, respectively. Costs were driven by missed parent work days. CONCLUSIONS: From a societal perspective, trading 0.30 to 4 quality-adjusted life-days to avoid 1 antibiotic course might be desirable; from a parental perspective, this may not be as desirable. Parent demand for antibiotics may be rational when driven by the value of parent time. Other approaches that have the potential to reduce antibiotic use, such as wider use of influenza vaccine and improved rapid viral diagnostic techniques, might be more successful. PMID- 18381530 TI - Trends in otitis media treatment failure and relapse. AB - OBJECTIVES: The goals were to describe trends in acute otitis media, treatment failure, and relapse and in high-dose amoxicillin use and to determine whether treatment of acute otitis media with high-dose amoxicillin was associated with treatment failure or relapse. METHODS: We conducted a retrospective study of acute otitis media visits made between 1996 and 2004 by children 2 months to 12 years of age in a large group practice, using computerized data. We defined acute otitis media as an otitis media visit with antibiotics dispensed (preceded by 30 days without otitis media visits), treatment failure as initiation of treatment with a second antibiotic before the first prescription was finished, and relapse as initiation of antibiotic treatment after the first prescription was finished but within 30 days after the index acute otitis media episode. The primary independent measure was high-dose amoxicillin (>70 mg/kg per day). We evaluated changes over time and determined whether high-dose amoxicillin use was associated with otitis media treatment failure or relapse. RESULTS: We identified 111,335 acute otitis media visits over a 9-year period. The incidence of acute otitis media decreased from 385.1 visits per 1000 enrollees in 1996 to 188.8 visits per 1000 enrollees in 2004. The proportion of acute otitis media visits treated with high-dose amoxicillin increased from 1.7% in 1996 to 41.9% in 2004. Both otitis media treatment failure and relapse rates decreased from 1996 to 2004 (from 3.9% to 2.6% and from 9.2% to 8.9%, respectively). The odds of treatment failure or relapse did not differ between acute otitis media episodes treated with high-dose and low-dose amoxicillin. CONCLUSIONS: During the past decade, acute otitis media, treatment failure, and relapse became less common and high-dose amoxicillin use increased. However, high-dose amoxicillin treatment did not reduce the risk of individual infections resulting in adverse outcomes. PMID- 18381531 TI - Prospective multicenter study of bronchiolitis: predicting safe discharges from the emergency department. AB - OBJECTIVE: Bronchiolitis is the leading cause of hospitalization for infants. Our objective was to identify factors associated with safe discharge to home from the emergency department. METHODS: We conducted a prospective cohort study during 2 consecutive bronchiolitis seasons, from 2004 to 2006. Thirty US emergency departments contributed data. All patients were < 2 years of age and had a final emergency department attending physician diagnosis of bronchiolitis. Using multivariate logistic regression, a low-risk model was developed with a random half of the data and then validated with the other half. RESULTS: Of 1456 enrolled patients, 837 (57%) were discharged home from the emergency department. The following factors predicted safe discharge to home: age of > or = 2 months, no history of intubation, a history of eczema, age-specific respiratory rates (< 45 breaths per minute for 0-1.9 months, < 43 breaths per minute for 2-5.9 months, and < 40 breaths per minute for 6-23.9 months), no/mild retractions, initial oxygen saturation of > or = 94%, fewer albuterol or epinephrine treatments in the first hour, and adequate oral intake. The importance of each factor varied slightly according to age, but the comprehensive model (developed and validated for all children < 2 years of age) yielded an area under the receiver operating characteristic curve of 0.81, with a good fit of the data. CONCLUSIONS: This large multicenter study of children presenting to the emergency department with bronchiolitis identified several factors associated with safe discharge, including cut points for respiratory rate and oxygen saturation. Although the low risk model requires further study, we believe that it will assist clinicians evaluating children with bronchiolitis and may help reduce some unnecessary hospitalizations. PMID- 18381532 TI - Changing patterns in neonatal Escherichia coli sepsis and ampicillin resistance in the era of intrapartum antibiotic prophylaxis. AB - OBJECTIVE: The goal was to determine current trends in Escherichia coli-related early- and late-onset sepsis and patterns of ampicillin resistance in relation to institutional changes in the use of intrapartum antibiotic prophylaxis. METHODS: A retrospective review of data for all infants with E. coli sepsis at Yale-New Haven Hospital from 1979 to 2006 was performed. Study periods were based on predominant intrapartum antibiotic prophylaxis practices at Yale-New Haven Hospital, that is, (1) 1979 to 1992 (no formal intrapartum antibiotic prophylaxis), (2) 1993 to 1996 (risk factor-based), and (3) 1997 to 2006 (screening-based). Sepsis rates and patterns of ampicillin resistance were compared. RESULTS: Fifty-three cases of E. coli early-onset sepsis and 129 cases of E. coli late-onset sepsis were identified over 3 eras. In very low birth weight (<1500 g) infants, increases in E. coli early-onset sepsis (period 1: 2.83 cases per 1000 very low birth weight admissions; period 2: 7.12 cases per 1000 very low birth weight admissions; period 3: 10.22 cases per 1000 very low birth weight admissions), intrapartum ampicillin exposure, and ampicillin-resistant E. coli were observed. Intrapartum ampicillin exposure was determined to be an independent risk factor for ampicillin-resistant E. coli early-onset sepsis. For the first time, a significant increase in E. coli late-onset sepsis was observed in preterm infants (period 1: 10.39 cases per 1000 very low birth weight admissions; period 2: 16.01 cases per 1000 very low birth weight admissions; period 3: 21.66 cases per 1000 very low birth weight admissions) and term infants (period 1: 4.07 cases per 1000 admissions; period 2: 4.22 cases per 1000 admissions; period 3: 8.23 cases per 1000 admissions). CONCLUSIONS: Studies to provide a better understanding of potential consequences of intrapartum antibiotic exposure and its contribution to evolving trends in neonatal sepsis are urgently needed. PMID- 18381533 TI - Neonatal antibiotic treatment is a risk factor for early wheezing. AB - OBJECTIVE: The use of antibiotics in infancy and subsequent changes in the intestinal bacterial flora have been discussed as risk factors for the development of asthma. However, it has been difficult to exclude the possibility that antibiotics have been given in early episodes of wheezing. As a result, there has been a risk of reverse causation. To minimize the risk of reverse causation, we have focused on the effect of antibiotics that are already administered on the neonatal ward. METHODS: In a cohort study of infants born in western Sweden in 2003, we studied the development of wheezing. The families of the infants were randomly selected and sent a questionnaire at child ages 6 and 12 months. The response rate was 68.5% to the 6-month questionnaire and 68.9% to the 12-month questionnaire. RESULTS: At 12 months, 20.2% of infants had had 1 or more episodes of wheezing, and 5.3% had had 3 or more episodes. Inhaled corticosteroids had been taken by 4.1% of the infants. Independent risk factors for wheezing disorder treated with inhaled corticosteroids were neonatal antibiotic treatment, male gender, gestational age of <37 weeks, having a mother with asthma, having a sibling with asthma or eczema, and breastfeeding for <5 months. CONCLUSIONS: Treatment with antibiotics in the neonatal period was an independent risk factor for wheezing that was treated with inhaled corticosteroids at 12 months of age. These results indirectly support the hypothesis that an alteration in the intestinal flora can increase the risk of subsequent wheezing. PMID- 18381534 TI - Fluconazole prophylaxis in extremely low birth weight neonates reduces invasive candidiasis mortality rates without emergence of fluconazole-resistant Candida species. AB - OBJECTIVE: We evaluated the impact of fluconazole prophylaxis for extremely low birth weight infants on invasive candidiasis incidence, invasive candidiasis related mortality rates, and fluconazole susceptibility of Candida isolates. METHODS: Extremely low birth weight infants <5 days of age, except those with liver dysfunction, were eligible for fluconazole prophylaxis. NICU infants (all birth weights) with invasive candidiasis between April 2002 and March 2006 were compared with those with invasive candidiasis before fluconazole prophylaxis (2000-2001). RESULTS: Twenty-two infants had invasive candidiasis (all candidemia) during fluconazole prophylaxis; before fluconazole prophylaxis, there were 19 cases (candidemia: 17 cases; meningitis: 2 cases). Invasive candidiasis incidence in NICU infants decreased from 0.6% (19 of 3012 infants) before fluconazole prophylaxis to 0.3% (22 of 6393 infants) in 2002-2006 and that in extremely low birth weight infants decreased 3.6-fold. No Candida-attributable deaths occurred during 2002-2006 fluconazole prophylaxis, compared with 4 (21%) before fluconazole prophylaxis. The onset of invasive candidiasis was later during 2002-2006 (23.5 vs 12 days), but risk factors were similar. The invasive candidiasis species distribution remained stable. Of 409 infants who received fluconazole prophylaxis, 119 (29%) received 42 days. Shorter fluconazole prophylaxis duration was related to intravenous access no longer being necessary in 242 cases (59%), noninvasive candidiasis-related death in 29 (7%), hospital transfer in 8 (2%), invasive candidiasis diagnosis in 8 (2%), and transient increase in serum transaminase levels in 4 (1%). One hundred twenty-seven infants (31%) who received fluconazole prophylaxis developed cholestasis during hospitalization, two thirds of whom had other predisposing conditions. On multivariate logistic regression necrotizing enterocolitis and increasing days of total parenteral nutrition, but not increasing number of doses on days of fluconazole, were significantly associated with the development of cholestasis. CONCLUSION: During 4 years of fluconazole prophylaxis, the incidence of invasive candidiasis and invasive candidiasis-associated mortality rates in extremely low birth weight infants were reduced significantly, without the emergence of fluconazole-resistant Candida species. PMID- 18381535 TI - Characteristics of general and subspecialty pediatricians who choose not to recertify. AB - OBJECTIVE: The goal was to determine the reasons given for nonparticipation in maintenance of certification by general pediatricians and pediatric subspecialists whose board certification had expired. METHODS: All United States based pediatricians whose board certification had expired and who had not recertified as a generalist or subspecialist in 2004 or 2005 were surveyed. The total sample included 1001 pediatric generalists and 1237 subspecialists. We conducted a mail survey study, with a total of 3 mailings to nonrespondents. Analyses initially were conducted separately for generalists with expired certificates, subspecialists with active subspecialty but expired general pediatrics certificates, and subspecialists with expired subspecialty certificates (regardless of their general pediatrics certification status). RESULTS: The overall response rates were 68% for generalists and 76% for subspecialists. Of pediatricians who had allowed their certificates to expire, the majority still intended to recertify (65% of the generalists and 86% of the subspecialists, in their primary subspecialty). The most common reasons cited by the 35% of generalists with expired certificates who were not planning to participate in maintenance of certification were the expense, the time required to complete maintenance of certification, and the perceived lack of relevance to their current practice. The 14% of subspecialists who were not planning to recertify in their primary subspecialty most commonly cited the expense, a change in career path making recertification unnecessary, and the time required. CONCLUSIONS: Although the physicians in this study had allowed their certificates to expire, they still overwhelmingly believed that physicians who provide direct patient care should maintain their certification. There was general consensus among our respondents regarding the professional value to patients and peers of participation in the maintenance of certification program. PMID- 18381536 TI - Characteristics associated with older adolescents who have a television in their bedrooms. AB - OBJECTIVES: The goals were to examine the prevalence of adolescents having a television in their bedroom and to describe associated personal, social, and behavioral characteristics. METHODS: Participants included 781 adolescents (mean age: 17.2 years) who completed a mailed Project Eating Among Teens II questionnaire. The relationships between adolescents having a television in their bedroom and sociodemographic, behavioral, and personal characteristics were examined. RESULTS: Nearly two thirds (62%) of participants had a bedroom television. Gender, race/ethnicity, socioeconomic status, and age were associated with the presence of a bedroom television. Compared with girls without a bedroom television, girls with a bedroom television reported less time spent in vigorous activity (1.8 vs 2.5 hours/week), more time spent watching television (20.7 vs 15.2 hours/week), lower vegetable intake (1.7 vs 2.0 servings per day), greater sweetened beverage consumption (1.2 vs 1.0 servings per day), and fewer family meals (2.9 vs 3.7 meals per week). Compared with boys without a bedroom television, boys with a bedroom television reported more time spent watching television (22.2 vs 18.2 hours/week), lower fruit intake (1.7 vs 2.2 servings per day), fewer family meals (2.9 vs 3.6 meals per week), and lower grade point average (2.6 vs 2.9). Twice as many youths with a television in their bedroom were heavy television users (watched >5 hours/day), compared with youths without a television in their bedroom (16% vs 8%). CONCLUSIONS: Adolescents with a bedroom television reported more television viewing time, less physical activity, poorer dietary habits, fewer family meals, and poorer school performance. Refraining from placing a television in teenagers' rooms may be a first step in helping to decrease screen time and subsequent poor behaviors associated with increased television watching. PMID- 18381537 TI - Importance of early neglect for childhood aggression. AB - OBJECTIVE: The goal was to examine the association between early childhood neglect (birth to age 2 years) and later childhood aggression at ages 4, 6, and 8 years, compared with aggression's associations with early childhood abuse and later abuse and neglect. METHODS: A prospective cohort of 1318 predominantly at risk children, recruited from 4 US cities and 1 southern state, were monitored from birth to 8 years of age. Maltreatment was determined through review of local child protective services records. A hierarchical, linear model approach, a special case of general, linear, mixed modeling, was used to predict aggressive behavior scores, as reported by the child's primary caregiver at ages 4, 6, and 8 years. RESULTS: Only early neglect significantly predicted aggression scores. Early abuse, later abuse, and later neglect were not significantly predictive in a controlled model with all 4 predictors. CONCLUSION: This longitudinal study suggests that child neglect in the first 2 years of life may be a more-important precursor of childhood aggression than later neglect or physical abuse at any age. PMID- 18381538 TI - Just, in time: ethical implications of serial predictions of death and morbidity for ventilated premature infants. AB - OBJECTIVES: For a cohort of extremely premature, ventilated, newborn infants, we determined the power of either serial caretaker intuitions of "die before discharge" or serial illness severity scores to predict the outcomes of death in the NICU or neurologic performance at corrected age of 2 years. METHODS: We identified 268 premature infants who were admitted to our NICU in 1999-2004 and required mechanical ventilation. For each infant on each day of mechanical ventilation, we asked nurses, residents, fellows, and attending physicians the following question: "Do you think this child is going to live to go home or die before hospital discharge?" In addition, we calculated illness severity scores until either death or extubation. RESULTS: A total of 17,066 intuition profiles were obtained on 5609 days of mechanical ventilation in the NICU. One hundred (37%) of 268 profiled infants had > or = 1 intuition of die before discharge. Only 33 infants (33%) with an intuition of die actually died in the NICU. Of 48 infants with even 1 day of corroborated intuition of die in the NICU, only 7 (14%) were alive with both Mental Developmental Index and Psychomotor Developmental Index scores of > 69, and only 2 (4%) were alive with both Mental Developmental Index and Psychomotor Developmental Index Scores of > 79 at corrected age of 2 years. On day of life 1, the Score for Neonatal Acute Physiology II value for nonsurvivors (38.2 +/- 18.1) was significantly higher than that for survivors (26.3 +/- 12.7). However, this difference decreased steadily over time as scores improved for both groups. CONCLUSIONS: Illness severity scores become progressively less helpful over time in distinguishing infants who will either die in the NICU or survive with low Mental Developmental Index/Psychomotor Developmental Index scores. Serial caretaker intuitions of die before discharge also fail to identify prospective nonsurviving infants. However, corroborated intuitions of die before discharge identify a subset of infants whose likelihood of surviving to 2 years with both MDI and PDI > 80 is approximately 4%. PMID- 18381539 TI - Volumetric MRI study of brain in children with intrauterine exposure to cocaine, alcohol, tobacco, and marijuana. AB - OBJECTIVE: The objective of this study was to use volumetric MRI to study brain volumes in 10- to 14-year-old children with and without intrauterine exposure to cocaine, alcohol, cigarettes, or marijuana. METHODS: Volumetric MRI was performed on 35 children (mean age: 12.3 years; 14 with intrauterine exposure to cocaine, 21 with no intrauterine exposure to cocaine) to determine the effect of prenatal drug exposure on volumes of cortical gray matter; white matter; subcortical gray matter; cerebrospinal fluid; and total parenchymal volume. Head circumference was also obtained. Analyses of each individual substance were adjusted for demographic characteristics and the remaining 3 prenatal substance exposures. RESULTS: Regression analyses adjusted for demographic characteristics showed that children with intrauterine exposure to cocaine had lower mean cortical gray matter and total parenchymal volumes and smaller mean head circumference than comparison children. After adjustment for other prenatal exposures, these volumes remained smaller but lost statistical significance. Similar analyses conducted for prenatal ethanol exposure adjusted for demographics showed significant reduction in mean cortical gray matter; total parenchymal volumes; and head circumference, which remained smaller but lost statistical significance after adjustment for the remaining 3 exposures. Notably, prenatal cigarette exposure was associated with significant reductions in cortical gray matter and total parenchymal volumes and head circumference after adjustment for demographics that retained marginal significance after adjustment for the other 3 exposures. Finally, as the number of exposures to prenatal substances grew, cortical gray matter and total parenchymal volumes and head circumference declined significantly with smallest measures found among children exposed to all 4. CONCLUSIONS; These data suggest that intrauterine exposures to cocaine, alcohol, and cigarettes are individually related to reduced head circumference; cortical gray matter; and total parenchymal volumes as measured by MRI at school age. Adjustment for other substance exposures precludes determination of statistically significant individual substance effect on brain volume in this small sample; however, these substances may act cumulatively during gestation to exert lasting effects on brain size and volume. PMID- 18381540 TI - Epidemiologic features of the presentation of critical congenital heart disease: implications for screening. AB - OBJECTIVE: Critical congenital heart disease has been proposed as a target of newborn screening. This study aimed to define the incidence and timing of significant physiologic compromise attributable to critical congenital heart disease as well as the distribution of vulnerable lesions. These descriptive parameters must be defined to evaluate the impact and feasibility of any proposed screening strategy. METHODS: A retrospective cohort study of neonates who had critical congenital heart disease and were admitted to a single institution was conducted. Critical congenital heart disease was defined as congenital heart disease that required invasive intervention or resulted in death in the first 30 days of life. Significant physiologic compromise was defined by severe metabolic acidosis, seizure, cardiac arrest, or laboratory evidence of renal or hepatic injury before invasive intervention. Significant physiologic compromise was classified as potentially preventable when it occurred as a result of undiagnosed congenital heart disease after 12 hours of life. RESULTS: Significant physiologic compromise occurred in 76 (15.5%) of 490 patients, and potentially preventable significant physiologic compromise occurred in 33 (6.7%) of 490 patients. Most (83%) significant physiologic compromise as a result of unrecognized congenital heart disease occurred after 12 hours of age. A total of 90.9% of cases of potentially preventable significant physiologic compromise had aortic arch obstruction. The incidence of potentially preventable significant physiologic compromise as a result of congenital heart disease in the general population is estimated to be 1 per 15,000 to 1 per 26,000 live births. CONCLUSIONS: The incidence and timing of significant physiologic compromise as a result of critical congenital heart disease seems amenable to postnatal screening. Any viable screening strategy must be sensitive for lesions with aortic arch obstruction. PMID- 18381541 TI - Positive screening for autism in ex-preterm infants: prevalence and risk factors. AB - OBJECTIVE: The survival of very low birth weight infants has increased markedly in recent years. Unfortunately, the prevalence of significant and lifelong motor, cognitive, and behavioral dysfunction has remained a major problem confronting these children. The objective of this study was to perform screening tests for early autistic features in children with a history of very low birth weight and to identify risk factors associated with a positive screening result. METHODS: We studied 91 ex-preterm infants < or = 1500 g at birth. Infants underwent conventional MRI studies at preterm and/or term-adjusted age. We collected pertinent demographic, prenatal, intrapartum, acute postnatal, and short-term outcome data for all infants. Follow-up assessments were performed at a mean age of 21.9 +/- 4.7 months, using the Modified Checklist for Autism in Toddlers, the Vineland Adaptive Behavior Scale, and the Child Behavior Checklist. RESULTS: Twenty-six percent of ex-preterm infants had a positive result on the autism screening tool. Abnormal scores correlated highly with internalizing behavioral problems on the Child Behavior Checklist and socialization and communication deficits on the Vineland Scales. Lower birth weight, gestational age, male gender, chorioamnionitis, acute intrapartum hemorrhage, illness severity on admission, and abnormal MRI studies were significantly associated with an abnormal autism screening score. CONCLUSIONS: Early autistic behaviors seem to be an underrecognized feature of very low birth weight infants. The results from this study suggest that early screening for signs of autism may be warranted in this high-risk population followed by definitive autism testing in those with positive screening results. PMID- 18381542 TI - Growth and nutrient intakes of human milk-fed preterm infants provided with extra energy and nutrients after hospital discharge. AB - OBJECTIVES: The purpose of this pilot study was to determine whether mixing a multinutrient fortifier to approximately one half of the human milk fed each day for a finite period after discharge improves the nutrient intake and growth of predominantly human milk-fed low birth weight infants. We also assessed the impact of this intervention on the exclusivity of human milk feeding. METHODS: Human milk-fed (> or = 80% feeding per day) low birth weight (750-1800 g) infants (n = 39) were randomly assigned at hospital discharge to either a control or an intervention group. Infants in the control group were discharged from the hospital on unfortified human milk. Nutrient enrichment of human milk in the intervention group was achieved by mixing approximately one half of the human milk provided each day with a powdered multinutrient human milk fortifier for 12 weeks after discharge. Milk with added nutrients was estimated to contain approximately 80 kcal (336 kJ) and 2.2 g protein/100 mL plus other nutrients. Intensive lactation support was provided to both groups. RESULTS: Infants in the intervention group were longer during the study period, and those born < or = 1250 g had larger head circumferences than infants in the control group. There was a trend toward infants in the intervention group to be heavier at the end of the intervention compared with those in the control group. Mean protein, zinc, calcium, phosphorus, and vitamins A and D intakes were higher in the intervention group. CONCLUSIONS: Results from this study suggest that adding a multinutrient fortifier to approximately one half of the milk provided to predominantly human milk-fed infants for 12 weeks after hospital discharge may be an effective strategy in addressing early discharge nutrient deficits and poor growth without unduly influencing human milk feeding when intensive lactation support is provided. PMID- 18381543 TI - Prevalence and risk factors of elevated pulmonary artery pressures in children with sickle cell disease. AB - OBJECTIVES: The objectives of this study were (1) to determine the prevalence and risk factors of elevated pulmonary artery pressures in children with homozygous SS or Sbeta(0) thalassemia using Doppler echocardiography and (2) to determine a correlation between abnormal transcranial Doppler examinations and elevated pulmonary artery pressures. METHODS: Screening echocardiograms were prospectively performed during an annual comprehensive clinic visit on children who were older than 6 years and had homozygous SS or Sbeta(0) thalassemia. Detailed history, examination, and laboratory tests were done, and transcranial Doppler examinations were obtained in children 2 to 14 years of age. Pulmonary hypertension was defined as pulmonary artery systolic pressure of at least 30 mmHg corresponding to a peak tricuspid regurgitant jet velocity of > or = 2.5 m/second. Mild pulmonary hypertension was defined as tricuspid regurgitant jet velocity > or = 2.5 to 2.9 m/second. Moderate pulmonary hypertension was defined as tricuspid regurgitant jet velocity > or = 3 m/second. Patients with pulmonary stenosis or right outflow obstruction were excluded. Characteristics were compared between patients with mild, moderate, and no pulmonary hypertension using 1-way analysis of variance for continuous variable and Fisher's exact test for categorical variables. RESULTS: Of the 75 patients who had homozygous SS/Sbeta(0) thalassemia and were older than 6 years, echocardiograms were obtained for 62 (82.6%). Thirty percent (19 of 62) of patients had elevated tricuspid regurgitant jet velocity > or = 2.5 m/second. One third of these patients had tricuspid regurgitant jet velocity > or = 3 m/second. All patients with elevated tricuspid regurgitant jet velocity had SS disease. A high reticulocyte count, low oxygen saturation, and a high platelet count were significantly associated with elevated pulmonary artery pressures. There was no difference in age, gender, history of acute chest syndrome, hydroxyurea therapy, chronic blood transfusion, stroke, hemoglobin, and bilirubin between patients with and without elevated pulmonary artery pressures. A total of 47% patients with elevated tricuspid regurgitant jet velocity and 57% without elevated tricuspid regurgitant jet velocity had screening transcranial Doppler examinations. Transcranial Doppler examinations were normal for all patients. CONCLUSIONS: High pulmonary artery pressures do occur in children with sickle cell disease. Screening by echocardiography can lead to early detection and intervention that may potentially reverse this disease process. There was no correlation between elevated pulmonary artery pressures and abnormal transcranial Doppler examination in our study. PMID- 18381544 TI - Annual summary of vital statistics: 2006. AB - US births increased 3% between 2005 and 2006 to 4,265,996, the largest number since 1961. The crude birth rate rose 1%, to 14.2 per 1000 population, and the general fertility rate increased 3%, to 68.5 per 1000 women 15 to 44 years. Births and birth rates increased among all race and Hispanic-origin groups. Teen childbearing rose 3% in 2006, to 41.9 per 1000 females aged 15 to 19 years, the first increase after 14 years of steady decline. Birth rates rose 2% to 4% for women aged 20 to 44; rates for the youngest (10-14 years) and oldest (45-49) women were unchanged. Childbearing by unmarried women increased steeply in 2006 and set new historic highs. The cesarean-delivery rate rose by 3% in 2006 to 31.1% of all births; this figure has been up 50% over the last decade. Preterm and low birth weight rates also increased for 2006 to 12.8% and 8.3%, respectively. The 2005 infant mortality rate was 6.89 infant deaths per 1000 live births, not statistically higher than the 2004 level. Non-Hispanic black newborns continued to be more than twice as likely as non-Hispanic white and Hispanic infants to die in the first year of life in 2004. For all gender and race groups combined, expectation of life at birth reached a record high of 77.9 years in 2005. Age-adjusted death rates in the United States continue to decline. The crude death rate for children aged 1 to 19 years decreased significantly between 2000 and 2005. Of the 10 leading causes of death for children in 2005, only the death rate for cerebrovascular disease was up slightly from 2000, whereas accident and chronic lower respiratory disease death rates decreased. A large proportion of childhood deaths, however, continue to occur as a result of preventable injuries. PMID- 18381545 TI - Love, pain, and intensive care. PMID- 18381546 TI - Guidelines for early identification, screening, and clinical management of children with autism spectrum disorders. PMID- 18381547 TI - First, do no harm. PMID- 18381548 TI - A response to the expert committee's recommendations on the assessment, prevention, and treatment of child and adolescent overweight and obesity. PMID- 18381550 TI - Medical conditions affecting sports participation. AB - Children and adolescents with medical conditions present special issues with respect to participation in athletic activities. The pediatrician can play an important role in determining whether a child with a health condition should participate in certain sports by assessing the child's health status, suggesting appropriate equipment or modifications of sports to decrease the risk of injury, and educating the athlete, parent(s) or guardian, and coach regarding the risks of injury as they relate to the child's condition. This report updates a previous policy statement and provides information for pediatricians on sports participation for children and adolescents with medical conditions. PMID- 18381549 TI - Strength training by children and adolescents. AB - Pediatricians are often asked to give advice on the safety and efficacy of strength-training programs for children and adolescents. This statement, which is a revision of a previous American Academy of Pediatrics policy statement, defines relevant terminology and provides current information on risks and benefits of strength training for children and adolescents. PMID- 18381551 TI - Management of pediatric trauma. AB - Injury is the number 1 killer of children in the United States. In 2004, injury accounted for 59.5% of all deaths in children younger than 18 years. The financial burden to society of children who survive childhood injury with disability continues to be enormous. The entire process of managing childhood injury is complex and varies by region. Only the comprehensive cooperation of a broadly diverse group of people will have a significant effect on improving the care and outcome of injured children. This statement has been endorsed by the American Association of Critical-Care Nurses, American College of Emergency Physicians, American College of Surgeons, American Pediatric Surgical Association, National Association of Children's Hospitals and Related Institutions, National Association of State EMS Officials, and Society of Critical Care Medicine. PMID- 18381552 TI - Financing graduate medical education to meet the needs of children and the future pediatrician workforce. AB - This policy statement articulates the positions of the American Academy of Pediatrics on graduate medical education and the associated costs and funding mechanisms. It reaffirms the policy of the American Academy of Pediatrics that graduate medical education is a public good and is an essential part of maintaining a high-quality physician workforce. The American Academy of Pediatrics advocates for lifelong learning across the continuum of medical education. This policy statement focuses on the financing of one component of this continuum, namely residency education. The statement calls on federal and state governments to continue their support of residency education and advocates for stable means of funding such as the establishment of an all-payer graduate medical education trust fund. It further proposes a portable authorization system that would allocate graduate medical education funds for direct medical education costs to accredited residency programs on the basis of the selection of the program by qualified student or residents. This system allows the funding to follow the residents to their program. Recognizing the critical workforce needs of many pediatric medical subspecialties, pediatric surgical specialties, and other pediatric specialty disciplines, this statement maintains that subspecialty fellowship training and general pediatrics research fellowship training should receive adequate support from the graduate medical education financing system, including funding from the National Institutes of Health and other federal agencies, as appropriate. Furthermore, residency education that is provided in freestanding children's hospitals should receive a level of support equivalent to that of other teaching hospitals. The financing of graduate medical education is an important and effective tool to ensure that the future pediatrician workforce can provide optimal heath care for infants, children, adolescents, and young adults. PMID- 18381553 TI - Low birth weight and retinal vascular caliber in young children. PMID- 18381554 TI - Cardiac troponins in pediatric myocarditis. PMID- 18381555 TI - Quality of amino acid solutions for preterm infants. PMID- 18381556 TI - Bilirubin-binding capacity in low birthweight premature infants. PMID- 18381557 TI - Severe respiratory syncytial virus infection in term infants with genetic or other underlying disorders. PMID- 18381558 TI - Methadone maintenance and breastfeeding in the neonatal period. PMID- 18381559 TI - Corticosteroid therapy for Henoch Schonlein purpura. PMID- 18381560 TI - Bilirubin-binding capacity in premature infants. PMID- 18381561 TI - Youth caregivers: unrecognized providers of care. PMID- 18381562 TI - Refining insights into high-affinity drug binding to the human ether-a-go-go related gene potassium channel. AB - hERG (human ether-a-go-go-related gene) potassium (K(+)) channels play a crucial role in electrophysiological activity in the heart, exerting a profound influence on ventricular action potential repolarization and on the duration of the QT interval of the electrocardiogram. hERG channels are strongly implicated in the acquired form of long QT syndrome in that they exhibit a unique susceptibility to pharmacological inhibition by therapeutically and chemically diverse drugs. Investigations over a number of years provide compelling evidence that a comparatively large inner cavity and the presence of particular aromatic amino acid residues (Tyr652 and Phe656) on the inner (S6) helices of the channel are important features that allow hERG to accommodate and bind disparate drugs. However, whereas functional hERG channels are composed of four identical subunits, blocking molecules may not interact equally with aromatic residues from each of the four subunits. In this issue of Molecular Pharmacology, Myokai et al. (p. 1643) report for the first time the use of tandem dimers incorporating mutations to Tyr652 and Phe656 to elucidate asymmetric binding of the high affinity hERG inhibitor cisapride. Not only has this approach provided increased information on spatial arrangements involved in cisapride binding to the channel, but it offers a powerful means of refining the wider understanding of hERG channel structure-function in relation to drug binding. PMID- 18381563 TI - Roles of accessory subunits in alpha4beta2(*) nicotinic receptors. AB - Accessory subunits in heteromeric nicotinic receptors (AChRs) do not take part in forming ACh binding sites. alpha5 and beta3 subunits can function only as accessory subunits. We show that both alpha5 and beta3 efficiently assemble in human alpha4beta2(*) AChRs expressed in permanently transfected human embryonic kidney (HEK) cell lines. Only (alpha4beta2)(2)alpha5, not (alpha4beta2)(2)beta3 AChRs, have been detected in brain. The alpha4beta2alpha5 line expressed 40% more AChRs than the parent alpha4beta2 line and was equally sensitive to up-regulation by nicotine. The alpha4beta2beta3 line expressed 25-fold more AChRs than the parental line and could not be further up-regulated by nicotine. Relative sensitivity to activation by ACh depends on the accessory subunit, beta2 conferring the greatest sensitivity, alpha5 less, and beta3 and alpha4 much less. Accessory subunits form binding sites for positive allosteric modulators, as illustrated by the observation that alpha5 conferred high sensitivity to galanthamine. In the presence of alpha5 or beta3, stable, partially degraded, dead end intermediates accumulated within the cells. These may have the form alpha5alpha4beta2alpha5. The efficiency with which alpha5 and beta3 assemble with alpha4 and beta2 and the necessity of avoiding formation of potentially toxic intermediates may explain why alpha5 and beta3 seem to be transcribed at low levels in brain. Autosomal dominant nocturnal frontal lobe epilepsy can be caused by the alpha4 mutation S247F. This mutant did not produce functional AChRs unless cells were cotransfected with alpha5, beta3, or alpha6 to replace alpha4 as accessory subunit. PMID- 18381564 TI - Identification of regions required for apical membrane localization of human multidrug resistance protein 2. AB - Multidrug resistance proteins MRP1 and MRP2 transport a wide range of endo- and xenobiotics. However, with the exception of certain parts of the brain, MRP1 traffics to basolateral membranes of polarized cells, whereas MRP2 is apical in location and thus it is particularly important for systemic elimination of such compounds. Different regions of MRP1 and MRP2 seem to target them to their respective membrane locations. In addition to two "core" membrane spanning domains (MSDs) characteristic of ATP-binding cassette transporters, MRP1 and MRP2 have a third NH2-terminal MSD (MSD0), which is not required for basolateral targeting of MRP1, or for transport of at least some substrates. Here, we demonstrate that all elements necessary for apical targeting of MRP2 reside in MSD0 and the adjacent cytoplasmic loop (CL) 3. Furthermore, we show that this region of MRP2 can target the core of MRP1 to an exclusively apical location. Within MRP2 CL3, we identified a lysine-rich element that is essential for apical targeting. When introduced into MRP1, this element alone is sufficient to result in partial apical localization. However, exclusive targeting to the apical membrane seems to require the integrity of the entire region encompassing MSD0 and CL3 of MRP2. Because CL3 of MRP1 is critical for binding, transport, or both of several compounds, we also examined the function of hybrids containing all, or portions of MRP2 MSD0 and CL3. Our results indicate that CL3 is important for interaction with both the glutathione and glucuronide conjugates tested, but that different regions may be involved. PMID- 18381565 TI - Organic anion transporter 3 (oat3/slc22a8) interacts with carboxyfluoroquinolones, and deletion increases systemic exposure to ciprofloxacin. AB - Carboxyfluoroquinolones, such as ciprofloxacin, are used for the treatment of numerous infectious diseases. Renal secretion is a major determinant of their systemic and urinary concentration, but the specific transporters involved are virtually unknown. In vivo studies implicate the organic anion transporter (OAT) family as a pivotal component of carboxyfluoroquinolone renal secretion. Therefore, this study identified the specific renal basolateral OAT(s) involved, thereby highlighting potential sources of carboxyfluoroquinolone-drug interactions and variable efficacy. Two heterologous expression systems, Xenopus laevis oocytes and cell monolayers, were used to determine the roles of murine and human renal basolateral mOat1/hOAT1 and mOat3/hOAT3. Ciprofloxacin was transported by mOat3 in both systems (K(m) value, 70 +/- 6 microM) and demonstrated no interaction with mOat1 or hOAT1. Furthermore, ciprofloxacin, norfloxacin, ofloxacin, and gatifloxacin exhibited concentration-dependent inhibition of transport on mOat3 in cells with inhibition constants of 198 +/- 39, 558 +/- 75, 745 +/- 165, and 941 +/- 232 microM, respectively. Ciprofloxacin and gatifloxacin also inhibited hOAT3. Thereafter, in vivo elimination of ciprofloxacin was assessed in wild-type and Oat3 null mice [Oat3-/-]. Oat3-/- mice exhibited significantly elevated plasma levels of ciprofloxacin at clinically relevant concentrations (P < 0.05, male mice; P < 0.01, female mice). Oat3-/- mice also demonstrated a reduced volume of distribution (27%, P < 0.01, male mice; 14%, P < 0.01, female mice) and increased area under the concentration time curve (25%, P < 0.05, male mice; 33%, P < 0.01, female mice). Female Oat3-/- mice had a 35% (P < 0.01) reduction in total clearance of ciprofloxacin relative to wild type. In addition, putative ciprofloxacin metabolites were significantly elevated in Oat3-/- mice. The present findings indicate that polymorphisms of and drug interactions on hOAT3 may influence carboxyfluoroquinolone efficacy, especially in urinary tract infections. PMID- 18381566 TI - A 50-year-old man with chronic low back pain. AB - Mr S, a 50-year-old man, has long-standing low back pain. His pain began more than 20 years earlier with a lumbar disk herniation and has persisted despite diskectomy. He has undergone numerous treatments, but he remains disabled with ongoing pain. His treatment course is used to frame the epidemiology and pathophysiology underlying acute and chronic lumbosacral and radicular pain. The roles of neuropathic pain medications, chronic opioid therapy, physical therapy, spinal manipulation, and multidisciplinary pain treatment programs are reviewed. The indications for and outcomes associated with interventional pain treatments, including epidural steroid injection, facet blocks and radiofrequency treatment for facet-related pain, intradiskal electrothermal therapy, spinal cord stimulation, and intrathecal drug delivery, are discussed. Clinicians are given an evidence-based approach to using available treatment options for low back pain. PMID- 18381568 TI - Endometrial inhibin/activin beta-B subunit expression is related to decidualization and is reduced in tubal ectopic pregnancy. AB - CONTEXT: Ectopic pregnancy is common but remains difficult to diagnose accurately. There is no serum test to differentiate ectopic from intrauterine gestation. OBJECTIVE: Our objective was to investigate differential gene expression in decidualized endometrium of ectopic pregnancy. DESIGN: Tissue and serum analysis informed by microarray study was performed. SETTING: The study was performed at a large United Kingdom teaching hospital. PATIENTS OR OTHER PARTICIPANTS: Women undergoing surgical termination of pregnancy (n = 8), evacuation of uterus for miscarriage (n = 6), and surgery for tubal ectopic pregnancy (n = 11) were included in the study. Endometrium was collected from normally cycling women undergoing hysterectomy. INTERVENTIONS: Decidualized endometrium was subjected to microarray analysis, morphological assessment, and immunohistochemistry. Endometrial stromal fibroblasts were cultured in the presence of decidualizing stimuli. MAIN OUTCOME MEASURES: Differential expression of potentially secreted molecules was calculated. RESULTS: Inhibin/activin beta-B expression was lower in decidualized endometrium from ectopic pregnancies when compared with that of ongoing pregnancies (P < 0.01) or miscarriages (P < 0.01). The localization of the beta-B subunit was more marked in decidualized than nondecidualized stroma. Decidualization of stromal fibroblasts in vitro was associated with increased beta-B expression (P < 0.05). Endometrial stroma of ectopic pregnancies was less decidualized morphologically (P < 0.05), with lower prolactin (P < 0.01) and IGF binding protein-1 (P < 0.005) expression. Serum activin B was lower in ectopic pregnancies (P < 0.005) than in intrauterine pregnancies, whereas there was no difference in progesterone concentrations. CONCLUSIONS: Despite similar concentrations of progesterone, the endometrium of ectopic pregnancies is less decidualized than intrauterine pregnancies. Expression of the beta-B subunit is related to decidualization and can be detected in the circulation as activin B. Serum activin B concentrations are lower in ectopic pregnancy. PMID- 18381569 TI - Effects of an early postnatal treatment of hypogonadotropic hypogonadism with a continuous subcutaneous infusion of recombinant follicle-stimulating hormone and luteinizing hormone. AB - BACKGROUND: The neonatal-midinfancy surge in pulsatile gonadotropin secretion is attributable to an increase in GnRH pulse amplitude and is associated with a rapid expansion of Leydig and Sertoli cell populations with concomitant surges in testosterone, inhibin, and anti-Mullerian hormone production as well as an increase in testicular volume. Boys with congenital hypogonadotropic hypogonadism (HH) do not activate these processes. A potential cause for azoospermia and infertility in adult life is deficient proliferation of immature Sertoli cells before and during puberty due to the absence of FSH. OBJECTIVE: The objective of the study was to investigate whether early postnatal continuous sc infusion of gonadotropins could mimic the physiological growth of testes and to evaluate responses of the Leydig and Sertoli cells to early gonadotropin replacement. DESIGN AND METHODS: Two neonates (P1 with hypotuitarism and P2 with HH) with micropenis and microorchidism were treated for 6 months with high doses of recombinant LH and FSH (a gift of Luveris and Gonal-F from Serono, Lyon, France) delivered sc with an insulin pump. RESULTS: Gonadotropin continuous sc infusion increased mean serum LH and FSH to normal or supranormal levels. Mean testosterone increased from undetectable levels to 7.6 and 5.2 nmol/liter, respectively, in P1 and P2. Inhibin B and anti-Mullerian hormone increased to normal levels. Mean testicular volume increased from 0.45 to 0.57 ml at birth to 2.10 ml at 7 months. Stretched penile length increased from 8 to 30 mm (P1) and 12 to 48 mm (P2). CONCLUSIONS: The present regimen induced physiological postnatal testes growth and high-normal activation of Leydig and Sertoli cells. PMID- 18381567 TI - Efficacy and safety of pyridoxal 5'-phosphate (MC-1) in high-risk patients undergoing coronary artery bypass graft surgery: the MEND-CABG II randomized clinical trial. AB - CONTEXT: Coronary artery bypass graft (CABG) surgery is frequently performed and effective; however, perioperative complications related to ischemia-reperfusion injury, including myocardial infarction (MI), remain common and result in significant morbidity and mortality. MC-1, a naturally occurring pyridoxine metabolite and purinergic receptor antagonist, prevents cellular calcium overload and may reduce ischemia-reperfusion injury. Phase 2 trial data suggest that MC-1 may reduce death or MI in high-risk patients undergoing CABG surgery. OBJECTIVE: To assess the efficacy and safety of MC-1 administered immediately before and for 30 days after surgery in patients undergoing CABG surgery. DESIGN, SETTING, AND PARTICIPANTS: The MC-1 to Eliminate Necrosis and Damage in Coronary Artery Bypass Graft Surgery II Trial, a phase 3, multicenter, randomized, double-blind, placebo controlled trial, with 3023 intermediate- to high-risk patients undergoing CABG surgery with cardiopulmonary bypass enrolled between October 2006 and September 2007 at 130 sites in Canada, the United States, and Germany. INTERVENTIONS: Patients received either MC-1, 250 mg/d (n = 1519), or matching placebo (n = 1504) immediately before and for 30 days after CABG surgery. MAIN OUTCOME MEASURES: The primary efficacy outcome was cardiovascular death or nonfatal MI, defined as a creatine kinase (CK) MB fraction of at least 100 ng/mL or new Q waves through postoperative day 30. RESULTS: The primary efficacy outcome occurred in 140 of 1510 patients (9.3%) in the MC-1 group and 133 of 1486 patients (9.0%) in the placebo group (risk ratio, 1.04; 95% confidence interval, 0.83-1.30; P = .76). All-cause mortality was higher among patients assigned to MC 1 than placebo at 4 days (1.0% vs 0.3%; P = .03) but was similar at 30 days (1.9% vs 1.5%; P = .44). There was no difference in the 8- to 24-hour CK-MB area under the curve between the MC-1 and placebo groups (median, 270 [interquartile range, 175-492] vs 268 [interquartile range, 170-456] hours x ng/mL; P = .11). CONCLUSION: In this population of intermediate- to high-risk patients undergoing CABG surgery, MC-1 did not reduce the composite of cardiovascular death or nonfatal MI. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00402506 PMID- 18381570 TI - BRAFV600E mutation is associated with preferential sensitivity to mitogen activated protein kinase kinase inhibition in thyroid cancer cell lines. AB - CONTEXT: Mutually exclusive mutations of RET, RAS, or BRAF are present in about 70% of papillary thyroid carcinomas, whereas only the latter two are seen in poorly differentiated and anaplastic cancers. Although the signal output common to these oncoproteins is ERK, a recent report showed that only BRAF mutations consistently predicted responsiveness to MAPK kinase (MEK) inhibitors. OBJECTIVES: Here we investigated whether sensitivity to MEK inhibition was determined by oncogene status in 13 human thyroid cancer cell lines: four with BRAF mutations, four RAS, one RET/PTC1, and four wild type. RESULTS: Growth of BRAF (+) cells was inhibited by the MEK antagonist PD0325901 with an IC(50) of less than 5 nm. By contrast, RAS, RET/PTC1, or wild-type cells had IC(50) of 4 nm to greater than 1000 nm. Sensitivity was not predicted by coexisting mutations in PIK3CA or by PTEN status. Similar effects were obtained with the MEK inhibitor AZD6244. PD0325901 induced a sustained G1/S arrest in BRAF (+) but not BRAF (-) lines. PD0325901 was equipotent at inhibiting pERK1/2 after 2 h, regardless of genetic background, but pERK rebounded at 24 h in most lines. MEK inhibitor resistance was associated with partial refractoriness of pERK to further inhibition by the compounds. AZD6244 was more potent at inhibiting growth of NPA (BRAF +) than Cal62 (KRAS +) xenografts. CONCLUSION: Thyroid cancers with BRAF mutation are preferentially sensitive to MEK inhibitors, whereas tumors with other MEK-ERK effector pathway gene mutations have variable responses, either because they are only partially dependent on ERK and/or because feedback responses elicit partial refractoriness to MEK inhibition. PMID- 18381571 TI - Effects of denosumab on bone mineral density and bone turnover in postmenopausal women. AB - CONTEXT: Denosumab is an investigational fully human monoclonal antibody against receptor activator of nuclear factor-kappaB ligand, a mediator of osteoclastogenesis and osteoclast survival. OBJECTIVE: This study evaluated the ability of denosumab to increase bone mineral density (BMD) and decrease bone turnover markers (BTMs) in early and later postmenopausal women with low BMD. DESIGN AND SETTING: This 2-yr randomized, double-blind, placebo-controlled study was conducted in North America. PARTICIPANTS: Subjects included 332 postmenopausal women with lumbar spine BMD T-scores between -1.0 and -2.5. INTERVENTIONS: SUBJECTS were randomly assigned to receive denosumab sc, 60 mg every 6 months, or placebo. Randomization was stratified by time since onset of menopause (< or =5 yr or > 5 yr). MAIN OUTCOME MEASURES: The primary end point was the percent change in lumbar spine BMD by dual-energy x-ray absorptiometry at 24 months. Additional end points were percent change in volumetric BMD of the distal radius by quantitative computed tomography; percent change in BMD by dual energy x-ray absorptiometry for the total hip, one-third radius, and total body; hip structural analysis; percent change in BTMs; and safety. RESULTS: Denosumab significantly increased lumbar spine BMD, compared with placebo at 24 months (6.5 vs. -0.6%; P<0.0001) with similar results for both strata. Denosumab also produced significant increases in BMD at the total hip, one-third radius, and total body (P < 0.0001 vs. placebo); increased distal radius volumetric BMD (P < 0.01); improved hip structural analysis parameters; and significantly suppressed serum C-telopeptide, tartrate-resistant acid phosphatase-5b, and intact N terminal propeptide of type 1 procollagen. The overall incidence of adverse events was similar between both study groups. CONCLUSIONS: Twice-yearly denosumab increased BMD and decreased BTMs in early and later postmenopausal women. PMID- 18381572 TI - The role of the aryl hydrocarbon receptor-interacting protein gene in familial and sporadic pituitary adenomas. AB - CONTEXT: Mutations have been identified in the aryl hydrocarbon receptor interacting protein (AIP) gene in familial isolated pituitary adenomas (FIPA). It is not clear, however, how this molecular chaperone is involved in tumorigenesis. OBJECTIVE: AIP sequence changes and expression were studied in FIPA and sporadic adenomas. The function of normal and mutated AIP molecules was studied on cell proliferation and protein-protein interaction. Cellular and ultrastructural AIP localization was determined in pituitary cells. PATIENTS: Twenty-six FIPA kindreds and 85 sporadic pituitary adenoma patients were included in the study. RESULTS: Nine families harbored AIP mutations. Overexpression of wild-type AIP in TIG3 and HEK293 human fibroblast and GH3 pituitary cell lines dramatically reduced cell proliferation, whereas mutant AIP lost this ability. All the mutations led to a disruption of the protein-protein interaction between AIP and phosphodiesterase-4A5. In normal pituitary, AIP colocalizes exclusively with GH and prolactin, and it is found in association with the secretory vesicle, as shown by double-immunofluorescence and electron microscopy staining. In sporadic pituitary adenomas, however, AIP is expressed in all tumor types. In addition, whereas AIP is expressed in the secretory vesicle in GH-secreting tumors, similar to normal GH-secreting cells, in lactotroph, corticotroph, and nonfunctioning adenomas, it is localized to the cytoplasm and not in the secretory vesicles. CONCLUSIONS: Our functional evaluation of AIP mutations is consistent with a tumor-suppressor role for AIP and its involvement in familial acromegaly. The abnormal expression and subcellular localization of AIP in sporadic pituitary adenomas indicate deranged regulation of this protein during tumorigenesis. PMID- 18381573 TI - Pharmacodynamics of growth hormone abuse biomarkers and the influence of gender and testosterone: a randomized double-blind placebo-controlled study in young recreational athletes. AB - CONTEXT: IGF axis proteins and collagen peptides are promising markers of GH abuse. OBJECTIVE: Our objective was to investigate whether responses of serum IGF axis and collagen markers to GH differ between men and women, and are influenced by testosterone (T). DESIGN: This was a randomized, double-blind, placebo controlled study of 8-wk treatment followed by 6-wk washout. SETTING: The study was performed at a clinical research facility. PARTICIPANTS: A total of 96 recreationally trained healthy athletes (63 men, 33 women), aged 18-40 yr, were studied. INTERVENTION: All subjects received GH (2 mg/d sc) or placebo for 8 wk; men also received T (250 mg/wk im) or placebo for 5 wk. MAIN OUTCOME MEASURES: Serum IGF axis proteins (IGF-I, IGF binding protein-3, and acid labile subunit) and collagen peptides (N-terminal propeptide of type I procollagen, C-terminal telopeptide of type I collagen, and N-terminal propeptide of type III procollagen) were measured. RESULTS: GH induced significant increases in IGF axis and collagen markers that were greater in men than women (P < 0.001). Of the IGF axis markers, IGF-I showed the greatest increase. The relative incremental responses of the collagen markers in general were greater than the IGF markers, especially for PIIINP. The collagen markers increased and decreased more slowly with most remaining elevated (P < 0.01) after 6 wk, in comparison to IGF markers, which returned to baseline within 1 wk. Addition of T to GH amplified the response of PIIINP by more than 1.5-fold but did not affect any other marker. T alone did not affect IGF axis markers but modestly increased collagen markers. CONCLUSIONS: These markers of GH abuse are less responsive in women. The increases in collagen markers have a different time course to the IGF markers and extend the window of detection in both sexes. The response of PIIINP is increased by coadministration of T. PMID- 18381574 TI - Expression of six transmembrane protein of prostate 2 in human adipose tissue associates with adiposity and insulin resistance. AB - CONTEXT: Six transmembrane protein of prostate 2 (STAMP2) is a counterregulator of adipose inflammation and insulin resistance in mice. Our hypothesis was that STAMP2 could be involved in human obesity and insulin resistance. OBJECTIVE: The objective of the study was to elucidate the role of adipose STAMP2 expression in human obesity and insulin resistance. DESIGN: The design was to quantify STAMP2 in human abdominal sc and omental white adipose tissue (WAT), isolated adipocytes, and stroma and in vitro differentiated preadipocytes and relate levels of STAMP2 in sc WAT to clinical and adipocyte phenotypes involved in insulin resistance. PARTICIPANTS: Nonobese and obese women and men (n = 236) recruited from an obesity clinic or through local advertisement. MAIN OUTCOME MEASUREMENT: Clinical measures included body mass index, body fat, total adiponectin, and homeostasis model assessment as measure of overall insulin resistance. In adipocytes we determined cell size, sensitivity of lipolysis and lipogenesis to insulin, adiponectin secretion, and inflammatory gene expression. RESULTS: STAMP2 levels in sc and visceral WAT and adipocytes were increased in obesity (P = 0.0008-0.05) but not influenced by weight loss. Increased WAT STAMP2 levels associated with a high amount of body fat (P = 0.04), high homeostasis model assessment (P = 0.01), and large adipocytes (P = 0.02). Subjects with high STAMP2 levels displayed reduced sensitivity of adipocyte lipogenesis (P = 0.04) and lipolysis (P = 0.03) to insulin but had normal adiponectin levels. WAT STAMP2 levels correlated with expression of the macrophage marker CD68 (P = 0.0006). CONCLUSION: Human WAT STAMP2 associates with obesity and insulin resistance independently of adiponectin, but the role of STAMP2 in obesity and its complications seems different from that in mice. PMID- 18381575 TI - Size, number, and distribution of thyroid nodules and the risk of malignancy in radiation-exposed patients who underwent surgery. AB - CONTEXT: The chance that a thyroid nodule is malignant is higher when there is a history of childhood radiation exposure. OBJECTIVE: The objective of the study was to determine how the size of a thyroid nodule, the number of nodules, and the distribution of nodules influence the risk of cancer in irradiated patients. PATIENTS: From a cohort of 4296 radiation-exposed people, we studied the 1059 that underwent thyroid surgery. DESIGN AND OUTCOMES: We studied the association between the size, number, distribution, and rank order of thyroid nodules and the chance of malignancy. RESULTS: There were 612 malignant nodules in 358 patients and 2037 benign ones in 930 patients. There was no change in the risk that a nodule was malignant with increasing size (odds ratio 0.91/cm, P = 0.11) among the 1709 nodules that were 0.5 cm or greater. A solitary nodule had a similar likelihood of being malignant as a nodule that was one of several (18.8 vs. 17.3%), whereas patients with multiple nodules were more likely to have thyroid cancer than those with solitary nodules [30.7 vs. 18.7%; risk ratio 1.64 (1.27 2.13)]. Aspirating only the largest nodule would have missed 111 of the cancers (42%), whereas aspirating the two largest nodules would have missed 45 of the cases (17%), although none would have been 10 mm or greater. CONCLUSIONS: In radiation-exposed patients, the following conclusions were made: 1) the likelihood that a nodule is malignant is independent of nodule number and size; 2) the likelihood of cancer is increased if more than one nodule is present; 3) evaluating the two largest nodules by fine-needle aspiration would have resulted in a significant number of cases being missed but none with large cancers; and 4) more than half of the patients with thyroid cancer had multifocal tumors. PMID- 18381576 TI - Expression and function of Cbfa-1/Runx2 in thyroid papillary carcinoma cells. AB - CONTEXT: Development of calcifying foci is a common finding in human thyroid papillary carcinoma, but its mechanisms remain unknown. OBJECTIVE: We therefore investigated whether osteocalcin and/or Cbfa-1 genes are expressed in malignant thyroid epithelial cells. We also studied the effects of Cbfa-1 on the expression of osteoblast-specific and thyrotropin receptor genes in thyrocytes. RESULTS: The human thyroid papillary carcinoma cell line BHP18-21 expresses bone-type osteocalcin mRNA at higher levels than in MG63 osteosarcoma cells. Northern blot analysis and EMSA using nuclear extracts from BHP18-21 cells and FRTL-5 cells demonstrated that cells of thyroid epithelial origin expressed Cbfa-1/Runx2, the main transcription factor for the expression of osteocalcin. When we transfected pcDNA3.1-human Cbfa-1 into FRTL-5 cells, Cbfa-1 increased the gene expression of alkaline phosphatase, type I collagen, and osteocalcin but suppressed the expression of thyrotropin receptor. We then stained the calcified regions of human papillary thyroid carcinoma tissues with antiosteocalcin antibody and found that malignant cells, as well as follicular epithelial cells, were immunopositive for osteocalcin. Northern blot analysis revealed that the Cbfa-1/Runx2 gene was strongly expressed in tissues from four cases of surgically resected papillary carcinoma. CONCLUSIONS: Thyrocytes share characteristics with osteoblasts. Cbfa-1 may play a role in calcification processes in human thyroid papillary carcinoma tissues. PMID- 18381578 TI - Serum ghrelin levels are increased in hypothyroid patients and become normalized by L-thyroxine treatment. AB - CONTEXT: An interaction between ghrelin, which is implicated in the regulation of short- and long-term energy balance, and thyroid function has been reported in hyperthyroidism in which ghrelin levels are reversibly suppressed. We measured serum ghrelin levels and metabolic indices in hypothyroid patients before and after L-thyroxine replacement. PATIENTS AND METHODS: Eleven patients were examined twice: 1) in the hypothyroid state and 2) after at least 2 months of euthyroidism. Ten healthy subjects served as a control group. Ghrelin was measured in conjunction with indirect calorimetry and a hyperinsulinemic euglycemic clamp. RESULTS: Serum ghrelin levels were increased by 32% under basal conditions in the hypothyroid state (PRE) as compared with posttreatment (POST) (picograms per milliliter): 976.4 +/- 80.8 vs. 736.8 +/- 67.1 (P < 0.001). This difference prevailed during the clamp, but a decline was observed in both states: 641.4 +/- 82.2 vs. 444.3 +/- 66.8 microg/ml (P = 0.005). The hypothyroid state was associated with decreased resting energy expenditure, increased respiratory quotient, and insulin resistance. Serum ghrelin levels as well as the metabolic aberrations became normalized after L-thyroxine replacement as compared with the control subjects. CONCLUSION: Serum ghrelin levels are reversibly increased in hypothyroid patients. It remains to be investigated whether this represents a direct effect of iodothyronines on ghrelin secretion or clearance or a compensatory response to the abnormal energy metabolism in hypothyroid patients. PMID- 18381577 TI - Reciprocal relation between marrow adiposity and the amount of bone in the axial and appendicular skeleton of young adults. AB - BACKGROUND: Studies in the elderly suggest a reciprocal relation between increased marrow adiposity and bone loss, supporting basic research data indicating that osteoblasts and adipocytes share a common progenitor cell. However, whether this relation represents a preferential differentiation of stromal cells from osteoblasts to adipocytes or whether a passive accumulation of fat as bone is lost and marrow space increases with aging is unknown. To address this question and avoid the confounding effect of bone loss, we examined teenagers and young adults. METHODS: Using computed tomography, we obtained measurements of bone density and cross-sectional area of the lumbar vertebral bodies and cortical bone area, cross-sectional area, marrow canal area, and fat density in the marrow of the femurs in 255 sexually mature subjects (126 females, 129 males; 15-24.9 yr of age). Additionally, values for total body fat were obtained with dual-energy x-ray absorptiometry. RESULTS: Regardless of gender, reciprocal relations were found between fat density and measures of vertebral bone density and femoral cortical bone area (r = 0.19-0.39; all P values < or = .03). In contrast, there was no relation between marrow canal area and cortical bone area in the femurs, neither between fat density and the cross-sectional dimensions of the bones. We also found no relation between anthropometric or dual energy x-ray absorptiometry fat values and measures for marrow fat density. CONCLUSIONS: Our results indicate an inverse relation between bone marrow adiposity and the amount of bone in the axial and appendicular skeleton and support the notion of a common progenitor cell capable of mutually exclusive differentiation into the cell lineages responsible for bone and fat formation. PMID- 18381579 TI - Inhibition of CYP21A2 enzyme activity caused by novel missense mutations identified in Brazilian and Scandinavian patients. AB - BACKGROUND: Most patients with 21-hydroxylase deficiency carry CYP21A1P-derived mutations, but an increasing number of novel and rare mutations have been reported in disease-causing alleles. OBJECTIVE: Functional effects of three novel (p.G56R, p.L107R, p.L142P) and one recurrent (p.R408C) CYP21A2 mutations were investigated. The degree of enzyme impairment caused by p.H62L alone or combined to p.P453S was also analyzed. DESIGN: The study included 10 Brazilian and two Scandinavian patients. To determine the deleterious role of each mutant protein, in vitro assays were performed in transiently transfected COS-1 cells. For a correct genotype-phenotype correlation, the enzymatic activities were evaluated toward the two natural substrates, 17-hydroxyprogesterone and progesterone. RESULTS: Low levels of residual activities obtained for p.G56R, p.L107R, p.L142P, and p.R408C mutants classified them as classical congenital adrenal hyperplasia mutations, whereas the p.H62L showed an activity within the range of nonclassical mutations. Apparent kinetic constants for p.H62L confirmed the nonclassical classification as the substrate binding capacity was within the same magnitude for mutant and normal enzymes. A synergistic effect was observed for the allele bearing the p.H62L+p.P453S combination because it caused a significant reduction in the enzymatic activity. CONCLUSIONS: We describe the functional analysis of five rare missense mutations identified in Brazilian and Scandinavian patients. The p.G56R, p.L107R, and p.L142P are reported for the first time. Most probably these novel mutations are closer to null than the p.I172N, but for the p.G56R, that might not be the case, and the p.H62L is definitely a nonclassical mutation. PMID- 18381580 TI - Retinol binding protein-4 elevation is associated with serum thyroid-stimulating hormone level independently of obesity in elderly subjects with normal glucose tolerance. AB - CONTEXT: Elevated levels of retinol binding protein-4 (RBP4) are positively correlated with insulin resistance, obesity, diabetes mellitus, and cardiovascular disease (CVD). Subclinical hypothyroidism (SCH) has also been associated with CVD; however, the factors linking SCH to CVD are not clear. OBJECTIVE: The objective of the study was to evaluate risk factors for CVD in elderly patients grouped according to thyroid function. DESIGN: 217 subjects (> or =65 yr old) were randomly selected from a population and allocated to a euthyroid group (n = 177) and an SCH group (n = 40) on the basis of plasma concentrations of TSH and free T(4). We included subjects with normal glucose tolerance by a 75-g oral glucose tolerance test and subjects with impaired fasting glucose. We measured anthropometric parameters, levels of fasting glucose and insulin, hemoglobin A(1c), adiponectin, RBP4, lipid profiles, total body fat content, and the area of sc and visceral fat. RESULTS: The SCH group had higher RBP4 levels than the euthyroid group, irrespective of body mass index and fat content. Subcutaneous and visceral fat areas and total body fat percentage did not differ between groups and were not correlated with RBP4 level. Other CVD risk factors did not differ between groups. RBP4 level was positively correlated with TSH level (r = 0.241, P = 0.001) after adjustment for age, sex, and body mass index. CONCLUSIONS: Plasma RBP4 levels were associated with SCH independent of obesity in elderly subjects with normal glucose tolerance, indicating that RBP4 level could be used as an index of CVD risk in SCH. PMID- 18381582 TI - Concentration of anti-Mullerian hormone and inhibin-B in relation to steroids and age in follicular fluid from small antral human follicles. AB - CONTEXT: Ovaries surgically removed for fertility preservation served as a source of follicle fluid from human small antral follicles. OBJECTIVE: The objective of the study was to measure intrafollicular concentrations of anti-Mullerian Hormone (AMH), inhibin-B, progesterone, androstenedione, testosterone, estradiol, and IGF binding protein-4. SETTING: The study was conducted at a university hospital. PATIENTS: Patients included 43 women having one ovary removed prior to receiving gonadotoxic treatment due to malignant disease. INTERVENTIONS: Fluid from 100 follicles (diameter of 3-9 mm) were included. MAIN OUTCOME MEASURES: Intrafollicular concentrations of the measured hormones, their possible intercorrelation, and correlation with age were measured. RESULTS: Concentrations of AMH were unrelated to follicular fluid concentrations of androstenedione and testosterone. There was a significant negative correlation between estradiol, inhibin-B, progesterone, and AMH. In four age groups spanning 11-37 yr, levels of AMH, estradiol, androstenedione, testosterone and inhibin-B remained constant, whereas progesterone showed significant variations. IGF binding protein-4 was unrelated to any other measured hormone. CONCLUSIONS: This study was unable to confirm a stimulatory effect of androgens on AMH secretion but did enforce a close intimate correlation between AMH and estradiol expressions in the developing human follicle. The insignificant variation of the AMH concentration with age, even in prepubertal girls, suggests that AMH expression is unrelated to menstrual cycle FSH cyclicity. PMID- 18381581 TI - Growth hormone decreases visceral fat and improves cardiovascular risk markers in women with hypopituitarism: a randomized, placebo-controlled study. AB - CONTEXT: Data regarding gender-specific efficacy of GH on critical endpoints are lacking. There are no randomized, placebo-controlled studies of physiological GH therapy solely in women. OBJECTIVE: Our objective was to determine the effects of physiological GH replacement on cardiovascular risk markers and body composition in women with GH deficiency (GHD). DESIGN: This was a 6-month, randomized, placebo-controlled, double-blind study. SETTING: The study was conducted at the General Clinical Research Center. STUDY PARTICIPANTS: 43 women with GHD due to hypopituitarism were included in the study. INTERVENTION: Study participants were randomized to receive GH (goal mid-normal serum IGF-1) or placebo. MAIN OUTCOME MEASURES: Cardiovascular risk markers, including high-sensitivity C-reactive protein, tissue plasminogen activator, and body composition, including visceral adipose tissue by cross-sectional computed tomography, were measured. RESULTS: Mean daily GH dose was 0.67 mg. The mean IGF-1 sd score increased from -2.5 +/- 0.3 to -1.4 +/- 0.9 (GH) (P < 0.0001 vs. placebo). High-sensitivity C-reactive protein decreased by 38.2 +/- 9.6% (GH) vs.18.2 +/- 6.0% (placebo) (P = 0.03). Tissue plasminogen activator and total cholesterol decreased, and high-density lipoprotein increased. Homeostasis model assessment-insulin resistance and other markers were unchanged. Body fat decreased [-5.1 +/- 2.0 (GH) vs. 1.9 +/- 1.0% (placebo); P = 0.002] as did visceral adipose tissue [-9.0 +/- 5.9 (GH) vs. 4.3 +/- 2.7% (placebo); P = 0.03]. Change in IGF-1 level was inversely associated with percent change in visceral adipose tissue (r = -0.61; P = 0.002), total body fat (r = -0.69; P < 0.0001), and high-sensitivity C-reactive protein (r = -0.51; P = 0.003). CONCLUSIONS: Low-dose GH replacement in women with GHD decreased total and visceral adipose tissue and improved cardiovascular markers, with a relatively modest increase in IGF-1 levels and without worsening insulin resistance. PMID- 18381583 TI - No greater incidence or worsening of cardiac valve regurgitation with somatostatin analog treatment of acromegaly. AB - CONTEXT: Excess GH and IGF-I in acromegaly are associated with reduced life expectancy due to cardiovascular complications. OBJECTIVE: The objective of the study was to investigate the prevalence, incidence, and severity of cardiac valve regurgitation before and after somatostatin-analog treatment in acromegaly. DESIGN: This was a prospective, observer-blinded, multicenter, 12-month study. SETTING: The study was conducted at 33 specialist centers. PATIENTS: The study population consisted of 225 adult patients with acromegaly without significant cardiac valve abnormalities or prior valve-replacement surgery, matched for age, sex, and center/country/study. INTERVENTIONS: Interventions included initiation/continuation of lanreotide (n = 107) or octreotide treatment (n = 118), tailored for optimal disease control. MAIN OUTCOME MEASURES: Relative risk of new/worsening regurgitation in any valve at 12 months compared with baseline, was measured. RESULTS: At baseline, almost 80% of patients had some degree of cardiac valve regurgitation, although none was severe. The risk of developing new/worsening regurgitation in any valve at 12 months was nonsignificant and similar for the cohorts [adjusted odds ratio 0.86; 95% confidence interval (CI) 0.41-1.82; P = 0.694; relative risk 1.04; 95% CI 0.67-1.60; risk difference 0.01; 95% CI -0.13 to 0.16]. For 54% of patients, the severity of regurgitation stayed the same during the study. At baseline, significant valve regurgitation occurred in 18% of patients (lanreotide cohort) and 13% (octreotide cohort) and at 12 months in 18% of each cohort. CONCLUSIONS: The incidence of valve regurgitation did not change over 12 months of treatment with somatostatin analogs, and most cases were physiologic or mild in severity. There was no significant difference between somatostatin analogs in the risk of developing new/worsening valve regurgitation or significant regurgitation after 1 yr. PMID- 18381584 TI - Changing patterns in diagnosis and therapy of acromegaly over two decades. AB - BACKGROUND: The increased morbidity and mortality of acromegaly makes early diagnosis and therapy critical. However, whether the type of medical professional who first diagnoses acromegaly, the major complaint prompting medical attention, or the management paradigms used in the setting of novel medical therapies have changed over time has not been well explored. OBJECTIVES: Our objective was to identify the medical professional who first suspected acromegaly and the complaint prompting the diagnosis, and if these have changed. Additional goals were to assess the interval from symptom onset to diagnosis of acromegaly and to compare treatment trends over consecutive decades. DESIGN: This was a case-record retrospective study. SETTING: The study was performed in a neuroendocrine clinical center at a tertiary care center. SUBJECTS: A total of 100 patients (45 men and 55 women) with acromegaly referred from 1985-2005 was included in the study. RESULTS: Acral changes (24%) and headaches (20%) were most prevalent presenting symptoms prompting diagnosis. Eighteen percent reported no symptoms of acromegaly at diagnosis. The primary care physician most often initiated the evaluation (44%). Comorbidities were more prevalent in older patients (P = 0.001). The interval between symptom onset and diagnosis decreased, compared with previous reports. Radiation therapy was used less frequently in the decade after 1994 than in the prior (16 vs. 33%; P < 0.05). CONCLUSIONS: The primary care doctor plays the major role in diagnosis of acromegaly. The increased use of brain magnetic resonance imaging may contribute to the many incidentally discovered cases and to the shortened time interval to diagnosis. Presumably due to the availability of new medical therapies, the use of radiation therapy has decreased. PMID- 18381585 TI - Loss of the eukaryotic initiation factor 3f in melanoma. AB - Aberrant regulation of the translation initiation is known to contribute to tumorigenesis. eIF3 plays an important role in translation initiation. eIF3f is the p47 subunit of the eIF3 complex whose function in cancer is not clear. Initial studies from our group indicated that eIF3f expression is decreased in melanoma. Overexpression of eIF3f inhibits translation and induces apoptosis in melanoma cells. The eIF3f gene is located at chromosome region 11p15.4. Loss of 11p15.4 is a common event in many tumors including melanoma. In order to investigate the molecular mechanism of the decreased expression of eIF3f in melanoma, we performed loss of heterozygosity (LOH) analysis in 24 melanoma specimens using three microsatellite markers encompassing the eIF3f gene. We showed that the prevalence of LOH ranged from 75% to 92% in melanoma. We also performed eIF3f gene copy number analysis using quantitative real-time PCR to further confirm the specific allelic loss of the eIF3f gene in melanoma. We demonstrated a statistically significant decrease of the eIF3f gene copy number in melanoma compared with normal tissues with a tumor/normal ratio of 0.52. To further elucidate the somatic genetic alterations, we carried out mutation analysis covering the entire coding region and 5'UTR of the eIF3f gene in melanoma tissues and cell lines. Despite some polymorphisms, we did not find any mutations. Furthermore, immunohistochemistry analysis demonstrated that eIF3f protein expression is decreased in melanoma compared to benign nevi. These data provide new insight into the understanding of the molecular pathogenesis of eIF3f during melanoma tumorigenesis. PMID- 18381586 TI - Highly active ansamitocin derivatives: mutasynthesis using an AHBA-blocked mutant. PMID- 18381587 TI - Populations of p53 codon 270 CGT to TGT mutant cells in SKH-1 mouse skin tumors induced by simulated solar light. AB - The p53 codon 270 CGT to TGT mutation was investigated as a biomarker of sunlight induced mutagenesis and carcinogenesis. The relationship between tumor development and abundance of this hotspot mutation was analyzed in mouse skin tumors induced by chronic exposure to simulated solar light (SSL). The 24 tumors analyzed had similar growth kinetics, with an average doubling time of approximately 16.4 d. Levels of the p53 codon 270 mutation were quantified in the 24 mouse skin tumors using allele-specific competitive blocker-polymerase chain reaction (ACB-PCR). All tumors contained measurable amounts of the mutation. The p53 codon 270 CGT to TGT mutant fraction (MF) ranged from 2.29 x 10(-3) to 9.42 x 10(-2), with 3.26 x 10(-2) as the median. These p53 MF measurements are lower than expected for an initiating mutation involved in the development of tumors of monoclonal origin. There was no evidence of a correlation between p53 codon 270 MF and either tumor area or an estimate of tumor cell number. Thus, the data do not support the idea that p53 mutation accumulates linearly during tumor development. To investigate how p53 mutation was distributed within tumors, 19 needle biopsies from seven different tumors were analyzed by ACB-PCR. This analysis demonstrated that p53 codon 270 mutation is heterogeneously distributed within tumors. The long-term goal of this research is to combine morphological and p53 MF measurements from tissues corresponding to the various stages of tumor development, in order to derive mathematical models relating the p53 codon 270 mutation to the development of SSL-induced skin tumors. PMID- 18381588 TI - Evaluation of the statistical power for multiple tests: a case study. AB - It is challenging to estimate the statistical power when a complicated testing strategy is used to adjust for the type-I error for multiple comparisons in a clinical trial. In this paper, we use the Bonferroni Inequality to estimate the lower bound of the statistical power assuming that test statistics are approximately normally distributed and the correlation structure among test statistics is unknown or only partially known. The method was applied to the design of a clinical study for sample size and statistical power estimation. PMID- 18381589 TI - Resveratrol directly targets COX-2 to inhibit carcinogenesis. AB - Targeted molecular cancer therapies can potentially deliver treatment directly to a specific protein or gene to optimize efficacy and reduce adverse side effects often associated with traditional chemotherapy. Key oncoprotein and oncogene targets are rapidly being identified based on their expression, pathogenesis and clinical outcome. One such protein target is cyclooxygenase-2 (COX-2), which is highly expressed in various cancers. Research findings suggest that resveratrol (RSVL; 3,5,4'-trihydroxy-trans-stilbene) demonstrates nonselective COX-2 inhibition. We report herein that RSVL directly binds with COX-2 and this binding is absolutely required for RSVL's inhibition of the ability of human colon adenocarcinoma HT-29 cells to form colonies in soft agar. Binding of COX-2 with RSVL was compared with two RSVL analogues, 3,3',4',5',5-pentahydroxy-trans stilbene (RSVL-2) or 3,4',5-trimethoxy-trans-stilbene (RSVL-3). The results indicated that COX-2 binds with RSVL-2 more strongly than with RSVL, but does not bind with RSVL-3. RSVL or RSVL-2, but not RSVL-3, inhibited COX-2-mediated PGE(2) production in vitro and ex vivo. HT-29 human colon adenocarcinoma cells express high levels of COX-2 and either RSVL or RSVL-2, but not RSVL-3, suppressed anchorage independent growth of these cells in soft agar. RSVL or RSVL-2 (not RSVL-3) suppressed growth of COX-2(+/+) cells by 60% or 80%, respectively. Notably, cells deficient in COX-2 were unresponsive to RSVL or RSVL-2. These data suggest that the anticancer effects of RSVL or RSLV-2 might be mediated directly through COX-2. PMID- 18381590 TI - Coexpression of the mu-opioid receptor splice variant MOR1C and the vesicular glutamate transporter 2 (VGLUT2) in rat central nervous system. AB - It has been reported that mu-opioid agonists depress glutamate release in some neurons but the specific receptor subtype mediating this effect is unclear. The purpose of the present study was to examine whether a particular mu-opioid receptor (MOR) splice-variant, MOR(1C), is expressed in rat central nervous system (CNS) by terminals expressing the vesicular glutamate transporter2 (VGLUT2), a marker of glutamatergic neurons. Several MOR splice variants have been identified in mice and MOR(1C) appears mainly to be localized to fibers and terminals, from which most neurotransmitter release would be expected. In addition, VGLUT2 has been found in the CNS and antibodies to it are reliable markers for glutamatergic terminals. Using fluorescence immunohistochemistry and confocal microscopy to examine spatial relationships between MOR(1C) and VGLUT2, we found that MOR(1C) and VGLUT2 puncta were widely distributed throughout the rat CNS; moreover, many regions contained terminals that expressed both. Thus, it appears that coexpression of MOR(1C) and VGLUT2 is common in the rat CNS. We hypothesize that activation of MOR(1C) by mu-opioid agonists at some glutamatergic terminals may be a mechanism by which glutamate release is inhibited. PMID- 18381592 TI - Pleiotrophin inhibits transforming growth factor beta1-induced apoptosis in hepatoma cell lines. AB - Pleiotrophin (PTN) is a hepatocyte growth factor and considered to play roles in liver fibrogenesis and hepatocarcinogenesis. In this study we examined the mechanism of the action of PTN in these pathological processes. First, we confirmed that hepatic stellate cells (HSCs) and Kupffer cells, and also later hepatocytes in hyperplastic nodules increased PTN mRNA expressions during carbon tetrachloride-induced liver fibrosis. Then, the relationship between PTN and transforming growth factor beta1 (TGFbeta1), a known potent pro-fibrogenetic cytokine, in carcinogenesis was investigated using hepatoma cell lines. Huh-7 human hepatoma cells weakly expressed PTN, but HepG2 human hepatoma cells and FaO rat hepatoma cells did not. Recombinant (r) TGFbeta1 induced the cultured Huh-7 cells to undergo apoptosis, which was inhibited by rPTN. Huh-7 cells became resistant to TGFbeta1-, but not mitomycin C-induced apoptosis when transfected with PTN gene, indicating the specificity of the PTN anti-apoptotic activity. Poly ADP ribose polymerase, procaspase-8 and procaspase-3 were not cleaved in the TGFbeta1-reluctant cells. The TGFbeta1-induced caspase-3 activation was also suppressed in Huh-7 and FaO cells both transduced with PTN gene-bearing adenoviruses. In summary, PTN was expressed in HSCs, Kupffer cells, and hepatocytes in fibrotic liver. We propose that PTN specifically antagonizes the TGFbeta1 activity during liver fibrosis. PMID- 18381593 TI - SAR and efficiency evaluation of a 900 MHz waveguide chamber for cell exposure. AB - In this work we present the results of numerical and experimental dosimetry carried out for an in vitro exposure device to irradiate sample groups at 900 MHz. The cells are kept in 8 and 15 ml cell cultures, contained, respectively in T25 and T75 rectangular flasks. The dosimetric assessment of the distribution of the specific absorption rate (SAR) is performed for both the bottom of the flask and the whole volume of the sample to provide results for experiments on either the cell layer or the cell suspension. The irradiating chamber is a rectangular waveguide (WG). Different configurations are considered to assess the optimum orientation and positioning of the cell cultures inside the WG. The system performance is optimal when the electric field is parallel to the sample and the WG is terminated by a matched load. In this condition two 15 or four 8 ml cells cultures can be exposed. The efficiency (ratio between the power absorbed by the sample and the incident power) and the non-uniformity degree (ratio between the standard deviation of SAR values and the average SAR over the sample) are calculated and successfully verified through measurements of the scattering parameters and local temperature increases. In the chosen exposure configuration, the efficiency is 0.40 and the non-uniformity degree is 39% for the 15 ml samples. For the 8 ml samples, the efficiency is 0.19 and a low non-uniformity degree (15%) is found. PMID- 18381591 TI - Genome wide transcriptional profiling in breast cancer cells reveals distinct changes in hormone receptor target genes and chromatin modifying enzymes after proteasome inhibition. AB - Steroid hormone receptors, like glucocorticoid (GR) and estrogen receptors (ER), are master regulators of genes that control many biological processes implicated in health and disease. Gene expression is dependent on receptor levels which are tightly regulated by the ubiquitin-proteasome system. Previous studies have shown that proteasome inhibition increases GR, but decreases ER-mediated gene expression. At the gene expression level this divergent role of the proteasome in receptor-dependent transcriptional regulation is not well understood. We have used a genomic approach to examine the impact of proteasome activity on GR- and ER-mediated gene expression in MCF-7 breast cancer cells treated with dexamethasone (DEX) or 17beta-estradiol (E2), the proteasome inhibitor MG132 (MG) or MG132 and either hormone (MD or ME2) for 24 h. Transcript profiling reveals that inhibiting proteasome activity modulates gene expression by GR and ER in a similar manner in that several GR and ER target genes are upregulated and downregulated after proteasome inhibition. In addition, proteasome inhibition modulates receptor-dependent genes involved in the etiology of a number of human pathological states, including multiple myeloma, leukemia, breast/prostate cancer, HIV/AIDS, and neurodegenerative disorders. Importantly, our analysis reveals that a number of transcripts encoding histone and DNA modifying enzymes, prominently histone/DNA methyltransferases and demethylases, are altered after proteasome inhibition. As proteasome inhibitors are currently in clinical trials as therapy for multiple myeloma, HIV/AIDS and leukemia, the possibility that some of the target molecules are hormone regulated and chromatin modifying enzymes is intriguing in this era of epigenetic therapy. PMID- 18381594 TI - The mechanical transduction of physiological strength electric fields. AB - In this article it is proposed that electric fields of physiological strength (approximately 100 V/m) are transduced by the mechanical torque they exert on glycoproteins. The resulting mechanical signal is then transmitted to the cytoskeleton and propagated throughout the cell interior. This mechanical coupling is analyzed for transmembrane glycoproteins, such as integrins and the glycocalyx, and for glycoproteins in the extracellular matrix of cartilage. The applied torque is opposed by viscous fluid drag and restoring forces exerted by adjacent molecules in the membrane or cartilage. The resulting system represents a damped, driven harmonic oscillator. The amplitude of oscillation is constant at low frequencies, but falls off rapidly in the range 1-1000 Hz. The transition frequency depends on parameters such as the viscosity of the surrounding fluid and the restoring force exerted by the surrounding structure. The amplitude increases as the fourth power of the length of the transmembrane glycoproteins and as the square of the applied field. This process may operate in concert with other transduction mechanisms, such as the opening of voltage-gated channels and electrodiffusion/osmosis for DC fields. PMID- 18381595 TI - Exploring changes in treatment effects across design stages in adaptive trials. AB - The recently published Committee for Medicinal Products for Human Use reflection paper on flexible designs highlights a controversial issue regarding the interpretation of adaptive trials. The guideline suggests that a test for heterogeneity should be preplanned and if treatment effect estimates differ significantly between design stages then data collected before and after the interim analysis might not be combined in a formal analysis. In this paper we investigate error rates for such a procedure in the presence of calendar-time effects. Furthermore, we present an alternative testing strategy based on change point methods. In a simulation study we demonstrate that our procedure performs well in comparison to that suggested by the guideline. PMID- 18381596 TI - Osteoporosis in a Chinese population due to occupational exposure to lead. AB - BACKGROUND: Recent studies indicate that lead may exert actions both directly on osteoblast and osteoclast function, and indirectly via kidney dysfunction on bone turnover. The main focus of this study was to investigate whether occupational lead exposure is associated with low bone mass in a population working in a storage battery plant. METHODS: Monophoton absorptiometry was used to measure bone mineral density (BMD) in the population and the Z score was introduced to define osteoporosis (Z score <-2). Lead concentration of urine and blood was determined by graphite-furnace atomic absorption spectrophotometry as an exposure biomarker. A total of 249 persons (191 males and 58 females) participated and completed a questionnaire in order to obtain information on height, weight, age, medical and drug history, cigarette smoking, alcohol consumption, job position, work year, physical exercise, etc. RESULTS: The BMD was significantly decreased in the groups of the high urinary lead (UPb) level compared with the low UPb level with a significant difference (P < 0.05) in both genders, but no such significant difference was observed in the relationship between blood lead (BPb) and BMD. The prevalence of osteoporosis would increase significantly with the increase of the UPb (P < 0.01) in the linear correlation manner (P < 0.01). There was also such a significant relationship between BPb and osteoporosis (P < 0.01). There was a dose-response relationship between lead exposure and prevalence of osteoporosis. CONCLUSIONS: In contrast to BPb, UPb had a more close relationship with osteoporosis caused by lead. It was concluded that occupational exposure to lead is associated with osteoporosis. PMID- 18381597 TI - Disparities in work-related injuries associated with worker compensation coverage status. AB - BACKGROUND: This exploratory study addresses patterns of injury in an emerging population of contingent workers who are not covered by either worker's compensation (WC) or health insurance. The primary purpose is to improve the information base regarding the entire population of uninsured, injured workers. Because Latino workers are over-represented in the uninsured group, we include additional characterization of their patterns of injury. Recent studies have found that worker compensation claims and reports address a shrinking proportion of occupational injury and exposure, and about two-thirds of occupational injuries are not captured in the U.S. national surveillance system. METHODS: Following the NEISS methodology, a work-relatedness indicator was retrieved for emergency department (ED) visits to an academic health center in fiscal year 2005. RESULTS: Twenty percent of self-declared work-related injuries were not associated with self-reported WC coverage. Parametric and non-parametric statistical analysis found several significant disparities in workers without WC. These disparities included a higher proportion of Latinos, workers under age 25, and construction workers. In the uninsured group, Latino workers had a higher proportion of moderate and severe injuries. Nearly all (92 percent) workers without WC also lacked health insurance. Injured low-income workers who lack access to both WC and employer-sponsored health insurance comprise an increasing percentage of the occupationally injured. Our exploratory study found this to be particularly true in high-risk populations. CONCLUSIONS: Work-relatedness indicators collected routinely in ED and outpatient settings should be incorporated into standard reporting systems to facilitate more accurate and comprehensive surveillance and better-targeted interventions. PMID- 18381598 TI - Statisticians and evidence--mote and beam. PMID- 18381599 TI - Visual, lateral line, and auditory ascending pathways to the dorsal telencephalic area through the rostrolateral region of the lateral preglomerular nucleus in cyprinids. AB - Fiber connections of the rostrolateral region of the lateral preglomerular nucleus (PGlr) were studied by tract-tracing methods in carp and goldfish. The PGlr receives fibers from the optic tectum, ventrolateral nucleus of semicircular torus, ventromedial thalamic nucleus, medial pretoral nucleus, anterior tuberal nucleus, subglomerular nucleus, and (unexpectedly) also from the retina. Dendritic morphology of tecto-preglomerular neurons suggests that they receive retinal inputs. The PGlr can be further subdivided into dorsal (PGlr-d) and ventral (PGlr-v) zones, both of which are composed of somata and neuropil layers. Retinal and tectal fibers terminate mostly in the neuropil layer of the PGlr-d with the retinal terminals concentrated medially and tectal terminals laterally. Lateral line toral fibers terminate mainly in a lateral portion and ventromedial thalamic fibers in a medial portion of the somata layer of the PGlr-d. Auditory fibers from the medial pretoral nucleus and anterior tuberal nucleus terminate in the PGlr-v. The central nucleus of the semicircular torus also projects sparse fibers to the PGlr-v. The PGlr projects to the lateral, central, and medial parts of the dorsal telencephalic area, and the latter telencephalic part sends descending fibers to the PGlr. Differential distribution patterns of PGlr-d and PGlr-v fibers are noted within the dorsal telencephalic parts, suggesting that different sensory modalities may be represented in distinct regions at least to a certain degree. PMID- 18381600 TI - Extremely low frequency (ELF) electric and magnetic field exposure limits: rationale for basic restrictions used in the development of an Australian standard. AB - There are large disparities between basic restrictions for exposure to extremely low-frequency (0-3 kHz) Electric and Magnetic Fields set by two major international bodies. Both bodies agree that these basic restrictions should prevent neuro-stimulatory effects: the retinal phosphene at frequencies up to a few hundred Hertz and peripheral nervous stimulation (PNS) at higher frequencies. The disparity arises from differences in estimated thresholds and frequency dependence, and whether restrictions should be of tissue induced current density or electric field. This paper argues that the latter metric more directly relates to neurostimulatory processes. By analysing available literature, a threshold for retinal phosphenes occurrence is found to be 56 mV/m (95% Confidence Interval 2 1330 mV/m), with a characteristic frequency of 20 Hz. Similarly, the smallest PNS sensation threshold is identified at 2 V/m (characteristic frequency above 3 kHz). In the case of the former, the large range of uncertainty suggests a 'power of ten' value of 100 mV/m. For the latter, because of the small margin between sensation and pain threshold, and because of the large individual variation, the smallest estimate of sensation threshold (2 V/m) represents a basic restriction with precaution incorporated. PMID- 18381603 TI - Long-term survival after radical prostatectomy compared to other treatments in older men with local/regional prostate cancer. AB - BACKGROUND: This study aimed to address long-term survival in a large population based cohort of men with prostate cancer receiving radical prostatectomy compared to other treatments. METHODS: We studied 5,845 patients diagnosed with local/regional stage prostate cancer at age 65-74 in 1992 with comorbidity score <2, who were defined as potential candidates for radical prostatectomy and identified from the SEER (Surveillance, Epidemiology and End Results)-Medicare cohort with median follow-up of 11 years. RESULTS: Of 5,845 patients, 10-year all cause survival rates were the highest for patients receiving radical prostatectomy (81.0%; 95% CI: 79.4-82.4%), followed by radical prostatectomy in combination with radiotherapy (67.6%; 62.0-72.5%), radiotherapy (60.5%; 58.3 62.6%), and were the lowest for watchful-waiting (50.7%; 47.5-53.8%). A similar pattern was found for 10-year prostate cancer-specific survivals by treatments. After adjusting for age, ethnicity, region, Gleason Score, comorbidity, median annual household income, hormone therapy and chemotherapy, the hazard ratio of all-cause mortality was 0.31 (95% CI: 0.25-0.37) for radical prostatectomy and 0.38 (95% CI: 0.28-0.52) for radical prostatectomy plus radiation therapy compared to those with watchful-waiting. CONCLUSIONS: There was a significant long-term survival benefit in men receiving radical prostatectomy compared to those receiving watchful-waiting or radiotherapy. PMID- 18381601 TI - Inhibition of SNAP25 expression by HIV-1 Tat involves the activity of mir-128a. AB - MicroRNAs (miRs) are short endogenous RNAs that regulate gene expression by incomplete pairing with messenger RNAs. An increasing number of studies show that mammalian microRNAs play fundamental roles in various aspects of cellular function including differentiation, proliferation, and cell death. Recent findings demonstrating the presence of microRNAs in mature neuronal dendrites suggest their possible involvement in controlling local protein translation and synaptic function. HIV-1 Encephalopathy (HIVE) is a manifestation of HIV-1 infection that often results in neuronal damage and dysfunction. While neurons are rarely, if ever, infected by HIV-1, they are exposed to cytotoxic viral and cellular factors including the HIV-1 transactivating factor Tat. In this study, we show that Tat deregulates expression levels of selected microRNAs, including the neuronal mir-128, in primary cortical neurons. We further show that mir-128a inhibits expression of the pre-synaptic protein SNAP25, whereas the anti-mir-128a partially restores Tat/mir-128a-induced downregulation of SNAP25 expression. Altogether, our data provide a novel mechanism by which HIV-Tat perturbs neuronal activity. PMID- 18381602 TI - Synergistic anti-proliferative and pro-apoptotic activity of combined therapy with bortezomib, a proteasome inhibitor, with anti-epidermal growth factor receptor (EGFR) drugs in human cancer cells. AB - The proteasome plays a pivotal role in the turnover of regulatory transduction proteins induced by activated cell membrane growth factor receptors. The epidermal growth factor receptor (EGFR) pathway is crucial in the development and progression of human epithelial cancers. Proteasome inhibition may sensitize human cancer cell lines to EGFR inhibitors. We investigated the growth inhibitory and pro-apoptotic effects of the proteasome inhibitor bortezomib in combination with anti-EGFR drugs, such as gefitinib, vandetanib, and cetuximab in EGFR expressing human cancer cell lines. Bortezomib determined dose-dependent growth inhibition in a nine cancer cell line panel (IC(50) values, range 6-42 nM). A significant synergistic growth inhibitory effect was observed with the combination of bortezomib and each EGFR inhibitor in all cell lines (combination index, CI, range 0.10-0.55), which was accompanied by a significant induction in apoptosis by the combined treatment with bortezomib, cetuximab and vandetanib. In HCT-116 colon cancer and A549 lung adenocarcinoma cells, bortezomib plus EGFR inhibitor treatment induced a more effective inhibition of EGFR-activated down stream signals, including a marked suppression in activated, phosphorylated Akt (P-Akt). In contrast, overexpression of a constitutively active P-Akt protected A549 cells by cell growth inhibition and apoptosis following treatment with bortezomib and EGFR inhibitors. The combined treatment with bortezomib and EGFR inhibitors has a synergistic growth inhibitory and pro-apoptotic activity in different human cancer cells which possess a functional EGFR-dependent autocrine growth pathway through to a more efficient and sustained inhibition of Akt. PMID- 18381604 TI - Genetic variants in germline TP53 and MDM2 SNP309 are not associated with early onset colorectal cancer. AB - BACKGROUND AND OBJECTIVES: Colorectal cancer (CRC) arising in patients under age 30 is a rare disease, and few cases have been reported within Li-Fraumeni kindreds. To determine how often alterations in the p53 pathway genes contribute to disease susceptibility, we have evaluated patients with early onset CRC for the presence of germline variants in the p53 gene and MDM2 SNP309. METHODS: Thirty-five patients with CRC diagnosed before age 30 were included in this study based on tissue availability. DNA samples from peripheral blood leukocytes were analyzed for constitutional mutations and polymorphisms in p53 as well as polymorphisms in MDM2 SNP309. RESULTS: No mutations were found in exons 4-10 of the p53 gene. The frequencies of polymorphisms in p53 and in MDM2 SNP309 did not differ from rates previously reported for normal control populations, and no polymorphism in either gene could be associated with early onset CRC. CONCLUSIONS: Neither germline variants in p53 nor MDM2 SNP309 play an underlying role in the development of very early onset CRC. For the large majority of cases, the genetic basis of this disease remains unknown. PMID- 18381605 TI - Occurrence and cellular localization of PrPd in kidneys of scrapie-affected sheep in the absence of inflammation. AB - Following a preliminary description of disease-associated prion protein (PrPd) deposition in the kidneys of scrapie-affected sheep, detailed studies have been undertaken in order to evaluate the factors that could account for such PrPd accumulation and to determine the precise location of PrPd in the renal papillae. Immunohistochemical (IHC) examinations for PrPd were conducted in kidneys collected at post-mortem from 30 naturally and 37 experimentally infected sheep. In addition, PrPd detection by western blot analysis (WB) and ultrastructural examination was carried out in a selection of kidneys. PrPd-specific, multifocal IHC labelling with antibody R145 was achieved in the kidneys of 44% and 51% of the naturally and experimentally infected sheep, respectively. The specificity of these results was confirmed by further IHC and WB using several PrP antibodies raised to different amino acid sequences, and by examination of control tissues. PrPd was shown to accumulate in the interstitium of the renal papillae, in association with the cell membrane and lysosomes of fibroblast-like cells, or extracellularly, in close contact with collagen and basal membranes. These deposits were unrelated to inflammatory changes in the kidney as shown by routine histology and by IHC for different immune cell markers. PrPd accumulated in the kidney of sheep that showed widespread PrPd deposition in the lymphoreticular system and had long incubation periods; these findings argue for a haematogenous origin of renal PrPd, although the precise site and mechanism-glomerular filtration and reabsorption at Henle's loop, or extravasation from vasa recta capillaries, or both-by which PrPd leaves the blood to accumulate in the interstitium of renal papillae remain to be determined. Either of these pathogenetic mechanisms could lead to environmental contamination via urine. PMID- 18381606 TI - Catalytic C-C, C-N, and C-O Ullmann-type coupling reactions: copper makes a difference. PMID- 18381607 TI - Intermolecular forces in an ionic liquid ([Mmim][Cl]) versus those in a typical salt (NaCl). PMID- 18381608 TI - Non-aqueous routes to metal oxide thin films by atomic layer deposition. PMID- 18381609 TI - Fast cis-trans isomerization of an azobenzene derivative in liquids and liquid crystals under a low electric field. PMID- 18381610 TI - Cardiac liposarcoma and bilateral ovarian mature teratoma in a patient with congenital heart disease and f-anisosplenia. AB - A 10-year-old female with atrial septal defect presented with bilateral ovarian mature cystic teratomas. Three spleniculi adjacent to the main spleen were noticed while the endoscopic surgery. Two years later a heart tumor was discovered which led to death. Pathologic examination of samples of the mass proved it to be a liposarcoma. The unusual combination of atrial septal defect, f anisosplenia (female, congenital heart disease, multiple unevenly-sized spleens, left lung type pulmonary isomerism), bilateral ovarian mature cystic teratomas and liposarcoma of the heart appears to be a unique presentation. PMID- 18381611 TI - Investigation of associations between the pregnane-X receptor gene (NR1I2) and Crohn's disease in Canadian children using a gene-wide haplotype-based approach. AB - BACKGROUND: The pregnane-X-receptor (PXR) is involved in the metabolism and detoxification of numerous xenobiotics and is critical for maintaining intestinal integrity. The NR1I2 gene encoding PXR may confer susceptibility for Crohn's disease (CD) but evidence for associations is conflicting. We investigated whether the NR1I2 gene was associated with susceptibility for pediatric CD. METHODS: A case-control and family-based (case-parent) study was carried out at 3 inflammatory bowel disease (IBD) clinics across Canada. Confirmed cases of CD <20 years were diagnosed using standard criteria. For determination of gene associations parents of the cases and unrelated controls were evaluated. Clinical phenotypes were established based on the Montreal Classification scheme. Eight tag-SNPs (tag-single nucleotide polymorphisms) across the gene were genotyped for allelic or genotypic associations. RESULTS: A total of 270 CD cases, 336 controls, and 395 parents were studied. The mean age (+/-SD) of the cases was 12.1 (+/-3.5) years of age. Most cases were male (56.3%), had disease location L3+/-L4 (58.1%), and an inflammatory phenotype B1+/-p (88.4%) at diagnosis. For 7 SNPs single SNP analysis using case-control or case-parent data did not reveal associations with development of CD and none of the SNPs were significantly associated with disease location or disease behavior at diagnosis. One SNP rs2461823 (P = 0.05) was nominally associated with CD. No overall haplotype association (omnibus P-value = 0.61) or associations with individual haplotypes was evident. CONCLUSIONS: Our gene-wide analysis in a pediatric cohort using both the case-control and case-parent designs suggests that the NR1I2 gene is not associated with CD in Canadian children. PMID- 18381612 TI - Ileouterine fistula: an unusual complication of Crohn's disease. PMID- 18381613 TI - A multicenter study on the prevalence and spectrum of mutations in the otoferlin gene (OTOF) in subjects with nonsyndromic hearing impairment and auditory neuropathy. AB - Autosomal recessive nonsyndromic hearing impairment (NSHI) is a heterogeneous condition, for which 53 genetic loci have been reported, and 29 genes have been identified to date. One of these, OTOF, encodes otoferlin, a membrane-anchored calcium-binding protein that plays a role in the exocytosis of synaptic vesicles at the auditory inner hair cell ribbon synapse. We have investigated the prevalence and spectrum of deafness-causing mutations in the OTOF gene. Cohorts of 708 Spanish, 83 Colombian, and 30 Argentinean unrelated subjects with autosomal recessive NSHI were screened for the common p.Gln829X mutation. In compound heterozygotes, the second mutant allele was identified by DNA sequencing. In total, 23 Spanish, two Colombian and two Argentinean subjects were shown to carry two mutant alleles of OTOF. Of these, one Colombian and 13 Spanish subjects presented with auditory neuropathy. In addition, a cohort of 20 unrelated subjects with a diagnosis of auditory neuropathy, from several countries, was screened for mutations in OTOF by DNA sequencing. A total of 11 of these subjects were shown to carry two mutant alleles of OTOF. In total, 18 pathogenic and four neutral novel alleles of the OTOF gene were identified. Haplotype analysis for markers close to OTOF suggests a common founder for the novel c.2905_2923delinsCTCCGAGCGCA mutation, frequently found in Argentina. Our results confirm that mutation of the OTOF gene correlates with a phenotype of prelingual, profound NSHI, and indicate that OTOF mutations are a major cause of inherited auditory neuropathy. PMID- 18381615 TI - Determination of selected non-authorized insecticides in peppers by liquid chromatography time-of-flight mass spectrometry and tandem mass spectrometry. AB - In this work, two analytical methods based on liquid chromatography coupled to electrospray time-of-flight mass spectrometry (LC/ESI-TOFMS) and tandem mass spectrometry (LC/ESI-MS/MS) are described for the identification, confirmation and quantitation of three insecticides non-authorized in the European Union (nitenpyram, isocarbophos and isofenphos-methyl) but detected in recent monitoring programmes in pepper samples. The proposed methodologies involved a sample extraction procedure using liquid-liquid partition with acetonitrile followed by a cleanup step based on dispersive solid-phase extraction. Recovery studies performed on peppers spiked at different fortification levels (10 and 50 microg kg(-1)) yielded average recoveries in the range 76-100% with relative standard deviation (RSD) (%) values below 10%. Identification, confirmation and quantitation were carried out by LC/TOFMS and LC/MS/MS using a hybrid triple quadrupole linear ion trap (QqLIT) instrument in multiple-reaction monitoring (MRM) mode. The obtained limits of quantitation (LOQs) were in the range 0.1-5 microg kg(-1), depending on each individual technique. Finally, the proposed methods were successfully applied to the analysis of suspected pepper samples. PMID- 18381614 TI - Bacteria and bacterial rRNA genes associated with the development of colitis in IL-10(-/-) mice. AB - BACKGROUND: Microorganisms appear to play important yet ill-defined roles in the etiology of inflammatory bowel disease (IBD). This study utilized a novel population-based approach to identify bacteria and bacterial rRNA genes associated with the development of colitis in IL-10(-/-) mice. METHODS: Mice were housed in 2 environments: a community mouse facility where the mice were fed nonsterile chow (Room 3) and a limited access facility where the mice were fed sterile chow (Room 4). Every month the disease activity levels were assessed and fecal bacterial compositions were analyzed. At the end of the experiments histological and bacterial analyses were performed on intestinal tissue. RESULTS: Although disease activity increased over time in both environments, it progressed at a faster rate in Room 3 than Room 4. Culture and culture-independent bacterial analyses identified several isolates and phylotypes associated with colitis. Two phylotypes (GpC2 and Gp66) were distinguished by their negative associations with disease activity in fecal and tissue samples. Notably, rRNA genes from these phylotypes had high sequence identity (99%) to an rRNA gene from a previously described flagellated Clostridium (Lachnospiraceae bacterium A4). CONCLUSIONS: The negative associations of these 2 phylotypes (GpC2 and Gp66) suggest that these bacteria were being immunologically targeted, consistent with prior findings that the Lachnospiraceae bacterium A4 bears a prevalent flagellar antigen for disease-associated immunity in murine immune colitis and human Crohn's disease. Identification of these associations suggests that the experimental approach used in this study will have considerable utility in elucidating the host-microbe interactions underlying IBD. PMID- 18381616 TI - In vivo MR imaging of magnetically labeled mesenchymal stem cells transplanted into rat liver through hepatic arterial injection. AB - OBJECTIVE: The aim of this study was to evaluate in vivo magnetic resonance imaging (MRI) for tracking the magnetically labeled mesenchymal stem cells (MSCs) transplanted into rat liver through hepatic arterial injection. MATERIALS AND METHODS: MSCs, harvested from the bone marrow of Wistar rats and expanded by the adhesion method, were labeled with both Feridex and 4',6-diamidino-2-phenylindole (DAPI). Cell transplantation was performed by injection of 1 x 10(6) labeled cells (n = 20) or unlabeled cells (n = 10) via hepatic artery into rat livers treated with 2% carbon tetrachloride to induce acute liver necrosis. MR imaging was performed on a clinical 1.5 T MR scanner with a T(2)*-weighted gradient-echo sequence immediately before and at 1 h, 3 days, 7 days and 14 days after transplantation, and the signal-to-noise ratios (SNRs) were measured in liver, spleen, kidney and muscle. After MR examination, the animals were sacrificed, and the liver, kidney, lung and muscle were prepared for fluorescence observation and Prussian Blue staining. RESULTS: In the group treated with labeled cells, the SNR of the liver after cell transplantation was 3.12 +/- 0.43 at 1 h, 7.98 +/- 1.05 at 3 days and 11.46 +/- 1.41 at 7 days. These values were significantly lower than the pre-transplantation SNR (14.40 +/- 0.37). In the group treated with unlabeled cells, no significant difference could be found between after and before transplantation liver SNRs. Prussian Blue staining showed iron particles, contained within the cytoplasm and distributed in the liver parenchyma, which corresponded to the DAPI-stained fluorescent nuclei under the fluorescence microscope. CONCLUSION: The magnetically labeled MSCs transplanted into rat liver through hepatic arterial injection can be detected and monitored in vivo with a 1.5 T clinical MR scanner for up to 7 days after cell transplantation. PMID- 18381617 TI - Functional and phenotypic characteristics of testicular macrophages in experimental autoimmune orchitis. AB - Testicular inflammation with compromised fertility can occur despite the fact that the testis is considered an immunoprivileged organ. Testicular macrophages have been described as cells with an immunosuppressor profile, thus contributing to the immunoprivilege of the testis. Experimental autoimmune orchitis (EAO) is a model of organ-specific autoimmunity and testicular inflammation. EAO is characterized by an interstitial inflammatory mononuclear cell infiltration, damage of the seminiferous tubules and germ cell apoptosis. Here we studied the phenotype and functions of testicular macrophages during the development of EAO. By stereological analysis, we detected an increased number of resident (ED2+) and non-resident (ED1+) macrophages in the testicular interstitium of rats with orchitis. We showed that this increase was mainly due to monocyte recruitment. The in vivo administration of liposomes containing clodronate in rats undergoing EAO led to a reduction in the number of testicular macrophages, which correlated with a decreased incidence and severity of the testicular damage and suggests a pathogenic role of macrophages in EAO. By immunohistochemistry and flow cytometry we detected an increased number of testicular macrophages expressing MHC class II, CD80 and CD86 costimulatory molecules in rats with orchitis. Also, testicular macrophages from rats with EAO showed a higher production of IFNgamma (ELISA). We conclude that testicular macrophages participate in EAO development, and the ED1+ macrophage subset is the main pathogenic subpopulation. They stimulate the immune response through the production of pro-inflammatory cytokines and antigen presentation and thus activation of T cells in the target organ. PMID- 18381618 TI - Non-labeled detection of waterborne pathogen Cryptosporidium parvum using a polydiacetylene-based fluorescence chip. AB - A non-labeling fluorescence sensor system was developed using polydiacetylene (PDA) liposomes composed of 10,12-pentacosadiynoic acid (PCDA) and 1,2 dimyristoyl-sn-glycero-3-phosphocholine (DMPC) at a 8:2 molar ratio. The PDA liposomes were immobilized onto an amine-coated glass surface using peptide bonding between the carboxyl group of the liposome and the amine group of the glass surface. The optimum ratio of the cross linker (NHS/EDC) to PDA liposome was determined to be 50% for strong immobilization of the liposomes. Residual carboxyl groups of the PDA liposomes were selectively biotinylated, followed by sequential binding of streptavidin and biotin-antibody (bioreceptor). Finally, the performance of the PDA liposome chip was tested for detecting Cryptosporidium parvum, and yielded a detection limit of 1 x 10(3) oocysts/mL. From these results, it is expected that the PDA liposome chip will have high application potential for the detection of waterborne pathogens including C. parvum. PMID- 18381619 TI - Effects of electrospray capillary temperature on amide hydrogen exchange. AB - Amide hydrogen/deuterium (H/D) exchange coupled with proteolysis, high perfeomance liquid chromatographic (HPLC) separation and mass spectrometry (MS) has become a powerful tool to study protein dynamics in solution. Prior to the execution of H/D exchange experiments, various experimental parameters have to be set, including proteolysis, HPLC, and MS conditions. Here we investigate the effects of electrospray capillary temperature on deuterium retention in backbone amides of various pepsin-generated cytochrome c peptides. Lower capillary temperature generally helps retain more deuterium than higher capillary temperature. When the capillary temperature was 150 degrees C, on average 26% more deuterium was retained than when the capillary temperature was set at 250 degrees C. The effects of capillary temperature varied depending on the ions monitored. There was little difference in deuterium retention among different charge state species of the same peptide at 150 degrees C. However, a lower charge state ion loses more deuterium atoms going from 150 degrees C to 250 degrees C than the corresponding higher charge state species. These results indicate that the capillary temperature should be optimized not only to maximize the signal-to-noise of each ion followed in H/D exchange experiments, but also to minimize the deuterium loss of the ions. Also the loss of deuterium in several ions, especially lower charge state ones, should be monitored in the optimization, as the temperature effects vary among ions and are more significant for lower charge state ions. PMID- 18381620 TI - Application of ion trap technology to liquid chromatography/mass spectrometry quantitation of large peptides. AB - Triple quadrupole mass spectrometers are generally considered the instrument of choice for quantitative analysis. However, for the analysis of large peptides we have encountered some cases where, as the data presented here would indicate, ion trap mass spectrometers may be a good alternative. In general, specificity and sensitivity in bioanalytical liquid chromatography/mass spectrometry (LC/MS) assays are achieved via tandem MS (MS/MS) utilizing collision-induced dissociation (CID) while monitoring unique precursor to product ion transitions (i.e. selected reaction monitoring, SRM). Due to the difference in CID processes, triple quadrupoles and ion traps often generate significantly different fragmentation spectra of product ion species and intensities. The large peptidic analytes investigated here generated fewer fragments with higher relative abundance on the ion trap as compared to those generated on the triple quadrupole, resulting in lower limits of detection on the ion trap. PMID- 18381621 TI - Sub one minute inhibition assays for the major cytochrome P450 enzymes utilizing ultra-performance liquid chromatography/tandem mass spectrometry. AB - The measurement of cytochrome P450 (CYP450) isoenzyme inhibition is often done during evaluation of new chemical entities in drug discovery. Typical assay protocol consists of multiple CYP450 probe substrates incubated with selected drug candidates and CYP450. Results of the assay, the amount of probe substrate metabolite formed with respect to control, are used to determine the level of interaction. Liquid chromatography utilizing columns packed with sub-2-micron particles have been shown to provide up to 8X faster analysis time and 3X increases in sensitivity over traditional high-performance liquid chromatography (HPLC). The work presented here shows the development of a high-throughput, sub-2 micron particle LC method coupled with tandem quadrupole mass spectrometry for the rapid analysis of six CYP450 probe substrate metabolites in 30s. PMID- 18381622 TI - Novel possibilities in the study of isolated carbon nanotubes. AB - We report the first detection of carbon nanotubes in the gas phase in a matrix assisted laser desorption/ionization (MALDI) mass spectrometric experiment. These observations open the possibility of studying isolated nanotubes in the gas phase by means of various spectroscopic methods and a possible way to separate them. PMID- 18381623 TI - A multi-angular mass spectrometric view at cyclic nucleotide dependent protein kinases: in vivo characterization and structure/function relationships. AB - Mass spectrometry has evolved in recent years to a well-accepted and increasingly important complementary technique in molecular and structural biology. Here we review the many contributions mass spectrometry based studies have made in recent years in our understanding of the important cyclic nucleotide activated protein kinase A (PKA) and protein kinase G (PKG). We both describe the characterization of kinase isozymes, substrate phosphorylation, binding partners and post translational modifications by proteomics based methodologies as well as their structural and functional properties as revealed by native mass spectrometry, H/D exchange MS and ion mobility. Combining all these mass spectrometry based data with other biophysical and biochemical data has been of great help to unravel the intricate regulation of kinase function in the cell in all its magnificent complexity. PMID- 18381624 TI - Cell penetrating peptide-modified pharmaceutical nanocarriers for intracellular drug and gene delivery. AB - Cell-penetrating peptides (CPPs) including TAT peptide (TATp) have been successfully used for intracellular delivery of a broad variety of cargos including various nanoparticulate pharmaceutical carriers (liposomes, micelles, nanoparticles). Here, we will consider the main results in this area, with a special emphasis on TATp-mediated delivery of liposomes and DNA. We will also address the development of "smart" stimuli-sensitive nanocarriers, where cell penetrating function can be activated by the decreased pH only inside the biological target minimizing thus the interaction of drug-loaded nanocarriers with nontarget cells. PMID- 18381625 TI - Synthesis and in vivo studies of carbohydrate-based vaccines against group A streptococcus. AB - Carbohydrates, as carriers, providing numerous attachment points for the conjugation of peptide antigens and their optimal orientation for the recognition by cells of the immune system, reducing degradation of the attached peptide antigens and many other advantages make carbohydrate-based vaccine highly promising approach. Multiple copies of a single group A streptococcal (GAS) M protein derived specific peptide antigens (J8 or J14) were coupled onto carbohydrate cores (D-glucose and D-galactose) linked to lipophilic amino acids to produce a self-adjuvanting liposaccharide vaccine against GAS strains. In vivo experiments showed high serum IgG antibody titers against each of the incorporated peptide epitopes, J8 or J14. PMID- 18381627 TI - Conformational studies of hexapeptides containing two dehydroamino acid residues in positions 2 and 5 in peptide chain. AB - Conformational preferences of a group of hexapeptides containing two dehydroamino acid residues in Positions 2 and 5 in peptide chain were investigated by means of spectroscopic methods (NMR and CD) and theoretical calculations. In the case of dimethylsulfoxide (DMSO) solution, only peptide with free N-termini adopted rigid 3(10)-helical conformation, for the rest of examined peptides extended and "zig zag" conformers were predominant. CD measurements showed that only in chloroform solution the conformational freedom of investigated peptides was restricted. PMID- 18381626 TI - A poke in the eye: inhibiting HIV-1 protease through its flap-recognition pocket. AB - A novel mechanism of inhibiting HIV-1 protease (HIVp) is presented. Using computational solvent mapping to identify complementary interactions and the Multiple Protein Structure method to incorporate protein flexibility, we generated a receptor-based pharmacophore model of the flexible flap region of the semiopen, apo state of HIVp. Complementary interactions were consistently observed at the base of the flap, only within a cleft with a specific structural role. In the closed, bound state of HIVp, each flap tip docks against the opposite monomer, occupying this cleft. This flap-recognition site is filled by the protein and cannot be identified using traditional approaches based on bound, closed structures. Virtual screening and dynamics simulations show how small molecules can be identified to complement this cleft. Subsequent experimental testing confirms inhibitory activity of this new class of inhibitor. This may be the first new inhibitor class for HIVp since dimerization inhibitors were introduced 17 years ago. PMID- 18381628 TI - Do crocodilians get the flu? Looking for influenza A in captive crocodilians. AB - It is well established that several wild aquatic bird species serve as reservoirs for the influenza A virus. It has also been shown that the influenza A virus can be transmitted to mammalian species such as tigers and domestic cats and dogs through ingestion of infected birds. Another group of animals that should also be considered as potential hosts for the influenza A virus are the crocodilians. Many crocodilian species share aquatic environments with wild birds that are known to harbor influenza viruses. In addition, many large crocodilians utilize birds as a significant food source. Given these factors in addition to the close taxonomic proximity of aves to the crocodilians, it is feasible to ask whether crocodilian species may also harbor the influenza A virus. Here we analyzed 37 captive crocodilians from two locations in Florida (plus 5 wild bird fecal samples from their habitat) to detect the presence of influenza A virus. Several sample types were examined. Real-time RT-PCR tests targeting the influenza A matrix gene were positive for four individual crocodilians--Alligator sinensis, Paleosuchus trigonatus, Caiman latirostris and Crocodylus niloticus. Of the seven serum samples tested with the avian influenza virus agar gel immunodiffusion assay, three showed a nonspecific reaction to the avian influenza virus antigen A. sinensis, P. trigonatus and C. niloticus (C. latirostris was not tested). Viable virus could not be recovered from RT-PCR-positive samples, although this is consistent with previous attempts at viral isolation in embryonated chicken eggs with crocodilian viruses. PMID- 18381629 TI - Sleeping tree choice by Bwindi chimpanzees. AB - Unlike nearly all other nonhuman primates, great apes build sleeping nests. In Bwindi Impenetrable National Park, Uganda, chimpanzees build nests nightly and also build day nests. We investigated patterns of nest tree use by Bwindi chimpanzees to understand ecological influences on nest tree selection. We analyzed data on 3,414 chimpanzee nests located from 2000 to 2004. Chimpanzees at Bwindi were selective in their use of nest trees. Of at least 163 tree species known to occur in Bwindi [Butynski, Ecological survey of the Impenetrable (Bwindi) Forest, Uganda, and recommendations for its conservation and management. Report to the Government of Uganda, 1984], chimpanzees utilized only 38 species for nesting. Of these, four tree species (Cassipourea sp., Chrysophyllum gorungosanum, Drypetes gerrardii, and Teclea nobilis) accounted for 72.1% of all nest trees. There was considerable variation in nesting frequencies among the top four species between and within years. However, these species were used significantly more often for nesting than other species in 70.9% (39 of 55) of the months of this study. A Spearman rank correlation found no significant relationship between tree abundance and tree species preference. Ninety-three percent of all nests were constructed in food tree species, although not necessarily at the same time the trees bore food items used by chimpanzees. The results indicate that nesting tree species preferences exist. Bwindi chimpanzees' choice of nesting tree species does not appear to be dependent on tree species density or use of the tree for food. We discuss possible reasons for the selectivity in nest trees by the Bwindi population. PMID- 18381630 TI - Using grey dynamic modeling and pseudo amino acid composition to predict protein structural classes. AB - Using the pseudo amino acid (PseAA) composition to represent the sample of a protein can incorporate a considerable amount of sequence pattern information so as to improve the prediction quality for its structural or functional classification. However, how to optimally formulate the PseAA composition is an important problem yet to be solved. In this article the grey modeling approach is introduced that is particularly efficient in coping with complicated systems such as the one consisting of many proteins with different sequence orders and lengths. On the basis of the grey model, four coefficients derived from each of the protein sequences concerned are adopted for its PseAA components. The PseAA composition thus formulated is called the "grey-PseAA" composition that can catch the essence of a protein sequence and better reflect its overall pattern. In our study we have demonstrated that introduction of the grey-PseAA composition can remarkably enhance the success rates in predicting the protein structural class. It is anticipated that the concept of grey-PseAA composition can be also used to predict many other protein attributes, such as subcellular localization, membrane protein type, enzyme functional class, GPCR type, protease type, among many others. PMID- 18381631 TI - Induction correction model for rotation of two or three dihedral angles. AB - In a previous work we have introduced an intramolecular induction correction model. In this work we have used the model to calculate the total dipol moment of six molecules as a function of two or three dihedral angles that are simultaneously varied in the molecules. It is found that the induction model behaves very well for the systems studied when compared with a regular force field model where fixed charges and dipoles are rotated along with the atoms of the molecules. This suggests that the proposed induction correction model can be used to model systems containing several dihedral angles around which rotations are performed. PMID- 18381632 TI - Ion flux through membrane channels--an enhanced algorithm for the Poisson-Nernst Planck model. AB - A novel algorithmic scheme for numerical solution of the 3D Poisson-Nernst-Planck model is proposed. The algorithmic improvements are universal and independent of the detailed physical model. They include three major steps: an adjustable gradient-based step value, an adjustable relaxation coefficient, and an optimized segmentation of the modeled space. The enhanced algorithm significantly accelerates the speed of computation and reduces the computational demands. The theoretical model was tested on a regular artificial channel and validated on a real protein channel-alpha-hemolysin, proving its efficiency. PMID- 18381635 TI - Activation of olfactory and trigeminal cortical areas following stimulation of the nasal mucosa with low concentrations of S(-)-nicotine vapor--an fMRI study on chemosensory perception. AB - Applied to the nasal mucosa in low concentrations, nicotine vapor evokes odorous sensations (mediated by the olfactory system) whereas at higher concentrations nicotine vapor additionally produces burning and stinging sensations in the nose (mediated by the trigeminal system). The objective of this study was to determine whether intranasal stimulation with suprathreshold concentrations of S(-) nicotine vapor causes brain activation in olfactory cortical areas or if trigeminal cortical areas are also activated. Individual olfactory detection thresholds for S(-)-nicotine were determined in 19 healthy occasional smokers using a computer-controlled air-dilution olfactometer. Functional magnetic resonance images were acquired using a 1.5T MR scanner with applications of nicotine in concentrations at or just above the individual's olfactory detection threshold. Subjects reliably perceived the stimuli as being odorous. Accordingly, activation of brain areas known to be involved in processing of olfactory stimuli was identified. Although most of the subjects never or only rarely observed a burning or painful sensation in the nose, brain areas associated with the processing of painful stimuli were activated in all subjects. This indicates that the olfactory and trigeminal systems are activated during perception of nicotine and it is not possible to completely separate olfactory from trigeminal effects by lowering the concentration of the applied nicotine. In conclusion, even at low concentrations that do not consistently lead to painful sensations, intranasally applied nicotine activates both the olfactory and the trigeminal system. PMID- 18381633 TI - Dynorphin and stress-related peptides in rat locus coeruleus: contribution of amygdalar efferents. AB - The interaction between the stress axis and endogenous opioid systems has gained substantial attention, because it is increasingly recognized that stress alters individual sensitivity to opiates. One site at which opiates and stress substrates may interact to have global effects on behavior is within the locus coeruleus (LC). We have previously described interactions of several opioid peptides [e.g., proopiomelanocortin, enkephalin (ENK)] with the stress-related peptide corticotropin-releasing factor (CRF) in the LC. To examine further the interactions among dynorphin (DYN), ENK, and CRF in the LC, sections were processed for detection of DYN and CRF or DYN and ENK in rat brain. DYN- and CRF containing axon terminals overlapped noradrenergic dendrites in this region. Dual immunoelectron microscopy showed coexistence of DYN and CRF; 35% of axon terminals containing DYN were also immunoreactive for CRF. In contrast, few axon terminals contained both DYN and ENK. A potential DYN/CRF afferent is the central nucleus of the amygdala (CeA). Dual in situ hybridization showed that, in CeA neurons, 31% of DYN mRNA-positive cells colocalized with CRF mRNA, whereas 53% of CRF mRNA-containing cells colocalized with DYN mRNA. Finally, to determine whether limbic DYN afferents target the LC, the CeA was electrolytically lesioned. Light-level densitometry of DYN labeling in the LC showed a significant decrease in immunoreactivity on the side of the lesion. Taken together, these data indicate that DYN- and CRF-labeled axon terminals, most likely arising from amygdalar sources, are positioned dually to affect LC function, whereas DYN and ENK function in parallel. PMID- 18381636 TI - Long-term evaluation of a calcium phosphate bone cement with carboxymethyl cellulose in a vertebral defect model. AB - We investigated histological and compressive properties of a calcium phosphate bone cement (BoneSource (CPC); Stryker Orthopaedics, Mahwah, New Jersey) plus carboxymethyl cellulose (CMC) using a sheep vertebral bone void model. Bone voids were surgically created in L3 and L5 in each of 40 sheep, and the voids were filled with the cement. Histological and radiographic evaluations were performed on one vertebral body from each animal at either: 0, 3, 6, 12, 24, or 36 months after surgery; mechanical testing was performed on operated and non-operated vertebral bodies from 35 sheep. Undecalcified sections were digitized, and the area of the original defect, new bone formation, empty space, fibrous tissue, and residual cement were quantified with histomorphometry. Decalcified sections were evaluated qualitatively. The cement was biocompatible, extremely osteoconductive and underwent steady resorption and replacement by bone and bone marrow. Histomorphometry showed variations in the rate of cement remodeling among animals in each time group, but on average, at 36 months the original defect area was occupied by approximately 14% bone, 82% cement, and 4% bone marrow. Even in animals that had greater resorption of cement, there was good bone ingrowth with no fibrous tissue. Compressive testing did not reveal a significant difference in the mechanical properties between vertebral bodies augmented with cement and non augmented controls, irrespective of the postoperative time. BoneSource mixed with CMC had adequate osteoconductivity, biocompatibility, and adequate compressive strength. There was variability among animals, but histology suggests that considerable cement was still present in most samples after 36 months. PMID- 18381637 TI - Effect of dual growth factor delivery on chondrogenic differentiation of rabbit marrow mesenchymal stem cells encapsulated in injectable hydrogel composites. AB - An injectable hydrogel composite consisting of oligo(poly(ethylene glycol)fumarate) (OPF) and gelatin microparticles has been developed as a novel carrier system for cells and growth factors. Rabbit marrow mesenchymal stem cells (MSCs) and gelatin microparticles (MPs) loaded with insulin-like growth factor-1 (IGF-1), transforming growth factor-beta1 (TGF-beta1), or a combination of both growth factors were mixed with OPF, a poly(ethylene glycol)-diacrylate crosslinker and the radical initiators ammonium persulfate and N,N,N',N' tetramethylethylenediamine, and then crosslinked at 37 degrees C for 8 min to form hydrogel composites. Hydrogel composites encapsulating rabbit marrow MSCs and blank MPs served as controls. At day 14, confocal fluorescent images of OPF hydrogels showed a strong aggregation of rabbit marrow MSCs when encapsulated with IGF-1-loaded MPs with or without TGF-beta1-loaded MPs. Quantitative RT-PCR results showed that rabbit marrow MSCs encapsulated with MPs loaded with TGF beta1 or both TGF-beta1 and IGF-1 had a significant increase in the expression of chondrocyte-specific genes such as collagen type II and aggrecan at day 14 as compared with the control group. Specifically, samples with both TGF-beta1-loaded MPs and IGF-1-loaded MPs exhibited a 121 +/- 20-fold increase of type II collagen gene expression and a 71 +/- 24-fold increase of aggrecan gene expression after 14 days of in vitro culture as compared with controls at day 0. These results suggest that hydrogel composites based on OPF and gelatin microparticles have great potential as carriers for MSCs and multiple growth factors for cartilage tissue engineering applications. PMID- 18381638 TI - Fibronectin modulates osteoblast behavior on Nitinol. AB - We have previously demonstrated that primary rat osteoclasts behave differently when cultured on austenite and martensite Nitinol. In this study, we coated the two phases of Nitinol with plasma fibronectin and studied if this modifies the proliferation and cell cycle of MC3T3-E1 osteoblasts. The influence of the crystalline structure of Nitinol on the remodeling and conformation of fibronectin was also studied. The results on austenite demonstrated that fibronectin was more strongly remodeled and the cells spread better compared with the martensite phase. Interestingly, the conformation of the protein showed no differences between austenite and martensite. In addition, fibronectin improved cell proliferation in both phases, but the effect of fibronectin coating was stronger on the austenite surface. In addition, in both Nitinol phases, the proportion of cells in the G(1) phase was observed to grow in the presence of fibronectin. This could indicate cell differentiation on Nitinol. PMID- 18381639 TI - Photo-crosslinkable, biomimetic, and thermo-sensitive pluronic grafted hyaluronic acid copolymers for injectable delivery of chondrocytes. AB - Hyaluronic acid (HA) grafted with Pluronic F127 copolymer was used as biomimetic hydrogels for cell delivery. The graft copolymer was synthesized by conjugating amine end-capped Pluronic F127 to carboxylic groups of HA using coupling agents. The synthesized HA-g-Pluronic exhibited thermo-sensitive sol-gel transition behaviors over the temperature range of 20-40 degrees C. HA-g-Pluronic copolymers with vinyl groups were photo-crosslinked to prepare more robust hydrogels for cell cultivation. For improved cellular adhesion and proliferation, cell adhesive peptide (Arg-Gly-Asp (RGD)) was additionally conjugated to the HA backbone. The resultant thermo-sensitive, photocrosslinkable, and RGD modified HA-g-Pluronic copolymers were used to encapsulate and cultivate bovine chondrocytes in vitro. A tissue containing cartilage-like components such as GAG and type II collagen was successfully produced within the hydrogels, indicating that the synthesized HA-g Pluronic copolymers can be potentially used as an injectable cell carrier. PMID- 18381640 TI - Genes, chromosomes and the development of testicular germ cell tumors of adolescents and adults. AB - Testicular germ cell tumors (TGCTs) of adults and adolescents are thought to be derived from primordial germ cells or gonocytes. TGCTs develop postpuberty from precursor lesions known as intratubular germ cell neoplasia undifferentiated. The tumors can be divided into two groups based on their histology and clinical behavior; seminomas resemble primordial germ cells or gonocytes and nonseminomas resemble embryonic or extraembryonic tissues at various stages of differentiation. The most undifferentiated form of nonseminoma, embryonal carcinoma, resembles embryonic stem cells in terms of morphology and expression profiling, both mRNAs and microRNAs. Evidence supports both environmental factors and genetic predisposition underlying the development of TGCTs. Various models of development have been proposed and are discussed. In TGCTs, gain of material from the short arm of chromosome 12 is invariable: genes from this region include the proto-oncogene KRAS, which has activating mutations in approximately 10% of tumors or is frequently overexpressed. A number of different approaches to increase the understanding of the development and progression of TGCTs have highlighted the involvement of KIT, RAS/RAF/MAPK, STAT, and PI3K/AKT signaling. We review the role of these signaling pathways in this process and the potential influence of environmental factors in the development of TGCTs. PMID- 18381641 TI - Secondary genomic rearrangements involving immunoglobulin or MYC loci show similar prevalences in hyperdiploid and nonhyperdiploid myeloma tumors. AB - The pathogenesis of multiple myeloma (MM) is thought to involve at least two pathways, which generate hyperdiploid (HRD) or nonhyperdiploid (NHRD) tumors, respectively. Apart from chromosome content, the two pathways are distinguished by five primary immunoglobulin heavy chain (IGH) rearrangements (4p16, FGFR3, and MMSET; 6p21, CCND3; 11q13, CCND1; 16q23, MAF; 20q12, MAFB) that are present mainly in NHRD tumors. To determine the prevalence and structures of IGH, immunoglobulin (IG) light chain, and MYC genomic rearrangements in MM, we have done comprehensive metaphase fluorescent in situ hybridization analyses on 48 advanced MM tumors and 47 MM cell lines. As expected, the prevalence of the five primary IGH rearrangements was nearly 70% in NHRD tumors, but only 12% in HRD tumors. However, IGH rearrangements not involving one of the five primary partners, and IG light chain rearrangements, have a similar prevalence in HRD and NHRD tumors. In addition, MYC rearrangements, which are thought to be late progression events that sometimes do not involve an IG heavy or light chain locus, also have a similar prevalence in HRD and NHRD tumors. In contrast to the primary IGH rearrangements, which usually are simple balanced translocations, these other IG rearrangements usually have complex structures, as previously described for MYC rearrangements in MM. We conclude that IG light chain and MYC rearrangements, as well as secondary IGH rearrangements, make similar contributions to the progression of both HRD and NHRD MM tumors. PMID- 18381642 TI - Deep brain pulse-generator and lead-extensions: subjective sensations related to measured parameters. AB - This study analyzed subjective sensations caused by DBS pulse-generator and lead extensions in relation with objectively measured parameters. In 50 patients implant-related sensations were evaluated. The pulse-generator mobility was video analyzed. Insufficient lead-extension/pulse-generator tolerability (72%/84%) was documented. Furthermore, 54% of the patients described movement impairments and 48% cosmetic deformity. High body mass index (BMI) was associated with low lead extension related pain (P < 0.001). High generator mobility resulted in high lead extension related pain (P < 0.001). Compared with lead-extension type 7482, type 7495 showed less lead-extension related pain (P = 0.0138), we suppose secondary to surgical tunneling instruments with a larger tip diameter. The lead-extension path with one tissue tunnel for both lead-extensions had 36% lead-extension related pain versus 11% for the path with one single tunnel for each lead extension. Smaller pulse-generators for better cosmetic results, surgical procedures using larger tunneling instruments and one single tunnel for each lead extension would provide better results for patients with BMI <30. PMID- 18381643 TI - Falls and gait disturbances in Huntington's disease. AB - Falls are common in patients with Huntington's disease, but the incidence, falling circumstances and contributing factors have never been examined. We recorded falls in 45 early to midstage Huntington's disease patients, both retrospectively (12 months) and prospectively (3 months). Fall rates were related to relevant baseline measures, including the Unified Huntington's Disease Rating Scale (UHDRS) and quantitative measures of balance (using angular velocity sensors) and gait (using a pressure-sensitive walkway). Balance and gait measures were compared between patients and 27 healthy age-matched controls. Twenty-seven patients (60%) reported two or more falls in the previous year and were classified as fallers. During prospective follow-up 40% reported at least one fall. A high proportion of falls (72.5%) caused minor injuries. Compared to nonfallers, fallers showed significantly higher scores for chorea, bradykinesia and aggression, as well as lower cognitive scores. Compared to controls, Huntington patients had a decreased gait velocity (1.15 m/s versus 1.45 m/s, P < 0.001) and a decreased stride length (1.29 m versus 1.52 m, P < 0.001). These abnormalities were all significantly greater in fallers compared to nonfallers. In addition, fallers had an increased stride length variability and a significantly greater trunk sway in medio-lateral direction compared to nonfallers. We conclude that falls are common in Huntington's disease. Contributing factors include a combination of "motor" deficits (mainly gait bradykinesia, stride variability and chorea, leading to excessive trunk sway), as well as cognitive decline and perhaps behavioral changes. These factors should be considered as future targets for therapies that aim to reduce falls in Huntington's disease. PMID- 18381644 TI - Task-specific craniocervical dystonia. AB - The pathophysiology of dystonia is incompletely understood. Unlike many other focal dystonias, cervical dystonia, a frequent dystonia leading to twisting of the head, does not appear to be related to overuse or acquisition of a demanding motor skill. Here, we report development of task-specific dystonia of the neck muscles in a 67-year-old patient following bilateral traumatic arm amputation at the age of 15. To compensate for the amputation, the patient learned to write with a pen held in his mouth. After several years of practicing this unusual and demanding skill, symptoms of task-specific cervical dystonia (CD) developed. This dystonia later became permanent, and independent of the motor activity that initially triggered the dystonic muscular contractions. This singular case raises the possibility that the pathophysiology of CD may share common elements with that of focal dystonias in different body regions. PMID- 18381646 TI - The Montreal cognitive assessment as a screening tool for cognitive impairment in Parkinson's disease. AB - Cognitive impairment is common in Parkinson's disease (PD) and can occur early in the disease course. No effective screening test exists for detection of early or mild cognitive impairment in PD. We examined the Montreal Cognitive Assessment (MoCA) as a screening tool for cognitive dysfunction in PD. The test-retest intraclass correlation coefficient was 0.79 and the interrater intraclass correlation coefficient was 0.81. The correlation coefficient between the MoCA and a neuropsychologic battery was 0.72. The MoCA is reliable and valid in the PD population and warrants further study as a screening tool for cognitive dysfunction. PMID- 18381645 TI - Childhood-onset restless legs syndrome: clinical and genetic features of 22 families. AB - Restless legs syndrome (RLS) is a sensory-motor disorder that is underdiagnosed in children and often misclassified as attention deficit hyperactivity disorder. Five different gene loci (RLS1-5) and three susceptibility loci have been identified in adult-onset RLS. We included 23 children with RLS (age at onset < or =14 years) from 22 families. In 14 families, we performed linkage and genotype analyses. Of the 23 RLS patients, only seven (30.4%) were admitted for a suspected diagnosis of RLS. Five patients had a retrospectively established onset at an age as early as 1 year. The most frequent complaint in patients were sleep problems (21 of 23; 91%) resulting in fatigue in 14 children (60.9%). Twelve of the 19 tested cases (63.2%) exhibited an index of periodic limb movements in sleep greater than 5. Dopaminergic therapy was successful in 12 of 14 treated patients (85.7%). Family history for RLS was positive in 20 of 23 children (87.0%) and compatible with an autosomal dominant inheritance pattern. Linkage analysis excluded all five loci in two families. A trend for an association at two of the three reported susceptibility regions was observed. RLS symptoms can occur in early childhood. The positive family history suggests a genetic cause in most families with at least one additional RLS gene locus. PMID- 18381647 TI - Parkinson's disease-cognitive rating scale: a new cognitive scale specific for Parkinson's disease. AB - Cognitive defects associated with cortical pathology may be a marker of dementia in Parkinson's disease (PD). There is a need to improve the diagnostic criteria of PD dementia (PDD) and to clarify the cognitive impairment patterns associated with PD. Current neuropsychological batteries designed for PD are focused on fronto-subcortical deficits but are not sensitive for cortical dysfunction. We developed a new scale, the Parkinson's Disease-Cognitive Rating Scale (PD-CRS), that was designed to cover the full spectrum of cognitive defects associated with PD. We prospectively studied 92 PD patients [30 cognitively intact (CogInt), 30 mild cognitive impairment (MCI), 32 PDD] and 61 matched controls who completed the PD-CRS and neuropsychological tests assessing the cognitive domains included in the PD-CRS. Acceptability, construct validity, reliability, and the discriminative properties of the PD-CRS were examined. The PD-CRS included items assessing fronto-subcortical defects and items assessing cortical dysfunction. Construct validity, test-retest and inter-rater reliability of PD-CRS total scores showed an intraclass correlation coefficient >0.70. The PD-CRS showed an excellent test accuracy to diagnose PDD (sensitivity 94%, specificity 94%). The PD-CRS total scores and confrontation naming item scores-assessing "cortical" dysfunction-independently differentiated PDD from non-demented PD. Alternating verbal fluency and delayed verbal memory independently differentiated the MCI group from both controls and CogInt. The PD-CRS appeared to be a reliable and valid PD-specific battery that accurately diagnosed PDD and detected subtle fronto-subcortical deficits. Performance on the PD-CRS showed that PDD is characterized by the addition of cortical dysfunction upon a predominant and progressive fronto-subcortical impairment. PMID- 18381648 TI - Evaluation of a screening instrument for essential tremor. AB - To evaluate a screening instrument for essential tremor (ET) consisting of a seven-item questionnaire and a spiral drawing. A total of 2,448 Danish twins aged 70 years or more and a second sample aged 60 years or more (n = 1,684) from a population-based northern German cross-sectional study (PopGen ET) were screened for ET. Inclusion criteria were a previous diagnosis of ET, a positive answer to two or more questions of the questionnaire or a spiral rating >4 (range of scale 0-9). Three hundred thirteen of 380 positively screened and 321 negatively screened subjects were clinically examined. Definite or probable ET was diagnosed in 104 patients, possible in 86 and other tremors in 98 patients. The sensitivity of the screening instrument was 70.5%, the positive predictive value was 64.9%, the specificity was 68.2%, and the negative predictive value was 73.5%. Tremor severity correlated significantly with higher spiral scores and more positive items. More patients were identified by spiral drawing in all tremor groups. The interrater and intrarater reliability for spirals ranged from 0.7 to 0.8 using intraclass coefficient. A cluster analysis revealed that the questionnaire can be reduced to three items, about uncontrollable tremor in any body part, tremor while drinking or pouring and other family members with tremor, without loosing efficacy. We present an easy to use and reliable screening instrument that is effective to identify patients with ET but not able to exclude patients with other tremor forms. PMID- 18381649 TI - Low LDL cholesterol and increased risk of Parkinson's disease: prospective results from Honolulu-Asia Aging Study. AB - Low-density lipoprotein cholesterol (LDL-C) levels are suggested to be associated inversely with Parkinson's disease (PD). To test the hypothesis that LDL-C levels may increase PD risk, we studied a prospective cohort of 3,233 men (Honolulu-Asia Aging Study) for whom the LDL-C from fasting lipid profiles was obtained during 1991 to 1993. The cohort was followed longitudinally until 2001 for incident Parkinson's cases. During follow-up, 41 men developed PD (18.4/10,000 person years). Although the incidence of PD increased with decreasing LDL-C in a dose dependent manner, the association was only significant for men aged 71 to 75 years. In the latter group, risk of PD declined from 38.5/10,000 person-years in men with LDL-C levels <80 mg/dl to less than 9/10,000 person-years for concentrations that were > or =140 mg/dl. After adjustment for age, smoking, coffee intake, and other factors, the relative odds of PD for men at the 80th versus the 20th percentile of LDL-C (135 vs. 85 mg/dl) was 0.4 (95% confidence interval: 0.2, 0.9). This prospective study supports the hypothesis that low LDL C is associated with an increased risk of PD. Although confirmation is required, the underlying mechanisms may be useful in understanding key aspects of PD. PMID- 18381650 TI - Expression of glutamine synthetase and glutamate dehydrogenase in the latent phase and chronic phase in the kainate model of temporal lobe epilepsy. AB - It has been suggested that astrocytic glutamate release or perturbed glutamate metabolism contributes to the proneness to epileptic seizures. Here we investigated whether astrocytic contents of the major glutamate degrading enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH) decreases on moving from the latent phase (prior to seizures) to the chronic phase (after onset of seizures) in the kainate (KA) model of temporal lobe epilepsy. Western blotting and immunogold analysis of hippocampal formation indicated similar levels of GDH in the latent and chronic phases of KA injected rats and in corresponding controls. In contrast, the level of GS was increased in the latent phase compared with controls, as assessed by Western blots of whole hippocampal formation and subregions. The increase in GS paralleled that of glial fibrillary acidic protein (GFAP). Compared with the latent phase, the chronic phase revealed a lower level of GS (approaching control levels) but an unchanged GFAP content. The decrease in GS from latent to chronic phase was significant in whole hippocampal formation, dentate gyrus and CA3. It is concluded that kainate treated rats show an initial increase in GS, pari passu with the increase in GFAP, and a secondary decrease in GS that is not accompanied by a similar loss of GFAP. In a situation where glutamate catabolism is in high demand the secondary reduction in GS level may be sufficient to contribute to the seizure proneness that develops between the latent and chronic phases. PMID- 18381651 TI - A BAC transgenic mouse model to analyze the function of astroglial SPARCL1 (SC1) in the central nervous system. AB - Extracellular matrix associated Sparc-like 1 (SC1/SPARCL1) can influence the function of astroglial cells in the developing and mature central nervous system (CNS). To examine SC1's significance in the CNS, we generated a BAC transgenic mouse model in which Sc1 is expressed in radial glia and their astrocyte derivatives using the astroglial-specific Blbp (Brain-lipid binding protein; [Feng et al., (1994) Neuron 12:895-908]) regulatory elements. Characterization of these Blbf-Sc1 transgenic mice show elevated Sc1 transcript and protein in an astroglial selective pattern throughout the CNS. This model provides a novel in vivo system for evaluating the role of SC1 in brain development and function, in general, and for understanding SC1's significance in the fate and function of astroglial cells, in particular. PMID- 18381652 TI - PKC-dependent endocytosis of the GLT1 glutamate transporter depends on ubiquitylation of lysines located in a C-terminal cluster. AB - The activity of the main glutamate transporter in the CNS, GLT1, can be regulated by protein kinase C (PKC). It is known that activation of PKC by phorbol esters promotes the clathrin-dependent internalization of the transporter, followed by its lysosomal degradation. However, the molecular mechanisms that link PKC activation and the internalization of GLT1 are not fully understood. In this article, we show that this internalization process is dependent on the ubiquitylation of lysine residues located in the C-terminal tail of GLT1. Exposure to PMA increases the ubiquitylation of GLT1 in transfected cells and in the rat brain, and this ubiquitylated GLT1 accumulates in the intracellular compartment. However, internalization of ubiquitylated GLT1 was blocked with a dominant negative dynamin 2 mutant, indicating that the addition of ubiquitin moieties to the transporter in the membrane precedes its endocytosis. The elimination of lysines from the C-terminus of the transporter (lysines 497, 517, 526, 550, 558, 570, and 573) blocked GLT1 ubiquitylation and endocytosis. However, reintroduction of lysine 517 alone into this mutant was sufficient to restore PMA dependent ubiquitylation and internalization of GLT1. Similarly, reintroduction of lysine 526 restored the endocytosis, while this was only partially recovered after the individual reintroduction of lysines 550 or 570. These data suggest that the activation of PKC induces the ubiquitylation of these C-terminal lysine residues in GLT1 and that this modification mediates the interaction of the transporter with the endocytic machinery. PMID- 18381653 TI - Differential involvement of TNF alpha in hypoxic suppression of astrocyte glutamate transporters. AB - Transporter-mediated glutamate uptake is a principal function of astrocytes. Our previous studies have shown that this process is compromised under hypoxic conditions through the NF-kappaB mediated inhibition of expression of the glutamate transporters EAAT-1 and EAAT-2. Here, we demonstrate that identical conditions of hypoxia (1% O(2), 24 h) lead to a dramatic increase in TNFalpha production from astrocytes without altering their viability. This hypoxia-evoked production of TNFalpha was prevented in the presence of any of three mechanistically distinct NF-kappaB inhibitors. Exogenous application of TNFalpha was without effect on EAAT-1 expression as determined by Western blotting, but mimicked the effects of hypoxia to suppress expression of EAAT-2. Furthermore thalidomide, which prevents TNFalpha production, was without effect on hypoxic suppression of EAAT-1 but prevented hypoxic suppression of EAAT-2. These data indicate that regulation of glutamate transporter expression in astrocytes by hypoxia is subtype specific. Regulation of both EAAT-1 and EAAT-2 is mediated by NF-kappaB, and this transcriptional regulator is also required for increased production of TNFalpha. However, while TNFalpha is essential for hypoxic suppression of EAAT-2, hypoxic modulation of EAAT-1 expression is unaffected by this cytokine. PMID- 18381654 TI - Opioid addiction and pregnancy: perinatal exposure to buprenorphine affects myelination in the developing brain. AB - Buprenorphine is a mu-opioid receptor partial agonist and kappa-opioid receptor antagonist currently on trials for the management of pregnant opioid-dependent addicts. However, little is known about the effects of buprenorphine on brain development. Oligodendrocytes express opioid receptors in a developmentally regulated manner and thus, it is logical to hypothesize that perinatal exposure to buprenorphine could affect myelination. To investigate this possibility, pregnant rats were implanted with minipumps to deliver buprenorphine at 0.3 or 1 mg/kg/day. Analysis of their pups at different postnatal ages indicated that exposure to 0.3 mg/kg/day buprenorphine caused an accelerated and significant increase in the brain expression of all myelin basic protein (MBP) splicing isoforms. In contrast, treatment with the higher dose caused a developmental delay in MBP expression. Examination of corpus callosum at 26-days of age indicated that both buprenorphine doses cause a significant increase in the caliber of the myelinated axons. Surprisingly, these axons have a disproportionately thinner myelin sheath, suggesting alterations at the level of axon-glial interactions. Analysis of myelin associated glycoprotein (MAG) expression and glycosylation indicated that this molecule may play a crucial role in mediating these effects. Co-immunoprecipitation studies also suggested a mechanism involving a MAG-dependent activation of the Src-family tyrosine kinase Fyn. These results support the idea that opioid signaling plays an important role in regulating myelination in vivo and stress the need for further studies investigating potential effects of perinatal buprenorphine exposure on brain development. PMID- 18381655 TI - Adenosine induces hemeoxygenase-1 expression in microglia through the activation of phosphatidylinositol 3-kinase and nuclear factor E2-related factor 2. AB - Adenosine, a purine nucleoside, has been reported to suppress the inflammatory responses of microglia in the brain. However, the underlying mechanisms of its anti-inflammatory action are unclear at present. Here we show that adenosine reduces the increase in intracellular reactive oxygen species (ROS) through expression of an antioxidant enzyme, hemeoxygenase-1 (HO-1). The H(2)O(2)-induced intracellular ROS level was significantly low in microglia pretreated with adenosine for 3-6 h, compared with that in untreated cells. Adenosine induced HO 1 mRNA and protein expression within 3 h, which was maintained for up to 12 h. Nuclear factor E2-related factor 2 (Nrf2), a transcription factor, and phosphatidylinositol 3-kinase (PI3K) and Akt pathways appear to mediate HO-1 expression. In response to adenosine, Nrf2 translocated from the cytosol to nuclei, and bound to the antioxidant response element (ARE). Adenosine enhanced HO-1 promoter activity in an ARE-dependent manner. Moreover, the nucleoside stimulated Akt phosphorylation, and suppressors of PI3K (LY294002 and wortmannin) reduced adenosine-induced HO-1 expression. However, we propose that the effects of adenosine are independent of adenosine receptors, since agonists and antagonists of A1, A2a, and A3 had little effect on the regulation of intracellular ROS and HO-1 expression. Our results collectively suggest that adenosine acts as an endogenous regulator of brain inflammation via modulation of microglial ROS production. PMID- 18381656 TI - Differential neutrophil infiltration contributes to regional differences in brain inflammation in the substantia nigra pars compacta and cortex. AB - Brain inflammation is a suggested risk factor for neurodegenerative disease. Interestingly, severe inflammation in the substantia nigra pars compacta (SNpc) accelerates the onset and progression of Parkinson's disease. In this study, we examined the underlying mechanisms of severe inflammation in the SNpc by comparing the inflammatory process with that in the cortex. In intact brain, the densities of CD11b(+) microglia were similar in the SNpc and cortex. However, lipopolysaccharide injection enhanced the CD11b(+) cell number in the SNpc, but not in the cortex. Previously, we reported that CD11b and myeloperoxidase (MPO) double-positive neutrophils infiltrate the SNpc following LPS injection (GLIA 55:1577-88). Notably, the MPO(+) neutrophil number increased dramatically in the SNpc, but only slightly in the cortex. The extent of neutrophil infiltration appeared to correlate with neuronal damage. We confirmed that loss of neurons in the SNpc was significantly reduced in neutropenic rats versus normal rats following LPS injection. In addition, the densities of astrocytes were much lower in the intact SNpc, compared with the cortex. Furthermore, after LPS injection, damage of endothelial cells and astrocytes, and blood-brain barrier (BBB) permeability was more pronounced in the SNpc. These results collectively suggest that excessive neutrophil infiltration and environmental factors, such as lower astrocyte density and higher BBB permeability, contribute to severe inflammation and neuronal death in the SNpc. PMID- 18381657 TI - Management of obstetrical brachial plexus palsy with early plexus microreconstruction and late muscle transfers. AB - Birth brachial plexus injury usually affects the upper roots. In most cases, spontaneous reinnervation occurs in a variable degree. This aberrant reinnervation leaves characteristic deformities of the shoulder, elbow, forearm, wrist, and hand. Common sequelae are the internal rotation and adduction deformity of the shoulder, elbow flexion contractures, forearm supination deformity, and lack of wrist extension and finger flexion. Nowadays, the strategy in the management of obstetrical brachial plexus palsy focuses in close follow-up of the baby up to 3-6 months and if there are no signs of recovery, microsurgical repair is indicated. Nonetheless, palliative surgery consisting of an ensemble of secondary procedures is used to further improve the overall function of the upper extremity in patients who present late or fail to improve after primary management. These secondary procedures include transfers of free vascularized and neurotized muscles. We present and discuss our experience in treating early and/or late obstetrical palsies utilizing the above-mentioned microsurgical strategy and review the literature on the management of brachial plexus birth palsy. PMID- 18381658 TI - A simple innovation to improve recipient vessel exposure in the axilla during microvascular breast reconstruction with DIEAP flap. PMID- 18381659 TI - Role of timing in assessment of nerve regeneration. AB - Small animal models are indispensable for research on nerve injury and reconstruction, but their superlative regenerative potential may confound experimental interpretation. This study investigated time-dependent neuroregenerative phenomena in rodents. Forty-six Lewis rats were randomized to three nerve allograft groups treated with 2 mg/(kg day) tacrolimus; 5 mg/(kg day) Cyclosporine A; or placebo injection. Nerves were subjected to histomorphometric and walking track analysis at serial time points. Tacrolimus increased fiber density, percent neural tissue, and nerve fiber count and accelerated functional recovery at 40 days, but these differences were undetectable by 70 days. Serial walking track analysis showed a similar pattern of recovery. A "blow-through" effect is observed in rodents whereby an advancing nerve front overcomes an experimental defect given sufficient time, rendering experimental groups indistinguishable at late time points. Selection of validated time points and corroboration in higher animal models are essential prerequisites for the clinical application of basic research on nerve regeneration. PMID- 18381660 TI - Microsurgery and its platform. PMID- 18381661 TI - Repopulation of donor cells from the recipient following extremity graft: studies using the LACZ transgenic rat. AB - Cell traffic from the recipient into the graft, so called graft chimerism, appears to play a major role in graft acceptance. This study was conducted using LacZ transgenic rats to define recipient cell migration into whole-limb grafts. Eighteen hind limb transplants were performed across LacZ+ transgenic and LacZ- rats. LacZ expression was evaluated by X-gal staining. Intense LacZ expression was observed at the interfollicular epidermis in the skin component of LacZ- limb grafts 48 weeks after transplant. Muscle fibers and cartilage showed intense LacZ expression in LacZ+ limb grafts at 48 weeks but no expression in LacZ- grafts. Polymerase chain reaction studies confirmed the presence of the LacZ gene in the bone and skin of LacZ- limb grafts and in the muscle of LacZ+ grafts. Our results demonstrate that there is a gradual increase in repopulation of recipient cells in grafted skin and bone tissues, but muscle and cartilage tissues retain donor cell origin. PMID- 18381662 TI - Overexpression of JKTBP1 induces androgen-independent LNCaP cell proliferation through activation of epidermal growth factor-receptor (EGF-R). AB - Heterogenous nuclear ribonucleoprotein D-like protein (JKTBP) belongs to a new member of hnRNPs. Previous studies implied that JKTBP1 may be associated with the progression of androgen-independent (AI) prostate cancer. In this study, we generated three stable LNCaP cell lines which expressed exogenous JKTBP1. Furthermore, the effect of ectopic JKTBP1 on the proliferation of LNCaP cells and its mechanism was investigated. We originally found that the ectopic JKTBP1 expression resulted in the proliferation of LNCaP cells in an AI way, as well as inducing the upregulated expression of EGF-R and prostate-specific antigen (PSA), but did not influence the expression level of AR. Moreover, AG1478 suppressed the effect of proliferation induced by JKTBP1. In addition, immunohistochemistry showed that JKTBP1 expression was significantly elevated in AI prostate cancer tissues when compared with the androgen-dependent (AD) prostate cancer and benign prostatic hyperplasia. Our data indicated that overexpression of JKTBP1 in LNCaP cells leads to abnormal cell proliferation and may be involved in the process of AD to AI through induction of EGF-R expression. PMID- 18381665 TI - An atomistic branching mechanism for carbon nanotubes: sulfur as the triggering agent. PMID- 18381663 TI - Photodissociation of CH2I2 and subsequent electron transfer in solution. AB - We studied photoinduced reactions of diiodomethane (CH(2)I(2)) upon excitation at 268 nm in acetonitrile and hexane by subpicosecond-nanosecond transient absorption spectroscopy. The transient spectra involve two absorption bands centered at around 400 (intense) and 540 nm (weak). The transients probed over the range 340-740 nm show common time profiles consisting of a fast rise (<200 fs), a fast decay ( approximately 500 fs), and a slow rise. The two fast components were independent of solute concentration, whereas the slow rise became faster (7-50 ps) when the concentration in both solutions was increased. We assigned the fast components to the generation of a CH(2)I radical by direct dissociation of the photoexcited CH(2)I(2) and its disappearance by subsequent primary geminate recombination. The concentration-dependent slow rise produced the absorption bands centered at 400 and 540 nm. The former consists of different time-dependent bands at 385 and 430 nm. The band near 430 nm grew first and was assigned to a charge-transfer (CT) complex, CH(2)I(2) (delta+)I(delta-), formed by a photofragment I atom and the solute CH(2)I(2) molecule. The CT complex is followed by full electron transfer, which then develops the band of the ion pair CH(2)I(2) (+)I(-) at 385 nm on the picosecond timescale. On the nanosecond scale, I(3) (-) was generated after decay of the ion pair. The reaction scheme and kinetics were elucidated by the time-resolved absorption spectra and the reaction rate equations. We ascribed concentration-dependent dynamics to the CT-complex formation in pre-existing aggregates of CH(2)I(2) and analyzed how solutes are aggregated at a given bulk concentration by evaluating a relative local concentration. Whereas the local concentration in hexane monotonically increased as a function of the bulk concentration, that in acetonitrile gradually became saturated. The number of CH(2)I(2) molecules that can participate in CT-complex formation has an upper limit that depends on the size of aggregation or spatial restriction in the neighboring region of the initially photoexcited CH(2)I(2). Such conditions were achieved at lower concentrations in acetonitrile than in hexane. PMID- 18381673 TI - Efficacy of the pyrethroid alpha-cypermethrin against Bactrocera oleae populations from Greece, and improved diagnostic for an iAChE mutation. AB - BACKGROUND: The most important pest of olive orchards worldwide is the olive fruit fly Bactrocera oleae (Gmelin). Its control in Greece has been based on organophosphates (OPs), but their intense use has led to the development of resistance. A test previously developed to monitor the trait may not be as robust as originally thought. The pyrethroid alpha-cypermethrin has recently been registered for bait sprays, as an alternative to OPs. RESULTS: The susceptibility of 20 B. oleae populations to alpha-cypermethrin was examined. Variation was observed in their response, with LD(50) ranging from 0.14 to 3.28 ng insect(-1) and resistance factors from 2.3 to 54.7. Resistance mechanisms were investigated. Cytochrome P450 monoxygenase activities showed an association with resistance. Sequences in the domain IIS4-IIS6 of the B. oleae para-type sodium channel were also analysed, but no resistance-associated mutations were identified. Finally, a novel diagnostic assay able to reliably monitor the frequency of the iAChE G488S resistance mutation was developed. CONCLUSION: This is the first attempt to evaluate the efficacy of alpha-cypermethrin against B. oleae from Greece. Data showed that it can be used effectively, but also highlighted the importance of continuous monitoring. The IIS4-IIS6 sodium channel region is the default area in which to look for resistance mutations if target-site resistance to pyrethroids arises. The application of the novel iAChE molecular diagnostic may facilitate the introduction of pyrethroids alongside OPs currently in use. PMID- 18381674 TI - Complexation study of midazolam hydrochloride with beta-cyclodextrin: NMR spectroscopic study in solution. AB - (1)H NMR spectroscopic study of midazolam hydrochloride (MDL), beta-cyclodextrin (beta-CD) and their mixtures confirmed the formation of beta-CD-MDL inclusion complex in aqueous solution. The stoichiometry of the complexes was determined by Scott's method to be 1:1, and the association constant (K(a)) was calculated to be 108 M(-1). It was confirmed on the basis of 2D ROESY spectral data that only a fluorine-substituted aromatic ring acted as guest in complexation. Most of the aromatic signals of MDL exhibited induced shift changes as well as splitting, in the presence of beta-CD, indicating chiral differentiation of MDL by beta-CD. PMID- 18381675 TI - A unified modeling framework for metabonomic profile development and covariate selection for acute trauma subjects. AB - Acute trauma is often associated with progressive deterioration of multiple organ systems in humans and is the leading cause of death in trauma care units. Identification of specific organ failure in a non-invasive manner and the contribution of different demographic factors on the casual progression of acute trauma are of supreme interests for successful diagnosis, prognosis or monitoring of trauma status. Recently, electrospray ionization and matrix-assisted laser desorption time-of-flight mass spectrometry have been used to identify biomarkers in both proteomics and metabonomics studies. Data sets generated from mass spectrometers in such studies are generally very large in size and thus require the use of sophisticated statistical techniques to glean useful information. In a recent development, Ghosh et al. (BMC Bioinformatics 2008; 9:38) suggested a unified semiparametric approach to distinguish urinary metabolic profiles in a group of traumatic subjects from those of a control group consisting of normal individuals. In this study we have extended their approach by combining available covariate information in the development of metabonomic profile of acute trauma. We have shown that age is a statistically significant covariate across trauma and control group, thus pointing out the fact that prognosis of trauma may be acutely linked with subjects' age. PMID- 18381676 TI - Modeling epilepsy disparities among ethnic groups in Philadelphia, PA. AB - The Centers for Disease Control and Prevention defined epilepsy as an emerging public health issue in a recent report and emphasized the importance of epilepsy studies in minorities and people of low socioeconomic status. Previous research has suggested that the incidence rate for epilepsy is positively associated with various measures of social and economic disadvantage. In response, we utilize hierarchical Bayesian models to analyze health disparities in epilepsy and seizure risks among multiple ethnicities in the city of Philadelphia, Pennsylvania. The goals of the analysis are to highlight any overall significant disparities in epilepsy risks between the populations of Caucasians, African Americans, and Hispanics in the study area during the years 2002--2004 and to visualize the spatial pattern of epilepsy risks by ethnicity to indicate where certain ethnic populations were most adversely affected by epilepsy within the study area. Results of the Bayesian model indicate that Hispanics have the highest epilepsy risk overall, followed by African Americans, and then Caucasians. There are significant increases in relative risk for both African Americans and Hispanics when compared with Caucasians, as indicated by the posterior mean estimates of 2.09 with a 95 per cent credible interval of (1.67, 2.62) for African Americans and 2.97 with a 95 per cent credible interval of (2.37, 3.71) for Hispanics. Results also demonstrate that using a Bayesian analysis in combination with geographic information system (GIS) technology can reveal spatial patterns in patient data and highlight areas of disparity in epilepsy risk among subgroups of the population. PMID- 18381677 TI - The grape and wine constituent piceatannol inhibits proliferation of human bladder cancer cells via blocking cell cycle progression and inducing Fas/membrane bound Fas ligand-mediated apoptotic pathway. AB - Piceatannol (3,3',4,5'-Tetrahydroxy-trans-stilbene) is a polyphenol present in grapes and wine. Piceatannol is a protein kinase inhibitor that modifies multiple cellular targets exerting immunosuppressive, antileukemic, and antitumorigenic activities in several cell lines and animal models. In this study, the antiproliferative activity of piceatannol was investigated. The results showed that piceatannol inhibited the proliferation of T24 and HT1376 human bladder cancer cells by blocking cell cycle progression in the G0/G1 phase and inducing apoptosis. ELISA showed that the G0/G1 phase arrest is due to an increased in the expression of p21/WAF1. An enhancement in Fas/APO-1 and membrane-bound Fas ligand (mFasL) might be responsible for the apoptotic effect induced by piceatannol. Our study reports the novel finding, that the induction of p21/WAF1 and activity of the Fas/mFasL apoptotic system may participate in the antiproliferative activity of piceatannol in T24 and HT1376 cells. PMID- 18381678 TI - In vitro and in vivo evaluation of anticancer actions of natural and synthetic vitamin E forms. AB - The goal of these studies was to investigate the potential anticancer properties of two naturally occurring plant sources and two manufactured synthetic forms of vitamin E, i. e., RRR-alpha-tocopherol (alphaT), RRR-gamma-tocopherol (gammaT), all-rac-alpha-tocopherol (all-rac-alphaT), and all-rac-alpha-tocopheryl acetate (all-rac-alphaTAc) in breast cancer models. Vitamin E compounds were evaluated in vitro for inhibition of colony formation and induction of apoptosis in human MDA MB-435 and MCF-7 breast cancer cells and murine 66cl-4 mammary cancer cells and in vivo for ability to reduce tumor growth and lung and lymph node metastases using the transplantable syngeneic BALB/c mouse 66cl-4-GFP mammary cancer model. gammaT inhibited colony formation and induced apoptosis in all three cancer cell lines. alphaT and all-rac-alphaT were less effective and all-rac-alphaTAc was ineffective. gammaT-induced apoptosis was correlated with activation of caspases 8 and -9 and down-regulation of protein expression of c-FLIP and survivin. In vivo study 1 analyses showed that all-rac-alphaT and all-rac-alphaTAc significantly inhibited tumor growth and inhibited both visible and microscopic size lung metastases. In vivo study 2 analyses showed that alphaT and gammaT reduced tumor growth, but only gammaT reduced tumor growth significantly in comparison to control. In conclusion, synthetic, but not natural, vitamin E exhibits promising anti-cancer properties in vivo. PMID- 18381679 TI - Absence of 2'-deoxyguanosine-carbon 8-bound ochratoxin A adduct in rat kidney DNA monitored by isotope dilution LC-MS/MS. AB - The contribution of DNA adduct formation in the carcinogenic action of the mycotoxin ochratoxin A (OTA) has been subject to much debate. Recently, a carbon bonded ochratoxin A-2'-deoxyguanosine adduct (dGuoOTA) formed by photochemical reaction in vitro has been shown by 32P-postlabeling/TLC to comigrate with a spot detected in DNA isolated from rat and pig kidney following exposure to OTA. Considering the large body of evidence arguing against covalent DNA binding of OTA and the poor resolution and specificity of postlabeling analysis, we developed a stable isotope dilution LC-MS/MS method to analyze dGuoOTA in kidney DNA isolated from rats treated with OTA. dGuoOTA and nitrogen-15-labeled dGuoOTA (15N(5)-dGuoOTA) were prepared by photoirradiation of OTA in the presence of dGuo or nitrogen-15-labeled dGuo. Conditions for DNA hydrolysis were optimized using a synthetic oligonucleotide containing dGuoOTA to ensure complete release of dGuoOTA. The LOD of the method (S/N > 3) was 10 fmol dGuoOTA on-column. However, dGuoOTA was not detected in DNA samples isolated from male F344 rats treated with OTA for up to 90 days at doses known to cause renal tumor formation. Detection limits, calculated for each individual sample based on the absolute LOD and the amount of DNA injected, were as low as 3.5 dGuoOTA/10(9) nucleotides. These data are consistent with previous results showing lack of DNA adduct formation by OTA and demonstrate that dGuoOTA is not formed in biologically relevant amounts under physiological conditions in vivo. PMID- 18381682 TI - Effect of column temperature on enantioseparation of dihydropyrimidinones using alcohol solvated Chiralpak AS and AS-H columns. AB - In this study we report the application of column temperature programs to compare the chromatographic behaviors of four commercially available dihydropyrimidinone (DHP) compounds on Chiralpak AS and AS-H phases under different alcohol solvation conditions such as 1-propanol (1-PrOH), 2-methyl-1-PrOH (2-Me-1-PrOH), 1-butanol (1-BuOH), 2-BuOH, and tert-BuOH (t-BuOH) in n-hexane (n-Hex). It was observed that the apparent retention factors of the DHP compounds on the AS phase in t BuOH/n-Hex mobile phase had the largest reduction (>27%) among the mobile phases studied after a heating and cooling temperature cycle (10 to 50 back to 10 degrees C). However, the average reduction of the apparent retention factors of the compounds on t-BuOH solvated AS-H was less than 4%. No clear trend of changes in the apparent retention factors on AS and AS-H columns was observed in other alcohol modified mobile phases. Step-temperature programs showed that alcohol solvated AS phase had larger changes in the apparent retention factors with time than those of AS-H phase at the elevated temperature step (50 degrees C). Both t BuOH solvated AS and AS-H phases showed kinetic behaviors in chromatographic processes at the elevated temperature. PMID- 18381683 TI - Mixture design optimization of extraction and mobile phase media for fingerprint analysis of Bauhinia variegata L. AB - Two statistical mixture designs were used to optimize the proportions of solvents used in both the extraction medium and the reversed liquid chromatographic mobile phase to improve the quality of chromatographic fingerprints of Bauhinia variegata L extracts. For modeling, the number of peaks was used as a measure of fingerprint information. Three mobile phases, each with a chromatographic strength of two, gave good results. A methanol/water (77:23 v/v) mixture resulted in 17 peaks in the chromatographic fingerprint whereas acetonitrile/water (64.5:35.5 v/v) and methanol/acetonitrile/water (35:35:30 v/v/v) mixtures resulted in 18 and 20 peaks, respectively. The corresponding optimum solvent compositions to extract chemical substances for these three mobile phases were ethanol/acetone (25:75 v/v/v) and dichloromethane/acetone (70:30 v/v) mixtures, and pure dichloromethane, respectively. The mixture designs are useful for understanding the influence of different solvents on the strengths of the extraction medium and the mobile phase. PMID- 18381684 TI - Fast-GC-conventional quadrupole mass spectrometry in essential oil analysis. AB - This study reports on the compatibility of a conventional quadrupole MS (qMS) as detector for Fast-GC in terms of separation, identification, and quantitation when applied to the analysis of peppermint essential oil (e.o.), adopted as representative of this field. The influence of Fast-GC carried out on a 10 mx0.1 mm id narrow bore column with temperature programmes from 20 to 60 degrees C/min on the results of a qMS in total ion chromatograms (TIC) at different speeds (from 999 and 11 111 amu/s) and SIM modes was evaluated on ten differently abundant components characterizing peppermint e.o. Separation measure (S), peak capacity (n), and half height peak width were taken as separation parameters; match quality, number of scans per peak (NP), spectral skewing, and TIC area repeatability were used for identification. Quantitation was in SIM mode and NP, dwell time, SIM area repeatability and calibration curves, LOD, and LOQ of the selected components were measured. The results show that the peppermint e.o. markers can successfully be analysed qualitatively and quantitatively by F-GC-qMS up to temperature programmes of 60 degrees /min provided that a suitable scan speed is applied. Fast-GC-qMS reduces analysis time by a factor greater than ten and gives results that are qualitatively reliable and quantitatively comparable to those obtained by conventional GC-qMS. PMID- 18381685 TI - Lung inflammation and thrombogenic responses in a time course study of Csb mice exposed to ozone. AB - Ozone is a well-known oxidant air pollutant, inhalation of which can result in oxidative stress, and lead to pulmonary inflammation. The aim of this study was to evaluate the time-course events after a single ozone exposure in transcription coupled repair defective Csb and wild type mice. Mice were exposed for 3 h to 2 ppm ozone and biological parameters related to oxidative stress and inflammation were examined in the lungs at 0, 4, 9, 24 and 48 h after exposure. In addition the procoagulant and thrombomodulin activities were explored by a combination of assays for tissue factor and thrombin generation. This study revealed a significant biological response to ozone, for both Csb and wild type mice. The onset of inflammation in Csb mice, as indicated by an increase in interleukin-6, tumor necrosis factor-alpha and total cell influx, occurred earlier compared with those seen in wild type mice. On the other hand, Csb mice showed a delayed antioxidant reaction compared with wild type mice. Both genotypes developed a procoagulant reaction characterized by a stably increased tissue factor activity and a progressive increase in thrombin generation after 2 days. These experiments have shown that ozone, a well-known toxic substance from the environment, induces not only inflammation, but also procoagulant reactions in the lungs of mice. These results have implications for understanding the systemic effects induced by oxidant air pollutants. PMID- 18381686 TI - Volatile compounds from the integument of White Leghorn Chickens (Gallus gallus domesticus L.): candidate attractants of ornithophilic mosquito species. AB - Candidate kairomones of ornithophilic mosquito species are reported from GC/MS analysis of compounds from the skin on the back, the feet, and feathers from the back of White Leghorn chickens (Gallus gallus domesticus). Hexane and ether extracts of chicken feathers differ significantly in attractiveness of Culex spp. mosquitoes. The active (hexane) extracts contained an abundance of alcohols, ketones, and diones. The inactive (ether) extracts contained aldehydes, which also were in the hexane extracts. Analysis of hexane extracts from chicken feet, skin, and feathers demonstrated the qualitative similarity in the compounds collected with subtle differences observed in the quantitative amounts of these compounds. Aldehydes and carboxylic acids were detected in a concentrated ether extract of feathers in quantitatively similar ratios within each compound class for the corresponding series of C(6)-C(9 )aldehydes and acids. PMID- 18381687 TI - Role of phospholipase A2 and tyrosine kinase in Clostridium difficile toxin A induced disruption of epithelial integrity, histologic inflammatory damage and intestinal secretion. AB - Clostridium difficile-associated disease causes diarrhea to fulminant colitis and death. We investigated the role of phospholipase A2 (PLA2) inhibitors, aristolochic acid (AA), bromophenacyl bromide (BPB) and quinacrine (QUIN) on the C. difficile toxin A-induced disruption of epithelial integrity, histologic inflammatory damage and intestinal secretion. Toxin A caused severe hemorrhagic and inflammatory fluid secretion at 6-8 h in rabbit ileal segments, an effect that was significantly inhibited by QUIN (71%, P < 0.01), AA (87%, P < 0.000l) or by BPB (51%, P < 0.01). The secretory effect of toxin A was also inhibited in segments adjacent to those with AA (89%, P < 0.01). Furthermore, QUIN or AA substantially reduced the histologic damage seen after 6-8 h in rabbit ileal segments. The cyclooxygenase inhibitor, indomethacin, also significantly inhibited (96%; n = 6) the secretory effects of toxin A in ligated rabbit intestinal segments. The destruction by toxin A of F-actin at the tight junctions of T-84 cell monolayers was not inhibited by AA or BPB. AA or QUIN had no effect on the T-84 cell tissue resistance reduction over 8-24 h after toxin A exposure. All the inhibitors were shown to be effective in the doses administered direct in ileal loops to inhibit PLA2 activity. The data suggest that PLA2 is involved in the major pathway of toxin A-induced histologic inflammatory damage and hemorrhagic fluid secretion. PMID- 18381688 TI - Analysis of amino acids in human vascular endothelial (ECV-304) cells by microchip electrophoresis with fluorescence detection. AB - A rapid and sensitive method was developed for the analysis of amino acids by microchip electrophoresis with Hg-lamp excitation fluorescence detection. Fluorescein-isothiocyanate (FITC) was chosen to estimate the sensitivity of this system, and the detection limit (S/N = 3) with FITC was 1.7 nM, which showed that the system was sensitive as well as simple. Two derivatizing agents, FITC and ortho-phthalaldehyde (OPA) were employed to label amino acids and were compared in the same fluorescence detection system with an Hg lamp as the excitation source. The separation parameters were optimized in detail. Optimum separation of OPA-labeled amino acids was obtained in less than 200 s with 20 mM borate buffer (pH 9.0) containing 20% acetonitrile and 10 mM beta-cyclodextrin. Detection limits for amino acids (alanine (Ala), taurine (Tau), glycine (Gly), glutamic acid (Glu), and aspartic acid (Asp)) of 0.38-1.0 muM were achieved. The method was successfully applied to analysis of amino acids in human vascular endothelial cells (ECV-304). The average amount of amino acids in single ECV-304 cells is estimated to be 5.84 fmol for Ala, 2.78 fmol for Tau, 1.15 fmol for Gly, 3.10 fmol for Glu, and 1.30 fmol for Asp. PMID- 18381689 TI - The cut-off levels of CD23 expression in the differential diagnosis of MCL and CLL. AB - Flow cytometric analysis of CD23 expression in CD5-positive B-cells is a widely applied method in the differential diagnosis of chronic lymphocytic leukaemia (CLL) and mantle cell lymphoma (MCL). According to the most accepted criteria, the leukaemic cell population is CD19/CD5/CD23 triple positive in CLL but CD23 negative in MCL. Recently, several groups have reported CD23-positive MCL cases; however, these studies mostly analysed only CD23 positivity but not intensity. To determine the role and the cut-off levels of CD23 positivity and intensity in the differential diagnosis of CLL and MCL, 26 cases of MCL and 84 cases of CLL were compared using flow cytometric analysis. Our results suggest that high values of CD23 positivity (>92.5%) and/or high fluorescence intensity (>44.5 mean fluorescence intensity (MFI)) of CD23 are related to CLL, whereas low CD23 positivity (<30%) is related to MCL. However, cases with intermediate CD23 positivity (between 30 and 92.5%) and lower intensity (<44.5 MFI) can either belong to CLL or MCL. In these cases, additional tests such as FISH analysis of the translocation t(11;14) or immunohistochemical detection of cyclin D1 overexpression are required to differentiate CLL from MCL. PMID- 18381690 TI - Screening of anti-platelet aggregation agents from Panax notoginseng using human platelet extraction and HPLC-DAD-ESI-MS/MS. AB - Root of Panax notoginseng (Sanqi in Chinese) is a highly valued and commonly used Chinese medicine. It has been widely used for treatment of cardio- and cerebro vascular diseases. In this study, a method involving human platelet extraction and HPLC-DAD-ESI-MS/MS was developed for screening potential anti-platelet aggregation agents in Sanqi. Five compounds which could interact with human platelets were found, and four were identified as adenosine, guanosine, ginsenoside Rh1, and ginsenoside F1, respectively. The effects on rabbit platelet aggregation induced by arachidonic acid, adenosine diphosphate, and thrombin were also investigated in vitro. The results showed that the nucleosides adenosine and guanosine mainly contributed to the anti-platelet aggregation of Sanqi. The data suggest that human platelet extraction combined with HPLC-DAD-ESI-MS/MS is a useful method for screening anti-platelet aggregation agents from Chinese medicines. PMID- 18381691 TI - Are occupational exposure limits becoming more alike within the European Union? AB - The occupational exposure limits (OELs) established by seven different national regulatory agencies of EU member states are compared with those of the European Commission (EC). The comparison concerned: (1) what chemicals have been selected, (2) the average level of exposure limits for all chemicals, and (3) the similarity between the OELs of different EU member states and the OELs recommended by the European Commission. The average level of the exposure limits has declined during the past 10 years in four of the five countries in our study for which historical data were available to us. Poland has not changed its level noticeably and Germany has increased it. Since the first list of indicative OELs was established by the EC, a few of the EU exposure limits have been lowered. The similarity index indicates that the exposure limits of EU member states are converging towards the European Commission's recommended OELs. Still, the average level of OELs differs between organizations--the Estonian OELs are on average 35% higher than the Polish OELs. PMID- 18381692 TI - 6-Oxy-(acetyl ethylenediamine) fluorescein, a novel fluorescent derivatization reagent for carboxylic acids and its application in HPLC. AB - A novel fluorescent derivatization reagent for carboxylic acids, 6-oxy-(acetyl ethylenediamine) fluorescein (AEF), was well designed, synthesized, and applied to HPLC. The derivatization reaction with 12 fatty acids, including n-valeric acid (C5), n-hexanoic acid (C6), n-heptanoic acid (C7), n-octanoic acid (C8), n nonanoic acid (C9), n-decanoic acid (C10), lauric acid (C12), myristic acid (C14), palmitic acid (C16), stearic acid (C18), oleic acid (C18:1), and linoleic acid (C18:2), was completed at 55 degrees C within 40 min. The derivatives of fatty acids were separated on a C18 RP column and detected by fluorescence detection. The LODs attained were 0.4-1.2 nM (S/N of 3). It has been demonstrated that AEF is a prominent derivatization reagent for carboxylic acids which is suitable for HPLC. PMID- 18381693 TI - Preparation of nitrocellulose (NC) immuno-affinity membrane for purification of rAPC antibody. AB - In this study, recombinant allophycocyanin (rAPC) with a purity of 98% was transferred from a gel to a nitrocellulose (NC) membrane to develop a simple and efficient immuno-affinity membrane. Atomic force microscopy (AFM) was used to investigate the surface topography of the affinity membrane and its characterization indicated that rAPC easily forms trimers or hexamers on the membrane surface on use of the given transfer method. The hydrodynamic radius (R(h)) of the rAPC aggregation was equal to 103 nm or 365 nm according to dynamic light scattering (DLS), which was in agreement with the result obtained by AFM. Based on the specific immunological reaction of antigen and antibody, anti-APC antibodies were purified from rabbit polyclonal serum in a single step. The amount of absorbed antibody was 5.79 mg/g membrane according to analysis by ELISA methods. The purity of antibodies was up to 98% according to SDS-PAGE. The adsorption-desorption cycle of rAPC was repeated six times using the same immuno affinity membrane, and there was no significant loss in adsorption capacity. The method provides a novel and efficient immunological affinity membrane for the purification of antibodies. PMID- 18381697 TI - Editor Joseph Pesek meets Professor Walter Jennings. Interview by Joseph Pesek. PMID- 18381698 TI - Instability of hexane-acetonitrile mobile phases used for the chromatographic analysis of triacylglycerides. AB - Comprehensive 2-D LC is an emerging separation technique that has seen a rapid increase in applications in the last decade. The technique has been applied for the separation of numerous complex mixtures including triacylglycerides (TAG). Determination of TAG in food products such as rice, palm, and canola oils have been previously described and the technique of choice utilizes a silver-modified silica column with hexane-ACN as the mobile phase. Repeated retention time inconsistencies were experienced in our studies when this mobile phase was applied to the separation of natural and synthetic mixtures containing TAG. The present report summarizes a study performed to determine the relative stability of ACN, propionitrile (PCN), and butyronitrile (BCN) at concentrations ranging from 0.43 to 2.8% in hexane and heptane. The data obtained suggest that unless evaporative loss of the mobile phase is prevented, TAG retention time irreproducibility can be significant when using mobile-phase mixtures prepared with ACN or PCN. BCN should be used as the solvent modifier in cases where evaporation cannot be prevented. PMID- 18381699 TI - Preparation and HPLC evaluation of a new 1,3-alternate 25,27-bis-[p chlorobenzyloxy]-26,28-bis-[3-propyloxy]-calix[4]arene silica bonded stationary phase. AB - The 1,3-alternate 25,27-bis-[p-chlorobenzyloxy]-26,28-bis-[3-propyloxy] calix[4]arene-bonded silica gel stationary phase was synthesized, structurally characterized, and used as a selector in high performance liquid chromatography. Selectivity studies on that phase used aromatic positional isomers, alkylbenzenes, polynuclear aromatic hydrocarbons, sulfonamides, and non-steroidal anti-inflammatory drugs as analytes. The effects of organic modifier content and pH of the mobile phase on retention and selectivity of selected aromatic positional isomers were studied. Selectivity comparisons of the novel phase vs. 1,3-alternate 25,27-di-[benzyloxy]-26,28-bis-[3-propyloxy]-calix[4]arene phase and commercially available RP-Phenyl phases were performed. The retention mechanism was also discussed. The results indicated that the calixarene stationary phase behaves like a reversed-phase packing; however, other retention mechanisms seem to be involved in the separation process. PMID- 18381700 TI - Effect of temperature on the chromatographic retention of ionizable compounds. III. Modeling retention of pharmaceuticals as a function of eluent pH and column temperature in RPLC. AB - We propose a general simple equation for accurately predicting the retention factors of ionizable compounds upon simultaneous changes in mobile phase pH and column temperature at a given hydroorganic solvent composition. Only four independent experiments provide the input data: retention factors measured in two pH buffered mobile phases at extreme acidic and basic pH values (e. g., at least +/- 2 pH units far from the analyte pK(a)) and at two column temperatures. The equations, derived from the basic thermodynamics of the acid-base equilibria, additionally require the knowledge of the solute pK(a )and enthalpies of acid base dissociation of both the solute and the buffer components in the hydroorganic solvent mixture. The performance of the predictive model is corroborated with the comparison between theoretical and experimental retention factors of several weak acids and bases of important pharmacological activity, in mobile phases containing different buffer solutions prepared in 25% w/w ACN in water and at several temperatures. PMID- 18381701 TI - Enantioselective HPLC resolution of synthetic intermediates of armodafinil and related substances. AB - Armodafinil is a unique psychostimulant recently approved by the US Food and Drug Administration for the treatment of narcolepsy. The chromatographic resolution of its chiral intermediates including related substances in the total synthesis of armodafinil was studied on polysaccharide-based stationary phases, viz. cellulose tris-(3,5-dimethylphenylcarbamate) (Chiralcel OD-H) and amylose tris-(3,5 dimethylphenylcarbamate) (Chiralpak AD-H) by HPLC. The effects of 1-propanol, 2 propanol, ethanol, and trifluoroacetic acid added to the mobile phase and of column temperature on resolution were studied. A good separation was achieved on cellulose-based Chiralcel OD-H column compared to amylose-based Chiralpak AD-H. The effects of structural features of the solutes and solvents on discrimination between the enantiomers were examined. Baseline separation with R(s) >1.38 was obtained using a mobile phase containing n-hexane-ethanol-TFA (75:25:0.15 v/v/v). Detection was carried out at 225 nm with photodiode array detector while identification of enantiomers was accomplished by a polarimetric detector connected in series. The method was found to be suitable not only for process development of armodafinil but also for determination of the enantiomeric purity of bulk drugs and pharmaceuticals. PMID- 18381702 TI - From healthy start to hurricane Katrina: using GIS to eliminate disparities in perinatal health. AB - This paper provides a summary of the invited talk at the 2007 CDC & ATSDR 11th Biennial Symposium on Statistical Methods conference in which a university-non profit collaboration targeted the elimination of racial disparities in perinatal health with the use of a Geographic Information System (GIS). This program will be described in four temporal stages; the pre-program early years (1999--2001) where the health burden is defined, leading to the Healthy Start years (2001- 2005), in which spatial analyses, methods to effectively disseminate GIS results, the creation of the Baton Rouge Healthy Start database, and a move toward a conceptual goal of creating a holistic neighborhood GIS-health model are all described. The Katrina years (September 2005--early 2006) portrays the impact of the disaster and how the collaboration changed as resources from both were directed toward both response and recovery. The final section of the paper, the Post-Katrina years (early 2006 and ongoing) describes how the health landscape of Louisiana, including Baton Rouge as well as New Orleans, has worsened after the storms. An argument is made that the relationships and GIS structure developed during the collaboration's pre-Katrina years, even though stretched, provide the flexibility to analyze and cope with a Katrina-type shock to the system. PMID- 18381703 TI - Relevance of clinical trials in acute myeloid leukaemia. AB - Patients with newly diagnosed acute myeloid leukaemia (AML) are increasingly being enrolled in clinical trials sponsored by pharmaceutical companies or the National Cancer Institute. These trials routinely exclude patients who are less likely to respond (LLTR), e.g. those with Zubrod performance status >2 and levels of bilirubin or creatinine >/=2.0 mg/dL. Here we examine rates of enrollment of LLTR patients in clinical trials over the past 16 years. Overall, 2323 adults with newly diagnosed AML (excluding acute promyelocytic leukaemia) were registered on clinical trials from 1991 to 2006. LLTR patients constituted a significantly smaller proportion of the patients enrolled from 1999 to 2006 than from 1991 to 1998 (p < 0.0001, considering all patients and patients 60 years or older). While 54% of patients considered 'more likely to respond' (MLTR) (i.e. those with performance status <3, bilirubin <2.0 mg/dL and creatinine <2.0 mg/dL) were enrolled in these studies from 1999 to 2006, only 36% of LLTR patients were enrolled during this period (p < 0.0001). Our results suggest that newer clinical trials may be less applicable to LLTR patients than previous trials. There is a need for clinical trials specific to the LLTR population, the group most in need of novel therapies. PMID- 18381704 TI - Caspase polymorphisms and genetic susceptibility to multiple myeloma. AB - Multiple myeloma is a haematological malignency, characterized by clonal expansion of plasma cells. However, little is known about the cause of multiple myeloma. Cancer cells must avoid apoptosis to ensure unregulated tumour formation and growth. The highly conserved caspase cascade is essential to the regulation of the apoptotic pathway. To examine if five single nucleotide polymorphisms (SNPs) in four caspase genes [CASP3 Ex8-280 C > A (rs6948), CASP3 Ex8 + 567 T > C (rs1049216), CASP8 Ex14-271 A > T (rs13113), CASP9 Ex5 + 32 G > A (rs1052576), CASP10 Ex3-171 A > G (rs39001150)] alter multiple myeloma risk, we conducted a population-based case-control study of women (128 cases; 516 controls) in Connecticut. Compared to individuals with the TT genotype of CASP3 Ex8 + 567 T > C, subjects with the CC genotype had a five-fold decreased risk of multiple myeloma (odds ratio (OR)(CC) = 0.2, 95% confidence interval (CI) = 0.0-1.0). Further, individuals with the AG and AA genotypes of CASP9 Ex5 + 32 G > A also experienced a decreased risk of multiple myeloma (OR(AG) = 0.8, 95% CI = 0.5-1.3; OR(AA) = 0.5, 95% CI = 0.3-0.9; p-trend = 0.02). While no previous study has evaluated the association between caspase genes and multiple myeloma, studies have found associations with lung, breast, esophageal, gastric, colorectal and cervical cancers. Our parallel study of non-Hodgkin lymphoma, which utilized the same controls, found strong evidence that caspase genes play a key role in lymphogenesis. The protective associations observed in two key caspase genes suggest that genetic variation in CASP genes may play an important role in the aetiology of multiple myeloma. PMID- 18381705 TI - The complex formation of tetracyclohexylammonium C1-resorcinarene with various guests - an electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry study. AB - The complex formation of a tetraammonium C1-resorcinarene (R+4HCl) was studied using electrospray ionization Fourier transform ion cyclotron resonance (ESI FTICR) mass spectrometry. Although R+4HCl easily loses its counter ions in the ESI process, a neutral self-assembled structure with an intramolecular circular hydrogen-bonded 16-membered -N(+)-H ... X(-) ... H-N(+)- array with ammonium ion as the charge-giving species was observed in the gas phase. In addition to chloride, several other counter ions were also studied. The size and basicity of the counter ion as well as the size of the charge-giving cation strongly affected the gas-phase stability of the self-assembled system. H/D exchange experiments showed that the ammonium substituents in the apical position of R affect the hydrogen-bonding system in the resorcinarene. The complexation of the saturated dicarboxylic acids was found to depend on the length of the carbon chain. The rigidity of the molecular skeleton of the acid improved the complexation considerably. The orientation and position of the carboxylic groups also had an effect on the complexation and consequently enabled stereochemical differentiation of the acids. Mass spectrometric observations were supported by theoretical calculations. PMID- 18381706 TI - Factors affecting enrollment in literacy studies for English- and Spanish speaking cancer patients. AB - BACKGROUND: Study participation bias can affect inferences regarding outcomes. OBJECTIVE: The objective is to compare characteristics of participants and non participants of two literacy studies. METHODS: Two studies of literacy and health related quality of life were conducted in English- and Spanish-speaking cancer patients. Patients had a range of literacy skills, and each enrolled patient received $20. RESULTS: Nine hundred and twenty-two English-speaking patients were approached. Among the 651 who met eligibility criteria, 420 were enrolled (64.5 per cent). Four hundred and eighty-seven Spanish-speaking patients were approached. Among the 455 who met eligibility criteria, 414 were enrolled (91.0 per cent) (p<0.001). Multiple imputations were performed to impute missing data. Multivariable logistic regression revealed that recruiting site was the only factor predictive of enrollment in Spanish-speaking patients. Age, education, and recruiting site were important predictors in English-speaking patients. Sensitivity analysis using patients with complete data generated similar results. CONCLUSIONS: Spanish-speaking patients enrolled at a much higher rate than English-speaking patients, which is encouraging for future research in this underserved population. One important literacy-related factor (education) did not affect enrollment in Spanish-speaking patients, suggesting that there was no selection bias in this study. Recruiting sites with more indigent patients and long clinic waiting times had higher enrollment, suggesting that monetary compensation and time availability may be important considerations in study participation. PMID- 18381708 TI - Repeated events and total time on test. AB - We adopt the total time on test procedure to investigate monotone time trends in the intensity in a repeated event setting. The correct model is assumed to be a proportional hazards model, with a random effect to account for dependence within subjects. The method offers a simple routine for testing relevant hypotheses for recurrent event processes, without making distributional assumptions about the frailty. Such assumptions may severely affect conclusions concerning regression coefficients and cause bias in the estimated heterogeneity. The method is illustrated by re-analyzing Danish registry data and a long-term Swiss clinical study on recurrence in affective disorder. PMID- 18381707 TI - Estimating selection pressures on HIV-1 using phylogenetic likelihood models. AB - Human immunodeficiency virus (HIV-1) can rapidly evolve due to selection pressures exerted by HIV-specific immune responses, antiviral agents, and to allow the virus to establish infection in different compartments in the body. Statistical models applied to HIV-1 sequence data can help to elucidate the nature of these selection pressures through comparisons of non-synonymous (or amino acid changing) and synonymous (or amino acid preserving) substitution rates. These models also need to take into account the non-independence of sequences due to their shared evolutionary history. We review how we have developed these methods and have applied them to characterize the evolution of HIV-1 in vivo. To illustrate our methods, we present an analysis of compartment specific evolution of HIV-1 env in blood and cerebrospinal fluid and of site-to site variation in the gag gene of subtype C HIV-1. PMID- 18381709 TI - Generalizing from clinical trial data: a case study. The risk of suicidality among pediatric antidepressant users. AB - For the results of randomized controlled clinical trials (RCTs) and related meta analyses to be useful in practice, they must be relevant to a definable group of patients in a particular clinical setting. To the extent this is so, we say that the trial is generalizable or externally valid. Although concern about the generalizability of the results of RCTs is often discussed, there are few examples of methods for assessing the generalizability of clinical trial data. In this paper, we describe and illustrate an approach for making what we call generalizability judgments and illustrate the approach in the context of a case study of the risk of suicidality among pediatric antidepressant users. PMID- 18381710 TI - Bayesian disease mapping using product partition models. AB - Our objective is to develop a model to estimate the relative risk of disease in each area, Ai, i=1, ... , n, of a region and to identify areas of unusually high or low risk. We use a product partition model (PPM) in which we assume that the true relative risks can be partitioned into a number of components or sets of areas where the relative risks are equal. The PPM allows the data to weight those partitions likely to hold and inference about particular parameters may be made by first conditioning on the partition and then averaging over all partitions. We develop Markov chain Monte Carlo (MCMC) techniques to approximate the posterior distributions of the partitions and the parameters. We first test the method in a simulation study and then apply it to data for two separate groups of different types of cancer in the Mid-Western Health Board region in Ireland. The results are compared with those obtained using the standardized mortality ratio method, an empirical Bayes method, a spatial scan method and a nonparametric Bayesian method. PMID- 18381711 TI - Properties of alkali metal atoms deposited on a MgO surface: a systematic experimental and theoretical study. AB - The adsorption of small amounts of alkali metal atoms (Li, Na, K, Rb, and Cs) on the surface of MgO powders and thin films has been studied by means of EPR spectroscopy and DFT calculations. From a comparison of the measured and computed g values and hyperfine coupling constants (hfccs), a tentative assignment of the preferred adsorption sites is proposed. All atoms bind preferentially to surface oxide anions, but the location of these anions differs as a function of the deposition temperature and alkali metal. Lithium forms relatively strong bonds with MgO and can be stabilized at low temperatures on terrace sites. Potassium interacts very weakly with MgO and is stabilized only at specific sites, such as at reverse corners where it can interact simultaneously with three surface oxygen atoms (rubidium and cesium presumably behave in the same way). Sodium forms bonds of intermediate strength and could, in principle, populate more than a single site when deposited at room temperature. In all cases, large deviations of the hfccs from the gas-phase values are observed. These reductions in the hfccs are due to polarization effects and are not connected to ionization of the alkali metal, which would lead to the formation of an adsorbed cation and a trapped electron. In this respect, hydrogen atoms behave completely differently. Under similar conditions, they form (H(+))(e(-)) pairs. The reasons for this different behavior are discussed. PMID- 18381712 TI - Exploring helical folding of oligoureas during chain elongation by high resolution magic-angle-spinning (HRMAS) NMR spectroscopy. AB - The development of novel folding oligomers (foldamers) for biological and biomedical applications requires both precise structural information and appropriate methods to detect folding propensity. However, the synthesis and the systematic conformational investigation of large arrays of oligomers to determine the influence of factors, such as chain length, side chains, and surrounding environment, on secondary structure can be quite tedious. Herein, we show for 2.5 helical N,N'-linked oligoureas (gamma-peptide lineage) that the whole process of foldamer characterization can be accelerated by using high-resolution magic-angle spinning (HRMAS) NMR spectroscopy. This was achieved by monitoring a simple descriptor of conformational homogeneity (e.g., chemical shift difference between diastereotopic main chain CH2 protons) at different stages of oligourea chain growth on a solid support. HRMAS NMR experiments were conducted on two sets of oligoureas, ranging from dimer to hexamer, immobilized on DEUSS, a perdeuterated poly(oxyethylene)-based solid support swollen in solvents of low to high polarity. One evident advantage of the method is that only minute amount of material is required. In addition, the resonance of the deuterated resin is almost negligeable. On-bead NOESY spectra of high quality and with resolution comparable to that of liquid samples were obtained for longer oligomers, thus allowing detailed structural characterization. PMID- 18381713 TI - Theoretical investigation of clusters of phosphorus and arsenic: fascination and temptation of high symmetries. AB - We present a theoretical study of the energetic and thermodynamic stability of selected phosphorus and arsenic clusters containing 18 to 168 atoms. For this purpose we employ MP2 as well as DFT functionals BP86 and B3LYP with extended basis sets. All procedures predict the family of one-dimensional polymers X18+12n, each with 2n-1 isomers of virtually identical energy, to be more stable than other structures investigated so far. Furthermore, islands of stability result for ring-shaped clusters X24n with Dnd symmetry for n=4 (only for arsenic), 5, 6, and 7. Phosphorus and arsenic show otherwise a very similar behavior. An investigation of basis set effects shows that a doubly polarized triple zeta valence basis (TZVPP) is both necessary and sufficient. In comparison to the reliable spin component scaled MP2 (SCS-MP2) procedure, DFT methods underestimate and MP2 overestimates the stability of larger clusters; the discrepancy increases with the number of atoms. The addition of a long-range dispersion correction to B3LYP energies does not rectify the shortcomings of DFT in comparison with SCS-MP2. PMID- 18381714 TI - Encapsulation of ferrocene and peripheral electrostatic attachment of viologens to dimeric molecular capsules formed by an octaacid, deep-cavity cavitand. AB - In aqueous media the deep-cavity cavitand octaacid 1 forms stable dimeric molecular capsules 1(2), which are stabilized by hydrophobic effects. In this work we investigate the binding interactions in aqueous solution between these capsules and the redox active guests, ferrocene (Fc) and three 4,4'-bipyridinium (viologen) dications: methyl viologen (MV(2+)), ethyl viologen (EV(2+)), and butyl viologen (BV(2+)). Using NMR spectroscopic and electrochemical techniques we clearly show that the hydrophobic Fc guest is encapsulated inside 1(2). An interesting effect of this encapsulation is that the reversible voltammetric response of Fc is completely eliminated when it resides inside the 1(2) capsular assembly, a finding that is attributed to very slow electrochemical kinetics for the oxidation of Fc@1(2). Diffusion coefficient measurements (PGSE NMR spectroscopy) reveal that all three viologen guests are strongly bound to the dimeric capsules. However, the (1)H NMR spectroscopic data are not consistent with encapsulation and the measured diffusion coefficients indicate that two viologen guests can strongly associate with a single dimeric capsule. Furthermore, the (V(2+))(2)*1(2) complex is capable of encapsulating ferrocene, clearly suggesting that the viologen guests are bound externally, via coulombic interactions, to the anionic polar ends of the capsule. The electrochemical kinetic rate constants for the reduction of the viologen residue in the V(2+)*1(2) complexes were measured and found to be substantially lower than those for the free viologen guests. PMID- 18381715 TI - Rational modification of ligand-binding preference of avidin by circular permutation and mutagenesis. AB - Chicken avidin is a key component used in a wide variety of biotechnological applications. Here we present a circularly permuted avidin (cpAvd4-->3) that lacks the loop between beta-strands 3 and 4. Importantly, the deletion of the loop has a positive effect on the binding of 4'-hydroxyazobenzene-2-carboxylic acid (HABA) to avidin. To increase the HABA affinity of cpAvd4-->3 even further, we mutated asparagine 118 on the bottom of the ligand-binding pocket to methionine, which simultaneously caused a significant drop in biotin-binding affinity. The X-ray structure of cpAvd4--> 3(N118M) allows an understanding of the effect of mutation to biotin-binding, whereas isothermal titration calorimetry revealed that the relative binding affinity of biotin and HABA had changed by over one billion-fold between wild-type avidin and cpAvd4-->3(N118M). To demonstrate the versatility of the cpAvd4-->3 construct, we have shown that it is possible to link cpAvd4-->3 and cpAvd5-->4 to form the dual-chain avidin called dcAvd2. These novel avidins might serve as a basis for the further development of self-organising nanoscale avidin building blocks. PMID- 18381716 TI - Silica biomineralization in diatoms: the model organism Thalassiosira pseudonana. AB - After complete genome sequencing, the diatom Thalassiosira pseudonana has become an attractive model organism for silica biomineralization studies. Recent progress, especially with respect to intracellular silicic acid processing, as well as to the natures of the biomolecules involved in diatom cell wall formation, is described. On the one hand, considerable progress has been made with respect to silicon uptake by special proteins (SITs) from the surrounding water, as well as to the storage and processing of silicon before cell division. On the other hand, the discovery and characterisation of remarkable biomolecules such as silaffins, polyamines and--quite recently--of silacidins in the siliceous cell walls of diatoms strongly impacts the growing field of biomimetic materials synthesis. PMID- 18381717 TI - Control of porosity geometry in amino acid derived nanoporous materials. AB - Substitution of the pillaring ligand in the homochiral open-framework [Ni(2)(L asp)(2)(bipy)] by extended bipy-type ligands leads to a family of layer structured, homochiral metal-organic frameworks. The 1D channel topology can be modified by the nature of the organic linker, with shape, cross-section and the chemical functionality tuneable. In addition, the volume of these channels can be increased by up to 36 % compared to the parent [Ni(2)(L-asp)(2)(bipy)]. The linker 1,4-dipyridylbenzene (3rbp) gives access to a new layered homochiral framework [Ni(2)(L-asp)(2)(3rbp)] with channels of a different shape. In specific cases, non-porous analogues with the linker also present as a guest can be activated to give porous materials after sublimation. Their CO(2) uptake shows an increase of up to 30 % with respect to the parent [Ni(2)(L-asp)(2)(bipy)] framework. PMID- 18381719 TI - Synthesis and complexation of multiarmed cycloveratrylene-type ligands: observation of the "boat" and "distorted-cup" conformations of a cyclotetraveratrylene derivative. AB - Investigations of a previously reported ligand, hexakis(2 pyridylmethyl)cyclotricatechylene (1), and a new tetrameric bridging ligand, octakis(2-pyridylmethyl)cyclotetracatechylene (2), the latter constructed on a larger cyclotetraveratrylene (CTTV) scaffold, are described. Variable-temperature NMR studies support a "sofa" conformation for 2, akin to studies on the parent compound. The coordination chemistry of 2 and its smaller trimeric homologue have also been investigated with silver(I), copper(II) and palladium(II) salts. An unexpected chelating mode was observed for 1 in the structure of DMF subset[Pd(3)Cl(6)(1)] x DMF, whereby the palladium cations bridge two veratrole subunits rather than chelating within a single subunit. In the structure of [Ag(4)(2)][Co(C(2)B(9)H(11))(2)](4) x 2.8 CH(3)CN x H(2)O, ligand 2 adopts a "boat" conformation, whereas in [Pd(4)Cl(8)(2)] x 4 H(2)O, (1)H NMR spectroscopic studies and calculations indicate that the ligand is present in a previously unobserved "distorted-cup" conformation. This conformation was calculated to be approximately 90 kJ mol(-1) lower in energy than the alternative "sofa" conformation. Thus, coordination-induced conformational control over CTTV derivatives offers new routes to exploit the host-guest chemistry of these compounds. PMID- 18381718 TI - Synthesis, characterization, and remarkable biological properties of cyclodextrins bearing guanidinoalkylamino and aminoalkylamino groups on their primary side. AB - The introduction of aminoalkylamino and guanidinoalkylamino substituents on the primary side of beta- and gamma-cyclodextrin (CDs) resulted in a series of novel compounds that were extensively characterized by NMR spectroscopy and mass spectrometry. Bromination of the primary side of beta- and gamma-CD, and reaction with neat alkylene diamines at a pressure of 7 atm afforded aminoalkylamino derivatives that were then guanylated at the primary amino group to give the corresponding guanidinoalkylamino-CDs. These compounds are water soluble and display pK(a) values that allow them to be mostly protonated at neutral pH; for example, pK(a(1)) approximately 6.4 and pK(a(2)) approximately 9.5 for the aminoethylamino-beta-CD and pK(a(1)) approximately 7.8 and pK(a(2)) approximately 11.0 for the guanidinoethylamino-beta-CD. The title CDs are rigid, cyclic alpha-D glucopyranose oligomers (heptamers or octamers) with branches that resemble lysine and arginine side chains that enable multiple interactions with suitable substrates. Thus, they bear similarities to known cell-penetrating peptides. Indeed, the compounds were found to cross the membranes of HeLa cells and penetrate inside the cytoplasm quickly, the guadinylated ones within 15 min, as shown by fluorescence microscopy using fluorescein-labeled derivatives. The toxicity of the compounds, measured by performing MTT tests, ranged from 50 to 300 microM. Furthermore, some of the aminated CDs could facilitate the transfection of DNA expressing the green fluorescent protein (GFP) in HEK 293T cells, with effectiveness comparable to the commercial agent Lipofectamine 2000. Circular dichroism, atomic force microscopy and electrophoresis experiments confirmed the strong interaction of the compounds with DNA. Because of their carbohydrate, non-peptide nature the title compounds are not anticipated to be enzymatically labile or immunogenic, and thus they fulfill many of the criteria for non-hazardous transport vectors in biological and pharmaceutical applications. PMID- 18381720 TI - Copper(II) coordination chemistry of westiellamide and its imidazole, oxazole, and thiazole analogues. AB - The copper(II) coordination chemistry of westiellamide (H(3)L(wa)), as well as of three synthetic analogues with an [18]azacrown-6 macrocyclic structure but with three imidazole (H(3)L(1)), oxazole (H(3)L(2)), and thiazole (H(3)L(3)) rings instead of oxazoline, is reported. As in the larger patellamide rings, the N(heterocycle)-N(peptide)-N(heterocycle) binding site is highly preorganized for copper(II) coordination. In contrast to earlier reports, the macrocyclic peptides have been found to form stable mono- and dinuclear copper(II) complexes. The coordination of copper(II) has been monitored by high-resolution electrospray mass spectrometry (ESI-MS), spectrophotometric and polarimetric titrations, and EPR and IR spectroscopies, and the structural assignments have been supported by time-dependent studies (UV/Vis/NIR, ESI-MS, and EPR) of the complexation reaction of copper(II) with H(3)L(1). Density functional theory (DFT) calculations have been used to model the structures of the copper(II) complexes on the basis of their spectroscopic data. The copper(II) ion has a distorted square-pyramidal geometry with one or two coordinated solvent molecules (CH(3)OH) in the mononuclear copper(II) cyclic peptide complexes, but the coordination sphere in [Cu(H(2)L(wa))(OHCH(3))](+) differs from those in the synthetic analogues, [Cu(H(2)L)(OHCH(3))(2)](+) (L = L(1), L(2), L(3)). Dinuclear copper(II) complexes ([Cu(II) (2)(HL)(mu-X)](+); X = OCH(3), OH; L = L(1), L(2), L(3), L(wa)) are observed in the mass spectra. While a dipole-dipole coupled EPR spectrum is observed for the dinuclear copper(II) complex of H(3)L(3), the corresponding complexes with H(3)L (L = L(1), L(2), L(wa)) are EPR-silent. This may be explained in terms of strong antiferromagnetic coupling (H(3)L(1)) and/or a low concentration of the dicopper(II) complexes (H(3)L(wa), H(3)L(2)), in agreement with the mass spectrometric observations. PMID- 18381721 TI - Bonding in tropolone, 2-aminotropone, and aminotroponimine: no evidence of resonance-assisted hydrogen-bond effects. AB - The properties of the intramolecular hydrogen bond (IMHB) in tropolone, aminotropone, and aminotroponimine have been compared with those in the corresponding saturated analogues at the B3LYP/6-311+G(3df,2p)//B3LYP/6 311+G(d,p) level of theory. In general, all those compounds in which the seven membered ring is unsaturated exhibit a stronger IMHB than their saturated counterparts. Nevertheless, this enhanced strength is not primarily due to resonance-assisted hydrogen-bond effects, but to the much higher intrinsic basicity and acidity of the hydrogen-bond acceptor and donor groups, respectively, in the unsaturated compounds. These acidity and basicity enhancements have a double origin: 1) the unsaturated nature of the moiety to which the hydrogen-bond donor and acceptor are attached and 2) the cyclic nature of the compounds under scrutiny. As has been found for hydroxymethylene and aminomethylene cyclobutanones, and cyclobutenones and their nitrogen-containing analogues, the IMHB strength follows the [donor, acceptor] trend: [OH, C=NH]>[OH, C=O]>[NH(2), C=NH]>[NH(2), C=O] and fulfills a Steiner-Limbach correlation similar to that followed by intermolecular hydrogen bonds. PMID- 18381722 TI - Nucleophilic additions to alkylidene bis(sulfoxides): stereoelectronic effects in vinyl sulfoxides. AB - Conjugate additions of nucleophiles (e.g. enolates, amines and malonate anions) to bis(p-tolylsulfinyl)alkenes, alkylidene-1,3-dithiane-1,3-dioxides and alkylidene-1,3-dithiolane-1,3-dioxides have recently been published. Reasons for different selectivities and reaction rates will be discussed by consideration of steric and electronic effects. The preferred mode of attack can be explained by stereoelectronic effects (hyperconjugation) in the primarily carbanion, which is stabilized by n-->S-O-sigma* interaction with an antiperiplanar S=O group. Calculation of the transition states [BP86/aug-TZVP] for the addition of acetone enolate to the dithiane-derived alkylidene bis(sulfoxide) revealed that 6.6-7.3 kJ mol(-1) more energy is needed for an attack leading to a less-stabilized carbanion. Two axial S=O groups in dithiolane-derived alkylidene bis(sulfoxides) lead to a higher reactivity towards nucleophiles. PMID- 18381723 TI - Ambident reactivity of the cyanate anion. PMID- 18381724 TI - Cycloisomerization of 1,n-enynes: challenging metal-catalyzed rearrangements and mechanistic insights. AB - Metal-catalyzed cycloisomerization reactions of 1,n-enynes have appeared as conceptually and chemically highly attractive processes as they contribute to the highly demanded search for atom economy and allow the discovery of new reactions. Since the pioneering studies with palladium by the research group of Barry Trost in the mid-1980s, several other metals have been identified as excellent catalysts for the rearrangement of enyne skeletons. Moreover, the behavior of 1,n enynes may be influenced by other functional groups such as alcohols, aldehydes, ethers, alkenes, or alkynes, thus enhancing the molecular complexity of the synthesized products. Apart from the intrinsic rearrangements of 1,n-enynes, several tandem reactions incorporating intramolecular trapping agents or intermolecular partners have been discovered. This Review aims to highlight the main contributions in this field of catalysis and to propose and comment on the mechanistic insights of the recent discoveries. PMID- 18381725 TI - Observation of direct bonds between carbon and nitrogen in Si-B-N-C ceramic after pyrolysis at 1400 degrees C. PMID- 18381726 TI - Total synthesis and structural revision of callipeltoside C. PMID- 18381727 TI - Visual observation of redistribution and dissolution of palladium during the Suzuki-Miyaura reaction. PMID- 18381728 TI - Copper-catalyzed C-C coupling of thiol esters and boronic acids under aerobic conditions. PMID- 18381729 TI - Chiral recognition via helical sense and phase in a crystalline supramolecular array of intermeshed triple-helices. AB - Hexatertiary butyl-substituted D(3)-symmetrical cage ligands composed of octahedral bidentate complexes of Ni(II) or Cu(II) ligated on either side to triskelion arrangements of three salicyl rings bound to a central nitrogen hub atom exhibit triple-helical conformations. Maximization of intermolecular aromatic-aromatic interactions between these complexes promote extended tongue and groove interleaving and affords a supramolecular array of diastereomeric intermeshed C(3)-symmetrical triple-helices perpetuating sideways throughout alternating enantiomeric layer stacks in the entire P-3 space group crystal lattice. Chiral recognition within these supramolecular ensembles is based upon the principles of helical sense and phase. A discussion of the symmetry, mechanical and chemical factors, and constraints influencing the formation of these self-assembled crystalline supramolecular ensembles will be presented. PMID- 18381730 TI - Density functional studies on palladium-catalyzed Suzuki-Miyaura cross-coupling reactions assisted by N- or P-chelating ligands. AB - DFT studies with the B3LYP functional have been carried out on the Suzuki-Miyaura cross-coupling reactions of phenyl chloride and phenylboronic acid catalyzed by palladium complexes with N- or P-chelating ligands. The full catalytic cycle, from the addition of reactants to the catalyst to the release of the cross coupled product from the complexed intermediate, has been examined. The stages within the cycle, such as oxidative addition, transmetalation, and reductive elimination, were validated by linking the mechanistically relevant intermediates and transition states. Various derivatives of diimine, diphosphine, and diamine were considered as potential model ligands. The catalytic reaction employing diimine as the chelating ligand has been verified as the one with the most energetically feasible route. PMID- 18381731 TI - Professor Wolfgang Lindner winner of 2008 Chirality Medal. PMID- 18381732 TI - Analysis of the mechanism of asymmetric amplification by chiral auxiliary trans 1,2-diaminocyclohexane bistriflamide. AB - Asymmetric amplification is a phenomenon in which the enantiomeric excess (ee) of a product is higher than that of a chiral auxiliary for a catalyst. We analyzed the mechanism of asymmetric amplification observed in the addition of diethylzinc (Et(2)Zn) to benzaldehyde (PhCHO) to synthesize 1-phenyl-1-propanol in the presence of trans-1,2-diaminocyclohexane bistriflamide (DCBF) and titanium tetraisopropoxide (TIOP). In a manner similar to the reaction in which 1 piperidino-3,3-dimethyl-2-butanol is a chiral auxiliary for the catalyst, when asymmetric amplification was observed, the ee of the product varied as the reaction progressed. The mechanisms of variation in ee in the two reactions, however, were different. No asymmetric amplification was observed when TIOP and PhCHO were added to a mixture of DCBF and Et(2)Zn, while the ee of the product was always higher than that of DCBF when PhCHO and Et(2)Zn were added to a mixture of DCBF and TIOP. In the latter case, the product ee decreased as the reaction progressed. The results indicate that DCBF forms inactive heterochiral complex causing an increase in the ee of DCBF in the solution, which is the chiral auxiliary for the catalyst. But the complex is not very stable and gradually dissociates due to the reaction with Et(2)Zn. As a result, the asymmetric amplification decreases as the reaction progresses. PMID- 18381733 TI - Highly enantioselective synthesis, crystal structure, and circular dichroism spectroscopy of (R)-bambuterol hydrochloride. AB - The present article describes the asymmetric synthesis of (R)-bambuterol hydrochloride based on 1-(3,5-dihydroxyphenyl)ethanone as starting material, which was esterified by dimethylcarbamic chloride, and brominated by copper (II) bromide. Then the carbonyl group was reduced efficiently using (-)-B chlorodiisopinocamphenylborane [(-)-DIP-chloridetrade mark] as an asymmetrical reducing agent. Followed by epoxide ring closure with NaOH and ring expansion with tert-butylamine led to the desired product (R)-bambuterol with e.e. up to 99%. The optical properties and absolute configuration of (R)-bambuterol hydrochloride were further investigated using circular dichroism spectroscopy and X-ray single crystal analysis. PMID- 18381734 TI - Coordination chemistry of the cyclo-(P(5)tBu(4))(-) ion: monomeric and oligomeric copper(I), silver(I) and gold(I) complexes. AB - [Na{cyclo-(P(5)tBu(4))}] (1) reacts with [CuCl(PCyp(3))(2)] (Cyp=cyclo-C(5)H(9)) and [CuCl(PPh(3))(3)] (1:1) to give the corresponding copper(I) complexes with a tetra-tert-butylcyclopentaphosphanide ligand, [Cu{cyclo- (P(5)tBu(4))}(PCyp(3))(2)] (2) and [Cu{cyclo-(P(5)tBu(4))}(PPh(3))(2)] (3). The CuCl adduct of 2, [Cu(2)(mu-Cl){cyclo-(P(5)tBu(4))}(PCyp(3))(2)] (4), was obtained from the reaction of 1 with [CuCl(PCyp(3))(2)] (1:2). Compounds 2 and 3 rearrange, even at -27 degrees C, to give [Cu(4){cyclo- (P(4)tBu(3))PtBu}(4)] (5), in which ring contraction of the [cyclo-(P(5)tBu(4))](-) anion has occurred. The reaction of 1 with [AgCl(PCyp(3))](4) or [AgCl(PPh(3))(2)] (1:1) leads to the formation of [Ag(4){cyclo-(P(4)tBu(3))PtBu}(4)] (6). Intermediates, which are most probably mononuclear, "[Ag{cyclo-(P(5)tBu(4))}(PR(3))(2)]" (R=Cyp, Ph) could be detected in the reaction mixtures, but not isolated. Finally, the reaction of 1 with [AuCl(PCyp(3))] (1:1) yielded [Au{cyclo-(P(5)tBu(4))}(PCyp(3))] (7), whereas an inseparable mixture of [Au(3){cyclo-(P(5)tBu(4))}(3)] (8) and [Au(4){cyclo-(P(4)tBu(3))PtBu}(4)] (9) was obtained from the analogous reaction with [AuCl(PPh(3))]. Complexes 3-7 were characterised by (31)P NMR spectroscopy, and X-ray crystal structures were determined for 3-9. PMID- 18381736 TI - Energy migration in a self-assembled nonameric porphyrinic molecular box. AB - We describe the construction of self-assembled double-decker porphyrin arrays built up from two covalently connected trimeric Zn-porphyrin units that are joined together by metal-coordination bonds with diamine ligands. We used three different types of diamine ligands: 1,4-diaza[2.2.2]bicyclooctane (DABCO), 4,4' bipyridine (BIPY), and 5,15-bis(4-pyridyl)-10,20-diphenylporphyrin (DPYP). The ligands act as pillars, through two axial coordination bonds with the porphyrinic Zn(II) ions, to block the planes of the porphyrin units in an almost cofacial orientation and inducing the formation of a trigonal prismatic structure. The spectroscopic and photophysical properties of the Zn-trisporphyrin component were determined as well as those of the resulting multimolecular cagelike assemblies. The double-decker assembly with DPYP as the pillars constitutes a nonameric porphyrin aggregate. Although this assembly is thermodynamically less stable than those containing DABCO or BIPY, efficient photoinduced energy transfer occurs (96% yield) from the trisporphyrin base units to the DPYP side walls. The rate of the energy-transfer process is in good agreement with that calculated for a dipole-dipole (Forster) mechanism corrected for the unfavorable orientation geometry of the donor and the axially bound acceptor. PMID- 18381737 TI - Synthesis and application of chiral beta-amino disulfides as ligands for the enantioselective addition of diethylzinc to aldehydes. AB - A new class of chiral beta-amino disulfides was synthesized from readily available and inexpensive starting materials by a straightforward method and their abilities as ligands were examined in the enantioselective addition of diethylzinc to aldehydes. Enantiomeric excesses of up to 99% have been obtained using 0.5 mol % of the chiral catalysts. PMID- 18381735 TI - Synthesis of dendritic stationary phases with surface-bonded L-phenylalanine derivate as chiral selector and their evaluation in HPLC resolution of racemic compounds. AB - Four dendrimers were synthesized on aminopropyl-modified silica gel using methyl acrylate and ethylene diamine as building blocks by divergent method. Four generations of chiral stationary phases (CSPs) were prepared by coupling of L-2 (p-toluenesulfonamido)-3-phenylpropionyl chloride to corresponding dendrimers. The derivatives prepared on silica gel were characterized by FT-IR, (1)H NMR, and elemental analysis. The selector loadings of these four generations of CSPs generally showed a decrease tendency with the increase of generation numbers of dendrimers. The enantioseparation properties of these CSPs were preliminarily investigated by high-performance liquid chromatography. The CSP derived from the three-generation dendrimer exhibited the best enantioseparation capability. Effects of the mobile phase composition and molecular structures of racemic mixtures on enantioseparation were further studied. PMID- 18381738 TI - The phenoxy/phenol/copper cation: a minimalistic model of bonding relations in active centers of mononuclear copper enzymes. AB - The "bare" complex [Cu(PhOH)(PhO)](+) with a phenol (PhOH) and a phenoxy (PhO) ligand bound to copper is studied both experimentally and computationally. The binding energies and structure of this complex are probed by mass spectrometry, infrared multi-photon dissociation, and DFT calculations. Further, the monoligated complexes [Cu(PhO)](+) and [Cu(PhOH)](+) are investigated for comparison. DFT calculations on the [Cu(PhOH)(PhO)](+) complex predict that a phenolate anion interacts with copper(II) preferentially through the oxygen atom, and the bonding is associated with electron transfer to the metal center resulting in location of the unpaired electron at the aromatic moiety. Neutral phenol, on the other hand, interacts with copper preferentially through the aromatic ring. The same arrangements are also found in the monoligated complexes [Cu(PhO)](+) and [Cu(PhOH)](+). The calculations further indicate that the bond strength between the copper atom and the oxygen atom of the phenoxy radical is weakened by the presence of neutral phenol from 2.6 eV in bare [Cu(PhO)](+) to 2.1 eV in [Cu(PhOH)(PhO)](+). PMID- 18381739 TI - Parallel SFC/MS-MUX screening to assess enantiomeric purity. AB - Enantiomeric excess (ee) was evaluated for two internally synthesized compound libraries using a high-throughput automated, intelligent four-channel parallel supercritical fluid chromatography/mass spectrometry system equipped with a multiplexed ion source interface (SFC/MS-MUX). The two libraries contained compounds spanning a wide range of enantiomeric ratios with structurally diverse chemical scaffolds and stereogenic centers. The system analyzed each sample simultaneously against four chiral columns using up to six organic modifiers. Enhancements to our previously published parallel supercritical fluid chromatography/mass spectrometry system were implemented to address the challenges associated with automated trace enantiomer identification and quantitation. A reversal of enantiomer elution order was observed for several samples across multiple CSPs and modifiers. The relationship between elution order and % ee accuracy is presented for compounds exhibiting high, middle and low % ee values. Despite incidences in which the minor enantiomer eluted prior to the major enantiomer with less than baseline resolution, the overall % ee was in agreement with separations in which full baseline resolution was achieved. The methods presented here demonstrate the value and utility of high-throughput ee determinations to support drug discovery and development programs. PMID- 18381740 TI - Alpha-aminophosphonates as novel organocatalysts for asymmetric Michael addition of carbonyl compounds to nitroolefins. AB - A cyclic alpha-aminophosphonate was found to be a novel organocatalyst for Michael type addition reactions of carbonyl compounds to nitroolefins to afford the corresponding adducts in high enantio- and diastereoselectivities. PMID- 18381741 TI - Self-organization of dendritic supermolecules, based on isocyanide-gold(I), copper(I), -palladium(II), and -platinum(II) complexes, into micellar cubic mesophases. AB - First- and second-generation dendrimers with an isocyanide group as the focal functional point (CN-G(n); n: 1,2) and their corresponding organometallic complexes [MCl(CN-G(n))] (M: Au, Cu), [{CuCl(CN-G(n))2}2], and trans-[MI2(CN G(n))2] (M: Pd, Pt) have been synthesized. The free ligands and the first generation complexes do not show mesogenic behavior, but all of the second generation complexes display a thermotropic micellar cubic mesophase, over a large temperature range, and some of them directly at room temperature. The structure of the mesophase consists of the packing of two, discrete polyhedral micellar aggregates in a three-dimensional cubic Im$?bar 3$m lattice. PMID- 18381742 TI - Ring-closing metathesis: novel routes to aromatic heterocycles. AB - Olefin metathesis has been established as an important and general reaction in synthetic organic chemistry. Recently, it has attracted interest as a powerful tool for the construction of aromatic heterocycles. The importance of heteroaromatic motifs in medicinal chemistry and biology, as well as the efficiency and wealth of metathesis transformations, have resulted in significant success in this rapidly developing area. PMID- 18381743 TI - Synthesis of full length PB1-F2 influenza A virus proteins from 'Spanish flu' and 'bird flu'. AB - The proapoptotic influenza A virus PB1-F2 protein contributes to viral pathogenicity and is present in most human and avian isolates. Previous synthetic protocols have been improved to provide a synthetic full length H1N1 type PB1-F2 protein that is encoded by the 'Spanish flu' isolate and an equivalent protein from an avian host that is representative of a highly pathogenic H5N1 'bird flu' isolate, termed SF2 and BF2, respectively. Full length SF2, different mutants of BF2 and a number of fragments of these peptides have been synthesized by either the standard solid-phase peptide synthesis method or by native chemical ligation of unprotected N- and C-terminal peptide fragments. For SF2 chemical ligation made use of the histidine and the cysteine residues located in positions 41 and 42 of the native sequence, respectively, to afford a highly efficient synthesis of SF2 compared to the standard SPPS elongation method. By-product formation at the aspartic acid residue in position 23 was prevented by specific modifications of the SPPS protocol. As the native sequence of BF2 does not contain a cysteine residue two different mutants of BF2 (Y42C) and BF2 (S47C) with appropriate cysteine exchanges were produced. In addition to the full length molecules, fragments of the native sequences were synthesized for comparison of their physical characteristics with those from the H1N1 human isolate A/Puerto Rico/8/34 (H1N1). All peptides were analyzed by mass spectrometry, (1)H NMR spectroscopy, and SDS-PAGE. The protocols allow the synthesis of significant amounts of PB1-F2 and its related peptides. PMID- 18381744 TI - Proteome analysis of non-model plants: a challenging but powerful approach. AB - Biological research has focused in the past on model organisms and most of the functional genomics studies in the field of plant sciences are still performed on model species or species that are characterized to a great extent. However, numerous non-model plants are essential as food, feed, or energy resource. Some features and processes are unique to these plant species or families and cannot be approached via a model plant. The power of all proteomic and transcriptomic methods, that is, high-throughput identification of candidate gene products, tends to be lost in non-model species due to the lack of genomic information or due to the sequence divergence to a related model organism. Nevertheless, a proteomics approach has a great potential to study non-model species. This work reviews non-model plants from a proteomic angle and provides an outline of the problems encountered when initiating the proteome analysis of a non-model organism. The review tackles problems associated with (i) sample preparation, (ii) the analysis and interpretation of a complex data set, (iii) the protein identification via MS, and (iv) data management and integration. We will illustrate the power of 2DE for non-model plants in combination with multivariate data analysis and MS/MS identification and will evaluate possible alternatives. PMID- 18381745 TI - Risk of drug-related problems for various antibiotics in hospital: assessment by use of a novel method. AB - PURPOSE: To investigate the use of antibiotics in hospitals, to explore drug related problems (DRPs) linked to antibiotics and to introduce a novel way of expressing the risks accompanying use of various antibiotics. METHODS: Patients from internal medicine departments in four Norwegian hospitals were prospectively included in 2002. Demographics, drugs used, medical history, laboratory data and clinical/pharmacological risk factors were recorded. DRPs were identified by clinical pharmacists and assessed in multidisciplinary hospital teams. A new term, the drug risk ratio, was established and defined as the number of times the antibiotic was associated with DRPs in relation to the number of times it was used. RESULTS: Out of the 668 patients included, 283 patients (42%) used antibiotics (AB users). AB users were older (76.2 vs. 73.9), used more drugs on admission (5.1 vs. 4.4) and had more DRPs (3.0 vs. 2.2) than non-users. The DRP categories no further need for drug, non-optimal drug and non-optimal dose were most frequently observed. The drug risk ratio, calculated for 12 antibiotic groups, was highest for aminoglycosides (0.77), beta-lactamase-resistant penicillins (0.56), macrolides (0.54) and quinolones (0.48) and lowest for first- and third-generation cephalosporins, 0.17 and 0.13, respectively. CONCLUSIONS: Nearly half of the hospitalised patients were prescribed antibiotics and antibiotic associated DRPs occurred frequently. The drug risk ratio for the different antibiotic groups varied with a factor of six from the lowest to the highest. A high drug risk ratio would alert of antibiotics which require heightened awareness when going to be used in clinical practice. PMID- 18381746 TI - Periostin, secreted from stromal cells, has biphasic effect on cell migration and correlates with the epithelial to mesenchymal transition of human pancreatic cancer cells. AB - Periostin is a secretory protein that has been suggested to function as a cell adhesion molecule and promote the invasiveness or growth rate of tumors. However, little is known about the association of its expression and epithelial to mesenchymal transition (EMT), which is considered to play a crucial role in cancer cell metastasis. Thus, the authors investigated whether periostin could be involved in the process of EMT and the role of this gene in pancreatic cancer development. The expression of periostin was observed mainly in stromal cells but very little in cancer cells by immunohistochemistry and real-time RT-PCR. In vitro, pancreatic stellate cells (PSCs) exhibited a much higher basal expression of periostin compared with cancer cells. Periostin secreted in the supernatant from 293T cells that expressed periostin (approximately 150 ng/ml) inhibited the migration of pancreatic cancer cells. Coculture assay revealed that periostin expression in PSC was induced by pancreatic cancer cells. To assess the direct role of periostin in pancreatic cancer cells, the authors generated pancreatic cancer cell lines that stably express periostin. The induced expression of periostin (to 150 ng/ml) altered the morphology of cancer cells, changing them from mesenchymal to epithelial phenotypes with the induction of epithelial markers and a reduction of mesenchymal markers, and showed reduced cell migration in vitro and formed smaller tumors as well as suppressed metastasis in vivo. On the other hand, high concentration of recombinant periostin (1 microg/ml) promoted cell migration with AKT activation. The findings suggest that periostin has biphasic effect on the development of pancreatic cancer. PMID- 18381747 TI - Selective inhibitors of picornavirus replication. AB - Picornaviruses cover a large family of pathogens that have a major impact on human but also on veterinary health. Although most infections in man subside mildly or asymptomatically, picornaviruses can also be responsible for severe, potentially life-threatening disease. To date, no therapy has been approved for the treatment of picornavirus infections. However, efforts to develop an antiviral that is effective in treating picornavirus-associated diseases are ongoing. In 2007, Schering-Plough, under license of ViroPharma, completed a phase II clinical trial with Pleconaril, a drug that was originally rejected by the FDA after a New Drug Application in 2001. Rupintrivir, a rhinovirus protease inhibitor developed at Pfizer, reached clinical trials but was recently halted from further development. Finally, Biota's HRV drug BTA-798 is scheduled for phase II trials in 2008. Several key steps in the picornaviral replication cycle, involving structural as well as non-structural proteins, have been identified as valuable targets for inhibition. The current review aims to highlight the most important developments during the past decades in the search for antivirals against picornaviruses. PMID- 18381748 TI - Can you keep a secret? Confidentiality in psychotherapy. AB - Confidentiality is the secret-keeping duty that arises from the establishment of the professional relationship psychologists develop with their clients. It is a duty created by the professional relationship, it is set forth in the American Psychological Association's (2002) Ethical Principles and Code of Conduct, and it is codified in many state regulations. However, the difference between confidentiality and legal privilege; how, why, and when it can be violated; and the reasons for so doing are not well understood by many practitioners. While on the surface confidentiality might seem to be an easy concept to apply to professional practice, in fact it is quite complex and filled with exceptions that frequently differ from circumstance to circumstance and from state to state. A lack of respect for and a lack of familiarity with the significance of these exceptions could have dire professional consequences. This article reviews the ethical imperative of confidentiality and then provides examples of legal cases that help to better understand its complexity. Then, we offer strategies designed to help metal health practitioners when they are confronted with questions regarding confidentiality and privilege. PMID- 18381749 TI - Informed consent to psychotherapy: protecting the dignity and respecting the autonomy of patients. AB - Well-implemented informed consent procedures demonstrate psychotherapists' respect for clients' right to self-determination and can initiate meaningful contributions to treatment through enhancing mutual trust, building rapport, and facilitating a sense of ownership. This article details key components of informed consent to psychotherapy by placing them within real-world psychotherapy scenarios. We provide information on client-therapist discussions of the nature and course of therapy, fees and payment policies, the involvement of third parties, confidentiality policies, and new and untested treatments. In addition, this article addresses informed consent procedures for individuals with impaired cognitive capacities and under the Health Insurance Portability and Accountability Act of 1996 (HIPAA) regulations. PMID- 18381750 TI - Discrimination of chiral guests by chiral channels: variable temperature studies by SXRD and solid state 13C NMR of the deoxycholic acid complexes of camphorquinone and endo-3-bromocamphor. AB - 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid (deoxycholic acid DCA) is able to discriminate between the R- and S-enantiomers of camphorquinone and endo-(+)-3 bromocamphor and select only the S-enantiomers from a racemic mixture. DCA forms novel well ordered 1:1 adducts with (1S)-(+)-camphorquinone and (1S)-endo-(-)-3 bromocamphor, both of which have been characterized by single crystal X-ray diffraction (SXRD). When DCA is cocrystallized with (RS)-camphorquinone and (RS) endo-3-bromocamphor, 1:1 adducts of the S-enantiomers are produced together with crystals of the free racemic guest. In contrast, in the absence of (1S)-(+) camphorquinone, DCA forms a 2:1 adduct with (1R)-(-)-camphorquinone. In this 2:1 adduct the guest is disordered at ambient temperature and undergoes a phase change in the region 160-130 K similar to that observed for the ferrocene adduct, but with only partial ordering of the guest. The SXRD structure of the low temperature form and the variable temperature (13)C CP/MAS NMR are reported. Cocrystallizing DCA with (1R)-endo-(+)-3-bromocamphor gives the free guest and a glassy solid. PMID- 18381751 TI - S-allylcysteine ameliorates ischemia and reperfusion induced renal damage. AB - The effect of the garlic-derived antioxidant S-allylcysteine (SAC) on renal injury and oxidative stress induced by ischemia and reperfusion (IR) was studied in this work. Rats were anesthetized and subjected to right nephrectomy; 15 min later ischemia was induced for a period of 40 min and then the rats were subjected to a reperfusion period of 6 h after which they were killed to obtain blood and the left kidney. SAC was given at a dose of 100 mg/kg 30 min before nephrectomy, 15 min before ischemia, immediately before reperfusion and 2 h after reperfusion. IR-induced renal injury was evident by the increase in blood urea nitrogen (BUN) and serum creatinine as well as by the renal structural damage which was assessed by histological analysis. IR-induced oxidative stress was evident by the increase in immunostaining with 4-hydroxy-2-nonenal (4-HNE). SAC treatment was able to ameliorate the increase in BUN and serum creatinine and to decrease the structural damage. This protective effect was associated with a decrease in the immunostaining for 4-HNE. It is concluded that the antioxidant properties of SAC are involved in its protective effect on renal ischemia and reperfusion injury. PMID- 18381755 TI - Abstracts of the Society of Hospital Medicine annual meeting, April 3-5, 2008, San Diego, California, USA. PMID- 18381756 TI - Intensive induction chemotherapy followed by high dose chemotherapy with autologous hematopoietic progenitor cell rescue in young children newly diagnosed with central nervous system atypical teratoid rhabdoid tumors. AB - BACKGROUND: Central nervous system (CNS) atypical teratoid rhabdoid tumors (AT/RT) are rare tumors of childhood with a dismal prognosis. Historically, surgery and standard dose chemotherapy have resulted in a median survival of 8.5 months from diagnosis. METHODS: Thirteen children newly diagnosed with CNS AT/RT were treated with either the "Head Start I" (HS I) or "Head Start II" (HS II) regimens. Therapy included resection followed by five cycles of cisplatin, vincristine, cyclophosphamide, and etoposide. High dose methotrexate was added to each of the five induction courses in HS II. Consolidation for both regimens included carboplatin, thiotepa, and etoposide with autologous hematopoietic progenitor cell rescue (AHPCR). RESULTS: Six children, median age of 36 months, were treated on HS I between 1992 and 1997. Seven children, median age of 28 months, were treated on HS II between 1997 and 2002. One patient received craniospinal irradiation following AHPCR but prior to recurrence. There are presently three event-free survivors 42+, 54+, and 67+ months following diagnosis; none received RT. All three survivors were enrolled on HS II. Eight patients died of disease (six on HS I); one patient died from infection; one patient died from secondary malignancy following treatment for recurrent AT/RT. CONCLUSION: Three of seven children with CNS AT/RT treated on HS II have experienced long term remissions. Long term survival can be achieved in a subset of young children with CNS AT/RT following resection with the use of multi-drug chemotherapy including high dose methotrexate and myeloablative chemotherapy without radiation therapy (RT). PMID- 18381757 TI - Pro-tolerogenic effects of photodynamic therapy with TH9402 on dendritic cells. AB - A major goal in transplantation immunology is to develop strategies that can specifically promote tolerance to foreign cells and tissues without compromising other immune functions. Experimental induction of transplantation tolerance by dendritic cells (DCs) and/or T regulatory (Treg) cells can efficiently prevent graft-versus-host disease and organ graft rejection in animal models, and there is much hope that similar strategies in transplanted patients will provide an alternative to immunosuppression. Photodynamic therapy (PDT) is a therapeutic treatment for graft versus host disease and organ rejection that may operate via induction of tolerance. We investigated whether a new PDT therapy, based on exposure of cells to 4,5-dibromorhodamine methyl ester (TH9402), may operate via induction of tolerogenic DC. We developed an in vitro model to mimic the in vivo effect of re-infusing peripheral blood mononuclear cells (PBMCs) treated with PDT based on TH9402 (TH-PDT). TH-PDT-treated PBMCs were co-cultured with allogeneic immature monocyte-derived DCs. After 24 h, the phenotype and T-cell stimulatory capacity of the DCs was assessed. Following phagocytosis of TH-PDT PBMCs, DCs maintained an immature phenotype, produced significantly increased amounts of interleukin-10, and had a reduced allostimulatory capacity in comparison to mature DCs. In the context of transplantation, these data suggest that repeated exposure of circulating DCs to TH-PDT PBMCs may result in presentation of alloantigens under anti-inflammatory conditions and induction of antigen-specific tolerance. PMID- 18381758 TI - A cytoprotective role for the heme oxygenase-1/CO pathway during neural differentiation of human mesenchymal stem cells. AB - The inducible protein heme oxygenase-1 (HO-1) catalyzes the oxidation of heme to carbon monoxide (CO) and biliverdin, which play a concerted action in cytoprotection against oxidative stress and in the modulation of cell proliferation and differentiation. Here we report that both HO-1 expression and activity can be highly increased in undifferentiated human mesenchymal stem cells (MSCs) treated with hemin, a known HO-1 inducer. However, HO-1 mRNA and protein expression gradually decrease when MSCs undergo neural differentiation in vitro, making them extremely susceptible to glutamate-mediated cytotoxicity. A time course for HO-1 revealed that this protein is markedly down-regulated after 2 days and returns to control levels 6 days after differentiation. Treatment with glutamate (250 microM) after 2 days of neural differentiation resulted in a more pronounced lactate dehydrogenase release, a marker of cell injury, compared with undifferentiated cells. Notably, cells pretreated with hemin (50 microM) or compounds that release small amounts of CO (10 microM CORM-3 and CORM-A1) rendered cells more resistant to glutamate-induced toxicity; this effect was evident in both undifferentiated and differentiated MSCs. Our findings indicate that MSCs become more vulnerable to oxidative injury during the early stages of differentiation via mechanisms that involve a temporary inhibition of HO-1 expression. Thus, overexpression of HO-1 and CO-releasing molecules could provide a possible therapeutic strategy to improve cell viability during neural differentiation in applications that use stem cell technology. PMID- 18381760 TI - Distribution of mitochondria along small-diameter myelinated central nervous system axons. AB - Small-diameter myelinated CNS axons are preferentially affected in multiple sclerosis (MS) and in the hereditary spastic paraplegias (HSP), in which the distal axon degenerates. Mitochondrial dysfunction has been implicated in the pathogenesis of these and other disorders involving axonal degeneration. The aim of this study was to determine whether the frequency of axonal mitochondria changes along the length of small-diameter fibers and whether there is a preferential localization to the region of the node of Ranvier. We find that mitochondrial numbers do not change along the length of a myelinated small diameter fiber, and, in contrast to the peripheral nervous system, there is no tendency for mitochondrial numbers to increase at the node. PMID- 18381759 TI - The spleen contributes to stroke-induced neurodegeneration. AB - Stroke, a cerebrovascular injury, is the leading cause of disability and third leading cause of death in the world. Recent reports indicate that inhibiting the inflammatory response to stroke enhances neurosurvival and limits expansion of the infarction. The immune response that is initiated in the spleen has been linked to the systemic inflammatory response to stroke, contributing to neurodegeneration. Here we show that removal of the spleen significantly reduces neurodegeneration after ischemic insult. Rats splenectomized 2 weeks before permanent middle cerebral artery occlusion had a >80% decrease in infarction volume in the brain compared with those rats that were subjected to the stroke surgery alone. Splenectomy also resulted in decreased numbers of activated microglia, macrophages, and neutrophils present in the brain tissue. Our results demonstrate that the peripheral immune response as mediated by the spleen is a major contributor to the inflammation that enhances neurodegeneration after stroke. PMID- 18381761 TI - High doses of nicotinamide prevent oxidative mitochondrial dysfunction in a cellular model and improve motor deficit in a Drosophila model of Parkinson's disease. AB - Nicotinamide, the principal form of niacin (vitamin B3), has been proposed to be neuroprotective in Parkinson's disease. However, the effects and mechanisms of nicotinamide on motor function in animals and on mitochondrial function in cellular systems have not been well studied. We hypothesized that niacin-derived NAD(P)H as antioxidants and enzyme cofactors could inhibit oxidative damage and improve mitochondrial function and thus protect neurodegeneration and improve motor function. In the present study, the effects of nicotinamide on mitochondrial function and oxidative stress were studied in a 1-methyl-4 phenylpyridinium (MPP(+))-induced cellular model of Parkinson's disease, and the effects of improving motor dysfunction were studied in an alpha-synuclein transgenic Drosophila Parkinson's model. Mitochondrial function was tested by measuring the activity of mitochondrial complex I and alpha-ketoglutarate dehydrogenase, and oxidative damage was tested by measuring reactive oxygen species, DNA damage (8-oxo-7,8-dihydro-2'-deoxyguanosine and Comet assay), and protein oxidation (protein carbonyls) levels. Nicotinamide at a relatively higher concentration, that is, 100-fold of the level in the cell culture medium (101 mg/L), significantly protected SK-N-MC human neuroblastoma cells from an MPP(+) induced decrease in cell viability, complex I and alpha-ketoglutarate dehydrogenase activity, and an increase in oxidant generation, DNA damage, and protein oxidation. In the Drosophila model, nicotinamide at 15 and 30 mg/100 g diet significantly improved climbing ability. These results suggest that nutritional supplementation of nicotinamide at high doses decreases oxidative stress and improves mitochondrial and motor function in cellular and/or Drosophila models and may be an effective strategy for preventing and ameliorating Parkinson's disease. PMID- 18381762 TI - Neuronal coupling via connexin36 contributes to spontaneous synaptic currents of striatal medium-sized spiny neurons. AB - Gap junctions provide a means for electrotonic coupling between neurons, allowing for the generation of synchronous activity, an important contributor to learning and memory. Connexin36 (Cx36) is largely neuron specific and provides a target for genetic manipulation to determine the physiological relevance of neuronal coupling. Within the striatum, Cx36 is more specifically localized to the interneuronal population, which provides the main inhibitory input to the principal projection medium-sized spiny neurons. In the present study, we examined the impact of genetic ablation of Cx36 on striatal spontaneous synaptic activity. Patch-clamp recordings were performed from medium-sized spiny neurons, the primary target of interneurons. In Cx36 knockout mice, the frequencies of both excitatory and inhibitory spontaneous postsynaptic currents were reduced. We also showed that activation of dopamine receptors differentially modulated the frequency of GABAergic currents in Cx36 knockout mice compared with their wild type littermates, suggesting that dopamine plays a role in altering the coupling of interneurons. Taken together, the present findings demonstrate that electrical coupling of neuronal populations is important for the maintenance of normal chemical synaptic interactions within the striatum. PMID- 18381763 TI - Role for nitric oxide in permeability of hippocampal neuronal hemichannels during oxygen glucose deprivation. AB - Increased hemichannel opening induced by oxygen glucose deprivation (OGD) was reported in the hippocampal pyramidal neuron. It was suggested that the pannexin1 hemichannel opening could mediate ionic flux dysregulation, anoxic depolarization, and energy-depleting efflux of glucose and ATP for ischemic neurons. However, the regulatory mechanisms of pannexin1 hemichannel opening have been poorly understood. Here we showed that excessive generation of nitric oxide (NO) during ischemia could induce the calcein leakage from neurons, which was markedly reduced by NO synthase inhibitor. The calcein leakage from neurons during OGD was also attenuated by the application of N-ethylmaleimide (NEM), an SH-alkylating agent, and dithiothreitol (DTT), a reducer of oxidized sulfhydryl groups. However, the soluble guanylyl cyclase (sGC) inhibitor had a minor effect on the calcein leakage during OGD. Furthermore, the elevated intracellular but not extracellular levels of glutathione could also inhibit the calcein leakage during OGD. Similar results were observed in metabolic inhibition (MI), which is another ischemic-like condition. Finally, immunocytochemical and immunoblotting analysis revealed that, after 1 hr of OGD stimulation, the distribution and expression of pannexin1 showed no significant difference compared with control. However, the pannexin1 mRNA expression was elevated after 1 hr of OGD and a sustained increase was maintained during reperfusion. These results implied that the reactive oxygen species (ROS), especially NO, might be involved in the enhanced pannexin1 hemichannel opening and that the S-nitrosylation but not the NO/cGMP pathway played a more important role in this event. PMID- 18381765 TI - Surface protein patterns govern morphology, proliferation, and expression of cellular markers but have no effect on physiological properties of cortical precursor cells. AB - The ability to differentiate and give rise to neurons, astrocytes, and oligodendrocytes is an inherent feature of neural stem cells, which raises hopes for cell-based therapies of neurodegenerative diseases. However, there are many hurdles to cross before such regimens can be applied clinically. A considerable challenge is to elucidate the factors that contribute to neural differentiation. In this study, we evaluated the possibility of steering neuronal maturation by growing cortical precursor cells on microscale surface patterns of extracellular matrix (ECM) proteins. When the cells were encouraged to extend processes along lines of ECM proteins, they displayed a much more mature morphology, less proliferation capacity, and greater expression of a neuronal marker in comparison with cells grown in clusters on ECM dots. This implied that the growth pattern alone could play a crucial role for neural differentiation. However, in spite of the strikingly different morphology, when performing whole-cell patch-clamp experiments, we never observed any differences in the functional properties between cells grown on the two patterns. These results clearly demonstrate that morphological appearances are not representative measures of the functional phenotype or grade of neuronal maturation, stressing the importance of complementary electrophysiological evidence. To develop successful transplantation therapies, increased cell survival is critical. Because process bearing neurons are sensitive and break easily, it would be of clinical interest to explore further the differentiating capacity of the cells cultured on the ECM dot pattern, described in this article, which are devoid of processes but display the same functional properties as neurons with mature morphology. PMID- 18381764 TI - Selective death of newborn neurons in hippocampal dentate gyrus following moderate experimental traumatic brain injury. AB - Memory impairment is one of the most significant residual deficits following traumatic brain injury (TBI) and is among the most frequent complaints heard from patients and their relatives. It has been reported that the hippocampus is particularly vulnerable to TBI, which results in hippocampus-dependent cognitive impairment. There are different regions in the hippocampus, and each region is composed of different cell types, which might respond differently to TBI. However, regional and cell type-specific neuronal death following TBI is not well described. Here, we examined the distribution of degenerating neurons in the hippocampus of the mouse brain following controlled cortical impact (CCI) and found that the majority of degenerating neurons observed were in the dentate gyrus after moderate (0.5 mm cortical deformation) CCI-TBI. In contrast, there were only a few degenerating neurons observed in the hilus, and we did not observe any degenerating neurons in the CA3 or CA1 regions. Among those degenerating cells in the dentate gyrus, about 80% of them were found in the inner granular neuron layer. Analysis with cell type-specific markers showed that most of the degenerating neurons in the inner granular neuron layer are newborn immature neurons. Further quantitative analysis shows that the number of newborn immature neurons in the dentate gyrus is dramatically decreased in the ipsilateral hemisphere compared with the contralateral side. Collectively, our data demonstrate the selective death of newborn immature neurons in the hippocampal dentate gyrus following moderate injury with CCI in mice. This selective vulnerability of newborn immature dentate neurons may contribute to the persistent impairment of learning and memory post-TBI and provide an innovative target for neuroprotective treatment strategies. PMID- 18381766 TI - Lymphoma presenting as an echogenic periportal mass: sonographic findings. AB - Abdominal lymphoma usually appears as homogeneous, markedly hypoechoic masses on sonography. We report a case of primary lymphoma presenting as an extrahepatic and intrahepatic periportal echogenic mass encasing the hepatic vessels. PMID- 18381768 TI - Quantitative multifocal fMRI shows active suppression in human V1. AB - Multifocal functional magnetic resonance imaging has recently been introduced as an alternative method for retinotopic mapping, and it enables effective functional localization of multiple regions-of-interest in the visual cortex. In this study we characterized interactions in V1 with spatially and temporally identical stimuli presented alone, or as a part of a nine-region multifocal stimulus. We compared stimuli at different contrasts, collinear and orthogonal orientations and spatial frequencies one octave apart. Results show clear attenuation of BOLD signal from the central region in the multifocal condition. The observed modulation in BOLD signal could be produced either by neural suppression resulting from stimulation of adjacent regions of visual field, or alternatively by hemodynamic saturation or stealing effects in V1. However, we find that attenuation of the central response persists through a range of contrasts, and that its strength varies with relative orientation and spatial frequency of the central and surrounding stimulus regions, indicating active suppression mechanisms of neural origin. Our results also demonstrate that the extent of the signal spreading is commensurate with the extent of the horizontal connections in primate V1. PMID- 18381767 TI - Assimilation and accommodation patterns in ventral occipitotemporal cortex in learning a second writing system. AB - Using fMRI, we compared the patterns of fusiform activity produced by viewing English and Chinese for readers who were either English speakers learning Chinese or Chinese-English bilinguals. The pattern of fusiform activity depended on both the writing system and the reader's native language. Native Chinese speakers fluent in English recruited bilateral fusiform areas when viewing both Chinese and English. English speakers learning Chinese, however, used heavily left lateralized fusiform regions when viewing English, but recruited an additional right fusiform region for viewing Chinese. Thus, English learners of Chinese show an accommodation pattern, in which the reading network accommodates the new writing system by adding neural resources that support its specific graphic requirements. Chinese speakers show an assimilation pattern, in which the reading network established for L1 includes procedures sufficient for the graphic demands of L2 without major change. PMID- 18381769 TI - Cortical responses to self and others. AB - The extrastriate body area (EBA) is one among the multiple, functionally specialized regions of the human visual cortex exhibiting modulation by body related stimuli. Here we investigate whether activation patterns differ for the perception of one's own body and the bodies of others. We used functional magnetic resonance imaging to identify body-related brain areas and to see how these areas differentiate between images of one's own body and those of others in the absence of facial or motion cues. Whole brain explorative group-level analysis identified body-related blood oxygen level dependent (BOLD) signal enhancement in five regions of the right and in one region of the left hemisphere (right: in the extrastriate visual and parietal cortex and in the precentral gyrus, left: in the extrastriate visual cortex). General linear model group-level random effects analysis of the self-other contrast revealed self-related responses in the extrastriate and parietal regions in the right hemisphere but also in the right middle frontal gyrus. These results suggest the existence of a cortical network for the extraction of body-related information and another cortical network for the extraction of self-related body information. The two networks partially overlap in the right superior and inferior parietal cortices, but are clearly segregated in the extrastriate visual cortex and in the middle frontal gyrus. In addition, we report that the classical EBA is only involved in the analysis of body-related information but not in the assignment of body identity. The latter appears to be accomplished by a network in right hemisphere comprising the fusiform body area, regions of the superior parietal lobe, the inferior parietal cortex, and the middle frontal gyrus. PMID- 18381770 TI - Social cognition and the brain: a meta-analysis. AB - This meta-analysis explores the location and function of brain areas involved in social cognition, or the capacity to understand people's behavioral intentions, social beliefs, and personality traits. On the basis of over 200 fMRI studies, it tests alternative theoretical proposals that attempt to explain how several brain areas process information relevant for social cognition. The results suggest that inferring temporary states such as goals, intentions, and desires of other people even when they are false and unjust from our own perspective--strongly engages the temporo-parietal junction (TPJ). Inferring more enduring dispositions of others and the self, or interpersonal norms and scripts, engages the medial prefrontal cortex (mPFC), although temporal states can also activate the mPFC. Other candidate tasks reflecting general-purpose brain processes that may potentially subserve social cognition are briefly reviewed, such as sequence learning, causality detection, emotion processing, and executive functioning (action monitoring, attention, dual task monitoring, episodic memory retrieval), but none of them overlaps uniquely with the regions activated during social cognition. Hence, it appears that social cognition particularly engages the TPJ and mPFC regions. The available evidence is consistent with the role of a TPJ related mirror system for inferring temporary goals and intentions at a relatively perceptual level of representation, and the mPFC as a module that integrates social information across time and allows reflection and representation of traits and norms, and presumably also of intentionality, at a more abstract cognitive level. PMID- 18381771 TI - An entorhinal cortex sulcal pattern is associated with Alzheimer's disease. AB - OBJECTIVES: Magnetic resonance (MRI) studies rely on sulcal boundaries to delineate the human entorhinal cortex (EC) and typically show that EC size is reduced in Alzheimer's disease (AD) and a predictor of future dementia. However, it is unknown if variations in the EC sulcal patterns are associated with AD. We classified the lateral EC sulcal boundary as either a rhinal or collateral pattern and tested the hypotheses that the rhinal pattern was (1) more common in AD and (2) associated with a smaller EC size. EXPERIMENTAL DESIGN: MRI was used to determine the prevalence of the rhinal and collateral EC patterns in 421 subjects (212 AD, 107 old normal (ONL), and 102 young NL (YNL). Anatomical validation studies of normal subjects were conducted at postmortem in 34 brain hemispheres and in vivo with 21 MRI volume studies. EC pattern reliability was studied with MRI in both cross-sectional and longitudinal designs. PRINCIPAL OBSERVATIONS: The rhinal pattern was more frequent in the right hemisphere in AD (47%) compared with ONL (28%, odds ratio = 2.25, P = 0.001). EC pattern was not related to ApoE genotype. The validations showed that the EC sulcal pattern was not associated with the neuronal number, surface area, or volume of the EC. In patients with antemortem MRI studied at postmortem it was equivalently determined, that EC patterns are reliably determined on MRI and do not change with the progressive atrophy of AD. CONCLUSIONS: The data indicate that the right hemisphere rhinal pattern is over represented in AD as compared with control. However, in normal subjects the EC rhinal pattern is not associated with a diminished EC tissue size. It remains to be demonstrated if the right EC rhinal sulcus pattern association with AD reflects genetic or developmental influences. PMID- 18381773 TI - Prenatal ultrasound detection of facial clefts: a prospective study of 49,314 deliveries in a non-selected population in Norway. AB - OBJECTIVES: To evaluate prenatal detection of facial clefts by ultrasound examination in a large non-selected population, and to study trends in detection rates over 18 years, as well as the prevalence of isolated cases and those with associated anomalies. METHODS: This prospective follow-up study from January 1987 to December 2004 was divided into two 9-year periods. The study included all registered cases of prenatally or postnatally diagnosed facial clefts in a non selected population in Norway. RESULTS: A total of 101 fetuses or newborns with facial clefts in a population of 49 314 deliveries were registered. The distribution of clefts was: 25 (25%) cleft lip, 52 (51%) cleft lip and palate, and 24 (24%) cleft palate (CP). No CP was detected prenatally. Cleft lip with or without cleft palate (CL(P)) was detected prenatally in 35/77 (45%) cases, with a significant increase in the detection rate from 34% to 58% between the two 9-year periods (P = 0.03). Over the whole study period CL(P) was detected at a median of 19 + 2 gestational weeks, with no change over time; altogether 24/35 (69%) cases were detected at the routine second-trimester ultrasound examination. Thirty three of 77 (43%) cases of CL(P) and 14/24 (58%) cases of CP had associated anomalies; 12/101 (12%) had chromosomal aberrations. In 18/101 (18%) the clefts were part of a syndrome or sequence. CONCLUSIONS: The detection rate for CL(P) improved significantly over time. Detection of CL(P) is important because nearly half the cases have associated anomalies. PMID- 18381774 TI - Lessons from TGN1412 and TARGET: implications for observational studies and meta analysis. AB - Two very different studies are examined: the first, a very large trial in osteoarthritis (the so-called TARGET study) and the second a very small 'first-in man' study of the monoclonal antibody TGN1412. In each trial the unbiased estimate of the treatment effect is not efficient and in con sequence the efficient estimate is not unbiased. In the case of the large trial it seems reasonable that unbiased estimation is desirable but in the case of the small trial it leads to absurd conclusions. These two cases are examined in detail and some general lessons for the analysis of clinical trials and observational studies and collections of studies are drawn. PMID- 18381772 TI - Adaptive evolution of mammalian aromatases: lessons from Suiformes. AB - Estrogen synthesis evolved in chordates to control reproduction. The terminal enzyme in the cascade directly responsible for estrogen synthesis is aromatase cytochrome P450 (P450arom) encoded by the CYP19 gene. Mammals typically have a single CYP19 gene but pigs, peccaries and other Suiformes have two or more resulting from duplication in a common ancestor. Duplication of CYP genes in the steroid synthetic cascade has occurred for only one other enzyme, also terminal, 11beta-hydroxylase P450 (P450c11). P450arom and P450c11 share common substrates and even physiological functions as possible remnants from a common P450 progenitor, perhaps an ancestral P450arom, which is supported by phylogenetic analysis. Conserved tissue-specific expression patterns of P450arom paralogs in placenta and gonads of pigs and peccaries suggest how functional adaptation may have proceeded divergently and influenced adopted reproductive strategies including ovulation rate and litter size. Data suggest that the porcine placental paralog evolved catalytically to protect female conceptuses from testosterone produced by male siblings; the gonadal paralog to synthesize a novel, nonaromatizable testosterone metabolite (1OH-testosterone) that may increase ovulation rate. This would represent a coevolution facilitating litter bearing as pigs diverged from peccaries. Evidence of convergence between the peccary CYP19 genes and lower tissue expression may therefore represent initiation of loss of the functional paralogs. Studies on the Suiforme aromatases provide insights into the evolution of the steroidogenic cascade and metabolic pathways in general, how it translates into physiological adaptations (altered reproductive strategies for instance), and how duplicated genes become stabilized or disappear from genomes. PMID- 18381775 TI - Evaluation of the environmental fate of thymol and phenethyl propionate in the laboratory. AB - BACKGROUND: The natural monoterpenoid pesticides thymol and phenethyl propionate (PEP) are used indoors and outdoors, but their fate in the environment has not been reported. In order better to understand their impact on the environment, water metabolism and soil metabolism studies were conducted with thymol and PEP at a concentration of 10 microg g(-1) in water and in soil under laboratory conditions. RESULTS: Dissipation half-lives of thymol and PEP were 16 and 5 days in water and 5 and 4 days in soil. 2-Phenylethanol and 2-(4-hydroxyphenyl)ethanol were detected as primary degradation products of PEP. Over time, a considerable volatilization loss of thymol, but not of phenethyl propionate, was found in the 1 month study under the experimental conditions used. Less than 6% of thymol and PEP were detected as bound residues, and less than 3% were mineralized during the 30 day study. CONCLUSION: In order to maximize the pesticidal effect, more attention should be paid to the temperature for thymol than for PEP when they are being applied, owing to the high volatility of the former. Thymol and PEP pose low risks to the ecosystem because of their rapid dissipation and low bound residues in the environment. PMID- 18381776 TI - Dissection of the ascending aorta in a patient with HLA-B27 associated ankylosing spondylitis. AB - We describe a 38-year-old patient with ankylosing spondylitis complicated by a non-traumatic dissection of the ascending aorta without concomitant Marfan's syndrome. PMID- 18381777 TI - Childhood chronic recurrent multifocal osteomyelitis: pamidronate therapy decreases pain and improves vertebral shape. AB - OBJECTIVE: Chronic relapsing multifocal osteomyelitis (CRMO) results in significant morbidity, especially in those with vertebral collapse. Symptomatic benefit with intravenous pamidronate (PAM) has been shown; however, few studies have demonstrated radiological benefit. We describe clinical and radiological data on 7 pediatric cases of CRMO treated with PAM. METHODS: Retrospective chart review on all children with CRMO treated with PAM. Response to PAM was measured by subjective reports and radiology including vertebral morphometry. RESULTS: Seven patients (1 male) presented with bone pain at a median age of 8 years (range 5-14). Symptoms had been present for a median of 18 months (range 11-51) before PAM therapy. All patients had involvement of multiple nonspinal sites, 5 children had spinal involvement with vertebral fractures, and 5 had joint involvement. Six cases had symptomatic improvement within 6 months of starting PAM, which was sustained during PAM therapy (median 26 mo, range 6-41) and persisted in the 4 cases who had ceased treatment for the duration of followup (27 mo, range 18-51). The least benefit was seen in the 3 cases with synovial joint involvement. The 3 cases with spinal radiological followup showed modeling of vertebral fractures and in one patient improvement in kyphosis. No radiological improvement in nonspinal lesions was seen. CONCLUSION: PAM therapy was associated with symptomatic improvement and vertebral modeling in children with CRMO. We suggest that children with bone pain and/or spinal involvement be considered for PAM therapy early after diagnosis. PMID- 18381778 TI - Takayasu arteritis in children. AB - OBJECTIVE: To retrospectively evaluate the clinical features, angiographic findings, and outcomes of children with Takayasu arteritis (TA) in Turkey. METHODS: Clinical, laboratory, and angiographic findings and outcomes of 19 children with TA were evaluated with a retrospective chart review. The criteria for inclusion were those proposed by the American College of Rheumatology. RESULTS: Mean followup period was 35.89 +/- 40.75 months (range 1-168, median 30). There were 14 girls and 5 boys. The mean age at diagnosis was 12.84 +/- 2.69 years (range 8-17, median 13). The most common complaints on admission were headache (84%), abdominal pain (37%), claudication of extremities (32%), fever (26%), and weight loss (10%). One patient presented with visual loss. Examination on admission revealed hypertension (89%), absent pulses (58%), and bruits (42%). Angiography revealed type I in 13 patients (aortic arch, descending thoracic, and abdominal aorta), type II in 4 (descending thoracic aorta and abdominal aorta), and type IV in 2 (diffuse aortic and pulmonary artery). The most commonly involved vessels were the renal, subclavian, and carotid arteries. All patients received corticosteroid therapy, and further immunosuppressive therapy was added in 15 patients. Fourteen of the 17 hypertensive patients had renal artery stenosis and 9 underwent surgery or interventional therapy. Thoraco-abdominal bypass graft was performed in 2 patients who had abdominal aortic stenosis. CONCLUSION: Hypertension is the most common clinical feature at presentation. Corticosteroid and immunosuppressive therapy was effective in the control of disease activity. Angioplasty or bypass grafting was successfully performed when needed. PMID- 18381779 TI - Longterm outcome of amyloidosis associated with juvenile idiopathic arthritis. AB - OBJECTIVE: To determine the outcome of amyloidosis associated with juvenile idiopathic arthritis (JIA) in a hospital-based series. METHODS: Patient registers and amyloidosis biopsy files of the Department of Pediatrics of Rheumatism Foundation Hospital, the main tertiary center for inflammatory joint disorders in children in Finland, were scrutinized from 1976 to the end of 2003 to look for amyloidosis in patients under age 19 years. Medical records were reviewed and patients were interviewed by telephone. The causes of any deaths were obtained from death certificates. RESULTS: Twenty-four patients under age 19 years with biopsy-proven amyloidosis were found. As a sign of renal disease at the time of diagnosis of amyloidosis, 16 patients (67%) had proteinuria, but none had renal insufficiency. The 5-year survival rate of the series was 87.5% (95% CI 75% to 100%), and 10-year survival was 75% (54% to 92%). Ten patients (42%) out of the 24 died during a mean followup of 15.4 (range 1.5-27.6) years. The main cause of death was related to JIA in all patients but one. Patients treated with prednisolone alone from the diagnosis of amyloidosis onward had a mortality rate significantly higher than those taking disease modifying antirheumatic drugs and/or cytostatics (p = 0.001). At the end of followup, 14 patients (58%) were alive, 12 with normal renal function (3 of them had undergone renal transplantation), one had renal insufficiency, and one proteinuria. Proteinuria disappeared in 4 patients who were proteinuric (2 with nephrotic syndrome) at baseline, and their renal function remained normal. All the live patients had completed at least the 9 years of compulsory education, and 4 had academic degrees. Two female patients had delivered healthy children. CONCLUSION: The outcome of JIA-associated amyloidosis is poor. However, renal disease regressed in some patients under vigorous treatment. Successful treatment makes an active life possible for these patients. PMID- 18381780 TI - Association of CD24 gene polymorphisms with susceptibility to biopsy-proven giant cell arteritis. AB - OBJECTIVE: To investigate the possible implication of CD24 gene in the genetic predisposition to giant cell arteritis (GCA). METHODS: A total of 120 patients diagnosed with biopsy-proven GCA and 195 ethnically matched controls from the same region were studied. Two putative functional polymorphisms, a C to T coding polymorphism (rs8734) and a TG deletion in the 3' untranslated region (rs3838646) were used as CD24 genetic markers and genotyped using a Taqman 5' allelic discrimination assay. RESULTS: The 2 genetic variants showed statistically significant differences between patients with GCA and controls. The strongest association was observed for the rs3838646 TG/del polymorphism, conferring on the "del" allele an increased risk of GCA genetic susceptibility (odds ratio 1.94, 95% confidence interval 1.15-3.27, p = 0.01). In addition, genotypes carrying the rs3838646 "del" allele showed an increased frequency among GCA patients compared to controls (OR 2.31, 95% CI 1.30-4.1, p = 0.003). For the rs8743, an increased frequency of Val/Val homozygous individuals in patients with GCA compared to controls (OR 6.08, 95% CI 1.50-24.63, p = 0.001) was observed. A high degree of linkage disequilibrium was estimated between the 2 polymorphisms (D' = 0.7) and the C/del haplotype was associated with an increased risk of GCA susceptibility (OR 2.10, 95% CI 1.23-3.60, p = 0.005), whereas the C/TG haplotype showed a protective effect (OR 0.63, 95% CI 0.45-0.87, p = 0.005). CONCLUSION: Our results suggest a potential role for the CD24 gene in the susceptibility to GCA in our population. PMID- 18381781 TI - Early-onset osteoarthritis due to otospondylomegaepiphyseal dysplasia in a family with a novel splicing mutation of the COL11A2 gene. AB - OBJECTIVE: To evaluate an approach to the clinical, radiographic, and molecular diagnosis of an underlying skeletal dysplasia in adults presenting with early onset polyarticular osteoarthritis (OA). METHODS: We identified a family with 2 adults with polyarticular OA and a child with generalized arthralgia. General, musculoskeletal, ocular, and auditory evaluations were undertaken. Investigations included radiographs of symptomatic joints, analysis of serum inflammatory markers and joint fluid, and mutational analyses of the COL11A2 gene. RESULTS: The 3 affected individuals had normal stature, mild mid-face hypoplasia, and hearing impairment, but normal eyes. Radiographs of the affected adults showed severe polyarticular OA but did not reveal diagnostic evidence of an underlying skeletal dysplasia. However, the child's radiographs showed enlarged epiphyses with an advanced bone age. The combination of skeletal, facial, and auditory features together with the absence of ocular features indicated that they had otospondylomegaepiphyseal dysplasia, also known as Stickler syndrome type III. The diagnosis was confirmed by identifying a mutation in the COL11A2 gene that encodes the pre-pro-alpha2(XI) chain of type XI collagen that is involved in type II collagen fibrillogenesis. CONCLUSION: Early-onset polyarticular OA may occur in adults without a known or obvious underlying skeletal dysplasia. This study provides an approach to the diagnosis of an underlying skeletal dysplasia in such individuals. PMID- 18381783 TI - Identification of cytokines that might enhance the promoter activity of HLA-B27. AB - OBJECTIVE: Although the mechanism by which HLA-B27 induces ankylosing spondylitis is unclear, a minimum threshold of transcription is essential, a process controlled at the promoter. Our aim was to scan the effect of a panel of cytokines on the promoter of the HLA-B27 gene over serial timepoints. METHODS: The promoter region of B*2705 gene was cloned into a luciferase reporter, stably transfected into HeLa cells, and used to monitor the serial effect of 25 cytokines. Results of HLA-B27 promoter-reporter assays were compared to those of real-time polymerase chain reactions. RESULTS: After an initial delay, significant activation of the HLA-B27 promoter was observed with tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and IFN-beta. While early response of the HLA-B27 promoter was highest with TNF-alpha and IFN-gamma, ultimately the highest activity was observed with IFN-beta. CONCLUSION: The only promoter of HLA-B alleles studied in the past was that of HLA-B7, and always at only a single fixed timepoint of culture. This is the first study to show that activation of HLA-B27 promoter is a sequential event, and that TNF-alpha and IFN beta are major participants at different timepoints. PMID- 18381782 TI - Prospective assessment of body weight, body composition, and bone density changes in patients with spondyloarthropathy receiving anti-tumor necrosis factor-alpha treatment. AB - OBJECTIVE: To determine the changes in body weight, body composition, and bone density in patients with spondyloarthropathy (SpA) receiving anti-tumor necrosis factor-alpha (TNF-alpha) treatment. METHODS: One hundred six patients with SpA (80 men, 26 women) aged 20-71 years were included in a 2-year prospective open study. Fifty-nine patients received infliximab (3 or 5 mg/kg/infusion each 6 or 8 weeks); and 47 patients received etanercept (25 mg twice a week) because of persistent active disease despite an optimal treatment, according to ASsessments in Ankylosing Spondylitis Working Group criteria. Body weight, total body composition (lean mass, fat mass), and spine and femoral bone mineral density (BMD; dual-energy x-ray absorptiometry) were measured at baseline and at 1 and 2 years. RESULTS: There was a significant increase in body weight after 1 year (2.2 +/- 3.9 kg, i.e., 3.4%; p < 0.0001) and 2 years (2.2 +/- 4.7 kg, 3.5%; p < 0.0001), mostly due to a significant gain in fat mass at 1 year (1.4 +/- 2.6 kg, 12.1%; p < 0.0001) and 2 years (1.5 +/- 3.1 kg, 14.5%, p < 0.0001). Gain in lean mass was also significant at 1 year (0.8 +/- 2.2 kg, 1.9%; p < 0.0001) and 2 years (0.9 +/- 2.5 kg, 2%; p < 0.0001). At 2 years, lumbar spine and femur BMD increased: +5.8 +/- 13% (p < 0.0001) and +2.26 +/- 4.5% (p = 0.001), respectively. CONCLUSION: This 2-year prospective study showed a significant increase in body weight at 1 year and 2 years, mostly due to a gain in fat mass and a significant increase in BMD, in patients with SpA receiving anti-TNF-alpha treatment. PMID- 18381784 TI - Human leukocyte antigen and clinical and demographic characteristics in psoriatic arthritis and psoriasis in Chinese patients. AB - OBJECTIVE: Psoriasis and psoriatic arthritis (PsA) are interrelated disorders. To date, no study has compared the differences of genes between patients with PsA and psoriasis and healthy controls in a Chinese population. We conducted a retrospective study to determine the human leukocyte antigen (HLA) -A, -B, -Cw, DR, and -DQ alleles in Chinese patients with PsA and psoriasis. METHODS: HLA studies were performed using polymerase chain reaction sequence-specific oligonucleotide (PCR-SSO) genotyping methods in 91 patients with PsA and 80 with psoriasis and 75 controls. Age at disease onset, sex, disease duration, enthesitis, and uveitis were also analyzed. RESULTS: Among the patients with PsA and psoriasis, the frequency of HLA-B27 was significantly higher in PsA and HLA A*30, -Cw*06, -DR*07 in psoriasis compared with controls. In contrast, HLA-B*58 was more common in controls than in PsA and psoriasis groups, and the prevalence of HLA-DR*17 was significantly higher in controls than in those with psoriasis. Comparing PsA and psoriasis, the prevalence of HLA-B*27 and HLA-Cw*12 were more common in PsA patients, while the prevalence of HLA-DR*07 was higher in those with psoriasis (p < 0.05). Among PsA patients, the association between HLA-B*27 and axial joint involvement and uveitis was significant (p < 0.05). CONCLUSION: Certain HLA alleles are found in Chinese patients with psoriasis (HLA-A*30, Cw*06, -DR*07) and PsA (HLA-B*27). Psoriasis patients with the HLA-B*27 and/or Cw*12 may have higher risk of developing PsA. Ours is the first study to assess the genetic role of HLA in patients with psoriasis and PsA in a Chinese population. PMID- 18381785 TI - Subclinical atherosclerosis in psoriatic arthritis: a case-control study. AB - OBJECTIVE: To investigate the prevalence of subclinical atherosclerosis among patients with psoriatic arthritis (PsA). METHODS: Forty patients with PsA were enrolled. Controls were matched by age, sex, and atherosclerotic risk factors. All patients and controls underwent duplex scan of the carotid arteries. Carotid intima-media thickness (IMT) was evaluated and the presence of atherosclerotic plaques was recorded. The plaques were graded and carotid plaque index was calculated. RESULTS: Patients with PsA had a higher IMT (mean +/- standard deviation, 1.04 +/- 0.35 mm vs 0.88 +/- 0.29 mm in controls; p = 0.03), and had a higher carotid plaque index than did matched controls (2.3 +/- 2.6, compared to 1.12 +/- 2.09; p = 0.03). Multivariate analysis demonstrated that PsA status as well as age and triglyceride levels were associated with the presence of carotid plaque. Other traditional risk factors were more prevalent among patients with PsA; however, they were not statistically significant. CONCLUSION: Our study demonstrates that patients with PsA may have an increased prevalence of subclinical atherosclerosis. These findings may not be solely attributable to traditional risk factors alone. Special attention and strict control of atherosclerotic risk factors in patients with PsA is warranted. PMID- 18381786 TI - Two-year efficacy and safety of infliximab treatment in patients with active psoriatic arthritis: findings of the Infliximab Multinational Psoriatic Arthritis Controlled Trial (IMPACT). AB - OBJECTIVE: To investigate longterm efficacy/safety of infliximab over 2 years in patients with active psoriatic arthritis (PsA). METHODS: Initially, 104 patients were randomized to receive blinded infusions of infliximab 5 mg/kg or placebo at Weeks 0, 2, 6, and 14. At Week 16, all patients received infliximab 5 mg/kg every 8 weeks through Week 46. Seventy-eight of the 87 patients completing the first year continued into the open-label longterm extension and received infliximab 5 mg/kg at Weeks 54, 62, 70, 78, 86, and 94. The primary efficacy endpoint for the study extension was the proportion of patients with at least 20% improvement in the American College of Rheumatology response criteria (ACR20) at Week 98. Radiographic progression was assessed by the PsA-modified van der Heijde-Sharp score in patients with radiographs available at baseline and Week 98 (n = 43). RESULTS: At Week 98, 62% (48/78) of infliximab-treated patients achieved an ACR20 response; 45% (35/78) and 35% (27/78) of patients achieved ACR50 and ACR70 responses, respectively. Among patients with baseline Psoriasis Area and Severity Index scores >or= 2.5, 64% (16/25) achieved > 75% improvement from baseline to Week 98. The average estimated annual radiographic progression with infliximab treatment was significantly reduced versus the estimated baseline rate of progression. No new safety issues were observed during the second year of the study. CONCLUSION: Therapy with infliximab 5 mg/kg through Week 94 produced sustained improvement in joint and skin symptoms, inhibited radiographic progression, and continued to exhibit a favorable benefit-risk ratio in this population with treatment-refractory PsA. PMID- 18381787 TI - Risks and benefits of tumor necrosis factor-alpha inhibitors in the management of psoriatic arthritis: systematic review and metaanalysis of randomized controlled trials. AB - OBJECTIVE: To evaluate the efficacy and safety of tumor necrosis factor-alpha (TNF-alpha) inhibitors in the management of psoriatic arthritis (PsA). METHODS: We searched electronic databases to identify randomized controlled trials (RCT) of adalimumab, etanercept, and infliximab used in patients with PsA. Random effects metaanalysis was undertaken to produce pooled estimates of the relative risk, risk difference, or the weighted mean difference for efficacy and safety outcomes using Stata version 9.0. RESULTS: Six RCT met the inclusion criteria, including 982 patients. All 3 TNF-alpha inhibitors were significantly more effective than placebo on the basis of Psoriatic Arthritis Response Criteria (PsARC) and American College of Rheumatology response criteria ACR20, ACR50, and ACR70 ratings. There were no significant differences between TNF-alpha inhibitors and placebo in the proportions of patients experiencing withdrawal for any reason (RR 0.48, 95% CI 0.20-1.18), or withdrawal due to adverse events (RR 2.14, 95% CI 0.73-6.27), serious adverse events (RR 0.98, 95% CI 0.55-1.77), or upper respiratory tract infections (RR 0.91, 95% CI 0.65-1.28). Pooled rates for injection site reactions were significantly higher for adalimumab and etanercept than for placebo (RR 2.48, 95% CI 1.16-5.29), but there was no significant difference in the proportion of patients experiencing infusion reactions with infliximab (RR 1.03, 95% CI 0.48-2.20) compared against placebo. Indirect analysis did not demonstrate any significant differences between the TNF-alpha inhibitors. CONCLUSION: TNF-alpha inhibitors are effective treatments for PsA with no important added risks associated with their short-term use. There is still a need for longterm risk-benefit assessment of using these drugs for the management of PsA. PMID- 18381788 TI - Diagnostic and prognostic significance of measuring antibodies to alpha-fodrin compared to anti-Ro-52, anti-Ro-60, and anti-La in primary Sjogren's syndrome. AB - OBJECTIVE: To compare sensitivity and specificity of autoantibodies to alpha fodrin with conventional anti-Ro and anti-La antibodies in patients with primary Sjogren's syndrome (pSS). Data on internal organ manifestations were correlated with presence of autoantibodies. METHODS: We collected clinical and laboratory data from 321 patients with pSS (Copenhagen criteria), of which 205 fulfilled the new American-European 2002 consensus criteria. Sera were tested for autoantibodies against alpha-fodrin and recombinant Ro-52, Ro-60, and La proteins. RESULTS: Antibodies to alpha-fodrin were not diagnostically superior to conventional anti-Ro/La testing. IgG anti-La had the highest specificity (97%). A highly significant association was found between presence of anti-La and internal organ manifestations (OR 6, 95% CI 2.99-12.03) or hematological abnormalities. The pattern of autoantibodies was relatively independent of disease duration, indicating that these antibodies appeared early in pSS, probably even years before the first symptoms were manifest. CONCLUSION: We could not confirm that antibodies to alpha-fodrin had higher specificity or sensitivity than anti-Ro/La. Anti-La antibodies were strongly correlated to organ involvement and cytopenias, and thus could serve as a prognostic marker in pSS. PMID- 18381789 TI - Single-nucleotide polymorphisms in CCL2 gene are not associated with susceptibility to systemic sclerosis. AB - OBJECTIVE: To validate the reported association between CC chemokine ligand 2 (CCL2) -2518 G single nucleotide polymorphism and systemic sclerosis (SSc) in a much larger cohort of patients. We also performed subgroup analysis to test the hypothesis that CCL2 variants predispose to specific disease phenotypes. METHODS: Ninety-four Caucasian patients with SSc and 102 matched controls were genotyped by sequence-specific primers-polymerase chain reaction (SSP-PCR) methodology. RESULTS: Six biallelic single-nucleotide polymorphisms (SNP) were investigated (3 in the promoter region, 2 in the exon-coding sequence, and 1 in the 3x untranslated region), in addition to the known functional -2518 (A/G) variant. Six major haplotypes were constructed across all 7 SNP positions. No significant differences in genotype, allele, or haplotype frequency were observed between patients and controls or within disease subgroups. CONCLUSION: Genetic polymorphisms within CCL2 gene are associated with susceptibility neither to SSc nor to specific disease phenotypes. PMID- 18381790 TI - Ethnic disparities among patients with systemic lupus erythematosus in South Carolina. AB - OBJECTIVE: To evaluate whether ethnic disparities in mortality exist among hospitalized patients with systemic lupus erythematosus (SLE) in South Carolina, USA. METHODS: Administrative data were obtained on all SLE patients (ICD-9 code 710.0) hospitalized in South Carolina between 1996 and 2003. An SLE-specific comorbidity index validated as a predictor of hospital mortality was used as a measure of overall comorbidity. Cox proportional hazards models were used to compare mortality rates between Caucasians and African Americans. Post-hoc analyses focused on determining whether disparities were present across different risk strata. RESULTS: Of 6521 hospitalized patients with SLE (5728 female, 793 male), 1280 (19.6%) died. Annual mortality rates were 21.9% among Caucasians and 25.0% among African Americans. The comorbidity index score was significantly higher among African Americans [median 2.0 (interquartile range 0.0-4.0)] versus Caucasians [median 0.0 (IQR 0.0-3.0); p < 0.0001, Wilcoxon rank-sum test]; however, even after multivariate adjustment, African Americans had a 15% increased mortality risk (hazard ratio 1.15, 95% CI 1.02-1.29, p = 0.013). The disparity was strongest among those with less comorbidity (HR 1.39, 95% CI 1.05 1.81, p = 0.017). Among patients with low comorbidity index scores (n = 3485), the annual mortality rate was 8.1% among Caucasians and 9.7% among African Americans. No ethnic differences in mortality were seen for patients with higher comorbidity. CONCLUSION: In South Carolina, ethnic disparities in SLE mortality exist, predominantly among those with the least illness severity. Studies are planned to help clarify whether access and quality of care play a role. PMID- 18381791 TI - Fulvestrant (Faslodex), an estrogen selective receptor downregulator, in therapy of women with systemic lupus erythematosus. clinical, serologic, bone density, and T cell activation marker studies: a double-blind placebo-controlled trial. AB - OBJECTIVE: Estrogen plays a role in the activation of systemic lupus erythematosus (SLE) and in upregulating intracellular signals by binding to the estrogen receptor(s). Fulvestrant (Faslodex, AstraZeneca Pharmaceuticals, Wilmington, DE, USA), an estrogen selective receptor downregulator, competes for receptor binding in vitro and inhibits estrogen action in target cells. We evaluated the efficacy, side effects, and expression of T cell activation markers, following the administration of fulvestrant or placebo to premenopausal patients with SLE. METHODS: Twenty women with moderate SLE Disease Activity Index (SLEDAI; 7.87 +/- 3.7) were enrolled. They were premenopausal with regular menstrual cycles and not taking exogenous hormones. The study was double-blind and placebo-controlled. Ten patients received 250 mg fulvestrant intramuscularly for 12 months, and 10 received the placebo. All were observed monthly and 3 months after final fulvestrant/placebo injection. Measures studied were monthly SLEDAI scores, routine and serologic markers for lupus, and serum concentrations of estrogen and fulvestrant. Expression of T cell calcineurin and CD154 mRNA in peripheral T cells was measured by polymerase chain reaction. Medications the patients were taking were recorded each visit. Bone density was obtained at baseline and at visit 12. RESULTS: Sixteen patients completed the 15-month study, 8 from each group. SLEDAI improved significantly in the fulvestrant group at both 12 months (p = 0.02) and 15 months (p = 0.002), but serologic markers, routine laboratory tests, and bone density did not. Serum estrogen levels were higher in the fulvestrant group and dropped when fulvestrant was discontinued; these differences were not statistically significant. Medications for therapy of lupus to the fulvestrant group were reduced, whereas the placebo group medications were unchanged or increased. Comparison of relative values at individual timepoints revealed significantly lower median values for the T cell activation markers CD154 (p < 0.001) and calcineurin (p = 0.013) in the fulvestrant arm. CONCLUSION: Blocking estrogen receptors in vivo by an estrogen selective receptor downregulator could be considered as a new and relatively safe therapeutic approach in the management of SLE patients with moderately active disease for the 1-year study period. PMID- 18381792 TI - High prevalence of unemployment in patients with systemic lupus erythematosus: association with organ damage and health-related quality of life. AB - OBJECTIVE: To evaluate employment status in relation to organ damage, demographic factors, and health-related quality of life (HRQOL) in patients with systemic lupus erythematosus (SLE). METHODS: One hundred forty-seven patients with SLE, aged 18 to 64 years, were included. Examination of self-reported HRQOL was performed using the Medical Outcome Study Short Form-36 (SF-36). Demographic factors, disease characteristics, and HRQOL were compared between patients with and without paid employment using nonparametric tests. RESULTS: In 147 patients with SLE (mean age 38.4 +/- 11 yrs, mean disease duration 6.3 +/- 6.5 yrs), we found a high rate of unemployment (59%), whereas 93% of patients had previously worked. In patients who had stopped or reduced working, 75% attributed their withdrawal from work at least in part to disease-related factors. Unemployed patients had a significantly higher median age at disease onset, a higher frequency of neuropsychiatric organ damage and diabetes mellitus, and lower HRQOL than employed patients. CONCLUSION: We found a high rate of unemployment (59%) in patients with SLE. Higher age at disease onset, neuropsychiatric organ damage, diabetes mellitus, and reduced HRQOL were associated with unemployment in patients with SLE. Our findings highlight the need to develop strategies to reduce work loss in patients with SLE, for both individual and socioeconomic reasons. PMID- 18381793 TI - Usefulness of enzyme-linked immunospot assay (Elispot) compared to tuberculin skin testing for latent tuberculosis screening in rheumatic patients scheduled for anti-tumor necrosis factor treatment. AB - OBJECTIVE: Treatment with tumor necrosis factor (TNF) antagonists in patients with rheumatic diseases has been associated with increased rates of tuberculosis due to reactivation of latent Mycobacterium tuberculosis (MTb) infection (LTBI). Diagnosis of LTBI is based mainly on the tuberculin skin test (TST), which has certain limitations. METHODS: We compared the TST with an enzyme-linked immunospot interferon-gamma (IFN-gamma) release assay (Elispot; T SPOT TB) for the diagnosis of LTBI in 70 patients with various rheumatic diseases starting treatment with anti-TNF agents. All patients underwent a standard initial evaluation for LTBI including clinical examination, chest radiograph, and standard TST. Freshly isolated peripheral blood mononuclear cells were stimulated ex vivo with MTb-specific antigens (ESAT-6 and CFP10), and IFN-gamma-producing cells were counted (Elispot assay). RESULTS: Twenty-seven patients (38.6%) were TST+ and 16 were Elispot+ (22.8%). The overall level of agreement between the 2 tests was 72.8%, being much higher in patients who were TST- (39/43, 90.6%) than in those who were TST+ (12/27, 44.4%). Discordant results were observed in 19 patients (27.1%). Among TST- patients (n = 43), 4 were Elispot+ (9.3%); we also identified 15 Elispot- patients among 27 TST+ patients (55.6%). Multivariate analysis showed that a history of bacillus Calmette-Guerin (BCG) vaccination was associated with TST+/Elispot- discordant results (p = 0.01), whereas steroid use was linked to TST-/Elispot+ discordant results (p = 0.04). CONCLUSION: Elispot assay is a useful test for diagnosis of LTBI in rheumatic patients scheduled for anti-TNF therapy and identification of patients with false-positive TST results due to previous BCG vaccination. PMID- 18381794 TI - Methotrexate (MTX) pathway gene polymorphisms and their effects on MTX toxicity in Caucasian and African American patients with rheumatoid arthritis. AB - OBJECTIVE: Research has examined the association of folate-dependent gene polymorphisms with methotrexate (MTX) toxicity in racially homogenous patients with rheumatoid arthritis (RA). We examined the influence of MTX transporter gene polymorphisms on MTX toxicity in 2 racial groups of patients with RA. METHODS: Using a retrospective cross-validation approach, the association of polymorphisms in 6 genes in the MTX cellular pathway with MTX toxicity was examined in training and validation cohorts. The genes analyzed were ATP-binding cassette transporter B1 (ABCB1), C1 (ABCC1), C2 (ABCC2), folylpolyglutamyl synthase (FPGS), methylenetetrahydrofolate reductase (MTHFR), and thymidylate synthase (TYMS). Both cohorts included Caucasian Americans and African Americans. Statistical analyses consisted of Fisher exact tests, multivariable logistic regression models, and survival analyses. RESULTS: Four of 25 variants displayed significant associations with MTX toxicity in the training cohort. The intronic single nucleotide polymorphism (SNP) ABCC2 IVS 23+56 T --> C was associated with alopecia in Caucasians (p = 0.035). ABCB1 1236 C --> T was associated with overall toxicity (p = 0.013); ABCC2 1249 G --> A with gastrointestinal toxicity (p = 0.009); and ABCC2 1058 G --> A with hepatotoxicity (p = 0.04) in African Americans. These 4 SNP and the MTHFR 677 C --> T variant were assessed in the validation cohort. Of these, only the MTHFR 677 C --> T SNP was associated with alopecia, and only in African Americans (p = 0.032). The ABCC2 IVS 23+56 T --> C genotype influenced toxicity-related time to discontinuation or dose decrease in the Caucasian validation cohort (p < 0.0001). CONCLUSION: In addition to SNP in folate-dependent genes, MTX transporter gene SNP may be important markers of MTX toxicity in RA. Such pharmacogenetic associations are race-specific. PMID- 18381795 TI - Cyclooxygenase-2 polymorphisms and risk of rheumatoid arthritis in Koreans. AB - OBJECTIVE: To determine the association of single-nucleotide polymorphisms (SNP) in the cyclooxygenase-2 (COX-2) gene with the risk and radiologic severity of rheumatoid arthritis (RA) in Koreans. METHODS: Sequencing of the COX-2 gene using a DNA analyzer revealed genetic variants in 24 Korean DNA samples. A total of 1201 Korean patients with RA and 973 controls were genotyped using the TaqMan method. HLA-DRB1 was genotyped by polymerase chain reaction and sequence-specific oligonucleotide probe hybridization techniques. Logistic regression models were used to calculate odds ratios (OR) and 95% confidence intervals (95% CI) and the corresponding probability values for each SNP site and haplotype. RESULTS: Direct sequencing identified 23 SNP of COX-2 gene, from which 2 common SNP (-1329A-->G and 6365T-->C) were selected based on the linkage disequilibrium status among SNP and minor allele frequencies. The -899G-->C SNP was also studied because it is reportedly associated with the risk of RA. The -1329A-->G SNP was not significantly associated with the risk of RA. However, the risk of RA was significantly lower in the presence of the C allele for 6365T-->C (OR 0.50, 95% CI 0.29-0.85, in a recessive model, and OR 0.80, 95% CI 0.67-0.97, in a codominant model). The C allele for -899G-->C was also associated with a significantly lower risk of RA (OR 0.67, 95% CI 0.48-0.95, in a codominant model). The radiologic severity of RA was not associated with COX-2 polymorphisms. CONCLUSION: Our study revealed a possible protective influence of the C allele for 6365T-->C and for -899G-->C in RA. PMID- 18381796 TI - The functional variant (Asp299gly) of toll-like receptor 4 (TLR4) influences TLR4 mediated cytokine production in rheumatoid arthritis. AB - OBJECTIVE: To investigate functional consequences of the Toll-like receptor 4 (TLR4) variant (Asp299Gly) in rheumatoid arthritis (RA). METHODS: Peripheral blood mononuclear cells from 28 patients with RA carrying or not carrying the TLR4 variant were incubated with lipopolysaccharide (LPS) and heat shock protein B8 (HSPB8). Concentrations of interleukin 6 (IL-6), tumor necrosis factor alpha(TNF-alpha), and IL-10 were determined along with TLR4 and CD14 expression. RESULTS: TLR4 expression was similar in patients carrying or not carrying the variant. In contrast, both LPS and HSPB8 resulted in significantly lower secretion of IL-6, TNF-alpha, and IL-10 in those who carried the variant, whereas the frequency of CD14+ cells was higher in these individuals. CONCLUSION: TLR4 variant clearly reduces its potency to mediate signaling. Correction for CD14+ cells is necessary in comparable experiments. PMID- 18381797 TI - Prevalence and distribution of autoimmune diseases in 368 rheumatoid arthritis families. AB - OBJECTIVE: To investigate whether frequency of rheumatoid arthritis (RA) and/or other autoimmune (AI) disorders was increased in RA French Caucasian families among the first- (FDR) and second-degree relatives (SDR), and to test whether the presence of AI disease family history identified a specific RA subset. METHODS: We conducted telephone interviews to obtain histories of AI diseases among the FDR and SDR of 368 RA probands, belonging either to trio or affected sib-pair (ASP) families. All the AI diagnoses were confirmed by the physician of the affected relative. RESULTS: Probands of the ASP families were characterized by older age at RA onset, longer disease duration, and larger family size versus trio families. In the trio families, the prevalence of AI diseases was 6.05% (4.76%-7.57%) in FDR and 2.40% (1.85%-3.06%) in SDR. In ASP families, the prevalence of AI diseases was, respectively, 10.24% (8.68%-11.97%) and 1.79% (1.41%-2.25%). The most frequent AI diseases among relatives were RA, thyroid AI diseases, and vitiligo. In trio families, a proband with a mean age of RA onset < 30 years was associated with AI disease prevalence in the relatives, and male gender was associated with prevalence of RA among the FDR. CONCLUSION: The prevalence of AI diseases is increased, particularly among FDR, in French RA families, and some characteristics of the RA proband seem to be associated with prevalence of AI diseases in families. PMID- 18381798 TI - Serum antibodies against intact human collagen IX are elevated at onset of rheumatoid arthritis but are not related to development of erosions. AB - OBJECTIVE: To measure the presence of autoantibodies binding to intact human recombinant collagen IX and assess their usefulness as a diagnostic marker and an indicator of disease activity in rheumatoid arthritis (RA). METHODS: Recombinant human full-length collagen IX (rCIX) was produced in a baculovirus expression system and purified for use in ELISA developed to detect antibodies to native and denatured collagen IX. Fifty-three patients with recent-onset rheumatoid factor seropositive RA were analyzed for the presence of rCIX antibodies of the IgG type at the time of initial diagnosis and after 3, 6, 12, and 24 months of followup. The RA sera were accompanied by 30 controls. Associations were determined between patients' antibody titers, development of erosions in the hands and feet, and various clinical and laboratory markers. RESULTS: Serum antibody levels among patients with RA at time of diagnosis were 1.78 times higher against native rCIX (p < 0.001) and 1.71 times higher against denatured rCIX (p < 0.001) than in the controls, and they remained high during the followup. No correlation was seen between antibody levels and clinical and laboratory findings. CONCLUSION: Our data show that patients with recent-onset RA have significantly elevated levels of autoantibodies to human rCIX. These autoantibodies were observed already at the early stages of the disease, which may reflect their diagnostic potential in RA. PMID- 18381799 TI - BAT1 promoter polymorphism is associated with rheumatoid arthritis susceptibility. AB - OBJECTIVE: To analyze whether the polymorphisms -22 (G/C) and -348 (C/T) of the BAT1 gene are associated with susceptibility to rheumatoid arthritis (RA). METHODS: One hundred fifty-six patients with RA and 154 controls were genotyped for HLA-DRB1 and the polymorphisms -22 and -348 of the BAT1 gene. RESULTS: HLA DRB1*04 alleles were associated with RA susceptibility (33.9% vs 20.1%; pc = 0.04). Among these, HLA-DRB1*0401 (13.4% vs 5.1%; pc = 0.04) and HLA-DRB1*0404 (5.7% vs 1.2%; pc = 0.2) were increased in patients with RA. Additionally, carriage of BAT1 -348T polymorphism was strongly associated with RA (23.7% vs 12.1%; pc = 0.0002). Significantly, BAT1 -348T was in linkage disequilibrium with HLA-DRB1*0404 and HLA-DRB1*0405. However, BAT1 -348 T was associated independently with HLA-DRB1 shared-epitope alleles (42.6% vs 18.9%; p = 0.001). CONCLUSION: The BAT1 -348T polymorphism is associated with RA susceptibility independently of HLA-DRB1. The role of BAT1 in the regulation of tumor necrosis factor-a suggests that BAT1 may regulate the inflammatory response observed in patients with RA. PMID- 18381800 TI - The measurement and analysis of age-related changes in Caenorhabditis elegans. AB - Aging is characterized by progressive degenerative changes in tissue organization and function that increase the probability of mortality. Major goals of aging research include elucidating the series of events that cause degenerative changes and analyzing environmental and genetic factors that modulate these changes. The basis for mechanistic studies of aging are accurate and precise descriptions of age-related changes, since these descriptions define the aging phenotype. Here we review studies that describe age-related changes in C. elegans including measurements of integrated functions such as behavior, microscopic analyses of tissue organization, and biochemical studies of macromolecules. Genetic and environmental factors that influence these changes are described, and studies that analyze the relationships between different age-related changes are discussed. Together these studies provide fundamental insights into aging in C. elegans that may be relevant to aging in other animals. PMID- 18381801 TI - A synthetic Toll-like receptor 2 ligand decreases allergic immune responses in a mouse rhinitis model sensitized to mite allergen. AB - It has been proposed that activation of Toll-like receptors (TLRs) plays crucial roles in the polarization of adaptive immune responses. A synthetic Toll-like receptor 2 (TLR2) ligand, Pam3CSK4, has been reported to modulate the balance of Th1/Th2 responses. We evaluated the modulation effect of Pam3CSK4 on allergic immune response in a mouse rhinitis model sensitized to house dust mite allergen (HDM). Mice were sensitized and challenged with Dermatophagoides farinae allergen (Der f), and then the allergic mice were treated by Pam3CSK4. Nasal allergic symptoms and eosinophils were scored. Der f-specific cytokine responses were examined in the splenocytes and bronchoalveolar lavage fluid (BALF). Serum level of total IgE was also detected. After establishing a mouse allergic rhinitis model with HDM, we have showed that Pam3CSK4 treatment not only ameliorated the nasal allergic symptoms remarkably but also decreased the eosinophils and total inflammation cells in BALF significantly. Analysis of cytokine profile found that IFN-gamma released from either BALF or stimulated splenocytes increased markedly in Pam3CSK4-treated mice, while IL-13 decreased significantly. Moreover, serum level of total IgE was significantly lower in Pam3CSK4-treated mice than in the untreated. Thus, in an allergic rhinitis mouse model developed with HDM, Pam3CSK4 was shown to exhibit an antiallergic effect, indicating its potential application in allergic diseases. PMID- 18381802 TI - Proteiomic patterns for endometrial cancer using SELDI-TOF-MS. AB - Serum samples from endometrial cancer (EC) patients and healthy females were analyzed using surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF-MS) to discover the potential diagnostic biomarker for detection of EC. A preliminary training set of spectra derived from 40 EC patients and 30 healthy women were used to develop a proteomic model that effectively discriminated cancer patients from healthy women. The training set had a specificity of 100% and sensitivity of 92.5% in the EC detection. A blind test set, including 20 new cancer cases and 10 healthy women, was used to validate the sensitivity and specificity of this multivariate model, which had a corresponding results of 60% in specificity and 75% in sensitivity, respectively. The combination of SELDI-TOF-MS with bioinformatics tools could help find new biomarkers and establish the detection of EC with high sensitivity and specificity. PMID- 18381803 TI - Using oligonucleotide suspension arrays for laboratory identification of bacteria responsible for bacteremia. AB - The aim of this study was to develop and validate an oligonucleotide suspension array for rapid identification of 15 bacterial species responsible for bacteremia, particularly prevalent in Chinese hospitals. The multiplexed array, based on the QIAGEN LiquiChip Workstation, included 15 oligonucleotide probes which were covalently bound to different bead sets. PCR amplicons of a variable region of the bacterial 23S rRNA genes were hybridized to the bead-bound probes. Thirty-eight strains belonging to 15 species were correctly identified on the basis of their corresponding species-specific hybridization profiles. The results show that the suspension array, in a single assay, can differentiate isolates over a wide range of strains and species, and suggest the potential utility of suspension array system to clinical laboratory diagnosis. PMID- 18381804 TI - Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein. AB - OBJECTIVE: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP. METHODS: The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP. RESULTS: After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently. CONCLUSION: We have successfully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies. PMID- 18381805 TI - A preliminary study on the teratogenesis of dexamethasone and the preventive effect of vitamin B12 on murine embryonic palatal shelf fusion in vitro. AB - Excessive dexamethasone (Dex) administrated into pregnant mice during critical periods of palatal development can produce a high incidence of cleft palate. Its mechanisms remain unknown. Vitamin B12 has been shown to antagonize the teratogenic effects of Dex, which, however, remains controversial. In this study, we investigated the effects of Dex and vitamin B12 on murine embryonic palatal shelf fusion using organ culture of murine embryonic shelves. The explanted palatal shelves on embryonic day 14 (E14) were cultured for 24, 48, 72 or 96 h in different concentrations of Dex and/or vitamin B12. The palatal shelves were examined histologically for the morphological alterations on the medial edge epithelium (MEE) and fusion rates among different groups. It was found that the palatal shelves were not fused at 72 h or less of culture in Dex group, while they were completely fused in the control and vitamin B12-treated groups at 72 and 96 h, respectively. The MEE still existed and proliferated. In Dex+vitamin B12 group the palatal shelves were fused at each time point in a similar rate to controls. These results may suggest that Dex causes teratogenesis of murine embryonic palatal shelves and vitamin B12 prevents the teratogenic effect of Dex on palatogenesis on murine embryos in vitro. PMID- 18381806 TI - Resistin does not down-regulate the transcription of insulin receptor promoter. AB - OBJECTIVE: To detect the effect of resistin on the transcription of insulin receptor promoter. METHODS: Luciferase reporter gene was fused downstream of human insulin receptor promoter and the enzymatic activity of luciferase was determined in the presence or absence of resistin. The resistin expressed with plasmid was stained with antibody against Myc tag which was in frame fused with resistin coding sequence, and then imaged with confocal microscopy. RESULTS: The treatment of pIRP-LUC transfected cells with recombinant resistin did not result in significant difference in the enzymatic activity of luciferase compared to the untreated cells. Cell staining showed that green fluorescence could be observed in the cytoplasm, but not in the nucleus. CONCLUSION: The results suggest that the endogenous resistin may functionally locate in the cytoplasm, but does not enter the nucleus and not down-regulate the transcription of insulin receptor promoter. PMID- 18381807 TI - Drainage after total thyroidectomy or lobectomy for benign thyroidal disorders. AB - OBJECTIVE: This prospective randomized clinical trial was conducted to evaluate the necessity of drainage after total thyroidectomy or lobectomy for benign thyroidal disorders. METHODS: A total of 116 patients who underwent total thyroidectomy or lobectomy for benign thyroidal disorders were randomly allocated to be drained or not. Operative and postoperative outcomes including operating time, postoperative pain assessed by visual analogue scale (VAS), total amount of intramuscular analgesic administration, hospital stay, complications, necessity for re-operation and satisfaction of patients were all assessed. RESULTS: The mean operating time was similar between two groups (the drained and non-drained groups). The mean VAS score was found to be significantly low in the non-drained group patients in postoperative day (POD) 0 and POD 1. The mean amount of intramuscular analgesic requirement was significantly less in the non-drained group. One case of hematoma, two cases of seroma and three cases of transient hypoparathyroidism occurred in the non-drained group, whereas one case of hematoma, two cases of seroma, two cases of wound infections and two cases of transient hypoparathyroidism occurred in the drained group. No patient needed re operation for any complication. The mean hospital stay was significantly shorter and the satisfaction of patients was superior in the non-drained group. CONCLUSION: These findings suggest that postoperative complications cannot be prevented by using drains after total thyroidectomy or lobectomy for benign thyroid disorders. Furthermore, the use of drains may increase postoperative pain and the analgesic requirement, and prolong the hospital stay. In the light of these findings, the routine use of drains might not be necessary after thyroid surgery for benign disorders. PMID- 18381808 TI - Changes of tear film and tear secretion after phacoemulsification in diabetic patients. AB - OBJECTIVE: To evaluate tear film stability and tear secretion in patients with diabetes after phacoemulsification. METHODS: Twenty-five diabetic cataract patients and 20 age-matched non-diabetic cataract patients as control underwent phacoemulsification. Tear film break-up time (TFBUT), Schirmer I test (SIT), corneal fluorescein staining, and dry eye symptoms were measured pre- and postoperatively. RESULTS: Diabetics had a decreased preoperative TFBUT and SIT. TFBUT was reduced on Day 1 and recovered on Day 180 postoperatively in both groups. SIT was increased after phacoemulsification, but returned to preoperative levels by Day 180 in non-diabetics, whereas it was lower than preoperative level in diabetics. Positive corneal fluorescein staining was elevated in both groups, and returned to preoperative levels only in controls. Dry eye symptoms were similar to fluorescein staining in both groups. CONCLUSION: Tear secretion was reduced in diabetic cataract patients after phacoemulsification, which worsened dry eye symptoms and predisposed those patients to ocular damage. PMID- 18381809 TI - Values of endoscopic ultrasonography for diagnosis and treatment of duodenal protruding lesions. AB - OBJECTIVE: The diagnoses of patients with duodenal protruding lesions are difficult when using conventional examinations such as computed tomography (CT) and conventional endoscope etc. Thus, we investigated the clinical value of endoscopic ultrasonography (EUS) with miniature ultrasonic probes on the diagnosis and treatment of duodenal protruding lesions. METHODS: Patients with duodenal protruding lesions who were indicated for EUS were examined by EUS with 12 approximately 15 MHz miniature ultrasonic probes and double-cavity electronic endoscope. According to diagnosis of EUS, those patients were indicated for biopsy and treatment received biopsy, endoscopic resection or surgical excision. The postoperative histological results were compared with the preoperative diagnosis of EUS. Those patients without endoscopic resection or surgical excision were periodically followed up with EUS. RESULTS: A total of 169 patients with duodenal protruding lesions were examined by EUS, of which 40 were diagnosed with cysts, 36 with inflammatory protruding or polyp, 25 with Brunner's gland adenoma, 19 with ectopic pancreas, 17 with gastrointestinal stromal tumor, 12 with extrinsic compression, 12 with minor papilla, 6 with lipoma, 1 with adenocarcinoma and 1 with lymphoma. After EUS examinations, 75 patients received biopsy, endoscopic resection or surgical excision respectively. The postoperative histological results of 70 patients were completely consistent with the preoperative diagnosis of EUS, with 93.33% diagnostic accuracy. The results of follow-up with EUS indicated that duodenal cyst, Brunner's gland adenoma, ectopic pancreas, gastrointestinal stromal tumor and lipoma remained unchanged within 1 approximately 3 years. No related complications occurred among all patients that received EUS examinations. CONCLUSION: EUS is an effective and reliable diagnostic method for duodenal protruding lesions. PMID- 18381810 TI - Video-assisted thoracoscopic surgery (VATS) for bilateral primary spontaneous pneumothorax. AB - OBJECTIVE: To review our experience of the treatment of bilateral primary spontaneous pneumothorax (PSP) by video-assisted thoracoscopic surgery (VATS). MATERIALS AND METHODS: Retrospective chart review was followed by an on-clinic or telephone interview. Patients were cared for by one thoracic surgeon in four medical centers or community hospitals in Northern and Central Taiwan. Thirteen patients with bilateral PSP underwent bilateral VATS simultaneously or sequentially from July 1994 to December 2005. RESULTS: Twelve males and one female, with age ranging from 15 to 36 years (mean 23.1 years), were treated with VATS for bilateral PSP, under the indications of bilateral pneumothoracis simultaneously (n=4) or sequentially (n=9). The interval between the first and second contra-lateral VATS procedure for non-simultaneous PSP patients ranged from 7 d to 6 years. Eleven of 13 patients (84.6%) had prominent pulmonary bullae/blebs, and underwent bullae resection with mechanical or chemical pleurodesis. The mean operative time was (45.6+/-18.3) min (range 25 approximately 96 min) and (120.6+/-28.7) min (range 84 approximately 166 min) respectively for the non-simultaneous (second VATS for the recurrence of contralateral side after first VATS) and simultaneous (bilateral VATS in one operation) procedures. There was no postoperative mortality. However, prolonged air leakage (>7 d) occurred in one patient (7.7%) who recovered after conservative treatment. The mean duration of chest tube drainage was 3.1 d and the median follow up period was 3.4 years. CONCLUSIONS: VATS is a safe and effective procedure in the treatment of bilateral PSP. Bilateral VATS is only recommended for patients with simultaneously bilateral PSP, because the incidence of recurrence, even with visible bullae, was not so high in my group and in some previous literature. Bilateral VATS in a supine position should only be used in selective cases, because of possible pleural adhesion or hidden bullae on the posterior side. PMID- 18381811 TI - Sodium arsenite reduces severity of dextran sulfate sodium-induced ulcerative colitis in rats. AB - The histopathological features and the associated clinical findings of ulcerative colitis (UC) are due to persistent inflammatory response in the colon mucosa. Interventions that suppress this response benefit UC patients. We tested whether sodium arsenite (SA) benefits rats with dextran sulfate sodium (DSS)-colitis. The DSS-colitis was induced by 5% DSS in drinking water. SA (10 mg/kg; intraperitoneally) was given 8 h before DSS treatment and then every 48 h for 3 cycles of 7, 14 or 21 d. At the end of each cycle rats were sacrificed and colon sections processed for histological examination. DSS induced diarrhea, loose stools, hemoccult positive stools, gross bleeding, loss of body weight, loss of epithelium, crypt damage, depletion of goblet cells and infiltration of inflammatory cells. The severity of these changes increased in the order of Cycles 1, 2 and 3. Treatment of rats with SA significantly reduced this severity and improved the weight gain. PMID- 18381812 TI - No detectable endothelial- or leukocyte-derived L-selectin ligand activity on the endothelium in inflamed cremaster muscle venules. AB - L-selectin is important in mediating leukocyte recruitment in inflammation. The role of L-selectin was for long believed to be influenced by an inducible endothelial ligand; however, L-selectin ligand activity was recently shown to be mediated by leukocytic P-selectin glycoprotein ligand 1 (PSGL-1). Still, it is unknown whether PSGL-1 is deposited on the endothelium or whether leukocyte fragments or leukocytic uropods are presented on the venular surface. Moreover, it is unclear whether ligands for L-selectin other than PSGL-1 are present in inflammation. Overall, this has complicated understanding of the mechanisms that guide recruitment of inflammatory cells. Here, I used intravital microscopy on mouse cremaster muscle venules to show that L-selectin influences leukocyte rolling in inflammation exclusively by mediating L-selectin/PSGL-1-dependent, secondary capture to rolling and adherent leukocytes. I show that leukocyte primary capture in inflammation is mediated almost entirely by P-selectin, whereas the capacity of E-selectin to mediate capture appears to be minimal. In parallel, primary capture remaining after function inhibition of P-selectin is not decreased by blockage or absence of L-selectin. Rolling along the endothelium in venules following a number of inflammatory treatments was abolished by simultaneous blockage of P-selectin, E-selectin, and VCAM-1, indicating that there is no additional adhesive pathway involving L-selectin or any other molecule that can mediate leukocyte rolling in inflamed cremaster muscle venules in response to the used stimuli. Moreover, in vivo staining failed to detect any L-selectin ligand activity on the endothelium. These data demonstrate that expression of L-selectin on leukocytes is insufficient for mediating rolling and efficient recruitment of leukocytes in inflammation. PMID- 18381813 TI - Vascular screening of over 40s will save 2000 lives a year, says health department. PMID- 18381814 TI - Voltage-dependent anion channel 1 is involved in endostatin-induced endothelial cell apoptosis. AB - Endostatin (ES) was reported to stimulate apoptosis in endothelial cells, but the exact mechanism remains controversial. In the present study, we elucidate the mechanism of ES-induced endothelial cell apoptosis. Our results indicate that ES induces cytochrome c release and caspase-9 activation in human microvascular endothelial cells (HMECs) at the concentration of 1 microM for 24 h, which initiates the apoptosis process. Further, ATP production, mitochondrial membrane potential, and tubule formation assays showed that ES promotes the mitochondrial permeability transition pore (mPTP) opening via voltage-dependent anion channel 1 (VDAC1), a major component of mitochondrial outer membrane. Knocking down VDAC1 by small interfering RNA attenuates ES-induced apoptosis, while overexpression of VDAC1 enhances the sensitivity of endothelial cells to ES. Moreover, we reveal that ES induces the reduction of hexokinase 2 (HK2), which, in turn, promotes VDAC1 phosphorylation and accumulation. Data from two-dimensional electrophoresis, immunoprecipitation, mPTP opening, and caspase-3 activation assays indicate that two serine residues of VDAC1, Ser-12 and Ser-103, can modulate VDAC1 protein level and thus the sensitivity to apoptosis stimuli. On the basis of these findings, we conclude that VDAC1 plays a vital role in modulating ES-induced endothelial cell apoptosis. PMID- 18381815 TI - Evidence for cooperative signal triggering at the extracellular loops of the TSH receptor. AB - The mechanisms governing transition of the thyroid stimulating hormone (TSH) receptor (TSHR) from basal to active conformations are poorly understood. Considering that constitutively activating mutations (CAMs) and inactivating mutations in each of the extracellular loops (ECLs) trigger only partial TSHR activation or inactivation, respectively, we hypothesized that full signaling occurs via multiple extracellular signal propagation events. Therefore, individual CAMs in the extracellular region were combined to create double and triple mutants. In support of our hypothesis, combinations of mutants in the ECLs are in some cases additive, while in others they are even synergistic, with triple mutant I486A/I568V/V656F exhibiting a 70-fold increase in TSH-independent signaling. The proximity but likely different spatial orientation of the residues of activating and inactivating mutations in each ECL supports a dual functionality to facilitate signal induction and conduction, respectively. This is the first report for G-protein coupled receptors, suggesting that multiple and cooperative signal propagating events at all three ECLs are required for full receptor activation. Our findings provide new insights concerning molecular signal transmission from extracellular domains toward the transmembrane helix bundle of the glycoprotein hormone receptors. PMID- 18381816 TI - An Adam15 amplification loop promotes vascular endothelial growth factor-induced ocular neovascularization. AB - Proteins with a disintegrin and a metalloproteinase domain (ADAMs) are a family of membrane-bound proteinases that bind integrins through their disintegrin domain. In this study, we have found modest expression of ADAM15 in pericytes in normal retina and strong up-regulation of ADAM15 in retinal vascular endothelial cells in ischemic retina. Increased expression of vascular endothelial growth factor (VEGF) in the retina in the absence of ischemia also increased ADAM15 levels, and knockdown of Vegf mRNA in ischemic retina reduced Adam15 mRNA. Mice deficient in ADAM15 showed a significant reduction in ischemia-induced retinal neovascularization, choroidal neovascularization at rupture sites in Bruch's membrane, and VEGF-induced subretinal neovascularization. ADAM15-deficient mice also showed reduced levels of VEGF(164), VEGF receptor 1, and VEGF receptor 2 in ischemic retina. These data suggest that ADAM15 and VEGF participate in an amplification loop; VEGF increases expression of ADAM15, which in turn increases expression of VEGF and its receptors. Perturbation of the loop by elimination of ADAM15 suppresses ocular neovascularization in 3 different model systems, and thus ADAM15 provides a new therapeutic target for diseases complicated by neovascularization. PMID- 18381817 TI - Water inertial reorientation: hydrogen bond strength and the angular potential. AB - The short-time orientational relaxation of water is studied by ultrafast infrared pump-probe spectroscopy of the hydroxyl stretching mode (OD of dilute HOD in H(2)O). The anisotropy decay displays a sharp drop at very short times caused by inertial orientational motion, followed by a much slower decay that fully randomizes the orientation. Investigation of temperatures from 1 degrees C to 65 degrees C shows that the amplitude of the inertial component (extent of inertial angular displacement) depends strongly on the stretching frequency of the OD oscillator at higher temperatures, although the slow component is frequency independent. The inertial component becomes frequency-independent at low temperatures. At high temperatures there is a correlation between the amplitude of the inertial decay and the strength of the O-D O hydrogen bond, but at low temperatures the correlation disappears, showing that a single hydrogen bond (OD O) is no longer a significant determinant of the inertial angular motion. It is suggested that the loss of correlation at lower temperatures is caused by the increased importance of collective effects of the extended hydrogen bonding network. By using a new harmonic cone model, the experimentally measured amplitudes of the inertial decays yield estimates of the characteristic frequencies of the intermolecular angular potential for various strengths of hydrogen bonds. The frequencies are in the range of approximately 400 cm(-1). A comparison with recent molecular dynamics simulations employing the simple point charge-extended water model at room temperature shows that the simulations qualitatively reflect the correlation between the inertial decay and the OD stretching frequency. PMID- 18381818 TI - Evidence that strong positive selection drives neofunctionalization in the tandemly duplicated polyhomeotic genes in Drosophila. AB - The polyhomeotic (ph) locus in Drosophila melanogaster consists of the two tandemly duplicated genes ph-d (distal) and ph-p (proximal). They code for transcriptional repressors belonging to the Polycomb group proteins, which regulate homeotic genes and hundreds of other loci. Although the duplication of ph occurred at least 25 million to 30 million years ago, both copies are very similar to each other at both the DNA and the protein levels, probably because of the action of frequent gene conversion. Despite this homogenizing force, differential regulation of both transcriptional units suggests that the functions of the duplicates have begun to diverge. Here, we provide evidence that this functional divergence is driven by positive selection. Based on resequencing of an approximately 30-kb region around the ph locus in an African sample of D. melanogaster X chromosomes, we identified a selective sweep, estimated its age and the strength of selection, and mapped the target of selection to a narrow interval of the ph-p gene. This noncoding region contains a large intron with several regulatory elements that are absent in the ph-d duplicate. Our results suggest that neofunctionalization has been achieved in the Drosophila ph genes through the action of strong positive selection and the inactivation of gene conversion in part of the gene. PMID- 18381819 TI - DNA surface hybridization regimes. AB - Surface hybridization reactions, in which sequence-specific recognition occurs between immobilized and solution nucleic acids, are routinely carried out to quantify and interpret genomic information. Although hybridization is fairly well understood in bulk solution, the greater complexity of an interfacial environment presents new challenges to a fundamental understanding, and hence application, of these assays. At a surface, molecular interactions are amplified by the two dimensional nature of the immobilized layer, which focuses the nucleic acid charge and concentration to levels not encountered in solution, and which impacts the hybridization behavior in unique ways. This study finds that, at low ionic strengths, an electrostatic balance between the concentration of immobilized oligonucleotide charge and solution ionic strength governs the onset of hybridization. As ionic strength increases, the importance of electrostatics diminishes and the hybridization behavior becomes more complex. Suppression of hybridization affinity constants relative to solution values, and their weakened dependence on the concentration of DNA counterions, indicate that the immobilized strands form complexes that compete with hybridization to analyte strands. Moreover, an unusual regime is observed in which the surface coverage of immobilized oligonucleotides does not significantly influence the hybridization behavior, despite physical closeness and hence compulsory interactions between sites. These results are interpreted and summarized in a diagram of hybridization regimes that maps specific behaviors to experimental ranges of ionic strength and probe coverage. PMID- 18381820 TI - Microbial carbohydrate depolymerization by antigen-presenting cells: deamination prior to presentation by the MHCII pathway. AB - After uptake by the endosome of an antigen-presenting cell (APC), exogenous proteins are known to be degraded into peptides by protease digestion. Here, we report the mechanism by which pure carbohydrates can be depolymerized within APC endosomes/lysosomes by nitric oxide (NO)-derived reactive nitrogen species (RNSs) and/or superoxide-derived reactive oxygen species (ROSs). Earlier studies showed that depolymerization of polysaccharide A (PSA) from Bacteroides fragilis in the endosome depends on the APC's having an intact inducible nitric oxide synthase (iNOS) gene; the chemical mechanism underlying depolymerization of a carbohydrate within the endosome/lysosome is described here. Examining the ability of the major RNSs to degrade PSA, we determined that deamination is the predominant mechanism for PSA processing in APCs and is a required step in PSA presentation to CD4(+) T cells by MHCII molecules. Structural characterization of the NO derived product PSA-NO indicates that partial deaminative depolymerization does not alter the zwitterionic nature of PSA. Unlike native PSA, PSA-NO is presented by iNOS-deficient APCs to induce CD4(+) T cell proliferation. Furthermore, metabolically active APCs are required for PSA-NO presentation. In contrast to PSA degradation by RNSs, dextran depolymerization in the endosome depends on ROSs, including hydrogen peroxide- and superoxide-derived ROSs. This study provides evidence that MHCII pathway-mediated carbohydrate antigen processing in APCs is achieved by chemical reactions. RNSs and ROSs may be involved in the presentation of glycopeptides by MHC molecules via the processing of other carbohydrate-containing antigens, such as bacterial or viral glycoproteins or glycoconjugate vaccines. PMID- 18381821 TI - CASY-1, an ortholog of calsyntenins/alcadeins, is essential for learning in Caenorhabditis elegans. AB - Calsyntenins/alcadeins are type I transmembrane proteins with two extracellular cadherin domains highly expressed in mammalian brain. They form a tripartite complex with X11/X11L and APP (amyloid precursor protein) and are proteolytically processed in a similar fashion to APP. Although a genetic association of calsyntenin-2 with human memory performance has recently been reported, physiological roles and molecular functions of the protein in the nervous system are poorly understood. Here, we show that CASY-1, the Caenorhabditis elegans ortholog of calsyntenins/alcadeins, is essential for multiple types of learning. Through a genetic screen, we found that casy-1 mutants show defects in salt chemotaxis learning. casy-1 mutants also show defects in temperature learning, olfactory adaptation, and integration of two sensory signals. casy-1 is widely expressed in the nervous system. Expression of casy-1 in a single sensory neuron and at the postdevelopmental stage is sufficient for its function in salt chemotaxis learning. The fluorescent protein-tagged ectodomain of CASY-1 is released from neurons. Moreover, functional domain analyses revealed that both cytoplasmic and transmembrane domains of this protein are dispensable, whereas the ectodomain, which contains the LG/LNS-like domain, is critically required for learning. These results suggest that learning is modulated by the released ectodomain of CASY-1. PMID- 18381822 TI - Payments to injured professional jockeys in British horse racing (1996-2006). AB - OBJECTIVE: To determine the direct and indirect cost of injuries in professional jockeys as a result of race riding. DESIGN: Cohort study. SETTING: Professional horse racing. PARTICIPANTS: Professional jockeys in the UK registered with the Jockey Club. MAIN OUTCOME MEASURES: Payouts under the Professional Riders Insurance Scheme (PRIS) for injuries sustained by professional jockeys, which were recorded by the Jockey Club injury database between 1996 and 2006 inclusive. RESULTS: In the 11-year period of the study, there were 1328 injuries that resulted in jockeys missing a total of 71,509 days of racing, and as a result PRIS paid out 4,496,019 pounds sterling in compensation. Most injuries were minor, with a third of injured jockeys returning to race riding within 2 weeks and 45% of the claims receiving a payout of < 1000 pounds sterling. Almost half the claims were due to fractures, with the clavicle being the most common site of injury. Joint dislocations accounted for the highest insurance payout and the longest time off racing. CONCLUSIONS: Horse racing is a sport known to have a high risk of injury. This study has found that almost 4.5 million pounds sterling was spent on the injuries that were reported to PRIS during the 11-year time period 1996-2006. This study sheds light on the overall injury cost in professional horse racing and will form the basis for more formal health economic analysis. PMID- 18381823 TI - Physical activity throughout life reduces the atherosclerotic wall process in the carotid artery. AB - BACKGROUND: Good cardiorespiratory fitness has been associated with a reduced risk for clinical events of atherosclerotic vascular diseases. It is still unclear how this relates to a slower progression of the early atherosclerosis wall process. METHOD: Using a dynamic model, we generated new parameters for describing the pathologic wall process in the carotid artery, based on an automatic layer detection system. In this study, we scrutinised the influence of two ultrasonographic parameters, intima-media thickness (IMT) and roughness, by comparing two groups: a healthy inactive group (PIP) (mean (SD) age 64.37 (5.10) years; n = 50) and a healthy lifelong physically active group (PA) (mean (SD) age 64.48 (3.45) years; n = 51). All subjects underwent a blood test, spiroergometry, echocardiography and carotid ultrasound examination. RESULTS: There was a significant difference in the well known risk factors for cardiovascular disease (for example, high density lipoprotein cholesterol, triglyceride) between groups. PIP compared to PA had a significantly higher roughness (PIP 0.073 (0.015) vs PA 0.065 (0.0156); p<0.01). No significantly higher IMT was found for PIP (PIP 0.89 (0.18) vs PA 0.90 (0.22); p = 0.63) compared to PA. In this cross sectional study of middle aged men, Vo(2)max was inversely associated with carotid atherosclerotic parameters. CONCLUSION: In this study, good cardiorespiratory fitness was associated with an increase of the proven risk factors and a reduction of atherosclerosis in the common carotid artery. Roughness seems to be significantly more sensitive than IMT for characterising the changes of the arterial wall. We suggest measuring roughness in addition to IMT to gain additional information about the atherosclerotic wall. PMID- 18381824 TI - Doppler ultrasonography of the anterior knee tendons in elite badminton players: colour fraction before and after match. AB - BACKGROUND: Anterior knee tendon problems are seldom reported in badminton players although the game is obviously stressful to the lower extremities. HYPOTHESES: Painful anterior knee tendons are common among elite badminton players. The anterior knee tendons exhibit colour Doppler activity. This activity increases after a match. Painful tendons have more Doppler activity than tendons without pain. STUDY DESIGN: Cohort study. METHODS: 72 elite badminton players were interviewed about training, pain and injuries. The participants were scanned with high-end ultrasound equipment. Colour Doppler was used to examine the tendons of 64 players before a match and 46 players after a match. Intratendinous colour Doppler flow was measured as colour fraction (CF). The tendon complex was divided into three loci: the quadriceps tendon, the proximal patellar tendon and the insertion on the tibial tuberosity. RESULTS: Interview: Of the 72 players, 62 players had problems with 86 tendons in the lower extremity. Of these 86 tendons, 48 were the anterior knee tendons. Ultrasound: At baseline, the majority of players (87%) had colour Doppler flow in at least one scanning position. After a match, the percentage of the knee complexes involved did not change. CF increased significantly in the dominant leg at the tibial tuberosity; single players had a significantly higher CF after a match at the tibial tuberosity and in the patellar tendon both before and after a match. Painful tendons had the highest colour Doppler activity. CONCLUSIONS: Most elite badminton players had pain in the anterior knee tendons and intratendinous Doppler activity both before and after match. High levels of Doppler activity were associated with self-reported ongoing pain. PMID- 18381825 TI - Travelling to China for the Beijing 2008 Olympic Games. AB - The Beijing Olympics will be the world's premier sporting event in 2008. Countries should begin planning for appropriate support for their Olympic athletes. Experienced medical staff should be appointed early and become familiar with the major medical or injury concerns of athletes and officials. Teams should assemble a comprehensive medical kit. Appropriate medical advice can ensure that athletes will arrive in Beijing in optimal condition. Medical issues particularly relevant to competitors at the Beijing Olympics include heat and humidity, waterborne diseases and air pollution. However, the air pollution probably represents less of a threat than previous media reports would indicate. The Olympic village and venues can be thought of as small enclaves located within a rapidly developing country. PMID- 18381826 TI - The diagnosis of longstanding groin pain: a prospective clinical cohort study. AB - BACKGROUND: Longstanding groin pain is a difficult diagnostic challenge for sports physicians, and the lack of consensus on diagnostic criteria and taxonomy makes comparison of published studies difficult. AIM: To determine the usefulness and validity of the clinical classification proposed by Holmich et al in a primary care sports medicine population. DESIGN: Prospective cohort study. SETTING: Private sports medicine clinic in London, UK. PARTICIPANTS: 218 consecutive cases presenting with longstanding groin pain. INTERVENTIONS: Clinical assessment, diagnostic investigations and follow-up. RESULTS: Groin pain patients (173 men, 45 women) presented from 23 sporting codes from professional athletes to the recreational exerciser. Men most commonly played soccer (football) (22%) and rugby (21%), while women were most often runners (40%). 12 month follow-up was successful in 65% of cases; an accurate diagnosis was made in 89% of cases. Hip pathology (50.4%) was the most common form of injury, with pubic pathology seen in 21% of cases. Pubic pathology was most often seen in kicking sports (58%), and straight line activities most often resulted in hip pathology (39%). Those patients diagnosed with hip pathology were less likely to return to pre-morbid levels of activity than those diagnosed with pubic pathology (28% vs 15%). CONCLUSION: This series shows a different breakdown of injuries in a sporting population presenting with groin pain than previously reported. This reflects diagnostic difficulties in the area. The high incidence of hip pathology and the poor prognosis which this confers are worthy of note. PMID- 18381827 TI - Rethinking teaching nursing homes: potential for improving long-term care. AB - To meet the special needs of and provide quality health care to nursing home residents, the health care workforce must be knowledgeable about the aging process. Health professionals are minimally prepared in their academic programs to care for older adults, and few programs have required rotations in geriatrics. Teaching nursing homes (TNHs) have shown promise as sites for the preparation of a health workforce to care for older adults in nursing homes as well as improvement of quality outcomes. This article reports on the process and recommendations of a TNH summit of experts in geriatric education and practice as to the feasibility of developing a sustainable and replicable TNH model that would prepare a professional workforce knowledgeable about and prepared to work in long-term care. The TNH summit identified characteristics of partnerships between academia, nursing home(s), and other stakeholders that would constitute a successful TNH collaboration. Goals of a TNH partnership between service and academia include interdisciplinary education and practice, research and dissemination of evidence-based practices, and benchmarks of a nursing home professional learning environment. PMID- 18381828 TI - Globalization, women's migration, and the long-term-care workforce. AB - With the aging of the world's population comes the rising need for qualified direct long-term-care (DLTC) workers (i.e., those who provide personal care to frail and disabled older adults). Developed nations are increasingly turning to immigrant women to fill these needs. In this article, we examine the impact of three global trends-population aging, globalization, and women's migration-on the supply and demand for DLTC workers in the United States. Following an overview of these trends, we identify three areas with embedded social justice issues that are shaping the DLTC workforce in the United States, with a specific focus on immigrant workers in these settings. These include world poverty and economic inequalities, the feminization and colorization of labor (especially in long-term care), and empowerment and women's rights. We conclude with a discussion of the contradictory effects that both population aging and globalization have on immigrant women, source countries, and the long-term-care workforce in the United States. We raise a number of policy, practice, and research implications and questions. For policy makers and long-term-care administrators in receiver nations such as the United States, the meeting of DLTC worker needs with immigrants may result in greater access to needed employees but also in the continued devaluation of eldercare as a profession. Source (supply) nations must balance the real and potential economic benefits of remittances from women who migrate for labor with the negative consequences of disrupting family care traditions and draining the long-term-care workforce of those countries. PMID- 18381829 TI - Characterizing hospice services in the United States. AB - PURPOSE: Although caregivers desire specific information about hospice programs, there is little descriptive information available. We characterized agencies that provide formal or informal hospice care in the United States according to four types of services considered important by caregivers: medications and treatments; rehabilitative care; emotional, social, and spiritual support; and practical support (e.g., continuous home care). DESIGN AND METHODS: Data were from the nationally representative 2000 National Home and Hospice Care Survey. We categorized agencies into service mixes reflecting combinations of the four service types by using a grade-of-membership model to score each agency. RESULTS: Of the 11,419 agencies represented in our data, slightly more than half (52%) reflected some mix of services from all four service types. The remaining agencies provided service mixes that reflected relatively few or no services from at least one of the four service types. Specifically, approximately 7% lacked (i.e., had relatively few or no) medications and treatment services, 6% lacked rehabilitative care, 26% lacked emotional/social/spiritual support, and about 10% lacked multiple services. When compared to agencies that reflected a mix of all four services, agencies that lacked multiple services could be distinguished by their lack of formal certification as either a hospice or home health agency. IMPLICATIONS: Few U.S. agencies that provide formal or informal hospice care incorporate a balanced mix of all types of services, but most agencies lack only one type of service. Categorization of agencies based on services that are relevant to caregivers may assist providers in describing hospice care. PMID- 18381830 TI - Racial differences in hospice use and in-hospital death among Medicare and Medicaid dual-eligible nursing home residents. AB - PURPOSE: We investigated the role of race in predicting the likelihood of using hospice and dying in a hosptial among dual-eligible (Medicare and Medicaid) nursing home residents. DESIGN AND METHODS: This follow-back cohort study examined factors associated with hospice use and in-hospital death among non Hispanic Black and non-Hispanic White dual-eligible nursing home residents (N = 30,765) who died in Florida during one of three years: 2000, 2001, or 2002. We used logistic regression models to identify independent predictors of hospice use and in-hospital death. RESULTS: After we controlled for other factors, Black residents were significantly less likely to use hospice and more likely to die in a hospital. Principal cause of death moderated the relationship between race and hospice use: Black residents were significantly less likely to use hospice than White residents among residents without cancer as principal cause of death, but there was no difference among residents with cancer as cause of death. Further analyses for each racial group revealed that the impact of cause of death in predicting hospice use was greater among Black residents than White residents. IMPLICATIONS: Hospice care offers many benefits, including reduced risk of in hospital death, but Black nursing home residents are less likely to use hospice and may have different perceptions of need for hospice care compared with White residents. Future research and outreach efforts should focus on developing culturally sensitive, disease-focused end-of-life education and communication interventions that target residents, families, nursing home providers, and physicians. PMID- 18381831 TI - Welfare reform and older immigrants: food stamp program participation and food insecurity. AB - PURPOSE: The welfare reform bill of 1996 severely constrained noncitizens' eligibility for the Food Stamp Program (FSP). This study examined the effects of eligibility restrictions on older immigrants' FSP participation and food insecurity. We paid special attention to household composition and household eligibility as well as older immigrants' individual eligibility. DESIGN AND METHODS: The sample consisted of 3,175 low-income older adults from the 1999 Current Population Survey. We used probit and tobit regressions. We compared three analytical models: two models using the differences-in-differences approach with two different measures of immigration status (individual and household status), and one using a summary variable of the percentage of FSP-eligible people in a household. RESULTS: Both household immigration status and older adults' individual immigration status were significantly associated with the probability of FSP participation. Living in a household with a higher percentage of eligible members significantly increased older people's FSP participation and benefit levels while significantly reducing their risk of food insecurity. IMPLICATIONS: Noncitizen eligibility restriction affects older immigrants' FSP participation and food insecurity by its relation to household composition. Experts should therefore consider household composition when developing social policies for older immigrants. PMID- 18381832 TI - Self-efficacy and participation in physical and social activity among older adults in Spain and the United States. AB - PURPOSE: We explored Bandura's self-efficacy theory as applied to older adult (aged 63-92) participation in physical and social activity in a cross-cultural study. DESIGN AND METHODS: Older adults in Spain (n = 53) and the United States (n = 55) completed questions regarding self-efficacy, outcome expectancy, and participation in physical and social activities. RESULTS: Self-efficacy significantly predicted both physical and social activity in both Spain and the United States. Outcome expectancy did not significantly predict either activity, nor did education, gender, or overall health. Modified and new self-efficacy measures proved reliable in both samples. IMPLICATIONS: This study enhances understanding of how self-efficacy motivates participation in physical activity, as noted in previous studies, as well as provides a new understanding of what motivates participation in social activities. The high reliability of the new measures used in this study provides evidence for further use of these measures in other contexts. It is important to note that this study further supports the use of Bandura's theory of self-efficacy for cross-cultural applications. PMID- 18381833 TI - Volunteering and psychological well-being among young-old adults: how much is too much? AB - PURPOSE: Research concerned with the relationship between volunteer activity and psychological well-being has typically reported higher levels of well-being among older adult volunteers relative to nonvolunteers. However, few studies have examined nonlinear associations between frequency of volunteer activity and well being. We examined nonlinear associations between hours spent volunteering and psychological well-being, controlling for employment status, partner status, physical health, and education. We also investigated associations between different domains of volunteer activity and well-being, along with the possible moderating effects of gender on these relationships. DESIGN AND METHODS: We used data from the PATH Through Life Project, a population-based study of Australian adults. Participants consisted of 2,136 older adults aged 64 to 68. RESULTS: Nonlinear associations between hours spent volunteering and psychological well being were evident, with these associations characterized by inverted U shapes, with nonvolunteers and those volunteering at high levels producing lower well being scores relative to those volunteering at moderate levels. Few associations between specific domains of volunteer activity and well-being were evident, and no notable gender interactions emerged. IMPLICATIONS: The results point toward optimal frequency of engagement in volunteer activity for psychological well being as being bounded by upper and lower levels, outside of which benefits to well-being diminish. PMID- 18381834 TI - Racial differences in suicidality in an older urban population. AB - PURPOSE: This study used epidemiological data of older African Americans and Caucasians living in an urban community to compare those factors associated with active or passive suicidal ideation in each racial group. DESIGN AND METHODS: Using 1990 census data for Brooklyn, New York, we attempted to interview all cognitively intact adults aged 55 or older in randomly selected block groups. The sample consisted of 214 Whites and 860 Blacks. We adapted George's social antecedent model to examine 19 independent variables; the dependent variable was based on lifetime history of passive or active suicidal ideation (hereafter, suicidality). We weighted the sample by race and gender. To control for sampling design effects, we used SUDAAN for data analysis. RESULTS: Whites reported higher prevalence than Blacks for current suicidality (5.8% vs 2.3%) and lifetime suicidality (14.8% vs 10.2%). None of the differences were significant. In logistic regression analysis conducted for each race, four variables were associated with suicidality within both races: higher depressive symptom scores, higher anxiety symptom scores, copes by using medications, and lower religiosity. Two variables were associated with suicidality only among Whites: higher use of spiritualists and copes by keeping calm. One variable, greater use of doctors for mental health problems, was significant only among Blacks. IMPLICATIONS: There were no racial differences in the prevalence of suicidality. Virtually all of the factors associated with suicidality are potentially ameliorable. Among both racial groups, suicidality is likely to be impacted by addressing depressive and anxiety symptoms and, when appropriate, by encouraging various coping strategies, especially religiosity. PMID- 18381835 TI - Building quality report cards for geriatric care in The Netherlands: using concept mapping to identify the appropriate "building blocks" from the consumer's perspective. AB - PURPOSE: This article reports on a study to identify "building blocks" for quality report cards for geriatric care. Its aim is to present (a) the results of the study and (b) the innovative step-by-step approach that was developed to arrive at these results. DESIGN AND METHODS: We used Concept Mapping/Structured Conceptualization to define the building blocks. Applied to this study, we carried out Concept Mapping using several data collection methods: (a) a Web search, (b) semistructured interviews, (c) document analysis, (d) questionnaires, and (e) focus groups. RESULTS: The findings showed that, although home care and institutional care for elderly adults share many quality themes, experts need to develop separate quality report cards for the two types of geriatric care. Home care consumers attach more value to the availability, continuity, and reliability of care, whereas consumers of institutional care value privacy, respect, and autonomy most. This study also showed, unlike many other quality report card studies, that consumers want information on structure, process and outcome indicators, and rating outcome indicators such as effectiveness and safety of care both for home care and for institutional care. Concept Mapping proved to be a valuable method for developing quality report cards in health care. IMPLICATIONS: Building blocks were delivered for two quality report cards for geriatric care and will be used when quality report cards are built in The Netherlands. For the U.S. context, this study shows that current national report cards for geriatric care should be supplemented with quality-of-life data. PMID- 18381836 TI - Does mental illness affect consumer direction of community-based care? Lessons from the Arkansas Cash and Counseling program. AB - PURPOSE: Previous research from the Cash and Counseling Demonstration and Evaluation (CCDE) in Arkansas, New Jersey, and Florida suggests that giving consumers control over their personal care greatly increases their satisfaction and improves their outlook on life. Still, some argue that consumer-directed care may not be appropriate for consumers with intellectual disabilities or mental health diagnoses. This study examined how Cash and Counseling-a new option allowing consumers to manage an individualized budget equivalent to what agencies would have spent on their care-changes the way consumers with mental health diagnoses meet their personal care needs and how that affects their well-being. DESIGN AND METHODS: Using the Arkansas CCDE baseline and the 9-month follow-up data for individuals in the treatment and control groups, we compared and contrasted the experience of elderly consumers with and without mental health diagnoses utilizing logit regression. RESULTS: After examining several outcome measures, including satisfaction with care arrangements and the paid caregiver's reliability and schedule, unmet needs, and satisfaction with the relationship with paid caregivers, this study found evidence that, from the perspective of consumers, the Cash and Counseling program works well for participants with mental health diagnoses. IMPLICATIONS: Considering the growing need for long-term care services and the limited resources available, a consumer-directed option makes sense, and it can be a valuable alternative for persons with mental health needs. PMID- 18381837 TI - BE-ACTIV: a staff-assisted behavioral intervention for depression in nursing homes. AB - PURPOSE: This article (a) describes a 10-week, behavioral, activities-based intervention for depression that can be implemented in nursing homes collaboratively with nursing home activities staff and (b) presents data related to its development, feasibility, and preliminary outcomes. DESIGN AND METHODS: We developed BE-ACTIV, which stands for Behavioral Activities Intervention, in two pilot study phases: a treatment development phase and a feasibility-outcome phase with a small, randomized trial. We first piloted the intervention with five depressed residents in a single nursing home in collaboration with the social services and activities staff. The second phase randomized 20 residents from six nursing homes to receive either the intervention or treatment as usual. RESULTS: The intervention was well received by residents, family, and staff members. Experience with the intervention and input from staff members resulted in modifications to streamline the intervention and improve implementation. Results suggest that BE-ACTIV reduced institutional barriers to participation in pleasant activities, increased resident control over activity participation, increased overall activity participation, and improved depressive symptoms. Despite low power, statistical and graphical comparisons suggest superiority of the intervention over treatment as usual. IMPLICATIONS: Because depression among nursing home residents is prevalent, heterogeneous, and often treatment resistant, there is a need for effective, low-cost interventions that are ecologically acceptable and efficient. BE-ACTIV is a promising intervention; it is brief, addresses institutional barriers, involves facility staff in treatment, and is acceptable to residents. As such, BE-ACTIV merits further evaluation to establish efficacy and effectiveness. PMID- 18381838 TI - Assisted vacations for men with dementia and their caregiving spouses: evaluation of health-related effects. AB - PURPOSE: In this study, we conducted the first evaluation of assisted vacations for persons with dementia and their caregivers in the field of caregiving research. DESIGN AND METHODS: We used a quasi-experimental, two-group, repeated measures design with two measuring times (preintervention, 3-month follow-up) to examine whether assisted vacations lead to a reduction in physical complaints and symptoms of depression in family caregivers. The sample consisted of 29 caregiving wives who were providing at-home care for husbands suffering from dementia. We selected the participants in the control group from a waiting list. Outcome measurements were made with the Giessen Subjective Complaints List and the Beck Depression Inventory. RESULTS: The overall emotional and physical states of the participants in the intervention group showed significant improvements in comparison with those of the control group 3 months after the first interview. IMPLICATIONS: The results show that assisted vacations can have both immediate and longer lasting positive effects on participants' health. Assisted vacations can therefore be a way of diminishing the risk of stress disorders for caregiving spouses. Making assisted vacations available to people with dementia and their caregivers is a worthwhile goal for community support facilities working to reduce caregiver burden. PMID- 18381841 TI - Neurocognitive outcome following fetal growth restriction. AB - This review evaluates neurological and cognitive outcomes as they relate to fetal growth restriction. The aim is to clarify the relationship between poor fetal growth and abnormal brain development in order to understand the research directions required to untangle this complex issue. PMID- 18381840 TI - Liver gene expression analysis reveals endoplasmic reticulum stress and metabolic dysfunction in SCD1-deficient mice fed a very low-fat diet. AB - We previously reported that mice deficient in stearoyl-CoA desaturase-1 (Scd1) and maintained on a very low-fat (VLF) diet for 10 days developed severe loss of body weight, hypoglycemia, hypercholesterolemia, and many cholestasis-like phenotypes. To better understand the metabolic changes associated with these phenotypes, we performed microarray analysis of hepatic gene expression in chow- and VLF-fed female Scd1+/+ and Scd1-/- mice. We identified an extraordinary number of differentially expressed genes (>4,000 probe sets) in the VLF Scd1-/- relative to both VLF Scd1+/+ and chow Scd1-/- mice. Transcript levels were reduced for genes involved in detoxification and several facets of fatty acid metabolism including biosynthesis, elongation, desaturation, oxidation, transport, and ketogenesis. This pattern is attributable to the decreased mRNA abundance of several genes encoding key transcription factors, including LXRalpha, RXRalpha, FXR, PPARalpha, PGC-1beta, SREBP1c, ChREBP, CAR, DBP, TEF, and HLF. A robust induction of endoplasmic reticulum (ER) stress is indicated by enhanced splicing of XBP1, increased expression of the stress-induced transcription factors CHOP and ATF3, and elevated expression of several genes involved in the integrated stress and unfolded protein response pathways. The gene expression profile is also consistent with induction of an acute inflammatory response and macrophage recruitment. These results highlight the importance of monounsaturated fatty acid synthesis for maintaining metabolic homeostasis in the absence of sufficient dietary unsaturated fat and point to a novel cellular nutrient-sensing mechanism linking fatty acid availability and/or composition to the ER stress response. PMID- 18381839 TI - Effect of destrin mutations on the gene expression profile in vivo. AB - Remodeling of the actin cytoskeleton through actin dynamics (assembly and disassembly of filamentous actin) is known to be essential for numerous basic biological processes. In addition, recent studies have provided evidence that actin dynamics participate in the control of gene expression. A spontaneous mouse mutant, corneal disease 1 (corn1), is deficient for a regulator of actin dynamics, destrin (DSTN, also known as ADF), which causes epithelial hyperproliferation and neovascularization in the cornea. Dstn(corn1) mice exhibit an actin dynamics defect in the corneal epithelial cells, offering an in vivo model to investigate cellular mechanisms affected by the Dstn mutation and resultant actin dynamics abnormalities. To examine the effect of the Dstn(corn1) mutation on the gene expression profile, we performed a microarray analysis using the cornea from Dstn(corn1) and wild-type mice. A dramatic alteration of the gene expression profile was observed in the Dstn(corn1) cornea, with 1,226 annotated genes differentially expressed. Functional annotation of these genes revealed that the most significantly enriched functional categories are associated with actin and/or cytoskeleton. Among genes that belong to these categories, a considerable number of serum response factor target genes were found, indicating the possible existence of an actin-SRF pathway of transcriptional regulation in vivo. A comparative study using an allelic mutant strain with milder corneal phenotypes suggested that the level of filamentous actin may correlate with the level of gene expression changes. Our study shows that Dstn mutations and resultant actin dynamics abnormalities have a strong impact on the gene expression profile in vivo. PMID- 18381842 TI - Appropriate use of dopamine agonists and levodopa in restless legs syndrome in an ambulatory care setting. AB - BACKGROUND: Dopaminergic agents are the mainstay therapy for the management of restless legs syndrome (RLS). There are no clear guidelines on RLS management, and no study has evaluated the appropriate use of dopaminergic agents in RLS. OBJECTIVE: To evaluate the appropriateness of use of dopaminergic agents in RLS management in an ambulatory care setting based on the most current scientific evidence. METHODS: A retrospective drug utilization evaluation was conducted in patients who received levodopa or dopamine agonist for RLS from July 1, 2006, to July 31, 2007. Patients' medical records were reviewed and data were collected on demographics; comorbidities; laboratory values; doses of levodopa or dopamine agonists; prescribing physician's specialty; and use of alcohol, tobacco, and caffeine. RESULTS: A total of 27 patients were included in the study for data collection and analysis. Twenty-two (81%) patients were on levodopa and 5 (19%) were on ropinirole. RLS severity was documented in only 2 (7%) patients. Serum ferritin levels and transferrin-iron saturation (Tsat) percentages were not obtained in 18 (67%) and 20 (74%) of the patients, respectively. Two (7%) patients had ferritin levels less than 50 ng/mL, and 7 (26%) patients had ferritin levels greater than 50 ng/mL. Fourteen (52%) patients were taking concurrent antidepressants and 6 (22%) were taking sedating antihistamines. Alcohol and tobacco use was documented in 2 (7%) and 8 (30%) patients, respectively. Twenty-six (96%) of the prescribing physicians were primary care providers. CONCLUSIONS: The findings of this study confirm the need for provider education about the appropriate use of levodopa and dopamine agonists in patients with RLS. Appropriate use of these drugs may help decrease unnecessary adverse effects, complications, and costs. PMID- 18381843 TI - Very late stent thrombosis after dual antiplatelet therapy discontinuation in a patient with a history of acute stent thrombosis. AB - OBJECTIVE: To describe a case of very late stent thrombosis after dual antiplatelet discontinuation in a patient with a previous history of stent thrombosis. CASE SUMMARY: A 62-year-old man with a history of coronary artery disease, multiple acute coronary syndromes requiring percutaneous coronary interventions with multiple stent placements, and acute stent thrombosis resulting in ST segment elevation myocardial infarction presented to the hospital with chest pain. The chest pain was not relieved by 4 sublingual nitroglycerin tablets. Five days prior to his presentation, the patient had been instructed to discontinue both aspirin and clopidogrel in preparation for a left ankle fusion procedure. He was taken to the cardiac catheterization laboratory where he was found to have thrombosis in a sirolimus-eluting stent placed more than 3 years ago. Thrombectomy and balloon angioplasty were performed, and the patient completed his hospital course without complications. DISCUSSION: Stent thrombosis associated with drug-eluting stents is a complicated pathophysiologic phenomenon with multiple patient-, procedure-, and device-related factors. Application of these risk factors to quantify the risk of stent thrombosis as they apply to a single patient is unknown. Discontinuation of recommended dual antiplatelet therapy with aspirin plus a thienopyridine has been identified as a major risk factor for stent thrombosis, but the optimal duration of dual antiplatelet therapy remains unknown. Current recommendations suggest extending dual antiplatelet therapy beyond one year in patients with low bleeding risk. CONCLUSIONS: Given the overall data at this time and the severity of stent thrombosis, it seems prudent to continue dual antiplatelet therapy with aspirin indefinitely plus a thienopyridine for at least one year, with continuation beyond one year on a case-by-case basis depending on the risks of in-stent thrombosis and bleeding. In patients with a low risk of bleeding, indefinite continuation of dual antiplatelet therapy may be reasonable. PMID- 18381844 TI - Daptomycin-induced acute renal and hepatic toxicity without rhabdomyolysis. AB - OBJECTIVE: To report a case of a patient who experienced acute renal and hepatic toxicity following administration of daptomycin and review previously published case reports of renal and hepatic dysfunction with daptomycin. CASE SUMMARY: A 35 year-old man receiving daptomycin 4 mg/kg (275 mg) intravenously once daily (started 5 wk prior to presentation for presumed osteomyelitis) presented to the emergency department with elevations in serum creatinine and hepatic transaminase levels. He did not experience creatine kinase (CK) elevation or rhabdomyolysis. Following discontinuation of daptomycin, his renal and hepatic function improved. DISCUSSION: To our knowledge, this is the first case of daptomycin-induced hepatotoxicity with acute renal failure in the absence of rhabdomyolysis and CK abnormalities. Previously published case reports described patients with a variety of elevations in liver function tests, serum creatinine, and CK with daptomycin. In our patient, the acute renal and hepatic toxicity was probable according to the Naranjo probability scale. CONCLUSIONS: Although daptomycin is a well-tolerated antibacterial agent, clinicians should consider periodic monitoring of liver function and renal function tests to identify potential adverse effects. PMID- 18381845 TI - Surface replicas of normal and vitrified leaves of Datura insignis, Barb Rodr. AB - Vitrification is a morphological and physiological disorder affecting plants during their in vitro vegetative propagation. Vitrified plants have a poor survival rate when transferred from in vitro to greenhouse conditions, a fact mainly due to water loss and dissecation. It has been shown that normal and vitreous leaves of Datura insignis differ in the frequency of normal and abnormal stomata. The purpose of this work was to compare the surface of normal and vitreous leaves of D. insignis, using a modification of the platinum/carbon replica method. Adaxial and abaxial leaf surfaces of normal plantlets have a smooth and homogenous cuticle. A granular aspect, probably due to leaf age, rarely occurs at the periphery of the epidermal cells. Both adaxial and abaxial leaf surfaces of vitrified plantlets show discontinuities in the cuticle, occurring at several regions of the outer periclinal cell walls. However, such discontinuities are most noticeable in the region between adjacent epidermal cells. Fibrils 20-30 nm thick show a random arrangement or an oriented pattern in cuticular discontinuities. In D. insignis vitrified plantlets, adaxial and abaxial leaf cuticle has discontinuities or gaps which may cause an increase in cuticular transpiration contributing to the low survival rate of vitrified plantlets. PMID- 18381846 TI - Educational and career pathways of dental hygienists: comparing graduates of associate and baccalaureate degree programs. AB - The purpose of this study was to compare the educational and career pathways of graduates from associate degree (A.S./A.A.) and baccalaureate degree (B.S./B.A.) entry-level dental hygiene (DH) programs. A thirty-item, closed-ended questionnaire was mailed to a random sample of 1,352 dental hygienists who were educated and licensed in California between 1990 and 2000. The response rate was 76 percent. Approximately half of the respondents had a degree prior to entering the DH program, and the elapsed time between degrees was greater for A.S./A.A. graduates (p=0.05). More B.S./B.A. graduates had earned or were seeking master's and other more advanced degrees: 11.9 percent versus 4.3 percent. Graduates from both types of programs were involved in professional and community organizations and held leadership positions. Most were currently practicing in the traditional clinical setting, with means of 3.6 and 3.3 days/week for the A.S./A.A. and B.S./B.A. graduates, respectively (p<0.05). More B.S./B.A. graduates held DH faculty positions (30.3 percent versus 4.3 percent, p<0.05) and other non-DH teaching positions (14.9 percent versus 8.6 percent, p<0.05) and had greater involvement with research (8.0 percent versus 3.6 percent, p<0.05). The two groups did not differ in regard to other dental and DH-related positions. In conclusion, graduates from B.S./B.A. programs are more likely to have positions in nontraditional settings. PMID- 18381847 TI - Dental education in a flat world: advocating for increased global collaboration and standardization. AB - Globalization is a broad term referring to the increasing connectivity, integration, and interdependence of economies, societies, technologies, cultures, and political and ecological spheres across the world. This position paper was developed by a working group of the 2007 American Dental Education Association (ADEA) Leadership Institute. The authors explore the effect that globalization has had on dentistry and dental education to date and hypothesize what dental education could look like in the years ahead. While the paper is written from a North American perspective, some of the authors bring international expertise and experience to the topic of global dental education in a flat world. Specific issues and barriers addressed in this position paper include variations in accreditation and licensure requirements in dental education throughout the world; the historical development of dental education models (odontology and stomatology) and the need for congruency of these models in the global environment; the competency-based model of education and its relevance to development and implementation of global dental competencies; and the slow adoption of technological advances in dental education for promoting collaborations and encouraging resource sharing among countries. These challenges are discussed as they affect the implementation of a standardized global dental education that can lead to improved access to oral health care services and better oral and overall health for the citizens of the world. PMID- 18381848 TI - Dentists' comfort in treating underserved populations after participating in community-based clinical experiences as a student. AB - The purpose of this project was to determine new dentists' comfort levels in treating traditionally underserved populations after participating in two consecutive five-week community-based clinical experiences while in dental school. A written survey was mailed to all known University of Iowa alumni (1992 2002; N=745). Respondents were asked to rank their comfort levels in treating twelve underserved populations on a five-point Likert type scale (5=no problem; 1=will not). Bivariate and logistic regression model analyses were performed to examine associations (p<0.05) among comfort and six predictor variables. Alumni (n=372) were most comfortable treating other ethnic, low-income, non-English speaking, and HIV+/AIDS populations and least comfortable treating incarcerated and homebound populations. The following variables were significantly associated with comfort: 1) perception that the community experiences had great/much value; 2) practice located in larger communities; 3) non-solo practitioners; and 4) dentist's gender. As more dental schools utilize community-based clinical experiences to increase students' exposure to underserved populations, it is important that these experiences provide exposure to a variety of populations. Additionally, dental schools should continuously monitor the short- and long-term value of these programs for their students and recent graduates. PMID- 18381849 TI - Attitudes toward patient safety standards in U.S. dental schools: a pilot study. AB - The objective of this study was to assess the patient safety culture among students, staff, and faculty in seven U.S. dental school clinics when compared to those from a similar study in twenty U.S. hospitals. A survey on patient safety culture developed by the Agency for Healthcare Research and Quality (AHRQ) was used to measure attitudes towards patient safety by anonymous faculty, students, and support staff members who work in the clinics of seven U.S. dental schools. This survey instrument was also administered to staff at twenty U.S. hospitals. In three of the twelve sections of the survey (Overall Perceptions of Safety, Management Support for Patient Safety, and Teamwork Across Units), dental school personnel responses rated above the hospital benchmark results. In Section 2 (Frequency of Adverse Events Reported) and Section 4 (Organizational Learning/ Continuous Improvement), average dental school responses were below those recorded for hospital personnel. The overall score from the twelve sections of the survey indicated that patient safety attitudes of dental school participants were higher than those of their hospital counterparts. PMID- 18381850 TI - The Academic Dental Careers Fellowship Program: a pilot program to introduce dental students to academia. AB - The Academic Dental Careers Fellowship Program (ADCFP) was established in 2006 by the American Dental Education Association (ADEA) and the American Association for Dental Research (AADR) with the financial support of the ADA Foundation to encourage dental students to consider careers in dental education and to provide participating fellows with insights into academic life. The ADA Foundation provided funding during the 2006-07 academic year for eleven dental student fellows, who were paired with faculty mentors at their respective schools. Fellows and mentors attended a two-day retreat in the summer of 2006, and over the course of the subsequent year in dental school, the fellows with guidance from their mentors participated in preclinical laboratory, classroom, small group, and clinical teaching experiences; designed and implemented a research project; developed a philosophy of education; completed career reflection essays; assembled a portfolio to represent their ADCFP activities and projects; conducted a series of interviews with faculty designed to expose students to roles, issues, and career paths in academic dentistry; and presented a synopsis of their experiences at the ADEA Annual Session in New Orleans in March 2007. The fellows and mentors completed midyear and end-of-year evaluations of the ADCFP in which feedback and recommendations were collected by telephone interviews and questionnaires. Fellows reported positive experiences and an increased interest in and understanding of academic careers. Mentors also evaluated the ADCFP positively and reported enhancements in their mentoring skills. This article describes the goals and format of the ADCFP, summarizes program evaluation data elicited from the fellows and mentors, and proposes recommendations for future fellowship classes. PMID- 18381851 TI - Reflections on academic careers by current dental school faculty. AB - During the inaugural year (2006-07) of the Academic Dental Careers Fellowship Program (ADCFP), 110 faculty members at ten different dental schools were interviewed by dental students who were participating as ADCFP fellows in this year-long program designed to introduce them to faculty roles and activities and help them gain an appreciation for the rewards and issues associated with academic life. The goals, format, and components of the ADCFP are described in a companion article in this issue of the Journal of Dental Education. One of the fellows' assignments during the ADCFP was to interview faculty at various academic ranks who had differing degrees of work emphasis in teaching, research, service/patient care, and administration. Sixty-nine (63 percent of the total) of these interviews were reviewed and analyzed by the authors, who were student fellows in the ADCFP during 2006-07. The purpose of these interviews was to provide the fellows with insight into the positive aspects and challenges in becoming and remaining a dental school faculty member. This aggregate perspective of the interviews conducted at ten dental schools highlights the motivations and challenges that confront a dentist during the process of choosing a career in academic dentistry and determining if dental education is a good fit for each individual who elects to pursue this pathway. Thematic analysis of the interviews revealed several factors consistently identified by faculty across the schools as being positive influences on the quality of the academic work environment and career satisfaction: mentorship and student interaction, opportunities for scholarship (research and discovery), job diversity, intellectual challenge, satisfaction with the nature of academic work, lifestyle/family compatibility, flexibility, lifelong learning, professional duty, and lab responsibility. A series of negative themes were also consistently identified: bureaucracy/administrative burdens and barriers, time commitment, financial frustration, political frustration, lack of mentorship, required research emphasis, lack of teaching skills development, student engagement, isolation, and funding uncertainty. This article reports the approximate frequency of each theme, presents representative statements that describe the motivations and attitudes of dental faculty members who were interviewed, and concludes with a review of programs/methods aimed at marketing academic careers to current students. The purpose of this review of the rewards, benefits, and challenges that current dental faculty face is to provide students who are considering dental education with a frame of reference to guide their further exploration of this career path and to help students appreciate the many positive aspects of academic life that may not be readily apparent from their own interactions with faculty members. PMID- 18381853 TI - Faculty perceptions of a professional development seminar. AB - A web-enhanced seminar was provided to address the pedagogical needs of professional faculty educators at a college of dentistry in a research-intensive university in fall 2006. In this qualitative report, the authors describe the participants' (n=12) perceptions and evaluation of the seminar. An evaluation of the seminar journals showed documented improvement in the participants' confidence and an enhanced awareness of their teaching practices. Six themes (new knowledge, planned change, awareness, changes made, current practice, and challenges to learning) ranging from 5.3 percent to 35.5 percent among four to twelve participants emerged across their learning journals. Participants also rated the course 4.9 on a five-point scale in helping them understand a variety of teaching modalities other than lecture and the sole use of multiple-choice tests as insufficient. When we invest resources in our faculty, progress is made towards ensuring quality teaching, as well as increased understanding and enhanced communication in the teaching and learning environment. This recursive process not only influences the faculty and those they teach but the patients they care for as well. PMID- 18381852 TI - The state of the art in evaluating the performance of assistant and associate deans as seen by deans and assistant and associate deans. AB - This study explores the little-understood process of evaluating the performance of assistant and associate deans at dental colleges in the United States and Canada. Specifically, this research aimed to identify the methods, processes, and outcomes related to the performance appraisals of assistant/associate deans. Both deans and assistant/associate deans were surveyed. Forty-four of sixty-six deans (66.7 percent) and 227 of 315 assistant/associate deans (72.1 percent) completed surveys with both close-ended and open-ended questions. In addition, ten individuals from each group were interviewed. Results indicate that 75-89 percent of assistant/associate deans are formally evaluated, although as many as 27 percent may lack formal job descriptions. Some recommended best practices for performance appraisal are being used in a majority of colleges. Examples of these best practices are having at least yearly appraisals, holding face-to-face meetings, and setting specific, personal performance objectives/benchmarks for assistant/associate deans. Still, there is much room to improve appraisals by incorporating other recommended practices. Relatively high levels of overall satisfaction were reported by both assistant/associate deans and deans for the process and outcomes of appraisals. Assistant/associate deans rated the value of appraisals to overall development lower than did deans. Qualitative data revealed definite opinions about what constitutes effective and ineffective appraisals, including the use of goal-setting, timeliness, and necessary commitment. Several critical issues related to the results are discussed: differences in perspectives on performance reviews, the importance of informal feedback and job descriptions, the influence of an assistant/associate deans' lack of tenure, and the length of service of deans. Lastly, recommendations for enhancing performance evaluations are offered. PMID- 18381854 TI - Characteristics of effective classroom teachers as identified by students and professionals: a qualitative study. AB - This qualitative research study identified criteria for teacher quality preferences as perceived by current and past students. A two-question, open-ended survey asking what qualities learners liked most and least in a teacher/presenter was given to two groups: students (Group A) from medicine, dentistry, and related residency programs; and dentists and physicians (Group B) who had graduated at least three years previously and who attended a minimum of two days of continuing education courses in lecture format each year. A total of 300 subjects provided 2,295 written responses. Descriptive words within the responses were coded and grouped according to similar relationships, resulting in the emergence of twenty one defined categories that were further refined into three core categories: personality, process, and performance. Results showed that the two groups appear to have different preferences in teacher/presenter characteristics. For Group A (students), the categories of content design, content organization, and content development were at the forefront of their preferences. Group B (professionals) overwhelmingly favored elements of speaker self-confidence and expertise. Both groups highly valued expertise and speaking style. These findings can be used to develop curriculum, enhance faculty members' teaching skills, and plan continuing education programs. PMID- 18381855 TI - A virtual system for cavity preparation in endodontics. AB - This article presents a novel virtual teeth drilling system designed to aid dentists, dental students, and researchers in getting acquainted with teeth anatomy, the handling of drilling instruments, and the challenges associated with drilling procedures during endodontic therapy. The system is designed to be used for educational and research purposes in dental schools. The application features a 3D face and oral cavity model constructed using anatomical data that can be adapted to the characteristics of a specific patient using either facial photographs or 3D data. Animation of the models is also feasible. Virtual drilling using a Phantom Desktop (Sensable Technologies Inc., Woburn, MA) force feedback haptic device is performed within the oral cavity on 3D volumetric and surface models of teeth, obtained from serial cross sections of natural teeth. Final results and intermediate steps of the drilling procedure can be saved on a file for future use. The application has the potential to be a very promising educational and research tool that allows the user to practice virtual teeth drilling for endodontic cavity preparation or other related procedures on high detail teeth models placed within an adaptable and animated 3D face and oral cavity model. PMID- 18381856 TI - Low-frequency protamine 1 gene transversions c.102G->T and c.-107G->C do not correlate with male infertility. PMID- 18381858 TI - Intimal sarcoma of aortic arch treated with proton therapy following surgery. AB - Management of a rare case of intimal sarcoma of the aortic arch is reported, which was diagnosed unexpectedly after total arch replacement for pseudoaneurysm. The prognosis for this condition is poor, with death usually within a few months from diagnosis. The newly developed proton-beam radiation therapy was applied to treat a local recurrence of the sarcoma following surgery. Positron-emission tomography/computed tomography revealed complete remission of the lesion. PMID- 18381859 TI - Mycotic pseudoaneurysm of the ascending aorta at site of aortic cannulation. AB - Reoperation for pseudoaneurysm of the ascending aorta presents a surgical challenge. Instituting femorofemoral bypass and establishing hypothermic circulatory arrest is a well-known strategy, although not free from complications. We report a case of mycotic pseudoaneurysm after coronary artery bypass in a 53-year-old man, at the site of previous aortic cannulation, and review the surgical strategies proposed to manage this pathologic entity. PMID- 18381860 TI - Double outlet right ventricle with anomalous left pulmonary artery. AB - We describe a rare case of anomalous origin of the left pulmonary artery from the ascending aorta with concomitant double-outlet right ventricle in a 2-year-old boy. He underwent successful 2-stage surgical treatment with transluminal balloon pulmonary valvuloplasty, followed by complete repair. A follow-up examination at 4 years after the operation showed good results. PMID- 18381861 TI - Chylous pericardial effusion after aortic valve replacement. AB - Chylous pericardial effusion after open-heart surgery is a rare complication. We report a case of chylous pericardial effusion following aortic valve replacement, which presented as cardiac tamponade, and its subsequent management. PMID- 18381862 TI - From vision to mission in myocardial restoration. PMID- 18381863 TI - Procedural outcome and midterm result of carotid stenting in high-risk patients. AB - Carotid endarterectomy is the standard treatment for carotid stenosis, but carotid artery stenting has emerged as a potential alternative. Elective carotid artery stenting was performed in 42 patients aged 42 to 79 years (mean, 67.05 +/- 8.67 years) after ultrasonography, computed tomography, magnetic resonance angiography and a neurological evaluation. There was bilateral carotid stenosis in 23 patients (55%), with > 90% stenosis in 18 vessels. All patients had significant associated coronary lesions. An emboli protection device and self expanding stents were used. One year later, the patients were evaluated by Doppler sonography and selective angiography. Technical success was achieved in all procedures. During follow-up, 1 (2.4%) patient died from myocardial infarction, 1 underwent coronary artery bypass and 14 (40%) had minor complaints including occasional dizziness. No other neurological events were noted. Restenosis was found in one case, but selective angiography ruled out a significant lesion. One patient suffered embolization, but recovered completely within 24 hours. In 7 (17%) patients with type C arch interruption and a tortuous carotid course, stenting was successful and they had no embolization or restenosis. Carotid artery stenting is recommended in high-risk patients. PMID- 18381864 TI - Cardiac troponin I vs EuroSCORE: myocardial infarction and hospital mortality. AB - Perioperative myocardial infarction is the most common cause of morbidity and mortality in cardiac surgery. It occurs in 8% to 35% of patients. The primary aim of this prospective study was to determine the level of cardiac troponin I that indicates perioperative myocardial infarction in patients undergoing coronary artery bypass. A secondary goal was to establish the best independent predictor of hospital death. There were 180 consecutive patients undergoing isolated coronary artery bypass surgery enrolled in this study. Values of cardiac troponin I > 12.9 ng.mL(-1) at 8 hours postoperatively predicted perioperative myocardial infarction with a sensitivity of 100% and a specificity of 93.2%. Compared to patients who survived, those who suffered hospital death were significantly older (74 +/- 7 vs 63 +/- 10 years), had significantly higher levels of cardiac troponin I at 24 hours (9 +/- 17 vs 27.3 +/- 16 ng.mL(-1)) and 48 hours (6.9 +/- 19 vs 30.3 +/- 24 ng.mL(-1)) postoperatively, and a significantly higher EuroSCORE (9 +/- 2 vs 4 +/- 3). At 8 hours postoperatively, cardiac troponin I led to an earlier diagnosis of perioperative myocardial infarction, while EuroSCORE was the strongest independent predictor of hospital death. PMID- 18381865 TI - Differentiation of ischemic and dilated cardiomyopathy on electrocardiograms. AB - Differentiating coronary artery disease with left ventricular dysfunction from dilated cardiomyopathy is important prognostically and therapeutically. To provide a diagnostic algorithm to distinguish these conditions using a standard 12-lead electrocardiogram, all 105 patients with left ventricular ejection fraction < 50% who underwent angiography between January 2004 and December 2006 were studied prospectively. Coronary artery disease was defined as >/= 50% stenosis of the left main coronary artery or >/= 70% stenosis of 1 or more of the 3 major epicardial arteries. Normal coronary angiography findings with left ventricular ejection fraction < 50% was defined as dilated cardiomyopathy. The most specific finding for differentiation of these diseases was pathologic Q waves in lead II, aVF, V3 or V4. The most sensitive parameter was a ratio >/= 5 of R-wave amplitudes in lead V6 and lead III (94% sensitive). The 12-lead electrocardiogram provides a useful noninvasive method for differentiation of dilated cardiomyopathy from coronary artery disease with left ventricular systolic dysfunction. PMID- 18381866 TI - Valvuloplasty in the treatment of rheumatic tricuspid disease. AB - Organic involvement of the tricuspid valve is uncommon. Valve repair is preferred over replacement as it results in a low gradient across the valve and obviates the risk of prosthesis-related complications. From October 2002 to October 2004, 37 patients who required tricuspid valve repair for organic involvement were included in this study. They were divided into 2 groups depending on the surgical procedure for valve repair: 20 patients in group 1 had tricuspid commissurotomy and De Vega annuloplasty; 17 in group 2 had tricuspid commissurotomy only. There were significant reductions in peak and mean tricuspid gradients and right ventricular systolic pressure in both groups. Annular shortening was similar in both groups (median, 23% in group 1, 21% in group 2), but the ratio of the tricuspid regurgitation jet area to right atrial area was greater in group 2 (median, 0.40 in group 2, 0.19 in group 1). There was no postoperative death. We recommend supporting the tricuspid annulus with annuloplasty in patients with organic tricuspid valve disease and no dilatation of the annulus, if annular shortening is < 30%. PMID- 18381867 TI - Abscess of residual lobe after pulmonary resection for lung cancer. AB - Abscess of the residual lobe after lobectomy is a rare but potentially lethal complication. Between January 1975 and December 2006, 1,460 patients underwent elective pulmonary lobectomy for non-small-cell lung cancer at our institution. Abscess of the residual lung parenchyma occurred in 5 (0.3%) cases (4 bilobectomies and 1 lobectomy). Postoperative chest radiography showed incomplete expansion and consolidation of residual lung parenchyma. Flexible bronchoscopy revealed persistent bronchial occlusion from purulent secretions and/or bronchial collapse. Computed tomography in 3 patients demonstrated lung abscess foci. Surgical treatment included completion right pneumonectomy in 3 patients and a middle lobectomy in one. Complications after repeat thoracotomy comprised contralateral pneumonia and sepsis in 1 patient. Residual lobar abscess after lobectomy should be suspected in patients presenting with fever, leukocytosis, bronchial obstruction and lung consolidation despite antibiotic therapy, physiotherapy and bronchoscopy. Computed tomography is mandatory for early diagnosis. Surgical resection of the affected lobe is recommended. PMID- 18381868 TI - Survival after surgery with cardiopulmonary bypass in low weight patients. AB - To evaluate risk factors for hospital death in patients weighing < 2.5 kg undergoing open-heart surgery, records of 34 consecutive low-weight patients operated on between December 1997 and November 2004 were reviewed. Mean weight was 2.152 +/- 0.237 kg (range, 1.600 to 2.460 kg). Biventricular repair was achieved in 28 patients. The most frequent procedures were the arterial switch operation in 9 children, ventricular septal defect closure in 6, repair of total anomalous pulmonary venous connection in 5 and truncus arteriosus repair in 5. There were 8 early deaths. Mortality was strongly associated with the Comprehensive Aristotle Complexity Score: mortality was low (2/27; 7.4%) with a score < 19, and high (6/7; 85.7%) with a score >/= 19. Higher mortality was encountered after univentricular repair (4/6; 67%). Hyperlactatemia at the end of cardiopulmonary bypass was also associated with poor survival. A Comprehensive Aristotle score < 19 was the strongest predictor of survival in low-weight patients undergoing open-heart surgery. Biventricular repair, when feasible, should be promoted to improve outcome. PMID- 18381869 TI - Management of malignant pleural effusion associated with trapped lung syndrome. AB - Management of recurrent malignant pleural effusion associated with trapped lung syndrome remains problematic. An alternative treatment using a pleural catheter has been advocated. Between August 1999 and August 2002, 127 patients underwent thoracoscopy for malignant pleural effusion. Of these, 52 (41%) with trapped lung were managed by insertion of a pleural catheter. Mean age was 66 years (range, 42 89 years). The most frequent diagnosis was breast cancer. Spontaneous pleurodesis (drainage < 10 mL) occurred in 25 (48%) patients whose catheter was removed after 30 to 255 days (mean, 93.8 days). Symptomatic relief was achieved in 49 (94%) patients. Mean dyspnea score improved significantly from 3.0 to 1.9. Complications comprised catheter blockage, surgical emphysema, cellulitis, and loculated effusion in 2 patients each. Mean length of hospital stay was 3 days (range, 1-16 days). Median survival was 126 days (range, 10-175 days). We conclude that long-term placement of a pleural catheter provides effective palliation for malignant pleural effusion associated with trapped lung syndrome. PMID- 18381870 TI - Potential role of adipocytokine leptin in acute coronary syndrome. AB - By activating immune cells or a direct action on the vascular wall, leptin may affect the initiation and progression of atherosclerosis. We investigated whether plasma leptin concentration is associated with coronary artery disease, with particular focus on the relationship between plasma leptin and the development of an acute coronary syndrome. Plasma leptin, interleukin-6 and high-sensitivity C reactive protein were measured in 34 patients with acute coronary syndrome and 21 with stable angina. Their results were compared with those of 21 normal controls. Plasma leptin levels were significantly higher in the acute coronary syndrome group (13.36 +/- 5.02 ng.mL(-1)) compared to the stable angina group (8.97 +/- 4.06 ng.mL(-1)) or normal controls (5.14 +/- 2.75 ng.mL(-1)). Interleukin-6 and high-sensitivity C-reactive protein were also higher in the acute coronary syndrome group, and leptin correlated positively with interleukin-6 and high sensitivity C-reactive protein. These findings suggest that plasma leptin levels may be a useful marker of systemic inflammation, and measurement of plasma leptin may be helpful in assessing the risk of developing coronary heart disease. PMID- 18381871 TI - Valve repair in rheumatic heart disease in pediatric age group. AB - Valve repair in children is technically demanding but more desirable than valve replacement. From April 2004 to September 2005, 1 boy and 8 girls with rheumatic heart disease, aged 2-13 years (median, 9 years), underwent valve repair for isolated mitral regurgitation in 5, combined mitral and aortic regurgitation in 2, mitral stenosis in 1, and mitral regurgitation associated with atrial septal defect in 1. Chordal shortening in 7, annular plication in 6, commissurotomy in 1, reconstruction of commissural leaflets in 7 were performed for mitral valve disease. Plication and reattachment of the aortic cusps was carried out in 2 patients. Annuloplasty rings were not used. All patients survived the operation, 8 had trivial or mild residual mitral regurgitation, and 1 had trivial aortic regurgitation. Mean left atrial pressure decreased from 14 to 7 mm Hg postoperatively. During follow-up of 3-18 months, all children were asymptomatic and enjoyed normal activity. None required reoperation. In addition to chordal shortening and annular plication, reconstruction of the commissural leaflets is considered the most important aspect of valve repair. It can be achieved without annuloplasty rings, giving good early and midterm results. PMID- 18381872 TI - Endoscopic vs conventional vein harvesting: a prospective analysis. AB - Minimally invasive vein harvesting is associated with better leg wound healing and a lower incidence of wound infections. We analyzed our experience in 2 prospectively enrolled groups of non-randomized patients undergoing elective coronary artery bypass grafting. Group 1 was 81 patients who had endoscopic vein harvesting; group 2 was 80 who had conventional open vein harvesting. The time taken for endoscopic harvest (skin incision to skin closure) was significantly less than that for open harvest (51.07 vs 75.94 min). The number of cases to reach a plateau on the learning curve for endoscopic vein harvest was 20 for 2 lengths of vein and 35 for 3 lengths of vein. Significantly more suture repairs per vein were required in group 1 (1.32) than group 2 (0.38). The incidence of wound infection was 1.2% in group 1 vs 8.8% in group 2. Endoscopic vein harvesting is not difficult to learn and it should be preferred over open vein harvest, given its benefits in wound healing. PMID- 18381873 TI - Solitary benign fibrous tumors of the pleura. AB - Solitary benign fibrous tumors of the pleura are very rare. Seven patients (mean age, 52 years) who presented between 1995 and 2005 were studied retrospectively. They had nonspecific chest symptoms and no history of asbestos exposure or smoking. Chest radiography showed a large opacity occupying most of the affected hemithorax, with clear costophrenic angles. Computed tomography of the chest showed a large well-delineated heterogeneous mass directly related to the lateral chest wall. Needle biopsy suggested the benign nature of the lesion. Surgery was performed through a posterolateral thoracotomy. Five tumors arose from visceral pleura, and 2 from parietal pleura. The mean tumor diameter was 7.5 cm (range, 8 14 cm). Complete surgical excision was carried out in all cases. Histopathology and immunohistochemical staining confirmed the benign nature of the tumors. There was no mortality or major complication. The mean follow-up period was 4 years (range, 1-10 years). All patients remained tumor-free during follow-up. Wide local excision, including pulmonary and pleural resection, is recommended as the best therapeutic option. PMID- 18381874 TI - Intramyocardial angiogenic cell precursor injection for cardiomyopathy. AB - Stem cell therapy for heart failure is a rapidly progressing field. The objective of this study was to assess the safety, and short-term results of thoracoscopic direct injection of angiogenic cell precursors into patients with endstage cardiomyopathy. Cells were obtained from the patient's own blood, avoiding immunological concerns. The number of cells prior to injection was 29.1 +/- 18.9 x10(6). Forty-one patients with cardiomyopathy (mean age, 58.5 +/- 14.3 years) underwent stem cell injection; 21 had dilated cardiomyopathy and 20 had ischemic cardiomyopathy. Overall ejection fraction improved significantly by 4.8% +/- 7.5% at 149 +/- 98 days postoperatively. It increased from 25.9% +/- 8.6% to 28.7% +/- 9.8% in dilated cardiomyopathy, and from 26.6% +/- 5.8% to 33.6% +/- 7.8% in ischemic cardiomyopathy. New York Heart Association functional class was significantly better at 2 months in both groups. It was concluded that thoracoscopic intramyocardial angiogenic cell precursor injection is feasible and safe in patients with cardiomyopathy. The early results are good, and phase II trials are in progress. PMID- 18381875 TI - Late presentation of cor triatriatum with persistent levoatrial cardinal vein. AB - An asymptomatic 10-year-old boy presented with reduced exercise tolerance and an echocardiographic diagnosis of cor triatriatum. Transthoracic and transesophageal echocardiography failed to reveal the persistent levoatrial cardinal vein discovered at surgery. In patients with late presentation of cor triatriatum with severe mitral inflow obstruction and a small patent foramen ovale, an alternative communication between the posterior collecting chamber and the systemic venous circulation should be sought with alternative imaging techniques. PMID- 18381876 TI - Fibrin sealant for left ventricular rupture after mitral valve replacement. AB - Despite safer surgical procedures, left ventricular rupture remains a rare but potentially lethal complication of mitral valve replacement. The use of fibrin sealant has substantially improved the outcome of many difficult bleeding episodes after cardiac surgery. We describe a case of left ventricular rupture successfully treated with fibrin sealant combined with external Teflon-pledgeted sutures. PMID- 18381877 TI - Angiosarcoma presenting as syncope. AB - A 31-year-old lady presented with anemia and syncope. Echocardiography revealed massive pericardial effusion with a right atrial mass. Transesophageal echocardiography, computed tomography and magnetic resonance imaging scans confirmed presence of a right atrial mass. Histopathology revealed a high grade angiosarcoma. Complete resection was done and the patient was referred to an oncology unit for further management. After three months the patient had extensive metastasis and succumbed to the disease. This case report highlights the clinical presentation, rapid and aggressive course of cardiac angiosarcomas. PMID- 18381878 TI - Hancock valve deterioration in tricuspid position for Ebstein's anomaly. AB - A 65-year-old woman with a Hancock valve implanted 25 years earlier for Ebstein's anomaly underwent a successful second tricuspid valve replacement with a Mosaic valve because of significant tricuspid regurgitation. At surgery, it was found that the Hancock valve had a cylinder-shaped hole and had lost its entire structure. Tricuspid valve dysfunction may be tolerated for a long time before surgery is contemplated. PMID- 18381879 TI - Left heart pump-assisted beating heart coronary surgery in high-risk patients. AB - A simple technique of left ventricular assistance, offering the advantages of both cardiopulmonary bypass and off-pump revascularization, was adopted for high risk patients. It was used in 56 patients with critical left main stenosis and occluded right coronary artery, severely reduced ejection fraction and/or unstable angina. All patients underwent complete and successful myocardial revascularization (3.4 grafts per patient). Weaning from the pump was uncomplicated, and none required conversion to full cardiopulmonary bypass. PMID- 18381880 TI - Easy technique for placing anchoring sutures for aortic root reimplantation. AB - In the aortic valve-sparing reimplantation technique, insertion of the anchoring sutures beneath the valve is a crucial but difficult step because the spared aortic cusps obscure the field of view. We present a novel and easy method of placing these anchoring stitches with good exposure of the subvalvular tissue. PMID- 18381881 TI - Current status of off-pump coronary artery bypass surgery. AB - The expanding indications for angioplasty coupled with the successful short and mid-term results of randomized controlled trials of drug-eluting stents have already had an unquestionable impact on the practice of coronary revascularization operations. However, coronary artery bypass grafting remains a major mode of therapy for coronary artery disease. It is likely that surgery will continue to be preferred for more complex subsets and that surgeons will have to continue to maintain good results in patients with more complex problems. Concerns regarding morbidity associated with conventional surgical myocardial revascularization on cardiopulmonary bypass have led to a resurgence of interest in off-pump bypass surgery during the last decade, with the expectation that it would be safer if cardiopulmonary bypass could be avoided. This review summarizes the impact of off-pump bypass surgery in reducing the morbidity and mortality associated with conventional coronary artery bypass on cardiopulmonary bypass by evaluating the current best-available evidence from randomized controlled trials and meta-analyses comparing off-pump surgery with conventional bypass grafting. PMID- 18381882 TI - Postinfarction giant pseudoaneurysm in the inferior wall of the left ventricle. PMID- 18381883 TI - A large horse shoe-shaped leiomyoma of the thoracic esophagus. PMID- 18381884 TI - Right ventricular perforation induced by trans venous pacing lead. PMID- 18381885 TI - Bilateral common carotid artery aneurysm in Takayasu's arteritis. PMID- 18381886 TI - Evaluating the SF-36 Health Survey (Version 2) in Older Vietnamese Americans. AB - OBJECTIVE: The SF-36((R)) Health Survey (Version 2; SF-36) was evaluated among older Vietnamese Americans to determine whether underlying dimensions of physical and mental health were similar to those of other groups in the United States. METHOD: Field testing of participants from senior centers. RESULTS: The study provided support for the reliability and validity of the SF-36. Structural equation modeling provided confirmation of physical and mental health factors. However, the factor loadings for the SF-36 scales were more consistent with previous results from Asian countries than the typical pattern observed in the United States. DISCUSSION: As the older populations in the United States become more diverse, it is important to have standardized health-related quality of life measures. However, the conceptualization of physical and mental health and associations among different scales may be different for Asian immigrants than for other groups. Thus, the interpretation of the SF-36 scores needs to account for cultural differences. PMID- 18381887 TI - The green yeast uses its plant-like clock to regulate its animal-like tail. PMID- 18381889 TI - A topoisomerase II-dependent mechanism for resetting replicons at the S-M-phase transition. AB - Topoisomerase II (topo II) is required for chromosome segregation and for reprogramming replicons. Here, we show that topo II couples DNA replication termination with the clearing of replication complexes for resetting replicons at mitosis. Topo II inhibition impairs completion of DNA replication, accounting for replication protein A (RPA) stabilization onto ssDNA. Topo II inhibition does not affect the caffeine-sensitive ORC1 degradation found upon origin firing, but it impairs the cdk-dependent degradation/chromatin dissociation of an ORC1/2 reservoir at mitosis. Our results show that ORC1 degradation is rescued by Pin1 depletion and that this topo II-dependent clearing of ORC1/2 from chromatin involves the APC. PMID- 18381890 TI - WD40 protein FBW5 promotes ubiquitination of tumor suppressor TSC2 by DDB1-CUL4 ROC1 ligase. AB - Tuberous sclerosis (TSC) is an autosomal dominant disease characterized by hamartoma formation in various organs and is caused by mutations targeting either the TSC1 or TSC2 genes. TSC1 and TSC2 proteins form a functionally interdependent dimeric complex. Phosphorylation of either TSC subunit by different kinases regulates the function of TSC and represents a major mechanism to integrate various signals into a centralized cell growth pathway. The majority of disease associated mutations targeting either TSC1 or TSC2 results in a substantial decrease in protein level, suggesting that protein turnover also plays a critical role in TSC regulation. Here we report that TSC2 protein binds to FBW5, a DDB1 binding WD40 (DWD) protein, and is recruited by FBW5 to the DDB1-CUL4-ROC1 E3 ubiquitin ligase. Overexpression of FBW5 or CUL4A promotes TSC2 protein degradation, and this is abrogated by the coexpression of TSC1. Conversely, depletion of FBW5, DDB1, or CUL4A/B stabilizes TSC2. Ddb1 or Cul4 mutations in Drosophila result in Gigas/TSC2 protein accumulation and cause growth defects that can be partially rescued by Gigas/Tsc2 reduction. These results indicate that FBW5-DDB1-CUL4-ROC1 is an E3 ubiquitin ligase regulating TSC2 protein stability and TSC complex turnover. PMID- 18381888 TI - Nuclear lamins: major factors in the structural organization and function of the nucleus and chromatin. AB - Over the past few years it has become evident that the intermediate filament proteins, the types A and B nuclear lamins, not only provide a structural framework for the nucleus, but are also essential for many aspects of normal nuclear function. Insights into lamin-related functions have been derived from studies of the remarkably large number of disease-causing mutations in the human lamin A gene. This review provides an up-to-date overview of the functions of nuclear lamins, emphasizing their roles in epigenetics, chromatin organization, DNA replication, transcription, and DNA repair. In addition, we discuss recent evidence supporting the importance of lamins in viral infections. PMID- 18381891 TI - Structure-system correlation identifies a gene regulatory Mediator submodule. AB - A combination of crystallography, biochemistry, and gene expression analysis identifies the coactivator subcomplex Med8C/18/20 as a functionally distinct submodule of the Mediator head module. Med8C forms a conserved alpha-helix that tethers Med18/20 to the Mediator. Deletion of Med8C in vivo results in dissociation of Med18/20 from Mediator and in loss of transcription activity of extracts. Deletion of med8C, med18, or med20 causes similar changes in the yeast transcriptome, establishing Med8C/18/20 as a predominantly positive, gene specific submodule required for low transcription levels of nonactivated genes, including conjugation genes. The presented structure-based system perturbation is superior to gene deletion analysis of gene regulation. PMID- 18381892 TI - Xenopus Paraxial Protocadherin regulates morphogenesis by antagonizing Sprouty. AB - Xenopus Paraxial Protocadherin (xPAPC) has signaling functions that are essential for convergent extension (CE) movements and tissue separation during gastrulation. PAPC modulates components of the planar cell polarity (PCP) pathway, but it is not clear how PAPC is connected to beta-catenin-independent Wnt-signaling. By yeast two-hybrid screen, we found that the intracellular domain of PAPC interacts with Sprouty (Spry), an inhibitor of CE movements. Upon binding to PAPC, Spry function is inhibited and PCP signaling is enhanced. Our data indicate that PAPC promotes gastrulation movements by sequestration of Spry and reveal a novel mechanism by which protocadherins modulate beta-catenin independent Wnt-signaling. PMID- 18381893 TI - The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. AB - Cancer progression has similarities with the process of epithelial-to-mesenchymal transition (EMT) found during embryonic development, during which cells down regulate E-cadherin and up-regulate Vimentin expression. By evaluating the expression of 207 microRNAs (miRNAs) in the 60 cell lines of the drug screening panel maintained by the Nation Cancer Institute, we identified the miR-200 miRNA family as an extraordinary marker for cells that express E-cadherin but lack expression of Vimentin. These findings were extended to primary ovarian cancer specimens. miR-200 was found to directly target the mRNA of the E-cadherin transcriptional repressors ZEB1 (TCF8/deltaEF1) and ZEB2 (SMAD-interacting protein 1 [SIP1]/ZFXH1B). Ectopic expression of miR-200 caused up-regulation of E cadherin in cancer cell lines and reduced their motility. Conversely, inhibition of miR-200 reduced E-cadherin expression, increased expression of Vimentin, and induced EMT. Our data identify miR-200 as a powerful marker and determining factor of the epithelial phenotype of cancer cells. PMID- 18381894 TI - DNA methylation of retrotransposon genes is regulated by Piwi family members MILI and MIWI2 in murine fetal testes. AB - Silencing of transposable elements occurs during fetal gametogenesis in males via de novo DNA methylation of their regulatory regions. The loss of MILI (miwi-like) and MIWI2 (mouse piwi 2), two mouse homologs of Drosophila Piwi, activates retrotransposon gene expression by impairing DNA methylation in the regulatory regions of the retrotransposons. However, as it is unclear whether the defective DNA methylation in the mutants is due to the impairment of de novo DNA methylation, we analyze DNA methylation and Piwi-interacting small RNA (piRNA) expression in wild-type, MILI-null, and MIWI2-null male fetal germ cells. We reveal that defective DNA methylation of the regulatory regions of the Line-1 (long interspersed nuclear elements) and IAP (intracisternal A particle) retrotransposons in the MILI-null and MIWI2-null male germ cells takes place at the level of de novo methylation. Comprehensive analysis shows that the piRNAs of fetal germ cells are distinct from those previously identified in neonatal and adult germ cells. The expression of piRNAs is reduced under MILI- and MIWI2-null conditions in fetal germ cells, although the extent of the reduction differs significantly between the two mutants. Our data strongly suggest that MILI and MIWI2 play essential roles in establishing de novo DNA methylation of retrotransposons in fetal male germ cells. PMID- 18381895 TI - The malate-aspartate NADH shuttle components are novel metabolic longevity regulators required for calorie restriction-mediated life span extension in yeast. AB - Recent studies suggest that increased mitochondrial metabolism and the concomitant decrease in NADH levels mediate calorie restriction (CR)-induced life span extension. The mitochondrial inner membrane is impermeable to NAD (nicotinamide adenine dinucleotide, oxidized form) and NADH, and it is unclear how CR relays increased mitochondrial metabolism to multiple cellular pathways that reside in spatially distinct compartments. Here we show that the mitochondrial components of the malate-aspartate NADH shuttle (Mdh1 [malate dehydrogenase] and Aat1 [aspartate amino transferase]) and the glycerol-3 phosphate shuttle (Gut2, glycerol-3-phosphate dehydrogenase) are novel longevity factors in the CR pathway in yeast. Overexpressing Mdh1, Aat1, and Gut2 extend life span and do not synergize with CR. Mdh1 and Aat1 overexpressions require both respiration and the Sir2 family to extend life span. The mdh1Deltaaat1Delta double mutation blocks CR-mediated life span extension and also prevents the characteristic decrease in the NADH levels in the cytosolic/nuclear pool, suggesting that the malate-aspartate shuttle plays a major role in the activation of the downstream targets of CR such as Sir2. Overexpression of the NADH shuttles may also extend life span by increasing the metabolic fitness of the cells. Together, these data suggest that CR may extend life span and ameliorate age associated metabolic diseases by activating components of the NADH shuttles. PMID- 18381896 TI - Control of cell fate by the formation of an architecturally complex bacterial community. AB - Bacteria form architecturally complex communities known as biofilms in which cells are held together by an extracellular matrix. Biofilms harbor multiple cell types, and it has been proposed that within biofilms individual cells follow different developmental pathways, resulting in heterogeneous populations. Here we demonstrate cellular differentiation within biofilms of the spore-forming bacterium Bacillus subtilis, and present evidence that formation of the biofilm governs differentiation. We show that motile, matrix-producing, and sporulating cells localize to distinct regions within the biofilm, and that the localization and percentage of each cell type is dynamic throughout development of the community. Importantly, mutants that do not produce extracellular matrix form unstructured biofilms that are deficient in sporulation. We propose that sporulation is a culminating feature of biofilm formation, and that spore formation is coupled to the formation of an architecturally complex community of cells. PMID- 18381897 TI - Extensive variation between inbred mouse strains due to endogenous L1 retrotransposition. AB - Numerous inbred mouse strains comprise models for human diseases and diversity, but the molecular differences between them are mostly unknown. Several mammalian genomes have been assembled, providing a framework for identifying structural variations. To identify variants between inbred mouse strains at a single nucleotide resolution, we aligned 26 million individual sequence traces from four laboratory mouse strains to the C57BL/6J reference genome. We discovered and analyzed over 10,000 intermediate-length genomic variants (from 100 nucleotides to 10 kilobases), distinguishing these strains from the C57BL/6J reference. Approximately 85% of such variants are due to recent mobilization of endogenous retrotransposons, predominantly L1 elements, greatly exceeding that reported in humans. Many genes' structures and expression are altered directly by polymorphic L1 retrotransposons, including Drosha (also called Rnasen), Parp8, Scn1a, Arhgap15, and others, including novel genes. L1 polymorphisms are distributed nonrandomly across the genome, as they are excluded significantly from the X chromosome and from genes associated with the cell cycle, but are enriched in receptor genes. Thus, recent endogenous L1 retrotransposition has diversified genomic structures and transcripts extensively, distinguishing mouse lineages and driving a major portion of natural genetic variation. PMID- 18381898 TI - A new way to explore the world of extracellular protein interactions. AB - Eukaryotic genomes encode large numbers of proteins that are either secreted or have exposed extracellular domains. It is highly likely that these proteins facilitate many important biological processes: however, as yet, most remain uncharacterized. Progress in this area of research has been impaired by the lack of a robust screening system that can be used to investigate interactions between large numbers of different extracellular proteins. In this issue, Bushell et al. introduce AVEXIS (avidity-based extracellular interaction screen), a high throughput screening procedure, which can be used to identify even weak extracellular protein interactions with extremely high confidence. This assay represents an important development in the field of network biology. By combining data from the AVEXIS system with data produced by classical or variant yeast two hybrid methods, it will be possible to assemble binary protein interaction networks that connect extracellular and intracellular processes. This information will dramatically increase our ability to understand a wide range of physiological processes and facilitate the development of better therapeutic strategies. PMID- 18381899 TI - Protein networks in disease. AB - During a decade of proof-of-principle analysis in model organisms, protein networks have been used to further the study of molecular evolution, to gain insight into the robustness of cells to perturbation, and for assignment of new protein functions. Following these analyses, and with the recent rise of protein interaction measurements in mammals, protein networks are increasingly serving as tools to unravel the molecular basis of disease. We review promising applications of protein networks to disease in four major areas: identifying new disease genes; the study of their network properties; identifying disease-related subnetworks; and network-based disease classification. Applications in infectious disease, personalized medicine, and pharmacology are also forthcoming as the available protein network information improves in quality and coverage. PMID- 18381900 TI - Prevalence and correlates of shoplifting in the United States: results from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC). AB - OBJECTIVE: This study presented nationally representative data on the lifetime prevalence, correlates, and comorbidity of shoplifting among adults in the United States. METHOD: Data were derived from a large national sample of the United States population. Face-to-face surveys of more than 43,000 adults ages 18 years and older residing in households were conducted during the 2001-2002 period. Diagnoses of mood, anxiety, and drug disorders as well as personality disorders were based on the Alcohol Use Disorder and Associated Disabilities Interview Schedule-DSM-IV Version. RESULTS: The prevalence of lifetime shoplifting in the U.S. population was 11.3%. Associations between shoplifting and all antisocial behaviors were positive and significant. Besides stealing, the behaviors more strongly associated with shoplifting were making money illegally and scamming someone for money. Strong associations between shoplifting and all 12-month and lifetime comorbid psychiatric disorders were also found. The strongest associations with shoplifting were with disorders often associated with deficits in impulse control, such as antisocial personality disorder, substance use disorders, pathological gambling, and bipolar disorder. High rates of mental health service use were also identified in this population. CONCLUSIONS: Shoplifting is a relatively common behavior. A history of shoplifting is associated with substantial rates of comorbid disorders, psychosocial impairment, and mental health service use. Future research should identify the biological and environmental underpinnings of shoplifting and develop effective screening tools and interventions for individuals with shoplifting problems. PMID- 18381901 TI - Brain and plasma pharmacokinetics of aripiprazole in patients with schizophrenia: an [18F]fallypride PET study. AB - OBJECTIVE: Aripiprazole at clinically effective doses occupies some 90% of striatal dopamine 2 and 3 (D(2)/D(3)) receptors. In order to further characterize its extrastriatal and time-dependent binding characteristics, the authors conducted positron emission tomography (PET) studies with the D(2)/D(3) antagonist [(18)F]fallypride at varying time points after the last aripiprazole administration in patients with schizophrenia. METHOD: Sixteen inpatients with a DSM-IV diagnosis of schizophrenia or schizoaffective disorder receiving treatment with aripiprazole underwent an [(18)F]fallypride PET scan. Receptor occupancy was calculated as the percentage reduction in binding potential relative to unblocked values measured in eight age-matched, medication-free patients with schizophrenia. In addition, aripiprazole serum concentrations were determined as part of a routine therapeutic drug monitoring program in a large group of patients (N=128) treated with aripiprazole. RESULTS: Mean dopamine D(2)/D(3) receptor occupancy was high in all brain regions investigated, with no binding difference across brain regions. Nonlinear regression analysis revealed maximum attainable receptor occupancy (E(max)) values close to saturation. The values for serum concentration predicted to provide 50% of E(max) (EC(50)) were in the range of 5-10 ng/ml in all brain regions. The D(2)/D(3) receptors were completely saturated when serum aripiprazole concentration exceeded 100-150 ng/ml. The mean concentration in the large clinical patient sample was 228 ng/ml (SD=142). CONCLUSIONS: Because of its high affinity for D(2)/D(3) receptors and its long elimination half-life, aripiprazole at clinical doses occupies a high fraction of its target receptor everywhere in the brain. Its dissociation from those receptors is very slow, such that the authors calculate from the results that in patients with serum aripiprazole concentrations in the range typical for clinical practice, D(2)/D(3) receptors must remain nearly saturated for as long as 1 week after the last dose. PMID- 18381902 TI - The anatomy of first-episode and chronic schizophrenia: an anatomical likelihood estimation meta-analysis. AB - OBJECTIVE: The authors sought to map gray matter changes in first-episode schizophrenia and to compare these with the changes in chronic schizophrenia. They postulated that the data would show a progression of changes from hippocampal deficits in first-episode schizophrenia to include volume reductions in the amygdala and cortical gray matter in chronic schizophrenia. METHOD: A systematic search was conducted for voxel-based structural MRI studies of patients with first-episode schizophrenia and chronic schizophrenia in relation to comparison groups. Meta-analyses of the coordinates of gray matter differences were carried out using anatomical likelihood estimation. Maps of gray matter changes were constructed, and subtraction meta-analysis was used to compare them. RESULTS: A total of 27 articles were identified for inclusion in the meta analyses. A marked correspondence was observed in regions affected by both first episode schizophrenia and chronic schizophrenia, including gray matter decreases in the thalamus, the left uncus/amygdala region, the insula bilaterally, and the anterior cingulate. In the comparison of first-episode schizophrenia and chronic schizophrenia, decreases in gray matter volume were detected in first-episode schizophrenia but not in chronic schizophrenia in the caudate head bilaterally; decreases were more widespread in cortical regions in chronic schizophrenia. CONCLUSIONS: Anatomical changes in first-episode schizophrenia broadly coincide with a basal ganglia-thalamocortical circuit. These changes include bilateral reductions in caudate head gray matter, which are absent in chronic schizophrenia. Comparing first-episode schizophrenia and chronic schizophrenia, the authors did not find evidence for the temporolimbic progression of pathology from hippocampus to amygdala, but there was evidence for progression of cortical changes. PMID- 18381903 TI - Efficacy of adjunctive aripiprazole to either valproate or lithium in bipolar mania patients partially nonresponsive to valproate/lithium monotherapy: a placebo-controlled study. AB - OBJECTIVE: The authors evaluated the efficacy and safety of adjunctive aripiprazole in bipolar I patients with mania partially nonresponsive to lithium/valproate monotherapy. METHOD: This multicenter, randomized, placebo controlled study included outpatients experiencing a manic or mixed episode (with or without psychotic features). Patients with partial nonresponse to lithium/valproate monotherapy (defined as a Young Mania Rating Scale total score >/=16 at the end of phases 1 and 2, with a decrease of 35% of the standard uptake value). The goal of the study was the definition of biologically different groups of patients prior to or early during induction therapy, with special emphasis on FDG non-avid tumors. EXPERIMENTAL DESIGN: Based on our data, which was published in 2003, at least 36 patients with metabolic response or FDG non-avid tumors had to be recruited for an analysis of the group of FDG non-avid tumors with sufficient statistical power. Seventy-one patients (32 metabolic nonresponders, 17 metabolic responders, and 22 patients with FDG non-avid tumors) underwent FDG-PET at baseline. In FDG-avid tumors, FDG-PET was repeated 14 days after the initiation of chemotherapy. RESULTS: Metabolic responders (17 of 49) showed a high histopathologic response rate (69%) and a favorable prognosis (median survival not reached), whereas metabolic nonresponders (32 of 49) had a poor prognosis (median survival, 24.1 months) and showed a histopathologic response in 17%. The histopathologic response rate (24%) for FDG-PET non-avid patients showed no significant difference compared with FDG-avid nonresponders (P=0.72). Survival of FDG non-avid patients was 36.7 months (not significantly different from FDG-avid nonresponders, 24.1 months, P=0.46). CONCLUSION: In locally advanced gastric cancer, three different metabolic groups exist. Response and survival was predicted by PET in FDG-avid tumors. Metabolic response assessment was not possible in FDG non-avid tumors; however, due to unfavorable outcome, therapy modification might also be considered in FDG non-avid tumors. PMID- 18381940 TI - New pansomatostatin ligands and their chelated versions: affinity profile, agonist activity, internalization, and tumor targeting. AB - PURPOSE: Somatostatin receptor (sst) targeting is an established method to image and treat sst-positive tumors. Particularly, neuroendocrine tumors express the receptor subtype 2 in high density, but sst1, sst3, sst4, and sst5 are also expressed to some extent in different human tumors. Currently used targeting peptides mainly have sst2 affinity. We aimed at developing (radio)peptides that bind with high affinity to all receptor subtypes. EXPERIMENTAL DESIGN: Carbocyclic octapeptides were coupled with macrocyclic chelators for radiometal labeling. Affinity, internalization, and agonist potencies were determined on sst1- to sst5-expressing cell lines. Biodistribution was determined on nude mice bearing HEK-sst2 or AR4-2J and HEK-sst3 tumors. RESULTS: High affinity to all receptor subtypes was found. Y(III)-KE88 showed agonistic properties at all five sst receptor subtypes as it inhibits forskolin-stimulated cyclic AMP production. Surprisingly, very low or even absent sst2 receptor internalization was found compared with currently clinically established octapeptides, whereas the sst3 internalization was very efficient. Biodistribution studies of [(111)In]KE88 and [(67)Ga]KE88/[(68)Ga]KE88 reflected the in vitro data. In nude mice with s.c. implanted sst2 (HEK-sst2, AR4-2J)-expressing and sst3 (HEK-sst3)-expressing tumors, high and persistent uptake was found in sst3-expressing tumors, whereas the uptake in the sst2-expressing tumors was lower and showed fast washout. The kidney uptake was high but blockable by coinjection of lysine. CONCLUSION: This peptide family shows pansomatostatin potency. As radiopeptides, they are the first to show a full pansomatostatin profile. Despite some drawback, they should be useful for imaging sst2-expressing tumors with short-lived radiometals, such as (68)Ga, at early time points and for sst3-expressing tumors at later time points with longer-lived radiometals, such as (64)Cu or (86)Y. PMID- 18381941 TI - Human leukocyte antigen class I, MHC class I chain-related molecule A, and CD8+/regulatory T-cell ratio: which variable determines survival of cervical cancer patients? AB - PURPOSE: To investigate the effect of intraepithelial tumor-infiltrating lymphocytes (ieTIL) and their ligands expressed by cervical tumor cells on the outcome of cervical cancer patients. EXPERIMENTAL DESIGN: The prognostic value of ieTILs was investigated in 115 cases of cervical cancer. T-cell subsets, CD57(+) cells, and regulatory T cells (Treg) were enumerated. The associations of these different ieTIL subtypes with human leukocyte antigen (HLA) class I and MHC class I chain-related molecule A (MICA) expression were determined in relation to clinical variables and patient survival. RESULTS: Survival analysis showed that a high number of intraepithelial Treg (FoxP3(+)), a low CD8(+)/regulatory T-cell ratio, and a weak HLA-A expression were all associated with worse survival (P=0.034, 0.025, and 0.033, respectively, log-rank test). Further stratification of patient groups based on HLA-A-MICA expression and HLA-A-MICA-CD8(+)/Treg ratio revealed an even poorer survival (P=0.005). In a multivariate Cox analysis, low CD8(+)/Treg ratio (P=0.047), weak HLA-A-MICA expression (P=0.003), and weak HLA-A MICA expression combined with low CD8(+)/Treg ratio (P=0.002) were all found to be independent unfavorable prognostic predictors in cervical carcinoma (hazard ratios, 2.7, 4.0, and 4.9, respectively). CONCLUSION: Weak HLA-A-MICA expression combined with low CD8(+)/Treg ratio reveals a patient group with the poorest survival in cervical cancer. As a single variable, low CD8(+)/Treg ratio was a significant independent unfavorable prognostic factor. PMID- 18381942 TI - 18F-fluoro-2-deoxy-glucose uptake predicts clinical outcome in patients with gefitinib-treated non-small cell lung cancer. AB - PURPOSE: To evaluate response and survival according to (18)F-fluoro-2-deoxy glucose uptake at presentation in patients with gefitinib-treated non-small cell lung cancer. EXPERIMENTAL DESIGN: We retrospectively analyzed 84 positron emission tomography/computed tomography findings. Patient characteristics, response rates, and survivals were evaluated according to the maximum standardized uptake value (SUV) of primary tumor. The cutoff value of SUVs was obtained from receiver operating characteristic analysis. RESULTS: The response rate (RR) was higher for never-smokers (41%) than ever-smokers (9%; P=0.001). Patients with adenocarcinoma showed higher RR than those with other tumor histopathology (35% versus 9%; P=0.009). The SUV was significantly lower in patients who were never-smokers (P=0.005), patients with adenocarcinoma (P<0.001), and female patients (P=0.017). Patients with a low SUV showed higher RR compared with those with a high SUV (53% versus 18%; P=0.003). Prolonged progression-free survival was observed in patients with low SUVs compared with those with high SUVs (median, 33.1 weeks versus 8.6 weeks; P=0.003). While controlling for performance status, smoking history, and pathology, the high SUV conferred unfavorable outcome (hazard ratio, 2.3; P=0.012). In terms of overall survival, a low SUV was associated with favorable outcome in univariate analysis (P=0.011). Patients with a low SUV showed prolonged survival in multivariate analysis (P=0.043). CONCLUSIONS: These results suggest that low SUVs at presentation can predict favorable response and survival in gefitinib-treated non small cell lung cancer patients. PMID- 18381944 TI - Glioma proliferation as assessed by 3'-fluoro-3'-deoxy-L-thymidine positron emission tomography in patients with newly diagnosed high-grade glioma. AB - PURPOSE: The aim of this study was to investigate the relationship between the in vivo derived kinetic parameters of 3'-deoxy-3'-(18)F-fluorothymidine ((18)F-FLT) and the proliferation rate measured in vitro by Ki-67 staining in patients with newly diagnosed high-grade gliomas. EXPERIMENTAL DESIGN: Thirteen patients with newly diagnosed high-grade gliomas were investigated with (18)F-FLT and methyl (11)C- l-methionine ((11)C-MET) positron emission tomography (PET) and T1-, Gd-T1 , and T2-weighted magnetic resonance imaging on consecutive days. Tracer kinetic parameters of (18)F-FLT as well as the standardized uptake value and the tumor-to background (T/B) ratio of (18)F-FLT and (11)C-MET were determined. Data of kinetic modeling, standardized uptake value, and T/B values derived from (18)F FLT-PET were compared with T/B values derived from (11)C-MET-PET and to the in vitro proliferation marker Ki-67. RESULTS: A significant correlation was observed between the metabolic rate constant Ki and the proliferation index as measured by Ki-67 immunostaining [Ki, r=0.79 (P=0.004)]. Also, the phosphorylation rate constant k3 correlated with Ki-67 [k3, r=0.76 (P=0.006)], whereas the rate constant for transport through the blood brain barrier K1 showed a weaker correlation with Ki-67 [K1, r=0.62 (P=0.044)]. No significant correlation between (11)C-MET and (18)F-FLT uptake ratios and Ki-67 was observed. CONCLUSIONS: This study shows that kinetic analysis of (18)F-FLT tracer uptake is essential for the in vivo assessment of tumor proliferation in high-grade gliomas, whereas uptake ratios of (11)C-MET and (18)F-FLT failed to correlate with the in vitro determined proliferation marker. Thus, kinetic analysis of (18)F-FLT might provide an accurate method for the assessment of early response to glioma treatment in the future. PMID- 18381943 TI - Single-nucleotide polymorphisms of DNA damage response genes are associated with overall survival in patients with pancreatic cancer. AB - PURPOSE: The goals of this study were to determine if single-nucleotide polymorphisms in DNA damage repair genes and cell cycle regulating genes affect clinical response to combined gemcitabine radiation therapy and the overall survival (OS) of patients with pancreatic cancer. EXPERIMENTAL DESIGN: We evaluated six single-nucleotide polymorphisms of the ATM, ATM and Rad3-related (ATR), CHEK1, and CHEK2 genes in 119 patients with potentially resectable pancreatic cancer who were enrolled in clinical trials at The University of Texas M. D. Anderson Cancer Center from February 1999 to January 2006, with follow-up until February 2007. Patients received neoadjuvant concurrent gemcitabine and radiation therapy with or without gemcitabine-cisplatin induction therapy. Genotypes were determined and tested for associations with OS by Kaplan-Meier estimation, the log-rank test, and Cox regression analysis. P values of 6 months. TKI258 pharmacokinetics were linear over the dose range of 25 to 175 mg. Five of 14 evaluable patients had modulation of phosphorylated ERK levels. CONCLUSIONS: The MTD was defined as 125 mg/d. Evidence of antitumor activity in melanoma and gastrointestinal stromal tumors warrants further investigation, and other phase I studies are ongoing. Further pharmacodynamic evaluation is required in these studies to evaluate the biological effects of TKI258. PMID- 18381948 TI - Anemia is a significant prognostic factor in local relapse-free survival of premenopausal primary breast cancer patients receiving adjuvant cyclophosphamide/methotrexate/5-fluorouracil chemotherapy. AB - PURPOSE: To determine the effects of anemia on local relapse-free, relapse-free, and overall survival (LRFS, RFS, and OS, respectively) in premenopausal, primary breast cancer patients receiving adjuvant polychemotherapy, and to determine which conventional prognostic factors affected these outcomes. EXPERIMENTAL DESIGN: Four hundred twenty-four premenopausal patients with early-stage primary breast cancer and hormone receptor-expressing tumors were treated with i.v. cyclophosphamide/methotrexate/5-fluorouracil (CMF) polychemotherapy as part of an adjuvant phase III trial (Austrian Breast and Colorectal Cancer Study Group Trial 5). The influence of anemia (hemoglobin <12 g/dL) on LRFS, RFS, and OS was evaluated in a retrospective analysis. RESULTS: Of 424 patients, 77 (18.2%) developed anemia on CMF chemotherapy. After a median follow-up time of 5 years, 8.9% of nonanemic patients had local relapse compared with 19.6% of anemic patients (P=0.0006). Although mastectomy was associated with anemia (26% versus 13.7% in breast conserving surgery; P=0.002), multivariate analysis did not show mastectomy per se to be a significant risk factor for LRFS. Age, lymph node status, and hemoglobin had an independent significant influence on LRFS (P<0.005). Anemic patients had a relative risk of 2.96 (95% confidence interval, 1.41-6.23) for developing local relapse in comparison with nonanemic patients. CONCLUSION: Premenopausal breast cancer patients who developed anemia during the CMF regimen had significantly worse LRFS. In Austrian Breast and Colorectal Cancer Study Group Trial 5, anemia may have contributed to an almost doubled incidence of local recurrence in the chemotherapy arm. Molecular targets associated with tumor hypoxia and distinct from erythropoiesis should receive further attention in experimental and clinical settings. PMID- 18381949 TI - The potential predictive value of cyclooxygenase-2 expression and increased risk of gastrointestinal hemorrhage in advanced non-small cell lung cancer patients treated with erlotinib and celecoxib. AB - PURPOSE: Celecoxib, a cyclooxygenase-2 (COX-2) inhibitor, potentiates antitumor effects of erlotinib in preclinical studies, and COX-2 is frequently expressed in non-small cell lung cancer (NSCLC). With these observations, we designed a phase II trial to evaluate the efficacy and safety of erlotinib plus celecoxib in advanced NSCLC. EXPERIMENTAL DESIGN: Previously treated stage IIIB/IV NSCLC patients were given celecoxib at 400 mg orally twice daily and erlotinib at 150 mg orally daily until disease progression. Planned accrual was 40 patients. Tissue was collected for epidermal growth factor receptor (EGFR) analysis and COX 2 immunohistochemistry. RESULTS: Twenty-six patients were enrolled (17 men, 9 women; median age, 66 years). Eighteen and 21 patients had tissue available for EGFR analysis and COX-2 immunohistochemistry, respectively. The median progression-free survival (PFS) and overall survival were 2.0 and 9.2 months, respectively. Eleven of 21 patients tested had increased tumor COX-2 expression, which was strongly associated with prolonged PFS (P=0.048). Four patients on anticoagulation or with a history of peptic ulcer disease had grade 3/grade 4 upper gastrointestinal bleeding (GIB), prompting early study closure. Three patients with GIB had endoscopy that found peptic ulcers. CONCLUSIONS: The combination of erlotinib and celecoxib does not seem superior to erlotinib alone in unselected patients. However, longer PFS with high-tumor COX-2 expression suggests that trials of EGFR and COX-2 inhibitors may be warranted in this patient subset. GIB observed in our trial supports excluding patients with a history of peptic ulcer disease or those requiring therapeutic anticoagulation from future EGFR and COX-2 inhibitor studies. PMID- 18381950 TI - Pilot randomized phase II study of celecoxib in oral premalignant lesions. AB - PURPOSE: Cyclooxygenase-2 (COX-2)-specific inhibition suppresses carcinogenesis in preclinical models and is a promising strategy for preventing oral cancer. In this pilot randomized phase II study, we evaluated the efficacy and safety of the COX-2 inhibitor celecoxib in patients with oral premalignant lesions (OPL). EXPERIMENTAL DESIGN: Patients were randomly assigned to placebo (n=18), celecoxib 100 mg twice daily (n=17), or celecoxib 200 mg twice daily (n=15) for 12 weeks. Six additional patients received celecoxib (400 mg twice daily) in an unblinded extension of the study. Biopsies were obtained at baseline and week 12. All patients entering the study were required to have at least one histologically confirmed early (atypical hyperplasia, atypical hyperkeratosis, or mild dysplasia) or advanced (moderate to severe dysplasia) OPL. RESULTS: Forty-nine patients (46 of 50 randomized and 3 of 6 open label) were evaluable for efficacy analyses. There were no statistically significant differences between the response rates of the randomly assigned arms: placebo, 33.3% (6 of 18); celecoxib 100 mg twice daily, 41.2% (7 of 17); and celecoxib 200 mg twice daily, 20.0% (3 of 15). Two patients responded on celecoxib 400 mg twice daily. Celecoxib was generally well tolerated. Patients with higher baseline COX-2 mRNA levels had an increased risk of disease progression within 3 months. CONCLUSIONS: Celecoxib at 100 or 200 mg twice daily was ineffective in controlling OPLs in this randomized controlled trial. This result and cardiovascular toxicity results of other (large scale) randomized controlled trials of selective COX-2 inhibitors have discouraged the continued investigation of these agents in oral cancer chemoprevention. Better methods for identifying high-risk patients and more active interventions are needed for future oral cancer chemoprevention trials. PMID- 18381951 TI - Population pharmacokinetic-pharmacodynamic model of the vascular-disrupting agent 5,6-dimethylxanthenone-4-acetic acid in cancer patients. AB - PURPOSE: To develop a population pharmacokinetic-pharmacodynamic (PK-PD) model that defines the dose-concentration-effect relationship of 5,6-dimethylxanthenone 4-acetic acid (DMXAA), using plasma 5-hydroxyindole-3-acetic acid (5-HIAA) as a biomarker for the antivascular effect of DMXAA. EXPERIMENTAL DESIGN: The plasma DMXAA and 5-HIAA concentration data were obtained from 124 patients receiving DMXAA monotherapy as a 20-minute i.v. infusion weekly or every 3 weeks at doses of 6 to 4,900 mg/m(2). The PK and PD data were analyzed by nonlinear mixed effects modeling with NONMEM version 5. RESULTS: DMXAA concentration-time profiles were well described by a three-compartment model with saturable elimination (Michaelis-Menten kinetics). Body surface area (BSA) and sex were significant covariates on the volume of distribution of the central compartment (V(1)) and the maximum elimination rate (V(m)), respectively. Population estimates for V(m), K(m) (concentration at which half V(m) is achieved), and V(1) were 112[1 + 0.474(2-sex)] micromol/L/h, 102 micromol/L, and 8.19(BSA/1.8)(0.857) liters, respectively (sex in V(m) is equal to 1 for males and equal to 2 for females). The effect of DMXAA on plasma 5-HIAA was described by the stimulatory E(max) model, where population estimates for baseline, E(max), and EC(50) were 46.3 micromol/L, 2.62-fold increase of the baseline value, and 631 micromol/L, respectively. CONCLUSIONS: DMXAA plasma disposition is characterized by a saturable elimination process. BSA-guided dosing is important. The present PK-PD model, with 5-HIAA as a biomarker, supports the use of DMXAA doses of 1,000 to 2,000 mg/m(2) in phase II studies, and provides an example of how PK-PD models can be used to aid in selection of drug doses for phase II evaluation. PMID- 18381952 TI - Stress, immunity, and cervical cancer: biobehavioral outcomes of a randomized clinical trial [corrected]. AB - PURPOSE: Cancer diagnosis and treatment imparts chronic stressors affecting quality of life (QOL) and basic physiology. However, the capacity to increase survival by improving QOL is controversial. Patients with cervical cancer, in particular, have severely compromised QOL, providing a population well-suited for the evaluation of novel psychosocial interventions and the exploration of mechanisms by which modulation of the psychoneuroimmune axis might result in improved clinical outcomes. EXPERIMENTAL DESIGN: A randomized clinical trial was conducted in cervical cancer survivors that were enrolled at >or=13 and <22 months after diagnosis (n=50), comparing a unique psychosocial telephone counseling (PTC) intervention to usual care. QOL and biological specimens (saliva and blood) were collected at baseline and 4 months post-enrollment. RESULTS: The PTC intervention yielded significantly improved QOL (P=0.011). Changes in QOL were significantly associated with a shift of immune system T helper type 1 and 2 (Th1/Th2) bias, as measured by IFN-gamma/interleukin-5 ELISpot T lymphocyte precursor frequency; improved QOL being associated with increased Th1 bias (P=0.012). Serum interleukin-10 and the neuroendocrine variables of cortisol and dehydroepiandrosterone revealed trends supporting this shift in immunologic stance and suggested a PTC-mediated decrease of the subject's chronic stress response. CONCLUSIONS: This study documents the utility of a unique PTC intervention and an association between changes in QOL and adaptive immunity (T helper class). These data support the integration of the chronic stress response into biobehavioral models of cancer survivorship and suggests a novel mechanistic hypotheses by which interventions leading to enhanced QOL could result in improved clinical outcome including survival. PMID- 18381953 TI - Lupeol inhibits growth of highly aggressive human metastatic melanoma cells in vitro and in vivo by inducing apoptosis. AB - PURPOSE: Poor prognosis of metastatic melanoma mandates the development of novel strategies for its treatment and prevention. In this study, the effect of lupeol, a diet-based triterpene, was determined on the growth and tumorigenicity of human melanoma cells in vitro and in vivo. EXPERIMENTAL DESIGN: Normal human melanocytes, and human metastatic (451Lu) and nonmetastatic (WM35) cells were treated with lupeol; its effect on growth, proliferation, and apoptosis were evaluated. Further athymic nude mice bearing 451Lu cell-originated tumors were administered with lupeol thrice a week, and its effect on tumor growth and surrogate biomarkers was evaluated. RESULTS: Lupeol significantly decreased the viability of 451Lu and WM35 melanoma cells but had only a marginal effect on normal human melanocyte cells at similar doses. Lupeol treatment of 451Lu cells caused (a) G(1)-S phase cell cycle arrest and apoptosis; (b) down-regulation of Bcl2 and up-regulation of Bax; (c) activation of caspase-3 and induction of poly(ADP)ribose polymerase cleavage; (d) decreased expression of cyclin D1, cyclin D2, and cdk2; and (e) increased expression of p21 protein. Next, lupeol significantly reduced 451Lu tumor growth in athymic nude mice and modulated the expression of proliferation markers, apoptotic markers, and cell cycle regulatory molecules in tumor xenografts. CONCLUSION: Our findings showed the anticancer efficacy of lupeol with mechanistic rationale against metastatic human melanoma cells. We suggest that lupeol, alone or as an adjuvant to current therapies, could be useful for the management of human melanoma. PMID- 18381954 TI - Curcumin sensitizes human colorectal cancer xenografts in nude mice to gamma radiation by targeting nuclear factor-kappaB-regulated gene products. AB - PURPOSE: How colorectal cancer develops resistance to gamma-radiation is not fully understood, but the transcription factor nuclear factor-kappaB (NF-kappaB) and NF-kappaB-regulated gene products have been proposed as mediators. Because curcumin, a component of turmeric (Curcuma longa), has been shown to suppress NF kappaB activation, whether it can sensitize the colorectal cancer to gamma radiation was investigated in colorectal cancer xenografts in nude mice. EXPERIMENTAL DESIGN: We established HCT 116 xenograft in nude mice, randomized into four groups, and treated with vehicle (corn oil), curcumin, gamma-radiation, and curcumin in combination with gamma-radiation. NF-kappaB modulation was ascertained using electrophoretic mobility shift assay and immunohistochemistry. Markers of proliferation, angiogenesis, and invasion were monitored by immunohistochemistry and Western blot analysis. RESULTS: Curcumin significantly enhanced the efficacy of fractionated radiation therapy by prolonging the time to tumor regrowth (P=0.02) and by reducing the Ki-67 proliferation index (P<0. 001). Moreover, curcumin suppressed NF-kappaB activity and the expression of NF-kappaB regulated gene products (cyclin D1, c-myc, Bcl-2, Bcl-xL, cellular inhibitor of apoptosis protein-1, cyclooxygenase-2, matrix metalloproteinase-9, and vascular endothelial growth factor), many of which were induced by radiation therapy and mediate radioresistance. The combination of curcumin and radiation therapy also suppressed angiogenesis, as indicated by a decrease in vascular endothelial growth factor and microvessel density (P=0.002 versus radiation alone). CONCLUSION: Collectively, our results suggest that curcumin potentiates the antitumor effects of radiation therapy in colorectal cancer by suppressing NF kappaB and NF-kappaB-regulated gene products, leading to inhibition of proliferation and angiogenesis. PMID- 18381955 TI - Pharmacology of the novel antiangiogenic peptide ATN-161 (Ac-PHSCN-NH2): observation of a U-shaped dose-response curve in several preclinical models of angiogenesis and tumor growth. AB - PURPOSE: ATN-161 (Ac-PHSCN-NH(2)) is an integrin-binding peptide that is currently in phase II trials in cancer patients. This peptide has been shown to have antitumor activity in a number of different preclinical models. EXPERIMENTAL DESIGN: In this study, we examined the binding, biodistribution, and dose and biomarker response of ATN-161 in several animal models. RESULTS: ATN-161 bound to the beta subunit of a number of different integrins implicated in tumor growth and progression, which depended on its cysteine thiol. The peptide had antiangiogenic activity in the Matrigel plug model, and this activity could be reversed by inhibitors of protein kinase A, an effector of alpha(5)beta(1) dependent angiogenesis. A labeled analogue of ATN-161, ATN-453, localized to neovessels but not to preexisting vasculature in vivo. The half-life of the peptide when localized to a tumor was much longer than in plasma. Dose-response studies in the Matrigel plug model of angiogenesis or a Lewis lung carcinoma model of tumor growth showed a U-shaped dose-response curve with 1 to 10 mg/kg given thrice a week, being the optimal dose range of ATN-161. Two additional pharmacodynamic models of angiogenesis (dynamic contrast-enhanced magnetic resonance imaging and measurement of endothelial cell progenitors) also revealed U-shaped dose-response curves. CONCLUSIONS: The presence of a U-shaped dose response curve presents a significant challenge to identifying a biologically active dose of ATN-161. However, the identification of biomarkers of angiogenesis that also exhibit this same U-shaped response should allow the translation of those biomarkers to the clinic, allowing them to be used to identify the active dose of ATN-161 in phase II studies. PMID- 18381956 TI - DTS-108, a novel peptidic prodrug of SN38: in vivo efficacy and toxicokinetic studies. AB - PURPOSE: Irinotecan is a prodrug converted to the active cytotoxic molecule SN38 predominantly by the action of liver carboxylesterases. The efficacy of irinotecan is limited by this hepatic activation that results in a low conversion rate, high interpatient variability, and dose-limiting gastrointestinal toxicity. The purpose of this study was to evaluate a novel peptidic prodrug of SN38 (DTS 108) developed to bypass this hepatic activation and thus reduce the gastrointestinal toxicity and interpatient variability compared with irinotecan. EXPERIMENTAL DESIGN: SN38 was conjugated to a cationic peptide (Vectocell) via an esterase cleavable linker. The preclinical development plan consisted of toxicity and efficacy evaluation in a number of different models and species. RESULTS: The conjugate (DTS-108) is highly soluble, with a human plasma half-life of 400 minutes in vitro. Studies in the dog showed that DTS-108 liberates significantly higher levels of free SN38 than irinotecan without causing gastrointestinal toxicity. In addition, the ratio of the inactive SN38-glucuronide metabolite compared with the active SN38 metabolite is significantly lower following DTS-108 administration, compared with irinotecan, which is consistent with reduced hepatic metabolism. In vivo efficacy studies showed that DTS-108 has improved activity compared with irinotecan. A significant dose-dependent antitumoral efficacy was observed in all models tested and DTS-108 showed synergistic effects in combination with other clinically relevant therapeutic agents. CONCLUSIONS: DTS-108 is able to deliver significantly higher levels of SN38 than irinotecan, without the associated toxicity of irinotecan, resulting in an increased therapeutic window for DTS-108 in preclinical models. These encouraging data merit further preclinical and clinical investigation. PMID- 18381957 TI - A severe combined immunodeficient-hu in vivo mouse model of human primary mantle cell lymphoma. AB - PURPOSE: To establish a severe combined immunodeficient (SCID)-hu in vivo mouse model of human primary mantle cell lymphoma (MCL) for the study of the biology and novel therapy of human MCL. EXPERIMENTAL DESIGN: Primary MCL cells were isolated from spleen, lymph node, bone marrow aspirates, or peripheral blood of six different patients and injected respectively into human bone chips, which had been s.c. implanted in SCID-hu. Circulating human beta(2)-microglobulin in mouse serum was used to monitor the engraftment and growth of patient's MCL cells. H&E staining and immunohistochemical staining with anti-human CD20 and cyclin D1 antibodies were used to confirm the tumor growth and migration. RESULTS: Increasing levels of circulating human beta(2)-microglobulin in mouse serum indicated that the patient's MCL cells were engrafted successfully into human bone chip of SCID-hu mice. The engraftment and growth of patient's MCL cells were dependent on human bone marrow microenvironment. Immunohistochemical staining with anti-human CD20 and cyclin D1 antibodies confirmed that patient's MCL cells were able to not only survive and propagate in the bone marrow microenvironment of the human fetal bone chips, but also similar to the human disease, migrate to lymph nodes, spleen, bone marrow, and gastrointestinal tract of host mice. Treatment of MCL-bearing SCID-hu mice with atiprimod, a novel antitumor compound against the protection of bone marrow stromal cells, induced tumor regression. CONCLUSION: This is the first human primary MCL animal model that should be useful for the biological and therapeutic research on MCL. PMID- 18381958 TI - Evaluation of D-methionine as a novel oral radiation protector for prevention of mucositis. AB - PURPOSE: Oral mucositis is a common acute morbidity associated with radiation and/or chemotherapy treatment for cancer. D-Methionine (D-Met), the dextro-isomer of the common amino acid l-methionine, has been documented to protect normal tissues from a diverse array of oxidative insults. EXPERIMENTAL DESIGN: We evaluated if D-Met could selectively prevent radiation-induced oral mucositis using in vitro cell culture models as well as an in vivo model of radiation injury to the oral mucosa in C3H mice. RESULTS: Unlike free-radical scavengers, which protected both normal and transformed tumor cells in vitro from radiation induced cell death, treatment with d-Met in culture protected nontransformed primary human cells from radiation-induced cell death (protective factor between 1.2 and 1.6; P<0.05) whereas it did not confer a similar protection on transformed tumor cells. D-Met treatment also provided significant protection to normal human fibroblasts, but not to tumor cell lines, from radiation-induced loss of clonogenicity (protection factor, 1.6+/-0.15). D-Met treatment did not alter DNA damage (as measured by histone phosphorylation) following irradiation but seemed to selectively mitigate the loss of mitochondrial membrane potential in nontransformed cells, whereas it did not provide a similar protection to tumor cells. Tumor control of implanted xenografts treated with radiation or concurrent cisplatin and radiation was not altered by D-Met treatment. Pharmacokinetics following administration of a liquid suspension of D-Met in rats showed 68% bioavailability relative to i.v. administration. Finally, in a murine model of mucositis, a dose-dependent increase in protection was observed with the protective factor increasing from 1.6 to 2.6 over a range of oral D-Met doses between 200 and 500 mg/kg (P<0.0003). CONCLUSIONS: D-Met protected normal tissues, but not tumor cells, in culture from radiation-induced cell death; it also protected normal cells from radiation-induced mucosal injury in a murine model but did not alter tumor response to therapy. Further studies on the use of D-Met to protect from oral mucositis are warranted. PMID- 18381959 TI - Direct visualization of heterogeneous extravascular distribution of trastuzumab in human epidermal growth factor receptor type 2 overexpressing xenografts. AB - PURPOSE: The high molecular weight and binding affinity of trastuzumab, a monoclonal antibody in use for treatment of breast cancers overexpressing human epidermal growth factor receptor type 2 (HER2), in combination with microenvironmental factors, may limit its distribution and efficacy. We assessed and mapped the distribution of systemically given, unlabeled trastuzumab at micrometer resolution in tumor xenografts using immunohistochemistry. EXPERIMENTAL DESIGN: Mice bearing MDA-435/LCC6(HER2) xenografts were given single doses of 4 or 20 mg/kg unlabeled trastuzumab with tumor harvest at various time points thereafter; bound trastuzumab was imaged directly in tumor cryosections using fluorescently tagged antihuman secondary antibodies. Combinations of additional markers, including HER2, 5-bromo-2-deoxyuridine, CD31, DioC(7)(3), desmin, and collagen IV were also mapped on the same tumor sections. RESULTS: Distribution of trastuzumab in MDA-435/LCC6(HER2) tumors is found to be heterogeneous, with tumor margins saturating more thoroughly in doses and times analyzed. Considerable intervessel heterogeneity is also seen. For example, in unsaturated tissues, there remain perfused vessels without any trastuzumab in addition to vessels with a few layers of positively stained perivascular cells, in addition to vessels with bound drug up to 150 microm away. This heterogeneity is independent of HER2 expression, microvessel density, and perfusion. A slightly greater proportion of vessels were associated with pericytes in sections with greater trastuzumab saturation, but this would not adequately account for observed heterogeneous trastuzumab distribution. CONCLUSIONS: Complete penetration of trastuzumab in tumor tissue was not seen in our study, leaving the possibility that inadequate distribution may represent a mechanism for resistance to trastuzumab. PMID- 18381961 TI - Tumor-targeting nanocomplex delivery of novel tumor suppressor RB94 chemosensitizes bladder carcinoma cells in vitro and in vivo. AB - PURPOSE: RB94, a truncated form of RB110, has enhanced tumor suppressor potency and activity against all tumor types tested to date including bladder carcinoma. However, efficient, systemic delivery of the gene encoding RB94 specifically to tumors, is an obstacle to clinical application as an anticancer therapeutic. We have developed a systemically given, nanosized liposome DNA delivery system that specifically targets primary and metastatic disease. The ability of RB94, delivered via this nanocomplex, to sensitize bladder carcinoma to chemotherapy in vitro and in vivo was assessed. EXPERIMENTAL DESIGN: The nanocomplex is an RB94 plasmid encapsulated by a cationic liposome, the surface of which is decorated with a tumor-targeting moiety, either transferrin (Tf/Lip/RB94) or an antitransferrin receptor single-chain antibody fragment (TfRScFv/Lip/RB94). The ability of the complex to sensitize human bladder carcinoma HTB-9 cells to chemotherapeutics was assessed in vitro by XTT assay. In vivo tumor specificity and efficacy were tested in mice carrying HTB-9 tumors by PCR and tumor growth inhibition, respectively. RESULTS: Transfection with Tf/Lip/RB94 significantly sensitized HTB-9 cells to chemotherapeutic agents in vitro. Tumor specificity of the complex was shown in an orthotopic bladder tumor model by immunohistochemistry and PCR. Moreover, in mice bearing subcutaneous HTB-9 tumors, the combination of systemically given Tf/Lip/RB94 or TfRScFv/Lip/RB94 plus gemcitabine resulted in significant (P<0.0005) tumor growth inhibition/regression and induction of apoptosis. CONCLUSIONS: Use of our tumor targeting nanocomplex to specifically deliver the potent tumor suppressor RB94 efficiently to tumors has potential as a more effective treatment modality for genitourinary and other cancers. PMID- 18381960 TI - Treatment of human colon cancer xenografts with TRA-8 anti-death receptor 5 antibody alone or in combination with CPT-11. AB - PURPOSE: This study was designed to evaluate the in vitro cytotoxicity and in vivo efficacy of TRA-8, a mouse monoclonal antibody that binds to the DR5 death receptor for tumor necrosis factor-related apoptosis-inducing ligand (also called Apo2L), alone and in combination with CPT-11, against human colon cancer cells and xenografts. EXPERIMENTAL DESIGN: DR5 expression was assessed on human colon cancer cell lines using flow cytometry, and cellular cytotoxicity after TRA-8 treatment, alone and in combination with SN-38, was determined by measuring cellular ATP levels. Tumor growth inhibition and regression rates of well established subcutaneous COLO 205, SW948, HCT116, and HT-29 colon cancer xenografts in athymic nude mice treated with TRA-8 or CPT-11 alone and in combination were determined. (99m)Tc-TRA-8 was used to examine tumor localization of TRA-8 in animals bearing each of the four xenografts. In addition, whole-body biodistribution and imaging was carried out in COLO 205-bearing animals using in vivo single-photon emission computed tomography imaging and tissue counting. RESULTS: DR5 expression was highest on HCT116, intermediate on SW948 and COLO 205 cells, and lowest on HT-29 cells. COLO 205 cells were the most sensitive to TRA-8 induced cytotoxicity in vitro, SW948 and HCT116 cell lines were moderately sensitive, and HT-29 cells were resistant. Combination treatment with TRA-8 and SN-38 produced additive to synergistic cytotoxicity against all cell lines compared with either single agent. The levels of apoptosis in all cell lines, including HT-29, were increased by combination treatment with SN-38. In vivo, combination therapy with TRA-8 and CPT-11 was superior to either single-agent regimen for three of the xenografts: COLO 205, SW948, and HCT116. COLO 205 tumors were most responsive to therapy with 73% complete regressions after combination therapy. HT-29 cells derived no antitumor efficacy from TRA-8 therapy. Tumor xenografts established from the four colon cancer cell lines had comparable specific localization of (99m)Tc-TRA-8. CONCLUSIONS: In vitro and in vivo effects of TRA-8 anti-DR5 monoclonal antibody on four different colon cancer cell lines and xenografts were quite variable. The HT-29 cell line had low surface DR5 expression and was resistant to TRA-8 both in vitro and in vivo. Three cell lines (COLO 205, SW948, and HCT116) exhibited moderate to high sensitivity to TRA-8 mediated cytotoxicity which was further enhanced by the addition of SN-38, the active metabolite of CPT-11. In vivo, the combination of TRA-8 and CPT-11 treatment produced the highest antitumor efficacy against xenografts established from the three TRA-8-sensitive tumor cell lines. All four colon cancer xenografts had comparable localization of (99m)Tc-TRA-8. These studies support the strategy of TRA-8/CPT-11 combined treatment in human colon cancer clinical trials. PMID- 18381962 TI - Loss of retinoblastoma tumor suppressor protein makes human breast cancer cells more sensitive to antimetabolite exposure. AB - PURPOSE: The RB tumor-suppressor activity may influence the therapeutic response in human breast cancers. The effect of adjuvant therapy on clinical outcome of breast cancer patients was analyzed, and the sensitivity to 5-fluorouracil (5-FU) and methotrexate was investigated in MCF-7 and HCT-116 human cancer cells, according to their RB status. EXPERIMENTAL DESIGN: RB protein (pRB) expression was prospectively evaluated by immunocytochemistry in 518 consecutive patients and its predictive value was determined according to the adjuvant therapeutic treatments. MCF-7 and HCT-116 human cancer cells silenced for RB1 expression were treated with 5-FU and methotrexate, at the same concentrations and time exposures as determined in the interstitium of breast cancers of patients treated with adjuvant chemotherapy. RESULTS: Multivariate analysis of disease-free survival, including all the established clinical and histopathologic prognostic variables, indicated that the absence of pRB expression was the only predictive factor of good clinical outcome in patients treated with standard systemic chemotherapy (cyclophosphamide, methotrexate, and 5-FU) but not in patients treated with endocrine therapy alone. 5-FU and methotrexate significantly reduced the growth rate of RB1-silenced but not of control MCF-7 and HCT-116 cells. This was likely due to the absence of a DNA damage checkpoint with accumulation of DNA double strand breaks in RB1-silenced but not in control cells. CONCLUSIONS: The absence of pRB expression renders human breast cancer cells more sensitive to 5-FU and methotrexate and predicts a good clinical outcome for patients treated with adjuvant chemotherapy. We suggest that patients with RB-negative breast cancers should be treated with systemic chemotherapy. PMID- 18381963 TI - Combination of vascular endothelial growth factor receptor/platelet-derived growth factor receptor inhibition markedly improves radiation tumor therapy. AB - PURPOSE: Investigations on the combination of radiotherapy with vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) antiangiogenic agents, which has the potential to improve the clinical outcome in cancer patients. EXPERIMENTAL DESIGN: Here, we analyze the combined VEGF (SU5416) and PDGF (SU6668) receptor tyrosine kinase inhibition with irradiation in human endothelium (HUVEC), prostate cancer (PC3), and glioblastoma (U87) in vitro and in vivo. RESULTS: Combined inhibition of VEGF and PDGF signaling resulted in enhanced apoptosis, reduced cell proliferation, and clonogenic survival as well as reduced endothelial cell migration and tube formation compared with single pathway inhibition. These effects were further enhanced by additional irradiation. Likewise, in PC3 and U87 tumors growing s.c. on BALB/c nu/nu mice, dual inhibition of VEGF and PDGF signaling significantly increased tumor growth delay versus each monotherapy. Interestingly, radiation at approximately 20% of the dose necessary to induce local tumor control exerts similar tumor growth inhibitory effects as the antiangiogenic drugs given at their maximum effective dose. Addition of radiotherapy to both mono- as well as dual-antiangiogenic treatment markedly increased tumor growth delay. With respect to tumor angiogenesis, radiation further decreased microvessel density (CD31 count) and tumor cell proliferation (Ki-67 index) in all drug-treated groups. Of note, the slowly growing PC3 tumor responded better to the antiangiogenic drug treatments than the faster-growing U87 tumor. In addition to the beneficial effect of abrogating VEGF survival signaling when combined with radiation, we identified here a novel mechanism for the tumor escape from radiation damage. We found that radiation induced up-regulation of all four isoforms of PDGF (A-D) in endothelial cells supporting adjacent smooth muscle cells resulting in a prosurvival effect of radiation. The addition of SU6668 attenuated this undesirable paracrine radiation effect, which may rationalize the combined application of radiation with PDGF signaling inhibition to increase antitumor effects. CONCLUSION: A relative low radiation dose markedly enhances local antitumor effects of combined VEGF and PDGF signaling inhibition, suggesting a promising combination regimen for local tumor treatment with radiotherapy remaining an essential element. PMID- 18381964 TI - An in vivo model of Met-driven lymphoma as a tool to explore the therapeutic potential of Met inhibitors. AB - PURPOSE: Met, the tyrosine kinase receptor for hepatocyte growth factor, is frequently deregulated in human cancer. Recent evidence indicates that Met amplification may confer resistance to treatments directed toward other receptor tyrosine kinases. Thus, there is a need to develop Met inhibitors into therapeutic tools, to be used alone or in combination with other molecularly targeted drugs. Preclinical validation of Met inhibitors has thus far been done in nude mice bearing cancer cells xenografts. A far superior model would be a transgenic line developing spontaneous Met-driven tumors with high penetrance and short latency. EXPERIMENTAL DESIGN: To this end, we introduced into the mouse genome TPR-MET, the oncogenic form of MET. The Tpr-Met protein ensures deregulation of Met signaling because dimerization motifs in the Tpr moiety cause ligand-independent activation of the Met kinase. RESULTS: Here, we describe a TPR MET transgenic line that develops thymic T-cell lymphoma with full penetrance and very short latency. In the tumors, Tpr-Met and its effectors were phosphorylated. Treatment of tumor-derived T lymphocytes with the selective Met inhibitor PHA 665752 at nanomolar concentrations abolished phosphorylation of Met and downstream effectors and led to caspase-mediated apoptosis. I.v. administration of PHA-665752 to transgenic mice bearing lymphomas in exponential growth phase led to a significant decrease in tumor growth and, in some cases, to tumor regression. CONCLUSIONS: Our transgenic line, which within 2 months reliably develops Tpr-Met-driven T-cell lymphoma, represents a valuable tool to explore the efficacy and therapeutic potential of Met kinase inhibitors as anticancer drugs. PMID- 18381965 TI - Toxigenic Helicobacter pylori infection precedes gastric hypochlorhydria in cancer relatives, and H. pylori virulence evolves in these families. AB - PURPOSE: Helicobacter pylori infection by virulent strains is associated with gastric adenocarcinoma. We aimed to determine whether infection with virulent H. pylori preceded precancerous gastric hypochlorhydria and atrophy in gastric cancer relatives and quantify the extent of virulence factor evolution. EXPERIMENTAL DESIGN: H. pylori strains from 51 Scottish gastric cancer relatives were characterized by genetic fingerprinting and typing the vacuolating cytotoxin gene (vacA), the cytotoxin-associated gene (cagA), and housekeeping genes. We phenotyped strains by coculture with gastric epithelial cells and assessing vacuolation (microscopy), CagA tyrosine phosphorylation (immunoblot), and interleukin-8 secretion (ELISA). RESULTS: Toxigenic (vacA type s1/m1) H. pylori was associated with precancerous gastric hypochlorhydria (P<0.01). Adult family members with this type of H. pylori had the same strain as currently noncohabiting adult family members in 68% cases, implying acquisition during childhood from each other or a common source. We analyzed different isolates of the same strain within families and showed that H. pylori commonly microevolved to change virulence: this occurred in 22% individuals and a striking 44% cases where the strain was shared within families. Microevolution in vacA occurred by extragenomic recombination and in cagA by this or duplication/deletion. Microevolution led to phenotypic changes in virulence. Passage of microevolved strains could be tracked within families. CONCLUSIONS: Toxigenic H. pylori infection precedes and so likely causes gastric hypochlorhydria, suggesting that virulent H. pylori increases cancer risk by causing this condition. Microevolution of virulence genes is common within families of gastric cancer patients and changes H. pylori virulence. PMID- 18381966 TI - Profiling of genetic variations in inflammation pathway genes in relation to bladder cancer predisposition. AB - PURPOSE: Compelling evidence has indicated that inflammation plays an important role in cancer development. We sought to test the hypothesis that common sequence variants in the inflammation pathway modulate bladder cancer risk. EXPERIMENTAL DESIGN: We genotyped 59 potentially functional single nucleotide polymorphisms from 35 candidate inflammation genes in a case-control study including 635 Caucasian bladder cancer patients and 635 matched controls. RESULTS: The most significant finding was in the 3'-untranslated region of PTGS2 (exon10+837T>C, rs5275), which was associated with a significantly reduced risk (odds ratio, 0.68; 95% confidence interval, 0.54-0.87; P=0.002) and remained significant after multiple comparison adjustment. Consistently, the most common PTGS2 haplotype containing the common allele of exon10+837T>C was associated with a significantly increased risk (odds ratio, 1.27; 95% confidence interval, 1.06-1.52; P=0.008). In contrast, the haplotypes containing at least one variant allele of exon10+837T>C were all associated with a decreased risk. In a combined analysis to assess the cumulative effects of inflammation single nucleotide polymorphisms on bladder cancer risk, we found that in the anti-inflammation pathway, but not in the proinflammation pathway, when compared with individuals with a few adverse alleles, individuals with more adverse alleles had a significantly increased risk in a dose-dependent manner (P(trend)=0.012). To further elucidate the functional mechanism of these associations, we redefined the adverse alleles based on literature-reported functional results and found that individuals with a higher number of inflammation-enhancing alleles in the anti-inflammation pathway exhibited a greater bladder cancer risk. CONCLUSIONS: Our results strongly suggest that common variants in inflammation genes affect bladder cancer susceptibility individually and jointly. PMID- 18381968 TI - Re: Wieland BM, et al. Is there a human homologue to the murine proteolysis inducing factor? PMID- 18381969 TI - Setting the stage: possible mechanisms by which acute contraction restores insulin sensitivity in muscle. AB - It has long been known that acute exercise can dramatically improve insulin sensitivity in previously insulin-resistant muscle; however, the precise mechanisms underlying this clinically significant interaction remain unknown. Using hindlimb perfusions in obese Zucker rats, our group found that acute muscle contraction synergistically improved insulin-stimulated glucose transport in skeletal muscle, but contrary to our hypothesis, these findings were not associated with either improved insulin signaling or decreased intramuscular lipid metabolites. A further analysis revealed that the improved insulin sensitivity was associated with a robust increase in mitochondrial energy flux. These findings and reports from other labs suggest that mitochondrial energy flux and mitochondrial oxidative capacity may govern insulin sensitivity and override insulin signaling defects associated with obesity. This review will discuss the effects of acute exercise to enhance insulin sensitivity in previously insulin resistant muscle and present possible novel mechanisms by which alterations in mitochondrial energy metabolism may play a regulatory role. PMID- 18381970 TI - Educating for advocacy: recommendations for professional preparation and development based on a needs and capacity assessment of health education faculty. AB - An electronic survey was used to conduct a needs and capacity assessment of health education faculty to determine the extent to which advocacy instruction is present in undergraduate and graduate curricula in health education and to identify faculty members' needs and capacity to provide professional preparation and development experiences related to advocacy. An analysis of the results reveals that most undergraduate and graduate health education programs include advocacy instruction. Although faculty believe advocacy and instruction related to advocacy are important, many lack advocacy-related professional preparation and development experiences and do not participate in advocacy-related training initiatives and advocacy activities. There is wide variability in faculty confidence in their competence to provide advocacy instruction. Partnerships among professional organizations, health education practitioners, university faculty, individuals engaged in policy advocacy initiatives, and policy makers are needed to enhance the capacity of university faculty to provide professional preparation and development experiences related to advocacy. PMID- 18381971 TI - Assessment of training needs and preferences for geographic information systems (GIS) mapping in state comprehensive cancer-control programs. AB - Geographic information systems (GIS) mapping technologies have potential to advance public health promotion by mapping regional differences in attributes (e.g., disease burden, environmental exposures, access to health care services) to suggest priorities for public health interventions. Training in GIS for comprehensive cancer control (CCC) has been overlooked. State CCC programs' GIS training needs were assessed by interviewing 49 state CCC directors. A majority perceived a need for GIS training, slightly more than half of state CCC programs had access to geocoded data, and the majority of programs did not require continuing education credits of their staff. CCC directors perceived judging maps and realizing their limitations as important skills and identified epidemiologists, CCC staff, public health officials, policy makers, and cancer coalition members as training audiences. They preferred in-class training sessions that last a few hours to a day. Lessons learned are shared to develop training programs with translatable GIS skills for CCC. PMID- 18381972 TI - Comparing the efficacy of permanent classroom teachers to temporary health educators for pregnancy and HIV prevention instruction. AB - Ninth grade health education classrooms were randomly assigned to conditions in which a 14-lesson unit on HIV and pregnancy prevention was provided either by the permanent classroom teacher or by a temporary health educator. Student data were collected prior to the lessons and approximately 3 weeks after the completion of the unit. Results indicate that students who learned the unit from the classroom teacher reported that that the instructor was more likeable and credible and that the classrooms were more motivational. Sexually active adolescents with regular classroom teachers reported greater increases in frequency of class discussion. PMID- 18381973 TI - MMPI-2 profiles in civilian PTSD: an examination of differential responses between victims of crime and industrial accidents. AB - The authors studied MMPI-2 profiles of workers (N = 83) diagnosed with posttraumatic stress disorder (PTSD) and a control group comprising workers with chronic pain (N = 40). Significant differences were seen in profiles between the PTSD groups and the control group, and the authors compared the PTSD profiles according to exposure to two different kinds of traumatic incidents: industrial accidents or criminal events. They found differences in profile elevations based on the type of event: The level of distress and fear is greater in PTSD victims of crime, and the results also suggest that victims of crime are more suspicious and guarded compared with accident victims. Theoretically based reasons for the differences in profile elevations between the two PTSD groups are discussed. PMID- 18381975 TI - Investment analysts and the American Society of Hematology. AB - Investment analysts are a growing presence at the Annual Meeting of the American Society of Hematology (ASH), and financial professionals frequently contact ASH members for information and perspective on drugs, devices, and scientific developments. Recent incidents have raised concerns about consulting relationships between physicians and the investment industry; the appropriate role of medical societies in influencing these relationships is unclear. In this essay, I summarize the current situation, discuss potential risks and benefits from interactions between physicians and investment analysts, and outline issues that all individuals involved in investment industry consulting should consider. I also propose changes in ASH policy that may help safeguard public trust as well as preserve the access of clinicians and scientists to clinically relevant data presented at the Annual Meeting. PMID- 18381974 TI - Persistent inward currents in spinal motoneurons and their influence on human motoneuron firing patterns. AB - Persistent inward currents (PICs) are present in many types of neurons and likely have diverse functions. In spinal motoneurons, PICs are especially strong, primarily located in dendritic regions, and subject to particularly strong neuromodulation by the monoamines serotonin and norepinephrine. Because motoneurons drive muscle fibers, it has been possible to study the functional role of their PICs in motor output and to identify PIC-mediated effects on motoneuron firing patterns in human subjects. The PIC markedly amplifies synaptic input, up to fivefold or more, depending on the level of monoaminergic input. PICs also tend to greatly prolong input time course, allowing brief inputs to initiate long-lasting self-sustained firing (i.e., bistable behavior). PIC deactivation usually requires inhibitory input and PIC amplitude can increase to repeated activation. All of these behaviors markedly increase motoneuron excitability. Thus, in the absence of monoaminergic input, motoneuron excitability is very low. Yet PICs have another effect: once active, they tend to sharply limit efficacy of additional synaptic input. All of these PIC effects have been detected in motoneuron firing patterns in human subjects and, hence, PICs are likely a fundamental component of normal motor output. PMID- 18381977 TI - Egg box conformation of oligogalacturonides: the time-dependent stabilization of the elicitor-active conformation increases its biological activity. AB - Circular dichroism spectrometry was used on oligogalacturonides (OGAs) and showed the existence of a calcium/sodium-induced conformational state that is intermediate between single-isolated chains and calcium-associated multimer chains. This conformation is interpreted as being egg box dimers. Using the 2F4 monoclonal antibody that specifically binds such an egg box dimer conformation of pectin, the stability of OGA dimers was investigated over a period of 24 hours. The extent to which egg box dimers were recognized by the antibody was dependent on the temperature and duration of preincubation of the OGA. This suggests a "maturation" process of the egg-box structure that consists in a progressive increase in the length of the junction sequences between two chains that slide along each other in order to form a maximum number of calcium bridges and dimer ends. The maturation of egg boxes induced both a significant increase in their binding to wall-associated kinase 1 (WAK1) and an increased extracellular alkalinization when applied to Arabidopsis thaliana cell suspensions. The chemical modification of the reducing end of the OGAs largely diminished their elicitating activity but did not hinder either dimerization or binding of these end-reduced egg boxes to WAK1. We conclude that there are at least two different perception systems for egg box dimers. One binds egg box junctions and the other binds egg box ends. The relevance of these results is discussed in terms of pectic signal perception and plant-pathogen interaction. PMID- 18381976 TI - Capture of plasma membrane fragments from target cells by trogocytosis requires signaling in T cells but not in B cells. AB - Upon recognition of their respective cellular partners, T and B cells acquire their antigens by a process of membrane capture called trogocytosis. Here, we report that various inhibitors of actin polymerization or of kinases involved in intracellular signaling partially or fully inhibited trogocytosis by CD8(+) and CD4(+) T cells, whereas they had no effect on trogocytosis by B cells. Similarly, trogocytosis by T cells was inhibited at 4 degrees C, whereas in B cells it was independent of temperature, indicating that trogocytosis by B cells does not rely on active processes. By contrast, most inhibitors we tested impaired both T-cell and B-cell activation. The differential effect of inhibitors on T-cell and B-cell trogocytosis was not due to the higher affinity of the B-cell receptor for its cognate antigen compared with the affinity of the T-cell receptor for its own antigen, but it correlated tightly with the abilities of T cells and B cells to form conjugates with their target cells in the presence of inhibitors. Trogocytosis thus has different requirements in different cell types. Moreover, the capture of membrane antigen by B cells is identified as a novel signaling independent event of B-cell biology. PMID- 18381978 TI - Time for renewed commitment to viral hepatitis prevention. PMID- 18381979 TI - Bandoeng Conference of 1937. PMID- 18381980 TI - 2007 murder rates in New Orleans, Louisiana. PMID- 18381981 TI - VA disability policies and posttraumatic morbidity. PMID- 18381982 TI - Military-related PTSD, current disability policies, and malingering. PMID- 18381983 TI - The Sungari River flood and the Jewish community in Harbin, China. PMID- 18381984 TI - Who's using and who's doing time: incarceration, the war on drugs, and public health. PMID- 18381985 TI - Keeping guns out of the hands of abusers: handgun purchases and restraining orders. AB - Persons under certain domestic violence restraining orders are prohibited by federal law from purchasing and possessing a firearm. We used administrative data from California to link 794426 restraining orders with 1388724 handgun purchase applications. We found that restrained persons were not a less law-abiding group in general, but they appeared to be repeatedly or serially abusive to intimate partners, and their handgun purchase rates were highest after their restraining orders expired. PMID- 18381986 TI - Random drug testing in US public school districts. AB - We estimated the proportion of the nation's public school districts that have high school grades in which random drug testing is conducted. We collected data in spring 2005 from 1343 drug prevention coordinators in a nationally representative sample of school districts with schools that have high school grades; of these districts, 14% conducted random drug testing. Almost all districts randomly tested athletes, and 65% randomly tested other students engaged in extracurricular activities; 28% randomly tested all students, exceeding the current sanction of the US Supreme Court. PMID- 18381987 TI - Barriers to obtaining diagnostic testing for coronary artery disease among veterans. AB - OBJECTIVES: We sought to identify factors associated with appointment nonattendance for diagnostic testing of coronary artery disease among veterans. For patients with possible heart disease, appointment nonattendance may seriously compromise short- and long-term outcomes. Understanding factors associated with nonattendance may help improve care while reducing inefficiency in service delivery. METHODS: We surveyed patients who attended (n = 240) or did not attend (n = 139) a scheduled cardiac appointment at a midwestern Veterans Administration medical center. Multivariable regression models were used to assess factors associated with nonattendance. RESULTS: Younger age, lower income, unemployment, and longer wait times for appointments were predictive of nonattendance. Nonattenders reported fewer cardiac symptoms and were more likely to attribute their symptoms to something other than heart disease. Nonattendance was also associated with a coping style characterized by avoidance of aversive information. Logistical issues, fear of diagnostic procedures, disbelief that one had heart disease, and medical mistrust were some of the reasons given for missed appointments. CONCLUSIONS: Appointment nonattendance among veterans scheduled for cardiology evaluation was associated with several important cognitive factors. These factors should be considered when one is designing clinical systems to reduce patient nonattendance. PMID- 18381988 TI - Correlates of the stages of change for physical activity in a population survey. AB - OBJECTIVES: We sought to identify variables associated with being in a particular stage of change for physical activity-a measure of behavioral intention to engage in regular physical activity. Understanding behavioral intentions can be useful in explaining why individuals are physically inactive or active. METHODS: Data from the Rhode Island 2000 Behavioral Risk Factor Surveillance System were used to evaluate predictors of stage of change for physical activity. There were 3454 observations in the data set, representing a weighted population of 742636 people. Estimates were obtained from polytomous multiple logistic models. RESULTS: Being a woman, Hispanic, non-Hispanic Black, and older than 55 years of age were associated with being in precontemplation and contemplation stages of change rather than maintenance. Self-perceived health status and rarely feeling healthy or full of energy were strongly predictive of stage of change. Having a health limitation was a dichotomous predictor, predicting being in action and precontemplation stages. CONCLUSIONS: Several sociodemographic and health variables were associated with varying patterns of stages of change for physical activity. The complexity of individual intentions for physical activity provides evidence for the potential existence of mediating, effect-modifying, and confounding variables that differ depending on individual characteristics. PMID- 18381989 TI - New York City's initiatives on diabetes and HIV/AIDS: implications for patient care, public health, and medical professionalism. AB - Two recent New York City Department of Health and Mental Hygiene initiatives expanded the mission and scope of public health, with implications for both New York and the nation. The programs target diabetes and HIV/AIDS for greater systemic and expanded reporting, surveillance, and intervention. These initiatives do not balance heightened surveillance and intervention with the provision of meaningful safeguards or resources for prevention and treatment. The programs intrude on the doctor-patient relationship and may alienate the very patients and health professionals they aim to serve. Better models are available to achieve their intended goals. These initiatives should be reconsidered so that such an expansion of public health authority in New York City does not become part of a national trend. PMID- 18381990 TI - Characteristics of physically active smokers and implications for harm reduction. AB - OBJECTIVES: We sought to establish the prevalence of physical activity among smokers, whether or not physically active smokers were more likely to attempt cessation, and who these physically active smokers were. METHODS: We used logistic regression to contrast physically active and inactive smokers in a secondary data analysis of the Canadian Community Health Survey Cycle 1.1. RESULTS: Physically active smokers represented almost one quarter of the smoking population. Compared with physically inactive smokers, physically active smokers were more likely to have attempted cessation in the past year. Physically active smokers were more likely to be young, single, and men compared with their inactive counterparts. Income had no influence in distinguishing physically active and inactive smokers. CONCLUSIONS: Skepticism persists regarding the practicality and potential risks of promoting physical activity as a harm reduction strategy for tobacco use. We found that a modest proportion of the daily smoking population was physically active and that engagement in this behavior was related to greater cessation attempts. Interventions could be developed that target smokers who are likely to adopt physical activity. PMID- 18381991 TI - Intervening to reduce inequalities in infections in Europe. AB - The European Centre for Disease Prevention and Control was founded in response to newly emerging infections such as severe acute respiratory syndrome and avian influenza. However, Europe faces other communicable disease challenges that have proven to be remarkably resilient to public health interventions. We present examples of communicable diseases with inequitable distribution among those with poor educational attainment, low income, or other socioeconomic factors in every European country. Because these findings are incompatible with social justice and fairness, we examine strategic interventions targeting upstream causes of communicable disease transmission keeping in mind 10 indispensable public health functions essential to reach marginalized groups. These interventions have to be tailored to the socio-political context and rely on community-based decision making and intersectorial collaboration. PMID- 18381992 TI - Population attributable fractions of psychiatric disorders and suicide ideation and attempts associated with adverse childhood experiences. AB - OBJECTIVES: We sought to determine the fractions of psychiatric disorders and suicide ideation and attempts in a general population sample attributable to childhood physical abuse, sexual abuse, and witnessing domestic violence. METHODS: Data were obtained from the US National Comorbidity Survey Replication. Population attributable fractions were calculated to determine the proportion of psychiatric disorders and suicide ideation and attempts attributable to adverse childhood experiences. The analysis was stratified by gender. RESULTS: The estimated attributable fractions for psychiatric disorders attributable to having experienced any adverse childhood event ranged from 22% to 32% among women and 20% to 24% among men. Having experienced any adverse event accounted for a substantial proportion of suicide ideation and attempts among women (16% and 50%, respectively) and men (21% and 33%, respectively). Substantial proportions of poor mental health outcomes were also attributable to increasing number of adverse events. CONCLUSIONS: The estimated proportions of poor mental health outcomes attributed to childhood adversity were medium to large for men and women. Prevention efforts that reduce exposure to adverse childhood events could substantially reduce the prevalence of psychopathology and suicidal behavior in the general population. PMID- 18381993 TI - Trends in recorded influenza mortality: United States, 1900-2004. AB - OBJECTIVES: I sought to describe trends in historical influenza mortality data in the United States since 1900 and compare pandemic with nonpandemic influenza seasons. METHODS: I compiled a database of monthly influenza-classed death rates from official US mortality tables for the years 1900 to 2004 (1905-1909 excluded), from which I calculated adjusted influenza season (July 1-June 30) mortality rates. RESULTS: An overall and substantial decline in influenza-classed mortality was observed during the 20th century, from an average seasonal rate of 10.2 deaths per 100 000 population in the 1940s to 0.44 per 100 000 by the 1990s [corrected] . The 1918-1919 pandemic stands out as an exceptional outlier. The 1957-1958 and 1968-1969 influenza pandemic seasons, by contrast, displayed substantial overlap in both degree of mortality and timing compared with nonpandemic seasons. CONCLUSIONS: The considerable similarity in mortality seen in pandemic and non-pandemic influenza seasons challenges common beliefs about the severity of pandemic influenza. The historical decline in influenza-classed mortality rates suggests that public health and ecological factors may play a role in influenza mortality risk. Nevertheless, the actual number of influenza attributable deaths remains in doubt. PMID- 18381994 TI - Discharge planning and continuity of health care: findings from the San Francisco County Jail. AB - Continuity of health care among the formerly incarcerated is an emerging public health challenge. We used data from the San Francisco County Jail to determine whether discharge planning improves access to care on release. Inmates who were HIV positive and received discharge planning were 6 times more likely to have a regular source of care in the community compared with inmates with other chronic medical conditions, and they were as likely to have a regular source of care compared with the general San Francisco population. PMID- 18381995 TI - States and substance abuse treatment programs: funding and guidelines for infection-related services. AB - Community-based substance abuse treatment programs provide HIV, hepatitis C virus, and sexually transmitted infection services. To explore how state funding and guidelines affect practice, we surveyed state agency administrators and substance abuse treatment program administrators and clinicians regarding 8 infection-related services. Although state funding for infection-related services is widely available, substance abuse treatment programs do not always access it. Substance abuse treatment program guidelines are clearer in states that have written guidelines. Improved communication between state agencies and substance abuse treatment programs may enhance service. PMID- 18381996 TI - Status of influenza and pneumococcal vaccination among older American Indians and Alaska Natives. AB - OBJECTIVES: We sought to estimate the influenza and pneumococcal vaccination coverage among older American Indian and Alaska Native (AIAN) adults nationally and the impact of sociodemographic factors, variations by geographic region, and access to services on vaccination coverage. METHODS: We obtained our sample of 1981 AIAN and 179845 White respondents 65 years and older from Behavioral Risk Factor Surveillance System data from 2003 to 2005. Logistic regression provided predictive marginal vaccination coverage for each covariate and adjusted for demographic characteristics and access to care. RESULTS: Unadjusted influenza coverage estimates were similar between AIAN and White respondents (68.1% vs 69.5%), but pneumococcal vaccination was lower among AIAN respondents (58.1% vs 67.2%; P<.01). After multivariable adjustment for sociodemographic characteristics, self-reported coverage for both vaccines was statistically similar between AIAN and White adults. CONCLUSIONS: Although there was no disparity in influenza coverage, pneumococcal coverage was lower among AIAN than among White respondents, probably because of sociodemographic risk factors. Regional variation indicates a need to monitor coverage and target interventions to reduce disparities within geographically and culturally diverse subpopulations of AIAN persons. PMID- 18381997 TI - Health effects of exposure to water-damaged New Orleans homes six months after Hurricanes Katrina and Rita. AB - OBJECTIVES: We investigated the relation between respiratory symptoms and exposure to water-damaged homes and the effect of respirator use in posthurricane New Orleans, Louisiana. METHODS: We randomly selected 600 residential sites and then interviewed 1 adult per site. We created an exposure variable, calculated upper respiratory symptom (URS) and lower respiratory symptom (LRS) scores, and defined exacerbation categories by the effect on symptoms of being inside water damaged homes. We used multiple linear regression to model symptom scores (for all participants) and polytomous logistic regression to model exacerbation of symptoms when inside (for those participating in clean-up). RESULTS: Of 553 participants (response rate = 92%), 372 (68%) had participated in clean-up; 233 (63%) of these used a respirator. Respiratory symptom scores increased linearly with exposure (P < .05 for trend). Disposable-respirator use was associated with lower odds of exacerbation of moderate or severe symptoms inside water-damaged homes for URS (odds ratio (OR) = .51; 95% confidence interval (CI) = 0.24, 1.09) and LRS (OR = 0.33; 95% CI = 0.13, 0.83). CONCLUSIONS: Respiratory symptoms were positively associated with exposure to water-damaged homes, including exposure limited to being inside without participating in clean-up. Respirator use had a protective effect and should be considered when inside water-damaged homes regardless of activities undertaken. PMID- 18381998 TI - Smoking-cessation media campaigns and their effectiveness among socioeconomically advantaged and disadvantaged populations. AB - OBJECTIVES: We examined whether the impact of televised smoking cessation ads differed by a population's education and income. METHODS: We used longitudinal data from the Wisconsin Behavioral Health Survey, a statewide sample of 452 adult smokers who were interviewed in 2003 to 2004 and followed up 1 year later. Logistic regression was used to assess whether baseline recall of secondhand smoke ads and "keep trying to quit" ads was associated with quit attempts and smoking abstinence at 1 year. Interaction terms were used to assess whether these associations differed by the smokers' education and income levels. RESULTS: Overall, neither keep-trying-to-quit nor secondhand smoke ad recall was associated with quit attempts or smoking abstinence. Keep-trying-to-quit ads were significantly more effective in promoting quit attempts among higher-versus lower educated populations. No differences were observed for secondhand smoke ads by the smokers' education or income levels. CONCLUSIONS: Some media campaign messages appear less effective in promoting quit attempts among less-educated populations compared with those who have more education. There is a need to develop media campaigns that are more effective with less-educated smokers. PMID- 18381999 TI - On wit, irony, and living with imperfection: how Britain said no to abstinence. AB - Christian perfection, the evangelical doctrine that gave rise to abstinence as it is understood and practiced in America, originated in Britain with John Wesley and the Methodists. We examine why that doctrine floundered in its country of origin, opening the door to a more pluralistic and evidence-based approach to problems such as alcohol and drug abuse. Although social and political factors were important (the stratification of British society stood in the way of holding everyone to the same moral standard, and the drink trade was far better organized than its American counterpart), Britain's intellectual elite also played a vital role, heaping ridicule on the temperance movement and subjecting it to a devastating critique. PMID- 18382000 TI - The effect of vaccinated children on increased hepatitis B immunization among high-risk adults. AB - OBJECTIVES: We sought to examine trends in hepatitis B virus (HBV) vaccination among high-risk adults and assess the potential effect vaccinated adolescents have on these trends as they age. METHODS: We used data from the National Health Interview Survey 2000, 2002, and 2004 to examine trends in HBV vaccination among high-risk adults aged 18 to 49 years and in age subgroups (18-29, 30-39, and 40 49 years). We investigated temporal differences in vaccination rates for the 18- to 29-year-old cohort with model-based linear contrasts constructed from a logistic regression model with age and survey year as predictors. RESULTS: There was a significant increasing trend in vaccination prevalence across the 3 survey years (32.6%, 35.3%, and 41.4%; trend test, P=.001). We found that respondents aged 18 to 29 years were more likely to be vaccinated in 2004 than in 2000, after adjusting for relevant confounders (odds ratio=1.73; 95% confidence interval=1.14, 2.6); there was no significant increase in vaccination for the other cohorts. CONCLUSIONS: A cohort effect, in which successfully vaccinated adolescents have reached young adulthood, contributes significantly to recent trends showing improved HBV vaccination among high-risk adults. PMID- 18382001 TI - Long-term trends in adolescent and young adult smoking in the United States: metapatterns and implications. AB - OBJECTIVES: We sought to describe long-term adolescent and young adult smoking trends and patterns. METHODS: We analyzed adolescent data from Monitoring the Future, 1976 to 2005, and young adult (aged 18-24 years) data from the National Health Interview Survey, 1974 to 2005, overall and in subpopulations to identify trends in current cigarette smoking prevalence. RESULTS: Five metapatterns emerged: we found (1) a large increase and subsequent decrease in overall smoking over the past 15 years, (2) a steep decline in smoking among Blacks through the early 1990s, (3) a gender gap reversal among older adolescents and young adults who smoked over the past 15 years, (4) similar trends in smoking for most subgroups since the early 1990s, and (5) a large decline in smoking among young adults with less than a high school education. CONCLUSIONS: Long-term patterns for adolescent and young adult cigarette smoking were decidedly nonlinear, and we found evidence of a cohort effect among young adults. Continued strong efforts and a long-term societal commitment to tobacco use prevention are needed, given the unprecedented declines in smoking among most subpopulations since the mid- to late 1990s. PMID- 18382002 TI - The relationship between in-home water service and the risk of respiratory tract, skin, and gastrointestinal tract infections among rural Alaska natives. AB - OBJECTIVES: We investigated the relationship between the presence of in-home piped water and wastewater services and hospitalization rates for respiratory tract, skin, and gastrointestinal tract infections in rural Alaska. METHODS: We determined in-home water service and hospitalizations for selected infectious diseases among Alaska Natives by region during 2000 to 2004. Within 1 region, infant respiratory hospitalizations and skin infections for all ages were compared by village-level water services. RESULTS: Regions with a lower proportion of home water service had significantly higher hospitalization rates for pneumonia and influenza (rate ratio [RR] = 2.5), skin or soft tissue infection (RR = 1.9), and respiratory syncytial virus (RR = 3.4 among those younger than 5 years) than did higher-service regions. Within 1 region, infants from villages with less than 10% of homes served had higher hospitalization rates for pneumonia (RR = 1.3) and respiratory syncytial virus (RR = 1.2) than did infants from villages with more than 80% served. Outpatient Staphylococcus aureus infections (RR = 5.1, all ages) and skin infection hospitalizations (RR = 2.7, all ages) were higher in low-service than in high-service villages. CONCLUSIONS: Higher respiratory and skin infection rates were associated with a lack of in home water service. This disparity should be addressed through sanitation infrastructure improvements. PMID- 18382003 TI - Being poor and coping with stress: health behaviors and the risk of death. AB - OBJECTIVES: Individuals may cope with perceived stress through unhealthy but often pleasurable behaviors. We examined whether smoking, alcohol use, and physical inactivity moderate the relationship between perceived stress and the risk of death in the US population as a whole and across socioeconomic strata. METHODS: Data were derived from the 1990 National Health Interview Survey's Health Promotion and Disease Prevention Supplement, which involved a representative sample of the adult US population (n = 40335) and was linked to prospective National Death Index mortality data through 1997. Gompertz hazard models were used to estimate the risk of death. RESULTS: High baseline levels of former smoking and physical inactivity increased the impact of stress on mortality in the general population as well as among those of low socioeconomic status (SES), but not middle or high SES. CONCLUSIONS: The combination of high stress levels and high levels of former smoking or physical inactivity is especially harmful among low-SES individuals. Stress, unhealthy behaviors, and low SES independently increase risk of death, and they combine to create a truly disadvantaged segment of the population. PMID- 18382004 TI - Reduction of state victim compensation disparities in disadvantaged crime victims through active outreach and assistance: a randomized trial. AB - OBJECTIVES: We examined whether providing active outreach and assistance to crime victims as part of comprehensive psychosocial services reduced disparities in access to state compensation funds. METHODS: We analyzed data from a randomized trial of injured crime victims (N = 541) and compared outcomes from comprehensive psychosocial services with usual community care. We examined the impact of outreach and assistance on disparities in applying for victim compensation by testing for interactions between victim characteristics and treatment condition in logistic regression analyses. RESULTS: Victims receiving comprehensive services were much more likely to apply for victim compensation than were victims receiving usual care. Comprehensive services decreased disparities associated with younger age, lower levels of education, and homelessness. CONCLUSIONS: State level victim compensation funds are available to help individuals recover physically, psychologically, and financially from crime victimization. However, few crime victims apply for victim compensation, and there are particularly low application rates among young, male, ethnic minority, and physical assault victims. Active outreach and assistance can address disparities in access to victim compensation funds for disadvantaged populations and should be offered more widely to victims of violent crime. PMID- 18382006 TI - Bioterrorism surveillance and privacy: intersection of HIPAA, the Common Rule, and public health law. AB - The threat of bioterrorism in the wake of the September 11, 2001, terrorist attacks cannot be ignored. Syndromic surveillance, the practice of electronically monitoring and reporting real-time medical data to proactively identify unusual disease patterns, highlights the conflict between safeguarding public health while protecting individual privacy. Both the Health Insurance Portability and Accountability Act and the Common Rule (which promulgates protections for individuals in federally sponsored medical research programs) safeguard individuals. Public health law protects the entire populace; uneven state-level implementation lacks adequate privacy protections. We propose 3 models for a nationwide bioterrorism surveillance review process: a nationally coordinated systems approach to using protected health information, creating public health information privacy boards, expanding institutional review boards, or some combination of these. PMID- 18382005 TI - A randomized intervention trial to reduce the lending of used injection equipment among injection drug users infected with hepatitis C. AB - OBJECTIVES: We evaluated the efficacy of a peer-mentoring behavioral intervention designed to reduce risky distributive injection practices (e.g., syringe lending, unsafe drug preparation) among injection drug users with hepatitis C virus (HCV) infection. METHODS: A randomized trial with a time-equivalent attention-control group was conducted among 418 HCV-positive injection drug users aged 18 to 35 years in 3 US cities. Participants reported their injection-related behaviors at baseline and at 3- and 6-month follow-ups. RESULTS: Compared with the control group, intervention-group participants were less likely to report distributive risk behaviors at 3 months (odds ratio [OR]=0.46; 95% confidence interval [CI]=0.27, 0.79) and 6 months (OR=0.51; 95% CI=0.31, 0.83), a 26% relative risk reduction, but were no more likely to cite their HCV-positive status as a reason for refraining from syringe lending. Effects were strongest among intervention group participants who had known their HCV-positive status for at least 6 months. Peer mentoring and self-efficacy were significantly increased among intervention group participants, and intervention effects were mediated through improved self efficacy. CONCLUSIONS: This behavioral intervention reduced unsafe injection practices that may propagate HCV among injection drug users. PMID- 18382007 TI - Racial differences in the evaluation and treatment of hepatitis C among veterans: a retrospective cohort study. AB - OBJECTIVES: We examined the association between race and hepatitis C virus (HCV) evaluation and treatment of veterans in the Northwest Network of the Department of Veterans Affairs (VA). METHODS: In our retrospective cohort study, we used medical records to determine antiviral treatment of 4263 HCV-infected patients from 8 VA medical centers. Secondary outcomes included specialty referrals, laboratory evaluation, viral genotype testing, and liver biopsy. Multiple logistic regression was used to adjust for clinical (measured through laboratory results and International Classification of Diseases, Ninth Revision, codes) and sociodemographic factors. RESULTS: Blacks were less than half as likely as Whites to receive antiviral treatment (odds ratio [OR]=0.38; 95% confidence interval [CI]=0.23, 0.63). Both had similar odds of referral and liver biopsy. However, Blacks were significantly less likely to have complete laboratory evaluation (OR=0.67; 95% CI=0.52, 0.88) and viral genotype testing (OR=0.68; 95% CI=0.51, 0.90). CONCLUSIONS: Race is associated with receipt of medical care for various medical conditions. Further investigation is warranted to help understand whether patient preference or provider bias may explain why HCV-infected Blacks were less likely to receive medical care than Whites. PMID- 18382008 TI - Effects of enrollment in medicaid versus the state children's health insurance program on kindergarten children's untreated dental caries. AB - OBJECTIVES: We compared levels of untreated dental caries in children enrolled in public insurance programs with those in nonenrolled children to determine the impact of public dental insurance and the type of plan (Medicaid vs State Children's Health Insurance Program [SCHIP]) on untreated dental caries in children. METHODS: Dental health outcomes were obtained through a calibrated oral screening of kindergarten children (enrolled in the 2000-2001 school year). We obtained eligibility and claims data for children enrolled in Medicaid and SCHIP who were eligible for dental services during 1999 to 2000. We developed logistic regression models to compare children's likelihood and extent of untreated dental caries according to enrollment. RESULTS: Children enrolled in Medicaid or SCHIP were 1.7 times (95% confidence interval [CI] = 1.65, 1.77) more likely to have untreated dental caries than were nonenrolled children. SCHIP-enrolled children were significantly less likely to have untreated dental caries than were Medicaid enrolled children (odds ratio [OR]=0.74; 95% CI=0.67, 0.82). According to a 2 part regression model, children enrolled in Medicaid or SCHIP have 17% more untreated dental caries than do nonenrolled children, whereas those in SCHIP had 16% fewer untreated dental caries than did those in Medicaid. CONCLUSIONS: Untreated tooth decay continues to be a significant problem for children with public insurance coverage. Children who participated in a separate SCHIP program had fewer untreated dental caries than did children enrolled in Medicaid. PMID- 18382010 TI - Ethics in public health research: privacy and public health at risk: public health confidentiality in the digital age. AB - Public health agencies increasingly use electronic means to acquire, use, maintain, and store personal health information. Electronic data formats can improve performance of core public health functions, but potentially threaten privacy because they can be easily duplicated and transmitted to unauthorized people. Although such security breaches do occur, electronic data can be better secured than paper records, because authentication, authorization, auditing, and accountability can be facilitated. Public health professionals should collaborate with law and information technology colleagues to assess possible threats, implement updated policies, train staff, and develop preventive engineering measures to protect information. Tightened physical and electronic controls can prevent misuse of data, minimize the risk of security breaches, and help maintain the reputation and integrity of public health agencies. PMID- 18382009 TI - The influence of perceived risk to health and immigration-related characteristics on substance use among Latino and other immigrants. AB - OBJECTIVES: We examined whether immigration-related characteristics and perceptions of risk surrounding substance use were independently associated with lifetime use of cigarettes and various illicit substances among immigrant and native-born Latino and non-Latino White adults in the United States. METHODS: Data were from the 2002 National Survey on Drug Use and Health. Analyses were limited to Latinos and non-Latino Whites 18 years and older. We used cross tabulations and multivariate logistic regression to test relations between risk perceptions, immigration characteristics, and substance use. RESULTS: More than two thirds of all respondents perceived moderate or great risk to health and well being associated with all substances analyzed. The odds of lifetime substance use by Latino and non-Latino White immigrants were lower than for US-born non-Latino Whites. Immigrant Latinos' odds of lifetime substance use were lower than for US born Latinos. Moderate or great perceived risk was associated with lower likelihood of lifetime use of all substances except cigarettes. CONCLUSIONS: Foreign birth appeared to protect against substance use among both Latino and non Latino White immigrants. Future studies should examine potential protective factors, including cultural beliefs and practices, acculturation, familial ties, and social network influences. PMID- 18382011 TI - Communicating with the public about emerging health threats: lessons from the Pre Event Message Development Project. AB - OBJECTIVES: We sought to better understand the challenges of communicating with the public about emerging health threats, particularly threats involving toxic chemicals, biological agents, and radioactive materials. METHODS: At the request of the Centers for Disease Control and Prevention, we formed an interdisciplinary consortium of investigative teams from 4 schools of public health. Over 2 years, the investigative teams conducted 79 focus group interviews with 884 participants and individual cognitive response interviews with 129 respondents, for a total sample of 1013 individuals. The investigative teams systematically compared their results with other published research in public health, risk communication, and emergency preparedness. RESULTS: We found limited public understanding of emerging biological, chemical, and radioactive materials threats and of the differences between them; demand for concrete, accurate, and consistent information about actions needed for protection of self and family; active information seeking from media, local authorities, and selected national sources; and areas in which current emergency messaging can be improved. CONCLUSIONS: The public will respond to a threat situation by seeking protective information and taking self-protective action, underlining the critical role of effective communication in public health emergencies. PMID- 18382013 TI - Penicilliosis: an AIDS-defining disease in Asia. PMID- 18382012 TI - The Young Men's Survey phase II: hepatitis B immunization and infection among young men who have sex with men. AB - OBJECTIVES: We measured the prevalence of hepatitis B virus (HBV) immunization and HBV infection among men aged 23 to 29 years who have sex with men. METHODS: We analyzed data from 2834 men who have sex with men in 6 US metropolitan areas. Participants were interviewed and tested for serologic markers of immunization and HBV infection in 1998 through 2000. RESULTS: Immunization prevalence was 17.2%; coverage was 21.0% among participants with private physicians or health maintenance organizations and 12.6% among those with no source of health care. Overall, 20.6% had markers of HBV infection, ranging from 13.7% among the youngest to 31.0% among the oldest participants. Among those susceptible to HBV, 93.5% had regular sources of health care, had been tested for HIV, or had been treated for a sexually transmitted disease. CONCLUSIONS: Although many young men who have sex with men have access to health care, most are not immunized against HBV. To reduce morbidity from HBV in this population, providers of health care, including sexually transmitted disease and HIV prevention services, should provide vaccinations or referrals for vaccination. PMID- 18382014 TI - Magnetic resonance whole body imaging at 3 Tesla: feasibility and findings in a cohort of asymptomatic medical doctors. AB - OBJECTIVES: To evaluate the feasibility of whole body imaging using a 3 Tesla magnetic resonance scanner without a contrast agent, and to study the prevalence of abnormal findings among a cohort of asymptomatic doctors. DESIGN: Prospective study. SETTING: Private hospital, Hong Kong. PARTICIPANTS: A total of 132 asymptomatic medical doctors (111 men, 21 women), with a mean age of 56 (range, 38-82) years, volunteered for the study. They underwent corresponding whole body imaging at our hospital between October 2005 and February 2006. Imaging involved a 3 Tesla magnetic resonance scanner with 32 channels, parallel imaging, Total Imaging Matrix technology, a maximum gradient amplitude of 40 mT/m and a slew rate of 200 mT/m/ms (Magnetom Tim Trio, Siemens Medical Solution, Erlangen, Germany). The use of matrix coils enabled coverage of the whole body. No contrast agent was used. MAIN OUTCOME MEASURES: Detection of abnormalities in asymptomatic, apparently healthy adults. RESULTS: All examinations were completed successfully. The mean scan time per subject was 33 (standard deviation, 4) minutes. All subjects tolerated the examination well and overall imaging quality was satisfactory. A total of 124 (94%) subjects had positive findings, of whom 24 (18%) had further workup. Five (4%) subjects were found to have tumours, of which two (1.5%) were proven malignant. Our cancer detection rate was comparable to that of other reported whole body screening studies using contrast magnetic resonance imaging and positron emission tomography. CONCLUSION: We demonstrated the feasibility of performing whole body imaging in 30 minutes, using 32-channel magnetic resonance imaging at 3 Tesla without a contrast agent or any ionising radiation. PMID- 18382015 TI - Clinical profile of young children with mental retardation and developmental delay in Hong Kong. AB - OBJECTIVE: To report the clinical profile of children with mental retardation and developmental delay diagnosed by the Child Assessment Service. DESIGN: Retrospective study. SETTING: Child Assessment Service, Department of Health, Hong Kong. PARTICIPANTS: Data pertaining to the children with mental retardation and developmental delay were drawn from an in-house clinical information system in the year 2004. MAIN OUTCOME MEASURES: Clinical profiles including: sources, reasons and age of referral, diagnosis, gender ratio, co-morbidities, and socio economic background. RESULTS: In 2004, 23% (1463 of 6439) of Child Assessment Service referrals were diagnosed to have mental retardation or developmental delay. The Family Health Service was the major source of referral (64%). The majority (93%) of children were referred before the age of 6 years. The most common reason for referral was language delay (39%). More boys were affected (3 boys: 1 girl). The two most common co-morbidities were autistic spectrum disorders (33% in mental retardation and 19% in developmental delay) and discrepant language delay (17% in mental retardation and 47% in developmental delay). The socio-economic status of these families was higher than those in the general population. CONCLUSION: The data presented here provide information on the descriptive epidemiology of mental retardation and developmental delay among Hong Kong children. Since mental retardation and developmental delay are common developmental disabilities in Hong Kong, public health education to promote and ensure early screening and identification of cases is an important prelude to early training and guidance for families with children having these conditions. PMID- 18382016 TI - Clinical presentations and outcomes of Penicillium marneffei infections: a series from 1994 to 2004. AB - OBJECTIVES: To describe the clinical presentation, management, and outcomes of patients with Penicillium marneffei infections in a regional hospital in Hong Kong. DESIGN: Retrospective study. SETTING: A regional and tertiary human immunodeficiency virus-referral hospital in Hong Kong. PATIENTS: Those who had penicilliosis during the inclusive period January 1994 to February 2004. RESULTS: Forty-seven immunocompromised patients (44 being human immunodeficiency virus positive) with penicilliosis were retrospectively studied. Fever, malaise, and anaemia were the commonest presentations. Most diagnoses were obtained from blood cultures (83%) and lymph node biopsies (34%). Five (11%) died, death being attributable to penicilliosis; four (9%) of them had received no specific antifungal treatment due to late presentation and late diagnosis. The CD4 count of human immunodeficiency virus-infected patients upon diagnosis of penicilliosis was low (median, 20.0 cells/mm3). Most (70%) patients received amphotericin B as an induction treatment, followed by oral itraconazole, although a smaller proportion (21%) received oral itraconazole only. All surviving human immunodeficiency virus-infected patients took highly active antiretroviral treatment and oral itraconazole as secondary prophylaxis after treatment of penicilliosis. The prognosis appeared satisfactory with early diagnosis and administration of appropriate antifungal therapy. Relapse ensued in two (4%) of the patients only. CONCLUSION: Penicillium marneffei infection in immunocompromised patients is a serious disease with significant mortality if not diagnosed early and treated with appropriate antifungal drugs. Simple investigations like blood culture enable the diagnosis in the majority of cases. Immunocompromised patients who have been successfully treated should receive oral itraconazole as a maintenance therapy to prevent relapse. PMID- 18382017 TI - Personal use and professional recommendations of complementary and alternative medicine by Hong Kong registered nurses. AB - OBJECTIVE: To provide an understanding of Hong Kong registered nurses' personal and professional use of complementary and alternative medicine. DESIGN: Cross sectional questionnaire study. PARTICIPANTS: Registered nurses who were members of the Hong Kong College of Nursing were invited to participate. MAIN OUTCOME MEASURES: Demographic data of the respondents, prevalence of personal and professional use of complementary and alternative medicines, including their use for detailed clinical conditions. RESULTS: A total of 187 nurses participated in this study. Nearly four fifths (80%; 95% confidence interval, 74-86%) of the participants had used at least one form of complementary/alternative medicine. In addition to the personal use of such treatment, over two fifths (41%; 95% confidence interval, 34-48%) had recommended at least one form of complementary/alternative medicine to their patients. These included bone-setting (20%), Chinese remedial massage (tuina, 19%), and meditation (19%). Specifically, registered nurses recommended acupuncture or acupressure to patients with musculoskeletal disorders, chronic pain, or headaches/migraines. CONCLUSIONS: Personal use and professional recommendations for complementary and alternative medicine by registered nurses in Hong Kong is substantial. Registered nurses played an active role in advising such treatment for their patients based on their personal knowledge of perceived benefit in specific conditions. Further investigations with a larger sample size should focus on registered nurses' educational needs in respect of complementary and alternative medicine and the advice they recommended. PMID- 18382018 TI - Treatment outcome of rhabdomyosarcoma in Hong Kong Chinese children. AB - OBJECTIVES: To review the treatment outcome of rhabdomyosarcoma in Hong Kong Chinese children. DESIGN: Retrospective review. SETTING: University teaching hospital, Hong Kong. PATIENTS: Consecutive cases of rhabdomyosarcoma diagnosed and treated by the Department of Paediatrics and Adolescent Medicine of Queen Mary Hospital between 1989 and 2005. Each patient was staged and treated according to the Intergroup Rhabdomyosarcoma Study guidelines. MAIN OUTCOME MEASURES: Overall and event-free survival rates, and toxicity data. RESULTS: Of 19 patients (8 males and 11 females), 14 (74%) were younger than 10 years old. The median age at diagnosis was 6 (range, 0.5-17) years. Primary sites of rhabdomyosarcoma included: the head and neck (n=8; 6 classified as cranial parameningeal), genitourinary (3), extremity (3), pelvis (3), and trunk (2). Thirteen (68%) had embryonal and six (32%) had alveolar histology. Two, 2, 9, and 6 were classified as belonging to Intergroup Rhabdomyosarcoma Study groups 1, 2, 3, and 4, respectively. Respective 5-year overall and event-free survival rates of the entire cohort were 49% (95% confidence interval, 26-73%) and 32% (10-55%), with a median follow-up of 3.4 (range, 0.2-16.7) years. In non-metastatic cases (Intergroup Rhabdomyosarcoma Study groups 1-3), the 5-year overall survival rate was 66% (95% confidence interval, 39-93%) and in metastatic cases (group 4) it was 17% (0-46%). The 5-year overall survival rate for patients aged less than 10 years was 60% (95% confidence interval, 33-87%) compared to 20% (0-55%) in those aged 10 years and over. Significant treatment-related toxicities including myelosuppression, infections, peripheral neuropathy, and second cancers were encountered. CONCLUSIONS: Treatment outcome of rhabdomyosarcoma in this cohort of Chinese children was less favourable than that reported in international studies. Whilst the main reason could have been related to the high proportion of metastatic cases, also non-metastatic cases faired worse. Improved outcomes may be achieved by advances in multidisciplinary (paediatric oncology, pathology, radiotherapy, and surgery) management and supportive care. PMID- 18382019 TI - Role of Mycoplasma genitalium and Ureaplasma urealyticum in non-gonococcal urethritis in Hong Kong. AB - OBJECTIVE: To determine the association of Mycoplasma genitalium and Ureaplasma urealyticum in symptomatic male patients presenting with non-gonococcal urethritis in a sexually transmitted infection clinic in Hong Kong. DESIGN: Cross sectional study. SETTING: A sexually transmitted infection clinic, Department of Health, Centre for Health Protection, Hong Kong. PATIENTS: A cohort of consecutive new male patients attending the government sexually transmitted infection clinic. MAIN OUTCOME MEASURES: Prevalence of Mycoplasma genitalium and Ureaplasma urealyticum among symptomatic male patients with non-gonococcal urethritis and asymptomatic patients without non-gonococcal urethritis. RESULTS: Specimens of 22 and 10 patients tested positive by polymerase chain reaction for Ureaplasma urealyticum and Mycoplasma genitalium respectively, among the symptomatic non-gonococcal urethritis group (n=98). In the asymptomatic control group (n=236), corresponding patient numbers whose specimens tested positive were 47 and 5. There was no statistically significant difference between the two groups, in terms of the proportion of patients infected with Mycoplasma genitalium (P=0.799) or Ureaplasma urealyticum (P=0.535). CONCLUSIONS: In our study, demonstration of Mycoplasma genitalium and Ureaplasma urealyticum by polymerase chain reaction was not associated with symptomatic non-gonococcal urethritis in male patients attending a Hong Kong government clinic for sexually transmitted infections. PMID- 18382020 TI - HER2 overexpression of breast cancers in Hong Kong: prevalence and concordance between immunohistochemistry and in-situ hybridisation assays. AB - OBJECTIVES: To evaluate the prevalence of human epidermal growth factor receptor 2 (HER2) gene overexpression in breast cancer patients encountered in Hong Kong and the concordance of HER2 findings from primary immunohistochemistry assays and confirmatory in-situ hybridisation assays. DESIGN: Retrospective study. SETTING: Department of Clinical Oncology in a public hospital in Hong Kong. PATIENTS: All patient referrals between July 2006 and June 2007 with newly diagnosed invasive breast cancer (for prevalence evaluation), and all patients treated at our unit with confirmatory in-situ hybridisation tests performed within the study period (for concordance evaluation). RESULTS: There were 272 consecutive breast cancer patients eligible for prevalence evaluation. The distribution for immunohistochemistry staining in 249 cases for scores 0, 1+, 2+, and 3+ were 99 (40%), 40 (16%), 58 (23%), and 52 (21%) respectively. In the remaining 23 patients, four and 19 breast cancers were unscored and reported by immunohistochemistry to be HER2-positive and -negative, respectively. The overall HER2 overexpression rate (3+ or reported as positive) was 21%. HER2 overexpression was associated with grade 3 histology (P<0.001) and negative hormonal receptor status (P<0.001). However, it was not associated with age (P=0.525), T-classification (P=0.740), N-classification (P=0.691), nor group stages (P=0.433). Of the 37 patients with confirmatory in-situ hybridisation tests performed, 10 (71%) of 14 with immunohistochemistry staining of 3+ and 1 (4%) of 23 with immunohistochemistry staining of 2+ were found to have HER2 gene amplification. CONCLUSIONS: More than 25% of HER2 overexpression identified by immunohistochemistry assays in this Hong Kong cohort could not be verified by confirmatory in-situ hybridisation assays. Compliance with the latest guidelines for HER2 testing should improve the future accuracy and concordance. PMID- 18382021 TI - Gadolinium-enhanced magnetic resonance angiography in renal artery stenosis: comparison with digital subtraction angiography. AB - OBJECTIVES: To evaluate the accuracy of gadolinium-enhanced magnetic resonance angiography in assessing renal artery stenosis compared to catheter digital subtraction angiography. DESIGN: Retrospective study. SETTING: Singapore General Hospital. PATIENTS: Records of patients who underwent magnetic resonance angiography as well as digital subtraction angiography for assessment of renal artery stenosis from January 2003 to December 2005 were reviewed. RESULTS: There were 27 patients (14 male, 13 female) with a mean age of 62 (range, 44-77) years. There were 10 patients with renal transplants; their native renal arteries were not evaluated. Each of the two experienced interventional and body magnetic resonance radiologists, who were blinded to the results, reviewed the digital subtraction angiography and magnetic resonance angiography images respectively. Digital subtraction angiography was used as the standard of reference. A total of 39 renal arteries from these 27 patients were evaluated. One of the arteries was previously stented and could not be assessed with magnetic resonance angiography due to severe artefacts. Of the remaining 38 renal arteries, two were graded as normal, seven as having mild stenosis (<50%), eight as having moderate stenosis (> or =50% but <75%), and 21 as having severe stenosis (> or =75%). Magnetic resonance angiography and digital subtraction angiography were concordant in 89% of the arteries; magnetic resonance angiography overestimated the degree of stenosis in 8% and underestimated it in 3% of them. In the evaluation of clinically significant renal artery stenosis (> or =50%) with magnetic resonance angiography, the overall sensitivity, specificity, positive predictive value, and negative predictive value were 97%, 67%, 90%, and 86% respectively. The sensitivity and specificity of magnetic resonance angiography in transplant renal artery stenosis was 100%. CONCLUSION. Our experience suggested that gadolinium enhanced magnetic resonance angiography is a sensitive non-invasive modality useful in the assessment of clinically significant renal artery stenosis. PMID- 18382022 TI - Good's syndrome in a patient with cytomegalovirus retinitis. AB - Thymoma-related adult-onset immunodeficiency or Good's syndrome is an uncommon condition. This case, of a 50-year-old woman who was human immunodeficiency virus negative and developed herpes zoster and severe cytomegalovirus retinitis 6 months after removal of a thymoma, is the first to be reported in Hong Kong. Immunological investigations revealed no B cells, hypogammaglobulinaemia, a low CD4 count, and a low CD4/CD8 ratio. We recommend that immunological investigations, including T-cell subsets, B cells, and quantitative immunoglobulins, should be part of the routine diagnostic evaluation of patients with thymoma and infections. PMID- 18382023 TI - Splenic laceration after endoscopic retrograde cholangiopancreatography. AB - Endoscopic retrograde cholangiopancreatography has been part of clinical practice for over 35 years. The procedure itself carries risks. Most complications associated with the procedure have been described in the literature. Splenic injury is an extremely rare complication after endoscopic retrograde cholangiopancreatography and only nine such cases have been reported to our knowledge since 1988. We report on a patient who presented with abdominal pain and was found to have abnormal liver function and a dilated pancreatic duct. An endoscopic retrograde cholangiopancreatography was performed and a splenic laceration was noted subsequently. As this operation is an increasingly important diagnostic and therapeutic modality for pancreatico-biliary disease, clinicians must be aware of this complication in order to make an early diagnosis and begin appropriate management. PMID- 18382024 TI - Umbilical cord ulceration causing foetal haemorrhage and stillbirth. AB - We report a case of umbilical cord ulceration associated with obstruction of the duodeno-jejunal junction by a peritoneal band. Umbilical cord ulceration is a rare condition; a literature review identified a total of 17 cases only. In all cases, the ulceration was associated with congenital intestinal obstruction. Cord ulceration usually presents as sudden foetal deterioration due to foetal haemorrhage. This condition is associated with high perinatal mortality and morbidity. The causes of this condition are still unknown, and prenatal diagnosis is difficult. Awareness of the possible association between umbilical cord ulceration and intestinal obstruction, and of the need to deliver such pregnancies immediately when an abnormal foetal heart rate pattern develops might be the only means of preventing intrauterine death and improving neonatal outcomes. PMID- 18382025 TI - Caecal metastasis from a primary small-cell lung carcinoma. AB - Small bowel metastases from a primary lung carcinoma are rare. We report a case of a 59-year-old male with a primary small-cell lung carcinoma who developed anaemia and bowel symptoms. On colonoscopic examination he was found to have a tumour in the caecum near the ileocaecal valve, which was biopsied, revealing small neuroendocrine tumour cells. The patient then underwent systemic chemotherapy, which achieved a reduction in the size of the primary lung tumour and an improvement in his bowel symptoms. It is important that such a rare condition be recognised early as complicated intestinal metastases from a lung carcinoma can lead to high mortality rates and poor short-term outcome. With advances in chemotherapy and palliative care, patients with metastatic lung carcinoma can sometimes survive more than a year with reasonable quality of life. PMID- 18382026 TI - Brodie's abscess of the ulna caused by Salmonella typhi. AB - Osteomyelitis caused by Salmonella typhi is rare in patients with no haemoglobinopathies or other diseases causing immunosuppression. Brodie's abscess is a special variety of subacute or chronic osteomyelitis. An otherwise healthy woman who presented with forearm swelling for 6 months was diagnosed with a Brodie's abscess of the ulna caused by Salmonella typhi. Magnetic resonance imaging and a computed tomography-guided needle biopsy were performed. She was later found to be a Salmonella carrier. The Brodie's abscess was treated by surgical debridement and a course of antibiotics. The clinical, radiological, and management aspects of the disease are discussed. PMID- 18382027 TI - Arteriovenous malformation related to the pinna. AB - Arteriovenous malformations are rare in the head and neck region and generally arise from intracranial vessels. We present two rare cases with spontaneous arteriovenous malformations related to the ear. The role of magnetic resonance imaging and colour Doppler sonography in the diagnosis and management of such cases is discussed along with a review of the literature. PMID- 18382028 TI - A 50-year-old man with a submental mass. PMID- 18382029 TI - Bronchial artery embolisation in the management of chronic recurrent haemoptysis. PMID- 18382030 TI - Changes in the percentage of students who walk or bike to school-United States, 1969 and 2001. AB - BACKGROUND: This report describes changes in the percentage of US students (age 5 to 18 years) who walked or bicycled to school and in the distance that they lived from or traveled to their school in 1969 and 2001 and travel patterns in 2001. METHODS: Data were from the 1969 National Personal Transportation Survey report on school travel and the 2001 National Household Transportation Survey. RESULTS: A smaller percentage of students lived within 1 mile of school in 2001 than in 1969. The percentage of students who walked or biked any distance decreased from 42.0% to 16.2%. Nearly half of students used more than 1 travel mode or went to an additional destination en route between home and school in 2001. CONCLUSION: Multidisciplinary efforts are needed to increase the percentage of students who walk or bike to school, as well as decrease the distances that students travel. PMID- 18382031 TI - Walking the dog: is pet ownership associated with physical activity in California? AB - BACKGROUND: Several studies have reported positive associations between pet ownership and a variety of health outcomes. In this study, we explored associations between pet ownership and physical activity in a large, ethnically diverse population-based sample in California. METHOD: Data from the California Health Interview Survey (CHIS) were used to assess the associations between pet ownership (ie, dog, dog and cat, cat, and non-pet owners) and transportation and leisure walking in a sample of 41,514 adults. Logistic regression was used to assess associations between pet ownership and type of walking, and linear regression was used to assess associations between pet ownership and total minutes walking per week. RESULTS: Dog owners were slightly less likely to walk for transportation than were non-pet owners (OR = 0.91; 95% CI: 0.85 to 0.99) but more likely to walk for leisure than non-pet owners (OR = 1.6; 95% CI: 1.5 to 1.8) in multivariate analyses. Overall, dog owners walked 18.9 (95% CI: 11.4 to 26.4) minutes more per week than non-pet owners. Walking behaviors of cat owners were similar to non-pet owners. CONCLUSION: Our findings support the moderate association between dog ownership and higher levels of physical activity. PMID- 18382032 TI - Interrelations of socioeconomic position and occupational and leisure-time physical activity in the National Health and Nutrition Examination Survey. AB - BACKGROUND: The interrelations between various physical activity domains have received little empirical attention in the United States. Of particular interest, given the potential applicability to traditionally underserved communities, is the nature of the association between occupational physical activity (OPA) and leisure-time physical activity (LTPA). METHODS: 5448 adult men and women who participated in NHANES 1999-2000 were included in analyses. Linear regression was used to examine the bivariate and multivariable associations of OPA and education with LTPA. Generalized logit models were used to examine the association of education with OPA. RESULTS: We found no association between education and LTPA. OPA was significantly positively associated with LTPA (P < .001). The association between OPA and LTPA was not strongest among those with low education and held only for men in gender-stratified analysis. Education was inversely associated with OPA (P < .001) in multivariable analysis. CONCLUSIONS: Our findings lend preliminary support to the hypothesis that OPA is an important determinant of LTPA, particularly in men. This provides additional support to calls for assessment of OPA, particularly among individuals of low social class. PMID- 18382033 TI - Association of age and education with different types of leisure-time physical activity among 4437 Finnish adults. AB - BACKGROUND: The aim of this study was to investigate the associations of age and education with types of leisure-time physical activity in a population sample of Finnish adults. METHODS: The sample, part of the National FINRISK Study, comprised 1940 men and 2497 women age 25 to 64 years. Self-reported questionnaires were used to collect data on education and leisure-time physical activity. Overall leisure-time physical activity was further divided into conditioning and daily physical activity. RESULTS: In men and women, age had an inverse association with conditioning physical activity but not with daily and overall leisure activity. Strong direct associations were found between education and conditioning and overall leisure activity. CONCLUSIONS: All age groups report fairly similar levels of overall and daily levels of leisure-time physical activity, but the levels differ across educational groups. In health promotion, more emphasis should be targeted to the population groups with lower education. PMID- 18382035 TI - Expectancy beliefs and perceived values of Chinese college students in physical education and physical activity. AB - BACKGROUND: The expectancy-value theory postulates that motivation relies on individuals' beliefs of success, perceived Attainment, Intrinsic Interest, and Utility values and Cost. This study examined Chinese college students' expectancy value motivation in relation to physical education and self-initiated physical activity. METHODS: A random sample of 368 Chinese university students responded to questionnaires on perceived expectancy beliefs, perceived values, and cost in terms of their experiences in mandatory physical education programs and in self initiated after-school physical activity. They reported their choice decisions for continuing physical education. Data were analyzed using descriptive statistics, X2, logistic and linear regressions. RESULTS: The physical education curriculum was perceived as a major cost to motivation. Motivated by the Intrinsic Interest and Utility value, most students chose to continue to take physical education. Self-initiated after-school physical activity was motivated by the Attainment value only. No association was found between self-initiated physical activity and Liking or Disliking of physical education. CONCLUSION: Motivation for physical education and for self-initiated physical activity derived from different perceived values. The Attainment value motivates the students for self-initiated physical activity, whereas Intrinsic Interest and Utility values motivate them to choose to continue physical education. PMID- 18382034 TI - Do recreational resources contribute to physical activity? AB - BACKGROUND: Previous research suggests that access to recreational resources might influence physical activity. Little research, however, has looked at both access to and the characteristics of recreational resources and physical activity. METHODS: Access to recreational resources was assessed by counting the number of recreational resources in the geographic area. Resource characteristics were assessed through systematic observation (audits) or telephone interview of each resource. Access and characteristics in 2 counties in the St Louis, MO, metropolitan area with different prevalence rates of physical activity were compared using the critical-ratio (Z) test with P value for the difference between 2 independent proportions, given that the count and sample size were used to assess differences in access to equipment and presence of physical disorder. Financial accessibility was assessed for each facility. RESULTS: Data indicated significant differences in access and characteristics between the 2 areas that mimic differences in levels of physical activity. CONCLUSION: Our findings suggest that both access to and characteristics of recreational resources can contribute to differential rates of physical activity. PMID- 18382036 TI - Predictors of physical activity in the transition after high school among young women. AB - PURPOSE: The purpose of this study was to identify factors associated with physical activity (PA) in women during the first year following high school. METHODS: Females from 22 high schools (n = 915) completed the 3-Day Physical Activity Recall in 12th grade and reported if they were sports participants. After graduation, 305 women (18.9 +/- 0.6 years) completed the International Physical Activity Questionnaire. They reported time spent per day in moderate-to vigorous PA (MVPA) and vigorous PA (VPA) for the previous week. Multiple logistic regression was used to predict postgraduate PA. RESULTS: The odds of being in the high-active group were greater in women who were sports participants (OR = 1.93) in 12th grade. The odds of being in the high-active group were greater among white women (OR = 2.09) and greater among currently employed women compared with unemployed women (OR = 5.57). MVPA had borderline significance in the regression model. CONCLUSION: Sports participation and being currently employed predicted physical activity at postgraduation. PMID- 18382037 TI - Differences in leisure-time activities according to level of physical activity in adolescents. AB - BACKGROUND: The main goals of this study were: (1) to examine the relationship between physical activity (PA) involvement and other leisure activities in a sample of Portuguese youth and (2) to analyze gender differences in PA and leisure-time activity structure. METHODS: The sample comprised 1123 adolescents that were classified according to PA levels as active (n = 589) and nonactive (n = 534). A questionnaire assessing leisure-time activities was used. RESULTS: Girls were significantly more engaged in social leisure, dutiful, and individual artistic activities during leisure time, whereas boys were more involved in sports and computer and TV viewing activities. Significant associations between PA and social leisure were found in girls (r = .18, P < or = .001) and boys (r = .13, P < or = .01) after adjustment for age. The same was found between level of PA and sports engagement during leisure (girls: r = .56, P < or = .001; boys: r = .51, P < or = .001). In girls (r = .10, P < or = .05), but not in boys, a statistically significant association was found between PA and individual artistic activities. CONCLUSIONS: This study has certain implications for health related PA promotion efforts. Our data give additional reinforcement to the importance of organized and nonorganized sports/PA during leisure time for overall levels of PA in adolescents. PMID- 18382038 TI - Physical activity, stress, and metabolic risk score in 8- to 18-year-old boys. AB - BACKGROUND: We examined whether physical activity modifies the relationship between stress and the metabolic risk score in 8- to 18-year-old males (n = 37). METHODS: Physical activity (PA) and television (TV)/videogame (VG) use were assessed via accelerometer and questionnaire, respectively. Stress was determined from self-report measures. A metabolic risk score (MRS) was created by summing age-standardized residuals for waist circumference, mean arterial pressure, glycosylated hemoglobin, and high-density lipoprotein cholesterol. RESULTS: Correlations between PA and MRS were low (r < -.13), and TV and VG were moderately associated with MRS (r = .39 and .43, respectively). Correlations between stress-related variables and MRS ranged from r = .19 to .64. After partitioning by PA, significant correlations were observed in the low PA group between school- and sports-related self-esteem and anxiety with the MRS. CONCLUSIONS: The results provide suggestive evidence that PA might modify the relationship between stress and MRS in male adolescents. PMID- 18382039 TI - Inflated perceptions of physical activity after stroke: pairing self-report with physiologic measures. AB - BACKGROUND: Self-report measures of physical activity have well-known drawbacks, and physiologic measures alone do not account for behavioral variables important in the perception and performance of physical activity. Therefore, we considered multiple measures to quantify physical activity in community-dwelling men and women with chronic stroke. METHODS: This analysis included data from a volunteer sample of 87 individuals at least 6 months poststroke. Physical activity was measured using self-report questionnaires, step activity monitors, self-efficacy expectations related to exercise, and VO(2)peak from treadmill testing, and a model of physical activity was tested. RESULTS: Most of the variance in objective physical activity was explained by VO(2)peak, and most of the variance in subjective physical activity was explained by self-efficacy expectations. There were significant discrepancies between subjective and objective findings. CONCLUSION: This study helps to understand the perspective of stroke survivors with regard to physical activity. PMID- 18382040 TI - Where do the children play? The influence of playground equipment on physical activity of children in free play. AB - BACKGROUND: Promotion of physical activity in children depends on an understanding of how children use play equipment. METHODS: We conducted observations over 2 years of children in 2nd through 8th grades in a schoolyard with 5 distinct play areas with different amounts of play equipment. RESULTS: Children were more likely to play in areas with more installed play equipment, with densities of children in equipped areas 3.3 to 12.6 times higher than in an open grassy field. There were no significant differences by play area in the percent of children who were physically active at all, but children were more likely to be very active in areas with basketball goals and an installed play structure than in an open field. CONCLUSIONS: Playground equipment appeared to have a strong influence on where children played and a moderate influence on levels of activity. To maximize physical activity in children, playgrounds should be designed with ample and diverse play equipment. PMID- 18382041 TI - Reliability and clinical validity of a technique to assess lifetime illicit injection drug use. AB - A lifetime injection drug history is necessary to examine the impact of injection drug use on a physical health problem but it may cover time periods for which information and/or data reported may not be reliable. A test-retest study design was used to examine a technique of questioning persons about lifetime illicit injection drug use history (the Lifetime Injection History Questionnaire), including its reliability and relation to chronic venous disorders as an assessment of validity. Study participants included 104 persons (60 men, 44 women, M age = 49.3 years) provided services at a methadone maintenance treatment center located in a large industrial city in the Midwest. Kappa values for "ever injected" drugs ranged from 1.00 for heroin to .50 for nonprescription methadone (median = .75). High interclass correlations were found for youngest and oldest ages of injecting, years not injecting, and total injecting years (.90 to .98). Interclass correlation values for years injecting in the upper body and lower body were .79 and .70, respectively. Interrater reliability for the clinical portion of the venous disease assessment tool (the Clinical-Etiology-Anatomy Pathophysiology - CEAP - classification) was high: .97, right leg; .94, left leg. Controlling for age, gender, comorbidities, and body mass index, a classification of injection drug use based on the Lifetime Injection History scales accounted for 32% of the variance in the clinical CEAP scores. This is the first study to examine years of injection drug use that takes periods not injecting drugs into consideration. Focused substance abuse questioning (eg, drug, route, years of use) may help clinicians evaluate health problems related to drug use. PMID- 18382042 TI - Results of nine international pressure ulcer prevalence surveys: 1989 to 2005. AB - Pressure ulcers continue to be a significant problem for patients and healthcare facilities. Since 1989, results from the International Pressure Ulcer Prevalence surveys--observational, cross-sectional cohort studies--conducted by Hill-Rom, Batesville, Ind, have been used to document aggregate prevalence rates and provide acute care, long-term acute care, and long-term care facilities with internal and external benchmarks of process improvement. During each of the nine surveys conducted between 1989 and 2005, clinical teams in participating facilities predominantly in the US (some facilities in Canada, Saudi Arabia, and Australia participated after 2003) assessed admitted patients on assigned study dates. For this study, trends using all records (n = 447,930; average, 49,770 per year) were reviewed. The majority of facilities in each survey year were in the US (99% overall). Overall and nosocomial pressure ulcer prevalence rates ranged from 9.2% and 5.6% in 1989 to 15.5% and 10% in 2003 and 2004, respectively. The highest prevalence was documented in long-term acute care (27.3% overall, 8.5% nosocomial). Most commonly, ulcers were located at the sacrum (28%), heels (23.6%), and buttocks (17.2%). Ulcers were more commonly assessed as Stage I and Stage II (>70%). However, in patients with dark skin tone (2004 and 2005 data, n = 162,296), 13% of identified ulcers were Stage I compared to 32% in patients with medium and 38% in patients with light skin tone. Using the most complete data sets (2003, 2004, and 2005), more severe pressure ulcer prevalence (Stage III+) was not found to be age-related. Approximately 48% of all patients who had pressure ulcers and 48% of patients with nosocomial pressure ulcers were assessed at mild or no risk (Braden scale score >14). Prevalence within the Braden Score risk categories aligned with risk for developing pressure ulcers. Despite increased attention to the pressure ulcer problem, prevalence rates from the last five survey years are relatively unchanged. PMID- 18382043 TI - A descriptive study of pressure ulcer pain. AB - Despite increased emphasis on addressing patient pain, knowledge and information about pressure ulcer pain remains limited. To describe the quantitative and qualitative characteristics of pain related to pressure ulcers, a study was conducted among 47 hospitalized patients with pressure ulcers using the McGill Pain Questionnaire and Revised Faces Rating Scale. Volunteer participants, admitted for neurological disorders, ranged in age from 38 to 72 years (mean age 60.1 years +/- 8.23; 29 men, 18 women), six with Stage II, 32 with Stage III, and nine with Stage IV pressure ulcers. All but three (44, 94.6%) reported pressure ulcer pain; of those, 28 (59.5%) reported constant pressure ulcer pain and 41 (87.2%) did not specify when pain occurred. Dressing change, movement of the afflicted area, and pain at rest were reported by 32 (68.1%), nine (19.1%), and three (6.4%) patients, respectively. "Hot-burning" was the sensory descriptor most frequently used to describe the pain in 52%, 56%, and 67% of Stage II, Stage III, and Stage IV patients, respectively. Three out of six patients with Stage II ulcers rated their pain "discomforting," 32 of 32 with Stage III ulcers rated pain as "distressing," and nine out of nine with a Stage IV ulcer rated their pain as "horrible". Based on the Revised Faces Rating Scale, mean pain intensity was 6.04 +/- 2.78 (range 1 to 10), corresponding to moderate pain. For overall pain intensity, Patient Pain Index and Faces Rating Scale-Revised scores were highly correlated (r = 0.90, P <0.001). Pressure ulcers are painful, most patients report pain as "constant", and pain assessment should be included in all patient care plans. PMID- 18382044 TI - Decreasing the incidence of heel pressure ulcers in long-term care by increasing awareness: results of a 1-year program. AB - Heel pressure ulcers are a major problem in healthcare today. They involve extended clinician time, patient discomfort, and increased healthcare costs. In an attempt to decrease the incidence of heel pressure ulcers in one long-term care facility, a 1-year program was implemented that involved residents of one long-term care unit. In addition to staff education and awareness interventions, residents' heels were assessed daily and heel pressure-relieving measures were implemented. Kites were used to identify pressure ulcer stage, photo posters helped staff visualize the stages of heel pressure ulcers, and pencils marked with "Float Heels" were used to remind staff of the importance of prevention. Ulcer incidence rates were calculated every month. All residents (n = 40, mean age 67 years) on the unit at the beginning of the program were followed for as long as they were in the facility. At the start of the program, 50% of residents were at risk for developing ulcers and 22.5% had a heel ulcer. While the at-risk profile of residents remained relatively unchanged, no new ulcers were documented during nine of the subsequent 13 months, with incidence rates in the other 4 months ranging from 2.6% to 9.1%. Program costs were minimal and the results seem to confirm previously published studies about the positive effects of a comprehensive approach to the pressure ulcer problem. PMID- 18382045 TI - Topical metronidazole for the treatment of wound odor: a review of the literature. AB - Unpleasant odor is a frequent and distressing concern of persons who have wounds, as well as their family members and caregivers. In some instances, standard nursing interventions such as increasing the frequency of dressing changes will not be sufficient to address wound malodor. A review of the literature to summarize research findings about topical metronidazole used to decrease wound odor was conducted. Fifteen (15) studies were identified: seven case reports/series; six descriptive longitudinal studies; and two controlled clinical trials. Metronidazole was used as a 1% solution or, more frequently, as a 0.75% or 0.80% gel. Two reported adverse effects were skin irritation and a burning sensation. Generally, topical metronidazole was reported to result in a reduction or eradication of wound odor, decrease in wound drainage, improvement in wound appearance, decrease in surrounding cellulitis, halting of tissue necrosis, and decrease in pain. Although nurses report success with sprinkling crushed metronidazole pills on wounds, no published reports of this method of topical application are available. Topical metronidazole may be considered as an option for the management of malodorous wounds and additional randomized controlled trials with adequate sample sizes and comparison to products noted to decrease odor are warranted. While topical metronidazole for the treatment of malodorous wounds presently is off-label, perhaps such studies will result in the addition of malodorous wounds to the list of its indications. PMID- 18382046 TI - Prevalence of silver resistance in bacteria isolated from diabetic foot ulcers and efficacy of silver-containing wound dressings. AB - Silver dressings are used to manage wounds at risk of infection or locally infected. This in vitro study was conducted to assess the prevalence of silver resistance genes in 112 bacterial isolates obtained from the diabetic foot ulcers of patients attending the Diabetic Foot Clinic at Tameside General Hospital, UK. Using polymerase chain reaction to screen for three silver-resistance transcriptional units--silE, silS and silP--two silver-resistant bacteria were identified; both are strains of Enterobacter cloacae, an organism rarely implicated as a primary pathogen in chronic wounds. No recognized wound pathogens (Staphylococcus aureus-24 isolates and Pseudomonas aeruginosa-nine isolates) were found to contain silver-resistant genes. Analysis of the efficacy of silver containing dressings on the silver-resistant strains of Enterobacter cloacae using confocal laser microscopy showed that, despite evidence of genetic resistance to silver, all strains were killed following a maximum of 48 hours of exposure to the dressings. Results suggest that presence of silver resistance genes is rare and that genetic resistance does not necessarily translate to phenotypic resistance to silver. While silver resistance in wound care should be monitored, the threat of widespread resistance is low and silver-containing dressings remain an extremely important tool in managing wound infection. PMID- 18382047 TI - A clinically relevant wound assessment method to monitor healing progression. AB - One of the most important principles of wound management is periodic assessment and documentation of wound healing. Documentation of healing progress over time allows providers to assess the effectiveness of care to maximize healing. Several methods to determine wound healing progress currently exist and include dimensional, visual, and physiological assessments. However, because existing tools often require correlation of subjective assessments, are time-consuming, and may not consider that wound healing occurs from the "bottom up," a more objective and quicker approach to monitor healing progression was pursued. The purpose of this case study is to describe a once pen-and-paper tool that has now been computerized (the Barber Measuring Tool) that builds a graphical representation of a patient's individual wound healing progress to facilitate clinical decisions regarding the patient's plan of care. The tool, which is currently used for all wound patients in the author's facility, calculates wound volume using a simple formula and tracks this measurement as a percent of baseline over time in the patient's chart. Although formal research to establish validity and reliability of this tool has yet to be conducted, the tool has been used with more than 400 patients and has provided an accurate representation of healing progress. Studies to support proliferating use of this tool are warranted. PMID- 18382048 TI - Short-stretch or four-layer compression bandages: an overview of the literature. AB - Venous ulcers are a common, costly occurrence. Treatment typically includes the use of short-stretch and four-layer bandages--both with demonstrated ability to increase venous return and healing rates. Which is superior is unclear. To determine whether short-stretch bandages or four-layer compression systems provide shorter healing times and fewer adverse events when used in adults with venous ulcers, a search of English-language literature was conducted using the search terms short-stretch bandages and wound care, compression bandages and wound care, compression and venous ulcers, four-layer compression and venous ulcers, and multilayer compression and venous ulcers. Data from 25 studies published between 1997 and 2006 were examined. Short-stretch and four-layer compression bandages were found to be equally as effective in managing the edema that compromises venous ulcer healing. Time to healing was found to be shorter using four-layer or one- to two-layer compression when compared to paste compression in 24 studies. Ankle brachial index is an important factor to consider in determining compression appropriateness. Generally, use of any compression system is better than no compression at all. PMID- 18382049 TI - Multichannel surface recordings on the visual cortex: implications for a neuroprosthesis. AB - Using a multi-channel platinum surface electrode array, recordings from cat primary visual cortex were obtained in response to visual stimuli, and electrical stimuli delivered using the elements of the array itself. Neural responses to electrical stimuli were consistent, regardless of stimulus polarity or leading phase (biphasic), although thresholds were lower for monophasic than biphasic pulses. Both visual and electrical stimuli reliably evoked responses with characteristic components, which interacted with each other in a nonlinear summation showing first facilitation then suppression during the window of interaction. The chronaxie for eliciting threshold cortical responses was about 100 mus, and the charge density with a pulse width of 50-100 mus was around 55 muC cm(-2). These data form the basis of understanding the types of cortical responses to stimuli delivered by devices suitable for chronic implantation. PMID- 18382050 TI - [Construction and screening of genomic library from Raji cells]. AB - OBJECTIVE: To construct the genomic library of Raji cells and screen it by EBV DNA probe. METHODS: High molecular weight genomic DNA of Raji cells was digested by restriction enzyme BamHI. DNA fragments ranging from 9 to 23 kb were recovered by agarose gel electrophoresis, which were ligated with Lambda DASH II vector BamHI arms pre-treated with calf intestine alkaline phosphatase (CIAP). Ligated DNA was packed in vitro using Gigapack III gold packaging extract. The library was plated on XL1-blue MRA (P2) host strain.Titering and screening of the Raji genomic library were performed. RESULTS: The primary titer of the Raji genomic library was 1.8 x 10(5) pfu/mL, while that of the amplified library was 2.8 x 10(8) pfu/mL. Plaques (1 x 10(5)) were screened with (32)P-labeled EBV DNA probe(EBV genome 5-3271), 4 positive clones were obtained, and 1 of the 4 positive clones was picked out randomly for the second round of plaque screening. All the phage plaques were positive. DNA of the positive clone was extracted and was digested with BamHI. The length of the inserted fragment was 8.5 kb. Sequencing and BLAST analysis revealed that the inserted fragments consisted of the BamHI-W fragment at one end and clone RP11-665A22 on chromosome 15 at the other end. CONCLUSION: The successfully established genomic library of Raji cells will provide a basis for cloning the sequences of the EBV junction sites and interpreting the mechanism of oncogenesis of EBV integration. PMID- 18382051 TI - [Bone marrow endothelial cell-conditional medium promotes the generation of hematopoietic colony-forming cells from murine embryonic stem cells]. AB - OBJECTIVE: To observe the inductive efficiency of deriving hematopoietic colony forming cells from murine embryonic stem (mES) cells co-cultured with bone marrow stromal cell-conditional medium (mBMEC-CM). METHODS: After the day-4 embryoid bodies (4 dEBs) were derived from embryonic stem cell-D3 (ES-D3) cells, the cells of 4 dEBs were induced into hematopoietic colony-forming cells by co-culturing with mBMEC-CM. The numbers of 4 dEB-derived hematopoietic colonies (high proliferation potential-colony formation cells and burst forming unit-erythroid, HPP-CFC and BFU-E) were detected to explore the relation between the implanted 4 dEB-derived cell numbers and the colony numbers of BFU-E and HPP-CFC. The inducing effect of mBMEC-CM was observed according to the doses and days of induction. RESULTS: The number of 4 dEB-derived cells within 1 x 10(7)-4 x 10(7)/L was positively related to the colony numbers of HPP-CFC and BFU-E (HPP CFC, r=0.916,P< 0.05; BFU-E, r=0.927, P<0.05). The inducing doses of mBMEC-CM within 0-20% were positively related to the colony numbers of HPP-CFC and BFU-E (HPP-CFC, r=0.909, P<0.05; BFU-E, r=0.927, P<0.01). The colony numbers of HPP-CFC and BFU-E derived from the 4 dEB-derived cells were the highest after 3 days of induction, followed by those of 6 days and 9 days. CONCLUSION: Bone marrow endothelial cell-conditional medium can promote the generation of HPP-CFC and BFU E from murine embryonic stem cells. PMID- 18382052 TI - [Effect of IGF-1 on NO and PGE2 in rabbit articular chondrocytes induced by IL 1]. AB - OBJECTIVE: To explore the effect of insulin-like growth factor (IGF-1) on the concentration of NO and PGE(2) in the supernatant of rabbit articular chondrocytes induced by IL-1, and to explore the mechanism of IGF-1 in the development of osteoarthritis (OA). METHODS: The samples were divided into 7 groups: IL-1beta 10 microg/L group, IL-1beta 10 microg/L+IGF-1 1 microg/L group, IL-1beta 10 microg/L+IGF-1 10 microg/L group, IL-1beta 10 microg/L+IGF-1 50 microg/L group, IL-1beta 10 microg/L+IGF-1 100 microg/L group, IGF-1 50 microg/L group, and a blank control group. The chondrocytes from the articular cartilage of 2 month old rabbits were cultivated and identified, and then co-cultured in the second filial generation chondrocytes on plates with or without recombinant human IGF-1 or IL-1. The concentration of NO was detected by nitrate reductase kit, and that of PGE(2) by enzyme-linked immunosorbent assay (ELISA). The results were analyzed by statistical method. RESULTS: The average value of NO and PGE(2) was (89.971+/-10.224) micromol/L and (22.028+/-8.731) micromol/L in the IL-1beta 10 microg/L group, and (12.404+/-8.809) micromol/L and (1.900+/-0.227) ng/L in the blank control group. The concentration of NO and PGE(2) in IL-1beta 10 microg/L group was significantly higher than that in the blank control group (P<0.05). At the same concentration of 10 microg/L, IGF-1 could dose-dependently decrease the increase of NO and PGE(2) concentration induced by IL-1beta in the chondrocytes supernatant in vitro, and the optimum concentration of IGF-1 was 50 microg/L. CONCLUSION: IL-1 can significantly increase the concentration of NO and PGE(2), and IGF-1 can dose-dependently decrease the concentration of NO and PGE(2) in the chondrocytes supernatant in vitro. The optimum concentration of IGF 1 was 50 microg/L. PMID- 18382053 TI - [Brief self-administered risk questionnaire for common nutrition-related diseases in middle school students]. AB - OBJECTIVE: To develop a self-administered risk questionnaire for common nutrition related diseases in middle school students. METHODS: Two phases were conducted to develop the questionnaire: scale development and validation. Phase 1 included 7 steps: (1) determining the objective, theoretical framework, principles and format for indicator generation; (2) setting up the preliminary indicator pool; (3) selecting indicators and forming pilot questionnaire through focus groups; (4) testing the pilot questionnaire; (5) further correcting the questionnaire using expert consultation; (6) choosing indicators again using good-poor analysis; and (7) shaping the final questionnaire. Phase 2 consisted of: (1) using the Pearson correlation coefficient to assess test-retest reliability; (2) using the Cronbach's alpha coefficient to assess the internal consistency reliability; (3) using the feedback from field investigation to assess face validity; and (4) using explanatory factor analysis to assess construct validity. Students from 96 classes were selected at random in Hunan Province as the field test samples using stratified sampling and cluster sampling. And the students from 4 out of the 96 classes were chosen again to serve as the test-retest samples. We used Epidata 3.0 to build the database and SPSS 11.0 to analyze the data. RESULTS: A brief self-administered risk questionnaire for common nutrition related diseases in middle school students with 12 items being formed after Phase 1. Good-poor analysis showed results from t tests for each item were statistically significant (P<0.05). The Pearson correlation coefficient was 0.76 (P<0.05) and the Cronbach's alpha coefficient was 0.56. The questionnaire was accepted by the students participating in the field test. Four common factors were extracted using explanatory factor analysis, accounting for 50.18% of the total variation. CONCLUSION: The brief self-administered risk questionnaire for common nutrition-related diseases in middle school students is reliable and valid. PMID- 18382054 TI - [Construction and expression of recombinant adeno-associated virus vector containing HSV1-TK gene]. AB - OBJECTIVE: To construct the recombinant adeno-associated virus(rAAV) vector plasmid pSNAV2.0-TK containing HSV1-TK gene, to produce recombinant adeno associated virus rAAV2/HSV1-TK, and to detect the integration and expression of HSV1-TK gene in lens epithelial cells transfected by rAAV2/HSV1-TK, and to provide foundation for gene therapy of posterior capsular opacification. METHODS: The recombinant vector plasmid constructed by gene recombinant technology was analyzed by PCR and restriction enzyme digestion. The cell strain BHK-21/TK was screened by G418 after the plasmid was transfected into BHK-21 cells,with the helper virus HSV1-rc/UL2 to produce the recombinant virus rAAV2/HSV1-TK. The purity of rAAV2/HSV1-TK was detected by SDS-PAGE and HPLC, and the titre of rAAV2/HSV1-TK was observed by dot blot hybridization. The HSV1-TK gene in lens epithelial cells transfected by rAAV2/HSV-TK was investigated by PCR and RT-PCR. RESULTS: The recombinant plasmid proved successful by PCR and restriction enzyme digestion. The recombinant virus rAAV2/HSV1-TK was produced successfully and its titre was 1 x 10(12) v.g./mL by dot blot hybridization. The HSV1-TK gene was integrated and expressed in lens epithelial cells. CONCLUSION: The recombinant adeno-associated virus vector plasmid containing HSV1-TK gene is successfully constructed, and high titre recombinant adeno-associated virus (rAAV2/HSV1-TK) is obtained. The HSV1-TK gene in lens epithelial cells is expressed after being transfected by rAAV2/HSV1-TK. PMID- 18382055 TI - [Molecular mechanism of reversing multi-drug resistance of K562/AO2 by puerarin]. AB - OBJECTIVE: To determine the molecular mechanism of reversing multi-drug resistance of K562/AO2 by puerarin. METHODS: Effects of ADR and puerarin on NF kappaB activity of K562,K562/AO2 were tested by immunofluorescence. The expression of survivin of K562,K562/AO2 was examined by immunocytochemistry. The p-gp expression was detected by flow cytometry. RESULTS: The NF-kappaB activity of K562 was significantly higher than that of K562/AO2. The NF-kappaB activity of K562 treated by ADR was significantly higher than untreated. The NF-kappaB activity of K562 which was pretreated by puerarin and then treated by ADR was much lower than that treated by ADR alone. The NF-kappaB activity of K562/AO2 intervened by puerarin was lower than that unintervened by puerarin.The p-gp and survivin expression of K562/AO2 was significantly higher than K562. The p-gp and survivin expression of K562 treated by ADR was higher than that untreated by ADR. But the p-gp and survivin expression of K562 which was pretreated by puerarin and then treated by ADR was much lower than that not pretreated by puerarin.The p-gp and survivin expression of K562/AO2 intervened by puerarin was lower than that unintervened by puerarin. The expression was negatively correlated to the duration of intervention. The inhibition effect demonstrated time dependence. CONCLUSION: The activation of NF-kappaB can increase the expression of p-gp and survivin, which may be part of the molecular mechanism of multi-drug resistance of K562. Puerarin can prevent and stop the multi-drug resistance in K562 and reverse the multi-drug resistance of K562/AO2 to ADR by inhibiting the activity of NF-kappaB and the expression of p-gp and survivin. PMID- 18382056 TI - [Expression of Ki-67, Bcl-2,Bax and caspase-3 in benign prostatic hyperplasia combined with prostatitis and their significances]. AB - OBJECTIVE: To detect the expression of Ki-67, Bcl-2, Bax and caspase-3 in simple benign prostatic hyperplasia (BPH) and BPH combined with prostatitis,and to evaluate the effect of inflammation on the development and progression of BPH. METHODS: All specimens were obtained from patients undergoing surgical resection of the prostate. The paraffin section of the specimens was stained with hemotoxyline and eosin, and observed under light microscope to examine the inflammation hispathological changes. Sixteen patients with simple BPH (Group A) and 42 patients with BPH combined with prostatitis (Group B) were included. Immunohistochemical analysis and Western blot were used to examine the expression of Ki-67, Bcl-2, Bax and caspase-3. RESULTS: The expression of Ki-67 and Bcl-2 was significantly higher in Group B than that in Group A (P<0.05), and caspase-3 expression was significantly lower (P<0.05). There was no difference in Bax expression between the 2 groups (P>0.05). CONCLUSION: Prostatitis can up-regulate Ki-67, Bcl-2 expression, and down-regulate the expression of caspase-3 in BPH. Prostatitis appeared to play an important role in the development of BPH by affecting the proliferation and apoptosis of the prostatic cells. PMID- 18382057 TI - [Pathological features and clinical manifestations in 313 children with nephropathy under 6]. AB - OBJECTIVE: To explore the relationship between pathological features and clinical manifestations in children with nephropathy under 6 years old. METHODS: Renal biopsy by rapid percutaneous puncturation was performed on 313 children under 6 who were all diagnosed clinically as kidney diseases of 14 different kinds. The specimens were divided into 3 parts for microscope, electron microscope and immuno fluorescence examination respectively and processed by HE, PAS, PASM, and Masson staining. Immunofluorescence was used to detect the deposition of IgG, IgM, IgA, C3, C4, C1q, and Fb in the renal tissues. Additional examinations were done to detect HBs-Ag, HBeAg and HBcAg deposition in some cases with positive serum HBs-Ag. Altogether 290 of the specimens (290/313, 92.65%) were examined by electron microscope. RESULTS: All the renal biopsy performances were successful. The clinical manifestations comprised of persistent haematuria (32.92%, 103/313), idiopathic nephritic syndrome (26.1%, 82/313), acute nephritic syndrome (20.14%, 63/313), Henoch Schonlein purpura nephritis (8.32%, 26/313), HBV-nephritis (4.79%, 15/313), and isolated proteinuria (2.56%, 8/313). The main pathological patterns of glomerular disease were identified as mesangial proliferation (51.75%, 162/313), IgM nephropathy (8.31%,26/313), minor and minimal change (7.99%, 25/313), IgA nephropathy (7.35%, 23/313), endocapillary proliferative glomerulonephritis (5.11%, 16/313), focus segmental glomerulosclerosis (4.47%, 14/313), thin basement membrane nephropathy (4.47%, 14/313), and membrane nephropathy (4.47%, 14/313). Alport syndrome, congenital nephrotic syndrome, and thin basement membrane nephropathy can be diagnosed by electron microscope, white IgA nephropathy, IgM nephropathy and C1q nephropathy by immunopathology. CONCLUSION: Similar clinical manifestations may differ in the pathology and the clinical features of one pathological diagnosis may vary greatly. Renal biopsy is of great help to the diagnosis, treatment and the prognosis evaluation for children with nephropathy under 6. Electron microscopes also play an important role in the diagnosis of nephropathy. PMID- 18382058 TI - [Detection of autoantibodies in the sera of nasopharyngeal carcinoma patients]. AB - OBJECTIVE: To investigate whether there are autoantibodies to nasopharyngeal carcinoma (NPC) in the sera of patients and to find new NPC biomarkers. METHODS: Cell plate of Epstein Barr virus negative NPC cell line CNE1 was prepared, and difference between 32 NPC patient sera and 54 normal sera was analyzed by ELISA. We extracted the total protein of CNE1, and analyzed whether there were specific proteins to react with NPC sera by Western blot. RESULTS: The average absorbance values of serum antibody in NPC patients (0.904+/-0.032) were significantly higher than those of normal serum antibody absorbance values (0.736+/-0.028) (P< 0.01). The analysis with Western blot showed there were positive bands,and some of these were unanimous bands, but the intensity increased, and some of these were new bands compared with the normal sera. These positive bands may be NPC tumor-associated antigens or NPC tumor-specific antigens. CONCLUSION: Autoantibodies that react with NPC exist in the sera of NPC patients, but they do not react with Epstein-Barr virus. It provides the basis to seek the tumor biomarkers in NPC sera. PMID- 18382059 TI - [Effect of different glucose concentrations on the expressions of insig-1 and insig-2 mRNA during the differentiation of 3T3-L1 cells]. AB - OBJECTIVE: To determine the effect of different concentrations of glucose on the differentiation of 3T3-L(1) and the expression of insig-1 and insig-2 mRNA, and to explore the effect of insulin-induced gene in the differentiation and formation of adipocytes and lipogenesis. METHODS: The 3T3-L(1) cells were induced to differentiate in high glucose concentration (25 mol/L G.S), low glucose concentration (5.5 mol/L G.S), and mannitol (19.5 mol/L Mannitol +5.5 mol/L G.S), respectively. The differentiation of 3T3-L(1) cells was examined by oil red "O" straining, and the expression of insig-1,insig-2 mRNA and AP2 mRNA was examined by RT-PCR and in situ hybridization. RESULTS: With the differentiation of 3T3 L(1) cells, the expression of insig-1 and insig-2 mRNA was gradually up regulated. The expression of insig-1 and insig-2 mRNA significantly increased while AP(2) mRNA decreased in the low glucose concentration inducing group and mannitol inducing group. In the high glucose concentration inducing group, the cell differentiation was poor (P<0.05). There was no difference between the low glucose concentration and the mannitol group in the differentiation of 3T3-L(1) cells, and in the expression of insig-1 and insig-2 and AP(2) mRNA. CONCLUSION: Different concentrations of glucose may influence the cell differentiation and the low glucose concentration promotes insig-1 and insig-2 gene expression, which may lead to the inhibition of the differentiation and lipogenesis of preadipocytes. PMID- 18382060 TI - [Therapeutic effect of priming induction regimen of CAG for newly diagnosed acute myeloid leukemia in elderly patients]. AB - OBJECTIVE: To evaluate the clinical efficacy and toxicity of priming induction regimen of CAG for newly diagnosed acute myeloid leukemia (AML) in elderly patients. METHODS: Seventy-five patients with newly diagnosed AML were divided into 2 groups: 34 were treated with priming induction regimen CAG and the other 41 were treated with 2 classic routine chemotherapy regimens including pirarubicin+cytarabine (TA) and homoharringtonine+cytarabine (HA). All patients had a 14 day interval between the 2 courses of chemotherapy. RESULTS: The complete remission rate after 2 courses of induction therapy in patients with the priming induction regimen CAG and the total efficacy rate was significantly higher than that of the routine chemotherapy patients(67.6% vs. 39%; 82.4% vs. 56.1%). Patients with unfavorable karyotypes had poor chemotherapy efficacy. The 3-year disease-free-survival (DFS) time was longer in patients with AML treated with priming induction regimen CAG than in patients treated with 2 classic routine chemotherapy regimens. Except for the muscular soreness, the hematological and non-hematological side effects in the CAG priming induction group were significantly fewer than those in the routine chemotherapy group. CONCLUSION: The priming induction regimen of CAG has a significantly higher complete remission rate and an efficacy rate, fewer side effects, milder chemotherapy intensity and is more sensitive to chemotherapeutic drugs than those of the routine chemotherapy. It can shorten the duration of agranulocytosis and decrease infectious complications and increase the sensitivity of leukemia blast cells to chemotherapeutic drugs. PMID- 18382061 TI - [Expression of aquaporin-4 protein in rats with acute radiation-induced cerebral edema]. AB - OBJECTIVE: To explore the relation between changes in expression of aquaporin 4(AQP4) protein and acute radiation-induced cerebral edema after Gamma knife radiation. METHODS: The experimental model was established in rats by radiating 2 mm right to median line in cerebral hemisphere with 50 Gy Gamma knife (the center located at the line of auditory canal). The changes of brain water content were measured by the wet and dry weight method. Immunohistochemistry was used to determine the change of expression of AQP4 protein at different periods after Gamma knife radiation. RESULTS: The brain water content and the expression of AQP4 in the target area and the peripheral zone obviously increased at 12 h after Gamma knife radiation, and achieved the peak after 2 d. It was still higher than normal after 14 d. The gray value of expression of aquaporin-4 was negatively related to brain water content (r=-0.9857, P<0.05). CONCLUSION: AQP4 has a close relationship with acute radiation-induced cerebral edema. PMID- 18382062 TI - [Retrospective study on clinical application of biovalves in 52 cases]. AB - OBJECTIVE: To summarize the clinical application of bioprosthetic valve replacement. METHODS: Fifty two patients, aged 13-73(52.4+/-14.0) years, underwent cardiac valve replacement with biovalves from June 2002 to June 2006 in our hospital. Fifty three tissue valve replacements combined with 8 mechanical valve replacements were performed. Other procedures were also carried out if appropriate, including bidirectional Glenn shunt in 1 patient with double outlet of right ventricle and pulmonary artery stenosis; interruption of accessory pathway in 1 patient with Wolff-Parkinson-White syndrome; coronary artery bypass graft in 5 with severe coronary artery disease; and atrial or ventricular repair in 4 with congenital septum defects. RESULTS: One patient died in hospital due to multiple organ failure. The hospital mortality rate was 1.9%. The mean follow-up was 25 months. Two patients lost follow-up and the follow-up rate was 96%. Forty one patients returned to NYHA class I, 9 to class II, and 1 to class III. Echocardiography showed the implanted bioprosthesis functioned well without stenosis or regurgitation. There was no structural deterioration with freedom from thromboembolism and anticoagulant hemorrhage in all patients. CONCLUSION: The efficacy of biovalves in heart valve replacement is satisfactory. PMID- 18382063 TI - [Indications and effect of the intra-aortic balloon pump in 38 patients during the cardiac surgery]. AB - OBJECTIVE: To review the indications and effect of the intra-aortic balloon pump (IABP) in 38 patients during the cardiac surgery. METHODS: From June 2004 to August 2007, 38 patients were performed IABP in the cardiac surgery, including 28 coronary artery bypass grafting (CABG) and 10 valve replacements. The intra operation prophylactic use of IABP was in 5 cases, the use of IABP for weaning from cardiopulmonary bypass (CPB) during the cardiac surgery was in 22 cases, and the rest use of IABP was after the operation in the intensive care unit. Low molecular weight heparin was used for IABP anticoagulation. RESULTS: Thirteen died out of the 38 IABP patients. The total IABP success rate was 66%.The IABP success rate in CABG was 82%, and 20% in the valve replacement. The patients with intra-operation prophylactic use of IABP all recovered, and 16 recovered out of the 22 patients with IABP uses for weaning from CPB, and 6 recovered out of the 11 patients with post-operation IABP application. No severe complications related to IABP occurred. CONCLUSION: IABP has better effect in CABG patients than in valve replacements. Effect of the prophylactic use of IABP is better than both the IABP use for weaning from CPB or the post-operation IABP application. For IABP anticoagulation, low molecular weight heparin is the better alternative than routine heparin, and the proper ACT is controlled at 150-170 s. PMID- 18382064 TI - [Three subanaesthetic dose ketamines mixed with butorphanol in the postoperative continuous intravenous analgesia]. AB - OBJECTIVE: To determine an optimal clinical dose of ketamine after comparing the efficacy and security of 3 low dose ketamines mixed with butorphanol in the postoperative continuous intravenous analgesia. METHODS: Eighty ASA (American Society of Anesthesiologists) I-II patients scheduled for elective gynecological surgery under general anesthesia were divided randomly into 4 groups (n=20): Group B received butorphanol 3 microg/(kg x h);Group BK1 received butorphanol 2 microg/(kg x h) mixed with ketamine 60 microg/(kg x h); Group BK2 received butorphanol 2 microg/(kg x h) mixed with ketamine 90 microg/(kg.h); and Group BK3 received butorphanol 2 microg/(kg x h) mixed with ketamine 120 microg/(kg x h). Continuous intravenous infusion pump was used when the patients had obvious pain (visual analgesia scale of five), and the bolus infusion (4 mL) was given before the operation, and continuous infusion at 2 mL/h. In the postoperative period, pain was assessed using visual analogue scale (VAS) at 2,6,12,24, and 48 h.At the same time, Ramsay scores and adverse effects were recorded. RESULTS: There was no significant difference in the adverse effects and the postoperative mean arterial pressure, heart rate, respiratory rate values, and pulse oxygen among the 4 groups. Postoperative VAS values in Group BK3 was the lowest, followed by Group BK2. There was no significant difference between Group BK1 and Group B. The incidence of somnolence in Group B was higher than that in Group BK1, BK2 and BK3(P<0.05). CONCLUSION: Ketamine in subanaesthetic dose added to butorphanol for postoperative continuous intravenous infusion has a better postoperative analgesic effect and sedation. It can effectively spare butorphanol consumption without increasing adverse effects. The optimal combined dose is 90-120 microg/(kg x h). PMID- 18382065 TI - [Pneumonectomy for 210 cases of lung cancer]. AB - OBJECTIVE: To summarize 210 cases of lung cancer that underwent pneumonectomy,and to study the therapy value of pneumonectomy for lung cancer. METHODS: Between January 1993 and December 2004,210 patients (169 males and 41 females) with lung cancer underwent pneumonectomy in our hospital with the mean age of 54.77 years. Out of the 210 cases 144 had left pneumonectomy, and 66 had right pneumonectomy. Among them, 26 experienced intrapericardial pneumonectomy,1 inferior vena cava reconstruction, and 1 carina reconstruction. All bronchial stumps were closed by manual suture,and systemic node dissection was performed routinely. RESULTS: No one died during the operation, but 59 patients (28.10%) experienced postoperative complications and 9 patients (4.28%) died within 30 days of operation. After exclusion of postoperative deaths, the 1-year, 3-year, and 5-year survival rates were 66.21%,36.13% and 24.26%, respectively. Only 1 out of the 12 patients with small cell carcinoma survived more than 3 years. CONCLUSION: Mortality and morbidity were high in pneumonectomy.If we carefully and properly select the patients, enhance postoperative monitoring and perioperative treatment, and combine with chemo-and/or radiotherapy,pneumonectomy will surely be effective for patients with lung cancer. PMID- 18382066 TI - [Release of gluteal muscle contracture by radiofrequency under arthroscopy]. AB - OBJECTIVE: To evaluate the clinical therapeutic effect of the release of gluteal muscle contracture by radiofrequency under arthroscopy. METHODS: From January 2004 to April 2005, 86 patients with gluteal muscle contracture were treated by radiofrequency release under arthroscopy.The patients were followed-up for 6-18 months(mean=12.6 months). RESULTS: Getting carriage, squatting down while keeping their knees contacting, crossing leg test, and impact on movement were served as evaluation criteria. Of the 86 patients, 79 had excellent results, 5 good, and the rest 2 were acceptable. There was no recurrence at the last follow-up, and all the patients were satisfied. CONCLUSION: For gluteal muscle contracture, radiofrequency release under arthroscopy has minimally invasion, good results and quick recovery. PMID- 18382067 TI - Neuroendocrine, metabolic, and immune functions during the acute phase response of inflammatory stress in monosodium L-glutamate-damaged, hyperadipose male rat. AB - In rats, neonatal treatment with monosodium L-glutamate (MSG) induces several metabolic and neuroendocrine abnormalities, which result in hyperadiposity. No data exist, however, regarding neuroendocrine, immune and metabolic responses to acute endotoxemia in the MSG-damaged rat. We studied the consequences of MSG treatment during the acute phase response of inflammatory stress. Neonatal male rats were treated with MSG or vehicle (controls, CTR) and studied at age 90 days. Pituitary, adrenal, adipo-insular axis, immune, metabolic and gonadal functions were explored before and up to 5 h after single sub-lethal i.p. injection of bacterial lipopolysaccharide (LPS; 150 microg/kg). Our results showed that, during the acute phase response of inflammatory stress in MSG rats: (1) the corticotrope-adrenal, leptin, insulin and triglyceride responses were higher than in CTR rats, (2) pro-inflammatory (TNFalpha) cytokine response was impaired and anti-inflammatory (IL-10) cytokine response was normal, and (3) changes in peripheral estradiol and testosterone levels after LPS varied as in CTR rats. These data indicate that metabolic and neroendocrine-immune functions are altered in MSG-damaged rats. Our study also suggests that the enhanced corticotrope corticoadrenal activity in MSG animals could be responsible, at least in part, for the immune and metabolic derangements characterizing hypothalamic obesity. PMID- 18382069 TI - Body composition, changing physiological functions and nutrient requirements of the elderly. AB - Age has an important impact on body composition. The decreases of lean body mass and total body water accompanied by an increase of body fat are the most relevant changes leading to a reduction of the basal metabolic rate. Due to this and the fact that elderly people are generally less physically active, energy needs are lower. However, the requirements for most micronutrients are not reduced. Indeed, impaired absorption of some vitamins and minerals like cobalamin and calcium can cause deficiencies. An adequate calcium supply is particularly important as with ongoing age, bone mass decreases as well, making elderly prone to osteoporosis. The reduced ability to synthesize vitamin D and the lower sunlight exposure of many elderly further aggravate this issue. There is evidence to consider the supply of some important dietary antioxidants as critical in some elderly, namely vitamin C and beta-carotene as well as zinc. In light of age-related decreases in the activities of antioxidant enzymes, a sufficient supply of dietary antioxidants is important, especially as oxidative damage is thought to contribute to the deteriorating processes associated with aging and promote cardiovascular disease, cognitive disorders, cancer and diabetes mellitus that occur more frequently in older people. The first three are also associated with an insufficiency of folate, another critical nutrient in the elderly. Therefore, dietary guidelines should strive to optimize the immune function and reduce disease risk of the elderly. PMID- 18382070 TI - Aging, disability and frailty. AB - Despite multiple and often overlapping definitions of disability and frailty, both are common clinical characteristics of aged individuals though not identical. The geriatric syndrome of frailty is described as status of global impairment of physiological reserves involving multiple organ systems. The clinical correlate of frailty manifests as increased vulnerability, impaired capability to withstand intrinsic and environmental stressors, and limited capacity to maintain physiological and psychosocial homeostasis. Geriatric frailty is found in 20-30% of the elderly population over 75 years and increases with advancing age. It was reported to be associated with long-term adverse health-related outcomes - increased risk of geriatric syndromes, dependency, disability, hospitalization, institutional placement, and mortality. The clinical phenotype of frailty manifests as multi-system pathologies characterized by low physical activity, global weakness with low muscle strength, fatigability/exhaustion, overall slowness particularly of gait, loss of weight among others. These above-mentioned clinical symptoms could be explained by (or related to) some 'preclinical' diagnoses such as sarcopenia, osteopenia, nonspecific balance disorders, nutritional problems, and overall deconditioning. More recent studies found the frailty clinical phenotype to be associated with pathologic laboratory markers (IL-6, CRP, 25-hydroxyvitamin D, IGF-1, D-dimers), which suggest possible pathogenesis involving hormonal dysregulation, immuno aging, pro-coagulation and pro-inflammatory status. In the article, current recommendations for future research strategies of frailty syndrome will be discussed. PMID- 18382071 TI - Intestinal cholesterol conversion in adults and elderly from four different European countries. AB - The growing healthy aging population in Europe is a heterogeneous group with varying nutritional needs, especially in association with problems like decreased sense of smell and taste, saliva production, chewing and swallowing problems besides all disease problems that put specific demands on the dietary intake and the role of the intestinal metabolism. To investigate this, one must clarify which functions are related to the host itself and which actions are related to the microflora. In short, the host's side of the ecosystem can be defined as milieu interieur (MI), the non-host side as milieu exterieur (ME) and MI plus ME together as milieu total (MT). We have investigated the role of the intestinal flora and its functions in adults and elderly from different European countries and this publication is concentrated on the microbial conversion of intestinal cholesterol to coprostanol, aiming to investigate the hepatic-intestinal flora co cross-talk. PMID- 18382072 TI - Changes in food preferences during aging. AB - Due to the increasing proportion of the elderly in the European Union knowledge of health and nutritional status has to be complemented by studies focused on food preferences and health behavior. A comprehensive literature review has been conducted. The state of the art documents a gap in understanding why differences exist between food knowledge, attitudes and practices. Inadequate attention has been given to cultural factors. Research into the reasons for nutrition behavior and food choice is of key importance for the future. PMID- 18382073 TI - Quality of life, food choice and meal patterns - field report of a practitioner. AB - Quality of life is defined as the result of combining personal resources, control of the environment, personal values, and actual living conditions. Balanced nutrition is an important condition for quality of life, health and well-being. During the course of life everyone develops his very individual biography of eating. This includes eating habits, food choice, and meal patterns. The process of aging is accompanied by hardly recognizable physiological, emotional, social, and environmental changes. Ignoring these changes can lead to malnutrition and nutrition-related problems and thus reduce health, diminish the quality of life, and overall well-being. Accordingly, it is necessary to synchronize the individual biography of eating, the physiological, emotional, social, and environmental changes to enable the aged to feel self-determined and self confident. This presentation will describe successful examples from local homes for the aged/nursing homes. Examples will show that possibilities of food choice answer the need of the residents to control their environment, that residents can be integrated in the planning and preparation of food and how this corresponds with their need to show their knowledge and experience, that meals in residential facilities can be re-arranged to let the residents experience joy and pleasure, and that nutritional concepts in nursing homes can be changed so that the residents experience themselves as subjects of nutrition. PMID- 18382074 TI - Sensory design of foods for the elderly. AB - BACKGROUND/AIMS: Elderly persons with dysphagia need food that requires little or no chewing, that is easy to swallow and has attractive sensory characteristics. The aim was to investigate how ingredients varied according to experimental designs influence the perceived sensory, chewing and swallowing characteristics of two types of texture-modified model foods. METHODS: Meat- and carrot-based, texture-modified model foods were produced. The following parameters were varied: particle size, fat content, starch and egg composition. The samples were studied using sensory analyses, focus group discussions and consumer studies. RESULTS: The design parameters mainly had an impact on sensory texture attributes. The experts in the focus groups selected three products of each type which were regarded as being optimal for older persons. All the products contained a high proportion of egg yolk and a low amount of starch. Older consumers considered all the selected products to be easy to chew and swallow. The differences between older persons in nursing homes compared to those living in their own homes could be linked to health. CONCLUSIONS: Optimization of factors influencing food quality through the use of experimental designs in combination with sensory and consumer studies is required in order to meet the needs and demands of older people. PMID- 18382075 TI - Food and drug interaction: consequences for the nutrition/health status. AB - Food-drug interactions are defined as alterations of pharmacokinetics or pharmacodynamics of a drug or nutritional element or a compromise in nutritional status as a result of the addition of a drug. Elderly patients are particularly at risk because more than 30% of all the prescription drugs are taken by this population. Failure to identify and properly manage drug-nutrient interactions can lead to serious consequences. For instance, drug-nutrient interactions can result in reduced absorption of certain oral antibiotics and may lead to suboptimal antibiotic concentrations at the site of infection. This predisposes the patient to treatment failure. Induction or inhibition of enzymes in the gut by nutrients may lead to a significant change in oral bioavailability of drugs or vice versa. For example, grapefruit juice is a selective intestinal CYP3A4 inhibitor. The overall exposure of some drugs can be increased by more than fivefold when taken with grapefruit juice and increase the risk of adverse effects. The use of certain drugs may affect GI tract function and may lead to a loss of bodily electrolytes and fluid. Limiting drug prescriptions to essential medications for as short a period as possible and periodic re-evaluations of the treatment chosen are essential to minimize adverse drug-nutrient interactions. PMID- 18382076 TI - Nutritional supplementation in community-dwelling elderly people. AB - There is a large evidence base for nutritional intervention in acutely ill and post-operative hospitalised patients, but the evidence base for nursing home (NH) residents is small. The prevalence of poor nutrition in NHs is high and baseline nutrition appears to be an important determinant of response to nutritional intervention. Residents with mininutritional assessment (MNA) scores above 23.5 tend to show less response than those with lower scores. This relates in part to failure to increase intake in the better nourished as well as to actual response to increased intake. At the low end of the MNA spectrum, the increasing prevalence of multiple pathologies tends to result in a reduced response, but randomised controlled studies in this group is probably not ethical. Most studies have tended to investigate the intermediate group with MNA scores of 17-23.5 or equivalent using other scales. Interventions have usually resulted in increased intake of calories and micronutrients. Other end points have variously shown responses including weight, immunological measures, infection rates, decubitus ulcers, falls and fracture rates. Many studies have been too small to demonstrate benefit and some are likely to have suffered from type l errors - showing benefit by chance. Poorly quantifiable variables likely to be of importance include the local environment and catering as well as pathophysiological variability. PMID- 18382077 TI - Thanks to body exercise, getting mobile and being less dependent. AB - The study was aimed at determining the influence of aerobic endurance exercise and the intake of a multivitamin beverage (MV) on institutionalized people in their 8th and 9th decades of life. The study comprised 4 groups: exercise + MV (n = 16 completer, age: 83 +/- 8 years), exercise (n = 8, age: 82 +/- 8 years), MV (n = 8, age: 83 +/- 5 years), control (n = 7, age: 83 +/- 5 years). The period of exercise, 'aqua training', lasted 17 weeks, two times a week for 25 min. During this period, 250 ml/day of a commercially available multivitamin drink was administered in two of the four groups. At the outset and after the intervention, status of lipophilic antioxidants, vitamin C, malondialdehyde (MDA), cholesterol (total, LDL, HDL), triglycerides and uric acid was assessed. The influence on physical activity was evaluated via spiroergometry. The initial status of nutritive antioxidants was very satisfactory for this population group, only plasma beta-carotene was borderline. Plasma levels of total-cholesterol (243 +/- 42 mg/dl), LDL-cholesterol (161 +/- 35 mg/dl) and uric acid (5.8 +/- 1.4 mg/dl) were slightly above the reference value. Endurance exercise neither increased MDA nor decreased plasma antioxidants or changed plasma lipids. Moreover, plasma levels of beta-carotene (all MV intervention) and cryptoxanthin (group: exercise + MV) increased after intervention (p < 0.05). Physical performance did not enhance, only in the group exercise + MV the respiratory ratio (p < 0.05) and lactate (p = 0.072) increased. PMID- 18382078 TI - Food frequency index as a measure of diet quality in non-frail older adults. AB - AIMS: The objective of the present work was to identify factors associated with diet quality in non-frail older adults measured with the Food Frequency Index (FFI). METHODS: In a cross-sectional survey, data from a qualitative 28-item food frequency questionnaire was used to calculate the FFI in a sample of 444 adults aged 55 years and older living in Vienna, Austria. RESULTS: A higher FFI, indicating a favorable diet quality, was obtained for women as compared to men, younger and better-educated subjects (p for all < 0.01). Dietary supplement users (p = 0.04) and subjects being physically active (p < 0.01) showed significantly higher FFI scores. Eating difficulties (p = 0.04) and gastrointestinal problems (p = 0.03) were found to be risk factors for poor diet quality as measured with the FFI, whereas the number of prescribed daily medications was not associated with diet quality (p for trend = 0.77). CONCLUSIONS: Differences in diet quality are evident in subgroups of non-frail older adults living in Vienna as defined by societal factors, such as education, lifestyle-related factors, such as being physically active and health-related factors, such as oral health. PMID- 18382079 TI - Quality of nutrition of elderly with different degrees of dependency: elderly living in private homes. AB - BACKGROUND/AIM: To be aware of the influencing factors of healthy aging, several cross-sectional surveys have been carried out among the elderly living in Austria to identify health and lifestyle factors as well as socioeconomic influences that contribute to the nutritional situation. METHODS: Information about the health situation was collected by questionnaires. Diet was assessed by both qualitative food frequency questionnaires and 24-hour recalls. Additionally, nutritional status was examined in blood and urine samples using static and functional parameters. RESULTS: The prevalence of overweight (21%, BMI >29) and obesity increased with increasing age up to 65 years; in people older than 65 years an inverse tendency could be observed; as from this age upwards a high prevalence of underweight was found (22%, BMI <24). The consumption of meat and meat products was too high compared to the recommendations. By contrast, the intake of high nutrient density foods was insufficient. On average, the fat intake was around 38% of total energy (%E); that of saturated fatty acids amounted to 20%E; the intake of carbohydrate and fiber was considered as being too low. The intake of folate, vitamin D, calcium, iodine, magnesium and vitamin C was assessed as inadequate. CONCLUSION: More attention has to be paid to certain micronutrients in order to optimize dietary intake and to sustain health. PMID- 18382080 TI - Assessment of the nutritional situation of elderly nursing home residents in Vienna. AB - BACKGROUND: In Austria, reliable information about the nutritional situation of institutionalized, frail elderly is lacking. METHODS: MNA (Mini Nutritional Assessment) and NuRAS (Nutritional Risk Assessment Scale), tools specifically developed to evaluate the nutritional status of the elderly, were used for the assessment of the general nutritional and health status of the residents. Individual intake of energy and nutrients was measured by using the weighed food intake method during 7 consecutive weekdays. RESULTS: According to MNA results (n = 245), 13.9% had a satisfactory nutritional status, 48.3% were assessed as 'at risk of malnutrition', and 37.8% as being 'malnourished'. There is evidence that the number of assessed nutritional risk factors has an impact on the nutritional status. Further, there is a link between the assessed nutritional status and the individual nutrient intake of the residents. CONCLUSION: There is evidence for nutritional problems in frail, institutionalized elderly nursing home residents. The MNA seems to be a reliable tool to identify individuals at risk for malnutrition. Strategies for effective nutritional intervention should be implemented in long-term care facilities for the elderly in order to prevent and treat malnutrition and to ensure high-quality care. PMID- 18382081 TI - Intervention with dietary fiber to treat constipation and reduce laxative use in residents of nursing homes. AB - BACKGROUND: Residents of nursing homes have lost their independence. Recent studies reported that nutritional problems arise in nursing homes. These problems are correlated with changed eating habits and geriatric constipation which is predominantly treated with laxatives. These interventions are not always without risk since frequent usage of laxatives may be accompanied by several side effects. Dietary fibers also affect stool weight and transit time. Therefore, oat bran effectiveness in reducing the need for bowel medication was examined. AIM: To develop diets with the addition of oat bran for reduction of laxatives and to improve the inhabitants' well-being in a long-term-care facility. METHODS: A controlled blind parallel intervention trial among 30 frail inhabitants of a geriatric ward aged 57-100 years with laxative use. An intervention and a control group were formed. 15 of them received oat bran for 12 weeks (fiber group) mixed up in the daily common diet of the ward and 15 served as control (control group). Laxative use, body weight and the observations concerning the eating habits of the elderly were documented. RESULTS: A cake with the required dietary fibers and the complaisant acceptance of the seniors was developed. Laxatives were successfully discontinued by 59% (p < 0.001) in the fiber group. Body weight remained constant in the fiber group and decreased in the control group (p < 0.005). CONCLUSION: Fiber supplementation in the form of a cake allows discontinuation of laxatives and increases the seniors' well-being in a nursing home. PMID- 18382082 TI - Nutritional situation of the elderly in the European Union: data of the European Nutrition and Health Report (2004). AB - BACKGROUND: The European Nutrition and Health Report contains data about the nutritional situation of elderly of 11 EU countries. METHODS: The assessment of the energy and nutrient intake was performed using different methods: 24-hour recall, (semiquantitative) FFQ, 4/7-day-weighed record, personal interview, HBS. RESULTS: Compared to adults of the same country, the energy intake of elderly was on average lower. In most EU countries, the mean proportion of protein intake was within the recommended range while the share of carbohydrates was considerable low. The mean fat intake was remarkably high, the dietary fatty acid pattern predominantly imbalanced and the intake of cholesterol exceeded 300 mg/day in most European elderly. The average intake of vitamin D and folic acid was clearly below 10 and 400 microg/day, respectively, in elderly of all countries. Sodium intake was quite high in several participating countries, whereas calcium intake remained below 800 mg/d in all investigated states. The supply of vitamin A/C, thiamine, niacin, cobalamin, and the elements: magnesium, iron, zinc, copper, and manganese could generally be regarded as sufficient. Only in some countries the intake of these nutrients had to be assessed as unsaturated. CONCLUSION: Data analysis showed that the proportions of macronutrient intake of European elderly are not balanced. Apart from vitamin D, calcium, folate, iodine, and sodium the mean intake of micronutrients could be regarded as acceptable. PMID- 18382083 TI - Nutritional situation of the elderly in Eastern/Baltic and Central/Western Europe - the AgeingNutrition project. AB - BACKGROUND/AIMS: In the AgeingNutrition project, funded by the European Commission, partners from 10 'Eastern/Baltic' (new) European (Bulgaria, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Slovak Republic, Slovenia, and Turkey) and 5 'Central/Western' (old) European countries (Austria, Belgium, Germany, Greece, and Spain) performed a comparative analysis of existing data on nutritional status, food intake, nutrient intake, dietary habits and lifestyle of the aging population in Europe. METHODS: Using predefined inclusion criteria, 36 studies (22 from Eastern/Baltic, 14 from Central/Western countries) were chosen for evaluation. The studies varied in general design, data assessment and parameters and methods applied (especially regarding assessment of nutritional situation, comprising anthropometry, blood lipid profile, dietary nutrient and food intake). Thus, data evaluation by recalculation of raw data had to be restricted to a few distinct, common parameters. RESULTS AND CONCLUSION: Energy intake is too high in both regions, especially in some of the 'younger old' and seems not to be adjusted to energy expenditure. Simultaneously, energy intake was considerably lower than recommended in some very old seniors in Eastern/Baltic countries. Interestingly, quantity and quality of available information is similar in 'new' and 'old' member states. Immense variations in methodology hampered performing reliable comparative evaluation. It is, thus, mandatory to standardize methods for assessment of nutrition and health status for future international multicenter studies. PMID- 18382084 TI - Outcomes of the Nutri-Senex project: a co-ordination action funded by the European Commission. AB - This project's primary aim is to contribute to the improvement of the quality of life of the elderly. The state of the art in age-related research and technology, to highlight the most promising areas for development, has been documented in an extensive literature report. A research database has been established to map current research activities within the European research area. Through several expert group meetings with representatives from universities, research institutions, organizations concerned with the care of the elderly, and industry, suitable technologies and policies that can improve the diet of the elderly have been identified. A legislation survey relating to aspects of nutrition of the elderly has been carried out. A set of guidelines and recommendations of best practice for use in the health and care industry have been developed. The project's results are disseminated through an enhanced website, technology transfer workshops and a staff training program. Good communication between the partners has been promoted and permanent research links have been forged. The main outputs will lead to better co-ordinated European research: less duplication of effort, more rapid commercialization, increased awareness of the problems faced by the elderly, improved guidelines and recommendations for health care professionals and policymakers. PMID- 18382085 TI - Cytapheresis in inflammatory bowel diseases: current evidence and perspectives. AB - Ulcerative colitis and Crohn's disease are inflammatory bowel diseases with a chronic relapsing course. Management of both conditions is far from being fully satisfactory. For this reason in the last decade a large number of biological therapies, targeting cytokines involved in intestinal inflammation, has been developed with various results in terms of efficacy, safety and costs. Activated granulocytes and monocytes represent the major sources of pro-inflammatory cytokines in the intestinal mucosa, playing a pivotal role in inducing and maintaining intestinal inflammation. Leukocytapheresis using an adsorptive carrier-based system (Adacolumn) or a removal filter column (Cellsorba) has been proposed as a feasible, safe and effective therapy for ulcerative colitis and Crohn's disease. The objective of this paper is to provide an overview on the current knowledge about mechanisms of action, available clinical data and the possible future perspectives for the use of Adacolumn and Cellsorba in the management of inflammatory bowel diseases. PMID- 18382086 TI - Advances in family-based association analysis. Introduction. PMID- 18382088 TI - Likelihood-based association analysis for nuclear families and unrelated subjects with missing genotype data. AB - Missing data occur in genetic association studies for several reasons including missing family members and uncertain haplotype phase. Maximum likelihood is a commonly used approach to accommodate missing data, but it can be difficult to apply to family-based association studies, because of possible loss of robustness to confounding by population stratification. Here a novel likelihood for nuclear families is proposed, in which distinct sets of association parameters are used to model the parental genotypes and the offspring genotypes. This approach is robust to population structure when the data are complete, and has only minor loss of robustness when there are missing data. It also allows a novel conditioning step that gives valid analysis for multiple offspring in the presence of linkage. Unrelated subjects are included by regarding them as the children of two missing parents. Simulations and theory indicate similar operating characteristics to TRANSMIT, but with no bias with missing data in the presence of linkage. In comparison with FBAT and PCPH, the proposed model is slightly less robust to population structure but has greater power to detect strong effects. In comparison to APL and MITDT, the model is more robust to stratification and can accommodate sibships of any size. The methods are implemented for binary and continuous traits in software, UNPHASED, available from the author. PMID- 18382089 TI - A family-based likelihood ratio test for general pedigree structures that allows for genotyping error and missing data. AB - The purpose of this work is the development of a family-based association test that allows for random genotyping errors and missing data and makes use of information on affected and unaffected pedigree members. We derive the conditional likelihood functions of the general nuclear family for the following scenarios: complete parental genotype data and no genotyping errors; only one genotyped parent and no genotyping errors; no parental genotype data and no genotyping errors; and no parental genotype data with genotyping errors. We find maximum likelihood estimates of the marker locus parameters, including the penetrances and population genotype frequencies under the null hypothesis that all penetrance values are equal and under the alternative hypothesis. We then compute the likelihood ratio test. We perform simulations to assess the adequacy of the central chi-square distribution approximation when the null hypothesis is true. We also perform simulations to compare the power of the TDT and this likelihood-based method. Finally, we apply our method to 23 SNPs genotyped in nuclear families from a recently published study of idiopathic scoliosis (IS). Our simulations suggest that this likelihood ratio test statistic follows a central chi-square distribution with 1 degree of freedom under the null hypothesis, even in the presence of missing data and genotyping errors. The power comparison shows that this likelihood ratio test is more powerful than the original TDT for the simulations considered. For the IS data, the marker rs7843033 shows the most significant evidence for our method (p = 0.0003), which is consistent with a previous report, which found rs7843033 to be the 2nd most significant TDTae p value among a set of 23 SNPs. PMID- 18382090 TI - Estimating relative risks from significant family-based association studies. AB - Two approaches are described to estimate relative risks from significant family based association studies. They can be used to obtain either point estimates or confidence regions. The approaches are evaluated by a simulation study and illustrated by application to a real data set. It is shown that both approaches largely reduce the bias in the relative risk estimates which can occur in case that the significant outcome of the study from which the relative risks are estimated is ignored. PMID- 18382091 TI - FBAT-SNP-PC: an approach for multiple markers and single trait in family-based association tests. AB - OBJECTIVE: Develop a new test for family-based association studies and continuous traits that incorporates power- enhancing techniques from two existing testing strategies. METHODS: The new procedure initiates with an extraction of the relevant information from the variability of the genotypes and an assessment of the approximate individual markers effects and their directions. This information is incorporated in the construction of the actual test statistic through a selection of a data-determined number of optimal linear combinations of the offspring genotypes which, in a power enhancing step, are consequently combined into a single degree of freedom test. We conduct a comparison simulation study in which the performance of the new test is contrasted with the test that is currently known to offer the highest overall power, FBAT-LC. RESULTS: The new test has an overall performance very similar to that of FBAT-LC but attains higher power in candidate genes with lower average pairwise correlations and moderate to high allele frequencies with large gains (up to 80%) for some of the analyzed genes possessing the above-mentioned characteristics. CONCLUSION: The new test is a promising tool for candidate gene studies with substantial power gains for genes that are characterized by SNPs with low mean pairwise correlation. PMID- 18382092 TI - The future is now - will the real disease gene please stand up? AB - The transmission/disequilibrium test (TDT) [Spielman et al.: Am J Hum Genet 1993;52:506-516] has been postulated as the future of gene mapping for complex diseases, provided one is able to genotype a dense enough map of markers across the genome. Risch and Merikangas [Science 1996;273:1516-1517] suggested a million marker screen in affected sibpair (ASP) families, demonstrating that the TDT is a more powerful test of linkage than traditional linkage tests based on allele sharing when there is also association between marker and disease alleles. While the future of genotyping has arrived, successes in family-based association studies have been modest. This is often attributed to excessive false positives in candidate gene studies. This problem is only exacerbated by the increasing numbers of whole genome association (WGA) screens. When applied in ASPs, the TDT statistic, which assumes transmissions to siblings are independent, is not expected to have a constant variance in the presence of variable linkage. This results in generally more extreme statistics, hence will further aggravate the problem of having a large number of positive results to sort through. So an important question is how many positive TDT results will show up on a chromosome containing a disease gene due only to linkage, and will they obfuscate the true disease gene location. To answer this question we combined theory and computer simulations. These studies show that in ASPs the normal version of the TDT statistic has a mean of 0 and a variance of 1 in unlinked regions, but has a variance larger than 1 in linked regions. In contrast, the pedigree disequilibrium test (PDT) statistic adjusts for correlation between siblings due to linkage and maintains a constant variance of 1 at unassociated markers irrespective of linkage. The TDT statistic is generally larger than the PDT statistic across linked regions. This is true for unassociated as well as associated markers. To compare the two tests we ranked both statistics at the disease locus, or an associated marker, among statistics at all other markers. The TDT did better job than PDT placing the score of the associated marker near the top. Though, strictly speaking, the TDT in ASPs should be interpreted as a test of linkage and not a test of association, there is a good chance that if a marker stands out, the marker is associated as well as linked. In conclusion, our results suggest that TDT is an effective screening tool for WGA studies, especially in multiplex families. PMID- 18382093 TI - The most important role of the mentor is to guide the mentee in finding own solutions to problems. An interview with Prof. Henri Sarles. Interview by Martin E. Fernandez-Zapico. AB - In this interview for Pancreatology, Professor Henri Sarles shares his life experiences as clinical expert and scientist in pancreatic research. He is a pancreatologist recognized worldwide for his contribution to the understanding of pancreatic diseases. In particular, he led the way in the pancreatitis field with the characterization of pathogenesis of chronic pancreatitis. PMID- 18382095 TI - Primers on molecular pathways--notch. AB - Notch, initially discovered and well characterized in Drosophila due to the notches apparent in the wing blades of mutant strains, and its signaling pathway play a key role in cell-cell communication, which involves gene regulation mechanisms that control multiple cell differentiation processes during embryonic and adult life, including timely cell lineage specification of both the endocrine and exocrine pancreas. This pathway serves as an excellent model signaling cascade for the regulation of the transition from normal ductal epithelium to metaplasia to cancer. In human pancreatic cancer, the change in epithelial differentiation programming is an early hallmark. Several of these changes, including activation of the Notch pathway, which are observed in pathological scenarios, are also seen during normal embryonic development of the pancreas. Notch receptors, ligands as well as downstream targets have been identified to be upregulated in preneoplastic lesions to invasive pancreatic cancers in humans and mice, suggesting that Notch signaling may be an early event leading to accumulation of undifferentiated precursor cells in pancreatic cancers. PMID- 18382087 TI - Review and evaluation of methods correcting for population stratification with a focus on underlying statistical principles. AB - When two or more populations have been separated by geographic or cultural boundaries for many generations, drift, spontaneous mutations, differential selection pressures and other factors may lead to allele frequency differences among populations. If these 'parental' populations subsequently come together and begin inter-mating, disequilibrium among linked markers may span a greater genetic distance than it typically does among populations under panmixia [see glossary]. This extended disequilibrium can make association studies highly effective and more economical than disequilibrium mapping in panmictic populations since less marker loci are needed to detect regions of the genome that harbor phenotype-influencing loci. However, under some circumstances, this process of intermating (as well as other processes) can produce disequilibrium between pairs of unlinked loci and thus create the possibility of confounding or spurious associations due to this population stratification. Accordingly, researchers are advised to employ valid statistical tests for linkage disequilibrium mapping allowing conduct of genetic association studies that control for such confounding. Many recent papers have addressed this need. We provide a comprehensive review of advances made in recent years in correcting for population stratification and then evaluate and synthesize these methods based on statistical principles such as (1) randomization, (2) conditioning on sufficient statistics, and (3) identifying whether the method is based on testing the genotype-phenotype covariance (conditional upon familial information) and/or testing departures of the marginal distribution from the expected genotypic frequencies. PMID- 18382096 TI - Assessment of the severity of acute pancreatitis: no room for complacency. AB - Assessing the severity of acute pancreatitis is an important initial step in the management of these patients. An ideal prognostic system or marker does not exist, and current approaches fall short of what is needed when dealing with individual patients. It is recommended that the evaluation of the performance of a particular prognostic system or marker should include the calculation of positive and negative likelihood ratios, derived from a combination of sensitivity and specificity. Knowing the pre-test probability of a particular endpoint and the likelihood ratios make it possible to derive the post-test probability for the presence or absence of that endpoint for the individual patient in that population. The change in probability from before to after the test gives an indication of the clinical usefulness of the test. Improving the performance of prognostic systems and markers remains a challenge and there is no room for complacency. There are two ways forward: either the prognostic systems and markers need to be used in a more intelligent way, with combinations, sequencing or artificial neural network techniques, or by the discovery of new markers that measure critical aspects of outcome determining pathophysiology. No lack of energy has been expended on the latter, while the former offers more immediate promise. PMID- 18382098 TI - Monocyte subsets and natural killer cells in acute pancreatitis. AB - BACKGROUND: Alteration of the immune system is one of the major mechanisms responsible for complications in severe acute pancreatitis (AP). The aim of our study was to provide a complex evaluation of peripheral blood monocyte subsets, natural killer cells (NK cells) and cytotoxic T lymphocytes in patients with different severity forms of AP. METHODS: 20 patients with mild AP and 15 with severe AP (S-AP) were included in our study. Peripheral blood mononuclear cells were studied on days 1-3, 5, 10 and 30, by means of flow cytometry. RESULTS: In peripheral blood of patients with pancreatitis, we found a marked increase in total monocyte count. In S-AP, circulating monocytes were significantly activated, which was presumed from increased expression of HLA-DR, CD54, CD69 and CD25. Concurrent increased expression of CD95 (FasR) may indicate enhanced susceptibility of these cells to apoptosis. In patients with S-AP, a dramatic depletion of circulating NK cells (CD16/56 and CD3- CD8+) was found along with a reduction of circulating CD3+ CD8+ lymphocytes (cytotoxic T lymphocytes). CONCLUSION: Our findings suggest profound disturbances of innate cellular immunity in patients with S-AP. PMID- 18382099 TI - Serous cystadenoma of the pancreas: clinical and pathological features in 33 patients. AB - AIM: To report the clinicopathological features of patients with serous cystadenomas of the pancreas. METHODS: Thirty-three cases of serous cystadenoma diagnosed between 1977 and 2006 were retrieved from the files of the Ohio State University Medical Center. Clinical data and microscopic slides were reviewed. RESULTS: The patients included 27 women and 6 men with an age range of 38-83 (mean 64.3) years. The clinical presentation included 13 patients with abdominal pain and 8 patients with abdominal mass; 9 tumors were found incidentally. Abdominal CT scans in 25 patients were interpreted as suspicious for carcinoma in 8 (32%), suspicious for serous cystadenoma in 8, neoplasm not otherwise specified in 8, and suspicious for a pseudocyst in 1. Only 7 patients underwent a preoperative biopsy, and 5 of these were diagnosed as having a serous cystadenoma. All but 2 of the patients underwent surgical resection of the tumor. The serous cystadenomas varied in size from 1.0 to up to 13 cm in maximum dimension, and all but one had a multicystic appearance. Of the 33 serous cystadenomas, 20 (61%) were located in the pancreatic tail, 4 (12%) in the pancreatic body, 4 in the pancreatic body and tail, and 5 (15%) in the head of the pancreas. Follow-up in 17 patients (median 3 years, range from 1 month to 11 years) showed no recurrence of serous cystadenomas. One patient had von Hippel Lindau syndrome, 4 patients had diabetes mellitus, 3 patients had metastatic cancer, and 2 patients had ovarian tumors. CONCLUSIONS: Serous cystadenoma is an uncommon neoplasm that can be confused with malignancy both clinically and radiologically; a correct diagnosis is important in order to provide an accurate prognosis. PMID- 18382097 TI - Pancreatic carcinogenesis. AB - Pancreatic cancer is an almost universally lethal disease. Research over the last two decades has shown that pancreatic cancer is fundamentally a genetic disease, caused by inherited germline and acquired somatic mutations in cancer-associated genes. Multiple alterations in genes that are important in pancreatic cancer progression have been identified, including tumor suppressor genes, oncogenes, and genome maintenance genes. Furthermore, the identification of noninvasive precursor lesions of pancreatic adenocarcinoma has led to the formulation of a multi-step progression model of pancreatic cancer and the subsequent identification of early and late genetic alterations culminating in invasive cancer. In addition, an increased understanding of the molecular basis of the disease has facilitated the identification of new drug targets enabling rational drug design. The elucidation of genetic alterations in combination with the development of high-throughput sensitive techniques should lead to the discovery of effective biomarkers for early detection of this malignancy. This review focuses mainly on the current knowledge about the molecular insights of the pathogenesis of pancreatic ductal adenocarcinoma. PMID- 18382100 TI - Pancreatogram findings for carcinoma in situ (CIS) of the pancreas seen on endoscopic retrograde cholangiopancreatography and postoperative pancreatography of resected specimens: can CIS be diagnosed preoperatively? AB - BACKGROUND/AIMS: From 1992 to 2003, 7 carcinomata in situ (CIS) were incidentally discovered during microscopical observation of resected materials for advanced carcinomas of peripancreatic organs, of which 4 had undergone endoscopic retrograde cholangiopancreatography (ERCP) or postoperative pancreatography of the resected specimen (POP). In addition, 7 of 79 invasive ductal carcinomata (IDC) of the pancreas were accompanied by CIS > or =2 cm long. A total of 11 patients were reviewed here for pancreatographic findings for CIS of the pancreas. METHODS: All resected pancreatobiliary materials were sliced serially at 5- to 8-mm intervals in a plane at right angles to the main pancreatic duct, referring to POP images. RESULTS: Irregularity (I), non-continuous narrowing (N), granular defects (G), and dilatation (D) were seen in 78, 67, 33 and 22% on ERCP, respectively, and in 90, 70, 60 and 40% on POP, respectively. CONCLUSIONS: I, N, G, and D are most important pancreatographic findings in ERCP and highly suggestive of CIS of the pancreas, so that whenever they are encountered, cytological and/or pathological examination of the pancreatic duct should be actively performed. PMID- 18382101 TI - Understanding the colonic complications of pancreatitis. AB - BACKGROUND: Colonic necrosis, fistula and stricture are infrequent but potentially lethal complications of pancreatitis. As any individual unit will have only limited experience, this study aims to provide a structured, systematic appraisal of published experience to identify any consistent trends and disease patterns that may help in practical management. METHODS: A computerized search of the MEDLINE databases for the period January 1950 through January 2006 yielded 43 articles. Pooled extracted data were examined for type of pancreatitis and colonic complications, method and time of diagnosis, treatment and outcome. RESULTS: 43 reports provided pooled data on 97 patients. Colonic complications were more frequent in severe disease, occurring in 15%. The principal presentations were necrosis, fistula and stricture. All episodes of colonic necrosis complicated severe acute pancreatitis, were diagnosed operatively, presented at a median of 25 (1-55) days into the episode and were associated with a mortality of 54%. In contrast, stricture presented at a median of 50 (10-270) days. Surgical resection without anastomosis is the mainstay of management of necrosis. Trial of conservative management in a stable patient with a fistula may facilitate spontaneous closure. CONCLUSIONS: This study highlights several consistent trends: preoperative diagnosis is difficult, colonic necrosis and fistula are rare complications principally of severe acute pancreatitis and they present either as ongoing abdominal sepsis or rectal bleeding. Surgical resection remains the mainstay of management. A high index of suspicion should be maintained in patients with severe acute pancreatitis, with ongoing sepsis and evidence of gastrointestinal blood loss. PMID- 18382102 TI - Percutaneous pancreatography for treatment of complicated pancreatic duct strictures. AB - BACKGROUND: Percutaneous transhepatic cholangiography was developed over 30 years ago for the diagnosis of biliary disorders. It has become an accepted interventional technique for management of biliary tract diseases, especially in patients who have failed attempted endoscopic retrograde pancreatography or have altered anatomy that makes the biliary tree endoscopically inaccessible. The correlative procedure, which we term 'percutaneous pancreatography' (PP), has only been described once in the literature. CASE PRESENTATIONS: We report the outcome of 4 patients undergoing PP for management of difficult pancreatic duct strictures. In all patients, PP was used to provide access and therapy of otherwise endoscopically impassable pancreatic duct strictures. PP-guided pancreatic stent duct placement was performed and allowed for subsequent successful endoscopic management of complex, benign pancreatic duct obstructions. CONCLUSIONS: PP is a useful modality for management of otherwise endoscopically impassable pancreatic duct strictures. PMID- 18382103 TI - Distribution of thyrotropin-releasing hormone (TRH) immunoreactivity in the brain of urodele amphibians. AB - To improve knowledge of the peptidergic systems in the brain of amphibians we have conducted a comparative analysis of the distribution of TRH immunoreactive cell bodies and fibers in three species of urodeles. Fiber labeling was observed in all main brain subdivisions suggesting different control functions for TRH in extrahypothalamic systems. However, as in other vertebrates, TRH neurons were abundant in the hypothalamic nuclei that presumably project to the median eminence and the neural lobe of the hypophysis. Considerable interspecies differences were noted mainly related to innervation of the olfactory and visual centers (thalamus and mesencephalic tectum) and the precise localization of immunoreactive cell bodies, which was assessed by double labeling with tyrosine hydroxylase. The comparison of the distribution of TRH immunoreactive neurons and fibers found in urodeles with those reported for other vertebrates, in particular with anamniotes, reveals a strong resemblance but also notable variations not only across vertebrate classes but also within the same class. In this respect, the virtual lack in urodeles of TRH innervation of the intermediate lobe of the hypophysis clearly contrasts with the innervation found in anurans. Therefore, the important role of skin color adaptation proposed for TRH in anurans on the basis of the direct innervation of the intermediate lobe is not applicable for urodeles. PMID- 18382105 TI - Muir-Torre syndrome. PMID- 18382104 TI - Lipomatous metaplasia after severe and chronic cutaneous inflammation. AB - A 69-year-old woman with a history of acute generalized exanthematic pustulosis (AGEP) caused by metamizole is described. Furthermore, she had suffered from an untreated psoriasis since the age of 20. After an adequate therapy of both psoriasis and AGEP, yellow-brownish, static, coalescing, lucent nodules on the thighs and upper arms became apparent. Histology of skin biopsies revealed a prominent band of mature adipocytes in the dermis. We diagnosed a lipomatous metaplasia of the dermis and hypothesize that this metaplasia occurred as a consequence of the severe and chronic inflammation of the skin. PMID- 18382106 TI - Introduction - microbubbles, ultrasound, and cerebrovascular disease. PMID- 18382107 TI - Clinical utility of contrast-enhanced ultrasound in neurosonology. AB - Extracranial and transcranial colour-coded duplex sonography (TCCD) are used routinely in clinical practice to assess physiologic and pathologic arterial blood flow to the brain. Such investigations are straightforward, safe, and cost effective, and can be repeated frequently for patient follow-up. In addition, the portability of extracranial colour-coded duplex sonography and TCCD provides the option of bedside investigations, particularly useful for evaluation of the critically ill patient. However, duplex ultrasound can be hampered by insufficient acoustic penetration, an unfavourable insonation angle, or by conditions of low or no flow. Echocontrast agents capable of surviving pulmonary and capillary transit have been developed, and such agents increase the Doppler signal and, therefore, the success rate of neurosonographic investigations. Approximately 20% of TCCD studies involve poor insonation conditions and, therefore, the use of echocontrast agents is of particular interest for this application. Levovist and SonoVue are the two agents currently approved for use in neurosonography. Such agents have a relatively long-lasting effect (up to 7 min), and can be administered as a single injection or, alternatively, a fractionated injection protocol can be used, depending on the quality of the window. PMID- 18382108 TI - Prognostic value of transcranial sonography in acute stroke patients. AB - Accurate assessment of stroke is critical for patient prognosis and selection of appropriate treatment regimens in order to optimize patient outcomes. Advanced neurosonologic techniques are straightforward, portable, and cost-effective, representing significant advantages over other noninvasive imaging modalities for monitoring of the hemodynamic status of acute ischemic stroke. Ultrasound findings acquired both early (<3 h from onset of stroke) and later (6-24 h after stroke) have demonstrated feasibility and validity for the detection of stenosis/occlusion of key intracranial structures, such as the middle cerebral artery, and for immediate and unambiguous indication of flow velocities, particularly when contrast enhancement is used. In addition, the target of thrombolysis can be identified and localized, and the success of therapy monitored, by transcranial ultrasound. Finally, transcranial ultrasound can be used to gauge the appropriateness of more complex and costly imaging studies, thereby optimizing utilization of health care resources. PMID- 18382109 TI - Contrast-enhanced ultrasound perfusion imaging in acute stroke patients. AB - The field of neurovascular ultrasound is expanding rapidly with exciting new applications. While ultrasound contrast agents were initially used to overcome insufficient transcranial bone windows for identification of the basal cerebral arteries, new-generation microbubbles in combination with very sensitive contrast specific ultrasound techniques now enable real-time visualization of stroke. This article will provide a review of recent and emerging developments in ultrasound technology and contrast-specific imaging techniques for evaluation of acute stroke patients. PMID- 18382110 TI - An in vivo investigation on the wound-healing effect of two medicinal herbs using an animal model with foot ulcer. AB - BACKGROUND/AIM: Over 190 million people in the world suffer from diabetes mellitus. Diabetics are 25 times more likely to have a leg amputated because of unhealing foot ulcers. Herbal medicine has been used in China to salvage the ulcerated limb. With the aim to study the efficacy of two commonly used herbs for ulcer healing, namely Radix Astragali and Radix Rehmanniae, a good animal model needs to be developed for a proper in vivo investigation. METHODS: Firstly, a diabetic animal model was established by streptozotocin injection. Then standard wounds were created on the feet of the diabetic rats. Digital photographs were taken and analyzed by a novel image analysis software. RESULTS: The average ulcer area in the Radix Rehmanniae treatment group was 11.45 mm2, which was significantly smaller than the 15.12 mm2 in the water treatment group (p = 0.04). Radix Astragali, on the other hand, was found to have no significant effect on ulcer shrinkage. CONCLUSION: Further investigation is needed for the identification of the active principles of Radix Rehmanniae. PMID- 18382111 TI - Heidelberg Phoneme Discrimination Test (HLAD): normative data for children of the third grade and correlation with spelling ability. AB - OBJECTIVE: The Heidelberg Phoneme Discrimination Test (HLAD), developed and standardized in 1998, is widely used in the differential diagnosis of dyslexia. Normative data have only been available for children of the 2nd and 4th grades, while norms for the 3rd grade are still missing. PATIENTS AND METHODS: We assessed three HLAD subtests [auditory phoneme discrimination, kinesthetic phoneme discrimination (repeating minimal pairs) and phoneme analysis] in 140 children of the 3rd grade from eight elementary schools. Writing capacity was tested via DRT3. RESULTS: Comparing children of the 2nd, 3rd and 4th grades, we found a continuing increase in phoneme discrimination capacity with age. This increase was especially evident for the task of auditory comparison. For the 3rd grade, the correlation between HLAD and writing test (qualitative analysis) was 0.55, and 0.36 between HLAD and writing (quantitative analysis). The correlation with writing tasks was highest in the 2nd grade. CONCLUSION: The steady increase in phoneme discrimination capacity from the 2nd to 4th grade may indicate maturation and learning effects at least until the age of 10 years. PMID- 18382112 TI - Vitamin D: a candidate for the environmental effect in multiple sclerosis - observations from Norway. AB - Multiple sclerosis (MS) is a chronic disease of the central nervous system, pathologically characterized by inflammation, demyelination, and axonal damage, presumably auto-immune in nature. Complex interactions between genetic susceptibility and environmental factors, such as vitamin D status and primary Epstein-Barr virus infection in adolescence or later in life, probably determine the MS risk. Norway at a latitude 58-71 degrees N is a discrete exception to the hypothesis that solar UV radiation exposure, mediated by vitamin D, coheres with the latitude gradient seen for MS prevalence. Where UV radiation exposure is low in Norway,vitamin D sufficiency is maintained through a traditional diet providing vitamin D as well as marine omega-3 polyunsaturated fatty acids. This observation supports an environmental interaction between diet and latitude, with vitamin D as the common mediator. The potential roles of vitamin D, other environmental exposures, and genes in the complex aetiology of MS are discussed in this review. PMID- 18382113 TI - Incidence of multiple sclerosis in the urban area of Erfurt, Thuringia, Germany. AB - European data on the epidemiology of multiple sclerosis (MS) reveal a continuous increase in the incidence of MS, which does not as yet seem to have reached a plateau at any point in time. No epidemiological data are available from Thuringia, which belonged to the former German Democratic Republic and which was politically separated from the neighbouring epidemiologically well-known areas of Hesse and Lower Saxony. We determined the incidence of MS in the urban area of Erfurt in Thuringia, by prospective identification of patients, with a diagnosis of definite or probable MS between 1998 and 2002. The crude annual incidence was 8.0:100,000, which is the highest rate ever reported for Germany. Our data, showing a high proportion of females with a late disease onset, suggests an increase in female cases with a benign form of MS resulting in an increase in MS incidence. An impact of changing diagnostic procedures inflating the rate has still to be taken into account. Until now it is partly up to speculation whether the observed increase in incidence may be related to emerging new cases, possibly from the pool of prior subclinical MS cases. PMID- 18382114 TI - Incidence of dementia, Alzheimer disease, and vascular dementia in a Japanese population: Radiation Effects Research Foundation adult health study. AB - OBJECTIVE: To determine the age-, sex-, and subtype-specific incidence of dementia and to assess the effect of education level on the incidence in a Japanese population. METHODS: 2,286 dementia-free subjects, aged > or =60 years, were followed for 5.9 years through biennial two-phase examinations. RESULTS: 206 cases of dementia were newly diagnosed based on DSM IV. The incidence per 1,000 person-years was 12.0 for men and 16.6 for women. Based on NINCDS-ADRDA criteria, 80 cases of probable Alzheimer disease (AD) and 50 cases of possible AD were diagnosed. Based on NINDS-AIREN criteria, 36 cases of probable vascular dementia (VaD) and 40 cases of possible VaD were diagnosed. Age and education showed the most statistically significant effects for all dementia. Probable AD showed the most remarkable increase with age and decreased with increasing education level (p = 0.001). Probable VaD showed significant effects of sex (p = 0.033) and sex age interaction (p = 0.048), but not education (p = 0.26). CONCLUSION: AD was the predominant type of dementia in this recent incidence study conducted in Japan, suggesting a reduction in VaD and an increase in AD. Age, sex, and education effects differed by dementia subtype. PMID- 18382115 TI - Dietary epidemiology of essential tremor: meat consumption and meat cooking practices. AB - BACKGROUND/AIM: Harmane [1-methyl-9H-pyrido(3,4-b)indole] is a tremor-producing neurotoxin. Blood harmane concentrations are elevated in essential tremor (ET) patients for unclear reasons. Potential mechanisms include increased dietary harmane intake (especially through well-cooked meat) or genetic-metabolic factors. We tested the hypothesis that meat consumption and level of meat doneness are higher in ET cases than in controls. METHODS: Detailed data were collected using the Lawrence Livermore National Laboratory Meat Questionnaire. RESULTS: Total current meat consumption was greater in men with than without ET (135.3 +/- 71.1 vs. 110.6 +/- 80.4 g/day, p = 0.03) but not in women with versus without ET (80.6 +/- 50.0 vs. 79.3 +/- 51.0 g/day, p = 0.76). In an adjusted logistic regression analysis in males, higher total current meat consumption was associated with ET (OR = 1.006, p = 0.04, i.e., with 10 additional g/day of meat, odds of ET increased by 6%). Male cases had higher odds of being in the highest than lowest quartile of total current meat consumption (adjusted OR = 21.36, p = 0.001). Meat doneness level was similar in cases and controls. CONCLUSION: This study provides evidence of a dietary difference between male ET cases and male controls. The etiological ramifications of these results warrant additional investigation. PMID- 18382116 TI - Prevalence and incidence of multiple sclerosis in western Greece: a 23-year survey. AB - BACKGROUND: The frequency of multiple sclerosis (MS) in Greece remains speculative, as data from many parts are still lacking. OBJECTIVE: To estimate trends in MS prevalence and annual incidence in western Greece from January 1, 1984 to December 31, 2006. METHODS: Patients were identified from the patient records of the Department of Neurology at Patras University Hospital in Rion Patras. Only patients with a definite MS diagnosis according to Poser's criteria and retrospective application of the McDonald's criteria were included. We calculated age- and sex-specific prevalence rates for patients living in the study area on December 31, 2006. Annual incidence rates were calculated for the period 1984-2006. RESULTS: The crude prevalence rate of definite MS cases increased significantly in 23 years from 10.1/100,000 recorded in northeastern Greece in 1984 to 119.61/100,000 on December 31, 2006 in western Greece for the 780 cases still alive. The mean annual incidence rate increased from 2.71/100,000 recorded during the period 1984-1989 to 10.73/100,000 in the 5-year period of 2002-2006. CONCLUSION: The prevalence rates were higher than expected, but closer than in previous surveys conducted in Greece to those reported recently in Sicily and Istanbul. These findings place the area in the high-risk zone. PMID- 18382117 TI - Reduced maximal oxygen consumption and overproduction of proinflammatory cytokines in athletes. AB - OBJECTIVE: It was the aim of this study to evaluate whether chronic pain in athletes is related to performance, measured by the maximum oxygen consumption and production of hormones and cytokines. METHODS: Fifty-five athletes with a mean age of 31.9 +/- 4.2 years engaged in regular competition and showing no symptoms of acute inflammation, particularly fever, were studied. They were divided into 2 subgroups according to the occurrence of pain. Plasma concentrations of adrenaline, noradrenaline, cortisol, prolactin, growth hormone and dopamine were measured by radioimmunoassay, and the production of the cytokines interleukin (IL)-1, IL-2, IL-4, IL-6, tumor necrosis factor-alpha, interferon-alpha and prostaglandin E(2) by whole-blood culture. Maximal oxygen consumption was determined during an incremental treadmill test. RESULTS: There was no change in the concentration of stress hormones, but the athletes with chronic pain showed a reduction in maximum oxygen consumption (22%) and total consumption at the anaerobic threshold (25%), as well as increased cytokine production. Increases of 2.7-, 8.1-, 1.7- and 3.7-fold were observed for IL-1, IL 2, tumor necrosis factor-alpha and interferon-alpha, respectively. CONCLUSIONS: Our data show that athletes with chronic pain have enhanced production of proinflammatory cytokines and lipid mediators and reduced performance in the ergospirometric test. PMID- 18382118 TI - Temperamental activity and epidermal growth factor A61G polymorphism in Finnish adults. AB - BACKGROUND/AIMS: Epidermal growth factor (EGF) has widespread effects on the developing and mature nervous system. This study examined whether genetic differences in the EGF predict differences in the self-reported level of activity temperament in adults. METHODS: Two hundred and ninety-two men and women from the population-based Cardiovascular Risk in Young Finns Study were genotyped for the functional EGF gene A61G polymorphisms, and their temperamental activity was assessed 3 times in 1992, 1997 and 2001. RESULTS: The EGF gene predicted temperamental activity (the mean of 3 assessments, p = 0.007), with the G/G genotype being associated with the highest level of activity. Additional analyses in separate years indicated that the association was robust across different measurements of activity (p < or = 0.05 in 1992 and 1997, p = 0.008 in 2001). CONCLUSION: These results suggest that polymorphic variation in the EGF A61 gene may be one of the factors underlying the temperament dimension of activity. PMID- 18382119 TI - Increased plasma leptin in antipsychotic-naive females with schizophrenia, but not in males. AB - BACKGROUND: Metabolic abnormalities, including insulin resistance and increased leptin levels, were noted in patients with schizophrenia who had received antipsychotics. In this study, we examined the leptin levels of antipsychotic naive schizophrenic patients. METHOD: Seventeen antipsychotic-naive patients with schizophrenia and 16 sex-, age- and body mass index (BMI)-matched subjects were recruited from the psychiatric outpatient clinic and community, respectively. Serum leptin levels of the healthy controls and antipsychotic-naive patients were compared. The relationships between leptin level and gender, Positive and Negative Syndrome Scale score, and duration of illness were analyzed by Spearman's correlation. RESULTS: Compared with sex-, age- and BMI-matched subjects, the female schizophrenic patients had elevated serum leptin levels. In the male patients, the results of this preliminary study also revealed a positive correlation between leptin levels and the Aggression Risk Profile subscale of the PANSS. CONCLUSION: This preliminary study suggests possible gender-specific leptin dysregulation in antipsychotic-naive schizophrenic patients. PMID- 18382120 TI - Cerebellar posterior superior vermis and cognitive cluster scores in drug-naive patients with first-episode schizophrenia. AB - Previously, we performed an MRI study that revealed smaller volumes of the subregions of the cerebellar vermis in men and women with chronic schizophrenia. An issue that arose from that study was whether similar structural changes in the cerebellum are found in patients with first-episode schizophrenia. In the present study, MRI scans were acquired from 14 drug-naive patients with first-episode schizophrenia and 16 healthy subjects, and used to measure the volumes of their cerebellar subregions. Positive symptom, negative symptom and cognitive cluster scores were attained using the Positive and Negative Syndrome Scale. Patients with first-episode schizophrenia had reduced volumes of the anterior vermis and posterior superior vermis compared with healthy subjects. We confirmed that there was a volume reduction of the cerebellar vermis in drug-naive patients with first episode schizophrenia. Smaller volumes of the posterior superior vermis were associated with worse cognitive cluster scores in patients with first-episode schizophrenia. PMID- 18382121 TI - Structural insights into the functions of the large ribosomal subunit, a major antibiotic target. PMID- 18382122 TI - Intersubjectivity, affective neuroscience, and the neurobiology of autistic spectrum disorders: a systematic review. AB - Intersubjectivity is an approach to the study of social interaction viewed from a perspective which rejects the view that reducing any such analysis to study at the level of the individual is adequate to address the issues of social functioning. It also stresses the view that social processes cannot be reduced to cognitive ones - most of the important questions in the study of developmental psychopathology deal with issues which have commonality with many other species and are patent well before the ontological emergence of 'cognitive' abilities. In this paper we review the evidence in this area, and discuss a range of issues relevant to autistic spectrum disorders. We focus in particular on social interaction; the role of the Intrinsic Motive Formation and recent work on mirror neurons in autism; genetic and teratogenic factors in the genesis of autism; and the role of a number of biological factors in pathogenesis - tryptophan; vitamin B12; sterol metabolism; glutamate and GABA; and the Fragile-X expansion. PMID- 18382123 TI - Managing the conflict between individual needs and group interests--ethical leadership in health care organizations. AB - This paper derives from a grounded theory study of how Medical Directors working within the UK National Health Service manage the moral quandaries that they encounter as leaders of health care organizations. The reason health care organizations exist is to provide better care for individuals through providing shared resources for groups of people. This creates a paradox at the heart of health care organization, because serving the interests of groups sometimes runs counter to serving the needs of individuals. The paradox presents ethical dilemmas at every level of the organization, from the boardroom to the bedside. Medical Directors experience these organizational ethical dilemmas most acutely by virtue of their position in the organization. As doctors, their professional ethic obliges them to put the interests of individual patients first. As executive directors, their role is to help secure the delivery of services that meet the needs of the whole patient population. What should they do when the interests of groups of patients, and of individual patients, appear to conflict? The first task of an ethical healthcare organization is to secure the trust of patients, and two examples of medical ethical leadership are discussed against this background. These examples suggest that conflict between individual and population needs is integral to health care organization, so dilemmas addressed at one level of the organization inevitably re-emerge in altered form at other levels. Finally, analysis of the ethical activity that Medical Directors have described affords insight into the interpersonal components of ethical skill and knowledge. PMID- 18382124 TI - Lowering the premature birth rate: what the U.S. experience means for Japan. AB - Premature birth rate and low birth weight rate are increasing in industrialized countries including USA and Japan. The Infant mortality rate (IMR) is three times and 50-75 times greater for infants born at 32-36 weeks and <32 weeks respectively than term-born counterparts. In the U.S., the IMR is greater than in Japan particularly among black infants and simply the "lower socioeconomic class" is not the answer. Premature birth is heterogeneous in origin and idiopathic in 70% of the cases. Increased utilization of assisted reproductive technology only accounts for a part of the recent trend. Evidence suggests environmental factors play a significant role, and genetic-environmental interaction is plausible. A chronic psychosocial stress of pregnant women has been postulated to be modifying the endocrine milieu thereby influencing pregnancy outcomes. In a preliminary observation in St. Louis, homeless pregnant women with high behavioral and social risks, when accommodated in a shelter home designed for these women, produced significantly less numbers of premature and low birth weight infants as compared with the general population. Furthermore, in a randomized controlled study in Washington DC, psychobehavioral intervention specifically targeting smoking (primary and secondary), intimate partner violence (IPV), and depression among black pregnant women significantly decreased the rate of miscarriage and low birth weight. These reports may have significant implication to the Japanese situation. Increasing number of Japanese women at reproductive age are exposed to smoking, may have underling psychosocial stress and may suffer from subclinical depression and/or from IPV. Detailed epidemiological studies of women before and during the reproductive age with regard to risk factors can lead to an effective intervention strategy against premature birth in Japan. PMID- 18382125 TI - Task-and phase-related changes in cortico-muscular coherence. AB - Cortico-muscular coherence was compared during ramp-and-hold isometric and quasi isotonic contraction of the ankle joint in human subjects. EEG was recorded from the leg area of the motor cortex. EMG was recorded from the tibialis anterior (TA) muscle. The subjects were requested to maintain a steady low level of dorsiflexion and at intervals of 10 s to increase the contraction level within 1 s, maintain this level for 4 s and then decrease the level of contraction again within another 1 s. In seven subjects coherence in the 15-35 Hz frequency band was seen between EEG and TA EMG during low-level tonic dorsiflexion. In all subjects coherence disappeared during the ramp phase for both isometric and quasi isotonic contraction. Coherence at other frequencies was also not observed in any of the subjects during the ramp phase. During the hold phase at the stronger level of contraction coherence reappeared quickly and had the same size as at the low level of contraction. However, a significantly larger level of coherence was found during quasi-isotonic than during the isometric contraction. This demonstrates that cortico-muscular coherence in the 15-35 Hz frequency band is phase- and task-related. The decrease in 15-35 Hz coherence during the ramp phase may be related to event-related desynchronization of EEG activity. The larger level of coherence during quasi-isotonic contraction may reflect a higher demand of precise control of the joint position. It may also reflect a greater need for binding of functionally related cortical pyramidal tract neurons. PMID- 18382126 TI - Early detection of melanoma progression by quantitative real-time RT-PCR analysis for multiple melanoma markers. AB - Standard screening of melanoma patients is a useful tool for predicting outcome of patients, however, an instant methodology for exact detection of subclinical disease or monitoring treatment response is under investigation. Detection of circulating melanoma cells is, therefore, a possible novel promising staging method. However, inconsistent data on method sensitivity and on the predicted patient outcome has been shown repeatedly. Recently, a multimarker real-time RT PCR methodology for quantification of five melanoma markers Melan-A, gp 100, MAGE 3, MIA and tyrosinase was described by our group. In the current prospective trial, blood specimens of 65 patients with AJCC stage IIB-III cutaneous melanoma after surgery were periodically examined. In the above group, 27 % of subjects relapsed during the study. Prior to the disease progression we could observe a statistically significant tumor marker elevation in previous 0 to 9 months in all patients with clinical relapse. MAGE-3 became the most sensitive progression marker. During progression, three concordant positive markers were seen in 39 % of patients, followed by two concordant positive markers in 28 % and 1 marker in 33 %. This study supports the use of a multimarker real-time RT-PCR as a disease progression predictor. The dynamic assessment of serially obtained blood specimens represents a useful method for early metastasis detection and treatment response of melanoma patients. PMID- 18382127 TI - CARPs enhance p53 turnover by degrading 14-3-3sigma and stabilizing MDM2. AB - CARP1 and CARP2 proteins (CARPs) are E3 ligases that target p53 as well as phospho-p53 for degradation. Because MDM2 is a critical regulator of p53 turnover, we investigated and found that CARPs associate with MDM2. We provide evidence that CARPs stabilize MDM2 by inhibiting MDM2 self-ubiquitination. CARPs together with MDM2 enhance p53 degradation, thereby inhibiting p53-mediated cell death. CARP protein levels correlate with MDM2 levels including under hypoxia where both are reduced. CARP2 was found to target 14-3-3sigma for degradation, leading to MDM2 stabilization. MDMX, a homolog of MDM2, is not absolutely required for MDM2 stabilization by CARPs, although overexpression of CARP2 enhances MDM2/MDMX interaction. Taken together, our study identifies novel mechanisms by which CARP proteins regulate the p53 signaling pathway. PMID- 18382128 TI - Progress in dendritic cell immunotherapy: elucidating the enigma of Th-1 polarization. AB - The goal of harnessing the immune system to effectively eradicate neoplastic disease will require the generation of robust Th-1 type immunity and durable immunological memory against the antigenic repertoire that differentiates normal self from neoplastic self. While the literature presents a very mixed picture as to the requirement of T-cell help for the generation of both primary and memory CTL responses, there appears to be a general consensus that CD4(+) T-cell help will be required to generate durable responses against self cancer antigens that are devoid of foreign PAMPs and for which high-affinity T-cell clones have been deleted as a consequence of thymic selection. Here we comment briefly upon the characterization of an emerging regulatory pathway that enhances Th-1 polarization and CD8(+) CTL responses by a mechanism dependent upon CD40L mediated T-cell help. Further, we speculate that the full elucidation of this mechanism might be generally useful in answering some unresolved questions regarding the initiation of Th-1 polarized responses. PMID- 18382129 TI - Immunogenicity of a thermally inactivated rotavirus vaccine in mice. AB - Current approaches to the prevention of severe rotavirus diarrhea and deaths in children have all been through the use of live oral vaccines. To develop a safe and effective inactivated rotavirus vaccine (IRV), a new simple, rapid and robust method for the inactivation is critical and essential because chemical inactivation commonly used for a number of killed vaccines has been a challenge and problematic for rotavirus. We have examined an array of thermal conditions and demonstrated that purified YK-1 rotavirus in diluent buffer can be completely inactivated by heat treatment, as evidenced by the lack of virus growth in two successive passages in cell culture. Unlike chemical treatment that often causes physical and biochemical damages to viruses, thermally inactivated rotavirus particles maintained their structural, biochemical and antigenic integrity. A two dose intramuscular administration of thermally inactivated YK-1 rotavirus without adjuvant resulted in high titers of total and neutralizing antibody in serum of mice. Adjuvant Al(OH)(3) further led to enhanced antibody titers and also dramatically lowered the amount of antigens in the vaccine formulation. Our results demonstrate the potential of heat inactivation as a novel approach to the manufacture of a safe and efficacious parenteral rotavirus vaccine, which should serve as an important addition to and back up for live oral rotavirus vaccine in children. PMID- 18382130 TI - Towards prediction of degenerate CTL epitope recognition. AB - The cellular immune system is characterized by flexibility with respect to epitope recognition at the level of peptide binding to HLA molecules and HLA peptide complexes to T-cell receptors (TCRs). For epitopes recognized by cytotoxic T-lymphocytes (CTLs), amino acid substitutions at different positions have varying impact on recognition. By analyzing the frequencies of specific amino acid substitutions at each position in conjunction with HLA-peptide binding and immune-response data, we have developed new methods to predict cross-reactive recognition of epitope variants by CTLs. We derived position-specific substitution matrices (EPSSMs) through the analysis of known HLA ligands and achieved relatively accurate prediction of detrimental and tolerated amino acid substitutions. Initial analysis of amino acid substitutions in CTL epitopes with degenerate recognition showed strong position-specific preferences. This first systematic analysis further suggested that spatial constraint may be the major molecular factor determining the degenerate epitope recognition. As the data cumulates, we anticipate that eventually EPSSMs will be available for prediction of degenerate T-cell epitope recognition. PMID- 18382131 TI - A call to cellular & humoral arms: enlisting cognate T cell help to develop broad spectrum vaccines against influenza A. AB - Influenza A is an important cause of morbidity and mortality worldwide. In the United States alone influenza kills 30,000 to 50,000 people in a non-epidemic year and significantly more in an acute epidemic.(1) An emerging pandemic influenza virus, such as H5N1, could have a devastating economic and social impact. The Surgeon General estimates that at least 43 million Americans, especially those younger than 1 and older than 60, are at risk of death from influenza. Antigenically distinct influenza virus strains emerge regularly, mandating changes in influenza vaccine antigenic composition. Consequently, the immunity engendered by the conventional influenza vaccines is relevant only for a short time. However, by incorporating conserved influenza T cell epitopes, it may be possible to develop more immunogenic, broader-spectrum vaccines that may be efficacious over a longer period. This review summarizes the critical components of effective influenza vaccines, a rational vaccine design approach, and the pertinent influenza immunology. PMID- 18382132 TI - Evaluation of two yellow fever vaccines for routine immunization programs in Argentina. AB - Although highly effective vaccines have been available for almost 70 years, an estimated 200,000 cases of YF, including 30,000 deaths, still occur annually. This study evaluated the safety of two yellow fever (YF) vaccines [Stamaril and Vacina Contra Febre Amarela (VCFA)]. A total of 2,514 subjects were randomized equally to receive Stamaril or VCFA. Immediate reactions occurring within 30 minutes after vaccination, and solicited local and systemic reactions occurring within eight days, were monitored. Unsolicited local, systemic adverse events and serious adverse events (SAE) were recorded for 21 days after vaccination. Solicited local and systemic adverse reactions were reported by 15.3-17.6% and 30.4-31.6% of the Stamaril and VCFA groups, respectively. Only 56 of the 2,514 study subjects (2.2%) reported a severe solicited adverse reaction, 25 in the Stamaril group (1.99%) and 31 in the VFCA group (2.49%), (p=0.403). Ten subjects (0.8%) in each group reported at least one severe solicited local reaction (p = 0.988). A total of 18 Stamaril subjects (1.43%) and 21 VCFA subjects (1.68%) reported at least one severe solicited systemic reaction (p = 0.617) One SAE considered related to vaccination occurred, polymyalgia in the VCFA group. No immediate reactions to vaccination were seen. Vaccine-related unsolicited events were infrequent, 1.4% in the Stamaril group and 2.0% VCFA group, generally of mild or moderate intensity. We conclude that the safety profiles of Stamaril and VCFA support routine vaccination to prevent YF in residents of and travelers to endemic areas of South America and Africa. PMID- 18382133 TI - Preclinical profiling of the immunogenicity of a two-component subunit malaria vaccine candidate based on virosome technology. AB - Presentation of synthetic peptides on immunopotentiating reconstituted influenza virosomes is a promising technology for subunit vaccine development. An optimized virosomally delivered peptide representing 5 NPNA repeats of P. falciparum circumsporozoite protein is highly immunogenic in mice. Antibodies against this peptide (UK-39) inhibit sporozoite invasion of human hepatocytes. A second peptide (AMA49-C1) based on domain III of apical membrane antigen 1, induces antibodies that inhibit blood-stage parasite growth in vitro. Here we show a detailed pre-clinical profiling of these virosomally formulated peptides alone and in combination in mice and rabbits. Two immunizations with virosomally formulated UK-39 or AMA49-C1 were enough to elicit high titers of parasite cross reactive antibodies in both species. A low dose of 10 microg UK-39 was enough to induce maximal titers in rabbits. Higher doses of peptide did not increase antibody titers. In contrast, AMA49-C1 induced higher antibody titers with 25 and 50 microg peptide. Combination of UK-39 and AMA49- C1 on separate virosomes did not have any negative effect on anti-peptide antibody titers in mice or rabbits. No MHC restriction was observed in the development of humoral responses in outbred rabbits with different immunogenetic backgrounds. All vaccine formulations were safe in toxicity studies in rabbits and rats. Taken together, low amounts of synthetic peptides delivered on virosomes induced high antibody titers in mice and rabbits. Moreover, different peptides could be combined without interfering with individual anti-peptide responses, augmenting the value of this system for the development of a multivalent malaria vaccine. PMID- 18382134 TI - Self-reported adverse health events following smallpox vaccination in a large prospective study of US military service members. AB - In December 2002, the Department of Defense re-instituted smallpox vaccination for US military forces following growing concerns that smallpox might be employed as a bioterrorist weapon. More than one million service members have been given the smallpox vaccine since 2002, although there have been concerns about the safety of the vaccine. Using a large self-reported prospective database, this analysis investigated a wide variety of self-reported health outcomes and possible association with smallpox vaccination. After confirming self-reported vaccination history with electronic vaccine data, 40,472 individuals were included in the analyses, 8,793 of whom received the smallpox vaccine and 31,679 who did not. No significant adverse associations between smallpox vaccination and self-reported health outcomes, including mental and physical functioning, cardiovascular diseases, and autoimmune disorders, were found. These findings complement studies that utilize other data sources, such as electronic hospitalization records, and may be reassuring to health care providers and those who receive the smallpox vaccination. PMID- 18382135 TI - Formulation and characterization of a ten-peptide single-vial vaccine, EP-2101, designed to induce cytotoxic T-lymphocyte responses for cancer immunotherapy. AB - Effective vaccines that mediate clinical responses in cancer patients may require generation of broadly specific cytotoxic T lymphocytes (CTL) directed against multiple epitopes and tumor-associated antigens (TAA). Pursuant to this goal we developed a synthetic peptide vaccine, EP-2101, composed of 10 synthetic peptide epitopes and formulated in Montanide ISA 51 adjuvant. Nine of the HLA-A*0201 restricted CTL epitopes were derived from five well-characterized TAA. The universal HLA-DR binding epitope PADRE was also included for T-cell help. Herein we describe studies on the formulation and characterization of the EP-2101 vaccine which supports generation of a sterile single-vial emulsion using standardized processes. The physicochemical properties of the peptides were highly disparate and as such, solubilization studies were required to identify a process which supported sterile filtration of the EP-2101 peptide mix. A homogenization-based formulation process with Montanide ISA 51 and 0.5 mg/ml of each peptide was developed to generate a water-in-oil emulsion. Physical studies indicated the vaccine emulsion to be stable, with little change in visual appearance, viscosity and water droplet size for at least three months. The physical stability of individual peptides in the vaccine emulsion was demonstrated using HPLC and immunogenicity of the vaccine formulation was confirmed in HLA-A*0201/K(b) transgenic mice where T-cell responses could be induced to all epitopes in EP-2101 following vaccination. Our study process is scalable for production of approximately 1.5 liters of potent experimental vaccine for preclinical animal toxicity and phase 1 clinical testing in patients with breast, colon or lung cancer. PMID- 18382136 TI - Changes in DT vaccine frequency and indications for use following introduction of DTaP vaccine. AB - Our population-based study among HMO members under seven years of age in 1993 2000 showed that frequency of diphtheria and tetanus toxoid vaccine (DT) use declined significantly following the introduction of acellular pertussis containing (DTaP) vaccine. We also observed changes in indications for DT following the transition to DTaP among children under two years of age; notably, a decline in the proportion of children receiving DT due to a reported prior vaccine reaction and an increase in the proportion of children receiving DT due to parental request and a history of pertussis. PMID- 18382137 TI - The potential cost-effectiveness of vaccination against herpes zoster and post herpetic neuralgia. AB - A clinical trial has shown that a live-attenuated varicella-zoster virus vaccine is effective against herpes zoster (HZ) and post-herpetic neuralgia (PHN). The aim of this study was to examine the cost-effectiveness of vaccination against HZ and PHN in Canada. A cohort model was developed to estimate the burden of HZ and the cost-effectiveness of HZ vaccination, using Canadian population-based data. Different ages at vaccination were examined and probabilistic sensitivity analysis was performed. The economic evaluation was conducted from the ministry of health perspective and 5% discounting was used for costs and benefits. In Canada (population = 30 million), we estimate that each year there are 130,000 new cases of HZ, 17,000 cases of PHN and 20 deaths. Most of the pain and suffering is borne by adults over the age of 60 years and is due to PHN. Vaccinating 65-year-olds (HZ efficacy = 63%, PHN efficacy = 67%, no waning, cost/course = $150) is estimated to cost $33,000 per QALY-gained (90% CrI: 19,000 63,000). Assuming the cost per course of HZ vaccination is $150, probabilistic sensitivity analysis suggest that vaccinating between 65 and 75 years of age will likely yield cost-effectiveness ratios below $40,000 per Quality-Adjusted Life Year (QALY) gained, while vaccinating adults older than 75 years will yield ratios less than $70,000 per QALY-gained. These results are most sensitive to the duration of vaccine protection and the cost of vaccination. In conclusion, results suggest that vaccinating adults between the ages of 65 and 75 years is likely to be cost-effective and thus to be a judicious use of scarce health care resources. PMID- 18382138 TI - Novel vaccines and adjuvant systems: the utility of animal models for predicting immunogenicity in humans. AB - Animal models are essential for acquiring safety, immunogenicity and efficacy data to support the development of novel vaccines. However, extrapolating such results to designing human trials is challenging due to species-specific differences in responses to antigens, adjuvants and pathogens. As well, most early vaccine work is conducted with in-bred mouse strains which may fail to uncover issues that could arise later in out-bred populations. Unlike drugs designed to be delivered systemically, vaccines work within a somewhat localized space, so allometric dose scaling to account for body size differences is not necessarily relevant. Comparison of immune responses and correlates of protection with a given antigen show widely variable results between animals and humans, even where protective immunity against challenge with the same pathogen can be studied. For adjuvants, it is possible to compare enhancement of immunogenicity compared to a non-adjuvant control vaccine. While some novel adjuvants provide similar levels of enhancement between species, others do not. It is also important to recognize the inter-relationship between antigens and adjuvants, since one can compensate for the other, masking particular effects. Despite all the limitations, animal immunogenicity and efficacy studies form an important part of pre-clinical development for novel vaccines, but considerable prudence is required when using and extrapolating results. PMID- 18382139 TI - Development of a Chlamydia trachomatis bacterial ghost vaccine to fight human blindness. AB - Trachoma is the world's leading cause of preventable disease and the third most common cause of blindness after cataract and glaucoma, affecting an estimated 84 million people and leaving 590 million at risk. As a crippling disease, trachoma causes an enormous loss of productivity and constitutes a major socioeconomic burden. Although antibiotics are effective in treating active cases of the illness, they do not prevent re-infection, which occurs with high frequency in susceptible populations. Also, once infection and pathology are established, treatment may be less effective. Another major public health challenge posed by trachoma is that a large number of infected individuals are asymptomatic and readily infect those with whom they interact. Thus, an inexpensive and easy to deliver vaccine for trachoma would be highly effective in reducing the devastation caused by this disease. Development of an effective vaccine for controlling and preventing trachoma will require an understanding of the complex immunological mechanisms that occur during infection, identifying those antigens that elicit a protective immune response and designing effective vaccine delivery systems. Significant progress has been made in the delineation of the immune correlates of protection that will form the basis of vaccine evaluation. Recent advances in chlamydial genomics and proteomics has identified a number of protective antigens or epitopes that when appropriately delivered will produce an efficacious vaccine. The challenge at this time is the development of effective methods for vaccine delivery. We have developed an effective bacterial ghost (BG) delivery system possessing intrinsic adjuvant properties and capable of simultaneously delivering multiple antigens to the immune system. Such a flexible delivery system can produce an effective vaccine that will prevent the development of trachomatous conjunctivitis and blindness. The safety and relatively cheap production cost of BG-based vaccines offer a technological and manufacturing advantage for a vaccine needed on a global scale. PMID- 18382140 TI - Smallpox vaccination is not associated with infertility in a healthy young adult population. AB - Concerns exist regarding reproductive health, including potential infertility, among young adults with military-related occupational exposures. This study evaluated infertility diagnoses in a large population of healthy young adults in relation to prior smallpox vaccination. Using a retrospective cohort design, the population consisted of United States military members eligible for smallpox vaccination in 2003-2004 who had electronic health care utilization records available through at least December 2005. Multivariable logistic regression models were applied to evaluate infertility among male and female populations separately. Among 253,973 men and 44,332 women included in these analyses, the adjusted odds of infertility diagnoses in those with prior smallpox vaccination were 0.94 [95% confidence interval (CI), 0.83-1.06] and 1.10 (95% CI, 0.94-1.28), respectively. Therefore, no association was found between smallpox vaccination and subsequent infertility diagnoses in either men or women. This study represents the first large epidemiologic investigation of infertility after the smallpox vaccine. PMID- 18382141 TI - Polio: the Indian Sub-Continent's loosing war. AB - The recent debate about 'effective control' rather than 'complete eradication' for polio has put the whole Global Polio Initiative into question. The implications for this are indeed daunting, more so for the Indian Sub-Continent, which hosts a huge chunk of the global polio load every year. The tremendous success achieved in combating polio during the previous decade in India and Pakistan is reason enough to continue on with the vaccination efforts. However, other subsidiary issues including social stigma and a weak infra-structure have also to be put into the eradication equation to meet the goal. The Sub-Continent stands at the cross-roads, and time is ripe now to formulate a multi-faceted strategy to eliminate this crippling disease from this part of the world, and consequently give a much-needed boast to global eradication efforts. PMID- 18382143 TI - Stabilization of measles virus for vaccine formulation. AB - An attenuated live measles virus (MV) was characterized by several biophysical methods as a function of temperature and pH. Following a method developed previously, the resultant light scattering and spectroscopic data were synthesized into an empirical phase diagram that visually and simultaneously represents the entire data set. Using this empirically-based phase diagram, screening assays were developed to identify potential vaccine stabilizers. Various compounds are shown by these assays to inhibit the temperature-induced aggregation of viral particles, and also to protect the integrity of the viral envelope. Accelerated stability assays show that, upon thermal challenge, MV formulated with these excipients retains its infectivity to a significant extent. Thus, the enhanced physical stability produced by this method is shown to protect the biological activity of this important but labile vaccine. PMID- 18382142 TI - Immunogenicity and reactogenicity of acellular pertussis booster vaccines in children: standard pediatric versus a reduced-antigen content formulation. AB - Booster vaccination with a reduced-antigen-content dTpa, pediatric DTPa or adult Td vaccine in DTPa-primed children aged 4-6 years was evaluated. Immunogenicity and CMI was assessed one month and 3.5 years after vaccination. Symptoms were solicited for 15 days post-vaccination. There were no differences between groups in diphtheria or tetanus seroprotection or pertussis vaccine-response rates. Anti diphtheria and anti-PRN concentrations were higher after DTPa, but groups differences reduced over time. Non-significant trends toward reduced reactogenicity of dTpa were observed. Many factors influence vaccine choice at preschool age. The dTpa vaccine was as immunogenic and possibly better tolerated than DTPa at this age. PMID- 18382145 TI - Custom designing therapeutic cancer vaccines: delivery of immunostimulatory molecule adjuvants by protein transfer. AB - Attempts to create vaccines for humans against invading pathogens such as viruses and bacteria have met with tremendous success. The process of developing vaccines against these pathogens is greatly aided by the fact that they contain antigens that are entirely foreign to humans. Although the knowledge and strategies developed for designing vaccines against these microbes may be of use in developing cancer vaccines, the poor antigenicity and immunosuppressive ability of cancers pose major hurdles to vaccine development. Established tumors have not only withstood immune screening and selection pressure, making them poor stimulators of an immune response, but have also adapted mechanisms to continue evading immune surveillance by creating an immunosuppressive environment. Also, genetic differences in immune responses to an antigen among individuals result in an antigenic profile that varies from patient to patient. Cancers bear such great similarities to normal cells in the body that, on a molecular level, the differences between cancerous and non-cancerous cells are minor. Therefore, developing vaccines which use the host's own tumor tissues carries the risk of breaking tolerance to self-antigens that are present in the tumor tissue. Vaccination strategies that will optimally stimulate the immune system against tumor specific antigens under immunosuppressive conditions need to be developed. In practical terms, this calls for a method by which therapeutic vaccines may be custom-designed to treat cancers case by case. Ex vivo manipulation of dendritic cells and gene transfer of immunostimulatory molecules in ex vivo expanded tumors are being tested in both experimental models and also in human clinical trials. Some of them have met with limited success. Emerging technologies such as protein transfer, which make it possible to express immunostimulatory molecules on tumor cell membranes, offer the means to develop efficient tumor vaccines that are simple and fast, while being easy to store and administer in human patients. Progress in these techniques will move the cancer vaccine field a step closer towards realizing custom designed cancer vaccines in human clinical settings. PMID- 18382144 TI - An intranasal heat shock protein based vaccination strategy confers protection against mucosal challenge with herpes simplex virus. AB - Herpes simplex virus-1 (HSV-1) represents a significant obstacle for vaccine designers, despite decades of investigation. The virus primarily infects the host at vulnerable mucosal surfaces that progresses to lesion development, latency in nervous tissue, and possible reactivation. Therefore, protection at the site of infection is crucial. Mucosal adjuvants are critical for the development of an effective vaccine approach and heat-shock protein 70 (Hsp70) represents an attractive candidate for this purpose. This study demonstrates that Hsp70 coupled to gB498-505 from HSV-1 induced mucosal and systemic priming of CD8(+) T cells capable of protecting C57BL/6 mice against a lethal vaginal challenge. Elevated gB-specific cytotoxicity was observed in the spleen of mice immunized with conjugated Hsp70 and gB498-505. In addition, both vaginal IFNgamma levels and viral clearance were enhanced in mice mucosally immunized with Hsp70 and gB peptide versus peptide only control mice or mice receiving Hsp70 and a control peptide. These studies demonstrate that Hsp70 can be used as an effective mucosal adjuvant capable of generating a protective cell-mediated immune response against HSV-1. PMID- 18382146 TI - [Intra-abdominal infections: definitions and classification]. AB - Intraabdominal infections (IAIs) represent a wide variety of pathological conditions that involve lesions of all the intra-abdominal organs. They include both inflammation of single organs and any sort of peritonitis (primary, secondary, tertiary), where the severity of the disease often depends from the extension of the inflammation ((local or diffuse peritonitis). They include also the intra-peritoneal, retroperitoneal and parenchymal abscesses. The aim of current review is that of analyse the current definitions and classifications of intraabdominal infections. PMID- 18382147 TI - [Etiology, epidemiology and microbiological diagnosis of intra-abdominal infections]. AB - Intra-abdominal infections (IAIs) are commonly encountered in clinical practice. The etiology of these infections, often polymicrobial in nature, can be variable and usually includes organisms derived from the gut microbiota. In community acquired IAIs enterobacteria predominate (mostly Escherichia coli) in combination with anaerobes (mostly Bacteroides fragilis). In nosocomial IAIs, which can complicate abdominal surgery, other pathogens can also play a role, such as Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus spp. e Candida spp. Diagnostic microbiology of IAIs is complex and plays a relevant role especially in some situations (e. g. presence of foreign bodies, potential presence of resistant or uncommon pathogens, nosocomial infections in subjects with risk factors). Antibiotic resistance issues are currently encountered in most pathogenic species causing IAIs. Resistance affects all major classes of antimicrobial agents, often involving multiple classes and resulting in complex resistance phenotypes for which only a very limited number of drugs remain active. PMID- 18382148 TI - [Diagnosis of intra-abdominal infections: clinical findings and imaging]. AB - Abdominal sepsis carries a high morbidity and mortality. Intra-abdominal infectious complications are one of the most common infectious etiologies seen in critically ill patients. Approximately 30% of patients admitted to an ICU with intra-abdominal infection succumb to their illness, and when peritonitis arises as a complication of a previous surgical procedure, or recurs during ICU admission, mortality rates exceed 50%. Thus early detection and treatment is essential to minimize patient complications. Critically ill patients are often clinically non valuable due to distracting injuries, respiratory failure, obtundation, or other pathology. Even when patients can be examined, the clinical exam is frequently unreliable and/or misleading. The diagnostic approach to identify abdominal problems will differ depending upon the hemo-dynamic stability of the patient. Patients who have low systolic blood pressures, who are pressor dependent, may be too unstable to undergo studies that require trips away from the ICU or emergency department. Intra-abdominal pathology may be detected by ultrasound or diagnostic peritoneal lavage (DPL). When critically ill patients are stable enough to undergo some diagnostic evaluation of their abdomen the approach is somewhat simpler. Overall, computerized tomography (CT) is the imaging modality of choice for most intra-abdominal processes. For diagnosis of intra-abdominal conditions using CT scanning it is optimal if patients receive both oral and intravenous contrast. An exception to the use of CT scanning is evaluation of suspected biliary pathology, which is best imaged by ultrasound. It will identify cholecystitis with or without calculus and may show changes in the gallbladder or common bile duct associated with biliary obstruction. PMID- 18382149 TI - [Pharmacological rationale for choice of antibiotics for intraabdominal infections]. AB - The pharmacodynamic and pharmacokinetic characteristics of antimicrobial agents are the two fundamental pharmacological components which provide a rationale for the choice of therapy for intraabdominal infections, and especially serious infections. The most important PK-PD parameters are well known which can potentiate therapeutic efficacy. Antimicrobial agents can be subdivided into categories based on whether their activity is dependent on concentration or exposure time. Therefore, a correct dosing regimen for the time-dependent molecules (i.e. beta-lactams, linezolid, tigecycline) should prolong the maximum exposure time to maintain serum levels over the minimum inhibitory concentration (MIC). The concentration-dependent molecules, on the other hand, which include aminoglycosides and fluoroquinolones, should be given in order to reach maximum concentrations, since they are bactericidal in direct proportion to their concentrations and possess a prolonged post-antibiotic effect. PMID- 18382150 TI - [Current guidelines for the treatment of intra-abdominal infections]. AB - Intra-abdominal infections (IAIs) are a frequent cause of mobility and mortality. These infections can be caused by a wide variety of microorganisms, including both aerobes and anaerobes, and are often polymicrobial. Several studies observed that mortality depends on initiating early appropriate antimicrobial therapy. Moreover, the inappropriate choice of initial antibiotic therapy result in a longer hospital stay and higher costs of hospitalization compared with appropriate initial antibiotic therapy. The aim of current review is that of summarize the recommendations of several guidelines on the management of IAIs. PMID- 18382151 TI - [New options for treatment of intraabdominal infections: tigecycline]. AB - Tigecycline is a new antimicrobial agent; it is the first in a new class of antibiotics, the glycylcyclines, with properties conferring the ability to overcome many common resistance mechanisms, thus allowing its use for many serious and life-threatening infections for which the use of other antibiotics is no longer appropriate. It has a wide antibacterial spectrum including most methicillin-resistant Staphylococci, vancomycin-resistant Enterococci, ESBL producing Gram negative bacteria, and other MDR Gram negative bacteria such as Acinetobacter, and Stenotrophomonas. It has good antibacterial activity also against anaerobes and atypical pathogens. Tigecycline is available only as parenteral formulation. It has a high volume of distribution (>10 l/kg), and long half-life (36 hrs). It has been approved in USA and Europe for the treatment of complicated skin and soft tissue infections and for the complicated community acquired intra-abdominal infections. Phase III studies for treatment of community acquired and nosocomial acquire pneumonia, and sepsis sustained by multi-drug resistant pathogens are ongoing. PMID- 18382152 TI - [Principles and application protocols in the treatment of intra-abdominal infections]. AB - Antimicrobial therapy of intra-abdominal infection should consider its aetiology, which is generally polymicrobial, and its location with the aim of selecting the most suitable antimicrobial agents not only according to its spectrum but also to its pharmacokinetics profile Currently, both monotherapy and association therapy can be used, thanks the availability of newer drugs characterized by a wide range of antimicrobial activity both against aerobes and anaerobes. Antibiotic choice and duration therapy vary also according to the infection severity . PMID- 18382153 TI - [Hospital management of patients with complicated intra-abdominal infections: pharmacoeconomic evaluations]. AB - Due to their high incidence and large resource consumption, complicated intra abdominal infections (cIAIs) represent a heavy burden for the Italian National Health System (NHS) and the Italian society, with estimated annual costs of 1,5 and 3 billions Euro, respectively. The different strategies, monotherapy or antibiotic combinations, indicated for treating cIAIs induce significantly different acquisition and administration costs but substantially equivalent therapeutic results, with average clinical effectiveness rates of 70-80%. This apparent equivalence among different antibiotic protocols presumably depends on the widespread trend to individualize the therapeutic strategy according to the clinical severity and the community or nosocomial origin of cIAIs, as well as on some degree of non-appropriateness when empirically choosing a first-line antibiotic. The average cost for the nosocomial management of cIAI patients depends on several factors: posology and antibiotic drug acquisition and administration costs, days of antibiotic therapy, mix of antibiotic schedules, rates of the therapeutic failures that induce further drug consumption, prolong hospitalization and often require re-intervention and ICU utilization. The introduction in the therapeutic arsenal of a new antibiotic like tigecycline leads to a mild increase of the average antibiotic acquisition and treatment costs per patient: this increase is proportional to the percentage of patients treated with the new antibiotic. According to a decisional model, implemented on international outcome data and Italian costs, the mean cost for first-line antibiotic acquisition and the mean cost for first- and second-line antibiotic treatment represent respectively only 2% and 8% of the mean overall hospitalization cost. The mean hospitalization cost estimated by the model is noticeably higher than the mean value of DRG tariffs presumably reimbursed by the Italian NHS to hospitals for cIAI-related hospitalizations. Greater overall efficiency levels in the nosocomial management of cIAI patients are achievable mainly through the reduction of non-appropriateness rates in first line antibiotic choices and better treatment individualization, possible if the physician is offered the choice of as many valid therapeutic options as possible, in order to guarantee the best cure chances to each patient. PMID- 18382154 TI - Patient safety culture improvement tool: development and guidelines for use. AB - The Patient Safety Culture Improvement Tool (PSCIT) was developed to assist healthcare organizations in identifying practical actions to improve their culture. This article describes the development process of the PSCIT and provides a guide to using the PSCIT. The tool is based on a safety culture maturity model, which describes five stages of cultural evolution, from pathological to generative. The PSCIT consists of nine elements that cover five patient safety culture dimensions, namely, leadership, risk analysis, workload management, sharing and learning and resource management. Each element describes the systems in place at each level of maturity, enabling organizations to identify their current level of maturity and actions to move to the next level. The PSCIT should be used with caution as there is currently a lack of reliability and validity data. PMID- 18382155 TI - An evaluation of patient safety leadership walkarounds. AB - Patient safety leadership walkarounds (PSLWA) have been identified as an effective tool to improve patient safety culture. At Hamilton Health Sciences, after one year of monthly PSLWA in all clinical and service programs, 1,351 patient safety issues were identified, of which 64-80% have been resolved or have active improvement work in progress. Five hundred staff were invited to complete a process evaluation regarding the effectiveness of the current process of PSLWA. A total of 341 surveys were returned (68%). The overall evaluation demonstrated satisfaction with the process of PSLWA; 93% of those surveyed reported that they felt comfortable openly and honestly discussing patient safety issues and had an enhanced awareness of patient safety. Five areas of opportunity for process improvement were identified: scheduling, scripts, feedback, reporting and resolving issues deferred for an organization approach. PSLWA have offered an effective way to engage leadership and staff in open discussions about patient safety and collaborative approaches for solutions suggesting an enhanced patient safety culture. PMID- 18382156 TI - Implementation of a safety framework in a rehabilitation hospital. AB - This patient safety initiative was implemented at the Toronto Rehabilitation Institute, a fully affiliated hospital of the University of Toronto that operates in-patient and outpatient facilities on five sites. A working group was created to engage the leaders and employees in defining and implementing our "ideal" safety culture. A subset of this group became the Research Team, mandated to do the "discovery work" with external groups and internal stakeholders to provide valuable input for designing the safety culture model. This involved identifying the key components required to support a safety culture, testing this model with findings from the academic literature and best in class organizations and identifying the who, what, when, where and how of each key component. Future activities will focus on the integration of safety into existing programs, initiatives and policies, seeking feedback from staff, patients and families, and evaluating the effectiveness of our intervention and the extent of the culture change. PMID- 18382157 TI - Developing a patient safety plan. AB - Many healthcare organizations are focused on the development of a strategic plan to enhance patient safety. The challenge is creating a plan that focuses on patient safety outcomes, integrating the multitude of internal and external drivers of patient safety, aligning improvement initiatives to create synergy and providing a framework for meaningful measurement of intermediate and long-term results while remaining consistent with an organizational mission, vision and strategic goals. This strategy-focused approach recognizes that patient safety initiatives completed in isolation will not provide consistent progress toward a goal, and that a balanced approach is required that includes the development and systematic execution of bundles of related initiatives. This article outlines the process used by Hamilton Health Sciences in adopting Kaplan and Norton's strategy map methodology underpinned by their balanced scorecard framework to create a comprehensive multi-year plan for patient safety that integrates best practice literature from patient safety, quality and organizational development. PMID- 18382158 TI - Broadening the patient safety agenda to include safety in long-term care. AB - The recent patient safety literature has included less of an emphasis on long term settings than on research in the acute care sector. Recognizing this knowledge gap in our understanding of safety in the long-term care sector, the Canadian Patient Safety Institute, Capital Health (Edmonton) and CapitalCare (Edmonton) have collaborated to create a research and action agenda for improving resident safety in Canadian long-term care settings. This collaboration resulted in the development of a background paper highlighting the current state of the science and 14 key-informant interviews with stakeholders across Canada. The background paper subsequently informed an invitational round-table discussion. Key findings from the key-informant interviews as well as implications for research are described in this article. PMID- 18382159 TI - Nursing education: a catalyst for the patient safety movement. AB - Creating a culture of safety in healthcare systems is a goal of leaders in the patient safety movement. Commitment of leadership to safety in the Saskatchewan Institute of Applied Science and Technology (SIAST) Nursing Division has resulted in the development of the Patient Safety Project Team (PSPT) and a steady shift in the culture of the organization toward a systems approach to patient safety. Graduates prepared with the competencies necessary to be diligent about their practice and skilled in determining the root causes of system error in healthcare will become leaders in shifting the healthcare culture to strengthen patient safety. The PSPT believes this cultural shift begins with the education system. It involves modifications to curricula content, facilitation of multidisciplinary processes, and inclusion of theory and practice that reflect critical inquiry into healthcare and nursing education systems to ensure patient safety. In this paper the practical approaches and initiatives of the PSPT are reviewed. The integration of Patient Safety Core Curriculum modules for competency development is described. The policy for reporting adverse events and near misses is outlined. In addition, the student-focused reporting tool, the results and the implications for teaching in the clinical setting are discussed. Processes used to engage faculty are also addressed. PMID- 18382160 TI - Using ISMP Canada's framework for failure mode and effects analysis: a tale of two FMEAs. AB - Patient safety concerns in healthcare are not new or unexpected, and one goal of all healthcare organizations is to provide the safest possible care for patients and their families. With that goal in mind, Annapolis Valley Health, a rural district health authority in Nova Scotia, identified the need to develop expertise in the use of failure mode and effects analysis (FMEA) as a tool to promote quality processes within the organization. Staff members were aware of the value of this type of analysis but also recognized that real learning would best be achieved through completing an FMEA of an existing process or situation, rather than through a simulation or staff training. Annapolis Valley Health identified two high-risk situations requiring attention: transcription of medication orders for in-patients and overcrowding in the emergency department. The Institute for Safe Medication Practices Canada provided training and support to two staff teams and visited the organization eight months later for an update on progress. This article chronicles the journey of Annapolis Valley Health to improve patient safety through the application of FMEA to two high-risk processes for one of its hospital sites. PMID- 18382161 TI - Development of canadian safety indicators for medication use. AB - Reports of preventable illness due to medication errors are widespread in Canada. However, quantifying the magnitude of the problem has been hampered by a lack of measurement tools. Canadian-specific indicators, or performance measures, of safe medication use do not exist. The objective of this study was to develop a set of Canadian consensus-based indicators for the safe use of medication for both in patient and outpatient settings. A panel of 20 national experts was established from a convenience sample of experts representing medicine, nursing, pharmacy, research and decision-makers in hospitals and community settings across Canada. After creating a list of potential indicators from the literature, the final consensus set was chosen by the panel using a Delphi survey process via e-mail. After three rounds, consensus was obtained on 20 medication-use safety indicators: seven indicators were related to systems of care, five to prescribing/ordering, three to monitoring/assessment, three to medication administration, one to preparation and dispensing and one to purchasing/inventory management. Seventeen of the indicators measure a process of care (in contrast to health outcome); at least 10 have applications outside the in-patient setting. The resulting 20 medication-use safety indicators are diverse in scope and should be applicable in a variety of practice settings. These indicators may provide clinicians and decision-makers with valuable tools to assess the safety of medication-use systems. PMID- 18382162 TI - Medication safety in the operating room: teaming up to improve patient safety. AB - A medication safety project for operating rooms (ORs) was initiated under the leadership of the Departments of Anesthesia and Nursing with a representative from the Canadian Anesthesiologists' Society and the Institute for Safe Medication Practices Canada. The aims of the collaborative project were twofold: (1) to identify areas of exposure to risk and make recommendations to enhance medication safety within the hospital and (2) to inform the development of a medication safety checklist specific to the OR setting. The strategies developed and implemented during this project were aimed at reducing the risk of injury induced by medications. Attempts were made to use feasible best practices and managerial support systems for defined areas - in this case, medication-use systems for the ORs and associated patient care areas. The learning from this project will also inform the development of a medication safety checklist for use by other hospitals and OR settings. PMID- 18382163 TI - Survey of nursing perceptions of medication administration practices, perceived sources of errors and reporting behaviours. AB - In January 2003, St. Mary's Hospital Center in Montreal, Quebec, established an interdisciplinary Committee on the Systematic Approach to Medication Error Control to review the whole process of medication administration within the hospital and to develop a systematic approach to medication error control. A cross-sectional survey on medication administration practices, perceived sources of errors and medication error reporting of nurses, adapted from a nursing practice survey and medication variance report (Sim and Joyner 2002), was conducted over a two-week period in February 2004. The results were analyzed by years of experience (greater or less than five years) and patient care unit of practice. The perceived source of error most often cited was transcription (processing), and the second most frequently cited source was the legibility of handwritten medication orders (prescribing). The results demonstrate no significant difference in medication safety practices or in perceptions of errors by years of experience. Nurses appear to adapt to the safety culture of the unit rather quickly, certainly within their first five years on the unit. Good medication error reporting behaviour was noted, with no differences between all comparative groups within both years of experience and unit of practice. Quality improvement initiatives to improve the safety of medication administration practices have included the development of a nursing medication administration handbook, the revision of policies and procedures related to medication administration safety, the standardization of solutions and limited variety of high-risk medication dosages and the reduction of handwritten reorders. The need for ongoing education and information sessions on policies and procedures specific to safe medication practices for all nurses, regardless of years of experience, was identified. PMID- 18382164 TI - Communication and relationship skills for rapid response teams at hamilton health sciences. AB - Rapid response teams (RRT) are an important safety strategy in the prevention of deaths in patients who are progressively failing outside of the intensive care unit. The goal is to intervene before a critical event occurs. Effective teamwork and communication skills are frequently cited as critical success factors in the implementation of these teams. However, there is very little literature that clearly provides an education strategy for the development of these skills. Training in simulation labs offers an opportunity to assess and build on current team skills; however, this approach does not address how to meet the gaps in team communication and relationship skill management. At Hamilton Health Sciences (HHS) a two-day program was developed in collaboration with the RRT Team Leads, Organizational Effectiveness and Patient Safety Leaders. Participants reflected on their conflict management styles and considered how their personality traits may contribute to team function. Communication and relationship theories were reviewed and applied in simulated sessions in the relative safety of off-site team sessions. The overwhelming positive response to this training has been demonstrated in the incredible success of these teams from the perspective of the satisfaction surveys of the care units that call the team, and in the multi phased team evaluation of their application to practice. These sessions offer a useful approach to the development of the soft skills required for successful RRT implementation. PMID- 18382165 TI - Effectiveness of an Adapted SBAR Communication Tool for a Rehabilitation Setting. AB - Effective communication and teamwork have been identified in the literature as key enablers of patient safety. The SBAR (Situation-Background-Assessment Recommendation) process has proven to be an effective communication tool in acute care settings to structure high-urgency communications, particularly between physicians and nurses; however, little is known of its effectiveness in other settings. This study evaluated the effectiveness of an adapted SBAR tool for both urgent and non-urgent situations within a rehabilitation setting. In phase 1 of this study, clinical staff, patient and family input was gathered in a focus group format to help guide, validate and refine adaptations to the SBAR tool. In phase 2, the adapted SBAR was implemented in one interprofessional team; clinical and support staff participated in educational workshops with experiential learning to enhance their proficiency in using the SBAR process. Key champions reinforced its use within the team. In phase 3, evaluation of the effectiveness of the adapted SBAR tool focused on three main areas: staff perceptions of team communication and patient safety culture (as measured by the Agency for Healthcare Research and Quality Hospital Survey on Patient Safety Culture), patient satisfaction (as determined using the Client Perspectives on Rehabilitation Services questionnaire) and safety reporting (including incident and near-miss reporting). Findings from this study suggest that staff found the use of the adapted SBAR tool helpful in both individual and team communications, which ultimately affected perceived changes in the safety culture of the study team. There was a positive but not significant impact on patient satisfaction, likely due to a ceiling effect. Improvements were also seen in safety reporting of incidents and near misses across the organization and within the study team. PMID- 18382166 TI - Communities of practice: creating opportunities to enhance quality of care and safe practices. AB - A Communities of Practice (CoPs) approach was used to enhance interprofessional practice in seven clinical sites across Alberta. Participating staff were free to decide the area of practice to focus on and the actions to be implemented. All practice changes implemented by the CoPs related to either improving communications (e.g., introduction of joint care meetings) or information transfer (e.g., streamlining of admission and discharge processes). The practice changes contributed to more effective communication of information and more effective transitions of patients between providers, hence potentially reducing errors. The present study demonstrates that CoPs can enhance interprofessional communication and patient safety in traditional care delivery units. In contrast to more structured safety initiatives, sites were able to choose their area of focus. This ensures buy-in and enhances sustainability, making CoPs an interesting option for patient safety initiatives. PMID- 18382167 TI - Giving back the pen: disclosure, apology and early compensation discussions after harm in the healthcare setting. AB - In her recently published book After Harm, Nancy Berlinger shares a story about Bishop Desmond Tutu as he comments on the importance of restitution or compensation after an event that has led to harm. Transparency and disclosure are very much on the healthcare agenda in Canada. The increased interest in training providers for difficult conversations and disclosure is a positive sign. Using honest disclosure and apology as important interventions, organizations are beginning to adopt a more open approach to the concept of rebuilding trust after a patient has been harmed. But there continues to be significant reluctance to take the next logical step to solidify the fiduciary relationship between provider and patient - the willingness to enter into early discussions about compensation, non-monetary and otherwise. The Winnipeg Regional Health Authority has developed, with the participation of the facility insurers, a process to identify those cases in which it would be appropriate not only to offer an apology of responsibility but also to initiate discussions around the questions of restitution and compensation. The article describes the steps that led to the development of a detailed process map for such cases and shares the algorithm that has been adopted. As well, the potential challenges associated with such an approach when there are multiple liability and insurance providers are discussed. PMID- 18382168 TI - "It's safe to ask": promoting patient safety through health literacy. AB - The Manitoba Institute for Patient Safety launched "It's Safe to Ask" in January 2007. The communication and health literacy initiative is aimed at Manitoba's vulnerable populations and their primary care providers. Phase 1 includes a poster and brochure for patients and a toolkit for providers/organizations, pilot tested in six sites in Manitoba. Posters will serve as a symbol that dialogue is encouraged. Tools, available in 15 languages, provide patients and family members with three key questions to ask in healthcare interactions, tips on how to ask questions, and room for notes and listing of medications. The initiative will promote involvement in healthcare by patients, stronger communication between patient and provider, and reduction of risk for adverse events. "It's Safe to Ask" has been implemented in over 65 sites across Manitoba. A formal evaluation is underway. Phase 2 and 3 will enhance key tools and include interventions with specific populations. PMID- 18382169 TI - Canadian patient safety champions: collaborating on improving patient safety. AB - Patients for Patient Safety Canada champions have grown in numbers and purpose since their initiation into the World Health and Pan-American Health Organizations' Patients for Patient Safety initiative in May 2006. The 25 Canadian patients and family members not only share their adverse event experiences but are actively engaged in collaboration with health professionals, administrators and decision-makers to initiate proactive patient safety strategies. Their intention is to have their stories heard as tools for learning. They also wish to raise local, regional and national awareness of patient safety problems. The different patient and family stories and experiences share some common issues and suggested solutions that might make a difference in patient safety. One key solution is involving patients and families not only in discussions of treatment and follow-up when adverse events occur but also proactively on patient safety advisory committees. These actions would acknowledge a common interest in seeing that the right things are done. Patients and families share the common interest of all those advocating for patient safety, namely, First do no harm (attributed to Hippocrates, circa 470-360 B.C.). The patients and families of Patients for Patient Safety Canada are a group of committed, dedicated individuals who should be acknowledged for sharing their experiences and trying to make a difference in patient safety. PMID- 18382170 TI - Improving patient safety through a multi-faceted internal surveillance program. AB - Surveillance, a method used in epidemiology to study the incidence, distribution and control of disease, is an important means of gathering and analyzing information that can be used as needed to effect change. Surveillance has been an important component of the Blueprint for Patient Safety at the Hospital for Sick Children to identify potential and existing vulnerabilities and failures and put measures in place to avoid and mitigate any harm. Reviewing internal reports and actively seeking vulnerabilities has allowed us to make important changes to improve patient safety at the hospital. In this article, we review four internal surveillance strategies that have been particularly successful in driving change safety reports, morbidity and mortality reviews, patient safety walkarounds and shoe leather infection control rounds - and discuss the successes and challenges we have experienced. PMID- 18382171 TI - Safe medication swallowing in Dysphagia: a collaborative improvement project. AB - Episodes of choking during medication administration to patients with dysphagia prompted a chart audit and caregiver interview to identify system problems that allowed inappropriate drug administration to occur. Sixty elderly patients residing on two patient care areas in a 500-bed complex continuing care facility were studied. The audit explored the actual nursing medication administration methods and compared this to the information obtained from various communication tools including instructions that appeared on the medication administration record (MAR), the current diet order, the recommendations of the speech-language pathologist (SLP) and comments on the nursing care plan. The audit yielded a number of discrepancies between nursing actions and the instructions obtain from these sources. We proposed that changes to the process of communicating medication swallowing recommendations among team members would lead to greater patient safety. Major practice changes included the use of standardized language by the SLP when making recommendations, the writing of SLP recommendations in the doctor's orders, the inclusion of SLP recommendations on the MAR and the creation of a "dysphagia alert" on the pharmacy computer system. An educational intervention was conducted to implement process changes. Its effectiveness was evaluated using a pre- and post-test and a participant satisfaction survey. A post-implementation audit showed compliance with the practice change. In summary, process changes were implemented to improve compliance with SLP medication related swallowing recommendations and to prevent the inadvertent prescribing, dispensing or crushing of sustained-release medications in patients with dysphagia. PMID- 18382172 TI - An intervention program to reduce falls for adult in-patients following major lower limb amputation. AB - A qualitative and quantitative assessment was conducted regarding falls sustained by in-patients receiving rehabilitation therapy following major lower limb amputation at the Glenrose Rehabilitation Hospital. During the nine-month assessment period, 18 of 58 patients in the amputee unit experienced a fall, of which 17% resulted in a moderate injury. The majority of falls occurred during patients' use of a wheelchair (14 of 18) and involved poor balance (nine of 14). Patient wheelchair self-transfers accounted for 71% (10 of 14) of the falls, while sitting in the wheelchair and reaching represented 29/ (four of 14). The hospital's rehabilitation program teaches patient safety including using assistive devices such as wheelchairs but did not include a comprehensive graded learning path to monitor patients' ongoing risk for falls. Based upon the data collected, an intervention program was initiated to improve patient safety and reduce the number of falls. The multidisciplinary program encompassed aspects ranging from an environmental assessment of the patients' room to medication management, continuous patient wheelchair skills training and alteration of the care plan. The effectiveness of the intervention program was assessed through a series of interviews and questionnaires administered to medical personnel. This article presents the preliminary data collected during the first three months of the six-month study. Overall, satisfaction has significantly improved as a direct result of the intervention program. The article provides evidence-based interventions that improve safety for a subset of in-patients known to be susceptible to falls when using wheelchairs. Other in-patient groups will also benefit from these findings as many are universally applicable. PMID- 18382173 TI - Using human factors methods to evaluate the labelling of injectable drugs. AB - Adverse drug events, including in-hospital medication errors, are a well documented world-wide problem. This interdisciplinary team set out to examine the issues related to the labelling of injectable drugs. We sought answers to the following two questions: (1) To what extent do injectable drug labels adhere to existing Canadian design practice recommendations and regulations for labelling and (2) is there a need to make changes to the recommendations or regulations for labelling of injectable drugs in Canada? The project contained three phases. The first phase involved taking a sample of vials and ampoules from a hospital pharmacy and identifying adherence to the 1999 Canadian Standards Association standard for the labelling of drug ampoules, vials and prefilled syringes, as well as with the Canadian (Health Canada) Food and Drug Regulations for labelling. The second phase involved a failure mode and effects analysis of the label-reading process in order to identify information on the label considered critical for safe medication use. The third phase involved a preliminary human factors experiment addressing one problem identified with existing labels. Our finding is that existing injectable drug labels do not adhere sufficiently to available best design standards for labels and also do not adhere to all Canadian Food and Drug Regulations. Recommendations are made to inform future enhancements to labelling standards, guidelines and regulations. PMID- 18382174 TI - Prevention of ventilator-associated pneumonia in the calgary health region: a Canadian success story! AB - This article describes the experiences of a Canadian multidisciplinary critical care team striving to reduce the incidence of ventilator-associated pneumonia (VAP). Several interventions, including a VAP bundle, were used and applied across a health region. Our regional VAP rate has seen a steady decline over the past 12 months and has been largely under our goal of 9.8 cases per 1,000 ventilator-days. The team's success in lowering VAP has provided the momentum for sustained improvement, which has spread to other areas. PMID- 18382175 TI - An evaluation of a fall management program in a personal care home population. AB - Falls are a common problem among institutionalized adults, often resulting in serious negative consequences (Tideiksaar 2002). Fortunately, many of these falls are preventable (Tideiksaar 2002). However, there has been a recent shift from a fall "prevention" approach to one of fall "management," which aims at preventing injuries rather than falls. Falling is regarded as indicative of activity, which strengthens muscles, improves balance, and ultimately reduces the risk of falling (North Eastman Health Association Inc. 2005). For this research, the effectiveness of a fall "management" program that has been implemented in five provincial personal care homes "PCHs" in a Manitoba rural regional health authority will be evaluated. Fall-related administrative data will be analyzed to determine if there are differences (i) within the study sites over time (from pre to post-intervention) and (ii) between the study and comparison sites. Qualitative information from staff interviews and chart audits will supplement the quantitative information. PMID- 18382176 TI - Practice changes to improve delivery of surgical antibiotic prophylaxis. AB - Timely administration of appropriate antibiotics preoperatively can decrease the incidence of surgical site infection. We evaluated compliance with quality indicators in the delivery of antimicrobial surgical prophylaxis at The Ottawa Hospital and assessed the impact of a change to the hospital's Surgical Prophylaxis Policy. An audit in 2002 revealed improvement was necessary in the timing of preoperative doses, dosing for patients with a high body mass index, and intra-operative redosing. As a result, a multidisciplinary group was formed and a new surgical prophylaxis policy was approved. The policy included administration of preoperative doses by the anesthesiologist, and an automatic substitution for higher doses of antibiotics for select patients. This practice change resulted in significant improvements to the preoperative timing and dosing in subsequent audits. A mechanism to address intra-operative redosing will be implemented. PMID- 18382177 TI - Enteral levodopa/carbidopa infusion in advanced Parkinson disease: long-term exposure. AB - OBJECTIVES: In patients with advanced Parkinson disease, levodopa/carbidopa formulated as a gel suspension (Duodopa) permits continuous delivery into the small intestine using a portable pump, resulting in less variability in levodopa concentrations and fewer motor fluctuations and dyskinesias than with oral levodopa administration. This is a retrospective analysis of the long-term clinical experience with this agent. METHODS: All but 1 of the patients who had received enteral levodopa infusion treatment between January 1, 1991, and June 30, 2002, consented to a review of their hospital charts. RESULTS: Of the 65 patients with initial testing of the treatment, 86% opted for continued treatment via percutaneous endoscopic gastrostomy or gastrojejunostomy. Total exposure to levodopa infusion was 216 patient-years (mean, 3.7 years). Maximum treatment duration was 10.7 years. Fifty-two patients were treated for 1 year or longer. The adverse effect profile of levodopa/carbidopa infusion was similar to that observed with oral administration of levodopa. Seven deaths occurred, all considered unrelated to the treatment. Intestinal tube problems, including dislocation of the intestinal tube to the stomach, were the most common technical problem, occurring in 69% of the patients during the first year. The optimal daily dose of levodopa decreased by an average of 5% during follow-up. CONCLUSIONS: The safety of enteral infusion of levodopa/carbidopa formulated as a gel suspension was found acceptable. For most patients, the technical challenges posed by the enteral infusion system were offset by the improvement in motor fluctuations and dyskinesias offered by this technique. PMID- 18382178 TI - Common drugs in acute ischemic stroke and their influence on outcome: an observational study in 1013 stroke patients. AB - OBJECTIVE: Various drugs, when used during acute stroke, may affect stroke outcome. Some advise against the use of certain drugs that may be potentially harmful when used in acute stroke. However, for many of such drugs, the evidence is scarce. Therefore, we studied the use of various drugs at stroke onset as independent predictors of outcome at 3 months. METHODS: In an observational study of 1013 acute ischemic stroke patients, medication on admission was registered and divided in 10 categories. Functional outcome at 3 months was rated using the modified Rankin scale as functional independent (Rankin 0, 1, and 2) or as functional dependent or dead (Rankin 3, 4, and 5, or 6). Independent predictive values of medication use on outcome were analyzed using multivariate regression modeling. RESULTS: Vasodilator use was independently associated with less favorable functional outcome at 3 months (odds ratio [OR], 0.37 [95% confidence interval {CI}, 0.16-0.89]; P = 0.025) in lacunar stroke, as was the use of diuretics (OR, 0.43 [95% CI, 0.22-0.82]; P = 0.010) in atherothrombotic stroke. Calcium antagonists were independently associated with better outcome at 3 months (OR, 2.64 [95% CI, 1.04-6.69]; P = 0.042) in cardioembolic stroke. CONCLUSIONS: Our study does not yield strong evidence in favor of testing various commonly used drugs as potential neuroprotectives in acute stroke. PMID- 18382179 TI - Ischemic stroke related to anabolic abuse. AB - Anabolic-androgenic steroid (AAS) abuse increased in recent years, and it is associated with numerous adverse effects. Few reports on ischemic stroke related to anabolic steroid abuse have been published. We report a case of a 26-year-old male amateur athlete who suffered a posterior territory ischemic stroke. No abnormalities were found in angiography and echocardiography studies, neither in hemostatic profile. His only significant risk factor was nonmedical use of stanozolol, an anabolic steroid. Anabolic steroids are capable of increasing vascular tone, arterial tension, and platelet aggregation; therefore, they are prone to produce atherothrombotic phenomena. Because of young people's widespread use of anabolic steroids, physicians should be aware of this kind of complication. PMID- 18382180 TI - Evaluation of open-label topiramate as primary or adjunctive therapy in infantile spasms. AB - OBJECTIVE: A multicenter open-label clinical trial was conducted to evaluate the clinical usefulness of topiramate (TPM) as primary or adjunctive therapy for infantile spasms in the postmarketing period in China. METHODS: Thirty-four centers participated in the trial. Patients included in the study had 1 or more seizures per day before treatment. One hundred twenty (22.1%) very young patients with an age younger than 6 month and 64.2% of patients were younger than 1 year at start of treatment. All patients received a starting dose of 0.5 to 1 mg kg d TPM twice daily. The dosage was increased by 0.5 to 1 mg kg d every 5 to 7 days up to 3 to 5 mg kg d. The resulting range of the total TPM dosage was 25 to 200 mg d (3.57-20 mg kg d), with a median value of 73.9 mg d. Seizure outcomes were measured by intention-to-treat analysis. Patients were seen by a neurologist, and their data were evaluated at the day of inclusion and after 4, 8, 12, 16, and 20 weeks (from visit 1 to visit 5) of treatment. RESULTS: Five hundred forty-four patients entered the study. After 20 weeks of TPM treatment, 239 patients (43.9%) were seizure-free. A higher proportion of patients in the monotherapy group than in the add-on therapy group showed a seizure rate reduction. An increase in seizure frequency was observed in 8 patients (1.5%) during the 20-week treatment period. Nineteen patients were withdrawn before completing the study, and in 46 cases, some data of the structured data files and questionnaires were missing. No efficacy of TPM treatment was recorded in these cases. Adverse effects occurred in 211 patients (38.8%). Most frequent side effects were anorexia and somnolence. CONCLUSIONS: Topiramate proved to be an effective and safe monotherapy and add-on therapy in patients with infantile spasms younger than 1 year. PMID- 18382181 TI - Cutaneous drug eruptions by current antiepileptics: case reports and alternative treatment options. AB - Serious cutaneous drug eruptions due to antiepileptics have been defined for many drugs like carbamazepine, diphenylhydantoin, phenytoin and valproate. In recent years, adverse cutaneous reactions due to the current antiepileptic drugs have also been reported. In this paper, two cases are presented: a 48-year-old female receiving gabapentin for postherpetic neuralgia who developed leukocytoclastic vasculitis after 8 weeks and a 23-year-old male receiving lamotrigine for epileptic seizures who developed toxic epidermal necrolysis (TEN) in 15 days. Alternative therapy approaches with practical suggestions are also discussed. PMID- 18382182 TI - Topiramate in essential tremor: findings from double-blind, placebo-controlled, crossover trials. AB - OBJECTIVE: To evaluate topiramate in adults with essential tremor. METHODS: This report represents the combined results of 3 randomized, double-blind, placebo controlled, crossover trials that followed a common protocol. Study subjects were adults (> or =18 years old) who had untreated or treated moderate to severe essential tremor involving upper extremities. Patients were randomized to a double-blind sequence of topiramate (400 mg/d or maximum tolerated dose) then placebo (n = 30) or placebo then topiramate (n = 32). A 2-week washout period separated 10-week double-blind treatment phases. Upper extremity tremor was assessed using the Fahn-Tolosa-Marin tremor rating scale (TRS). The primary efficacy measure was the TRS total score at the final visit for patients providing on-treatment data in both double-blind treatment periods. Secondary efficacy measures included change from baseline in TRS total score and in TRS subscale scores for tremor severity, motor task performance, and functional disability. RESULTS: A total of 62 patients were enrolled. Total tremor score was significantly (P < 0.0001) lower with topiramate (28.7 +/- 1.0) vs placebo (37.0 +/- 1.0). The change from baseline in TRS total and subscale scores was significantly greater (P < or = 0.005) with topiramate treatment (mean score reduction, 7.7-11.8 vs 0.08-2.0). Of the 28 patients who discontinued without completing both treatment periods, adverse events accounted for 13 of 18 discontinuations during topiramate treatment and 5 of 10 discontinuations during placebo exposure. Adverse events reported by 2 or more patients discontinuing topiramate were nausea (n = 3), paresthesia (n = 3), and concentration/attention difficulty (n = 2). CONCLUSIONS: Topiramate was associated with overall tremor reduction and improvements in tremor severity, motor task performance, and functional disability in patients with moderate to severe essential tremor. PMID- 18382183 TI - Epigenetic therapeutic strategies for the treatment of neuropsychiatric disorders: ready for prime time? AB - Chromatin remodeling is recognized as a major regulator of gene expression that can be influenced by inhibition of epigenetic mechanisms that result in stable, heritable, covalent modifications of histone proteins and their associated DNA. Epigenetically regulated covalent modifications are implicated in the pathogenesis of some forms of cancer and stimulated clinical trials of compounds selected for their ability to arrest cell division and promote differentiation of malignantly transformed cells. Chromatin remodeling may also be considered as a therapeutic target in diverse neuropsychiatric disorders such as Huntington disease and other neurodegenerative disorders characterized by expression of mutant proteins with expanded tracts of polyglutamine repeats, schizophrenia, and major depression. Ideally, these strategies will be relatively selective because epigenetic abnormalities may be most pronounced in specific cell types, and tissues and transcriptional dysregulation due to pathological covalent modifications involve only a small percentage of all the expressed genes in the human genome. To date, beneficial effects of epigenetic therapeutic interventions such as administration of histone deacetylase inhibitors have been observed in transgenic mice expressing mutant human DNA constructs of proteins with expanded polyglutamine repeats and other rodent models of neuropsychiatric disorders. The epigenetic therapeutic strategy has much promise, and its development will foster collaboration and cross fertilization between molecular and cell biologists, oncologists, psychiatrists, and neurologists. PMID- 18382184 TI - Optimizing long-term therapy for Parkinson disease: options for treatment associated dyskinesia. AB - There is currently no satisfactory treatment for dyskinesia in patients with Parkinson disease because most antidyskinetic strategies have the effect of aggravating Parkinsonian symptoms, and most pharmacological strategies for reducing "off" periods have increased dyskinesia as a treatment complication. Therefore, physicians and patients often have to balance treatment of its effects on Parkinsonian symptoms and on dyskinesia. In patients with advanced Parkinson disease, it is often not possible to induce periods of good mobility without dyskinesia. PMID- 18382185 TI - Acute kidney injury. AB - Diagnosis and classification of acute pathology in the kidney are major clinical problems. Azotemia and oliguria represent not only disease but normal responses of the kidney to extracellular volume depletion or decreased renal blood flow. Changes in urine output and glomerular filtration rate are therefore neither necessary nor sufficient for the diagnosis of renal pathology. However, no simple alternative for the diagnosis currently exists. By examining both glomerular and tubular function, clinicians routinely make inferences not only on the presence of renal dysfunction but also on its cause. However, pure prerenal physiology is unusual in hospitalized patients, and its effects are not necessary benign. Sepsis, the most common condition associated with acute renal failure in the intensive care unit, may alter renal function without any characteristic changes in urine indices, and classification of these abnormalities as prerenal will undoubtedly lead to incorrect management decisions. The clinical syndrome known as acute tubular necrosis does not actually manifest the morphologic changes that the name implies. A precise biochemical definition of acute renal failure has never been proposed, and until recently, there has been no consensus on the diagnostic criteria or clinical definition. Depending on the definition used, acute renal failure has been reported to affect from 1% to 25% of intensive care unit patients and has led to mortality rates ranging from 15% to 60%. From this chaos, two principles emerged: first, the need for a standard definition and, second, the need to classify the severity of the syndrome rather than only consider its most severe form. The RIFLE criteria were developed to achieve these goals, and the term acute kidney injury has been proposed to encompass the entire spectrum of the syndrome, from minor changes in renal function to requirement for renal replacement therapy. Thus, acute kidney injury is not acute tubular necrosis, nor is it renal failure. Small changes in kidney function in hospitalized patients are important and are associated with significant changes in short-term and possibly long-term outcomes. The RIFLE criteria provide a uniform definition of acute kidney injury and have now been validated in numerous studies. PMID- 18382186 TI - Epidemiology of acute kidney injury: how big is the problem? AB - OBJECTIVE: Acute kidney injury (AKI) is a complication that occurs frequently in hospitalized patients. In this article, we provide an overview of the literature on the epidemiology of AKI in hospitalized patients. PATIENTS AND SETTING: The overview is restricted to hospitalized patients, and most emphasis is put on intensive care unit patients. MEASUREMENTS AND MAIN RESULTS: The population incidence of less severe AKI and AKI treated with renal replacement therapy is approximately 2,000-3,000 and 200-300 per million population per year, respectively. These numbers are comparable with the estimates for severe sepsis and acute lung injury. Approximately 4-5% of general intensive care unit patients will be treated with renal replacement therapy, and up to two thirds of intensive care unit patients will develop AKI defined by the RIFLE classification. The incidence of AKI is increasing. Intensive care unit patients with AKI have a longer length of stay and therefore generate greater costs. In addition, AKI is associated with increased mortality, even after correction for covariates. Increasing RIFLE class is associated with increasing risk of in-hospital death. Patients with AKI who are treated with renal replacement therapy still have a mortality rate of 50-60%. Of surviving patients, 5-20% remain dialysis dependent at hospital discharge. CONCLUSION: AKI has a high incidence, comparable with acute lung injury and severe sepsis, and is associated with higher hospital mortality. PMID- 18382187 TI - Conventional markers of kidney function. AB - Acute kidney injury remains a serious clinical problem for intensive care unit patients, and its incidence is rising. The detection and diagnosis of acute kidney injury in the intensive care unit currently require use of conventional markers of kidney function, specifically, serum creatinine and urea levels and, less frequently, other urinary tests. These conventional markers are familiar to clinicians and have long been used at the bedside. However, these markers are clearly not ideal, each has limitations, and none reflect real-time changes in glomerular filtration rate or a genuine acute injurious process to the kidney. More importantly, these conventional markers can contribute to delays in recognition of acute kidney injury and, hence, delays to appropriate supportive and therapeutic interventions. The early detection and diagnosis of acute kidney injury should be a clinical priority. A diagnostic test or panel of tests that are capable of evaluating aspects both of kidney function and acute injury are desperately needed in critical care nephrology. Cystatin C has been shown superior to conventional markers and may assume a greater role in intensive care unit patients for detecting both early changes in glomerular filtration rate and evidence of acute injury. Other newly characterized markers of kidney function or acute injury have the potential to revolutionized the field of critical care nephrology and greatly improve the supportive and therapeutic management of intensive care unit patients with acute kidney injury. PMID- 18382188 TI - New biomarkers of acute kidney injury. AB - Acute kidney injury (AKI) represents a major clinical problem, with rising incidence and high mortality rate. The lack of early biomarkers has resulted in a delay in initiating therapies. Fortunately, the tools of modern science have revealed promising novel biomarkers for AKI, with potentially high sensitivity and specificity. These include a plasma panel (neutrophil gelatinase-associated lipocalin and cystatin C) and a urine panel (neutrophil gelatinase-associated lipocalin, interleukin 18, and kidney injury molecule-1). Because they represent sequential biomarkers, it is likely that the AKI panels will be useful for timing the initial insult and assessing the duration of AKI (analogous to the cardiac panel for evaluating chest pain) and for predicting overall prognosis with respect to dialysis requirement and mortality. It is also likely that the AKI panels will help distinguish between the various types and pathogeneses of AKI. It will be important in future studies to validate the sensitivity and specificity of these biomarker panels in clinical samples from large cohorts and from multiple clinical situations. Such studies will be markedly facilitated by multidisciplinary participation of various specialties (intensivists, cardiologists, surgeons) in AKI clinical studies and by the availability of commercial tools for the reliable and reproducible measurement of biomarkers across different laboratories. PMID- 18382189 TI - Can we prevent acute kidney injury? AB - OBJECTIVE: To review the literature on prevention of acute kidney injury (AKI). DATA SOURCE: MEDLINE- and PubMed-based review of literature published from 1965 to 2007. CONCLUSIONS: AKI is very common among critically ill patients. Even mild forms of AKI have significant attributable mortality. Hence, it is imperative that every effort to prevent AKI be made in clinical practice. However, there are very few interventions that have been shown to consistently prevent AKI. Measures such as adequate hydration, maintenance of adequate circulating blood volume and mean arterial pressure, and avoidance of nephrotoxins are still the mainstay of prevention. Loop diuretics and "renal-dose" dopamine have been clearly shown not to prevent AKI and may, in fact, do harm. Among the remaining pharmacologic options, N-acetylcysteine has the strongest evidence in prevention of AKI. Fenoldopam and theophylline need further investigation before being used to prevent septic AKI and contrast nephropathy, respectively. The role of prophylactic dialysis in preventing contrast nephropathy needs to be investigated further. PMID- 18382190 TI - Oliguria, volume overload, and loop diuretics. AB - Acute kidney injury (AKI) is commonly and increasingly encountered in patients with critical illness. In the past, epidemiologic studies have consistently found that oliguria further increases the risk of death from AKI. Compared with patients outside the intensive care unit (ICU), critically ill patients are more likely to have volume overload as a result of impaired solute and water excretion. Recently, broad changes have occurred in ICU practice, such as early goal-directed therapy in sepsis, which may further compound volume overload in the ICU patient with oliguric AKI. Evidence has also emerged to suggest that a positive fluid accumulation in ICU patients can unfavorably affect outcome. Thus, the ICU patient with oliguric AKI presents a dilemma with limited therapeutic options. These would include optimization of systemic hemodynamics, added fluid therapy, administration of loop diuretics, or finally, the initiation of renal replacement therapy. Interestingly, recent survey data and observational studies indicate that a majority of intensivists use loop diuretics, specifically furosemide, at some point during the course of illness in patients with AKI. Paradoxically, loop diuretics have been found in several clinical studies of patients with AKI to be potentially detrimental or, at the least, lack effectiveness for improving clinical outcomes. This contradiction between clinical practice and available evidence would suggest there is equipoise and need for higher-quality evidence to better characterize the role of loop diuretics in ICU patients with AKI. PMID- 18382191 TI - Vasoactive drugs and acute kidney injury. AB - The use of norepinephrine, and probably vasopressor therapy in general, in intensive care patients with hypotensive vasodilatation despite fluid resuscitation and evidence of acute kidney injury remains the subject of much debate and controversy. Although there is concern about the use of these drugs, these concerns are unfounded. At this time, the experimental and human data strongly suggest that, in these patients, vasopressor therapy is safe and probably beneficial from a renal, and probably general, point of view. On the basis of currently available evidence, in hypotensive vasodilated patients with acute kidney injury, restoration of blood pressure within autoregulatory values should occur promptly with noradrenaline and be sustained until such vasodilatation dissipates. The additional role of other vasopressors in these situations remains unclear. The addition of vasopressin may be helpful in individual patients, but widespread use is not supported by evidence. Alpha-dose dopamine has no advantages over noradrenaline and is not as reliably effective in restoring blood pressure and urine output. Its widespread use cannot be supported in patients with vasodilatation and acute kidney injury. Other vasopressor drugs such as epinephrine and phenylephrine may be similar in efficacy to noradrenaline. However, experience and available data with their use is vastly less than with noradrenaline. Adrenaline, in addition, is associated with hyperglycemia, hyperlactatemia, acidosis, and hypokalemia. Terlipressin appears useful in patients with acute kidney injury secondary to hepatorenal syndrome. Whether it is superior to noradrenaline in this setting remains uncertain, and more studies are needed before recommendations can be made. PMID- 18382192 TI - Renal repair and recovery. AB - OBJECTIVE: To review the cellular and molecular mechanisms of renal repair and recovery after acute kidney injury (AKI). DATA SOURCE: The data were summarized from published research articles. RESULTS: In AKI, there is an acute inflammatory response, epithelial cell necrosis and apoptosis, and shedding of epithelial cells into the tubular lumen. Recent work demonstrates that repopulation of damaged renal tubules occurs primarily from proliferation of tubular epithelial cells and resident renal-specific stem cells, with some contribution of paracrine factors from bone marrow-derived mesenchymal stem cells. In addition, growth factors seem to play a critical role in the repair process in animal models of renal injury. However, attempts to use growth factors in the clinical setting to attenuate human AKI or accelerate renal repair have not yet been successful. The endothelium also plays a critical role in the pathogenesis of AKI. Lastly, in human studies, the effect of dialysis on renal recovery remains poorly understood. CONCLUSIONS: Experimental animal models of AKI demonstrate that renal recovery and repair involves proliferation of tubular epithelial cells and stem cell populations and the coordinated contribution of multiple growth factors. Future efforts to improve recovery from AKI and improve patient outcomes may include novel therapies based on manipulation of populations of stem cells and augmenting repopulation of renal tubules. PMID- 18382193 TI - Long-term outcomes after acute kidney injury. AB - Acute kidney dysfunction is a common problem in intensive care units. It is not only associated with increased morbidity and mortality but also with increased healthcare costs. Limited healthcare budgets have now raised the issue of how much therapy should be dedicated to these critically ill patients. A precondition for any further discussion on this topic is the question on the long-term outcome and quality of life of these patients. However, only limited data are available in this field. In this review, we will focus on the existing literature, considering not only acute renal failure patients requiring renal replacement therapy but also those patients with mild or moderate impaired renal function. The intention of this review is to show that acute kidney injury is an important but often underestimated disease and a disease that deserves major attention because it is associated with impaired short- and long-term outcome. We will demonstrate that acute kidney injury patients requiring dialysis have a reasonable long-term survival rate and good quality of life. There is no doubt that aggressive intensive care unit treatment is justified in these patients, irrespective of the health costs. PMID- 18382194 TI - Pathophysiology of septic acute kidney injury: what do we really know? AB - Septic acute kidney injury accounts for close to 50% of all cases of acute kidney injury in the intensive care unit and, in its various forms, affects between 15% and 20% of intensive care unit patients. However, there is little we really know about its pathophysiology. Although hemodynamic factors might play a role in the loss of glomerular filtration rate, they may not act through the induction of renal ischemia. Septic acute renal failure may, at least in patients with a hyperdynamic circulation, represent a unique form of acute renal failure: hyperemic acute renal failure. Measurements of renal blood flow in septic humans are now needed to resolve this pivotal pathophysiological question. Whatever may happen to renal blood flow during septic acute kidney injury in humans, the evidence available suggests that urinalysis fails to provide useful diagnostic or prognostic information in this setting. In addition, nonhemodynamic mechanisms of cell injury are likely to be at work. These mechanisms are likely due to a combination of immunologic, toxic, and inflammatory factors that may affect the microvasculature and the tubular cells. Among these mechanisms, apoptosis may turn out to be important. It is possible that, as evidence accumulates, the paradigms currently used to explain acute renal failure in sepsis will shift from ischemia and vasoconstriction to hyperemia and vasodilation and from acute tubular necrosis to acute tubular apoptosis or simply tubular cell dysfunction or exfoliation. If this were to happen, our therapeutic approaches would also be profoundly altered. PMID- 18382195 TI - Acute kidney injury with iodinated contrast. AB - Diagnostic and interventional radiographic procedures in critically ill patients commonly depend on iodinated contrast media and consequently pose the risk of contrast-induced acute kidney injury. This is an important complication that accounts for a significant number of cases of hospital-acquired renal failure, with adverse effects on prognosis and healthcare costs. The epidemiology and pathogenesis of contrast-induced acute kidney injury, baseline renal function measurement, risk assessment, identification of high-risk patients, contrast medium use, and preventive strategies will be discussed in this article. An algorithm is suggested for the risk stratification and management of contrast induced acute kidney injury as it relates to patients undergoing iodinated contrast exposure during critical illness. Contrast-induced acute kidney injury is likely to remain a significant challenge for intensivists in the future because the patient population is aging and chronic kidney disease and diabetes are becoming more common. PMID- 18382196 TI - Abdominal compartment syndrome. AB - Acute renal failure frequently occurs in the intensive care unit as a primary or secondary event in association with trauma, surgery, or comorbid medical disease. An increasingly common thread linking surgical and medical disease management is the abdominal compartment syndrome. In particular, the rise of early goal directed therapy for the initial resuscitation and management of severe sepsis and septic shock is associated with an increased frequency of secondary abdominal compartment syndrome. This paper will explore the pathophysiology underpinning the abdominal compartment syndrome and its contribution to acute kidney injury and acute renal failure with regard to intra-abdominal pressure dynamics, preload limitation, and afterload augmentation. Diagnostic modalities and therapeutic interventions will be addressed as a means of reducing the frequency of acute kidney injury and acute renal failure in the critically ill. PMID- 18382197 TI - An overview of drug-induced acute kidney injury. AB - The complex nature of critical illness often necessitates the use of multiple therapeutic agents, many of which may individually or in combination have the potential to cause renal injury. The use of nephrotoxic drugs has been implicated as a causative factor in up to 25% of all cases of severe acute renal failure in critically ill patients. Acute tubular necrosis is the most common form of renal injury from nephrotoxin exposure, although other types of renal failure may be seen. Given that this is a preventable cause of a potentially devastating complication, a comprehensive strategy should be used to avoid nephrotoxicity in critically ill patients including: accurate estimation of pre-existing renal function using serum creatinine-based glomerular filtration rates, avoidance of nephrotoxins if possible, ongoing monitoring of renal function, and immediate discontinuation of suspected nephrotoxins in the event of renal dysfunction. PMID- 18382198 TI - Indications and timing of renal replacement therapy in acute kidney injury. AB - The optimal timing for initiation of renal replacement therapy in patients with acute kidney injury remains uncertain. Conventionally accepted indications include volume overload, hyperkalemia, metabolic acidosis, overt uremia, and even progressive azotemia in the absence of specific symptoms; however, precise definitions for these indications are lacking. Data from recent observational trials have suggested that early initiation of renal replacement therapy may be associated with decreased mortality; however, the results of these studies are inconclusive. Existing data on timing of initiation of renal replacement therapy in acute kidney injury that guide current clinical practice are summarized and issues that need to be addressed in future clinical trials are discussed. PMID- 18382199 TI - Dose and efficiency of renal replacement therapy: continuous renal replacement therapy versus intermittent hemodialysis versus slow extended daily dialysis. AB - Acute kidney injury represents an independent risk of death in the intensive care unit and significantly contributes to in-hospital mortality. The only accepted treatment of severe acute kidney injury so far is renal replacement therapy, which is not a causative therapy but rather a life-support treatment. Renal replacement therapy can be performed by several different techniques: intermittent hemodialysis, slow extended daily dialysis, peritoneal dialysis, or continuous renal replacement therapy. There is controversy about which technique should be used, which dosage should be selected for each therapy, and whether the technique and/or the dose of renal replacement therapy may impact survival in critically ill patients. After a careful review of the recent literature, definitive conclusions cannot be drawn: Trials are in most cases underpowered and conducted over many years, in which significant changes in the practice of acute dialytic techniques have taken place. Other studies have described therapeutic modalities requiring a high level of specific expertise in the field and generally not easily reproducible in the routine practice. While practitioners are waiting for the ultimate trial to be published, we think it is worth reporting some broad concepts and few suggestions for renal replacement therapy prescription derived from current evidence and from the available experience. PMID- 18382200 TI - Choice of therapy and renal recovery. AB - OBJECTIVES: To describe the impact of choice of therapy on renal recovery. DESIGN: Literature review. MAIN RESULTS: Randomized controlled trials conducted have not shown a benefit of continuous renal replacement therapy in mortality over intermittent renal replacement therapy. However, renal recovery is another important outcome for patients with acute kidney injury and may be affected differently by intermittent renal replacement therapy and continuous renal replacement therapy. Because of its rapid changes in fluid status and plasma osmolality, intermittent renal replacement therapy induces a decrease in venous return and can induce intradialytic hypotension. Because of this effect, intermittent renal replacement therapy may cause renal ischemia and delay renal recovery. Observational studies, including two large epidemiologic studies, suggest that continuous renal replacement therapy may be able to reduce chronic dialysis dependence. On the other hand, randomized controlled trials conducted so far do not support an effect of continuous renal replacement therapy over intermittent renal replacement therapy in relation to renal recovery. However, all of these randomized studies have significant limitations including sample size, study design, and randomization. CONCLUSIONS: Although there is much suggestive evidence that continuous renal replacement therapy may increase the rate of renal recovery, such evidence is insufficient to fully elucidate the impact of choice of therapy on this outcome. Appropriately planned trials will be required to address this issue. PMID- 18382201 TI - The future of extracorporeal support. AB - Extracorporeal therapy has expanded significantly over the past few decades from solely artificial renal replacement therapy. In patients with multiple organ dysfunction syndrome, it becomes necessary to provide multiple organ support therapy. Technological advances have opened the door to a multifaceted intervention directed at supporting the function of multiple organs through the treatment of blood. Indications for "old" therapies such as hemofiltration and adsorption have been expanded, and using these therapies in combination further enhances blood detoxification capabilities. Furthermore, new devices are constantly in development. Nanotechnology allows us to refine membrane characteristics and design innovative monitoring/biofeedback devices. Miniaturization is leading down the path of wearable/implantable devices. With the incorporation of viable cells within medical devices, these instruments become capable not only of detoxification but synthetic functions as well, bringing us closer to the holy grail of complete replacement of organ function. This article provides a brief overview of current and future direction in extracorporeal support in the critical care setting. PMID- 18382203 TI - Left ventricular noncompaction and cardiomyopathy: cause, contributor, or epiphenomenon? AB - PURPOSE OF REVIEW: To discuss unresolved issues pertaining to aetiology and diagnosis of isolated left ventricular noncompaction. RECENT FINDINGS: Left ventricular noncompaction may be sporadic or familial and is linked to mutations in mitochondrial, cytoskeletal, Z-line, and sarcomeric proteins. Severe childhood manifestations include fetal hydrops or sudden infant death syndrome. Adults with severe phenotypes have a similarly guarded prognosis due to heart failure, arrhythmia and thromboembolism. Conversely, healthy individuals may fulfil current imaging criteria for diagnosis. Left ventricular noncompaction is also observed in families with hypertrophic or dilated cardiomyopathy, casting doubt on its acceptance as a distinct disease entity. SUMMARY: The extent of myocardial compaction may be a continuous trait within the population. Sensitive imaging techniques may detect subtle variations in morphology that fall within the normal range, underscoring the need for more restrictive diagnostic criteria, as in mitral valve prolapse. Conversely, rather than being a root cause of myocardial dysfunction, left ventricular noncompaction may represent a secondary consequence of a genetic alteration, well-tolerated when the heart is otherwise normal. In the presence of a pathogenic mutation, disruption to myocyte function at a molecular level may be the primary disease determinant, with noncompaction arising as a maladaptive remodelling response that compounds the disease process through subendocardial ischaemia and fibrosis. PMID- 18382204 TI - Atrial fibrillation: evidence for genetically determined disease. AB - PURPOSE OF REVIEW: Atrial fibrillation is traditionally regarded as a sporadic, nongenetic disorder. Nevertheless, recent growing evidence points to an important heritable basis for atrial fibrillation, with significant genetic determinants. This paper reviews recent progress in understanding the role of genetic contributors to the pathogenesis of atrial fibrillation and its familial susceptibility. RECENT FINDINGS: Population-based studies have demonstrated a significant heritable component in atrial fibrillation, with specific contributors including single-gene mutations and single-nucleotide polymorphisms. Variants in both ion-channel and nonion-channel genes have been identified as potential atrial fibrillation-risk determinants. In addition, studies have pointed to interesting combined roles of genetic and environmental factors in atrial fibrillation pathogenesis, providing insights into gene-environment interactions. Clinical studies suggest that individual genetic profiles may determine the therapeutic response of atrial fibrillation. SUMMARY: Rapidly evolving work indicates that there are important genetic determinants of atrial fibrillation, and suggests that understanding these determinants will help us both to appreciate better the underlying pathophysiology and to provide new approaches in diagnosis, prevention and treatment of this common cardiac condition. PMID- 18382205 TI - Ion channel diseases in children: manifestations and management. AB - PURPOSE OF REVIEW: The identification of the first genes for the long QT syndrome (LQTS) in 1995 has spurred awareness among clinical cardiologists of cardiac ion channel diseases, resulting in more frequent diagnoses. Hence, there is a growing need to know more about the management of these complex and often lethal diseases. The present review addresses these practical issues. RECENT FINDINGS: Besides information on the genetic bases and prevalence, this review focuses primarily on the current state of therapy and management. This ranges from traditional and valid therapies to newer approaches still requiring validation to recent advances in gene-specific management. The significant problems generated by the growing use of the implantable cardioverter defibrillator (ICD) in children with the attendant impact on their quality of life are considered and attention is given to effective therapies which lessen the need to resort to an often life-saving therapy which in children, however, is burdened by major negative side-effects. SUMMARY: The findings described will allow practicing cardiologists to face their young patients affected by cardiac ion channel diseases with greater confidence to reach the best treatment and to have a better understanding of the special medical problems posed by these intriguing disorders. PMID- 18382206 TI - Clinical and molecular genetics of the short QT syndrome. AB - PURPOSE OF REVIEW: Sudden cardiac death in patients without structural heart disease remains a challenge in diagnostics and risk stratification. Genetically determined arrhythmias are a potential cause for a primary electrical disease. A recently discovered primary electrical disease is discussed. RECENT FINDINGS: The inherited short QT syndrome is a recently recognized genetic condition, which is associated with atrial fibrillation, syncope and/or sudden cardiac death. Attention has been focused on diagnostic ECG features, the identification of underlying mutations and mechanisms of arrhythmogenesis. SUMMARY: The short QT syndrome is clinically associated with atrial fibrillation, syncope and sudden cardiac death. A shortened QT interval (QTc <360 ms) and reduced ventricular refractory period together with an increased dispersion of repolarization constitute the potential substrate for reentry and life-threatening ventricular tachyarrhythmia. To date, gain-of-function mutations in KCNH2, KCNQ1, KCNJ2, encoding potassium channels and loss-of-function mutations in CACNA1C and CACNB2b, encoding L-type calcium channel subunits have been identified. The therapy of choice is the implantable cardioverter defibrillator in symptomatic patients. Quinidine has been shown to prolong the QT interval and to normalize the effective refractory periods of the atrium and ventricle in patients with short QT-1 syndrome. PMID- 18382208 TI - Left ventricular assist device-induced molecular changes in the failing myocardium. AB - PURPOSE OF REVIEW: There is considerable increase in the use of left ventricular assist devices for the treatment of severe heart failure. Traditionally viewed as a bridge to transplantation and more recently as a destination therapy, left ventricular assist device support is now recognized to offer potential for myocardial recovery through reverse remodeling, a potential that is further enhanced by combination with pharmacologic therapy. In this study, we examine the molecular changes associated with left ventricular assist device support and how these may contribute to the recovery process. RECENT FINDINGS: Studies in both patients and experimental models have demonstrated that improved function is associated with alterations in several key pathways including cell survival, cytokine signaling, calcium handling, adrenergic receptor signaling, cytoskeletal and contractile proteins, energy metabolism, extracellular matrix, and endothelial and microvascular functions. Moreover, the unique research opportunities offered by left ventricular assist device analysis are beginning to distinguish changes associated with recovery from those of mechanical unloading alone and identify potential predictors and novel therapeutic targets capable of enhancing myocardial repair. SUMMARY: Significant progress has been made toward revealing molecular changes associated with myocardial recovery from heart failure. These studies also offer new insight into the pathogenesis of heart failure and point to novel therapeutic strategies. PMID- 18382207 TI - Genetic determinants of cardiac hypertrophy. AB - PURPOSE OF REVIEW: Cardiac hypertrophy is a common phenotypic response of the heart to stimulants. It is associated with increased morbidity and mortality in various cardiovascular disorders. Genetic factors are important determinants of phenotypic expression of cardiac hypertrophy, whether in single-gene disorders or in complex traits. We focus on the molecular genetics of cardiac hypertrophy in various conditions with an emphasis on hypertrophic cardiomyopathy, a genetic paradigm of cardiac hypertrophic response. RECENT FINDINGS: The molecular genetic basis of cardiac hypertrophy in single-gene disorders has been partially elucidated. Likewise, the impact of genetics on the expression of cardiac hypertrophy in the general population has been demonstrated. Identification of mutations in the Z disk proteins has expanded the spectrum of causal mutations beyond the thin and thick filaments of the sarcomeres. In addition, modifier loci have been mapped and shown to impart considerable effects on the expression of cardiac hypertrophy in hypertrophic cardiomyopathy. Elucidation of the molecular genetics of sarcomeric hypertrophic cardiomyopathy and many of the phenocopies has highlighted the limitations of clinical diagnosis as a determinant of management and prognostic advice. The findings have raised the importance of diagnosis and treatment algorithms, which are based on both genotype and phenotype information. SUMMARY: Cardiac hypertrophy, regardless of the cause, is the phenotypic consequence of complex interactions between genetic and nongenetic factors. PMID- 18382210 TI - Heart failure 2008: an update for clinicians. PMID- 18382209 TI - Genetically determined myocarditis: clinical presentation and immunological characteristics. AB - PURPOSE OF REVIEW: Myocarditis is a clinically heterogeneous myocardial inflammatory disease, diagnosed by endomyocardial biopsy; it may be idiopathic, infectious, or autoimmune and may lead to dilated cardiomyopathy. Myocarditis and dilated cardiomyopathy represent different stages of an organ-specific autoimmune disease in genetically predisposed individuals. RECENT FINDINGS: In animal models, cell-mediated or antibody-mediated autoimmune myocarditis/dilated cardiomyopathy can be induced by viral infection or immunization with heart specific autoantigens, or can develop spontaneously in genetically predisposed strains. Susceptibility is based on multiple major histocompatibility complex and nonmajor histocompatibility complex genes. In patients the diagnosis of autoimmune myocarditis/dilated cardiomyopathy requires exclusion of viral genome on endomyocardial biopsy and detection of serum heart-reactive autoantibodies. They are directed against multiple antigens that are found in patients and relatives from about 60% of familial and nonfamilial pedigrees. They predict dilated cardiomyopathy development among relatives, years before disease. Some antibodies have functional effects on cardiac myocytes in vitro, in animals and possibly in a dilated cardiomyopathy subset, responsive to extracorporeal immunoadsorption. SUMMARY: In myocarditis/dilated cardiomyopathy, cardiac specific and disease-specific antibodies of IgG class are potential biomarkers for identifying 'at risk' relatives as well as those patients in whom, in the absence of active infection of the myocardium, immunosuppression, and/or immunomodulation may be beneficial. Future studies should better define the genetic basis of human autoimmune myocarditis/dilated cardiomyopathy. PMID- 18382211 TI - What do we know about anticoagulation in patients with heart failure? AB - PURPOSE OF REVIEW: The risk of thromboembolic events in heart failure patients is estimated to be in the range of 1-4.5% per year. To date, there are insufficient data to guide us in appropriate use of antithrombotic or antiplatelet drug therapy to decrease the risk of events in this population. This review will outline the existing literature on anticoagulation and heart failure. RECENT FINDINGS: Until recently, no randomized controlled data existed to identify the risks and benefits of anticoagulation in heart failure patients. Three recent trials have attempted to shed light on this topic. Unfortunately, these studies have been of limited value due to poor recruitment and are underpowered to definitively answer these questions. SUMMARY: Until additional randomized control data are available, the routine use of anticoagulation for heart failure patients cannot be advocated. Limited data suggest that the benefits of anticoagulation may outweigh the risks in the following categories: ejection fraction less than 20%; left ventricular systolic dysfunction and history of previous stroke; and known thrombus in the left or right ventricle. We eagerly await the completion of the Warfarin Aspirin Reduced Cardiac Ejection Fraction Study. PMID- 18382212 TI - Advances in the treatment of heart failure with a preserved ejection fraction. AB - PURPOSE OF REVIEW: Heart failure with preserved ejection fraction (HF-PEF) occurs in approximately 50% of patients with heart failure (HF) and is associated with high morbidity and mortality. A recent study demonstrated that, although survival improved significantly over time among HF patients with reduced ejection fraction (EF), there was no such trend toward improvement among patients with HF-PEF. Therefore, there exists an urgent need to develop effective treatment strategies specifically for patients with HF-PEF. Recently completed and ongoing research in the treatment of HF-PEF is reviewed in this article. RECENT FINDINGS: The two large randomized clinical trials completed in HF-PEF patients did not achieve statistical significance in benefit of renin-angiotensin system blockade on their primary combined endpoints of morbidity and mortality. Both trials, however, suggested the benefit of the angiotensin receptor and angiotensin-converting enzyme blockade on HF hospitalization. In addition, no clear benefit of beta blockers has been demonstrated specifically in patients with HF-PEF. SUMMARY: Current therapeutic recommendations for HF-PEF are aimed mostly at symptomatic management and treatment of concomitant comorbidities. Results of ongoing clinical trials further evaluating inhibition of the angiotensin and the aldosterone receptors as well as examining other novel therapeutic targets in HF PEF are keenly awaited. PMID- 18382213 TI - Nonischemic heart failure in diabetes mellitus. AB - PURPOSE OF REVIEW: Diabetic patients with heart failure have a poor prognosis. Although it has been demonstrated in animal models that metabolic maladaptation plays a pivotal role in contractile dysfunction of the heart, the understanding of 'diabetic cardiomyopathy' and its treatment in humans remains incomplete. RECENT FINDINGS: Epidemiological studies show that structural changes in the left ventricle can be demonstrated before onset of clinical diabetes. Diastolic dysfunction is the earliest manifestation that is associated with increasing level of serum-free fatty acids and worsening glycemic control. Spectroscopic and histologic evidence in the human myocardium indicates a maladaptive metabolic response in diabetes, characterized by intramyocellular triglyceride accumulation. Studies also suggest a link between myocardial isoform switching, calcium homeostasis and altered metabolism in the development of heart failure. However, treatment directed at deranged metabolic control in diabetes is effective only in animals, and not in humans. SUMMARY: Although clinical studies suggest the existence of 'diabetic cardiomyopathy', it is still difficult to prove causality. However, animal models and human studies suggest that systemic metabolic derangements may lead to metabolic, functional and structural maladaptation of the heart. The exact mechanisms of heart failure in diabetes remain elusive. PMID- 18382215 TI - Evidence for activation of immune system in heart failure: is there a role for anti-inflammatory therapy? AB - PURPOSE OF REVIEW: The efficacy of already established heart failure therapeutics limits new developments; however, there are still fertile grounds for growth with a better understanding of often overlooked mechanisms of disease. We will review evidence on inflammation and components of the inflammatory process occurring in patients with chronic heart failure, review different therapeutic techniques such as intravenous immunoglobin, plasmapheresis and immuno-adsorption. RECENT FINDINGS: The use of highly specific anti-inflammatory therapeutic strategies in large randomized trials such as anti-TNF-alpha has failed to show clinical benefits; however, by taking a broad spectrum anti-inflammatory approach, strategies as in the ACCLAIM trial have been effective in improving the outcome in a select group of patients. SUMMARY: There is substantial evidence of immune modulation promoting the downregulation of inflammatory cytokines thus increasing several important and potentially therapeutic anti-inflammatory cytokines. It will be necessary to design large multicenter trials to overcome the expected improvements of current therapies, which may overshadow results, and to finally elucidate the therapeutic contributions of immune modulation in heart failure. PMID- 18382216 TI - Emerging role of pharmacogenomics in heart failure. AB - PURPOSE OF REVIEW: The promise of pharmacogenomics is that it will one day result in targeted heart failure therapy that maximizes individual benefit and diminishes risk. Recent reports from the Beta Blocker Evaluation Survival and African American Heart Failure clinical trials provide a roadmap of how this promise may soon be realized. This review will discuss recent investigations of pharmacogenomics in heart failure, and the challenge of converting genomic heterogeneity into a usable clinical tool. RECENT FINDINGS: Genomic analysis from randomized clinical trials has been increasingly utilized to investigate genetic variables that affect drug response. Analysis from the Beta Blocker Evaluation Survival Trial, demonstrates the impact of beta-blockers on survival was primarily in patients with the beta-1 adrenergic receptor Arg389Arg genotype. The efficacy of angiotensin-converting enzyme inhibitors and the combination of isosorbide dinitrate and hydralazine differs in black and white heart failure cohorts. Initial reports from the African American Heart Failure Trial demonstrate the impact of aldosterone synthase polymorphism on left ventricle remodeling, outcomes and the impact of isosorbide dinitrate and hydralazine. Investigations from the African American Heart Failure Trial will continue to focus on determining the genomic bases for observed racial differences in therapeutic efficacy. An era of polygenic analysis, aided by Genome Wide Association Studies, should soon be upon us. SUMMARY: Modern clinical trials will increasingly have a pharmacogenetic component to allow more efficient targeting of therapeutics. Investigators are beginning to delineate the genomic basis for differences in drug efficacy between black and white heart failure cohorts. Pharmacogenomics will have an increasing role in the treatment of heart failure patients. PMID- 18382214 TI - Advances in the surgical treatment of heart failure. AB - PURPOSE OF REVIEW: Despite advancements in medical therapy, morbidity and mortality remain high. Surgical treatment of heart failure has been the subject of renewed focus, with a particular emphasis on applying the principles of evidence-based medicine to the evaluation of surgical therapies. The purpose of this review is to discuss emerging surgical therapies in heart failure, in particular, mechanical cardiac support and mitral valve repair. RECENT FINDINGS: The most widely established surgical therapy for heart failure is cardiac transplantation, but its impact is limited due to the limited number of donors. The Surgical Treatment for Ischemic Heart Failure study, a landmark evaluation of the role of coronary artery bypass grafting and surgical ventricular restoration in patients with ischemic heart disease and heart failure, has recently completed enrollment. Improvements in device design and patient selection appear likely to continue to improve outcomes with mechanical cardiac support in patients who are not deemed transplant candidates (destination therapy). Surgical repair of secondary mitral regurgitation is undergoing evaluation in the soon to be launched Surgery vs. Medical Treatment Alone for Patients with Mitral Regurgitation and Nonischemic study. SUMMARY: A variety of surgical therapies for heart failure are currently undergoing evaluation in randomized controlled trials. Data from these landmark studies will guide the application of surgical therapy in heart failure for the foreseeable future. PMID- 18382217 TI - Bibliography. Current world literature. Molecular genetics. PMID- 18382219 TI - Drug and alcohol services in middle-income countries. AB - PURPOSE OF REVIEW: The present review summarizes current research in middle income countries on drug and alcohol services for prevention, screening, treatment, care and rehabilitation between June 2006 and December 2007. RECENT FINDINGS: There is a dearth of scientific literature on prevention, treatment and rehabilitation in middle-income countries and no novel effective approaches reported during the review period. The Lancet Series on Global Mental Health showed only 0.7% of all papers reviewed were on low-income and middle-income countries. Several studies in these countries confirmed the effectiveness of brief interventions in treating alcohol use disorders at a primary care level. One study on alcohol screening in Brazil produced a promising screening tool. Other studies reported the availability of pharmacological and nonpharmacological treatments for alcohol and drug use disorders. Overall, poor accessibility to services and delayed onset of treatment persist. Political and cultural environments play a crucial role in providing services to fulfill treatment needs. SUMMARY: During the review period, reports from middle-income countries on alcohol and drug services were very limited. A few studies in some countries confirm the effectiveness of brief interventions in primary care settings. There is an obvious need for more research on alcohol and drug treatment services in middle-income countries. PMID- 18382220 TI - Ecstasy and other club drugs: a review of recent epidemiologic studies. AB - PURPOSE OF REVIEW: This review highlights the epidemiologic research on club drug use in the past year, with a focus on clinical epidemiology, social epidemiology, new methodological approaches, and alternative explanations for drug use behaviors. RECENT FINDINGS: Although 3,4-methylenedioxy-N-methylamphetamine (MDMA) or ecstasy is currently classified as a type of hallucinogen and its withdrawal is not recognized in the Diagnostic and Statistical Manual of Mental Disorders, there is evidence for the association of withdrawal symptoms with MDMA abstinence. Findings from latent class analysis indicate that MDMA users have a significantly higher risk of dependence than lysergic acid diethylamide users. Research on sociodemographic factors associated with club drug use continues to be a main focus worldwide. New epidemiologic research methods have been developed to enable researchers to monitor real-time drug use behaviors and to conduct surveys on sensitive issues in public places. In addition to traditional behavioral models, researchers began to examine the club drug phenomenon in the context of economic environment. SUMMARY: Recent findings on MDMA use further question the current drug classification in the diagnostic systems. Despite the continuous growth in the club drug research literature, there is no study on the influence of genetic factors on club drug use. More research in this area is needed. PMID- 18382221 TI - Substance use, post-traumatic stress disorder and violence. AB - PURPOSE OF REVIEW: The present review focuses on the co-occurrence of substance use disorder and post-traumatic stress disorder, with special attention to measurement and the role of violence as a contributor to the comorbidity. RECENT FINDINGS: Symptoms of post-traumatic stress disorder in the presence or absence of a post-traumatic stress disorder diagnosis are comorbid with several substance use dependencies and with a range of severity of substance use. SUMMARY: Lack of consistency in terms of substance use classification and measurement of post traumatic stress disorder across studies continues to hinder comparisons of rates of comorbid substance use disorder and post-traumatic stress disorder. More attention to the role of violence as a contributor to the comorbidity and its impact on treatment outcomes is warranted. PMID- 18382222 TI - Epidemiology of inhalant use. AB - PURPOSE OF REVIEW: The aim of the present article is to review recent research on the prevalence and correlates of inhalant use. RECENT FINDINGS: During the review period more prevalence studies have been conducted in the developing world, adding information to the ongoing studies that are periodically undertaken in the more developed countries. These studies suggest that inhalant use is widespread among children and adolescents and is increasing among females in the developing and developed world. Not all surveys report inhalants as a separate group from other illegal substances; data by type of inhalants are even rarer, and few studies address abuse or dependence. New evidence suggests lower reliability rates for the diagnostic criteria of dependence as compared with other substances, suggesting the need for a review including the evidence of withdrawal. Studies conducted in the period identify vulnerable groups and suggest an increased risk of injecting drug use, HIV, suicidality and psychiatric disorders among inhalant users. SUMMARY: The extension of inhalant abuse and its adverse consequences argues for greater efforts to advance classification and to increase knowledge through research, including the evaluation of prevention and treatment models. PMID- 18382223 TI - Drug use and health among prison inmates. AB - PURPOSE OF REVIEW: Health problems related to substance use are investigated and monitored throughout the world by both international organizations and governments. It is well recognized that prison inmates are a vulnerable group of the population. Research on this group is minimal compared with the general population. The problem of drug use and health consequences among this subpopulation is significant and needs to be explored. RECENT FINDINGS: A higher prevalence of drug use is found among prison inmates than among the general population. The health status of prison inmates seems to be lower because of both the inmates' behavior and the prison environment. Various communicable diseases are commonly found such as hepatitis C virus, HIV, and syphilis. SUMMARY: The health of prison inmates tended to be worse than in the general population, particularly for those who use drugs. Based on the principle of human rights and equity of access, healthcare services provided to prison inmates should be equivalent to those provided to the general population. PMID- 18382224 TI - Opioid dependence and pregnancy. AB - PURPOSE OF REVIEW: The management of opioid dependence during pregnancy has received considerable attention over the past three decades. Recent peer-reviewed literature in the fields of pregnancy and opioid dependence and neonatal abstinence syndrome has been evaluated and discussed. RECENT FINDINGS: Pregnant opioid-dependent women must be carefully managed to minimize harm to the fetus; therefore, standardized care for maternal health is required. In a multidisciplinary care system opioid maintenance therapy is the recommended treatment approach during pregnancy. Equivalent attention must be given to the treatment of neonatal abstinence syndrome, which occurs in 55-94% of neonates after intrauterine opioid exposure with a 60% likelihood of requiring treatment; heterogeneous rating scales as well as heterogeneous treatment approaches are often responsible for extended hospital stays. SUMMARY: Interpretation of available literature is confounded by several methodological flaws. In general, there is still a lack of evidence-based study designs for pharmacological treatment of these patients as well as neonatal abstinence syndrome. PMID- 18382225 TI - Plasma biomarkers of Alzheimer's disease. AB - PURPOSE OF REVIEW: The importance of biomarkers of Alzheimer's disease is increasing. The present review aims to offer a general view of plasma biomarkers of Alzheimer's disease and to discuss their relevance and limitations. RECENT FINDINGS: The broad overlap in the plasma amyloid beta protein (Abeta) levels between patients with Alzheimer's disease and control individuals indicates that the plasma Abeta level cannot differentiate cases of sporadic Alzheimer's disease from control cases. Although the significance of Abeta for diagnosing Alzheimer's disease is controversial, high plasma concentrations of Abeta40 and low plasma concentrations of Abeta42 indicate an increased risk of dementia. SUMMARY: The usefulness of biomarkers in cerebrospinal fluid has been shown by numerous studies; this test is not commonly used, however, and blood biomarkers are therefore preferred. Increasing evidence shows that the plasma Abeta concentration may be a premorbid marker for the risk of Alzheimer's disease. It may be used for therapeutic monitoring, diagnosis of Abeta deposition in the brain, and also as a surrogate genetic marker to identify novel genetic determinants of Alzheimer's disease. A potential role of plasma Abeta concentration as a marker of incipient dementia warrants further investigation. PMID- 18382226 TI - White matter hyperintensities in mid-adult life. AB - PURPOSE OF REVIEW: White matter hyperintensities on T2-weighted magnetic resonance imaging are frequent incidental findings in the brains of elderly individuals. Recent studies have reported that they may also be common in middle aged individuals, and their systematic evaluation in younger populations is necessary. RECENT FINDINGS: Incidental white matter hyperintensities are common in brains of healthy individuals in their 60s and may be seen as early as the 30s and 40s. They are associated with subtle functional impairment and higher prevalence of neuropsychiatric disorders. While cerebrovascular risk factors such as hypertension, diabetes, high homocysteine, and so forth, are known risk factors for white matter hyperintensities, a significant proportion of the variance is unexplained. Genetic factors, alone or in interaction with environmental factors, appear to be important. There is a slight excess of white matter hyperintensities in women, the basis for which is not understood. Longitudinal studies show that those with baseline lesions have a greater progression over time. SUMMARY: New imaging techniques present an opportunity to examine white matter pathology in great detail in younger populations. Standardized methods to examine such pathology and its determinants will help inform strategies for their prevention, which is an important component of a healthy ageing agenda. PMID- 18382227 TI - The neuropsychiatry of conversion disorder. AB - PURPOSE OF REVIEW: During the past two decades there has been a relative surge of interest in conversion disorder, and a multitude of studies have emerged on the subject. With continuing developments in neuroscience--mainly brain imaging--new applications to conversion disorder are being reported every year. RECENT FINDINGS: Diagnosis continues to represent a challenge, although neurological signs are increasingly being developed and validated to assist with this. Psychiatric co-morbidity diagnosed according to operational criteria is common. Brain imaging studies have brought some clues to understanding the pathophysiology of conversion disorder. SUMMARY: Evidence-based medicine requires reliable diagnostic criteria, and attempts have recently been made to validate some of the well known neurological signs of conversion disorder. From a psychiatric point of view, there is a need for greater understanding of the aetiology and mechanisms underlying conversion disorder and its relationship to other psychiatric disorders. Although advances have been made both in diagnostic methods and in the groundwork for a neurobiological model, no clear rationale for treatment is yet available and further research is strongly needed. PMID- 18382228 TI - Mirror neurons and the understanding of behavioural symptoms in psychiatric disorders. AB - PURPOSE OF REVIEW: Recent findings show that we can understand other people's actions, intentions and emotions through a mirror mechanism as if we performed the same actions and felt the same intentions or emotions (embodied simulation). The present paper reviews experimental evidence that this mechanism may be broken in some psychiatric disorders. RECENT FINDINGS: A mirror neuron system has been described in both monkeys and humans that allows one to map an observed action on a correspondent motor representation in the observer's brain. This mechanism has been involved in many higher motor functions ranging from action understanding to imitation and intention coding. A mirror mechanism has also been invoked in empathy, through an embodied simulation. SUMMARY: A dysfunction of the mirror neuron system may be at the root of the inability to empathize in patients with autism and may play a role in some negative and positive symptoms found in patients with schizophrenia. This opens up new perspectives in the interpretation of psychotic symptoms and possibly in developing therapeutic strategies. PMID- 18382229 TI - Does traumatic brain injury predispose individuals to develop schizophrenia? AB - PURPOSE OF REVIEW: Psychosis following traumatic brain injury has been studied in both prospective and retrospective studies that have been frequently limited by unclear criteria for classifying psychiatric syndromes or case ascertainment bias regarding traumatic brain injury. Schizophrenia is a psychotic disorder commonly attributed to the interaction of genetic vulnerability and environmental events, yet its relationship with traumatic brain injury has not been systematically reviewed. RECENT FINDINGS: The literature is not conclusive and highlights the challenges to observational research in conditions in which case ascertainment is difficult. The evidence supports a risk-modifying effect of traumatic brain injury in individuals who are genetically at risk for schizophrenia, but is less supportive of traumatic brain injury as an independent risk factor for schizophrenia in individuals without such risk. SUMMARY: Research in psychotic disorders following traumatic brain injury must distinguish schizophrenia from other psychotic syndromes, and take familial risk factors into consideration. PMID- 18382230 TI - The contribution of failing adult hippocampal neurogenesis to psychiatric disorders. AB - PURPOSE OF REVIEW: Failing adult neurogenesis is increasingly considered a factor in the pathogenesis and course of psychiatric disorders. The level of evidence in favor of such hypotheses varies, but disturbed cellular plasticity in the hippocampus may be a common aspect of several neuropsychiatric diseases. RECENT FINDINGS: This review covers the literature from mid-2006 to the end of 2007. We discuss studies and theoretical papers dealing with the contribution of adult neurogenesis to dementias and neurodegeneration, major depression, schizophrenia, and alcohol and drug abuse. Of these disorders, most progress has recently been made with schizophrenia for which, in contrast to the other conditions, suggestive genetic evidence exists (e.g. Disc1, Npas3). SUMMARY: Failing adult hippocampal neurogenesis may not explain major depression, addiction or schizophrenia, but contributes to the hippocampal aspects of the disease. We propose that the key to a more thorough understanding of this contribution will come from increased knowledge on the functional relevance of new neurons in the hippocampus and better clinical data relating to symptoms possibly related to such function. Research on the molecular basis of adult hippocampal neurogenesis may help to explain how hippocampal aspects of these disorders develop. PMID- 18382231 TI - Brain reserve and the prevention of dementia. AB - PURPOSE OF REVIEW: To evaluate and synthesize recent evidence linking mental activity and dementia risk, which commonly invokes 'brain reserve' as the mediating construct. RECENT FINDINGS: Brain reserve has acquired several interpretations; however, the most reliable and practical definition focuses at the behavioural level by assessing frequency and range of participation in complex mental activities. Epidemiological research suggests a clear and consistent link of high brain reserve with reduced dementia risk. Furthermore, emerging clinical trials of cognitive exercise suggest that it may be effective for the prevention of longitudinal cognitive and functional decline. Recent animal studies implicate several mechanisms, including disease-dependent and disease-independent compensatory pathways. SUMMARY: Given the precipitous forecasts for dementia over the coming decades, effective preventive strategies are of utmost importance. Findings from brain reserve studies now meet many of the formal criteria for causal agency between complex mental activity and reduced dementia risk. Key clinical trials are therefore under way to test these claims and results are keenly awaited. PMID- 18382232 TI - Bibliography. Current world literature. Addictive disorder. PMID- 18382234 TI - Prostate cancer: targeting complexity. PMID- 18382235 TI - Current controversies in the treatment of high-risk prostate cancer. AB - PURPOSE OF REVIEW: Despite the well documented stage migration in prostate cancer, a substantial number of men still present to urologists with locally advanced or metastatic disease. RECENT FINDINGS: The beneficial role of prostatectomy has been affirmed in several studies examining its therapeutic impact in locally advanced, nonmetastatic prostate cancer. Adjuvant therapy with radiation or hormones appears to increase prostate-specific antigen relapse-free survival. Whether prostate-specific antigen relapse-free survival is an appropriate surrogate for overall survival remains unverified. The timing and duration of hormonal therapy continues to be debated. Hormone therapy administered 'too late' in the course of metastatic disease portends a shortened survival but possible side effects of androgen ablation must be considered. Several docetaxel-based combination chemotherapies for hormone refractory prostate cancer are being studied, but their efficacy in the neoadjuvant setting thus far has been limited. Progress in the palliation of bony metastases has resulted in a decrease in symptoms and skeletal events. SUMMARY: This review identifies seminal data that focus on controversial therapeutic dilemmas in prostate cancer. The literature of the last few years universally emphasizes the importance of a multidisciplinary collaboration in prostate cancer. It is only with this type of cooperation that essential research will continue and succeed. PMID- 18382236 TI - Focal therapy for prostate cancer. AB - PURPOSE OF REVIEW: Nowadays the treatment paradigm for localized prostate cancer is to distinguish patients with clinically relevant cancers who may benefit from radical treatment, or perhaps an organ-sparing approach, from the remainder who may not need intervention at the time of diagnosis. We review new concepts of parenchymal preservation as possible new frontiers in the treatment armamentarium for this malignancy. RECENT FINDINGS: For a select cohort of patients with low risk unifocal or unilateral prostate cancer lesions, a number of ablative treatment options for focal therapy are available with cryotherapy having the most clinical experience. Technologies that have the ability to be utilized for focal therapy include high-intensity focused ultrasound, brachytherapy, interstitial laser thermotherapy, stereotactic radio surgery, and vascular targeted photodynamic therapy. Further basic and animal research along with the conduction of large-scale randomized clinical trials demonstrating long-term disease-free survival and quality of life outcomes are necessary. SUMMARY: The concept of focal therapy is evolving with the understanding of the biologic variability (clinically aggressive, significant or insignificant) of various prostate cancer lesions that may require different treatment approaches. Minimally invasive, parenchyma-preserving therapies can assume a greater role in the treatment of unilateral or unifocal lesions, representing an alternative approach to the current treatment extremes of whole-gland treatment and watchful waiting. PMID- 18382237 TI - Current decision-making in prostate cancer therapy. AB - PURPOSE OF REVIEW: Prostate cancer continues to be the most prevalent cancer among American men. Localized prostate cancer is commonly diagnosed because of improved screening practices nationwide. Several options exist for the treatment of localized prostate cancer, and this review discusses the decision-making process facing patients diagnosed with this disease. RECENT FINDINGS: No one treatment for localized prostate cancer has proven superior to date. For this reason patients have been found to use a number of resources to make an informed decision. These include physicians, spouses, family, friends, and different media. Urologists serve as the primary and most influential physicians and play an important role in the decision-making process. Patients, however, are assuming a more active role in this process as time evolves, especially with ease of access to multiple information resources. SUMMARY: In deciding on a treatment for localized prostate cancer, patients must weigh the risks and benefits of each option. Urologists must provide patients with up-to-date information on these options and be aware of the different influences that surround these men during the decision-making process. PMID- 18382238 TI - Inventory of prostate cancer predictive tools. AB - PURPOSE OF REVIEW: We created an inventory of current predictive tools available for prostate cancer. This review may serve as an initial step toward a comprehensive reference guide for physicians to locate published nomograms that apply to the clinical decision in question. Using MEDLINE a literature search was performed on prostate cancer predictive tools from January 1966 to November 2007. We describe the patient populations to which they apply and the outcomes predicted, and record their individual characteristics. RECENT FINDINGS: The literature search generated 111 published prediction tools that may be applied to patients in various clinical stages of disease. Of the 111 prediction tools, only 69 had undergone validation. We present an inventory of models with input variables, prediction form, number of patients used to develop the prediction tools, the outcome being predicted, prediction tool-specific features, predictive accuracy, and whether validation was performed. SUMMARY: Decision rules, such as nomograms, provide evidence-based and at the same time individualized predictions of the outcome of interest. Such predictions have been repeatedly shown to be more accurate than those of clinicians, regardless of their level of expertise. Accurate risk estimates are also required for clinical trial design, to ensure homogeneous high-risk patient groups for whom new cancer therapeutics will be investigated. PMID- 18382239 TI - Assessing the quality of prostate cancer care. AB - PURPOSE OF REVIEW: Until recently, little was known about the quality of prostate cancer care in the United States. This article provides an overview of the methodology of quality of care research, reviews the available quality measures in prostate cancer and presents an overview of the existing literature on the quality of prostate cancer care in the US. RECENT FINDINGS: Researchers have applied methodologies developed in other conditions to construct tools to measure the quality of care in this disease. Initially, researchers from the RAND Corporation developed a preliminary performance measure set. This measure set was tested in a number of settings. Along with a number of clinical guidelines, the RAND measures served as the basis of new prostate cancer measures developed by the American Medical Association's Physician Consortium for Performance Improvement. Recent studies document that patients undergoing radical prostatectomy had worse documented compliance with quality indicators than those undergoing external beam radiotherapy. SUMMARY: There is clearly room for improvement in prostate cancer quality of care in the US. If providers do not take the initiative and address these shortcomings, providers and policymakers will implement changes that may not be in the best interests of patients. PMID- 18382241 TI - Prostate cancer biobanking. AB - PURPOSE OF REVIEW: This review is intended to provide an overview of 'best practices' pertaining to prostate biobanking. It addresses issues related to collection and appropriate annotation of prostate samples, ethics and administrative aspects of biorepository functioning. RECENT FINDINGS: Paraffin tissue microarrays have emerged as a significant mechanism for high throughput evaluation of markers of interest. In addition, modification of guidelines and definitions of 'human research' have served to provide mechanisms for expediting biological specimen disbursement. SUMMARY: A well developed biobank is a critical prerequisite for high-quality research. This review provides an outline of certain critical elements that would need careful attention as a prostate cancer biobank is developed. PMID- 18382240 TI - Clinical endpoints for drug development in prostate cancer. AB - PURPOSE OF REVIEW: Overall survival remains the benchmark in phase III settings of novel agents in castration-resistant metastatic prostate cancer. This review highlights many of the current potential early measures of response and clinical benefit that are worthy of future study and validation in this disease. RECENT FINDINGS: The clinical evaluation of novel agents in advanced prostate cancer remains challenging for several reasons. Men with metastatic prostate cancer often have bone-only disease in which formal radiologic response and progression criteria may not apply. Declines in serum prostate-specific antigen levels may be modest surrogates of response to cytotoxic agents such as docetaxel, but have not been validated for agents with novel mechanisms of action, such as antiangiogenic, immunologic, or cytostatic drugs. Novel radiologic imaging techniques such as PET scans are not yet validated for use in monitoring or staging advanced prostate cancer. Measures of delay, control, and palliation of metastatic disease such as pain response, time to progression and progression free survival, while appealing endpoints that may highlight the clinical benefit of novel agents, have been difficult to define rigorously and have not yet demonstrated adequate surrogacy for overall survival. SUMMARY: The measures of response highlighted in this review, if validated, may improve the current evaluation of novel agents in phase II settings and the potential accelerated approval of these agents. PMID- 18382242 TI - Inflammation, infection, and prostate cancer. AB - PURPOSE OF REVIEW: Recent evidence suggesting that inflammation and infection play a role in the etiology of prostate cancer, and the discovery of a novel virus in men with a genetic susceptibility to prostate cancer is reviewed. RECENT FINDINGS: Almost 20% of visceral cancers worldwide have proven infectious causes. There is substantial histologic, molecular genetic, and epidemiologic evidence that infections and inflammation are also important in the pathogenesis of prostate cancer. The R462Q allelic variant in RNASEL, an antiviral gene important in the innate immune response to viral infections, increases susceptibility to prostate cancer while resulting in vitro in deficient antiviral defenses, suggesting that prostate cancer could be caused by a virus. The study of R462Q carriers led to the discovery and biologic characterization of a novel retrovirus, xenotropic murine leukemia-related virus, isolated and cloned from prostate tissue of affected men. Biologic studies of this virus show that it is sensitive to inhibition by interferon and its downstream mediator, RNaseL, and that its DNA has integrated into the DNA of some men with prostate cancer. SUMMARY: Inflammation triggered by infection and other causes underlies the development of prostate cancer, and xenotropic murine leukemia-related virus is a candidate etiologic agent. Ongoing studies seek to define its oncogenic potential and pathogenesis. PMID- 18382243 TI - The critical use of population-based medical databases for prostate cancer research. AB - PURPOSE OF REVIEW: Population-based investigations comprise a significant portion of the prostate cancer literature and contribute considerably to our knowledge of this disease. It is important to recognize, however, the inherent limitations of observational study designs. In this review, we will highlight important population-based studies in prostate cancer of high methodological quality using the framework of the recently published Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) guidelines. RECENT FINDINGS: Recent findings of population-based studies of prostate cancer have generated key advances in our understanding of prostate cancer outcomes and epidemiology, which are highlighted in this review. The STROBE criteria, which were developed to promote the transparent reporting of observational studies, provide an excellent framework to determine the validity of conclusions drawn from large population based studies. SUMMARY: Population-based research is a powerful tool for understanding prostate cancer, but clinicians and investigators must interpret findings appropriately. PMID- 18382244 TI - Shifting focus to cancer survivorship in prostate cancer: challenges and opportunities. AB - PURPOSE OF REVIEW: As strides have been achieved in cancer control, issues faced by survivors have become increasingly relevant. Currently, there are more than 10 million cancer survivors living in the US and consequently cancer survivorship has become a major health priority. Approximately 2 million prostate cancer survivors face a host of long-term and late effects following cancer therapy. Although progress has been made in recognizing many of the issues faced by survivors, gaps in comprehensive surveillance and management still exist. RECENT FINDINGS: In its recent report, From Cancer Patient to Cancer Survivor: Lost in Transition, the Institute of Medicine outlined deficits in the care provided to cancer survivors and proposed several mechanisms to improve the coordination and quality of follow-up care. Measures to achieve these objectives include improving communication between healthcare providers through a survivorship care plan, developing an evidence base to better support surveillance guidelines, and assessing different models of survivorship care. SUMMARY: Potential shortcomings in the recognition and management of ongoing issues faced by prostate cancer survivors may impact the overall quality of long-term care in this growing population. Although programs to address these issues have been proposed, they are in the earliest phase of development. PMID- 18382245 TI - Review of recent evidence in support of a role for statins in the prevention of prostate cancer. AB - PURPOSE OF REVIEW: We examine the potential chemopreventive role statins may have in prostate cancer, highlight the basic science supporting this role and analyze the human data regarding the association between statin use and prostate cancer. RECENT FINDINGS: Basic scientific evidence suggests that, through cholesterol and noncholesterol-mediated mechanisms, statins inhibit many pathways of cancer formation and progression. A handful of observational studies found statin use was associated with reduced prostate cancer risk, though others found no association. In the last year, however, four large prospective studies have observed similar reductions in the risk of advanced prostate cancer with essentially no reduction in the risk of overall prostate cancer. This may, in part, explain why previous studies, including large metaanalyses of clinical trials of statins in the prevention of cardiovascular outcomes, did not observe any association between statin use and overall prostate cancer risk. SUMMARY: The exact association between statin medication use and prostate cancer, and whether this association is causal in nature, remains unclear. Recent evidence, however, is encouraging, particularly for reducing the risk of advanced disease. Thus, while at present there are insufficient data to recommend all men start taking a statin medication regardless of their cholesterol profile, the rationale to move forward with further research is clear. PMID- 18382246 TI - Bibliography. Current world literature. Prostate cancer. PMID- 18382249 TI - Progress in prevention: disseminating awareness of stroke symptoms: a call to action. PMID- 18382248 TI - Physical activity as a potential mechanism through which social support may reduce cardiovascular disease risk. AB - Social support has been associated with a reduced risk of cardiovascular disease (CVD). It has been suggested that the protective effect of social support is mediated through the autonomic nervous system and/or unhealthy lifestyle behaviors, but data are sparse, especially in diverse populations. The purpose of this study was to evaluate possible pathways through which social support may reduce cardiovascular disease risk. Baseline data from The National Heart, Lung, and Blood Institute Family Intervention Trial for Heart Health were included in this analysis (N = 501, mean age 48 +/- 13 years, 66% female, 36% nonwhite). Social support was calculated using the Enhancing Recovery in Coronary Heart Disease Patients Social Support Instrument, a validated tool measuring emotional and instrumental support. Emotional support is defined as the belief that one is cared about (high = score > or = 18); instrumental social support is defined as help with services (high = score > or = 3). Demographics, physical activity, and diet were assessed by standardized questionnaires, and cardiovascular disease risk factors were measured systematically. Linear regression models were adjusted for age, race/ethnicity, sex, marital status, and education. Higher instrumental social support was positively associated with minutes of physical activity per week (P = .007). Higher emotional social support was positively associated with number of days of physical activity per week (P = .023), number of servings of wine per week (P = .007), and increased high-density lipoprotein cholesterol (P = .014). A mediational analysis was performed, and the relationship between emotional social support and high-density lipoprotein cholesterol was significantly attenuated by physical activity days per week and number of servings of wine per week. No significant associations were found for other potential mediators tested including body mass index, waist circumference, and intake of omega-3 fatty acids, beer, or liquor. In this ethnically diverse population, emotional social support was linked to higher high-density lipoprotein cholesterol levels through increased physical activity and wine intake, suggesting possible mechanisms through which social support may reduce cardiovascular disease risk. PMID- 18382250 TI - American Heart Association: council on cardiovascular nursing department. PMID- 18382251 TI - Foreword: state of the science: physiological evaluation of cardiovascular status. PMID- 18382252 TI - Arterial pressure-based stroke volume and functional hemodynamic monitoring. AB - Arterial pressure-based methods are less-invasive methods used to measure stroke volume and to predict fluid responsiveness. An understanding of the assumptions of the measurements and clinical factors that affect their accuracy and ability to predict fluid responsiveness is imperative when deciding when and how to use these new technologies. Frequently asked questions about these technologies and the data provided are addressed. PMID- 18382253 TI - Pulmonary artery catheters: state of the controversy. AB - Since 1970, pulmonary artery catheters (PACs) have been used in clinical practice to monitor the hemodynamic status of critically ill and injured patients. This technology was introduced and commercialized without considerable testing to determine safety and efficacy. After years of common clinical use, investigators identified potential increases in mortality associated with PAC use. For the past decade, investigators have studied various patient populations to elucidate the safety and efficacy of the PAC. This article reviews the historical context of PAC use, findings from recent clinical trials intended to determine safety and efficacy, issues with reliability and validity of PAC use, and complications associated with PAC use. Data from recent clinical trials do not support routine use of PACs, and the authors suggest that PAC-guided therapy should be the focus of study in future trials. PMID- 18382255 TI - Biomarkers in acute cardiovascular disease. AB - Cardiovascular disease today remains a formidable foe affecting 1 in 3 Americans. The emergence of cardiac biochemical markers has provided clinicians unique insight into the state of the myocardium. In fact, cardiac biomarkers now represent an essential criterion in the definition of acute myocardial infarction. There has been impressive development of efficient and reliable assays to detect biomarkers in the serum. Together with patient history and electrocardiographic analysis, the invaluable information gained from serum cardiac biomarkers supports diagnosis, therapy selection, and determination of prognosis. Biomarkers such as troponin and creatine kinase MB have received well deserved attention for their ability to detect myocardial ischemia. Clinicians today use cardiac markers to identify ischemia as well as alternate clinical states. B-type natriuretic peptide, for instance, reflects myocardial stretch as seen in heart failure exacerbations and may well have promising prognostic significance. The purpose of this review is to discuss current and emerging cardiac biomarkers in acutely ill patients. The advantages and disadvantages of biomarkers will also be presented in the context of their clinical uses. Present markers are highly sensitive and specific to myocardial injury; however they do not specifically identify the method of injury. An exciting potential exists for future biomarkers to demonstrate enhanced specificity and earlier detection of compromised myocardium. PMID- 18382256 TI - Lower extremity venous disorders: implications for nursing practice. AB - Lower extremity chronic venous disorders are significant and common causes of vascular morbidity and mortality worldwide. Venous ulcer relapse rates are as high as 72% and account for approximately 80% to 90% of all lower extremity ulcers. Venous disorders, much like arterial disease, are often progressive and chronic in nature, resulting in enormous economic and human costs in terms of treatment, pain, and suffering. Individuals with skin changes are at high risk of recurrent leg ulcers, which negatively affect the quality of life for both the individual and the family. Prevention efforts should target anatomical vein dysfunction, edema, and skin changes to prevent leg ulcers. This article presents an overview of both clinical and laboratory assessments of lower extremity chronic venous disorders, including descriptions of the risks and pathogenesis for differential diagnosis and implications for nursing. PMID- 18382257 TI - Evaluation of lower extremity arterial circulation and implications for nursing practice. AB - Lower extremity arterial disease (LEAD) affects millions of Americans, causing impaired function and a high risk of nonhealing wounds, infection, and limb loss. This chronic, progressive condition is often silent until a life- or limb threatening complication occurs and impacts the quality of life of individuals and their families and results in high healthcare costs. Assessment forms the cornerstone of effective nursing care, yet despite several national guidelines, LEAD remains unrecognized in half of persons with the condition. This article presents an overview of assessment of LEAD, including descriptions of the risks, pathogenesis, and common characteristics of arterial disease. PMID- 18382258 TI - Biobanks and biomarker research in cardiovascular disease. AB - PURPOSE: The purpose of this article is to describe biobank processes and the sociocultural and nursing challenges of these processes. The article will present ways that thinking genetically can challenge cardiovascular nurses and help in the development of biobanks for cardiovascular research. It will emphasize the importance of the contributions of nursing to the development of biobanks and biobanking research. CONCLUSIONS: The influence of cardiovascular nurses on the development of biobanks for research in cardiovascular disease will result in accelerated discoveries that will lead to innovative, safe, effective therapeutics (translational research and personalized healthcare). The challenge is to educate and encourage clinicians to think genetically and use biobanks for research. PMID- 18382259 TI - Using devices for physiologic monitoring in heart failure. AB - Heart failure (HF) is a complex and costly disease process associated with high morbidity and mortality. Implanted cardiac rhythm management devices are increasingly used in the HF population to provide therapies such as protection from sudden death and cardiac resynchronization therapy. Device-based diagnostic monitoring provides clinicians with information that can assist in identifying patients at risk for HF decompensation and subsequent hospitalization. This article will review the evidence for using diagnostic information from cardiac rhythm management devices in the management of HF patients. Future advanced monitoring devices will also be discussed. PMID- 18382260 TI - Electrocardiographic evaluation of cardiovascular status. AB - The electrocardiogram (ECG) is indispensable for the diagnosis and management of patients with a wide variety of cardiac and noncardiac diseases. The purpose of this paper is focused on recent research that used ECG, specifically the long-QT interval and microvolt T wave alternans, for the evaluation of life-threatening ventricular arrhythmias. Although remaining to be validated, QT prolongation along with other emerging electrocardiographic indices such as T wave morphology, T peak-to-T end time, or beat-to-beat QT variability may be sensitive indicators of malignant polymorphic ventricular tachyarrhythmia, torsade de pointes. Microvolt T wave alternans may provide important information in identifying a low risk group with left ventricular dysfunction who is unlikely to benefit from unnecessary prophylactic implantable cardioverter defibrillator therapy. These ECG markers have the potential to aid in the safe administration of individualized medications, avoidance of sudden cardiac death, and provision of a noninvasive strategy to identify patients who are most and least likely to benefit from expensive prophylactic implantable cardioverter defibrillator placement. PMID- 18382264 TI - JIM interview. PMID- 18382261 TI - Electrocardiographic predictors of sudden cardiac death. AB - Sudden cardiac death (SCD) is widespread and the most serious of the cardiac diseases, accounting for over half of cardiovascular mortality in adults in the United States, and nearly 1 in 3 of these patients does not report symptoms of cardiac disease before the sudden death. Quantifying the left ventricular ejection fraction is currently the best way to risk-stratify patients for SCD and identify those who are most likely to benefit from the insertion of an implantable cardiac defibrillator (ICD). The strategy of systemically placing ICDs in patients at risk of SCD is expensive and leads to substantial psychological hardship. However, noninvasive electrocardiographic indices of depolarization and repolarization may better identify patients who are at an increased risk of SCD. Therefore, developing an approach to identify electrocardiographic changes associated with the highest risk of arrhythmic death could markedly improve patient selection for ICD therapy. This report describes electrocardiographic parameters that may be useful in identifying patients at risk of SCD. The state of the science currently suggests that it is unlikely that a single electrocardiographic parameter will predict SCD, but rather a risk stratification algorithm based on a combination of electrocardiographic parameters may yield the best result. PMID- 18382265 TI - Clopidogrel use in coronary heart disease and percutaneous coronary intervention. AB - Antiplatelet therapy has proven efficacy in the management of atherothrombosis. Clopidogrel, a thienopyridine, is a potent antiplatelet agent that achieves its antiplatelet effects by inhibiting the binding of adenosine 5' diphosphate to its platelet receptor. Large clinical trials have demonstrated a role for clopidogrel in the management of symptomatic atherosclerosis, acute coronary syndromes, and patients undergoing percutaneous coronary intervention. In this review, we discuss the pharmacology of clopidogrel including the mechanism of action, review the major clinical trials that have defined the current role of clopidogrel in coronary heart disease and percutaneous coronary intervention, and, finally, examine the concept of clopidogrel resistance and its potential clinical implications. PMID- 18382267 TI - Effects of additive therapy with spironolactone on proteinuria in diabetic patients already on ACE inhibitor or ARB therapy: results of a randomized, placebo-controlled, double-blind, crossover trial. AB - INTRODUCTION: Aldosterone seems to have deleterious effects on the kidneys. Many animal studies and a few clinical trials have shown that suppression of aldosteroneby aldosterone receptor blockers ameliorates these effects. METHOD: In a double-blind crossover study, patients with diabetic nephropathy who were already receiving either angiotensin converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB) were given spironolactone or matching placebo with 1 month of washout in between. Blood pressure (BP), serum creatinine, serum potassium, and spot urine protein/creatinine were measured at the beginning and end of each study period. RESULTS: Mean systolic BP on spironolactone went down from 153.64 (+/-25.95) at the beginning to 141.60 (+/-16.54) at the end of study (P = 0.01). Diastolic BP during spironolactone therapy did not change significantly. The urine protein/creatinine increased from 1.24 (+/-1.13) to 1.57 (+/-2.13) on placebo (P = 0.35) and decreased from 1.80 (+/-1.78) to 0.79 (+/ 0.99) during spironolactone therapy (P = 0.004). In other words proteinuria increased by 24% during the placebo treatment period but decreased by half (57%) during the active treatment. Serum potassium increased from 4.29 (+/-0.47) to 4.64 (+/-0.55) during spironolactone therapy (P = 0.002), no significant change with placebo. Whereas serum creatinine did not change on placebo, it increased from 1.35 (+/-0.54) to 1.56 (+/-0.62) on spironolactone (P = 0.006). Glomerular filtration rate decreased from 61.91 (+/-23.4) to 53.94 (+/-23.58) on spironolactone (P = 0.0001) but not on placebo. CONCLUSIONS: Addition of a modest dose of spironolactone to a regimen of ACEI or ARB in patients with diabetic proteinuria causes further reduction in proteinuria and also lowers the systolic BP. As with ACEI or ARB, spironolactone modestly reduces the glomerular filtration rate and raises serum potassium. PMID- 18382266 TI - Altered kinetics of interleukin-6 and other inflammatory mediators during exercise in children with type 1 diabetes. AB - BACKGROUND: Leukocyte mobilization and secretions of cytokines, chemokines, and growth factors in children during exercise are necessary biochemical signals for physiological growth and long-term cardiovascular protection. Because of glycemic instability, altered exercise responses, particularly the proinflammatory cytokine interleukin (IL)-6, may occur in type 1 diabetes mellitus (T1DM) that could influence the onset/progression of diabetic vascular complications. Relatively little is known, however, on most molecular aspects of immunomodulatory adaptation to exercise in diabetic children. METHODS: We therefore studied 21 children (age, 13.4 +/- 0.3 years; 13 boys/8 girls) with T1DM and 21 age-matched healthy controls during 30 minutes of intense and intermittent cycling exercise. Euglycemia was maintained during and for greater than 90 minutes before exercise; blood samples for IL-6 and other cytokines/chemokines were drawn before, during (every 6 minutes), and after (every 15 minutes) exercise. RESULTS: In T1DM, exercise-induced IL-6 peak occurred earlier and with greater magnitude than that in controls; an exploratory analysis of additional inflammatory mediators displayed a similarly accelerated/exaggerated pattern in T1DM, including the kinetic profiles of tumor necrosis factor alpha, IL-4, IL-12p70, IL-17, granulocyte-monocyte colony stimulating factor, monocyte chemoattractant protein-1, macrophage inflammatory protein-1alpha, and eotaxin (interferon-inducible protein-10 was the only measured variable essentially indistinguishable between groups). CONCLUSION: Therefore, during intense and intermittent exercise, significant alterations in the immunologic pattern of inflammatory regulation occurred in children with T1DM as compared with healthy controls. Our findings underscore how the understanding of all the underlying molecular mechanisms is a necessary prerequisite for achieving effective use of exercise and the full manifestation of its health benefits, particularly in understudied populations such as children with T1DM who are at increased risk for cardiovascular complications. PMID- 18382268 TI - Central role of rho kinase in lipopolysaccharide-induced platelet capture on venous endothelium. AB - BACKGROUND: Platelet-endothelial cell interactions play a key role in hemostasis and pathological coagulation and are dependent on different surface molecules that are expressed upon activation (eg, mediated by lipopolysaccharide [LPS]). Recently, we have shown that Rho kinase plays a central role in LPS-mediated leukocyte-endothelial cell interactions. We investigated the role of Rho-kinase in inflammatory interactions between platelets and the endothelium in femoral veins in vivo. METHODS: Mice were injected intravenously with LPS (0.5 mg/kg)/D galactosamine (900 mg/kg), and Rho kinase was blocked with fasudil 15 minutes before LPS application. Four hours after LPS administration, intravital fluorescence microscopy of the femoral vein was performed, and primary and secondary platelet-endothelial cell interactions were visualized after in vivo platelet staining with rhodamine 6G. RESULTS: Intravital microscopy showed a significant increase in platelet tethering, rolling, and firm adhesion as well as platelet secondary capture in LPS-treated mice. Rho-kinase inhibition by fasudil significantly reduced platelet tethering, rolling, and firm adhesion. Interestingly, functional blockade of Rho kinase was also able to diminish secondary platelet capture by 79%. CONCLUSIONS: From our data, we conclude that Rho-kinase signaling plays a central role in the regulation of LPS-induced platelet-endothelial cell interactions in large veins in vivo. Thus, Rho-kinase inhibition might be useful in prevention or treatment of pathological inflammation and endotoxin-mediated hypercoagulation. PMID- 18382269 TI - Retraction. PMID- 18382272 TI - Thalamus surface shape deformity in obsessive-compulsive disorder and schizophrenia. AB - The authors performed a three-dimensional shape deformation analysis to clarify the various patterns of specific thalamic nuclei abnormality using three age matched and sex-matched groups of 22 patients with obsessive-compulsive disorder (OCD), 22 patients with schizophrenia and 22 control participants. Compared with the healthy volunteers, the anterior, lateral outward surface deformities of the thalamus were significant in OCD patients, whereas the posterior, medial outward deformities of the thalamus were prominent in schizophrenia patients. In terms of thalamic asymmetry, both OCD and schizophrenia patients exhibited the loss of a leftward pattern of asymmetry on the posterior, medial surface of the thalamus. Different patterns of shape abnormality of specific thalamic nuclei may be related to the different phenomenology of OCD and schizophrenia. PMID- 18382271 TI - Multivariate analyses suggest genetic impacts on neurocircuitry in schizophrenia. AB - We investigated the relationship of functional neurocircuitries and dopamine receptor D1 (DRD1) polymorphisms in schizophrenics during a working memory task. Participants performed the Serial Item Recognition Paradigm memory task during functional magnetic resonance imaging acquisition. We performed a data-driven multivariate analysis (partial least squares) to characterize brain network (covariance) patterns. Genetic testing identified two main genotypes. Accuracy did not differ between the groups. Covariance patterns of different areas (including the dorsolateral prefrontal cortex and the inferior parietal lobule) were inversely related between the two genotypes. Two groups of schizophrenic patients with similar symptomatology and performance on a working memory task, but with distinct dopamine receptor genotypes, may use distinct neural systems to retrieve information. PMID- 18382273 TI - Amyloid-beta E22Delta variant induces synaptic alteration in mouse hippocampal slices. AB - We recently identified a novel amyloid precursor protein mutation (E693Delta) in familial Alzheimer's-type dementia. This mutation produces amyloid-beta (Abeta) variant lacking glutamate-22 (E22Delta), which showed enhanced oligomerization but no fibrillization. Here, we examined in-vitro toxicity of Abeta E22Delta peptide. Wild-type Abeta1-42 showed a dose-dependent (1 nM to 1 microM) cytotoxicity to cultured neuronal cells in the 3-(4,5-dimethylthiazol-2yl)-2,5 diphenyl tetrazolium bromide assay, whereas Abeta1-42 E22Delta was toxic only weakly at 1 microM. In mouse hippocampal slices, however, Abeta1-42 E22Delta caused a dose-dependent (0.1-10 microM) decrease of synaptophysin, whereas wild type Abeta1-42 was trophic at 0.1-1 microM and toxic at 10 microM. These results suggest that extracellular Abeta E22Delta causes more potent synaptic alteration, but lower neurodegeneration, than wild-type Abeta probably because of its unique aggregation property. PMID- 18382274 TI - Neuron-specific and inducible recombination by Cre recombinase in the mouse. AB - To investigate the neuronal function of genes in vivo, a neuron-specific and inducible gene targeting system is desirable. In this study, we generated a knockin mouse line that expresses a fusion protein consisting of the Cre recombinase and the progesterone receptor (CrePR) in neurons. The neuron-specific expression of CrePR was attained by inserting CrePR gene into the tau locus, because tau is expressed strongly in neurons but scarcely in glias and other tissues. By crossing this knockin mouse line (tau(CrePR)) with ROSA26 lacZ reporter mouse line (R26R), we observed that the antiprogesterone RU486 could induce recombinase activity of the CrePR specifically in neurons. Thus, tau (CrePR) knockin line is a useful tool for studying neuronal gene functions. PMID- 18382275 TI - Cortical activation during executed, imagined, and observed foot movements. AB - Evidence suggests that executed, imagined, and observed movements share neural substrates, however, brain activation during the performance of these three tasks has not yet been examined during lower extremity movements. Functional MRI was performed in 10 healthy right-footed participants during imagined, executed, and observed right ankle movements. Task compliance was high, confirmed via behavioral assessment and electromyographic measurements. Each task was also associated with its own profile of regional activation, however, overall, regional activation showed substantial overlap across the three lower extremity motor tasks. The findings suggest the utility of continued efforts to develop motor imagery and observation programs for improving lower extremity function in a range of clinical settings. PMID- 18382276 TI - Correlations in spontaneous activity and gray matter density between left and right sensoritmotor areas of pianists. AB - Resting-state functional MRI and structural MRI were used to study correlations of spontaneous activity and gray matter density between the left and right primary sensorimotor areas in pianists and nonmusicians. Functional MRI analysis showed significant correlation of spontaneous activity between the left and right primary sensorimotor area in both groups; however, there was no between-group difference. Structural MRI analysis showed significant correlation in gray matter density between the left and right sensorimotor areas in nonmusicians (r=0.65, P=0.001), but not in pianists (r=0.07, P=0.78), with a significant between-group difference (P=0.035). The lack of correlation of gray matter density between the left and right sensorimotor areas might be the basis of bimanual coordination of the pianists. PMID- 18382277 TI - Hypoxia-induced hypomyelination in the developing brain is mammalian target of rapamycin-4E-binding protein-1 signaling dependent. AB - We hypothesized that changes in the expression levels of genes in the mammalian target of rapamycin are involved in the hypoxia-induced growth retardation in the brain including hypomyelination. Microarray and proteomic studies showed a 2.3 fold increase in the expression levels of eukaryotic translation initiation factor 4E-binding protein-1 and a 3-fold decrease in the levels of FK506-binding protein-1 in a neonatal model of hypoxia, indicating a signal transduction impairment through mammalian target of rapamycin (mTOR). Analysis of hypoxic brain showed a marked decrease in the phosphorylation levels of 4E-binding protein-1, suggesting a reduction of mTOR activity. These data suggest that suppression of mTOR may be the mechanism underlying hypoxia-induced hypomyelination observed in the developing brain. PMID- 18382278 TI - Magnetoencephalographic study of the components of event-related fields in object recognition. AB - Neuromagnetic fields were recorded from normal participants to study neural activation while they performed a delayed matching-to-sample task of visual stimuli. Each participant was asked to compare sequentially presented three dimensional object images under same view and different view conditions. Minimum current estimates were calculated to characterize cortical activities for the object recognition. Activation was observed in the bilateral parieto-occipital regions at approximately 180 ms. Activation within the right parieto-occipital region was significantly enhanced under the different view conditions. This result indicates that the activity at the right parieto-occipital region is related to viewpoint and may play an important role in three-dimensional object recognition. PMID- 18382279 TI - Neuroprotection against ischemic brain injury by knockdown of hematopietic progenitor kinase 1 expression. AB - In this study, we investigated the role of hematopietic progenitor kinase 1 (HPK1) in delayed neuronal damage after cerebral ischemia and the possible regulatory mechanisms of this event. Our data show that tyrosine phosphorylation of HPK1 was significantly increased at 6 h of ischemic-reperfusion compared with sham control. The increase in p-HPK1, p-MLK3, p-MKK7, and p-JNK3 was attenuated by HPK1 antisense oligodeoxynucleotides intra-cerebroventricular infusion, but not MS-ODNs or vehicle. Intracerebroventricular infusion of antisense oligodeoxynucleotides also increased the number of surviving pyramidal neurons, whereas MS-ODNs or vehicle (TE) groups had no effects. These results indicate that knockdown of HPK1 expression provides neuroprotection through downregulation activation of the MLK3-MKK7-JNK3 pathway following cerebral ischemia in the rat hippocampus CA1 subfield. PMID- 18382280 TI - Neuroanatomic correlates of autism and stereotypy in children with Down syndrome. AB - We conducted semiautomated, atlas-based analyses of regional brain volume changes on MRIs of children and adolescents with Down syndrome (DS) (N=15), DS with comorbid autism spectrum disorder (ASD) (N=15), and age-matched or sex-matched typically developing controls (N=22). Selective volumetric changes were correlated with neurobehavioral measures to determine their functional significance. DS involved selective reduction of frontal and parietal gray matter volumes, beyond the global microencephaly typically observed in this condition. DS with comorbid ASD involved relative hyperplasia of white matter in the cerebellum and brainstem compared with DS only. Cerebellar white matter volumes were positively correlated with severity of stereotypies, a distinctive feature of ASD in DS. PMID- 18382281 TI - The novel cyclooxygenase-2 inhibitor GW637185X protects against l-methyl-4-phenyl 1,2,3,6-tetrahydropyridine toxicity. AB - The possible neuroprotective role of a novel and highly selective cyclooxygenase 2 inhibitor GW637185X was studied in a model of acute 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-induced injury of nigrostriatal dopaminergic (DA) neurons in the mouse. Stereological and microdensitometrical analysis of nigral tyrosine hydroxylase-immunoreactive cell bodies and striatal tyrosine hydroxylase immunoreactive terminals, respectively, showed that GW637185X exerted a full protection against MPTP-induced degeneration of the nigro-striatal pathway. In contrast to earlier studies, these findings demonstrate that acute inhibition of cyclooxygenase-2 can result in a full neuroprotective effect not only on nigral DA cell bodies, but also on striatal DA terminals in the mouse MPTP model. PMID- 18382282 TI - Deep brain stimulation promotes excitation and inhibition in subthalamic nucleus in Parkinson's disease. AB - Subthalamic nucleus single-unit recordings were undertaken before and during pedunculopontine [corrected] nucleus-stimulation at clinically relevant frequency (25 Hz) in six patients with Parkinson's disease. Pedunculopontine [corrected] nucleus stimulation changed the firing activity of almost every subthalamic nucleus cell (44/48) by decreasing the ongoing discharge in bursting subthalamic nucleus neurons (-62.1%) and exciting irregular (+63.2%) and tonic/regular discharges (+20.1%). These conflicting data challenge the definition of pedunculopontine [corrected] nucleus as a new target area for Parkinson's disease. If the modulation of subthalamic nucleus bursting units may corroborate the pedunculopontine [corrected] nucleus therapeutic role, the simultaneous excitatory influence during nonbursty patterns might interfere with a favorable outcome on motor signs. As a result, the implantation of pedunculopontine [corrected] nucleus alone may be hazardous while the association of subthalamic nucleus plus pedunculopontine [corrected] nucleus seems to be reasonable. PMID- 18382283 TI - Inactivation of the lateral septum blocks fox odor-induced fear behavior. AB - A number of lesion studies have shown that the lateral septum plays an important role in the modulation of innate fear. Furthermore, an increased c-fos expression in the lateral septum was demonstrated after exposure to natural predator odors and 2,3,5,-trimethyl-3-thiazoline (TMT), a component of fox odor. This study investigates, on a behavioral level, whether the lateral septum plays a role in TMT-induced fear. Temporary inactivation of the lateral septum by local muscimol injections clearly blocked TMT-induced fear behavior but had no effect on behavior in a controlled condition. This indicates that the lateral septum is important for the processing of TMT-induced fear and suggests that the lateral septum is also involved in fear behavior induced by natural predator odors. PMID- 18382284 TI - Different responses to same stimuli. AB - Identical comparison stimuli were induced to be recognized as schematic faces or the figures with three English letters 'O' (EngO) by verbal bias and two different contextual priming conditions. Our findings confirmed that there might be a coarse visual categorization around 40-100 ms at mid-parietal site. Larger P1 responses to schematic faces compared with EngO definitely revealed that P1 at occipito-temporal sites should be thought to represent an earlier face-specific processing stage, as early as around 100-120 ms after stimulus onset rather than simply reflect the low-level visual difference. A significant difference of N170/vertex positive potential between the identical comparison stimuli reinforced the evidence that they were early markers of face processing and corroborated the unequivocal influence of contextual effect. PMID- 18382285 TI - Intracellular calcium concentration changes initiated by N-methyl-D-aspartic acid receptors in retinal horizontal cells. AB - Intracellular calcium concentration changes initiated by N-methyl-D-aspartic acid receptors were studied in carp retinal horizontal cells. Fura-2 fluorescent calcium imaging showed that H1 subtype horizontal cells responded to exogenously applied N-methyl-D-aspartic acid with a transient intracellular free Ca(2+) ([Ca2+]i) increase that decayed to a sustained, but elevated level of [Ca(2+)]i. Contributions of different Ca(2+) flux pathways underlying the time course of this increase in [Ca(2+)]i were explored via experiment as well as via a computational model based on the biophysical properties of H1 cells. Intracellular calcium stores were suggested to play crucial role in the initial transient increase of [Ca(2+)]i. PMID- 18382286 TI - Functional evidence for internal feedback in the songbird brain nucleus HVC. AB - The song control system of songbirds consists mainly of the 'motor pathway' and 'anterior forebrain pathway'. The medial magnocellular nucleus of anterior nidopallium (mMAN) projects to the song control nucleus HVC, which is the point of divergence of the two pathways. We made simultaneous multiunit electrophysiological recordings from the mMAN and HVC in anesthetized Bengalese finches. We confirmed that the mMAN neurons showed song-selective auditory responses, and found temporal correlations between song-related activities of the mMAN and HVC neurons. The temporal relationship between the neural activation of the HVC and mMAN suggests that these nuclei are parts of a closed loop, which could provide internal feedback to the HVC for sequential syllable control. PMID- 18382287 TI - Recovery of nerve injury-induced alexia for Braille using forearm anaesthesia. AB - Nerve injuries in the upper extremity may severely affect hand function. Cutaneous forearm anaesthesia has been shown to improve hand sensation in nerve injured patients. A blind man who lost his Braille reading capability after an axillary plexus injury was treated with temporary cutaneous forearm anaesthesia. After treatment sensory functions of the hand improved and the patient regained his Braille reading capability. The mechanism behind the improvement is likely unmasking of inhibited or silent neurons, but after repeated treatment sessions at increasing intervals the improvement has remained at 1-year follow-up, implying a structural change in the somatosensory cortex. PMID- 18382288 TI - Transcription factor assembly on the nicotinic receptor beta4 subunit gene promoter. AB - Nicotinic acetylcholine receptors are involved in a plethora of fundamental biological processes ranging from muscle contraction to formation of memories. The receptors are pentameric proteins whose subunits are encoded by distinct genes. Subunit composition of a mature nicotinic receptor is governed in part by the transcriptional regulation of each subunit gene. Here, using chromatin immunoprecipitation assays, we report the interaction of the transcription factors Sp1, Sp3, c-Jun and Sox10 with the beta4 subunit gene promoter in neuronal-like cell lines and rodent brain tissue. Our results corroborate previous in-vitro data demonstrating that these transcription factors interact with the beta4 promoter. Taken together, these data suggest that Sp1, Sp3, c-Jun and Sox10 regulate expression of the beta4 subunit gene in the mammalian brain. PMID- 18382289 TI - Reduced self-motion perception in patients with midline cerebellar lesions. AB - Vestibular input to the cerebellum mediates balance and eye movement control. Recent functional MRI studies, however, show midline cerebellar activation during visually induced illusions of self-rotation, thus suggesting that the cerebellum may also contribute to self-motion perception. Here, we investigate self-motion perception directly in patients with vermal (or midline) cerebellar ataxia. Participants were rotated in the dark (90 degrees /s velocity steps) and the time constant of decay of the postrotational angular velocity sensation was measured. The perceptual vestibular time constant in patients was considerably reduced (7.8 s) with respect to control values in this (25.6 s) and several previous studies. In addition to the processing of vestibular signals for motor control, the cerebellar vermis is involved in vestibular processing of self-motion perception. PMID- 18382290 TI - Semantic composition engenders an N400: evidence from Chinese compounds. AB - This study provides evidence for the role of semantic composition in compound word processing. We examined the online processing of isolated two meaning unit compounds in Chinese, a language that uses compounding to 'disambiguate' meaning. Using auditory presentation, we manipulated the semantic meaning and syntactic category of the two meaning units forming a compound. Event-related brain potential-recordings revealed a significant influence of semantic information, which was reflected in an N400 signature for compounds whose meaning differed from the constituent meanings. This finding suggests that the combination of distinct constituent meanings to form an overall compound meaning consumes processing resources. By contrast, no comparable difference was observed based on syntactic category information. Our findings indicate that combinatory semantic processing at the word level correlates with N400 effects. PMID- 18382292 TI - Juvenile intradural chordoma: case report. AB - OBJECTIVE: We report the youngest known case of a prepontine intradural chordoma. These tumors are exceedingly rare. Unlike their more common extradural counterparts, no recurrence of an intradural chordoma has been reported. CLINICAL PRESENTATION: A 9-year-old boy underwent diagnostic imaging for evaluation of headaches. Although neurologically intact, a magnetic resonance imaging scan revealed a large prepontine mass with focal enhancement. INTERVENTION: Endoscopic assisted gross total resection was attained with staged bilateral retrosigmoid approaches. There were no additional adjuvant therapies. At the time of the 1 year follow-up evaluation, the patient had no recurrence. CONCLUSION: By using an endoscopic-assisted procedure, we achieved complete resection of an intradural chordoma offering a potential for surgical cure. Resection is particularly advantageous because it spares the young child the need for radiation treatment. Close follow-up is warranted because we postulate that this tumor exists in a biological continuum between benign notochordal hamartomatous remnants and typical invasive chordomas. PMID- 18382293 TI - Bilateral osteomas arising from the internal auditory canal: case report. AB - OBJECTIVE: Osteomas arising from the internal auditory canal and developing in the cerebellopontine angle have rarely been reported. We present the first case of bilateral osteomas in this region and describe our management strategy. CLINICAL PRESENTATION: A 30-year-old woman presented with strong vertigo, tinnitus, and hypacusis on the left side. Brain magnetic resonance imaging and computed tomographic scans displayed bilateral cerebellopontine angle osteomas arising from the internal auditory canal. The larger tumor on the left side was found to be compressing the vestibulocochlear nerve. INTERVENTION: Surgery was performed on the symptomatic side via the retrosigmoid approach, and the tumor was removed completely. The vertigo resolved completely after surgery, and the patient had no further tinnitus attacks. An audiogram showed slightly improved hearing with a mean of 20 dB in the main speech area. CONCLUSION: Osteomas should be considered in patients with bilateral cerebellopontine angle tumors. Surgical removal might provide resolution of symptoms. PMID- 18382294 TI - Postmortem study of deep brain stimulation of the anterior thalamus: case report. AB - OBJECTIVE: As new clinical applications for deep brain stimulation (DBS) emerge and the number of patients with DBS systems continues to grow, lead technology will also advance. To direct improvement of these leads, improved understanding of the effects of the DBS electrodes and stimulation parameters on the surrounding brain parenchyma is necessary. We present a postmortem evaluation of a patient who had previously undergone bilateral DBS of the anterior thalamic nucleus. CLINICAL PRESENTATION: A 21-year-old, right-handed man with a 2-year history of epilepsy secondary to encephalitis underwent bilateral DBS of the anterior nucleus of the thalamus. He died 8 months after surgery, and his death was classified as a sudden, unexpected, unexplained death as a result of epilepsy. INTERVENTION: Microscopic examination and immunohistochemical analysis using glial fibrillary acidic protein, CD11b, CD45, and CD 68 were performed. The thalami of this patient were then compared with brain tissue obtained from a 45 year-old patient who died as a result of a myocardial infarction and had no history of neurological disease and no surgical intervention. There were no differences in the microscopic and histochemical evaluation of the thalami between patients, other than immediately around the electrode tract. Minimal tissue damage, mild astrocytosis, and mild inflammation surrounding the electrode termination site were observed. CONCLUSION: We report the first postmortem examination after bilateral DBS of the anterior nucleus of the thalamus for epilepsy. A comparison with control tissue showed no significant difference other than mild inflammation along the lead track. PMID- 18382295 TI - Diplopia after balloon compression of retrogasserian ganglion rootlets for trigeminal neuralgia: technical case report. AB - OBJECTIVE: Balloon compression of the rootlets behind the trigeminal ganglion for the treatment of trigeminal neuralgia has become an increasingly popular method among neurosurgeons. However, the method has recognized complications, including double vision. Although occurring infrequently, diplopia may cause the patient significant disability. To minimize the risk for this complication, we analyzed our patients with respect to the surgical technique. METHODS: We reviewed our joint consecutive series of 193 patients with trigeminal neuralgia treated with balloon compression. The medical records and the intraoperative x-ray images were analyzed. RESULTS: We identified six patients with double vision postoperatively. In analyzing these occurrences, we found that the balloon was inflated outside Meckel's cave in four patients, the balloon was initially inflated too deeply in one patient, and the anatomy of Meckel's cave was probably aberrant in one patient. In five of the six patients, the symptoms resolved within 5 months. CONCLUSION: By meticulous surgical technique with close attention to the anatomic position and the shape of the inflated balloon, most cases of postcompression diplopia should be avoided. PMID- 18382306 TI - Neurosurgery in the realm of 10(-9), Part 2: applications of nanotechnology to neurosurgery--present and future. AB - Neurosurgery in the future will witness an increasing influx of novel technologies, many of which will be based on developments in the emerging science of nanotechnology. Additionally, the continued trend in medicine toward minimally invasive diagnostic and surgical techniques will be aided by incorporation of applications of nanotechnology. Neurosurgeons of the future must facilitate the development of nanotechnology and nanomedicine in their clinical practice and research efforts to optimize patient benefit and facilitate scientific advancement. The fields of nanotechnology and nanomedicine remain in their infancy. Recently, however, the literature regarding nanoscience has rapidly expanded. This article is the second of two and provides a review of recent nanotechnology research relevant to clinical neurosurgery and neurology. The first article reviewed recent developments and issues in nanotechnology with a particular focus on applications to the neurosciences. This article also discusses current developments in nanotechnology and nanomedicine that may yield applications in neurosurgery in the future. Additional attention is given to other emerging technologies that are not truly nanotechnology, such as microelectromechanical systems, which will influence the future of medicine and neurosurgery. The goal is to provide the reader with a brief outline and description of some of the new developments in nanotechnology that may affect the clinical practice or operative experience of neurosurgeons. Continued innovation in nanotechnology presents novel opportunities for translation to the clinical arena. Neuroscience, neurology, and neurosurgery will be greatly affected by the influx of nanoscience and its applications. Through continued collaboration, physicians, scientists, and engineers will shape the futures of nanomedicine and nanoneurosurgery. PMID- 18382307 TI - An image-guided magnetic resonance-compatible surgical robot. AB - OBJECTIVE: The past decade has witnessed the increasing application of robotics in surgery, yet there is no existing system that combines stereotaxy and microsurgery in an imaging environment. To fulfill this niche, we have designed and manufactured an image-guided robotic system that is compatible with magnetic resonance imaging. METHODS: The system conveys the sight, touch, and sound of surgery to an operator seated at a remote workstation. Motion scaling, tremor filtering, and precision robotics allow surgeons to rapidly attain technical proficiency while working at a spatial resolution of 50 to 100 microm instead of a few millimeters. This system has the potential to shift surgery from the organ toward the cellular level. RESULTS: By integrating the robot with images obtained during the procedure, the effects of surgery on both the lesion and brain are immediately revealed. CONCLUSION: We are providing technology to advance and transform surgery with the potential to improve patient outcome. PMID- 18382308 TI - Comparative study of cranial topographic procedures: Broca's legacy toward practical brain surgery. AB - OBJECTIVE: In many locations, neurosurgeons still use stereotactic- or anatomic based craniocerebral topography systems to identify cortical landmarks. However, their predictive value for identifying two key landmarks--the central sulcus (CS) and lateral sulcus (LS)--has never been evaluated. We quantitatively compare leading craniocerebral topographic methods and review their historical significance for neurosurgery. METHODS: On 12 cadaveric head sides, the methods of Broca, Reid, Poirier, Taylor-Haughton, and Rhoton were used to predict positions of the CS and LS. After craniotomy, the actual CS, LS, and the superior and inferior Rolandic points were identified. Distances between predicted positions and actual structures were measured, and the systems were compared. RESULTS: The actual superior Rolandic point was 4.6 +/- 2.9 mm anterior to prediction by Broca's method; 4.3 +/- 2.13 mm anterior to the Poirier, Taylor Haughton, and Rhoton methods; and 3.26 +/- 3.17 mm anterior or posterior using Reid's method. The actual inferior Rolandic point was anteroinferior to all predictions: 5.87 +/- 3.1 mm by Rhoton, 6.97 +/- 3.55 mm by Broca, 7.64 +/- 2.54 mm by Poirier, and 7.61 +/- 3.85 mm by Reid and Taylor-Haughton. The actual LS was 2.33 mm away from the predicted point using Poirier's method, and 2.00 mm away from the predicted point using the Reid, Taylor-Haughton, and Rhoton methods. CONCLUSION: Predicting positions of the CS and LS to within a few millimeters, these landmark methods remain reliable for cerebral localization. Largely initiated by the work of Paul Broca, these systems lay at the development of a practical method of neurosurgery in the late 19th century. PMID- 18382309 TI - L-dopa-induced dyskinesia and stereotactic surgery for Parkinson's disease. AB - OBJECTIVE: To assess the impact of different surgical targets and techniques, such as ablation and deep brain stimulation, to treat patients with L-dopa induced dyskinesia (LID), a major therapeutic complication of Parkinson's disease. METHODS: This review analyzes the effects of early surgical procedures to treat hyperkinesia and the current methods and targets used to combat LID in Parkinson's disease, which are mainly thalamotomy, pallidotomy, and deep brain stimulation of the globus pallidus internus and the subthalamic nucleus. RESULTS: Available information indicates that surgery of the globus pallidus internus and thalamus (the pallidal receiving area) and of the subthalamic nucleus has a pronounced antidyskinetic effect. This effect is associated with a concomitant improvement in the parkinsonian ("off"-medication) state. Although it is more profound with pallidal and subthalamic surgery, such an effect can also be observed to some extent with thalamic surgery. The latter is attributable to the fact that surgery of the ventralis intermedius is primarily effective for treating tremor. An integral pallidothalamic pathway is needed for dyskinesia to be expressed. Thus, LID is less frequent after subthalamotomy or deep brain stimulation of the subthalamic nucleus through a functional effect mediated by the physiological normalization of the motor system and by an indirect effect associated with a reduction in the daily dose of L-dopa. CONCLUSION: Surgery is the only treatment available for Parkinson's disease that can predictably improve both the parkinsonian motor syndrome and LID. The exact mechanisms involved in these effects are not well understood. Pallidal and thalamic surgery affecting pallidal relays reduce LID frequency by disrupting the pallidothalamic circuit, probably eliminating the neuronal activity associated with dyskinesia. Alternatively, the antidyskinetic effect of subthalamic nucleus surgery may in part be attributable to a reduction in the L-dopa dose as well as to the stabilization of the basal ganglia circuits after the surgical procedure. PMID- 18382310 TI - Long-term seizures and quality of life after epilepsy surgery compared with matched controls. AB - OBJECTIVE: We compared long-term seizure outcome and health-related quality of life (HRQoL) of patients who underwent epilepsy surgery and matched medically treated nonsurgical controls with intractable epilepsy. METHODS: Medically treated controls were identified for patients operated on for epilepsy between January 1, 1949 and December 31, 1992. We used a matched cohort design, matching for age, sex, and seizure type. The analysis was based on 70 complete matching pairs. HRQoL was assessed with the Quality of Life in Epilepsy Inventory 89 questionnaire an average of 15 years after surgery. RESULTS: Among surgery patients, 48% were seizure-free during the previous year compared with 19% of the controls (P = 0.0004). Fewer surgery patients used antiepileptic drugs (70%) than controls (93%). The odds of being seizure-free were higher for surgery patients in total and in subgroups divided according to length of follow-up. The mean HRQoL for surgery patients was higher in five of the 17 Quality of Life in Epilepsy Inventory 89 dimensions and worse in none. Among patients with more than 7 years of follow-up, HRQoL was better in three dimensions and worse in none. Among patients with 7 years of follow-up or less, HRQoL was better in two dimensions and worse in the language dimension of the Quality of Life in Epilepsy Inventory 89. CONCLUSION: After an average of more than 15 years of follow-up, epilepsy surgery patients had fewer seizures, used less antiepileptic medication, and had better HRQoL in several dimensions of the Quality of Life in Epilepsy Inventory 89 instrument than matched medically treated controls with refractory epilepsy, although possibly at a slight disadvantage in the language dimension among those with 7 years of follow-up or less. PMID- 18382311 TI - The clinical course after stereotactic radiosurgical amygdalohippocampectomy with neuroradiological correlates. AB - OBJECTIVE: Radiosurgical ablation of the mesial temporal lobe structures can be used in the treatment of intractable temporal lobe epilepsy associated with mesial temporal sclerosis. In this study, we analyzed the magnetic resonance imaging (MRI) and spectroscopic changes that follow the treatment and report the clinical sequelae of the procedure. METHODS: Eight patients (five men and three women; age, 38 +/- 15 yr [mean +/- standard deviation]) with mesial temporal sclerosis were treated with radiosurgical amygdalohippocampectomy (25 Gy to the 50% isodose region with a mean target volume of 6.2 +/- 0.7 cm). MRI and magnetic resonance spectroscopy were performed sequentially during a 24-month period after treatment. RESULTS: Patients were followed up clinically for 24 to 53 months. MRI scans revealed changes of marked temporal lobe swelling, with often markedly elevated apparent diffusion coefficients in keeping with vasogenic edema that became apparent 6 to 12 months after stereotactic radiosurgery. Spectroscopy of the target area revealed a progressive loss of N-acetylaspartate (the late evolution of lactate) and a peak in the choline-to-creatine ratio that seemed to coincide with the peak of the vasogenic edema in the temporal lobe surrounding the target area. Clinically, all patients showed some reduction in seizure frequency, although in two patients, this reduction was modest. The MRI changes in those patients were also modest, and three patients ultimately became free of seizures. However, there was a latency of 18 to 24 months before improvements in seizure control occurred, and during this period, seizures worsened or changed in four patients. Two patients also developed symptoms of increased intracranial pressure with mild dysphasia, which responded to administration of corticosteroid medication. However, no long-term clinical verbal memory decline was identified in any patient. CONCLUSION: There are marked changes in MRI scans and magnetic resonance spectroscopic findings after patients undergo radiosurgery for temporal lobe epilepsy. Our initial findings suggest that some patients may have a period of distressing symptoms that accompany changes that are visualized on the MRI scans. PMID- 18382312 TI - Bilateral electrical stimulation of prelemniscal radiations in the treatment of advanced Parkinson's disease. AB - OBJECTIVE: Tremor and rigidity have been efficiently controlled by electrical stimulation of contralateral prelemniscal radiations (Raprl) in patients with unilateral Parkinson's disease. The present study determines the effect of bilateral Raprl electrical stimulation in a group of patients with severe bilateral tremor, rigidity, and bradykinesia. METHODS: Five patients with Parkinson's disease (Hoehn and Yahr scale, Stage V) underwent bilateral stereotactic electrode implantation. Postoperative magnetic resonance imaging studies confirmed their position. Bipolar chronic electrical stimulation was performed through contiguous contacts of each electrode, which were selected by means of a screening test that explored multiple combinations. Preoperative and 3 , 6-, 9-, and 12-month postoperative evaluations were performed using international rating scales. Postoperative evaluations were performed with 24 hours off medication-on stimulation. RESULTS: Contralateral tremor and rigidity were significantly decreased by simple insertion of electrodes in Raprl and returned hours to days later. Contacts for chronic stimulation were located in the area between the red and subthalamic nuclei, including Raprl, zona incerta, and substantia Q. Efficient stimulation had at least one contact in Raprl and in four cases, both contacts were only in Raprl. Stimulation parameters were 90 to 130 Hz, 90 to 330 mus, and 1.5 to 3.5 V. Unified Parkinson's Disease Rating Scale (motor, Part III) scores decreased 65% (P < 0.001), with improvements of 90% in tremor (P < 0.001), 94% in rigidity (P < 0.001), 75% in bradykinesia (P < 0.001), 40% in gait, and 35% in postural stability (P < 0.05) at 1 year. CONCLUSION: Raprl electrical stimulation is safe and efficient to treat patients with the Parkinson's disease symptomatic triad. By use of Raprl stereotactic coordinates, electrodes were placed behind the subthalamic nucleus. PMID- 18382313 TI - Deep brain stimulator hardware-related infections: incidence and management in a large series. AB - OBJECTIVE: Device-related infection is a common complication of deep brain stimulator (DBS) implantation. We reviewed the incidence and management of early hardware-related infections in a large series. METHODS: All patients undergoing DBS implantation surgery between 1998 and 2006 at a single institution were entered into a prospectively designed database. After database verification by cross-referencing manufacturer implantation records, a query was performed to include all new Medtronic (Minneapolis, MN) implantations performed with standard operating room technique. Hardware-related infections requiring further surgery were identified, and charts were reviewed to assess the success of lead-sparing partial hardware removal in this group. RESULTS: Four hundred twenty patients received 759 new DBS electrodes and 615 new internal pulse generators for the treatment of movement disorders or pain. Nineteen patients (4.5%) had an early (<6 mo) hardware-related infection requiring further surgery. There were no intracranial infections. Four patients presented with extensive cellulitis or wound dehiscence and were treated with total hardware removal. Fourteen patients presented with more localized infections and were treated by removal of the involved components only, followed by intravenously administered antibiotics. In nine of these patients, partial hardware removal successfully resolved the infection without requiring removal of the DBS electrodes. Wound washout alone was attempted in one patient and failed. CONCLUSION: In a large series of new DBS hardware implantations, the incidence of postoperative hardware-related infection requiring further surgery was 4.5%. When only one device component was involved, partial hardware removal was often successful. PMID- 18382314 TI - Neurovascular compression anatomy and pain outcome in patients with classic trigeminal neuralgia treated by radiosurgery. AB - OBJECTIVE: To study the influence of the anatomy of neurovascular compression (NVC) on pain outcome in patients with classic trigeminal neuralgia treated by radiosurgery. METHODS: Analysis of the anatomy of the trigeminal nerve, brainstem, and vessels was performed in 89 consecutive patients treated by Leksell gamma knife (Elekta Instruments, Stockholm, Sweden) for classic trigeminal neuralgia. One-millimeter axial magnetic resonance imaging slices (T1 weighted, T1-weighted enhanced, and T2-weighted selected partial inversion recovery) with coronal, sagittal, and three-dimensional reconstructions were viewed. The end point for outcome was total pain remission and no medication. The follow-up period ranged between 6 and 42 months. RESULTS: In 82 patients (92%), a vascular structure in contact with the nerve was observed. In four patients, the NVC was produced by a large vessel (basilar or vertebral artery) and in 78 by a smaller vessel. The superior cerebellar artery was the cause of the NVC in 64 patients (78%). The NVC was proximal (<3 mm to the brainstem) in 34 patients (41%) and distal in 43 patients (52%). Nerve dislocation was present in 29 patients (33%), and nerve atrophy was present in 21 (24%) patients. Visualization of NVC on magnetic resonance imaging scans was not associated with outcome. The two variables associated with poor outcome were a large vessel contacting the nerve with brainstem deformation and proximal NVC. Nerve atrophy and nerve dislocation were not associated with outcome. CONCLUSION: The visualization of NVC, nerve atrophy, and nerve dislocation on magnetic resonance imaging scans was not associated with pain outcome. A large vessel compressing the nerve and deforming the brainstem and proximal NVC were associated with a lesser pain control. PMID- 18382315 TI - Ruptured intracranial dermoid cysts: clinical, radiographic, and surgical features. AB - OBJECTIVE: Intracranial dermoid cysts are pathologically characterized by a thick, stratified squamous epithelium cyst wall containing dermal elements. Rupture into the subarachnoid spaces and ventricles is extremely rare. We review the clinical, radiographic, and surgical features of eight ruptured dermoid cysts. METHODS: We retrospectively evaluated five surgically treated patients with pathologically proven ruptured dermoid cysts. Clinic notes, operative reports, and neuroimaging, including initial computed tomographic and magnetic resonance imaging scans, were reviewed. Imaging was also available on three outside patients reviewed by members of our radiology department. RESULTS: The most common presentations were headaches (57%) and seizures (42%), followed by hydrocephalus (29%) from intraventricular rupture. These lesions were consistently hypodense on computed tomographic scans and hyperintense on T1 weighted images with minimal to no enhancement after gadolinium administration. Disseminated fat droplets were present in the subarachnoid space in both cerebral hemispheres in all patients, and five patients had intraventricular rupture with fat-fluid levels in the ventricles. Gross to near-total resection of the primary lesion was achieved in all five surgically treated patients treated at our institution. Four patients had remnant tumor capsules adherent to neurovascular structures that were unresectable. Repeat resection was performed for one recurrence; there were no further recurrences during a follow-up period of 2 to 134 months (mean, 65.6 mo). Two patients with preoperative hydrocephalus eventually required ventriculoperitoneal shunting. CONCLUSION: Ruptured intracranial dermoid cysts represent 0.18% of all central nervous system tumors surgically treated in our institution during a 12-year period. The presence of disseminated fat droplets in the subarachnoid space or ventricles on neuroimaging is diagnostic for a ruptured dermoid cyst. Gross total removal is achievable; however, residual tumor capsules adherent to neurovascular structures should be left behind to minimize complications. PMID- 18382316 TI - Cognitive outcomes after carotid revascularization: the role of cerebral emboli and hypoperfusion. AB - OBJECTIVE: Carotid artery stenting (CAS) and carotid endarterectomy (CEA) are currently being compared in ongoing randomized, controlled trials using postprocedural 30-day stroke rate, myocardial infarction, and mortality as primary endpoints. Recent data suggest that cognitive function may decline after CEA. Understanding the mechanisms that affect cognitive outcomes after carotid revascularization will be important in the design of future comparative studies of CAS and CEA incorporating cognitive outcome as an endpoint. SUMMARY OF REVIEW: The effects of carotid revascularization procedures on cognitive outcome are unclear. Several factors contribute to the difficulty in interpreting cognitive data, including patient heterogeneity, variability of surgical techniques, and the differences in neuropsychological testing methodology. Mechanisms underlying cognitive effects during CEA have emerged, including the potential detrimental effect of procedural emboli and the beneficial effect of improved cerebral hemodynamics. The emergence of CAS as an alternative to CEA for treating carotid stenosis again raises questions about cognitive outcomes. Despite the use of distal protection devices, CAS is associated with a higher burden of microemboli. CAS does not, however, require the extent of temporary vessel occlusion associated with CEA. Quantifying microemboli and changes in cerebral hemodynamics along with standardization of neuropsychological testing may lead to meaningful comparisons of cognitive data for patients undergoing carotid revascularization procedures. CONCLUSION: As use of CAS increases, it is important for randomized, controlled trials comparing CAS with CEA to include cognitive outcomes assessments. Furthermore, understanding the key mechanisms resulting in cognitive impairment during carotid revascularization procedures might limit injury. PMID- 18382317 TI - Controversies in the surgical treatment of ruptured intracranial aneurysms: the First Annual J. Lawrence Pool Memorial Research Symposium--controversies in the management of cerebral aneurysms. AB - The management of aneurysmal subarachnoid hemorrhage has evolved over time, including the use of the microscope for aneurysm clip application, improved imaging modalities, endovascular methods for aneurysm treatment, dedicated neurointensive care units, and more aggressive therapy for cerebral vasospasm. Although these advancements have reduced the morbidity and mortality associated with aneurysmal subarachnoid hemorrhage, outcomes for this patient population continue to leave much room for improvement. This work highlights controversial adjuvant techniques, maneuvers, and therapies surrounding the surgical treatment of ruptured cerebral aneurysms that currently lack a consensus opinion. These treatments include centralized care in high-volume centers, as well as the use of antifibrinolytic therapy, routine cerebrospinal fluid diversion, intraoperative hypothermia, temporary clip application, neuroprotective drugs, intraoperative angiography, and decompressive hemicraniectomy. Although definitive answers will only be possible through future multicenter collaboration, we review the controversy surrounding these adjuncts and report the consensus opinion from a highly experienced audience. PMID- 18382318 TI - Transarterial embolization of clival dural arteriovenous fistulae using liquid embolic agents. AB - OBJECTIVE: Dural arteriovenous fistulae (DAVFs) rarely involve the clivus. This report examines the clinical presentation, angiographic findings, endovascular management, and outcome of clival DAVFs. Particular attention was given to safety and efficacy of transarterial embolization using liquid embolic agents. METHODS: We reviewed the clinical and radiological data of 10 patients with spontaneous clival DAVFs who were treated endovascularly at the University of California at Los Angeles Medical Center between 1992 and 2006. RESULTS: Nine patients presented with ocular symptoms and one patient experienced pulsatile tinnitus. Cerebral angiograms showed that these clival DAVFs were supplied by multiple branches of the internal and external carotid arteries. The patterns of venous drainage were from the clival veins to the cavernous sinus and superior ophthalmic vein in nine patients and to the inferior petrosal sinus in two patients. Six clival DAVFs were embolized transarterially through the clival branches of the ascending pharyngeal artery. Onyx 18 (Micro Therapeutics Inc., Irvine, CA) was used in three patients and n-butyl cyanoacrylate was used in three patients. Immediate complete angiographic obliteration was achieved in three patients. All six patients experienced an angiographic and clinical cure without any complications at 3 months. Two patients were incompletely treated using particles and coils for the relief of the symptoms. Two other patients were completely treated after the recipient clival venous structures were occluded transvenously with coils. CONCLUSION: Clival DAVFs can be misdiagnosed as dural cavernous sinus fistulae. The best treatment is transarterial embolization of the dural feeders using liquid embolic agents. Transvenous occlusion of the cavernous sinus is unnecessary in most cases. PMID- 18382319 TI - Intracranial vascular lesions associated with small epidural hematomas. AB - OBJECTIVE: We studied the angiographic findings in patients with small epidural hematomas and cranial fractures crossing over the trajectory of the middle meningeal artery and its branches. Additionally, the occurrence of traumatic vascular lesions and their clinical relevance and treatment are discussed. METHODS: A consecutive analysis was performed for 24 patients who harbored small epidural hematomas in middle meningeal artery topography associated with cranial fractures. Computed tomographic scans and plain x-ray studies were used to diagnose linear cranial fractures. Patients with large epidural hematomas or associated traumatic lesions were excluded from the study. Selective ipsilateral external carotid angiograms were obtained, and an endovascular procedure was performed if any vascular injury was evidenced. RESULTS: In all patients with cranial fractures crossing over the middle meningeal artery and its branches, some kind of vascular lesion was seen. Two types of findings were noted: active extravasation of the contrast medium (71%) and pseudoaneurysms (29%). Early filling of diploic vessels was found in 8.3% of fractures concomitantly with active extravasation. Embolization was performed in all patients. No additional enlargement of the epidural hematoma was observed, and the postoperative period was uneventful. CONCLUSION: This study shows that pseudoaneurysms and active extravasation of contrast are common findings in this subset of patients. Although the natural history of these lesions is still poorly understood, additional investigation with ipsilateral external carotid angiography may be recommended, considering the potentially catastrophic consequences of late rupture. PMID- 18382320 TI - Statin use was not associated with less vasospasm or improved outcome after subarachnoid hemorrhage. AB - OBJECTIVE: The development of delayed ischemia caused by cerebral vasospasm remains a common cause of morbidity and mortality after aneurysmal subarachnoid hemorrhage. Preliminary studies suggest that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) may decrease the risk of vasospasm, but additional study is required. METHODS: Beginning in May 2006, our treatment protocol for patients presenting with subarachnoid hemorrhage was altered to routinely include the use of 80 mg of simvastatin per day for 14 days. Before this time, only patients with other indications for statins were treated. The charts of 203 consecutive patients over a period of 27 months were retrospectively reviewed, and 150 patients were included in the analysis, of whom 71 patients received statins. These patients were compared with 79 untreated patients to determine whether or not the use of statins was associated with a reduction in the occurrence of vasospasm, delayed infarction, or poor outcome (death, vegetative state, or severe disability). RESULTS: Patients who were treated with statins and those who were not had similar baseline characteristics, although more patients in the former group were managed with endovascular coil embolization. There were no statistically significant differences in the proportion of patients developing at least moderate radiographic vasospasm (41% with statins versus 42% without, P = 0.91), symptomatic vasospasm (32% with statins versus 25% without, P = 0.34), delayed infarction (23% with statins versus 28% without, P = 0.46), or poor outcome (39% with statins versus 35% without, P = 0.61). After adjustment for differences in baseline characteristics, including the method of aneurysm treatment, statins were still not significantly protective. CONCLUSION: The addition of statins to standard care was not associated with any reduction in the development of vasospasm or improvement in outcomes after aneurysmal subarachnoid hemorrhage. If there is a benefit to statin use, it may be smaller than suggested by previous studies. However, further randomized controlled trials are awaited. PMID- 18382321 TI - Infectious complications of temporary spinal catheter insertion for diagnosis of adult hydrocephalus and idiopathic intracranial hypertension. AB - OBJECTIVE: Spinal catheters are often inserted for treatment of cerebrospinal fluid leaks; however, they have recently been recommended for elective cerebrospinal fluid drainage to identify patients with possible normal pressure hydrocephalus who are most likely to respond to shunt surgery. The rate of spinal catheter-associated meningitis with elective spinal catheter insertion is unknown. The objective was to determine the rate of infection and risk factors associated with elective spinal catheter insertion for evaluation of hydrocephalus and idiopathic intracranial hypertension (IIH). METHODS: We retrospectively analyzed clinical and microbiological data and cerebrospinal fluid results of patients admitted during 60 consecutive months who had elective spinal catheter insertion for evaluation of normal pressure hydrocephalus or IIH. RESULTS: A total of 461 spinal catheters were inserted in 454 patients, including 419 (90.9%) for treatment of hydrocephalus and 42 (9.1%) for IIH. The infection rate was 3.3% (15 out of 461 patients) for the entire cohort, 3.6% (15 out of 419 patients) for the hydrocephalus cohort, and 0% for the IIH cohort. There was one death (0.2%) in the hydrocephalus cohort. The infection rate was reduced and sustained at 1.8% for 225 catheters after the topical antiseptic was changed to chlorhexidine (two-sided Fisher's exact test; P = 0.114). CONCLUSION: Although infection is the most serious complication of spinal catheter insertion for evaluation of hydrocephalus or IIH, the infection rate can be maintained below 2% with use of chlorhexidine topical antiseptic application, single-dose preprocedural antibiotic administration, and clinical surveillance of the patient. The benefit of cerebrospinal fluid drainage via spinal catheter for normal pressure hydrocephalus outweighs the risks associated with the procedure. PMID- 18382322 TI - Laser-assisted endoscopic third ventriculostomy: long-term results in a series of 202 patients. AB - OBJECTIVE: Endoscopic third ventriculostomy is a well-known surgical option in the treatment of noncommunicating hydrocephalus. We studied complications and long-term success in 202 patients to demonstrate the safety and efficacy of laser assisted endoscopic third ventriculostomy (LA-ETV) using a unique "black" fiber tip/diode laser combination for controlled tissue ablation. METHODS: We studied 213 LA-ETVs, which were performed in 202 patients. Patients' ages ranged from 2 days to 83 years (mean age, 27 yr). The mean follow-up period for all patients was 2.7 years (range, 2 d to 12 yr). Hydrocephalus was caused by aqueductal stenosis in 65 patients, tumors in 67 patients, hemorrhages in 24 patients, myelomeningoceles in 20 patients, cysts in 15 patients, and other causes in 11 patients. The long-term effectiveness of LA-ETV was studied with Kaplan-Meier analysis. RESULTS: Technically successful LA-ETVs were accomplished in 196 of the 202 patients (97%). The overall success rate for a functional LA-ETV was 68% at the 2-year follow-up evaluation. LA-ETV was more effective in patients aged 1 year and older (70% success rate) than in younger patients (59% success rate). Success rates were greater in patients with aqueductal stenosis or tumors as compared to other etiologies. Complications occurred in 22 procedures (10.3%). Only one patient (0.5%) experienced a major complication. No surgical mortalities or laser-related complications occurred. CONCLUSION: This study demonstrates that LA-ETV is a safe and effective procedure that is comparable to other techniques for ETV. LA-ETV is most effective in patients aged 1 year and older and in patients with aqueductal stenosis and tumors, with a low major complication rate. PMID- 18382323 TI - A multi-institutional, 5-year analysis of initial and multiple ventricular shunt revisions in children. AB - OBJECTIVE: To evaluate risk factors and predictors of cerebrospinal ventricular shunt revisions in children. METHODS: A retrospective, longitudinal cohort of 1307 children ages 0 to 18 years undergoing initial ventricular shunt placement in the year 2000, with follow-up through 2005, from 32 freestanding children's hospitals within the Pediatric Health Information Systems database was studied. Rates of ventricular shunt revision were compared with patient demographic, clinical, and hospital characteristics with use of bivariate and multivariate regression accounting for hospital clustering. RESULTS: Thirty-seven percent of children required at least one shunt revision within 5 years of initial shunt placement; 20% of children required two or more revisions. Institutional rates of first shunt revision ranged from 20 to 70% of initial shunts placed among the 32 hospitals in the cohort. Hospitals where one to 20 initial shunt placements per year experienced the highest initial shunt revision rate (42%). Hospitals performing over 83 initial shunt placements per year experienced the lowest revision rate (22%). We found that children undergoing shunt placement in the Midwest were more likely to experience multiple shunt revisions (odds ratio, 1.25; 95% confidence interval, 1.06-1.47) after controlling for hospital volume, shunt type, age, and diagnosis associated with initial shunt placement. CONCLUSION: Higher hospital volume of initial shunt placement was associated with lower revision rates. Substantial hospital variation in the rates of ventricular shunt revision exists among children's hospitals. Future prospective studies are needed to examine the reasons for the variability in shunt revision rates among hospitals, including differences in specific processes of care. PMID- 18382324 TI - Cervical spondylotic myelopathy: complications and outcomes after spinal fusion. AB - OBJECTIVE: There is little information about in-hospital complication rates, adverse outcomes, and mortality after spinal fusion for cervical spondylotic myelopathy (CSM). The aim of this study was to report inpatient mortality, complications, and outcomes on a national level. METHODS: We used the National Inpatient Sample to identify 58,115 admissions of patients with CSM who underwent spinal fusion in the United States from 1993 to 2002. Multivariate analysis was performed to analyze the effects of patient and hospital characteristics on outcomes such as mortality, complications, discharge disposition, and length of stay. RESULTS: A total of 58,115 patients with CSM underwent spinal fusion with an average mortality rate of 0.6%, a complication rate of 13.4%, and a mean length of stay of 4 days. Pulmonary (3.6%) and postoperative hemorrhages or hematomas (2.3%) were the most common complications reported. One postoperative complication led to a 4-day increase in mean length of stay, increased the mortality rate 20-fold, and added more than $10,000 to hospital charges. Multivariate analysis identified age, comorbidity, and admission type as the main predictors of mortality, complication rate, and adverse outcome. Patients aged > or =85 or 65 to 84 years had respective 44- and 14-fold increases in mortality, compared with patients in the 18- to 44-year age group. Patients older than 84 years had a 40-fold increase in adverse outcomes and a 5-fold likelihood of medical complications. Patients with three or more comorbidities had an increased risk of medical complications (odds ratio [OR], 1.98), adverse discharge (OR, 2.17), and in-hospital mortality (OR, 2.36). Elective admissions were associated with much lower rates of mortality (OR, 0.28), complication (OR, 0.68), and adverse outcome (OR, 0.26). Complications were greater for posterior fusion (16.4%) versus anterior fusion (11.9%) procedures. Anterior fusions were associated with a greater incidence of dysphagia (3%) and hoarseness (0.21%). Cervical spondylosis patients who presented without myelopathy had a much lower incidence of complications (6.3%). CONCLUSIONS: We provide a national estimate of inpatient complications and outcomes after spinal fusion for CSM patients in the United States. We demonstrate the impacts of age, complications, and medical comorbidities on the outcome of surgery for patients with this common disorder. We provide complication rates stratified by age and medical comorbidities for elderly patients who present with CSM who need spinal fusion. PMID- 18382325 TI - Surgical management of anterior cranial base fractures with cerebrospinal fluid fistulae: a single-institution experience. AB - OBJECTIVE: The management of cerebrospinal fluid (CSF) fistulae after anterior cranial base fracture remains a surgical challenge. We reviewed our results in the repair of CSF fistulae complicating multiple anterior cranial base fractures via a combined intracranial extradural and intradural approach and describe a treatment algorithm derived from this experience. METHODS: We retrospectively reviewed the files of 209 patients with an anterior cranial base fracture complicated by a CSF fistula who were admitted between 1980 and 2003 to Liege State University Hospital. Among those patients, 109 had a persistent CSF leak or radiological signs of an unhealed dural tear. All underwent the same surgical procedure, with combined extradural and intradural closure of the dural tear. RESULTS: Of the 109 patients, 98 patients (90%) were cured after the first operation. Persistent postoperative CSF rhinorrhea occurred in 11 patients (10%), necessitating an early complementary surgery via a transsphenoidal approach (7 patients) or a second-look intracranial approach (4 patients). No postoperative neurological deterioration attributable to increasing frontocerebral edema occurred. During the mean follow-up period of 36 months, recurrence of CSF fistula was observed in five patients and required an additional surgical repair procedure. CONCLUSION: The closure of CSF fistulae after an anterior cranial base fracture via a combined intracranial extradural and intradural approach, which allows the visualization and repair of the entire anterior base, is safe and effective. It is essentially indicated for patients with extensive bone defects in the cranial base, multiple fractures of the ethmoid bone and the posterior wall of the frontal sinus, cranial nerve involvement, associated lesions necessitating surgery such as intracranial hematomas, and post-traumatic intracranial infection. Rhinorrhea caused by a precisely located small tear may be treated with endoscopy. PMID- 18382326 TI - Ballistics for the neurosurgeon. AB - Craniocerebral injuries from ballistic projectiles are qualitatively different from injuries in unconfined soft tissue with similar impact. Penetrating and nonpenetrating ballistic injuries are influenced not only by the physical properties of the projectile, but also by its ballistics. Ballistics provides information on the motion of projectiles while in the gun barrel, the trajectory of the projectile in air, and the behavior of the projectile on reaching its target. This basic knowledge can be applied to better understand the ultimate craniocerebral consequences of ballistic head injuries. PMID- 18382327 TI - Clinical problem solving: seize the day. PMID- 18382328 TI - A new multipurpose ventriculoscope. AB - This article describes a new multipurpose ventriculoscope that combines the advantages of several neuroendoscopic systems that are already available and provides some new features. This system can be used for all intracranial endoscopic procedures, such as endoscopic third ventriculostomy, septum fenestration, foraminoplasty, aqueductoplasty, stenting, cyst fenestrations, intraventricular tumor biopsy or resection, or intraventricular cyst or clot removal. The ventriculoscope can be used in both adults and children. Because of its diameter, it is not designed for the endoscopic treatment of newborns. PMID- 18382329 TI - Enhanced tumor growth elicited by L-type amino acid transporter 1 in human malignant glioma cells. AB - OBJECTIVE: To study the expression and function of L-type amino acid transporter 1 (LAT1), a major catalytic subunit of system L that is responsible for the transport of large neutral amino acids, including most essential amino acids, in concert with the covalently bound 4F2 heavy chain, and is implicated in tumorigenesis. METHODS: Human glioma cell lines and tumor specimens were analyzed for LAT1 expression using Western blotting and reverse transcription polymerase chain reaction analysis. The rate of neutral amino acid uptake was measured using L-[C]leucine. The proliferation and apoptosis rates were analyzed by 3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide and terminal deoxynucleotidyl transferase-mediated nick end-labeling assays, respectively, on inhibition of system L by 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid. The effects on proliferation and tumor growth caused by exogenously overexpressed LAT1 were similarly analyzed. RESULTS: LAT1 was expressed in most human high grade gliomas and glioma cell lines at various levels, with more ubiquitous expression of 4F2 heavy chain. Glioma cells with high LAT1 expression exhibited a marked increase in the uptake rate of L-[C]leucine. 2-Aminobicyclo-(2,2,1) heptane-2-carboxylic acid treatment not only suppressed deoxyribonucleic acid synthesis in association with the up-regulation of the cyclin-dependent kinase inhibitor p21 but also enhanced apoptosis with caspase activation, thereby exerting both cytostatic and cytocidal effects in glioma cells with high LAT1 expression levels. Furthermore, overexpression of LAT1 in glioma cells with low endogenous LAT1 expression significantly enhanced the rates of tumor cell growth in athymic mice. CONCLUSION: LAT1, the major transporter of system L, is frequently expressed at higher levels in high-grade gliomas than in low-grade gliomas and brain tissues, and it may play an important role in enhancing the rates of tumor cell proliferation and growth in vivo. PMID- 18382330 TI - Identification of A2B5+CD133- tumor-initiating cells in adult human gliomas. AB - OBJECTIVE: Several studies have shown that human gliomas contain a small population of cells with stem cell-like features. It has been proposed that these "cancer stem cells" may be uniquely responsible for glioma formation and recurrence. However, human gliomas also contain an abundance of cells that closely resemble more differentiated glial progenitors. Animal model studies have shown that these cells also possess the capacity to form malignant gliomas. METHODS: To investigate the contributions of stem-like and progenitor-like cells in human gliomas, we used flow cytometry to characterize the expression of a cancer stem cell marker (CD133) and a glial progenitor marker (A2B5) in 25 tumors. We found that human gliomas consistently express A2B5 in a large percentage of cells (61.7 +/- 3.8%, standard error of the mean). In contrast, CD133 expression was less abundant and less consistent (14.8 +/- 3.6%, standard error of the mean), with several glioblastomas containing very few or no detectable CD133+ cells. When present, the CD133+ population was almost entirely contained within the A2B5+ population. Thus, most gliomas could be divided into three distinct populations on the basis of these markers (A2B5+CD133+, A2B5+CD133 , and A2B5-CD133-). To test the tumorigenic potential of these populations, we separated cells from six tumors by fluorescence-activated cell sorting and reinjected them into nude rats. RESULTS: We found that the capacity for these different populations to form tumors varied depending on the human tumor specimen from which they were isolated. Of the six human gliomas tested, four contained A2B5+/CD133- cells that formed tumors when transplanted into nude rats, three contained A2B5+/CD133+ cells that formed tumors, and only one glioma contained A2B5-/CD133- cells with the capacity to form tumors. CONCLUSION: Together, these results demonstrate that human gliomas contain multiple populations of cells with the capacity to form tumors and specifically identify a population of tumorigenic A2B5+ cells that are phenotypically distinct from CD133+ cells. PMID- 18382331 TI - The status quo of neurosurgery in China. AB - With the application of great effort, much progress has been made to date in each specialty of neurosurgery in mainland China. In this article, we briefly review the present status of neurosurgery in China. The components and function of the Chinese Neurosurgical Society, the national organization for neurosurgery in China, are discussed. Neurosurgeons' acceptance of the concept of minimally invasive procedures has marked the start of an era of minimally invasive neurosurgery in China. Progress is evident in clinics, basic research, infrastructure, resident training, and multidisciplinary collaboration. Some weaknesses that need improvement are also mentioned. The current program offers a good basic foundation for development to meet future demands. PMID- 18382332 TI - "Man, controller of the universe" by Diego Rivera (1886-1957). PMID- 18382333 TI - Ophthalmic manifestations of an undifferentiated sinonasal carcinoma. AB - INTRODUCTION: Sinonasal undifferentiated carcinoma (SNUC) are uncommon, highly aggressive neoplasms arising in the nasal cavity and paranasal sinuses. Unlike the majority of sinonasal carcinomas, SNUC possess an uncertain histological origin and pleomorphic composition. Their heterogeneous composition prohibits their distinction from other more commonly encountered nasal tumors on the basis of imaging alone. Patients afflicted with SNUC often complain of symptoms of chronic, progressive sinus congestion, and epistaxis. Imaging studies of SNUC often illustrate rapidly enlarging masses involving multiple sinonasal sites with frequent evidence of extension beyond the sinonasal tract. Orbital extension is a frequent finding encountered during radiographic investigations. CASE REPORT: A 28-year-old white man presented complaining of diplopia in superior gaze, pain on eye movement, and gaze-evoked amaurosis which progressed during a 2-day period. Funduscopic examination revealed choroidal folds in the papillomacular region OS. Magnetic resonance imaging (MRI) revealed a 3 cm x 3.5 cm x 4 cm mass emanating from the left ethmoid sinus with bony destruction of the left orbital floor, nasal wall of the left and right orbits, and the cribriform plate. A transcutaneous aspiration fine-needle biopsy confirmed the presence of a SNUC. After a 7-wk course of combined chemoradiotherapy marked regression in the mass was attained with resolution of the ophthalmic symptoms. CONCLUSION: SNUC is an aggressive malignancy that is often discovered in its advanced stages after having afflicted serious insult on the nasal and sinus cavities. Ocular involvement is feasible owing to the thin composition of the orbital walls and the fast growing nature of the neoplasm. PMID- 18382334 TI - Factors affecting corneal and conjunctival sensitivity measurement. AB - PURPOSE: Measurement of sensitivity provides important clues about sensation on the ocular surface. This study aimed to evaluate whether measurements of threshold of sensation to an air stimulus are affected by corneal/conjunctival location, gender, age, time of day, ambient temperature or humidity. METHODS: A retrospective analysis is reported of ocular surface threshold measurements made by one examiner using the CRCERT-Belmonte esthesiometer. Multiple corneal measurements for 49 normal subjects (24M:25F) and conjunctival measurements for 33 subjects (16M:17F) were included in the analysis. Threshold was measured at the corneal apex and at the inferior conjunctiva 2 mm from the limbus. Measurements were made between 9 AM and 6 PM, at ambient temperature 20 to 26 degrees C and humidity 52 to 87%. Mixed model analysis of variance, paired-t-test and Pearson's correlation were used to examine effects of various factors on threshold. RESULTS: Mean group corneal threshold was 76.2+/-26.8 mL/min and conjunctival threshold 123.7+/-49.1 mL/min (n=33, p<0.001). Corneal and conjunctival threshold were well correlated (r=0.66, p<0.001). Thresholds were significantly higher for male than female subjects at both the cornea (M 82.2+/ 23.5 mL/min, F 67.6+/-24.1 mL/min, p=0.04) and conjunctiva (M 144.1+/-40.7 mL/min, F 105.8+/-50.2 mL/min, p=0.02). A significant reduction in corneal threshold with age was apparent for females (n=25, r= -0.49 p=0.01) but not males. A similar effect on conjunctival sensitivity was not shown. No effect of time of day, ambient humidity or temperature was found on threshold at either site. CONCLUSIONS: Corneal and conjunctival sensitivity were found to be associated. Corneal and conjunctival sensitivity is higher in female subjects, who also show an age-related increase in corneal sensitivity. No change in sensitivity of either tissue is apparent within normal levels of ambient temperature or humidity or over the course of a working day. PMID- 18382335 TI - Retinal vessel analysis reproducibility in assessing cardiovascular disease. AB - PURPOSE: To investigate the clinical accuracy and determine the reproducibility of static, semiautomated retinal vessel analysis to supplement established vascular risk factors. METHODS: Manual blood pressure measurements and calibrated retinal photographs were obtained, after informed consent, on subjects without any eye disease aged >50 years. A total of 48 subjects without systemic hypertension or any other vascular disease and 54 subjects with confirmed hypertension were enrolled. Analysis was performed on retinal photographs taken by a retinal thickness analyzer (Talia Technology, Israel). The arteriovenous ratio (AVR) was calculated by a semiautomated vessel tracking VSL software (Talia Technology). Reproducibility was determined for software tracking, intra-, and intergrader selection as well as intra- and intervisit for 20 subjects. The effects of image quality degradation and decentration were investigated. RESULTS: Validation showed an excellent agreement between semiautomated software and manual vessel measurements. In the 102 subjects analyzed, retinal AVR was only correlated with established systemic hypertension (p=0.01) and gender (p=0.01). There was no effect of age on AVR. Other risk factors such as diabetes, smoking, body mass index, and current blood pressure showed some trends on multifactorial analysis. When limiting the number of vessels selected, software tracking induced no variability. The mean standard deviation for AVR was 0.02 for intra- as well as intergrader and 0.01 for inter- and intravisit effects. Image decentration only increased variability and the algorithm was robust against reducing image resolution and noise. Improper image focus, however, caused incorrect measurements of AVR. CONCLUSIONS: Analysis of AVR can be performed reproducibly on routine retina photographs. Retinal AVR appears to be a relatively independent risk factor to assess systemic cerebrovascular disease. PMID- 18382336 TI - Whither goes evidence-based optometry? PMID- 18382338 TI - Dynamic simulation of the effect of soft toric contact lenses movement on retinal image quality. AB - PURPOSE: To report the development of a tool designed to dynamically simulate the effect of soft toric contact lens movement on retinal image quality, initial findings on three eyes, and the next steps to be taken to improve the utility of the tool. METHODS: Three eyes of two subjects wearing soft toric contact lenses were cyclopleged with 1% cyclopentolate and 2.5% phenylephrine. Four hundred wavefront aberration measurements over a 5-mm pupil were recorded during soft contact lens wear at 30 Hz using a complete ophthalmic analysis system aberrometer. Each wavefront error measurement was input into Visual Optics Laboratory (version 7.15, Sarver and Associates, Inc.) to generate a retinal simulation of a high contrast log MAR visual acuity chart. The individual simulations were combined into a single dynamic movie using a custom MatLab PsychToolbox program. Visual acuity was measured for each eye reading the movie with best cycloplegic spectacle correction through a 3-mm artificial pupil to minimize the influence of the eyes' uncorrected aberrations. Comparison of the simulated acuity was made to values recorded while the subject read unaberrated charts with contact lenses through a 5-mm artificial pupil. RESULTS: For one study eye, average acuity was the same as the natural contact lens viewing condition. For the other two study eyes visual acuity of the best simulation was more than one line worse than natural viewing conditions. CONCLUSIONS: Dynamic simulation of retinal image quality, although not yet perfect, is a promising technique for visually illustrating the optical effects on image quality because of the movements of alignment-sensitive corrections. PMID- 18382339 TI - Hydrogel contact lens binding induced by contact lens rewetting drops. AB - PURPOSE: To investigate alterations in prelens tear film (PLTF), contact lens (CL), and postlens tear film (PoLTF) thicknesses and contact lens movement associated with boric acid-containing rewetting drops. METHODS: This was a single center, randomized, double-masked clinical trial. Following etafilcon A lens application, baseline lens movement and interferometric measures (PLTF, CL, PoLTF thicknesses) were captured. A randomly assigned drop (either a low or high boric acid-containing drops) was applied to both eyes, followed immediately by lens movement and interferometric measures. After a 2-h washout period (spectacle wear), subjects returned for testing with the alternative drop. Statistical analyses addressed pre- to postdrop changes in lens movement and PLTF, CL, and PoLTF thickness. RESULTS: The presence of a drop alone lead to substantial decreases in contact lens movement (general reduction of 0.45 mm), but the reduction was not related to drop type. The presence of a drop led to an increase in PLTF thickness (approximately 0.14 microm), again regardless of the drop type. Both the general presence of a rewetting drop and drop type lead to an increase in CL center thickness (lower boric acid drop = 0.31 microm and higher boric acid drop = 1.13 microm). Both the general presence of a rewetting drop and the drop type lead to a significant decrease in PoLTF thickness, whereby the lower boric acid drop lead to a decrease of 0.28 mum and the higher boric acid drop lead to a decrease of 0.73 microm. CONCLUSIONS: Boric acid-containing rewetting drops were shown to be associated with a significant reduction in lens movement with a concurrent increase in lens center thickness and reduction in PoLTF thickness. The higher boric acid-containing drop was associated with more substantial increases in lens center thickness and decreases in PoLTF thickness. PMID- 18382340 TI - Feasibility of using placebo vision therapy in a multicenter clinical trial. AB - PURPOSE: The Convergence Insufficiency Treatment Trial (CITT) Investigator Group conducted a preliminary study assessing the effectiveness of home-based push-up therapy and office-based vision therapy/orthoptics for the treatment of convergence insufficiency (CI). The CITT group developed a placebo therapy program that was designed to simulate real vision therapy/orthoptics. The purpose of this paper is to evaluate the effectiveness of this placebo therapy program in maintaining masking of subjects randomized to the office-based treatment arms (real or placebo). METHODS: Subjects (ages 9 to 30 years) were enrolled, stratified into two groups by age, and then randomly assigned to one of three treatment groups: pencil push-up therapy, office-based vision therapy/orthoptics, or office-based placebo vision therapy/orthoptics. At the end of treatment, subjects in the two office-based therapy groups (placebo and real) were asked: (1) which treatment do you think you received? and (2) how sure are you about your answer? RESULTS: Ninety-five percent of subjects assigned to real therapy and 83% assigned to placebo therapy thought they were in the real therapy group. Of the subjects who thought they received real therapy, 90% assigned to real therapy and 89% assigned to placebo therapy were "somewhat sure," "pretty sure," or "very sure" of their answer. Those assigned to real therapy had more responses in the "very sure" category. CONCLUSION: The CITT placebo therapy program was effective in maintaining subject masking in this multicenter clinical trial. PMID- 18382341 TI - The evidentiary significance of case reports: eye rubbing and keratoconus. AB - PURPOSE: Individual case reports (observational or interventional) are thought to have a very low ranking for evidentiary significance. This clinical report examines the relationship between the evidentiary significance of case reports and their presentation. METHODS: An observational report of unilateral keratoconus (KC), that developed in association with a corresponding unilateral chronic habit of abnormal rubbing, is examined to identify features that can raise the level of evidentiary significance. RESULTS: A feature of this type of case is the opportunity to compare the eye with KC to the unaffected eye, for which there was an unremarkable rubbing history. The unaffected eye serves as a retrospective unintentional control for the affected eye. Detailed assessment to establish the normality of the apparently normal eye is crucial to the differential diagnosis between a markedly asymmetric presentation and a true unilateral presentation. Any history of systemic diseases that may be associated with the presenting disease can be important. Longitudinal follow-up, especially if postonset data are supplemented with preonset data, can contribute to the evidentiary base by allowing for comparison with the natural history of the relevant disease. Intervention to remove a suspected causal factor, when combined with appropriate follow-up, can provide additional evidence. CONCLUSIONS: Depending on their presentation, some types of case report can have a significant evidence-based function in assessing causal hypotheses and, accordingly, provide a higher evidentiary ranking than generally assigned to case reports. The potential for a higher evidentiary ranking may be more important when randomized clinical trials are contraindicated for ethical reasons, or there are challenging problems with study design. Wider agreement regarding the quantitative and qualitative criteria for diagnosing the earliest stages of KC would aid the assessment of the evidentiary significance of case reports of this disease. PMID- 18382342 TI - Retinal image location of hand, fingers, and objects during manual tasks. AB - PURPOSE: A new method was developed using the scanning laser ophthalmoscope (SLO) to investigate the effects of central visual loss on eye-hand coordination in manual tasks. Using the SLO, the retinal positions of the hand, fingers, and objects are imaged and recorded while a subject performs a manual task. METHOD: A video camera images the subject's hand and objects to be manipulated in the SLO laser-beam raster, producing a video image of a subject's hand, fingers, and objects on the subject's retina while the objects are manipulated. A subject with bilateral central scotomas and an age-matched control subject with normal vision traced an ellipse with the index finger, tapped four disks in sequence, and carried out a pattern duplication task with pegs. Retinal positions of the fovea or preferred retinal locus (PRL), fingers, and objects were measured from digitized SLO images. RESULTS: In all tasks, the fovea or PRL was directed to an object or position before the fingers arrived. This lead time was much greater for the scotoma subject than the control subject ( approximately 1400 vs. approximately 400 ms, respectively). The scotoma subject was much less accurate in placing the PRL and fingers on objects and required substantially more time for task completion than the control subject. CONCLUSIONS: The coordination of foveal fixation and finger placement found with the SLO method was similar to that found by others using eyetracking techniques with visually normal subjects. The presence of a central scotoma and use of a PRL caused marked deterioration in the quality of this coordination. Unlike eyetracking methods, the SLO technique does not require calibration because the positions of the fingers and objects are directly observable on the retina. This method could be useful in studying eye hand coordination of individuals with scotomas that affect foveal vision. PMID- 18382344 TI - Preoperative assessment of corneal and refractive stability in soft contact lens wearing photorefractive candidates. PMID- 18382345 TI - Changes in lens dimensions and refractive index with age and accommodation. PMID- 18382347 TI - Assessing the impact of tissue devitalization time on genome-wide gene expression analysis in ovarian tumor samples. AB - The utilization of genome-wide gene expression microarray technology in tumor stratification has proven a powerful tool to identify gene expression signatures associated with cancer prognosis and is currently under evaluation in clinical laboratories. Standardized protocols, including tumor tissue postoperatively handling guidelines are yet to be defined. We aimed at assessing a systematic effect of devitalization in ovarian tumors' gene expression profiling, using high density oligonucleotide microarrays, under a standardized protocol following strict quality control criteria. Residual tissue from the surgical pathology specimen was divided into 5 samples. Half of each was immediately snap frozen in liquid nitrogen. The remaining halves were kept at room temperature for 0, 15, 30, 60, and 120 minutes, at which time the tissue was snap frozen in liquid nitrogen, and stored at -80 degrees C until RNA extraction. The entire process from RNA extraction through feature intensity distribution was rigorously monitored for quality. Identification of altered gene expression among each pair of snap frozen and devitalized samples per ovarian tumor specimen was assessed by using the Significance score (S-score) method. We identified only 4 probe sets that seemed to correlate with devitalization time in one of the ovarian tumor specimens, suggesting that they are not likely to have an impact on gene expression profiling tumor stratification. Our study suggests that with proper sample handling and rigorous quality control procedures for RNA extraction and microarray analysis, tumor classification based on global gene expression data will not be adversely affected if devitalization times are kept within a 120 minute window. PMID- 18382348 TI - DNA-based polymerase chain reaction for detecting FUS-CREB3L2 in low-grade fibromyxoid sarcoma using formalin-fixed, paraffin-embedded tissue specimens. AB - Detection of the tumor-specific fusion gene, FUS-CREB3L2, is useful in the diagnosis of low-grade fibromyxoid sarcoma (LGFMS), especially by reverse transcription (RT)-polymerase chain reaction (PCR) using formalin-fixed, paraffin embedded (FFPE) tumor tissues. Because FUS and CREB3L2 gene segments with their break points within exons are fused without interposed introns in many LGFMSs, DNA-based PCR may also be applicable to detect the FUS-CREB3L2 fusion gene using FFPE specimens. In this study, DNA and RNA were extracted from FFPE specimens of 16 and 19 LGFMSs, respectively, and PCR and RT-PCR were performed using primers that specifically amplify most of the junctional regions of the FUS-CREB3L2 fusion gene. RT-PCR analysis revealed FUS-CREB3L2 fusion transcripts in 16 of the 19 (84%) LGFMSs. In 14 informative examples, including 11 with detectable fusion transcripts by RT-PCR, DNA-based PCR amplified the FUS-CREB3L2 fusion gene in 9 (64%) examples. Nucleotide sequence analysis confirmed that PCR products were identical to the respective RT-PCR products in 8 cases. Although the sensitivity was not as high as RT-PCR, DNA-based PCR can be used as an alternative molecular approach for detecting the FUS-CREB3L2 fusion gene. PMID- 18382350 TI - Utility of chromogenic in situ hybridization (CISH) for detection of EGFR amplification in glioblastoma: comparison with fluorescence in situ hybridization (FISH). AB - In this study, we test the reliability of chromogenic in situ hybridization (CISH) for the detection of epidermal growth factor receptor (EGFR) gene amplification in glioblastoma. Earlier reports have described EGFR CISH in glioblastoma multiforme, but a comparison of CISH with a "gold standard" testing method, such as fluorescence in situ hybridization (FISH), has not been described. Therapies targeting the EGFR-signaling pathway might increase the importance of assessment of EGFR-amplification status. CISH is a potential alternative to FISH as a testing method. To test its reliability, EGFR amplification status by CISH was assessed in 89 cases of glioblastoma and compared with FISH results, and correlated with the protein expression using immunohistochemistry (IHC) for EGFR. FISH was scored as being EGFR-amplified in 47/89 tumors, CISH as being amplified in 43/89 tumors. The CISH and FISH results were in agreement in 83/89 cases (93%). Four glioblastomas were scored as being amplified by FISH, but not by CISH; whereas amplification was detected in 2 tumors by CISH that were not amplified using FISH. Forty-eight of the 89 cases were positive for EGFR expression by IHC. EGFR amplification was highly correlated with protein expression by IHC, as 40/48 (83%) EGFR IHC-positive cases were found to be EGFR-amplified. The high concordance of CISH and FISH for the assessment of EGFR gene-amplification status indicates that CISH is a viable alternative to FISH for the detection of EGFR gene amplification in glioblastoma. Detectable EGFR expression by IHC can occur in the absence of gene amplification, but is uncommon. PMID- 18382351 TI - Amplification of mRNA from laser-microdissected single or clustered cells in formalin-fixed and paraffin-embedded tissues for application in quantitative real time PCR. AB - The determination of marker genes and gene clusters involved in disease pathogenesis is increasingly contingent on high-throughput methods of gene expression profiling. However, the concurrently increasing application of mRNA from formalin-fixed and paraffin-embedded (FFPE) tissue archives, as well as cell type-specific approaches by laser-assisted microdissection, frequently results in very small and degraded quantities of RNA. Therefore, a successful amplification of cell-type-specific mRNA targets from FFPE tissues becomes more and more essential. To optimize the hitherto limited technical options, we applied 3 commercial amplification kits on FFPE single cells. We thereby determined the approach of target-specific cDNA amplification as being notably appropriate for subsequent real-time polymerase chain reaction, as a constant decrease of CT values by 14 polymerase chain reaction cycles could be demonstrated. PMID- 18382352 TI - Real time RT-PCR approach for the evaluation of ERBB2 overexpression in breast cancer archival samples: a comparative study with FISH, SISH, and immunohistochemistry. AB - We tested the reliability of real time reverse transcription polymerase chain reactions as an alternative method for the assessment of ERBB2 status in paraffin embedded tissues of 83 patients with breast cancer and 20 non-neoplastic controls. PCR was also compared with the immunohistochemical (IHC) HercepTest score and with fluorescence (FISH) and silver (SISH) in-situ hybridization, in 42 selected cases. ERBB2 mRNA was overexpressed in 26/83 (31%) breast cancer samples, using a cutoff calculated as the mean value of the controls plus 3 SD or with the receiver operating curve. The PCR test showed a 96% sensitivity and a 100% specificity when compared with FISH, with an area under the receiver operating curve of 98.4%. Overexpression of ERBB2 at PCR was also significantly correlated with amplification in FISH (P<0.001, Mann-Whitney test) and in SISH (P<0.001, Mann-Whitney test), and with the IHC HercepTest scores 2 or 3 (P<0.001, Spearman rank correlation). FISH, SISH, and IHC were also compared with each other. ERBB2 amplification in FISH significantly correlated with that in SISH (P=0.002, chi test with a concordance of the 87%), but not with IHC HercepTest scores (P=0.214, chi test). Real time PCR is a reliable and cost-effective method for the assessment of ERBB2 status in archival breast cancer samples, compared with FISH. Its introduction in routine diagnostic pathology practice is feasible even if it requires amendments to the current clinical oncology protocols. PMID- 18382353 TI - Correlation of Pap smear, cervical biopsy, and clinical follow-up with an HPV typing microarray system. AB - A human papillomavirus (HPV) microarray system allows the determination of HPV type in clinical samples. The purpose of this study was to determine the presence of HPV in liquid-based Pap smears with the MyGene MyHPV Chip Kit HPV genotyping microarray test (MyGene Assay), and to correlate this with the cytology and biopsy diagnoses, clinical follow-up, HPV Hybrid Capture data, and HPV sequence analyses. Four hundred and two Pap smears (93 ThinPrep, 309 SurePath) were available for study. Correlation of HPV DNA detection by the MyGene Assay with the Pap smear diagnosis showed a detection rate of 19/97 (19%) for normal Pap smears, 181/242 (74%) for atypical squamous cells of undetermined significance (ASCUS), and 61/63 (97%) for squamous intraepithelial lesions (SILs). Biopsy data on 248 women were available. HPV was noted by the MyGene Assay in the Pap smear in 98/100 (98%) of the cases, for which the corresponding biopsy had been diagnosed as SIL, compared with 103/148 (69%) of the cases for which the biopsy had been negative for SIL. Clinical follow-ups were available for 200 women with ASCUS Pap smears. A significant increase was observed in the rate of biopsy proven SILs in women with ASCUS Pap smears that were HPV-positive (63/66=95%) as compared with those that were HPV-negative (96/134=71%, P<0.05). The MyGene Assay and Hybrid Capture system gave equivalent results for all the categories studied, except for the presence of multiple infections, as determined by viral sequence analysis. Specifically, the Hybrid Capture system overestimated the presence of dual infection (low-risk and high-risk positive) by 48% and missed many cases of multiple infections, especially when 2 or more high-risk types were present. It is concluded that the MyGene Assay allows for the routine typing of HPV in liquid based Pap smears, and that the presence of HPV DNA in ASCUS Pap smears is strongly correlated with a biopsy-proven SIL. PMID- 18382354 TI - Keratin-positive gastrointestinal stromal tumor of the stomach mimicking gastric carcinoma: diagnosis confirmed by c-kit mutation analysis. AB - In routine practice, gastrointestinal stromal tumor (GIST) can usually be identified with relative ease on the basis of a rather simple immunohistochemical panel besides its characteristic morphology. Still, serious differential diagnostic problems may arise because of the heterogeneity of these tumors in both morphologic appearance and clinical behavior. In our case, we present a metastatic, ulcerative, hemorrhagic GIST with epithelioid appearance, which displayed diffuse pan cytokeratin (AE1/AE3) positivity beside CD117 expression. As carcinomas may also be CD117-positive, definitive diagnosis was confirmed by the detection of a hexanucleotide deletion in the exon 11 of c-kit. This case demonstrates that although gastric carcinoma more commonly ulcerates or causes hemorrhage than GIST, keratin-positive GIST should also be considered from a differential diagnostic point of view. In these cases, c-kit mutation analysis may be necessary. PMID- 18382355 TI - Sentinel lymph node molecular pathology in breast carcinoma. AB - OBJECTIVES: The prognosis of breast cancer patients depends on primary tumor resection and axillary lymph nodes examination. The purpose of this study was to analyze by molecular biology techniques the presence of mammaglobin A and B messenger RNA in breast sentinel lymph node (SLN) by reverse-transcription polymerase chain reaction (RT-PCR). METHODS: Sentinel lymph nodes from 50 patients with a diagnosis of breast cancer were prospectively studied between June 2004 and August 2006. Lymph nodes were all examined every 2 mm by intraoperative cytology. Hematoxylin-eosin (HE), immunohistochemistry (IHC) with cytokeratin (clone AE1-AE3, DAKO, dilution 1:100), and molecular biology techniques were used in all cases. RESULTS: Deferred study with routine techniques showed subcapsular metastasis in 3/50 cases. Out of 50 cases, 5 were detected with IHC, and 2 of them were negative for HE. Multiplex RT-PCR allowed the detection of 18/50 positive SLN, which included the 5 above-mentioned cases. The other SLN studied (32/50) showed no metastases with the methods herein implemented. CONCLUSIONS: The epidemiologic impact of incomplete SLN study has been observed, as the HE technique fails to identify all SLN with micrometastases. In our opinion, SLN should be studied with IHC and molecular biology techniques. The multiplex RT-PCR technique for A and B mammaglobin proves to be specific and sensitive. This study will serve to formulate hypotheses. Further research, including a larger population and a longer-term follow-up period, will be required to confirm these hypotheses. Should our findings be confirmed in the future, molecular biology determinations could modify patients' staging and treatment. PMID- 18382356 TI - Alveolar soft part sarcoma: a bimarker diagnostic strategy using TFE3 immunoassay and ASPL-TFE3 fusion transcripts in paraffin-embedded tumor tissues. AB - Alveolar soft part sarcoma (ASPS) is a malignancy with low incidence, but with poor prognosis if misdiagnosed. Immunohistochemical assay using TFE3 antibody has been shown to be a sensitive technique for ASPS diagnosis. A specific chromosomal translocation, t(X;17)(p11.2;q25), results in the ASPL-TFE3 fusion gene: it is detectable using reverse-transcription polymerase chain reaction (RT-PCR) in frozen tumor tissues of ASPS. However, the diagnostic usefulness of these markers has not been investigated in Chinese ASPS patients. Here, we report the first systematic study applying TFE3 immunoassay and ASPL-TFE3 fusion transcript detection to archival paraffin-embedded tissues in a large Chinese ASPS patient population. Sixteen patients had been diagnosed with ASPS (age, 3 to 58 y; 3 male patients and 13 female patients). Their tumors presented predominantly in the extremities (8/16), and were often located in the region of the orbit when affecting infants and children (3/16). Others had tumors in the chest wall, breast, and right pubis, respectively. One patient exhibited a tumor in the renal hilum, a location that had not been previously reported. Two patients had tumor metastases in the lung and the brain. ASPS tumors showed the best immunoreactivity to the TFE3 antibody (16/16). However, their immunoreactivity to other antibodies, including myoglobin (13/16), actin (10/16), desmin (2/16), and vimentin (2/16), was of various degrees. Positive staining was observed for the neural markers, NSE (9/16) and CgA (7/16), respectively. Using a strategy of RT PCR, followed by a nested PCR with a different primer set, we were able to detect the expression of the chimeric ASPL-TFE3 mRNA in 11 of the 16 ASPS tumors. Of these 11, 7 were type 1 ASPL-TFE3 and 4 were type 2 ASPL-TFE3, including the tumor located in the renal hilum. No expression of ASPL-TFE3 fusion transcripts was detectable in all 38 control tumors. Our results demonstrate that the "bimarker strategy," a combination of TFE3 immunostaining and ASPL-TFE3 chimeric transcript detection, might have sufficient sensitivity and specificity in diagnosing most of the ASPSs. As both diagnostic techniques can be applied to widely available archival paraffin-embedded tissues, the usefulness of the strategy is largely implicated in routine pathology laboratories. PMID- 18382357 TI - Primary mixed squamous carcinoma and osteosarcoma (carcinosarcomas) of the lung have a CGH mapping similar to primitive squamous carcinomas and osteosarcomas. AB - Carcinosarcomas are malignant tumors with a mixture of carcinomatous and differentiated sarcomatous elements. We investigate the morphology, immunohistochemistry, and comparative genomic hybridization analysis of 3 mixed squamous carcinoma and osteosarcoma of the lung. All patients were male and their ages were 72, 43, and 58 years. The sizes of the neoplasms were 7, 5, and 5 cm in maximum diameter, respectively. Two patients died of the disease 9 and 14 months after surgery; and one is alive 6 months later. By light microscopy, all cases had both squamous and osteosarcomatous structures. Immunohistochemistry was positive for AE3AE1, p63, 34 E12, CAM 5.2 (2/3 cases), CK-7 (2/3 cases), epithelial membrane antigen, E-cadherin, p53, and carcinogenic embryonic antigen in carcinomatous areas, and for vimentin and CD-68 in sarcomatous component. Areas of transition positive for both cytokeratins and vimentin were seen in all cases. A total of 55 copy number changes were detected with a median of 18 abnormalities per case: 48 gains, 6 losses, and 1 high-level amplification. Chromosome alterations in osteosarcomatous areas were similar to those found in lung metastatic osteosarcoma, comparable to those found in carcinomatous areas and to lung squamous carcinomas. Coincidences between carcinomatous areas and osteosarcomatous zones were found as gains in chromosomes 1q, 3q, 5p, 8q, and 12p. These findings provide arguments that favor a common origin for both types of cells, supported by the mixture of cells, the existence of undifferentiated cells positive to both cytokeratin and vimentin markers, and the CGH overlaps of chromosomal gains between carcinomatous and sarcomatous areas. PMID- 18382358 TI - Pyrosequencing analysis for detection of a BRAFV600E mutation in an FNAB specimen of thyroid nodules. AB - BACKGROUND: Fine-needle aspiration biopsy (FNAB) is the primary means of distinguishing benign from malignant and of guiding therapeutic intervention in thyroid nodules. However, 10% to 30% of cases with indeterminate cytology in FNAB need other diagnostic tools to refine diagnosis. OBJECTIVE: We compared the pyrosequencing method with the conventional direct DNA sequencing analysis and investigated the usefulness of preoperative BRAF mutation analysis as an adjunct diagnostic tool with routine FNAB. METHODS: A total of 103 surgically confirmed patients' FNA slides were recruited and DNA was extracted after atypical cells were scraped from the slides. BRAF mutation was analyzed by pyrosequencing and direct DNA sequencing. RESULTS: Sixty-three (77.8%) of 81 histopathologically diagnosed malignant nodules revealed positive BRAF mutation on pyrosequencing analysis. In detail, 63 (84.0%) of 75 papillary thyroid carcinoma (PTC) samples showed positive BRAF mutation, whereas 3 follicular thyroid carcinomas, 1 anaplastic carcinoma, 1 medullary thyroid carcinoma, and 1 metastatic lung carcinoma did not show BRAF mutation. None of 22 benign nodules had BRAF mutation in both pyrosequencing and direct DNA sequencing. Out of 27 thyroid nodules classified as 'indeterminate' on cytologic examination preoperatively, 21 (77.8%) cases turned out to be malignant: 18 PTCs (including 2 follicular variant types) and 3 follicular thyroid carcinomas. Among these, 13 (61.9%) classic PTCs had BRAF mutation. None of 6 benign nodules, including 3 follicular adenomas and 3 nodular hyperplasias, had BRAF mutation. Among 63 PTCs with positive BRAF mutation detected by pyrosequencing analysis, 3 cases did not show BRAF mutation by direct DNA sequencing. Although it was not statistically significant, pyrosequencing was superior to direct DNA sequencing in detecting the BRAF mutation of thyroid nodules (P=0.25). CONCLUSION: Detecting BRAF mutation by pyrosequencing is more sensitive, faster, and less expensive than direct DNA sequencing and is proposed as an adjunct diagnostic tool in evaluating thyroid nodules of indeterminate cytology. PMID- 18382359 TI - Mutations of the PIK3CA gene in diffuse large B cell lymphoma. AB - The PI3K/AKT pathway might be involved in the development of some certain diffuse large B-cell lymphoma (DLBCL) by as-yet unclear mechanisms. PIK3CA mutations in exons 9 and 20 were investigated in 76 primary human DLBCLs, 3 DLBCL cell lines (LY1, LY8, and LY10), and 9 related samples using polymerase chain reaction-based sequence analysis to assess the possible relevance of PIK3CA mutations in DLBCL to the PI3K/AKT pathway activation. AKT phosphorylation (pAKT) of 3 DLBCL cell lines and 76 primary DLBCL samples was also detected by Western blot and immunohistochemistry. All 3 cell lines showed high levels of pAKT, and 72.4% (55/76) of the DLBCLs expressed pAKT at various levels, indicating the activation of AKT. However, no mutation was found in exons 9 or 20 in PIK3CA in any of the 3 cell lines. Only 1 out of 76 primary DLBCLs (1.32%) harbored an exon 9 mutation, and no exon 20 mutation was detected. The case with mutations contained 3 mutation points. One was c.1634A>C resulting in E545A, which was in a previously reported hotspot. The other 2 were novel c.1658G>C and c.1659delT frameshift mutations. We conclude that the PI3K/AKT pathway is activated in DLBCL and that PIK3CA is rarely mutated in DLBCL, indicating there could be some other PI3K pathway activation mechanisms operative in DLBCL. PMID- 18382360 TI - Genomic imbalances in urothelial cancer: intratumor heterogeneity versus multifocality. AB - Comparative genomic hybridization and fluorescence in situ hybridization were used to define genetic changes associated with multifocal bladder cancer and to investigate whether the genetic relationship between synchronous urothelial tumors is similar to that observed within different parts of the same tumor. We investigated 8 synchronous urothelial tumors from 3 patients and macroscopically different parts of the same tumor from 2 other patients. The most frequent imbalances were gains of 1q, 2p, and 17q, and losses in 4q. The high number of chromosome imbalances detected in the present report confirms that a high level of chromosome instability could be characteristic of multicentric bladder tumors. Comparative genomic hybridization profiles obtained from independent tumors belonging to the same patient allowed us to elaborate cytogenetic pedigrees portraying the accumulation of chromosome alterations as a form of clonal evolution from a single precursor cell. The analysis of different macroscopic parts of the same tumor allowed us to detect chromosomal heterogeneity and to delineate intratumor clonal evolution. Some chromosome regions that appeared as a gain in one subpopulation were amplified in others indicating a genetic evolution process. Identical processes were observed in different tumors of the same patient. Expansion of chromosome gains and losses between different parts of the same tumor as well as in different tumors of the same patient was also observed. Our results not only provide further evidence of a clonal relationship between different synchronous bladder tumors but also show that the intratumor heterogeneity present in different subpopulations of the same tumor reproduces the behavior of independent synchronous tumors in a same patient. PMID- 18382361 TI - Detection of Mycobacterium tuberculosis by polymerase chain reaction with DNA eluted from aspirate smears of tuberculous lymphadenitis. AB - We have developed and evaluated a polymerase chain reaction (PCR) assay suitable for the detection of Mycobacterium tuberculosis DNA from fine needle aspirate smears of patients with tuberculous lymphadenitis. Air-dried fine needle aspirates of cervical lymph nodes from 98 patients with tuberculous lymphadenitis were studied for cytomorphology, detection of acid fast bacilli by Ziehl-Neelsen staining, culture and nested PCR with IS6110 for mycobacteria on DNA eluted from the dried unstained cytology smear. Twenty aspirate smears with diseases other than tuberculosis were similarly tested as controls. Mycobacterial-DNA was amplified by PCR in 84 (85%) cases and in 1 (5%) control. The mycobacteria could be detected by Ziehl-Neelsen stain and culture in 15 (15.3%) and 24 (24.4%) cases, respectively, whereas both tests were negative in controls. When results were compared with nested PCR on DNA from biopsies from the same case as the gold standard, the sensitivity, specificity, positive and negative predictive values of smear PCR were 85%, 95%, 96%, and 59%, respectively. In conclusion, PCR using dried cytology smear material is feasible and is a simple and sensitive technique for an early and specific diagnosis of M. tuberculosis complex. This is particularly useful when cytology is equivocal and can obviate the use of more invasive procedures. PMID- 18382362 TI - Cell-free DNA fragmentation patterns in amniotic fluid identify genetic abnormalities and changes due to storage. AB - Circulating cell-free DNA (cfDNA) has become a promising biomarker in prenatal diagnosis. However, despite extensive studies in different body fluids, cfDNA predictive value is uncertain owing to the confounding factors that can affect its levels, such as gestational age, maternal weight, smoking status, and medications. Residual fresh and archived amniotic fluid (AF) supernatants were obtained from gravid women (mean gestational age 17 wk) carrying euploid (N=36) and aneuploid (N=29) fetuses, to characterize cfDNA-fragmentation patterns with regard to aneuploidy and storage time (-80 degrees C). AF cfDNA was characterized by the real-time quantitative polymerase chain reaction amplification of glyceraldehyde-3-phosphate dehydrogenase, gel electrophoresis, and pattern recognition of the DNA fragmentation. The distributions of cfDNA fragment lengths were compared using 6 measures that defined the locations and slopes for the first and last peaks, after elimination of the confounding variables. This method allowed for the unique classification of euploid and aneuploid cfDNA samples in AF, which had been matched for storage time. In addition, we showed that archived euploid AF samples gradually lose long cfDNA fragments: this loss accurately distinguishes them from the fresh samples. We present preliminary data using cfDNA-fragmentation patterns, to uniquely distinguish between AF samples of pregnant women with regard to aneuploidy and storage time, independent of gestational age and initial DNA amount. In addition to potential applications in prenatal diagnosis, these data suggest that archived AF samples consist of large amounts of short cfDNA fragments, which are undetectable using standard real-time polymerase chain reaction amplification. PMID- 18382363 TI - High-resolution genotyping of chromosome 8 in colon adenocarcinomas reveals recurrent break point but no gene mutation in the 8p21 region. AB - The prognosis of patients with colorectal cancer is largely determined by the tumor stage. In this respect, colorectal cancer with lymph node metastases has the worst prognosis. Accordingly, there is considerable clinical interest in understanding the genetic mechanisms underlying metastasis formation. The short arm of chromosome 8 is often lost in colorectal cancer and has been associated with the advanced stages. A common region of deletion has been identified in 8p21, and we investigate here the localization of the putative tumor suppressor gene. A series of 683 sporadic microsatellite stability colorectal tumor samples has been genotyped on 12 microsatellite loci encompassing the common deleted region. Allelic losses were identified in 50% of the cases and 10 break points have been evidenced between D8S1734 and D8S1810, reducing the region of interest to D8S1771-D8S131. Among the 21 genes mapped in this interval, 14 candidate genes have been retained for the sequencing analysis of 48 tumors with 8p allelic loss. No mutation was found, suggesting more complex mechanisms of inactivation or side effects of chromosome arm 8q duplication, which might be up-regulating oncogenes not located within the deleted region. PMID- 18382364 TI - Benign metastasizing leiomyoma of the lung: clinicopathologic, immunohistochemical, and micro-RNA analyses. AB - Benign metastasizing leiomyomas are rare tumors, which are typically found in the lungs and, thus, might be confused with leiomyosarcomas. Further, it is not clear whether the term "benign metastasizing leiomyoma" is a misnomer and whether these lesions actually represent low-grade malignant tumors that have a low proliferation index. Micro-RNAs (miRNAs) are small noncoding RNAs, which repress translation. The altered expression of miRNAs has been strongly correlated with the malignant phenotype. In this study, the histologic features, Ki67 index, p53, bcl-2, and miRNA expression were studied in 15 leiomyosarcomas (11 primary lesions and 4 metastases), 8 leiomyomas, and 10 cases of benign metastasizing leiomyoma (9 pulmonary lesions and 1 primary uterine lesion). As expected, the Ki67 index for the benign metastasizing leiomyomas was equivalent to that for the leiomyomas and statistically less than that for the leiomyosarcomas. The mean index was 2.3% (range: 0.9% to 8.8%) for the leiomyomas and 3.4% (range: 0.7% to 8.1%) for the benign metastasizing leiomyomas compared with 28.6% (range: 14.4% to 62.0%) for the leiomyosarcomas (P<0.025). The miRNA, miR-221, which has been associated with a variety of cancers, was detected by in situ hybridization in 13/15 leiomyosarcomas, 0/8 leiomyomas, and 0/10 benign metastasizing leiomyomas. In conclusion, benign metastasizing leiomyomas are indeed most likely benign lesions, and up-regulation of miR-221 expression is an accurate way to differentiate leiomyosarcoma from benign metastasizing leiomyoma. PMID- 18382365 TI - Rapid identification of shiga toxin-producing Escherichia coli (STEC) using electric biochips. AB - Shiga toxin-producing Escherichia coli (STEC) strains are pathogens that cause severe infections, especially in children. Although early data suggested that the STEC phenotype was linked to the O157 serotype, subsequent studies indicated no such strict phenotype-serotype correlation. Thus, it seems that only a test detecting stx1 and stx2 genes (responsible for Shiga toxin production) can provide unambiguous results in STEC identification. Here, we present a system for the rapid detection of the presence of o157, stx1, and stx2 genes, on the basis of an asymmetric polymerase chain reaction and subsequent detection of the amplification products on electric biochips. The whole test lasts less than 100 minutes, and allows for the accurate identification of the stx1 and stx2 genes and the presence of pO157 plasmid in a bacterial colony or in a liquid bacterial culture. The procedure is fully automated, and the only required manual activity is a chip exchange. PMID- 18382366 TI - Cytomegalovirus DNA measurement in blood and plasma using Roche LightCycler CMV quantification reagents. AB - Cytomegalovirus (CMV) is a common cause of morbidity and mortality in immunosuppressed patients. Newly available commercial systems facilitate the measurement of CMV DNA in whole blood or plasma, as a means of detecting and monitoring active disease. We evaluated the performance characteristics of a quantitative polymerase chain reaction that relies on analyte-specific reagents and instruments from Roche Diagnostics. DNA was extracted using a MagNaPure instrument from blood and matched plasma specimens of patients with active CMV disease (defined by another molecular assay) and from controls. Viral load was measured against Roche's standards on a LightCycler instrument, followed by melt curve analysis to confirm product specificity. Dual hybridization probes targeted the CMV UL54 gene and a control sequence that was spiked before extraction. Accuracy and linearity were established using spiked DNA from the Towne-strain CMV. The assay was linear across 6 logs, and it detected CMV DNA in 67/70 blood samples (96%) from patients who were considered to have an active CMV infection, including all 67 viral loads above 208 CMV copies/mL, suggesting that it was sensitive enough to detect clinically significant infections in immunosuppressed patients. Virus levels in plasma correlated reasonably well with the levels in whole blood (r=0.5756), suggesting that levels in either plasma or blood level were indicative of active infection. It was important to verify the calculated values by visualizing the amplification plot and using the melt curve analysis to resolve discrepancies. The LightCycler CMV assay is rapid, sensitive, and linear for quantifying CMV viral load, and it seems to be useful in the diagnosis and monitoring of affected patients. PMID- 18382367 TI - Denaturing high performance liquid chromatography for the detection of microsatellite instability using bethesda and pentaplex marker panels. AB - Microsatellite instability (MSI) is a characteristic molecular phenotype of tumors from the hereditary nonpolyposis colorectal cancer (Lynch) syndrome. Routine MSI screening of tumors in younger patients is an efficient prescreening tool for the population-based detection of Lynch syndrome in the absence of family cancer history. We describe here the optimization of a denaturing high performance liquid chromatography (DHPLC) assay for MSI analysis with the "Bethesda" panel of markers recommended by the National Cancer Institute and with a more recently proposed "pentaplex" panel of 5 mononucleotide repeat markers. By using various polymerase chain reaction primers and tumor DNA samples with known MSI status, each of the 3 standard DHPLC formats tested could correctly identify the MSI status without the "stutter peaks" inherent in the capillary electrophoresis (CE) methods that are currently in use. Dilution experiments showed that the detection limit for MSI using DHPLC was at least 1:100, thus avoiding the need for tumor enrichment by microdissection before analysis. Concordance between CE and DHPLC for the detection of instability in the Bethesda panel markers was 95%. Optimal DHPLC running conditions for the pentaplex mononucleotide panel are also described. In conclusion, DHPLC provides a sensitive and specific alternative for routine MSI analysis that is free of the stutter peaks observed with CE and which can be used with either the Bethesda or pentaplex mononucleotide marker panels. PMID- 18382368 TI - Chromosome 1 analysis in chromophobe renal cell carcinomas with tissue microarray (TMA)-facilitated fluorescence in situ hybridization (FISH) demonstrates loss of 1p/1 which is also present in renal oncocytomas. AB - Morphologic overlap between chromophobe renal cell carcinoma (ChRCC) and renal oncocytomas (RO) has been widely recognized. Whether these tumors are genetically related and represent a spectrum of benign to malignant tumor progression remains an open question. We previously showed by conventional cytogenetics and fluorescent in situ hybridization (FISH) that the most common chromosomal abnormality in RO is loss of chromosome 1 or 1p. In this study, we evaluated chromosome 1 in ChRCC using the same set of FISH probes. Twenty-one ChRCCs from 13 men and 8 women were studied. Formalin-fixed, paraffin-embedded tissue blocks were used to construct tissue microarrays. A subtelomeric 1p36.3 probe was used in tandem with 1q25 probes for FISH studies. The patients ranged in age from 34 to 82 years (mean 62.8 y, median 61 y). FISH analysis showed an abnormal chromosome 1 in 20/21 (95%) ChRCCs as follows: 18 tumors (85%) had loss of entire chromosome 1, 2 tumors (10%) had loss of 1p36.3 only, and 1 tumor (5%) was apparently diploid for chromosome 1. In this study, 95% of ChRCCs showed abnormality of chromosome 1 by FISH. The progression of chromosome 1 abnormalities, from diploid to loss of 1p to loss of entire chromosome, is also present in oncocytomas. These results provide further evidence to support a genetic similarity between chromophobe carcinoma and oncocytoma. Whether abnormalities of chromosome 1 are associated with RO tumorigenesis or its progression to carcinoma requires further studies. PMID- 18382369 TI - Detection of immunoglobulin heavy chain gene rearrangements in classic hodgkin lymphoma using commercially available BIOMED-2 primers. AB - CONTEXT: Classic Hodgkin lymphoma (cHL) is regarded as a clonal B-cell neoplasm. The BIOMED-2 group recently validated a set of immunoglobulin heavy chain (IGH) multiplex primers with high sensitivity in B-cell non-Hodgkin lymphomas. We postulated that after using these primers, a higher proportion of the cHLs would have detectable rearrangements without microdissection. DESIGN: Forty-two patients with cHL were selected. The densities of Reed-Sternberg cells/10 high power field and CD30+ cells/10 high-power field were classified as low, intermediate, or high. The quantities of background CD20+ B cells were classified as low or high. DNA from formalin-fixed, paraffin-embedded sections was used to perform polymerase chain reactions with the InVivoScribe IGH Gene Clonality Assay for ABI detection. Dominant peaks were considered to be monoclonal if they were >3x the height of the polyclonal background, and borderline monoclonal if between 2 and 3x. RESULT: Overall, 10/42 (24%) of the cHL samples were monoclonal, and 7/42 (17%) were borderline monoclonal. Higher densities of CD30+ cells and lower background B cells were statistically correlated with clonality. CONCLUSIONS: The BIOMED-2 primers demonstrate IGH gene clonality in 24% to 40% of cHLs without microdissection. In a subset of the cHL, the IGH gene rearrangement analysis might be useful for diagnosis, but can lead to confusion between cHLs and non Hodgkin lymphomas if used as a discriminative criterion. PMID- 18382370 TI - Germline SDHB mutations are common in patients with apparently sporadic sympathetic paragangliomas. AB - Germline mutations in the genes encoding the B (SDHB) and D (SDHD) subunits of the heterotetrameric protein succinate dehydrogenase (mitochondrial complex II) are important causes of inherited and apparently sporadic paragangliomas. In an effort to further investigate the role of these genes in malignant sympathetic paragangliomas and adrenal pheochromocytomas, we screened a series of tumors for mutations in SDHB and SDHD. Mutation testing was performed on DNA extracted from formalin-fixed, paraffin-embedded tumors and associated normal tissues by polymerase chain reaction amplification and direct sequencing of the coding regions and intron-exon junctions of the SDHB and SDHD genes. Among 16 malignant paragangliomas with proven metastases, 6 (38%) had mutations in SDHB (2 nonsense, 1 splice site, 1 insertion causing a frameshift, and 2 presumably deleterious missense mutations). Probable deleterious SDHB variants were also detected in 5 (45%) of 11 paragangliomas without known metastatic disease (1 splice site, 1 deletion causing a frameshift, and 3 missense changes). In 12 malignant pheochromocytomas, 1 SDHD and no SDHB mutations were identified. The identical SDHB mutation was detected in DNA extracted from accompanying normal tissue for each of the 10 cases on which this analysis was performed. An excess of SDHB mutations in paragangliomas versus pheochromocytomas was found, with no difference in the frequency of mutations in malignant versus benign paragangliomas. The disparate mutational spectra in malignant paragangliomas and pheochromocytomas may reflect differences in underlying tumor biology. PMID- 18382371 TI - Relationship between EBV infection and expression of cellular proteins c-Myc, Bcl 2, and Bax in gastric carcinomas. AB - BACKGROUND: Epstein-Barr virus (EBV) has been related to tumorigenesis in about 10% of all gastric carcinomas. Several studies have demonstrated strong evidence of its involvement in this process, but most of the mechanisms used by the virus to control this process are still unknown. Previous studies in vitro have indicated a relationship between the virus and some cellular genes involved in processes such as proliferation and apoptosis. OBJECTIVE: The aim of the present study was to investigate a possible EBV-induced tumorigenic pathway involving the cellular proteins Bcl-2, Bax, and c-Myc. STUDY DESIGN: One hundred patients of gastric carcinoma, obtained from 2 hospitals in Fortaleza, Brazil were assessed for the presence of EBV by in situ hybridization, for the expression of Bcl-2, Bax, and c-Myc (nuclear and cytoplasmic staining) proteins by immunohistochemistry techniques, and for the apoptotic index. RESULTS: EBV was detected in 8 (8%) patients showing strong staining situated in the nuclei of the tumor cells, 6 of them displaying a diffuse pattern, and 2 demonstrating a focal pattern of staining. The correlation with the immunohistochemistry results demonstrated that none of the EBV-positive cases exhibited Bcl-2 staining. On the other hand, Bax and c-Myc (nuclear) proteins demonstrated a significant positivity index and staining scores (labeling index and H-score) in the EBV positive group; however, the values were lower than those obtained in the EBV negative group, notably for c-Myc nuclear protein. In contrast, the cytoplasmic staining of c-Myc protein revealed slightly higher staining values in the EBV positive group. The balance between Bcl-2 and Bax proteins demonstrated that the majority of the evaluated cases exhibited apoptosis orientation; however, in 62.5% of the EBV-positive cases neither protein was observed. The average apoptotic index was 4.58%, demonstrating a slightly lower average in the EBV positive group. CONCLUSIONS: EBV is not related to the overexpression of Bcl-2 and c-Myc (nuclear) in gastric carcinomas; however, the results point to a possible EBV involvement with the transport mechanisms of the nuclear membrane, resulting in cytoplasmic c-Myc accumulation. The suppression of Bax expression could represent an alternative viral mechanism for inhibition of apoptosis. PMID- 18382372 TI - Real-time quantitative PCR in the diagnosis of tuberculosis in formalin-fixed paraffin-embedded pleural tissue in patients from a high HIV endemic area. AB - The aim of the study was to improve the diagnosis of pleural tuberculosis (TB) based on formalin-fixed biopsies from patients living in high TB and human immunodeficiency virus (HIV) endemic areas. A real-time polymerase chain reaction (real-time PCR) assay targeting a segment of the gene for mycobacterial 65-kd heat shock protein was developed and evaluated on pleural biopsies from 25 patients clinically diagnosed as having TB, on the basis of the good response to treatment, and from 11 controls. A nested polymerase chain reaction (N-PCR) assay for the repetitive genetic sequence insert IS6110, common to Mycobacterium tuberculosis complex organisms, was performed for comparison. When compared with N-PCR, the real-time PCR assay gave a sensitivity and specificity of 83% and 72%, respectively. When compared with clinical diagnosis, the sensitivity and specificity of real-time PCR (68% and 73%, respectively) was comparable with the sensitivity and specificity of the N-PCR assay (64% and 82%, respectively). There were no major differences in the diagnostic validity for the confirmed TB/HIV coinfected patients compared with the results from the whole TB group. In conclusion, the overall accuracy of the real-time PCR assay was comparable with that of the N-PCR and both were equally useful as diagnostic tools in the setting of a HIV coinfection. The real-time PCR has the additional advantage of a short turn-around time, low risk of sample contamination, and offers the possibility to quantify bacterial load, making it a powerful tool for the rapid diagnosis of TB pleuritis. PMID- 18382373 TI - Combined molecular diagnosis of B-cell lymphomas with t(11;14)(q13;q32) or t(14;18)(q32;q21) using multiplex- and long distance inverse-polymerase chain reaction. AB - Translocations t(14;18)(q32;q21) and t(11;14)(q13;q32) are recurrent findings in follicular lymphoma (FL) and mantle cell lymphoma (MCL), respectively. However, the molecular counterparts of these translocations can only be detected in up to 75% of FL and 50% of MCL cases using routine techniques. To improve the efficiency of detection, we first devised a single-tube multiplex-polymerase chain reaction (PCR) assay with primers located within a conserved immunoglobulin heavy chain (IGH) sequence and 5' to the main breakpoint cluster regions of BCL2 and CCND1. Using this assay in 17 FL and 11 MCL diagnostic DNA samples, we readily identified a BCL2-IGH fusion in 65% of FL patients and a CCND1-IGH fusion in 55% of MCL patients. In the remaining cases, we used long distance inverse-PCR to detect BCL2-IGH and CCND1-IGH fusion genes with different BCL2 and CCND1 breakpoint locations. We found additional translocations in 3 patients (17%) with FL and in 4 patients (36%) with MCL. Taken together, we show that multiplex-PCR combined with long distance inverse-PCR detected a t(14;18) in 82% of FL patients and a t(11;14) in 91% of MCL patients, demonstrating that this 2-step protocol is an effective approach for molecular detection of t(11;14) and t(14;18) in B-cell lymphomas. PMID- 18382374 TI - An exploratory look at pediatric cochlear implantation: is earliest always best? AB - OBJECTIVES: Since the advent of cochlear implants, age at implantation has declined as investigators report greater benefit the younger a child is implanted. Infants younger than 12 mos currently are excluded from Food and Drug Administration clinical trials, but have been implanted with Food and Drug Administration-approved devices. With a chance that an infant without profound hearing loss could be implanted because of the limitations of the diagnostic measures used with this population and the potential for additional anesthetic risks to infants younger than 1-yr-old, it is prudent to evaluate benefit in the youngest cochlear implant recipients. The goals of this research were to investigate whether significant gains are made by children implanted before 1-yr old relative to those implanted at later ages, while controlling for potential covariates, and whether there is behavioral evidence for sensitive periods in spoken language development. It was expected that children implanted before age 1 yr would have more advanced spoken language skills than children implanted at later ages; there would be a negative relationship between age at implantation and rate of spoken language development, allowing for an examination of the effects of sensitive periods in spoken language development; and these trends would remain despite accounting for participant characteristics and experiences that might influence spoken language outcomes. DESIGN: Ninety-six children with congenital profound sensorineural hearing loss bilaterally and no additional identified disabilities who were implanted before the age of 4 yrs were stratified into four groups based on age at implantation. Children's spoken language development was followed for at least 2 yrs after device activation. Spoken language scores and rate of development were evaluated along with four covariates (unaided pure-tone average, communication mode, gender, and estimated family income) as a function of age at implantation. RESULTS: In general, the developmental trajectories of children implanted earlier were significantly better than those of children implanted later. However, the advantage of implanting children before 1-yr old versus waiting until the child was between 1 and 2 yrs was small and only was evident in receptive language development, not expressive language or word recognition development. Age at implantation did not significantly influence the rate of the word recognition development, but did influence the rate of both receptive and expressive language acquisition: children implanted earlier in life had faster rates of spoken language acquisition than children implanted later in life. CONCLUSIONS: Although in general earlier cochlear implantation led to better outcomes, there were few differences in outcome between the small sample of six children implanted before 12 mos of age and those implanted at 13 to 24 mos. Significant performance differences remained among the other age groups despite accounting for potential confounds. Further, oral language development progressed faster in children implanted earlier rather than later in of life (up to age 4 yrs), whereas the rate of open-set speech recognition development was similar. Together, the results suggest that there is a sensitive period for spoken language during the first 4 yrs of life, but not necessarily for word recognition development during the same period. PMID- 18382375 TI - Cochlear implantation results in patients with Kearns-Sayre syndrome. AB - OBJECTIVES: To examine the evidence for a cochlear, retrocochlear, or central site of lesion for deafness in two cochlear implant recipients with Kearns-Sayre Syndrome (KSS). DESIGN: Speech perception data and electrically evoked Auditory Brainstem (eABR) and Middle Latency Responses were obtained in two patients with KSS and compared with a group of non-KSS implant recipients. RESULTS: Speech perception data and electrophysiological responses for the patients with KSS were similar to those obtained in non-KSS patients. CONCLUSIONS: Results are consistent with an initial cochlear site of lesion for deafness in KSS, and with relative sparing of the central auditory pathway early in the disease. PMID- 18382377 TI - Transient-evoked otoacoustic emissions in a group of professional singers who have normal pure-tone hearing thresholds. AB - OBJECTIVES: The objective of this study was to determine whether transient-evoked otoacoustic emissions (TEOAEs) measured in a group of normal-hearing professional singers, who were frequently exposed to high-level sound during rehearsals and performances, differed from those measured in age- and gender-matched normal hearing non-singers, who were at minimal risk of hearing loss resulting from excessive sound exposure or other risk factors. DESIGN: Twenty-three normal hearing singers (NH-Ss), 23 normal-hearing controls (NH-Cs), and 9 hearing impaired singers (HI-Ss) were included. Pure-tone audiometry confirmed normal hearing thresholds (>or=15 dB HL) at 0.5, 1.0, 2.0, 3.0, 4.0, 6.0, and 8.0 kHz in NH-Ss and NH-Cs, and confirmed mild, high frequency, sensorineural hearing loss in HI-Ss (HI-Ss were included only to estimate sensitivity and specificity values for preliminary pass or fail criteria that could be used to help identify NH-Ss at risk for music-induced hearing loss). TEOAEs were measured twice in all ears. TEOAE signal to noise ratio (S/N) and reproducibility were examined for the whole wave response, and for frequency bands centered at 1.0, 1.4, 2.0, 2.8, and 4.0 kHz. RESULTS: Moderate to high correlations were found between test and retest TEOAE responses for the three groups. However, absolute test-retest differences revealed standard deviations that were two to three times larger than those reported previously, with the majority of the variability occurring for the 1.0 kHz band. As such, only the best TEOAE response (B-TEOAE) from the two measurements in each ear was used in further analyses, with data from the 1.0 kHz band excluded. With one exception, within-group comparisons of B-TEOAE S/N and reproducibility across ears and gender revealed no statistically significant differences for either NH-Ss or NH-Cs. The only significant within-group difference was between left and right ears of NH-C females for S/Ns measured in the 2.0 kHz band, where median responses from right ears were found to be higher than left ears. Across-group comparisons of B-TEOAEs revealed lower median S/N and reproducibility values for NH-Ss compared with NH-Cs for the whole wave response and 1.4 kHz band. For the 2.0 kHz band, reproducibility was similar for the normal-hearing groups but median S/N was found to be lower for NH-Ss. No significant differences in S/N or reproducibility were found between normal hearing groups for the 2.8 and 4.0 kHz bands. Using data from NH-Cs and HI-Ss to establish sensitivity and specificity values for various TEOAE pass or fail criteria, six preliminary criteria were identified as having sensitivity and specificity values >or=90%. When these criteria were applied to NH-Ss, the number of NH-S ears passing ranged from 57% to 76%, depending on the criteria used to judge the NH-S ears, which translates into 24% to 43% of ears failing. CONCLUSIONS: Although TEOAE responses were measurable in all singers with normal audiometric thresholds, responses were less robust than those of NH-Cs. The findings suggest that subtle cochlear dysfunction can be detected with TEOAE measurement in a subset of normal-hearing professional singers. Although preliminary, the study findings highlight the importance of pass or fail criterion choice on the number of ears that will be identified as "at risk" for music-induced hearing loss. PMID- 18382378 TI - Test-retest repeatability of distortion product otoacoustic emissions. AB - OBJECTIVES: Distortion product otoacoustic emissions (DPOAE) have become part of routine audiological diagnostics. The large scale of clinical DPOAE applications, such as screening of hearing in infants, objective estimation of hearing status, distinction between cochlear and retrocochlear origin of sensorineural hearing loss, exclusion of psychogenic hearing loss, monitoring of hearing during administration of ototoxic drugs, and others illustrates the significance of this audiological tool. In all diagnostic tests, knowledge about the procedure's test retest repeatability is of crucial importance, to allow for distinction between measurement deviations and true physiological or pathological changes in monitoring over time. DESIGN: Measurements of DPOAE were performed in triplicate in 80 normally hearing ears of 40 subjects. Both immediate remeasurements with the ear probe left in place [single-fit mode (SF-mode)] and remeasurements after approximately 5 to 10 days [multiple-fit mode (MF-mode)] were included. DPOAE primary tone levels were varied in 5 dB steps from L2 = 60 to 20 dB SPL (L1 = L2 x 0.4 + 39 dB SPL) and within the frequency range f2 = 1 to 6 kHz. Repeatability of DPOAE was evaluated by the standard error of measurement (Sm), reliability (Cronbach alpha), absolute differences between measurements, 95% confidence intervals, and repeatability standard deviations. RESULTS: Sm averaged 0.67 dB over all frequencies and primary tone levels in the SF-mode, and 1.44 dB in the MF-mode, respectively. As expected, test-retest repeatability declined with decreasing primary tone levels; however, repeatability values were still mostly satisfactory with the lower primary tone levels. For the exemplary primary tone level combination of L1/L2 = 63/60 dB SPL, which is close to common clinical paradigms, the difference between two DPOAE measurements under the reported test conditions could be considered statistically significant (p = 0.05) if it exceeded 0.7 to 1.3 dB in the range 1 to 5 kHz and 2.3 dB for 6 kHz in the SF mode, when compared with 1.8 to 2.7 dB for 1 to 5 kHz and 3.7 dB for 6 kHz in the MF-mode. Signal to noise ratio (SNR) did not seem to have a large influence on repeatability, as long as SNR was within 6 to 35 dB, which covers the range of most clinical DPOAE measurements. CONCLUSIONS: The DPOAE-test-retest study presented here is to our knowledge the first, which combines variation of primary tone levels, assessment of both SF- and MF-modes, and comparison of the two modalities within the same subjects. Although the measurements were conducted under practical conditions resembling the clinical setting, repeatability was generally good. The widely used minimum SNR of 6 dB seems to be a recommendable criterion when considering both practicability and measurement quality under clinical conditions. The current findings underline the suitability of DPOAE as a monitoring tool of cochlear status over time. The data are intended to assist the clinician and the scientist in the correct interpretation of DPOAE level changes in the test-retest situation. PMID- 18382381 TI - Neonatal necrotizing fasciitis due to community-acquired methicillin resistant Staphylococcus aureus. PMID- 18382382 TI - Cytokine gene polymorphism in respiratory syncytial virus bronchiolitis. PMID- 18382383 TI - Congenital asplenia and severe visceral toxocariasis. PMID- 18382384 TI - An adenovirus type F41 outbreak in a pediatric bone marrow transplant unit: analysis of clinical impact and preventive strategies. AB - BACKGROUND: Adenovirus (HAdV) was identified in blood and stool specimens from 6 children on a pediatric bone marrow transplant (BMT) unit within 2 weeks. Two further adenovirus positive patients were identified in other areas of the childrens' hospital. The study aimed to determine the clinical course of different HAdV subtypes and to investigate whether the cluster was caused by nosocomial transmission or by endogenous reactivation. METHODS: Descriptive epidemiologic investigation was performed reviewing patients' charts. Molecular typing of identified adenovirus-DNA was performed by partial sequencing of the hexon gene. RESULTS: In 6 of 8 patients, HAdV-F41 was detected in feces. All but 1 patient presented with vomiting or diarrhea and all were treated with cidofovir. In 4 patients transmissions of HAdV-F41 within the hematological department were probable whereas 2 children on the BMT ward reactivated HAdV-C1 and -C2, respectively. HAdV-F41 was shed in feces for up to 64 days after onset of clinical symptoms. HAdV-F41 DNA in blood reached a maximum of 2 x 10(5) copies/mL. One patient harbored two HAdV types simultaneously, HAdV-F41 in feces and HAdV-C2 in blood samples. HAdV-C2 reached high virus concentrations in blood (4 x 10(9) copies/mL) and led to the only fatal case. Although the HAdV-F41 outbreak involving 6 children led to gastroenteritis and may also have been associated with mild hepatitis, coincidental, endogeneous reactivations of other HAdV types (C1 and C2) led to a more severe course. CONCLUSIONS: HAdV typing is essential both for the prognosis and for distinguishing between transmission or endogenous reactivation. Applying HAdV-specific infection control measures is crucial to prevent transmission. PMID- 18382387 TI - Antibody response to hepatitis A immunization among human immunodeficiency virus infected children and adolescents. AB - Seventy-one of 84 human immunodeficiency virus (HIV)-infected children [84.5% (95% confidence interval: 75-91.5%)] were hepatitis A virus (HAV) seropositive after 2 doses of HAV vaccine. Higher CD4% and HIV suppression were significantly associated with increased HAV seropositivity rate. In multivariate analysis, CD4 >or=25% and young age were independent predictors of HAV seropositivity. Of 7 children given a third HAV vaccine dose because of negative HAV antibody after 2 doses, 2 (29%) became seropositive. PMID- 18382386 TI - Long-term safety and effectiveness of ritonavir, nelfinavir, and lopinavir/ritonavir in antiretroviral-experienced HIV-infected children. AB - AIM: To evaluate the long-term safety and effectiveness of ritonavir, nelfinavir, and lopinavir/ritonavir in antiretroviral-experienced, initially protease inhibitor (PI)-naive, human immunodeficiency virus (HIV)-1-infected children. METHODS: HIV-1-infected children enrolled in the Swiss Mother and Child HIV Cohort Study were eligible for this observational cohort study if they received at least 1 PI of interest between March 1996 and October 2003: ritonavir, nelfinavir, or lopinavir/ritonavir. Data regarding demographics, clinical disease and antiretroviral treatment history, HIV-1 RNA copies/mL, CD4 T-cell counts [absolute (cells/microL) and percentages (%)], adverse events, clinical laboratory values, reasons for discontinuation of PIs, and concomitant medications were extracted from the database for PI-naive (first-line) and PI experienced (second- or higher-line) PI use. RESULTS: The total duration of ritonavir, nelfinavir, and lopinavir/ritonavir use for 133 HIV-1-infected children was 163.8, 235.0, and 46.1 patient-years, respectively. In an on treatment analysis, first-line therapy with any of the PIs significantly reduced HIV-1 concentrations and increased CD4 T-cell counts and percentages from baseline throughout the 288-week study (P >. The angiographic examination of the vessels of the lower extremities was carriedout on the angiographic complex manufactured by the Company Siemens Multistar T.O.P. The volume of the filling of the vessels of the lower limbs was evaluated by the parameters of the rheogram registered on the rheograph 4RG-1A. It was determined that the inclusion of mexicor (5-6% ml intravenously dropwise, 300 mg/day) into the standard therapeutic regimen significantly increased the efficacy of the conventional management of atherosclerosis obliterans of the arteries of the lower limbs, accelerating regression of the main clinical symptoms of ischaemia, increasing the distance of pain-free walking, correcting dyslipidaemia, and excreting an antioxidant action. PMID- 18382391 TI - [Rhythmic pneumocompression for treatment of patients with chronic obliterating diseases of the lower-limb arteries]. AB - A total of 68 patients suffering from chronic obliterating diseases of the lower limb arteries (CODLLA) under-went treatment with rhythmic pneumocompression (RPC) in the descending <> mode with a simultaneous relief of pressure in the sections of the cuffs. The values of the pneumatic action exceeded the APS determined on the shoulder by 10-20 mm Hg. The therapeutic course consisted of 10 15 sessions. Clinical improvement commenced to be observed as soon as after the first session of RPC: the patients were found to have decreased pain, and increased distance of pain-free walking. Based upon the parameters of the dynamics of the cutaneous temperature on the feet, transcutaneous determination of pO2, dynamic pulse oximetry, acid-base balance, and the indices of the blood gaseous composition, the authors have hereby arrived at the conclusion that the therapeutic impact of the RPC in the mode of the descending <> with the values of the pneumatic impact exceeding the APS was related to the opening of the capillary bed and stimulation of the metabolic processes. The method may therefore be safely recommended to be used in patients presenting with CODLLA, especially in the presence of distal lesions to the arteries of the lower extremities, as a preoperative preparation of the vascular bed. PMID- 18382392 TI - [Administration of prostaglandins Ei and immunotherapy in management of various genesis trophic ulcers]. AB - OBJECTIVE: To study the dynamics of the indices of microcirculation and the immune status in patients suffering from indolent trophic ulcers with the preserved arterial blood flow, as well as a possibility of treating them by means of PGEi-group preparations combined with immunocorrection. PATIENTS AND METHODS: From 2002 to 2006 in the Clinic of Vascular Surgery of Municipal Clinical Hospital No 3 of the city of Chelyabinsk, we examined a total of 158 patients suffering from indolent trophic ulcers of the lower extremities with the preserved arterial blood flow (venous ulcers and cutaneous angiitis). Also studied were the immune status and tissue perfusion indices, cytology of the ulcers' surface. Treatment was carried out using <> and immunocorrecting therapy (prednisolone and delagil). RESULTS: At baseline, all the patients were found to have decreased indices of tissue perfusion, with a direct correlation observed between lowered tissue perfusion and the duration of lingering trophic disorders. Venous pathology and dermohypodermal angiitis were observed to be accompanied and followed by similar alterations in the immune status. Upon completion of the therapeutic course, we noted a statistically significant increase in tissue perfusion in the groups studied. Clinically noted was decreased oedema of the lower extremities, disappearance of the pain syndrome, and positive dynamics of the healing of trophic ulcerative defects. Complete epithelisation was achieved in 100% of the patients with venous insufficiency and the initial surface of the ulcer up to 60 cm2. In patients presenting with vasculitis and the baseline area of the trophic ulcer up to 38 cm2 four months after therapy ulcers were noted to epithelialize completely in 61% of cases. CONCLUSION: Combined therapy of various-genesis trophic ulcers with the preserved major blood flow by prostaglandins Ei preparations (vasaprostan) and immunocorrecting preparations (prednisolone and delagil) turned out pathogenetically substantiated and clinically efficient. The authors have for the first time proposed herein using the drugs of PGEi -group in therapy of venous trophic ulcers. PMID- 18382393 TI - Endovascular treatment of a renal artery stenosis: techniques, indications, results. Role of embolic protection devices (part 3). PMID- 18382394 TI - [Paclitaxel-eluting stents in treatment of stenoses and chronic occlusions of coronary arteries]. AB - The present review summarises the data obtained by analysing the publications dedicated to using paclitaxel-eluting stents in management of coronary heart disease. The stents containing paclitaxel (taxol) proved to have high clinical efficacy in both single and multiple stenoses of the coronary arteries. The feasibility of using taxol-eluting stents in patients suffering from diabetes mellitus and in patients diagnosed with chronic coronary occlusion does not admit of any doubt. However, the rate of restenoses development following implantation of the paclitaxel-eluting stents is higher than that after using the syrolimus releasing stents. Unsolved as yet remains the issue concerning the efficacy of the paclitaxel-eluting stents intended for treatment of restenoses inside the stent, as well as concerning the feasibility of using these stents for stenting the main trunk of the left coronary artery. PMID- 18382395 TI - [Assessment of therapeutic efficacy in thromboses of the inferior vena cava by means of colour duplex scanning]. AB - A total of two hundred and twenty-eight 18-to-83-year-old patients diagnosed with thrombotic occlusion of the lower-limb deep veins were examined in order to evaluate the possibilities of colour duplex angioscanning as a method aimed at monitoring both the course of and efficacy of treatment for acute venous thrombosis, as well as at preventing thromboembolic complications. Studied were the localization, incidence rate, and complications of the course of various types of venous thrombosis. It was demonstrated that an uncomplicated course the disease was observed in more than 80% of cases. At the same time, using dynamic ultrasonic monitoring made it possible to promptly detect progression of the pathological process in 3.5% of cases, and to introduce due amendments in the treatment policy being carried out. Also shown were the possibilities of the colour duplex scanning (CDS) technique in assessing the outcomes of the endovascular prevention of pulmonary thromboembolism (PTE) by means of implantation of the cava filter. The CDS turned out efficient in diagnosing the development of such complications as dislocation of the cava-filter and thrombus formation in the area of the cava filter. It was determined that the terms of fixation of the embolus-hazardous (floating) thrombus varied from 6 days to 6 months, which is of great importance in selecting the optimal period for temporary endovascular prevention of PTE. PMID- 18382396 TI - [Duplex scanning in study of haemodynamics of genital organs during pregnancy]. AB - The article deals with the findings of studying the haemodynamics of the genital organs by means of duplex scanning in a total of 32 pregnant women presenting with varicosity of genital veins. It was revealed that progression of gestation leads to increasing severity of symptoms and enlarged number of the signs of varicose disease (VD) of the genital organs, with the peak manifestation thereof being observed by the term of delivery. In the postpartum period, noted was a significant decrease in the degree and incidence rate of the major symptoms of VD by day 5-7. PMID- 18382397 TI - [Unadequate ostial ligation of the great saphenous vein as the cause of varicose disease reccurence]. AB - Authors investigated the frequency of varicose disease recurrence after crossectomy, classified clinical types and proposed the tactics of treatment of this group of patients. 258 patients with varicose disease recurrence were examined and operated. In 179 cases (69,4%) there were found differ pathological changes in the place of saphenofemoral junction, which were collected in several groups. Pathological stump of the great saphenous vein was in 155 lower limbs (60,1%). 12 patients (4,7%) had not ligated saphenofemoral junction. And in 12 patients (4,7%) there were found signs of neoangiogenesis. We proposed the scheme of tactical actions in cases of pathological changes in the place of saphenofemoral junction. The absolute indication for surgery is present saphenofemoral junction and trunk of the great saphenous vein. Furthermore, recection of vein's stump is necessary if its length is more 3 cm, that always combined with varicose tranformation of the varicosities. If the linear size of stump is from 1 to 3 cm its resection have to be done only in case of considerable varicosities. In not severe lesions of varicosities it is enough to perform sclerotherapy or mini-phlebectomy of involved vessels. There is not necessary to surgery if the stump is short (less 1 cm). In this case the phlebosclerotherapy or mini-phlebectomy of varicosities are expedient. The method of choice for removal of neoangiogenesis is phlebosclerotherapy. Our results showed that treatment of patients with varicose disease recurrence is in special phlebological centers or vascular departments and improvement of technique of Troyanov-Trendelenburg operation. PMID- 18382398 TI - [A differentiated approach to management of small-pelvis varicosis in women]. AB - In our clinic, we examined and treated a total of 32 women presenting with small pelvis varicosis (SPV) and suffering from chronic pelvic pain appearing after prolonged static loads and physical activity, dysuric impairments, dyspareunia, and painful hypermenorrhoea. A Doppler ultrasonographic study and phlebography revealed SPV in them too. Appropriate treatment was carried out in a differentiated manner depending upon the stage of the disease and degree of clinical manifestations. Eighteen women with stage 1-2a SPV underwent comprehensive conservative treatment including venotropic preparations, microcirculation-improving agents, drugs influencing systemic enzymopathy, and physiotherapy. The remaining 14 women with stage 2b-3 SPV were subjected to roentgenovascular occlusion of the ovarian veins by means of sclerosing agents and Guianturco-type metal spirals. At 2-3 months after conservative and surgical treatment. 23 patients (72%) reported disappearance of pelvic pain; five women (16 %) were found to have their pain subdued; in four subjects (12.5%) pain did not disappear. During a 4-year period after treatment, 27 women (84%) had reportedly no dysuric events, five women (16%) at 2-2.5 years after treatment appeared to have developed exacerbations of chronic cystitis, two patients (6%) turned out to have periodically (2-3 times a year) been experiencing frequent painful urination. Besides, we carried out a morphological examination of the urinary-bladder walls on the post-mortem materials from 15 (unrelated) cases of accidental death, having revealed varisosis of the small pelvis. The findings obtained therein also strongly suggested that lingering impairment of the venous outflow from the small pelvis had led to development of a chronic inflammatory process in the urinary-bladder wall. PMID- 18382399 TI - [Reconstructive operations in unilateral occlusions of the iliac vein]. AB - Analysed herein are the findings of the examination and outcomes of surgical management of 48 patients with unilateral occlusion of the iliac veins. Of these, grade I chronic venous insufficiency (CVI) was observed in 12 patients and grade II SVI in 28 subjects. Grade III CVI's with trophic ulcers were diagnosed in 8 patients. In a further eight patients, the primary cause of thrombosis had been catheterization of the femoral vein in childhood. Nine patients developed thrombophlebitis following the endured enteric fever, with four patients having developed it after an injury, four - after an operation on the abdominal cavity organs, and four - after delivery. In the rest 19 patients, we failed to reveal the cause of thrombophlebitis. The venous haemodynamics was studied by means of the following methods: Doppler ultrasonography, duplex scanning and phlebography. The latter was used by the indications, with the duplex scanning technique having proved to play the leading role. The patients with postthrombotic lesions of several segments appeared to have a severer degree of CVI. The venous circulation was more compensated in the well-developed suprapubic collateral network. The patients were subjected to two types of surgical interventions: the Palm-Esperon operation with and without the establishment of the AV fistula (26), and correction of the suprapubic collaterals (21). The indication for establishing the AV fistula was the difference of pressure in the femoral veins of not less than 50 mm Hg. The correction of the suprapubic collaterals was carried out in considerably varicose-dilated saphenous veins of the suprapubic area. Disseminated lesions of the deep veins and secondary alterations in the tibial veins were additionally managed and coped with by the appropriate correcting operations performed on the crus. Postoperative complications were observed in two (4.2%) patients: thrombosis of the femoral-iliac shunt and of the corrected suprapubic vein. Favourable outcomes were obtained in 46 (95.8%) patients. PMID- 18382400 TI - [Venectomy under local tumescent anaesthesia]. AB - The authors have analysed the findings of the examination and outcomes of surgical management of 142 patients with lower limb varicosity (a total of 160 lower extremities). Of theses, there were 52 (36.6%) men, and 90 (63.4%) women. The patients' age varied from 18 to 74 years old (mean age 44+/-11.1 years). The average duration of the disease amounted to 8.3+/-5.2 years. The distribution according to the CEAP clinical classes was as follows: C2 - 90 patients (63.4 %), C3 - 43 (30.3 %), C4 - 6 (4.2 %), C5 - 3 (2.1 %). The proportion of the patients with large varicose veins (more than 1 cm in diameter) amounted to 31.25%. Lesions of the bed of the great saphenous vein (GSV) were observed on 139 lower limbs (86.8%), of the small saphenous vein (SSV) - on 19 lower limbs (11.9%), and of both the GSV and SSV - on two lower extremities (1.3%). In all the cases, the operation was performed under local tumescent anaesthesia in outpatient conditions. The anaesthetic mixture was based on 0.2 - 0.5% ropivacain. The subjective score of the anaesthesia quality averaged 9.6 points. Immediate postoperative complications were observed in 1.4% of the patients (lymphorrhoea and wound process). Side effects of anaesthesia in the form of a vegetovascular reaction, transient cutaneous paresthesia, and a skin reaction were noted in 8.4% of the patients. Local postoperative haematomas were observed in 9.1% of cases. A total of 83.8% of the patients did not use tabletted analgetics in the postoperative period. The subjective assessment of the surgical intervention performed and postoperative outcomes achieved was excellent to good in 98.6% of the patients. PMID- 18382401 TI - [Assessing efficacy of combined use of endovasal laser congulation and echosclerotherapy in comprehensive management of complicated forms of varicose disease]. AB - The article seals with the findings obtained in management of 96 patients diagnosed with varicose disease in the stage of trophic disordwrs treated by means of rndovasal laser obliteration combined with echosclerotherapy. Presented herein are peculiarities of carrying out laser coagulation of the greater and minor saphenous veins, as well as incomperetent perforating veins without ligation of the sapheno-femoral and sapheno-popliteal junction. The duration of the follow up of the patients amounted oj the patients amounted to three years. In all the cases we managed to achive persistent epitheliali-sation of the ulcerative defect and attain considerable improve-ment of the patients' quality of life by virtue of shortening the terms of medical and social rehabilitation and heightening the cosmetic effect. Endovasal laser coagulation combined with sclertherapy is a method of choice in a comprensive treatment of complication forms of varicose disease. PMID- 18382402 TI - [State of the greater saphenous vein following intraoperative truncal scleroobliteration in varicose disease]. AB - The authors have analyzed the outcomes of intraoperative truncal scleroobliteration of the greater saphenous vein, which was used instead of stripping in patients presenting with varicose disease of the lower limbs. Obtained were three variants of the condition of the vessel concerned following the intervention: complete obliteration, preserved lumen, and alternating portions of obliteration and patency. It was determined that varicosity of saphenous veins and incompetent perforating vein of Dodd were more often encountered in preserved patency of the trunk of the greater saphenous vein on the femur, which required additional treatment. Tested were methods of non surgical correction of the surgical outcomes by means of echosclerotherapy. PMID- 18382403 TI - [Results of administering Phlebodia 600 in fertile women with orthostasis dependent phlebopathy]. AB - Presented herein are the findings of a prospective, non-comparative study of the clinical efficacy of semisynthetic diosmine (preparation Phlebodia 600) in patients presenting withCEAP class 0-1 chronic venous insufficiency (CVI) manifesting itself as orthostasis-dependent phlebopathy. Based on a total of 67 case reports, it was shown that Phlebodia 600 exerted a positive effect on the functional symptoms of CVI, oedematic and convulsive syndromes. A conclusion was made of a high efficacy and safety of the drug in treatment of initial-grade CVI in fertile women with orthostasis-dependent phlebopathy, including hormone induced variants. Noted was positive dynamics of the preparation with the necessity of continuing to take oral contraceptives by the medical indications. PMID- 18382404 TI - [Reconstructive surgery of carotid arteries in patients over 70 years old]. AB - Analysed herein are the immediate outcomes of primary reconstructive operations on carotid arteries performed in conditions of regional anaesthesia in patients over 70 years old and those younger than 60 years old. In each group, we singled out the risk factors of operative surgical treatment. The group over 70 years included a total of 55 patients who endured a total of 61 carotid operations. In the group of patients younger than 60 years, we performed a total of 237 operations on the carotid arteries in 223 patients. The perioperative mortality rate in the over-70-year-old group patients amounted to 1.64% (1 death), and in the group of the patients younger than 60 years old it equalled 1.27% (3 deaths), P>0.5. In the group of the patients over 70 years old, death was caused by ischaemic stroke in the carotid bed operated on. The causes of death in the group of the patients younger than 60 years old included ipsilateral stroke in two patients and acute myocardial infarction in one patient. Non-lethal homolateral stroke was observed in one (1.64%) patient over 70 years and in four (1.69%) patients younger than 60 years old (P>0.5). The <> index amounted to 3.28% and 2.96% in the groups of the patients above 70 and those under 60 years of age, respectively. The peculiarities, observed in the patients over 70 years old were as follows: a high incidence rate of concomitant diseases of the vital systems, a bilateral and an embogenic pattern of the carotid pathology. In the group of patients under 60 years predominating were those with endured stroke - 97 (43.49%) patients; of these, 64 (65.98%) had embogenic atherosclerotic plaques. The immediate results of the reconstructive operations performed on the carotid arteries in patients over 70 and those under 60 years have turned out comparable and quite acceptable. PMID- 18382405 TI - [A comparative analysis of long-term outcomes of femoropoplietal autovenous bypass <> grafting and the reversed vein]. AB - Presented herein is a comparative analysis of the remote results obtained ofter a total of 215 operations of femoropoplietal autovenous bypass grafting performed by a reversed vein (n=105) and <> technique (n=110). The patients from the both groups matched well by all criteria of the comparative analysis. Over a 5-year period, the cumulative patency rate of the <> grafts amounted to 56.4%, and that by the reversed autovein equalled 47.6 %. The preservation rate in the both groups during the same time period amounted to 77.3 % and 67.7 %, respectively. The main factor exerting a statistically reliable influence on the patency of the femoropoplietal grafts was found to be the level of the establishment of the distal anastomosis. When the distal anastomosis was established, above the knee-joint fissure, no statistically significant differences in the patency of the grafts performed by of the reversed autovein and those carried out according to the <> technique were revealed. Placing the distal anastomosis below the knee-joint fissure appeared to offer apparent advantages of autovenous grafting performed according to the <> technique. PMID- 18382406 TI - [Remote results of ultrasonic endarterectomy]. AB - The authors carried out a comparative retrospective study of the efficacy of ultrasonic endarterectomy used for the restoration of the blood flow in patients diagnosed with atherosclerotic lesions of the femoral artery. The long-term outcomes of the operation (with the duration of the follow-up period amounting to 16 years) were analysed in a total of 535 patients. Based on the variety of different types of the operations performed, the patients were subdivided into three groups: Group I consisted of 208 subjects having undergone ultrasound mediated endarterectomy, Group II comprised 244 patients having endured femoropopliteal bypass grafting with the reversed autovein above the knee joint; and Group III was composed of 83 patients following mechanical endarterectomy. The groups of the patients analysed were matched by sex, age, duration of the disease, the prevalence rate of concomitant diseases, and the degree of chronic ischaemia of the extremity involved. The criteria for assessment of the remote outcomes were the indices of the patency of the reconstructed vessels and limb salvage. It was determined that ultrasonic endarterectomy was accompanied and followed by better results of the patency as compared with mechanical endarterectomy, being however inferior to the operation of the femoropophteai bypass grafting by the criteria applied. No differences in the parameters of limb salvage after various modalities of the restoration of the blood flow were observed. PMID- 18382407 TI - Surgical treatment of a female patient with Turner's syndrome and a giant dissecting aneurysm of the ascending artery which occurred after two operations for aortic coarctation. AB - This paper reports a female patient with. Turner's syndrome and a giant aneurysm of the ascending aorta with pronounced aortic insufficiency in the presence of the bicuspid aortic valve, type II aortic dissection that occurred after two operations: resection of aortic coarctation with end-to-end anstomosis, recoarctation syndrome and aortic ascenoing-descending bypass grafting via the right-sided thoracotomy. The patient underwent prosthetics of the ascending aorta and aortic valve with ascending-descending repeated bypass grafting under conditions of complete peripheral cardiopulmonary bypass, hypothermia (22 degrees C) antegrade brain perfusion via the right subclavian artery. PMID- 18382408 TI - [Successful surgical treatment of veritable internal carotid aneurysm due king king]. AB - In the report authors had shown the clinical observation (case) of surgical treatment of the patient with veritable lateral aneurysm of right internal carotid artery appeared due of its king-king. PMID- 18382409 TI - [Femoropopliteal bypass above the knee-joint fissure by means of a vascular prosthesis made of PTFE: choosing an appropriate diameter of the prosthesis]. AB - Presented herein are the outcomes of a total of 80 operations of femoropopliteal grafting above the knee-joint fissure, carried out using a vascular prosthesis made of polytetrafluoroethylene (PTFE) in patients diagnosed with atherosclerotic lesions of the arteries of the lower extremities over the period between February 1995 and December 2005 at the Department of Vascular Surgery of the A. V. Vishnevsky Institute of Surgery of the Russian Academy of Medical Sciences. Fifty percent of the patients were operated on for grade II B ischemia, while 13.7% of the patients suffered from grade III ischaemia of the lower limb, and 29 (36.3%) subjects were found to have grade IV ischaemia. The condition of the distal bed was considered <> in 41.2% of the patients, being defined as <> in 48.5%, and as "poor" in 10.3%. The operative intervention was carried out according to the conventionally accepted technique. 50% of the reconstructions were performed using the prosthesis 8 mm in diameter, and 50% - using that measuring 6 mm in diameter. A statistically significant impact of the diameter of the prosthesis used on the patency of the graft in the remote postoperative period was revealed: with the prosthesis 8-mm in diameter, the patency after 12, 30 and 60 months amounted to 93%, 64% and 51%, respectively, while with the transplant measuring 6 mm in diameter, these indices at the above mentioned time intervals did not exceed 62%, 44% and 31%, respectively (P=0.004). PMID- 18382410 TI - Effects of toluene-2,4-diamine on red sea bream, Pagrus major: biochemical and histological evaluation. AB - The subacute toxicity of toluene-2,4-diamine (2,4-TDA) on marine fish was investigated in laboratory toxicity tests using red sea bream, Pagrus major. The fish were exposed to subacute concentrations of 2,4-TDA (measured concentrations: 0.0628, 0.234 and 0.878 mg/L) during the 14-day toxicity test and the effects on growth as well as on some biochemical parameters in the plasma, gills, liver and kidney structures were studied. The body weight and body length of the red sea bream exposed to the highest concentration were lower than those of the control. This study also showed a tendency towards decreases in three hematological parameters, i.e., the number of red blood cells, hemoglobin level and hematocrit, as well as blood chemical parameters in the plasma, i.e., the levels of total cholesterol, triglyceride, total protein and albumin, of fish exposed to the high concentrations. These findings were attributable to the reduced feeding of fish exposed to this concentration. In addition, cytoplasmic changes were observed in the cells of fish exposed to the highest concentration. It is suggested that the changes were caused by the decrease in feed efficiency derived from the exposure to 2,4-TDA. PMID- 18382411 TI - Mass balance study of nonylphenol ethoxylates and their metabolites in an urban river contaminated by nonylphenol. AB - In previous studies, an urban river called Kamo River was found to be polluted with nonylphenol (NP) by a rubber product manufacturing factory. To determine the contribution of the factory effluent to the river pollution, a mass balance study was conducted for NP, nonylphenol ethoxylates (NPnEOs, n = 1-15), and nonylphenol carboxylates (NPmECs, m = 1-10) in the waters of the river and its 10 inflow channels. The total concentration of nonylphenolic compounds (T-NPCs) in the river water was 4.6 nM/L at the upstream sampling point and 54.6 nM/L at the downstream sampling point. T-NPCs concentration ranged from 0.4 to 487.1 nM/L in the waters of the inflow channels connected to the targeted river section, and the highest value was observed in the water of the inflow channel which receives the factory effluent. The mass flows of T-NPCs were 458.0 mM/day for Input (the upstream sampling point and outlets of 10 inflow channels) and 828.2 mM/day for Output (the downstream sampling point). The mass flow of the highly polluted inflow channel comprised 82.4% of Input. In comparison with the mass balance of chloride ion, the mass flow of T-NPCs in Output was still higher than that in Input. This phenomenon was attributed to the high mass flows of NP and NPnEOs (n = 1-3) in Output, and their potential source was determined to be the river sediment. PMID- 18382412 TI - Pesticide usage and its association with health symptoms among farmers in rural villages in West Java, Indonesia. AB - The improper handling of pesticides in agriculture has caused serious health problems in many developing countries. In this study, we report the pesticide usage condition among Indonesian farmers and its association with symptoms of pesticide toxicity. A questionnaire survey on personal history regarding agricultural labor, pesticide storage and disposal, pesticide use and health history was conducted using a structured questionnaire in rural Sundanese villages in West Java, Indonesia. The most frequently used pesticides included dithiocarbamates, pyrethroids and organophosphates. In approximately 80% of sprayings, category II pesticides (World Health Organization (WHO) categorization; "moderately hazardous") were used. Many of the subject farmers worked in a highly unsafe occupational environment; protective measures and safe handling were rarely observed, whereas smoking and drinking during spraying were frequently practiced. Correlation analysis revealed that farmers who wore a long sleeve shirt and headgear showed health symptoms less frequently. Moreover, farmers who had skin contact with the spray solution during measuring or mixing (excluding the hands), who wore wet clothing (skin exposure to pesticide), and who smoked and rubbed their eyes during spraying showed more symptoms. Among these factors, headgear use, wearing wet clothing (skin exposure to pesticide), and smoking during spraying were the significant determining factors for developing health symptoms. Preventing such behaviors will be an effective method of reducing health problems among the subject farmers. PMID- 18382413 TI - Toxicity assessment of wastewater by proteomics analysis. AB - The effects of effluent from a wastewater treatment plant (EWWTP) on intestinal epithelial Caco-2 cells, a human intestinal epithelial cell line derived from a human colon carcinoma, were investigated. Previous studies have shown that the wastewater constituents nonylphenol and lipopolysaccharide (LPS) induce the overexpression of specific proteins (galectin-3, glutathione S-transferase A2 subunit, peroxiredoxin-1, and heat shock protein 90, beta (HSP90b)). In this study, the first screening of EWWTP was carried out using the HSP47-transformed cell assay, which is a highly sensitive toxicity assay. From the results of proteomics analysis of human intestinal Caco-2 cells treated with EWWTP, we found the overexpression of specific proteins, namely, elongation factor 1beta and enolase 1. These results suggest that specific proteins can be used as biomarkers for the risk assessment of water and wastewater. PMID- 18382414 TI - Assessment of estrogenic activity in Tunisian water and wastewater by E-screen assay. AB - Wastewater and surface water samples from three wastewater treatment plants (WWTPs) and three rivers in Tunisia were assayed for estrogenic activity using the E-screen assay and enzyme-linked immunosorbent assay (ELISA). Results showed that all the Tunisian raw wastewater samples as well as the Roriche river water sample induced a strong proliferative response in human MCF-7 breast cancer cells. Tunisian raw wastewater had an average 17beta-estradiol content of 2,705.4 pg/ml, whereas that of the Roriche river was 36.7 pg/ml, which is sufficient for inducing endocrine-mediated responses in aquatic organisms. Results further showed that the Mornag WWTP, which uses the activated-sludge treatment system, has a higher estrogen removal efficiency than the stabilization ponds of the Gammart and pilot WWTPs. This study, which is the first of such studies in Tunisia, and probably the first in the North African region, underscores the need to detect and monitor the estrogenic activity of water and wastewater, given the scarcity of water in Tunisia and the detrimental impact of endocrine-disrupting compounds on the physiology of both animals and humans. PMID- 18382415 TI - Effects of light condition after simulated acid snow stress on leaves of winter wheat. AB - Winter plants regrow after freeze-thawing in acidic meltwater from the acid-snow layer in early winter or early spring. In this study, the responses of cold acclimated wheat seedlings to different light conditions during the regrowth period after simulated acid snow (SAS) stress were investigated. After freeze thawing in sulfuric acid (SAS stress) of pH 2.0, dry weight and the maximal quantum yield of photosystem II (PSII) decreased more in mature leaves than in young leaves. In a subsequent regrowth period under light condition, dry weight, relative water content, and the maximal quantum yield of PSII were severely affected in mature leaves but were only slightly affected in SAS (pH 2.0) stressed young leaves. The levels of membrane lipid peroxidation and hydrogen peroxide in mature leaves of SAS (pH 2.0)-stressed seedlings were significantly higher than those in young leaves during the regrowth period under light condition. The superoxide dismutase activity in young leaves was higher than that in mature leaves during the regrowth period. These results indicate that mature leaves of seedlings during the snow melt season are more sensitive than young leaves to photooxidative stress because of their low acid snow stress tolerance and low capacity for the detoxification of superoxide. PMID- 18382416 TI - Preliminary ecological risk assessment of butylparaben and benzylparaben -1. Removal efficiency in wastewater treatment, acute/chronic toxicity for aquatic organisms, and effects on medaka gene expression. AB - Butylparaben and benzylparaben, used as preservatives mainly in cosmetic products, have recently been found to be weakly estrogenic. Batch activated sludge treatment and batch chlorination were carried out to roughly determine the removal efficiency of a wastewater treatment plant. Combining the removal efficiency with the estimated annual consumption and the unaltered excretion ratio, the maximum predicted environmental concentration (PEC) was estimated. Conventional acute/chronic toxicity tests were conducted using Japanese medaka (Oryzias latipes), daphnia (Daphnia magna), and green algae (Pseudokirchneriella subcapitata) for n-butylparaben, i-butylparaben, and benzylparaben. Medaka vitellogenin assays were also conducted for the three compounds and DNA microarray analysis was carried out to examine the effects of benzylparaben on gene expression. The plasma vitellogenin concentration of male medaka increased for concentrations of 200, 100, and 100 microg L(-1) n-butylparaben, i butylparaben, and benzylparaben for 14 days, respectively, while the expression levels of genes encoding proteins such as p53, cytochrome P450 3A40, and choriogenin-L increased for concentrations higher than 4 microg L(-1) of benzylparaben. Furthermore, the predicted no-effect concentration (PNEC) was calculated using the lethal or effect concentration 50 (LC50 or EC50) values and no-effect concentrations (NOECs) obtained in the toxicity tests for these compounds. The maximum concentrations found in the aquatic environment or sewage effluent (MEC eff) were used to carry out preliminary environmental risk assessment. The calculated MEC/PNEC ratio suggests the necessity of further study such as a more detailed large-scale monitoring and chronic toxicity tests including reproduction inhibition and endocrine disruption. PMID- 18382417 TI - Suppression of formation of dioxins in combustion gas of municipal waste incinerators by spray water injection. AB - Dioxins in the combustion gas of municipal solid waste incinerators (MSWIs) are resynthesized when the combustion gas passes from the outlet exaust gas boiler to the outlet gas duct. The objective of the study was to estimate if the suppression of the formation of dioxins depends on the inlet gas temperature and diameter and/or temperature of droplet spray water using an actual incinerator operation data. The dioxin formation and/or the quenching temperature is revealed using the Altwicker theory equation with the information of inlet gas temperature and droplet spray water. The evaporation rate of a spray water droplet also can be estimated using the Mizutani theory. The highest dioxin formation was found at 350 degrees C; thereafter, it decreased quickly. When an area of 500 microm for droplet-formed dioxins is defined as 100%, the values of formed dioxins for 400, 300, 200 and 100 microm droplet areas are estimated as 71, 41, 25 and 18%, respectively. It is revealed that the smaller size of droplet spray water and lower inlet gas temperature enable the decrease in dioxin formation. The decreased dioxin formation and/or the lower quenching temperature is revealed using the Altwicker theory equation with the information of inlet gas temperature and droplet spray water size. PMID- 18382418 TI - Preliminary ecological risk assessment of butylparaben and benzylparaben -2. Fate and partitioning in aquatic environments. AB - Butylparaben and benzylparaben, used as preservatives mainly in cosmetic products, have recently been shown to be weakly estrogenic. Batch sunlight photolysis and river water biodegradation experiments were conducted to determine the persistence of these compounds in aquatic environments. As a result, benzylparaben was found to be moderately photodegradable whereas both n butylparaben and i-butylparaben were highly stable against sunlight. Both benzylparaben and butylparabens were relatively biodegradable in the river water but the degradability was dependent on the sampling site and time. Batch sorption experiments were also conducted to determine the coefficients of sorption into river sediments and a model soil sample. The determined coefficients were slightly higher for benzylparaben than the two butylparabens and comparable to that of the natural estrogen 17beta-estradiol. The coefficients were also higher for sediment/soil with a higher organic content and the organic-carbon-based sorption coefficient (log K oc) shows a moderate linear correlation with the octanol-water partition coefficient (log K ow). These results suggest that hydrophobic interaction plays a predominant role in sorption at neutral pH. PMID- 18382420 TI - Does tertiary Gleason score 5 have prognostic significance in men with prostate cancer? PMID- 18382419 TI - Mechanisms of disease: molecular genetics of arrhythmogenic right ventricular dysplasia/cardiomyopathy. AB - Arrhythmogenic right ventricular dysplasia/cardiomyopathy is an inherited cardiomyopathy estimated to affect approximately 1 in 5,000 individuals. Cardinal manifestations include right ventricular enlargement and dysfunction, fibrofatty replacement of myocytes in the right ventricle, characteristic electrocardiographic abnormalities, and ventricular arrhythmia most commonly arising from the right ventricle. The disease is frequently familial and typically involves autosomal dominant transmission with low penetrance and variable expressivity. Approximately 50% of symptomatic individuals harbor a mutation in one of the five major components of the cardiac desmosome. Nevertheless, other genetic modifiers and environmental factors complicate the clinical management of mutation carriers as well as counseling of their relatives. This Review summarizes the known genetic mutations associated with arrhythmogenic right ventricular dysplasia/cardiomyopathy, describes possible origins of recurrent mutations, presents theories on the pathogenesis of disease following a mutation, and discusses the current issues surrounding clinical use of genetic analysis in the assessment of individuals with this condition. PMID- 18382421 TI - Is there a relationship between urinary tract infection, vesicoureteral reflux and renal damage in children? PMID- 18382422 TI - Does primary etiology affect outcomes in adults with childhood-onset growth hormone deficiency? PMID- 18382423 TI - Differentiating between body fat and lean mass-how should we measure obesity? PMID- 18382424 TI - Is sitaxsentan a good therapeutic option for pulmonary arterial hypertension associated with connective tissue disease? AB - Agents that indiscriminately block endothelin receptors have been shown to cause moderate improvements in the outcomes of patients who have pulmonary arterial hypertension (PAH) as a complication of connective tissue disease (CTD). Girgis et al. investigated the effects of sitaxsentan, a selective endothelin receptor type A antagonist, in patients with CTD and comcomitant PAH. After 12 weeks of treatment, patients who received sitaxsentan showed more improvement from baseline compared with those receiving placebo in terms of exercise capacity, hemodynamics and physical-health-related quality of life. At the end of the extension study (median total follow-up 26 weeks), 16 of the 41 patients with CTD had an improvement of at least one New York Heart Association functional class compared with at the start of sitaxsentan therapy. The effects of sitaxsentan observed in the CTD group were comparable to those seen in the idiopathic PAH group. The authors concluded that treatment with sitaxsentan might be beneficial in patients with PAH associated with CTD. PMID- 18382425 TI - Single-step doxorubicin-selected cancer cells overexpress the ABCG2 drug transporter through epigenetic changes. AB - Understanding the mechanisms of multidrug resistance (MDR) could improve clinical drug efficacy. Multidrug resistance is associated with ATP binding cassette (ABC) transporters, but the factors that regulate their expression at clinically relevant drug concentrations are poorly understood. We report that a single-step selection with low doses of anti-cancer agents, similar to concentrations reported in vivo, induces MDR that is mediated exclusively by ABCG2. We selected breast, ovarian and colon cancer cells (MCF-7, IGROV-1 and S-1) after exposure to 14 or 21 nM doxorubicin for only 10 days. We found that these cells overexpress ABCG2 at the mRNA and protein levels. RNA interference analysis confirmed that ABCG2 confers drug resistance. Furthermore, ABCG2 upregulation was facilitated by histone hyperacetylation due to weaker histone deacetylase 1-promoter association, indicating that these epigenetic changes elicit changes in ABCG2 gene expression. These studies indicate that the MDR phenotype arises following low-dose, single-step exposure to doxorubicin, and further suggest that ABCG2 may mediate early stages of MDR development. This is the first report to our knowledge of single-step, low-dose selection leading to overexpression of ABCG2 by epigenetic changes in multiple cancer cell lines. PMID- 18382426 TI - Animal foods, protein, calcium and prostate cancer risk: the European Prospective Investigation into Cancer and Nutrition. AB - We examined consumption of animal foods, protein and calcium in relation to risk of prostate cancer among 142 251 men in the European Prospective Investigation into Cancer and Nutrition. Associations were examined using Cox regression, stratified by recruitment centre and adjusted for height, weight, education, marital status and energy intake. After an average of 8.7 years of follow-up, there were 2727 incident cases of prostate cancer, of which 1131 were known to be localised and 541 advanced-stage disease. A high intake of dairy protein was associated with an increased risk, with a hazard ratio for the top versus the bottom fifth of intake of 1.22 (95% confidence interval (CI): 1.07-1.41, P(trend)=0.02). After calibration to allow for measurement error, we estimated that a 35-g day(-1) increase in consumption of dairy protein was associated with an increase in the risk of prostate cancer of 32% (95% CI: 1-72%, P(trend)=0.04). Calcium from dairy products was also positively associated with risk, but not calcium from other foods. The results support the hypothesis that a high intake of protein or calcium from dairy products may increase the risk for prostate cancer. PMID- 18382427 TI - A phase II study of sequential neoadjuvant gemcitabine plus doxorubicin followed by gemcitabine plus cisplatin in patients with operable breast cancer: prediction of response using molecular profiling. AB - This study examined the pathological complete response (pCR) rate and safety of sequential gemcitabine-based combinations in breast cancer. We also examined gene expression profiles from tumour biopsies to identify biomarkers predictive of response. Indian women with large or locally advanced breast cancer received 4 cycles of gemcitabine 1200 mg m(-2) plus doxorubicin 60 mg m(-2) (Gem+Dox), then 4 cycles of gemcitabine 1000 mg m(-2) plus cisplatin 70 mg m(-2) (Gem+Cis), and surgery. Three alternate dosing sequences were used during cycle 1 to examine dynamic changes in molecular profiles. Of 65 women treated, 13 (24.5% of 53 patients with surgery) had a pCR and 22 (33.8%) had a complete clinical response. Patients administered Gem d1, 8 and Dox d2 in cycle 1 (20 of 65) reported more toxicities, with G3/4 neutropenic infection/febrile neutropenia (7 of 20) as the most common cycle-1 event. Four drug-related deaths occurred. In 46 of 65 patients, 10-fold cross validated supervised analyses identified gene expression patterns that predicted with >or=73% accuracy (1) clinical complete response after eight cycles, (2) overall clinical complete response, and (3) pCR. This regimen shows strong activity. Patients receiving Gem d1, 8 and Dox d2 experienced unacceptable toxicity, whereas patients on other sequences had manageable safety profiles. Gene expression patterns may predict benefit from gemcitabine-containing neoadjuvant therapy. PMID- 18382428 TI - Acquisition of biologically relevant gene expression data by Affymetrix microarray analysis of archival formalin-fixed paraffin-embedded tumours. AB - Robust protocols for microarray gene expression profiling of archival formalin fixed paraffin-embedded tissue (FFPET) are needed to facilitate research when availability of fresh-frozen tissue is limited. Recent reports attest to the feasibility of this approach, but the clinical value of these data is poorly understood. We employed state-of-the-art RNA extraction and Affymetrix microarray technology to examine 34 archival FFPET primary extremity soft tissue sarcomas. Nineteen arrays met stringent QC criteria and were used to model prognostic signatures for metastatic recurrence. Arrays from two paired frozen and FFPET samples were compared: although FFPET sensitivity was low ( approximately 50%), high specificity (95%) and positive predictive value (92%) suggest that transcript detection is reliable. Good agreement between arrays and real time (RT)-PCR was confirmed, especially for abundant transcripts, and RT-PCR validated the regulation pattern for 19 of 24 candidate genes (overall R(2)=0.4662). RT-PCR and immunohistochemistry on independent cases validated prognostic significance for several genes including RECQL4, FRRS1, CFH and MET - whose combined expression carried greater prognostic value than tumour grade - and cmet and TRKB proteins. These molecules warrant further evaluation in larger series. Reliable clinically relevant data can be obtained from archival FFPET, but protocol amendments are needed to improve the sensitivity and broad application of this approach. PMID- 18382429 TI - Metastatic colorectal cancer: irinotecan plus infusional, bolus or oral fluoropyrimidines as first-line treatment. PMID- 18382430 TI - Is the prevalence of colorectal neoplasm higher in patients with coronary artery disease? PMID- 18382431 TI - Which patients with prostate cancer benefit from immediate postoperative radiotherapy? PMID- 18382432 TI - Mechanisms of disease: the role of gastrointestinal hormones in appetite and obesity. AB - The obesity epidemic is fast becoming one of the leading causes of mortality and morbidity worldwide. Over the past 30 years, gastrointestinal hormones have been increasingly understood to have an important role as regulators of appetite and energy balance in obese individuals. The levels of these hormones are modulated by bariatric surgery, and understanding how they are affected by such procedures can contribute to our comprehension of the underlying mechanisms by which these hormones affect obesity and its treatment. In this Review, we consider several gastrointestinal hormones that can contribute to obesity by modulating the activity of the gut-brain axis, and examine their specific effects on appetite, hunger and energy balance. Better understanding of the mechanisms by which these peptides exert their effects may enable the development of improved weight-loss medications and new treatments for obesity. PMID- 18382433 TI - Is CT colonography superior to colonoscopy for the detection of advanced neoplasia? PMID- 18382434 TI - Is virtual reality simulation an effective training method in surgery? PMID- 18382435 TI - Drug insight: antiangiogenic therapies for gastrointestinal cancers--focus on monoclonal antibodies. AB - Tumor angiogenesis is strongly induced by vascular endothelial growth factor (VEGF), which is overexpressed in most human gastrointestinal cancers. VEGF overexpression is known to be associated with poor prognosis and survival in patients with various solid tumors. The humanized monoclonal anti-VEGF antibody bevacizumab (Avastin, Genentech Inc., South San Francisco, CA) is a prototypic antiangiogenic compound, and has proven therapeutic benefit combined with conventional chemotherapy-namely, significantly improved progression-free survival in patients with metastatic colorectal cancer. Bevacizumab is the only anti-VEGF antibody that has been approved by the FDA and the European Medicines Agency for the treatment of metastatic colorectal cancer. Several ongoing clinical studies are evaluating the potential of bevacizumab therapy for other gastrointestinal cancers, in combination with chemotherapy, other targeted therapies and/or radiation. Soluble chimeric receptors, tyrosine kinase inhibitors, and monoclonal antibodies against VEGF and molecular targets in the integrin and Delta-like protein 4-Notch pathways are being developed. As tumors acquire resistance to anti-VEGF therapy, further development of antiangiogenic and vascular targets and therapy is warranted. PMID- 18382436 TI - Should hepatitis C virus infection be a contraindication to renal transplantation? PMID- 18382437 TI - Technology insight: imaging neurodegeneration in Parkinson's disease. AB - Currently, the clinical diagnosis of Parkinson's disease (PD) can be problematic, particularly at the early stages of the disease when the full spectrum of symptoms and signs might not yet be manifest. In addition, the mechanisms that underlie the nonmotor complications of PD, such as dementia and depression, are poorly understood, despite the fact that these symptoms largely determine the patient's quality of life at the end stage of the disease. This article reviews the latest advances in structural and functional imaging that have provided important insights into the structural, pathophysiological and pharmacological changes associated with PD. The contribution of inflammatory processes to the pathology of PD is discussed, as are the various possible mechanisms that lead to coexistent dementia and depression. PMID- 18382438 TI - Determination of T-cell fate by dendritic cells. AB - Dendritic cells (DC) are professional antigen-presenting cells with a unique T cell stimulatory aptitude that play a crucial role in the instruction of adaptive immune responses upon infection. By controlling the initiation of a diverse set of effector functions, which are suitable for the elimination of a wide range of pathogens, DCs form the pivotal link between the innate and the adaptive immune system. The innate pattern recognition pathways that trigger DC activation are central for skewing of the adaptive immune responses that are subsequently induced. Thus innate activation not only precedes adaptive immune activation, it also controls it and tailors the effector functions to the requirements of the infection. The adaptive immune response has to match the nature of the infection, but this does not only concern the type of pathogen, it is also affected by the localization of the infection. Tissue homeostasis has to be ensured and thus tissue-derived environmental factors influence the functional activity of activated DCs and thereby contribute to shaping of the immune response. Adaptive immune responses are vital for the elimination of pathogens, have the potential to attack tumor cells and play a detrimental role during transplant rejection and in a variety of autoimmune diseases. Better understanding of the mechanisms that control the induction of different T-cell effector functions will enable the development of strategies to manipulate the immune system in the context of vaccination, tumor immunotherapy, transplantation and autoimmunity. PMID- 18382439 TI - T-cell migration: Kruppeled T cells move again. PMID- 18382440 TI - KDIGO clinical practice guidelines for the prevention, diagnosis, evaluation, and treatment of hepatitis C in chronic kidney disease. PMID- 18382459 TI - Surveillance of cirrhosis for hepatocellular carcinoma: a cost-utility analysis. AB - Using a decision-analytic model, we evaluated the effectiveness and cost effectiveness of surveillance for hepatocellular carcinoma (HCC) in individuals with cirrhosis. Separate cohorts with cirrhosis due to alcoholic liver disease, hepatitis B and hepatitis C were simulated. Results were also combined to approximate a mixed aetiology population. Comparisons were made between a variety of surveillance algorithms using alpha-foetoprotein (AFP) assay and/or ultrasound at 6- and 12-monthly intervals. Parameter estimates were obtained from comprehensive literature reviews. Uncertainty was explored using one-way and probabilistic sensitivity analyses. In the mixed aetiology cohort, 6-monthly AFP+ultrasound was predicted to be the most effective strategy. The model estimates that, compared with no surveillance, this strategy may triple the number of people with operable tumours at diagnosis and almost halve the number of people who die from HCC. The cheapest strategy employed triage with annual AFP (incremental cost-effectiveness ratio (ICER): 20,700 pounds per quality-adjusted life-year (QALY) gained). At a willingness-to-pay threshold of 30,000 pounds per QALY the most cost-effective strategy used triage with 6-monthly AFP (ICER: 27,600 pounds per QALY gained). The addition of ultrasound to this strategy increased the ICER to 60,100 pounds per QALY gained. Surveillance appears most cost-effective in individuals with hepatitis B-related cirrhosis, potentially due to younger age at diagnosis of cirrhosis. Our results suggest that, in a UK NHS context, surveillance of individuals with cirrhosis for HCC should be considered effective and cost-effective. The economic efficiency of different surveillance strategies is predicted to vary markedly according to cirrhosis aetiology. PMID- 18382460 TI - Assessment of 1183 screen-detected, category 3B, circumscribed masses by cytology and core biopsy with long-term follow up data. AB - Discrete masses are commonly detected during mammographic screening and most such lesions are benign. For lesions without pathognomonically benign imaging features that are still regarded likely to be non-malignant (Tabar grade 3) reliable biopsy results would be a clinically useful alternative to mammographic surveillance. Appropriate institutional guidelines for ethical research were followed. Between Jan 1996--Dec 2005 grade 3B discrete masses detected in the setting of a large, population based, breast cancer screening programme are included. Patient demographics, fine needle aspiration biopsy (FNAB), core and surgical biopsy results are tabulated. The final pathology of excised lesions was obtained. Information regarding interval cancers was obtained from the State Cancer Registry records and also through long term follow-up of clients in subsequent rounds of screening. A total of 1183 lesions, mean diameter of 13.3 mm (+/-8.3 mm) and mean client age of 55.1 years (+/-8.8 years) are included. After diagnostic work up, 98 lesions (8.3%) were malignant, 1083 were non-malignant and a final histologic diagnosis was not established in two lesions. In the 27 months after assessment, no interval cancers were attributable to these lesions and during a mean follow up of 54.5 months, available in 84.9% of eligible women, only one cancer has developed in the same quadrant as the original lesion, although the two processes are believed to be unrelated. FNAB performed in 1149 cases was definitive in 80.5% cases (882 benign, 43 malignant) with a negative predictive value (NPV) of 99.8% (880 of 882) and a positive predictive value (PPV) of 95.2% (40 of 42, both intraductal papillomas). Core biopsy was performed in 178 lesions, mostly for indefinite cytology. Core biopsy was definitive in 79.8% cases (57% benign 22% malignant) with a PPV of 100% and NPV of 99.0%. In experienced hands FNAB is an accurate first line diagnostic modality for the assessment of 3B screen-detected discrete masses, providing definitive results in 80.5% of cases. When used as a second line modality, core biopsy had a similarly high rate of definitive diagnosis at 79.8%. The stepwise approach to the use of FNAB and core biopsy would reduce substantially the proportion of cases requiring surgical diagnostic biopsy. Given the low probability of malignancy and the imperative to limit the morbidity associated with cancer screening, the demonstration of the reliability of FNAB as a minimally invasive but highly accurate test for this particular subset of screen-detected lesions has significant clinical utility. PMID- 18382461 TI - Parity and risk of stomach cancer by sub-site: a national Swedish study. AB - We investigated stomach cancer risk by anatomic sub-site in relation to parity, as a marker for higher exposure to sex hormones, in a case-control study, nested within a cohort of 2,406,439 Swedish women born in 1925 or later and followed from 1970 or age 30 until emigration, death, any cancer diagnosis, or through 2004, whichever occurred first. We identified 286 cardia and 2498 non-cardia stomach cancer cases with five matched controls for each case. Cross-linkage with the Multi-Generation Register provided information about reproductive history. Using conditional logistic regression models for estimating odds ratios (ORs) and corresponding 95% confidence intervals (CIs), adjusted for education level and occupation, we found no association between any aspect of parity and non-cardia stomach cancer (OR=1.01, 95% CI 0.89-1.15, comparing parous with nulliparous women). However, a 30% risk reduction for postmenopausal cardia cancer (OR=0.7, 95% CI 0.4-1.0) was noted among parous relative to nulliparous women and the risk for premenopausal cardia cancer fell with increasing number of children (P for trend=0.04). Our results indicate that exposure to female sex hormones does not protect against non-cardia stomach cancer and does not explain male predominance. The observed moderate inverse relationship between parity and cardia cancer may be mediated by non-hormonal factors and warrants further study. PMID- 18382462 TI - Developmental axon pruning mediated by BDNF-p75NTR-dependent axon degeneration. AB - The mechanisms that regulate the pruning of mammalian axons are just now being elucidated. Here, we describe a mechanism by which, during developmental sympathetic axon competition, winning axons secrete brain-derived neurotrophic factor (BDNF) in an activity-dependent fashion, which binds to the p75 neurotrophin receptor (p75NTR) on losing axons to cause their degeneration and, ultimately, axon pruning. Specifically, we found that pruning of rat and mouse sympathetic axons that project to the eye requires both activity-dependent BDNF and p75NTR. p75NTR and BDNF are also essential for activity-dependent axon pruning in culture, where they mediate pruning by directly causing axon degeneration. p75NTR, which is enriched in losing axons, causes axonal degeneration by suppressing TrkA-mediated signaling that is essential for axonal maintenance. These data provide a mechanism that explains how active axons can eliminate less-active, competing axons during developmental pruning by directly promoting p75NTR-mediated axonal degeneration. PMID- 18382463 TI - Defining drug disposition determinants: a pharmacogenetic-pharmacokinetic strategy. AB - In preclinical and early clinical drug development, information about the factors influencing drug disposition is used to predict drug interaction potential, estimate and understand population pharmacokinetic variability, and select doses for clinical trials. However, both in vitro drug metabolism studies and pharmacogenetic association studies on human pharmacokinetic parameters have focused on a limited subset of the proteins involved in drug disposition. Furthermore, there has been a one-way information flow, solely using results of in vitro studies to select candidate genes for pharmacogenetic studies. Here, we propose a two-way pharmacogenetic-pharmacokinetic strategy that exploits the dramatic recent expansion in knowledge of functional genetic variation in proteins that influence drug disposition, and discuss how it could improve drug development. PMID- 18382464 TI - Structural diversity of G protein-coupled receptors and significance for drug discovery. AB - G protein-coupled receptors (GPCRs) are the largest family of membrane-bound receptors and also the targets of many drugs. Understanding of the functional significance of the wide structural diversity of GPCRs has been aided considerably in recent years by the sequencing of the human genome and by structural studies, and has important implications for the future therapeutic potential of targeting this receptor family. This article aims to provide a comprehensive overview of the five main human GPCR families--Rhodopsin, Secretin, Adhesion, Glutamate and Frizzled/Taste2--with a focus on gene repertoire, general ligand preference, common and unique structural features, and the potential for future drug discovery. PMID- 18382467 TI - Hippocampal and perirhinal functions in recognition memory. PMID- 18382468 TI - Metabolic origin of BOLD signal fluctuations in the absence of stimuli. AB - Blood oxygen level-dependent (BOLD) functional magnetic resonance imaging studies have shown the existence of ongoing blood flow fluctuations in the absence of stimuli. Although this so-called 'resting-state activity' appears to be correlated across brain regions with apparent functional relationship, its origin might be predominantly vascular and not directly representing neuronal signaling. To investigate this, we simultaneously measured BOLD and perfusion signals on healthy human subjects (n=11) and used their ratio (BOLD/perfusion ratio or BPR) as an indicator of metabolic demand. BPR during rest and sleep was compared with that during a visual task (VT) and a breath-holding task (BH), which are challenges with substantial and little metabolic involvement, respectively. Within the visual cortex, BPR was 3.76+/-1.23 during BH, which was significantly higher than during the VT (1.76+/-0.27) and rest (1.56+/-0.41). Meanwhile, BPR values during VT and rest were not significantly different, suggesting a similar metabolic involvement. Eight subjects showed stage 1 and 2 sleep, during which temporally correlated BOLD and perfusion activity continued. In these subjects, there was no significant difference in BPR between the sleep and waking conditions (1.79+/-0.54 and 1.66+/-0.67, respectively), but both were lower than the BPR during BH. These data suggest that resting-state activity, at least in part, represents a metabolic process. PMID- 18382465 TI - GABA(A) receptor trafficking and its role in the dynamic modulation of neuronal inhibition. AB - GABA (gamma-aminobutyric acid) type A receptors (GABA(A)Rs) mediate most fast synaptic inhibition in the mammalian brain, controlling activity at both the network and the cellular levels. The diverse functions of GABA in the CNS are matched not just by the heterogeneity of GABA(A)Rs, but also by the complex trafficking mechanisms and protein-protein interactions that generate and maintain an appropriate receptor cell-surface localization. In this Review, we discuss recent progress in our understanding of the dynamic regulation of GABA(A)R composition, trafficking to and from the neuronal surface, and lateral movement of receptors between synaptic and extrasynaptic locations. Finally, we highlight a number of neurological disorders, including epilepsy and schizophrenia, in which alterations in GABA(A)R trafficking occur. PMID- 18382469 TI - Additive effects of statin and dipyridamole on cerebral blood flow and stroke protection. AB - Recent studies suggest that dipyridamole (DP) may exert stroke protective effects beyond platelet inhibition. The purpose of this study is to determine whether statin and DP could enhance stroke protection through nitric oxide (NO)-dependent vascular effects. Mice were pretreated with DP (10 to 60 mg/kg, q 12 h, 3 days) alone or in combination with a statin (simvastatin; 0.1 to 20 mg/kg per day, 14 days) before transient intraluminal middle cerebral artery occlusion. Although simvastatin (1 mg/kg per day, 14 days) increased endothelial NO synthase (eNOS) activity by 25% and DP (30 mg/kg, q12 h, 3 days) increased aortic cGMP levels by 55%, neither statin nor DP alone, at these subtherapeutic doses, increased absolute cerebral blood flow (CBF) or conferred stroke protection. However, the combination of subtherapeutic doses of simvastatin and DP increased CBF by 50%, decreased stroke volume by 54%, and improved neurologic motor deficits, all of which were absent in eNOS-deficient mice. In contrast, treatment with aspirin (10 mg/kg per day, 3 days) did not augment the neuroprotective effects of DP and/or simvastatin. These findings indicate that statin and DP exert additive NO dependent vascular effects and suggest that the combination of statin and DP has greater benefits in stroke protection than statin alone through vascular protection. PMID- 18382470 TI - Gene expression in peripheral blood differs after cardioembolic compared with large-vessel atherosclerotic stroke: biomarkers for the etiology of ischemic stroke. AB - There are no biomarkers that differentiate cardioembolic from large-vessel atherosclerotic stroke, although the treatments differ for each and approximately 30% of strokes and transient ischemic attacks have undetermined etiologies using current clinical criteria. We aimed to define gene expression profiles in blood that differentiate cardioembolic from large-vessel atherosclerotic stroke. Peripheral blood samples were obtained from healthy controls and acute ischemic stroke patients (<3, 5, and 24 h). RNA was purified, labeled, and applied to Affymetrix Human U133 Plus 2.0 Arrays. Expression profiles in the blood of cardioembolic stroke patients are distinctive from those of large-vessel atherosclerotic stroke patients. Seventy-seven genes differ at least 1.5-fold between them, and a minimum number of 23 genes differentiate the two types of stroke with at least 95.2% specificity and 95.2% sensitivity for each. Genes regulated in large-vessel atherosclerotic stroke are expressed in platelets and monocytes and modulate hemostasis. Genes regulated in cardioembolic stroke are expressed in neutrophils and modulate immune responses to infectious stimuli. This new method can be used to predict whether a stroke of unknown etiology was because of cardioembolism or large-vessel atherosclerosis that would lead to different therapy. These results have wide ranging implications for similar disorders. PMID- 18382471 TI - Acute functional recovery of cerebral blood flow after forebrain ischemia in rat. AB - After complete cerebral ischemia, the postischemic blood flow response to functional activation is severely attenuated for several hours. However, little is known about the spatial and temporal extent of the blood flow response in the acute postischemic period after incomplete cerebral ischemia. To investigate the relative cerebral blood flow (rCBF) response in the somatosensory cortex of rat to controlled vibrissae stimulation after transient incomplete ischemia (15-min bilateral common carotid artery occlusion+hypotension), we employed laser speckle imaging combined with statistical parametric mapping. We found that the ischemic insult had a significant impact on the baseline blood flow (P<0.005) and the activation area in response to functional stimulation was significantly reduced after ischemia (P<0.005). The maximum rCBF response in the activation area determined from the statistical analysis did not change significantly up to 3 h after ischemia (P>0.1). However, the time when rCBF response reached its maximum was significantly delayed (P<0.0001) from 2.4+/-0.2 secs before ischemia to 3.6+/ 0.1 secs at 20 mins into reperfusion (P<0.001); the delay was reduced gradually to 2.9+/-0.2 secs after 3 h, which was still significantly greater than that observed before the insult (P=0.04). PMID- 18382472 TI - The epithelial membrane protein 1 is a novel tight junction protein of the blood brain barrier. AB - In the central nervous system, a constant microenvironment required for neuronal cell activity is maintained by the blood-brain barrier (BBB). The BBB is formed by the brain microvascular endothelial cells (BMEC), which are sealed by tight junctions (TJ). To identify genes that are differentially expressed in BMEC compared with peripheral endothelial cells, we constructed a subtractive cDNA library from porcine BMEC (pBMEC) and aortic endothelial cells (AOEC). Screening the library for differentially expressed genes yielded 26 BMEC-specific transcripts, such as solute carrier family 35 member F2 (SLC35F2), ADP ribosylation factor-like 5B (ARL5B), TSC22 domain family member 1 (TSC22D1), integral membrane protein 2A (ITM2A), and epithelial membrane protein 1 (EMP1). In this study, we show that EMP1 transcript is enriched in pBMEC compared with brain tissue and that EMP1 protein colocalizes with the TJ protein occludin in mouse BMEC by coimmunoprecipitation and in rat brain vessels by immunohistochemistry. Epithelial membrane protein 1 expression was transiently induced in laser-capture microdissected rat brain vessels after a 20-min global cerebral ischemia, in parallel with the loss of occludin immunoreactivity. The study identifies EMP1 as a novel TJ-associated protein of the BBB and suggests its potential role in the regulation of the BBB function in cerebral ischemia. PMID- 18382473 TI - Meiotic hybridogenesis in triploid Misgurnus loach derived from a clonal lineage. AB - Triploid loaches Misgurnus anguillicaudatus are derived from unreduced diploid gametes produced by an asexual clonal lineage that normally undergoes gynogenetic reproduction. Here, we have investigated the reproductive system of two types of triploids: the first type carried maternally inherited clonal diploid genomes and a paternally inherited haploid genome from the same population; the second type had the same clonal diploid genomes but a haploid genome from another, genetically divergent population. The germinal vesicles of oocytes from triploid females (3n=75) contained only 25 bivalents, that is, 50 chromosomes. Flow cytometry revealed that the majority of the progeny resulting from fertilization of eggs from triploid females with normal haploid sperm were diploid. This indicates that triploid females mainly produced haploid eggs. Microsatellite analyses of the diploid progeny of triploid females showed that one allele of the clonal genotype was not transmitted to haploid eggs. Moreover, the identity of the eliminated allele differed between the two types of triploids. Our results demonstrate that there is preferential pairing of homologous chromosomes as well as the elimination of unmatched chromosomes in the course of haploid egg formation, that is, meiotic hybridogenesis. Two distinct genomes in the clone suggest its hybrid origin. PMID- 18382474 TI - Interleukin 18 receptor 1 gene polymorphisms are associated with asthma. AB - The interleukin 18 receptor (IL18R1) gene is a strong candidate gene for asthma. It has been implicated in the pathophysiology of asthma and maps to an asthma susceptibility locus on chromosome 2q12. The possibility of association between polymorphisms in IL18R1 and asthma was examined by genotyping seven SNPs in 294, 342 and 100 families from Denmark, United Kingdom and Norway and conducting family-based association analyses for asthma, atopic asthma and bronchial hyper reactivity (BHR) phenotypes. Three SNPs in IL18R1 were associated with asthma (0.01131 < or = P < or = 0.01377), five with atopic asthma (0.00066 < or = P < or = 0.00405) and two with BHR (0.01450 < or = P < or = 0.03203) in the Danish population; two SNPs were associated with atopic asthma (0.00397 < or = P < or = 0.01481) and four with BHR (0.00435 < or = P < or = 0.03544) in the UK population; four SNPs showed associations with asthma (0.00015 < or = P < or = 0.03062), two with atopic asthma (0.01269 < or = P < or = 0.04042) and three with BHR (0.00259 < or = P < or = 0.01401) in the Norwegian population; five SNPs showed associations with asthma (0.00005 < or = P < or = 0.03744), five with atopic asthma (0.00001 < or = P < or = 0.04491) and three with BHR (0.03568 < or = P < or = 0.04778) in the combined population. Three intronic SNPs (rs1420099, rs1362348 and rs1974675) showed replicated association for at least one asthma related phenotype. These results demonstrate significant association between polymorphisms in IL18R1 and asthma. PMID- 18382475 TI - Identification of a founder mutation in TPM3 in nemaline myopathy patients of Turkish origin. AB - To date, six genes are known to cause nemaline (rod) myopathy (NM), a rare congenital neuromuscular disorder. In an attempt to find a seventh gene, we performed linkage and subsequent sequence analyses in 12 Turkish families with recessive NM. We found homozygosity in two of the families at 1q12-21.2, a region encompassing the gamma-tropomyosin gene (TPM3) encoding slow skeletal muscle alpha-tropomyosin, a known NM gene. Sequencing revealed homozygous deletion of the first nucleotide of the last exon, c.913delA of TPM3 in both families. The mutation removes the last nucleotide before the stop codon, causing a frameshift and readthrough across the termination signal. The encoded alphaTm(slow) protein is predicted to be 73 amino acids longer than normal, and the extension to the protein is hypothesised to be unable to form a coiled coil. The resulting tropomyosin protein may therefore be non-functional. The affected children in both families were homozygous for the mutation, while the healthy parents were mutation carriers. Both of the patients in Family 1 had the severe form of NM, and also an unusual chest deformity. The affected children in Family 2 had the intermediate form of NM. Muscle biopsies showed type 1 (slow) fibres to be markedly smaller than type 2 (fast) fibres. Previously, there had been five reports, only, of NM caused by mutations in TPM3. The mutation reported here is the first deletion to be identified in TPM3, and it is likely to be a founder mutation in the Turkish population. PMID- 18382476 TI - Cox proportional hazards models have more statistical power than logistic regression models in cross-sectional genetic association studies. AB - Cross-sectional genetic association studies can be analyzed using Cox proportional hazards models with age as time scale, if age at onset of disease is known for the cases and age at data collection is known for the controls. We assessed to what degree and under what conditions Cox proportional hazards models have more statistical power than logistic regression models in cross-sectional genetic association analyses. Analyses were conducted in an empirical study on the association of 65 polymorphisms and risk of coronary heart disease among 2400 familial hypercholesterolemia patients, and in a simulation study that considered various combinations of sample size, genotype frequency, and strength of association between the genotype and coronary heart disease. We applied Cox proportional hazards models and logistic regression models, and compared effect estimates (hazard ratios and odds ratios) and statistical power. In the empirical study, Cox proportional hazards models generally showed lower P-values for polymorphisms than logistic regression models. In the simulation study, Cox proportional hazards models had higher statistical power in all scenarios. Absolute differences in power did depend on the effect estimate, genotype frequency and sample size, and were most prominent for genotypes with minor effects. For example, when the genotype frequency was 30% in a sample with size n=2000 individuals, the absolute differences were the largest for effect estimates between 1.1 and 1.5. In conclusion, Cox proportional hazards models can increase statistical power in cross-sectional genetic association studies, especially in the range of effect estimates that are expected for genetic associations in common diseases. PMID- 18382477 TI - Catechol O-methyl transferase and dopamine D2 receptor gene polymorphisms: evidence of positive heterosis and gene-gene interaction on working memory functioning. AB - The COMT Val(108/158)Met polymorphism has been extensively studied in relation to individual differences in working memory (WM) performance. The present study tested the association of the COMT Val(108/158)Met polymorphism with WM performance in two independent family-based Dutch samples: 371 children (mean age 12.4 years) and 391 adults (mean age 36.2 years). A significant association was found between the COMT polymorphism and WM scores in the combined adult and young cohorts. The association reflected positive heterosis such that the Met/Met and Val/Val homozygotes did not perform as well as the Met/Val heterozygotes on the WM tasks. A secondary analysis was conducted in which a DRD2-tagging SNP (rs2075654) was tested for an interactive effect with the COMT polymorphism on WM performance. A significant interactive effect of the DRD2 and COMT genes was found such that heterosis was present only in the DRD2 genotype that has been linked to lower receptor density. Our results support previous findings that WM performance needs an optimal level of dopamine signaling within the PFC. This optimum level depends on enzymatic activity controlling dopamine level as well as dopamine receptor sensitivity, both of which may differ as a function of age and genotype. We conclude that the effects of a single polymorphism in a dopaminergic gene on a well-defined cognitive trait may easily remain hidden if the interaction with age and other genes in the pathway are not taken into account. PMID- 18382478 TI - The Public Population Project in Genomics (P3G): a proof of concept? PMID- 18382479 TI - Establishment of a biobank and pharmacogenetics database of African populations. PMID- 18382480 TI - Rheumatology in the current era: the challenge of success. PMID- 18382481 TI - Far field of gratings with rough strips. AB - In this work, we analyze the far-field pattern produced by a grating made of strips with two different random roughness levels. The efficiency and shape of the diffraction orders is obtained, which are shown to depend on the statistical properties of roughness. We assume for the calculations that the grating can be used in a mobile mechanical system. A preliminary experimental approach which partially corroborates the theoretical results is also performed. PMID- 18382482 TI - Efficiency of linear-to-circular polarization conversion for light reflection at the principal angle by a dielectric-conductor interface. AB - The efficiency eta(LC) of linear-to-circular polarization conversion when light is reflected at a dielectric-conductor interface is determined as a function of the principal angle phi and principal azimuth psi . Constant-eta(LC) contours are presented in the phi , psi plane for values of eta(LC) from 0.5 to 1.0 in steps of 0.05, and the corresponding contours in the complex plane of the relative dielectric function are also determined. As specific examples, efficiencies > or = 88% are obtained for light reflection by a Ag mirror in the visible and near-IR (400-1200 nm) spectral range, and > or =40% for the reflection of extreme ultraviolet (EUV) and soft x-ray radiation by a SiC mirror in the 60-120 nm wavelength range. PMID- 18382483 TI - Separation technique of a mixing of two uncorrelated and perfectly polarized lights with different coherence and polarization properties. AB - The sum of two uncorrelated and totally polarized lights with different coherence and polarization properties usually results in a partially polarized light. It is shown in this paper that the initial totally polarized lights can be recovered from the mixed partially polarized light. The proposed technique is based on coherence analysis and does not require the knowledge of the polarization states or the coherence properties of the initial perfectly polarized beams as long as these properties are different for the two waves. Some practical optical implementations of this technique are discussed on different illustrative applications. PMID- 18382484 TI - Image statistics for surface reflectance perception. AB - Human observers can distinguish the albedo of real-world surfaces even when the surfaces are viewed in isolation, contrary to the Gelb effect. We sought to measure this ability and to understand the cues that might underlie it. We took photographs of complex surfaces such as stucco and asked observers to judge their diffuse reflectance by comparing them to a physical Munsell scale. Their judgments, while imperfect, were highly correlated with the true reflectance. The judgments were also highly correlated with certain image statistics, such as moment and percentile statistics of the luminance and subband histograms. When we digitally manipulated these statistics in an image, human judgments were correspondingly altered. Moreover, linear combinations of such statistics allow a machine vision system (operating within the constrained world of single surfaces) to estimate albedo with an accuracy similar to that of human observers. Taken together, these results indicate that some simple image statistics have a strong influence on the judgment of surface reflectance. PMID- 18382485 TI - Optical analysis of a liquid-crystal switch system based on total internal reflection. AB - A complete methodology using matrix representations for describing light transmission and reflection at an interface between an isotropic medium with high refractive index and a uniaxial birefringent material where total internal reflection (TIR) could happen is described systematically. A new TIR-based liquid crystal (LC) switch system is proposed and investigated in detail by using this analyzing method. The criteria of selection of critical parameters such as LC mixture, waveguide, and operation mode of the LC layer, etc., are discussed. Dependence of transmission on incident angle and dynamic characteristics under an electric field are given for different cell gaps. The results give detailed and useful guidance in the fabrication of the LC switch system. PMID- 18382486 TI - Positive-operator-valued-measure view of the ensemble approach to polarization optics. AB - The statistical ensemble formalism of Kim et al [J. Opt. Soc. Am. A4, 433 (1987)] offers a realistic model for characterizing the effect of stochastic nonimage forming optical media on the state of polarization of transmitted light. With suitable choice of the Jones ensemble, various Mueller transformations-some of which are hitherto unknown-are deduced. It is observed that the ensemble approach is formally identical to the positive-operator-valued measures (POVMs) on the quantum density matrix. This observation, in combination with the recent suggestion by Ahnert and Payne [Phys. Rev. A71, 012330-1 (2005)]-in the context of generalized quantum measurement on single photon polarization states-that linear optics elements can be employed in setting up all possible POVMs enables us to propose a way of realizing different types of Mueller devices. PMID- 18382487 TI - Significant deformations and propagation variations of Laguerre-Gaussian beams reflected and transmitted at a dielectric interface. AB - We observe TM and TE Laguerre-Gaussian (LG) light beams reflected and transmitted at a dielectric interface near critical incidence. The intensity distribution of the reflected beam is transversely deformed near the beam waist, and that of the transmitted beam is similar to that of a diagonal Hermite-Gaussian beam. The former rotates around the optical axis by approximately pi/2 with propagation, and the latter returns to that of the incident LG beam. These observations agree well with numerical calculations based on an angular spectral analysis and are attributable to the helical wavefront of the LG beams, the sharp incidence-angle dependence of the Fresnel reflection and transmission coefficients, and the Gouy phase. PMID- 18382488 TI - Fast method to compute scattering by a buried object under a randomly rough surface: PILE combined with FB-SA. AB - A fast, exact numerical method based on the method of moments (MM) is developed to calculate the scattering from an object below a randomly rough surface. Dechamps et al. [J. Opt. Soc. Am. A23, 359 (2006)] have recently developed the PILE (propagation-inside-layer expansion) method for a stack of two one dimensional rough interfaces separating homogeneous media. From the inversion of the impedance matrix by block (in which two impedance matrices of each interface and two coupling matrices are involved), this method allows one to calculate separately and exactly the multiple-scattering contributions inside the layer in which the inverses of the impedance matrices of each interface are involved. Our purpose here is to apply this method for an object below a rough surface. In addition, to invert a matrix of large size, the forward-backward spectral acceleration (FB-SA) approach of complexity O(N) (N is the number of unknowns on the interface) proposed by Chou and Johnson [Radio Sci.33, 1277 (1998)] is applied. The new method, PILE combined with FB-SA, is tested on perfectly conducting circular and elliptic cylinders located below a dielectric rough interface obeying a Gaussian process with Gaussian and exponential height autocorrelation functions. PMID- 18382489 TI - Refraction and diffraction by a metal-dielectric multilayered structure. AB - An optical refraction prism consisting of metal and dielectric, subwavelength, periodic multilayered thin films has been proposed. The multilayered structure of metal and dielectric thin films has a cylindrical dispersion surface for TM polarized light. The light behaviors are very different from those of conventional glass prisms and photonic crystal superprisms. Refraction and diffraction of the light wave for the metal-dielectric multilayered prism has been investigated by numerical simulations and graphical representation based on the dispersion surface. A prism with 0.2 microm period had an angular dispersion of 0.20 degrees /nm for approximately 0.8 microm wavelength light. The finite thick metal-dielectric multilayered structure acted as a slab waveguide. PMID- 18382490 TI - Image fidelity for single-layer and multi-layer silver superlenses. AB - In response to increasing interest in the area of subdiffraction-limited near field imaging, the performance of several different realizable and theoretical superresolving silver-based lenses is simulated for a variety of different input object profiles. A computationally-efficient T-matrix technique is used to model the lenses, which consist of layers of silver with total width of 40 nm sandwiched between layers of polymethyl methacrylate and silicon dioxide. The lenses are exposed to nonperiodic bright- and dark-slit input patterns, with feature size varied between 1 nm and 2.5 microm. The performance of the lenses is characterized in terms of transfer function, contrast profile, error profile, and input-to-output correlation. It is shown that increasing the number of layers in a lens increases the lens' transmission coefficients at high spatial frequencies; however, this does not always lead to better imaging performance. The main reasons for this are lens-specific resonances that distort features at certain spatial frequencies, and the increased attenuation of the DC component of transmitted images, which reduces image fidelity, particularly for dark-line features. This suggests that, to achieve optimum results, the design of the superresolving lens system should take into account the characteristics of the images that it is expected to transmit. PMID- 18382491 TI - Degree of polarization estimation in the presence of nonuniform illumination and additive Gaussian noise. AB - Within the general framework of active imaging we address the degree of polarization (DOP) estimation in the presence of additive Gaussian detector noise. We first study the performance of standard DOP estimators and propose a method to increase estimation precision using physically relevant a priori information. We then consider the realistic case of nonuniform illumination distribution. We derive the Cramer-Rao lower bound and determine a profile likelihood-based estimator. We demonstrate the efficiency of this new estimator and compare its performance with other standard estimators as a function of the degree of nonuniformity of the illumination. PMID- 18382492 TI - Integral equation formulation for reflection by a mirror. AB - When light is incident on a mirror, it induces a current density on its surface. This surface current density emits radiation, which is the observed reflected field. We consider a monochromatic incident field with an arbitrary spatial dependence, and we derive an integral equation for the Fourier-transformed surface current density. This equation contains the incident electric field at the surface as an inhomogeneous term. The incident field, emitted by a source current density in front of the mirror, is then represented by an angular spectrum, and this leads to a solution of the integral equation. From this result we derive a relation between the surface current density and the current density of the source. It is shown with examples that this approach provides a simple method for obtaining the surface current density. It is also shown that with the solution of the integral equation, an image source can be constructed for any current source, and as illustration we construct the images of electric and magnetic dipoles and the mirror image of an electric quadrupole. By applying the general solution for the surface current density, we derive an expression for the reflected field as an integral over the source current distribution, and this may serve as an alternative to the method of images. PMID- 18382493 TI - Ultrasound-enhanced optical coherence tomography: improved penetration and resolution. AB - Increasing penetration remains one of the most important issues in optical coherence tomography (OCT) research, which we achieved with a parallel ultrasound beam. In addition to qualitative improvements of tissue imaging, quantitative improvements in resolution of up to 28%+/-2% was noted. At lower frequencies and energies the improvement occurred primarily by altering the detection of multiply scattered light (photon-phonon interaction), which was substantially greater in solids than in liquids (even though the liquid had the higher scattering coefficient). In conclusion, the use of an ultrasound beam with OCT appears the most effective means to date for increasing imaging penetration. PMID- 18382494 TI - Resonance Raman imaging of macular pigment distributions in the human retina. AB - We describe resonance Raman imaging (RRI) of macular pigment (MP) distributions in the living human eye. MP consists of the antioxidant carotenoid compounds lutein and zeaxanthin, is typically present in high concentrations in the healthy human macula relative to the peripheral retina, and is thought to protect this important central region from age-related macular degeneration. We demonstrate that RRI is capable of quantifying and imaging the spatially strongly varying MP distribution in the human retina. Using laser excitation of the MP molecules at 488nm, and sequential camera detection of light emitted back from the retina at the MP's strongest Raman peak position and at an off-peak position, RRI maps of MP are obtained at a resolution below 50microm within a fraction of a second per exposure. RRI imaging can be carried out with undilated pupils and provides a highly molecule-specific diagnostic imaging approach for MP distributions in human subjects. PMID- 18382495 TI - Simulations of astronomical imaging phased arrays. AB - We describe a theoretical procedure for analyzing astronomical phased arrays with overlapping beams and apply the procedure to simulate a simple example. We demonstrate the effect of overlapping beams on the number of degrees of freedom of the array and on the ability of the array to recover a source. We show that the best images are obtained using overlapping beams, contrary to common practice, and show how the dynamic range of a phased array directly affects the image quality. PMID- 18382496 TI - Influence of geometrical perturbation at inner boundaries of invisibility cloaks. AB - The influence of a geometrical perturbation delta at the inner boundaries of both cylindrical and spherical invisibility cloaks on invisibility performance is presented. The analytic solutions for such influence in the case of the general coordinate transformation are given. We show that the cylindrical cloak is more sensitive than a spherical cloak to such a perturbation. The difference results from the different asymptotic properties of eigenfunctions for the cylindrical and spherical wave equations. In particular, the zeroth-order scattering coefficient for a cylindrical cloak determined by -1/ln(delta) converges to zero very slowly. The noticeable scattering induced by the slow convergence speed can be decreased by choosing appropriate coordinate transformation functions. More interestingly, the slow convergence can be overcome dramatically by putting a PEC (PMC) layer at the interior boundary of the cloak shell for TM (TE) wave. PMID- 18382497 TI - Eigenfunctions of the complex fractional Fourier transform obtained in the context of quantum optics. AB - We employ the recently established basis (the two-variable Hermite-Gaussian function) of the generalized Bargmann space (BGBS) [Phys. Lett. A303, 311 (2002)] to study the generalized form of the fractional Fourier transform (FRFT). By using the technique of integration within an ordered product of operators and the bipartite entangled-state representations, we derive the generalized generating function of the BGBS with which the undecomposable kernel of the two-dimensional FRFT [also named complex fractional Fourier transform (CFRFT)] is obtained. This approach naturally shows that the BGBS is just the eigenfunction of the CFRFT. PMID- 18382498 TI - Relating vector ray-tracing equations for holograms of arbitrary shape and thickness. AB - The author derives Latta's ray-tracing equations for holograms of arbitrary thickness [Appl. Opt.10, 2698 (1971)] from Welford's vector ray-tracing equation for holograms of arbitrary shape [Opt. Commun.14, 322 (1975)]. The derivation follows Welford's original approach but accounts for changes in the shape and thickness of the recording medium between construction and reconstruction. PMID- 18382499 TI - Phase-error correction in digital holography. AB - The quality of images computed from digital holograms or heterodyne array imaging is degraded by phase errors in the object and/or reference beams at the time of measurement. This paper describes computer simulations used to compare the performance of digital shearing laser interferometry and various sharpness metrics for the correction of such phase errors when imaging a diffuse object. These algorithms are intended for scenarios in which multiple holograms can be recorded with independent object speckle realizations and a static phase error. Algorithm performance is explored as a function of the number of available speckle realizations and signal-to-noise ratio (SNR). The performance of various sharpness metrics is examined in detail and is shown to vary widely. Under ideal conditions with >15 speckle realizations and high SNR, phase corrections better than lambda/50 root-mean-square (RMS) were obtained. Corrections better than lambda/10 RMS were obtained in the high SNR regime with as few as two speckle realizations and at object beam signal levels as low as 2.5 photons/speckle with six speckle realizations. PMID- 18382500 TI - Correction of anisoplanatic phase errors in digital holography. AB - The quality of coherent images computed from digital holography or heterodyne array data is sensitive to phase errors of the reference and/or object beams. A number of algorithms exist for correcting phase errors in or very near the hologram plane. In the case of phase errors introduced a nonnegligible distance away from hologram plane, the resulting imagery exhibits anisoplanatism. A feature of coherent imaging is that such phase errors may be corrected by simply propagating the aberrated fields (from the object) from the hologram plane to the plane where the phase errors were introduced and applying the phase-error correction algorithms to the fields in that plane. We present experimental results that demonstrate correction of such anisoplanatic phase errors. PMID- 18382501 TI - Irreversible effects of random modulation on coherence properties of partially polarized light. AB - The effect of the multiplication of two vector electric fields by random Jones matrices on their coherence properties is investigated by analyzing their interference fringes. It is shown that these random modulations cannot increase the maximal visibility obtained when the polarization states of the two analyzed electric fields are optimized. This new result generalizes a standard property of perfectly polarized light to partially polarized light and is representative of the irreversible effects of random modulations. PMID- 18382502 TI - Efficient and low-noise single-photon detection in 1550 nm communication band by frequency upconversion in periodically poled LiNbO3 waveguides. AB - We demonstrate 1500 nm band single-photon detection with low dark-count noise and a potentially high efficiency, which may allow long distance and high-bit-rate quantum key distribution. By developing frequency upconversion devices based on periodically poled lithium niobate waveguides, which are specifically designed to use a pump wavelength longer than that of communication-band photons, we completely eliminate the dark-count noise caused by parasitic nonlinear processes in the waveguide. We observed an internal conversion efficiency as high as 40% and demonstrated scaling down to the single photon level while maintaining a background dark-count rate of 10(2)s(-1). PMID- 18382503 TI - Can a light beam be considered to be the sum of a completely polarized and a completely unpolarized beam? AB - In a classic paper that may be regarded as the starting point of polarization optics, G. G. Stokes [Trans. Cambridge Philos. Soc.9, 399 (1852)] presented a theorem according to which any light beam is equivalent to the sum of two light beams, one of which is completely polarized and the other completely unpolarized. We show that Stokes' proof of this theorem is flawed. We present a condition for the theorem to be valid. PMID- 18382504 TI - High-contrast, high-intensity laser pulse generation using a nonlinear preamplifier in a Ti:sapphire laser system. AB - We report a high-contrast, high-intensity Ti:sapphire chirped-pulse amplification system that incorporates a nonlinear preamplifier based on optical parametric chirped-pulse amplification (OPCPA). By cooling the Ti:sapphire crystal in the final amplifier down to 77 K, the chirped-pulses are amplified to 2.9 J at a 10 Hz repetition rate without a thermal lensing effect. Pulse compression down to 19 fs duration obtained after amplification indicates a peak power of 80 TW. With the OPCPA, the temporal contrast is significantly improved to better than 7x10( 9) in a few picoseconds interval prior to the main laser pulse. PMID- 18382505 TI - Simulataneous coherent and spectral addition of fiber lasers. AB - A new approach for the simultaneous coherent and spectral addition of a two dimensional array of fiber lasers is presented. A combining efficiency of over 80% and a combined beam profile with M(2)=1.15 were experimentally obtained with an array of four fiber lasers. This approach can lead to significant upscaling in fiber laser additions. PMID- 18382506 TI - Pulse-duration dependency of femtosecond laser refractive index modification in poly(methyl methacrylate). AB - Refractive index modification of pure poly(methyl methacrylate) (PMMA) is investigated as a function of pulse duration using femtosecond lasers at 800 and 387 nm wavelength. It is observed that at 800 nm, the refractive index is modified more efficiently as the pulse duration decreases below 100 fs, whereas at 387 nm, efficient index modification is accomplished with longer, 180 fs pulses. Results suggest that three- and two-photon absorption is responsible for modification of pure PMMA at 800 nm and 387 nm, respectively. Repeated irradiation with short pulses of low laser fluence allows control of the photomodification via incubation, thus reducing bulk damage. PMID- 18382507 TI - Sensor for the detection of local contrast gloss of products. AB - We introduce a sensor for providing information on the local contrast gloss (or luster) of products. The sensor also provides information of the local specular gloss of the object. The signals of this sensor are produced by diffractive optical elements from fields that are scattered in the diffuse and specular directions from the object. We present specular gloss, diffuse-reflectance factor related to the contrast gloss, and visibility maps measured from black prints on paper. High variation can be observed in the relevant gloss parameters obtained from the printed area. In addition, borders of the print can be clearly detected from the diffuse-reflectance-factor maps. The sensor also allows detection of raster points of a print. This sensor opens up entirely new means to study prints and other substances. PMID- 18382508 TI - Electrostatic forward-viewing scanning probe for Doppler optical coherence tomography using a dissipative polymer catheter. AB - A novel flexible scanning optical probe is constructed with a finely etched optical fiber strung through a platinum coil in the lumen of a dissipative polymer. The packaged probe is 2.2 mm in diameter with a rigid length of 6mm when using a ball lens or 12 mm when scanning the fiber proximal to a gradient-index (GRIN) lens. Driven by constant high voltage (1-3 kV) at low current (< 5 microA), the probe oscillates to provide wide forward-viewing angle (13 degrees and 33 degrees with ball and GRIN lens designs, respectively) and high-frame-rate (10-140 fps) operation. Motion of the probe tip is observed with a high-speed camera and compared with theory. Optical coherence tomography (OCT) imaging with the probe is demonstrated with a wavelength-swept source laser. Images of an IR card as well as in vivo Doppler OCT images of a tadpole heart are presented. This optomechanical design offers a simple, inexpensive method to obtain a high-frame rate forward-viewing scanning probe. PMID- 18382509 TI - Low-threshold supercontinuum generation in highly nonlinear chalcogenide nanowires. AB - We demonstrate low-threshold supercontinuum generated in a highly nonlinear arsenic selenide chalcogenide nanowire with tailored dispersion. The tapered submicrometer chalcogenide fiber exhibits an ultrahigh nonlinearity, n(2) approximately 1.1x10(-17) m(2)/W and an effective mode area of 0.48 mum(2), yielding an effective nonlinearity of gamma approximately 93.4 W/m, which is over 80,000 times larger than standard silica single-mode fiber at a wavelength of approximately 1550 nm. This high nonlinearity, in conjunction with the engineered anomalous dispersion, enables low-threshold soliton fission leading to large spectral broadening at a dramatically reduced peak power of several watts, corresponding to picojoule energy. PMID- 18382510 TI - Observation of two-dimensional lattice interface solitons. AB - We report on the experimental observation of two-dimensional solitons at the interface between square and hexagonal waveguide arrays. In addition to the different symmetries of the lattices, the influence of a varying refractive index modulation depth is investigated. Such variation strongly affects the properties of surface solitons residing at different sides of the interface. PMID- 18382511 TI - Four-wave parametric conversion of femtosecond laser pulse in a filament induced in a solid target. AB - Nonlinear interaction of two femtosecond laser pulses in a filament, induced by one of them inside a fused silica plate, leads to generation of the new spectral components. These spectral components reach hundreds of nanometers bandwidth. Their spatial and spectral properties can be explained by four-wave parametric coupling in the filament. The energy measurements indicate the high efficiency of this process. PMID- 18382512 TI - Nd:YAG laser pumped at 946 nm. AB - A Nd:YAG laser crystal was pumped at 946 nm and lased at 1064 nm. This pump-lase format was investigated in order to reduce the quantum defect between the pump and laser photons as compared to other pump schemes of this material. To the best of our knowledge, this is the first realization of this scheme. A room temperature absorption coefficient and linewidth of approximately 0.075 cm(-1) and approximately 1 nm for 1% at. Nd(+3) concentrations were measured for the 946 nm absorption line. Those parameters impose both narrow-bandwidth pumping and a long absorption path. By increasing the laser crystal temperature above room temperature, the absorption cross sections at 946 and 938 nm increase due to enhanced thermal population of the upper energy level of the ground manifold. The possibility of exploiting this phenomenon to enhance the pump absorption is also discussed. PMID- 18382513 TI - Hollow-core waveguide characterization by optically induced particle transport. AB - We introduce a method for optical characterization of hollow-core optical waveguides. Radiation pressure exerted by the waveguide modes on dielectric microspheres is used to analyze salient properties such as propagation loss and waveguide mode profiles. These quantities were measured for quasi-single-mode and multimode propagation in on-chip liquid-filled hollow-core antiresonant reflecting optical waveguides. Excellent agreement with analytical and numerical models is found, demonstrating that optically induced particle transport provides a simple, inexpensive, and nondestructive alternative to other characterization methods. PMID- 18382514 TI - Two-soliton interaction in the vicinity of a defect inside a fiber Bragg grating and its application for obtaining an all-optical memory. AB - We study the interaction between two Bragg solitons in the vicinity of a defect inside a fiber Bragg grating. A soliton that is trapped in the defect can be released by launching a second soliton. The effect can be used to obtain an all optical memory that is not strongly sensitive to the phase and the timing arrival of the solitons. PMID- 18382515 TI - Efficient method for launching in-gap solitons in fiber Bragg gratings using a two-segment apodization profile. AB - We theoretically demonstrate what is a new method for efficient launching of in gap solitons in fiber Bragg gratings. The method is based on generating a soliton outside the grating bandgap. Then, the soliton is adiabatically coupled into the bandgap by using its particlelike behavior. We compare our method to a previously published launching scheme that is based on generating the soliton directly within the grating bandgap. When using low-intensity incident pulses, the transmission efficiency of our method is three times higher than that of the previously published scheme. PMID- 18382516 TI - Tighter focusing with a parabolic mirror. AB - We demonstrate experimentally and theoretically that a parabolic mirror (PM) with a high numerical aperture (NA) of 1 focuses a radially polarized laser mode to the smallest diffraction-limited spot at a fixed NA and wavelength, having an area of 0.134 lambda(2). The measurements were performed with a confocal microscope, using the PM as a focusing and collecting element. The results stand in accordance with the theoretical calculations presented by Davidson and Bokor [Opt. Lett. 29, 1318 (2004)], who predicted a reduction in the total focal spot size of 43% as compared with an aplanatic lens. PMID- 18382517 TI - Geometric analysis of spatial distortion in projection-type integral imaging. AB - In projection-type integral imaging, positional errors in elemental images and elemental lenses affect three-dimensional (3D) image quality. We analyzed the relationships between the geometric distortion in elemental images caused by a projection lens and the spatial distortion in the reconstructed 3D image. As a result, we clarified that 3D images that were reconstructed far from the lens array were largely affected, and that the reconstructed images were significantly distorted in the depth direction at the corners of the displayed images. PMID- 18382518 TI - Investigation of spontaneously generated coherence in dressed states of (85)Rb atoms. AB - We investigate the effects of spontaneously generated coherence (SGC) on the electromagnetically induced transparency features in a four-level Lambda-type system. We show that double-transparency windows and a controllable narrow absorption peak can be obtained with the presence of SGC. We present an equivalent system without the rigorous requirement of close-lying levels to observe the phenomena. We also experimentally demonstrate the corresponding features in a rubidium atomic beam. PMID- 18382519 TI - Ultrafast thulium-doped fiber-oscillator with pulse energy of 4.3 nJ. AB - A femtosecond fiber laser based on thulium-doped double-clad silica fiber with an internal dispersion compensation is presented that generates pulses at a center wavelength of 1976 nm with an energy of 4.3 nJ and a duration of 1.2 ps. The dechirped pulse duration is 294 fs. The pulse energy is more than 2 orders of magnitude above the pulse energy demonstrated previously. Mode locking is achieved using additive pulse mode locking, and dispersion compensation is facilitated by a grating stretcher arrangement. PMID- 18382521 TI - Electro-optically forbidden or enhanced spin-to-orbital angular momentum conversion in a focused light beam. AB - We demonstrate that the conversion of spin-to-orbital angular momentum in a focused vectorial light beam can be forbidden or enhanced by the electric-field erased or increased spatial anisotropy of a uniaxial crystal undergoing the Pockels effect. A focused right-handed circularly polarized quasi-Gaussian beam incident on an electro-optic crystal strontium barium niobate is taken to illustrate the effect, and a conversion-forbidden electric field E(0)=-16.87 kV/cm is found. Of special interest is that single photons generated in this effect exhibit an entanglement of 2 deg of freedom of spin and orbital angular momentum, and it is electro-optically controllable. PMID- 18382520 TI - Enhanced green upconversion emission of Er(3+) through energy transfer by Dy(3+) under 800 nm femtosecond-laser excitation. AB - Er(3+) green upconversion (UC) emission corresponding to the transition of (4)S(3/2) ((2)H(11/2))-->(4)I(15/2) is enhanced in a Er/Dy-codoped LiNbO(3) crystal compared with Er-doped LiNbO(3) under 800 nm femtosecond-laser excitation at room temperature. The upconversion mechanisms are proposed based on spectral, kinetic, and pump-power dependence analyses. The energy-transfer efficiency from Dy(3+)((4)F(9/2)) to Er(3+)((4)F(7/2)) is 33%, which results in the enhancement of green UC emission. This energy transfer is advantageous for the Er(3+) UC emission sensitized by Dy(3+), especially in a low-phonon-energy host matrix. PMID- 18382522 TI - Nonlinear optical response from single spheres coated by a nonlinear monolayer. AB - We detected the second-order nonlinear response from single isolated spheres comprised from a centrosymmetric material but covered by a layer of a material with strong second-order nonlinear properties and isolated from an ensemble by the optical trapping technique. We show that when large size parameter spheres are used, the measured second-harmonic efficiency deviates strongly from the prediction of the nonlinear Rayleigh scattering theory. Our results are in very good agreement with the predictions from the exact nonlinear Mie scattering theory. PMID- 18382523 TI - Continuous wave, 30 W laser-diode bar with 10 GHz linewidth for Rb laser pumping. AB - A laser-diode bar incorporated into an external cavity with a volume Bragg mirror produced 30 W of cw output power within a 20 pm (10 GHz) spectral linewidth (FWHM) centered at 780 nm. The device output power exceeded 90% of that for the free-running laser-diode bar. The emission wavelength was tuned over a 400 pm range without broadening laser spectrum width. Absorption of 90% of the laser radiation by a 25 mm vapor cell containing Rb that has been pressure broadened with 300 torr of ethane was demonstrated. PMID- 18382524 TI - Highly efficient diode-pumped actively Q-switched Nd:YAG-SrWO(4) intracavity Raman laser. AB - A highly efficient diode-pumped actively Q-switched intracavity Raman laser with SrWO(4) as the Raman-active medium is presented. As high as 23.8% diode-to-Stokes optical conversion efficiency is obtained with an incident pump power of 7.17 W and a pulse repetition rate of 15 kHz. PMID- 18382525 TI - Label-free optical biosensing with slot-waveguides. AB - We demonstrate label-free molecule detection by using an integrated biosensor based on a Si(3)N(4)/SiO(2) slot-waveguide microring resonator. Bovine serum albumin (BSA) and anti-BSA molecular binding events on the sensor surface are monitored through the measurement of resonant wavelength shifts with varying biomolecule concentrations. The biosensor exhibited sensitivities of 1.8 and 3.2 nm/(ng/mm(2)) for the detection of anti-BSA and BSA, respectively. The estimated detection limits are 28 and 16 pg/mm(2) for anti-BSA and BSA, respectively, limited by wavelength resolution. PMID- 18382526 TI - Development and preliminary results of an endoscopic Raman probe for potential in vivo diagnosis of lung cancers. AB - A near-infrared Raman system was developed to collect real-time in vivo human lung spectra. The excitation light and the emission were guided to and from the tissue surface by a reusable fiber catheter passed down the instrument channel of a bronchoscope. Two-stage filtering was used to reduce laser noise, fluorescence, and Raman emissions from the fibers. A second fiber bundle guided the emission to a spectrometer where the fibers, in a round packing geometry, were spread out to form a parabolic arc that improved the signal-to-noise ratio 20-fold, facilitating real-time spectral measurements. Preliminary clinical tests show that clear and reliable Raman spectra can be obtained. PMID- 18382527 TI - Integrated Raman- and angular-scattering microscopy. AB - A microscopy system has been constructed that is capable of simultaneously acquiring both traditional Raman spectra as well as angle-resolved elastic scattering patterns using a single focused laser spot less than 10 mum wide. The elastic-scattering signal was analyzed by generalized Lorenz-Mie theory, representing what we believe to be the first experimental validation of the theory's prediction of angular backscatter from single spheres. The microscope system exhibits 3 nm precision in predicting sphere diameters, while simultaneously yielding high-quality Raman signals. Applications to single cell analysis are envisioned. PMID- 18382528 TI - Modal coupling in surface-corrugated long-period-grating fiber tapers. AB - We describe the properties of a surface-corrugated long-period-grating fiber taper fabricated using contact optical lithography and wet etching techniques. The preservation of cylindrical symmetry in this device facilitates investigation of the modal behavior. Comparison of the measured and calculated transmission spectra reveals that the widely used coupled-mode theory is not applicable. Instead, a mode-projection model, in which modal propagation and coupling are treated separately within the grating, explains the experiments very well. PMID- 18382529 TI - Perfect quasi-phase matching for the third-harmonic generation using focused Gaussian beams. AB - We propose a method to realize perfect quasi-phase matching (QPM) for nonlinear optical interactions involving Gaussian beams. Using this method, both the wave vector mismatching and the Gouy phase shift can be compensated. Numerical simulations for the third-harmonic generation show that conversion efficiency near to 100% can be realized even if the fundamental wave is tightly focused, which is difficult or even impossible with the conventional QPM method. PMID- 18382530 TI - Vacuum ultraviolet pulses of 11 fs from fifth-harmonic generation of a Ti:sapphire laser. AB - We demonstrate that in a short Ar cell, generation of the fifth harmonic from 12 fs pulses at 810 nm directly results in ultrashort vacuum UV pulses at 162 nm. They have a spectral width of approximately 5 nm and a duration of 11+/-1 fs (1.4 times the transform limit), as measured by cross correlation with the fundamental pulses. Their energy (estimated to 4 nJ) turned out to be sufficient for use as a pump in time-resolved experiments. PMID- 18382531 TI - Passively mode-locked Yb:YAG thin-disk laser with pulse energies exceeding 13 microJ by use of an active multipass geometry. AB - We demonstrate the generation of high-energy picosecond pulses directly from a thin-disk laser oscillator by employing a self-imaging active multipass geometry. Stable single-pulse operation has been obtained with an average output power in excess of 50 W, excluding a cw background of 8%, at a repetition rate of 3.8 MHz. Self-starting passive mode locking was accomplished using a semiconductor saturable absorber mirror. The maximum pulse energy was 13.4 microJ at a pulse duration of 1.36 ps with a time-bandwidth product of 0.34. Single-pass external frequency doubling with a conversion efficiency of 60% yielded >28 W of average power at 515 nm. PMID- 18382532 TI - Passive mode locking of Yb:KLuW using a single-walled carbon nanotube saturable absorber. AB - Mode locking of an Yb-doped bulk laser in the 1 microm spectral range using a single-walled carbon nanotube saturable absorber (SWCNT-SA) is demonstrated for the first time, to our knowledge. Passive mode locking of an Yb:KLuW laser resulted in nearly transform-limited pulses as short as 115 fs at 1048 nm. In addition, the nonlinear response of the SWCNT-SA was measured, yielding a modulation depth of 0.25% and a relaxation time of 750 fs. PMID- 18382533 TI - Artifact removal in Fourier-domain optical coherence tomography with a piezoelectric fiber stretcher. AB - We describe an artifact removal setup swept-source optical coherence tomography (OCT) system that enables high-speed full-range imaging. We implement a piezoelectric fiber stretcher to generate a periodic phase shift between successive A-scans, thus introducing a transverse modulation. The depth ambiguity is then resolved by performing a Fourier filtering in the transverse direction before processing the data in the axial direction. The dc artifact is also removed. The key factor is that the piezoelectric fiber stretcher can be used to generate discrete phase shifts with a high repetition rate. The proposed experimental setup is a much improved version of the previously reported B-M mode scanning for spectral-domain OCT in that it does not generate additional artifacts. It is a simple and low-cost solution for artifact removal that can easily be applied. PMID- 18382534 TI - Thin-disk Yb:KLu(WO(4))(2) laser with single-pass pumping. AB - Single-pass pumping of a thin disk consisting of an only 50 microm thick epitaxial layer of 32 at.% Yb-doped KLu(WO(4))(2) grown on a 0.35 mm thick undoped KLu(WO(4))(2) substrate is demonstrated. The thin-disk laser delivered 9 W of continuous wave output power near 1030 nm with a slope efficiency of 77%. PMID- 18382535 TI - Simple multiwavelength time-division multiplexed light source for sensing applications. AB - We present a novel multiwavelength, time-division multiplexed laser design that continuously cycles through N spectrally narrow wavelengths, spending a specified, fixed time on each one. The design is based on a matched compressor/stretcher and a custom waveform generator applying modulation preferably to the gain medium. The realization discussed here utilizes a pulsed semiconductor optical amplifier in an all-fiber cavity containing fiber Bragg gratings. The laser cycles through 19 wavelengths in a 44 nm wide spectral band (1333-1377 nm) every 15 micros. The source contains no moving parts, offers high repetition rates, narrow spectral linewidths, and custom spectral profiling of the output. PMID- 18382536 TI - Sub-two-cycle light pulses at 1.6 microm from an optical parametric amplifier. AB - We generate ultrabroadband pulses, spanning the 1200-2100 nm wavelength range, from an 800 nm pumped optical parametric amplifier (OPA) working at degeneracy. We compress the microjoule-level energy pulses to nearly transform-limited 8.5 fs duration by an adaptive system employing a deformable mirror. To our knowledge, these are the shortest light pulses generated at 1.6 microm. PMID- 18382537 TI - Simultaneous cell morphometry and refractive index measurement with dual wavelength digital holographic microscopy and dye-enhanced dispersion of perfusion medium. AB - Digital holographic microscopy (DHM) allows optical-path-difference (OPD) measurements with nanometric accuracy. OPD induced by transparent cells depends on both the refractive index (RI) of cells and their morphology. This Letter presents a dual-wavelength DHM that allows us to separately measure both the RI and the cellular thickness by exploiting an enhanced dispersion of the perfusion medium achieved by the utilization of an extracellular dye. The two wavelengths are chosen in the vicinity of the absorption peak of the dye, where the absorption is accompanied by a significant variation of the RI as a function of the wavelength. PMID- 18382538 TI - Perfect-lens-material condition from adjacent absorptive and gain resonances. AB - We suggest, based on the principle of causality and for a material exhibiting adjacent absorptive and gain resonances, that there can be an intervening frequency where perfect imaging is in theory possible. At this frequency, both the dielectric constant and the permeability are negative, leading to a negative refractive index, and there is no loss. In such a material exhibiting a double resonance, the gain must be at the higher frequency. Through appropriate tuning of the refractive index, all propagating and evanescent fields from the object could then in principle be reconstructed at the image plane, subject to practical implementation limits. PMID- 18382539 TI - Sub-2 fs pulses generated by self-channeling in the deep ultraviolet. AB - The generation of sub-2 fs light pulses in the UV is numerically demonstrated, using frequency conversion in filamentation regime. Few-cycle pulses emitted at 266 nm keep their temporal shape over several tens of centimeters. Self compression results from the interplay between Kerr self-focusing and a low density plasma, which continuously defocuses the pulse over extended propagation ranges. PMID- 18382540 TI - Structured lens formed by a 2D square hole array in a metallic film. AB - A method is proposed to modulate phase using variant square holes in a metallic film based on a fundamental mode approximation model. Phase retardation through square holes in a subwavelength scale in a thin metal film has been analyzed and calculated. Based on the model, a structured lens with a numerical aperture of 0.583 and a focal length of 240 microm formed by a 2D square hole array in a metallic film is designed. Numerical simulation using the finite-difference time domain method is carried out, and the results agree with the theoretical analysis. A focal spot close to the diffraction limit can be obtained. PMID- 18382542 TI - Extension of depth of field using amplitude and phase modulation of the pupil function. AB - We analyze the extension of depth of field using both amplitude and phase modulation of the pupil function. In particular, we discuss the advantages and disadvantages of each approach and establish the range of applicability of each method based on the range of spatial frequencies of interest in the imaging system. To the best of our knowledge, this is the first such report on the range of applicability of amplitude and phase modulation to extend the depth of field. PMID- 18382544 TI - Thin infrared imaging systems through multichannel sampling. AB - The size of infrared camera systems can be reduced by collecting low-resolution images in parallel with multiple narrow-aperture lenses rather than collecting a single high-resolution image with one wide-aperture lens. We describe an infrared imaging system that uses a three-by-three lenslet array with an optical system length of 2.3 mm and achieves Rayleigh criteria resolution comparable with a conventional single-lens system with an optical system length of 26 mm. The high resolution final image generated by this system is reconstructed from the low resolution images gathered by each lenslet. This is accomplished using superresolution reconstruction algorithms based on linear and nonlinear interpolation algorithms. Two implementations of the ultrathin camera are demonstrated and their performances are compared with that of a conventional infrared camera. PMID- 18382545 TI - Signal-processing approaches for image-resolution restoration for TOMBO imagery. AB - Thin observation module by bounded optics (TOMBO) is an optical system that achieves compactness and thinness by replacing a conventional large full aperture by a lenslet array with several smaller apertures. This array allows us to collect diverse low-resolution measurements. Finding an efficient way of combining these diverse measurements to make a high-resolution image is an important research problem. We focus on finding a computational method for performing the resolution restoration and evaluating the method via simulations. Our approach is based on advanced signal-processing concepts: we construct a computational data model based on Fourier optics and propose restoration algorithms based on minimization of an information-theoretic measure, called Csiszar's I divergence between two nonnegative quantities: the measured data and the hypothetical images that are induced by our algorithms through the use of our computational data model. We also incorporate Poisson and Gaussian noise processes to model the physical measurements. To solve the optimization problem, we adapt the popular expectation-maximization method. These iterative algorithms, in a multiplicative form, preserve powerful nonnegativity constraints. We further incorporate a regularization based on minimization of total variation to suppress incurring artifacts such as roughness on the surfaces of the estimates. Two sets of simulation examples show that the algorithms can produce very high-quality estimates from noiseless measurements and reasonably good estimates from noisy measurements, even when the measurements are incomplete. Several interesting and useful avenues for future work such as the effects of measurement selection are suggested in our conclusional remarks. PMID- 18382546 TI - Joint digital-optical design of superresolution multiframe imaging systems. AB - Typical electro-optic imaging systems produce image aliasing artifacts. Superresolution algorithms process multiple aliased images to yield a single high resolution image. We design imaging systems by jointly optimizing the optics and postprocessing to maximize such multiframe imaging performance. We describe efficient software methods that can be used to perform joint design by use of commercially available lens design software. PMID- 18382547 TI - Adaptive framework for robust high-resolution image reconstruction in multiplexed computational imaging architectures. AB - In multiplexed computational imaging schemes, high-resolution images are reconstructed by fusing the information in multiple low-resolution images detected by a two-dimensional array of low-resolution image sensors. The reconstruction procedure assumes a mathematical model for the imaging process that could have generated the low-resolution observations from an unknown high resolution image. In practical settings, the parameters of the mathematical imaging model are known only approximately and are typically estimated before the reconstruction procedure takes place. Violations to the assumed model, such as inaccurate knowledge of the field of view of the imagers, erroneous estimation of the model parameters, and/or accidental scene or environmental changes can be detrimental to the reconstruction quality, even if they are small in number. We present an adaptive algorithm for robust reconstruction of high-resolution images in multiplexed computational imaging architectures. Using robust M-estimators and incorporating a similarity measure, the proposed scheme adopts an adaptive estimation strategy that effectively deals with violations to the assumed imaging model. Comparisons with nonadaptive reconstruction techniques demonstrate the superior performance of the proposed algorithm in terms of reconstruction quality and robustness. PMID- 18382548 TI - Adaptive feature-specific imaging: a face recognition example. AB - We present an adaptive feature-specific imaging (AFSI) system and consider its application to a face recognition task. The proposed system makes use of previous measurements to adapt the projection basis at each step. Using sequential hypothesis testing, we compare AFSI with static-FSI (SFSI) and static or adaptive conventional imaging in terms of the number of measurements required to achieve a specified probability of misclassification (Pe). The AFSI system exhibits significant improvement compared to SFSI and conventional imaging at low signal to-noise ratio (SNR). It is shown that for M=4 hypotheses and desired Pe=10(-2), AFSI requires 100 times fewer measurements than the adaptive conventional imager at SNR= -20 dB. We also show a trade-off, in terms of average detection time, between measurement SNR and adaptation advantage, resulting in an optimal value of integration time (equivalent to SNR) per measurement. PMID- 18382549 TI - Robust, common path, phase shifting interferometer and optical profilometer. AB - We describe an improved implementation of our previously reported common-path, phase shifting, and shearing interferometer. Using a time-multiplexed phase shifting scheme, we demonstrate higher sampling resolution, better light sensitivity, and use of arbitrary phase shifting algorithms. We describe microscopic imaging of the surface profile of a copper-plated silicon wafer and demonstrate that the system is vibration insensitive with approximately lambda/100 repeatability. In a more general discussion of our method, we describe the different functional elements and suggest alternative designs and improvements. Possible uses include full-field coherent imaging and high dynamic range wavefront sensing, which we briefly discuss. PMID- 18382550 TI - Single disperser design for coded aperture snapshot spectral imaging. AB - We present a single disperser spectral imager that exploits recent theoretical work in the area of compressed sensing to achieve snapshot spectral imaging. An experimental prototype is used to capture the spatiospectral information of a scene that consists of two balls illuminated by different light sources. An iterative algorithm is used to reconstruct the data cube. The average spectral resolution is 3.6 nm per spectral channel. The accuracy of the instrument is demonstrated by comparison of the spectra acquired with the proposed system with the spectra acquired by a nonimaging reference spectrometer. PMID- 18382551 TI - Image identification system based on an optical broadcast neural network and a pulse coupled neural network preprocessor stage. AB - We describe the concept of a vision system based on an optoelectronic hardware neural processor. The proposed system is composed of a pulse coupled neural network (PCNN) preprocessor stage that converts an input image into a temporal pulsed pattern. These pulses are inputs to the optical broadcast neural network (OBNN) processor, which classifies the input pattern between a set of reference patterns based on a pattern matching strategy. The PCNN is to provide immunity to the scale, rotation, and translation of objects in the image. The OBNN provides high parallelism and a high speed hardware neural processor. PMID- 18382552 TI - Theoretical foundations for joint digital-optical analysis of electro-optical imaging systems. AB - We describe the mathematical and conceptual foundations for a novel methodology for jointly optimizing the design and analysis of the optics, detector, and digital image processing for imaging systems. Our methodology is based on the end to-end merit function of predicted average pixel sum-squared error to find the optical and image processing parameters that minimize this merit function. Our approach offers several advantages over the traditional principles of optical design, such as improved imaging performance, expanded operating capabilities, and improved as-built performance. PMID- 18382553 TI - Multichannel sampling schemes for optical imaging systems. AB - We introduce a framework of focal-plane coding schemes for multichannel sampling in optical systems. A particular objective is to develop an ultrathin imager without compromising image resolution. We present a complete f/2.1 optical system with a thickness of 2.2 mm. The resolution is maintained in the thin optical system by an integrated design of the encoding scheme, the process of making the coding elements, and the decoding algorithms. PMID- 18382554 TI - Computational imaging systems: joint design and end-to-end optimality. AB - A framework is proposed for optimal joint design of the optical and reconstruction filters in a computational imaging system. First, a technique for the design of a physically unconstrained system is proposed whose performance serves as a universal bound on any realistic computational imaging system. Increasing levels of constraints are then imposed to emulate a physically realizable optical filter. The proposed design employs a generalized Benders' decomposition method to yield multiple globally optimal solutions to the nonconvex optimization problem. Structured, closed-form solutions for the design of observation and reconstruction filters, in terms of the system input and noise autocorrelation matrices, are presented. Numerical comparison with a state-of-the art optical system shows the advantage of joint optimization and concurrent design. PMID- 18382555 TI - Computational optical sensing and imaging: introduction to the feature issue. PMID- 18382556 TI - Photonic-crystal-based cloaking device at optical wavelengths. AB - We present a photonic crystal cloaking device at optical wavelengths based on the association of two lattices working in different regimes, namely, stop band and negative refraction. The idea is to reconstruct in phase an incident cut Gaussian modulated plane wave by using the photonic crystal dispersion properties to ensure that no light penetrates in the core of the device. It is believed that such a cloaking device could become a building block for future generations of 3D integrated optical circuits. PMID- 18382557 TI - Characterization of an active control system for holographic setup stabilization. AB - We report on the realization and characterization of an active control system of the optical holographic setup used for fabrication of holographic gratings in liquid-crystalline composite materials. The system exploits a reference diffraction grating and a piezomirror in closed-loop feedback. The piezoelectric mirror exhibits a hysteresis that depends not only on the applied voltage, but also on the history of the mirror motion. In an open-loop configuration, the hysteresis can be reduced by adjusting the delay time between the application of two different control voltage values; in a closed-loop operation, it is possible to eliminate the residual hysteresis. By testing the system in different conditions, it has been shown that residual fluctuations are comparable to the resolution of the piezomirror operation. PMID- 18382558 TI - Transferring the calibration of direct solar irradiance to diffuse-sky radiance measurements for CIMEL Sun-sky radiometers. AB - Two types of sunphotometric measurement are considered in this study: direct-Sun irradiance and diffuse-sky radiance. Based on CIMEL CE318 Sun-sky radiometer characteristics, we introduce a gain-corrected solid angle that allows interconverting calibration coefficients of these two types of measurement, thus realizing a "vicarious" radiance calibration. The accuracy of the gain-corrected solid angle depends on the number of available historical calibration records. The method is easy to use, provided that at least one laboratory calibration has been made previously. Examples coming from three distinct CE318 versions belonging to the AERONET/PHOTONS network are presented to provide details on the vicarious calibration method and protocols. From the error propagation analysis and the comparison with laboratory results, the uncertainty of the vicarious radiance calibration is shown to be comparable with the laboratory one, e.g., 3% 5%. PMID- 18382559 TI - Adaptive helical mirror for generation of optical phase singularity. AB - We report a specially designed adaptive mirror that can be bent into a helical shape for generation of an optical phase singularity. The adaptive helical mirror (AHM) reported here is a reflective device that can provide a continuous phase variation of the optical field in the azimuthal direction. The construction details and evaluation of the AHM are presented. A Michelson interferometer is used for the detection of the phase singularity. The AHM can be used for generation of a singular beam having multiple topological charges, positive or negative, just by controlling the excitation voltage of the AHM. PMID- 18382561 TI - Comment on "National Ignition Facility laser performance status". AB - The National Ignition Facility (NIF) laser has not yet achieved two of the stated requirements needed for testing and evaluating ignition targets. The laser focal spot size is more than a factor of 2 too large, and the laser bandwidth is more than a factor of 2 too small. PMID- 18382562 TI - Infrared, spectral, directional-hemispherical reflectance of fused silica, Teflon polytetrafluoroethylene polymer, chrome oxide ceramic particle surface, Pyromark 2500 paint, Krylon 1602 paint, and Duraflect coating. AB - Infrared, spectral, directional-hemispherical reflectivity measurements of polished fused silica, Teflon polytetrafluoroethylene polymer, chrome oxide ceramic particle surface, Pyromark 2500 paint, Krylon 1602 paint, and Duraflect coating are provided. The reflectance was measured with an estimated accuracy of 0.01 to 0.02 units and a precision of 0.005 units. All the surfaces were measured at ambient temperatures. Additionally, the chrome oxide ceramic particle surface was measured at 486 K and the Pyromark 2500 at four temperatures to 877 K. Polarization measurements are also provided for fused silica, Duraflect, chrome oxide ceramic particle surface, and Pyromark 2500 paint. Separate diffuse and specular reflectance components for the Duraflect and chrome oxide ceramic surfaces are included. Fresnel-based predictions for fused silica parallel and perpendicular polarized reflections are compared to measurements. It is notable that the Pyromark 2500 and chrome oxide ceramic particle surfaces exhibit a significant lack of manufacturing repeatability. PMID- 18382563 TI - Efficient generation of >2 W of green light by single-pass frequency doubling in PPMgLN. AB - We report 32% efficient frequency doubling of single-frequency 1029 nm light to green light at 514.5 nm using a single-pass configuration. A congruent composition, periodically poled magnesium-doped lithium niobate (PPMgLN) crystal of 50 mm length was used to generate a second-harmonic power of 2.3 W. To our knowledge, this is the highest reported power and efficiency achieved in the second-harmonic generation of single-frequency green light in a single-pass configuration. PMID- 18382564 TI - Active optics: single actuator principle and angular thickness distribution for astigmatism compensation by elasticity. AB - Active optics techniques providing high quality large astigmatism compensations to be used in a fixed or variable optical path, are presented in three ways: analytical calculations, finite element analysis, and experimental validation. The use of an angular thickness distribution allows improving the optical quality of toroidal deformations. We emphasize the case of a single actuator design, in which the principle is proven with a prototype. PMID- 18382565 TI - Numerical and experimental study of microfluidic devices in step-index optical fibers. AB - Microfluidic devices composed of microslits in step-index optical fibers are thoroughly investigated. Numerical simulations are performed to explain scattering and power loss in such devices. Experimental results based on microslits fabricated by femtosecond laser processing corroborate theoretical data. Dependency of the device performance on the refractive index of fluid in the slit is further utilized to construct a refractive index sensor and an in fiber attenuator. PMID- 18382566 TI - Subsurface damage distribution in the lapping process. AB - To systematically investigate the influence of lapping parameters on subsurface damage (SSD) depth and characterize the damage feature comprehensively, maximum depth and distribution of SSD generated in the optical lapping process were measured with the magnetorheological finishing wedge technique. Then, an interaction of adjacent indentations was applied to interpret the generation of maximum depth of SSD. Eventually, the lapping procedure based on the influence of lapping parameters on the material removal rate and SSD depth was proposed to improve the lapping efficiency. PMID- 18382567 TI - High-fidelity replication of Dammann gratings using soft lithography. AB - We report the experimental results of using the soft lithography method for replication of Dammann gratings. By using an elastomeric stamp, uniform grating structures were transferred to the UV-curable polymer. To evaluate the quality of the replication, diffraction images and light intensity were measured. Compared with the master devices, the replicas of Dammann gratings show a slight deviation in both surface relief profile and optical performance. Experimental results demonstrated that high-fidelity replication of Dammann gratings is realized by using soft lithography with low cost and high throughput. PMID- 18382568 TI - Design and fabrication of diffractive microlens arrays with continuous relief for parallel laser direct writing. AB - Diffractive microlens arrays with continuous relief are designed, fabricated, and characterized by using Fermat's principle to create an array of spots on the photoresist-coated surface of a substrate for parallel laser direct writing. Experimental results indicate that a diffraction efficiency of 71.4% and a spot size of 1.97 microm (FWHM) can be achieved at normal incidence and a writing laser wavelength of 441.6 nm with an array of F/4 fabricated on fused silica, and the developed array can be used to improve the utilization ratio of writing laser energy. PMID- 18382569 TI - Coherent light scattering on nanofluids: computer simulation results. AB - If coherent light is incident on a suspension containing nanoparticles, they act as scattering centers and the result of the far-field interference is a "speckled" image. The scattering centers have a complex movement of both sedimentation and Brownian motion. Consequently the speckle image is not static but presents time fluctuations. A computer code to simulate the dynamics of the coherent light scattering on nanofluids was written, tested, and used to calculate the far-field intensity variation for nanofluids having different particle size. The results are discussed and an alternative experimental method for fast nanoparticle size assessing is suggested as a possible application. PMID- 18382570 TI - Hyper-Rayleigh scattering with picosecond pulse trains. AB - We propose a method for measuring hyper-Rayleigh scattering employing pulse trains produced by a Q-switched and mode-locked Nd:YAG laser. The use of the entire pulse train under the Q-switch envelope avoids the need of any device to scan the irradiance, as is usually done with nanosecond and femtosecond single pulse lasers. To verify the feasibility of the technique, we performed measurements in different solutions of para-nitroaniline and compared the results with those obtained with nanosecond pulses. In both cases, the agreement with the hyperpolarizability values reported in the literature is about the same, but the measurements carried out with pulse trains are at least 20 times faster. Besides the advantage of acquisition speed, the use of pulse trains also allows the instantaneous inspection of slow luminescence contributions arising from multiphoton absorption. PMID- 18382571 TI - Tunable 6.8 W narrow bandwidth emission from a single-stripe continuous-wave broad-area laser diode in a simple external cavity. AB - An antireflection-coated broad-area laser diode with an emitter size of 400 microm x 1 microm and a chip length of 1500 microm is operated in a simple external cavity. For wavelength stabilization and to narrow the bandwidth a diffraction grating in a Littrow configuration is used. At an injection current of 9 A up to 6.8 W of optical output power and a resulting slope efficiency of 0.8 W/A could be achieved. Further, the bandwidth could be narrowed to 100 pm (FWHM), and a tuning range of 40 nm around 976 nm was obtained. PMID- 18382572 TI - GaN membrane metal-semiconductor-metal ultraviolet photodetector. AB - GaN is a wide-bandgap semiconductor with still unexplored capabilities for ultraviolet detection. To exploit GaN properties better for ultraviolet detection, a metal-semiconductor-metal-type photodetector structure was designed and manufactured on a 2.2 microm thin GaN membrane fabricated by micromachining techniques. As a result, a very low dark current (30 pA at 3 V) and a maximum responsivity of 14 mA/W at a wavelength of 370 nm were obtained. PMID- 18382573 TI - Simultaneous measurement of refractive index and thickness of birefringent wave plates. AB - A nondestructive measurement system based on a position sensing detector (PSD) and a laser interferometer for determining the thickness and refractive indices of birefringent optical wave plates has been developed. Unlike previous methods presented in the literature, the proposed metrology system allows the refractive index and thickness properties of the optical plate to be measured simultaneously. The experimental results obtained for the e-light and o-light refractive indices of a commercially available birefringent optical wave plate with refractive indices of n(o)=1.542972 and n(e)=1.552033 are found to be accurate to within 0.004132 and 0.000229, respectively. Furthermore, the experimentally derived value of the wave plate thickness deviates by no more than 0.9 microm from the analytically derived value of 453.95 microm. Overall, the experimental results confirm that the proposed metrology system provides a simple yet highly accurate means of obtaining simultaneous measurements of the refractive indices and thickness of birefringent optical wave plates. PMID- 18382574 TI - Continuous phase plate for laser beam smoothing. AB - We discuss the beam smoothing principle of a continuous phase plate (CPP) while the input light is varying. The analysis model of the process in which the laser beam with random phase noise propagates through a CPP has been established. With this model the beam smoothing mechanism of the CPP for the laser beam with a different phase aberrations can be described. A method to optimize the smoothing result is introduced. PMID- 18382575 TI - Invariant grating pseudoimaging using polychromatic light and a finite extension source. AB - The Talbot effect is a well studied phenomenon by which grating pseudoimages appear at certain periodic distances when monochromatic light is used. Recently, numerical simulations have shown a new phenomenon; when a polychromatic light beam is used in a double grating system, the intensity of the pseudoimages presents a transverse-profile that remains unaffected over a wide range of propagation distances. This effect can be used to increase the tolerances of gratings based optical devices, such as displacement measurement systems, interferometers, and spectrometers. The pseudoimages formation with a polychromatic and finite extension light source is analytically and experimentally demonstrated. Relatively simple analytical expressions for the intensity and the contrast allow us to predict when pseudoimages present a constant contrast and when they disappear. Furthermore, we experimentally obtain the pseudoimages using the proposed configuration, corroborating the theoretical predictions. PMID- 18382576 TI - Measurements of cirrus cloud backscatter color ratio with a two-wavelength lidar. AB - We present observations of cirrus clouds from June 2006 to July 2007 performed by using a two-wavelength lidar located at Hampton University. For this time period, cirrus clouds were observed mostly in 7-13.5 km altitudes. Data analyses have been performed focusing on a color-ratio retrieval. In total, 86,369 samples from 1,689 profiles (1 min average and 15 m range resolution) containing cirrus clouds with attenuated backscatter ratio (ratio of attenuated total backscatter to the molecular backscatter) larger than 10 have been selected. The cirrus color ratio distribution shows a peak value at about 0.88 and a full width at half-maximum of 0.12. PMID- 18382577 TI - Heterodyne detection with a weak local oscillator. AB - Heterodyne detection in the limit of weak (a few photons) local oscillator and signal power levels has been largely neglected in the past, as authors almost always assumed that the noise was dominated by the shot noise from a strong local oscillator. We present the theory for heterodyne detection of diffuse and specular targets at arbitrary power levels, including the case where the local oscillator power is only a few photons per coherent integration period. The theory was tested with experimental results, and was found to show good agreement. We show how to interpret the power spectral density of the heterodyne signal and how to determine the optimal number of signal and local oscillator photons per coherent integration. PMID- 18382578 TI - Automated measurement of the refractive index of fluids. AB - A method for automating refractive-index measurements of fluids has been developed. An encoded rotation stage and position-sensitive detector enable automated reading of angles typically acquired by visual means. Two tunable lasers are used to obtain index measurements at ten discrete wavelengths across the visible spectrum. This method has been implemented on a Hilger-Chance refractometer from which the bulk refractive-index values for various transparent fluids have been measured. An index measurement accuracy of better than one part in the fourth decimal place for distilled water and a few parts in the fourth decimal place for higher index fluids is obtained. PMID- 18382579 TI - Lensed fiber probes designed as an alternative to bulk probes in optical coherence tomography. AB - We demonstrate a compact all-fiber sampling probe for an optical coherence tomography (OCT) system. By forming a focusing lens directly on the tip of an optical fiber, a compact sampling probe could be implemented. To simultaneously achieve a sufficiently long working distance and a good lateral resolution, we employed a large-mode area photonic crystal fiber (PCF) and a coreless silica fiber (CSF) of the same diameters. A working distance of up to 1270 microm, a 3 dB distance range of 2210 microm, and a transverse resolution of 14.2 microm were achieved with the implemented PCF lensed fiber; these values are comparable to those obtainable with a conventional objective lens having an NA of 0.25 (10 x). The performance of the OCT system equipped with the proposed PCF lensed fiber is presented by showing the OCT images of a rat finger as a biological sample and a pearl as an in-depth sample. PMID- 18382580 TI - Self-seeding of a pulsed double-grating Ti:sapphire laser oscillator. AB - A self-seeded pulsed double-grating Ti:sapphire laser oscillator consisting of a grazing incidence cavity geometry with a pair of gratings and a standing-wave cavity pumped by a frequency-doubled Nd:YAG laser was developed and characterized. With self-seeding, narrow-linewidth single-longitudinal-mode (SLM) operation and SLM scanning were possible with a reduced lasing threshold, which was desirable for the intended applications. PMID- 18382581 TI - Development of an eye-safe solid-state tunable laser transmitter in the 1.4-1.5 microm wavelength region based on Cr4+:YAG crystal for lidar applications. AB - An experimental optimization of the efficiency of a gain switched tunable Cr4+:YAG laser at 10 Hz is described. The thermal lensing during pulsed operation was measured. Optimal performance occurred at a crystal temperature of 34 degrees C and resulted in an output energy of approximately 7 mJ and a pulse duration of approximately 35 ns. Tunability in the range of 1350-1500 nm, spectral linewidth of approximately 200 GHz, and M2<4 are demonstrated. The main laser material parameters are estimated. Such a laser could be employed in a laboratory-based nonscanning lidar system if a narrowband birefringent filter is installed. The tunability will permit the improvement of the Cr4+:YAG transmitter for water vapor differential absorption lidar if injection seeding is applied. PMID- 18382582 TI - Projection display for the generation of two orthogonal polarized images using liquid crystal on silicon panels and light emitting diodes. AB - We present a projection system that is capable of two-dimensional and three dimensional image display. A novel projection architecture is discussed that can simultaneously generate two linear polarized full-color images with orthogonal states of polarization using only one optical system. Both images are modulated by using two high-resolution liquid crystal on silicon panels that are illuminated with high-power light emitting diodes. The optical core and the illumination system are simulated, characterized, and optimized with nonsequential ray tracing software. A proof-of-concept demonstrator of the entire projection system is built and characterized. Important component specifications are discussed to improve the system performance. PMID- 18382583 TI - Sub-parts-per-billion level detection of dimethyl methyl phosphonate (DMMP) by quantum cascade laser photoacoustic spectroscopy. AB - The need for the detection of chemical warfare agents (CWAs) is no longer confined to battlefield environments because of at least one confirmed terrorist attack, the Tokyo Subway [Emerg. Infect. Dis. 5, 513 (1999)] in 1995, and a suspected, i.e., a false-alarm of a CWA in the Russell Senate Office Building [Washington Post, 9 February 2006, p. B01]. Therefore, detection of CWAs with high sensitivity and low false-alarm rates is considered an important priority for ensuring public safety. We report a minimum detection level for a CWA simulant, dimethyl methyl phosphonate (DMMP), of <0.5 ppb (parts in 10(9)) by use of a widely tunable external grating cavity quantum cascade laser and photoacoustic spectroscopy. With interferents present in Santa Monica, California street air, we demonstrate a false-alarm rate of 1:10(6) at a detection threshold of 1.6 ppb. PMID- 18382584 TI - Long-period grating fabricated by periodically tapering standard single-mode fiber. AB - We fabricated an asymmetric long-period grating (LPG) by periodically tapering a section of standard single-mode fiber using a resistive filament heating. The LPG exhibits large peak transmission attenuation of -30.31 dB with only 22 periods in a 1.0 cm long optical fiber and possesses unique characteristics for sensing applications. The bending and strain sensitivities are 1.74 nm m and 1.11 pm/mu epsilon, respectively. The polarization dependent loss is large, up to 11.65 dB, which is caused by an asymmetric index profile in the cross section of the tapered LPG. PMID- 18382585 TI - Diffractive optical elements for simultaneous operation in reflection and transmission. AB - It is advantageous for some diffractive optical element (DOE) applications to produce different output patterns in different circumstances. There has been considerable work on the design of wavelength multiplexing DOEs and in devices where the polarization of the incident light determines the output. One parameter that has not, to our knowledge, been exploited for pattern formation DOEs is the mode of operation, i.e., whether the element works in reflection or transmission. We present an approach for designing such devices and design an element with modeled efficiency, mean square error (MSE), and cross-talk of 65.9, 2.52, and 4.2% in transmission and 66.6, 2.50, and 3.5% in reflection. The element has been successfully fabricated and has measured efficiencies of 58.3% +/- 2 in reflection and 68.8% +/- 5 in transmission are reported. PMID- 18382586 TI - Detector effects in photothermal deflection experiments. AB - We report on the theoretical analysis of a detector type influence on the normal deflection signal in photothermal experiments. Two cases are examined. In the first, the quadrant photodiode was considered as the detector; in the second the signal from the position detector, which measures the central moment displacement of the probe beam, was analyzed. Both analyses were carried out within the framework of the complex ray theory. The normal photodeflection signal was found to depend on the type of detector used in the photothermal deflection experiments for some parameters of its setup. PMID- 18382587 TI - Computer generated holograms from three dimensional meshes using an analytic light transport model. AB - We present a method to analytically compute the light distribution of triangles directly in frequency space. This allows for fast evaluation, shading, and propagation of light from 3D mesh objects using angular spectrum methods. The algorithm complexity is only dependent on the hologram resolution and the polygon count of the 3D model. In contrast to other polygon based computer generated holography methods we do not need to perform a Fourier transform per surface. The theory behind the approach is derived, and a suitable algorithm to compute a digital hologram from a general triangle mesh is presented. We review some first results rendered on a spatial-light-modulator-based display by our proof-of concept software. PMID- 18382588 TI - Q(M) and the depolarization index scalar metrics. AB - A depolarization scalar metric for Mueller matrices, named Q(M), is derived from the degree of polarization. Q(M) has been recently reported, and it has been deduced from the nine bilinear constraints between the sixteen elements of the Mueller-Jones matrix. We discuss the relations between Q(M) and the depolarization index. PMID- 18382589 TI - Broadband emission from a multicore fiber fabricated with granulated oxides. AB - We demonstrate a multicore multidopant fiber which, when pumped with a single pump source around approximately 800 nm, emits a more than one octave-spanning fluorescence spectrum ranging from 925 to 2300 nm. The fiber preform is manufactured from granulated oxides and the individual cores are doped with five different rare earths, i.e., Nd3+, Yb3+, Er3+, Ho3+, and Tm3+. PMID- 18382590 TI - Metameric interference security image structures. AB - We study innovative interference security image structures based on metamerism. We have designed, fabricated, and evaluated different structures that can be used in transmission or in reflection. These metameric structures are either a combination of two different interference filters or of an interference filter and a noniridescent colored material. In the latter case, by closely matching the spectra, the sensitivity of the device to changes in light sources and observers is minimized. Because of the intrinsic color shift of interference filters, one can create a hidden image that appears at a specific observation angle. The presence of the hidden image, as well as in some cases of the noniridescent material, which serves as a color reference, increases the complexity of such devices while facilitating the user's authentication process as well as automatized detection by using a laser at a specific angle. We present the design approach, analyze the filters' sensitivity to deposition errors, and evaluate the performance of prototype devices prepared by dual ion beam sputtering. PMID- 18382591 TI - Charge-coupled device spectrograph for direct solar irradiance and sky radiance measurements. AB - The characterization of a charged-coupled device (CCD) spectrograph developed at the Laboratory of Atmospheric Physics, Thessaloniki is presented. The absolute sensitivity of the instrument for direct irradiance and sky radiance measurements was determined, respectively, with an uncertainty of 4.4% and 6.6% in the UV-B, and 3% and 6% in the UV-A, visible and near-infrared (NIR) wavelength ranges. The overall uncertainty associated with the direct irradiance and the sky radiance measurements is, respectively, of the order of 5% and 7% in the UV-B, increasing to 10% for low signals [e.g., at solar zenith angles (SZAs) larger than 70 degrees ], and 4% and 6% in the UV-A, visible, and NIR. Direct solar spectral irradiance measurements from an independently calibrated spectroradiometer (Bentham DTM 300) were compared with the corresponding CCD measurements. Their agreement in the wavelength range of 310-500nm is within 0.5% +/- 1.1% (for SZA between 20 degrees and 70 degrees ). Aerosol optical depth (AOD) derived by the two instruments using direct Sun spectra and by a collocated Cimel sunphotometer [Aerosol Robotic network (AERONET)] agree to within 0.02 +/- 0.02 in the range of 315-870 nm. Significant correlation coefficients with a maximum of 0.99 in the range of 340-360 nm and a minimum of 0.90 at 870 nm were found between synchronous AOD measurements with the Bentham and the Cimel instruments. PMID- 18382592 TI - Multispectral fluorescence lifetime imaging of feces-contaminated apples by time resolved laser-induced fluorescence imaging system with tunable excitation wavelengths. AB - We recently developed a time-resolved multispectral laser-induced fluorescence (LIF) imaging system capable of tunable wavelengths in the visible region for sample excitation and nanosecond-scale characterizations of fluorescence responses (lifetime imaging). Time-dependent fluorescence decay characteristics and fluorescence lifetime imaging of apples artificially contaminated with a range of diluted cow feces were investigated at 670 and 685 nm emission bands obtained by 418, 530, and 630 nm excitations. The results demonstrated that a 670 nm emission with a 418 nm excitation provided the greatest difference in time dependent fluorescence responses between the apples and feces-treated spots. The versatilities of the time-resolved LIF imaging system, including fluorescence lifetime imaging of a relatively large biological object in a multispectral excitation-emission wavelength domain, were demonstrated. PMID- 18382593 TI - Bayesian assessment of uncertainty in aerosol size distributions and index of refraction retrieved from multiwavelength lidar measurements. AB - We investigate the assessment of uncertainty in the inference of aerosol size distributions from backscatter and extinction measurements that can be obtained from a modern elastic/Raman lidar system with a Nd:YAG laser transmitter. To calculate the uncertainty, an analytic formula for the correlated probability density function (PDF) describing the error for an optical coefficient ratio is derived based on a normally distributed fractional error in the optical coefficients. Assuming a monomodal lognormal particle size distribution of spherical, homogeneous particles with a known index of refraction, we compare the assessment of uncertainty using a more conventional forward Monte Carlo method with that obtained from a Bayesian posterior PDF assuming a uniform prior PDF and show that substantial differences between the two methods exist. In addition, we use the posterior PDF formalism, which was extended to include an unknown refractive index, to find credible sets for a variety of optical measurement scenarios. We find the uncertainty is greatly reduced with the addition of suitable extinction measurements in contrast to the inclusion of extra backscatter coefficients, which we show to have a minimal effect and strengthens similar observations based on numerical regularization methods. PMID- 18382594 TI - Laterally graded porous silicon optical filter fabricated by diffusion-limited etch process. AB - We report a method of producing a lateral gradient in the optical properties of anodically etched porous silicon layers. Lateral gradation details of the porous silicon layer are governed by the etch mask pattern involved. Unlike other methods that rely on uneven hole current distribution, we believe that in our method the diffusion of reactive ions in the etchant plays a key role. As an implementation of the proposed method, we demonstrate a linearly graded optical bandpass filter operating at the lambda=1550 nm range by employing a tapered etch window opening. The resultant optical filter exhibited a approximate 60 nm tuning range with a sharp transmission bandwidth of approximately 3 nm. Computer simulations indicate that an uneven hole current distribution cannot be the reason for the observed gradient along the taper axis, supporting the view that the diffusion-limited etch process plays the key role. PMID- 18382595 TI - Ultrafast dynamics of the real and imaginary permittivity parts of a photoexcited silver layer revealed by surface plasmon resonance. AB - Dynamics of the real and imaginary parts of the dielectric permittivity of a photoexcited silver layer has been investigated by means of femtosecond pump probe spectroscopy in a surface plasmon resonance Kretschmann configuration. Both real and imaginary permittivity parts experience changes in the visible-near-IR regions under silver excitation at 400 nm. The changes are stronger, particularly those of the imaginary part, at longer wavelengths. Three excited states are formed during the relaxation process, which are attributed to the nonequilibrium and equilibrium heated electron distributions and lattice heating. Different time evolutions of the real and imaginary dielectric permittivity parts are explained by different contributions from interband and intraband transitions caused by plasma frequency and electron scattering frequency variations. PMID- 18382597 TI - III-posed retrieval of aerosol extinction coefficient profiles from Raman lidar data by regularization. AB - In the analysis of Raman lidar measurements of aerosol extinction, it is necessary to calculate the derivative of the logarithm of the ratio between the atmospheric number density and the range-corrected lidar-received power. The statistical fluctuations of the Raman signal can produce large fluctuations in the derivative and thus in the aerosol extinction profile. To overcome this difficult situation we discuss three methods: Tikhonov regularization, variational, and the sliding best-fit (SBF). Three methods are performed on the profiles taken from the European Aerosol Research Lidar Network lidar database simulated at the Raman shifted wavelengths of 387 and 607 nm associated with the emitted signals at 355 and 532 nm. Our results show that the SBF method does not deliver good results for low fluctuation in the profile. However, Tikhonov regularization and the variational method yield very good aerosol extinction coefficient profiles for our examples. With regard to, e.g., the 532 nm wavelength, the L2 errors of the aerosol extinction coefficient profile by using the SBF, Tikhonov, and variational methods with respect to synthetic noisy data are 0.0015(0.0024), 0.00049(0.00086), and 0.00048(0.00082), respectively. Moreover, the L2 errors by using the Tikhonov and variational methods with respect to a more realistic noisy profile are 0.0014(0.0016) and 0.0012(0.0016), respectively. In both cases the L2 error given in parentheses concerns the second example. PMID- 18382598 TI - Refractive index variation in compression molding of precision glass optical components. AB - Compression molding of glass optical components is a high volume near net-shape precision fabrication method. In a compression molding process, a variation of the refractive index occurs along the radial direction of the glass component due to thermal treatment. The variation of refractive index is an important parameter that can affect the performance of optical lenses, especially lenses used for high precision optical systems. Refractive index variations in molded glass lenses under different cooling conditions were investigated using both an experimental approach and a numerical simulation. Specifically, refractive index variations inside molded glass lenses were evaluated by measuring optical wavefront variations with a Shack-Hartmann sensor system. The measured refractive index variations of the molded glass lenses were compared with the numerical simulation as a validation of the modeling approach. PMID- 18382599 TI - Simultaneous measurement for strain and temperature using fiber Bragg gratings and multimode fibers. AB - An all-fiber sensor capable of simultaneous measurement of temperature and strain is newly presented. The sensing head is formed by a fiber Bragg grating combined with a section of multimode fiber that acts as a Mach-Zehnder interferometer for temperature and strain discrimination. The strain and temperature coefficients of multimode fibers vary with the core sizes and materials. This feature can be used to improve the strain and temperature resolution by suitably choosing the multimode fiber. For a 10 pm wavelength resolution, a resolution of 9.21 mu epsilon in strain and 0.26 degrees C in temperature can be achieved. PMID- 18382600 TI - Camera for coherent diffractive imaging and holography with a soft-x-ray free electron laser. AB - We describe a camera to record coherent scattering patterns with a soft-x-ray free-electron laser (FEL). The camera consists of a laterally graded multilayer mirror, which reflects the diffraction pattern onto a CCD detector. The mirror acts as a bandpass filter for both the wavelength and the angle, which isolates the desired scattering pattern from nonsample scattering or incoherent emission from the sample. The mirror also solves the particular problem of the extreme intensity of the FEL pulses, which are focused to greater than 10(14) W/cm2. The strong undiffracted pulse passes through a hole in the mirror and propagates onto a beam dump at a distance behind the instrument rather than interacting with a beam stop placed near the CCD. The camera concept is extendable for the full range of the fundamental wavelength of the free electron laser in Hamburg (FLASH) FEL (i.e., between 6 and 60 nm) and into the water window. We have fabricated and tested various multilayer mirrors for wavelengths of 32, 16, 13.5, and 4.5 nm. At the shorter wavelengths mirror roughness must be minimized to reduce scattering from the mirror. We have recorded over 30,000 diffraction patterns at the FLASH FEL with no observable mirror damage or degradation of performance. PMID- 18382601 TI - Quantitative phase measurements using optical quadrature microscopy. AB - Imaging of phase or optical path length is becoming more important with the development of better imaging systems, computational algorithms, faster computers, and a greater interest in the imaging of transparent objects. Early phase imaging involved qualitative imaging of phase gradients. New computational algorithms can be used to extract some quantitative phase imaging from these techniques. In contrast, new hardware has enabled full-field quantitative phase imaging on a practical and cost-effective scale. We explore a quantitative comparison between two techniques for imaging phase. In the first technique, phase is recovered from a pair of differential interference contrast images, and in the second technique, phase is measured pixel-by-pixel interferometrically. It is shown, experimentally, that the overall results are similar, but each technique has its own advantages and disadvantages. PMID- 18382602 TI - Synthesis of alkylidenephthalans through fluoride-induced cyclization of electron deficient 2-siloxymethylphenylacetylene derivatives. AB - Fluoride-based deprotection of silylated 2-alkynylbenzyl alcohol derivatives featuring carbonyl-substituted alkynes results in the direct synthesis of alkylidenephthalan vinylogous esters. The reaction is selective for the Z alkylidenephthalans in a thermodynamically controlled process. Similar compounds are also produced in the coupling of Fischer carbene complexes with 2 alkynylbenzoyl derivatives in an aqueous solvent system. Subsequent acid catalyzed inter- or intramolecular Diels-Alder reactions lead to hydronaphthalene or hydrophenanthrene derivatives. PMID- 18382603 TI - NML computation algorithms for tree-structured multinomial Bayesian networks. AB - Typical problems in bioinformatics involve large discrete datasets. Therefore, in order to apply statistical methods in such domains, it is important to develop efficient algorithms suitable for discrete data. The minimum description length (MDL) principle is a theoretically well-founded, general framework for performing statistical inference. The mathematical formalization of MDL is based on the normalized maximum likelihood (NML) distribution, which has several desirable theoretical properties. In the case of discrete data, straightforward computation of the NML distribution requires exponential time with respect to the sample size, since the definition involves a sum over all the possible data samples of a fixed size. In this paper, we first review some existing algorithms for efficient NML computation in the case of multinomial and naive Bayes model families. Then we proceed by extending these algorithms to more complex, tree-structured Bayesian networks. PMID- 18382604 TI - Early diagnosis of neuropathy in leprosy--comparing diagnostic tests in a large prospective study (the INFIR cohort study). AB - BACKGROUND: Leprosy is the most frequent treatable neuromuscular disease. Yet, every year, thousands of patients develop permanent peripheral nerve damage as a result of leprosy. Since early detection and treatment of neuropathy in leprosy has strong preventive potential, we conducted a cohort study to determine which test detects this neuropathy earliest. METHODS AND FINDINGS: One hundred and eighty-eight multibacillary (MB) leprosy patients were selected from a cohort of 303 and followed for 2 years after diagnosis. Nerve function was evaluated at each visit using nerve conduction (NC), quantitative thermal sensory testing and vibrometry, dynamometry, monofilament testing (MFT), and voluntary muscle testing (VMT). Study outcomes were sensory and motor impairment detected by MFT or VMT. Seventy-four of 188 patients (39%) had a reaction, neuritis, or new nerve function impairment (NFI) event during a 2-year follow-up. Sub-clinical neuropathy was extensive (20%-50%), even in patients who did not develop an outcome event. Sensory nerve action potential (SNAP) amplitudes, compound motor action potential (CMAP) velocities, and warm detection thresholds (WDT) were most frequently affected, with SNAP impairment frequencies ranging from 30% (median) to 69% (sural). Velocity was impaired in up to 43% of motor nerves. WDTs were more frequently affected than cold detection thresholds (29% versus 13%, ulnar nerve). Impairment of SNC and warm perception often preceded deterioration in MF or VMT scores by 12 weeks or more. CONCLUSIONS: A large proportion of leprosy patients have subclinical neuropathy that was not evident when only MFT and VMT were used. SNC was the most frequently and earliest affected test, closely followed by WDT. They are promising tests for improving early detection of neuropathy, as they often became abnormal 12 weeks or more before an abnormal monofilament test. Changes in MFT and VMT score mirrored changes in neurophysiology, confirming their validity as screening tests. PMID- 18382605 TI - Molecular genetics reveal that silvatic Rhodnius prolixus do colonise rural houses. AB - BACKGROUND: Rhodnius prolixus is the main vector of Chagas disease in Venezuela. Here, domestic infestations of poor quality rural housing have persisted despite four decades of vector control. This is in contrast to the Southern Cone region of South America, where the main vector, Triatoma infestans, has been eliminated over large areas. The repeated colonisation of houses by silvatic populations of R. prolixus potentially explains the control difficulties. However, controversy surrounds the existence of silvatic R. prolixus: it has been suggested that all silvatic populations are in fact Rhodnius robustus, a related species of minor epidemiological importance. Here we investigate, by direct sequencing (mtcytb, D2) and by microsatellite analysis, 1) the identity of silvatic Rhodnius and 2) whether silvatic populations of Rhodnius are isolated from domestic populations. METHODS AND FINDINGS: Direct sequencing confirmed the presence of R. prolixus in palms and that silvatic bugs can colonise houses, with house and palm specimens sharing seven cytb haplotypes. Additionally, mitochondrial introgression was detected between R. robustus and R. prolixus, indicating a previous hybridisation event. The use of ten polymorphic microsatellite loci revealed a lack of genetic structure between silvatic and domestic ecotopes (non-significant F(ST) values), which is indicative of unrestricted gene flow. CONCLUSIONS: Our analyses demonstrate that silvatic R. prolixus presents an unquestionable threat to the control of Chagas disease in Venezuela. The design of improved control strategies is essential for successful long term control and could include modified spraying and surveillance practices, together with housing improvements. PMID- 18382606 TI - Human leptospirosis caused by a new, antigenically unique Leptospira associated with a Rattus species reservoir in the Peruvian Amazon. AB - As part of a prospective study of leptospirosis and biodiversity of Leptospira in the Peruvian Amazon, a new Leptospira species was isolated from humans with acute febrile illness. Field trapping identified this leptospire in peridomestic rats (Rattus norvegicus, six isolates; R. rattus, two isolates) obtained in urban, peri-urban, and rural areas of the Iquitos region. Novelty of this species was proven by serological typing, 16S ribosomal RNA gene sequencing, pulsed-field gel electrophoresis, and DNA-DNA hybridization analysis. We have named this species "Leptospira licerasiae" serovar Varillal, and have determined that it is phylogenetically related to, but genetically distinct from, other intermediate Leptospira such as L. fainei and L. inadai. The type strain is serovar Varillal strain VAR 010(T), which has been deposited into internationally accessible culture collections. By microscopic agglutination test, "Leptospira licerasiae" serovar Varillal was antigenically distinct from all known serogroups of Leptospira except for low level cross-reaction with rabbit anti-L. fainei serovar Hurstbridge at a titer of 1:100. LipL32, although not detectable by PCR, was detectable in "Leptospira licerasiae" serovar Varillal by both Southern blot hybridization and Western immunoblot, although on immunoblot, the predicted protein was significantly smaller (27 kDa) than that of L. interrogans and L. kirschneri (32 kDa). Isolation was rare from humans (2/45 Leptospira isolates from 881 febrile patients sampled), but high titers of MAT antibodies against "Leptospira licerasiae" serovar Varillal were common (30%) among patients fulfilling serological criteria for acute leptospirosis in the Iquitos region, and uncommon (7%) elsewhere in Peru. This new leptospiral species reflects Amazonian biodiversity and has evolved to become an important cause of leptospirosis in the Peruvian Amazon. PMID- 18382607 TI - An analysis of genetic diversity and inbreeding in Wuchereria bancrofti: implications for the spread and detection of drug resistance. AB - Estimates of genetic diversity in helminth infections of humans often have to rely on genotyping (immature) parasite transmission stages instead of adult worms. Here we analyse the results of one such study investigating a single polymorphic locus (a change at position 200 of the beta-tubulin gene) in microfilariae of the lymphatic filarial parasite Wuchereria bancrofti. The presence of this genetic change has been implicated in benzimidazole resistance in parasitic nematodes of farmed ruminants. Microfilariae were obtained from patients of three West African villages, two of which were sampled prior to the introduction of mass drug administration. An individual-based stochastic model was developed showing that a wide range of allele frequencies in the adult worm populations could have generated the observed microfilarial genetic diversity. This suggests that appropriate theoretical null models are required in order to interpret studies that genotype transmission stages. Wright's hierarchical F statistic was used to investigate the population structure in W. bancrofti microfilariae and showed significant deficiency of heterozygotes compared to the Hardy-Weinberg equilibrium; this may be partially caused by a high degree of parasite genetic differentiation between hosts. Studies seeking to quantify accurately the genetic diversity of helminth populations by analysing transmission stages should increase their sample size to account for the variability in allele frequency between different parasite life-stages. Helminth genetic differentiation between hosts and non-random mating will also increase the number of hosts (and the number of samples per host) that need to be genotyped, and could enhance the rate of spread of anthelmintic resistance. PMID- 18382608 TI - The Mathematical Foundations of 3D Compton Scatter Emission Imaging. AB - The mathematical principles of tomographic imaging using detected (unscattered) X or gamma-rays are based on the two-dimensional Radon transform and many of its variants. In this paper, we show that two new generalizations, called conical Radon transforms, are related to three-dimensional imaging processes based on detected Compton scattered radiation. The first class of conical Radon transform has been introduced recently to support imaging principles of collimated detector systems. The second class is new and is closely related to the Compton camera imaging principles and invertible under special conditions. As they are poised to play a major role in future designs of biomedical imaging systems, we present an account of their most important properties which may be relevant for active researchers in the field. PMID- 18382609 TI - Myeloperoxidase: a new biomarker of inflammation in ischemic heart disease and acute coronary syndromes. AB - Myeloperoxidase (MPO) is an enzyme stored in azurophilic granules of polymorphonuclear neutrophils and macrophages and released into extracellular fluid in the setting of inflammatory process. The observation that myeloperoxidase is involved in oxidative stress and inflammation has been a leading factor to study myeloperoxidase as a possible marker of plaque instability and a useful clinical tool in the evaluation of patients with coronary heart disease. The purpose of this review is to provide an overview of the pathophysiological, analytical, and clinical characteristics of MPO and to summarize the state of art about the possible clinical use of MPO as a marker for diagnosis and risk stratification of patients with acute coronary syndrome (ACS). PMID- 18382610 TI - Coexistence of two rare sarcomas: primary leiomyosarcoma of bone and epithelioid hemangioendothelioma of the liver. AB - A 33-year-old woman sought medical attention for a painful swelling of the left ankle. Plain radiographs revealed an osteolytic lesion involving the left distal tibia. An excisional biopsy provided the diagnosis of leiomyosarcoma in the tibia. A staging work-up was performed and an abdominal CT showed 4 liver hypodense lesions in both lobes with peripheral contrast enhancement. A liver biopsy confirmed the diagnosis of epithelioid hemangioendothelioma of the liver. No association between these two entities has been described before. This case introduces the importance of the pathological confirmation of apparent metastatic lesions in low grade sarcomas and provides a review of the literature of both tumours. PMID- 18382611 TI - Elastofibroma dorsi: an uncommon benign pseudotumour. AB - Elastofibroma dorsi is an uncommon benign soft tissue pseudotumour usually located at the lower pole of the scapula, deep to serratus anterior, and often attached to the periosteum of the ribs, presenting with long history of swelling and occasionally pain and discomfort. This lesion is usually seen in patients over the age of 50 years and is not uncommonly mistaken as a malignant tumour because of its size and location deep to the periscapular muscles. Review of the orthopaedic oncology database of 17 500 patients revealed that there were 15 patients with elastofibroma dorsi. There were 12 males and 3 females, mean age at diagnosis of 68.4 years range 51-79 years. The diagnosis was confirmed by MRI in 3 patients, excision biopsy in 3 patients, trucut biopsy in 8 patients and open biopsy in 1 patient. Eight patients had excision of the lesion which was symptomatic. There have been no recurrences. We highlight the clinical and radiological presentation of elastofibroma dorsi to increase awareness of its existence and management. PMID- 18382612 TI - A Role for PPARbeta/delta in Ocular Angiogenesis. AB - The uses of highly selective PPARbeta/delta ligands and PPARbeta/delta knockout mice have shown a direct ability of PPARbeta/delta to regulate angiogenesis in vitro and in vivo in animal models. PPARbeta/delta ligands induce the proangiogenic growth factor VEGF in many cells and tissues, though its actions in the eye are not known. However, virtually, all tissue components of the eye express PPARbeta/delta. Both angiogenesis and in particular VEGF are not only critical for the development of the retina, but they are also a central component in many common pathologies of the eye, including diabetic retinopathy and age related macular degeneration, the most common causes of blindness in the Western world. This review, therefore, will discuss the recent evidence of PPARbeta/delta mediated angiogenesis and VEGF release in the context of ocular disorders. PMID- 18382613 TI - Effects of radix adenophorae and cyclosporine A on an OVA-induced murine model of asthma by suppressing to T cells activity, eosinophilia, and bronchial hyperresponsiveness. AB - The present study is performed to investigate the inhibitory effects of Radix Adenophorae extract (RAE) on ovalbumin-induced asthma murine model. To study the anti-inflammatory and antiasthmatic effects of RAE, we examined the development of pulmonary eosinophilic inflammation and inhibitory effects of T cells in murine by RAE and cyclosporine A (CsA). We examined determination of airway hyperresponsiveness, flow cytometric analysis (FACS), enzyme-linked immunosorbent assay (ELISA), quantitative real time (PCR), hematoxylin-eosin staining, and Masson trichrome staining in lung tissue, lung weight, total cells, and eosinophil numbers in lung tissue. We demonstrated how RAE suppressed development on inflammation and decreased airway damage. PMID- 18382614 TI - Pulmonary tuberculosis in a young pregnant female: challenges in diagnosis and management. AB - BACKGROUND: With the world becoming a global village, tuberculosis is no longer limited to endemic areas. Our case emphasizes the impact of immigration on infectious disease epidemiology and challenges associated with diagnosis and treatment in pregnancy. CASE: A 21-year-old Hispanic female presented in preterm labor and was found to be hypoxic. Chest X-ray revealed a paratracheal mass which a CT scan confirmed. PPD test was positive. Bronchoalveolar lavage did not reveal acid-fast bacilli and biopsy revealed caseating granulomas. Diagnosis and treatment were challenging due to constraints in radiological investigations, lack of initial evidence of acid-fast bacilli, and toxic profile of medications. Due to her high risk, she was started on antituberculosis regimen. The diagnosis was confirmed on Day 26 when Mycobacterium tuberculosis was isolated by DNA probe. CONCLUSION: A high index of suspicion is required to recognize the changing face and disease spectrum of tuberculosis and initiate treatment for better outcomes. PMID- 18382615 TI - Barley genomics: An overview. AB - Barley (Hordeum vulgare), first domesticated in the Near East, is a well-studied crop in terms of genetics, genomics, and breeding and qualifies as a model plant for Triticeae research. Recent advances made in barley genomics mainly include the following: (i) rapid accumulation of EST sequence data, (ii) growing number of studies on transcriptome, proteome, and metabolome, (iii) new modeling techniques, (iv) availability of genome-wide knockout collections as well as efficient transformation techniques, and (v) the recently started genome sequencing effort. These developments pave the way for a comprehensive functional analysis and understanding of gene expression networks linked to agronomically important traits. Here, we selectively review important technological developments in barley genomics and related fields and discuss the relevance for understanding genotype-phenotype relationships by using approaches such as genetical genomics and association studies. High-throughput genotyping platforms that have recently become available will allow the construction of high-density genetic maps that will further promote marker-assisted selection as well as physical map construction. Systems biology approaches will further enhance our knowledge and largely increase our abilities to design refined breeding strategies on the basis of detailed molecular physiological knowledge. PMID- 18382617 TI - Use of recombinant antigens for the diagnosis of invasive candidiasis. AB - Invasive candidiasis is a frequent and often fatal complication in immunocompromised and critically ill patients. Unfortunately, the diagnosis of invasive candidiasis remains difficult due to the lack of specific clinical symptoms and a definitive diagnostic method. The detection of antibodies against different Candida antigens may help in the diagnosis. However, the methods traditionally used for the detection of antibodies have been based on crude antigenic fungal extracts, which usually show low-reproducibility and cross reactivity problems. The development of molecular biology techniques has allowed the production of recombinant antigens which may help to solve these problems. In this review we will discuss the usefulness of recombinant antigens in the diagnosis of invasive candidiasis. PMID- 18382616 TI - PPAR-gamma, Microglial Cells, and Ocular Inflammation: New Venues for Potential Therapeutic Approaches. AB - The last decade has witnessed an increasing interest for the role played by the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) in controlling inflammation in peripheral organs as well as in the brain. Activation of PPAR gamma has been shown to control the response of microglial cells, the main macrophage population found in brain parenchyma, and limit the inflammation. The anti-inflammatory capacity of PPAR-gamma agonists has led to the hypothesis that PPAR-gamma might be targeted to modulate degenerative brain diseases in which inflammation has been increasingly recognized as a significant component. Recent experimental evidence suggests that PPAR-gamma agonists could be exploited to treat ocular diseases such as diabetic retinopathy, age-related macular degeneration, autoimmune uveitis, and optic neuritis where inflammation has relevant role. Additional PPAR-gamma agonist beneficial effects could involve amelioration of retinal microcirculation and inhibition of neovascularization. However, PPAR-gamma activation could, in some instances, aggravate the ocular pathology, for example, by increasing the synthesis of vascular endothelial growth factor, a proangiogenic factor that could trigger a vicious circle and further deteriorate retinal perfusion. The development of new in vivo and in vitro models to study ocular inflammation and how to modulate for the eye benefit will be instrumental for the search of effective therapies. PMID- 18382618 TI - Intracellular signalling by C-peptide. AB - C-peptide, a cleavage product of the proinsulin molecule, has long been regarded as biologically inert, serving merely as a surrogate marker for insulin release. Recent findings demonstrate both a physiological and protective role of C-peptide when administered to individuals with type I diabetes. Data indicate that C peptide appears to bind in nanomolar concentrations to a cell surface receptor which is most likely to be G-protein coupled. Binding of C-peptide initiates multiple cellular effects, evoking a rise in intracellular calcium, increased PI 3-kinase activity, stimulation of the Na(+)/K(+) ATPase, increased eNOS transcription, and activation of the MAPK signalling pathway. These cell signalling effects have been studied in multiple cell types from multiple tissues. Overall these observations raise the possibility that C-peptide may serve as a potential therapeutic agent for the treatment or prevention of long term complications associated with diabetes. PMID- 18382619 TI - Peroxisome proliferator-activated receptors: "key" regulators of neuroinflammation after traumatic brain injury. AB - Traumatic brain injury is characterized by neuroinflammatory pathological sequelae which contribute to brain edema and delayed neuronal cell death. Until present, no specific pharmacological compound has been found, which attenuates these pathophysiological events and improves the outcome after head injury. Recent experimental studies suggest that targeting peroxisome proliferator activated receptors (PPARs) may represent a new anti-inflammatory therapeutic concept for traumatic brain injury. PPARs are "key" transcription factors which inhibit NFkappaB activity and downstream transcription products, such as proinflammatory and proapoptotic cytokines. The present review outlines our current understanding of PPAR-mediated neuroprotective mechanisms in the injured brain and discusses potential future anti-inflammatory strategies for head injured patients, with an emphasis on the putative beneficial combination therapy of synthetic cannabinoids (e.g., dexanabinol) with PPARalpha agonists (e.g., fenofibrate). PMID- 18382620 TI - Structure-based inhibitors exhibit differential activities against Helicobacter pylori and Escherichia coli undecaprenyl pyrophosphate synthases. AB - Helicobacter pylori colonizes the human gastric epithelium and causes diseases such as gastritis, peptic ulcers, and stomach cancer. Undecaprenyl pyrophosphate synthase (UPPS), which catalyzes consecutive condensation reactions of farnesyl pyrophosphate with eight isopentenyl pyrophosphate to form lipid carrier for bacterial peptidoglycan biosynthesis, represents a potential target for developing new antibiotics. In this study, we solved the crystal structure of H. pylori UPPS and performed virtual screening of inhibitors from a library of 58,635 compounds. Two hits were found to exhibit differential activities against Helicobacter pylori and Escherichia coli UPPS, giving the possibility of developing antibiotics specially targeting pathogenic H. pylori without killing the intestinal E. coli. PMID- 18382621 TI - Prevention of Oxidative Stress-Induced Retinal Pigment Epithelial Cell Death by the PPARgamma Agonists, 15-Deoxy-Delta 12, 14-Prostaglandin J(2). AB - Cellular oxidative stress plays an important role in retinal pigment epithelial (RPE) cell death during aging and the development of age-related macular degeneration. Early reports indicate that during phagocytosis of rod outer segments, there is an increase of RPE oxidative stress and an upregulation of PPARgamma mRNA in these cells. These studies suggest that activation of PPARgamma may modulate cellular oxidative stress. This paper presents a brief review of recent studies that investigate RPE oxidative stress under various experimental conditions. This is followed by a detailed review on those reports that examine the protective effect of the natural PPARgamma ligand, 15d-PGJ(2), against RPE oxidative stress. This agent can upregulate glutathione and prevent oxidant induced intracellular reactive oxygen species accumulation, mitochondrial depolarization, and apoptosis. The cytoprotective effect of this agent, however, is not shared by other PPARgamma agonists. Nonetheless, this property of 15d PGJ(2) may be useful in future development of pharmacological tools against retinal diseases caused by oxidative stress. PMID- 18382622 TI - Adult head and neck soft tissue sarcomas: treatment and outcome. AB - We have retrospectively analysed the experience of a musculoskeletal oncological unit in the management of adult head and neck soft tissue sarcomas from 1990 to 2005. Thirty-six patients were seen, of whom 24 were treated at this unit, the remainder only receiving advice. The median age of the patients was 46 years. Most of the sarcomas were deep and of high or intermediate grade with a median size of 5.5 cm. Eleven different histological subtypes were identified. Wide excision was possible only in 21% of the cases. 42% of the patients developed local recurrence and 42% developed metastatic disease usually in the lungs. Overall survival was 49% at 5 years. Tumour size was the most important prognostic factor. Adult head and neck soft tissue sarcomas have a high mortality rate with a high risk of local recurrence and metastatic disease. The rarity of the disease would suggest that centralisation of care could lead to increased expertise and better outcomes. PMID- 18382623 TI - MMSE reconstruction for 3D freehand ultrasound imaging. AB - The reconstruction of 3D ultrasound (US) images from mechanically registered, but otherwise irregularly positioned, B-scan slices is of great interest in image guided therapy procedures. Conventional 3D ultrasound algorithms have low computational complexity, but the reconstructed volume suffers from severe speckle contamination. Furthermore, the current method cannot reconstruct uniform high-resolution data from several low-resolution B-scans. In this paper, the minimum mean-squared error (MMSE) method is applied to 3D ultrasound reconstruction. Data redundancies due to overlapping samples as well as correlation of the target and speckle are naturally accounted for in the MMSE reconstruction algorithm. Thus, the reconstruction process unifies the interpolation and spatial compounding. Simulation results for synthetic US images are presented to demonstrate the excellent reconstruction. PMID- 18382624 TI - Analysis of gene expression in normal and cancer cells exposed to gamma radiation. AB - The expression of many genes is modulated after exposure to ionizing radiation. Identification of specific genes may allow the determination of pathways important in radiation responses. We previously identified modulation of the expression of several genes in response to ionizing radiation treatment. In the present study, we monitored the expression of RGS1, CC3, THBS1, vWF, MADH7, and a novel gene encoding a secreted protein in irradiated Jurkat, TK6, HeLa, and HFL1 cells. The RGS1 is involved in G-protein signaling pathway, CC3 belongs to the complement system, THBS1 is a component of the extracellular matrix, vWF takes part in blood coagulation, and MADH7 is a member of the TGF-beta signal transduction pathway. Our objective was to find similarities and differences in the expression of these genes in ionizing radiation-exposed diverse cell types. RGS1 was downregulated in Jurkat cells but was upregulated in TK6 and HFL1 cells. The expression of CC3 was repressed in Jurkat and HFL1 cells but was induced in TK6 and HeLa cells. THBS1 was downregulated in irradiated TK6 and HFL1 cells. vWF was induced in radiation-exposed HeLa cells, but its expression was downregulated in Jurkat cells. The expression of MADH7 was induced in all the cell types examined. These results indicate cell specific modulation of gene expression and suggest the involvement of different pathways in cellular response to radiation treatment in different cells. PMID- 18382625 TI - Design of biotin-functionalized luminescent quantum dots. AB - We report the design and synthesis of a tetraethylene glycol- (TEG-) based bidentate ligand functionalized with dihydrolipoic acid (DHLA) and biotin (DHLA TEG-biotin) to promote biocompatibility of luminescent quantum dots (QD's). This new ligand readily binds to CdSe-ZnS core-shell QDs via surface ligand exchange. QDs capped with a mixture of DHLA and DHLA-TEG-biotin or polyethylene glycol- (PEG-) (molecular weight average approximately 600) modified DHLA (DHLA-PEG600) and DHLA-TEG-biotin are easily dispersed in aqueous buffer solutions. In particular, homogeneous buffer solutions of QDs capped with a mixture of DHLA PEG600 and DHLA-TEG-biotin that are stable over broad pH range have been prepared. QDs coated with mixtures of DHLA/DHLA-TEG-biotin and with DHLA PEG600/DHLA-TEG-biotin were tested in surface binding assays and the results indicate that biotin groups on the QD surface interact specifically with NeutrAvidin-functionalized microtiter well plates. PMID- 18382627 TI - Planning for the pandemic. PMID- 18382626 TI - Dedifferentiated Liposarcoma of the Retroperitoneum with Extensive Leiomyosarcomatous Differentiation and beta-Human Chorionic Gonadotropin Production. AB - Dedifferentiated liposarcomas may display a variety of "heterologous" lines of differentiation, including osseous, vascular, skeletal, and/or smooth muscular. There have been six previously reported examples of leiomyosarcomas associated with high levels of serum human chorionic gonadotropin (hCG) production, comprised of cases originating from the retroperitoneum, spermatic cord, small intestine, and uterus. This report describes the first example of a dedifferentiated liposarcoma that combined both of the aforementioned features: extensive heterologous (leiomyosarcomatous) differentiation and beta-hCG production (maximum serum levels 1046 mIU/ml, reference <5 mIU/ml). The tumor, which originated in the retroperitoneum in the region of the right kidney, was rapidly progressive and ultimately fatal within three months of its diagnosis. In addition to characteristic morphologic features, lipogenic and smooth muscle differentiation were confirmed with immunohistochemical stains for MDM2 and smooth muscle actin, respectively. The tumor also displayed diffuse immunoreactivity for beta-hCG in both primary and metastatic sites. This case further expands the clinicopathologic spectrum of lipogenic tumors. PMID- 18382629 TI - Pandemic Influenza and Canada's Children. PMID- 18382628 TI - Tumour necrosis factor inhibitors and infection: What is there to know for infectious diseases physicians? PMID- 18382630 TI - Infectious disease management: lessons from cuba. PMID- 18382631 TI - A comparison between the Strep A Rapid Test Device and conventional culture for the diagnosis of streptococcal pharyngitis. AB - BACKGROUND: Rapid antigen detection tests are frequently used to diagnose pharyngitis due to Streptococcus pyogenes. Because a large number of kits are available commercially, performance characteristics may vary considerably. The present study evaluated one such kit currently in use in Canadian laboratories for which published evaluations are not available. OBJECTIVE: To evaluate the performance characteristics of the Strep A Rapid Test Device (SARTD) (Nova Century Scientific Inc, Canada). METHODS: Pharyngeal swabs from 818 patients with suspected streptococcal pharyngitis were tested. Swabs were initially inoculated onto the surface of a blood agar plate and then used to perform the rapid antigen test. The test was performed in accordance with the product monograph. Beta hemolytic colonies were identified as S pyogenes using conventional means. RESULTS: Four hundred ninety specimens were obtained from children and 328 from adults. S pyogenes was recovered from 171 (21%) patients. The SARTD detected S pyogenes antigens in 123 of 171 specimens from which S pyogenes was isolated on culture; the screen was negative in 610 of 647 specimens from which cultures were negative. The positive and negative predictive values of the SARTD were 76.9% and 92.7%, respectively. CONCLUSIONS: The SARTD was much less sensitive (72%) than was suggested in the product monograph (90%). Laboratories should vigorously evaluate such products in-house, optimize specimen collection and transport, and choose more sensitive kits for use. PMID- 18382632 TI - Prenatal Screening for HIV in Nova Scotia: Survey of Postpartum Women and Audit of Current Prenatal Screening Practices. AB - BACKGROUND: Current guidelines for screening for HIV infections in Nova Scotia recommend an opt-in approach in which patients are counselled and consent to testing. The objectives of the present study were to measure adherence to these recommendations, to explore women's knowledge, attitudes, beliefs and behaviours concerning HIV screening, and to compare these results with prenatal screening practices for rubella, hepatitis B and group B streptococcus. METHODS: All women who gave birth consecutively during a seven-week period were recruited. Study participants were interviewed to determine their knowledge, attitudes and beliefs concerning prenatal screening. Hospital and laboratory records were reviewed for information concerning prenatal screening and perinatal treatment to audit screening practices. RESULTS: A total of 279 patients were enrolled in the study, representing 58% of those eligible. The HIV screening rate was 72%, compared with 95% for rubella, 89% for hepatitis B and 24% for group B streptococcus. Of the participants tested for HIV, 80% were aware of being tested. Of all the study participants, 17% indicated having received pretest counselling about HIV, 56% volunteered to be tested for HIV, 78% received the test results, and 3.8% received post-test counselling. More participants preferred an opt-out approach to HIV screening (50%), where testing is routinely performed on everyone, rather than the opt-in approach (43%). Participants displayed a similar preference for screening for the other infections. INTERPRETATION: HIV prenatal testing rates in Nova Scotia are comparable with those of other provinces that recommend an opt-in approach, but are lower than testing rates for opt-out programs. Most study participants were not screened using the recommended opt-in approach. PMID- 18382633 TI - Estimated Numbers of Community Cases of Illness Due to Salmonella, Campylobacter and Verotoxigenic Escherichia Coli: Pathogen-specific Community Rates. AB - OBJECTIVE: To estimate the annual number of cases of illness due to verotoxigenic Escherichia coli (VTEC), Salmonella and Campylobacter in the Canadian population, using data from the National Notifiable Disease registry (NND), estimates of under-reporting derived from several National Studies on Acute Gastrointestinal Illness, and the literature. METHODS: For each of the three pathogens (VTEC, Salmonella and Campylobacter), data were used to estimate the percentage of cases reported at each step in the surveillance system. The number of reported cases in the NND for each pathogen was then divided by these percentages. In cases where the pathogen-specific estimates were unavailable, data on acute gastrointestinal illness were used, accounting for differences between those with bloody and nonbloody diarrhea. RESULTS: For every case of VTEC, Salmonella and Campylobacter infection reported in the NND, there were an estimated 10 to 47, 13 to 37, and 23 to 49 cases annually in the Canadian population, respectively. CONCLUSIONS: The authors estimate that a significant number of infections due to VTEC, Salmonella and Campylobacter occur each year in Canada, highlighting the fact that these enteric pathogens still pose a significant health burden. Recognizing the significant amount of under-reporting is essential to designing appropriate interventions and assessing the impact of these pathogens in the population. PMID- 18382634 TI - Syndromic Surveillance of Norovirus using Over-the-counter Sales of Medications Related to Gastrointestinal Illness. AB - OBJECTIVE: To assess whether over-the-counter (OTC) sales of gastrointestinal illness (GI)-related medications are associated with temporal trends of reportable community viral, bacterial and parasitic infections. METHODS: The temporal patterns in weekly and seasonal sales of nonprescription products related to GI were compared with those of reportable viral, bacterial and parasitic infections in a Canadian province. RESULTS: Temporal patterns of OTC product sales and Norovirus activity were similar, both having highest activity in the winter months. In contrast, GI cases from both bacterial and parasitic agents were highest from late spring through to early fall. CONCLUSIONS: Nonprescription sales of antidiarrheal and antinauseant products are a good predictor of community Norovirus activity. Syndromic surveillance through monitoring of OTC product sales could be useful as an early indicator of the Norovirus season, allowing for appropriate interventions to reduce the number of infections. PMID- 18382635 TI - Antimicrobial resistance among Salmonella and Shigella isolates in five Canadian provinces (1997 to 2000). AB - OBJECTIVE: To describe rates of antimicrobial resistance (AMR) among Salmonella and Shigella isolates reported in five Canadian provinces, focusing on clinically important antimicrobials. METHODS: The authors retrospectively investigated AMR rates among 6219 Salmonella and 1673 Shigella isolates submitted to provincial public health laboratories in Alberta, Newfoundland and Labrador, Ontario, Prince Edward Island and Saskatchewan from 1997 to 2000; these isolates were estimated to represent 41% of Salmonella cases and 72% of Shigella cases reported by the study provinces. RESULTS: Among Salmonella isolates, 27% (1704 of 6215) were resistant to ampicillin, 2.2% (135 of 6122) to trimethoprim/ sulfamethoxazole, 1.5% (14 of 938) to nalidixic acid, 1.2% (one of 84) to lomafloxacin and 0.08% (five of 6163) to ciprofloxacin. Among Shigella isolates, 70% (1144 of 1643) were resistant to trimethoprim/sulfamethoxazole, 65% (1079 of 1672) to ampicillin, 3.1% (eight of 262) to nalidixic acid, 0.49% (eight of 1636) to ciprofloxacin, 0.14% (one of 700) to ceftriaxone and 0.08% (one of 1292) to ceftazidime. CONCLUSIONS: Higher rates of resistance to clinically important antimicrobials (including ciprofloxacin) were observed among both Salmonella and Shigella isolates than has previously been reported. Current Canadian data on rates of AMR for these pathogens are required. PMID- 18382636 TI - Beyond antibiotics? PMID- 18382637 TI - Colistin: the phoenix arises. PMID- 18382638 TI - Maternal infectious diseases, antimicrobial therapy or immunizations: very few contraindications to breastfeeding. PMID- 18382639 TI - The use of antiviral drugs for influenza: recommended guidelines for practitioners. AB - The present document outlines current guidelines and supporting literature relating to the use of antiviral drugs for chemoprophylaxis and influenza illness therapy in paediatric and adult settings. The focus is on the management of influenza in interpandemic periods. Where appropriate, the areas in need of additional research are identified. It will be necessary to update aspects of these guidelines as new information emerges. The recommendations that follow represent the results of a joint effort supported by the Canadian Paediatric Society and the Association of Medical Microbiology and Infectious Disease Canada. PMID- 18382640 TI - Desperate therapies and the amazing race for antimicrobial alternatives. PMID- 18382641 TI - Where have All the Antibiotics Gone? AB - The discovery of antibiotics some 60 years ago was anticipated to herald the end of infectious diseases. However, microbial evolution and genetic jugglery have dispelled this notion; the constant increase in the appearance of resistant strains has not been matched by the introduction of new therapeutic agents. On the contrary, the dire need for novel antibiotics has coincided with a reduction in antibiotic discovery programs in the pharmaceutical industry. As a result, the treatment of microbial diseases has reached a point where many infections are essentially untreatable by the antimicrobial agents currently available. At the present time, numerous initiatives are being undertaken by physicians and by governments in an attempt to redress this situation. In addition, alternative approaches to antibiotics for the treatment of infectious diseases are being explored intensively. PMID- 18382642 TI - Probiotics to prevent the need for, and augment the use of, antibiotics. AB - Although humans and microbes are inseparable, our knowledge and understanding of the majority of microbes that help keep us alive and well is in desperate need of further investigation. Of the organisms that influence humans before birth and inhabit various niches from birth to old age, we know little about their identity, origin, metabolic properties, attributes and mechanisms of interactions with the host and surrounding microbes. The use of probiotics ("live microorganisms which when administered in adequate amounts confer a health benefit on the host") has re-emerged as a means to restore and boost the beneficial microbes in our bodies. The timing of resurgent interest in this ancient field coincides with the need to augment or replace antibiotics whose side effects are unwelcome and whose efficacy is diminishing due to drug resistance. Evidence that probiotic strains can act as adjuncts to antibiotic therapy by reducing adverse effects, improving antibiotic function and enhancing mucosal immunity is mounting. It is to our discredit that basic research on microbial ecology has been stalled in Canada for the past 20 years. If supported, research into indigenous and probiotic microbes will form an important part of future research that sheds light on health, disease and a basic understanding of life itself. In some cases, probiotics will be the difference between a good quality of life and a bad one, or perhaps even life over death. Improvements in clinical studies, manufacturing and regulatory standards must coincide with this progress to ensure that physicians and consumers have reliable, proven products for safe and efficacious use. PMID- 18382645 TI - Learning from mistakes. PMID- 18382643 TI - Phage Therapy - Everything Old is New Again. AB - The study of bacterial viruses (bacteriophages or phages) proved pivotal in the nascence of the disciplines of molecular biology and microbial genetics, providing important information on the central processes of the bacterial cell (DNA replication, transcription and translation) and on how DNA can be transferred from one cell to another. As a result of the pioneering genetics studies and modern genomics, it is now known that phages have contributed to the evolution of the microbial cell and to its pathogenic potential. Because of their ability to transmit genes, phages have been exploited to develop cloning vector systems. They also provide a plethora of enzymes for the modern molecular biologist. Until the introduction of antibiotics, phages were used to treat bacterial infections (with variable success). Western science is now having to re evaluate the application of phage therapy - a therapeutic modality that never went out of vogue in Eastern Europe - because of the emergence of an alarming number of antibiotic-resistant bacteria. The present article introduces the reader to phage biology, and the benefits and pitfalls of phage therapy in humans and animals. PMID- 18382644 TI - Tweaking Innate Immunity: the Promise of Innate Immunologicals As Anti infectives. AB - New and exciting insights into the importance of the innate immune system are revolutionizing our understanding of immune defense against infections, pathogenesis, and the treatment and prevention of infectious diseases. The innate immune system uses multiple families of germline-encoded pattern recognition receptors (PRRs) to detect infection and trigger a variety of antimicrobial defense mechanisms. PRRs are evolutionarily highly conserved and serve to detect infection by recognizing pathogen-associated molecular patterns that are unique to microorganisms and essential for their survival. Toll-like receptors (TLRs) are transmembrane signalling receptors that activate gene expression programs that result in the production of proinflammatory cytokines and chemokines, type I interferons and antimicrobial factors. Furthermore, TLR activation facilitates and guides activation of adaptive immune responses through the activation of dendritic cells. TLRs are localized on the cell surface and in endosomal/lysosomal compartments, where they detect bacterial and viral infections. In contrast, nucleotide-binding oligomerization domain proteins and RNA helicases are located in the cell cytoplasm, where they serve as intracellular PRRs to detect cytoplasmic infections, particularly viruses. Due to their ability to enhance innate immune responses, novel strategies to use ligands, synthetic agonists or antagonists of PRRs (also known as 'innate immunologicals') can be used as stand-alone agents to provide immediate protection or treatment against bacterial, viral or parasitic infections. Furthermore, the newly appreciated importance of innate immunity in initiating and shaping adaptive immune responses is contributing to our understanding of vaccine adjuvants and promises to lead to improved next-generation vaccines. PMID- 18382646 TI - Alemtuzumab and natalizumab: the monoclonal antibody story continues. PMID- 18382647 TI - An Outbreak of Human Coronavirus OC43 Infection and Serological Cross-reactivity with SARS Coronavirus. AB - BACKGROUND: In summer 2003, a respiratory outbreak was investigated in British Columbia, during which nucleic acid tests and serology unexpectedly indicated reactivity for severe acute respiratory syndrome coronavirus (SARS-CoV). METHODS: Cases at a care facility were epidemiologically characterized and sequentially investigated for conventional agents of respiratory infection, SARS-CoV and other human CoVs. Serological cross-reactivity between SARS-CoV and human CoV-OC43 (HCoV-OC43) was investigated by peptide spot assay. RESULTS: Ninety-five of 142 residents (67%) and 53 of 160 staff members (33%) experienced symptoms of respiratory infection. Symptomatic residents experienced cough (66%), fever (21%) and pneumonia (12%). Eight residents died, six with pneumonia. No staff members developed pneumonia. Findings on reverse transcriptase-polymerase chain reaction assays for SARS-CoV at a national reference laboratory were suspected to represent false positives, but this was confounded by concurrent identification of antibody to N protein on serology. Subsequent testing by reverse transcriptase polymerase chain reaction confirmed HCoV-OC43 infection. Convalescent serology ruled out SARS. Notably, sera demonstrated cross-reactivity against nucleocapsid peptide sequences common to HCoV-OC43 and SARS-CoV. CONCLUSIONS: These findings underscore the virulence of human CoV-OC43 in elderly populations and confirm that cross-reactivity to antibody against nucleocapsid proteins from these viruses must be considered when interpreting serological tests for SARS-CoV. PMID- 18382648 TI - Community-acquired antibiotic resistance in urinary isolates from adult women in Canada. AB - BACKGROUND: There are currently limited data regarding the prevalence of antimicrobial-resistant organisms causing community-acquired urinary tract infections among adult women in Canada. Trimethoprim-sulfamethoxazole (TMP-SMX) is the recommended first-line empirical antibiotic treatment, unless resistance of Escherichia coli to TMP-SMX exceeds 20%. OBJECTIVE: To assess current levels of TMP-SMX-resistant E coli in community-acquired cases of urinary tract infection in adult women. METHOD: Assessment of urine culture reports obtained from 21 laboratories across Canada, submitted by family physicians for women aged 16 years and older. RESULTS: In 2199 adult women with a positive urine culture, 1079 (49.1%) of pathogens isolated were resistant to at least one antibiotic and 660 (30.0%) were multidrug-resistant (resistant to two or more antibiotics). TMP SMX resistance was seen in 245 of 1613 (15.2%) E coli isolates (95% CI 13.5 to 17.0). This proportion was higher in women 50 years of age and older (155 of 863 isolates [18.0%]; P=0.001), in British Columbia (70 of 342 isolates [20.5%]) and in Ontario (62 of 370 isolates [16.8%]) when compared with eastern provinces (65 of 572 isolates [11.4%]; P=0.001). Fluoroquinolone-resistant E coli occurred in 107 of 1557 (6.9%) isolates (95% CI 5.7 to 8.2), with the highest level found in British Columbia (54 of 341 isolates [15.8%]; P=0.001). CONCLUSION: TMP-SMX continues to be appropriate as first-line empirical treatment of acute cystitis in adult women in Canada, as resistance remains below 20%. However, TMP-SMX resistance is higher in older women and in some provinces. The level of fluoroquinolone-resistant E coli is highest in British Columbia. PMID- 18382649 TI - Q Fever Presenting As Recurrent, Culture-negative Endocarditis with Aortic Prosthetic Valve Failure: A Case Report and Review of the Literature. AB - The present report describes a case of recurrent, culture-negative endocarditis presenting with aortic prosthetic valve dysfunction in a 62-year-old man who required four valve replacement surgeries. On each occasion, he presented with valve failure. Fever was only documented during his first presentation. Furthermore, no vegetations were detectable on his aortic valve at transesophageal echocardiography. On the occasion of his most recent presentation, a detailed history of animal exposure - including hunting and skinning deer, moose and other large animals with his bare hands - was the only clue to his diagnosis. Serum antibodies against Coxiella burnetii were strongly positive, and C burnetii DNA was detected by polymerase chain reaction from his resected aortic valve tissue. Q fever is a worldwide zoonotic infection with diverse reservoirs. This diagnosis should be considered when evaluating unexplained prosthetic valve dysfunction, particularly in the setting of animal exposure. PMID- 18382650 TI - A patient with microcytic anemia and Fever. PMID- 18382651 TI - Cell death by SecTRAPs: thioredoxin reductase as a prooxidant killer of cells. AB - BACKGROUND: SecTRAPs (selenium compromised thioredoxin reductase-derived apoptotic proteins) can be formed from the selenoprotein thioredoxin reductase (TrxR) by targeting of its selenocysteine (Sec) residue with electrophiles, or by its removal through C-terminal truncation. SecTRAPs are devoid of thioredoxin reductase activity but can induce rapid cell death in cultured cancer cell lines by a gain of function. PRINCIPAL FINDINGS: Both human and rat SecTRAPs killed human A549 and HeLa cells. The cell death displayed both apoptotic and necrotic features. It did not require novel protein synthesis nor did it show extensive nuclear fragmentation, but it was attenuated by use of caspase inhibitors. The redox active disulfide/dithiol motif in the N-terminal domain of TrxR had to be maintained for manifestation of SecTRAP cytotoxicity. Stopped-flow kinetics showed that NADPH can reduce the FAD moiety in SecTRAPs at similar rates as in native TrxR and purified SecTRAPs could maintain NADPH oxidase activity, which was accelerated by low molecular weight substrates such as juglone. In a cellular context, SecTRAPs triggered extensive formation of reactive oxygen species (ROS) and consequently antioxidants could protect against the cell killing by SecTRAPs. CONCLUSIONS: We conclude that formation of SecTRAPs could contribute to the cytotoxicity seen upon exposure of cells to electrophilic agents targeting TrxR. SecTRAPs are prooxidant killers of cells, triggering mechanisms beyond those of a mere loss of thioredoxin reductase activity. PMID- 18382652 TI - Distinguishing molecular features and clinical characteristics of a putative new rhinovirus species, human rhinovirus C (HRV C). AB - BACKGROUND: Human rhinoviruses (HRVs) are the most frequently detected pathogens in acute respiratory tract infections (ARTIs) and yet little is known about the prevalence, recurrence, structure and clinical impact of individual members. During 2007, the complete coding sequences of six previously unknown and highly divergent HRV strains were reported. To catalogue the molecular and clinical features distinguishing the divergent HRV strains, we undertook, for the first time, in silico analyses of all available polyprotein sequences and performed retrospective reviews of the medical records of cases in which variants of the prototype strain, HRV-QPM, had been detected. METHODOLOGY/PRINCIPLE FINDINGS: Genomic analyses revealed that the six divergent strains, residing within a clade we previously called HRV A2, had the shortest polyprotein of all picornaviruses investigated. Structure-based amino acid alignments identified conserved motifs shared among members of the genus Rhinovirus as well as substantive deletions and insertions unique to the divergent strains. Deletions mostly affected regions encoding proteins traditionally involved in antigenicity and serving as HRV and HEV receptor footprints. Because the HRV A2 strains cannot yet be cultured, we created homology models of predicted HRV-QPM structural proteins. In silico comparisons confirmed that HRV-QPM was most closely related to the major group HRVs. HRV-QPM was most frequently detected in infants with expiratory wheezing or persistent cough who had been admitted to hospital and required supplemental oxygen. It was the only virus detected in 65% of positive individuals. These observations contributed to an objective clinical impact ranging from mild to severe. CONCLUSIONS: The divergent strains did not meet classification requirements for any existing species of the genus Rhinovirus or Enterovirus. HRV A2 strains should be partitioned into at least one new species, putatively called Human rhinovirus C, populated by members detected with high frequency, from individuals with respiratory symptoms requiring hospital admission. PMID- 18382653 TI - The airborne metagenome in an indoor urban environment. AB - The indoor atmosphere is an ecological unit that impacts on public health. To investigate the composition of organisms in this space, we applied culture independent approaches to microbes harvested from the air of two densely populated urban buildings, from which we analyzed 80 megabases genomic DNA sequence and 6000 16S rDNA clones. The air microbiota is primarily bacteria, including potential opportunistic pathogens commonly isolated from human inhabited environments such as hospitals, but none of the data contain matches to virulent pathogens or bioterror agents. Comparison of air samples with each other and nearby environments suggested that the indoor air microbes are not random transients from surrounding outdoor environments, but rather originate from indoor niches. Sequence annotation by gene function revealed specific adaptive capabilities enriched in the air environment, including genes potentially involved in resistance to desiccation and oxidative damage. This baseline index of air microbiota will be valuable for improving designs of surveillance for natural or man-made release of virulent pathogens. PMID- 18382654 TI - Inevitable evolutionary temporal elements in neural processing: a study based on evolutionary simulations. AB - Recent studies have suggested that some neural computational mechanisms are based on the fine temporal structure of spiking activity. However, less effort has been devoted to investigating the evolutionary aspects of such mechanisms. In this paper we explore the issue of temporal neural computation from an evolutionary point of view, using a genetic simulation of the evolutionary development of neural systems. We evolve neural systems in an environment with selective pressure based on mate finding, and examine the temporal aspects of the evolved systems. In repeating evolutionary sessions, there was a significant increase during evolution in the mutual information between the evolved agent's temporal neural representation and the external environment. In ten different simulated evolutionary sessions, there was an increased effect of time-related neural ablations on the agents' fitness. These results suggest that in some fitness landscapes the emergence of temporal elements in neural computation is almost inevitable. Future research using similar evolutionary simulations may shed new light on various biological mechanisms. PMID- 18382655 TI - Maternal TLR4 and NOD2 gene variants, pro-inflammatory phenotype and susceptibility to early-onset preeclampsia and HELLP syndrome. AB - BACKGROUND: Altered maternal inflammatory responses play a role in the development of preeclampsia and the hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome. We examined whether allelic variants of the innate immune receptors Toll-like receptor 4 (TLR4) and nucleotide-binding oligomerization domain 2 (NOD2), that impair the inflammatory response to endotoxin, are related to preeclampsia and HELLP syndrome. METHODS AND FINDINGS: We determined five common mutations in TLR4 (D299G and T399I) and NOD2 (R702W, G908R and L1007fs) in 340 primiparous women with a history of early-onset preeclampsia, of whom 177 women developed HELLP syndrome and in 113 women with a history of only uneventful pregnancies as controls. In addition, we assessed plasma levels of pro-inflammatory biomarkers C-reactive protein, interleukin-6, soluble intercellular adhesion molecule-1, fibrinogen and von Willebrand factor in a subset of 214 women included at least six months after delivery. After adjustment for maternal age and chronic hypertension, attenuating allelic variants of TLR4 were more common in women with a history of early-onset preeclampsia than in controls (OR 2.9 [95% CI 1.2-6.7]). Highest frequencies for TLR4 variants were observed in women who developed HELLP syndrome (adjusted OR 4.1 [95% CI 1.7-9.8]). In addition, high levels of interleukin-6 and fibrinogen were associated with a history of early-onset preeclampsia. Combined positivity for any of the TLR4 and NOD2 allelic variants and high levels of interleukin-6 was 6.9-fold more common in women with a history of early-onset preeclampsia (95% CI 2.1-23.2) compared to controls. CONCLUSIONS: We observed an association of common TLR4 and NOD2 gene variants, and pro-inflammatory phenotype with a history of early-onset preeclampsia and HELLP syndrome. These findings suggest involvement of the maternal innate immune system in severe hypertensive disorders of pregnancy. PMID- 18382656 TI - The CD81 partner EWI-2wint inhibits hepatitis C virus entry. AB - Two to three percent of the world's population is chronically infected with hepatitis C virus (HCV) and thus at risk of developing liver cancer. Although precise mechanisms regulating HCV entry into hepatic cells are still unknown, several cell surface proteins have been identified as entry factors for this virus. Among these molecules, the tetraspanin CD81 is essential for HCV entry. Here, we have identified a partner of CD81, EWI-2wint, which is expressed in several cell lines but not in hepatocytes. Ectopic expression of EWI-2wint in a hepatoma cell line susceptible to HCV infection blocked viral entry by inhibiting the interaction between the HCV envelope glycoproteins and CD81. This finding suggests that, in addition to the presence of specific entry factors in the hepatocytes, the lack of a specific inhibitor can contribute to the hepatotropism of HCV. This is the first example of a pathogen gaining entry into host cells that lack a specific inhibitory factor. PMID- 18382657 TI - Formation of toxic oligomeric alpha-synuclein species in living cells. AB - BACKGROUND: Misfolding, oligomerization, and fibrillization of alpha-synuclein are thought to be central events in the onset and progression of Parkinson's disease (PD) and related disorders. Although fibrillar alpha-synuclein is a major component of Lewy bodies (LBs), recent data implicate prefibrillar, oligomeric intermediates as the toxic species. However, to date, oligomeric species have not been identified in living cells. METHODOLOGY/PRINCIPAL FINDINGS: Here we used bimolecular fluorescence complementation (BiFC) to directly visualize alpha synuclein oligomerization in living cells, allowing us to study the initial events leading to alpha-synuclein oligomerization, the precursor to aggregate formation. This novel assay provides us with a tool with which to investigate how manipulations affecting alpha-synuclein aggregation affect the process over time. Stabilization of alpha-synuclein oligomers via BiFC results in increased cytotoxicity, which can be rescued by Hsp70 in a process that reduces the formation of alpha-synuclein oligomers. Introduction of PD-associated mutations in alpha-synuclein did not affect oligomer formation but the biochemical properties of the mutant alpha-synuclein oligomers differ from those of wild type alpha-synuclein. CONCLUSIONS/SIGNIFICANCE: This novel application of the BiFC assay to the study of the molecular basis of neurodegenerative disorders enabled the direct visualization of alpha-synuclein oligomeric species in living cells and its modulation by Hsp70, constituting a novel important tool in the search for therapeutics for synucleinopathies. PMID- 18382658 TI - FAK/src-family dependent activation of the Ste20-like kinase SLK is required for microtubule-dependent focal adhesion turnover and cell migration. AB - Cell migration involves a multitude of signals that converge on cytoskeletal reorganization, essential for development, immune responses and tissue repair. Using knockdown and dominant negative approaches, we show that the microtubule associated Ste20-like kinase SLK is required for focal adhesion turnover and cell migration downstream of the FAK/c-src complex. Our results show that SLK co localizes with paxillin, Rac1 and the microtubules at the leading edge of migrating cells and is activated by scratch wounding. SLK activation is dependent on FAK/c-src/MAPK signaling, whereas SLK recruitment to the leading edge is src dependent but FAK independent. Our results show that SLK represents a novel focal adhesion disassembly signal. PMID- 18382659 TI - Activin in the brain modulates anxiety-related behavior and adult neurogenesis. AB - Activin, a member of the transforming growth factor-beta superfamily, is an endocrine hormone that regulates differentiation and proliferation of a wide variety of cells. In the brain, activin protects neurons from ischemic damage. In this study, we demonstrate that activin modulates anxiety-related behavior by analyzing ACM4 and FSM transgenic mice in which activin and follistatin (which antagonizes the activin signal), respectively, were overexpressed in a forebrain specific manner under the control of the alphaCaMKII promoter. Behavioral analyses revealed that FSM mice exhibited enhanced anxiety compared to wild-type littermates, while ACM4 mice showed reduced anxiety. Importantly, survival of newly formed neurons in the subgranular zone of adult hippocampus was significantly decreased in FSM mice, which was partially rescued in ACM4/FSM double transgenic mice. Our findings demonstrate that the level of activin in the adult brain bi-directionally influences anxiety-related behavior. These results further suggest that decreases in postnatal neurogenesis caused by activin inhibition affect an anxiety-related behavior in adulthood. Activin and its signaling pathway may represent novel therapeutic targets for anxiety disorder as well as ischemic brain injury. PMID- 18382660 TI - Glucokinase gene mutations: structural and genotype-phenotype analyses in MODY children from South Italy. AB - BACKGROUND: Maturity onset diabetes of the young type 2 (or GCK MODY) is a genetic form of diabetes mellitus provoked by mutations in the glucokinase gene (GCK). METHODOLOGY/PRINCIPAL FINDINGS: We screened the GCK gene by direct sequencing in 30 patients from South Italy with suspected MODY. The mutation induced structural alterations in the protein were analyzed by molecular modeling. The patients' biochemical, clinical and anamnestic data were obtained. Mutations were detected in 16/30 patients (53%); 9 of the 12 mutations identified were novel (p.Glu70Asp, p.Phe123Leu, p.Asp132Asn, p.His137Asp, p.Gly162Asp, p.Thr168Ala, p.Arg392Ser, p.Glu290X, p.Gln106_Met107delinsLeu) and are in regions involved in structural rearrangements required for catalysis. The prevalence of mutation sites was higher in the small domain (7/12: approximately 59%) than in the large (4/12: 33%) domain or in the connection (1/12: 8%) region of the protein. Mild diabetic phenotypes were detected in almost all patients [mean (SD) OGTT = 7.8 mMol/L (1.8)] and mean triglyceride levels were lower in mutated than in unmutated GCK patients (p = 0.04). CONCLUSIONS: The prevalence of GCK MODY is high in southern Italy, and the GCK small domain is a hot spot for MODY mutations. Both the severity of the GCK mutation and the genetic background seem to play a relevant role in the GCK MODY phenotype. Indeed, a partial genotype phenotype correlation was identified in related patients (3 pairs of siblings) but not in two unrelated children bearing the same mutation. Thus, the molecular approach allows the physician to confirm the diagnosis and to predict severity of the mutation. PMID- 18382661 TI - Pharmacokinetic genes do not influence response or tolerance to citalopram in the STAR*D sample. AB - BACKGROUND: We sought to determine whether clinical response or tolerance to the Selective Serotonin Reuptake Inhibitor (SSRI) citalopram is associated with genetic polymorphisms in potentially relevant pharmacokinetic enzymes. METHODOLOGY: We used a two-stage case-control study design in which we split the sample of 1,953 subjects from the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) trial into a discovery (n = 831) and validation set (n = 1,046). Fifteen polymorphisms from five (CYP2D6, ABCB1, CYP2C19, CYP3A4, and CYP3A5) pharmacokinetic genes were genotyped. We examined the associations between these polymorphisms and citalopram response and tolerance. Significant associations were validated in the second stage for those polymorphism found to be statistically significant in the first stage. CONCLUSIONS: No genetic polymorphism in the pharmacokinetic genes examined was significantly associated with our response or tolerance phenotypes in both stages. For managing pharmacological treatment with citalopram, routine screening of the common pharmacokinetic DNA variants that we examined appears to be of limited clinical utility. PMID- 18382662 TI - A cryptic frizzled module in cell surface collagen 18 inhibits Wnt/beta-catenin signaling. AB - Collagens contain cryptic polypeptide modules that regulate major cell functions, such as cell proliferation or death. Collagen XVIII (C18) exists as three amino terminal end variants with specific amino terminal polypeptide modules. We investigated the function of the variant 3 of C18 (V3C18) containing a frizzled module (FZC18), which carries structural identity with the extracellular cysteine rich domain of the frizzled receptors. We show that V3C18 is a cell surface heparan sulfate proteoglycan, its topology being mediated by the FZC18 module. V3C18 mRNA was expressed at low levels in 21 normal adult human tissues. Its expression was up-regulated in fibrogenesis and in small well-differentiated liver tumors, but decreased in advanced human liver cancers. Low FZC18 immunostaining in liver cancer nodules correlated with markers of high Wnt/beta catenin activity. V3C18 (M(r) = 170 kD) was proteolytically processed into a cell surface FZC18-containing 50 kD glycoprotein precursor that bound Wnt3a in vitro through FZC18 and suppressed Wnt3a-induced stabilization of beta-catenin. Ectopic expression of either FZC18 (35 kD) or its 50 kD precursor inhibited Wnt/beta catenin signaling in colorectal and liver cancer cell lines, thus downregulating major cell cycle checkpoint gatekeepers cyclin D1 and c-myc and reducing tumor cell growth. By contrast, full-length V3C18 was unable to inhibit Wnt signaling. In summary, we identified a cell-surface signaling pathway whereby FZC18 inhibits Wnt/beta-catenin signaling. The signal, encrypted within cell-surface C18, is released by enzymatic processing as an active frizzled cysteine-rich domain (CRD) that reduces cancer cell growth. Thus, extracellular matrix controls Wnt signaling through a collagen-embedded CRD behaving as a cell-surface sensor of proteolysis, conveying feedback cues to control cancer cell fate. PMID- 18382663 TI - Anesthetics impact the resolution of inflammation. AB - BACKGROUND: Local and volatile anesthetics are widely used for surgery. It is not known whether anesthetics impinge on the orchestrated events in spontaneous resolution of acute inflammation. Here we investigated whether a commonly used local anesthetic (lidocaine) and a widely used inhaled anesthetic (isoflurane) impact the active process of resolution of inflammation. METHODS AND FINDINGS: Using murine peritonitis induced by zymosan and a systems approach, we report that lidocaine delayed and blocked key events in resolution of inflammation. Lidocaine inhibited both PMN apoptosis and macrophage uptake of apoptotic PMN, events that contributed to impaired PMN removal from exudates and thereby delayed the onset of resolution of acute inflammation and return to homeostasis. Lidocaine did not alter the levels of specific lipid mediators, including pro inflammatory leukotriene B(4), prostaglandin E(2) and anti-inflammatory lipoxin A(4), in the cell-free peritoneal lavages. Addition of a lipoxin A(4) stable analog, partially rescued lidocaine-delayed resolution of inflammation. To identify protein components underlying lidocaine's actions in resolution, systematic proteomics was carried out using nanospray-liquid chromatography tandem mass spectrometry. Lidocaine selectively up-regulated pro-inflammatory proteins including S100A8/9 and CRAMP/LL-37, and down-regulated anti-inflammatory and some pro-resolution peptides and proteins including IL-4, IL-13, TGF-a and Galectin-1. In contrast, the volatile anesthetic isoflurane promoted resolution in this system, diminishing the amplitude of PMN infiltration and shortening the resolution interval (Ri) approximately 50%. In addition, isoflurane down regulated a panel of pro-inflammatory chemokines and cytokines, as well as proteins known to be active in cell migration and chemotaxis (i.e., CRAMP and cofilin-1). The distinct impact of lidocaine and isoflurane on selective molecules may underlie their opposite actions in resolution of inflammation, namely lidocaine delayed the onset of resolution (T(max)), while isoflurane shortened resolution interval (Ri). CONCLUSIONS: Taken together, both local and volatile anesthetics impact endogenous resolution program(s), altering specific resolution indices and selective cellular/molecular components in inflammation resolution. Isoflurane enhances whereas lidocaine impairs timely resolution of acute inflammation. PMID- 18382664 TI - Physiology, pathology and relatedness of human tissues from gene expression meta analysis. AB - BACKGROUND: Development and maintenance of the identity of tissues is of central importance for multicellular organisms. Based on gene expression profiles, it is possible to divide genes in housekeeping genes and those whose expression is preferential in one or a few tissues and which provide specialized functions that have a strong effect on the physiology of the whole organism. RESULTS: We have surveyed the gene expression in 78 normal human tissues integrating publicly available microarray gene expression data. A total amount of 1601 genes were identified as selectively expressed in one or more tissues. The tissue-selective genes covered a wide range of cellular and molecular functions, and could be linked to 361 human diseases with Mendelian inheritance. Based on the gene expression profiles, we were able to form a network of tissues reflecting their functional relatedness and, to certain extent, their development. Using co citation driven gene network technique and promoter analysis, we predicted a transcriptional module where the co-operation of the transcription factors E2F and NF-kappaB can possibly regulate a number of genes involved in the neurogenesis that takes place in the adult hippocampus. CONCLUSIONS: Here we propose that integration of gene expression data from Affymetrix GeneChip experiments is possible through re-annotation and commonly used pre-processing methods. We suggest that some functional aspects of the tissues can be explained by the co-operation of multiple transcription factors that regulate the expression of selected groups of genes. PMID- 18382665 TI - Ethnic related selection for an ADH Class I variant within East Asia. AB - BACKGROUND: The alcohol dehydrogenases (ADH) are widely studied enzymes and the evolution of the mammalian gene cluster encoding these enzymes is also well studied. Previous studies have shown that the ADH1B*47His allele at one of the seven genes in humans is associated with a decrease in the risk of alcoholism and the core molecular region with this allele has been selected for in some East Asian populations. As the frequency of ADH1B*47His is highest in East Asia, and very low in most of the rest of the world, we have undertaken more detailed investigation in this geographic region. METHODOLOGY/PRINCIPAL FINDINGS: Here we report new data on 30 SNPs in the ADH7 and Class I ADH region in samples of 24 populations from China and Laos. These populations cover a wide geographic region and diverse ethnicities. Combined with our previously published East Asian data for these SNPs in 8 populations, we have typed populations from all of the 6 major linguistic phyla (Altaic including Korean-Japanese and inland Altaic, Sino Tibetan, Hmong-Mien, Austro-Asiatic, Daic, and Austronesian). The ADH1B genotyping data are strongly related to ethnicity. Only some eastern ethnic phyla or subphyla (Korean-Japanese, Han Chinese, Hmong-Mien, Daic, and Austronesian) have a high frequency of ADH1B*47His. ADH1B haplotype data clustered the populations into linguistic subphyla, and divided the subphyla into eastern and western parts. In the Hmong-Mien and Altaic populations, the extended haplotype homozygosity (EHH) and relative EHH (REHH) tests for the ADH1B core were consistent with selection for the haplotype with derived SNP alleles. In the other ethnic phyla, the core showed only a weak signal of selection at best. CONCLUSIONS/SIGNIFICANCE: The selection distribution is more significantly correlated with the frequency of the derived ADH1B regulatory region polymorphism than the derived amino-acid altering allele ADH1B*47His. Thus, the real focus of selection may be the regulatory region. The obvious ethnicity-related distributions of ADH1B diversities suggest the existence of some culture-related selective forces that have acted on the ADH1B region. PMID- 18382666 TI - Efficient in vivo electroporation of the postnatal rodent forebrain. AB - Functional gene analysis in vivo represents still a major challenge in biomedical research. Here we present a new method for the efficient introduction of nucleic acids into the postnatal mouse forebrain. We show that intraventricular injection of DNA followed by electroporation induces strong expression of transgenes in radial glia, neuronal precursors and neurons of the olfactory system. We present two proof-of-principle experiments to validate our approach. First, we show that expression of a human isoform of the neural cell adhesion molecule (hNCAM-140) in radial glia cells induces their differentiation into cells showing a neural precursor phenotype. Second, we demonstrate that p21 acts as a cell cycle inhibitor for postnatal neural stem cells. This approach will represent an important tool for future studies of postnatal neurogenesis and of neural development in general. PMID- 18382667 TI - DJ-1 modulates alpha-synuclein aggregation state in a cellular model of oxidative stress: relevance for Parkinson's disease and involvement of HSP70. AB - BACKGROUND: Parkinson's disease (PD) is a neurodegenerative pathology whose molecular etiopathogenesis is not known. Novel contributions have come from familial forms of PD caused by alterations in genes with apparently unrelated physiological functions. The gene coding for alpha-synuclein (alpha-syn) (PARK1) has been investigated as alpha-syn is located in Lewy bodies (LB), intraneuronal inclusions in the substantia nigra (SN) of PD patients. A-syn has neuroprotective chaperone-like and antioxidant functions and is involved in dopamine storage and release. DJ-1 (PARK7), another family-PD-linked gene causing an autosomal recessive form of the pathology, shows antioxidant and chaperone-like activities too. METHODOLOGY/PRINCIPAL FINDINGS: The present study addressed the question whether alpha-syn and DJ-1 interact functionally, with a view to finding some mechanism linking DJ-1 inactivation and alpha-syn aggregation and toxicity. We developed an in vitro model of alpha-syn toxicity in the human neuroblastoma cell line SK-N-BE, influencing DJ-1 and alpha-syn intracellular concentrations by exogenous addition of the fusion proteins TAT-alpha-syn and TAT-DJ-1; DJ-1 was inactivated by the siRNA method. On a micromolar scale TAT-alpha-syn aggregated and triggered neurotoxicity, while on the nanomolar scale it was neuroprotective against oxidative stress (induced by H(2)O(2) or 6-hydroxydopamine). TAT-DJ-1 increased the expression of HSP70, while DJ-1 silencing made SK-N-BE cells more susceptible to oxidative challenge, rendering TAT-alpha-syn neurotoxic at nanomolar scale, with the appearance of TAT-alpha-syn aggregates. CONCLUSION/SIGNIFICANCE: DJ-1 inactivation may thus promote alpha-syn aggregation and the related toxicity, and in this model HSP70 is involved in the antioxidant response and in the regulation of alpha-syn fibril formation. PMID- 18382668 TI - Low vs. high radioiodine activity to ablate the thyroid after thyroidectomy for cancer: a randomized study. AB - BACKGROUND: Radioactive iodine is commonly administered following thyroidectomy for differentiated thyroid carcinoma to ablate the thyroid remnant. The optimal administered activity of radioiodine is unknown. METHODOLOGY/PRINCIPAL FINDINGS: Adult subjects (n = 160) diagnosed with papillary or follicular thyroid carcinoma were randomly allocated to receive either 1100 MBq (30 mCi) or 3700 MBq (100 mCi) activity of radioiodine ((131)I) following thyroidectomy. The study participants were prepared for ablation using thyroid hormone withdrawal. Ablation was considered successful when serum thyroglobulin concentration was less than 1 ng/mL and no uptake was present in (131)I scan. Ablation was successful following one administration of radioiodine in 42 (52%; 95% CI, 41% to 63%) of the 81 evaluable study participants who received 1100 MBq, and in 43 (56%, 45% to 67%) of the 77 subjects who received 3700 MBq activity (P = .61). There was no difference between the groups in the numbers of repeat radioiodine treatments needed to complete ablation (P = .27). The higher activity was associated with more nausea and taste disturbances, and a longer stay in a radioprotected isolation unit. None of the participants died from thyroid cancer during a median follow up of 51 months; three subjects in the 3700 MBq group and none in the 1100 MBq group were diagnosed with distant metastases during follow-up. In a meta analysis of four randomized studies that compared the 1100 and 3700 MBq activities, the 1100 MBq activity tended to be associated with a higher risk of unsuccessful ablation (relative risk 1.148, 95% CI 0.974 to 1.353, P = .10). CONCLUSIONS/SIGNIFICANCE: The results provide no conclusive evidence that 3700 MBq activity is more effective for ablation of the thyroid remnant than 1100 MBq activity. The 3700 MBq activity is associated with more adverse effects. TRIAL REGISTRATION: ClinicalTrials.gov NCT00115895. PMID- 18382669 TI - Paracrine factors of mesenchymal stem cells recruit macrophages and endothelial lineage cells and enhance wound healing. AB - Bone marrow derived mesenchymal stem cells (BM-MSCs) have been shown to enhance wound healing; however, the mechanisms involved are barely understood. In this study, we examined paracrine factors released by BM-MSCs and their effects on the cells participating in wound healing compared to those released by dermal fibroblasts. Analyses of BM-MSCs with Real-Time PCR and of BM-MSC-conditioned medium by antibody-based protein array and ELISA indicated that BM-MSCs secreted distinctively different cytokines and chemokines, such as greater amounts of VEGF alpha, IGF-1, EGF, keratinocyte growth factor, angiopoietin-1, stromal derived factor-1, macrophage inflammatory protein-1alpha and beta and erythropoietin, compared to dermal fibroblasts. These molecules are known to be important in normal wound healing. BM-MSC-conditioned medium significantly enhanced migration of macrophages, keratinocytes and endothelial cells and proliferation of keratinocytes and endothelial cells compared to fibroblast-conditioned medium. Moreover, in a mouse model of excisional wound healing, where concentrated BM-MSC conditioned medium was applied, accelerated wound healing occurred compared to administration of pre-conditioned or fibroblast-conditioned medium. Analysis of cell suspensions derived from the wound by FACS showed that wounds treated with BM-MSC-conditioned medium had increased proportions of CD4/80-positive macrophages and Flk-1-, CD34- or c-kit-positive endothelial (progenitor) cells compared to wounds treated with pre-conditioned medium or fibroblast-conditioned medium. Consistent with the above findings, immunohistochemical analysis of wound sections showed that wounds treated with BM-MSC-conditioned medium had increased abundance of macrophages. Our results suggest that factors released by BM-MSCs recruit macrophages and endothelial lineage cells into the wound thus enhancing wound healing. PMID- 18382670 TI - Different effect of proteasome inhibition on vesicular stomatitis virus and poliovirus replication. AB - Proteasome activity is an important part of viral replication. In this study, we examined the effect of proteasome inhibitors on the replication of vesicular stomatitis virus (VSV) and poliovirus. We found that the proteasome inhibitors significantly suppressed VSV protein synthesis, virus accumulation, and protected infected cells from toxic effect of VSV replication. In contrast, poliovirus replication was delayed, but not diminished in the presence of the proteasome inhibitors MG132 and Bortezomib. We also found that inhibition of proteasomes stimulated stress-related processes, such as accumulation of chaperone hsp70, phosphorylation of eIF2alpha, and overall inhibition of translation. VSV replication was sensitive to this stress with significant decline in replication process. Poliovirus growth was less sensitive with only delay in replication. Inhibition of proteasome activity suppressed cellular and VSV protein synthesis, but did not reduce poliovirus protein synthesis. Protein kinase GCN2 supported the ability of proteasome inhibitors to attenuate general translation and to suppress VSV replication. We propose that different mechanisms of translational initiation by VSV and poliovirus determine their sensitivity to stress induced by the inhibition of proteasomes. To our knowledge, this is the first study that connects the effect of stress induced by proteasome inhibition with the efficiency of viral infection. PMID- 18382671 TI - Three-tiered risk stratification model to predict progression in Barrett's esophagus using epigenetic and clinical features. AB - BACKGROUND: Barrett's esophagus predisposes to esophageal adenocarcinoma. However, the value of endoscopic surveillance in Barrett's esophagus has been debated because of the low incidence of esophageal adenocarcinoma in Barrett's esophagus. Moreover, high inter-observer and sampling-dependent variation in the histologic staging of dysplasia make clinical risk assessment problematic. In this study, we developed a 3-tiered risk stratification strategy, based on systematically selected epigenetic and clinical parameters, to improve Barrett's esophagus surveillance efficiency. METHODS AND FINDINGS: We defined high-grade dysplasia as endpoint of progression, and Barrett's esophagus progressor patients as Barrett's esophagus patients with either no dysplasia or low-grade dysplasia who later developed high-grade dysplasia or esophageal adenocarcinoma. We analyzed 4 epigenetic and 3 clinical parameters in 118 Barrett's esophagus tissues obtained from 35 progressor and 27 non-progressor Barrett's esophagus patients from Baltimore Veterans Affairs Maryland Health Care Systems and Mayo Clinic. Based on 2-year and 4-year prediction models using linear discriminant analysis (area under the receiver-operator characteristic (ROC) curve: 0.8386 and 0.7910, respectively), Barrett's esophagus specimens were stratified into high risk (HR), intermediate-risk (IR), or low-risk (LR) groups. This 3-tiered stratification method retained both the high specificity of the 2-year model and the high sensitivity of the 4-year model. Progression-free survivals differed significantly among the 3 risk groups, with p = 0.0022 (HR vs. IR) and p<0.0001 (HR or IR vs. LR). Incremental value analyses demonstrated that the number of methylated genes contributed most influentially to prediction accuracy. CONCLUSIONS: This 3-tiered risk stratification strategy has the potential to exert a profound impact on Barrett's esophagus surveillance accuracy and efficiency. PMID- 18382672 TI - Biofilm induced tolerance towards antimicrobial peptides. AB - Increased tolerance to antimicrobial agents is thought to be an important feature of microbes growing in biofilms. We address the question of how biofilm organization affects antibiotic susceptibility. We established Escherichia coli biofilms with differential structural organization due to the presence of IncF plasmids expressing altered forms of the transfer pili in two different biofilm model systems. The mature biofilms were subsequently treated with two antibiotics with different molecular targets, the peptide antibiotic colistin and the fluoroquinolone ciprofloxacin. The dynamics of microbial killing were monitored by viable count determination, and confocal laser microscopy. Strains forming structurally organized biofilms show an increased bacterial survival when challenged with colistin, compared to strains forming unstructured biofilms. The increased survival is due to genetically regulated tolerant subpopulation formation and not caused by a general biofilm property. No significant difference in survival was detected when the strains were challenged with ciprofloxacin. Our data show that biofilm formation confers increased colistin tolerance to cells within the biofilm structure, but the protection is conditional being dependent on the structural organization of the biofilm, and the induction of specific tolerance mechanisms. PMID- 18382673 TI - Mesoscopic structure conditions the emergence of cooperation on social networks. AB - BACKGROUND: We study the evolutionary Prisoner's Dilemma on two social networks substrates obtained from actual relational data. METHODOLOGY/PRINCIPAL FINDINGS: We find very different cooperation levels on each of them that cannot be easily understood in terms of global statistical properties of both networks. We claim that the result can be understood at the mesoscopic scale, by studying the community structure of the networks. We explain the dependence of the cooperation level on the temptation parameter in terms of the internal structure of the communities and their interconnections. We then test our results on community structured, specifically designed artificial networks, finding a good agreement with the observations in both real substrates. CONCLUSION: Our results support the conclusion that studies of evolutionary games on model networks and their interpretation in terms of global properties may not be sufficient to study specific, real social systems. Further, the study allows us to define new quantitative parameters that summarize the mesoscopic structure of any network. In addition, the community perspective may be helpful to interpret the origin and behavior of existing networks as well as to design structures that show resilient cooperative behavior. PMID- 18382674 TI - Ultrasonic vocalizations induced by sex and amphetamine in M2, M4, M5 muscarinic and D2 dopamine receptor knockout mice. AB - Adult mice communicate by emitting ultrasonic vocalizations (USVs) during the appetitive phases of sexual behavior. However, little is known about the genes important in controlling call production. Here, we study the induction and regulation of USVs in muscarinic and dopaminergic receptor knockout (KO) mice as well as wild-type controls during sexual behavior. Female mouse urine, but not female rat or human urine, induced USVs in male mice, whereas male urine did not induce USVs in females. Direct contact of males with females is required for eliciting high level of USVs in males. USVs (25 to120 kHz) were emitted only by males, suggesting positive state; however human-audible squeaks were produced only by females, implying negative state during male-female pairing. USVs were divided into flat and frequency-modulated calls. Male USVs often changed from continuous to broken frequency-modulated calls after initiation of mounting. In M2 KO mice, USVs were lost in about 70-80% of the mice, correlating with a loss of sexual interaction. In M5 KO mice, mean USVs were reduced by almost 80% even though sexual interaction was vigorous. In D2 KOs, the duration of USVs was extended by 20%. In M4 KOs, no significant differences were observed. Amphetamine dose-dependently induced USVs in wild-type males (most at 0.5 mg/kg i.p.), but did not elicit USVs in M5 KO or female mice. These studies suggest that M2 and M5 muscarinic receptors are needed for male USV production during male-female interactions, likely via their roles in dopamine activation. These findings are important for the understanding of the neural substrates for positive affect. PMID- 18382675 TI - Appetite enhancement and weight gain by peripheral administration of TrkB agonists in non-human primates. AB - Loss of function mutations in the receptor tyrosine kinase TrkB pathway resulted in hyperphagia and morbid obesity in human and rodents. Conversely, peripheral or central stimulation of TrkB by its natural ligands BDNF or NT4 reduced body weight and food intake in mice, supporting the idea that TrkB is a key anorexigenic signal downstream of the melanocortin-4 receptor (Mc4r) system. Here we show that in non-human primates TrkB agonists were anorexigenic when applied centrally, but surprisingly orexigenic, leading to gain in appetite, body weight, fat deposits and serum leptin levels, when given peripherally. The orexigenic and pro-obesity effects of peripherally administered TrkB agonists appear to be dose dependent, not associated with fluid retention nor with evidence of receptor down regulation. Our findings revealed that TrkB signaling exerts dual control on energy homeostasis in the primates that could be targeted for the treatment of either wasting disorders or obesity. PMID- 18382676 TI - Quantitative multicolor compositional imaging resolves molecular domains in cell matrix adhesions. AB - BACKGROUND: Cellular processes occur within dynamic and multi-molecular compartments whose characterization requires analysis at high spatio-temporal resolution. Notable examples for such complexes are cell-matrix adhesion sites, consisting of numerous cytoskeletal and signaling proteins. These adhesions are highly variable in their morphology, dynamics, and apparent function, yet their molecular diversity is poorly defined. METHODOLOGY/PRINCIPAL FINDINGS: We present here a compositional imaging approach for the analysis and display of multi component compositions. This methodology is based on microscopy-acquired multicolor data, multi-dimensional clustering of pixels according to their composition similarity and display of the cellular distribution of these composition clusters. We apply this approach for resolving the molecular complexes associated with focal-adhesions, and the time-dependent effects of Rho kinase inhibition. We show here compositional variations between adhesion sites, as well as ordered variations along the axis of individual focal-adhesions. The multicolor clustering approach also reveals distinct sensitivities of different focal-adhesion-associated complexes to Rho-kinase inhibition. CONCLUSIONS/SIGNIFICANCE: Multicolor compositional imaging resolves "molecular signatures" characteristic to focal-adhesions and related structures, as well as sub-domains within these adhesion sites. This analysis enhances the spatial information with additional "contents-resolved" dimensions. We propose that compositional imaging can serve as a powerful tool for studying complex multi molecular assemblies in cells and for mapping their distribution at sub-micron resolution. PMID- 18382677 TI - The p12 domain is unstructured in a murine leukemia virus p12-CA(N) Gag construct. AB - The Gag polyproteins of gammaretroviruses contain a conserved p12 domain between MA and CA that plays critical roles in virus assembly, reverse transcription and nuclear integration. Here we show using nuclear magnetic resonance, that p12 is unstructured in a Moloney murine leukemia virus (MMLV) Gag fragment that includes the N-terminal domain of CA (p12-CA(N)). Furthermore, no long range interactions were observed between the domains, as has been previously predicted. Flexibility appears to be a common feature of Gag "late" domains required for virus release during budding. Residues near the N-terminus of CA(N) that form a beta-hairpin in the mature CA protein are unfolded in p12-CA(N), consistent with proposals that hairpin formation helps trigger capsid assembly. PMID- 18382678 TI - GPCR genes are preferentially retained after whole genome duplication. AB - One of the most interesting questions in biology is whether certain pathways have been favored during evolution, and if so, what properties could cause such a preference. Due to the lack of experimental evidence, whether select gene families have been preferentially retained over time after duplication in metazoan organisms remains unclear. Here, by syntenic mapping of nonchemosensory G protein-coupled receptor genes (nGPCRs which represent half the receptome for transmembrane signaling) in the vertebrate genomes, we found that, as opposed to the 8-15% retention rate for whole genome duplication (WGD)-derived gene duplicates in the entire genome of pufferfish, greater than 27.8% of WGD-derived nGPCRs which interact with a nonpeptide ligand were retained after WGD in pufferfish Tetraodon nigroviridis. In addition, we show that concurrent duplication of cognate ligand genes by WGD could impose selection of nGPCRs that interact with a polypeptide ligand. Against less than 2.25% probability for parallel retention of a pair of WGD-derived ligands and a pair of cognate receptor duplicates, we found a more than 8.9% retention of WGD-derived ligand nGPCR pairs--threefold greater than one would surmise. These results demonstrate that gene retention is not uniform after WGD in vertebrates, and suggest a Darwinian selection of GPCR-mediated intercellular communication in metazoan organisms. PMID- 18382679 TI - Distinct roles of jasmonates and aldehydes in plant-defense responses. AB - BACKGROUND: Many inducible plant-defense responses are activated by jasmonates (JAs), C(6)-aldehydes, and their corresponding derivatives, produced by the two main competing branches of the oxylipin pathway, the allene oxide synthase (AOS) and hydroperoxide lyase (HPL) branches, respectively. In addition to competition for substrates, these branch-pathway-derived metabolites have substantial overlap in regulation of gene expression. Past experiments to define the role of C(6) aldehydes in plant defense responses were biased towards the exogenous application of the synthetic metabolites or the use of genetic manipulation of HPL expression levels in plant genotypes with intact ability to produce the competing AOS-derived metabolites. To uncouple the roles of the C(6)-aldehydes and jasmonates in mediating direct and indirect plant-defense responses, we generated Arabidopsis genotypes lacking either one or both of these metabolites. These genotypes were subsequently challenged with a phloem-feeding insect (aphids: Myzus persicae), an insect herbivore (leafminers: Liriomyza trifolii), and two different necrotrophic fungal pathogens (Botrytis cinerea and Alternaria brassicicola). We also characterized the volatiles emitted by these plants upon aphid infestation or mechanical wounding and identified hexenyl acetate as the predominant compound in these volatile blends. Subsequently, we examined the signaling role of this compound in attracting the parasitoid wasp (Aphidius colemani), a natural enemy of aphids. PRINCIPAL FINDINGS: This study conclusively establishes that jasmonates and C(6)-aldehydes play distinct roles in plant defense responses. The jasmonates are indispensable metabolites in mediating the activation of direct plant-defense responses, whereas the C(6)-aldehyes are not. On the other hand, hexenyl acetate, an acetylated C(6)-aldehyde, is the predominant wound-inducible volatile signal that mediates indirect defense responses by directing tritrophic (plant-herbivore-natural enemy) interactions. SIGNIFICANCE: The data suggest that jasmonates and hexenyl acetate play distinct roles in mediating direct and indirect plant-defense responses. The potential advantage of this "division of labor" is to ensure the most effective defense strategy that minimizes incurred damages at a reduced metabolic cost. PMID- 18382680 TI - Ecological niche dimensionality and the evolutionary diversification of stick insects. AB - The degree of phenotypic divergence and reproductive isolation between taxon pairs can vary quantitatively, and often increases as evolutionary divergence proceeds through various stages, from polymorphism to population differentiation, ecotype and race formation, speciation, and post-speciational divergence. Although divergent natural selection promotes divergence, it does not always result in strong differentiation. For example, divergent selection can fail to complete speciation, and distinct species pairs sometimes collapse ('speciation in reverse'). Widely-discussed explanations for this variability concern genetic architecture, and the geographic arrangement of populations. A less-explored possibility is that the degree of phenotypic and reproductive divergence between taxon pairs is positively related to the number of ecological niche dimensions (i.e., traits) subject to divergent selection. Some data supporting this idea stem from laboratory experimental evolution studies using Drosophila, but tests from nature are lacking. Here we report results from manipulative field experiments in natural populations of herbivorous Timema stick insects that are consistent with this 'niche dimensionality' hypothesis. In such insects, divergent selection between host plants might occur for cryptic colouration (camouflage to evade visual predation), physiology (to detoxify plant chemicals), or both of these niche dimensions. We show that divergent selection on the single niche dimension of cryptic colouration can result in ecotype formation and intermediate levels of phenotypic and reproductive divergence between populations feeding on different hosts. However, greater divergence between a species pair involved divergent selection on both niche dimensions. Although further replication of the trends reported here is required, the results suggest that dimensionality of selection may complement genetic and geographic explanations for the degree of diversification in nature. PMID- 18382681 TI - An integrated approach to the prediction of chemotherapeutic response in patients with breast cancer. AB - BACKGROUND: A major challenge in oncology is the selection of the most effective chemotherapeutic agents for individual patients, while the administration of ineffective chemotherapy increases mortality and decreases quality of life in cancer patients. This emphasizes the need to evaluate every patient's probability of responding to each chemotherapeutic agent and limiting the agents used to those most likely to be effective. METHODS AND RESULTS: Using gene expression data on the NCI-60 and corresponding drug sensitivity, mRNA and microRNA profiles were developed representing sensitivity to individual chemotherapeutic agents. The mRNA signatures were tested in an independent cohort of 133 breast cancer patients treated with the TFAC (paclitaxel, 5-fluorouracil, adriamycin, and cyclophosphamide) chemotherapy regimen. To further dissect the biology of resistance, we applied signatures of oncogenic pathway activation and performed hierarchical clustering. We then used mRNA signatures of chemotherapy sensitivity to identify alternative therapeutics for patients resistant to TFAC. Profiles from mRNA and microRNA expression data represent distinct biologic mechanisms of resistance to common cytotoxic agents. The individual mRNA signatures were validated in an independent dataset of breast tumors (P = 0.002, NPV = 82%). When the accuracy of the signatures was analyzed based on molecular variables, the predictive ability was found to be greater in basal-like than non basal-like patients (P = 0.03 and P = 0.06). Samples from patients with co-activated Myc and E2F represented the cohort with the lowest percentage (8%) of responders. Using mRNA signatures of sensitivity to other cytotoxic agents, we predict that TFAC non-responders are more likely to be sensitive to docetaxel (P = 0.04), representing a viable alternative therapy. CONCLUSIONS: Our results suggest that the optimal strategy for chemotherapy sensitivity prediction integrates molecular variables such as ER and HER2 status with corresponding microRNA and mRNA expression profiles. Importantly, we also present evidence to support the concept that analysis of molecular variables can present a rational strategy to identifying alternative therapeutic opportunities. PMID- 18382682 TI - Measurable prediction for the single patient and the results of large double blind controlled randomized trials. AB - BACKGROUND: It has been shown that the clinical state of one patient can be represented by known measured variables of interest, each of which then form the element of a fuzzy set as point in the unit hypercube. We hypothesized that precise comparison of a single patient with the average patient of a large double blind controlled randomized study is possible using fuzzy theory. METHODS/PRINCIPLE FINDINGS: The sets as points unit hypercube geometry allows fuzzy subsethood to define in measures of fuzzy cardinality different conditions, similarity and comparison between fuzzy sets. A fuzzy measure of prediction is defined from fuzzy measures of similarity and comparison. It is a measure of the degree to which fuzzy set A is similar to fuzzy set B when different conditions are taken into account and removed from the comparison. When represented as a fuzzy set as point in the unit hypercube, a clinical patient can be compared to an average patient of a large group study in a precise manner. This comparison is expressed by the fuzzy prediction measure. This measure in itself is not a probability. Once thus precisely matched to the average patient of a large group study, risk reduction is calculated by multiplying the measured similarity of the clinical patient to the risk of the average trial patient. CONCLUSION/SIGNIFICANCE: Otherwise not precisely translatable to the single case, the result of group statistics can be applied to the single case through the use of fuzzy subsethood and measured in fuzzy cardinality. This measure is an alternative to a Bayesian or other probability based statistical approach. PMID- 18382683 TI - In vivo isolation and characterization of stem cells with diverse phenotypes using growth factor impregnated biomatrices. AB - BACKGROUND: The stimulation to differentiate into specific cell types for somatic stem cells is largely due to a series of internal and external signals coming from the microenvironment that surrounds the stem cell. Even though intensive research has been made, the basic mechanisms of plasticity and/or the molecules regulating stem cells proliferation and differentiation are not completely determined. Potential answers concerning the problems could be derived from the studies of stem cells in culture. METHODOLOGY/PRINCIPLE FINDINGS: We combine a new procedure (using the matrigel biopolymer supplemented with a selected cytokine/growth factor) with classic techniques such as light, confocal and electron microscopy, immunohistochemistry and cell culture, to perform an analysis on stem cells involved in the leech (Hirudo medicinalis) repair tissues. The leech has a relative anatomical simplicity and is a reliable model for studying a variety of basic events, such as tissue repair, which has a striking similarity with vertebrate responses. Our data demonstrate that the injection of an appropriate combination of the matrigel biopolymer supplemented with a selected cytokine/growth factor in the leech Hirudo medicinalis is a remarkably effective tool for isolating a specific cell population in vivo. A comparative analysis of biopolymer in vivo sorted stem cells indicates that VEGF recruited cells of a hematopoietic/endothelial phenotype whereas MCP-1/CCL2 isolated cells that were of an early myeloid lineage. CONCLUSION: Our paper describes, for the first time, a method allowing not only the isolation of a specific cell population in relation to the cytokine utilized but also the possibility to culture a precise cell type whose isolation is otherwise quite difficult. This approach could be broadly applied to isolate stem cells of diverse origins based on the recruitment stimuli employed. PMID- 18382684 TI - p27 deficiency cooperates with Bcl-2 but not Bax to promote T-cell lymphoma. AB - The effect of Bcl-2 on oncogenesis is complex and expression may either delay or accelerate oncogenesis. The pro-oncogenic activity is attributed to its well characterized anti-apoptotic function while the anti-oncogenic function has been attributed to its inhibition of cellular proliferation. Recent studies demonstrate that p27 may mediate the effects of Bcl-2 on cellular proliferation. We hypothesized that p27 may suppress tumor formation by Bcl-2 family members. To test this hypothesis, cell cycle inhibition and lymphoma development were examined in Lck-Bcl-2 and Lck-Bax38/1 transgenic mice deficient in p27. Strikingly, p27 deficiency synergistically cooperates with Bcl-2 to increase T cell hyperplasia and development of spontaneous T cell lymphomas. Within 1 year, >90% of these mice had developed thymic T cell lymphomas. This high penetrance contrasts with a one year incidence of <5% of thymic lymphoma in Lck-Bcl-2 or p27 -/- mice alone. In contrast, p27 deficiency had no effect on tumor formation in Lck-Bax38/1 transgenic mice, another model of T cell lymphoma. Histologically the lymphomas in p27 -/- Lck-Bcl-2 mice are lymphoblastic and frequently involve multiple organs suggesting an aggressive phenotype. Interestingly, in mature splenic T cells, Bcl-2 largely retains its anti-proliferative function even in the absence of p27. T cells from p27 -/- Lck-Bcl-2 mice show delayed kinetics of CDK2 Thr-160 phosphorylation. This delay is associated with a delay in the up regulation of both Cyclin D2 and D3. These data demonstrate a complex relationship between the Bcl-2 family, cellular proliferation, and oncogenesis and demonstrate that p27 up-regulation is not singularly important in the proliferative delay observed in T cells expressing Bcl-2 family members. Nonetheless, the results indicate that p27 is a critical tumor suppressor in the context of Bcl-2 expression. PMID- 18382685 TI - Gene expression signatures of radiation response are specific, durable and accurate in mice and humans. AB - BACKGROUND: Previous work has demonstrated the potential for peripheral blood (PB) gene expression profiling for the detection of disease or environmental exposures. METHODS AND FINDINGS: We have sought to determine the impact of several variables on the PB gene expression profile of an environmental exposure, ionizing radiation, and to determine the specificity of the PB signature of radiation versus other genotoxic stresses. Neither genotype differences nor the time of PB sampling caused any lessening of the accuracy of PB signatures to predict radiation exposure, but sex difference did influence the accuracy of the prediction of radiation exposure at the lowest level (50 cGy). A PB signature of sepsis was also generated and both the PB signature of radiation and the PB signature of sepsis were found to be 100% specific at distinguishing irradiated from septic animals. We also identified human PB signatures of radiation exposure and chemotherapy treatment which distinguished irradiated patients and chemotherapy-treated individuals within a heterogeneous population with accuracies of 90% and 81%, respectively. CONCLUSIONS: We conclude that PB gene expression profiles can be identified in mice and humans that are accurate in predicting medical conditions, are specific to each condition and remain highly accurate over time. PMID- 18382686 TI - Toll-like receptor ligands induce human T cell activation and death, a model for HIV pathogenesis. AB - BACKGROUND: Recently, heightened systemic translocation of microbial products was found in persons with chronic HIV infection and this was linked to immune activation and CD4(+) T cell homeostasis. METHODOLOGY: We examined here the effects of microbial Toll-like receptor (TLR) ligands on T cell activation in vitro. CONCLUSIONS/FINDINGS: We show that exposure to TLR ligands results in activation of memory and effector CD4(+) and CD8(+) T cells. After exposure to each of 8 different ligands that activate TLRs 2, 3, 4, 5, 7, 8, and 9, CD8(+) T cells are activated and gain expression of the C type lectin CD69 that may promote their retention in lymphoid tissues. In contrast, CD4(+) T cells rarely increase CD69 expression but instead enter cell cycle. Despite activation and cell cycle entry, CD4(+) T cells divide poorly and instead, disproportionately undergo activation-induced cell death. Systemic exposure to TLR agonists may therefore increase immune activation, effector cell sequestration in lymphoid tissues and T cell turnover. These events may contribute to the pathogenesis of immune dysfunction and CD4+ T cell losses in chronic infection with the human immunodeficiency virus. PMID- 18382687 TI - A comparison of donor-acceptor pairs for genetically encoded FRET sensors: application to the Epac cAMP sensor as an example. AB - We recently reported on CFP-Epac-YFP, an Epac-based single polypeptide FRET reporter to resolve cAMP levels in living cells. In this study, we compared and optimized the fluorescent protein donor/acceptor pairs for use in biosensors such as CFP-Epac-YFP. Our strategy was to prepare a wide range of constructs consisting of different donor and acceptor fluorescent proteins separated by a short linker. Constructs were expressed in HEK293 cells and tested for FRET and other relevant properties. The most promising pairs were subsequently used in an attempt to improve the FRET span of the Epac-based cAMP sensor. The results show significant albeit not perfect correlation between performance in the spacer construct and in the Epac sensor. Finally, this strategy enabled us to identify improved sensors both for detection by sensitized emission and by fluorescent lifetime imaging. The present overview should be helpful in guiding development of future FRET sensors. PMID- 18382688 TI - Maternal cocaine administration in mice alters DNA methylation and gene expression in hippocampal neurons of neonatal and prepubertal offspring. AB - Previous studies documented significant behavioral changes in the offspring of cocaine-exposed mothers. We now explore the hypothesis that maternal cocaine exposure could alter the fetal epigenetic machinery sufficiently to cause lasting neurochemical and functional changes in the offspring. Pregnant CD1 mice were administered either saline or 20 mg/kg cocaine twice daily on gestational days 8 19. Male pups from each of ten litters of the cocaine and control groups were analyzed at 3 (P3) or 30 (P30) days postnatum. Global DNA methylation, methylated DNA immunoprecipitation followed by CGI(2) microarray profiling and bisulfite sequencing, as well as quantitative real-time RT-PCR gene expression analysis, were evaluated in hippocampal pyramidal neurons excised by laser capture microdissection. Following maternal cocaine exposure, global DNA methylation was significantly decreased at P3 and increased at P30. Among the 492 CGIs whose methylation was significantly altered by cocaine at P3, 34% were hypermethylated while 66% were hypomethylated. Several of these CGIs contained promoter regions for genes implicated in crucial cellular functions. Endogenous expression of selected genes linked to the abnormally methylated CGIs was correspondingly decreased or increased by as much as 4-19-fold. By P30, some of the cocaine associated effects at P3 endured, reversed to opposite directions, or disappeared. Further, additional sets of abnormally methylated targets emerged at P30 that were not observed at P3. Taken together, these observations indicate that maternal cocaine exposure during the second and third trimesters of gestation could produce potentially profound structural and functional modifications in the epigenomic programs of neonatal and prepubertal mice. PMID- 18382690 TI - Regulation of the high affinity IgE receptor (Fc epsilonRI) in human neutrophils: role of seasonal allergen exposure and Th-2 cytokines. AB - The high affinity IgE receptor, Fc epsilonRI, plays a key role in the immunological pathways involved in allergic asthma. Previously we have demonstrated that human neutrophils isolated from allergic asthmatics express a functional Fc epsilonRI, and therefore it was of importance to examine the factors regulating its expression. In this study, we found that neutrophils from allergic asthmatics showed increased expression of Fc epsilonRI-alpha chain surface protein, total protein and mRNA compared with those from allergic non asthmatics and healthy donors (p<0.001). Interestingly, in neutrophils isolated from allergic asthmatics, Fc epsilonRI-alpha chain surface protein and mRNA expression were significantly greater during the pollen season than outside the pollen season (n = 9, P = 0.001), an effect which was not observed either in the allergic non asthmatic group or the healthy donors (p>0.05). Allergen exposure did not affect other surface markers of neutrophils such as CD16/Fc gammaRIII or IL-17R. In contrast to stimulation with IgE, neutrophils incubated with TH2 cytokines IL-9, GM-CSF, and IL-4, showed enhanced Fc epsilonRI-alpha chain surface expression. In conclusion, these results suggest that enhanced Fc epsilonRI expression in human neutrophils from allergic asthmatics during the pollen season can make them more susceptible to the biological effects of IgE, providing a possible new mechanism by which neutrophils contribute to allergic asthma. PMID- 18382689 TI - Association of mild anemia with cognitive, functional, mood and quality of life outcomes in the elderly: the "Health and Anemia" study. AB - BACKGROUND: In the elderly persons, hemoglobin concentrations slightly below the lower limit of normal are common, but scant evidence is available on their relationship with significant health indicators. The objective of the present study was to cross-sectionally investigate the association of mild grade anemia with cognitive, functional, mood, and quality of life (QoL) variables in community-dwelling elderly persons. METHODS: Among the 4,068 eligible individuals aged 65-84 years, all persons with mild anemia (n = 170) and a randomly selected sample of non-anemic controls (n = 547) were included in the study. Anemia was defined according to World Health Organization (WHO) criteria and mild grade anemia was defined as a hemoglobin concentration between 10.0 and 11.9 g/dL in women and between 10.0 and 12.9 g/dL in men. Cognition and functional status were assessed using measures of selective attention, episodic memory, cognitive flexibility and instrumental and basic activities of daily living. Mood and QoL were evaluated by means of the Geriatric Depression Scale-10, the Short-Form health survey (SF-12), and the Functional Assessment of Cancer Therapy-Anemia. RESULTS: In univariate analyses, mild anemic elderly persons had significantly worse results on almost all cognitive, functional, mood, and QoL measures. In multivariable logistic regressions, after adjustment for a large number of demographic and clinical confounders, mild anemia remained significantly associated with measures of selective attention and disease-specific QoL (all fully adjusted p<.046). When the lower limit of normal hemoglobin concentration according to WHO criteria was raised to define anemia (+0.2 g/dL), differences between mild anemic and non anemic elderly persons tended to increase on almost every variable. CONCLUSIONS: Cross-sectionally, mild grade anemia was independently associated with worse selective attention performance and disease specific QoL ratings. PMID- 18382691 TI - Erythropoietin: a potent inducer of peripheral immuno/inflammatory modulation in autoimmune EAE. AB - BACKGROUND: Beneficial effects of short-term erythropoietin (EPO) therapy have been demonstrated in several animal models of acute neurologic injury, including experimental autoimmune encephalomyelitis (EAE)--the animal model of multiple sclerosis. We have found that EPO treatment substantially reduces the acute clinical paralysis seen in EAE mice and this improvement is accompanied by a large reduction in the mononuclear cell infiltration and downregulation of glial MHC class II expression within the inflamed CNS. Other reports have recently indicated that peripherally generated anti-inflammatory CD4(+)Foxp3(+) regulatory T cells (Tregs) and the IL17-producing CD4+ T helper cell (Th17) subpopulations play key antagonistic roles in EAE pathogenesis. However, no information regarding the effects of EPO therapy on the behavior of the general mononuclear lymphocyte population, Tregs or Th17 cells in EAE has emerged. METHODS AND FINDINGS: We first determined in vivo that EPO therapy markedly suppressed MOG specific T cell proliferation and sharply reduced the number of reactive dendritic cells (CD11c positive) in EAE lymph nodes during both inductive and later symptomatic phases of MOG(35-55) induced EAE. We then determined the effect in vivo of EPO on numbers of peripheral Treg cells and Th17 cells. We found that EPO treatment modulated immune balance in both the periphery and the inflamed spinal cord by promoting a large expansion in Treg cells, inhibiting Th17 polarization and abrogating proliferation of the antigen presenting dendritic cell population. Finally we utilized tissue culture assays to show that exposure to EPO in vitro similarly downregulated MOG-specific T cell proliferation and also greatly suppressed T cell production of pro-inflammatory cytokines. CONCLUSIONS: Taken together, our findings reveal an important new locus whereby EPO induces substantial long-term tissue protection in the host through signaling to several critical subsets of immune cells that reside in the peripheral lymphatic system. PMID- 18382692 TI - Tumor-specific Hsp70 plasma membrane localization is enabled by the glycosphingolipid Gb3. AB - BACKGROUND: Human tumors differ from normal tissues in their capacity to present Hsp70, the major stress-inducible member of the HSP70 family, on their plasma membrane. Membrane Hsp70 has been found to serve as a prognostic indicator of overall patient survival in leukemia, lower rectal and non small cell lung carcinomas. Why tumors, but not normal cells, present Hsp70 on their cell surface and the impact of membrane Hsp70 on cancer progression remains to be elucidated. METHODOLOGY/PRINCIPAL FINDINGS: Although Hsp70 has been reported to be associated with cholesterol rich microdomains (CRMs), the partner in the plasma membrane with which Hsp70 interacts has yet to be identified. Herein, global lipid profiling demonstrates that Hsp70 membrane-positive tumors differ from their membrane-negative counterparts by containing significantly higher amounts of globotriaoslyceramide (Gb3), but not of other lipids such as lactosylceramide (LacCer), dodecasaccharideceramide (DoCer), galactosylceramide (GalCer), ceramide (Cer), or the ganglioside GM1. Apart from germinal center B cells, normal tissues are Gb3 membrane-negative. Co-localization of Hsp70 and Gb3 was selectively determined in Gb3 membrane-positive tumor cells, and these cells were also shown to bind soluble Hsp70-FITC protein from outside in a concentration-dependent manner. Given that the latter interaction can be blocked by a Gb3-specific antibody, and that the depletion of globotriaosides from tumors reduces the amount of membrane-bound Hsp70, we propose that Gb3 is a binding partner for Hsp70. The in vitro finding that Hsp70 predominantly binds to artificial liposomes containing Gb3 (PC/SM/Chol/Gb3, 17/45/33/5) confirms that Gb3 is an interaction partner for Hsp70. CONCLUSIONS/SIGNIFICANCE: These data indicate that the presence of Gb3 enables anchorage of Hsp70 in the plasma membrane of tumors and thus they might explain tumor-specific membrane localization of Hsp70. PMID- 18382693 TI - Alignment of non-covalent interactions at protein-protein interfaces. AB - BACKGROUND: The study and comparison of protein-protein interfaces is essential for the understanding of the mechanisms of interaction between proteins. While there are many methods for comparing protein structures and protein binding sites, so far no methods have been reported for comparing the geometry of non covalent interactions occurring at protein-protein interfaces. METHODOLOGY/PRINCIPAL FINDINGS: Here we present a method for aligning non covalent interactions between different protein-protein interfaces. The method aligns the vector representations of van der Waals interactions and hydrogen bonds based on their geometry. The method has been applied to a dataset which comprises a variety of protein-protein interfaces. The alignments are consistent to a large extent with the results obtained using two other complementary approaches. In addition, we apply the method to three examples of protein mimicry. The method successfully aligns respective interfaces and allows for recognizing conserved interface regions. CONCLUSIONS/SIGNIFICANCE: The Galinter method has been validated in the comparison of interfaces in which homologous subunits are involved, including cases of mimicry. The method is also applicable to comparing interfaces involving non-peptidic compounds. Galinter assists users in identifying local interface regions with similar patterns of non-covalent interactions. This is particularly relevant to the investigation of the molecular basis of interaction mimicry. PMID- 18382695 TI - A reader and author respond to "Going lean in the emergency department: a strategy for addressing emergency department overcrowding". PMID- 18382694 TI - Pharmacogenomics of interferon-beta therapy in multiple sclerosis: baseline IFN signature determines pharmacological differences between patients. AB - BACKGROUND: Multiple sclerosis (MS) is a heterogeneous disease. In order to understand the partial responsiveness to IFNbeta in Relapsing Remitting MS (RRMS) we studied the pharmacological effects of IFNbeta therapy. METHODOLOGY: Large scale gene expression profiling was performed on peripheral blood of 16 RRMS patients at baseline and one month after the start of IFNbeta therapy. Differential gene expression was analyzed by Significance Analysis of Microarrays. Subsequent expression analyses on specific genes were performed after three and six months of treatment. Peripheral blood mononuclear cells (PBMC) were isolated and stimulated in vitro with IFNbeta. Genes of interest were measured and validated by quantitative realtime PCR. An independent group of 30 RRMS patients was used for validation. PRINCIPAL FINDINGS: Pharmacogenomics revealed a marked variation in the pharmacological response to IFNbeta between patients. A total of 126 genes were upregulated in a subset of patients whereas in other patients these genes were downregulated or unchanged after one month of IFNbeta therapy. Most interestingly, we observed that the extent of the pharmacological response correlates negatively with the baseline expression of a specific set of 15 IFN response genes (R = -0.7208; p = 0.0016). The negative correlation was maintained after three (R = -0.7363; p = 0.0027) and six (R = 0.8154; p = 0.0004) months of treatment, as determined by gene expression levels of the most significant correlating gene. Similar results were obtained in an independent group of patients (n = 30; R = -0.4719; p = 0.0085). Moreover, the ex vivo results could be confirmed by in vitro stimulation of purified PBMCs at baseline with IFNbeta indicating that differential responsiveness to IFNbeta is an intrinsic feature of peripheral blood cells at baseline. CONCLUSION: These data imply that the expression levels of IFN response genes in the peripheral blood of MS patients prior to treatment could serve a role as biomarker for the differential clinical response to IFNbeta. PMID- 18382696 TI - Stroke treatment and prevention are not the same in men and women. PMID- 18382697 TI - The need for a US health television network. PMID- 18382698 TI - A 10-year-old girl with mild elevation of liver transaminases. PMID- 18382699 TI - Study of plasma adrenomedullin level in normal pregnancy and preclampsia. AB - AIM: The aim of this study was to evaluate whether maternal circulating adrenomedullin (AM) values in patients with preeclampsia are different from those in normotensive pregnant women at different gestational ages. SUBJECTS AND METHODS: In a prospective clinical study, 90 women aged 17 to 40 years old, were divided into 4 main groups: group I (45 women): Normotensive pregnant women at first trimester (15 women), second trimester (15 women), and third trimester (15 women) of pregnancies. Group II (15 women): Pregnant women with preeclampsia at 25 to 38 weeks of gestation. Group III (15 women): Normotensive healthy nonpregnant women. Group IV (15 women): Hypertensive nonpregnant women. The plasma AM concentration was measured in all women by using enzyme immunoassay kits. RESULTS: Plasma AM levels in pregnant women with normal blood pressure at different gestational ages (first, second, and third trimesters) were statistically significantly higher than those detected in nonpregnant normotensive women and significantly increased with increasing gestational age (P < .001). Moreover, there was significant positive correlation between plasma AM levels and increasing gestational age (r = 0.915, P < .001). Preeclamptic patients had the highest mean plasma AM levels compared with all other groups, which is statistically significant (P < .001) and there was a significant positive correlation between plasma AM levels and systolic blood pressure, diastolic blood pressure, severity of preeclampsia, and proteinuria in pregnant patients with preeclampsia. CONCLUSION: Maternal plasma AM concentration increases throughout pregnancy and increases as gestational age progresses. AM production starts very early in gestation, suggesting that it may have an important role in human reproduction, from implantation to delivery. Maternal plasma AM level in preeclampsia appears to be higher than that in normal pregnancy. PMID- 18382700 TI - A clinical case study: sclerosing mesenteritis presenting as chylous ascites. PMID- 18382701 TI - Part 2: A reader responds to "Getting control of health costs--for real". PMID- 18382702 TI - Spread of MRSA: past time for action. PMID- 18382703 TI - Physicians and nurses together can improve patient safety. PMID- 18382704 TI - A reader and author respond to "Reimbursement for new technologies: the GERD maze is not patient-friendly". PMID- 18382705 TI - The use of ultrasonography in the diagnosis of ectopic pregnancy: a case report and review of the literature. AB - Ectopic pregnancy is still the most common cause of maternal deaths in the first trimester. Here we report a case of a 29-year-old pregnant woman who presented with a 2-week history of lower abdominal pain and vaginal bleeding. She was initially diagnosed as having had a complete miscarriage. On the second presentation the patient was diagnosed with a tubal ectopic pregnancy. The patient was managed surgically and recovered uneventfully. The case is discussed, and a review of the literature in relation to the use of ultrasonography in the diagnosis of ectopic pregnancy is presented. PMID- 18382706 TI - Fulminant meningococcal sepsis in a woman with previously unknown hyposplenism. PMID- 18382707 TI - What is the best way to treat and rehabilitate a ruptured achilles tendon? PMID- 18382708 TI - Readers' and authors' responses to "Science and sensibility: an interview with professor Rolf M. Zinkernagel, Nobel Prize winner for medicine 1996". PMID- 18382709 TI - Navigating in the sea of healthcare. PMID- 18382711 TI - Tracheobronchial amyloidosis: utilization of radiotherapy as a treatment modality. AB - Tracheobronchial amyloidosis (TBA) is a rare disease. No general consensus exists with regard to its optimal treatment, resulting in a variety of modalities used to manage this condition. In this article, we present a case of TBA treated with external beam radiation therapy with encouraging results. A brief literature review of this rare ailment is also included. PMID- 18382710 TI - Immunohistochemical study of CD99 and EMA expression in ependymomas. AB - Tumors of the central nervous system (CNS) represent a unique, heterogeneous population of neoplasms and include both benign and malignant tumors. The present study was carried out on a total of 79 archival cases of ependymal tumors in addition to a variety of other primary CNS tumors. The study entailed the use of CD99 monoclonal antibody and epithelial membrane antigen (EMA). It was found that all 38 ependymoma cases (classic and nonclassic) showed positive membranous and intracytoplasmic CD99 immunoreactivity. Upon comparing with other CNS tumors (41 cases), it was found that CD99 could differentiate between ependymomas and nonependymal tumors, but intensity and pattern of staining were of no consequence in determining variant type or degree of histologic aggressiveness. In regard to EMA immunoreactivity, which was restricted to the ependymoma group, 2 patterns of staining could be detected--the intracytoplasmic dotlike pattern and the ringlike pattern--but some cases were completely negative. Thus, EMA was found to be of little value in the diagnosis of ependymoma and in the differentiation between different types and grades. CD99 can hence be recommended for use as a good marker for differentiation between ependymal and other CNS tumors. EMA expression and pattern of distribution, on the other hand, cannot be employed to determine the type of variant or the degree of tumor aggressiveness, and hence cannot predict the behavior of ependymal neoplasms. PMID- 18382712 TI - Fat reduction by topical waist applications may actually work. PMID- 18382713 TI - Part 3: Readers respond to "Getting control of health costs--for real": a Canadian reader and a US reader debate the validity of the Canadian Healthcare System. PMID- 18382714 TI - Clinicians must reinvent the medical record to stimulate the adoption of electronic medical records. PMID- 18382715 TI - A reader and author respond to "ePatients: engaging patients in their own care". PMID- 18382716 TI - Fibromyalgia is not a rheumatologic disease anymore. PMID- 18382717 TI - High eosinophils after HALO training: an active-duty soldier with recent travel, nonproductive cough, and eosinophilia. PMID- 18382718 TI - Personality as a prognostic factor for specialty choice: a prospective study of 4 medical school classes. AB - BACKGROUND: To augment the availability of medical care for a population that is older and more ethnically diverse, the number of US medical schools is increasing and existing medical schools are enlarging their class sizes. Predictors of specialty choice, especially primary care careers, are helpful to medical school officials and faculty involved in medical school recruitment and counseling and to students planning their career paths. OBJECTIVE: The objective was to examine the association between personality characteristics and specialty choice. METHODS: The Neuroticism-Extraversion-Openness Personality Inventory Revised (NEO PI-R) was administered to 4 Tulane University School of Medicine (New Orleans, Louisiana) classes (2003-2006). The NEO PI-R is a measure of 5 personality characteristics (neuroticism, extraversion, openness, agreeableness, and conscientiousness), with each domain having 6 underlying facets. The specialty choice of graduates was obtained from the National Residency Matching Program. RESULTS: Starting in 1999, 595 students matriculated and by June 2006, 542 (91%) had matched to residency programs in 22 specialties. There were differences among specialties for neuroticism (P = .006), openness (P < .001), and agreeableness (P = .003), but not for extraversion (P = .173) or conscientiousness (P = .103). Various pairwise differences between specialty categories were found. DISCUSSION: Eleven specialty categories were compared using the NEO PI-R. Numerous specialty variations were identified for neuroticism, openness, and agreeableness. The findings may be useful to medical school officials and faculty who recruit and counsel students and to students themselves as they reflect on their personality characteristics on their path to making career choices. PMID- 18382719 TI - Does calcium supplementation, with or without vitamin D, reduce bone loss and fractures in older adults? PMID- 18382720 TI - The top ten unfounded health scares of the year. PMID- 18382721 TI - Gerald Shulman digests his award. Interviewed by Karen Honey. PMID- 18382722 TI - Something lost--something gained: the ASCI begins its second century. AB - For the hundredth anniversary of the American Society for Clinical Investigation, we have invited various members of the community to recount here the impact that the Society, its annual meeting, and The Journal of Clinical Investigation have had on them. Their recollections provide a view of the changes that have occurred in academic medicine in general and for the physician-scientist in particular. PMID- 18382723 TI - The ASCI, the spring meetings, and growing up in academic medicine: a personal perspective. AB - For many young physician-scientists, the American Society for Clinical Investigation spring meetings are the backdrop to their initiation into academic medicine. Membership in the ASCI is a high honor and represents one's maturation and accomplishment in clinical research. The ASCI continues to provide this meeting forum for young investigators who aspire to emulate their idols and mentors just as I did in 1969 when I attended the spring meetings in Atlantic City for the first time. PMID- 18382724 TI - ASCI: reflections on the first 100 years and a proposal for the next. AB - Congratulations to the ASCI and its membership on the first 100 years. From their first organizational meeting on the Boardwalk in Atlantic City in June 1907, our predecessors recognized the need for an organization for the clinician who had a strong interest in true experimental medicine. These nine Young Turks formed the Society, had their first meeting at the Willard Hotel in Washington, DC, in 1909, and went on to become some of the leaders of American medicine in the first half of the 20th century. The establishment of the JCI in 1924 further enhanced the high standards of the ASCI and its membership. Surely thousands of us have benefited from the foundation of the ASCI and the JCI, as have, I would submit, millions of patients as well. PMID- 18382726 TI - Atlantic City memories. AB - Fifty years ago, the Atlantic City meetings, held the first week in May of every year, were attended by all the elite of American academic medicine and all who wanted to join that group. Part of the magic of those meetings was that professors and neophytes took each other seriously and talked to each other. PMID- 18382725 TI - From fatty streak to fatty liver: 33 years of joint publications in the JCI. AB - The ASCI is notable for two unique functions--the annual meeting and The Journal of Clinical Investigation. Both have inspired us over the 37 years of our collaborative adventures in research. In this retrospective, we review highlights from our 26 joint papers in the JCI, focusing on two papers that revealed the consequences of lipid accumulation with implications for atherosclerosis and steatohepatitis. PMID- 18382727 TI - The American Society for Clinical Investigation--the first 100 years. AB - The annual Atlantic City meeting of the Young Turks was an exciting event--an opportunity to hear great science, to explore career opportunities, and to meet and make friends. Though the 1950s, '60s, and '70s, the meetings remained relatively intimate, broadly covering the best in medicine and constantly growing until the participants outgrew the Atlantic City venue to eventually spawn the numerous specialty annual meetings that we have today. PMID- 18382728 TI - Homage to Atlantic City. AB - In this article, I reflect on the unique value for the societies of academic internal medicine of their annual spring meetings that were held in Atlantic City for two generations prior to 1977 and consider whether lessons remain from those past experiences. PMID- 18382729 TI - Atlantic City and the Boardwalk: 1932--1976. AB - Memories of the meetings in Atlantic City of the two major academic medical societies, the AAP and the ASCI, are enveloped by a vague and unsettling nostalgia. Dominating the scene was the Boardwalk--a site of unexpected encounters, often with long-forgotten colleagues, evoking a feeling of shared intellectual excitement and rich personal ties. PMID- 18382730 TI - The American Society for Clinical Investigation, 1952--1975: a personal perspective. AB - In this perspective, I trace my experiences with the ASCI, beginning in 1952, when as a medical student I attended my first meeting, until 1975, when I completed my term as president of the Society. I focus attention on the sociological aspects of the Atlantic City meetings and the critically important role these meetings played in the evolution of academic medicine during the third quarter of the 20th century. PMID- 18382731 TI - The old Atlantic City meetings. AB - Where once the annual meeting of the American Federation for Clinical Research, the American Society for Clinical Investigation, and the Association of American Physicians could unite the whole of clinical investigation, now stand many organizations and meetings catering to specialized fields, and the cohering effect of the Atlantic City meetings has not since been duplicated. PMID- 18382732 TI - The annual ASCI meeting: does nostalgia have a future? AB - For many academic physician-scientists, the yearly Tri-Societies meeting of the ASCI, AAP, and AFCR during the 1960s, '70s, and '80s was an annual rite of spring and the focal point of the academic year. In this brief essay, I set down some miscellaneous recollections of these meetings and some thoughts about why they were of such central importance in the careers of those of my generation. PMID- 18382733 TI - The physician-scientist: a value proposition. AB - The American Society for Clinical Investigation has supported the career development of physician-scientists for the past 100 years. As the ASCI looks to its next 100 years, it must be a leading force, not only for advancing the research of physician-scientists, but also for stimulating public advocacy for biomedical research in this country. PMID- 18382734 TI - Atlantic City is passe--I'm betting on Chicago. AB - I have news for the older generation who regale us with tales of presenting before the giants in Atlantic City. Pronouncements regarding the demise of such encounters are premature. The giants now convene in Chicago, but they are just as imposing. PMID- 18382735 TI - The birth of the American Physician Scientists Association--the next generation of Young Turks. AB - The American Society for Clinical Investigation (ASCI) was started a century ago to foster and to address the needs of the younger physician-scientists. A hundred years later, ASCI remains one of the premier organizations for physician scientists and one of most well-respected organizations in the medical community. I have had the opportunity and pleasure to interact with the ASCI not only as an organization through my tenure as president of the American Physician Scientists Association, but also with its members over the last four years. In my view, the same characteristics that permeate ASCI the organization also define ASCI the membership--mentorship, exemplary role models, advocacy, and leadership. PMID- 18382736 TI - Global health inequity: scientific challenges remain but can be solved. AB - Advances in science and technology have transformed the health of the populations of the developed world, with substantial increases in life expectancy and reductions in morbidity. These advances have not, however, touched the lives of the poorest people of the world--the billions living in developing countries. This Review Series on global health highlights the key factors contributing to inequity in health across the globe and the scientific questions that remain unanswered but are critical to creating effective and appropriate health solutions. The gaps in knowledge identified in this series point the way for scientists to contribute to a changed world. PMID- 18382737 TI - The spread, treatment, and prevention of HIV-1: evolution of a global pandemic. AB - The most up-to-date estimates demonstrate very heterogeneous spread of HIV-1, and more than 30 million people are now living with HIV-1 infection, most of them in sub-Saharan Africa. The efficiency of transmission of HIV-1 depends primarily on the concentration of the virus in the infectious host. Although treatment with antiviral agents has proven a very effective way to improve the health and survival of infected individuals, as we discuss here, the epidemic will continue to grow unless greatly improved prevention strategies can be developed and implemented. No prophylactic vaccine is on the horizon. However, several behavioral and structural strategies have made a difference--male circumcision provides substantial protection from sexually transmitted diseases, including HIV 1, and the application of antiretroviral agents for prevention holds great promise. PMID- 18382738 TI - Confronting the scientific obstacles to global control of tuberculosis. AB - Tuberculosis (TB) is a major threat to global health, recently exacerbated by the emergence of highly drug-resistant forms of the disease-causing pathogen and synergy with HIV/AIDS. In 2006, the Stop TB Partnership published "The global plan to stop TB: 2006--2015," which set out a vision of halving the prevalence of and mortality caused by the disease by 2015, followed by eliminating the disease as a public health problem by 2050. This vision depends on the development of improved diagnostics, simpler treatment, and more effective vaccination. Recently, active translational research pipelines directed toward each of these goals have been established, but improved understanding of the fundamental biology of this complex disease will prove to be the key to radical advances in TB control. PMID- 18382739 TI - Malaria: progress, perils, and prospects for eradication. AB - There are still approximately 500 million cases of malaria and 1 million deaths from malaria each year. Yet recently, malaria incidence has been dramatically reduced in some parts of Africa by increasing deployment of anti-mosquito measures and new artemisinin-containing treatments, prompting renewed calls for global eradication. However, treatment and mosquito control currently depend on too few compounds and thus are vulnerable to the emergence of compound-resistant parasites and mosquitoes. As discussed in this Review, new drugs, vaccines, and insecticides, as well as improved surveillance methods, are research priorities. Insights into parasite biology, human immunity, and vector behavior will guide efforts to translate parasite and mosquito genome sequences into novel interventions. PMID- 18382740 TI - Enteric infections, diarrhea, and their impact on function and development. AB - Enteric infections, with or without overt diarrhea, have profound effects on intestinal absorption, nutrition, and childhood development as well as on global mortality. Oral rehydration therapy has reduced the number of deaths from dehydration caused by infection with an enteric pathogen, but it has not changed the morbidity caused by such infections. This Review focuses on the interactions between enteric pathogens and human genetic determinants that alter intestinal function and inflammation and profoundly impair human health and development. We also discuss specific implications for novel approaches to interventions that are now opened by our rapidly growing molecular understanding. PMID- 18382741 TI - Pneumonia research to reduce childhood mortality in the developing world. AB - Pneumonia is an illness, usually caused by infection, in which the lungs become inflamed and congested, reducing oxygen exchange and leading to cough and breathlessness. It affects individuals of all ages but occurs most frequently in children and the elderly. Among children, pneumonia is the most common cause of death worldwide. Historically, in developed countries, deaths from pneumonia have been reduced by improvements in living conditions, air quality, and nutrition. In the developing world today, many deaths from pneumonia are also preventable by immunization or access to simple, effective treatments. However, as we highlight here, there are critical gaps in our understanding of the epidemiology, etiology, and pathophysiology of pneumonia that, if filled, could accelerate the control of pneumonia and reduce early childhood mortality. PMID- 18382742 TI - Kinetoplastids: related protozoan pathogens, different diseases. AB - Kinetoplastids are a group of flagellated protozoans that include the species Trypanosoma and Leishmania, which are human pathogens with devastating health and economic effects. The sequencing of the genomes of some of these species has highlighted their genetic relatedness and underlined differences in the diseases that they cause. As we discuss in this Review, steady progress using a combination of molecular, genetic, immunologic, and clinical approaches has substantially increased understanding of these pathogens and important aspects of the diseases that they cause. Consequently, the paths for developing additional measures to control these "neglected diseases" are becoming increasingly clear, and we believe that the opportunities for developing the drugs, diagnostics, vaccines, and other tools necessary to expand the armamentarium to combat these diseases have never been better. PMID- 18382744 TI - New challenges in studying nutrition-disease interactions in the developing world. AB - Latest estimates indicate that nutritional deficiencies account for 3 million child deaths each year in less-developed countries. Targeted nutritional interventions could therefore save millions of lives. However, such interventions require careful optimization to maximize benefit and avoid harm. Progress toward designing effective life-saving interventions is currently hampered by some serious gaps in our understanding of nutrient metabolism in humans. In this Personal Perspective, we highlight some of these gaps and make some proposals as to how improved research methods and technologies can be brought to bear on the problems of undernourished children in the developing world. PMID- 18382748 TI - Initial experience in the treatment of thoracic aortic aneurysmal disease with a thoracic aortic endograft at Baylor University Medical Center. AB - A retrospective review of 27 patients who underwent endovascular repair of thoracic aneurysms and of other thoracic aortic pathology with the thoracic aortic endograft (Gore Medical, Flagstaff, AZ) from June 2005 to July 2007 was performed. The mean follow-up period was 13.5 months (range, 2-25 months). Indications for thoracic endografting included descending thoracic aneurysms (n = 18), thoracoabdominal aneurysms (n = 3), traumatic aortic injuries (n = 3), penetrating aortic ulcers (n = 2), and contained rupture of a type B dissection (n = 1). One patient died during the procedure, for an overall mortality rate of 3.7%. The average length of stay was 8.1 days, with an average stay in the intensive care unit of 4.2 days. If patients with traumatic aortic injuries were excluded, the average overall and intensive care unit length of stay were 5.6 and 1.8 days, respectively. There was one incident of spinal cord ischemia (3.7%). There were five type I or type III endoleaks, three of which required revision (11.1%). In conclusion, thoracic endografting is a safe and viable option for the repair of descending thoracic aneurysms and other aortic pathologies. We have found it to be less invasive, even in conjunction with preoperative debranching procedures, with a shorter recovery time, decreased perioperative morbidity and blood loss, and decreased peri-operative mortality compared with standard open repair. PMID- 18382743 TI - Helminth infections: the great neglected tropical diseases. AB - Helminths are parasitic worms. They are the most common infectious agents of humans in developing countries and produce a global burden of disease that exceeds better-known conditions, including malaria and tuberculosis. As we discuss here, new insights into fundamental helminth biology are accumulating through newly completed genome projects and the nascent application of transgenesis and RNA interference technologies. At the same time, our understanding of the dynamics of the transmission of helminths and the mechanisms of the Th2-type immune responses that are induced by infection with these parasitic worms has increased markedly. Ultimately, these advances in molecular and medical helminth biology should one day translate into a new and robust pipeline of drugs, diagnostics, and vaccines for targeting parasitic worms that infect humans. PMID- 18382749 TI - Active intravascular rewarming for hypothermia associated with traumatic injury: early experience with a new technique. AB - Hypothermia is a significant contributor to mortality in severely injured patients. Rewarming is an enormous challenge, especially in those who require operative or angiographic intervention. In this patient population, external warming methods are only capable of reducing further heat loss, whereas active rewarming adds heat to the body's core but is invasive. This article analyzes our initial experience with a minimally invasive, continuous, automated, and easily portable intravascular rewarming technique using the Alsius Corporation's CoolGard system. The records of 11 hypothermic critically injured patients presenting to our level 1 trauma center over a 6-month period were reviewed. The patients' mean age was 39 +/- 22 years, 7 (64%) were male, and 7 (64%) had blunt mechanisms of injury. The mean injury severity score was 40 +/- 16, and the mean initial systolic blood pressure was 91 +/- 60 mm Hg. The mean core temperature at the initiation of rewarming was 33.6 +/- 1.0 degrees C, and the mean rewarming rate was 1.5 +/- 1.0 degrees C/h. Six patients died (55%), two of acute exsanguination and four of unsurvivable traumatic brain injuries. One patient developed a deep vein thrombosis at the femoral catheter site and experienced a nonfatal pulmonary embolus. Our experience demonstrates that active intravascular balloon-catheter rewarming represents a practical, automated technique for the immediate and continuous treatment of hypothermia in all phases of the acute care of trauma patients. PMID- 18382750 TI - Circulating tumor cells in melanoma: a review of the literature and description of a novel technique. AB - Melanoma is a prevalent and deadly disease with limited therapeutic options. Current prognostic factors are unable to adequately guide treatment. Circulating tumor cells are a disease-specific factor that can be used as a prognostic variable to guide therapy. Most research to date has focused on identification of circulating tumor cells using various methods, including polymerase chain reaction. These techniques, however, have poor sensitivity and variable specificity and predictive significance. A recently developed technology to identify circulating tumor cells is the CellSearch system. This system uses immunomagnetic cell labeling and digital microscopy. This technology may provide an alternative method to identify circulating tumor cells in patients with advanced-stage melanoma and function as a prognostic factor. We review the literature on circulating tumor cells in melanoma and present data collected at our institution using the CellSearch system in nine patients with stage III or IV melanoma. PMID- 18382745 TI - As the world grows: contraception in the 21st century. AB - Contraceptives that are readily available and acceptable are required in many poorer countries to reduce population growth and in all countries to prevent maternal morbidity and mortality arising from unintended pregnancies. Most available methods use hormonal steroids or are variations of barrier methods. Reports from several fora over the last 12 years have emphasized the number of unwanted pregnancies and resultant abortions, which indicate an unmet need for safe, acceptable, and inexpensive contraceptive methods. This unmet need can be assuaged, in part, by development of new nonhormonal contraceptive methods. This Review addresses the contribution that the "omic" revolution can make to the identification of novel contraceptive targets, as well as the progress that has been made for different target molecules under development. PMID- 18382751 TI - Current role of medical treatment and invasive management in carotid atherosclerotic disease. AB - As a significant known risk factor for the development of ischemic stroke, carotid atherosclerosis is a potentially preventable and treatable disease process. The progression of improved diagnostic modalities, including magnetic resonance and computed tomography angiography, has provided enhanced plaque detection and characterization. The management of carotid artery stenosis has also continued to evolve from an aggressive, early surgical approach with the advent of the carotid endarterectomy to the initiation of progressive medical management options and the development of advanced percutaneous intervention. Carotid endarterectomy continues to be the clear treatment of choice in symptomatic patients with >70% carotid stenosis. However, strict risk factor modification, including improved antihypertensive therapy, lipid management, smoking cessation, and antiplatelet therapy, have led to less-compelling indications for immediate surgery in asymptomatic populations. In recent years, the evolution of improved percutaneous techniques and the development and approval of carotid stents have expanded the role of intervention. Several randomized trials have studied the efficacy of carotid artery stenting versus carotid endarterectomy in asymptomatic and symptomatic patients to help define the role of invasive therapy. The primary objective of this review is to summarize the current evidence and standards for the advanced diagnostic and management strategies used in asymptomatic and symptomatic patients with carotid artery stenosis. PMID- 18382752 TI - Diagnosis of congenital unicuspid aortic valve by 64-slice cardiac computed tomography. PMID- 18382753 TI - The ectopic adrenocorticotropic hormone syndrome in carcinoid tumors. AB - Ectopic production of adrenocorticotropic hormone by carcinoid tumors is relatively uncommon and may not be recognized by physicians. This report describes a woman who had Cushing syndrome from the ectopic secretion of adrenocorticotropic hormone by a carcinoid tumor. Her cause of death was a pneumonia that may have been secondary to her untreated hypercortisolism. There are threeinstructive elements of this case: 1) the recognition of Cushing syndrome, 2) the association of Cushing syndrome with low-grade (carcinoid tumors) as well as with high-grade (small cell carcinoma) neuroendocrine tumors, and 3) the need to treat the hypercortisolism as well as the tumor. PMID- 18382754 TI - Ten years of cardiothoracic transplantation at Baylor University Medical Center and ten years of heart transplantation at the Onassis Cardiac Surgery Center. PMID- 18382755 TI - William Mark Armstrong, MD: a conversation with the editor. PMID- 18382756 TI - William Levin Sutker, MD: a conversation with the editor. PMID- 18382757 TI - Enlarged thyroid gland with normal thyroid function tests. PMID- 18382758 TI - Asymptomatic translucent dome-shaped cystic nodule on the finger. PMID- 18382759 TI - Dyspnea in a pregnant 20-year-old woman with electrocardiographic right ventricular enlargement. PMID- 18382760 TI - New drugs approved in 2007. PMID- 18382761 TI - Facts and ideas from anywhere. PMID- 18382762 TI - A novel antiplatelet antibody therapy that induces cAMP-dependent endocytosis of the GPVI/Fc receptor gamma-chain complex. AB - Platelet adhesion to vascular subendothelium, mediated in part by interactions between collagen and glycoprotein VI (GPVI) complexed with Fc receptor gamma chain, is crucial for thrombus formation. Antiplatelet therapy benefits patients with various thrombotic and ischemic diseases, but the safety and efficacy of existing treatments are limited. Recent data suggest GPVI as a promising target for a novel antiplatelet therapy, for example, GPVI-specific Abs that deplete GPVI from the surface of platelets. Here, we characterized GPVI-specific auto-Abs (YA-Abs) from the first reported patient with ongoing platelet GPVI deficiency caused by the YA-Abs. To obtain experimentally useful human GPVI-specific mAbs with characteristics similar to YA-Abs, we generated human GPVI-specific mouse mAbs and selected 2 representative mAbs, mF1201 and mF1232, whose binding to GPVI was inhibited by YA-Abs. In vitro, mF1201, but not mF1232, induced human platelet activation and GPVI shedding, and mF1232 inhibited collagen-induced human platelet aggregation. Administration of mF1201 and mF1232 to monkeys caused GPVI immunodepletion with and without both significant thrombocytopenia and GPVI shedding, respectively. When a human/mouse chimeric form of mF1232 (cF1232) was labeled with a fluorescent endocytosis probe and administered to monkeys, fluorescence increased in circulating platelets and surface GPVI was lost. Loss of platelet surface GPVI mediated by cF1232 was successfully reproduced in vitro in the presence of a cAMP-elevating agent. Thus, we have characterized cAMP dependent endocytosis of GPVI mediated by a human GPVI-specific mAb as what we believe to be a novel antiplatelet therapy. PMID- 18382763 TI - Deubiquitinating enzyme CYLD negatively regulates RANK signaling and osteoclastogenesis in mice. AB - Osteoclastogenesis is a tightly regulated biological process, and deregulation can lead to severe bone disorders such as osteoporosis. The regulation of osteoclastic signaling is incompletely understood, but ubiquitination of TNF receptor-associated factor 6 (TRAF6) has recently been shown to be important in mediating this process. We therefore investigated the role of the recently identified deubiquitinating enzyme CYLD in osteoclastogenesis and found that mice with a genetic deficiency of CYLD had aberrant osteoclast differentiation and developed severe osteoporosis. Cultured osteoclast precursors derived from CYLD deficient mice were hyperresponsive to RANKL-induced differentiation and produced more and larger osteoclasts than did controls upon stimulation. We assessed the expression pattern of CYLD and found that it was drastically upregulated during RANKL-induced differentiation of preosteoclasts. Furthermore, CYLD negatively regulated RANK signaling by inhibiting TRAF6 ubiquitination and activation of downstream signaling events. Interestingly, we found that CYLD interacted physically with the signaling adaptor p62 and thereby was recruited to TRAF6. These findings establish CYLD as a crucial negative regulator of osteoclastogenesis and suggest its involvement in the p62/TRAF6 signaling axis. PMID- 18382764 TI - The type I IFN induction pathway constrains Th17-mediated autoimmune inflammation in mice. AB - IFN-beta, a type I IFN, is widely used for the treatment of MS. However, the mechanisms behind its therapeutic efficacy are not well understood. Using a murine model of MS, EAE, we demonstrate that the Th17-mediated development of autoimmune disease is constrained by Toll-IL-1 receptor domain-containing adaptor inducing IFN-beta-dependent (TRIF-dependent) type I IFN production and its downstream signaling pathway. Mice with defects in TRIF or type I IFN receptor (IFNAR) developed more severe EAE. Notably, these mice exhibited marked CNS inflammation, as manifested by increased IL-17 production. In addition, IFNAR dependent signaling events were essential for negatively regulating Th17 development. Finally, IFN-beta-mediated IL-27 production by innate immune cells was critical for the immunoregulatory role of IFN-beta in the CNS autoimmune disease. Together, our findings not only may provide a molecular mechanism for the clinical benefits of IFN-beta in MS but also demonstrate a regulatory role for type I IFN induction and its downstream signaling pathways in limiting Th17 development and autoimmune inflammation. PMID- 18382765 TI - An antiproliferative BMP-2/PPARgamma/apoE axis in human and murine SMCs and its role in pulmonary hypertension. AB - Loss-of-function mutations in bone morphogenetic protein receptor II (BMP-RII) are linked to pulmonary arterial hypertension (PAH); the ligand for BMP-RII, BMP 2, is a negative regulator of SMC growth. Here, we report an interplay between PPARgamma and its transcriptional target apoE downstream of BMP-2 signaling. BMP 2/BMP-RII signaling prevented PDGF-BB-induced proliferation of human and murine pulmonary artery SMCs (PASMCs) by decreasing nuclear phospho-ERK and inducing DNA binding of PPARgamma that is independent of Smad1/5/8 phosphorylation. Both BMP-2 and a PPARgamma agonist stimulated production and secretion of apoE by SMCs. Using a variety of methods, including short hairpin RNAi in human PASMCs, PAH patient-derived BMP-RII mutant PASMCs, a PPARgamma antagonist, and PASMCs isolated from PPARgamma- and apoE-deficient mice, we demonstrated that the antiproliferative effect of BMP-2 was BMP-RII, PPARgamma, and apoE dependent. Furthermore, we created mice with targeted deletion of PPARgamma in SMCs and showed that they spontaneously developed PAH, as indicated by elevated RV systolic pressure, RV hypertrophy, and increased muscularization of the distal pulmonary arteries. Thus, PPARgamma-mediated events could protect against PAH, and PPARgamma agonists may reverse PAH in patients with or without BMP-RII dysfunction. PMID- 18382767 TI - Gadd45beta promotes hepatocyte survival during liver regeneration in mice by modulating JNK signaling. AB - In the liver, the JNK cascade is induced downstream of TNF receptors (TNFRs) in response to inflammatory, microbial, and toxic challenges. Sustained activation of JNK triggers programmed cell death (PCD), and hepatocyte survival during these challenges requires induction of the NF-kappaB pathway, which antagonizes this activation by upregulating target genes. Thus, modulation of JNK activity is crucial to the liver response to TNFR-mediated challenge. The basis for this modulation, however, is unknown. Here, we investigated the role of the NF-kappaB target Gadd45b in the regulation of hepatocyte fate during liver regeneration after partial hepatectomy. We generated Gadd45b(-/-) mice and found that they exhibited decreased hepatocyte proliferation and increased PCD during liver regeneration. Notably, JNK activity was markedly increased and sustained in livers of Gadd45b(-/-) mice compared with control animals after partial hepatectomy. Furthermore, imposition of a Jnk2-null mutation, attenuating JNK activity, completely rescued the regenerative response in Gadd45b(-/-) mice. Interestingly, Gadd45beta ablation did not affect hepatotoxic JNK signaling after a TNFR-mediated immune challenge, suggesting specificity in the inducible hepatic program for JNK restraint activated during distinct TNFR-mediated challenges. These data provide a basis for JNK suppression during liver regeneration and identify Gadd45beta as a potential therapeutic target in liver diseases. PMID- 18382766 TI - Acute effects of leptin require PI3K signaling in hypothalamic proopiomelanocortin neurons in mice. AB - Normal food intake and body weight homeostasis require the direct action of leptin on hypothalamic proopiomelanocortin (POMC) neurons. It has been proposed that leptin action requires PI3K activity. We therefore assessed the contribution of PI3K signaling to leptin's effects on POMC neurons and organismal energy balance. Leptin caused a rapid depolarization of POMC neurons and an increase in action potential frequency in patch-clamp recordings of hypothalamic slices. Pharmacologic inhibition of PI3K prevented this depolarization and increased POMC firing rate, indicating a PI3K-dependent mechanism of leptin action. Mice with genetically disrupted PI3K signaling in POMC cells failed to undergo POMC depolarization or increased firing frequency in response to leptin. Insulin's ability to hyperpolarize POMC neurons was also abolished in these mice. Moreover, targeted disruption of PI3K blunted the suppression of feeding elicited by central leptin administration. Despite these differences, mice with impaired PI3K signaling in POMC neurons exhibited normal long-term body weight regulation. Collectively, these results suggest that PI3K signaling in POMC neurons is essential for leptin-induced activation and insulin-induced inhibition of POMC cells and for the acute suppression of food intake elicited by leptin, but is not a major contributor to the regulation of long-term organismal energy homeostasis. PMID- 18382770 TI - Bis-phosphorus stabilised carbene complexes. AB - The stabilisation of the carbene centre in a complex can be achieved either by the metal moiety or the carbon substituents. The balance between these two stabilising effects determines the nature of the M=C bond and therefore the reactivity of the metal complexes. Introducing two vicinal phosphorus groups as substituents of the carbene centre proved to be rewarding, since depending on the coordination number of this atom different electronic properties are observed. Indeed, a sigma(3)-P atom possesses a lone pair that can interact with a carbene centre by destabilising its vacant p(pi) orbital. On the contrary a sigma(4)-P group presents low lying sigma* orbitals which can be involved in delocalising electronic density in the carbene p(pi) orbital by negative hyperconjugation. Therefore, PCP carbene complexes can exhibit either an electrophilic or nucleophilic reactivity depending on the nature of the phosphorus group and the metal centre. Carbenes complexes of early and late transition metals, but also of lanthanides are discussed in this perspective. PMID- 18382768 TI - Endothelin receptor antagonism prevents hypoxia-induced mortality and morbidity in a mouse model of sickle-cell disease. AB - Patients with sickle-cell disease (SCD) suffer from tissue damage and life threatening complications caused by vasoocclusive crisis (VOC). Endothelin receptors (ETRs) are mediators of one of the most potent vasoconstrictor pathways in mammals, but the relationship between vasoconstriction and VOC is not well understood. We report here that pharmacological inhibition of ETRs prevented hypoxia-induced acute VOC and organ damage in a mouse model of SCD. An in vivo ultrasonographic study of renal hemodynamics showed a substantial increase in endothelin-mediated vascular resistance during hypoxia/reoxygenation-induced VOC. This increase was reversed by administration of the dual ETR antagonist (ETRA) bosentan, which had pleiotropic beneficial effects in vivo. It prevented renal and pulmonary microvascular congestion, systemic inflammation, dense rbc formation, and infiltration of activated neutrophils into tissues with subsequent nitrative stress. Bosentan also prevented death of sickle-cell mice exposed to a severe hypoxic challenge. These findings in mice suggest that ETRA could be a potential new therapy for SCD, as it may prevent acute VOC and limit organ damage in sickle-cell patients. PMID- 18382769 TI - A hypomorphic mouse model of dystrophic epidermolysis bullosa reveals mechanisms of disease and response to fibroblast therapy. AB - Dystrophic epidermolysis bullosa (DEB) is a severe skin fragility disorder associated with trauma-induced blistering, progressive soft tissue scarring, and increased risk of skin cancer. DEB is caused by mutations in type VII collagen. In this study, we describe the generation of a collagen VII hypomorphic mouse that serves as an immunocompetent animal model for DEB. These mice expressed collagen VII at about 10% of normal levels, and their phenotype closely resembled characteristics of severe human DEB, including mucocutaneous blistering, nail dystrophy, and mitten deformities of the extremities. The oral blistering experienced by these mice resulted in growth retardation, and repeated blistering led to excessive induction of tissue repair, causing TGF-beta1-mediated contractile fibrosis generated by myofibroblasts and pseudosyndactyly in the extremities. Intradermal injection of WT fibroblasts resulted in neodeposition of collagen VII and functional restoration of the dermal-epidermal junction. Treated areas were also resistant to induced frictional stress. In contrast, untreated areas of the same mouse showed dermal-epidermal separation following induced stress. These data demonstrate that fibroblast-based treatment can be used to treat DEB in a mouse model and suggest that this approach may be effective in the development of clinical therapeutic regimens for patients with DEB. PMID- 18382771 TI - Unsaturated dinickel-molybdenum clusters with N-heterocyclic carbene ligands. AB - The first examples of mixed metal trinuclear clusters carrying N-heterocyclic carbene (NHC) ligands were isolated from reactions of the complexes [Ni(NHC)ClCp] [NHC = bis-(2,6-diisopropylphenyl)- or bis-(2,4,6-trimethylphenyl)-imidazol-2 ylidene] with [Mo(CO)(3)Cp](-); the unsaturated 46-electron clusters have triangular MoNi(2) cores and the reaction pathway activates usually inert Ni-Cp and Ni-NHC bonds. PMID- 18382772 TI - Palladium complexes of new bulky fluorinated diphosphines give particularly active and regioselective catalysts for hydroxycarbonylation of styrene. AB - Some new bulky fluorinated diphosphines have been prepared and converted to palladium dichloride complexes that have been found to give increased regioselectivity and activity over other Pd-diphosphine systems when used as catalysts for hydroxycarbonylation of styrene. PMID- 18382773 TI - Selective carbon-carbon bond cleavage of 2,2'-dibromotolane via photolysis of its appropriate (diphosphine)Pt0 complex in the solid state. AB - By irradiating crystals of a (diphosphine)Pt(0) complex containing 2,2' dibromotolane as ligand with sunlight, a rare example of selective carbon-carbon bond cleavage in the solid state could be observed. PMID- 18382774 TI - Different transmetallation behaviour of [M(P4HR4)] salts toward rhodium(I) and copper(I) (M = Na, K; R = Ph, Mes; Mes = 2,4,6-Me3C6H2). AB - [K(2)(P(4)Mes(4))] (1) or [Na(2)(THF)(4)(P(4)Mes(4))] (2) (Mes = 2,4,6 Me(3)C(6)H(2)) reacts with one equivalent of HCl and subsequently with 0.5 equivalents of [{RhCl(cod)}(2)] (cod = 1,5-cyclooctadiene) to give a mixture of rhodium complexes, from which [Rh(P(4)HMes(4))(cod)] (3) and the secondary product [Rh(2)(micro-P(2)HMes(2))(mu-PHMes)(cod)(2)] (4) were isolated and characterised by X-ray diffraction studies. Alternatively, the reaction of [K(2)(P(4)Ph(4))] (5) or [Na(2)(THF)(5)(P(4)Ph(4))] (6) with one equivalent of HCl and subsequently with one equivalent of [CuCl(PCyp(3))(2)] (Cyp = cyclo C(5)H(9)) gave the complex [Cu(4)(P(4)Ph(4))(2)(PH(2)Ph)(2)(PCyp(3))(2)] (7), presumably via disproportionation of the monoanion (P(4)HPh(4))(-). PMID- 18382775 TI - On the origin of paramagnetism in planar nickel(II) complexes. AB - The general rule that in Ni(II) d(8) chemistry, tetrahedral (or nearly tetrahedral) complexes have temperature dependent magnetic moments which are usually larger than the spin-only value whilst square planar complexes are diamagnetic is broken for certain Ni[P((t)Bu)(2)(O)NR](2) complexes. These have planar coordination for various alkyl groups but have the spectral and paramagnetic properties normally associated with tetrahedral systems. In contract to previous studies, density functional calculations show the unusual adoption of a high-spin rather than low-spin arrangement in the planar systems is due to the strong pi bonding of the amide group and the preference for a planar coordination is due to greater separation between the bulky nitrogen and phosphorus substituents. PMID- 18382776 TI - Towards tuning the packing and entanglement of zigzag coordination chains by terminal ligands. AB - Solvothermal reactions of trans-stilbene-4,4'-dicarboxylic acid (H(2)STDC) and zinc(ii) acetate in the presence of systematically varied terminal ligands afforded a series of supramolecular architectures with formula [Zn(STDC)(py)(2)].py (1), [Zn(STDC)(bipy)(H(2)O)].0.5py.H(2)O (2), [Zn(STDC)(biql)] (3), [Zn(STDC)(phen)].solv (solv = DMSO, 4a; DMF, 4b), where py = pyridine, bipy = 2,2'-bipyridine, biql = 2,2'-biquinoline, phen = 1,10 phenathroline. X-Ray analyses revealed that all the compounds consist of infinite 1D zigzag polymer chains. Investigations based on intermolecular interactions illustrate that the chelate terminal ligands play a critical role in determining the packing/entangling modes of the chains and the porosity of the final three dimensional architectures. In compounds 1 and 2, the weak hydrogen bonding and/or pi-pi stacking interactions assemble the parallel chains into diamond nets with four- and two-fold interpenetration, respectively. In compound 3, the hydrogen bonding and pi-pi stacking interactions collaborate to arrange the chains in two different directions, generating a 3D supramolecular architecture with high catenation. The most interesting packing occurs in 4. Extensive pi-pi stacking interactions involving the terminal and bridging ligands arrange the chains in four different directions, and the chains are hierarchically entangled to produce an unprecedented 3D microporous framework with high stability. Based on comparative investigations, the effects of the terminal and bridging ligands on the packing of zigzag chains have been discussed. The reversible guest inclusion properties of 2 and 4 have also been demonstrated. PMID- 18382777 TI - Vanadium(V) hydrazido(2-) thiolate imine alkoxide complexes. AB - The reaction of (Me(3)Si)(2)TIP with V(NNMe(2))(OAr)(3) results in the production of V(NNMe(2))(TIP)(OAr), where TIP is 2-((2 thiolatophenylimino)methylene)phenolate. The aryloxide is readily displaced by ISiMe(3) to form an insoluble iodide complex formulated as V(NNMe(2))(TIP)(I). The iodide was used to prepare three different complexes: [V(NNMe(2))(TIP)(dmpe)]I, [V(NNMe(2))(TIP)(Bu(t)bpy)][OTf], and [V(NNMe(2))(TIP)(Bu(t)bpy)][SbF(6)]. The phosphine derivative, [V(NNMe(2))(TIP)(dmpe)]I, was characterized by X-ray diffraction and shows a quite short N-N distance of 1.293(3) A indicative of a dominant isodiazene resonance form. PMID- 18382778 TI - Syntheses and structures of two chiral zincophosphite compounds: [Zn(C8H8N2)(HPO3)] and (C6H13N2)[Zn3(C6H12N2)(HPO3)3(H2PO3)]. AB - Employing achiral organic amines (2-methylbenzimidazole and 1,4 diazabicyclo[2.2.2]octane) as the structure-directing agent and ligand, two chiral zincophosphites, [Zn(C(8)H(8)N(2))(HPO(3))] 1 and (C(6)H(13)N(2))[Zn(3)(C(6)H(12)N(2))(HPO(3))(3)(H(2)PO(3))] 2, have been hydro(solvo)-thermally synthesized, crystallizing in the chiral space group P2(1)2(1)2(1). Single-crystal structural analysis reveals that compound 1 consists of alternating ZnO(3)N and HPO(3) units exhibiting a neutral one dimensional chiral chain. Interestingly, the organic ligands interact with the inorganic chain via hydrogen-bonds in hydrogen-bonded helix fashion. Compound 2, a three-dimensional chiral open framework with 16-MR channels, is formed by the distorted {Zn(3)P(4)} secondary building unit (SBU). The organic amine molecules play dual roles as both ligands and countercations. PMID- 18382779 TI - Inorganic-organic hybrid materials with different dimensions constructed from copper-fluconazole metal-organic units and Keggin polyanion clusters. AB - Inorganic-organic hybrid materials based on Keggin polyoxometalate building blocks combined with Cu(II)/Cu(I) and flexible fluconazole ligand [1-(2,4 difluorophenyl)-1,1-bis[(1H-1,2,4-triazol-1-yl)methyl]methanol] (Hfcz) have been obtained by hydrothermal methods, namely, [Cu(II)(2)(Hfcz)(4)(SiW(12)O(40))].3H(2)O (1), [Cu(II)(4)(fcz)(4)(H(2)O)(4)(SiMo(12)O(40))].6H(2)O (2), [Cu(II)(2)(fcz)(2)][Cu(II)(4)(fcz)(4)(SiW(12)O(40))][Cu(II)(2)(fcz)(2)(H(2)O)(2)( iW(12)O(40))].6H(2)O (3), (Et(3)NH)(2)[Cu(I)(2)(Hfcz)(2)(SiW(12)O(40))].2H(2)O (4), (Et(3)NH)(2)[Cu(I)(2)(Hfcz)(2)(SiW(12)O(40))].H(2)O (5) and [Cu(I)(4)(Hfcz)(4)(SiMo(12)O(40))] (6). Their structures have been determined by single-crystal X-ray diffraction analyses, and the compounds are further characterized by elemental analyses, IR spectra and thermogravimetric (TG) analyses. In 1, Cu(II) cations are bridged by fluconazole ligands to form a 3D lvt coordination polymeric network, which is connected by (SiW(12)O(40))(4-) anions to form a complicated 3D (4,6)-connected framework with the topology of (4(2).6(4))(4(6).6(7).8(2))(2). In 2, two fcz(-) anions chelate two Cu(2+) cations to form a [Cu(fcz)](2)(2+) dimer, which is bridged by (SiW(12)O(40))(4-) polyanions to generate a 2D (4,4) grid. Compound 3 is formed by three types of co crystallizing subunits including a dimer [Cu(fcz)](2)(2+), a dumbbell molecule [Cu(4)(fcz)(4)(SiW(12)O(40))] and an infinite chain {[Cu(2)(fcz)(2)(H(2)O)(2)(SiW(12)O(40))](2-)}(infinity). In compounds 4 and 5, Hfcz ligands link Cu(+) cations to generate 1D coordination polymeric units, and (SiW(12)O(40))(4-) polyanions connect these metal-organic units to form two types of (6(3)) sheets which are topological isomerism. In compound 6, (SiMo(12)O(40))(4-) polyanions fixed in Cu(I)-Hfcz square rings are further extended into a 2D sheet via linking Cu(I) atoms of different rings. By carefully inspection of the structures of 1-6, it is believed that various transition-metal organic units and Keggin polyanions with different coordination modes are important for the formation of the different structures. In addition, electrochemical behaviors of compounds 1, 2, 5 and 6 have been investigated. PMID- 18382780 TI - Mechanistic implications of the active species involved in the oxidation of hydrocarbons by iron complexes of pyrazine-2-carboxylic acid. AB - The reactivity towards H(2)O(2) of the complexes [Fe(pca)(2)(py)(2)].py (1) and Na(2){[Fe(pca(3))](2)O}.2H(2)O.CH(3)CN (2) (where pca(-) is pyrazine-2 carboxylate) and their catalytic activity in the oxidation of hydrocarbons is reported. Addition of H(2)O(2) to 1 results in the formation of a dinuclear Fe(III)-(mu-O)-Fe(III) species characterized spectroscopically and by cyclic voltammetry. By contrast, treatment of 2 with H(2)O(2) results in the formation of mononuclear iron(II) complexes, [Fe(pca)(2)(solvent)(2)]. The experimental results indicate that the catalytic activity of the starting complexes 1 and 2 is strongly dependent on the species formed in solution. PMID- 18382781 TI - Polytypism, homochirality, interpenetration, and hydrogen-bonding in transition metal (Mn(II), Ni(II), Cu(II), Zn(II)) 5-hydroxyisophthalate coordination polymers containing 4,4'-bipyridyl. AB - We report the synthesis of five coordination polymers of divalent transition metals combined with 5-hydroxyisophthalic acid (HIP) and 4,4'-bipyridyl (bipy). Mn forms two polytypic two-dimensional coordination polymers, Mn(HIP)(bipy).3H(2)O and Mn(HIP)(bipy).(1/4)bipy.2H(2)O (I and II, with ABAB and ABC stacking sequences, respectively); Ni forms a chiral hexagonal three dimensional coordination polymer with two interpenetrating trigonal sublattices exhibiting the "dual quartz" topology, Ni(HIP)(bipy)(H(2)O) (III); Cu forms a one dimensional coordination polymer containing arrays of infinite hydrogen-bonded molecular ribbons, Cu(HIP)(2)(bipy) (IV); and Zn forms a two-dimensional coordination polymer with a stair-stepped layered structure, Zn(2)(HIP)(2)(bipy)(H(2)O)(2).H(2)O (V). The M-HIP-M connections are perpendicular to the M-bipy-M connections in all structures where they are present. PMID- 18382782 TI - Polymetallic clusters of iron(III) with derivatised salicylaldoximes. AB - The synthesis and magnetic properties of the compounds [HNEt(3)][Fe(2)(OMe)(Ph sao)(2) (Ph-saoH)(2)].5MeOH (1.5MeOH), [Fe(3)O(Et sao)(O(2)CPh)(5)(MeOH)(2)].3MeOH (2.3MeOH), [Fe(4)(Me-sao)(4)(Me-saoH)(4)] (3), [HNEt(3)](2)[Fe(6)O(2)(Me-sao)(4)(SO(4))(2)(OMe)(4)(MeOH)(2)] (4), [Fe(8)O(3)(Me sao)(3)(tea)(teaH)(3)(O(2)CMe)(3)] (5), [Fe(8)O(3)(Et sao)(3)(tea)(teaH)(3)(O(2)CMe)(3)] (6), and [Fe(8)O(3)(Ph sao)(3)(tea)(teaH)(3)(O(2)CMe)(3)] (7) are reported (Me-saoH(2) is 2' hydroxyacetophenone oxime, Et-saoH(2) is 2'-hydroxypropiophenone oxime and Ph saoH(2) is 2-hydroxybenzophenone oxime). 1-7 are the first Fe(III) compounds synthesised using the derivatised salicylaldoxime ligands, R-saoH(2). 1 is prepared by treatment of Fe(2)(SO(4))(3).6H(2)O with Ph-saoH(2) in the presence of NEt(3) in MeOH; 2 prepared by treatment of Fe(ClO(4))(2).6H(2)O with Et saoH(2) and NaO(2)CPh in the presence of NEt(4)OH in MeOH; 3 prepared by treatment of Fe(ClO(4))(2).6H(2)O with Me-saoH(2) and NaO(2)CCMe(3) in the presence of NEt(4)OH in MeOH; and 4 prepared by treatment of Fe(2)(SO(4))(3).6H(2)O with Me-saoH(2) in the presence of NEt(3) in MeOH. 4 is a rare example of a polynuclear iron complex containing a coordinated SO(4)(2-) ion. Compounds 5-7 are prepared by treatment of Fe(O(2)CMe)(2) with Me-saoH(2) (5), Et-saoH(2) (6), Ph-saoH(2) (7) in the presence of H(3)tea (triethanolamine) in MeOH, and represent the largest nuclearity Fe(III) clusters containing salicyladoxime-based ligands, joining a surprisingly small family of characterised octanuclear Fe complexes. Variable temperature magnetic susceptibilty measurements of 1, 3 and 5-7 reveal all five complexes possess S = 0 spin ground states; 2 possesses an S = 1/2 spin ground state, while 4 has an S = 4 +/- 1 spin ground state. PMID- 18382783 TI - Design, synthesis and characterization of a Pt-Gd metal-organic framework containing potentially catalytically active sites. AB - The heterobimetallic metal-organic framework {[(BPDC)PtCl(2)](3)(Gd(H(2)O)(3))(2)}.5H(2)O (BPDC = 2,2'-bipyridine-5,5' dicarboxylate) has been designed and synthesized by hydrothermal methods. The new coordination polymer contains subunits of (BPDC)PtCl(2) (1) where both N atoms of the BPDC ligand are attached to a square-planar Pt(II) center. The two remaining cis coordination sites at Pt(II) are occupied by chloride ions. The final structure (2) of the polymeric network is obtained when Gd(III) ions link together the (BPDC)PtCl(2) units, which are organized in sheets, into larger blocks. These blocks are stacked along the crystallographic [010] direction and are held together by a hydrogen bonding scheme that involves carboxylate oxygen atoms and water molecules in the coordination sphere of Gd. The coordination polymer 2 can be obtained in a single-step reaction or in a two-step synthesis where the corresponding Pt complex (1) was first synthesized followed by reacting 1 with Gd(NO(3))(3).6H(2)O. In situ high temperature powder X-ray diffraction shows that the crystalline coordination polymer transforms into an anhydrous modification at 100 degrees C. This modification is stable to 350 degrees C, at which temperature the structure starts to decompose. The coordination sphere around platinum in the polymer closely resembles organometallic Pt complexes that have been previously found to catalytically or stoichiometrically activate and functionalize hydrocarbon C-H bonds in homogeneous systems. PMID- 18382784 TI - A manganese(II) coordination polymer with mixed pyrimidine-2-carboxylate and oxalate bridges: synthesis, structure, and magnetism. AB - A new two-dimensional (2D) coordination polymer with mixed pyrimidine-2 carboxylate (pymca) and oxalate (ox) bridges, [Mn(2)(pymca)(2)(ox)(H(2)O)(2)](n) (1), was synthesized by a hydrothermal reaction involving the in situ formation of the pymca and ox ligands from 2-cyanopyrimidine (pymCN), and its structure and magnetic properties were characterized. The dianionic ox and monoanionic pymca ligands are both potentially bis(chelating) and have been used to construct anionic and cationic 2D honeycomb networks [(6,3) net], respectively. In the present compound, ox assumes the usual bis(chelating) bridging mode, but pymca serves as a tridentate bridge with one of the pyrimidyl nitrogens uncoordinated. The two different bridges collaborate to generate a highly undulated 2D layer with the (6,3) net topology, and the layers are closely packed in parallel through pi-pi interactions between pyrimidyl rings and hydrogen bonds involving coordinated water molecules, oxalate oxygens, and uncoordinated pyrimidyl nitrogens. Magnetic measurements reveal that the compound is a spin-canted antiferromagnet with weak ferromagnetic transition below T(c) = 2.4 K. PMID- 18382785 TI - A hole of human immunodeficiency virus HIV/AIDS and hunger in rural Nigerian communities. PMID- 18382786 TI - Stigma. An aspect of epilepsy not to be ignored. AB - Stigma is considered to be one of the most important factors that have a negative influence on people with epilepsy (PWE) and their families. It is a global issue commonly encountered in PWE in all cultures. Stigma may have deleterious effects on the patient's life, more than epilepsy itself. It is the duty of all health professionals to try and improve the quality of life of PWE beyond seizures control, and one of the important ways to do so is by fighting stigma. Many different health professionals including physicians, surgeons, social workers, psychologists, psychiatrists, and nurses deal with PWE, and hence stigma in PWE can be encountered and dealt with by many diverse specialties. Unfortunately, this issue is addressed primarily in specialized journals like Epilepsia, Epilepsy and Behavior, Seizure, and Social Science Medicine. Professionals interested mainly in epileptology or social sciences read such journals, and we feel that this issue should be addressed in a journal targeting readers of different specialties and interests. PMID- 18382787 TI - Dose-dependent ultrastructural changes in rat cornea after oral methylphenidate administration. AB - OBJECTIVE: To investigate dose-dependent ultrastructural changes in rat cornea after oral methylphenidate Ritalin administration. METHODS: This study was conducted in the Department of Anatomy, Gazi University Faculty of Medicine, Ankara, Turkey between March and May 2005, with a total of 27 female prepubertal Wistar albino rats, divided into 3 different dose groups 5mg/kg, 10 mg/kg, 20 mg/kg, and their control groups. They were treated orally with methylphenidate, and eye tissue was removed to process for electron microscopic studies. RESULTS: We observed that all cells, and prominently basal cells of the corneal epithelium show dose-dependent degenerative changes such as apoptotic bodies, chromatin condensation, and ondulation in their nuclei and crystolysis of the mitochondrion. In the stroma, the most evident finding was the increase of the collagen fiber. In addition to dose-dependent changes related to the apoptotic process, which is chromatin condensation in their nuclei, electron dense material accumulation, and pericellular edema in the cytoplasm were also seen. In the endothelial cell lines, disruption of the junctional complexes, vacuolization in the cell cytoplasms, and crystolysis of the mitochondrion's with rough endoplasmic reticulum cisternae activity were observed. CONCLUSION: Ritalin is inducing an evident degeneration, especially in epithelium cells with increasing doses. Ultrastructural cell organelle composition degeneration with stromal fibrosis has a negative effect on cornea dehydration. In light of these findings, we believe that the Ritalin treatment doses need to be kept to a minimum to maintain healthy cornea ultrastructure and related physiology. PMID- 18382788 TI - Study of oxidative stress in type 2 diabetic patients and its relationship with glycated hemoglobin. AB - OBJECTIVE: To evaluate the activity of antioxidant enzymes in diabetic patients and also to determine the correlation between hyperglycemia and lipid peroxidation. METHODS: Thirty patients with type 2 diabetes and 30 healthy individuals control group participated in this case-control study. The patients were referred to Sina Hospital, Hamadan, Iran from April to June 2006. Glycated hemoglobin HbA1c was measured as a marker of hyperglycemia using the chromatography method Biosystem and malondialdehyde MDA was determined using the colorimetric method. Glutathione peroxidase GPx and superoxide dismutase SOD activity were assessed using the UV-Vis spectrophotometric technique Randox kit. RESULTS: The mean of HbA1c was higher in diabetic patients compared to the healthy group, and the difference was statistically significant p<0.001. Serum MDA in diabetics was higher compared to those of healthy subjects p<0.001. There were significant differences in activities of SOD and GPx between the 2 studied groups indicating lower activity in diabetic patients p<0.001. There was a significant relationship between MDA and HbA1c in diabetic and healthy subjects. CONCLUSION: The data showed an increase in lipid peroxidation and oxidative stress in diabetes and also indicated a positive correlation between the degree of hyperglycemia and oxidative stress. Evaluation of oxidative status and choosing the appropriate treatment may help to support antioxidant defense in these patients. PMID- 18382789 TI - Gene expression profiling in women with breast cancer in a Saudi population. AB - OBJECTIVE: To generate consensus gene expression profiles of invasive breast tumors from a small cohort of Saudi females, and to explore the possibility that they may be broadly conserved between Caucasian and Middle Eastern populations. METHODS: This study was performed at King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia, from January 2005 to January 2007. Gene expression profiles were generated from 38 invasive breast tumors, and 8 tumor adjacent tissues TATs using BD Atlas cDNA expression arrays containing 1176 genes. Results were confirmed by reverse transcriptase polymerase chain reaction, and analyzed by 2-dimensional unsupervised hierarchical clustering. RESULTS: The analysis identified 48 differentially expressed genes in tumors from which 25 are already reported by various western studies. Forty-three of these genes were also differentially expressed in TATs. The same data set has been able to distinguish between tumors and the TATs, interestingly by using only 4 of the differentially expressed genes. Moreover, we were able to group the patients according to prognosis to an extent by hierarchical clustering. CONCLUSION: Our results indicate that expression profiles between Saudi females with breast cancer and the Caucasian population are conserved to some extent, and can be used to classify patients according to prognostic groups. We also suggest 3 differentially-expressed genes IGHG3, CDK6, and RPS9 in tumors may have a novel role in breast cancer. In addition, the role of TATs is much more essential in breast cancer, and needs to be explored thoroughly. PMID- 18382790 TI - Campylobacter jejuni induces diverse kinetics and profiles of cytokine genes in INT-407 cells. AB - OBJECTIVE: To examine the kinetic ability of embryonic human epithelial INT-407 cells to express messenger ribonucleic acid mRNA for various cytokines and chemokines in response to Campylobacter jejuni C. jejuni stimulation. METHODS: In an experimental single-blind study, cultured embryonic human epithelial INT-407 cells were treated with different concentrations of viable C. jejuni, its sonicated, and filtered supernatant. A modified non-radioactive in situ hybridization using probe cocktails was used to measure mRNA levels for the pro inflammatory cytokines interleukin IL-1beta, IL-6, interferon-gamma IFN-gamma, tumour necrosis factor TNF-alpha, transforming growth factor TGF-beta1, and IL-8, and the anti-inflammatory cytokines, IL-4 and IL-10. The study was carried out from September 2005 to March 2007 at the Department of Microbiology, Immunology, and Infectious Diseases, College of Medicine, Arabian Gulf University, Bahrain. RESULTS: Viable C. jejuni, sonicated bacteria and filtered supernatant induced high mRNA expression for the pro-inflammatory cytokines IL-1beta, IL-6, IFN gamma, TNF-alpha, TGF-beta1, and IL-8, which peaked at the 12 hours post stimulation. Anti-inflammatory cytokines IL-4 and IL-10 mRNA expression were induced maximally at 3 hours post stimulation mainly by sonicated bacteria and filtrated supernatant, however, not with living bacteria. Untreated embryonic human epithelial INT-407 cells expressed low amount of mRNA for the various cytokines and chemokines at all time points. For each cytokine, 4 samples were used per time hour. CONCLUSION: This study demonstrated that embryonic human epithelial INT-407 cells in response to viable C. jejuni or its cytotoxins can alter cytokine and chemokine mRNA expression patterns and kinetics suggesting a potential role for theses mediators in the immunopathogenesis of the infection caused by this pathogen, which might be relevant for future immunotherapeutic interventions during severe bacterial infections. PMID- 18382791 TI - Usefulness of procalcitonin and some inflammatory parameters in septic patients. AB - OBJECTIVE: To evaluate the efficacy of procalcitonin PCT to identify critically ill patients with sepsis in comparison with leukocyte count, body temperature, C reactive protein CRP, erythrocyte sedimentation rate ESR, and interleukin-6 IL-6. METHODS: We performed our prospective observational study in 75 patients admitted with acute systemic inflammatory response and suspected infection. The final diagnosis was systemic inflammatory response syndrome SIRS in 38 patients, sepsis in 22, severe sepsis in 10, and suspected viral sepsis in 5. Blood samples were taken on the first day of hospitalization in Al Mwasaa Hospital, Damascus, Syrian Arab Republic, from July 2006 to January 2007. We estimated the relevance of the different parameters by using the t-test, Pearson's correlation coefficient, and area under the receiver operating characteristic curves. RESULTS: Mean PCT concentrations on admission were 0.37 ng/ml for SIRS n=38, 3.31 ng/ml for sepsis n=22, 40.2 ng/ml for severe sepsis n=10, and significant differences existed in plasma PCT levels among the 3 groups. The PCT was the only distinguisher between sepsis and non-infectious SIRS, whereas it exhibited the best discriminative power between sepsis and severe sepsis with an area under the curve AUC of 0.966 followed by IL-6 with an AUC of 0.836. The PCT also do not correlate with any of the studied parameters within the SIRS group and the sepsis group. CONCLUSION: Assessing PCT levels is a more reliable way to indicate sepsis in newly admitted patients with systemic inflammations compared with conventional inflammatory parameters and IL-6. PMID- 18382792 TI - Investigation of cryptosporidiosis by enzyme-linked immunosorbent assay and microscopy in children with diarrhea. AB - OBJECTIVE: To investigate the frequency of cryptosporidiosis by enzyme-linked immunosorbent assay ELISA and microscopy and its relationship with diarrhea. METHODS: The study was prospectively performed in the Parasitology Laboratory and Pediatric Outpatient Clinic of the Research Hospital, Yuzuncu Yil University, Van, Turkey between 2004 and 2006. Stool samples were obtained from a total of 2000 children with diarrhea, 870 females, and 1130 males aging 0-15 years as study group, and 100 children of the same age were randomly selected as a control group. The flotation method was firstly carried out for all stool samples in saturated zinc sulfate solution, then staining process by modified acid-fast staining. All samples were also tested for Cryptosporidium parvum antigen by ELISA. Native-Lugol and trichrome staining were used to identify other intestinal parasites. RESULTS: The antigen was determined in 97 4.9% of 2000 children by ELISA, however, the oocysts were only seen in 39 children 1.95% by microscopy. Cryptosporidium spp. were not detected in the control group either by ELISA or by microscopy. We found a significant p<0.001 relationship between diarrhea and cryptosporidiosis. Other intestinal parasites were detected in 713 35.7% of 2000 diarrheic children. CONCLUSION: Cryptosporidium spp. antigen searching by ELISA in stool samples should be included for diagnosis of the disease in all hospitals. PMID- 18382793 TI - Comparison of 4 laboratory methods for detection of Helicobacter pylori. AB - OBJECTIVE: To compare the usefulness of 4 commonly used tests in the diagnosis of Helicobacter pylori H. pylori infection in Saudi patients with dyspepsia. METHODS: Patients presenting with dyspepsia at the gastroenterology clinic of Aseer Central Hospital, Abha, Kingdom of Saudi Arabia between October 2005 to May 2006, who consented to participate in the study were enrolled. Patients who received anti-Helicobacter treatment or proton pump inhibitors within 30 days, or in whom endoscopy revealed cancer or gastro-esophageal reflux, were excluded from the study. Stool sample for H. pylori stool antigen HpSA were tested using the enzyme immunoassay technique 2-7 days before endoscopy. At endoscopy, gastric antrum mucosal biopsies were taken for campylobacter-like organism CLO test, histology and culture. RESULTS: There were 72 males and 43 females, age range from 18-75 years, mean age 40.09+/-15.68. Seventy-nine patients out of the 115 68.7% samples were positive for H. pylori, by culture. Culture and histology agreed in 112 cases 97.4% and disagreed in 3 cases 2.6%. The sensitivities and specificities % of histology were 97.5 and 97.2, of HpSAg were 91.9 and 98.6, and of CLO were 79.7 and 97.2 tests against culture. CONCLUSION: Culture, histology, and H. pylori stool antigen tests all have comparable results, and there is no need to use all 3 at the same time, for the diagnosis of H. pylori infection. The CLO test is less sensitive, and of low negative predictive value. PMID- 18382794 TI - Cadaveric liver transplant from older donors. AB - OBJECTIVE: To examine the effects of cadaveric donor age on outcomes following orthotopic liver transplantation OLT. METHODS: Data were collected on all patients who underwent OLT between January 1997 and December 2004 at the Royal Prince Alfred Hospital, Sydney, New South Wales, Australia. During this period, 313 OLTs were performed: 51 patients 16% received older donor livers OD; 60 or more years old, and 262 84% received younger donor livers YD; less than 60 years old. RESULTS: In the study group 313 patients, we found significantly more recipients of OD liver with blood group O: 51% versus 33% p=0.025 and with fulminant hepatic failure: 9.8% versus 5% p=0.018 compared to YD recipients. No difference between OD and YD liver recipients was found in initial poor graft function: 16/51 31% versus 74/262 28%, primary non-functioning: 6.5% versus 6.5%, the overall graft loss: 15/51 29% versus 62/262 24%, post-revascularization liver biopsy steatosis: 14/40 35% versus 82/232 36% or hepatic artery thrombosis: 1/51 2% versus 8/262 3%. There was no difference in graft actuarial survival between OD and YD recipients at 1, 3, and 5 years, 82% versus 87%, 75% versus 81%, and 75% versus 77% p=0.27 log rank or patient actuarial survival, 86% versus 89%, 79% versus 83%, and 79% versus 80% p=0.336 log rank. CONCLUSION: Orthotopic liver transplantation can be achieved with acceptable outcomes using selected livers from older deceased donors. PMID- 18382795 TI - Effects of octreotide on healing of mechanical ileus in rats. AB - OBJECTIVE: To investigate the effects of octreotide on mechanical ileus without surgical intervention. METHODS: This study was organized to investigate the effects of octreotide Sandostatine one microgram/ml- Sandoz, a long acting analogue of somatostatin, in a mechanical ileus model in Sprague-Dowley rats. The study was performed at Ankara Oncology Hospital, Ankara, Turkey in April 2005. The animals were randomly separated into 3 groups. 1 control group n:10 2 ileus group n:12, and 3 ileus + octreotide group n:13. Intraluminal liquid volume, as well as concentrations of sodium, chlorine, calcium, and phosphorous were measured in systemic circulation, proximal, and middle bowel segments. We also evaluated the leukocyte count, and hematocrit levels in all animals. RESULTS: There was no difference in leukocyte count and hematocrit levels. The effects of octreotide in decreasing intestinal liquid volume, and electrolytes were significant. However, these effects were not beneficial on the mortality rates, and intestinal edema (p>0.05). CONCLUSION: We believe additional animal studies, and subsequently controlled clinical trials are necessary before using octreotide as a clinical adjunct in the treatment of small bowel obstruction. PMID- 18382796 TI - Remifentanil versus fentanyl in combination with midazolam for retrobulbar block in cataract surgery. AB - OBJECTIVE: To compare the effects of fentanyl or remifentanil in combination with midazolam on hemodynamic parameters, pain, and satisfaction profile in cataract surgery. METHODS: This randomized, double blind, prospective study was conducted between 10 and 20th July 2005 at Kudret Eye Hospital, Ankara, Turkey. Patients scheduled for cataract surgery by the phacoemulsification technique were randomly enrolled to receive sedation with midazolam 1 mg intravenous iv either with fentanyl 25 microgram group 1, n=54 or remifentanil 0.3 microgram/kg group 2, n= 46. Heart rate, systolic and diastolic arterial pressure values were recorded as baseline, after retrobulbar injection, and during the operation. We evaluated recall of retrobulbar block, pain during injection and operation, satisfaction of patient and surgeon, and the adverse effects. RESULTS: There were statistically significant alterations in systolic and diastolic arterial pressure measurements within and between groups, whereas all kept in the clinically normal range. Twenty-four percent of patients in group 1 and 15.2% in group 2 did not even remember the retrobulbar injection. The pain scores during retrobulbar injection and operation were similar in both groups. Also, satisfaction of patients and surgeon was high and comparable between groups. CONCLUSION: Remifentanil and fentanyl are both efficient and comparable opioid adjuncts to midazolam providing low injection pain and high satisfaction level with hemodynamic stability in cataract surgery under retrobulbar injection. PMID- 18382797 TI - Henoch-Schonlein purpura in children. Influence of age on the incidence of nephritis and arthritis. AB - OBJECTIVE: To assess the epidemiological, clinical, and laboratory findings in children affected with Henoch-Schonlein purpura HSP, and to analyze the influence of age on the incidence of nephritis and arthritis. METHODS: This is a retrospective study of the case records of all patients admitted to Prince Rahmah Hospital, Irbid, Jordan with HSP from September 1997 to December 2006, and to King Abdullah University Hospital, Irbid, Jordan from August 2005 to December 2006. RESULTS: The clinical manifestations included purpura in 100% of the 68 patients, arthritis in 75%, and nephritis in 29%. The incidence of nephritis varied according to the age of patients. It occurred in 19% of children less than 5 years of age, and in 67% of children 10 years of age and above. Henoch Schonlein purpura nephritis was mild in all age groups. The incidence of arthritis also varied according to the age of patients. Arthritis occurred in 56% of children less than 5 years of age, and in 89% of children 10 years of age and above. CONCLUSION: The incidence of nephritis and arthritis in children affected with HSP is age related. Older children have a higher incidence of nephritis and arthritis, while younger children have a lower incidence of nephritis and arthritis. Renal involvement was mild during the acute phase of the disease in all age groups. PMID- 18382798 TI - Painkilling effect of ozone-oxygen injection on spine and joint osteoarthritis. AB - OBJECTIVE: To analyze the painkilling effect of ozone-oxygen injection on joint and spine osteoarthritis. METHODS: This prospective study was completed at the Ozone Clinic, Rashid Hospital, Dubai, United Arab Emirates on 220 mainly local patients 122 women, mean age 47.05 years; 98 men, mean age 52.8 years with radiographic documented spine or extremities osteoarthritis. The patients were treated over 3 years September 2002 to August 2005 by ozone-oxygen injection twice a week for at least 12 sessions. Using the 6 faces pain scale; the patients' pain was recorded at the beginning and at the 4th, 8th, and 12th sessions. They were followed for a mean of 8.48 months and their pain scale was recorded at that time too. RESULTS: Comparison of the patients' 1st day pains with their 4th, 8th, and 12th sessions' pains showed a significant decrease 1st day to 4th session p=0.005, 1st day to 8th week p=0.005, 1st day to 12th session p=0.0043. Comparison of the 1st day pain with the final follow-up pain, which was around 10 months from the first treatment, showed a meaningful decrease of pain p=0.0048. CONCLUSION: This study validates the painkilling effect of ozone-oxygen injection on osteoarthritis of the joints and spine. Its long term effect on pain advocates the likelihood of some histological changes as mechanism of its action. PMID- 18382800 TI - Intraoperative assessment of the node-negative neck with frozen section biopsy. AB - OBJECTIVE: To determine the accuracy of intraoperative assessment of the node negative N0 neck with frozen-section biopsy. METHODS: In our study, 60 patients 9 women and 51 men who underwent 88 elective neck dissections with head and neck squamous cell carcinoma, in the Department of Otorhinolaryngology Medical Faculty, Cukurova University, Adana, Turkey, from January 2003 to December 2006, were reviewed retrospectively. RESULTS: Of the 88 clinically preoperatively staged N0 necks, 22 necks 25% were shown subsequently to have metastatic disease after final pathological assessment. In 16 out of 22 cases, frozen-section biopsy revealed metastatic disease, and surgery was changed to a modified comprehensive neck dissection. In another 6 cases, the sampled lymph nodes were considered negative at frozen-section analysis, and metastatic disease was found in the final histopathologic reports. There was no false-positive result. The specificity and positive predictive values were both 100%. The sensitivity of intraoperative lymph node assessment was 73%, and the negative predictive value was 91%. CONCLUSION: Intraoperative assessment of the N0 neck with frozen-section biopsy seems to improve the accuracy of staging and may alter the intraoperative treatment strategy. PMID- 18382799 TI - Apoptosis is the most efficient death-pathway in tumor cells after topoisomerase II inhibition. AB - OBJECTIVE: To compare the efficiency of apoptosis and other modes of cell death in killing tumor cells after the induction of DNA damage by topoisomerase inhibitors like etoposide. METHODS: This study was carried out in the Tumor Biology Department, National Cancer Institute, Cairo University, Cairo, Egypt, from September 2005 to August 2007. The breast cancer MCF7, the cervix carcinoma, human cervical adenocarcinoma Hela, and the brain tumor U251 cell lines were exposed to etoposide. Apoptosis was detected using the flow cytometry and the DNA ladder formation methods. Cell viability was determined by a colorimetric assay, and the residual DNA double-strand breaks dsb were measured by gel electrophoresis. RESULTS: The Hela cells were the most, the MCF7's were moderately, whereas the U251's were the least sensitive to etoposide. Apoptosis was detected only in Hela cells whereas the other 2 cell lines showed a very low level of apoptosis only 3% increase above the control cells. At equitoxic drug concentrations namely IC50, the Hela cells showed the lowest amount of non repaired DNA dsb, and the MCF7's showed the highest amount, whereas the U251 cells showed a moderate amount. CONCLUSION: These results indicate that although other modes of cell death exist, apoptosis is the most efficient and requires lower drug concentrations and fewer numbers of non-repaired dsb to give the same killing effect. Clinically, this means that tumors that can execute apoptosis may require lower doses of topoisomerase inhibitors than those that lost the ability to exercise apoptosis. PMID- 18382801 TI - The sympathetic skin response in diabetic neuropathy and its relationship to autonomic symptoms. AB - OBJECTIVE: To examine the utility of the sympathetic skin response SSR as a measure of impaired autonomic function among diabetic patients in Saudi Arabia. METHODS: In this case-control study, baseline SSR was obtained from 18 healthy subjects, followed by nerve conduction studies, and SSR testing on a consecutive cohort of 50 diabetic patients with peripheral neuropathy. The SSR in diabetic patients was compared between those with autonomic neuropathy and those without autonomic neuropathy. This study was conducted at the King Khaled University Hospital, Riyadh, Saudi Arabia, from June 2006 to June 2007. RESULTS: The SSR was present in all healthy subjects, and in 32 diabetic patients. Among 16 patients with autonomic neuropathy, the SSR was absent in 14 and present in 2, while 4 of 34 patients lacking evidence of autonomic neuropathy had absent SSR. Using Fisher's exact test, we found a strong association between absent SSR and autonomic neuropathy p<0.001, however, not with age or duration of diabetes mellitus. As a diagnostic test of autonomic neuropathy, the SSR had a sensitivity of 87.5%, a specificity of 88.2%, a positive predictive value of 77.8%, and a negative predictive value of 93.7%. CONCLUSION: Absence of the SSR is a reliable indicator of autonomic neuropathy among patients with diabetes mellitus in Saudi Arabia. PMID- 18382802 TI - Protective role of intracoronary shunt in off-pump coronary bypass operations. AB - OBJECTIVE: To investigate if there are any advantages in using intracoronary shunts compared to shuntless operations, in the context of whether it has a protective role for the myocardia. METHODS: This prospective study, included 100 patients who underwent off-pump coronary bypass surgery at 2 different cardiovascular surgery departments, namely, the Social Security Ankara Ihtisas Hospital, and Hacettepe University Hospital, Turkey, between September 2002 and July 2006. Patients were divided into 2 groups. In group 1 n=50 off-pump coronary bypass operations were performed with intracoronary shunts. In group 2 n=50 shunts were not used during off-pump. Serum creatine kinase, myoglobin, and troponin were studied. RESULTS: There were significant increases in serum creatine kinase levels in group 2 at postoperative 6th, 12th, and 24th hours. In group 2, the increase of myoglobin was statistically significant at only the postoperative 24th hour. Troponin levels were significantly higher in group 2 at postoperative 6th, 12th, and 24th hours. CONCLUSION: There are some questions regarding myocardial protection while maintaining a bloodless secure surgical field in off-pump coronary surgery. However, use of intracoronary shunts provides distal coronary flow, and reduces the risk of myocardial ischemia, while maintaining a comfortable blood free anastomosis area. PMID- 18382803 TI - An experience with a rare diagnosis of isolated tuberculosis of sternum at JNMC Hospital, Aligarh, India. AB - OBJECTIVE: To diagnose isolated tuberculosis of the sternum in patients presenting with signs and symptoms suggestive of tuberculosis. METHODS: A prospective study conducted at Jawaharlal Nehru Medical College JNMC Hospital, Aligarh, India from July 2000 to July 2006 of 2512 patients presenting with a suspected case of tuberculosis. Cases were confirmed using different investigations along with treatment of the patients with antitubercular treatment ATT, with further follow up for next 2 years. RESULTS: Nineteen patients were suffering from isolated tuberculosis of sternum. The main presenting feature was retrosternal discomfort 100% while 14 73.6% patients presented with a cold abscess of sternum. All the patients were treated successfully with 12 months of ATT and followed up for 2 years for any relapse. CONCLUSION: Although tuberculosis of sternum is rare, it should be considered as an important differential diagnosis in patients presenting with chronic lesions of the sternum, particularly in endemic/developing countries. Twelve months of treatment with ATT is optimal for achieving cure without any risk of relapse. PMID- 18382804 TI - Circulatory neutrophil chemokines in statin-treated diabetic patients. AB - OBJECTIVE: To assess the effect of statins on the circulatory levels of neutrophil chemokines, namely, granulocyte chemotactic protein-2 GCP-2, growth regulated oncogene-alpha GRO-alpha and epithelial-cell-derived neutrophil activating peptide-78 ENA-78 in patients with diabetes. METHODS: We studied 91 diabetic patients 46 were statin-treated and 45 were not and 28 healthy subjects. We measured the levels of GCP-2, GRO-alpha, and ENA-78 in the serum for the 3 groups using an enzyme linked immunosorbent assay. This cross-sectional study was conducted at King Khalid University Hospital KKUH, Riyadh, Kingdom of Saudi Arabia, from January 2006 to July 2007. RESULTS: Circulating levels of GCP-2, ENA 78, and not GRO-alpha, were significantly higher in diabetic patients as compared to healthy subjects p<0.05. Statins dropped the levels of both GCP-2 and GRO-a. The ENA-78 levels were not affected by statin therapy. There was no correlation between the levels of these chemokines with the body mass index and glycemia in the population studied. CONCLUSION: Diabetes is associated with an elevation of GCP-2 and ENA-78, and not GRO-alpha. Statins have a significant role in reducing the level of GCP-2. PMID- 18382805 TI - Bacille Calmette-Guerin immunotherapy of viral warts. AB - OBJECTIVE: To assess the effectiveness of the Bacille Calmette-Guerin BCG vaccine in the treatment of viral warts. METHODS: A single blind placebo controlled study conducted at the Department of Dermatology, Baghdad Teaching Hospital, Baghdad, Iraq from March 2005 to June 2006. Two hundred patients with viral warts were enrolled in this study, and were fully assessed before therapy. The patients were divided into 2 equal matched groups. Group 1 was designed as the treatment group and received BCG vaccine in 1-3 doses with a one-month interval, while group 2 was injected with distilled water. The patients were evaluated every 2 weeks for evidence of regression of lesions. The follow up period lasted for 3 months after the last dose. RESULTS: A total of 154 patients completed the study: 81 patients in group I and 73 patients in group II. Thirty 39.7% patients out of 81 patients in group I showed complete recovery. These results were significantly high p<0.05 when compared with total response, 10 13.7% out of 73 patients in group II. No recurrence has been reported during the follow up period. When cases that failed to respond to placebo were treated with BCG in a similar manner to group I, there was total response in 15 42.7% out of 44 patients. No side effects were observed. CONCLUSION: The BCG vaccine was an effective and safe modality of treatment of viral warts. PMID- 18382807 TI - Congenital plexiform schwannoma of the clitoris. AB - Schwannomas are slow-growing nerve sheet neoplasms which are rarely found in the female genital system. In this article, we present a patient with Antony A congenital plexiform schwannoma of the clitoris. A 6-year-old girl was brought to our hospital with the history of a firm non-cystic clitoral mass from birth, which had been growing more rapidly during the previous year. The patient was scheduled for surgery. Histological studies revealed plexiform schwannoma of the clitoris post-operatively. Therefore, schwannoma should be considered in the differential diagnosis of clitoral masses. PMID- 18382806 TI - Detection of IgG anti-beta2 glycoprotein-I antibodies in Saudi patients with systemic lupus erythematosis. AB - OBJECTIVE: To develop an assay for the measurement of this anti-human beta2 glycoprotein I (abeta2-GPI) METHODS: This study was conducted from September 2004 to December 2006. The patients attending the Rheumatology Clinic were chosen from several centers in the Eastern region of Saudi Arabia because they had complications. An enzyme-linked immunosorbent ELISA assay was optimized and developed to measure IgG abeta2-GPI antibody levels in humans. Fifty normal blood donors and 50 systemic lupus erythematosis SLE patients were selected for this experiment. RESULTS: Raised IgG abeta2-GPI antibody levels were found in 80% of SLE patients. Interestingly, raised IgG a-beta2-GPI antibody levels were associated with the presence of venous thrombosis and thrombocytopenia. CONCLUSION: The real value of IgG abeta2-GPI as a predictor for the future clinical complications needs to be confirmed in prospective controlled studies investigating clinical complications in relationship to IgG abeta2-GPI and to other risk factors for thrombosis. PMID- 18382808 TI - Unusual case of left iliac vein compression secondary to May-Thurner syndrome and crossed fused renal ectopia. AB - External compression of the left iliac vein against the fifth lumbar vertebra by the right iliac artery May and Thurner syndrome is a well-known anatomic variant. We identified a rare case of May-Thurner syndrome associated with crossed fused renal ectopia on the left side. The patient presented with complete thrombosis of the left common iliac vein down to the popliteal vein. He was treated with catheter directed thrombolysis followed by anticoagulant therapy. PMID- 18382809 TI - Primary omental pregnancy. AB - Omental pregnancy is a rare form of ectopic pregnancy, and can be seen primarily or secondary to a tubal pregnancy. A 25-year-old woman presented with abdominal distention with pain and anemia without vaginal bleeding. After a provisional diagnosis of ruptured ectopic pregnancy, laparotomy was performed. On surgical exploration, the bilateral tubes and ovaries were intact, however, an omental pregnancy was detected as the cause of hemoperitoneum. Partial omentectomy was performed. Although most cases are secondary, presented here is an additional case of primary omental pregnancy at 12 weeks according to Studdiford's criteria. Histological evidence of neovascularization into the supporting tissue confirmed our diagnosis. A primary omental pregnancy should always be considered as a possible explanation for severe hemoperitoneum in ectopic pregnancies presenting with acute abdomen, and with intact adnexes on surgical exploration. PMID- 18382810 TI - Accent method of voice therapy for treatment of severe muscle tension dysphonia. AB - Nonorganic dysphonia may present a challenging diagnosis, and management. Here, we present a severe form of nonorganic dysphonia, which we termed as arytenoidal dysphonia. It was a severe form of muscle tension dysphonia, which was described earlier in literature although with different nomenclature. The outcome of the accent method of voice therapy was also presented. We concluded that accent method of voice therapy is proven to be an effective treatment modality of arytenoidal dystonia. PMID- 18382811 TI - Intestinal perforation by multiple ectopic pancreatic tissues in a neonate with multiple congenital anomalies. AB - Ectopic pancreatic tissues of the gut are usually found incidentally during laparotomy or are reported in the autopsy findings. Rarely these ectopic pancreatic tissues may cause symptoms such as hemorrhage, pancreatitis, intussusception or perforation. We present a case report of the presence of multiple ectopic pancreatic tissues in the gut causing hemorrhage and perforation in a preterm, extremely low birth weight neonate with multiple congenital anomalies. PMID- 18382812 TI - Large solitary fibrous tumor arising from the omentum. PMID- 18382813 TI - Sensitivity of in-house polymerase chain reaction for detecting hepatitis B-DNA in HbsAg positive sera. PMID- 18382814 TI - Microbial yield in febrile sickle cell disease patients with acute painful episode from a University Hospital in the Sultanate of Oman. PMID- 18382815 TI - Emergence of multi and pan-drug resistance Acinetobacter baumannii carrying blaOXA-type -carbapenemase genes among burn patients in Tehran, Iran. PMID- 18382816 TI - Dietary intervention versus metformin to improve the reproductive outcome in women with polycystic ovarian syndrome. PMID- 18382817 TI - Evaluation of the effect of Islamic fasting on lung volumes and capacities in healthy persons. PMID- 18382818 TI - Chronic fatigue syndrome: an approach combining self-management with graded exercise to avoid exacerbations. AB - Controversy regarding the aetiology and treatment of patients with chronic fatigue syndrome continues among the medical professions. The Cochrane Collaboration advises practitioners to implement graded exercise therapy for patients with chronic fatigue syndrome using cognitive behavioural principles. Conversely, there is evidence that exercise can exacerbate symptoms in chronic fatigue syndrome, if too-vigorous exercise/activity promotes immune dysfunction, which in turn increases symptoms. When designing and implementing an exercise programme for chronic fatigue syndrome it is important to be aware of both of these seemingly opposing viewpoints in order to deliver a programme with no detrimental effects on the pathophysiology of the condition. Using evidence from both the biological and clinical sciences, this paper explains that graded exercise therapy for people with chronic fatigue syndrome can be undertaken safely with no detrimental effects on the immune system. Exercise programmes should be designed to cater for individual physical capabilities and should take into account the fluctuating nature of symptoms. In line with cognitive behaviourally and graded exercise-based strategies, self-management for people with chronic fatigue syndrome involves encouraging patients to pace their activities and respect their physical and mental limitations, with the ultimate aim of improving their everyday functioning. PMID- 18382819 TI - Eight months of physical training in warm water improves physical and mental health in women with fibromyalgia: a randomized controlled trial. AB - OBJECTIVE: To evaluate the feasibility of 8 months of supervised exercise therapy in warm water and its effects on the impact of fibromyalgia on physical and mental health and physical fitness in affected women. METHODS: Thirty women with fibromyalgia were randomly assigned to an exercise therapy group (n = 15) or a control group (inactive) (n = 15). The impact of fibromyalgia on physical and mental health was assessed using the Fibromyalgia Impact Questionnaire and the anxiety state with State-Trait Anxiety Inventory. Physical fitness was measured using the following tests: Canadian Aerobic Fitness; hand-grip dynamometry; 10 metre walking; 10-step stair-climbing and blind 1-leg stance. RESULTS: After 8 months of training, the exercise therapy group improved compared with the control group in terms of physical function (20%), pain (8%), stiffness (53%), anxiety (41%), depression (27%), Fibromyalgia Impact Questionnaire total scores (18%), State-Trait Anxiety Inventory score (22%), aerobic capacity (22%), balance (30%), functional capacity for walking (6%), stair-climbing with no extra weight (14%) and stair-climbing 10 kg-weighted (25%). CONCLUSION: Eight months of supervised exercise in warm water was feasible and led to long-term improvements in physical and mental health in patients with fibromyalgia at a similar magnitude to those of shorter therapy programmes. PMID- 18382820 TI - Impact of home modification services on ability in everyday life for people ageing with disabilities. AB - OBJECTIVE: To examine the impact of home modifications on self-rated ability in everyday life from various aspects for people ageing with disabilities. METHODS: The study sample was recruited from an agency providing home modification services in Sweden and comprised 73 subjects whose referrals had been approved and who were scheduled to receive home modifications (intervention group) and 41 subjects waiting for their applications to be assessed for approval (comparison group). The subjects rated their ability in everyday life using the Client Clinician Assessment Protocol Part I on 2 occasions: at baseline and follow-up. The Client-Clinician Assessment Protocol Part I provides data on the clients' self-rated independence, difficulty and safety in everyday life. The data were first subjected to Rasch analysis in order to convert the raw scores into interval measures. Further analyses to investigate changes in self-rated ability were conducted with parametric statistics. RESULTS: Subjects who had received home modifications reported a statistically significant improvement in their self rated ability in everyday life compared with those in the comparison group. Subjects who had received home modifications reported less difficulty and increased safety, especially in tasks related to self-care in the bathroom and transfers, such as getting in and out of the home. CONCLUSION: Home modifications have a positive impact on self-rated ability in everyday life, especially on decreasing the level of difficulty and increasing safety. PMID- 18382821 TI - Predicting the discharge destination of rehabilitation patients using a signal detection approach. AB - OBJECTIVE: To predict the discharge destination of rehabilitation patients using signal detection analysis. DESIGN: Cross-sectional and follow-up studies. SUBJECT: The subjects were 324 patients discharged from a hospital in Fukuoka, Japan, between April 2005 and March 2006 and 313 patients discharged from the same hospital between 1 April and 31 October 2006. METHODS: The discharge destinations of the 324 patients were predicted using signal detection analysis. As a validation study, 7 variables identified in the first analysis were used to categorize 313 patients, organized retrospectively into 8 groups, and to calculate the home discharge rate in each group. RESULTS: A patient's activities with respect to daily living, key person preference, dementia, age, route taken to hospitalization, residence before hospitalization, and gender were significant predictors of his or her discharge destination. Signal detection analysis established 8 subgroups, with 17.9-99.1% of the patients returning home after discharge. As a validation study, the actual and expected rates in the 8 subgroups were compared, and no significant difference was observed between the rates in any subgroup. CONCLUSION: Signal detection analysis is a useful technique for predicting the discharge destination of rehabilitation patients. PMID- 18382822 TI - Time-courses of lung function and respiratory muscle pressure generating capacity after spinal cord injury: a prospective cohort study. AB - OBJECTIVE: To investigate the time-courses of lung function and respiratory muscle pressure generating capacity after spinal cord injury. DESIGN: Multi centre, prospective cohort study. SUBJECTS: One hundred and nine subjects with recent, motor complete spinal cord injury. METHODS: Lung function and respiratory muscle pressure generating capacity were measured at first mobilization, at discharge from inpatient rehabilitation and one year after discharge. Lung function was measured in all 109 subjects, and 55 of these performed additional measurements of respiratory muscle pressure generating capacity. Trajectories of respiratory muscle function for different lesion level groups were assessed by multi-variate multi-level regression models. RESULTS: Forced vital capacity, forced expiratory volume in 1 sec and maximal inspiratory muscle pressure generating capacity significantly increased during and after inpatient rehabilitation. Forced inspiratory volume in 1 sec, peak inspiratory flow, peak expiratory flow and maximal expiratory muscle pressure generating capacity increased only during inpatient rehabilitation, but not thereafter. Increasing lesion level had a negative effect on all measured lung function parameters, as well as on maximal inspiratory and expiratory muscle pressure generating capacity. CONCLUSION: Respiratory function improved during inpatient rehabilitation, but only forced vital capacity, forced expiratory volume in 1 sec and maximal inspiratory muscle pressure generating capacity further improved thereafter. In particular, expiratory muscle function and subjects with tetraplegia should be screened and trained regularly. PMID- 18382823 TI - Shoulder pain in persons with thoracic spinal cord injury: prevalence and characteristics. AB - OBJECTIVE: To document the prevalence of shoulder pain, associated patient characteristics, and intensity and interference with shoulder function in activities in wheelchair users. DESIGN: A cross-sectional survey. PATIENTS: Individuals with a thoracic spinal cord injury. METHODS: A 3-part questionnaire on patient characteristics, medical history, supplemental questions, and the Wheelchair Users Shoulder Pain Index was administered to 101 patients. RESULTS: Of the 88 subjects (87%) who completed the questionnaire, 40% reported experiencing current shoulder pain, often chronic in nature. The highest median intensity of shoulder pain was reported for pushing the wheelchair up ramps or inclines outdoors. Reports of limitations regarding the ability to use the shoulder in activities were common. Most subjects used no upper extremity assistive devices during transfer or wheelchair propulsion. Of those with current shoulder pain, 37% had not sought medical attention. Age was associated with ongoing shoulder pain. CONCLUSION: Shoulder pain is a common problem in wheelchair users with a thoracic spinal cord injury, it seems more likely to occur with increased age, and is a potential cause of activity limitations. The findings indicate a need to develop programmes of prevention, patient education, diagnosis and treatment, possibly including more frequent use of upper extremity assistive devices. PMID- 18382824 TI - Determinants of improvement in walking capacity among individuals with chronic stroke following a multi-dimensional exercise program. AB - OBJECTIVE: To identify the determinants of improvement in walking capacity following therapeutic exercise in chronic stroke survivors. DESIGN: A secondary analysis of data obtained from a prospective, single-blind, randomized controlled intervention trial. SUBJECTS: Sixty-three community-dwelling individuals (mean age = 65 years, age range = 50-87 years) with a chronic stroke (post-stroke duration: mean = 5.5 years, range = 1-28 years). METHODS: Subjects were randomized into a leg exercise group (n = 32) or an arm exercise group (n = 31). Subjects in each group underwent 3 1-hour exercise sessions per week for 19 weeks. Walking capacity, cardiorespiratory fitness, isometric knee extensor muscle strength, balance ability, and balance confidence were evaluated before and after the interventions. Multiple regression analysis was performed to identify the determinants of improvement in walking capacity. RESULTS: After controlling for age, gender, post-stroke duration, and baseline walking capacity, gain in paretic leg muscle strength and peak oxygen consumption remained independently associated with gain in walking capacity (R2 = 0.229). CONCLUSION: Enhancement of cardiorespiratory fitness and paretic leg muscle strength are both significant determinants in improving walking capacity among chronic stroke survivors. However, the rather weak relationship (R2 = 0.229) indicates that other factors not measured in this study may also contribute to the improvement in walking capacity. PMID- 18382825 TI - A national survey of occupational therapists' practices related to participation post-stroke. AB - OBJECTIVES: First, to identify occupational therapists' stroke rehabilitation practices related to leisure and social aspects of participation and potential explanatory variables associated with these practices. Secondly, to identify occupational therapists' desired assessment and treatment practices related to participation. DESIGN: A Canada-wide telephone survey. SUBJECTS: A random sample of 480 occupational therapists providing stroke rehabilitation. METHODS: Two case studies were created: one representing a patient receiving inpatient stroke rehabilitation; the other receiving community-based rehabilitation. A standardized questionnaire was used to elicit information on: (i) clinician and environmental variables; (ii) management of the patient depicted; (iii) desired assessment and intervention use. RESULTS: 60.2% identified a problem relating to leisure or social aspects of participation, 23.1% would use an assessment and 36.5% would offer an intervention focusing on leisure or social participation. Desired assessment use was low (1%), as was desired intervention use (15.2%). Regression analyses using numerous potential explanatory variables explained little regarding clinician practices. CONCLUSION: Less than half of the occupational therapists focused interventions on leisure and social aspects of participation, suggesting a gap between what could be done to enhance successful community reintegration post-stroke and what is currently done. PMID- 18382826 TI - Inhibition of the unaffected motor cortex by 1 Hz repetitive transcranical magnetic stimulation enhances motor performance and training effect of the paretic hand in patients with chronic stroke. AB - OBJECTIVE: Recent reports demonstrated that low-frequency repetitive transcranial magnetic stimulation (rTMS) over the unaffected hemisphere improved the affected hand function in chronic stroke patients. We investigated whether 1 Hz rTMS improved the motor learning of the affected hand in patients after stroke. DESIGN: A double-blind study. PATIENTS: Twenty patients with chronic subcortical stroke. METHODS: The patients were randomly assigned to receive either a sub threshold rTMS over the unaffected hemisphere (1 Hz, 25 minutes) or sham stimulation, and all patients performed a pinching task after stimulation. We evaluated the motor function of the affected hand and the excitatory and inhibitory function of the affected motor cortex by transcranial magnetic stimulation. RESULTS: Compared with sham stimulation, rTMS induced an increase in the excitability of the affected motor cortex(p < 0.001) and an improvement in acceleration of the affected hand (p = 0.006). Moreover, the effect of motor training on pinch force was enhanced by rTMS (p < 0.001). These improvement in the motor function lasted for one week after rTMS and motor training (p < 0.001). CONCLUSION: rTMS improved the motor learning of the affected hand in patients after stroke; thus, it can apply as anew rehabilitation strategy for patients after stroke. PMID- 18382827 TI - Association between muscle function and low back pain in relation to pregnancy. AB - OBJECTIVE: To investigate the association of muscle function and subgroups of low back pain (no low back pain, pelvic girdle pain, lumbar pain and combined pelvic girdle pain and lumbar pain) in relation to pregnancy. DESIGN: Prospective cohort study. SUBJECTS: Consecutively enrolled pregnant women seen in gestational weeks 12-18 (n = 301) and 3 months postpartum (n = 262). METHODS: Classification into subgroups by means of mechanical assessment of the lumbar spine, pelvic pain provocation tests, standard history and a pain drawing. Trunk muscle endurance, hip muscle strength (dynamometer) and gait speed were investigated. RESULTS: In pregnancy 116 women had no low back pain, 33% (n = 99) had pelvic girdle pain, 11% (n = 32) had lumbar pain and 18% (n = 54) had combined pelvic girdle pain and lumbar pain. The prevalence of pelvic girdle pain/combined pelvic girdle pain and lumbar pain decreased postpartum, whereas the prevalence of lumbar pain remained stable. Women with pelvic girdle pain and/or combined pelvic girdle pain and lumbar pain had lower values for trunk muscle endurance, hip extension and gait speed as compared to women without low back pain in pregnancy and postpartum (p < 0.001-0.04). Women with pelvic girdle pain throughout the study had lower values of back flexor endurance compared with women without low back pain. CONCLUSION: Muscle dysfunction was associated with pelvic girdle pain, which should be taken into consideration when developing treatment strategies and preventive measures. PMID- 18382828 TI - Mirror therapy in patients with causalgia (complex regional pain syndrome type II) following peripheral nerve injury: two cases. AB - OBJECTIVE: To describe the use of mirror therapy in 2 patients with complex regional pain syndrome type II following traumatic nerve injury. DESIGN: Two case reports. SUBJECTS: Two patients with complex regional pain syndrome type II. METHODS: Two patients received mirror therapy with the painful hand hidden behind the mirror while the non-painful hand was positioned so that, from the perspective of the patient, the reflection of this hand was "superimposed" on the painful hand. Pain was measured with a visual analogue scale. RESULTS: The first case had developed a severe burning and constant pain in the hand due to a neuroma. In this patient, a strong reduction in pain was found during and immediately after mirror therapy. As a result, the patient was able to perform active exercises that were previously too painful. However, despite the pain relief during and directly after the exercises, the overall level of pain did not decrease. The second patient also had severe burning pain following a glass injury. In this patient, repeated mirror therapy for a 3-month period strongly decreased pain due to causalgia. CONCLUSION: The presented cases demonstrate that the use of mirror therapy in patients with causalgia related to a neuroma is worthy of further exploration as a potential treatment modality in patients with causalgia. PMID- 18382829 TI - Botulinum toxin type A in the healing of ulcer following oro-mandibular dyskinesia in a patient in a vegetative state. AB - OBJECTIVE: Use of botulinum toxin is expanding as clinical studies demonstrate new potential therapeutic applications. In rehabilitation, botulinum toxin is predominantly used as adjunct therapy for the treatment of spasticity, but it may prove useful for other atypical clinical situations. CASE HISTORY: A 73-year-old man had a severe sub-arachnoid haemorrhage following the rupture of a giant aneurism of the middle left cerebral artery. Clinically, the patient presented a vegetative state and an oro-mandibular dyskinesia that produced a chronic ulcer on the lower lip. As treatment for this dyskinesia, a total of 320 U botulinum toxin type A were injected into the upper and lower orbicularis oris and masseter muscles. RESULTS AND DISCUSSION: This treatment allowed for application of topical medication and subsequently, ulcer healing. Botulinum toxin type A may be an important therapeutic aid for clinicians faced with treating persistent pathological conditions caused by dyskinesia. PMID- 18382830 TI - The need for exercise rehabilitation in Botswana. PMID- 18382831 TI - Country-centered family medicine. PMID- 18382832 TI - Understanding the role of shame in the clinical assessment of health literacy. PMID- 18382833 TI - An audit of choosing and using the NHS Walk-in Centre in Skelmersdale, Lancashire, UK. PMID- 18382834 TI - Assembling patient-centered medical homes in teaching practices--one strategy. PMID- 18382835 TI - Life in 55 words: part I. PMID- 18382836 TI - Mistakes and disclosure. PMID- 18382837 TI - Which medical interview behaviors are associated with patient satisfaction? AB - OBJECTIVE: The objective of this study was to investigate the association between several medical interview behaviors and patient satisfaction. METHODS: The subjects were 158 new patients who visited an outpatient facility of a university hospital in Japan. All medical interviews were videotaped and reviewed by a trained rater using a medical interview rating scale (Takemura Medical Interview Rating Scale) for evaluating medical interview behaviors. To measure patient satisfaction, a self-administered questionnaire was also developed. Both the rating scale and the questionnaire were assessed for validity and reliability beforehand. RESULTS: A significant positive association was found between the behaviors of reflection and legitimation on the one hand, and patient satisfaction on the other. The positive association between reflection and patient satisfaction existed after adjusting for both the duration of the interview and the other medical interview behaviors used. The association between legitimation and patient satisfaction also existed after adjusting for the duration of the medical interview but disappeared after adjusting for the other medical interview behaviors used. When we investigated the strength of the relationship between each medical interview behavior and patient satisfaction, reflection was found to be the strongest determinant of patient satisfaction. CONCLUSIONS: This research revealed a significant positive association between reflection or legitimation and patient satisfaction in an actual clinical practice setting. PMID- 18382838 TI - Toward measuring the domains of mentoring. AB - BACKGROUND AND OBJECTIVES: Mentoring benefits faculty members and institutions, but mentoring efforts are difficult to sustain. Measuring the domains of mentoring could focus efforts to achieve desired outcomes. This project investigated the factor structure of a "mentoring experiences" measurement instrument. METHODS: A previously developed instrument was revised and distributed to 108 faculty members in one medical school department as part of the department's professional development process. RESULTS: The response rate was 89%. The instrument's Cronbach's alpha was 0.96, and factor analysis produced four factors with eigenvalues greater than 1 and factor loadings >0.400 for 27 items. DISCUSSION: This study determined that an instrument measuring domains of the protege's experience is reliable. This instrument may be useful to guide and evaluate mentoring initiatives. PMID- 18382839 TI - Preliminary study of a school-based program to improve hypertension awareness in the community. AB - BACKGROUND AND OBJECTIVES: We developed a school-based program to raise community awareness about hypertension. We studied the results of the program on the outcomes of parental knowledge about high blood pressure (BP) and their reports of having seen or intending to see a health professional about their BP. METHODS: We pilot tested a prototype middle-school curriculum using a crossover design in three fifth-grade classrooms (two designated early and one designated delayed intervention) in a suburban public school. We then used McNemar's test to assess significant differences in proportions responding correctly to BP knowledge questions and proportions reporting having seen or intending to see a health care professional about BP before and after the intervention. We tested for differences at 2-month follow-up in the early intervention classrooms compared to the delayed intervention classroom using chi-square. RESULTS: Seventy-six parents (out of a potential 134) completed baseline questionnaires. Parents had high baseline knowledge about certain aspects of hypertension, but baseline knowledge that high BP could lead to kidney failure was relatively low. The percentage of parents responding correctly to a question of whether high BP could lead to kidney failure increased after the intervention from 45.5% to 64.3%. Among parents in the early intervention classrooms, the percentage who reported having seen or intending to see a health professional about their BP increased from 9.6% to 27.5%. At 2-month follow-up, 27.5% of parents in the early intervention classrooms reported being seen or intending to be seen by a health care provider about their BP compared to only 8.3% of the parents in the delayed intervention classroom. CONCLUSIONS: This program may improve parents' knowledge about hypertension and their intent to be seen about BP. PMID- 18382840 TI - Training family physicians in community health centers: a health workforce solution. AB - PURPOSE: For more than 25 years, family medicine residencies (FMRs) have worked with community health centers (CHCs) to train family physicians. Despite the long history of this affiliation, little research has been done to understand the effects of training residents in this underserved community setting. This study compares CHC and non-CHC-trained family physicians regarding practice location, job and training satisfaction, and recruitment and retention to underserved areas. METHODS: We conducted a cross-sectional survey of a cohort of the 838 graduates from the WAMI (Washington, Alaska, Montana, and Idaho) Family Medicine Residency Network from 1986-2002. RESULTS: CHC-trained family physicians were almost twice as likely to work in underserved settings than their non-CHC-trained counterparts (64% versus 37%). When controlling for gender, percent full-time equivalent, and years from graduation, CHC-trained family physicians were 2.7 times more likely to work in underserved settings than non-CHC-trained family physicians. CHC and non-CHC-trained family physicians report similar job and training satisfaction and scope of practice. CONCLUSIONS: Training family physicians in CHCs meets the health workforce needs of the underserved, enhances the recruitment of family physicians to CHCs, and prepares family physicians similarly to their non-CHC trained counterparts. PMID- 18382841 TI - Revolutionary leadership and family medicine education. AB - Reform of the payment and delivery systems in American health care is now being discussed at the highest levels of business and government. Family medicine educators, researchers, and program leaders have an opportunity to provide substantial leadership to this process in their own communities and nationally. To do so, they must reconsider the assumptions made in creating our current systems of practice and education, and this will require new leadership skills that focus on innovation and adaptability. It will also require a more aggressive willingness to test new ideas and a new scientific method to prove or disprove their value. This essay outlines essential elements of such leadership for those responsible for the education of future generations of family physicians. PMID- 18382846 TI - Principles for the scope and practice of family medicine in Africa. PMID- 18382845 TI - Students' attitudes toward primary health care. PMID- 18382847 TI - A consensus of skills and procedures that residents need to learn in South Africa. PMID- 18382848 TI - First edition of the fourth volume of the International Journal of Prisoner Health. PMID- 18382849 TI - The right to health of prisoners in international human rights law. AB - This paper explores the health rights of prisoners as defined in international law, and the mechanisms that have been used to ensure the rights of persons in detention to realise the highest attainable standard of health. It examines this right as articulated within United Nations and regional human rights treaties, non-binding or so-called soft law instruments from international organisations and the jurisprudence of international human rights bodies. It explores the use of economic, social and cultural rights mechanisms, and those within civil and political rights, as they engage the right to health of prisoners, and identifies the minimum legal obligations of governments in order to remain compliant with human rights norms as defined within the international case law. In addressing these issues, this article adopts a holistic approach to the definition of the highest attainable standard of health. This includes a consideration of adequate standards of general medical care, including preventative health and mental health services. It also examines the question of environmental health, and those poor conditions of detention that may exacerbate health decline, disease transmission, mental illness or death. The paper examines the approach to prison health of the United Nations human rights system and its various monitoring bodies, as well as the regional human rights systems in Europe, Africa and the Americas. Based upon this analysis, the paper draws conclusions on the current fulfilment of the right to health of prisoners on an international scale, and proposes expanded mechanisms under the UN Convention against Torture and Other Cruel, Inhuman or Degrading Treatment to monitor and promote the health rights of prisoners at the international and domestic levels. PMID- 18382850 TI - Child sexual abuse, coping strategies and lifetime posttraumatic stress disorder among female inmates. AB - The purpose of the study was to assess the relationship between child sexual abuse (CSA) and posttraumatic stress disorder (PTSD) in female inmates in China. A total of 471 participants were assessed for CSA, personality traits, coping strategies, and frequency of lifetime PTSD. Results demonstrated that CSA and negative coping were predictive factors for lifetime PTSD among female inmates after personality traits were controlled. The findings suggest that CSA is one potential factor contributing to lifetime PTSD among female inmates. The importance of screening for CSA among female inmates was highly emphasized. Early intervention and prevention programmes based on coping skills may be useful to forestall the development of chronic PTSD in female inmates. PMID- 18382851 TI - Immunotoxicology IV - innovative methods and applications for risk assessment in pharmaceutical development: introduction to the conference. PMID- 18382852 TI - Hazard identification and risk assessment for biologics targeting the immune system. AB - Biologic pharmaceuticals include a variety of products, such as monoclonal antibodies, fusion proteins and cytokines. Products in those classes include immunomodulatory biologics, which are intended to enhance or diminish the activity of the immune system. Immunomodulatory biologics have been approved by the U.S. FDA for a variety of indications, including cancer and inflammatory conditions. Prior to gaining approval for marketing, sponsoring companies for all types of products must demonstrate a product's safety in toxicology studies conducted in animals and show safety and efficacy in clinical trials conducted in patients. The overall goal of toxicology studies, which applies to immunomodulatory and other product types, is to identify the hazards that products pose to humans. Because biologics are generally highly selective for specific targets (receptors/epitopes), conducting toxicology studies in animal models with the target is essential. Such animals are referred to as pharmacologically relevant. Endpoints routinely included in toxicology studies, such as hematology, organ weight and histopathology, can be used to assess the effect of a product on the structure of the immune system. Additionally, specialized endpoints, such as immunophenotyping and immune function tests, can be used to define effects of immunomodulatory products on the immune system. Following hazard identification, risks posed to patients are assessed and managed. Risks can be managed through clinical trial design and risk communication, a practice that applies to immunomodulatory and other product types. Examples of risk management in clinical trial design include establishing a safe starting dose, defining the appropriate patient population and establishing appropriate patient monitoring. Risk communication starts during clinical trials and continues after product approval. A combination of hazard identification, risk assessment and risk management allows for drug development to proceed with minimum risks to patients. PMID- 18382853 TI - Monoclonal antibody first dose cytokine release syndromes-mechanisms and prediction. AB - Acute cytokine release syndromes are associated with some therapeutic antibodies in man, leading to a spectrum of clinical signs from nausea, chills and fever to more serious dose limiting hypotension and tachycardia. When anticipated this syndrome is typically manageable, however this adverse reaction recently became headline news when a massive and unexpected cytokine release syndrome occurred within a few hours of dosing six healthy volunteers with a therapeutic antibody, putting their lives at risk due to multiple organ failure. Preclinical studies did not predict this adverse event, emphasising the need to compare the relative potency of the product in man and the chosen toxicology species, so that additional margins of safety can be applied when conducting first in man (FIM) studies if there is uncertainty over the predictability of the toxicology species. In vitro human PBMC and whole blood cultures may be useful for predicting cytokine release. However since cytokine release arises through at least two distinct mechanisms, it should be emphasised that the utility of these in vitro methods needs to be established for each antibody product. PMID- 18382854 TI - Urinary corticosterone as an indicator of stress-mediated immunological changes in rats. AB - Drugs that target the CNS or doses of drugs near the maximum tolerated dose may cause a non-specific stress response during routine safety testing in rodents that leads to the release of corticosterone and changes immunological parameters. In situations with mild clinical signs of stress and changes to immune organs, it may be difficult to differentiate direct immunotoxicity from changes mediated by stress. To address this concern, studies were conducted to identify potential biomarker of stress in rats that could be used in routine toxicology studies. Since serial blood collections for corticosterone levels are not practical, studies were conducted to evaluate urine corticosterone and its metabolites as a potential biomarker of stress in male Sprague-Dawley rats. Exogenous corticosterone was used as a reference to identify immune system targets and determine their relative sensitivity to corticosterone. The data from rats treated with exogenous corticosterone and from rats treated with drug or chemical stressors produced linear relationships between urine corticosterone and most immunological parameters, with r-squared values greater than 0.6. Thus, quantitatively similar effects on immunological end points are produced by exogenous corticosterone and by corticosterone induced by chemical stressors with regard to their correlation to selected immunological changes. In preclinical safety testing for a new drug, the combined findings of increased urinary corticosterone and changes of the predicted magnitude and direction in blood lymphocyte and neutrophil differentials and thymus weight or cellularity would strongly suggest that the immunological effects are secondary to a drug-induced stress response. Because these results can be obtained reliably during routine preclinical evaluations, they should be useful for the weight-of-evidence approaches often used in regulatory settings. PMID- 18382855 TI - Testing human biologicals in animal host resistance models. AB - The purpose of immunotoxicity testing is to obtain data that is meaningful for safety assessment. Host resistance assays are the best measure of a toxicant's effect on the overall ability to mount an effective immune response and protect the host from infectious disease. An outline is presented for immunotoxicological evaluation using host resistance assays. The influenza virus host resistance model is useful to evaluate the overall health of the immune system and is one of the most thoroughly characterized host resistance models. Viral clearance requires all aspects of the immune system to work together and is the ultimate measure of the health of the immune system in this model. Mechanistic immune functions may be included while measuring viral clearance and include: cytokines, macrophage activity, natural killer (NK) cell activity, cytotoxic T-lymphocyte (CTL) activity, and influenza-specific IgM and IgG. Measurement of these immunological functions provides an evaluation of innate immunity (macrophage or NK activity), an evaluation of cell-mediated immunity (CMI) (CTL activity), and an evaluation of humoral-mediated immunity (HMI) (influenza-specific IgM or IgG). Measurement of influenza-specific IgM or IgG also provides a measurement of T dependent antibody response (TDAR) since influenza is a T-dependent antigen. There are several targeted host resistance models that may be used to answer specific questions. Should a defect in neutrophil and/or macrophage function be suspected, Streptococcus pneumoniae, Pseudomonas aeruginosa, or Listeria monocytogenes host resistance models are useful. Anti-inflammatory pharmaceuticals or therapeutics for rheumatoid arthritis or Crohn's disease that target TNFalpha may also be evaluated for immunotoxicity using the S. pneumoniae intranasal host resistance assay. Marginal zone B (MZB) cells are required for production of antibody to T-independent antigens such as the polysaccharide capsule of the encapsulated bacteria that are so prominent in causing blood-borne infections and pneumonia. Intravenous infection with Streptococcus pneumoniae, an encapsulated bacterium, results in a blood-borne infection that requires MZB cells for clearance. The systemic S. pneumoniae host resistance assay evaluates whether a therapeutic test article exerts immunotoxicity on MZB cells and measures the T-independent antibody response (TIAR). Suppression of CMI or in some cases HMI may result in reactivation of latent virus that may result in a fatal disease such as progressive multifocal leukoencephalopathy (PML). The murine cytomegalovirus (MCMV) reactivation model may be used to evaluate a pharmaceutical agent to determine if suppression of CMI or HMI results in reactivation of latent virus. Candida albicans is another host resistance model to test potential immunotoxicity. Host resistance assays have been the ultimate measure of immunotoxicity testing for environmental chemicals and pharmaceutical small molecules. Human biologicals are now an important component of the drug development armamentarium for biotech and pharmaceutical companies. Many human biologicals are fusions of IgG, and/or target immune mediators, immunological receptors, adhesion molecules, and/or are indicated for diseases that have immune components. It is therefore necessary to thoroughly evaluate human biological therapeutics for immunotoxicity. Numerous biologicals that are pharmacologically active in rodents can be evaluated using well-characterized rodent host resistance assays. However, biologicals not active in rodents may use surrogate biologicals for testing in rodent host resistance assays, or may use host resistance assays in genetically engineered mice that mimic the effect of the human biological pharmacological agent. PMID- 18382856 TI - Adverse consequences of immunostimulation. AB - The therapeutic uses of immunostimulatory agents are generally in the treatments of infections or cancer. The traditional example of vaccination is one form of immunostimulation used in the prevention of pathogenic infections or cancer (e.g., human papillomavirus vaccine). Recombinant cytokines are increasingly used to stimulate immune system function. For example, interferon-alpha (IFNalpha) and interleukin (IL)-2 have been used to treat chronic hepatitis C virus infection and metastatic melanoma, respectively. In contrast, monoclonal antibodies are used to target malignant cells for elimination via antibody-dependent cytotoxicity mechanisms or apoptosis, including the anti-CD20 monoclonal antibody rituximab and the anti-CD56 monoclonal antibody alemtuzumab used in the treatment of B-cell malignancies, and the anti-erb2 receptor antibody trastuzumab used in the treatment of breast cancer. Finally, immunostimulation may develop via modulation of pathways involved in immune system regulation. For example, the anti-CD28 monoclonal antibody TGN1412 was developed as an agonist of regulatory T cells for treatment of T-cell-mediated chronic inflammatory diseases or leukemias. A panel was convened to discuss potential toxicities associated with immunostimulation. At the Immunotoxicology IV meeting in 2006, a panel, moderated by Dr. Robert House (Dynport Vaccine Co., Frederick, MD), included Drs. Gary Burleson (Burleson Research Technologies, Inc., Raleigh, NC), Kenneth Hastings (US FDA, Center for Drug Evaluation and Research [CDER], Rockville, MD), Barbara Mounho (Amgen, Thousand Oaks, CA), Rafael Ponce (ZymoGenetics, Inc., Seattle, WA), Mark Wing (Huntington Life Sciences, Cambridgeshire, United Kingdom), Lauren Black (Navigators Consulting, Sparks, NV) and Anne Pilaro (US FDA, CDER, Rockville, MD). This paper reviews the major identified toxicities associated with immunostimulation, including the acute phase response, cell and tissue abnormalities/injury, cytokine release/cytokine storm, tumor lysis syndrome, vascular leak, and autoimmunity that were discussed by this panel. PMID- 18382857 TI - IL-23/IL-17 biology and therapeutic considerations. AB - CD4+ T-lymphocytes are pivotal in immune responses to pathogens and in pathogenesis of various inflammatory diseases. Upon activation by the innate immune system, naive CD4+ helper T (T(H))-lymphocytes differentiate into effector cells that are characterized by their cytokine profiles. T-Helper 1 (T(H)1) and T(H)2 lymphocytes have long been regarded as two sides of a coin in adaptive immune responses. However, recent data indicate a novel lineage of T(H) lymphocytes specified by the production of interleukin-17 (IL-17). Here I describe the recent literature on the regulatory mechanisms underlying generation of IL-17-expressing T-lymphocytes. In addition, I also summarize on the biological function of these cells in infection, autoimmunity and allergy. IL-17 and IL-17-expressing inflammatory T(H) lymphocytes may thus serve as good targets for therapeutic intervention in treatment of patients with inflammatory diseases. PMID- 18382858 TI - Beta-glucans, history, and the present: immunomodulatory aspects and mechanisms of action. AB - The present paper represents a comprehensive up-to-date review of beta -glucans, their chemical and biological properties, and their role in immunological reactions. beta -D-Glucans belong to a group of physiologically active compounds called biological response modifiers and represent highly conserved structural components of cell walls in yeast, fungi, or seaweed. Despite almost 150 years of research, the exact mechanisms of their action remain unclear. The present review starts with the history of glucans. Next, attention is focused on sources and structure, comparing the effects of physicochemical properties, and sources on biological effects. As glucans belong to natural products useful in preventing various diseases, they have been highly sought after throughout human history. Based on extensive recent research, this paper explains the various mechanisms of effects and the ways glucans mediate their effects on defense reactions against infections. Despite the fact that predominately pharmacological effects of glucans are positive, their unfavorable and potentially toxic side effects were not overlooked. In addition, attention was focused on the future research, possible alternatives such as synthetic oligosaccharides, and on clinical applications. PMID- 18382859 TI - Pathogenic role of environmental toxins in immune-mediated liver diseases. AB - Immune-mediated liver diseases contribute significantly to morbidity and mortality due to liver failure and the need for liver transplantation. The pathogenesis of the immune-mediated chronic liver diseases, primary sclerosing cholangitis, autoimmune hepatitis, and primary biliary cirrhosis, is poorly understood. Genetic susceptibility factors may play a role, but increasing attention is being given to the association between environmental factors and these diseases. The existence of such a relationship is supported by epidemiologic surveys, animal models, and geographic clustering analyses. Unearthing the cause of this association may provide insight into the pathogenesis of immune-mediated chronic liver diseases and autoimmunity. PMID- 18382860 TI - Green tea attenuates benzene-induced oxidative stress in pump workers. AB - Workers exposed to benzene frequently suffer from toxicities of the bone marrow as well as the central nervous, immune, and reproductive systems. This toxicity most likely is a result of the oxidative metabolism of benzene to reactive products. As green tea possesses antioxidant effects, the objective of this study was to examine any amelioration of benzene-induced oxidative stress in pump workers drinking 6 cups (150 ml/cup) of freshly prepared tea daily. Sixty male non-smoking subjects, divided into four groups: no benzene exposure/no green tea; no exposure/tea; exposure/no tea; and, exposure/tea, were monitored after a 6 mo period. On the final day of the study, urine samples were collected for analyses of benzene, trans-trans muconic acid, and phenol. Blood was also collected at this time; plasma was assayed for total antioxidant activity, malondialdehyde (MDA), and glutathione (GSH) while erythrocytes were analyzed for activity of antioxidant enzymes glutathione peroxidase (GSHPX), superoxide dismutase (SOD), and catalase. The results demonstrated that urinary levels of benzene, trans trans muconic acid, and phenol were elevated in all pump workers, and that this elevation was mitigated by consumption of green tea. The benzene exposures also led to significant reductions in plasma GSH levels and erythrocyte antioxidant enzyme activities; these effects were abrogated (to near-control levels) by the tea. Interestingly, among control subjects, tea ingestion itself caused significant increases in both GSHPX and catalase activities. Unlike with the other plasma parameters, while the benzene exposures also significantly increased plasma MDA levels and decreased total antioxidant activity, tea ingestion did not cause a near-total reversion to control values; the effects on these two endpoints were more like those noted with the urine parameters (mitigation, not abrogation). These studies demonstrate that drinking green tea during benzene exposure can reduce several parameters indicative of oxidative stress. As such, as a dietary supplement, green tea could represent a potential therapeutic agent in reducing certain aspects of benzene-induced toxicity. PMID- 18382861 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin alters the differentiation of alloreactive CD8+ T cells toward a regulatory T cell phenotype by a mechanism that is dependent on aryl hydrocarbon receptor in CD4+ T cells. AB - Activation of aryl hydrocarbon receptor (AhR) by 2,3,7,8-tetracholordibenzo- p dioxin (TCDD) during an acute graft-versus-host response induces a population of alloreactive donor CD4+CD25+ regulatory T (Treg)-like cells that have potent suppressive activity in vitro. In the present studies, we show that TCDD induced a similar population of donor CD8+CD25+ T-cells with suppressive activity in vitro. Like the CD4+ Treg cells, donor CD8+CD25+ T-cells also expressed higher levels of CD28, glucocorticoid-induced TNFR (GITR) and CTLA-4 along with low levels of CD62L. These TCDD-induced phenotypic changes were not observed if donor T-cells were obtained from AhR-KO mice. When CD4+ and CD8+ donor T-cells from AhR WT and AhR-KO mice were injected in various combinations into F1 mice, the enhanced expression of CD25 on CD8+ T-cells required AhR in donor CD4+ T-cells, while down-regulation of CD62L required AhR in the donor CD8+ T-cells themselves. Changes in GITR and CTLA-4 on donor CD8+ T-cells were partially mediated by AhR in both T-cells subsets. In contrast, all phenotypic changes in donor CD4+ T cells were dependent on the presence of AhR in the CD4+ T-cells themselves. These findings suggest that the direct effects of AhR-mediated signaling in CD8+ T cells are more limited than the direct effects in CD4+ T-cells, and that AhR signaling in CD4+ T-cells may be a unique pathway for the induction of both CD4+ and CD8+ adaptive Treg. PMID- 18382862 TI - Womens' Day and some valid health issues for women. PMID- 18382863 TI - Pelvic floor muscle training in the prevention and treatment of urinary incontinence in women - what is the evidence? AB - Many women suffer from urinary incontinence (UI). During and after pregnancy, women are advised to perform pelvic floor muscle training (PFMT) to prevent the development of UI. In established UI, PFMT is prescribed routinely as first-line treatment. Published studies are small, underpowered and of uneven methodological quality. Variations in study populations, intervention types and outcome measures make comparisons difficult. While further studies are needed, the available evidence suggests a lack of long-term efficacy of peripartum PFMT. In established UI, there seems to be a modest immediate response to PFMT. Based on the available evidence, we believe that a critical reappraisal of PFMT is needed, and judgments on the place of PFMT in current clinical practice should be reserved until further evidence, including cost-benefit analyses, has unequivocally demonstrated a clinically relevant efficacy. PMID- 18382864 TI - Placental pathology in women with gestational diabetes. AB - BACKGROUND: The aim of this study was to investigate pathologic differences of the placenta in pregnancies complicated by gestational diabetes compared to non diabetic pregnancies. METHODS: Forty singleton pregnancies complicated by gestational diabetes were recruited and compared to 40 consecutive normal pregnancies. A pathologist, blinded to all clinical data, reviewed all histological samples of the placentas. The histological assessment was carried out with regard to the following aspects: fetal vessel thrombosis, villous immaturity, chorangiosis, presence of nucleated fetal red blood cells (NFRBCs), ischemia, infarction, presence of hydropic or avascular villi, lymphohistiocytic villitis and villous fibrinoid necrosis. RESULTS: The presence of degenerative lesions such as fibrinoid necrosis and vascular lesions like chorangiosis was apparent, mainly in the diabetes group. Villous immaturity and the presence of NFRBC as an indication of chronic fetal hypoxia were significantly increased in the placentas of women with diabetes compared with the control group. Fetal/placental weight ratio was significantly lower in the diabetic group. CONCLUSION: Histological abnormalities were observed more frequently in the diabetic placentas compared to the controls. These findings support the hypothesis that impaired placental function is one of the main reasons for the increased frequency of fetal complications in diabetic pregnancies. PMID- 18382865 TI - Impaired maternal cognitive functioning after pregnancies complicated by severe pre-eclampsia: a pilot case-control study. AB - BACKGROUND: Pre-eclampsia is the most significant cause of neurological symptoms in pregnancy. Neurological symptoms may persist even after pregnancy. Somatic symptoms of pre-eclampsia, such as hypertension and proteinuria, generally disappear after delivery. However, formerly pre-eclamptic women more often complain of cognitive disturbances compared to women after uncomplicated pregnancies. METHODS: Three to eight months postpartum, a neuropsychological test battery was performed in 10 former severely pre-eclamptic women (according to the guidelines of the American College Obstetricians and Gynecologists) and 10 women after uncomplicated normotensive pregnancies. The control group was matched for age, educational level and mode of anesthesia. All women delivered by cesarean section either under general or regional anesthesia. Tests were performed for premorbid intelligence, short- and long-term memory, attention, concentration, executive functions, visual and spatial abilities. Anxiety and depression levels were measured. RESULTS: The formerly pre-eclamptic women had significantly lower scores on most indices of the auditory-verbal memory test. Formerly pre-eclamptic patients learned considerably fewer words than controls and recalled less after interference. Both case and control group did not differ in age, parity or level of education. There were no differences in the level of intellectual functioning and language tests, such as naming and word fluency. No persistent differences were observed in tests for attention/concentration and executive functioning. There were no significant differences on depression and anxiety scales. CONCLUSIONS: Maternal memory seems to be impaired after pregnancies complicated by severe pre-eclampsia. This effect cannot be attributed to depression and/or anxiety or method of anesthesia. PMID- 18382866 TI - Serum visfatin levels in late pregnancy and pre-eclampsia. AB - OBJECTIVES: To characterise the changes in serum visfatin levels in late normal pregnancy and pre-eclampsia. METHODS: Twenty-seven women with pre-eclampsia were recruited. Twenty-eight women in the third trimester of normal pregnancy served as pregnant control and 28 healthy non-pregnant women as non-pregnant control. Serum levels of visfatin were measured with an enzyme-linked immunosorbent assay. RESULTS: The means of serum visfatin were 626.4+/-45.5 ng/ml (mean+/-SEM) in non pregnant control, 695.9+/-92.5 ng/ml in pregnant control, and 308.3+/-80.0 ng/ml in pre-eclampsia, respectively, and were significantly different among the groups (p<0.001). Visfatin level was significantly lower in pre-eclampsia compared to non-pregnant control (p=0.004) and pregnant control (p<0.001). Women with severe pre-eclampsia had a significantly lower serum visfatin level than those with mild pre-eclampsia (114.6+/-80.9 versus 425.2+/-122.1 ng/ml, p=0.037). CONCLUSIONS: A decrease in visfatin level was demonstrated in pre-eclampsia, suggesting that visfatin and adipokine-associated metabolic abnormalities are involved in the pathogenesis of the disease. PMID- 18382867 TI - Pregnancy outcomes of women with pruritus, normal bile salts and liver enzymes: a case control study. AB - BACKGROUND: Obstetric cholestasis (OC) is associated with increased maternal and perinatal complications. Nevertheless, data on pregnancy outcomes of women who experience pruritus on a transient basis, but have normal bile salts and liver function tests (LFT) is scarce. METHODS: The maternal and fetal outcomes of 144 women with pruritus but normal bile salts and LFTs were compared with the next delivered patient without itch who matched for age, ethnicity and parity. RESULTS: The study and control groups had similar mean gestational ages at delivery and birth weights (p>0.05, t test). However, women with pruritus were more likely to have meconium-stained liqor, abnormal intrapartum cardiotocography and postpartum hemorrhage (PPH) (p<0.05, Fisher's exact test). There appears to be a trend towards a higher rate of instrumental delivery (p=0.07) in the study compared to the control group, although this did not reach statistical significance. CONCLUSION: This study suggests that women who have transient pruritus with normal bile salts and liver biochemistry appear to have higher intrapartum and postpartum complications and require increased vigilance. In order to evaluate this finding, further prospective studies will be required. PMID- 18382868 TI - Modifiable determinants of fetal macrosomia: role of lifestyle-related factors. AB - BACKGROUND: Newborn macrosomia is associated with both short- and long-term health risks for the infant, and increases the prevalence of birth complications. Parity, maternal age and gender of the child are known variables that influence fetal growth. The purpose of the present investigation was to evaluate prospectively the contributions of modifiable maternal predictors of fetal macrosomia (> or =4,200 g), which included lifestyle-related factors, such as nutritional intake, physical activity, and plasma glucose values, in addition to overweight and pregnancy weight gain. METHODS: Some 553 women were followed through pregnancy. Predictive variables were subjected to univariate and multiple logistic regression analysis. Among these were: body mass index (BMI), weight gain, maternal subcutaneous fat (mm), fasting and 2-h plasma glucose, self reported physical activity before and during pregnancy, and nutritional intake of macronutrients. Gestational age, parity and gender were also included in the model. All continuous variables were dichotomized using the upper quartile as the cut-off point in most cases. RESULTS: If physical activity was left out of the analyses, BMI, weight gain, plasma glucose and gestational age were independent determinants of macrosomia. After including low level pre-gestational physical activity in the model, we found that this was now a significant determinant of delivering a macrosomic infant with an OR=2.9 (95% CI: 1.9, 7.3). CONCLUSION: The present study indicates that a low level of pre-gestational physical activity adds to the modifiable determinants of newborn macrosomia. PMID- 18382869 TI - Interleukin-10 -1082 G/A promoter polymorphism and pregnancy complications: results of a prospective cohort study in 1,616 pregnant women. AB - OBJECTIVE: To investigate the frequency of the interleukin-10 (IL-10)-1082 G/A single nucleotide polymorphism in women with intrauterine fetal death (IUFD), pre eclampsia (PE), preterm delivery (PD), and small for gestational age (SGA) infants. METHODS: In a prospective cohort study, DNA from 1,616 consecutive pregnant women was analyzed for IL-10 -1082 G/A by polymerase chain reaction. Women who developed at least one of the predefined pregnancy complications were used as cases and compared to women without pregnancy complications. RESULTS: Of 1,616 women, 254 (15.7%) developed at least one pregnancy complication. IL-10 1082 G/A allele frequencies (G: 233/508 [45.9%] and A: 275/508 [54.1%] versus G: 1,143/2,724 [42.0%] and A: 1,581/2,724 [58.0%], respectively; p=0.10; OR 0.85; 95% CI 0.69-1.04) and genotype distributions (A/A+G/A: 201/254 [79.1%] and G/G 53/254 [20.9%] versus A/A+G/A: 1,125/1,362 [82.6%] and G/G 237/1,362 [17.4%], respectively, p=0.19; OR 0.79; 95% CI 0.54-1.15) were not significantly different between cases and controls. We observed no statistically significant difference in IL-10 -1082 G/A genotype distribution comparing controls and women with IUFD, PE, PD <37 weeks gestation, and SGA infants (<10th percentile). CONCLUSION: IL-10 -1082 G/A polymorphism is not a genetic marker for identifying women at increased risk of common pregnancy complications. PMID- 18382870 TI - Employing nifedipine as a tocolytic agent prior to external cephalic version. AB - BACKGROUND: External cephalic version (ECV) is accepted as a means of reducing the rate of breech presentation at term. Routine use of tocolysis prior to an ECV has been described to increase its success rate. The study's objective was to evaluate the efficacy of oral nifedipine as a tocolytic agent prior to ECV, and to compare it with intravenous ritodrine (IR). METHODS: Women with breech presentation at term considered suitable for ECV were given nifedipine prior to the procedure. The success rate was compared to a cohort of women who underwent an ECV attempt at our department during the years 1999-2002 using IR prior to the procedure. Power analysis indicated that 70 women were needed in each group to detect a difference of 25% from our baseline success rate of 50%. RESULTS: Seventy-six women receiving nifedipine before their ECV attempt were compared to 90 women who received IR. Age, parity, gestational age at ECV, type of breech, placental location, fetal weight, and gestational age at delivery were not different between the groups. The overall success rate was 54% and 50% in the nifedipine and ritodrine groups respectively (p=0.6). The success rate among primiparous women was 35% and 29% respectively (p=0.8). The success rate among multiparous women was 64% and 63% respectively (p=1.0). The success rate was statistically different between primiparous and multiparous women in each group (p<0.05). CONCLUSION: Oral nifedipine may be as effective as IR when administered before ECV. PMID- 18382871 TI - Attitudes towards cesarean section in a nationwide sample of obstetricians and gynecologists. AB - OBJECTIVE: The Swedish cesarean section (CS) rate has increased dramatically over the last decades, but remains relatively low compared to several other countries in the Western world. The reasons for this continuous increase are multifactorial and difficult to investigate. That pregnant women are demanding CS more frequently does not appear to be the major explanatory factor for the increasing CS rate. Since Swedish CS rates are relatively low, it is important to study attitudes to CS in a nationwide population of Swedish obstetricians and gynecologists in order to determine the possible concerns of this group. METHODS: In 2006, a study-specific questionnaire was created and posted to Swedish obstetricians and gynecologists. In total, 1,280 obstetricians and gynecologists received a questionnaire and replied with a response rate of 66%. RESULTS: In general, older and longer work experience physicians had a more positive attitude towards providing CS on maternal request or elective CS for women who fear vaginal delivery. In addition, these colleagues were more positive in expressing their belief that elective CS is as safe as vaginal delivery for the mother and her child. The main difference between female and male physicians was that males were more positive toward providing CS on maternal request (p<0.001). CONCLUSION: We have shown that female physicians differ from their male colleagues in their attitudes toward providing CS on maternal request. Female physicians were more reluctant than males to provide CS on maternal request. PMID- 18382872 TI - Transrectal ultrasound and the diagnosis of rectovaginal endometriosis: a prospective observational study. AB - BACKGROUND: Rectovaginal endometriosis is a severe form of pelvic endometriosis in which pharmacological treatment is relatively ineffective (Vercellini et al., Fertil Steril. 2005;84:1375-87). Laparoscopic surgical treatment is effective, but has the potential risks of bowel perforation and colostomy formation (Darai et al., Am J Obstet Gynecol. 2005;192:394-400). Transrectal ultrasound scanning can be applied as a preoperative tool to predict the presence of rectovaginal endometriosis and bowel wall involvement (Abrao et al., J Am Assoc Gynecol Laparosc. 2004;11:50-4). METHODS: Thirty-two women underwent transrectal ultrasound followed by therapeutic laparoscopy. Likelihood ratios and post-test prevalences were calculated with Fagan's normogram. This was then extrapolated with the aid of a mathematical model to a low-risk population. RESULTS: A positive likelihood ratio was found to be 10.89 (95% confidence ratio (CI): 1.62 73.15) and a negative likelihood ratio was found to be 0.24 (95% CI: 0.1-0.57). The pre-test prevalence of rectovaginal endometriosis was 56%. The positive post test prevalence probability was 93%, and the negative post-test prevalence probability was 23%. CONCLUSION: Preoperative transrectal ultrasound scanning for rectovaginal endometriosis is an extremely accurate predictive test, and strongly predicts the need for extensive laparoscopic dissection and potential bowel resection. PMID- 18382873 TI - Genital prolapse surgery after a shift in treatment tradition: an analysis of subsequent prolapse surgery. AB - OBJECTIVE: To determine whether the shift in treatment tradition of genital prolapse surgery was followed by a change in the occurrence of subsequent prolapse surgery, and to analyze the complementary and recurrent surgery with respect to the size of the prolapse and the extent of the primary surgery. METHODS: A retrospective study of patients consecutively operated with primary prolapse surgery in three Swedish hospitals in two time periods: 261 patients in 1983 (Period I) and 281 patients in 1993 (Period II). Clinical data were obtained from the patient records. A follow-up period of 6 years was used for both periods. RESULTS: Subsequent prolapse surgery was seen significantly more often in Period II than in Period I (7.7 versus 2.7%), and after selective repairs compared with complete repairs (7.7 versus 3.2%). Despite a significant reduction in the use of posterior repair between the time periods, no significant increase was seen in complementary posterior repairs compared with complementary repair in any of the other compartments. Size of the prolapse at the primary surgery or hysterectomy did not seem to influence the occurrence of subsequent prolapse surgery. The postoperative complication rate was significantly higher after complete repairs than after selective repair, and especially when posterior repair was included in the operation. CONCLUSION: Subsequent prolapse surgery is slightly more common after selective repair than after complete repair. However, selective repairs are encumbered with a lower complication rate. The results of this study appear to be in favor of a restrictive use of 'prophylactic' posterior repair. PMID- 18382874 TI - MRI anatomical study of the outside-in transobturator suburethral tape procedure. AB - BACKGROUND: This study aims to clarify the relationship between the outside-in transobturator suburethral tape and the anatomical structures at the blind passage of the obturator foramen, and to detail the tape position, shape, and the factors on which they are dependent. METHODS: MRI scans were performed on 30 patients operated on with a dermal porcine collagen suburethral tape, indicated for urodynamically proven stress incontinence. RESULTS: The lateral bladder wall was the closest structure to the tape at the passage of the obturator foramen, whereas vascular injuries appeared to be minimal using the outside-in technique. The distance from tape to bladder neck was correlated with the distance from tape to pubococcygeal line (PCL) (p=0.001), and with urethral length (p=0.049). From the obturator foramen to its passage under the urethra, the tape was U-shaped with a cranio-caudal and antero-posterior orientation. The cranio-caudal tape shape was correlated with the distance between the tape and the PCL (p=0.002), and to the patient's anatomical characteristics: distance between the bladder neck and the pubis (p=0.01), and the urethral length (p=0.01). The antero posterior shape was correlated with the urethral length (p=0.02). CONCLUSION: The lateral bladder wall is the structure most at risk during the outside-in transobturator suburethral tape procedure. The distance from tape to bladder neck is dependent on the vertical position of the tape. The tape shape is dependent on both tape position under the urethra and the patient's anatomical characteristics. PMID- 18382875 TI - Anterior vaginal wall reconstruction: anterior colporrhaphy reinforced with tension free vaginal tape underneath bladder base. AB - OBJECTIVE: To assess the effectiveness of the polypropylene tape in preventing cystocele recurrence. METHODS: Fifty Caucasian women aged 50-77 years (mean age 66.6 years), with stage II-IV pelvic organ prolapse, enrolled into the study. Vaginal reconstructive surgery included an anterior colporrhaphy on all patients, posterior colpoperineorrhaphy on 28 patients and hysterectomy on 36 patients. Patients were divided into a study group of 28 women and a control group of 22 women. As reinforcement to the anterior colporrhaphy procedure, in the study group a polypropylene tape (TVT-O) was placed underneath the bladder base and fixed with polyglactin sutures. Postoperative follow-up was carried out every 4 months (total 48 months). The assessment of the anatomic result included both clinical evaluation of the operated sites and perineal sonography. RESULTS: The mean postoperative distance of the bladder base to the inferior edge of the symphysis pubis was 1.5 cm (range: 1.0-2.2 cm) in the study group and 2.8 cm (range: 2.0-3.8) in the control group. Postoperatively, there were two cases of stress incontinence and two cases of urge incontinence, one in each group. No case of tape erosion was noted, no dyspareunia and no recurrent cystocele in the study group. Four cases of recurrent cystocele (20%) were reported in the control group. CONCLUSION: While the preliminary results of our study are encouraging, larger series of patients and longer follow-up are required to verify the effectiveness of the aforementioned modification. PMID- 18382876 TI - Long-term consequences of gynecological cancer treatment on urinary incontinence: a population-based cross-sectional study. AB - OBJECTIVE: To study urinary incontinence among long-term survivors of gynecological cancer. STUDY DESIGN: In a population-based, cross-sectional design, we identified 319 recurrence-free survivors of gynecological cancer, and 1,276 controls without a history of gynecological cancer. All participants were addressed with a 16-pages questionnaire covering issues of quality of life, general health, daily living conditions, natural functions and co-morbidities. Following 1 reminder, the response rates were 55 and 41% for cases and controls, respectively. Urinary incontinence (total, stress, urge and mixed) was in agreement with definitions of the International Continence Society. RESULTS: Mean relapse-free follow-up time after treatment was 12 years (range: 8-17 years) for cases. The prevalence of total, stress, urge and mixed urinary incontinence were 34.3, 24, 0.8 and 9.5%, respectively. Previous gynecological cancer treatment was not associated with any outcomes of urinary incontinence. Obesity, previous and current use of HRT were associated with total, stress and mixed urinary incontinence, whereas increasing parity order was associated with total and stress urinary incontinence. In comparison with single factor analyses, the combination of obesity and parity 2+ had a multiplicative effect on total, stress and mixed incontinence. CONCLUSION: Recurrence-free long-term survivors of gynecological cancer are not at increased risk for urinary incontinence. PMID- 18382877 TI - Prognostic significance of the micropapillary pattern in patients with serous borderline ovarian tumors. AB - BACKGROUND: The impact of micropapillary histology on survival in patients with serous borderline ovarian tumors is not established. The purpose of this study was to evaluate the significance and influence of micropapillary pattern on clinical outcome of patients with serous borderline ovarian tumors. METHODS: Eighty-five patients who underwent surgery and were diagnosed with serous borderline ovarian tumors were retrospectively studied. Among these patients, 18 patients showed micropapillary pattern on histologic examination, while the remaining 67 patients did not. The significance of the presence of micropapillary pattern and relationship with various clinicopathologic findings and progression free survival were evaluated by statistical analysis. RESULTS: No differences with regard to age, parity, body mass index, pretreatment CA-125 level, tumor size, bilaterality, and the duration of follow-up were observed between the two groups, but there was a significant difference in terms of FIGO stage (p<0.001), invasive implants (p=0.004), performing lymphadenectomy (p=0.009), operation time (p=0.003), estimated blood loss (p<0.001), residual disease>1 cm (p=0.001), adjuvant chemotherapy (p<0.001), and the length of hospital stay (p=0.013). Progression-free survival was found to be significantly decreased in patients with invasive implants (HR, 5.06; p=0.025) and micropapillary pattern (HR, 4.20; p=0.056) on multivariate analysis. CONCLUSIONS: Micropapillary serous borderline ovarian tumors were associated with a more aggressive clinical course compared with typical serous borderline ovarian tumor. The presence of invasive implants and micropapillary pattern were significant prognostic factors in patients with serous borderline ovarian tumors. PMID- 18382879 TI - Sex selection, gender-based violence and human rights abuse. PMID- 18382882 TI - Introducing human papillomavirus vaccines - questions remain. AB - Genital human papillomavirus (HPV) infection and HPV-associated cervical and other anogenital cancers are significant public health problems. HPV 16 and HPV 18 are responsible for approximately 70% of all invasive cervical cancers worldwide. The first prophylactic HPV virus-like particle (VLP) vaccine against HPV types 6/11/16/18 was licensed in 2006 for girls and women aged 9-26 years. The second prophylactic HPV vaccine against HPV types 16 and 18 has been licensed this year. These vaccines are almost 100% effective in preventing infection and high-grade precancer associated with the HPV types included in the vaccine. The vaccines are well tolerated, safe, and highly immunogenic when given in three doses within 6 months. Efficacy of the vaccine against external vulvar and HPV related vaginal lesions is also high. Even though the vaccine is highly effective against high-grade cervical, vaginal, or vulvar precancers, this only applies to women unexposed to these HPV types and only to high-grade intraepithelial lesions caused by these HPV types. Therefore, it is important to understand that the population impact of the vaccines will be much lower than vaccinating naive populations. Implementing HPV vaccine is a great opportunity but also a great challenge. However, mandatory HPV vaccination may raise many questions, and more answers are needed. PMID- 18382883 TI - Genetic contributions to preterm birth: implications from epidemiological and genetic association studies. AB - Infants born before term (<37 weeks) have an increased risk of neonatal mortality as well as other health problems. The increasing rate of preterm birth in recent decades, despite improvements in health care, creates an impetus to better understand and prevent this disorder. Preterm birth likely depends on a number of interacting factors, including genetic, epigenetic, and environmental risk factors. Genetic studies may identify markers, which more accurately predict preterm birth than currently known risk factors, or novel proteins and/or pathways involved in the disorder. This review summarizes epidemiological and genetic studies to date, emphasizing the complexity of genetic influences on birth timing. While several candidate genes have been reportedly associated with the disorder, inconsistency across studies has been problematic. More systematic and unbiased genetic approaches are needed for future studies to examine the genetic etiology of human birth timing thoroughly. PMID- 18382884 TI - Pathophysiology, diagnosis and prognostic implications of endothelial dysfunction. AB - Endothelial dysfunction (ED) in the setting of cardiovascular risk factors such as hypercholesterolemia, hypertension, diabetes mellitus, chronic smoking as well as in patients with heart failure has been shown to be at least in part dependent on the production of reactive oxygen species (ROS) such as superoxide and the subsequent decrease in vascular bioavailability of nitric oxide (NO). Methods to quantify endothelial dysfunction include forearm plethysmography, flow-dependent dilation of the brachial artery, finger-pulse plethysmography, pulse curve analysis, and quantitative coronary angiography after intracoronary administration of the endothelium-dependent vasodilator acetylcholine. Superoxide sources include the NADPH oxidase, xanthine oxidase, and mitochondria. Superoxide produced by the NADPH oxidase may react with NO released by the endothelial nitric oxide synthase (eNOS) thereby generating peroxynitrite (ONOO-), leading to eNOS uncoupling and therefore eNOS-mediated superoxide production. The present review will discuss current concepts of how to assess endothelial function, prognostic implications of ED, mechanisms underlying ED with focus on oxidative stress and circulating biomarkers, which have been proposed to indicate endothelial dysfunction and/or damage, respectively. PMID- 18382885 TI - MicroRNAs: a new class of gene regulators. AB - Elucidation of the molecular basis of disease depends upon continued progress in defining the mechanisms by which genomic information is encoded and expressed. Transcription factor-mediated regulation of mRNA is clearly a major source of regulatory control and has been well studied. The more recent discovery of small RNAs as key regulators of gene function has introduced a new level and mechanism of regulation. Mammalian genomes contain hundreds of microRNAs (miRNAs) that each can potentially downregulate many target genes. This suggests a new source for broad control over gene regulation and has inspired extensive interest in defining miRNAs and their functions. Here, the identification of miRNAs, their biogenesis, and some examples of miRNA effects on biology and disease are reviewed and discussed. Emphasis is placed on the possible role for miRNA in nervous system development, function, and disease. PMID- 18382886 TI - High vitreous concentration of vascular endothelial growth factor in diabetic patients with proliferative retinopathy using statins. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) plays an important role in the development of diabetic retinopathy. Previous studies have suggested that angiotensin-converting enzyme (ACE) inhibitor therapy may reduce vitreous VEGF concentration in diabetic retinopathy. Also HMG CoA reductase inhibitors (statins), known for their beneficial effects on vascular endothelium, might influence vitreous VEGF concentration in diabetic retinopathy. AIM: Vitreous VEGF concentration of diabetic patients with proliferative retinopathy using statin therapy and/or ACE inhibitor therapy was studied. METHODS: Fifty diabetic patients with proliferative diabetic retinopathy, 21 diabetic control patients without proliferative retinopathy, and 43 non-diabetic control subjects underwent vitrectomy. Vitreous samples were collected at the beginning of surgery, and VEGF concentration was assessed using a chemiluminescent VEGF immunoassay. RESULTS: Vitreous VEGF concentration was significantly higher in diabetic patients with proliferative retinopathy using statins than in those not using statins. The diabetic patients with proliferative retinopathy were divided into subgroups according to use of ACE inhibitor and/or statin medication. These groups did not differ significantly in concentration of vitreous VEGF. CONCLUSIONS: Statin therapy is associated with high vitreous VEGF concentration in diabetic patients with proliferative retinopathy. In contrast to previous reports, ACE inhibitor use did not significantly influence vitreous VEGF concentration in these patients. PMID- 18382887 TI - Angiopoietin-2 levels as a biomarker of cardiovascular risk in patients with hypertension. AB - BACKGROUND: Abnormal angiogenesis is a pathophysiological component of cardiovascular disease (CVD), where circulating biomarkers of angiogenesis are associated with increased CVD risk in hypertension. We hypothesized that raised levels of angiopoietin (Ang)-1 and -2 would predict events in patients with hypertension treated for CVD. METHODS: We measured angiopoietin levels by enzyme linked immunosorbent assay (ELISA) in 251 hypertensive participants (85% male; mean age 63.5 years; 192 free of previous CVD events). Plasma angiopoietin levels were related to the subsequent CVD events over a mean follow-up period of 57.1 (SD 11) months. RESULTS: There were 11 cases of myocardial infarction (MI) and 18 cases of stroke during follow-up. Ang-2 was a significant predictor of MI, stroke, and composite CVD events, with the greatest event-free survival amongst those in the lower tertile (all P<0.05). Ang-1 was not predictive of CVD outcomes. Of CVD risk factors at recruitment (blood pressure, body mass index, plasma glucose, serum and high-density lipoprotein (HDL)-cholesterol), Ang-2 was the only discriminator of incident MI (area under curve (AUC) = 73%, P = 0.013), where a value >4.3 ng/mL optimized specificity and sensitivity. On Cox regression analysis (CVD treatments and risk factors), raised Ang-2 was an independent predictor of MI, P<0.05, but not stroke or composite outcomes. CONCLUSIONS: Among patients with hypertension, raised levels of Ang-2 were predictive of MI, and further study is warranted to evaluate the use of this biomarker in CVD management, risk stratification, and prevention. PMID- 18382888 TI - Lower economic status and inferior hygienic environment may protect against celiac disease. AB - BACKGROUND: The populations in adjacent Russian Karelia and Finland are equally exposed to grain products and share partly the same ancestry, but live in completely different socioeconomic environments. AIM: This creates an ideal epidemiological setting to study gene-environmental interactions in pathogenesis of celiac disease. METHODS: The prevalence of celiac disease and predisposing human leukocyte antigen (HLA) alleles was compared between Russian Karelia and Finland. Tissue transglutaminase antibodies and HLA-DQ alleles were screened from 1988 schoolchildren from Karelia and 3654 children from Finland. Children with transglutaminase antibodies were invited to small-bowel biopsy. Results. Transglutaminase antibodies were less frequent in Russian Karelia than in Finland (0.6% versus 1.4%, P = 0.005). Immunoglobulin class G (IgG) antigliadin antibodies were also less frequent in Russian Karelia (10.2% versus 28.3%, P<0.0001). Celiac disease was confirmed by duodenal biopsy in four of the eight transglutaminase antibody-positive Karelian children, giving a prevalence of 1 in 496 compared to 1 in 107 children in Finland. The same HLA-DQ alleles were associated with celiac disease and transglutaminase antibody positivity in both populations. CONCLUSIONS: The prevalence of transglutaminase antibodies and celiac disease is lower in Russian Karelia than in Finland. This may be associated with a protective environment characterized by inferior prosperity and standard of hygiene in Karelia. PMID- 18382889 TI - Senile systemic amyloidosis affects 25% of the very aged and associates with genetic variation in alpha2-macroglobulin and tau: a population-based autopsy study. AB - BACKGROUND: Senile systemic amyloidosis (SSA) is characterized by deposition of wild-type transthyretin (TTR)-based amyloid in parenchymal organs in elderly individuals. Previously, no population-based studies have been performed on SSA. METHODS: Here we have studied the prevalence and risk factors for SSA in a Finnish autopsied population aged 85 or over, as part of the population-based Vantaa 85+ Autopsy Study (n = 256). The diagnosis of SSA was based on histological examination of myocardial samples stained with Congo red and anti TTR immunohistochemistry. The genotype frequencies of 20 polymorphisms in 9 genes in subjects with and without SSA were compared. RESULTS: The prevalence of SSA was 25%. SSA was associated with age, myocardial infarctions, the G/G (Val/Val) genotype of the exon 24 polymorphism in the alpha2-macroglobulin (alpha2M), and the H2 haplotype of the tau gene (P-values 0.002, 0.004, 0.042, and 0.016). CONCLUSION: This population-based study shows that SSA is very common in old individuals, affecting one-quarter of people aged over 85 years. Myocardial infarctions and variation in the genes for alpha2M and tau may be associated with SSA. PMID- 18382890 TI - Homocysteine toxicity in connective tissue: theories, old and new. AB - Hyperhomocysteinemia causes connective tissue pathology. Several theories on the mechanism of homocysteine toxicity in connective tissue are reviewed briefly. A possible new mechanism was revealed recently in the discovery of a reaction in which homocysteine thiolactone is converted to mercaptopropionaldehyde. The reaction is the Strecker degradation of amino acids in which ninhydrin is replaced by the structurally similar dehydroascorbic acid. The reaction may occur in vivo and may be pathogenic to connective tissue in four ways: (1) the reaction may deplete ascorbic acid that is required for collagen synthesis, (2) the mercaptoaldehyde product may interfere with collagen synthesis, (3) the mercaptoaldehyde may cause abnormal cross-linking of collagen molecules, and (4) the mercaptoaldehyde may attach to collagen molecules rendering them antigenic and triggering an autoimmune response. PMID- 18382891 TI - MT3-MMP (MMP-16) is downregulated by in vitro cytokine stimulation of cartilage, but unaltered in naturally occurring equine osteoarthritis and osteochondrosis. AB - Matrix degradation by metalloproteinases is considered a key feature in the loss of articular cartilage seen in many joint diseases. Membrane-type matrix metalloproteinase-3 (MT3-MMP) expression is elevated in human cartilage in end stage osteoarthritis. We investigated whether MT3-MMP is similarly regulated in cartilage in two naturally occurring arthropathies in vivo and whether proinflammatory cytokines regulate its expression in vitro. MT3-MMP expression was evaluated in cartilage from horses with osteoarthritis and osteochondrosis and compared with age- and site-matched normal cartilage. MT3-MMP also was measured in normal cartilage stimulated with proinflammatory cytokines. MT3-MMP expression was not significantly altered in either osteoarthritis or osteochondrosis cartilage. However, gene expression was significantly downregulated by the addition of recombinant human interleukin-1beta, oncostatin M, or tumor necrosis factor-alpha to normal cartilage explants. The results suggest that MT3-MMP may not have a role in matrix destruction in equine cartilage diseases. Further work is required to characterize its regulation and function. PMID- 18382892 TI - Collagen XXIV (Col24a1) gene expression is a specific marker of osteoblast differentiation and bone formation. AB - Collagen XXIV is an ill-characterized fibrillar collagen that is predominantly expressed in the forming skeleton of the mouse embryo. Here we report that the Col24al gene is constitutively transcribed in the trabecular bone and periosteum of the newborn mouse as well. The bone specificity of Col24al was further documented using three well-characterized cell culture models of osteoblast differentiation. These in vitro analyses indicated that Col24al transcription is activated at about the same time as that of the osteocalcin gene, and gradually increases to eventually plateau as osteoblasts begin to deposit a mineralizing matrix. These findings lend further support to the hypothesis that collagen XXIV may be implicated in the formation of a mineralization-competent bone matrix. PMID- 18382893 TI - Effects of physical activities on biochemical and biomechanical properties of tendons in two commercial types of chickens. AB - The aim of our experiment was to study the effects of physical activities on the biochemical and biomechanical properties of tendons in 12 standard (S) broilers and 12 Label Rouge (LR) chickens. In the two types of birds no differences were found between control and active birds for body weights. Gastrocnemius (Gas) tendon and Pectoralis minor (Pm) tendons were harvested and processed for passive stretch tests prior to cooking or not. Some biochemical parameters also were determined. Results showed that total collagen content in Gas tendon was significantly higher in active than in control birds. However, no significant changes were found in collagen solubility in LR tendons while these values were increased in S ones. Active birds showed greater sGAGs content than control ones. Ultimate load was found to be significantly higher in active birds than in control. Deformability (defined by Poisson's ratio) of raw and heated at 80 degrees C Gas tendons increases in active groups because Poisson's ratio decreases. Physical activities also increase the rigidity (defined by elastic modulus) of raw and heated at 80 degrees C Gas tendons because elastic modulos values increase. Physical activity was not able to modify stiffness or maximum stress values in raw or heated at 80 degrees C Gas tendons from broilers whereas these two parameters were found to be slightly higher in active group from LR chickens only in raw tendons. All the biomechanical results recorded in Pm tendons from both types of chickens were not significantly different between control and active birds. A significant correlation was found between the total collagen content and stiffness in Gas tendon from LR active birds. PMID- 18382894 TI - Different crimp patterns in collagen fibrils relate to the subfibrillar arrangement. AB - Collagen fibril ultrastructure and course were examined in different connective tissues by PLM, SEM, TEM, and AFM. In tendons, collagen fibrils were large and heterogeneous with a straight subfibrillar arrangement. They ran densely packed, parallel, and straight changing their direction only in periodic crimps where fibrils showed a local deformation (fibrillar crimps). Other tissues such as aponeurosis, fascia communis, skin, aortic wall, and tendon and nerve sheaths showed thinner uniform fibrils with a helical subfibrillar arrangement. These fibrils appeared in parallel or helical arrangement following a wavy, undulating course. Ligaments showed large fibrils as in tendon, with fibrillar crimps but less packed. Thinner uniform-sized fibrils also were observed. Fibrillar crimps seem to be related to the subfibrillar arrangement being present only in large fibrils with a straight subfibrillar arrangement. These stiffer fibrils respond mainly to unidirectional tensional forces, whereas the flexible thinner fibrils with helical subfibrils can accommodate extreme curvatures without harm, thus responding to multidirectional loadings. PMID- 18382895 TI - A novel truncated TGF-beta receptor II downregulates collagen synthesis and TGF beta I secretion of keloid fibroblasts. AB - Hypertrophic scars and keloid are dermal proliferative disorders in wound healing. Transforming growth factor beta (TGF-beta) has been implicated in scar formation through the activation of fibroblasts and the acceleration of collagen deposition. Our study aimed to design a novel truncated (27-123 residues) type II TGF-beta receptor (tTGFbetaRII) and to determine its effects on the proliferation of keloid fibroblasts and the collagen synthesis as well as TGF-beta I expression of the cells. The coding sequences of TGF-beta I and tTGFbetaRII were amplified using RT-PCR and then cloned into pGBKT7 and pGADT7 vectors. A yeast two-hybrid experiment and a glutathione S-transferase (GST)-pull down assay were performed to verify the affinity of tTGFbetaRII to TGF-beta I. Our results indicated that treatment with tTGFbetaRII inhibited the growth of keloid fibroblasts and suppressed the synthesis of type I collagen in keloid fibroblasts in a concentration-dependent manner. Moreover, northern and western blot analysis revealed a decline of the TGF-beta I expression at both mRNA and protein levels after exposure to 5, 10 or 20 mug/ml of tTGFbetaRII. Together, our data suggested that the exogenous tTGFbetaRII can efficiently trap TGF-beta I from access to wild-type receptors and can suppress TGF-beta I triggered signals. Thus it may potentially be clinically applied to scar therapy. PMID- 18382896 TI - Bone mineral density and singh index predict bone mechanical properties of human femur. AB - The aim of our study was to assess the predictive value of the Singh index (SI), which estimates bone architecture, and bone density (BMD) when dealing with the mechanical competence of bone and to analze possible differences in bone properties between gender in humans. The relationship between SI, BMD, and mechanical competence was analyzed in 106 bone cylinders from 37 human femoral heads obtained during hip-joint replacement surgery for low energy fracture or for osteoarthritis. Bones from osteoporotic patients are less dense and more brittle compared with bones from osteoarthritic patients, as expected. Among osteoporotic patients female bones were more brittle than those from males, although BMD was similar. In osteoarthritic patients there were no significant differences among sexes. Bone mechanical competence varies according to BMD and to SI categories. Thus, our study suggests that bone strength is predicted by both BMD and bone architecture. PMID- 18382897 TI - Hyaluronan (HA) and serum-derived hyaluronan-associated protein (SHAP)-HA complex as predictive markers of cervical ripening in premature labor. AB - The purpose of this study is to investigate whether serum hyaluronan (HA) and serum-derived HA-associated proteins (SHAP)-HA complex predict cervical ripening and premature delivery. Sera were obtained from 64 women with normal pregnancies, 20 with full term delivery, and 13 with threatened premature labor. Concentrations of HA and SHAP-HA complex in serum were measured by sandwich ELISA. Serum concentrations of HA and SHAP-HA complex did not differ within first, second, and third trimester groups. The serum SHAP-HA complex was elevated in the full term labor group more than in the third trimester group; however, the concentrations of serum HA did not differ between both groups. The HA and SHAP-HA complex levels in sera were higher in the premature labor group than in the second trimester group. In the premature labor group, the SHAP-HA complex levels were higher in the cases with Bishop scores more than 4 points when compared with the cases with Bishop scores of 4 points or less. Increased levels of SHAP-HA complex in sera are possible predictive markers for cervical ripening in premature labor. PMID- 18382899 TI - Social epidemiology: strategies for public health activism. PMID- 18382898 TI - Falling between two stools; how a weak co-operation between the social security and the unemployment agencies obstructs rehabilitation of unemployed sick-listed persons. AB - PURPOSE: To explore significant factors behind the weak co-operation between local social insurance and unemployment agencies impairing the rehabilitation of unemployed sick-listed persons. METHOD: Individual, semi-structured face-to-face interviews with main actors directly involved in the sick-listing and rehabilitation process were conducted in the year 2000. In all 39 persons were interviewed: 25 professionals (physicians, public employment and social insurance officers) and 14 clients. Data were analysed according to Grounded theory method. RESULTS: The majority of unemployed sick-listed persons were declared too sick to work and were erased from the unemployment registers. This measure weakened the incentives for co-operation between the two main rehabilitation actors. The implication was that the unemployed sick-listed persons lost the opportunity of the co-ordinated rehabilitation they were entitled to. Three significant factors behind this process were identified by the main actors: indistinct regulation of co-operation, shifting political goals over time and conflicting goals between agencies; the last factor mainly a consequence of the other two. CONCLUSIONS: The findings suggest that labour market changes and manifest political goals influence the rehabilitation efforts giving low priority to difficult-to-place individuals such as unemployed sick-listed persons. In fact, a labour market problem turns into a medical problem. The hypothesis needs further testing in quantitative studies. PMID- 18382900 TI - Geropsychology - European perspectives for an aging world. PMID- 18382901 TI - Effect of Aerodiol administration on ocular arteries in postmenopausal women. AB - OBJECTIVE: The aim of the present study was to evaluate the alteration of ocular blood hemodynamics after intranasal administration of 17beta-estradiol, through measurement of the ophthalmic artery (OA) and the central retinal artery (CRA) using color duplex sonography. METHODS: Thirty healthy women who had been naturally postmenopausal for at least 1 year were enrolled in the study. A randomized, placebo-controlled, crossover, double-blinded study was conducted of the acute effect of 17beta-estradiol (Aerodiol; Servier, Chambrayles-Tours, France) on OA and CRA blood flow using color duplex sonography. RESULTS: The peak systolic and end diastolic velocities of the CRA were increased significantly after 17beta-estradiol administration compared with placebo. A significant decrease was also found in the pulsatility and resistive indices of the CRA. However, there were no statistically significant differences in OA flow velocities or pulsatility and resistive indices. CONCLUSIONS: Nasal 17beta estradiol administration in postmenopausal women causes an improvement in the ocular vascular Doppler indices. These results suggest that there could be a positive effect of Aerodiol on middle-sized arteries and arterioles. PMID- 18382902 TI - Women's perception of the efficacy of a soy extract with probiotic: the M3 study. AB - OBJECTIVES: To determine women's perception of soy extracts containing a probiotic on climacteric-related symptoms. DESIGN: A prospective multi-center survey. SUBJECTS AND METHODS: A total of 2044 women aged 40-65 years, who were recruited at random by a group of 198 Spanish gynecologists. The women completed a self-report survey at baseline and after 6 months' use of the soy extract (30 mg genistein and 30 mg daidzein) with probiotic (500 million spores of Lactobacillus sporogenes), calcium (141 mg) and vitamin D (5 microg). The questionnaire included 15 items on menopause and health and four about sexuality from the Cervantes scale. Patients and gynecologists completed an assessment about the clinical state. RESULTS: After 6 months, the studied soy extract improved climacteric symptoms and sexuality. Both patients and gynecologists were satisfied with the results. CONCLUSION: Peri- and postmenopausal patients usually experience common menopausal symptoms and many believe that soy extract improves their menopause symptoms. Physicians should be aware of patients' positive attitude to such products. PMID- 18382903 TI - Vascular endothelial growth factor level changes during human embryo development in culture medium. AB - OBJECTIVE: Implantation is a complex phenomenon consisting of the first strong contact between embryo and endometrium. Recent studies have demonstrated that this process is dependent not only on the 'readiness' of the endometrium, but also on complex interactions between endometrial and embryonic tissues that cross talk by means of different molecules (growth factors, cytokines, vasoactive factors). Investigations performed on human blastocysts indicate a role for vascular endothelial growth factor (VEGF) in these processes. The aim of the present study was to investigate VEGF levels at different stages in human embryo culture medium. STUDY DESIGN: We selected 20 women among patients undergoing assisted reproduction with the in vitro fertilization-blastocyst transfer protocol. The oocytes were inseminated by intracytoplasmic sperm injection. For each patient, approximately two cultures of four microinjected oocytes (and then of four embryos) were performed. Each culture of four oocytes/embryos was placed in one dish to increase the probability to detect small VEGF concentrations. RESULTS: Results showed significantly higher VEGF levels in the medium at blastocyst stage (12.16 +/- 2.80 pg/ml) compared with embryos at pronuclear stage (13.58 +/- 2.32 pg/ml) and microinjected oocytes (12.80 +/- 3.45 pg/ml). CONCLUSIONS: An important VEGF synthesis by blastocysts occurs during human embryo development. PMID- 18382904 TI - Prospective, randomized, comparative study of leuprorelin + human menopausal gonadotropins versus ganirelix + recombinant follicle-stimulating hormone in oocyte donors and pregnancy rates among the corresponding recipients. AB - AIM: To compare the clinical pregnancy rate in recipients of oocytes from donors treated with leuprorelin + human menopausal gonadotropins (hMG) with that obtained when the donors were treated with ganirelix + recombinant follicle stimulating hormone (rFSH). The secondary aim was to compare the donors' response to the two treatments. METHOD: A prospective, randomized, comparative study was conducted between January 2005 and November 2006 in a private hospital. Donors were randomized to receive a long protocol of leuprorelin + hMG (group DI) or ganirelix + rFSH (group DII). Their respective recipients were randomized to group RI or group RII, respectively. RESULTS: The characteristics of the donors were similar in both groups. More cycles were cancelled in group DI than in group DII (28.1% vs. 2.5%; p < 0.05). Compared with donors in group DII, the donors in group DI required a significantly higher dose of gonadotropins (2794 +/- 957 U vs. 1777 +/- 1043 U; p < 0.05) and more days of stimulation (11.7 +/- 2.3 vs. 9.5 +/- 1.5; p < 0.05); they also yielded fewer oocytes (15.0 +/- 6.1 vs. 17.9 +/- 8.6; p < 0.05). There were no differences in the characteristics of the recipients, in the fertilization rate or in the number of embryos transferred. The quality of transferred embryos was better in group RI (8.0 +/- 1.2 vs. 7.5 +/ 1.6; p < 0.05), and this group also achieved a better pregnancy rate per embryo transfer than did group RII (62.3% vs. 48.4%; p < 0.05). CONCLUSIONS: Treating oocyte donors with leuprorelin + hMG produces among recipients a greater probability of clinical pregnancy per embryo transfer than when donors are treated with ganirelix + rFSH; however, more cycles are cancelled and the former treatment is more unpleasant for donors. PMID- 18382905 TI - Increased rate of endometriosis and spontaneous abortion in an in vitro fertilization program: no correlation with epidemiological factors. AB - BACKGROUND: There are conflicting data concerning endometriosis and spontaneous abortion (SAB). The aim of the present study was to evaluate if there was any association between endometriosis and SAB. Moreover, we investigated risk factors in women with endometriosis and SAB. METHODS: The medical files of 457 married women with endometriosis and 200 infertile women without endometriosis were studied retrospectively. All cases were diagnosed by laparoscopy. Data concerning demographic variables and menstrual characteristics were recorded from 226 women with endometriosis, which were divided into two groups. Group 1 included 126 cases with endometriosis and SAB, and Group 2 comprised 100 parous women with endometriosis and without SAB. Statistical comparisons between groups were made using the chi(2) test and odds ratios (OR) and 95% confidence intervals (CI). RESULTS: The proportion of SAB was significantly higher in women with endometriosis than in infertile women without endometriosis (126/457 (27.6%) vs. 36/200 (18.0% ); OR = 1.7, 95% CI 1.1 = 2.6; p = 0.01). The frequency of nulligravid women was significantly higher in women with endometriosis than in the control group (OR = 1.9, 95% CI 1.4 - 2.81; p = 0.001). Mean age, age at onset of endometriosis, race, height, weight, body mass index, medical history of allergies, and family histories of endometriosis and cancer were similar in women with endometriosis and SAB and in parous women with endometriosis but without SAB. Moreover, the two groups were similar in age at menarche, length of cycle, duration and amount of flow, and the severity of disease. The incidence of infertility was significantly higher in women with SAB (p < 0.001). CONCLUSION: These data suggest but do not prove that the risk of SAB is increased in women with endometriosis. The epidemiological risk factors of endometriosis are not associated with an increase in the abortion rate. PMID- 18382906 TI - Differences in low-grade chronic inflammation and insulin resistance in women with previous gestational diabetes mellitus and women with polycystic ovary syndrome. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) and gestational diabetes mellitus (GDM) are both characterized by an increase in insulin resistance. Our goal in the present study was to measure insulin resistance (as estimated by homeostasis model assessment, sex hormone-binding globulin (SHBG) and adiponectin concentrations) and parameters of low-grade inflammation in non-diabetic, non hyperandrogenic ovulatory women with previous GDM (pGDM) and in non-diabetic women with classic PCOS, characterized by hyperandrogenism and oligo/anovulation. PATIENTS AND DESIGN: We evaluated 20 women with PCOS, 18 women with pGDM and 19 controls, all matched according to body mass index (BMI). Fasting blood samples were drawn in all women 3-6 days after spontaneous or dydrogesterone-induced withdrawal bleeding. Body fat distribution was assessed using dual-energy X-ray absorptiometry in all women. RESULTS: After adjusting for age and percent body fat, measures of insulin resistance such as SHBG and adiponectin concentrations were decreased and central obesity was increased in women with PCOS and pGDM compared with controls (all p < 0.05). Parameters of low-grade inflammation such as serum tumor necrosis factor-alpha and highly sensitive C-reactive protein concentrations, white blood cell and neutrophil count were increased only in women with PCOS compared with BMI-matched controls (all p < 0.05). CONCLUSIONS: Certain markers of insulin resistance are increased in both women with PCOS and women with pGDM, while low-grade inflammation is increased only in PCOS. PCOS and GDM might represent specific phenotypes of one disease entity with an increased risk of cardiovascular disease, whereby women with PCOS demonstrate an augmented cardiovascular risk profile. PMID- 18382907 TI - Effect of risedronate on biochemical marker of bone resorption in postmenopausal women with osteoporosis or osteopenia. AB - OBJECTIVE: The primary objective of the present study was to evaluate the effectiveness and adverse events of risedronate use in postmenopausal woman by measuring its effects on urinary crosslinked C-terminal telopeptides of type I collagen (CTx), a biochemical marker of bone resorption. METHODS: One hundred osteoporotic (control and treatment) and 111 osteopenic (control and treatment) postmenopausal women, selected according to World Health Organization criteria, were included in the study. The treatment groups (osteopenic and osteoporotic) were given risedronate 35 mg once a week. The primary endpoint was mean percentage change in CTx from baseline to 6 months. The secondary endpoints included evaluation of the incidence of clinical or laboratory adverse events occurring during the 6-month study period. The least significant change (LSC), calculated from the within-subject variability in the two control groups, was used to define response. RESULTS: Of the 211 women enrolled, 157 (74.4%) completed the study. After 6 months, urinary CTx levels were -54.7% (range -67% to -48%) below baseline in the osteoporotic treatment group and -66.7% (range 74% to -59%) below baseline in the osteopenic treatment group. Analysis of LSC showed that 89% of risedronate treatment groups were categorized as responders after 6 months of treatment. CONCLUSION: The study shows that osteoporotic and osteopenic women on risedronate treatment have statistically significant suppressed bone turnover and CTx can be useful to confirm this observation. The low withdrawal rate and adverse effects rate show that risedronate was well tolerated by the study population. PMID- 18382908 TI - Use of the Italian translation of the Female Sexual Function Index (FSFI) in routine gynecological practice. AB - AIMS: To investigate domains of sexual function in healthy women attending a gynecological office for routine annual check-up using the Italian translation of the Female Sexual Function Index (FSFI) according to age, reproductive status and hormonal treatments; and to confirm the usefulness of the FSFI in detecting relevant clinical entities. METHODS: Of 720 women (age range 18-65 years), 564 (78%) filled in a short anamnestic questionnaire and the FSFI assessing desire, arousal, lubrication, orgasm, satisfaction and pain. A semi-structured DSM-IV-TR clinical interview was administered to a convenience sample of women selected according to the quartile distribution of the median full scale FSFI score. Analysis of data was performed by frequency tables and non-parametric statistics. RESULTS: The median full scale score of FSFI in our study population was 27.6 (lower quartile: 18.7, upper quartile: 30.9) and the percentage of women under the lower quartile of the distribution was 24.4%. Sexual function decreased progressively with age, being significantly lower after 30 years and after 60 years (chi(2) = 52.6; p = 0.0001). Menopausal women had significantly lower median FSFI full scale score compared with fertile women and women who used oral contraception (OC) (p < 0.0001 for both), while users of hormone replacement therapy (HRT) displayed better overall sexual function than untreated postmenopausal women (p < 0.005). A positive diagnosis of female sexual dysfunction (FSD) was evident only in young women scoring under the lower quartile of the distribution (cut-off score: 23.4 for women not taking OC and 20.8 for OC users), while older women were dysfunctional also above the lower quartile of the distribution (cut-off score: 14.1 for menopause, 18.5 for HRT) of the FSFI full scale score. CONCLUSIONS: The FSFI is a powerful screening tool for FSD, especially in young fertile women, and may be used effectively in routine gynecological practice. PMID- 18382909 TI - Treatment of hirsutism with combined pill containing drospirenone. AB - The aim of the present study was to evaluate the biochemical and clinical effects of the combined pill consisting of ethinyl estradiol (EE) and drospirenone (DRSP) in hirsute patients. Fifty-two adolescents or young women from Greece were treated with 30 mug EE and 3 mg DRSP for 1 year. Hirsutism was evaluated by the Ferriman-Gallway (FG) score in the initial visit and at 3, 6 and 12 months. Follicle-stimulating hormone, luteinizing hormone, etradiol, free and total testosterone (T), sex hormone-binding globulin (SHBG), androstenedione and dehydroepiandrosterone sulfate were determined at the same intervals. Hirsutism improved and FG scores reduced to 77.4%, 67.2% and 51.8% at 3, 6 and 12 months, respectively. Plasma SHBG levels rose, while free and total T levels reduced from the third month onwards. In conclusion, the EE/DRSP pill improves hirsutism in women via antiandrogenic and antimineralocorticoid action. The biochemical manifestations of hyperandrogenism are also improved. PMID- 18382910 TI - Diagnosis of gestational diabetes mellitus by use of the homeostasis model assessment-insulin resistance index in the first trimester. AB - OBJECTIVE: Gestational diabetes mellitus (GDM) affects 2-10% of all pregnant women, causing increased morbidity and mortality, and is tested for in the second trimester of pregnancy. The purpose of the present study was to predict GDM in the first trimester. DESIGN AND METHODS: The study included 271 patients who were between the 10th and 14th week of gestation. Fasting glucose and insulin were measured in the first trimester and the homeostasis model assessment-insulin resistance index (HOMA-IR) was calculated for each patient. These values were compared with the results of the second-trimester glucose tolerance test. RESULTS: HOMA-IR values were higher in women with GDM. A cut-off value of 2.60 for HOMA-IR was calculated at the end of the study. CONCLUSION: Accepting patients whose HOMA-IR value is higher than 2.60 in the first trimester seems to be a good method to predict GDM. PMID- 18382911 TI - Sertoli-Leydig cell tumor: a rare ovarian neoplasm. Case report and review of literature. AB - BACKGROUND: Sertoli-Leydig cell tumor is a gonadal tumor of the sex cord-stromal type. It is a rare tumor comprising 0.1 to 0.5% of all ovarian tumors. Management of these cases poses a difficult therapeutic challenge. CASE: A 13-year-old girl presented with acute urinary retention, excessive hair growth and deepening of the voice. A mass the size of a 28-week gravid uterus was arising from the pelvis. Serum testosterone level was raised to 145.2 ng/dl. Computed tomography revealed a heterogeneously hypoechoic, solid cystic mass arising from the left adnexa. Left salpingo-oophorectomy was done. A histopathological diagnosis of Sertoli-Leydig cell tumor (intermediate, Meyers type II) was given. CONCLUSION: Patients with Sertoli-Leydig cell tumors present with signs of defeminization followed by masculinization. Age of the patient, stage of the disease and degree of tumor differentiation based on morphology are the most important factors to consider in the management of the case. PMID- 18382912 TI - Villages for depression care. PMID- 18382913 TI - Being delivered: spirituality in survivors of sexual violence. AB - A theoretical framework explaining how survivors of sexual violence use spirituality to respond to or recover from sexual violence is presented. Data were drawn from open-ended interviews of 27 women and 23 men who participated in a larger, ongoing study of women's and men's responses to sexual violence. Grounded theory methodology was used to develop the core category of Being Delivered, reflecting the participants' experiences of being rescued, saved, or set free from the effects of sexual violence by a spiritual being or power. The theoretical framework describing Being Delivered is composed of three dimensions: Spiritual Connection, Spiritual Journey, and Spiritual Transformation. The framework can be used by clinicians to guide discussions of spirituality and healing with survivors of sexual violence. PMID- 18382914 TI - Mental health service needs of a Latino population: a community-based participatory research project. AB - Community-based participatory research bridges the gap between academic researchers and the real-life issues of communities and offers promise for addressing racial and ethnic disparities in mental health care. The purpose of this community-based participatory research was to identify factors that affect access, use, and perception of mental health services by a Latino population at individual, organizational, and community levels. Individual level factors included health beliefs about mental illness and care, suspicions of providers, financial concerns, and culturally determined gender roles. Organizational factors included problems with access to care related to cost, lack of bilingual providers, and culturally competent care; and community level factors included distance between resources and the need for services to be provided in community sites. Immigration status and acculturation were identified as factors at all levels. PMID- 18382915 TI - Psychometric qualities of the coping response inventory-youth form. AB - This study examined the psychometric qualities of the Coping Response Inventory Youth (CRI-Y) form. Participants were 376 females and 248 males (age 15.9 + 1.0 year) recruited from four rural high schools in Western Pennsylvania who completed the CRI-Y as part of the instrument battery of a longitudinal study. Exploratory factor analysis was conducted to examine the factorial validity of the CRI-Y scale, and reliability coefficients were examined to address the reliability of the scale. The eight subscales of CRI-Y exhibited low to moderate reliability coefficients (from .47 to .70). The approach, avoidance, cognitive, and behavioral coping domains and total CRI-Y showed good reliability coefficients (.81, .87, .85, .84, and .91, respectively). Factor analysis of CRI Y subscales exhibited a unidimensionality of the subscales with a one-factor solution explaining 28%-40% of the total variance. PMID- 18382916 TI - Interdisciplinary collaboration: the need to revisit. AB - Given the complexity of the health care system and the prevalence of acute and chronic mental illnesses, multi-system interventions from an interdisciplinary team of health care providers are necessary to address the needs of individuals in psychiatric and mental health treatment venues. Despite claims that interdisciplinary practice is the norm, in reality it is difficult to accomplish (Martin-Rodriguez, Beaulieu, D'Amour, & Ferrada-Videla, 2005). Students must be taught the principles and elements of this way of providing care in order to succeed. This paper describes strategies for educating undergraduate nursing students to engage in interdisciplinary collaborative practice. Two case studies are used to illustrate how teaching collaborative skills can be integrated into a psychiatric undergraduate nursing course. PMID- 18382917 TI - Examining informed consent in persons with schizophrenia. AB - This descriptive study examined the responses of outpatients with schizophrenia to a tool designed to measure comprehension before obtaining informed consent to participate in a research study. We used the Evaluation to Sign Consent (ESC) form to document comprehension in 29 schizophrenia outpatients. Participants living in supervised housing were significantly more likely to require prompts than those living alone (chi(2) = 9.4, p = 0.024). Participants prescribed two antipsychotic medications were significantly more likely to require a prompt than those prescribed only one antipsychotic (chi(2) = 5.12, p = 0.023). Nurse researchers should consider documenting comprehension with the ESC due to its availability, time efficiency, and utility. PMID- 18382918 TI - Bullying, burnout, and mental health amongst Portuguese nurses. AB - Bullying is a concept commonly used to describe situations in which an individual is persistently treated in an abusive manner over a period of time, with a feeling of not being able to counterattack or defend him-or herself against the abuse. In this study we investigated both the prevalence of bullying in Portuguese nurses and the relationship between the symptoms of burnout and mental health in nurses who report being bullied. Participants were 107 nurses of the Portuguese Public Health System. We concluded that one out of six (13%) nurses had experienced bullying in the past six months. The three most common types of bullying behaviour experienced by the nurses were doing tasks below their level of competence, having areas of responsibility removed or replaced with more trivial or unpleasant tasks, and being exposed to unmanageable levels of workload. Bullied nurses had significantly higher levels of emotional exhaustion and lowered levels of mental health compared with non-bullied colleagues. PMID- 18382919 TI - Cultural context and treatment for substance-related disorders: an invitation. PMID- 18382921 TI - Process analytical technology: non-destructive assessment of anastrozole entrapment within PLGA microparticles by near infrared spectroscopy and chemical imaging. AB - The objective of this study was to evaluate near-infrared (NIR) spectroscopy and imaging as approaches to assess anastrozole entrapment within PLGA microparticles. By varying the polymer concentration, three batches containing the same amount of the drug were prepared. The spectral features that allow NIR drug quantitation were evaluated and compared with a best fit line algorithm. Actual entrapment efficiencies (EEF) determined via a destructive method were used for construction of calibration models using partial least square regression (PLS) or the algorithm. On the other hand, a chemical imaging system based on array detector technology was used to rapidly collect high contrast NIR images of the formulated microparticles. A quantitative measure of anastrozole entrapped was determined by calculating the percentage standard deviation of the distribution of pixel intensities in the PLS score images and histograms. Concerning conventional NIR analysis, both methods were equivalent for the prediction of EEF over the range of polymer levels studied. Correlation coefficients of more than 0.992 were obtained for either the calibration or prediction of EEF by the two methods; 0.392% and 0.374% were the standard errors of calibration and prediction (SEC and SEP) obtained for the prediction of EEF using the fit line, respectively, whereas the prediction of the EEF by the partial least square regression showed a SEC of 0.195% and SEP of 0.179%. As a result, the spectral best fit algorithm method compared favourably to the multivariate PLS method, but was easier to develop. In contrast, NIR spectral imaging was capable of clearly differentiating the three batches, both qualitatively and quantitatively. The percentage standard deviation increased progressively by increasing the ratio of drug-to-polymer concentrations. In conclusion, both NIR approaches were capable of accurate assessment of drug entrapment within microparticles. In addition, the NIR spectral imaging system provides a rapid approach for acquiring spatial and spectral information on microparticles. PMID- 18382922 TI - Polyurethane-based microparticles: formulation and influence of processes variables on its characteristics. AB - This study reports the development of polyurethane-based microparticles and the influence of some processes variables on its characteristics. These microparticles were prepared by emulsion polymerization, using poly(caprolactone) diol (PCL) and poly(propylene glycol), tolylene 2,4-diisocyanate terminated (TDI) or poly(propylene oxide)-based tri-isocyanated pre-polymer (TI). The reaction of polymerization was confirmed by attenuated total internal reflection Fourier transformed infrared spectroscopy (ATR-FTIR). Their thermal characteristics were investigated by dynamical mechanical thermal analysis (DMTA) and thermogravimetric analysis (TGA). For good microparticles formation, formulation 80/20 (mass ratio isocyanate/PCL) was the most indicated. Their spherical shape and smooth surface were observed by optical and scanning electron microscopy (SEM). Zeta potential measurements suggest that ionized carbonyl groups existent at the surface can be responsible for the negative potentials obtained. Respecting size and size distribution of the particles, measured by laser diffraction spectroscopy (LDS), the stirring speed and type were the process variables that most influenced it. PMID- 18382923 TI - Preparation and characterization of flurbiprofen-loaded poly(3-hydroxybutyrate-co 3-hydroxyvalerate) microspheres. AB - Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) microspheres containing flurbiprofen were prepared by an oil-in-water emulsion solvent evaporation method, in order to develop a particulate drug delivery system for localized administration. A response surface method (RSM) using a central composite design was employed to evaluate the effect of the poly(vinyl alcohol) (PVA) (%, w/v) concentration in the aqueous phase and the PHBV concentration in the organic phase (%, w/v) on some of the resulting microspheres properties. The response variables were the encapsulation efficiency (EE), the mean particle size, the width of particle size distribution (expressed by the SPAN value) and the required time for the in vitro release of 50% of the encapsulated drug (t(50%)). Second-order polynomial and linear equations were fitted to experimental data and were also used to interpret the results. Results indicated that the concentration of the stabilizer (PVA) showed a highly negative effect on the EE probably due to the increased drug solubility in the aqueous phase as a result of the higher PVA concentrations. Particle diameter mean size increased with the increased polymer concentration while the width of the particle size distribution was found to decrease with the increase of the stabilizer agent. Finally, results indicated that none of the investigated variables presented a significant effect on the t(50%) values. PMID- 18382924 TI - Enzymatic hydrolysis of organic-core microcapsules to produce aqueous-core microcapsules. AB - This paper describes the development of a new method to obtain aqueous-core microcapsules from organic-core capsules. The direct production of microcapsules, using tripropionin as organic material, followed by the hydrolysis of the core by a lipase was investigated. The enzymatic study showed that the enzyme obeyed a Michaelis-Menten mechanism and conditions for optimal activity were pH 7.5, 25-37 degrees C and 0% NaCl. Under these conditions, incubation of tripropionin alginate microcapsules in a buffer containing the enzyme successfully produced aqueous-core capsules with reduced accumulation of alginate in the core in approximately 3 h. PMID- 18382925 TI - Encapsulation and release of alpha-chymotrypsin from poly(glycerol adipate-co omega-pentadecalactone) microparticles. AB - Polymer-based microparticles are increasingly becoming of interest for a variety of applications including drug delivery. Recently poly(glycerol adipate) (PGA) and poly(glycol adipate-co-omega-pentadecalactone) have shown promise for delivery of dexamethasone phosphate and ibuprofen. In this paper the copolyester poly(glycol adipate-co-omega-pentadecalactone) was evaluated as a colloidal delivery system for encapsulated therapeutic proteins. Enzyme containing microparticles were prepared via the double water-in-oil-in-water (w/o/w) emulsion-solvent evaporation methodology. alpha-Chymotrypsin was used as a model proteolytic enzyme and its transfer was monitored during the emulsification process, in addition to in vitro release from formed particles. On average, 22.1 microg protein per 1 mg polymer was encapsulated, although gradual loss of activity of the protein, once released, was recorded. The work presented shows the potential of this polyester as a delivery system for enzymes via microparticles, with improvements to the system achievable via polymer and process optimization. The pendant hydroxyl groups on the polymer backbone provide future capacity for tailored alteration of the physical and chemical properties of the polymer, in addition to covalent attachment of various compounds. PMID- 18382926 TI - Polysulphone microcapsules containing silicone oil for the removal of toxic volatile organics from water. AB - Removal of volatile organic contaminants (VOCs) from water is an important task for environmental protection. Silicone oil has many merits but can not be directly applied in extraction of VOCs due to its high viscosity, which can negatively affect the mass transfer. Therefore a new purification method with extractant microcapsules has been developed in this work. Polysulphone microcapsules containing silicone oil were prepared successfully using a solvent evaporation method. The mass transfer performance was determined with toluene as the recovered component and with the prepared microcapsules as the separation agent. Extraction equilibrium could be reached in approximately 60 min. The maximum uptake of toluene was 24.5 mg g(-1) microcapsules. Microcapsules were reused four times and showed very good stability. Using microcapsules containing silicone oil as the separation agent, high and stable extraction capacity has been successfully achieved, especially without any phase dispersion and phase separation processes. It may be concluded that low energy will be required in the new separation process. PMID- 18382927 TI - Preparation and evaluation of stealth Tashinone IIA-loaded solid lipid nanoparticles: Influence of Poloxamer 188 coating on phagocytic uptake. AB - Stealth Tashinone IIA-loaded solid lipid nanoparticles (TA-SSLN) have been prepared by a nanoprecipitation/solvent diffusion method. Poloxamer 188 was used as a stealth agent. Nanoparticles were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The profile of Tashinone IIA release from TA-SSLN was studied. Phagocytosis was evaluated by incubating TA SSLN and non-stealth Tashinone IIA-loaded solid lipid nanoparticles (TA-NSLN) with murine macrophages. Human serum was added to evaluate opsonization of serum proteins. Rhodamine B was incorporated into nanoparticles as a fluorescent marker to observe and compare the phagocytic uptake of two kinds of nanopreticles. The results showed that TA-SSLN had an average diameter of (92.4 +/- 5.2) nm, zeta potential of (-20.4 +/- 1.3) mV, drug loading of (4.6 +/- 0.2)% and entrapment efficiency of (93.3 +/- 2.1)%. In vitro release experiment confirmed a sustained release of TA-SSLN and showed that the release of Tashinone IIA from TA-SSLN was in accordance with the Weibull equation. Phagocytosis studies showed significant differences between TA-SSLN and TA-NSLN and demonstrated that the Poloxamer 188 coating could decrease the macrophages uptake. PMID- 18382930 TI - Literature alerts. PMID- 18382931 TI - Plasma homocysteine levels in patients with beta-thalassaemia major. AB - OBJECTIVE: We investigated the level of homocysteine (HCY) and its relation with vitamin B12, folate and oxidative stress in patients with beta-thalassaemia major. MATERIAL AND METHODS: Plasma HCY, methionine, advanced oxidation protein products (AOPP) and serum vitamin B12, folate, ferritin and total antioxidant capacity (TAC) were determined in 32 thalassaemic patients and 27 control subjects. RESULTS: HCY (6.44+/-0.44 versus 8.71+/-0.57 micromol/L), methionine (12.57+/-1.8 versus 22.2+/-3.8 micromol/L), folate (9.14+/-0.48 versus 15.38+/ 0.71 nmol/L) and TAC (0.34+/-0.03 versus 0.56+/-0.03 mmol/L) significantly decreased in thalassaemic patients, whereas AOPP (20.26+/-1.8 versus 11.30+/-0.2 micromol/L) and ferritin (3481.0+/-512 versus 46.9+/-4.6 ng/mL) significantly increased. Vitamin B12 levels were similar in both groups (259.1+/-16.6 versus 228.9+/-7.4 pmol/L). CONCLUSIONS: These findings suggest that increased and uncompensated oxidative stress may lead to an increment in HCY catabolism in thalassaemic patients. PMID- 18382932 TI - Effects of long-term whole-body cold exposures on plasma concentrations of ACTH, beta-endorphin, cortisol, catecholamines and cytokines in healthy females. AB - OBJECTIVE: Cold therapy is used to relieve pain and inflammatory symptoms. The present study was designed to determine the influence of long-term regular exposure to acute cold temperature. Two types of exposure were studied: winter swimming in ice-cold water and whole-body cryotherapy. The outcome was investigated on humoral factors that may account for pain alleviation related to the exposures. MATERIAL AND METHODS: During the course of 12 weeks, 3 times a week, a group of healthy females (n = 10) was exposed to winter swimming (water 0 2 degrees C) for 20 s and another group (n = 10) to whole-body cryotherapy (air 110 degrees C) for 2 min in a special chamber. Blood specimens were drawn in weeks 1, 2, 4, 8 and 12, on a day when no cold exposure occurred (control specimens) and on a day of cold exposures (cold specimens) before the exposures (0 min), and thereafter at 5 and 35 min. RESULTS: Plasma ACTH and cortisol in weeks 4-12 on time-points 35 min were significantly lower than in week 1, probably due to habituation, suggesting that neither winter swimming nor whole body cryotherapy stimulated the pituitary-adrenal cortex axis. Plasma epinephrine was unchanged during both experiments, but norepinephrine showed significant 2 fold to 3-fold increases each time for 12 weeks after both cold exposures. Plasma IL-1-beta, IL-6 or TNF alpha did not show any changes after cold exposure. CONCLUSIONS: The main finding was the sustained cold-induced stimulation of norepinephrine, which was remarkably similar between exposures. The frequent increase in norepinephrine might have a role in pain alleviation in whole-body cryotherapy and winter swimming. PMID- 18382934 TI - Abstracts of the 10th Congress of the European Society of Contraception. April 30 May 3, 2008. Prague, Czech Republic. PMID- 18382933 TI - Evaluating influencing factors in estimation of renal function by extending the Bland-Altman approach. AB - Method comparison is frequently requested when newly developed methods are evaluated. In medical sciences in particular, numerous articles are published every year concerning new methods for improving diagnostics. Among several statistical approaches used to provide essential test characteristics, the Bland Altman method is well established, but actually provides no evaluation of possible factors influencing test quality. We propose an analysis of residuals obtained from the Bland-Altman data alleviating the determination of possible factors influencing accuracy and precision compared with different methods and give a short tutorial on application of Bland-Altman analysis. We illustrate this extension of the classical Bland-Altman method with an actual clinical example: estimation of renal function applying the common formula of Cockcroft and Gault. The influence of diabetes, gender and age on accuracy and precision of this estimation is analysed and inulin clearance is used as the gold standard. The subsequent analysis of residuals allows detection of the variables significantly influencing accuracy and precision. In the given example, diabetes and age, but not gender, are revealed as significant factors in glomerular filtration rate estimation when applying the Cockcroft formula. PMID- 18382939 TI - Imaging of the pelvis. PMID- 18382940 TI - Spectrum of MRI findings in clinical athletic pubalgia. AB - Athletic pubalgia is a frequently encountered syndrome for clinicians who treat active patients participating in a wide variety of athletic endeavors worldwide. Pathologies associated with this clinical scenario span anatomically from the pubic symphysis to the hip and include a myriad of poorly understood and incompletely described musculoskeletal entities, many of which are centered about the pubic symphysis and its tendinous attachments. In this article, we discuss the relevant anatomy and pathophysiology for the most frequently encountered of these disorders, using magnetic resonance (MR) images as a guide. We describe an MR imaging protocol tailored to clinical athletic pubalgia. We then review reproducible MRI patterns of pathology about the pubic symphysis, the rectus abdominis/adductor aponeurosis and the inguinal ring, as well as a group of clinically confounding entities remote from the symphysis but visible by MRI. PMID- 18382941 TI - Iliopsoas musculotendinous unit. AB - Hip pain is a common condition with many etiologies, and the workup often includes imaging. The iliopsoas musculotendinous unit is becoming more frequently recognized as the culprit for hip pain, partially due to the increase in athletic activity in the general population as well as to the increased demand in professional sports. Iliopsoas tendon injuries are also seen in the elderly population and documented as occurring following total hip replacement. Iliopsoas diseases, often overlooked as a cause of hip pain, include tendinosis, snapping tendon, bursitis, tears, and avulsion injuries. This article reviews the normal magnetic resonance imaging and sonographic anatomy of the iliopsoas musculotendinous unit and the imaging characteristics of diseases that involve the iliopsoas musculotendinous unit. We review the causes of a snapping hip and the role of sonography in evaluating and guiding treatment of the snapping iliopsoas tendon. PMID- 18382942 TI - Imaging of the hamstrings. AB - The hamstring muscles, located in the posterior thigh, include the biceps femoris, the semimembranosus, and the semitendinosus. The proximal portions of the hamstring muscles are subject to a variety of injuries and pathology. Many of these entities affect the origin of the hamstrings, including the tendinous enthesis, the underlying ischial tuberosity, and the surrounding tissues. Tendinosis and small partial tears at the origin are the result of chronic attrition. They may be accompanied by bursitis or hamstring syndrome. Apophysitis occurs in teenagers prior to complete fusion of the ischial apophysis and results from repeated traction injuries on the apophysis without discrete displacement. Abrupt injury at the origin from forced flexion of the hip results in osseous avulsions of the apophysis in teenagers and proximal tendon ruptures in adults. Other entities affect the muscles distal to the tendon origins. These injuries include strains and partial tears of the musculotendinous junction from acute indirect trauma, delayed onset muscle soreness from overuse of the muscle group without discrete remembered injury, and contusions and myositis ossificans from direct blunt impact. The imaging features of these injuries and pathology are fairly specific and diagnostic, with the exception of some cases of myositis ossificans and chronic ischial avulsions. PMID- 18382943 TI - Avulsion injuries of the pelvis. AB - Pelvic avulsion injuries occur in both the skeletally immature and adult patient populations. Avulsion injuries are most common in the adolescent age group and usually present as an avulsion of the unfused apophysis at the level of tendon attachment resulting from violent muscular contraction during an athletic endeavor. Acute apophyseal avulsion injuries are usually easily detected and adequately imaged with radiographs, but occasionally advanced imaging such as magnetic resonance imaging or computed tomography is required to detect and fully delineate the extent of injury. Chronic injuries can mimic a more aggressive lesion, but familiarity with the location of various tendon attachment sites on the osseous pelvis can help avoid this pitfall. Traumatic avulsion injuries can also occur in the skeletally mature pelvis; however, in the adult, the presence of a pelvic avulsion fracture in the absence of trauma should be considered a pathological lesion until proven otherwise. This article discusses the clinical and imaging findings of apophyseal avulsion injuries in the adolescent athlete, followed by a discussion of specific pelvis avulsion injuries that occur in the adult population. PMID- 18382944 TI - Anatomy, pathology, and MRI findings in the sports hernia. AB - "Sports hernia" is a frequently used term on athletic injury reports and in the sportscasting media, but its true definition remains elusive in the medical literature. Magnetic resonance imaging (MRI) is a useful tool in the evaluation of clinical athletic pubalgia, yet specific pathologies associated with this commonly encountered syndrome are poorly described in the imaging literature. In this article we review the musculoskeletal anatomy of the pubic region as well as several reproducible patterns of pathology on MRI we have encountered in patients with a clinical diagnosis of sports hernia. PMID- 18382945 TI - Imaging of stress injuries of the pelvis. AB - Stress fractures are common, representing the final stage in a continuum of bone response to continued mechanical damage. Encompassing fatigue- and insufficiency type fractures, stress fractures of the pelvis are likely underreported. Radiographs are insensitive to stress injuries, particularly those in the pelvis, whereas scintigraphy and magnetic resonance imaging are exquisitely sensitive. In this article we discuss the pathophysiology and imaging appearances of stress injuries of the pelvis and sacrum. Relevant literature regarding risk factors, problem-solving issues, and an imaging algorithm are discussed, with the goal of improving accuracy in the diagnosis of these common injuries. PMID- 18382946 TI - Sacroiliac joint imaging. AB - The sacroiliac (SI) joint has several unique anatomical features that make it one of the more challenging joints to image. The joint is difficult to profile well on radiographic views, and therefore the radiographic findings of sacroiliitis are often equivocal. Computed tomography images can usually show the findings of sacroiliitis and osteoarthritis earlier than radiographs. Magnetic resonance imaging performed with proper sequences is excellent for diagnosing even very early sacroiliitis and for following treatment response. The SI joint is often involved in patients with osteoarthritis or one of the inflammatory spondyloarthritides, most notably ankylosing spondylitis. Ankylosing spondylitis often presents with sacroiliitis, which appears as erosions, sclerosis, and joint space narrowing, eventually leading to ankylosis. Several disorders can cause sacroiliitis-like changes of the joint, including hyperparathyroidism and repetitive shear-stress injuries in athletes. The joint can become painful during pregnancy as it widens and develops increased motion, and some postpartum women develop iliac sclerosis adjacent to the joint termed osteitis condensans ilii. Another cause of SI joint pain is a disorder called sacroiliac joint dysfunction, which typically has few abnormal imaging findings. Patients with SI joint dysfunction, as well as sacroiliitis, often get relief from image-guided SI joint therapeutic injections. PMID- 18382947 TI - MRI of the pelvis: a guide to incidental findings for musculoskeletal radiologists. AB - Patients undergoing magnetic resonance imaging (MRI) of the pelvis for presumed musculoskeletal disease are commonly found to have abnormal imaging findings in the incidentally imaged pelvic viscera. Such incidental findings can be problematic for the musculoskeletal imager, both in terms of assigning clinical significance and determining the appropriate workup for a given abnormality. In this article we discuss the normal MRI appearance of the various pelvic organ systems as well as normal variants, emphasizing those that can mimic pathological processes. We then discuss the MRI appearance of common pathological entities encountered in these organ systems and the key imaging findings that should warrant a recommendation for further radiologic and/or clinical evaluation. PMID- 18382948 TI - [We need studies with relevant parameters!]. PMID- 18382949 TI - [Prevalence of smoking in women before and during pregnancy: population-based data]. AB - BACKGROUND AND OBJECTIVE: Population-based data on smoking behavior in Germany of women before or during pregnancy have been lacking. Smoking rates of these women have now been recorded over a period of 3 years against the background of growing tobacco control activities in Germany. METHODS: The study was conducted between 4/2003 and 3/2006 in the context of a prospective population-based survey about perinatal morbidity and mortality (Survey of Neonates in Pomerania - SNiP). This survey registers all newborns and their mothers in one defined region. Of the women eligible for the study 2 297 (68.1%) participated after delivery by giving data about their smoking behavior before and during pregnancy. RESULTS: 61.2% of the women had smoked at some time, 46.6% had smoked before, 24.2% into the 4. month and 20.5% into the last 4 weeks of pregnancy. Smoking rates remained unchanged over the 3 years that were studied. The rates of smokers who had quit by the time of delivery varied according to the length of school education: (<10 years: 30%; 10 y: 59%; >10 y: 84%) and age (<24 years: 45%; 25-30 y: 65%; >30 y: 77%). CONCLUSION: This study for the first time provides population-based data about the prevalence of smoking before and during pregnancy among women in Germany. The data show extremely high numbers of smokers, especially younger women and women of only 10 years or less of school education. Despite growing tobacco control activities no changes in smoking rates were observed over three years. These findings underline the need to develop effective interventions to prevent smoking of women before and during pregnancy. PMID- 18382950 TI - [Leiomyosarcoma of the inferior vena cava. Diagnosis using endoscopic ultrasound guided fine-needle aspiration biopsy]. AB - HISTORY AND ADMISSION FINDINGS: A 75-year-old woman was admitted because of upper abdominal pain, nausea and vomiting. INVESTIGATIONS: These symptoms were caused by a large tumor of the right retroperitoneal space. Computed tomography and transabdominal ultrasound did not unequivocally determine the site of the mass. Endoscopic ultrasound (EUS) and EUS-guided fine-needle aspiration biopsy (EUS FNA) ensured safe immunohistochemical diagnosis of a leiomyosarcoma of the inferior vena cava (IVC). TREATMENT AND COURSE: The patient underwent successful en-bloc resection of the tumor and the IVC followed by prosthetic reconstruction. CONCLUSIONS: This case emphasizes the role of EUS and EUS-FNA in the diagnosis of retroperitoneal lesions. Leiomyosarcoma of the IVC, although rare, should be considered in the differential diagnosis of a retroperitoneal mass. PMID- 18382951 TI - [Pelleted skull in multiple myeloma]. PMID- 18382952 TI - [Chaperono- and proteinopathies: molecular basis--novel therapeutic options]. PMID- 18382953 TI - [Complications after lung transplantation. Part 1: Intensive medical and pneumologic complications]. AB - Primary graft dysfunction and/or phrenic nerve lesion are common complications in the early phase after lung transplantation. Respiratory insufficiency is the most common reason for admission to the intensive care unit for patients after lung transplantation. Pulmonary complications in the post transplant management are acute allograft rejection (50% in the first year), infections, endobronchial bleeding, necrosis and fibrinous bronchitis. Chronic graft dysfunction (bronchiolitis obliterans syndrome) and infections are major obstacles to long term survival. PMID- 18382954 TI - [Ethical and legal aspects of including patients unable to consent in acute therapy studies. Example of a medication study for the treatment of intracerebral hemorrhage--the Heidelberg procedure]. AB - Clinical trials in patients who cannot sign an informed consent are only possible under certain circumstances. The present paper explains the legal prerequisites and ethic rationales, which may allow including patients in such a trial without having signed informed consent. Translation of these prerequisites into practice needs the implementation of special inclusion procedures. These procedures will be explained using the example of the recombinant factor VIIa (rFVIIa) trials for intracerebral hemorrhage. PMID- 18382955 TI - [Advertising with pictorial portrayal of people in professional clothing. Decision of the Federal Court from March, 1, 2007]. PMID- 18382956 TI - [How should endocarditis following heart valve replacement be treated?]. PMID- 18382957 TI - [Conceptual therapeutic approaches for joint disease]. PMID- 18382958 TI - [Controlled studies in German language publications]. PMID- 18382960 TI - [Surgical techniques. Interventions on the nose--interventions on the nasal septum II]. PMID- 18382962 TI - [Development of a nasal spray applicator]. PMID- 18382964 TI - [3D/ 4D-ultrasound in obstetrics--baby TV without diagnostics?]. PMID- 18382963 TI - [Sonography of the thyroid--part 2: thyroid inflammation, impairmant of thyroid function and interventions]. PMID- 18382965 TI - Hepatic transit time in benign liver lesions. AB - PURPOSE: Shortening of the hepatic transit time (HTT) of an echo enhancer indicates a potential metastatic spread to the liver. Previous studies showed a significant difference in the HTT between patients with metastases (6.5 sec) and patients without liver lesions (15 sec). The influence of benign liver lesions on the HTT is unclear, although benign liver tumors are a frequent finding. We therefore compared the HTT of benign liver lesions to the HTT of liver metastases. MATERIALS AND METHODS: Patients with focal liver lesions were enrolled in this study. We measured the HTT of an echo enhancer (Sonovue) in pulse inversion mode on the basis of time intensity curves (TIC). For this the hepatic artery and a branch of the hepatic vein were visualized simultaneously in an intercostal section. The difference between the arrival time of the echo enhancer in the hepatic artery and the hepatic vein offered the real HTT. RESULTS: 42 patients were enrolled in this study. 19 patients had metastatic growth in the liver, benign lesions were found in 23 patients. The mean HTT in patients with liver metastasis (6.4 sec +/- 1.6) was significantly less than in patients with benign lesions (10.4 sec +/- 3.4) (p < 0.001). None of the patients with liver metastasis had an HTT of greater than 10 sec. CONCLUSION: An HTT of greater than 10 sec seems to rule out liver metastasis. If the HTT is less than 10 sec, further evaluation of the liver is needed. There are indications that benign liver lesions also lead to a shortening of the HTT, but this is less pronounced than in patients with liver metastasis. PMID- 18382966 TI - Sonographical integrated test for detection of chromosomal aberrations. AB - PURPOSE: Different sonographical and biochemical methods for the detection of an elevated risk of chromosomal abnormality are used. The aim of our study was to establish a diagnostic scheme with the highest sensitivity by means of an algorithm incorporating all parameters. MATERIALS AND METHODS: In a group of 1490 pregnant women, ultrasound examination including nuchal translucency and nasal bone measurement was performed in the first trimester. Then, in the early second trimester, a second ultrasound examination combined with measuring nuchal thickness and nasal bone was carried out. RESULTS: The combination of the NT and NB measurement showed the highest sensitivity (94.7%), specificity (99.21%), PPV (85.7%) and NPV (99.73%). CONCLUSION: The proposed integrated test is characterised by a high predictive value for the detection of chromosomal abnormalities, low cost of performance and absolute safety for the foetus and could be offered to all pregnant women. The combination of two ultrasound examinations and biochemistry greatly increased the value of the test. PMID- 18382967 TI - [Quality standards of the DEGUM for performance of fetal echocardiography]. PMID- 18382968 TI - [Echocardiography: quo vadis?]. PMID- 18382969 TI - [Musician's medicine: when loudness becomes a problem]. PMID- 18382970 TI - Magnetic resonance imaging finding in severe head injury patients with normal computerized tomography. AB - AIM: Computerized tomography (CT) has been the primary diagnostic instrument of neurosurgical centers for head injury until now. However, there is a subgroup of patients who have persisting neurological deficits without visible pathology on CT. MATERIAL AND METHODS: Between 2000 and 2002, 3000 patients were enrolled in this prospective study in emergency unit. There was a total of 124 patients (4.1%) who had persisting severe neurological deficits with normal findings on repeated CT scans. These patients underwent 1.5 Tesla cranial MRI study between posttraumatic days 2 and 10 (5.2+/-4.5). Data on epidemiological, clinical, radiological factors and final outcome were collected. RESULTS: 76 (61.2%) of the patients were male and 48 (38.7%) were female. 113 (91.1%) of the 124 patients showed significant primary pathology on MRI study. The most common pathology was shear injuries in 75 (60.4%) followed by cortical contusion in 45 (36.2%) cases. 27 (21.7%) patients showed primary brain stem lesions and 8 (0.6%) patients showed subcortical gray matter lesions. 32 patients showed mixed lesions. There was one traumatic aneurysm detected. CONCLUSION: The MRI examination in head injury is informative in detecting subtle lesions when the neurological condition cannot be explained by CT. PMID- 18382971 TI - Role of diffusion-weighted MRG in predicting outcome in subarachnoid hemorrhage due to anterior communicating artery aneurysms. AB - AIM: Efficacy of DWI in detecting ischemic injury following anterior communicating artery aneurysmal SAH is studied MATERIAL AND METHODS: In this prospective study ,16 patients within 5 days of their ictus were included. Preoperative CT scan excluded an intracerebral infarct; an angiogram determined the extent of vasospasm; MR studies were done to detect cerebral ischemia and the diffusion coefficient (ADC) was calculated. Patients underwent surgery (n=12) or endovascular coiling (n=4). Postintervention CT scan within 24 hours and on day 5 determined radiological outcome as good or poor depending on the absence or presence of infarction. Clinical outcome at follow up (2-6 months) was regarded as good or poor when the patient was independent or dependent regarding daily activities, respectively,. RESULTS: Restricted diffusion occurred in 8 (80%) of poor and in 1 (16.6%) of good grade patients (p < 0.05). In patients with good radiological outcome (n=8), DWI restriction occurred in 2 (25%) patients. In patients with poor radiological outcome(n=8), DWI restriction occurred in 7 (87.5%) patients . At 2-6 month follow-up, 7 (77.8%) of 9 patients with restricted diffusion and 2 of 7 patients with no DWI abnormality showed a poor clinical outcome CONCLUSION: Clinical and radiological outcome at follow-up is related to the preoperative DWI. PMID- 18382972 TI - The accuracy and diagnostic yield of computerized tomography guided stereotactic biopsy in brain lesions. AB - OBJECTIVES: Radiological imaging techniques provide early detection of neurological diseases but they do not always provide an adequate and reliable diagnosis. With the help of stereotactic biopsy techniques, it is possible to access brain lesions safely and with high precision. We described the surgical method used in our clinic and discussed our results with the help of the current literature. METHODS: Ninety-four patients underwent computerized tomography guided stereotactic brain biopsy in our clinic. Anatomical locations, diagnostic yield and accuracy of the procedure, morbidity and mortality rates were analyzed. RESULTS: A total of 100 stereotactic surgery procedures were performed on 94 patients. The localizations of the lesions were 13.83% frontal, 21.27% temporal, 27.66% parietal, 4.25% occipital, 4.25% multiple, 27.66% deep seated and 1.06% suprasellar. The histopathological diagnoses were 61.71% neuro-epithelial tumors, 8.51% metastases and 10.64% infectious lesions. Diagnostic yield was 86.16% and the accuracy was 90% in our series. CONCLUSION: Computerized tomography-guided stereotactic brain biopsy is a reliable and safe method. Main diagnostic problems in SBB are tissue heterogeneity, insufficient material and sampling error. These problems can be minimized by careful correlation of clinical, radiological and histopathological findings by an experienced team and by using modern technologies. PMID- 18382973 TI - The segments and the inferior boundaries of the odontoid process of C2 based on the magnetic resonance imaging study. AB - AIM: The aims of this clinical study were to describe the segments and the inferior boundaries of the odontoid process as regards the embryological development of C2 based on the magnetic resonance imaging finding. MATERIAL AND METHODS: Cranial and cervical magnetic resonance images including occiput, C1, and C2, and those obtained for different reasons such as evaluation of cranial and upper cervical pathology were re-evaluated for this study. Synchondroses around the odontoid process were accepted as the boundaries between the neural arch and the body of C2. RESULTS: Thirty cases were included for this study. Fifteen of these were adult cases and the remaining 15 were in pediatric cases. Apicodental, dentocentral, neurocentral, and dentoneural synchondrotic articulations were clear, especially under the ages of 3 years. The dentocentral synchondrosis was found well below the line drawn through the level of superior articulating facets. CONCLUSION: This study demonstrated that the inferior boundary of the odontoid process is not located at the level of superior articulating facets. The real border between the odontoid process and the body of C2 is located well below the level of the superior articulating facets because of the location of the dentocentral synchondrotic articulation. This level should be considered in the classification of C2 fractures. PMID- 18382974 TI - Atypical teratoid/rhabdoid tumors: imaging findings of two cases and review of the literature. AB - Atypical teratoid/rhabdoid tumor (AT/RT) is a malignant embryonal central nervous system (CNS) tumor, manifesting in children, and composed of rhabdoid cells, with or without fields resembling a classical primitive neuroectodermal tumor (PNET), epithelial tissue and neoplastic mesenchyme. Around 200 cases of CNS AT/RT have been documented in the literature. Although the clinical and pathological findings have been defined in large series previously, and AT/RT has become increasingly recognized, awareness of typical AT/RT is important in making the correct diagnosis of this uncommon but probably underdiagnosed entity. Neuroradiologists rarely mention AT/RT in their differential diagnosis and this paper presents two additional cases in which clinical and pathological findings are combined with neuroradiological presentation. PMID- 18382975 TI - Basilar artery aneurysm associated with Behcet's Disease: a case report. AB - Vascular involvement is common in Behcet's Disease (BD). The venous system is often affected, while arterial system involvement is a rare occurrence. Intracranial aneurysm formation associated with BD is extremely rare. A 38- year old male patient with BD under the follow-up of the Dermatology department presented with acute headache. The cranial computed tomography revealed subarachnoid bleeding. A basilar artery aneurysm, which was ruptured during the procedure, was found on the cerebral angiography and the patient died the next day. This report adds our case to the literature of posterior cerebral circulation aneurysms associated with BD among the limited number of articles on this topic. PMID- 18382976 TI - Chronic subdural hematoma in capoeira sport. AB - Chronic subdural hematomas in young people is extremely rare and has some provoking factors such as V-P shunts, arachnoid cyst, anticoagulant drug usage, vigorous sports and coagulopathies. A static or dynamic mechanical load is almost always delivered to skull associated with either mild or severe head trauma. A 25 year old-man who was previously healthy has complained of intermittent headache for six months. He had been interested in capoiera (Brazilian exciting sport) for two years and has had no any evidence of head injury. After admission, he was operated immediately because of chronic subdural hematoma. We report a patient who is the first chronic subdural hematoma in the literature due to playing capoeira. PMID- 18382977 TI - A case report: a noninfantile desmoplastic astrocytoma. AB - INTRODUCTION AND CASE DESCRIPTION: Desmoplastic infantile astrocytomas (DIA) are rare intracranial tumors of infancy with distinctive clinical and radiological features. Despite their radiologically aggressive appearance they tend to follow a benign course with favorable prognosis even after subtotal resections. Non infantile cases are rarely encountered with only four cases reported before. The authors present a non-infantile DIA in a 4 years old female patient and discuss the clinical features, diagnosis, treatment of this rare entity. CONCLUSION: Although accepted as a tumor of infancy, DIA can also be encountered in older patients. Careful diagnosis and differentiation of DIA cases with other tumors, particularly malignant astrocytomas is important since the therapeutic strategies may differ. PMID- 18382979 TI - Should we operate all extradural spinal arachnoid cysts? Report of a case. AB - Spinal canal extradural arachnoid cyst is an uncommon, expanding lesion which may communicate with the subarachnoid space. They may be either intra or extradural with intradural cysts being less common. Spinal arachnoid cysts are usually in the thoracic spine, and they may cause symptoms due to spinal cord compression. Although surgery is the preferred way of management, there are reported cases with long term follow-up in the literature. Here we present a 14-year-old female who presented with intermittent low back pain for 1 year and a spinal arachnoid cyst that was followed for two years without any neurological deterioration. PMID- 18382978 TI - Endovascular treatment of primary infectious aneurysm in childhood: a case report. AB - Infectious aneurysms constitute 4% of all intracranial aneurysms. The microorganisms responsible are most commonly streptococcus viridans, staphylococcus aureus and combined bacterial infections. Nonetheless, cases with no reproduction in their cultures are rather frequent. A 6-year-old patient admitted with complaints of sudden headache, nausea, vomiting and high temperature. Intracerebral hematoma and saccular aneurysm located at the distal posterior cerebral artery were diagnosed as a result of the laboratory investigations and neuroradiological examinations. Infectious aneurysm was considered due to the clinical findings, morphology and location of the aneurysm. Although the causative microorganism was detected in blood culture, no focus could be detected. The aneurysm was hindered by endovascular intervention. In this manuscript, we discuss the infrequently seen childhood infectious aneurysm in the light of the pertinent literature. PMID- 18382980 TI - Venous outflow of the brain after bilateral complete jugular ligation. AB - A case of a patient with bilateral internal, external, posterior external and anterior jugular vein ligations and excisions performed in the neck due to a larynx tumor is presented. Radical neck dissection is a standard otorhinolaryngological procedure in the management of head and neck cancer patients with bilateral lymph node metastasis to the neck. Sacrifice of both internal and external jugular veins bilaterally has been recognized as a dangerous approach leading to intracranial hypertension with subsequent neurological sequela and death. In this report, we aimed to demonstrate how venous outflow from the brain diverts after jugular venous system obliteration. After bilateral jugular vein ligations, digital subtraction angiography (DSA) showed that the venous drainage route of the brain had been diverted from the jugular veins to the vertebral venous plexus. PMID- 18382981 TI - Arteriovenous malformation associated with multiple aneurysms including anterior communicating artery aneurysm located in the third ventricle: a case report. AB - Following the significant advances in neuroradiology, it has been now recognized that an intracranial arteriovenous malformation (AVM) lesion and vascular aneurysm(s) might coexist more often than estimated before. A58-yearold female seen due to a subarachnoid hemorrhage (SAH) was subsequently shown to have an AVM lesion fed by the pericallosal frontopolar, M1 segment of the middle cerebral artery along with three cerebral aneurysms. The AVM lesion was totally excised and the coexisting aneurysms were successfully clipped. As a very rare finding, one of the aneurysms originating from the Anterior Communicating Artery was localized in the 3rd ventricle penetrating the lamina terminalis. PMID- 18382982 TI - Esophageal carcinoma metastasis to cerebellum: a case report. AB - Esophageal carcinoma metastatic to the cerebellum and causing cerebellar compression is a rare entity. The authors report on the case of a 54-year-old woman in whom a metastatic cerebellar tumor secondary to esophageal carcinoma was treated. The patient presented with progressively increasing occipital headache. Computerized tomography scanning and magnetic resonance imaging results revealed a mass lesion in the cerebellum. Because of her previous esophageal carcinoma history, metastatic tumor to the cerebellum was assumed, and the patient underwent total resection of the tumor via median suboccipital approach. Histological examination of the specimen showed a metastatic adenocarcinoma of the esophagus. Postoperatively, the patient received 41 Gy radiation as adjuvant therapy. This case represents an addition to the reported literature of such a rare lesion. The diagnosis, treatment, and pathological findings of this case are presented and the pertinent literature is reviewed. PMID- 18382983 TI - Dural sinus thrombosis following head injury: report of two cases and review of the literature. AB - Dural sinus thrombosis (DST) usually involves the sagittal, transverse and sigmoid sinuses and is more common in women due to pregnancy, puerperium and oral contraceptive use. Other etiologies include coagulopathies, infection and head injury. We have present two DST cases following head injury. The first case was a 35-year-old man hospitalized because of one-week history of headache and repeated vomiting after a mild head injury. Thrombosis of the superior sagittal sinus, right transverse and sigmoid sinuses and right jugular vein was determined on angiography. The second case was a 25- year-old man operated on for epidural hematoma at the posterior fossa. Meningitis developed and an abducens palsy was determined. Magnetic resonance imaging demonstrated thrombosis of the right transverse and sigmoid sinus. Low molecular weight heparin was administrated for three months. Both cases had good recovery, but one had recanalisation of the thrombosis. Intracranial hematomas, depressed skull fracture or skull fracture that cross the sinus can obstruct the blood flow in the sinus. Moreover, closed head injury may cause to DST. Because of undefinitive pathophysiology, a consensus was not obtained on overall strategy concerning conservative, radiosurgical, or surgical therapy yet. PMID- 18382984 TI - Frontal bone and epidural tuberculosis. AB - Cranial bone and epidural tuberculosis is rare manifestation extrapulmonary tuberculosis. The incidence of tuberculosis of calvaria is on the rise in developing countries because of malnutrition, poor socioeconomic conditions, and immunodeficiency. We present the clinical features, radiology, histopathology, and surgical findings of a case of tuberculosis of the frontal bone with epidural extension. A 46 year-old female had presented with a history of painless right frontal swelling for the previous 4 months. The patient was operated on at a peripheral center for swelling in the right frontal region. In postoperative period, a leak that did not reply to nonspecific antibiotic therapy developed on the lesion. Right frontal epidural effusion was found in the patient who presented our clinic. Histopathological and microbiological examination suggested a diagnosis of tuberculosis. PMID- 18382985 TI - Hereditary neuropathy with liability to pressure palsies in a Turkish patient (HNPP): a rare cause of entrapment neuropathies in young adults. AB - Hereditary neuropathy with liability to pressure palsies (HNPP) is an autosomal dominant nerve disease usually caused by 1,5 Mb deletion on chromosome 17p11.2.2 p12, the region where the PMP-22 gene is located. The patients with HNPP usually have relapsing and remitting entrapment neuropathies due to compression. We present a 14-year-old male who had acute onset, right-sided ulnar nerve entrapment at the elbow. He had electrophysiological findings of bilateral ulnar nerve entrapments (more severe at the right side) at the elbow and bilateral median nerve entrapment at the wrist. Genetic tests of the patient demonstrated deletions in the 17p11.2 region. The patient underwent decompressive surgery for ulnar nerve entrapment at the elbow and completely recovered two months after the event. Although HNPP is extremely rare, it should be taken into consideration in young adults with entrapment neuropathies. PMID- 18382986 TI - Protein S deficiency, epileptic seizures, sagittal sinus thrombosis and hemorrhagic infarction after ingestion of dimenhydrinate. AB - Congenital protein S deficiency is associated with an increased risk of venous thrombosis. A14-year-old boy presented with epileptic seizures and thrombosis of the superior sagittal sinus and frontal hemorrhagic venous infarction after ingestion of 50 mg of dimenhydrinate, an antiemetic drug. The patient was found to be heterozygous for the factor V Leiden mutation and had a functional protein S deficiency. He recovered completely within a month after conservative treatment. Dimenhydrinate may have disrupted a subclinical pre-existing condition in this case. PMID- 18382987 TI - Cerebellar mutism following posterior fossa tumor resection in children. AB - AIM: Cerebellar mutism is a documented complication of posterior fossa surgery in pediatric ages. Risk factors such as the type of tumor, size, and location of tumor, hyrdrocephalus, postoperative cerebellar swelling for cerebellar mutism were investigated in this study. MATERIAL AND METHODS: A consecutive series of 32 children with a cerebellar tumor were operated on at the Haseki Educational and Research Hospital, Department of Neurosurgery, between 1990 and 2005. Their speech and neuroradiological studies were systematically analysed both preoperatively and postoperatively RESULTS: Cerebellar mutism developed in ten children (32%) in the early postoperative period. The type of tumor, midline localization, and vermian incision were significant single independent risk factors. In addition, an interdependency of possible risk factors (tumor > 5 cm, medulloblastoma) was found. The latency for the development of mutism ranged from 0 to 90 days (mean 15.6 d). The speech returned to normal in eight patients. All cases were accompanied by cerebellar ataxia. CONCLUSION: Mutism after posterior fossa tumor resection is also associated with ataxia. Cerebellar mutism usually has a self-limiting course and a favorable prognosis. PMID- 18382988 TI - Spontaneous extrusion of migrated ventriculoperitoneal shunt catheter through chest wall: a case report. AB - Ventriculoperitoneal (VP) shunt is the most commonly performed procedure for the management of hydrocephalus. VP shunt related complications remain a persistent problem in clinical practice. However, extrusion of components of shunt apparatus is very rare. Extrusion of ventriculo-peritonea l(VP) shunt catheter is an unusual complication of ventriculoperitoneal shunt Surgery. The authors report a case of a 17-year old female who presented with spontaneous extrusion of VP shunt catheter through the anterior chest wall. Pertinent literature is reviewed regarding the etiology and remedial measures to minimize this unusual complication of a very commonly performed neurosurgical procedure. PMID- 18382989 TI - A rare case of massive NF1 with invasion of entire spinal axis by neurofibromas: case report. AB - Neurofibromatosis type-1 (NF1) is a type of phakomatosis inherited in an autosomal dominant fashion. Also called 'von Recklinghausen disease' or 'peripheral neurofibromatosis', it comprises 90% of all neurofibromatosis (NF) cases. It is characterized by multiple peripheral nerve sheath tumors of benign character called neurofibromas. Surgical intervention is indicated when myelopathy and motor losses develop in the case of paraspinal neurofibromas, which are frequently localized to the cervical and lumbar regions. The level of surgical intervention required should be carefully considered and should take into account neurophysiological tests of paraspinal neurofibroma cases that allow estimation of the risk that the neurofibromas will invade the complete spinal axis. The best results are obtained with patients showing minimal neurological deficits during the pre-operative period. Little improvement may be expected from the patients who develop complete transection syndrome during the postoperative period. In the present paper, we discuss an NF1 case in which paraspinal neurofibromas were observed along the complete spinal axis of a 32-year-old male patient who arrived at the clinic reporting increasingly intense pins and needles and weakness. We discuss the patient's diagnosis, treatment, and prognosis, and relate this case to the literature. PMID- 18382990 TI - Bone development and its relation to fracture repair. The role of mesenchymal osteoblasts and surface osteoblasts. AB - Bone development occurs by two mechanisms: intramembranous bone formation and endochondral bone formation. Bone tissue forms by eventual differentiation of osteoprogenitor cells into either mesenchymal osteoblasts (MOBL), which synthesize woven bone in random orientation, or surface osteoblasts (SOBL), which synthesize bone on surfaces in a well oriented lamellar array. Bone repair uses the same formation patterns as bone development but the specific mechanism of repair is determined by the biomechanical environment provided. Bone synthesis and maintenance are highly dependent on the blood supply of bone and on cell-cell communication via the lacunar-canalicular system. Recent investigations highlight the molecular cascades leading to cell differentiation, the components of the structural proteins such as the various collagens, and tissue vascularization. The patterning of bone matrix from an initial woven to an eventual lamellar orientation is essential for bone to develop its maximum strength. This review demonstrates the repetitive nature of woven to lamellar bone formation as mediated by MOBLs and SOBLs in both normal vertebrate bones and bone repair. Repair, using endochondral, primary, direct and distraction osteogenesis mechanisms, is reviewed along with the associated molecular, vascular, and biophysical features. PMID- 18382991 TI - Prenatal diagnosis of clubfoot in low-risk population: associated anomalies and long-term outcome. AB - OBJECTIVES: To evaluate associated congenital anomaly risk, need for surgical treatment and long-term outcome in prenatally diagnosed clubfoot. METHODS: A retrospective study of 20 663 pregnant women who underwent routine ultrasound scanning at 18 to 22 weeks of gestation. Clubfoot was considered as complex or isolated if other structural or chromosomal abnormalities were also present or not. RESULTS: Forty-two cases of congenital clubfoot were diagnosed (incidence: 0.2%), 28 of them (66.6%) were isolated and 14 (33.3%) were complex, of which 3 (7.1%) had an abnormal karyotype and 11 (26.2%) had an associated structural anomaly. The false-positive rate was 2.3% (1 out of 32 liveborns). Out of the 41 confirmed affected fetuses, the defect was unilateral in 12 (29.3%) and bilateral in 29 (70.7%) cases. Surgery was necessary in 12 of the newborns (38.7%). The presence of a bilateral clubfoot was unrelated to either the presence of associated anomalies (p = 0.40) or to the necessity of surgery (p = 0.48). CONCLUSIONS: We provide outcome data about fetuses prenatally diagnosed for clubfoot. One-third are complex cases associated with other congenital anomalies. For isolated clubfoot, the risk of requiring surgery is about 40%. The detection of a bilateral defect does not worsen the prognosis. PMID- 18382992 TI - Monozygotic twins discordant for VACTERL association. PMID- 18382993 TI - Pathologic, radiographic and molecular findings in three fetuses diagnosed with HEM/Greenberg skeletal dysplasia. AB - BACKGROUND: Greenberg skeletal dysplasia is a very rare, autosomal recessive, in utero, lethal chondrodystrophy for which only eight index cases of diverse ethnic origin have been reported so far. The defect is associated with a defect in cholesterol biosynthesis and due to mutations in the gene encoding the lamin B receptor (LBR). METHODS: A familial case of three fetuses of a consanguineous Greek couple is presented including prenatal, physical, radiographic, histopathologic, and molecular genetic findings. RESULTS: The tentative diagnosis of Greenberg skeletal dysplasia based on pathological findings was confirmed by the identification of a homozygous, N547D amino acid substitution in the LBR gene in the third affected fetus. CONCLUSION: The present case represents the ninth described case of Greenberg dysplasia and the second case of Greek origin. The characteristic 'moth-eaten' radiographic appearance is already seen at 13 weeks' gestational age. PMID- 18382994 TI - Robotic-assisted gynaecological surgery-establishing training criteria; minimizing operative time and blood loss. AB - BACKGROUND: The objective was an evaluation of operative time and estimated blood loss (EBL) as a function of experience in gynaecological robotic surgery. METHOD: A retrospective analysis of 40 consecutive cases (approximately one case/week) over a 1 year period using the da Vinci) robotic system was performed, using data from two institutions, Newark Beth Israel Medical Center and Hackensack University Medical Center. Information was obtained from a single surgeon. Among the 40 cases there were 17 hysterectomies and 23 myomectomies. Each patient met the criteria of benign disease. In each institution, a da Vinci) system using three instrument arms and a camera arm was employed for every operation. RESULTS: Tests of differences in means were performed to compare the two groups. In group I (cases 1-20) the mean uterine volume was 863.0 cc and was similar to Group II (cases 21-40) at 632.6 cc. There was no significant difference between the groups when comparing blood loss; means were 86 cc for group I and 62.5 cc for group II. Operative time between groups, however, showed a significant difference (mean of 211.8 min for group 1 compared to 151 min for group 2; p < 0.05) and console time demonstrated a similar trend (mean for group 1 was 159.8 min compared to 90.8 min for group 2; p < 0.05). There were no conversions to laparotomy. Body mass index (BMI) and prior abdominal surgery were not significantly different. Multivariate regressions on operative time and EBL were performed, controlling for uterine weight and volume. The effect of experience on operative time was significant and negative; the coefficient on EBL was not significant. CONCLUSION: This study demonstrates statistical improvement in operative time after the first 20 cases for a single surgeon. This information could be used to establish criteria for training surgeons. PMID- 18382995 TI - Adapter for contact force sensing of the da Vinci robot. AB - BACKGROUND: At present, the da Vinci surgical robot system does not provide haptic feedback. One of the authors has proposed a contact-force sensing method called the 'overcoat method', in which the instrument/driver is supported by force sensors. In the da Vinci robot, the instrument jaws are powered by a wire pulley mechanism; thus, in order to apply the overcoat method to the da Vinci system, we must transfer the power through a frame that is supported by force sensors. METHODS: The authors have attempted to add a force-sensor function to the Sterile Adapter of the da Vinci system. In developing a sensorized adapter, a new configuration of force sensors and a new axial-force-free (AFF) joint have been devised in order to obtain an independent 'axial force effect' from the drive torque fed from the da Vinci robot arm. RESULTS: The force-sensing errors of the present system have been measured to have a maximum value of approximately 0.2 N while driving the jaws, and a maximum value of approximately 0.2 N when the robot arm is inclined with some excitation. Some impact reference forces applied on to the ends of the jaws agree with the outputs of the sensorized adapter to within <0.05 N. DISCUSSIONS: It is shown that the new adapter can be sterilized. One apprehension is that the total weight of the new adapter-approximately 1.2 kg might unbalance the robot arm. In the case of the new adapter, the centre-line of the instrument shaft is shifted externally through approximately 3.5 mm from its original position. However, a new cannula for the da Vinci robot might solve this problem. CONCLUSION: The new configuration of force sensors and the new AFF joint work well in their basic functions. The total force-sensing error is estimated as approximately 0.5 N. One of the main reasons for the error appears to be the deformation of the adapter frame. PMID- 18382996 TI - Structural heart defects associated with an increased nuchal translucency: 9 years experience in a referral centre. AB - OBJECTIVE: To investigate the congenital heart disease (CHD) found in association with an increased nuchal translucency (NT) at 11-14 weeks of gestation in chromosomally normal and abnormal fetuses. METHODS: Patients referred from January 1998 until May 2007 with an increased NT (> or = 95th percentile) where CHD was diagnosed were included. Chromosome analysis, fetal and postnatal echocardiography were performed. A postmortem examination followed pregnancy termination when possible. RESULTS: Major CHD was identified in 68 of 967 fetuses with an increased NT (median NT 4.8 mm, range 2.5-22 mm). Major CHD was found in 34 of 693 fetuses (4.9%) with a normal karyotype and increased NT (median 5.2 mm, range 2.5-9.6 mm). CHD frequency increased from 1.9%, with NT between 2.5 and 3.5 mm, to 27.7% when NT was > or = 6.5 mm. Septal defects predominated (20%) when NT was < or = 3.5 mm. With NT > 3.5 mm an equal distribution of CHD types was seen. Major CHD was identified in 34 of the 274 fetuses with an abnormal karyotype and increased NT (median 4.2 mm, range 2.5-22 mm). CONCLUSIONS: A variety of CHD is associated with an increased NT in the first trimester of pregnancy. Conotruncal defects, branchial arch derivative defects, left and right obstructive lesions (inflow and outflow) and shunts were seen. PMID- 18382997 TI - Prenatal sonographic imaging of Goldenhar syndrome associated with cystic eye. PMID- 18382998 TI - First-trimester ADAM12s as early markers of trisomy 21: a promise still unfulfilled? AB - BACKGROUND: Here we study the levels of ADAM12s prior to and after 10 weeks of gestation to establish further the effectiveness or otherwise of ADAM12s as an early screening marker. MATERIALS AND METHODS: Samples collected as part of routine first-trimester screening were retrieved from storage at - 80 degrees C. In total, 55 samples from pregnancies with trisomy 21 were identified, 31 collected between the 6th and 9th weeks of gestation and 24 collected after the 10th week. A series of 567 gestational age-matched samples collected during the same period formed the control group. RESULTS: The median, multiples of the median (MoM) ADAM12s, at a median gestation of 10.0 weeks was 0.66, which was significantly lower than in the controls (p = < 0.001) when compared by Mann Whitney test. The median MoM in those cases (n = 31) collected prior to 10 weeks was 0.618 MoM at a median gestation of 9.1 weeks. In those collected prior to 9 weeks (n = 14) the median was 0.596 at a median gestation of 8.6 weeks. CONCLUSIONS: The data from this study does not support data from a previous study showing ultra-low levels of ADAM12s in pregnancies affected by trisomy 21 prior to 10 weeks of gestation. PMID- 18382999 TI - Preliminary results of fetal Rhc examination in plasma of pregnant women with anti-c. AB - OBJECTIVES: Anti-Rhc antibodies may be the reason for the hemolytic disease of the newborn, therefore, noninvasive Rhc determination is important for pregnancy monitoring. For this purpose, we decided to introduce real-time polymerase chain reaction (PCR) method. METHODS: Blood from 200 donors, plasma and whole-blood from 11 Rhc-negative mothers, as well as blood from fathers and newborns were examined. Rhc sensitivity and specificity were first determined by real-time PCR using genomic DNA from donors. The same Rhc genotyping method was used for fetal Rhc detection in maternal plasma. To confirm the fetal Rhc-negative result, plasma was tested with a panel of biallelic insertion/deletion polymorphisms for the presence of fetal DNA. RESULTS: The c allele assay showed full specificity. The mean Ct value for one copy of c allele diluted in C-negative DNA was determined from extrapolating the correlation curve as 39.9. Full concordance was observed between the fetal Rhc genotypes from maternal plasma and the newborn phenotypes. CONCLUSIONS: Preliminary results show that it is possible to examine fetal c allele of RHCE gene in the plasma of pregnant women with anti-c by means of a noninvasive method. The diagnostic accuracy of the procedure, however, has yet to be confirmed in a larger group. PMID- 18383001 TI - NFkappaB2 gene duplication is associated with fetal pyelectasis in partial trisomy 10q (10q24.1 --> qter). PMID- 18383000 TI - The effects of acute treatment with paroxetine, amitriptyline, and placebo on driving performance and cognitive function in healthy Japanese subjects: a double blind crossover trial. AB - OBJECTIVE: To assess the effects of antidepressants on driving performance from a different methodological viewpoint in light of the recent traffic accidents. METHODS: In this double-blinded, 3-way crossover trial, 17 healthy males received acute doses of 10 mg paroxetine, 25 mg amitriptyline, and placebo. The subjects were administered three driving tasks--road tracking, car following, and harsh braking--performed using a driving simulator and three cognitive tasks--Wisconsin Card Sorting Test, Continuous Performance Test, and N-back test at baseline and at 1 h and 4 h post-dosing. The Stanford Sleepiness Scale scores were also assessed. RESULTS: At 4 h post-dosing, amitriptyline significantly impaired road tracking and car-following performance, reduced driver vigilance, and caused subjective somnolence. Paroxetine impaired neither driving performance nor cognitive function. CONCLUSIONS: Acute doses of amitriptyline significantly impaired driving performance in the context of driving on crowded urban roads at relatively low speeds. This setting is important with respect to skills necessary for daily driving and may be difficult to measure in actual driving tests. This simulator-based study replicated the results of previous studies and could be considered complementary to them. This method may enable easy and safe screening of the driving hazard potential of drugs. PMID- 18383002 TI - Bone regeneration induced by adenoviral vectors carrying til-1/Cbfa1 genes implanted with biodegradable porous materials in animal models of osteonecrosis of the femoral head. AB - A new approach for bone regeneration is needed for idiopathic osteonecrosis of the femoral head (ION). Core binding factor alpha1 (Cbfa1) was reported in 1997 as the most important transcription factor for osteoblastic differentiation. The transgenics of transcription factors affecting bone formation might be useful tools for the bone regeneration. The purpose of this study was to investigate the effects of the implantation of adenoviral vectors carrying Cbfa1 genes implanted with biodegradable porous materials on bone formation in an animal model of ION. Robust and rapid bone regeneration in large bone defects was achieved with the implantation of adenoviral vectors carrying Cbfa1 genes. These results suggest that the Cbfa1 genes induce a rapid osteoblastic differentiation and the biodegradable scaffold successfully functioned as a delivery vehicle for the Cbfa1 gene, as they induced osteogenic repair in vivo, even in necrotic bone. PMID- 18383003 TI - Delusional beliefs in psychotic depression vary according to age of onset. PMID- 18383004 TI - Allergic contact dermatitis to topical drugs--epidemiological risk assessment. AB - PURPOSE: Estimation of the population-based relative incidence (RI) of contact sensitization to a set of topical drug allergens (cases/100,000 defined daily doses (DDDs) per year) (1995-2004) and comparison of the RI for drugs with a similar therapeutic scope. METHODS: Clinical data regarding the frequency of contact sensitization to important topical drug allergens in Germany were obtained from the Information Network of Departments of Dermatology (IVDK). This was extrapolated to the general population level using the 'clinical epidemiology and drug utilization research' (CE-DUR) approach. As denominator of exposure, national prescription data (DDDs of topical drug specialties) provided by the WIdO Research Institute (Bonn) were aggregated per substance. RESULTS: The probable RIs of contact sensitization to topical 'skin' corticosteroids ranged from 0.3 (dexamethasone phosphate disodium salt) to 23.3 (amcinonide) cases/100,000 DDDs per year. Concerning topical aminoglycoside antibiotics, the RI of framycetin sulphate was about threefold higher than that of gentamicin sulphate. Regarding topical ophthalmic use, the RI of kanamycin sulphate was higher compared to gentamicin sulphate. Active principles marketed over-the counter (OTC) had, in general, lower RIs, with the exception of bufexamac, benzocaine, clioquinol and phenylephrine. CONCLUSIONS: The population-based risk assessment--quantitatively considering exposure in the RI estimation--revealed a ranking of contact sensitization risk to topical drugs which partly differed from the respective frequencies in the clinical patch test population. Some drugs available OTC carry non-negligible risk, too. The current findings should contribute to differential therapeutic considerations regarding topical drug use. PMID- 18383005 TI - Identification of oxidized proteins in rat plasma using avidin chromatography and tandem mass spectrometry. AB - The objective in much of the proteomics literature today is to establish the difference between healthy and disease states at the protein level using blood plasma. A critical component in this endeavor is to establish what is normal. The focus of the work reported here was to do this with oxidized proteins that might relate to oxidative stress and oxidative stress-related diseases. Oxidative stress is known to increase markedly in cancer, diabetes, heart disease, and neurodegenerative diseases. Since proteins are one of the targets of ROS, generated by oxidative stress, oxidized proteins are excellent biomarker candidates for these diseases. But first it is necessary to identify oxidized proteins that occur in the healthy state. Healthy rat plasma was used in this study as a source for the identification of naturally oxidized proteins. Freshly drawn blood was treated with biotin hydrazide to selectively derivatize carbonyl groups in oxidized proteins. Oxidized proteins thus biotinylated were separated from the other plasma proteins using avidin affinity chromatography. Affinity selected proteins were further fractionated on a C(8) RP column and fractions collected. The collected fractions were then tryptic digested and the peptides identified using a combination of LC/MS/MS and database searches. One hundred forty-six proteins were identified using 700 signature peptides from the tryptic digested chromatographic fractions. The most frequently encountered proteins in the samples were keratins. Brain and liver were among the organs contributing the most oxidized proteins to plasma followed by heart and kidney. PMID- 18383006 TI - Profiling the metabolic proteome of bovine mammary tissue. AB - 2-DE and MALDI mass fingerprinting were used to analyse mammary tissue from lactating Friesian cows. The goal was detection of enzymes in metabolic pathways for synthesis of milk molecules including fatty acids and lactose. Of 418 protein spots analysed by PMF, 328 were matched to database sequences, resulting in 215 unique proteins. We detected 11 out of the 15 enzymes in the direct pathways for conversion of glucose to fatty acids, two of the pentose phosphate pathway enzymes and two of the enzymes for lactose synthesis from glucose. We did not detect enzymes that catalyse the first three reactions of glycolysis. Our results are typical of enzyme detection using 2-DE of mammalian tissues. We therefore advocate caution when relating enzyme abundances measured by 2-DE to metabolic output as not all relevant proteins are detected. 2-D DIGE was used to measure interindividual variation in enzyme abundance from eight animals. We extracted relative protein abundances from 2-D DIGE data and used a logratio transformation that is appropriate for compositional data of the kind represented in many proteomics experiments. Coefficients of variation for abundances of detected enzymes were 3-8%. We recommend use of this transformation for DIGE and other compositional data. PMID- 18383008 TI - Treatment of missing values for multivariate statistical analysis of gel-based proteomics data. AB - The presence of missing values in gel-based proteomics data represents a real challenge if an objective statistical analysis is pursued. Different methods to handle missing values were evaluated and their influence is discussed on the selection of important proteins through multivariate techniques. The evaluated methods consisted of directly dealing with them during the multivariate analysis with the nonlinear estimation by iterative partial least squares (NIPALS) algorithm or imputing them by using either k-nearest neighbor or Bayesian principal component analysis (BPCA) before carrying out the multivariate analysis. These techniques were applied to data obtained from gels stained with classical postrunning dyes and from DIGE gels. Before applying the multivariate techniques, the normality and homoscedasticity assumptions on which parametric tests are based on were tested in order to perform a sound statistical analysis. From the three tested methods to handle missing values in our datasets, BPCA imputation of missing values showed to be the most consistent method. PMID- 18383009 TI - Characterizing the dynamic nature of the Yersinia pestis periplasmic proteome in response to nutrient exhaustion and temperature change. AB - The periplasmic proteome of Yersinia pestis strain KIM6+ was characterized using differential 2-DE display of proteins isolated from several subcellular fractions. Circa 160 proteins were designated as periplasmic, including 62 (putative) solute-binding proteins of ATP-binding cassette (ABC) transporters (SBPs) and 46 (putative) metabolic enzymes. More than 30 SBPs were significantly increased in abundance during stationary phase cell growth, compared to the exponential phase. The data suggest that nutrient exhaustion in the stationary phase triggers cellular responses resulting in the induced expression of numerous ABC transporters, which are responsible for the import of solutes/nutrients. Limited availability of inorganic phosphate (P(i)) also caused dramatic proteomic changes. Nine proteins were functionally linked to the mobilization and import of three small molecules (P(i), phosphonate and glycerol-3-phosphate) and accounted for nearly half of the total protein mass in the periplasm of P(i)-starved cells. When cells were grown at 26 degrees C versus 37 degrees C, corresponding to ambient temperatures in the flea vector and mammalian hosts, respectively, several periplasmic proteins with no known roles in the Y. pestis life cycle were strongly altered in abundance. This included a putative nitrate/sulfonate/bicarbonate-specific SBP (Y1004), encoded by the virulence associated plasmid pMT1 and increased in abundance at 37 degrees C. PMID- 18383010 TI - Proteomic analysis on a high salt tolerance introgression strain of Triticum aestivum/Thinopyrum ponticum. AB - Soil salinity is a major abiotic constraint to agricultural productivity. We successfully bred a new common wheat (Triticum aestivum L.) introgression variety (Shanrong No. 3) with high salt-tolerance via asymmetric somatic hybridization between common wheat cultivar (Jinan 177) and UV-irradiated Agropyron elongatum (Thinopyrum ponticum Podp). We report here a comparative proteomic analysis to investigate variety-specific and salt-responsive proteins between seedling-roots of Shanrong No. 3 and Jinan 177. In total, 114 spots reproducibly presented differential expression patterns on 2-DE maps. Of them, 34 were variety-specific and 49 were salt-responsive. We identified 110 spots by MALDI-TOF MS and partially confirmed by MALDI-TOF-TOF MS, and functionally classified them into signal transduction, transcription and translation, transporting, chaperones, proteolysis and detoxification, etc. Meanwhile, we also found the alteration of protein expression of Shanrong No. 3 through inhibition of old proteins and production of novel ones, change in abundance and sensitivity of some nonsalt responsive and salt-responsive proteins, as well as PTMs. Furthermore, comparison between proteome and transcripteome using cDNA microarray showed that there were only 20 proteins with abundances correlative to signal densities of corresponding EST probes. This study gives us a global insight into proteomic difference between Shanrong No. 3 and Jinan 177 in constitute and to salt-response. PMID- 18383011 TI - Affinity capillary electrophoresis for identification and investigation of human Gc-globulin (vitamin D-binding protein) and its isoforms interacting with G actin. AB - A CE procedure was established for the nondenaturing separation and identification of the isoforms of the actin-binding human plasma protein Gc globulin. To characterize interactions with globular actin (G-actin), a novel method was developed for the simultaneous qualitative assessment of the binding interaction between the three major isoforms of Gc-globulin and G-actin using pre equilibrium affinity CE and UV detection. Evidence was found that some difference in binding affinity existed among the isoforms, although the quantification of this difference was not feasible by UV detection because of the high affinity nature of the binding. The difference in affinity appeared to be related to the pI of the isoforms; a high pI corresponding to a high affinity. For quantitative binding studies Gc-globulin was fluorescently labeled with 5-(and-6) carboxyfluorescein, succinimidyl ester (CFSE). Data suggested that extensive labeling interfered with actin binding but with moderately labeled Gc-globulin it was possible to determine a dissociation constant of K(d) = 21 +/- 1 nM for the binding between labeled Gc-globulin and G-actin using pre-equilibrium affinity CE and LIF detection. PMID- 18383012 TI - Determination of ethyl glucuronide in human serum by hyphenation of capillary isotachophoresis and zone electrophoresis. AB - The determination of ethyl glucuronide (EtG), a marker of recent alcohol consumption, in human serum by hyphenation of capillary ITP (CITP) and CZE is reported. For CITP step, 1 x 10(-2) M hydrochloric acid adjusted with epsilon aminocaproic acid (EACA) to pH 4.4 was used as the leading electrolyte, and 1 x 10(-2) M nicotinic acid with EACA, pH 4.4, was used as the terminating electrolyte (TE). All electrolytes contained 0.2% hydroxypropylcellulose to suppress electroosmosis. In CITP, EtG was separated from fast serum macrocomponents chloride, phosphate, lactate, and acetate. Zones of microcomponents including EtG that migrated between acetate and nicotinate were forwarded to the second capillary filled with a BGE identical with the TE. Conductivity detection was used in the CITP step. Sensitive detection in the CZE step was performed using indirect spectrophotometric detection at 254 nm. The assay is based on a 1:5 dilution of serum with deionized water and has a concentration LOD for EtG in diluted sample of 9.8 x 10(-9) M. The method was used for the determination of EtG in sera of volunteers consuming alcohol. PMID- 18383013 TI - Novel quantitative electrophoretic analysis of endotoxins on microchips. AB - A novel microchip electrophoresis method was developed and applied for sensitive detection and quantitative analysis of endotoxins extracted from Gram-negative bacteria. The method provides a fast and quantitative differentiation of smooth and rough endotoxins based on the solubilization and complexation of the lipopolysaccharides with dodecylsulfate, and then with a fluorescent dye. The migration of the complexes was followed by LIF detection. The novel method is able to replace the SDS-PAGE with the advantage of high speed and better sensitivity, and by avoiding the laborious gel-preparation and silver staining. PMID- 18383014 TI - Enzymatic in-capillary derivatization for glucose determination by electrophoresis with spectrophotometric detection. AB - The following paper compares several procedures of in-capillary bienzymatic derivatization with regard to glucose determination with the use of glucose oxidase and horseradish peroxidase. The procedures discussed below include continuous contact in the capillary, plug-plug injection, and sequential injection with incubation in the capillary inlet. The reaction of hydrogen peroxide catalyzed by peroxidase was performed using two different substrates. The best results were achieved for nicotinamide adenine dinucleotide, reduced disodium salt (NADH) acting both as a chromogenic reagent and a substrate for peroxidase, while the method employed was sequential injection and incubation at the capillary inlet. The LOD was estimated to be 25 nM with a linear response up to 0.1 microM. PMID- 18383015 TI - Rapid capillary electrophoresis time-of-flight mass spectrometry separations of peptides and proteins using a monoquaternarized piperazine compound (M7C4I) for capillary coatings. AB - A monoquaternarized piperazine, 1-(4-iodobutyl) 4-aza-1-azoniabicyclo[2,2,2] octane iodide (M7C4I), has been evaluated as a surface derivatization reagent for CE in combination with TOF MS for the analysis of proteins, peptides, and protein digests. The M7C4I piperazine, at alkaline pH, forms a covalent bond via alkylation of the ionized silanols producing a cationic surface with a highly stable and reversed EOF. The obtained surface yields rapid separations (less than 5 min) of peptides and proteins at acidic pH with high separation efficiencies (up to 1.1 x 10(6) plates/m for peptides and up to 1.8 x 10(6) plates/m for proteins) and no observed bleeding of the coating reagent into the mass spectrometer. The simplicity of the coating procedure also enables fast (2 min) regeneration of the surface, if necessary. This is useful in the analysis of complex samples in order to prevent possible memory effects. The potential of using M7C4I-coated capillaries for MS analysis of complex samples is demonstrated by the separation of peptides, proteins, and protein digests. Even more, the spectacular thing in which large intact proteins with molecular masses over 0.5 MDa could be separated. The coating showed good ability to handle these large proteins with high efficiency and retained peak shape as demonstrated by separation of IgG(1) (150 kDa) and thyroglobulin (669 kDa). PMID- 18383017 TI - Use of quasi-isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances. AB - The use of quasi-isoelectric anolytes and catholytes has been investigated to improve CIEF performances. Narrow pH cuts of carrier ampholytes (NC) have been compared to more conventional couples of anolytes/catholytes (phosphoric acid/sodium hydroxide and glutamic acid/lysine). First, a CIEF setup that consists in a bare silica capillary and 70:30 water/glycerol separation medium has been used. The experiments have shown that when using NC instead of more classical anolytes and catholytes, an increase in the protein detection time was observed and the resolutions obtained for neutral and acidic proteins were doubled. Moreover, according to the NC fraction used, the resolution was modified. In order to investigate further the mechanisms involved, a second setup using a capillary coated with hydroxypropylcellulose was used. With this setup no difference has been observed when changing anolyte and catholyte nature. A simple methodology has then been developed to evaluate EOF during focusing and mobilization steps of CIEF experiments. It highlighted the crucial role played by EOF when using a bare silica capillary. EOF indeed decreased by 33% during mobilization step when using NC instead of classical anolytes and catholytes. PMID- 18383016 TI - Two-dimensional strong cation exchange/porous layer open tubular/mass spectrometry for ultratrace proteomic analysis using a 10 microm id poly(styrene- divinylbenzene) porous layer open tubular column with an on-line triphasic trapping column. AB - This study expands the capabilities for ultratrace proteomic analysis of our previous work by incorporating on-line sample desalting using a triphasic (RP/strong cation exchange (SCX)/micro-SPE) trapping column connected to a 3.2 m x 10 microm id poly(styrene-divinylbenzene) (PS-DVB) porous layer open tubular (PLOT) column. To minimize extra sample handling steps, C18 RP packing was incorporated in the capillary tubing upstream of the SCX column for the on-line desalting. For the micro-SPE column, a 50 microm id PS-DVB monolithic column was positioned downstream of the SCX column. High-performance separation was achieved on the PLOT column at a mobile phase flow rate of 20 nL/min. The sensitivity and high resolution capability of the new multidimensional platform was evaluated using an in-gel tryptic digested sample of a cervical cancer (SiHa) cell line. For the injected amount of 1200 cells ( approximately 500 ng), over 2700 peptides covering greater than 850 unique proteins were identified from the triphasic SCX/PLOT/MS analysis of a single SDS gel section (>40 kDa). The 2-D LC/MS platform demonstrated good separation performance, such that more than 85% of the identified peptides were detected from only one salt fraction. In a triplicate analysis of the above >40 kDa gel section, 4497 peptides and 1209 unique proteins were identified when applying stringent filtering criteria, with a false-positive rate of 2.4%. When all three SDS-PAGE gel sections of the lysed SiHa cells were analyzed, 5047 peptides and 1857 unique proteins (false-positive rate 1.8%), including cancer-related proteins such as MAP kinases, were identified. PMID- 18383018 TI - High-resolution computer simulation of electrophoretic mobilization in isoelectric focusing. AB - Cationic and anionic electrophoretic mobilization for focusing of hemoglobins (Hb's) in the presence of 100 carrier ampholytes covering a pI range of 6.00-7.98 was studied by computer simulation at a constant current density of 300 A/m(2). Electropherograms that would be produced by whole column imaging and by single detectors placed at different locations along the focusing column are presented. Upon mobilization, peak heights of the Hb zones decrease, but the zones retain a relatively sharp constant profile and are migrating at a constant velocity. A further peak decrease occurs during readjustment at the locations of the original buffer/column interfaces, indicating that detection sensitivity is the lowest at these locations. An anionic carrier ampholyte mobility smaller than that of its cationic species produces a cathodic drift which is smaller than the transport rate used for electrophoretic mobilization. Compared to the case with equal mobilities of carrier ampholyte species, a small increase (decrease) is predicted for the cationic (anionic) mobilization rate within the focusing column. Simulation data suggest that electrophoretic mobilization after focusing and focusing with concurrent electrophoretic mobilization are comparable isotachophoretic processes that occur when there is an uninterrupted flux of an ion through the focusing column. Cathodic drift caused by unequal mobilities of the species of carrier ampholytes, electrophoretic mobilization, and decomposition occurring at the pH gradient edges are related electrophoretic processes. PMID- 18383019 TI - Capillary electrophoresis of boron cluster compounds in aqueous and nonaqueous solvents. AB - The electrophoretic properties of boron cluster compounds were determined in water, methanol and ACN as solvents of the BGE and discussed based on the principles of ion migration. Two types of boron cluster compounds were investigated. One type consisted of derivatives of the nido-7,8 dicarbaundecaborate cluster, the other types are derivatized cobalt bis(dicarbollide) ions (COSANs) whose central cobalt atom is sandwiched by two 7,8-dicarbaundecaborate clusters. The BGE in all solvents was acetate/acetic acid buffer with pH 4.75 in water, 9.7 in methanol and 22.3 in ACN, respectively, at different ionic strength between 5 and 30 mM. The dependence of the mobility on ionic strength could not be explained by the theory of Debye, Huckel and Onsager, but good agreement was found upon considering an ion size parameter. Limiting mobilities were derived by curve fitting, and by the aid of the solvent viscosities the hydrodynamic radii of the analyte anions were calculated. They are between 0.25 and 0.48 nm, and were nearly independent of the solvent. Electrophoresis of the analytes in a BGE consisting of 6 mM perchloric acid in ACN allows the conclusion that the present boron cluster compounds behave as stronger acids than perchloric acid. PMID- 18383020 TI - Coupling of solid-phase microextraction continuous bed (monolithic) capillaries with capillary zone electrophoresis for direct analysis of drugs in biological fluids. AB - Hyperlink robust biocompatible solid-phase microextraction (SPME) devices were prepared using continuous bed (monolithic) restricted-access media (RAM) as the SPME capillary insert. The RAM-based SPME approach was able to simultaneously separate proteins from a biological sample, while directly extracting the active components of caffeine, paracetamol and acetylsalicylic acid from the drug NeoCitramonum. The devices were interfaced with a CZE system and fully automated analysis for sample preconcentration, desorption, separation and quantification of analytes was evaluated. Comparative study of in-line coupled SPME-CZE using RAM and RP capillary inserts was carried out. Using an SPME (RAM) insert, the calculated caffeine, paracetamol and acetylsalicylic acid LODs in a bovine plasma sample were 0.3, 0.8 and 1.9 ng/mL, respectively. PMID- 18383021 TI - Insulin-like growth factor-induced signals activate mitochondrial respiration. AB - From experiments with lower eukaryotes it is known that the metabolic rate and also the rate of aging are tightly controlled by the insulin-like growth factor (IGF)/insulin signal transduction pathway. The mitochondrial theory of aging implies that an increased metabolic rate leads to increased mitochondrial activity; increased production of reactive oxygen species due to these alterations would speed up the aging process. To address the question if mitochondrial activity is influenced by insulin/IGF signaling, we have established an experimental system to determine the influence of IGF-I-dependent signaling on mitochondrial function. We used DU145 prostate cancer cells, known for the intact IGF signal transduction pathway, to address the influence of IGF receptor activation on mitochondrial function by high-resolution respirometry. These experiments revealed that indeed mitochondrial function is regulated by IGF signaling, and up-regulation of respiration seems to require phosphoinositide 3 kinase/AKT signaling, but is independent of IGF effects on cell cycle progression. Collectively these data establish a regulatory cross-talk between insulin/IGF signal transduction and mitochondrial function, two major pathways implicated in controlling the rate of aging. PMID- 18383022 TI - Cell-based assays in GPCR drug discovery. AB - G protein-coupled receptors (GPCRs) transmit extracellular signals into the intracellular space, and play key roles in the physiological regulation of virtually every cell and tissue. Characteristic for the GPCR superfamily of cell surface receptors are their seven transmembrane-spanning alpha-helices, an extracellular N terminus and intracellular C-terminal tail. Besides transmission of extracellular signals, their activity is modulated by cellular signals in an auto- or transregulatory fashion. The molecular complexity of GPCRs and their regulated signaling networks triggered the interest in academic research groups to explore them further, and their drugability and role in pathophysiology triggers pharmaceutical research towards small molecular weight ligands and therapeutic antibodies. About 30% of marketed drugs target GPCRs, which underlines the importance of this target class. This review describes current and emerging cellular assays for the ligand discovery of GPCRs. PMID- 18383023 TI - Dietary restriction by growth in axenic medium induces discrete changes in the transcriptional output of genes involved in energy metabolism in Caenorhabditis elegans. AB - Dietary restriction increases life span in a wide range of species, including the nematode worm Caenorhabditis elegans. The mechanism by which it does so remains largely unknown, although it is commonly thought that a reduction of reactive oxygen species (ROS) plays a pivotal role. More specifically, for C. elegans, it has been proposed that food restriction reduces energy expenditure, possibly in conjunction with an anaerobic shift in energy production, with consequent reduction in the formation of ROS. We have measured differential transcript abundance of 49 genes known to play roles in energy metabolism in axenic culture medium, which causes a nutritional deficit and leads to a substantial increase of life span. We found no evidence for a reduction in metabolic rate or a shift to anaerobic metabolism in axenic culture. Major changes induced by growth in axenic medium include down-regulation of lipid degradation and up-regulation of glyoxylate cycle activity glyceroneogenesis and, possibly, gluconeogenesis. The activities determined in worm extracts for pyruvate kinase, phosphoenolpyruvate carboxykinase and isocitrate lyase followed a similar trend. We conclude that growth in axenic culture is marked by a general up-regulation of replenishing pathways. PMID- 18383024 TI - Direct injection of organic solvent extracts for capillary electrophoresis. AB - It is demonstrated here that organic solvents immiscible in water used in sample extraction, such as chloroform, can be injected directly and successfully on the capillary without the need for evaporation and reconstitution. Current continuity was maintained all the time during the run. In order to avoid the rapid evaporation of the organic solvent during the analysis, the aqueous layer was left over the chloroform. This simplified the extraction step, and enabled the injection from the same vial over several hours without dealing with problem of evaporation. The relative peak heights in the electropherograms can be modified by the inclusion in the chloroform of a more polar solvent, by adjusting the pH, or adjusting the salt content of the sample. Addition of a polar solvent to the chloroform improved greatly the precision of the analysis for both the peak height and migration time. PMID- 18383025 TI - Electrochromatography in microchips packed with conventional reversed-phase silica particles. AB - This paper shows the applicability of a disposable and inexpensive microfluidic chip for electrochromatographic separations. The chip, recently developed by us for chip-based LC, was fabricated from PDMS incorporating conventional chromatographic RP silica particles (C18) without the use of frits. Three cephalosporin antibiotics were used to demonstrate the applicability of the chip based chromatographic packing for electrochromatographic determinations. The used sample injection method utilizes hydrodynamic pressure, thereby, reducing the propensity for sample bias during the injection. PMID- 18383026 TI - The influence of Cu(2+) on the unfolding and refolding of intact and proteolytically processed beta(2)-microglobulin. AB - Human beta(2)-microglobulin (beta(2)m) is an amyloidogenic protein in patients suffering from chronic kidney disease and especially in those patients that need intermittent hemodialysis for longer periods, e.g., when awaiting transplantation. While many in vitro conditions induce beta(2)m-amyloid formation from wild-type (wt) beta(2)m and while a number of structurally altered beta(2)m molecules are known to be conformationally unstable and amyloidogenic on their own, it is not known why beta(2)m-amyloid is generated in some dialysis patients. For many amyloid proteins it is known that divalent metal ions, especially Cu(2+), display strong binding and distinct destabilizing effects on protein conformation. The present study uses CE to assess conformational states of wt and cleaved beta(2)m (dK58-beta(2)m, beta(2)m cleaved at lysine-58, a modification found in the circulation of hemodialysis patients) in the presence of divalent metal ions. The experiments provide both qualitative and quantitative data showing the specific destabilizing effects of Cu(2+)-ions on the folding of wt beta(2)m. Both refolding after acid denaturation and solution structure of beta(2)m under otherwise native conditions are severely influenced by Cu(2+). An increased unfolding, aggregation, and induction of Congo red-reactive molecular species in Cu(2+)-incubated wt-beta(2)m could be demonstrated while the refolding kinetics of dK58-beta(2)m, already slower than the wt molecule, appeared not to be further decreased by Cu(2+). Given the interest in the actions of metal ions in other types of amyloidosis, including, e.g., Alzheimer's disease and the prion encephalopathies, the use of microelectrophoretic methods to monitor unfolding and refolding of biomolecules available in scarce amounts as shown in this study is an attractive option. PMID- 18383027 TI - Automation of nanoflow liquid chromatography-tandem mass spectrometry for proteome and peptide profiling analysis by using a monolithic analytical capillary column. AB - An automated nano-LC-MS/MS platform without trap column was established, which only used a 20 cm lauryl methacrylate-ethylene dimethacrylate (LMA-EDMA) monolithic capillary column to allow preconcentration and separation of peptides. The monolithic column had the advantages of good permeability and low backpressure resulting in higher flow rates for capillary columns. Tryptic digests of bovine albumin and yeast protein extract were tested using the monolithic column system. High proteomic coverage using this approach were demonstrated in this study. Furthermore, peptide samples extracted from mouse liver were separated by using the monolithic column system combined with size exclusion chromatography prefractionation. This monolithic column system might be a promising alternative for the automated system previously using a trap column for routine proteome and peptide profiling analysis. PMID- 18383028 TI - Enantioselective silica-based monoliths modified with a novel aminosulfonic acid derived strong cation exchanger for electrically driven and pressure-driven capillary chromatography. AB - Silica monoliths modified with trans-(1S,2S)-2-(N-4-allyloxy-3,5 dichlorobenzoyl)amino cyclohexanesulfonic acid were tested for enantioselective separations of various chiral bases by aqueous and nonaqueous CEC as well as nano HPLC. The optimization of the immobilization procedure showed that an intermediate selector (SO) coverage, as does result from a single static immobilization cycle in the capillary at 60 degrees C with an 8% (m/v) SO solution in methanol, affords maximal EOF and optimal enantioselectivity values, while a second immobilization cycle does not lead to any improvements. Furthermore, the mobile phase composition was examined regarding the effectiveness of aqueous phases (ACN/water and methanol/water) compared to nonaqueous eluents (ACN/methanol) in terms of separation selectivity and efficiency. Additionally, different acids of varying strengths were tested as co ions in the ion-exchange process, including formic acid, acetic acid, methanesulfonic acid, and TFA (pK(a) from 4.75 to 0.5). It turned out that the effects regarding EOF and enantioselectivity were largely negligible. The chromatographic efficiencies of the new capillary columns were compelling and remarkable for bases. H-u curves established for mefloquine revealed a C-term contribution (resistance to mass transfer) by a factor of about six lower in CEC than in nano-HPLC and an A-term (flow maldistribution) about three times lower in the CEC mode. Theoretical plate heights as low as around 3-5 mum could be obtained in CEC over a wide flow range (0.5-1.5 mm/s). Run-to-run repeatabilities like in HPLC and excellent system stability promise the practical usefulness of the novel monolithic capillary column for enantiomeric composition analysis of pharmaceuticals by CEC. PMID- 18383029 TI - Capillary electrophoretic separation of biologically active amines and acids using nanoparticle-coated capillaries. AB - This manuscript describes dynamic coating of capillaries with poly(L-lysine) (PLL) and silica nanoparticles (SiO2 NPs) and use of the as-prepared capillaries for the separation of biogenic amines and acids by CE in conjunction with LIF detection. The directions of EOF are controlled by varying the outmost layer of the capillaries with PLL and SiO2 NPs, respectively. Over the pH range 3.0-5.0, the (PLL-SiO2NP)n-PLL capillaries have an EOF toward the anodic end and are more suitable for the separation of acids with respect to speed, while the (PLL SiO2NP)n capillaries have an EOF toward the cathodic end and are more suitable for the separation of biogenic amines regarding speed and sensitivity. The separations of standard solutions containing five amines and two acids by CE with LIF detection using (PLL-SiO2NP)2-PLL and (PLL-SiO2NP)3 capillaries were accomplished within 10 and 7 min, providing plate numbers of 3.8 and 5.0x10(4) plates/m for 5-hydroxytryptamine (5-HT), respectively. The LODs for 5-HT and 5 hydroxyindole-3-acetic acid (5-HIAA) are 32 and 2 nM and 0.2 and 1.5 nM when using the (PLL-SiO2NP)2-PLL and (PLL-SiO2NP)3 capillaries, respectively. Identification and quantification of 5-HIAA, homovanillic acid, and DL vanillomandelic acid in urine samples from a male before and after drinking green tea were tested to validate practicality of the present approach. The results show that the (PLL-SiO2NP)2-PLL capillary provides greater resolving power, while the (PLL-SiO2NP)3 capillary provides better sensitivity, higher efficiency, and longer durability for the separation of the amines and acids. PMID- 18383031 TI - Nonaqueous capillary electrophoresis for analysis of the ethanol consumption biomarker phosphatidylethanol. AB - Nonaqueous CE (NACE) methodology was developed for the separation and determination of phosphatidylethanol (Peth), a new biomarker of ethanol intake. Peth is an abnormal phospholipid formed in cell membranes only in the presence of ethanol, via the transphosphatidylation reaction of phospholipase D. The NACE separation medium consisted of 80 mM ammonium acetate in 50% ACN, 33% 2-propanol, 12% hexane and 5% methanol. A stacking effect was obtained by reducing the concentration of ammonium acetate in the separation medium for all injected samples. The LOD was estimated to 1 microM (5.6 fmol) of Peth with conventional UV detection, equalling 0.4 micromol/L blood. Peth was successfully determined in extracts of human blood samples. Separation of Peth from other blood lipids in the lipid extract sample was performed in 5 min. The method facilitates smaller sample volumes and performs about ten times faster compared to earlier chromatographical methods. PMID- 18383030 TI - Analysis of phenolic compounds in extra virgin olive oil by using reversed-phase capillary electrochromatography. AB - In this work, the simultaneous separation of ten phenolic compounds (protocatechuic, p-coumaric, o-coumaric, vanillic, ferulic, caffeic, syringic acids, hydroxytyrosol, tyrosol and oleuropein) in extra virgin olive oils (EVOOs) by isocratic RP CEC is proposed. A CEC method was optimized in order to completely resolve all the analyzed compounds by studying several experimental parameters. The influence of the stationary phase type (C(18) and C(8) modified silica gel), buffer concentration and pH as well as the organic modifier content of the mobile phase on retention factors, selectivity and efficiency were evaluated in details. A capillary column packed with Cogent bidentate C(18) particles for 23 cm and a mobile phase composed by 100 mM ammonium formate buffer pH 3/H(2)O/ACN (5:65:30 v/v/v) allowed the baseline resolution of the compounds under study in less than 35 min setting the applied voltage and temperature at 22 kV and 20 degrees C, respectively. A study, evaluating the intra- and interday precision as well as LOD and LOQ and method linearity was developed in accordance with the analytical procedures for method validation. LODs were in the range of 0.015-2.5 microg/mL, while calibration curves showed a good linearity (r(2) >0.997). The CEC method was applied to the separation and determination of these compounds in EVOO samples after a suitable liquid-liquid extraction procedure. The mean recovery values of the studied compounds ranged between 87 and 99%. PMID- 18383032 TI - Capillary electrochromatographic separation of peptides using a macrocyclic polyamine for molecular recognition. AB - A macrocyclic polyamine, 1,5,9,13,17,21,25,29-octaazacyclodotriacontane ([32]ane N(8)), in the bonded phase was employed as a molecular receptor for CEC separation of oligopeptides. Parameters affecting the performance of the separations were considered. Baseline separation for the mixture of angiotensin I, angiotensin II, [Sar(1), Thr(8)]-angiotensin II, beta-casomorphin bovine, beta casomorphin human, oxytocin acetate, tocinoic acid, vasopressin, and FMRF amide could be achieved using phosphate buffer (30 mM, pH 7) as the mobile phase. Column efficiency with average theoretical plate numbers of 69 000 plates/m and RSDs of <1% (n = 6) was achieved. [Met(5)]-enkephalin and [Leu(5)]-enkephalin, which have identical pI values and similar masses could be completely separated using acetate buffer (30 mM) with pH gradient (pH 3 inlet side and pH 4 outlet side). The results suggest that the mechanism for the peptide separation was mediated by a combination of electrophoretic migration and chromatographic retention involving anion coordination and anion exchange. After long-term use, the deviation of the EOF of the column after more than 600 injections was still within 6.0% of that for a freshly prepared column. PMID- 18383033 TI - Stellan Hjerten's contribution to the development of monolithic stationary phases. AB - This overview is presented to celebrate the birthday of one of the luminaries of the separation science and my friend - Stellan Hjerten. He made significant contributions to a variety of areas in separation science such as electrophoresis, LC, and CEC to name just a few. Since the scope of his work was enormous, this review will focus only on a single aspect of his scientific activities, the design and applications of monolithic materials. During the years starting from 1989, Stellan Hjerten published many excellent papers concerning the preparation of acrylamide chemistry-based monoliths and their use in both micro-HPLC and CEC. The following text details his works in the field. PMID- 18383034 TI - Th17 development and autoimmune arthritis in the absence of reactive oxygen species. AB - Dendritic cells (DC) express a functional NADPH oxidase and produce reactive oxygen species (ROS) upon interaction with microbes and T cells. Exposure to ROS leads to DC activation and maturation, as evidenced by phenotypic and functional changes. We have evaluated how endogenous ROS production affects the cytokine secretion pattern and T cell-activating capacity of bone marrow-derived murine DC. DC treated with ROS scavengers, as well as DC from mice that lack a functional NADPH oxidase (and thereby inherently deficient in ROS production) produced significantly increased levels of IL-1beta, IL-6, TNF-alpha and TGF-beta in response to microbial activation. DC deficient in ROS production induced high levels of IFN-gamma and IL-17 in responding T cells after Ag-specific or superantigen-induced activation. Finally, we show that ROS deficiency affected the induction of a T cell-dependent inflammatory condition, collagen-induced arthritis (CIA). C57BL/6 mice that lack a functional NADPH oxidase developed a severe and erosive CD4-dependent CIA, whereas the majority of the congenic wild type animals remained healthy. These data suggest that ROS act as immunomodulators in DC-driven T cell activation and perhaps also in T cell dependent immunopathology. PMID- 18383035 TI - A novel form of NF-kappaB is induced by Leishmania infection: involvement in macrophage gene expression. AB - Leishmania spp. are obligate intracellular parasites that inhabit the phagolysosomes of macrophages. Manipulation of host cell signaling pathways and gene expression by Leishmania is critical for Leishmania's survival and resultant pathology. Here, we show that infection of macrophages with Leishmania promastigotes in vitro causes specific cleavage of the NF-kappaB p65 RelA subunit. Cleavage occurs in the cytoplasm and is dependent on the Leishmania protease gp63. The resulting fragment, p35 RelA, migrates to the nucleus, where it binds DNA as a heterodimer with NF-kappaB p50. Importantly, induction of chemokine gene expression (MIP-2/CXCL2, MCP-1/CCL2, MIP-1alpha/CCL3, MIP 1beta/CCL4) by Leishmania is NF-kappaB dependent, which implies that p35 RelA/p50 dimers are able to activate transcription, despite the absence of a recognized transcriptional transactivation domain. NF-kappaB cleavage was observed following infection with a range of pathogenic species, including L. donovani, L. major, L. mexicana, and L. (Viannia) braziliensis, but not the non-pathogenic L. tarentolae or treatment with IFN-gamma. These results indicate a novel mechanism by which a pathogen can subvert a macrophage's regulatory pathways to alter NF-kappaB activity. PMID- 18383037 TI - Notch signaling is necessary for GATA3 function in the initiation of T cell development. AB - GATA3 and Notch1 are essential for T cell development at the earliest stage, but their mutual roles in this process remain to be clarified. In this study, we demonstrated that impairment of T lymphopoiesis in hematopoietic progenitor cells (HPC) of GATA3-deficient fetal liver (FL) on day 11.5 of gestation (E11.5) was rescued only by introduction of both GATA3 and the intracellular region of Notch1 but not by either alone. However, the introduction of GATA3 only was sufficient for T cell induction in GATA3-deficient FL cells at the advanced stage, where Notch signaling is well detectable. This indicates that Notch signaling is necessary for GATA3 to function for T cell fate specification but is not sufficient without GATA3. On the other hand, Notch signaling is sufficient for blockage of B cell development without GATA3, suggesting that T cell fate specification at the branching point does not result simply from the developmental arrest of B cell lineage by Notch signaling. PMID- 18383036 TI - Proliferation and interleukin 5 production by CD8hi CD57+ T cells. AB - CD8hi CD57+ T cells have previously been described as effector memory T cells with minimal expansion capacity and high susceptibility to activation-induced cell death. In contrast, we demonstrate here that CD8hi CD57+ T cells are capable of rapid expansion using multiple techniques including [(3)H]thymidine uptake, flow cytometric bead-based enumeration and standard haemocytometer counting. Previous reports can be explained by marked inhibition of activation-induced expansion and increased 7-amino-actinomycin D uptake by CD8hi CD57+ T cells following treatment with CFSE, a dye previously used to measure their proliferation, combined with specific media requirements for the growth of this cell subset. The ability of CD8hi CD57+ T cells to further differentiate is highlighted by a distinct cytokine profile late after activation that includes the unexpected release of high levels of interleukin 5. These data indicate that CD8hi CD57+ T cells should not be considered as "end-stage" effector T cells incapable of proliferation, but represent a highly differentiated subset capable of rapid division and exhibiting novel functions separate from their previously described cytotoxic and IFN-gamma responses. PMID- 18383038 TI - Preservation of a critical epitope core region is associated with the high degree of flaviviral cross-reactivity exhibited by a dengue-specific CD4+ T cell clone. AB - Dengue is a member of the Flaviviridae, a large group of related viruses some of which co-circulate in certain regions (e.g. dengue and Yellow fever in South America). Immune responses cross-reactive between different dengue serotypes are important in the pathogenesis of dengue disease but it is not known whether previous infection with one flavivirus might affect the clinical course of subsequent infections with other members of the family. CD4+ T cells have been shown to be important in the production of cytokines in response to dengue infection and can demonstrate significant epitope cross-reactivity. Here, we describe the generation and characterisation of CD4+ T cell clones from a patient experiencing acute dengue infection. These clones were DRB1*15+ and recognised epitope variants not only within other dengue viruses but certain other flaviviruses. This cross-reactivity was dependent upon the presence of a five amino acid core region, consistent with structural observations of class II MHC binding to TCR demonstrating that only a subset of residues within an epitope bound to a class II molecule are "read out" by the TCR. This capacity of CD4+ T cell clones to recognise a given epitope despite considerable variation between viruses may be of pathological significance, particularly in regions where related viruses co-circulate. PMID- 18383039 TI - Chimpanzee CD4+ T cells are relatively insensitive to HIV-1 envelope-mediated inhibition of CD154 up-regulation. AB - CD40-CD154 interaction forms a key event in regulation of crosstalk between dendritic cells and CD4 T cells. In human immunodeficiency virus (HIV)-1 infected patients CD154 expression is impaired, and the resulting loss of immune responsiveness by CD4+ T cells contributes to a progressive state of immunodeficiency in humans. Although chimpanzees are susceptible to chronic HIV 1/SIVcpz infection, they are relatively resistant to the onset of AIDS. This relative resistance is characterized by maintenance of CD4+ T cell populations and function, which is highly compromised in human patients. In our cohort of chronically HIV-1- and SIVcpz-infected chimpanzees, we demonstrated the capacity to produce IL-2, following CD3/CD28 stimulation, as well as preserved CD154 up regulation. Cross-linking of CD4 with mAb was found to inhibit CD3/CD28-induced up-regulation of CD154 equally in chimpanzees and humans. However, specific cross linking with trimeric recombinant HIV-1 gp140 revealed reduced sensitivity for inhibition of CD154 up-regulation in chimpanzees, requiring fourfold higher concentrations of viral protein. Chimpanzee CD4+ T cells are thus less sensitive to the immune-suppressive effect of low-dose HIV-1 envelope protein than human CD4+ T cells. PMID- 18383040 TI - Interleukin-10 regulates TNF-alpha-converting enzyme (TACE/ADAM-17) involving a TIMP-3 dependent and independent mechanism. AB - IL-10 is a potent anti-inflammatory molecule, which regulates TNF-alpha at multiple levels. We investigated whether IL-10 also modulated the activity of the TNF-alpha-converting enzyme (TACE). Using an ex vivo fluorogenic assay we observed that LPS rapidly induced TACE activity in monocytes coinciding with release of soluble TNF-alpha. In the presence of IL-10, TNF-alpha production and activation of surface TACE was significantly inhibited. Paradoxically, both LPS with or without IL-10 led to accumulation of surface TACE (albeit catalytically inactive) over a 24 h period. We investigated whether this was mediated through induction of endogenous tissue inhibitor metalloproteinase-3 (TIMP-3). We found that the inhibition of TACE activity at 2 h by IL-10 was not TIMP-3 dependent but that the late accumulation of surface TACE was prevented with TIMP-3 antibodies. Furthermore, induction of endogenous TIMP-3 was observed by western blotting in both LPS- and in LPS with IL-10-treated monocytes from 6 to 8 h of culture. These results indicate that IL-10 further regulates TNF-alpha by modulating TACE activation at early time points and by contributing to the induction of TIMP-3, the natural inhibitor of active TACE, at later time points. These observations add to our understanding of inflammation and the importance of homeostatic regulators of these events. PMID- 18383041 TI - Defective positive selection results in T cell lymphopenia and increased autoimmune diabetes in ADAP-deficient BDC2.5-C57BL/6 mice. AB - Adhesion and degranulation promoting adapter protein (ADAP), a positive regulator of T cell receptor (TCR) signaling, is required for thymocyte development and T cell homeostasis. To investigate the role of ADAP in a T cell-driven autoimmune response, we generated ADAP-deficient, BDC2.5 TCR transgenic, diabetes-prone (C57BL/6) mice (BDC/B6). We observed a striking enhancement of diabetes incidence in ADAP-deficient mice, both in animals homozygous for I-Ag7, and in mice carrying one I-Ab allele (BDC/B6g7/b). Increased disease correlates with significantly reduced numbers of pathological CD4(+) T cells in the mice. Consistent with a state of functional lymphopenia in ADAP-deficient BDC/B6g7/b mice, T cells display increased homeostatic proliferation. Transfer of syngeneic lymphocytes or T cells both blocks ADAP-dependent diabetes and relieves exaggerated homeostatic T cell proliferation observed in ADAP-deficient mice. Marked attenuation in cellularity of the CD4+ single-positive thymocyte compartment in ADAP-deficient BDC/B6g7/b animals suggests a mechanism for induction of the lymphopenia. We conclude that inefficient positive selection in ADAP deficiency results in lymphopenia that leads to enhanced autoimmune diabetes in the BDC/B6g7/b model. Our findings support the notion that ineffective thymic T cell output can be a powerful causative factor in lymphopenia-driven autoimmune diabetes. PMID- 18383042 TI - CXCR3 determines strain susceptibility to murine cerebral malaria by mediating T lymphocyte migration toward IFN-gamma-induced chemokines. AB - Cerebral malaria (CM) results from the binding of infected erythrocytes and leukocytes to brain endothelia. The precise mechanisms underlying lymphocyte recruitment and activation in CM remain unclear. Therefore, the expression of various chemokines was quantified in brains of mice infected with Plasmodium berghei ANKA (PbA). Several chemokines attracting monocytes and activated T lymphocytes were expressed at high levels. Their expression was almost completely abrogated in IFN-gamma ligand and receptor KO mice, indicating that IFN-gamma is an essential chemokine inducer in vivo. Surprisingly, the expression levels of chemokines, IFN-gamma and also adhesion molecules in the brain were not lower in CM-resistant Balb/c and DBA/2 mice compared to CM-sensitive C57BL/6 and DBA/1 mice, although T lymphocyte sequestration in the brain was significantly less in CM-resistant than in CM-sensitive mice. This difference correlated with a higher up-regulation of the CXC chemokine receptor (CXCR)-3 on splenic T cells and a higher chemotactic response to IFN-gamma-inducible protein-10 (IP-10) in C57BL/6 compared to Balb/c mice. In conclusion, parasite-induced IFN-gamma in the brain results in high local expression levels of specific chemokines for monocytes and lymphocytes. The strain-dependent susceptibility to develop CM is more related to the expression of CXCR3 in circulating leukocytes than to the chemokine expression levels in the brain. PMID- 18383043 TI - Liver sinusoidal endothelial cells veto CD8 T cell activation by antigen presenting dendritic cells. AB - The liver is known to induce tolerance rather than immunity through tolerogenic antigen presentation or elimination of effector T cells. In particular, hepatic dendritic cells (DC) are known to be little immunogenic for CD8 T cells. Here, we investigated whether this peculiar phenotype resulted from interaction with resident hepatic cell populations. Contact of DC with liver sinusoidal endothelial cells (LSEC) but not hepatocytes or B cells vetoed antigen-presenting DC to fully activate naive CD8 T cells. This MHC-independent regulatory effect of LSEC on DC function was not connected to soluble mediators but required physical contact. Because interaction with third-party LSEC still allowed antigen presenting DC to stimulate expression of initial activation markers on naive CD8 T cells and to stimulate activated CD8 T cells, we hypothesize that LSEC controlled the DC costimulatory function. Indeed, contact with LSEC led to reduced DC expression levels of CD80/86 or IL-12, but supplementation of these signals failed to rescue the ability to prime naive CD8 T cells, indicating involvement of further molecules. Taken together, our results reveal a novel principle operative in hepatic tolerance induction, in which LSEC not only tolerize T cells themselves but also suppress neighboring APC normally capable of inducing T cell immunity. PMID- 18383044 TI - Will genetic profiling make a difference in clinical trial outcomes? AB - What stimulated me to write this editorial was an article I read in USA Today written by Lois Hatton, who is a writer and columnist based in Brookings, South Dakota and author of "Inspiration for a Lifetime." The article, "'Black' Cherokees fight for heritage," told of a black Yankton Sioux who says he is seen as a black by other Sioux while on the reservation, but as an Indian by blacks who do not live on the reservation. PMID- 18383045 TI - Systolic function as a predictor of mortality and quality of life in long-term survivors with heart failure. AB - BACKGROUND: Patients with heart failure (HF) and preserved ejection fraction (EF) have been shown to have high mortality rates, comparable to those with reduced EF. Thus, long-term survivors of HF, regardless of ejection fraction, are a select group. Little is known about disease-related quality of life (QOL) and health status in these patients. HYPOTHESIS: Preserved EF in patients with heart failure independently predicts long-term survival, health related quality of life (QOL), or functional status. METHODS: The study followed a cohort of 413 patients consecutively hospitalized for HF between March 1996 and September 1998. In July 2005, information was collected about their mortality, health related QOL as defined by disease-specific Kansas City Cardiomyopathy Questionnaire (KCCQ) scores, and functional decline as defined by limitations in Activities of Daily Living (ADL) scores. RESULTS: The primary outcomes were mortality, QOL, and functional decline. At follow-up, 8.1 years after enrollment, overall mortality was 76% (314/413). Adjusted for age, gender, renal insufficiency, diabetes mellitus, hypertension, HF, and respiratory disease, those with decreased ejection fraction (EF < 40%) had higher mortality compared with those with preserved ejection fraction (hazard ratio [HR] 1.42; confidence interval [CI] = 1.13, 1.80, p = 0.003). The KCCQ scores, including Clinical Summary Scores and Symptom Limitation Scores, as well as ADL limitations, were not significantly different in the survivors with preserved or decreased EF. CONCLUSIONS: Heart failure patients with preserved EF have a modest survival advantage compared with those with decreased EF, but health related QOL scores and functional decline in survivors are similar regardless of systolic function. PMID- 18383046 TI - Profiles in cardiology. W. Bruce Fye. PMID- 18383047 TI - Wellens' syndrome or inverted U-waves? AB - This report describes an example of a patient with chest pain and classic electrocardiographic changes consistent with ischemia in the proximal left anterior descending coronary artery (LAD) territory. This electrocardiogram (ECG) abnormality is known as Wellens' syndrome. However, the same abnormality is also described as inverted U-waves. The significance of this abnormality and the controversy of describing this syndrome is reviewed and discussed. PMID- 18383048 TI - Worse prognosis with gene mutations of beta-myosin heavy chain than myosin binding protein C in Chinese patients with hypertrophic cardiomyopathy. AB - BACKGROUND: No data are available on survival analysis and longitudinal evolution of patients with gene mutations of beta-myosin heavy chain (MYH7) and myosin binding protein C (MYBPC3) in Chinese. HYPOTHESIS: To prospectively investigate whether different gene mutations confer distinct prognosis. METHODS: We performed a prospective study in 70 HCM patients and 46 genetically affected family members without HCM-phenotype with direct DNA sequencing of MYH7 and MYBPC3, clinical assessments, and 5.8 +/- 1.8 years follow-up. RESULTS: After follow-up, more surgical intervention (8/52 versus 0/18, p < 0.001), higher sudden death risk (7/52 versus 0/18, p < 0.001) and shorter life span were found in patients with MYH7 mutations than in patients with MYBPC3 mutations (45.1 +/- 14.0 versus 73.5 +/- 7.5 years, p = 0.03). Seven of the 27 mutation carriers of MYH7 had clinical presentations of HCM, but no carriers of MYBPC3 mutations developed to HCM during follow-up. Maximal wall thickness was thicker in the patients carrying mutations in the global region of MYH7 than in those carrying mutations in the rod region of MYH7 (21.5 +/- 6.6 versus 15 +/- 6.1 mm, p < 0.05) at baseline. More sudden death (7/41 versus 0/11) and left ventricular dysfunction (NYHA Class III approximately IV, 17/32 versus 1/10) were identified in patients with mutations in the global region of MYH7 than in patients with other mutations. CONCLUSIONS: MYH7 mutations, especially in the global region, cause malignant clinical phenotypes. PMID- 18383049 TI - Prognostic value of troponins in patients with non-ST-segment elevation acute coronary syndromes and chronic kidney disease. AB - BACKGROUND: The prognostic value of cardiac troponins (cTn) in patients with non ST-segment elevation acute coronary syndromes (NSTE ACS) and chronic kidney disease (CKD) is debated. HYPOTHESIS: We tested the performance of cTnI and cTnT for risk stratification in patients with CKD and evaluated the prognostic significance of cTnI and cTnT elevations by their magnitude across the range of CKD severity. METHODS: We examined correlations among cTn elevation, CKD, and in hospital mortality in 31,586 high-risk patients with NSTE ACS included in the Can Rapid Risk Stratification of Unstable Angina Patients Suppress ADverse Outcomes with Early Implementation of the American College of Cardiology/American Heart Association Guidelines initiative (CRUSADE). Cardiac tropinins I and T levels were categorized as ratios of each site's upper limit of normal (ULN) for myocardial necrosis: normal (cTn ratio < or =1 x ULN), mild (cTn ratio > 1-3 x ULN), and major (cTn ratio > 3 x ULN) elevation. Estimated glomerular filtration rate (eGFR) was calculated using the abbreviated Modification of Diet in Renal Disease equation. Stages of CKD were categorized as normal to mild (eGFR > 60 mL/min), moderate (eGFR 30-60 mL/min), or severe (eGFR < 30 mL/min). RESULTS: Mortality increased more steeply across CKD stages (2.0%-12.9%) than across cTn ratio categories (2.7%-5.4%). In normal or mild CKD, mortality was low regardless of cTn elevations. In moderate CKD, mortality increased incrementally with cTnI (3.3% versus 5.4% versus 7.4%) and cTnT (3.7% versus 5.3% versus 7.3%) elevation. Among severe CKD patients, only major cTn elevations further distinguished risk (cTnI: 10.1% versus 9.7% versus 14.6%; cTnT: 7.0% versus 5.7% versus 14.0%). CONCLUSIONS: In patients with CKD, cTnI and cTnT perform equally in differentiating short-term prognosis following NSTE ACS; however, the prognostic impact of cTn is dependent upon the degree of CKD severity. PMID- 18383050 TI - Update on atrial fibrillation: part II. AB - Antiarrhythmic drugs are an essential tool in the management of atrial fibrillation (AF). Although we are already on the threshold of a large expansion in the use of ablation therapies, these will not, however, be appropriate for all patients, and pharmacological therapies will continue to have an important place in the management of atrial fibrillation. The plethora of antiarrhythmic drugs currently available for the treatment of atrial fibrillation is a reflection that none is wholly satisfactory, each having limited efficacy combined with poor safety and tolerability. Improved class III antiarrhythmic drugs, such as dronedarone, new classes of antiarrhythmic agents, such as atrial repolarization delaying agents, and upstream therapies dealing with substrate, represent potential sources of new pharmacological therapies. PMID- 18383051 TI - Postnatal changes in the Rexed lamination and markers of nociceptive afferents in the superficial dorsal horn of the rat. AB - In this study, we investigated postnatal changes in Rexed's laminae and distribution of nociceptive afferents in the dorsal horn of the rat lumbar spinal cord at postnatal days 0, 5, 10, 15, 20, and 60. Transverse sections of the L4-L5 segments were processed for triple labeling with isolectin B4 (IB4)-binding as a marker of nonpeptidergic C-fibers, calcitonin gene-related peptide (CGRP) immunoreactivity to label peptidergic nociceptive afferents, and a fluorescent Nissl stain to visualize cells and lamination at different stages of postnatal development. The Nissl staining revealed that the thickness of lamina I (LI) and outer lamina II remained mostly unchanged from birth until adulthood. CGRP afferents terminated mostly in LI and the outer two-thirds of lamina II, whereas the termination area of fibers binding IB4 was centered on the middle one-third of lamina II at all ages studied. In absolute values, the overall width of the bands of intense CGRP and IB4 labeling increased with age but decreased as a percentage of the overall thickness of the dorsal horn with maturation. The overlap of CGRP termination area with that of IB4 afferents increased with age. The consequences of these findings are twofold. First, the size of the different laminae does not grow evenly across the dorsal horn. Second, CGRP and IB4 labeling cannot be considered per se to be reliable markers of lamination during development. These findings have implications for comparing data obtained in immature and mature tissues with respect to localization of structures in the dorsal horn. PMID- 18383052 TI - Distribution of neurofilament proteins in the lateral geniculate nucleus, primary visual cortex, and area MT of adult Cebus monkeys. AB - We investigated the distribution pattern of SMI-32-immunopositive cells in the lateral geniculate nucleus (LGN) and in the primary (V1) and middle temporal (MT) cortical visual areas of the adult New World monkey Cebus apella. In the LGN, the reaction for SMI-32 labeled cells in both the magnocellular (M) and parvocellular (P) layers. However, the cellular label was heavier in M layers, which also showed a more intense labeling in the neuropil. In V1, the reaction showed a lamination pattern, with the heaviest labeling occurring in layer 4B and upper layer 6 (layers that project to area MT). Area MT shows a dense band of labeled neuropil and large pyramidal neurons in layer 3, large darkly labeled but less densely packed neurons in layer 5, and a population of small, lightly labeled cells in layer 6. These results resemble those found in other New and Old World monkeys, which suggest that the preferential labeling of projection neurons associated with fast-conducting pathways to the extrastriate dorsal stream is a common characteristic of simian primates. In the superficial layers of V1 in Cebus monkeys, however, SMI-32-labeled neurons are found in both cytochrome oxidase blobs and interblob regions. In this aspect, our results in Cebus are similar to those found in the Old World monkey Macaca and different from those described for squirrel monkey, a smaller New World Monkey. In Cebus, as well as in Macaca, there is no correlation between SMI-32 distribution and the blob pattern. PMID- 18383054 TI - Hexagonal-shaped tin glycolate particles: a preliminary study of their suitability as li-ion insertion electrodes. AB - Tin glycolate particles were prepared by a simple, one-step, polyol-mediated synthesis in air in which tin oxalate precursor was added to ethylene glycol and heated at reflux. Hexagonal-shaped, micron-sized tin glycolate particles were formed when the solution had cooled. A series of tin oxides was produced by calcination of the synthesized tin glycolate at 600-800 degrees C. It was revealed that the micron-sized, hexagonal-shaped tin glycolate now consisted of nanosized tin-based particles (80-120 nm), encapsulated within a tin glycolate shell. XRD, TGA, and FT-IR measurements were conducted to account for the three dimensional growth of the tin glycolate particles. When applied as an anode material for Li-ion batteries, the synthesized tin glycolate particles showed good electrochemical reactivity in Li-ion insertion/deinsertion, retaining a specific capacity of 416 mAh g(-1) beyond 50 cycles. This performance was significantly better than those of all the other tin oxides nanoparticles (<160 mAh g(-1)) obtained after heat treatment in air. We strongly believe that the buffering of the volume expansion by the glycolate upon Li-Sn alloying is the main factor for the improved cycling of the electrode. PMID- 18383053 TI - Corpus callosum connections of subdivisions of motor and premotor cortex, and frontal eye field in a prosimian primate, Otolemur garnetti. AB - The callosal connections of motor and premotor fields in the frontal cortex of galagos were examined by placing multiple tracers into the primary motor area (M1), dorsal premotor area (PMD), ventral premotor area (PMV), supplementary motor area (SMA), and frontal eye field (FEF) following intracortical microstimulation. Retrogradely labeled neurons in the opposite hemisphere were plotted and superimposed onto brain sections stained with myelin and cytochrome oxidase for architectonic analysis. The main callosal connections of M1 and the caudal portion of PMD (PMDc) were with homotopic sites, and the major callosal connections of the rostral portion of PMD (PMDr), SMA, and FEF were with homotopic sites and adjoining cortex in the frontal lobe. In addition, M1 forelimb representation had sparse callosal connections, whereas M1 trunk and face representations, as well as the premotor areas, had dense callosal connections. The sparse interhemispheric connections of the forelimb sector of M1 suggests that the control of each forelimb is largely from the contralateral M1 in galagos, as in other primates. PMID- 18383055 TI - A heterogeneous-catalyst-based, microwave-assisted protocol for the synthesis of 2,2'-bipyridines. AB - A new method of preparing 2,2'-bipyridines with short reaction times by using microwave assistance and heterogeneous catalysts has been developed. With a Negishi-like protocol, it was found that Ni/Al(2)O(3)-SiO(2) afforded 2,2' bipyridine products in up to 86% yield in 1 h. Palladium supported on alumina also provided yields of 2,2'-bipyridines comparable to those seen for homogeneous PEPPSI (1,3-diisopropylimidazol-2-ylidene)(3 chloropyridyl)palladium(II)dichloride) and tetrakis(triphenylphosphanyl)palladium complexes. PMID- 18383056 TI - Evolution in reverse: engineering a D-xylose-specific xylose reductase. PMID- 18383057 TI - An amphiphilic bisporphyrin and its Yb(III) complex: development of a bifunctional photodynamic therapeutic and near-infrared tumor-imaging agent. PMID- 18383058 TI - Osmotic pressure and phase boundary determination of multiphase systems by analytical ultracentrifugation. AB - We show that analytical ultracentrifugation can be applied to derive full equations of state of colloids in a single sedimentation equilibrium experiment, by determination of single-phase boundaries as well as of osmotic pressure versus concentration at fixed temperatures. A continuous dependence of the osmotic pressure, over orders of magnitude between at least approximately 10(1) and 10(4) Pa, and a wide concentration range, are determined in agreement with standard theoretical considerations. Two model experimental colloidal systems are investigated: For a well-known synthetic clay system (laponite), it is shown that two regimes-counter-ion ideal gas and interacting double layers-can easily be identified in the equation of state, whereas metastable glass- or microphase separated gel states previously encountered in osmotic stress measurements of laponite are circumvented. For the case of rigid, crystallized catanionic bilayers, single phase domains can be identified. Osmotic pressure results in this case disagree with results obtained using the classical osmotic stress technique, as a result of sample adhesion to the ultracentrifuge cell windows and uncertainty due to possible micromolar ion contamination. PMID- 18383059 TI - Predicting C-H/pi interactions with nonlocal density functional theory. AB - We examine the performance of a recently developed nonlocal density functional in predicting a model noncovalent interaction, namely the weak bond between an aromatic pi system and an aliphatic C--H group. The new functional is a significant improvement over traditional density functionals, providing results which compare favorably to high-level quantum-chemistry techniques, but at considerably lower computational cost. Interaction energies in several model C- H/pi systems are in good general agreement with coupled-cluster calculations, though equilibrium distances are consistently overpredicted when using the revPBE functional for exchange. The new functional predicts changes in energy upon addition of halogen substituents correctly. PMID- 18383060 TI - Density functional investigations of electronic structure and dehydrogenation reactions of Al- and Si-substituted magnesium hydride. AB - The effect on the hydrogen storage attributes of magnesium hydride (MgH(2)) of the substitution of Mg by varying fractions of Al and Si is investigated by an ab initio plane-wave pseuodopotential method based on density functional theory. Three supercells, namely, 2 x 2 x 2, 3 x 1 x 1 and 5 x 1 x 1 are used for generating configurations with varying amounts (fractions x=0.0625, 0.1, and 0.167) of impurities. The analyses of band structure and density of states (DOS) show that, when a Mg atom is replaced by Al, the band gap vanishes as the extra electron occupies the conduction band minimum. In the case of Si-substitution, additional states are generated within the band gap of pure MgH(2)-significantly reducing the gap in the process. The reduced band gaps cause the Mg--H bond to become more susceptible to dissociation. For all the fractions, the calculated reaction energies for the stepwise removal of H(2) molecules from Al- and Si substituted MgH(2) are much lower than for H(2) removal from pure MgH(2). The reduced stability is also reflected in the comparatively smaller heats of formation (DeltaH(f)) of the substituted MgH(2) systems. Si causes greater destabilization of MgH(2) than Al for each x. For fractions x=0.167 of Al, x=0.1, 0.167 of Si (FCC) and x=0.0625, 0.1 of Si (diamond), DeltaH(f) is much less than that of MgH(2) substituted by a fraction x=0.2 of Ti (Y. Song, Z. X. Guo, R. Yang, Mat. Sc. & Eng. A 2004, 365, 73). Hence, we suggest the use of Al or Si instead of Ti as an agent for decreasing the dehydrogenation reaction and energy, consequently, the dehydrogenation temperature of MgH(2), thereby improving its potential as a hydrogen storage material. PMID- 18383061 TI - Direct probe of NO vibration in the naked ferric heme nitrosyl complex. PMID- 18383062 TI - Discovery of a drug-like G-quadruplex binding ligand by high-throughput docking. PMID- 18383063 TI - Naphthoxazepine inhibitors of HIV-1 integrase: synthesis and biological evaluation. AB - Two sets of compounds derived from the fusion of a diversely annulated naphthoxazepinedione system with 1,3-thiazole and 1,3-oxazole are described. These compounds are close analogues of previously reported thiazolothiazepine inhibitors of human immunodeficiency virus type 1 integrase (HIV-1 IN). Some of the new derivatives show potency similar to that of the reference compounds, thus gaining further insight into the structure-activity relationship of this class of IN inhibitors. PMID- 18383064 TI - Use of N-methylpiperazine for the preparation of piperazine-based unsymmetrical bis-ureas as anti-HIV agents. PMID- 18383065 TI - Risk of congenital malformations among infants exposed to antidepressants during pregnancy. PMID- 18383066 TI - Translating stem cells to the clinic: the good, the bad, and the ugly. PMID- 18383067 TI - Is varicella zoster virus really involved in the pathogenesis of multiple sclerosis? PMID- 18383068 TI - Transient epileptic amnesia: old syndrome or new? PMID- 18383069 TI - Rituximab in relapsing-remitting multiple sclerosis: a 72-week, open-label, phase I trial. AB - We evaluated the safety, tolerability, pharmacodynamics, and activity of B-cell depletion with rituximab in patients with relapsing-remitting multiple sclerosis, receiving two courses of rituximab 6 months apart, and followed for a total of 72 weeks. No serious adverse events were noted; events were limited to mild-to moderate infusion-associated events, which tended to decrease with subsequent infusions. Infections were also mild or moderate, and none led to withdrawal. Fewer new gadolinium-enhancing or T2 lesions were seen starting from week 4 and through week 72. An apparent reduction in relapses was also observed over the 72 weeks compared with the year before therapy. PMID- 18383071 TI - Are adenosine antagonists, such as istradefylline, caffeine, and chocolate, useful in the treatment of Parkinson's disease? PMID- 18383072 TI - Repairing the human brain after stroke: I. Mechanisms of spontaneous recovery. AB - Stroke remains a leading cause of adult disability. Some degree of spontaneous behavioral recovery is usually seen in the weeks after stroke onset. Variability in recovery is substantial across human patients. Some principles have emerged; for example, recovery occurs slowest in those destined to have less successful outcomes. Animal studies have extended these observations, providing insight into a broad range of underlying molecular and physiological events. Brain mapping studies in human patients have provided observations at the systems level that often parallel findings in animals. In general, the best outcomes are associated with the greatest return toward the normal state of brain functional organization. Reorganization of surviving central nervous system elements supports behavioral recovery, for example, through changes in interhemispheric lateralization, activity of association cortices linked to injured zones, and organization of cortical representational maps. A number of factors influence events supporting stroke recovery, such as demographics, behavioral experience, and perhaps genetics. Such measures gain importance when viewed as covariates in therapeutic trials of restorative agents that target stroke recovery. PMID- 18383074 TI - T2 clinical research: judgment day. PMID- 18383075 TI - A treatment for neurally mediated syncope? (Don't) hold your breath. PMID- 18383076 TI - Comparison of SHF and SSF processes from steam-exploded wheat straw for ethanol production by xylose-fermenting and robust glucose-fermenting Saccharomyces cerevisiae strains. AB - In this study, bioethanol production from steam-exploded wheat straw using different process configurations was evaluated using two Saccharomyces cerevisiae strains, F12 and Red Star. The strain F12 has been engineerically modified to allow xylose consumption as cereal straw contain considerable amounts of pentoses. Red Star is a robust hexose-fermenting strain used for industrial fuel ethanol fermentations and it was used for comparative purposes. The highest ethanol concentration, 23.7 g/L, was reached using the whole slurry (10%, w/v) and the recombinant strain (F12) in an SSF process, it showed an ethanol yield on consumed sugars of 0.43 g/g and a volumetric ethanol productivity of 0.7 g/L h for the first 3 h. Ethanol concentrations obtained in SSF processes were in all cases higher than those from SHF at the same conditions. Furthermore, using the whole slurry, final ethanol concentration was improved in all tests due to the increase of potential fermentable sugars in the fermentation broth. Inhibitory compounds present in the pretreated wheat straw caused a significantly negative effect on the fermentation rate. However, it was found that the inhibitors furfural and HMF were completely metabolized by the yeast during SSF by metabolic redox reactions. An often encountered problem during xylose fermentation is considerable xylitol production that occurs due to metabolic redox imbalance. However, in our work this redox imbalance was counteracted by the detoxification reactions and no xylitol was produced. PMID- 18383077 TI - First, do no harm: The question of liver biopsy in living liver donors. PMID- 18383078 TI - Antibody to hepatitis B surface antigen trough levels and half-lives do not differ after intravenous and intramuscular hepatitis B immunoglobulin administration after liver transplantation. AB - Hepatitis B immunoglobulin (HBIG) administration remains an essential component of standard reinfection prophylaxis after liver transplantation for hepatitis B virus-related liver disease. Previous studies have suggested that intramuscular (IM) HBIG administration compared to intravenous (IV) HBIG administration may be cost-effective and dose-saving. To compare antibody against hepatitis B surface antigen (anti-HBs) kinetics after IV HBIG administration versus IM HBIG administration, 24 patients received 2000 IU of HBIG every 6 weeks over a study period of 48 weeks in a crossover design. HBIG was started intravenously in 12 patients (group A) and intramuscularly in 12 patients (group B). After 4 doses, at week 24 HBIG administration was switched from IM to IV and vice versa. Anti HBs kinetics of 22 patients were evaluated. Mean anti-HBs levels measured 2, 4, and 6 weeks after each HBIG administration did not differ significantly (480 +/- 166, 319 +/- 126, and 221 +/- 106 IU/L after IV administration versus 457 +/- 166, 310 +/- 147, and 218 +/- 112 IU/L after IM administration). Half-lives of anti-HBs decline (IV, 25.5 +/- 6.0 days, versus IM, 24.7 +/- 6.2 days) and area under the curve values from week 2 to 6 (IV, 9.4 +/- 3.6 IU*day/mL, versus IM, 9.0 +/- 3.9 IU*day/mL) also showed no significant difference. Variation of anti HBs levels after IV HBIG administration versus IM HBIG administration was neither significantly different within patients (intraindividual variance) nor between patients (interindividual variance). However, intraindividual variance was lower than interindividual variance after IV (P < 0.05) and IM (P < 0.05) HBIG administration at every time point (2, 4, and 6 weeks). In conclusion, IV HBIG administration and IM HBIG administration are equally effective with respect to the crucial pharmacokinetic parameters. That is, IM HBIG is not dose-saving; however, it may be cost-effective if the price per unit is lower. Individualized dosing intervals should be further evaluated as a cost-effective alternative to fixed dosing schemes. PMID- 18383079 TI - The impact of postreperfusion syndrome on short-term patient and liver allograft outcome in patients undergoing orthotopic liver transplantation. AB - The greatest part of liver allograft injury occurs during reperfusion, not during the cold ischemia phase. The aim of this study, therefore, was to investigate how the severity of postreperfusion syndrome (PRS) influences short-term outcome for the patient and for the liver allograft. Over a 2-year period, 338 consecutive patients who presented for orthotopic liver transplantation (OLT) were included in this retrospective study. They were divided into 2 groups according to the severity of the PRS they experienced. The first group comprised 152 patients with mild or no PRS; the second group comprised 186 patients with significant PRS. Perioperative hemodynamic parameters, coagulation profiles, blood product requirements, incidence of infection, incidence of rejection and outcome data for both groups were collected and analyzed. There was no demographic difference between the groups except for age; group 2 had older patients than group 1 (54.94 +/- 9.07 versus 51.52 +/- 9.91, P = 0.001). Compared to group 1, group 2 patients required more red blood cell transfusions (11.31 +/- 10.90 versus 8.08 +/- 7.89 units, P = 0.002), more fresh frozen plasma transfusions (10.25 +/- 10.96 versus 7.03 +/- 7.64 units, P = 0.002), more cryoprecipitate (1.88 +/- 4.72 units versus 0.61 +/- 1.80 units, P = 0.001), and were more likely to suffer from fibrinolysis (52.7% versus 41.4%, P = 0.041). Interestingly, group 2 had a shorter average warm ischemia time than group 1 (33.19 +/- 8.55 versus 36.21 +/- 11.83 minutes, P = 0.01). Group 2 also required longer, on average, mechanical ventilation (14.95 +/- 29.79 versus 8.55 +/- 17.79 days, P = 0.015), remained in the intensive care unit longer (17.65 +/- 31.00 versus 11.49 +/- 18.67 days, P = 0.025), and had a longer hospital stay (27.29 +/- 32.35 versus 20.85 +/- 21.08 days, P = 0.029). Group 2 was more likely to require retransplantation (8.6% versus 3.3%, P = 0.044). In conclusion, the severity of PRS during OLT appears to be related to the outcome of patient and liver allograft. PMID- 18383080 TI - Audiometric changes in patients undergoing liver transplantation using distinct immunosuppressive protocols. AB - The aim of the study was to disclose a possible association of hearing defects with the use of either cyclosporin (CyA) or tacrolimus (FK-506) in patients undergoing liver transplantation (LT). Pure-tone audiometry (PTA) was performed in the same 42 patients before and after LT. Audiometric frequencies recorded ranged from 250 to 8000 Hz. Patients were divided into 2 groups according to the use of CyA (n = 18) or FK-506 (n = 24). We used Kolmogorov-Smirnov and subsequently Student t test, nonparametric Wilcoxon signed-rank test, and Mann Whitney test for statistical analysis. Significance was achieved when P 1,400) for an intragenic recombination hot spot in both genes. The 121ins2 SFTPB mutation occurred predominantly (89%) on 2 common haplotypes. In contrast, no consistent haplotypes were associated with mutated SFTPC alleles. Sporadic SFTPC mutations arose on the paternal allele in four of five families; the remaining child had evidence for somatic recombination on the mutated allele. CONCLUSIONS: In contrast to SFTPB, disease alleles at SFTPC do not share a common haplotype background. Most sporadic mutations in SFTPC occurred on the paternal allele, but somatic recombination may be an important mechanism of mutation in SFTPC. PMID- 18383114 TI - Speech rate and rhythm in Parkinson's disease. AB - Articulatory rate and pause time in a standardized reading task in Parkinson's disease (PD) patients in correlation to disease duration and severity as compared to healthy controls were analyzed. In 121 PD patients and 70 healthy controls, an acoustical analysis was performed on the first and last sentence of a standardized 170-syllabic text, using a commercial audio software. Articulatory rate and speech to pause ratios were calculated by measuring the length of each syllable and each pause both at the end of words and within polysyllabic words. No significant difference in overall articulatory rate was found between PD patients and controls. Both groups showed an accelerated speech rate in the last sentence compared to the first; however, PD patients had a higher speech acceleration than did controls. PD patients exhibited a significantly reduced percental pause duration in relation to total speech time in the first sentence and a reduced percental pause time within polysyllabic words. PD patients made significantly less but longer pauses at the end of words and less pauses within polysyllabic words. UPDRS III showed an inverse relation to number and rate of intraword pauses, and disease duration was negatively correlated with articulatory rate. The characteristics of parkinsonian speech feature was not only a stronger acceleration of articulation rate in the course of speaking but also a significant reduction of the total numbers of pauses, indicating an impaired speech rhythm and timing organization. PMID- 18383115 TI - Pulse rate and pulse rate variability decrease after adenotonsillectomy for obstructive sleep apnea. AB - BACKGROUND: Data suggest that obstructive sleep apnea syndrome (OSA) results in sympathetic stimulation, brady/tachycardia and cardiac stress. Heart rate variability, but not baseline heart rate, is known to be elevated in pediatric OSA. Our patients with moderate to severe OSA (McGill Oximetry Scores of 3 or 4) have been re-evaluated with pulse oximetry after adenotonsillectomy (T&A). We hypothesized that pulse rate (PR) and pulse rate variability (PRV) would decrease after treatment of OSA with T&A. METHODS: This retrospective before-after study comprised pre- and post-operative oximetries and parental questionnaires of children 1-18 years old with moderate to severe OSA from September 2004 to August 2005, inclusive. We excluded patients with significant comorbidities. RESULTS: In 25 subjects, age at surgery was 4.3 +/- 3.6 years (mean +/- SD). OSA symptoms decreased or resolved, saturation metrics improved, and parental concern about breathing during sleep decreased following T&A. PR decreased in 21 of 25 patients after T&A (mean PR from 99.7 +/- 11.2 to 90.1 +/- 10.7 bpm, P < 0.001; maximum PR from 150.6 +/- 14.5 to 137.4 +/- 15.6 bpm, P < 0.001). PRV, as measured by the standard deviation of the PR, decreased in 23 of 25 patients after T&A (from 10.3 +/- 2.1 to 8.2 +/- 1.6 bpm, [P < 0.001]). Pulse accelerations greater than 6, 7, 8 bpm also decreased post-operatively. CONCLUSIONS: Nocturnal pulse oximetry complements clinical history to document improvement and/or resolution of moderate to severe OSA in children. Resolution of tachycardia and diminished PRV after T&A illustrate the stress that recurrent airway obstruction during sleep places on the cardiovascular system. Further work will be required to determine if PR and PRV as measured by pulse oximetry would be useful in the diagnosis and follow-up of OSA in children. PMID- 18383116 TI - The outcome of infants younger than 6 months requiring ventilation for pneumonia caused by Mycobacterium tuberculosis. AB - INTRODUCTION: The outcome of young infants (<6 months) being ventilated for respiratory failure caused by Mycobacterium tuberculosis (MTB) has not been recorded. PATIENTS AND METHODS: A descriptive study of children <6 months admitted to the PICU from 1 February 1999 to 31 December 2005 with MTB causing respiratory failure. RESULTS: Seventeen infants were ventilated for respiratory failure caused by MTB: ten had ventilatory respiratory failure and seven had hypoxic failure. An index case was found in 47%. All chest radiographs (CXRs) were highly suggestive of tuberculosis. MTB was cultured in 15 cases. In the other two cases MTB was confirmed by histopathology. The median duration of ventilation was 6 days (range: 1-35 days) with a median PaO2/FiO2 of 85 and ventilatory index of 58. Transthoracic glandular enucleation was required to facilitate extubation in six babies. All the infants survived. At 6-month follow up 35% had a normal CXR and all were asymptomatic. One child had CXR changes suggestive of bronchiectasis but was asymptomatic. CONCLUSION: The outcome of infants <6 months ventilated for respiratory failure caused by MTB is very good if TB is recognized timeously and appropriate management started. The diagnosis of TB in these infants can be made with a high index of suspicion and careful evaluation of the CXR. PMID- 18383117 TI - Novel approach to the eradication of Pseudomonas aeruginosa in an infant with CF after outpatient treatment failure. AB - Intravenous continuous infusion of betalactam (CIBL) antibiotic and high dose extended interval (HDEI) aminoglycoside therapy theoretically maximize bacterial killing in treatment of Pseudomonas aeruginosa (PsA) in pulmonary exacerbations of cystic fibrosis (CF). We present the case of a 3-month-old female infant with CF who failed outpatient eradication of PsA with subsequent eradication using intravenous CIBL antibiotic and HDEI aminoglycoside therapy. This antibiotic combination should be considered in order to optimize pharmacodynamics for PsA eradication in CF patients before development of chronic colonization. PMID- 18383118 TI - Regression of symptoms after selective iron chelation therapy in a case of neurodegeneration with brain iron accumulation. AB - We report the results of iron chelating treatment with deferiprone in a 61-year old woman with signs and symptoms of neurodegeneration with brain iron accumulation (NBIA). After 6 months of therapy the patient's gait had improved and a reduction in the incidence of choreic dyskinesias was observed. Her gait returned to normal after an additional 2 months of therapy, at which time there was a further reduction in involuntary movements and a partial resolution of the blepharospasm. PMID- 18383121 TI - Rotating versus perfusion bioreactor for the culture of engineered vascular constructs based on hyaluronic acid. AB - It is generally accepted that dynamic culture conditions are required for vascular tissue engineering. We compared the effects of two dynamic culture systems, a perfusion and a rotating bioreactor, using tubular constructs based on hyaluronic acid seeded with porcine aortic smooth muscle cells (SMC), that we recently showed to be adequate for the generation of vascular tissue. In perfused constructs mechanical stimulation importantly affected cell morphology, increased the incidence of cell proliferation and reduced apoptosis. However, extracellular matrix deposition, cytoskeletal organization and mechanical properties were poor. In rotated constructs cell proliferation was also higher and apoptosis lower than in static controls. Rotated constructs showed the highest ultimate stress and the lowest elastic modulus. Our data indicate that the rotating bioreactor is more efficient than the perfusion bioreactor and we then suggest that this method can be considered a valid alternative to complex bioreactor systems described in the literature. PMID- 18383122 TI - Nanostructured microspheres produced by supercritical fluid extraction of emulsions. AB - The system poly(lactic-co-glycolic) acid/ piroxicam (PLGA/PX) was selected, as a model system, to evaluate the effectiveness of supercritical carbon dioxide (SC CO(2)) extraction of the oily phase (ethyl acetate) from oil-in-water emulsions used in the production of polymer/drug microspheres for sustained drug release applications. The influence of process parameters like operating pressure and temperature, flow rate and contacting time between the emulsion and SC-CO(2) was studied with respect to the microsphere size, distribution and solvent residue. Different polymer concentrations in the oily phase were also tested in emulsions formulation to monitor their effects on droplets and microspheres size distribution at fixed mixing conditions. Spherical PLGA microspheres loaded with PX (10% w/w) with mean sizes ranging between 1 and 3 microm and very narrow size distributions were obtained due to the short supercritical processing time (30 min) that prevents the aggregation phenomena typically occurring during conventional solvent evaporation process. A solvent residue smaller than 40 ppm was also obtained at optimized operating conditions. DSC and SEM-EDX analyses confirmed that the produced microparticles are formed by a solid solution of PLGA and PX and that the drug is entrapped in an amorphous state into the polymeric matrix with an encapsulation efficiency in the range of 90-95%. Drug release rate studies showed very uniform drug concentration profiles, without any burst effect, confirming a good dispersion of the drug into the polymer particles. PMID- 18383124 TI - Multiplex bacterial growth monitoring in 24-well microplates using a dual optical sensor for dissolved oxygen and pH. AB - Non-invasive, simultaneous optical monitoring of oxygen and pH during bacterial cultivation in 24-well microplates is presented using an integrated dual sensor for dissolved oxygen and pH values. The dual sensor is based on oxygen-sensitive organosilica microparticles and pH-sensitive microbeads from a polymethacrylate derivative embedded into a polyurethane hydrogel. The readout is based on a phase domain fluorescence lifetime-based method referred to as modified frequency domain dual lifetime referencing using a commercially available detector system for 24-well microplates. The sensor was used for monitoring the growth of Pseudomonas putida bacterial cultures. The method is suitable for parallelized, miniaturized bioprocessing, and cell-based high-throughput screening applications. PMID- 18383125 TI - Methane production in an UASB reactor operated under periodic mesophilic thermophilic conditions. AB - Methane production was studied in a laboratory-scale 10 L anaerobic upflow sludge bed (UASB) reactor with periodic variations of the reactor temperature. On a daily basis the temperature was varied between 35 and 45 degrees C or 35 and 55 degrees C with a heating period of 6 h. Each temperature increase was accompanied by an increase in methane production and a decrease in the concentration of soluble organic matter in the effluent. In comparison to a reactor operated at 35 degrees C, a net increase in methane production of up to 22% was observed. Batch activity tests demonstrated a tolerance of mesophilic methanogenic populations to short-term, 2-6 h, temperature increases, although activity of acetoclastic methanogens decreased after 6 h exposure to a temperature of 55 degrees C. 16S sequencing of DGGE bands revealed proliferation of temperature-tolerant Methanospirillum hungatii sp. in the reactor. PMID- 18383123 TI - Review on genetic variants and maternal smoking in the etiology of oral clefts and other birth defects. AB - A spectrum of adverse pregnancy outcomes, including preterm birth, low birth weight, and birth defects has been linked with maternal smoking during pregnancy. This article includes a review of studies investigating interactions between genetic variants and maternal smoking in contributing to birth defects using oral clefting as a model birth defect. The primary gene-smoking studies for other major birth defects are also summarized. Gene-environment interaction studies for birth defects are still at an early stage with several mixed results, but evolving research findings have begun to document clinically and developmentally important interactions. As samples and data become increasingly available, more effort is needed in designing innovative analytical methods to study gene environment interactions. PMID- 18383126 TI - The online assignment of the absolute configuration of natural products: HPLC-CD in combination with quantum chemical CD calculations. AB - The application of modern online methods, e.g., HPLC-MS/MS and HPLC-NMR, allows the elucidation of constitutions and relative configurations of new natural products directly from crude extracts. To additionally establish the full absolute configurations of such secondary metabolites without the necessity of first isolating the compounds, we have introduced HPLC-CD coupling (CD = circular dichroism) into natural product analysis, taking advantage of the different chiroptical properties of stereoisomers, in particular of enantiomers. In combination with quantum chemical CD calculations this method allows the stereochemical characterization of (even structurally unprecedented) chiral molecules, thus avoiding the--often risky--merely empirical assignment by comparison with the CD spectra of related compounds with known absolute stereostructures, or by other methods such as, e.g., the exciton chirality approach. This review presents the experimental requirements for the hyphenation and the theoretical background of the calculation of UV and CD spectra, which is then exemplified by some recent HPLC-CD applications to the elucidation of absolute configurations of most diverse compounds of mainly natural origin. PMID- 18383127 TI - Utilization of cellulosic waste from tequila bagasse and production of polyhydroxyalkanoate (PHA) bioplastics by Saccharophagus degradans. AB - Utilization of wastes from agriculture is becoming increasingly important due to concerns of environmental impact. The goals of this work were to evaluate the ability of an unusual organism, Saccharophagus degradans (ATCC 43961), to degrade the major components of plant cell walls and to evaluate the ability of S. degradans to produce polyhydroxyalkanoates (PHAs, also known as bioplastics). S. degradans can readily attach to cellulosic fibers, degrade the cellulose, and utilize this as the primary carbon source. The growth of S. degradans was assessed in minimal media (MM) containing glucose, cellobiose, avicel, and bagasse with all able to support growth. Cells were able to attach to avicel and bagasse fibers; however, growth on these insoluble fibers was much slower and led to a lower maximal biomass production than observed with simple sugars. Lignin in MM alone did not support growth, but did support growth upon addition of glucose, although with an increased adaptation phase. When culture conditions were switched to a nitrogen depleted status, PHA production commences and extends for at least 48 h. At early stationary phase, stained inclusion bodies were visible and two chronologically increasing infrared light absorbance peaks at 1,725 and 1,741 cm(-1) confirmed the presence of PHAs. This work demonstrates for what we believe to be the first time, that a single organism can degrade insoluble cellulose and under similar conditions can produce and accumulate PHA. Additional work is necessary to more fully characterize these capabilities and to optimize the PHA production and purification. PMID- 18383128 TI - In vitro assessment of reproductive toxicity of tobacco smoke and its constituents. AB - Epidemiological studies have repeatedly shown that reproductive processes in pregnant women are adversely affected by exposure to cigarette smoke. The potential reproductive targets of smoke during pregnancy include the ovaries, oviducts, uterus, placenta, umbilical cord, and embryo/fetus. In vitro methods for studying the effects of smoke and its individual components have been developed and applied to each of these reproductive targets. In vitro assays have been useful in determining the biological processes that are affected in the reproductive organs and in identifying the cellular and molecular targets of smoke in each organ. In vitro methods have also been used to study the mechanism of action of smoke constituents, such as nicotine, on specific processes in reproductive organs and to screen smoke solutions to identify the molecules that affect reproduction. In general, data collected in vitro have confirmed, extended, and helped explain what has been learned from epidemiological studies. This review summarizes some of the in vitro assays that have been used to study cigarette smoke's effect on the nonpregnant and pregnant female reproductive tract and spotlights examples of their applications. PMID- 18383129 TI - Reducing harm from tobacco smoke exposure during pregnancy. AB - In addition to the health risks that maternal tobacco smoke exposure in pregnancy poses to women, this is a cause of substantial fetal morbidity and mortality. In pregnancy, maternal tobacco smoke exposure can arise because women either smoke or are passively exposed to environmental tobacco smoke as a consequence of other's smoking. This article discusses the scope for clinicians to help reduce both types of tobacco smoke exposure in pregnancy, with a specific focus on available and effective interventions for smoking cessation by pregnant women. Behavioral support with smoking cessation is the only intervention that has been proven to encourage smoking cessation in pregnancy and reduces smoking rates in late pregnancy by 6 to 7%. There are physiological reasons to suspect that nicotine replacement therapy (NRT) will be less or (in)effective for smoking cessation in pregnancy when compared with its use by nonpregnant smokers. However, there are also strong theoretical reasons to suspect that NRT is likely to be safer than continued smoking in pregnancy. Consequently, this article reviews evidence for the safety and effectiveness of NRT when used for smoking cessation in pregnancy and recommendations concerning the use of NRT in pregnancy are presented. PMID- 18383130 TI - Nicotine and brain development. AB - Preclinical studies, using primarily rodent models, have shown acetylcholine to have a critical role in brain maturation via activation of nicotinic acetylcholine receptors (nAChRs), a structurally diverse family of ligand-gated ion channels. nAChRs are widely expressed in fetal central nervous system, with transient upregulation in numerous brain regions during critical developmental periods. Activation of nAChRs can have varied developmental influences that are dependent on the pharmacologic properties and localization of the receptor. These include regulation of transmitter release, gene expression, neurite outgrowth, cell survival, and synapse formation and maturation. Aberrant exposure of fetal and neonatal brain to nicotine, through maternal smoking or nicotine replacement therapy (NRT), has been shown to have detrimental effects on cholinergic modulation of brain development. These include alterations in sexual differentiation of the brain, and in cell survival and synaptogenesis. Long-term alterations in the functional status and pharmacologic properties of nAChRs may also occur, which result in modifications of specific neural circuitry such as the brainstem cardiorespiratory network and sensory thalamocortical gating. Such alterations in brain structure and function may contribute to clinically characterized deficits that result from maternal smoking, such as sudden infant death syndrome and auditory-cognitive dysfunction. Although not the only constituent of tobacco smoke, there is now abundant evidence that nicotine is a neural teratogen. Thus, alternatives to NRT should be sought as tobacco cessation treatments in pregnant women. PMID- 18383131 TI - Nicotine and lung development. AB - Nicotine is found in tobacco smoke. It is a habit forming substance and is prescribed by health professionals to assist smokers to quit smoking. It is rapidly absorbed from the lungs of smokers. It crosses the placenta and accumulates in the developing fetus. Nicotine induces formation of oxygen radicals and at the same time also reduces the antioxidant capacity of the lungs. Nicotine and the oxidants cause point mutations in the DNA molecule, thereby changing the program that controls lung growth and maintenance of lung structure. The data available indicate that maternal nicotine exposure induces a persistent inhibition of glycolysis and a drastically increased cAMP level. These metabolic changes are thought to contribute to the faster aging of the lungs of the offspring of mothers that are exposed to nicotine via the placenta and mother's milk. The lungs of these animals are more susceptible to damage as shown by the gradual deterioration of the lung parenchyma. The rapid metabolic and structural aging of the lungs of the animals that were exposed to nicotine via the placenta and mother's milk, and thus during phases of lung development characterized by rapid cell division, is likely due to "programming" induced by nicotine. It is, therefore, not advisable to use nicotine during gestation and lactation. PMID- 18383132 TI - Detrimental effects of tobacco smoke exposure during development on postnatal lung function and asthma. AB - Exposure to environmental tobacco smoke (ETS) during fetal development and early postnatal life is perhaps the most ubiquitous and hazardous of children's environmental exposures. The developing lung is highly susceptible to ETS. A large body of literature links both prenatal maternal smoking and children's ETS exposure to decreased lung growth. This review summarizes the state of the knowledge, including both human epidemiology and laboratory animal experiments, linking ETS, lung development, and respiratory outcomes. Important issues discussed include lung development and lung function and asthma in relation to ETS exposure during critical windows of growth. Prenatal exposure to ETS is associated with impaired lung function and increased risk of developing asthma, whereas postnatal exposure mainly acts to trigger respiratory symptoms and asthma attacks, but it also plays an important role in the occurrence of asthma in children. This review provides evidence that avoidance of ETS exposure both before and after birth is beneficial to long-term respiratory health, because airway function in later life is believed to be largely determined by lung development occurring in utero and in early infancy. PMID- 18383133 TI - Tobacco and pregnancy: overview of exposures and effects. AB - This opening article will review the epidemiology of the effects of cigarette smoking and other forms of tobacco exposure on human development. Sources of exposure described include cigarettes and other forms of smoked tobacco, secondhand (environmental) tobacco smoke, several forms of smokeless tobacco, and nicotine from nicotine replacement therapy. Exposure is immense and worldwide, most of it due to smoking, but in some parts of the world and in some populations, smoking is exceeded by smokeless tobacco use. Nicotine and carbon monoxide exposure are of large concern, but cigarette smoke contains over 4000 chemical constituents and additives including known carcinogens, toxic heavy metals, and many chemicals untested for developmental toxicity. The impact of tobacco on human development will be reviewed. Fertility, conception, survival of the conceptus, most phases and aspects of development studied to date, as well as postnatal survival and health are adversely impacted by maternal tobacco use or exposure. Effects in surviving offspring are probably life-long, and are still being elucidated. It is hoped that this review and those to follow in this issue will serve to keep a focus on the critical and continuing problem of tobacco use impacting human development. PMID- 18383134 TI - Concerning about the accuracy of data reported. PMID- 18383135 TI - Role of the tumor necrosis factor-alpha, cyclooxygenase-2, prostaglandin E2, and effect of low-intensity pulsed ultrasound in an in vitro herniated disc resorption model. AB - Spontaneous herniated disc resorption occurs via inflammatory reactions involving abundant neovascularization and macrophage phagocytotic activity. Nonthermal low intensity pulsed ultrasound (LIPUS) treatment might be effective in shortening the duration of disc resorption. We developed a rat in vitro resorption model in which a coccygeal intervertebral disc and peritoneal macrophages were cocultured. Secretion of tumor necrosis factor-alpha (TNF-alpha) from macrophages was promoted by LIPUS, and the process of disc degeneration was thus accelerated. In this study, we further examined the effects of LIPUS using this in vitro model focusing on whether LIPUS affects cyclooxygenase-2 (COX-2) signaling pathways. We found that the levels of COX-2 and prostaglandin E2 (PGE2) secreted from macrophages were increased by LIPUS. However, these phenomena were not caused by LIPUS directly, as the levels of these substances were reduced by neutralizing TNF-alpha activity. Moreover, the wet weights of the disc samples were not changed by addition of PGE2, but were reduced by recombinant TNF-alpha. Our results suggest that the effects of LIPUS in enhancing the process of herniated disc resorption are caused mainly by TNF-alpha. PMID- 18383138 TI - On the calculation of the elastic modulus of a biofilm streamer. AB - Biofilm mechanical properties are essential in quantifying the rate of microbial detachment, a key process in determining the function and structure of biofilm systems. Although properties such as biofilm elastic moduli, yield stress and cohesive strength have been studied before, a wide range of values for the biofilm Young's modulus that differ by several orders of magnitude are reported in the literature. In this article, we use experimental data reported in Stoodley et al. [Stoodley et al., Biotechnol Bioeng (1999): 65(1):83-92] and present a methodology for the calculation of Young's modulus, which partially explains the large difference between the values reported in the literature. PMID- 18383139 TI - Ionic liquids as alternative co-solvents for laccase: study of enzyme activity and stability. AB - The activity and stability of commercial laccase (DeniLite base) in three different water soluble ionic liquids (ILs) (1-ethyl-3-methylimidazolium 2-(2 methoxyethoxy) ethylsulfate, [emim][[MDEGSO4], 1-ethyl-3-methylimidazolium ethylsulfate, [emim][EtSO4], and 1-ethyl-3-methylimidazolium methanesulfonate, [emim][MeSO3]) have been studied and compared to that in two organic solvents (acetonitrile and dimethyl sulfoxide). Initial enzyme activities were similar among the ILs if the same conditions were used. A high reduction on initial enzyme activity was found with acidic pH (5.0). The effect of pH and solvent concentration on enzyme stability were investigated in more detail for 1 week. The enzyme maintained a high stability at pH 9.0 for all ILs tested. [emim][MDEGSO4] was the most promising IL for laccase with an activity loss of about 10% after 7 days of incubation. The kinetic studies in the presence of ABTS as substrate allowed to calculate the Michaelis- Menten parameters. Good agreement was found between experimental data and calculated values using the Michaelis-Menten mechanism, with a total average relative deviation of 2.1%. PMID- 18383140 TI - Determination of the cytosolic free NAD/NADH ratio in Saccharomyces cerevisiae under steady-state and highly dynamic conditions. AB - The coenzyme NAD plays a major role in metabolism as a key redox carrier and signaling molecule but current measurement techniques cannot distinguish between different compartment pools, between free and protein-bound forms and/or between NAD(H) and NADP(H). Local free NAD/NADH ratios can be determined from product/substrate ratios of suitable near-equilibrium redox reactions but the application of this principle is often precluded by uncertainties regarding enzyme activity, localization and coenzyme specificity of dehydrogenases. In Saccharomyces cerevisiae, we circumvented these issues by expressing a bacterial mannitol-1-phosphate 5-dehydrogenase and determining the cytosolic free NAD/NADH ratio from the measured [fructose-6-phosphate]/[mannitol-1-phosphate] ratio. Under aerobic glucose-limited conditions we estimated a cytosolic free NAD/NADH ratio between 101(+/-14) and 320(+/-45), assuming the cytosolic pH is between 7.0 and 6.5, respectively. These values are more than 10-fold higher than the measured whole-cell total NAD/NADH ratio of 7.5(+/-2.5). Using a thermodynamic analysis of central glycolysis we demonstrate that the former are thermodynamically feasible, while the latter is not. Furthermore, we applied this novel system to study the short-term metabolic responses to perturbations. We found that the cytosolic free NAD-NADH couple became more reduced rapidly (timescale of seconds) upon a pulse of glucose (electron-donor) and that this could be reversed by the addition of acetaldehyde (electron-acceptor). In addition, these dynamics occurred without significant changes in whole-cell total NAD and NADH. This approach provides a new experimental tool for quantitative physiology and opens new possibilities in the study of energy and redox metabolism in S. cerevisiae. The same strategy should also be applicable to other microorganisms. PMID- 18383141 TI - Sensitivity of knee replacement contact calculations to kinematic measurement errors. AB - The ability to measure in vivo knee kinematics accurately makes it tempting to calculate in vivo contact forces, pressures, and areas directly from kinematic data. However, the sensitivity of contact calculations to kinematic measurement errors has not been adequately investigated. To address this issue, we developed a series of sensitivity analyses derived from a validated in vivo computational simulation of gait. The simulation used an elastic foundation contact model to reproduce in vivo contact force, center of pressure, and fluoroscopic motion data collected from an instrumented knee replacement. Treating each degree of freedom (DOF) in the simulation as motion controlled, we first quantified how errors in measured relative pose of the implant components affected contact calculations. Pose variations of +/-0.1 mm or degree over the entire gait cycle changed maximum contact force, pressure, and area by 204, 100, and 117%, respectively. Larger variations of +/-0.5 mm or degree changed these same quantities by 1157, 108, and 578%, respectively. In both cases, the largest sensitivities were to errors in superior-inferior translation and varus-valgus rotation, with loss of contact occurring on one or both sides. We then quantified how switching the sensitive DOFs from motion to load control affected the sensitivity results. Pose variations of +/-0.5 mm or degree in the remaining DOFs changed maximum contact quantities by at most 3%. These results suggest that accuracy on the order of microns and milliradians is needed to estimate contact forces, pressures, and areas directly from in vivo kinematic measurements, and that use of load rather than motion control for the sensitive DOFs may improve the accuracy of in vivo contact calculations. PMID- 18383142 TI - Biohydrogen production from glucose in upflow biofilm reactors with plastic carriers under extreme thermophilic conditions (70 degrees C). AB - Biohydrogen could efficiently be produced in glucose-fed biofilm reactors filled with plastic carriers and operated at 70 degrees C. Batch experiments were, in addition, conducted to enrich and cultivate glucose-fed extreme-thermophilic hydrogen producing microorganisms from a biohydrogen CSTR reactor fed with household solid waste. Kinetic analysis of the biohydrogen enrichment cultures show that substrate (glucose) likely inhibited hydrogen production when its concentration was higher than 1 g/L. Different start up strategies were applied for biohydrogen production in biofilm reactors operated at 70 degrees C, and fed with synthetic medium with glucose as the only carbon and energy source. A biofilm reactor, started up with plastic carriers, that were previously inoculated with the enrichment cultures, resulted in higher hydrogen yield (2.21 mol H(2)/mol glucose consumed) but required longer start up time (1 month), while a biofilm reactor directly inoculated with the enrichment cultures reached stable state much faster (8 days) but with very low hydrogen yield (0.69 mol H(2)/mol glucose consumed). These results indicate that hydraulic pressure is necessary for successful immobilization of bacteria on carriers, while there is the risk of washing out specific high yielding bacteria. PMID- 18383143 TI - A photobioreactor system for precision cultivation of photoautotrophic microorganisms and for high-content analysis of suspension dynamics. AB - Small-scale photobioreactors for cultivation of photoautotrophic microbes are required for precise characterization of the growth parameters of wild-type and engineered strains of these organisms, for their screening, and for optimization of culture conditions. Here, we describe the design and use of a flat-cuvette photobioreactor that allows accurate control of culture irradiance, temperature, pH, and gas composition combined with real-time monitoring by a built-in fluorometer and densitometer. The high-power LED light source generates precise irradiance levels that are programmed by user-designed protocols. The irradiance, temperature, and gas composition may be static or dynamically modulated, while optical density and pH may be stabilized in turbidostat and pH-stat modes, respectively. We demonstrate that the instrument is able to detect minute variations of growth caused, for example, by sudden dilution or by circadian rhythms. The sensitivity of the instrument is sufficient to monitor suspension optical density as low as 10(-2). This newly designed photobioreactor can significantly contribute to the study and use of photoautotrophic microbes in systems biology and biotechnology. PMID- 18383144 TI - Effect of wrist posture on carpal tunnel pressure while typing. AB - Long weekly hours of keyboard use may lead to or aggravate carpal tunnel syndrome. The effects of typing on fluid pressure in the carpal tunnel, a possible mediator of carpal tunnel syndrome, are unknown. Twenty healthy subjects participated in a laboratory study to investigate the effects of typing at different wrist postures on carpal tunnel pressure of the right hand. Changes in wrist flexion/extension angle (p = 0.01) and radial/ulnar deviation angle (p = 0.03) independently altered carpal tunnel pressure; wrist deviations in extension or radial deviation were associated with an increase in pressure. The activity of typing independently elevated carpal tunnel pressure (p = 0.001) relative to the static hand held in the same posture. This information can guide the design and use of keyboards and workstations in order to minimize carpal tunnel pressure while typing. The findings may also be useful to clinicians and ergonomists in the management of patients with carpal tunnel syndrome who use a keyboard. PMID- 18383145 TI - ASARM-truncated MEPE and AC-100 enhance osteogenesis by promoting osteoprogenitor adhesion. AB - Matrix extracellular phosphoglycoprotein (MEPE) is a member of the SIBLING (Small Integrin-Binding Ligand, N-linked Glycoprotein) family of secreted glycophosphoproteins. Several previous studies have demonstrated that MEPE and its peptide motif, AC-100, may regulate bone mass and influence osteoblast activity, suggesting its potential for inclusion in novel therapeutic strategies aimed at increasing osteogenesis. Our study uses in vitro approaches to assess how adhesion of nonadherent cells is influenced by MEPE and whether response to MEPE is dependent on the maturity of osteoblastic cells. Truncated MEPE (ASARM removed) or AC-100 enhanced the adhesion, spreading, and focal complex formation of unadhered osteoblastic cells leading to increased differentiation and bone formation after 28 days of culture. Furthermore, addition of truncated MEPE or AC 100 to mature osteoblasts had no significant effect on bone formation. Our data supports an action for truncated MEPE and AC-100 in altering the physiology of immature poorly adherent cells which subsequently influences the way in which these cells interact with a substrate to facilitate their survival and/or commitment to the osteoblast lineage. PMID- 18383146 TI - Management of patients with refractory migraine and PFO: Is MIST I relevant? AB - The results of the randomized clinical trial entitled: Migraine Intervention with Starflex Technology (MIST), produced surprising and disappointing results on the effect of PFO closure to decrease migraine headaches. There have been allegations of misrepresentation of the effectiveness of this device. These issues have significant implications in how randomized clinical trials are performed that will impact current and future planned trials of PFO closure to treat migraine headaches. PMID- 18383147 TI - Avascular necrosis of femoral head in childhood acute myeloid leukemia: complication of chemotherapy without steroids. PMID- 18383148 TI - Percutaneous revascularization of subclavian artery chronic occlusion with dual cerebral artery protection. AB - Percutaneous intervention for symptomatic subclavian artery chronic occlusion is an occasionally performed, minimally invasive alternative to surgical bypass. Potential complications include stroke, perforation, and recanalization failure. We discuss a case of successful percutaneous revascularization of symptomatic subclavian artery chronic occlusion using coronary devices and dual cerebral embolic protection. PMID- 18383149 TI - Day hospital versus inpatient management: An economic initiative of a pediatric center, examplified on uncomplicated vaso-occlusive crises of children with sickle cell disease. PMID- 18383150 TI - Models of tibial fracture healing in normal and Nf1-deficient mice. AB - Delayed union and nonunion are common complications associated with tibial fractures, particularly in the distal tibia. Existing mouse tibial fracture models are typically closed and middiaphyseal, and thus poorly recapitulate the prevailing conditions following surgery on a human open distal tibial fracture. This report describes our development of two open tibial fracture models in the mouse, where the bone is broken either in the tibial midshaft (mid-diaphysis) or in the distal tibia. Fractures in the distal tibial model showed delayed repair compared to fractures in the tibial midshaft. These tibial fracture models were applied to both wild-type and Nf1-deficient (Nf1+/-) mice. Bone repair has been reported to be exceptionally problematic in human NF1 patients, and these patients can also spontaneously develop tibial nonunions (known as congenital pseudarthrosis of the tibia), which are recalcitrant to even vigorous intervention. pQCT analysis confirmed no fundamental differences in cortical or cancellous bone in Nf1-deficient mouse tibiae compared to wild-type mice. Although no difference in bone healing was seen in the tibial midshaft fracture model, the healing of distal tibial fractures was found to be impaired in Nf1+/- mice. The histological features associated with nonunited Nf1+/- fractures were variable, but included delayed cartilage removal, disproportionate fibrous invasion, insufficient new bone anabolism, and excessive catabolism. These findings imply that the pathology of tibial pseudarthrosis in human NF1 is complex and likely to be multifactorial. PMID- 18383151 TI - Balloon angioplasty with stenting to correct a functionally interrupted aorta: A case report with three-year follow-up. AB - A 16-year-old male presenting with upper extremity hypertension was found to have a severe form of discrete coarctation with complete luminal obliteration, causing a functional interruption of the thoracic aorta. Fluoroscopically guided perforation of the obstruction and creation of a neo-aortic lumen was performed. This was followed by balloon angioplasty and stent placement, successfully relieving the coarctation. The procedural method, acute and late follow-up results, and a discussion of the potential risks and benefits are presented. PMID- 18383152 TI - Difficult stent delivery: use of an aspiration catheter as a "sheath". AB - This is a report of a case in which stent delivery to the lesion location proved difficult because of severely calcified and tortuous proximal vasculature. Efforts to deliver a stent using a buddy wire, anchoring technique and five-in seven guiding system were unsuccessful, whereas an aspiration catheter enabled stent delivery to the target lesion. This technique offers a potential alternative for cases in which some stent delivery methods are problematic. In addition, this technique may also prevent potential injury to drug-eluting polymer during stent delivery. PMID- 18383153 TI - Multiple intracranial tumors in Philadelphia chromosome positive acute lymphoblastic leukemia: successful treatment following aggressive supportive care, early cranial radiation, high dose chemotherapy and imatinib. AB - We report the first case of multiple intracranial tumors ("chloromas") at diagnosis of Philadelphia chromosome positive acute lymphoblastic leukemia. The patient presented comatose with signs of cerebral herniation. Initial management of raised intracranial pressure and hyperleukocytosis followed by emergent whole brain radiation therapy reversed the life-threatening neurological signs. High dose chemotherapy combined with daily imatinib mesylate induced a rapid and sustained bone marrow remission. Ongoing rehabilitation resulted in a near complete neurological recovery within 6 months of diagnosis. This outcome justifies aggressive early management of increased intracranial pressure and hyperleukocytosis in future similar presentations. PMID- 18383154 TI - Color-coded duplex ultrasound for diagnosis of renal artery stenosis and as follow-up examination after revascularization. AB - Currently, angiography is still considered to be the gold standard for the diagnosis of a renal artery stenosis (RAS). However, angiography is invasive and carries the potential risk of haematoma, pseudoaneurysm, contrast agent induced nephropathy, and athermanous embolization. Color-coded duplex ultrasound is a noninvasive frequently repeatable bed-side examination and is currently the only diagnostic method to reliably differentiate between a hemodynamically relevant or irrelevant stenosis using the side-to-side difference of the intrarenal resistance index (RI). There is a highly specific correlation between a side difference of the RI of >0.05 and an at least 70% angiographic diameter stenosis. All other duplex parameters like a peak systolic flow velocity >200 cm/sec or a renal aortic flow velocity ratio >3.5 are correlated to a 50 or 60% angiographic diameter stenosis and offer therefore indeed a high sensitivity in terms of detecting a RAS; however, the specificity detecting a hemodynamically relevant RAS is low. Provided that duplex ultrasound is performed by an experienced physician with an adequate machine it should be the preferred imaging method. The present article gives an overview about the literature related to duplex based diagnosis of RAS and as a follow-up diagnostic procedure following RAS revascularization. PMID- 18383155 TI - The chemical ecology of crucifers and their fungal pathogens: boosting plant defenses and inhibiting pathogen invasion. AB - Fungal plant diseases can cause very substantial yield losses in crucifer crops such as rapeseed and canola, or vegetables such as cabbage and broccoli. To devise sustainable methods to prevent and deter crucifer pathogens, the chemical interaction between crucifers and their fungi is under intense investigation. Crucifers produce complex blends of secondary metabolites with diverse ecological roles that include protection against microbial pathogens and other pests. The secondary metabolites involved in crucifer defense, namely phytoalexins and phytoanticipins, and their metabolism by fungal pathogens indicate that some fungi produce different enzymes to detoxify these metabolites and that some fungal detoxifying enzymes are rather specific. Chemical synthesis and screening of phytoalexin analogue libraries using cultures of fungal pathogens, as well as protein extracts, have shown that such detoxification reactions can be inhibited and that some inhibitors are strongly antifungal. Overall results of current work show the feasibility of using selective inhibitors of fungal detoxifying enzymes, i.e., paldoxins, to protect plants by boosting their chemical defenses. PMID- 18383156 TI - Maintenance versus growth: investigating the costs of immune activation among children in lowland Bolivia. AB - Immune function is a central component of maintenance effort, and it provides critical protection against the potentially life threatening effects of pathogens. However, immune defenses are energetically expensive, and the resources they consume are not available to support other activities related to growth and/or reproduction. In our study we use a life history theory framework to investigate tradeoffs between maintenance effort and growth among children in a remote area of Amazonian Bolivia. Baseline concentrations of C-reactive protein (CRP) were measured in 309 2- to 10-year olds as an indicator of immune activation, and height was measured at baseline and three months later. Elevated CRP at baseline predicts smaller gains in height over the subsequent three months, with the costs to growth particularly high for 2- to 4-year olds and for those with low energy reserves (in the form of body fat) at the time of immunostimulation. These results provide evidence for a significant tradeoff between investment in immunity and growth in humans, and highlight an important physiological mechanism through which maintenance effort may have lasting effects on child growth and development. PMID- 18383157 TI - Cross-sectional analysis of long bones, occupational activities and long-distance trade of the Classic Maya from Xcambo--archaeological and osteological evidence. AB - Xcambo is a Classic period Maya site (250-700 AD) situated on the northern coast of Yucatan, Mexico. Archaeological evidence suggests that the site began as a salt production center but adopted a more administrative role as a commercial port in the Late Classic period. Economic growth, depending on its magnitude, could have affected the daily occupations of Xcambo's inhabitants. However, this is difficult to infer from the archaeological record. The aim of this study was to directly evaluate this possibility through skeletal analysis. Since diaphyseal robusticity and shape are predominantly influenced by mechanical loading history, long bone cross-sections can be used to access activity patterns. To this end, humeri and femora of 47 male and 35 female adult specimens from two Xcambo population samples were scrutinized. Our analysis satisfies general archaeological expectations and provides additional information on the population's physical response to economic growth. Decreasing robusticity and femoral anterior-posterior rigidity indicate an overall decrease in physical workload and mobility, concomitant with the site's increasing administrative function. We also observed a significant decrease in sexual dimorphism, possibly attributable to the differential response of male and female physical work spheres during socioeconomic change. In general, our findings suggest even nonsubsistence based socioeconomic change can significantly affect the bone structure of a population, rendering activity analysis an important aspect of the reconstruction of living conditions of past populations. PMID- 18383158 TI - Osteological and dental markers of health in the transition from the Late Antique to the Early Medieval period in Croatia. AB - The purpose of this study is to analyze health at the transition from the Late Antique (LA) to the Early Medieval (EM) period in Croatia. Results of the analyses of skeletal remains are compared with historical and archaeological data to test the hypothesis that the transition was catastrophic. An additional objective is to determine whether the transition was a uniform process, or differentially affected the past inhabitants of Croatia because of various local considerations. To accomplish this, four markers of health: cribra orbitalia, linear enamel hypoplasia, nonspecific periostitis, and trauma were compared in 981 skeletons: 477 from nine urban LA sites, and 504 from six rural EM sites. Data were collected by sex and age for individual, and for co-occurrences of various features. Because continental and Adriatic Croatia has different ecological features, data were specifically tabulated for the two regions. Comparisons between the continental and Adriatic regions of the LA series showed no significant differences in the frequencies of the analyzed markers of stress. Comparisons between the LA and EM series showed similar frequencies in continental Croatia--suggesting no significant discontinuity of living conditions, and a significant increase of cribra orbitalia, periostitis, and trauma frequencies during the EM period in Adriatic Croatia. The deterioration of living conditions primarily affected subadults and males. These data suggest that the transition from the LA to the EM period in Croatia was not a uniform process, but differentially affected population biology most likely because of local cultural, socio-economical or political considerations. PMID- 18383159 TI - Growth-related changes in prehistoric Jomon and modern Japanese mandibles with emphasis on cortical bone distribution. AB - Cortical bone distribution of the anthropoid mandibular symphysis has been addressed in relation to mechanical stress generated by mastication. To examine whether or not bone mass and distribution patterns of the human mandibular symphysis could be interpreted as an example of functional adaptation, we compared the skeletal growth series of two populations, prehistoric Jomon, considered to represent a "robust" mandibular morphology associated with a presumed heavier masticatory load, and modern Japanese. Results showed that the adult Jomon symphysis possessed significantly greater bone mass and thicker cortical bone compared to the modern Japanese condition. However, the second moments of area did not differ significantly between the two, indicating comparable rigidity against bending. Furthermore, the Jomon mandibles of the infant to juvenile stages exhibited most of the adult characteristics, in both bone mass/distribution of the symphysis and in mandibular corpus/ramus morphologies. The present study also demonstrated the presence of a growth pattern of symphyseal cortical thickness, common to both the Jomon and the modern Japanese series. In both populations, subsequent to deciduous molar occlusion, cortical bone tends to be thickest at the inferolingual symphysis, at the location where the highest tensile stresses presumably occur during mastication. These findings suggest that the "robust" characteristics of the Jomon mandible are initially manifested early in development, and that the effect of mechanical stimulus to bone mass formation in the human symphysis is largely confined to a regulatory role during growth modeling. PMID- 18383160 TI - Dental wear and oral pathology: possible evidence and consequences of habitual use of teeth in a Swedish Neolithic sample. AB - Atypical wear and oral lesions were studied in the dental remains from the Middle Neolithic Pitted Ware Culture site Ajvide on the island of Gotland in the Baltic Sea. Teeth (n = 764) from 17 males and 11 females were examined microscopically to document unusual wear, assess wear patterns and oral lesions as evidence of habitual behavior, and determine their effect on the oral health of this population. Five atypical wear types were observed: occlusal facets, occlusal excessive load, labial vertical striae, labial horizontal striae, and interproximal striae. Three oral lesions were recorded: chipping, periapical lesions, and dental tilting. Results indicate that teeth were used in a habitual manner at Ajvide, based on the morphology and regularity of the patterns of atypical wear. Differences were observed between the sexes, indicating gender related differences in the habitual use of teeth. Some wear categories showed a significant correlation with age, signifying increased or accumulated wear with age. Statistically significant positive correlations were found in the molars between occlusal excessive load wear and periapical lesions as well as tilting. Other apparent links were also observed between chipping and vertical striae as well as excessive load, although these were not significant. This suggests a relationship between dental wear and dental pathologies at this site, suggesting that habitual use of teeth indirectly affected the general oral health at the site. Wear patterns, furthermore, seem to mirror both frequent activities as well as single events. PMID- 18383161 TI - Protamine usage following implantation of drug-eluting stents: a word of caution. AB - The treatment of hemorrhagic complications of percutaneous interventions with protamine is an accepted practice. Concerns exist about the increased thrombo genicity of drug-eluting stents. We report two cases of acute stent thrombosis following drug-eluting stent implantation related to protamine administration. PMID- 18383162 TI - Transcatheter treatment for systemic-to-pulmonary artery shunt obstruction in infants and children. AB - Systemic-to-pulmonary artery shunt occlusion is a life-threatening complication in patients with shunt-dependent pulmonary blood flow. In the current era, the definitive diagnosis of shunt obstruction, and interventions to re-establish shunt patency are performed in the catheterization laboratory, rather than the operating room. This review summarizes the various transcatheter techniques used by the interventionalist to re-establish flow through occluded shunts. PMID- 18383163 TI - Bone-muscle interaction of the fractured femur. AB - The interaction forces of a fractured femur among the bone, muscle, and other soft tissues are not well understood. Only a small number of in vivo measurements have been made and with many limitations. Mathematical modeling is a useful alternative, overcoming limitations and allowing investigation of hypothetical simulated reductions. We aimed to develop a model to help understand best practices in fracture reduction and to form a base to develop new technologies and procedures. The simulation environment allows muscle forces and moments to deform a fractured femur, and the behavior of forces during reduction can be found. Visual and numerical output of forces and moments during simulated reduction procedures are provided. The output can be probed throughout the reduction procedure down to the individual muscle's contribution. This is achieved by construction of an anatomically correct three-dimensional mathematical model of the lower extremity and muscles. Parameters are fully customizable and can be used to investigate simple, oblique, and some comminuted fractures. Results were compared with published in vivo measurements and were of the same magnitude. A simple midshaft fracture had a maximum resulting force of 428 N, whereas traction from the hip reached a maximum value of 893 N at 60 mm of displacement. Monte Carlo analysis revealed that the deforming force was most sensitive to the muscles' rest lengths. The developed model provides greater understanding and detail than in vivo measurements have to date. It allows new treatment procedures to be developed and importantly to assess the outcome. PMID- 18383164 TI - Feasibility of primary clot extraction prior to percutaneous coronary intervention in acute myocardial infarction. AB - OBJECTIVES: To test the feasibility, safety, and in-hospital outcomes of utilizing the FilterWire EZ to extract clot prior to percutaneous coronary intervention (PCI) in patients presenting with acute myocardial infarction (MI). BACKGROUND: PCI in patients with acute MI is associated with a higher incidence of distal embolization, no-reflow, or slow flow partly due to the presence of clot burden. METHODS: The authors describe the feasibility, safety, and outcomes of using a FilterWire EZ distal protection device as a clot extraction device in patients who presented with acute MI and documented clot on coronary angiography. RESULTS: Fifteen consecutive male patients with a mean age of 54 +/- 8 years presented with acute MI (60% ST elevation MI). MI involved left anterior descending artery (n = 4), circumflex artery (n = 3), and right coronary artery (n = 8). Clot extraction followed by PCI reduced the percent diameter stenosis from 94 +/- 12 to 65 +/- 11 (P < 0.001) and restored TIMI 3 flow in all patients without distal embolization. The angiographic, procedural, and clinical success rates were 100%. The mean left ventricular ejection fraction (LVEF) was 52 +/- 8% (range 30-62%) with only three patients (15%) who had an LVEF <50% and five patients (33%) without apparent wall motion abnormalities on echocardiography. CONCLUSIONS: Clot extraction before PCI during acute MI in native coronaries is feasible, safe, and effective in restoring TIMI 3 flow without distal embolization. Whether this approach results in better outcomes and improved LV function compared with standard therapy alone requires further investigation. PMID- 18383165 TI - Treatment of uncomplicated vaso-occlusive crises in children with sickle cell disease in a day hospital. AB - BACKGROUND: Day hospital management for patients with sickle cell disease experiencing uncomplicated vaso-occlusive pain crises has been described in adult populations as an alternative care delivery system. The objective of this study was to characterize and descriptively assess the benefits of a day hospital exclusively designed for children. PROCEDURE: We retrospectively studied all admissions to the Day Hospital at the Texas Children's Sickle Cell Center since its inception in 2000. A Day Hospital admission was defined as a minimum of two consecutive days of aggressive pain management as an outpatient, including intravenous hydration and analgesics, supported by home treatment over night with oral analgesic and anti-inflammatory agents. We gathered data on demographics, incoming pain score, provider type, opioid administration, length of stay, and needs for higher level care. RESULTS: A total of 35 patients, ages 2-19, accounted for 80 episodes during the study period. The median incoming pain score was 8 on a scale from 1 to 10. The median length of stay was 2 days. The return rate for acute care within 48 hr for persistent symptoms was 7%. Seventy-one percent of patients admitted to the Day Hospital were treated without requiring transfer to inpatient care for escalating pain or medical needs. CONCLUSIONS: We conclude that a dedicated Day Hospital facility has the potential to provide patient-centered, effective, and timely management of vaso-occlusive crises in children as well as adults. PMID- 18383166 TI - Carotid sheath rescue with a distal filter retrieval catheter. AB - Sheath placement prior to carotid artery stenting is usually uncomplicated, and provides sufficient support for the procedure. In certain patients, especially those with unfavorable arch anatomy, tortuous vessels, and heavily calcified lesions, the sheath backs out into the aortic arch with compromise of wire and embolic protection device (EPD) position, and risk of "dragging" the EPD back through the lesion. A novel use of the distal filter retrieval catheter to "rescue" a prolapsed guide sheath is described. Use of the filter retrieval catheter as a "body" to retrack the sheath but not recapturing the deployed filter is a useful technique, since the equipment is already available. This avoided the need to pull a retrieved filter through a severe undilated carotid stenosis, reducing the amount of manipulations needed to reposition the sheath and thus reducing the risk of embolic events. PMID- 18383167 TI - Novel use of a modified Amplatzer Vascular Plug to occlude a patent ductus arteriosus in two patients. AB - We describe two cases in which a patent ductus arteriosus (PDA) was successfully occluded percutaneously, using the novel approach of prefilling an Amplatzer Vascular Plug with embolization coils to decrease the risk of residual shunting through the device. One patient was a small premature neonate, and the other was a child with an aneurismal, tubular PDA with no aortic ampulla. These are situations in which more popular PDA devices such as the Amplatzer Duct Occluder may be contraindicated. PMID- 18383168 TI - Effect of quercetin on bone formation. AB - Quercetin is a flavonol, also a phytoestrogen, available commonly in onions and apples. Our laboratory compared the amount of new bone produced by quercetin in a collagen matrix to that produced by bone grafts and a collagen matrix. Eighteen bone defects, 5 x 10 mm were created in the parietal bone of nine New Zealand White rabbits. In the experimental group, six defects were grafted with quercetin solution mixed with a collagen matrix. In the control groups, six defects were grafted with a collagen matrix alone (active control) and six were left empty (passive control). Animals were killed on day 14 and the defects were dissected and prepared for histological assessment. Serial sections were cut across each defect. Quantitative analysis of new bone formation was made on 100 sections (50 sections for each group) using image analysis. A total of 556% more new bone was present in defects grafted with quercetin in a collagen matrix than those grafted with a collagen matrix alone. No bone was formed in the passive control group. In conclusion, quercetin in a collagen matrix has the effect of increasing new bone formation locally, and can be used as a bone graft material. PMID- 18383169 TI - Catheter-based treatment of the subclavian and innominate arteries. AB - OBJECTIVES: We report outcomes in patients undergoing catheter-based intervention for symptomatic subclavian and innominate artery (S/IA) atherosclerosis. BACKGROUND: Symptomatic S/IA obstructive lesions have traditionally been treated with open surgical revascularization. Catheter-based endovascular therapies reduce the morbidity and mortality associated with surgery in many vascular beds. METHODS: Between December 1993 and May 2006, 170 patients underwent primary stent placement in 177 S/IA arteries. Indications for revascularization included arm ischemia (57%), subclavian steal syndrome (37%), coronary-subclavian steal syndrome (21%), and planned coronary bypass surgery with the involved internal mammary artery (8%). RESULTS: Technical success was achieved in 98.3% (174/177) arteries, including 99.4% for stenotic lesions (155/156) and 90.5% for occlusions (19/21). There were no procedure-related deaths and one stroke (0.6%, 1/170). Follow-up was obtained in 151 (89%) patients at 35.2 +/- 30.8 months, with a target vessel revascularization rate of 14.6% (23/157). At last follow-up, 82% (124/151) of all treated patients remained asymptomatic with a primary patency of 83% and a secondary patency of 96%. CONCLUSIONS: Catheter-based revascularization with stents for symptomatic S/IA lesions is safe and effective with excellent patency rates and sustained symptom resolution in the majority (>80%) of patients over 3 years of follow-up. Percutaneous primary stent therapy is the preferred method of revascularization in patients with suitable anatomy. PMID- 18383170 TI - Comparison of five-year outcome in African Americans versus Caucasians following percutaneous coronary intervention. AB - BACKGROUND: Studies regarding short-term outcomes after percutaneous coronary intervention (PCI) have reported no ethnic differences and data on long-term follow-up is conflicting and sparse. METHODS: 730 consecutive patients (67% African American) undergoing PCI from January 1999 to December 2000 at a tertiary care center in Detroit, MI, were followed up. End points studied included either all cause mortality collected from Social Security Death Index or first hospital admission after the index procedure due to myocardial infarction(MI), congestive heart failure(CHF), and revascularization (PCI or coronary artery bypass graft surgery). RESULTS: African-Americans undergoing PCI had significant differences in baseline cardiovascular co-morbidity and were more likely to present with acute myocardial infarction than Caucasians. On Kaplan Meier survival analysis and log rank test, each ethnic group had equivalent survival for cumulative end points upto 6-month follow-up, however longer follow-up to 5 year was characterized by lower survival rate in African Americans compared to Caucasians (41% vs. 54%, log rank P 0.01). After adjustment for potential confounders, AA ethnicity (Adjusted HR 1.62, 95% CI 1.01-1.28, P 0.04) remained a predictor of adverse cardiac outcome (Death/MI/CHF) at five-year follow-up (Cox regression propensity adjusted hazard analysis). CONCLUSIONS: African American patients undergoing PCI had unfavorable baseline cardiovascular characteristics but comparable short-term outcome compared to whites. However, at 5-year follow-up, African Americans had worse clinical outcome, higher incidence of acute myocardial infarction, congestive heart failure and significantly lower long-term survival. PMID- 18383171 TI - Thermal damage potential during hip resurfacing in osteonecrosis of the femoral head: an experimental study. AB - Hip resurfacing arthroplasty has become an attractive treatment option for young, active patients with femoral head necrosis. However, little information is available about the potential thermal damage to the remaining femoral head when a cemented component is used. We used an experimental model to measure the temperature profile at the cement-bone interface during hip resurfacing. We compared four simulated lesion sizes-15, 25, 33, and 50%-of the femoral head, and a control group with no cystic lesion. Temperatures were measured with the specimens in a 37 degrees C saline bath or with copious pulsed lavage. With specimens tested in the bath, peak temperatures were higher, and durations of temperatures above 50 degrees C were longer, in femoral heads with necrotic lesions (88.8 +/- 7.5 degrees C; 17.6 +/- 1.1 min for a 15% lesion; 96.2 +/- 7.2 degrees C; 22.86 +/- 1.3 min for a 25% lesion; 99.7 (c) 200 +/- 9.4 degrees C; 28.6 +/- 2.0 min for a 33% lesion; and 97.2 +/- 4.2 degrees C; 35.6 +/- 2.4 min for a 50% lesion) than those in the control group (65.8 +/- 4.9 degrees C; 10.0 +/- 1.3 min). The larger the cement-filled cysts, the longer the temperatures remained above 50 degrees C. Although copious lavage reduced the temperature profile in each group, the temperatures remained above 50 degrees C for 7 to 17 min in specimens with necrotic cysts. The measured temperatures during surface replacement are sufficiently high in magnitude and long in duration to cause thermal damage to the remaining bone in femoral heads with preexisting necrotic lesions. Hip resurfacing for femoral head necrosis should be performed with caution. PMID- 18383172 TI - Percutaneous removal of an embolized port catheter: description of a new coaxial recovery technique including a case-report. AB - The achievement of coaxiality between recovery device and embolized tubular foreign bodies like central venous catheters is a key point to achieve percutaneous removal. To facilitate this target, we designed a coaxial retrieval loop-snare device using a modified coronary guiding catheter and a coronary 0.014'' guidewire. This new technique has been bench tested and then successfully used to remove a long Port-A catheter's fragment embolized into the right sided heart. PMID- 18383173 TI - Dorsalis pedis artery stenting for limb salvage. AB - Stent-supported angioplasty has emerged as an effective alternative to distal bypass for infrapopliteal artery disease. This treatment has been mainly employed in the tibial and peroneal arteries. However, the pedal artery below the ankle is the final frontier for catheter intervention. Here, we describe the utility of dorsalis pedis artery stenting for limb salvage. PMID- 18383174 TI - Percutaneous transthoracic ventricular puncture for diagnostic and interventional catheterization. AB - OBJECTIVE: To describe our experience in a case series of patients requiring percutaneous direct ventricular puncture and sheath placement for diagnosis or intervention. BACKGROUND: Access to the right or left ventricle for percutaneous interventions is limited in patients with mechanical prostheses in either the tricuspid, or mitral and aortic positions. METHODS: After coronary angiography, direct ventricular puncture under ultrasound and fluoroscopic guidance was performed. At end of case, protamine was given to reverse the heparin, and sheaths were pulled with purse-string suture closure of the skin entrance. RESULTS: For right ventricular access, 8- to 9-F sheaths were placed from subxiphoid approach in 2 patients to allow conduit and pulmonary artery interventions. For left ventricular access in patients with mitral and aortic prostheses, 4- to 8-F sheaths were placed from apical approach to allow diagnostic evaluation in 1 and interventions in 5 to occlude perivalvular mitral leaks and postoperative ventricular septal defect. Complication in one consisted of intercostal vein injury resulting in hemothorax requiring chest tube drainage. CONCLUSION: In this small cases series, direct ventricular puncture allowed the intervention to proceed with up to 9-F sheath size. Attention to puncture site relative to intercostal vascular anatomy is warranted. PMID- 18383175 TI - Risk of nonmelanoma skin cancer with azathioprine use. AB - While often life-saving for many complex diseases, iatrogenic immunosuppression has been associated with life-threatening infections and malignancies. Among these malignancies is skin cancer. Skin cancer is the most common form of cancer in the United States; the nonmelanoma skin cancers have an annual incidence of greater than 1,000,000 people in the US. It is well documented that the risk of nonmelanoma skin cancer (NMSC) is increased in those who are immunosuppressed. While many articles have been published on skin cancer risk in organ transplant recipients, little has been written regarding the incidence of nonmelanoma skin cancer in inflammatory bowel disease. A review of the literature of patients who are immunosuppressed for autoimmune disorders, and specifically, inflammatory bowel diseases, is discussed, as well as clinical presentations and treatment options. PMID- 18383176 TI - Clinical peer review: a plea for fairness. PMID- 18383177 TI - Guttate psoriasis induced by infliximab in a child with Crohn's disease. PMID- 18383178 TI - Anti-TNFalpha therapy in a rare case of Crohn's disease: still a panacea. PMID- 18383179 TI - NOD2/CARD15 genotype influences MDP-induced cytokine release and basal IL-12p40 levels in primary isolated peripheral blood monocytes. AB - BACKGROUND: The functional link between mutations in NOD2 and Crohn's disease (CD) has not been entirely elucidated. The 1007fs mutation results in loss of NF kappaB activation in response to muramyl dipeptide (MDP) but has also been linked to an increased IL-1beta processing and IL-12 release. METHODS: We investigated the basal and MDP-triggered mRNA expression and protein release for TNF-alpha, IL 10, IL-1beta, and IL-12p40 in peripheral blood monocytes from 40 CD patients and 15 healthy individuals with different NOD2 genotypes. RESULTS: Monocytes from individuals with 2 mutated NOD2 alleles (homozygous and compound-heterozygous individuals) displayed an impaired release of TNF-alpha and IL-10 but also of IL 1beta and IL-12p40 in response to MDP. In contrast to other NOD2 variants, the presence of at least 1 1007fs allele in double-mutated individuals completely abrogated NOD2 receptor function. Interestingly, monocytes from CD patients with 2 mutated NOD2 alleles displayed significantly higher basal levels of IL-12p40 in cell culture supernatants compared to wildtype CD patients and control individuals (P = 0.002 and P = 0.008, respectively). This was regardless of the IL23R genotype and was not mirrored by increased IL-12p40 plasma levels in these individuals. CONCLUSIONS: The CD-associated NOD2 variants lead, in a dose- and mutation-dependent manner, to an impaired release of TNF-alpha, IL-10, IL-1beta, and IL-12p40 in response to MDP. The finding of increased basal levels for IL 12p40-related cytokines in monocytes with 2 mutated NOD2 alleles is likely to set a new link between NOD2 mutations and the inflammatory mechanisms underlying CD. PMID- 18383180 TI - Hybrid balloon pulmonary valvuloplasty in a 700-g infant: Thinking outside the box. AB - This report describes a unique Hybrid approach to balloon pulmonary valvuloplasty in a 700-g premature infant with pulmonary valve stenosis and very poor right ventricular function. PMID- 18383181 TI - Gender differences in passive knee biomechanical properties in tibial rotation. AB - The anterior cruciate ligament (ACL) is the most commonly injured knee ligament with the highest incidence of injury in female athletes who participate in pivoting sports. Noncontact ACL injuries commonly occur with both internal and external tibial rotation. ACL impingement against the lateral wall of the intercondylar notch during tibial external rotation and abduction has been proposed as an injury mechanism, but few studies have evaluated in vivo gender specific differences in laxity and stiffness in external and internal tibial rotations. The purpose of this study was to evaluate these differences. The knees of 10 male and 10 female healthy subjects were rotated between internal and external tibial rotation with the knee at 60 degrees of flexion. Joint laxity, stiffness, and energy loss were compared between male and female subjects. Women had higher laxity (p = 0.01), lower stiffness (p = 0.038), and higher energy loss (p = 0.008) in external tibial rotation than did men. The results suggest that women may be at greater risk of ACL injury resulting from impingement against the lateral wall of the intercondylar notch, which has been shown to be associated with external tibial rotation and abduction. PMID- 18383182 TI - The effect of wrist position on the relative motion of tendon, nerve, and subsynovial connective tissue within the carpal tunnel in a human cadaver model. AB - The purpose of this study was to measure the effect of wrist position on the relative motion of the middle finger flexor digitorum superficialis (FDS) tendon, subsynovial connective tissue (SSCT), median nerve, and flexor retinaculum during simulated active finger motion. The relative motion of each tissue was measured by fluoroscopy in 10 human cadavers. Measurements were obtained for wrist positions of neutral (0 degree extension), 30 and 60 degrees of flexion, and 30 and 60 degrees of extension. The shear strain index (SSI) was defined as the difference in motion between two tissues (tendon, SSCT, or nerve) divided by tendon excursion, expressed as a percentage. The motion of the tendon, SSCT, and nerve in the 60 degree flexed position was significantly less than the motion in all other wrist positions (p < 0.001). The SSI at 60 degrees of flexion for tendon-SSCT and tendon-nerve were significantly increased compared with all other positions (p < 0.001). Because the SSCT and tendon are physically connected, a decrease in SSCT motion relative to the tendon would increase the shear strain on the SSCT with tendon motion. Thus, this result suggests that the SSCT may be predisposed to shear injury from activity done in 60 degrees of wrist flexion. PMID- 18383184 TI - Inference from genome-wide association studies using a novel Markov model. AB - In this paper we propose a Bayesian modeling approach to the analysis of genome wide association studies based on single nucleotide polymorphism (SNP) data. Our latent seed model combines various aspects of k-means clustering, hidden Markov models (HMMs) and logistic regression into a fully Bayesian model. It is fitted using the Markov chain Monte Carlo stochastic simulation method, with Metropolis Hastings update steps. The approach is flexible, both in allowing different types of genetic models, and because it can be easily extended while remaining computationally feasible due to the use of fast algorithms for HMMs. It allows for inference primarily on the location of the causal locus and also on other parameters of interest. The latent seed model is used here to analyze three data sets, using both synthetic and real disease phenotypes with real SNP data, and shows promising results. Our method is able to correctly identify the causal locus in examples where single SNP analysis is both successful and unsuccessful at identifying the causal SNP. PMID- 18383183 TI - Synthesis and structure of built-up organic macromolecules containing helicene. AB - Built-up macromolecules are acyclic molecules with molecular weights of several thousand daltons, which are synthesized by connecting small molecular units using stepwise methods. The chemical study of built-up macromolecules reveals some noteworthy properties that are different from those of conventional biological and synthetic macromolecules. A characteristic feature of built-up organic macromolecules is that their structures and properties are discontinuous at a certain molecular weight. For such macromolecules, variation in the small molecular units and the formation of cyclic structures substantially affect the structure and properties. The built-up organic macromolecules obtained by connecting helicenes with amide, acetylene, and amine groups are discussed in this paper. Some chiral built-up macromolecules are linked by covalent bonds, and the effects of linking on the structure are compared. PMID- 18383185 TI - Hyaluronic acid and chondroitin sulfate content of osteoarthritic human knee cartilage: site-specific correlation with weight-bearing force based on femorotibial angle measurement. AB - This study analyzed glycosaminoglycan (GAG) content in specific compartments of the knee joint to determine the impact of malalignment and helped refine indications for osteotomy. To assess malalignment, the radiological femorotibial angle (FTA) was measured and knee joints were also graded for OA severity with the Kellgren/Lawrence (K/L) classification. Cartilage samples were obtained from 36 knees of 32 OA patients undergoing total knee replacement surgery. Explants were harvested from the medial femoral condyle (MFC), lateral femoral condyle (LFC), patellar groove (PG), and lateral posterior femoral condyle (LPC). Concentrations of hyaluronic acid (HA) and chondroitin sulfate (CS) were measured by high-performance liquid chromatography (HPLC). With OA severity, the average FTA significantly increased. HA and CS content in MFC was negatively correlated with radiographic FTA. In LFC, HA ratio, which is HA content in lateral condyle divided by medial condyle and chondroitin 6 sulfate, increased until about 190 degrees FTA. Importantly, at >190 degrees these contents were significantly decreased. HA and CS content of the femoral condyle shows topographic differences that are related to OA grade and weight-bearing force based on FTA. The clinical relevance is that osteotomy may not be indicated for patients with severe varus (>190 degrees) abnormalities. PMID- 18383186 TI - Combination of rituximab with chemotherapy in diffuse large B-cell lymphoma. Evaluation in daily practice before and after approval of rituximab in this indication. AB - Randomized trials have demonstrated improved outcome from adding rituximab to CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) for patients with diffuse large B-cell lymphoma (DLBCL). This retrospective study compared the outcomes of 224 patients with DLBCL treated in our institution before (Period 1, 1996-2002) and after (Period 2, 2002-2005) approval of rituximab in this indication to evaluate the impact of the drug in daily practice in unselected patients receiving different types of chemotherapy. We treated 131 patients in Period 1 versus 93 in Period 2 (median follow-up, 75 and 29 months, respectively) with no difference in patient characteristics between the two periods. Event-free and overall survivals (EFS and OS) were significantly improved in Period 2 for elderly patients and a significant shift in the selection of regimens was observed at the time when rituximab became available. More patients received the CHOP regimen in Period 2 than in Period 1 (82 vs. 57%, p < 0.007) with CHOP being substituted for epirubicin-based regimens. In younger patients treated mostly with the ACVBP regimen (doxorubicin, cyclophosphamide, vindesine, bleomycin, prednisone) these differences were not observed, suggesting that combination of rituximab with dose-dense chemotherapy may deserve further evaluation in this age group. PMID- 18383187 TI - Acute myeloid leukaemia after treatment with (90)Y-ibritumomab tiuxetan for follicular lymphoma. AB - We report here a patient with relapsed follicular lymphoma who developed secondary acute myeloid leukaemia 15 months after radioimmunotherapy (RIT) with (90)Y-ibritumomab tiuxetan, the fifth described case to date. We review the literature for the potential causal relationship between RIT and secondary myelodysplastic syndromes/acute myeloid leukaemia. PMID- 18383188 TI - Abuse by marriage: the exploitation of mentally ill older people. AB - OBJECTIVES: (i) To raise awareness about the vulnerability of mentally ill older persons to abuse by others seeking to gain by marriage; (ii) to outline key legal cases from common law countries; and (iii) to provide guidelines for health care professionals who encounter this issue in practice. METHOD: We present two cases: the first case involved an 87-year-old widower who married his carer--50 years his junior--in a religious ceremony while hypomanic. The second case involved an 82-year-old widow with moderate dementia who married her boarder, the marriage subsequently being found void in the Family Court of Australia on the basis that her consent was not real because she was incapable of understanding the nature and effect of the marriage ceremony. RESULTS: Abuse by marriage may be of a psychological, sexual, social or financial nature.Older people with impaired judgement and inability to appraise others due to mental illness may be persuaded to execute legal documents such as marriage certificates. CONCLUSION: Health care professionals may have a role in the identification and management of this kind of abuse. There are legal means to address this problem ranging from guardianship and financial management to family law court applications to seek a decree of nullity/invalidity of the marriage. PMID- 18383189 TI - Coping strategies, anxiety and depression in caregivers of people with Alzheimer's disease. AB - OBJECTIVES: There have been few longitudinal studies investigating the impact of coping on psychological morbidity in caregivers of people with dementia. Findings have been inconsistent and little attention has been paid to anxiety in caregivers. We explored the hypothesis that the relationship between caregiver burden and anxiety and depression is mediated by coping style. METHODS: As part of the LASER-AD study, 93 (73.8%) people with Alzheimer's disease and their family caregivers recruited at baseline were re-interviewed 1 year later. Sampling aimed to ensure that the participants were representative of people living in the UK with Alzheimer's disease in terms of dementia severity, gender and care setting. We used the Hospital Anxiety and Depression Scale, the Zarit Burden scale and the Brief COPE to measure coping strategies. RESULTS: Using fewer emotion-focused strategies and more problem-focused strategies (but not dysfunctional strategies) mediated the relationship between caregiver burden and anxiety a year later, after controlling for potential confounders, in a well fitting structural equation model (chi(2)=0.93, df=3, p=0.82; NFI=1.0, RFI=0.97, IFI=1.0, TLI=1.1, RMSEA=0.0). Using fewer emotion focused strategies also predicted higher psychological morbidity in general. The hypothesised relationship was not proved for depression. CONCLUSIONS: Using emotion-focussed coping strategies in response to caregiver burden seemed to protect caregivers from developing higher anxiety levels a year later; however using problem focussed strategies did not. Our results suggest that a psychological intervention package to emphasise emotion-focused coping may be a rational approach to reduce anxiety in dementia caregivers. Studies are needed to test such interventions. PMID- 18383190 TI - Sheathless focusing of microbeads and blood cells based on hydrophoresis. AB - This paper presents a microfluidic device for sheathless focusing of microbeads and blood cells based on a hydrophoretic platform comprising a V-shaped obstacle array (VOA). The VOA generates lateral pressure gradients that induce helical recirculations. Following the focusing flow particles passing through the VOA are focused in the center of the channel. In the device, the focusing pattern can be modulated by varying the gap height of the VOA. To achieve complete focusing within 4.4% coefficient of variation, the relative size differences between the gap and the particle were 3 and 4 microm for 10 and 15 microm beads, respectively. Red blood cells were used to study the hydrophoretic focusing pattern of biconcave, disk-shaped particles. PMID- 18383191 TI - Doping-dependent electrical characteristics of SnO2 nanowires. PMID- 18383192 TI - A single-walled-carbon-nanotube filter for removal of viral and bacterial pathogens. PMID- 18383193 TI - Nanowires enabling signal-enhanced nanoscale Raman spectroscopy. AB - Silicon nanowires grown by the vapor-liquid-solid (VLS) mechanism catalyzed by gold show gold caps (droplets) approximately 20-500 nm in diameter with a half spherical towards almost spherical shape. These gold droplets are well suited to exploit the surface-enhanced Raman scattering (SERS) effect and could be used for tip-enhanced Raman spectroscopy (TERS). The gold droplet of a nanowire attached to an atomic force microscopy (AFM) tip could locally enhance the Raman signal and increase the spatial resolution. Used as a SERS template, an ensemble of self organizing nanowires grown bottom up on a silicon substrate could allow highly sensitive signal-enhanced Raman spectroscopy of materials that show a characteristic Raman signature. A combination of a nanowire-based TERS probe and a nanowire-based SERS substrate promises optimized signal enhancement so that the detection of highly dilute species, even single molecules or single bacteria or DNA strands, and other soft matter is within reach. Potential applications of this novel nanowire-based SERS and TERS solution lie in the fields of biomedical and life sciences, as well as security and solid-state research such as silicon technology. PMID- 18383194 TI - Informing the selection of futility stopping thresholds: case study from a late phase clinical trial. AB - In an environment where (i) potential risks to subjects participating in clinical studies need to be managed carefully, (ii) trial costs are increasing, and (iii) there are limited research resources available, it is necessary to prioritize research projects and sometimes re-prioritize if early indications suggest that a trial has low probability of success. Futility designs allow this re prioritization to take place. This paper reviews a number of possible futility methods available and presents a case study from a late-phase study of an HIV therapeutic, which utilized conditional power-based stopping thresholds. The two most challenging aspects of incorporating a futility interim analysis into a trial design are the selection of optimal stopping thresholds and the timing of the analysis, both of which require the balancing of various risks. The paper outlines a number of graphical aids that proved useful in explaining the statistical risks involved to the study team. Further, the paper outlines a decision analysis undertaken which combined expectations of drug performance with conditional power calculations in order to produce probabilities of different interim and final outcomes, and which ultimately led to the selection of the final stopping thresholds. PMID- 18383195 TI - Exenatide-induced depression in a geriatric patient. PMID- 18383196 TI - Efficacy of five selected acaricides against Tetranychus urticae (Acari: Tetranychidae) and their side effects on relevant natural enemies occurring in citrus orchards. AB - BACKGROUND: Three groups of natural enemies are fundamental in citrus IPM in Spain: coccinellid and phytoseiid predators and hymenopteran parasitoids. Tetranychus urticae Koch is an important pest affecting citrus, for which biological control has not yet been achieved; therefore, acaricides are commonly used to control it. The goal of this study was to measure the efficacy of different acaricides on this mite and their side effects on three natural enemies relevant for citrus (Cryptolaemus montrouzieri Mulsant, Neoseiulus californicus McGregor and Aphidius colemani Viereck). RESULTS: Some products proved highly effective against T. urticae and harmless to A. colemani (mineral oil, tebufenpyrad, clofentezine and fenazaquin). However, almost all products tested were slightly harmful for both the predators considered. Fenazaquin was even moderately harmful for N. californicus. CONCLUSION: Further studies, like that presented here, are necessary to gain a better understanding of integrating biological and chemical controls. When considering both efficacy and side effects on beneficial arthropods, the best options would seem to be mineral oil, tebufenpyrad and clofentezine. However, it is urgent to complete testing of the side effects of the acaricides used in citrus. This question is crucial if the fact that two recently introduced Tetranychidae are being controlled in citrus by chemical means exclusively is considered. PMID- 18383197 TI - Cross-resistance between a Bacillus thuringiensis Cry toxin and non-Bt insecticides in the diamondback moth. AB - BACKGROUND: Bacillus thuringiensis Berliner (Bt) crystal (Cry) toxins are expressed in various transgenic crops and are also used as sprays in integrated pest management and organic agricultural systems. The diamondback moth (Plutella xylostella L.) is a major worldwide pest of crucifer crops and one that has readily acquired field resistance to a broad range of insecticides. RESULTS: Selection of a subpopulation of the P. xylostella SERD4 population with the pyrethroid deltamethrin increased resistance to both deltamethrin and Cry1Ac relative to an unselected subpopulation. Selection of a second subpopulation with the Bt toxin Cry1Ac also increased resistance to both Cry1Ac and deltamethrin. A complementation test between the Cry1Ac-selected and deltamethrin-selected subpopulations suggested the presence of a common genetic locus or loci that control resistance to both insecticides. A piperonyl butoxide analogue with potent inhibitory activity against insect esterases significantly increased the toxicity of Cry1Ac and deltamethrin against the respective resistant subpopulations, but showed no such synergism with the unselected subpopulation of SERD4. CONCLUSION: Selection of one resistance phenotype resulted in the simultaneous selection of the other. This phenomenon could be due to a single mechanism acting against both classes of insecticide or to genetically linked, but separate, mechanisms. PMID- 18383198 TI - A combined drama-based and CBT approach to working with self-reported anger aggression. AB - BACKGROUND: A drama-based programme, called 'Insult to Injury', was designed to explore the processes of anger, aggression and violence. The aim of the programme was to enable offenders to identify and generate strategies and skills for dealing with potentially volatile situations, and to provide a safe and supportive environment in which to practice and evaluate these strategies. AIMS An active drama-based approach combined with cognitive-behavioural techniques was used to explore issues such as masculinity, power and control, pride and shame and victim awareness. Reductions in anger were hypothesized. METHOD A single group pre/post design assessed the levels of anger before and after the course. RESULTS Sixty-two adult male offenders from six prison establishments in the UK took part in the nine-day course. As hypothesised, significant reductions in anger were found in pre- to post-course assessment. CONCLUSIONS: These results suggest that a drama-based approach may be a promising adjunct to traditional anger management programmes for violent offenders. PMID- 18383199 TI - Are psychiatrists affecting the legal process by answering legal questions? AB - BACKGROUND: Psychiatrists are often asked to answer legal questions. The extent to which they answer strictly legal rather than medical matters is not known. AIM: To investigate how strongly psychiatrists in England and Wales express opinions on one legal question - that of diminished responsibility in respect of a murder charge, and how this is related to outcome in court. METHOD Our data were extracted from psychiatric reports and case files supplied by the then Department of Constitutional Affairs (now the Ministry of Justice) on cases heard in the Crown Courts between 1 January 1997 and 31 December 2001 in which the defence of diminished responsibility had been raised. The cases had been selected by the Law Commission in their earlier review of partial defences to murder. We devised a reliable system of rating the presence/absence and strength of expression of a legal opinion in the medical reports. We tested the data for relationship between nature and strength of opinion and progression to trial and verdict. RESULTS Psychiatric reports were available on 143 of 156 cases in which diminished responsibility was considered. They yielded 338 opinions on at least one aspect of diminished responsibility. In 110 (93%) of the 118 cases in which there was a diminished verdict, this was made without trial and, therefore, without reference to a jury. In only eight (27%) out of the 30 cases that went to trial, was a diminished responsibility verdict made. Half of the reports (169) gave a clear opinion on diminished responsibility, a third (121) invited the court to draw a particular conclusion and only 11% (36) provided relevant evidence without answering the legal questions. When there was an opinion or an invitation to make a finding on the legal question, a trial was less likely. A trial was also less likely if reports agreed on what the verdict should be. CONCLUSIONS Psychiatrists frequently answer the legal question of diminished responsibility. The judiciary and medical experts should join in research to examine the consequences of different styles or approaches in presentation of essentially similar evidence in court. PMID- 18383200 TI - Specificity of cognitive distortions to antisocial behaviours. AB - INTRODUCTION/BACKGROUND: Cognitive distortions have long been posited to facilitate antisocial behaviours, but the specificity of such distortions has rarely been studied. AIMS: To replicate findings of specificity between particular cognitions and externalizing or internalizing behaviours; to test for specificity of relationship between particular cognitions and different types of externalizing behaviours. METHODS: The participants were 239 male youths aged 10 to 19 years (mean (M) = 14.22, standard deviation (SD) = 1.64) from schools on the island of Curacao. Their cognitive distortions and problem behaviours were investigated through self-report. Results In controlled analyses, self-serving cognitive distortions were associated with externalizing behaviours whereas self debasing cognitive distortions were associated with internalizing behaviours. Within the externalizing domain, self-serving distortions with overt behavioural referents were linked to aggressive behaviour while self-serving distortions with covert behavioural referents were linked to delinquent behaviour. Within the aggression domain, distortions with opposition-defiance referents related to verbal aggression whereas distortions with physical aggression referents related to physically aggressive behaviour. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: The degree of cognitive-behavioural specificity documented by this study was remarkable. The observed pattern suggests that cognitive interventions designed for externalizing versus internalizing behaviours should differ in therapeutic approach. PMID- 18383201 TI - Homelessness in the state and federal prison population. AB - AIMS: This study sought to investigate the rates and correlates of homelessness (i.e. living on the street or in a homeless shelter), including mental illness, among US adult state and federal prison inmates (ASFPIs). METHOD: Data from a national survey of ASFPIs based on a random sampling survey (N = 17,565) were used to compare the homelessness rate among AFSPIs with that in the general population. Logistic regression was then used to examine the association of homelessness among ASFPIs with factors including symptoms, treatment of mental illness, previous criminal justice involvement, specific crimes, and demographic characteristics. RESULTS: Nine percent of ASFPIs reported an episode of homelessness in the year prior to arrest, 4-6 times the estimated rate in the general US adult population after allowing for age, race/ethnicity, and gender. In comparison to other inmates, these homeless inmates were more likely to be currently incarcerated for a property crime, but also to have had previous criminal justice system involvement for both property and violent crimes, to suffer from mental health and/or substance abuse problems, and to be more likely to have been unemployed and with a low income. CONCLUSIONS: Recent homelessness is far more common among ASFPIs than the general population. Prior incarceration, mental illness, substance abuse and disadvantageous socio-demographic characteristics were all found to be associated with homelessness among prison inmates, suggesting that there are several important factors in addition to efforts to survive with limited resources through criminal acts that influence the rates of homelessness among incarcerated individuals. PMID- 18383202 TI - The treatment of paraphilias: an historical perspective. PMID- 18383203 TI - A 'trans-transdiagnostic' model of the eating disorders: a new way to open the egg? PMID- 18383205 TI - An a posteriori calibration method for improving protein identification accuracy in proteomics using electrospray ionization time-of-flight tandem mass spectrometry. PMID- 18383206 TI - A generic method to detect electrophilic intermediates using isotopic pattern triggered data-dependent high-resolution accurate mass spectrometry. AB - A need still exists for a liquid chromatography/tandem mass spectrometry (LC/MS/MS) method that can detect broad classes of glutathione (GSH) conjugates and provide characterization of their structures. We now describe the development of a method that multiplexes high-resolution accurate mass analysis with isotope pattern triggered data-dependent product ion scans, for simultaneous detection and structural elucidation of GSH conjugates within a single analysis using a LTQ/Orbitrap. This method was initially developed to detect GSH conjugates generated from incubating 10 microM test compound with pooled human liver microsomes fortified with NADPH-regenerating system and a 2:1 ratio of 5 mM glutathione and [(13)C(2) (15)N-Gly]glutathione. The GSH conjugates were detected by isotope search of mass defect filtered and control subtracted full scan accurate MS data using MetWorks software. This was followed by elucidation of reactive intermediate structures using chemical formulae for both protonated molecules and their product ions from accurate masses in a single analysis. The mass accuracies measured for the precursor and product ions by the Orbitrap were <2 ppm in external mass calibration mode. Successful detection and characterization of GSH conjugates of acetaminophen, tienilic acid, clozapine, ticlopidine and mifepristone validated this method. In each case, the detected GSH conjugates were within the top five hits by isotope search. This method also has a broader detection capability since it is independent of the collision induced dissociation behavior of the GSH conjugates. Furthermore, this method is amenable to a broad class of reactive intermediate trapping agents as exemplified by the simultaneous detection and structural elucidation of the cyano-N-methylene iminium ion conjugates of verapamil and its O-desmethyl metabolites, which we report for the first time. In addition to the chemically tagged reactive intermediates, this method also provides information on stable metabolites from the full scan accurate MS data. PMID- 18383208 TI - Chromosomal imbalances, loss of heterozygosity, and immunohistochemical expression of TP53, RB1, and PTEN in intraductal cancer, intraepithelial neoplasia, and invasive adenocarcinoma of the prostate. AB - Recent studies have shown that intraductal prostate carcinoma (IDC-P) should be considered as a separate lesion distinct from prostatic intraepithelial neoplasia (PIN). The purpose of the present study was to analyze the genetic relationship between benign prostatic tissue, PIN, invasive cancer, IDC-P, and extracapsular tumor tissue to get further information about the role of IDC-P in the development of prostate cancer. One hundred five radical prostatectomy specimens were investigated immunohistochemically, 77 cases were analyzed by PCR for LOH of the tumor suppressor genes TP53 and RB1, and 11 cases of IDC-P and 10 cases of PIN were investigated using comparative genomic hybridization (CGH). At CGH analysis, IDC-P showed several chromosomal imbalances in contrast to PIN, where no changes were found. We could demonstrate a significant increase of LOH for TP53 or RB1 from benign tissue to PIN. LOH of both TP53 and RB1 were frequently found in IDC-P (52%), followed by extracapsular tumor tissue (44%), invasive cancer (24%), PIN (19%), and benign prostatic tissue (17%). Increased immunohistochemical expression was found in invasive cancer for TP53, RB1, and for PTEN. Decreased expression could be demonstrated in extracapsular tumor tissue and in IDC-P. Our results indicate that IDC-P in general follows the genetic pathway from normal epithelium over PIN lesion. IDC-P represents a separate prostatic lesion and should be graded as a poorly differentiated carcinoma. PMID- 18383209 TI - High-resolution analysis of genetic alterations in small bowel carcinoid tumors reveals areas of recurrent amplification and loss. AB - Carcinoid tumors of the small intestine are characterized by an indolent clinical course, secretion of neuropeptides, and resistance to standard cytotoxic chemotherapy. To evaluate the molecular events underlying carcinoid tumorigenesis, we used high-resolution arrays of single nucleotide polymorphisms to study chromosomal gains and losses in 24 primary and metastatic small bowel carcinoid tumors derived from 18 patients. Regions of gain or loss comprising whole chromosomes or large chromosomal regions constituted the most common class of anomalies. Loss of all or most of chromosome 18 was the commonest finding, evident in 11 of the 18 cases. Heterozygosity was also lost on chromosome arms 9p and 16q. The amplitude of observed gains was modest in comparison to those reported in some other tumor types. One focal region of recurrent gain on 14q mapped to the locus of the gene encoding the antiapoptotic protein DAD1, and immunohistochemical staining confirmed DAD1 protein expression in tumor samples. This detailed study of an uncommon neoplasm provides a basis to investigate putative oncogenes and tumor suppressor genes in intestinal carcinoid tumors. PMID- 18383210 TI - Detection of a t(1;22)(q23;q12) translocation leading to an EWSR1-PBX1 fusion gene in a myoepithelioma. AB - Chromosome banding as well as molecular cytogenetic methods are of great help in the diagnosis of mesenchymal tumors. Myoepithelial neoplasms of soft tissue including myoepitheliomas, mixed tumors, and parachordomas are diagnoses that have been increasingly recognized the last few years. It is still debated which neoplasms should be included in these morphologically heterogeneous entities, and the boundaries between them are not clear-cut. The pathogenetic mechanisms behind myoepithelial tumors are unknown. Only five parachordomas and one mixed tumor have previously been karyotyped, and nothing is known about their molecular genetic characteristics. We present a mesenchymal tumor classified as a myoepithelioma that had a balanced translocation t(1;22)(q23;q12) as the sole karyotypic change. A novel EWSR1-PBX1 fusion gene consisting of exons 1-8 of the 5'-end of EWSR1 and exons 5-9 of the 3'-end of PBX1 was shown to result from the translocation. Both genes are known to be targeted also by other neoplasia specific translocations, PBX1 in acute lymphoblastic leukemia and EWSR1 in several solid tumors, most of which are malignant. Based on the structure of the novel fusion gene detected, its transforming mechanism is thought to be the same as for other fusion genes involving EWSR1 or PBX1. PMID- 18383212 TI - Combination of off-line preparation and continuous flow mass spectrometry: D/H analyses of inclusion waters. PMID- 18383211 TI - Cells expressing partially unfolded R789C/p.R989C type II procollagen mutant associated with spondyloepiphyseal dysplasia undergo apoptosis. AB - We investigated the effects of the presence of R75C (p.R275C), R519C (p.719C), R789C (p.R989C), and G853E (p.G1053E) type II collagen (COL2A1) mutants, associated with distinct forms of spondyloepiphyseal dysplasia (SED), on the biological processes occurring in chondrocytic cells harboring those mutants. Mutant-specific biological responses of cells were initiated by activating tetracycline (Tet)-dependent expression of type II collagen mutants. Employing microscopic and biochemical assays, we determined that cells expressing the thermolabile R789C (p.R989C) type II collagen mutant undergo apoptosis. In contrast, in cells expressing the thermostable R75C (p.R275C), R519C (p.719C), and G853E (p.G1053E) mutants, apoptotic markers were not apparent. We also demonstrated that the R789C (p.R989C) mutant formed atypical complexes with endoplasmic reticulum (ER)-resident chaperones, thereby indicating an "unfolded protein response" (UPR) of cells harboring this specific mutant. Apoptotic changes were also demonstrated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and cleaved caspase 3 assays in the growth plates of mice harboring the R992C (p.R1147C) substitution in type II collagen. Based on these results, we propose that the intracellular presence of structurally altered type II collagen mutants could activate an apoptotic response, thereby limiting cell survival. By analyzing the response of cells to the altered structure of collagen mutants, our study contributes to better understanding the molecular basis of the pathological changes seen in vivo at the tissue level. PMID- 18383213 TI - Trypsin entrapped in poly(diallyldimethylammonium chloride) silica sol-gel microreactor coupled to matrix-assisted laser desorption/ionization time-of flight mass spectrometry. AB - An enzyme-immobilized capillary microreactor for rapid protein digestion and proteomics analysis is reported. The inner surface of the fused-silica capillary was coated with poly(diallyldimethylammonium chloride) (PDDA)-entrapped silica sol-gel matrix, followed by assembly of trypsin onto the PDDA-modified surface via electrostatic adsorption. The immobilization parameters such as PDDA content in the sol-gel matrix, trypsin concentration and pH were investigated in detail. Protein samples including beta-casein, myoglobin and cytochrome c could be effectively digested and electrophoretically separated simultaneously in such a modified capillary. Just 2.26 ng (corresponding to 0.10-0.14 picomole) of sample was sufficient for on-line capillary electrophoresis peptide mapping. The efficiency of the digestion was further demonstrated by digestion of a human liver cytoplasm sample and 253 proteins were identified in one unique run. PMID- 18383214 TI - A novel workflow control system for Fourier transform ion cyclotron resonance mass spectrometry allows for unique on-the-fly data-dependent decisions. AB - In this paper a novel workflow-based data acquisition and control system for Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) is presented that facilitates a fast on-the-fly decision-making process for a wide variety of data-dependent experiments. Several new workflow implementations demonstrate the flexibility and benefit of this approach for rapid dynamic experimental design on a chromatographic timescale. The different sequence, evaluation, decision and monitoring modules are described using a selected set of examples. During a tandem liquid chromatography (LC)/FTICR-MS experiment the system is used to dynamically switch between various dissociation techniques such as electron capture dissociation (ECD) and sustained off-resonance irradiation (SORI) depending on the charge state of a tryptic peptide peak. The use of this workflow-based system for imaging FTICR-MS using a desorption electrospray ionization (DESI) source demonstrates the possibility of external control of the workflow by feedback from an imaging sample stage. PMID- 18383215 TI - Electron ionization and fast atom bombardment mass spectral study for differentiation of ligand of zinc(II) (-)-sparteine and (-)-alpha-isosparteine complexes. PMID- 18383216 TI - Probing genetic algorithms for feature selection in comprehensive metabolic profiling approach. AB - Six different clones of 1-year-old loblolly pine (Pinus taeda L.) seedlings grown under standardized conditions in a green house were used for sample preparation and further analysis. Three independent and complementary analytical techniques for metabolic profiling were applied in the present study: hydrophilic interaction chromatography (HILIC-LC/ESI-MS), reversed-phase liquid chromatography (RP-LC/ESI-MS), and gas chromatography all coupled to mass spectrometry (GC/TOF-MS). Unsupervised methods, such as principle component analysis (PCA) and clustering, and supervised methods, such as classification, were used for data mining. Genetic algorithms (GA), a multivariate approach, was probed for selection of the smallest subsets of potentially discriminative classifiers. From more than 2000 peaks found in total, small subsets were selected by GA as highly potential classifiers allowing discrimination among six investigated genotypes. Annotated GC/TOF-MS data allowed the generation of a small subset of identified metabolites. LC/ESI-MS data and small subsets require further annotation. The present study demonstrated that combination of comprehensive metabolic profiling and advanced data mining techniques provides a powerful metabolomic approach for biomarker discovery among small molecules. Utilizing GA for feature selection allowed the generation of small subsets of potent classifiers. PMID- 18383220 TI - Photoconduction in amorphous organic solids. AB - Herein, we focus on the principles of photoconduction in random semiconductors the key processes being optical generation of charge carriers and their subsequent transport. This is not an overview of the current work in this area, but rather a highlight of elementary processes, their involvement in modern devices and a summary of recent developments and achievements. Experimental results and models are discussed briefly to visualize the mechanism of optical charge generation in pure and doped organic solids. We show current limits of models based on the Onsager theory of charge generation. After the introduction of experimental techniques to characterize charge transport, the hopping concept for transport in organic semiconductors is outlined. The peculiarities of the transport of excitons and charges in disorderd organic semiconductors are highlighted. Finally, a short discussion of ultrafast transport and single chain transport completes the review. PMID- 18383217 TI - Characterization of C-glycosyl quinochalcones in Carthamus tinctorius L. by ultraperformance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry. AB - C-Glycosyl quinochalcones are unique components in Carthamus tinctorius L. The reported C-glycosyl quinochalcones have the same quinochalcone skeleton with a hydroxyl group at the 5'-position and a glucose linked to this position with a carbon-carbon bond. In this study, the standard hydroxysafflor yellow A and water extracted fraction of Carthamus tinctorius L. were analyzed by ultraperformance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOFMS) in both positive and negative ion modes. The fragmentation pathways of C-glycosyl quinochalcones were interpreted and validated by accurate mass measurement. Their fragmentation showed a special cleavage at the C-C bond except for the typical internal cleavage at the sugar moiety of other C-glycosyl flavonoids. In positive ion mode, cleavage of the 5'-glucose produced an [M+H 162](+) ion by a neutral loss, while cleavage of the 5'-glucose in negative ion mode led to an [M-H-163](-.) ion by radical cleavage. The cleavage from the carbonyl group produced fragment ions containing an A or a B ring. The fragment ions containing an A ring were common product ions of seven compounds in both ion modes, and fragment ions containing the B ring were used to judge the different substituent groups at the 3'-position. The fragmentation patterns of seven structurally related C-glycosyl quinochalcones were analyzed systematically and the formation of the fragment ions in two modes is explained in detail in this report. UPLC/Q-TOFMS is an effective tool for characterizing a complex sample, which gives higher resolution separation and generates accurate mass measurement of the product ions. PMID- 18383221 TI - Influence of iron and chelator on siderophore production in Frankia strains nodulating Hippophae salicifolia D. Don. AB - Effect of iron and chelator on the growth and siderophore production in the ten newly Frankia strains isolated from the root nodules of Hippophae salicifolia D. Don and the two reference strains were studied. Growth of the strains was greatly affected when grown in the iron and EDTA deprived conditions. All the strains were capable of producing both the hydroxamate and catecholate type siderophore that was detected using the Csaky and Arnow assays. Production of siderophore was enhanced in the EDTA replenish condition in contrast to the iron supplemented medium suggesting that EDTA reduces the availability of other free metals and hence creates the stress condition for which the secretion of siderophore is enhanced. A decrease in siderophore production was observed with an increase in iron concentration. Strains HsIi2 and HsIi10 were found to be producing more siderophore than the other strains. PMID- 18383222 TI - Screening of Aspergillus species for occurrence of lectins and their characterization. AB - Ten species of Aspergillus were screened for occurrence of lectins. Each of the species was investigated for the occurrence of extracellular, surface-bound and intracellular lectin activities. As many as four species namely, Aspergillus niger, Aspergillus versicolor, Aspergillus rugulosus and Aspergillus nidulans, were found to possess intracellular lectin activities, while none of the species showed extracellular or surface-bound lectin activities. Each of the lectin was characterized with respect to blood group and carbohydrate specificities. All the lectins were found to agglutinate human erythrocytes, irrespective of their blood group and pig erythrocytes. However, they did not show agglutination with sheep or goat erythrocytes. Of the various carbohydrates tested, all lectins were found to be specific for inulin, mucin, asialofetuin, N-acetyl galactosamine, melibiose, D-ribose, L-fucose, D-arabinose, D-sucrose and D-mannitol. The minimum inhibitory concentration of each of the specific sugars was also determined. The lectins were partially purified using ammonium sulfate precipitation technique. Each of the lectin was found to be precipitated at 40-50% saturation of ammonium sulfate, yielding about 80% of lectin activity. PMID- 18383223 TI - Genetic diversity of endophytic bacteria which could be find in the apoplastic sap of the medullary parenchym of the stem of healthy sugarcane plants. AB - The genetic diversity of 29 endophytic bacterial strains isolated from apoplastic sap of the medullary parenchym of the stem of healthy sugarcane plants grown in Cuba was analysed by Two Primers-Ramdom Amplified Polymorphic DNA fingerprinting (TP-RAPD) and 16S rRNA gene sequencing. The strains were distributed into 17 groups on the basis of their TP-RAPD patterns, and a representative strain from each group was subjected to 16S rRNA gene sequencing. Analysis of these sequences showed that the isolates belong to a wide variety of phylogenetic groups being closely related to species of genera Bacillus and Staphylococcus from Firmicutes, Microbacterium, Micrococcus and Kokuria from Actinobacteria, Rhizobium and Gluconacetobacter from alpha -Proteobacteria, Comamonas and Xanthomonas from beta Proteobacteria, and Acinetobacter and Pantoea from gamma-Proteobacteria. These results show the complexity of the bacterial populations present in inner tissues of sugarcane, and indicate the interest and relevance of the studies on microbial diversity to improve our knowledge on the plant endophytic bacterial communities. PMID- 18383224 TI - Interaction between carbon and nitrogen metabolism during akinete development in the cyanobacterium Anabaena torulosa. AB - Nutrient enrichment with a nitrogen (as nitrate) or carbon (as fructose) source to unaerated diazo and photoautorophic cultures of the cyanobacterium Anabaena torulosa induced early development of akinetes with high frequency. When cultures under any mode of nutrition were aerated, akinetes were not differentiated. Unaerated cultures with nitrate nitrogen or fructose exhibited higher respiratory rates and nitrogen assimilation compared to aerated cultures. This was evidenced by increased respiratory O2 uptake and high activities of pyruvate kinase, malate dehydrogenase (NAD+), nitrogenase and nitrate reductase signifying that akinete forming unaerated cultures exhibited high carbon dissimilation and nitrogen assimilation resulting in high nitrogenous build up in the cells. Aerated, non akinete cultures, on the other hand, were associated with low respiratory O2 uptake, low pyruvate kinase and malate dehydrogenase (NAD+) activities, suggesting that carbon dissimilation was not favoured either in presence of nitrate or fructose. Moreover, higher activity of NADP+ linked malate dehydrogenase and lower nitrate reductase activity in aerated cultures led to a high carbon and low nitrogen content of the cells resulting in high cellular C:N ratio. The results suggest that interaction between carbon and nitrogen metabolism regulates akinete development in A. torulosa. PMID- 18383225 TI - Chemotaxis of Pseudomonas sp. to caffeine and related methylxanthines. AB - Pseudomonas sp. isolated from soil of coffee plantation area has been shown to degrade higher concentrations of caffeine ( approximately 15 g l(-1)) by N demethylation at a rate higher than what has been reported for any strain so far. This strain exhibits positive chemotaxis towards caffeine (1,3,7 trimethylxanthine) in swarm plate assay and modified capillary assay in a dose dependant manner. Related methylxanthines and xanthine also act as chemoattractants for the strain with the highest relative chemotactic response (RCR) seen for xanthine. Chemotaxis in Pseudomonas sp. is possibly plasmid mediated as indicated by positive chemotaxis of plasmid transformed E. coli DH5alpha. The chemotactic abilities of Pseudomonas sp. combined with higher rates of degradation of caffeine can be used in the development of strategies for biodecaffeination of caffeine containing wastes. PMID- 18383226 TI - Removal of chromium (VI) through biosorption by the Pseudomonas spp. isolated from tannery effluent. AB - Heavy metal contamination of the rivers is a world wide environmental problem and its removal is a great challenge. Kanpur and Unnao two closely located districts of Uttar Pradesh India are known for their leather industries. The tanneries release their treated effluent in the near by water ways containing Cr metal that eventually merges with the river Ganges. Untreated tannery effluent contains 2.673 +/- 0.32 to 3.268 +/- 0.73 mg l(-1) Cr. Microbes were isolated, keeping the natural selection in the view, from the tannery effluent since microbes present in the effluent exposed to the various types of stresses and metal stress is one of them. Investigations include the exposure of higher concentrations of Cr(VI) 1.0 to 4.0 mg l(-1) to the bacteria (presumably the Pseudomonas spp.) predominant on the agar plate. The short termed study (72 h) of biosorption showed significant reduction of metal in the media especially in the higher concentrations with a value from 1.0 +/- 0.02, 2.0 +/- 0.01, 3.0 +/- 0, and 4.0 +/- 0.09 at zero h to 0.873 +/- 0.55, 1.840 +/- 1.31, 2.780 +/- 0.03 and 3.502 +/ 0.68 at 72 h respectively. The biosorption of metal show in the present study that the naturally occurring microbes have enough potential to mitigate the excessive contamination of their surroundings and can be used to reduce the metal concentrations in aqueous solutions in a specific time frame. PMID- 18383227 TI - A new spiroketal from Aspergillus terreus, an endophytic fungus in Opuntia ficusindica Mill. AB - A new spiroketal, named aspergiketal (1) was isolated from the culture broth of Aspergillus terreus, an endophytic fungus in the stems of the plant Opuntia ficusindica Mill., together with two known compounds, physcion (2) and asterric acid (3). Their structures were elucidated by spectroscopic methods including 2D NMR techniques. PMID- 18383229 TI - Antagonistic effects of Pseudomonas jessenii against Pythium aphanidermatum: morphological, ultrastructural and cytochemical aspects. AB - Pseudomonas jessenii isolate EC-S101, an antagonistic rhizobacterium, induces morphological abnormalities such as topical swelling and excessive lateral branching in phytopathogenic Peronosporomycetes Pythium aphanidermatum hyphae as a result of radial growth inhibition in a dual culture assay. Rhodamine phalloidin staining revealed that these abnormalities were associated with disorganization of actin cytoskeleton. Both the morphological forms of actin, filaments and plaques, were affected progressively. At early stage of interaction (in less affected hyphae), the filaments were either eliminated or disarrayed. At advance stage of interaction (in severely affected hyphae), even the plaques population was decreased or disappeared. The effects of P. jessenii on actin architecture of Py. aphanidermatum were comparable to latrunculin B, a known actin assembly inhibitor. In addition, at early stage of interaction, the quantities of nuclei, lipid bodies and mitochondria became higher than those in control. At advance stage of interaction, the quantities of these organelles were almost similar, higher and lower, respectively, compare to those in control. Scanning electron microscopy exhibited cell wall disruption and accumulation of extracellular material associated with severely affected hyphae. Ultrastructural observations of the affected hyphae displayed additional features of considerable thickening of cell wall, enlargement of vacuoles, sinking of redundant lipid bodies into the enlarged vacuoles and wall appositions. We conclude that in addition to interference in morphogenesis and growth of Py. aphanidermatum, P. jessenii suppresses the pathogen through sub-cellular disorganization, specifically the actin architecture. This is the first report on disruption of cytoskeleton in a eukaryotic phytopathogen by an antagonistic rhizobacterium. PMID- 18383230 TI - Mycoflora associated with Hyoscyamus muticus growing under an extremely arid desert environment (Aswan region, Egypt). AB - Hyoscyamus muticus L. (Egyptian henbane) is one of the desert medicinal plants of family Solanaceae. The plant produces pharmaceutically important compounds (tropane alkaloids) as secondary metabolites. In the present study, we describe mycoflora of H. muticus grown in four different locations in Egyptian southern desert (Aswan region): Aswan university campus, Wadi Allaqi down stream part, Aswan airport road, and Sahari city. Eighty-one species and two varieties belonging to 31 genera were isolated from soils surrounding H. muticus plants, the surface of the plants, and inside the plants as endophytic fungi. Aspergillus was the most common genus in all study areas. The highest number of genera and species of fungi were recorded in Aswan university campus followed by Aswan airport road. Fungal diversity analysis revealed that these two locations have higher fungal diversity compared to other two locations. A higher number of fungal species were isolated from rhizosphere soil and rhizoplane than from non rhizosphere and other plant organs. Endophytic fungi were isolated from all plant parts of H. muticus. Communications between H. muticus plants and fungi under desert conditions both in rhizosphere and inside the plant are deduced. PMID- 18383231 TI - Studies on the heterogeneity of the carotenogenic yeast Rhodotorula mucilaginosa from Patagonia, Argentina. AB - The yeast species Rhodotorula mucilaginosa is considered to be ubiquitous due to its world-wide distribution in terrestrial, freshwater and marine habitats, and to its ability to colonize a large variety of substrates. In this paper we assess the phenotypic and genetic variability of environmental isolates of R. mucilaginosa collected from natural and artificial environments in Patagonia, Argentina. A total of 97 strains were studied and sorted into three groups based on MSP-PCR fingerprinting results: A, which comprised 90% of the strains, including the type strain; and B and C which included 2 and 8% of the strains, respectively. The D1D2 sequencing did not differentiate any of the 3 groups, while ITS sequencing validated the existence of group C. This group was composed of Patagonian isolates of diverse origin. DNA-DNA reassociation studies confirmed the existence of significant genetic differences between group C and the type strain. In this study, which is the first on the intraspecific variability of a large set of R. mucilaginosa isolates, a considerable phenotypic and genetic heterogeneity was observed, however such differences were not enough to refute co specificity. The study of Patagonian isolates allowed the detection of a genetically distinct group of R. mucilaginosa strains. PMID- 18383232 TI - Influence of farnesol on the morphogenesis of Aspergillus niger. AB - Farnesol was the first quorum-sensing regulator to be found in eukaryotic cells. In Candida albicans, a dimorphic fungal human pathogen, farnesol blocks the yeast to-filamentous growth transition. Here we show that in Aspergillus niger farnesol acts as an inhibitor of conidiation: Colonies grown on media containing farnesol were unable to develop conidia. Although farnesol treated A. niger cultures exhibited a colony morphology resembling the "fluffy" phenotype of A. nidulans, which is caused by a hyperactive G-protein/cAMP pathway, the intracellular level of cAMP in A. niger mycelia grown in presence of farnesol is greatly diminished. Furthermore, whereas inhibiting adenylyl cyclase led to a farnesol-like effect, the addition of external cAMP inhibited conidiation without causing a "fluffy" phenotype. This suggests that the mechanisms regulating conidiation in A. niger and A. nidulans are different. PMID- 18383234 TI - Leptin, adiponectin, resistin, and ghrelin--implications for inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) is characterized by anorexia, malnutrition, altered body composition, and development of mesenteric white adipose tissue (WAT) hypertrophy. Increasing evidence suggests that adipokines synthesized either in WAT or in immune cells, are involved in these manifestations of IBD. Among adipokines leptin, adiponectin and resistin hold a fundamental role while the role of ghrelin in inflammation is not well established. Preliminary studies have shown overexpression of leptin, adiponectin, and resistin in mesenteric WAT of patients with Crohn's disease (CD) and significant alterations of circulating serum levels of these adipokines in IBD. It has also been demonstrated that intestinal inflammation causes an increase in endogenous ghrelin production. In animal models of intestinal inflammation, existing data suggest that leptin, adiponectin, and resistin are pivotal mediators of inflammation. Interesting therapeutic interventions based on these data have been suggested. A specific role for hypertrophic WAT has also been implicated in CD. Further efforts with experimental and clinical studies are needed to better understand the role of adipokines in IBD. PMID- 18383235 TI - Prebiotics, synbiotics and inflammatory bowel disease. AB - The normal colonic microflora is intimately involved in the aetiology of inflammatory bowel diseases (IBD) such as ulcerative colitis (UC) and Crohn's disease (CD). These conditions are often refractile to conventional treatments involving the employment of anti-inflammatory and immunosuppressant drugs, and this has led to a search for alternative therapies based on the use of probiotics, prebiotics and synbiotics. The majority of investigations in this area have been done with probiotics, and while there is increasing interest in the abilities of prebiotics and synbiotics to control the symptoms of IBD, very few randomised controlled trials have been reported. Although the results have been variable, human and animal studies have demonstrated that in many circumstances, these functional foods can alter the composition of the colonic microbiota, reduce inflammatory processes in the gut mucosa, and have the potential to induce disease remission. More work is needed to understand the effects of prebiotics and synbiotics on microbial communities in the gut, and their interactions with the host's immune system. PMID- 18383236 TI - Organelle tracking in a living cell with microsecond time resolution and nanometer spatial precision. AB - The study of cellular processes such as organelle transport often demands particle tracking with microsecond time-resolution and nanometer spatial precision, posing significant challenges to existing tracking methods. Here, we have developed a novel strategy for two-dimensional tracking of gold nanoparticles (GNPs) with 25 mus time resolution and approximately 1.5 nm spatial precision, by using a quadrant photodiode to record the positions of GNPs in an objective-type dark-field microscope. In combination with a feedback loop, this technique records long, high time-resolution and spatial precision trajectories of endocytosed GNPs transported by the molecular motors kinesin and dynein in a living cell. In the full range of organelle velocities (0-8 microm s(-1)), we clearly resolve the individual 8 nm steps of cargoes carried by kinesin, and the 8, 12, 16, 20, and 24 nm steps of those carried by dynein. These experiments yield new information about molecular motor stepping in living cells. PMID- 18383237 TI - Thin-film properties of DNA and RNA bases: a combined experimental and theoretical study. AB - The structures of the DNA and RNA bases cytosine, uracil, and thymine in thin films with a nominal film thickness of about 20 nm are studied by using X-ray photoemission spectroscopy (XPS) and Fourier-transform infrared spectroscopy. The molecules are evaporated in situ from powder on a gold foil. The experimental results indicate that cytosine is composed of two energetically close tautomeric forms, whereas uracil and thymine exist in only one tautomeric form. Additionally, quantum chemical calculations are performed to complement the experimental results. The relative energies of the tautomeric forms of cytosine, uracil, and thymine are calculated using Hartree-Fock (HF), density functional theory (DFT), and post-HF methods. Furthermore, the assignment of the XPS spectra is supported by using simple model considerations employing Koopmans ionization energies and Mulliken net atomic charges. PMID- 18383238 TI - Ordered mesoporous thin films of rutile TiO2 nanocrystals mixed with amorphous Ta2O5. AB - Ordered mesoporous thin films of composites of rutile TiO2 nanocrystals with amorphous Ta2O5 are fabricated by evaporation-induced self-assembly followed by subsequent heat treatment beyond 780 degrees C. Incorporation of selected amounts of Ta2O5 (20 mol %) in the mesoporous TiO2 film, together with the unique mesoporous structure itself, increased the onset of crystallization temperature which is high enough to ensure the crystallization of amorphous titania to rutile. The ordered mesoporous structure benefits from a block-copolymer template, which stabilizes the mesostructure of the amorphous mixed oxides before crystallization. The surface and in-depth composition analysis by X-ray photoelectron spectroscopy suggests a homogeneous intermixing of the two oxides in the thin film. A detailed X-ray absorption fine structure measurement on the composite film containing 20 mol % Ta2O5 and heated to 800 degrees C confirms the amorphous nature of the Ta2O5 phase. Photocatalytic activity evaluation suggests that the rutile nanocrystals in the synthesized ordered mesoporous thin film possess good ability to assist the photodegradation of rhodamine B in water under illumination by UV light. PMID- 18383239 TI - Probing innovative microfabricated substrates for their reproducible SERS activity. AB - New types of microfabricated surface-enhanced Raman spectroscopy (SERS) active substrates produced by electron beam lithography and ion beam etching are introduced. In order to achieve large enhancement factors by using the lightning rod effect, we prepare arrays consisting of sharp-edged nanostructures instead of the commonly used dots. Two experimental methods are used for fabrication: a one stage process, leading to gold nanostar arrays and a two-stage process, leading to gold nanodiamond arrays. Our preparation process guarantees high reproducibility. The substrates contain a number of arrays for practical applications, each 200x200 microm2 in size. To test the SERS activity of these nanostar and nanodiamond arrays, a monolayer of the dye crystal violet is used. Enhancement factors are estimated to be at least 130 for the nanodiamond and 310 for the nanostar arrays. PMID- 18383240 TI - Reinvestigation of a cyclic dipeptide N-prenyltransferase reveals rearrangement of N-1 prenylated indole derivatives. PMID- 18383241 TI - A novel genetic selection system for improved enantioselectivity of Bacillus subtilis lipase A. AB - In directed evolution experiments, success often depends on the efficacy of screening or selection methods. Genetic selections have proven to be extremely valuable for evolving enzymes with improved catalytic activity, improved stability, or with altered substrate specificity. In contrast, enantioselectivity is a difficult parameter to select for. In this study, we present a successful strategy that not only selects for catalytic activity, but for the first time also for enantioselectivity, as demonstrated by the selection of Bacillus subtilis lipase A variants with inverted and improved enantioselectivity. A lipase mutant library in an aspartate auxotroph Escherichia coli was plated on minimal medium that was supplemented with the aspartate ester of the desired enantiomer (S)-(+)-1,2-O-isopropylidene-sn-glycerol. To inhibit growth of less enantioselective variants, a covalently binding phosphonate ester of the opposite (R)-(-)-1,2-O-isopropylidene-sn-glycerol enantiomer was added as well. After three selection rounds in which the selection pressure was increased by raising the phosphonate ester concentration, a mutant was selected with an improved enantioselectivity increased from an ee of -29.6 % (conversion 23.4 %) to an ee of +73.1 % (conversion 28.9 %) towards the (S)-(+)-enantiomer. Interestingly, its amino acid sequence showed that the acid of the catalytic triad had migrated to a position further along the loop that connects beta7 and alphaE; this shows that the position of the catalytic acid is not necessarily conserved in this lipase. PMID- 18383242 TI - Semipreparative chiral supercritical fluid chromatography in the fractionation of lansoprazole and two related antiulcer drugs enantiomers. AB - The semipreparative chiral separation of lansoprazole and two related compounds (pantoprazole and rabeprazole) using supercritical fluid chromatography (SFC) is presented in this work. Different loads were evaluated in order to obtain high enantiomeric purities and production rates. The volumes injected were 1, 2 and 4 mL. The concentrations of the racemic mixtures were 3 and 6 g/L for lansoprazole and 1.5 g/L for pantoprazole and rabeprazole. In all the cases, the recoveries, for a purity higher than 99.9%, were better for the second eluted enantiomer than for the first one. This fact conditioned the production rate of the first eluted enantiomer that, considering a fixed purity, was always lower than that obtained for the other one. In the case of lansoprazole it was possible to obtain 0.025 and 0.090 mg/min of the first and second eluted enantiomer, respectively, with an enantiomeric purity of 99.9%. For rabeprazole enantiomers 0.037 and 0.062 mg/min, and in the case of pantoprazole the results were better (0.062 and 0.122 mg/min) due to the higher resolution. PMID- 18383243 TI - Supercritical fluid extraction of cynaropicrin and 20-hydroxyecdysone from Leuzea carthamoides DC. AB - Leuzea carthamoides is an adaptogenic plant containing biologically active compounds as ecdysteroids and guaianolide-type sesquiterpene lactones, conventionally extracted from the plant with ethanol. It may be a potential source of the mentioned natural compounds. Ethanol-modified near-critical CO(2) was used as selective solvent with the aim to increase the level of 20 hydroxyecdysone in the extract from L. carthamoides roots and to remove selectively cynaropicrin, a sesquiterpene lactone of bitter taste, from the leaves. The extraction conditions were varied (pressure 20-28 MPa, temperature 40 60 degrees C, ethanol concentration in the solvent 0-7.1%) and the extraction yield and extract composition were compared with the results of ethanolic extraction. The supercritical fluid extraction (SFE) from finely powdered plant was controlled by phase equilibrium. Cynaropicrin was quantitatively removed from the leaves where 89% of 20-hydroxyecdysone was retained. The extraction yield of 20-hydroxyecdysone from roots with ethanol-modified CO(2 )was lower by 30% than with ethanol but its concentration in the extract was higher by 67%. PMID- 18383244 TI - Extraction of carotenoids and chlorophyll from microalgae with supercritical carbon dioxide and ethanol as cosolvent. AB - The extraction of carotenoids and chlorophylls using carbon dioxide modified with ethanol as a cosolvent is an alternative to solvent extraction because it provides a high-speed extraction process. In the study described here, carotenoid and chlorophyll extraction with supercritical CO(2 )+ ethanol was explored using freeze-dried powders of three microalgae (Nannochloropsis gaditana, Synechococcus sp. and Dunaliella salina) as the raw materials. The operation conditions were as follows: pressures of 200, 300, 400 and 500 bar, temperatures of 40, 50 and 60 degrees C. Analysis of the extracts was performed by measuring the absorbance and by using empirical correlations. The results demonstrate that it is necessary to work at a temperature of 50-60 degrees C and a pressure range of 300-500 bar, depending on the type of microalgae, in order to obtain the highest yield of pigments. The best carotenoid/chlorophyll ratios were obtained by using supercritical fluid extraction + cosolvent instead of using conventional extraction. The higher selectivity of the former process should facilitate the separation and purification of the two extracted pigments. PMID- 18383245 TI - An improved interface for universal acoustic flame detection in modified supercritical fluid chromatography. AB - A novel method of interfacing the acoustic flame detector (AFD) with modified supercritical fluid chromatography (SFC) is presented. By applying resistive heating directly to the burner region between the restrictor outlet and the acoustic flame, infrequent severe noise, baseline drifting, and peak deformations that can occasionally be observed with the AFD are eliminated. For example, by increasing the interface temperature only a few hundred degrees Celsius, such sporadic noise in the detector can be reduced nearly ten-fold resulting in smooth stable operation of the AFD. Further, for various levels of methanol modified supercritical carbon dioxide mobile phase examined, the interface was observed to reduce detector noise in each to a common minimal range near 10-25 Hz when an appropriate temperature was achieved. The method is simply assembled, inexpensive to construct, and robust in its daily operation. Overall, the heated interface developed and presented facilitates reliable AFD operation in modified SFC, and supports further exploration and implementation of this sensor as an alternative universal detector in separations requiring an organic cosolvent in the mobile phase. PMID- 18383246 TI - Effects of ozone exposure in rat lungs investigated with hyperpolarized 3 He MRI. AB - PURPOSE: To investigate the effects of subchronic ozone exposure on rat lung ventilation using hyperpolarized (HP) (3)He MRI. MATERIALS AND METHODS: A total of 24 Sprague-Dawley rats, distributed in one control group and four groups exposed to 0.5 ppm ozone concentration for two days or six days, either continuously (22 hours/day) or alternatingly (12 hours/day). A three-step MRI protocol was designed and applied to each animal, including: 1) (3)He gas distribution images acquired at inspiratory capacity, 2) measurements of intrapulmonary (3)He diffusion coefficients, and 3) dynamic ventilation acquisitions performed during lung filling with (3)He. RESULTS: No differentiation between animals exposed to ozone and control animals was observed from the ventilation images obtained at inspiratory capacity. The (3)He diffusion coefficients were not statistically different from one group to another. Ventilation defects, appearing as delayed lung filling regions and heterogeneous lung filling, were observed in the dynamic lung ventilation image series. The percentage of animals with ventilation defects in the control, two-day, and six day exposed groups were equal to 20%, 43% and 75%, respectively. In the subgroup of the animals exposed six days for 12 hours per day, the percentage of animals exhibiting ventilation defects was equal to 85%. CONCLUSION: Heterogeneous obstructive patterns in an experimental animal model of subchronic ozone exposure were observed using HP (3)He MRI. PMID- 18383247 TI - Reduction of truncation artifacts in rapid 3D articular cartilage imaging. AB - PURPOSE: To reduce Gibbs ringing artifact in three-dimensional (3D) articular knee cartilage imaging with linear prediction (LP). MATERIALS AND METHODS: A reconstruction method using LP in 3D was applied to truncated data sets of six healthy knees. The technique first linearizes the data before applying the prediction algorithm. Three radiologists blindly reviewed and ranked images of the full, truncated, and predicted data sets. Statistical analysis of the radiologists' reviews was performed for image quality, clinical acceptability of the images, and equivalence with the gold standard. RESULTS: LP applied to 3D knee cartilage imaging allows for 40% decreased scan time while providing image quality with statistical equivalence to a full data set. CONCLUSION: 3D spoiled gradient echo imaging (SPGR) knee cartilage imaging requires significant scan time. This 40% reduction in scan time will allow such scans to be more feasible without sacrificing clinical acceptability. PMID- 18383248 TI - Physiological noise in MR images: an indicator of the tissue response to ischemia? AB - PURPOSE: To determine whether measuring signal intensity (SI) fluctuations in MRI time series data from acute stroke patients would identify ischemic tissue. MATERIALS AND METHODS: Prebolus perfusion-weighted MRI data from 32 acute ischemic stroke patients (N = 32) was analyzed as a time series. Ischemic and normal tissue regions were outlined and compared. RESULTS: The magnitude of the measured SI fluctuations was significantly lower in ischemic regions relative to normal tissue. Spatial differences in these fluctuations occurred in a manner that was different than other perfusion-based metrics. CONCLUSION: Prior studies have shown that SI fluctuations in MRI time series data correspond to the presence of physiological "noise," which includes vasomotion, an autoregulatory phenomenon that affects the tissue response to ischemia. In this study, SI fluctuations were found to decrease in ischemia, consistent with the notion that small vessels will remain open (fluctuations in vessel diameter will decrease) when there is a challenge to flow. Spatial variation in SI fluctuations appeared to be different from spatial variation seen on other perfusion-based metrics, suggesting that a separate contrast mechanism is responsible, one that might be of diagnostic and prognostic value in acute stroke in which the ability of tissue to withstand ischemia is currently not well visualized. PMID- 18383249 TI - Contrast-enhanced MRI of murine sponge model for progressive angiogenesis assessed with gadoteridol (ProHance) and gadocoletic acid trisodium salt (B22956/1). AB - PURPOSE: To investigate the potential value of MRI for noninvasive assessment of angiogenesis in a murine model exploiting the properties of two contrast agents, gadoteridol (ProHance) and gadocoletic acid trisodium salt (B22956/1). MATERIALS AND METHODS: Biocompatible sponges were implanted in both mice flanks. Stimulated sponges contained human recombinant basic fibroblast growth factor (bFGF) as the angiogenic agent; control sponges contained vehicle. Angiogenesis was evaluated by MRI after injection of extravascular (ProHance) or blood-pool (B22956/1) contrast agents at different times after sponge implantation. Sponges signal intensity enhancement was calculated both as the relative enhancement and the rate of relative enhancement. Results from MRI were validated by classic biochemical (hemoglobin level and protein content) and morphological (histology) assays. RESULTS: The intrinsic different properties of ProHance and B22956/1 in wash-in and wash-out kinetics were useful to detect progressive vascularization and the establishment of a functional vascular network in the implants. Moreover, MRI allowed the appreciation of differences in neovessel colonization between bFGF-treated sponges and controls. Hemoglobin level, protein content, and histology confirmed the sponge vascularization and MRI results. CONCLUSION: Contrast-enhanced MRI is a reliable tool to study vascular characteristics in animal models of angiogenesis. The different kinetic properties of contrast agents can provide evidence of different functional neovascularization aspects and levels. PMID- 18383250 TI - Source of low-frequency fluctuations in functional MRI signal. AB - PURPOSE: To investigate the source of native low-frequency fluctuations (LFF) in functional MRI (fMRI) signal. MATERIALS AND METHODS: Phase analysis was performed on tissue-segmented fMRI data acquired at systematically varying sampling rates. RESULTS: LFF in fMRI signal were both native and aliased in origin. Scanner instability did not contribute to native or aliased LFF. Aliased LFF arose from cardiorespiratory processes and head motion. Native LFF did not arise from cardiorespiratory processes, but did so, at least in part, from head motion. Motion correction reduced native LFF, but did not eliminate them. The residual native LFF in motion-corrected fMRI data showed a systematic phase difference among different tissue structures. The native LFF in fMRI signals of cerebral blood vessels and CSF were synchronous, and preceded those of gray and white matter, indicating that the vascular fluctuations lead the metabolic fluctuations. CONCLUSION: The primary physiologic source of native LFF in fMRI signal is vasomotion. PMID- 18383251 TI - Numerical and experimental study of a novel phase contrast magnetic resonance (PC MR) imaging technique: sparse interleaved referencing PC-MR imaging. AB - PURPOSE: To use numerical simulation and experimental approaches to introduce a novel phase contrast magnetic resonance (PC-MR) data processing technique termed Sparse Interleaved Referencing PC-MR, with potential to improve accuracy, temporal resolution, and signal-to-noise ratio (SNR) of PC-MR data. MATERIALS AND METHODS: Computational fluid dynamics data were generated for a two-chamber orifice flow model simulating valvular regurgitation. The numerical results were validated and used to simulate conventional and Sparse Interleaved Referencing PC MR data acquisitions. Common data sets were processed using conventional and Sparse Interleaved Referencing approaches and quantitative errors in velocity time waveforms were measured and compared. In vitro phantom jet flow data and in vivo ascending aorta data were acquired and used to simulate Sparse Interleaved Referencing PC-MR. RESULTS: The Sparse Interleaved Referencing PC-MR data showed significantly better representation of the velocity-time waveform in three areas: (i) lower root-mean-square errors (9.0 +/- 1.0% versus 24.0 +/- 0.2%; P < 0.005), (ii) simulation of conventionally processed data showed a pattern of peak velocity overestimation, which was experimentally demonstrated in in vitro data, whereas overestimation of peak velocity was dramatically attenuated using Sparse Interleaved Referencing (2.8 +/- 0.4% versus 16.9 +/- 6.4%, P < 0.005), and (iii) compared with the conventional scan, an average of 119.4 +/- 26.6% (P < 0.005) SNR was realized in in vitro and in vivo Sparse Interleaved Referencing PC-MR data. CONCLUSION: Simulation and in vitro/in vivo results show that Sparse Interleaved Referencing PC-MR processed data in pulsatile and jet flow showed higher accuracy, better peak velocity representation, and improved SNR compared with the data processed using the conventional PC-MR method. PMID- 18383252 TI - Local re-inversion coronary MR angiography: arterial spin-labeling without the need for subtraction. AB - PURPOSE: To implement a double-inversion bright-blood coronary MR angiography sequence using a cylindrical re-inversion prepulse for selective visualization of the coronary arteries. MATERIALS AND METHODS: Local re-inversion bright-blood magnetization preparation was implemented using a nonselective inversion followed by a cylindrical aortic re-inversion prepulse. After an inversion delay that allows for in-flow of the labeled blood-pool into the coronary arteries, three dimensional radial steady-state free-precession (SSFP) imaging (repetition/echo time, 7.2/3.6 ms; flip angle, 120 degrees, 16 profiles per RR interval; field of view, 360 mm; matrix, 512, twelve 3-mm slices) is performed. Coronary MR angiography was performed in three healthy volunteers and in one patient on a commercial 1.5 Tesla whole-body MR System. RESULTS: In all subjects, coronary arteries were selectively visualized with positive contrast. In addition, a middle-grade stenosis of the proximal right coronary artery was seen in one patient. CONCLUSION: A novel T1 contrast-enhancement strategy is presented for selective visualization of the coronary arteries without extrinsic contrast medium application. In comparison to former arterial spin-labeling schemes, the proposed magnetization preparation obviates the need for a second data set and subtraction. PMID- 18383254 TI - Alteration of proton diffusivity associated with passive muscle extension and contraction. AB - PURPOSE: To determine whether passive muscle extension and contraction affect the proton diffusivity of the muscle. MATERIALS AND METHODS: Five male subjects were examined. The fractional anisotropy (FA), and primary (lambda(1)), secondary (lambda(2)), and tertiary eigenvalues (lambda(3)) of the right tibialis anterior and medial gastrocnemius muscles were compared between conditions of passive plantar flexion and passive dorsiflexion of the ankle joint. RESULTS: In the tibialis anterior, FA, and lambda(1) at dorsiflexion decreased significantly (P < 0.01 and P < 0.01, respectively) compared to those at plantar flexion, but lambda(3) at dorsiflexion increased significantly (P = 0.02). In the gastrocnemius, FA and lambda(1) at dorsiflexion increased significantly (P < 0.01 and P < 0.01, respectively) compared to those at plantar flexion, but lambda(3) at dorsiflexion decreased significantly (P < 0.01). The lambda(2) value showed no significant change in either the tibialis anterior or medial gastrocnemius. CONCLUSION: The results indicate that passive muscle extension and contraction associated with passive joint movement would affect the proton diffusivity of the muscle. This alteration of proton diffusivity is probably associated with microscopic structural changes of the muscle. PMID- 18383253 TI - Comparison between 2D and 3D high-resolution black-blood techniques for carotid artery wall imaging in clinically significant atherosclerosis. AB - PURPOSE: To compare two- (2D) and three-dimensional (3D) black-blood imaging methods for morphological measurements of the carotid artery wall and atherosclerotic plaque. MATERIALS AND METHODS: A total of 18 subjects with 50% to 79% carotid stenosis were scanned with 2D (2-mm slice thickness) and 3D (1-mm/0.5 mm actual/interpolated slice thickness) T1-weighted fast spin-echo (FSE) black blood imaging sequences with double inversion-recovery (DIR) blood suppression. Morphological measurements (lumen area, wall area, vessel area, mean wall thickness, and maximal wall thickness), signal-to-noise ratio (SNR) in the wall and lumen, and wall-lumen contrast-to-noise ratio (CNR) were compared between 2D and 3D images. The effect of improved slice resolution in 3D imaging was evaluated for visualization of small plaque components. RESULTS: Lumen SNR (P = 0.16), wall SNR (P = 0.65), and CNR (P = 0.94) were comparable between 2D/3D. There was no difference in average lumen area (P = 0.16), average wall area (P = 0.99), average vessel area (P = 0.0.58), mean wall thickness (P = 0.09), and maximum wall thickness (P = 0.06) between 2D/3D. Distributions of small plaque components such as calcification were better characterized by the 3D acquisition. There was a higher sensitivity to motion artifacts with 3D imaging, resulting in three examinations with low image quality. CONCLUSION: 2D and 3D protocols provided comparable morphometric measurements of the carotid artery. The major advantage of 3D imaging is improved small plaque component visualization, while the 2D technique provides higher reliability for image quality. PMID- 18383255 TI - Human MRI above the FDA 8 T guideline: can we conclude that it is safe? PMID- 18383257 TI - Hyperpolarized 129 Xe MRI of the mouse lung at a low xenon concentration using a continuous flow-type hyperpolarizing system. AB - PURPOSE: To apply a continuous flow-type hyperpolarizing (CF-HP) system to lung imaging and investigate the feasibility of hyperpolarized (129)Xe MRI at a low xenon concentration. MATERIALS AND METHODS: Under two conditions where a 3% or 70% xenon gas mixture was constantly supplied, gas- and dissolved-phase (129)Xe images and diffusion-weighted (129)Xe-gas images were obtained from the mouse lung. Signal-to-noise ratio (SNR) of the (129)Xe images and the apparent diffusion coefficient (ADC) of xenon were compared between the two gas mixtures. RESULTS: The SNR of gas- and dissolved-phase images were 28.9 +/- 5.2 and 12.0 +/ 2.0, respectively, using the 70% xenon gas mixture, while they were 22.9 +/- 4.8 and 6.8 +/- 0.6, using the 3% mixture. The ADC of xenon using the 3% xenon gas mixture was approximately 1.5 times higher than that using the 70% one. These results indicated that the high ADC increases the apparent replenishment rate of gas-phase magnetization, thus resulting in a reduction of the SNR loss induced by diluting xenon with quenching gases. CONCLUSION: The CF-HP system is useful for lung imaging at an extremely low concentration of xenon, which enables one to fully restrain an anesthetic effect of xenon and to reduce consumption of xenon in a measurement. PMID- 18383256 TI - Two-dimensional 1H MR spectroscopy of the brain in human immunodeficiency virus (HIV)-infected children. AB - PURPOSE: To measure cerebral metabolites in brains of human immunodeficiency virus (HIV)-infected patients using two-dimensional (2D) proton ((1)H) magnetic resonance spectroscopy (MRS), which enables more sensitive detection of metabolites at lower concentrations and delineation of the components of the different choline (Ch) groups in the frequency domain when compared to one dimensional (1D) (1)H-MRS. MATERIALS AND METHODS: We examined metabolite/creatine (Cr) and metabolite/Ch ratios in the left frontal brain of 10 HIV-infected (mean age 13.7 +/- 4.7 years) and 11 control (mean age 15.3 +/- 4.6 years) adolescents and children using 2D localized chemical shift correlated spectroscopy (L-COSY). The integrated volume under each 2D metabolite peak was calculated with reference to the diagonal creatine methyl peak (Cr_d) or the diagonal choline trimethylamine peak (Ch_d). RESULTS: In the HIV-infected patients, myoinositol (mI)/Cr_d (P = 0.009) and mI/Ch_d (P = 0.006) were elevated. The ratios of the following metabolites were also significantly elevated (P < 0.05): mI-Ch/Cr_d, gamma-aminobutyrate (GABA)/ Cr_d, GABA/Ch_d, threonine-lactate (Thr-Lac)/Cr_d, Thr-Lac/Ch_d, and N-acetyl aspartate (NAA)/Cr_d. CONCLUSION: We have demonstrated for the first time the feasibility of 2D-MRS in HIV-infected children and adolescents to assess cerebral metabolites and found elevated mI and elevated GABA, in the left frontal brain of clinically stable HIV-infected patients. A larger study population is needed to confirm these pilot GABA findings. PMID- 18383258 TI - Simultaneous imaging of myocardial motion and chamber blood flow with SPAMM n' EGGS (Spatial Modulation of Magnetization With Encoded Gradients for Gauging Speed). AB - PURPOSE: To provide simultaneous measurements of one-dimensional (1-D) myocardial displacement and 1-D chamber blood flow in a single breath-held acquisition using an MR imaging technique, SPAMM n' EGGS (Spatial Modulation of Magnetization With Encoded Gradients for Gauging Speed). MATERIALS AND METHODS: Velocity encoding bipolar gradients sensitive to chamber blood flow were played out before the readout gradient in a 1-1 SPAMM-tagged MR imaging pulse sequence. For any given motion-flow encoded direction, the acquired image sequence was later postprocessed to separate the tag motion and blood flow terms. Experiments were performed on seven normal volunteers, and two pigs with moderate ischemic mitral regurgitation. Left-ventricular motion and trans-valvular flow obtained using the SPAMM n' EGGS pulse sequence was compared against measurements obtained using standard tagging and phase-contrast pulse sequences, respectively. RESULTS: Results in normal volunteers and diseased pigs demonstrate multiphase correlated measurements of myocardial motion and chamber blood flow using SPAMM n' EGGS. A close correspondence in these measurements to conventional tagging and phase contrast sequences is confirmed. CONCLUSION: We have demonstrated that simultaneous acquisition of myocardial motion and chamber blood flow is possible within a single breath-hold. The data obtained using the SPAMM n' EGGS pulse sequence may be useful in the planning and evaluation of mitral-valve repair procedures. PMID- 18383259 TI - Quantification of MRI measured myocardial perfusion reserve in healthy humans: a comparison with positron emission tomography. AB - PURPOSE: To validate a noninvasive quantitative MRI technique, the K(i) perfusion method, for myocardial perfusion in humans using (13)N-ammonia PET as a reference method. MATERIALS AND METHODS: Ten healthy males (64 +/- 8 years) were examined with combined PET and MRI perfusion imaging at rest and during stress induced by dipyridamole in order to determine the myocardial perfusion reserve. Myocardial and blood time concentration curves obtained by Gd-DTPA-enhanced MRI and (13)N ammonia PET were fitted by a two-compartment perfusion model. RESULTS: Mean perfusion values (+/-SD) derived from the MRI method at rest and at hyperemia were 80 +/- 20 and 183 +/- 56 mL/min/100 g, respectively. The same data for PET were 71 +/- 16 and 203 +/- 67 mL/min/100 g. A linear relationship was observed between MRI and PET-derived myocardial perfusion reserve for regional and global data. Linear regression for the global absolute perfusion reserve gave a correlation coefficient of 0.96 (P < 0.004, y=0.83x-6.9). A good agreement between the two methods to determine low or high perfusion reserves was found. CONCLUSION: Our data provide validation of the perfusion marker K(i) derived by the MRI method as a quantitative marker for myocardial perfusion in healthy humans. PMID- 18383261 TI - Evaluating local hepatocellular carcinoma recurrence post-transcatheter arterial chemoembolization: is diffusion-weighted MRI reliable as an indicator? AB - PURPOSE: To evaluate the detectability of local hepatocellular carcinoma (HCC) recurrence after transcatheter arterial chemoembolization (TACE) by diffusion weighted MR imaging in correlation with those of gadolinium-enhanced MR imaging. MATERIALS AND METHODS: Respiratory-triggered diffusion-weighted MR images (b factor, 500 s/mm(2); number of averaging, six were obtained in 25 patients with 39 HCCs. Two independent radiologists evaluated diffusion-weighted MR images, gadolinium-enhanced MR images after TACE, and assigned confidence levels for postoperative HCC recurrence. Apparent diffusion coefficients (ADCs) in HCCs were also measured. Sensitivities and specificities were compared using an extension of the McNemar test. Observer performance was also determined by ROC curve analysis. RESULTS: Local recurrences in 14 HCCs and complete tumor necrosis in 25 HCCs after TACE were determined. Sensitivity for the detection of local HCC recurrence was higher on gadolinium-enhanced MR imaging (82%) than on diffusion weighted MR imaging (60.7%) for the two readers in combination and separately (P < 0.05). Specificities were comparably high for both sequences. Az values were higher for gadolinium-enhanced MR images (0.92) than for diffusion-weighted MR images (0.74) for readers in combination and separately (P < 0.05). Mean ADC values showed an increase after TACE (P < 0.001). CONCLUSION: Diffusion-weighted MR imaging was not found to be a reliable predictor of local HCC recurrence after TACE as compared with gadolinium-enhanced MR imaging. PMID- 18383260 TI - Estrogen receptor and breast MR imaging features: a correlation study. AB - PURPOSE: To compare the MRI features between estrogen receptor (ER) positive and negative breast cancers. MATERIALS AND METHODS: Breast MRI of 90 consecutive patients confirmed with invasive ductal carcinoma (IDC), 51 ER positive and 39 ER negative, were analyzed. The tumor morphology and dynamic contrast-enhanced (DCE) kinetics were evaluated based on the American College of Radiology (ACR) Breast Imaging Reporting and Data System (BI-RADS) MRI lexicon and compared. Enlarged axillary lymph nodes on MRI and choline (Cho) detection using MR spectroscopy (MRS) were also analyzed and compared. For patients receiving axillary node dissection the pathological nodal status was also compared. RESULTS: ER negative breast cancer had bigger tumors compared to ER positive cancer (3.6 +/- 2.0 cm vs. 1.8 +/- 1.3 cm, P < 0.00005). ER negative cancer was more likely to exhibit nonmass type enhancements compared to ER positive cancer (P < 0.005). Enlarged axillary lymph nodes were more frequently identified on MRI in ER negative compared to ER positive patients (P < 0.05). After excluding patients undergoing neoadjuvant chemotherapy, auxiliary lymph node status did not show significant difference between ER positive and ER negative cancer on MRI and pathology. ER negative cancer was more likely to show the malignant type enhancement kinetics (P = 0.15), rim enhancement (P = 0.15), and Cho detection on MRS (P = 0.23) compared to ER positive cancer, but it did not reach a level of statistical significance. CONCLUSION: ER negative breast cancer was more aggressive, with larger tumor size, more non-mass-type enhancement lesions, and a higher percentage showing enlarged axillary nodes on MRI. These features might be related to its poorer cellular differentiation and/or a higher angiogenesis. PMID- 18383262 TI - Capillary permeability and extracellular volume fraction in uterine cervical cancer as patient outcome predictors: measurements by using dynamic MRI spin lattice relaxometry. AB - PURPOSE: To improve the outcome prediction of uterine cervical carcinoma by measuring the vascular permeability (k(ep)) and the extracellular volume fraction (v(e)) of the tumor from Dynamic T(1)- IRM Relaxometry. MATERIALS AND METHODS: Twenty-six patients with proven cervical carcinoma were divided into good outcome and poor outcome groups. Classic tumor prognostic factors, the longest diameter L and the volume V of the tumor, were measured from morphologic MR images. The tumor parameters k(ep) and v(e) were determined from the relaxometry time-curve acquired during the contrast uptake after a bolus intravenous injection of an extracellular contrast agent. RESULTS: All "small" tumors (L<35 mm or V<11 cm(3)) were good outcome with 100% sensitivity but a rather low specificity (36% and 43% for L and V, respectively). With regard to the physiopathological parameter k(ep), "large" tumors (L >or= 35 mm) can also be classified as good outcome on the condition that k(ep) >or= 2.2 min(-1) with 100% sensitivity and 89% specificity. Regarding the extracellular volume fraction (v(e)), no significant difference was observed between the two groups. CONCLUSION: Measurement of the tumor vascular permeability might be useful to predict prognostic, to evaluate the treatment efficacy, and to adapt a proper therapy schedule. PMID- 18383263 TI - In vivo ultra-high-field magnetic resonance imaging of trabecular bone microarchitecture at 7 T. AB - PURPOSE: To investigate the feasibility of 7 T magnetic resonance imaging (MRI) to visualize and quantify trabecular bone structure in vivo by comparison with 3T MRI and in vivo three-dimensional (3D) high-resolution peripheral quantitative computed tomography (HR-pQCT). MATERIALS AND METHODS: The distal tibiae of 10 healthy volunteers were imaged. Therefore, fully balanced steady state free precession (bSSFP) and spin-echo (bSSSE) pulse sequences were implemented and optimized for 7 T. Structural bone parameters, such as apparent bone-volume over total-volume fraction (app.BV/TV), apparent trabecular plate separation (app.TbSp), apparent trabecular plate thickness (app.TbTh), and apparent trabecular plate number (app.TbN), were derived. RESULTS: All structural trabecular bone parameters correlated well (r > 0.6) between 7T and 3T, and between 7 T and HR-pQCT (r > 0.69), with the exception of app.TbTh, which correlated modestly (r = 0.41) between field strengths and very low with HR-pQCT (r < 0.16). Regarding absolute values, app.TbN varied only 4% between field strengths, and only 0.6% between 7 T and HR-pQCT. App.TbSp correlated best between 7 T and HR-pQCT (r = 0.89). Using bSSSE, significant smaller trabecular thickness and significant higher trabecular number were found compared to bSSFP. CONCLUSION: We concluded that imaging and quantification of the trabecular bone architecture at 7 T is feasible and preferably done using bSSSE. There exists great potential for ultra-high-field (UHF) MRI applied to trabecular bone measurements. PMID- 18383266 TI - Minimally invasive precision brain access using prospective stereotaxy and a trajectory guide. AB - PURPOSE: To evaluate the capabilities of MR-guided "prospective stereotaxy" methods for accessing brain structures for biopsy or electrode implantation. MATERIALS AND METHODS: MR-guided biopsy and deep brain stimulator (DBS) electrode implantations were performed with a trajectory guide and real-time MR guidance. Imaging methods were used to plan the selected path through the brain, appropriately orient the trajectory guide, and monitor the device insertion to assure technical success and screen for hemorrhage. Assessments of technical success rate, targeting accuracy, and complications associated with this technique were performed. RESULTS: A total of 187 biopsy procedures were performed with guidance via prospective stereotaxy methods. All brain biopsies were diagnostic and two patients sustained superficial wound infections that were treated successfully with antibiotics. One patient died postoperatively of a myocardial infarction despite preoperative medical clearance. A total of 42 DBS electrode insertions were performed in patients with Parkinson's disease or dystonia. The difference between planned and actual electrode position averaged 1.2 mm +/- 0.7 mm on the first pass and only a single brain penetration was required in 90% of electrode insertions. Complications included a single asymptomatic hemorrhage and two early infections, with the latter addressed by an adjustment to sterile practice. CONCLUSION: Prospective stereotaxy, in combination with a trajectory guide, has been proven capable of efficiently and accurately targeting structures throughout the brain. PMID- 18383265 TI - Relationship between choline and apparent diffusion coefficient in patients with gliomas. AB - PURPOSE: To examine the relationship between apparent diffusion coefficients (ADC) from diffusion weighted imaging (DWI) and choline levels from proton magnetic resonance spectroscopic imaging (MRSI) in newly diagnosed Grade II and IV gliomas within distinct anatomic regions. MATERIALS AND METHODS: A total of 37 patients with Grade II and 28 patients with Grade IV glioma were scanned on a 1.5T system with 3D MRSI and DWI. Region level analysis included Spearman rank correlation between median normalized ADC and choline for each patient per grade within each distinct abnormal anatomical region. Voxel level analysis calculated a Spearman rank correlation per region, per patient. RESULTS: Grade II lesions showed no evidence of a correlation between normalized ADC and choline using either the region or voxel level analysis. Region level analysis of Grade IV lesions did not appear to correlate in the contrast enhancement or necrotic core, but did suggest a significant negative correlation in the more heterogeneous nonenhancing and combined regions. CONCLUSION: There appears to be differences in the relationship between ADC and choline levels in Grade II and Grade IV gliomas. Correlation within these regions in Grade IV lesions was strongest when all regions were included, suggesting heterogeneity may be driving the relationship. PMID- 18383267 TI - High temporal resolution functional MRI using parallel echo volumar imaging. AB - PURPOSE: To combine parallel imaging with 3D single-shot acquisition (echo volumar imaging, EVI) in order to acquire high temporal resolution volumar functional MRI (fMRI) data. MATERIALS AND METHODS: An improved EVI sequence was associated with parallel acquisition and field of view reduction in order to acquire a large brain volume in 200 msec. Temporal stability and functional sensitivity were increased through optimization of all imaging parameters and Tikhonov regularization of parallel reconstruction. Two human volunteers were scanned with parallel EVI in a 1.5T whole-body MR system, while submitted to a slow event-related auditory paradigm. RESULTS: Thanks to parallel acquisition, the EVI volumes display a low level of geometric distortions and signal losses. After removal of low-frequency drifts and physiological artifacts, activations were detected in the temporal lobes of both volunteers and voxelwise hemodynamic response functions (HRF) could be computed. On these HRF different habituation behaviors in response to sentence repetition could be identified. CONCLUSION: This work demonstrates the feasibility of high temporal resolution 3D fMRI with parallel EVI. Combined with advanced estimation tools, this acquisition method should prove useful to measure neural activity timing differences or study the nonlinearities and nonstationarities of the BOLD response. PMID- 18383268 TI - Dynamic contrast-enhanced quantitative perfusion measurement of the brain using T1-weighted MRI at 3T. AB - PURPOSE: To develop a method for the measurement of brain perfusion based on dynamic contrast-enhanced T(1)-weighted MR imaging. MATERIALS AND METHODS: Dynamic imaging of the first pass of a bolus of a paramagnetic contrast agent was performed using a 3T whole-body magnet and a T(1)-weighted fast field echo sequence. The input function was obtained from the internal carotid artery. An initial T(1) measurement was performed in order to convert the MR signal to concentration of the contrast agent. Pixelwise and region of interest (ROI)-based calculation of cerebral perfusion (CBF) was performed using Tikhonov's procedure of deconvolution. Seven patients with acute optic neuritis and two patients with acute stroke were investigated. RESULTS: The mean perfusion value for ROIs in gray matter was 62 mL/100g/min and 21 mL/100g/min in white matter in patients with acute optic neuritis. The perfusion inside the infarct core was 9 mL/100g/min in one of the stroke patients. The other stroke patient had postischemic hyperperfusion and CBF was 140 mL/100g/min. CONCLUSION: Absolute values of brain perfusion can be obtained using dynamic contrast-enhanced MRI. These values correspond to expected values from established PET methods. Furthermore, at 3T pixelwise calculation can be performed, allowing construction of CBF maps. PMID- 18383269 TI - Non-bioengineered silk gland fibroin protein: characterization and evaluation of matrices for potential tissue engineering applications. AB - The possibility of using wild non-mulberry silk protein as a biopolymer remains unexplored compared to domesticated mulberry silk protein. One of the main reasons for this was for not having any suitable method of extraction of silk protein fibroin from cocoons and silk glands. In this study non-bioengineered non mulberry silk gland fibroin protein from tropical tasar silkworm Antheraea mylitta, is regenerated and characterized using 1% (w/v) sodium dodecyl sulfate (SDS). The new technique is important and unique because it uses a mild surfactant for fibroin dissolution and is advantageous over other previous reported techniques using chaotropic salts. Fabricated fibroin films are smooth as confirmed by atomic force microscopy. Circular dichroism spectrometry along with Fourier transformed infrared spectroscopy and X-ray diffraction reveal random coil/alpha-helix conformations in regenerated fibroin which transform to beta-sheets, resulting in crystalline structure and protein insolubility through ethanol treatment. Differential scanning calorimetry shows an increase in glass transition (Tg) temperature and enhanced degradation temperature on alcohol treatment. Enhanced cell attachment and viability of AH927 feline fibroblasts were observed on fibroin matrices. Higher mechanical strength along with controllable water stability of regenerated gland fibroin films make non-mulberry Indian tropical tasar silk gland fibroin protein a promising biomaterial for tissue engineering applications. PMID- 18383270 TI - Impact of clarification strategy on chromatographic separations: Pre-processing of cell homogenates. AB - This research focused on how the extent and type of primary solid-liquid separation can affect the performance of guard filtration and chromatography, in this instance hydrophobic interaction chromatography. The system used in the study was yeast (Saccharomyces cerevisiae) with the target molecule being an intracellular protein; alcohol dehydrogenase (ADH). As expected, loading more poorly clarified suspensions (both centrates and primary filtrates) required proportionally larger guard filtration areas. In addition for feed suspensions prepared by centrifugation, increased clarification led to greater column capacity. However, where filtration was used to achieve similar clarification considerably lower column capacity was achieved. These results were attributed to centrifugation leading to the aggregation of lipids and their subsequent removal in this form before application to the column. Clarification by filtration leaves such lipids in their original "soluble" state and hence they are not removed. The importance of the need to examine such interactive effects in bioprocess studies is discussed. This observation was confirmed with further analytical work into the nature of the aggregated material formed in the supernatant under centrifugation conditions. This material was only soluble in an organic solvent, and identified as phophatidylcholine and ergosterol as among the components removed by centrifugation and guard filtration as opposed to filtration and guard filtration. PMID- 18383271 TI - Electronic volume of CD34 positive cells from peripheral blood apheresis samples. AB - BACKGROUND: The authors have used a flow analyzer to measure electronic cellular volume of peripheral blood hematopoietic stem/progenitor cells obtained by granulocyte-colony stimulating factor (G-CSF) mobilization and apheresis (HPC-A) of patients with hematological malignancies. METHODS: Fifty three apheresis samples stained with CD45-fluorescein isothiocyanate (FITC) and CD34-R phycoerythrin (PE)-labeled antibodies after erythrocyte lysis with BD FACS Lysing Solution were analyzed for electronic cell volume and two-color FITC and PE fluorescence. RESULTS: Lymphocytes, monocytes and granulocytes in the HPC-A samples had a mean electronic volume of 414, 797, and 670 microm(3), respectively corresponding to cell diameter of 9.25, 11.5, and 10.85 microm. In 53 HPC-A samples analyzed, the mean electronic volume of the CD34 positive mononuclear cells was 407 microm(3) while the CD45 positive cells had mean volume of 453 microm(3). CONCLUSIONS: CD34 positive stem/progenitor cells have a smaller volume and diameter than CD45 positive mononuclear cells in HPC-A samples. In the present study the CD34 stem/progenitor cells had a considerably larger diameter than that of stem cells previously reported in the literature. With the availability of electronic cell volume as a parameter in flow cytometric analysis, further studies can be carried out to correlate stem cell volume with specific phenotypic marker expression. PMID- 18383272 TI - Reproductive anatomy of the female Amazonian manatee Trichechus inunguis Natterer, 1883 (Mammalia: Sirenia). AB - The Amazonian manatee (Trichechus inunguis) is uniparous and has a slow reproduction cycle due to a long gestation period and long interval between births. Even though protected by law, hunting remains one of the main causes hindering the natural population growth of this species in the wild. The histology and reproductive anatomy provide information on the history and reproductive status of the female and offer a tool for the conservation of the species. The present study describes the anatomy of the female reproductive tract in T. inunguis. It is based on materials from three reproductive tracts fixed in 10% buffered formalin. The ovaries, uterine tubes, uterus, vagina, and external genitalia are described. The hymen presents two tiny openings separated by a segment that, upon rupturing during the first copulation, should make up a single vaginal opening. A still intact hymen and the absence of placental scars in the uterus were found in one specimen. Additionally, the presence of a hemorrhagic body and Graafian follicles on the right ovary were observed, as well as whitish scars and among them, possible corpora albicantia. These findings suggest that T. inunguis undergoes infertile estrus cycles before its first gestation. Macroscopically, counting of the whitish scars is hindered by the small diameter of these structures. It is not possible to differentiate between the scars resulting from ruptured (corpora albicantia) and nonruptured follicles (regressed corpora atretica). The presence of whitish scars on both ovaries of the same specimen suggests their bilateral function in T. inunguis. PMID- 18383273 TI - Liquid and gas chromatography coupled to isotope ratio mass spectrometry for the determination of 13C-valine isotopic ratios in complex biological samples. AB - On-line gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) is commonly used to measure isotopic ratios at natural abundance as well as for tracer studies in nutritional and medical research. However, high-precision (13)C isotopic enrichment can also be measured by liquid chromatography-isotope ratio mass spectrometry (LC-IRMS). Indeed, LC-IRMS can be used, as shown by the new method reported here, to obtain a baseline separation and to measure (13)C isotopic enrichment of underivatised amino acids (Asp, Thr-Ser, Glu, Pro, Gly, Ala, Cys and Val). In case of Val, at natural abundance, the SD(delta(13)C) reported with this method was found to be below 1 per thousand . Another key feature of the new LC-IRMS method reported in this paper is the comparison of the LC-IRMS approach with the conventional GC-C-IRMS determination. To perform this comparative study, isotopic enrichments were measured from underivatised Val and its N(O, S)-ethoxycarbonyl ethyl ester derivative. Between 0.0 and 1.0 molar percent excess (MPE) (delta(13)C= -12.3 to 150.8 per thousand), the calculated root-mean-square (rms) of SD was 0.38 and 0.46 per thousand and the calculated rms of accuracy was 0.023 and 0.005 MPE, respectively, for GC-C-IRMS and LC-IRMS. Both systems measured accurately low isotopic enrichments (0.002 atom percent excess (APE)) with an SD (APE) of 0.0004. To correlate the relative (delta(13)C) and absolute (atom%, APE and MPE) isotopic enrichment of Val measured by the GC-C IRMS and LC-IRMS devices, mathematical equations showing the slope and intercept of the curves were established and validated with experimental data between 0.0 to 2.3 MPE. Finally, both GC-C-IRMS and LC-IRMS instruments were also used to assess isotopic enrichment of protein-bound (13)C-Val in tibial epiphysis in a tracer study performed in rats. Isotopic enrichments measured by LC-IRMS and GC-C IRMS were not statistically different (p>0.05). The results of this work indicate that the LC-IRMS was successful for high-precision (13)C isotopic measurements in tracer studies giving (13)C isotopic enrichment similar to the GC-C-IRMS but without the step of GC derivatisation. Therefore, for clinical studies requiring high-precision isotopic measurement, the LC-IRMS is the method of choice to measure the isotopic ratio. PMID- 18383274 TI - pCREB is involved in neural induction of mouse embryonic stem cells by RA. AB - Mouse embryonic stem (ES) cells can be induced by various chemicals to differentiate into a variety of cell types in vitro. In our study, retinoic acid (RA), one of the most important inducers, used at a concentration of 5 microM, was found to induce the differentiation of ES cells into neural progenitor cells (NPCs). During embryoid body (EB) differentiation, the level of active cyclic AMP response element-binding protein (CREB) was relatively high when 5 microM RA treatment was performed. Inhibition of CREB activity committed EBs to becoming other germ layers, whereas increased expression of CREB enhanced NPC differentiation. Moreover, RA increased the expression of active CREB by enhancing the activity of JNK. Our research suggests that CREB plays a role in RA induced NPC differentiation by increasing the expression of active JNK. PMID- 18383275 TI - Multiresidue method for the analysis of more than 140 pesticide residues in fruits and vegetables by gas chromatography coupled to triple quadrupole mass spectrometry. AB - A new multiresidue method has been developed and validated for the determination of more than 140 pesticide residues in cucumber and orange by gas chromatography coupled to triple quadrupole mass spectrometry (GC-QqQ-MS/MS) in a single run of 25.50 min. The triple quadrupole (QqQ) analyzer simultaneously operated in the selected reaction monitoring (SRM) and selected ion monitoring (SIM) modes, acquiring two or three transitions per compound. Samples were extracted by the application of a single-phase extraction of 10 g of sample with acetonitrile containing 1% of acetic acid, followed by a liquid-liquid partition formed by the addition of 4 g of MgSO(4) and 1 g of NaOAc. A dispersive solid-phase extraction (D-SPE) with primary secondary amine (PSA) was applied to clean up the extracts. A final concentration step was included in order to increase sensitivity in the instrumental analysis. The method was properly validated in each matrix in a wide dynamic range (10-400 microg kg(-1)): this work relies on a new quantification strategy by the use of two calibration curves to increase the dynamic range, which permitted reduction of sample dilutions and increase in sample throughput. Recovery was studied at three concentration levels (11.5, 50.0, and 150.0 microg kg(-1)), yielding values in the range 70-110% with precision values, expressed as relative standard deviation (RSD), lower than 20 and 25% for the intraday and interday precision, respectively. Limits of quantification (LOQs) were established at 10 microg kg(-1), the lowest maximum residue level (MRL) value set by the European Union in vegetables. The method was successfully applied to the analysis of pesticide residues in real samples from the southeastern Spain. PMID- 18383276 TI - Comparative qualitative and quantitative assessment of biomineralization of tooth development in man and zebrafish (Danio rerio). AB - It was the aim of this study to investigate the distribution of Ca, P, and C in predentin, mineralizing dentin, and mature dentin of human tooth buds and compare these results with those of zebrafish (Danio rerio) teeth using energy dispersive X-ray analysis (EDX) element analysis. The mandible of a 16-week-old human fetus containing 6 mineralizing tooth buds and three complete heads of zebrafish were fixed in formaldehyde and embedded in Technovit 9100. Serial sections of 80 microm thickness were cut in frontal-dorsal direction, and from these sections, polarized light micrographs were taken. The sections with tooth buds were then investigated with scanning electron microscopy, and EDX element analysis was performed with a Philips XL 30 FEG scanning microscope and an EDAX energy dispersive X-ray system using spot measurements, EDX line-scans and element mapping. Quantitative measurements were made in predentin, mineralizing dentin adjacent to predentin, and mature dentin. The Ca and P content increased rapidly from outer predentin toward mineralizing dentin in human tooth buds and in zebrafish teeth. The Ca/P ratio was different for predentin and dentin areas, reflecting different calcium phosphate compositions in predentin and fully mineralized dentin. Because of the similarities between human tooth buds and zebrafish teeth, it can be concluded that the zebrafish tooth development may be an excellent model for studying biomineralization processes during odontogenesis. PMID- 18383277 TI - Endocranial occipito-temporal anatomy of SD-1219 from the Neandertal El Sidron Site (Asturias, Spain). AB - We addressed the brain drainage system as inferred by the endocranial morphology of the occipito-temporal region of the El Sidron Neandertal specimen SD-1219. Morphological details of the endocranial surface and its anatomical implications were analyzed for the reconstruction of the dural sinus drainage pattern and its comparison with Neandertals and other hominids. The specimen SD-1219 shows a pattern in which the superior sagittal sinus goes into the right transverse sinus. Comparative analyses with a large sample of fossil hominids reveal a pattern of the SD-1219 fossil that is typical for Neandertals. The analysis of the proportions of the occipital lobes prints within the occipital fossae reveals that the left occipital pole projects toward the right. This possibly indicates brain asymmetry (petalia) in this Neandertal individual, similar to that observed in some modern human brains. Conversely, no such asymmetry was observed in the cerebellar fossae. A particular feature of this fossil is the presence of two crests, located at the middle of the left cerebellar fossa that can be related to either an imprinting of a cerebellar fissure or some bone response to mechanical influence on internal bone surface morphology during cerebellar development. Specific aspects of the paleoneurology of Neandertals are discussed. Further quantitative studies on the endocranial morphology of the occipito-temporal and mastoid region will shed light on the paleoneurological significance of this important anatomical region for the understanding of human evolution. PMID- 18383278 TI - Tuberculosis induced changes to the osseous cranial base and its potential effect on hearing. AB - Our prior work suggested that petro-occipital fissure (POF) ossification may be altered in clinicopathologies of the cranial base such as hearing loss (Balboni et al., 2005). Here we demonstrate an accelerated and statistically significant ossification of the POF and cochlear aqueduct (CA) in a historical population of patients diagnosed with tuberculosis (TB). While a number of studies have sought to reduce the importance of the POF/CA to hearing, given its anatomical location, evolutionary conservation across mammals and the mounting data linking morphological changes of the POF/CA to the temporal onset of hearing loss and tinnitus, it is becoming difficult to maintain that its function is not related to inner ear homeostasis. PMID- 18383279 TI - Origin and endpoint of the olfactory nerve fibers: as described by Santiago Ramon y Cajal. AB - In the late Nineteenth Century, Santiago Ramon y Cajal was able to reproduce an exceptional illustration of the Olfactory Nerve pathway and its myriad of cells, by using the Golgi Method. Dr. Cajal focused intense study on the histology of the nervous system and published a treatise on the olfactory nerve fibers and the distinct peripheral origin and central nervous system endpoint of this unique pathway. The original title of this work is "Origen y terminacion de las fibras nerviosas olfatorias" published in 1890. As the original publication is in Spanish, here we provide an English translation allowing present-day English speakers to read these writings. Cajal followed the trajectory of the olfactory nerve fibers as they transitioned between the peripheral and central nervous system and was able to assert that these fibers were not continuous from the olfactory bulb to the bipolar cells that relinquish into the olfactory epithelium, but that the olfactory system was made up of various cell types each having distinct morphologies and functions. This may very well be the first definitive description of the olfactory receptor neurons and the first illustrations of the continuity of these cells throughout the olfactory pathway. These meticulous histological preparations were created by first using Camillo Golgi's potassium dichromate and silver nitrate impregnation method known as "reazione nera" or "black reaction," where nerve cells, nerve fibers, and neuroglia could be visualized. This study exhibits the structural and functional organization of the mammalian fila olfactoria as it was investigated in centuries past. PMID- 18383280 TI - Modification of embolic-PVA particles with MR contrast agents. AB - We report the synthesis and characterization of polyvinyl alcohol (PVA) embolic particles modified with a clinically approved magnetic resonance (MR) contrast agent. PVA particles are used during transcatheter arterial embolization (TAE) procedures and this minimally invasive technique is a widely employed treatment for inoperable tumors. The PVA particles are injected into tumor vessels and prevent blood flow which results in tumor attenuation. An accurate assessment of the endpoint of embolization is critical to successful TAE procedures. Recent reports suggest that 20% of endpoint determination of TAE procedures by angiographic techniques are erroneous. Real time, in vivo imaging of the embolic particles would overcome this limitation. The contrast-modified PVA particles described here show an 80% decrease in T(1) relaxation times compared to unmodified particles. Images of particles in capillary tubes of similar size to catheters used in TAE procedures are clearly visible by MRI. PMID- 18383281 TI - Magnitude least squares optimization for parallel radio frequency excitation design demonstrated at 7 Tesla with eight channels. AB - Spatially tailored radio frequency (RF) excitations accelerated with parallel transmit systems provide the opportunity to create shaped volume excitations or mitigate inhomogeneous B(1) excitation profiles with clinically relevant pulse lengths. While such excitations are often designed as a least-squares optimized approximation to a target magnitude and phase profile, adherence to the target phase profile is usually not important as long as the excitation phase is slowly varying compared with the voxel dimension. In this work, we demonstrate a method for a magnitude least squares optimization of the target magnetization profile for multichannel parallel excitation to improve the magnitude profile and reduce the RF power at the cost of a less uniform phase profile. The method enables the designer to trade off the allowed spatial phase variation for the improvement in magnitude profile and reduction in RF power. We validate the method with simulation studies and demonstrate its performance in fourfold accelerated two dimensional spiral excitations, as well as for uniform in-plane slice selective parallel excitations using an eight-channel transmit array on a 7T human MRI scanner. The experimental results are in good agreement with the simulations, which show significant improvement in the magnitude profile and reductions in the required RF power while still maintaining negligible intravoxel phase variation. PMID- 18383282 TI - Iterative GRAPPA (iGRAPPA) for improved parallel imaging reconstruction. AB - In this work an iterative reconstruction method based on generalized autocalibrating partially parallel acquisitions (GRAPPA) reconstruction is introduced. In the new method the reconstructed lines are used to reestimate and refine the weights from all the acquired data by applying the GRAPPA procedure iteratively with regularization. Both phantom and in vivo MRI experiments demonstrated that, compared to GRAPPA, the iterative approach reduces parallel imaging artifacts and permits high-quality image reconstruction with a relatively small number of calibration lines and slight changes of GRAPPA weights. PMID- 18383283 TI - Spinal cord blood flow measurement by arterial spin labeling. AB - The assessment of spinal cord (SC) hemodynamics, and especially SC blood flow (SCBF), plays a key role in the pathophysiological description and understanding of many SC diseases such as ischemia, or spinal cord injury. SCBF has been previously measured in animals with invasive techniques such as autoradiography or labeled microspheres; no MR technique, however, has been proposed so far. The possibility of quantitatively measuring SCBF in mice using MRI was investigated using a presaturated FAIR (flow-sensitive alternating inversion recovery) arterial spin labeling (ASL) technique. SCBF measurements were performed at the cervical level of the mouse as well as on the brain so as to use cerebral blood flow (CBF) values as internal references. With a spatial resolution of 133 x 133 microm(2) for the SCBF maps, absolute regional perfusion values could be measured within the different structures of the SC (gray matter, white matter, and cerebrospinal fluid area). Similar perfusion values were found in SC gray matter (330+/-90 mL/100g/min) and in brain (295+/-22 mL/100g/min for thalamus). This result, in agreement with SCBF/CBF measurements performed with non-MR techniques, opens new perspectives for noninvasive longitudinal and in vivo animal studies. Application to human experiments may also be possible. PMID- 18383284 TI - Slice-selective images of free radicals in mice with modulated field gradient electron paramagnetic resonance (EPR) imaging. AB - Continuous wave (CW) electron paramagnetic resonance (EPR) imaging can be used to obtain slice-selective images of free radicals without measuring three dimensional (3D) projection data. A method that incorporated a modulated magnetic field gradient (MFG) was combined with polar field gradients to select a slice in the subject noninvasively. The slice-selective in vivo EPR imaging of triarylmethyl radicals in the heads of live mice is reported. 3D surface-rendered images were successfully obtained from slice-selective images. In the experiment in mice, a slice thickness of 1.8 mm was achieved. PMID- 18383285 TI - Imaging brain vasculature with BOLD microscopy: MR detection limits determined by in vivo two-photon microscopy. AB - Rat brain vasculature was imaged at 9.4T with blood oxygenation level-dependent (BOLD) microscopy. Data were acquired without exogenous contrast agent in <35 min using 3D gradient-echo imaging with 78-microm isotropic resolution. Detailed vascular patterns including intracortical veins and some branches were observed in simple magnitude-contrast data acquired at an experimentally optimized echo time. The venous origin of the dark patterns was confirmed by oxygenation dependent studies, and when the systemic arterial oxygen saturation level was <80% BOLD microscopy revealed additional intracortical vessels presumed to be of arterial origin. Quantification shows a decrease of intracortical venous density with depth. The full width at half-minimum intensity was 90-190 microm for most intracortical venous vessels identifiable by BOLD venography. Since actual diameters are not directly quantifiable by BOLD, we also measured diameter dependent intracortical venous density in vivo by two-photon excitation fluorescent microscopy. Density comparisons between the two modalities, along with computer simulations, show that venous vessels as small as approximately 16 30 microm diameter are detectable with 9.4T BOLD microscopy under our experimental conditions. PMID- 18383286 TI - Robust MRI brain tissue parameter estimation by multistage outlier rejection. AB - This article addresses the problem of the tissue type parameter estimation in brain MRI in the presence of partial volume effects. Automatic MRI brain tissue classification is hampered by partial volume effects that are caused by the finite resolution of the acquisition process. Due to this effect intensity distributions in brain MRI cannot be well modeled by a simple mixture of Gaussians and therefore more complex models have been developed. Unfortunately, these models do not seem to be robust enough for clinical conditions, as the quality of the tissue classification decreases rapidly with the image quality. Also, the application of these methods for pathological images with unmodeled intensities (e.g. MS plaques, tumors, etc.) remains uncertain. In the present work a new robust method for brain tissue characterization is presented, treating the partial volume affected voxels as outliers of the pure tissue distributions. The proposed method estimates the tissue characteristics from a reduced set of intensities belonging to a particular pure tissue class. This reduced set is selected by using a trimming procedure based on local gradient information and distributional data. This feature makes the method highly tolerant of a large amount of unexpected intensities without degrading its performance. The proposed method has been evaluated using both synthetic and real MR data and compared with state-of-the-art methods showing the best results in the comparative. PMID- 18383288 TI - Additive angle method for fast large-tip-angle RF pulse design in parallel excitation. AB - Current methods for parallel excitation RF pulse design are based on the small tip-angle approximation, which provides a computationally efficient means of pulse calculation. In general, pulses designed with those methods are inaccurate when scaled to produce large-tip angles, and methods for large-tip-angle pulse design are more computationally demanding. This paper introduces a fast iterative method for large-tip-angle parallel pulse design that is formulated as a small number of Bloch equation simulations and fast small-tip-angle pulse designs, the results of which add to produce large-tip-angle pulses. Simulations and a phantom experiment demonstrate that the method is effective in designing multidimensional large-tip-angle pulses of high excitation accuracy, compared to pulses designed with small-tip-angle methods. PMID- 18383287 TI - Differentiation between benign and malignant breast lesions detected by bilateral dynamic contrast-enhanced MRI: a sensitivity and specificity study. AB - The purpose of this study was to apply an empirical mathematical model (EMM) to kinetic data acquired under a clinical protocol to determine if the sensitivity and specificity can be improved compared with qualitative BI-RADS descriptors of kinetics. 3D DCE-MRI data from 100 patients with 34 benign and 79 malignant lesions were selected for review under an Institutional Review Board (IRB) approved protocol. The sensitivity and specificity of the delayed phase classification were 91% and 18%, respectively. The EMM was able to accurately fit these curves. There was a statistically significant difference between benign and malignant lesions for several model parameters: the uptake rate, initial slope, signal enhancement ratio, and curvature at the peak enhancement (at most P=0.04). These results demonstrated that EMM analysis provided at least the diagnostic accuracy of the kinetic classifiers described in the BI-RADS lexicon, and offered a few key advantages. It can be used to standardize data from institutions with different dynamic protocols and can provide a more objective classification with continuous variables so that thresholds can be set to achieve desired sensitivity and specificity. This suggests that the EMM may be useful for analysis of routine clinical data. PMID- 18383289 TI - Pittfalls of MRI measurement of white matter perfusion based on arterial spin labeling. AB - Although arterial spin labeling (ASL) MRI has been successfully applied to measure gray matter (GM) perfusion in vivo, accurate detection of white matter (WM) perfusion has proven difficult. Reported literature values are not consistent with each other or with perfusion measured with other modalities. In this work, the cause of these inconsistencies is investigated. The results suggest that WM perfusion values are substantially affected by the limited image resolution and by signal losses caused by the long transit times in WM, which significantly affect the label. From gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA) bolus-tracking experiments (N=6), it is estimated that the transit time can be several seconds long in deep WM. Furthermore, simulations show that even at a spatial resolution of 7 microl voxel size, contamination by the GM signals can exceed 40% of the actual WM signal. From 10-min long flow-sensitive alternating inversion recovery ASL (FAIR-ASL) measurements at 3T in normal subjects (N=7), using highly sensitive detectors, it is shown that single-voxel (7 mul) deep WM perfusion values have an signal-to-noise ratio (SNR) less than 1. The poor sensitivity and heterogeneous transit time limit the applicability of ASL for measurement of perfusion in WM. PMID- 18383291 TI - Reducing correlated noise in fMRI data. AB - The sensitivity of functional MRI (fMRI) in detecting neuronal activation is dependent on the relative levels of signal and noise in the time-series data. The temporal noise level within a single voxel is generally substantially higher than the intrinsic NMR (thermal) noise, and the noise is often correlated between voxels. This work introduces and evaluates a method that allows fMRI sensitivity improvement by reduction of these correlated noise sources. The method allows model-free estimation of the correlated noise from brain regions not activated by the functional paradigm using a short (1-2 min) reference scan. A single regressor representing this noise-source estimate is added to the design matrix used in the data analysis. Results obtained from five volunteers show an average t-score improvement of 11.3% and a 24.2% increase in the size of the activated area. PMID- 18383292 TI - Multinuclear NMR investigation of probe construction materials at 9.4T. AB - This work investigates probe construction materials for their signal contribution to ultrashort echo time spectroscopy and imaging. (1)H, (13)C, and (31)P spectra were obtained at a field strength of 9.4 T for 16 materials considered for use in probe and holder design and construction. Four of the materials were found to be suited for the construction of NMR probes, housing of RF coils, and holders for in vivo experiments. PMID- 18383293 TI - Observation of microscopic diffusion anisotropy in the spinal cord using double pulsed gradient spin echo MRI. AB - A double-pulsed gradient spin echo (d-PGSE) filtered MRI sequence is proposed to detect microscopic diffusion anisotropy in heterogeneous specimen. The technique was developed, in particular, to characterize local microscopic anisotropy in specimens that are macroscopically isotropic, such as gray matter. In such samples, diffusion tensor MRI (DTI) produces an isotropic or nearly isotropic diffusion tensor despite the fact that the medium may be anisotropic at a microscopic length scale. Using d-PGSE filtered MRI, microscopic anisotropy was observed in a "gray matter" phantom consisting of randomly oriented tubes filled with water, as well as in fixed pig spinal cord, within a range of b-values that can be readily achieved on clinical and small animal MR scanners. These findings suggest a potential use for this new contrast mechanism in clinical studies and biological research applications. PMID- 18383294 TI - Improved image reconstruction for partial Fourier gradient-echo echo-planar imaging (EPI). AB - The partial Fourier gradient-echo echo planar imaging (EPI) technique makes it possible to acquire high-resolution functional MRI (fMRI) data at an optimal echo time. This technique is especially important for fMRI studies at high magnetic fields, where the optimal echo time is short and may not be achieved with a full Fourier acquisition scheme. In addition, it has been shown that partial Fourier EPI provides better anatomic resolvability than full Fourier EPI. However, the partial Fourier gradient-echo EPI may be degraded by artifacts that are not usually seen in other types of imaging. Those unique artifacts in partial Fourier gradient-echo EPI, to our knowledge, have not yet been systematically evaluated. Here we use the k-space energy spectrum analysis method to understand and characterize two types of partial Fourier EPI artifacts. Our studies show that Type 1 artifact, originating from k-space energy loss, cannot be corrected with pure postprocessing, and Type 2 artifact can be eliminated with an improved reconstruction method. We propose a novel algorithm, that combines images obtained from two or more reconstruction schemes guided by k-space energy spectrum analysis, to generate partial Fourier EPI with greatly reduced Type 2 artifact. Quality control procedures for avoiding Type 1 artifact in partial Fourier EPI are also discussed. PMID- 18383295 TI - Cross-validation-based kernel support selection for improved GRAPPA reconstruction. AB - The extended version of the generalized autocalibrating partially parallel acquisition (GRAPPA) technique incorporates multiple lines and multiple columns of measured k-space data to estimate missing data. For a given accelerated dataset, the selection of the measured data points for fitting a missing datum (i.e., the kernel support) that provides optimal reconstruction depends on coil array configuration, noise level in the acquired data, imaging configuration, and number and position of autocalibrating signal lines. In this work, cross validation is used to select the kernel support that best balances the conflicting demands of fit accuracy and stability in GRAPPA reconstruction. The result is an optimized tradeoff between artifacts and noise. As demonstrated with experimental data, the method improves image reconstruction with GRAPPA. Because the method is simple and applied in postprocessing, it can be used with GRAPPA routinely. PMID- 18383296 TI - Self-calibrating GRAPPA operator gridding for radial and spiral trajectories. AB - Self-calibrating GRAPPA operator gridding (GROG) is a method by which non Cartesian MRI data can be gridded using spatial information from a multichannel coil array without the need for an additional calibration dataset. Using self calibrating GROG, the non-Cartesian datapoints are shifted to nearby k-space locations using parallel imaging weight sets determined from the datapoints themselves. GROG employs the GRAPPA Operator, a special formulation of the general reconstruction method GRAPPA, to perform these shifts. Although GROG can be used to grid undersampled datasets, it is important to note that this method uses parallel imaging only for gridding, and not to reconstruct artifact-free images from undersampled data. The innovation introduced here, namely, self calibrating GROG, allows the shift operators to be calculated directly out of the non-Cartesian data themselves. This eliminates the need for an additional calibration dataset, which reduces the imaging time and also makes the GROG reconstruction more robust by removing possible inconsistencies between the calibration and non-Cartesian datasets. Simulated and in vivo examples of radial and spiral datasets gridded using self-calibrating GROG are compared to images gridded using the standard method of convolution gridding. PMID- 18383297 TI - Anatomically constrained reconstruction from noisy data. AB - Noise is a major concern in many important imaging applications. To improve data signal-to-noise ratio (SNR), experiments often focus on collecting low-frequency k-space data. This article proposes a new scheme to enable extended k-space sampling in these contexts. It is shown that the degradation in SNR associated with extended sampling can be effectively mitigated by using statistical modeling in concert with anatomical prior information. The method represents a significant departure from most existing anatomically constrained imaging methods, which rely on anatomical information to achieve super-resolution. The method has the advantage that less accurate anatomical information is required relative to super resolution approaches. Theoretical and experimental results are provided to characterize the performance of the proposed scheme. PMID- 18383298 TI - Fast parallel spiral chemical shift imaging at 3T using iterative SENSE reconstruction. AB - Spiral chemical shift imaging (CSI) is a fast CSI technique that simultaneously encodes 1D spectral and 2D spatial information. Therefore, it potentially allows one to perform a 2D-CSI experiment in a single shot. However, for most applications, limitations on maximum gradient strength and slew rate make multiple excitations necessary in order to achieve a desired spectral bandwidth. In this work we reduce the number of spatial interleaves and, hence, the minimum total measurement time of spiral CSI by using an iterative sensitivity encoding reconstruction algorithm which utilizes complementary spatial encoding afforded by the spatially inhomogeneous sensitivity profiles of individual receiver coils. The performance of the new method was evaluated in phantom and in vivo experiments. Parallel spiral CSI produced maps of brain metabolites similar to those obtained using conventional gridding reconstruction of the fully sampled data with only a small decrease in time-normalized signal-to-noise ratio and a small increase in noise for higher acceleration factors. PMID- 18383299 TI - Magnetic resonance imaging of neuronal and glial swelling as an indicator of function in cerebral tissue slices. AB - Here we demonstrate a new basis of signal change in magnetic resonance imaging (MRI) related to neuronal function, independent of blood oxygenation or flow. Time series MRI data acquired from living, superfused brain slices of adult rats revealed that the signal intensity reversibly increased with depolarization evoked by briefly elevating extracellular K(+). This was presumably a consequence of increased tissue water in the intracellular compartment. Reversible increases in light transmittance (LT) demonstrating a similar time course in response to K(+) elevation supported cellular swelling as generating the MRI signal intensity changes. This was confirmed by reversibly swelling cells in the slice under hypoosmotic challenge, which increased both MRI and LT signals with an identical time course. Conversely, shrinking cells under hyperosmotic challenge reversibly decreased the MRI and LT signals. We propose that specific MRI of neuronal function (fMRI) signals detected under identical parameters during predominantly proton-density-weighted fMRI of the spinal cord can now be explained by neuronal and glial swelling in activated central nervous system (CNS) regions. These observations demonstrate the biophysical basis of the fMRI contrast mechanism that has been termed "signal enhancement by extravascular water protons," or SEEP. PMID- 18383300 TI - Radial single-shot STEAM MRI. AB - Rapid MR imaging using the stimulated echo acquisition mode (STEAM) technique yields single-shot images without any sensitivity to resonance offset effects. However, the absence of susceptibility-induced signal voids or geometric distortions is at the expense of a somewhat lower signal-to-noise ratio than EPI. As a consequence, the achievable spatial resolution is limited when using conventional Fourier encoding. To overcome the problem, this study combined single-shot STEAM MRI with radial encoding. This approach exploits the efficient undersampling properties of radial trajectories with use of a previously developed iterative image reconstruction method that compensates for the incomplete data by incorporating a priori knowledge. Experimental results for a phantom and human brain in vivo demonstrate that radial single-shot STEAM MRI may exceed the resolution obtainable by a comparable Cartesian acquisition by a factor of four. PMID- 18383301 TI - Water chemical shift in 1H NMR of red cells: effects of pH when transmembrane magnetic susceptibility differences are low. AB - The (1)H magic angle spinning (MAS) NMR spectrum of water in erythrocyte suspensions shows peaks from each of the intracellular and extracellular water pools. The splitting is a true chemical shift and is brought about by the elimination of water exchange under MAS conditions due to physical separation of the two water populations. The size of the chemical shift difference is determined by the concentration of intracellular protein affecting the average extent of hydrogen bonding of water. We present here a model of the chemical shift behavior for water in erythrocytes under normal high-resolution NMR conditions based on results from MAS experiments on these cells exposed to different pH and osmotic conditions. The model accurately predicts the chemical shift of water for a static sample, and the results demonstrate that in high resolution NMR experiments the chemical shift of water will appear to be invariant if differences in magnetic susceptibility across the cell membrane are minimal (<10% of the magnetic susceptibility of water). Thus, changes in the shape and chemical shift of the water resonance are not due to pH changes in the physiological range. The findings are fundamental to an interpretation of the mechanism of chemical shift effects on the water resonance that may occur in functional MRI. PMID- 18383303 TI - Do hydrophobic interactions exist in the gas phase? PMID- 18383302 TI - In vivo quantification of T1, T2, and apparent diffusion coefficient in the mouse retina at 11.74T. AB - MRI has recently been used for noninvasive examination of retinal structure and function in rats and cats. However, the advantages of quantitative high resolution MRI of retina from mice have not yet been explored. In the present study, T(1) and T(2) relaxation time constants and the directional apparent diffusion coefficient (ADC) in the retina of C57/BL6 mice were measured. Three MR detected retina layers and a MR-detected choroid layer were observed on both T(1) and T(2)-weighted images at an image resolution of 47 x 47 x 400 microm(3). The significantly higher ADC parallel to than that perpendicular to the optic nerve in the MR-detected outer retina layer at the central retina reflects the known cellular organization of the photoreceptor cells. This study establishes, for the first time, normative metrics of T(1), T(2), and ADC of the mouse retina. These MR parameters are expected to be useful in future evaluation of developmental and pathological alterations of retinal cell layers in mice. PMID- 18383304 TI - Fractionated Feridex and positive contrast: in vivo MR imaging of atherosclerosis. AB - Macrophages have been identified as a critical factor in the pathogenesis of atherosclerosis. Ultrasmall iron oxide particles (USPIOs) have been used to passively target intraplaque macrophages. For dextran-based USPIOs, uptake into macrophages may be modulated by particle size. The aim of the current study was to test the efficacy of fractionated Feridex with respect to macrophage uptake in atherosclerotic rabbits. Fractionation of Feridex resulted in a 15-nm USPIO that exhibited a blood half-life of 15.9 h and liver retention of 6.4%. Blood clearance and liver retention of Feridex was 0.46 h and 60%, following administration of 4.8 mg Fe/kg Feridex. Atherosclerotic rabbits were administered 0.5 or 4.8 mg Fe/kg dosages of either fractionated Feridex or Feridex. MRI was performed at 1.5T over a 24-h time period postinjection. Perls and RAM-11 staining was performed to identify iron deposition. MRI showed a dose-dependent signal loss using conventional gradient echo (GRE) sequences following administration of fractionated Feridex. Even at low dose, significant signal loss was observed that correlated with histology. No signal attenuation or iron deposition was observed in the vessel wall of rabbits administered Feridex. Results of this study suggest that it may be possible to optimize USPIOs for intraplaque macrophage detection. PMID- 18383305 TI - Visualization of the coherence of the principal diffusion orientation: an eigenvector-based approach. AB - A novel method for spatially mapping anisotropy/orientation coherence of the eigenvector is presented. By using an eigenvector-based approach, an intervoxel diffusion coherence (IVDC) index was used to quantify the coherence of the principal diffusion directions within a voxel neighborhood. This method may allow reconstruction of a whole brain map to be used for diagnostic purposes. The IVDC index is calculated by a scatter matrix-based method in a voxel-wise manner. A simulation was performed using two fiber populations crossing at various separation angles. We demonstrate that the IVDC index was more sensitive than fractional anisotropy (FA) to changes in separation between the fibers under a noise-free condition. Diffusion-tensor images of six healthy volunteers were acquired on a 3.0T MR imager. The FA, coherence index, and IVDC were then calculated. The results showed that IVDC improved the contrast in several brain areas including thalamus, middle cerebral peduncle, and pons. We therefore conclude that the IVDC index provides reliable and complementary information on water diffusion in the brain. It may be useful in white matter tractography, especially to determine the termination point of a trajectory. PMID- 18383306 TI - Quantitative mapping of ventilation-perfusion ratios in lungs by 19F MR imaging of T1 of inert fluorinated gases. AB - A new method is presented for quantitative mapping of ventilation-to-perfusion ratios (V(A)/Q) in the lung: MRI of the (19)F longitudinal relaxation time (T(1)) of an inert fluorinated gas at thermal polarization. The method takes advantage of the dependence of the (19)F T(1) on the local SF(6) partial pressure, which depends on the local value of V(A)/Q. In contrast to hyperpolarized noble gases, with very long T(1)s, the T(1) of SF(6) in mammal lungs is 0.8-1.3 ms. Thus, rapid signal averaging overcomes the low thermal equilibrium polarization. T(1) imaging of a phantom consisting of four different SF(6)/air mixtures with known T(1) values validates the modified Look-Locker T(1) imaging sequence. To demonstrate the method in vivo, partial obstruction of the left bronchus was attempted in three rats; 3D free induction decay (FID)-projection T(1) images (2 mm isotropic resolution) revealed obstructed ventilation in two of the animals. In those images, approximately 1700 lung voxels contained sufficient SF(6) for analysis and T(1) was determined in each voxel with a standard error of 8-10%. For comparison, independent V(A)/Q images of the same animals were obtained using a previously described SF(6) MRI technique, and good agreement between the two techniques was obtained. Relative to the previous technique the resolution achieved using the T(1) method is lower (for similar V(A)/Q precision and imaging time); however, the T(1) method offers the potential advantages of eliminating the need for image coregistration and allowing patients with impaired lung function to breathe a 70% O(2) gas mixture during the entire imaging procedure. PMID- 18383307 TI - Accelerated whole-heart 3D CSPAMM for myocardial motion quantification. AB - Myocardial tissue tagging using complementary spatial modulation of magnetization (CSPAMM) allows detailed assessment of myocardial motion. To capture the complex 3D cardiac motion pattern, multiple 2D tagged slices are usually acquired in different orientations. These approaches are prone to slice misregistration and associated with long acquisition times. In this work, a fast method for acquiring 3D CSPAMM data is proposed that allows measuring deformation of the whole heart in three breath-holds of 18 heartbeats duration each. Three acquisitions are sequentially performed with line tag preparation in each orthogonal direction. Measurement acceleration is achieved by applying localized tagging preparation and a hybrid multishot, segmented echo-planar imaging sequence. Five healthy volunteers and five patients with myocardial infarction were measured. Midwall contours were tracked throughout the cardiac cycle with an enhanced variant of the harmonic phase (HARP) technique. Circumferential shortening at end-systole ranged from 14.1% (base) to 20.1% (apex) in healthy subjects. Hypokinetic regions in patients corresponded well with regions exhibiting hyperenhancement after contrast injection. Time to maximum circumferential shortening varied more significantly over the left ventricle in patients than in volunteers (P<0.01). The proposed measurement scheme was well tolerated by patients and holds considerable potential to investigate cardiac mechanics in various diseases. PMID- 18383308 TI - Mass spectrometric analysis of the marine lipophilic biotoxins pectenotoxin-2 and okadaic acid by four different types of mass spectrometers. AB - The performances of four different mass spectrometers [triple-quadrupole (TQ), time-of-flight (ToF), quadrupole ToF (Q-ToF) and ion trap (IT)] for the detection of the marine lipophilic toxins pectenotoxin-2 (PTX2) and okadaic acid (OA) were investigated. The spectral data obtained with the different mass spectrometric analyzers were used to propose fragmentation schemes for PTX2 in the positive electrospray mode and for OA in the negative electrospray mode. TQ data were used to obtain product ions, while ToF and Q-ToF-MS produced accurate mass data of the precursor ion and product ions, respectively. IT data provided a better understanding of the fragmentation pathways using MS(n) experiments. With respect to analytical performance, all four mass analyzers showed a good linearity (R(2) > 0.97) and repeatability (CV < 20%). Detection limits (LoDs) (S/N = 3) were the lowest on triple-quad MS: 12.2 and 2.9 pg on-column for PTX2 and OA, respectively. PMID- 18383310 TI - Cytometric fingerprinting: quantitative characterization of multivariate distributions. AB - Recent technological advances in flow cytometry instrumentation provide the basis for high-dimensionality and high-throughput biological experimentation in a heterogeneous cellular context. Concomitant advances in scalable computational algorithms are necessary to better utilize the information that is contained in these high-complexity experiments. The development of such tools has the potential to expand the utility of flow cytometric analysis from a predominantly hypothesis-driven mode to one of discovery, or hypothesis-generating research. A new method of analysis of flow cytometric data called Cytometric Fingerprinting (CF) has been developed. CF captures the set of multivariate probability distribution functions corresponding to list-mode data and then "flattens" them into a computationally efficient fingerprint representation that facilitates quantitative comparisons of samples. An experimental and synthetic data were generated to act as reference sets for evaluating CF. Without the introduction of prior knowledge, CF was able to "discover" the location and concentration of spiked cells in ungated analyses over a concentration range covering four orders of magnitude, to a lower limit on the order of 10 spiked events in a background of 100,000 events. We describe a new method for quantitative analysis of list mode cytometric data. CF includes a novel algorithm for space subdivision that improves estimation of the probability density function by dividing space into nonrectangular polytopes. Additionally it renders a multidimensional distribution in the form of a one-dimensional multiresolution hierarchical fingerprint that creates a computationally efficient representation of high dimensionality distribution functions. CF supports both the generation and testing of hypotheses, eliminates sources of operator bias, and provides an increased level of automation of data analysis. PMID- 18383309 TI - Nine-color flow cytometry for accurate measurement of T cell subsets and cytokine responses. Part II: Panel performance across different instrument platforms. AB - Cellular immune responses elicited by vaccination are complex and require polychromatic analysis to accurately characterize the phenotype and function of rare, responding cells. Technical challenges and a lack of instrument standardization between research sites have limited the application of polychromatic cytometry in multicenter clinical trials. Two previously developed six-color T cell subset immunophenotyping reagent panels deliberately designed to accommodate three additional low frequency functional measurements were compared for their reproducibility of staining across three different flow cytometers. We repeatedly measured similar T cell subset frequencies between the two reagent panels and across the three different cytometers. Spectral overlap reduced sensitivity in two of the three open measurement channels (PE [IL-2] and APC [IFN gamma]) for one reagent combination, particularly in subsets with low cytokine expression. There was no significant interassay variation for measurements across instrument platforms. Careful panel design will identify reagent combinations that minimize spectral spillover into channels reserved for cytokine measurement and comparable results can be achieved using different cytometers, however, it is important to establish standardized quality control procedures for each instrument to minimize variation between cytometers. PMID- 18383311 TI - Mixture modeling approach to flow cytometry data. AB - Flow Cytometry has become a mainstay technique for measuring fluorescent and physical attributes of single cells in a suspended mixture. These data are reduced during analysis using a manual or semiautomated process of gating. Despite the need to gate data for traditional analyses, it is well recognized that analyst-to-analyst variability can impact the dataset. Moreover, cells of interest can be inadvertently excluded from the gate, and relationships between collected variables may go unappreciated because they were not included in the original analysis plan. A multivariate non-gating technique was developed and implemented that accomplished the same goal as traditional gating while eliminating many weaknesses. The procedure was validated against traditional gating for analysis of circulating B cells in normal donors (n = 20) and persons with Systemic Lupus Erythematosus (n = 42). The method recapitulated relationships in the dataset while providing for an automated and objective assessment of the data. Flow cytometry analyses are amenable to automated analytical techniques that are not predicated on discrete operator-generated gates. Such alternative approaches can remove subjectivity in data analysis, improve efficiency and may ultimately enable construction of large bioinformatics data systems for more sophisticated approaches to hypothesis testing. PMID- 18383312 TI - Accurate classification of MLH1/MSH2 missense variants with multivariate analysis of protein polymorphisms-mismatch repair (MAPP-MMR). AB - Lynch syndrome, also known as hereditary nonpolyposis colon cancer (HNPCC), is the most common known genetic syndrome for colorectal cancer (CRC). MLH1/MSH2 mutations underlie approximately 90% of Lynch syndrome families. A total of 24% of these mutations are missense. Interpreting missense variation is extremely challenging. We have therefore developed multivariate analysis of protein polymorphisms-mismatch repair (MAPP-MMR), a bioinformatic algorithm that effectively classifies MLH1/MSH2 deleterious and neutral missense variants. We compiled a large database (n>300) of MLH1/MSH2 missense variants with associated clinical and molecular characteristics. We divided this database into nonoverlapping training and validation sets and tested MAPP-MMR. MAPP-MMR significantly outperformed other missense variant classification algorithms (sensitivity, 94%; specificity, 96%; positive predictive value [PPV] 98%; negative predictive value [NPV], 89%), such as SIFT and PolyPhen. MAPP-MMR is an effective bioinformatic tool for missense variant interpretation that accurately distinguishes MLH1/MSH2 deleterious variants from neutral variants. PMID- 18383313 TI - Translational research: Whither the ORS? AB - There is a hue and cry from the public, policy makers and funding agencies about the primacy of research translation. The federal government has responded with the NIH Roadmap for Medical Research initiative. It is proposed that the Orthopaedic Research Society (ORS) initiate its own translational roadmap along themes that parallel the NIH Roadmap: the formation of interdisciplinary teams; the focus on specific musculoskeletal diseases; cross disciplinary education in essential translational research skills; fostering translational career pathways; and providing critical fora for translational research interactions--a "translational interface." By its very structure, the ORS is an organization that is uniquely poised to help its members respond to the demands for research translation. The roadmap outlined around these five themes is intended to make the ORS even more relevant in our professional lives and to transform the ORS into an organization that is a true catalyst for improving the musculoskeletal health of the public. PMID- 18383314 TI - Casimer T. Grabowski: 1927-2007. PMID- 18383315 TI - Results of the negative control chemical allyl alcohol in the 15-day intact adult male rat screening assay for endocrine activity. AB - Development, standardization, and validation of methods to assess the potential of chemicals to disrupt hormonal homeostasis have been the focus of considerable research efforts over the past 10 years. As part of our validation effort, we evaluated the specificity of the 15-day intact adult male rat assay, using a negative control chemical, allyl alcohol, a known hepatotoxicant that was not expected to induce endocrine effects. Male rats were dosed for 15 days via oral gavage with 0, 10, 30, 40, or 50 mg/kg/day allyl alcohol. The endpoints evaluated included final body and organ weights, serum hormone concentrations, and a limited histopathology assessment. No mortality or adverse clinical signs were observed. Mean final body weight for rats in the 50-mg/kg/day dose group was decreased to 90% of control. Mean relative liver weights were increased at 40 and 50 mg/kg/day (115% and 117% of control, respectively). Serum testosterone and DHT concentrations were statistically significantly decreased at 50 mg/kg/day (72% of control). Serum prolactin concentrations were statistically significantly decreased at 40 mg/kg/day (58% of control), but not at 50 mg/kg/day. There were no effects on the other endpoints evaluated. Consistent with previous guidance for interpreting the 15-day intact adult male rat assay, histological and weight changes of target organs were given a higher weight-of-evidence than changes in serum hormone concentrations alone. Therefore, with only minimal changes in serum hormone concentrations and no effects on organ weights or microscopic alterations, the results of allyl alcohol in the 15-day intact adult male rat assay were considered negative and consistent with the predicted results. PMID- 18383316 TI - Nine-color flow cytometry for accurate measurement of T cell subsets and cytokine responses. Part I: Panel design by an empiric approach. AB - Polychromatic flow cytometry offers the unprecedented ability to investigate multiple antigens per cell. Unfortunately, unwanted spectral overlaps and compensation problems increase when more than four colors are used, but these problems can be minimized if staining combinations are chosen carefully. We used an empiric approach to design, test and identify six-color T cell immunophenotyping reagent panels that can be expanded to include three or more functional or other markers in the FITC, PE, and APC channels without significant spectral limitations. Thirty different six-color T cell surface antigen reagent panels were constructed to identify major T cell subsets and maturational subtypes as defined by CCR7 and CD45RA expression, while excluding monocytes, B and non-viable cells. Staining performance of each panel was compared on cryopreserved cells from a single healthy donor recorded on a multiparameter cell sorter. Ten of the thirty reagent panels offered reliable resolution of T cell major and maturational surface markers. Of these, two panels were selected that showed the least spectral overlap and resulting background increase in the FITC, PE, and APC channels. These channels were left unoccupied for inclusion of additional phenotypic or functional markers, such as cytokines. Careful reagent titration and testing of multiple candidate panels are necessary to ensure quality results in multiparametric measurements. PMID- 18383317 TI - De novo maintenance therapy with denileukin diftitox (Ontak) in a patient with peripheral T-cell lymphoma is associated with prolonged remission. AB - Peripheral T-cell lymphoma (PTCL) is an aggressive form of non-Hodgkin's lymphoma (NHL), associated with poor prognosis and without standard approach to treatment. Denileukin diftitox (Ontak) is a synthetic fusion protein combining the receptor binding domain of interleukin-2 to the enzymatically active portion of diphtheria toxin. While approved for the treatment of cutaneous T-cell lymphoma, it has demonstrated activity in non-Hodgkin's lymphomas of both T-cell and B-cell origin. This report documents the first case of de novo maintenance therapy with denileukin diftitox sustaining an ongoing complete response at the molecular level for 2 years in a patient with PTCL. PMID- 18383318 TI - Plasma concentrations of asymmetric dimethylarginine, an endogenous nitric oxide synthase inhibitor, are elevated in sickle cell patients but do not increase further during painful crisis. AB - Plasma concentrations of asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, are elevated in the clinically asymptomatic state of sickle cell disease (SCD). However, the role of ADMA during vaso-occlusive complications has not been defined. ADMA concentrations were determined in HbSS (n = 43) and HbSC (n = 25) patients with healthy blood donors (HbAA) as controls. In the clinically asymptomatic state ADMA concentrations were elevated in sickle cell patients as compared to healthy controls (HbSS 0.70 micromol/L, HbSC 0.54 micromol/L, HbAA 0.39 micromol/L) (P < 0.001). Yet plasma ADMA concentrations did not increase further at presentation with a painful crisis implicating no role of primary importance during vaso-occlusive crises. PMID- 18383319 TI - Graft rejection after hematopoietic cell transplantation with nonmyeloablative conditioning. AB - Graft rejection after hematopoietic cell transplantation (HCT) with nonmyeloablative conditioning is a rare but serious clinical problem. Graft rejection and salvage therapy in eight patients in a retrospective analysis of 124 consecutive patients is reported. The patients were conditioned with low-dose fludarabine and total body irradiation (TBI). The association of pretransplantation risk factors with rejection and the effect of chimerism and graft-versus-host disease on rejection were analyzed. Overall survival (OS) and progression free survival (PFS) were compared between patients with and without rejection. Retransplantation was performed with increased TBI conditioning for all patients, and with increased mycophenolate mofetil doses for recipients with HLA-identical sibling donors. No known pretransplantation risk factors were confirmed in this study. Rejection episodes were unevenly distributed over time. The storage temperature of the apheresis products was identified as a risk factor for rejection. Storage of the apheresis products at 5 degrees C diminished the risk of rejection. Low donor T cell chimerism at Day +14 significantly increased the risk of rejection. Seven patients were retransplanted. All but one engrafted successfully, but with decreased OS and PFS. Two patients received pentostatin infusion prior to donor lymphocyte infusions in unsuccessful attempts at reversing rejection. Storage temperature and donor chimerism had a significant effect on rejection. Following rejection, patients are at greater risk of dying from infections and progression/relapse of their malignancy. Retransplantation is feasible and well tolerated after HCT with nonmyeloablative conditioning and should be performed without delay in patients with imminent and manifest graft rejection. PMID- 18383320 TI - Transfusion-related acute lung injury following intravenous anti-D administration in an adolescent. AB - Transfusion-related acute lung injury (TRALI) is associated with administration of all plasma containing blood products. We present a 14-year-old adolescent diagnosed with idiopathic thrombocytopenic purpura who developed acute respiratory insufficiency compatible with TRALI within 5 hr following intravenous anti-D. Full blown noncardiogenic pulmonary edema was noted after 9 hr. Mechanical ventilation was not required and the patient made a full recovery after 36 hr. Analysis of the anti-D preparation revealed reactivity against the neutrophil FcgammaRIIIb. A postinfusion serum sample contained antibodies against class I human HLA-A11 antigen. Clinicians should consider TRALI in patients developing unexplained dyspnea after receiving intravenous anti-D. PMID- 18383321 TI - Bone marrow glycophorin-positive erythroid cells of myelodysplastic patients responding to high-dose rHuEPO therapy have a different gene expression pattern from those of nonresponders. AB - The main clinical problems of low-risk patients with myelodysplastic syndromes (MDS), as defined by the International Prognostic Scoring System, are infections and the need for frequent transfusions due to ineffective myelopoiesis and peripheral blood cytopenia. Promising results in treating MDS-related anemia have been obtained using high-dose recombinant human erythropoietin (rhEPO). To evaluate the molecular basis of the response to rhEPO, we used commercially available macro-arrays to investigate gene expression profiles in the glycophorin expressing (Gly+) bone marrow (BM) erythroid cells of five responders (ERs) and five non-responders (ENRs) to rhEPO treatment. The cells were separated by means of positive selection using an immunomagnetic procedure, after which flow cytometry showed that their purity was more than 97% in all cases. The array data were validated by means of real time RT-PCR. The results showed that the genes responsible for proliferation/differentiation and DNA repair/stability were repressed in the BM Gly+ erythroid cells of the ENRs, but almost normally expressed in the ERs. Furthermore, the expression of genes involved in signal transduction suggested that the activity of the MAPK signaling pathway is inhibited in ERs. The different gene expression profiles of ERs and ENRs may provide a basis for early gene testing as a means of predicting the response to rhEPO of MDS patients with low endogenous EPO levels. PMID- 18383322 TI - Lipoprotein-associated phospholipase A2 and risk of venous thrombosis in older adults. AB - Lipoprotein-associated phospholipase A2 (Lp-PLA2) is an enzyme involved in inflammation and platelet function. Inherited deficiency and elevated levels are associated with atherosclerosis. Given potential common etiologies of atherosclerosis and venous thrombosis (VT), we hypothesized that low and high Lp PLA2 would be associated with VT risk. Lp-PLA(2) mass and activity were measured in baseline samples of Cardiovascular Health Study participants (5,888 men and women age > or =65), excluding 354 reporting pre-baseline VT. The study endpoint was VT unrelated to cancer after 11.6 years follow-up. Hazard ratios were estimated using Cox proportional hazard models, adjusting for age, race, sex, and body-mass index. With 129 cases of VT, there was no association of Lp-PLA2 activity with risk. Adjusted hazard ratios were 1.19 (CI 0.62, 2.29) and 0.87 (CI 0.43, 1.76) for the lowest and highest decile, respectively, compared to the 10 25th percentile. Corresponding hazard ratios for Lp-PLA2 mass were 1.63 (CI 0.79, 3.34) and 1.33 (CI 0.61, 2.87). Results were robust to several definitions of low or high Lp-PLA2. While the association of Lp-PLA(2) levels with arterial disease events implies a role for this enzyme in atherogenesis, our findings suggest that it is not prothrombotic. PMID- 18383323 TI - Bone marrow fibrosis and gelatinous atrophy associated with acute Epstein-Barr virus infection. PMID- 18383324 TI - Polymorphisms of the human platelet alloantigens HPA-1, HPA-2, HPA-3, and HPA-4 in ischemic stroke. AB - Polymorphism in human platelet antigen (HPA)-1 and HPA-3 (GPIIb/IIIa), HPA-2 (GPIb/IX), HPA-4 (GPIIIa), and HPA-5 (GPIa/IIa) was investigated in 329 stroke patients and 444 matched control subjects. HPA genotyping was done by PCR-SSP method. Lower HPA-1a (P < 0.001) and higher HPA-1b (P < 0.001) allele frequencies were seen in patients than control subjects, and homozygosity for HPA-1b (P < 0.001) alleles was more prevalent in stroke cases than in controls. The allele and genotype distributions of the other HPA polymorphic variants were similar between cases and controls. Select HPA combined genotypes comprising the 2121 (Pc = 0.008) and 2221 (Pc = 0.018) genotypes, which were positively associated, and the 1111 (Pc < 0.001), which was negatively associated with stroke, thereby conferred a disease susceptibility and protective nature to these genotype combinations. Multivariate analysis confirmed the negative association of the 1111 (P < 0.001) and the positive association of the 2121 (P = 0.017) combined genotypes with stroke, after adjustment for a number of covariates. This is the first evidence demonstrating differential association of the common 4 HPA gene variants and specific HPA genotype combinations with stroke. PMID- 18383325 TI - Longitudinal analysis of pulmonary function in adults with sickle cell disease. AB - Among adults with sickle cell disease (SCD), pulmonary complications are a leading cause of death. Yet, the natural history of lung function in adults with SCD is not well established. We conducted a retrospective cohort study of adults with SCD who had repeated pulmonary function tests performed over 20 years of age. Ninety-two adults were included in this cohort. Rate of decline in FEV(1) for men and women with SCD was 49 cc/year (compared with 20-26 cc/year in the general population). Further studies are needed to identify factors which impact the rate of lung function decline in adults with SCD. PMID- 18383326 TI - Neutrophilic dermatosis of the hands: an underrecognized hematological condition that may result in unnecessary surgery. PMID- 18383327 TI - The clinical and laboratory approach to the CD5/CD10 negative B-cell chronic lymphoproliferative leukemias. PMID- 18383328 TI - Long-term results of survival in patients with thalassemia major treated with bone marrow transplantation. AB - Allogeneic bone marrow transplantation (BMT) is the only available curative approach for thalassemia major, although long-term morbidity and mortality are not established. The aim of this study was to assess the long-term clinical and hematological results in children and adults with thalassemia major treated with BMT. We analyzed the outcome of 115 patients (median age 9 years, range 11 months to 28 years) with thalassemia major undergoing BMT from a related donor between 1983 and 2006. All patients received the same protocol, consisting of busulfan and cyclophoshamide as conditioning therapy and cyclosporin (CSA) alone or CSA and methotrexate for graft-versus-host disease (GvHD) prophylaxis. The cumulative probability of graft rejection was 6.7%. The transplant-related mortality at 1 year was 8.7%. The 20-year Kaplan-Meier estimate of overall survival and disease free survival was 89.2% and 85.7%, respectively. Ninety-nine patients out of 103 survivors were in excellent clinical and hematological conditions at last visit following a median follow-up of 15 years (range, 1-24 years) with the exception of two patients who had invalidating chronic GvHD. This study conducted with a large cohort of patients and covering a long period of observation time, showed BMT to be curative for the majority of patients with thalassemia major. The impact of long-term transplant-related sequelae was very limited. PMID- 18383329 TI - Pulmonary arterial hypertension and left-sided heart disease in sickle cell disease: clinical characteristics and association with soluble adhesion molecule expression. AB - Pulmonary hypertension (PH), a risk factor for mortality in sickle cell disease (SCD), has pathologic features of both pulmonary arterial hypertension (PAH) and PH related to left-sided heart disease (LHD) suggesting a link between these two entities. We hypothesized that both are characterized by endothelial dysfunction and increased adhesion molecule expression. SCD patients and normal volunteers underwent a screening questionnaire, echocardiogram, and blood donation for preparation of platelet-poor plasma. PAH was defined as a tricuspid regurgitant jet (TRJ) velocity > or =2.5 m/sec and/or the presence of isolated right ventricular hypertrophy or decreased systolic function. LHD was defined as either left-sided systolic/diastolic dysfunction or significant valvular disease. Plasma vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), P- and E-selectin, nitric oxide (NO(x)), erythropoietin, and vascular endothelial growth factor (VEGF) levels were assayed by enzyme-linked immunoassay. Forty-three percent of sickle cell anemia (HbSS) and 28% of hemoglobin SC disease (HbSC) disease patients had PAH. Additionally, 10-15% of SCD patients had LHD. VCAM-1 levels were significantly increased in HbSS patients compared with HbSC patients and normal volunteers. VCAM-1 and P-selectin levels correlated positively with TRJ velocity in HbSS patients (r = 0.45, P = 0.03, r = 0.2, P = 0.05, respectively). ICAM-1, E-selectin, NO(x), erythropoietin, and VEGF levels were similar across subject groups. PH is common in SCD and, at times, due to LHD. Increased VCAM-1 and P-selectin expression was associated with TRJ elevation regardless of etiology suggesting a similar effect on endothelial gene expression and possibly providing a pathologic link between PAH and PH related to LHD in SCD. PMID- 18383330 TI - Complications of beta-thalassemia intermedia: a 12-year Lebanese experience. PMID- 18383331 TI - Procoagulant activity in children with community acquired pneumonia, pleural effusion and empyema. AB - In patients with community-acquired pneumonia (CAP), bacterial-cell-wall-derived fragments may induce the coagulation cascade. To contribute to the knowledge of underlying mechanisms, we have studied the fibrinolytic activity in children with CAP and parapneumonic effusions. PATIENTS AND METHODS: Twenty previously healthy children admitted to our Department with CAP were studied; with (n = 11) or without (n = 9) pleural effusion (PPE). We also investigated 10 children with empyema. In all children we analyzed coagulation and fibrinolytic parameters and compared the results to nine controls. RESULTS: Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were not significantly modified in the three groups as compared to controls (P = 0.975, P = 0.535, respectively). The fibrinogen levels were significantly increased in respect to the control group (P < 0.0001). The median values of D-dimer showed an increasing trend that was statistically significant: children with pneumonia 244 microg/L, with pneumonia and PPE 751 microg/L and with empyema 2003 microg/L, in respect to values (48 microg/L) of our control group (P < 0.0001). CONCLUSION: The results suggest that plasma level of D-dimer can give an additional contribution for the evaluation of the severity of CAP and its complications in children. PMID- 18383332 TI - Early lung disease in young children with primary ciliary dyskinesia. AB - Primary ciliary dyskinesia (PCD) is an autosomal recessive disease in which ciliary dysfunction leads to chronic lung, sinus, and middle ear disease. PCD is often not diagnosed until late childhood due to its presumed rarity and the technical expertise necessary for diagnosis; as such, little is known about lung disease in young children with PCD. We report on 3 young children with PCD who had evidence of lung disease on infant pulmonary function testing, bronchoscopy, and/or computed tomography (CT) of the chest before 3 years of age. PMID- 18383333 TI - Influence of beta2-adrenoceptor polymorphisms on the response to chronic use of albuterol in asthmatic children. AB - BACKGROUND: Several studies have suggested that after regular use of short acting beta2-agonists the bronchodilator effect of the drug may decline and this condition would be related to polymorphisms of the beta2-adrenergic receptor (beta2-AR). OBJECTIVE: To assess the frequency of beta2-AR polymorphisms in asthmatic children from Argentina, and to evaluate their influence on bronchodilator desensitization to albuterol over a 4-week treatment. METHODS: beta2-AR genotypes were determined in 117 children with asthma and 101 of them were under 4 weeks treatment with albuterol. Spirometric changes in FEV(1) were recorded at the beginning (day 1) and at the end of the study (day 30) and compared to genotypes at position 16 and 27 of the receptor. The frequency of the polymorphisms was calculated in all population. RESULTS: The presence of glutamine at position 27 (Gln27) was significantly more frequent in this Argentinean study population than in other Caucasian populations. The homozygosity for Gln27 polymorphism was associated to a desensitization of the receptor with a decline in the bronchodilator response to albuterol after chronic use. CONCLUSION: Gln27 polymorphism might be a marker for adverse clinical outcomes with chronic beta2-agonist exposure in children with asthma from Argentina. PMID- 18383334 TI - Diagnosing asthma in children: what is the role for methacholine bronchoprovocation testing? AB - OBJECTIVE: To determine whether measurement of airways responsiveness to methacholine can help physicians diagnose asthma in children. METHODS: Children from the 1995 Manitoba birth cohort were assessed by asthma specialists, had skin testing and measurement of airways responsiveness to methacholine (PC20). We selected children with doctor-diagnosed asthma and healthy children as controls (no asthma, no allergic rhinitis, negative skin tests). Sensitivities and specificities for asthma were calculated. Receiver operating curves were calculated to determine the best fit of the methacholine challenge as a diagnostic test. RESULTS: 640 children were assessed. Two hundred fifteen children with doctor diagnosed asthma and 197 healthy controls successfully completed a methacholine challenge. Airways hyperresponsiveness was a moderately sensitive and specific measure for the diagnosis of asthma in girls, whether atopic (sensitivity of 71% and specificity of 69% at PC20 < or = 4.0 mg/ml) or not (sensitivity of 77% and specificity of 53% at PC20 < or =8.0 mg/ml). Airways hyperresponsiveness was also helpful for the diagnosis of asthma in atopic boys (sensitivity of 67% and specificity of 75% at PC20 < or =2.0 mg/ml), but of absolutely no help in the diagnosis of asthma in nonatopic boys. CONCLUSION: Measurement of airways hyperresponsiveness to methacholine can be useful in children who are atopic and of some value in nonatopic girls. The presence or absence of airways hyperresponsiveness to methacholine is of no help for the diagnosis of asthma in nonatopic boys. Laboratory tests must be placed in context of the clinical assessment of children for asthma. PMID- 18383335 TI - Formulation and in vivo evaluation of chlorpropham (CIPC) oral formulations. AB - The objective of these studies was to examine the in vivo performance of oral formulations of chlorpropham (CIPC). In order to develop a new oral formulation several different solubilization techniques were evaluated, namely: cosolvents, surfactants, and complexing agents. The solubilization data indicated that a conventional solution formulation was not plausible. Two self-emulsifying drug delivery systems (SEDDS) were developed and evaluated for stability. Both SEDDS formulations were found to be chemically stable. In vivo analysis of a SEDDS formulation, a suspension formulation and an intravenous bolus dose was conducted in F344 rats. Pharmacokinetic analysis of the formulation data indicated that the SEDDS formulation provided only marginally better oral bioavailability compared to a suspension formulation. While SEDDS formulations often result in greater bioavailability this was not observed for CIPC. In vivo analysis indicate that CIPC results in a situation where the dissolution rate of CIPC from the suspension is not rate limiting, rather the absorption rate in the GI tract is rate-limiting. This paradigm is the result of CIPCs low melting point and the relatively small particle size of the suspension which facilitate the dissolution in the GI tract. PMID- 18383336 TI - Allosteric kinetics of human carboxylesterase 1: species differences and interindividual variability. AB - Esterified drugs such as imidapril, derapril, and oxybutynin hydrolyzed by carboxylesterase 1 (CES1) are extensively used in clinical practice. The kinetics using the CES1 substrates have not fully clarified, especially concerning species and tissue differences. In the present study, we performed the kinetic analyses in humans and rats in order to clarify these differences. The imidaprilat formation from imidapril exhibited sigmoidal kinetics in human liver microsomes (HLM) and cytosol (HLC) but Michaelis-Menten kinetics in rat liver microsomes and cytosol. The 2-cyclohexyl-2-phenylglycolic acid (CPGA) formation from oxybutynin were not detected in enzyme sources from rats, although HLM showed high activity. The kinetics were clarified to be different among species, tissues, and preparations. In individual HLM and HLC, there was large interindividual variability in imidaprilat (31- and 24-fold) and CPGA formations (15- and 9 fold). Imidaprilat formations exhibited Michaelis-Menten kinetics in HLM and HLC with high activity but sigmoidal kinetics in those with low activity. CPGA formations showed sigmoidal kinetics in high activity HLM but Michaelis-Menten kinetics in HLM with low activity. We revealed that the kinetics were different between individuals. These results could be useful for understanding interindividual variability and for the development of oral prodrugs. PMID- 18383337 TI - Assessment of presystemic and systemic intestinal availability of orally administered drugs using in vitro and in vivo data in humans: intestinal sulfation metabolism impacts presystemic availability much more than systemic availability of salbutamol, SULT1A3 substrate. AB - Although hepatic availability has been extensively studied to assess the oral bioavailability of drugs, intestinal availability has not, especially that related to conjugative metabolism (phase II metabolism). The present study assessed intestinal presystemic availability by integrating the reported metabolism data in vitro and in vivo of salbutamol, SULT1A3 substrate, in humans. Intrinsic clearance from each organ was calculated with the reported kinetic parameters for salbutamol sulfation metabolism in vitro. Then, presystemic (except intestine) and systemic organ availability and organ clearance were estimated by scaling up the in vitro data using biochemical and physiological data. Scaling factors used were one third, one and three times. Intestinal presystemic availability was calculated without assumptions of a well-stirred model. Presystemic availability of the intestine is much lower than systemic availability of the intestine. In addition, presystemic availability of the intestine is lower than presystemic availability of other organs, the liver or lung. SULT1A3 is expressed primarily in the intestine; therefore, it should be noted that intestinal metabolism affects the oral bioavailability of drugs, which are metabolized by SULT1A3. PMID- 18383338 TI - Pharmacokinetics of DS-96, an alkylpolyamine lipopolysaccharide sequestrant, in rodents. AB - The pharmacokinetics of DS-96, an N-alkylhomospermine analog designed to sequester bacterial lipopolysaccharides, has been determined in rodent species. The elimination half-life in mice and rats are about 400 and 500 min, respectively, with other PK parameters being quite similar in the two rodent species. Interestingly, the mouse intravenous plasma concentration time curves exhibit an apparent absorption phase. While the rat intravenous data did not exhibit a pronounced apparent absorption phase immediately following injection, plasma levels did increase between 10 and 30 min following an expected drop from time 0 to 5 min. The data are consistent with first-pass uptake, possibly by the lung, with back diffusion as a function of time. The observed C(max) values of 1.36 microg/mL in the mouse intraperitoneal model suggest that a plasma concentration of 0.5-1 microg/mL corresponds to complete protection for a 200 ng/animal dose of intraperitoneally administered LPS in the D-galactosamine primed model of endotoxin-induced lethality. PMID- 18383339 TI - Further evidence for a crucial role of resident endoneurial macrophages in peripheral nerve disorders: lessons from acrylamide-induced neuropathy. AB - Endoneurial macrophages are crucially involved in the pathogenesis of neuropathies. Historically, the macrophage response in neuropathies is believed to be of hematogenous origin. However, recent studies could demonstrate an intrinsic generation of the early macrophage response by resident endoneurial macrophages after traumatic nerve injury and in a model of hereditary neuropathy. We hypothesized that the local macrophage response might suffice to generate an appropriate macrophage response in mild neuropathies, supplemented by infiltrating macrophages only in severe nerve pathology. To clarify this assumption, we investigated the macrophage response in acrylamide-induced neuropathy as a model of a slowly progressive neuropathy with a defined onset. We induced the neuropathy in bone marrow chimeric mice carrying green fluorescent protein transgenic bone marrow, allowing the differentiation of resident (GFP(-)) and invading hematogenous endoneurial (GFP(+)) macrophages. Quantification of GFP(-) and GFP(+) endoneurial macrophages in the sciatic nerve revealed an increase only of resident macrophages in proximal parts, whereas in distal parts a minor additional influx of hematogenous macrophages was observed. The immunohistochemical profile of GFP(-) and GFP(+) macrophages was similar but distal GFP(-) macrophages were differentially activated than their GFP(+) counterparts. Characterization of CCR2-deficient mice revealed a function for this chemokine system in attracting hematogenous macrophages but not in generating the intrinsic macrophage response. In conclusion, we provide evidence for a role of resident macrophages in acrylamide-induced neuropathy. Resident endoneurial macrophages intrinsically generate the macrophage response in this slowly progressive neuropathy, which only becomes supplemented by hematogenous macrophages in distal areas of more pronounced damage. PMID- 18383340 TI - Increase of MCP-1 (CCL2) in myelin mutant Schwann cells is mediated by MEK-ERK signaling pathway. AB - Macrophages are critically involved in the pathogenesis of genetically caused demyelination, as it occurs in inherited demyelinating neuropathies. On the basis of the observation that upregulation of the Schwann cell-derived chemokine MCP-1 (CCL2) is a pathologically relevant mechanism for macrophage activation in mice heterozygously deficient for the myelin component P0 (P0+/-), we posed the question of the intracellular signaling cascade involved. By using western blot analysis of peripheral nerve lysates the MAP-kinases extracellular signal regulated kinase 1/2 (ERK1/2) and MAP kinase/ERK kinase 1/2 (MEK1/2) showed an early and constantly increasing activation in P0 mutants. Furthermore, in nerve fibers from the P0+/- mutants, Schwann cell nuclei were much more often positive for phosphorylated ERK1/2 than in nerve fibers from wild type mice. In vitro experiments using the MEK1/2-inhibitor CI-1040 decreased ERK1/2-phosphorylation and MCP-1 expression in a Schwann cell-derived cell line. Finally, systemic application of CI-1040 lead to a decreased ERK1/2-phosphorylation and substantially reduced MCP-1-production in peripheral nerves of P0+/- mutant mice. Our study identifies MEK1/2-ERK1/2 signaling as an important intracellular pathway that connects the Schwann cell mutation with the activation of pathogenetically relevant macrophages in the peripheral nerves. These findings may have important implications for the treatment of inherited peripheral neuropathies in humans. PMID- 18383341 TI - Regulation of prostaglandin E2 synthase expression in activated primary rat microglia: evidence for uncoupled regulation of mPGES-1 and COX-2. AB - Prostaglandin E2 (PGE2) is among the most important mediators involved in neuroinflammatory processes. The final step of its synthesis is regulated by enzymes termed prostaglandin E2 synthases (PGES). Three PGES are known, cytosolic (c)PGES, membrane-associated (m)PGES-1 and mPGES-2. The expression of mPGES-1 is induced by inflammatory stimuli such as lipopolysaccharide (LPS), interleukin (IL)-1beta, and tumor necrosis factor (TNF)-alpha. Although some roles of mPGES-1 have already been suggested, its function in the CNS and the signaling pathways involved in its upregulation are poorly understood. In this study, we examined the regulation of mPGES-1 in primary rat microglia and the signaling pathways involved in its expression. Whereas the expression of cPGES and mPGES-2 was not stimulated by LPS, low doses of LPS (0.1-1 ng/mL) sufficiently stimulated mPGES-1 mRNA expression. A corresponding protein synthesis, however, was obtained only with higher doses (10-100 ng/mL). The LPS-induced increase of mPGES-1 was inhibited by different signaling pathway inhibitors, such as SP600125, LY294002, GF109203X, and SC-514, suggesting the involvement of c-Jun N-terminal kinase (JNK), phosphatidylinositol 3-kinase (PI-3K)/Akt, protein kinase C (PKC) pathways, and the nuclear factor (NF)-kappaB, respectively. In contrast to other reports, LPS-induced mPGES-1 synthesis was not invariably coupled to the synthesis of COX-2, since inhibition of PI-3K with LY294002 decreased mPGES-1 but increased COX-2 levels. This detailed view of the intracellular signaling pathways involved in mPGES-1 expression in activated microglia opens a new avenue in the search for novel potential therapeutic targets to reduce neuroinflammation, and demonstrates that mPGES-1 expression is not strictly coupled to the expression of COX-2. PMID- 18383342 TI - HIV-1-infected and/or immune-activated macrophage-secreted TNF-alpha affects human fetal cortical neural progenitor cell proliferation and differentiation. AB - Neurogenesis, tied to the proliferation, migration and differentiation of neural progenitor cells (NPC) is affected during neurodegenerative diseases, but how neurogenesis is affected during HIV-1 associated dementia (HAD) has not been fully addressed. Here we test the hypothesis that HIV-1-infected and/or immune activated brain macrophages affect NPC proliferation and differentiation through the regulation of cytokines. We showed that human monocyte-derived macrophages (MDM) conditioned medium (MCM) induces a dose dependent increase in NPC proliferation. Conditioned media from lipopolysaccharide (LPS)-activated MDM (LPS MCM) or HIV-infected MCM (HIV-MCM) induced a profound increase in NPC proliferation. HIV-infected and LPS-activated MCM (HIV+LPS-MCM) induced the most robust increase in NPC proliferation. Moreover, LPS-MCM and HIV+LPS-MCM decreased beta-III-tubulin and increased GFAP expression, demonstrating an induction of gliogenesis and inhibition of neurogenesis. The increase of NPC proliferation and gliogenesis correlated with increases in production of TNF-alpha by infected/activated MDM. Although both IL-1beta and TNF-alpha induced NPC proliferation and gliogenesis, these effects were only partially abrogated by soluble TNF-alpha receptors R1 and R2 (TNF-R1R2), but not by the IL-1 receptor antagonist (IL-1ra). This indicated that the HIV-1-infected/LPS-activated MCM mediated effects were, in part, through TNF-alpha. These observations were confirmed in severe combined immunodeficient (SCID) mice with HIV-1 encephalitis (HIVE). In these HIVE mice, NPC injected with HIV-infected MDM showed more astrocyte differentiation and less neuronal differentiation compared to NPC injection alone. These observations demonstrated that HIV-1-infected and immune activated MDM could affect neurogenesis through induction of NPC proliferation, inhibition of neurogenesis, and activation of gliogenesis. PMID- 18383344 TI - Replication of Theiler's virus requires NF-kappa B-activation: higher viral replication and spreading in astrocytes from susceptible mice. AB - To investigate viral replication and cell-cell spreading in astrocytes, recombinant Theiler's murine encephalomyelitis virus (TMEV) expressing green fluorescent protein (GFP) during the replication was generated. GFP and TMEV proteins were processed correctly in infected cells and production of viral proteins could be tracked by fluorescent microscopy. Viral replication of both wild-type TMEV and GFP-TMEV was dependent on the activation of NF-kappaB and partially MAP kinase, based on chemical inhibition studies. Viral replication was significantly reduced in primary astrocytes from NF-kappaB1 (p105)-deficient mice compared with that from wild-type control mice, whereas cytokine production was enhanced. These results suggest an association of canonical NF-kappaB subunits in viral replication, but not cytokine production. Viral replication was also suppressed in both IKKalpha and IKKbeta-deficient mouse embryonic fibroblasts (MEFs), compared with that in wild-type MEF. However, the inhibition was significantly greater in IKKbeta-deficient MEF, suggesting that IKKbeta plays a stronger role in supporting viral replication. Interestingly, viral replication and spreading in primary astrocytes from susceptible SJL/J mice were several-fold higher than those in astrocytes from resistant C57BL/6 mice, suggesting that higher viral replication levels in astrocytes may also contribute to the viral persistence in the central nervous system (CNS) of susceptible SJL/J mice. A relatively higher level of activated NF-kappaB was found in the nuclei of virus infected SJL astrocytes compared with C57BL/6 astrocytes suggest that the NF kappaB activation level affects on viral replication. PMID- 18383343 TI - H-Ras increases urokinase expression and cell invasion in genetically modified human astrocytes through Ras/Raf/MEK signaling pathway. AB - Previous study reported that the activation of Ras pathway cooperated with E6/E7 mediated inactivation of p53/pRb to transform immortalized normal human astrocytes (NHA/hTERT) into intracranial tumors strongly resembling human astrocytomas. The mechanism of how H-Ras contributes to astrocytoma formation is unclear. Using genetically modified NHA cells (E6/E7/hTERT and E6/E7/hTERT/Ras cells) as models, we investigated the mechanism of Ras-induced tumorigenesis. The overexpression of constitutively active H-RasV12 in E6/E7/hTERT cells robustly increased the levels of urokinase plasminogen activator (uPA) mRNA, protein, activity and invasive capacity of the E6/E7/hTERT/Ras cells. However, the expressions of MMP-9 and MMP-2 did not significantly change in the E6/E7/hTERT and E6/E7/hTERT/Ras cells. Furthermore, E6/E7/hTERT/Ras cells also displayed higher level of uPA activity and were more invasive than E6/E7/hTERT cells in 3D culture, and formed an intracranial tumor mass in a NOD-SCID mouse model. uPA specific inhibitor (B428) and uPA neutralizing antibody decreased uPA activity and invasion in E6/E7/hTERT/Ras cells. uPA-deficient U-1242 glioblastoma cells were less invasive in vitro and exhibited reduced tumor growth and infiltration into normal brain in xenograft mouse model. Inhibitors of Ras (FTA), Raf (Bay 54 9085) and MEK (UO126), but not of phosphatidylinositol 3-kinase (PI3K) (LY294002) and of protein kinase C (BIM) pathways, inhibited uPA activity and cell invasion. Our results suggest that H-Ras increased uPA expression and activity via the Ras/Raf/MEK signaling pathway leading to enhanced cell invasion and this may contribute to increased invasive growth properties of astrocytomas. PMID- 18383345 TI - Statin treatment of adult human glial progenitors induces PPAR gamma-mediated oligodendrocytic differentiation. AB - The statins have been proposed as possible therapeutic agents for a variety of autoimmune disorders, including multiple sclerosis. In a genomic screen, we found that glial progenitor cells (GPCs) of the adult human white matter expressed significant levels of the principal statin target, HMG-CoA reductase, as well as additional downstream members of the sterol synthesis pathway. We therefore asked if statin treatment might influence the differentiated fate of adult glial progenitor cells. To assess the functional importance of the sterol synthesis pathway to adult human glial progenitors, we used simvastatin or pravastatin to inhibit HMG-CoA reductase, and then assessed the phenotypic differentiation of the progenitors, as well as the molecular concomitants thereof. We found that both statins induced a dose-dependent induction of oligodendrocyte phenotype, and concomitant reduction in progenitor number. Oligodendrocyte commitment was associated with induction of the sterol-regulated nuclear co-receptor PPARgamma, and could be blocked by the specific PPARgamma antagonist GW9662. Thus, statins may promote oligodendrocyte lineage commitment by parenchymal glial progenitor cells; this might reduce the available progenitor pool, and hence degrade the long-term regenerative competence of the adult white matter. PMID- 18383346 TI - Modulation of astrocytic metabolic phenotype by proinflammatory cytokines. AB - Astrocytes play an important role in nervous system homeostasis. In particular, they contribute to the regulation of local energy metabolism and to oxidative stress defence. In previous experiments, we showed that long-term treatment with interleukin 1alpha (IL-1alpha) or tumor necrosis factor-alpha (TNFalpha) alone increases glucose utilization in primary culture of mouse astrocytes. In our study, we report that a combination of IL-1beta and TNFalpha exerts a synergistic effect on glucose utilization and markedly modifies the metabolic phenotype of astrocytes. Thus, IL-1beta+TNFalpha treated astrocytes show a marked decrease in glycogen levels, a slight but not significant decrease in lactate release as well as a massive increase in both the pentose phosphate pathway and TCA cycle activities. Glutamate-stimulated glucose utilization and lactate release, a typical feature of astrocyte energy metabolism, are altered after pretreatment with IL-1beta+TNFalpha. As far as mechanisms for oxidative stress defence are concerned, we observed that treatment with IL-1beta+TNFalpha decreases cellular glutathione content and increases glutathione release into the extracellular space while stimulating superoxide anion and nitric oxide production as well as H(2)O(2) release. Interestingly, stimulation of glucose utilization by IL 1beta+TNFalpha is not affected by the antioxidant N-acetyl-L-cysteine, suggesting that cellular stress does not account for this effect. Finally, the effects of cytokines on glucose utilization appear to involve multiple signaling cascades including the phosphoinositide 3-kinase and mitogen-activated protein kinases. Taken together these results establish that a proinflammatory environment such as observed in several neuropathological conditions including Alzheimer's disease, markedly modifies the metabolic phenotype of astrocytes. PMID- 18383347 TI - Protective effects of glucagon-like peptide 2 on intestinal ischemia-reperfusion rats. AB - Our objective was to evaluate the protective effects of glucagon-like peptide 2 (GLP-2) on intestinal ischemia/reperfusion (I/R) rats. Thirty-two rats were randomly assigned to four experimental groups, each of 8: Group A, sham rats underwent laparotomy only, without superior mesenteric artery (SMA) occlusion; Group B, I/R animals underwent laparotomy and occlusion of the SMA for 60 minutes followed by 120 minutes of reperfusion; Group C, I/R animals underwent intestinal I/R, and received pretreatment with GLP-2 for 3 days preoperatively; and Group D, I/R animals underwent intestinal I/R, received pretreatment with GLP-2 as above, and during the reperfusion phase were injected intravenously with GLP-2. After the reperfusion of intestinal ischemia, samples of intestinal mucosa, mesenteric lymph nodes (MLN) and blood were prepared for determination. In the pretreatment rats with GLP-2 (group C), Chiu's scores, bacterial colony counts, serum D lactate, intestinal mucosal MDA and ET-1, and serum endotoxin, TNF-alpha and IL-6 were significantly reduced compared with intestinal I/R rats (group B). Administration of GLP-2 during the reperfusion phase following pretreatment (group D) showed further protective effects in comparison with the pretreatment rats (group C). We conclude that treatment with GLP-2 attenuates intestinal I/R injury, reduces bacterial translocation, inhibits the release of oxygen free radicals and ET-1, and may well inhibit the production of proinflammatory cytokines. PMID- 18383348 TI - Transplantation of a vascularized rabbit femoral diaphyseal segment: mechanical and histologic properties of a new living bone transplantation model. AB - A new vascularized bone transplantation model is described, including the anatomy and surgical technique of isolating a rabbit femoral diaphyseal segment on its nutrient vascular pedicle. The histologic and biomechanical parameters of pedicled vascularized femoral autotransplants were studied following orthotopic reimplantation in the resulting mid-diaphyseal defect. Vascularized femur segments were isolated in 10 rabbits on their nutrient pedicle, and then replaced orthotopically with appropriate internal fixation. Postoperative weightbearing and mobility were unrestricted, and the contralateral femora served as no treatment controls. After 16 weeks, the bone flaps were evaluated by x-ray (bone healing), mechanical testing (material properties), microangiography (quantification of intraosseous vasculature), histology (bone viability), and histomorphometry (bone remodeling). Bone healing occurred by 2 weeks, with further callus remodeling throughout the survival period. Eight transplants healed completely, while two had a distal pseudarthrosis. Microangiography demonstrated patent pedicles in all transplants. Intraosseous vessel densities were comparable to nonoperated (control) femora. We found ultimate strength and elastic modulus to be significantly reduced when compared to normal controls. Viable bone, increased mineral apposition rate, and bone turnover were demonstrated in all transplants. The method described, and the data provided will be of value for the further study of isolated segments of living bone, and in particular, for investigations of reconstruction of segmental bone loss in weight bearing animal models. This study also provides important normative data on living autologous bone flap material properties, vascularity, and bone remodeling. We intend to use this method and data for comparison in subsequent studies of large bone vascularized allotransplantation. PMID- 18383349 TI - Reliability of the superficial venous drainage of the radial forearm free flaps in oral and maxillofacial reconstruction. AB - The aim of this article is to compare the superficial venous drainage system versus dual venous drainage system of radial forearm free flaps (RFFF) and its impact on flap survival. A total of 136 consecutive patients undergoing RFFF reconstruction for oral and maxillofacial surgical defects were reviewed retrospectively. In group l, both the superficial and deep veins were anastomosed to the neck recipient veins. In group 2, only the superficial vein was used for anastomosis. There were no statistical differences in venous problems between two groups (6/68 in group 1 vs. 5/68 in group 2, P = 0.75). The flap survival rate was 98.5% (66/68 in group 1 vs. 68/68 in group 2, P = 0.15). Although there are many theoretical advantages to dual venous drainage system anastomoses, they seem not to impact on flap survival in our experience. PMID- 18383350 TI - Repairing a 35-mm-long median nerve defect with a chitosan/PGA artificial nerve graft in the human: a case study. AB - We have developed a chitosan/polyglycolic acid (PGA) artificial nerve graft which was previously used for bridge implantation of dog sciatic nerves across 30-mm long defects. Here we describe a clinical trial of this graft for repairing a 35 mm-long median nerve defect at elbow of a human patient. During the 3-year follow up period, functional recovery of the injured median nerve was assessed by pinch gauge test, hydraulic hand dynamometry, static two-point discrimination and touch test with monofilaments, in couple with electrophysiological examinations. The motor and sensory function of the median nerve demonstrated an ongoing recovery postimplantation, reaching M4 and S3+ levels during the follow-up period. The results indicate that the chitosan/PGA artificial nerve graft could be used for surgical repair of larger defects in major peripheral nerves at a higher level in the human. PMID- 18383351 TI - Thiel embalming technique: a valuable method for microvascular exercise and teaching of flap raising. AB - Because of its high requirements on dexterity and microsurgical skills and the need of complete understanding of flap anatomy, microvascular free flap transfer belongs to the most demanding surgical procedures. Therefore, courses for flap raising and microvascular exercise are considered a prerequisite to prepare for clinical practise. To achieve teaching conditions as realistic as possible we used a novel cadaver embalming method enabling tissue dissection comparable to the living body. Thirty cadavers which were offered to us by the Institute of Anatomy for the purpose of running flap raising courses were embalmed in the technique described by Thiel. On each cadaver, nine free flaps were dissected according to a structured protocol by each course participant and afterwards used for microvascular exercise. The conservation of fine vascular structures and the suitability of the embalmed tissue for microvascular suturing were observed and photographically documented. The Thiel embalming technique provided flap raising procedures to be performed under realistic conditions similar to the living body. Vessels and nerves could be exposed and dissected up to a diameter of 1 mm and allowed for microvascular suturing even after weeks like fresh specimens. The Thiel embalming method is a unique technique and ideally suited to teach flap raising and microvascular suturing on human material. PMID- 18383352 TI - Decreasing the expression of LFA-1 and ICAM-1 as the major mechanism for the protective effect of hyperbaric oxygen on ischemia-reperfusion injury. PMID- 18383353 TI - Simultaneous coverage of two separate dorsal digital defects with a syndactylizing venous free flap. AB - We report a case of a 58-year-old patient who sustained multiple finger injuries on the right hand. After thorough debridement, two dorsal defects of the middle and ring fingers were covered simultaneously with a single arterialized venous free flap from the right forearm. The flap was used to create a dorsally syndactylized digit which survived completely and was subsequently divided longitudinally. With early flap division, excellent functional and cosmetic result was achieved. We consider the syndactylized venous free flap with early division as a useful option for simultaneous coverage of separate dorsal digital defects. PMID- 18383354 TI - Functional evaluation of anterolateral thigh flap donor sites: isokinetic torque comparisons for knee function. AB - BACKGROUND: The anterolateral thigh flap is thought to have minor donor site morbidity, but muscle dissection is unavoidable when skin perforator vessels run through the vastus lateralis muscle. The purpose of this study was to investigate the functional problems associated with the anterolateral thigh flap donor site. METHOD: We evaluated 12 patients who underwent free anterolateral thigh flap transfer between March 2003 and November 2005. A questionnaire and dynamic functional evaluation of the knee joint using the Biodex System were performed preoperatively and 6 months postoperatively. RESULTS: No patients reported any disturbance in their daily life. No significant differences were found between donor and normal thighs on isokinetic power tests of the quadriceps muscle. CONCLUSION: The function of the donor site after harvesting the anterolateral thigh flap was maintained. Damage to or functional disturbance of the donor site is minimal even if muscle is injured when harvesting the flap. PMID- 18383355 TI - Comparative geometric analysis of diamond and hole techniques in end-to-side microvascular anastomosis. AB - Although microsurgical techniques have been accepted routinely in many plastic surgical operations, many experimental studies are currently being conducted to refine the results. Most of these studies are difficult to perform and evaluate and the results are not always free of bias. Many scientific disciplines recently use the mathematics for simulation and modeling to predict the results of difficult experiments. Slit and hole end-to-side microanastomosis techniques are frequently preferred by many microsurgeons. Diamond technique recently proposed by the senior author is also a technique of end-to-side anastomosis with some advantages. The authors used mathematical simulation and modeling to compare two end-to-side microanastomosis techniques namely hole and diamond. The authors measured cross sectional areas of anastomosis in both techniques with different input data. The results showed that for end-to-side anastomosis, at angles less than 60 degrees , diamond technique yields a greater cross sectional area at anastomosis site compared with that of hole technique. PMID- 18383356 TI - Biologic drugs for rheumatoid arthritis in the Medicare program: a cost effectiveness analysis. AB - OBJECTIVE: Since the introduction of the Medicare Prescription Drug Improvement and Modernization Act and its associated demonstration project, coverage of selected biologic drugs has been expanded for Medicare beneficiaries. For rheumatoid arthritis, coverage was extended to etanercept, adalimumab, and anakinra in addition to the previously covered infliximab. We undertook to develop a model to compare the costs and quality-adjusted life years (QALYs) generated by each of the 4 biologic agents. METHODS: Data were drawn from meta analysis of randomized controlled trials and from a large longitudinal outcomes databank. Uncertainty was addressed using probabilistic and one-way sensitivity analyses. A lifetime horizon and Medicare viewpoint were adopted. RESULTS: In the base case analysis, anakinra was the least effective and least costly strategy. Etanercept, adalimumab, and infliximab were similar in terms of effectiveness, but infliximab was more costly. If decision makers are willing to pay a maximum of $50,000/QALY, the probability that infliximab is cost-effective is <1%. Findings were robust to a range of sensitivity analyses. Only if the dose of infliximab remains constant over time is this likely to be a cost-effective strategy. CONCLUSION: Infliximab is unlikely to be cost-effective in the Medicare population compared with either etanercept or adalimumab. Anakinra is substantially less costly but is also less effective than the 3 tumor necrosis factor alpha inhibitors. PMID- 18383357 TI - Increased cortisol relative to adrenocorticotropic hormone predicts improvement during anti-tumor necrosis factor therapy in rheumatoid arthritis. AB - OBJECTIVE: Some patients with chronic inflammatory diseases such as rheumatoid arthritis (RA) improve rapidly from anti-tumor necrosis factor (anti-TNF) therapy. No sensitive markers are available that might predict outcome of anti TNF therapy. We undertook this study to investigate the predictive value of hypothalamic-pituitary-adrenal (HPA) axis hormones for clinical improvement during anti-TNF therapy. METHODS: An observational study in 23 RA patients was followed by a validation study in 38 RA patients. The patients receiving anti-TNF antibodies had no glucocorticoid treatment, and we measured baseline serum levels of adrenocorticotropic hormone (ACTH) and cortisol. Improvement during anti-TNF antibody treatment was judged by the Disease Activity Score in 28 joints (DAS28), and serum levels of cortisol were measured at followup. RESULTS: The observational study demonstrated that improvement in the DAS28 correlated negatively with baseline serum levels of cortisol (R=-0.520, P=0.011) and the cortisol:ACTH ratio (R=-0.700, P=0.0002). In the longitudinal part of the study at followup, those patients with good improvement and initially low serum levels of cortisol demonstrated an increase of serum cortisol, in contrast to patients with little or no improvement. Findings in the observational study were supported by those in the validation study in a group of RA patients with less inflammation (correlation of improvement in the DAS28 with cortisol:ACTH ratio: R=-0.320, P=0.025). CONCLUSION: This is the first study in a human chronic inflammatory disease to demonstrate that inflammation-induced TNF interferes with HPA axis integrity, which is linked to the disease outcome. These findings position the HPA axis centrally in the vicious circle of perpetuation of chronic inflammation. PMID- 18383359 TI - CD200-Fc, a novel antiarthritic biologic agent that targets proinflammatory cytokine expression in the joints of mice with collagen-induced arthritis. AB - OBJECTIVE: The CD200 receptor (CD200R) is an inhibitory receptor expressed by myeloid cells that is postulated to play an important role in regulation of the immune system. The purpose of this study was to evaluate the efficacy of a soluble ligand of CD200R in established collagen-induced arthritis (CIA) in mice and to analyze changes in cytokine expression following therapy in order to understand its primary mechanism of action. METHODS: Arthritis was induced in DBA/1 mice, and CD200-Fc fusion protein, an isotype control monoclonal antibody, or TNFR-Fc fusion protein was administered over a period of 10 days (total of 4 doses). Cytokine expression in the joint was assessed by flow cytometry, enzyme linked immunosorbent assay, and quantitative real-time polymerase chain reaction. RESULTS: CD200-Fc significantly reduced the severity of established arthritis at the clinical and histologic levels. The therapeutic effect of CD200-Fc at 1 mg/kg was comparable with that of TNFR-Fc at 4 mg/kg. CD200R was found to be expressed in arthritic synovia and in lymph nodes, yet no changes in T cell cytokine levels (interferon-gamma, interleukin-5 [IL-5], IL-10, IL-17) were detected after CD200 Fc therapy. There was no evidence of an expansion of forkhead box P3-positive regulatory T cells or a change in serum anticollagen IgG1 and IgG2a levels. However, administration of CD200-Fc markedly decreased the expression of messenger RNA for tumor necrosis factor alpha, IL-1beta, IL-10, and matrix metalloproteinase 13 in the joint to the same extent as administration of TNFR Fc. CONCLUSION: CD200-Fc is an effective therapeutic agent in established CIA that targets proinflammatory cytokine expression in the joint without any obvious systemic immunosuppressive effects. Our findings indicate that CD200-Fc has considerable potential as a novel therapeutic agent in rheumatoid arthritis in humans. PMID- 18383358 TI - The influence of age at symptom onset and length of followup on mortality in patients with recent-onset inflammatory polyarthritis. AB - OBJECTIVE: To investigate the influence of age at symptom onset and length of followup on mortality in patients with recent-onset inflammatory polyarthritis (IP), and to examine predictors of mortality in relation to disease duration. METHODS: From 1990 to 1994, patients with recent-onset IP were registered with the Norfolk Arthritis Register (NOAR) and followed up prospectively. Standardized mortality ratios (SMRs) were calculated for all-cause and cardiovascular disease (CVD) mortality and for those who were younger than age 55 years at disease onset and for the first 5 and 10 years of followup. Cox proportional hazards models were developed to assess predictors of early and later mortality. RESULTS: Of 1,098 patients, 224 (20%) had died by the end of 2004. All-cause and CVD mortality were increased in rheumatoid factor (RF)-positive patients and in this subgroup, CVD mortality was increased at both early and later followup (SMR 5 year followup 1.93 [95% confidence interval 1.08-3.19]; SMR 10-year followup 2.00 [95% confidence interval 1.37-2.80]). CVD mortality was highest in seropositive patients<55 years of age at disease onset (SMR 5.58 [95% confidence interval 2.24 11.50]). In multivariate models, age at onset, male sex, RF positivity, Health Assessment Questionnaire score>or=1.5, and nodules were predictors of early and later mortality. CONCLUSION: Patients with IP had higher rates of CVD mortality throughout the followup period studied, and this was highest in seropositive patients who were <55 years of age at symptom onset. This subgroup deserves particular attention in terms of disease and risk factor modification. Nodules were independent predictors of CVD mortality, suggesting that extraarticular/vascular inflammation identifies patients at particularly high CVD risk. PMID- 18383360 TI - The effect of glycosaminoglycan loss on chondrocyte viability: a study on porcine cartilage explants. AB - OBJECTIVE: Loss of glycosaminoglycan (GAG) is an early event in osteoarthritis. Recent findings showed increased cell death in arthritic cartilage and linkage with extracellular matrix degradation. The aim of this study was to analyze the direct effect of GAG loss on chondrocyte survival and cell death following mechanical injury. METHODS: In full-thickness cartilage explants from porcine knee joints, GAG was depleted by digestion with chondroitinase ABC. Explants were subjected to single-impact mechanical injury. Cell viability and the types of cell death were analyzed by Live/Dead cell assay, staining for active caspase 3, and sensitivity to caspase inhibitor. RESULTS: GAG depletion did not directly lead to increased cell death. In chondroitinase ABC-treated explants, but not in control explants, mechanical injury caused an immediate reduction in cell viability (from 84.6% to 71.0%); the reduction was prominent in the superficial zone. This immediate cell death was not inhibited by the pancaspase inhibitor Z VAD-FMK, suggesting cell necrosis. During subsequent culture, viability in these explants decreased further, to 50.5% on day 3. The second wave of cell death was reduced by the addition of Z-VAD-FMK in chondroitinase ABC-treated explants and was also associated with activation of caspase 3, suggesting apoptotic mechanisms of cell death. CONCLUSION: These results indicate that GAG loss alone does not directly lead to chondrocyte death. In response to mechanical injury, there is an immediate induction of necrotic cell death that is seen only in GAG-depleted explants and primarily in the superficial zone. During subsequent culture, cell death spreads via apoptotic mechanisms. PMID- 18383362 TI - Clinical trials in pediatric rheumatology: comment on the editorial by Lehman. PMID- 18383361 TI - T cell polarization identifies distinct clinical phenotypes in scleroderma lung disease. AB - OBJECTIVE: Lung involvement is the leading cause of morbidity and mortality in systemic sclerosis (SSc; scleroderma), and interstitial lung disease (ILD) is the most common pulmonary manifestation. An abnormal profibrotic Th2/Tc2-polarized T cell response is postulated to mediate tissue damage and fibrosis. The aim of this study was to investigate whether a polarized T cell phenotype in SSc is associated with lung disease or other clinical manifestations of SSc. METHODS: Circulating T cells were characterized by flow cytometry in 62 patients with SSc and 36 healthy control subjects, using antibodies against CD3, CD4, CD8, chemokine receptor CCR5 (Th1/Tc1-specific), and prostaglandin D2 receptor CRTH2 (Th2/Tc2-specific). The ratio between CCR5 and CRTH2 T cell frequencies was used to quantify type 1 (high-ratio) or type 2 (low-ratio) immune polarization. RESULTS: Patients with SSc exhibited lower CCR5/CRTH2 T cell ratios than those exhibited by control subjects (P<0.0001), indicating a Th2/Tc2-polarized phenotype. Markedly reduced CCR5/CRTH2 T cell ratios were observed in SSc patients with ILD compared with SSc patients without ILD (P<0.0001), particularly in patients with active ILD (P<0.0001) compared with those with stable lung function. Lower CCR5/CRTH2 ratios were strongly associated with a lower value for the percent predicted forced vital capacity (P<0.0001). In patients with an estimated right ventricular systolic pressure>35 mm Hg, suggestive of pulmonary vascular disease, a lower value for the percent predicted diffusing capacity (DLCO) was associated with higher CCR5/CRTH2 T cell ratios (Th1/Tc1) (P=0.009), while in those with right ventricular systolic pressure<35 mm Hg, a lower value for the percent predicted DLCO correlated with lower ratios (Th2/Tc2) (P<0.0001), as observed for ILD. CONCLUSION: T cell polarization in SSc is strongly associated with specific manifestations of lung disease. Measurement of T cell polarization may represent a valuable tool to monitor disease activity and predict clinical outcomes in SSc patients with lung disease. PMID- 18383363 TI - Association of interleukin-23 receptor variants with ankylosing spondylitis. AB - OBJECTIVE: Recent studies have shown that a nonsynonymous single-nucleotide polymorphism (SNP) (Arg381Gln; rs11209026) in the interleukin-23 receptor (IL 23R) gene on chromosome 1p31 is associated with Crohn's disease and psoriasis. Given the clinical and immunologic overlap between ankylosing spondylitis (AS) and these diseases, and the potential function of this candidate SNP, this study was undertaken to examine the association of IL-23R variants with AS in multiple Canadian populations. METHODS: We examined 3 cohorts of AS patients from established rheumatic disease centers in Canada. The majority of AS patients were Caucasians of northern European descent, and all patients satisfied the modified New York classification criteria for AS or for juvenile spondylarthritis. We examined 424 AS probands and 401 controls from Alberta, 251 AS probands and 122 controls from Toronto, and 121 AS probands and 219 controls from Newfoundland. Ten IL-23R SNPs were genotyped, 9 of which were incorporated in the haplotype analysis. Allele and haplotype associations were calculated using the WHAP software package. P values for haplotype associations were calculated using a permutation test. RESULTS: The primary SNP of interest in a previous study of inflammatory bowel disease (IBD) (Arg381Gln; rs11209026) was found to be protective against AS in the Newfoundland population (P=0.04) and in the Toronto population (P=0.04) in single-marker univariate analysis. The strongest association, however, was with SNP rs11465804 (P=0.007 for the Newfoundland population and P=0.0007 for the Toronto population). A 3-marker sliding window omnibus test revealed a significant association with markers rs10489629, rs2201841, and rs11465804 in both the Newfoundland population (P=0.04) and the Alberta population (P=0.034). Our results were independent of the IBD and psoriasis status of the AS patients. CONCLUSION: This concurrent analysis of 3 distinct AS populations and their regional controls demonstrates a disease association with the IL-23R locus and implicates the same polymorphisms associated with IBD and psoriasis. PMID- 18383364 TI - A type I collagen defect leads to rapidly progressive osteoarthritis in a mouse model. AB - OBJECTIVE: This study was undertaken to test the hypothesis that abnormalities of the subchondral bone can result in osteoarthritis (OA). METHODS: We used a knockin model of human osteogenesis imperfecta, the Brittle IV (Brtl) mouse, in which defective type I collagen is expressed in bone. OA in individual mice was documented by micro-magnetic resonance imaging (micro-MRI) and micro-computed tomography (micro-CT). Alterations in the knee joints were confirmed by histopathologic and immunohistochemical analysis. In addition, atomic force microscopy (AFM) was used to assess the ultrastructure of the articular cartilage and subchondral bone matrix. RESULTS: Brtl mice had decreased integrity of bone but initially normal articular cartilage. However, by the second month of life, Brtl mice developed alterations of the cartilage that were characteristic of OA, as documented by micro-CT, micro-MRI, and histologic evaluation. In addition, chondrocyte loss and breakdown of the collagen matrix in the residual cartilage were demonstrated using AFM. CONCLUSION: The Brtl mouse model demonstrates that progressive destruction of articular cartilage characteristic of OA may be secondary to altered architecture of the underlying subchondral bone. PMID- 18383365 TI - Sialic acid-binding Ig-like lectin 1 expression in inflammatory and resident monocytes is a potential biomarker for monitoring disease activity and success of therapy in systemic lupus erythematosus. AB - OBJECTIVE: Type I interferon (IFN) plays a pivotal role in the pathogenesis of systemic lupus erythematosus (SLE) and is therefore considered a potential therapeutic target. This study was undertaken to establish a feasible biomarker for IFN effects with respect to disease activity and effectiveness of IFN suppressive therapy in SLE patients. METHODS: Transcriptomes of purified monocytes from 9 SLE patients and 7 healthy controls were analyzed by Affymetrix GeneChip technology. Levels of sialic acid-binding Ig-like lectin 1 (Siglec-1) (sialoadhesin, CD169) in inflammatory and resident monocytes were determined at the protein level in 38 healthy controls and 52 SLE patients, using multicolor flow cytometry. RESULTS: Transcriptomes of peripheral monocytes from SLE patients revealed a dominant type I IFN signature. Siglec-1 was identified as one of the most prominent type I IFN-regulated candidate genes. At the protein level, the frequency of Siglec-1-expressing monocyte subsets was correlated with disease activity (as measured by the SLE Disease Activity Index) and was inversely correlated with levels of complement factors. Most interestingly, levels of anti double-stranded DNA (anti-dsDNA) antibodies were highly correlated with the percentage of resident monocytes, but not inflammatory monocytes, expressing Siglec-1. High-dose glucocorticoid treatment resulted in a dramatic reduction of Siglec-1 expression in cells from patients with active SLE. CONCLUSION: Our findings indicate that Siglec-1 expression in resident blood monocytes is a potential biomarker for monitoring disease activity, displaying type I IFN responses, and estimating levels of anti-dsDNA antibodies. Moreover, our results suggest that resident and inflammatory monocytes contribute differently to the process of autoantibody formation in SLE. PMID- 18383366 TI - Performing trials in children with rheumatic diseases: comment on the editorial by Lehman. PMID- 18383368 TI - Coexpression of two mesenchymal cadherins, cadherin 11 and N-cadherin, on murine fibroblast-like synoviocytes. AB - OBJECTIVE: Cadherin 11 has recently been identified on fibroblast-like synoviocytes (FLS), and studies in mice have demonstrated its importance in synovial lining architecture and inflammation. However, many tissues express more than 1 cadherin. Given the newly appreciated functional significance of cadherins in the synovium, this study was undertaken to determine whether mouse FLS express other cadherins in addition to cadherin 11. METHODS: The characterization of cadherin expression was determined in FLS derived from wild-type and cadherin 11 null mice using immunofluorescence (IF), biochemical, and immunohistologic techniques. RESULTS: Cadherin 11 expression was observed at points of cell-cell contact in cultured wild-type mouse FLS. However, despite the lack of cell surface cadherin 11, cadherin 11-null mouse FLS cells still formed intimate cell cell contacts that contained beta-catenin. Immunoprecipitation of cell surface biotinylated FLS with anti-beta-catenin antibody demonstrated the presence of 2 cell surface catenin-associated proteins in FLS from wild-type mice and 1 in FLS sample from cadherin 11-null mice. Using biochemical approaches and reverse transcriptase-polymerase chain reaction, these proteins were determined to be N cadherin and cadherin 11. Expression of both N-cadherin and cadherin 11 in the synovial lining was demonstrated by immunohistochemical analysis of mouse synovium. IF analyses demonstrated colocalization of N-cadherin and cadherin 11 to the same points of cell-cell contact. However, the inability to coimmunoprecipitate both cadherins using either anti-N-cadherin or anti-cadherin 11 antibodies suggests that these cadherins are not contained within the same molecular complexes. CONCLUSION: These findings demonstrate that FLS express both N-cadherin and cadherin 11, and suggest that these cadherins are not contained within the same molecular complex. Given their importance in modulating cellular behavior, understanding how these cadherins regulate FLS behavior individually and in concert will be critical to understanding synovial architecture and inflammation. PMID- 18383369 TI - Familial clustering of non-nuclear autoantibodies and C3 and C4 complement components in systemic lupus erythematosus. AB - OBJECTIVE: To determine whether key features of systemic lupus erythematosus (SLE), namely, production of non-nuclear antibodies (anti-C1q and anticardiolipin antibodies [aCL]) and depletion of complement components C3 and C4, aggregate in families. In addition, we examined relationships between anti-C1q and C3 and C4 levels. METHODS: The study cohort comprised 1,037 predominantly white (82%) nuclear families in which at least 1 member had SLE. Associations of antibody measurements between probands and their unaffected siblings were examined using parametric and nonparametric analyses, along with associations between unaffected siblings and their parents. The heritability of anti-C1q, C3, and C4 was estimated, and interdependencies between these factors were examined in a regression model accounting for the family structure of the data set. RESULTS: We demonstrated associations between siblings for anti-C1q (odds ratio [OR] 3.74, 95% confidence interval [95% CI] 2.65, 5.28) and IgG and IgM aCL (OR 4.08, 95% CI 1.83, 5.13 and OR 2.06, 95% CI 1.46, 2.91, respectively) and, for anti-C1q, association between unaffected parents and their unaffected offspring (OR 4.34, 95% CI 2.16, 8.72). We also demonstrated significant heritability of anti-C1q, C3, and C4 (approximately 45%). Anti-C1q was negatively associated with C3 and C4 in SLE probands but not in their healthy relatives. CONCLUSION: Non-nuclear antibodies and C3 and C4 cluster within the families of SLE probands, suggesting that specific autoantibody formation is partly genetically determined, even if the total genetic effect in unaffected relatives is insufficient to cause disease. Anti-C1q antibodies accelerate C3 and C4 depletion in patients with SLE but have no effect in the absence of disease. PMID- 18383370 TI - Beta2-glycoprotein I protects thrombin from inhibition by heparin cofactor II: potentiation of this effect in the presence of anti-beta2-glycoprotein I autoantibodies. AB - OBJECTIVE: Beta2-glycoprotein I (beta2GPI) is an important autoantigen in the antiphospholipid syndrome (APS). In vitro studies suggest that it may have multifaceted physiologic functions, since it displays both anticoagulant and procoagulant properties. We have previously reported that beta2GPI can directly bind thrombin, a key serine protease in the coagulation pathway. The present study was undertaken to examine the influence of beta2GPI on thrombin inactivation by the serpin heparin cofactor II (HCII). The effect of anti beta2GPI antibodies was also examined. METHODS: HCII inactivation of thrombin was assessed using chromogenic and various platelet functional assays. The influence of intact and proteolytically cleaved beta2GPI and anti-beta2GPI antibodies was determined in these systems. RESULTS: beta2GPI protected thrombin against inactivation by HCII/heparin. Cleavage of beta2GPI at Lys317-Thr318 abrogated its protective effect. Patient polyclonal IgG and murine monoclonal anti-beta2GPI antibodies potentiated the procoagulant influence of beta2GPI in this system. CONCLUSION: These novel findings suggest that beta2GPI may regulate thrombin inactivation by HCII/heparin. The observation that anti-beta2GPI antibodies potentiate the protective effect of beta2GPI on thrombin in this system, thereby promoting a procoagulant response, may potentially delineate one of the pathophysiologic mechanisms contributing to the prothrombotic tendency in patients with APS. PMID- 18383371 TI - Lack of evidence for erythema multiforme in skin reaction to adalimumab: comment on the article by Beuthien et al. PMID- 18383372 TI - In vivo delivery of lentiviral vectors expressing vasoactive intestinal peptide complementary DNA as gene therapy for collagen-induced arthritis. AB - OBJECTIVE: Vasoactive intestinal peptide (VIP) has been shown to exert potent immunomodulatory activity, and the use of lentiviral vectors has been found to be an effective means of gene delivery. The present study was therefore undertaken to investigate the feasibility and efficiency of gene therapy using lentiviral vectors expressing VIP (LentiVIP) for the treatment of rheumatoid arthritis (RA). METHODS: We evaluated the therapeutic potential of the gene therapy strategy in the collagen-induced arthritis (CIA) mouse model, administering the vectors at different phases of the disease. The inflammatory response was determined by measuring the levels of various inflammatory cytokines and chemokines in the joints and serum. The Th1-mediated response was evaluated by determining the proliferative response and cytokine profile of T cells stimulated with autoantigen. RESULTS: A single intraperitoneal injection of LentiVIP was highly effective in treating CIA. Mice with established, severe arthritis showed complete regression of the disease. The therapeutic effect of LentiVIP was associated with widespread biodistribution of the vector and increased VIP levels, especially in joints and lymphoid organs, and was mediated through a striking reduction of the 2 deleterious components of the disease, i.e., the autoimmune response (self-reactive Th1 cell activity and autoantibody production) and the inflammatory response. LentiVIP treatment also induced the generation and/or activation of CD4+,CD25+,FoxP3+ Treg cells in arthritic mice. CONCLUSION: Our findings show that in vivo administration of lentiviral vector expressing VIP produces one of the most potent therapeutic effects described so far in any animal model of RA. We propose that VIP gene transfer should be further investigated as a potential novel, effective treatment of RA and other chronic autoimmune disorders. PMID- 18383373 TI - A B cell apotope of Ro 60 in systemic lupus erythematosus. AB - OBJECTIVE: Previous studies have attempted to segregate anti-60-kd Ro/SSA (anti Ro 60) responses in systemic lupus erythematosus (SLE) and primary Sjogren's syndrome (SS) but have shown limited disease preference. The aim of the present study was to determine whether the presence of autoantibodies against an Ro 60 apotope (an epitope expressed on apoptotic cells) distinguishes anti-Ro 60 responses in SLE and primary SS. METHODS: Multiparameter flow cytometry was used to select early apoptotic cells and measure the simultaneous binding of annexin V, propidium iodide, and anti-Ro 60-positive IgG from SLE patients (n=21) and patients with primary SS (n=19). The specificity of the Ro 60 apotope was determined by inhibition experiments with recombinant and native Ro 60. RESULTS: Autoantibodies against the Ro 60 apotope were prevalent in SLE patients (13 of 21, 62%) and were rarely observed in patients with primary SS (1 of 19, 5%) (P=0.0002). Further, within SLE patients, autoantibodies to the Ro 60 apotope strongly distinguished patients with anti-Ro 60 alone (12 of 13, 92%) from those with both anti-Ro 60 and anti-La (1 of 8, 13%) (P=0.0005). When we considered all patients with anti-Ro 60 alone, the presence of autoantibodies to the Ro 60 apotope had both high sensitivity (92.3%) and high specificity (85.7%) for SLE compared with primary SS (P=0.0012). The presence of autoantibodies to the Ro 60 apotope may therefore be of diagnostic value in patients with isolated anti-Ro 60 responses. CONCLUSION: The preferential targeting of an Ro 60 apotope exposed on early apoptotic cells in a subset of SLE patients implies disease-specific pathways for the induction of anti-Ro 60 autoimmunity. PMID- 18383374 TI - Pulmonary capillary endothelial metabolic dysfunction: severity in pulmonary arterial hypertension related to connective tissue disease versus idiopathic pulmonary arterial hypertension. AB - OBJECTIVE: Pulmonary endothelial dysfunction is intertwined with the development and progression of pulmonary arterial hypertension (PAH). Pulmonary endothelium is an active metabolic tissue in healthy human subjects. This study was undertaken to determine the effects of PAH on pulmonary endothelial angiotensin converting enzyme (ACE) activity and to identify differences between common PAH types, i.e., PAH related to connective tissue disease (PAH-CTD) versus idiopathic PAH (IPAH). METHODS: Nineteen patients with PAH-CTD, 25 patients with IPAH, and 23 control subjects were evaluated. The single-pass transpulmonary percent metabolism (%M) and hydrolysis (both reflecting enzyme activity per capillary) of an ACE synthetic substrate were determined. In addition, the calculated functional capillary surface area (FCSA), normalized to body surface area (BSA), was determined. RESULTS: The %M values in patients with PAH-CTD (mean+/-SEM 53.6+/-3.6%) were significantly reduced compared with those in control subjects (P<0.01) and those in patients with IPAH (P<0.03), but were similar between the IPAH and control groups (mean+/-SEM 66.2+/-3.6% and 74.7+/-2.7%, respectively). Substrate hydrolysis was also significantly reduced in patients with PAH-CTD. The FCSA/BSA was significantly reduced in patients with PAH-CTD (mean+/-SEM 1,068+/ 118 ml/minute/m2) and in patients with IPAH (1,443+/-186 ml/minute/m2) compared with that in controls (2,948+/-245 ml/minute/m2; P<0.01 for both). At a given cardiac index, the FCSA/BSA tended to be lower in the PAH-CTD group than in the IPAH group. Moreover, unlike in IPAH, a linear relationship between the FCSA/BSA and the diffusing capacity for carbon monoxide (DLCO) was observed in PAH-CTD (r=0.54, P<0.03). CONCLUSION: The metabolically functional pulmonary capillary bed appears to be reduced to an equal extent in PAH-CTD and IPAH. However, %M and hydrolysis appear to be reduced in PAH-CTD but not in IPAH, reflecting relatively diminished ACE activity on the pulmonary capillary endothelial cells of patients with PAH-CTD, and showing that pulmonary endothelial metabolic function differs between PAH types. This study also provides the first functional evidence that a reduced DLCO value in patients with PAH-CTD is related to the degree of FCSA loss. PMID- 18383376 TI - MEK/ERK pathway inhibitors as a treatment for inflammatory arthritis might result in the development of lupus: comment on the article by Thiel et al. PMID- 18383377 TI - The risk of infection associated with tumor necrosis factor alpha antagonists: making sense of epidemiologic evidence. PMID- 18383378 TI - Benefits and risks of ankylosing spondylitis treatment with nonsteroidal antiinflammatory drugs. PMID- 18383379 TI - Genetic association of vasoactive intestinal peptide receptor with rheumatoid arthritis: altered expression and signal in immune cells. AB - OBJECTIVE: Vasoactive intestinal peptide (VIP) has been shown to be one of the endogenous factors involved in the maintenance of immune tolerance. Administration of VIP ameliorates clinical signs in various experimental autoimmune disorders. This study was undertaken to investigate whether the exacerbated inflammatory autoimmune response in rheumatoid arthritis (RA) might result directly from altered expression and/or signaling of VIP receptors in immune cells. METHODS: The effect of specific agonists of different VIP receptors on collagen-induced arthritis in mice was investigated by clinical and histologic assessment and measurement of cytokine and chemokine production. Expression of VIP receptor type 1 (VPAC1) in synovial cells and monocytes from RA patients was determined by flow cytometry. Potential associations of VPAC1 genetic polymorphisms with RA susceptibility were investigated. RESULTS: A VPAC1 agonist was very efficient in the treatment of experimental arthritis, and deficient expression of VPAC1 in immune cells of RA patients was associated with the predominant proinflammatory Th1 milieu found in this disease. Immune cells derived from RA patients were less responsive to VIP signaling than were cells from healthy individuals and showed reduced VIP-mediated immunosuppressive activity, rendering leukocytes and synovial cells more proinflammatory in RA. A significant association between multiple-marker haplotypes of VPAC1 and susceptibility to RA was found, suggesting that the reduced VPAC1 expression in RA-derived immune cells is associated with the described VPAC1 genetic polymorphism. CONCLUSION: These findings are highly relevant to the understanding of RA pathogenesis. They suggest that VIP signaling through VPAC1 is critical to maintaining immune tolerance in RA. In addition, the results indicate that VPAC1 may be a novel therapeutic target in RA. PMID- 18383380 TI - Modulation of aggrecan and ADAMTS expression in ovine tendinopathy induced by altered strain. AB - OBJECTIVE: To evaluate histologic, immunohistochemical, and molecular changes in tendon induced by altered strain in a large-animal model. METHODS: A full thickness partial-width laceration of the infraspinatus tendon was created in 5 sheep, while 5 sham-operated sheep were used as controls. Sheep were killed after 4 weeks, and 4 differentially stressed tendon regions (tensile or near bone attachment from overstressed or stress-deprived halves) were evaluated for histopathology, proteoglycan (PG) accumulation, and characterization of glycosaminoglycans and aggrecan catabolites. Gene expression of matrix components, enzymes, and inhibitors was analyzed by reverse transcriptase polymerase chain reaction. RESULTS: Histopathologic changes were detected in both overstressed and stress-deprived tensile tendon, but only in stress-deprived tendon near bone. In overstressed and stress-deprived tensile tendon, levels of keratan sulfate, chondroitin 4-sulfate, and chondroitin 6-sulfate were increased. In overstressed tensile tendon, levels of ADAMTS-generated aggrecan catabolites were increased. There was increased matrix metalloproteinase 13 (MMP-13) and decreased fibromodulin and decorin expression in all regions. Increased MMP-1, MMP-9, MMP-14, and ADAMTS-1 expression, and decreased type II collagen expression were restricted to stress-deprived tendon. In stress-deprived bone-attachment regions, messenger RNA (mRNA) for aggrecan was decreased, and ADAMTS was increased. In overstressed tensile tendon, aggrecan mRNA was increased, and ADAMTS was decreased. CONCLUSION: The distinct molecular changes in adjacent tissue implicate altered strain rather than humoral factors in controlling abnormal tenocyte metabolism, and highlight the importance of regional sampling. Tendon abnormalities induced by increased strain are accompanied by increased aggrecan, decreased ADAMTS, and low PG expression, which may negatively impact the structural integrity of the tissue and predispose to rupture. PMID- 18383381 TI - Articular cartilage repair using dedifferentiated articular chondrocytes and bone morphogenetic protein 4 in a rabbit model of articular cartilage defects. AB - OBJECTIVE: To observe redifferentiation of dedifferentiated chondrocytes after transplantation into the joint, and to evaluate the ability of dedifferentiated chondrocytes transduced with adenovirus containing bone morphogenetic protein 4 (BMP-4) to redifferentiate in vitro and in vivo in a rabbit model of articular cartilage defects. METHODS: Monolayer and pellet culture systems were used to evaluate the redifferentiation of dedifferentiated chondrocytes transduced with BMP-4. A rabbit model of partial-thickness articular cartilage defects was used to evaluate cartilage repair macroscopically and histologically, 6 and 12 weeks after transplantation with first-passage, fifth-passage, or transduced fifth passage chondrocytes. Histologic grading of the repaired tissue was performed. Expression of BMP-4 and the ability of transplanted cells to recover a chondrocytic phenotype were also assessed. RESULTS: BMP-4--expressing dedifferentiated chondrocytes recovered a chondrocytic phenotype in vitro. After transplantation into the joint, some of the dedifferentiated chondrocytes in the defect sites could undergo redifferentiation and formed matrix that displayed positive toluidine blue staining for glycosaminoglycans. Histologic scores of the regenerative tissue revealed significantly better cartilage repair in rabbits transplanted with BMP-4--expressing cells than in the other treatment groups. Staining with toluidine blue revealed expression of BMP-4 in the cells and in the matrix surrounding the cells. CONCLUSION: Some dedifferentiated chondrocytes can redifferentiate after transplantation into the load-bearing joint. BMP-4 can be used to induce redifferentiation of dedifferentiated chondrocytes in vitro and in vivo, which could help enhance articular cartilage repair. PMID- 18383382 TI - Clinical images: a patient with a tumor of the foot. PMID- 18383383 TI - Differential expression of vasoactive intestinal peptide and its functional receptors in human osteoarthritic and rheumatoid synovial fibroblasts. AB - OBJECTIVE: Vasoactive intestinal peptide (VIP) has shown potent antiinflammatory effects in murine arthritis and ex vivo in human rheumatoid arthritis (RA) synovial cells. To investigate the potential endogenous participation of this system in the pathogenesis of RA, we analyzed the expression and regulation of VIP and its functional receptors in human fibroblast-like synoviocytes (FLS) from patients with osteoarthritis (OA) and patients with RA. METHODS: The expression of VIP was studied by reverse transcription-polymerase chain reaction (RT-PCR), enzyme immunoassay, and immunofluorescence in cultured FLS, and by immunohistochemical analysis in synovial tissue. The expression and function of the potential VIP receptors in FLS were studied by RT-PCR, determination of intracellular cAMP production, cell membrane adenylate cyclase (AC) activity, and interleukin-6, CCL2, and CXCL8 production in response to VIP or specific agonists and antagonists. RESULTS: VIP expression was detected in human FLS at the messenger RNA and protein levels, and it was significantly decreased in RA FLS compared with OA FLS. VIP receptor type 1 (VPAC1) was the dominant AC-coupled receptor in OA FLS, in contrast with RA FLS, in which VPAC2 was dominant. Tumor necrosis factor alpha-treated OA FLS reproduced the VIP and VPAC receptor expression pattern of RA FLS. The antagonistic effects of VIP on FLS proinflammatory factor production were reproduced by VPAC1- and VPAC2-specific agonists in OA FLS and RA FLS, respectively. CONCLUSION: VIP expression is down regulated in RA and in tumor necrosis factor alpha-treated FLS, suggesting that down-regulation of this endogenous antiinflammatory factor may contribute to the pathogenesis of RA. In RA FLS, VPAC2 mediates the antiinflammatory effects of VIP, suggesting that VPAC2 agonists may be an alternative to VIP as antiinflammatory agents. PMID- 18383384 TI - Requirement of Toll-like receptor 7 for pristane-induced production of autoantibodies and development of murine lupus nephritis. AB - OBJECTIVE: The detection of high titers of antibodies against small nuclear ribonucleoproteins (snRNP) is a diagnostic finding in patients in whom systemic lupus erythematosus (SLE) is suspected. Endogenous RNA molecules within snRNP trigger Toll-like receptor 7 (TLR-7) activation in B cells and dendritic cells, leading to anti-snRNP antibody production, which is associated with the development of immune complex nephritis in SLE. The purpose of this study was to investigate the role of TLR-7 in anti-snRNP antibody production and renal disease in SLE induced by an exogenous factor in the absence of genetic predisposition, using the pristane-induced murine lupus model. METHODS: Serum autoantibodies, IgG isotypes, and cytokine levels in pristane-treated wild-type and TLR-7-deficient mice were analyzed by enzyme-linked immunosorbent assay. Histopathologic changes in mouse kidneys were determined by light immunofluorescence microscopy. Cell subsets in splenocytes and peritoneal lavage cells from the mice were examined by flow cytometry. RESULTS: We found that anti-snRNP antibody production induced by pristane treatment was entirely dependent on the expression of TLR-7, whereas anti-double-stranded DNA antibody production was not affected by a lack of TLR-7. Impaired anti-snRNP antibody production in TLR-7-deficient mice was paralleled by lower levels of glomerular IgG and complement deposits, as well as less severe glomerulonephritis. CONCLUSION: TLR-7 is specifically required for the production of RNA-reactive autoantibodies and the development of glomerulonephritis in pristane-induced murine lupus, a model of environmentally triggered SLE in the absence of genetic susceptibility to autoimmunity. Specific interference with TLR 7 activation by endogenous TLR-7 ligands may therefore be a promising novel strategy for the treatment of SLE. PMID- 18383387 TI - Identity of single-nucleotide polymorphisms used in a clinical pharmacogenetic model to predict the efficacy of methotrexate monotherapy: comment on the article by Wessels et al. PMID- 18383385 TI - Fibroblast-specific perturbation of transforming growth factor beta signaling provides insight into potential pathogenic mechanisms of scleroderma-associated lung fibrosis: exaggerated response to alveolar epithelial injury in a novel mouse model. AB - OBJECTIVE: To explore increased susceptibility to fibrosis following experimental injury to alveolar epithelial cells (AECs) in a novel transgenic mouse model of scleroderma with fibroblast-specific perturbation of transforming growth factor beta (TGFbeta) signaling (TbetaRIIDeltak-fib mice). METHODS: Wild-type (WT) and transgenic mice were injured with intratracheally administered saline or bleomycin, and the lungs were harvested for biochemical, histologic, and electron microscopic analysis. RESULTS: Electron microscopy revealed AEC abnormalities in the lungs of untreated transgenic mice and bleomycin-treated WT mice; the lungs of transgenic mice treated with bleomycin showed severe epithelial damage. Compared with lungs from bleomycin-treated WT mice, lungs from bleomycin-treated transgenic mice demonstrated increased fibroproliferation, myofibroblast persistence, and impaired hyperplasia and increased apoptosis of type II AECs. The lungs from saline-treated transgenic mice and those from bleomycin-treated WT mice had phenotypic similarities, suggesting enhanced susceptibility to minor epithelial injury in the transgenic strain. The level of collagen was increased in the lungs from transgenic mice compared with that in the lungs from WT mice after treatment with either bleomycin or saline. Persistent fibrosis in bleomycin treated transgenic mice was independent of ongoing neutrophil inflammation but was associated with impaired alveolar epithelial repair. CONCLUSION: These results suggest that in the context of fibroblast-specific perturbation of TGFbeta signaling, even minor epithelial injury induces significant fibrosis. The model supports a central role for TGFbeta in determining fibrosis and demonstrates that lung fibroblasts may regulate the response of AECs to injury. Our findings provide insight into likely pathogenic mechanisms in scleroderma associated pulmonary fibrosis. PMID- 18383389 TI - Mathematical modeling of the cause of tuberculosis during tumor necrosis factor blockade. AB - OBJECTIVE: Tumor necrosis factor (TNF) blockade increases the risk of tuberculosis (TB). The purpose of this study was to use Markov modeling to examine the contributions of reactivation of latent tuberculous infection (LTBI) and the progression of new infection with Mycobacterium tuberculosis to active TB due to TNF blockade. These 2 pathogenic mechanisms cannot otherwise be readily distinguished. METHODS: Monte Carlo simulation was used to represent the range of reported values for the incidence of TB associated with infliximab (TNF monoclonal antibody) and etanercept (soluble TNF receptor) therapy. Iterative methods were then used to identify for each pair of incidence rates the Markov model parameters that most accurately represented the distribution of time to onset of TB as reported to the Food and Drug Administration. RESULTS: Modeling revealed an apparent median monthly rate of reactivation of LTBI by infliximab treatment of 20.8%, which was 12.1 times that with etanercept treatment (P<0.001). In contrast, both drugs appeared to pose a high risk of progression of new M tuberculosis infection to active TB. Progression of new infection appeared to cause nearly half of the etanercept-associated cases; it became the predominant cause of infliximab-associated cases only after the first year. CONCLUSION: Despite sharing a common therapeutic target, infliximab and etanercept differ markedly in the rates at which they reactivate LTBI. Confirmation of these findings will require the application of molecular epidemiologic tools to studies of TB in future biologics registries. Hidden Markov modeling and Monte Carlo simulation are powerful tools for revealing otherwise hidden aspects of the pathogenesis of TB. PMID- 18383390 TI - Results of a two-year followup study of patients with rheumatoid arthritis who received a combination of abatacept and methotrexate. AB - OBJECTIVE: To evaluate the efficacy, radiographic changes, and safety of abatacept and methotrexate therapy through 2 years in a long-term extension of a previously published 1-year study. METHODS: Patients who received placebo during year 1 were switched to abatacept. Patients taking abatacept continued to take it. Efficacy and safety were assessed through 2 years. RESULTS: Of 539 patients enrolled in the initial 1-year study, 488 completed 1 year of the long-term extension (2% discontinued for lack of efficacy). At 2 years, patients taking abatacept had maintained their responses on the American College of Rheumatology (ACR) improvement criteria and the Disease Activity Score in 28 joints (DAS28; using the C-reactive protein [CRP] level), as well as their physical function (according to the Health Assessment Questionnaire [HAQ] disability index [DI]) and health-related quality of life (HRQOL; assessed with the Short Form 36 [SF 36] health survey), that were observed at the end of the double-blind period (year 1 versus year 2 values were 81.9% versus 80.3% for ACR 20% improvement, 25.4% versus 30.9% for a DAS28 [CRP] of <2.6, 71.8% versus 66.8% for the HAQ DI, and 9.7 versus 10.6 and 7.3 versus 7.2, respectively, for the mean change in the physical and mental components summary scores of the SF-36). In the abatacept group, post hoc analysis demonstrated further inhibition of radiographic progression during year 2 (57% reduction in mean change of total score in year 2 versus year 1; P<0.0001), and minimal radiographic progression was observed (mean change in total score from baseline was 1.1 and 1.6 at year 1 and 2, respectively). Rates of adverse events (AEs) and severe AEs were consistent throughout the cumulative period. CONCLUSION: The improvements in signs and symptoms, physical function, and HRQOL observed after 1 year of abatacept treatment were maintained through 2 years of treatment. This durability was accompanied by a safety profile consistent with that in the double-blind portion of the study. Radiographic progression was further inhibited in year 2 compared with year 1, suggesting an increasing effect of abatacept on the inhibition of structural damage in year 2. PMID- 18383391 TI - Defective proliferative capacity and accelerated telomeric loss of hematopoietic progenitor cells in rheumatoid arthritis. AB - OBJECTIVE: In rheumatoid arthritis (RA), telomeres of lymphoid and myeloid cells are age-inappropriately shortened, suggesting excessive turnover of hematopoietic precursor cells (HPCs). The purpose of this study was to examine the functional competence (proliferative capacity, maintenance of telomeric reserve) of CD34+ HPCs in RA. METHODS: Frequencies of peripheral blood CD34+,CD45+ HPCs from 63 rheumatoid factor-positive RA patients and 48 controls matched for age, sex, and ethnicity were measured by flow cytometry. Proliferative burst, cell cycle dynamics, and induction of lineage-restricted receptors were tested in purified CD34+ HPCs after stimulation with early hematopoietins. Telomere sequences were quantified by real-time polymerase chain reaction. HPC functions were correlated with the duration, activity, and severity of RA as well as its treatment. RESULTS: In healthy donors, CD34+ HPCs accounted for 0.05% of nucleated cells; their numbers were strictly age dependent and declined at a rate of 1.3% per year. In RA patients, CD34+ HPC frequencies were age-independently reduced to 0.03%. Upon growth factor stimulation, control HPCs passed through 5 replication cycles over 4 days. In contrast, RA-derived HPCs completed only 3 generations. Telomeres of RA CD34+ HPCs were age-inappropriately shortened by 1,600 bp. All HPC defects were independent of disease duration, disease activity, and smoking status, and were present to the same degree in untreated patients. CONCLUSION: In RA, circulating bone marrow-derived progenitor cells were diminished, and concentrations stagnated at levels typical of those in old control subjects. HPCs from RA patients displayed growth factor nonresponsiveness and sluggish cell cycle progression; marked telomere shortening indicated proliferative stress induced senescence. Defective HPC function independent of disease activity markers suggests bone marrow failure as a potential pathogenic factor in RA. PMID- 18383392 TI - Altered expression of MicroRNA in synovial fibroblasts and synovial tissue in rheumatoid arthritis. AB - OBJECTIVE: MicroRNAs (miRNA) have recently emerged as a new class of modulators of gene expression. In this study we investigated the expression, regulation, and function of miR-155 and miR-146a in rheumatoid arthritis (RA) synovial fibroblasts (RASFs) and RA synovial tissue. METHODS: Locked nucleic acid microarray was used to screen for differentially expressed miRNA in RASFs treated with tumor necrosis factor alpha (TNFalpha). TaqMan-based real-time polymerase chain reaction was applied to measure the levels of miR-155 and miR-146a. Enforced overexpression of miR-155 was used to investigate the function of miR 155 in RASFs. RESULTS: Microarray analysis of miRNA expressed in RASFs treated with TNFalpha revealed a prominent up-regulation of miR-155. Constitutive expression of both miR-155 and miR-146a was higher in RASFs than in those from patients with osteoarthritis (OA), and expression of miR-155 could be further induced by TNFalpha, interleukin-1beta, lipopolysaccharide, poly(I-C), and bacterial lipoprotein. The expression of miR-155 in RA synovial tissue was higher than in OA synovial tissue. Enforced expression of miR-155 in RASFs was found to repress the levels of matrix metalloproteinase 3 (MMP-3) and reduce the induction of MMPs 3 and 1 by Toll-like receptor ligands and cytokines. Moreover, compared with monocytes from RA peripheral blood, RA synovial fluid monocytes displayed higher levels of miR-155. CONCLUSION: This study provides the first description of increased expression of miRNA miR-155 and miR-146a in RA. Based on these findings, we postulate that the inflammatory milieu may alter miRNA expression profiles in resident cells of the rheumatoid joints. Considering the repressive effect of miR-155 on the expression of MMPs 3 and 1 in RASFs, we hypothesize that miR-155 may be involved in modulation of the destructive properties of RASFs. PMID- 18383393 TI - Association of cerebrospinal fluid anti-NR2 glutamate receptor antibodies with diffuse neuropsychiatric systemic lupus erythematosus. AB - OBJECTIVE: To explore the association of antibodies directed against N-methyl-D aspartate receptor subunit NR2 (anti-NR2) in cerebrospinal fluid (CSF) with neuropsychiatric manifestations in systemic lupus erythematosus (NPSLE). METHODS: Paired serum and CSF specimens were obtained from 56 patients with NPSLE (38 with diffuse psychiatric/neuropsychological syndromes [diffuse NPSLE] and 18 with neurologic syndromes or peripheral neuropathy [focal NPSLE]) and from 20 control patients with noninflammatory neurologic diseases. IgG anti-NR2 antibodies were measured by enzyme-linked immunosorbent assay using synthetic peptide containing the extracellular ligand-binding domain of NR2. The binding of affinity-purified anti-NR2 to human neuroblastoma cell line SK-N-MC was examined by flow cytometry. RESULTS: Purified anti-NR2 bound to the surface of SK-N-MC cells. Levels of anti NR2 antibodies in CSF were significantly elevated in patients with diffuse NPSLE compared with levels in control patients or those with focal NPSLE, whereas there were no significant differences in serum anti-NR2 levels among the 3 groups. In 31 of the 38 patients with diffuse NPSLE (81.6%) and 8 of the 18 patients with focal NPSLE (44.4%), CSF anti-NR2 levels were more than 3 SD above the mean level in the control patients (P=0.0120 by chi-square test). CONCLUSION: These results indicate that anti-NR2 is a constituent of antineuronal antibodies and, more importantly, that anti-NR2 antibodies in CSF, but not in serum, are associated with diffuse NPSLE. PMID- 18383394 TI - High prevalence of temporomandibular joint arthritis at disease onset in children with juvenile idiopathic arthritis, as detected by magnetic resonance imaging but not by ultrasound. AB - OBJECTIVE: To determine the prevalence of temporomandibular joint (TMJ) disease in a cohort of children with new-onset juvenile idiopathic arthritis (JIA), and to compare magnetic resonance imaging (MRI) with ultrasound (US) for the detection of acute and chronic changes of TMJ arthritis. METHODS: Between January 2005 and April 2007, children with newly diagnosed JIA were prospectively evaluated for TMJ arthritis. Prior to imaging, jaw pain and disability were assessed with questionnaires and physical examination. The TMJs of all patients were imaged with MRI and US within 8 weeks of diagnosis. RESULTS: Of the 32 patients enrolled, 78% were female, and the median age was 8.6 years (range 1.5 17.2 years). Acute TMJ arthritis was diagnosed in 75% of the children by MRI and in none by US; chronic arthritis was diagnosed in 69% by MRI and in 28% by US. Findings of both acute and chronic TMJ disease were detected by MRI in 53% of the patients. Of those with acute TMJ arthritis, 71% were asymptomatic, and 63% had normal findings on jaw examination. Fifty-six percent of patients with acute disease had an improved maximal incisal opening after corticosteroid injection. Among these responders, 56% had been asymptomatic and had normal jaw examination findings. CONCLUSION: TMJ arthritis was present in the majority of patients with new-onset JIA. Findings on MRI along with responses to treatment among asymptomatic patients with normal jaw examination findings suggest that a history review and physical examination are not sufficient to screen for TMJ disease. Our results also suggest that MRI and US findings are not well correlated, and that MRI is preferable for the detection of TMJ disease in new-onset JIA. PMID- 18383395 TI - Successful treatment of a patient with Takayasu arteritis using a humanized anti interleukin-6 receptor antibody. AB - Takayasu arteritis (TA) is a chronic inflammatory disease that involves the aorta and its major branches. Since overproduction of interleukin-6 (IL-6) seems to play a pathogenic role in TA, we used the anti-IL-6 receptor (IL-6R) antibody tocilizumab to treat a 20-year-old woman with refractory active TA complicated by ulcerative colitis (UC). Treatment with tocilizumab improved the clinical manifestations of TA and the abnormal laboratory findings in this patient and ameliorated the activity of UC. These results indicate that IL-6R inhibition with tocilizumab might be a future treatment option for TA. PMID- 18383397 TI - Inspire before you retire. PMID- 18383398 TI - Prescribing pattern of drugs in the treatment of osteoarthritis in Italian general practice: the effect of rofecoxib withdrawal. AB - OBJECTIVE: In October 2004, rofecoxib was removed from the world market because of an increased risk of myocardial infarction. The aim of the present study was to compare the trend of nonsteroidal antiinflammatory drug (NSAID) use and other analgesics in osteoarthritis (OA) treatment before and after rofecoxib withdrawal in Italian general practice. METHODS: From the Caserta-1 Local Health Service database, 97 general practitioners were recruited. Prevalence and incidence of use of any study drug were calculated within 1 year before and after rofecoxib withdrawal. RESULTS: One-year prevalence of nonselective and preferential NSAID use did not change after rofecoxib withdrawal, whereas coxib use fell from 4.4% (95% confidence interval [95% CI] 4.2-4.5%) in the period before rofecoxib withdrawal (period I) to 1.6% (95% CI 1.5-1.7%) in the period after withdrawal (period II). Weak opioids were used in no more than 0.4% (95% CI 0.3-0.5%) in period II, after their introduction to reimbursement in December 2004. Also, 1 year incidence of coxib decreased from 31.3 per 1,000 (95% CI 30.2-32.4%) in period I to 8.7 per 1,000 (95% CI 8.1-9.2%) in period II. The disappearance of rofecoxib was associated with replacement drugs such as newly marketed dexibuprofen and aceclofenac, whereas nimesulide use coincidentally decreased. CONCLUSION: Rofecoxib withdrawal has markedly changed the prescribing pattern of drugs that are used in OA-related pain treatment, with a striking decrease of coxib use in Italian general practice. Education strategies addressed to health professionals should be planned to improve the management of pain treatment, particularly in degenerative joint diseases. PMID- 18383399 TI - Prepared patients: internet information seeking by new rheumatology patients. AB - OBJECTIVE: To investigate to what extent and why new rheumatology patients access medical information online prior to first appointments and secondarily to ask whether they discuss information gained from the Internet with physicians. METHODS: Research was conducted in a teaching rheumatology clinic with a nonrandom sample of 120 English-speaking adults presenting for first appointments in rheumatology. Quantitative and qualitative data were gained in pre- and postappointment patient surveys and interviews, including online information gathering prior to first appointment, demographics, health status, information usage in patient-physician interactions, and satisfaction. Data were analyzed for significant relationships across variables and for qualitative insights into quantitative outcome measures. RESULTS: Of all patients, 87.5% looked up their symptoms or suspected condition prior to their first appointment and 62.5% of all patients sought that information on the Internet. Only 20% of online information seekers discussed that information with their physicians. Age and sex were significant predictors of Internet information seeking. Physician and patient appointment satisfaction was significantly higher when Internet information was discussed; however, most patients did not discuss their information seeking because they primarily feared being perceived as challenging their physician. CONCLUSION: The majority of patients research their conditions online prior to initial appointments, but are unlikely to discuss that research with physicians even though discussion is related to higher satisfaction. Physicians may want to consider strategies for enabling communication about online research. PMID- 18383400 TI - Comparison of the health assessment questionnaire disability index and the short form 36 physical functioning subscale using Rasch analysis: comment on the article by Taylor and McPherson. PMID- 18383401 TI - Early aggressive care and symptomatic recovery from whiplash: comment on the article by Cote et al. PMID- 18383402 TI - Introduction to special section: cost and social and psychological impact of rheumatic diseases. PMID- 18383403 TI - A pilot study of subclinical coronary atherosclerosis in systemic sclerosis: coronary artery calcification in cases and controls. AB - OBJECTIVE: Systemic sclerosis (SSc) is associated with vasculopathy and endothelial cell injury, which could potentially increase the risk of coronary atherosclerosis. Multidetector computed tomography, a noninvasive procedure, generates a coronary calcium score (CCS) as a marker for coronary atherosclerosis. Serum proinflammatory high-density lipoprotein (piHDL) is a potential novel marker of atherosclerotic risk. The objective of the pilot study was to determine 1) the prevalence of subclinical coronary atherosclerosis in SSc and 2) serum piHDL levels as a potential novel marker of atherosclerotic risk in SSc. METHODS: A cross-sectional study of 17 patients with SSc and 17 age-, sex-, and race-matched healthy controls in Cincinnati, Ohio, was conducted. Measurements included CCS; body mass index; lipid profile; and serum levels of high-sensitivity C-reactive protein, homocysteine, and piHDL. RESULTS: Patients with SSc were slightly older (mean 52.8 years) than control subjects (mean 50.6 years; P = 0.01). Coronary calcium was found in 12 participants (9 with SSc, 3 controls; P = 0.03). The mean +/- SD CCS in patients with SSc was significantly greater than the controls (126.6 +/- 251.0 versus 14.7 +/- 52.2; P = 0.003). Five patients with SSc (29%), but no controls, had detectable levels of piHDL (P = 0.06). CONCLUSION: Prevalence of subclinical coronary atherosclerosis is greater in patients with SSc compared with healthy controls. These findings should be confirmed in a larger study. PMID- 18383405 TI - The cutaneous lupus disease activity and severity index as a validated outcome measure for cutaneous lupus erythematosus: comment on the article by Stamm et al. PMID- 18383406 TI - Disability in valued life activities among individuals with systemic lupus erythematosus. AB - OBJECTIVE: To identify the prevalence of disability in a wide range of valued life activities (VLAs) among individuals with systemic lupus erythematosus (SLE), 1-year changes in such disability, and predictors of and changes in VLA disability. METHODS: Data were from 2 waves of a cohort of 829 individuals with SLE interviewed annually by telephone. VLA disability was assessed using a scale rating the difficulty of performing 21 activities. Scores were also calculated for subscales corresponding to obligatory, committed, and discretionary activities. Changes in VLA disability from baseline to 1-year followup were assessed. Sociodemographic and disease status measures were examined as predictors of and changes in VLA disability using multiple regression analyses. RESULTS: Almost half of the subjects were unable to perform > or =1 VLA at baseline. Almost all (91%) reported > or =1 VLA affected by SLE. One-quarter of the subjects experienced a significant increase in the number of activities they were unable to perform; approximately half experienced significant increases in the number of activities affected and in difficulty scores. Proportions of individuals whose disability increased and whose disability decreased were roughly equivalent. Disease status measures accounted for 62-72% of the variation in VLA difficulty. More severe disease status was predictive of increases in VLA difficulty; few predictors of improvements were identified. CONCLUSION: VLA disability was common, with more disability noted in committed and discretionary activities than in obligatory activities. Because VLA disability has been linked to psychological well-being in previous studies, identification of factors that may protect against such disability is important. PMID- 18383407 TI - Changes in surgical loads and economic burden of hip and knee replacements in the US: 1997-2004. AB - OBJECTIVE: A major component of the economic burden associated with the treatment of arthritis relates to surgical joint replacements of the hips/knees. The purpose of this study was to describe the recent trend of hip/knee replacements and the associated economic burden. METHODS: The Nationwide Inpatient Survey from 1997 to 2004 was analyzed. The International Classification of Diseases, Ninth Revision, Clinical Modification procedure coding was used to identify joint replacement cases. RESULTS: During 2004, approximately 225,900 (95% confidence interval [95% CI] 201,782-250,018) primary hip replacements and 431,485 (95% CI 397,454-465,516) primary knee replacements were performed. This was a 37% increase in primary hip replacements and a 53% increase in primary knee replacements compared with 2000. The number of primary replacement procedures increased equally for males and females; however, the number of procedures increased excessively among persons age 45-64 years. While Medicare remained the major source of payment (55.4% for primary hip replacements, 59.3% for primary knee replacements), private insurance payment experienced a steeper increase. In 2004, the national bill of hospital charges for hip/knee replacements was $26.0 billion, the hospital cost was $9.1 billion, and the amount of reimbursement was $7.2 billion (28% of hospital charges or 79% of hospital cost). Nearly 600,000 hip replacements and 1.4 million knee replacements will be performed in the year 2015. CONCLUSION: The number of joint replacement procedures performed is growing faster than ever. The health care community should consider and prepare for this upcoming demand of surgical loads and its associated economical burden. PMID- 18383408 TI - Validity, reproducibility, and responsiveness of a twelve-joint simplified power doppler ultrasonographic assessment of joint inflammation in rheumatoid arthritis. AB - OBJECTIVE: To investigate the validity, reproducibility, and responsiveness of a simplified power Doppler ultrasound (PDUS) assessment of joint inflammation compared with a comprehensive 44-joint PDUS assessment in patients with rheumatoid arthritis (RA) who started therapy with a biologic agent. METHODS: A total of 160 patients with active RA who started a biologic agent were prospectively recruited in 18 Spanish centers. The patients underwent clinical and laboratory assessment and blinded PDUS examination at baseline and 6 months. A PDUS examination of 128 synovial sites in 44 joints was performed. US synovitis and PD signal were semiquantitatively graded from 1 to 3 in all synovial sites. US count and index for synovitis and PD signal were obtained. PDUS intraobserver and interobserver reliability were evaluated. A process of data reduction based on the frequency of involvement of synovial sites by both synovitis and PD signal was conducted. Construct and discriminant validity of a simplified PDUS assessment was investigated. RESULTS: A PDUS simplified assessment including 24 synovial sites from 12 joints detected 100% of patients with synovitis and 91% of patients with PD signal. There was a highly significant correlation between the 44-joint count and index for synovitis and PD signal and the 12-joint count and index for synovitis and PD signal at baseline and 6 months (r = 0.84-0.90, P < 0.0005). The smallest detectable difference was lower than the mean change in simplified PDUS variables. CONCLUSION: A 12-joint PDUS assessment of RA joint inflammation may be a valid, feasible method for multicenter monitoring of therapeutic response to biologic agents. PMID- 18383409 TI - Long-term followup of temporomandibular joint involvement in juvenile idiopathic arthritis. AB - OBJECTIVE: Temporomandibular joint (TMJ) involvement is a frequent feature in cross-sectional prevalence studies among patients with juvenile idiopathic arthritis (JIA). In this followup study, patients were reviewed after 5 years to study the course of TMJ involvement in relation to disease characteristics. METHODS: Children with JIA from a previous study on TMJ involvement were included. A rheumatologic evaluation including the 6 parameters of the JIA core set and an orthodontic evaluation including an orthopantomogram (OPT) were performed. OPTs were scored according to Rohlin's grading system (grades 0-5). RESULTS: The overall prevalence of patients with condylar alterations decreased from 49% to 40%. Improvement of the alterations was seen in 69% of the initially affected condyles, and consequently improvement was seen in 83% of the initially affected patients. Normalization of the alterations was seen in 67% of the improved condyles, and consequently in 44% of the patients. This proves that the condyle has a regenerative capacity. Improvement was related to low disease activity and a less extensive therapeutic regimen. CONCLUSION: In patients with JIA, condylar alterations can improve and even regenerate. Condylar improvement is associated with a low disease activity. PMID- 18383411 TI - What kind of durometer is best suited for the assessment of skin disease in systemic sclerosis? Comment on the article by Kissin et al. PMID- 18383412 TI - A 32-year-old nursing student with hoarseness and dysphagia to solids. PMID- 18383413 TI - Contemporary prevalence and incidence of work disability associated with rheumatoid arthritis in the US. AB - OBJECTIVE: To provide a contemporary estimate of the prevalence and incidence of rheumatoid arthritis (RA) work disability and examine its permanence over time. METHODS: Data were collected semiannually from 5,384 subjects with rheumatologist diagnosed RA. We examined prevalence in subgroups formed by approximately 5-year disease duration intervals using data from subjects age < or =64 years who were employed at disease onset. Annual incidence was examined longitudinally among subjects who supplied data in 2003, 2004, or 2005, were employed at disease onset and in a year's first survey, and were age < or =63 years. For work disability permanence we used longitudinal data from all subjects who became work disabled and observed whether they later returned to work. RESULTS: Mean age of subjects was 52 years, 82% were female, 63% had more than a high school education, mean disease duration was 14 years, and mean Health Assessment Questionnaire score was 1.0. The prevalence of any premature work cessation was 23% in subjects with 1-3 years duration, 35% in those with 10 years, and 51% in those with > or =25 years RA duration. Arthritis-attributed work cessation was 14%, 29%, and 42%, respectively. Annual incidence of any premature work cessation was approximately 10% and arthritis-attributed work cessation incidence was approximately 6%. Thirty-nine percent of subjects who stopped working later returned to work. CONCLUSION: Work disability prevalence in this sample was high (35% within 10 years disease duration), but may represent a decline from the 50% prevalence reported in 1987. Annual incidence of work disability was higher than prior studies, but the return to work rate was also higher. PMID- 18383414 TI - Impact of ankylosing spondylitis on work and family life: comparisons with the US population. AB - OBJECTIVE: To examine the impact of ankylosing spondylitis (AS) on work disability, nonparticipation in the labor force, marriage, divorce, and childbearing. METHODS: In this cross-sectional survey, we asked AS patients (n = 591, 72.8% men, mean age 48.9 years) from the Los Angeles, Houston, San Francisco, and Washington, DC metropolitan areas about work and family life. The proportion of patients who were work disabled, did not participate in the labor force, had never been married, were divorced, or had a biological child were compared with the proportions expected for each outcome based on data from population surveys. RESULTS: Patients with AS were more likely to be work disabled (13.3% versus 5.7%; P < 0.0001) and somewhat more likely to not participate in the labor force compared with the proportion expected (25.1% versus 21.8%; P = 0.07). These associations were stronger among patients age > or =45 years and those with AS for > or =20 years. AS patients were more likely than expected to have never been married (22.8% versus 15.4%; P < 0.0001) or to be divorced (13.2% versus 10.0%; P = 0.02). Women with AS were less likely than expected to have had children (54.7% versus 64.9%; P = 0.02), but the proportion of men with AS who had children was not different from that of the general population. CONCLUSION: Patients with AS in this study were more likely to have never been married, more likely to be divorced, and more than twice as likely to be work disabled than members of the general population. Women with AS were also less likely to have had children than women in the general population. PMID- 18383415 TI - Comparison of employability outcomes among patients with early or long-standing rheumatoid arthritis. AB - OBJECTIVE: To compare employability between patients with early and long-standing rheumatoid arthritis (RA) and examine the relationships between improvement in employability and disease stage after adjustment for demographic characteristics, disease activity, physical functioning, and response to therapy. METHODS: We evaluated the employability data from 2 double-blind, randomized, placebo controlled studies of infliximab plus methotrexate (MTX) in patients with RA. Patients were incomplete responders to MTX in 1 study and had never taken MTX in the other study. Patients age <65 years were categorized as having early RA (< or =3 years disease duration) or long-standing RA (>3 years disease duration). Physical functioning was assessed using the Health Assessment Questionnaire (HAQ) and clinical response was determined based on the American College of Rheumatology 20% improvement criteria (ACR20). RESULTS: Patients with early RA were more likely to be employable at baseline than those with long-standing RA, even after adjusting for baseline HAQ scores. Among patients who were not employable at baseline but achieved an ACR20 response after 1 year of treatment, after adjusting for baseline HAQ score, the patients with early RA who had never taken MTX were 3 times more likely to become employable compared with those with long-standing RA who had an incomplete response to MTX at baseline. CONCLUSION: In 2 clinical trials, patients with early RA were more likely to show improved employment outcomes after treatment than those with long-standing RA, suggesting intervention as early as possible in the disease course maximizes an individual patient's employment potential. PMID- 18383416 TI - Impaired arterial function associated with thinning of cortical bone in rheumatoid arthritis. AB - OBJECTIVE: To evaluate the association between peripheral arterial function and cortical bone thickness in rheumatoid arthritis (RA). METHODS: In a cross sectional study, we measured the combined cortical thickness (CCT) of the second metacarpal bone from hand radiographs, and the ankle-to-arm systolic blood pressure ratio, also known as ankle-brachial index (ABI), in RA patients. We evaluated the association between the 2 using multinomial logistic regression. RESULTS: We obtained CCT and ABI measurements in 588 RA patients. The mean +/- SD CCT was 3.62 +/- 1.16 mm. The proportion of patients with > or =1 ABI value < or =0.9, indicating obstructed lower limb arteries, increased from 18 (9.2%) of 191 patients in the highest CCT tertile to 25 (12.5%) of 200 in the middle CCT tertile to 38 (19.2%) of 198 in the lowest CCT tertile (P for trend 0.005). We noted a similar pattern for ABI values >1.3, indicative of arterial incompressibility (frequencies in high, middle, and low CCT tertiles were 4.7%, 9.5%, and 19.9%, respectively; P for trend < or =0.001). These trends remained significant after multivariable adjustment for potential confounders. After adjustment for the manifestations of RA and cumulative glucocorticoid dose, the association between CCT and arterial obstruction remained significant, but that with arterial incompressibility weakened considerably. CONCLUSION: There is an association between metacarpal cortical bone thinning and obstruction or incompressibility of the peripheral arteries in RA. The association with incompressibility may be mediated by systemic inflammation and/or glucocorticoids, but that with obstruction is independent of a wide array of potential confounders. Clinicians should be alert to the possibility of impaired arterial function RA patients with thinned metacarpal cortical bone. PMID- 18383417 TI - Fitness, fatigue, disease activity, and quality of life in pediatric lupus. AB - OBJECTIVE: To measure aerobic fitness (maximum oxygen consumption [VO(2peak)]), fatigue, quality of life (QOL), and disease activity in young persons with systemic lupus erythematosus (SLE), and to determine an equation for predicting VO(2peak) from the distance walked in 6 minutes (6MW). METHODS: Fifteen young patients ages 12-19 years with SLE participated. VO(2) was measured by a graded treadmill exercise test. Submaximal exercise intensity was determined from the ventilatory anaerobic threshold. Submaximal aerobic capacity was measured using the 6MW. Patient questionnaires included measures of fatigue, QOL, and physical activity. Physician questionnaires included the Systemic Lupus Erythematosus Disease Activity Index and the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index. RESULTS: Compared with age matched norms, our patients had moderate impairment in aerobic fitness, with a mean +/- SD VO(2peak) of 31.1 +/- 7.9 ml/minute/kg and a mean 6MW distance Z score of -2.4 +/- 2.3. The regression equation to predict VO(2peak) (ml/minute/kg) from 6MW was as follows: 57.1 + [0.038 x distance (meters)] + ( 0.35 x maximal heart rate) (R = 0.67, P = 0.027). Ten subjects (67%) reported significant fatigue. There was no significant correlation of fatigue with fitness measures. Neither fatigue nor fitness was significantly correlated with disease activity, disease damage, or QOL measures. CONCLUSION: Young SLE patients have lower aerobic fitness than reference norms. The 6MW may be used as a marker of fitness, but it is preferable to determine VO(2) with a graded exercise test. Fatigue is a significant symptom in young SLE patients. The application of fatigue measures in young persons is exploratory. The relationship between fatigue and aerobic fitness is not clear. PMID- 18383418 TI - Evaluation of the patient acceptable symptom state as an outcome measure in patients with ankylosing spondylitis: data from a randomized controlled trial. AB - OBJECTIVE: To evaluate the feasibility/acceptability, reliability, external validity, and discriminant capacity of the Patient Acceptable Symptom State (PASS) concept in patients with active ankylosing spondylitis (AS). METHODS: PASS was assessed by asking patients to respond yes or no to a single question: "Considering all the different ways your disease is affecting you, if you would stay in this state for the next months, do you consider that your current state is satisfactory?" during the 24-week, randomized (2:1 ratio), double-blind, placebo-controlled portion of an adalimumab study of 315 patients. RESULTS: PASS reliability was high (kappa = 0.86) in patients with stable disease. Significantly more patients who answered yes to PASS than patients who answered no to PASS were responders based on the ASsessment in Ankylosing Spondylitis (ASAS) International Working Group criteria for 20% improvement (75% who answered yes versus 29% who answered no), the ASAS criteria for 40% improvement (61% versus 16%), the ASAS partial remission criteria (37% versus 3%), and > or =50% improvement on the Bath Ankylosing Spondylitis Disease Activity Index (65% versus 19%; P < 0.001 for all comparisons), demonstrating external validity of the PASS concept. More adalimumab-treated than placebo-treated patients achieved PASS at week 12 (42.3% versus 22.4%; P < 0.001), and more adalimumab-treated than placebo treated patients achieved sustained PASS through week 20 (34.6% versus 12.3%; P < 0.001), indicating excellent discriminant capacity. CONCLUSION: PASS is a feasible, acceptable, reliable, and valid assessment of satisfactory health state in patients with AS. PMID- 18383420 TI - Two aspects of the clinical and humanistic burden of systemic lupus erythematosus: mortality risk and quality of life early in the course of disease. AB - OBJECTIVE: To evaluate mortality risk and predictors among recently diagnosed systemic lupus erythematosus (SLE) patients. METHODS: The vital status of 265 SLE patients and 355 controls enrolled in the Carolina Lupus Study (median time since diagnosis 13 months) was determined approximately 5 years after enrollment. We also assessed the utility of an 8-item quality of life instrument, derived from the standard 36-item Medical Outcomes Study Short Form 36, as an additional measure of disease impact. RESULTS: Five years after diagnosis, 9.7% of patients compared with 0.3% of controls had died (P < 0.0001). Increased mortality risk was seen among older patients (adjusted hazard ratio [HR] 1.03, 95% confidence interval [95% CI] 1.01-1.06 per 1-year increment in age) and among men, African Americans, patients with lupus nephritis, and patients with anti-double-stranded DNA antibodies (adjusted HR approximately 2.0 for each of these factors). In addition, patients who did not provide a blood sample at study enrollment experienced increased mortality risk (age-, sex-, and race-adjusted HR 3.7, 95% CI 1.5-9.1). Similar results were seen in analyses limited to time from study enrollment. Physical component scores of the quality of life measure were 7.7 points lower (P < 0.0001) and mental component scores were 1.8 points lower (P = 0.07) in patients compared with controls. CONCLUSION: The mortality risk among SLE patients is significant, particularly among African Americans, even early in the disease process and even with currently available treatments. Differences between cases and controls in health-related quality of life using the Short Form 8 also demonstrate the multidimensional burden of SLE. PMID- 18383421 TI - Prognostic factors in short-term disability due to musculoskeletal disorders. AB - OBJECTIVE: To identify factors associated with poor outcome in temporary work disability (TWD) due to musculoskeletal disorders (MSDs). METHODS: We conducted a secondary data analysis of a 2-year randomized controlled trial in which all patients with TWD due to MSDs in 3 health districts of Madrid (Spain) were included. Analyses refer to the patients in the intervention group. Primary outcome variables were duration of TWD and recurrence. Diagnoses, sociodemographic, work-related administrative, and occupational factors were analyzed by Cox proportional hazards models. RESULTS: We studied 3,311 patients with 4,424 TWD episodes. The following were independently associated with slower return to work: age (hazard ratio [HR] 0.99, 95% confidence interval [95% CI] 0.98-0.99), female sex (HR 0.84, 95% CI 0.78-0.90), married (HR 0.90, 95% CI 0.83 0.97), peripheral osteoarthritis (HR 0.77, 95% CI 0.6-0.9), sciatica (HR 0.59, 95% CI 0.54-0.65), self-employment (HR 0.56, 95% CI 0.48-0.65), unemployment (HR 0.41, 95% CI 0.28-0.58), manual worker (HR 0.86, 95% CI 0.79-0.94), and work position covered during sick leave (HR 0.84, 95% CI 0.77-0.92). The factors that better predicted recurrence were peripheral osteoarthritis (HR 1.75, 95% CI 1.14 2.6), inflammatory diseases (HR 1.66, 95% CI 1.009-2.72), sciatica (HR 1.30, 95% CI 1.08-1.56), indefinite work contract (HR 1.43, 95% CI 1.14-1.75), frequent kneeling (HR 1.39, 95% CI 1.15-1.69), manual worker (HR 1.19, 95% CI 1.003-1.42), and duration of previous episodes (HR 1.003, 95% CI 1.001-1.005). CONCLUSION: Sociodemographic, work-related administrative factors, diagnosis, and, to a lesser extent, occupational factors may explain the duration and recurrence of TWD related to MSD. PMID- 18383422 TI - Prevalence and clinical correlates of symptoms of depression in patients with systemic sclerosis. AB - OBJECTIVE: To assess the prevalence and predictors of symptoms of depression in a large sample of patients with systemic sclerosis (SSc). METHODS: We conducted a cross-sectional, multicenter study of 376 patients with SSc from the Canadian Scleroderma Research Group Registry. Patients were assessed with the Center for Epidemiologic Studies Depression Scale (CES-D) and through extensive clinical histories and medical examinations. Hierarchical multiple linear regression was used to assess the relationship of sociodemographic and clinical variables with symptoms of depression. RESULTS: The percentages of patients who scored > or =16 and > or =23 on the CES-D were 35.1% and 18.1%, respectively. Patients with less education; patients who were not married; patients with higher physician-rated overall disease severity; and patients with more tender joints, more gastrointestinal symptoms, and more difficulty breathing had significantly higher total CES-D scores. As a group, specific symptom indicators (tender joints, gastrointestinal symptoms, breathing) predicted the most incremental variance in depressive symptoms (DeltaR(2) = 14.2%, P < 0.001) despite being added to the model after demographic, socioeconomic, and global disease duration/severity indicators. CONCLUSION: High levels of depressive symptoms are common in patients with SSc and are related to overall SSc disease severity, as well as specific medical symptoms. Screening for depression among patients with SSc is recommended, although more research is needed to determine the best method for doing this. Successfully treating dyspnea, gastrointestinal symptoms, and joint pain may improve mood, although this has not yet been demonstrated. PMID- 18383423 TI - Determinants of the use of wrist working splints in rheumatoid arthritis. AB - OBJECTIVE: To gain insight into the determinants of the use of wrist working splints among patients with rheumatoid arthritis (RA). METHODS: A qualitative descriptive study was performed among 18 patients with RA who recently received a fabric wrist working splint because of pain due to arthritis of the wrist. Patients were interviewed at home using semistructured in-depth interviews. Interviews were audiotaped and transcribed verbatim and analyzed using the framework approach. RESULTS: The majority of patients indicated that their splint use was dependent on the seriousness of the symptoms (pain, swelling, or tingling feelings) they perceived. Important reasons to wear the splint were reduction of symptoms, wrist support, and immobilization of the wrist. Important reasons to stop wearing the splint were reduced functional abilities using the splint and the performance of dirty or wet activities. CONCLUSION: The reasons for patients to wear and not wear wrist working splints are related to intentional decisions of the patients, which are primarily based on perceived benefits and barriers of splint wearing. The results of this study have been used to develop educational and behavioral strategies to increase adherence to wearing wrist working splints. PMID- 18383424 TI - High titer of serum antiphospholipid antibody levels in adult Henoch-Schonlein purpura and cutaneous leukocytoclastic angiitis. AB - OBJECTIVE: To investigate a possible role of antiphospholipid (aPL) antibodies in adult Henoch-Schonlein purpura (HSP) and cutaneous leukocytoclastic angiitis (CLA). METHODS: We reviewed the records of 30 HSP and 8 CLA adults with an initial cutaneous manifestation of palpable purpura on their lower extremities between 2003 and 2007. Eight microscopic polyangiitis (MPA) patients and 30 healthy persons were recruited as controls. Serum anticardiolipin (aCL), anti phosphatidylserine-prothrombin complex (anti-PS/PT), and anti-beta(2) glycoprotein I (anti-beta(2)GPI) antibody levels in HSP, CLA, MPA patients, and healthy controls were measured by enzyme-linked immunosorbent assay. RESULTS: Twenty-two HSP patients (73%) were positive for serum IgA aCL antibodies. Nineteen (63%) had IgA anti-PS/PT antibodies and 4 (13%) had IgA anti-beta(2)GPI antibodies. IgA aCL and anti-PS/PT antibodies showed a significant correlation (P = 0.007). Twenty (67%) HSP patients had IgM anti-PS/PT antibodies and 6 (20%) had IgG anti-PS/PT antibodies. Six (75%) CLA patients had IgM anti-PS/PT antibodies and 2 (25%) had IgG anti-PS/PT antibodies. In contrast, aPL antibodies were not found in any MPA patients or normal controls. Serum IgA aCL antibody levels in HSP patients showed a significant correlation with serum IgA and C-reactive protein (CRP) levels (P = 0.030 and 0.039, respectively). A positive correlation between CRP and serum IgA anti-PS/PT antibody levels was observed in HSP patients (P = 0.023). Serum IgA aCL antibody levels were also significantly associated with proteinuria according to urinalysis (P = 0.024). CONCLUSION: Serum levels of IgA aCL and anti-PS/PT antibodies were elevated in adult HSP, suggesting that serum IgA antibodies may play some role in adult HSP. IgA aCL and/or anti-PS/PT antibodies could serve as markers for adult HSP and should be monitored as an indicator of adult HSP activity. Small-vessel vasculitis could be dependently associated with the presence of IgM anti-PS/PT antibodies. These findings suggest that aPL antibodies are closely related to the pathogenic factors that trigger the development of vasculitis. PMID- 18383425 TI - Cytomegalovirus UL97 mutations in the era of ganciclovir and maribavir. AB - Mutations in the human CMV UL97 kinase gene are a major mechanism of viral resistance to two anti-CMV drugs, ganciclovir (GCV) and maribavir (MBV). GCV, the most widely used and established therapy for CMV, is a substrate for the UL97 kinase. Well-characterised GCV-resistance mutations at UL97 codons 460, 520 and 590-607 impair the phosphorylation of GCV that is necessary for its antiviral activity, presumably by altering substrate recognition. In contrast, MBV is an inhibitor of the UL97 kinase and is the first new CMV therapy to reach later stage clinical trials in many years. No MBV-resistant CMV isolates have yet been detected in clinical trials, but after culture propagation under drug, UL97 mutations that confer moderate to high-level MBV resistance have been identified at codons 353, 397, 409 and 411. These mutations are located upstream of the GCV resistance mutations and are close to the ATP-binding and catalytic domains common to all kinases, consistent with MBV acting as a small molecule ATP competitive kinase inhibitor. So far, no UL97 mutations are known to confer resistance to both GCV and MBV. PMID- 18383426 TI - Study of the process of secondary failure of sulphonylurea by a Markov model. AB - PURPOSE: The study was to reflect and forecast the evolutive tendency and influence factors of secondary failure of sulphonylurea (SFS) changing with time by using a Markov (MKV) model in the elderly diabetic population in Shanghai. METHODS: A total of 549 patients with elderly diabetes mellitus (DM) were enrolled and grouped in the study. A door-to-door retrospective epidemiological survey was used to collect data. The MKV model was used to assess the process and influence factors of SFS and the MKV process decision support system was adopted to calculate state probability of the MKV process. RESULTS: The rate of SFS in the group of all cases, FPG < or = 10 mmol . L(-1) before treatment and FPG > 10 mmol . L(-1) before treatment, taking single type of sulphonylurea (SU) and taking two types of SU and over respectively was 9.11%, 3.55%, 11.03%, 8.54% and 11.21%. The years of changing into the state of secondary failure in half patients was 5 years, 11-12 years, 4 years, 5 years, 4 years, respectively in the following groups: all cases, FPG < or = 10 mmol . L(-1) before treatment and FPG > 10 mmol . L(-1) before treatment, taking single type of SU and taking two types of SU and over. CONCLUSIONS: A MKV model could predict the long-term evolutive process of SFS by a short-term observation; the speed of SFS was related to the degree of DM patients' condition, patients with higher glucose levels prior to treatment would develop SFS faster; but we cannot postpone the development of secondary failure by exchanging SU types. PMID- 18383427 TI - Host factors for replication and transcription of the influenza virus genome. AB - For replication and transcription of the influenza virus genome of eight segmented and negative-stranded RNAs, not only viral factors but also host derived cellular factors (host factors) are required. This paper focuses on the identification and characterisation of the host factors involved in replication and transcription of the influenza virus genome, reviewing recent progresses in the related molecular mechanisms. Functional assay systems for screening of host factors using cell-free reconstitution systems and an yeast-based influenza virus replicon system are highlighted. We have summarised the property of host factors comprehensively and provided a clue for the perspective in the determination mechanism of host range and virulence and the development of a new strategy to control the influenza virus. PMID- 18383428 TI - The pattern of use of non-steroidal anti-inflammatory drugs (NSAIDs) from 1997 to 2005: a nationwide study on 4.6 million people. AB - PURPOSE: To describe the nationwide pattern of use of non-steroidal anti inflammatory drugs (NSAIDs) in the Danish population. METHODS: All Danish citizens aged 10 or above 1 January 1997 were included in the study. The national prescription registry was used to identify all claimed prescriptions for NSAIDs by the cohort until 2005. By individual-level-linkage of nationwide registries, information was acquired concerning hospitalizations, comorbidity, concomitant pharmacotherapy and socioeconomic factors. RESULTS: The population consisted of 4,614,807 individuals, of which 2,663,706 (57.8%) claimed at least one prescription for NSAID from 1997 to 2005. Ibuprofen and diclofenac were the most frequently used non-selective NSAIDs, whereas rofecoxib and celecoxib were the most frequently used selective cyclooxygenase-2 (COX-2) inhibitors. The usage was similar across all age groups. Female sex and increasing age was associated with increased use of NSAID. Factors predicting extensive NSAID use were: rheumatic disease (odds ratio (OR) = 1.79, 95% confidence interval (CI): 1.69-1.90), gout agents (allopurinol) (OR = 2.54, CI: 2.44-2.64) and other pain medication (OR = 3.27, CI: 3.23-3.31). NSAIDs were most often prescribed for use for one distinct treatment interval and for a short period (overall inter-quartile range [IQR]: 9 66 days). High doses were used in a relatively large proportion of the population (8.9% for etodolac to 19.5% for celecoxib) and 54,373 (2.0%) claimed prescriptions for more than one NSAID at the same time. CONCLUSION: NSAIDs were commonly used in the Danish population. Since NSAIDs have been associated with increased cardiovascular risk, further research on the overall risk associated with these drugs on a national scale is needed. PMID- 18383429 TI - Indicators for preventable drug-related morbidity. Practical application in home based care. AB - AIM: The aim of this study is of evaluating Morris' PDRM indicator system in the Italian Health Care System, especially in the context of home-based health care assistance, so as to assess the reliability of such a system to detect preventable events of drug-related morbidity (DRM). METHODS: This is a pilot study which has been carried out on a control group of patients. The results, which have been obtained, have then been used to calculate the incidence of the preventable drug-related morbidity (PDRM) in the entire population. The sample of patients was taken from the Health District of Chivasso (Local Health Unit 7 in Piedmont). The subjects had all been inserted in the Integrated Home Healthcare Assistance programme in the period from 1st January to 31st December 2004. RESULTS: The PDRM is equal to 32, with an incidence of 15.6% in the entire population (205). Seven indicators out of 19 were able to identify PDRM. Of these, the indicator which showed the best detection rate scored 72%. CONCLUSIONS: The study revealed that this indicator system has the capability of detecting events of PDRM. In addition to this, the version of the system which has been approved by the expert panel has proved executable in the Italian healthcare system, especially in home-based healthcare. PMID- 18383430 TI - Pyrethroids, knockdown resistance and sodium channels. AB - Knockdown resistance to DDT and the pyrethrins was first described in 1951 in the housefly (Musca domestica L.). This trait, which confers reduced neuronal sensitivity to these insecticides, was subsequently shown to confer cross resistance to all synthetic pyrethroid insecticides that have been examined to date. As a consequence, the worldwide commercial development of pyrethroids as a major insecticide class over the past three decades has required constant awareness that pyrethroid overuse has the potential to reselect this powerful resistance mechanism in populations that previously were resistant to DDT. Demonstration of tight genetic linkage between knockdown resistance and the housefly gene encoding voltage-sensitive sodium channels spurred efforts to identify gene mutations associated with knockdown resistance and understand how these mutations confer a reduction in the sensitivity of the pyrethroid target site. This paper summarizes progress in understanding pyrethroid resistance at the molecular level, with particular emphasis on studies in the housefly. PMID- 18383431 TI - Doubly compensated multiplicity-edited HSQC experiments utilizing broadband inversion pulses. AB - We propose a family of doubly compensated multiplicity-edited heteronuclear single quantum coherence (HSQC) pulse sequences. The key difference between our proposed sequences and the compensation of refocusing inefficiency with synchronized inversion sweeps (CRISIS)-HSQC experiments they are based on is that the conventional rectangular 180 degrees pulses on the proton channel in the latter have been replaced by the computer-optimized broadband inversion pulses (BIPs) with superior inversion performance as well as much improved tolerance to B(1) field inhomogeneity. Moreover, all adiabatic carbon 180 degrees pulses during the INEPT and reverse-INEPT periods in the CRISIS-HSQC sequences have also been replaced with the much shorter BIPs, while the adiabatic sweeps during the heteronuclear spin echo for multiplicity editing are kept in place in order to maintain the advantage of the CRISIS feature of the original sequences, namely J independent refocusing of the one-bond (1)H--(13)C coupling constants. These modifications have also been implemented to the preservation of equivalent pathways (PEP)-HSQC experiments. We demonstrate through a detailed comparison that replacing the proton 180 degrees pulses with the BIPs provide additional sensitivity gain that can be mainly attributed to the improved tolerance to B(1) field inhomogeneity of the BIPs. The proposed sequences can be easily adapted for (19)F--(13)C correlations. PMID- 18383432 TI - 13C and 15N NMR chemical shifts of 1-(2',4'-dinitrophenyl) and 1-(2',4',6' trinitrophenyl) pyrazoles in the solid state and in solution. AB - The (13)C and (15)N CPMAS NMR spectra of 18 pyrazoles substituted at position 1 by dinitrophenyl and trinitrophenyl (picryl) groups have been recorded. To help in the assignments, some of these compounds were studied in DMSO-d(6) solution. Phenomena such as the conformation of the N-aryl groups and broadening of splittings due to quadrupolar nuclei are discussed. PMID- 18383433 TI - Bioactive peptides derived from the Limulus anti-lipopolysaccharide factor: structure-activity relationships and formation of mixed peptide/lipid complexes. AB - The design of peptides that would interact and neutralise bacterial endotoxins or LPS could have benefited from the analysis of comparative structure-activity relationships among close-related analogues. Here, we present a comparative structural characterisation of selected peptides derived from the LALF obtained by single-amino-acid replacement, which differ in biological activity. The peptides were characterised in solution using nuclear magnetic resonance, circular dichroism and fluorescence spectroscopies. Membrane mimetic peptide interactions were studied using fluorescence resonance energy transfer with the aid of extrinsic fluorescent probes that allowed the identification of mixed peptide/lipid complexes. PMID- 18383434 TI - NMR study of 5-substituted pyrazolo[3,4-c]pyridine derivatives. AB - Substituted pyrazolopyridines are potent inhibitors of phosphodiesterases and cyclin-dependent kinases. In this study, NMR was used to investigate the potential N1-H and N2-H tautomerism of 5-substituted pyrazolo[3,4-c]pyridine derivatives. Six compounds were fully characterized by using (1)H, (13)C, and (15)N chemical shifts and indirect (1)H--(13)C and (1)H--(15)N coupling constants. The (1)H NMR spectra were measured over a broad range of temperatures. All of the compounds were shown to exist predominantly in the N1-H tautomeric form. Complementary quantum-chemical calculations of the chemical shieldings and indirect spin-spin couplings support the structural conclusions drawn. PMID- 18383435 TI - 119Sn NMR chemical shift tensors in anhydrous and hydrated Si8O20(SnMe3)8 crystals. AB - (119)Sn chemical shift tensors of crystalline trialkyltin functionalized octameric spherosilicates, Si(8)O(20)(SnMe(3))(8), have been determined by fitting sideband intensities in solid-state magic angle spinning (MAS) NMR spectra. Tin chemical shift parameters are exquisitely sensitive to the presence of water of crystallization. Both hydrogen bonding and incipient oxygen-tin bonding from molecular water impact the local tin environment. Tin chemical shift tensors in the crystalline derivatives reflect the changes in geometry and coordination number at the tin centers. Chemical shift correlations on the crystalline derivatives, with known x-ray structures, are used to infer the tin coordination environment in an amorphous sample. PMID- 18383436 TI - T1 relaxation measurement with solvent suppression. AB - Three solvent-suppression pulse sequences of varying complexity were incorporated into the standard inversion recovery pulse program and experimentally evaluated. The least complex suppression sequence involves a composite 90 degrees pulse. A more complex sequence utilizes an excitation sculpting sequence requiring pulsed field gradients, and the most complex sequence incorporates an excitation sculpting sequence with selective rf pulses and gradient pulses. The quality of the spectral data and the accuracy of T(1) measurements of the investigated suppression schemes were evaluated using three aqueous samples with increasing proton content in the water solvent, i.e. by volume 100% D(2)O, 80/20% D(2)O/H(2)O, and 100% H(2)O. For lines removed from the water resonance the T(1) values were generally very consistent between all pulse sequences tested. For lines less than about 200 Hz from the water signal the T(1) measurements become less reliable but are still possible for most of the tested pulse programs. PMID- 18383437 TI - Peptides and pseudopeptides incorporating D-Phe-Pro-Arg and Arg-Gly-Asp lead sequences as potential antithrombotic agents. AB - Peptide leads D-Phe-Pro-Arg for thrombin inhibition and Arg-Gly-Asp for antagonistic activity on fibrinogen receptor were combined in one molecule in order to produce compounds capable of acting both as thrombin inhibitors and as fibrinogen receptor antagonists. Peptide conjugate 7 possessing both leads joined by a tetraglycine linker as well as tripeptides and peptidomimetics with highly overlapped D-Phe-Pro-Arg and Arg-Gly-Asp pharmacophore groups were prepared. Conjugate 7 was found to possess antagonistic activity on fibrinogen receptor, but was unexpectedly inactive as thrombin inhibitor. Compound 9 comprising of highly integrated D-Phe-Pro-Arg and Arg-Gly-Asp pharmacophore groups was found to possess a moderate but well balanced thrombin inhibitory and fibrinogen receptor antagonistic activity. PMID- 18383438 TI - Characterization of nonderivatized plant cell walls using high-resolution solution-state NMR spectroscopy. AB - A recently described plant cell wall dissolution system has been modified to use perdeuterated solvents to allow direct in-NMR-tube dissolution and high resolution solution-state NMR of the whole cell wall without derivatization. Finely ground cell wall material dissolves in a solvent system containing dimethylsulfoxide-d(6) and 1-methylimidazole-d(6) in a ratio of 4:1 (v/v), keeping wood component structures mainly intact in their near-native state. Two dimensional NMR experiments, using gradient-HSQC (heteronuclear single quantum coherence) 1-bond (13)C--(1)H correlation spectroscopy, on nonderivatized cell wall material from a representative gymnosperm pinus taeda (loblolly pine), an angiosperm Populus tremuloides (quaking aspen), and a herbaceous plant Hibiscus cannabinus (kenaf) demonstrate the efficacy of the system. We describe a method to synthesize 1-methylimidazole-d(6) with a high degree of perdeuteration, thus allowing cell wall dissolution and NMR characterization of nonderivatized plant cell wall structures. PMID- 18383439 TI - Peptide fragment of the m3 muscarinic acetylcholine receptor activates G(q) but not G(i2). AB - G(q), a heterotrimeric guanine nucleotide-binding protein, plays important roles such as the regulation of calcium mobilization and cell proliferation. This protein is considered as a promising drug target for the treatment of cardiac hypertrophy. Selective activation of G(q) would be quite useful for analyzing the role of G(q) in signaling pathways. We synthesized m3i3c-a peptide with 16 amino acid residues that corresponds to the junction between the C-terminus of the third intracellular loop and the sixth transmembrane helix (TM-VI) of human m3 muscarinic acetylcholine receptor, which couples to G(q) but not G(i2). At micromolar concentrations, this peptide was found to activate G(q) but not G(i2). This peptide is the first small compound that selectively activates G(q) but not G(i2). PMID- 18383440 TI - Differences in the amount of lipolysis induced by atrial natriuretic peptide in small and large adipocytes. AB - Atrial natriuretic peptide (ANP) is a 28-amino acid polypeptide that is primarily secreted by the heart. ANP is believed to be a hormone that regulates cardiovascular dynamics and renal functions; however, studies conducted in the past few decades revealed that ANP is also a potent lipolytic agent in human adipocytes that functions through the cGMP-dependent pathway. In this study, we separated human adipocytes within the same fat depot into small and large fractions using their floating properties and nylon filters of different pore sizes. Real-time PCR revealed that large adipocytes expressed higher mRNA levels of natriuretic peptide receptor (NPR)-A and hormone sensitive lipase, and binding studies showed that large adipocytes expressed more NPR-A on the membrane than small adipocytes. This finding was confirmed by the increase in the amount of glycerol that was released from adipocytes as the cell size increased. Taken together, these results clearly suggest that adipocyte size is an important determinant of ANP-stimulated lipolysis. PMID- 18383441 TI - Developing tools for the safety specification in risk management plans: lessons learned from a pilot project. AB - PURPOSE: Following the adoption of the ICH E2E guideline, risk management plans (RMP) defining the cumulative safety experience and identifying limitations in safety information are now required for marketing authorisation applications (MAA). A collaborative research project was conducted to gain experience with tools for presenting and evaluating data in the safety specification. This paper presents those tools found to be useful and the lessons learned from their use. METHODS: Archive data from a successful MAA were utilised. Methods were assessed for demonstrating the extent of clinical safety experience, evaluating the sensitivity of the clinical trial data to detect treatment differences and identifying safety signals from adverse event and laboratory data to define the extent of safety knowledge with the drug. RESULTS: The extent of clinical safety experience was demonstrated by plots of patient exposure over time. Adverse event data were presented using dot plots, which display the percentages of patients with the events of interest, the odds ratio, and 95% confidence interval. Power and confidence interval plots were utilised for evaluating the sensitivity of the clinical database to detect treatment differences. Box and whisker plots were used to display laboratory data. CONCLUSIONS: This project enabled us to identify new evidence-based methods for presenting and evaluating clinical safety data. These methods represent an advance in the way safety data from clinical trials can be analysed and presented. This project emphasises the importance of early and comprehensive planning of the safety package, including evaluation of the use of epidemiology data. PMID- 18383442 TI - Risk of ischaemic cardiovascular events from selective cyclooxygenase-2 inhibitors in osteoarthritis. AB - PURPOSE: Most previous observational studies assessing cardiovascular risk associated with Cox-2 inhibitors (Cox-2is) used a case control approach, limiting the assessment of absolute risk by disease group and effect of duration of exposure. We conducted a retrospective cohort study in patients with osteoarthritis. METHODS: Using the Life-link US claims database, all subjects had at least five years history in the database. Exposure was defined as the first chronic period of Cox-2i (celecoxib, rofecoxib or valdecoxib) or naproxen use. Non-users and non-chronic users of non-steroidal anti-inflammatory drug (NSAID)/Cox-2i within the osteoarthritis cohort served as the reference. The primary outcome was myocardial infarction and ischaemic stroke. RESULTS: A cohort of 16,580 subjects were chronically exposed to celecoxib, 9800 received rofecoxib, 2907 received naproxen and 51,539 were non-chronically exposed controls. With a median follow up of 506 days, there were 2116 ischaemic events for the entire cohort. The strongest predictors of AMI/ ischaemic stroke risk were history of ischaemic stroke (HR 2.34, 2.12-2.59) and age 65+ years (HR 2.28, 2.07-2.52). For rofecoxib, (HR 1.25,1.04-1.50), the attributable risk varied from 3 per 1000 patients years in individuals aged under 65 years with no history of CVD to 19 per 1000 patients years in older individuals with a history of CVD. The hazard ratios did not change over time. Celecoxib and naproxen were not associated with increased risks. CONCLUSIONS: The attributable risk for rofecoxib varied substantially with the underlying cardiovascular risk profile, being lower in clinical trial than in clinical practice populations. PMID- 18383443 TI - Coronary heart disease outcomes in patients receiving antidiabetic agents in the PharMetrics database 2000-2007. AB - BACKGROUND: The risk of coronary heart disease (CHD) in users of antidiabetic agents must be quantified to permit reasoned therapeutic choices. OBJECTIVES: To assess the risk of myocardial infarction (MI) and coronary revascularization (CR), in diabetic patients who began rosiglitazone, pioglitazone, metformin, or sulfonylureas. METHODS: We conducted a retrospective cohort study of MI and CR in the PharMetrics database. We performed head-to-head comparisons using propensity score-stratified Cox proportional hazards models, examining risks both on treatment and during total follow-up before regimen switches. RESULTS: For the combined outcome (MI and CR), the crude rates per 1000 person years were 9 on monotherapy, 13 on dual therapy, and 21 on therapies combined with insulin. In the absence of insulin, regimens containing thiazolidinediones (TZDs) tended toward lower risk than comparable regimens containing sulfonylureas and higher risk than those containing metformin. The summary hazard ratio for rosiglitazone versus pioglitazone was 1.04 (95%CI: 0.94-1.14) for total follow-up and 1.05 (0.92-1.19) for on-treatment time. For MI, the hazard ratios were 1.07 (0.89 1.27) for total follow-up and 1.21 (0.95-1.54) for on-treatment time. CONCLUSIONS: The present data indicate that the risk of CHD in patients using TZDs appears to lie between the risks associated with sulfonylureas and metformin. Neither the risk of MI and CR together nor the risk of MI alone was significantly different between rosiglitazone and pioglitazone. A nonsignificant observed excess risk of 21% for MI during on-treatment time will require combination with the results of other studies to provide a reliable assessment. PMID- 18383444 TI - Intracellular trafficking of magnetic nanoparticles to design multifunctional biovesicles. PMID- 18383446 TI - Nearest neighbour adjustment and linear variance models in plant breeding trials. AB - This paper reviews methods for nearest neighbour analysis that adjust for local trend in one dimension. Such methods are commonly used in plant breeding and variety testing. The focus is on simple differencing methods, including first differences and the Papadakis method. We discuss mixed model representations of these methods on the scale of the observed data. Modelling observed data has a number of practical advantages compared to differencing, for example the facility to conveniently compute adjusted cultivar means. Most models considered involve a linear variance-covariance structure and can be represented as state-space models. The reviewed methods and models are exemplified using three datasets. PMID- 18383447 TI - A simple numerical approach towards improving the two one-sided test for average bioequivalence. AB - For testing average bioequivalence, a simple test that improves upon the two-one sided test (TOST) is derived. The new test uses a critical value that is a non increasing continuous function of the sample standard deviation, and the function is numerically obtained. Simulation results show that the resulting test performs well in terms of type I error probability; in particular, the test avoids the conservatism of the TOST. Consequently, the test provides better power compared to the TOST, especially when the variance becomes large. Numerical results also show that our test results in power that is very similar to the other improved tests available in the literature. An advantage of our improved test is that it is just as easy to carry out as the TOST. An example is used to illustrate the new test. PMID- 18383448 TI - Inferences about the between-study variance in meta-analysis with normally distributed outcomes. AB - This paper presents a new approach for confidence interval estimation of the between-study variance in meta-analysis with normally distributed responses based on the concepts of generalized variables. Simulation study shows that the coverage probabilities of the proposed confidence intervals are generally satisfactory. Moreover, the proposed approach can easily provide P -values for hypothesis testing. For meta-analysis of controlled clinical trials or epidemiological studies, within which the responses are normally distributed, the proposed approach is an ideal candidate for making inference about the between study variance. PMID- 18383449 TI - A general approach for sample size and power calculations based on the Haseman Elston method. AB - Risch and Zhang (1995; Science 268: 1584-9) reported a simple sample size and power calculation approach for the Haseman-Elston method and based their computations on the null hypothesis of no genetic effect. We argue that the more reasonable null hypothesis is that of no recombination. For this null hypothesis, we provide a general approach for sample size and power calculations within the Haseman-Elston framework. We demonstrate the validity of our approach in a Monte Carlo simulation study and illustrate the differences using data from published segregation analyses on body weight and heritability estimates on carotid artery artherosclerotic lesions. PMID- 18383452 TI - Neurotoxic implications of the agonistic action of CS-syndrome pyrethroids on the N-type Ca(v)2.2 calcium channel. AB - BACKGROUND: Cismethrin (T-syndrome) and deltamethrin (CS-syndrome) pyrethroids have been previously shown to increase membrane depolarization and calcium influx, but only deltamethrin increased Ca(2+)-dependent neurotransmitter release from rat brain synaptosomes. Deltamethrin's action was blocked by omega-conotoxin GVIA, delineating a separate action at N-type Ca(v)2.2 channels that is consistent with the in vivo release of neurotransmitter. It is hypothesized that other CS-syndrome pyrethroids will elicit similar actions at presynaptic nerve terminals. RESULTS: Nine additional pyrethroids were similarly examined, and these data were used in a cluster analysis. CS-syndrome pyrethroids that possessed alpha-cyano groups, cypermethrin, deltamethrin and esfenvalerate, all caused Ca(2+) influx and neurotransmitter release and clustered with two other alpha-cyano pyrethroids, cyfluthrin and cyhalothrin, that shared these same actions. T-syndrome pyrethroids, bioallethrin, cismethrin and fenpropathrin, did not share these actions and clustered with two non-alpha-cyano pyrethroids, tefluthin and bifenthrin, which likewise did not elicit these actions. Deltamethrin reduced peak current of heterologously expressed wild-type Ca(v)2.2, increased peak current of T422E Ca(v)2.2 and was 20-fold more potent on T422E Ca(v)2.2 than on wild-type channels, indicating that the permanently phosphorylated form of Ca(v)2.2 is the preferred target. CONCLUSIONS: Ca(v)2.2 is directly modified by deltamethrin, but the resulting perturbation is dependent upon its phosphorylation state. The present findings may provide a partial explanation for the different toxic syndromes produced by these structurally distinct pyrethroids. PMID- 18383453 TI - Skeletal tissue engineering using silk biomaterials. AB - Silks have been proposed as potential scaffold materials for tissue engineering, mainly because of their physical properties. They are stable at physiological temperatures, flexible and resist tensile and compressive forces. Bombyx mori (silkworm) cocoon silk has been used as a suture material for over a century, and has proved to be biocompatible once the immunogenic sericin coating is removed. Spider silks have a similar structure to silkworm silk but do not have a sericin coating. This paper provides a general overview on the use of silk protein in biomaterials, with a focus on skeletal tissue engineering. PMID- 18383454 TI - Bone morphogenetic proteins in tissue engineering: the road from laboratory to clinic, part II (BMP delivery). AB - Bone morphogenetic proteins (BMPs) are cytokines with a strong effect on bone and cartilage growth and with important roles during embryonic patterning and early skeletal formation. BMPs have promising potential for clinical bone and cartilage repair, working as powerful bone-inducing components in diverse tissue engineering products. Synthetic polymers, natural origin polymers, inorganic materials and composites may be used as carriers for the delivery of BMPs. Carriers range from nanoparticles to complex three-dimensional (3D) scaffolds, membranes for tissue-guided regeneration, biomimetic surfaces and smart thermosensitive hydrogels. Current clinical uses include spinal fusion, healing of long bone defects and craniofacial and periodontal applications, amongst others. BMP-2 and BMP-7 have recently received approval by the US Food and Drug Administration (FDA) for specific clinical cases, delivered in absorbable collagen sponges. Considering the expanding number of publications in the field of BMPs, there are prospects of a brilliant future in the field of regenerative medicine of bone and cartilage with the use of BMPs. PMID- 18383455 TI - Application of MRI phase-difference mapping to assessment of vascular concentrations of BMS agent in mice. AB - Direct quantitation of contrast agent concentration can be performed using dynamic susceptibility contrast MRI. This method is based on phase imaging and administration of paramagnetic agents such as gadolinium-chelates. This technique has only been applied on humans or primates. However, numerous research models have been developed on small animals like mice. For this reason, the aim of this work was the application of this MRI technique, allowing the direct quantitation of the contrast agent concentrations in vivo, in the mouse vascular system at high field. For this purpose, Dy-DOTA has been preferred to Gd-DOTA due to a lower T(2)* effect. Dy-DOTA shifts in Larmor frequency were measured by phase difference mapping, using fast gradient-echo imaging at short echo times. Such an acquisition sequence allowed the limitation of susceptibility artifacts at high magnetic fields and phase wrapping. As demonstrated in a phantom oriented parallel to the static magnetic field, it is possible to measure contrast agent concentrations between 0 and 10 mm with an uncertainty of about 100 microm. Finally, the method was applied on living mice at 4.7 T. After the bolus injection, the evolution of contrast agent concentrations was assessed in brain blood vessels parallel to B(0). Long-term disappearance of contrast agent was monitored at high spatial resolution every 15 s. Alternatively, lower resolved images at 0.72 s time-resolution allowed preliminary assessment of arterial input functions. The feasibility of quantitative bolus-tracking in small rodents opens the way for comprehensive descriptions of flow and over time-dependent biological processes, especially in pathological murine models. PMID- 18383456 TI - Comments on "Mirror symmetry breaking" of the centrosymmetric CaCO3 crystals with amino acids. PMID- 18383457 TI - A bioluminescent molecular switch for glucose. PMID- 18383458 TI - On-chip synthesis and label-free assays of oligosaccharide arrays. PMID- 18383459 TI - Water-soluble monofunctional perylene and terrylene dyes: powerful labels for single-enzyme tracking. PMID- 18383460 TI - Glycodynamers: fluorescent dynamic analogues of polysaccharides. PMID- 18383461 TI - Selective use of sonographic cervical length measurement for predicting imminent preterm delivery in women with preterm labor and intact membranes. AB - OBJECTIVES: To determine, in a population of women with preterm labor and intact membranes, whether ultrasound cervical length measurement performed only in patients selected according to the Bishop score predicts imminent preterm delivery better than does systematic cervical length measurement in the entire population. METHODS: The Bishop score and sonographic cervical length were recorded prospectively in women with preterm labor between 24 and 34 completed weeks' gestation. Outcome measures were preterm delivery within 48 h and within 7 days. Predictive values were calculated for each marker separately and then in combination. RESULTS: Of the study population of 395 women, 17 (4.3%) and 32 (8.1%) delivered within 48 h and within 7 days, respectively, following inclusion. For delivery within 7 days, areas under the Bishop score (0.848) and sonographic cervical length (0.813) receiver-operating characteristics curves did not differ significantly. For the selective use of sonographic cervical length measurement in patients selected according to the Bishop score, the test was considered positive if the Bishop score was >or= 8, or 4-7 with cervical length 10 lymph nodes was associated with increased overall survival (hazard ratio, 0.52; 95% confidence interval, 0.43-0.64). In addition, with a lymph node density from 0.1% to 12.5% as the referent group, each higher quartile experienced worse survival. CONCLUSIONS: An increased number of lymph nodes removed at the time of cystectomy was associated with improved survival in patients with lymph node-positive bladder cancer. Improved survival was observed at a lower lymph node density threshold than previously reported. The current findings support performing a more extensive lymphadenectomy at the time of cystectomy. PMID- 18383516 TI - The modifying effect of C-reactive protein gene polymorphisms on the association between central obesity and endometrial cancer risk. AB - BACKGROUND: Obesity is a major risk factor for endometrial cancer. Obesity, particularly central obesity, is considered as a systemic inflammatory condition and is related strongly to insulin resistance. C-reactive protein (CRP) is the most recognized biologic marker of chronic systematic inflammation, and it is conceivable that the CRP gene may work together with obesity in the development of endometrial cancer. METHODS: On the basis of a population-based case-control study in a Chinese population, the authors obtained obesity measurements and data on 6 CRP single-nucleotide polymorphisms (SNPs) from 1046 patients with newly diagnosed endometrial cancer (cases) and from 1035 age frequency-matched controls. The association of the CRP SNPs with endometrial cancer risk and their modification on the association between obesity and endometrial cancer risk were evaluated. RESULTS: Although CRP SNPs alone were not associated with endometrial cancer, the associations of endometrial cancer with central obesity, measured as the waist-to-hip ratio (WHR) and the waist circumference, seemed to be stronger in women who were homozygous for the major allele of reference SNP (rs)1130864 (cytidine [C]/C) than in women who had the C/thymidine (T) and T/T genotypes (interaction test: P = .013 for WHR; P = .083 for waist circumference). When the women were stratified further by menopausal status, the observed interactions persisted mainly in premenopausal women (interaction test: P < .001 for WHR; P = .002 for waist circumference). CONCLUSIONS: The current results suggested that, in the Chinese population that was studied, obesity-related insulin resistance and proinflammatory effects may play an important role in endometrial cancer risk, and these effects were modified significantly by the CRP SNP rs1130864. PMID- 18383519 TI - The molecular journey from ductal carcinoma in situ to invasive breast cancer. AB - Ductal carcinoma in situ (DCIS) accounts for approximately 20% of mammographically detected breast cancers. DCIS can recur or progress to invasive breast cancer, but the ability to predict the outcome of patients with DCIS remains limited, leading to inappropriate treatment choices. To the authors' knowledge to date, the hunt for molecular prognostic markers for DCIS has been unsuccessful. Emerging technologies, however, are shedding new light on the biologic course of DCIS. In the current study, the authors review recent findings elucidating the molecular journey from DCIS to invasive cancer and discuss how these findings will lead to more effective treatment with minimization of morbidity. PMID- 18383518 TI - Functional imaging of neuroendocrine tumors with combined PET/CT using 68Ga DOTATATE (DOTA-DPhe1,Tyr3-octreotate) and 18F-FDG. AB - BACKGROUND: The aim was to assess the relevant distribution of the novel PET tracer (68)Ga-DOTATATE in neuroendocrine tumors (NETs) with combined positron emission tomography / computed tomography (PET/CT) and compare its performance with that of (18)F-FDG PET/CT. METHODS: The imaging findings with (68)Ga-DOTATATE and (18)F-FDG on 38 consecutive patients with a diagnosis of primary or recurrent NET were compared and correlated with tumor grade on histology based on ki67 and mitotic index. RESULTS: The sensitivity of (68)Ga-DOTATATE PET/CT was 82% (31 of 38) and that of (18)F-FDG PET/CT was 66% (25 of 38). The sensitivity of combined (68)Ga-DOTATATE and (18)F-FDG PET/CT was 92% (35 of 38). There was greater uptake of (68)Ga-DOTATATE than (18)F-FDG in low-grade NET (median SUV 29 vs 2.9, P < .001). In high-grade NET there was higher uptake of (18)F-FDG over (68)Ga DOTATATE (median SUV 11.7 vs 4.4, P = .03). There was a significant correlation with predominant tumor uptake of (68)Ga-DOTATATE or (18)F-FDG and tumor grade on histology (P < .0001). CONCLUSIONS: (68)Ga-DOTATATE PET/CT is a useful novel imaging modality for NETs and is superior to (18)F-FDG for imaging well differentiated NET. Functional imaging with both (68)Ga-DOTATATE and (18)F-FDG has potential for a more comprehensive tumor assessment in intermediate- and high grade tumors. PMID- 18383520 TI - Secondary osteonecrosis mimicking a femur bone tumor in an adolescent with juvenile idiopathic arthritis. PMID- 18383517 TI - Antiandrogen withdrawal in castrate-refractory prostate cancer: a Southwest Oncology Group trial (SWOG 9426). AB - BACKGROUND: Antiandrogen withdrawal is a potential therapeutic maneuver for patients with progressive prostate cancer. This study was designed to examine antiandrogen withdrawal effects within the context of a large multi-institutional prospective trial. METHODS: Eligibility criteria included progressive prostate adenocarcinoma despite combined androgen blockade. Eligible patients received prior initial treatment with an antiandrogen plus orchiectomy or luteinizing hormone-releasing hormone (LHRH) agonist. Patients were stratified according to type of antiandrogen, type of progression (prostate-specific antigen [PSA] or radiographic), presence or absence of metastatic disease, and prior LHRH agonist versus surgical castration. RESULTS: A total of 210 eligible and evaluable patients had a median follow-up of 5.0 years; 64% of patients previously received flutamide, 32% bicalutamide, and 3% nilutamide. Of the 210 patients, 21% of patients had confirmed PSA decreases of >or=50% (95% CI, 16% to 27%). No radiographic responses were recorded. Median progression-free survival (PFS) was 3 months (95% CI, 2 months to 4 months); however, 19% had 12-month or greater progression-free intervals. Median overall survival (OS) after antiandrogen withdrawal was 22 months (20 and 40 months for those with and without radiographic evidence of metastatic disease, respectively). Multivariate analyses indicated that longer duration of antiandrogen use, lower PSA at baseline, and PSA-only progression at study entry were associated with both longer PFS and OS. Longer antiandrogen use was the only significant predictor of PSA response. CONCLUSIONS: These data indicate a relatively modest rate of PSA response in patients who were undergoing antiandrogen withdrawal; however, PFS can be relatively prolonged (>or=1 year) in approximately 19% of patients. PMID- 18383522 TI - Visilizumab in severe ulcerative colitis: good vision but blurry results. PMID- 18383521 TI - IL23R and IL12B polymorphisms in Spanish IBD patients: no evidence of interaction. AB - BACKGROUND: Recent genomic surveys have identified IL23R and IL12B as susceptibility loci for inflammatory bowel disease (IBD). Our aim in the present study was to ascertain whether the IL23R and IL12B associations with IBD are also observed in our population, and to analyze possible genetic interactions between polymorphisms at IL12B and IL23R, ligand and receptor, respectively. METHODS: In all, 707 IBD patients (344 with Crohn's disease and 363 with ulcerative colitis) and 547 healthy controls from the same ethnic origin (Caucasian Spaniards) were included in the present study. Two single nucleotide polymorphisms (SNPs) in IL23R (rs7517847 and rs11209026) and 2 in IL12B (rs3212227 and rs6887695) genes were analyzed by TaqMan technology. Genetic frequencies were compared with a chi square test. Interaction between genes was analyzed by case-only comparisons. RESULTS: The data show an association of both IL23R SNPs with overall IBD (statistically stronger, rs7517847; odds ratio [OR] = 0.79, 95% confidence interval [CI]: 0.67-0.94, minor allele frequencies: 0.355 in IBD patients versus 0.410 in controls; P = 0.005), somewhat stronger in Crohn's disease (OR = 0.74, 95% CI: 0.61-0.91) than in ulcerative colitis (OR = 0.84, 95% CI: 0.69-1.03). IL12B rs6887695 showed a weak association with IBD (OR = 1.24, 95% CI: 1.04-1.47, minor allele frequencies: 0.375 in IBD patients versus 0.326 in controls, P = 0.012), stronger in UC (OR = 1.31, 95% CI: 1.07-1.60, P = 0.007). No statistically significant differences were apparent when patients were stratified according to clinical characteristics. No interaction was observed between any of the polymorphisms studied at IL12B and IL23R. CONCLUSIONS: Our study confirms the association of IL23R and IL12B with IBD in the Spanish population, but no interaction between either loci could be detected. PMID- 18383523 TI - Has Crohn's disease really occurred anywhere in the digestive tract? PMID- 18383524 TI - Pattern of mucosal tumor necrosis factor-alpha expression in segmental colitis associated with diverticula suggests a truly autonomous clinical entity. PMID- 18383525 TI - Functional reconstruction of complex lip and cheek defect with free composite anterolateral thigh flap and vascularized fascia. AB - BACKGROUND: Extensive composite defects involving the lip and cheek present difficult reconstructive challenges. This study presents a technique using anterolateral thigh (ALT) flaps with vascularized fascia for large complex oral sphincter defect reconstruction. METHODS: Fifteen patients who had undergone oral cancer ablation were enrolled in the study. The average area of intraoral lining and cheek-lip skin defects was 96.9 cm(2). Upper-lip defects ranged 0% to 60%, and lower-lip defects ranged 20% to 80%. Skin and intraoral lining defects were replaced by an ALT fasciacutaneous flap. The vascularized fascia of the flap was used to provide lip suspension. RESULTS: Flap survival was 100%. All but 1 patient had good static suspension. Nine patients had adequate oral competence without drooling, but 6 had occasional oral incontinence. All patients achieved an acceptable appearance. CONCLUSIONS: For extensive cheek-lip composite defects, ALT flap together with vascularized fascia has proven to be a useful option for functional reconstruction. PMID- 18383526 TI - Management of sinonasal undifferentiated carcinoma. AB - BACKGROUND: Our aim was to report the outcomes of treatment for sinonasal undifferentiated carcinoma (SNUC). METHODS: Between September 1992 and October 2005, 15 patients were treated with curative intent with surgery (n=1), surgery and adjuvant radiotherapy (n=9), and definitive radiotherapy (RT) (n=5). Follow up ranged from 11 to 151 months (median, 30); follow-up on living patients ranged from 12 to 151 months (median, 22). No patient was lost to follow-up. RESULTS: Seven patients (47%) developed a recurrence from 3 to 50 months (median, 9) after treatment. The 3-year outcomes were: local control, 78%; locoregional control, 65%; distant metastasis-free survival, 82%; cause-specific survival, 77%, and survival, 67%. The local control rates versus treatment modality were: surgery, 0/1 (0%); surgery and postoperative RT, 7/7 (100%); preoperative RT and surgery, 2/2 (100%); and definitive RT, 2/5 (40%). One patient (7%) treated with surgery and postoperative RT sustained a fatal complication. CONCLUSIONS: Combined surgery and adjuvant RT likely offer the best chance of cure compared with either modality alone. The impact of adjuvant chemotherapy is unclear. PMID- 18383527 TI - Microsomal epoxide hydrolase genotypes and the risk for head and neck cancer. AB - BACKGROUND: Microsomal epoxide hydrolase (mEH) is an enzyme involved in the metabolism of (pre)carcinogens in tobacco smoke. We investigated whether functional genetic polymorphisms in mEH may have a risk-modifying effect on head and neck carcinogenesis. METHODS: Blood from 429 patients with oral, pharyngeal, and laryngeal carcinoma and 419 healthy subjects was investigated for mEH polymorphisms. RESULTS: Logistic regression analysis did not show differences in mEH genotype distributions between patients and controls, when categorized according to predicted mEH enzyme activity. Also no differences were found when evaluated according to tumor localization, sex, or tobacco consumption. A significantly higher incidence of the 139Arg/Arg variant was found in patients with hypopharyngeal carcinoma compared with controls (OR = 4.39, 95% CI = 1.45 to 13.35). CONCLUSION: In contrast to earlier reports, we could not demonstrate a risk-modifying effect of genetic polymorphisms in mEH on head and neck carcinogenesis, except for the predicted high activity variant in patients with hypopharyngeal carcinoma. PMID- 18383528 TI - Osteosarcoma of the head and neck region: lessons learned from a single institution experience of 50 patients. AB - BACKGROUND: Osteosarcoma of the head and neck region is a rare tumor and is a therapeutic challenge because of its aggressive nature and complex anatomical location. Standard management guidelines are lacking due to paucity of published data. METHODS: Fifty patients with head and neck osteosarcoma treated at our institute from 1995 to 2004 were reviewed. RESULTS: There were 32 men and 18 women (median, 30 years). Mandible (56%) was the most common site. Chondroblastic (46%) was the most common histopathological variant. Treatment comprised multimodality approach using surgery (100%), radiotherapy (36%), and chemotherapy (58%). After a median follow-up of 16.6 months, 46% were alive and disease free. Median overall survival was 45.7 months, and progression-free survival was 13.7 months. Mandible and maxilla were favorable sites. Postoperative adjuvant radiotherapy improved local control in patients with adverse prognostic factors. CONCLUSIONS: Surgery remains the mainstay of the treatment of head and neck osteosarcoma. Adjuvant radiotherapy improves outcome in patients with adverse factors. PMID- 18383529 TI - Cystic lymph node metastasis in patients with head and neck cancer: An HPV associated phenomenon. AB - BACKGROUND: : Cystic lymph node metastases have been associated with tonsil cancer. A subset of oropharyngeal cancers contain human papillomavirus (HPV) DNA. The clinical and virologic associations of cystic nodal metastasis in head and neck cancer (HNSCC) were investigated. METHODS: : A retrospective review of patients undergoing neck dissection between 2002 and 2004 identified patients with cystic lymph node metastases. Clinical and pathologic information was recorded. The presence of HPV DNA was analyzed using in situ hybridization in a subset of patients. RESULTS: : One hundred neck dissections were reviewed and 20 had cystic nodes. Seventeen of these patients had primary tumors arising in the palatine or lingual tonsil. Three were "unknown primary." HPV DNA was present in 11 of 13 patients with cystic metastases with tissue available for analysis (87%). No HPV DNA was identified in tumor from 21 patients with solid nodal metastasis (p <.0001). CONCLUSION: : Cystic cervical lymph node metastasis is strongly associated with HPV-related tonsillar HNSCC: (c) 2008 Wiley Periodicals, Inc. Head Neck 2008. PMID- 18383530 TI - Emerging perspectives in epidermal growth factor receptor targeting in head and neck cancer. AB - The epidermal growth factor receptor (EGFR) has been shown to be a promising therapeutic target in head and neck cancer. Cetuximab, a monoclonal antibody against EGFR, has been approved in the United States for use with radiotherapy for head and neck squamous cell carcinoma. However, the role of EGFR targeting agents in other therapeutic modalities, such as combined chemoradiotherapy or induction chemotherapy, remains to be defined. Although results from several clinical trials have demonstrated the therapeutic potentials of EGFR targeting agents in these settings, further studies are necessary before definitive conclusions can be made. The concurrent targeting of EGFR along with other pathways important in carcinogenesis may hold significant therapeutic potential. In particular, several clinical trials are studying the effects of combining agents that target the vascular endothelial growth factor with EGFR inhibitors. Last, studies are ongoing to elucidate the predictive and correlative biomarkers in anti-EGFR therapy to allow for proper patient selection. In the case of cetuximab, these correlative biomarkers may include elements of the immune system in addition to the signal transduction proteins involved in EGFR pathway. PMID- 18383531 TI - The black stuff and Konstantin Nikolaevich Tretiakoff. AB - Konstantin Tretiakoff's doctoral dissertation "Contribution a l'Etude de L'Anatomie pathologique du Locus Niger de Soemmering avec quelques deductions relatives a la pathogenie des troubles du tonus musculaire et De La Maladie de Parkinson" (A Study of the Pathological anatomy of the locus niger of Soemerring and its relevance to the pathogenesis of changes in muscular tone in Parkinson's disease) published in 1919 earned him a silver medal awarded by the University of Paris but failed to gain him the recognition its importance deserved. Despite belated acknowledgment of the importance of his findings Tretiakoff received little acclaim during his life and there have been no biographical accounts written in English or French. Fifty years after his death it seems appropriate to relate some aspects of his interesting peripatetic life and recognize the continuing relevance of his pioneering research on "the black stuff" to our understanding of Parkinson's disease. PMID- 18383532 TI - Antineuronal antibodies in Parkinson's disease. AB - Antineuronal antibodies (ANAs) have been implicated in the pathophysiology of postinfectious movement disorders, such as Sydenham's chorea. However, their relevance in other movement disorders--in the absence of infectious triggers- remains much disputed. We sought to assess the frequency of ANAs in idiopathic Parkinson's disease (IPD) and to explore whether a specific phenotype is associated with the presence of ANAs. For this purpose, we recruited 76 IPD patients, 9 patients with genetic parkinsonism, and 10 with one of the parkinson plus syndromes. They were all subjected to a comprehensive clinical review. In addition, 50 patients with non-extrapyramidal neurological disease and 30 healthy blood donors served as control populations. Blood samples were tested for the presence of ANAs with Western blotting, using recombinant proteins of the three putative antigens (aldolase C, neuron-specific enolase, and pyruvate kinase M1). We found these antibodies in 11.8% of the 76 IPD patients, which differed significantly from healthy controls (0%, P = 0.043), but nonsignificantly from patients with genetic parkinsonism (11.1%), with a parkinson-plus syndrome (10%), or from neurological disease controls (4%). With respect to relevant disease characteristics, IPD patients with or without ANAs were indistinguishable, except for atypical disease features (mainly early falls or freezing and marked Pisa syndrome), which were more frequent in the ANA-positive IPD group. We conclude that ANAs do not play a role in the majority of patients with IPD, but might be relevant in the pathogenesis of IPD with atypical features. PMID- 18383533 TI - Oculomotor function in multiple system atrophy: clinical and laboratory features in 30 patients. AB - We reviewed the clinical and laboratory oculomotor features in 30 patients with probable multiple system atrophy (MSA), 22 with MSA-P and 8 with MSA-C. Six patients were also examined post mortem, MSA being confirmed in four and excluded in two (Parkinson's disease and progressive supranuclear palsy). Clinical examination showed the following abnormalities; excessive square wave jerks--21 of 30 patients; mild vertical supranuclear gaze palsy--8 of 30; gaze-evoked nystagmus--12 of 30 patients, three of whom had no extraocular evidence of cerebellar dysfunction; positioning downbeat nystagmus--10 of 25; mild or moderate saccadic hypometria--22 of 30; impaired ("broken up") smooth pursuit--28 of 30; reduced VOR suppression--16 of 24. Electro-oculography and caloric testing did not add significant extra information. In patients presenting with an akinetic-rigid syndrome it can be difficult to differentiate idiopathic Parkinson's disease from MSA-P and other causes of atypical parkinsonism. Our findings suggest that the presence of excessive square wave jerks, mild-moderate hypometria of saccades, impaired VOR suppression, spontaneous nystagmus or positioning downbeat nystagmus may be oculomotor "red flags" or clues to the presence of MSA. Further, the presence of clinically slow saccades, or moderate to-severe gaze restriction, suggests a diagnosis other than MSA. PMID- 18383534 TI - Neuronal activity of the red nucleus in Parkinson's disease. AB - Precise placement of the electrodes for stimulation of the subthalamic nucleus (STN) in Parkinson's disease (PD) is crucial for the therapeutic benefit. As a result of the mistargeting and misplacement of the electrodes during surgery in 2 patients with PD, we have characterized the neuronal firing in the red nucleus (RN) and observed the effects of stimulation of this nucleus. Although the neuronal firing (mean +/- SD) of the RN (34 +/- 4.4 Hz) resembles that described for the STN (33.1 +/- 16.6 Hz), a higher proportion of cells responded to the movement of the contralateral limbs (70-80%). Stimulation in the area of the RN induced intolerable side effects without motor improvement. We conclude that the STN and RN have some similar neurophysiological features but can be distinguished intraoperatively. This initial description of the physiological characteristics of the RN in humans will draw attention to the possibility of confusing the RN and STN during intraoperative recording. PMID- 18383535 TI - Atypical Parkinsonism in distal myopathy with rimmed vacuoles. AB - A patient with distal myopathy with rimmed vacuoles (DMRV) exhibited Parkinsonism with a severe writing tremor that responded poorly to levodopa. Molecular genetic analysis revealed that the patient had the D176V/V572L compound heterozygous mutation in the UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) gene. Histopathological examination of a biopsied muscle specimen yielded findings compatible with those of DMRV, which is characterized by the presence of rimmed vacuoles without inflammatory cell infiltration in muscle fibers. The finding of normal cardiac meta-iodobenzylguanide uptake makes the possibility of incidental Parkinson's disease in this patient unlikely. These observations raise the possibility that atypical Parkinsonism is a rare complication of DMRV associated with GNE mutation. PMID- 18383536 TI - Evidence of increased odds of essential tremor in Parkinson's disease. AB - In a case control study using a standardized protocol, 600 subjects were evaluated for essential tremor (ET). We demonstrated that ET was significantly more frequent in patients with Parkinson's disease (PD) (12/204, 5.9%) compared to diseased controls (2/206, 1%) and healthy controls (1/190, 0.5%). A regression analysis with ET as outcome and group (either PD or healthy controls or diseased controls) as independent variable (adjusting for age and sex) revealed that PD had higher odds of having ET than diseased controls (OR = 5.43, 95% CI = 1.16, 25.39, P < 0.001) and healthy controls (OR = 10.87, 95% CI = 1.39, 85.15, P < 0.001). The low frequency of ET in our controls was further confirmed in a follow up study in a group of age and gender matched general medical patients who attended an outpatient clinic (0% frequency). Eight of 204 PD (3.9%) compared to none of diseased (0%) (P = 0.004) and healthy controls (0%) (P = 0.008) had a prior diagnosis of ET. The duration of ET symptoms in patients with PD was 25.1 +/- 19.6 (range 3-60) years. A multivariate analysis demonstrated that a lower dose of levodopa (OR = 0.993, 95%CI for OR = 0.988, 0.997, P < 0.001) and a higher age of onset of disease (OR = 1.108, 95%CI for OR = 1.035, 1.187, P < 0.001) were associated with increased odds of PD with ET, compared to patients with PD without ET. In our Asian population, patients with PD were 5 to 10 times more likely to have ET compared to diseased and healthy controls, suggesting that the association of ET and PD is unlikely to be ethnicity-specific. PMID- 18383537 TI - Provocation of Parkinsonian tremor. AB - Classical Parkinsonian rest tremor typically fluctuates over time and can be provoked by stressful situations. We quantified and compared the influence of different provocation methods on classical rest tremor severity. The effect of counting backwards from 100, tapping of the contralateral foot and a Stroop test on the Unified Parkinson's Disease Rating Scale (UPDRS) III rest tremor scores and the accelerometrically measured tremor amplitudes (total power) were analyzed in 18 patients with Parkinson's disease and a Type I tremor. Each of the three provocation methods increased the UPDRS III rest tremor score by 1-2 and the total power by 1-2 orders of magnitude compared with baseline (P < 0.001). The maximal effect was reached on average after 2-3 minutes of provocation. The effects were not significantly different. Provocations clearly influence the result of clinical rest tremor ratings, with the kind of provocation being of minor importance. We therefore suggest that each assessment of Parkinsonian rest tremor should include a systematic provocation and this should be formally included in future versions of the UPDRS. PMID- 18383539 TI - Golimumab in patients with active rheumatoid arthritis despite treatment with methotrexate: a randomized, double-blind, placebo-controlled, dose-ranging study. AB - OBJECTIVE: To assess the efficacy, safety, and pharmacology of subcutaneous administration of golimumab in patients with active rheumatoid arthritis (RA) despite treatment with methotrexate (MTX). METHODS: Patients were randomly assigned in a double-blinded manner to receive injections of placebo plus MTX or 50 mg or 100 mg golimumab every 2 or 4 weeks plus MTX through week 48. Patients originally assigned to receive injections every 2 weeks had the interval increased to every 4 weeks starting at week 20. The primary end point was the proportion of patients meeting the American College of Rheumatology 20% improvement criteria (achieving an ACR20 response) at week 16. The study was powered to detect a difference in the primary end point when the combined golimumab groups and at least 1 of the individual dose groups were compared with placebo. RESULTS: The primary end point was attained. Sixty-one percent of patients in the combined golimumab plus MTX dose groups achieved an ACR20 response at week 16 compared with 37% of patients in the placebo plus MTX group (P=0.010). In addition, 79% of patients in the group receiving 100 mg golimumab every 2 weeks achieved an ACR20 response (P<0.001 versus placebo). Through week 20 (after which patients receiving placebo were switched to active infliximab therapy), serious adverse events were reported in 9% of patients in the combined golimumab groups and in 6% of patients in the placebo group. CONCLUSION: Golimumab plus MTX effectively reduces the signs and symptoms of RA and is generally well tolerated in patients with an inadequate response to MTX. PMID- 18383541 TI - MR imaging methods for assessing fetal brain development. AB - Fetal magnetic resonance imaging provides an ideal tool for investigating growth and development of the brain in vivo. Current imaging methods have been hampered by fetal motion but recent advances in image acquisition can produce high signal to noise, high resolution 3-dimensional datasets suitable for objective quantification by state of the art post acquisition computer programs. Continuing development of imaging techniques will allow a unique insight into the developing brain, more specifically process of cell migration, axonal pathway formation, and cortical maturation. Accurate quantification of these developmental processes in the normal fetus will allow us to identify subtle deviations from normal during the second and third trimester of pregnancy either in the compromised fetus or in infants born prematurely. PMID- 18383542 TI - Introduction: dynamic imaging of the developing nervous system. PMID- 18383540 TI - In vivo imaging of presynaptic terminals and postsynaptic sites in the mouse submandibular ganglion. AB - Much of what is currently known about the behavior of synapses in vivo has been learned at the mammalian neuromuscular junction, because it is large and accessible and also its postsynaptic acetylcholine receptors (AChRs) are readily labeled with a specific, high-affinity probe, alpha-bungarotoxin (BTX). Neuron neuron synapses have thus far been much less accessible. We therefore developed techniques for imaging interneuronal synapses in an accessible ganglion in the peripheral nervous system. In the submandibular ganglion, individual preganglionic axons establish large numbers of axo-somatic synapses with postganglionic neurons. To visualize these sites of synaptic contact, presynaptic axons were imaged by using transgenic mice that express fluorescent protein in preganglionic neurons. The postsynaptic sites were visualized by labeling the acetylcholine receptor (AChR) alpha7 subunit with fluorescently tagged BTX. We developed in vivo methods to acquire three-dimensional image stacks of the axons and postsynaptic sites and then follow them over time. The submandibular ganglion is an ideal site to study the formation, elimination, and maintenance of synaptic connections between neurons in vivo. PMID- 18383543 TI - Imaging the spatiotemporal organization of neural activity in the developing spinal cord. AB - In this review, we discuss the use of imaging to visualize the spatiotemporal organization of network activity in the developing spinal cord of the chick embryo and the neonatal mouse. We describe several different methods for loading ion- and voltage-sensitive dyes into spinal neurons and consider the advantages and limitations of each one. We review work in the chick embryo, suggesting that motoneurons play a critical role in the initiation of each cycle of spontaneous network activity and describe how imaging has been used to identify a class of spinal interneuron that appears to be the avian homolog of mammalian Renshaw cells or 1a-inhibitory interneurons. Imaging of locomotor-like activity in the neonatal mouse revealed a wave-like activation of motoneurons during each cycle of discharge. We discuss the significance of this finding and its implications for understanding how locomotor-like activity is coordinated across different segments of the cord. In the last part of the review, we discuss some of the exciting new prospects for the future. PMID- 18383545 TI - Optical imaging of infants' neurocognitive development: recent advances and perspectives. AB - Near-infrared spectroscopy (NIRS) provides a unique method of monitoring infant brain function by measuring the changes in the concentrations of oxygenated and deoxygenated hemoglobin. During the past 10 years, NIRS measurement of the developing brain has rapidly expanded. In this article, a brief discussion of the general principles of NIRS, including its technical advantages and limitations, is followed by a detailed review of the role played so far by NIRS in the study of infant perception and cognition, including language, and visual and auditory functions. Results have highlighted, in particular, the developmental changes of cerebral asymmetry associated with speech acquisition. Finally, suggestions for future studies of neurocognitive development using NIRS are presented. Although NIRS studies of the infant brain have yet to fulfill their potential, a review of the work done so far indicates that NIRS is likely to provide many unique insights in the field of developmental neuroscience. PMID- 18383544 TI - The adolescent brain: insights from functional neuroimaging research. AB - With the development of functional neuroimaging tools, the past two decades have witnessed an explosion of work examining functional brain maps, mostly in the adult brain. Against this backdrop of work in adults, developmental research begins to gather a substantial body of knowledge about brain maturation. The purpose of this review is to present some of these findings from the perspective of functional neuroimaging. First, a brief survey of available neuroimaging techniques (i.e., fMRI, MRS, MEG, PET, SPECT, and infrared techniques) is provided. Next, the key cognitive, emotional, and social changes taking place during adolescence are outlined. The third section gives examples of how these behavioral changes can be understood from a neuroscience perspective. The conclusion places this functional neuroimaging research in relation to clinical and molecular work, and shows how answers will ultimately come from the combined efforts of these disciplines. PMID- 18383547 TI - Rapid activity-dependent delivery of the neurotrophic protein CPG15 to the axon surface of neurons in intact Xenopus tadpoles. AB - CPG15 (aka neuritin) is an activity-induced GPI-anchored axonal protein that promotes dendritic and axonal growth, and accelerates synaptic maturation in vivo. Here we show that CPG15 is distributed inside axons and on the axon surface. CPG15 is trafficked to and from the axonal surface by membrane depolarization. To assess CPG15 trafficking in vivo, we expressed an ecliptic pHluorin (EP)-CPG15 fusion protein in optic tectal explants and in retinal ganglion cells of intact Xenopus tadpoles. Depolarization by KCl increased EP CPG15 fluorescence on axons. Intraocular kainic acid (KA) injection rapidly increased cell-surface EP-CPG15 in retinotectal axons, but coinjection of TTX and KA did not. Consistent with this, we find that intracellular CPG15 is localized to vesicles and endosomes in presynaptic terminals and colocalizes with synaptic vesicle proteins. The results indicate that the delivery of the neurotrophic protein CPG15 to the axon surface can be regulated on a rapid time scale by activity-dependent mechanisms in vivo. PMID- 18383548 TI - Two-photon imaging of dendritic spine development in the mouse cortex. AB - Dendritic spines are the postsynaptic sites of most excitatory synapses in the mammalian brain. With the advent of two-photon microscopy and transgenic mice expressing fluorescent proteins, dendritic spines can now be imaged in the living cerebral cortex over time scales ranging from seconds to years. Recent studies with this in vivo imaging approach have begun to provide important insights into the development and plasticity of dendritic spines in the intact brain. Here, we review these studies and discuss technical requirements for image acquisition. We envision that intravital two-photon imaging at the level of individual synapses will greatly expand our current understandings of how neuronal networks are assembled and modified throughout life. PMID- 18383546 TI - Using imaging and genetics in zebrafish to study developing spinal circuits in vivo. AB - Imaging and molecular approaches are perfectly suited to young, transparent zebrafish (Danio rerio), where they have allowed novel functional studies of neural circuits and their links to behavior. Here, we review cutting-edge optical and genetic techniques used to dissect neural circuits in vivo and discuss their application to future studies of developing spinal circuits using living zebrafish. We anticipate that these experiments will reveal general principles governing the assembly of neural circuits that control movements. PMID- 18383549 TI - Molecular MALDI imaging: an emerging technology for neuroscience studies. AB - Mass spectrometry (MS) has become an essential tool for the detection, identification, and characterization of the molecular components of biological processes, such as those responsible for the dynamic properties of the nervous system. Generally, the application of these powerful techniques requires the destruction of the specimen under study, but recent technological advances have made it possible to apply the matrix-assisted laser desorption/ionization (MALDI) MS technique directly to tissue sections. The major advantage of direct MALDI analysis is that it enables the acquisition of local molecular expression profiles, while maintaining the topographic integrity of the tissue and avoiding time-consuming extraction, purification, and separation steps, which have the potential for introducing artifacts. With automation and the ability to display complex spectral data using imaging software, it is now possible to create multiple 2D maps of selected biomolecules in register with tissue sections, a method now known as MALDI Imaging, or MSI (for Mass Spectrometry Imaging). This creates, for example, an opportunity to correlate functional states, determined a priori with live recording or imaging, with the corresponding molecular maps obtained at the time the tissue is frozen and analyzed with MSI. We review the increasing application of MALDI Imaging to the analysis of molecular distributions of proteins and peptides in nervous tissues of both vertebrates and invertebrates, focusing in particular on recent studies of neurodegenerative diseases and early efforts to implement assays of neuronal development. PMID- 18383550 TI - From synapses to behavior: development of a sensory-motor circuit in the leech. AB - The development of neuronal circuits has been advanced greatly by the use of imaging techniques that reveal the activity of neurons during the period when they are constructing synapses and forming circuits. This review focuses on experiments performed in leech embryos to characterize the development of a neuronal circuit that produces a simple segmental behavior called "local bending." The experiments combined electrophysiology, anatomy, and FRET-based voltage-sensitive dyes (VSDs). The VSDs offered two major advantages in these experiments: they allowed us to record simultaneously the activity of many neurons, and unlike other imaging techniques, they revealed inhibition as well as excitation. The results indicated that connections within the circuit are formed in a predictable sequence: initially neurons in the circuit are connected by electrical synapses, forming a network that itself generates an embryonic behavior and prefigures the adult circuit; later chemical synapses, including inhibitory connections, appear, "sculpting" the circuit to generate a different, mature behavior. In this developmental process, some of the electrical connections are completely replaced by chemical synapses, others are maintained into adulthood, and still others persist and share their targets with chemical synaptic connections. PMID- 18383552 TI - Using voltage-sensitive dye recording to image the functional development of neuronal circuits in vertebrate embryos. AB - Recent developments in the design of voltage-sensitive dyes and of recording apparatuses for detecting voltage-dependent changes in the optical properties of such dyes have established voltage-sensitive dye recording as an important technique for assessing the functional development of neuronal circuits in the brain and spinal cord. Here we discuss general technical issues regarding the recording of voltage-sensitive dye signals and describe studies that have utilized this approach to follow the development of sensory and sensorimotor circuits in the embryonic brain stem. Functional imaging through voltage sensitive dye recording permits a noninvasive analysis of synaptic development and function at submillisecond temporal resolution in widely distributed circuits. These advantages are particularly valuable in assessing sensorimotor circuit development at early stages when neurons are small and synapses are fragile. PMID- 18383551 TI - Imaging second messenger dynamics in developing neural circuits. AB - A characteristic feature of developing neural circuits is that they are spontaneously active. There are several examples, including the retina, spinal cord, and hippocampus, where spontaneous activity is highly correlated among neighboring cells, with large depolarizing events occurring with a periodicity on the order of minutes. One likely mechanism by which neurons can "decode" these slow oscillations is through activation of second messenger cascades that either influence transcriptional activity or drive posttranslational modifications. Here, we describe recent experiments where imaging has been used to characterize slow oscillations in the cAMP/PKA second messenger cascade in retinal neurons. We review the latest techniques in imaging this specific second messenger cascade, its intimate relationship with changes in intracellular calcium concentration, and several hypotheses regarding its role in neurodevelopment. PMID- 18383553 TI - Inhibition of HBV gene expression and replication by stably expressed interferon alpha1 via adeno-associated viral vectors. AB - BACKGROUND: Interferon-alpha2 (IFNalpha2) is routinely used for anti-hepatitis B virus (HBV) treatment. However, the therapeutic efficiency is unsatisfactory, particularly in East Asia. Such inefficiency might be a result of the short half life, relatively low local concentration and strong side-effects of interferons. Frequent and repeated injection is also a big burden for patients. In the present study, a single dose of vector-delivered IFNalpha1 was tested for its anti-HBV effects. METHODS: Adeno-associated viral vector (AAV-IFNalpha1) was generated to deliver the IFNalpha1 gene into hepatocytes. IFNalpha1, hepatitis B surface (HBsAg) and e (HBeAg) antigens were measured by enzyme-linked immunosorbent assay and/or western blotting. The level of viral DNA was measured by quantitative real time polymerase chain reaction. RESULTS: AAV-IFNalpha1 effectively transduced HBV producing cells (HepAD38) and mouse hepatocytes, where IFNalpha1 was expressed in a stable manner. Both intracellular and extracellular HBsAg and HBeAg were significantly reduced in vitro. In the HBV-producing mice, the concentration of IFNalpha1 in the liver was eight-fold higher than that in plasma. Compared with control groups, HBeAg/HBsAg antigen levels were reduced by more than ten-fold from day 1-5, and dropped to an undetectable level on day 9 in the AAV-IFNalpha1 group. Concurrently, the level of viral DNA decreased over 30-fold for several weeks. CONCLUSIONS: A single dose administration of AAV-IFNalpha1 viral vector displayed prolonged transgene expression and superior antiviral effects both in vitro and in vivo. Therefore, the use of AAV-IFNalpha1 might be a potential alternative strategy for anti-HBV therapy. PMID- 18383554 TI - Human mastoid periosteum-derived stem cells: promising candidates for skeletal tissue engineering. AB - Currently, mesenchymal stem cells (MSCs) are considered as the most eligible cells for skeletal tissue engineering. However, factors such as difficult stimulation and control of differentiation in vivo hamper their clinical use. In contrast, periosteum or periosteum-derived cells (PCs) are routinely clinically applied for bone and cartilage repair. PCs have often been named MSCs but, although cells of osteochondrogenic lineages arise from MSCs, it is unclear whether periosteum really contains MSCs. Our aim was to investigate the MSC-like character of PCs derived from the periosteum of mastoid bone. Harvesting of periosteum from mastoid bone is easy, so mastoid represents a good source for the isolation of PCs. Therefore, we analysed the MSC-like growth behaviour and the expression of embryonic, ectodermal, endodermal and mesodermal markers by microarray and FACS technology, and the multilineage developmental capacity of human PCs. Regarding clinical relevance, experiments were performed in human serum-supplemented medium. We show that PCs do not express early embryonic stem cell markers such as Oct4 and Nanog, or the marker of haematopoietic stem cells CD34, but express some other MSC markers. Osteogenesis resulted in the formation of calcified matrix, increased alkaline phosphatase activity, and induction of the osteogenic marker gene osteocalcin. Staining of proteoglycans and deposition of type II collagen documented chondrogenic development. As shown for the first time, adipogenic stimulation of mastoid-derived PCs resulted in the formation of lipid droplets and expression of the adipogenic marker genes aP2 and APM1. These results suggest MSC-like PCs from mastoid as candidates for therapy of complex skeletal defects. PMID- 18383555 TI - Monitoring of FLT3 phosphorylation status and its response to drugs by flow cytometry in AML blast cells. AB - FLT3 mutation and overexpression in most acute myeloid leukaemia (AML) patients make this tyrosine kinase receptor an attractive therapeutic target. FLT3 kinase inhibitors are actually in clinical trials, thus it is critical to develop a reproducible and standardized method for screening of FLT3 activation and for monitoring its inhibition in response to drug in AML patients. We developed a flow cytometry method to analyse phosphorylated FLT3 (P-FLT3) in samples with <10(5) cells. The method was first validated in FLT3 wild-type (HL60/WT) and mutant (MV4-11/ITD(+)) as well as FLT3 negative (K562) cell lines. The method also proved to be reproducible in AML patient samples. Analysis was performed after exposure to drugs (CEP-701 and SU11657), in vitro and in vivo. In response to increasing drug concentrations, there was a linear reduction in P-FLT3. Intracellular flow cytometry analysis correlated with Western blot and XTT assays; flow cytometry data also correlated with FLT3 mutational status. The results highlight a rapid method to detect P-FLT3 protein at the single cell level by flow cytometry which enables an accurate assessment of FLT3 kinase activity in blast cells in response to novel tyrosine kinase inhibitors. PMID- 18383556 TI - Computer-aided diagnosis using morphological features for classifying breast lesions on ultrasound. AB - OBJECTIVES: To develop and evaluate a computer-aided diagnosis (CAD) system with automatic contouring and morphological analysis to aid in the classification of breast tumors using ultrasound. METHODS: We evaluated 118 breast lesions (34 malignant and 84 benign tumors). Each tumor contour was automatically extracted from the digitized ultrasound image. Nineteen practical morphological features from the extracted contour were calculated and principal component analysis (PCA) was applied to find independent features. A support vector machine (SVM) classifier utilized the selected principal vectors to identify the breast tumor as benign or malignant. In this study, all the cases were sampled with k-fold cross-validation (k = 10) to evaluate the performance by receiver-operating characteristics (ROC) curve analysis. RESULTS: The areas under the ROC curves for the proposed CAD systems using all morphological features and the lower dimensional principal vector were 0.91 and 0.90, respectively. The classification ability for breast tumors using morphological information was good. CONCLUSIONS: This system differentiates benign from malignant breast tumors well and therefore provides a clinically useful second opinion. Moreover, the morphological features are nearly setting-independent and thus available to various ultrasound machines. PMID- 18383557 TI - Conditioned media cause increases in select osteogenic and adipogenic differentiation markers in mesenchymal stem cell cultures. AB - The optimal mesenchymal stem cell (MSC) culture conditions that would allow clinically viable tissue-engineered devices are still yet to be determined. Most MSCs are found in the bone marrow, an area that also contains numerous osteoblasts and adipocytes. Paracrine signalling may be leveraged to modulate MSC differentiation in the preparation of a tissue-engineered device. Thus, the objectives of this study were to determine the effects of adipocyte-conditioned medium (CM) on MSC differentiation to osteoblasts and to determine the effects of osteoblast CM on MSC differentiation to adipocytes. Two groups of murine MSCs were given either an osteogenic differentiation medium or an adipogenic differentiation medium. CM was taken from one group and administered to the opposite group in concentrations of 25% or 50%. Metabolic activity, total protein and alkaline phosphatase (ALP) assays were conducted on the osteogenic group at predefined time points throughout the 21 day study, while metabolic activity, triglyceride and oil red O assays were conducted on the adipogenic group at predefined time points. Adipocyte CM administered at a concentration of 50% increased the ALP production of MSCs undergoing osteogenic differentiation. Additionally, osteoblast CM increased the triglyceride production of MSCs undergoing adipogenic differentiation and enlarged the lipid vesicles that were produced by the cells. The effects of the osteoblast CM were seen at both concentrations, but were greatest at the 50% CM level. PMID- 18383558 TI - 68Ga-PET: a powerful generator-based alternative to cyclotron-based PET radiopharmaceuticals. AB - PET (positron emission tomography) is a powerful diagnostic and imaging technique which requires short-lived positron emitting isotopes. The most commonly used are accelerator-produced (11)C and (18)F. An alternative is the use of metallic positron emitters. Among them (68)Ga deserves special attention because of its availability from long-lived (68)Ge/(68)Ga generator systems which render (68)Ga radiopharmacy independent of an onsite cyclotron. The coordination chemistry of Ga(3+) is dominated by its hard acid character. A variety of mono- and bifunctional chelators have been developed which allow the formation of stable (68)Ga(3+)complexes and convenient coupling to biomolecules. (68)Ga coupling to small biomolecules is potentially an alternative to (18)F- and (11)C-based radiopharmacy. In particular, peptides targeting G-protein coupled receptors overexpressed on human tumour cells have shown preclinically and clinically high and specific tumour uptake. Kit-formulated precursors along with the generator may be provided, similar to the (99)Mo/(99m)Tc-based radiopharmacy, still the mainstay of nuclear medicine. PMID- 18383559 TI - Type 1 diabetes: can exercise impair the autoimmune event? The L arginine/glutamine coupling hypothesis. AB - Prevention of type 1 diabetes mellitus (T1DM) requires early intervention in the autoimmune process directed against beta-cells of the pancreatic islets of Langerhans, which is believed to result from a disorder of immunoregulation. According to this concept, a T-helper lymphocyte of type 1 (Th1) subset of T lymphocytes and their cytokine products, the type 1 cytokines [e.g. interleukin 2 (IL-2), interferon gamma (IFN-gamma) and tumour necrosis factor beta (TNF-beta)] prevail over immunoregulatory (anti-inflammatory) Th2 subset and its cytokine products, i.e. type 2 cytokines (e.g. IL-4, IL-6 and IL-10). This allows type 1 cytokines to initiate a cascade of immune/inflammatory processes in the islet (insulitis), culminating in beta-cell destruction. Activation of sympathetic corticotropin-releasing hormone (CRH) axis by psychological stress induces specifically Th1 cell overactivity that determines enhanced glutamine utilization and consequent poor L-arginine supply for nitric oxide (NO)-assisted insulin secretion. This determines the shift of intraislet glutamate metabolism from the synthesis of glutathione (GSH) to that of L-arginine, leading to a redox imbalance that activates nuclear factor kappaB exacerbating inflammation and NO mediated cytotoxicity. Physical exercise is capable of inducing changes in the pattern of cytokine production and release towards type 2 class and to normalize the glutamine supply to the circulation, which reduces the need for glutamate, whose metabolic fate may be restored in the direction of GSH synthesis and antioxidant defence. Also, the 70-kDa heat shock protein (hsp70), which is immunoregulatory, may modulate exercise-induced anti-inflammation. In this work, we envisage how exercise can intervene in the mechanisms involved in the autoimmune process against beta-cells and how novel therapeutic approaches may be inferred from these observations. PMID- 18383560 TI - Missing item imputation for quality-of-life instruments with application to asthma quality-of-life questionnaires. AB - There has been increasing use of quality-of-life (QoL) instruments in drug development. Missing item values often occur in QoL data. A common approach to solve this problem is to impute the missing values before scoring. Several imputation procedures, such as imputing with the most correlated item and imputing with a row/column model or an item response model, have been proposed. We examine these procedures using data from two clinical trials, in which the original asthma quality-of-life questionnaire (AQLQ) and the miniAQLQ were used. We propose two modifications to existing procedures: truncating the imputed values to eliminate outliers and using the proportional odds model as the item response model for imputation. We also propose a novel imputation method based on a semi-parametric beta regression so that the imputed value is always in the correct range and illustrate how this approach can easily be implemented in commonly used statistical software. To compare these approaches, we deleted 5% of item values in the data according to three different missingness mechanisms, imputed them using these approaches and compared the imputed values with the true values. Our comparison showed that the row/column-model-based imputation with truncation generally performed better, whereas our new approach had better performance under a number scenarios. PMID- 18383561 TI - Common adverse events associated with an SSRI: meta-analysis of early paroxetine data. AB - PURPOSE: We wanted to determine to what extent adverse drug effects associated with a selective serotonin reuptake inhibitor (SSRI) were known but not assessed before application for registration of paroxetine. METHODS: With a special permit from the Norwegian Ministry of Health, we obtained reports from 82 clinical trials presented by the paroxetine license holder in 1989. There were 17 double blind, placebo controlled clinical trials with parallel design with 903 patients on paroxetine and 592 on placebo. Altogether 32 adverse effects showed a risk difference (RD) between paroxetine and control groups of more than 0.5%. We did a meta-analysis for each of these adverse effects. We then compared the outcome with the frequencies stated in the Summary of Product Characteristics (SPC) at the time of registration and those reported in the current SPC. RESULTS: At the time of registration 19 of the adverse effects were statistically significant. Only eight of these adverse effects were listed as being common in the first SPC from 1989. Five out of the nineteen adverse effects are not mentioned in the current SPC. Among them are headache with RD 5.4%, decreased libido RD 2.6%, nervousness RD 2.0% and paresthesia RD 1.7%. CONCLUSIONS: Frequently occurring adverse reactions that are included in today's SPC for paroxetine were evident and documented already in the early studies accompanying the application for marketing authorization in 1989. Some other adverse effects observed then are still not mentioned in the SPC of today. Meta-analyses of adverse effects should be mandatory at the stage of first registration of a drug. PMID- 18383562 TI - For the majority of patients with diabetes blood pressure and lipid management is not in line with recommendations. Results from a large population-based cohort in Germany. AB - OBJECTIVE: To evaluate the management of cardiovascular risk factors in a population-based cohort of adults with diabetes mellitus in Germany. DESIGN AND MEASUREMENTS: For the 9953 participants (50-74 years of age) of the ESTHER study, diabetes mellitus, hypertension, dyslipidemia and the respective medication were documented at baseline by their primary care physician. Blood pressure was taken and lipid levels were determined from a blood sample drawn at recruitment. Lifestyle factors (smoking, BMI, physical activity) were documented by the participants. Prevalence of pharmacotherapy for and control of hypertension and dyslipidemia among diabetic patients with these diagnoses were assessed and determinants were evaluated by multiple logistic regression. RESULTS: Diabetes mellitus was present in 1375 participants (14.2%). Almost 78% of diabetic participants had physician diagnosed hypertension, 86.0% received pharmacotherapy, but only 12.8% of those with anti-hypertensive medication achieved blood pressure levels below 130/85 mmHg. Obese participants were more likely to receive anti-hypertensive pharmacotherapy than non-obese ones: adjusted odds ratio (OR, 95% confidence interval (CI)): 3.58 (1.86-6.87). Gender had no influence on anti-hypertensive pharmacotherapy. Older diabetic patients with hypertension were less likely to have sufficient blood pressure control than younger ones. Dyslipidemia was documented in 50% of diabetic patients. Diabetic participants with coronary heart disease or hypertension were more likely to receive lipid-lowering pharmacotherapy than those without these conditions (adjusted OR 1.85 (95%CI 1.19-2.89) and 2.59 (95%CI 1.41-4.74), respectively). CONCLUSIONS: For most elderly with diabetes cardiovascular risk factor management continues to be not in line with recommendations. PMID- 18383563 TI - Toxicological and biochemical response to azinphos-methyl in Cydia pomonella L. (Lepidoptera: Tortricidae) among orchards from the Argentinian Patagonia. AB - BACKGROUND: Azinphos-methyl is the main insecticide used to control codling moth on apple and pears in Northern Patagonia. The aim of this study was to evaluate the toxicological and biochemical response of diapausing larvae of codling moth in orchards subjected to different insecticide selection pressure. RESULTS: Dose mortality assays with azinphos-methyl in diapausing larvae of Cydia pomonella L. showed significant differences between the LD(95) from a population collected in one untreated orchard (2.52 microg moth(-1)) compared with that in a laboratory susceptible population (0.33 microg moth(-1)). Toxicity to azinphos-methyl in field populations of diapausing larvae collected during 2003-2005 was evaluated by topical application of a discriminating dose (2.5 microg moth(-1)) that was obtained from larvae collected in the untreated orchard (field reference strain). Significantly lower mortality (37.71-84.21%) was observed in three out of eight field populations compared with that in the field reference strain. Most of the field populations showed higher esterase activity than that determined in both the laboratory susceptible and the field reference strains. Moreover, there was a high association between esterase activity and mortality (R(2)=0.64) among the field populations. On the other hand, a poor correlation was observed between glutathione S-transferase activity and mortality (R(2)=0.33) among larvae collected from different orchards. CONCLUSIONS: All the field populations evaluated exhibited some degree of azinphos-methyl tolerance in relation to the laboratory susceptible strain. Biochemical results demonstrated that esterases are at least one of the principal mechanisms involved in tolerance to this insecticide. PMID- 18383564 TI - Angiotensin II induces endothelial cell senescence via the activation of mitogen activated protein kinases. AB - Vascular endothelial cells have a finite cell lifespan and eventually enter an irreversible growth arrest, cellular senescence. The functional changes associated with cellular senescence are thought to contribute to human aging and age-related cardiovascular disorders, for example, atherosclerosis. Angiotensin II (Ang II), a principal effector of the renin-angiotensin system (RAS), an important signaling molecule involved in atherogenic stimuli, is known to promote aging and cellular senescence. In the present study, induction of Ang II promoted a growth arrest with phenotypic characteristics of cell senescence, such as enlarged cell shapes, increased senescence-associated beta-galactosidase (SA-beta gal) positive staining cells, and depressed cell proliferation. Ang II drastically decreased the expression level of Bcl-2, in part via the activation of extracellular signal-regulated kinase (ERK). Our results suggest that Ang II can induce HUVEC senescence; one of its molecular mechanisms is a probability that the mitogen-activated protein kinase (MAPK) signal pathway is involved in the process of pathological and physiological senescence of endothelial cells as well as vascular aging. PMID- 18383565 TI - Evaluation of lure dispensers for fruit fly surveillance in New Zealand. AB - BACKGROUND: Fruit flies (Diptera: Tephritidae) represent a major biosecurity threat to the horticulture sector of New Zealand, which is entirely free of these invasive pests. A nationwide surveillance programme is conducted to ensure any incursion is detected as early as possible. A review of the lure dispensers used is reported here. RESULTS: Lure dispenser emission trials found that the currently used lure plugs release lure more slowly under New Zealand subtropical to temperate climates than wafer dispensers. Subsequent trapping experiments at high altitude in Hawaii (as a mimic of New Zealand meteorological and expected fruit fly ecological conditions) compared Lynfield traps baited with the existing lure plug dispensers and newer wafer dispensers. Catches of wild Oriental fruit flies, Bactrocera dorsalis (Hendel), were 9.5-fold higher with methyl eugenol wafers than with the plugs. Recaptures of sterile melon flies, Bactrocera cucurbitae (Coquillet), were 2.6-fold higher with cuelure wafers than with the plugs. Recaptures of sterile Mediterranean fruit flies, Ceratitis capitata Weid., were not significantly higher with trimedlure wafers than with the plugs. CONCLUSIONS: Release rate and trapping experiments found new lure dispensers differed in release rate characteristics from existing dispensers under temperate and subtropical conditions, and indicated some potential for improvement in surveillance efficacy. PMID- 18383566 TI - Lipopolysaccharide-induced monocyte chemotactic protein-1 is enhanced by suppression of nitric oxide production, which depends on poor CD14 expression on the surface of skeletal muscle. AB - It is known that lipopolysaccharide (LPS)-induced monocyte chemotactic protein (MCP)-1 secretion from tissues recruits monocytes from the circulation, but the mechanism of the LPS-induced MCP-1 production in skeletal muscle is largely unexplained. To clarify the effect of LPS on MCP-1 production in skeletal muscle cells, C2C12 cells from a mouse skeletal muscle cell line, and RAW 264.7 cells from a mouse macrophage cell line, were used to assess production of LPS-induced MCP-1, nitric oxide (NO) and interferon (IFN)-beta. In addition, we evaluated inducible NO synthases (iNOS) mRNA expression using RT-PCR, and cell surface expression of CD14 and toll-like receptor (TLR) 4 using flow cytometry. In C2C12 cells, LPS stimulation increased MCP-1 production (p < 0.01), but combined treatment with LPS and NO inducer, diethylammonium (Z)-1-(N,N-diethylamino) diazen-1-ium-1,2-diolate (NONOate), significantly inhibited its production (p < 0.01). LPS stimulation neither induced production of NO nor of IFN-beta, which is an NO inducer. Recombinant IFN-beta stimulation, on the other hand, enhanced LPS induced NO production (p < 0.01). Interestingly, we found that surface expression of CD14, which regulates IFN-beta production, in C2C12 cells was much lower than that in RAW 264.7 cells, although TLR4 expression on C2C12 cells was similar to that on RAW 264.7 cells. These data suggest that the reduced NO production in response to LPS may depend on low expression of CD14 on the cell surface of skeletal muscle, and that it may enhance LPS-induced MCP-1 production. Together, these functions of skeletal muscle could decrease the risk of bacterial infection by recruitment of monocytes. PMID- 18383567 TI - Coreduction colloidal synthesis of III-V nanocrystals: the case of InP. PMID- 18383568 TI - Rh(I)-catalyzed hydration of organonitriles under ambient conditions. PMID- 18383569 TI - Syntheses and comparison of 2,6-di-O-methyl celluloses from natural and synthetic celluloses. AB - 2,6-Di-O-methylcellulose was prepared from natural and synthetic celluloses. Natural cellulose was converted to 2,6-di-O-thexyldimethylsilylcellulose, then to 3-mono-O-allyl-2,6-di-O-methylcellulose, and finally into 2,6-di-O methylcellulose. Alternatively, 2,6 di-O-methylcellulose was synthesized from the synthetic cellulose derivative 3-mono-O-benzyl-2,6-di-O-pivaloylcellulose by depivaloylation and methylation to give 3-mono-O-benzyl-2,6-di-O-methylcellulose, which was debenzylated to yield the dimethyl ether. Both types of 2,6-di-O methylcellulose are insoluble in water and common organic solvents. The structures of all cellulose derivatives were determined by NMR. PMID- 18383570 TI - AFM imaging of RGD presenting synthetic extracellular matrix using gold nanoparticles. AB - Several high-resolution imaging techniques such as FESEM, TEM and AFM are compared with respect to their application on alginate hydrogels, a widely used polysaccharide biomaterial. A new AFM method applicable to RGD peptides covalently conjugated to alginate hydrogels is described. High-resolution images of RGD adhesion ligand distribution were obtained by labeling biotinylated RGD peptides with streptavidin-labeled gold nanoparticles. This method may broadly provide a useful tool for sECM characterization and design for tissue regeneration strategies. PMID- 18383572 TI - Surface functionalization of porous resorbable scaffolds by covalent grafting. AB - Resorbable porous scaffold discs and solid films were prepared from poly[(1,5 dioxepan-2-one)-co-(L-lactide)] and poly[(epsilon-caprolactone)-co-(L-lactide)]. The surfaces of the scaffolds were functionalized to increase their hydrophilicity. A total of 90 samples were prepared to cover all important combinations of experimental and material factors, and all experimental data were fitted by a partial least square model. As a result of grafting, the porous discs and solid films exhibited a tremendous increase in wettability. The functionalized discs were hygroscopic so that water was instantly absorbed and thoroughly wet the substrates. PMID- 18383571 TI - New insights into polylactide biodegradation from molecular ecological techniques. AB - Molecular ecological techniques for direct identification of microbes involved in PLA degradation under aerobic composting conditions are described. Gene sequences from genera Paecilomyces, Thermomonospora, and Thermopolyspora were most abundant in the compost samples. Members of these phylogenetic lineages are therefore likely to play an important role in PLA degradation. The use of molecular ecological techniques to design cultivation strategies may also provide a new tool for identification and investigation of biodegradation mechanisms and for future development of efficient biological treatment or recycling processes for PLA and other biodegradable polymers. PMID- 18383573 TI - Solid-state microwave-arcing-induced formation and surface functionalization of core/shell metal/carbon nanoparticles. PMID- 18383574 TI - Following the autonomous movement of silica microparticles using fluorescence microscopy. PMID- 18383575 TI - Direct soft-chemical synthesis of chalcogen-doped manganese oxide 1D nanostructures: influence of chalcogen doping on electrode performance. AB - We have developed a direct nonhydrothermal route to nanostructured chalcogen doped manganese oxides; K(x)MnO(2)Q(y) (Q = S, Se, and Te). According to combinative diffraction and microscopic analyses, the S- and Se-doped manganese oxides exhibit 1D nanowire-type morphology with layered delta-MnO(2)- and alpha MnO(2)-structures, respectively, whereas the Te-doped compound consists of 3D nanospheres that are amorphous according to X-ray diffraction. X-ray absorption and X-ray photoelectron spectroscopy analyses clearly demonstrate that the doped chalcogen ions exist in the form of hexavalent chalcogenate clusters mainly on the sample surface or grain boundary. According to electrochemical and ex situ X ray absorption spectroscopy investigations, the Se-doped manganate nanowires show higher structural stability and better electrode performance with excellent rate characteristics compared to the S-/Te-doped and undoped manganate nanostructures. This is attributed to the presence of chemically stable SeO4(2-) species, leading to enhanced stability of the manganate lattice through the prevention of structural deformation during cycling and/or to the improvement of Li(+) ion transport through the maintenance of intercrystallite voids. Based on the present experimental findings, we are able to conclude that the present one-pot soft chemical route with chalcogen dopants can provide a simple method not only to economically synthesize 1D nanostructured manganese oxides but also to finely control their electrode performance, crystal structure and morphology, and lattice stability. PMID- 18383576 TI - Light-activated nanoimpeller-controlled drug release in cancer cells. PMID- 18383577 TI - Colorimetric response of peptide-functionalized gold nanoparticles to metal ions. PMID- 18383578 TI - Polyaniline-coated Fe3O4 nanoparticle-carbon-nanotube composite and its application in electrochemical biosensing. PMID- 18383579 TI - Autonomous loading, transport, and unloading of specified cargoes by using DNA hybridization and biological motor-based motility. PMID- 18383580 TI - DNA-carbon nanotube conjugates prepared by a versatile method using streptavidin biotin recognition. PMID- 18383581 TI - p16INK4a downregulation is involved in immortalization of primary human prostate epithelial cells induced by telomerase. AB - Prostate cancer is a major cause of cancer death among male population. Therefore, development of appropriate model systems is critical for understanding the molecular basis of prostate cancer progression. In this study, introduction of human telomerase (hTERT) into normal human prostate epithelial cells (PrECs) renders them higher telomerase activity, elongated telomere length and an extended proliferative lifespan. The immortal mass culture of PrEC-hTERT cell line with stabilized telomere length has been established using hTERT transfection. However, activation of hTERT alone appears to be insufficient for immortalization of PrEC cells because methylation of p16(INK4a) promoter has been found to be involved in the immortalization process. p53 was functionally intact and no mutations of p53 gene were identified in the immortalized PrECs. In addition, the immortal PrECs show a near diploid complement of chromosomes albeit a few reciprocal and non-reciprocal translocations are identified. They are anchorage dependent and do not form tumors in immunosuppressed host animals. Therefore, premalignantly transformed human PrECs provide a valuable model for prostate cancer research. PMID- 18383582 TI - Potentially functional polymorphisms of the vascular endothelial growth factor gene and risk of gastric cancer. AB - Vascular endothelial growth factor (VEGF), the key mediator of angiogenesis, plays an important role in the development of different kind of tumors, including gastric cancer (GC). The aim of this study is to test the hypothesis that genetic variants of VEGF are associated with risk of GC. We genotyped four potentially functional polymorphisms (-2578C > A, -1498T > C, -634G > C, and +936C > T) of the VEGF gene in a population-based case-control study of 540 GC cases and 561 frequency-matched cancer-free controls in a high risk Chinese population. We found that none of the four polymorphisms or their haplotypes achieved significant difference in their distributions between GC cases and controls. Multiple logistic regression analyses revealed that GC risk was not significantly associated with the variant genotypes of the four VEGF polymorphisms as compared with their wild-type genotypes. In conclusion, our data did not support a significant association between VEGF SNPs and the risk of GC. PMID- 18383583 TI - Probing the hydrophobic pocket of the active site in the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) by variable stereoselective alkane hydroxylation and olefin epoxidation. AB - pMMO from M. capsulatus (Bath) oxidizes straight-chain C1-C5 alkanes and alkenes to form their corresponding 2-alcohols and epoxides. According to experiments performed with cryptically chiral ethane and D,L-[2-(2)H(1),3-(2)H(1)]butane, the reactions proceed through the concerted O-atom insertion mechanism. However, when propene and but-1-ene are used as epoxidation substrates, the enantiomeric excesses (ees) of the enzymatic products are only 18 and 37 %, respectively. This relatively poor stereoselectivity in the enzymatic epoxidation presumably reflects low stereochemical differentiation between the re and si faces in the hydrophobic pocket of the active site. Further insights into the reaction mechanism are now provided by studies on trans-but-2-ene, which reveal only the D,L-2,3-dimethyloxirane products, and on cis-but-2-ene, which yield only the meso product. These observations indicate that the enzymatic epoxidation indeed proceeds through electrophilic syn addition. To achieve better facial selectivity, we have also used 3,3,3-trifluoroprop-1-ene as the substrate. The products obtained are 90 % (2S)-oxirane. When 1,1,1-trifluoropropane is the substrate, the hydroxylation at the 2-carbon exhibits an inverse chiral selectivity relative to that seen with normal butane, if we consider the size of the CF(3) group in the fluorinated propane to be comparable to one of the ethyl groups in butane. These experiments are beginning to delineate the factors that influence the orientations of various substrates in the hydrophobic cavity of the active site in the enzyme. PMID- 18383584 TI - The mineralisation of fresh and humified soil organic matter by the soil microbial biomass. AB - Soil organic matter comprises all dead plant and animal residues, from the most recent inputs to the most intensively humified. We have found that traces of fresh substrates at microg g(-1) soil concentrations (termed 'trigger molecules') activate the biomass to expend more energy than is contained in the original 'trigger molecules'. In contrast, we suggest that the rate limiting step in soil organic matter mineralisation is independent of microbial activity, but is governed by abiological processes (which we term the Regulatory Gate theory). These two findings have important implications for our understanding of carbon mineralisation in soil, a fundamental process in the sequestration of soil organic matter. PMID- 18383585 TI - Chest X-ray quiz. Constipation and pneumothorax. PMID- 18383586 TI - A young woman with slowly-progressive generalized dystonia. PMID- 18383587 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383588 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383589 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383590 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383591 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383592 TI - Novel human pathological mutations. Gene symbol: CFTR. Disease: cystic fibrosis. PMID- 18383593 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383594 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383595 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383596 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383597 TI - Novel human pathological mutations. Gene symbol: KRIT1. Disease: cerebral cavernous malformation. PMID- 18383598 TI - Novel human pathological mutations. Gene symbol: MYOC. Disease: primary open angle glaucoma. PMID- 18383599 TI - Novel human pathological mutations. Gene symbol: SLC3A1. Disease: cystinuria. PMID- 18383600 TI - Novel human pathological mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18383601 TI - Methicillin-resistant Staphylococcus aureus in hospitals: time for a culture change. PMID- 18383602 TI - Novel human pathological mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18383603 TI - Novel human pathological mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18383604 TI - Novel human pathological mutations. Gene symbol: UROD. Disease: porphyria, cutaneous. PMID- 18383605 TI - Novel human pathological mutations. Gene symbol: UROD. Disease: porphyria, cutaneous. PMID- 18383606 TI - Novel human pathological mutations. Gene symbol: RDS. Disease: macular dystrophy. PMID- 18383608 TI - Novel human pathological mutations. Gene symbol: TYR. Disease: albinism, oculocutaneous 1. PMID- 18383607 TI - [Per- and subtrochanteric fractures]. PMID- 18383609 TI - Novel human pathological mutations. Gene symbol: TYR. Disease: tyrosinase deficiency. PMID- 18383610 TI - Novel human pathological mutations. Gene symbol: TYR. Disease: tyrosinase deficiency. PMID- 18383611 TI - Novel human pathological mutations. Gene symbol: TYR. Disease: tyrosinase deficiency. PMID- 18383612 TI - Novel human pathological mutations. Gene symbol: PMP22. Disease: Charcot-Marie Tooth disease 1. PMID- 18383613 TI - Novel human pathological mutations. Gene symbol: GJB1. Disease: Charcot-Marie Tooth disease 1. PMID- 18383614 TI - An ounce of prevention. PMID- 18383615 TI - Multiple pathways for regulation of sigmaS (RpoS) stability in Escherichia coli via the action of multiple anti-adaptors. AB - SigmaS, the stationary phase sigma factor of Escherichia coli and Salmonella, is regulated at multiple levels. The sigmaS protein is unstable during exponential growth and is stabilized during stationary phase and after various stress treatments. Degradation requires both the ClpXP protease and the adaptor RssB. The small antiadaptor protein IraP is made in response to phosphate starvation and interacts with RssB, causing sigmaS stabilization under this stress condition. IraP is essential for sigmaS stabilization in some but not all starvation conditions, suggesting the existence of other anti-adaptor proteins. We report here the identification of new regulators of sigmaS stability, important under other stress conditions. IraM (inhibitor of RssB activity during Magnesium starvation) and IraD (inhibitor of RssB activity after DNA damage) inhibit sigmaS proteolysis both in vivo and in vitro. Our results reveal that multiple anti-adaptor proteins allow the regulation of sigmaS stability through the regulation of RssB activity under a variety of stress conditions. PMID- 18383616 TI - FliZ, a flagellar regulator, is at the crossroads between motility, haemolysin expression and virulence in the insect pathogenic bacterium Xenorhabdus. AB - There is a complex interplay between the regulation of flagellar motility and the expression of virulence factors in many bacterial pathogens. We investigated the role of FliZ in the regulation of flagellar and virulence genes in Xenorhabdus nematophila, an insect pathogen. The fliZ gene is the second gene in the fliAZ operon in X. nematophila. In vivo transcription analysis revealed a positive feedback loop of fliAZ transcription in which FliZ activates flhDC, the master operon of flagellar regulon in X. nematophila, leading to an increased transcription of the FlhDC-dependent promoter of fliAZ. We also showed that fliAZ and flhDC mutants lacked motility, had no haemolysin or Tween lipase activity and displayed an attenuated virulence phenotype in insects. Lipase activity is controlled by FliA, whereas haemolysin production and full virulence phenotype have been reported to be FliZ-dependent. Transcriptional analysis revealed that FliZ directly controlled expression of the xhlBA and xaxAB operons, which encode haemolysins from the two-partner secretion system and the binary XaxAB toxin family respectively. We suggest that this regulatory pathway may also occur in other pathogenic enterobacteria with genes encoding members of these two growing families of haemolysins. PMID- 18383617 TI - [Relationship of the specifications and informative value of digital X-ray study in pulmonary tuberculosis]. AB - Digital fluorographs used during mass chest screenings have different specifications that affect the diagnostic value of an obtained image. Visualization using on various apparatuses was compared in the context of early detection of tuberculosis of the most important skialogic signs of tuberculosis. The findings make it possible to substantively appraise the capacities of digital X-ray systems in detecting the early signs of tuberculosis and monitoring its process over time. PMID- 18383619 TI - [The historical perspective of the sentinel lymph node concept]. PMID- 18383618 TI - [Validity of ultrasound fetometry in the diagnosis of a large fetus during screening]. AB - The purpose of the study was to assess the validity of fetometry in the diagnosis of a large fetus during screening. Eight hundred and sixty-five case histories of large fetal labors were analyzed, by determining the assumed fetal mass by ultrasound fetometry before labor. The studies have demonstrated that the validity of fetometry in the diagnosis of a large fetus was not greater than 73%. This was caused by methodical errors, the specific features of different procedures for calculating the assumed fetal mass, and the heterogeneity of anthropometric parameters of large fetuses. CONCLUSIONS: it is necessary to study additional possibilities of enhancing the validity of fetometry in the determination of a large fetus during screening and to continue a search for optimal methods of calculating the assumed fetal mass. PMID- 18383620 TI - Effects of lipid-lowering therapy on coronary heart disease in older patients: the SAGE study. PMID- 18383621 TI - Rosuvastatin and carotid intima-media thickness: the METEOR trial. PMID- 18383622 TI - Torcetrapib and the progression of coronary atherosclerosis. PMID- 18383623 TI - Beyond PrP res type 1/type 2 dichotomy in Creutzfeldt-Jakob disease. AB - Sporadic Creutzfeldt-Jakob disease (sCJD) cases are currently subclassified according to the methionine/valine polymorphism at codon 129 of the PRNP gene and the proteinase K (PK) digested abnormal prion protein (PrPres)identified on Western blotting (type 1 or type 2). These biochemically distinct PrPres types have been considered to represent potential distinct prion strains. However, since cases of CJD show co-occurrence of type 1 and type 2 PrPres in the brain, the basis of this classification system and its relationship to agent strain are under discussion. Different brain are as from 41 sCJD and 12 iatrogenic CJD (iCJD) cases were investigated, using Western blotting for PrPres and two other biochemical assays reflecting the behaviour of the disease-associated form of the prion protein (PrPSc) under variable PK digestion conditions. In 30% of cases, both type 1 and type 2 PrPres were identified. Despite this, the other two biochemical assays found that PrPSc from an individual patient demonstrated uniform biochemical properties. Moreover, in sCJD, four distinct biochemical PrPSc subgroups were identified that correlated with the current sCJD clinico pathological classification. In iCJD, four similar biochemical clusters were observed, but these did not correlate to any particular PRNP 129 polymorphism or western blot PrPres pattern. The identification of four different PrPSc biochemical subgroups in sCJD and iCJD, irrespective of the PRNP polymorphism at codon 129 and the PrPres isoform provides an alternative biochemical definition of PrPSc diversity and new insight in the perception of Human TSE agents variability. PMID- 18383624 TI - The time is right: proteome biology of stem cells. AB - In stem cell biology, there is a growing need for advanced technologies that may help to unravel the molecular mechanisms of self-renewal and differentiation. Proteomics, the comprehensive analysis of proteins, is such an emerging technique. To facilitate interactions between specialists in proteomics and stem cell biology,a new initiative has been undertaken, supported by the Human Proteome Organization (HUPO) and the International Society for Stem Cell Research (ISSCR). Here we present the Proteome Biology of Stem Cells Initiative (PBSCI) and report on its goals and future activities. PMID- 18383625 TI - Editor's notes. PMID- 18383628 TI - Nuclear cardiology in the literature. PMID- 18383627 TI - Ethical standards for human-to-animal chimera experiments in stem cell research. PMID- 18383629 TI - [A multidimensional description and validation of two subtypes in the field of endogenous and melancholic depression]. AB - BACKGROUND: It should be possible to improve the diagnosis of endogenous or melancholic depression by applying Jaspers' multidimensional model of clinical symptoms, by considering the role of vasopressinergic activity in dysregulated stress reactions and by referring to Cloninger's personality model. METHOD: The strategy of Robins and Guze is used to develop diagnostic concepts. Melancholic depression according to DSM-IV criteria is converted into a mixture of basic symptom dimensions and this mixture is investigated for its possible links with familial depression, time needed for complete recovery, plasma vasopressin concentration, correlated plasma vasopressin- and cortisolconcentrations, and features of temperament and character. RESULTS: Two subtypes were found. One is a highly anxious-retarded subtype, with slow recovery, a correlation between vasopressin and cortisol, low pre-morbid self-directedness and a positive family history of depression; the other is a subtype with an above-normal plasma vasopressin concentration, an anxious-retarded phenotype without an intensity threshold, correlated vasopressin- and cortisol-concentration, low pre-morbid cooperativeness and a positive family history of depression. CONCLUSION: A multidimensional description of clinical symptoms and personality makes it possible to integrate pathophysiological aspects into new diagnostic concepts. PMID- 18383626 TI - Chlamydial entry involves TARP binding of guanine nucleotide exchange factors. AB - Chlamydia trachomatis attachment to cells induces the secretion of the elementary body-associated protein TARP (Translocated Actin Recruiting Protein). TARP crosses the plasma membrane where it is immediately phosphorylated at tyrosine residues by unknown host kinases. The Rac GTPase is also activated, resulting in WAVE2 and Arp2/3-dependent recruitment of actin to the sites of chlamydia attachment. We show that TARP participates directly in chlamydial invasion activating the Rac-dependent signaling cascade to recruit actin. TARP functions by binding two distinct Rac guanine nucleotide exchange factors (GEFs), Sos1 and Vav2, in a phosphotyrosine-dependent manner. The tyrosine phosphorylation profile of the sequence YEPISTENIYESI within TARP, as well as the transient activation of the phosphatidylinositol 3-kinase (PI3-K), appears to determine which GEF is utilized to activate Rac. The first and second tyrosine residues, when phosphorylated, are utilized by the Sos1/Abi1/Eps8 and Vav2, respectively, with the latter requiring the lipid phosphatidylinositol 3,4,5-triphosphate. Depletion of these critical signaling molecules by siRNA resulted in inhibition of chlamydial invasion to varying degrees, owing to a possible functional redundancy of the two pathways. Collectively, these data implicate TARP in signaling to the actin cytoskeleton remodeling machinery, demonstrating a mechanism by which C.trachomatis invades non-phagocytic cells. PMID- 18383630 TI - Diterpenoids from Copaifera reticulata Ducke with larvicidal activity against Aedes aegypti (L.) (Diptera, Culicidae). AB - The objective of this study was to evaluate the larvicidal activity of diterpenoids obtained from the oil-resin of Copaifera reticulata against Aedes aegypti larvae, the principal vector of dengue and urban yellow fever. Four diterpenes were obtained from oil-resin extraction with organic solvents and subsequent chromatographic and spectroscopic procedures allowed to isolation and identification of these compounds as 3-beta-acetoxylabdan-8(17)-13-dien-15-oic acid (1), alepterolic acid (2), 3-beta-hidroxylabdan-8(17)-en-15-oic acid (3), and ent-agatic acid (4). Each compound was previously dissolved in dimethylsulphoxide, and distilled water was added to obtain the desired concentrations. Twenty larvae of third instars were placed into plastic beckers, containing the solution test (25 mL), in a five repetitions scheme, and their mortality, indicated by torpor and darkening of the cephalic capsule, was recorded after 48 h. Probit analyses were used to determine lethal concentrations (LC50 and LC90) and their respective 95% confidence intervals. This study showed that only diterpenoids 1 and 2 exhibited larvicidal properties with LC50 of 0.8 ppm and 87.3 ppm, respectively, revealing the former as the most toxic compound against third instars of Ae. aegypti. Therefore, this compound seems to be an interesting source for new metabolite to be exploited. PMID- 18383631 TI - Systematics of Euromediterranean Silene (Caryophyllaceae): evidence from a phylogenetic analysis using ITS sequences. AB - The generic boundaries within the sub-family Silenoideae (Caryophyllaceae), and especially within the complex Silene-Lychnis-Melandrium have long been controversial. These 3 genes are now grouped in a single genus: Silene. In order to test the monophyly of this genus and to clarify intra-generic phylogenetic relationships, we compared ITS sequences of 22 species of Euromediterranean Silene chosen throughout the classification, and 4 putative outgroup species. The genus Silene in its current circumscription formed a well-supported monophyletic group. Lychnis and Melandrium species are scattered within the genus Silene. The monotypic genus Cucubalus also fell within the Silene clade. PMID- 18383632 TI - [Kinetic analysis of historical processes: the model case of the health care system]. PMID- 18383633 TI - [Enforcement of prion research and prevalence of prion disease]. PMID- 18383634 TI - Life insurance, the medical examination and cultural values. PMID- 18383635 TI - [The will to quantify: conceptions of measurement of medical practice]. PMID- 18383636 TI - Hepatitis B vaccines (continued): risks of systemic adverse reactions to hepatitis B vaccines appear to be very rare. PMID- 18383637 TI - Haemorrhage due to Ginkgo biloba? AB - (1) About 20 detailed reports of haemorrhage (usually cerebral, ocular, or postsurgical) in patients using Gingko biloba extracts have been published. One third of these patients were also taking drugs that increase the risk of bleeding (anticoagulants or antiplatelet drugs). (2) Some substances contained in Gingko biloba have been shown to have an antiplatelet effect. (3) In practice, patients with risk factors for bleeding (anticoagulant or antiplatelet treatment, surgery, etc.) should avoid using Gingko biloba extracts. PMID- 18383638 TI - Fluoroquinolones: psychiatric adverse effects. AB - (1) In France, between 1985 and 2002, there were 590 reports of psychiatric adverse effects in patients receiving fluoroquinolones. These mainly included cases of confusion, hallucinations, agitation, delirium, insomnia and drowsiness. Elderly patients appear to be at greatest risk. (2) The reports implicated all available fluoroquinolones and, in most cases, oral intake. (3) Fluoroquinolone dose regimens should be reduced in cases of renal failure. The Cockcroft formula is used to derive creatinine clearance from plasma creatinine levels. PMID- 18383639 TI - Linking agricultural biodiversity and food security: the valuable role of agrobiodiversity for sustainable agriculture. PMID- 18383640 TI - The collective mind: trauma and shell-shock in twentieth-century Russia. PMID- 18383641 TI - Dental school vacant budgeted faculty positions, academic years 2005-06 and 2006 07. AB - The annual turnover of dental school faculty creates a varying number of vacant budgeted positions at any given time. The American Dental Education Association (ADEA) conducts an annual survey to determine the status and characteristics of these vacant faculty positions. In addition, ADEA conducts an annual survey of dental educators to maintain a database on the size and characteristics of dental school faculty, including data on the distribution of full-time, part-time, and volunteer faculty, reasons for faculty separations, and sources of new faculty. The number of vacant budgeted faculty positions within U.S. dental schools increased throughout the 1990s, with a peak of 358 positions in 2000. Following this peak, the number of vacancies declined, falling to 275 in 2004-05. Since that time, there has been a rapid increase in the number of estimated vacancies, reaching 417 in 2005-06, then falling slightly to 406 in 2006-07. The 2005-06 and 2006-07 faculty vacancies surveys explore these increases, along with information relevant to trends in the faculty workforce, factors influencing faculty vacancies, and the impact of vacant positions on dental schools. PMID- 18383642 TI - Fateful memories: industrialized war and traumatic neuroses. PMID- 18383643 TI - Collingwood on re-enactment and the identity of thought. PMID- 18383644 TI - Is a prescriptive or an open referral related to physical therapy outcomes in patients with lumbar spine-related problems? AB - STUDY DESIGN: Retrospective review of medical records. OBJECTIVES: To describe characteristics of patients with lumbar spine dysfunction, and to compare functional outcome and number of visits to physical therapy according to type of physician referral. BACKGROUND: The type of referral, characterized as prescriptive or open, has been associated with the perceived amount of supervision that is required for the provision of physical therapy care. The rationale for prescriptive referrals is not consistent with autonomous physical therapy practice, and may be deemed unnecessary if such referrals are associated with equivalent outcomes. METHODS AND MEASURES: Medical records of patients treated within a rehabilitation provider network between October 2002 and December 2003 were reviewed retrospectively for administrative and clinical variables. Subjects were selected if they completed the Roland Morris Questionnaire (RMQ) on admission and discharge from physical therapy care. Associations between referral type, discharge RMQ scores, and number of visits were determined using independent t tests and were further examined using stepwise multiple regression analysis. RESULTS: Ninety-six records met inclusion criteria, of which 54 (56.2%) had open referrals and 41 (43.8%) had prescriptive referrals. Type of referral was not associated with number of visits in bivariate or in multivariate analysis. Prescriptive referrals were associated with higher discharge RMQ scores, representing greater disability, in bivariate analysis (t test, P = .03); however, this association was attenuated in multivariate analyses after adjustment for physician status as primary care practitioner or specialist. CONCLUSION: Prescriptive referrals were not associated with enhanced outcomes of physical therapy care. PMID- 18383645 TI - Centralization: prevalence and effect on treatment outcomes using a standardized operational definition and measurement method. AB - STUDY DESIGN: Retrospective, observational cohort design. OBJECTIVES: Purpose 1 was to determine the association between age, symptom chronicity, and prevalence of centralization in a sample of patients with nonserious cervical or lumbar spinal syndromes referred to a hospital-based outpatient rehabilitation clinic. Purpose 2 was to examine if classifying these patients at intake to centralization or noncentralization predicts functional status, pain intensity, and number of treatment visits at discharge from rehabilitation. Purpose 3 was to compare clinically meaningful changes in functional status and pain intensity between patients subgrouped by centralization and noncentralization. BACKGROUND: Variations in operational definitions and measurements used to identify centralization affect patient classification, contribute to variation in reported prevalence rates, and influence treatment strategy and outcome interpretation. Investigating a standardized operational definition and measurement method for centralization may reduce practice and outcomes variation. METHODS AND MEASURES: Adults (n=418) with cervical or low back syndromes (mean +/- SD age, 58 +/- 17 years; range, 19-91 years; 33% male; 76% lumbar symptoms; 53% chronic symptoms) were assessed. Therapists classified patients using a standardized operational definition and method for centralization during initial evaluation. Prevalence rates were calculated for centralization by age and acuity. Multivariate models were used to assess discharge functional status, pain intensity, and visits while controlling important variables. Percentage of patients subgrouped by centralization and noncentralization achieving minimal clinically important differences (MCID) in functional status and pain intensity was assessed. RESULTS: Overall prevalence rate for centralization was 17%, but increased for patients who were younger and reported acute symptoms regardless of body part. For patients with lumbar syndromes, noncentralization was associated with lower discharge functional status and more pain, but not associated with number of visits compared to patients classified as centralization. For patients with cervical syndromes, noncentralization was associated with more pain but not associated with functional status or number of visits compared to patients classified as centralization. Pain pattern classification affected percentage of patients with lumbar and cervical impairment achieving MCID. CONCLUSION: Results supported the clinical use of a standardized definition of centralization to facilitate patient classification and management and interpretation of outcomes. PMID- 18383646 TI - Injury patterns in elite preprofessional ballet dancers and the utility of screening programs to identify risk characteristics. AB - STUDY DESIGN: Retrospective descriptive cohort study. OBJECTIVES: To describe the distribution and rate of injuries in elite adolescent ballet dancers, and to examine the utility of screening data to distinguish between injured and noninjured dancers. BACKGROUND: Adolescent dancers account for most ballet injuries. Limited information exists, however, regarding the distribution of, rate of, and risk factors for, adolescent dance injuries. METHODS AND MEASURES: Two hundred four dancers (age, 9-20 years) were screened over 5 years. Screening data were collected at the beginning and injury data were collected at the end of each training year. Descriptive statistics were used to characterize distribution and rate of injuries. Inference statistics were used to examine differences between injured and noninjured dancers. RESULTS: Fifty-three percent of injuries occurred in the foot/ankle, 21.6% in the hip, 16.1% in the knee, and 9.4% in the back. Thirty-two to fifty-one percent of the dancers were injured each year, and, over the 5 years, there were 1.09 injuries per 1000 athletic exposures, and 0.77 injuries per 1000 hours of dance. Significant differences between injured and noninjured dancers were limited to current disability scores (P = .007), history of low back pain (P = .017), right foot pronation (P = .005), insufficient right ankle plantar flexion (P = .037), and lower extremity strength (P = .045). CONCLUSION: Distribution of injuries was similar to that of other studies. Injury rates were lower than most reported rates, except when expressed per 1000 hours of dance. Few differences were found between injured and noninjured dancers. These findings should be considered when designing and implementing screening programs. PMID- 18383647 TI - Differences in lower extremity anatomical and postural characteristics in males and females between maturation groups. AB - STUDY DESIGN: Descriptive, cross-sectional. OBJECTIVES: We compared lower extremity anatomical characteristics in males and females between different maturation groups. BACKGROUND: Sex differences have been observed in lower extremity anatomical characteristics. While the reasons contributing to these sex differences in adults are unknown, there is evidence that anatomy and posture change considerably during growth and development. METHODS AND MEASURES: One hundred seventy-three young athletes (age range, 9-18 years) were assessed for stage of maturation and placed into 1 of 3 groups, according to Tanners stages 1 and 2 (MatGrp1), 3 and 4 (MatGrp2), and 5 (MatGrp3). Participants were measured for pelvic angle, hip anteversion, quadriceps angle, tibiofemoral angle, femur length, tibial length, genu recurvatum, tibial torsion, navicular drop, general joint laxity, and anterior knee laxity. Data were compared by sex and maturation group. RESULTS: When comparing maturation groups, limb length, pelvic angle, and tibial torsion increased with maturation, and anterior knee laxity, genu recurvatum, tibiofemoral angle, and foot pronation decreased with maturation. Females had greater general joint laxity, hip anteversion, and tibiofemoral angles, and shorter femur and tibial lengths than males, regardless of maturation group. Maturational changes in knee laxity and quadriceps angles were sex dependent. CONCLUSIONS: We observed a general change of posture with maturation that began with greater knee valgus, knee recurvatum, and foot pronation in MatGrp1, then moved toward a relative straightening and external rotation of the knee, and supination of the foot in later maturation groups. While the majority of the measures changed similarly in males and females across maturation groups, decreases in quadriceps angles and anterior knee laxity were greater in males compared to females, and females were observed to have a more inwardly rotated hip and valgus knee posture, compared to males, particularly in later maturation groups. PMID- 18383648 TI - Liver damage with the amoxicillin-clavulanate combination. AB - Liver damage associated with the amoxicillin-clavulanate combination is more frequent in patients over the age of 50 and during long-term treatment. It is mainly due to the clavulanic acid component of the drug. It is better to reserve this combination for infections due to bacteria that are resistant to amoxicillin. PMID- 18383649 TI - Cholinesterase inhibitors: numerous and serious interactions: serious but preventable. PMID- 18383650 TI - Possible factors related to functional ankle instability. AB - STUDY DESIGN: Case control study. OBJECTIVES: To classify individuals with functional ankle instability (FAI) into deficit and non-deficit categories based on the evaluation of the most common factors that have been proposed to be related to FAI. BACKGROUND: Recent studies have suggested that FAI may be secondary to a combination of factors including ankle proprioceptive deficit, muscular weakness, impaired balance, delayed neuromuscular reaction time, and joint laxity. However, only a few authors have investigated the prevalence and association among these factors in a single group of individuals. METHODS AND MEASURES: The above 5 factors were tested bilaterally in 21 individuals with FAI and in 16 healthy control subjects. Data were analyzed for (1) within- and between-group comparison, (2) classification of subjects with FAI into deficit and no-deficit categories, and (3) magnitude of association between factors in the subjects with FAI using Pearson bivariate correlation. RESULTS: Balance control and evertors' strength were significantly less on the affected side in comparison to the unaffected side in subjects with FAI. The evertors' strength was also significantly different between the side difference of the FAI group and the side difference of the control group. Passive ankle stiffness was significantly correlated to balance control, ankle proprioception, and evertor peak torque. Individuals with FAI demonstrated a large variation in the deficit categories ranging from multiple deficits to no noticeable deficits. CONCLUSION: Mechanical alterations in the ankle joint may influence several aspects of the ankle's functional ability. Alterations in the afferent processes, represented in this study by ankle proprioception, may effect the evertors' strength or vice versa. More importantly, individuals with FAI might exhibit high variability in ankle deficits. PMID- 18383651 TI - Antidepressants: suicide among young adults: more frequent than with placebo. PMID- 18383652 TI - Psychoanalysis, history, and my own private Germany. PMID- 18383653 TI - Slipped capital femoral epiphysis in a patient referred to physical therapy for knee pain. PMID- 18383654 TI - Mammographic screening for breast cancer: few new data. AB - (1) A new comparative randomised trial of mammographic screening for breast cancer, involving about 160 000 women aged from 39 to 49, followed for more than 10 years, showed no significant reduction in breast cancer mortality. After seven screening mammograms, 23% of the women had at least one false-positive result. (2) The results of the Swedish Two-County trial were challenged after re examination of cancer registries. (3) One study showed that nearly 25% of women in whom breast cancer was detected with mammography in the Malmo-1 study would otherwise have remained asymptomatic for 25 years. (4) An update of the Cochrane meta-analysis confirmed previous conclusions: no tangible benefits were found in the 3 most reliable trials. When 5 less reliable trials were also taken into account, screening was found to reduce breast cancer mortality after 13 years of follow-up. (5) In France, there are no data on the impact of screening on the frequency of surgery or chemotherapy, the detection of harmless tumours or carcinoma in situ, or the risk of over-diagnosis. (6) In practice, healthcare professionals must inform their patients of the uncertainties concerning mammographic screening. PMID- 18383655 TI - The fire next time: the conversion of the Huli Apocalypse. PMID- 18383656 TI - Seasonal allergic rhinitis: limited effectiveness of treatments. AB - (1) Seasonal allergic rhinitis, otherwise known as hayfever, is a harmless condition, although it can cause major discomfort and interfere with activities of daily living. We conducted a review of the literature, based on our in-house methodology, to determine the risk-benefits of treatments used in this setting. (2) Placebo-controlled trials show that sodium cromoglicate relieves symptoms, especially if it is used before symptoms appear. Adverse effects are rare with sodium cromoglicate nasal solutions and eye drops. (3) Nasal steroids have well documented efficacy. Beclometasone is the best choice. Adverse effects include epistaxis, nasal irritation and, occasionally, systemic disorders. (4) Oral antihistamines are less effective than nasal steroids. They also provoke adverse effects, especially drowsiness. Nasal azelastine seems to have a similar efficacy as oral antihistamines. (5) The adverse effects of systemic steroids must not be overlooked, especially with long-term use. Oral administration is an alternative for severe symptoms that do not respond to other treatments, although this is rarely the case. Long-acting intramuscular steroids carry an increased risk of adverse effects. (6) Despite evaluation in several randomised controlled trials, there is no firm evidence that homeopathic preparations have any specific efficacy in allergic rhinitis. (7) Vasoconstrictors, ipratropium and montelukast, have negative risk-benefit balances in hay fever. (8) When a single allergen is responsible (grasses, ragweed, birch), clinical trials suggest that specific desensitisation can provide a modest improvement. However, this treatment carries a risk of local adverse effects, as well as a risk of rare but severe anaphylactic reactions, especially in patients who also have unstable severe asthma. (9) Sublingual desensitisation seems to be even less effective than subcutaneous desensitisation in adults. Follow-up is too short to know whether there is a risk of severe anaphylactic reactions. The results of paediatric studies are even less convincing. (10) In practice, when drug therapy is needed to relieve symptoms of seasonal allergic rhinitis, sodium cromoglicate is the first-line treatment. If a nasal steroid solution is chosen, it should be used for the shortest possible period. PMID- 18383657 TI - Oral mucositis due to cancer treatments. Orodental hygiene and ice cubes. AB - (1) Oral mucositis is a frequent adverse effect of cancer chemotherapy and radiotherapy. Fluorouracil, radiotherapy and conditioning regimens for haematopoietic stem cell grafting often cause severe oral mucositis, preventing patients from drinking and eating normally. (2) A randomised trial suggests that this complication can be attenuated by timely orodental care such as extraction of damaged teeth, treatment of tooth decay, and care of trauma due to dentures. (3) In 3 randomised controlled trials involving about 200 patients at high risk of severe oral mucositis, sucking ice during chemotherapy reduced the incidence of severe oral mucositis, from 14-74% to 4-21%. (4) Other treatments are no more effective and carry a risk of known or poorly documented adverse effects. (5) Analgesics (especially morphine) should be used to treat intense pain. Local anaesthetics have not been tested in patients with damaged oral mucosa, but they can cause a burning sensation and carry a risk of swallowing disorders due to anaesthesia of the oropharyngeal junction. (6) In practice, prevention of oral mucositis due to cancer chemotherapy or radiotherapy is based on orodental care and ice rather than drugs. PMID- 18383658 TI - Lymphatic filariasis. AB - (1) Lymphatic filariasis is a set of parasitic diseases that are endemic in tropical and subtropical regions and can be disabling in the long term. (2) The standard antiparasitic drug for adults is oral diethylcarbamazine. Ivermectin is an alternative, especially for patients with intercurrent onchocercosis or loasis. PMID- 18383659 TI - State and certifying boards/associations: CE and competency requirements. PMID- 18383660 TI - Point/Counterpoint. Cone beam x-ray CT will be superior to digital x-ray tomosynthesis in imaging the breast and delineating cancer. PMID- 18383661 TI - Shear strain imaging using shear deformations. AB - In this article we investigate the generation of shear strain elastograms induced using a lateral shear deformation. Ultrasound simulation and experimental results demonstrate that the shear strain elastograms obtained under shear deformation exhibit significant differences between bound and unbound inclusions in phantoms, when compared to shear strain images induced upon an axial compression. A theoretical model that estimates the decorrelation between pre- and postdeformation radio frequency signals, as a function of extent of shear deformation, is also developed. Signal-to-noise ratios of shear strain elastograms obtained at different shear angles are investigated theoretically and verified using ultrasound simulations on a uniformly elastic phantom. For the simulation and experiment, a two-dimensinal block-matching-based algorithm is used to estimate the axial and lateral displacement. Shear strains are obtained from the displacement vectors using a least-squares strain estimator. Our results indicate that the signal-to-noise ratio (SNR) of shear strain images increases to reach a maximum and saturates, and then decreases with increasing shear angle. Using typical system parameters, the maximum achievable SNR for shear strain elastography is around 8 (18 dB), which is comparable to conventional axial strain elastography induced by axial compression. Shear strain elastograms obtained experimentally using single inclusion tissue-mimicking phantoms with both bound and unbound inclusions (mimicking cancerous masses and benign fibroadenomas, respectively) demonstrate the characteristic differences in the depiction of these inclusions on the shear strain elastograms. PMID- 18383662 TI - Spin saturation artifact correction using slice-to-volume registration motion estimates for fMRI time series. AB - Evaluation of functional magnetic resonance imaging (fMRI) as a reliable clinical imaging tool requires accurate assessment and correction of head motion artifacts. As the correction of bulk head motion is vital, the loss of signal strength from the confounding effect of head motion on spin magnetization may be an additional factor in activation analysis error. This study focuses on the evaluation and correction of the spin saturation artifact that occurs when parts of adjacent slices are selected due to changing head positions in single-shot multislice acquisitions. As a consequence of head movement, the acquired slices constituting a fMRI volume are no longer parallel to each other and the spin magnetization in fMRI voxels becomes dependent on head motion history. Motion corrections applying the same rigid motion estimates to all the slices in a volume may not be a reasonable approximation in cases where the magnitude of head motion exceeds a subvoxel range. For realistic ranges of motion in fMRI, an accurate estimate of rigid motion parameters for each echo planar imaging (EPI) slice is essential to correctly register voxel intensities. Previously we have implemented the map-slice-to-volume (MSV) motion correction method that maps each slice in a time series onto a reference anatomical volume, which proved to be effective in improving activation detection. To correctly evaluate the motion dependence of spin magnetization, each voxel is tracked with movement history that is available from MSV motion estimates. Relatively low in resolution, EPI voxels are composed of varying mixtures of white matter (WM), gray matter (GM), and cerebrospinal fluid (CSF) and variations in the tissue composition give rise to voxel intensities that are functions of tissue T1 properties. We have developed a weighted-average spin saturation (WASS) correction method that can handle full rigid motion and account for the melange of different brain tissue isochromats at each EPI voxel location. We evaluated the effect of spin saturation artifacts and the performance of the WASS correction using simulated fMRI time series synthesized with known true activation, motion, and the associated spin saturation artifact. Two different ranges of head rotations, [ 5,5] and [-2,2] deg, were introduced and the effect of the spin saturation artifact was quantified to show 18% and 13% reduction in activation detection rate, respectively. Following the MSV motion and WASS correction, results indicate that WASS correction can improve activation detection by 17% relative to MSV only correction. PMID- 18383663 TI - Operating characteristics predicted by models for diagnostic tasks involving lesion localization. AB - In 1996 Swensson published an observer model that predicted receiver operating characteristic (ROC), localization ROC (LROC), free-response ROC (FROC) and alternative FROC (AFROC) curves, thereby achieving "unification" of different observer performance paradigms. More recently a model termed initial detection and candidate analysis (IDCA) has been proposed for fitting computer aided detection (CAD) generated FROC data, and recently a search model for human observer FROC data has been proposed. The purpose of this study was to derive IDCA and the search model based expressions for operating characteristics, and to compare the predictions to the Swensson model. For three out of four mammography CAD data sets all models yielded good fits in the high-confidence region, i.e., near the lower end of the plots. The search model and IDCA tended to better fit the data in the low-confidence region, i.e., near the upper end of the plots, particularly for FROC curves for which the Swensson model predictions departed markedly from the data. For one data set none of the models yielded satisfactory fits. A unique characteristic of search model and IDCA predicted operating characteristics is that the operating point is not allowed to move continuously to the lowest confidence limit of the corresponding Swensson model curves. This prediction is actually observed in the CAD raw data and it is the primary reason for the poor FROC fits of the Swensson model in the low-confidence region. PMID- 18383665 TI - Physical characteristics of GE Senographe Essential and DS digital mammography detectors. AB - The purpose of this study was to investigate physical characteristics of two full field digital mammography (FFDM) systems (GE Senographe Essential and DS). Both are indirect conversion (x ray to light) alpha-Si flat panels coupled with a CsI(Tl) scintillator. The examined systems have the same pixel size (100 microm) but a different field of view: a conventional size 23 x 19.2 cm2 and a large field 24 X 30.7 cm2, specifically designed to image large breasts. In the GE Senographe Essential model relevant improvements in flat panel design were implemented and new deposition tools for metal, alpha-Si, and CsI(Tl) were introduced by GE. These changes in detector design are expected to be beneficial for advanced applications such as breast tomosynthesis. The presampling modulation transfer function (MTF), normalized noise power spectrum (NNPS), and detective quantum efficiency (DQE) were measured for a wide range of exposure (25 240 microGy) with a RQA-M2 technique (28 kVp with a Mo/Mo target/filter combination and 2 mm of additional aluminum filtration). At 1, 2, and at 4 lp/mm MTF is equal to 0.9, 0.76, and 0.46 for the conventional field detector and to 0.85, 0.59, and 0.24 for the large field detector. The latter detector exhibits an improved NNPS due to a lower electronic noise and a better DQE that reaches 60%. In addition a contrast-detail analysis was performed with CDMAM 3.4 phantom and CDCOM software: GE Senographe DS showed statistically significant poorer detection ability in comparison with the GE Senographe Essential. These results could have been expected, at least qualitatively, considering the relative DQE of the two systems. PMID- 18383664 TI - Comparison of diffuse optical tomography of human breast with whole-body and breast-only positron emission tomography. AB - We acquire and compare three-dimensional tomographic breast images of three females with suspicious masses using diffuse optical tomography (DOT) and positron emission tomography (PET). Co-registration of DOT and PET images was facilitated by a mutual information maximization algorithm. We also compared DOT and whole-body PET images of 14 patients with breast abnormalities. Positive correlations were found between total hemoglobin concentration and tissue scattering measured by DOT, and fluorodeoxyglucose (18F-FDG) uptake. In light of these observations, we suggest potential benefits of combining both PET and DOT for characterization of breast lesions. PMID- 18383666 TI - Testing the GlaaS algorithm for dose measurements on low- and high-energy photon beams using an amorphous silicon portal imager. AB - The GLAaS algorithm for pretreatment intensity modulation radiation therapy absolute dose verification based on the use of amorphous silicon detectors, as described in Nicolini et al. [G. Nicolini, A. Fogliata, E. Vanetti, A. Clivio, and L. Cozzi, Med. Phys. 33, 2839-2851 (2006)], was tested under a variety of experimental conditions to investigate its robustness, the possibility of using it in different clinics and its performance. GLAaS was therefore tested on a low energy Varian Clinac (6 MV) equipped with an amorphous silicon Portal Vision PV aS500 with electronic readout IAS2 and on a high-energy Clinac (6 and 15 MV) equipped with a PV-aS1000 and IAS3 electronics. Tests were performed for three calibration conditions: A: adding buildup on the top of the cassette such that SDD-SSD = d(max) and comparing measurements with corresponding doses computed at d(max), B: without adding any buildup on the top of the cassette and considering only the intrinsic water-equivalent thickness of the electronic portal imaging devices device (0.8 cm), and C: without adding any buildup on the top of the cassette but comparing measurements against doses computed at d(max). This procedure is similar to that usually applied when in vivo dosimetry is performed with solid state diodes without sufficient buildup material. Quantitatively, the gamma index (gamma), as described by Low et al. [D. A. Low, W. B. Harms, S. Mutic, and J. A. Purdy, Med. Phys. 25, 656-660 (1998)], was assessed. The gamma index was computed for a distance to agreement (DTA) of 3 mm. The dose difference deltaD was considered as 2%, 3%, and 4%. As a measure of the quality of results, the fraction of field area with gamma larger than 1 (%FA) was scored. Results over a set of 50 test samples (including fields from head and neck, breast, prostate, anal canal, and brain cases) and from the long-term routine usage, demonstrated the robustness and stability of GLAaS. In general, the mean values of %FA remain below 3% for deltaD equal or larger than 3%, while they are slightly larger for deltaD = 2% with %FA in the range from 3% to 8%. Since its introduction in routine practice, 1453 fields have been verified with GLAaS at the authors' institute (6 MV beam). Using a DTA of 3 mm and a deltaD of 4% the authors obtained %FA = 0.9 +/- 1.1 for the entire data set while, stratifying according to the dose calculation algorithm, they observed: %FA = 0.7 +/- 0.9 for fields computed with the analytical anisotropic algorithm and %FA = 2.4 +/- 1.3 for pencil-beam based fields with a statistically significant difference between the two groups. If data are stratified according to field splitting, they observed %FA = 0.8 +/- 1.0 for split fields and 1.0 +/- 1.2 for nonsplit fields without any significant difference. PMID- 18383667 TI - A prototype quantitative film scanner for radiochromic film dosimetry. AB - We have developed a high resolution, quantitative, two-dimensional optical film scanner for use with a commercial high sensitivity radiochromic film (RCF) for measuring single fraction external-beam radiotherapy dose distributions. The film scanner was designed to eliminate artifacts commonly observed in RCF dosimetry. The scanner employed a stationary light source and detector with a moving antireflective glass film platen attached to a high precision computerized X-Y translation stage. An ultrabright red light emitting diode (LED) with a peak output at 633 nm and full width at half maximum (FWHM) of 16 nm was selected as the scanner light source to match the RCF absorption peak. A dual detector system was created using two silicon photodiode detectors to simultaneously measure incident and transmitted light. The LED light output was focused to a submillimeter (FWHM 0.67 mm) spot size, which was determined from a scanning knife-edge technique for measuring Gaussian optical beams. Data acquisition was performed with a 16-bit A/D card in conjunction with commercial software. The linearity of the measured densities on the scanner was tested using a calibrated neutral-density step filter. Sensitometric curves and three IMRT field scans were acquired with a spatial resolution of 1 mm for both radiographic film and RCF. The results were compared with measurements taken with a commercial diode array under identical delivery conditions. The RCF was rotated by 90 deg and rescanned to study orientation effects. Comparison between the RCF and the diode array measurements using percent dose difference and distance-to-agreement criteria produced average passing rates of 99.0% using 3%/3 mm criteria and 96.7% using 2%/2 mm criteria. The same comparison between the radiographic film and diode array measurements resulted in average passing rates 96.6% and 91.6% for the above two criteria, respectively. No measurable light-scatter or interference scanner artifacts were observed. The RCF rotated by 90 deg showed no measurable orientation effect. A scan of a 15 x 15 cm2 area with 1 mm resolution required 22 min to acquire. The LED densitometer provides an accurate film dosimetry system with 1 mm or better resolution. The scanner eliminates the orientation dependence of RCF dosimetry that was previously reported with commercial flatbed scanners. PMID- 18383668 TI - A dose comparison study between XVI and OBI CBCT systems. AB - The purpose of this study is to establish a comprehensive set of dose measurements data obtained from the X-ray Volumetric Imager (XVI, Elekta Oncology Systems) and the On-Board Imager (OBI, Varian Medical Systems) cone-beam CT (CBCT) systems. To this end, two uniform-density cylindrical acrylic phantoms with diameters of 18 cm (head phantom) and 30 cm (body phantom) were used for all measurements. Both phantoms included ion chamber placement holes in the center and at periphery (2 cm below surface). For the XVI unit, the four standard manufacturer-supplied protocols were measured. For the OBI unit, the full bow tie and half bow tie (and no bow tie) filters were used in combination with the two scanning modes; namely, full-fan and half-fan. The total milliampere x seconds (mA s) setting was also varied for each protocol to establish the linear relationship between the dose deposited and the mA s used (with all other factors being held constant). Half-value layers in aluminum (Al) were also measured for beam characteristic determination. For the XVI unit, the average dose ranged from 0.1 to 3.5 cGy with the highest dose measured using the "prostate" protocol with the body phantom. For the OBI unit, the average dose ranged from 1.1 to 8.3 cGy with the highest dose measured using the full-fan protocol with the head phantom. The measured doses were highly linear as a function of mA s, for both units, where the measurement points followed a linear relationship very closely with R2 > 0.99 for all cases. Half-value layers were between 4.6- and 7.0-mm-Al for the two CBCT units where XVI generally had more penetrating beams at the similar kVp settings. In conclusion, a comprehensive series of dose measurements were performed on the XVI and the OBI CBCT units. In the process, many of the important similarities and differences between the two systems were observed and summarized in this work. PMID- 18383669 TI - An ellipse-fitting based method for efficient registration of breast masses on two mammographic views. AB - When reading mammograms, radiologists routinely search for and compare suspicious breast lesions identified on two corresponding craniocaudal (CC) and mediolateral oblique (MLO) views. Automatically identifying and matching the same true positive breast lesions depicted on two views is an important step for developing successful multiview based computer-aided detection (CAD) schemes. The authors developed a method to automatically register breast areas and detect matching strips of interest used to identify the matched mass regions depicted on CC and MLO views. The method uses an ellipse based model to fit the breast boundary contour (skin line) and set a local Cartesian coordinate system for each view. One intersection point between the major/minor axis and the fitted ellipse perimeter passed through breast boundary is selected as the origin and the majoraxis and the minoraxis of the ellipse are used as the two axis of the Cartesian coordinate system. When a mass is identified on one view, the scheme computes its position in the local coordinate system. Then, the distance is mapped onto the local coordinate of the other view. At the end of the mapped distance a registered centerline of the matching strip is created. The authors established an image database that includes 200 test examinations each depicting one verified mass visible on the two views. They tested whether the registered centerline identified on another view can be used to locate the matched mass region. The experiments show that the average distance between the mass region centers and the registered centerlines was +/- 8.3 mm and in 91% of testing cases the registered centerline actually passes through the matched mass regions. A matching strip width of 47 mm was required to achieve 100% sensitivity for the test database. The results demonstrate the feasibility of the proposed method to automatically identify masses depicted on CC and MLO views, which may improve future development of multiview based CAD schemes. PMID- 18383670 TI - Design and evaluation of a HDR skin applicator with flattening filter. AB - The purposes of this study are: (i) to design field flattening filters for the Leipzig applicators of 2 and 3 cm of inner diameter with the source traveling parallel to the applicator contact surface, which are accessories of the microSelectron-HDR afterloader (Nucletron, Veenendaal, The Netherlands). These filters, made of tungsten, aim to flatten the heterogeneous dose distribution obtained with the Leipzig applicators. (ii) To estimate the dose rate distributions for these Leipzig+filter applicators by means of the Monte Carlo (MC) method. (iii) To experimentally verify these distributions for prototypes of these new applicators, and (iv) to obtain the correspondence factors to measure the output of the applicators by the user using an insert into a well chamber. The MC GEANT4 code has been used to design the filters and to obtain the dose rate distributions in liquid water for the two Leipzig+filter applicators. In order to validate this specific application and to guarantee that realistic source-applicator geometry has been considered, an experimental verification procedure was implemented in this study, in accordance with the updated recommendations of the American Association of Physicists in Medicine Task Group No. 43 U1 Report. Thermoluminescent dosimeters, radiochromic film, and a pin point ionization chamber in a plastic [polymethylmethacrylate (PMMA)] phantom were used to verify the MC results for the two applicators of a microSelectron HDR afterloader with the mHDR-v2 source. To verify the output of the Leipzig +filter applicators, correspondence factors were deduced for the well chambers HDR100-plus (Standard Imaging, Inc., Middleton, WI) and TM33004 (PTW, Freiburg, Germany) using a specific insert for both applicators. The doses measured in the PMMA phantom agree within experimental uncertainties with the dose obtained by the MC calculations. Percentage depth dose and off-axis profiles were obtained normalized at a depth of 3 mm along the central applicator axis in a cylindrical 20 x 20 cm water phantom. A table of output factors, normalized to 1 U of source air kerma strength at this depth, is presented. Correspondence factors were obtained for the two well chambers considered. The matrix data obtained in the MC simulation with a grid separation of 0.5 mm has been used to build a data set in a convenient format to model these distributions for routine use with a brachytherapy treatment planning system. PMID- 18383672 TI - Calibration of the Gamma Knife using a new phantom following the AAPM TG51 and TG21 protocols. AB - PURPOSE: To compare calibration of the Leksell Gamma Knife according to the American Association of Physicists in Medicine Task Groups 21 and 51 protocols. A new phantom was fabricated for this purpose. Its design, physical properties, and composition are described. MATERIALS AND METHODS: The Gamma Knife TG-51 calibration phantom is designed to be filled with water and support an ionization chamber positioned at its center. The phantom is thimble-shaped, with a 2 mm plastic wall to contain water. The phantom and chamber assembly was mounted in a Leksell stereotactic frame. The location of the chamber's sensitive volume was determined using computed tomography. The chamber-phantom assembly was attached to the 18 mm helmet in the Gamma Knife by the stereotactic frame. The phantom's geometry allowed radiation beams from each of the 201 Gamma Knife cobalt-60 sources to converge after an 8 cm path to the ionization chamber's sensitive volume. This is similar to the arrangement by which one calibrates the Gamma Knife using the manufacturer-supplied polystyrene phantom. RESULTS: The phantom was attached to the Gamma Knife so that the ionization chamber was reproducibly positioned at the convergence of the radiation beams. Because of the phantom's design, the phantom could be affixed to either trunnions or the automatic patient positioning system, once mounted in the Leksell stereotectic frame. Comparisons using different phantoms and protocols resulted in the following calibration ratios for TG-21 in the polystyrene sphere phantom, TG-21 in the water phantom, and TG-51 in the water phantom, respectively: 1.000, 1.008, 0.986, when corrected for transmission through the plastic water reservoir wall and using the same ionization chamber. Transmission measurements using a 1 cm thickness of the same material in the Co-60 beam determined that the phantom's 2 mm plastic wall resulted in a reduction in the measured the output by 0.5%. CONCLUSIONS: Calibration of the Gamma Knife can be performed in liquid water using the AAPM TG 51 protocol and this new phantom, thereby eliminating uncertainties with respect to the composition of the manufacturer's phantom. Perturbation of calibration measurements by nonwater materials was characterized and could be corrected. Calibration values for the Gamma Knife that were obtained using the three methods for our phantoms agree to within 1.4%. TG21 and TG51 calibration of the Gamma Knife using the water phantom agreed to within 2.2%. PMID- 18383671 TI - Total scatter factors of small beams: a multidetector and Monte Carlo study. AB - The scope of this study was to estimate total scatter factors (S(c,p)) of the three smallest collimators of the Cyberknife radiosurgery system (5-10 mm in diameter), combining experimental measurements and Monte Carlo simulation. Two microchambers, a diode, and a diamond detector were used to collect experimental data. The treatment head and the detectors were simulated by means of a Monte Carlo code in order to calculate correction factors for the detectors and to estimate total scatter factors by means of a consistency check between measurement and simulation. Results for the three collimators were: S(c,p) (5 mm) = 0.677 +/- 0.004, S(c,p) (7.5 mm) = 0.820 +/- 0.008, S(c,p) (10 mm) = 0.871 +/- 0.008, all relative to the 60 mm collimator at 80 cm source-to-detector distance. The method also allows the full width at half maximum of the electron beam to be estimated; estimations made with different collimators and different detectors were in excellent agreement and gave a value of 2.1 mm. Correction factors to be applied to the detectors for the measurement of S(c,p) were consistent with a prevalence of volume effect for the microchambers and the diamond and a prevalence of scattering from high-Z material for the diode detector. The proposed method is more sensitive to small variations of the electron beam diameter with respect to the conventional method used to commission Monte Carlo codes, i.e., by comparison with measured percentage depth doses (PDD) and beam profiles. This is especially important for small fields (less than 10 mm diameter), for which measurements of PDD and profiles are strongly affected by the type of detector used. Moreover, this method should allow S(c,p) of Cyberknife systems different from the unit under investigation to be estimated without the need for further Monte Carlo calculation, provided that one of the microchambers or the diode detector of the type used in this study are employed. The results for the diamond are applicable only to the specific detector that was investigated due to excessive variability in manufacturing. PMID- 18383673 TI - Image deconvolution in digital autoradiography: a preliminary study. AB - Digital autoradiography (DAR) is a powerful method to determine quantitatively the "small-scale" (i.e., submillimeter) distribution of a radiotracer within a tissue section. However, the limited spatial resolution of the DAR image, due to blurring by the point spread function (PSF), can result in a poor correlation with tissue histology and immunohistochemistry. The authors attempt to overcome this limitation by recovering the radiotracer distribution by image deconvolution using the Richardson-Lucy algorithm and a measured PSF obtained from a small radioactive source on hydrophobic microscope slide. Simulation studies have shown that the deconvolution algorithm reliably recovers the pixel values corresponding to the radioactivity distributions. As an example, the proposed image restoration approach has been tested with DAR images of different radiolabeled markers on tumor sections obtained from clinical and preclinical animal model studies. Digital autoradiograms following deconvolution show improved sharpness and contrast relative to the unprocessed autoradiograms. PMID- 18383674 TI - Comparison of mega-voltage cone-beam computed tomography prostate localization with online ultrasound and fiducial markers methods. AB - The online image-guided localization data from 696 ultrasound (US), 598 mega voltage cone-beam computed tomography (MV-CBCT), and 393 seed markers (SMs) couch alignments for patients undergoing intensity modulation radiotherapy of the prostate were analyzed. Daily US, MV-CBCT and SM images were acquired for 19, 17 and 12 patients, respectively, after each patient was immobilized in a vacuum cradle and setup to skin markers as the center of mass. The couch shifts applied in the lateral (left-right/LR), vertical (anterior-posterior/AP), and longitudinal (superior-inferior/SI) directions, along with the magnitude of the three-dimensional (3D) shift vector, were analyzed and compared for all three methods. The percentage of shifts larger than 5 mm in all directions was also compared. Clinical target volume-planning target volume (CTV-to-PTV) expansion margins were estimated based on the localization data with US, CB, and SM image guidance. Results show the US data have greater variability. Systematic and random shifts were -1.2 +/- 6.8 mm (LR), -2.8 +/- 5.1 mm (SI) and -1.0 +/- 5.9 mm (AP) for US, 1.0 +/- 3.9 mm (LR), -1.3 +/- 2.5 mm (SI) and -0.3 +/- 3.9 mm (AP) for CB, and -1.0 +/- 3.4 mm (LR), 0.0 +/- 3.4 mm (SI) and 0.5 +/- 4.1 mm (AP) for SM. The mean 3D shift distance was larger using US (8.8 +/- 6.2 mm) compared to CB and SM (5.3 +/- 3.4 mm and 5.2 +/- 3.7 mm, respectively). The percentage of US shifts larger than 5 mm were 34%, 31%, and 38% in the LR, SI, and AP directions, respectively, compared to 18%, 6%, and 16% for CB and 14%, 10%, and 20% for SM. MV-CBCT and SM localization data suggest a different distribution of prostate center-of-mass shifts with smaller variability, compared to US. The online MV CBCT and SM image-guidance data show that for treatments that do not include daily prostate localization, one can use a CTV-to-PTV margin that is 4 mm smaller than the one suggested by US data, hence allowing more rectum and bladder sparing and potentially improving the therapeutic ratio. PMID- 18383675 TI - Vascular oxygen content and the tissue oxygenation--a theoretical analysis. AB - Several methods exist for evaluating tumor oxygenation as hypoxia is an important prognostic factor for cancer patients. They use different measuring principles that highlight various aspects of oxygenation. The results could be empirically correlated, but it has been suspected that there could be discordances in some cases. This study describes an analysis of the relationship between vascular and tissue oxygenations. Theoretical simulation has been employed to characterize tissue oxygenations for a broad range of distributions of intervessel distances and vascular oxygenations. The results were evaluated with respect to the implications for practical measurements of tissue oxygenations. The findings showed that although the tissue oxygenation is deterministically related to vascular oxygenation, the relationship between them is not unequivocal. Variability also exists between the fractions of values below the sensitivity thresholds of various measurement methods which in turn could be reflected in the power of correlations between results from different methods or in the selection of patients for prognostic studies. The study has also identified potential difficulties that may be encountered at the quantitative evaluation of the results from oxygenation measurements. These could improve the understanding of oxygenation measurements and the interpretation of comparisons between results from various measurement methods. PMID- 18383676 TI - Accurate registration of random radiographic projections based on three spherical references for the purpose of few-view 3D reconstruction. AB - Precise registration of radiographic projection images acquired in almost arbitrary geometries for the purpose of three-dimensional (3D) reconstruction is beset with difficulties. We modify and enhance a registration method [R. Schulze, D. D. Bruellmann, F. Roeder, and B. d'Hoedt, Med. Phys. 31, 2849-2854 (2004)] based on coupling a minimum amount of three reference spheres in arbitrary positions to a rigid object under study for precise a posteriori pose estimation. Two consecutive optimization procedures (a, initial guess; b, iterative coordinate refinement) are applied to completely exploit the reference's shadow information for precise registration of the projections. The modification has been extensive, i.e., only the idea of using the sphere shadows to locate each sphere in three dimensions from each projection was retained whereas the approach to extract the shadow information has been changed completely and extended. The registration information is used for subsequent algebraic reconstruction of the 3D information inherent in the projections. We present a detailed mathematical theory of the registration process as well as simulated data investigating its performance in the presence of error. Simulation of the initial guess revealed a mean relative error in the critical depth coordinate ranging between 2.1% and 4.4%, and an evident error reduction by the subsequent iterative coordinate refinement. To prove the applicability of the method for real-world data, algebraic 3D reconstructions from few (< or = 9) projection radiographs of a human skull, a human mandible and a teeth-containing mandible segment are presented. The method facilitates extraction of 3D information from only few projections obtained from off-the-shelf radiographic projection units without the need for costly hardware. Technical requirements as well as radiation dose are low. PMID- 18383677 TI - Calculation of organ doses in x-ray examinations of premature babies. AB - Lung disease represents one of the most life-threatening conditions in prematurely born children. In the evaluation of the neonatal chest, the primary and most important diagnostic study is the chest radiograph. Since prematurely born children are very sensitive to radiation, those radiographs may lead to a significant radiation detriment. Knowledge of the radiation dose is therefore necessary to justify the exposures. To calculate doses in the entire body and in specific organs, computational models of the human anatomy are needed. Using medical imaging techniques, voxel phantoms have been developed to achieve a representation as close as possible to the anatomical properties. In this study two voxel phantoms, representing prematurely born babies, were created from computed tomography- and magnetic resonance images: Phantom 1 (1910 g) and Phantom 2 (590 g). The two voxel phantoms were used in Monte Carlo calculations (MCNPX) to assess organ doses. The results were compared with the commercially available software package PCXMC in which the available mathematical phantoms can be downsized toward the prematurely born baby. The simple phantom-scaling method used in PCXMC seems to be sufficient to calculate doses for organs within the radiation field. However, one should be careful in specifying the irradiation geometry. Doses in organs that are wholly or partially outside the primary radiation field depend critically on the irradiation conditions and the phantom model. PMID- 18383678 TI - Evaluation of dosimetric margins in prostate IMRT treatment plans. AB - This work introduces a new concept--the dosimetric margin distribution (DMD)--and uses it to explain the sensitivity of a group of prostate IMRT treatment plans to patient setup errors. Prior work simulated the effect of setup errors on 27 prostate IMRT treatment plans and found the plans could tolerate larger setup errors than predicted by the van Herk margin formula. The conjectured reason for this disagreement was a breakdown in van Herk's assumption that the planned dose distribution conforms perfectly to target structures. To resolve the disagreement, this work employed the same 27 plans to evaluate the actual margin distributions that exist between: (i) the clinical target volume (CTV) and planning target volume (PTV) and (ii) the CTV and PTV minimum dose isodose surface. These distributions were evaluated for both prostate and nodal targets. Distribution (ii) is the DMD. The dosimetric margin in a given direction determines the probability that the CTV will be underdosed due to setup errors in that direction. Averaging over 4 pi sr gives the overall probability of CTV coverage. Minimum doses for prostate and nodal PTVs were obtained from dose volume histograms. Corresponding isodose surfaces were created and converted to regions of interest (ROIs). CTV, PTV, and isodose ROIs were saved as mesh files and then imported into a computational geometry application which calculated distances between meshes (i.e., margins) in 614 discrete directions covering 4 pi sr in 10 deg increments. Measured prostate CTV-to-PTV margins were close to the nominal value of 0.5 cm specified in the treatment planning protocol. However, depending on direction, prostate dosimetric margins ranged from 0.5 to 3 cm, reflecting the imperfect conformance of the planned dose distribution to the prostate PTV. For the nodal CTV, the nominal CTV-to-PTV margin employed in treatment planning was again 0.5 cm. However, due to the planning protocol, the nodal PTV follows the surface of the nodal CTV in several places, ensuring that there is no room for rigid body motion of the nodal CTV inside the nodal PTV. Measured nodal CTV-to-PTV margins were therefore zero, while nodal dosimetric margins ranged from 0.2 to 2.8 cm. Prostate and nodal target coverage were found to be well correlated with the measured DMDs, thereby resolving the apparent disagreement with our prior results. The principal conclusion is that target coverage in the presence of setup errors should be evaluated using the DMD, rather than the CTV-to-PTV margin distribution. The DMD is a useful planning metric, which generalizes the ICRU conformity index. DMDs could vary with number of beams, beam arrangements, TPS, and treatment site. PMID- 18383679 TI - Respiratory-gated CT as a tool for the simulation of breathing artifacts in PET and PET/CT. AB - Respiratory motion in PET and PET/CT blurs the images and can cause attenuation related errors in quantitative parameters such as standard uptake values. In rare instances, this problem even causes localization errors and the disappearance of tumors that should be detectable. Attenuation errors are severe near the diaphragm and can be enhanced when the attenuation correction is based on a CT series acquired during a breath-hold. To quantify the errors and identify the parameters associated with them, the authors performed a simulated PET scan based on respiratory-gated CT studies of five lung cancer patients. Diaphragmatic motion ranged from 8 to 25 mm in the five patients. The CT series were converted to 511-keV attenuation maps which were forward-projected and exponentiated to form sinograms of PET attenuation factors at each phase of respiration. The CT images were also segmented to form a PET object, moving with the same motion as the CT series. In the moving PET object, spherical 20 mm mobile tumors were created in the vicinity of the dome of the liver and immobile 20 mm tumors in the midchest region. The moving PET objects were forward-projected and attenuated, then reconstructed in several ways: phase-matched PET and CT, gated PET with ungated CT, ungated PET with gated CT, and conventional PET. Spatial resolution and statistical noise were not modeled. In each case, tumor uptake recovery factor was defined by comparing the maximum reconstructed pixel value with the known correct value. Mobile 10 and 30 mm tumors were also simulated in the case of a patient with 11 mm of breathing motion. Phase-matched gated PET and CT gave essentially perfect PET reconstructions in the simulation. Gated PET with ungated CT gave tumors of the correct shape, but recovery was too large by an amount that depended on the extent of the motion, as much as 90% for mobile tumors and 60% for immobile tumors. Gated CT with ungated PET resulted in blurred tumors and caused recovery errors between -50% and +75%. Recovery in clinical scans would be 0%-20% lower than stated because spatial resolution was not included in the simulation. Mobile tumors near the dome of the liver were subject to the largest errors in either case. Conventional PET for 20 mm tumors was quantitative in cases of motion less than 15 mm because of canceling errors in blurring and attenuation, but the recovery factors were too low by as much as 30% in cases of motion greater than 15 mm. The 10 mm tumors were blurred by motion to a greater extent, causing a greater SUV underestimation than in the case of 20 mm tumors, and the 30 mm tumors were blurred less. Quantitative PET imaging near the diaphragm requires proper matching of attenuation information to the emission information. The problem of missed tumors near the diaphragm can be reduced by acquiring attenuation-correction information near end expiration. A simple PET/CT protocol requiring no gating equipment also addresses this problem. PMID- 18383680 TI - Cascaded systems analysis of noise reduction algorithms in dual-energy imaging. AB - An important aspect of dual-energy (DE) x-ray image decomposition is the incorporation of noise reduction techniques to mitigate the amplification of quantum noise. This article extends cascaded systems analysis of imaging performance to DE imaging systems incorporating linear noise reduction algorithms. A general analytical formulation of linear DE decomposition is derived, with weighted log subtraction and several previously reported noise reduction algorithms emerging as special cases. The DE image noise-power spectrum (NPS) and modulation transfer function (MTF) demonstrate that noise reduction algorithms impart significant, nontrivial effects on the spatial-frequency dependent transfer characteristics which do not cancel out of the noise equivalent quanta (NEQ). Theoretical predictions were validated in comparison to the measured NPS and MTF. The resulting NEQ was integrated with spatial-frequency dependent task functions to yield the detectability index, d', for evaluation of DE imaging performance using different decomposition algorithms. For a 3 mm lung nodule detection task, the detectability index varied from d' < 1 (i.e., nodule barely visible) in the absence of noise reduction to d' > 2.5 (i.e., nodule clearly visible) for "anti-correlated noise reduction" (ACNR) or "simple smoothing of the high-energy image" (SSH) algorithms applied to soft-tissue or bone-only decompositions, respectively. Optimal dose allocation (A*, the fraction of total dose delivered in the low-energy projection) was also found to depend on the choice of noise reduction technique. At fixed total dose, multi-function optimization suggested a significant increase in optimal dose allocation from A* = 0.32 for conventional log subtraction to A* = 0.79 for ACNR and SSH in soft tissue and bone-only decompositions, respectively. Cascaded systems analysis extended to the general formulation of DE image decomposition provided an objective means of investigating DE imaging performance across a broad range of acquisition and decomposition algorithms in a manner that accounts for the spatial-frequency-dependent imaging task. PMID- 18383681 TI - The grid-dose-spreading algorithm for dose distribution calculation in heavy charged particle radiotherapy. AB - A new variant of the pencil-beam (PB) algorithm for dose distribution calculation for radiotherapy with protons and heavier ions, the grid-dose spreading (GDS) algorithm, is proposed. The GDS algorithm is intrinsically faster than conventional PB algorithms due to approximations in convolution integral, where physical calculations are decoupled from simple grid-to-grid energy transfer. It was effortlessly implemented to a carbon-ion radiotherapy treatment planning system to enable realistic beam blurring in the field, which was absent with the broad-beam (BB) algorithm. For a typical prostate treatment, the slowing factor of the GDS algorithm relative to the BB algorithm was 1.4, which is a great improvement over the conventional PB algorithms with a typical slowing factor of several tens. The GDS algorithm is mathematically equivalent to the PB algorithm for horizontal and vertical coplanar beams commonly used in carbon-ion radiotherapy while dose deformation within the size of the pristine spread occurs for angled beams, which was within 3 mm for a single 150-MeV proton pencil beam of 30 degrees incidence, and needs to be assessed against the clinical requirements and tolerances in practical situations. PMID- 18383682 TI - In vivo assessment of magnesium status in human body using accelerator-based neutron activation measurement of hands: a pilot study. AB - Magnesium (Mg) is an element essential for many enzymatic reactions in the human body. Various human and animal studies suggest that changes in Mg status are linked to diseases such as cardiac arrhythmia, coronary heart disease, hypertension, premenstrual syndrome, and diabetes mellitus. Thus, knowledge of Mg levels in the human body is needed. A direct measurement of human blood serum, which contains only 0.3% of the total body Mg, is generally used to infer information about the status of Mg in the body. However, in many clinical situations, Mg stored in large levels, for example in bones, muscles, and soft tissues, needs to be monitored either to evaluate the efficacy of a treatment or to study the progression of diseases associated with the deficiency of total body Mg. This work presents a feasibility study of a noninvasive, in vivo neutron activation analysis (IVNAA) technique using the 26Mg (n, gamma) 27Mg reaction to measure Mg levels in human hands. The technique employs the McMaster University high beam current Tandetron accelerator hand irradiation facility and an array of eight NaI (T1) detectors arranged in a 4 pi geometry for delayed counting of the 0.844 and 1.014 MeV gamma rays emitted when 27Mg decays in the irradiated hand. Mg determination in humans using IVNAA of hands has been demonstrated to be feasible, with effective doses as low as one-quarter of those delivered in chest x rays. The overall experimental uncertainty in the measurements is estimated to be approximately 5% (1 sigma). The results are found to be in the range of the in vitro measurements reported for other cortical bones collected from different sites of the human skeleton, which confirms that this technique mainly provides a measure of the amount of Mg in hand bones. The average concentration of Mg determined in human hands is 10.96 +/- 1.25 (+/- 1 SD) mg Mg/g Ca. The coefficient of variation (11%) observed in this study is comparable with or lower than several studies using in vitro measurements reported in the literature and therefore allows for a quantitative intersubject comparison, even if to a limited extent. The features of the developed technique such as its simplicity, rapidity, accuracy, robustness, noninvasive nature, and very effective use of radiation doses, present the technique as a viable diagnostic tool available for trial in a clinical environment. PMID- 18383683 TI - A needle guidance system for biopsy and therapy using two-dimensional ultrasound. AB - Image-guided needle biopsies are currently used to provide a definitive diagnosis of breast cancer; however, difficulties in tumor targeting exist as the ultrasound (US) scan plane and biopsy needle must remain coplanar throughout the procedure to display the actual needle tip position. The additional time associated with aligning and maintaining this coplanar relationship results in increased patient discomfort. Biopsy procedural efficiency is further hindered since needle pathway interpretation is often difficult, especially for needle insertions at large depths that usually require multiple reinsertions. The authors developed a system that would increase the speed and accuracy of current breast biopsy procedures using readily available two-dimensional (2D) US technology. This system is composed of a passive articulated mechanical arm that attaches to a 2D US transducer. The arm is connected to a computer through custom electronics and software, which were developed as an interface for tracking the positioning of the mechanical components in real time. The arm couples to the biopsy needle and provides visual guidance for the physician performing the procedure in the form of a real-time projected needle pathway overlay on an US image of the breast. An agar test phantom, with stainless steel targets interspersed randomly throughout, was used to validate needle trajectory positioning accuracy. The biopsy needle was guided by both the software and hardware components to the targets. The phantom, with the needle inserted and device decoupled, was placed in an x-ray stereotactic mammography (SM) machine. The needle trajectory and bead target locations were determined in three dimensions from the SM images. Results indicated a mean needle trajectory accuracy error of 0.75 +/- 0.42 mm. This is adequate to sample lesions that are < 2 mm in diameter. Chicken tissue test phantoms were used to compare core needle biopsy procedure times between experienced radiologists and inexperienced resident radiologists using free-hand US and the needle guidance system. Cylindrical polyvinyl alcohol cryogel lesions, colored blue, were embedded in chicken tissue. Radiologists identified the lesions, visible as hypoechoic masses in the US images, and performed biopsy using a 14-gauge needle. Procedure times were compared based on experience and the technique performed. Using a pair-wise t test, lower biopsy procedure times were observed when using the guidance system versus the free-hand technique (t = 12.59, p < 0.001). The authors believe that with this improved biopsy guidance they will be able to reduce the "false negative" rate of biopsies, especially in the hands of less experienced physicians. PMID- 18383684 TI - Dual-energy imaging of the chest: optimization of image acquisition techniques for the 'bone-only' image. AB - Experiments were conducted to determine optimal acquisition techniques for bone image decompositions for a prototype dual-energy (DE) imaging system. Technique parameters included kVp pair (denoted [kVp(L)/kVp(H)]) and dose allocation (the proportion of dose in low- and high-energy projections), each optimized to provide maximum signal difference-to-noise ratio in DE images. Experiments involved a chest phantom representing an average patient size and containing simulated ribs and lung nodules. Low- and high-energy kVp were varied from 60-90 and 120-150 kVp, respectively. The optimal kVp pair was determined to be [60/130] kVp, with image quality showing a strong dependence on low-kVp selection. Optimal dose allocation was approximately 0.5-i.e., an equal dose imparted by the low- and high-energy projections. The results complement earlier studies of optimal DE soft-tissue image acquisition, with differences attributed to the specific imaging task. Together, the results help to guide the development and implementation of high-performance DE imaging systems, with applications including lung nodule detection and diagnosis, pneumothorax identification, and musculoskeletal imaging (e.g., discrimination of rib fractures from metastasis). PMID- 18383685 TI - Increasing the speed of DOSXYZnrc Monte Carlo simulations through the introduction of nonvoxelated geometries. AB - This article presents a method for increasing the speed of DOSXYZnrc Monte Carlo simulations through the introduction of nonvoxelated geometries defined in any coordinate system. Nonvoxelated geometries are used to isolate regions of uniform density and composition from the scoring grid. Particle transport within these geometric regions is not restricted by the boundary constraints of the scoring grid. This allows for larger particle steps, which in turn reduces the calculation time. A water tank phantom, water-lung interface phantom, cylindrical calibration phantom, and CT phantom were each used to test the application of the nonvoxelated approach. Each phantom was simulated using both the original DOSXYZnrc code and the new nonvoxelated code. The equivalence between the original and nonvoxelated simulations were quantified using a chi2 analysis. To within the statistical uncertainty, the voxelated and nonvoxelated simulations were found to give nearly identical results, regardless of boundary crossing algorithm. The speed increase was found to be a function of both voxel dimension and field size. Using nonvoxelated geometries and the EXACT boundary crossing algorithm, the speed increase was as high as 9.0, 5.1, 5.7, and 1.3 times faster for the water tank, water-lung interface, cylindrical calibration, and CT phantoms, respectively. If the PRESTA-I boundary crossing algorithm was used, the calculation speed increase was up to 6.0, 2.7, 3.3, and 1.2 times faster. These results clearly show that the nonvoxelated technique greatly increases simulation speed without any loss in dose accuracy. PMID- 18383686 TI - Anniversary paper: evaluation of medical imaging systems. AB - Medical imaging used to be primarily within the domain of radiology, but with the advent of virtual pathology slides and telemedicine, imaging technology is expanding in the healthcare enterprise. As new imaging technologies are developed, they must be evaluated to assess the impact and benefit on patient care. The authors review the hierarchical model of the efficacy of diagnostic imaging systems by Fryback and Thornbury [Med. Decis. Making 11, 88-94 (1991)] as a guiding principle for system evaluation. Evaluation of medical imaging systems encompasses everything from the hardware and software used to acquire, store, and transmit images to the presentation of images to the interpreting clinician. Evaluation of medical imaging systems can take many forms, from the purely technical (e.g., patient dose measurement) to the increasingly complex (e.g., determining whether a new imaging method saves lives and benefits society). Evaluation methodologies cover a broad range, from receiver operating characteristic (ROC) techniques that measure diagnostic accuracy to timing studies that measure image-interpretation workflow efficiency. The authors review briefly the history of the development of evaluation methodologies and review ROC methodology as well as other types of evaluation methods. They discuss unique challenges in system evaluation that face the imaging community today and opportunities for future advances. PMID- 18383687 TI - Prior image constrained compressed sensing (PICCS): a method to accurately reconstruct dynamic CT images from highly undersampled projection data sets. AB - When the number of projections does not satisfy the Shannon/Nyquist sampling requirement, streaking artifacts are inevitable in x-ray computed tomography (CT) images reconstructed using filtered backprojection algorithms. In this letter, the spatial-temporal correlations in dynamic CT imaging have been exploited to sparsify dynamic CT image sequences and the newly proposed compressed sensing (CS) reconstruction method is applied to reconstruct the target image sequences. A prior image reconstructed from the union of interleaved dynamical data sets is utilized to constrain the CS image reconstruction for the individual time frames. This method is referred to as prior image constrained compressed sensing (PICCS). In vivo experimental animal studies were conducted to validate the PICCS algorithm, and the results indicate that PICCS enables accurate reconstruction of dynamic CT images using about 20 view angles, which corresponds to an under sampling factor of 32. This undersampling factor implies a potential radiation dose reduction by a factor of 32 in myocardial CT perfusion imaging. PMID- 18383688 TI - Automatic registration between reference and on-board digital tomosynthesis images for positioning verification. AB - The authors developed a hybrid multiresolution rigid-body registration technique to automatically register reference digital tomosynthesis (DTS) images with on board DTS images to guide patient positioning in radiation therapy. This hybrid registration technique uses a faster but less accurate static method to achieve an initial registration, followed by a slower but more accurate adaptive method to fine tune the registration. A multiresolution scheme is employed in the registration to further improve the registration accuracy, robustness, and efficiency. Normalized mutual information is selected as the criterion for the similarity measure and the downhill simplex method is used as the search engine. This technique was tested using image data both from an anthropomorphic chest phantom and from eight head-and-neck cancer patients. The effects of the scan angle and the region-of-interest (ROI) size on the registration accuracy and robustness were investigated. The necessity of using the adaptive registration method in the hybrid technique was validated by comparing the results of the static method and the hybrid method. With a 44 degrees scan angle and a large ROI covering the entire DTS volume, the average of the registration capture ranges in single-axis simulations was between -31 and +34 deg for rotations and between -89 and +78 mm for translations in the phantom study, and between -38 and +38 deg for rotations and between -58 and +65 mm for translations in the patient study. Decreasing the DTS scan angle from 44 degrees to 22 degrees mainly degraded the registration accuracy and robustness for the out-of-plane rotations. Decreasing the ROI size from the entire DTS volume to the volume surrounding the spinal cord reduced the capture ranges to between -23 and +18 deg for rotations and between 33 and +43 mm for translations in the phantom study, and between -18 and +25 deg for rotations and between -35 and +39 mm for translations in the patient study. Results also showed that the hybrid registration technique had much larger capture ranges than the static method alone in registering the out-of-plane rotations. PMID- 18383689 TI - A pilot study exploring the possibility of establishing guidance levels in x-ray directed interventional procedures. AB - This article summarizes the dosimetric results of an International Atomic Energy Agency coordinated research program to investigate the feasibility of adopting guidance levels for invasive coronary artery procedures. The main study collected clinical data from hospitals located in five countries. A total of 2265 coronary angiograms (CA) and 1844 percutaneous coronary interventions (PCI) were analyzed. Substudies evaluated the dosimetric performance of 14 fluoroscopes, skin dose maps obtained using film, the quality of CA procedures, and the complexity of PCI procedures. Kerma-area product (PKA) guidance levels of 50 and 125 Gy cm2 are suggested for CA and PCI procedures. These levels should be adjusted for the complexity of the procedures performed in a given institution. PMID- 18383690 TI - Radiative transport in the delta-P1 approximation for semi-infinite turbid media. AB - We have developed an analytic solution for spatially resolved diffuse reflectance within the deltaP1 approximation to the radiative transport equation for a semi infinite homogeneous turbid medium. We evaluate the performance of this solution by comparing its predictions with those provided by Monte Carlo simulations and the standard diffusion approximation. We demonstrate that the delta-P1 approximation provides accurate estimates for spatially resolved diffuse reflectance in both low and high scattering media. We also develop a multi-stage nonlinear optimization algorithm in which the radiative transport estimates provided by the delta-P1 approximation are used to recover the optical absorption (microa), reduced scattering (micros'), and single-scattering asymmetry coefficients (g1) of liquid and solid phantoms from experimental measurements of spatially resolved diffuse reflectance. Specifically, the delta-P1 approximation can be used to recover microa, micros', and g1 with errors within +/- 22%, +/- 18%, and +/- 17%, respectively, for both intralipid-based and siloxane-based tissue phantoms. These phantoms span the optical property range 4 < (micros' /microa) < 117. Using these same measurements, application of the standard diffusion approximation resulted in the recovery of microa and micros' with errors o f +/- 29% and +/- 25%, respectively. Collectively, these results demonstrate that the delta-P1 approximation provides accurate radiative transport estimates that can be used to determine accurately the optical properties of biological tissues, particularly in spectral regions where tissue may display moderate/low ratios of reduced scattering to absorption (micros'/microa). PMID- 18383691 TI - Mixture of expert 3D massive-training ANNs for reduction of multiple types of false positives in CAD for detection of polyps in CT colonography. AB - One of the major challenges in computer-aided detection (CAD) of polyps in CT colonography (CTC) is the reduction of false-positive detections (FPs) without a concomitant reduction in sensitivity. A large number of FPs is likely to confound the radiologist's task of image interpretation, lower the radiologist's efficiency, and cause radiologists to lose their confidence in CAD as a useful tool. Major sources of FPs generated by CAD schemes include haustral folds, residual stool, rectal tubes, the ileocecal valve, and extra-colonic structures such as the small bowel and stomach. Our purpose in this study was to develop a method for the removal of various types of FPs in CAD of polyps while maintaining a high sensitivity. To achieve this, we developed a "mixture of expert" three dimensional (3D) massive-training artificial neural networks (MTANNs) consisting of four 3D MTANNs that were designed to differentiate between polyps and four categories of FPs: (1) rectal tubes, (2) stool with bubbles, (3) colonic walls with haustral folds, and (4) solid stool. Each expert 3D MTANN was trained with examples from a specific non-polyp category along with typical polyps. The four expert 3D MTANNs were combined with a mixing artificial neural network (ANN) such that different types of FPs could be removed. Our database consisted of 146 CTC datasets obtained from 73 patients whose colons were prepared by standard pre colonoscopy cleansing. Each patient was scanned in both supine and prone positions. Radiologists established the locations of polyps through the use of optical-colonoscopy reports. Fifteen patients had 28 polyps, 15 of which were 5-9 mm and 13 were 10-25 mm in size. The CTC cases were subjected to our previously reported CAD method consisting of centerline-based extraction of the colon, shape based detection of polyp candidates, and a Bayesian-ANN-based classification of polyps. The original CAD method yielded 96.4% (27/28) by-polyp sensitivity with an average of 3.1 (224/73) FPs per patient. The mixture of expert 3D MTANNs removed 63% (142/224) of the FPs without the loss of any true positive; thus, the FP rate of our CAD scheme was improved to 1.1 (82/73) FPs per patient while the original sensitivity was maintained. By use of the mixture of expert 3D MTANNs, the specificity of a CAD scheme for detection of polyps in CTC was substantially improved while a high sensitivity was maintained. PMID- 18383692 TI - Accurate dosimetry with GafChromic EBT film of a 6 MV photon beam in water: what level is achievable? AB - This paper focuses on the accuracy, in absolute dose measurements, with GafChromicTM EBT film achievable in water for a 6 MV photon beam up to a dose of 2.3 Gy. Motivation is to get an absolute dose detection system to measure up dose distributions in a (water) phantom, to check dose calculations. An Epson 1680 color (red green blue) transmission flatbed scanner has been used as film scanning system, where the response in the red color channel has been extracted and used for the analyses. The influence of the flatbed film scanner on the film based dose detection process was investigated. The scan procedure has been optimized; i.e. for instance a lateral correction curve was derived to correct the scan value, up to 10%, as a function of optical density and lateral position. Sensitometric curves of different film batches were evaluated in portrait and landscape scan mode. Between various batches important variations in sensitometric curve were observed. Energy dependence of the film is negligible, while a slight variation in dose response is observed for very large angles between film surface and incident photon beam. Improved accuracy in absolute dose detection can be obtained by repetition of a film measurement to tackle at least the inherent presence of film inhomogeneous construction. We state that the overall uncertainty is random in absolute EBT film dose detection and of the order of 1.3% (1 SD) under the condition that the film is scanned in a limited centered area on the scanner and at least two films have been applied. At last we advise to check a new film batch on its characteristics compared to available information, before using that batch for absolute dose measurements. PMID- 18383693 TI - Multifractality, sample entropy, and wavelet analyses for age-related changes in the peripheral cardiovascular system: preliminary results. AB - Using signal processing measures we evaluate the effect of aging on the peripheral cardiovascular system. Laser Doppler flowmetry (LDF) signals, reflecting the microvascular perfusion, are recorded on the forearm of 27 healthy subjects between 20-30, 40-50, or 60-70 years old. Wavelet-based representations, Holder exponents, and sample entropy values are computed for each time series. The results indicate a possible modification of the peripheral cardiovascular system with aging. Thus, the endothelial-related metabolic activity decreases, but not significantly, with aging. Furthermore, LDF signals are more monofractal for elderly subjects than for young people for whom LDF signals are weakly multifractal: the average range of Holder exponents computed with a parametric generalized quadratic variation based estimation method is 0.13 for subjects between 20 and 30 years old and 0.06 for subjects between 60 and 70 years old. Moreover, the average mean sample entropy value of LDF signals slightly decreases with age: it is 1.34 for subjects between 20 and 30 years old and 1.19 for subjects between 60 and 70 years old. Our results could assist in gaining knowledge on the relationship between microvascular system status and age and could also lead to a more accurate age-related nonlinear modeling. PMID- 18383694 TI - Spatial resolution properties in cone beam CT: a simulation study. AB - This work is intended to investigate the spatial resolution properties in cone beam CT by estimating the point spread functions (PSFs) in the reconstructed 3D images through simulation. The point objects were modeled as 3D delta functions. Their projections onto the detector plane were analytically derived and blurred with 2D PSFs estimated and used to represent the detector and focal spot blurring effects. The 2D PSF for detector blurring was computed from the line spread function measured for a typical a-Si/CsI flat panel detector used for general radiography. The focal spot blurring effect was simulated for an x-ray source with a nominal focal spot size of 0.6 mm and 1.33 x magnification at the rotating center. Projection images were computed and sampled with an interval significantly smaller than the detector pixel size to avoid aliasing. Images were reconstructed using the Feldkamp algorithm with the five different filter functions. Reconstructed PSFs were plotted and analyzed to investigate the effects of detector blurring alone, focal spot blurring alone, or a combination of the two on the PSFs and their variations with the radial distance and z-level. Effects of binning and reconstruction filters were also studied. Our results show that the PSFs due to detector blurring are largely symmetric and vary little with the locations of the point objects. With focal spot blurring only or added to detector blurring, the PSFs along the rotation axis were largely symmetric but became increasingly asymmetric as the point objects were moved away from the rotation axis. The PSFs were found to become wider in the axial (anode to cathode) direction as the objects were moved toward the cathode side. The 3D PSFs may be approximated by an ellipsoid with three different axial lengths. They were found to point upright along the rotating axis but tilt toward the rotating axis as the point object was moved away from the axis. PMID- 18383695 TI - Geometrical model-based segmentation of the organs of sight on CT images. AB - Segmentation of organs of sight such as the eyeballs, lenses, and optic nerves is a time consuming task for clinicians. The small size of the organs and the similar density of the surrounding tissues make the segmentation difficult. We developed a new algorithm to segment these organs with minimal user interaction. The algorithm needs only three seed points to fit an initial geometrical model to start an effective segmentation. The clinical evaluation shows that the output of our method is useful in clinical practice. PMID- 18383696 TI - Determination of dosimetric perturbations caused by aneurysm clip in stereotactic radiosurgery using gel phantoms and EBT-Gafchromic films. AB - Some radiotherapy patients are treated with titanium surgical aneurysm clips in the radiation field. This is of particular importance for stereotactic radiosurgery brain treatments, where the length of the blade of the clip may be comparable to the size of the radiation field. This study seeks to determine the extent of the dosimetric effects caused by surgical clips in stereotactic radiosurgery, using polyacrylamide gel phantoms and EBT type Gafchromic films. Using gel phantoms scanned with magnetic resonance imaging scanner, dose enhancement of around 20% was noted at distances less than 2 mm away from the clip surface. Gafchromic films showed about 6% variations in the dose up to few millimeters from the clip. These experimental results confirmed results predicted by Monte Carlo simulation techniques for higher density material surgical clips such as lead and platinum. Moreover, these experimental measurements clearly indicate dose reduction due to radiation attenuation behind the clip of about 4%. PMID- 18383697 TI - Inter fractional variability of breathing phase definition as determined by fiducial location. AB - Reconstruction of four-dimensional (4D) imaging typically requires an externally measurable surrogate to represent the real-time relative phase of respiration. A common method is to use a reflective marker on the external surface of the patient which moves with respiration and can be tracked in real time. The location of the marker is often chosen to maximize the observable motion, though this location may not be at the region of interest. We evaluate the importance of infrared (IR) marker placement location on breathing phase definition for the purpose of respiratory gating and 4D computed tomography (CT) image reconstruction. Data were collected for ten patients enrolled on an approved IRB protocol. Real-time position data were collected during CT imaging and daily treatments for two external IR reflective markers: one placed near the xyphoid and another at the approximate location of the treatment isocenter. Motion traces from the markers were compared using cross-correlation coefficient and by estimating the relative respiratory phase, based on either marker, as would be used for 4D-CT reconstruction. Cross-correlation analysis revealed differences in the motion waveform, as well as phase differences, both of which were variable between patients as well as day to day for the same patient. Estimated relative phases from each marker were compared by the percentage amount of time the estimated phase for each marker was different, binned based on increments of 10% of a full cycle. For all collected data combined, the frequency with which breathing phase mismatch led to different bin allocation in steps of 10% was as follows: T0%-10% = 65.1%, T10%-20% = 25.3%, T20%-30% = 7.8%, T30%-40% = 1.5% and T40%-50% = 0.4%. Based on ten images per cycle, this indicates that 4D reconstructions would be influenced, depending on which marker was used, by at least 1 bin 34.9% of the time. This number was noticeably higher for some patients; the maximum was 71% of the time for one patient of ten. In conclusion, the respiratory amplitude and relative phase depend significantly on the location of the IR marker used to monitor respiration. For some patients the xyphoid and isocentric markers may be completely out of phase. More importantly, this relationship varies day to day, suggesting that a single marker may be inadequate for the purposes of respiratory gating. PMID- 18383698 TI - Measurement of temporal resolution in dual source CT. AB - This work assesses the temporal resolution of dual-source computed tomography (CT) in a visually intuitive manner. Exploiting the principles of ring artifact creation, a phantom containing a highly attenuating delta function signal (a wire) was rotated at the same speed as the gantry, creating a partial ring artifact where the angular extent of the artifact provides a direct indication of the temporal resolution. A demonstration of the effect of the evaluated 165 and 83 ms nominal temporal resolutions on motion artifacts in cardiac CT is provided using patient data. PMID- 18383699 TI - Measurement of superficial dose from a static tomotherapy beam. AB - Superficial doses were measured for static TomoTherapy Hi-Art beams for normal and oblique incidence. Dose was measured at depths < or = 2 cm along the central axis of 40 x 5 cm2 and 40 x 2.5 cm2 beams at normal incidence for source to detector distances (SDDs) of 55, 70, and 85 cm. Measurements were also made at depths normal to the phantom surface for the same beams at oblique angles of 30 degrees, 45 degrees, 60 degrees, 75 degrees, and 83 degrees from the normal. Data were collected with a Gammex/RMI model 449 parallel-plate chamber embedded in a solid water phantom and with LiF thermoluminescent dosimeters (TLDs) in the form of powder. For comparison, measurements were made on a conventional 6 MV beam (Varian Clinac 2100C) at normal incidence and at an oblique angle of 60 degrees from the normal. TomoTherapy surface dose varied with the distance from the source and the angle of incidence. For normal incidence, surface dose increased from 0.16 to 0.43 cGy/MU as the distance from the source decreased from 85 to 55 cm for the 40 x 5 cm2 field and increased from 0.12 to 0.32 cGy/MU for the 40 x 2.5 cm2 field. As the angle of incidence increased from 0 degrees to 83 degrees, surface dose increased from 0.24 to 0.63 cGy/MU for the 40 x 5 cm2 field and from 0.18 to 0.58 cGy/MU for the 40 x 2.5 cm2 field. For normal incidence at 55 cm SDD, the surface dose relative to the dose at d(max) for the 40 x 5 cm2 TomoTherapy Hi-Art beam was 31% less than that from a conventional, flattening filter based linear accelerator. These data should prove useful in accessing the accuracy of the TomoTherapy treatment planning system to predict the dose at superficial depths for a static beam delivery. PMID- 18383700 TI - Evaluation of deformable registration of patient lung 4DCT with subanatomical region segmentations. AB - Deformable registration is needed for a variety of tasks in establishing the voxel correspondence between respiratory phases. Most registration algorithms assume or imply that the deformation field is smooth and continuous everywhere. However, the lungs are contained within closed invaginated sacs called pleurae and are allowed to slide almost independently along the chest wall. This sliding motion is characterized by a discontinuous vector field, which cannot be generated using standard deformable registration methods. The authors have developed a registration method that can create discontinuous vector fields at the boundaries of anatomical subregions. Registration is performed independently on each subregion, with a boundary-matching penalty used to prevent gaps. This method was implemented and tested using both the B-spline and Demons registration algorithms in the Insight Segmentation and Registration Toolkit. The authors have validated this method on four patient 4DCT data sets for registration of the end inhalation and end-exhalation volumes. Multiple experts identified homologous points in the lungs and along the ribs in the two respiratory phases. Statistical analyses of the mismatch of the homologous points before and after registration demonstrated improved overall accuracy for both algorithms. PMID- 18383701 TI - Mean position tracking of respiratory motion. AB - Modeling and predicting tumor motion caused by respiration is challenging due to temporal variations in breathing patterns. Treatment approaches such as gating or adaptive bed adjustment/ alignment may not require full knowledge of instantaneous position, but might benefit from tracking the general trend of the motion. One simple method for tracking mean tumor position is to apply moving average filters with window sizes corresponding to the breathing periods. Yet respiratory motion is only semiperiodic, so such methods require reliable phase estimation, which is difficult in the presence of noise. This article describes a robust method to track the mean position of respiratory motion without explicitly estimating instantaneous phase. We form a state vector from the respiration signal values at the current instant and at a previous time, and fit an ellipse model to training data. Ellipse eccentricity and orientation potentially capture hysteresis in respiratory motion. Furthermore, we provide two recursive online algorithms for real time mean position tracking: a windowed version with an adaptive window size and another one with temporal discounting. We test the proposed method with simulated breathing traces, as well as with real time displacement (RPM, Varian) signals. Estimation traces are compared with retrospectively generated moving average results to illustrate the performance of the proposed approach. PMID- 18383702 TI - Dynamic chemical shift imaging for image-guided thermal therapy: analysis of feasibility and potential. AB - A fast chemical shift imaging (CSI) technique based on a multiple gradient recalled acquisition using a small number of echoes with intentional aliasing of the reference lipid peak is studied to determine its feasibility for temperature monitoring. Simulations were implemented to find parameters where the lipid and water peaks can be measured using a Fourier-based peak fitting approach as well as using an innovative autoregressive moving average technique. A phantom consisting of 50% mayonnaise/50% lemon juice was calibrated to temperature and compared to literature values. A porcine kidney was treated ex vivo with an external laser and imaged with the CSI technique with comparisons to temperature readings from a fluoroptic monitoring system and complex phase difference (CPD) calculations. To demonstrate the technique in vivo, a Balb/c mouse with a CT26 xenograft in the subcutaneous lower back was treated using gold-coated, silica core nanoshells heated with an 808 nm interstitial laser. Compared to standard CPD techniques using a two-dimensional fast spoiled gradient recalled echo, this technique maintains spatiotemporal resolution, has high signal-to-noise ratio and accuracy over a wide range of T2* tissue values, can separate water and lipid signals, and additionally can use the lipid peak, when present, as an internal reference. PMID- 18383703 TI - [Ten year jubilee of the journal "Advances in Gerontology"]. AB - The article presents the report of the editorial board of the journal "Advances in Gerontology" devoted to 10th anniversary since the first publication. Analysis of character of the articles printed during last 5 years has been given, including their distribution by geography, departmental membership, and science themes. Geographic widening of the publications, increasing of number of institutions delivering the articles and of number of articles as well as number of the authors has been shown. These showings are the evidence of the fact that gerontology attracts more and more attention of the specialists. PMID- 18383704 TI - [Participation of apical dominance in processes of oncogenesis and old age involution: a hypothesis]. AB - Apical dominance (the phenomenon known in plants) underlies pathological morphogenetic processes occurring in animals and human in oncogenesis and senescence. PMID- 18383705 TI - [Menopause--a key aspect of aging: role of the pineal gland]. AB - To estimate the sign and measure of changes of the pineal gland volume in perimenopausal period a magnetic-resonance tomography studies of the head of men (n=72) and women (n=74) of 38-61-year old were organized, in group divided into two subgroups with a space of 2 years. For estimation of possible dependency of the pineal gland volume on the reproductive cycle phase the similar study was organized in groups of women of the first (n=34) and second mature age (n=41) with regular cycle who were divided into subgroups by the phase of the cycle fixed retrospectively by the basal temperature graphs. The valid reduction of the pineal gland volume was registered in women of ovulate phase as well as during perimenopausal period. In context of literary data concerning recurrence and age dynamic of melatonin production by the pineal gland participation of pineal gland in "shutdown" of reproductive functions in women is supposed. PMID- 18383706 TI - [Morphofunctional status of gonadotropic cells of the adenohypophysis at early stages of age involution]. AB - Action of alpha-tocopherol, emoxipinum on functional status of gonadotropic cells was investigated at deficiency of sexual hormones in male and female rats of Wistar line. The alpha-tocopherol slows down aging of gonadotropic cells after gonadectomy. PMID- 18383707 TI - [Apoptosis regulation in hypothalamic neurosecretory cells of HER2/neu transgenic mice in ontogenesis]. AB - Tyrosine kinase receptor HER2/neu plays an important role in a number of processes including carcinogenesis. The oncogenic characteristics of HER2/neu are associated with its ability to affect a variety of apoptotic pathways creating, this way, an antiapoptotic environment in the cells overexpressing this protein. The aim of our work was to investigate the features of apoptosis regulation in hypothalamic neurosecretory cells of HER2/neu transgenic mice in aging. We detected the apoptosis protein expression (Bax, c-Raf) in comparison with apoptosis level and functional activity (vasopressin concentration) in neuroendocrine system. Besides, we studied the level of 17beta-estradiol in blood plasma. 17beta-estradiol is one of possible antiapoptotic factors in neurons. We show that the apoptosis of neuroendocrine cells increases in aged wild type mice, but not in HER2/neu ones. Recently we obtained that the mechanism of apoptosis suppression in transgenic mice is the block of p53-dependent apoptosis cascade, and it is the cause of caspadse-8 decrease and dysregulation of Bcl-2 and Mcl-1 antiapoptotic protein synthesis. In this study it has been shown that Bax concentration decreases and c-Raf-1 expression does not change. 17beta-estradiol does not decrease in plasma of aged transgenic mice and it is the factor, which can play a positive role in neuroendocrine cells survival. Besides, the vasopressin synthesis increases in young and old HER2 mice. These facts result in the increased survival of neurosecretory cells in old transgenic mice. PMID- 18383708 TI - [Recessive lethal mutation in X-cromosome locus 6D1-7A2 in heterozygote leads to life span increasing in Drosophila melanogaster]. AB - The results of demographic analysis of survival of different Drosophila lines have shown that recessive lethal mutation located in X-chromosome locus 6D1-7A2, as compared with wild type strain, in heterozygote leads to aging speed deceleration (decreasing of mortality trajectory slope), median (20%) and maximal (15-20%) life span increasing. Further investigations (complementation analysis with duplicated loci) will enable to discover, whether it is unknown life span gene. PMID- 18383709 TI - [Effect of light regimens and melatonin on the homeostasis, life span and development of spontaneous tumors in female rats]. AB - Effect of different light regimens (standard--12 hours of light/12 hours of darkness, natural lightning of the North-West of Russia and constant illumination) and melatonin on the homeostasis, life span parameters and spontaneous tumor development of female rats was studied. Animals at conditions of constant or natural light regimens revealed accelerated aging of organism, decrease of mean (by 13.5% and 25.5% accordingly) and the maximal life span (by 9 and 7 months, accordingly) and faster spontaneous tumor development as compared with the animals kept at the standard light regimen. Melatonin given in nocturnal drinking water (10 mg/l) prevented the adverse effect of the natural light regimen on the homeostasis, life span and tumor development. Melatonin also decreased spontaneous tumor development, first of all, mammary and uterine neoplasm, in the rats kept at the constant illumination. PMID- 18383710 TI - [Premature aging of an organism and characteristics of its manifestation in remote period after low dose irradiation]. AB - In this study 58 participants of the liquidation of the consequences of Chernobyl accident in 1986-1987 were investigated. All the patients complain of constant headaches, disorders of memory, general weakness, rapid fatigability, decreased sexual drive, emotional instability etc. The complex (comprehensive) modern methods of investigation were used to carry out the objective assessment of presented complains and of character of the central nervous system damage: complex computer quantitative analysis of mental capacity; analysis of personality traits by using the MMPI test; single photon emission tomography (with the drug of Ceretec); X-ray computer tomography; magnetic resonance computer tomography. The experimental study with examination of primates who were exposured in sum dose 1 Gy (by drop method) was carried out, too. The results of complex investigation of participants of liquidation of Chernobyl accident consequences enable to postulate the formation of premature aging of an organism in these persons. Data of the experimental study of primates irradiated in dose 1 Gy revealed formation of the brain atrophy in the remote period after low dose radiation exposure. PMID- 18383711 TI - [Effect of Earth magnetic field on circadian rhythm of total antioxidant capacity of human saliva in the North]. AB - In the inhabitants of the North during increase of geomagnetic activity and during magnetic calm the decrease of amplitude of circadian rhythm of total antioxidant capacity of saliva is observed. The most favorable conditions to display the circadian rhythm are observed at Kp from 0,5 up to 2. The long residing in the North is connected to influence of irregularly varying geomagnetic activity causing disturbance of function of circadian and antioxidant systems that, probably, is one of the reasons of acceleration of process of aging at northerner and of higher risk of occurrence in them the age associated diseases. PMID- 18383712 TI - [The effect of pineal peptide preparations on proliferative activity in organotypic culture of the preoptic hypothalamus area]. AB - In investigations carried out with organotypic culture of mediobasal preoptic area (MPA) of hypothalamus it was established that pineal peptide preparations epitalon (2 ng/ml) and epithalamin (100 ng/ml) stimulate the development of proliferative activity of explants in 3 month and 24 months female rats. It has been shown that epithalamin is more effective in young rats in comparison with old animals, but epitalon as well as epithalamin have almost the same less pronounced inducing effect on growth zone in MPA explants from young and old animals. This effect is tissue specific and could be dependent on inhibition of apoptosis. PMID- 18383713 TI - [Effects of ligfol after radiation-mercury exposure in rats of various age]. AB - The study was carried out with the young (4-5 months) and elderly (15-18 months) female and male rats at the experiment start. The mercury salt was administered into animals with drinking water in 1 mcg/l concentration during 3 months before and 1 month after single X-irradiation with 25 cGy dose. The genotoxic effects in leukocytes were evaluated on the base of the content and structure DNA alterations in 24 hrs and in 1 mth after irradiation. The alaninaminotransferase activity in serum was determined to evaluate functional status of liver. A protector intramuscular injection of Ligfol preparation with the 250 mg/kg dose in 24 hrs before irradiation resulted in increase of the blood leukocyte amount, more expressed in 1 mth after irradiation. The standardizing effect of preparation on the leukocyte DNA content is manifested in 1 mth after combined exposure on rats within young male and elderly female. At the same time, Ligfol showed some restoring effect on the alterations of leukocyte supercoiled DNA structure in 1 mth after irradiation within elderly female rats. The preparation showed the standardizing effect on the liver cells in young males injured by radiation-mercury exposures with low doses, however Ligfol had not influence practically on the destruction degree of hepatocytes after similar injury within female young and elderly rats. So, results of the intramuscular Ligfol injection is manifested at least during 1-2 mths; the young rats of both gender and elderly females showed the most sensitivity to the radiation-mercury exposure with low doses and to the Ligfol, according to the studied indexes. PMID- 18383714 TI - [The role of melatonin in regulation of gonadal function and its use in the treatment of pathological climax symptoms]. AB - Pathological current of climacterium leads to significant decrease in quality of life of women. Hormone therapy has a number of contra-indications. With the purpose of optimization of treatment of patients with a climacteric syndrome search of new and more perfect approaches to therapy of climacteric frustration is necessary. Melatonin production in a human body decreases with the years and its production in old age makes only half from its level. Age decrease of melatonin synthesis leads to progressive decrease of woman fertility. In this connection the research of a level of melatonin in women with a climacteric syndrome depending on current of disease was carried out. The opportunity to use the Melaxen in treatment of symptoms of pathological climacterium both in the form of monotherapy, and in combination with replacement hormone therapy was estimated. The greatest efficiency of application of Melaxen in combination with hormone replacement therapy has been established. PMID- 18383715 TI - The principles of chemotherapy of colorectal cancer in elderly. AB - The prevalence of colorectal cancer (CRC) increases significantly with age, with 40% of patients in Europe being older than 74 years of age at the time of initial diagnosis. The individualized management of the older-aged patient with cancer is based on the answers to the following questions: 1) will the patient die of cancer or with cancer; 2) will the patient suffer cancer-related morbidity; and 3) is the patient able to handle the toxicity of treatment? More than chronological age, the following parameters are important when elderly patients are to be treated with antineoplastic agents: general condition, liver function, kidney function and bone marrow status. Frail elderly with malignant disease should not be treated with cytostatic therapy. In the case of fit elderly, the standard chemotherapy (i.e. FOLFOX) regimen could be administered. In elderly ineligible for combination chemotherapy, the capecitabine used orally, as a single-agent therapy, is an important therapeutic option for colorectal cancer. PMID- 18383716 TI - [Postoperative complications of gallstone disease in elderly and aged patients]. AB - Here we have analyzed the results of surgical treatment of gallstone disease and its complications in 1208 senior-age patients. In 875 patients planned operations were performed, whereas in 333 patients the interventions were urgent. Post operational complications occurred in 5.2% of cases, and lethality amounted to 4.1%. The complications and post operational lethality depended on how traumatic and urgent the intervention was. Diagnostics of intraperitoneal complications was based on instrumental examination methods, with primary significance assigned to laparoscopy. In 21 patients early post operational intraperitoneal complications were treated with the use of mini-invasive methods. Guidelines for surgical treatment of gallstone disease and its complications in senior patients have been worked out. PMID- 18383717 TI - [Some problems of interrelations between ischemic stroke and cranial-cerebral trauma in elderly and aged patients]. AB - The matters under studies were interrelations between cranial-cerebral trauma and ischemic stroke in old patients. Cranial-cerebral trauma took place before ischemic stroke in 80 cases. Ischemic stroke took place before cranial-cerebral trauma in 174 cases. There was determined predominance of vessels neurologycal simptoms in combination with cranial-cerebral trauma and ischemic stroke. Some symptoms of cranial-cerebral trauma predominate only in acute traumatic period. But the vessels neurological symptoms become prevailing again after successful treatment in connection with cranial-cerebral trauma. This determines that cerebral vessels pathology always predominates as regards cerebral traumatic pathology in old patients. PMID- 18383718 TI - [The application of the selective proximal vagotomy by the method of chemical denervation in treatment of patients of elderly and senile age suffering from the perforating pyloroduodenal ulcers]. AB - The article presents the results of using the vagotomy of an acid-producing zone of a stomach by the method of chemical denervation. Subjects to the operation were 20 Wistar line male rats and 22 patients with perforating pyloroduodenal ulcers accompanied by cardio pathology. The simplicity, availability and low traumatism of the technique, in view of the low percent of a relapse of the pyloroduodenal ulcers, enable to recommend it for the treatment for the elderly patients with the heavy cardio pathology. PMID- 18383719 TI - [Specific features, problems, and prospects of myocardial revascularization in elderly and aged patients]. AB - Last years the progress in development and improvement of coronary surgery and angioplasty, with shunting with patients of elderly and senile age, has been observed in the Russian Federation. This is connected, in the first place, with the progress in timely diagnostics of coronary artery disease, development of surgical equipment, anaesthesiology and resuscitation, introduction of new methods of intra-operative protection of myocardium, development and active use of modern methods of treatment of patients before and after the operation. However despite the versatility of existing scientific schools and literary sources covering this problem, the discussions on indications and contraindications to active surgical treatment of coronary artery disease with the patients of the given age group still continue. A great number of specialists in the field of cardiology and cardiosurgery do not tackle the solution of these problems as based on age-specific morphological and functional changes of the cardiovascular system. PMID- 18383720 TI - [Effect of osteoinductors on osseous tissue reparation in ageing]. AB - Ageing of population determines the ever growing need for dental health service to older patients, which stipulates the necessity of the search for new materials with osteoplastic and osteoinductive properties. Experimental study on old animals showed that osseous defects of the lower jaw, filled with granules of GAP 99 osteoinductor, were completely replaced with new osseous tissue by the 90th day of the experiment. PMID- 18383721 TI - [The factors of oxidative stress in neurodegenerative diseases (vascular dementia, Alzheimer disease)]. AB - We investigated the state of oxidative stress in aging people suffering from vascular dementia and Alzheimer's disease. We have examined the oxidative modification of plasma's proteins, the activity of enzyme antioxidative defense (superoxide dismutase and catalase) and the state of components of blood glutathione system. Aged patients with neurodegenerative diseases show decrease of metal-catalysed oxidative modification proteins, increase of superoxide dismutase and disrupt of balance enzymatic antioxidant and glutathione system in blood in comparison with aging people who have no psychoorganic disorders. The high activity of superoxide dismutase in patients with Alzheimer's disease has been discovered. We have shown the joint change of separate component oxidative stress with the degree of mental disorders. Oxidative stress is believed to play a role in the pathogenesis of different neurodegenerative diseases in aged patients. PMID- 18383722 TI - [Medical and psychological prevention of stress-induced premature aging]. AB - We studied the efficiency of the method complex for psychotherapy (psychocorrection) of those who suffer from post-traumatic stress disorders (PTSD) with the purpose of prevention and reduction of psychovegetative disorders and aging pace which would prevent from early disability and death. We studied 82 male patients suffering from post-traumatic stress disorder aged from 22 to 35 who endured combat psychologic traumatic experience. In our study we used 4 method approaches enabling to assess a functional state of an organism sequentially and fully. Those included: 1) a clinical psychopathological approach; 2) an experimental psychological approach; 3) a spectral analysis of the heart rate variability; 4) assessment of biological age dimensions. It was ascertained that PTSD can be seen as one of the factors increasing BA (biological age) and aging pace of people who experienced extreme situations. Our study proved that combined methods of psychotherapeutic rehabilitation also normalize parameters of vegetative heart rate regulation circuit. It is shown that positive changes in psychovegetative sphere achieved in people suffering from PTSD decrease an organism's aging pace. PMID- 18383723 TI - [Social and psychological aspects of longevity]. AB - The fact that longevity is unclaimed in the society is discussed. In this connection two social models of ability to live are offered to consideration. The first one is a "being" model determined by the mass consciousness of social space. The second one is a "cognition" model being formed meaningly by a person on the basis of the own interpretation of the universe. Their influence on the process of longevity is analyzed. PMID- 18383724 TI - [Socio-cultural model of gerontosocial work for improving quality of life of elderly people]. AB - Scientific and theoretical approaches to gerontosocial work, perspective trends of its development in the Russian society: socio-cultural model, resource based gerontosocial work, two-level training system for specialists on gerontology, gerontological education have been discussed. The goals and levels of education in gerontology in the modern society have been considered. The introduction of the socio-cultural model into the practical work with the elderly people in accord with the Madrid Action Plan on Ageing has been proved. The perspective trends of improving quality of life of the elderly people within the framework of the gerontosocial policy have been suggested. PMID- 18383725 TI - [Urgent problems of elderly in working and family environment (according to opinion poll 2004-2006)]. AB - Problems considered concerned of elderly in a family (with young community especially), with authorities, on the female loneliness, and also on the professional activity. PMID- 18383726 TI - [Caspase-3 activity in the rat hippocampal slices reflects changes in synaptic plasticity]. AB - Considering the involvement of caspase-3 in neuronal plasticity, we studied caspase-3 activity in the rat hippocampal slices, and electrophysiological characteristics of extracellular responses to paired-pulse stimulation of Schaffer's collaterals in the CA1 subfield of hippocampus. Caspase-3 activity was measured after electrophysiological recording in each slice separately. Maximal caspase-3 activity was observed in the slices with low responsiveness to single afferent stimulation indicative of decreased efficacy of interneuronal interaction. This phenomenon is unrelated to depression of neuronal excitability since paired-pulse stimulation increases the synaptic efficacy to second stimulus thus restoring population spike amplitudes to normal values. In "damaged" slices with impaired spike generation up to disappearing spikes to both stimuli, caspase 3 activity was close to the normal level of the "healthy" slices. The activity of another proteinase, cathepsin B, was increased in the "damaged" slices, no correlation with the modifications of electrophysiological indices being detected. Our data suggest that high caspase-3 activity in hippocampal slices is involved in maintenance of synaptic plasticity but not necessarily related to apoptosis. PMID- 18383727 TI - [Long-term disorders in the rat behaviour induced with introduction of the tumour necrosis factor in early postnatal ontogenesis]. AB - Anti-inflammatory cytokin the tumour necrosis factor alpha (TNFalpha) is one of the main mediators of neuroimmune interactions. The TNFalpha production increase in the cells of neural and immune systems occurs in infectious diseases, injuries, ischemia, hypoxia. In this study, the TNFalpha revealed its role in formation of behaviour in early postnatal ontogenesis. The rats with the TNFalpha administered during their 3 weeks of life differed from the control those by a change in the orienting behaviour and enhanced anxiety. The role of the anti inflammatory cytokins in the brain during different stages of ontogenesis is discussed. PMID- 18383728 TI - [The causes of intragroup aggression in rhesus monkeys]. AB - The causes of aggression in coral monkeys have been considered taking into account the composition of conflicting pairs and the forms of aggression. It was established that more than 85% of the aggressive interactions occurring on the background of a group calmness, were noted in a context of avoidance of closeness with an undesirable partner, demonstration of hierarchical status, competition for closeness to a preferable partner, competition for the resources, the defense of an infant, and complications during the play and sexual herding. PMID- 18383729 TI - [The effect of synthetic fragment 65-76 of monocyte chemiattractant protein-1 (MCP-1) on neointima growth after carotid artery balloon injury in rats]. AB - Influence of synthetic fragment 65-76 of monocyte chemoattractant protein-1 (MCP 1) (peptide X) on development of neointima after balloon injury of carotid artery was investigated. Peptide X was introduced intramuscularly, 33 pg/kg, daily during 28 days after balloon injury. In days 4 and 7 after intervention, in animals receiving peptide X in comparison with control animals a substantial decrease of neointimal growth was observed. On 14 and 28 days there, was no significant difference in neointima development in rats with and without peptide treatment. Injections of peptide X did not after the C-reactive protein concentration, leukocyte number and lymphocyte subpopulations in peripheral blood. Peptide X treatment along with traditional therapy may be effective in preventing restenosis after angioplasty. PMID- 18383730 TI - [Restoring effect of noradrenalne on diminished neurogenic vasoreactivity]. AB - The effect of 0.01-1.0 microM noradrenali on response to electrical field stimulation (EFS) of the juvenile rat tail artery segment was studied. Noradrenali was shown to potentiate the EFS-evoked constriction decreased in the course of experiments or in the acidic solution (pH 6.6) and this potentiation was proportional to the extent of the preceding decrease of the constriction. The more decreased was the EFS-evoked constriction the higher was the noradrenali concentration which produced the maximal potentiation and the wider was the potentiative noradrenali concentration range. The potentiative effect of noradrenali was not prevented by the NO synthase inhibitor NG-nitro-L-arginine. The results suggest that noradrenali can restore the diminished neurogenic reactivity of blood vessels, and this effect is not connected with the change in the NO production. PMID- 18383731 TI - [Leukocyte-endothelium interaction in pial arterioles and venules following cerebral ischaemia of rat]. AB - With the help of contact optic system leukocytes interaction to endothelium of both pial arterioles and venules was investigated during cerebral ischaemia caused by bilateral occlusion of carotids, in vivo. The data received on 40 arterioles and 30 venules (diameter under 40 microns) of pia matter of Wistar rats (n = 7) under ischemic conditions following 5 hours up to respiratory arrest were analyzed. In this experiment, significant differences in adhesiveness of leukocytes to endothelium of arterial and venous microvessels during hypoxia development were shown. PMID- 18383732 TI - [The effect of surgery with cardiopulmonary bypass on oxygen transport in elderly patients with ischemic heart disease]. AB - In a clinical trial, 103 patients undergoing coronary artery bypass grafting from May, 1999, to December, 2001 with hypothermic cardiopulmonary bypass (CPB) were retrospectively assigned to one of two groups: group I (n = 45)--patients 65 years of age and older (68.0 +/- 0.5), group II (n = 58)--patients 45-50 years of age (48.2 +/- 0.2). The following parameters were recorded: haemodynamic--with thermodilution method (SMU--612, Hellige), blood gases (OMNI-6, Austria). Measurements were performed 7 times: (1) before surgery, (2) before CPB, (3) after CPB, (4, 5, 6, 7)--1, 3, 9, 12 hours after surgery. Indexes of oxygen delivery and consumption, oxygen utilization coefficient and anion gap were calculated. Obtained results were statistically analyzed using appropriate t-test and chi2-test for categorical variables. Data are expressed as mean +/- SE. There were no significant differences between the groups in all stages of examination, p > 0.05. In elderly group both oxygen delivery and consumption were lower then in younger one, thus the coefficient of oxygen utilization did not differ between groups. Therefore the surgery with CPB seems to be inrelated to adverse changes on oxygen transport in elderly patients, and its dynamic was similar in patients over 65 years of age and younger group. PMID- 18383733 TI - [Functional and metabolic changes of healthy volunteers after cold exposure and administration of meteoadaptogen trekrezan]. AB - The effect of cold exposure (-10 degrees C, air speed--2.5 m/sec, 40 minutes) on physical activity, cognitive processes and metabolic status of 75 volunteers, healthy men of 20-24, was studied in termobarocomplex Tabaj (Japan). Cold exposure reduced physical and cognitive activity, the activity of kreatine phosphokinase, superoxide dismutase, the levels of redox glutation and pyruvate. Preliminary administration of adaptogenic drug trekrezan 0.2 g prior to cold exposure normalized the indexes studied of physical activity and metabolic status. It is suggested that trekrezan can be used as a meteoadaptogenic drug for rapid and effective adaptation to cold exposure of environment. PMID- 18383734 TI - [Artificial vestibular feedback in conditions of modified body scheme]. AB - Subjects standing in darkness on the rigid support, kept a vertical posture which was destabilized by vibration of the Achilles tendons. To create a feedback on the vestibular input, transmastoidal bipolar galvanic stimulation was used. Changes of current in the feedback contour looked as linear function considering amplitude and velocity of the subject's head displacements in reference to the vertical. To change the body scheme we used some posture configurations: turning of the head in relation to the trunk; turning of the trunk with the head fixed; joint turning of the head and trunk. As a result of these configurations, the head could be turned approximately at right angle in relation to the feet. In addition turning of one foot at right angle in relation to the other foot was used. Artificial feedback reduces body fluctuations caused by vibration only in the vertical plane which passes through interaural axis of the head. The authors assume that directional changes of vestibulo-motor responses and results of application of artificial feedback during changes of orientation of the head in relation to the feet can be connected to change of ensembles of vestibular hair cells, which signals dominate in responses of vestibulo-spinal neurones. PMID- 18383735 TI - [Forming of the visual cognitive structures in the monkey conditioned-reflex behaviour: the dependence on the sensory information]. AB - In monkeys, changes in size and shape of figures led to a significant decrease of correct solutions in training and a considerable increase of refusals from solution of tasks as well as the time of their motor response. The invariance of differentiation in this case was achieved after additional training. The data obtained show that, based on the stimulus sensory processing in conditioned reflex training, in the long-term memory some differentiating signs are formed: the cognitive structures (the functional neurophysiological mechanisms) maintaining the classification of visual images. With these structures, temporary conditioned connection will be established. Their formation will be determined by the type of sensory information and provided for by existence in the long-term memory of separate subsystems for spatial as well as non-spatial information. PMID- 18383736 TI - [Respiratory responses to microinjections of leptin into the solitary tract nucleus]. AB - Regulatory polypeptide leptin, apart from its well-known hypothalamic effects, stimulates ventilation. The present study on anaesthetised rats was undertaken to elucidate the respiratory effects of 10(-10)-10(-4) M leptin microinjected into the solitary tract nucleus, containing a high concentration of leptin receptors. Injections of 10(-8)-10(-4) M leptin induced dose-dependent increase in ventilation, tidal volume and electric activity of inspiratory muscles; 10(-6) M leptin additionally induced a short-term increase in respiratory frequency and a shortening of both inspiratory and expiratory duration. The respiratory responses to leptin is also characterised by appearance of sighs: deep and prolonged inspirations associated with an augmented burst in the activity of the inspiratory muscles and prolonged post-sigh inter-burst interval. The results taken together with evidence of high concentration of specific leptin ObRb receptor in the solitary tract nucleus suggest involvement of endogenous leptin in the control of breathing via dorsal structures of the respiratory center. PMID- 18383737 TI - [The age dynamic of hormonal levels in children living in different latitudes in European North]. AB - An investigation simultaneously of hormonal profile in children of 10-16 years living in different latitudes and climato-ecological conditions in Russian European North (in the border of polar and middle latitudes) was performed. The increased thyrotropin level was revealed in 35% of children in the south (61 degrees North) and in 32% of children in the North (64 degrees 30' North). The thyroid hormones (thyroxin, triiodothyronine) are at low normative bound. Higher Insulin levels were observed in 28% of children living in the southen region especially in 50% of prepubertal boys. In 20% of children, higher cortisol levels were revealed mainly at the final puberty (35% of children). PMID- 18383738 TI - [Uterus sympathetic nervous apparatus in sexual cycle dynamics in rats]. AB - Uterus sympathetic nervous apparatus in various phases of sexual cycle in rats was studied with the microspectral fluorescent method. The main part of nervous fibers in the form of perivascular plexus and terminals deviating from them are localized in myometrium. We suppose presence of reserve ("silent") nervous fibers in uterus which explains the inconstancy of their revealing in the organ. Catecholamines and serotonin are identified in varicose enlargements and intervaricose parts of nervous fibers by the way of microspectrofluorometry. Their concentration in probing places is characterized by high degree of linear correlation at all stages of sexual cycle. Maximal concentration of bioamines in most structural elements of uterus sympathetic nervous apparatus in detected in late diestrus. Stages of early and late estrus and proestrus are characterized by low levels of monoamines in adrenergic fibers. A number of reliable differences between bioamines saturation and spatial distribution density of nervous fibers in the neck and body of the womb are revealed. PMID- 18383739 TI - [Croatian guidelines for use of eicosapentaenoic acid and megestrol acetate in cancer cachexia syndrome]. AB - Among many oncological patients we can notice a substantial loss of body weight, fat and proteins with significant proinflammatory activity at the time of diagnosis. This wasting condition is well known as cancer cachexia syndrome. Anorexia is important part of this syndrome. Because cancer cachexia reduces tumor response to treatment and it is an indicator of poor prognosis, we need to start correcting these nutritional deficits at once. In the presence of cancer cachexia it is extremely difficult to achieve protein anabolism and stop the body wasting by standard nutritional formulas only. During the last few years, the use of eicosapentaenoic acid (EPA) and megestrol acetate (MA) as anticahectic agents has been tested. These guidelines are intended to give evidence-based recommendations for the use of eicosapentaenoic acid and megestrol acetate in cancer cachexia syndrome. These guidelines have been developed by interdisciplinary expert group of Croatian clinicians. Based on relevant literature, we have concluded that the use of metabolic modulators such as eicosapentaenoic acid and megestrol acetate for 8 weeks may help to improve nutritional status in cachectic patients. PMID- 18383740 TI - [Erlotinib in non-small cell lung cancer: experience of clinical hospital for lung diseases Jordanovac]. AB - Treatment with erlotinib, an inhibitor of the epidermal growth factor receptors, has significantly improved the overall survival rate and quality of life in patients with non-small cell lung cancer who had failed the standard first- or second-line chemotherapy. In Clinical hospital for lung diseases "Jordanovac" a total of 36 patients were treated with erlotinib. The response rate was as follows: in four patients (11%) complete response was achieved, in five (4%) partial response, while fourteen patients (39%) had stable disease. Hence, the evident clinical benefit of treatment with erlotinib was registered in 23 patients (64%) altogether. The treatment was well tolerated and it was not associated with significant toxicity. Our results confirm the antitumour efficacy of erlotinib and show a clear treatment benefit for patients with non-small cell lung cancer in our country. PMID- 18383741 TI - [Primary central nervous system lymphoma--a single center experience]. AB - Primary lymphoma of central nervous system (PCNSL) represents a distinct form of extranodal non-Hodgkin's lymphoma localized to central nervous system. We collected data retrospectively of 20 patients with PCNSL treated at Division of Hematology of UHC Zagreb in the last eight years. A total of 13 patients received high dose methotrexate (4 g/m2) while others received other chemotherapy regimens or radiotherapy only. Complete remission rate was 40% and estimated 2-years actuarial survival was 30%. There were no treatment related deaths or significant severe adverse events. Our results are satisfactory in patients younger than 60 years, especially if treated with high dose methotrexate. Older patients, who represent majority of the patients, have dismal prognosis irrespective of treatment modality. PMID- 18383742 TI - [Classical type of Fabry disease without angiokeratomas--a case report]. AB - A patient with classical type of Fabry disease is described. The appearance and character of neuropathic pain during hemodyalisis is described. Characteristic changes in head shape and changes of hands and fingers are the additional phenotypic characteristics of Fabry disease. Enzyme replacement therapy administered in the early phase of the disease could prevent disease complications and early patient's death. PMID- 18383743 TI - [The past and present status of "motor speech area": from classical modular "center" to nodal and supramodal component of multiple neuronal networks]. AB - In the classical, aphasiological (Wernicke-Lichtheim) model for neural basis of language, speech production was assigned to the "motor speech area" located in opercular and triangular part of the left inferior frontal gyrus (fronto opercular cortex, Broca's area) and conceptualized as a classical "modular centre" within a simple, language-specific neural network. However, recent neuroimaging studies on healthy volunteers have demonstrated a far more complex connection between brain and language: the left fronto-opercular cortex is just one of many nodal points in multiple and partially overlapping neural networks subserving language. In addition, the fronto-opercular cortex is not a simple motor centre, but part of supramodal (executive) frontal system with a number of language and non-language functions. We review these novel roles of the left fronto-opercular cortex, with focus on its functions which extend far beyond the scope of classical Wernicke-Lichtheim model. PMID- 18383744 TI - [Botulinum toxin type A and cholinergic system]. AB - Anaerobic bacterium Clostridium botulinum produces seven different serotypes of botulinum neurotoxins (A-G), which specifically act at the peripheral cholinergic nerve terminals blocking the release of acethylcholine. Primary site of action of botulinum toxin type A (BT-A) is neuromuscular end plate where it specifically cleaves SNAP-25, one of the proteins necessary for neuroegzocytosis. The consequence is long-lasting muscle paralysis. Although BT-A is one of the most potent toxins in nature, over the last 20 years, intramuscular injections of nanogram quantities of BT-A have been used to treat various conditions characterized by increased muscle contraction, like dystonias, spasticity related to cerebral palsy etc but also for autonomic nervous system disorders, like hyperhydrosis. Long duration of action (several months) after peripheral application is the most prominent feature of the toxin's action. Although the acute mechanism of action on neuromuscular junction is largely investigated, there are still some unknowns related to: the passage of BT-A through epithelial barriers, specific recognition of peripheral cholinergic neurons. The mechanism of long duration of action, which is the base of therapeutic use of BT-A, is poorly understood. PMID- 18383745 TI - [Nikola Tesla in medicine, too]. AB - Using primary and secondary sources we have shown in this paper the influence of Nikola Tesla's work on the field of medicine. The description of his experiments conduced within secondary-school education programs aimed to present the popularization of his work in Croatia. Although Tesla was dedicated primarily to physics and was not directly involved in biomedical research, his work significantly contributed to paving the way of medical physics particularly radiology and high-frequency electrotherapy. PMID- 18383746 TI - [Specific microbiological diagnosis of Entamoeba histolytica infection]. PMID- 18383747 TI - [New fluids for peritoneal dialysis]. PMID- 18383748 TI - [Ciprofloxacine and epilepsy]. PMID- 18383749 TI - [Vitamin D insufficiency in patients with chronic renal disease]. PMID- 18383750 TI - External monitor for buried free flaps in head and neck reconstructions. AB - Head and neck defects following oncological surgery must often be repaired with soft tissue and/or bone from other areas of the body. Significant loss of soft tissue requires flaps of sufficient bulk to adequately reconstruct the defect. Microvascular free tissue transfer is a good method for reconstructing even large defects following oncological surgery for head and neck cancer. Continuous post operative monitoring of the perfusion of a free flap is vitally important to achieve not only a favourable outcome but also to decrease morbidity. Microvascular thrombosis occurs in 4% of the flaps and the best chance for flap salvage is offered by the earliest possible revision of the microanastomosis. Use of buried flaps in head and neck reconstruction makes monitoring particularly difficult and exteriorization of a segment of the flap permits a direct visualization. An original technique is presented for harvesting forearm free flaps with a secondary monitor skin paddle to externally check the status of the paddle and, when modified, can also be used for fibula and rectus abdominis flap. PMID- 18383751 TI - Effectiveness of sulphur spa therapy with politzer in the treatment of rhinogenic deafness. AB - Several studies have focused on the usefulness of sulphur, radioactive and bromo iodine mineral waters in the treatment of chronic inflammatory lower and upper respiratory processes. The purpose of this study was to evaluate the tolerability, effectiveness and impact on quality of life of sulphur spa therapy with Politzer in subjects with chronic inflammatory processes responsible for the onset or persistence of rhinogenic deafness. The study was performed on 27 subjects (mean age 62 +/- 2.2 years, range: 28-88) with chronic catarrhalis otitis, chronic rhino-sinusitis and pharyngeal inflammation. These patients underwent 12 sessions of humid-hot inhalation, with vapour jet 20 cm from the face, at 38 degrees C for 10 min, followed by Politzer with sulphur sodium chloride bicarbonate alkaline mineral water from "Rosapepe" Spa, in Contursi (Salerno, Italy). Middle ear function and possible social recovery (based on Giaccai and Gardenghi guidelines) of the patients were assessed, at the beginning and end of the spa therapy. Results, at the end of this treatment, showed a significant (p < 0.05) increase in audiometric curves corresponding to the normal ventilation of the tympanic box (incidence of 24% before therapy and 33% thereafter) and a decrease in pathological curves. Moreover, a significant (p < 0.05) reduction in the percentage of auditory loss was recorded (N = 41; 19.7% +/ 2.5 --> 13.9% +/- 1.9) and improved hearing, at the frequencies required for daily activities: 500-1000 and 2000 Hz (31.1 dB +/- 1.7 --> 26.8 dB +/- 1.5). No adverse effects to the spa therapy were observed during the study. In conclusion, the results of this study are in agreement with data in the literature, demonstrating that associated spa therapy with Politzer and inhalation have a positive impact on the therapeutic strategy of chronic inflammatory processes, responsible for the onset or persistence of rhinogenic deafness, in order to enhance and combine with the already consolidated pharmacological approaches. PMID- 18383752 TI - Transtympanic steroids in refractory sudden hearing loss. Personal experience. AB - The treatment of choice for sudden sensorineural hearing loss is still lacking. Many drugs have been used over the years, with varying results and steroids have proven to be effective in clinical trials, albeit systemic administration is associated with untoward side-effects and cannot be used in all patients. The transtympanic approach presents two main advantages: first, it allows higher concentrations in the inner ear environment and, second, it minimizes systemic absorption. Aim of the present investigation was to establish the effectiveness of transtympanic steroid treatment for sudden sensorineural hearing loss in patients in whom conventional treatment had failed. For this purpose, a prospective, non-randomized study was designed to evaluate hearing improvement in sudden sensorineural hearing loss patients treated with transtympanic steroids. A solution of methyl-prednisolone and sodium bicarbonate was administered, via a transtympanic injection, in 10 patients. Hearing levels were evaluated before treatment and on days 1, 7 and 30, thereafter. Improvement in hearing was observed in 70% of patients, moreover, in patients not usually considered amenable to this kind of treatment. Transtympanic steroid treatment is an effective and safe option for patients with sudden sensorineural hearing loss when conventional treatment regimens have failed. Further studies are needed to define the optimal dosage, route of administration and type of steroids. PMID- 18383753 TI - Correlation between vestibulo-ocular reflex and optokinetic afternystagmus in normal subjects and in patients with vestibular system disorders. AB - Optokinetic afternystagmus follows optokinetic nystagmus as an expression of the central velocity storage integrator discharge and its fast phase is beating in the same direction as the previous optokinetic nystagmus. We investigated the correlation between vestibulo-ocular reflex and optokinetic afternystagmus in normal subjects and in patients with bilateral vestibular disorders. The aim of this study was to determine the possible role of optokinetic afternystagmus as a diagnostic test for identifying functional vestibular disorders. The subjects were examined by electronystagmography and vestibulo-ocular reflex, optokinetic nystagmus stare type as well as optokinetic afternystagmus were recorded. They were restrained in a rotatory drum chair, both the chair and the drum could be rotated, independently or coupled. For vestibulo-ocular reflex analysis, we studied post-rotatory-nystagmus from a velocity of 90 degrees s. Optokinetic nystagmus was recorded at a drum velocity of 30 degrees s and the registration continued in total darkness, after the illumination was switched off, to study optokinetic afternystagmus. We considered vestibulo-ocular reflex and optokinetic nystagmus gain, vestibulo-ocular reflex and optokinetic afternystagmus constant of time (tc) defined as the time necessary for the slow phase eye velocity to be reduced to 37% of its initial value. Results demonstrated that vestibulo-ocular reflex gain and ct showed a significant difference only in patients with reduced vestibular reflexia, while optokinetic nystagmus gain was greater only in patients with increased reflexia; optokinetic afternystagmus ct was different from the control group only in patients with hyporeflexia. In conclusion, our results suggest that vestibulo-ocular reflex and optokinetic afternystagmus ct are clinically more useful than the gain alone in testing vestibular disorders with hyporeflexia. On the other hand, we propose a new mathematical and statistical approach to study the temporal evolution of more parameters of the nystagmus. PMID- 18383754 TI - Paroxysmal positional vertigo: the role of age as a prognostic factor. AB - Aim of this study was to examine possible relationships between several clinical aspects of paroxysmal positional vertigo and factors better defined as "intrinsic" to the patient, above all age. The disorder can affect essentially all age groups; nevertheless, the onset of age-linked degenerative processes, such as vascular damage, can have a negative influence--at least in theory--on the pathogenic mechanisms of cupulolithiasis or canalolithiasis. The study was based on the review of 566 patients with the typical form of paroxysmal positional vertigo. Based on age, the patients were divided into two groups, respectively < or =50 years and > 50 years. For the purposes of this study, a series of clinical-laboratory conditions associated with the risk of, or clear, vascular damage were also considered. The results indicate that if there are no clinical or case-history elements that can be attributed to an aetiological hypothesis, the clinical behaviour of paroxysmal positional vertigo is not affected by the age factor. However, the existence of generic vascular damage, hypothesised by the presence of the above-mentioned conditions, influences certain clinical aspects of the disorder, particularly recovery time, the trend of the active phase and the number of relapses. In conclusion, paroxysmal positional vertigo with a presumed vascular aetiology, the incidence of which increases with age, presents a worse prognosis, not only with respect to the "idiopathic" form in childhood but also the "idiopathic" type in the elderly. The lithiasic model responds well to pathogenic interpretation requirements, which envisage macular degeneration with a vascular component. However, the observation, via imaging, of diffuse ischaemic lesions in critical areas of the brainstem and the cerebellum in many "vascular" patients, does not exclude the possibility of alternative pathogenic mechanisms that, in the final analysis, can lead to compromised VOR on a central level. PMID- 18383755 TI - Outcome assessment in patients with chronic obstructive rhinitis CO2 laser treated. AB - Surgical lasers have been used to restore nasal flow in chronic obstructive rhinitis, with a significant improvement in symptoms having been reported in almost all cases. However, evidence supporting the efficacy at long-term, and studies on the assessment of quality of life remain limited. In the present study, efficacy at long term and improvement in the quality of life were assessed in patients with chronic obstructive rhinitis, treated with CO2 laser. A total of 308 patients with chronic obstructive rhinitis were enrolled. The primary outcome measure assessed was the change in score regarding specific and general symptoms, between baseline to 2-4.5 and 7.8 mean years follow-up. Laser turbinotomy restored nasal flow and induced a change in total score which was statistically significant, for specific and general symptoms at the first, second and third follow-up, p < 0.01. CO2 laser turbinate surgery improved symptoms and quality of life in patients with chronic obstructive rhinitis as observed at 2-4.5 and 7.8 mean years follow-up. PMID- 18383756 TI - Otorhinolaryngology-related tuberculosis. AB - Data from personal case histories, from 1984 to 2000 inclusive, are reported in order to contribute to a better understanding of some of the clinical and epidemiological ENT associated TB aspects. Analysis of these data shows that: (1) Like the pulmonary form, ENT localizations are increasing due to the traditional risk factors (immigration, poverty, immunodeficiency, drug addiction). (2) They are generally clinically primitive forms (which are found in extrapulmonary regions as the first expression of tubercular disease) and typically affect young people with a slight prevalence among females. Lymph gland localizations are the most frequent. PMID- 18383757 TI - Myoepithelioma of a minor salivary gland of the cheek: case report. AB - Myoepithelioma is a rare neoplasm of the salivary glands, generally occurring in the parotid gland and less often in the minor accessory salivary gland of the oral cavity. The histological appearance includes solid, myxoid and reticular growth patterns. Vimentin and S-100 protein are very sensitive but non-specific immunohistochemical markers of neoplastic myoepithelium. Conservative surgery is the treatment of choice. A case of myoepithelioma of the minor salivary gland of the cheek with low grade malignancy is described, focusing on clinical behaviour, histopathological and immunohistochemical features and differential diagnosis. PMID- 18383758 TI - Neoplastic transformation of oral lichen: case report and review of the literature. AB - Aim of the present investigation was to analyse the possible malignant transformation of oral lichen planus to carcinoma, especially in the atrophic erosive forms and those displaying plaques involving the top of the tongue. A review has been made of the literature, from 1986 to the present day. This search outlines the relationship between oral lichen planus, hepatitis C virus infection, Epstein-Barr virus infection and the importance of periodic follow-up in all patients with oral lichen planus. The case is described of malignant transformation of oral lichen planus to oral cancer in a female presenting asymptomatic hepatitis C virus infection. The clinical history confirms the most important aspects of the relationship between oral lichen planus and oral cancer. Oral lichen planus should be considered as a precancerous lesion, particularly in patients presenting hepatitis C virus infection, requiring follow-up, at close intervals, starting from 3 months after diagnosis. PMID- 18383759 TI - [Italian Journal of Cardiology. Preface]. PMID- 18383760 TI - [How to use the journal: an ambitious perspective]. PMID- 18383761 TI - [From the Italian Heart Journal, to the Italian Journal of Cardiology, to the Journal of Italian Cardiology]. PMID- 18383762 TI - [Presentation: European practice guidelines for the prevention of cardiovascular diseases in clinical practice]. PMID- 18383764 TI - [Italian application of the European practice guidelines for prevention of cardiovascular diseases]. PMID- 18383765 TI - [Why a painful decision?]. PMID- 18383763 TI - [European practice guidelines on prevention of cardiovascular diseases: executive summary]. PMID- 18383766 TI - [The heart in the city]. PMID- 18383767 TI - [Physiopathology of the left atrium]. PMID- 18383768 TI - [Physiopathology of the left atrium]. PMID- 18383769 TI - [Prehospital fibrinolysis: from the experience of Alta Irpinia to Campania region's network]. PMID- 18383770 TI - [Renin-angiotensin system modulation: instructions for use]. AB - Angiotensin-converting enzyme (ACE) inhibitors and AT1 receptor blockers have long been considered as two classes of drugs with strictly comparable effect in cardiovascular diseases, on the assumption that both classes act on the renin angiotensin-aldosterone system. The results of large clinical intervention trials, which failed to demonstrate any significant difference between the effects of these two pharmacological classes in patients with essential hypertension, acute myocardial infarction and heart failure, supported this concept. The recent observation that a combination of ACE-inhibitors and AT1 receptor blockers improves the prognosis of these pathological conditions better than monotherapy at higher doses focused on the difference between their mechanisms of action. The results of pathophysiological studies have suggested that in the heart, as well as in the kidney, AT1 receptor blockers act in the early stages of the disease, improving left ventricular dysfunction in hypertensive patients and preventing microalbuminuria in diabetic animals. It seems reliable to suggest that AT1 receptor blockers are to be preferred to ACE inhibitors for an early prevention of cardiovascular and renal disease. The new inhibitors of renin activity may further amplify our chances, also blocking the negative effects mediated by angiotensin II escape and by stimulation of the prorenin/renin receptors. PMID- 18383771 TI - [Air pollution exposure as an emerging risk factor for cardiovascular disease: a literature review]. AB - Several epidemiological studies have demonstrated a consistent statistical association between cardiovascular disease and air pollution exposure. In this review we describe the nature of the most common ambient-air pollutants (either gaseous or particulate of different size); then, we examine the pathophysiological pathways linking the air pollutants with their cardiovascular effects (arterial vasoconstriction, systemic inflammatory response, enhanced thrombosis, a propensity for arrhythmia). A comprehensive review of the literature on air pollution and the rate of hospital admission, morbidity, and mortality due to cardiovascular reasons are also provided. In the last section, we review the most recent studies that have outlined the correlation between the onset of acute myocardial infarction and the level of outdoor air pollution. In conclusion, scientific evidence is growing in support of the hypothesis that ambient air pollution acts as a risk factor for cardiovascular disease, and may trigger the onset of acute myocardial infarction. PMID- 18383772 TI - [Transcatheter cryoablation in children: state of the art]. AB - A new transcatheter ablation method has recently made its appearance in leading electrophysiology laboratories: cryoablation. In contrast with radiofrequency, this method produces a permanent lesion due to cell necrosis, caused by application of very low temperature chilling to the tip of special ablation catheters placed against the area of the heart causing arrhythmia. The benefit of this system over radiofrequency ablation is its ability to find the most suitable site for ablation through transitory electrical paralysis of the heart tissue in contact with the catheter tip, chilled to -30 degrees C (cryomapping). If the site is suitable, the tissue causing the arrhythmia loses its excitability. Moreover, the stability of the catheter tip, which adheres to the myocardium, enables arrhythmia inducibility through programmed atrial stimulation to be assessed without dislodging the catheter. A permanent lesion is created only subsequently, with further chilling to even lower temperatures (cryoablation). In the last 3 years the success rate of cryoablation in pediatric patients has increased and long-term complications have not been described yet. We therefore consider that cryoablation should be considered the treatment of choice for atrioventricular nodal reentrant tachycardia and parahissian, anteroseptal and right midseptal accessory pathway atrioventricular reentrant tachycardia in school-age children. In fact cryoablation is thus "made to measure" for pediatric patients because if an atrioventricular block is observed, stopping the procedure is quickly followed by the restoration of normal atrioventricular conduction. PMID- 18383773 TI - [Medical-legal evaluation of Brugada syndrome]. AB - Brugada syndrome is a congenital arrhythmogenic disease, characterized by alterations in sodium channels in the absence of structural myocardial changes. It leads to ventricular electrical instability, causing arrhythmias with high risk of sudden cardiac death. Patients affected by Brugada syndrome show electrocardiographic alterations either at baseline or after stimulation. Symptomatology and electrocardiographic alterations are the most important prognostic elements. The only effective treatment is the implantation of an automatic implantable cardioverter-defibrillator, which dramatically reduces arrhythmic mortality. This device is used only in symptomatic patients for abortive sudden cardiac death or syncope with electrocardiographic alterations, whereas in most patients just a careful follow-up is required. In this article we evaluate the medical-legal aspects of patients with Brugada syndrome, in forensic pathology, social insurance, private insurance, medical professional liability and ability to work. First of all, we analyze the question of medical professional liability, mostly in case of a wrong identification of prognostic factors. Then we consider the medical-legal evaluation of Brugada syndrome in the field of social insurance, in relation to the clinical picture and particularly to the frequency of arrhythmic events. As far as private insurance is concerned, we dwell upon patients' insurability in the various types of policies, refundable medical expenses and evaluation of pathology in health insurance. Finally, we discuss the question of ability to work, with special regard to armed forces and police, whose committees do not provide a specific evaluation for Brugada syndrome by making identical use of automatic implantable cardioverter defibrillators and pacemakers, thus ignoring the great clinical differences between these two devices. PMID- 18383774 TI - [Implementation of an emergency clinical pathway for ST-elevation myocardial infarction in the Lazio Region: results of a pilot study]. AB - BACKGROUND: Timely reperfusion therapies (primary angioplasty and pre-hospital thrombolysis) remain a key component in improving the survival of patients with ST-segment elevation myocardial infarction (STEMI). The Lazio Region emergency organization has a complex mixed logistic (the large city of Rome, presence of complex orography), therefore the use of telemedicine technologies by the emergency medical system (EMS) is mandatory. Emergency clinical pathways (ECP) for the management of STEMI patients were designed, focusing on early pre hospital diagnosis and best appropriate treatment through the ECG transmission and teleconsultation among EMS and cardiologists in coronary care units (CCU). METHODS: To evaluate the effectiveness of ECP-STEMI in the current practice, a prospective observational cohort study of ambulance-transported patients with cardiovascular symptoms was conducted in a selected area of the Lazio Region during a 6-month period. The implementation of the ECP was carried out by educational activities for the EMS personnel based on the "experiential learning" methods. RESULTS: From October 2005 to March 2006, 287 patients were enrolled in the study and a pre-hospital ECG was performed in 66% of them. One hundred and fifty-two patients were referred to hospital and only 34 had discharged diagnosis of acute myocardial infarction, of whom 23 were STEMI. In the 34 acute myocardial infarction patients the medium time from "call to the EMS" to "arrival to the hospital" was 41 min (range 29-63 min) and 3 had their ECG telematically transmitted from the ambulance to the CCU. All of these cases were STEMI. Twenty eight acute myocardial infarctions were discharged alive, 2 were transferred in other hospitals, 4 died. No patients received pre-hospital thrombolysis. Prior to the ECP implementation the ECG for STEMI patients has never been transmitted by EMS to the CCU in the Lazio Region. CONCLUSIONS: Our study suggests that adherence to ECP improved the appropriateness of STEMI patient referral and treatment in the CCU in the Lazio Region. The EMS personnel, during the study, showed a high interest in the protocol trying to change their current practice. The Regional Administration plans to expand the utilization of ECP to all regional emergency network (EMS and Emergency Departments) and to improve its use. PMID- 18383775 TI - [Manual thrombus aspiration with the Export catheter during primary and rescue angioplasty. Predictors of failed myocardial reperfusion]. AB - BACKGROUND: Manual thrombus aspiration seems to improve myocardial reperfusion after coronary angioplasty in patients with ST-elevation acute coronary syndrome. We sought to assess the independent variables of complete myocardial reperfusion after primary and rescue coronary angioplasty with use of the Export manual thrombus aspiration catheter. METHODS: Myocardial reperfusion were judged complete if ST-segment resolution were >70% at 60 min post-procedure ECG. Sixty three consecutive patients undergoing primary (81%) or rescue angioplasty (19%) with use of the Export catheter were analyzed. RESULTS: Forty-three (68%) patients had a complete myocardial reperfusion and 20 (32%) patients did not. The independent predictors of failed myocardial reperfusion, at multivariate analysis, were: heart rate at hospital admission (odds ratio [OR] 1.05; 95% confidence interval [CI] 1.01-1.09; p = 0.007), time to angioplasty >3 h (OR 6.07; 95% CI 1.31-27.97; p = 0.005), left anterior descending coronary artery involvement (OR 8.8; 95% CI 2.12-36.4; p = 0.003). CONCLUSIONS: The present study shows that in the setting of primary or rescue angioplasty with use of Export manual thrombus aspiration catheter, patients with high heart rate on admission, an ischemic time >3 h and left anterior descending coronary artery involvement are at higher risk of failed myocardial reperfusion. PMID- 18383776 TI - [Tako-tsubo cardiomyopathy complicated by left apical thrombus]. AB - Tako-tsubo cardiomyopathy is a rare syndrome usually described in postmenopausal women, with a presentation mimicking an acute myocardial infarction. We report an unusual case of tako-tsubo cardiomyopathy complicated by left ventricular apical thrombus. PMID- 18383777 TI - [Acute profound thrombocytopenia associated with tirofiban: clinical approach to diagnosis and therapeutic management]. AB - Acute profound thrombocytopenia is a serious complication of glycoprotein IIb/IIIa inhibitor therapy characterized by a precipitous decline in platelet count to <20 x 10(9)/l within few hours of therapy initiation. This potentially devastating event is extremely uncommon in patients receiving small molecule inhibitors such as eptifibatide and tirofiban. The authors present a case of a patient with acute coronary syndrome scheduled for urgent percutaneous coronary intervention who developed acute profound thrombocytopenia during treatment with intravenous tirofiban. The platelet count decreased to a nadir of 3 x 10(9)/l 2 h after initiation of inhibitor administration. All other possible etiologies of thrombocytopenia were unlikely and the low platelet count had to be attributed to tirofiban. Tirofiban was discontinued and the patient was treated with steroids, immunoglobulin and platelet transfusion. No bleeding events occurred and the patient was discharged on day 18. Interestingly, the patient underwent a percutaneous coronary intervention 1 month before and received the same dosage regimen of tirofiban without complications. Implications for the incidence, diagnosis and treatment of thrombocytopenia disorders are discussed. PMID- 18383778 TI - [Anomalous origin of the right coronary artery from the left anterior descending coronary artery]. PMID- 18383779 TI - Intestinal parasites in school aged children and the first case report on amoebiasis in urinary bladder in Tripoli, Libya. AB - A total of 50 fresh fecal samples were collected from children of either sex and examined by direct wet films and Telemann's concentration technique. The results were based on a single stool per person study of the total children ranging from 5-18 years of age, 8 different types of intestinal parasites were recorded. The incidence of infection with one or the other parasites was found to be 42%. Double or mixed infection was not uncommon. The infection rates of helminthes were Hymenolepis nana 6%, Taenia saginata 2%, Ascaris lumbricoids 20%, Trichuris trichura 14% and Enterobius vermicularis 4%. The rates of protozoa were Entamoeba histolytica 4%, Entamoeba coli 4% and Giardia lamblia 2%. The problem was created by parasitosis in endangering children's health are more increasing in Tripoli District. PMID- 18383780 TI - Effect of aflatoxin on the coccidial infection in broilers. AB - Two hundred of one day old chickens were divided into four equal groups and kept for the end of experiment. The first group was kept as control negative, the second group received 1 ppm of dietary aflatoxin from day zero of chick life till the end of present study, while the third group was given 4x104 sporulated oocysts of Eimeria sp., the group four were obtained coccidial oocysts and aflatoxin in their rations. The combination of aflatoxin and coccidia, produced higher mortality rate, higher faecal scores and increased oocysts output than those chicks received aflatoxin or coccidia only. Body weighs and efficiency of feed utilization were decreased in all treated groups. The maximal losses of body weight and efficiency of feed utilization were noticed in chicks infected with Eimeria sp. and at received aflatoxin in their ration. The levels of total serum proteins, gamma globulins, calcium and phosphorus were decreased in chicks infected with coccidia and received dietary aflatoxin. Total bilirubin and SGOT activity were higher in chicks infected with Eimeria sp, and obtained aflatoxin. PMID- 18383781 TI - Prevalence of echinococcosis in street dogs in Tripole District, Libya. AB - Cystic echinococcosis due to E. granulosus is a serious public health and livestock economy problem in Libya. Kato thick smear examination of 50 street dogs stools showed that they had Echinococcus granulosus (58%), Taenia spp. (14%), Diplydium caninum (16%), Toxocara canis (121%) and 20% were parasite-free. The stool examination using Kato thick smear was more sensitive and more specific that the indirect haemaggutination test. The results were discussed with general review of the disease in Libya. PMID- 18383782 TI - Evaluation of the in vitro effect of Nigella sativa aqueous extract on Blastocystis hominis isolates. AB - The effect of Nigella sativa aqueous extract was evaluated against the in vitro growth of 2 different isolates of the intestinal protozoan parasite Blastocystis hominis. Different concentrations (10, 100, 500 microg/ml) of Nigella aqueous extract and metronidazole, an active standard drug for B. hominis, were incubated with B. hominis isolates in culture media at 37 degrees C. Their possible effect on B. hominnis living cell count (LCC) was assessed on Day 1, 3 & 6. The aqueous extract of N. sativa at concentrations of 100 and 500 microg/ml showed a potent lethal effect on both B. homninis isolates, but with different extent. There is no significant difference between the inhibitory effect of N. sativa and metronidazole on the LCC on the 6th day. On assessment of living cell rate (LCR) which calculate percentage rate of living cell, N. sativa at 500 microg/ml concentration has a significant inhibitory effect on both isolates. So, it is considered as the most active concentration of Nigella aqueous extract. These results prove that N. sativa aqueous extract could be useful in the treatment of B. hominis. PMID- 18383783 TI - Host-induced phenotypic differences in Egyptian Heterophyes heterophyes (Digenea: Heterophyidae). AB - A comparative morphometric study identified host-related variations in Heterophyes heterophyes. In one study, variations in adult H. heterophyes obtained from different definitive hosts were examined. Significant differences were in size and dimensions of body organs of adults collected from dogs, cats and albino-rats experimentally infected with encysted metacercariae (EMC) from mullet (Mugil cephalus). Hypotheses on the sources of host-dependent variations were interpreted in terms of adaptation to conditions created by the host. In a second study, comparative morphometric analysis of adults recovered from puppies experimentally fed on EMC isolated from five sympatric fish species was done. No significant differences were in adults H. heterophyes recovered from puppies feed on EMC of different fishes. Constancy in morphological characters of adults reared in the same host, independent of a change of fish was seen. So, the variations depend upon host species where worms develop, and host selection may determine the worm morphology and phenotypic plasticity. PMID- 18383784 TI - Ectoparasites of commensal rodents in Talkha Center, Dakahlia Governorate, Egypt. AB - The ecto-parasites infesting commensally rodents were collected from the different localities in Talkha Center (Dakahlia Governorate) from April 2006 to March 2007. The seasonal abundance of rodent number and rat index was 52 (0.58) in spring, 27 (0.3) in summer, 39 (0.22) in autumn and 17 (0.05) in winter. From 135 rodent species and rat index was Rattus norvegicus N=33 (0.24), R. r. frugivorous N=39 (0.29); R. r. alexandrinus N=48 (0.36) and Mus musculus N=15 (0.11). From 388 ecto-parasite infested rodent collected number and ecto index was fleas N= 114 (0.84 flea/rat), Lice N=93 (0.69 lice/rat), Mites N = 165 (1.2 mite/rat) and larva of ticks N=16 (0.12 tick/rat). No doubt, rodents and their ectoparasites played a serious role in spreading and transmission of zoonotic diseases to human and animal. PMID- 18383785 TI - Assessement of peritoneal adhesions due to starch granules of surgical glove powder an experimental study. AB - Introduction of foreign material into abdominal cavity during surgery causes irritation of peritoneum leading to postoperative (PO) adhesions. To assess and prevent PO adhesions, an experimental study was done to assess the caused by starch granules of the surgical glove powder and to avoid complications using the effect of normal saline and low molecular weight heparin (LMWH). Thirty Wister Albino male rats were subjected to laparotomy and ileocaecal abrasions were done mechanically to induce postoperative peritoneal adhesions. Rats were divided into 3 groups, 10 animals each. The sterile starch powdered gloves were used in the laparotomy procedure. The basic procedure performed in the three groups was serosal abrasions at the ileocaecal region and using the 2-layer technique in the closure of the abdominal cavity. In GI no further management was performed than the basic procedure. In GII & GIII before closure normal saline and (LMWH) was spilled into the peritoneal cavity respectively. The results showed that in GI, there were severe firm intraperitoneal adhesions especially at the site of the ileocaecal abrasions. This was proved macroscopically and histologically. The reactions detected in biopsies of GII & GIII were lesser in intensity compared to GI. Morphometric assessment of collagen deposition in the intra-peritoneal adhesions revealed a significant decrease in GII & GIII compared to GI. PMID- 18383786 TI - Zoonotic cutaneous leishmaniasis: reservoir host and insect vector in north Sinai, Egypt. AB - Cutaneous leishmaniasis (ZCL) is a protozoan disease well documented not only in Egypt, but in nearly all the East Mediterranean Countries. Sinai Peninsula was a sparsely populated area where sporadic cases of ZCL were reported with the reconstruction of Sinai and people coming in and out, it was indicated to study the status of ZCL in North Sinai Governorate, the reservoir host(s) and insect vector(s) in Sinai. In the present study, the six species of rodents were trapped from areas or nearby areas where human ZCL cases were detected. Rodents (50) were Mus musculus (10), Rattus r. alexandrinus (18), R. norvegicus (2), Gerbillus gerbillus (4), G. pyramidum (12) and Jaculus jaculus (4). The rodents were examined clinically for any skin lesion or even nodule, particularly in head and tail. One G. pyramidumn had natural infection with L. major as indicated by smears and culture, but typing was not done. The spot light surveys for Phlebotomus were carried out by the sticky paper traps and the CDC light traps in four main centers; Al Hassanah, Nakhil, Al Arish, and Bir Al-Abd. A total of 1320 sandflies were identified. They were P. papatasi (1150) and P. sergenti (170) in a ratio of 7:1. A total of three isolates of zymodeme London 70 undistinguished from the formerly obtained human and rodent isolates were enzymatically identified in P. papatasi. PMID- 18383787 TI - A new host and locality record: Gnathia sp. (Isopoda: Gnathiidae) on the barred mudskipper, Periophthalmus argentilineatus Valenciennes, 1837 (Perciformes: Gobiidae) from Tanzania. PMID- 18383788 TI - Effect of warfarin anticoagulant rodenticide on the blood cell counts of Rattus norvegicus and Rattus rattus. AB - The effect of the warfarin LD50 on the counts of the blood cells of Norway rat, Rattus norvegicus and roof rat, Rattus rattus from Giza and Qualyobia Governorates was investigated in the laboratory. Warfarin toxication caused significantly different blood cell counts in treated rats compared to control ones. Warfarin significantly decreased the total erythrocytic count and increased the total leucocytic count of treated rats. But, it caused significant de-crease in neutrophrils, eosinophils, basophils and monocytes, but significant increase in lymphocytes. The data also showed that in survived rats, WBC count equaled the normal level or slightly exceeded it at 33/43 days post treatment. PMID- 18383789 TI - Mirazid in treatment of human hymenolepiasis. AB - The efficacy and safety of Mirazid in treatment of human hymenolepiosis were carried out in a rural village in Talkha Center (Dakahlia G.). Kato thick smear stool examination showed 51 cases of Hymenolepis nana (9 of them had concomitant parasitosis), two cases of H. dimninuta. Mirazid was given in a dose of 10 mg/kg/d for nine consecutive days an hour before breakfast for hymenolepiosis cases, as capsule for adults and suppository for children. All cases were subjected to history taking before treatment and six weeks post-treatment and stool examination was repeated as well weekly for six weeks post-treatment. There was overt clinical improvement. Side effects were negligible. Parasitologic cure rate was 40/41 or 95.2% for H. nana one week post-treatment, and 100% for H. diminuta one week post-treatment, up to six weeks follow-up for all treated patients. Two unresponded H. nana patients were cured by another Mirazid course. PMID- 18383790 TI - Intestinal permeability in Hymenolepis nana as reflected by non invasive lactulose/mannitol dual permeability test and its impaction on nutritional parameters of patients. AB - Assessment of Hymenolepis nana infection among 102 children and adults of both sexes (5-16 years) residing 2 Welfare Institutes (Giza and Cairo) showed a prevalence of 22.33%. The effect of H. nana on intestinal permeability and on nutritional parameters of patients was studied. A total of 46 subjects were divided into 2 groups: GI (20 H. nana patients) and GII (26 parasite-free control). Both groups were subjected to lactulose/mannitol dual permeability test, anthropometric study, estimation of vitamin B12 and folate levels in plasma and estimation of haemoglobin (HB)%, RBCs and WBCs counts and haematocrite value (HCT%) for anaemia. The H. nana patients showed significant higher percent (P = 0.04) of altered intestinal permeability versus controls denoting intestinal leakage, significant means lower levels of vitamin B12 (P = 0.01) and folate (P < 0.04) in blood plasma versus control denoting liability to vitamin B12 & folate deficiencies. Means value of HB%, RBC & WBC counts and HCT% showed generalized decrease but without significant difference in H. nana patients and control denoting anaemia liability. The percent of stunting (HAZ < or =2) and of wasting (WAZ < or =2) were higher among H. nana patients versus controls but without significant difference (P = 0.19 & P = 0.47 respectively). PMID- 18383791 TI - Susceptibility of Culex pipiens complex to some insecticides in Qatar. AB - Insecticides susceptibility tests were conducted on Culex pipiens larvae from both Doha and Al-Rayan. The insecticides used were chloropyrifos, propetamphos (Organophosphorous), cyphenothrin, cyfluthrin (Synthetic) and etofenprox. Larvae in Al-Rayan district were more susceptible to all insecticides than in Doha. The chlorophyrifos and cyfluthrin were the most effective larvicides, cyphenothrin was moderate and propetamphos and etofanprox were the least ones. PMID- 18383792 TI - Touchdown PCR, ELISA and stool examination for early diagnosing of Schistosoma mansoni in mice. AB - A detection assay for Schistosoma mansoni DNA in mouse serum samples based on touchdown PCR was evaluated. Fifty four mice (7 weeks old) were infected with 60 +/- 10 cercariae of S. mansoni (Egyptian strain) through tail skin. Three days and 1-8 weeks post-infection (PI), six mice were randomly chosen from the infected ones and blood was collected by cardiac puncture. Sera and faeces were collected for 8 weeks PI. Antisoluble egg antigen (SEA) IgG for S. mansoni was detected in infected mice sera of at 3 weeks PI by ELISA. The earliest deposition of eggs in the small intestines was at the 5th week PI. The eggs were detected in faeces at the 8th weeks by modified Kato's thick-smear. However, S. mansoni DNA was detected by touchdown PCR in sera at 2 weeks PI. The intensity of S. mansoni DNA bands in infected mice sera detected in polyacrelamide gel electrophoresis increased gradually during the experimental period. Data recommended touchdown PCR potential tool for the early diagnosis of S. mansoni infection. PMID- 18383793 TI - Efficacy of medical ozone in attenuation of murine Schistosomiasis mansoni infection morbidity. AB - Mice were divided into 3 groups: non-infected control, S. mansoni infected non treated for 10 weeks and group receiving ozone intraperitoneally for 21 days 10 weeks postinfection. Results showed that ozone therapy in chronic schistosomiasis mansoni decreased worm burden, increased number of dead eggs, decreased both mature and immature eggs, increased RBC count, improved hemoglobin concentration with reduced reticulocytes %, as well as increased lymphocyte %, decreased neutrophil % and markedly increased eosinophil %. Platelet count was increased and bleeding time was markedly shortened, markers for hepatic function were improved, and malondialdehyde concentration decreased in liver and spleen but increased in intestine. Reduction of associated hepato-splenomegaly was noticed, with reduction in number and size of hepatic and intestinal granulomas, collagen area % and number of macrophages, with preservation of hepatic and intestinal histological profile in almost all areas. The results point to medical ozone as a promising agent to complement schistosomiasis mansoni specific treatment, helping to attenuate infection morbidity. PMID- 18383794 TI - Immunochemotherapy for cryptosporidiosis in immunosuppressed mouse model. AB - Immunochemotherapy as a dual regimen (Nitazoxanide NTZ and Interferon gamma INF gamma) and a triple one (NTZ, INF-gamma & Paromomycin PRM), administered to immunosuppressed Cryptosporidium infected mice for 10 days (4th-13th day post infection) was evaluated during and after treatment by determination of parasite count in ileum, associated histopathological changes, oocyst count in Kinyoun's acid fast stained faecal smears, percent reduction in oocyst excretion and cure rate. Both regimens induced nearby efficacy (P > 0.05) with significant reduction in parasite count in the ileum on 7th (P < 0.01) & 14th (P < 0.001) P.I. days, partial regression of histopathological changes and reduction in oocyst count from the 2nd day post-treatment. Oocyst excretion reduction percent was reached zenith on 13th P.I day in both dual (95.76%) and triple (94.86%) regimens (P > 0.05). Complete cure was not achieved. Three days post-treatment relapse occurred in both regimens (P < 0.001) increase in oocyst count (P < 0.01) increase in parasite count in ileum, more severe histopathological changes with rapid deterioration and then, death of all remaining treated mice PMID- 18383795 TI - Efficacy of garlic extract on hepatic coccidiosis in infected rabbits (Oryctolagus cuniculus): histological and biochemical studies. AB - The rabbits were divided into three groups, of 12 rabbits each. G1 was the (non infected non-treated) as control, G2 was the (infected-non treated), and G3 was the (infected and treated) rabbits. Each rabbit in the infected groups were given (10(3)) sporuleted oocysts of Eimeria stiedae per rabbit after forty five days exactly. Faecal sample of rabbits from each group were examined each day post infection till oocysts appeared in faeces. The treatment was given by using suitable dosage of garlic according to body weight. After 15, 21, 28, & 35 days post-treatment faecal oocysts were output. Biochemical parameters as serum liver function (ALT, AST, GGT & ALP) that denoted the he-patic cells injury. The results showed a significant differences in the mean values of oocysts shedding and their mean number in bile ducts between Gs 2 &3 from the 15th day post infection (PI) (mean +/-SD:40.33 +/- 16.72 & 25.17 +/- .56 respectively) till the experimental end on the 35th day (55.75 +/- 19.79 & 0.94 +/- 1.43 respectively). The histopathological alterations were in liver of G2 at the experimental end. Coccidiosis in G2 induced histopathological alterations in liver tissue, marked cytoplasmic vacuolations in hepatocytes with clear signs of karyolysis, and dilatation of sinusoids with increase in Kupffer cells. Leukocytic infiltration around congested blood vessels was noticed. Efficacy of garlic on E. stiedae in infected Gs was resident. The liver of G3 regained almost normal appearance compared to control. PMID- 18383796 TI - Malaria in Egypt, Saudi Arabia and Yemen: a clinical pilot study. AB - Imported malaria is a health problem and needs continuous monitoring as many clinicians are not aware of it. In Yemen malaria is the main public health problem. Malaria cases were 16 in Almaza Military Fever Hospital, Cairo, 53 in Saudi Hospital at Pilgrimage, Yemen and in Saber Hospital at Aden, Yemen were studied. 9 cases (56.2%) of P. falciparum in Cairo were imported and 7 cases (43.8%) acquired P. vivax locally (October 2003 to July 2004). They were all treated successfully by chloroquine. An imported case (6.3%) died by cerebral malaria due to delayed diagnosis. Five imported cases (31.3%) had severe malaria. In Pilgrimage, an infant (1.9%) had congenital malaria, 17 cases (32.1%) had severe malaria and 2 (3.8%) died by cerebral malaria. 43 patients (81.1%) had P. falciparum and 10 patients (18.9%) had P. vivax. All patients were treated by parenteral or oral quinine. In Aden, one patient (5%) suffered diarrhea without fever, early blood film was negative, and was positive later on. 18 cases (90%) had P. falciparum, 2 (10%) had P. vivax. 4 cases (20%) had severe malaria & a patient (5%) died by cerebral malaria. Patients in Aden severe cases were successfully treated by intramuscular artemether followed by oral Fansidar, and mild ones were treated by oral Quartem. PMID- 18383797 TI - Antibody isotypes in urethral swabs of symptomatic and asymptomatic men infected with Trichomonas vaginalis. AB - Trichomoniasis may be asymptomatic or symptomatic in both sexes. The outcome of infection depends on the virulence factors of T. vaginalis, but these factors remain unclear. Genetic variability of the isolates and the host's immune response are likely to be key factors in that respect. Symptomatic and asymptomatic males infected with T. vaginalis were compared regarding the differences in antibody subclasses response in the urethral samples. In symptomatic cases there was a significant elevation in IgM, IgG1 & IgG2b levels in urethral samples, and a little, non-significant rise in IgG2a levels. However, there were no statistically significant differences between levels of IgA, IgG3 & IgG4. The results showed that specific IgG1 & IgM and to a lesser extent IgG2 may be involved in established symptomatic trichomoniasis in men, compared to asymptomatic ones. PMID- 18383798 TI - Ultrastructure characteristics of Besnoitia darlingitachyzoites Brumpt, 1913 (Protozoa: Sarcocystidae) of the Michigan strain MIBD1. AB - Tachyzoites of Besnoitia darlingi Brumpt, 1913 were redescribed based on new materials isolated from Virginia opossums (Didelphis virginiana, Kerr) from Michigan, U.S.A. Tachyzoites of the MIBD1 strain were propagated in bovine turbinate cell culture for more than two years. A comparison with previously described tachyzoites of the B. darlingi OP1 strain from Mississippi, USA revealed some morphological differences despite the remarkable genetic homogeneity between the two B. darlingi strains. MIBD1 tachyzoites were distinguished from OP1 tachyzoites by having more rhoptries, and fewer and haphazardly distributed micronemes at the conoidal end. This morphological heterogeneity between tachyzoites of the two strains suggests the role of geographical isolation in the Michigan strain. New morphological features of B. darlingi tachyzoites were described. PMID- 18383799 TI - Natural infection of fresh and brackish water fish with heterophyid encysted metacercariae in Dakahlia Governorate, Egypt. AB - The encysted metacercariae (EMC) of heterophyids in the fish species; Mugil caphalus, M. capito, M. auratus, Oreochromis n. niloticus (=Tilapia nilotica), and Tilapia zillii trapped brackish and fresh water fishes were isolated by the artificial digestion method and identified. The EMC by morphometric analysis proved to be seven phenotypical distinct types. Cross-matched seven groups of clean laboratory bred puppies each was experimentally infected with a distinct EMC type. The recovered adult flukes were compatible to seven species of six genera of family Heterophyidae. These were Heterophyes heterophyes, H. aequalis, Pygidiopsis genata, Haplorchis yokogawai, Prohemostomumn vivax, Phagicola sp. and Stictodora sp. The EMC of the seven heterophyids were recovered from all the brackish water fishes, M. caphalus, M. capito, M. auratus, O. niloticus and T. zillii. On the other hand, the EMC of P. genata, Haplorchis yokogawai and Phagicola sp. were recovered only from the fresh water fish, O. niloticus and T. zillii. All the EMC was re-described and photographed. PMID- 18383800 TI - Intestinal parasitic infections among school children in Tripoli, Libya. AB - An epidemiological survey was carried out in Tripoli district to collect baseline data on the prevalence of intestinal parasites. A sample of 486 school children aged from 5 to 14 years old provided morning stool samples as part of a pilot study of a larger national project. Questionnaires were distributed and general information on each child was recorded. Posters and leaflets were used extensively to increase the public health information and awareness about parasitosis. The results revealed that 14.6% of children were infected with at least one intestinal parasite. Double infections were in 2% of them. Giardia lamblia was the common parasite, followed by Enterobius vermicularis and then Ascqris lumbricoides. This was a preliminary approach to clarify the status on intestinal parasites since reliable reportable data being still comparatively lacking, which might be a guide for a feasible national control program. PMID- 18383801 TI - Antigen capture immuno-chromatographic strip format in detecting parasite specific lactate dehydrogenase to diagnose malaria in nonimmune patients. AB - Several rapid diagnostic test devices (RDT) based on detection of malaria antigen in the whole blood were developed. OptiMal test the presence of parasite-specific lactate dehydrogenase (LDH) enzyme using three monoclonal antibodies was used. Two monoclonal antibodies were pan-specific and recognized all malaria species. The third one was specific only for Plasmodium falciparum. The parasite antigens were detected using an antigen-capture immunochromatographic strip format. One hundred-nine malaria positive and 730 malaria negative cases diagnosed by microscopy were included. 75/109 were P. falciparum 26 as P. vivax, 3 P. malariae and 5 mixed infection of P. falciparum & P. vivax. The RDT showed a low sensitivity (85%, 95% Confidence Interval [CI], 79-92%) with a much lower sensitivity in detecting species other than P. falciparum as well as in mixed infections. The sensitivity was 50% for less than 200 parasites/micro. The sensitivity of OptiMal for P. falciparum was 87% (95% CI, 79-94), 81% (95% CI, 66 96) for P. vivax, and failed with P. malariae. Mixed infections were misdiagnosed as Pfalciparum. The sensitivity of OptiMal was quite good in detecting both P. falciparum & P. vivax (98%; 95% CI, 97-99 & 100%; 95% CI, 100-100 respectively) and 99% (95% CI, 98-99) for all species. The positive and negative ratio for all malaria species was: (+LR = 62.3, -LR = 0.01); for P. falciparum (+LR = 38.9, -LR = 0.01) and for P. vivax (+LR = 0.8077/0, -LR = 0.2). The test value to assess drug resistance in post treatment days was discussed. PMID- 18383802 TI - Contrast staining of Trichinella spiralis larvae in fresh, frozen and formalin preserved muscles. AB - Fresh, frozen-thawed and formalin-preserved muscle samples heavily infected with Trichinella larvae were cut into several pieces and stained with Giemsa and Leishman and the reference Haematoxylineosin (H & E) stain. Observation under microscope revealed that both muscle larvae and nurse cells in fresh and formalin preserved specimens appeared as purplish blue structures contrasting with the pinkish color of non-infected muscle fibers in both Giemsa and Leishman stains. These findings were confirmed in H & E stained samples. However, frozen samples did not show contrast stain. PMID- 18383803 TI - Immunolocalization of Schistosoma mansoni and Schistosoma haematobium antigens reacting with their Egyptian snail vectors. AB - The reaction of the haemolymph and the tissue of infected intermediate hosts, Biomphalaria alexandrina and Bulinus truncatus to Schistosoma mansoni and S. haematobium antigens were investigated using the indirect immunoperoxidase technique. A new technique, Agarose cell block was used in collection of haemolymph which helped in collecting plenty of well formed cells in comparison to the ordinary one using the cytospin. Collected haemolymph and prepared tissues of uninfected and infected B. alexandria and B. truncatus were fixed and then reacted with anti-S. mansoni and anti-S. haematobium IgG polyclonal antibodies. The haemolymph and tissue of infected B. alexandrina and B. truncatus gave a positive peroxidase reaction represented by a brown colour. In haemolymph, the positive peroxidase reaction was detected mainly in the cytoplasm of the amoebocytes. In the tissue, it was detected in epithelial cells lining the tubules, male cells in the lumen of the tubules and in female oogonia cells along the periphery of the tubules. The similarity in the strength and distribution of positive reaction in B. alexandrina and B. truncates was observed as compared to control. Thus, the immunoperoxidase technique proved to be an effective indicator for the schistosome-antigen in the snails. PMID- 18383804 TI - Long term formalin preserved stool specimens for detection of intestinal parasites from school aged children in Tripoli, Libya. AB - A total of 949 single stool samples were collected from school aged children (5 14 years old) in the city of Tripoli. The samples were preserved in 10% formalin and examined by routine microscopy using normal saline and Lugol's iodine preparations as well as the formol ethyl concentration method after a storage period of twelve months at room temperature. Of 949 samples examined 4.5% were positive. Giardia lamblia and Entamoeba coli were the only protozoan parasites identified with an infection rate of 3.2% & 1.3% respecttively. No helminthes were detected in any of the samples. About 14% of the children had intestinal yeast infection Candida albicans in their stool of which 0.63% was infected with intestinal parasites. No distortion or alteration of morphology was observed particularly in G. lamblia. Preservation in 10% formalin is a very productive means for the accurate identification of protozoan parasites. PMID- 18383805 TI - Studies on the juveniles of a species of Anisakis (Nematoda: Anisakidae) from the orangespotted trevally, Carangoides bayad (Carangidae), from the Red Sea, Egypt. AB - One out of 45 (2.2%) Orange-spotted trevally, Carangoides bayad (Carangidae), collected off the coast of Egypt in the Red Sea was naturally infected with juveniles of a species of Anisakis (Anisakidae). Most of the juveniles were found free in the body cavity and unencapsulated on the surface of the liver. The morphology of juveniles was studied using both light microscopy and SEM. The anterior extremity of larvae had a circle of 4 papillae; the characteristic boring tooth, and lateral amphids arranged around a triangular mouth; a cylindrical, esophagus that is light colored with muscular and ven-tricular portions; colorless excretory canals; excretory pore situated just behind the boring tooth. The cuticular surface of the entire body except the cephalic region was striated with discontinuous, undulating longitudinal bands and transverse striations. The posterior extremity was rounded with a distinct mucron. The external morphological features of the juveniles were compared to the previous described genera and species in family Anisakidae harboring the Red Sea fishes. PMID- 18383806 TI - Flowcytometric analysis of T-lymphocytes and serum tumour necrosis factor alpha (TNF-alpha) levels in schistosoma mansoni patients. AB - Analysis of T-lymphocytes by flowcytometry, estimation of serum TNFa level by solid phase enzyme amplified sensitivity immunoassay (EASIA) and IHAT were done for chronic schistosomiasis mansoni patients without hepatic fibrosis, with hepatosplenomegaly and 20 healthy controls. The sensitivity and specificity of IHAT in schistosomiasis mansoni were 85% & 90% respectively. Chronic schistosomiasis mansoni patients showed increase in CD8% (27.3 +/- 5.3) and decrease in CD4% (44.2 +/- 4.68). Hepatosplenomegaly cases showed increase in CD4% (46.5 +/- 4.1) & decrease in CD8% (23.2 +/- 2.18). Serum level of TNF-a was significantly higher in cases with hepatosplenomegaly compared to either cases of chronic schistosomiasis mansoni or controls. No significant difference was between chronic schistosomiasis mansoni patients and controls. A correlation between hepatosplenonomegaly and increase of CD4 and/or decrease of CD8 and significant high level of TNF-a indicated TNF-alfa role in granuloma formation. PMID- 18383807 TI - Risk factors contributing to fascioliasis endemicity in a focus in Dakahlia Governorate 1-human host. AB - Zoonotic fascioliasis is a problem not only in Dakahlia Governorate, but also in other Egyptian Governorates. Two hundreds & twenty patients randomly selected with suggesting manifestations were examined for fascioliasis. A total of 23 (10.4%) were positive by Kato thick smears, of which 21 were from Kafr El-Hessah and two from Oweish El-Hager. The proven human fascioliasis was examined for anti Fasciola antibodies by IHAT & ELISA (Fhes), haemoglobin level, eosinophils percent, serum bilirubin & liver function tests. IHAT gave 82.61% positive results (19/23), and ELISA gave 100% positive results (23/23). The clinical signs ranged between splenomegaly & ascitis in 4.34% for each up to hepatomegaly in 73.91% & mild fever in 78.26% but 2 cases were asymptomatic. Mild eosinophilia and moderate anaemia were recorded with means of 11 +/- 5.8 and 10 +/- 1.3 respectively. Mean serum bilirubin was not elevated (0.91 +/- 0.51 gm/dl). Liver function tests (AST within normal range in all cases; <40 unit/ml but one patient had ALT above normal; >45 unit/ml). PMID- 18383808 TI - The effect of the host blood on the biology of the sand fly, Phlebotomus papatasi (Diptera: Psychodidae) under laboratory conditions. AB - The effect of the blood of human, Guinea pig and hamster on the different biological aspects of phlebotomid sand fly, Phlebotomus papatasi under laboratory conditions of 28 +/- 2 degrees C, 75 +/- 5% RH & a 14:10 hr. (L:D) photoperiod regime was evaluated. Man was the most preferable host followed by hamster, and then Guinea pig. Human blood gave the highest yield of eggs (mean: 65.15 eggs/female), highest hatchability (mean: 96.23%), shortest egg incubation period (mean: 7.37 days), shortest larval duration (mean: 22.59 days), shortest adult emergence period (mean: 39.73 days) and highest productivity of the adult progeny (mean; 81.75%). As human blood can not be currently used for laboratory rearing and colonization of sand flies, the hamster may be chosen as source of a substituted blood for the long term colonization of P. papatasi. PMID- 18383809 TI - Zoonotic fascioliasis in donkeys: ELISA (Fges) and postmortum examination in the Zoo, Giza, Egypt. AB - In Egypt, zoonotic fascioliasis is increasing. In this study, postmortum examination of 88 donkeys used as gargantuan meal in the Zoo at Giza revealed hepatic fascioliasis in 15 (17.05%). The serum examination for anti-Fasciola antibodies by ELISA showed positivity in 12/15 with crude worm antigen, and positivity in 14/15 with locally prepared Fasciola excretory-secretory (Fges) antigen. The zoonotic role of animal fascioliasis was discussed. PMID- 18383811 TI - Brain plasticity and tumors. AB - Brain plasticity is the potential of the nervous system to reshape itself during ontogeny, learning or following injuries. The first part of this article reviews the pathophysiological mechanisms underlying plasticity at different functional levels. Such plastic potential means that the anatomo-functional organization of the brain in humans, both physiological and pathological, has flexibility. Patterns of reorganization may differ according to the time-course of cerebral damage, with better functional compensation in more slowly growing lesions. The second part of this review analyzes the interactions between tumor growth and brain reshaping, using non-invasive (neuroimaging) and invasive (electrophysiological) methods of functional mapping. Finally, the therapeutic implications provided by a greater understanding of these mechanisms of cerebral redistribution are explored from a surgical point of view. Enhanced preoperative prediction of an individual's potential for reorganization might be integrated into surgical planning and preserving quality of life through tailored rehabilitation programmes to optimize functional recovery following resection of a brain tumor. PMID- 18383812 TI - Tumor-biology and current treatment of skull-base chordomas. AB - Chordomas are rare, slow growing tumors of the axial skeleton, which derive from the remnants of the fetal notochord. They can be encountered anywhere along the axial skeleton, most commonly in the sacral area, skull base and less commonly in the spine. Chordomas have a benign histopathology but exhibit malignant clinical behavior with invasive, destructive and metastatic potential. Genetic and molecular pathology studies on oncogenesis of chordomas are very limited and there is little known on mechanisms governing the disease. Chordomas most commonly present with headaches and diplopia and can be readily diagnosed by current neuroradiological methods. There are 3 pathological subtypes of chordomas: classic, chondroid and dedifferentiated chordomas. Differential diagnosis from chondrosarcomas by radiology or pathology may at times be difficult. Skull base chordomas are very challenging to treat. Clinically there are at least two subsets of chordoma patients with distinct behaviors: some with a benign course and another group with an aggressive and rapidly progressive disease. There is no standard treatment for chordomas. Surgical resection and high dose radiation treatment are the mainstays of current treatment. Nevertheless, a significant percentage of skull base chordomas recur despite treatment. The outcome is dictated primarily by the intrinsic biology of the tumor and treatment seems only to have a secondary impact. To date we only have a limited understanding this biology; however better understanding is likely to improve treatment outcome. Hereby we present a review of the current knowledge and experience on the tumor biology, diagnosis and treatment of chordomas. PMID- 18383813 TI - The influence of genetics on intracranial aneurysm formation and rupture: current knowledge and its possible impact on future treatment. AB - The etiology of intracranial aneurysm formation and rupture remains mostly unknown, but lately several studies have increasingly supported the role of genetic factors. In reports so far, genome-wide linkage studies suggest several susceptibility loci that may contain one or more predisposing genes. Depending on the examined ethnic population, several different non-matching chromosomal regions have been found. Studies of several candidate genes report association with intracranial aneurysms. To date, no single gene has been identified as responsible for intracranial aneurysm formation or rupture. In addition to the well-published environmental factors, such as alcohol intake, hypertension and smoking, only the recent progress in molecular genetics enables us to investigate the possible genetic determinants of this disease. Although a familial predisposition is the strongest risk factor for the development of intracranial aneurysms, the mode of Mendelian inheritance is uncertain in most families. Therefore, multiple genetic susceptibilities in conjunction with the environmental factors are considered to act together in the disease's etiology. Accordingly, researchers performed linkage studies and case-control association studies for the genetic analysis and have identified several genes to be susceptible to intracranial aneurysms. The identification of susceptible genes may lead to the understanding of the mechanism of formation and rupture and possibly lead to the development of a pharmacological therapy. Furthermore, should it be possible to identify a genetic marker associated with an increased risk of formation and rupture of an intracranial aneurysm, the necessity for screening and urgency of treatment could be determined more easily. In this review we summarize the current knowledge of intracranial aneurysm genetics and also discuss the method to detect the causalities. In view of the recent advances made in this field, we also give an outlook on possible future genetically engineered therapies, whose development are well underway. PMID- 18383814 TI - Extended endoscopic endonasal approach to the midline skull base: the evolving role of transsphenoidal surgery. AB - The evolution of the endoscopic endonasal transsphenoidal technique, which was initially reserved only for sellar lesions through the sphenoid sinus cavity, has lead in the last decades to a progressive possibility to access the skull base from the nose. This route allows midline access and visibility to the suprasellar, retrosellar and parasellar space while obviating brain retraction, and makes possible to treat transsphenoidally a variety of relatively small midline skull base and parasellar lesions traditionally approached transcranially. We report our current knowledge of the endoscopic anatomy of the midline skull base as seen from the endonasal perspective, in order to describe the surgical path and structures whose knowledge is useful during the operation. Besides, we describe the step-by-step surgical technique to access the different compartments, the "dangerous landmarks" to avoid in order to minimize the risks of complications and how to manage them, and our paradigm and techniques for dural and bony reconstruction. Furthermore, we report a brief description of the useful instruments and tools for the extended endoscopic approaches. Between January 2004 and April 2006 we performed 33 extended endonasal approaches for lesions arising from or involving the sellar region and the surrounding areas. The most representative pathologies of this series were the ten cranioparvngiomas, the six giant adenomas and the five meningiomas; we also used this procedure in three cases of chordomas, three of Rathke's cleft cysts and three of meningo-encephaloceles, one case of optic nerve glioma, one olfactory groove neuroendocrine tumor and one case of fibro-osseous dysplasia. Tumor removal, as assessed by post-operative MRI, revealed complete removal of the lesion in 2/6 pituitary adenomas, 7/10 craniopharyngiomas, 4/5 meningiomas, 3/3 Rathke's cleft cyst, 3/3 meningo-encephalocele. Surgical complications have been observed in 3 patients, two with a craniopharyngioma, one with a clival meningioma and one with a recurrent giant pituitary macroadenoma involving the entire left cavernous sinus, who developed a CSF leak and a second operation was necessary in order to review the cranial base reconstruction and seal the leak. One of them developed a bacterial meningitis, which resolved after a cycle of intravenous antibiotic therapy with no permanent neurological deficits. One patient with an intra-suprasellar non-functioning adenoma presented with a generalized epileptic seizure a few hours after the surgical procedure, due to the intraoperative massive CSF loss and consequent presence of intracranial air. We registered one surgical mortality. In three cases of craniopharyngioma and in one case of meningioma a new permanent diabetes insipidus was observed. One patient developed a sphenoid sinus mycosis, cured with antimycotic therapy. Epistaxis and airway difficulties were never observed. It is difficult todav to define the boundaries and the future limits of the extended approaches because the work is still in progress. Such extended endoscopic approaches, although at a first glance might be considered something that everyone can do, require an advanced and specialized training. PMID- 18383815 TI - Management of brachial plexus injuries. AB - Most brachial plexus lesions are traction injuries sustained during birth, but in adolescents and older people they are usually caused by traffic accidents or following a fall in the home. A minority are the result of penetrating injury after civilian assault or trauma encountered during wartime. Birth palsy cases (obstetric brachial plexus palsy) and the remaining cases (traumatic briachial plexus palsy) are viewed differently with regard to treatment and outcome and so these two groups are usually discussed in separate chapters. In this paper we treat both groups in parallel because as far as primary (= nerve) surgery is concerned, many treatment problems and solutions are present in both groups and are therefore comparable. PMID- 18383816 TI - Surgical anatomy of the jugular foramen. AB - The jugular foramen (JF) is a canal that makes communication between the posterior cranial fossa and the upper neck for one third of the cranial nerves and for the main venous channel of the brain. From a lateral view, the JF is protected by multiple layers of muscles and by the outer surface of the petrous bone. Surgical exposure of the JF is usually justified by the removal of benign tumors that grow in this region. In the first part of the present study we describe the surgical anatomy of the JF Then, we detail the relevant points of a stepwise surgical progression of three lateral skull base approaches with a gradual level of exposure and invasiveness. The infralabyrinthine transsigmoid transjugular-high cervical approach is a conservative procedure that associates a retrolabyrinthine approach to a lateral dissection of the upper neck, exposing the sinojugular axis without mobilization of the facial nerve. In the second step, the external auditory canal is transsected and the intrapetrous facial nerve is mobilized, giving more exposure of the carotid canal and middle ear cavity. In the third step, a total petrosectomy is achieved with sacrifice of the cochlea, giving access to the petrous apex and to the whole course of the intrapetrous carotid artery. Using the same dissection of the soft tissues from a lateral trajectory, these three approaches bring solutions to the radical removal of distinct tumor extensions. While the first step preserves the facial nerve and intrapetrous neurootologic structures, the third one offers a wide but more aggressive exposure of the JF and related structures. PMID- 18383817 TI - Inhibition of both mesothelioma cell growth and Cdk4 activity following treatment with a TATp16INK4a peptide. AB - Disruption of the 9p21 locus is common in mesothelioma and leads to loss of both the p16INK4a and the p14ARF gene products. This study tested the hypothesis that reexpression of p16INK4a carried out using the TAT delivery system that carries the protein transduction domain of the HIV TAT will result in mesothelioma cell death. MATERIALS AND METHODS: A synthetic TATp16INK4a peptide and a charge matched control were transduced into mesothelioma cells in vitro and in vivo. Cells were assayed for Cdk4 inhibition, cell cycle arrest, and cell death. RESULTS: Treatment of mesothelioma cells with TATp16INK4a for 48 hours resulted in cell death. Apoptosis and G1 cell cycle arrest was also observed. Following transduction of cells with TATp16INK4a there was complete but transient hypophosphorylation of pRb. Similar effects were observed in mesothelioma xenografts. CONCLUSION: Therapeutic strategies which introduce either TATp16INK4a peptide, or small molecule mimetic, could be an effective strategy for mesothelioma treatment. PMID- 18383818 TI - Cell cycle perturbation and acquired 5-fluorouracil chemoresistance. AB - Acquired chemoresistance is one of the obstacles for success of 5-fluorouracil (5 FU)-based cancer chemotherapy. Some molecular mechanisms of acquired 5-FU resistance are still unknown. We have recently demonstrated down-regulation of a group of cell cycle related genes in acquired 5-FU resistant human cancer cell lines. In this study, the bivariate distribution of propidium iodide versus BrdU in acquired 5-FU resistant colon (H630R10) and breast (T47DFU2.5) cancer cell lines was compared with their parental cell lines using flow cytometric analysis. The resistant cell lines showed significantly lower labelling index (T47DFU2.5) and cell cycle delay in G1 and G1/S boundary and prolonged DNA synthesis time (H630R10). Both resistant cell lines demonstrated significantly prolonged potential doubling time (Tpot). The protein expression levels of some G1 and S phase transition-related genes were also analysed by Western blot. CDK2 protein and Thr-160 phosphorylated CDK2 were remarkably reduced in the resistant cell lines. Cyclin D3 and cyclin A were also decreased in the resistant cells. Total pRB expression was unaltered but hypophosphorylation of pRB (Ser780, Ser795 and Ser807/811) was detected in the resistant cancer cells. Our data suggest that there may be a slow down in cell cycle traverse preventing incorporation of 5-FU metabolites into DNA and also providing cancer cells with sufficient time to correct the mis-incorporated nucleotides. The cell cycle perturbation may be involved in acquired 5-FU resistance. PMID- 18383819 TI - Proliferation and programmed cell death: role of p53 protein in high and low grade astrocytoma. AB - p53 is a cell cycle regulator that has been well-recognized as the key molecule that triggers the induction and the control of cell proliferation and apoptosis in a wide variety of tumours, including astrocytoma. Apoptosis and proliferation are two processes intimately coupled, that occur simultaneously in tumour tissue. Previous studies of the correlations between proliferation and apoptotic index with p53 expression in astrocytic tumours have remained inconclusive. The aim of this study was to investigate the correlation of p53 expression with the apoptotic index (AI) and the cell proliferation index (PI) in pilocytic astrocytoma (PA) and glioblastoma multiforme (GBM). A correlation of p53 expression with AI and PI was found in pilocytic astrocytoma but not in glioblastoma, probably because of the mutated p53 phenotype in the latter. PMID- 18383820 TI - Chemotherapy-induced bystander effect in response to several chloroethylnitrosoureas: an origin independent of DNA damage? AB - Chloroethylnitrosourea (CENU) chemotherapy is used for the treatment of melanoma tumors. The main mechanism of action of this anticancer agent is via DNA damage. We recently showed in murine experiments using a parental double B16 melanoma tumor model that, after treatment of primary tumors with cystemustine (CENU agent), untreated secondary tumors exhibited growth inhibition and metabolism disorders. The response of secondary untreated tumor was called the chemotherapy induced bystander effect. To see whether chemotherapy-induced bystander effects were induced with other members of the CENU family, we compared three CENU(s) used in melanoma treatment: cystemustine, carmustine and fotemustine. Our results demonstrate that fotemustine, like cystemustine, but not carmustine induced a protective effect against secondary untreated tumors including alterations in phospholipid derivative and glutathione which are the metabolic signature of the bystander effect. From these data we may conclude that DNA damage to the primary tumor is not sufficient to explain chemotherapy-induced bystander effects. PMID- 18383821 TI - Identification of glutathione sulfotransferase-pi (GSTP1) as a new resveratrol targeting protein (RTP) and studies of resveratrol-responsive protein changes by resveratrol affinity chromatography. AB - BACKGROUND: Resveratrol shows chemopreventive and other biological affects in in vitro and some animal studies. The bioactivities of resveratrol may be attributed to qualitative and quantitative differences in its cell-type-specific interaction and binding with its cellular targets, denoted as resveratrol targeting proteins (RTPs). MATERIALS AND METHODS: To isolate RTPs, resveratrol was linked to epoxy activated agarose generating an affinity platform to allow the isolation, purification, and characterization of distinct RTPs from cultured prostate cancer cell extracts. RESULTS: Glutathione sulfotransferase-pi (GSTP1) and estrogen receptor-beta (ER-beta) were found to be new RTPs. Resveratrol affinity chromatography was shown to be an easy method for analyzing resveratrol responsive protein changes in the androgen-dependent LNCaP cells. CONCLUSION: Resveratrol affects cellular functions at multiple levels, ranging from interaction with detoxification enzymes, such as GSTP1 and transcription by targeting factors such as ER-beta. PMID- 18383822 TI - Induction of necrosis in human myeloma cells by kigamicin. AB - BACKGROUND: Kigamicin (KGM) is a novel compound derived from Actinomycetes that was originally reported to induce necrosis in pancreatic cancer cells only under nutrient-starved conditions via inhibition of PI3-kinase. The effects of KGM on myeloma cells were investigated. MATERIALS AND METHODS: Cytotoxic activity was quantified using WST8 assay. Necrosis was determined by Annexin V/PI staining. Regulatory protein levels were assessed by Western blot. LY294002 was utilized as a PI3-kinase inhibitor. RESULTS: KGM induced necrosis in myeloma cells in nutrient rich conditions with a CC50 of approximately 100 nM. KGM did not induce necrosis in normal lymphocytes. Cyclin D1, p21, p-AKT and p-ERK were inhibited by KGM while LY294002 did not inhibit cell death by KGM. A melphalan-resistant myeloma cell line was more susceptible to KGM than the melphalan-sensitive parental cell line. CONCLUSION: KGM-induced necrosis in myeloma cells even at very low concentration. The present data warrant further investigation into the use of KGM as a potential therapeutic agent for multiple myeloma. PMID- 18383823 TI - The utility of vitamin K3 (menadione) against pancreatic cancer. AB - BACKGROUND: To evaluate the efficacy of vitamin K3 (VK3) against pancreatic cancer, the molecular mechanism of VK3 or gemcitabine (GEM)-induced inhibition of proliferation was characterized. MATERIALS AND METHODS: The cell viability was determined using the 3-[4,5-dimethylthiazol]-2,5-diphenyl tetrazolium bromide (MTT) test method. The expressions of cellular proteins were evaluated by Western blot analysis. For morphological studies of the in vivo transplanted cancer cells, the tissues were stained with hematoxylin and eosin. RESULTS: The IC50 of VK3 for pancreatic cancer cells was calculated for 42.1 +/- 3.5 microM. Western blot analysis showed that VK3 induced rapid phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) 30 minutes after application. ERK but not JNK phosphorylation was maintained for at least 12 hours. Activation of apoptosis by VK3, as shown by molecular weight shifts of the pro-activated 32-kDa form of caspase-3 and poly(ADP-ribose)polymerase (PARP) cleavage of the 112-kDa form, was found. Treatment with the thiol antioxidant, L cysteine (>0.2 mM), completely abrogated the VK3-induced phosphorylation of ERK, but not the JNK, and inhibition of proliferation. A caspase-3 inhibitor antagonized caspase-3 activation, but had no inhibitory effect on the proliferative activity of VK3. GEM at concentrations >0.1 microg/ml was found to inhibit cell proliferation after 24 hours. GEM also induced phosphorylation of JNK, activation of caspase-3 and accumulation of cyclin B1. Local application of VK3 was found to induce extensive tumor tissue necrosis, but slight hematemesis without necrosis was observed 48 hours after GEM injection. In Western blot, ERK but not JNK phosphorylation, was clearly detected in response to VK3 injection into the tumor tissue. CONCLUSION: The action of VK3 may lead to a favorable outcome against pancreatic cancer, and the detection of ERK phosphorylation in the tissue is important for predicting this effect. PMID- 18383824 TI - Adenovirus vector-mediated gene therapy using iodized oil esters for hepatocellular carcinoma in rats. AB - BACKGROUND: When gene therapy is performed for malignant tumors, gene transfer efficiency and selectivity are extremely important. The usefulness of gene therapy by intraarterial injection of an adenovirus vector with iodized oil esters (IOEs) for hepatocellular carcinoma (HCC) was studied. MATERIALS AND METHODS: HCC was induced in rats with diethyl nitrosamine and phenobarbital, after which either adenovirus vector expressing the herpes simplex virus thymidine kinase (AxCAHSVtk) and IOEs or AxCAHSVtk alone was injected through the hepatic artery. On postoperative days 2, 4 and 6, gancyclovir was injected into the peritoneum; blood sampling was performed on day 7. RESULTS: Aspartate aminotransferase and alanine aminotransferase levels in the AxCAHSVtk with IOEs group were lower than in the AxCAHSVtk alone group (p = 0.0274, p = 0.0323). However, the survival rate was not significantly different between groups (p = 0.7122). CONCLUSION: Intra-arterial injection of an adenovirus vector with IOEs can result in cancer-selective but not effective gene therapy for HCC. PMID- 18383825 TI - Effect of pO2 on antitumor drug cytotoxicity on MDR and non-MDR variants selected from the LoVo metastatic colon carcinoma cell line. AB - Two chemosensitive cell lines, LoVo-fusoid (LoVo-f) and LoVo-small cells (LoVo sc) were derived from the original LoVo cell line. These two variants and the multidrug-resistant (MDR) cell line LoVo-Dox were screened for various properties. In non-permeabilized cells, only LoVo-sc showed mucin-2 staining whereas labelling was positive in all permeabilized cell lines. As shown by electron microscopy screening and by relative resistance to trypsin detachment, only LoVo-sc cells showed strong mucus secretion. All three cell lines displayed strong staining for P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP) and lung-resistance-related protein (LRP) in different locations according to the drug resistance state. The three cell lines showed intracellular labelling of LRP and MRP. The sensitive cells showed P-gp in a large perinuclear ring and in the cytoplasm, but little (LoVo-sc cells) or no staining (LoVo-f cells) was shown at the plasma membrane level. For the Lovo-Dox cells, P-gp was located in the plasma membrane, in cellular anchorages and in the cytoplasm as well. Cell resistance against antineoplastic agents often results from mobilization of various factors, the modulation of which is linked to the culture conditions. As most of the protocols utilize cells growing in (air + 5-10% CO2) atmosphere e.g. 20% O2, balance of the respective participants in the MDR multi modal mechanism may not be representative of the in vivo situation and may lead to erratic pharmacological response. Indeed, cells within solid tumours were exposed to low pO2, most of them being under hypoxic condition (0.1-5% O2). In the absence of anticancer drugs, all LoVo cell lines grew notably faster at 20% O2 than at 5% O2. Moreover, respective sensitivities of both non-MDR variants to doxorubicin were altered according the pO2. Whatever the pO2 was, virtually none of the antioxidants tested affected the cytotoxic activity of doxorubicin for the three cell lines. By contrast, trolox showed a strong inhibitory effect on doxorubicin activity. These results underline the importance of evaluating the role of hypoxia on the cytotoxic effect of chemotherapeutic agents used either as single drugs or in combination therapy. PMID- 18383826 TI - Feasibility of induction heating using a sintered MgFe2O4 needle for minimally invasive breast cancer therapy. AB - BACKGROUND: This study investigated a novel approach for tumor ablation therapy using an alternating magnetic field combined with a sintered MgFe2O4 needle. This method differs from radiofrequency ablation (RFA) by dielectric heating with regard to the heating mechanism and improves some weak points of these conventional thermotherapies. MATERIALS AND METHODS: Nude mice mimicking human breast cancer BT474 were treated using this method. The extent of tumor death was assessed after ablation. RESULTS: Staining with hematoxylin and eosin showed gradual expansion of the pyknotic area until 48 h after ablation. Nicotinamide adenine dinucleotide diaphorase staining also showed complete tumor death by 48 h after treatment. The ablation area was well controlled and reablation was not necessary. The tumor could be completely controlled using this method without any risk of skin burn. CONCLUSION: This novel ablation therapy appeared to be more effective and less invasive for treatment of breast cancer treatment than RFA. PMID- 18383827 TI - Volume-sensitive Cl(-) channel as a regulator of acquired cisplatin resistance. AB - The platinum-based drug cisplatin (cis-diamminedichloroplatinum (II)) is widely used in cancer therapy. However, cancer cells can develop resistance after exposure to cisplatin. Recently, many studies have pointed to the involvement of plasma membrane ion channels in a cell's response to cisplatin. Our group has found that pretreatment with cisplatin enhanced the activity of volume-sensitive C-channels in human epidermoid cancer KB cells; cisplatin-resistant KCP-4 cells derived from KB cells, on the other hand, lacked functional expression of these channels. This suggests that the activity of volume-sensitive Cl(-) channels is an important factor in determining the sensitivity of cancer cells to cisplatin. Furthermore, when volume-sensitive Cl(-) channel function was partially restored in cisplatin-resistant KCP-4 cells treated with a histone deacetylase inhibitor, KCP-4 cells exhibited a restoration of sensitivity to cisplatin; this increased sensitivity was inhibited by a volume-sensitive Cl(-) channel blocker. We therefore propose that impaired activity of the volume-sensitive Cl(-) channel is involved in the acquired cisplatin resistance of these cancer cells. In this review, we will outline the relationship between volume-sensitive Cl(-) channels, cisplatin-induced apoptosis, and cisplatin resistance. Activating the volume sensitive outwardly-rectifying Cl(-) channel may be a new strategy in treating clinical cisplatin resistance. PMID- 18383828 TI - Delta(9)-tetrahydrocannabinol inhibits 17beta-estradiol-induced proliferation and fails to activate androgen and estrogen receptors in MCF7 human breast cancer cells. AB - BACKGROUND: Delta(9)-tetrahydrocannabinol (THC) exerts palliative effects in cancer patients, but produces adverse effects on the endocrine and reproductive systems. Experimental evidence concerning such effects is controversial. Whether THC exhibits estrogenic or androgenic activity in vitro was investigated. MATERIALS AND METHODS: Estrogenic effects of THC were analyzed in vitro by measuring the proliferation of estrogen-sensitive MCF7 cells. Androgenic activity was investigated by the A-Screen assay that measures androgen-dependent inhibition of proliferation of the androgen receptor (AR)-positive human mammary carcinoma cell line, MCF7-AR1. RESULTS: In contrast to 17beta-estradiol, included as positive control with an EC50 value (concentration required for 50% of maximal 17beta-estradiol-induced proliferation) of 1.00 x 10(-12) M, THC failed to induce cell proliferation in the MCF7 cell line at concentrations between 10(-13) and 10(-4) M. THC inhibited 17beta-estradiol-induced proliferation in wild-type MCF7 and MCF7-AR1 cells, with an IC50 value of 2.6 x 10(-5) M and 9 x 10(-6) M, respectively. CONCLUSION: THC failed to act as an estrogen, but antagonized 17beta-estradiol-induced proliferation. This effect was independent of the AR expression level. PMID- 18383829 TI - Parasporin-1Ab, a novel Bacillus thuringiensis cytotoxin preferentially active on human cancer cells in vitro. AB - The non-insecticidal Bacillus thuringiensis strain B0195 produces parasporin (PS) whose cytocidal activity is preferential for human cancer cells. This study identified two ps genes, ps1Aa3 and ps1Ab1, from the strain B0195. The former gene was 2,169-bp long, encoding an 81 kDa protein (PSLAa3) whose aminoacid sequence was 100% identical to that of the reference protein PS1Aa1. The latter gene was 2,178-bp long, encoding a novel protein of 82 kDa, PS1Ab1, whose sequence was 86.4% identical to that of PS1Aa1. The recombinant protein of PS1Ab1, synthesized in transformed B. thuringiensis cells, induced marked cytopathy in HeLa cells (human uterus cervix cancer cells) upon proteolytic activation. The cytopathy was characterized by cell-ballooning, followed by gradual cell-shrinking. Unlike HeLa cells, non-cancer UtSMC cells (human uterine smooth muscle-cells) were not susceptible to PS1Ab1. PMID- 18383830 TI - Polymorphisms of death pathway genes FAS and FASL and risk of premalignant gastric lesions. AB - BACKGROUND: Tumorigenesis is a multistep process that begins with the abrogation of normal controls of apoptosis and cell proliferation, and the FAS receptor ligand system is a key regulator of apoptosis. The aim of this study was to investigate whether functional polymorphisms of death pathway genes (FAS and FASL) are associated with the development of gastric atrophy and intestinal metaplasia. PATIENTS AND METHODS: Genotypes in the promoter regions of the FAS ( 1377G/A and -670A/G) and FASL (-844T/C) genes of 101 healthy individuals and 86 gastric cancer patients were determined by PCR-RFLP. Additionally, gastric histological changes were examined according to the updated Sydney System. RESULTS: The carriage of FASL -844C allele significantly increased the risk of atrophy in the gastric corpus, with an adjusted odds ratio (OR) of 5.0 [95% confidence interval (CI), 1.5-6.8]. There were no gene-gene interactions among FASL -844T/C, FAS -1377G/A and FAS -670A/G polymorphisms in developing premalignant gastric lesions. In the 109 individuals with Helicobacterpylori infection, carrying the FAS -1377A allele was a protective factor for developing intestinal metaplasia in the antrum (OR, 0.3; 95% CI, 0.1-0.9), while carrying the FASL -844C allele was a risk factor for developing gastric atrophy in the corpus (OR, 9.4; 95% CI, 1.7-53.4). CONCLUSION: FAS and FASL genotypes of the hosts are important determinants in the pathogenesis of gastric atrophy and intestinal metaplasia in H. pylori-infected individuals. PMID- 18383831 TI - A new peritoneal carcinomatosis model in cyclosporine immunosuppressed rats. AB - Colorectal cancer (CRC) cells (HT29) were injected into the intraperitoneal (i.p.) cavity of immunosuppressed male Sprague Dawley rats. Immunosuppression was achieved via a daily i.p. injection of cyclosporine 35 mg/kg/day. Three groups of rats (n = 6 per group) were injected with 20, 50 or 100 million HT29 cells. All animals developed tumours indicating that this animal model may prove useful for future research in human colorectal carcinomatosis. PMID- 18383832 TI - Survivin expression in glioblastomas correlates with proliferation, but not with apoptosis. AB - BACKGROUND: Survivin is expressed in proliferating tissues and in tumors. It is a member of the inhibitory apoptosis protein (IAP) family known to regulate mitosis and to inhibit apoptosis. It has therefore been regarded as a target for therapies. In malignant gliomas it increases with malignancy, even though in glioblastomas it does not seem to correlate with outcome. MATERIALS AND METHODS: Survivin was immunohistochemically studied in 39 selected viable glioblastoma areas belonging to 20 cases which were assayed for apoptosis, using a TUNEL assay, caspase-3, poly(ADP-ribose)polymerase 1 (PARP-1), Bid (BH3-interacting domain death agonist) and with the proliferation index Ki-67/MIB-1 and mitotic index (MI). RESULTS: A positive linear correlation was found between the survivin labelling index (LI) and the Ki-67/MIB-1 LI and MI. No inverse correlation was found with apoptosis. CONCLUSION: This double behavior can be attributed to mechanisms mediating survivin activity, either as a mitosis regulator and apoptosis inhibitor, and should be taken into account in therapeutic strategies using survivin. PMID- 18383833 TI - Unlocking pathology archives for microRNA-profiling. AB - BACKGROUND: MicroRNAs (miRNAs) are approximately 22 nucleotide long, non-coding RNAs that regulate gene expression by binding to the 3'-untranslated region of target mRNAs and also a variety of cellular processes. It has recently been established that dysregulation of miRNA expression can be detected in the majority of human cancers. A variety of high-throughput screening methods has been developed to identify dysregulated miRNA species in tumours. For retrospective clinical studies formalin-fixed, paraffin-embedded (FFPE) tissue is the most widely used material. MATERIALS AND METHODS: The miRNA expression profiles of freshly frozen (CRYO) and FFPE tissues of seven tonsil and four liver samples were compared, using a qPCR-based assay, profiling 157 miRNA species. RESULTS: The significance of miRNA-profiles was barely influenced by FFPE treatment in both tissues and the variance induced by FFPE treatment was much smaller than the variance caused by biologically based differential expression. CONCLUSION: FFPE material is well suited for miRNA profiling. PMID- 18383834 TI - EpCAM overexpression is associated with high-grade urothelial carcinoma in the renal pelvis. AB - BACKGROUND: EpCAM serves as an attractive target for immunotherapy due to its expression on the surface of most epithelial cancer cells. Urothelial carcinoma of the renal pelvis (RP-UC) comprises 2.4-4.6% of tumors of the lower urinary tract. To assess the expression of EpCAM in RP-UC a retrospective study was performed. PATIENTS AND METHODS: Tumor tissue from 42 patients with RP-UC was selected from the archives of the Institute of Pathology, Medical University of Innsbruck, Austria. EpCAM expression was demonstrated by immunohistochemistry using the mouse monoclonal antibody ESA. RESULTS: EpCAM overexpression was significantly associated with high grade and invasive behaviour (p = 0.014 and p = 0.029) and the presence of lymph node metastases (p = 0.031), but not with the extent of nodal involvement (p = 0.12). CONCLUSION: In RP-UC, EpCAM overexpression is associated with an aggressive tumor phenotype. The association of EpCAM overexpression with the presence of lymph node metastasis may be of prognostic and therapeutic relevance. PMID- 18383835 TI - Effects of alpha-santalol on proapoptotic caspases and p53 expression in UVB irradiated mouse skin. AB - BACKGROUND: Cancer chemoprevention by naturally occurring agents, especially phytochemicals, minerals and vitamins has shown promising results against various malignancies in a number of studies both under in vitro and in vivo conditions. One such phytochemical, alpha-santalol, a major component of sandalwood oil, is effective in preventing skin cancer in both chemically and UVB-induced skin cancer development in CD-1, SENCAR and SKH-1 mice; however, the mechanism of its efficacy is not fully understood. The objective of the present investigation was to study the effects of alpha-santalol on apoptosis proteins and p53 in UVB induced skin tumor development in SKH-1 mice to elucidate the mechanism of action. MATERIALS AND METHODS: Female SKH-1 mice were divided into two groups: Group 1, which served as control received topical application of acetone (0.1 ml) one hour before UVB treatment; Group 2 received alpha-santalol (0.1 ml, 5% w/v in acetone, topical) one hour prior to UVB treatment. UVB-induced promotion was continued for 30 weeks. RESULTS: Pre-treatment with alpha-santalol one hour prior to UVB exposure significantly (p < 0.05) reduced tumor incidence and multiplicity, and resulted in a significant (p < 0.05) increase in apoptosis proteins, caspase-3 and -8 levels and tumor suppressor protein, p53. CONCLUSION: These results suggest that alpha-santalol prevents skin cancer development by inducing proapoptotic proteins via an extrinsic pathway and increasing p53. PMID- 18383836 TI - Wilms' tumor 1 protein and focal adhesion kinase mediate keratinocyte growth factor signaling in breast cancer cells. AB - BACKGROUND: Keratinocyte growth factor (KGF) has been shown to induce breast cancer metastasis in animal models. cDNA microarrays have revealed that KGF increased Wilms tumor 1 (WT1) and focal adhesion kinase (FAK) expression in breast cancer cells. The role of WT1 and FAK in KGF signaling was investigated. MATERIALS AND METHODS: A cell culture wounding model was used to study the effects of WT1 and FAK down-regulation on KGF-induced proliferation and motility in breast cancer cells. RESULTS: WT1 down-regulation inhibited KGF-mediated proliferation and motility of breast cancer cells, while FAK down-regulation inhibited proliferation, but had no significant effect on cell motility. WT1 down regulation, but not FAK down-regulation, led to Erk1,2 inactivation. CONCLUSION: KGF-mediated signaling employs WT1 and FAK to regulate breast cancer cell proliferation and motility and may represent therapeutic targets for the prevention of breast cancer progression. PMID- 18383837 TI - Overexpression of the mitotic spindle assembly checkpoint genes hBUB1, hBUBR1 and hMAD2 in thyroid carcinomas with aggressive nature. AB - BACKGROUND: The mitotic spindle assembly checkpoint (MSAC) genes are responsible for preventing chromosome missegregation. MSAC gene expressions have been reported to be associated with tumor cell proliferation or unfavorable cancer behavior. The present study was conducted to preliminary investigate the MSAC gene expressions in thyroid neoplasms. MATERIALS AND METHODS: Expression levels of MSAC genes (hBUB1, hBUBR1, hBUB3 and hMAD2) were evaluated in 9 follicular thyroid adenomas (FAs), 9 follicular thyroid carcinomas (FTCs), 21 papillary thyroid carcinomas (PTCs), 5 anaplastic (undifferentiated) thyroid carcinomas (ATCs) and 3 adjacent normal thyroid tissues (NTs) by real-time quantitative RT PCR. These gene expressions were compared between undifferentiated thyroid carcinomas (ATCs) and differentiated thyroid carcinomas (DTCs) and between advanced DTCs and non-advanced DTCs. DTCs included PTCs and FTCs. Advanced DTCs were defined as carcinoma with aggressive nature such as extrathyroid extension, distant metastasis, recurrence or death from the disease. RESULTS: MSAC gene expressions varied in different thyroid tumors and fell in the order of ATC, DTC (PTC and FTC), FA and NT Carcinomas had higher expression compared to adenoma or normal tissue. hBUB1, hBUBR1 and hMAD2 expressions in ATCs were significantly higher than those in DTCs (p<0.005). hBUBR1 and hMAD2 expressions in advanced DTCs were significantly higher than those in non-advanced DTCs (p<0.05). CONCLUSION: The MSAC genes were overexpressed in thyroid carcinomas with aggressive nature. Further studies are required to clarify the relationship between the MSAC gene expressions and thyroid cancer behavior. PMID- 18383838 TI - New method for colorectal cancer diagnosis based on SSCP analysis of DNA from exfoliated colonocytes in naturally evacuated feces. AB - BACKGROUND: Establishment of a sensitive, reliable and non-invasive method for the diagnosis of early-stage colorectal cancer is necessary, because colorectal cancer can be cured surgically if diagnosed early. The aim of the present study was to evaluate the feasibility of using PCR-SSCP (polymerase chain reaction single strand conformation polymorphism) analysis of the DNA extracted from the colonocytes isolated from naturally evacuated feces, in order to detect colorectal cancer. MATERIALS AND METHODS: Colonocytes exfoliated into feces, retrieved from 33 patients with colorectal cancer and 63 healthy volunteers, were analyzed for the presence of mutations in their DNA. The DNA extracted from the colonocytes was examined for mutations of the APC, K-ras and p53 genes using direct sequence analysis, and also subjected to PCR-SSCP analysis. RESULTS: Genetic alterations were detected in the colonocytes isolated from the feces of 12 out of 33 patients with colorectal cancer (36.4%) by direct sequence analysis. PCR-SSCP analysis using the same DNA samples revealed abnormal signals in 18 of the 33 patients (54.5%). However, 3 and 7 among the 63 healthy volunteers were also found to have abnormal genetic alterations by direct sequence and PCR-SSCP analysis, respectively. CONCLUSION: The present study indicated the feasibility of using PCR-SSCP analysis for the detection of mutations in the DNA extracted from colonocytes isolated from naturally evacuated fecal samples. PMID- 18383839 TI - Tumor-specificity and type of cell death induced by vitamin K2 derivatives and prenylalcohols. AB - Fourteen vitamin K2 (menaquinone (MK)-n, n = 1-14) and ten prenylalcohol derivatives (n = 1-10) with different numbers (n) of isoprenyl groups in the side chains were investigated for their cytotoxicity against nine human tumor cell lines and three human normal oral cells. Among the vitamin K2 derivatives, MK-2 (n = 2) showed the greatest cytotoxicity, followed by MK-1 (n = 1) and MK-3 (n = 3). MK-1, MK-2 and MK-3 showed the highest tumor-specific index (TS= > 2.0, 2.0 and > 1.7, respectively). Among the prenylalcohols, geranylgeraniol (GG) (n = 4) showed the highest cytotoxicity, followed by farnesol (n = 3) and geranylfarnesol (GF) (n = 3). GG showed the highest tumor-specificity (TS = 1.8), followed by farnesol (TS = > 1.4), GF (TS= > < 1.3). However, the tumor-specificity of MK-2 and GG was much lower than that of conventional chemotherapeutic agents. The human leukemic cell lines were the most sensitive, whereas the human glioblastoma cell lines were the most resistant to MK-2 and GG. MK-2 did not induce internucleosomal DNA fragmentation in either the human promyelocytic leukemia HL 60 or the human squamous cell carcinoma HSC-4 cell lines. GG induced marginal internucleosomal DNA fragmentation in the HL-60 cells, but not in the HSC-4 cells. Both MK-2 and GG did not induce the formation of autophagosomes, nor did they clearly change the intracellular concentration of three polyamines. Electron spin resonance (ESR) spectroscopy showed that only MK-1 (n = 1), as well as GGF (n = 7) and GFF (n = 8) which had lower cytotoxicity, produced radicals, suggesting the lack of connection between cytotoxicity and radical production. The present study demonstrates that the presence of 1,4-naphtoquinone structure (including alpha,beta-unsaturated ketones) in vitamin K2 derivatives confers on them the ability to induce non-apoptotic cell death. PMID- 18383840 TI - Expression of the serine protease hepsin and clinical outcome of human endometrial cancer. AB - BACKGROUND: Hepsin is a type II transmembrane serine protease originally identified in the human liver as a cDNA clone. Hepsin was found to be significantly overexpressed in cancer samples compared to matched various tissues (e.g. prostate, renal, ovarian carcinoma). The purpose of the present study was to examine hepsin expression and to evaluate its clinicopathological significance in endometrial cancer. PATIENTS AND METHODS: Hepsin expression was examined by immunohistochemisty in 34 cases with normal endometrium as a control, 11 cases with endometrial hyperplasia, and 128 cases with endometrioid adenocarcinoma. RESULTS: Hepsin expression was found to be significantly higher in endometrial cancer compared to normal endometrium and endometrial hyperplasia. High levels of hepsin expression were associated with advanced stage (p<0.001), high grade (p=0.002), depth of myometrial invasion (p<0.001), cervical involvement (p=0.007), lymph node metastasis (p=0.001), lymph vascular space (LVS) involvement (p=0.006), ovarian metastasis (p=0.002), and peritoneal cytology (p=0.03) of endometrial cancer. CONCLUSION: These findings indicate that hepsin protein expression could be an important indication for high risk of endometrial cancer. PMID- 18383841 TI - Preoperative serum vascular endothelial growth factor-C (VEGF-C) levels predict recurrence in patients with esophageal cancer. AB - BACKGROUND: Circulating vascular endothelial growth factor-C (VEGF-C) levels were measured in patients with esophageal cancer to assess the value of VEGF-C as a biomarker for predicting tumor recurrence. PATIENTS AND METHODS: Preoperative serum samples were acquired from 80 patients and healthy volunteers who served as normal controls. VEGF-C levels were assessed using enzyme-linked immunosorbent assay (ELISA). RESULTS: The preoperative serum VEGF-C level in patients with esophageal cancer was significantly higher than in healthy volunteers. Furthermore, patients with recurrence had significantly higher preoperative serum VEGF-C levels than patients without recurrence, and a high preoperative serum VEGF-C level was found to be an independent risk factor for recurrence, in addition to lymph node metastasis. CONCLUSION: Preoperative VEGF-C levels may reflect malignancy, such as lymph node metastasis, and predict recurrence in patients with esophageal cancer. Therefore, the preoperative VEGF-C level may be a useful biomarker for choice of multimodality therapy. PMID- 18383842 TI - Inhibition of NO production in LPS-stimulated mouse macrophage-like cells by trihaloacetylazulene derivatives. AB - The effect of 20 trihaloacetylazulene derivatives with one halogen atom, on nitric oxide (NO) production by mouse macrophage-like cells Raw 264.7 was investigated. 2-Methoxyazulenes and 2-ethoxyazulenes exhibited comparable cytotoxicity. Trichloroacetylazulenes generally exhibited higher cytotoxicity, as compared with the corresponding trifluoroacetylazulenes. Substitution of chloride, bromide or iodine at the C-3 position further enhanced their cytotoxicity. All of these compounds failed to stimulate the Raw 264.7 cells to produce detectable amounts of NO, but did inhibit NO production by LPS-activated Raw 264.7 cells to different extents. 1-Trichloroacetyl-2-methoxyazulene and 1 trichloroacetyl-2-ethoxyazulene, with less cytotoxic activity, inhibited NO production to the greatest extent, producing the highest selectivity index (SI) of >24.7 and >28.7, respectively. This was accompanied by the efficient inhibition of inducible NO synthase (iNOS) mRNA expression, but not by iNOS protein abundance. Electron spin resonance (ESR) spectroscopy showed that neither of these compounds produced radicals, nor scavenged NO, superoxide anion or diphenyl-2-picrylhydrazyl radicals. The present study suggests that the inhibitory effects of trifluoroacetylazulenes and trichloroacetylazulenes on NO production by activated macrophages might be derived from the perturbation of NO anabolism (inhibition of iNOS mRNA expression and possibly the inactivation of iNOS protein) rather than NO catabolism (NO scavenging). PMID- 18383843 TI - Pretreatment with S-1, an oral derivative of 5-fluorouracil, enhances gemcitabine effects in pancreatic cancer xenografts. AB - BACKGROUND: The systemic administration of gemcitabine (GEM) has been accepted as a standard treatment for patients with advanced pancreatic cancer. The major mediator of cellular uptake of GEM is the human equilibrative nucleoside transporter 1 (hENT1) whose expression is up-regulated by thymidylate synthase inhibitors, such as 5-fluorouracil (5-FU). S-1 is a novel oral derivative of the 5-FU prodrug tegafur combined with two modulators. Recent clinical trials have reported the promising effect of S-1 in pancreatic cancer. The purpose of this study was to evaluate the relationship between different schedules and the effects of GEM/S-1 combination therapy on pancreatic cancer xenograft models. MATERIALS AND METHODS: Human pancreatic tumor xenografts were prepared by subcutaneous implantation of MiaPaCa-2 into nude mice. Expression of hENT1 was determined by quantitative RT-PCR. GEM cellular uptake was determined using [3H] GEM. RESULTS: Significant increases in hENT1 expression and GEM cellular uptake were observed after S-1 treatment. Six different treatment schedules (no treatment, single agent of GEM or S-1, combination treatment with GEM either before, simultaneously or following administration of S-1) were compared. Significant tumor growth inhibition was observed in the mice treated with S-1 followed by GEM compared to either untreated mice or the mice treated with the other schedules. CONCLUSION: Based on the effects of S-1 on the uptake of GEM, S 1 should be used before GEM treatment. The GEM/S-1 combination therapy in patients with pancreatic cancer may be promising and should be tested in clinical trials. PMID- 18383844 TI - Human monocytes both enhance and inhibit the growth of human pancreatic cancer in SCID mice. AB - BACKGROUND: Monocytes/macrophages exhibit antitumour potential, but clinicopathological evidence suggests that they may both inhibit and enhance tumour growth. The purpose of this study was to determine the effect of monocytes on the growth of human pancreatic cancer (HPC-4) in severe combined immunodeficient (SCID) mice. MATERIALS AND METHODS: Freshly isolated human monocytes or CD14+ cells from cocultures with tumour cells were coengrafted, at various ratios, with HPC-4 cells into SCID mice. The tumour size and angiogenesis were determined. RESULTS: At a high ratio of monocytes to cancer cells the enhancement and, at a low ratio, the inhibition of tumour growth was observed. Multiple intratumoral applications of monocytes in large numbers also enhanced tumour growth. Deactivation of monocytes by a short pre-exposure to tumour cells in vitro before engraftment led to increased tumour growth. Monocytes used in large numbers and deactivated monocytes in low doses enhanced tumour-induced angiogenesis. CONCLUSION: Monocytes may both facilitate and suppress the growth of human tumours in SCID mice and both the number of monocytes, as well as the state of monocyte deactivation are critical for the final outcome of monocyte tumour interactions. PMID- 18383845 TI - Morphological events found at the invading edge of colorectal carcinomas in baboons. AB - BACKGROUND: Earlier studies at the growing edge of colorectal cancer (CRC) in humans and rats have shown dilated neoplastic glands, some with a thin layer of flattened tumor cells (FTCs), some lacking one or more groups of consecutive lining tumor cells (called glandular pores, GPs). MATERIALS AND METHODS: The characteristics of the neoplastic glands at the invading edge of CRCs were investigated in 39 baboons. A total of 190 neoplastic glands were studied in the 38 cases of glandular-forming adenocarcinomas. RESULTS: In the studied neoplastic glands FTCs or GPs were recorded in 44.7% (85 glands). FTCs were found in 9.5% (18 glands) and GPs in 35.3% (67 glands). Only 7.9% or 3 out of the 38 animals showed neoplastic glands with GPs in the bulk of the tumor. CONCLUSION: In similarity to colorectal adenocarcinomas in humans, flattened tumor cells and glandular pores were found at the invading tumor edge of colorectal adenocarcinomas in baboons. A possible mechanism of host invasion is proposed. PMID- 18383846 TI - Establishment of three novel human malignant pleural mesothelioma cell lines: morphological and cytogenetical studies and EGFR mutation status. AB - The incidence of mesothelioma is estimated to rise sharply worldwide, including Japan, in the next two decades. Molecular and proteomic studies are urgently required to elucidate the pathobiology of malignant mesothelioma. This paper describes the characterization of novel human malignant pleural mesothelioma cell lines representing the sarcomatoid, epithelioid and biphasic subtypes. MATERIALS AND METHODS: Established pleural effusion fluid cell lines were observed using phase-contrast microscopy and transmission electron microscopy. The immunoreactivity of the cells was evaluated using immunohistochemistiy, FACS analysis and Western blotting. The expression of SV40 large cell antigen and the EGFR mutation status were also analyzed. RESULTS: The cell lines had different morphological and immunophenotypic characteristics. All cell lines showed immunophenotypic marker expression of vimentin, mesothelin and N-cadherin, but no expression of CEA or E-cadherin. At the electron microscopic level, a cell surface rich in microvilli confirmed mesothelial origin of the cell lines. Karyotype analyses showed complex abnormalities in all cell lines. Neither EGFR mutations relevant to tyrosine kinase inhibitor responsiveness nor the expression of SV40 large cell antigen was detected in any of the cell lines. CONCLUSION: FACS analysis is more sensitive for evaluating mesothelin expression than immunohistochemistry of cut specimens. Irrespective of the expression of EGFR on FACS analysis, no EGFR mutation was detected. These three cell lines may be useful for studying cellular, molecular and genetic aspects of mesothelioma. PMID- 18383847 TI - Curcumin induces apoptosis in human neuroblastoma cells via inhibition of NFkappaB. AB - BACKGROUND: Metastasised neuroblastoma is a largely incurable neoplasia in children over one year of age using current treatment protocols. After dissemination to the bone, the survival rate is <7%, indicating an urgent need for novel therapeutic regimes. As curcumin (diferuloylmethane) had been shown to exert strong anticancer effects against diverse human malignancies different from neurblastoma, the antiproliferative effect of curcumin on the growth of human neuroblastoma cell lines was tested. MATERIALS AND METHODS: Proliferation of neuroblastoma cell lines Lan-5, SK-N-SH and Kelly under the treatment of curcumin over a broad concentration range (1 x 10(-5) to 1 x 10(2) microM) was assessed using XTT cell proliferation assays. Possible induction of apoptosis through curcumin treatment was assessed by detection of DNA fragmentation. To investigate the effect of curcumin on NFkappaB activation, the protein levels of the NFkappaB subunit p65 of curcumin-treated cells were compared to untreated cells using Western blots. RESULTS: Curcumin showed a significant dose-dependent antiproliferative effect on all three neuroblastoma cell lines starting at a concentration of 1 x 10(-3) microM. The highest concentration of 1 x 10(2) microM significantly reduced the viable cell count to 8-48% depending on the cell line. This antiproliferative effect was mediated through an increased induction of apoptosis by inhibition of NFkappaB, corroborating earlier findings indicating an antiapoptotic effect of NFkappaB. CONCLUSION: Our results suggest that curcumin might hold promise in the treatment of patients suffering from neuroblastoma. PMID- 18383848 TI - Frequent splicing aberration of the base excision repair gene hMYH in human gastric cancer. AB - BACKGROUND: Missense mutation of hMYH, which prevents transversion mutations induced by oxidative DNA damage, is reportedly associated with the development of gastric and colon cancer. We investigated whether deficiency or mutation of hMYH is associated with gastric carcinogenesis. PATIENTS AND METHODS: Thirty patients with gastric carcinoma, three gastric cancer cell lines and lymphocytes from three healthy volunteers were investigated. Reverse transcription-polymerase chain reaction (RT-PCR) was performed for hMYH, and the full-length sequence of hMYH mRNA was analysed. RESULTS: A silent mutation at codon 473 was seen in two tumours. Single nucleotide polymorphism at codon 345 was observed in 14 patients. These two base substitutions had no pathogenic effect. Seven splice variants were observed and two aberrant transcripts were detected more frequently in cancer specimens (67%) than in normal mucosa (10%). CONCLUSION: The high frequency of splicing aberration in cancer tissues suggests that aberrant transcripts may be involved in gastric carcinogenesis and cancer development. PMID- 18383849 TI - PAI-1, t-PA and circulating hTERT DNA as related to virus infection in liver carcinogenesis. AB - BACKGROUND: Liver carcinogenesis seems to be heavely influenced by hepatitis B and C viral (HBV, HCV) infection. The aim of our study was to improve the detection of hepatocellular carcinoma (HCC) by measuring alfa-fetoprotein (AFP) in addition to other molecular markers by estimating the plasma levels of human catalytic fraction of reverse telomerase (hTERT) DNA, plasminogen activator inhibitor-1 (PAI-1) and tissue plasminogen activator (t-PA) in 75 patients with liver desease. PATIENTS AND METHODS: A control group was enrolled (N=30). PAI-1 and t-PA levels were detected with enzyme-linked immunoassorbent assay (ELISA), DNA hTERT was performed with real time polymerase chain reaction (RT-PCR). RESULTS: PAI-1, t-PA and hTERT DNA levels were much higher than in controls. PAI 1 and t-PA levels were higher in the presence of both viruses compared to their absence, p<0.001. Moreover, hTERT was significantly higher in the presence of both viruses, p<0.05 and in the presence of HCV alone, p<0.05. No decrease or increase of AFP was noted in these patients. CONCLUSION: Our data suggest the reliability of PAI-1, t-PA and hTERT in detecting HCC, in particular when the carcinogenesis is affected by virus infection. PMID- 18383850 TI - Tumor-specific cytotoxicity and type of cell death induced by beta-cyclodextrin benzaldehyde inclusion compound. AB - The cytotoxicity of beta-cyclodextrin benzaldehyde inclusion compound (CDBA) against human normal and cancer cell lines was investigated. CDBA showed slightly higher cytotoxicity against human tumor cell lines, as compared to normal cells, with a tumor-specificity index of 2.2. Human myelogenous leukemia cell lines (HL 60, ML-1, KG-1) were the most sensitive to CDBA, followed by human oral squamous cell carcinoma (HSC-2, HSC-3, HSC-4) and human glioblastoma (T98G, U87MG). Human normal cells (gingival fibroblasts, pulp cells, periodontal ligament fibroblasts) were the most resistant. CDBA induced internucleosomal DNA fragmentation in HL-60 cells and caspase-3, -8, -9 activation, but to a much lesser extent than that attained by UV irradiation or actinomycin D. On the other hand, CDBA did not induce DNA fragmentation, nor caspase activation in HSC-2, HSC-4 or T98G cells. Electron microscopy demonstrated that CDBA induced the destruction of mitochondrial structure and digestion of broken organelles by secondary lysosomes in all of these cells. CDBA also increased the number of acidic organelles as judged by acridine orange staining. The present study suggests that CDBA induces autophagic cell death in cancer cell lines. PMID- 18383851 TI - Keratin profiling in the developing human prostate. A different approach to understanding epithelial lineage. AB - BACKGROUND: Keratin profiling studies in the developing human prostate have characterized cells thought to be stem cells and so-called intermediate cells. In a series of human prostates of various gestational ages, we extended on these studies using a comprehensive panel of keratin antibodies. MATERIALS AND METHODS: Autoptic tissue from 19 fetal prostates, gestational ages between 16 and 40 weeks, were immunostained with a panel of keratin antibodies: these recognize the luminal type keratins 7, 8, 18, 19, 20 and the basal/squamous type keratins 5, 6, 13, 14, 17. RESULTS: Keratin8 and vimentin were important constituents of the cytoskeleton of budding tips in early gestational age fetuses. Very early in gestation, additional expression of keratins 5 and 13 was noted and, with time, increasing expression of keratins 7, 18 and 19, and also incidentally keratins 14 and 17. With differentiation into basal cell and luminal cell compartments and the formation of prostate acini, the keratin complement of basal and luminal cells became more pronounced, but only partial compartmentalization of keratin expression occurred. CONCLUSION: We suggest that prostate stem cells may contain only keratin8 and not 5 or 14. The acquisition of other keratins could be indicative of the function the cells will eventually acquire. PMID- 18383852 TI - Stage-related decorin and versican expression in human laryngeal cancer. AB - BACKGROUND: The major proteoglycan of normal human larynx is aggrecan. In laryngeal carcinoma, aggrecan is depleted, with versican and decorin appearing in higher amounts. MATERIALS AND METHODS: Proteoglycans in laryngeal carcinoma samples were characterized immunohistochemically and using Western blotting; their expression was examined by RT-PCR. RESULTS: Aggrecan was totally removed in advanced cancer and its RT-PCR product was not identified. Both versican and decorin were overexpressed in cancer, versican much more than decorin. Decorin expression was higher than that of versican in the normal larynx; therefore, their disproportionate overexpression during cancer resulted in about equimolar expression. Both proteoglycans' expression correlated with their stage-related accumulation within the tissue. CONCLUSION: These data add to our previous findings and support the view that the levels of expression and the extent of accumulation and localization in the tumor stroma of structurally modified versican and decorin could be associated with the degree of aggressiveness of laryngeal carcinoma. PMID- 18383853 TI - Comparative analysis of cell death induction by cisplatin and 5-FU in human oral squamous and hepatocellular carcinoma cell lines. AB - We established the optimal conditions for the induction of cell death by cisplatin (CDDP) and 5-fluorouracil (5-FU) in human oral squamous cell carcinoma (HSC-2, HSC-3, HSC-4) and human hepatocellular carcinoma (HepG2) cell lines. HSC 3 cells were the most sensitive to 48 hours' continuous treatment with CDDP, followed by HepG2, HSC-2 and HSC-4 cells. On the other hand, HSC-4 cells were the most sensitive to 48-hour continuous treatment with 5-FU, followed by HSC-2, HSC 3 and HepG2 cells. CDDP induced internucleosomal DNA fragmentation in HSC-2 and HSC-3 cells, but not in HSC-4 cells, while 5-FU failed to induce internucleosomal DNA fragmentation in all of these cells. The treatment of HSC-2, HSC-3 and HSC 4cells with CDDP for 12 hours (followed by incubation for 36 hours without CDDP) showed comparable magnitude of cytotoxicity and caspase-3 activation with that attained by continuous 48-hour CDDP treatment. On the other hand, the cytotoxicity of 5-FU depended both on the dose and the exposure time. The present study demonstrate that the most effective treatment time is 12 hours for CDDP and much longer for 5-FU in all studied cell lines, underlining the importance of optimizing the treatment time for each chemotherapeutic agent. PMID- 18383854 TI - Adhesion to fibronectin induces megakaryocytic differentiation of JAS-REN cells. AB - BACKGROUND: Binding of integrins to the extracellular matrix elicits various responses. We have previously reported a megakaryocytic-erythroid cell line (JAS R) that showed phenotypic changes after adhesion to plastic dishes. However, the matrix protein and the mechanism responsible for megakaryocytic differentiation still remain unknown. MATERIALS AND METHODS: JAS-REN (erythroid) cells were cultured on dishes coated with various proteins. The cells were treated with RGDS, a tetrapeptide ligand to integrins, or phorbol ester (12-o tetradecanoylphorbol-13-acetate, TPA) for 48 hours and then were harvested. Subsequently, the cell surface markers were analyzed using flow cytometry and gene expression was studied by RT-PCR. RESULTS: The JAS-REN cells adhered to fibronectin-coated dishes, but showed poor adhesion to dishes coated with collagen, laminin or poly-D-lysine. The TPA-stimulated JAS-REN cells showed an increase in the expression of integrin alphaIIbbeta3 complex (CD41a) and integrin beta3 (CD61), while glycophorin A (CD235a) expression was decreased. JAS-REN cells that were adherent to fibronectin-coated dishes also showed a similar pattern of phenotype to TPA-treated cells, but the changes were not so prominent. RT-PCR revealed that TPA treatment altered the gene expression profile of JAS-REN cells, making it similar to that of JAS-RAD (megakaryocytic) cells. The RGDS treated and fibronectin adherent JAS-REN cells also showed a mostly similar expression profile to JAS-RAD cells, but these two stimuli did not alter the gene expression profile as TPA stimulation did. Transcription factors, FLI1 and GFI1, were induced by all stimuli. CONCLUSION: Signals triggered by adhesion to fibronectin result in the induction of FLI1 that may play a pivotal role in the lineage shift of JAS-REN cells from erythroid to megakaryocytic. PMID- 18383855 TI - A new single nucleotide polymorphism in XRCC4 gene is associated with breast cancer susceptibility in Taiwanese patients. AB - BACKGROUND: The DNA repair gene XRCC4, an important caretaker of the overall genome stability, is thought to play a major role in the human carcinogenesis. Some new and important polymorphic variants of XRCC4, at codon 247 (rs 3734091), G-1394T (rs 6869366), and Intron 7 (rs 28360317), and their association with breast cancer susceptibility was investigated in a Taiwanese population. MATERIALS AND METHODS: In a hospital-based case-control study, 432 female patients with breast cancer and 432 age-matched healthy controls recruited from the China Medical Hospital in Central Taiwan were genotyped. RESULTS: A significant difference in the frequency of the XRCC4 G-1394T genotype, but not the XRCC4 codon 247, or intron 7 genotypes was found between the breast cancer and control groups. Individuals with G/T or T/T at the XRCC4 G-1394T locus showed a 2.33-fold (95% confidence interval=1.37-3.98) increased risk of breast cancer compared to those with G/G. For XRCC4 codon 247 or intron 7, there was no difference in distribution between the breast cancer and control groups. CONCLUSION: Our findings suggest that the heterozygous and homozygous T allele of the XRCC4 G-1394T may be associated with the development of breast cancer and may be a useful biomarker for anticancer prevention and intervention. PMID- 18383856 TI - A DNA polymorphism of stromal-derived factor-1 is associated with advanced stages of oral cancer. AB - BACKGROUND: Stromal derived factor-1 (SDF-1), a protein related to angiogenesis and inflammation, has been correlated with the progression of a number of malignancies, but not with oral squamous cell carcinoma. In light of the known contribution to the development of oral cancer of other gene polymorphisms for proteins responsible for angiogenesis, inflammation and thrombosis, this study investigated whether the G801A polymorphism in the SDF-1 gene is associated with this malignancy. PATIENTS AND METHODS: The G801A polymorphism was examined by restriction fragment length polymorphism analysis in DNA samples from 159 patients with oral squamous cell carcinoma and 101 matched healthy controls. RESULTS: The detected allele frequency of the "high production related" A allele in the control group was 25.3%. There was a slight decrease in allele frequency in patients (18.6%), but it was not statistically significant. The same pattern was observed in subgroups of patients in regard to smoking habits and family history of cancer or thrombosis. Interestingly, in comparison to controls, the A allele frequency was significantly lower in patients with advanced cancer stages III and IV (12.5%, p=0.005) and in patients with alcohol abuse (12.5%, p=0.02). CONCLUSION: The G801A polymorphism of the SDF-1 gene is associated with advanced stages of oral cancer, especially in alcohol abusers. PMID- 18383857 TI - Quantitative structure-cytotoxicity relationship analysis of 3-formylchromone derivatives by a semiempirical molecular-orbital method with the concept of absolute hardness. AB - A semiempirical molecular-orbital method (CAChe) was applied to delineate the relationship between the cytotoxicity (evaluated by 50% cytotoxic concentration, CC50) of 11 3-formylchromone derivatives and 15 chemical parameters (descriptors). The most stable conformation of all these compounds was exhibited by the planar structure. Compounds [2], [3], [4] and [9] had additionally protruding branches from the coplanar. In HSG cells, the best correlation coefficient was observed between CC50 and stability of hydration (DeltaH), followed by electron affinity, lowest unoccupied molecular orbital energy (ELUMO), highest occupied molecular orbital energy (EHOMO), ionization potential, absolute electron negativity (chi) and reactivity index (omega). When the value for [1], which was off from the regression line, was omitted, higher correlation coefficients were obtained between CC50 and electron affinity, ELUMO, chi and omega. When CC50 value was plotted vs. log P, a parabolic curve was produced, under the condition that the data for [5] were omitted. In HL-60 cells, moderate correlation was found between CC50 and DeltaH, electron affinity, ELUMO, chi and omega. When the values for [1] and [6], which were off the regression line, were omitted, higher correlation coefficients were obtained between CC50 and these five descriptors. In HSC-3 cells, there was moderate correlation between CC50 and the dipole moment, but not with other descriptors. In HSC-2 and MT-4 cells, there was no clear-cut correlation between CC50 and any of these descriptors. The present study indicates the applicability of HSG cells in searching for more active 3-formylchromone derivatives, using QSAR with the concept of absolute hardness. PMID- 18383858 TI - Growth inhibition and proapoptotic activity induction by IIF and valproic acid on RA-resistant leukemia cells. AB - All-trans retinoic acid (RA) induces complete remission in a high proportion of patients with acute promyelocytic leukemia (APL). Nevertheless, most of these patients develop RA resistance and relapse. In an attemp to mimic clinical conditions for the treatment of leukemia, a stably RA-resistant subclone of the human promyelocytic leukemia cell line HL60 (HL60-R) was developed to study the antiproliferative and proapoptotic effect of the retinoid IIF (6-OH-11-O hydroxyphenantrene) in comparison with RA. Moreover whether the inhibitor of histone deacetylase (HDAC) activity, valproic acid (VPA), could enhance sensitivity to retinoids in HL60-R cells was evaluated. Finally, the effect of IIF on the expression of multidrug resistance-associated protein 1 (MRP1) and P glycoprotein (P-gp) was evaluated. It was found that IIF strongly suppressed cell proliferation (as measured by growth curves) and induced apoptosis (as measured by DNA fragmentation and Annexin V detection assays), while RA was practically ineffective. The addition of VPA to IIF accentuated the antiproliferative effect of IIF alone and increased apoptosis; the combination of VPA with RA allowed growth arrest. Moreover IIF caused a reduction of transmembrane transporter expression, particularly of P-gp, as shown by Western blotting. Our results suggest that IIF may be useful in controlling the proliferation of RA-resistant leukemia cells, especially in combination with an HDAC inhibitor, such as VPA. PMID- 18383859 TI - Antiangiogenic properties of lupulone, a bitter acid of hop cones. AB - BACKGROUND: Angiogenesis is the result of intricate steps regulated by the balance between agonistic and antagonistic effectors. Disturbance of this balance leads to an 'angiogenic' switch critical for tumor development. MATERIALS AND METHODS: Using human umbilical vein endothelial cells (HUVEC) the effects of lupulone were analyzed on proliferation induced by angiogenic growth factors, transmembrane cell migration toward fibronectin and formation of a network of tubular-like structures on Matrigel. RESULTS: Lupulone (2.5-50 microg/ml) induced a concentration-dependent inhibition of HUVEC proliferation and chemotaxis. Lupulone caused a significant reduction of closed capillary-like structures in Matrigel indicating a strong inhibitory effect on neovascularisation. In mice receiving lupulone (20 mg/kg/day) in drinking water for 21 days, new vessel formation was reduced by 50% in matrigel plugs implanted under the skin when compared with controls. CONCLUSION: The present data demonstrate that lupulone is able to inhibit angiogenesis in vitro and in vivo. Lupulone emerges as a potential chemopreventive agent when considering its strong antiangiogenic properties. PMID- 18383860 TI - Immunohistochemical expression patterns of neural and neuroendocrine markers, the neural growth factor receptors and the beta-tubulin II and IV isotypes in human thymus. AB - Increasing evidence suggests that neuroimmune networks play key roles in the thymic histophysiology and pathology. Prompted by this, we analyzed by immunohistochemistry the distribution of human thymic cells expressing major neural and neuroendocrine markers and neural growth factor (NGF) receptors in combination with the expression patterns of various cytokeratins. Additionally, since some beta-tubulin isotypes are preferentially expressed in neuronal cells, the immunotopographical distribution of thymic cells expressing beta-tubulin II, III and IV was analyzed. Thymic epithelial cells (TECs) expressed protein gene product 9.5 (PGP 9.5), chromogranin A (CHRA), synaptophysin (SYN), neuron specific enolase (NSE), tyrosine hydroxylase (TH), CD56, CD57, neurofilaments (NF) (140-160 kDa), NGF receptors (TrKA and p75), beta-tubulin II and IV isotypes and cytokeratin 7, 8, 10, 13, 14, 18 and 19. PGP 9.5 was preferentially expressed in cortical TEC whereas SYN, CHRA, NSE, TH and NF 140-160 kDa were preferentially expressed in medullary TECs and Hassal corpuscles. Variable levels of expression of beta-tubulin II and IV were observed in all TEC subtypes whereas beta-tubulin III was undetectable in TECs. Subcapsular and cortical TECs display higher expression of beta-tubulin IV and lower expression of beta-tubulin II in comparison to those observed in medullary TEC and Hassal corpuscles. The diversity of the immunotopographical distibution and the expression of neural and neuroendocrine markers, the NGF receptors TrKA and p75, and the beta-tubulin II and IV isotypes in the distinct subtypes of TEC may reflect the diversity of their biological functions and/or their different stages of differentiation. The present results provide further immunohistological evidence that numerous neural and neuroendocrine factors may be required for the development and function of the human thymic microenvironment. PMID- 18383861 TI - BRAF and K-RAS mutation in a Greek papillary and medullary thyroid carcinoma cohort. AB - BACKGROUND: The genes RAS and BRAF have been shown to be frequently mutated in human thyroid carcinomas. The aim of this study was to genotype a cohort of 55 sporadic papillary thyroid carcinomas (PTC) and 44 sporadic medullary thyroid carcinomas (MTC) for the K-RAS codon 12 and BRAF codon 600 mutations. MATERIALS AND METHODS: K-RAS and BRAF mutations were characterized by an enhanced polymerase chain reaction followed by restriction fragment length polymorphism analysis (PCR-RFLP). RESULTS: The K-RAS codon 12 mutation was found in 54.5% of the PTC and 40.9% of the MTC cases tested. The BRAF V600E mutation was detected in 27.3% of the PTC and 68.2% of the MTC samples. No significant association between K-RAS and BRAF mutations and clinicopathological parameters was found. CONCLUSION: These data indicate that K-RAS and BRAF mutations were a frequent genetic event in our samples of sporadic PTC and MTC. PMID- 18383862 TI - The interleukin-10 (-1082A/G) polymorphism is strongly associated with increased risk for oral squamous cell carcinoma. AB - BACKGROUND: Increased levels of interleukin-10 (IL-10) have been observed in patients with oral cancer, possibly as a result of suppression of the immune response. Based on this, the -1082A/G polymorphism, which influences IL-10 gene expression level, was investigated in regard to its possible association with risk for oral cancer. PATIENTS AND METHODS: The polymorphism was examined in DNA samples of 144 patients with oral squamous cell carcinoma and 141 healthy controls of equivalent gender, age and ethnicity. RESULTS: The detected allele frequencies for the high expression G allele were significantly higher in patients compared to controls (34.7% versus 21.3%, respectively, p=0.0004), as well as in patients that were smokers but not those that were heavy alcohol consumers. This highly significant difference in G allele frequency was mainly due to the increase of AG heterozygotes in patients compared to controls (OR 3.05, 95% CI 1.84-5.05). CONCLUSION: These findings suggest that the high expression G allele of the -1082A/G polymorphism of the inflammation and angiogenesis-related IL-10 is strongly associated with increased risk for oral cancer. PMID- 18383863 TI - Pro-apoptotic activity of cyclopentenone in cancer cells. AB - Studies on cyclopentenone prostaglandins (CPPGs), clavulones and other cyclopentenones have shown that these compounds have a significant anticancer activity mediated by their cyclopentenone (CP) chemical moiety. In this study the cytotoxicity against cancer cells of the model compound cyclopent-2-en-1-one (2CP) was investigated. Being a highly water soluble small molecule, 2CP could be an ideal candidate to overcome pharmacological issues related to drug delivery and penetration. Its cytotoxic activity was tested on various melanoma and lung cancer cells. Interestingly, 2CP was both cytotoxic and pro-apoptotic, more pronounced on melanoma cells, at concentrations in the sub-micromolar range. On melanoma cells its mechanism of action was mediated by the mitochondria and the activation of caspase 3. PMID- 18383864 TI - Elevated serum-circulating RNA in patients with conventional renal cell cancer. AB - BACKGROUND: Reliable serum biomarkers for differential diagnosis of conventional renal cell carcinoma (RCC) are highly desirable. Recent studies have confirmed the stability of circulating RNA in serum of cancer patients. The purpose of our study was to evaluate whether the amounts of circulating RNA could discriminate between conventional renal cancer patients and healthy individuals as a tumor marker. PATIENTS AND METHODS: A total of 71 patients with conventional RCC, 12 with renal oncocytomas and 44 healthy individuals entered into this study. Serum samples were taken and subjected to RNA extraction. The amount of RNA was quantified spectrophotometrically. Additionally, 9 serum samples from conventional RCC were also studied one week after nephrectomy. Diagnostic performance of RNA concentration was calculated through the receiver operating characteristic (ROC) curve to distinguish between conventional RCC and healthy individuals. RESULTS: The mean level of RNA in conventional RCC (1414.19 +/- 91.95 ng/ml) was significantly higher than that in healthy individuals (520.49 +/ 39.75 ng/ml, p<0.0001) and these with renal oncocytomas (560.71 +/- 69.54 ng/ml, p<0.0001). Among the conventional RCC, there was no significant difference in circulating RNA levels in terms of tumor stage, grade or size. The area under the ROC curve was 0.956 (95% confidence interval, 0.923 to 0.989), indicating an acceptable sensitivity and specificity as a tumor marker. For conventional RCC, the RNA level was reduced significantly (p<0.0001) one week after nephrectomy. CONCLUSION: The data suggest that elevated circulating RNA may be a valuable diagnostic tool for discriminating conventional RCC patients from normal individuals or from these with renal oncocytoma. Elevated serum circulating RNA provides a new research area as biomarker for the diagnosis of conventional RCC. PMID- 18383865 TI - Overexpression of vimentin contributes to prostate cancer invasion and metastasis via src regulation. AB - A significant proportion of prostate cancer patients treated with curative intent develop advanced disease. At a fundamental biological level, very little is known about what makes the disease aggressive and metastatic. Observational pathology reports and experimental data suggest that an epithelial-mesenchymal transition (EMT) is involved in prostate cancer invasiveness. The mechanism by which vimentin promotes prostate cancer cell invasion and metastasis was examined. The highly metastatic human prostate epithelial cell line PC-3M-1E8 (1E8-H) and the low metastatic line PC-3M-2B4 (2B4-L) were used for comparative proteomic analysis by two-dimensional gel electrophoresis, followed by matrix-assisted laser desorption/time of flight mass spectrometry (MALDI-TOF-MS). A transwell assay was performed to test cell migration and invasion and immunoblotting was used to analyze the relative expression of proteins. High vimentin expression was detected in 1E8-H compared to 2B4-L cells. Down-regulation of vimentin in 1E8-H by antisense-vimentin transfection led to a significant inhibition of invasiveness, and selective stimulation of vimentin activity in 2B4-L by delivery of recombinant vimentin promoted cell invasiveness. Vimentin activity was associated with C-src, beta-catenin and E-cadherin expression. PP2, a specific inhibitor of src family kinases, reduced phospho-beta-catenin expression and induce E-cadherin expression. Vimentin promotes tumor cell invasiveness and the targeting of vimentin/C-src may be a promising strategy for preventing or blocking prostate cancer metastasis. PMID- 18383866 TI - Valproate protective effects on cisplatin-induced peripheral neuropathy: an in vitro and in vivo study. AB - BACKGROUND: Antineoplastic drugs, such as cisplatin (CDDP), induce disabling peripheral neuropathies, representing a hindrance to effective cancer treatments. The exact pathogenesis of CDDP-induced neuropathy is not yet understood, and the dysregulation of gene expression has been proposed. Valproate (VPA) is an antiepileptic drug recently discovered to remodel gene expression, with hypothetically putative neuroprotective effects. MATERIALS AND METHODS: VPA was tested in both, in vitro and in vivo models of CDDP-neurotoxicity. RESULTS: VPA administered in combination with CDDP promoted dorsal root ganglia (DRG) neurons survival. Moreover, this treatment induced in Wistar rats an improvement of body weight, sensory nerve conduction velocity, and DRG morphometric analysis. In contrast, VPA was not able to rescue CDDP pre-treated rats. CONCLUSION: When used in combination with CDDP, VPA displays a protective action against neuropathy, in our models, suggesting possible future clinical applications. PMID- 18383867 TI - Prognostic impact of lymphangiogenesis and lymphatic invasion (CD105 expression) in small cell lung carcinoma. AB - BACKGROUND: Lymphangiogenesis, an essential process in the metastasis of malignant tumors, has not been thoroughly studied. The possibility of using it to define subsets of patients with different prognosis in cancer could be of vital clinical importance. MATERIALS AND METHODS: Fifty patients (5 women, 45 men; mean age, 64.47 years) with SCLC were retrospectively studied. Tumor specimens were stained for CD105, and intratumoral lymphatic microvessel density (ILMVD) and lymphatic invasion were determined. RESULTS: Twenty-five patients were diagnosed with limited and 25 with extensive SCLC. All patients received chemotherapy and 32.7% radiation therapy. A direct association between ILMVD (CD105 expression) and lymphatic invasion was observed (p<0.046). CD105 expression was significantly associated with the stage of the disease (p=0.004) and the presence of metastasis (p=0.05). CONCLUSION: CD105 expression and lymphatic invasion correlated significantly with the clinical parameters and patient outcome, therefore, constituting an important prognostic role in SCLC. PMID- 18383868 TI - Lack of genotoxicity of carbon nanotubes in a pilot study. AB - BACKGROUND: Different types of carbon nanotubes may represent toxic hazards due to their size distribution and massive surface area. They may adsorb other toxic agents that can consequently be transported into the body. The aim of the present study was to determine the possible genotoxicity of carbon nanotubes. MATERIALS AND METHODS: In vivo bacterial mutagenicity and in vitro cytogenetic studies were performed on single-walled and multi-walled carbon nanotubes. RESULTS: Oral exposure to nanotubes did not increase urinary mutagenicity in rats as studied using Ames test. No genotoxic effect was found in the in vitro micronucleus and sister chromatid exchange assays, either. Mitotic inhibition, a possible cytotoxic effect, however, was observed in the human lymphocyte cultures upon treatment with single-walled tubes. CONCLUSION: Due to the limited toxicity data on carbon nanotubes, these results may be particularly important for risk assessment purposes. PMID- 18383869 TI - Confocal endomicroscopy: in vivo diagnosis of neoplastic lesions of the gastrointestinal tract. AB - Confocal endomicroscopy is a novel technology which allows subsurface histological diagnosis at a cellular and subcellular level in vivo. It thereby provides instantaneous histopathology during ongoing upper and lower endoscopy. This allows immediate diagnosis of neoplastic and inflammatory lesions of the intestinal mucosa. Studies have demonstrated the power of confocal endomicroscopy in screening and surveillance colonoscopy, ulcerative colitis, Barrett's esophagus, and gastric cancer. In animal models of human diseases, the same technology has provided molecular imaging of cancer, functional imaging of altered perfusion in malignant and inflammatory disease and high resolution in vivo morphological diagnosis. Fields of ongoing research are the development of molecular markers for in vivo immunohistochemistry and the application of confocal microscopy to intraabdominal organs in humans. Confocal endomicroscopy is evolving as a novel technique for rapid intravital diagnosis of gastrointestinal neoplastic diseases at the microscopic level and bears the potential for molecular imaging in humans in the future. PMID- 18383870 TI - Pancreatic resection for metastatic renal cell carcinoma: a single center experience and review of the literature. AB - Renal cell carcinoma (RCC) is one of the few tumour types metastatic to the pancreas. In order to evaluate the outcome following resection of pancreatic metastases of RCC a retrospective review of surgical patients was performed. The initial histopathological staging, disease-free interval, surgical outcome and survival were evaluated. The median interval between nephrectomy and pancreatic resection was 9 years. Six out of the ten patients preoperatively presented with severe complaints caused by the pancreatic metastasis, such as pain, chronic pancreatitis, jaundice and gastrointestinal bleeding. Severe postoperative complications only occurred in two patients, who presented in a deteriorated condition preoperatively. The median follow-up was 56 months, in 3 patients more than 5 years. Although the spontaneous course of RCC metastases can be favourable, the complete resection of pancreatic metastases for patients in good physical condition is suggested if possible. Moreover, good palliation of symptoms in patients with long-term survival can be achieved. PMID- 18383871 TI - Comparative analysis of four histopathological classification systems to discriminate benign and malignant behaviour in gastrointestinal stromal tumors. AB - The prognostic value of the four most common histopathological classification systems in gastrointestinal stromal tumors (GISTs) was evaluated retrospectively. PATIENTS AND METHODS: Twenty-five consecutive patients with resected GIST and a follow-up of five years or more for surviving patients were included in this analysis. All the tumors were c-KIT (CD117) positive and were additionally re evaluated for the number of mitoses per 50 high-power fields (HPF) and Ki-67 proliferation index. The four most commonly applied histopathological classification systems of the WHO, Franquemont (modified by using the Ki-67 proliferation index), Fletcher and Miettinen were applied to each patient. RESULTS: The survival of patient groups classified by Franquemont (p = 0.03) and the WHO (p = 0.031) were of prognostic relevance, while the grouping of patients by classifications according to both, Fletcher and Miettinen did not show a significant prognostic value. CONCLUSION: The classification systems of Franquemont (modified) or WHO appear to be advantageous for the evaluation of malignant potential and clinical outcome in patients with GISTs. Our data are merely hypothesis generating and should be validated in larger clinical studies. PMID- 18383872 TI - Changes of immunological parameters reflect quality of life-related toxicity during chemotherapy in patients with advanced colorectal cancer. AB - Anticancer drugs may frequently induce host immunosuppression and symptomatic toxicities. Once symptomatic toxicity occurs, the patient's quality-of-life (QOL) is reduced. Since little is known of the relationship between host immunity and the toxicity of chemotherapy, the host immunity before and after chemotherapy was compared to assess whether it is related to symptomatic toxicity during chemotherapy. PATIENTS AND METHODS: Fourteen patients with colorectal cancer underwent leucovorin /5-fluorouracil (LV/5-FU) treatment, or S-1/irinotecan (CPT 11). Host immunity (cytokine production of peripheral blood mononuclear cell (PBMC), serum soluble interleukin-2 receptor (sIL-2R) levels and phenotypic analyses of PBMC were measured before and after the first chemotherapy. RESULTS: An increase of sIL-2R, CD4+CD25+ T-cells and the CD4/8 ratio in patients with symptomatic adverse reactions were found. These changes in the first chemotherapy were significantly different (p = 0.0211, p = 0.0087, p = 0.0234). CONCLUSION: The current study indicated that there are some parameters correlated with toxicity during chemotherapy which effect QOL. In such patients, negative influences on host immunity, such as an increase of sIL-2R and regulatory T cells, and a decrease of cytotoxic T-cells could occur. PMID- 18383873 TI - Adoptive immunotherapy for pancreatic cancer using MUC1 peptide-pulsed dendritic cells and activated T lymphocytes. AB - BACKGROUND: Pancreatic cancer has a poor prognosis. The clinical efficacy of immunotherapy using both dendritic cells pulsed with MUC1 peptide (MUC1-DC) and, cytotoxic T lymphocyte (CTL) sensitized with a pancreatic cancer, YPK-1, expressing MUC1 (MUC1-CTL) was evaluated. PATIENTS AND METHODS: Twenty patients with unresectable or recurrent pancreatic cancer were enrolled. Peripheral blood mononuclear cells (PBMCs) were separated into adherent cells for induction of MUC1-DCs and floating cells for MUC1-CTLs. MUC1-DCs were generated by culture with granulocyte monocyte colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) and then exposed to MUC1 peptide and TNF-alpha. MUC1-CTLs were induced by co-culture with YPK-1 and then with interleukin-2 (IL-2). MUC1-DCs were injected intradermally and MUC1-CTLs were given intravenously. RESULTS: Patients were treated from 2 to 15 times. One patient with multiple lung metastases experienced a complete response. Five patients had stable disease. The mean survival time was 9.8 months. No grade II-IV toxicity was observed. CONCLUSION: Adoptive immunotherapy with MUC1-DC and MUC1-CTL may be feasible and effective for pancreatic cancer. PMID- 18383874 TI - 5-Fluorouracil-related gene expression in hepatic artery infusion-treated patients with hepatic metastases from colorectal carcinomas. AB - AIM: To predict the therapeutic efficacy of hepatic arterial infusion (HAI) with 5-fluorouracil (5FU) for patients with liver metastases from colorectal carcinomas, 5FU-related gene expressions were examined in primary colorectal carcinomas. PATIENTS AND METHODS: Thirty-eight patients with liver metastases from colorectal carcinoma received HAI of 5FU. The expressions of the mRNAs for thymidine synthase (TS), dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP), and oroteta phophoribosyl transferase (OPRT) in primary colorectal carcinomas were measured by RT-PCR. RESULTS: The response rate was 52.6% (20/38). The overall median survival time was 29.1 months. DPD and TP expression was significantly higher in the progressive disease (PD) group than in the complete response (CR) or partial response (PR) group (p = 0.032, p = 0.014), respectively. The levels of DPD and TP mRNAs showed a significant correlation (r = 0.76, p = 0.0001). CONCLUSION: The expression of DPD and TP mRNAs in primary colorectal carcinomas was significantly predictive of the therapeutic response to 5FU HAI. PMID- 18383875 TI - Alpha 1 antichymotrypsin genotype is associated with increased risk of prostate carcinoma and PSA levels. AB - BACKGROUND: Prostate cancer (PCa) is the most common non-skin cancer among men in Western countries. Inflammation appears to be involved in the pathogenesis of PCa. Recent studies have shown that many inflammatory genes are associated with the risk of PCa. Alpha 1 antichymotrypsin (ACT) is an acute phase protein and it is part of the circulating prostate specific antigen (PSA). PATIENTS AND METHODS: Allele and genotype frequencies of a promoter single nucleotide polymorphism (SNP) in ACT gene were investigated in patients with benign prostate hypertrophy (BHP) or PCa and controls. RESULTS: The G allele was more represented in PCa patients (odds ratio = 2.349). The PSA levels and prostatic volume did not correlate with the ACT genotype. However, stratifying subjects by age, a correlation of PSA levels and the GG genotype in young PCa patients was found. CONCLUSION: Carriers of the ACT G allele are at risk of developing PCa and genotyping healthy subjects could be a new approach for early prevention. PMID- 18383876 TI - The ATM gene and ataxia telangiectasia. AB - Ataxia telangiectasia (AT) is a rare neurodegenerative, autosomal recessive disorder characterized by chromosome instability, radiosensitivity, immunodeficiency and a predisposition for cancer. Epidemiological studies have shown that AT heterozygotes have a predisposition for cancer, especially for breast cancer in women. The disease is caused by mutations in the ATM gene, leading to total loss of the ATM protein, which normally recognizes DNA damage, activates the DNA repair machinery and the cell cycle check points in order to minimize the risk of genetic damage. This review summarizes the clinical features of AT and the natural history of the disease and puts recent molecular advances into the context of the cellular and clinical phenotype. PMID- 18383878 TI - High serum level of plasminogen activator inhibitor-1 predicts histological grade of intracerebral gliomas. AB - BACKGROUND: No serum marker is currently available for the diagnosis and treatment of gliomas. Plasminogen activator inhibitor-1 (PAI-1) controls the proteolytic activity in cancer cells and cellular migration during angiogenesis. PATIENTS AND METHODS: To verify the potential of PAI-1 as a serum marker for gliomas, the serum PAI-1 concentrations were measured by ELISA in 57 glioma patients and 34 healthy volunteers. RESULTS: We found significantly higher serum levels in the patients with high-grade gliomas than in the healthy volunteers (p = 0.0009, unpaired t-test) and those with low-grade tumors (p = 0.0074). Furthermore, high-grade glioma patients with a low serum level of PAI-1 survived significantly longer than those with high levels (p = 0.0082). Immunohistochemical analysis using anti-PAI-1 antibody revealed dense and spotty staining in the high-grade tumor tissues from the patients with high serum PAI-1 levels. CONCLUSION: These results suggest that the serum PAI-1 level can be a marker for the prediction of histological grade in intracerebral glioma. PMID- 18383877 TI - Metastatic adenocarcinoma in the brain: magnetic resonance imaging with pathological correlations to mucin content. AB - BACKGROUND: Hypointense signal appearance of metastatic adenocarcinoma on T2 weighted imaging (T2-WI) has been infrequently documented. The purpose of this report was to evaluate the degree to which mucin content affects signal manifestations on conventional MR imaging. PATIENTS AND METHODS: This series of 24 cases with intracerebral metastatic adenocarcinoma was assessed retrospectively, focusing on the association between hypointense appearance on T2 WI and intratumoral mucin content. RESULTS: Among the 24 metastatic adenocarcinomas, intratumoral mucin was histopathologically confirmed in 8 lesions. Of these, 4 masses were demonstrated as hyperintense signal on T2-WI. The other 4 masses were depicted as isointensity. No cases were identified with hypointense signals in mucin-containing metastatic adenocarcinoma. Conversely, only 2 metastatic tumors originating from the stomach exhibited hypointense signal lesions on T2-WI. No histological or magnetic resonance imaging evidence of blood products or other forms of iron were identified as causes of the hypointense appearance on T2-WI. CONCLUSION: This hypointense signal may simply reflect the relatively shorter T2 relaxation time of the primary tissue from which metastases arose. Intratumoral mucin itself may be considered to demonstrate the masses as hyper- or isointense signals in the brain. PMID- 18383879 TI - Axillary and subcutaneous breast metastases from rhinopharyngeal carcinoma: a case report and literature review. AB - The incidence of rhinopharyngeal carcinoma is fewer than one person per 100,000 population but it is one of the most common types of cancer in Southern China. The mechanisms often implicated suggest an important role of genetic, ethnic and environmental factors. Lung metastases are the most frequent, accounting for 66% of distant metastases. Other metastatic sites include bone (22%) and liver (10%). We report a case with breast skin and axillary nodes involvement. PMID- 18383880 TI - Prognostic value of the androgen receptor and its coactivators in patients with D1 prostate cancer. AB - BACKGROUND: Prostate cancer treated with androgen ablation eventually becomes resistant. Because the androgen receptor (AR) signaling axis affects disease progression, AR coactivator molecules could provide clinical prognostic value. This study investigates the association between AR coactivator molecules and clinical outcome measures in patients with prostate cancer. PATIENTS AND METHODS: Expression levels of AR and its coactivators, SRC1, TIF2, and Her2/neu were determined by quantitative RT-PCR in 148 prostatectomy specimens. AR protein expression was determined by immunohistochemistry. The prognostic value of these expression levels on clinical outcomes was examined. RESULTS: Increased gene and protein AR expression was not correlated with any of the clinical outcome measures. A non-monotonic correlation was observed between SRC1 and overall survival, as well as Her2/neu and time to prostate-specific PSA recurrence. CONCLUSION: Although no statistically significant relationships were found, the weak association between some clinical outcomes and two AR coactivators may help improve the current predictive nomogram for patients with prostate cancer. PMID- 18383881 TI - Dysphagia severity and aspiration following postoperative radiation for locally advanced oropharyngeal cancer. AB - The aim of the present study was to assess dysphagia severity following postoperative radiation for locally advanced oropharyngeal cancer. PATIENTS AND METHODS: Eighteen patients with oropharyngeal carcinoma had undergone postoperative radiation. There were eight base of tongue, eight tonsils, and two soft palate carcinomas. All the patients had undergone modified barium swallow (MBS) to assess the persistence of dysphagia (more than one month) post treatment. All the patients were cancer-free at the time of the swallowing study. Dysphagia severity was graded as 1-7. RESULTS: At a median follow-up of 12 months, there were three grade 2, four grade 3, two grade 4, five grade 5, two grade 6, and two grade 7. Only three patients (17%) had normal swallow post treatment. Six patients (33%) had mild to moderate dysphagia (grade 3-4). Nine patients (50%) developed aspiration (grade 5-7). Among the patients who developed aspiration, four (22%) required tube feeding for severe aspiration. CONCLUSION: Long-term (more than one year) dysphagia following postoperative radiation for oropharyngeal cancer may be symptomatic of permanent damage to the swallowing mechanism. Evaluation of patients who complain of persistence of dysphagia a year or more following treatment should include MBS, because of the increased risk of aspiration. PMID- 18383882 TI - Lymph node metastasis density (ND)-factor association with malignant degree and ND40 as "non-curative factor" in gastric cancer. AB - Advanced gastric cancer is not controlled by surgery alone, although early gastric cancer can be cured by curative gastrectomy. The TNM factor, representing physical tumor spread, accurately predicts patient prognosis in advanced gastric cancer, however, more elaborate prognostic factors reflecting the malignant degree are necessary in order to determine the therapeutic target. The purpose of this study was to select a significant prognostic indicator of gastric cancer with curative intent, independent of TNM stage among daily-feasible clinical parameters. The current study included 382 patients with stage IB to III gastric cancer after curative surgery, in which the robust univariate prognostic predictors were age (p < 0.0001), lymph node metastasis density of 40% or over (ND40) (p < 0.0001), and preoperative high value of carcinoembryonic antigen (CEA) (preCEA) (p = 0.0012), as well as TNM stage, preoperative high value of CA19-9 (preCA19-9), a glyco-chain antigen recognized by monoclonal antibody NS19 9, and vascular invasion. In multivariate prognostic analysis, age (p = 0.0002), ND40 (p = 0.02), and preCEA (p = 0.03) remained independent prognostic factors. The ND-factor provided excellent prognostic stratification within the same stage, and stage III ND40 predicted a similar prognosis to stage IV and could be considered a non-curative factor. Interestingly, ND40 relevance was validated in the prospective and independent set of gastric cancer patients (p = 0.002). CONCLUSION: The ND factor reflects the malignant degree of gastric cancer, and ND40 is a novel "non-curative factor" in gastric cancer patients. A search for the molecules associated with higher ND value within an identical stage could lead to objective determination of the degree of malignancy and development of novel strategies against advanced gastric cancer. PMID- 18383883 TI - DNA image cytometry in bladder cancer: state of the art. AB - The most recent Consensus Review of the Clinical Utility of DNA Cytometry in Bladder Cancer, which took place in Maine back in 1992, focused solely on flow cytometry results. Since then, there have been a significant number of articles published on the use of image cytometry to evaluate DNA content in bladder cancer. This has meant that a large proportion of the information collected is scattered across the published literature. The purpose of this article was to organize the data referred to in those articles published since the 1992 Consensus Review, and organise it under three major topic headings: a) DNA image cytometry versus flow cytometry, (b) specimen sources, and (c) its clinical utility with regard to improving prognosis and recurrence detection. A variety of factors and issues are discussed and points have been raised for discussion. Prospects for the future and potential research areas are also suggested. PMID- 18383884 TI - MCM-2, Ki-67 and PCNA protein expressions in pT3G2 colorectal cancer indicated lymph node involvement. AB - BACKGROUND: The rate of cancer cell growth depends on proliferative activity and on the tumor cell death rate. The study objective was to assess the expressions of Ki-67, PCNA and MCM2 in colorectal cancer and to analyze the correlation between the expressions and chosen anatomo-clinical parameters (patient's age, tumor location, lymph node metastases). PATIENTS AND METHODS: The study material included primary tumors pT3 G2 obtained from 55 patients with colorectal cancer. Immunohistochemical investigations were performed using monoclonal antibodies (Ki 67- monoclonal mouse (DAKO) Clone MIB1, PCNA-monoclonal mouse (DAKO) Clone PC10, MCM2-polyclonal goat (Santa Cruz Biotechnology) Sc-9839) directed against human protein Ki-67, PCNA and MCM2. RESULTS: The expressions of Ki-67, PCNA and MCM2 were found to correlate with the presence of lymph node metastases, but not with patient age or tumor location. The expressions of these proteins in the main tumor mass correlated with each other in the majority of patients. CONCLUSION: The findings allow the assumption that positive expressions of Ki-67, PCNA and MCM2 in the main tumor mass in pathological stage pT3 and degree of histological malignancy G2 may indicate lymph node involvement. PMID- 18383885 TI - Practice and costs of red blood cell (RBC) transfusion in an oncological unit. AB - BACKGROUND: Cancer related anemia impairs patient functioning. Red blood cell (RBC) transfusion and erythropoietin (EPO) may relieve fatigue. Cost effectiveness data have been requested. PATIENTS AND METHODS: All transfusions administered at the Department of Oncology, University Hospital of North Norway (UNN) in 2005 were analysed, with a total of 118 patients entering the study. A cost of transfusion analysis was added and a sensitivity analysis conducted to clarify robustness. The 118 patients received 613 units of erythrocytes. In 6% of cases, the transfusion was the only cause of a hospital visit. One fourth of patients had bone marrow infiltration and two-thirds had undergone chemotherapy. The mean Hb levels of patients prior to and following transfusion were in the range 8.4-8.8 g/dl and 10.2-10.6 g/dl, respectively; one-third reached a non anemic level (Hb > or = 11.0 g/dl). The median time interval between transfusions was three weeks and the annual cost was calculated at Euro 1,069/patient. CONCLUSION: RBC-transfusion has a low cost. PMID- 18383886 TI - 94% accuracy of intraoperative imprint touch cytology of sentinel nodes in skin melanoma patients. AB - The aim of the present study was to assess whether the reliability of imprint touch cytology (ITC) of sentinel nodes in skin melanoma patients allows intraoperative decisions regarding simultaneous radical lymphadenectomy to be made. PATIENTS AND METHODS: The results of ITC of sentinel nodes were compared with the results of standard histopathological and immunohistochemical examinations. RESULTS: A total of 148 sentinel nodes were identified in 98 lymph node groups in 85 skin melanoma patients. ITC revealed the presence of metastases in 7 out of 16 melanoma-positive sentinel nodes (sensitivity, 43.7%). There were no false-positive results of ITC of sentinel nodes (specificity, 100%). The negative predictive value of ITC was 93.6%, the positive predictive value was 100%, and the accuracy of the method was 93.9%. CONCLUSION: ITC of sentinel nodes is a reliable method. There was no risk of overtreatment due to false-positive results of sentinel node ITC in our study. High accuracy of the method warrants its clinical use. PMID- 18383887 TI - Expression of maspin in papillary Ta/T1 bladder neoplasms. AB - BACKGROUND: The aim of the present study was to evaluate maspin expression in bladder urothelial papillary neoplasms and test the results for correlation with clinicopathological parameters. PATIENTS AND METHODS: A total of 111 urothelial neoplasms from 66 patients were evaluated: pathological examination of primary tumours disclosed 48 pTa and 18pT1. Fourteen additional biopsies, negative for neoplasm, were collected as control biopsies. For each of the 111 neoplastic samples and for the 14 control cases maspin and MIB1 immunoreactivity was evaluated. The immunohistochemical reactions of the 66 primary neoplasms were used for statistical analysis when the disease-free interval, presence and number of relapses, and progression of the disease were tested, whereas all of the 111 tumors were used when the association between the maspin pattern and histological grade and/or pT were evaluated. Thirty-three patients with primary pTa papillary neoplasms (68.7%) and 11 out of eighteen with pT1 (61%), had subsequent relapses of disease. For maspin immunoreactivity the presence/absence of nuclear staining and the pattern of staining were considered. Four patterns of reactivity were recognized and were used for statistical analyses. RESULTS: A statistical association was found between the maspin pattern and pT, histological grade and nuclear staining. CONCLUSION: In papillary urothelial neoplasms, maspin has a pattern of distribution that is associated with the histological grade and pathological stage, and this probably reflects its different activities in the neoplastic process. PMID- 18383888 TI - ER-alpha36, a novel variant of ER-alpha, is expressed in ER-positive and negative human breast carcinomas. AB - BACKGROUND: The status of estrogen receptor-alpha (ER-alpha) expression is one of the most important diagnostic and prognostic factors of breast cancer. ER-alpha is a 66-kDa, ligand-induced transcription factor, characteristically detected in the cell nucleus by immunohistochemistry (IHC) in breast cancer specimens. Recently, we identified and cloned a 36-kDa novel variant of ER-alpha, ER alpha36, which lacks both transactivation domains and functions as a dominant negative effector of transactivation activities of the full-length ER-alpha (ER alpha66) and ER-beta. ER-alpha36 primarily localizes to the cytoplasm and plasma membrane, and responds to both estrogens and antiestrogens by transducing membrane-initiated signaling cascades, stimulating proliferation and possibly contributing to a more aggressive phenotype in breast carcinomas. ER-alpha36 is expressed in established ER-positive and -negative breast cancer cell lines. However, its expression and localization in breast cancer specimens have not been evaluated. As ER-alpha36 may play important roles in breast cancer tumorigenesis, it is of clinical importance to examine the expression pattern of ER-alpha36, in addition to that of ER-alpha66, for more comprehensive molecular profiling of breast carcinomas. PATIENTS AND METHODS: Thirty-one breast cancer patient tissues were evaluated for ER-alpha36 and ER-alpha66 protein expression status by IHC and six additional patient tissue samples were analyzed by Western blot analysis using antibodies specific to ER-alpha66 or ER-alpha36. RESULTS: Our experiments reveal a cytoplasmic and plasma-membrane-associated expression pattern of ER alpha36 in both ER-alpha66-positive and -negative breast cancer samples. Furthermore, ER-alpha36 expression appears to be associated with decreasing nuclear and/or cytoplasmic ER-alpha66 expression, suggesting its potential use as a diagnostic and prognostic marker. CONCLUSION: ER-alpha36 is a novel isoform of ER-alpha, frequently expressed in ER-alpha66-negative cancers, whose detection may provide additional information for better diagnosis and prognosis. PMID- 18383889 TI - Photodynamic therapy of Barrett's esophagus: ablation of Barrett's mucosa and reduction in p53 protein expression after treatment. AB - BACKGROUND: The effectiveness of photodynamic therapy (PDT) for ablation of high grade dysplasia (HGD) in Barrett's esophagus (BE) is typically reported histologically. Following successful PDT, Barrett's mucosa is replaced with neosquamous mucosa. The objective of this study was to compare the expression of p53 protein in neosquamous mucosa as compared to that in HGD samples not treated with PDT. PATIENTS AND METHODS: The patients were divided into two groups. Group I patients (n = 12) had been treated with PDT for HGD and provided 23 biopsy samples of neosquamous mucosa. Group II patients (n = 10) had not received any ablative therapies for BE and provided 14 HGD samples. The immunohistochemical (IHC) staining for p53 protein was performed using mouse anti-human monoclonal antibody DO-1. The degree of p53 protein expression in the cell nuclei was scored using an established IHC scoring system (0 for negative samples and range of 2 to 8 for positive samples). RESULTS: The HGD samples showed diffuse strong p53 staining. The median IHC score for HGD was 7.0. The median IHC score for neosquamous mucosa following PDT was 4.0, with positive scores indicating weak staining in the basal layer of the neosquamous samples. There was significantly lower p53 expression in the neosquamous samples compared to that in the HGD samples (p < 0.001). CONCLUSION: Significantly lower p53 protein expression was detected in neosquamous mucosa of patients who had received PDT for HGD, suggesting a decreased risk for neoplastic progression after treatment. PMID- 18383890 TI - Relationship between oral contraceptive therapy and estrogen receptor status in patients with breast cancer. AB - BACKGROUND: Breast cancer (BC) is the most common cancer in women, and the hormone receptor status is one of the most important prognostic factors in patients with BC. The aim of this study was to establish whether a relationship exists between the hormone receptor rate and the main classic risk factors in patients with BC. PATIENTS AND METHODS: The data regarding a series of 351 consecutive women (median age 57 years, range 26-85 years) who had undergone curative surgery for primary BC was retrospectively reviewed. Eighty-seven (24.8%) patients used oral contraceptives. According to the duration of OC therapy, the patients were dichotomized into two Groups. Group A: less than 22 months (47 patients, 54%) and Group B: 22 months or more (40 patients, 46%). RESULTS: Final pathology showed 15 (4.3%) pT1a, 62 (17.7%) pT1b, 133 (37.9%) pT1c, 125 (35.6%) pT2, and 16 (4.5%) pT3 BC. There were 286 (81.5%) infiltrating ductal, and 24 (6.8%) infiltrating lobular breast carcinomas. The average estrogen receptor (ER) and progesterone receptor (PgR) rate was 59.7 +/- 32.8 and 54.2 +/- 33.9, respectively. There was no relationship (p = NS) between either ER or PgR and the age of the patients, age at menarche and menopause, number of pregnancies, age at first pregnancy, number of spontaneous abortions, months of breastfeeding and the use of estrogen replacement therapy. As expected, ER and PgR rates correlated significantly (R = 0.78, p < 0.01). The ER rates of groups A and B were 51.7 +/- 35.6% and 68.2 +/- 23.6%, respectively (p = 0.014). No other differences (p = NS) between the groups were found. CONCLUSION: The prolonged use of oral contraceptives may increase the ER rate within the tumor tissue, and thus such therapy should be considered an indirect positive prognostic factor in patients with BC. PMID- 18383891 TI - Gemcitabine plus oxaliplatin (GEMOX) in pretreated patients with advanced ovarian cancer: a multicenter phase II study of the Hellenic Oncology Research Group (HORG). AB - The aim of this study was to evaluate the safety and activity of a gemcitabine oxaliplatin combination (GEMOX) regimen in pretreated patients with advanced ovarian cancer. Twenty-seven platinum-refractory/resistant and 14 platinum sensitive patients received gemcitabine 1500 mg/m2 intravenously in days 1+8 and oxaliplatin 130 mg/m2 in day 8 every 21 days. Ten responses (one complete, nine partial) were observed; five in platinum-sensitive and five in platinum-resistant tumors. Grade 3-4 neutropenia and thrombocytopenia were observed in 17 and 10 patients, respectively. There were two treatment-related deaths due to thrombocytopenia and non-neutropenic sepsis. The GEMOX regimen has moderate activity in both platinum-sensitive and resistant ovarian cancer and a toxicity profile that can be significant and therefore requires careful monitoring. PMID- 18383892 TI - Stress proteins HSP27 and HSP70i predict survival in non-small cell lung carcinoma. AB - BACKGROUND: The expression pattern of stress (heat shock) proteins (HSPs) in cancer cells is frequently different from that observed in normal cells; most often some stress-inducible HSPs are constitutively and highly expressed. The objective of this study was to determine the prognostic significance of stress proteins HSP70i and HSP27 in non-small cell lung carcinoma (NSCLC). MATERIALS AND METHODS: An immunohistochemical procedure that enables unambiguous detection of HSP70i protein was used. RESULTS: Strong HSP70i staining showed a survival advantage, although multivariate analysis did not confirm this result. There was an evident correlation between HSP27 overexpression and survival of patients and the results were confirmed by multivariate analysis: 70% of patients with HSP27 negative tumors died within one year after the surgery. CONCLUSION: Our data suggest that HSP27 and HSP70i positivity may represent a favorable prognostic factor in NSCLC. PMID- 18383893 TI - Predictive and prognostic significance of neuron-specific enolase (NSE) in non small cell lung cancer. AB - BACKGROUND: The predictive and prognostic role of neuron-specific enolase (NSE) in non-small cell lung cancer (NSCLC) is still under debate. PATIENTS AND METHODS: To study these aspects, serum NSE was prospectively measured at baseline of first-line chemotherapy treatment and tested for correlation with clinical outcome in 129 advanced NSCLC patients. RESULTS: An objective response was achieved in 27 out of 65 (41.5%) patients with NSE < 8.6 ng/ml and in 38 out of 64 (59.4%) patients with NSE > or = 8.6 ng/ml (p = 0.05). Logistic analysis confirmed the positive association between objective response and NSE values > or = 8.6 ng/ml (odds ratio = 1.69; 95% confidence interval: 1.09-2.63; p = 0.02). Overall median survival was 10.8 months. A statistically significant prognostic effect on survival was found for performance status, stage and response to treatment, but not for baseline NSE value. CONCLUSION: Based on these data, baseline circulating tumor NSE levels appear to have a weak predictive role, but not a prognostic significance in patients with advanced NSCLC submitted to standard chemotherapy. PMID- 18383894 TI - Inguinal sentinel lymph node dissection vs. complete inguinal lymph node dissection in patients with vulvar cancer. AB - BACKGROUND: The aim of this retrospective study was to ascertain the postoperative morbidity in patients with vulvar cancer undergoing sentinel lymph node vs. complete inguinal lymph node dissection. PATIENTS AND METHODS: In total 29 and 46 patients with vulvar cancer, were treated by the technique of inguinal sentinel lymph node dissection or complete inguinal lymph node dissection, respectively. RESULTS: Inguinal sentinel lymph node dissection was associated with a shorter operation time, a reduced rate of inguinal seromas, wound breakdown and wound infection, fewer days of inguinal drainage, and reduced postoperative lymphatic secretion. CONCLUSION: Evidence of reduced peri- and postoperative morbidity with the sentinel lymph node technique for inguinal lymph node dissection in patients with vulvar cancer was demonstrated. PMID- 18383895 TI - Necessity of colposcopy and biopsy prior to large loop excision of the transformation zone (LLETZ). AB - BACKGROUND: The clinical value of colposcopy and biopsy prior to large loop excision of the cervical transformation zone (LLETZ) with respect to negative histology and positive specimen margin rates, was evaluated. PATIENTS AND METHODS: A total of 2,050 consecutive patients who had undergone LLETZ from 1997 to 2005, were included. As diagnostic workup prior to LLETZ, 290 patients underwent repeat PAP test only and 1,760 patients had undergone colposcopy/biopsy. RESULTS: The diagnostic approach using colposcopy/biopsy reduced the rates of negative histology and positive margins from 6.0% to 1.9% (p < 0.001) and from 28.2% to 21.7% (p = 0.002), respectively. The rates of invasive carcinomas were not different between the groups (6.6% vs. 6.5%; p = 0.9). CONCLUSION: Performing colposcopy and biopsy prior to LLETZ reduces unnecessary surgical procedures and decreases positive margin rates. PMID- 18383896 TI - Impact of complementary treatment of breast cancer patients with standardized mistletoe extract during aftercare: a controlled multicenter comparative epidemiological cohort study. AB - OBJECTIVES: To investigate the safety and efficacy of complementary treatment of breast cancer patients with the standardized mistletoe extract (sME) HELIXOR in routine practice during aftercare through a multicenter comparative epidemiological cohort study with 53 randomly selected hospitals/practices representatively distributed in Germany, including oncologists, gynaecologists and general practitioners. PATIENTS AND METHODS: Data from 741 screened patients fulfilling the inclusion/exclusion criteria were checked. Of these, 681 patients were eligible for the final analysis of the study group (with sME n = 167) and the control group (n = 514). Efficacy (development of disease/therapy-induced signs and symptoms; quality of life) and safety (number and severity of adverse events) of complementary treatment in breast cancer patients treated with sME in the aftercare period were determined. RESULTS: Complementary treatment of breast cancer patients with sME during the aftercare period of approximately 5 years after terminating recommended standard therapies resulted in significantly fewer (p < 0.001) complaints of patients (56.3% study group versus 70.0% control group). The reduced number of disease/therapy-related sign/symptoms (e.g. mucositis, fatigue, pain, headache) correlated to a significantly improved quality of life. Adverse drug reactions to the sME treatment were mostly mild and self limiting. CONCLUSION: Complementary treatment with the sME HELIXOR proved to be beneficial for breast cancer patients since it significantly improved quality of life and significantly reduced persistant signs/symptoms of the disease/treatment during the validated aftercare period of approximately five years. PMID- 18383897 TI - Safety and efficacy of zoledronic acid rapid infusion in lung cancer patients with bone metastases: a single institution experience. AB - Zoledronic acid (Zometa, Novartis, Basel, Switzerland) is a new generation of bisphosphonates (BPs) with demonstrated clinical benefit in breast and prostate cancer patients with bone metastases. The safety and efficacy of intravenous zoledronic acid in lung cancer patients was assessed. In 86 patients with newly diagnosed non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC) and bone metastases, 4 mg of zoledronic acid was administered with rapid 15-minute intravenous infusion every 3-4 weeks. A total of 414 infusions were administered over a 24-month period during which a statistically significant decrease in serum calcium levels (p = 0.03) was observed. Serum alkaline phosphatase (ALP) also decreased but not significantly. With regard to clinical efficacy, 55 of our patients stabilized or reduced their need for analgesic treatment. No significant side-effects, including fever, hemodynamic instability and renal dysfunction, were seen. We conclude that the rapid infusion of zoledronic acid is safe and convenient for lung cancer patients even after the 3rd and 6th months follow-up. PMID- 18383898 TI - Clinicopathological features of lipomas with gene fusions involving HMGA2. AB - BACKGROUND: Despite accumulating knowledge of chimeric genes derived from fusion of the HMGA2 gene with multiple partners in lipomas, the different clinicopathological features of lipomas depending on different gene aberrations have not been well documented. The purpose of this study was to examine the clinical significance of the expression of fusion genes in lipomas. PATIENTS AND METHODS: The expressions of three previously reported gene fusion transcripts, including HMGA2/LPP, HMGA2/RDC1 and HMGA2/NFIB, were analyzed in 102 tumors from patients with lipomas. RESULTS: There were 23 cases (22.5%) expressing HMGA2/LPP, 2 cases (1.9%) expressing HMGA2/RDC1 and no cases of HMGA2/NFIB expression (0%). There were no significant intergroup differences in age, gender, body mass index, tumor size or location. The magnetic resonance images and pathological features were also not different in regard to the status of fusion gene expression. CONCLUSION: There were no significant differences of clinicopathological features in patients with lipoma with or without these fusion gene transcripts. PMID- 18383899 TI - Low-dose capecitabine in breast cancer patients with symptomatic bone marrow infiltration: a case study. AB - BACKGROUND: The prognosis of patients with metastatic breast cancer and symptomatic bone marrow involvement is poor. The aim of treatment to these patients is palliation. In this study, we sought to determine the efficacy of therapy with low doses of capecitabine in this subgroup of patients. PATIENTS AND METHODS: Five consecutive breast cancer patients with overt bone marrow involvement were treated by low doses of capecitabine in our department. Four out of five patients also received bisphosphonates to palliate skeletal symptoms. The influence of this therapeutic regimen on tumor response, blood count normalization and overall survival was analysed. RESULTS: All patients except one responded in terms of their haematological profile within two months of the initiation of treatment. Duration of haematological response was 8+ months for all patients. In two of them, tumor response in other sites was evaluated as stable disease, in one as partial remission and in one as progressive disease. Two patients survived more than 22 months without bone marrow failure. CONCLUSION: These initial results are very encouraging for this subset of patients with poor prognosis and limited life expectancy. The administration of capecitabine might be an efficient alternative treatment option. Our results merit further investigation. PMID- 18383900 TI - Prognostic factors in patients with locally advanced (unresectable) or metastatic pancreatic adenocarcinoma: a retrospective analysis. AB - BACKGROUND: Most patients with pancreatic adenocarcinoma are diagnosed with locally advanced (unresectable) or metastatic disease. The aim of this study was to investigate possible prognostic factors of survival in such patients. PATIENTS AND METHODS: Two hundred and fifteen patients were studied retrospectively. Twenty-four potential prognostic variables (demographics, clinical parameters, biochemical markers, treatment modality) were examined. RESULTS: Mean survival was 29.0 weeks. 21.9% survived more than 36 weeks. On multivariate analysis, 10 factors had an independent effect on survival: tumour localisation, metastasis, performance status, jaundice, weight loss, C reactive protein, CEA, CA 19-9, palliative surgery and chemotherapy. Patients managed only with palliative care had a hazard ratio of 8.94 versus those offered a combination of palliative surgery and chemotherapy. CONCLUSION: Many factors could be used as predictors of survival in patients with advanced or metastatic pancreatic cancer. Chemotherapy and palliative surgery are associated with increased survival, and should be offered to all eligible patients. PMID- 18383901 TI - Laryngeal carcinoma recurrence rate and disease-free interval are related to CD105 expression but not to vascular endothelial growth factor 2 (Flk-1/Kdr) expression. AB - BACKGROUND: Tumour angiogenesis is the result of an inbalance between anti- and pro-angiogenic factors. CD105 (endoglin) is a component of the receptor complex of transforming growth factor (TGF-beta1). Vascular endothelial growth factor receptor 2 (VEGFR2 or Flk-1/KDR) belongs to the high-affinity VEGF receptors. The aim of the study was to investigate the expression, cellular localization and role of CD105 and VEGFR2 in laryngeal carcinoma. PATIENTS AND METHODS: Sections of 62 laryngeal carcinomas were stained with CD105 and VEGFR2/Flk-1/KDR antibodies. RESULTS: A significant association between CD105 expression and locoregional recurrence was found (p = 0.009). Interestingly, in N0 patients CD105 expression was significantly associated with locoregional recurrence of the carcinoma (p = 0.03). The log-rank test showed a significant difference in the disease-free interval in patients stratified according to CD105 expression (p = 0.02). Statistical analysis showed no significant associations between vessel endothelial cell or laryngeal carcinoma cell VEGFR2 expressions and recurrence of disease or disease-free intervals. CONCLUSION: CD105 expression but not VEGFR2 expression correlated with carcinoma recurrence after treatment and shorter disease free interval. The CD105 expression may be useful to detect cervical node negative patients with a higher risk of early laryngeal carcinoma recurrence. PMID- 18383903 TI - Amelanotic metastasis of melanoma mimicking ovarian cancer: a case report and review of the literature. AB - Ovarian manifestation of metastatic amelanotic melanoma is exceptionally rare and can lead to the clinical and even histological misdiagnosis of ovarian cancer. We report on a 35-year-old female patient who presented with bilateral adnexal masses, as well as massive ascites. She underwent laparascopy and multiple biopsies were taken. She was histologically diagnosed with malignant ovarian tumour and was referred for radical surgery. Postoperative final histological examination and immunohistochemical staining of the tumour revealed an amelanotic epithelioid melanoma. Despite the variety of this case, clinicians should be aware of this differential diagnosis when treating ovarian cancer. This report discusses the differential diagnosis and clinical management of both metastatic amelanotic malignant melanoma of the ovary and epithelial ovarian cancer. PMID- 18383902 TI - Successful treatment with S-1 plus CPT-11 for lymph node metastasis from colon cancer: report of a case. AB - S-1 is a novel oral anticancer drug, composed of tegafur (FT), gimestat (CDHP) and otastat potassium (Oxo), based on the biochemical modulation of 5 fluorouracil (5-FU). S-1 plus irinotecan (CPT-11) for advanced colorectal cancer as expected showed equally good results as these with CPT-11 plus infusional 5 FU/LV (FOLFIRI regimen). A case of unresectable lymph node metastasis from colon cancer successfully treated with S-1 plus CPT-11 is reported here. A 65-year-old man had metastasis to the lymph nodes in the left supra clavicular region and the superior mesenteric artery. S-1 plus CPT-11 was chosen for the treatment. After 2 courses, since grade 2 toxicity for dysgeusia was observed, S-1 administration was shortened. After 3 courses of the revised regimen, the enlarged lymph nodes disappeared on conventional CT and fluorine-18 fluorodeoxyglucose positron emission tomography with CT (FDG-PET/CT) and the case was assessed as a complete response (CR). Because CR was continued by an additional four courses of treatment, the regimen was changed to a single administration of S-1. Although eighteen months have passed since the induction of CR by S-1 plus CPT-11 therapy, no symptoms or findings of relapse have been observed. PMID- 18383904 TI - Hippocampal sclerosis: histopathology substrate and magnetic resonance imaging. AB - The term hippocampal sclerosis was originally used to describe a shrunken and hardened hippocampus, which histologically displayed neuronal loss and glial proliferation. These alterations are mainly located in the hilus of the dentate gyrus and in the CA1 and CA3 pyramidal cell layers but all hippocampal regions may show neuronal cell loss to varying degrees. A number of morphologic and cytochemical findings are associated with mesial temporal sclerosis, especially within the dentate gyrus. These changes include selective loss of inhibitory interneurons, abnormal sprouting of axons, reorganization of neural transmitter receptors, alterations in second messenger systems, and hyperexcitability of the granule cells. Extrahippocampal pathology is also found at other temporal lobe structures. Frequent extrahippocampal pathology affects the amygdala, first seen with neuronal cell loss and gliosis in the laterobasal complex. Surgical removal of this epileptogenic area can be curative or provide significant reduction in seizure frequency in the majority of individuals. Magnetic resonance imaging (MRI) is highly sensitive in detecting and locating mesial temporal sclerosis when a correct MRI temporal lobe protocol is used. The most important MRI findings, atrophy and abnormal T2 signal, allow us to detect mesial temporal sclerosis in the majority of the cases. Secondary MRI findings help in the diagnosis and lateralization of mesial temporal sclerosis in patients with subtle primary findings and in cases of bilateral hippocampal abnormalities. The development of advanced magnetic resonance (MR) techniques, such as functional MR, diffusion, or transference of magnetization, will lead to greater understanding of this pathology and will improve our diagnostic capacity. PMID- 18383905 TI - Developmental abnormalities of temporal lobe in children. AB - A review of neuroimaging findings in diverse pathologies involving the temporal lobe in children is undertaken. Illustrative images of the developmental disorders that may cause temporal epilepsy in the pediatric context are provided. Brain malformations associated with hippocampal dysgenesis and related conditions such as epilepsy-associated tumors will be discussed with representative examples. Neurocutaneous syndromes that can involve the temporal lobe producing epilepsy or learning defects will be also discussed. Some other miscellaneous disorders occurring preferentially in the pediatric age such as hippocampal damage secondary to hypoxia or radiotherapy will be briefly exposed at the end of the article. PMID- 18383906 TI - MRI of long-term epilepsy-associated tumors. AB - In 20 to 30% of patients with long-term drug-resistant epilepsy neuroepithelial tumors, usually glioneuronal tumors are found. Gangliogliomas and dysembryoplastic neuroepithelial tumors (DNTs) are well characterized, both clinically and on MRI. Both tumor types are located in the cortex or in the cortex and subcortical white matter, gangliogliomas most commonly in the mesial temporal lobe ("around the collateral sulcus"). Both tumor types have typical imaging features, and from both, location and imaging features, they can be usually distinguished from glial tumors. This distinction is important since more than 70% of patients with drug resistant temporal lobe epilepsy caused by gangliogliomas and DNTs get seizure free following extended lesionectomy. PMID- 18383907 TI - Uncommon epiloptogenic lesions affecting the temporal lobe. AB - There are several processes implicated as uncommon causes of temporal lobe epilepsy. Trauma is the leading cause of epilepsy in young adults, intracerebral blood collection being the most consistent risk factor of seizures, especially subdural hematomas and brain contusions. Infarction is the entity most commonly related to epilepsy in the elderly population. Seizures usually present as complex seizures with high recurrence between 6 months and 2 years after stroke. There are some radiological characteristics of the affectation associated with high risk of early and late seizures. Noninfectious limbic encephalitis is a syndrome characterized by seizures, memory loss, and confusion. It includes paraneoplasic and non-paraneoplasic limbic encephalitis, both presenting as hyperintense lesion affecting temporobasal regions more evident with fluid attenuated inversion recovery sequences. Paraneoplasic limbic encephalitis is associated with several types of tumor-induced autoimmunity against the nervous system. The tumors most frequently implicated are the lungs, testis, and breast, including Hodgkin's lymphoma, teratoma, and thymoma in young patients. Once a tumor is excluded, non-paraneoplasic limbic encephalitis has to be considered by investigating the presence of antibodies against voltage-gated potassium channels. It is associated with hyponatremia and responds to regimens of steroids, plasma exchange, and intravenous immunoglobulins. Finally, herpetic limbic encephalitis is also associated with seizures, accompanied by fever and neurologic symptoms. It presents characteristic findings and distribution on magnetic resonance imaging, which shows abnormalities in more than 90% of patients with proven Herpes simplex virus type 1. PMID- 18383909 TI - Taking care of patients also means taking care of business. PMID- 18383910 TI - Hospital's plan assures correct admission status. PMID- 18383908 TI - Predictors, procedures, and perspective for temporal lobe epilepsy surgery. AB - Surgery for refractory temporal lobe epilepsy is established as a promising treatment option. Although practiced for decades now, there is an ongoing discussion on who is a candidate, when we should operate, how the preoperative evaluation and surgery should be performed, and what perspective will result for the individual after surgery. In light of significant improvements with respect to safety and outcome, generally accepted features will be extracted, and some more ambivalent aspects will be discussed. Major future developments in this field will be based on multimodal imaging and correlation with electro-clinical and neuropsychological findings. PMID- 18383911 TI - Don't neglect contributing factors in assessment. PMID- 18383912 TI - Discharge disposition initiative increases reimbursement. PMID- 18383913 TI - Web site links to valued post-discharge resources. PMID- 18383914 TI - Department takes control of managed care denials. PMID- 18383916 TI - Pregnant patient at risk for unnecessary appendectomy. PMID- 18383915 TI - Decrease readmissions with 'whole-family' approach. PMID- 18383917 TI - [Signal pathways of cell proliferation and death as targets of potential chemotherapeutics]. AB - The purpose of this paper is to review current information concerning signal transduction pathways of cell proliferation and cell death applicable in the research of antitumor compounds with a specific effect. Actually, cancer counts among the world gravest diseases. Research of the mechanisms of action of chemotherapeutics helps us to find compounds with high cytotoxic activity to tumor cells and low or no cytotoxicity to normal cells. Many present studies deal with the ability of drugs to hit the proliferation signal pathways or cell death pathways specifically. Various proliferation signal pathways have been identified, e.g. pathways of mitogen-activated proteinkinases. In original studies, cell death was considered to perform in necrotic and apoptotic forms, whereas in contrast to necrosis, apoptosis represented the programmed process. However, other forms of programmed cell death were discovered, the programmed necrosis and autophagic cell death. Similarly, beside the intrinsic, mitochondrial-mediated, and extrinsic, receptor-mediated pathways, new mechanisms of induction of apoptosis were discovered: the endoplasmic reticulum stress pathway in which calcium plays an important role, the lysosomal pathway and the ceramide-induced pathway. Current information concerning transduction of antiproliferative and death stimuli in cells allows to explain the mechanisms of action of known drugs and also brings novel therapeutical targets which can serve in treatment of such diseases as cancer. PMID- 18383918 TI - [Hemostatic effects of oxidized cellulose]. AB - Oxidized cellulose ranks among nontoxic and biocompatible biopolymers. Oxidized regenerated cellulose (ORC) is manufactured from regenerated cellulose derived from wood pulp containing about 50% of cellulose. To obtain purified cellulose, it is necessary to decompose it in a chemical way and subsequently put it together to make "regenerated" cellulose. Thanks to its good hemostatic effects, high biosolubility and biodegradability, antioxidant and wound-healing properties, oxidized cellulose represents a suitable means for the therapy of bleeding conditions in various fields of medicine. In addition, the confirmed bactericidal effects of oxidized cellulose towards a wide spectrum of aerobic and anaerobic pathogens increase the therapeutic potential of this agent for use in clinical practice. At present there is a renewed interest in its wider use in clinical practice and in an improvement of the knowledge of its mechanisms of effects, which are tested in vitro, on animal models as well as in clinical studies. The present paper attempts to summarize the hitherto knowledge of hemostatic properties of oxidized cellulose and also to characterize other possible biological effects. PMID- 18383919 TI - Effects of auxins on growth and scopoletin accumulation in cell suspension cultures of Angelica archangelica L. AB - Scopoletin is a coumarin possessing many interesting biological effects, e.g., spasmolytic, anti-inflammatory, antimutagenic, antioxidant, antifungal, apoptosis inducing, antiproliferative, acetylcholinesterase-inhibitory, and hypouricemic activities. Plant tissue cultures represent a promising alternative source of valuable plant-derived substances. A number of physical and chemical factors influence the cell growth and secondary metabolite biosynthesis in plant tissue cultures. The mechanism of their action is not completely understood. Besides other factors, plant growth regulators and light conditions play an important role. Effects of four auxins (2,4-dichlorophenoxyacetic acid, 2,4-D, alpha naphthaleneacetic acid, NAA, beta-indoleacetic acid, IAA or beta-indolebutyric acid, IBA) at four concentrations (0.2, 2, 10 or 20 mg/l) on the culture growth and accumulation of scopoletin in the medium were tested in Angelica archangelica cell suspension cultures cultured under continuous light or in the dark. The highest culture growth was achieved with 2 mg/l 2,4-D, and 10 mg/l IAA. The best scopoletin levels were obtained with 0.2 mg/l 2,4-D, 2 mg/l 2,4-D, 10 mg/l NAA, and 20 mg/l IAA. The effects of light conditions were less marked than those of auxins and their concentrations in influencing both the cell growth and scopoletin accumulation in Angelica archangelica cell suspension cultures. The changes brought about by auxins were modified by light conditions. PMID- 18383920 TI - A study of the properties of tablets made of directly compressible maltose. AB - The paper deals with the study of the strength and disintegration time of tablets made of directly compressible maltose Advantose 100. It studies the differences of the effects of two types of lubricants, magnesium stearate and sodium stearylfumarate, on the above-mentioned properties, and it also tests the mixtures of the substance with microcrystalline cellulose Vivapur 102 in a ratio of 1:1 and with ascorbic and acetylsalicylic acids. The compacts are obtained by using three compression forces, excepting mixtures with active ingredients, where one compression force is used. In the compression forces of 6 and 8 kN, no statistically significant difference was found in the intervention of the lubricants into the strength of the compacts made of Advantose 100, only in the compression force of 10 kN Pruv decreased the strength more than stearate. The mixture of Advantose 100 and Vivapur 102 yielded the strongest tablets, an addition of Pruv to it decreased the strength of compacts more than stearate. The periods of disintegration time of Advantose compacts as well as those of the mixture of dry binders were longer with an addition of Pruv. The compacts with acetylsalicylic acid possessed higher strength and a longer period of disintegration than those with ascorbic acid. There was no statistically significant difference within the type of the lubricant employed, both in the case of Advantose 100 and its mixture with Vivapur 102, between the values of strength of the compacts with acetylsalicylic acid. PMID- 18383921 TI - [Substances modifying the activity of caspases]. AB - Caspases are enzymes which play a key role in programmed cell death--apoptosis. Dysregulation of this process results in a series of disorders in which apoptosis is involved in pathogenesis. Diseases could be divided into two groups--diseases with pathological inhibition of apoptosis (cancer, some autoimmune disorders) and diseases with pathological induction of apoptosis (neurodegenerative disorders, AIDS). The paper lists the most significant activators and inhibitors of caspases as they control apoptosis and hence they are widely studied nowadays. The effects of stobadine, theophylline and adenine derivatives on the activity of caspase 1 was investigated with the use of spectrophotometry. The compounds under study showed an inhibitory effect on the enzyme tested; in one case the inhibitory effect exceeded 80%. PMID- 18383922 TI - [Antioxidant activity of tinctures prepared from hawthorn fruits and motherwort herb]. AB - The paper aimed to determine and compare the antioxidant activity of two tinctures and selected flavonoids present in the tinctures. The tinctures were prepared from hawthorn fruits and motherwort herb. The group of selected flavonoids included rutin, quercetin, hyperosid, epicatechine, and procyanidin B2. Two methods were employed to determine antioxidant activities: the inhibition of peroxynitrite-induced nitration of tyrosine was measured, and the ability to quench radicals was examined by of the stable radical 2,2-diphenyl-1 picrylhydrazyl (DPPH). Both tinctures under study were able to quench radicals and reactive forms of oxygen and nitrogen. The motherwort herb tincture was more effective, but no direct effect of phenolic compounds on the extent of antioxidant activity was demonstrated. The total content of polyphenols in the motherwort herb tincture was higher by 163 %, nevertheless antiperoxynitrite activity was higher just by 19 % (p < 0.05), and the ability to reduce DPPH differed in both tinctures in a minimal manner. Of the individual flavonoids, the substances present in the tinctures in higher concentrations contributed most to antioxidant activity. In the hawthorn fruit tincture it was epicatechine and hyperosid, whereas in the motherwort herb tincture it was rutin. PMID- 18383923 TI - A report from the department of staph affairs. PMID- 18383924 TI - Medical causes of back pain. PMID- 18383925 TI - What is adequate hypertension control? PMID- 18383926 TI - What are the caveats to using sodium phosphate agents for bowel preparation? PMID- 18383928 TI - What can patients expect from cataract surgery? AB - Cataract surgery has evolved into an outpatient procedure that requires minimal anesthesia and significantly improves visual function for about 90% of patients. With the help of their primary care physician and ophthalmologist, patients can decide when cataract surgery is appropriate for them. In addition, patients may have a choice about the type of synthetic lens implant that fits their visual needs. PMID- 18383927 TI - Staphylococcus aureus: the new adventures of a legendary pathogen. AB - Nosocomial infections with strains of methicillin-resistant Staphylococcus aureus (MRSA) began to emerge in the 1960s, are increasing in frequency, and tend to have worse outcomes than infections due to methicillin-susceptible S aureus. Community-associated MRSA infections emerged in the 1990s. Community-associated MRSA strains have up to now been epidemiologically and bacteriologically distinct from hospital-associated MRSA strains, but in a new twist, MRSA strains that have sofar been only community-associated are invading the hospital. Another worrisome trend is increasing resistance to vancomycin (Vancocin). PMID- 18383929 TI - Should patients on long-term warfarin take aspirin for heart disease? PMID- 18383930 TI - A young woman with fatigue. Celiac disease. PMID- 18383931 TI - How to evaluate 'dipstick hematuria': what to do before you refer. AB - Although major health organizations do not support screening for hematuria by dipstick testing, millions of patients without symptoms are tested yearly. Since urinary dipstick tests for hematuria have a high false-positive rate, patients with positive dipstick results require microscopic urinalysis before the diagnosis of hematuria can be made. Primary care physicians can help protect patients from the anxiety, costs, and risks of an unnecessary urologic workup by adhering to the principles of early hematuria management. PMID- 18383932 TI - Preventing a first episode of esophageal variceal hemorrhage. AB - In patients with esophageal varices, hemorrhage is common and often lethal, so we need to take a proactive approach to preventing a first episode of bleeding. All patients with cirrhosis should undergo endoscopy to look for varices. Depending on the size and appearance of the varices and the patient's Child-Pugh grade, prophylactic treatment may be indicated. PMID- 18383933 TI - ["The union has the power"!]. PMID- 18383934 TI - [Towards a single society for specialists in family medicine and specialists in general internal medicine in Switzerland?]. AB - The debate on the legimity of having two distinct medical societies and specialist degrees in Switzerland, one for family medicine and the other for internal medicine, is currently reactivated due to the rapid evolution of medical demographics and of our health system. In this paper, the academic representatives of those disciplines from the French-speaking part of Switzerland sum up the arguments for and against creating a single medical society for generalists and internists, in the perspective of education and research. They conclude that the advantages vastly outweigh the inconveniences and support this project. Moreover, they propose a training track individualized according to the trainee's future practice environment (hospital-based, urban or suburban outpatient practice). PMID- 18383935 TI - [Highlights 2007 in hospital-based internal medicine: the point of view from the chief residents]. AB - In 2007, several international studies brought useful information for the daily work of internists in hospital settings. This summary is of course subjective but reflects the interests and questions of the chief residents of the Department of internal medicine who wrote this article like an original trip in medical literature. This trip will allow you to review some aspects of important fields such as heart failure, diabetes, endocarditis, COPD, and quality of care. Besides the growing diversity of the fields covered by internal medicine, these various topics underline also the uncertainty internists have to face in a practice directed towards evidence. PMID- 18383936 TI - [Update in hospital internal medicine (2007): a selection]. AB - This paper summarizes several important studies published during the previous year that have an impact on the practice of inpatient internal medicine, because they either modify or reinforce current practices. The selected domains include the treatment of thrombo-embolic disease, the role of implantable defibrillators in left cardiac failure, the management of cerebro-vascular disease, of community acquired pneumonia, and of type 2 diabetes, as well as the prevention of spontaneous bacterial peritonitis in cirrhosis and of osteoporosis fractures. Some data stimulating our reflection on the integration of medical education in health care centres and on the validity of some scientific publications are also presented. PMID- 18383937 TI - [Innovations in ambulatory care: screening, treatment and iatrogenicity]. AB - This year we have selected six studies useful for the day to day practice. A mammography in women 40 to 49 years of age should be evaluated taking into account the patient's profile and the possible risks and benefits of this exam. In patients over 65 years of age, a systematic atrial fibrillation screening, with pulse rate measuring then ECG (if irregular beat) should be realised on a regular basis. The risks for complications following colonoscopies do exist, especially after biopsies/polypectomies and this risk should be discussed. Long term proton pump inhibitor treatment is a risk factor for hip fracture. It is important to treat high blood pressure problems in the elderly, but the orthostatic risks should be adressed. A corticoid treatment started quickly for Bell's palsy is efficient. PMID- 18383938 TI - [Relevant publications in ambulatory general internal medicine in 2007]. AB - Screening procedures for genital Chlamydia infection, cancer risks linked to oral contraceptives, indications and efficacy of HPV vaccination, and diagnostic tools for celiac disease in adults; these are just a few of the general practice themes that were reviewed and analysed in 2007 by residents and chief residents at the Community medicine and primary care Service of the Geneva University Hospitals. These commented summaries, intended for all our colleagues, constitute Geneva's contribution to the literature data base initiated in 2005 by chief residents in Lausanne. PMID- 18383939 TI - [Review on abdominal aortic aneurysm screening]. AB - Health organizations recently recommended one-time screening for abdominal aortic aneurysm (AAA) by ultrasonography in men aged 65 to 75 who have ever smoked. This recommendation emerged from the results of a systematic review showing that AAA screening and surgical repair of large AAA leads to a decreased AAA-specific mortality in this population. Moreover, AAA screening is cost-effective. However, concerns persist about the effectiveness of AAA screening when applied in the daily practice setting, and the best care of small AAA for which research clinical trials have shown that early, elective surgery does not save lives. PMID- 18383940 TI - [Deconstructing emergency psychiatry: a program and a Neuchatel practice]. PMID- 18383941 TI - [The trafficking of human beings in Switzerland: what role for primary care physician?]. PMID- 18383942 TI - [Food and milk clones declared]. PMID- 18383943 TI - [What is a possible human case of avian flu?]. PMID- 18383944 TI - [Occupational stress and coronary pathology]. PMID- 18383945 TI - [Little Switzerland]. PMID- 18383946 TI - Did nurse give Digoxin O/D which caused death? PMID- 18383947 TI - Don't let Dr. 'pass the buck' for disposal of sharps. PMID- 18383948 TI - NJ: UI benefits denied-payments made in error: nurse ordered to repay all benefits received. PMID- 18383949 TI - Did infiltrated injection site call for D/C of taxol infusion? PMID- 18383950 TI - Globalisation of healthcare--a reality or distant dream. PMID- 18383951 TI - Assessmrent of iron overload in thalassaemic patients by magnetic resonance imaging: a pilot study. AB - The patients of thalassaemia major need repeated blood transfusion which leads to excess iron deposition in various organs like liver, heart, pituitary etc. This iron accumulation causes various complications and ultimately organs' failure. There is no non-invasive, standard and reliable method to know the status of iron overload in various organs of the body. This paper attempts to use magnetic resonance imaging to know the liver iron overload in 8 thalassaemic patients as a pilot study. Eight children suffering with thalassaemia and 3 controls who were the normal siblings of the patient group underwent magnetic resonance imaging of the abdomen using spin-echo T, weighted sequence. Blood serum ferritin levels in the patients' group were also determined on the same day of magnetic resonance imaging examination. It was observed that the ratio of magnetic resonance imaging signal intensity (in spin-echo T1 weighted image) in paraspinous muscle to liver was significantly different in normal control (0.65) compared to that in thalassaemia patients (2.1 to 11.4 depending upon extent of iron deposition). The magnetic resonance signal intensity ratio correlated with the blood serum ferritin level of patients (p = 0.01) which is generally taken as indirect measure of body iron burden. Spin-echo sequence is the simplest imaging sequence and it increases the chance of its routine use. The study concludes that magnetic resonance imaging has good potential to quantify the liver iron deposition non invasively and may denote the efficacy of iron-chelation therapy which is used to reduce the body iron burden in these patients. PMID- 18383952 TI - A hospital-based study on pulmonary function tests and exercise tolerance in patients of chronic obstructive pulmonary disease and other diseases. AB - A total of 105 male and 60 female patients were screened in the respiratory medicine outpatients' department, Institute of Postgraduate Medical Education & Research, Kolkata between December, 2002 and January, 2005. Chronic obstructive pulmonary disease patients were diagnosed on the basis of history and clinical examination while patients with body mass index > or =25 and otherwise disease free were grouped as overweights. Patients suffering from other diseases like systemic hypertension, etc, were referred from other departments after proper evaluation. The patients and controls (n=10) each for male and female groups were subjected to spirometry using computerised electronic spirometer while exercise tolerance was evaluated by modified Harvard step test. Thirty-nine male and 21 female patients were diagnosed and grouped in chronic obstructive pulmonary disease group. Both purely obstructive [forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC)% pred<70 and reduced FEV1% pred] and mixed (both obstructive and restrictive pattern ie, FEV1/FVC% pred normal or supernormal, FVC% pred <80 indicating restrictive pattern and forced expiratory flow between 25% and 75% of the vital capacity (FEF(25-75)) pred or peak expiratory flow rate (PEFR)% pred <70 indicating early small airway obstruction pattern were seen in both sexes. Although the exercise tolerance values were non significant in both sexes in chronic obstructive pulmonary disease obstructive pattern group, in mixed pattern group it was seen significant reduction compared to control. Hypertensives (21 males and 7 females) showed obstructive spirometric pattern. Exercise tolerance values were significantly reduced compared to controls. Male overweights (n=13) showed restrictive pattern while female overweights (n=8) showed obstructive pattern in spirometry. Exercise tolerance values were non-significant compared to control in both the groups. In ischaemic heart disease patients (n=6) FEV1%pred showed significant reduction in spirometry. In patients suffering from type 2 diabetes mellitus (n=4), post tuberculous group (n=7), hypothyroid (n=6), collagen vascular disease group (n=6) showed restrictive spirometric pattern and the above groups including IHD patients showed significant reduction in exercise tolerance values. Some authors have stated that mixed ventilatory defect is characterised by low FEV1/FVC% pred in spirometry and low lung volumes where the lung volumes have to be ascertained by other methods but in the present investigation it was observed that mixed ventilatory defect can be estimated by spirometry; PEFR and/or FEF(25-75%). pred <70% whereas FEV1/FVC% pred is normal or supernormal. This finding is completely new one to predict mixed ventilatory defect. PMID- 18383953 TI - Current pattern of enteric fever: a prospective clinical and microbiological study. AB - A prospective clinical and microbiological study was conducted in 145 blood culture positive cases of enteric fever below the age of 18 years over a period of eleven months (June 2004 to April 2005). It aimed to study the clinical profile, the relative magnitude of enteric fever in children, especially in those below the age of two years and to determine the current antibiotic sensitivity pattern of Salmonella typhi and S paratyphi. Enteric fever is a significant problem in the preschool years. Sixty-five per cent of cases were in the age group of 2 to 9 years, 27% in 0-5 years and 13% in age group 0-2 years. Ninety two per cent of the cases were caused by S typhi. Paratyphoid fever is less common (8%), when occurs is caused by S paratyphi A. In-vitro sensitivity, using the Bauer-Kirby agar disc diffusion method, to ceftriaxone was 99%, cefixime-99%, cefotaxime-99%, cefpodoxime-72%, cefoperazone-93%, among quinolones, ciprofloxacin-95%, ofloxacin-83%, norfloxacin-79%. Sensitivity to the originally used antibiotics is reappearing: Ampicillin-87%, amoxicillin-89%, trimethoprim sulfamethoxazole-76%, chloramphenicol-86%. Among other drugs, sensitivity to imepenam-100%, azithromycin-49%, aztreonam-65%, amikacin-98%. Nalidixic acid resistance is very high ie, 88%. PMID- 18383954 TI - A study on nephropathy in type 2 diabetes mellitus: histology and its correlation with clinical and biochemical parameters. AB - To document clinical, serum and urinary parameters in patients with features of diabetic nephropathy in type 2 diabetes mellitus and to correlate light microscopical findings of the renal biopsy specimen with the clinical, serum and urinary parameters, a study was conducted among 30 patients of type 2 diabetes mellitus with features of nephropathy attended Medical College, Kolkata with special emphasis given on neurological and fundoscopic examination. The patients were investigated with fasting and postprandial venous plasma glucose, glycosylated haemoglobin (HbA1c), serum urea, creatinine, sodium, potassium, 24 hour urinary protein/micro-albumin and lipid profile. Percutaneous renal biopsy was performed after taking informed consent from the patients and sent for histopathological examination. Obtained data were tabulated and analysed. Among 30 patients (male-16; female-14; proteinuric-23, micro-albuminuric-6) diabetic nephropathy was detected in 28 patients (diffuse-15, nodular-9, 4 had focal segmental glomerulosclerosis) and 2 had normal renal biopsy. Retinopathy was detected in 16 patients; 12 had autonomic neuropathy and 3 had peripheral neuropathy. PMID- 18383955 TI - Unilateral lower limb oedema in pregnancy. AB - A 21 years old lady was admitted with severe pain and swelling of left lower limb in a pregnancy of 34 weeks of period of gestation. On examination there was pitting oedema of the whole left lower limb with extreme tenderness. Doppler USG of left femoral vein showed a thrombus in the process of recanalisation and no significant flow was demonstrated inside the lumen and USG for foetoplacental profile showed a single live foetus of gestational age 34 weeks. The case was diagnosed as deep venous thrombosis in pregnancy. She was treated with heparin injection 5000 unit subcutaneously twice daily throughout pregnancy along with antibiotics. In spite of proper management she developed extensive ulceration in left lower limb. She delivered normally a healty male child after induction with oxytocin dirp. The postpartum period was uneventful. Throughout this period heparin was continued and she was referred to surgery department for skin grafting. PMID- 18383956 TI - [Metabolic syndrome and depression--clinical relations]. AB - The occurrence of both obesity and type 2 diabetes mellitus is rapidly increasing; according to WHO data, this can be considered as a worldwide epidemic. The obesity is one of the components of metabolic syndrome, the cluster of several risk factors of atherosclerosis such as dyslipidemia, hypertension, impaired glucose homeostasis, pro-thrombotic state and subclinical inflammation. The importance of the metabolic syndrome is confirmed by findings of the several times increased risk of both the type 2 diabetes mellitus and cardiovascular disease. Similarly, as in the case of obesity and diabetes, the incidence and prevalence of depressive disorder are still increasing and depressive disorder belongs to the most important causes of disability. The interrelations between depressive disorder and diabetes are known for a long time. Diabetics very often suffer from depression and vice versa, the depressive disorder is a significant risk factor of type 2 diabetes mellitus development and worsens the survival of diabetics. Those relationships have been recently intensively studied. Our paper reviews genetic, nutritional, metabolic and hormonal factors, contributing to the above mentioned syndrome. PMID- 18383957 TI - [Metabolic effects of the endocannabinoid system]. AB - The endocannabinoid system (ECS) is an endogenous signalling system that plays an important role in the regulation of energy homeostasis and lipid and glucose metabolism-all of which can influence cardiometabolic risk. Endocannabinoids are involved in the appetite and body weight regulation. Central or peripheral stimulation of ECS favours metabolic processes that lead to weight gain, lipogenesis, insulin resistance, dyslipidemia, and impaired glucose homeostasis. ECS is tonically overactive in obese individuals. ECS appears to be a promising novel mechanistic pathway that modulates important aspects of the cardiovascular and metabolic function. PMID- 18383958 TI - [Semen quality of Pilsner University students]. AB - BACKGROUND: Decreasing quality of the semen represents a big problem for human population. Parameters of spermiogramms are used for obtaining of information about the human semen quality. The aim of our study was to evaluate parameters of semen and its immunological characteristics in relation to data from personal history of our students. METHODS AND RESULTS: Basic analysis of semen samples of 110 students was performed according to World Health Organization criteria. Direct mixed-antiimmunoglobulin-reaction-tests for detection of sperm agglutinating antibodies, intraacrosomal proteins by specific monoclonal antibodies and immunoflurescence examination were used. Data from anamnesis were obtained by a questionnaire. Normozoospermia was found in 72% cases, oligozoospermia in 14%, asthenozoospermia in 5%, teratozoospermia in 4%, oligoasthenozoospermia in 5% of men. We have not confirmed negative influence of smoking, drug abuse, wearing tight clothes, hot baths, and using sauna on sperm quality. There was significantly higher percentage of students consuming presevered and spicy food more than twice a week in the group with pathological spermiogramm in comparison with the normozoospermic group. Acrosomal proteins were found in 75% men with normozoospermia and in 43% of pathological spermiogramms. Findings of sperm-agglutinating antibodies were not significant. CONCLUSIONS: Semen quantity as well as quality is affected by many intrinsic and extrinsic factors. Our study shows that fertility of our students is better than we expected. PMID- 18383959 TI - [Brain natriuretic peptide might predict the abnormal geometry of heart in children with chronic kidney disease]. AB - BACKGROUND: Cardiovascular diseases are the leading course of morbidity/mortality in children with chronic kidney disease. Recently it has been shown that a brain natriuretic peptide is a sensitive cardiac marker for classification of the cardiovascular risk in adults. Whether it has the same diagnostic value in children with chronic kidney diseases has to be established. The purpose of the study was to evaluate whether the brain natriuretic peptide can predict cardiac dysfunction in children with chronic kidney disease. METHODS AND RESULTS: Relation between serum level of the brain natriuretic peptide, echocardiography and cardiovascular risk factors (hypertension, anaemia, lipids, C-reactive protein, secondary hyperparathyreoidism) has been studied in 46 children (10 patients at predialysis, 14 patients on dialysis, 11 children with kidney transplant and 11 healthy controls). Brain natriuretic peptide was significantly higher in dialysed patients (2.09 +/- 0.78) in comparison with healthy children (1.43 +/- 0.34, p = 0.012) and with both groups of patients at pre-dialysis stage (1.52 +/- 0.42, p = 0.039) and after kidney transplant (1.71 +/- 0.46, p = 0.19). Abnormal heart geometry was found in 19 patients (54.28%). Compared to controls, brain natriuretic peptide was higher in children with eccentric but not in those with concentric hypertrophy (2.178 +/- 0.956 vs. 1.496 +/- 0.395, p = 0.05, resp. 1.982 +/- 0.618 vs. 1.496 +/- 0.395, p = 0.04). Significant correlation was found between levels of brain natriuretic peptide and ventricular hypertrophy (p = 0.001), with the level of parathyroid hormone (p = 0.03) and with the degree of anaemia (p = 0.027). CONCLUSIONS: Brain natriuretic protein can predict an abnormal geometry of heart in children with chronic kidney disease. Our preliminary results suggest that it is a suitable marker of cardiovascular classification in paediatrics. PMID- 18383960 TI - [Tracheal intubation without muscle relaxant--the impact of different sufentanil doses on the quality of intubating conditions: a prospective study]. AB - BACKGROUND: Following opioid and intravenous anaesthetic, tracheal intubation may be accomplished without any muscle relaxant. In this study, we evaluated the dose requirements for sufentanil with propofol without muscle relaxant administration to obtain clinically acceptable intubation conditions and cardiovascular responses. METHODS AND RESULTS: After midazolam (0.05 mg x kg(-1)), 90 patients were randomized into three equal groups, each receiving sufentanil 0.2 (SUF-0.2), 0.3 (SUF-0.3) or 0.4 (SUF-0.4) microg x kg(-1), respectively. Following induction of anaesthesia with propofol (2 mg x kg(-1)), the quality of tracheal intubation was assessed as excellent or good (clinically acceptable) and poor. Excellent intubation conditions were observed in 28%, 41% and 54% of patients in groups SUF 0.2, SUF-0.3 and SUF-0.4, respectively, while poor conditions were present in 31% (SUF-0.2; p = 0.041 vs. SUF-0.3, p = 0.006 vs SUF-0.4), 7% (SUF-0.3) and 3% (SUF 0.4). Patients with induction-related hypotension were older than those without (60 +/- 15 vs. 52 +/- 17 yr, p = 0.013). CONCLUSIONS: Sufentanil (0.3-0.4 microg x kg(-1)) in combination with propofol (2 mg.kg(-1)) provided clinically acceptable intubating conditions in 93-97% patients. PMID- 18383961 TI - [Complications with the pocket of cardiostimulator]. AB - BACKGROUND: The number of patients with pacemakers is icreasing. The first reason is longer survival, the second reason is widening indications to cardiostimulation. Along with increasing numbers of performances, we have to recken larger number of complications and these complications solve so, that we avoid more serious damage of a patient. METHODS AND RESULTS: In our article we focused an possible complications in the area of pacemaker pockets, monitored their appearence, watched the type of complication, the danger of the creation infection endocarditis and we present our own experiences with solving such complications. From the total numer of 3978 surgical interventions was 141 patients indicated for the revision of the pocket. CONCLUSIONS: Repeated controls of the pocket and early reported of the patients to the specialised clinic is necessary for the safety of tke patients with pacemakers. PMID- 18383962 TI - [Experience with early enteral nutrition application in critically ill patients in medical intensive care unit]. AB - BACKGROUND: Early enteral nutrition is recommended in patients with critical illness. Generally implementing of nutritional support algorithm is advised. The aim of study was evaluation of early enteral nutrition application in critically ill patients in medical intensive care unit. METHODS AND RESULTS: Early enteral nutrition was given according to written protocol in medical intensive care unit. During the first 96 hours hypocaloric nutrition 20-25 calories/kg was applied, followed by increase to 25-30 calories/kg at the end of the first week of admission. Apart from the patients who reached 25-30 calories/kg we recorded the number of patients who tolerated hypocaloric enteral nutrition and evaluated the number of patients with complications due to enteral nutrition. Early enteral nutrition was given to 44 out of 99 patients admitted to intensive care unit with life threatening diasese and indication for nutrition support. Out of 44 critically ill patients (35 with sepsis, 9 with another medical emergency) 22 died during admisssion in intensive care unit (50%). Hypocaloric enteral nutrition during the first 96 hours was given to 36 patients (82%). In 8 patients enteral nutrition had to be stopped and substituted for parenteral one due to complications. Three patients suffered from abdominal distension, 2 from profused diarrhea, 1 from combination of diarrhea and abdominal distension and 2 from aspiration. Twenty seven patients tolerated the application of enteral nutrition via nasogastric tube. In 10 patients nasogastric tube had to be replaced for nasojejunal one for high gastric aspirate volume. The caloric intake of 25-30 calories/kg was reached by the end of the first week of admission in 26 patients (60%). CONCLUSIONS: Early enteral nutrition applied according to protocol was given succesfully to the substantial number of the critical patients. In 18% of the patients enteral nutrition had to be replaced for parenteral one due to complications. The caloric intake 25-30 calories/kg was reached in 60% of patients. PMID- 18383963 TI - [Is the extent of present examination of blood donors sufficient? Malignancies in active blood donors]. AB - Casuistry brings the evidence that the present extent of examination before each blood donation is not sufficient for the identification of malignancies. Donation of blood can coincide with the development of tumour into a far gone stage or into generalization. Sensitivity of the pre-extraction examination for the presence of malignancy is low. PMID- 18383964 TI - [The occurrence of malignancies in former blood donors]. AB - Occurrence of malignancies in the group of 267 former blood donors who donated blood 20 times and more (multiple donors) were compared with those who donated blood 10 times or less. Cohorts were comparable in age, period from the first donation and ratio of blood groups in A, B, 0 system. In the group of multiple donors, 31 (11.6%) persons were listed in the oncological register for 36 malignancies. Among the less frequent donors only 14 persons (5.3%) were listed with 15 malignancies. The difference in malignancy incidence between both groups is highly significant. The difference is significant for the cohorts of men, being not significant for women; probably because of small number of cases. Our results indicate the possibility of carcinogenic effect of the blood donation. PMID- 18383965 TI - [Second Department of Internal Medicine, Masaryk University. Faculty of Medicine at Sant Anna in Brno is celebrating the 60th anniversary of the foundation]. PMID- 18383966 TI - [Lipid emulsion for the treatment of severe local anesthetic toxicity in adults- probably useful, but evidence is lacking]. PMID- 18383967 TI - [Anesthetic induction with nitrous-oxide-free sevoflurane in pediatric patients]. AB - OBJECTIVE: To evaluate the pediatric use of inhaled nitrous oxide (N2O)-free induction with sevoflurane for the purpose of protecting staff from exposure to workplace air pollution. PATIENTS AND METHODS: Prospective, randomized trial in ASA class 1-2 children in whom a tidal breathing technique was used for anesthetic induction in a variety of surgical procedures. Patients were allocated to 2 groups. The sevo-N2O group inhaled 8% sevoflurane in a 60/40% mixture of oxygen and N2O. The sevo-air group received 8% sevoflurane in a mixture of oxygen and air (inspired oxygen fraction, 40%). We recorded mean arterial pressure (MAP), heart rate, oxygen saturation by pulse oximetry (SpO2), limb response to venous puncture, alveolar concentration of sevoflurane, and incidence of adverse events. RESULTS: Twenty-two patients were assigned to each group. The vein was catheterized in all patients without a pain reflex in the limb, and there were no statistically significant differences in MAP, heart rate, SpO2, or incidence of adverse events. Mean (SD) alveolar concentration of sevoflurane, however, differed between the 2 groups: 53% (0.51%) in the sevo-N2O group and 4.91% (0.41%) in the sevo-air group (P = .028). CONCLUSIONS: N2O-free anesthetic induction by tidal breathing of 8% sevoflurane provides similar anesthetic conditions (efficacy, safety, and rapid onset) without a higher incidence of adverse events. The use of N2O can therefore be avoided. PMID- 18383968 TI - [Effectiveness of tranexamic acid in routine performance of total knee replacement surgery]. AB - OBJECTIVES: To evaluate the effectiveness of treatment with tranexamic acid, compared to absence of antifibrinolytic treatment, in reducing transfusion rates and the number of units of packed red blood cells required in patients undergoing total knee replacement surgery. MATERIAL AND METHODS: We reviewed the medical records of all patients who underwent total knee replacement surgery in a general hospital in 2006. Information was recorded on treatment with tranexamic acid, use of other antifibrinolytic drugs, hemoglobin and hematocrit levels before surgery and 3 days after surgery, patients requiring transfusions, units of packed red blood cells administered, and whether or not drains were clamped within 4 hours. Complications attributable to tranexamic acid (thromboembolic or systemic complications) and preoperative treatment with erythropoietin were also recorded. RESULTS: Data for 166 patients were analyzed. Of these, 120 (72.3%) received tranexamic acid, 15 (9%) received epsilon-aminocaproic acid, and 31 (18.7%) received no antifibrinolytic treatment. Transfusions were given to 17 patients, of whom 6 (5.0%) had received tranexamic acid, 2 (133%) had received epsilon aminocaproic acid, and 9 (29.0%) had received no antifibrinolytic treatment. The mean numbers of packed red blood cell units transfused in each group were as follows: 0.075 in the tranexamic acid group, 0.200 in the epsilon-aminocaproic acid group, and 0.645 in the group with no antifibrinolytic treatment (P < .001). The mean decrease in hemoglobin levels 5 days after surgery was 3.04 g/dL in the tranexamic acid group, 3.55 g/dL in the epsilon-aminocaproic acid group and 3.76 g/dL in the group with no antifibrinolytic treatment (P < .001). CONCLUSIONS: Tranexamic acid is effective in reducing the percentage of patients requiring transfusions and in the number of units of packed red blood cells required in total knee replacement surgery. No complications attributable to this treatment were found. PMID- 18383969 TI - [Propofol-induced myocardial depression: possible role of atrial muscarinic cholinergic receptors]. AB - OBJECTIVE: To investigate the possible role of muscarinic cholinergic receptors (MCRs) in the depression of myocardial function induced by propofol, an intravenous anesthetic chemically unrelated to other drugs. Although adverse effects are rare, bradycardia has been reported and this can lead to cardiac arrest in some patients. The mechanism behind this effect is still unknown but a possible role for MCRs has been suggested. MATERIAL AND METHODS: The interaction of propofol with human atrial MCRs was determined by means of inhibition tests using [3H] quinuclidinyl benzilate ([3H] QNB). RESULTS: The displacement of [3H] QNB binding to human atrial MCRs by propofol was concentration dependent but the observed effect was not consistent with a model of simple competition between propofol and [3H] QNB. CONCLUSION: Propofol appears to have the ability to modify the activity of human atrial MCRs and this effect may be related to its ability to induce bradycardia. PMID- 18383970 TI - [Remifentanil bolus for cesarean section in high-risk patients: study of 12 cases]. AB - OBJECTIVES: To evaluate the utility and safety of remifentanil for hemodynamic control during cesarean section in high-risk patients ineligible for spinal anesthesia. METHODS: One minute before induction we injected a bolus of 1 microg x kg(-1) of remifentanil, followed by propofol (2.5 mg x kg(-1)), succinylcholine (1 mg x kg(-1)), cisatracurium, sevoflurane in oxygen and nitrous oxide, and fentanyl (5 microg x kg(-1)) after clamping the umbilical cord. We recorded maternal hemodynamic variables, pulse oximetry, capnography, bispectral index, and presence of muscular rigidity. In the neonate we assessed fetal wellbeing, weight, and requirement for naloxone. Hemodynamic stability was defined as no more than 15% variation in arterial pressure with respect to baseline. RESULTS: Twelve patients undergoing surgery because of placenta abruptio, subarachnoid hemorrhage, HELLP syndrome, or preeclampsia were enrolled. Hemodynamic variables were consistently stable during surgery in all patients. No cases of neonatal rigidity were noted and there was no need for naloxone. The mean Apgar score was 6.42 (1.5) at 1 minute and 8.42 (0.9) at 5 minutes. CONCLUSION: Bolus injection of 1 microg x kg(-1) of remifentanil may be useful for maintaining maternal hemodynamic stability in high-risk obstetric cases. Given the risk of neonatal depression, this resource should be used selectively and the means for neonatal resuscitation should be available. PMID- 18383971 TI - [Minimally invasive cardiopulmonary monitoring with the PiCCO Plus system]. AB - Insertion of a central venous catheter and an arterial catheter would be indicated in hemodynamically unstable or severely hypoxic patients in critical care units. In this setting, cardiorespiratory monitoring by transpulmonary thermodilution (TPTD) can be considered minimally invasive given that only a single arterial thermodilution catheter and a single central venous catheter are required to be connected to a specific monitor (the PiCCO Plus, Pulsion Medical Systems, Munich, Germany). TDTP simultaneously measures cardiac output, preloading, and cardiac function in hemodynamically unstable patients and predicts the response to volume. The technique can be managed by any health care professional. In hypoxic patients, TDTP identifies cases of pulmonary edema that might benefit from a negative fluid balance, evaluates pulmonary vascular permeability, facilitates our understanding of pathophysiologic mechanisms of hypoxemia, and predicts the likelihood of deleterious hemodynamic effects of positive end-expiratory pressures. PMID- 18383972 TI - [Supplemental oxygen for the prevention of postoperative nausea and vomiting: a meta-analysis of randomized clinical trials]. AB - OBJECTIVE: Despite the development of antiemetic drugs, the incidence of postoperative nausea and vomiting remains between 20% and 30%. This meta-analysis examines the hypothesis that perioperative administration of supplemental oxygen reduces the incidence of these complications. METHODS: We performed a systematic search of the literature through MEDLINE, EMBASE, the Cochrane Library, reference lists, and a manual search, with no language restrictions, up to September 2007 to identify randomized clinical trials evaluating the effect o f supplemental oxygen on postoperative nausea and vomiting. The data were extracted and analyzed using the RevMan program, version 4.2.9 (Cochrane Collaboration, Oxford, UK). RESULTS: The study included 9 randomized clinical trials with a total of 1661 enrolled patients (824 assigned to the group with a higher oxygen concentration and 837 assigned to the group with a lower oxygen concentration). Perioperative supplemental oxygen has no effect on the incidence of nausea (relative risk [RR], 0.94; 95% confidence interval [CI], 0.82 to 1.08), postoperative nausea and/or vomiting (RR, 0.93; 95% CI, 0.74 to 1.17), or the need for rescue antiemetic drugs (RR, 0.90; 95% CI, 0.70 to 1.15). The incidence of vomiting, however, is reduced (RR, 0.77; 95% CI, 0.62 to 0.97). Significant differences were not found in the incidence of atelectasis (RR, 1.23; 95% CI, 0.50 to 3.00) or postoperative PaCO2 (weighted mean difference, -4.0; 95% CI, -12.3 to 4.3). CONCLUSIONS: Supplemental oxygen reduces the incidence of postoperative vomiting. Administration of supplemental oxygen could be an effective method of reducing postoperative vomiting but does not replace current indications for pharmacologic prophylaxis. PMID- 18383973 TI - [Perioperative transesophageal echocardiography: proposal for a standardized report form]. PMID- 18383974 TI - [Cardiac perforation as a late complication in a man with an implantable cardioverter-defibrillator]. AB - A 55-year-old man with an implantable cardioverter-defibrillator (ICD) placed after diagnosis of Brugada syndrome 4 years earlier was admitted to hospital with dyspnea and a large left pleural effusion. After several episodes of cardiorespiratory arrest and application of advanced cardiac life support measures, an emergency sternotomy was performed. Cardiac and pleural perforation by the ICD lead was observed and the device was removed. Since the ICD was introduced in 1980, it has been effective in the treatment of malignant ventricular arrhythmias and in reducing the incidence of sudden death. Increased use, however, has meant a rise in the number of complications, some of which are potentially fatal. The rare complication we describe should therefore be considered whenever a patient with an ICD develops sudden respiratory failure or massive hemoptysis that cannot be explained by other causes. PMID- 18383975 TI - [Spinal cord compression by tumor with onset following epidural catheterization]. AB - We report the case of a 16-year-old boy with a pelvic tumor who developed signs of spinal cord compression following placement of a lumbar epidural catheter to obtain a percutaneous biopsy. The fact that a catheter had been inserted led us to suspect epidural hematoma as a complication, but surgery revealed that the pelvic tumor had spread to the lumbar epidural space and was causing motor blockade of both legs. PMID- 18383976 TI - [Anesthetic considerations in nemaline myopathy]. PMID- 18383977 TI - [Convulsions after a test dose of ropivacaine and negative aspiration test for combined spinal-epidural analgesia for cesarean section]. PMID- 18383978 TI - [Paravertebral block for major breast surgery in a patient with amyotrophic lateral sclerosis]. PMID- 18383979 TI - [Rapid preoperative preparation with iopanoic acid in severe liver toxicity due to methimazole: anesthetic implications]. PMID- 18383980 TI - [Clinical utility of the bispectral index for detecting epileptic seizures]. PMID- 18383981 TI - [Noninvasive ventilation through a helmet following facial reconstruction surgery]. PMID- 18383982 TI - [Anterior giant cervical osteophyte as a possible cause of difficult intubation]. PMID- 18383983 TI - [Difficult airway in a patient with a giant hemangioma of the tongue]. PMID- 18383984 TI - Does dementia exist? Dispelling the myth. PMID- 18383985 TI - Does complementary and alternative medicine represent only placebo therapies? PMID- 18383986 TI - Progress notes updated: the consortium and other developments in education in complementary and integrative medicine. PMID- 18383987 TI - Efficacy of lifestyle changes in modifying practical markers of wellness and aging. AB - PURPOSE: To determine the efficacy of asking people to add fiber, exercise, and stress management to their lifestyles to enhance markers of wellness and aging. METHODS: A 10-week, randomized control study conducted in a wellness center in St Petersburg, Florida. Participants were adults aged 21 to 65 years who exercised fewer than 3 days per week. Fifty-six subjects were randomized to a control or an intervention group. Subjects followed a diet with > 30 g of fiber and < 16 g of saturated fat daily and were taught to reach 70% to 85% of their maximum heart rate 5 to 6 days per week and to perform strength training 3 days per week. They were also asked to participate in 10 to 20 minutes of stress management activities daily. The study was designed to determine changes in body composition, maximal rate of oxygen consumption (VO2 max), total cholesterol: high-density lipoprotein (TC:HDL) ratio, and cognition. RESULTS: Initial analyses with analysis of variance (ANOVA) comparing the intervention group to the control group showed significant improvements in TC:HDL (8.9% average; P = .02) and change in weight (2.3 kg average; P = .016). When the groups were compared, the improvements in cognitive flexibility and VO2 max with ANOVA were not significant (P = .17 and P = .11, respectively). Additional independent t tests showed decreases in TC:HDL of 8.9% (P = .02) and TC of 7.3% (P = .001) for the intervention group compared to the control group. A mean increase of 29% in VO2 max of intervention subjects who exercised aerobically for at least 30 minutes 5 days/week was significant (P = .02) compared to the control group. Over the 10 weeks, the control group showed no significant change in lipids, body composition, cognition, and fitness, whereas the intervention group showed decreased body mass index (BMI) of 0.72 (P = .02), weight loss of 2.3 kg (P = .016), and decreased body fat of 1.6% (P < .0001). In the intervention group, those with a BMI > 24 who exercised 5 to 6 days/week lost 4.8 kg and 4.1 kg in body fat. Also, in the intervention group, several of the cognitive scores showed statistically significant improvements from baseline: mental speed (4.6%, P = .014), reaction time (4.5%, P = .023), and cognitive flexibility (11.7%, P = .019), but none of these cognitive changes was significant with independent t testing when compared to the control group. CONCLUSIONS: A diet high in fiber and low in saturated fat combined with strength training, aerobic activity, and stress management activities improves fitness and several markers of wellness and aging. PMID- 18383988 TI - Cost-effectiveness of naturopathic care for chronic low back pain. AB - OBJECTIVE: To determine the cost-effectiveness of naturopathic care (acupuncture, relaxation exercises, exercise and dietary advice, and a back care booklet) compared to standardized physiotherapy education and a back care booklet (control treatment) for low back pain in a sample (N = 70) of warehouse workers. DESIGN: Economic evaluation based upon the results of a pragmatic randomized controlled trial to determine the cost-effectiveness of naturopathic care to society as a whole, to the employer, and to participants. RESULTS: Naturopathic care (as compared to the control treatment) significantly improved quality-adjusted life years over the 6-month study period (3-month intervention period and 3-month follow-up period) by 0.0256 (95% CI: 0.0075, 0.0437)--roughly equivalent to 9.4 "perfect health" days. Naturopathic care also significantly reduced societal costs by $1212 per participant. From the perspective of the employer, the intervention cost $154 per absentee day avoided (compared to employer costs of lost productivity of $172 per day) and had a return on investment of 7.9% under the healthcare coverage limits set by this employer and assuming the employer paid the full cost of naturopathic care. Participants experienced savings in adjunctive care of $1096 per participant. CONCLUSIONS: This economic evaluation alongside a pragmatic randomized control trial shows naturopathic care to be more cost-effective than a standardized physiotherapy education regimen in the treatment of chronic low back pain. Further studies of the economic impact of naturopathic medicine are warranted. PMID- 18383989 TI - Discerning the Mauve Factor, Part 1. AB - "Mauve Factor" was once mistaken for kryptopyrrole but is the hydroxylactam of hemopyrrole, hydroxyhemopyrrolin-2-one (HPL). Treatment with nutrients- particularly vitamin B6 and zinc--reduces urinary excretion of HPL and improves diverse neurobehavioral symptoms in subjects with elevated urinary HPL. Heightened HPL excretion classically associates with emotional stress, which in turn is known to associate with oxidative stress. For this review, markers for nutritional status and for oxidative stress were examined in relationship to urinary HPL. In cohorts with mixed diagnoses, 24-hour urinary HPL correlated negatively with vitamin B6 activity and zinc concentration in red cells (P < .0001). Above-normal HPL excretion corresponded to subnormal vitamin B6 activity and subnormal zinc with remarkable consistency. HPL correlated inversely with plasma glutathione and red-cell catalase, and correlated directly with plasma nitric oxide (P < .0001). Thus, besides implying proportionate needs for vitamin B6 and zinc, HPL is a promising biomarker for oxidative stress. HPL is known to cause non-erythroid heme depression, which lowers zinc, increases nitric oxide, and increases oxidative stress. Administration of prednisone reportedly provoked HPL excretion in animals. Since adrenocorticoid (and catecholamine) stress hormones mediate intestinal permeability, urinary HPL examined in relationship to urinary indicans, presumptive marker for intestinal permeability. Urinary HPL associated with higher levels of indicans (P < .0001). Antibiotics reportedly reduce HPL in urine, suggesting an enterobic role in production. Potentially, gut is a reservoir for HPL or its precursor, and stress-related changes in intestinal permeability mediate systemic and urinary concentrations. PMID- 18383990 TI - Increasing research capacity at the New England School of Acupuncture through faculty and student research training initiatives. AB - Few complementary and alternative medicine (CAM) institutions require their students to undergo substantive training in research literacy and conduct, and well-developed programs to train CAM institution faculty in research are virtually non-existent. As part of a National Center for Complementary and Alternative Medicine (NCCAM) initiative to increase research capacity at CAM institutions, the New England School of Acupuncture (NESA), in collaboration with the Harvard Medical School (HMS) Osher Institute, was awarded a Developmental Center for Research on Complementary and Alternative Medicine (DCRC) grant. This article discusses a number of initiatives that we designed and implemented to train NESA students, faculty members, and alumni in the foundations of clinical research and to stimulate interest in both participating in research and receiving additional research training. Specific initiatives included a 30-hour faculty "Foundations of Research" course; a year-long course entitled, "How to Write a Publishable Case Report"; institution of a monthly research seminar series; revision of an already required student research course; and the addition of 2 new student-mentored independent research electives. We discuss successes and challenges encountered in developing and administering these initiatives and the overall impact they have had on research culture and productivity at NESA. PMID- 18383991 TI - Randomized controlled trials as evidence in legal disputes about the benefits of complementary and alternative medicine. PMID- 18383992 TI - Jacob Liberman, OD, PhD: change your vision, change your life. Interview by Marc David and Suzanne Snyder. PMID- 18383993 TI - Hepatitis C in India--unanswered questions. PMID- 18383994 TI - Review of pathogenesis and management of constipation. AB - This article reviews the pathogenesis, classification, mechanism and management of constipation. Constipation is likely to be common in the Indian population. It is difficult to define precisely since perception of patient and doctor may differ. Rome Consensus Criteria may not be applicable in India where we should not define constipation as stool frequency less than thrice a week as normal bowel movement in among Indians is different than that in the West. Constipation may be due to difficulty in evacuation, i.e. dyschezia, or due to a combination of infrequency and dyschezia. Low fibre diet, insufficient fluid intake, irregular toilet habit, lack of exercise, prolonged bed rest and chronic consumption of drugs may all lead to this chronic ailment. Constipation may result from slow colonic transit, faecal evacuation disorders or a combination of both. The first step in management is to exclude organic and anatomic causes. In the elderly, proctosigmoidoscopy or when required, colonoscopy and barium enema should be done. Colonic transit study is useful to screen for slow transit constipation or faecal evacuation disorders. Defecography, the balloon expulsion test, anorectal ultrasound, anorectal manometry, defecometry, anal sphincter electromyography and the pudendal nerve terminal motor latency study may be used to diagnose faecal evacuation disorders. Treatment aims at symptom relief and bettering the quality of life. High fibre diet, physical activity, modification of current therapy (e.g. where the patient is on opioids), and prescription of laxatives may provide relief. Current guidelines for prescribing laxatives suggest bulk agents as first line and osmotic agents as second line therapy. Biofeedback is useful in faecal evacuation disorders. Surgery may also rarely be necessary to correct anatomical abnormalities. PMID- 18383995 TI - Living donor liver transplantation in India. AB - The world over, liver transplantation has emerged a panacea for thousands of patients suffering from end-stage liver disease. The strides made in living donor liver transplantation (LDLT) by Asian centres particularly in Japan, Korea, Hong Kong and Taiwan made many Indian centres realise that in order to sustain liver transplant activity in the country, a similar solution had to be found. Even though LDLT is very resource intensive and requires skilled multidisciplinary manpower, 22 centres in India have performed liver transplants, of which 14 have performed at least one LDLT procedure. 140 LDLT procedures have been performed at our centre, of which 13 have been done in emergency circumstances. LDLT has certain advantages over DDLT. It allows for adequate preparation of the patient for elective transplant and recipients are not in competition with others over the same donor organ. Major concerns with LDLT are of donor safety and biliary complications. In conclusion, establishing a high volume LDLT centre with excellent success rates is feasible in the Indian setting. PMID- 18383996 TI - Do different hepatitis C virus genotypes behave differently? AB - BACKGROUND: The natural history of hepatitis C genotype III infection, the predominant form in India, is not wholly understood. This study attempted to compare the natural history of diseases due to genotypes III and I. METHODS: This 10-year prospective follow-up study (mean follow-up period = 3.6 +/- 1.4, range = 1-10 years) included 108 patients of hepatitis C. Group 1 comprised 65 patients with hepatitis C genotype III infection (mean age = 46.1 +/- 11.3 years, male: female = 1.8 : 1) and group 2 comprised 43 patients with hepatitis C genotype I infection (mean age = 44.2 +/- 8.2 years, male: female = 2.1 : 1). Demographic features, clinical presentation and course, response to treatment (either interferon-ribavirin or peginterferon-ribavirin combination) and complications were noted for all patients. Data were analysed using the chi-square test and Student's t-test. RESULTS: The number of steatosis cases was larger in group 1 (32.3%, 21/65 patients) than in group 2 (18.6%, 8/43 patients) although statistically not significant. There was no significant difference in the mode of infection, presence of diabetes, obesity or alcoholism, clinical presentation, extra-hepatic manifestations, stage of liver disease, complications like decompensation or hepatocellular carcinoma and mortality. Overall, the sustained treatment response was significantly greater in group 1 patients [(87.5%, 21/24 treated patients vs. 56.2%, 9/16 treated patients in group 2; p = 0.0001)]. CONCLUSION: HCV with genotype III was associated with better treatment response. Although statistically not significant, more number of patients in genotype III had steatosis. PMID- 18383997 TI - Spectrum and outcome of pancreatic trauma. AB - BACKGROUND AND AIMS: Pancreatic trauma is associated with high morbidity and mortality. Diagnosis is often difficult and surgery poses a formidable challenge. METHOD: Data from 17 patients of pancreatic trauma gathered from a prospectively maintained database were analysed and the following parameters were considered: mode of injury, diagnostic modalities, associated injury, grade of pancreatic trauma and management. Pancreatic trauma was graded from I through IV, as per Modified Lucas Classification. RESULTS: The median age was 39 years (range 19 61). The aetiology of pancreatic trauma was blunt abdominal trauma in 14 patients and penetrating injury in 3. Associated bowel injury was present in 4 cases (3 penetrating injury and 1 blunt trauma) and 1 case had associated vascular injury. 5 patients had grade I, 3 had grade II, 7 had grade III and 2 had grade IV pancreatic trauma. Contrast enhanced computed tomography scan was used to diagnose pancreatic trauma in all patients with blunt abdominal injury. Immediate diagnosis could be reached in only 4 (28.5%) patients. 7 patients responded to conservative treatment. Of the 10 patients who underwent surgery, 6 required it for the pancreas and the duodenum. (distal pancreatectomy with splenectomy-3, pylorus preserving pancreatoduodenectomy-1, debridement with external drainage-1, associated injuries-duodenum-1). Pancreatic fistula, recurrent pancreatitis and pseudocyst formation were seen in 3 (17.05%), 2 (11.7%) and 1 (5.4%) patient respectively. Death occurred in 4 cases (23.5%), 2 each in grades III and IV pancreatic trauma. CONCLUSIONS: Contrast enhanced computed tomography scan is a useful modality for diagnosing, grading and following up patients with pancreatic trauma. Although a majority of cases with pancreatic trauma respond to conservative treatment, patients with penetrating trauma, and associated bowel injury and higher grade pancreatic trauma require surgical intervention and are also associated with higher morbidity and mortality. PMID- 18383998 TI - Efficacy and tolerability of pegylated interferon alpha 2b and ribavirin in chronic hepatitis C--a report from eastern India. AB - AIM: To study the efficacy and tolerability of pegylated interferon alpha 2b and ribavirin therapy in a cohort of chronic hepatitis C patients. METHODS: In a prospective, open label, uncontrolled trial pegylated interferon alpha 2b (Viraferon Peg) 1.5 microgram/ kg subcutaneously weekly plus daily ribavirin 800mg for 24 weeks in genotypes 2 & 3 and 1000mg for 48 weeks in genotypes 1 and 4 was administered to 16 patients of chronic hepatitis C. The primary end point was the sustained viral response. Therapy was prolonged by 3 months if the end of therapy response was not attained. Drug dosage was modified or temporarily discontinued if anaemia or bone marrow suppression developed. RESULTS: Both virological end of therapy response and sustained viral response were seen in 75% cases but not every patient who achieved end of therapy response had a sustained viral response. Relapse was seen in 31% cases and a pattern of delayed response was seen in 2 patients who later experienced a sustained viral response. Biochemical and virological responses were similar. A lower baseline viral load, genotype 3, a high ALT and the parenteral mode of viral acquisition were associated with higher sustained viral response rates. A good response was also seen in men, those over 50 years of age and those with normal baseline ALT. Most relapses occurred in genotype 3 patients whose age was less than 50 years; however the relapsing viral load was very low. 66% of previous interferon and ribavirin non-responders achieved sustained viral response. Treatment was well tolerated; temporary dose modification was required in 3 patients. CONCLUSION: In Indian patients, a combination of peginterferon alpha 2b and ribavirin is safe and effective both as initial treatment of chronic hepatitis C and for use in previous non-responders. PMID- 18383999 TI - Time trends and pathological profile of carcinoma lower oesophagus and gastro oesophageal junction over the last 20 years--an experience from South India. AB - There is an upward trend in the incidence of adenocarcinoma lower oesophagus in western countries. However there is only limited comparable data from Asian countries. We conducted a retrospective analysis of our data compiled over a twenty-year period (1985-2004). All lesions diagnosed as either squamous cell carcinoma or adenocarcinoma involving the oesophagus with or without involvement of the gastro-oesophageal junction were included in the study. 476 cases with biopsy proven malignancy (either adeno or squamous) of lower oesophagus were studied. The pattern of change in frequency and histology over twenty years was analysed using the chi square test for trend. There was a consistent increase in the frequency of cancer involving the gastro-oesophageal junction though it did not reach statistical significance (p = 0.15). Out of 476 lower oesophageal cancers, 249 were adeno-carcinomas and 227 were squamous cell carcinomas. Adenocarcinoma involving the gastro-oesophageal junction showed consistent increase even though the p value was not significant (p = 0.09) and this therefore requires further longitudinal studies. There was no change in trend for pattern and frequency of squamous cell carcinoma oesophagus involving different sub-sites during the study period. PMID- 18384000 TI - Peritonitis: 10 years' experience in a single surgical unit. AB - BACKGROUND: Peritonitis secondary to gut perforation is still one of the commonest surgical emergencies in India and is associated with high morbidity and mortality. The present study examines the aetiology and outcome of peritonitis cases operated on in our surgical unit, and compares our findings with those of previous studies performed between 1981 and 1991. METHOD: A retrospective study of 260 peritonitis patients operated on in a single surgical unit from 1995 to 2006 was done and data involving clinical presentation, operative findings and post-operative course were studied and analysed. RESULTS: Causes of peritonitis were small bowel perforation (96 ileal, 17 jejunal), peptic perforation (45 duodenal, 16 gastric), appendicular perforation (36), primary peritonitis (8), and others (42). The incidence of major complications was 25% (burst-11%, leak 5%, intraabdominal abscess-5%, multi-organ failure-6.5%). The overall mortality was 10%. High mortality was observed in jejunal, gall bladder and liver abscess perforation cases (> 20%). Histopathological evaluation (143 specimens) revealed tuberculosis in 42 (mostly small bowel), malignancy in 8, and inflammation in the rest. Comparisons with a similar study carried out in the same unit and published in 1995 revealed similar demographic features and mortality, but a change in the most common cause (peptic ulcer perforation to small bowel perforation), and an increased performance of enterostomy compared with primary repair in small bowel perforation and a decrease in the leak rate (13% to 4%). CONCLUSION: Small bowel perforation is the commonest form of perforation and the mortality rate associated with peritonitis remains unchanged. PMID- 18384001 TI - Spectrum of microscopic colitis in a tertiary care centre in India. AB - INTRODUCTION: The incidence of microscopic colitis has recently increased. Although collagenous colitis and lymphocytic colitis are the two main subtypes of microscopic colitis, many patients may not fit into either category and are thus included under the header nonspecific colitis. Of late, the spectrum of microscopic colitis has widened to include minimal change colitis, microscopic colitis not otherwise specified and microscopic colitis with giant cells. There is a lack of information concerning the spectrum of microscopic colitis in Asia. METHOD: In a retrospective analysis, case records of 29 patients diagnosed with microscopic colitis between 1999-2005 were analysed. Drug use parasitic infection and common bacterial infections were excluded. Colonoscopic/ sigmoidoscopic examination was done and multiple colonic mucosal biopsies were stained serially with haematoxylin and eosin for detailed histological examination and Masson trichrome for sub-epithelial collagen band. Based on histological criteria, patients were categorised into five subtypes: collagenous colitis (presence of collagenous thickening of surface epithelium basement membrane > 10 microm), lymphocytic colitis (intra-epithelial lymphocytes more than 20 per 100 colonocytes), minimal change colitis (crypt architectural abnormality in the form of cryptitis and crypt dilatation in the absence of increase in intraepithelial lymphocytes and subepithelial collagenous band), microscopic colitis not otherwise specified (increased inflammatory cell infiltrates in the lamina propria in the absence of other abnormalities) and microscopic colitis with giant cells. RESULTS: Mean age of patients was 38.59 years (range 12-62). Of 29 patients with microscopic colitis, 7 (24.1%), 4 (13.8%), 7 (24.1%) and 11 (37.9%) were classified as collagenous colitis, lymphocytic colitis, minimal change colitis and microscopic colitis not otherwise specified, respectively. None of these patients had giant cells. There was no significant correlation between disease type and clinical manifestations. CONCLUSION: Microscopic colitis has a wide histological spectrum. Cases reported as non-specific colitis, may be categorised into definite subtypes of microscopic colitis. PMID- 18384002 TI - Mucormycosis of the bile duct: a case report. AB - A case of isolated localised mucormycosis of the bile duct in an immuno-competent 54 year old female patient is described. Mucormycosis is rare in immuno-competent patients. Isolated localised mucormycosis of the bile duct has not yet been described in the literature. PMID- 18384003 TI - External resection of a giant sigmoid lipoma causing colonic intussusception and prolapse through the anal canal. AB - We depict the case of an 80-year-old female patient who presented to us with a history of protruding mass per anum. Sigmoidoscopy revealed a large globular pedunculated polyp at 22 cm from the anal verge resulting in a sigmoidorectal intussusception. Endoscopic polypectomy was not technically possible due to the large size of the polyp. At the time of prolapse the polyp was tied at its pedicle with thread and resected surgically. The patient is asymptomatic on follow-up. PMID- 18384004 TI - Non-familial juvenile polyposis in prolapsed rectum. AB - Juvenile polyposis located solely on prolapsed rectal mucosa is very unusual. We report the case of a 17-year old boy who presented to us with a history of passage of blood and mucus per rectum of a mass protruding through the anus during defecation. Per rectal and colonoscopic examinations revealed numerous polyps located solely on the prolapsed rectal mucosa. Histopathology was consistent with juvenile polyposis. He was managed with repeated sessions of endoscopic polypectomy. Family screening was negative for colonic polyps. PMID- 18384005 TI - Oesophageal leiomyoma: case report and review of literature. PMID- 18384006 TI - Left gastric artery pseudoaneurysm following traumatic pancreatic transection. AB - A 24-year-old man presented to us 10 days after suffering blunt trauma to the abdomen. He was diagnosed with pancreatic transection and underwent distal pancreatectomy and splenectomy. Two weeks after the operation, he developed intra abdominal haemorrhage. Selective visceral angiogram revealed left gastric artery pseudoaneurysm, which had embolised. His recovery was uneventful. To our knowledge, ruptured left gastric artery pseudoaneurysm following pancreatic trauma, has not been reported before. In this article, we discuss some vascular complications of pancreatic trauma. PMID- 18384007 TI - Aetiology, clinical profile and prognostic indicators for children with acute liver failure admitted in a teaching hospital in Kolkata. AB - OBJECTIVE: To estimate the prevalence of hepatotropic viruses in the causation of acute liver failure in children admitted to a tertiary hospital in Kolkata. DESIGN: Analysis of clinical and laboratory parameters (including viral markers) of children with acute liver failure using a predesigned, structured proforma. SUBJECTS AND SETTINGS: Admitted patients aged from 1 through 12 years who met the criteria of acute liver failure were included in the study. RESULTS: Of the 45 patients in our study, a majority was from the southern part of West Bengal. Their mean age was 7.12 +/- 0.37 years. The male: female ratio was 1:1.25. It was possible to determine the aetiology in 35 of the 45 patients (77.7%) admitted. Of these 35, a diagnosis of hepatitis due to hepatotropic viruses was made in 30 patients. The hepatitis A virus was responsible for 16 of the 30 cases (53.3%), 9 cases attributed to HAV only. Following this was the hepatitis E virus causing ALF in 14 cases (46.6%), 7 singularly so. Hepatitis B virus caused 8 cases (26.6%), 6 singly. The survival rate during hospital stay was 51.1%. Prodrome, decreased liver span, ascites, cerebral oedema, coagulopathy, renal failure, spontaneous bacterial peritonitis, signs and symptoms of clinical sepsis (corroborated by laboratory data), severe hypoalbuminaemia and electrolyte imbalance were significantly more in patients who died. The mean age, prothrombin time, serum bilirubin level and stage of encephalopathy differed significantly between survivors and non-survivors. CONCLUSION: In southern Bengal, hepatotropic viruses are the predominant cause of acute liver failure in children. Of these, the hepatitis viruses A and E transmitted via the enteric route dominate (24 of 30 cases). PMID- 18384008 TI - Gut flora in health and small intestinal bacterial overgrowth syndrome. PMID- 18384009 TI - [Natural maternal behavior in the swine and their expression in modern swine husbandry: a literature review]. PMID- 18384010 TI - [Drug resistance in Salmonella; comparison of isolates from Vietnam and the Netherlands]. PMID- 18384011 TI - [Flea circus]. PMID- 18384012 TI - [FELV infection, the kiss of death]. PMID- 18384013 TI - [Occupational dermatitis in veterinarians]. PMID- 18384014 TI - [Instruments, books and other objects memorable to (almost) forgotten opinions, therapies, buildings, et cetera. Cat holder]. PMID- 18384015 TI - [Applying HACCP-based quality risk management on dairy farms]. PMID- 18384016 TI - [Good health care guaranteed]. PMID- 18384017 TI - [Deworming medicine and prescription mandatory for horses]. PMID- 18384018 TI - [EVDS- Congress about veterinary dentistry]. PMID- 18384019 TI - Electrokinetic analyte transport assay for alpha-fetoprotein immunoassay integrates mixing, reaction and separation on-chip. AB - A rapid and highly sensitive CE immunoassay method integrating mixing, reaction, separation, and detection on-chip is described for the measurement of alpha fetoprotein (AFP), a liver cancer marker in blood. Antibody-binding reagents, consisting of 245-bp DNA coupled anti-AFP WA1 antibody (DNA-WA1) and HiLyte dye labeled anti-AFP WA2 antibody (HiLyte-WA2), and AFP-containing sample were filled into adjacent zones of a chip channel defined by the laminar flow lines of the microfluidic device using pressure-driven flow. The channel geometry was thus used to quantitatively aliquot the reagents and sample into the chip. DNA-WA1 was electrokinetically concentrated in the channel and sequentially transported through the AFP-sample zone and HiLyte-WA2 zone by ITP in such a manner that the AFP sandwich immune complex formation took place in the sample and HiLyte-WA2 zones. The sandwich AFP immune complex was then detected by LIF after CGE in a separation channel that was arranged downstream of the reaction channel. AFP was detected within 136 s with a detection sensitivity of 5 pM. The on-chip immunoassay described here, applying ITP concentration, in-channel reaction, and CGE separation, has the potential of providing a rapid and sensitive method for both clinical and research applications. PMID- 18384020 TI - High-throughput kinetic study of hydrogenation over palladium nanoparticles: combination of reaction and analysis. AB - The hydrogenation of 1-acetylcyclohexene, cyclohex-2-enone, nitrobenzene, and trans-methylpent-3-enoate catalyzed by highly active palladium nanoparticles was studied by high-throughput on-column reaction gas chromatography. In these experiments, catalysis and separation of educts and products is integrated by the use of a catalytically active gas chromatographic stationary phase, which allows reaction rate measurements to be efficiently performed by employing reactant libraries. Palladium nanoparticles embedded in a stabilizing polysiloxane matrix serve as catalyst and selective chromatographic stationary phase for these multiphase reactions (gas-liquid-solid) and are coated in fused-silica capillaries (inner diameter 250 microm) as a thin film of thickness 250 nm. The palladium nanoparticles were prepared by reduction of palladium acetate with hydridomethylsiloxane-dimethylsiloxane copolymer and self-catalyzed hydrosilylation with methylvinylsiloxane-dimethylsiloxane copolymer to obtain a stabilizing matrix. Diphenylsiloxane-dimethylsiloxane copolymer (GE SE 52) was added to improve film stability over a wide range of compositions. Herein, we show by systematic TEM investigations that the size and morphology (crystalline or amorphous) of the nanoparticles strongly depends on the ratio of the stabilizing polysiloxanes, the conditions to immobilize the stationary phase on the surface of the fused-silica capillary, and the loading of the palladium precursor. Furthermore, hydrogenations were performed with these catalytically active stationary phases between 60 and 100 degrees C at various contact times to determine the temperature-dependent reaction rate constants and to obtain activation parameters and diffusion coefficients. PMID- 18384021 TI - Continuous and precise particle separation by electroosmotic flow control in microfluidic devices. AB - A new scheme has been described for continuous particle separation using EOF in microfluidic devices. We have previously reported a method for particle separation, called "pinched flow fractionation (PFF)", in which size-dependent and continuous particle separation can be achieved by introducing pressure-driven flows with and without particles into a pinched microchannel. In this study, EOF was employed to transport fluid flows inside a microchannel. By controlling the applied voltage to electrodes inserted in each inlet/outlet port, the flow rates from both inlets, and flow rates distributed to each outlet could be accurately tuned, thus enabling more effective separation compared to the pressure-driven scheme. In the experiment, the particle behaviors were compared between EOF and pressure-driven flow schemes. In addition, micrometer- and submicrometer-sized particles were accurately separated and individually collected using a microchannel with multiple outlet branch channels, demonstrating the high efficiency of the presented scheme. PMID- 18384022 TI - Synthesis, characterization, and field-effect transistor properties of carbazolenevinylene oligomers: from linear to cyclic architectures. AB - Two cyclic carbazolenevinylene dimers 1 and 2 were synthesized by McMurry coupling reactions. A linear compound 3 was also prepared for comparison. Compounds 1-3 were fully characterized by means of NMR spectroscopy, HRMS, elemental analysis, and UV/Vis absorption spectroscopy. Quantum chemical simulations showed that the cyclic compounds possessed smaller HOMO-LUMO gaps and more extended conjugation. The UV/Vis absorption spectra of the cyclic compounds showed blueshifts compared with that of the linear compound 3. Time-dependent DFT (TD-DFT) analysis revealed that this was due to the different selection rules for molecules with cyclic and linear architectures. The cyclic conformation also significantly affected the molecular ordering in the solid state. The X-ray crystal structure of 1 showed partial pi-pi overlapping between the adjacent molecules. Thin films of 1-3 were fabricated by the vacuum-deposition method on Si/SiO(2) substrates. Multicrystalline thin films were obtained from compounds 1 and 2, but only amorphous thin films could be obtained for the linear compound 3. Another important difference between the cyclic and linear compounds was the reduced reorganization energy for the cyclic compounds. These two facts have resulted in improved field-effect transistor (FET) mobilities for the cyclic compounds compared with the linear compound. In addition, as the substrate temperature has a significant influence on the morphology and the degree of crystallinity of the thin films deposited, the device performance could be optimized by varying the substrate temperature. The FET devices based on 2 gave the highest mobility of 0.013 cm(2) V(-1) s(-1). The results showed that carbazole derivatives with cyclic structures might make better FETs. PMID- 18384023 TI - New strategies for the synthesis of pyrimidine derivatives. AB - Recent advances in pyrimidine synthesis are described. Modification of conventional strategies involving N-C-N fragment condensation with 1,3-dicarbonyl derivatives remains a common theme in current literature. Other methods, including N-C fragment condensation strategies, provide reactive intermediates capable of intramolecular cyclization and formation of pyrimidine derivatives. These recently developed methodologies offer a valuable addendum to azaheterocycle synthesis. PMID- 18384024 TI - Serine hydroxymethyl transferase from Streptococcus thermophilus and L-threonine aldolase from Escherichia coli as stereocomplementary biocatalysts for the synthesis of beta-hydroxy-alpha,omega-diamino acid derivatives. AB - A novel serine hydroxymethyl transferase from Streptococcus thermophilus (SHMT) and a L-threonine aldolase from Escherichia coli (LTA) were used as stereocomplementary biocatalysts for the aldol addition of glycine to N-Cbz amino aldehydes and benzyloxyacetaldehyde (Cbz=benzyloxycarbonyl). Both threonine aldolases were classified as low-specific L-allo-threonine aldolases, and by manipulating reaction parameters, such as temperature, glycine concentration, and reaction media, SHMT yielded exclusively L-erythro diastereomers in 34-60 % conversion, whereas LTA gave L-threo diastereomers in 30:70 to 16:84 diastereomeric ratios and with 40-68 % conversion to product. SHMT is among the most stereoselective L-threonine aldolases described. This is due, among other things, to its activity-temperature dependence: at 4 degrees C SHMT has high synthetic activity but negligible retroaldol activity on L-threonine. Thus, the kinetic L-erythro isomer was largely favored and the reactions were virtually irreversible, highly stereoselective, and in turn, gave excellent conversion. It was also found that treatment of the prepared N-Cbz-gamma-amino-beta-hydroxy alpha-amino acid derivatives with potassium hydroxide (1 m) resulted in the spontaneous formation of 2-oxazolidinone derivatives of the beta-hydroxyl and gamma-amino groups in quantitative yield. This reaction might be useful for further chemical manipulations of the products. PMID- 18384025 TI - A one-pot synthesis of constitutionally unsymmetrical rotaxanes using sequential Cu(I)-catalyzed azide-alkyne cycloadditions. AB - A one-pot sequential Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) strategy is presented for the synthesis of constitutionally unsymmetrical cyclobis(paraquat-p-phenylene)-based rotaxanes in good yields from simple starting materials. The methodology consists of performing multiple CuAAC reactions to stopper a pseudorotaxane in a stepwise manner, the order of which is controlled through silyl-protection and Ag(I)-catalyzed deprotection of a terminal alkyne. The methodology is highlighted by the synthesis of an amphiphilic branched [4]rotaxane. The methodology increases the ability to access ever more complicated mechanically interlocked compounds to serve in devices as sophisticated and functional molecular machinery. PMID- 18384026 TI - New highly asymmetric Henry reaction catalyzed by Cu(II) and a C(1)-symmetric aminopyridine ligand, and its application to the synthesis of miconazole. AB - A new catalytic asymmetric Henry reaction has been developed that uses a C(1) symmetric chiral aminopyridine ligand derived from camphor and picolylamine. A variety of aromatic, heteroaromatic, aliphatic, and unsaturated aldehydes react with nitromethane and other nitroalkanes in the presence of DIPEA (1.0 equiv), Cu(OAc)(2)*H(2)O (5 mol %), and an aminopyridine ligand (5 mol %) to give the expected products in high yields (up to 99 %), moderate-to-good diastereoselectivites (up to 82:18), and excellent enantioselectivities (up to 98 %). The reaction is air-tolerant and has been used in the synthesis of the antifungal agent miconazole. PMID- 18384027 TI - A general method for the rapid synthesis of hollow metallic or bimetallic nanoelectrocatalysts with urchinlike morphology. AB - We have reported a facile and general method for the rapid synthesis of hollow nanostructures with urchinlike morphology. In-situ produced Ag nanoparticles can be used as sacrificial templates to rapidly synthesize diverse hollow urchinlike metallic or bimetallic (such as Au/Pt) nanostructures. It has been found that heating the solution at 100 degrees C during the galvanic replacement is very necessary for obtaining urchinlike nanostructures. Through changing the molar ratios of Ag to Pt, the wall thickness of hollow nanospheres can be easily controlled; through changing the diameter of Ag nanoparticles, the size of cavity of hollow nanospheres can be facilely controlled; through changing the morphologies of Ag nanostructures from nanoparticle to nanowire, hollow Pt nanotubes can be easily designed. This one-pot approach can be extended to synthesize other hollow nanospheres such as Pd, Pd/Pt, Au/Pd, and Au/Pt. The features of this technique are that it is facile, quick, economical, and versatile. Most importantly, the hollow bimetallic nanospheres (Au/Pt and Pd/Pt) obtained here exhibit an area of greater electrochemical activity than other Pt hollow or solid nanospheres. In addition, the approximately 6 nm hollow urchinlike Pt nanospheres can achieve a potential of up to 0.57 V for oxygen reduction, which is about 200 mV more positive than that obtained by using a approximately 6 nm Pt nanoparticle modified glassy carbon (GC) electrode. Rotating ring-disk electrode (RRDE) voltammetry demonstrates that approximately 6 nm hollow Pt nanospheres can catalyze an almost four-electron reduction of O(2) to H(2)O in air-saturated H(2)SO(4) (0.5 M). Finally, compared to the approximately 6 nm Pt nanoparticle catalyst, the approximately 6 nm hollow urchinlike Pt nanosphere catalyst exhibits a superior electrocatalytic activity toward the methanol oxidation reaction at the same Pt loadings. PMID- 18384028 TI - Role of the NH2 functionality and solvent in terdentate CNN alkoxide ruthenium complexes for the fast transfer hydrogenation of ketones in 2-propanol. AB - The reaction of [RuCl(CNN)(dppb)] (1; HCNN=6-(4-methylphenyl)-2 pyridylmethylamine) with NaOiPr in 2-propanol/C6D6 affords the alcohol adduct alkoxide [Ru(OiPr)(CNN)(dppb)].n iPrOH (5), containing the Ru-NH2 linkage. The alkoxide [Ru(OiPr)(CNN)(dppb)] (4) is formed by treatment of the hydride [Ru(H)(CNN)(dppb)] (2) with acetone in C6D6. Complex 5 in 2-propanol/C6D6 equilibrates quickly with hydride 2 and acetone with an exchange rate of (5.4+/ 0.2) s(-1) at 25 degrees C, higher than that found between 4 and 2 ((2.9+/-0.4) s(-1)). This fast process, involving a beta-hydrogen elimination versus ketone insertion into the Ru-H bond, occurs within a hydrogen-bonding network favored by the Ru-NH2 motif. The cationic alcohol complex [Ru(CNN)(dppb)(iPrOH)](BAr(f)4) (6; Ar(f)=3,5-C6H3(CF3)2), obtained from 1, Na[BAr(f)4], and 2-propanol, reacts with NaOiPr to afford 5. Complex 5 reacts with either 4,4'-difluorobenzophenone through hydride 2 or with 4,4'-difluorobenzhydrol through protonation, affording the alkoxide [Ru(OCH(4-C6H4F)2)(CNN)(dppb)] (7) in 90 and 85 % yield of the isolated product. The chiral CNN-ruthenium compound [RuCl(CNN)((S,S)-Skewphos)] (8), obtained by the reaction of [RuCl2(PPh3)3] with (S,S)-Skewphos and orthometalation of HCNN in the presence of NEt3, is a highly active catalyst for the enantioselective transfer hydrogenation of methylaryl ketones (turnover frequencies (TOFs) of up to 1.4 x 10(6) h(-1) at reflux were obtained) with up to 89% ee. Also the ketone CF3CO(4-C6H4F), containing the strong electron withdrawing CF3 group, is reduced to the R alcohol with 64% ee and a TOF of 1.5 x 10(4) h(-1). The chiral alkoxide [Ru(OiPr)(CNN)((S,S)-Skewphos)]n iPrOH (9), obtained from 8 and NaOiPr in the presence of 2-propanol, reacts with CF3CO(4 C6H4F) to afford a mixture of the diastereomer alkoxides [Ru(OCH(CF3)(4 C6H4F))(CNN)((S,S)-Skewphos)] (10/11; 74% yield) with 67% de. This value is very close to the enantiomeric excess of the alcohol (R)-CF3CH(OH)(4-C6H4F) formed in catalysis, thus suggesting that diastereoisomeric alkoxides with the Ru-NH2 linkage are key species in the catalytic asymmetric transfer hydrogenation reaction. PMID- 18384029 TI - Improving sensitivity by large-volume sample stacking combined with sweeping without polarity switching by capillary electrophoresis coupled to photodiode array ultraviolet detection. AB - An easy, simple, and highly efficient on-line preconcentration method for polyphenolic compounds in CE was developed. It combined two on-line concentration techniques, large-volume sample stacking (LVSS) and sweeping. The analytes preconcentration technique was carried out by pressure injection of large-volume sample followed by the EOF as a pump pushing the bulk of low-conductivity sample matrix out of the outlet of the capillary without the electrode polarity switching technique using five polyphenols as the model analytes. Identification and quantification of the analytes were performed by photodiode array UV (PDA) detection. The optimal BGE used for separation and preconcentration was a solution composed of 10 mM borate-90 mM sodium cholate (SC)-40% v/v ethylene glycol, without pH adjustment, the applied voltage was 27.5 kV. Under optimal preconcentration conditions (sample injection 99 s at 0.5 psi), the enhancement in the detection sensitivities of the peak height and peak area of the analytes using the on-line concentration technique was in the range of 18-26- and 23-44 fold comparing with the conventional injection mode (3 s). The detection limits for (-)-epigallocatechin (EGC), (-)-epicatechin (EC), (+)-catechin (C), (-) epigallocatechin gallate (EGCG), and (-)-epicatechin gallate (ECG) were 4.3, 2.4, 2.2, 2.0, and 1.6 ng/mL, respectively. The five analytes were baseline-separated under the optimum conditions and the experimental results showed that preconcentration was well achieved. PMID- 18384030 TI - Electrokinetic supercharging for highly efficient peptide preconcentration in capillary zone electrophoresis. AB - Electrokinetic supercharging has been integrated in CZE for the development of a highly sensitive methodology for protein tryptic digest analysis. A careful choice of the experimental conditions led to sensitivity enhancement factors between 1000 and 10,000 whilst maintaining a satisfactory resolution. Peptides in the low nanomolar concentration range have been detected despite the use of the poorly sensitive UV absorbance detection mode. The buffer system used in this study is fully suitable for coupling CE to MS. PMID- 18384033 TI - Three-dimensional shape-persistent fluorescent nanocages: facile dynamic synthesis, photophysical properties, and surface morphologies. PMID- 18384034 TI - PdI2-catalyzed coupling-cyclization reactions involving two different 2,3 allenols: an efficient synthesis of 4-(1',3'-dien-2'-yl)-2,5-dihydrofuran derivatives. AB - Transition-metal-catalyzed dimeric coupling-cyclization reactions of two different 2,3-allenols afforded 4-(1',3'-dien-2'-yl)-2,5-dihydrofuran derivatives 3. 2-Substituted 2,3-allenols 1 cyclized to form the 2,5-dihydrofuran ring, whereas the 2-unsubstituted 2,3-allenols 2 provided the 1,3-diene unit at the 4 position. The reaction is proposed to proceed through an oxypalladation, insertion, and beta-hydroxide elimination process. The C=C double bond was formed with high E stereoselectivity by beta-hydroxide elimination. PMID- 18384035 TI - Structural determinants of the unusual helix stability of a de novo engineered vascular endothelial growth factor (VEGF) mimicking peptide. PMID- 18384036 TI - Tandem intramolecular hydroalkoxylation-hydroarylation reactions: synthesis of enantiopure benzofused cyclic ethers from the chiral pool. PMID- 18384037 TI - Carbonyl-substituted titanocenes: a novel class of cytostatic compounds with high antitumor and antileukemic activity. PMID- 18384038 TI - Functional quantum-dot/dendrimer nanotubes for sensitive detection of DNA hybridization. PMID- 18384039 TI - Supramolecular nanotube endo sensing for a guest protein. PMID- 18384040 TI - A multichannel electrophoresis microchip platform for rapid chiral selector screening. AB - This study presents a four-channel electrophoresis chip platform, featuring double-cross hydrostatic sample injection, for rapid chiral selector screening. This platform needs only five electrodes to drive microchip electrophoresis in four separate channels for screening four chiral selectors at a time. To demonstrate the performance of this screening platform, eight neutral CDs and their derivatives as chiral selectors were screened towards two FITC-labeled chiral compounds. The screening could be accomplished in less than 2 min. Dimethyl-beta-CD and hydroxypropyl-alpha-CD was demonstrated to be the appropriate selectors for FITC-norfenefrine and FITC-baclofen, respectively. The established platform is easy to operate and suitable for rapid screening process, which is expected to be a potential platform for high-throughput screening of chiral selectors. PMID- 18384041 TI - A high-discernment microflow cytometer with microweir structure. AB - Using a simple and reliable isotropic wet etching process, we fabricated a microflow cytometer in which cells/particles are concentrated in the center of the sample stream using a 2-D hydrodynamic focusing technique and an microweir structure. Having focused the cells/particles, they are detected and counted using a LIF method. The experimental and numerical results confirm the effectiveness of the hydrodynamic sheath flows in squeezing the cells/particles into a narrow stream in the horizontal X-Y plane. Furthermore, it is shown numerically that the microweir structure results in the separation of the cells/particles in the vertical X-Z plane such that they pass through the detection region in a sequential fashion and can therefore be counted with a high degree of precision. The experimental results obtained using fluorescent polystyrene beads with diameters of 5 and 10 microm, respectively, confirm the suitability of the proposed device for microfluidic applications requiring the high-precision counting of particles or cells within a sample flow. PMID- 18384042 TI - Microchip-based small, dense low-density lipoproteins assay for coronary heart disease risk assessment. AB - Small, dense low-density lipoprotein (sdLDL) has been accepted as an emerging cardiovascular risk factor, and there has been an increasing interest in analytical methods for sdLDL profiling for diagnosis. Serum sdLDL may be measured by different laboratory techniques, but all these methods are laborious, time consuming, and costly. Recently, we have demonstrated that a low-temperature bonding of quartz microfluidic chips for serum lipoproteins analysis (Zhuang, G., Jin, Q., Liu, J., Cong, H. et al., Biomed. Microdevices 2006, 8, 255-261). In contrast to this previous study, we chose SDS as anionic surfactant to modify both lipoproteins and the channel surface to minimize lipoprotein adsorption and improve the resolution of lipoprotein separation. Two major LDL subclass patterns including large, buoyant LDL (lLDL), sdLDL, and high-density lipoprotein (HDL) were effectively separated with high reproducibility. RSD values of the migration time (min) and peak areas of standard LDL and HDL were 6.28, 4.02, 5.02, and 2.5%, respectively. Serum lipoproteins of 15 healthy subjects and 15 patients with coronary heart disease (CHD) were separated by microchip CE. No peaks of sdLDL were detected in serum samples of healthy subjects while sdLDL fractional peaks were observed in patients' entire serum samples. These results suggested that the microchip-based sdLDLs assay was a simple, rapid, and highly efficient technique and significantly improved the analysis of CHD risk factors. PMID- 18384043 TI - Structure-based mutagenesis of Penicillium griseofulvum xylanase using computational design. AB - Penicillium griseofulvum xylanase (PgXynA) belongs to family 11 glycoside hydrolase. It exhibits unique amino acid features but its three-dimensional structure is not known. Based upon the X-ray structure of Penicillium funiculosum xylanase (PfXynC), we generated a three-dimensional model of PgXynA by homology modeling. The native structure of PgXynA displayed the overall beta-jelly roll folding common to family 11 xylanases with two large beta-pleated sheets and a single alpha-helix that form a structure resembling a partially closed right hand. Although many features of PgXynA were very similar to previously described enzymes from this family, crucial differences were observed in the loop forming the "thumb" and at the edge of the binding cleft. The robustness of the xylanase was challenged by extensive in silico-based mutagenesis analysis targeting mutations retaining stereochemical and energetical control of the protein folding. On the basis of structural alignments, modeled three-dimensional structure, in silico mutations and docking analysis, we targeted several positions for the replacement of amino acids by site-directed mutagenesis to change substrate and inhibitor specificity, alter pH profile and improve overall catalytic activity. We demonstrated the crucial role played by Ser44(PgXynA) and Ser129(PgXynA), two residues unique to PgXynA, in conferring distinct specificity to P. griseofulvum xylanase. We showed that the pH optimum of PgXynA could be shifted by -1 to +0.5 units by mutating Ser44(PgXynA) to Asp and Asn, respectively. The S44D and S44N mutants showed only slight alteration in K(m) and V(max) whereas a S44A mutant lost both pH-dependence profile and activity. We were able to produce PgXynA S129G mutants with acquired sensitivity to the Xylanase Inhibitor Protein, XIP-I. The replacement of Gln121(PgXynA), located at the start of the thumb, into an Arg residue resulted in an enzyme that possessed a higher catalytic activity. PMID- 18384044 TI - Crystal structure of archaeal tRNA(m(1)G37)methyltransferase aTrm5. AB - Methylation of the N1 atom of guanosine at position 37 in tRNA, the position 3' adjacent to the anticodon, generates the modified nucleoside m(1)G37. In archaea and eukaryotes, m(1)G37 synthesis is catalyzed by tRNA(m(1)G37)methyltransferase (archaeal or eukaryotic Trm5, a/eTrm5). Here we report the crystal structure of archaeal Trm5 (aTrm5) from Methanocaldococcus jannaschii (formerly known as Methanococcus jannaschii) in complex with the methyl donor analogue at 2.2 A resolution. The crystal structure revealed that the entire protein is composed of three structural domains, D1, D2, and D3. In the a/eTrm5 primary structures, D2 and D3 are highly conserved, while D1 is not conserved. The D3 structure is the Rossmann fold, which is the hallmark of the canonical class-I methyltransferases. The a/eTrm5-defining domain, D2, exhibits structural similarity to some class-I methyltransferases. In contrast, a DALI search with the D1 structure yielded no structural homologues. In the crystal structure, D3 contacts both D1 and D2. The residues involved in the D1:D3 interactions are not conserved, while those participating in the D2:D3 interactions are well conserved. D1 and D2 do not contact each other, and the linker between them is disordered. aTrm5 fragments corresponding to the D1 and D2-D3 regions were prepared in a soluble form. The NMR analysis of the D1 fragment revealed that D1 is well folded by itself, and it did not interact with either the D2-D3 fragment or the tRNA. The NMR analysis of the D2-D3 fragment revealed that it is well folded, independently of D1, and that it interacts with tRNA. Furthermore, the D2-D3 fragment was as active as the full length enzyme for tRNA methylation. The positive charges on the surface of D2-D3 may be involved in tRNA binding. Therefore, these findings suggest that the interaction between D1 and D3 is not persistent, and that the D2-D3 region plays the major role in tRNA methylation. PMID- 18384045 TI - Insights from the energetics of water binding at the domain-ligand interface of the Src SH2 domain. AB - SH2 domains play important roles in signal transduction by binding phosphorylated tyrosine residues on cell surface receptors. In an effort to understand the mechanism of ligand binding and more specifically the role of water, we have designed a general computational protocol based on the potential of mean force to compute the thermodynamics of water molecules at the protein-ligand interface for two SH2 domain complexes of the Src kinase, those bound to the two peptides Ac PQpYEpYI-NH2 and Ac-PQpYIpYV-NH2 where pY indicates a phosphotyrosine. These two peptides were chosen because they have similar binding affinities but very different entropic/enthalpic thermodynamic binding signatures, indicating different interactions with solvent. We find that the isoleucine to valine mutation at position +3 (the third amino acid C-terminal to pY) in the ligand has only limited impact on the water structure. By contrast, the glutamic acid to isoleucine mutation at position +1 has a significant impact by not only abrogating a local hydrophilic binding site but, more importantly and surprisingly, inducing a favorable nonlocal entropic contribution from the water molecules around the phosphorylated tyrosine at the +2 position. Our study demonstrates the validity of the method reported here for exploring the thermodynamic solvation landscape of protein-protein interactions. PMID- 18384048 TI - Investigating the role of murine epididymosomes and uterosomes in GPI-linked protein transfer to sperm using SPAM1 as a model. AB - Sperm uptake of glycosyl phosphatidylinositol (GPI)-linked proteins from luminal fluids has been shown to occur in male and estrous female reproductive tracts. In males, this is attributed to membranous vesicles secreted into the epididymis and prostate. While epididymosomes have been characterized, there have been no reports of the presence of vesicles in female luminal fluids. Here we report the presence of vesicles, characterized as "uterosomes," in the murine estrous female reproductive fluid; and use Sperm Adhesion Molecule 1 (SPAM1/PH-20), a well-known hyaluronidase found in male and female fluids, as a model to investigate vesicle mediated GPI-linked protein transfer to sperm. Epididymosomes and uterosomes isolated after ultracentrifugation of epididymal (ELF) and uterine luminal fluid (ULF) were analyzed by electron microscopy and shown to be approximately 10-70 and approximately 15-50 nm in diameter. The structural integrity of uterosomes was confirmed by their resistance to hypo-osmotic and freeze/thaw stresses; and immunogold labeling localized SPAM1 to their outer membrane surface, as was the case for epididymosomes. SPAM1 was acquired by caudal sperm during incubation in epididymosomes and uterosomes; uptake was abolished when the GPI anchor was enzymatically cleaved. Sperm analyzed by confocal and transmission electron microscopy (TEM) after incubation in fluorescently labeled vesicles revealed the label on the membrane over the acrosome and midpiece of the flagella, where SPAM1 normally resides. High magnification TEM images demonstrated vesicles juxtaposed to the sperm plasma membrane potentially transferring SPAM1. Taken together, these results implicate vesicular docking as the mechanism of vesicle-mediated GPI-linked protein transfer to sperm from murine reproductive fluids. PMID- 18384046 TI - Dementia in fragile X-associated tremor/ataxia syndrome (FXTAS): comparison with Alzheimer's disease. AB - Neurocognitive deficits in fragile X-associated tremor/ataxia syndrome (FXTAS) involve attentional control, working memory, executive functioning, and declarative and procedural learning. To date, no studies comparing FXTAS with other dementias have been done. We characterize the dementia in FXTAS, comparing it with Alzheimer's disease. Retrospective chart review of 68 adults (50 men, 18 women) with FXTAS. 20 men with FXTAS dementia were matched by age, gender, and education to patients with mild Alzheimer's dementia (AD). Neuropsychological measures were compared between the two groups: Boston Naming Test (BNT), phonemic fluency (Controlled Oral Word Association Test), digit span forward (DSF) and backward (DSB). Comparisons were based on analysis of covariance and t-tests to assess significant differences between groups. 50% of men with FXTAS and no women were cognitively impaired. On mean scores of verbal fluency (22.83 in FXTAS vs. 28.83 in AD, P = 0.112), working memory (DSB, 4.80 in AD vs. 5.41 in FXTAS, P = 0.359), and language (BNT, 48.54 in AD vs. 54.20 in FXTAS, P = 0.089), there were no significant differences. Digit span forward, measuring attention, was significantly higher in subjects with FXTAS dementia (8.59, vs. 7.10 in AD, P = 0.010). Individuals with FXTAS have significant cognitive deficits, on the order of those in AD although the cognitive profiles in these dementias are not similar. Further research is needed to outline the neuropsychiatric profile in FXTAS and the correlation of genetic markers with the progression and severity of cognitive loss. PMID- 18384049 TI - Use of combined in silico expression data and phylogenetic analysis to identify new oocyte genes encoding RNA binding proteins in the mouse. AB - During folliculogenesis, oocytes accumulate maternal mRNAs in preparation for the first steps of early embryogenesis. The processing of oocyte mRNAs is ensured by heterogeneous nuclear ribonucleoproteins (hnRNPs) genes that encode RNA binding proteins implied in mRNA biogenesis, translation, alternative splicing, nuclear exportation, and degradation. In the present work, by combining phylogenetic analyses and, when available, in silico expression data, we have identified three new oocyte-expressed genes encoding RNA binding proteins by using two strategies. Firstly, we have identified mouse orthologs of the Car1 gene, known to be involved in regulation of germ cell apoptosis in C. elegans, and of the Squid gene, required for the establishment of anteroposterior polarity in the Drosophila oocyte. Secondly, we have identified, among genes whose ESTs are highly represented in oocyte libraries, a paralog of Poly(A) binding protein- Interacting Protein 2 (Paip2) gene, known to inhibit the interaction of the Poly(A)-Binding Protein with Poly(A) tails of mRNAs. For all of these genes, the expression in oocyte was verified by in situ hybridization. Overall, this work underlines the efficiency of in silico methodologies to identify new genes involved in biological processes such as oogenesis. PMID- 18384051 TI - Twin study refining psychotic symptom dimensions as phenotypes for genetic research. AB - We investigated which psychotic symptom dimensions are likely to be most useful as phenotypes for genetic linkage and association studies. Two hundred twenty four probandwise twin pairs (106 monozygotic,118 same-sex dizygotic), where probands had psychosis, were ascertained from the Maudsley Twin Register in London. Dimensions were defined as ordinal symptom scores using the OPCRIT checklist, based on previous factor analyses of this and other samples. To qualify as a potentially useful phenotype, dimensions had to show (a) a satisfactory polychoric model fit and significant within-pair correlation in MZ pairs concordant for DSM-III-R psychosis and (b) that they could not be better defined in terms of other combinations of relevant symptoms, single symptoms, or dichotomized dimension scores. Relationships between dimension scores in twin probands and risk of psychosis in co-twins were also investigated. None of the positive or negative dimensions satisfied the phenotypic criteria. The disorganized dimensions showed significant and substantial correlations which maximized for a narrow definition. Combined negative/disorganized dimensions also showed significant and substantial correlations, but did not have advantages over disorganized dimensions. None of the dimensions were significant predictors of psychosis risk in co-twins. We conclude that, of the dimensions analyzed, the narrow disorganized dimension shows most promise as a phenotype for molecular genetic research. PMID- 18384050 TI - Lifestyle physical activity of older rural women. AB - The purpose of this study was to describe and examine the lifestyle physical activity behaviors (household, leisure, occupational) of older rural women. Background characteristics included demographics, environment, social support, and health. Intrapersonal characteristics included motivation and self-efficacy. The majority of the women's energy expenditure was in the household dimension. Social support was positively associated with household activities. A higher level of leisure physical activity was associated with living within the two small cities and reporting lower levels of health and lower motivation. This research highlights the importance of household physical activity and the contribution of social support for household physical activity, both of which may be important in developing interventions to promote physical activity in older rural women. PMID- 18384052 TI - Complete nucleotide sequence of a coxsackievirus B4 strain that establishes infection in ICR mice pancreas and induces glucose intolerance. AB - Some coxsackievirus B serotypes are potentially diabetogenic. Previous studies revealed that the virulence and the tissue damage varied with the genetics of the virus strain as well as with the genetics of the mice. A single amino acid variation can alter virulence and tropism in both murine and in vitro models. However, the genetic determinants of this phenomenon have not been determined. In this study, infections with a laboratory strain of coxsackievirus B4 resulted in a diabetes-like syndrome in ICR mice, characterized by chronic pancreatic inflammation together with dysregulation in glucose metabolism, loss of pancreatic acinar tissue and persistent infection in islets. To characterize the genetic determinants involved in the mouse pancreas adaptation, the laboratory strain of coxsackievirus B4 was cloned for molecular characterization. Comparing the whole genome sequence of this virus strain with the other coxsackievirus B4 strains revealed some differences. Altogether 15 nucleotides were changed, resulting in 10 amino acid substitutions, which might be responsible for the pathogenic phenotype of this strain in mice. PMID- 18384053 TI - Spatiotemporal expression of SSeCKS in injured rat sciatic nerve. AB - SSeCKS (src suppressed C kinase substrate) functions in the control of cell signaling and cytoskeletal arrangement. It is expressed in brain and spinal cord, but little is known about its expression in peripheral nerves. In this study, in rats, real-time polymerase chain reaction and Western blot analysis showed that expression of SSeCKS in crushed sciatic nerve reached its highest level 6 hr after crushing, whereas in a transection model, SSeCKS peaked at 2 days in the proximal stump and 12 hr in the distal stump. Immunohistochemical analysis demonstrated up-regulation of SSeCKS protein surrounding the crush site and in the two stumps of the transected nerve. In addition, SSeCKS colocalized with growth-associated protein 43 and with S100, which also changed with time after injury. These findings support the idea that SSeCKS participates in the adaptive response to peripheral nerve injury and may be associated with regeneration. PMID- 18384054 TI - Renal tumors in children: challenges for developing countries and opportunities for collaboration. PMID- 18384056 TI - The American Society of Pediatric Hematology/Oncology distinguished career award goes to Laurence A. Boxer, MD. PMID- 18384057 TI - Wilms tumour in Sudan. AB - BACKGROUND: Wilms tumour is one of the commonest childhood solid tumours which has an excellent outlook in the developed world with 5-year overall survival exceeding 90%. There is little information from Sudan regarding Wilms tumour. PROCEDURE: Records of patients with Wilms tumour diagnosed and treated at Institute of Nuclear Medicine, Molecular Biology and Oncology (INMO) in the University of Gezira from May 1999 to June 2007 were reviewed. RESULTS: Thirty seven children presented at a mean age of 4.1 years (range 2 months-13 years). The male to female ratio was 0.9-1. Abdominal swelling or mass was the commonest symptom. There was 1 child with Stage I (2.7%), 7 with stage II (18.9%), 25 with Stage III (67.6%) and 4 with Stage IV (10.8%). Following diagnosis 27% of children did not receive further treatment (5.4% died prior to treatment, 5.4% were not able to finance treatment and for the rest 16.2% no cause was identified). More than half of the children did not have a nephrectomy and only 4 (11%) completed treatment. CONCLUSIONS: The poor outlook is related to several factors. Delayed presentation, poor awareness of treatment options, lack of finances, no provision of food, lodging and transport, absence of paediatric trained staff are the obstacles to better outcomes. Empowering parents with information, giving chemotherapy prior to nephrectomy, training staff and establishing links with a tertiary cancer centre in the developed world are some of the options to improve survival. PMID- 18384058 TI - A de novo apparently balanced translocation [46,XY,t(2;9)(p13;p24)] interrupting RAB11FIP5 identifies a potential candidate gene for autism spectrum disorder. AB - Autism spectrum disorder (ASD) is a severe developmental disorder of the central nervous system characterized by impairments in social interaction, communication, and range of interests and behaviors. The syndrome's prevalence is estimated to be as high as 1 in 150 American children yet its etiology remains largely unknown. Examination of observed cytogenetic variants in individuals with ASD may identify genes involved in its pathogenesis. As part of a multidisciplinary study, an apparently balanced de novo translocation between chromosomes 2 and 9 [46,XY,t(2;9)(p13;p24)] was identified in a subject with pervasive developmental disorder not otherwise specified (PDD-NOS), and no distinctive dysmorphic features. Molecular characterization of the rearrangement revealed direct interruption of the RAB11 family interacting protein 5 (RAB11FIP5) gene. RAB11FIP5 is a Rab effector involved in protein trafficking from apical recycling endosomes to the apical plasma membrane. It is ubiquitously expressed and reported to contribute to both neurotransmitter release and neurotransmitter uptake at the synaptic junction. Detailed analysis of the rearrangement breakpoints suggests that the reciprocal translocation may have formed secondary to incorrect repair of double strand breaks (DSBs) by nonhomologous end-joining (NHEJ). PMID- 18384059 TI - Association analysis of schizophrenia on 18 genes involved in neuronal migration: MDGA1 as a new susceptibility gene. AB - Several lines of evidence support the theory of schizophrenia (SZ) being a neurodevelopmental disorder. The structural, cytoarchitectural and functional brain abnormalities reported in patients with SZ, might be due to aberrant neuronal migration, since the final position of neurons affects neuronal function, morphology, and formation of synaptic connections. We have investigated the putative association between SZ and gene variants engaged in the neuronal migration process, by performing an association study on 839 cases and 1,473 controls of Scandinavian origin. Using a gene-wide approach, tagSNPs in 18 candidate genes have been genotyped, with gene products involved in the neuron-to glial cell adhesion, interactions with the DISC1 protein and/or rearrangements of the cytoskeleton. Of the 289 markers tested, 19 markers located in genes MDGA1, RELN, ITGA3, DLX1, SPARCL1, and ASTN1, attained nominal significant P-values (P < 0.05) in either a genotypic or allelic association test. All of these genes, except transcription factor DLX1, are involved in the adhesion between neurons and radial glial cells. Eight markers obtained nominal significance in both tests, and were located in intronic or 3'UTR regions of adhesion molecule MDGA1 and previously reported SZ candidate RELN. The most significant result was attained for MDGA1 SNP rs9462341 (unadjusted association results: genotypic P = 0.00095; allelic P = 0.010). Several haplotypes within MDGA1, RELN, ITGA3, and ENAH were nominally significant. Further studies in independent samples are needed, including upcoming genome wide association study results, but our data suggest that MDGA1 is a new SZ susceptibility gene, and that altered neuronal migration is involved in SZ pathology. PMID- 18384060 TI - Expression of SV2 in the developing chick cerebellum: comparison with Calbindin and AMPA glutamate receptors 2/3. AB - The well-organized cerebellum is an ideal model to investigate the developmental appearance and localization of pre- and postsynaptic structures. One of the synaptic proteins abundant in the central nervous system and localized in presynaptic vesicle membranes is the synaptic vesicle protein 2 (SV2). SV2 was shown to be involved in priming and modulating synaptic vesicles and having an effect in epileptic diseases. So far there are no data available describing the developmental localization of this protein in the cerebellum. We followed the expression pattern of SV2 and compared it with the expression of the neuronal calcium-binding protein Calbindin and the AMPA glutamate receptor subunits 2/3 (GluR 2/3), both shown to be early expressed in the developing chick cerebellum predominantly in Purkinje cells. We detected the expression of SV2 in presynaptic terminals (mainly from climbing and mossy fibers) as soon as they are formed at embryonic day 16 in the inner molecular layer. Purkinje cells express Calbindin and GluR 2/3 in the soma and postsynaptically in the primary dendrites at this stage. With ongoing development, the pattern of SV2 expression follows the development of Purkinje cell dendrites in the molecular layer, suggesting a synaptic refinement of labeled climbing and later parallel fibers. PMID- 18384061 TI - Predicting skull loading: applying multibody dynamics analysis to a macaque skull. AB - Evaluating stress and strain fields in anatomical structures is a way to test hypotheses that relate specific features of facial and skeletal morphology to mechanical loading. Engineering techniques such as finite element analysis are now commonly used to calculate stress and strain fields, but if we are to fully accept these methods we must be confident that the applied loading regimens are reasonable. Multibody dynamics analysis (MDA) is a relatively new three dimensional computer modeling technique that can be used to apply varying muscle forces to predict joint and bite forces during static and dynamic motions. The method ensures that equilibrium of the structure is maintained at all times, even for complex statically indeterminate problems, eliminating nonphysiological constraint conditions often seen with other approaches. This study describes the novel use of MDA to investigate the influence of different muscle representations on a macaque skull model (Macaca fascicularis), where muscle groups were represented by either a single, multiple, or wrapped muscle fibers. The impact of varying muscle representation on stress fields was assessed through additional finite element simulations. The MDA models highlighted that muscle forces varied with gape and that forces within individual muscle groups also varied; for example, the anterior strands of the superficial masseter were loaded to a greater extent than the posterior strands. The direction of the muscle force was altered when temporalis muscle wrapping was modeled, and was coupled with compressive contact forces applied to the frontal, parietal and temporal bones of the cranium during biting. PMID- 18384062 TI - Differential expression of the tight junction proteins, claudin-1, claudin-4, occludin, ZO-1, and PAR3, in the ameloblasts of rat upper incisors. AB - Tight junctions (TJs) create a paracellular permeability barrier to restrict the passage of ions, small solutes, and water. Ameloblasts are enamel-forming cells that sequentially differentiate into preameloblasts, secretory, transition, and ruffle-ended and smooth-ended maturation ameloblasts (RAs and SAs). TJs are located at the proximal and distal ends of ameloblasts. TJs at the distal ends of secretory ameloblasts and RAs are well-developed zonula occludens, but other TJs are moderately developed but incomplete zonula occludens (ZO) or less-developed macula occludens. We herein examined the immunofluorescence localization of TJ proteins, 10 claudin isoforms, occludin, ZO-1, and PAR3, a cell polarity-related protein, in ameloblasts of rat upper incisors. ZO-1 and claudin-1 were detected at both ends of all ameloblasts except for the distal ends of SAs. Claudin-4 and occludin were detected at both ends of transition and maturation ameloblasts except for the distal ends of SAs. PAR3 was detected at the proximal TJs of all ameloblasts and faintly at the distal TJs of early RAs. These results indicate that functional zonula occludens formed at the distal ends of the secretory ameloblasts and RAs consisted of different TJ proteins. Therefore, the distal TJs of secretory ameloblasts and RAs may differentially regulate the paracellular permeability to create a microenvironment suitable for enamel deposition and enamel maturation, respectively. In addition, PAR3 may be principally involved in the formation and maintenance of the proximal, but not distal, TJs. PMID- 18384063 TI - Establishment of a HPLC method for preclinical pharmacokinetic study of the novel anti-Parkinson's disease candidate drug FLZ in rats. AB - An HPLC method was established and validated for the determination of compound FLZ, a synthetic novel anti-Parkinson's disease candidate drug, in rat plasma. FLZ and the internal standard bicyclol were extracted from plasma by solid-phase extraction method and analyzed on a Restek C18 column (4.6 x 250 mm, 5 microm) with a mobile phase consisting of methanol and water (60:40, v/v) at a flow rate of 1.0 mL/min. The detection wavelength was set at 320 nm. The calibration curve was linear within the concentration range from 25 to 500 ng/mL (r2 > 0.999), the limit of quantitation was 25 ng/mL and the average recovery was 92.0% with the RSD less than 5.9%. The relative standard deviation for intra- and inter-day precision was less than 3.8 and 6.9%, respectively. The established HPLC method was validated to be a simple, rapid and reliable procedure and applied to study the preclinical pharmacokinetics of FLZ in rat plasma, and it was the first time that the pharmacokinetics of FLZ had been investigated. PMID- 18384064 TI - Simultaneous determination of naringenin and hesperetin in rats after oral administration of Da-Cheng-Qi decoction by high-performance liquid chromatography tandem mass spectrometry. AB - To quantify naringenin and hesperetin in rat plasma after oral administration of Da-Cheng-Qi decoction, a famous purgative traditional Chinese medicine, a high performance liquid chromatography-tandem mass spectrometry method was developed and validated. The HPLC separation was carried out on a Zorbax SB-C(18) column using 0.1% formic acid-methanol as mobile phase and estazolam as internal standard after the sample of rat plasma had been cleaned up with one-step protein precipitation using methanol. Atmospheric pressure chemical ionization in the positive ion mode and selected reaction monitoring method was developed to determine the active components. This method was validated in terms of recovery, linearity, accuracy and precision (intra- and inter-batch variation). The recoveries of naringenin and hesperetin were 72.8-76.6 and 75.7-77.2%, respectively. Linearity in rat plasma was observed over the range of 0.5-250 ng/mL (r2 > 0.99) for both naringenin and hesperetin. The accuracy and precision were well within the acceptable range and the relative standard deviation of the measured rat plasma samples was less than 15% (n = 5). The validated method was successfully applied for the evaluation of the pharmacokinetics of naringenin and hesperetin administered to six rats. PMID- 18384065 TI - Effect of Da-Cheng-Qi decoction on the pharmacokinetics of ranitidine in rats. AB - Da Cheng Qi decoction (DCQD) is composed of Dahuang, Houpu, Zhishi and Mangxiao. It is a formula created under the theory of Chinese medicine to purge the 'evil heat' in the gastrointestitinal tract, which arises from the ileus and acute pancreatitis. The present study was conducted to evaluate the herb-drug interaction between DCQD and ranitidine, which are often co-administered in clinical practice. Ranitidine was administered orally alone or together with DCQD to rats, and plasma ranitidine concentrations were measured by HPLC. Following oral administration, ranitidine plasma levels revealed curves characterized by peaks at 1.8 and 4.2 h corresponding to ranitidine alone and ranitidine with DCQD at mean concentrations of 16.315 and 1.455 microg/mL, respectively. After ranitidine was orally dosed alone or with DCQD, the half-lives were 1.787 and 3.758 h, while the area under the concentration-time curve (0-12 h) was 28.083 and 9.826 microg/L h, respectively, suggesting that DCQD might significantly affect the pharmacokinetics of ranitidine in rats. When physicians or pharmacists administer DCQD and ranitidine, they must make a careful effort to adjust the dosage of the drug and Chinese decoction, or avoid the herb-drug co administration. PMID- 18384066 TI - Comparative analysis on microbial and rat metabolism of ginsenoside Rb1 by high performance liquid chromatography coupled with tandem mass spectrometry. AB - Ginsenoside Rb1 is an active protopanaxadiol saponin from Panax species. In order to compare the similarities and differences of microbial and mammalian metabolisms of ginsenoside Rb1, the microbial transformation by Acremonium strictum and metabolism in rats were comparatively studied. Microbial transformation of ginsenoside Rb1 by Acremonium strictum AS 3.2058 resulted in the formation of eight metabolites. Ten metabolites (M1-M10) were detected from the in vivo study in rats and eight of them were identified as the same compounds as those obtained from microbial metabolism by liquid chromatography-tandem mass spectrometry analysis and comparison with reference standards obtained from microbial metabolism. Their structures were identified as ginsenoside Rd, gypenoside XVII, 20(S)-ginsenoside Rg3, 20(R)-ginsenoside Rg3, ginsenoside F2, compound K, 12beta-hydroxydammar-3-one-20(S)-O-beta-d-glucopyranoside, and 25 hydroxyl-(E)-20(22)-ene-ginsenoside Rg3, respectively. The structures of the additional two metabolites were tentatively characterized as 20(22),24-diene ginsenoside Rg3 and 25-hydroxyginsenoside Rd by HPLC-MS/MS analysis. M7-M10 are the first four reported metabolites in vivo. The time course of rat metabolism of ginsenoside Rb1 was also investigated. PMID- 18384067 TI - Chemical cytometry on microfluidic chips. AB - Chemical cytometry, referring to the analysis of the chemical contents in individual cells, has been in intensive study since Kennedy's first work that was published in Science. The early researches relied on fine-tip capillaries to capture the cells and do the analyses, which were lab- and time-intensive and required high skills of operation. The emergence of microfluidics has greatly spurred this research field and a great number of research papers have been published in the last decades. Highly integrated microfluidic chips have been developed to capture multiple single cells, lyse them, perform chemical reactions in enclosed microchambers, separate contents by CE and detect chemical species in individual cells. This review focuses on the development of relevant components and their integration for on-chip chemical cytometry. PMID- 18384068 TI - Electrochemical techniques for microfluidic applications. AB - Electrochemical principles provide key techniques to promote the construction of bio/chemical microsystems of the next generation. There is a wealth of technology for the microfabrication of bio/chemical sensors. In addition, microfluidic transport in a network of flow channels, pH regulation, and automatic switching can be realized by electrochemical principles. Since the basic components of the devices are electrode patterns, the integration of different components is easily achieved. With these techniques, bio/chemical assays that require the exchange of solutions can be conducted on a chip. Furthermore, autonomous microanalysis systems that can carry out necessary procedures are beginning to be realized. In this article, techniques developed in our group will be comprehensively introduced. PMID- 18384069 TI - Fabrication, modification, and application of poly(methyl methacrylate) microfluidic chips. AB - Poly(methyl methacrylate) (PMMA) is particularly useful for microfluidic chips with the features of low price, excellent optic transparency, attractive mechanical and chemical properties, ease of fabrication and modification, biocompatibility, etc. During the past decade, significant progress in the PMMA microfluidic chips has occurred. This review, which contains 120 references, summarizes the recent advances and the key strategies in the fabrication, modification, and application of PMMA microfluidic chips. It is expected that PMMA microchips should find a wide range of applications and will lead to the creation of truly disposable microfluidic devices. PMID- 18384070 TI - Recent advances in surface-enhanced Raman scattering detection technology for microfluidic chips. AB - Microfluidic chip devices and their application to sensitive chemical and biological analyses have attracted significant attention over the past decade. The miniaturization of reaction systems offers practical advantages over conventional benchtop systems. In this case, however, a highly sensitive on-chip detection method is important for the monitoring of chemical reactions as well as for the detection of analytes inside the channel because the detection volume in a micrometer-size channel is extremely small. Recently, a surface-enhanced Raman scattering (SERS) technique is being regarded as a potential candidate for the highly sensitive detection of analytes in a microfluidic chip. This review provides a general survey and an in-depth look at recent developments in SERS techniques for the biological/environmental analysis of minute analytes in a microfluidic chip. PMID- 18384071 TI - Electrophoretic analysis of food dyes using a miniaturized microfluidic system. AB - A simple and sensitive on-chip preconcentration, separation, and electrochemical detection (ED) method for the electrophoretic analysis of food dyes was developed. The microchip comprised of three parallel channels: the first two are for the field-amplified sample stacking (FASS) and subsequent field-amplified sample injection (FASI) steps, while the third one is for the micellar EKC with ED (MEKC-ED) step. The food dyes were initially extracted from real samples by employing a method that was simpler, easier, and faster compared with a standard method. The extraction of the samples was characterized by UV-Vis and electrochemical experiments. The chronoamperometric detection was performed with a glassy carbon electrode coupled horizontally with the microchip at the separation channel exit. Experimental parameters affecting the analytical performance of the method were assessed and optimized. The sensitivity of the method was improved by approximately 10,800-fold when compared with a conventional MEKC-ED analysis. Reproducible response was observed during multiple injections of samples with an RSD of <7.2% (n=5). The calibration plots were linear (r2=0.998) within the range of 1.0 nM-1.0 microM for all food dyes. LODs were estimated between 1.0 and 5.0 nM, based on S/N=3, for food dyes. The applicability of the method for the analysis of food dyes in real sample was demonstrated. PMID- 18384072 TI - Functional annotation by sequence-weighted structure alignments: statistical analysis and case studies from the Protein 3000 structural genomics project in Japan. AB - A method to functionally annotate structural genomics targets, based on a novel structural alignment scoring function, is proposed. In the proposed score, position-specific scoring matrices are used to weight structurally aligned residue pairs to highlight evolutionarily conserved motifs. The functional form of the score is first optimized for discriminating domains belonging to the same Pfam family from domains belonging to different families but the same CATH or SCOP superfamily. In the optimization stage, we consider four standard weighting functions as well as our own, the "maximum substitution probability," and combinations of these functions. The optimized score achieves an area of 0.87 under the receiver-operating characteristic curve with respect to identifying Pfam families within a sequence-unique benchmark set of domain pairs. Confidence measures are then derived from the benchmark distribution of true-positive scores. The alignment method is next applied to the task of functionally annotating 230 query proteins released to the public as part of the Protein 3000 structural genomics project in Japan. Of these queries, 78 were found to align to templates with the same Pfam family as the query or had sequence identities > or = 30%. Another 49 queries were found to match more distantly related templates. Within this group, the template predicted by our method to be the closest functional relative was often not the most structurally similar. Several nontrivial cases are discussed in detail. Finally, 103 queries matched templates at the fold level, but not the family or superfamily level, and remain functionally uncharacterized. PMID- 18384073 TI - Expression of the calcitonin receptor, calcitonin receptor-like receptor, and receptor activity modifying proteins during osteoclast differentiation. AB - The expressions of the calcitonin receptor (CTR), the calcitonin receptor-like receptor (CLR), the receptor activity-modifying proteins (RAMP) 1-3, and of the receptor component protein (RCP) have been studied in mouse bone marrow macrophages (BMM) during osteoclast differentiation, induced by treatment with M CSF and RANKL. Analyses of mRNA showed that CLR and RAMP1-3, but not CTR, were expressed in M-CSF stimulated BMM. RANKL gradually increased CTR mRNA, transiently enhanced CLR and transiently decreased RAMP1 mRNA, but did not affect RAMP2, RAMP3, or RCP mRNA. However, RANKL did not affect protein levels of CLR or RAMP1-3 as assessed by Western blots or FACS analyses, whereas immunocytochemistry showed enhanced CTR protein. Analyses of cAMP production showed that BMM cells expressed functional receptors for calcitonin gene-related peptide (CGRP), amylin, adrenomedullin, and intermedin, but not for calcitonin and calcitonin receptor stimulating peptide (CRSP), but that RANKL induced the expression of receptors for calcitonin and CRSP as well. Calcitonin, CGRP, amylin, adrenomedullin, intermedin, and CRSP all down regulated the CTR mRNA, but none of the peptides caused any effects on the expression of CLR or any of the RAMPs. Our data show that BMM cells express receptors for CGRP, amylin, adrenomedullin, and intermedin and that RANKL induces the formation of receptors for calcitonin and CRSP in these cells. We also show, for the first time, that the CTR is not only down regulated by signaling through the CTR but also by the peptides signaling through CLR/RAMPs. PMID- 18384074 TI - Where shall we meet? A role for genome organisation and nuclear sub-compartments in mediating interchromosomal interactions. AB - A recent spate of examples of specific interactions between loci on separate chromosomes in mammalian nuclei has illuminated another layer of complexity in gene regulation. As the specifics of the cross-talk between interacting loci are worked out, it is also important to consider exactly how, when and where loci can ever reliably find each other within such an intricate environment. Answers may lie in how the genome is organised in relation to itself and to specialised nuclear sub-compartments. Here, we discuss how such specialised nuclear bodies may have the potential to specifically sequester loci and provide a context where interchromosomal communications can occur. PMID- 18384076 TI - Differences of the porcine amelogenin X and Y chromosome genes (AMELX and AMELY) and their application for sex determination in pigs. AB - Molecular sexing in wild and domestic animals has becoming an important issue in several fields including reproduction. X and Y chromosome-specific sequence differences of the amelogenin genes (AMELX and AMELY) have been described in different mammalian species and used for sex determination. We studied the possibility to use sequence variability between the porcine AMELX and AMELY genes for sex determination in pigs. Sequence analysis of about 400 bp of intron 3 of the porcine amelogenin genes showed the presence of a 9-10 bp deletion in AMELY gene compared to AMELX sequences. Moreover, one single nucleotide polymorphism (SNP) was detected for the AMELY sequence. Four other SNPs and 1 bp insertion differentiated three AMELX haplotypes indicating an unexpected quite high nucleotide diversity for a chromosome X region. Two sex determination assays targeting the 9-10 bp difference between AMELX and AMELY were developed. Assessment of the accuracy of the amelogenin assays to correctly sex individuals was tested on 329 pigs belonging to different breeds/lines. All analysed animals were correctly sexed with the new designed amelogenin tests. No amplification was obtained in human, cattle, goat, sheep, and horse genomic DNA. These assays can be used for sex diagnosis of small amounts of genomic DNA (20 pg) obtained from different sources including embryo biopsies, hair, meat, and other biological specimens. Thus, apart from the application in the reproduction field, these tests can be useful in several other sectors including forensics, archaeozoology, meat production, and processing as well as for quality control in sample identification. PMID- 18384075 TI - Social support in older individuals: the role of the BDNF Val66Met polymorphism. AB - Although often viewed as a purely environmental construct, perception of social support may be influenced by genetic factors. This study examined the relationship between the brain-derived neurotrophic factor (BDNF) Val66Met polymorphism and social support measures in older subjects. The sample consisted of 243 depressed and 115 nondepressed older subjects, age 60 years or older; 233 were Val66 allele homozygotes, while 125 were Met66 allele carriers. All subjects completed clinical assessments, including a self-report questionnaire assessing four social support domains, and provided blood for genotyping. Statistical models examined the relationship between scale scores of social support and BDNF Val66Met genotype, while controlling for presence or absence of major depressive disorder and other demographic factors significantly associated with social support. As social support measures were not normally distributed, log transformed scores were examined. After controlling for diagnosis and education level, the Met66 allele was associated with lower levels of subjective social support (F(1,357) = 5.33, P = 0.0216) and a trend for fewer social interactions (F(1,357) = 3.66, P = 0.0567). To our knowledge, this is the first report associating a measure of social support with a genetic polymorphism. This supports previous work that genetic factors may influence social support perception. Further work is needed to determine the generalizability of this finding to the broader population, as well as its significance for clinical outcomes. PMID- 18384077 TI - Association of TGIFLX/Y mRNA expression with azoospermia in infertile men. AB - HOX genes are well-known to encode transcriptional regulatory proteins that play essential roles in directing embryonic development. TGIFLX/Y contains two genes, TGIFLX (X-linked) and TGIFLY (Y-linked), which are specifically expressed in human adult testes. The function(s) of these genes in normal and abnormal development are unknown. To investigate the potential role(s) of the TGIFLX/Y gene in infertile males, a nested reverse transcriptase polymerase chain reaction (RT-PCR) was performed on testicular samples from 110 patients with nonobstructive azoospermia. Although the only 51 (46.4%) of the 110 patients had detectable levels of TGIFLY expression, none of the patients with various spermatogenesis defects showed any of the TGIFLX gene expression found in normal testes. These results suggest that the function of TGIFLX may be required for the regulation of spermatogonial stem cell specification and proliferation. While functional similarity has been demonstrated among some homeobox genes, these results may refute the suggestion of redundancy between TGIFLX and TGIFLY. Furthermore, TGIFLX might be a potential biomarker candidate for male infertility assessment. PMID- 18384078 TI - Association study of candidate variants of COMT with neuroticism, anxiety and depression. AB - The Val158Met polymorphism of the gene encoding catechol-O-methyltransferase (COMT) is one of the most widely tested variants for association with psychiatric disorders, but replication has been inconsistent including both sex limitation and heterogeneity of the associated allele. In this study we investigate the association between three SNPs from COMT and anxiety and depression disorders and neuroticism all measured within the same study sample. Participants were selected as sibling pairs (or multiples) that were either concordant or discordant for extreme neuroticism scores from a total sample of 18,742 Australian twin individuals and their siblings. All participants completed the Composite International Diagnostic Interview (CIDI) from which diagnoses of DSM-IV depression and anxiety disorders were determined. Of the participants, 674 had a diagnosis of anxiety and/or depression from 492 families. Study participants (n = 2,045 from 987 families) plus, where possible, their parents were genotyped for rs737865, rs4680 (Val158Met), and rs165599. Using family based tests we looked for association between these variants and neuroticism, depression, anxiety, panic disorder and agarophobia (PDAG) and obsessive compulsive disorder. We found no convincing evidence for association either in allelic or genotypic tests for the total sample or when the sample was stratified by sex. Haplotype T-G-G showed weak association (P = 0.042) with PDAG before correction for multiple testing; association between this haplotype and schizophrenia has been previously reported in an Australian sample. PMID- 18384079 TI - Combined effects of exonic polymorphisms in CRHR1 and AVPR1B genes in a case/control study for panic disorder. AB - Accumulating evidence from animal studies suggests that the corticotropin releasing hormone (CRH) and arginine vasopressin (AVP) neuropeptide systems, contribute to anxiety behavior. To investigate whether polymorphisms in the genes regulating these two systems may alter susceptibility to anxiety disorders in humans, we genotyped 71 single nucleotide polymorphisms (SNPs) in CRH, CRHR1, CRHR2, AVP, AVPR1A, AVPR1B in a German sample from Munich with patients suffering from panic disorder and matched healthy controls (n = 186/n = 299). Significant associations were then replicated in a second German sample with 173 patients with panic disorder and 495 controls. In both samples separately and the combined sample, SNPs within CHRH1 and AVPR1B were nominally associated with panic disorder. We then tested two locus multiplicative and interaction effects of polymorphisms of these two genes on panic disorder. Fifteen SNP pairs showed significant multiplicative effects in both samples. The SNP pair with the most significant association in the combined sample (P = 0.00057), which withstood correction for multiple testing, was rs878886 in CRHR1 and rs28632197 in AVPR1B. Both SNPs are of potential functional relevance as rs878886 is located in the 3' untranslated region of the CRHR1 and rs28632197 leads to an arginine to histidine amino acid exchange at position 364 of AVPR1B which is located in the intracellular C-terminal domain of the receptor. These data suggest that polymorphisms in the AVPR1B and the CRHR1 genes alter the susceptibility to panic disorder. PMID- 18384080 TI - Supercritical fluid assisted, integrated process for the synthesis and separation of different lipid derivatives. AB - The partial synthesis of ethyl esters from corn and fish oil was carried out in a stirred tank at supercritical conditions and the products were separated from the reaction mixture using the supercritical fluid. Corn oil can be almost completely converted to ethyl esters under optimized conditions and the rate of reaction is comparable to atmospheric pressure conditions. Some concentration of polyunsaturates occurs in the partially converted glyceride fraction of fish oil, as the enzyme used, Lipozyme TL-IM, allows preferential esterification of the saturates and mono-unsaturates that are concentrated in positions 1 and 3 of the triglycerides. By judicious selection of the processing conditions, good separation of ethyl esters from the partially reacted glycerides was achieved. PMID- 18384081 TI - X-ray structures of Sap1 and Sap5: structural comparison of the secreted aspartic proteinases from Candida albicans. AB - Proteolytic activity is an important virulence factor for Candida albicans (C. albicans). It is attributed to the family of the secreted aspartic proteinases (Saps) from C. albicans with a minimum of 10 members. Saps show controlled expression and regulation for the individual stages of the infection process. Distinct isoenzymes can be responsible for adherence and tissue damage of local infections, while others cause systemic diseases. Earlier, only the structures of Sap2 and Sap3 were known. In our research, we have now succeeded in solving the X ray crystal structures of the apoenzyme of Sap1 and Sap5 in complex with pepstatin A at 2.05 and 2.5 A resolution, respectively. With the structure of Sap1, we have completed the set of structures of isoenzyme subgroup Sap1-3. Of subgroup Sap4-6, the structure of the enzyme Sap5 is the first structure that has been described up to now. This facilitates comparison of structural details as well as inhibitor binding modes among the different subgroup members. Structural analysis reveals a highly conserved overall secondary structure of Sap1-3 and Sap5. However, Sap5 clearly differs from Sap1-3 by its electrostatic overall charge as well as through structural conformation of its entrance to the active site cleft. Design of inhibitors specific for Sap5 should concentrate on the S4 and S3 pockets, which significantly differ from Sap1-3 in size and electrostatic charge. Both Sap1 and Sap5 seem to play a major part in superficial Candida infections. Determination of the isoenzymes' structures can contribute to the development of new Sap-specific inhibitors for the treatment of superficial infections with a structure-based drug design program. PMID- 18384082 TI - Using a strategy based on the concept of convergent evolution to identify residue substitutions responsible for thermal adaptation. AB - Factors that are related to thermostability of proteins have been extensively studied in recent years, especially by comparing thermophiles and mesophiles. However, most of them are global characters. It is still not clear how to identify specific residues or fragments which may be more relevant to protein thermostability. Moreover, some of the differences among the thermophiles and mesophiles may be due to phylogenetic differences instead of thermal adaptation. To resolve these problems, I adopted a strategy to identify residue substitutions evolved convergently in thermophiles or mesophiles. These residues may therefore be responsible for thermal adaptation. Four classes of genomes were utilized in this study, including thermophilic archaea, mesophilic archaea, thermophilic bacteria, and mesophilic bacteria. For most clusters of orthologous groups (COGs) with sequences from all of these four classes of genomes, I can identify specific residues or fragments that may potentially be responsible for thermal adaptation. Functional or structural constraints (represented as sequence conservation) were suggested to have higher impact on thermal adaptation than secondary structure or solvent accessibility does. I further compared thermophilic archaea and mesophilic bacteria, and found that the most diverged fragments may not necessarily correspond to the thermostability-determining ones. The usual approach to compare thermophiles and mesophiles without considering phylogenetic relationships may roughly identify sequence features contributing to thermostability; however, to specifically identify residue substitutions responsible for thermal adaptation, one should take sequence evolution into consideration. PMID- 18384083 TI - The complex structure of calmodulin bound to a calcineurin peptide. AB - The activity of the protein phosphatase calcineurin (CN) is regulated by an autoinhibition mechanism wherein several domains from its catalytic A subunit, including the calmodulin binding domain (CaMBD), block access to its active site. Upon binding of Ca2+ and calmodulin (Ca2+/CaM) to CaMBD, the autoinhibitory domains dissociate from the catalytic groove, thus activating the enzyme. To date, the structure of the CN/CaM/Ca2+ complex has not been determined in its entirety. Previously, we determined the structure of a fusion protein consisting of CaM and a 25-residue peptide taken from the CaMBD, joined by a 5-glycine linker. This structure revealed a novel CaM binding motif. However, the presence of the extraneous glycine linker cast doubt on the authenticity of this structure as an accurate representation of CN/CaM binding in vivo. Thus, here, we have determined the crystal structure of CaM complexed with the 25-residue CaMBD peptide without the glycine linker at a resolution of 2.1 A. The structure is essentially identical to the fusion construction which displays CaM bound to the CaMBD peptide as a dimer with an open, elongated conformation. The N-lobe from one molecule and C-lobe from another encompass and bind the CaMBD peptide. Thus, it validates the existence of this novel CaM binding motif. Our experiments suggest that the dimeric CaM/CaMBD complex exists in solution, which is unambiguously validated using a carefully-designed CaM-sepharose pull-down experiment. We discuss structural features that produce this novel binding motif, including the role of the CaMBD peptide residues Arg-408, Val-409, and Phe-410, which work to provide rigidity to the otherwise flexible central CaM helix joining the N- and C-lobes, ultimately keeping these lobes apart and forcing "head-to-tail" dimerization to attain the requisite N- and C-lobe pairing for CaMBD binding. PMID- 18384084 TI - Analysis of the structure of human apo-S100B at low temperature indicates a unimodal conformational distribution is adopted by calcium-free S100 proteins. AB - S100B is one of the best-characterized members of the calcium-signaling S100 protein family. Most S100 proteins are dimeric, with each monomer containing two EF-hand calcium-binding sites (EF1, EF2). S100B and other S100 proteins respond to calcium increases in the cell by coordinating calcium and undergoing a conformational change that allows them to interact with a variety of cellular targets. Although several three dimensional structures of S100 proteins are available in the calcium-free (apo-) state it has been observed that these structures appear to adopt a wide range of conformations in the EF2 site with respect to the positioning of helix III, the helix that undergoes the most dramatic calcium-induced conformational change. In this work, we have determined the structure of human apo-S100B at 10 degrees C to examine whether temperature might be responsible for these structural differences. Further, we have used this data, and other available apo-S100 structures, to show that despite the range of interhelical angles adopted in the apo-S100 structures, normal Gaussian distributions about the mean angles found in the structure of human apo-S100B are observed. This finding, only obvious from the analysis of all available apo-S100 proteins, provides direct structural evidence that helix III is a loosely packed helix. This is likely a necessary functional property of the S100 proteins that facilitates the calcium-induced conformational change of helix III. In contrast, the calcium-bound structures of the S100 proteins show significantly smaller variability in the interhelical angles. This shows that calcium binding to the S100 proteins causes not only a conformational change but results in a tighter distribution of helices within the EF2 calcium binding site required for target protein interactions. PMID- 18384085 TI - Structural studies of prephenate dehydratase from Mycobacterium tuberculosis H37Rv by SAXS, ultracentrifugation, and computational analysis. AB - Tuberculosis (TB) is one of the most common infectious diseases known to man and responsible for millions of human deaths in the world. The increasing incidence of TB in developing countries, the proliferation of multidrug resistant strains, and the absence of resources for treatment have highlighted the need of developing new drugs against TB. The shikimate pathway leads to the biosynthesis of chorismate, a precursor of aromatic amino acids. This pathway is absent from mammals and shown to be essential for the survival of Mycobacterium tuberculosis, the causative agent of TB. Accordingly, enzymes of aromatic amino acid biosynthesis pathway represent promising targets for structure-based drug design. The first reaction in phenylalanine biosynthesis involves the conversion of chorismate to prephenate, catalyzed by chorismate mutase. The second reaction is catalyzed by prephenate dehydratase (PDT) and involves decarboxylation and dehydratation of prephenate to form phenylpyruvate, the precursor of phenylalanine. Here, we describe utilization of different techniques to infer the structure of M. tuberculosis PDT (MtbPDT) in solution. Small angle X-ray scattering and ultracentrifugation analysis showed that the protein oligomeric state is a tetramer and MtbPDT is a flat disk protein. Bioinformatics tools were used to infer the structure of MtbPDT. A molecular model for MtbPDT is presented and molecular dynamics simulations indicate that MtbPDT is stable. Experimental and molecular modeling results were in agreement and provide evidence for a tetrameric state of MtbPDT in solution. PMID- 18384086 TI - Towards an integrated description of hydrogen bonding and dehydration: decreasing false positives in virtual screening with the HYDE scoring function. AB - We developed a new empirical scoring function, HYDE, for the evaluation of protein-ligand complexes. HYDE estimates binding free energy based on two terms for dehydration and hydrogen bonding only. The essential feature of this scoring function is the integrated use of log P-derived atomic increments for the prediction of free dehydration energy and hydrogen bonding energy. Taking the dehydration of atoms within the interface into account shows that some atoms contribute favorably to the overall score, while others contribute unfavorably. For instance, hydrogen bond functions are penalized if they are dehydrated unless they can overcompensate this loss by forming a hydrogen bond with excellent geometry. The main stabilizing contribution represents the removal of apolar groups from the water: the hydrophobic effect. Initial studies using the DUD dataset show that with HYDE, there is a significant decrease in false positives, a reasonable categorization of compounds as either non-binders, weak, medium or strong binders, and in particular, there is a generally applicable and thermodynamically sensible cutoff score below which there is a high likelihood that the compound is indeed a binder. PMID- 18384087 TI - Is there any place for alimentary probiotics, prebiotics or synbiotics, for patients with inflammatory bowel disease? AB - The pathogenesis of inflammatory bowel disease (IBD) involves an interaction between genetically determined host susceptibility, dysregulated immune response, and the enteric microbiota. Ecological treatments including probiotics, prebiotics, and synbiotics are actively studied in Crohn's disease (CD), ulcerative colitis (UC) and pouchitis. We review herein the literature on the rational use of probiotics in IBD considering efficacy (as evaluated in randomized controlled trials), mechanisms of action and safety issues. A probiotic effect is strictly restricted to one defined strain and cannot be generalized from one to another. There is evidence of efficacy of some probiotic drugs in pouchitis (VSL#3), and in the prevention of recurrence of UC (Escherichia coli Nissle 1917). However, the evidence for efficacy of probiotic drugs in CD is still low as well as that of dietary ecological treatments. Despite an ecological (hopefully nutritional) treatment of IBD is promising, many questions remain unanswered and further clinical and fundamental studies are needed. PMID- 18384088 TI - 6-Shogaol induces apoptosis in human colorectal carcinoma cells via ROS production, caspase activation, and GADD 153 expression. AB - Ginger, the rhizome of Zingiber officinale, is a traditional medicine with anti inflammatory and anticarcinogenic properties. This study examined the growth inhibitory effects of the structurally related compounds 6-gingerol and 6-shogaol on human cancer cells. 6-Shogaol [1-(4-hydroxy-3-methoxyphenyl)-4-decen-3-one] inhibits the growth of human cancer cells and induces apoptosis in COLO 205 cells through modulation of mitochondrial functions regulated by reactive oxygen species (ROS). ROS generation occurs in the early stages of 6-shogaol-induced apoptosis, preceding cytochrome c release, caspase activation, and DNA fragmentation. Up-regulation of Bax, Fas, and FasL, as well as down-regulation of Bcl-2 and Bcl-X(L )were observed in 6-shogaol-treated COLO 205 cells. N acetylcysteine (NAC), but not by other antioxidants, suppress 6-shogaol-induced apoptosis. The growth arrest and DNA damage (GADD)-inducible transcription factor 153 (GADD153) mRNA and protein is markedly induced in a time- and concentration dependent manner in response to 6-shogaol. PMID- 18384089 TI - Inhibitory effects of trans-resveratrol analogs molecules on the proliferation and the cell cycle progression of human colon tumoral cells. AB - Resveratrol may function as a cancer chemopreventive agent. However, few data are available on the antitumoral activities of its dimer, epsilon-viniferin, also present in human diet. So, the effects of resveratrol, epsilon-viniferin, of their acetylated forms (resveratrol triacetate, epsilon-viniferin pentaacetate) and of vineatrol (a wine grape extract) were compared on human adenocarcinoma colon cells. Resveratrol and resveratrol triacetate inhibit cell proliferation and arrest cell cycle. epsilon-Viniferin and epsilon-viniferin pentaacetate slightly reduce cell proliferation. Vineatrol inhibits cell proliferation and favors an accumulation in the S phase of the cell cycle. Consequently, resveratrol triacetate and vineatrol could constitute new putative anticancer agents on colon carcinoma. PMID- 18384090 TI - Grape seed proanthocyanidines and skin cancer prevention: inhibition of oxidative stress and protection of immune system. AB - Overexposure of the skin to UV radiation has a variety of adverse effects on human health, including the development of skin cancers. There is a need to develop nutrition-based efficient chemopreventive strategies. The proanthocyanidins present in grape seeds (Vitis vinifera) have been shown to have some biological effects, including prevention of photocarcinogenesis. The present communication discusses the in vitro and in vivo studies of the possible protective effect of grape seed proanthocyanidins (GSPs) and the molecular mechanism for these effects. In SKH-1 hairless mice, dietary supplementation with GSPs is associated with a decrease of UVB-induced skin tumor development in terms of tumor incidence, tumor multiplicity, and a decrease in the malignant transformation of papillomas to carcinomas. It is suggested that the chemopreventive effects of dietary GSPs are mediated through the attenuation of UV-induced: (i) oxidative stress; (ii) activation of mitogen-activated protein kinases and nuclear factor-kappa B (NF-kappaB) signaling pathways; and (iii) immunosuppression through alterations in immunoregulatory cytokines. Collectively, these studies indicate protective potential of GSPs against experimental photocarcinogenesis in SKH-1 hairless mice, and the possible mechanisms of action of GSPs, and suggest that dietary GSPs could be useful in the attenuation of the adverse UV-induced health effects in human skin. PMID- 18384091 TI - Effect of artichoke extract (Cynara scolymus L.) on palmitic-1-14C acid oxidation in rats. AB - Studies on the effect of the artichoke extract (AE) on oxidation of palmitic-1 14C acid administered intravenously to rats at a dose 25 and 50 mg/kg bw demonstrated marked enhancement of both 14CO2 expiration rate and 14CO2 recovery in the expired air. The extract suppressed accumulation of palmitic-1-14C acid in serum lipids and epididymal fat pad tissue as well. The effects of the extract on 14CO2 expiration rate, 14CO2 recovery, as well as accumulation of palmitic-1-14C acid were dose dependent. Total14CO2 recovery in expired air during 60 min was elevated by 17.3% (p < 0.05) and 52.1% (p < 0.001) in rats administered the extract at a dose of 25 and 50 mg/kg, respectively. The rats supplemented with the AE at a dose of 25 and 50 mg/kg bw were characterized by 10.0% (not significant) and 19% (p < 0.05) decrease in( 14)C radioactivity of serum lipids as well as reduction of epididymal fat tissue 14C radioactivity by 8.7 and 17.5% (p < 0.05), respectively, in comparison with the control rats. Thus, the results demonstrate that the AE possess stimulatory properties with respect to oxidation of palmitic acid administered to rats, and provide new information on the mechanism of antilipemic activity of the extract associated with activation of lipid oxidation in the organism. PMID- 18384092 TI - Production and characterization of an allergen panel for component-resolved diagnosis of celery allergy. AB - In celery a relevant food allergen source, three allergens have been identified so far: Api g 1 and Api g 4, and one glycosylated protein, Api g 5. For component resolved food allergy diagnosis high amounts of well-defined allergens are needed. Depending on the individual celery allergen, protocols for heterologous production and purification from natural source, respectively, were established to obtain homogenous protein batches. Afterwards the purified recombinant allergens, Api g 1, Api g 4 and natural Api g 5 were characterized regarding their structural integrity and immunological activity. Therefore, several methods were applied. Proteins were identified by partial N-terminal sequencing, protein mass was verified by MS and sequence integrity by MALDI-TOF and N-terminal sequencing after tryptic digestion. Presence of isoforms in natural allergen preparations was identified by 2-DE. Secondary and tertiary structures were evaluated by circular dichroism (CD) spectroscopy and NMR analysis. Finally, IgE binding capacity was verified using selected sera from celery allergic patients in IgE immunoblots and IgE ELISA. These well-defined celery allergens will be used to prove the concept of component-resolved diagnosis and will contribute to improve food allergy diagnosis in the future. PMID- 18384093 TI - Purification and structural stability of the peach allergens Pru p 1 and Pru p 3. AB - Pru p 1 (a Bet v 1 homologue) and Pru p 3 (a nonspecific lipid transfer protein; nsLTP) are major allergenic proteins in peach fruit, but differ in their abundance and stability. Pru p 1 has low abundance and is highly labile and was purified after expression as a recombinant protein in Escherichia coli. Pru p 3 is highly abundant in peach peel and was purified by conventional methods. The identities of the proteins were confirmed by sequence analysis and their masses determined by MS analysis. The purified proteins reacted with antisera against related allergens from other species: Pru p 1 with antiserum to Bet v 1 and Pru p 3 with antiserum to Mal d 3 (from apple). The presence of secondary and tertiary structure was demonstrated by circular dichroism (CD) and high field NMR spectroscopy. CD spectroscopy also showed that the two proteins differed in their stability at pH 3 and in their ability to refold after heating to 95 degrees C. Thus, Pru p 1 was unfolded at pH 3 even at 25 degrees C but was able to refold after heating to 95 degrees C at pH 7.5. In contrast, Pru p 3 was unable to refold after heating under neutral conditions but readily refolded after heating at pH 3. PMID- 18384094 TI - Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA. AB - Thaumatin-like proteins (TLPs) have been established as a new family of fruit and pollen allergens. The aim of this study was to develop a two-site ELISA for the quantification of the thaumatin-like kiwi allergen (Act d 2) in kiwifruit extracts and kiwifruit-containing food products. Genomic DNA (gDNA) of Act d 2 was amplified and the deduced amino acid sequence was determined to obtain a primary structure. Act d 2 purified from kiwifruit extract by HPLC was identified by Edman degradation and MS. Balb/c mice were immunized with Act d 2 for the production of mAbs by hybridoma technology. The optimized ELISA measured Act d 2 concentrations ranging from 0.2 to 9.0 ng/mL, with intra- and interassay coefficients of variation of 3.65 and 10.44%, respectively. The developed ELISA is a useful method for the quantification of the thaumatin-like kiwi allergen in kiwifruit extracts as well as the allergen level in kiwifruit-containing food products. It may be a helpful analytical tool for the evaluation of the stability (integrity) of fruit allergen extracts for in vitro diagnosis. PMID- 18384095 TI - Erythrodiol, a natural triterpenoid from olives, has antiproliferative and apoptotic activity in HT-29 human adenocarcinoma cells. AB - Erythrodiol is the precursor of pentacyclic triterpenic acids present in Olea Europaea. Although olive oil and some of its constituents are reported to have anticarcinogenic activities, erythrodiol has not been assessed in its cell biological functions in detail. We therefore determined its effects on cell growth and apoptosis in human colorectal carcinoma HT-29 cells. Proliferation, cytotoxicity, and apoptosis were measured by fluorescence-based techniques. Erythrodiol inhibited cell growth with an EC50 value of 48.8 +/- 3.7 microM without any cytotoxic effects in a concentration range up to 100 microM. However, exposure of cells for 24 h to 50, 100, and 150 microM erythrodiol increased caspase-3-like activity by 3.2-, 4.8-, and 5.2-fold over that in control cells. We here demonstrate for the first time that, in colon adenocarcinoma cells, erythrodiol exerts antiproliferative and proapoptotic activity. PMID- 18384096 TI - S100B Protein concentration in milk-formulas for preterm and term infants. Correlation with industrial preparation procedures. AB - Human milk S100B protein possesses important neurotrophic properties. However, in some conditions human milk is substituted by milk formulas. The aims of the present study were: to assess S100B concentrations in milk formulas, to verify any differences in S100B levels between preterm and term infant formulas and to evaluate the impact of industrial preparation at predetermined phases on S100B content. Two different set of samples were tested: (i) commercial preterm (n = 36) and term (n = 36) infant milk formulas; ii) milk preterm (n = 10) and term infant (n = 10) formulas sampled at the following predetermined industrial preparation time points: skimmed cow milk (Time 0); after protein sources supplementation (Time 1); after pasteurization (Time 2); after spray-drying (Time 3). Our results showed that S100B concentration in preterm formulas were higher than in term ones (p < 0.01). In addition, S100B concentrations during industrial preparation showed a significant increase (p < 0.001) at Time 1 followed by a slight decrease (p > 0.05) at Time 2, whereas a significant (p < 0.001) dip was observed at Time 3. In conclusion, S100B showed a sufficient thermostability to resist pasteurization but not spry-drying. New feeding strategies in preterm and term infants are therefore warranted in order to preserve S100B protein during industrial preparation. PMID- 18384098 TI - Multi-targeted therapy by curcumin: how spicy is it? AB - Although traditional medicines have been used for thousands of years, for most such medicines neither the active component nor their molecular targets have been very well identified. Curcumin, a yellow component of turmeric or curry powder, however, is an exception. Although inhibitors of cyclooxygenase-2, HER2, tumor necrosis factor, EGFR, Bcr-abl, proteosome, and vascular endothelial cell growth factor have been approved for human use by the United States Food and Drug Administration (FDA), curcumin as a single agent can down-regulate all these targets. Curcumin can also activate apoptosis, down-regulate cell survival gene products, and up-regulate p53, p21, and p27. Although curcumin is poorly absorbed after ingestion, multiple studies have suggested that even low levels of physiologically achievable concentrations of curcumin may be sufficient for its chemopreventive and chemotherapeutic activity. Thus, curcumin regulates multiple targets (multitargeted therapy), which is needed for treatment of most diseases, and it is inexpensive and has been found to be safe in human clinical trials. The present article reviews the key molecular mechanisms of curcumin action and compares this to some of the single-targeted therapies currently available for human cancer. PMID- 18384097 TI - The influence of selenium on immune responses. AB - Selenium (Se) is a potent nutritional antioxidant that carries out biological effects through its incorporation into selenoproteins. Given the crucial roles that selenoproteins play in regulating reactive oxygen species (ROS) and redox status in nearly all tissues, it is not surprising that dietary Se strongly influences inflammation and immune responses. The notion that Se "boosts" the immune system has been supported by studies involving aging immunity or protection against certain pathogens. However, studies examining the effects of Se status on other types of immunity such as antiparasitic responses or allergic asthma have suggested more Se may not always be beneficial. In this review, we summarize and compare the available data regarding how the levels of Se affect different types of immunity. Overall, determining how Se intake differentially affects various types of immune responses and dissecting the mechanisms by which this occurs will lead to a better utilization of Se-supplementation for human diseases involving the immune system. PMID- 18384099 TI - Application of supercritical fluid chromatography to the analysis of hydrophobic metabolites. AB - This review describes the usefulness of supercritical fluid chromatography (SFC) in the analysis of hydrophobic metabolites. The use of SFC for the analysis of naturally occurring polyprenols markedly improves the chromatographic resolution of polyprenol homologues and their geometric isomers as compared to conventional HPLC. Under optimized SFC conditions, individual homologues with 10-100-mers were separated. Furthermore, we established an analytical system for the fingerprinting and profiling of diverse lipids through the usage of SFC-MS. When a cyanopropylated silica gel packed column was used for the separation, 14 lipids were successfully detected, and the time required for analysis was less than 15 min. The use of an octadecylsilylated column for the separation depended on the differences in the fatty acid side chains. SFC is a useful separation technology for hydrophobic metabolites, which are difficult to be separated by HPLC. PMID- 18384100 TI - Pharmaceutical analysis by supercritical fluid chromatography: Optimization of the mobile phase composition on a 2-ethylpyridine column. AB - The separation of neutral, acidic, and basic pharmaceuticals with diverse physicochemical properties by packed column supercritical fluid chromatography (pSFC) on a 2-ethylpyridine column (25 cmx4.6 mm id, 3 mum particles) is presented. The optimization strategy involved separations at 100% methanol (MeOH) and at 50% MeOH/50% ACN while keeping the peak symmetry additives formic acid (FA) and isopropylamine (IPA) at constant levels of 0.25% v/v. By plotting the adjusted retention times as a function of the MeOH/ACN ratio, an optimal modifier ratio composition of 65% MeOH/35% ACN was found. The total set of 26 neutral, acidic, and basic pharmaceuticals was analyzed and the optimal composition experimentally verified. This mobile phase composition is currently used in pharmaceutical method development and open-access generic screening environments. PMID- 18384101 TI - The proteome of gastric lymph in normal and nematode infected sheep. AB - Lymph node cannulation allows the collection of lymph draining from a defined anatomical region. Proteomic analysis of that lymph offers a potentially valuable insight into the immunoinflammatory response of that particular region. In this study, ovine gastric lymph has been used to monitor the proteomic changes occurring in the tissue fluid of the abomasum, in response to infection with the parasitic nematode, Teladorsagia circumcincta. Lymph, collected temporally over an experimental infection period, was analysed by means of 2-DE and subsequent gel analysis using densitometry software. In addition, the composition of the lymphatic proteome was further explored by means of MALDI-TOF and MS/MS analyses. The concentration of gelsolin, alpha-1 beta glycoprotein and haemopexin were altered significantly (p<0.05) with infection. PMID- 18384102 TI - Proteomic analysis of the venom from the fish eating coral snake Micrurus surinamensis: novel toxins, their function and phylogeny. AB - The protein composition of the soluble venom from the South American fish-eating coral snake Micrurus surinamensis surinamensis, here abbreviated M. surinamensis, was separated by RP-HPLC and 2-DE, and their components were analyzed by automatic Edman degradation, MALDI-TOF and ESI-MS/MS. Approximately 100 different molecules were identified. Sixty-two components possess molecular masses between 6 and 8 kDa, are basically charged molecules, among which are cytotoxins and neurotoxins lethal to fish (Brachidanios rerio). Six new toxins (abbreviated Ms1 Ms5 and Ms11) were fully sequenced. Amino acid sequences similar to the enzymes phospholipase A2 and amino acid oxidase were identified. Over 20 additional peptides were identified by sequencing minor components of the HPLC separation and from 2-DE gels. A functional assessment of the physiological activity of the six toxins was also performed by patch clamp using muscular nicotinic acetylcholine receptor assays. Variable degrees of blockade were observed, most of them reversible. The structural and functional data obtained were used for phylogenetic analysis, providing information on some evolutionary aspects of the venom components of this snake. This contribution increases by a factor of two the total number of alpha-neurotoxins sequenced from the Micrurus genus in currently available literature. PMID- 18384103 TI - Predicting left ventricular remodeling after a first myocardial infarction by plasma proteome analysis. AB - Recent improvements in therapeutic strategies did not prevent left ventricular remodeling (LVR), which remains a common event (30%) after acute myocardial infarction (AMI). We report the use of a systematic approach, based on comparative proteomics, to select circulating biomarkers that may be associated with LVR. We selected 93 patients enrolled in a prospective study. These patients with anterior wall Q-wave AMI underwent echocardiographic follow-up at hospitalization, 3 months and 1 year after AMI. They were divided into three groups (no, low, or high remodeling). Plasma samples of these patients (day 5 of hospitalization) were processed and stored at -80 degrees C within 2 h and analyzed using SELDI-TOF protein chip technology. This systematic approach allowed to select candidate proteins modulated by LVR: post-translational variants of alpha1-chain of haptoglobin (Hpalpha1) corresponding to m/z 9493, 9565, and 9623, which were more elevated in remodeling patients. The peak 9493 m/z was shown having a receiving-operating characteristic (ROC) value of 0.71 between non- and remodeling patients. SELDI-TOF approach may lead to the identification of circulating proteins associated with LVR. Whether these candidate proteins will help to identify patients who are at high risk of heart failure after AMI will have to be tested in future studies. PMID- 18384104 TI - Enhanced nuclear proteomics. AB - Nuclear proteomics provides an opportunity to examine protein effectors that contribute to cellular phenotype. Both the quality and sensitivity of gel-based nuclear proteomics are limited, however, by the over-representation of histones in the protein mixture. These highly charged proteins overshadow rare species and interfere with IEF. A nuclear isolation and protein extraction procedure, tested on human embryonic stem cells, is reported that effectively isolates intact nuclei and then depletes the sample of histones by taking advantage of their ability to form an insoluble complex with DNA at lower pH (even under denaturing conditions). Ubiquitous histones and abundant nuclear actin, are depleted up to 99 +/- 0.02 and 42 +/- 5%, respectively. This technique greatly improves electrofocusing efficacy and nearly doubles the number of detected protein spots. This approach to nuclear protein isolation for 2-D PAGE opens the door to better investigation of nuclear protein dynamics. PMID- 18384105 TI - Investigating copper-regulated protein expression in Menkes fibroblasts using antibody microarrays. AB - Neurodegenerative illnesses are characterized by aberrant metabolism of biometals such as copper (Cu), zinc (Zn) and iron (Fe). However, little is known about the metabolic effects associated with altered metal homeostasis. In this study, we used an in vitro model of altered Cu homeostasis to investigate how Cu regulates cellular protein expression. Human fibroblasts containing a natural deletion mutation of the Menkes (MNK) ATP7A Cu transporter (MNK deleted) were compared to fibroblasts overexpressing ATP7A (MNK transfected). Cultures of MNK-transfected (Low-Cu) cells exhibited 95% less intracellular Cu than MNK-deleted (High-Cu) cells. Comparative proteomic analysis of the two cell-lines was performed using antibody microarrays, and significant differential protein expression was observed between Low-Cu and High-Cu cell-lines. Western blot analysis confirmed the altered protein expression of Ku80, nexilin, L-caldesmon, MAP4, Inhibitor 2 and DNA topoisomerase I. The top 50 altered proteins were analysed using the software program Pathway Studio (Ariadne Genomics) and revealed a significant over-representation of proteins involved in DNA repair and maintenance. Further analysis confirmed that expression of the DNA repair protein Ku80 was dependent on cellular Cu homeostasis and that Low-Cu levels in fibroblasts resulted in elevated susceptibility to DNA oxidation. PMID- 18384106 TI - A simple and inexpensive on-column frit fabrication method for fused-silica capillaries for increased capacity and versatility in LC-MS/MS applications. AB - A modified sol-gel method for a one-step on-column frit preparation for fused silica capillaries and its utility for peptide separation in LC-MS/MS is described. This method is inexpensive, reproducible, and does not require specialized equipments. Because the frit fabrication process does not damage polyimide coating, the frit-fabricated column can be tightly connected on-line for high pressure LC. These columns can replace any capillary liquid transfer tubing without any specialized connections up-stream of a spray tip column. Therefore multiple columns with different phases can be connected in series for one- or multiple-dimensional chromatography. PMID- 18384107 TI - Applications in brain proteomics: 8(th) HUPO Brain Proteome Project Workshop 7 October 2007, Seoul, Korea. AB - What are the current approaches in brain proteomics? Can we combine different, but complementary study designs to obtain better results concerning brain diseases? What are the neuro-hotspots, especially in Korea? These were some of the questions the participants of the 8(th) HUPO Brain Proteome Project Workshop tried to answer prior to the 6(th) HUPO World Congress in Seoul, Korea. Around 100 scientists came together during the afternoon of 7 October, 2007, to discuss and to catch up on the latest results and strategies concerning Huntington's disease, glioblastoma and standardization. PMID- 18384108 TI - The 6(th) East Midlands Proteomics Workshop supported by the BSPR and BMSS 14 November 2007, Nottingham, UK. AB - Now in its 6(th) year, the East Midlands Proteomics workshop held in November 2007 brought together over 200 scientists with a common interest in proteomic techniques and their application to complex biological and biomedical problems. For the first time, this meeting was jointly supported by the British Society for Proteome Research (BSPR) and British Mass Spectrometry Society (BMSS). PMID- 18384109 TI - Benfotiamine exhibits direct antioxidative capacity and prevents induction of DNA damage in vitro. AB - BACKGROUND: Complications in diabetes mellitus are partially mediated by enhanced formation of reactive oxygen species. Among the factors involved in reactive oxygen species formation, advanced glycation end products play a key role. Owing to a reduced activity of the enzyme transketolase, which requires diphosphorylated thiamine (vitamin B(1)) as cofactor, an accumulation of those deleterious glucose metabolites especially in diabetic patients can be observed. Benfotiamine, a lipophilic thiamine diphosphate prodrug, prevented early renal and retinal changes in animal studies, and reduced neuropathic pain in clinical studies. Several mechanisms for these activities have been described. We investigated for the first time direct antioxidant abilities of benfotiamine. Additionally, a potential DNA protective effect of benfotiamine was analysed. METHODS: Oxidative stress was detected by flow cytometry, antioxidative capacity was measured with the ferric reducing ability of plasma (FRAP) assay, two endpoints for genomic damage were assessed: the comet assay and the micronucleus test, and the expression and activity of transketolase was quantified. RESULTS: Benfotiamine prevented oxidative stress induced by the mutagen 4-nitroquinoline-1 oxide (NQO), the uremic toxin indoxyl sulfate, and the peptide hormone angiotensin II in three different kidney cell lines. Cell-free experiments showed a direct antioxidant effect of benfotiamine, which might account for the protective effect. Oxidative DNA damage, induced by angiotensin II, was completely prevented by benfotiamine. Incubation with benfotiamine increased transketolase expression and activity in the cells. CONCLUSIONS: Benfotiamine shows a direct antioxidant action. This effect of benfotiamine may be involved in the improvement of diabetic late complications, including peripheral neuropathy. PMID- 18384110 TI - Open bypass and endoluminal therapy: complementary techniques for revascularization in diabetic patients with critical limb ischaemia. AB - The use of endovascular therapy (EVT) for lower extremity atherosclerosis is markedly increasing while open surgical bypass is in decline. The results of EVT for critical limb ischaemia (CLI) are difficult to evaluate, especially for patients with diabetes. To date, only one randomized, prospective trial has been published comparing EVT with open bypass for CLI. Although early costs and outcomes were equivalent or superior for EVT, after 2 years, surgery was associated with a significantly reduced risk of future amputation and death.Approximately, 40-50% of diabetic patients with CLI can be initially treated with EVT. Patients with Trans-Atlantic Inter-Society Consensus (TASC) A and B lesions should be treated endoluminally. EVT should be used with caution in patients with TASC C and D lesions; however, in selected patients, particularly if vein conduit is lacking and life expectancy is short, EVT is not unreasonable. For low-to-moderate risk patients with TASC C or D lesions, extensive tibial disease, and suitable vein conduit, surgical bypass remains the best limb preservation option. The primary therapeutic goals are relief of rest pain, healing of ischaemic lesions, and maintenance of functional status. Haemodynamic assessment is critical following both open and EVT for CLI and aids in determining the need for further revascularization; additional interventions are required in 20-30% of CLI patients depending on the degree of ischaemia, anatomical disease extent, and mode of initial therapy. At the University of Arizona, we currently recommend that TASC A and B CLI patients undergo EVT first. TASC C and D patients should undergo bypass unless available conduit is poor, surgical risk is prohibitive, or life expectancy is limited. CLI is a serious end of-life condition given the sobering realization that only 50-55% of CLI patients are alive with an intact limb 5 years after initial presentation. PMID- 18384111 TI - The foot in perspective. AB - The diabetic foot constitutes a tremendous challenge for patients, caregivers and the health care system. The International Consensus Document of 1999 was a milestone in the recognition of the importance and consequences of the diabetic foot. Since then, many original papers have been published in this area. Large cohort studies have given us a deeper understanding regarding factors related to the outcome of diabetic foot ulcers: according to these studies, the severity of diabetic foot ulcers is greater than previously reported. More than 50% of individuals' foot ulcers have signs of infection at admission, and one-third have signs of both peripheral artery disease (PAD) and infection. The co-morbidities increase significantly with increasing severity of the foot disease. However, the trend in all these studies is a successive improvement in healing rate (50-60% at 20 weeks follow-up, > 75% at 1 year). It is important to differentiate between neuropathic and neuro-ischaemic ulcers with regard to factors related to outcome and co-morbidities.Recent research has emphasized the importance of psychological factors in the development and outcome of diabetic foot ulcers. Studies have shown that perceptions of the individual's own risks based on symptoms, and their own beliefs in the efficacy of self-care, can affect foot-care practice.The importance and influence of the health care organization and reimbursement should not be underestimated, both in the prevention and management of diabetic foot lesions. The diabetic foot should be considered a lifelong condition, as having had one ulcer dramatically increases the risk of developing a new ulcer. In an individual with diabetes and a foot ulcer, the ulcer should be considered as a sign of multi-organ disease, and a holistic approach to both management and prevention is recommended. PMID- 18384112 TI - Hypoxia stimulates the autocrine regulation of migration of vascular smooth muscle cells via HIF-1alpha-dependent expression of thrombospondin-1. AB - The migration of vascular smooth muscle cells from the media to intima and their subsequent proliferation are critical causes of arterial wall thickening. In atherosclerotic lesions increases in the thickness of the vascular wall and the impairment of oxygen diffusion capacity result in the development of hypoxic lesions. We investigated the effect of hypoxia on the migration of human coronary artery smooth muscle cells (CASMCs) via HIF-1alpha-dependent expression of thrombospondin-1 (TSP-1). When the cells were cultured under hypoxic conditions, mRNA and protein levels of TSP-1, and mRNA levels of integrin beta(3) were increased with the increase in HIF-1alpha protein. DNA synthesis and migration of the cells were stimulated under the conditions, and a neutralizing anti-TSP-1 antibody apparently suppressed the migration, but not DNA synthesis. The migration was also inhibited by RGD peptide that binds to integrin beta(3). Furthermore, the migration was completely suppressed in HIF-1alpha-knockdown cells exposed to hypoxia, while it was significantly enhanced in HIF-1alpha overexpressing cells. These results suggest that the hypoxia induces the migration of CASMCs, and that the migration is elicited by TSP-1 of which induction is fully dependent on the stabilization of HIF-1alpha, in autocrine regulation. Thus we suggest that HIF-1alpha plays an important role in the pathogenesis of atherosclerosis. PMID- 18384114 TI - Temperament, character, and dissociation among detoxified male inpatients with alcohol dependency. AB - The aim of this study was to determine possible relationships of pathological dissociation with temperament, character, and concurrent psychopathological features in a consecutive series of male alcohol-dependent patients. Fifty-eight patients with pathological dissociation were compared with 118 nondissociative patients classified by dissociative taxon membership. Beside higher scores on anxiety, depression, and alcoholism scales, a larger proportion of dissociative group reported childhood abuse, suicide attempts, and self-mutilation than did the nondissociative group. They also had higher scores of novelty seeking and harm avoidance, but lower scores of persistence, self-directedness, and cooperativeness. Trait anxiety, depression, and severity of alcoholism predicted dissociative experiences; however, none of the temperament or character measures did. Rather than being a derivative of temperament or character features, dissociative experiences of male alcohol-dependent patients are associated with overall concurrent psychopathology. PMID- 18384113 TI - 2-methoxyestradiol-induced cell death in osteosarcoma cells is preceded by cell cycle arrest. AB - 2-Methoxyestradiol (2-ME), a naturally occurring mammalian metabolite of 17beta Estradiol (E2), induces cell death in osteosarcoma cells. To further understand the molecular mechanisms of action, we have investigated cell cycle progression in 2-ME-treated human osteosarcoma (MG63, SaOS-2 and LM7 [corrected]) cells. At 5 microM, 2-ME induced growth arrest by inducing a block in cell cycle; 2-ME treatment resulted in 2-fold increases in G1 phase cells and a decrease in S phase cells in MG63 and SaOS-2 osteosarcoma cell lines, compared to the appropriate vehicle controls. 2-ME-treatment induced a threefold increase in the G2 phase in LM7 [corrected] osteosarcoma cells. The results demonstrated steroid specificity, as the tumorigenic metabolite, 16alpha-hydroxyestradiol (16-OHE), did not have any effect on cell cycle progression in osteosarcoma cells. The cell cycle arrest coincided with an increase in expression of the cell cycle markers p21, p27 and p53 proteins in 2-ME-treated osteosarcoma cells. Also, MG63 cells, transiently transfected with cDNA for a 'loss of function mutant' RNA-dependent protein kinase (PKR) protein, were resistant to 2-ME-induced cell cycle arrest. These results suggest that 2-ME works in concert with factors regulating cell cycle progression, and cell cycle arrest precedes cell death in 2-ME-treated osteosarcoma cells. PMID- 18384115 TI - Microtubule assembly dynamics: an attractive target for anticancer drugs. AB - Microtubules, composed of alphabeta tubulin dimers, are dynamic polymers of eukaryotic cells. They play important roles in various cellular functions including mitosis. Microtubules exhibit differential dynamic behaviors during different phases of the cell cycle. Inhibition of the microtubule assembly dynamics causes cell cycle arrest leading to apoptosis; thus, qualifying them as important drug targets for treating several diseases including cancer, neuronal, fungal, and parasitic diseases. Although several microtubule-targeted drugs are successfully being used in cancer chemotherapy, the development of resistance against these drugs and their inherent toxicities warrant the development of new agents with improved efficacy. Several antimicrotubule agents are currently being evaluated for their possible uses in cancer chemotherapy. Benomyl, griseofulvin, and sulfonamides have been used as antifungal and antibacterial drugs. Recent reports have shown that these drugs have potent antitumor potential. These agents are shown to inhibit proliferation of different types of tumor cells and induce apoptosis by targeting microtubule assembly dynamics. However, unlike vincas and taxanes, which inhibit cancer cell proliferation in nanomolar concentration range, these agents act in micromolar range and are considered to have limited toxicities. Here, we suggest that these drugs may have a significant use in cancer chemotherapy when used in combination with other anticancer drugs. PMID- 18384116 TI - The ethical practice of psychotherapy: easily within our reach. AB - Psychotherapists confront a myriad of ethical dilemmas as they endeavor to provide effective services. This issue of the Journal of Clinical Psychology: In Session on Ethics in Psychotherapy provides psychotherapists with thoughtful reviews, case examples, and practical guidance in the major areas of ethics. Following this brief introduction, the subsequent seven articles cover Informed consent; confidentiality, privilege, and their limits; treatment of minors and their families; business matters of practice (e.g., money, fees, bartering, advertising); clinical competence and scope of practice; boundaries and nonsexual multiple relationships; and termination and abandonment. This issue is designed to promote ethical practice, to provide guidance on common ethical dilemmas, and to prevent ethical challenges before they occur. PMID- 18384117 TI - Competence and scope of practice: ethics and professional development. AB - In this article, I discuss the importance of the psychotherapist's capacities and attributes that go beyond formal education and training as they relate to both readiness for clinical training and continued competence throughout one's professional life. Professional development is essential to the maintenance of professional competence as a psychotherapist. Principles and standards from the American Psychological Association's (2002) Ethics Code are reviewed and illustrated with clinical vignettes. In striving to maintain competence, psychotherapists are strongly encouraged to focus on proactive self-care and professional development in addition to complying with the formal continuing education mandated by most states. PMID- 18384118 TI - Ethical challenges in mental health services to children and families. AB - Mental health practitioners working with children and families must attend to several ethical concerns that do not typically come into play with adult clients. The challenges for practitioners usually involve attention to four subsets of concerns that all begin with the letter c: competence, consent, confidentiality, and competing interests. Using the 4-C model, this article focuses on ethical aspects of practitioner competence, consent and assent, confidentiality, and the incongruence of interests that occurs when different people organize and set goals for psychological services. After explicating these issues, I provide recommendations for addressing them in the course of clinical practice. PMID- 18384119 TI - The CM cell line derived from liver metastasis of malignant human insulinoma is not a valid beta cell model for in vitro studies. AB - The CM cell line is derived from the ascitic fluid of a patient with liver metastasis of a malignant insulinoma. Insulin levels potentially derived from the insulinoma were detected only once in vivo in the subject with the malignant pancreatic tumor and hypoglycemia. After multiple unsuccessful attempts to detect insulin in the culture medium, insulin levels were again detected only once in vitro. In our repeated experiments, we extensively exposed early-passages of the CM cell line to 0.91 mM glucose and acutely to increasingly higher glucose concentrations (2.75, 5.5, 11, and 22 mM) and did not detect any insulin secretion as well as any significant insulin cell content. The electronic microscopic examinations of several vials containing early-passages of the CM cell lines showed a polyclonal nature of the cells mostly resembling fibroblasts. The karyotype detected severe and consistent chromosomal aberrations of the CM cell line, including the chromosome 11 tetraploidy and the genetic material translocation in three out of four chromosomes 11 at the insulin gene locus 11p15.1. These data, unfortunately, exclude the possibility of considering the CM cell line as a valid beta cell model in vitro. PMID- 18384120 TI - Can assessors and therapists predict the outcome of long-term psychotherapy in borderline personality disorder? AB - Surprisingly few studies have investigated the accuracy of prognostic assessments of therapy outcome by clinicians. The objective of the present study was to investigate the relationship between clinicians' prognostic assessments and patient characteristics and treatment outcome. Seventy-one patients with a borderline personality disorder randomly allocated to schema-focused therapy (SFT) or transference-focused psychotherapy (TFP) were assessed every 3 months for 3 years. Prognostic assessments proved to be unrelated to patients' biographical (i.e., age, gender, education level, and employment level) and clinical characteristics (i.e., number of Axis I and Axis II diagnoses, and severity of psychiatric symptoms or borderline personality pathology). Clinical assessors as well as therapists rated the probability of success for SFT to be higher than for TFP. Prospective assessments of assessors and therapists accurately predicted different indices of outcome above and independent of patient characteristics. The prediction of outcome in the TFP condition in particular proved to be valid. Identifying prognostic markers of treatment outcome as used by clinicians in their prognostic assessments may improve current prediction models and patient-treatment matching. PMID- 18384121 TI - Regarding several points of doubt of the structure of the olfactory bulb: as described by T. Blanes. AB - In order to complement the studies on the fila olfactoria completed by Dr. Santiago Ramon y Cajal, we have included a translation of "Sobre Algunos Puntos Dudosos de la Estructura del Bulbo Olfatorio" by T. Blanes, a student of Dr. Cajal. This work describes in stunning detail additional morphological aspects of the olfactory pathway, including what was at the time the modestly studied neuroglia. The neuroglia of the olfactory system has been revisited in the last several decades for its importance in the field of regenerative neuroscience. Olfactory ensheathing glia has the unique quality of providing ensheathment to neurons which traverse from the central to the peripheral nervous system and are being used as a candidate in present-day transplantation studies to mimic this phenomenon at the dorsal root entry zone after a central nervous system injury. Although this fine work has passed its centennial anniversary since initial publication, it has been widely cited throughout the years, and of recent when Pressler and Stowbridge reported Blanes cell electrophysiological recordings (Neuron V 49, 6; p 889-904, 2006). An English translation the details of what Blanes initially documented with unduplicated precision can now be made available to a wider audience in the field of neuroscience, and is especially important now that more and more present-day studies require a precise and complete understanding of the anatomical structures contained within the olfactory system. PMID- 18384122 TI - Temporal changes of astrocyte activation and glutamate transporter-1 expression in the spinal cord after spinal nerve ligation-induced neuropathic pain. AB - Astrocyte activation is involved in the neuropathic pain. As a glutamate scavenger, the glutamate transporter-1 (GLT-1) is exclusively expressed on the astrocytes and probably correlates with astrocyte activation. In the present study, we attempted to clarify the temporal changing courses of astrocyte activation and GLT-1 expression, as well as their correlations induced by a neuropathic pain model, namely, spinal nerve ligation (SNL) in which rapidly appearing (<3 days) and persistent (>21 days) mechanical allodynia and thermal hyperalgesia were presented. Immunofluorescent staining showed that GLT-1 was expressed exclusively in most (not all) of the astrocytes, even when the GLT-1 expression reached its peak. The expression of GLT-1 displayed an interesting biphasic change, with an initial up-regulation followed by a down-regulation after SNL. Our results also demonstrated that SNL induced a marked and long-term (>21 days) activation of astrocytes in the ipsilateral spinal dorsal horn. These results suggest that astrocyte activation, the change of GLT-1 expression and the potential relationship between them might play key roles in the induction and/or maintenance of neuropathic pain. The present results provide novel clues in understanding the mechanisms underlying the involvement of astrocytes and GLT-1 in the neuropathic pain. PMID- 18384123 TI - Claudin-5 is restricted to the tight junction region of uterine epithelial cells in the uterus of pregnant/gravid squamate reptiles. AB - Claudin-5, a tight junctional protein associated with ion and size selectivity, has been found in the uterus of skinks. This study has generated critical information about the molecular assembly of the tight junction at various stages of the reproductive cycle in the skink uterus. Recent studies looking at tight junctional proteins found occludin expression in the tight junction region of uterine epithelial cells in the skink uterus; however, occludin did not disclose any further information about the ions and size of ions permeating across the paracellular pathway. A approximately 22-kDa claudin-5 band was detected in the uterus of the skinks present in this study and immunohistochemistry revealed that claudin-5 redistributes to the tight junction region of the lateral plasma membrane of uterine epithelial cells in late stage pregnancy/gravidity. This finding indicates that the tight junction becomes more assembled to precisely regulate ion and solute permeation in late stage pregnancy/gravidity. Claudin-5 with its functional role as a molecular sieve due to the formation of ion and size selective pores suggests that permeation of ions smaller than 0.8 kDa are restricted when claudin-5 is redistributed to the tight junction region of the later plasma membrane. This report is the first description of the molecular mechanisms that may be involved in regulating nutrient provision in the reptilian uterus. PMID- 18384124 TI - c-Jun NH2-terminal kinase (JNK)-dependent nuclear translocation of apoptosis inducing factor (AIF) following engagement of membrane immunoglobulin on WEHI-231 B lymphoma cells. AB - WEHI-231 B lymphoma cells have been employed for analysis of antigen-induced B cell unresponsiveness because these cells undergo cell cycle arrest in G1, accompanied by induction of apoptosis. In the present study, we examined the requirement for toxic small molecules apoptosis-inducing factor (AIF) and cytochrome c, and subsequent caspase activation in apoptotic cell death in WEHI 231 and CH31 B lymphoma cells following engagement of membrane immunoglobulin (mIg). Pan-caspase inhibitor BD-fmk blocked mIg-mediated increase in cells with sub-G1 DNA content, whereas it did not affect mIg-mediated loss of mitochondrial membrane potential and phosphatidylserine exposure on B cell membrane. Dominant negative form of c-Jun NH2-terminal kinase1 (JNK1) blocked the translocation of AIF into the nuclei and cytosol from the mitochondria in the WEHI-231 and CH31 cells following mIg engagement, whereas constitutively active form of JNK1 enhanced it. This AIF translocation was also blocked by Bcl-xL, but not by BD fmk. Moreover, AIF-deficient clones via small interfering RNA (siRNA)-mediated method showed small increase in loss of mitochondrial membrane potential. After mIg engagement, the AIF-deficient clones displayed an enhanced sensitivity to mIg mediated apoptosis, concomitant with translocation of a residual AIF into the nuclei, compared with control clone. Our findings are compatible with the notion that AIF has dual role, with a proapoptotic function in the nuclei and a distinct anti-apoptotic function in the mitochondria. These observations would be valuable for analysis of B cell unresponsiveness and hopefully for treatment of diseases involving B cell dysfunction. PMID- 18384125 TI - Baicalein stimulates osteoblast differentiation via coordinating activation of MAP kinases and transcription factors. AB - The identification of anabolic agents that directly stimulate bone formation has recently attracted greater interest. Here, baicalein was identified as a natural compound that stimulates the differentiation of mouse osteoblastic MC3T3-E1 subclone 4 cells. Baicalein induced the activation of NF-kappaB in the initiation stage of osteoblast differentiation, and it activated the MAP kinase/NF-kappaB signaling pathway and induced the expression of osteoblast differentiation markers in the early stage. In the late stage, baicalein stimulated the calcium deposition with the activation of MAP kinases and AP-1 family members such as Fra 1 and Fra-2. Another transcription factor, NFATc1, was slightly induced by baicalein in the late stage. Thus, baicalein could stimulate the osteoblast differentiation via the activation of complexly coordinated signaling pathways that include MAP kinases and transcription factors such as NF-kappaB, AP-1, and NFATc1. PMID- 18384126 TI - Inhibition of adipocyte differentiation by phytoestrogen genistein through a potential downregulation of extracellular signal-regulated kinases 1/2 activity. AB - In the current study, we investigated the effects of genistein on adipogenic differentiation of mouse bone marrow-derived mesenchymal stem cell (BMSC) cultures and its potential signaling pathway. The terminal adipogenic differentiation was assessed by western-blotting analysis of adipogenic-specific proteins such as PPARgamma, C/EBPalpha, and aP2 and the formation of adipocytes. Treatment of mouse BMSC cultures with adipogenic cocktail resulted in sustained activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), which are members of the mitogen-activated protein kinase (MAPK) family, at the early phase of adipogenesis (from days 3 to 9). Inhibition of ERK1/2 activation by PD98059, a specific MEK inhibitor, reversed the induced adipogenic differentiation. Genistein dose-dependently decreased the phosphorylation of ERK1/2 in mouse BMSC cultures. Genistein incubation for the entire culture period, as well as that applied during the early phase of the culture period, significantly inhibited the adipogenic differentiation of mouse BMSC cultures. While genistein was incubated at the late stage (after day 9), no inhibitory effect on adipogenic differentiation was observed. BMSC cultures treated with genistein in the presence of fibroblast growth factor-2 (FGF-2), an activator of the ERK1/2 signaling pathway, expressed normal levels of ERK1/2 activity, and, in so doing, are capable of undergoing adipogenesis. Our results suggest that activation of the ERK1/2 signaling pathway during the early phase of adipogenesis (from days 3 to 9) is essential to adipogenic differentiation of BMSC cultures, and that genistein inhibits the adipogenic differentiation through a potential downregulation of ERK1/2 activity at this early phase of adipogenesis. PMID- 18384127 TI - Spatiotemporal dynamics of regulatory protein recruitment at DNA damage sites. AB - Mammalian cells are constantly threatened by multiple types of DNA lesions arising from various sources like irradiation, environmental agents, replication errors or by-products of the normal cellular metabolism. If not readily detected and repaired these lesions can lead to cell death or to the transformation of cells giving rise to life-threatening diseases like cancer. Multiple specialized repair pathways have evolved to preserve the genetic integrity of a cell. The increasing number of DNA damage sensors, checkpoint regulators, and repair factors identified in the numerous interconnected repair pathways raises the question of how DNA repair is coordinated. In the last decade, various methods have been developed that allow the induction of DNA lesions and subsequent real time analysis of repair factor assembly at DNA repair sites in living cells. This combination of biophysical and molecular cell biology methods has yielded interesting new insights into the order and kinetics of protein recruitment and identified regulatory sequences and selective loading platforms for the efficient restoration of the genetic and epigenetic integrity of mammalian cells. PMID- 18384128 TI - Youth organizing: From youth development to school reform. AB - Over the past twenty years, youth organizing has grown across the country. Through organizing, young people identify issues of concern and mobilize their peers to build action campaigns to achieve their objectives. Youth organizing has been appreciated for its contributions to youth and community development. The authors use two case studies to trace the more recent emergence of youth organizing as an important force for school reform. The Boston-based Hyde Square Task Force began with a focus on afterschool programming, but its youth leaders now organize to get Boston Public Schools to adopt a curriculum addressing sexual harassment. Meanwhile, the Baltimore Algebra Project began as a peer-to-peer tutoring program but now also organizes to demand greater funding for Baltimore schools. These cases illustrate a broader phenomenon where students reverse the deficit paradigm by acting out of their own self-interest to become agents of institutional change. PMID- 18384129 TI - Inhibition of osteoblastic metalloproteinases in mice prevents bone loss induced by oestrogen deficiency. AB - Matrix metalloproteinases (MMPs) are key mediators in extra-cellular matrix remodelling and implicated primarily in bone growth, and particularly in osteoclastic bone resorption. We hypothesise that MMPs have a role in the increased bone remodelling resulting from oestrogen deficiency. Transgenic (TG) mice overexpressing TIMP-1 in their osteoblastic cells and their wild-type (WT) littermates were ovariectomised. One month after surgery, bone mineral density (BMD) and bone microarchitecture were assessed. Primary cells from WT and TG mice were used to determine how TIMP-1 affects osteoclast and osteoblastic cells. The reduction of BMD induced by ovariectomy in WT mice was not observed in the transgenic mice. The transgene overexpression also dampened the post-ovariectomy increase in bone resorption in contrast to the WT mice. In vivo, osteoclastic surfaces and D-pyridinoline were not increased in TG mice, and ex vivo, the differentiation of osteoclasts from TG bone marrow precursor cells were unaffected by in vivo oestrogen deficiency or treatment. We showed also that TIMP 1 overexpression reduces and delays the osteoblastic proliferation and differentiation respectively, and reduced the generation of the active form of TGFbeta1 in the supernatant of TG osteoblasts. Our findings support the hypothesis that in vivo inhibition of osteoblastic MMPs prevented the bone loss induced by oestrogen deficiency, with a significant decrease in bone resorption. This effect was presumably resulting from (1) a direct inhibition of osteoclastic resorption activity by the TIMP-1 and (2) the modification in the local activation of extra-cellular signalling factors such as TGFbeta1 and the OPG/RANKL ratio. PMID- 18384130 TI - Concomitant activation of Wnt pathway and loss of mismatch repair function in human melanoma. AB - Constitutive activation of the Wnt pathway plays a key role in the development of colorectal cancer and has also been implicated in the pathogenesis of other malignancies. Deregulation of Wnt signaling mainly occurs through genetic alterations of APC, the beta-catenin gene (CTNNB1), AXIN1 and AXIN2, leading to stabilization of beta-catenin. Physiologically, AXIN2 is transcriptionally induced on Wnt signaling activation and acts as a negative feedback regulator of the pathway. In colorectal cancer, mutations in CTNNB1 and AXIN2 occur preferentially in tumors with inactivation of the mismatch repair (MMR) genes MSH2, MLH1, or PMS2. In this study, the expression of beta-catenin and AXIN2, and the mutational status of CTNNB1, APC, and AXIN2 were evaluated in two MMR deficient (PR-Mel and MR-Mel) and seven MMR-proficient human melanoma cell lines. Only PR-Mel and MR-Mel cells showed nuclear accumulation of beta-catenin and expression of the AXIN2 gene, and hence, constitutive activation of Wnt signaling. Mutational analysis identified a somatic heterozygous missense mutation in CTNNB1 exon three and a germline heterozygous deletion within AXIN2 exon seven in PR-Mel cells, and a somatic biallelic deletion within APC in MR-Mel cells. Deregulation of Wnt signaling and a defective MMR system were also present in the original tumor of PR and MR patients. Thus, we describe additional melanomas with mutations in CTNNB1 and APC, identify for the first time a germline AXIN2 mutation in a melanoma patient and suggest that inactivation of the MMR system and deregulation of the Wnt/beta-catenin signaling pathway cooperate to promote melanoma development and/or progression. PMID- 18384131 TI - Identification of genomic changes associated with cisplatin resistance in testicular germ cell tumor cell lines. AB - Since the introduction of cisplatin into the clinic, the treatment of patients with a variety of solid tumors including testicular germ cell tumors, ovarian and lung cancers, has dramatically improved. One of the main causes for therapeutic failure in these malignancies is the development of drug resistance. Testicular germ cell tumors (TGCTs), the most common malignancy in young men, exhibit extreme sensitivity to cisplatin-based chemotherapy, making them an ideal model for investigating the mechanisms of cisplatin chemo-sensitivity and resistance. TGCT development and pathogenesis have been well studied but little is known about the genetic background in chemo-resistant cases. We investigated genomic differences between three TGCT parental cell lines and their cisplatin resistant derivatives. Using 10K single nucleotide polymorphism (SNP) microarray analysis, we identified two small chromosomal regions with consistent copy number changes across all three pairs of resistant cell lines. These were an 8.7 Mb region at 6q26-27, which displayed consistent copy number gain and a 0.3 Mb deletion involving 4 SNPs at 10p14. Both the chromosomal gain and loss were confirmed by fluorescence in situ hybridization. The significance of these regions should be further investigated as they may contain key genes involved in the development of chemo- resistance to cisplatin-based treatment in TGCTs and other cancers. PMID- 18384132 TI - Thirty years of advocacy in San Francisco: lessons learned and the next generation of leadership. AB - Professional advocacy organizations are often challenged by the question of their authentic community representation and their ability to balance short-term pragmatism with strategic plans for long-term, systemic change. Coleman Advocates, one of the nation's most effective child advocacy organizations, has taken up this challenge under the leadership of a next-generation leader of color who followed a dynamic director of the baby boom generation. In this piece, Coleman's thirty years of social change strategies are analyzed from the perspective of this new executive director, who has facilitated the latest organizational shift that deepens its commitment to building bottom-up grassroots leadership and community power while keeping the best of the professional, staff led advocacy model. Issues of race, accountability, power, and movement building are addressed through the lens of one organization's evolution, with the goal of building a long-term movement that will achieve racial and economic equity for all children and families. PMID- 18384133 TI - Faith-based organizing for youth: one organization's district campaign for small schools policy. AB - Oakland Community Organizations (OCO) has worked for over ten years to improve educational opportunities in low-income neighborhoods in Oakland, California. The work of thousands of parent, teacher, youth, and community leaders has resulted in the formation of nearly fifty new small schools and more than ten charters, creating settings for individualized learning environments and the opportunity for quality choices for many of Oakland's low-income families. In this article, OCO's executive director, Ron Snyder, outlines a four-phase organizing process undertaken by OCO, based on a set of organizing principles that have sustained community-led education reform despite constant changes in the political and school district environment: the centrality of love (self-interest) as a motivator for advocacy; the importance of quality research and powerful ideas (vision) as alternatives to the status quo; application of a model that creates a common structure, language, and experience to sustain leaders; the need for institutional and network power to apply leverage; the flexibility to seize opportunity when the window is open; and faithfulness to the object of our love: our children. PMID- 18384134 TI - Developing the field of youth organizing and advocacy: what foundations can do. AB - For more than a decade, the Evelyn and Walter Haas, Jr. Fund has seeded many San Francisco Bay Area youth organizing and advocacy programs. Now that the field is maturing, argues the fund's vice president of programs, foundations have a critical programmatic and capacity-building role to play. The author offers analysis and strategies for integrating youth development grant making across foundation interest areas. The programs described illustrate the diversity of youth organizing and advocacy programs that could be supported by funders, whether or not any particular philanthropic institution has a grant-making focus on youth development or youth organizing. The article ends with an in-depth portrait of the self-reported needs of youth organizing and advocacy programs and concrete strategies for foundations seeking to more effectively enable youth organizing and advocacy to play an important role in bringing about a more vibrant, diverse, and effective democratic culture. PMID- 18384135 TI - Community organizing and youth advocacy. PMID- 18384136 TI - Organizations advocating for youth: the local advantage. AB - Youth occupy a unique place in our democratic society. They must primarily rely on others to speak on their behalf as decisions are made about the allocation of resources within and across various youth-serving institutions. Advocacy organizations comprise crucial representational assets for all youth, but America's poorest children and youth especially need an effective voice to speak for and about them. Yet advocates for youth in urban areas face tough challenges since urban voters typically have few positive connections to youth. This article draws on three years of research focused on three organizations in the San Francisco Bay Area that have successfully advocated for better policies for youth. The authors explore the strategies that these organizations have employed to overcome the challenges they face, with particular attention to the advantages that follow from advocating at the local rather than at the state or federal level. PMID- 18384137 TI - The development of collective moral leadership among parents through education organizing. AB - It is often assumed that parent participation in schools is primarily based on self-interest and that this is a frequent source of contention between parents and teachers. This article examines the experiences of members of the Jamaica Plain Parent Organizing Project (JP-POP), a community-based organization in Boston, and reveals that some parents have learned to act beyond their individual self-interest and to organize on behalf of the entire community as a result of their participation. The authors present qualitative data from interviews with JP POP members to ascertain the motivations behind their initial decisions to become involved in education, the benefits they derive from their participation, and the gradual evolution of narrow definitions of self-interest to more communal understandings. Finally, they draw out implications for the potential capacity enhancement of community-based organizations in education at both the institutional and district levels. PMID- 18384139 TI - Negotiating reform: young people's leadership in the educational arena. AB - Youth organizing within the institutional context of community-based organizations has grown exponentially. Drawing on interviews with more than eighty organizers, youth, and educators, this article examines young people's experiences as they organize to expand educational opportunities for themselves and their peers in urban school districts. The authors explore educator responses to youth organizing and analyze how race- and class-based assumptions about youth leadership, as well as differing cultural norms between schools and youth organizing groups, pose challenges for young people fighting for education reform. The authors describe three strategies youth organizing groups use to address these challenges: intensive leadership development, targeted relationship building with district administrators, and alliance building. Implications for both educators and youth organizing groups are discussed. PMID- 18384140 TI - Oxidation and nuclear localization of thioredoxin-1 in sparse cell cultures. AB - Reactive oxygen species (ROS) were once viewed only as mediators of toxicity, but it is now recognized that they also contribute to redox signaling through oxidation of specific cysteine thiols on regulatory proteins. Cells in sparse cultures have increased ROS relative to confluent cultures, but it is not known whether protein redox states are affected under these conditions. The purpose of the present study was to determine whether culture conditions affect the redox state of thioredoxin-1 (Trx1), the protein responsible for reducing most oxidized proteins in the cytoplasm and nucleus. The results showed that Trx1 was more oxidized in sparse HeLa cell cultures than in confluent cells. The glutathione pool was also more oxidized, demonstrating that both of the major cellular redox regulating systems were affected by culture density. In addition, the total amount of Trx1 protein was lower and the subcellular distribution of Trx1 was different in sparse cells. Trx1 in sparse cultures was predominantly nuclear whereas it was predominantly cytoplasmic in confluent cultures. This localization pattern was not unique to HeLa cells as it was also observed in A549, Cos-1 and HEK293 cells. These findings demonstrate that Trx1 is subject to changes in expression, redox state and subcellular localization with changing culture density, indicating that the redox environments of the cytoplasm and the nucleus are distinct and have different requirements under different culture conditions. PMID- 18384141 TI - Nephrocalcinosis and disordered calcium metabolism in two children with SHORT syndrome. AB - Reports of SHORT syndrome have, to date, focused on the clinical features which lie at the core of the diagnosis but there has been little by way of report of long-term outcome, either in terms of medical complications or of intellectual development. We now report two children in whom nephrocalcinosis has developed and a third, adult, with similar findings. It may be that clinicians should be looking among cases of unexplained hypercalcaemia for an alternative phenotypic presentation of this short stature syndrome. PMID- 18384142 TI - Unrelated chromosomal anomalies found in patients with suspected 22q11.2 deletion. AB - Screening for 22q11.2 deletions has not an easy approach due to the wide variability of their associated phenotype. Many clinical features overlap with those of other known syndromes and reported loci. Patients referred to exclude a 22q11.2 deletion are usually tested with a locus-specific FISH probe, with 10% positive cases depending on the selection criteria, but patients testing negative for FISH at 22q11.2 may have other chromosomal aberrations in routine cytogenetic analysis. We tested 819 patients suspected of having a 22q11.2 deletion. Eighty eight patients (10.7%) were positive for 22q11.2 deletion, whereas 30 patients (3.7%) showed other chromosomal abnormalities involving deletions and duplications, derivative chromosomes, marker chromosomes, apparently balanced and unbalanced translocations and sex chromosome aneuploidies. Of these alterations, 28 did not involve region 22q11 and most had not been associated with 22q11.2 deletion phenotype before. We discuss the similarity of DiGeorge/velocardiofacial syndrome with other known clinical entities and suggest correlations between the new loci and the observed clinical features. The frequency of unrelated chromosomal anomalies reported in this study and in other previous reports highlights the importance of conventional cytogenetic analysis as an initial genome-wide screening tool in all referred patients, and provides useful data to optimize diagnostic and screening protocols according to the most frequent chromosomal findings. PMID- 18384143 TI - A prenatally ascertained X;Y translocation characterized using conventional and molecular cytogenetics. PMID- 18384144 TI - A new candidate locus for bilateral perisylvian polymicrogyria mapped on chromosome Xq27. AB - Polymicrogyria (PMG) is characterized by an excessive number of small and prominent brain gyri, separated by shallow sulci. Bilateral perisylvian polymicrogyria (BPP) is the most common form of PMG. Clinical signs include pseudobulbar paresis, mental retardation, and epilepsy. Familial forms of BPP have been described and a candidate locus was previously mapped to chromosome Xq28, distal do marker DXS8103. The objective of this study was to perform linkage analysis in one family segregating BPP. A total of 15 individuals, including 8 affected patients with BPP were evaluated. Family members were examined by a neurologist and subjected to magnetic resonance imaging scans. Individuals were genotyped for 18 microsatellite markers, flanking a 42.3 cM interval on ch Xq27-q28. Two-point and multipoint linkage analysis was performed using the LINKAGE package and haplotype reconstruction was performed by GENEHUNTER software. Our results showed a wide spectrum of clinical manifestations in affected individuals with BPP, ranging from normal to mild neurological abnormalities. Two-point linkage analysis yield a Zmax = 2.06 at theta = 0.00 for markers DXS1205 and DXS1227. Multipoint lod-scores indicate a candidate interval of 13 cM between markers DSXS1205 and DXS8043, on ch Xq27.2 Xq27.3. These results point to a new locus for BPP in a more centromeric location than previously reported. PMID- 18384145 TI - Two new cases of pure 1q terminal deletion presenting with brain malformations. AB - We describe two new cases of pure 1q terminal deletions. BAC FISH analysis precisely defined the size of deletions. The first is a girl with 10.3-Mb deletion showed typical features of 1q43 deletion as well as a simplified gyral pattern, which was rarely found in 1q43 deletion. The other boy also presented with most of 1q43 deletion features but several atypical symptoms were noted including hydrocephalus, adducted thumbs, and flexion restriction of proximal interphalangeal joints in left hand. A concomitant novel missense mutation in L1CAM was identified in addition to 11.5-Mb deletion. Reviewing all the cases of pure 1q terminal deletion in the literature suggests that it is a clinically recognizable syndrome. PMID- 18384146 TI - Trisomy 20q caused by interstitial duplication 20q13.2: clinical report and literature review. AB - We report on a 3-year-old boy with moderate developmental delay, abnormal craniofacial features and ventricular septal defect resulting from trisomy of the long arm of chromosome 20. The cytogenetic defect consists of a de novo isolated interstitial duplication in distal 20q [dup(20)(q13.2q13.2)]. The duplication was detected by comparative genomic hybridization (CGH) and confirmed by array CGH. Other cases of comparable trisomies are reviewed. This new case further delineates the recognizable phenotype of trisomy 20q13 --> 20qter and highlights the relevance of CGH for the detection of such rearrangements. PMID- 18384147 TI - Refinement of noncalorimetric determination of the change in heat capacity, DeltaC(p), of protein unfolding and validation across a wide temperature range. AB - The change in heat capacity, DeltaC(p), on protein unfolding has been usually determined by calorimetry. A noncalorimetric method which employs the Gibbs Helmholtz relationship to determine DeltaC(p) has seen some use. Generally, in this method the free energy change on unfolding of the protein is determined at a variety of temperatures and the temperature at which DeltaG is zero, T(m), and change in enthalpy at T(m) are determined by thermal denaturation and DeltaC(p) is then calculated using the Gibbs-Helmholtz equation. We show here that an abbreviated method with stability determinations at just two temperatures gives values of DeltaC(p) consistent with values from free energy change on unfolding determination at a much wider range of temperatures. Further, even the free energy change on unfolding from a single solvent denaturation at the proper temperature, when coupled with the melting temperature, T(m), and the van't Hoff enthalpy, DeltaH(vH), from a thermal denaturation, gives a reasonable estimate of DeltaC(p), albeit with greater uncertainty than solvent denaturations at two temperatures. We also find that nonlinear regression of the Gibbs-Helmholtz equation as a function of stability and temperature while simultaneously fitting DeltaC(p), T(m), and DeltaH(vH) gives values for the last two parameters that are in excellent agreement with experimental values. PMID- 18384148 TI - Region-specific role of water in collagen unwinding and assembly. AB - Conformational stability of the collagen triple helix affects its turnover and determines tissue homeostasis. Although it is known that the presence of imino acids (prolines or hydroxyprolines) confer stability to the molecule, little is known regarding the stability of the imino-poor region lacking imino acids, which plays a key role in collagen cleavage. In particular, there have been continuing debates about the role of water in collagen stability. We addressed these issues using molecular dynamics simulations on 30-residue long collagen triple helices, including a structure that has a biologically relevant 9-residue imino-poor region from type III collagen (PDB ID: 1BKV). A torsional map approach was used to characterize the conformational motion of the molecule that differ between imino-rich and imino-poor regions. At temperatures 300 K and above, unwinding initiates at a common cleavage site, the glycine-isoleucine bond in the imino poor region. This provides a linkage between previous observations that unwinding of the imino-poor region is a requirement for collagenase cleavage, and that isolated collagen molecules are unstable at body temperature. We found that unwinding of the imino-poor region is controlled by dynamic water bridges between backbone atoms with average lifetimes on the order of a few picoseconds, as the degree of unwinding strongly correlated with the loss of water bridges, and unwinding could be either prevented or enhanced, respectively by enforcing or forbidding water bridge formation. While individual water bridges were short lived in the imino-poor region, the hydration shell surrounding the entire molecule was stable even at 330 K. The diameter of the hydrated collagen including the first hydration shell was about 14 A, in good agreement with the experimentally measured inter-collagen distances. These results elucidate the general role of water in collagen turnover: water not only affects collagen cleavage by controlling its torsional motion, but it also forms a larger-scale lubrication layer mediating collagen self-assembly. PMID- 18384149 TI - Temperature-dependent downhill unfolding of ubiquitin. II. Modeling the free energy surface. AB - To provide evidence for the interpretation of temperature-dependent unfolding kinetics and the downhill unfolding scenario presented in the accompanying experimental article (Part I), the free energy surface of ubiquitin unfolding is calculated using statistical mechanical models of the Munoz-Eaton (ME) form. The models allow only two states for each amino acid residue, folded or unfolded, and permutations of these states generate an ensemble of microstates. One-dimensional free energy curves are calculated using the number of folded residues as a reaction coordinate. The proposed sequential unfolding of ubiquitin's beta-sheet is tested by mapping the free energy onto two reaction coordinates inspired by the experiment as follows: the number of folded residues in ubiquitin's stable beta-strands I and II and those of the less stable strands III-V. Although the original ME model successfully captures folding features of zipper-like one dimensional folders, it misses important tertiary interactions between residues that are far from each other in primary sequence. To take tertiary contacts into account, partially folded microstates based on a spherical growth model are included in the calculation and compared with the original model. By calculating the folding probability of each residue for a given point on the free energy surface, the unfolding pathway of ubiquitin is visualized. At low temperature, thermal unfolding occurs along a sequential unfolding pathway as follows: disruption of the beta-strands III-V followed by unfolding of the strands I and II. At high temperature, multiple unfolding routes are formed. The heterogeneity of the transition state explains the global nonexponential unfolding observed in the T-jump experiment at high temperature. The calculation also reports a high stability for the alpha-helix of ubiquitin. PMID- 18384150 TI - Structure determination of Discoidin II from Dictyostelium discoideum and carbohydrate binding properties of the lectin domain. AB - The social amoeba Dictyostelium discoideum adopts a cohesive stage upon starvation and then produces Discoidin I and II, two proteins able to bind galactose and N-acetyl-galactosamine. The N-terminal domain or discoidin domain (DS) is widely distributed in eukaryotes where it plays a role in extracellular matrix binding while the C-terminal domain displays sequence similarities to invertebrate lectins. We present the first X-ray structures of the wild-type and recombinant Discoidin II in unliganded state and in complex with monosaccharides. The protein forms a homotrimer which presents two binding surfaces situated on the opposite boundaries of the structure. The binding sites of the N-terminal domain contain PEG molecules that could mimics binding of natural ligand. The C terminal lectin domain interactions with N-acetyl-D-galactosamine and methyl-beta galactoside are described. The carbohydrate binding sites are located at the interface between monomers. Specificity for galacto configuration can be rationalized since the axial O4 hydroxyl group is involved in several hydrogen bonds with protein side chains. Titration microcalorimetry allowed characterization of affinity and demonstrated the enthalpy-driven character of the interaction. Those results highlight the structural differentiation of the DS domain involved in many cell-adhesion processes from the lectin activity of Dictyostelium discoidins. PMID- 18384151 TI - Temperature-dependent downhill unfolding of ubiquitin. I. Nanosecond-to millisecond resolved nonlinear infrared spectroscopy. AB - Transient thermal unfolding of ubiquitin is investigated using nonlinear infrared spectroscopy after a nanosecond laser temperature jump (T-jump). The abrupt change in the unfolding free energy surface and the ns time resolution allow us to observe a fast response on ns to micros time-scales, which we attribute to downhill unfolding, before a cross-over to ms kinetics. The downhill unfolding by a sub-population of folded proteins is induced through a shift of the barrier toward the native state. By adjusting the T-jump width, the effect of the initial (T(i)) and final (T(f)) temperature on the unfolding dynamics can be separated. From the amplitude of the fast downhill unfolding, the fractional population prepared at the unfolding transition state is obtained. This population increases with both T(i) and with T(f). A two-state kinetic analysis of the ms refolding provides thermodynamic information about the barrier height. By a combination of the fast and slow unfolding and folding parameters, a quasi-two-state kinetic analysis is performed to calculate the time-dependent population changes of the folded state. This calculation coincides with the experimentally obtained population changes at low temperature but deviations are found in the T-jump from 67 to 78 degrees C. Using temperature-dependent barrier height changes, a temperature Phi value analysis is performed. The result shows a decreasing trend of Phi(T) with temperature, which indicates an increase of the heterogeneity of the transition state. We conclude that ubiquitin unfolds along a well-defined pathway at low temperature which expands with increasing temperature to include multiple routes. PMID- 18384152 TI - Structure-based tailoring of compound libraries for high-throughput screening: discovery of novel EphB4 kinase inhibitors. AB - High-throughput docking is a computational tool frequently used to discover small molecule inhibitors of enzymes or receptors of known three-dimensional structure. Because of the large number of molecules in chemical libraries, automatic procedures to prune multimillion compound collections are useful for high throughput docking and necessary for in vitro screening. Here, we propose an anchor-based library tailoring approach (termed ALTA) to focus a chemical library by docking and prioritizing molecular fragments according to their binding energy which includes continuum electrostatics solvation. In principle, ALTA does not require prior knowledge of known inhibitors, but receptor-based pharmacophore information (hydrogen bonds with the hinge region) is additionally used here to identify molecules with optimal anchor fragments for the ATP-binding site of the EphB4 receptor tyrosine kinase. The 21,418 molecules of the focused library (from an initial collection of about 730,000) are docked into EphB4 and ranked by force field-based energy including electrostatic solvation. Among the 43 compounds tested in vitro, eight molecules originating from two different anchors show low micromolar activity in a fluorescence-based enzymatic assay. Four of them are active in a cell-based assay and are potential anti-angiogenic compounds. PMID- 18384153 TI - Nanoparticulate biopolymers deliver insulin orally eliciting pharmacological response. AB - The aim of this study was to characterize and evaluate a novel oral insulin nanoparticulate system based on alginate-dextran sulfate core, complexed with a chitosan-polyethylene glycol-albumin shell. Insulin-loaded nanospheres (25, 50, 100 IU/kg) administered orally to diabetic rats reduced glycemia in a dose dependent manner. This effect lasted over 24 h with a maximal effect after 14 h. Nanospheres increased insulin plasma level and improved glycemic response to an oral glucose overload. After 4 days oral administration (50 IU/kg/day), the metabolic status of diabetic rats improved with a reduction in water intake, urine excretion and proteinuria. FITC-insulin-loaded nanospheres administered to an isolated intestinal loop were taken up by the intestinal mucosa. They strongly adhered to villus apical enterocytes and markedly labeled Peyer's patches. It is concluded that nanospheres preserve insulin and exert an antidiabetic effect after oral administration. This is explained by a protective effect against proteolytic enzymes by the albumin coating, by the mucoadhesive properties of chitosan-polyethylene glycol, and by the possibility of chitosan reversibly altering tight junctions leading to an improved absorption of insulin. This formulation demonstrates beneficial effects on diabetic symptoms and will be of interest in the treatment of diabetes with oral insulin. PMID- 18384154 TI - Effect of dimethyl-beta-cyclodextrin concentrations on the pulmonary delivery of recombinant human growth hormone dry powder in rats. AB - The aim of this article is to prepare and characterize inhalable dry powders of recombinant human growth hormone (rhGH), and assess their efficacy for systemic delivery of the protein in rats. The powders were prepared by spray drying using dimethyl-beta-cyclodextrin (DMbetaCD) at different molar ratios in the initial feeds. Size exclusive chromatography was performed in order to determine protecting effect of DMbetaCD on the rhGH aggregation during spray drying. By increasing the concentration of DMbetaCD, rhGH aggregation was decreased from 9.67 (in the absence of DMbetaCD) to 0.84% (using DMbetaCD at 1000 molar ratio in the spray solution). The aerosol performance of the spray dried (SD) powders was evaluated using Andersen cascade impactor. Fine particle fraction values of 53.49%, 33.40%, and 23.23% were obtained using DMbetaCD at 10, 100, and 1000 molar ratio, respectively. In vivo studies showed the absolute bioavailability of 25.38%, 76.52%, and 63.97% after intratracheal insufflation of the powders produced after spray drying of the solutions containing DMbetaCD at 10, 100, and 1000 molar ratio, respectively in rat. In conclusion, appropriate cyclodextrin concentration was achieved considering the protein aggregation and aerosol performance of the SD powders and the systemic absorption following administration through the rat lung. PMID- 18384155 TI - Pharmaceutical aspects of drug eluting stents. AB - This review focuses on the pharmaceutical aspects of the development of drug eluting stents. It discusses the different processes that can be used to obtain a controlled release of a drug from the stent as well as the coatings therefore applied. Results obtained for stents already available on the market or in a far stage of development are discussed. In a final part possible future research areas as well as expected new evolutions in the design of drug eluting stents are presented. PMID- 18384156 TI - Pharmacokinetics of P-glycoprotein inhibition in the rat blood-brain barrier. AB - This article describes the experimental set-up and pharmacokinetic modeling of P glycoprotein function in the rat blood-brain barrier using [(11)C]verapamil as the substrate and cyclosporin A as an inhibitor of P-gp. [(11)C]verapamil was administered to rats as an i.v. bolus dose followed by graded infusions to obtain steady-state concentrations in the brain during 70 min. CsA was administered as a bolus followed by a constant infusion 20 min after the start of the [(11)C]verapamil infusion. The brain uptake of [(11)C]verapamil over 2 h was portrayed in a sequence of PET scans in parallel with measurement of [(11)C]verapamil concentrations in blood and plasma and CsA concentrations in blood. Mixed effects modeling in NONMEM was used to build a pharmacokinetic model of CsA-induced P-gp inhibition. The brain pharmacokinetics of [(11)C]verapamil was well described by a two-compartment model. The effect of CsA on the uptake of [(11)C]verapamil in the brain was best described by an inhibitory indirect effect model with an effect on the transport of [(11)C]verapamil out of the brain. The CsA concentration required to obtain 50% of the maximal inhibition was 4.9 microg/mL (4.1 microM). The model parameters indicated that 93% of the outward transport of [(11)C]verapamil was P-gp mediated. PMID- 18384157 TI - Binge drinking and violence against intimate partners in Russia. AB - This study is the first to provide information on the association between intimate partner violence (IPV) and binge drinking among Russian university students. Using data from 500 (58% female) university students from the four Russian sites of the International Dating Violence Study, we found gender differences in rates of IPV perpetration and in the association between binge drinking and IPV. Specifically, more females than males perpetrated IPV, and the associations between binge drinking and IPV were stronger for the female students than for the male students. In addition, antisocial traits and behavior (ATSB) were significantly related to both binge drinking and IPV perpetration for males and females. For males, the relatively weak associations between binge drinking and IPV perpetration disappeared once ASTB was accounted for. For females, the relationship decreased but remained significant when ATSB was statistically controlled. Path analyses confirmed that this pattern of relationships would be consistent with ATSB serving as a partial mediator between binge drinking and IPV perpetration. However, other alternative mediation and moderation models for the relationships between binge drinking, IPV perpetration, and ATSB could not be ruled out with this one-wave correlational study. PMID- 18384158 TI - Preparation and cytocompatibility of PLGA scaffolds with controllable fiber morphology and diameter using electrospinning method. AB - Poly(lactide-co-glycolide) (PLGA) scaffolds prepared by the electrospinning technology were paid more attention in tissue engineering, but detailed investigation on the influence of process parameters on scaffold morphology and fiber diameters as well as distribution remains to be conducted. In the current study, PLGA concentration, electric field strength, and feeding rate were varied to prepare different PLGA scaffolds. It was shown that with the varying of the above parameters, the electrospun porous PLGA scaffolds exhibited fibrous morphology either with/out beads or bead-fiber string. The diameter of nanofibers increased with the PLGA concentration and feeding rate, whereas the electric field strength exerted only minor effects on the average diameter of the nanofibers. The cytocompatibility of the scaffold prepared from optimizing fabrication parameters was evaluated using human fibroblasts. Good cell attachment, fast proliferation rate, characteristic fibroblast morphology with homogeneous distribution was observed on the scaffold. As a whole, this study demonstrated that the fiber morphology and diameter of the electrospun PLGA scaffolds could be tailored by controlling fabrication parameters. And nanofibrous PLGA could act as a biologically preferred scaffold for fibroblasts growth with maintenance of characteristic morphology. PMID- 18384159 TI - Regulation of adult human mesenchymal stem cells into osteogenic and chondrogenic lineages by different bioreactor systems. AB - The aim of this study was to examine the feasibility of expanding and regulating mesenchymal stem cells (MSCs) from isolated adult human bone marrow mononuclear cells, seeded on gelatin-hyaluronic acid biomatrices, and then to quantitatively compare the gene expression in three different culture systems. Individual and interactive effects of model system parameters on construct structure, function, and molecular properties were evaluated. The results showed that these adult human MSCs even at old age not only expressed primitive mesenchymal cell markers but also maintained a high level of colony-forming efficiency and were capable of differentiating into osteoblasts, chondrocytes, and adipocytes upon appropriate inductions. After 21 days of culture, we found that the osteoblastic and chondrocytic lineage gene expression were earlier and higher expressed in spinner flask bioreactor culture group when compared with the static culture and rotating wall vessel reactor culture. The osteogenic lineage proteins type I collagen, alkaline phosphatase, and osteocalcin were strongly stained in histological sections of spinner flask bioreactor culture, whereas these were less detected in the other two groups, especially in rotating wall vessel reactor culture. As for the markers associated with the chondrogenic lineage differentiation proteins, type II collagen was apparently expressed in spinner flask culture group, while the expression of proteoglycans (aggreacan, decorin) in three culture conditions took the lead of each other. We conclude that the spinner flask bioreactor with appropriate induction medium reported in this study may be used to rapidly expand adult MSCs and is likely to possess better induction results toward osteoblastic and chondrocytic lineages. PMID- 18384160 TI - Reinforcement of bone cement using zirconia fibers with and without acrylic coating. AB - Acrylic (polymethylmethacrylate or PMMA) bone cement was modified by the addition of high-strength zirconia fibers with average lengths of 200 microm and diameters of 15 microm or 30 microm. A novel emulsion polymerization process was developed to encapsulate individual fibers in PMMA. Improvements in tensile and compressive properties as well as in fracture toughness were investigated upon incorporation of uncoated and acrylic coated zirconia fibers. Bone cements were reinforced with 2% by volume of the 15 microm diameter and 5% by volume of the 30 microm fibers. Results indicate that elastic modulus and ultimate strength of bone cements reinforced with zirconia fibers were higher than controls, being the largest for cements reinforced with 30 microm diameter fibers. The fracture toughness of the cement increased by 23% and 41% by the addition of 15 microm and 30 microm fibers, respectively. Coating of individual zirconia fibers did not result in improved material properties of bone cements. The use of uncoated or acrylic coated 30 microm fibers is recommended based on the significant increases in ultimate strength and fracture toughness of the cements. PMID- 18384161 TI - Nanocomposite Ti/hydrocarbon plasma polymer films from reactive magnetron sputtering as growth support for osteoblast-like and endothelial cells. AB - Nanocomposite Ti/hydrocarbon plasma polymer (Ti/ppCH) films were deposited by DC magnetron sputtering of titanium target in n-hexane, argon, or a mixture of these two gases. The resultant films were heterogeneous, with inorganic regions of nanometer scale distributed within a plasma polymer matrix. The titanium content was controlled by adjusting the argon/n-hexane ratio in the working gas. In the pure n-hexane atmosphere, the Ti concentration was found to be below 1 at %, whereas in pure argon it reached 20 at %, as measured by Rutherford backscattering spectroscopy and elastic recoil detection analysis (RBS/ERDA). A high level of titanium oxidation is detected with TiO(2), substoichiometric titania, and titanium carbide, composing an inorganic phase of the composite films. In addition, high hydrogen content is detected in films rich with titanium. Ti-deficient and Ti-rich films proved equally good substrates for adhesion and growth of cultured human osteoblast-like MG 63 cells. In these cells, the population densities on days 1, 3, and 7 after seeding, spreading area on day 1, formation of talin-containing focal adhesion plaques as well as concentrations of talin and osteocalcin (per mg of protein) were comparable to the values obtained in cells on the reference cell culture materials, represented by microscopic glass coverslips or a polystyrene dish. An interesting finding was made when the Ti/ppCH films were seeded with calf pulmonary artery endothelial cells of the line CPAE. The cell population densities, the spreading area and also the concentration of von Willebrand factor, a marker of endothelial cell maturation, were significantly higher on Ti-rich than on Ti-deficient films. On Ti-rich films, these parameters were also higher or similar in comparison with the reference cell culture materials. Thus, both types of films could be used for coating bone implants, of which the Ti-rich film remains effective in enhancing the endothelialization of blood contacting artificial materials. PMID- 18384162 TI - Photoreversibility and photostability in films of octopus rhodopsin isolated from octopus photoreceptor membranes. AB - In this work, a new biomaterial resulting from the isolation of octopus rhodopsin (OR) starting from octopus photoreceptor membranes is presented. Mass spectroscopic characterization was employed in order to verify the presence of rhodopsin in the extract. Photoreversibility and photochromic properties were investigated using spectrophotometric measurements and pulsed light. Thin films of OR were realized using the gel-matrix entrapment method in polyvinyl alcohol solution. The results indicate that the photoreversibility and the photostability of the OR in gel-matrices are maintained. Several measurements were performed to test the stability of the resulting biomaterial in time and at room temperature. Preliminary tests demonstrate that the photoreversibility and the photostability are still found after few days from the biomaterial preparation and after the exposure for several hours at room temperature. PMID- 18384163 TI - Characterization of low-molecular-weight hyaluronic acid-based hydrogel and differential stem cell responses in the hydrogel microenvironments. AB - Hyaluronic acid is a natural glycosaminoglycan involved in biological processes. Low-molecular-weight hyaluronic acid (10 and 50 kDa)-based hydrogel was synthesized using derivatized hyaluronic acid. Hyaluronic acid was acrylated by two steps: (1) introduction of an amine group using adipic acid dihydrazide, and (2) acrylation by N-acryloxysuccinimide. Injectable hyaluronic acid-based hydrogel was prepared by using acrylated hyaluronic acid and poly(ethylene glycol) tetra-thiols via Michael-type addition reaction. Mechanical properties of the hydrogel were evaluated by varying the molecular weight of acrylated hyaluronic acid (10 and 50 kDa) and the weight percent of hydrogel. Hydrogel based on 50-kDa hyaluronic acid showed the shortest gelation time and the highest complex modulus. Next, human mesenchymal stem cells were cultured in cell adhesive RGD peptide-immobilized hydrogels together with bone morphogenic protein 2 (BMP-2). Cells cultured in the RGD/BMP-2-incorporated hydrogels showed proliferation rates higher than that of control or RGD-immobilized hydrogels. Real-time RT-PCR showed that the expression of osteoblast marker genes such as CBFalpha1 and alkaline phosphatase was increased in hyaluronic acid-based hydrogel, and the expression level was dependent on the molecular weight of hyaluronic acid, RGD peptide, and BMP-2. This study indicates that low-molecular weight hyaluronic acid-based hydrogel can be applied to tissue regeneration as differentiation guidance materials of stem cells. PMID- 18384164 TI - Rheological, microstructural, and in vitro characterization of hybrid chitosan polylactic acid/hydroxyapatite composites. AB - In this work, hybrid chitosan/hydroxyapatite composites material were developed and characterized. The polymer matrix was first dissolved in polylactic acid, and then hydroxyapatite (HA) was added as filler material. The effects of the added amounts of a crosslinking agent (genipin) and of the concentrations of lactic acid, and of the presence of HA powder on the evolution of rheological properties were evaluated. A significant decrease of gelation time with increasing amounts of crosslinking agent was observed, the effect being even more pronounced in the presence of HA. The chitosan matrix and the composites with a chitosan/HA weight ratio of 2/5 were characterized using microstructural analysis and in vitro tests. The formation of large pore sizes in the chitosan-based scaffolds was favored by low concentrations of lactic acid and genipin. The in vitro tests in synthetic body fluid revealed an extensive formation of an apatitic layer onto the surface of the chitosan/HA composite scaffolds crosslinked with genipin. PMID- 18384165 TI - A new generation of bio-derived ceramic materials for medical applications. AB - A new generation of bio-derived ceramics can be developed as a base material for medical implants. Specific plant species are used as templates on which innovative transformation processes can modify the chemical composition maintaining the original biostructure. Building on the outstanding mechanical properties of the starting lignocellulosic templates, it is possible to develop lightweight and high-strength scaffolds for bone substitution. In vitro and in vivo experiments demonstrate the excellent biocompatibility of this new silicon carbide material (bioSiC) and how it gets colonized by the hosting bone tissue because of its unique interconnected hierarchic porosity, which opens the door to new biomedical applications. PMID- 18384166 TI - Activated Src kinases interact with the N-methyl-D-aspartate receptor after neonatal brain ischemia. AB - OBJECTIVE: Neonatal stroke is associated with the N-methyl-D-aspartate receptor (NMDAR)-mediated excitotoxic brain injury. Src family kinases (SFKs) are considered to be the molecular hub for NMDAR regulation. We determined the relationship between SFKs activation and NMDAR tyrosine phosphorylation after neonatal hypoxia-ischemia (HI) and investigated the neuroprotective potential of a selective SFKs inhibitor, PP2 (4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo [3, 4-d] pyramidine), against neonatal brain ischemic injury. METHODS: The Rice Vannucci model was adapted for neonatal HI injury in postnatal day 7 CD1 mice. SFKs activity in the postsynaptic densities was measured by Western blot. NMDAR tyrosine phosphorylation and their association with SFKs were determined by coimmunoprecipitation. Brains from animals treated with PP2 or its inactive analog, PP3, were examined histologically with cresyl violet and iron stain to assess the degree of damage. RESULTS: Neonatal HI resulted in a rapid and transient increase in tyrosine phosphorylation of NMDAR subunits NR2A and NR2B. This upregulation correlated with the enhanced association of Fyn and Src with NR2A and NR2B. SFKs were activated in the postsynaptic densities after HI. Inhibition of SFKs with PP2 attenuated brain injury after neonatal HI, whereas PP3 did not protect the brain from the HI insult. INTERPRETATION: SFKs may play an important role in NMDAR-mediated excitotoxicity and downstream events leading to neuronal death after neonatal HI. Inhibition of SFKs may provide protection against neonatal stroke. Rather than blockade of NMDAR after HI in the developing brain, it may be safer and more beneficial to manipulate components of the NMDAR signaling complex at the postsynaptic density. PMID- 18384167 TI - Neonate hippocampal volumes: prematurity, perinatal predictors, and 2-year outcome. AB - OBJECTIVE: To compare preterm (PT) and full-term (FT) infant hippocampal volumes and to investigate the relations among PT hippocampal volume, perinatal risk factors, and neurodevelopmental outcome. METHODS: A total of 184 PT and 32 full term infants underwent magnetic resonance imaging at term equivalent age with manual segmentation of the hippocampi on coronal slices. Perinatal data were collected and 2-year neurodevelopment was evaluated with the Mental Development Index and Psychomotor Development Index on the Bayley Scales of Infant Development. RESULTS: PT and FT infant hippocampi did not significantly differ after controlling for head size, and percentage reductions in PT hippocampi (3.4%) were less than for cortical (7%) and deep nuclear gray matter (13%), and total brain tissue volume (4.7%). PT hippocampal volumes were significantly lower in infants with moderate-to-severe white matter injury (p < 0.001), exposure to postnatal steroids (right, p = 0.001; left, p = 0.008), and indomethacin treatment (right, p = 0.01; left, p = 0.03). PT infant hippocampal volumes correlated with the Mental (p < 0.001) and Psychomotor Development Indices (right, p = 0.001; left, p = 0.002) after correcting for head size and sex, but remained significant only for the Mental Development Index and left hippocampi (p = 0.04) after additionally adjusting for white matter injury and steroids. INTERPRETATION: Hippocampal volumes were reduced in PT infants exposed to several perinatal events but were preserved in PT infants without these exposures. Smaller PT hippocampal volumes were indirectly associated with delayed development at 2 years. PMID- 18384168 TI - Anti-MuSK patient antibodies disrupt the mouse neuromuscular junction. AB - OBJECTIVE: A subset of myasthenia gravis patients that are seronegative for anti acetylcholine receptor (anti-AChR) antibodies are instead seropositive for antibodies against the muscle-specific kinase (anti-MuSK-positive). Here, we test whether transfer of IgG from anti-MuSK-positive patients to mice confers impairment of the neuromuscular junction and muscle weakness. METHODS: IgG from anti-MuSK-positive myasthenia gravis patients or control IgG (seronegative for AChR and MuSK) was injected intraperitoneally (45 mg daily for 14 days) into 6 week-old female FVB/NJ and C57BL/6J mice. Changes at neuromuscular junctions in the tibialis anterior and diaphragm muscles were assessed by confocal fluorescent imaging of AChRs stained with fluorescent-alpha-bungarotoxin. Loss of function was assessed by electromyography. RESULTS: In experimental mice injected with anti-MuSK-positive patient IgG, postsynaptic AChR staining was reduced to as little as 22% of that seen in control mice. Experimental mice showed reduced apposition of the nerve terminal (labeled with antibodies against synaptophysin and neurofilament) and the postsynaptic AChR cluster (labeled with fluorescent alpha-bungarotoxin). Mice injected with IgG from two of three anti-MuSK-positive patients lost weight and developed muscle weakness associated with a decremental electromyographic trace on repetitive nerve stimulation. INTERPRETATION: IgG from anti-MuSK-positive patients can cause myasthenia gravis when injected into mice. This may be explained by a progressive reduction in the density of postsynaptic AChR combined with changes in the nerve terminal and its relation to the postsynaptic structure. PMID- 18384169 TI - Fabrication of burst pressure competent vascular grafts via electrospinning: effects of microstructure. AB - In this work, electrospun tubes of interest for vascular tissue engineering were fabricated and evaluated for burst pressure and suture retention strength (SRS) in the same context as tensile strength providing a direct, novel comparison. Tubes could be fabricated displaying average burst pressures up to 4000 mmHg- well above the standard of 2000 mmHg--and SRS values matching those of relevant natural tissues. Surprisingly, highly oriented fiber and maximal tensile properties are not absolutely necessary to attain clinically adequate burst pressures. The ability to resist bursting is clearly related to both initial solution solids loading and electrospinning deposition time. We make novel in situ observations of the relative microstructural characteristics of failure during bursting, and connect this to the conditions used to fabricate the graft. Processes typically thought to promote fiber alignment are, in fact, highly condition-dependent and do not always provide superior properties. In fact, electrospun structures displaying no discernable alignment could achieve burst pressures regarded clinically sufficient. The properties of individual electrospun fiber clearly do not fully dictate macroscale properties. Normal background levels of point bonding are enhanced by increased rotational speeds, and can have effects on properties more dominant than those of alignment. PMID- 18384170 TI - Interfaces in graded coatings on titanium-based implants. AB - Graded bilayered glass-ceramic composite coatings on Ti6Al4V substrates were fabricated using an enameling technique. The layers consisted of a mixture of glasses in the CaO-MgO-Na(2)O-K(2)O-P(2)O(5) system with different amounts of calcium phosphates (CPs). Optimum firing conditions have been determined for the fabrication of coatings having good adhesion to the metal, while avoiding deleterious reactions between the glass and the ceramic particles. The final coatings do not crack or delaminate. The use of high-silica layers (>60 wt % SiO(2)) in contact with the alloy promotes long-term stability of the coating; glass-metal adhesion is achieved through the formation of a nanostructured Ti(5)Si(3) layer. A surface layer containing a mixture of a low-silica glass ( approximately 53 wt % SiO(2)) and synthetic hydroxyapatite particles promotes the precipitation of new apatite during tests in vitro. The in vitro behavior of the coatings in simulated body fluid depends both on the composition of the glass matrix and the CP particles, and is strongly affected by the coating design and the firing conditions. PMID- 18384171 TI - Biodegradable fumarate-based drug-delivery systems for ophthalmic applications. AB - The function of a photocrosslinked poly(propylene fumarate) (PPF)/poly(N-vinyl pyrrolidone) (PVP) matrix for the sustained release of three ophthalmic model drugs, acetazolamide (AZ), dichlorphenamide (DP), and timolol maleate (TM), was investigated. The drugs differ in molecular weight and degree of dissociation in aqueous environments; both are parameters that significantly influence drug diffusivity. AZ, DP, and TM-loaded cylindrical rods (10 mm length, 0.6 mm diameter) were fabricated by photoinduced cross-copolymerization of PPF and N vinyl pyrrolidone (NVP) in molds. The released amounts of AZ, DP, TM, and NVP were determined by high-performance liquid chromatography (HPLC). The effects of drug properties and loading on the release kinetics were investigated. The in vitro release of AZ, DP, and TM was well sustained from the polymer matrices over a period of approximately 210, 270, and 250 days, respectively. The release kinetics correlated with the HPLC retention profiles of the different drugs. Following a small initial burst release (<10%), a dual modality release controlled by diffusion and bulk erosion was found for all drugs. Drug release rates of up to 4 microg/day were reached. Matrix drug loading generally affected the extent of the burst release, release kinetics, as well as the matrix water content and matrix degradation that were determined gravimetrically. Microcomputed tomography was used to image structural and dimensional changes of the devices. A preliminary rabbit implantation study revealed promising ocular biocompatibility of drug-free PPF/PVP matrices. All results indicate the potential of photocrosslinked PPF-based matrices as polymeric carriers for long term ophthalmic drug delivery. PMID- 18384172 TI - Effects of repetitive and short time strain in human bone marrow stromal cells. AB - For the long-term objective to engineer a functional bone construct, we applied unidirectional cyclic mechanical strain to human bone marrow stromal cells. Two strain regimes were applied comprising single and repetitive strain, respectively. For the single strain, we applied 15 and 60 min of mechanical load (1 Hz, 5% elongation). Increased proliferation and type I collagen levels were detected after 15- and 60-min load accompanied by increased type III collagen production after 60 min of strain. To study a possible amplification of effects, long time strain on 3 consecutive days, with every day 8 h strain duration, was applied in order to induce persistent and evident cellular reactions. Furthermore, an increased calcification was observed. The observed changes were not associated with changes in p38, extracellular signal regulated kinase or c jun N-terminal kinase activation (Western Blot). In conclusion, application of a single strain period of up to 60 min is not sufficient to induce persistent cellular reactions. Fifteen minutes seems to induce beneficial effects, whereas 60 min of strain induces collagen type III (as a sign of scar formation). Repetition of strain (3 x 8 h) induced enhanced differentiation without detrimental side effects. PMID- 18384173 TI - Poly(L-lactide)-b-poly(ethylene oxide) copolymers with different arms: hydrophilicity, biodegradable nanoparticles, in vitro degradation, and drug release behavior. AB - Biodegradable and amphiphilic poly(L-lactide)-b-poly(ethylene oxide) copolymers with different arms (PLLA-b-PEO having one, two, four, and six arms) were successfully synthesized via a two-step synthetic strategy. The hydrophilicity hydrophobicity balance of these copolymers was mainly controlled by both the arm number of copolymers (i.e., macromolecular architecture) and the poly(ethylene oxide) (PEO) composition. Biodegradable nanoparticles could be generated by direct injection of these PLLA-b-PEO copolymers solutions into distilled water, and their critical micelles concentrations decreased with the increasing arm number of copolymers. Moreover, both the hydrophilic PEO composition and the arm number of copolymers controlled the average size of PLLA-b-PEO nanoparticles, and the nanoparticles with adjustable sizes (20-85 nm) completely meet the size prerequisite (less than 100 nm) for targeted drug delivery. In vitro degradation of PLLA-b-PEO nanoparticles showed that the PLLA composition gradually increased over the degradation time, and the degree of crystallinity of PLLA block within copolymers increased simultaneously. Furthermore, the nimodipine drug loading efficiency of the PLLA-b-PEO copolymers was apparently higher than that of PLLA homopolymers. The drug-release experiments demonstrated that these biodegradable nanoparticles might be used for a short-time controlled release system. Consequently, this will provide a facile method not only to design new PLLA-based biomaterials from both the macromolecular architecture and the hydrophilicity hydrophobicity balance, but also to fabricate biodegradable nanoparticles with adjustable sizes for drug delivery. PMID- 18384174 TI - In situ observation of surface oxide layers on medical grade Ni-Ti alloy during straining. AB - Medical grade Ni-Ti alloys with shape memory or pseudo-elastic behavior exhibit good biocompatibility because of an electrochemically passive oxide layer on the surface. In this work, the mechanical stability of surface oxide layers is investigated during reversible pseudo-elastic deformation of commonly applied medical grade Ni-Ti wires. Surface oxide layers with varying thickness were generated by varying annealing times under air atmosphere. The thicknesses of the surface oxide layers were determined by means of Rutherford backscattering spectrometry. In situ scanning electron microscopy investigations reveal a damage mechanism, which is assumed to have a significant influence on the biocompatibility of the material. The conditions that lead to the appearance of cracks in the surface oxide layer or to the flaking of surface oxide layer particles are identified. The influence of the thickness of the surface oxide layer on the damage mode is characterized. The possible impact of the damaged surface oxide layer on the material's biocompatibility and the potentials to reduce or avoid the damage are discussed. PMID- 18384176 TI - Clinical application of MRI in ophthalmology. AB - MRI has long been applied to clinical medical and neurological cases for the structural assessment of tissues as well as their physiological and functional needs and processes. These uses are at a variety of developmental stages in ophthalmology, from common use of clinical structural assessment for neuro ophthalmology and evaluation of space-occupying lesions to the beginning stages of experimentally measuring functional activation of specific layers within the retina and measurement of physiological oxygen responses. New MRI methodologies, such as the use of orbital coils and Gd-DTPA image enhancement, have been researched, developed, and validated in the eye, opening new possibilities for this technology to enter the clinic. This review aims to summarize the clinical ophthalmological uses of MRI, focusing on the current use of the technology and future applications. PMID- 18384177 TI - Experimental comparison of four FAIR arterial spin labeling techniques for quantification of mouse cerebral blood flow at 4.7 T. AB - Pulsed arterial spin labeling (ASL) is an attractive and robust method for quantification of rodent cerebral blood flow (CBF) in particular, although there is a need for sensitivity optimization. Look-Locker flow-sensitive alternating inversion recovery (FAIR) echo planar imaging (EPI) (LLFAIREPI) was expected to be a likely candidate for assessing sensitivity, although it has not yet been applied to rodents. In this study, the performance of two FAIR techniques and two Look-Locker FAIR techniques were compared in mouse brain at 4.7 T. FAIR-EPI (single inversion time, FAIREPI-1TI), FAIR-EPI (eight inversion times, FAIREPI 8TI), LLFAIREPI and Look-Locker FAIR gradient echo (LLFAIRGE) sequences were implemented with equal spatial resolution and equal FAIR preparation modules. Measurements were carried out sequentially on the brain in 10 healthy mice, and quantitative CBF maps were obtained after different acquisition times up to 23 min. All methods gave similar group variability in CBF. Especially at shorter acquisition times, LLFAIREPI gave lower relative variations in CBF within selected brain regions than the other techniques at the same acquisition time. The Look-Locker techniques, however, overestimated CBF compared with classical FAIR-EPI, which was attributed to bulk flow in arterioles and T(2) effects. The image quality with LLFAIREPI was less reproducible within the group. Both FAIREPI 1TI and LLFAIREPI appear to be good candidates for serial rapid measurement of CBF, but LLFAIREPI has the additional advantage that apparent T(1) can be measured simultaneously with CBF. PMID- 18384178 TI - Metabolic viability assessment of cystic echinococcosis using high-field 1H MRS of cyst contents. AB - Cystic echinococcosis is a worldwide disease caused by larval stages of the parasite Echinococcus granulosus (canine tapeworm). In clinical practice, staging of cyst development by ultrasonography (US) has allowed treatment options to be tailored to individual patient needs. However, the empirical correlation between cyst morphology and parasite viability is not always dependable and has, until now, required confirmation by invasive assessment of cyst content by light microscopy (LM), for example. Alternatively, high-field 1H MRS may be used to examine cyst fluid ex vivo and prepare detailed quantitative metabolite profiles, enabling a multivariate metabolomics approach to cyst staging. One-dimensional and two-dimensional 1H and 1H/13C MRS at 600 MHz (14.1 T) was used to analyze 50 cyst aspirates of various US and LM classes. MR parameters and concentrations relative to internal valine were determined for 44 metabolites and four substance classes. The high concentrations of succinate, fumarate, malate, acetate, alanine, and lactate found in earlier studies of viable cysts were confirmed, and additional metabolites such as myo-inositol, sorbitol, 1,5-anhydro-D-glucitol, betaine, and 2-hydroxyisovalerate were identified. Data analysis and cyst classification were performed using univariate (succinate), bivariate (succinate vs fumarate), and multivariate partial least squares discriminant analysis (PSL DA) methods (with up to 48 metabolite variables). Metabolic classification of 23 viable and 18 nonviable cysts on the basis of succinate alone agreed with LM results. However, for seven samples, LM and MRS gave opposing results. Reclassification of these samples and two unclassified samples by PLS-DA prediction techniques led to a set of 50 samples that could be completely separated into viable and nonviable MRS classes with no overlap, using as few as nine variables: succinate, formate, malate, 2-hydroxyisovalerate, acetate, total protein content, 1,5-anhydro-D-glucitol, alanine, and betaine. Thus, future noninvasive in vivo applications of MRS would appear promising. PMID- 18384180 TI - Quantification of cerebral edema on diffusion tensor imaging in acute-on-chronic liver failure. AB - Cerebral edema is a major complication of acute liver failure but may also be seen in other forms of liver failure such as acute-on-chronic liver failure (ACLF) and chronic liver failure (CLF). ACLF develops in patients with previously well-compensated chronic liver disease following acute hepatitis A or E superimposed on underlying liver cirrhosis. The aim of this study was to detect the occurrence, and determine the nature, of cerebral edema in patients with the defined subset of ACLF using diffusion tensor imaging (DTI) metrics. Twenty-three patients with ACLF were studied and compared with 15 healthy controls and 15 patients with CLF. DTI metrics including fractional anisotropy (FA), mean diffusivity (MD), linear anisotropy (CL), planar anisotropy (CP), and spherical isotropy (CS) were calculated by selecting regions of interest in the white matter and deep grey matter of the brain. Significantly decreased FA and increased CS were observed in the anterior limb (ALIC) and posterior limb (PLIC) of the internal capsule and frontal white matter (P<0.05) in patients with different grades (1-4) of ACLF when compared with healthy controls. No significant changes in MD and CP were seen in any brain region. However, significantly decreased CL was observed in the PLIC, caudate nuclei and putamen. In patients with CLF, significantly decreased FA with increased CS in the ALIC and PLIC along with significantly increased MD in the ALIC and caudate nuclei were observed. The presence of significantly decreased FA and CL and increased CS along with no significant change in MD and CP suggests the presence of both intracellular and extracellular components of cerebral edema in patients with ACLF. PMID- 18384179 TI - Numerical field calculations considering the human subject for engineering and safety assurance in MRI. AB - Numerical calculations of static, switched, and radiofrequency (RF) electromagnetic (EM) fields considering the geometry and EM properties of the human body are used increasingly in MRI to explain observed phenomena, explore the limitations of various approaches, engineer improved techniques and technology, and assure safety. As the static field strengths and RF field frequencies in MRI have increased in recent years, the value of these methods has become more pronounced and their use has become more widespread. With the recent growth in parallel reception techniques and the advent of transmit RF arrays, the utility of these calculations will become only more critical to continued progress of MRI. Proper relation of field calculation results to the MRI experiment can require significant understanding of MRI physics, EM field principles, MRI coil hardware, and EM field safety. Here some fundamental principles are reviewed and current approaches and applications are catalogued to aid the reader in finding resources valuable in beginning field calculations for their own applications in MR, with an eye to the current needs and future utility of numerical field calculations in MRI. PMID- 18384182 TI - Longitudinal study of the assessment by MRI and diffusion-weighted imaging of tumor response in patients with locally advanced breast cancer undergoing neoadjuvant chemotherapy. AB - Measurements of tumor apparent diffusion coefficient (ADC), volume and diameter in assessing the response of patients with locally advanced breast cancer (LABC) (n = 56) undergoing neoadjuvant chemotherapy (NACT) at four time periods (before treatment and after three cycles of NACT) were carried out at 1.5 T using diffusion-weighted imaging (DWI) and MRI. Ten benign tumors and 15 controls were also investigated. The MR tumor response was compared with the clinical response. Mean ADC before treatment of malignant breast tissue was significantly lower than that of controls, disease-free contralateral tissue of the patients, and benign lesions, and gradually increased during the course of NACT. Analysis of the percentage change in ADC, volume and diameter after each cycle of NACT between clinical responders and non-responders showed that the change in ADC after the first cycle was statistically significant compared with volume and diameter, indicating its potential in assessing early response. After the third cycle, the sensitivity for differentiating responders from non-responders was 89% for volume and diameter and 68% for ADC, and the respective specificities were 50%, 70% and 100%. A sensitivity of 84% (specificity of 60% with an accuracy of 76%) was achieved when all three variables were taken together to predict the response. A cut-off value of ADC was also calculated using receiver operator characteristics analysis to discriminate between normal, benign and malignant breast tissue. Similarly, a cut-off value for ADC, volume and diameter was obtained after the second and third cycles of NACT to predict tumor response. The results show that ADC is more useful for predicting early tumor response to NACT than morphological variables, suggesting its potential in effective treatment management. PMID- 18384181 TI - In vivo MRS markers of response to CHOP chemotherapy in the WSU-DLCL2 human diffuse large B-cell lymphoma xenograft. AB - To identify 1H-MRS molecular biomarkers of early clinical therapeutic response in non-Hodgkin's lymphoma, an in vivo longitudinal study was performed on human non Hodgkin's diffuse large B-cell lymphoma xenografts (WSU-DLCL2) grown in the flanks of female SCID mice. 31P-MRS measurements, which have been demonstrated to be prognostic clinical indices of response (Arias-Mendoza et al. Acad. Radiol. 2004; 11: 368-376) but which provide lower spatial resolution, were included for comparison. The animals received CHOP (cyclophosphamide, hydroxydoxorubicin, oncovin and prednisone) chemotherapy for three 1-week cycles, resulting in stable disease based on tumor volume. Localization of total choline and phosphorus metabolites in vivo was achieved with stimulated echo acquisition mode and image selected in vivo spectroscopy sequences, respectively. Significant decreases in lactate were detected by the selective multiple quantum coherence spectral editing technique after the first cycle of CHOP, whereas total choline and the phosphomonoester/nucleoside triphosphate ratio did not change until the third cycle. Ex vivo extract MRS of tumors corroborated the in vivo results. Histological staining with antibodies to Ki67 revealed a decrease in proliferation rate in CHOP-treated tumors that coincided with the decrease in lactate. This study demonstrates the utility of lactate as an early proliferation sensitive indicator of therapeutic response in a mouse model of non-Hodgkin's lymphoma and serves as a basis for future clinical implementation of these methods. PMID- 18384183 TI - Small area random effects models for capture/recapture methods with applications to estimating coverage error in the U.S. Decennial Census. AB - The synthetic estimation approach currently in use for estimating net coverage error in the U.S. Census is evaluated using random effects models. The synthetic estimates from the 2000 Accuracy and Coverage Evaluation (ACE) Revision II are evaluated in two parts. First, a model is used, which produces the synthetic estimate components and, second, the model is enlarged to include random effects at the small area level. Retaining all the fixed effects that characterize the synthetic model produces an extremely large, saturated random effects model. Hence, we selectively reduce the random effects model with an aim towards keeping all fixed effects in order to fairly evaluate the synthetic model. A super population model is used for the bivariate outcome of erroneous enumeration rate and census omission rate. Both these outcomes were previously estimated using the current synthetic estimation approach. A major hurdle in this project was the development of defensible input data for the small areas due to the large number of effects in the synthetic model, which render simple design-based estimates for small areas crossed with post-strata, mostly, unusable. For this initial approach, the small areas were the 540 local census offices. Bayesian methods are employed to evaluate these models. The advantage of this model is that it can evaluate a key assumption about the homogeneity of rates within a post-stratum and if the assumption holds, then this model reduces to the current synthetic model. PMID- 18384185 TI - A robust P-value for treatment effect in meta-analysis with publication bias. AB - Publication bias is a major and intractable problem in meta-analysis. There have been several attempts in the literature to adapt methods to allow for such bias, but these are only possible if we are prepared to make strong assumptions about the underlying selection mechanism. We discuss the assumption that the probability that a paper is published may depend in some unspecified way on the P value being claimed by that study. We suggest a new robust P-value for the overall treatment effect, which turns out to be closely related to the correlation of the associated radial plot. Properties of the method are discussed and illustrated in two examples. PMID- 18384186 TI - The Sertindole Cohort Prospective (SCoP) study: rationale, design and methodology. AB - PURPOSE: To describe the rationale and methodology of the Sertindole Cohort Prospective (SCoP) study. METHODS: The SCoP study was a prospective, randomized, partially blinded, active-controlled, multinational trial. It was designed to assess the safety of the antipsychotic sertindole in the treatment of schizophrenia under normal conditions of use. Risperidone, a widely used antipsychotic, not associated with major safety concerns, served as comparator drug. Inclusion criteria were deliberately broad in order to ensure high external validity, but patients had to be eligible for treatment with both drugs. The first primary endpoint was all-cause mortality, a measure highly resistant to bias, and the second was hospitalization with arrhythmia. Secondary endpoints comprised cause-specific fatal events, hospitalizations, suicide attempts and treatment duration. An Independent Safety Committee (ISC) classified the events using blinded data and provided advice to an Independent Management Committee (IMC) that was overseeing the trial. It was calculated that 3800 person years of exposure were needed in each treatment arm to obtain a power of 80%. This makes the SCoP study one of the largest post-authorization trials ever conducted in schizophrenia research. RESULTS: This report describes the design of the study, its rationale and the methods used. CONCLUSIONS: Naturalistic estimates of drug safety constitute essential information when prescribing antipsychotic medication. In the SCoP study, data were collected prospectively in a randomized, controlled manner and under normal conditions of use. Such a design ensured the high quality of data needed to adequately evaluate the 'real-world' safety of sertindole treatment. PMID- 18384187 TI - Investigating the effects of ties on measures of concordance. AB - The concordance between predicted and observed outcomes, referred to as the C index or C statistic, is frequently used to quantify the discriminatory ability of a prognostic model. It also commonly serves as a basis for distinguishing predictive strength between different models. Two alternative formulations of the C index are widely used, one completely excluding ties from the computation (Ctied,out) and the other including ties (Ctied,in). However, there has been little research concerning the effects of ties on these two measures. In this paper we characterize changes in the performance of Ctied,in and Ctied,out for progressively less coarse (or more coarse) partitions of the data. Our theoretical and simulation results show that both measures can be heavily dependent on the number of tied pairs and their results can be substantially divergent. We examine potential ambiguities that can occur when the two measures diverge. In the presence of a substantial proportion of tied pairs, we recommend that both C indices be computed as lower and upper bounds, and that, when feasible, the C indices should be computed with respect to partitions corresponding to the same percentiles for the models being compared. When it is desired to estimate the overall concordance, the average value of the two C indices might also be considered. We use the data from the Hemodialysis Clinical Trial to illustrate our evaluation and methods. PMID- 18384188 TI - Antiinflammatory activities of flavonoids and a triterpene caffeate isolated from Bauhinia variegata. AB - In the continuing search for novel antiinflammatory agents, six flavonoids, namely kaempferol (1), ombuin (2), kaempferol 7,4'-dimethyl ether 3-O-beta-D glucopyranoside (3), kaempferol 3-O-beta-D-glucopyranoside (4), isorhamnetin 3-O beta-D-glucopyranoside (5) and hesperidin (6), together with one triterpene caffeate, 3beta-trans-(3,4-dihydroxycinnamoyloxy)olean-12-en-28-oic acid (7) were isolated from the non-woody aerial parts of Bauhinia variegata. Compounds 1-7 were evaluated as inhibitors of some macrophage functions involved in the inflammatory process. These seven compounds significantly and dose dependently inhibited lipopolysaccharide (LPS) and interferon (IFN)-gamma induced nitric oxide (NO), and cytokines [tumor necrosis factor (TNF)-alpha and interleukin (IL) 12]. The concentration causing a 50% inhibition (IC50) of NO, TNF-alpha and IL-12 production by compounds 1, 2 and 7 was approximately 30, 50 and 10 microM, respectively, while at 50, 200 and 40 microM compounds 3, 4, and 5, 6 showed 15 30% inhibition, respectively. On the other hand, compounds 3 and 7 showed no inhibitory effect, while compounds 1, 4-6 reduced by around 10-30% the synthesis of NO by macrophages, when inducible NO synthase was already expressed with LPS/IFN-gamma for 24 h. These experimental findings lend pharmacological support to the suggested folkloric uses of the plant B. variegata in the management of inflammatory conditions. PMID- 18384189 TI - Comments on 'The performance of different propensity score methods for estimating marginal odds ratios'. PMID- 18384191 TI - A systematic review on the Rosa canina effect and efficacy profiles. AB - Rose hip, rose hip and seed and rose hip seed, all were negatively monographed by the German Commission E due to insufficient evidence of effects and effectiveness. Therefore a comprehensive review of the literature was conducted to summarize the pharmacological and clinical effects of Rosa canina L. to reevaluate its usefulness in traditional medicine. For various preparations of rose hip and rose hip and seed, antioxidative and antiinflammatory effects have been demonstrated. Lipophilic constituents are involved in those mechanisms of action. The proprietary rose hip and seed powder Litozin has been employed successfully in a number of exploratory studies in patients suffering from osteoarthritis, rheumatoid arthritis and low back pain. However, the sizes of the clinical effects for the different indications need to be determined to assure clinical significance. There is also a rationale behind the use of Litozin as part of a hypocaloric diet based on the rose hip probiotic, stool regulating and smooth muscle-relaxing actions, as well as the rose hip seed lipid-lowering, antiobese and antiulcerogenic effects. Further research is needed to clarify the importance of the reported promising experimental effects in clinical use and to characterize the optimum rose hip seed oil preparation for topical use in the treatment of skin diseases. PMID- 18384190 TI - Aqueous extract of rhubarb stabilizes vulnerable atherosclerotic plaques due to depression of inflammation and lipid accumulation. AB - The study evaluated the effect of the traditional Chinese medicine rhubarb on the stability of atherosclerotic plaque. Atherosclerotic lesions were induced in rabbits through balloon injury with a high-cholesterol diet and then were divided into a control group, a rhubarb group and a simvastatin group. At week 24 recombinant-p53 adenoviruses were locally delivered to the atherosclerotic plaques. At week 26 plaque rupture was triggered by the intra-arterial Chinese Russell's viper venom and histamine. Serological, ultrasonographic, pathologic, immunohistochemical and gene expression studies were performed. The results showed that the incidence of plaque rupture in the rhubarb group and the simvastatin group was significantly lower than that in the control group (42.86% and 35.71% versus 80.00%, both p < 0.05). Serum TC, LDL-C (p < 0.05-0.01), IMT (both p < 0.01), PA (both p < 0.01), PB (%) (both p < 0.01) and the mRNA and protein expressions of TLR2, TLR4 and NF-kappaB (p < 0.05, 0.01, respectively) in the rhubarb group and the simvastatin group were significantly lower than those in the control group. In contrast, AIIc% (both p < 0.05) in the two treatment groups were significantly higher than those in the control group. These results suggest that rhubarb has antiatherosclerotic and plaque-stabilizing properties due to antiinflammation and lipid-lowering effects. PMID- 18384192 TI - Herbal and food folk medicines of the Russlanddeutschen living in Kunzelsau/Talacker, South-Western Germany. AB - An urban ethnobotanical study was carried out among a community of Russlanddeutschen (Germans from Russia) who in recent years have moved from Russia and Central Asia to Kunzelsau, a small town located in Wurttemberg, in South-Western Germany. Thirty-six in-depth interviews were conducted with the women in this community, and 62 homemade medicinal preparations derived from 46 botanical species were recorded. As well as common medicinal plant uses that are well known in modern evidence-based German and Western European phytotherapy, we were able to record traces of the community's Russian and Central Asian (Turkic) heritage through the very popular use of sorrel as a depurative or for preventing and treating colds and flu; the use of dill as a digestive; watermelon as a diuretic; birch to relieve rheumatism and arthritis; buckwheat as a tonic; rye based fermented beverages as a stimulant and as a depurative, diverse berries to prevent colds and flu; coriander as a digestive, and other medicinal foods. Traces of archaic German preparations were also recorded, which were probably Swabian in origin. Nearly half of the overall quoted items represented folk functional foods. The researchers believe that the findings in this study could stimulate public health policies aimed at improving both the phyto pharmacovigilance of lesser-known herbal drugs, and the health and well-being of migrants by promoting a better understanding of emic health beliefs and newcomers' healing strategies. PMID- 18384193 TI - The tripole linear ion trap with highly efficient orthogonal ion ejection designed by computer simulations. AB - An ion guide, consisting of three rods carrying three alternating current (AC) voltages symmetrically delayed, called a tripole, was used as a linear ion trap (LIT) and studied by computer simulations. Radial containment of ions was also demonstrated with the pseudopotential which was calculated by approximating the tripole electric potential to the multipoles expansion. This work found a new analyte concentrator, which performs effective ion ejection, and is suitable for use with time-of-flight mass spectrometry. The efficiency of the overall process from the trapping until the ejection was higher than 90%, although some degree of ion spatial and kinetic energy spread which can be corrected with a reflectron was obtained. The reason for the ejection of this tripole linear ion trap (tLIT) lies in the high space available between the rods. The ejection is optimized with the application of focusing voltages, especially suitable for a tripole symmetry (one electrode has a pulse offset voltage and the other two have a fraction of that pulse). The beam is finally well parallelized with a rectangular Einzel lens. PMID- 18384194 TI - Hydrophilic interaction chromatography/electrospray mass spectrometry analysis of carbohydrate-related metabolites from Arabidopsis thaliana leaf tissue. AB - This work describes the development and application of an on-line liquid chromatography/mass spectrometry (LC/MS) method using hydrophilic interaction chromatography (HILIC) coupled to negative ion mode electrospray ionisation ion trap mass spectrometry (ESI-MS) for the analysis of highly polar carbohydrate related metabolites commonly found in plants, ranging from reducing and non reducing sugars and sugar alcohols to sugar phosphates. Using this method, separation and detection of a mixture of eight authentic standard compounds containing glucose (Glc), sucrose (Suc), raffinose, verbascose, mannitol, maltitol, glucose-6-phosphate (Glc6P) and trehalose-6-phosphate (Tre6P) were achieved in less than 15 min. The method is rapid, robust, selective, and sensitive, with limits of detection (LODs) ranging from 0.2 microM obtained for neutral sugars, to 1.0 microM obtained for sugar alcohols, and 2.0 microM obtained for negatively charged sugar phosphates. We have studied the negative ion collision-induced dissociation (CID) fragmentation behaviour of the non reducing raffinose family oligosaccharides (RFOs) raffinose, stachyose, and verbascose. Mainly Bi and Ci glycosidic and Ai cross-ring structurally informative cleavages are observed. We have applied this HILIC/ESI-MS method for the analysis of Arabidopsis thaliana wild-type Columbia-0 (Col-0) and its starchless phosphoglucomutase mutant (pgm1) leaf extracts. The method was used to quantify Glc, Suc, raffinose, and Glc6P in A. thaliana extracts. Data obtained using this HILIC/ESI-MS method were compared with those obtained using a comparable porous graphitic carbon-based LC/ESI-MS method. PMID- 18384195 TI - Rapid screening for cyclo-dopa and diketopiperazine alkaloids in crude extracts of Portulaca oleracea L. using liquid chromatography/tandem mass spectrometry. AB - A simple and rapid qualitative liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated for screening cyclo-dopa and diketopiperazine alkaloids in crude extracts of Portulaca oleracea L. at sub-ppm levels. An electrospray ionization orbitrap mass spectrometer, which provides accurate full scan MS and MS/MS data, was used in this study. After simple extraction with ethanol and purification by AB-8 resin, the extracts were subjected to LC/MS/MS analysis. A high mass tolerance (10 ppm) was used in the initial screening to filter the full scan MS data. The cyclo-dopa and diketopiperazine alkaloid standards gave limits of detection (LODs) at or below 5 ng/mL. The results also indicated that the method had an acceptable precision for day-to-day use in the identification of compounds. The alkaloids could be identified based on their MS/MS data, elemental compositions, and retention behavior. This system was used to assay trace amounts of cyclo-dopa and diketopiperazine alkaloids in crude extracts of Portulaca oleracea L., leading to the identification of 5/2 confirmed/unconfirmed cyclo-dopa and 7/6 confirmed/unconfirmed diketopiperazine alkaloids, respectively. The screening method considerably reduces the time and cost involved in the identification of cyclo-dopa and diketopiperazine alkaloids in Portulaca oleracea L., as well as being a simple and convenient approach to the identification of other structural families of natural products. PMID- 18384196 TI - Halogen-substituted phenylalanines as enantioselective selectors for enantioselective discrimination of amino acids: effect of halogen. AB - Halogen-substituted phenylalanines with a halogen X (X = F, Cl, Br or I) in the para position in the aromatic ring of L-phenylalanine are used as enantioselective selectors to explore the effect of the halogen substituent on the enantioselective discrimination of amino acids. Enantioselective discrimination is achieved by investigating the collision-induced dissociation spectra of the trimeric complex ion, [CuII(ref)2(A)-H]+, generated by electrospraying a solution of a mixture of D- or L-analyte amino acid (A), enantioselective reference ligand (ref) and CuCl2. The relative abundances of fragment ions resulting from the competitive loss of reference and analyte amino acids are considered for measuring the degree of enantioselective discrimination by applying the kinetic method. The enantioselectivity of the p-halogenated derivatives of L-Phe increases from fluorine to iodine for the studied amino acids (except for acidic amino acids). The validity of the present method has also been checked by cross enantioselective experiments using p-iodo-D phenylalanine as the reference in place of p-iodo-L-phenylalanine. The enantioselectivity of fluoro-substituted L-phenylalanine is less than that obtained with L-phenylalanine. The high inductive effect of the fluorine atom decreases the strength of the pi-pi stacking interaction. The presence of halogen affects the enantioselectivity by inductive and steric effects. PMID- 18384197 TI - Use of multiply charged atomic ions for isotope ratio mass spectrometry. AB - We have investigated the use of multiply charged atomic ions for the measurement of isotopic ratios of gaseous and vapour samples. We use a mass spectrometer system incorporating an electron cyclotron resonance (ECR) ion source for this purpose. In the cases of carbon, nitrogen and oxygen, the selection of the 2+ atomic species is found to be the most effective for obtaining reliable isotopic ratios. Using samples of carbon dioxide, nitrogen, air and water vapour, we have demonstrated the determination of the isotopic ratios 13C/12C, 15N/14N, 17 O/16 O and 18 O/16 O. For oxygen, this technique offers an alternative to the equilibration or purification methods normally required to obtain isotopic ratios of water or other oxygen-containing samples. In particular, 17 O/16 O can be measured directly without isobaric interference from OH+. With typical ionization efficiencies of greater than 10%, ECR ion sources have the potential to enable measurements on very small samples. In addition to those evaluated in the present work, there is scope for application of this method to other sample types, to a variety of sampling methods, and to other elements. PMID- 18384198 TI - Effects of the brown seaweed Undaria pinnatifida on erythematous inflammation assessed using digital photo analysis. AB - The brown seaweed Undaria pinnatifida (Harvey) Suringar produced potent inhibition of erythematous inflammation assessed using digital photo analysis. The analysis technique was validated by laser speckle flowgraphy and blood vessel contraction. The methanol extract suppressed erythema by 50% when applied within 1 h before or 15 min after application of phorbol myristate acetate. Erythema reduction to half-maximal values took 12 h with the extract, compared with 25 h with the vehicle. The blade part of the thallus showed the highest activity, while the northern type of U. pinnatifida had slightly higher activity than the southern type. The active constituents were stearidonic acid and eicosapentaenoic acid. These findings reinforce the claims of the health care industry and indigenous medicine that U. pinnatifida can be used as a health food and remedy for inflammation-related symptoms. PMID- 18384199 TI - Fermented papaya preparation as redox regulator in blood cells of beta thalassemic mice and patients. AB - Many aspects of the pathology in beta-hemoglobinopathies (beta-thalassemia and sickle cell anemia) are mediated by oxidative stress. Fermented papaya preparation (FPP) was tested for its antioxidant effects: the scavenging effect was determined spectrofluorometrically in a cell-free system using 2'-7' dichlorofluorescin-diacetate (DCF). Both spontaneous and H(2)O(2)-induced DCF oxidations were decreased by FPP in a dose-dependent fashion. Using flow cytometry, it was shown that in vitro treatment of blood cells from beta thalassemic patients with FPP increased the glutathione content of red blood cells (RBC), platelets and polymorphonuclear (PMN) leukocytes, and reduced their reactive oxygen species, membrane lipid peroxidation and externalization of phosphatidylserine. These effects result in (a) reduced thalassemic RBC sensitivity to hemolysis and phagocytosis by macrophages; (b) improved PMN ability to generate oxidative burst - an intracellular mechanism of bacteriolysis, and (c) reduced platelet tendency to undergo activation, as reflected by fewer platelets carrying external phosphatidylserine. Oral administration of FPP to beta-thalassemic mice (50 mg/mouse/day for 3 months) and to patients (3 g x 3 times/day for 3 months), reduced all the above mentioned parameters of oxidative stress (p < 0.001 in mice and p < 0.005 in patients). These results suggest that FPP, as a potent antioxidant, might alleviate symptoms associated with oxidative stress in severe forms of thalassemia. PMID- 18384200 TI - Disassembly of noncovalent amphiphilic polymers with proteins and utility in pattern sensing. AB - A simple strategy for pattern recognition of proteins through micellar disassembly is introduced. Five different noncovalently assembled receptors have been generated, and the disassembly was studied by monitoring the encapsulated dye release in response to five different proteins. The disassembly induced fluorescence change of the guest molecule produces protein-specific patterns. PMID- 18384201 TI - Fabrication of heterogeneous binary arrays of nanoparticles via colloidal lithography. AB - Heterogeneous binary arrays of metallic nanoparticles have been constructed by consecutively depositing gold and silver into monolayers of hexagonally close packed latex spheres at the incidence angles of 15 and -15 degrees, followed by removal of the colloidal masks. The present approach is independent of the chemical nature of both colloidal masks and deposition materials. The pattern feature of the resulting binary nanoparticle arrays is dependent on the colloidal mask registry. PMID- 18384202 TI - Development of a high-affinity anti-domoic acid sheep scFv and its use in detection of the toxin in shellfish. AB - The potential of immunoassays as high-throughput screening tools for the detection of harmful substances in foods will only be realized when convenient methods are available for production of the high affinity antibodies needed for sensitive assay development. Recombinant antibodies offer advantages over traditional monoclonal antibodies in terms of ease of production, much greater antibody repertoire for selection, and versatility. We describe here the development of recombinant antibodies against the common shellfish toxin, domoic acid (DA), utilizing the sheep immunoglobulin system as an effective method for generating high affinity anti-hapten recombinant antibody fragments. A single chain antibody fragment (scFv) library was generated from a sheep immunized with DA-bovine serum albumin conjugate, and anti-DA scFvs were isolated by phage display. Three selected scFvs gave I50s of 2.6 to 58 ng/mL (8.3-186 nM) in competitive enzyme-linked immunosorbent assay (ELISA). Assay optimization with one of these scFvs gave a very reproducible standard curve with a range of 0.3 to 5.6 ng/mL (1.0 to 17.9 nM), a mean limit of quantification (LOQ, defined as the I20) of 0.5 ng/mL (1.6 nM), and a mean I50 of 1.2 ng/mL (3.9 nM). When the assay was used for the analysis of crude methanolic extracts of scallop tissues, results obtained correlated well with standard HPLC assay results (R2, 0.90, n = 40; R2, 0.81, n = 34), although ELISA results were lower than HPLC results. Adjusting the cutoff point for DA concentration accordingly from the regulatory 20 mg/kg, the potential of the sheep scFv-based ELISA for use as a screening assay for DA in shellfish extracts was demonstrated. PMID- 18384203 TI - Capillary electrophoresis-fourier transform ion cyclotron resonance mass spectrometry for the identification of cationic metabolites via a pH-mediated stacking-transient isotachophoretic method. AB - Capillary electrophoresis-mass spectrometry (CE-MS) is still widely regarded as an emerging tool in the field of metabolomics and metabolite profiling. A major reason for this is a reported lack of sensitivity of CE-MS when compared to gas chromatography-mass spectrometry GC/MS and liquid chromatography-mass spectrometry. The problems caused by the lack of sensitivity are exacerbated when CE is coupled to Fourier transform ion cyclotron resonance mass spectrometry (FT ICR MS), due to the relatively low data acquisition rate of FT-ICR MS. Here, we demonstrate the use of an online CE sample preconcentration method that uses a combination of pH-mediated stacking and transient isotachophoresis, coupled with FT-ICR MS to improve the overall detection of cationic metabolites in the bacterium Desulfovibrio vulgaris Hildenborough. This method showed a significant increase in signal-to-noise ratio when compared to CE normal sample stacking, while providing good separation efficiency, reproducibility, and linearity. Detection limits for selected amino acids were between 0.1 and 2 microM. Furthermore, FT-ICR MS detection consistently demonstrated good mass resolution and sub-ppm mass accuracy. PMID- 18384204 TI - A synoptic report for bone marrow aspiration and core biopsies. PMID- 18384205 TI - Introduction to selected reviews based on the Massachusetts General Hospital, Harvard Medical School postgraduate course: "Current concepts in surgical pathology". PMID- 18384206 TI - Shirt-sleeve or scanning magnification as an aid to the diagnosis of commonly encountered medical lung diseases. AB - Shirt-sleeve magnification (holding a slide over a white sleeve) and low-power magnification serve as useful adjuncts in the general categorization of noninfectious medical lung disease. This article divides medical lung disease into chronic and acute, where the temporality is determined first by clinical circumstances and then confirmed by histopathology. The low-power patterns of various lung diseases overlap, sometimes greatly. Nevertheless, classic examples of chronic disease can be sorted as linear, lobular filling, nodular dispersed, nodular lymphangitic, or cystic patterns at shirt-sleeve or low-power magnification. Classic examples of acute disease generally produce a solidifying pattern at shirt-sleeve or low-power magnification, which can be followed by a determination as to whether alveolar filling is principally fibrotic or principally fluid or cells at higher magnification. This article gives a simple system for the categorization of medical lung disease by this approach, with an emphasis on the most common diseases to be encountered in a general surgical pathology practice. In our experience, this system also proves useful in arriving at some therapeutic decisions. PMID- 18384207 TI - Testicular tumors--some new and a few perennial problems. AB - The histopathology of testicular tumors is presented, emphasizing new, unusual, or underemphasized aspects. Within the category of seminoma of the usual type, the recent literature has drawn attention to the presence in occasional tumors of solid or hollow tubules or spaces of varying sizes and shape that may result in cribriform or microcystic patterns, causing potential confusion with other neoplasms, most notably Sertoli cell tumor or yolk sac tumor. Although regions of typical neoplasia and awareness of this phenomenon usually will be diagnostic, immunohistochemistry may play a role in excluding Sertoli cell tumor or yolk sac tumor. Although immunohistochemistry can play an undoubted helpful role in this and selected other areas of testicular tumor evaluation, careful evaluation of the gross and routine microscopic features will solve the vast majority of diagnostic problems. An excellent review of immunohistochemistry in this area by R. E. Emerson, MD, and T. M. Ulbright, MD, is cited herein. Spermatocytic seminoma remains a crucial pitfall in diagnosis, and the pathologist must always be alert to the possible diagnosis when looking at a seminomatous neoplasm, particularly in an older patient, although about one third of these tumors occur in the usual seminoma age range. The embryonal carcinoma has a great diversity of patterns, which are briefly noted. The enigmatic and picturesque tumor, polyembryoma, which virtually never occurs in pure form but may be a confusing component of a variety of mixed germ cell tumors, is discussed and illustrated. The phenomenon of burnt-out germ cell neoplasia is also briefly noted and an excellent recent contribution is referred to. Within the sex cord-stromal family of neoplasms, recent contributions and elaborations of unusual morphologic features of Leydig cell tumors and Sertoli cell tumors are presented. Within the Leydig cell family, cyst formation, adipose metaplasia, calcification or ossification, and spindle cell patterns may be particularly confusing, and in the Sertoli cell family, a great array of patterns caused by differing admixtures of tubular, solid, and stromal components occur. The peculiar lesion, intratubular large cell hyalinizing Sertoli cell tumor, of young boys with Peutz-Jeghers syndrome, is briefly discussed. Some of the problems in the family of hematopoietic neoplasms are reviewed, these processes posing diverse problems in differential diagnosis and their correct recognition having crucial therapeutic implications. Although secondary tumors to the testis have not received the same attention in the literature as the similar phenomenon in the female gonad, remarkable examples of testicular spread of diverse neoplasms, usually carcinoma but rarely melanoma, are seen, and the pathologist should be alert to this possibility, particularly when examining an unusual morphology in an older patient. Finally, a few comments are made on the common paratesticular neoplasm, the adenomatoid tumor, highlighting its varied patterns and recent description of some of the issues that may arise when they undergo total or subtotal infarction. PMID- 18384208 TI - Extranodal lymphoma. AB - Lymphomas arising in extranodal sites are intriguing. The types of lymphomas encountered vary widely from one extranodal site to another. For many types of extranodal lymphomas, there are distinctive clinicopathologic features, sometimes including association with an underlying immunodeficiency syndrome, autoimmune disease, infection, or other immunologic disorder, or a predilection to affect patients of certain ethnic origins. Presented below is a review of lymphomas that are encountered most often in extranodal sites. PMID- 18384209 TI - Pseudosarcomas of soft tissue. AB - One of the most common and important pitfalls in soft tissue pathology are the so called pseudosarcomas. These lesions are nonneoplastic; however, their rapid growth, hypercellularity, cytologic atypia, and mitotic activity makes them prone to be misinterpreted as sarcoma. The most common of these lesions are fibroblastic/myofibroblastic and matrix-forming proliferations, including nodular fasciitis, proliferative fasciitis and myositis, ischemic fasciitis, massive localized edema, myositis ossificans, and bizarre parosteal osteochondromatous proliferation and related entities. Most of these lesions rarely recur following simple excision; therefore, their accurate recognition helps prevent excessive therapy. PMID- 18384210 TI - Reactive versus neoplastic bone marrow: problems and pitfalls. AB - Examination of the bone marrow poses several unique challenges to the pathologist: it is a semisolid organ without helpful gross correlation, it exists in a dynamic state with the peripheral blood and other organs of the lymphohemopoietic system, and the diagnosis of diseases affecting bone marrow often depends heavily on ancillary special studies. The bone marrow examination ideally encompasses review of the bone marrow biopsy histology (with or without additional nondecalcified clot preparation material), bone marrow aspirate smear cytology, and the peripheral blood smear; optimal procurement and processing of these samples is critical in ensuring that a maximal level of diagnostic information can be extracted. The pathologist must be aware of the clinical context of the bone marrow and the results of ancillary tests, whether these are ordered by the pathologist or the clinician. A combination of excellent diagnostic samples, appropriate ancillary tests, and knowledge of the clinical context provides the best background to distinguish between the common reactive and neoplastic processes that involve the bone marrow and to avoid diagnostic pitfalls in making these distinctions. PMID- 18384211 TI - Smooth muscle tumors of the uterus: a practical approach. AB - Smooth muscle tumors (SMTs) are the most frequent mesenchymal tumors of the uterus. The majority of the uterine SMTs are readily classificable as benign or malignant based on their gross and microscopic appearances. However, when unusual features are seen in some leiomyoma variants, the differential diagnosis with a leiomyosarcoma may become challenging. Moreover, diagnostic criteria for the different subtypes of leiomyosarcoma are not uniform. Finally, non-smooth muscle tumors that originate in the uterus may show overlapping histologic and even immunohistochemical features with uterine SMTs, more commonly with the spindle and epithelioid variants, complicating their correct classification. The diagnosis of malignant uterine SMTs has important prognostic and therapeutic implications. This review provides a practical approach to the diagnosis of uterine leiomyosarcoma based on a systematic assessment of histologic parameters as well as a systematic approach to its differential diagnosis based on histologic and immunohistochemical features. PMID- 18384212 TI - The pathology of pulmonary disorders due to Aspergillus spp. AB - Diagnosing the range of pulmonary disorders caused by Aspergillus spp can be challenging. In instances of hypersensitivity responses to Aspergillus spp (ie, allergic bronchopulmonary aspergillosis, bronchocentric granulomatosis, and hypersensitivity pneumonitis), the surgical pathologist must be cognizant of their relationship to infection, as fungal organisms may be rare or absent from the biopsy specimens. Within the spectrum of opportunistic infection, it is critical to distinguish Aspergillus spp from other fungal mimics, as well as to discern whether infection is limited, progressive, or immanently life threatening. However, the surgical pathologist who establishes expertise in this area will be rewarded by the satisfaction of having contributed primarily to an important area of patient care. This article reviews the spectrum of pulmonary disorders due to infection by Aspergillus spp, with emphasis on the clinical implications of diagnosis. PMID- 18384213 TI - Flat epithelial atypia of the breast. AB - Flat epithelial atypia is a presumably neoplastic alteration of terminal duct lobular units that is characterized by the replacement of the native luminal epithelium by ductal cells demonstrating low-grade cytologic atypia. The atypical cells maintain a "flat" pattern of growth without evidence of architectural atypicality. Morphologic, immunohistochemical, and molecular investigations support that flat epithelial atypia represents an early step in the evolution of low-grade ductal carcinomas. It is frequently seen in association with atypical ductal hyperplasia, low-grade ductal carcinoma in situ, invasive tubular carcinoma, and lobular neoplasia. The risk for subsequent breast carcinoma remains to be defined, but flat epithelial atypia likely represents a nonobligate precursor with an extended time course to progression. Certain benign alterations may superficially mimic its appearance; careful attention to cytologic and architectural characteristics can help one distinguish these unrelated entities from flat epithelial atypia. PMID- 18384214 TI - The thyroid gland: recurring problems in histologic and cytologic evaluation. AB - Nodules of the thyroid gland are frequently encountered, occurring in up to 7% of the population, and although most of these nodules are benign, carcinomas of the thyroid gland are the most common malignancy of the endocrine system. Although the different types of thyroid carcinoma are few, a wide variety of recurring problems exists in both their histologic and cytologic evaluation. Here, I will review a selected group of problematic areas, including unusual histologic variants of follicular adenoma, criteria for diagnosing minimally invasive follicular carcinoma, the use of fine-needle aspiration as a screening test for follicular neoplasia, challenging variants of papillary carcinoma, and features of poorly differentiated carcinoma. PMID- 18384215 TI - Gastric polyps: classification and management. AB - Gastric polyps can be broadly defined as luminal lesions projecting above the plane of the mucosal surface. They are relatively frequent in routine pathology practice, where the main goal is to rule out the possibility of malignancy. Various subtypes of gastric polyps are recognized and generally divided into nonneoplastic and neoplastic. We will review herein only a limited subset of gastric polyps representing the most common or, sometimes, challenging. PMID- 18384216 TI - Placental pathology, a survival guide. AB - Placental pathology continues to be an underutilized, undertaught, and inadequately handled surgical subspecialty. The requests for placental pathology are soaring, due partly to demands from obstetricians and to the litigious environment in which they practice, and to improved obstetrical care leading to pregnancies in medically challenging situations. Evaluation of the placenta requires a good understanding of the questions and issues concerning both the fetus/infant and the mother. Information from placental pathology can be critical in early neonatal care and in reproductive planning for the family, and it can provide risk assessment for neurologic outcome of the infant. A comfortable interaction among the obstetric staff, mothers, and pathologists often obviates need for legal intervention in unexpected pregnancy outcomes. Some critical pathologic features that involve maternal and fetal management are illustrated herein. A template for gross examination and a few critical histopathologic diagnostic features with clincopathologic correlation are included. PMID- 18384217 TI - Interface dermatitis. AB - Interface dermatitis can be classified based upon the cell type that dominates the infiltrate (ie, neutrophilic, lymphocytic, or lymphohistiocytic) or by the intensity of the interface inflammation. Regarding lymphocytic interface dermatitis, there are 2 broad categories: cell-poor interface dermatitis, when only a sparse infiltrate of inflammatory cells is present along the dermoepidermal junction, or cell rich, which typically occurs as a heavy bandlike infiltrate that obscures the basal layers of the epidermis. In the case of lymphocytic interface dermatitis, the latter is often termed a lichenoid interface dermatitis. This review focuses upon the mononuclear cell-predominant forms of interface dermatitis. PMID- 18384218 TI - DNA pyrosequencing-based identification of pathogenic Candida species by using the internal transcribed spacer 2 region. AB - CONTEXT: The incidence of infections due to diverse Candida species is increasing, with correspondingly different antifungal susceptibility patterns. Routine yeast identification methods cause significant delays in appropriate patient management. OBJECTIVE: A DNA pyrosequencing strategy was evaluated for identification of pathogenic Candida species associated with human infections. DESIGN: Clinical (n = 51) and commercial (n = 9) Candida isolates were identified in a blinded, parallel study consisting of routine fungal cultures and biochemical analyses in comparison with DNA pyrosequencing. RESULTS: DNA pyrosequencing yielded species-level identification of all 60 Candida isolates, and sequencing interpretations agreed in all cases with results of biochemical and morphologic testing. Different Candida species were identified, such as C. albicans, C. dubliniensis, C. glabrata, C. guilliermondii, C. krusei, C. lusitaniae, C. parapsilosis, and C. tropicalis. Automated and manual approaches to DNA sequence interpretation, each coupled with the Identifire identification software, demonstrated 100% agreement with respect to Candida species identification. Twenty-one isolates yielded intraspecies DNA sequence differences (90%-98% nucleic acid sequence identity) by automated interpretation. Sequence differences resulted from single-nucleotide polymorphisms or single-base additions/deletions, in addition to interpretative challenges in homopolymeric tracts. CONCLUSION: DNA pyrosequencing coupled with automated DNA sequence alignment provides a practical approach for accurate and timely identification of Candida pathogens. Relatively rapid and facile genotypic studies by DNA pyrosequencing matched the effectiveness of extensive biochemical/morphologic studies for yeast identification. PMID- 18384219 TI - Proteomic comparison of nasopharyngeal cancer cell lines C666-1 and NP69 identifies down-regulation of annexin II and beta2-tubulin for nasopharyngeal carcinoma. AB - CONTEXT: Nasopharyngeal carcinoma (NPC), common in southern China and North Africa, has a complex etiology involving interplay between viral, environmental, and hereditary factors and is almost constantly associated with the Epstein-Barr virus. Since the prognosis of locally advanced and metastatic diseases is poor, increased understanding of the pathogenesis of NPC would be important for discovering novel markers for patients' management. OBJECTIVES: To compare the proteomic expression profile between an Epstein-Barr virus-associated NPC cell line (C666-1) and a normal NP cell line (NP69). The proteins with differential expression were analyzed in 40 undifferentiated NPC paraffin-embedded specimens. DESIGN: Differentially expressed proteins discovered between the two cell lines were identified by mass spectrometry. After confirmation by immunocytochemical staining, their expression in patient samples was measured using 40 pairs of undifferentiated NPCs together with their adjacent normal epithelia. RESULTS: Proteomic findings indicated that adenosine triphosphate synthase alpha chain was up-regulated, whereas annexin II, annexin V, beta(2)-tubulin, and profilin 1 were down-regulated. After confirming the results in agar-processed cell lines, annexin II and beta(2)-tubulin expression were found to be lower in tumor cells than in adjacent normal epithelial cells in 100% and 90% of the patients' specimens, respectively. Finally, annexin II down-regulation was positively associated with lymph node metastasis, suggesting that it may be a prognostic factor in NPC. CONCLUSIONS: The results suggest that annexin II and beta(2) tubulin down-regulation is important in NPC formation and may represent potential targets for further investigations. PMID- 18384220 TI - Evaluation of the BiliChek being used on hyperbilirubinemic newborns undergoing home phototherapy. AB - CONTEXT: Newborns are often screened prior to discharge for hyperbilirubinemia. Transcutaneous bilirubin analyzers, such as the BiliChek, are promoted as screening tools, but it is unclear whether they also function well as monitoring devices. Newborns on home phototherapy require frequent determinations of serum bilirubin levels to monitor therapy effects. A transcutaneous bilirubin analyzer would be helpful to limit blood draws and enhance staff efficiency. We evaluated the accuracy of the BiliChek analyzer in this setting. OBJECTIVE: Is the BiliChek sufficiently accurate to monitor the effectiveness of home phototherapy and establish when to terminate therapy? DESIGN: Paired serum bilirubin results and results from the BiliChek were obtained from newborns on home phototherapy during daily home health care visits. RESULTS: The BiliChek demonstrates a negative bias (mean bias, -1.71 mg/dL; 95% confidence interval, -1.89 to -1.52 mg/dL) compared with serum bilirubin values. This bias worsens as the serum bilirubin level rises. If a value of 14 mg/dL or less obtained using the BiliChek had been used as the cutoff for termination of phototherapy, 45% of newborns would have had therapy terminated prematurely. If, knowing the negative bias of the BiliChek, the cutoff for termination of therapy was set at less than or equal to 11 mg/dL, then 29% of newborns would have had therapy terminated prematurely. CONCLUSIONS: The values obtained using the BiliChek, compared to serum bilirubin values, have a negative bias that worsens at the higher bilirubin levels expected in newborns at home on phototherapy. The BiliChek does not provide sufficient accuracy to be utilized to monitor newborns on home phototherapy or to ascertain when to discontinue such therapy. PMID- 18384221 TI - A recapping system for automatic, semiautomatic, and manual use. AB - CONTEXT: Closed sample tubes are needed for stored as well as in-process samples because evaporation significantly changes analyte concentrations. In modern laboratories, it is essential that the procedures for ordering tests on stored serum samples be simple and time-efficient. Therefore, recapping and reopening of tubes should be readily possible in laboratories to save resources and time. OBJECTIVE: To evaluate the recapping system MagCap, in comparison to established snap caps and unsealed tubes. DESIGN: Specific balls, with an iron core, fit tightly by their specific weight into openings of primary and secondary tubes as well as microtiter plates. Recapping of the tubes can be performed individually by a single dispenser or rackwise by a multidispenser. Balls can be removed individually by a pencillike magnet or rackwise. Balls can be reused after cleaning in a glassware washer or used as disposables. RESULTS: Balls seal the sample tubes as effectively as snap caps and decrease evaporation significantly in comparison to unsealed tubes. Evaporation effects of up to 13% were detected in unsealed tubes (0.5 mL) within 24 hours, whereas resealing of the tubes diminished the concentration effect to less than 1%. Open samples (0.5 mL) showed a concentration effect in the refrigerator within 7 days of up to 25%. The evaporation effect of the sealed tubes was less than 3%. Multiple reopenening and recapping procedures are easily possible using magnetic forces. CONCLUSIONS: The MagCap recapping system seals primary and secondary containers securely and cost effectively with the ball cap and thus facilitates a necessary advance in sample quality. PMID- 18384222 TI - The diagnostic value of Ki-67 and repp86 in distinguishing between benign and malignant mesothelial proliferations. AB - CONTEXT: The differentiation of benign mesothelial proliferations from malignant mesotheliomas may be difficult, especially when evaluating small specimens from pleural biopsies. OBJECTIVE: To explore the potential value of 2 proliferative cell markers, Ki-67 and restrictedly expressed proliferation-associated protein 86 kDa (repp86), in distinguishing between malignant mesothelioma (MM) and benign reactive mesothelial hyperplasia (MH). DESIGN: Thirty-six cases of MM from 26 men and 10 women with a mean age of 62.9 years (range, 36-80 years) and 22 cases of benign reactive MH from 14 male and 8 female patients with a mean age of 51.5 years (range, 15- 88 years) were included in this study. The proliferative status of the lesions was assessed by immunohistochemistry using monoclonal antibodies to Ki-S2 (repp86) and Ki-S5 (Ki-67). The labeling indices were quantified. RESULTS: The mean labeling indexes for Ki-67 in MM and benign reactive MH were 24.6% (range, 1%-66%) and 6.23% (range, 0%-25%), respectively. The mean labeling indexes for repp86 in MM and benign reactive MH were 26.3% (range, 0%-50%) and 3.26% (range, 0%- 21%), respectively. The average proliferative cell count was significantly higher in MM compared with benign reactive MH (P < .001). Furthermore, both markers showed a significant correlation in their expression in MM and benign reactive MH (r = 77.5, P < .001). Sensitivities of 88% and 92% and specificities of 92% and 94% were obtained at a cutoff point of 9% for Ki-67 and repp86, respectively. CONCLUSIONS: Used in combination, Ki-67 and repp86 appear to be useful markers in differentiating MM from benign reactive MH. PMID- 18384223 TI - Monophasic synovial sarcoma arising in the vulva: a case report and review of the literature. AB - Synovial sarcomas most commonly arise in the soft tissue of the extremities. Less commonly, these tumors present in the head and neck, abdominal wall, and other sites. However, synovial sarcoma occurring in the vulvar area is extremely rare. Only 2 previous cases of biphasic synovial sarcoma of the vulva have been reported, but no case of vulvar monophasic synovial sarcoma has been described in the English literature. We report the third case of synovial sarcoma and apparently the first case of monophasic synovial sarcoma arising in soft tissues of the vulva. The patient was a 33-year-old woman who presented for evaluation of a painless vulvar mass. The tumor was located in the deep fibroadipose tissue of the right vulva (6.5 x 4.2 x 3.5 cm). The histology of the lesion was that of a monophasic synovial sarcoma with a hemangiopericytic vascular pattern. A subsequent molecular analysis revealed SYT-SSX2 gene fusion, which confirmed the diagnosis of synovial sarcoma. After an initial wide local excision, the patient developed a recurrence in the right groin and received chemotherapy and additional surgery. The patient is currently disease free, on adjuvant chemotherapy, and being followed up closely. PMID- 18384224 TI - Posttransplant adenoviral enteropathy in patients with small bowel transplantation. AB - Transplant patients on immunosuppression represent a risk group for opportunistic infections, including adenoviral infections. The clinical and histopathologic findings of posttransplant adenoviral enteropathy in 3 adult intestinal transplant patients are described. The histopathologic pitfalls for the differential diagnosis between intestinal adenoviral enteropathy and acute rejection are discussed. Adenoviral enteropathy is an opportunistic infection that may follow aggressive treatment for small bowel allograft rejection, but which may require no specific treatment. It is associated with mild mixed inflammation in the lamina propria and slight increase of crypt apoptosis, resembling low-grade acute rejection. The identification of characteristic viral inclusions in the surface or crypt epithelium points to the diagnosis of adenoviral enteropathy, in spite of the increased crypt apoptosis. PMID- 18384225 TI - Epstein-Barr virus-associated lymphoepithelioma-like gastric carcinoma. AB - This article provides an overview of the pathology of Epstein-Barr virus (EBV) associated lymphoepithelioma-like gastric carcinoma, highlighting its unique morphology and clinical features. Lymphoepithelioma-like gastric carcinoma is a rare neoplasm of the stomach with a better prognosis than conventional adenocarcinoma. Most lymphoepithelioma-like gastric carcinomas are associated with EBV infection, while a subset is associated with microsatellite instability. Even though there is a very strong association with EBV, its exact role in carcinogenesis still remains to be elucidated in those cancers that harbor EBV. Distinctive histology and demonstration of EBV using in situ hybridization, polymerase chain reaction, or Southern blotting and immunohistochemistry for the DNA mismatch repair genes or polymerase chain reaction analysis of microsatellite loci to assess microsatellite instability helps to make the diagnosis. PMID- 18384227 TI - Biokinetics of cadmium and zinc in a marine bacterium: influences of metal interaction and pre-exposure. AB - The uptake kinetics of Cd and Zn, as influenced by metal interaction and metal pre-exposure, was examined in the gram-positive marine bacterium Bacillus firmus over a wide range of ambient free-Cd and -Zn concentrations. Bacteria were exposed to experimental media with different concentrations of Cd and Zn over a short, 15-min period. Zinc was found to be an effective competitive inhibitor of Cd uptake when the Zn(2+) concentration ([Zn(2+)]) was increased to 10(-8) M, whereas the Cd concentration (ranging from 10(-9) to 10(-6) M) did not affect Zn uptake. Inhibition of Cd uptake was dependent on [Zn(2+)] instead of the [Zn(2+)] to Cd(2+) concentration ratio. Cadmium uptake at different [Zn(2+)] was significantly inhibited by a sulfur ligand (SH) blocker (N-ethylmaleimide) and a Ca-channel blocker (lanthanum), suggesting that competition between Cd and Zn most likely occurred via binding to the same transport sites. Cadmium efflux also was determined in the presence of different [Zn(2+)]. A biphasic depuration of Cd was found when [Zn(2+)] was greater than 10(-8) M, whereas the calculated Cd efflux rate was independent of [Zn(2+)]. We further exposed B. firmus at different Cd or Zn concentrations for 24 h, then determined the metal uptake and efflux kinetics as well as the metallothionein (MT) induction. Both the Cd and Zn cellular concentrations increased with greater exposed metal concentration, but the MT levels and efflux were little affected by the elevated metal concentration. To some extent, however, the Cd uptake was reduced with an elevated intracellular Zn concentration, suggesting that at high Cd concentrations, intracellular Zn can suppress the Cd uptake in B. firmus. These results help to understand the interactions of metals in the marine environments. PMID- 18384228 TI - Lifetime prevalence of mental disorders in Lebanon: first onset, treatment, and exposure to war. AB - BACKGROUND: There are no published data on national lifetime prevalence and treatment of mental disorders in the Arab region. Furthermore, the effect of war on first onset of disorders has not been addressed previously on a national level, especially in the Arab region. Thus, the current study aims at investigating the lifetime prevalence, treatment, age of onset of mental disorders, and their relationship to war in Lebanon. METHODS AND FINDINGS: The Lebanese Evaluation of the Burden of Ailments and Needs Of the Nation study was carried out on a nationally representative sample of the Lebanese population (n = 2,857 adults). Respondents were interviewed using the fully structured WHO Composite International Diagnostic Interview 3.0. Lifetime prevalence of any Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV) disorder was 25.8%. Anxiety (16.7%) and mood (12.6%) were more common than impulse control (4.4%) and substance (2.2%) disorders. Only a minority of people with any mental disorder ever received professional treatment, with substantial delays (6 to 28 y) between the onset of disorders and onset of treatment. War exposure increased the risk of first onset of anxiety (odds ratio [OR] 5.92, 95% confidence interval [CI] 2.5-14.1), mood (OR 3.32, 95% CI 2.0-5.6), and impulse control disorders (OR 12.72, 95% CI 4.5-35.7). CONCLUSIONS: About one-fourth of the sample (25.8%) met criteria for at least one of the DSM-IV disorders at some point in their lives. There is a substantial unmet need for early identification and treatment. Exposure to war events increases the odds of first onset of mental disorders. PMID- 18384229 TI - Cytological and transcript analyses reveal fat and lazy persister-like bacilli in tuberculous sputum. AB - BACKGROUND: Tuberculous sputum provides a sample of bacilli that must be eliminated by chemotherapy and that may go on to transmit infection. A preliminary observation that Mycobacterium tuberculosis cells contain triacylglycerol lipid bodies in sputum, but not when growing in vitro, led us to investigate the extent of this phenomenon and its physiological basis. METHODS AND FINDINGS: Microscopy-positive sputum samples from the UK and The Gambia were investigated for their content of lipid body-positive mycobacteria by combined Nile red and auramine staining. All samples contained a lipid body-positive population varying from 3% to 86% of the acid-fast bacilli present. The recent finding that triacylglycerol synthase is expressed by mycobacteria when they enter in vitro nonreplicating persistence led us to investigate whether this state was also associated with lipid body formation. We found that, when placed in laboratory conditions inducing nonreplicating persistence, two M. tuberculosis strains had lipid body levels comparable to those found in sputum. We investigated these physiological findings further by comparing the M. tuberculosis transcriptome of growing and nonreplicating persistence cultures with that obtained directly from sputum samples. Although sputum has traditionally been thought to contain actively growing tubercle bacilli, our transcript analyses refute the hypothesis that these cells predominate. Rather, they reinforce the results of the lipid body analyses by revealing transcriptional signatures that can be clearly attributed to slowly replicating or nonreplicating mycobacteria. Finally, the lipid body count was highly correlated (R(2) = 0.64, p < 0.03) with time to positivity in diagnostic liquid cultures, thereby establishing a direct link between this cytological feature and the size of a potential nonreplicating population. CONCLUSION: As nonreplicating tubercle bacilli are tolerant to the cidal action of antibiotics and resistant to multiple stresses, identification of this persister-like population of tubercle bacilli in sputum presents exciting and tractable new opportunities to investigate both responses to chemotherapy and the transmission of tuberculosis. PMID- 18384230 TI - A randomised, blinded, placebo-controlled trial in dementia patients continuing or stopping neuroleptics (the DART-AD trial). AB - BACKGROUND: There have been increasing concerns regarding the safety and efficacy of neuroleptics in people with dementia, but there are very few long-term trials to inform clinical practice. The aim of this study was to determine the impact of long-term treatment with neuroleptic agents upon global cognitive decline and neuropsychiatric symptoms in patients with Alzheimer disease. METHODS AND FINDINGS: DESIGN: Randomised, blinded, placebo-controlled parallel two-group treatment discontinuation trial. SETTING: Oxfordshire, Newcastle and Gateshead, London and Edinburgh, United Kingdom. PARTICIPANTS: Patients currently prescribed the neuroleptics thioridazine, chlorpromazine, haloperidol trifluoperazine or risperidone for behavioural or psychiatric disturbance in dementia for at least 3 mo. INTERVENTIONS: Continue neuroleptic treatment for 12 mo or switch to an identical placebo. OUTCOME MEASURES: Primary outcome was total Severe Impairment Battery (SIB) score. Neuropsychiatric symptoms were evaluated with the Neuropsychiatric Inventory (NPI). RESULTS: 165 patients were randomised (83 to continue treatment and 82 to placebo, i.e., discontinue treatment), of whom 128 (78%) commenced treatment (64 continue/64 placebo). Of those, 26 were lost to follow-up (13 per arm), resulting in 51 patients per arm analysed for the primary outcome. There was no significant difference between the continue treatment and placebo groups in the estimated mean change in SIB scores between baseline and 6 mo; estimated mean difference in deterioration (favouring placebo) -0.4 (95% confidence interval [CI] -6.4 to 5.5), adjusted for baseline value (p = 0.9). For neuropsychiatric symptoms, there was no significant difference between the continue treatment and placebo groups (n = 56 and 53, respectively) in the estimated mean change in NPI scores between baseline and 6 mo; estimated mean difference in deterioration (favouring continue treatment) -2.4 (95% CI -8.2 to 3.5), adjusted for baseline value (p = 0.4). Both results became more pronounced at 12 mo. There was some evidence to suggest that those patients with initial NPI >/= 15 benefited on neuropsychiatric symptoms from continuing treatment. CONCLUSIONS: For most patients with AD, withdrawal of neuroleptics had no overall detrimental effect on functional and cognitive status. Neuroleptics may have some value in the maintenance treatment of more severe neuropsychiatric symptoms, but this benefit must be weighed against the side effects of therapy. TRIAL REGISTRATION: Cochrane Central Registry of Controlled Trials/National Research Register (#ISRCTN33368770). PMID- 18384231 TI - Chile's neoliberal health reform: an assessment and a critique. PMID- 18384232 TI - Improving defences at the portal of HIV entry: mucosal and innate immunity. PMID- 18384233 TI - Exposure to war as a risk factor for mental disorders. PMID- 18384234 TI - Climate change, humans, and the extinction of the woolly mammoth. AB - Woolly mammoths inhabited Eurasia and North America from late Middle Pleistocene (300 ky BP [300,000 years before present]), surviving through different climatic cycles until they vanished in the Holocene (3.6 ky BP). The debate about why the Late Quaternary extinctions occurred has centred upon environmental and human induced effects, or a combination of both. However, testing these two hypotheses climatic and anthropogenic-has been hampered by the difficulty of generating quantitative estimates of the relationship between the contraction of the mammoth's geographical range and each of the two hypotheses. We combined climate envelope models and a population model with explicit treatment of woolly mammoth human interactions to measure the extent to which a combination of climate changes and increased human pressures might have led to the extinction of the species in Eurasia. Climate conditions for woolly mammoths were measured across different time periods: 126 ky BP, 42 ky BP, 30 ky BP, 21 ky BP, and 6 ky BP. We show that suitable climate conditions for the mammoth reduced drastically between the Late Pleistocene and the Holocene, and 90% of its geographical range disappeared between 42 ky BP and 6 ky BP, with the remaining suitable areas in the mid-Holocene being mainly restricted to Arctic Siberia, which is where the latest records of woolly mammoths in continental Asia have been found. Results of the population models also show that the collapse of the climatic niche of the mammoth caused a significant drop in their population size, making woolly mammoths more vulnerable to the increasing hunting pressure from human populations. The coincidence of the disappearance of climatically suitable areas for woolly mammoths and the increase in anthropogenic impacts in the Holocene, the coup de grace, likely set the place and time for the extinction of the woolly mammoth. PMID- 18384235 TI - Chromosomal gene movements reflect the recent origin and biology of therian sex chromosomes. AB - Mammalian sex chromosomes stem from ancestral autosomes and have substantially differentiated. It was shown that X-linked genes have generated duplicate intronless gene copies (retrogenes) on autosomes due to this differentiation. However, the precise driving forces for this out-of-X gene "movement" and its evolutionary onset are not known. Based on expression analyses of male germ-cell populations, we here substantiate and extend the hypothesis that autosomal retrogenes functionally compensate for the silencing of their X-linked housekeeping parental genes during, but also after, male meiotic sex chromosome inactivation (MSCI). Thus, sexually antagonistic forces have not played a major role for the selective fixation of X-derived gene copies in mammals. Our dating analyses reveal that although retrogenes were produced ever since the common mammalian ancestor, selectively driven retrogene export from the X only started later, on the placental mammal (eutherian) and marsupial (metatherian) lineages, respectively. Together, these observations suggest that chromosome-wide MSCI emerged close to the eutherian-marsupial split approximately 180 million years ago. Given that MSCI probably reflects the spread of the recombination barrier between the X and Y, crucial for their differentiation, our data imply that these chromosomes became more widely differentiated only late in the therian ancestor, well after the divergence of the monotreme lineage. Thus, our study also provides strong independent support for the recent notion that our sex chromosomes emerged, not in the common ancestor of all mammals, but rather in the therian ancestor, and therefore are much younger than previously thought. PMID- 18384236 TI - Estimation of the soil-water partition coefficient normalized to organic carbon for ionizable organic chemicals. AB - The sorption of organic electrolytes to soil was investigated. A dataset consisting of 164 electrolytes, composed of 93 acids, 65 bases, and six amphoters, was collected from literature and databases. The partition coefficient log Kow of the neutral molecule and the dissociation constant pKa were calculated by the software ACD/Labs. The Henderson-Hasselbalch equation was applied to calculate dissociation. Regressions were developed to predict separately for the neutral and the ionic molecule species the distribution coefficient (Ka) normalized to organic carbon (Koc) from log Kow and pKa. The log Koc of strong acids (pKa < 4) was not correlated to these parameters. The regressions derived for weak acids and bases (undissociated at environmental pH) were similar. The highest sorption was found for strong bases (pKa > 7.5), probably due to electrical interactions. Nonetheless, their log Koc was highly correlated to log Kow. For bases, a nonlinear regression was developed, too. The new regression equations are applicable in the whole pKa range of acids, bases, and amphoters and are useful in particular for relatively strong bases and amphoters, for which no predictive methods specifically have been developed so far. PMID- 18384237 TI - Distribution and variance/covariance structure of pesticide environmental fate data. AB - Hydrophobicity, persistence, and volatility data for individual pesticides are widely used in risk assessment and transport modeling, so it is important to understand their distribution, variation, and covariation. Correlations (normalized covariance) among properties across a range of multiple pesticides are also important for understanding fundamental relationships among the properties. For the present study, multiple determinations of 11 physicochemical properties of 262 individual pesticides were compiled, primarily from registrant submissions. A Z-score normality analysis indicates that, barring specific data to the contrary, log normality is a reasonable assumption for three properties commonly treated as random variables in modeling: Organic carbon-normalized soil sorption coefficient, aerobic soil metabolism half-life, and field dissipation half-life. Various percentiles for coefficients of variation of the variables are provided, allowing probabilistic modelers to choose realistic population parameters for sampling distributions. A second data set consisting of median values of individual properties for each pesticide was used to investigate the covariance structure of eight of the most important fate properties across 172 pesticides using correlation analysis and exploratory common factor analysis. That analysis demonstrated the use of common factor analysis for reducing the dimensionality of multicollinear environmental fate data, yielding three new orthogonal variables containing most of the information in the original data, and provided insight into the fundamental data structure. PMID- 18384238 TI - Uptake, elimination, and biotransformation of aqueous and dietary DDT in marine fish. AB - To understand the biokinetics and potential risks of p,p' dichlorodiphenyltrichloroethane (DDT) and its metabolites, p,p' dichlorodiphenyldichloroethylene (DDE) and p,p'-dichlorodiphenyldichloroethane (DDD), in fish, we exposed the black sea breams (Acanthopagrus schlegeli) to aqueous and dietary DDTs and then evaluated the bioaccumulation, distribution, biotransformation, and elimination of DDTs under controlled laboratory conditions. The fish rapidly accumulated DDTs from both routes of exposure, particularly the gills and viscera. Elimination of DDTs following aqueous or dietary uptake was slow, and biotranslocation of DDTs was significant during the exposure period but negligible during the depuration period. The biotransformation process was more significant following dietary exposure. During depuration, DDE was the major biotransformed product in the sea breams' carcasses while DDD was the major product in the gills and viscera. However, DDD had a significantly higher elimination rate than DDE and, subsequently, the fish retained more DDE in the body. Intraspecies variability in the elimination and biotransformation processes in fish was observed. We demonstrated that the route of exposure significantly affected the fate and biokinetics of DDTs in fish. The application of a dynamic model provided a tool for quantifying the elimination and biotransformation of DDT in fish. The present study provided insights into the bioaccumulation and biotransformation pathways of DDT in fish that could have important ecotoxicological implications. PMID- 18384239 TI - Global expression profiling of Chlamydomonas reinhardtii exposed to trace levels of free cadmium. AB - In the natural environment, cadmium is often found as a trace contaminant. Due to the complexity of Cd speciation and the heterogeneity of natural systems and processes, it is often difficult to determine clear relationships between analytical measurements of Cd and its induced biological response. Measurements of gene induction can be used to identify molecular mechanisms underlying toxicity and to quantify sublethal responses to trace contaminants. In the present paper, genes that could be involved in the tolerance of Cd to green algae were examined using two global transcriptome profiling strategies. Microarray and differential display techniques were used for a global transcriptome analysis of Chlamydomonas reinhardtii exposed to micromolar and lower Cd(2+) concentrations for a short period (2 h). Real-time quantitative polymerase chain reaction analysis confirmed that a small set of 10 genes was differentially expressed in response to trace Cd(2+) exposures ranging from 7.8 nM to 9.0 microM. Since induction was only observed for a few genes, none of which are known to function in a general stress response, it was likely the result of relevant responses to Cd exposure. The identified genes are discussed with respect to their possible involvement in Cd tolerance and to their future use as biomarkers for monitoring Cd bioavailability in natural soils and waters. PMID- 18384240 TI - Metal distribution and metallothionein induction after cadmium exposure in the terrestrial snail Helix aspersa (Gastropoda, Pulmonata). AB - The aim of the present work was to study the effect of Cd2+ exposure on metallothionein (MT) induction and on the distribution of metals (Cd, Cu, and Zn) in the terrestrial pulmonate Helix aspersa. In particular, the soluble and nonsoluble pools of the accumulated metals and their tissue distribution in uncontaminated and contaminated edible snails were investigated after a two-week exposure to Cd2+. In the soluble cytosolic pool of the midgut gland of H. aspersa, three metal-specific putative MT isoforms were separated following a fractionation protocol with diethylaminoethyl cellulose, size-exclusion chromatography, ultrafiltration, and reversed-phase high-performance liquid chromatography (RP-HPLC). Interestingly, one of the above isoforms seems to bind both Cd and Cu, which may in addition mobilize, after induction by Cd2+, some of the intracellular Cu and, thus, perhaps increase the Cu pool in the cytosolic fraction. The cDNA and its translated amino acid sequence of a Cd2+-binding MT isoform from the snail midgut gland was characterized and attributed to one of the putative MT isoforms obtained by RP-HPLC. The amino acid sequence of this Cd MT isoform of H. aspersa differed from similar sequences described in other terrestrial pulmonates, such as Helix pomatia or Arianta arbustorum, by only a few amino acids (n = 4 and 8, respectively). That the identified Cd-MT from H. aspersa is inducible by Cd2+ also was shown, chromatographic evidence aside, by a specific polymerase chain reaction protocol on a cDNA basis, which included a noninducible housekeeping gene as a control. PMID- 18384244 TI - Missed diagnosis of psychotic depression at 4 academic medical centers. AB - BACKGROUND: Major depressive disorder with psychotic features (psychotic depression), though occurring relatively frequently in the general population, is a commonly missed psychiatric diagnosis. OBJECTIVE: To ascertain accuracy of diagnosis of psychotic depression among inpatients at 4 academic medical centers and explore whether presenting symptoms, treatment setting, and physician's level of training affect the accuracy of diagnosis. METHOD: The medical records of 65 patients who met DSM-IV criteria for psychotic depression following systematic assessment were analyzed to ascertain the concordance between chart diagnoses and research diagnoses arrived at using the Structured Clinical Interview for DSM-IV. The patients were participants in the National Institute of Mental Health Study of Pharmacotherapy of Psychotic Depression, conducted from December 28, 2002, through June 18, 2004, at 4 academic medical centers. For each patient's hospital visit, separate standardized data forms were completed on the basis of each physician's assessment of the patient prior to screening for the study. Hospital records from the emergency room and from admission to psychiatric units were reviewed. Among these 65 patients, 130 chart diagnoses had been made. RESULTS: Psychotic depression had not been diagnosed prior to research assessments for 27% of the 130 diagnoses in our sample. The 3 most common diagnoses assigned to patients meeting research criteria for psychotic depression were major depressive disorder without psychotic features, depression not otherwise specified, and mood disorder not otherwise specified. Failure to identify psychotic depression was more likely when symptoms of depressed mood, hallucinations, or delusions were not noted in the medical record (all p < .005). The accuracy of diagnoses was greater on inpatient units than in emergency rooms (chi(2) = 7.64, p < .01). CONCLUSION: The diagnosis of psychotic depression is frequently missed in emergency room and inpatient settings. The findings of this study are sobering given the serious morbidity and mortality of psychotic depression and the implications for treatment if an inaccurate diagnosis is made. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00056472. PMID- 18384245 TI - Onset of depressive episodes is faster in patients with bipolar versus unipolar depressive disorder: evidence from a retrospective comparative study. AB - OBJECTIVE: Depressive episodes can have a very fast onset (< 1 hour) or start very slowly (> 1 month). This interesting aspect, pointing to different neurophysiological pathomechanisms, has not been systematically evaluated so far. The aim of this study was to describe speed of onset of depressive episodes in a consecutive sample of patients with at least 1 depressive episode and to investigate potential differences between patients with major depression versus bipolar affective disorders concerning this variable. METHOD: We examined 158 consecutive adult patients with major depression (N = 108) and bipolar disorder (N = 50) diagnosed according to criteria of the International Statistical Classification of Diseases, 10th revision, by applying the structured Onset-of Depression Inventory. Patients with acute critical life events preceding the onset were excluded from final analyses. Data were collected between December 2001 and January 2007. RESULTS: There was a significant positive association between speed of onset of the present depressive episode and that of the preceding depressive episode (rho = 0.66, p < .001). Patients with bipolar disorder developed full-blown depressive episodes significantly faster than patients with major depression (p < .001): Whereas depressive episodes began within 1 week in 58% of patients with bipolar disorder, this was the case in only 7.4% of patients with major depression. CONCLUSION: Intraindividually, the speed of onset of depression is similar across different episodes. In the absence of acute critical life events, fast onset of depressive episodes (within 1 week) is common in bipolar disorder but rare in major depression. This aspect might be useful to identify depressive episodes occurring within a bipolar affective illness and might characterize a subgroup of patients with a distinct neurobiology. PMID- 18384246 TI - A double-blind, placebo-controlled study of the opiate antagonist naltrexone in the treatment of pathological gambling urges. AB - OBJECTIVE: Pathological gambling (PG) is a disabling disorder experienced by approximately 1% of adults and for which few empirically validated treatments exist. This study examined the efficacy and tolerability of the opioid antagonist naltrexone in adults with PG who have urges to gamble. METHOD: An 18-week, double blind, placebo-controlled trial was conducted to evaluate the safety and efficacy of 3 doses of oral naltrexone for PG. Seventy-seven individuals with DSM-IV-TR PG were randomly assigned to naltrexone (50 mg/day, 100 mg/day, or 150 mg/day) or placebo. Subjects were assessed with the Pathological Gambling Adaptation of the Yale-Brown Obsessive Compulsive Scale (PG-YBOCS), the urge and behavior subscales of the PG-YBOCS, the Gambling Symptom Assessment Scale (G-SAS), the Clinical Global Impressions-Severity of Illness scale (CGI-S), and measures of depression, anxiety, and psychosocial functioning. Data were collected from September 2002 to June 2005. RESULTS: Outcomes did not significantly differ between the various doses of naltrexone. Subjects assigned to naltrexone had significantly greater reductions in PG-YBOCS total scores (p = .0094), gambling urges (p = .0053), and gambling behavior (p = .0134) compared to subjects assigned to placebo. Subjects assigned to naltrexone also had greater improvement in overall gambling severity (reflected in the CGI-S scores) (p = .0080) and in psychosocial functioning (p = .0177) than subjects assigned to placebo. A complete analysis (N = 49) demonstrated significantly greater improvement on all variables for subjects assigned to naltrexone. A sex analysis demonstrated that men and women did not differ significantly in their response to naltrexone. CONCLUSION: Subjects assigned to naltrexone demonstrated statistically significant reductions in gambling urges and behavior in PG. Low-dose naltrexone (50 mg/day) appeared as efficacious as higher doses (100 mg/day and 150 mg/day), and all doses were well tolerated. PMID- 18384247 TI - A Parent General Behavior Inventory subscale to measure sleep disturbance in pediatric bipolar disorder. AB - OBJECTIVE: Sleep disturbance is a reliable marker for differentiating children with bipolar spectrum disorders from those with attention-deficit/hyperactivity disorder. Sleep-related items were extracted from the Parent General Behavior Inventory (P-GBI) to determine whether these items, as a scale unto themselves, demonstrate adequate psychometrics to be useful as a possible endophenotypic marker for bipolar spectrum disorders. METHOD: From July 2003 to July 2007, 625 youths and their parents completed semistructured Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime version interviews. Parents also completed the 73-item P-GBI. RESULTS: Participants with bipolar spectrum disorders (DSM-IV criteria) scored significantly higher than all other participants on all 7 of the sleep variables (p < .005). On receiver operating characteristic (ROC) analysis, the sleep subscale did substantially as well at discriminating participants with bipolar spectrum disorders as did either of the 2 built-in GBI scales, depression and hypomanic/biphasic (area under the ROC curve = 0.74 vs. 0.75 and 0.77, respectively). CONCLUSION: The P-GBI sleep subscale, developed in this study, is a reliable measure of a wide range of mood related sleep problems in youths diagnosed with bipolar spectrum disorders. Sleep disturbance appears to be a promising endophenotype for further clinical investigation, and the P-GBI sleep scale may provide an inexpensive way of quantifying this trait for research. Further research needs to evaluate how parent report compares to objective measures of sleep efficiency, such as actigraphy or polysomnography. PMID- 18384248 TI - Course of major depressive disorder and suicide outcome: a psychological autopsy study. AB - BACKGROUND: There is considerable debate as to whether suicide is more likely to occur early in the course of major depressive disorder or by cumulative risk, with an increasing risk with each subsequent major depressive episode (MDE). By considering the number of MDEs among representative suicides, we aimed to further investigate the relationship between suicide outcome and the course of major depressive disorder. METHOD: A psychological autopsy method with best informants was used to investigate 154 consecutive suicides who died in the context of a DSM IV MDE. Proxy-based interviews were conducted by using the Structured Clinical Interview for DSM-III-R; the Structured Clinical Interview for DSM-IV Axis II; and a series of behavioral and personality-trait assessments. Second, 143 living depressed outpatients of comparable age to the suicide group were assessed for their history of MDEs. The study was conducted between 2000 and 2005. RESULTS: The distribution of MDEs among depressed suicide completers was as follows: first MDE, 74.7%; second MDE, 18.8%; more than 2 MDEs, 6.5%. This distribution is compared to 32.9% of depressed living outpatients with a single MDE. Increased levels of hostility were associated with single MDE suicide completers. The anxious trait of harm avoidance increased among multiple MDE suicide completers. Alcohol abuse increased among first MDE suicide completers. CONCLUSIONS: Suicide in major depressive disorder is most likely to occur during the first MDE, and this appears to be related to increased levels of the impulsive-aggressive diathesis. PMID- 18384249 TI - Rural parents' perspectives about information on child immunization. AB - INTRODUCTION: Historically, health professionals have used information developed for parents to promote child immunization. Few studies have specifically examined the effectiveness of this information in meeting parents' needs. While the literature emphasizes the importance of clear, thorough, and unbiased information about child immunization, limited attention has been given to what this means from a parent's perspective. The aim of this study was to gain insight in parents' information needs regarding child immunization in order to improve and/or optimize information shared by rural health professionals. We explored: (1) whether any immunization information contributed to parents' decisions; and, if so, how (2) what types of information and content parents required; (3) the sources of information parents considered helpful and trustworthy; and (4) parents' suggestions on how information could be conveyed to them more effectively. METHODS: This was a descriptive qualitative study, using semi structured interviews with legal-aged mothers responsible for decisions about immunizing their infant in the past year. The mothers were from the local rural communities south of Calgary, Alberta, Canada, within the boundaries of the Calgary Health Region. Public health nurses working in this area assisted with recruitment. Thirty-nine mothers expressed interest in the study. The investigator contacted respondents to answer questions they may have had as well as to gather more socio-demographic information. This assisted in drawing a sample that reflected a variety of ages, education levels, and decisions made about immunization. Interviews were conducted by the principal investigator. Data collection and analysis took place simultaneously. Data collection continued until saturation was reached. All three investigators were involved in data analysis and data interpretation to ensure quality of the results. RESULTS: Eleven interviews were conducted. Participants were all mothers, most of whom lived in a stable relationship. Five mothers made the decision to fully immunize their child. The other mothers were varied in their decisions which included waiting to immunize the child until s/he was older, choosing vaccines selectively, being undecided about immunizing, and not immunizing. There were three mothers who had made a different decision about immunization with previous children. Three mothers were first-time parents. Five major themes were identified: (1) factors influencing mothers' decisions; (2) mothers' worries in making their decision; (3) mothers' perceptions about 'good' information; (4) mothers' information needs; and (5) mothers' recommendations to health professionals who convey immunization information to parents. CONCLUSION: The study had some limitations. Only mothers responded to the request for participation and the geographical area of the study was limited to the rural area where those particular public health nurses worked. Participants provided insightful perspectives on the subject of information on child immunization and how that information is conveyed to them. Feedback from the nurses also indicated the results were useful and thought-provoking. Future research in this area, using larger and more diverse populations, would benefit health professionals developing and conveying immunization information to parents. Key words: Canada, decision-making, immunization, infancy and childhood, information needs. PMID- 18384250 TI - Pharmacogenetics: global clinical markers. PMID- 18384252 TI - Toxicogenomics in drug safety assessment. PMID- 18384253 TI - Analysis of the adult human plasma metabolome. AB - OBJECTIVE: It is well established that disease states are associated with biochemical changes (e.g., diabetes/glucose, cardiovascular disease/cholesterol), as are responses to chemical agents (e.g., medications, toxins, xenobiotics). Recently, nontargeted methods have been used to identify the small molecules (metabolites) in a biological sample to uncover many of the biochemical changes associated with a disease state or chemical response. Given that these experimental results may be influenced by the composition of the cohort, in the present study we assessed the effects of age, sex and race on the relative concentrations of small molecules (metabolites) in the blood of healthy adults. METHODS: Using gas- and liquid-chromatography in combination with mass spectrometry, a nontargeted metabolomic analysis was performed on plasma collected from an age- and sex-balanced cohort of 269 individuals. RESULTS: Of the more than 300 unique compounds that were detected, significant changes in the relative concentration of more than 100 metabolites were associated with age. Many fewer differences were associated with sex and fewer still with race. Changes in protein, energy and lipid metabolism, as well as oxidative stress, were observed with increasing age. Tricarboxylic acid intermediates, creatine, essential and nonessential amino acids, urea, ornithine, polyamines and oxidative stress markers (e.g., oxoproline, hippurate) increased with age. Compounds related to lipid metabolism, including fatty acids, carnitine, beta hydroxybutyrate and cholesterol, were lower in the blood of younger individuals. By contrast, relative concentrations of dehydroepiandrosterone-sulfate (a proposed antiaging androgen) were lowest in the oldest age group. Certain xenobiotics (e.g., caffeine) were higher in older subjects, possibly reflecting decreases in hepatic cytochrome P450 activity. CONCLUSIONS: Our nontargeted analytical approach detected a large number of metabolites, including those that were found to be statistically altered with age, sex or race. Age-associated changes were more pronounced than those related to differences in sex or race in the population group we studied. Age, sex and race can be confounding factors when comparing different groups in clinical studies. Future studies to determine the influence of diet, lifestyle and medication are also warranted. PMID- 18384254 TI - Filaggrin loss-of-function mutations and association with allergic diseases. AB - Human skin constitutes a highly organized barrier against environmental agents. Its unrestricted function depends on a complex interplay between multiple proteins and lipids expressed in the terminally differentiating epithelium. Recently, attention has been drawn to the protein filaggrin, an integral part of the epidermis that plays a key role in engineering and maintaining the barrier function. Common loss-of-function mutations within the filaggrin gene have been demonstrated to cause ichthyosis vulgaris, one of the most common heritable disorders of cornification, and to represent major risk factors for atopic eczema and secondary allergic diseases. The observations on filaggrin provide striking new insights into the etiology of atopic diseases and might pave the way for the development of new therapeutic approaches. PMID- 18384255 TI - Polymorphism in human organic cation transporters and metformin action. AB - Considerable interindividual variabilities in clinical efficacy and adverse events are sometimes recognized in the treatment of Type 2 diabetes mellitus with oral antihyperglycemic drugs. Metformin is the most commonly used biguanide in clinical practice, and also improves insulin resistance and reduces cardiovascular risk. However, certain patients taking metformin do not respond sufficiently. The molecular reasons for the variability in response to metformin are not clear. However, it has been recently suggested that genetic factors may be responsible for the variability. Metformin is not metabolized but is transported by at least two organic cation transporters (OCT), OCT1 and OCT2. Recently, genetic polymorphisms in OCT 1 and OCT2 have been found to be associated with changes in pharmacokinetic/pharmacodynamic responses to substrate drugs. This review focuses on the impact of the genetic polymorphism of organic cation transporters on transport activity, and the implications for the clinical efficacy of metformin. PMID- 18384256 TI - Genetic variations and their influence on risk and treatment of venous thrombosis. AB - Venous thrombosis (VT) is a highly prevalent disease. Risk factors can be genetic or acquired. The well-established genetic polymorphisms predisposing to thrombophilic disorders can be divided into rare 'loss-of-function mutations' in anticoagulant proteins and common 'gain-of-function mutations' in procoagulant proteins, which are weaker risk factors. In addition to functional polymorphisms, defects in common pathways affecting biosynthesis or clearance of plasma coagulation factors and their relations to VT risk have been detected. Recently, investigations regarding genetic variations and response to drug treatment, relevant for the pathogenesis as well as therapy of venous thromboembolism have been performed. The methodical advances in genetic research have led to the identification of a number of new variants with still unclear association to VT. This review aims to discuss the established genetic risk factors as well as some candidate predictors of VT. Further, the recent developments in pharmacogenomics are reviewed. PMID- 18384257 TI - An update on methotrexate pharmacogenetics in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a systemic inflammatory disorder that mainly affects the joints. When left untreated, the disease can result in irreversible joint damage with high morbidity and mortality. Disease-modifying antirheumatic drugs are the cornerstones of treatment in RA. Disease-modifying antirheumatic drugs not only ameliorate the clinical signs and symptoms of disease, but also prevent the radiographic progression of joint damage. Methotrexate is one such disease modifying antirheumatic drug that has been used in the treatment of RA for over two decades with excellent long-term efficacy and safety. However, there is significant variability in patients' response to methotrexate, both in terms of efficacy and toxicity. At the present time, there are no reliable means of predicting, a priori, an individual patient's response to methotrexate. In this review, recent published literature on the pharmacogenetics of methotrexate in RA is highlighted. Pharmacogenetics may be a powerful tool for optimizing methotrexate therapy in patients with RA. PMID- 18384259 TI - DxS Ltd. AB - DxS is a personalized medicine company that meets the needs of the pharmaceutical industry for biomarkers and companion diagnostics to support the development and sales of cancer and other therapies. The company provides both biomarker products, which are used predominately during clinical trials, and companion diagnostics, which aid doctors in selecting therapies for patients. Working in partnership with drug companies, DxS offer validated biomarker assays to support drug development and then regulatory approval, by identifying likely responders to drug therapies. DxS have launched the world's first cancer mutation companion diagnostic to support Amgen's Vectibix colorectal cancer therapy. DxS kits detect mutations in oncogenes associated with cancer drug response. TheraScreen is the range of CE-marked diagnostic products for detecting mutations in the EGFR and K RAS genes. Validated biomarker kits are available for research use for EGFR, RAS, RAF, BCR-ABL and other genes that show a correlation between patient mutation status and drug response. PMID- 18384260 TI - Genelex Corporation. AB - Comprehensive, personalized medication management and pharmacogenetic testing are important existing opportunities to reduce adverse medication events and improve overall healthcare outcomes. A primary barrier to the adoption of personalized pharmacology is the inadequacy of existing patient records, drug interaction tools and the 'interpretation gap'--the lack of physician decision support tools needed to interpret DNA test reports. GeneMedRx, an algorithm-driven, gene-drug interaction software, closes this gap. It helps physicians optimize medication regimens by correlating the genetic makeup of the patient with all the medicines they are taking. Portable personal health records created by GeneMedRx are the core product required for a much-needed comprehensive program of personalized medication management. PMID- 18384258 TI - Race, ethnicity and social class and the complex etiologies of asthma. AB - Asthma is a common but complex respiratory disease caused by the interaction of genetic and environmental factors. Significant racial and ethnic disparities in prevalence, mortality and drug response have been described. These disparities may be explained by racial and ethnic-specific variation in genetic, environmental, social and psychological risk factors. In addition, race, ethnicity and social class are important proxies for unmeasured factors that influence health outcomes. Herein, we review salient differences in the etiologies of asthma by race, ethnicity and social class, and argue for their continued use as variables in asthma research. PMID- 18384261 TI - Helicos BioSciences. AB - Helicos BioSciences Corporation is a life sciences company developing revolutionary new single molecule sequencing technology to provide the path to the US$1000 genome. True Single Molecule Sequencing (tSMS) will drive advancements in pharmacogenomics that can enable a better understanding of an individual's susceptibility to disease, develop more effective disease diagnoses and differentiate response to disease therapies. During 2007, genome-wide disease association studies, the encylopedia of DNA elements (ENCODE) and the published genome sequence of two individuals have revealed human genome variation far more extensive than originally believed. These also demonstrated that common variations explain only a fraction of the genetic basis of disease. Therefore, the capability to understand an individual genome is critical in setting the foundation for the next great revolution in healthcare. Helicos is committed to this vision and will provide cost-effective genome sequencing and comprehensive analysis of the transcribed genome that can unlock the era of personalized healthcare. PMID- 18384263 TI - Correcting the estimated level of differential expression for gene selection bias: application to a microarray study. AB - The level of differential gene expression may be defined as a fold change, a frequency of upregulation, or some other measure of the degree or extent of a difference in expression across groups of interest. On the basis of expression data for hundreds or thousands of genes, inferring which genes are differentially expressed or ranking genes in order of priority introduces a bias in estimates of their differential expression levels. A previous correction of this feature selection bias suffers from a lack of generality in the method of ranking genes, from requiring many biological replicates, and from unnecessarily overcompensating for the bias. For any method of ranking genes on the basis of gene expression measured for as few as three biological replicates, a simple leave-one-out algorithm corrects, with less overcompensation, the bias in estimates of the level of differential gene expression. In a microarray data set, the bias correction reduces estimates of the probability of upregulation or downregulation from 100% to as low as 60%, even for genes with estimated local false discovery rates close to 0. A simulation study quantifies both the advantage of smoothing estimates of bias before correction and the degree of overcompensation. PMID- 18384264 TI - TRAB: testing whether mutation frequencies are above an unknown background. AB - To rigorously determine whether a gene or a set of genes have alterations that are involved in carcinogenesis requires a comparison of the prevalence of identified changes to a control mutation frequency present in tumor DNA. To facilitate this task, we develop a testing approach and the associated R library, called TRAB, that evaluates whether the frequency of somatic mutation in a given gene is higher than that observed in a control group of genes. Specifically, we test the null hypothesis that the frequency belongs to a control population of frequencies, against the alternative hypothesis that the frequency is higher. Mutation frequencies in the control group are themselves allowed to be variable. TRAB computes the a posteriori probability and the Bayes factor for the hypothesis using a hierarchical Bayesian approach. PMID- 18384265 TI - Adapting prediction error estimates for biased complexity selection in high dimensional bootstrap samples. AB - The bootstrap is a tool that allows for efficient evaluation of prediction performance of statistical techniques without having to set aside data for validation. This is especially important for high-dimensional data, e.g., arising from microarrays, because there the number of observations is often limited. For avoiding overoptimism the statistical technique to be evaluated has to be applied to every bootstrap sample in the same manner it would be used on new data. This includes a selection of complexity, e.g., the number of boosting steps for gradient boosting algorithms. Using the latter, we demonstrate in a simulation study that complexity selection in conventional bootstrap samples, drawn with replacement, is severely biased in many scenarios. This translates into a considerable bias of prediction error estimates, often underestimating the amount of information that can be extracted from high-dimensional data. Potential remedies for this complexity selection bias, such as alternatively using a fixed level of complexity or of using sampling without replacement are investigated and it is shown that the latter works well in many settings. We focus on high dimensional binary response data, with bootstrap .632+ estimates of the Brier score for performance evaluation, and censored time-to-event data with .632+ prediction error curve estimates. The latter, with the modified bootstrap procedure, is then applied to an example with microarray data from patients with diffuse large B-cell lymphoma. PMID- 18384266 TI - Adaptive choice of the number of bootstrap samples in large scale multiple testing. AB - It is a common practice to use resampling methods such as the bootstrap for calculating the p-value for each test when performing large scale multiple testing. The precision of the bootstrap p-values and that of the false discovery rate (FDR) relies on the number of bootstraps used for testing each hypothesis. Clearly, the larger the number of bootstraps the better the precision. However, the required number of bootstraps can be computationally burdensome, and it multiplies the number of tests to be performed. Further adding to the computational challenge is that in some applications the calculation of the test statistic itself may require considerable computation time. As technology improves one can expect the dimension of the problem to increase as well. For instance, during the early days of microarray technology, the number of probes on a cDNA chip was less than 10,000. Now the Affymetrix chips come with over 50,000 probes per chip. Motivated by this important need, we developed a simple adaptive bootstrap methodology for large scale multiple testing, which reduces the total number of bootstrap calculations while ensuring the control of the FDR. The proposed algorithm results in a substantial reduction in the number of bootstrap samples. Based on a simulation study we found that, relative to the number of bootstraps required for the Benjamini-Hochberg (BH) procedure, the standard FDR methodology which was the proposed methodology achieved a very substantial reduction in the number of bootstraps. In some cases the new algorithm required as little as 1/6th the number of bootstraps as the conventional BH procedure. Thus, if the conventional BH procedure used 1,000 bootstraps, then the proposed method required only 160 bootstraps. This methodology has been implemented for time-course/dose-response data in our software, ORIOGEN, which is available from the authors upon request. PMID- 18384267 TI - A cross-validation study to select a classification procedure for clinical diagnosis based on proteomic mass spectrometry. AB - We present an approach to construct a classification rule based on the mass spectrometry data provided by the organizers of the "Classification Competition on Clinical Mass Spectrometry Proteomic Diagnosis Data." Before constructing a classification rule, we attempted to pre-process the data and to select features of the spectra that were likely due to true biological signals (i.e., peptides/proteins). As a result, we selected a set of 92 features. To construct the classification rule, we considered eight methods for selecting a subset of the features, combined with seven classification methods. The performance of the resulting 56 combinations was evaluated by using a cross-validation procedure with 1000 re-sampled data sets. The best result, as indicated by the lowest overall misclassification rate, was obtained by using the whole set of 92 features as the input for a support-vector machine (SVM) with a linear kernel. This method was therefore used to construct the classification rule. For the training data set, the total error rate for the classification rule, as estimated by using leave-one-out cross-validation, was equal to 0.16, with the sensitivity and specificity equal to 0.87 and 0.82, respectively. PMID- 18384268 TI - Integrating spirituality into undergraduate nursing curricula. AB - Nursing programs have done a commendable job keeping pace with the rapid advances in disease management. Yet, spirituality has received far less attention in nursing curricula (Keefe, 2005) and nursing students often do not have a strong foundation in this area. The purpose of this project was to integrate spirituality into the undergraduate nursing curricula and measure student outcomes related to spiritual knowledge and attitudes. Nursing faculty participated in a spirituality education program and followed this with sessions focused on integration of spiritual content into individual nursing courses. Student pre and post-tests were administered using a standard instrument to evaluate the effectiveness of the program. Significant differences in spirituality knowledge and attitudes among senior-level nursing students (t = 3.059, p = .004) were revealed. As the healthcare system becomes increasingly complex, providing students with tools to identify and strengthen inner resources is essential to patient care. PMID- 18384269 TI - The influence of end-of-life education on attitudes of nursing students. AB - Palliative care is an important aspect of nursing when comfort and quality of life are the patient goals. The End-of-Life Nursing Education Consortium (ELNEC) developed a comprehensive program of teaching care of the dying to nurses and nursing students. This pretest-posttest study evaluated the influence of the integration of the ELNEC curriculum into a baccalaureate nursing program on students' attitudes toward care of the dying. The Frommelt Attitudes toward Care of the Dying Scale for nurses (FATCOD) was administered to traditional and accelerated baccalaureate students before and after exposure to a nursing curriculum that integrated essential ELNEC elements. Multiple regression analyses indicated that no previous experience with death and an age of 18-22 accounted for the most variance in attitude change. The findings suggest that integrating the ELNEC curriculum throughout a baccalaureate program positively affects the attitudes of nursing students toward the care of patients who are dying. PMID- 18384270 TI - Limited use of the human patient simulator by nurse faculty: an intervention program designed to increase use. AB - Underutilization of human patient simulators (HPS) is not only a curricular issue but also a resource allocation problem. The study explored factors contributing to the limited HPS by faculty in a large ADN program. There is limited empirical evidence published to address this phenomenon. The researchers surveyed the faculty to identify their beliefs and challenges with implementing simulation based upon the constructs (attitudes, subjective norms, perceived behavioral control and intent to use) of the Theory of Planned Behavior (TPB). An educational intervention to address these specific challenges was implemented. The intervention had a positive influence on all TPB construct means (attitudes, p < .01; subjective norms, p < .01; perceived behavioral control, p < .01; intent, p < .05). Attitude (beta = .896) was the strongest predictor in explaining intent to use the HPS. This evidenced-based study offers an approach to increasing the use of the HPS by faculty. PMID- 18384271 TI - Diffusion of innovation: embedding simulation into nursing curricula. AB - Numerous articles have documented the benefits of using simulation as a teaching method for undergraduate nursing students. Simulation can enhance learning and provides a stimulating environment for technologically proficient students. Yet, there remain a large number of nursing programs and faculty members that are resistant toward implementing simulation as a learning tool. This article provides details on the efforts to embed simulation in an undergraduate program that started with a few interested faculty at a large, multi-site nursing program. The Diffusion of Innovation theory was used to guide the expansion of simulation to other faculty groups. The techniques used to embed simulation into the undergraduate curriculum were directed by past research. This process led to a successful integration of simulation which could provide some innovative suggestions for other programs facing similar barriers. PMID- 18384272 TI - Online post LPN to BN students' views of transitioning to a new nursing role. AB - Vocationally prepared nurses, most of whom are women, experience unique challenges when they enter an online university. And yet, educational research examining their perceptions about transitioning into a new professional role is limited. This qualitative descriptive study investigated beginning Post LPN to BN students' views of role differences among nurses and the process of transitioning between those roles. The project was framed from a constructivist theoretical perspective and applied repertory grid methodology from the field of personal construct psychology. Ten students from a Canadian online university were interviewed. Interview transcripts were analyzed for themes. Findings revealed that students appreciated recognition for their previous accomplishments. They valued affirmation of the unique challenges they faced. And, they associated gains from their new university education with some loss of their hands on bedside nursing role. PMID- 18384273 TI - Nursing student-patient relationships: a descriptive study of students' and patients' views. AB - This study was designed to describe and compare nursing students' and patients' perceptions of the relationship between students and patients. The data was collected in Finland from a convenience sample of nursing students (n=290) and patients (n=242) using questionnaires especially designed for this study. The results indicated that students viewed the relationship as more authoritative and facilitative, while patients regarded the relationship as more mechanistic. Furthermore, students' and patients' views on their relationships differed significantly. These results have important implications for nursing education. They particularly highlight the need to further strengthen nursing student patient interactions and relationships. PMID- 18384274 TI - Comparing NET and ERI standardized exam scores between baccalaureate graduates who pass or fail the NCLEX-RN. AB - In the United States, nursing programs are commonly evaluated by their graduates success on the National Council Licensure Examination for Registered Nurses (NCLEX-RN). The purpose of this paper is to describe a change in NCLEX-RN success rates following the addition of standardized exams throughout our program's curriculum, and to compare these exam scores between graduates who pass NCLEX-RN and those who do not. Our results indicate an 8.5% change (p < 0.000) in the NCLEX-RN pass rate from our previous 5-year mean pass rate, and significant differences in standardized test scores for those who pass the NCLEX-RN compared to those who do not (p < 0.03). We conclude that our selected standardized exam scores are able to significantly identify graduates who are more likely to pass NCLEX-RN than not. PMID- 18384275 TI - "Failure to assign failing grades": issues with grading the unsafe student. AB - In a grounded theory study examining the process of precepting an unsafe student, it was found that preceptors assigned passing grades to students who in fact should not have passed. Although preceptors perceived their role as gatekeepers for the profession, by not assigning failing grades to students who should not have passed a course, essentially they were abdicating their responsibility. Indeed, the simple act of assuming responsibility for precepting a student implies professional as well as pedagogical accountability. PMID- 18384276 TI - Factors influencing intent to stay in academia for nursing faculty in the southern United States of America. AB - The current nursing faculty shortage makes understanding intent to stay in academia a step toward slowing the exodus of faculty. The purpose of this study was to discover a parsimonious set of predictor variables from the variables of job satisfaction, mentoring, organizational commitment, and leadership behaviors for intent to stay in nursing education. A random cluster sample consisted of 39 schools of nursing in states within the Southern Regional Education Board (SREB). There were 316 responses from 782 potential participants for a response rate of 40.4%. Stepwise multiple regression results indicated that organizational commitment explained 19.7% of the variance in intent to stay one year and 21.2% of the variance in intent to stay five years. Further, mentored faculty scored significantly higher on organizational commitment. Intent to stay three years was not significantly correlated to any predictor variables suggesting it is a time to intervene to influence decisions to stay. PMID- 18384277 TI - Relationships of body mass index with serum carotenoids, tocopherols and retinol at steady-state and in response to a carotenoid-rich vegetable diet intervention in Filipino schoolchildren. AB - In marginally nourished children, information is scarce regarding the circulating concentrations of carotenoids and tocopherols, and physiological factors influencing their circulating levels. We determined the serum concentrations of carotenoids, tocopherols and retinol at steady state and in response to a 9-week vegetable diet intervention in 9-12-year-old girls (n=54) and boys (n=65) in rural Philippines. We determined cross-sectional relationships of BMI (body mass index) with serum micronutrient levels, and whether BMI is a determinant of serum carotenoid responses to the ingestion of carotenoid-rich vegetables. We measured dietary nutrient intakes and assessed inflammation by measurement of serum C reactive protein levels. The children had low serum concentrations of carotenoids, tocopherols and retinol as compared with published values for similar-aged children in the U.S.A. The low serum retinol levels can be ascribed to inadequate diets and were not the result of confounding due to inflammation. Significant inverse correlations of BMI and serum all-trans-beta-carotene, 13-cis beta-carotene, alpha-carotene, lutein, zeaxanthin and alpha-tocopherol (but not beta-cryptoxanthin, lycopene and retinol) were observed among girls at baseline. The dietary intervention markedly enhanced the serum concentrations of all carotenoids. Changes in serum all-trans-beta-carotene and alpha-carotene (but not changes in lutein, zeaxanthin and beta-cryptoxanthin) in response to the dietary intervention were inversely associated with BMI in girls and boys. Thus, in Filipino school-aged children, BMI is inversely related to the steady-state serum concentrations of certain carotenoids and vitamin E, but not vitamin A, and is a determinant of serum beta- and alpha-carotene responses, but not xanthophyll responses, to the ingestion of carotenoid-rich vegetable meals. PMID- 18384278 TI - Stimulation of mitochondrial proton conductance by hydroxynonenal requires a high membrane potential. AB - Mild uncoupling of oxidative phosphorylation, caused by a leak of protons back into the matrix, limits mitochondrial production of ROS (reactive oxygen species). This proton leak can be induced by the lipid peroxidation products of ROS, such as HNE (4-hydroxynonenal). HNE activates uncoupling proteins (UCP1, UCP2 and UCP3) and ANT (adenine nucleotide translocase), thereby providing a negative feedback loop. The mechanism of activation and the conditions necessary to induce uncoupling by HNE are unclear. We have found that activation of proton leak by HNE in rat and mouse skeletal muscle mitochondria is dependent on incubation with respiratory substrate. In the presence of HNE, mitochondria energized with succinate became progressively more leaky to protons over time compared with mitochondria in the absence of either HNE or succinate. Energized mitochondria must attain a high membrane potential to allow HNE to activate uncoupling: a drop of 10-20 mV from the resting value is sufficient to blunt induction of proton leak by HNE. Uncoupling occurs through UCP3 (11%), ANT (64%) and other pathways (25%). Our findings have shown that exogenous HNE only activates uncoupling at high membrane potential. These results suggest that both endogenous HNE production and high membrane potential are required before mild uncoupling will be triggered to attenuate mitochondrial ROS production. PMID- 18384279 TI - Drugs and ergogenic aids to improve sport performance. PMID- 18384280 TI - The limits of human performance. AB - Human performance, defined by mechanical resistance and distance per time, includes human, task and environmental factors, all interrelated. It requires metabolic energy provided by anaerobic and aerobic metabolic energy sources. These sources have specific limitations in the capacity and rate to provide re phosphorylation energy, which determines individual ratios of aerobic and anaerobic metabolic power and their sustainability. In healthy athletes, limits to provide and utilize metabolic energy are multifactorial, carefully matched and include a safety margin imposed in order to protect the integrity of the human organism under maximal effort. Perception of afferent input associated with effort leads to conscious or unconscious decisions to modulate or terminate performance; however, the underlying mechanisms of cerebral control are not fully understood. The idea to move borders of performance with the help of biochemicals is two millennia old. Biochemical findings resulted in highly effective substances widely used to increase performance in daily life, during preparation for sport events and during competition, but many of them must be considered as doping and therefore illegal. Supplements and food have ergogenic potential; however, numerous concepts are controversially discussed with respect to legality and particularly evidence in terms of usefulness and risks. The effect of evidence-based nutritional strategies on adaptations in terms of gene and protein expression that occur in skeletal muscle during and after exercise training sessions is widely unknown. Biochemical research is essential for better understanding of the basic mechanisms causing fatigue and the regulation of the dynamic adaptation to physical and mental training. PMID- 18384281 TI - Legal pre-event nutritional supplements to assist energy metabolism. AB - Physical training and proper nutrition are paramount for success in sport. A key tissue is skeletal muscle, as the metabolic pathways that produce energy or ATP allow the muscles to complete the many activities critical to success in sport. The energy-producing pathways must rapidly respond to the need for ATP during sport and produce energy at a faster rate or for a longer duration through training and proper nutrition which should translate into improved performance in sport activities. There is also continual interest in the possibility that nutritional supplements could further improve muscle metabolism and the provision of energy during sport. Most legal sports supplements do not improve performance following oral ingestion. However, three legal supplements that have received significant attention over the years include creatine, carnitine and sodium bicarbonate. The ingestion of large amounts of creatine for 4-6 days increases skeletal muscle creatine and phosphocreatine contents. The majority of the experimental evidence suggests that creatine supplementation can improve short term exercise performance, especially in sports that require repeated short-term sprints. It may also augment the accretion of skeletal muscle when taken in combination with a resistance-exercise training programme. Supplementary carnitine has been touted to increase the uptake and oxidation of fat in the mitochondria. However, muscle carnitine levels are not augmented following oral carnitine supplementation and the majority of well-controlled studies have reported no effect of carnitine on enhancing fat oxidation, Vo(2max) or prolonged endurance exercise performance. The ingestion of sodium bicarbonate before intense exercise decreases the blood [H+] to potentially assist the efflux of H+ from the muscle and temper the metabolic acidosis associated with intense exercise. Many studies have reported performance increases in laboratory-based cycling tests and simulated running races in the field following sodium bicarbonate ingestion where the need for ATP from substrate phosphorylation is high. However, other studies have reported no benefit and the incidence of negative side effects is high. PMID- 18384282 TI - Legal nutritional supplements during a sporting event. AB - Nutrition significantly influences sports performance; however, the efficacy of any nutritional supplement or strategy should be carefully considered in relation to the event and the sex, training and nutritional status of the participant. The causes of fatigue, mechanism of action, safety and legality of the supplement, together with the scientific evidence from studies with an appropriate experimental design, should all be taken into account before incorporating into the training and/or competition diet. The efficacy of ingesting nutritional supplements immediately before and/or during endurance exercise (duration 45-180 min) is reviewed in this chapter. The ingestion of CES (carbohydrate-electrolyte solutions) have been shown to improve both exercise capacity and performance, either due to the maintenance of euglycaemia throughout exercise or the sparing of muscle glycogen early on in exercise. The addition of caffeine to CES may improve endurance performance as a consequence of a reduced perception of effort. Research suggests that the addition of protein to CES may only be effective when a suboptimal amount of CHO (carbohydrate) is ingested during exercise (<60 g of CHO.h(-1)); however, recovery of performance may be enhanced due to a reduction in subsequent muscle soreness and the promotion of muscle protein synthesis after exercise. The findings from studies investigating the effects of ingesting MCTs (medium-chain triacylglycerols) and BCAAs (branched-chain amino acids), either on their own or in combination with CES, on endurance performance have been equivocal and therefore would not be recommended. Any nutritional strategy should be practised in training before being used during a competition. PMID- 18384283 TI - The biochemistry of drugs and doping methods used to enhance aerobic sport performance. AB - Optimum performance in aerobic sports performance requires an efficient delivery to, and consumption of, oxygen by the exercising muscle. It is probable that maximal oxygen uptake in the athlete is multifactorial, being shared between cardiac output, blood oxygen content, muscle blood flow, oxygen diffusion from the blood to the cell and mitochondrial content. Of these, raising the blood oxygen content by raising the haematocrit is the simplest acute method to increase oxygen delivery and improve sport performance. Legal means of raising haematocrit include altitude training and hypoxic tents. Illegal means include blood doping and the administration of EPO (erythropoietin). The ability to make EPO by genetic means has resulted in an increase in its availability and use, although it is probable that recent testing methods may have had some impact. Less widely used illegal methods include the use of artificial blood oxygen carriers (the so-called 'blood substitutes'). In principle these molecules could enhance aerobic sports performance; however, they would be readily detectable in urine and blood tests. An alternative to increasing the blood oxygen content is to increase the amount of oxygen that haemoglobin can deliver. It is possible to do this by using compounds that right-shift the haemoglobin dissociation curve (e.g. RSR13). There is a compromise between improving oxygen delivery at the muscle and losing oxygen uptake at the lung and it is unclear whether these reagents would enhance the performance of elite athletes. However, given the proven success of blood doping and EPO, attempts to manipulate these pathways are likely to lead to an ongoing battle between the athlete and the drug testers. PMID- 18384284 TI - Improving muscle mass: response of muscle metabolism to exercise, nutrition and anabolic agents. AB - Muscle mass is critical for athletic performance and, perhaps more importantly for most, health and survival. The metabolic basis for a change in muscle mass is an increase in net muscle protein balance (termed NBAL). NBAL is the difference between MPS (muscle protein synthesis) and MPB (muscle protein breakdown). Thus an increase in MPS and/or a decrease in MPB are necessary for NBAL to increase, leading to accretion of muscle proteins. In particular, accretion of myofibrillar proteins is necessary. NBAL responds to exercise, feeding and other factors. In healthy, weight-stable adults, muscle mass remains constant because periods of positive balance following feeding are countered by periods of negative balance during fasting. A combination of resistance exercise and nutrition is a potent anabolic stimulus through stimulation of MPS from amino acids and attenuation of MPB by carbohydrates. Increased muscle mass results from the accumulation of small amounts of protein in response to each bout of exercise combined with nutrient intake. The magnitude of the response may be influenced by factors other than just the amount of a nutrient ingested. Timing of ingestion, co-ingestion of nutrients and the type of protein may all influence protein accretion. Testosterone is a potent anabolic stimulus primarily through improvement in re utilization of amino acids from MPB. There is a general lack of efficacy in studies assessing the potential for growth hormone, androstenedione and dehydroepiandrostenedione to increase muscle mass. Creatine supplementation is clearly an effective means to increase muscle mass, especially in combination with resistance exercise, however the mechanisms remain unclear. Results from acute metabolic studies provide useful information for estimation of the efficacy of anabolic agents. PMID- 18384285 TI - Signal transduction pathways that regulate muscle growth. AB - Progressive high-resistance exercise with 8-12 repetitions per set to near failure for beginners and 1-12 repetitions for athletes will increase muscle protein synthesis for up to 72 h; approx. 20 g of protein, especially when ingested directly after exercise, will promote high growth by elevating protein synthesis above breakdown. Muscle growth is regulated by signal transduction pathways that sense and compute local and systemic signals and regulate various cellular functions. The main signalling mechanisms are the phosphorylation of serine, threonine and tyrosine residues by kinases and their dephosphorylation by phosphatases. Muscle growth is stimulated by the mTOR (mammalian target of rapamycin) system, which senses (i) IGF-1 (insulin-like growth factor 1)/MGF (mechano-growth factor)/insulin and/or (ii) mechanical signals, (iii) amino acids and (iv) the energetic state of the muscle, and regulates protein synthesis accordingly. The action of the mTOR system is opposed by myostatin-Smad signalling which inhibits muscle growth via gene transcription. PMID- 18384286 TI - Caffeine and other sympathomimetic stimulants: modes of action and effects on sports performance. AB - Stimulants, illegal and legal, continue to be used in competitive sport. The evidence for the ergogenic properties of the most potent stimulants, amphetamines, cocaine and ephedrine, is mostly insubstantial. Low doses of amphetamines may aid performance where effects of fatigue adversely affect higher psychomotor activity. Pseudoephedrine, at high doses, has been suggested to improve high intensity and endurance exercise but phenylpropanolamine has not been proven to be ergogenic. Only caffeine has substantial experimental backing for being ergogenic in exercise. The mode of action of these stimulants centres on their ability to cause persistence of catecholamine neurotransmitters, with the exception of caffeine which is an adenosine receptor antagonist. By these actions, the stimulants are able to influence the activity of neuronal control pathways in the central (and peripheral) nervous system. Rodent models suggest that amphetamines and cocaine interact with different pathways to that affected by caffeine. Caffeine has a variety of pharmacological effects but its affinity for adenosine receptors is comparable with the levels expected to exist in the body after moderate caffeine intake, thus making adenosine receptor blockade the favoured mode of ergogenic action. However, alternative modes of action to account for the ergogenic properties of caffeine have been supported in the literature. Biochemical mechanisms that are consistent with more recent research findings, involving proteins such as DARPP-32 (dopamine and cAMP-regulated phosphoprotein), are helping to rationalize the molecular details of stimulant action in the central nervous system. PMID- 18384287 TI - Gene doping. AB - Gene doping is the misuse of gene therapy to enhance athletic performance. It has recently been recognised as a potential threat and subsequently been prohibited by the World Anti-Doping Agency. Despite concerns with safety and efficacy of gene therapy, the technology is progressing steadily. Many of the genes/proteins which are involved in determining key components of athletic performance have been identified. Naturally occurring mutations in humans as well as gene-transfer experiments in adult animals have shown that altered expression of these genes does indeed affect physical performance. For athletes, however, the gains in performance must be weighed against the health risks associated with the gene transfer process, whereas the detection of such practices will provide new challenges for the anti-doping authorities. PMID- 18384288 TI - Drug testing. AB - The analysis of sports samples for prohibited substances began in the 1960s and has developed since then using modern technologies close to the latest scientific discoveries. In this chapter the latest techniques and applications are described as well as the role of the World Anti-Doping Agency as the controlling body for the implementation of these tests. For small molecules, apart from the routine use of GC-MS, the newer techniques include the use of isotope ratio MS to detect testosterone and nandrolone administration and LC-MS/MS (liquid chromatography tandem MS) to detect diuretics. For large molecules, several applications of LC MS/MS are described as well as immunoprocedures for erythropoietin and human growth hormone. Finally, the latest method to detect homologous blood transfusion is briefly described. PMID- 18384289 TI - Conformation, catalytic site, and enzymatic mechanism of the PR10 allergen related enzyme norcoclaurine synthase. AB - The enzyme NCS [(S)-norcoclaurine synthase; EC 4.2.1.78] found in the common meadow rue, Thalictrum flavum, and other plant species, is involved in the biosynthesis of BIAs (benzylisoquinoline alkaloids). This group of plant secondary metabolites comprises pharmacologically-active compounds such as morphine and codeine. NCS catalyses the condensation of 4-HPAA (4 hydroxyphenylacetaldehyde) and dopamine to (S)-norcoclaurine, the common precursor of all plant BIAs. Although enzymatic properties of NCS and mechanistic aspects of the reaction have been studied in detail, no structural information on NCS was available so far. The enzyme shows significant sequence homology to members of the PR10 proteins (class 10 of pathogenesis-related proteins) such as the major birch pollen allergen Bet v 1. Our CD and NMR spectroscopic data indicated high similarity of the NCS and the Bet v 1 fold and allowed us to model NCS using Bet v 1 as a template. Virtually complete backbone assignment of the NCS sequence was used to study substrate binding by NMR titration experiments. Although binding of 4-HPAA seems to induce side-chain rearrangements in an extensive part of the protein, the putative distinct interaction site for dopamine could be clearly identified. The oligomerization state of NCS that reportedly plays an important role in enzyme functionality was determined to be concentration-dependent by SEC (size-exclusion chromatography) as well as NMR relaxation measurements, and the enzyme was found to be predominantly a monomer at the low micromolar concentrations used for activity assays. PMID- 18384290 TI - Identification of novel small-molecule histone deacetylase inhibitors by medium throughput screening using a fluorigenic assay. AB - HDACs (histone deacetylases) are considered to be among the most important enzymes that regulate gene expression in eukaryotic cells. In general, increased levels of histone acetylation are associated with increased transcriptional activity, whereas decreased levels are linked to repression of gene expression. HDACs associate with a number of cellular oncogenes and tumour-suppressor genes, leading to an aberrant recruitment of HDAC activity, which results in changes of gene expression, impaired differentiation and excessive proliferation of tumour cells. Therefore HDAC inhibitors are efficient anti-proliferative agents in both in vitro and in vivo pre-clinical models of cancer, making them promising anticancer therapeutics. In the present paper, we present the results of a medium throughput screening programme aiming at the identification of novel HDAC inhibitors using HDAH (HDAC-like amidohydrolase) from Bordetella or Alcaligenes strain FB188 as a model enzyme. Within a library of 3719 compounds, several new classes of HDAC inhibitor were identified. Among these hit compounds, there were also potent inhibitors of eukaryotic HDACs, as demonstrated by an increase in histone H4 acetylation, accompanied by a decrease in tumour cell metabolism in both SHEP neuroblastoma and T24 bladder carcinoma cells. In conclusion, screening of a compound library using FB188 HDAH as model enzyme identified several promising new lead structures for further development. PMID- 18384291 TI - Concerted action of two novel tRNA mtDNA point mutations in chronic progressive external ophthalmoplegia. AB - CPEO (chronic progressive external ophthalmoplegia) is a common mitochondrial disease phenotype in adults which is due to mtDNA (mitochondrial DNA) point mutations in a subset of patients. Attributing pathogenicity to novel tRNA mtDNA mutations still poses a challenge, particularly when several mtDNA sequence variants are present. In the present study we report a CPEO patient for whom sequencing of the mitochondrial genome revealed three novel tRNA mtDNA mutations: G5835A, del4315A, T1658C in tRNATyr, tRNAIle and tRNAVal genes. In skeletal muscle, the tRNAVal and tRNAIle mutations were homoplasmic, whereas the tRNATyr mutation was heteroplasmic. To address the pathogenic relevance, we performed two types of functional tests: (i) single skeletal muscle fibre analysis comparing G5835A mutation loads and biochemical phenotypes of corresponding fibres, and (ii) Northern-blot analyses of mitochondrial tRNATyr, tRNAIle and tRNAVal. We demonstrated that both the G5835A tRNATyr and del4315A tRNAIle mutation have serious functional consequences. Single-fibre analyses displayed a high threshold of the tRNATyr mutation load for biochemical phenotypic expression at the single cell level, indicating a rather mild pathogenic effect. In contrast, skeletal muscle tissue showed a severe decrease in respiratory-chain activities, a reduced overall COX (cytochrome c oxidase) staining intensity and abundant COX-negative fibres. Northern-blot analyses showed a dramatic reduction of tRNATyr and tRNAIle levels in muscle, with impaired charging of tRNAIle, whereas tRNAVal levels were only slightly decreased, with amino-acylation unaffected. Our findings suggest that the heteroplasmic tRNATyr and homoplasmic tRNAIle mutation act together, resulting in a concerted effect on the biochemical and histological phenotype. Thus homoplasmic mutations may influence the functional consequences of pathogenic heteroplasmic mtDNA mutations. PMID- 18384292 TI - Immune regulatory cells: circulating biomarker factories in cardiovascular disease. AB - TLRs (Toll-like receptors) are the first line of defence of the innate immune system. These receptors recognize not only exogenous but also endogenous ligands that are expressed following arterial injury and atherosclerotic disease. Expression of TLR2 and TLR4 is observed in macrophages and different vascular cells and is increased in atherosclerotic disease. It is suggested that TLR expression and functional responsiveness following ligation could serve as biomarkers for atherosclerotic disease progression. Recently published research papers support the concept that the TLRs are fast responders upon arterial injury. In the present issue of Clinical Science, Ishikawa and co-workers provide new evidence emerges that points to the potential prognostic properties of the TLRs. PMID- 18384338 TI - Hepatic tolerance of repeated intravenous paracetamol administration in neonates. AB - INTRODUCTION: An intravenous (i.v.) formulation of paracetamol is available, but reports on its hepatic tolerance in neonates are limited. We therefore assessed hepatic tolerance of i.v. paracetamol in neonates. METHODS: In a single centre retrospective study, clinical data and hepatic enzyme profiles (ALT, AST, gammaGT) were collected in neonates treated with i.v. paracetamol between January 1, 2006 and October 1, 2007. Hepatic enzyme profiles were retrieved from 2 days before until 2 days after i.v. paracetamol administration. Mann-Whitney U-test was used to compare hepatic enzymes before, during, and after i.v. treatment. Correlations (Spearman rank) of hepatic enzymes with duration of treatment during i.v. administration were investigated. RESULTS: In 189 cases, 2360 administrations {postmenstrual age 38 (range 30-55) weeks, postnatal age 5 (1 182) days} were documented and 1132 hepatic enzyme observations were available in 149/189 cases. There was no significant increase in ALT, AST, or gammaGT when pretreatment observations (n = 310) were compared with observations during (n = 649) or during with after (n = 173) treatment, nor was there a significant increase during administration. CONCLUSIONS: This retrospective study on hepatic tolerance provides evidence on safety aspects of i.v. paracetamol in neonates. Future studies should focus on dose-findings and pharmacodynamics of this formulation in neonates. PMID- 18384339 TI - The effect of a preoperative education programme on perioperative anxiety in children: an observational study. AB - The distress of children at the induction of anesthesia (DAI) is unpleasant for all involved and potentially harmful. Many strategies such as premedication or parental presence at induction have been described to minimize it. A preoperative education programme [the 'Saturday Morning Club' or (SMC)] has been in existence in our institution for a number of years and an observational study of children undergoing day case surgery was undertaken to assess the influence of attendance at the SMC on DAI. Ninety-four children aged between 2 and 16 years of age were included in the study; 21 attended the SMC and 73 did not. Patient anxiety using the modified Yale Preoperative Anxiety Scale was measured by blinded observers on the day ward, in the preoperative waiting room and at induction of anesthesia. Parental anxiety at the same locations was self reported using a visual analogue scale. Attendance at the SMC had a favorable effect on patient anxiety levels in all three locations but only reached statistical significance in the waiting room (P = 0.007, Mann-Whitney U-test). At present there is little evidence to support the use of preoperative education programmes in the UK and further studies are required to determine their benefit. PMID- 18384341 TI - Effects of continuous positive airway pressure and helium inhalation on thoracoabdominal asynchrony in an infant with post-extubation upper airway obstruction. PMID- 18384340 TI - Extremely low preanesthetic BIS values in two children with West syndrome and lissencephaly. PMID- 18384343 TI - Glycoprotein IIb/IIIa receptor inhibitors in 2008: do they still have a role? PMID- 18384344 TI - Intraarterial guidewire external compression: a simple technique for successful LIMA angiography through the right radial approach. AB - Specific techniques have been described for performing a left internal mammary artery (LIMA) angiography through the right radial approach. The unsuccessful LIMA angiography in anatomic situations, such as aorta dilatation or a far distal anatomical origin of the LIMA, among others, is a main problem not yet solved even with these techniques. In this report, we describe a simple and ready maneuver that can be helpful for a successful LIMA angiography in such situations. PMID- 18384347 TI - Molar incisor hypomineralization: a study of aetiological factors in a group of UK children. AB - OBJECTIVES: The objectives were to investigate the aetiology of molar incisor hypomineralization (MIH), and to discuss previously published studies. SUBJECTS AND METHODS: One hundred and nine children attending the Department of Paediatric Dentistry at the Royal London Hospital were included in the study: 57 children with MIH and 52 controls. Their mothers completed a medical history interview. RESULTS: No significant associations were found with MIH and delivery and birth complications, breastfeeding, immunization history, other illnesses and allergies, general anaesthetics, fluoride history, and trauma or abscesses affecting the primary predecessors. A family history of enamel defects was more commonly reported for MIH children, but the association was not statistically significant. However, MIH was significantly more common among those whose mothers had experienced problems during pregnancy (P = 0.025), those who had chickenpox between the ages of 3 and 3.99 (P = 0.047), and those for whom amoxycillin was the only antibiotic they had received (P = 0.028). CONCLUSIONS: The aetiology of MIH remains unclear, and this study questions whether it is because of a lone aetiological insult. This study recommends further research looking at the links with chickenpox occurring around the third year of life and amoxycillin. PMID- 18384348 TI - Clinical performance of a diode laser fluorescence device for the detection of occlusal caries in primary teeth. AB - OBJECTIVES: To correlate the DIAGNOdent readings (LF) with those of direct visual (DV) examination, indirect visual (IDV) examination, bitewing radiography (BWR), and pit and fissure opening (PFO) for the detection of occlusal caries in primary teeth; to determine the validity of this device using PFO as reference; and to evaluate its reliability. METHODS: Two calibrated operators examined 130 teeth with 405 examination sites using DV, IDV and radiographic examinations, and LF. Of the 405 sites, 155 were visually scored for caries after pit and fissure opening. RESULTS: Pit and fissure opening was found to significantly correlate with all methods (rhoLF = 0.48, rhoBWX = 0.48, rhoDV = 0.44, rhoIDV = 0.41). For enamel lesions, higher sensitivity (0.76) was found with DV, while higher specificity (0.88) with the LF. For lesions into dentin, however, higher sensitivity (0.78) was found with the LF, while higher specificity (0.98) with the BWR. The device's accuracy was found to be 0.61 for enamel lesions, while for lesions into dentin 0.70, and its reliability was excellent (ICC = 0.97). CONCLUSION: The LF device presented high reliability in the detection of occlusal caries in primary teeth and its performance was similar to DV and radiographic examinations. PMID- 18384349 TI - Salivary microbiota of HIV-positive children and its correlation with HIV status, oral diseases, and total secretory IgA. AB - AIM: This study aimed to investigate the prevalence and levels of salivary microorganisms in HIV-positive children, and their correlation to HIV status, oral lesions, and salivary IgA levels. DESIGN: Forty-two HIV-positive and 36 control children were clinically examined, had their saliva collected and processed for the microbiological analysis of 38 bacterial taxa by the checkerboard method, and salivary IgA quantification by ELISA. RESULTS: The majority of the species tested were more prevalent in control children than in the HIV group. Mean concentration of total salivary IgA was similar in both groups. High levels of Veillonella parvula were found in children with cheilitis and herpes. Tannerella forsythia, Eikenella Corrodens, and Propionibacterium acnes were prevalent in children with gingivitis, while Fusobacterium periodonticum, Streptococcus gordonii, and Streptococcus oralis were significantly more frequent in children with no oral lesions. Significant negative correlations between salivary IgA levels and Eubacterium nodatum and oral streptococci were observed (P < 0.05). CONCLUSION: HIV-seropositive children presented significantly lower prevalence and levels of several bacterial species in saliva; HIV-positive children are able to mount a mucosal immune response; HIV seropositive children under highly active antiretroviral therapy presented low prevalence of oral lesions. PMID- 18384350 TI - Assessing the proposed association between tooth agenesis and taurodontism in 975 paediatric subjects. AB - BACKGROUND: An association between tooth agenesis and taurodontism has been suggested. The identification of subpopulations with specific associated dental anomalies (subphenotype) would allow testing of the specific hypothesis that certain genetic factors contribute to the specific subphenotype. AIM: This work aims to assess a large cohort to verify if the association between tooth agenesis and taurodontism is present. DESIGN: Panoramic radiographs of 1002 patients were examined and 975 were used in this study. The presence of tooth agenesis and taurodontism was assessed in the study population. RESULTS: The frequency of tooth agenesis was 4.6% and the frequency of taurodontism was 1.6%. There were, however, no observations of concomitant tooth agenesis and taurodontism. CONCLUSIONS: Our data do not support the hypothesis that isolated tooth agenesis is associated with isolated taurodontism. PMID- 18384351 TI - Factors contributing to inter- and intra-annual variation in condition of cod Gadus morhua in the Barents Sea. AB - 1. Relative body condition (the quantity of stored energy) is an important tool in understanding demographic variation and the ability of a population to respond to environmental stressors, varying food availability and competition. 2. A high resolution database was used to examine causes of variation in the condition of north-east Arctic cod (Gadus morhua L.) for the period 1967-2004, over annual and monthly timescales. Community dynamics and climate variation were also tested as potential causes. 3. Temperature was shown to have a positive impact on condition at both inter- and intra-annual timescales. Inter-annually, temperature may affect stock distribution, in particular its overlap with the capelin stock. At shorter timescales it is likely that temperature directly affects the metabolism of the cod. 4. Intra-annually, the quantity of capelin in cod stomachs positively affected cod condition in the current and the preceding month for all lengths of cod. This indicated a time lag between a change in food consumption and a subsequent change in condition, or 'latency'. 5. Our study has shown that variation in temperature is a vital determinant of changes in condition, both at inter- and intra-annual timescales. Furthermore, the principle of latency has been demonstrated at the population level. Indirect effects of competition for energy-rich resources have been shown to have a negative effect on condition. This study supplements our knowledge of the implications for condition of changes in climate and in potential food resources. PMID- 18384353 TI - Re-evaluation of three Israeli families initially diagnosed with type 1 von Willebrand disease in light of the ISTH update on von Willebrand factor pathophysiology and classification. PMID- 18384352 TI - Recombinant factor VIIa to prevent surgical bleeding in factor XI deficiency. AB - Factor XI (FXI) deficiency is associated with bleeding after invasive procedures. Risks of human plasma-derived FXI replacement products include transfusion transmitted infection, thrombosis and fluid overload. This study was designed to test the hypothesis that recombinant factor VIIa (rFVIIa) is an effective haemostatic agent in patients with FXI deficiency undergoing surgery. Fourteen FXI deficient patients [five severely deficient (FXI:C <20 U dL(-1)) and nine partially deficient (FXI:C 20-70 U dL(-1)] received rFVIIa to prevent surgical bleeding during five major, four minor and six dental procedures. Minor surgical and dental procedures were covered with two doses of rFVIIa (90 microg kg(-1) i.v.), the first pre-operatively and the second 4 h postoperatively. Major surgery was covered with 90 microg kg(-1) i.v. two hourly for the first 24 h and four hourly for the second 24 h. Oral tranexamic acid was given for 7 days postoperatively. Effective haemostasis was observed in all cases and no alternative haemostatic agents or blood transfusions were required. Three adverse events were recorded; an acute cerebrovascular accident in a patient with a history of cardiovascular disease, an allergic reaction and local phlebitis. In this study, rFVIIa was an effective alternative to plasma-derived FXI replacement for the prevention of surgical bleeding in FXI deficient patients but rFVIIa may not be suitable for patients with pre-existing risk factors for thrombosis. PMID- 18384354 TI - Application of intron 9 and intron 25 dinucleotide repeats of the factor VIII gene for carrier diagnosis in haemophilia A. AB - We describe the usefulness of two dinucleotide repeats located in intron 9 and in intron 25 of the factor VIII gene for carrier diagnosis of haemophilia A. We analyzed 100 unrelated Spanish women and 34 women from haemophilia A (HA) families in whom known intragenic markers were unhelpful in determining their carrier status. The heterozygosity rate of intron 9 and intron 25 markers in the 100 control women was lower (0.28 and 0.38, respectively) than the values obtained with common markers routinely used in our laboratory. However, the application of intron 9 and intron 25 markers was effective in identifying the at risk X chromosome in 11 of 34 (32%) of the uninformative women from HA families. The combined use of these repeats with current markers may facilitate the identification of the X chromosome in HA families for application in carrier, prenatal and pre-implantation diagnoses. PMID- 18384355 TI - The impact of severe haemophilia on the social status and quality of life among Austrian haemophiliacs. AB - Although many studies of the impact of haemophilia on the quality of life were conducted, there is hardly any data on the social status of haemophiliacs. It was the aim of our study to obtain data on the social status of Austrian haemophiliacs and to compare these with an age- and sex-matched reference population. Furthermore, we collected data on the quality of life of haemophilia patients. We conducted a case-control study in two Austrian haemophilia centres with 53 patients (mean age 36.7 +/- 10.6 years) and 104 male controls (mean age 36.7 +/- 11.1 years). Socio-demographic data were collected using a standardized questionnaire and quality-of-life data using the SF-36. More patients (56.6%) than controls (37.5%) were married (P = 0.023), whereas more controls (17.3%) than patients (3.8%) had a partner with whom they were not married (P = 0.016). The percentage having children was equivalent in both groups (47% and 41% respectively), but controls had more children (mean number 1.5 in patients and 2.1 in controls, P < 0.007). A greater number of patients was unemployed (34% of patients, 9% of controls, P < 0.001) as well as retired (23% and 4% respectively; P < 0.001). Patients had worse scores regarding physical functioning, role physical, bodily pain and general health (P < 0.001), whereas vitality, social functioning, role-emotional and mental health were similar in both groups. Despite their disability, most of the Austrian haemophiliacs share a sound family environment. This suggests that they are highly capable of coping with their chronic disease and is indicated by good scores for role-emotional and mental health. PMID- 18384357 TI - Perceptions of the causes of childhood disability among Pakistani families living in the UK. AB - It has been well documented that South Asian families caring for a child with a disability experience discrimination and disadvantage in accessing health and social care services. This gives increasing cause for concern as the number of South Asian people with severe learning impairments in the UK continues to rise. Mainstream services are ill-equipped to provide individual services to parents who choose to define and address disability differently, and whose concerns and solutions may differ from the majority population, despite the fact that these parents possess many strengths and may present alternative ways of addressing their needs. This study aimed to provide a detailed account of the ways that Pakistani parents living in a northern city in the UK and caring for a child with a disability, account for and understand their child's disability. Sixteen in depth interviews were carried out between October 2002 and July 2003, with Pakistani parents and one grandparent of children with a disability. All final transcripts were in English, and a thematic analysis was undertaken. Results include a detailed account of parents' understanding of the causes of their child's disability. All parents made reference to theological explanations as to why they had a child with a disability; however, most parents also gave biomedical or other explanations. The relationship between theological and biomedical explanations was explored along with the consequences of the different beliefs for individuals and families. Parents also spoke vividly about the impact of negative and stigmatizing beliefs about causes of disability which they felt were prevalent in the Pakistani community, and they used a biomedical discourse to refute these ideas. The authors suggest that knowledge of individual and family beliefs, and awareness of the impact of these beliefs, can provide a valuable conceptual lens for health and social care practitioners to use when working in family-centred, culturally congruent ways with parents. PMID- 18384356 TI - T-cell regulation by CD46 and its relevance in multiple sclerosis. AB - CD46 is a complement regulatory molecule expressed on every cell type, except for erythrocytes. While initially described as a regulator of complement activity, it later became a 'magnet for pathogens', binding to several viruses and bacteria. More recently, an alternative role for such complement molecules has emerged: they do regulate T-cell immunity, affecting T-cell proliferation and differentiation. In particular, CD46 stimulation induces Tr1 cells, regulatory T cells characterized by massive production of interleukin-10 (IL-10), a potent anti-inflammatory cytokine. Hence, CD46 is likely to control inflammation. Indeed, data from CD46 transgenic mice highlight a role for CD46 in inflammation, with antagonist roles depending on the cytoplasmic tail being expressed. Furthermore, recent data have shown that CD46 is defective in multiple sclerosis, IL-10 production being severely impaired in these patients. This lack of IL-10 production probably participates in the inflammation observed in patients with multiple sclerosis. This review will summarize the data on CD46 and T cells, and how CD46 is likely involved in multiple sclerosis. PMID- 18384358 TI - Costs and health outcomes of intermediate care: results from five UK case study sites. AB - The objectives of this study were to explore the costs and outcomes associated with different types of intermediate care (IC) services, and also to examine the characteristics of patients receiving such services. Five UK case studies of 'whole systems' of IC were used, with data collected on a sample of consecutive IC episodes between January 2003 and January 2004. Statistical differences in costs and outcomes associated with different IC services and patient groups were explored. Factors associated with variation in IC episode outcomes (EuroQol EQ-5D and Barthel Index) were explored using an econometric framework. Data were available for 2253 episodes of IC. In terms of Department of Health criteria, a large proportion of patients (up to 47% of those for whom data were available) in this study were inappropriately admitted to IC services. As regards service function, compared to supported discharge, admission avoidance services were associated with both lower costs and greater health and functional gains. These gains appear to be driven, in part, by illness severity (more dependent patients tended to gain most benefit). In addition, these gains appear to be larger where the admission was appropriate. Our work suggests a need for the development and application of robust and reliable clinical criteria for admission to IC, and close co-operation between hospital and community service providers over selection of patients and targeting of IC and acute care services to meet defined clinical need. PMID- 18384359 TI - Research use and support needs, and research activity in social care: a cross sectional survey in two councils with social services responsibilities in the UK. AB - The purpose of this study was to investigate the level of research activity, research use, research interests and research skills in the social care workforce in two UK councils with social service responsibilities (CSSRs). A cross sectional survey was conducted of the social care workforce in two CSSRs (n = 1512) in 2005. The sample was identified in partnership with the councils, and included employees with professional qualifications (social workers and occupational therapists); staff who have a role to assess, plan and monitor care; service managers; commissioners of services; and those involved with social care policy, information management and training. The survey achieved a response rate of 24% (n = 368). The Internet was reported as an effective source of research information; conversely, research-based guidelines were reported to have a low impact on practice. Significant differences were found in research use, by work location, and postgraduate training. Most respondents saw research as useful for practice (69%), and wanted to collaborate in research (68%), but only 11% were planning to do research within the next 12 months. Having a master's degree was associated with a greater desire to lead or collaborate in research. A range of research training needs, and the preferred modes of delivery were identified. Support to increase research activity includes protected time and mentorship. The study concludes that a range of mechanisms to make research available for the social care workforce needs to be in place to support evidence-informed practice. Continual professional development to a postgraduate level supports the use and production of evidence in the social care workforce, and promotes the development of a research culture. The term research is used to include service user consultations, needs assessment and service evaluation. The findings highlight a relatively large body of the social care workforce willing to collaborate and conduct research. Councils and research support systems need to be developed to utilise this relatively untapped potential. PMID- 18384360 TI - Effect of eliminating behavioral health benefits for selected medicaid enrollees. AB - OBJECTIVE: To determine the extent to which the elimination of behavioral health benefits for selected beneficiaries of Oregon's Medicaid program affected general medical expenditures among enrollees using outpatient mental health and substance abuse treatment services. DATA SOURCE/STUDY SETTING: Twelve months of claims before and 12 months following a 2003 policy change, which included the elimination of the behavioral health benefit for selected Oregon Medicaid enrollees. STUDY DESIGN: We use a difference-in-differences approach to estimate the change in general medical expenditures following the 2003 policy change. We compare two methodological approaches: regression with propensity score weighting; and one-to-one covariate matching. PRINCIPAL FINDINGS: Enrollees who had accessed the substance abuse treatment benefit demonstrated substantial and statistically significant increases in expenditures. Individuals who accessed the outpatient mental health benefit demonstrated a decrease or no change in expenditures, depending on model specification. CONCLUSIONS: Elimination of the substance abuse benefit led to increased medical expenditures, although this offset was still smaller than the total cost of the benefit. In contrast, individuals who accessed the outpatient mental health benefit did not exhibit a similar increase, although these individuals did not include a portion of the Medicaid population with severe mental illnesses. PMID- 18384363 TI - Disentangling interactions between adaptive divergence and gene flow when ecology drives diversification. AB - Adaptive diversification is driven by selection in ecologically different environments. In absence of geographical barriers to dispersal, this adaptive divergence (AD) may be constrained by gene flow (GF). And yet the reverse may also be true, with AD constraining GF (i.e. 'ecological speciation'). Both of these causal effects have frequently been inferred from the presence of negative correlations between AD and GF in nature - yet the bi-directional causality warrants caution in such inferences. We discuss how the ability of correlative studies to infer causation might be improved through the simultaneous measurement of multiple ecological and evolutionary variables. On the one hand, inferences about the causal role of GF can be made by examining correlations between AD and the potential for dispersal. On the other hand, inferences about the causal role of AD can be made by examining correlations between GF and environmental differences. Experimental manipulations of dispersal and environmental differences are a particularly promising approach for inferring causation. At present, the best studies find strong evidence that GF constrains AD and some studies also find the reverse. Improvements in empirical approaches promise to eventually allow general inferences about the relative strength of different causal interactions during adaptive diversification. PMID- 18384362 TI - Multimethod evaluation of health policy change: an application to Medicaid managed care in a rural state. AB - OBJECTIVE: To answer questions about the impacts of Medicaid managed care (MMC) at the individual, organizational/community, and population levels of analysis. DATA SOURCES/STUDY SETTING: Multimethod approach to study MMC in New Mexico, a rural state with challenging access barriers. STUDY DESIGN: Individual level: surveys to assess barriers to care, access, utilization, and satisfaction. Organizational/community level: ethnography to determine changes experienced by safety net institutions and local communities. Population level: analysis of secondary databases to examine trends in preventable adverse sentinel events. DATA COLLECTION/EXTRACTION METHODS: SURVEY: multivariate statistical methods, including factor analysis and logistic regression. Ethnography: iterative coding and triangulation to assess documents, field observations, and in-depth interviews. Secondary databases: plots of sentinel events over time. PRINCIPAL FINDINGS: The survey component revealed no consistent changes after MMC, relatively favorable experiences for Medicaid patients, and persisting access barriers for the uninsured. In the ethnographic component, safety net institutions experienced increased workload and financial stress; mental health services declined sharply. Immunization rate, as an important sentinel event, deteriorated. CONCLUSIONS: MMC exerted greater effects on safety net providers than on individuals and did not address problems of the uninsured. A multimethod approach can facilitate evaluation of change in health policy. PMID- 18384361 TI - Racial/ethnic disparities in knowledge about risks and benefits of breast cancer treatment: does it matter where you go? AB - OBJECTIVE: To evaluate the association between provider characteristics and treatment location and racial/ethnic minority patients' knowledge of breast cancer treatment risks and benefits. DATA SOURCES/DATA COLLECTION: Survey responses and clinical data from breast cancer patients of Detroit and Los Angeles SEER registries were merged with surgeon survey responses (N=1,132 patients, 277 surgeons). STUDY DESIGN: Cross-sectional survey. Multivariable regression was used to identify associations between patient, surgeon, and treatment setting factors and accurate knowledge of the survival benefit and recurrence risk related to mastectomy and breast conserving surgery with radiation. PRINCIPAL FINDINGS: Half (51 percent) of respondents had survival knowledge, while close to half (47.6 percent) were uncertain regarding recurrence knowledge. Minority patients and those with lower education were less likely to have adequate survival knowledge and more likely to be uncertain regarding recurrence risk than their counterparts (p<.001). Neither surgeon characteristics nor treatment location attenuated racial/ethnic knowledge disparities. Patient physician communication was significantly (p<.001) associated with both types of knowledge, but did not influence racial/ethnic differences in knowledge. CONCLUSIONS: Interventions to improve patient understanding of the benefits and risks of breast cancer treatments are needed across surgeons and treatment setting, particularly for racial/ethnic minority women with breast cancer. PMID- 18384364 TI - Farysizyma gen. nov., an anamorphic genus in the Ustilaginales to accommodate three novel epiphytic basidiomycetous yeast species from America, Europe and Asia. AB - Among many isolates that resulted from four independent surveys of yeasts associated with plants in Brazil, the USA, Portugal and Taiwan, we have characterized eighteen basidiomycetous strains, two of which were conspecific with the type strain of Rhodotorula acheniorum, whereas the remaining sixteen isolates appeared not to correspond to any previously described species. Microsatellite-PCR fingerprinting with primers M13 and (GTG)5 confirmed that the latter strains formed three genetically distinct groups. Each group was considered to represent a distinct species based on nucleotide sequences of the D1/D2 domains of the 26S rRNA gene and the internal transcribed spacer (ITS) region. Phylogenetic analyses of sequence data placed the putative novel species in a clade with R. acheniorum and the dimorphic smut fungus Farysia chardoniana. A novel anamorphic genus, Farysizyma, is created to accommodate the three undescribed species, which were named Farysizyma itapuensis, Farysizyma setubalensis and Farysizyma taiwaniana. A new combination, Farysizyma acheniorum, is proposed for R. acheniorum, which may represent the yeast-phase anamorph of Farysia thuemenii. PMID- 18384365 TI - Detection of Borrelia burgdorferi sensu lato DNA by PCR in serum of patients with clinical symptoms of Lyme borreliosis. AB - Lyme borreliosis is a disease caused by spirochaetes belonging to the genospecies complex Borrelia burgdorferi sensu lato (s.l.) transmitted by Ixodes ticks. At present, serology remains the main diagnostic tool for laboratory diagnosis of Lyme borreliosis. Recently, the PCR technique has been applied for diagnosis of B. burgdorferi s.l., but, until now, a reliable, easy-to-perform and sensitive method has not been described. Here we present a new PCR-based method for the detection of both B. burgdorferi s.l. and Borrelia genospecies DNAs in serum samples collected from patients showing Lyme disease symptoms. Of 265 serum samples of patients included in this study, 7.5% were positive, 1.9% was borderline and 90.6% were negative for antibodies against B. burgdorferi by enzyme-linked immunosorbent assay and Western blotting. The B. burgdorferi s.l. 16S rRNA gene was detected by PCR in all serum-positive and in two borderline samples. None of the serum-negative samples nor serum samples collected from healthy subjects gave positive PCR reactions. Of PCR-positive serum samples, 50% gave a positive reaction for Borrelia afzelii, 18% for Borrelia garinii and 23% for two Borrelia species. Two samples (9%) were not identified to species level. The new protocol could be considered to be reliable as neither false-positive nor false-negative reactions were recorded, and to be sensitive as it detects DNA from one bacterial cell. PMID- 18384366 TI - Toll-like receptors and fungal infections: the role of TLR2, TLR4 and MyD88 in paracoccidioidomycosis. AB - The aim of this minireview is to present a concise view of the most important pattern recognition receptors used by the innate immune system to sense and control pathogen growth into host tissues. A brief review of the role of Toll like receptors (TLRs) in fungal infections followed by some recent results on the function of TLR4, TLR2 and the MyD88 adaptor molecule in the pathogenesis of paracoccidioidomycosis are presented. PMID- 18384367 TI - Extraintestinal pathogenic isolates of Escherichia coli do not possess active IgA1, IgA2, sIgA or IgG proteases. AB - Infections outside of the intestinal tract due to pathogenic strains of Escherichia coli result in significant morbidity, mortality and increased healthcare costs. The ability of these strains to cause both mucosal and systemic infections, as well as recurrent infections due to the same (homologous) strain suggests the hypothesis that strains of E. coli that cause infection outside of the intestinal tract possess proteases that are capable of cleaving IgA1, IgA2, sIgA or IgG. To test this hypothesis the ability of eight E. coli strains, isolated from sites outside of the urinary tract and 14 homologous and 11 heterologous strains of E. coli that were isolated from women with recurrent UTI, to cleave IgA1, IgA2, sIgA or IgG was evaluated. Our experimental design allowed for detection of cell-associated and secreted immunoglobulin proteases in both log and stationary phase. Surprisingly, none of these 33 human clinical isolates when grown in iron depleted Luria-Bertani medium or human urine were able to degrade the immunoglobulins assessed. Despite previous studies suggesting otherwise, the findings from this study support the concept that strains of E. coli that cause infection outside of the intestinal tract do not possess proteases that cleave the human immunoglobulins IgA1, IgA2, sIgA or IgG. PMID- 18384368 TI - The cross-section trichometer: a new device for measuring hair quantity, hair loss, and hair growth. AB - BACKGROUND: Office physicians are unable to measure hair quantity, hair loss, and hair growth in a simple and meaningful manner. One solution is to measure the cross-sectional area of a bundle of hair that is growing within a premeasured cross-section of scalp. OBJECTIVE: The objective was to design a mechanical device that precisely measures the cross-sectional area of a bundle of hair and design a device that can precisely delineate an area of scalp. It was assumed that density and diameter changes are evidenced by changes in the bundle cross sectional area and that growth and loss are the result of density and diameter changes. These assumptions were confirmed using various sized bundles of known diameter non-hair filaments. MATERIALS AND METHODS: Bundles of hair and surgical silk fibers were tested using a mechanical device that compressed the bundle and measured its cross-sectional area. Balding patients were categorized according to their observed severity of the loss. Bundles of their uncut hair from 4-cm(2) scalp sites were measured and the values were compared to the patient's category of hair loss severity. RESULTS: In patients with balding, there was a direct correlation between the bundle's cross-sectional area and the observed severity of the loss. The cross-sectional area was expressed as square millimeters of hair per square centimeter of skin x 100 (mm(2)/cm(2)x 100) and named the trichometric index (TI). Using surgical silk fibers, there was a direct correlation between the bundle's cross-sectional area and the number of filaments, the diameter of the filaments, and the dry weight of the filament bundle. Using aggregates of cut human hair, there was a direct correlation between the cross-sectional area and the dry weight of the bundle. CONCLUSION: This prototype device shows promise as a diagnostic instrument for measuring changes in hair quantity (mass), hair diameter, and hair density, as evidenced by preliminary studies using silk sutures, cut human hair, and patients with various degrees of balding. Formal clinical studies are needed. Although the device itself showed a high degree of precision, the accuracy and reproducibility of the measurements can be compromised if the sampling method is not carefully performed using magnification. The device is intended for use on uncut hair that is more than 1 inch in length. PMID- 18384370 TI - Embryologic fusion planes: a plea for more precise analysis. PMID- 18384371 TI - Granuloma annulare as a complication of mesotherapy: a case report. PMID- 18384372 TI - Facial cutaneous ulcers following mesotherapy. PMID- 18384373 TI - Penile preservation surgery in a case of extramammary Paget's disease involving the glans penis and distal urethra. PMID- 18384374 TI - Subcutaneous forehead nodules: attention to the button osteoma and frontalis associated lipoma. PMID- 18384375 TI - Sp1-like sequences mediate human caspase-3 promoter activation by p73 and cisplatin. AB - Caspase-3 is a cysteine protease that plays a central role in the execution of apoptosis induced by a wide variety of stimuli. However, little is known about the mechanisms involved in the regulation of caspase-3 gene transcription. This study was carried out to characterize the human caspase-3 promoter and to understand the mechanisms involved in the induction of caspase-3 gene expression in response to the anticancer drug cisplatin and p73. Caspase-3 gene expression was induced by treatment of cells with cisplatin, which also induced p73 protein in HeLa and K562 cells. The human caspase-3 promoter was cloned and characterized. p73beta strongly activated the caspase-3 promoter, whereas p73alpha showed less activation. Cisplatin treatment increased caspase-3 promoter activity. Basal and cisplatin-induced promoter activity was inhibited by the p73 inhibitor p73DD. Deletion analysis defined a minimal promoter of 120 base pairs, which showed good basal and p73beta-induced activity. The examination of the minimal promoter sequence showed several putative Sp1 sites, but no p53/p73 site. The caspase-3 promoter was activated by Sp1 in Sp1-deficient Drosophila SL-2 cells. Sp1-induced promoter activity was further enhanced by p73beta in SL-2 cells. Mutation of Sp1 sites in the minimal promoter resulted in a loss of basal and p73-induced promoter activity. These results show that caspase-3 gene transcription is induced by cisplatin, which is mediated partly by p73. We have identified p73 and Sp1 as activators of the caspase-3 promoter. Sp1-like sequences in the minimal promoter not only sustain basal promoter activity, but also mediate p73-induced activation of the promoter. PMID- 18384376 TI - Analysis of the regulatory motifs in eukaryotic initiation factor 4E-binding protein 1. AB - Mammalian target of rapamycin complex 1 (mTORC1) phosphorylates proteins such as eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) and the S6 kinases. These substrates contain short sequences, termed TOR signalling (TOS) motifs, which interact with the mTORC1 component raptor. Phosphorylation of 4E-BP1 requires an additional feature, termed the RAIP motif (Arg-Ala-Ile-Pro). We have analysed the interaction of 4E-BP1 with raptor and the amino acid residues required for functional RAIP and TOS motifs, as assessed by raptor binding and the phosphorylation of 4E-BP1 in human cells. Binding of 4E-BP1 to raptor strongly depends on an intact TOS motif, but the RAIP motif and additional C terminal features of 4E-BP1 also contribute to this interaction. Mutational analysis of 4E-BP1 reveals that isoleucine is a key feature of the RAIP motif, that proline is also very important and that there is greater tolerance for substitution of the first two residues. Within the TOS motif, the first position (phenylalanine in the known motifs) is most critical, whereas a wider range of residues function in other positions (although an uncharged aliphatic residue is preferred at position three). These data provide important information on the structural requirements for efficient signalling downstream of mTORC1. PMID- 18384377 TI - Improvement of a monopartite ecdysone receptor gene switch and demonstration of its utility in regulation of transgene expression in plants. AB - In plants, regulation of transgene expression is typically accomplished through the use of inducible promoter systems. The ecdysone receptor (EcR) gene switch is one of the best inducible systems available to regulate transgene expression in plants. However, the monopartite EcR gene switches developed to date require micromolar concentrations of ligand for activation. We tested several EcR mutants that were generated by changing one or two amino acid residues in the highly flexible ligand-binding domain of Choristoneura fumiferana EcR (CfEcR). Based on the transient expression assays, we selected a double mutant, V395I + Y415E (VY), of CfEcR (CfEcR(VY)) for further testing in stable transformation experiments. The CfEcR(VY) mutant only slightly improved the induction characteristics of the two-hybrid gene switch, whereas the CfEcR(VY) mutant significantly improved the induction characteristics of the monopartite gene switch (VGCfE(VY)). The ligand sensitivity of the VGCfE(VY) switch was improved by 125-15 625-fold in different transgenic lines analyzed, compared to the VGCfE(Wt) switch. The utility of the VGCfE(VY) switch was tested by regulating the expression of an Arabidopsis zinc finger protein gene (AtZFP11) in both tobacco and Arabidopsis plants. These data showed that the VGCfE(VY) switch efficiently regulated the expression of AtZFP11 and that the phenotype of AtZFP11 could be induced by the application of ligand. In addition, the affected plants recovered after withdrawal of the ligand, demonstrating the utility of this gene switch in regulating the expression of critical transgenes in plants. PMID- 18384378 TI - Structural studies of nucleoside analog and feedback inhibitor binding to Drosophila melanogaster multisubstrate deoxyribonucleoside kinase. AB - The Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (dNK; EC 2.7.1.145) has a high turnover rate and a wide substrate range that makes it a very good candidate for gene therapy. This concept is based on introducing a suicide gene into malignant cells in order to activate a prodrug that eventually may kill the cell. To be able to optimize the function of dNK, it is vital to have structural information of dNK complexes. In this study we present crystal structures of dNK complexed with four different nucleoside analogs (floxuridine, brivudine, zidovudine and zalcitabine) and relate them to the binding of substrate and feedback inhibitors. dCTP and dGTP bind with the base in the substrate site, similarly to the binding of the feedback inhibitor dTTP. All nucleoside analogs investigated bound in a manner similar to that of the pyrimidine substrates, with many interactions in common. In contrast, the base of dGTP adopted a syn-conformation to adapt to the available space of the active site. PMID- 18384379 TI - Treatment with small interfering RNA affects the microRNA pathway and causes unspecific defects in zebrafish embryos. AB - MicroRNAs (miRNAs) are generated from primary transcripts through sequential processing by two RNase III enzymes, Drosha and Dicer, in association with other proteins. This maturation is essential for their function as post-transcriptional regulators. Notably, Dicer is also a component of RNA-induced silencing complexes, which incorporate either miRNA or small interfering RNA (siRNA) as guides to target specific mRNAs. In zebrafish, processed miRNAs belonging to the miR-430 family have previously been shown to promote deadenylation and degradation of maternal mRNAs during early embryogenesis. We show that injection of one-cell-stage zebrafish embryos with siRNA causes a significant reduction in the endogenous levels of processed miR-430 and other miRNAs, leading to unspecific developmental defects. Coinjection of siRNA with preprocessed miR-430 efficiently rescued development. This indicates that the abnormalities generally observed in siRNA-treated zebrafish embryos could be due to inhibition of miR-430 processing and/or activity. Our results also suggest that the miRNA pathway in mammals, under some experimental or therapeutic conditions, may be affected by siRNA. PMID- 18384380 TI - Biologically active, non membrane-anchored precursors: an overview. AB - Peptides function as chemical signals between cells of multicellular organisms via specific receptors on target cells. Many hormones, neuromodulators and growth factors are peptides. Peptide hormones and other biologically active peptides are synthesized as higher molecular weight precursor proteins (pro-hormones), which must undergo post-translational modification to yield the bioactive peptide(s). In many instances, more than one biologically active peptide is generated from one and the same precursor. In most cases, these precursors are biologically inert and their existence is confined to the membrane-enclosed subcellular compartments where processing of the pro-hormones takes place. A class of growth factors that derive from membrane-anchored precursors which themselves are biologically active constitute an exception to this model. The list of the membrane-anchored biologically active precursors has been the subject of specialized reviews. The present review focuses on precursors other than membrane anchored precursors, which were found to be biologically active and which often display different biological activities, and may mediate their effects via receptors independent from those of their generated peptides. PMID- 18384381 TI - Changes in acetylcholine receptor function induce shifts in muscle fiber type composition. AB - AChRepsilon(-/-) mice lack epsilon-subunits of the acetylcholine receptor and thus fail to express adult-type receptors. The expression of fetal-type receptors throughout postnatal life alters postsynaptic signal transduction and causes a fast-to-slow fiber type transition, both in slow-twitch soleus muscle and in fast twitch extensor digitorum longus muscle. In comparison to wild-type muscle, the proportion of type 1 slow fibers is significantly increased (6%), whereas the proportion of fast fibers is reduced (in soleus, type 2A by 12%, and in extensor digitorum longus, type 2B/2D by 10%). The increased levels of troponin I(slow) transcripts clearly support a fast-to-slow fiber type transition. Shifts of protein and transcript levels are not restricted to 'myogenic' genes but also affect 'synaptogenic' genes. Clear increases are observed for acetylcholine receptor alpha-subunits and the postsynaptically located utrophin. Although the fast-to-slow fiber type transition appears to occur in a coordinated manner in both muscle types, muscle-specific differences are retained. Most prominently, the differential expression level of the synaptic regulator MuSK is significantly lower in extensor digitorum muscle than in soleus muscle. The results show a new quality in muscle plasticity, in that changes in the functional properties of endplate receptors modulate the contractile properties of skeletal muscles. Muscle thus represents a self-matching system that adjusts contractile properties and synaptic function to variable functional demands. PMID- 18384382 TI - Chemical approaches to mapping the function of post-translational modifications. AB - Strategies for the chemical construction of synthetic proteins with precisely positioned post-translational modifications or their mimics offer a powerful method for dissecting the complexity of functional protein alteration and the associated complexity of proteomes. PMID- 18384383 TI - Branched N-glycans regulate the biological functions of integrins and cadherins. AB - Glycosylation is one of the most common post-translational modifications, and approximately 50% of all proteins are presumed to be glycosylated in eukaryotes. Branched N-glycans, such as bisecting GlcNAc, beta-1,6-GlcNAc and core fucose (alpha-1,6-fucose), are enzymatic products of N-acetylglucosaminyltransferase III, N-acetylglucosaminyltransferase V and alpha-1,6-fucosyltransferase, respectively. These branched structures are highly associated with various biological functions of cell adhesion molecules, including cell adhesion and cancer metastasis. E-cadherin and integrins, bearing N-glycans, are representative adhesion molecules. Typically, both are glycosylated by N acetylglucosaminyltransferase III, which inhibits cell migration. In contrast, integrins glycosylated by N-acetylglucosaminyltransferase V promote cell migration. Core fucosylation is essential for integrin-mediated cell migration and signal transduction. Collectively, N-glycans on adhesion molecules, especially those on E-cadherin and integrins, play key roles in cell-cell and cell-extracellular matrix interactions, thereby affecting cancer metastasis. PMID- 18384384 TI - Arginine-induced conformational change in the c-ring/a-subunit interface of ATP synthase. AB - The rotational mechanism of ATP synthases requires a unique interface between the stator a subunit and the rotating c-ring to accommodate stability and smooth rotation simultaneously. The recently published c-ring crystal structure of the ATP synthase of Ilyobacter tartaricus represents the conformation in the absence of subunit a. However, in order to understand the dynamic structural processes during ion translocation, studies in the presence of subunit a are required. Here, by intersubunit Cys-Cys cross-linking, the relative topography of the interacting helical faces of subunits a and c from the I. tartaricus ATP synthase has been mapped. According to these data, the essential stator arginine (aR226) is located between the c-ring binding pocket and the cytoplasm. Furthermore, the spatially vicinal residues cT67C and cG68C in the isolated c-ring structure yielded largely asymmetric cross-linking products with aN230C of subunit a, suggesting a small, but significant conformational change of binding-site residues upon contact with subunit a. The conformational change was dependent on the positive charge of the stator arginine or the aR226H substitution. Energy minimization calculations revealed possible modes for the interaction between the stator arginine and the c-ring. These biochemical results and structural restraints support a model in which the stator arginine operates as a pendulum, moving in and out of the binding pocket as the c-ring rotates along the interface with subunit a. This mechanism allows efficient interaction between subunit a and the c-ring and simultaneously allows almost frictionless movement against each other. PMID- 18384385 TI - Two L-amino acid oxidase isoenzymes from Russell's viper (Daboia russelli russelli) venom with different mechanisms of inhibition by substrate analogs. AB - Two isoforms, L(1) and L(2), of L-amino acid oxidase have been isolated from Russell's viper venom by Sephadex G-100 gel filtration followed by CM-Sephadex C 50 ion exchange chromatography. The enzymes, with different isoelectric points, are monomers of 60-63 kDa as observed from size exclusion HPLC and SDS/PAGE. Partial N-terminal amino acid sequencing of L(1) and L(2) showed significant homology with other snake venom L-amino acid oxidases. Both the enzymes exhibit marked substrate preference for hydrophobic amino acids, maximum catalytic efficiency being observed with L-Phe. Inhibition of L(1) and L(2) by the substrate analogs N-acetyltryptophan and N-acetyl-L-tryptophan amide has been followed. The initial uncompetitive inhibition of L(1) followed by mixed inhibition at higher concentrations suggested the existence of two different inhibitor-binding sites distinct from the substrate-binding site. In the case of L(2), initial linear competitive inhibition followed by mixed inhibition suggested the existence of two nonoverlapping inhibitor-binding sites, one of which is the substrate-binding site. An inhibition kinetic study with O aminobenzoic acid, a mimicking substrate with amino, carboxylate and hydrophobic parts, indicated the presence of three and two binding sites in L(1) and L(2), respectively, including one at the substrate-binding site. An inhibitor cross competition kinetic study indicated mutually excluding binding between N acetyltryptophan, N-acetyl-L-tryptophan amide and O-aminobenzoic acid in both the isoforms, except at the substrate-binding site of L(1). Binding of substrate analogs with different electrostatic and hydrophobic properties provides useful insights into the environment of the catalytic sites. Furthermore, it predicts the minimum structural requirement for a ligand to enter and anchor at the respective functional sites of LAAO that may facilitate the design of suicidal inhibitors. PMID- 18384386 TI - Autism and diagnostic substitution: evidence from a study of adults with a history of developmental language disorder. AB - Rates of diagnosis of autism have risen since 1980, raising the question of whether some children who previously had other diagnoses are now being diagnosed with autism. We applied contemporary diagnostic criteria for autism to adults with a history of developmental language disorder, to discover whether diagnostic substitution has taken place. A total of 38 adults (aged 15-31y; 31 males, seven females) who had participated in studies of developmental language disorder during childhood were given the Autism Diagnostic Observation Schedule--Generic. Their parents completed the Autism Diagnostic Interview--Revised, which relies largely on symptoms present at age 4 to 5 years to diagnose autism. Eight individuals met criteria for autism on both instruments, and a further four met criteria for milder forms of autistic spectrum disorder. Most individuals with autism had been identified with pragmatic impairments in childhood. Some children who would nowadays be diagnosed unambiguously with autistic disorder had been diagnosed with developmental language disorder in the past. This finding has implications for our understanding of the epidemiology of autism. PMID- 18384387 TI - Is the modified Tardieu scale in semi-standing position better associated with knee extension and hamstring activity in terminal swing than the supine Tardieu? AB - The aim of this study was to investigate whether the modified Tardieu scale (MTS) in a semi-standing position, used for the assessment of hamstrings spasticity, was better associated with knee extension and hamstrings activity in terminal swing than the MTS in a supine position in children with cerebral palsy (CP). Seven children diagnosed with spastic CP (Gross Motor Function Classification System Levels I-II) and seven healthy comparison children participated in the study. An instrumented MTS in supine and semi-standing position and an instrumented gait assessment were conducted. Results showed that spasticity related outcomes of the semi-standing MTS do not show better associations with terminal swing characteristics of gait than the same outcomes of the supine MTS in children with spastic CP. Only the passive restricted knee angle from the supine MTS was strongly associated with the maximum knee extension during gait (r(s)=0.99; p <0.001), suggesting that hamstrings length is more important for terminal swing behaviour than hamstrings spasticity. PMID- 18384388 TI - Spider silk fibres in artificial nerve constructs promote peripheral nerve regeneration. AB - OBJECTIVE: In our study, we describe the use of spider silk fibres as a new material in nerve tissue engineering, in a 20-mm sciatic nerve defect in rats. MATERIALS AND METHODS: We compared isogenic nerve grafts to vein grafts with spider silk fibres, either alone or supplemented with Schwann cells, or Schwann cells and matrigel. Controls, consisting of veins and matrigel, were transplanted. After 6 months, regeneration was evaluated for clinical outcome, as well as for histological and morphometrical performance. RESULTS: Nerve regeneration was achieved with isogenic nerve grafts as well as with all constructs, but not in the control group. Effective regeneration by isogenic nerve grafts and grafts containing spider silk was corroborated by diminished degeneration of the gastrocnemius muscle and by good histological evaluation results. Nerves stained for S-100 and neurofilament indicated existence of Schwann cells and axonal re-growth. Axons were aligned regularly and had a healthy appearance on ultrastructural examination. Interestingly, in contrast to recently published studies, we found that bridging an extensive gap by cell-free constructs based on vein and spider silk was highly effective in nerve regeneration. CONCLUSION: We conclude that spider silk is a viable guiding material for Schwann cell migration and proliferation as well as for axonal re growth in a long-distance model for peripheral nerve regeneration. PMID- 18384389 TI - Surfactive and antibacterial activity of cetylpyridinium chloride formulations in vitro and in vivo. AB - AIM: To compare effects of three cetylpyridinium chloride (CPC) formulations with and without alcohol and Tween80 on physico-chemical properties of salivary pellicles, bacterial detachment in vitro and bacterial killing in vivo. MATERIAL AND METHODS: Adsorption of CPC to salivary pellicles in vitro was studied using X ray photoelectron spectroscopy and water contact angle measurements. Adhesion and detachment of a co-adhering bacterial pair was determined in vitro using a flow chamber. Killing was evaluated after live/dead staining after acute single use in vivo on 24- and 72-h-old plaques after 2-week continuous use. RESULTS: The most pronounced effects on pellicle surface chemistry and hydrophobicity were observed after treatment with the alcohol-free formulation, while the pellicle thickness was not affected by any of the formulations. All CPC formulations detached up to 33% of the co-adhering pair from pellicle surfaces. Bacterial aggregate sizes during de novo deposition were enhanced after treatment with the alcohol-free formulation. Immediate and sustained killing in 24 and 72 h plaques after in vivo, acute single use as well as after 2-week continuous use were highest for the alcohol-free formulation. CONCLUSIONS: CPC bioavailability in a formulation without alcohol and Tween80 could be demonstrated through measures of pellicle surface properties and bacterial interactions in vitro as well as bacteriocidal actions on oral biofilms in vivo. PMID- 18384390 TI - TNF-alpha TGF-beta2 and IL-1beta levels in gingival and peri-implant crevicular fluid before and after de novo plaque accumulation. AB - AIMS: The aim of this split-mouth study was to investigate levels of tumour necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta2) and interleukin-1 beta (IL-1beta) in gingival crevicular fluid (GCF) and peri implant crevicular fluid (PICF) after a 21-day-period of de novo plaque accumulation in the same patient. MATERIAL AND METHODS: In 25 patients, samples of GCF and PICF were collected in the sulcus of the tooth and of the implant after professional hygiene. After the no-hygiene phase (21 days), second samples of GCF and PICF were taken. Third samples were collected after 69 days of re establishment oral hygiene techniques. The crevicular fluids were used to determine the volume and the levels of TNF-alpha, TGF-beta2 and IL-1beta. RESULTS: The volume of the crevicular fluids increased significantly after 21 days of plaque accumulation around teeth and implants and decreased significantly by 69 days. TNF-alpha and TGF-beta2 did not change significantly among the three different samples. A significant increase of IL-1beta was observed after plaque accumulation around the teeth GCF, whereas in the PICF the increase was not statistically significant. CONCLUSIONS: These data suggest that increased volumes of GCF and PICF could be useful markers of early inflammation in gingival and peri-implant tissues. In the presence of de novo plaque, implants showed lower, and nearly significant, levels of IL-1beta compared with teeth. PMID- 18384391 TI - Local minocycline as an adjunct to surgical therapy in moderate to severe, chronic periodontitis. AB - OBJECTIVE: The aim of the study was to evaluate the effects of minocycline microspheres on periodontal probing depth reduction when used in combination with surgery in adults with moderate to severe, chronic periodontitis. MATERIAL AND METHODS: Sixty patients with a minimum of one non-molar periodontal site > or =6 mm in two oral quadrants received either local minocycline microspheres at baseline, immediately following each of two surgical therapies (Weeks 2 and 3), and at Week 5 or surgery alone. RESULTS: The mean probing depth reduction at Week 25 at sites > or =5 mm at baseline was 2.51 mm in the test group and 2.18 mm in the control group. Smokers in the test group had a significantly greater probing depth reduction (2.30 mm) than smokers in the control group (2.05 mm). The number of sites with probing depth reductions of > or =2 and > or =3 mm were significantly higher in the test group than in the control group. CONCLUSION: Applications of local minocycline as an adjunct to surgery in adults with moderate to severe, chronic periodontitis were associated with statistically significant greater reductions in probing depth than surgery alone. PMID- 18384392 TI - Use of real-time PCR to detect human papillomavirus-16 viral loads in vaginal and urine self-sampled specimens. AB - Increasing the accuracy of self-sampling methods to detect oncogenic human papillomavirus (HPV) infection would contribute to the wider application of these approaches. In this study, 120 women were tested for HPV-16 by conventional and quantitative real-time PCR (QRT-PCR) in cervical and self-sampled vaginal and urine specimens. QRT-PCR had a higher detection rate, and the HPV viral load in all three sampling sites correlated with the severity of disease, as determined by histology. The vaginal and urine viral loads correlated with HPV-16 positivity according to both conventional and QRT-PCR, and were proportional to the cervical viral load. PMID- 18384393 TI - A rapid flow cytometric technique for the detection of platelet-monocyte complexes, activated platelets and platelet-derived microparticles. AB - Platelet activation occurs in a variety of clinical situations in which it directly contributes to the pathology. This study reports a simple flow cytometric assay for platelet activation which measures platelet-derived microparticles, activated platelets and platelet-monocyte complexes. Pre- and post analytical conditions were investigated and optimized and a normal range established on 20 healthy controls. Twenty patients pre- and post percutaneous coronary intervention (PCI) were tested with the technique. Soluble activation markers sCD40 ligand and sP-selectin and plasma phospholipid levels were measured in both groups. There was a significant increase in activated platelets and platelet-monocyte complexes between normal and pre-PCI (P = 0.005 and 0.0275, respectively) suggesting an activated state. There was a significant fall in activated platelets post-PCI (P = 0.0027) which was mirrored by a fall in soluble CD40 ligand, soluble P-selectin and plasma phospholipid levels (P = 0.0066, <0.0001 and 0.0032, respectively) consistent with antiplatelet therapy administered during the process. This is a reliable and rapid method for the assessment of ex vivo platelet activation which may be an aid in diagnosis and help guide therapy for patients with thrombotic disease. PMID- 18384394 TI - Levels of glycated haemoglobin A--HbA1c in nondiabetic carriers of haemoglobin O Arab. PMID- 18384395 TI - Indicators of iron status in elite soccer players during the sports season. AB - The main aim of the study was to investigate the changes of hematologic status in elite soccer players throughout a competitive season. Study was conducted with 35 male professional soccer players and the measurements were collected at the start of the conditioning period, at the start of the season, in the mid-season and at the end of the season. Blood was drawn from an antecubital vein for a complete blood count, serum iron, and transferrin and ferritin levels. We found significantly higher hematocrit at preseason assessment as compared with other sampling periods (P < 0.05). No other differences were found between any of the hematologic variables during the study. The lowest hemoglobin level, mean corpuscular volume (MCV), serum iron, ferritin and transferrin encountered in the study were Hb of 11.2 g/dl, MCV of 77.9 fl, serum iron of 34 microg/dl, ferritin of 15.1 microg/l, transferrin of 224 microg/dl in a 20-year-old soccer player at the start of the season. The variability of the iron status indicators in elite soccer players seems to be stable and poorly related to training phase during sports season. PMID- 18384396 TI - Promoter methylation of P15(INK4B) gene is possibly associated with parvovirus B19 infection in adult acute leukemias. AB - In this study, we examined the P15(INK4B) gene promoter methylation in patients with myelodysplastic syndrome and acute leukemia and its possible relationship with parvovirus B19 and Epstein-Barr virus infections. P15(INK4B) methylation frequency was significantly higher in acute leukemia patients than in that of non malignant patients (P < 0.05). When the patients with myelodysplastic syndrome were included, no significant difference was found between these groups regarding the methylation status. The possible correlation between P15(INK4B) promoter methylation and parvovirus B19 infection was observed in adult acute leukemia patients (P < 0.05). However, no similar relationship in EBV-infected patients was observed. To the best of our knowledge, this is the first report showing the possible association between P15(INK4B) promoter methylation and parvovirus B19 infection in acute leukemia. PMID- 18384397 TI - A retrospective analysis of early and delayed loading of full-arch mandibular prostheses using three different implant systems: clinical results with up to 5 years of loading. AB - BACKGROUND: Early loading of implant-supported prostheses in the edentulous mandible is widely accepted, but do the clinical results replicate those of delayed loading? PURPOSE: The aim of this study was to evaluate clinical outcome and patient satisfaction with early or delayed loading in patients treated with fixed prostheses, using three different implant systems. MATERIALS AND METHODS: One hundred and nine consecutively treated patients received 490 implants supporting fixed prostheses; 82 patients with Branemark System implants (Nobel Biocare AB, Goteborg, Sweden), 16 with Astra Tech implants (Astra Tech AB Dental Implant system, Molndal, Sweden), and 11 with ITI MonoType implants (ITI Dental Implant System, Institute Straumann AG, Waldenburg, Switzerland). Prostheses were placed within 2 to 3 weeks in 55 patients; 54 patients underwent a two-stage procedure. Data were collected from patient records and radiographs; 83 patients attended a clinical examination and received a questionnaire. RESULTS: All patients had fixed prostheses at follow-up with a mean observation time of 3.5 years. Cumulative survival rates (CSRs) were 92.5% of prostheses and 94.4% of implants for early loading, and 98.0 and 97.9% for delayed loading. The mean radiographic bone loss after the first year was small, and at 5 years less than 0.2 mm for both groups. With early loading, significantly more prostheses (p < .05) needed adjustment or replacement. CONCLUSION: Statistically significantly more prostheses needed adjustment or replacement in the early group. The present study suggested lower CSRs for prostheses and implants in the early loading group after 5 years; the difference was not statistically significant. Larger study samples are needed to verify statistically small differences between treatment techniques. PMID- 18384398 TI - Radiographic analyses of "advanced" marginal bone loss around Branemark dental implants. AB - BACKGROUND: Dental implant failures have a multifactorial background; dependency within patient/jaw exists. Failures caused by bone loss are rare. Lately, advanced bone loss around implants has been discussed. PURPOSES: Our aim was to study advanced bone level changes (>or=2 mm) regarding "clustering effect," prediction, and dependency. Further, we also aimed to study if the number of radiographs/radiographic examinations could be reduced. MATERIALS AND METHODS: Six hundred and forty patients (3,462 Branemark implants) with radiographic follow-ups >or=5 years were included, whereas patients with overdentures and augmentation procedures were excluded. RESULTS: Progression rate for implants with advanced bone loss was largest during the first year; thereafter, slow. A cluster effect was found with more advanced bone loss in few patients. Position was important for lower jaw implants with larger bone loss for implants placed close to midline. Age, jaw type, and implant placement were identified as predictors. The longer the follow-ups, the more bone loss around a randomly selected and examined implant, and the more implants per patient, the higher the risk for bone loss >or=2 mm around any other implant. Still, it seems safe to exclude radiographic follow-ups during the first 5 years. Dependency within the patient was found, hence the "one-implant-per-patient technique" can be applied. CONCLUSION: The number of intraoral radiographs per examination and, more importantly, radiographic examinations can be reduced without jeopardizing good clinical management, a statement valid even for Branemark implants with advanced bone loss. PMID- 18384399 TI - Immediate and early loading of oxidized tapered implants in the partially edentulous maxilla: a 1-year prospective clinical, radiographic, and resonance frequency analysis study. AB - BACKGROUND: The use of immediate/early implant loading protocols offers obvious advantages for the patient. Although well documented in the totally edentulous mandible, information about clinical outcomes from such protocols in the partially edentate maxilla is lacking. PURPOSE: The present study was conducted to clinically and radiographically evaluate a tapered implant design with an oxidized surface for immediate/early loading in the partially edentulous maxilla. The aim was also to correlate implant stability measurements using resonance frequency analysis (RFA) with implant diameter and length, bone quality and quantity, and marginal bone levels and marginal bone loss. MATERIALS AND METHODS: A total of 32 patients with a need of implant treatment in their partially edentulous maxilla were included in the study. A total of 53 Replace Select TiUnite implants (Nobel Biocare AB, Goteborg, Sweden) were used in the study; 16 for single tooth replacements in 16 patients and 37 implants for partial bridges in another 16 patients. The single tooth replacements were loaded the same day with a temporary crown, while permanent partial bridges were delivered within 16 days. Intraoral radiographs were taken at surgery and after 1 year for marginal bone measurements. RFA measurements were performed at baseline and after 3, 6, and 12 months. RESULTS: One implant used for a single tooth replacement failed, giving an overall survival rate of 98.1% after 1 year. On average, 1.1 mm (SD 1.0) bone was lost during 1 year; 1.5 mm (SD 1.0) in single tooth and 0.9 mm (SD 1.0) in partial cases. The implant stability increased with time from 63.3 implant stability quotient (ISQ) (SD 6.1) at baseline to 64.3 (SD 5.3), 65.0 (SD 4.6), and 66.8 (SD 5.6) after 3, 6, and 12 months, respectively. The average change from baseline to 1 year was 3.3 ISQ (SD 5.0) and was statistically significant (p < .05). There was no difference between single and partial cases. Implant stability correlated with bone quantity and quality at implant sites, but not with marginal bone level measurements. CONCLUSIONS: It is concluded that immediate/early loading can be used in the partially edentulous maxilla with good clinical and radiographic short-term outcomes. Implant stability at placement correlated with bone quantity and quality, and increased with time as measured with RFA, indicating a favorable bone tissue response to the loaded implants. Any correlations between RFA and marginal bone level measurements were not observed in the present study. PMID- 18384400 TI - Influence of defect depth on resonance frequency analysis and insertion torque values for implants placed in fresh extraction sockets: a human cadaver study. AB - BACKGROUND: Clinical studies show promising outcomes with implants inserted at the time of extraction. However, this often results in an initial bone defect at the marginal region which preferably should heal for an optimal function. Therefore, monitoring of these implants is vital. PURPOSES: The aims of this study were to determine the initial stability of implants placed into fresh extraction sockets, and to explore the correlations between the peri-implant bone levels and implant stability parameters. MATERIALS AND METHODS: Six human cadaver mandibles including all natural teeth were selected for this study. All natural teeth were gently extracted, and 84 implants were immediately placed into fresh extraction sockets with five different implant depths. The maximum insertion torque values were recorded, and primary implant stability measurements were performed by means of resonance frequency analysis (RFA). The vertical distance between implant/abutment junction and the first bone-implant contact was recorded using a periodontal probe. RESULTS: It was found that the insertion torque and RFA were 28.9 +/- 7 Ncm and 65.6 +/- 9 implant stability quotient (ISQ), respectively, for 420 measurements from all 84 implants. Statistically significant correlation was found between insertion torque and ISQ values (r = 0.86; p < .001) for all implants. Both insertion torque and ISQ values dramatically decreased when the amount of peri-implant vertical bone defect increased. CONCLUSION: The results of this study demonstrated a linear relationship between peri-implant vertical bone defect depth and RFA value. It is proposed that the RFA method is sensitive to detect changes of the marginal bone level and may be used to monitor healing of peri-implant bone defects. PMID- 18384401 TI - Immediate loading of implants in the edentulous maxilla: use of an interim fixed prosthesis followed by a permanent fixed prosthesis: a 32-month prospective radiological and clinical study. AB - PURPOSE: The aim of this study was to prospectively evaluate the survival rate of splinted and immediately loaded Straumann sandblasted, large-grit, acid-etched, solid-screw dental implants in the edentulous maxilla after 32 months of loading. MATERIALS AND METHODS: Twenty-eight patients (mean age 63 years) with edentulous maxillae received 168 implants (six each) and an implant-supported fixed interim prosthesis within 24 hours after surgery. After a mean healing time of 15 weeks, the patients received permanent screw-retained prostheses. Clinical and radiological examinations were made at implant placement and after 8, 20, and 32 months of loading. All permanent prostheses were removed at the 32-month follow up; implant stability was checked with a torque device, and the implant stability quotient was determined with resonance frequency analysis. RESULTS: Mean marginal bone loss from baseline to 8 months after loading was 1.6 mm (SD 1.16; p = .094), from 8 to 20 months 0.41 mm (SD 0.63; p = .094), and from 20 to 32 months 0.08 mm (SD 0.49; p = .039). The 32-month cumulative survival rate was 98.2%. CONCLUSIONS: The 32-month survival of solid-screw implants - immediately loaded within 24 hours after placement - was similar to survival rates reported for solid-screw implants with conventional loading. Immediate loading and splinting of implants in the edentulous maxilla is a viable treatment alternative. PMID- 18384402 TI - Chronological changes in the ultrastructure of titanium-bone interfaces: analysis by light microscopy, transmission electron microscopy, and micro-computed tomography. AB - PURPOSES: The objectives of this study were to chronologically examine the titanium-bone interfaces and to clarify the process of osseointegration using light microscopy, transmission electron microscopy (TEM), and micro-computed tomography (CT). MATERIALS AND METHODS: Experimental implants (Ti-coating plastic implants) were placed into tibiae of 8-week-old rats. Animals were sacrificed at 1 to 28 days after implant placement and prepared tissue specimens for a light microscope, a TEM, and micro-CT. RESULTS: New bone formation began 5 days after implant placement, and osseointegration was obtained by 14 days after implant placement. Osseointegration was well developed by 28 days after implant placement. DISCUSSION: TEM and quantitative computer tomography (QCT) results indicated that bone formation in osseointegration of titanium implants did not occur from the surfaces of the implant or preexisting bone, but it was likely that bone formation progressed at a site a small distance away from the surface. The bone formation took place in a scattered manner. Small bone fragments adhered to each other and transformed into reticular-shaped bone, and finally these bones became lamellar bone. CONCLUSION: Comparative analysis of the titanium-bone interfaces using light microscopy, TEM, and QCT by micro-CT revealed the precise process of osseointegration. PMID- 18384404 TI - Anatomical bases for the insertion of zygomatic implants. AB - PURPOSE: The purpose of this study was to obtain anatomical information for installing four zygomatic implants, by measurements of maxillae and zygomata. MATERIALS AND METHODS: Linear and angular measurements were obtained from the maxillae and zygomata of 40 dry skulls. RESULTS: Based on mean and standard deviation values, the installation angle of the additional implant in the four zygoma technique was between 25 degrees and 47 degrees , and that of the conventional zygomatic implant was between 39 degrees and 62 degrees . The distance between the alveolar crest in the canine region and the lateral margin of the orbital socket was 53.42 mm, and between the alveolar crest in the premolar region and the area closest to the lateral margin of the orbital socket was 42.47 mm. CONCLUSIONS: When the angle of installation of the additional implant is less than 25 degrees and that of the conventional zygomatic implant is less than 39 degrees , perforation of the maxilla, zygoma, or the infratemporal fossa must be avoided. When the angle of installation of an additional implant is greater than 47 degrees and that of the conventional zygomatic implant is greater than 62 degrees , perforation of the orbital floor must be avoided. The length of an additional implant was greater than the length of a conventional zygomatic implant, and should be measured prior to drilling, because the greatest value found for this distance was 61.94 mm, while the maximum length of the zygomatic implants currently available on the market is 52.5 mm. PMID- 18384403 TI - Marginal bone loss at implants: a retrospective, long-term follow-up of turned Branemark System implants. AB - BACKGROUND: Lately, presence of progressive bone loss around oral implants has been discussed. PURPOSE: The aim of this study was to report in a large patient group with different prosthetic restorations marginal bone level and its change as measured in radiographs obtained from prosthesis insertion up to a maximum 20 years in service. Further, it also aimed to study the impact of gender, age, jaw, prosthetic restoration, and calendar year of surgery. MATERIALS AND METHODS: Out of 1,716 patients recorded for clinical examination during 1999, 1,346 patients (78.4%) could be identified. A total of 640 patients (3,462 originally installed Branemark System implants, Nobel Biocare, Goteborg, Sweden) with a follow-up of >or=5 years were included in the study, while patients with continuous overdentures and augmentation procedures were not. Distance between the fixture/abutment junction (FAJ) and the marginal bone level was recorded. RESULTS: The number of implants with a mean bone level of >or=3 mm below FAJ increased from 2.8% at prosthesis insertion to 5.6% at year 1, and 10.8% after 5 years. Corresponding values after 10, 15, and 20 years were 15.2, 17.2, and 23.5%, respectively. Implant-based bone loss was as a mean 0.8 mm (SD 0.8) after 5 years, followed by only minor average changes. Mean bone loss on patient level followed a similar pattern. Disregarding follow-up time, altogether 183 implants (107 patients) showed a bone loss >or=3 mm from prosthesis insertion to last examination. Significantly larger bone loss was found the older the patient was at surgery and for lower jaw implants. CONCLUSIONS: Marginal bone support at Branemark implants was with few exceptions stable over years. PMID- 18384405 TI - Immediate rehabilitation of the mandible with fixed full prosthesis supported by axial and tilted implants: interim results of a single cohort prospective study. AB - PURPOSE: The aims of this prospective study were to assess the treatment outcome of immediately loaded full-arch fixed bridges anchored to both tilted and axially placed implants for the rehabilitation of the mandible and to compare the outcome of axial versus tilted implants. MATERIALS AND METHODS: Sixty-two patients (34 women and 28 men) were included in the study. Each patient received a full-arch fixed bridge supported by two axial implants and two distal tilted implants (All on-Four, Nobel Biocare AB, Goteborg, Sweden). Loading was applied within 48 hours of surgery. Patients were scheduled for follow-up at 6, 12, 18, and 24 months, and annually up to 5 years. At each follow-up, plaque level and bleeding scores were assessed; moreover, patient's satisfaction for aesthetics and function was evaluated by a questionnaire. Radiographic evaluation of marginal bone level change was performed at 1 year. RESULTS: The overall follow-up range was 6 to 43 months (mean 22.4 months). Forty-four patients were followed for a minimum of 1 year. No implant failures were recorded to date, leading to a cumulative implant survival and prosthesis success rate of 100%. Plaque level and bleeding scores showed progressive decrease over time, parallel to increase of satisfaction for both aesthetics and function. No significant difference in marginal bone loss was found between tilted and axial implants at 1-year evaluation. CONCLUSION: The present preliminary data suggest that immediate loading associated with tilted implants could be considered a viable treatment modality for the mandible. PMID- 18384406 TI - Effect of hydroxyapatite and titania nanostructures on early in vivo bone response. AB - PURPOSE: Hydroxyapatite (HA) or titania nanostructures were applied on smooth titanium implant cylinders. The aim was to investigate whether nano-HA may result in enhanced osseointegration compared to nano-titania structures. MATERIALS AND METHODS: Surface topography evaluation included detailed characterization of nano size structures present at the implant surface combined with surface roughness parameters at the micro- and nanometer level of resolution. Microstructures were removed from the surface to ensure that bone response observed was dependent only on the nanotopography and/or chemistry of the surface. Early in vivo histological analyses of the bone response (4 weeks) were investigated in a rabbit model. RESULTS: In the present study, nano-titania-coated implants showed an increased coverage area and feature density, forming a homogenous layer compared to nano-HA implants. Bone contact values of the nano-titania implants showed a tendency to have a higher percentage as compared to the nano-HA implants (p = .1). CONCLUSION: Thus, no evidence of enhanced bone formation to nano-HA-modified implants was observed compared to nano-titania-modified implants. The presence of specific nanostructures dependent on the surface modification exhibiting different size and distribution did modulate in vivo bone response. PMID- 18384407 TI - Clinical and microbiological determinants of ailing dental implants. AB - BACKGROUND: The failure of the host tissue to establish or maintain osseointegration around dental implants is due to either occlusal or parafunctional forces, premature loading, ill-directed stress, or microbial infection. The long-term failure rate of dental implants is generally 5-10%. Although a variety of etiologies of early peri-implant bone loss (from implant placement to 1-year post-loading) have been proposed, factors associated with late implant failures are less well understood but are probably related to both the peri-implant microbial environment and host factors. Discriminating between causes of implant failure is of importance for instituting a successful implant therapy. PURPOSE: The objective of this cross-sectional split-mouth study was to identify clinical, radiographic, and bacterial characteristics of peri-implant disease sites. MATERIALS AND METHODS: Fifteen patients with bilateral implants (Branemark, Nobel Biocare AB, Goteborg, Sweden; and 3i implant systems, Implant Innovations Inc., Palm Beach Gardens, FL, USA) participated in the study. Sites with peri-implant (radiographic bone loss beyond the third implant thread) and peri-implant healthy tissues (radiographic bone level above the first implant thread) were identified in periapical radiographs using a long-cone paralleling projection technique. Microbiological identification was carried out using established anaerobic culture techniques. A descriptive statistics based on means and standard deviations was reported. RESULTS: Peri-implant bone loss was associated with the absence of radiographic crestal lamina dura, peri-implant pocket depth, pain on chewing, and the submucosal presence of the putative periodontopathogens Tannerella forsythia, Campylobacter species, and Peptostreptococcus micros. Pain was associated with P. micros, Fusobacterium species, and Eubacterium species. DISCUSSION AND CONCLUSION: The absence of radiographic crestal lamina dura and the presence of suspected major periodontal pathogens seem to be associated to peri-implantitis. PMID- 18384408 TI - Determination of bone quality of 372 implant recipient sites using Hounsfield unit from computerized tomography: a clinical study. AB - BACKGROUND: The type and architecture of bone are very important factors in the successful implant treatment, and it is manifested that higher implant failure is more likely in the poorer quality of bone. Conventional bone classifications have recently been questioned because they are subjective and retrospective. PURPOSE: This clinical study aimed to determine the variations of the bone density in dental implant recipient sites using computerized tomography (CT). MATERIALS AND METHODS: The study group comprised of randomly selected 140 patients with 372 implant sites. Recipient sites for implant placement were determined based on CT data using implant planning StentCad software (Media Lab Software, La Spezia, Italy). The mean bone density values in Hounsfield unit (HU) of the simulated implant areas were recorded using the StentCad software. RESULTS: The HU values ranged from 68 to 1,603 HU. It was found that mean bone density values were 927 +/- 237, 721 +/- 291, 708 +/- 277, and 505 +/- 274 HU in the anterior mandible, posterior mandible, anterior maxilla, and posterior maxilla, respectively. CONCLUSION: Preoperative CT examination may be a useful method for determining the bone density of recipient areas before implant placement, and this valuable information about bone quality helps clinicians to make better treatment planning regarding the implant positions. PMID- 18384409 TI - Two alternative surgical techniques for enhancing primary implant stability in the posterior maxilla: a clinical study including bone density, insertion torque, and resonance frequency analysis data. AB - BACKGROUND: The primary stability of dental implants associated with resistance to micromotion during healing is affected by surgical technique and implant design, which are important especially in the soft bone, where implant failures are more likely. PURPOSES: This study was designed to compare the parameters associated with implant insertion using two different methods of enhancing implant primary stability and to identify any relationship between these parameters at implant insertion. MATERIALS AND METHODS: A total of 60 implants were placed in the maxillary posterior regions of 22 patients. The bone densities at the implant sites were recorded using a computerized tomography machine in Hounsfield unit (HU). The maximum insertion torque data were recorded with the Osseocare (Nobel Biocare AB, Goteborg, Sweden) equipment, while resonance frequency analysis (RFA) measurements were taken using an Osstell (Integration Diagnostics AB, Goteborg, Sweden) machine at implant surgery. Comparisons including HU, Ncm, and implant stability quotient were made between two control groups (C1 and C2), and corresponding four test groups (T1-T4) using thinner drills to enhance primary implant stability. RESULTS: Two implants were lost, meaning an overall implant survival rate of 96.6% after 3 +/- 1 years. When compared to control groups, significantly higher mean maximum insertion torque and RFA values were found for corresponding test groups. In addition, strong correlations were observed between the bone density and insertion torque, and implant stability values at implant placement. CONCLUSION: The results of this study suggest that using thinner drills for implant placement in the maxillary posterior region where bone quality is poor may improve the primary implant stability, which helps clinicians to obtain higher implant survival rates. PMID- 18384410 TI - Accuracy of linear measurement provided by cone beam computed tomography to assess bone quantity in the posterior maxilla: a human cadaver study. AB - PURPOSE: The aim of this study was to assess, for implant placement in the posterior maxilla, the accuracy of linear measurements provided by cone beam computed tomography (CBCT) using an image intensifier tube and television (TV) chain as an X-ray detector despite a loss of contrast resolution. The NewTom 9000 (Quantitative Radiology, Verona, Italy) was used to explore the posterior maxilla. MATERIALS AND METHODS: Fourteen measurements were taken in three dry maxillaries. On every anatomical site, three fiducial markers were placed on the bony crest to define a plane. Dry maxillaries were submitted to CBCT imaging examination. The maxillaries were then sawn according to the previously defined planes, and bone height and width were assessed using a caliper. The same measurements were taken on images. RESULTS: Clinical analysis demonstrated no difference between real measurements and image measurements. CONCLUSIONS: Although cadaver bone density may not correspond to the density of vital bone, this in vitro study indicates that CBCT images provided by technique using image intensifier tube and TV chain as an X-ray detector are reliable to define the bone volume of the posterior maxilla for the purpose of planning the implant axis. PMID- 18384411 TI - Implant treatment of patients with edentulous jaws: a 20-year follow-up. AB - BACKGROUND: Implant-supported prostheses are today often used in rehabilitation of partially or totally edentulous patients. Both patients and the dental profession often regard implant treatment as successful in a life perspective. Therefore, studies with a long-term follow-up are important. PURPOSE: The aim was to investigate the outcome of implant treatment with fixed prostheses in edentulous jaws after 20 years, with special reference to survival rate of implants and prostheses and frequency of peri-implantitis. MATERIALS AND METHODS: The patient material was a group of patients treated in the early 1980s. The original patient group comprised the first 48 consecutive patients treated with implant-supported prostheses at Umea University. All patients were edentulous in one or two jaws. The patients had a mean age at the implant insertion of 54.3 years (range 40-74). At the planning of this study 20 years after treatment, 19 of the 48 patients were found to be deceased. Of the 29 patients still alive, 21 patients with altogether 23 implant-supported prostheses could be examined clinically and radiographically. All patients were treated ad modum Branemark (Nobel Biocare AB, Goteborg, Sweden) with a two-stage surgical procedure. The implants had a turned surface. Abutment connections were performed 3 to 4 months after fixture insertion in the mandible, and after a minimum of 6 months in the maxilla. The prostheses were fabricated with a framework of gold alloy and acrylic artificial teeth. RESULTS: The 21 patients (with 23 implant prostheses) examined had at the time of treatment got 123 implants (27 in the upper jaw and 96 in the lower jaw) inserted. Only one of these implants had been lost (about 2 years after loading) giving a survival rate of 99.2%. Very small changes occurred in the marginal bone level. Between the 1 and 20-year examinations, the mean bone loss was 0.53 mm and the mean bone level at the final examination was 2.33 mm below the reference point. CONCLUSIONS: This follow-up over two decades of implant-supported prostheses demonstrates a very good prognosis for the treatment performed. The frequencies of peri-implantitis, implant failures, or other complications were very small, and the original treatment concept with a two stage surgery and a turned surface of the implants will obviously give very good results. PMID- 18384412 TI - Clinical outcome of mini-screws used as orthodontic anchorage. AB - BACKGROUND: Implants and orthodontics are an important combination to obtain intraoral anchorage and eliminate the disadvantages linked with extraoral anchorage such as compliance problems, aesthetical, and social factors. The mini screw is a simple, relatively low-cost method to provide intraoral anchorage. PURPOSES: The aims of this study were to evaluate clinical success and longevity of mini-screws during orthodontic treatment and to assess the patient's opinion. MATERIALS AND METHODS: Fifty mini-screws were inserted in the mandible and maxilla of 21 patients with a flapless technique under local anesthesia. The patients were recalled after 2 weeks and from then on every other 2 months, and periodontal parameters and stability of the screws were evaluated at regular intervals. Patients received a questionnaire to assess their opinion regarding the treatment. RESULTS: Thirty-three mini-screws (64%) remained stable sufficiently long to obtain the effect during the orthodontic movement. The survival was comparable in mandible or maxilla, and not related to the orthodontic forces applied or time of activation of the load. The results do suggest that a waiting period of 1 week before loading improves success, and mini screws inserted into the anterior region score better also compared to the posterior region. Initial periodontal parameters, which are very important in prognosis of orthodontic treatment, are not influencing the success rate in the examined group. Patients complained in 40-50% of the cases of pain during or after surgery, but this did not negatively affect the final general satisfaction with the treatment. CONCLUSION: The mini-screw implant is an easy and an inexpensive method for temporary anchorage of orthodontic appliances. The functioning time is short, however, and retreatment may often be required. PMID- 18384413 TI - ICHD-II diagnostic criteria for Tolosa-Hunt syndrome in idiopathic inflammatory syndromes of the orbit and/or the cavernous sinus. AB - A bibliographical search was conducted for papers published between 1999 and 2007 to verify the validity of International Classification of Headache Disorders (ICHD)-II criteria for the Tolosa-Hunt syndrome (THS) in terms of (i) the role of magnetic resonance imaging (MRI); (ii) which steroid treatment should be considered as adequate; and (iii) the response to treatment. Of 536 articles, 48, reporting on 62 patients, met the inclusion criteria. MRI was positive in 92.1% of the cases and it normalized after clinical resolution. There was no evidence of which steroid schedule should be considered as adequate; high-dose steroids are likely to be more effective both to induce resolution and to avoid recurrences. Pain subsided within the time limit required by the ICHD-II criteria, but signs did not. We conclude that THS diagnostic criteria can be improved on the basis of currently available data. MRI should play a pivotal role both to diagnose and to follow-up THS. PMID- 18384414 TI - Spontaneous vertebral artery dissection mimicking cluster headache. PMID- 18384415 TI - Idiopathic intracranial hypertension with and without papilloedema in a consecutive series of patients with chronic migraine. AB - Chronic migraine (CM) has been associated with idiopathic intracranial hypertension without papilloedema (IIHWOP), a significant percentage of these cases occurring in obese patients with intractable headache. A prospective study from February 2005 to June 2006 was made of 62 CM patients who fulfilled International Headache Society diagnostic criteria and had cerebral magnetic resonance venography (MRV) and lumbar puncture (LP) done. Two patients were excluded, six (10%) with elevated cerebrospinal fluid (CSF) open pressure (OP), five with body mass index (BMI) > 25. None of the patients had papilloedema or abnormal MRV. BMI and CSF OP were significantly correlated (r = 0.476, P < 0.001, Pearson's correlation test). Obesity (defined as BMI > 30) was a predictor of increase in intracranial pressure (defined as OP > 200 mmH(2)O) (f = 17.26, 95% confidence interval 6.0, 8.6; P < 0.001). From our study we strongly recommend that not only intractable CM patients with high BMI, but also first diagnosed patients with BMI > 30 should be systematically evaluated by a LP to rule out IIHWOP. PMID- 18384416 TI - Ophthalmoplegic migraine with paresis of the sixth nerve: a neurovascular compression syndrome? PMID- 18384417 TI - Idiopathic trigeminal neuropathy may respond to greater occipital nerve injection. PMID- 18384418 TI - Familial hemiplegic migraine type 1 shows no hypersensitivity to nitric oxide. AB - Familial hemiplegic migraine type 1 (FHM-1) is a dominantly inherited subtype of migraine with aura and transient hemiplegia associated with mutations in the CACNA1A gene. FHM-1 shares many phenotypical similarities with common types of migraine, indicating common neurobiological pathways. Experimental studies have established that activation of the nitric oxide-cyclic guanosine monophosphate (NO-cGMP) pathway plays a crucial role in migraine pathophysiology. Therefore, we tested the hypothesis that CACNA1A mutations in patients with FHM-1 are associated with hypersensitivity to NO-cGMP pathway. We included eight FHM-1 patients with R583Q and C1369Y mutations and nine healthy controls, who received intravenous infusions of 0.5 microg kg(-1) min(-1) glyceryl trinitrate (GTN) over 20 min. We recorded: headache intensity on a verbal rating scale; mean flow velocity in the middle cerebral artery (V(meanMCA)) by transcranial Doppler; diameter of the superficial temporal artery (STA) by Dermascan. One patient reported migraine without aura 5 h after start of the GTN infusion. No aura was reported. The AUC(headache) in the immediate phase was more pronounced in patients than in controls (P = 0.01). In the 14 h following GTN infusion, there was no difference in the AUC(headache) between patients and controls (P = 0.17). We found no difference in the AUC(VmeanMCA) (P = 0.12) or AUC(STA) (P = 0.71) between FHM-1 patients and controls. None of the control persons reported migraine-like headache. FHM-1 patients do not show hypersensitivity of the NO cGMP pathway, as characteristically seen in migraine patients with and without aura. This indicates that the pathophysiological pathways underlying migraine headache in FHM-1 may be different from the common types of migraine. PMID- 18384419 TI - Comparison of CGRP and NO responses in the human peripheral microcirculation of migraine and control subjects. AB - Calcitonin gene-related peptide (CGRP) and nitric oxide (NO) are two molecules shown to have a role in migraine pathophysiology. Our objective was to test the hypothesis that migraine subjects are particularly sensitive to these signal molecules. The cutaneous microvascular responses to endothelial and non endothelial dependent dilators were tested using laser Doppler flowmetry in combination with iontophoresis. The blood flow responses to iontophoretic administration of the endothelium-dependent vasodilator acetylcholine (ACh), or to the endothelium-independent dilators sodium nitroprusside (SNP) and CGRP, and to local warming (44 degrees C) were compared in this controlled trial. The design was that of two arms: patients diagnosed with migraine without aura (n = 9) for >10 years were compared with nine healthy subjects matched for age and gender (seven female and two male, age range 30-60 years). Iontophoretic administration resulted in local vasodilation. ACh induced a relaxation of 1225 +/- 245% (relative to baseline) in controls and 1468 +/- 368% (P > 0.05) in migraine. The responses to SNP were 873 +/- 193% in controls and 1080 +/- 102% (P > 0.05) in migraine subjects. The responses to CGRP were 565 +/- 89% in controls and 746 +/- 675% (P > 0.05) in migraine patients. The responses to local heating which induced maximum dilation did not differ between the groups (1976 +/- 314% for controls and 1432 +/- 226% in migraine; P > 0.05. We conclude that there is no change in the microvascular responsiveness of the subcutaneous microvasculature in migraine. PMID- 18384420 TI - Validation of the revised Patient Perception of Migraine Questionnaire (PPMQ-R): measuring satisfaction with acute migraine treatment in clinical trials. AB - This study was aimed to evaluate in clinical trial settings the psychometric properties of the revised Patient Perception of Migraine Questionnaire (PPMQ-R), a satisfaction measure for acute migraine treatment. The PPMQ-R was administered 24 h post dosing in 1304 migraineurs randomized to two identical Phase 3, single attack trials. Reliability, concurrent and construct validity and known-groups validity were evaluated using Cronbach's alpha, Pearson correlations and analysis of variance, respectively. PPMQ-R scale and Total scores (Efficacy, Functionality and Ease of use) showed very good internal consistency reliability (alpha 0.84 0.99). Efficacy, Functionality and Total PPMQ-R scores showed large, inverse relationships with migraine pain severity, number of migraine symptoms and work ability (r = -0.62 to -0.75; all P < 0.0001). All scales discriminated among migraine pain severity levels (all P < 0.001). The PPMQ-R has sufficient evidence of validity and reliability for measuring patient satisfaction, an important benchmark of quality and effective care. PMID- 18384421 TI - Sacrococcygeal pilonidal disease. AB - BACKGROUND: Sacrococcygeal pilonidal is a common disease in active young adults. Many surgical methods have been proposed, although no clear consensus as to the optimal treatment has been reported. This review looks at the different surgical techniques available and examines the reported results of primary healing, recurrent disease and complications (including delayed healing). METHOD: A literature search using the Medline database was performed to locate English language articles on surgery for pilonidal disease. Further articles were obtained from the references cited in the literature initially reviewed. RESULTS: Management should be tailored according to the individual and whether the disease is acute or chronic. Treatment should take into consideration hospital stay and return to work. Simple excision, curettage, partial lateral wall excision, or marsupialisation, are simple techniques with good results. They can be used for the initial surgery but their use is not recommended for recurrent disease. The modified rhomboid flap for recurrent disease has consistently shown positive results in terms of complication rates and recurrence. CONCLUSION: We would recommend tailored treatment with simple excision for initial presentation and the modified rhomboid flap for recurrent disease. PMID- 18384422 TI - Awareness of risk factors amongst first degree relative patients with colorectal cancer. AB - BACKGROUND: The association between family history of colorectal cancer (CRC) in first degree relatives (FDRs) and the risk of developing the disease has been well established. However, little is known about the degree of awareness of CRC risk factors amongst FDRs. This study examined awareness of familial, dietary and lifestyle risk amongst FDRs. METHOD: Consent to approach relatives was obtained from CRC patients who attended outpatients following surgery for CRC. A study specific questionnaire was sent by post to their FDRs. The dietary and lifestyle risk factors tested in the questionnaire were those promoted in CRC educational material. RESULTS: One hundred and four questionnaires were returned out of 137 sent (76%). Fifty-nine per cent (n = 61) of FDRs were aware of their increased risk. Fifty-five per cent (n = 57) were aware of the link between CRC and smoking; however, knowledge of association of CRC with excessive alcohol consumption (32%) and lack of exercise (40%) was less well appreciated. Higher awareness scores were obtained by those with a university education (P < 0.001). Ninety four per cent believed that relatives of CRC patients were not provided with enough information as to their being at risk and risk factors. CONCLUSIONS: This study provides an initial assessment of the awareness of CRC risk factors in a population at increased risk. It demonstrates a poor level of knowledge. Improved education strategies allowing FDRs to make dietary and lifestyle modifications that could reduce their lifetime risk of developing CRC are necessary. PMID- 18384423 TI - Laparoscopic colonic surgery in Denmark 2004-2007. AB - OBJECTIVE: Laparoscopic colonic surgery was introduced about 15 years ago and has together with the evidence-based 'fast-track' methodology improved early postoperative outcome. The purpose of this study was to asses the organization and early outcome after laparoscopic colonic surgery in Denmark from 2004 to 2007. METHOD: Based upon the National Patient Register, all laparoscopic colonic operations performed in Denmark between January 2004 and December 2006 were analysed regarding number of hospital departments and procedures, hospital stay, readmissions and mortality. RESULTS: One thousand one hundred and forty-nine laparoscopic colonic resections without simultaneous stoma formation were performed in the study period. Twenty-five departments performed the procedures but only four departments performed more than 100 procedures. The median length of primary stay was 4 days (mean 7.7 days). One hundred and twenty-five (10.9%) patients were re-admitted within 30 days and total length of stay (primary plus readmissions) was a median of 5 days (mean 8.5 days). Thirty-day mortality was 2.6% and hospital mortality 3.5%. CONCLUSION: This nationwide study has shown an increased implementation of laparoscopic colonic surgery but probably performed in too many low volume departments. Laparoscopic colonic surgery should be monitored and further advances secured by adjustment of perioperative care to fast-track care. PMID- 18384424 TI - Imbrication of the external anal sphincter may yield similar functional results as overlapping repair in selected patients. AB - OBJECTIVE: Overlapping external anal sphincter repair is the preferred procedure for incontinent patients with functional yet anatomically disrupted anterior external anal sphincter. When incomplete disruption, thinning or technically difficult mobilization of the external anal sphincter occurs, imbrication without division may be the more feasible surgical option. The aim of the study was to assess retrospectively the indications for external anal sphincter imbrication in patients who underwent either overlapping external anal sphincter repair or external anal sphincter imbrication, and to compare the success rates. METHOD: Patients who had external anal sphincter repair and follow up of at least 3 months were studied. Fecal incontinence was measured using the validated Wexner fecal Incontinence Scoring system (0 = perfect continence, 20 = complete incontinence); postoperative scores 0-10 were interpreted as successful, and scores of 11-20 as failures. RESULTS: A total of 131 females who had anal sphincter repair between 1988 and 2000 were analysed. One hundred and twenty-one patients had overlapping external anal sphincter repair (group I), and 10 had external anal sphincter imbrication (group II). Indications for external anal sphincter imbrication were attenuation of the external anal sphincter without overt defect (n = 5), partial disruption of external anal sphincter with muscle fibres bridging the scar (n = 2), thick bulk of scar between the muscle edges (n = 2), and wide lateral retraction of the muscle edges (n = 1). There were no statistically significant differences between the groups relative to preoperative incontinence score (16.5 vs 16.5, P = 0.99), pudendal nerve terminal motor latency assessment (left 9.6%vs 0.0%, P = 0.19; right 13.4%vs 11.1%, P = 0.84), and extent of electromyography pathology (61%vs 47%, P = 0.30). The patients in group I were younger than those in group II (mean age 50.8 years vs. 61.7 years, respectively; P = 0.052) and the length of follow-up was significantly longer (32.3 months vs 14.3 months, respectively; P < 0.0001). Both procedures had similar success rates (59.5%vs 60%; P = 0.98). CONCLUSION: Imbrication of the external anal sphincter may yield similar results as overlapping external anal sphincter repair in patients with incomplete external anal sphincter disruptions, external anal sphincter attenuation, and in patients presenting with wide lateral retraction of the muscle edges. PMID- 18384425 TI - National UK audit of procedure for prolapsing haemorrhoids on behalf of the Association of Coloproctology of Great Britain and Ireland. AB - OBJECTIVE: To assess the safety and short term outcomes of the procedure for prolapsing haemorrhoids (PPH), a relatively new procedure for the treatment of symptomatic haemorrhoids. METHOD: In 2005, the Association of Coloproctology of Great Britain and Ireland set up an online electronic database to audit the indications and outcomes for patients undergoing a PPH procedure. RESULTS: During the audit period, 695 patients were entered onto the database by 61 surgeons (range 1-50 patients per surgeon). The main indications for surgery were bleeding (90.5%) and prolapse (83.9%). Three hundred and ninety-seven (57.1%) patients had grade III or IV haemorrhoids. PPH was performed under general anaesthetic in 602 (86.6%) cases and a consultant surgeon performed the procedure in 572 (82.3%) cases. The median length of stay was 1 day (range 0-6 days). Two hundred and eighty-nine (41.6%) procedures were performed as a day case. Immediate complications were recorded in 75 (10.8%) patients, the commonest being bleeding (21) and urinary retention (24). At 6-week follow-up, 626 (90.1%) patients were pain free. Five patients required hospital re-admission for secondary haemorrhage (3), peri-anal abscess (1) and pain (1). The commonest problems were minor bleeding (48), urgency (22), pain (14), continued prolapse (12) and pruritus (11). Four patients required an open haemorrhoidectomy for persistent symptomatic haemorrhoids. CONCLUSION: Procedure for PPH is a safe and effective procedure for symptomatic haemorrhoids with good short-term outcomes. Long-term follow-up is required perhaps through a compulsory national register. PMID- 18384426 TI - ATM germline mutations in Spanish early-onset breast cancer patients negative for BRCA1/BRCA2 mutations. AB - Heterozygous carriers of ATM (ataxia telangiectasia mutated gene) mutations have increased risk of breast cancer (BC). We have estimated the prevalence of mutations in the ATM gene among Spanish patients with early-onset BC. Forty-three patients diagnosed with BC before the age of 46 years, and negative for BRCA1 and BRCA2 mutations, were analysed for the presence of ATM mutations. A total of 34 ATM sequence variants were detected: 1 deleterious mutation, 10 unclassified variants and 23 polymorphisms. One patient (2.3%) carried the ATM deleterious mutation (3802delG that causes ataxia telangiectasia in the homozygous state) and 13 patients carried the 10 ATM unclassified variants. The truncating mutation 3802delG and eight of the rare variants were not detected in a control group of 150 individuals. Different bioinformatic sequence analysis tools were used to evaluate the effects of the unclassified ATM changes on RNA splicing and function protein. This in silico analysis predicted that the missense variants 7653 T>C and 8156 G>A could alter the splicing by disrupting an exonic splicing enhancer motif and the 3763 T>G, 6314 G>C, and 8156 G>A variants would affect the ATM protein function. These are the initial results concerning the prevalence of germline mutations in the ATM gene among BC cases in a Spanish population, and they suggest that ATM mutations can confer increased susceptibility to early onset BC. PMID- 18384427 TI - Gene dosage imbalances in patients with 46,XY gonadal DSD detected by an in-house designed synthetic probe set for multiplex ligation-dependent probe amplification analysis. AB - The development of a testis requires the proper spatiotemporal expression of the SRY gene and other genes that act in a dosage-sensitive manner. Mutations in the SRY gene account for only 10-15% of patients with 46,XY gonadal disorder of sex development (DSD). To enable the diagnostics of deletions and duplications of genes known to be involved in different forms of DSD, we developed a synthetic probe set for multiplex ligation-dependent probe amplification (MLPA) analysis. Here, we report the results from the analysis of 22 patients with 46,XY gonadal DSD. The analysis with the DSD probe set has led to the identification of two copy number variations, an 800-kb NR0B1 (DAX1) locus duplication on Xp21 in a patient with isolated partial gonadal dysgenesis and a duplication of the SRD5A2 gene that represents a rare normal variant. The described MLPA kit represents an optimal complement to DNA sequence analysis in patients with DSD, enabling screening for deletions and duplications of several genes simultaneously. Furthermore, the second identification of an NR0B1 locus duplication in a patient with isolated gonadal dysgenesis, without dysmorphic features and/or mental retardation, highlights the importance of evaluating NR0B1 duplication in patients with gonadal dysgenesis. PMID- 18384428 TI - Detection of chromium allergy by cellular in vitro methods. AB - BACKGROUND: The standard assay for the detection of chromium sensitization, the patch test, does not allow discrimination between patients with and without clinical symptoms of allergy. OBJECTIVE: The aim of this study was to prove whether cellular in vitro tests are predictive of chromium allergy. METHODS: Chromium-sensitized volunteers with and without clinically manifest allergy and non-sensitized healthy controls (n=37, 19, and 26, respectively) were analysed by cellular in vitro methods using tri- and hexavalent chromium (chromium chloride and potassium dichromate) as stimuli. The results were correlated with clinical and anamnestic data. RESULTS: Sensitized individuals with an allergy displayed significantly higher lymphocyte transformation test (LTT) responses than sensitized volunteers without allergy and controls (P<0.05 and P<0.01, respectively). 12.5 microg/mL of chromium chloride and 50 ng/mL of potassium dichromate were found to be optimal to discriminate between sensitized individuals with and without allergy. Combining the results of chromium chloride and potassium dichromate LTT, a positive reaction to at least one of the stimuli was highly predictive of allergy [sensitization with vs. without allergy: Odds ratio (OR)=6.4, P=0.004; sensitization with allergy vs. controls: OR=11.5, P<0.0001]. On the contrary, IFN-gamma, IL-2, IL-4, IL-10, and IL-12 production to the ELISpot, patch test results, sensitization against other metals, and atopy score did not significantly discriminate between sensitization with and without allergy. However, IFN-gamma responses towards chromium chloride were significantly correlated with the strength of patch test reactivity (r=0.49, P=0.002). By IFN-gamma ELISpot, the average precursor cell frequency reactive to trivalent chromium could be defined as 26, 15, and 11 : 10(6) in volunteers with sensitization and allergy, with sensitization without allergy, and controls, respectively. CONCLUSIONS: In contrast to the patch test, the LTT appears to be a method that is predictive of chromium allergy. PMID- 18384429 TI - IL-13 induced increases in nitrite levels are primarily driven by increases in inducible nitric oxide synthase as compared with effects on arginases in human primary bronchial epithelial cells. AB - BACKGROUND: Exhaled nitric oxide is increased in asthma, but the mechanisms controlling its production, including the effects of T-helper type 2 (Th2) cytokines, are poorly understood. In mouse and submerged human epithelial cells, Th2 cytokines inhibit expression of inducible nitric oxide synthase (iNOS). Arginases have been proposed to contribute to asthma pathogenesis by limiting the arginine substrate available to NOS enzymes, but expression of any of these enzymes has not been extensively studied in primary human cells. OBJECTIVES: We hypothesized that primary human airway epithelial cells in air-liquid interface (ALI) culture would increase iNOS expression and activity in response to IL-13, while decreasing arginase expression. METHODS: iNOS and arginase mRNA (real-time PCR) and protein expression (Western blot and immunofluorescence) as well as iNOS activity (nitrite levels) were measured in ALI epithelial cells cultured from bronchial brushings of normal and asthmatic subjects following IL-13 stimulation. RESULTS: IL-13 up-regulated iNOS mRNA primarily at a transcriptional level in epithelial cells. iNOS protein and activity also increased, arginase1 protein expression decreased while arginase 2 expression did not change. The changes in iNOS protein correlated strongly with changes in nitrites, and inclusion of arginase (1 or 2) did not substantially change the relationship. Interestingly, iNOS mRNA and protein were not correlated. CONCLUSIONS: These results contrast with many previous results to confirm that Th2 stimuli enhance iNOS expression and activity. While arginase 1 protein decreases in response to IL-13, neither arginase appears to substantially impact nitrite levels in this system. PMID- 18384430 TI - Cow's milk allergy in adults is rare but severe: both casein and whey proteins are involved. AB - BACKGROUND: Studies on cow's milk allergy (CMA) in adults are scarce. Little is known about the clinical symptoms, eliciting doses (ED), and allergens involved. OBJECTIVE: The aim of this study was to analyse the clinical symptoms, ED and allergen recognition in adult CMA patients, compared with cow's milk (CM) sensitized, but tolerant controls. METHODS: Adult CMA patients were evaluated by standardized questionnaires (n=30), skin prick tests (SPTs) and specific IgE for CM allergens (n=18), and a double-blind placebo-controlled food challenge (DBPCFC, n=10). A control group (n=25) of CM-sensitized, but tolerant adults was included. RESULTS: The majority of CMA patients (20/30, 67%) reported severe symptoms. In all patients participating in DBPCFC, CMA was confirmed. ED for subjective symptoms (0.3-300 mg CM protein) were significantly lower than that for objective symptoms (300-9000 mg CM protein). The severity of CMA by history and ED was not correlated with SPT or IgE. Patients had higher SPT reactivity than controls for CM, alpha-lactalbumin and beta-lactoglobulin (P=0.002, P=0.014 and P=0.004) but not for casein. Specific IgE to CM tended to be higher (P=0.068) and IgE to casein was higher in patients than that in controls (P=0.016). No difference was observed for IgE to alpha-lactalbumin and beta-lactoglobulin. CONCLUSION: Adult CMA is severe in nature. ED are low, starting from 0.3 mg CM protein. Patients with CMA recognize the same major allergens (casein and whey proteins) as controls, but display a stronger SPT and IgE reactivity. PMID- 18384431 TI - Eosinophils: biological properties and role in health and disease. AB - Eosinophils are pleiotropic multifunctional leukocytes involved in initiation and propagation of diverse inflammatory responses, as well as modulators of innate and adaptive immunity. In this review, the biology of eosinophils is summarized, focusing on transcriptional regulation of eosinophil differentiation, characterization of the growing properties of eosinophil granule proteins, surface proteins and pleiotropic mediators, and molecular mechanisms of eosinophil degranulation. New views on the role of eosinophils in homeostatic function are examined, including developmental biology and innate and adaptive immunity (as well as their interaction with mast cells and T cells) and their proposed role in disease processes including infections, asthma, and gastrointestinal disorders. Finally, strategies for targeted therapeutic intervention in eosinophil-mediated mucosal diseases are conceptualized. PMID- 18384432 TI - Single-institute phase 2 study of thalidomide treatment for refractory or relapsed multiple myeloma: prognostic factors and unique toxicity profile. AB - We previously reported a pilot study of thalidomide monotherapy for Japanese patients with refractory or relapsed multiple myeloma. In the present work, we have extended this clinical trial to a single-institute phase 2 study with a larger number of patients and longer follow-up time. New information on the optimal dose and prognostic factors as well as the correlation of toxicities with treatment schedule was obtained. Fifteen of 56 (27%) patients achieved a partial response, including three cases with near-complete remission. Most patients suffered toxicities at a dose of 400 mg per day, but there was no clear dose response relationship. Thus, a lower dose such as 200 mg per day or less is considered optimal. Multivariate analyses identified only lack of response to therapy as an adverse prognostic factor for progression-free survival. Chromosomal abnormality, C-reactive protein >10 mg/L, and more than six previous courses of chemotherapy were significantly associated with shorter overall survival. Grade 3 or 4 neutropenia and thrombocytopenia were observed in 23 and 11% of patients, respectively. Grade 4 interstitial pneumonia and grade 5 pulmonary hypertension were observed; however, no patient suffered deep vein thrombosis, which has frequently been observed in other studies. Duration of therapy was closely related to the development of peripheral neuropathy. The efficacy and prognostic factors of this treatment were confirmed in long-term observation. However, special attention should be paid to toxicities such as hematological and pulmonary complications as well as peripheral neuropathy in long-term users. PMID- 18384433 TI - Vascular endothelial growth factor expression is closely related to irinotecan mediated inhibition of tumor growth and angiogenesis in neuroblastoma xenografts. AB - In the present study, irinotecan (CPT-11) was highly effective not only against the chemosensitive neuroblastoma (NB) xenografts SK-N-ASnu and TNB9, but also against the multidrug-resistant NB xenograft TS-N-2nu. SK-N-ASnu and TNB9 were significantly more responsive to low-dose daily CPT-11 treatment than to intermittent administration of one-third of the median lethal dose. For TS-N-2nu, there was no significant difference in tumor growth inhibition between the two treatment schedules. Treatment with CPT-11 alone could not completely abolish tumor growth in mice. For TNB9, tumor regrowth seemed to result from an inability to regress host vessels in the stroma during treatment and an inability to suppress host-derived vascular endothelial growth factor (VEGF) expression throughout therapy. In the multidrug-resistant TS-N-2nu, VEGF was not suppressed by low-dose therapy with CPT-11, and neurofilament-positive tumor cells escaped from apoptosis and were growth arrested at G(0)/G(1) phase. These findings suggest a mechanism for the incomplete responsiveness of TS-N-2nu to CPT-11. Our data demonstrate that diminished VEGF gene and protein expression is closely correlated with tumor growth inhibition and inhibition of angiogenesis by CPT-11 in NB xenografts. Our results further suggest that a persistent blocker of stroma derived VEGF will need to be combined with CPT-11 to completely inhibit the growth of chemosensitive NB, and that administration of CPT-11 at higher doses will be required to inhibit the growth of multidrug-resistant NB. PMID- 18384434 TI - Nuclear factor-kappaB dependency of doxorubicin sensitivity in gastric cancer cells is determined by manganese superoxide dismutase expression. AB - The role of nuclear factor-kappaB (NF-kappaB) activation in cancer cell apoptosis appears to be tailored specifically for each cell type and the type of NF-kappaB inducer. The present study aimed to determine whether or not NF-kappaB activation is associated with chemosensitivity to doxorubicin (DOX) using the DOX-sensitive SNU-601 and DOX-resistant SNU-216 gastric cancer cell lines. The effect of NF kappaB activation on DOX (1 microg/mL) sensitivity was analyzed after the suppression of NF-kappaB activation using transfection of the super-suppressive mutant form of IkappaBalpha (mIkappaBalpha) or pretreatment with pyrrolidine dithiocarbamate. In addition, the association between NF-kappaB and manganese superoxide dismutase (MnSOD) in relation to DOX sensitivity was analyzed after the modulation of MnSOD expression. The NF-kappaB activity was much higher in DOX resistant SNU-216 cells than in DOX-sensitive SNU-601 cells before and after DOX treatment. Overexpression of mIkappaBalpha or pyrrolidine dithiocarbamate pretreatment decreased the DOX resistance in SNU-601 cells with low MnSOD expression, but not in SNU-216 cells with high MnSOD expression. In comparison, the overexpression of MnSOD, which also suppressed NF-kappaB activation in both cell lines, increased DOX resistance in SNU-601 cells. Blocking of MnSOD expression using RNA interference techniques increased DOX sensitivity in SNU-216 cells, which was further augmented by the additional inhibition of NF-kappaB activity. Our results showed that whether NF-kappaB contributes to DOX sensitivity in gastric cancer cells is determined by the level of MnSOD expression. Thus, targeting both MnSOD and NF-kappaB may be helpful for increasing the efficacy of DOX treatment of DOX-resistant SNU gastric cancer cells. PMID- 18384435 TI - Genetic and epigenetic changes in aberrant crypt foci and serrated polyps. AB - Aberrant crypt foci (ACF) in colorectal mucosa are the earliest known morphological precursors to colorectal cancer and can be subclassified as dysplastic, heteroplastic (non-dysplastic), and mixed types. Serrated adenoma (SA) is a polyp with serrated architecture and dysplasia, and can be subclassified as traditional SA or sessile SA. Sessile SA is thought to be preneoplastic and differs from most lesions in the traditional SA category because of their flat morphology and general lack of cytological dysplasia. Serrated polyps include hyperplastic polyps (HP), SA, and admixed hyperplastic adenomatous polyps and are considered a morphological continuum encompassing heteroplastic ACF, HP, admixed hyperplastic-adenomatous polyps, and SA. Recent studies have uncovered other developmental pathways including a heteroplastic ACF HP/SA-carcinoma sequence and a heteroplastic ACF-adenoma-carcinoma sequence. Heteroplastic ACF histopathologically resemble HP and SA. Sporadic HP are usually present in the left colon, are small, and are considered benign. However, adenocarcinoma arising in the setting of colorectal HP or SA, especially in patients with hyperplastic polyposis, has been described. The relationship between heteroplastic ACF, HP, and colorectal cancer is less certain than that of dysplastic ACF. Here, we discuss the current understanding of genetic and epigenetic alterations in the development of colorectal cancer. Our goal is to provide a conceptual framework for understanding the heteroplastic ACF-HP/SA carcinoma sequence. PMID- 18384437 TI - Alkaline Trypan Blue as a stain for superficial fungi. PMID- 18384436 TI - Prognostic importance of T and NK-cells in a consecutive series of newly diagnosed patients with chronic lymphocytic leukaemia. AB - Patients with chronic lymphocytic leukaemia (CLL) have a variable clinical course. The identification of modifiable characteristics related to CLL-specific survival may provide opportunities for therapeutic intervention. The absolute number of T-cell and natural killer (NK)-cells was calculated for 166 consecutive patients with CLL evaluated by flow cytometry at Mayo Clinic < or = 2 months of diagnosis. The size of the T-cell/NK-cell compartment relative to the size of the malignant monoclonal B-cell (MBC) compartment was evaluated by calculating NK:MBC and T:MBC ratios. Patients exhibited substantial variation in the absolute number of T- and NK-cells as well as T:MBC and NK:MBC ratios at diagnosis. Higher T:MBC and NK:MBC ratios were observed among patients with early stage and mutated IGHV genes (all P < or = 0.0003). As continuous variables, both T:MBC ratio (P-value = 0.03) and NK:MBC ratio (P-value = 0.02) were associated with time to treatment (TTT). On multivariate Cox modelling including stage, CD38, absolute MBC count, NK:MBC ratio and T:MBC ratio, the independent predictors of TTT were stage, T:MBC ratio and NK:MBC ratio. These findings suggest that measurable characteristics of the host immune system relate to the rate of disease progression in patients with newly diagnosed CLL. These characteristics can be modified and continued evaluation of immunomodulatory drugs, vaccination strategies and cellular therapies to delay/prevent disease progression are warranted. PMID- 18384438 TI - Subcellular activation site of caspase-3 in apoptotic keratinocytes observed in lichenoid tissue reaction. PMID- 18384439 TI - CD26/dipeptidyl-peptidase IV in psoriatic skin: upregulation and topographical changes. AB - BACKGROUND: Psoriasis is known to affect 2-3% of the population and can be considered an organ-specific autoimmune disease. CD26/dipeptidyl-peptidase IV (DPP-IV) is a membrane-bound protease with diverse properties. In theory, the expression of CD26/DPP-IV has common grounds with three principal key players of the psoriatic pathogenesis: keratinocytes, T cells and cytokines. OBJECTIVES: To assess CD26/DPP-IV expression in psoriasis in order to expand on the search for complementary biomarkers related to inflammation and proliferation in psoriasis. METHODS: The pattern of expression of CD26/DPP-IV was investigated on the mRNA-, protein- and enzyme-functionality level using immunohistochemical, immunofluorescent and enzyme activity labelling techniques. RESULTS: An 11-fold significant increase of CD26/DPP-IV on the mRNA level was demonstrated in psoriatic epidermal sheets compared with normal skin. Immunohistochemistry on psoriatic sections showed a distinct patchy honeycomb-like CD26/DPP-IV staining in the suprapapillary layers. Moreover, a clearly distinguishable column-like staining pattern throughout the suprabasal compartment along the rete ridges was seen, whereas in normal skin these patterns were absent. Strikingly, CD26/DPP-IV enzyme activity correlated with this immunohistochemical reactivity pattern for the CD26/DPP-IV protein. The T-cell bound expression of CD26/DPP-IV in psoriatic skin was explicitly present, albeit in small quantities. CONCLUSIONS: Our data provide clear evidence for a versatile upregulation of CD26/DPP-IV expression in psoriatic (epi)dermis. Although the exact functional contribution remains speculative, the topographical distribution of this complex multifunctional protein suggests a suitable role as a complementary biomarker in psoriasis. PMID- 18384440 TI - Psoriasis treatment with etanercept and efalizumab: clinical strategies influencing treatment outcome. AB - BACKGROUND: Multiple trials have been conducted in which the safety and efficacy of different biological therapies for psoriasis have been studied. However, the treatment course in clinical practice is different from the setting in which trials are conducted. OBJECTIVES: The evaluation of the efficacy, safety and adverse events of etanercept and efalizumab treatment in daily practice and the investigation of interfering clinical strategies that could be of influence on treatment outcome. METHODS: A prospective cohort consisting of 101 patients with high-need psoriasis was followed for 2 years and analysed. Patients were treated with etanercept or efalizumab between February 2005 and May 2007. Efficacy, safety and adverse events were investigated. Furthermore, all accompanying factors of which an influence on treatment efficacy outcome was suspected were registered, including treatment interruptions, dosage adjustments and combinations of therapies. RESULTS: Etanercept and efalizumab treatment was effective and safe in most patients. However, in many cases the treatment course was characterized by unsatisfactory efficacy (83%), necessitating combination therapies or dosage adjustments. Treatment interruptions occurred in 56% (etanercept 2x50 mg group), 84% (etanercept 2x25 mg group) and 10% (efalizumab treated patients). CONCLUSIONS: Treatment of high-need psoriasis patients in daily practice is highly different from treatment courses in clinical trials. Frequently applied clinical strategies such as treatment interruptions, dosage adjustments and combinations of treatments influence treatment outcome in routine treatment in comparison with randomized controlled trials. Information about treatment with these new drugs in daily clinical practice is important for adjusting treatment schedules and guidelines. PMID- 18384441 TI - Nail and periungual toxicity following capecitabine therapy. PMID- 18384442 TI - Evidence of the in vivo esterification of budesonide in human airways. AB - AIMS: Budesonide, unlike fluticasone propionate, undergoes fatty acid esterification in the lungs, and there is a need to characterize fully the distribution and fate of the two drugs after inhalation in humans. METHODS: This open-label, randomized study was performed in adults undergoing whole lung or lobar resection resulting from lung cancer. Patients were given single 1000-mug doses of both budesonide and fluticasone propionate via dry powder inhalers before surgery. Tissue samples from peripheral and central lung, an ex vivo bronchial brush sample and intercostal muscle, together with plasma samples, were taken during surgery and analysed by liquid chromatography plus tandem mass spectrometry. RESULTS: Lung tissue samples were obtained from 22 patients at surgery, 1-43 h after drug dosing. Budesonide was detectable from earliest sampling in central and peripheral lung tissue up to 10 h (in six of 22 samples), fluticasone propionate up to 22 h after inhalation (in 16 of 22 samples), and budesonide oleate up to 43 h after inhalation (in 21 of 22 samples). Budesonide, but not fluticasone propionate, was detected in intercostal muscle for up to 10 h after inhalation. Bronchial brush samples showed the presence of fluticasone propionate for up to 18 h, suggesting the presence of undissolved drug powder particles in the airway lumen. CONCLUSION: Sustained retention of esterified budesonide in the lungs supports the prolonged duration of action of budesonide and suitability for once-daily administration. PMID- 18384443 TI - Effects of imatinib (Glivec) on the pharmacokinetics of metoprolol, a CYP2D6 substrate, in Chinese patients with chronic myelogenous leukaemia. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: Imatinib, a tyrosine kinase inhibitor, exhibits a competitive inhibition on the CYP450 2D6 isozyme with a K(i) value of 7.5 microm. However, the clinical significance of the inhibition and its relevance to 2D6 polymorphisms have not been evaluated. The pharmacokinetics of imatinib have been well studied in Caucasians, but not in a Chinese population. Metoprolol, a CYP2D6 substrate, has different clearances among patients with different CYP2D6 genotypes. It is often used as a CYP2D6 probe substrate for clinical drug-drug interaction studies. WHAT THIS STUDY ADDS: Co-administration of imatinib at 400 mg twice daily increased the plasma AUC of metoprolol by approximately 23% in 20 Chinese patients with chronic myeloid leukaemia (CML), about 17% increase in CYP2D6 intermediate metabolizers (IMs) (n = 6), 24% in extensive metabolizers (EMs) (n = 13), and 28% for the subject with unknown 2D6 status (n = 1) suggesting that imatinib has a weak to moderate inhibition on CYP2D6 in vivo. * The clearance of imatinib in Chinese patients with CML showed no difference between CYP2D6 IMs and EMs, and no major difference from Caucasian patients with CML based on data reported in the literature. AIMS To investigate the effect of imatinib on the pharmacokinetics of a CYP2D6 substrate, metoprolol, in patients with chronic myeloid leukaemia (CML). The pharmacokinetics of imatinib were also studied in these patients. METHODS: Patients (n = 20) received a single oral dose of metoprolol 100 mg on day 1 after an overnight fast. On days 2-10, imatinib 400 mg was administered twice daily. On day 8, another 100 mg dose of metoprolol was administered 1 h after the morning dose of imatinib 400 mg. Blood samples for metoprolol and alpha-hydroxymetoprolol measurement were taken on study days 1 and 8, and on day 8 for imatinib. RESULTS: Of the 20 patients enrolled, six patients (30%) were CYP2D6 intermediate metabolizers (IMs), 13 (65%) extensive metabolizers (EMs), and the CYP2D6 status in one patient was unknown. In the presence of 400 mg twice daily imatinib, the mean metoprolol AUC was increased by 17% in IMs (from 1190 to 1390 ng ml(-1) h), and 24% in EMs (from 660 to 818 ng ml(-1) h). Patients classified as CYP2D6 IMs had an approximately 1.8-fold higher plasma metoprolol exposure than those classified as EMs. The oral clearance of imatinib was 11.0 +/- 2.0 l h(-1) and 11.8 +/- 4.1 l h(-1) for CYP2D6 IMs and EMs, respectively. CONCLUSIONS: Co-administration of a high dose of imatinib resulted in a small or moderate increase in metoprolol plasma exposure in all patients regardless of CYP2D6 status. The clearance of imatinib showed no difference between CYP2D6 IMs and EMs. PMID- 18384444 TI - Thrombolysis or nothing for acute myocardial infarction? It's all the same! AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: The methodological and ethical value of equivalence and non-inferiority trials is questioned. These studies are still increasingly used in drug evaluation and accepted by the scientific community and the regulatory authorities. WHAT THIS STUDY ADDS: By applying the hypothesis of an equivalence trial of saruplase and streptokinase (SK) we proved that the no thrombolysis approach in the GISSI trial was equivalent to thrombolysis with SK, i.e. death rates in patients given SK or not were similar enough to consider the no-thrombolysis regimen equivalent to thrombolytic treatment. These data illustrate the unreliability of equivalence trials, which can even disprove consolidated clinical evidence such as the efficacy of thrombolysis in acute myocardial infarction. Equivalence trials should not be considered an option by the scientific community and should not be accepted as a basis for marketing authorization by the regulatory authorities. AIMS To assess the reliability of equivalence trials we tested whether the no-thrombolysis approach was equivalent to thrombolysis with streptokinase (SK) in acute myocardial infarction. METHODS: We applied the hypothesis of an equivalence trial of a recombinant plasminogen activator and SK to the GISSI-1 control group. RESULTS: In at least one of three subsets randomly extracted from the GISSI database the equivalence criterion was satisfied, i.e. death rates in patients given SK or not were similar enough to consider the no-thrombolysis regimen equivalent to thrombolytic treatment. Two thirds of 100 replications of the sampling gave this result. CONCLUSIONS: These findings suggest the unreliability of equivalence trials, which should neither be adopted by the scientific community nor accepted by the regulatory authorities. PMID- 18384445 TI - Estimation of the impact of noncompliance on pharmacokinetics: an analysis of the influence of dosing regimens. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: Patient compliance is better with formulations that require less frequent dosing than with formulations that require more frequent dosing. Intramuscular risperidone and long-acting oxybutynin are two examples of medicines reformulated for less frequent dosing. However, it is not clear whether better compliance with less frequent dosing regimens translates to improved therapeutic outcome. WHAT THIS STUDY ADDS: At equivalent daily doses and typical patterns of compliance, fortnightly intramuscular depot administrations of risperidone provide better pharmacokinetic coverage than once-daily oral dosing. Once-daily dosing of oxybutynin is no better at maintaining pharmacokinetic exposure than twice-daily dosing at half strength. The use of simulated compliance data as input to pharmacokinetic models is useful to assess the impact of noncompliance on internal drug exposure. AIMS: To determine whether, for oxybutynin and risperidone, drug exposure is better with less frequent dosing regimens than with regimens that require more frequent dosing. METHODS: Pharmacokinetic models of oxybutynin (5 mg twice-daily and 10 mg once-daily) and risperidone (2 mg once-daily orally and 25 mg fortnightly intramuscular injection) were developed. Simulations of multiple doses were performed by use of stochastic models of dose-taking compliance and clinic visit attendance. RESULTS: At therapeutic concentrations and with typical patterns of noncompliance, intramuscular injections of risperidone resulted in a 41% (SD 12%) greater pharmacokinetic coverage than the oral dose, 76% (SD 10%) vs. 35% (SD 7%). No discernable differences were evident between once- and twice-daily formulations of oxybutynin, 29.2% (SD 10%) vs. 29.0% (SD 13%). CONCLUSIONS: For equivalent doses for each drug, the longer acting preparation of risperidone, but not oxybutynin, is pharmacokinetically more forgiving of noncompliance than the shorter acting counterparts. Further analysis is required to confirm whether these observations are valid clinically. PMID- 18384446 TI - Selective COX-2 inhibitors, NSAIDs and congestive heart failure: differences between new and recurrent cases. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: Pharmaco-epidemiological studies have shown that in susceptible individuals, nonsteroidal anti-inflammatory drugs (NSAIDs) and selective cyclooxygenase (COX)-2 inhibitors increase the risk of developing congestive heart failure (CHF). Recently published studies have found lower relative risk (RR) estimates than the initial studies published in 1998 2000. It is unclear whether the level of risk is elevated equally in first time and recurrent cases of CHF. WHAT THIS STUDY ADDS: This study found low-level, statistically nonsignificant elevations of risk with NSAIDs and COX-2 inhibitors. There was a much higher level of recent use of NSAIDs/COX-2 inhibitors among first-time cases than among recurrent cases of CHF. * The dilution of the RR over successive studies, and the differences between first-time and recurrent cases noted here, suggest that prescribing doctors have heeded advice about the cardiovascular risks of NSAIDs and extended this practice to selective COX-2 inhibitors. AIMS: To quantify the association between treatment with nonsteroidal anti-inflammatory drugs (NSAIDs) and selective cyclooxygenase (COX)-2 inhibitors and hospitalization due to congestive heart failure (CHF); to determine if the risk varies between first and subsequent episodes of CHF. METHODS: We conducted a case-control study of the relationship between recent use of NSAIDs and COX-2 inhibitors and hospitalization with CHF. Cases (n = 530) were patients admitted to hospital with a primary diagnosis of CHF. Controls (n = 1054) were subjects without CHF who were admitted to the same hospitals as the cases. They were frequency matched to cases on the basis of age and sex. Structured interviews were used to obtain information on a number of study factors, including recent use of NSAIDs and COX-2 inhibitors. Relative risks (RRs) were estimated from exposure odds ratios, adjusted for a range of potential confounders. RESULTS: Overall, NSAIDs and COX-2 inhibitors had been taken by 249 (23.6%) controls in the week before admission to hospital. Use of any NSAID/COX-2 inhibitor was recorded in 81/285 (28.4%) first-time cases compared with 38/245 (15.5%) in recurrent cases: difference 12.9% (95% confidence interval 5.9, 19.8) (P = 0.0004). The adjusted RRs for first hospital admission for CHF with different drug exposures were: NSAIDs 1.1 (0.67, 1.83), rofecoxib 1.29 (0.78, 2.13) and celecoxib 1.47 (0.85, 2.53). CONCLUSIONS: We found weak and statistically nonsignificant associations between use of NSAIDs and COX-2 inhibitors and hospitalization with CHF. This low RR is consistent with the results of recently published studies, but not with early studies that found an approximate doubling of risk with use of NSAIDs. The dilution of risk and the significantly lower levels of prescribing for recurrent than for first-time cases of heart failure suggest that prescribing doctors heeded messages that NSAIDs may precipitate CHF in vulnerable individuals, and that they have applied the same message to selective COX-2 inhibitors. PMID- 18384448 TI - Incidence, clinical findings and management of intraoperative floppy iris syndrome associated with tamsulosin. AB - PURPOSE: To determine the risk ratios and incidence of intraoperative floppy iris syndrome (IFIS) during cataract surgery in patients using tamsulosin, and to assess management strategies for IFIS. METHODS: We performed a non-randomized, observational, prospective study, in which 594 eyes of 579 patients undergoing cataract surgery were enrolled. Surgeons were masked to the patients' drug history. Usage or non-usage of tamsulosin, duration of tamsulosin use, presence or absence of IFIS, management of IFIS and intraoperative complications were recorded in the patients' theatre notes. RESULTS: Twelve of 15 (80%) IFIS patients were taking systemic tamsulosin. Twelve of 23 (52%) patients using tamsulosin showed features of IFIS. The odds ratios (ORs) and relative risk (RR) ratios show strong positive correlations between tamsulosin use and IFIS. The ORs and RR ratios and the 95% confidence intervals (CIs) are as follows: OR 206.5 (95% CI 50.9-836.5); RR 99.3 (95% CI 30.0-327.8). There were no statistically significant differences between patients with or without IFIS, who were using tamsulosin, in terms of age or duration of tamsulosin use (p > 0.05). Seven eyes (46.6%) with IFIS were successfully managed with epinephrine. Eight eyes (53.4%) with IFIS needed iris hooks. CONCLUSIONS: Patients using tamsulosin appear to be at high risk of IFIS during cataract surgery. The occurrence of IFIS may not be affected by duration of tamsulosin use or age. Epinephrine may be effective in approximately 50% of eyes with IFIS. The iris hook procedure represents an effective management strategy in IFIS. PMID- 18384449 TI - Association between TNFA-308 G/A polymorphism and sensitization to para phenylenediamine: a case-control study. AB - BACKGROUND: Para-phenylenediamine (PPD) and related chemicals are common contact sensitizers, frequently causing allergic contact dermatitis (ACD). The cytokine tumor necrosis factor-alpha (TNF-alpha) plays a key role in contact sensitization. METHODS: In this case-control study, we evaluated the distribution of variations in the regulatory region of the gene for TNF-alpha (TNFA-308 G/A) in 181 Caucasian individuals with a history of ACD and sensitization to PPD and 161 individuals with no history of sensitization to PPD. RESULTS: The frequency of GA or AA TNFA genotypes was significantly higher in individuals sensitized to PPD than in age- and gender-matched controls giving an odds ratio (OR) of 2.16 (95% confidence interval, CI: 1.35-3.47; P = 0.0016). This relation was even more pronounced when restricting cases to females over 45 years (OR = 3.71; 95% CI: 1.65-8.31; P = 0.0017) vs younger females (less than or equal to 45 years; OR = 2.41; 95% CI: 1.03-5.65; P = 0.044) or males (OR = 1.05; 95% CI: 0.449-2.47; P = 1.0). In addition, a logistic regression model revealed a significant effect for TNFA-308 AA and AG vs GG genotype (point estimate = 2.152; 95% Wald CI: 1.332 3.477). CONCLUSIONS: These findings suggest a possible role for the TNFA-308 genetic polymorphism as a susceptibility factor for chemically induced ACD. PMID- 18384450 TI - Occupational allergy in a researcher due to Ole e 9, an allergenic 1,3-beta glucanase from olive pollen. PMID- 18384451 TI - Prevalence of asthma in a large group of Israeli adolescents: influence of country of birth and age at migration. AB - BACKGROUND: The occurrence of asthma has geographic variations and is lower in developing compared with industrialized countries. Both environmental and genetic factors may influence its prevalence. We aimed to evaluate the importance and effect of immigration (country of birth and age at immigration to Israel) on the prevalence of asthma in a large group of Israeli adolescents. METHODS: Computerized medical records of 17-year-old adolescents, who underwent routine examination before military recruitment, were studied. The sample comprised both native-born Israelis (NBI) and immigrants from Ethiopia, the Former Soviet Union (FSU), and Western countries (WC). Asthma was defined as clinical symptoms and signs compatible with the disease accompanied by abnormal spirometry or documented chronic use of inhaled steroids. RESULTS: Our cohort consisted of 1 466 654 adolescents, including 1 317 556 (89.8%) NBI and 149 098 (10.2%) immigrants. The prevalence of asthma at age 17 was higher in NBI compared with Ethiopian immigrants [4.7% (61 921) vs 2.6% (418), respectively, P < 0.0005], lower compared with immigrants from WC [5.6% (2177), P < 0.0005], and similar to immigrants from the FSU. Further analysis of the association between age at immigration and the risk for developing asthma showed that the younger immigrants from the FSU and Ethiopia arrived to Israel, the higher their prevalence of asthma at the age of 17 was. CONCLUSIONS: Both environmental and genetic factors seem to influence the prevalence of asthma in 17-year-old adolescents. However, the higher risk for developing asthma associated with young age of immigration points toward an environmental predominance. PMID- 18384452 TI - Cytokine and chemokine expression in the skin from patients with maculopapular exanthema to drugs. AB - BACKGROUND: Maculopapular exanthema (MPE) is the most frequent clinical manifestation of nonimmediate allergic reactions to drugs and T helper 1 (Th1) cytokines and CD4(+) T cells have been shown to play an important role in its pathogenesis. We assessed the role of cytokines and chemokines and their receptors in the pathogenesis of MPE. METHODS: We evaluated skin biopsies and peripheral CD4(+) and CD8(+) T cells from 27 patients during the acute phase of the reaction and 26 exposed controls. Semiquantitative real-time PCR was performed to determine the expression of cytokines and chemokines and their receptors and immunohistochemistry was used to determine the same chemokines and their receptor proteins in skin. RESULTS: There was a high expression of the Th1 cytokines interferon-gamma (P = 0.006) and tumor necrosis factor-alpha (P = 0.022) in skin and CD4(+) T cells (P = 0.007 and P = 0.005, respectively); and of the Th1 chemokines CXCL9 (P = 0.005) and CXCL10 (P = 0.028) in the skin, while their receptor CXCR3 was increased in skin (P = 0.006) and CD4(+) T cells (P = 0.03). Homing chemokine receptors were also increased: CCR6 in skin (P = 0.026) and CD4(+) T cells (P = 0.016), and CCR10 only in CD4(+) T cells (P = 0.016), as well as their ligands, CCL20 and CCL27, in skin alone. Immunohistochemistry confirmed these results. CONCLUSIONS: These data show significant differences in the expression of chemokines and chemokine receptors, related with a Th1 profile, in both skin biopsies and peripheral CD4(+) T cells in patients with drug-induced MPE. PMID- 18384453 TI - Psychiatric disturbances in CADASIL: a brief review. AB - BACKGROUND: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited cerebrovascular disease, clinically characterized by a variable combination of migraine, recurrent transient ischemic attack (TIA) or lacunar strokes, cognitive decline, and mood disturbances. However, the assessment of psychiatric disturbances in this disease has never been carried out systematically. METHODS: This paper presents a brief review of the literature regarding the occurrence of psychiatric disorders in CADASIL patients. RESULTS: The prevalence of psychiatric disorders in CADASIL patients is reported to range from 20% to 41%. The psychiatric disturbances reported with the highest frequency are mood disturbances (9-41%). Pooling together the studies and considering a total of 454 CADASIL patients reported in the literature, 106 of these were affected by mood disturbances (24%). The majority of studies however did not use any defined criteria to assess the presence of psychiatric disorders and diagnoses were mainly based on history or review of clinical records. CONCLUSIONS: The review of CADASIL literature suggests the need for a more structured approach to the investigation of these disturbances that are highly prevalent and may greatly impact quality of life in these patients. PMID- 18384454 TI - In-hospital direct cost of acute ischemic and hemorrhagic stroke in Greece. AB - OBJECTIVES: The geographic inequity and the wide variation in the patterns of care of stroke found across Europe together with the lack of health economics evaluation in Greece led to this prospective study, aiming to provide data on in hospital direct cost of patients with an acute stroke in Greece, and to identify independent prognostic factors. METHODS: Demographic and clinical data were recorded on 429 consecutive patients with an acute ischemic or hemorrhagic stroke admitted to a tertiary care hospital in Greece during a period of 18 months. The costs incurred were estimated using the official financial charts listing in euro (euro), the real expenditure of all hospital departments. RESULTS: The direct in hospital cost for all stroke cases was 1,551,445euro for a total of 4674 days (331.9euro per day in-hospital). The mean in-hospital cost per stroke patient was 3624.9euro (+/-2695.4). Hemorrhagic strokes were significantly more expensive than the ischemic strokes [mean 5305.4 (+/-4204.8)euro and 3214.5 (+/ 1976.2)euro, respectively) and lacunar strokes the least expensive among ischemic stroke subtypes. The length of stay was highly correlated with in-hospital total cost. Multivariate linear regression analysis showed that admission ward, stroke severity on admission, stroke type and status discharge were independent predictors of cost. CONCLUSIONS: Purchasers in our health services should differentiate in their cost estimates and pricing schemes between types of cerebrovascular events. Future studies should focus on modifiable factors related, not only with stroke characteristics, but also with operational policies of hospitals, that may influence length of stay. PMID- 18384455 TI - Enhanced plasma levels of LIGHT in patients with acute atherothrombotic stroke. AB - OBJECTIVES: As a member of the tumor necrosis factor superfamily (TNFSF), LIGHT (TNFSF14) was recently found to be associated with platelets and released upon activation. Increased plasma levels of LIGHT have been reported in patients with myocardial infarction and unstable angina. The aim of the study was to analyze plasma levels of LIGHT in acute ischemic atherosclerotic stroke. MATERIALS AND METHODS: The soluble LIGHT protein was analyzed by an enzyme-linked immunosorbent assay in peripheral blood of patients with acute ischemic atherosclerotic stroke (n = 20), asymptomatic carotid stenosis (n = 19) and normal controls (n = 23). RESULTS: During the initial 24 h after onset, the stroke patients had an increased plasma LIGHT levels as compared with normal controls. Moreover, the plasma LIGHT levels of the stroke patients were correlated with blood platelet count (r = 0.6341, P = 0.0027). CONCLUSION: The elevated LIGHT levels may reflect a persistent chronic inflammatory response that may have been induced during early stages of the disease. We speculate that this derangement of LIGHT may be important for atherogenetic process of ischemic stroke. PMID- 18384456 TI - Benzodiazepines in epilepsy: pharmacology and pharmacokinetics. AB - Benzodiazepines (BZDs) remain important agents in the management of epilepsy. They are drugs of first choice for status epilepticus and seizures associated with post-anoxic insult and are also frequently used in the treatment of febrile, acute repetitive and alcohol withdrawal seizures. Clinical advantages of these drugs include rapid onset of action, high efficacy rates and minimal toxicity. Benzodiazepines are used in a variety of clinical situations because they have a broad spectrum of clinical activity and can be administered via several routes. Potential shortcomings of BZDs include tolerance, withdrawal symptoms, adverse events, such as cognitive impairment and sedation, and drug interactions. Benzodiazepines differ in their pharmacologic effects and pharmacokinetic profiles, which dictate how the drugs are used. Among the approximately 35 BZDs available, a select few are used for the management of seizures and epilepsy: clobazam, clonazepam, clorazepate, diazepam, lorazepam and midazolam. Among these BZDs, clorazepate has a unique profile that includes a long half-life of its active metabolite and slow onset of tolerance. Additionally, the pharmacokinetic characteristics of clorazepate (particularly the sustained-release formulation) could theoretically help minimize adverse events. However, larger, controlled studies of clorazepate are needed to further examine its role in the treatment of patients with epilepsy. PMID- 18384457 TI - Pitfalls in diagnosing limbic encephalitis - a case report. AB - BACKGROUND: The syndrome of limbic encephalitis (LE) is characterized by subacute onset of temporal lobe epilepsy, loss of short-term memory, cognitive confusion and psychiatric symptoms. AIM: We report a patient with pharmacoresistant epilepsy who underwent presurgical video-electroencephalogram (EEG)-monitoring with normal psychiatric and neuropsychological findings. METHODS: Magnetic resonance imaging (MRI) revealed a hyperintense lesion within the right amygdala but no contrast enhancement. Analysis of cerebrospinal fluid (CSF) showed pleocytosis and positive oligoclonal bands, but all tests for neurotropic viruses or borrelia antibodies were negative. Presurgical evaluation identified a right mesiotemporal focus. RESULTS: As a tumour was the most likely differential diagnosis, we performed selective amygdalohippocampectomy of the right hemisphere. Subsequent histopathological examination revealed the surprising diagnosis of LE. As a consequence, tumour screening was initiated and a testicular carcinoma with high anti-Ma2-antibody titres was detected. Following surgical and chemotherapeutical treatment, the patient was seizure-free and Ma2 antibodies decreased below detection limits. Conclusion - This case report highlights that LE has to be considered even in patients with atypical clinical presentation, i.e. without neuropsychological deficits, if CSF analysis reveals an inflammatory response. When LE is diagnosed, extensive tumour search is mandatory to detect and treat the paraneoplastic origin of LE. Therapeutic strategies of LE include surgical treatment as well as early immunosuppression. PMID- 18384458 TI - Epilepsy outcomes in elderly treated with topiramate. AB - OBJECTIVES: To explore effectiveness, tolerability and quality of life in elderly patients with epilepsy treated with topiramate. METHODS: One year, open-label, flexible-dosing clinical trial. RESULTS: One hundred and seven patients (mean age 69 years, 53% men) were studied during 273 +/- 141 days. The average final dose in monotherapy was 98 mg/day vs 153 mg/day in adjunctive treatment. Mean monthly cumulative seizure frequency decreased from 3.7 +/- 15 to 1.6 +/- 7.7 (n = 101, P < 0.0001), 78% of patients with seizures at baseline (n = 102) achieved at least 50% reduction in seizure frequency, 44% were seizure-free throughout the trial. Total scores on the quality of life in epilepsy inventory (QOLIE-31) improved from 57 +/- 17 to 68 +/- 18 (n = 64, P < 0.0001). The most frequently reported adverse events included convulsions, dizziness and tiredness. CONCLUSIONS: Elderly patients treated with topiramate showed marked reductions in seizures, good tolerability and significant improvements in several aspects of quality of life. PMID- 18384459 TI - Brain magnetic resonance imaging abnormalities in neuromyelitis optica. AB - OBJECTIVE: Brain abnormalities in neuromyelitis optica (NMO) attracted much attention. Our study was to identify the brain magnetic resonance imaging (MRI) abnormalities in Chinese NMO patients. METHODS: Patients who fulfilled the latest diagnostic criteria of NMO proposed by Wingerchuk et al. [Neurology 66 (2006) 1485] and whose brain MRI did not meet the multiple sclerosis (MS) criteria of McDonald et al. [Ann Neurol 50 (2001) 121] were selected to perform MRI scanning of the brain, spinal cord and optic nerves. RESULTS: Twenty-eight of 33 patients (84.8%) had abnormal MRI findings. Twenty-two patients (66.7%) presented with well-defined brain parenchymal lesions and the other six patients (18.2%) with macroscopic symmetrical diffuse hyperintensities in deep white matter. Fifteen of 22 patients had more than one lesion (> or =2 lesions) and the other seven patients had single lesion. In the supratentorium, most lesions were punctate or small round dot and non-specific in juxtacortical, subcortical and deep white matter regions, a few were patchy atypical confluent lesions. Brainstem was easily involved (14/33, 42.4%) especially in medulla (7/33, 21.2%). CONCLUSIONS: This study demonstrates the characteristics of brain MRI abnormalities in Chinese NMO patients, which are helpful to the revision of diagnostic criteria for NMO. PMID- 18384460 TI - Effects of exercise on the morphology of the myenteric neurons of the duodenum of Wistar rats during the ageing process. AB - This study aimed to evaluate the effects of regular physical activity on the morphology of the myenteric plexus of the duodenum in rats during the ageing process. To this end, 45 Wistar rats were divided into three groups: C (sedentary - 6 months old), S (sedentary - 12 months old) and T (trained - 12 months old). The animals of group S were given with a physical activity programme consisting of a 10-min-treadmill workout once a week. The animals of group T were submitted to the physical activity programme five times a week. Their duodenums were collected and submitted to the techniques of nicotinamide adenine dinucleotide (NADH)-diaphorase enzyme histochemistry for whole-mount preparations and transmission electron microscopy. No differences in the constitution of the myenteric plexuses were found when the sedentary and trained groups were compared with the control group. The ultrastructural features were similar for the three groups. However, it was verified that the physical activity of the trained animals resulted in a similar myenteric neuron morphology to that of the adult animals (6 months old), thereby confirming its beneficial effect, as the sedentary animals had larger alterations in the collagen fibrils and the basal membrane that occur through ageing. The quantitative analysis showed that the NADH-diaphorase positive neurons decreased with ageing and increased with physical activity (P > 0.05). No significant alteration (P > 0.05) in the neuronal profile area of the NADH-diaphorase positive neurons has been observed with ageing. PMID- 18384461 TI - Lack of association between CAST SNPs and meat tenderness in sheep. PMID- 18384463 TI - Microsatellite flanking regions: a SNP mine in South African abalone (Haliotis midae). PMID- 18384462 TI - Chromosomal assignment of eight porcine genes involved in apoptosis. PMID- 18384464 TI - Characterization of swine adiponectin and adiponectin receptor polymorphisms and their association with reproductive traits. AB - In this study, polymorphisms in genes encoding porcine adiponectin (ADIPOQ) and its receptors (ADIPOR1 and ADIPOR2) were evaluated for associations with reproductive traits in a Landrace sow population. Sixteen SNPs were identified, and among these, associations were found between reproductive traits and five SNPs. Heterozygous multiparous females for SNP ADIPOQEF601160:c.178G>A had fewer stillborn piglets (P < 0.05) and shorter weaning-to-oestrus intervals (P < 0.05). Multiparous females bearing the mutant allele for SNP ADIPOQEF601160:c.*1094_1095insC gave birth to fewer stillborn piglets (P < 0.05). In addition, selection for the ADIPOQ [A;C] haplotype is expected to result in multiparous sows having the lowest number of stillborn piglets and shorter weaning-to-oestrus intervals. In second-parity sows, the polymorphism in ADIPOR1 (AY856513:c.*129A>C) showed significant associations with live-born (P < 0.01) and stillborn (P < 0.05) piglets. In multiparous sows, a significant association was observed for an ADIPOR2 polymorphism (AY856514:c.*112G>A), with the c.*112GA genotype associated with shorter weaning-to-oestrus intervals (P < 0.01). Haplotype analyses of ADIPOR2 SNPs revealed that selection in favour of the [A;C] haplotype and against the [G;G] haplotype may result in sows having an increased number of live-born piglets and shorter weaning-to-oestrus intervals. We have therefore described specific SNPs and haplotypes that are associated with large litter size, fewer stillborn and mummified piglets and shorter weaning-to-oestrus intervals. Selection for these SNPs and haplotypes is a strategy to improve reproductive success in pigs. PMID- 18384465 TI - Differences in the expression of the ASIP gene are involved in the recessive black coat colour pattern in sheep: evidence from the rare Xalda sheep breed. AB - Here we have tested the hypothesis of association between different levels of agouti signalling peptide (ASIP) mRNA and the recessive black coat colour in the rare Xalda breed of sheep. To deal with this task, we first tested the possible action of both the dominant black extension allele (E(D)) and a 5-bp deletion (X99692:c.100_104del; A(del)) in the ovine ASIP coding sequence on the black coat colour pattern in 188 Xalda individuals. The E(D) allele was not present in the sample and only 11 individuals were homozygous for the A(del)ASIP allele. All Xalda individuals carrying the A(del)/A(del) genotype were phenotypically black. However, most black-coated individuals (109 out of 120) were not homozygous for the 5-bp deletion, thus rejecting the A(del)/A(del) genotype as the sole cause of recessive black coat colour in sheep. Differences in expression of ASIP mRNA were assessed via RT-PCR in 14 black-coated and 10 white-coated Xalda individuals showing different ASIP genotypes (A(wt)/A(wt), A(wt)/A(del) and A(del)/A(del)). Levels of expression in black animals were significantly (P < 0.0001) lower than those assessed for white-coated individuals. However, the ASIP genotype did not influence the ASIP mRNA level of expression. The consistency of these findings with those recently reported in humans is discussed, and the need to isolate the promoter region of ovine ASIP to obtain further evidence for a role of ASIP in recessive black ovine pigmentation is pointed out. PMID- 18384466 TI - Acute and continuation therapy in unipolar depression: observations from the run in phase of a maintenance trial. AB - OBJECTIVE: The aim of the study was to analyze treatments and outcome in depressed patients. METHOD: Patients with recurrent depressive disorder (n = 289), recruited for a prophylaxis study, were followed up in hospital settings for 6 months with diagnostic and depression ratings at baseline and monthly depression ratings. Data on psychotropic drugs were retrieved from hospital case records. Independent associations between baseline, treatment and outcome variables were examined by logistic regression models. RESULTS: Depressive symptoms subsided gradually. After 6 months, 21% had dropped out, 43% were rated as remitted (HAM-D-17 <8) and 8% had not responded (HAM-D-17 >15). Patients once remitted rarely relapsed (<5%). All patients received antidepressant drugs, half of them more than one (2-4) as well as other psychotropic drugs. Patients responding poorly received more frequently multiple antidepressants, tricyclic antidepressants, hypnosedatives, lithium and/or antipsychotics. CONCLUSION: The 6 month outcome was generally poor. Choice of treatment appeared at least partly to be determined by the therapeutic outcome. PMID- 18384467 TI - Antidepressant treatment is associated with a reduction in suicidal ideation and suicide attempts. AB - OBJECTIVE: To measure changes in suicidal behaviours during 6 months of treatment with antidepressants. METHOD: A group of depressed patients (n = 195) were assessed for suicidal behaviours in the 6 months prior to treatment. They were prospectively assessed for suicidal behaviours during 6 months of treatment with antidepressants. RESULTS: Patients who made suicide attempts fell from 39 in the 6 months prior to treatment to 20 during treatment. Significant suicidal ideation reduced from 47% at baseline to 14% at 3 weeks remaining below this during the rest of the treatment. Twenty patients had emergent suicidal ideation; five of them had not experienced some level of suicidal behaviour in the 6 months prior to treatment. CONCLUSION: Suicide behaviours are common in depressed out patients. Antidepressant treatment is associated with a rapid and significant reduction in suicidal behaviours. The rate of emergent suicidal behaviour was low and the risk benefit ratio for antidepressants appears to favour their use. PMID- 18384468 TI - The significance of complete interruption of large spontaneous portosystemic collaterals in adult living donor liver transplantation as a graft salvage procedure. PMID- 18384469 TI - Co-publishing Cochrane Library systematic reviews: journal editor and Cochrane Skin Group author experiences. PMID- 18384470 TI - Are filaggrin mutations associated with hand eczema or contact allergy?--we do not know. PMID- 18384471 TI - Q fever-related otological manifestations following chronic exposure to products of conception? PMID- 18384472 TI - Palliative therapy of giant basal cell carcinoma with the monoclonal anti epidermal growth factor receptor antibody cetuximab. PMID- 18384473 TI - The revolutionary consequences of skin disease. PMID- 18384474 TI - Antihistamine combination treatment for solar urticaria. PMID- 18384475 TI - Case-control study of risk factors associated with hepatitis C virus infection among pregnant women in hospitals of Karachi-Pakistan. AB - OBJECTIVE: To identify risk factors for hepatitis C virus (HCV) infection among pregnant women seeking antenatal care in tertiary care hospitals of Karachi, Pakistan. METHODS: We enrolled 119 cases and 238 controls. Cases were enzyme linked immunosorbent assay (ELISA III) positive pregnant women for antibodies to HCV; controls were anti-HCV ELISA negative pregnant women. RESULTS: The mean age of study subjects was 26 years (SD 5) ranging from 15 to 50 years. The mean number of pregnancies for cases was 4 (SD 3) and for controls was 3 (SD 2). Among cases an average number of injections in any month was 40%, history of hospitalization was 61% and household contact with jaundice or hepatitis was 35%. In the final multivariable logistic regression model, five or more gestations (aOR = 1.99; 95% CI = 1.08-3.33), > or =1 injection (aOR = 2.33; 95% CI = 1.38 3.91) per month, hospitalization (aOR = 1.78; 95% CI = 1.01-2.99) and household contact with jaundice/hepatitis (aOR = 3.32; 95% CI = 1.89-5.83) were independently associated with HCV. CONCLUSION: Iatrogenic exposure (health care injections, hospitalizations and gestations) is the major risk factor for transmission of HCV among pregnant women. PMID- 18384476 TI - Insecticide-treated net ownership and usage in Niger after a nationwide integrated campaign. AB - OBJECTIVES: In December 2005 and March 2006, Niger conducted nationwide integrated campaigns to distribute polio vaccine and long lasting insecticide treated nets (LLINs) to children <5 years of age. We evaluated the campaign effectiveness, net retention, insecticide-treated net (ITN) ownership, and usage. METHODS: Two nationwide cross-sectional surveys in January 2006 (dry season) and September 2006 (rainy season), using a stratified two-stage cluster sampling design. We mapped selected communities, selected households by simple random sampling, and administered questionnaires by interviewers using personal digital assistants. RESULTS: The first survey showed that ITN ownership in all households was 6.3% prior to the campaign, increasing to 65.1% after the campaign in the second survey. The second survey also showed that 73.4% of households with children <5 received an LLIN and that 97.7% of households that received > or = one LLIN retained it. The wealth equity ratio for ITN ownership in households with children <5 increased from 0.17 prior to the campaign to 0.79 afterward. During the dry season, 15.4% of all children <5 and 11.3% of pregnant women slept under an ITN, while during rainy season, 55.5% of children <5 and 48.2% of pregnant women slept under an ITN. CONCLUSIONS: Free distribution during the integrated campaign rapidly increased ITN ownership and decreased inequities between those in the highest and lowest wealth quintiles. Retention of ITNs was very high, and usage was high during malaria transmission season. However, ITN ownership and usage by vulnerable groups continues to fall short of RBM targets, and additional strategies are needed to increase ownership and usage. PMID- 18384477 TI - Helicobacter pylori isolates recovered from gastric biopsies of patients with gastro-duodenal pathologies in Cameroon: current status of antibiogram. AB - OBJECTIVE: To determine the prevalence of Helicobacter pylori in patients with gastro-duodenal pathologies and the susceptibility patterns of isolates to the currently recommended antibiotic treatment regimen used in Cameroon. METHODS: Consecutive dyspeptic patients referred to Douala General Hospital, Cameroon for endoscopy were recruited in the study. Gastric biopsies were collected from the patients and H. pylori isolated and identified following standard microbiology and biochemical techniques. Antibiotic susceptibility was determined by disk diffusion and agar dilution methods against clarithromycin, tetracycline, amoxicillin and metronidazole. Data were analysed using chi-square test and significance considered at P < 0.05. RESULTS: Seventy-one (92.2%) of the 77 patients (mean age 44.5 +/- 15.7, range 15-77 years) were positive for H. pylori. The antibiotic susceptibility rates were 56% for tetracycline, 55.3% for clarithromycin, 14.4% for amoxicillin and 6.8% for metronidazole. The prevalence of clarithromycin resistance in males vs. females was 42.1%vs. 46.7%, while for metronidazole it was 89.5%vs. 94.7% (P > 0.05). Antimicrobial susceptibility results also revealed 12 antibiotypes based on resistance to the antimicrobial agents investigated. The resistance pattern, amoxicillin and metronidazole (AMR(R) MET(R)) was the most common (23.7%) amongst the isolates. More than 60% of the isolates exhibited multi-drug resistance to three or four antibiotics. CONCLUSION: Multi-drug resistance is common against the current treatment regimen in Cameroon and, therefore, calls for urgent studies involving newer and broad spectrum antibiotics to address the problem. PMID- 18384478 TI - ST2859 serogroup A meningococcal meningitis outbreak in Nouna Health District, Burkina Faso: a prospective study. AB - We analysed cerebrospinal fluid samples from suspected meningitis cases in Nouna Health District, Burkina Faso, during the meningitis seasons of 2004-2006. Serogroup A ST2859 meningococci belonging to the ST5 clonal complex of subgroup III meningococci were the predominant causative agent. ST2859 bacteria were associated with focal outbreaks in the north of the district. While >10% of the population of an outbreak village carried ST2859, the population in the south of the district was predominantly colonised by serogroup Y ST4375 meningococci, which were associated with only sporadic cases of meningitis. Colonisation with the less virulent Y meningococci may interfere with the spread of the ST2859 to the south of the district, but there are concerns that this serogroup A clone may cause a third wave of subgroup III meningococcal disease in the African Meningitis Belt. PMID- 18384479 TI - Paediatric HIV and neurodevelopment in sub-Saharan Africa: a systematic review. AB - OBJECTIVE: To determine the degree of motor, cognitive, language and social emotional impairment related to HIV infection in children living in sub-Saharan Africa (SSA). METHODS: Literature searches using MEDLINE and PsycINFO. Additionally, the reference lists of previous reviews were checked to ensure that all eligible studies were identified. Cohen's d, a measure of effect size, was computed to estimate the level of impairment. RESULTS: Six reports met the inclusion criteria. In infancy a consistent delay in motor development was observed with a median value of Cohen's d = 0.97 at 18 months, indicating a severe degree of impairment. Mental development showed a moderate delay at 18 months, with a median value d = 0.67. Language delay did not appear until 24 months of age, d = 0.91. Less clear findings occurred in older subjects. CONCLUSION: Although HIV has been shown to affect all domains of child functioning, motor development is the most apparent in terms of severity, early onset, and persistence across age groups. However, motor development has been the most widely assessed domain while language development has been less vigorously evaluated in SSA, hence an accurate quantitative estimate of the effect cannot yet be made. PMID- 18384480 TI - Anaemia in a rural Ugandan HIV cohort: prevalence at enrolment, incidence, diagnosis and associated factors. AB - OBJECTIVES: To determine the prevalence and incidence of anaemia in HIV-positive and negative individuals; to identify risk factors for anaemia, prior to the introduction of HAART; and to determine the validity of the clinical diagnosis of anaemia. METHODS: Between 1990 and 2003, we followed a rural population based cohort of HIV-infected and uninfected participants. Prevalence and incidence of anaemia were determined clinically and by laboratory measurements. The sensitivity, specificity and predictive values of clinical diagnosis were calculated. RESULTS: The prevalence of anaemia at enrolment was 18.9% among HIV positive and 12.9% among HIV-negative participants (P = 0.065). Incidence of anaemia increased with HIV disease progression, from 103 per 1000 person-years of observation among those with CD4 counts >500 to 289 per 1000 person-years of observation among those with CD4 counts <200. Compared to laboratory diagnosis, the clinical diagnosis of anaemia had a sensitivity of 17.8%, specificity of 96.8%, a positive predictive value of 50.6% and a negative predictive value of 86.4%. Being female, low CD4 cell counts, HIV-positive, wasting syndrome, WHO stage 3 or 4, malaria, fever, pneumonia and oral candidiasis were associated with prevalent anaemia. CONCLUSIONS: Anaemia prevalence and incidence were higher among HIV-positive than negative participants. Compared to laboratory diagnosis, clinical detection of anaemia had a low sensitivity. Clinicians working in settings with limited laboratory support must be conscious of the risk of anaemia when managing HIV/AIDS patients, particularly when using antiretroviral drugs which by themselves may cause anaemia as a side effect. We recommend that haemoglobin should be measured before starting ART and monthly for the first three months. PMID- 18384481 TI - Very young children with uncomplicated falciparum malaria have higher risk of hypoglycaemia: a study from Suriname. AB - OBJECTIVE: To measure glucose kinetics and the influence of age, nutritional status and fasting duration in children with uncomplicated falciparum malaria (UFM) under the age of 5 years. METHODS: Plasma glucose concentration, endogenous glucose production (EGP) and gluconeogenesis (GNG) were measured using [6,6 (2)H(2)]glucose and (2)H(2)O in 17 very young (<3 years) and 7 older (3-5 years) Surinamese children with UFM admitted to the Distrikt Hospital Stoelmanseiland and Diakonessen Hospital Paramaribo over 17 months. RESULTS: Plasma glucose concentration was lower in the group of very young children than in the older children (P = 0.028). There were no differences in EGP and GNG between the groups. Overall GNG contributed 56% (median, range 17-87%) to EGP, with no differences between the groups (P = 0.240). Glucose clearance was lower in the older children (P = 0.026). Glucose concentration did not differ between children with weight for length/height less than -1.3 SD and children with weight for length/height greater than -1.3 SD (P = 0.266). Plasma glucose concentration was not predicted by fasting duration (P = 0.762). CONCLUSIONS: Our data suggest a higher risk of hypoglycaemia in very young children with uncomplicated malaria as plasma glucose concentration was lower in this study group. Since this could not be attributed to an impaired EGP, and because glucose clearance was lower in the older children, we presume that older children were better capable of reducing glucose utilization during fasting. Studies on glucose kinetics are feasible in very young children with malaria and give more insight in the pathophysiology of hypoglycaemia. PMID- 18384482 TI - Human and porcine neurocysticercosis: differences in the distribution and developmental stages of cysticerci. AB - OBJECTIVE: To describe and compare the clinical impacts of neurocysticercosis (NC) caused by Taenia solium in humans and pigs. METHODS: Comparative study of the brains of 16 asymptomatic pigs and 35 human NC cases (15 asymptomatic and 20 symptomatic). RESULTS: In humans, cysticerci were more frequently located in the ventricles and subarachnoid space at the base of the brain (11.8%vs. 1.6%; P = 0.001 and 25.9%vs. 0%; P < 0.0001, respectively) while in pigs, cysticerci were more frequently found in the parenchyma (44.4%vs. 7.6%; P < 0.0001). In human brains, 75.9% of the cysticerci were calcified, while in pigs all cysticerci were in the vesicular stage. CONCLUSION: The duration of infection and the host parasite relationship (such as immune reactivity and brain haemodynamics) differ between humans and pigs. This may account for the different distribution and stage of the cysticerci among humans and pigs. PMID- 18384483 TI - Toxoplasmosis with hemophagocytic syndrome after bone marrow transplantation: diagnosis at autopsy. AB - Toxoplasmosis is a rare but well recognized opportunistic infection that can occur after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Besides encephalitis, other common presentations of Toxoplasma gondii infection are interstitial pneumonitis and myocarditis. Because of its non-specific clinical and biological signs and its lethal outcome, toxoplasmosis is often misdiagnosed and only revealed at autopsy. We report a case of a postmortem diagnosis of disseminated toxoplasmosis associated with hemophagocytic syndrome, which underlines the value of necropsy in cases of death after transplantation. We also discuss clinical presentations and risk factors that lead to toxoplasmosis in allo-HSCT recipients. PMID- 18384484 TI - Solitary fibrous tumor of the breast and mammary myofibroblastoma: the same lesion? AB - Benign stromal tumors of the breast are rare mesenchymal neoplasms that have significant clinical and morphologic overlap. We report the case of a spindle cell tumor occurring in the mammary gland with mixed features of solitary fibrous tumor and mammary myofibroblastoma. The significance of this finding as well as the pathologic and radiologic diagnostic strategies used to differentiate these two lesions are presented based on a review of pertinent literature. PMID- 18384485 TI - Identification of 11 novel HLA alleles found during typing of unrelated registry donors in China. AB - Eleven novel human leukocyte antigen (HLA) alleles were identified during routine sequence-specific oligonucleotide probe (SSOP) typing (LABType; One Lambda Inc., Los Angeles, CA) of volunteers for a hematopoietic stem cell registry in Beijing, China. The new alleles were detected when one or more probes gave an unexpected reactivity pattern. PMID- 18384486 TI - A novel HLA-B*40 allele, HLA-B*4083, identified in a Korean individual. AB - HLA-B*4083 differs from the closest aligned sequence HLA-B*400601 by three nucleotide substitutions at codons 31 (ACG -->ACC), 32 (CTG -->CAG), and 41 (ACG -> GCG), resulting in two amino acid changes at residues 32 (Leu to Gln) and 41 (Thr to Ala). PMID- 18384487 TI - Mutation screening of the CARD15 gene in sarcoidosis. AB - CARD15 was first identified as a susceptibility gene for Crohn's disease. More recently, CARD15 mutations were shown to be associated with the pediatric granulomatous inflammatory diseases, Blau syndrome and early-onset sarcoidosis (EOS). The aim of the present study was to evaluate whether CARD15 variants also play a role in patients with ordinary sarcoidosis other than EOS. We enrolled 135 Japanese sarcoidosis patients with uveitis as well as 95 healthy individuals and performed mutation analysis by direct sequencing of CARD15 exon 4. Direct DNA sequencing in the sarcoidosis patients showed eight CARD15 variants, including five novel mutations (13402C>T, 13543C>T, 13775C>A, 13937G>A, and 14079C>T). Compared with healthy individuals, CARD15 mutations are not common in the Japanese patients with sarcoidosis. Based on the results, we examined the clinical manifestations in patients with sarcoidosis according to their CARD15 mutations. Sarcoidosis patients with these mutations have no specific clinical features with regard to course of the disease or disease severity. Our results indicate that in general, CARD15 mutations may not contribute to the risk of sarcoidosis. PMID- 18384488 TI - Lymphotoxin alpha LTA+496C allele is a risk factor for periodontitis in patients with coronary artery disease. AB - Periodontitis and coronary artery disease (CAD) are inflammatory diseases and associated with each other. The major histocompatibility complex (MHC) region carries genes involved in immune response and inflammation. We investigated whether the MHC genes correlate with the presence of periodontitis or with the occurrence of periodontal pathogens in patients with CAD. Blood and saliva samples from CAD patients (n = 106) were collected at the time of hospitalization. Nine MHC genetic markers [human leukocyte antigen (HLA)-A, HLA B, HLA-DRB1, lymphotoxin alpha (LTA) +253(a/g), +496(C/T), +633(c/g), +724(C/A), C4A and C4B)] were typed. Based on panoramic tomography, patients were categorized into nonperiodontitis and periodontitis groups. Two major periodontal pathogens, Aggregatibacter (Actinobacillus) actinomycetemcomitans and Porphyromonas gingivalis, were cultivated and polymerase chain reaction-amplified from salivary samples. Serum immunoglobulin (Ig)A and IgG antibody levels to these pathogens were measured. In the univariate analysis, LTA+496C allele (OR = 5.29; 95% CI = 2.07-13.51, P = 0.00027), and the occurrence of P. gingivalis in saliva (OR = 4.74; 95% CI = 1.64-13.70; P = 0.002) were more frequent in periodontitis when compared with nonperiodontitis. Similarly, serum IgA antibody level against the pathogen was increased in periodontitis (P = 0.048). In the multiple logistic regression analysis, when a wide range of covariates was included, the LTA+496C allele (OR = 10.87; 95% CI = 3.23-36.60; P = 0.00012) and the elevated serum IgA antibody level against P. gingivalis (OR = 1.56; 95% CI = 1.05-2.30; P = 0.026) remained as significant risk factors for periodontitis. In conclusion, the major finding of this study is that the LTA+496C allele is associated with periodontitis in patients with CAD. PMID- 18384489 TI - Stability training of the lumbo-pelvo-hip complex influence stiffness of the hamstrings: a preliminary study. AB - An analysis of data obtained in an experiment investigating the influence of stability training of the lumbo-pelvo-hip complex (LPHC) on stiffness of the hamstrings is presented. Randomized controlled trial. The study included 30 subjects (aged 18-42 years) with increased stiffness of the hamstrings at baseline. Over a period of 4 weeks, stability training aiming to activate the deep stabilizing muscle subsystem and to integrate its action with the superficial subsystem was introduced in the experimental group. The control group remained unaffected. Three series of measurements were applied (baseline, after 2 weeks, and after 4 weeks). A digital inclinometer was used to measure outcomes of passive knee extension in the supine test. In the experimental group, a tendency to decrease stiffness of the hamstrings was observed. It was the opposite in the control group. Significant intra-group differences in the experimental group between series 1 and 3 measurements for both the right and left lower extremities were revealed. Stability training of the LPHC showed a tendency to be effective in reducing stiffness of the hamstrings. PMID- 18384490 TI - Effect of hamstring flexibility on isometric knee flexion angle-torque relationship. AB - The purpose of this study was to examine the relationship between hamstring flexibility and knee flexion angle-torque relationship. Hamstring flexibility was assessed in 20 subjects (10 men, 10 women) using the straight leg raise (SLR) and active knee extension (AKE) tests. Isometric knee flexion strength was measured at five knee flexion angles while subjects were seated with the test thigh flexed 40 degrees and the trunk flexed 80 degrees . Lower extremities were classified as tight or normal based on the SLR and AKE tests. Peak knee flexion torque, angle of peak torque, and angle-torque relationship were compared between flexibility groups. Peak knee flexion torque was not different between tight and normal groups (SLR P=0.82; AKE P=0.68) but occurred in greater knee flexion (shorter muscle length) in the tight group compared with the normal group (SLR P<0.01; AKE P<0.05). The tight group had higher torque than the normal group at the shortest muscle length tested but lower torque at longer muscle lengths (SLR P<0.001; AKE P<0.001). In conclusion, the angle-torque relationship was shifted to the left in less flexible hamstrings such that knee flexion torque was increased at short muscle lengths and decreased at long muscle lengths when compared with more flexible hamstrings. PMID- 18384491 TI - Personal and contextual determinants of elite young athletes' persistence or dropping out over time. AB - Two studies were conducted among elite young judokas to examine (a) whether those who persisted in national training centers (n=52) were different from dropouts (n=52) in their perceptions of coach-, parent-, and peer-induced motivational climates, goal orientations, self-perceptions, perceived competence, and intention of dropping out, and (b) whether these variables varied during the persisting athletes' (n=82) first 2 years in these centers. Compared with persisting athletes, dropouts perceived the roles of coaches, parents, and peers as less task-involving, were less task-oriented, and intended more to drop. The association of peer-, coach-, and parent-induced task-involving climates predicted athletes' persistence. During the 2 years, persisting athletes' perceptions of coach-, parent-, and peer-induced task-involving climates decreased, while perceptions of a coach-induced ego-involving climate and the intention of dropping out increased in spite of more positive self-perceptions. Gender differences favoring males were observed for self-perceptions only. These results stress the importance for all the agents of the athletes' social environment to promote task-involving climates, because such climates appear to be naturally prone to degradation in the context of elite competition. The results also shed light on some high-level athletes' characteristics regarding motivational dispositions and self-concept. PMID- 18384492 TI - Skeletal muscle fiber-type distribution and habitual physical activity in daily life. AB - The capacity to perform physical activity largely depends on physical fitness. Muscle fiber-type distribution (Muscle(FTD)) is associated with physical fitness and may influence the capacity to perform physical activity. The purpose of this study was to determine whether habitual physical activity in daily life (PA(DL)) and Muscle(FTD) are related. Thirty-eight healthy non-athletes (31 women, 7 men) were recruited. PA(DL) was measured twice for 14 days using a tri-axial accelerometer for movement registration (Tracmor). From Tracmor output, the proportion of time subjects were physically active at low, moderate, and high intensities was determined (%Low, %Moderate, and %High, respectively). A total activity index (PA(index)) and sub-scores on work, leisure-time and sports were obtained using the Baecke questionnaire. Muscle(FTD) was determined using immuno fluorescence against respective myosin heavy chain isoforms. No relationship was observed between PA(DL) and Muscle(FTD). %Low, %Moderate, and %High, as well as PA(index) and its sub-scores, were not related to Muscle(FTD) either. The time spent on sports was associated with the proportion of type I and II(X) fibers (P=0.06 and P<0.01, respectively). In conclusion, Muscle(FTD) probably cannot explain why some people are more prone to engaging in physical activities than others. PMID- 18384493 TI - Muscular strength, functional performances and injury risk in professional and junior elite soccer players. AB - Muscle strength and anaerobic power of the lower extremities are neuromuscular variables that influence performance in many sports activities, including soccer. Despite frequent contradictions in the literature, it may be assumed that muscle strength and balance play a key role in targeted acute muscle injuries. The purpose of the present study was to provide and compare pre-season muscular strength and power profiles in professional and junior elite soccer players throughout the developmental years of 15-21. One original aspect of our study was that isokinetic data were considered alongside the past history of injury in these players. Fifty-seven elite and junior elite male soccer players were assigned to three groups: PRO, n=19; U-21, n=20 and U-17, n=18. Players benefited from knee flexor and extensor isokinetic testing consisting of concentric and eccentric exercises. A context of lingering muscle disorder was defined using statistically selected cut-offs. Functional performance was evaluated throughout a squat jump and 10 m sprint. The PRO group ran faster and jumped higher than the U-17 group (P<0.05). No significant difference in isokinetic muscle strength performance was observed between the three groups when considering normalized body mass parameters. Individual isokinetic profiles enabled the identification of 32/57 (56%) subjects presenting lower limb muscular imbalance. Thirty-six out of 57 players were identified as having sustained a previous major lower limb injury. Of these 36 players, 23 still showed significant muscular imbalance (64%). New trends in rational training could focus more on the risk of imbalance and implement antagonist strengthening aimed at injury prevention. Such an intervention would benefit not only athletes recovering from injury, but also uninjured players. An interdisciplinary approach involving trainers, a physical coach, and medical staff would be of interest to consider in implementing a prevention programme. PMID- 18384494 TI - Environmental characteristics relevant to young people's use of sports facilities: a review. AB - Several countries spend large sums of public money building sports facilities. Still, our knowledge about the effect of these means is limited. The objectives in this review are (1) to identify relevant knowledge of how sports facilities influence young people's physical activity, (2) to clarify the status of examined and non-examined factors and (3) to propose a conceptual model to guide future judgments. Relevant studies were located from searches in computerized databases and the bibliography of reviews. Ecological system theory uses the theoretical framework. The findings encompass 239 independent samples located in 43 studies, primarily from the field of public health. Based on these and the framework, we have developed a modified, conceptual model. The model stresses the interaction between the demographic, personal and interpersonal characteristics of the user, the consideration of access and facility characteristics and the level where this takes place. The most consistent data were found at the mesolevel and were correlated to access and user characteristics. Knowledge gaps were found at the microlevel and with regard to facility characteristics. We also know too little with respect to subgroups' different behavior in various physical settings. Finally, there is a need to confirm/disconfirm less consistent findings. PMID- 18384495 TI - Judgment of other people's facial expressions of emotions is influenced by their concurrent affective hand movements. AB - We investigated whether emotional information from facial expression and hand movement quality was integrated when identifying the expression of a compound stimulus showing a static facial expression combined with emotionally expressive dynamic manual actions. The emotions (happiness, neutrality, and anger) expressed by the face and hands were either congruent or incongruent. In Experiment 1, the participants judged whether the stimulus person was happy, neutral, or angry. Judgments were mainly based on the facial expressions, but were affected by manual expressions to some extent. In Experiment 2, the participants were instructed to base their judgment on the facial expression only. An effect of hand movement expressive quality was observed for happy facial expressions. The results conform with the proposal that perception of facial expressions of emotions can be affected by the expressive qualities of hand movements. PMID- 18384496 TI - Decision-making styles and physiological correlates of negative stress: is there a relation? AB - There are 5 independent, although not mutually exclusive, styles used by military officers when making decisions: rational, intuitive, dependent, avoidant, and spontaneous (Scott & Bruce, 1995). The rational and intuitive styles are part of the cognitive style concept, whereas the conceptual foundation of the remaining 3 styles is less clear. The dependent and the avoidant styles were expected to be accompanied by increased levels of negative stress, while this was not expected of the remaining 3 styles. This hypothesis was put to test. Twenty-three male Swedish Army majors were asked to make decisions in 2 different military situations. Their decision styles were measured by the General Decision Making Style inventory (GDMS; Scott & Bruce, 1995). Saliva cortisol release indicated negative stress and was measured on 4 occasions: twice before, after the first decision, and after the second decision. The avoidant style was related to negative stress, as predicted. There was, however, no difference between the base line measurements and the after-decision measurements, suggesting that officers with an avoidant decision style may, in general, have higher levels of cortisol secretion (i.e., being more of the worrying kind). PMID- 18384497 TI - Evaluation of a Swedish version of the Job Stress Survey. AB - The present study assesses and evaluates the psychometric properties of a Swedish version of the Job Stress Survey (JSS; Spielberger, 1991; Spielberger & Vagg, 1999). This instrument is constructed to measure generic sources of occupational stress encountered by employees in a wide variety of work settings, settings that often result in psychological strain. The JSS was administered to metal assembly industry workers and medical service personnel in northern Sweden (n= 1186). The exploratory factor analysis showed that there is a high similarity between the present Swedish version and the original American version. Internal reliabilities of the scales, as well as test-retest reliabilities were shown to be high, and concurrent validity, as examined by comparisons with the Perceived Stress Questionnaire Index (Levenstein et al., 1993) was found to be satisfactory. The consistency of these findings is discussed with particular focus on groups of employees, gender, and cross-cultural evaluations. PMID- 18384498 TI - Reduced fertility in high-yielding dairy cows: are the oocyte and embryo in danger? Part II. Mechanisms linking nutrition and reduced oocyte and embryo quality in high-yielding dairy cows. AB - Dairy cow fertility has been declining during since the mid-80s and this has given rise to numerous scientific studies in which important parts of the pathogenesis are elucidated. Reduced oocyte and embryo quality are acknowledged as major factors in the widely described low conception rates and in the high prevalence of early embryonic mortality. Apart from the importance of the negative energy balance (NEB) and the associated endocrine and metabolic consequences, there is a growing attention towards the effect of the milk yield promoting diets which are rich in energy and protein. Starch-rich diets can improve the energy status and thus the ovarian activity in the early postpartum period but the oocyte and embryo quality can suffer from such insulinogenic diets. Supplementation of dietary fat has a similar dual effect with a beneficial stimulation of the ovarian steroid production while the oocyte and the embryo display an altered energy metabolism and excessive lipid accumulation. High protein diets can elevate the ammonia and urea concentrations in the blood, leading to changed intrafollicular, oviductal and uterine environments. Oocytes and embryos are highly sensitive to such changes in their microenvironment, possibly leading to a disturbed maturation, fertilization or early cleavage. Several nutrition-linked mechanisms, through which oocyte and/or embryo quality can be affected in modern dairy cows, well after the period of NEB, are proposed and comprehensively reviewed in the present report. PMID- 18384499 TI - Reduced fertility in high-yielding dairy cows: are the oocyte and embryo in danger? Part I. The importance of negative energy balance and altered corpus luteum function to the reduction of oocyte and embryo quality in high-yielding dairy cows. AB - Fertility in high yielding dairy cows is declining, and there is increasing evidence to presume that oocyte and embryo quality are major factors in the complex pathogenesis of reproductive failure. In this report we present an overview of possible mechanisms linking negative energy balance (NEB) and deficiencies in oocyte and embryo developmental competence; specifically, in the high producing dairy cow. Changes in follicular growth patterns during a period of NEB can indirectly affect oocyte quality. The endocrine and biochemical changes, which are associated with NEB, are reflected in the microenvironment of the growing and maturing female gamete, and likely result in the ovulation of a developmentally incompetent oocyte. Even after an oocyte is successfully ovulated and fertilized, a full-term pregnancy is still not guaranteed. Inadequate corpus luteum function, associated with reduced progesterone, and probably also low insulin-like growth factor concentrations, can cause a suboptimal microenvironment in the uterus that is incapable of sustaining early embryonic life. This may partly account for the low conception rates and the high incidence of early embryonic mortality in high yielding dairy cows. PMID- 18384500 TI - Housing during early pregnancy affects fertility and behaviour of sows. AB - From 1.1.2013 EU legislation will permit stall-housing of dry sows for only 4 weeks post-service. Stalls are associated with compromised animal welfare, but group housing comes with risks from social confrontations. Studies on housing effects on fertility have produced contradictory results as systems are inconsistently defined. Effects of housing on pregnancy rate 28 days post-service (P28), early disruption of pregnancy (EDP) and behaviour were investigated in 12 replicates of 40 sows. Half of the animals were stalled (treatment S, mean parity 2.4) and half group-housed adjacent on 5.1 m(2) deep litter per sow (treatment G, mean parity 2.3). Pregnancy was detected using real-time ultrasound. An EDP diagnosis required a drop of salivary progesterone concentration from >15.9 to <15.9 mmol/l in a series of samples taken on days 17, 20, 24 and 27 post-service. Behaviour was scan sampled on day 27 for 120 min with 10-min intervals. Data was analysed using mixed regression models in MLwiN. Treatment S increased the odds for P28 with 2.3 and the pregnancy rate of a treatment group with 0.1 compared with G. G increased the odds for EDP with 4.6. Treatment did not affect total exploratory or total passive behaviour. S sows explored floor and fixtures (not bedding) more and performed more passive sitting than G sows. Social stress may have caused the impaired reproductive performance in G sows. S sows showed behavioural signs of decreased welfare, but no corresponding reproductive effects. An interdisciplinary approach is needed when evaluating the appropriateness of housing systems. PMID- 18384501 TI - Patient-controlled analgesia in chronic pain patients: experience with a new device designed to be used with implanted programmable pumps. AB - BACKGROUND: Intrathecal drug delivery using implantable pumps is an effective method to control stable chronic pain. However, the appropriate alleviation of unpredictable pain fluctuations remains challenging. A possible solution is the use of patient-controlled analgesia (PCA) by means of a specific device--the personal therapy manager (PTM)--designed to be used with implanted programmable pumps. METHODS: A multicenter (n = 17), open-label registry recording 168 patients suffering from chronic pain with unsatisfactory control of pain episodes was initiated. While 79 patients (47%) (group A) already carried an implanted pump at enrollment, all other patients were implanted after being registered in the study (group B). Parameters assessed included pain relief (visual analog score, VAS), quality of life (EQ-5D), patient and physician satisfaction, medication use, PTM programming parameters and adverse events. Final follow-up was at 12 months. RESULTS: At 12 months, there was a significant reduction (29%) of the overall average VAS compared with baseline (P < 0.01) in patients with newly implanted devices (group B). All patients tended to decrease the concomitant pain medication and the quality of life tended to improve (10% on the EQ-5D scale). In total, 85% of patients were satisfied with the PTM. No serious adverse events related to the use of the PTM device were observed. CONCLUSIONS: Patient-controlled analgesia using a PTM with a programmable, implantable pump system is an effective therapy for the treatment of chronic pain and allows patients to feel that they have more control over unpredictable pain fluctuations. PMID- 18384502 TI - AtCXXS: atypical members of the Arabidopsis thaliana thioredoxin h family with a remarkably high disulfide isomerase activity. AB - The Arabidopsis thaliana thioredoxin subgroup h III is composed of four members and includes the two monocysteinic (CXXS) thioredoxins encoded by the genome. We show that AtCXXS1 is the ortholog of monocysteinic thioredoxins present in all higher plants. In contrast, unicellular algae and the moss Physcomitrella patens do not encode monocysteinic thioredoxin. AtCXXS2, the second monocysteinic thioredoxin of Arabidopsis has no ortholog in any other higher plants. It probably appeared recently by duplications of a dicysteinic thioredoxin of the same subgroup h III. Both monocysteinic thioredoxins show a low disulfide reductase activity in vitro but are very efficient as disulfide isomerases in RNAse refolding tests. The possible interactions of these proteins with the glutathione glutaredoxin pathway are discussed on the basis of recent papers. PMID- 18384503 TI - Nitrogen stress and the expression of asparagine synthetase in roots and nodules of soybean (Glycine max). AB - The difficulty of assaying asparagine synthetase (AS) (EC 6.3.5.4) activity in roots of soybean has been circumvented by measuring expression of the AS genes. Expression of three soybean asparagine synthetase (SAS) genes (SAS1, SAS2 and SAS3) was observed in roots of non-nodulated soybean plants cultivated on nitrate. Expression of these genes was reduced to very low levels within days after submitting the plants to a N-free medium. The subsequent return to a complete medium (containing nitrate) restored expression of all three AS genes. Roots of nodulated plants, where symbiotic nitrogen fixation was the exclusive source of N (no nitrate present), showed very weak expression of all three AS genes, but on transfer to a nitrate-containing medium, strong expression of these genes was observed within 24 h. In nodules, all three genes were expressed in the absence of nitrate. Under conditions that impair nitrogen fixation (nodules submerged in aerated hydroponics), only SAS1 expression was reduced. However, in the presence of nitrate, an inhibitor of N(2) fixation, SAS1 expression was maintained. High and low expressions of AS genes in the roots were associated with high and low ratios of Asn/Asp transported to the shoot through xylem. It is concluded that nitrate (or one of its assimilatory products) leads to the induction of AS in roots of soybean and that this underlies the variations found in xylem sap Asn/Asp ratios. Regulation of nodule AS expression is quite different from that of the root, but nodule SAS1, at least, appears to involve a product of N assimilation rather than nitrate itself. PMID- 18384505 TI - The reactive oxygen-driven tumor: relevance to melanoma. AB - In melanoma, at least four major signaling abnormalities have been described. They include beta-catenin deregulation (mutation/mislocalization), p16 loss, MAP kinase activation, and Akt activation. In this review, we discuss the role of the fourth pathway, known as the reactive oxygen driven tumor. The role of reactive oxygen in tumorigenesis is likely to relate to virtually all forms of cancer, and lends itself to specific therapies. These include blockade of reactive oxygen, resulting in decreased activation of NF-kappaB, which should sensitize tumors to chemotherapy and radiation. The phenotype of the reactive oxygen driven tumor can be monitored using available markers already in use in most hospital laboratories. PMID- 18384506 TI - High-density seedling expression system for the production of bioactive human cardiotrophin-1, a potential therapeutic cytokine, in transgenic tobacco chloroplasts. AB - Histidine-tagged human cardiotrophin-1 (hCT-1), a recently discovered cytokine with excellent therapeutic potential, was expressed in tobacco chloroplasts under the transcriptional and translational control of two different promoters (rrn and psbA) and 5'-untranslated regions (5'-UTRs) (psbA and phage T7 gene 10). The psbA 5'-UTR promotes recombinant hCT-1 (rhCT-1) accumulation in chloroplasts at higher levels (eight-fold) than those obtained for the phage T7 gene 10 5'-UTR, regardless of the promoter used, indicating that the correct choice of translational control element is most important for protein production in chloroplasts. The maximum level of rhCT-1 achieved was 1.14 mg/g fresh weight (equivalent to 5% of total soluble protein) with the psbA promoter and 5'-UTR in young leaves harvested after 32 h of continuous light, although the bioactivity was significantly lower (approximately 35%) than that of commercial hCT-1. However, harvesting in the dark or after 12 h of light did not result in a significant decrease in the bioactivity of rhCT-1, suggesting that 32 h of over lighting affects the biological activity of rhCT-1. Because high levels of rhCT-1 accumulation took place mainly in young leaves, it is proposed that seedlings should be used in a 'closed system' unit, yielding up to 3.2 kg per year of rhCT 1. This amount would be sufficient to meet the estimated annual worldwide needs of hCT-1 for liver transplantation surgery in a cost-effective manner. Furthermore, our strategy is an environmentally friendly method for the production of plant-based biopharmaceuticals. PMID- 18384507 TI - Transgenic tomato plants alter quorum sensing in plant growth-promoting rhizobacteria. AB - Two Gram-negative, plant growth-promoting rhizobacteria (PGPRs), denominated as M12 and M14, were classified by 16S rDNA sequencing as Burkholderia graminis species. Both strains were shown to produce a variety of N-acyl-homoserine lactone (AHL) quorum sensing (QS) signalling molecules. The involvement of these molecules in plant growth promotion and the induction of protection against salt stress was examined. AHL production was evaluated in vitro by thin-layer chromatography using AHL biosensors, and the identity of the AHLs produced was determined by liquid chromatography-tandem mass spectrometry. The in situ production of AHLs by M12 and M14 in the rhizosphere of Arabidopsis thaliana plants was detected by co-inoculation with green fluorescent protein-based biosensor strains and confocal laser scanning microscopy. To determine whether plant growth promotion and protection against salt stress were mediated by QS, these PGPRs were assayed on wild-type tomato plants, as well as their corresponding transgenics expressing YenI (short-chain AHL producers) and LasI (long-chain AHL producers). In wild-type tomato plants, only M12 promoted plant growth, and this effect disappeared in both transgenic lines. In contrast, M14 did not promote growth in wild-type tomatoes, but did so in the LasI transgenic line. Resistance to salt stress was induced by M14 in wild-type tomato, but this effect disappeared in both transgenic lines. The strain M12, however, did not induce salt resistance in wild-type tomato, but did so in LasI tomato plants. These results reveal that AHL QS signalling molecules mediate the ability of both PGPR strains M12 and M14 to promote plant growth and to induce protection against salt stress. PMID- 18384508 TI - Soybean WRKY-type transcription factor genes, GmWRKY13, GmWRKY21, and GmWRKY54, confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants. AB - WRKY-type transcription factors have multiple roles in the plant defence response and developmental processes. Their roles in the abiotic stress response remain obscure. In this study, 64 GmWRKY genes from soybean were identified, and were found to be differentially expressed under abiotic stresses. Nine GmWRKY proteins were tested for their transcription activation in the yeast assay system, and five showed such ability. In a DNA-binding assay, three proteins (GmWRKY13, GmWRKY27 and GmWRKY54) with a conserved WRKYGQK sequence in their DNA-binding domain could bind to the W-box (TTGAC). However, GmWRKY6 and GmWRKY21, with an altered sequence WRKYGKK, lost the ability to bind to the W-box. The function of three stress-induced genes, GmWRKY13, GmWRKY21 and GmWRKY54, was further investigated using a transgenic approach. GmWRKY21-transgenic Arabidopsis plants were tolerant to cold stress, whereas GmWRKY54 conferred salt and drought tolerance, possibly through the regulation of DREB2A and STZ/Zat10. Transgenic plants over-expressing GmWRKY13 showed increased sensitivity to salt and mannitol stress, but decreased sensitivity to abscisic acid, when compared with wild-type plants. In addition, GmWRKY13-transgenic plants showed an increase in lateral roots. These results indicate that the three GmWRKY genes play differential roles in abiotic stress tolerance, and that GmWRKY13 may function in both lateral root development and the abiotic stress response. PMID- 18384509 TI - Differential regulation of grain sucrose accumulation and metabolism in Coffea arabica (Arabica) and Coffea canephora (Robusta) revealed through gene expression and enzyme activity analysis. AB - * Coffea arabica (Arabica) and Coffea canephora (Robusta) are the two main cultivated species used for coffee bean production. Arabica genotypes generally produce a higher coffee quality than Robusta genotypes. Understanding the genetic basis for sucrose accumulation during coffee grain maturation is an important goal because sucrose is an important coffee flavor precursor. * Nine new Coffea genes encoding sucrose metabolism enzymes have been identified: sucrose phosphate synthase (CcSPS1, CcSPS2), sucrose phosphate phosphatase (CcSP1), cytoplasmic (CaInv3) and cell wall (CcInv4) invertases and four invertase inhibitors (CcInvI1, 2, 3, 4). * Activities and mRNA abundance of the sucrose metabolism enzymes were compared at different developmental stages in Arabica and Robusta grains, characterized by different sucrose contents in mature grain. * It is concluded that Robusta accumulates less sucrose than Arabica for two reasons: Robusta has higher sucrose synthase and acid invertase activities early in grain development - the expression of CcSS1 and CcInv2 appears to be crucial at this stage and Robusta has a lower SPS activity and low CcSPS1 expression at the final stages of grain development and hence has less capacity for sucrose re-synthesis. Regulation of vacuolar invertase CcInv2 activity by invertase inhibitors CcInvI2 and/or CcInvI3 during Arabica grain development is considered. PMID- 18384510 TI - Limits to local adaptation in six populations of the annual plant Diodia teres. AB - * Local adaptation is common, but tests for adaptive differentiation frequently compare populations from strongly divergent environments, making it unlikely that any influence of stochastic processes such as drift or mutation on local adaptation will be detected. Here, the hypothesis that local adaptation is more likely to develop when the native environments of populations are more distinct than when they are similar was tested. * A reciprocal transplant experiment including two populations from each of three habitats was conducted to determine the pattern of local adaptation. In addition to testing for local adaptation at the population level, the hypothesis was tested that local adaptation is more common between populations from different habitats than between populations from the same habitat. * Local adaptation was not common, but more evidence was found of local adaptation between populations from different habitats than between populations from the same habitat. Two instances of foreign genotype fitness advantage confirm that stochastic processes such as drift can limit local adaptation. * These results are consistent with the hypothesis that stochastic processes can inhibit local adaptation but are more likely to be overwhelmed by natural selection when populations occur in divergent environments. PMID- 18384511 TI - Galectin-3: a useful biomarker for differential diagnosis of brain tumors. AB - Galectin-3 (gal-3) is a 31 kDa beta-galactoside-binding lectin that is immunohistochemically expressed in macrophages, lymphocytes, and endothelial cells, and also in some neoplastic cells. Gal-3's expression in and significance to brain tumors has not been fully addressed. Here, we investigated its immunohistochemical expression in 409 cases of surgically resected primary brain tumors, including various glioneuronal tumors, pituitary adenomas, meningiomas and Schwannomas, among others. In normal brain tissues, gal-3 was robustly expressed in normal astrocytes, endothelial cells and macrophages. It showed consistent and diffuse positivity in 100% of the pilocytic astrocytomas, pleomorphic xanthoastrocytomas (PXA), Schwannomas, meningiomas, capillary hemangioblastomas, as well as in ependymomas, but it was completely negative in the diffuse astrocytomas, anaplastic astrocytomas, both low- and high-grades of the oligodendrogliomas, central neurocytomas, and medulloblastomas. Definitely positive but heterogeneous expression was found in various tumors including subependymal giant cell astrocytomas (SEGA), classic glioblastoma multiforme, anaplastic oligoastrocytomas, CNS primitive neuroectodermal tumors (CNS PNETs), and hemangiopericytomas. Eighty percent of small cell glioblstomas were completely negative, but 20% showed heterogeneous positivity for gal-3. Focal positivity for gal-3 was also found in dysembryoplastic neuroepithelial tumors (DNTs) and gangliogliomas, in which the positive cells were the astrocytic component. On the basis of our immunohistochemical data in conjunction with previous reports, we therefore conclude that gal-3 is differentially expressed in various brain tumors, and thereby, is a helpful biomarker in making differential diagnoses, especially in cases where a morphological diagnosis is controversial. PMID- 18384512 TI - Neuronal toxicity of expanded polyglutamine depends on intracellular distribution in addition to the expression level. AB - The proteins that accumulate in pathologic lesions of neurodegenerative disorders are thought to be closely associated with neuronal cell damage. However, whether or not the formation of cytoplasmic or nuclear inclusions by expanded polyglutamine (polyQ) is directly toxic to neurons has been controversial to date. We prepared a culture model system in which polyQ tracts were transfected into Neuro2a, cells of neuronal origin, to study novel factors involved in cell toxicity of polyQ tracts to neuronal cells. Pathogenic polyQ tracts of 79 repeats (Q79C) when expressed in cytoplasm of Neuro2a cells changed in their intracellular distribution patterns from homogeneous, via punctate aggregates, to massive aggregates with incubation time. Some polyQ tracts formed nuclear inclusions. Cytoplasmic massive aggregates of Q79C tended to be associated with apoptotic fate of Neuro2a cells. Cells exhibiting cytoplasmic massive inclusions had the highest expression level of polyQ tracts among cells with four patterns of intracellular distribution. The elevation in the expression levels of polyQ tracts was not due to the difference in the initial transfection efficiency. When compared among cells expressing polyQ tracts at similar levels, damages were most remarkable in cells with cytoplasmic massive aggregate in terms of shrunken cellular and nuclear sizes. Cells with the other patterns of polyQ tract distribution such as cytoplasmic homogeneous, cytoplasmic punctate and nuclear inclusions were relatively spared. These data suggest that the severity of cell damages depends on the type of intracellular distribution of polyQ tracts, in addition to the expression level of polyQ tracts. PMID- 18384513 TI - Characterization of Numb expression in astrocytomas. AB - During early neurodevelopment, asymmetric segregation of Numb in mitotic progenitor cells influences the fate of daughter cells, whereby one daughter retains the progenitor phenotype while the other proceeds along a differentiation pathway. Numb has also been reported to function as a tumor suppressor in breast cancers and medulloblastomas. Given its role in maintaining neural progenitor pools in animal models and its reported role as a tumor suppressor, Numb could potentially contribute to astrocytoma oncogenesis. We characterized Numb expression in both human astrocytoma tissue samples and glioblastoma cell lines. We found that Numb is expressed in all grades of astrocytomas, being predominantly cytoplasmic in higher-grade astrocytomas but nuclear in pilocytic astrocytomas. Numb is also present in normal neurons, but not in normal astrocytes. In cultured glioblastoma cells, Numb concentrates in the perinuclear region and process tips. Numb expression in astrocytomas recapitulates that of progenitor cells during neurodevelopment, and suggests a role for Numb in astrocytoma oncogenesis. PMID- 18384514 TI - Radiation-induced glioblastoma in a medulloblastoma patient: a case report with molecular features. AB - We report a case of glioblastoma (GBM) occurring 8 years after radiation therapy for a medulloblastoma. A 15-year-old boy underwent surgery and radiotherapy for a medulloblastoma and 8 years later he developed a second tumor at the same site. The second lesion showed different histological and molecular features, was diagnosed as a glioblastoma and fulfilled the criteria of radiation-induced neoplasm. Mutational analysis of the p53 gene showed a C to G transition at codon 176 in tumor DNA. LOH was detected at 17p and 19q. The tumor also showed O6 methylguanine-DNA methyl-transferase (MGMT) promoter methylation and no amplification of EGF receptor. In conclusion, the radiation-induced MGMT hyper methylation and p53 mutations may have a role in the development of a subgroup of radio-induced glioma (RIG), suggesting that these molecular alterations directly cooperate in the genesis of the post-irradiation GBM. Moreover RIGs seem to be a heterogeneous group of tumors that may resemble either primary or secondary GBM. PMID- 18384515 TI - Portal-systemic shunt encephalopathy presenting with diffuse cerebral white matter lesion: an autopsy case. AB - We report herein an autopsy case of portal-systemic encephalopathy (PSE) presenting with diffuse tissue rarefaction in the cerebral deep white matter. Clinically, the patient showed recurrent episodes of unconsciousness, abnormal behavior and urinary incontinence, as well as flapping tremor. Cognitive impairment and peripheral neuropathy developed following recurrent episodes. Although conventional arterial portography revealed a small portal-systemic collateral vessel of a left gastro-renal venous shunt, abdominal CT and liver biopsy showed no evidence of liver cirrhosis and serum ammonia level showed a mild increase. T2-weighted MRI demonstrated symmetrical signal hyperintensities in the deep white matter. Neuropathological findings showed Alzheimer type II astrocytes in the deep layers of the cerebral cortices and severe tissue rarefaction with no or slight reactive astrocytosis in the subcortical and deep white matter. These white matter changes have been reported infrequently in patients with PSE. The present case suggests that chronic PSE without liver cirrhosis may develop diffuse white matter lesions. PMID- 18384516 TI - Case of fulminant-SSPE associated with measles genotype D7 from India: an autopsy study. AB - Subacute sclerosing panencephalitis (SSPE), a post-measles progressive neurological disorder is still common in India because of indifferent vaccination compliance. However, the acute fulminant form of SSPE is extremely rare. An unusual case of fulminant SSPE in an 18-year-old man from south India with an ultra-short course of 19 days presenting with hemiparesis in absence of myoclonus and progressive cognitive decline, is reported. MRI showed frontal and parieto occipital demyelination extending to nuclear areas. Antimeasles antibodies were demonstrable in CSF and serum with oligoclonal bands in CSF despite normal CSF protein and cell count. At autopsy, unlike classical SSPE, oligodendroglia containing measles viral antigen was sparse despite florid necrotizing leukoencephalitis with acute demyelination. Measles virus was isolated from the brain with hypermutation in M gene confirming the diagnosis. Phylogenetic analysis of the viral genotype indicated that it belonged to D7 genotype which is considered rare in India. PMID- 18384517 TI - A malonyl-CoA-dependent switch in the bacterial response to a dysfunction of lipid metabolism. AB - Bacteria stringently regulate the synthesis of their membrane phospholipids, but the responsible regulatory mechanisms are incompletely understood. Bacillus subtilis FabF, the target of the mycotoxin cerulenin, catalyses the condensation of malonyl-ACP with acyl-ACP to extend the growing acyl chain by two carbons. Here we show that B. subtilis strains containing the fabF1 allele, which codes for the cerulenin-insensitive protein FabF[I108F], overexpressed several genes involved in fatty acid and phospholipid biosynthesis (the fap regulon) and had significantly elevated levels of malonyl-CoA. These results pinpointed FabF[I108F] as responsible for the increased malonyl-CoA production, which in turn acts as an inducer of the fap regulon by impairing the binding of the FapR repressor to its DNA targets. Synthesis of acyl-ACPs by a cell-free fatty acid system prepared from fabF1 cells showed the accumulation of short- and medium chain acyl-ACPs. These results indicate that the acyl-ACP chain length acceptance of FabF[I108F] is biased towards shorter acyl-ACPs. We also provide evidence that upregulation of FabF[I108F] is essential for survival and for resistance to cerulenin of fabF1 cells. These findings indicate that malonyl-CoA is a key molecule to monitor lipid metabolism functioning and trigger appropriate genetic and biochemical adjustments to relieve dysfunctions of this essential metabolic pathway. PMID- 18384518 TI - The Aspergillus nidulans FcyB cytosine-purine scavenger is highly expressed during germination and in reproductive compartments and is downregulated by endocytosis. AB - We cloned and characterized an Aspergillus nidulans gene, called fcyB, encoding the closest homologue to the yeast Fcy2p/Fcy21p permeases. Deletion of fcyB (DeltafcyB) does not affect growth, development, reproduction or bulk purine uptake, but eliminates the leaky growth on purines of DeltaazgADeltauapCDeltauapA strains, lacking all known purine transporters, and confers resistance to the antifungal 5-fluorocytosine. Kinetic analyses showed FcyB is a low-capacity, high affinity, cytosine-purine transporter sharing similar molecular interactions for substrate recognition with the yeast Fcy2p/Fcy21p carriers. fcyB transcription is highly activated during germination but drops at low constitutive levels throughout vegetative development. UaY-mediated purine induction of fcyB transcription is only moderate, while ammonium represses transcription through an AreA-dependent mechanism. A strain expressing FcyB-GFP confirms a low protein expression level in the plasma membrane of vegetative mycelia, but reveals an abundant expression in sexual and asexual compartments. FcyB-GFP was also shown to be downregulated by endocytosis in response to ammonia or the presence of cytosine. The expression profile of FcyB supports that its main physiological role is cytosine-purine scavenging. PMID- 18384519 TI - Eight-week regimen of antiviral combination therapy with peginterferon and ribavirin for patients with chronic hepatitis C with hepatitis C virus genotype 2 and a rapid virological response. AB - BACKGROUND: It remains unclear how we can shorten the treatment duration of antiviral combination therapy with peginterferon and ribavirin for patients with chronic hepatitis C virus (HCV) genotype 2 infection who achieved a rapid virological response (RVR). AIM: We compared the efficacy of antiviral combination therapy with peginterferon and ribavirin for 8 vs. 24 weeks for the treatment of patients with HCV genotype 2 infection and with RVR. METHODS: Sixty one patients were enrolled. Serum HCV RNA was not detected at 4 weeks after the start of treatment in 32 patients with an RVR. These 32 patients were randomly assigned to 8-week (n=15) or 24-week (n=17) treatment regimens. Patients in the 8 week group who relapsed underwent a 24-week retreatment. RESULTS: No significant difference in patient characteristics was observed between the 8- and the 24-week treatment groups. A sustained virological response (SVR) was seen in five of 15 patients (33.3%) in the 8-week treatment group and 14 of 17 (82.4%) in the 24 week treatment group; the rate was significantly higher in the 24-week treatment group (P=0.0140). Nine of 10 relapsed patients in the 8-week treatment group underwent a 24-week retreatment, and seven achieved an SVR. CONCLUSION: An 8-week regimen of combination antiviral therapy with peginterferon and ribavirin yielded an increase in the relapse rate, indicating the limitation of a reduction of treatment below 12 weeks in patients with genotype 2, after RVR. PMID- 18384520 TI - Is serum alanine transaminase level a reliable marker of histological disease in chronic hepatitis C infection? AB - BACKGROUND: Approximately 20-30% of patients chronically infected with hepatitis C virus (HCV) have persistently normal alanine transaminase (PNALT) levels. These patients are described to have a mild degree of histological liver damage. We aimed to assess the histological liver changes in HCV patients with PNALT. PATIENTS AND METHODS: Sixty-five patients with HCV and PNALT (group A) underwent a liver biopsy. PNALT was defined as three or more determinations identified to be within the normal range over 6 months or longer. The demographical features and histological changes were compared with 66 consecutive patients with chronic HCV infection and elevated ALT (group B). All patients had a detectable HCV RNA. Histological disease was scored according to the METAVIR system. RESULTS: Females were more likely to have normal ALT levels (65%). The mean ALT level in Group A and B was 30 and 105 IU/L respectively. No patient in either group had normal histology. The mean necro-inflammatory scores in groups A and B (2.0+/-0.68 vs 2.09+/-0.67) and the mean fibrosis scores (2.11+/-0.87 vs 2.24+/-1.04) were not significantly different. Bridging fibrosis in groups A and B was seen in 24.6 and 37.9% patients, respectively, while cirrhosis was seen in 6.2 and 7.6% patients respectively. Hepatic steatosis in groups A and B (0.94+/-0.86 vs 1.0+/-1.02 respectively) was also not significantly different and did not show any association with the fibrosis scores across the two groups. In group A, the necro inflammatory and fibrosis scores of patients with and without steatosis were not statistically significant. Age was the only predictor of normal ALT levels. However, increasing age did not show a significant increase in histological activity in either group beyond a certain age. CONCLUSION: This study demonstrates that ALT is a poor surrogate marker for inflammation and fibrosis in HCV patients. Given the presence of significant necro-inflammation in PNALT patients, the risk/benefit ratio justifies treatment without the need for a liver biopsy. PMID- 18384521 TI - Prevalence and associated factors of non-alcoholic fatty liver disease in patients with type-2 diabetes mellitus. AB - BACKGROUND/AIMS: Diabetic patients have an increased prevalence and severity of non-alcoholic fatty liver disease (NAFLD). We aimed to investigate the prevalence and the factors associated with the presence of ultrasonographic NAFLD in type-2 diabetic individuals. METHODS: In a cross-sectional design study, 180 type-2 diabetic patients were submitted to a complete clinical and laboratory evaluation and abdominal ultrasonography for NAFLD detection and grading. Statistical analysis included bivariate tests, analysis of variance (anova, for increasing severity of steatosis) and multivariate logistic regression. RESULTS: The prevalence of ultrasonographic NAFLD was 69.4% [95% confidence interval (CI): 58.3-82.7%]. Patients with NAFLD were more obese, had a higher waist circumference and serum triglyceride and alanine aminotransferase (ALT) levels than those without steatosis. Neither diabetic degenerative complication, nor glycaemic control was associated with liver steatosis. On multivariate analysis, a high serum triglycerides level [>2.82 mmol/L, odds ratio (OR): 3.7-4.1, 95% CI: 1.2-13.3] and a high-normal ALT level (> or =40 U/L, OR: 2.5-2.7, 95% CI: 1.2 5.9) were independently associated with hepatic steatosis, together with either the presence of obesity (OR: 7.1, 95% CI: 3.0-17.0) or of increased waist circumference (OR: 4.8, 95% CI: 1.9-12.2). CONCLUSIONS: Type-2 diabetic patients have a high prevalence of ultrasonographic NAFLD and its presence is associated with obesity, mainly abdominal, hypertriglyceridaemia and high-normal ALT levels. Non-alcoholic fatty liver disease in diabetic patients may develop and progress independent of the diabetes progression itself. PMID- 18384522 TI - Role of nitric oxide synthesized by nitric oxide synthase 2 in liver regeneration. AB - BACKGROUND/AIMS: Nitric oxide synthase 2 (NOS2) is expressed during liver regeneration after a partial hepatectomy (PHx); NOS2 subsequently synthesizes nitric oxide (NO). However, the role of NOS2-synthesized NO in post-PHx liver regeneration remains unclear. We investigated the role of NOS2-synthesized NO in liver regeneration. METHODS: NOS2 knockout (NOS2-KO) mice and control mice were subjected to PHx. Liver mass recovery and serum alanine aminotransferase (ALT) levels were then evaluated. The expressions of Ki-67 and single-strand DNA were also evaluated in remnant liver specimens. Differences in the gene expression profiles of the two groups of remnant liver specimens were analysed using a microarray and were validated using a reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In NOS2-KO mice, liver regeneration was delayed and apoptosis and serum ALT levels were higher than the levels in the control mice. A microarray study and RT-PCR revealed that heat shock protein 70 family (HSP70 family), haeme oxygenase 1 (Hmox1), neuropilin 1 (Nrp1) and epidermal growth factor receptor (EGFR) were downregulated in NOS2-KO mice. CONCLUSIONS: NOS2 synthesized NO may improve hepatocyte viability through the induction of the HSP70 family and Hmox1 and may sensitize the remnant liver to growth factors through the induction of Nrp1 and EGFR post-PHx. PMID- 18384523 TI - Hepatocellular carcinoma incidence trends in Canada: analysis by birth cohort and period of diagnosis. AB - BACKGROUND AND AIMS: We examined birth cohort and calendar period trends in hepatocellular carcinoma (HCC) incidence in Canada (1976-2000). We also projected HCC incidence rates through 2015. PATIENTS AND METHODS: Data were obtained from the Canadian Cancer Registry on all cases of HCC diagnosed among persons aged 20 years and older in Canada from 1976 to 2000 and was used to describe trends in HCC incidence rates. RESULTS: We found that age-adjusted HCC incidence rates increased faster in males compared with females, 3.4% per year [95% confidence interval (CI): 3.0-3.8%] vs 2.2% per year (95% CI: 1.5-2.8%). An increasing birth cohort trend accelerated among males around the 1940 birth cohort and decelerated among females around the 1935 birth cohort. For calendar period trends, the increasing HCC risk was relatively constant over time among males whereas there was an acceleration in HCC risk around 1988 among females. Age-adjusted HCC incidence rates were projected to increase 73% in males and 28% in females from 1996 to 2015. CONCLUSIONS: Our results suggest that HCC incidence rates will continue to increase in Canada during the next decade as persons born in more recent birth cohorts, who face a relatively greater risk for HCC, age. PMID- 18384524 TI - Lactic acid production by Lactobacillus delbrueckii in a dual reactor system using packed bed biofilm reactor. AB - AIMS: An integrated dual reactor system for continuous production of lactic acid by Lactobacillus delbrueckii using biofilms developed on reticulated polyurethane foam (PUF) is demonstrated. METHODS AND RESULTS: Lactobacillus delbrueckii was immobilized on PUF, packed in a bioreactor and used in lactic acid fermentation. The rate of lactic acid production was significantly high with a volumetric productivity of 5 g l(-1) h(-1) over extended period of time. When coupled to a bioreactor, the system could be operated as dual reactor for over 1000 h continuously without augmentation of inoculum and no compromise on productivity. CONCLUSIONS: Polyurethane foams offer an excellent support for biofilm formation. SIGNIFICANCE AND IMPACT OF THE STUDY: The system was very robust and could be operated for prolonged period at a volumetric productivity of 4-6 g l(-1) h(-1). PMID- 18384525 TI - Optimization of a new heteropolysaccharide production by a native isolate of Leuconostoc sp. CFR-2181. AB - AIM: This work is aimed at optimizing the production of a new heteropolysaccharide (HePS) of Leuconostoc sp. CFR-2181 by standardizing the fermentation conditions in a low cost semi-synthetic medium. METHODS AND RESULTS: Both nutritional and cultural parameters, such as carbon source and its concentration, initial pH of the exopolysaccharide (EPS) medium, fermentation temperature and fermentation period were optimized. Fermentation of the EPS medium (pH 6.7), containing sucrose at 5% (w/v) and 5% (v/v) inoculum, at 25 degrees C resulted in maximum production of HePS (18.38 g l(-1)) by the isolate in 4 h of fermentation. CONCLUSIONS: The isolate was found to produce good amount of HePS in just 4 h in a low cost semi-synthetic EPS medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on rapid production of HePS by any lactic culture, which can significantly reduce the cost of the EPS. PMID- 18384526 TI - Impact of topological and electronic descriptors in the QSAR of pyrazine containing thiazolines and thiazolidinones as antitubercular and antibacterial agents. AB - Quantitative structure activity relationships (QSAR) were developed relating the 14th and 21st day antituberculosis activity against H(37)Rv strain of Mycobacterium tuberculi and antibacterial activity against Staphylococcus aureus ATCC3750, Bacillus subtilis 6633, Escherichia coli ATCC3750, and Salmonella typhi NCTC786 of 55 pyrazine containing thiazolines and thiazolidinones, with the molecular descriptors. The developed models were able to fit the data well (r(2) = 0.69-0.87) and had reasonable predictive capability (q(2) > 0.62). The data were also divided into a training set and a test set, the former was used to develop the QSAR and the latter was used to evaluate the predictive capability of these developed models. In all the cases, the models were able to predict the test data set reasonably well. Predominantly, pyrazine ring is well-known for its antimycobacterial activity and hence these equation could be used to design newer analogues with higher activity. These compounds also possess both antitubercular and antibacterial activity. Descriptors pertaining to electronic, topology, and hydrophobicity of the molecules appear in the model equations. PMID- 18384527 TI - A theoretical study on interactions between mitoxantrone as an anticancer drug and DNA: application in drug design. AB - This research is an effort to further understand the physicochemical interaction between the novel drug, mitoxantrone (MTX) and its biologic receptor, DNA. The ultimate goal is to design drugs that interact more with DNA. Understanding the physicochemical properties of the drug as well as the mechanism by which it interacts with DNA, it should ultimately allow the rational design of novel anti cancer or anti-viral drugs. Molecular modelling on the complex formed between MTX and DNA presented that this complex was indeed fully capable of participating in the formation of a stable intercalation site. Furthermore, the molecular geometries of MTX and the DNA bases (adenine, guanine, cytosine and thymine) were optimized with the aid of the B3LYP/6-31G* method. The properties of the isolated intercalator and its stacking interactions with the adenine...thymine (AT) and guanine...cytosine (GC) nucleic acid base pairs were studied with the DFTB method (density functional tight-binding), an approximate version of the DFT method, that was extended to cover the London dispersion energy. The B3LYP/6-31G* stabilization energies of the intercalator...base pair complexes were found 10.06 kcal/mol and 21.64 kcal/mol for AT...MTX and GC...MTX, respectively. It was concluded that the dispersion energy and the electrostatic interaction contributed to the stability of the intercalator.DNA base pair complexes. The results concluded from the comparison of the DFTB method and the Hartree-fock method point out that these methods show close results and support each other. PMID- 18384528 TI - Transformational leadership in drug discovery by way of virtuous thought, word and deed. PMID- 18384529 TI - Novel method for probing the specificity binding profile of ligands: applications to HIV protease. AB - A detailed understanding of factors influencing the binding specificity of a ligand to a set of desirable targets and undesirable decoys is a key step in the design of potent and selective therapeutics. We have developed a general method for optimizing binding specificity in ligand-receptor complexes based on the theory of electrostatic charge optimization. This methodology can be used to tune the binding of a ligand to a panel of potential targets and decoys, along the continuum from narrow binding to only one partner to broad binding to the entire panel. Using HIV-1 protease as a model system, we probe specificity in three distinct ways. First, we probe interactions that could make the promiscuous protease inhibitor pepstatin more selective toward HIV-1 protease. Next, we study clinically approved HIV-1 protease inhibitors and probe ways to broaden the binding profiles toward both wild-type HIV-1 protease and drug-resistant mutants. Finally, we study a conformational ensemble of wild-type HIV-1 protease to 'design in' broad specificity to known drugs before resistance mutations arise. The results from this conformational ensemble were similar to those from the drug resistant ensemble, suggesting the use of a conformational wild-type ensemble as a tool to develop escape-mutant-resistant inhibitors. PMID- 18384531 TI - Can disturbances in the atmospheric electric field created by powerline corona ions disrupt melatonin production in the pineal gland? AB - Recent epidemiological studies have reported an increased risk of leukemia in adults and children near overhead high voltage powerlines at distances beyond the measured range of the direct electric and magnetic fields. Corona ions are emitted by powerlines, forming a plume that is carried away from the line by the wind. The plume generates highly variable disturbances in the atmospheric electric field of tens to a few hundred V/m on time scales from seconds to minutes. Such disturbances can be seen up to several hundred meters from powerlines. It is hypothesized that these random disturbances result in the disruption of nocturnal melatonin synthesis and related circadian rhythms, in turn leading to increased risk of a number of adverse health effects including leukemia. In support of the hypothesis, it is noted that melatonin is highly protective of oxidative damage to the human hemopoietic system. A review of electric field studies provides evidence that (i) diurnal variation in the natural atmospheric electric field may itself act as a weak Zeitgeber; (ii) melatonin disruption by electric fields occurs in rats; (iii) in humans, disturbances in circadian rhythms have been observed with artificial fields as low at 2.5 V/m. Specific suggestions are made to test the aspects of the hypothesis. PMID- 18384530 TI - Critical role of glutathione in melatonin enhancement of tumor necrosis factor and ionizing radiation-induced apoptosis in prostate cancer cells in vitro. AB - The role of antioxidants in reducing cancer initiation and progression has been highlighted in recent years. Not only antioxidants limit cancer cell growth but also, in some situations, they promote the effectiveness of conventional treatments. Melatonin, an endogenously synthesized antioxidant, reduces cell growth of several tumor types both in vivo and in vitro. Additionally, the indole limits the collateral damage induced by many chemotherapeutic agents. By using a cellular model of human prostate cancer, we studied the ability of melatonin to enhance apoptosis induced by tumor necrosis factor or gamma radiation. It has been reported that melatonin reduces prostate cancer cell growth and, more recently, it promotes cell differentiation. In this work, we also show that melatonin elevates p21 protein levels and increases antioxidant capacity of prostate cancer cells. In addition, melatonin significantly enhances hrTNFalpha induced cell death by decreasing NFkappaB activation. Bcl-2 and survivin down regulation appears to be associated to apoptosis stimulation under NFkappaB inhibition. On the contrary, melatonin does not promote irradiation-induced cell death due to an increment in intracellular glutathione content. In conclusion, prevention of NFkappaB activation by melatonin enhances the effectiveness of cytokine treatment in prostate cancer cells but it is not sufficient to enhance cell death triggered by other therapies which generate free radicals. A crucial role of glutathione in survival mechanisms of prostate cancer cells should be carefully considered. PMID- 18384532 TI - Regulatory influence of melatonin on collagen accumulation in the infarcted heart scar. AB - The regulatory influence of the pineal gland on superficial wound healing and collagen content is documented. The aim of the present study was to determine whether the pineal gland and its secretory product melatonin regulate collagen accumulation in the scar of the infarcted heart and to explain the mechanisms of its action. To induce myocardial infarction in rats the left coronary artery was ligated. Metoprolol at the dose of 0.2 mg/100 g body weight (b.w.) was injected intraperitoneally to inhibit melatonin secretion. Pinealectomy was performed on some animals. For the in vitro study, cells were isolated from the heart scar and cultured in Dulbecco's modified Eagle medium with 3% fetal calf serum and antibiotics. Collagen content was evaluated as hydroxyproline content according to the Woessner method. Melatonin subcutaneously injected into the rats at the doses of 30 microg/100 g or 60 microg/100 g b.w. increased collagen accumulation in the heart scar. The doses of 3 microg/100 g b.w. and 300 microg/100 g b.w. were not effective. Surgical and pharmacological pinealectomies had opposite effects and reduced collagen content in the scar. However, melatonin administration (60 microg/100 g b.w.) to pinealectomized rats reversed the effect of pinealectomy and normalized collagen levels in heart after infarction. Cells isolated from the heart scar were identified as myofibroblasts. Melatonin (10(-7) 10(-8) m) increased collagen accumulation in the cultures. Collagen accumulation in the scar of the infarcted heart is regulated by melatonin and it exerts effects directly on the myofibroblasts of the infarcted area. Therefore, melatonin-induced collagen accumulation in the infarcted heart could be considered as the event improving the tensile strength of the scar and retarding the development of complications. PMID- 18384533 TI - Fluoride content in table salt distributed in Mexico City, Mexico. AB - OBJECTIVE: The aim of this study was to analyze table salt available in Mexico City's market to identify the fluoride concentrations and to compare these with the Mexican regulations. METHODS: We analyzed 44 different brands of table salt. All samples were purchased at random in different stores, supermarkets, and groceries from Mexico City's metropolitan area and analyzed in triplicate in three different laboratories (nine determinations per sample) with an Orion 720 A potentiometer and an Orion 9609 BN ion-specific electrode. RESULTS: Fluoride concentration in the samples varied from 0 ppm to 485 ppm. It was found that fluoride concentration varied widely among the analyzed brands. Also, we found that fluoride concentration in 92 percent of the analyzed samples did not match with that printed on the label. Only 6.8 percent of the analyzed samples contained fluoride concentrations that meet Mexican and WHO regulations. CONCLUSIONS: The broad variation in the analyzed samples suggests that Mexican Public Health authorities must implement more stringent regulation guidelines and procedures for controlling the distribution of salt and its fluoride concentration for human consumption. PMID- 18384534 TI - Obesity and dental caries in children aged 2-6 years in the United States: National Health and Nutrition Examination Survey 1999-2002. AB - OBJECTIVE: This study assessed the associations between obesity and dental caries in young children participating in a national survey. METHODS: Participants included 1507 children aged 2-6 years who received dental examinations and had at least 10 primary teeth in the National Health and Nutrition Examination Survey 1999-2002. Decayed/filled teeth (dft) counts of primary dentition were obtained, and weight and height were measured. Body mass index (BMI; kg/m2) was calculated, and participants were categorized using age- and gender-specific criteria as underweight (<5th percent), normal (5th-85th percent), at risk for overweight (>85th and <95th percent), and overweight (> or =95th percent). With appropriate sample weighting, relationships between dft and BMI were assessed using the Kruskal-Wallis test and multivariable logistic regression. RESULTS: Seventy-four percent of children were classified as normal weight, 11 percent as at risk for overweight, and 11 percent as overweight; 58 percent did not have caries; 30 percent had 1-5 dft and 12 percent had >5 dft. When caries experience was compared across BMI categories stratified by age and race characteristics, statistically significant association between caries and obesity was found only for 60- <72-month age group. In the comparison between children with normal and at-risk BMI only, significant associations were also found in the Hispanic and non-Hispanic Black strata. In multivariable logistic regression models to predict caries experience, family income and age were statistically significant predictors for severe early childhood caries only. CONCLUSIONS: There appears to be no significant association between childhood obesity and caries experience after controlling forage, race, and poverty/income ratio. However, further studies are needed to better understand this relationship. PMID- 18384535 TI - The potential role of breast-feeding and other factors in helping to reduce early childhood caries. AB - OBJECTIVES: Dental caries is the most common chronic disease in US children. Early childhood caries (ECC) is particularly virulent and can interfere with a child's ability to eat, grow, speak, and communicate. Studies on whether breast feeding or bottle-feeding are more likely to reduce ECC have proven inconclusive. METHODS: The study population included 175 children, aged 1 to 5, receiving dental care at the Hughes Spalding Children's Hospital in Atlanta, GA. Participation included a dental exam, chart data abstraction, and a personal interview with the mother. RESULTS: Too few exclusively breast-fed children prevented the adequate study of breast-feeding. However, children exclusively bottle-fed for at least 1.5 years had more decayed or filled tooth surfaces than children breast-fed part of that time but well short of a year. No bottle at night nor juice at irregular times, the mother's brushing of her child's teeth, and adequate dental care in the mother seemed to reduce ECC. CONCLUSIONS: Our results suggest measures that might reduce ECC risk. Medical providers must discuss oral health with new mothers and educate them on the important role they play in keeping their babies' teeth healthy PMID- 18384536 TI - A pharmacokinetic study of enrofloxacin and its active metabolite ciprofloxacin after oral and intramuscular dosing of enrofloxacin in rhesus monkeys (Macaca mulatta). AB - Enrofloxacin is used for treating Shigellosis in non-human primates; however, there are no reports describing its pharmacokinetics in rhesus monkeys. Pharmacokinetic data in intended target species (rhesus) help to determine the proper dose regimen. Blood levels of enrofloxacin and ciprofloxacin (enrofloxacin's active metabolite), were determined after either intramuscular or oral dosing of enrofloxacin for 7 days in a cross-over study. Levels of both antibiotics were determined by solid phase extraction followed by reversed-phase chromatography with tandem mass spectrometry. Results indicate enrofloxacin half life after intramuscular dosing is estimated to be 2.4 hours. Enrofloxacin given either intramuscular or p.o. rapidly achieves satisfactory therapeutic blood levels of enrofloxacin or ciprofloxacin in rhesus monkeys. Results from these pharmacokinetic study parallel values published for other animal species. Our results show use of enrofloxacin is effective in managing Shigella infections in rhesus monkeys based upon achieving these blood drug levels. PMID- 18384537 TI - Behavioural flexibility in individuals with Angelman syndrome, Down syndrome, non specific intellectual disability and Autism spectrum disorder. AB - BACKGROUND: Little is known about behavioural flexibility in children and adults with Angelman syndrome and whether people with this syndrome have more or less problems in being behaviourally flexible as compared with other people. METHOD: Behavioural flexibility scores were assessed in 129 individuals with Angelman syndrome using 11 items from the Behavioural Flexibility Rating Scale-Revised (Green et al. 2007). Level of behavioural flexibility scores in individuals with Angelman syndrome (N = 129) was compared with that of people with non-specific intellectual disability (ID) (N = 90), Down syndrome (N = 398) and Autism spectrum disorder (N = 235). RESULTS: Comparative analyses show that individuals with Angelman syndrome were more flexible than those with non-specific ID (P < 0.001) and those with Autism spectrum disorder (P < 0.01). There were no differences in behavioural flexibility scores between individuals with Angelman syndrome and those with Down syndrome (P = 0.94). CONCLUSION: It is concluded that individuals with Angelman syndrome are comparatively flexible in their behaviour. PMID- 18384538 TI - Molecular phylogeny and divergence dates for Australasian elapids and sea snakes (hydrophiinae): evidence from seven genes for rapid evolutionary radiations. AB - One of the most prolific radiations of venomous snakes, the Australo-Melanesian Hydrophiinae includes approximately 100 species of Australasian terrestrial elapids plus all approximately 60 species of viviparous sea snakes. Here, we estimate hydrophiine relationships based on a large data set comprising 5800 bp drawn from seven genes (mitochondrial: ND4, cytb, 12S, 16S; nuclear: rag1, cmos, myh). These data were analysed using parsimony, likelihood and Bayesian methods to better resolve hydrophiine phylogeny and provide a timescale for the terrestrial and marine radiations. Among oviparous forms, Cacophis, Furina and Demansia are basal to other Australian elapids (core oxyuranines). The Melanesian Toxicocalamus and Aspidomorphus group with Demansia, indicating multiple dispersal events between New Guinea and Australia. Oxyuranus and Pseudonaja form a robust clade. The small burrowing taxa form two separate clades, one consisting of Vermicella and Neelaps calanotus, and the other including Simoselaps, Brachyurophis and Neelaps bimaculatus. The viviparous terrestrial elapids form three separate groups: Acanthophis, the Rhinoplocephalus group and the Notechis Hemiaspis group. True sea snakes (Hydrophiini) are robustly united with the Notechis-Hemiaspis group. Many of the retrieved groupings are consistent with previous molecular and morphological analyses, but the polyphyly of the viviparous and burrowing groups, and of Neelaps, are novel results. Bayesian relaxed clock analyses indicate very recent divergences: the approximately 160 species of the core Australian radiation (including sea snakes) arose within the last 10 Myr, with most inter-generic splits dating to between 10 and 6 Ma. The Hydrophis sea snake lineage is an exceptionally rapid radiation, with > 40 species evolving within the last 5 Myr. PMID- 18384539 TI - Head shape evolution in Tropidurinae lizards: does locomotion constrain diet? AB - Different components of complex integrated systems may be specialized for different functions, and thus the selective pressures acting on the system as a whole may be conflicting and can ultimately constrain organismal performance and evolution. The vertebrate cranial system is one of the most striking examples of a complex system with several possible functions, being associated to activities as different as locomotion, prey capture, display and defensive behaviours. Therefore, selective pressures on the cranial system as a whole are possibly complex and may be conflicting. The present study focuses on the influence of potentially conflicting selective pressures (diet vs. locomotion) on the evolution of head shape in Tropidurinae lizards. For example, the expected adaptations leading to flat heads and bodies in species living on vertical structures may conflict with the need for improved bite performance associated with the inclusion of hard or tough prey into the diet, a common phenomenon in Tropidurinae lizards. Body size and six variables describing head shape were quantified in preserved specimens of 23 species, and information on diet and substrate usage was obtained from the literature. No phylogenetic signal was observed in the morphological data at any branch length tested, suggesting adaptive evolution of head shape in Tropidurinae. This pattern was confirmed by both factor analysis and independent contrast analysis, which suggested adaptive co-variation between the head shape and the inclusion of hard prey into the diet. In contrast to our expectations, habitat use did not constrain or drive head shape evolution in the group. PMID- 18384540 TI - Generalized granuloma annulare--response to doxycycline. PMID- 18384541 TI - Presence of residual basal cell carcinoma in re-excised specimens is more probable when deep and lateral margins were positive. AB - In order to quantify presence of residual BCC in patients with histologic positive margins after the first excision and to correlate the presence of residual tumor in re-excised lesions with the location of the positive margin on the first excision, a retrospective evaluation of 2053 surgically treated BCC was performed. Only 38.3% of the re-excised lesions showed residual tumor. In the group of re-excised lesions where residual BCC was found, 13% had lateral positive margin in the first excision, 39% had deep positive margin and 48% had both lateral and deep positive margins. In the group of re-excised lesions where no residual BCC was found, 49% of the primary excised lesions had lateral positive margin, 32% had deep positive margin and 19% had both deep and lateral positive margins. The association between residual tumor and positive margins was statistically significant (p = 0.01). Our findings confirm that presence of residual tumour is more likely when both lateral and deep margins are compromised. PMID- 18384542 TI - Alan Lyell and Lyell's syndrome. AB - Alan Lyell has done a major contribution to the description of several diseases which could be Lyell's diseases if he had not been the most unpretentious gentelman in the field of dermatology. PMID- 18384543 TI - An open-label pilot study of alefacept for the treatment of pyoderma gangrenosum. AB - BACKGROUND: Pyoderma gangrenosum (PG) is a chronic inflammatory disease that causes painful cutaneous ulcers that are difficult to treat. Currently, systemic immunosuppressants, often including prednisone, are the mainstay of therapy. Long term therapy with these agents is often required which exposes patients to possible adverse effects. An alternative treatment that is safe and effective is truly needed. OBJECTIVE: To study the efficacy and safety of alefacept, which inhibits T-cell activation and selectively reduces the T-cell population, for treatment of PG. METHOD: In this prospective open-label pilot study, four patients diagnosed with PG received weekly doses of 15 mg alefacept intramuscularly for 20 weeks with 12-week treatment-free follow-up. The primary efficacy end point was the proportion of patients achieving remission as defined by a Physician Global Assessment (PGA) of 'clear' or 'almost clear.' Secondary endpoints included proportion of patients achieving 50% improvement in PG lesion size (measured in mm) and proportion of patients achieving resolution of inflammation (an erythema score of 0 and a border thickness of 0 on scales of 0 4). RESULTS: By week 20, one (25%) of the four patients achieved remission, two showed marked improvement in severity on PGA, and one had slight improvement. One patient showed a 98% decrease in lesion size; two other patients evidenced a decrease in the number of small lesions as well as improvements in primary lesion sizes, but did not surpass the 50% criterion. All four patients showed improved erythema scores during treatment, though only one patient showed a complete resolution of inflammation. LIMITATIONS: It may be difficult to generalize the results of this study to a larger population of patients with PG due to the small sample size and lack of a control group. A longer treatment interval might have been required. Safety and efficacy of long-term therapy is unknown. CONCLUSION: In this pilot study it appears that alefacept treatment may significantly reduce PG severity levels as evidenced by improvement in PGA, Subject Global Assessment, and inflammation scores in all patients. Alefacept may be a safe and effective alternative to current systemic immunosuppressants used to treat PG. Double blinded, controlled trials are necessary to further evaluate the safety and effectiveness of this treatment. PMID- 18384544 TI - Cogent differences in skin heme oxygenase-1 levels between psoriasis patients and healthy controls. PMID- 18384545 TI - Tattoo-induced vasculitis. PMID- 18384546 TI - Commentary on: 'Molluscum contagiosum on region of burned skin'. PMID- 18384547 TI - Re: Baum S, Scope A, Barzilai A, Azizi E, Trau H. The role of IVIg treatment in severe pemphigus vulgaris. J Eur Acad Dermatol Venereol 2006; 20: 548-52. PMID- 18384548 TI - Sun protection among skin cancer-treated patients. PMID- 18384549 TI - Recurrent Scedosporium apiospermum skin infection in a renal transplant recipient. PMID- 18384550 TI - Rifampicin in the treatment of psoriasis. PMID- 18384551 TI - Treatment of classic Kaposi sarcoma with a nicotine dermal patch: a phase II clinical trial. AB - BACKGROUND: Kaposi sarcoma (KS), a malignancy of dermal endothelial cells that is caused by human herpesvirus 8 (HHV8) infection, is sensitive to perturbations of immunity. Nicotine might be effective against KS because of its immunologic and vascular effects and because smoking is associated with a low risk of KS. OBJECTIVE AND STUDY DESIGN: We conducted a masked, randomized phase 2 clinical trial of transdermal nicotine and placebo patches to assess the safety and efficacy of nicotine against classic KS (cKS). SUBJECTS AND METHODS: Three cKS lesions, predominantly nodules, in each of 24 non-smoking patients were randomly assigned to 15 weeks continuous treatment with nicotine patch (escalated to 7 mg), identical masked placebo patch or no patch. Changes in lesion area and elevation from baseline through six follow-up visits, by direct measurement and by two independent readers using digital photographs of the lesions, were compared using non-parametric and regression methods. Changes in longitudinal levels of HHV8 antibodies and DNA in blood cells were similarly assessed. RESULTS: There were no systemic or serious adverse events, and compliance was good. One patient resumed smoking and discontinued patches, and two patients withdrew at week 12 for unrelated indications. Six (29%) of the remaining 21 suspended use of patches to relieve local skin irritation; four of these six completed the trial at reduced dose. Treatment assignment was not associated with significant or consistent changes in cKS lesion area or elevation, HHV8 viral load or antibodies. CONCLUSION: Transdermal nicotine and placebo patches caused no serious toxicities but had no demonstrable effect on nodular cKS lesions or HHV8 levels. PMID- 18384553 TI - Serologic markers of celiac disease in psoriatic patients. AB - BACKGROUND: Aetiopathogenesis of psoriasis is complex and not yet well known. In recent years, it has been observed that psoriasis can coexist with clinically asymptomatic celiac disease and a gluten-free diet helps to obtain remission, even in patients with very chronic lesions. OBJECTIVE: The aim of our work was to investigate how often the positive titres of antibodies characteristic for celiac disease occur in psoriatics' serum in exacerbation in comparison with controls. PATIENTS/METHODS: Serum samples from 67 patients with intensified psoriatic lesions were investigated. Serum from healthy people at a comparable age and with no familial predisposition to psoriasis and celiac disease was the control material. Antibodies against human tissue transglutaminase (recombinant antigen), against tissue transglutaminase isolated from guinea pig's liver and against gliadin were determined by enzyme-linked immunosorbent assay technique. Anti endomysial antibodies were determined by indirect immunofluorescence method. RESULTS: Patients with psoriasis have significantly higher mean concentrations of antibodies against tissue transglutaminase (human recombinant and guinea pig derived antigen) and against gliadin for IgA. IgA antibodies against tissue transglutaminase (both antigens) and gliadin positively correlate with psoriasis activity. No anti-endomysial antibodies for IgA were found in any serum. CONCLUSIONS: Our results seem to imply an association between psoriasis and asymptomatic celiac disease/gluten intolerance. High percentage of positive results to guinea pig-derived tTG could be due to cellular activity of tissue transglutaminase in psoriasis. PMID- 18384552 TI - A randomized, double-blind study to assess the efficacy of addition of tetracycline to triamcinolone acetonide in the treatment of moderate to severe atopic dermatitis. AB - OBJECTIVE: To assess the efficacy of tetracycline in triamcinolone acetonide ointment compared with triamcinolone acetonide ointment in patients with moderate to severe atopic dermatitis. DESIGN: Randomised, double-blind parallel group study of 8 weeks' duration. SETTING: Outpatient clinic in a university hospital. PARTICIPANTS: Forty-four adult patients with moderate to severe atopic dermatitis (objective SCORAD > 25). INTERVENTIONS: Initial phase (2 weeks): 3% tetracycline 0.1% triamcinolone acetonide vs. 0.1% triamcinolone acetonide twice daily all over the body. MAINTENANCE PHASE: (6 weeks) 0.1% triamcinolone acetonide once daily for 2 weeks, followed by every other day for 2 weeks. In the last 2 weeks, two applications a week were done. An emollient was used additionally once daily. MAIN OUTCOME MEASURES: Primary outcomes were the disease severity scores assessed by objective SCORAD and SASSAD at week 2. Secondary outcomes were the objective SCORAD and SASSAD at weeks 4 and 8, and Staphylococcus aureus colonization at weeks 0 and 2. RESULTS: No significant differences in disease severity outcomes were found between the two groups. Both groups showed clinically relevant improvements in disease severity compared with baseline at weeks 2 and 4. At week 8, there was some worsening in disease severity in both groups, but the disease severity was still significantly lower than at the beginning of the study. Improvement of bacterial colonization was seen in 14 (63.6%) out of the 22 patients in the 3% tetracycline 0.1% triamcinolone acetonide group and in 5 (22.7%) out of the 22 patients in the 0.1% triamcinolone acetonide group. CONCLUSION: The addition of tetracycline was effective on skin colonization by S. aureus but did in our patients with atopic dermatitis not result in a significantly different improvement compared with the group treated without tetracycline. PMID- 18384554 TI - Quality of life in Polish patients with psoriasis. PMID- 18384555 TI - Ano-genital granulomatosis treated with ciprofloxacin. PMID- 18384556 TI - Sparing of the bulge area could preserve intact lower portion of hair follicles in a case of tufted folliculitis. PMID- 18384557 TI - Imatinib-associated hyperpigmentation, a side effect that should be recognized. PMID- 18384558 TI - Aggressive intermediate cell histiocytosis successfully treated by 2 chlorodeoxyadenosine. PMID- 18384559 TI - Alopecia areata in Turkey: demographic and clinical features. AB - BACKGROUND: Alopecia areata is a complex genetic disease with still many unknown aspects, and many studies have been tried to find some clues about it. OBJECTIVE: We aimed to investigate the demographic and clinical characteristics of alopecia areata in Turkish patients. METHODS: Demographic data, localization, attack number in addition to some parameters such as disease duration, severity, age of onset, family history and ophiasis pattern were evaluated in 539 alopecia areata patients. RESULTS: The male to female ratio was 1.6:1. Occipital and beard moustache areas were mostly affected. Positive family history was noticed in 24.1% of the patients. The age of onset was earlier in women than in men (P = 0.04). Severe forms showed more persistent (> or = 1 year) disease duration (P = 0.00). Ophiasis was more common in severe, long duration (> or = 1 year) and early onset (< or = 18 years) disease (P = 0.00 for all parameters). Childhood alopecia areata (< or = 18 years) was also associated with long duration of the disease (P = 0.016) and positive family history (P = 0.008) when compared with adult onset (> 18 years) alopecia areata. PMID- 18384560 TI - Opposed bilateral transposition flap: a simple and effective way to close large defects, especially of the limbs. AB - BACKGROUND: Excision of large tumours, particularly of the limbs, can be challenging because of problems related to wound repair. This is especially true of the lower legs, where skin is often tight and difficult to mobilize. Closure by flap, which would represent the first choice for defects usually between 12-15 mm to 38-40 mm diameter, is at risk of developing complications, such as end-flap necrosis or dehiscence due to skin tension. For larger defects, usually more than 40 to 45 mm diameter, grafting still remains the only realistic option in the majority of cases, with all the various problems associated with this procedure, such as lengthy healing times and the risk of developing leg ulcers, above all in elderly patients with impaired blood circulation. Second intention healing implies extraordinarily long healing times with often unacceptable delays in normal ambulation and activity. OBJECTIVE: To find an alternative to the usual repair techniques and to try to reduce the risk of complications. CONCLUSIONS: We developed a relatively simple but effective technique for the closure of large wounds resulting from the excision of tumours. Our technique consists of two longitudinal, parallel, transposition flaps obtained from two opposite sides of the wound, with major axes orientated in the cephalic-caudal direction. The two flaps are then rotated around two fulcra placed at two extremes of the wound by approximately 90 degrees . This relatively simple technique has never caused any of the ordinarily associated problems in terms of necrosis or ulcer development. In addition, dehiscence of sutures never occurred, given the fact that suture tension is minimal. Quick healing has resulted in the majority of cases, avoiding all the problems associated with grafting or other traditional flap techniques. PMID- 18384561 TI - Mutation screening of entire keratin 5 and keratin 14 genes and identification of a novel mutation in a Chinese family with epidermolysis bullosa simplex Dowling Meara. PMID- 18384562 TI - Autosomal-dominant hypohidrotic ectodermal dysplasia caused by a novel mutation. PMID- 18384563 TI - Statinat-induced pemphigus: over-the-counter drugs too may be harmful! PMID- 18384564 TI - Pseudoscabies caused by Dermanyssus gallinae in Italian city dwellers: a new setting for an old dermatitis. PMID- 18384565 TI - A study on serum IgE and clinical symptomatology of atopy in patients suffering from the lysosomal storage disorder Fabry disease. AB - BACKGROUND: Lysosomal storage disorders may impair intracellular lysosomal processing of antigen with consequences for antibody production [e.g. immunoglobulin E (IgE)] and atopic disease status. AIMS: Serum concentration of total IgE as well as clinical symptoms of atopic disorders as an indirect consequence of lysosomal impairment of antigen processing were studied in male and female Fabry disease (FD) patients with and without replacement of the missing lysosomal enzyme, alpha-galactosidase A (alpha-Gal A). METHODS: Observational study in 31 adult FD patients with measurements of total serum IgE concentration. Questionnaire-derived data were obtained for atopic eczema (AE) skin lesions, allergic rhinoconjunctivitis (RCA) and allergic asthma (AA) at present or in the past. RESULTS: Among 12 FD males under enzyme replacement therapy (ERT), 2 showed total IgE concentrations above 100 kU/L. Clinical symptoms for AA were found in 2, RCA and AE in 1, respectively. Among 10 FD females under ERT, 4 showed total IgE concentrations above 100 kU/L. Clinical symptoms for AA were found in 4, RCA in 2 and AE in 2. Among 9 females without ERT, 2 showed total IgE concentrations above 100 kU/L. Clinical symptoms for AA were found in 2, RCA in 2 and AE in none. CONCLUSIONS: FD patients may demonstrate an increased total serum IgE concentration and may show symptoms of atopic disorders (AA, RCA, AE) in a prevalence rate comparable to international experience for individuals without FD. There was no difference between patients with and without ERT. Lack of detection of AE in females without ERT is suggested to be caused by the small sample size. FD patients without any alpha-Gal A activity prior to initiation of ERT should be in the focus of future studies. PMID- 18384566 TI - Ascending aortic dissection after proximal bypass anastomotic device. AB - Ascending aortic dissection is a known complication of cardiac surgery. Different type of devices allowing performance of proximal bypass anastomoses without aortic side-clamping are supposed to reduce the risk for aortic dissection. We report a case in which ascending aortic dissection occurred six months after off pump coronary artery bypass surgery, with the entry tear arising directly from a proximal anastomosis performed with the Heartstring hemostatic seal system. PMID- 18384567 TI - Expanding the utilization of Contegra for ventricular septal defect repair. AB - Different patch materials have been utilized in repairing ventricular septal defect (VSD) with great success. In this report, in addition to the right ventricular outflow tract reconstruction, the VSD was repaired successfully in all cases by fashioning a patch from a segment of the Contegra xenograft conduit. The freedom from infection, thromboembolism, and reintervention during follow-up, in addition to the advantage of ready availability and cost-effectiveness offered by using the same bovine material, imply that Contegra xenograft is a promising alternative patch material for VSD repair. PMID- 18384568 TI - Coronary artery dissection with rupture of aortic valve commissure following type A aortic dissection: the role of 64-slice MDCT. AB - A rare case of bilateral coronary artery dissection with rupture of aortic valve commissure following type A aortic dissection is described. 64-slice multidetector computed tomography (MDCT) was able to demonstrate both this findings along with involvement of other neck vessels. TEE demonstrated the severity and mechanisms of aortic valve damage and assisted the surgeon in valve repair. MDCT has played an invaluable role in the diagnosis of the abnormal details of such life-threatening vascular complications. PMID- 18384569 TI - A case series of myectomy: a review of its current application as an adjunct procedure. AB - In this article, we review the role played by myectomy in cardiac surgery. For this purpose, we looked at three cases of different etiology where myectomy in conjunction with treatment of the primary condition proved invaluable in the outcome of these patients. The primary conditions requiring treatment were subaortic membrane, hypertrophic obstructive cardiomyopathy with mitral valve regurgitation, and aortic stenosis. PMID- 18384570 TI - A safe option to perform an apparent difficult redo surgery. AB - A 50-year-old female operated of Bentall five years before was referred to our hospital for an aneurysm of both right subclavian artery and brachiocephalic trunk associated with a false anastomotic aneurysm on the insertion of the left coronary artery. The procedure was performed under moderate hypothermic circulatory arrest; the false aneurysm was repaired, the brachiocephalic trunk and the subclavian aneurysm were resected, an aorto-carotid and axillary bypass were finally performed. The postoperative course was uneventful. She was discharged to home on postoperative day 7. At six-month follow-up, she was still asymptomatic. PMID- 18384571 TI - "Gumnut" device for off-pump proximal aortocoronary anastomosis. AB - Use of side-biting clamps in patients with coronary artery disease and atheromatous disease of ascending aorta, undergoing coronary artery surgery can lead to major postoperative neurological events. We describe the use of a simple device to avoid use of side-biting clamp on aorta for proximal aortocoronary anastomosis. PMID- 18384572 TI - A simple and safe operation method for brachiocephalic artery aneurysms. AB - A 64-year-old man was referred to our hospital due to dyspnea and fever. The chest computed tomogram revealed a 60-mm aneurysm of the brachiocephalic artery with mural thrombus. The aneurysm of the brachiocephalic artery and the right subclavian artery were exposed through only median sternotomy. Cardiopulmonary bypass with synchronized pulsatile perfusion was established with the ascending aorta and bi-caval cannulation. A mean arterial pressure was kept at between 60 and 70 mmHg with the unloaded beating heart. Mild hypothermia was induced (blood temperature 27 degrees C, nasopharyngeal temperature 32 degrees C). The brachiocephalic artery, right carotid artery, and right subclavian artery were clamped when nasopharyngeal temperature was 32 degrees C after decreasing blood temperature to 27 degrees C. After opening the aneurysm, the mural thrombus and calcified aneurysmal wall were removed. First, an ascending aorta to the right common carotid artery bypass was performed using a 16-8 mm Y- prosthetic graft with side-clamp forceps. After the anastomosis, the right side cerebral perfusion was restarted and the patient was rewarmed. Then the right subclavian artery was anastomosed in an end-to-end fashion. The duration of the right side cerebral circulatory arrest was 30 minutes. The patient left hospital seven days after the operation. PMID- 18384573 TI - Biatrial myxoma: a case report and review of the literature. AB - BACKGROUND: In surgical series, a majority of benign cardiac tumors are myxomas. Of these, only about 2.5% are biatrial. Only 10 cases have been reported in the last 10 years. We present here a successful case in a 51-year-old man. A brief review of the literature is presented to place this case in context. METHODS: The tumor was removed surgically via a midline sternotomy using cardiopulmonary bypass. Both left and right atrial extensions of the tumor mass were removed. The resection involved the entire septum, with a bovine patch used to reconstruct the atrial septum. RESULTS: Patient recovered uneventfully. We advised follow-up evaluation using transthoracic echocardiography annually. CONCLUSIONS: Biatrial myxoma is a very rare condition, with diagnostic challenges, but is amenable to modern surgical approaches. PMID- 18384574 TI - Life-threatening pulmonary embolism associated with a thrombus straddling a patent foramen ovale: report of a case. AB - BACKGROUND AND AIM: A biatrial thrombus straddling a patent foramen ovale (PFO) is rare. The optimal management is controversial. This report offers an additional report and brief review of the literature. CASE REPORT: A 72-year-old female presented with paroxysmal dyspnea. Transthoracic echocardiography showed distended right heart cavities, pulmonary artery hypertension, and a thrombus in the right atrium passing through the PFO into the left atrium. Urgent surgical embolectomy confirmed an 11.5 cm serpentine biatrial thrombus and allowed PFO closure and bilateral pulmonary embolectomy. Postoperative recovery was uneventful. CONCLUSIONS: The risk of systemic embolization during thrombolytic or heparin treatment for biatrial thrombus makes most authors recommend surgical or interventional thrombectomy and PFO closure. Given the limited number of cases, there is no evidence that any of the treatment strategies provide a better survival. PMID- 18384575 TI - Fractured inferior vena cava filter strut presenting as a penetrating foreign body in the right ventricle: report of a case. AB - Migration of a fractured strut of an inferior vena cava (IVC) filter to the heart is a rare complication. We report the case of a 40-year-old woman who had this complication eight months after infrarenal IVC filter placement. She presented with chest pain. The broken arm of the IVC filter had migrated to the heart and penetrated the free wall of the right ventricle. It was removed successfully by cardiac surgery without the aid of cardiopulmonary bypass. PMID- 18384576 TI - Endarterectomy and surgical angioplasty without cardiopulmonary bypass. AB - Surgical angioplasty for ostial stenosis of the major coronary arteries is a well established treatment modality. Management of discrete stenoses of distal coronary arteries in the presence of mild proximal disease is a challenging task. Percutaneous intervention or grafting beyond the diseased segment may be limited by size of the target vessel, whereas endarterectomy followed by graft placement may result in graft occlusion due to competitive native flow. Patch angioplasty with or without endarterectomy is an effective option in this setting, which can be performed without cardiopulmonary bypass in the appropriate group of patients. A patient with triple vessel coronary artery disease and a discrete lesion in the posterior descending artery underwent off-pump endarterectomy with vein patch angioplasty in combination with bypass grafting to left anterior descending and obtuse marginal arteries. Operative technique and intraoperative strategies are discussed. PMID- 18384577 TI - Implantable defibrillators and prevention of sudden death in hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HCM) is the most common cause of sudden cardiac death in young people, including trained athletes. The implantable cardioverter defibrillator (ICD), although initially designed as a treatment for older patients with coronary artery disease, has more recently proved to be a safe and effective therapeutic intervention in young patients with HCM, both for primary or secondary prevention of sudden death. The largest such report of >500 patients showed that the ICD intervened appropriately to abort ventricular tachycardia/fibrillation (VT/VF) in 20% of patients over an average follow-up period of only 3.7 years, at a rate of about 4% per year in those patients implanted prophylactically, and often with considerable delays of up to 10 years. Extensive experience with high-risk HCM patients showed that appropriate device discharges for VT/VF occur with similar frequency in patients with 1, 2, or > or = 3 noninvasive risk markers. Despite the extreme morphology characteristic of HCM, often with massive degrees of left ventricular (LV) hypertrophy and/or LV outflow tract obstruction, ICDs performed in a highly effective fashion, with failure to convert life-threatening arrhythmias extraordinarily rare. In conclusion, in a large high-risk HCM cohort, ICD interventions for life threatening ventricular tachyarrhythmias were frequent and highly effective in restoring normal rhythm. An important proportion of ICD discharges occurred in primary prevention patients with only one risk factor. Therefore, a single marker of high risk may represent sufficient evidence to justify the recommendation for a prophylactic ICD in selected patients with HCM. PMID- 18384578 TI - Health-related quality of life after bladder preservation therapy for muscle invasive bladder cancer. AB - OBJECTIVE: To assess health-related quality of life (QOL) of bladder cancer patients following bladder preservation therapy (BPT). METHODS: Eighty patients with muscle-invasive bladder cancer had been treated between January 1992 and July 2005 at our institutions with BPT consisting of transurethral resection, intra-arterial chemotherapy and radiotherapy. Among them, 48 were alive and free from recurrence at the time of survey and were asked to participate. A total of 168 patients who had been treated for superficial bladder cancer in the same period were used as a control group. Three questionnaires, namely the International Prostate Symptom Score (IPSS), the SF-36, and the Expanded Prostate Cancer Index Composite (EPIC) were used. RESULTS: Thirty-three patients in the BPT group (68.8%) and 128 patients in the control group (76.2%) answered the QOL survey. There was no significant difference in age, gender and other clinical factors among these two groups. No significant difference was found between the groups according to IPSS. The QOL score of BPT was lower than that of the control group in the SF-36, but there was no significant difference without body pain (P = 0.047). There was a tendency toward a diminished physical functioning (P = 0.053) and role-physical (P = 0.064) in BPT. The EPIC scores for urinary function, especially storage and voiding symptoms, and bowel function were significantly lower in the BPT group. At multivariable analysis, body pain and bowel function were associated with the type of treatment. CONCLUSION: Although some of the QOL outcome parameters after BPT were found to be lower than the control group, these differences were not significant. Overall, patients retaining their bladder had an acceptable health related QOL. PMID- 18384580 TI - Type 2 diabetes mellitus and cognitive decline in two large cohorts of community dwelling older adults. AB - OBJECTIVES: To relate diabetes mellitus (DM) status and duration to late-life cognitive impairment and decline in men and women. DESIGN: Prospective cohort. SETTING: Community. PARTICIPANTS: Five thousand nine hundred seven men in the Physicians' Health Study II and 6,326 women in the Women's Health Study (mean age 74.1 and 71.9, respectively, at baseline cognitive assessment); 553 men and 405 women had DM. MEASUREMENTS: Primary outcomes were general cognition (the Telephone Interview for Cognitive Status (TICS) and a global score averaging five tests) and verbal memory. All participants had second assessments approximately 2 years later; women had third assessments an average of 4 years later. RESULTS: In adjusted linear regression models, participants with DM had significantly lower baseline scores for all outcomes, and longer duration of DM was associated with lower scores (P-trends < .001). Men with DM had significantly greater 2-year cognitive decline than men without DM, and longer duration of DM was associated with worse decline (P-trends < or = .01). In repeated-measures analyses of response profiles, women with DM had significantly greater 4-year cognitive decline in all outcomes than women without DM. In women, as in men, there was generally greater cognitive decline with longer duration of DM (e.g., the adjusted mean difference in decline on the TICS associated with duration of > or = 5 years was -0.74 (95% confidence interval = -1.05 to -0.43) points (P-trend < .001). There were no significant sex-DM interactions. CONCLUSION: Type 2 DM and longer duration of DM are similarly related to cognitive impairment and decline in men and women. PMID- 18384581 TI - The effects of total hip arthroplasty on physical functioning in the older population. AB - OBJECTIVES: To estimate the effects of total hip arthroplasty (THA) on three levels of physical functioning in a representative national sample of older adults. DESIGN: Survey. SETTING: Participants were interviewed in their homes. PARTICIPANTS: Participants consisted of stratified random samples of Medicare beneficiaries interviewed between 1992 and 2003. METHODS: Data from the Medicare Current Beneficiary Survey from 1992 to 2003 and merged Medicare claims data identified participants who received (n=131) or did not receive (n=257) THA. Outcomes were three measures of physical functioning: Nagi items, instrumental activities of daily living, and activities of daily living. Baseline and follow up measures were obtained approximately 6 months apart. Logistic regression was used to predict the effects of THA on functioning, with a wide range of covariates controlled. RESULTS: Persons who received THA significantly improved in two of three levels of physical functioning; the no-treatment group experienced declines in physical functioning. CONCLUSION: Receipt of THA is associated with significant improvements in two levels of physical functioning. These benefits are broadly applicable in that a wide variety of covariates had minimal effects on THA receipt and outcome. PMID- 18384579 TI - Incident physical disability in people with lower extremity peripheral arterial disease: the role of cardiovascular disease. AB - OBJECTIVES: To evaluate the risk of incident physical disability and the decline in gait speed over a 6-year follow-up associated with a low ankle-arm index (AAI) in older adults. DESIGN: Observational cohort study. SETTING: Forsyth County, North Carolina; Sacramento County, California; Washington County, Maryland; and Allegheny County, Pennsylvania. PARTICIPANTS: Four thousand seven hundred five older adults, 58% women and 17.6% black, participating in the Cardiovascular Health Study. MEASUREMENTS: AAI was measured in 1992/93 (baseline). Self-reported mobility, activity of daily living (ADL), and instrumental activity of daily living (IADL) disability and gait speed were recorded at baseline and at 1-year intervals over 6 years of follow-up. Mobility disability was defined as any difficulty walking half a mile and ADL and IADL disability was defined as any difficulty with 11 specific ADL and IADL tasks. Individuals with mobility, ADL, or IADL disability at baseline were excluded from the respective incident disability analyses. RESULTS: Lower baseline AAI values were associated with increased risk of mobility disability and ADL/IADL disability. Clinical cardiovascular disease (CVD), diabetes mellitus, and interim CVD events partially explained these associations for mobility disability and clinical CVD and diabetes mellitus partially explained these associations for ADL and IADL disability. Individuals with an AAI less than 0.9 had on average a mean decrease in gait speed of 0.02 m/s per year, or a decline of 0.12 m/s over the 6-year follow-up. Prevalent CVD partly explained this decrease but interim CVD events did not further attenuate it. CONCLUSION: Low AAI serves as marker of future disability risk. Reduction of disability risk in patients with a low AAI should consider cardiovascular comorbidity and the prevention of additional disabling CVD events. PMID- 18384582 TI - Predictors of 6-month mortality in elderly patients with mild chronic obstructive pulmonary disease discharged from a medical ward after acute nonacidotic exacerbation. AB - OBJECTIVES: To identify clinical outcomes and variables associated with 6-month mortality in very elderly patients admitted for nonacidotic acute exacerbation of chronic obstructive pulmonary disease (AECOPD). DESIGN: Prospective cohort study. SETTING: General medicine acute care ward. PARTICIPANTS: Two hundred forty-four elderly patients with COPD (mean age+/-standard deviation 82+/-7, 55.7% female) admitted to the hospital because of non-acidotic AECOPD. MEASUREMENTS: Cognitive and mood status and physiological variables were measured. Self-reported comorbidities were assessed using the Charlson Comorbidity Index. In-hospital and long-term mortality and clinical outcomes were recorded. RESULTS: At admission, this elderly population with AECOPD had low cognitive performance (mean Mini Mental State Examination score 21+/-5), no presence of significant depressive symptoms (Geriatric Depression Scale score 4+/-3), good nutritional status (body mass index (BMI) 25.1+/-5.5), moderate comorbidity (Charlson Comorbidity Index 4.0+/-1.9), high functional disability (Barthel Index (BI) 52+/-34), and moderate severity of acute exacerbation (Acute Physiology and Chronic Health Evaluation (APACHE) II score 9.7+/-4.2). Two hundred twenty-five inpatients with AECOPD were successfully discharged, whereas 15 were transferred to the intensive care unit, and four died in the hospital. The 6-month cumulative mortality rate in discharged patients with AECOPD was 20%. Multivariate Cox analysis shows that lower BMI (beta=-0.16; 95% confidence interval (CI)=0.73-0.99), higher APACHE II score (beta=0,17; 95% CI=1.03-1.36), and lower BI at discharge (beta=-0.02; 95% CI=0.96-0.99) were independently associated with 6-month mortality. CONCLUSION: Malnutrition, severity of exacerbation and disability status could be identified as risk factors associated with 6-month mortality of elderly patients admitted for nonacidotic AECOPD. PMID- 18384583 TI - Carvedilol versus controlled-release metoprolol for elderly veterans with heart failure. AB - OBJECTIVES: To characterize prescribing of carvedilol and controlled-release (CR) metoprolol for veterans with heart failure who were predominantly elderly (aged > or = 65) and to compare the time to first hospitalization or death. DESIGN: Retrospective cohort. SETTING: Veterans Health Administration (VHA) nationwide healthcare system. SUBJECTS: Veterans enrolled in fee-for-service Medicare that had a VHA diagnosis of heart failure and a first prescription for carvedilol (n=17,429) or CR metoprolol (n=8,683) between October 1999 and September 2003. MEASUREMENTS: Prescribed daily doses and CR metoprolol-to-carvedilol hazard ratios (HRs) estimated using Cox regression within strata defined according to propensity to prescribe carvedilol. RESULTS: After several months, the majority of prescribed doses in both cohorts were less than 50% of target doses. The risk adjusted HR was 0.99 (95% confidence interval=0.96-1.03) for hospitalization or death and 0.91 (0.85-0.96) for death alone. CONCLUSION: Doses of beta-blockers being prescribed to predominantly elderly veterans with heart failure were much lower than proven doses. Whether efforts to increase doses can improve the effectiveness of beta-blocker therapy warrants further study. At the prescribed doses, CR metoprolol was associated with better survival than carvedilol, although unmeasured confounding variables might explain the observed difference in survival. PMID- 18384584 TI - Dual use of bladder anticholinergics and cholinesterase inhibitors: long-term functional and cognitive outcomes. AB - OBJECTIVES: To determine the cognitive and functional consequences of dual use of cholinesterase inhibitors (ChIs) and the bladder anticholinergics oxybutynin or tolterodine. DESIGN: Prospective cohort study. SETTING: Nursing homes (NHs) in the state of Indiana. PARTICIPANTS: Three thousand five hundred thirty-six Medicaid-eligible NH residents aged 65 and older taking a ChI between January 1, 2003, and December 31, 2004. Residents were excluded if they were taking an anticholinergic other than oxybutynin or tolterodine. MEASUREMENTS: Indiana Medicaid claims data were merged with data from the Minimum Data Set (MDS). Repeated-measures analyses were performed to assess the effects of dual therapy on change in cognitive function measured using the MDS Cognition Scale (MDS-COGS; scored 0-10) and change in activity of daily living (ADL) function using the seven ADL items in the MDS (scored 0-28). Potential covariates included age, sex, race, number of medications, and Charlson Comorbidity Index score. RESULTS: Three hundred seventy-six (10.6%) residents were prescribed oxybutynin or tolterodine concomitantly with a ChI. In residents in the top quartile of ADL function, ADL function declined an average of 1.08 points per quarter when not taking bladder anticholinergics (ChI alone), compared with 1.62 points per quarter when taking dual therapy, a 50% greater rate in quarterly decline in ADL function (P=.01). There was no excess decline attributable to dual therapy in MDS-COGS scores or in ADL function for residents who started out with lower functioning. CONCLUSION: In higher-functioning NH residents, dual use of ChIs and bladder anticholinergics may result in greater rates of functional decline than use of ChIs alone. The MDS COGS may not be sensitive enough to detect differences in cognition due to dual use. PMID- 18384585 TI - Association between unmet needs for medication support and all-cause hospitalization in community-dwelling disabled elderly people. AB - OBJECTIVES: To clarify the association between unmet medication management need and 3-year mortality and hospitalization for community-dwelling older people with various levels of disabilities. DESIGN: Prospective cohort study (the Nagoya Longitudinal Study for Frail Elderly). SETTING: Community-based. PARTICIPANTS: One thousand seven hundred seventy-two community-dwelling elderly subjects (611 men, 1,161 women). MEASUREMENTS: Data included the clients' demographic characteristics, a rating for basic and instrumental activities of daily living (ADLs), number of prescribed medications and physician-diagnosed chronic diseases, medication adherence, ability to manage medication, and presence or absence of medication assistance. Cox proportional hazard models and the Kaplan Meier method were used to assess the association between the medication management at baseline and mortality or hospitalization during a 3-year period. RESULTS: Of 1,772 participants, 681 reported no difficulty with self-medication management, and 1,091 experienced difficulty with self-medication. Of participants with difficulty with self-medication management, 929 had medication assistance, and 162 did not. During a 3-year follow up, 424 participants died, and 758 were admitted to hospitals. The baseline data demonstrated that participants not receiving medication assistance were younger and had better ADL status and fewer comorbidities. Multivariate Cox regression models adjusting for potential confounders showed that the lack of assistance in those who needed medication assistance was associated with hospitalization but not mortality during the study period. CONCLUSION: In community-dwelling disabled elderly people, lack of medication assistance in those needing medication support was associated with higher risk of hospitalization. PMID- 18384586 TI - The Confusion Assessment Method: a systematic review of current usage. AB - OBJECTIVES: To examine the psychometric properties, adaptations, translations, and applications of the Confusion Assessment Method (CAM), a widely used instrument and diagnostic algorithm for identification of delirium. DESIGN: Systematic literature review. SETTING: Not applicable. MEASUREMENTS: Electronic searches of PubMED, EMBASE, PsychINFO, CINAHL, Ageline, and Google Scholar, augmented by reviews of reference listings, were conducted to identify original English-language articles using the CAM from January 1, 1991, to December 31, 2006. Two reviewers independently abstracted key information from each article. PARTICIPANTS: Not applicable. RESULTS: Of 239 original articles, 10 (4%) were categorized as validation studies, 16 (7%) as adaptations, 12 (5%) as translations, and 222 (93%) as applications. Validation studies evaluated performance of the CAM against a reference standard. Results were combined across seven high-quality studies (N=1,071), demonstrating an overall sensitivity of 94% (95% confidence interval (CI)=91-97%) and specificity of 89% (95% CI=85-94%). The CAM has been adapted for use in the intensive care unit, emergency, and institutional settings and for scoring severity and subsyndromal delirium. The CAM has been translated into 10 languages where published articles are available. In application studies, CAM-rated delirium is most commonly used as a risk factor or outcome but also as an intervention or reference standard. CONCLUSION: The CAM has helped to improve identification of delirium in clinical and research settings. To optimize performance, the CAM should be scored based on observations made during formal cognitive testing, and training is recommended. Future action is needed to optimize use of the CAM and to improve the recognition and management of delirium. PMID- 18384587 TI - T-lymphocytes expressing CC chemokine receptor-5 are increased in frail older adults. AB - OBJECTIVES: To evaluate the frequencies of T-lymphocytes expressing CC chemokine receptor-5 (CCR5(+) T-cells) and their relationship with frailty in older adults. DESIGN: Case-control study with an age-, race-, and sex-matched design. SETTING: General Clinical Research Center. PARTICIPANTS: Community-dwelling adults aged 72 and older from Baltimore, Maryland. METHODS: Frailty was determined using five validated criteria: weakness, slow walking speed, fatigue, low physical activity, and weight loss. Those meeting three or more of these five criteria were defined as frail and those with none as nonfrail. Complete blood counts were performed to obtain peripheral lymphocyte counts using an automated (Coulter) counter. Peripheral blood was collected for surface immunofluorescent staining of CCR5 and other T-cell markers. RESULTS: Twenty-six frail and matched nonfrail participants (mean age+/-standard deviation 83.8+/-5.3, range 72-94) completed the study. Frail participants had higher CCR5(+), CCR5(+)CD8(+), and CCR5(+)CD45RO(-) T-cell counts than matched nonfrail controls (349+/-160/mm(3) vs 194+/-168/mm(3), P=.02; 208+/-98/mm(3) vs 105+/-62/mm(3), P=.02; and 189+/-149/mm(3) vs 52+/-36/mm(3), P=.01; respectively). Furthermore, there was a trend toward graded increase in these T-cell counts across the frailty scores in frail participants (e.g., CCR5(+)CD8(+) counts of 123+/-52/mm(3), 248+/-115/mm(3), and 360+/-215/mm(3) for those with frailty scores of 3, 4, and 5, respectively). CONCLUSION: These initial results suggest an expansion of the CCR5(+) T-cell subpopulation in frailty. They provide a basis for further characterization of CCR5(+) T-cells and their role in frailty, with potential therapeutic implications. PMID- 18384588 TI - Exploratory analysis of cerebral oxygen reserves during sleep onset in older and younger adults. AB - OBJECTIVES: To explore differences in cerebral oxygen reserves during sleep in old and young adults. DESIGN: Descriptive cross-sectional study. SETTING: General clinical research center. PARTICIPANTS: Nine old (aged 65-84) and 10 young (aged 21-39) adults. MEASUREMENTS: Subjects were monitored during the first nightly sleep cycle using standard polysomnography, including measures of arterial oxyhemoglobin saturation (SaO(2)). Changes in regional cerebral oxyhemoglobin saturation (rcSO(2)) were used to estimate cerebral oxygen reserves. General linear models were used to test group differences in the change in SaO(2) and rcSO(2) during sleep. RESULTS: Older subjects had lower SaO(2) than young subjects before sleep (baseline) (F((1,18))=5.1, P=.04) and during sleep (F((1,18))=10.7, P=.01). During sleep, half of the older subjects and none of the younger ones had SaO(2) values below 95%. In addition, the older subjects had more periods of oxygen desaturation (drops in SaO(2) > or = 4%) (chi-square=24.3, P=.01) and lower SaO(2) levels during desaturation (F((1,18))=11.1, P<.01). Although baseline values were similar, rcSO(2) decreased during sleep 2.1% in older subjects (F((1,8))=3.8, P=.05) but increased 2.1% during sleep in younger subjects (F((1,9))=4.6, P=.04). When the older subjects awakened from sleep, rcSO(2), but not SaO(2), returned to baseline; both returned to baseline in younger subjects. CONCLUSION: This exploratory analysis generated the hypothesis that lower SaO(2), combined with declines in regional blood flow, contributes to decline in cerebral oxygen reserves during sleep in older subjects. Further study will assess the effects of factors (e.g., medical conditions, subclinical disorders, and sleep architecture) that might account for these differences. PMID- 18384589 TI - Cognitive, functional, and quality-of-life outcomes of patients aged 80 and older who survived at least 1 year after planned or unplanned surgery or medical intensive care treatment. AB - OBJECTIVES: To investigate long-term cognitive, functional, and quality-of-life outcomes in very elderly survivors at least 1 year after planned or unplanned surgery or medical intensive care treatment. DESIGN: Retrospective cohort study. SETTING: General, 1,024-bed, tertiary university teaching hospital in The Netherlands. PARTICIPANTS: Two hundred four survivors of a cohort of 578 patients admitted to the medical-surgical intensive care unit (ICU) between January 1997 and December 2002 and alive in December 2003. The majority of survivors underwent elective surgery. MEASUREMENTS: From December 2003 until February 2004, data were collected from 190 patients and 169 relatives. The measures were: Informant Questionnaire on Cognitive Decline short form (IQCODE-SF) (cognition), modified Katz index of activities of daily living (ADLs) (functional status), and EuroQol (EQ-5D) (health-related quality of life). The patients themselves completed the modified Katz ADL index and EQ-5D forms; their caregivers completed the ADL caregiver version and IQCODE-SF. RESULTS: The mean age at admission+/-standard deviation was 81.7+/-2.4, and the median time after discharge was 3.7 years (range 1-5.9 years). Of the ICU patients who had planned surgery, 57% survived, compared with 11% of the unplanned surgical admissions and 10% of the medical patients. Three-quarters (74.3%) of the patients who lived at home before ICU admission remained at home at follow-up. Eighty-three percent had no severe cognitive impairment, and 76% had no severe physical limitations (33% had moderate, 40% had mild, and 3% had no limitations). The perceived quality of life was similar to that of an age-matched general population. CONCLUSION: Long-term survivors of ICU treatment received at the age of 80 and older showed fair-to good cognitive and physical functioning and quality of life, although few patients who underwent unplanned surgery or who were admitted to the ICU for medical reasons survived. PMID- 18384591 TI - Cartilaginous choristoma of the tongue. AB - Choristomas are lesions composed of normal cells or tissues occurring in an abnormal location. Cartilaginous choristomas of the oral mucosa are rare and occur preferentially on the tongue and less often in sites such as the soft palate and gingiva. Oral lesions are generally covered by integral mucosa and can occur at any age. The present study describes a case of a 73-year-old female presenting with an asymptomatic cartilaginous choristoma on the ventral surface of the tongue which had developed over a period of 3 years. The clinical presentation and management of the case are discussed and the literature is reviewed. This is the 28th reported case of a cartilaginous choristoma of the tongue and the third with a ventral localisation. PMID- 18384590 TI - Determining care management activities associated with mastery and relationship strain for dementia caregivers. AB - OBJECTIVES: To identify specific care management activities within a dementia care management intervention that are associated with 18-month change in caregiver mastery and relationship strain. DESIGN: Exploratory analysis, using secondary data (care management processes and caregiver outcomes) from the intervention arm of a clinic-level randomized, controlled trial of a dementia care management quality improvement program. SETTING: Nine primary care clinics in three managed care and fee-for-service southern California healthcare organizations. PARTICIPANTS: Two hundred thirty-eight pairs: individuals with dementia and their informal, nonprofessional caregivers. MEASUREMENTS: Care management activity types extracted from an electronic database were used as predictors of caregiver mastery and relationship strain, which were measured through mailed surveys. Multivariable linear regression models were used to predict caregiver mastery and relationship strain. RESULTS: For each care manager home environment assessment, caregiver mastery increased 4 points (range 0-100, mean+/-standard deviation 57.1+/-26.6, 95% confidence interval (CI)=2.4-5.7; P=.001) between baseline and 18 months. For every action linking caregivers to community agencies for nonspecific needs, caregiver mastery decreased 6.2 points (95% CI=-8.5 to -3.9; P<.001). No other care management activities were significantly associated with this outcome, and no specific activities were associated with a change in caregiver relationship strain. CONCLUSION: Home assessments for specific needs of caregivers and persons with dementia are associated with improvements in caregivers' sense of mastery. Future work is needed to determine whether this increase is sustained over time and decreases the need for institutionalization. PMID- 18384592 TI - Ground water recharge and flow characterization using multiple isotopes. AB - Stable isotopes of delta(18)O, delta(2)H, and (13)C, radiogenic isotopes of (14)C and (3)H, and ground water chemical compositions were used to distinguish ground water, recharge areas, and possible recharge processes in an arid zone, fault bounded alluvial aquifer. Recharge mainly occurs through exposed stream channel beds as opposed to subsurface inflow along mountain fronts. This recharge distribution pattern may also occur in other fault-bounded aquifers, with important implications for conceptualization of ground water flow systems, development of ground water models, and ground water resource management. Ground water along the mountain front near the basin margins contains low delta(18)O, (14)C (percent modern carbon [pmC]), and (3)H (tritium units [TU]), suggesting older recharge. In addition, water levels lie at greater depths, and basin bounding faults that locally act as a flow barrier may further reduce subsurface inflow into the aquifer along the mountain front. Chemical differences in ground water composition, attributed to varying aquifer mineralogy and recharge processes, further discriminate the basin-margin and the basin-center water. Direct recharge through the indurated sandstones and mudstones in the basin center is minimal. Modern recharge in the aquifer is mainly through the broad, exposed stream channel beds containing coarse sand and gravel where ground water contains higher delta(18)O, (14)C (pmC), and (3)H (TU). Spatial differences in delta(18)O, (14)C (pmC), and (3)H (TU) and occurrences of extensive mudstones in the basin center suggest sluggish ground water movement, including local compartmentalization of the flow system. PMID- 18384593 TI - Upgrading aquifer test analysis. PMID- 18384594 TI - Long-term reactive solute transport in an aquitard using a centrifuge model. AB - Characterizing and predicting reactive solute transport in low hydraulic conductivity (K) clay-rich media is challenging because the very long transport time for solutes renders conventional column tests impractical. In this study, a centrifugation technique was developed to assess the transport of a simple aqueous solution (NaCl) by accelerating flow by centrifugal force through low K (1.1 x 10(-11) m/s) core samples. Duplicate cores (52-mm length x 33-mm diameter) were centrifuged at 330 xg for 90 d to model the migration of saline pore water (0.5 M NaCl) under in situ conditions through an approximately 17-m-thick clay prototype over approximately 24,000 years. A PHREEQC one-dimensional reactive solute transport code simulated effluent breakthrough of the NaCl during centrifugation, with best-fit cation exchange coefficients similar to batch tests. The calibrated code was used to predict solute profile development over the long term in the prototype or simulated field-scale conditions. Chromatographic separation of solutes due to ion exchange was evident over several meters in the simulated prototype and the field profile. The applicability of centrifugation methods to predict transport of more complex suites of reactive solutes over the long term is yet to be verified. PMID- 18384595 TI - The value of subsidence data in ground water model calibration. AB - The accurate estimation of aquifer parameters such as transmissivity and specific storage is often an important objective during a ground water modeling investigation or aquifer resource evaluation. Parameter estimation is often accomplished with changes in hydraulic head data as the key and most abundant type of observation. The availability and accessibility of global positioning system and interferometric synthetic aperture radar data in heavily pumped alluvial basins can provide important subsidence observations that can greatly aid parameter estimation. The aim of this investigation is to evaluate the value of spatial and temporal subsidence data for automatically estimating parameters with and without observation error using UCODE-2005 and MODFLOW-2000. A synthetic conceptual model (24 separate cases) containing seven transmissivity zones and three zones each for elastic and inelastic skeletal specific storage was used to simulate subsidence and drawdown in an aquifer with variably thick interbeds with delayed drainage. Five pumping wells of variable rates were used to stress the system for up to 15 years. Calibration results indicate that (1) the inverse of the square of the observation values is a reasonable way to weight the observations, (2) spatially abundant subsidence data typically produce superior parameter estimates under constant pumping even with observation error, (3) only a small number of subsidence observations are required to achieve accurate parameter estimates, and (4) for seasonal pumping, accurate parameter estimates for elastic skeletal specific storage values are largely dependent on the quantity of temporal observational data and less on the quantity of available spatial data. PMID- 18384596 TI - Reducing capture zone uncertainty with a systematic sensitivity analysis. AB - The U.S. Environmental Protection Agency has established several methods to delineate wellhead protection areas (WHPAs) around community wells in order to protect them from surface contamination sources. Delineating a WHPA often requires defining the capture zone for a well. Generally, analytical models or arbitrary setback zones have been used to define the capture zone in areas where little is known about the distribution of hydraulic head, hydraulic conductivity, or recharge. Numerical modeling, however, even in areas of sparse data, offers distinct advantages over the more simplified analytical models or arbitrary setback zones. The systematic approach discussed here calibrates a numerical flow model to regional topography and then applies a matrix of plausible recharge to hydraulic conductivity ratios (R/K) to investigate the impact on the size and shape of the capture zone. This approach does not attempt to determine the uncertainty of the model but instead yields several possible capture zones, the composite of which is likely to contain the actual capture zone. A WHPA based on this composite capture zone will protect ground water resources better than one based on any individual capture zone. An application of the method to three communities illustrates development of the R/K matrix and demonstrates that the method is particularly well suited for determining capture zones in alluvial aquifers. PMID- 18384598 TI - Ground water budget analysis and cross-formational leakage in an arid basin. AB - Ground water budget analysis in arid basins is substantially aided by integrated use of numerical models and environmental isotopes. Spatial variability of recharge, storage of water of both modern and pluvial age, and complex three dimensional flow processes in these basins provide challenges to the development of a good conceptual model. Ground water age dating and mixing analysis with isotopic tracers complement standard hydrogeologic data that are collected and processed as an initial step in the development and calibration of a numerical model. Environmental isotopes can confirm or refute a priori assumptions of ground water flow, such as the general assumption that natural recharge occurs primarily along mountains and mountain fronts. Isotopes also serve as powerful tools during postaudits of numerical models. Ground water models provide a means of developing ground water budgets for entire model domains or for smaller regions within the model domain. These ground water budgets can be used to evaluate the impacts of pumping and estimate the magnitude of capture in the form of induced recharge from streams, as well as quantify storage changes within the system. The coupled analyses of ground water budget analysis and isotope sampling and analysis provide a means to confirm, refute, or modify conceptual models of ground water flow. PMID- 18384599 TI - Modeling axisymmetric flow and transport. AB - Unmodified versions of common computer programs such as MODFLOW, MT3DMS, and SEAWAT that use Cartesian geometry can accurately simulate axially symmetric ground water flow and solute transport. Axisymmetric flow and transport are simulated by adjusting several input parameters to account for the increase in flow area with radial distance from the injection or extraction well. Logarithmic weighting of interblock transmissivity, a standard option in MODFLOW, can be used for axisymmetric models to represent the linear change in hydraulic conductance within a single finite-difference cell. Results from three test problems (ground water extraction, an aquifer push-pull test, and upconing of saline water into an extraction well) show good agreement with analytical solutions or with results from other numerical models designed specifically to simulate the axisymmetric geometry. Axisymmetric models are not commonly used but can offer an efficient alternative to full three-dimensional models, provided the assumption of axial symmetry can be justified. For the upconing problem, the axisymmetric model was more than 1000 times faster than an equivalent three-dimensional model. Computational gains with the axisymmetric models may be useful for quickly determining appropriate levels of grid resolution for three-dimensional models and for estimating aquifer parameters from field tests. PMID- 18384600 TI - Foreword: ground water in arid zones. PMID- 18384601 TI - Perioral granulomatous reaction to Dermalive. PMID- 18384602 TI - Successful treatment of nail bed hypoplasia after nail loss by acrylic nail. PMID- 18384603 TI - Dermoscopy identifies histopathologically indiscernible malignant lesion of atypical melanosis of the foot, an early lesion of acral lentiginous melanoma in situ. PMID- 18384604 TI - Nail melanoma in children: differential diagnosis and management. PMID- 18384605 TI - Excision of a peristomal basal cell carcinoma using Mohs micrographic surgery. PMID- 18384606 TI - The painful leg. PMID- 18384607 TI - Basal cell carcinoma masquerading as the chronic ectropion of lamellar ichthyosis. PMID- 18384608 TI - Fordyce spots of the lip responding to electrodesiccation and curettage. PMID- 18384609 TI - Keratoacanthoma-like reaction after a hyaluronic acid and acrylic hydrogel cosmetic filler. PMID- 18384610 TI - Aggressive shaving after combined liposuction and curettage for axillary hyperhidrosis leads to more complications without further benefit. PMID- 18384611 TI - Pigmentation due to stasis dermatitis treated successfully with a noncoherent intense pulsed light source. PMID- 18384612 TI - Blue foot: an unusual complication of tattoo and successful treatment with a QS Nd:YAG laser. PMID- 18384613 TI - Nonsurgical rhinoplasty: an evaluation of injectable calcium hydroxylapatite filler for nasal contouring. PMID- 18384614 TI - Cerclage technique for repairing large circular defects of the trunk: two-staged excision of a plexiform neurofibroma. PMID- 18384615 TI - Primary cutis verticis gyrata and scalp reduction in one stage with multiple pinwheel flaps (revisited). PMID- 18384616 TI - Repair of a defect on the ala. PMID- 18384617 TI - Transplantation of autologous keratinocyte suspension in fibrin matrix to chronic venous leg ulcers: improved long-term healing after removal of the fibrin carrier. AB - BACKGROUND: The transplantation of keratinocytes suspended in fibrin carrier represents a candidate regimen for chronic ulcer treatment in an outpatient setting. We evaluated the integration and survival of autologous individualized keratinocytes applied within fibrin matrix onto chronic venous leg ulcers in vivo. Parallel in vitro culture was used to validate keratinocyte survival and apoptosis in fibrin compared to collagen matrix carrier. METHODS: Seven patients with chronic venous leg ulcers were transplanted with autologous keratinocytes suspended in fibrin sealant after isolation and expansion from full-skin biopsy. The fibrin carrier was removed in three patients after 7 days, whereas four patients served as control with fibrin remaining. In parallel in vitro cultures, primary keratinocyte movement in fibrin as well as viability in three-dimensional (3D) fibrin versus collagen lattices was examined. RESULTS: Complete ulcer healing was observed in four of seven ulcers after a mean duration of 14.5 weeks. If the fibrin layer was removed, complete wound healing occurred in three of three patients, compared to one of four in the control group. In vitro, keratinocytes formed a monolayer underneath but remained isolated and nonmobile within the fibrin matrix, suggesting reepithelialization along the lower fibrin interphase. Keratinocyte culture in 3D fibrin at clinically used concentration (90 mg/mL) caused high levels of apoptosis, similar to 3D collagen, which was prevented by diluting fibrin concentration to 3 mg/mL. CONCLUSIONS: Transplantation of autologous keratinocytes suspended in fibrin is efficient in the treatment of chronic venous leg ulcers. Due to an antimigratory and survival compromising effect, the presently used fibrin carrier should be removed after a few days of transplantation. PMID- 18384618 TI - Radiowave surgery versus CO laser for upper blepharoplasty incision: which modality produces the most aesthetic incision? AB - INTRODUCTION: For years, traditional upper blepharoplasty incisions have been made with scalpel and or scissors. Although effective, increased intraoperative bleeding can be problematic. Bleeding slows the surgery; obscures the surgical field; and can lead to increased swelling, bruising, and pain. Bloodless modalities for upper blepharoplasty include radiowave surgery, electrocautery, and CO(2) laser technology. These modalities provide a virtually dry and bloodless surgical field, which translates into faster surgery as well as decreased postoperative bleeding, bruising, and pain. PURPOSE: The purpose of this study was to determine which modality (CO(2) laser vs. 4.0-MHz radiowave surgery) produced the most aesthetic postoperative upper blepharoplasty scar at 1 year in a consecutive cohort of patients operated by the same surgeon. A search of the literature does not show a similar study in Caucasian patients. The mechanics and physics of CO(2) laser and 4.0-MHz radiowave surgery are also discussed. MATERIALS AND METHODS: Thirty consecutive patients underwent upper eyelid blepharoplasty for cosmetic purposes performed by the author. In all patients one upper eyelid was treated (skin, muscle, and fat) with a 4.0-MHz radiowave surgery unit (Surgitron, Ellman International) set at 12 W on the cut/coag mode, and the contralateral side was treated with an ultrapulse CO(2) laser (Encore, Lumenis Inc.) using a 0.8-mm handpiece on the continuous wave setting at 8 W. Six blepharoplasty-experienced, blinded observers consisting of doctors from five different cosmetic specialties evaluated standardized digital images of each patient taken 1 year after surgery. The photographs were randomized, and the blinded examiners were asked to choose the side that had the most esthetic postoperative incision. RESULTS: Of the 30 consecutive patients, 23 completed the required 1-year follow-up. Surgeons evaluating these patients scored the radiowave surgery side to look best in 37% of the cases (43/115), the CO(2) laser side was scored to look better in 37% (42/115), and both sides to be to be equally aesthetic in 26% of the cases. CONCLUSION: Both 4.0-MHz radiowave surgery and CO(2) laser incision produce simultaneous incision and coagulation. Both modalities restrict blood loss in the average four lid blepharoplasty surgery to less than 1 cm(3) of blood and decrease operative time. In this study, qualified blinded surgeons judging 12-month postblepharoplasty photos of incisional scars were unable to differentiate a statistically significant difference between the two modalities and the aesthetic quality of the scars. This represents the first study of its type in the literature utilizing Caucasian patients. PMID- 18384619 TI - Rejuvenating influence of a stabilized hyaluronic acid-based gel of nonanimal origin on facial skin aging. AB - BACKGROUND: Skin changes associated with aging, such as loss of elasticity and turgor, can have a negative psychosocial impact. Current treatment options aimed at restoring elasticity are often ineffective or hampered by a poor risk/benefit ratio. OBJECTIVE: Clinical observations and biophysical measurements were used to assess the effect of middermal placement of stabilized hyaluronic acid-based gel of nonanimal origin (NASHA, Restylane, Vital, Q-Med) on the appearance and elasticity of facial skin. METHODS AND MATERIALS: In this pilot study, 19 female patients underwent a series of three treatment sessions, spaced 4 weeks apart, with NASHA injected into the lower cheeks. Elasticity, skin surface roughness, dermal thickness, and density were evaluated at each treatment session and at 4 and 12 weeks after the last treatment session. Patient satisfaction was assessed, and photographs were taken at each visit. RESULTS: During the course of the study, skin elasticity and surface roughness improved significantly. Patient feedback was extremely positive. CONCLUSION: Micropuncture placement of NASHA can exert a rejuvenating effect on facial skin. PMID- 18384620 TI - Successful treatment of penile Kaposi's sarcoma with electrochemotherapy. PMID- 18384621 TI - Bilateral periventricular nodular heterotopia and lissencephaly in an infant with unbalanced t(12;17)(q24.31; p13.3) translocation. AB - Periventricular nodular heterotopia and Miller-Dieker syndrome are two different disorders of brain development. Miller-Dieker syndrome exhibits classical lissencephaly and is related to defects in the lissencephaly gene (LIS1). Periventricular nodular heterotopia is characterized by aggregates of grey matter adjacent to the lateral ventricle and is mainly linked to mutations in the Filamin A (FLNA) gene. We describe a male infant presenting with facial dysmorphisms resembling those of Miller-Dieker syndrome, neuromotor delay, and drug - resistant infantile spasms. Magnetic resonance imaging of the brain showed periventricular nodular heterotopia overlaid by classical lissencephaly with complete agyria. Cytogenetic and molecular investigations detected a maternally inherited unbalanced translocation involving chromosome arms 17p and 12q. This resulted in partial monosomy of 17p13.3-->pter and partial trisomy of 12q24.3- >qter. No mutation was found in the FLNA gene. The patient died at the age of 22 months from respiratory insufficiency during an infection of the lower respiratory tract. Our observation extends the list of the overlying cortical malformations associated with periventricular nodular heterotopia. It remains to be established whether this peculiar neuronal migration disorder represents a phenotype totally linked to 17q13.3 deletion or results from a combination of gene defects at 17q13.3 and 12q24.3. PMID- 18384622 TI - Are baroreflex events detected by invasive and non-invasive techniques coincident? AB - Cardiac baroreceptor sensitivity, a prognostic indicator for a range of diseases, such as myocardial infarction and stroke, may be estimated from spontaneous fluctuations of arterial blood pressure (BP) and heart rate using sequence analysis. We tested the hypothesis that BP values recorded with the non-invasive Finapres device do not always produce sequences coincident with sequences detected from central BP measurements. Finapres recordings of resting BP in the finger, ascending aorta (Millar catheter-tip transducer) and ECG were obtained from 34 patients undergoing coronary angioplasty, including 24 patients treated with betablockers. Coincidence of baroreflex sensitivity (BRS) sequences was expressed by the sensitivity of the Finapres to detect a simultaneously occurring sequence in aortic pressure. The influence of different criteria to detect and accept sequences from beat-to-beat values of systolic BP (SBP) and cardiac interval (RRi) on the Finapres sensitivity was also assessed. The Finapres was able to detect 70.7% of all three beat intra-arterial sequences when the selection criteria was based on the correlation coefficient between SBP and RRi (>0.85), but decreased to 27.5% when the P-value of the linear regression was limited to 0.05. Changing the thresholds for minimum changes in SBP and RRi also had significant effects on sensitivity, as well as in the corresponding values of BRS. Significant differences in BRS were obtained between invasive and non invasive estimates, but there was no difference between non-invasive estimates calculated from coincident and non-coincident sequences. Non-invasive, compared with intra-arterial estimates of BRS by sequence analysis are not influenced by coincidence of sequences if acceptance of sequences is based on the correlation coefficient criteria (>0.85). PMID- 18384623 TI - Integrin scintimammography using a dedicated breast imaging, solid-state gamma camera and (99m)Tc-labelled NC100692. AB - OBJECTIVE: Integrin scintimammography with NC100692 and a dedicated gamma-camera, LumaGEM, based on semi-conductor technology, was performed to investigate the detection ability of this combination in breast cancer. METHODS: Eight patients with a high suspicion of breast cancer were administered 600-750 MBq (99m)Tc labelled NC100629. Two acquisitions using a cranio-caudal and a lateral view were used, with moderate compression of the breast. RESULTS: LumaGEM scintigraphy revealed 9 of 11 malignancies, sized 6-20 mm. Two lesions in patients with multicentric disease were not diagnosed, one of which measured only 2.5 mm. In one patient, the procedure was inconclusive, due to major breast hypertrophy. CONCLUSION: The combined use of NC100629 and a dedicated gamma-camera for breast imaging was highly effective in diagnosing breast cancer. PMID- 18384624 TI - Impaired nitroglycerin biotransformation in patients with chronic heart failure. AB - OBJECTIVE: Patients with chronic heart failure (CHF) often require higher doses of nitroglycerin (glyceryl trinitrate, GTN) than patients with normal cardiac function to achieve a given haemodynamic goal. Two pathways leading to biotransformation of GTN have been characterized; a high-affinity pathway operative in nanomolar concentration ranges yielding predominantly 1,2-glyceryl dinitrate (1,2-GDN), and a low-affinity pathway operative at higher, micromolar concentrations of GTN associated with a greater proportion of 1,3-GDN formation. We tested the hypothesis that, at a given GTN-induced blood pressure reduction, the CHF group would present with: (i) higher concentrations of GTN; and (ii) decreased ratios of 1,2-GDN/GTN and 1,2-GDN/1,3-GDN compared with healthy subjects (HS). METHODS: Twelve patients with CHF (left ventricular ejection fraction 20 +/- 5%, NYHA III) and nine HS were investigated during a right cardiac catheterization. GTN was titrated intravenously until mean arterial blood pressure (MAP) was reduced by 15%. RESULTS: At arterial GTN concentrations of 27.2 [10.0-57.8] nmol l(-1) in CHF and 2.8 [2.5-3.5] nmol l(-1) in HS [median (quartile range), P<0.05 between groups], MAP and mean capillary wedge pressures were reduced similarly in both groups (approx. 15% and 65%, respectively, P = NS between groups). The ratios of 1,2-GDN/GTN and 1,2-GDN/1,3-GDN were lower in CHF (0.86 [0.28-1.58] and 5.8 [5.6-6.3]) compared with HS [1.91 (1.54-2.23) and 7.6 (7.2-10.2), P<0.05], with a negative correlation between the 1,2-GDN/1,3-GDN ratio and the arterial GTN concentrations in the CHF patients (R = -0.8, P<0.05). CONCLUSION: Patients with CHF have attenuated GTN responsiveness and decreased relative formation of 1,2-GDN in comparison with HS, indicating an altered biotransformation of GTN. PMID- 18384625 TI - Intravesically administered antisense oligonucleotides targeting heat-shock protein-27 inhibit the growth of non-muscle-invasive bladder cancer. AB - OBJECTIVE: To evaluate the inhibitory effects of a second-generation antisense oligonucleotide (ASO) targeting the cytoprotective chaperone heat-shock protein (HSP)-27 (OGX-427, OncoGeneX Technologies, Vancouver, Canada) on human bladder cancer growth both in vitro and in vivo as an intravesical agent in an orthotopic murine model. MATERIALS AND METHODS: Human KU-7 bladder tumour cells were treated with OGX-427 or a mismatch (MM) control oligodeoxynucleotide (ODN) in vitro and were assessed for HSP27 expression, proliferation and apoptosis. KU-7-luc cells that stably express luciferase were inoculated in female nude mice by intravesical instillation and tumour size was measured using bioluminescence imaging. Mice with established KU-7-luc tumours were administered uncomplexed 'naked' OGX-427 or MM ODN as well as controlled-release microparticulate chitosan/oligonucleotide formulations intravesically. Tumour growth was monitored over time and tumours were analysed after death using immunohistochemistry and Western blotting. RESULTS: In vitro, OGX-427 significantly decreased HSP27 protein levels and cellular viability. While naked OGX-427 showed only a trend in tumour suppression compared with MM ODN, OGX-427 complexed with chitosan significantly inhibited orthotopic tumour growth. The chitosan preparation induced some haematuria compared to naked ASO, but this formulation had superior tissue uptake of oligonucleotides and suppressed HSP27 tissue levels by 75%. CONCLUSION: Intravesical OGX-427 instillation therapy showed promising antitumour activity and minimal toxicity in an orthotopic mouse model of high-grade bladder cancer. These findings provide pre-clinical proof-of-principle for the use of ASO as intravesical agents for non-muscle-invasive bladder cancer, and warrant further evaluation of efficacy and safety in early-phase clinical trials. PMID- 18384626 TI - The relationship between the local and systemic inflammatory responses and survival in patients undergoing resection for localized renal cancer. AB - OBJECTIVE: To examine the relationship between the systemic inflammatory response (C-reactive protein, CRP), tumour interleukin-6 receptor and cyclooxygenase (COX) 2 expression, tumour T-lymphocytic (CD4+, CD8+) infiltration and cancer survival in patients undergoing resection for renal cell carcinoma (RCC), as both the local and systemic inflammatory responses appear to predict the outcome in these patients. PATIENTS AND METHODS: The study included 60 patients undergoing nephrectomy for localized RCC. Pre-operative circulating CRP levels were measured and tumour interleukin-6 receptor and COX-2 expression, tumour CD4+ and CD8+ T lymphocytes were assessed using immunohistochemical analysis. RESULTS: The median follow-up was 78 months, with 14 patients relapsing from their disease and nine cancer-specific deaths. On univariate and multivariate survival analysis, tumour stage and grade and CRP levels were identified as significant factors associated with relapse-free and cancer-specific survival. There was a significant direct relationship between Fuhrman grade and CD4+ T-lymphocytic infiltrate (P < 0.05). An increase in tumour expression of interleukin-6 receptor was weakly associated with an increase in tumour CD8+ T-lymphocytic infiltration (P = 0.057). An increase in tumour CD4+ T-lymphocytic infiltration was associated with an increase in CD8+ T-lymphocytic infiltration (P < 0.01). CONCLUSIONS: The present results suggest that tumour-based factors such as interleukin-6 receptor and COX 2 expression or T-lymphocytic subset infiltration are subordinate to systemic factors such as CRP level in determining survival in patients with localized RCC. PMID- 18384627 TI - Enucleation of renal cell carcinoma with ablation of the tumour base. AB - OBJECTIVE: To retrospectively assess the effectiveness of cancer control with enucleation of renal cell carcinoma (RCC), which is surgically expedient, allows preservation of maximal renal parenchyma, and makes intraoperative renal ischaemia unnecessary, by two surgeons routinely enucleating renal tumours and ablating the tumour bed with argon beam and the Nd-YAG laser. PATIENTS AND METHODS: Between 1996 and 2006 at our institution, 97 patients had RCC enucleated, with ablation of the tumour base. Patients with lesions other than RCC and those with von Hippel-Lindau disease or Birt-Hogg-Dube syndrome were excluded from the study. The mean follow-up was 24.9 months. Patients were evaluated for RCC recurrence with cross-sectional imaging at least every 6 months for the first 2 years and then annually thereafter. RESULTS: The mean (median, range) tumour size was 2.8 (2.5, 0.8-7.0) cm. Of the 97 patients only one had disease progression after a mean follow-up of 24.9 months. This patient presented with a solitary grade 2 clear cell RCC and had a local recurrence 30 months after original surgery. CONCLUSIONS: The present series and other available clinical data suggest that enucleation with cavity ablation is an oncologically sound approach that is simple, versatile and obviates the need for renal ischaemia. PMID- 18384628 TI - Molecular and histological markers in urothelial carcinomas of the upper urinary tract. AB - Urothelial cell carcinomas (UCCs) are one of the most common types of malignancies. Recently, different mechanisms of carcinogenesis, as well as discrepancies in the natural history of urothelial cancers of the bladder and of the upper urinary tract (UUT), have been identified. As a result several teams have focused on specific markers in UUT-UCCs, a very rare type of cancer. This review gives a brief overview on the current markers of interest. Microsatellite instabilities (MSI) are independent molecular makers for prognosis. In addition, MSI can help detect a germline mutation and therefore allows for the detection of possible hereditary cancers. The loss of proteins of the mismatch repair system can also facilitate the detection of a germline mutation but should be followed by DNA sequencing. Epithelial cadherin has been shown to be an independent marker of prognosis, as well as hypoxia-inducible factor-1alpha (HIF-1alpha) and telomerase RNA component. Furthermore HIF-1alpha is significantly associated with the grade and pattern of growth and the telomerase RNA component could possibly also be used in diagnosis. The active form of the L-type amino acid transporter 1 (LAT1) was a significant prognostic marker in univariate analysis only. There are contrasting studies on the significances of p27 and Ki-67 as prognostic markers in UUT-UCCs. MET is a factor that correlates with vascular invasion of invasive cancer and bcl-2 oncoprotein correlates with stage. The ongoing identification of these markers might help to find specific treatments tailored to the molecular pattern of each tumour. Therefore a subgroup of patients with a higher risk of recurrence could be identified as well as patients that could benefit from minimal invasive surgery. PMID- 18384629 TI - DNA polymorphisms in exon 1 and promoter of the CDH1 gene and relevant risk of transitional cell carcinoma of the urinary bladder. AB - OBJECTIVE: To investigate the association between DNA polymorphisms, including single-nucleotide polymorphisms (SNPs) and insertion/deletion polymorphisms, in exon 1 and promoter of the CDH1 gene, and the risk of transitional cell carcinoma (TCC) of the urinary bladder (TCCB). PATIENTS, SUBJECTS AND METHODS: This was a hospital-based case-control study of 180 patients with TCCB and 110 normal controls. Genomic DNA was extracted from blood samples of all participants and genotypes determined using polymerase chain reaction and DNA sequencing techniques. Haplotypes were analysed using appropriate software. RESULTS: SNPs were detected at -160A/C, -73A/C and 178C/T; an inserted oligonucleotide of 5' CCGTGCCCCAGCC-3' was identified at 234 bp. The -160A allele frequency in the case group was 0.67, statistically higher than in the control group (0.42; P < 0.001), and higher in invasive carcinoma (0.77) than in superficial carcinoma (0.60). For -73C/A and 178C/T SNPs there was no difference among genotypes. The 234 repeat oligonucleotide insertion (2I) frequencies in cases was 0.27, statistically higher than in the control group (0.17; P = 0.01). The most common haplotype in controls was C-A-T-I (28%), the frequency of which was higher than in the TCCB group (6%). The A-A-T-2I was the only variation distribution carrying the -160A allele and was at a statistically higher frequency in the TCCB group (37%, the most common haplotype in cases) than in the control group. CONCLUSION: The -160A, 234 2I allele and haplotype A-A-T-2I were risk factors of TCCB. Haplotype C-A-T-I might act as a protective factor for TCCB. PMID- 18384630 TI - Renal cell carcinoma: what does the future hold? PMID- 18384631 TI - Accuracy of life tables in predicting overall survival in patients after radical prostatectomy. AB - OBJECTIVE: To test the accuracy of life tables (LT), the standard tool for predicting life-expectancy (LE), but the accuracy of which is unknown in patients with prostate cancer, where the 10-year LE is a widely accepted threshold for the delivery of definitive therapy. PATIENTS AND METHODS: We tested the accuracy of predictions of LE from LT in 9678 men treated with radical prostatectomy (RP) for prostate cancer. The predictions of LE from LT at 10 years after RP were compared to Kaplan Meier-derived 10-year survival values. Moreover, the accuracy of LT predictions was quantified in a Cox-regression using Harrell's concordance index. To control for the effect of prostate cancer mortality, analyses were repeated in a subset of 5955 patients with no evidence of disease recurrence. Additional stratification schemes were applied to control for age and comorbidity. RESULTS: At RP, the median age was 64 years, the median Charlson Comorbidity Index (CCI) was 1 and the median LT-derived LE was 16 years. The median actuarial survival was not reached (mean 12.4 years). In the whole group the LT-predicted 10-year survival was 96.8%, vs an observed of 75.3%. In men with no disease recurrence the LT-predicted survival was 97.3%, vs 81.1% observed. After age and CCI stratification, LT overestimated the 10-year survival the most in those aged 65 69 years and in patients with CCI scores of >2. CONCLUSION: The overestimation of LE can lead to overtreatment of prostate cancer, especially in those men who die early from other causes. PMID- 18384632 TI - Is there an alternative to pad tests? Correlation of subjective variables of severity of urinary loss to the 1-h pad test in women with stress urinary incontinence. AB - OBJECTIVE: To compare the 1-h pad test in women who have urodynamically confirmed stress incontinence (USI) with a patient-based 3-point symptom severity scale and validated quality of life (QoL) questionnaires. PATIENTS AND METHODS: In all, 98 women with USI were prospectively recruited; all had a 1-h pad test and completed the validated disease-specific QoL questionnaires, including short forms of the International Consultation on Incontinence Questionnaire (ICIQ-SF), Urogenital Distress Inventory (UDI-6), and Incontinence Impact Questionnaire (IIQ-7). In addition, the severity of incontinence was determined using the Stamey grading scale, a visual analogue scale (VAS) score, and a patient-based 3-point symptom severity scale. RESULTS: The VAS, symptom severity scale, Stamey grade, UDI-6, and IIQ-7 failed to correlate significantly with the 1-h pad test. Only the ICIQ SF correlated significantly with this test. CONCLUSION: The ICIQ-SF is easy to administer and, in this study, correlated best with the 1-h pad test in women with pure primary or secondary USI. It incorporates both symptom severity and QoL variables. We recommend its routine use in clinical practice. PMID- 18384633 TI - Disabling complications with slings for managing female stress urinary incontinence. AB - OBJECTIVE: To report an increase in the referral of patients with disabling complications after the failure of conservative therapy, their presentation, final surgical management and clinical outcome, following the use of non autologous slings (NAS), currently the primary surgical procedure for managing stress urinary incontinence (SUI) in women. PATIENT AND METHODS: Thirty-eight patients (mean age 64 years) required surgical management for disabling complications after placing a NAS for SUI. Sling types were synthetic (25), xenografts (six) and allografts (four). Twenty (53%) patients presented with bladder outlet obstruction, 13 (34%) with sling erosion, three (8%) with worsened SUI, and two (5%) with unobstructive severe urgency and frequency. RESULTS: The sling was dissected and incised with no complication in 19 of 20 patients. One had a posterior urethral defect during sling dissection. Twelve patients (60%) acquired normal voiding and were continent. Among the 13 patients who had the sling dismantled and urethrolysis, two had recurrent or persistent SUI, two de novo urgency/frequency and one developed osteitis pubis. Three patients with disabling SUI received a pubovaginal sling placed proximal to the bladder neck, and had an overall improvement in their urinary control with no retention. Two unobstructed patients with urgency and frequency did not improve with anticholinergic medication and pelvic floor therapy, and are now candidates for botulinum toxin injection or neurostimulation. CONCLUSIONS: The complication rate with periurethral NAS for managing SUI in females is substantial. Patients with refractory urgency/frequency after the sling need a complete evaluation with cystoscopy and video-urodynamics. Obstruction and erosion are the commonest problems and require surgical correction. PMID- 18384634 TI - Comparison of prostate-specific antigen recurrence-free survival in a contemporary cohort of patients undergoing either radical retropubic or robot assisted laparoscopic radical prostatectomy. AB - OBJECTIVES: To compare the prostate-specific antigen (PSA) recurrence (PSAR) rates in patients undergoing robot-assisted laparoscopic radical prostatectomy (RALP) or radical retropubic prostatectomy (RRP). PATIENTS AND METHODS: Data from 797 consecutive patients who had RALP or RRP between August 2003 and January 2007 were retrieved from our database. Age, race, body mass index, PSA level, estimated blood loss (EBL), clinical and pathological stage, biopsy and pathological Gleason score, lymph node involvement, positive surgical margin (PSM) status, and prostate weight were compared between the groups. Multivariate analysis (logistic and Cox regression) was used to adjust for differences in clinical and pathological features when comparing the risk for PSM and PSAR. RESULTS: In all, 362 men had RALP and 435 had RRP; the mean follow-up was 1.09 and 1.37 years, respectively. RALP patients had a significantly lower clinical stage, Gleason score and EBL (P < 0.001). There was no significant difference in PSM between RALP and RRP in univariate (P = 0.701) and multivariate analyses (P = 0.095). The risk of PSAR for patients undergoing RALP or RRP was not significantly different after adjusting for clinical (hazard ratio 0.82, 95% confidence interval 0.48-1.38; P = 0.448) and pathological differences (0.94, 0.55-1.61; P = 0.824). CONCLUSIONS: Patients undergoing RALP had a lower EBL and lower-risk disease. After adjusting for differences in clinical and pathological features, there was no significant difference in early PSAR between patients undergoing RALP or RRP. PMID- 18384635 TI - Immunohistochemical estimation of hypoxia in human obstructed bladder and correlation with clinical variables. AB - OBJECTIVE: To investigate the tissue distribution of ischaemia in human detrusor in patients with bladder outlet obstruction (BOO) and to correlate the results with clinical variables, as clinical BOO is a common problem in ageing men and ischaemia might be important in detrusor dysfunction. PATIENTS AND METHODS: From September 2004 to October 2006, 70 patients were recruited, comprising 60 scheduled for surgery to treat benign prostatic hyperplasia (the study group) and 10 as controls. Detrusor tissue was retrieved and stained for hypoxia-inducible factor (HIF)-1alpha, a cellular marker of hypoxia. RESULTS: The mean (sd) total number of cells immunoreactive to HIF-1alpha in the study group was 93.3 (48.09), and in the specimens from the control group only few rare cells showed weak immunoreactivity to HIF-1alpha (0-2). Positive cells were in different proportions between muscle bundles and submucosa, expressed mainly in stromal cells. The urothelium and detrusor muscle showed no immunoreactivity to HIF 1alpha. There was strong immunoreactivity in patients with prolonged BOO (<10 years), declining thereafter, and in those patients with urinary retention. CONCLUSIONS: The urothelium and detrusor seem to be more resistant to hypoxic stress, while stromal cells perceive low oxygen tension. The bladder response to chronic hypoxia through HIF-1alpha expression is limited in time and might depend on the functional status of the detrusor. PMID- 18384636 TI - Quantifying mast cells in bladder pain syndrome by immunohistochemical analysis. AB - OBJECTIVES: To evaluate a simple method for counting mast cells, thought to have a role in the pathophysiology of bladder pain syndrome (BPS, formerly interstitial cystitis, a syndrome of pelvic pain perceived to be related to the urinary bladder and accompanied by other urinary symptoms, e.g. frequency and nocturia), as >28 mast cells/mm(2) is defined as mastocytosis and correlated with clinical outcome. PATIENTS AND METHODS: The current enzymatic staining method (naphtolesterase) on 10 microm sections for quantifying mast cells is complicated. In the present study, 61 patients had detrusor biopsies taken between 2002 and 2005; the patients were given a clinical score, and sections of the biopsy stained with (i) naphtolesterase on 10 microm sections, staining every third section, or (ii) immunohistochemically with antitryptase on both 10 microm and 3 microm sections, with two and six unstained sections between, respectively. Mast cells were counted according to a well-defined procedure. RESULTS: The old and the new methods, on 10 and 3 microm sections, showed a good correlation between mast cell counts. When using tryptase staining and 3 microm sections, the mast cell number correlated well with the clinical score (Spearman's rho 0.576; 95% confidence interval 0.155-0.820) and 27 mast cells/mm(2) was the threshold suggesting mastocytosis. CONCLUSIONS: We recommend taking biopsies from the detrusor of patients with suspected BPS and examining them with tryptase-stained 3 microm thick sections, with every seventh section used for quantification; 27 mast cells/mm(2) is considered indicative of mastocytosis. PMID- 18384637 TI - Paclitaxel and cisplatin as intravesical agents against non-muscle-invasive bladder cancer. AB - OBJECTIVES: To investigate the effects of cisplatin and paclitaxel against human bladder cancer cells in vitro, and to obtain both pharmacokinetic and pharmacodynamic data after intravesical administration in mice. MATERIALS AND METHODS: Six bladder cancer cell lines (J82, KU7, RT4, SW780, T24, UMUC3) were treated with various combined doses of both drugs and cell proliferation was evaluated 3 days later. In vivo, solutions of cisplatin and micellar paclitaxel were instilled transurethrally in female mice and pharmacokinetic data were acquired using high-performance liquid chromatography-mass spectrometry and atomic absorption methods. To obtain efficacy data, mice with orthotopic KU7-luc tumours were administered cisplatin and/or micellar paclitaxel intravesically, and the tumour burden quantified using bioluminescence imaging. RESULTS: In vitro, both cisplatin and paclitaxel potently decreased the proliferation of all cell lines tested, and in combination had an additive but not a synergistic effect. After intravesical instillation, mouse serum concentrations of cisplatin and paclitaxel were in the low microgram/millilitre range and bladder tissue concentrations achieved were 82 and 241 microg/g, respectively. Similar drug levels were reached using combined therapy. In vivo, all chemotherapeutic agents significantly inhibited bladder tumour growth, with the best results for combined therapy and micellar paclitaxel alone. However, there was toxicity in the combined treatment arm. CONCLUSIONS: Both cisplatin and paclitaxel were absorbed at effective amounts into bladder tissues. As intravesical agents, paclitaxel had slightly stronger anticancer potency than cisplatin. Due to increased adverse events, caution should be exercised when combining both cisplatin and paclitaxel intravesically. PMID- 18384638 TI - Robotically assisted radical cystectomy. PMID- 18384639 TI - Twenty years of experience with Krzeski's cystovaginoplasty for vaginal agenesis in Mayer-Rokitansky-Kuster-Hauser syndrome: anatomical, histological, cytological and functional results. AB - OBJECTIVE: To evaluate the long-term anatomical results using the original method of vaginal reconstruction with a pedicled bladder flap (Krzeski's cystovaginoplasty, CVP) in women with Mayer-Rokitansky-Kuster-Hauser syndrome (MRKHS) and the evaluation of radiological, histological, cytohormonal and functional results of CVP. PATIENTS AND METHODS: Between 1981 and 2000, 38 patients (mean age 22.5 years, range 18-40) with MRKHS underwent CVP. A physical examination was used to evaluate the anatomical results, 27 patients had vaginography, and biopsies of the neovaginal wall and cytohormonal smears were taken in two. Functional sexual and urinary results, and opinions on CVP, were evaluated by an inventory mailed to 37 patients in 2000. RESULTS: The anatomical result was good in 37 patients during a mean (range) follow-up of 9 (0.25-19) years; the result was good in 30 patients, but seven developed vaginal stenosis that was successfully repaired. There were two cases of post-coital vesicovaginal fistula (VVF) at 18 months after CVP. Vaginal biopsies showed epithelialization of the posterior vaginal wall and gradual metaplastic changes from urothelium to stratified nonsquamous epithelium. Cytological smears showed a normal biphasic pattern and neovaginal susceptibility to hormonal milieu. In all, 27 patients (73%) responded to the questionnaire. All had sexual partners and started sexual intercourse at a mean of 14 months after CVP; 89% experience orgasms and in 48% the vagina was the source; 40% sometimes used lubricants and seven (26%) used vaginal dilators. Lower urinary tract symptoms (LUTS) after CVP were reported by 19 (66%) of the women and in six the LUTS were persistent. One patient was not satisfied with the functional and anatomical result of CVP, 89% declared that it improved sexual life, 93% would undergo CVP again and in 92% the quality of their sexual life was improved. All patients, when asked, stated that they would recommend CVP to another patient with MRKHS. CONCLUSIONS: Vaginal reconstruction by CVP is characterized by good anatomical and functional results, sustained by long-term observation. The resultant epithelium is very similar to that of the native vagina in histology and function. All vaginas are functional and the level of patient satisfaction was high. The level of complications was acceptable, but in some patients LUTS can persist. PMID- 18384640 TI - Human immunodeficiency virus-associated prostate cancer: clinicopathological findings and outcome in a multi-institutional study. AB - OBJECTIVE: To characterize the clinicopathological findings and the outcome of human immunodeficiency virus (HIV)-infected patients diagnosed and treated for prostate carcinoma, as HIV-positive men being treated with highly active antiretroviral therapy (HAART) are living longer and thus are more likely to develop cancers such as prostate cancer. PATIENTS AND METHODS: We performed a retrospective, multi-institutional study involving HIV-positive men with concomitant prostate carcinoma. We collected data regarding patient demographics (age, race), HIV status (CD4(+) cell count, HIV viral load, HAART), PSA level (at cancer diagnosis), symptoms and signs, radiological findings, pathology (Gleason score, stage), cancer treatment (type, side-effects), and outcome (response, survival). Accrued data was analysed using descriptive statistics. RESULTS: We identified 17 patients (mean age 59 years) with HIV-associated prostate adenocarcinoma. The mean CD4(+) count was 336 cells/mm(3) and the mean HIV viral load was 17 319 copies/mL. In all, 14 (82%) of these men were receiving HAART. Most patients were diagnosed with carcinoma after an abnormal screening PSA level. The mean PSA level was 30 ng/mL. Only six (35%) men had an abnormal prostate on examination. The mean Gleason score was 6.8, and in most cases, cancer was confined to the prostate gland. Most patients were amenable to curative treatment with hormonal therapy, radiation, and/or prostatectomy. There were no serious treatment related side-effects. One patient remained untreated. All treated patients had a complete response (undetectable PSA level). Most patients were long-term survivors. Documented death in five cases was unrelated to prostate cancer. CONCLUSION: The management of HIV-positive men with prostate carcinoma in the HAART era is becoming increasingly important. Our data shows that in men receiving HAART, their age, PSA levels, clinical presentation, management, and outcome from treated prostate carcinoma does not appear to be significantly altered by HIV status. Therefore, we recommend that patients with prostate cancer and well-controlled HIV viraemia be managed similarly to their HIV-negative counterparts. PMID- 18384641 TI - Toxicity and endocytosis of spinocerebellar ataxia type 6 polyglutamine domains: role of myosin IIb. AB - Spinocerebellar ataxia type 6 (SCA6) is a dominantly inherited neurodegenerative disease caused by a small expansion of CAG repeats in the sequence coding for the cytoplasmic C-terminal region of the Ca(v)2.1 subunit of P/Q-type calcium channels. We have tested the toxicity of mutated Ca(v)2.1 C-terminal domains expressed in the plasma membrane. In COS-7 cells, CD4-green fluorescent protein fused to Ca(v)2.1 C-terminal domains containing expanded 24 polyglutamine (Q) tracts displayed increased toxicity and stronger expression at the cell surface relative to 'normal' 12 Q tracts, partially because of reduced endocytosis. Glutathione S-transferase pull-down and proteomic analysis indicated that Ca(v)2.1 C-termini interact with the heavy and light chains of cerebellar myosin IIB, a molecular motor protein. This interaction was confirmed by coimmunoprecipitation from rat cerebellum and COS-7 cells and shown to be direct by binding of in vitro-translated (35)S-myosin IIB heavy chain. In COS-7 cells, incremented polyglutamine tract length increased the interaction with myosin IIB. Furthermore, the myosin II inhibitor blebbistatin reversed the effects of polyglutamine expansion on plasma membrane expression. Our findings suggest a key role of myosin IIB in promoting accumulation of mutant Ca(v)2.1Ct at the plasma membrane and suggest that this gain of function might contribute to the pathogenesis of SCA6. PMID- 18384642 TI - Neuronal pigmented autophagic vacuoles: lipofuscin, neuromelanin, and ceroid as macroautophagic responses during aging and disease. AB - The most striking morphologic change in neurons during normal aging is the accumulation of autophagic vacuoles filled with lipofuscin or neuromelanin pigments. These organelles are similar to those containing the ceroid pigments associated with neurologic disorders, particularly in diseases caused by lysosomal dysfunction. The pigments arise from incompletely degraded proteins and lipids principally derived from the breakdown of mitochondria or products of oxidized catecholamines. Pigmented autophagic vacuoles may eventually occupy a major portion of the neuronal cell body volume because of resistance of the pigments to lysosomal degradation and/or inadequate fusion of the vacuoles with lysosomes. Although the formation of autophagic vacuoles via macroautophagy protects the neuron from cellular stress, accumulation of pigmented autophagic vacuoles may eventually interfere with normal degradative pathways and endocytic/secretory tasks such as appropriate response to growth factors. PMID- 18384643 TI - Steroid-induced sexual differentiation of the developing brain: multiple pathways, one goal. AB - Hormone exposure, including testosterone and its metabolite estradiol, induces a myriad of effects during a critical period of brain development that are necessary for brain sexual differentiation. Nuclear volume, neuronal morphology, and astrocyte complexity are examples of the wide range of effects by which testosterone and estradiol can induce permanent changes in the function of neurons for the purpose of reproduction in adulthood. This review will examine the multitude of mechanisms by which steroid hormones induce these permanent changes in brain structure and function. Elucidating how steroids alter brain development sheds light on how individual variation in neuronal phenotype is established during a critical period. PMID- 18384644 TI - Retrograde axonal transport and motor neuron disease. AB - Transport of material between extensive neuronal processes and the cell body is crucial for neuronal function and survival. Growing evidence shows that deficits in axonal transport contribute to the pathogenesis of multiple neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). Here we review recent data indicating that defects in dynein-mediated retrograde axonal transport are involved in ALS etiology. We discuss how mutant copper-zinc superoxide dismutase (SOD1) and an aberrant interaction between mutant SOD1 and dynein could perturb retrograde transport of neurotrophic factors and mitochondria. A possible contribution of axonal transport to the aggregation and degradation processes of mutant SOD1 is also reviewed. We further consider how the interference with axonal transport and protein turnover by mutant SOD1 could influence the function and viability of motor neurons in ALS. PMID- 18384646 TI - P2Y receptor mediated inhibitory modulation of noradrenaline release in response to electrical field stimulation and ischemic conditions in superfused rat hippocampus slices. AB - In this study, the inhibitory regulation of the release of noradrenaline (NA) by P2 receptors was investigated in hippocampus slices pre-incubated with [(3)H]NA. Electrical field stimulation (EFS; 2 Hz, 240 shocks, and 1 ms) released NA in an outside [Ca(2+)]-dependent manner, and agonists of P2Y receptors inhibited the EFS-evoked [(3)H]NA release with pharmacological profile similar to that of the P2Y(1) and P2Y(13) receptor subtypes. This inhibitory modulation was counteracted by bicuculline and 6-cyano-2,3-dihydroxy-7-nitro-quinoxaline + 2-amino-5 phosphonovalerate and 2-amino-4-phosphonobutyrate. In contrast, the excess release in response to 30 min combined oxygen and glucose deprivation was outside [Ca(2+)] independent, but still sensitive to the inhibition of both facilitatory P2X(1) and inhibitory P2Y(1) receptors. Whereas mRNA encoding P2Y(12) and P2Y(13) receptor subunits were expressed in the brainstem, P2Y(1) receptor immunoreactivity was localized to neuronal somata and dendrites innervated by the mossy fiber terminals in the CA3 region of the hippocampus, as well as somata of granule cells and interneurons in the dentate gyrus. In summary, in addition to the known facilitatory modulation via P2X receptors, EFS-evoked [(3)H]NA outflow in the hippocampus is subject to inhibitory modulation by P2Y(1)/P2Y(13) receptors. Furthermore, endogenous activation of both facilitatory and inhibitory P2 receptors may participate in the modulation of pathological NA release under ischemic-like conditions. PMID- 18384645 TI - Changes in endoplasmic reticulum stress proteins and aldolase A in cells exposed to dopamine. AB - In Parkinson's disease, oxidative stress is implicated in protein misfolding and aggregation, which may activate the unfolded protein response by the endoplasmic reticulum (ER). Dopamine (DA) can initiate oxidative stress via H(2)O(2) formation by DA metabolism and by oxidation into DA quinone. We have previously shown that DA quinone induces oxidative protein modification, mitochondrial dysfunction in vitro, and dopaminergic cell toxicity in vivo and in vitro. In this study, we used cysteine- and lysine-reactive fluorescent dyes with 2D difference in-gel electrophoresis, mass spectrometry, and peptide mass fingerprint analysis to identify proteins in PC12 cell mitochondrial-enriched fractions that were altered in abundance following DA exposure (150 muM, 16 h). Quantitative changes in proteins labeled with fluorescent dyes indicated increases in a subset of proteins after DA exposure: calreticulin, ERp29, ERp99, Grp58, Grp78, Grp94 and Orp150 (149-260%), and decreased levels of aldolase A (39 42%). Changes in levels of several proteins detected by 2D difference in-gel electrophoresis were confirmed by western blot. Using this unbiased proteomics approach, our findings demonstrated that in PC12 cells, DA exposure leads to a cellular response indicative of ER stress prior to the onset of cell death, providing a potential link between DA and the unfolded protein response in the pathogenesis of Parkinson's disease. PMID- 18384648 TI - Alpha2 adrenoreceptor agonist regulates protein kinase C-induced heat shock protein 27 phosphorylation in C6 glioma cells. AB - Dexmedetomidine (Dexmd), a potent and highly specific alpha(2) adrenoreceptor agonist, is an efficient therapeutic agent for sedation. Dexmd has been recently reported to have a neuroprotective effect. Heat shock protein (HSP) 27, a low molecular weight HSP has been shown to be expressed following cerebral ischemia in astrocytes but not in neurons. HSP27 expression is involved in ischemic tolerance of the brain. This study investigated the effect of Dexmd on HSP27 in rat C6 glioma cells. 12-O-tetradecanoylphorbol-13-actate (TPA), a direct activator of protein kinase C (PKC), stimulated the phosphorylation of HSP27 at Ser82, but not Ser15 in a time-dependent manner. Prostaglandin (PG) E(1) or PGE(2) which activates the adenylyl cyclase-cAMP system as well as forskolin and dibutyryl-cAMP, suppressed the TPA-induced phosphorylation of HSP27. Dexmd reversed the suppression of HSP27 phosphorylation by the adenylyl cyclase-cAMP system. Therefore, these results strongly suggest that Dexmd reverses the suppression of HSP27 phosphorylation by the adenylyl cyclase-cAMP system activation through the inhibition of its system in C6 cells. alpha(2) Adrenoreceptor agonists may therefore show a neuroprotective effect through the modification of HSP27 phosphorylation induced by PKC activation. PMID- 18384647 TI - Permeation of blood-borne IL15 across the blood-brain barrier and the effect of LPS. AB - Interleukin15 (IL 15) is a proinflammatory cytokine with elevated concentrations in autoimmune diseases involving the periphery (e.g. rheumatoid arthritis) and CNS (e.g. multiple sclerosis). Its interactions with the blood-brain barrier (BBB) were studied in normal and lipopolysaccharide (LPS)-treated mice. (125)I IL15 remained intact for at least 10 min after i.v. injection and reached CNS parenchyma with regional differences between brain and spinal cord. Both in vivo and in situ brain perfusion of (125)I-IL15 showed that its permeation of the BBB was non-saturable. LPS induced a significant increase of IL15 uptake by the brain and spinal cord, partly related to a higher general permeability of the BBB. The results suggest that the BBB is an interface for blood-borne IL15 to interact with the CNS in the basal state and during inflammation. PMID- 18384649 TI - PPAR: a therapeutic target in Parkinson's disease. AB - Parkinson's disease (PD) is a progressive and chronic neurodegenerative disorder, characterized by progressive loss of dopaminergic neurons in substantia nigra. The etiology and pathogenesis of PD is still elusive, however, a large body of evidence suggests a prominent role of oxidative stress, inflammation, apoptosis, mitochondrial dysfunction and proteosomal dysfunction in the pathogenesis of PD. Due to multifactorial nature of the disease, currently available drug therapy cannot halt / slow down the disease progression, and only provides symptomatic relief. Peroxisome proliferator-activated receptor (PPAR), a member of nuclear receptor superfamily, regulates development, tissue differentiation, inflammation, mitochondrial function, wound healing, lipid metabolism and glucose metabolism. Recently, several PPAR agonists were shown to exert neuroprotective activity against oxidative damage, inflammation and apoptosis in several neurodegenerative disorders including Alzheimer's, Parkinson's, Huntington's, amyotrophic lateral sclerosis and multiple sclerosis. Similarly, regular intake of PPAR activating non-steroidal anti-inflammatory drugs such as indomethacin and ibuprofen was associated with reduced incidence and progression of neurodegenerative disorders in several epidemiological studies. In this article, we review studies relating to the neuroprotective effect of PPAR agonists in in vitro and in vivo models of PD. Similarly, the pharmacological mechanism in neuroprotective actions of PPAR agonists is also reviewed. In conclusion, PPAR agonists exert neuroprotective actions by regulating the expression of a set of genes involved in cell survival processes, and could be a therapeutic target in debilitating neurodegenerative illnesses such as PD. PMID- 18384650 TI - Inflammatory events in hippocampal slice cultures prime neuronal susceptibility to excitotoxic injury: a crucial role of P2X7 receptor-mediated IL-1beta release. AB - We investigated the consequences of transient application of specific stimuli mimicking inflammation to hippocampal tissue on microglia activation and neuronal cell vulnerability to a subsequent excitotoxic insult. Two-week-old organotypic hippocampal slice cultures, from 7-day-old C57BL/6 donor mice, were exposed for 3 h to lipopolysaccharide (LPS; 10 ng/mL) followed by 3 h co-incubation with 1 mM ATP, or 100 microM 2'3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate triethylammonium, a selective P2X(7) receptor agonist. These treatments in combination, but not individually, induced a pronounced activation and apoptotic like death of macrophage antigen-1 (MAC-1)-positive microglia associated with a massive release of interleukin (IL)-1beta exceeding that induced by LPS alone. Antagonists of P2X(7) receptors prevented these effects. Transient pre-exposure of slice cultures to a combination of LPS and P2X(7) receptor agonists, but not either one or the other alone, significantly exacerbated CA3 pyramidal cell loss induced by subsequent 12 h exposure to 8 microM alpha-amino-3-hydroxy-5-methyl-4 isoxazole propinate (AMPA). Potentiation of AMPA toxicity was prevented when IL 1beta production or its receptor signaling were blocked by an inhibitor of interleukin-converting-enzyme or IL-1 receptor antagonist during application of LPS + ATP. The same treatments did not prevent microglia apoptosis-like death. These findings show that transient exposure to specific pro-inflammatory stimuli in brain tissue can prime neuronal susceptibility to a subsequent excitotoxic insult. P2X(7) receptor stimulation, and the consequent IL-1beta release, is mandatory for exacerbation of neuronal loss. These mechanisms may contribute to determine cell death/survival in acute and chronic neurodegenerative conditions associated with inflammatory events. PMID- 18384652 TI - Acquired von Willebrand disease: potential contribution of the von Willebrand factor collagen-binding to the identification of functionally inhibiting auto antibodies to von Willebrand factor: a rebuttal. PMID- 18384651 TI - A first-in-human phase I trial of locally delivered human plasmin for hemodialysis graft occlusion. AB - BACKGROUND: Hemodialysis (HD) grafts often fail because of stenosis at the venous anastomosis and thrombotic occlusion. Percutaneous management relies on thrombolysis with plasminogen activators, mechanical removal of thrombus, and angioplasty of the stenotic lesion. OBJECTIVES: This report describes a phase I trial using Plasmin (Human) TAL 05-00018, a direct-acting fibrinolytic agent, to evaluate safety and, secondarily, to establish effective thrombolytic dosing. PATIENTS/METHODS: Six cohorts of five patients with acute HD graft occlusion documented by angiography were treated with escalating dosages of plasmin (1, 2, 4, 8, 12, and 24 mg) infused over 30 min via criss-crossed pulse-spray catheters within the graft. The primary efficacy endpoint was > or =50% thrombolysis, as determined by comparison of pre-plasmin and 30-min post-plasmin fistulograms. RESULTS: Of 31 subjects who received study drug (safety population), one withdrew and 30 completed the trial (evaluable for efficacy). There was no significant change in plasma alpha-2 antiplasmin or fibrinogen concentration, major bleeding did not occur, and there were no deaths. Serious adverse events in four patients were not related to the study drug. There was a dose-response relationship for the primary efficacy endpoint, all five subjects receiving 24 mg achieving >75% lysis. CONCLUSIONS: This first phase I study of Plasmin (Human) TAL 05-00018, infused into thrombosed HD grafts, documents safety at dosages of 1-24 mg and an effective thrombolytic dosage of 24 mg. The results establish a foundation for further clinical study of catheter-based plasmin administration in thrombotic disorders. PMID- 18384654 TI - Evolutionary dynamics of a sexual ornament in the house sparrow (Passer domesticus): the role of indirect selection within and between sexes. AB - The relative contribution of sexual and natural selection to evolution of sexual ornaments has rarely been quantified under natural conditions. In this study we used a long-term dataset of house sparrows in which parents and offspring were matched genetically to estimate the within- and across-sex genetic basis for variation and covariation among morphological traits. By applying two-sex multivariate "animal models" to estimate genetic parameters, we estimated evolutionary changes in a male sexual ornament, badge size, from the contribution of direct and indirect selection on correlated traits within males and females, after accounting for overlapping generations and age-structure. Indirect natural selection on genetically correlated traits in males and females was the major force causing evolutionary change in the male ornament. Thus, natural selection on female morphology may cause indirect evolutionary changes in male ornaments. We observed however no directional phenotypic change in the ornament size of one year-old males during the study period. On the other hand, changes were recorded in other morphological characters of both sexes. Our analyses of evolutionary dynamics in sexual characters require application of appropriate two-sex models to account for how selection on correlated traits in both sexes affects the evolutionary outcome of sexual selection. PMID- 18384655 TI - Disassortative mating, sexual specialization, and the evolution of gender dimorphism in heterodichogamous Acer opalus. AB - In sexually polymorphic species, the morphs are maintained by frequency-dependent selection through disassortative mating. In heterodichogamous populations in which disassortative mating occurs between the protandrous and protogynous morphs, a decrease in female fitness in one morph is hypothesized to drive sexual specialization in the other morph, resulting in dimorphic populations. We test these ideas in a population of the heterodichogamous species, Acer opalus. We assessed both prospective gender of individuals in terms of their allocations and actual parentage using microsatellites; we found that most matings in A. opalus occur disassortatively. We demonstrate that the protogynous morph is maintained by frequency-dependent selection, but that maintenance of males versus protandrous individuals depends on their relative siring success, which changes yearly. Seeds produced later in the reproductive season were smaller than those produced earlier; this should compromise reproduction through ovules in protandrous individuals, rendering them male biased in gender. Time-dependent gender and paternity analyses indicate that the sexual morphs are specialized in their earlier sexual functions, mediated by the seasonal decrease in seed size. Our results confirm that mating patterns are context-dependent and change seasonally, suggesting that sexual specialization can be driven by seasonal effects on fitness gained through one of the two sexual functions. PMID- 18384656 TI - Increased postcopulatory sexual selection reduces the intramale variation in sperm design. AB - Sperm competition is an important force driving the evolution of sperm design and function. Inter- and intraspecific variation in sperm design are strongly influenced by the risk of sperm competition in many taxa. In contrast, the variation among sperm of one male (intramale variation) is less well understood. We investigated intramale variation in sperm design in passerine birds and found that risk of sperm competition is negatively associated with intramale variation. This result is the first clear evidence that variation among sperm within an individual male is influenced by postcopulatory sexual selection. Our finding has important implications for male traits under pre- and postcopulatory sexual selection. PMID- 18384657 TI - Phylogenetic analysis of the ecology and evolution of mammalian sleep. AB - The amount of time asleep varies greatly in mammals, from 3 h in the donkey to 20 h in the armadillo. Previous comparative studies have suggested several functional explanations for interspecific variation in both the total time spent asleep and in rapid-eye movement (REM) or "quiet" (non-REM) sleep. In support of specific functional benefits of sleep, these studies reported correlations between time in specific sleep states (NREM or REM) and brain size, metabolic rate, and developmental variables. Here we show that estimates of sleep duration are significantly influenced by the laboratory conditions under which data are collected and that, when analyses are limited to data collected under more standardized procedures, traditional functional explanations for interspecific variation in sleep durations are no longer supported. Specifically, we find that basal metabolic rate correlates negatively rather than positively with sleep quotas, and that neither adult nor neonatal brain mass correlates positively with REM or NREM sleep times. These results contradict hypotheses that invoke energy conservation, cognition, and development as drivers of sleep variation. Instead, the negative correlations of both sleep states with basal metabolic rate and diet are consistent with trade-offs between sleep and foraging time. In terms of predation risk, both REM and NREM sleep quotas are reduced when animals sleep in more exposed sites, whereas species that sleep socially sleep less. Together with the fact that REM and NREM sleep quotas correlate strongly with each other, these results suggest that variation in sleep primarily reflects ecological constraints acting on total sleep time, rather than the independent responses of each sleep state to specific selection pressures. We propose that, within this ecological framework, interspecific variation in sleep duration might be compensated by variation in the physiological intensity of sleep. PMID- 18384658 TI - The effects of multiple infections on the expression and evolution of virulence in a Daphnia-endoparasite system. AB - Multiple infections of a host by different strains of the same microparasite are common in nature. Although numerous models have been developed in an attempt to predict the evolutionary effects of intrahost competition, tests of the assumptions of these models are rare and the outcome is diverse. In the present study we examined the outcome of mixed-isolate infections in individual hosts, using a single clone of the waterflea Daphnia magna and three isolates of its semelparous endoparasite Pasteuria ramosa. We exposed individual Daphnia to single- and mixed-isolate infection treatments, both simultaneously and sequentially. Virulence was assessed by monitoring host mortality and fecundity, and parasite spore production was used as a measure of parasite fitness. Consistent with most assumptions, in multiply infected hosts we found that the virulence of mixed infections resembled that of the more virulent competitor, both in simultaneous multiple infections and in sequential multiple infections in which the virulent isolate was first to infect. The more virulent competitor also produced the vast majority of transmission stages. Only when the less virulent isolate was first to infect, the intrahost contest resembled scramble competition, whereby both isolates suffered by producing fewer transmission stages. Surprisingly, mixed-isolate infections resulted in lower fecundity-costs for the hosts, suggesting that parasite competition comes with an advantage for the host relative to single infections. Finally, spore production correlated positively with time-to-host-death. Thus, early-killing of more competitive isolates produces less transmission stages than less virulent, inferior isolates. Our results are consistent with the idea that less virulent parasite lines may be replaced by more virulent strains under conditions with high rates of multiple infections. PMID- 18384659 TI - Rapid speciation following recent host shifts in the plant pathogenic fungus Rhynchosporium. AB - Agriculture played a significant role in increasing the number of pathogen species and in expanding their geographic range during the last 10,000 years. We tested the hypothesis that a fungal pathogen of cereals and grasses emerged at the time of domestication of cereals in the Fertile Crescent and subsequently speciated after adaptation to its hosts. Rhynchosporium secalis, originally described from rye, causes an important disease on barley called scald, although it also infects other species of Hordeum and Agropyron. Phylogenetic analyses based on four DNA sequence loci identified three host-associated lineages that were confirmed by cross-pathogenicity tests. Bayesian analyses of divergence time suggested that the three lineages emerged between approximately 1200 to 3600 years before present (B.P.) with a 95% highest posterior density ranging from 100 to 12,000 years B.P. depending on the implemented clock models. The coalescent inference of demographic history revealed a very recent population expansion for all three pathogens. We propose that Rhynchosporium on barley, rye, and Agropyron host species represent three cryptic pathogen species that underwent independent evolution and ecological divergence by host-specialization. We postulate that the recent emergence of these pathogens followed host shifts. The subsequent population expansions followed the expansion of the cultivated host populations and accompanying expansion of the weedy Agropyron spp. found in fields of cultivated cereals. Hence, agriculture played a major role in the emergence of the scald diseases, the adaptation of the pathogens to new hosts and their worldwide dissemination. PMID- 18384660 TI - Host-specific toxins: effectors of necrotrophic pathogenicity. AB - Host-specific toxins (HSTs) are defined as pathogen effectors that induce toxicity and promote disease only in the host species and only in genotypes of that host expressing a specific and often dominant susceptibility gene. They are a feature of a small but well-studied group of fungal plant pathogens. Classical HST pathogens include species of Cochliobolus, Alternaria and Pyrenophora. Recent studies have shown that Stagonospora nodorum produces at least four separate HSTs that interact with four of the many quantitative resistance loci found in the host, wheat. Rationalization of fungal phylogenetics has placed these pathogens in the Pleosporales order of the class Dothideomycetes. It is possible that all HST pathogens lie in this order. Strong evidence of the recent lateral gene transfer of the ToxA gene from S. nodorum to Pyrenophora tritici-repentis has been obtained. Hallmarks of lateral gene transfer are present for all the studied HST genes although definitive proof is lacking. We therefore suggest that the Pleosporales pathogens may have a conserved propensity to acquire HST genes by lateral transfer. PMID- 18384661 TI - Antibody action after phagocytosis promotes Cryptococcus neoformans and Cryptococcus gattii macrophage exocytosis with biofilm-like microcolony formation. AB - Antibody-mediated phagocytosis was discovered over a century ago but little is known about antibody effects in phagolysosomes. We explored the consequences of antibody-mediated phagocytosis for two closely related human pathogenic fungal species, Cryptococcus neoformans and Cryptococcus gattii, of which C. neoformans encompasses two varieties: neoformans and grubii. The interaction between C. neoformans varieties grubii and neoformans and host cells has been extensively studied, but that of C. gattii and macrophages remains largely unexplored. Like C. neoformans, antibody-mediated phagocytosis of C. gattii cells was followed by intracellular replication, host cell cytoplasmic polysaccharide accumulation and phagosomal extrusion. Both C. gattii and C. neoformans cells exited macrophages in biofilm-like microcolonies where the yeast cells were aggregated in a polysaccharide matrix that contained bound antibody. In contrast, complement opsonized C. neoformans variety grubii cells were released from macrophages dispersed as individual cells. Hence, both antibody- and complement-mediated phagocytosis resulted in intracellular replication but the mode of opsonization affected the outcome of exocytosis. The biofilm-like microcolony exit strategy of C. neoformans and C. gattii following antibody opsonization reduced fungal cell dispersion. This finding suggests that antibody agglutination effects persist in the phagosome to entangle nascent daughter cells and this phenomenon may contribute to antibody-mediated protection. PMID- 18384662 TI - Clinical trial design in adult reflux disease: a methodological workshop. AB - BACKGROUND: The development of well-tolerated acid suppressant drugs has stimulated substantial growth in the number of trials assessing therapy options for gastro-oesophageal reflux disease (GERD). AIM: To develop consensus statements to inform clinical trial design in adult patients with GERD. METHODS: Draft statements were developed employing a systematic literature review. A modified Delphi process including three rounds of voting was used to reach consensus. Between voting, statements were revised based on feedback from the Working Group and additional literature reviews. The final vote was at a face-to face meeting that included discussion time. Voting was conducted using a six point scale. RESULTS: At the last vote, 93% of the final 102 statements achieved consensus (defined a priori as being supported by >or=75% of the votes). The Working Group strongly supported the development of validated patient-reported outcome instruments. Symptom assessments carried out by the investigator were considered unacceptable. There was agreement that exclusion from clinical trials should be minimized to improve generalizability, that prospective evaluation ideally requires electronic timed/dated methods and that endoscopists should be blinded to patient symptom status. CONCLUSIONS: Implementation of the consensus statements will improve the quality and comparability of trials, and make them compatible with regulatory requirements. PMID- 18384663 TI - Proton pump inhibitor therapy in gastro-oesophageal reflux disease decreases the oesophageal immune response but does not reduce the formation of DNA adducts. AB - BACKGROUND: Chronic oesophageal inflammation and related oxidative stress are important in the pathogenesis of erosive oesophagitis (EO) and its malignant progression. AIM: To study the effect of proton pump inhibitors (PPIs) on oesophageal cellular immune response and oxidative damage in EO patients. METHODS: Forty gastro-oesophageal reflux disease (GERD) patients [non-erosive reflux disease (NERD): 15, EO: 25] were included, after 7 days off antisuppressive drugs. EO patients were randomized to 20-mg rabeprazole once daily for either 4 or 8 weeks with baseline and follow-up endoscopy with distal oesophageal biopsies. T lymphocytes, macrophages and mast cells were quantified by immunohistochemistry. DNA adducts were measured by analysis of 8-oxo deoxyguanosine levels. RESULTS: Erosive oesophagitis patients had more T lymphocytes and CD8(+) T lymphocytes in squamous epithelium than NERD patients (P = 0.001, P = 0.002, respectively). Levels of DNA adducts between both groups were, however, not different (P = 0.99). Four- and eight-week rabeprazole treatment in EO patients resulted in a significant decrease in number of T lymphocytes and CD8(+) T lymphocytes (all P < 0.05). PPIs did not, however, affect levels of DNA adducts. CONCLUSIONS: Short-term PPI therapy in EO patients reduces the oesophageal cellular immune response, but does not change oxidative damage. PPI therapy may therefore not be effective in reducing the risk of oesophageal cancer in GERD patients. PMID- 18384664 TI - Review article: medication non-adherence in ulcerative colitis--strategies to improve adherence with mesalazine and other maintenance therapies. AB - BACKGROUND: Significant number of patients with ulcerative colitis (UC) fail to comply with treatment. AIMS: To review issues surrounding medication non adherence in inflammatory bowel disease (IBD), including the clinical and health service implications in the UK, and discuss strategies for optimizing medication adherence. METHODS: Articles cited were identified via a PubMed search, utilizing the words IBD, adherence, compliance, medication and UC. RESULTS: Medication non adherence is multifactorial involving factors other than dosing frequency. Male gender (OR: 2.06), new patient status (OR: 2.14), work and travel pressures (OR: 4.9) and shorter disease duration (OR: 2.1), among others are proven predictors of non-adherence in UC. These indicators can identify 'at-risk' patients and allow an individually tailored treatment approach to be introduced that optimizes medication adherence. A collaborative relationship between physician and patient is important; several strategies for improving adherence have been proven effective including open dialogue that takes into consideration the patient's health beliefs and concerns, providing educational (e.g. verbal/written information, self-management programmes) and behavioural interventions (e.g. calendar blister packs, cues/reminders). CONCLUSIONS: Educational and behavioural interventions tailored to individual patients can optimize medication adherence. Additional studies combining educational and behavioural interventions may provide further strategies for improving medication adherence rates in UC. PMID- 18384665 TI - Conditional knockdown of hMRS2 results in loss of mitochondrial Mg(2+) uptake and cell death. AB - The human gene MRS2L encodes a mitochondrial protein distantly related to CorA Mg(2+) transport proteins. Constitutive shRNA-mediated knockdown of hMRS2 in human HEK-293 cell line was found here to cause death. To further study its role in Mg(2+) transport, we have established stable cell lines with conditionally expressing shRNAs directed against hMRS2L. The cells expressing shRNA for several generations exhibited lower steady-state levels of free mitochondrial Mg(2+) ([Mg(2+)](m)) and reduced capacity of mitochondrial Mg(2+) uptake than control cells. Long-term expression of shRNAs resulted in loss of mitochondrial respiratory complex I, decreased mitochondrial membrane potential and cell death. We conclude that hMrs2 is the major transport protein for Mg (+) uptake into mitochondria and that expression of hMrs2 is essential for the maintenance of respiratory complex I and cell viability. PMID- 18384666 TI - An original SERPINA3 gene cluster: elucidation of genomic organization and gene expression in the Bos taurus 21q24 region. AB - BACKGROUND: The superfamily of serine proteinase inhibitors (serpins) is involved in numerous fundamental biological processes as inflammation, blood coagulation and apoptosis. Our interest is focused on the SERPINA3 sub-family. The major human plasma protease inhibitor, alpha1-antichymotrypsin, encoded by the SERPINA3 gene, is homologous to genes organized in clusters in several mammalian species. However, although there is a similar genic organization with a high degree of sequence conservation, the reactive-centre-loop domains, which are responsible for the protease specificity, show significant divergences. RESULTS: We provide additional information by analyzing the situation of SERPINA3 in the bovine genome. A cluster of eight genes and one pseudogene sharing a high degree of identity and the same structural organization was characterized. Bovine SERPINA3 genes were localized by radiation hybrid mapping on 21q24 and only spanned over 235 Kilobases. For all these genes, we propose a new nomenclature from SERPINA3-1 to SERPINA3-8. They share approximately 70% of identity with the human SERPINA3 homologue. In the cluster, we described an original sub-group of six members with an unexpected high degree of conservation for the reactive-centre-loop domain, suggesting a similar peptidase inhibitory pattern. Preliminary expression analyses of these bovSERPINA3s showed different tissue-specific patterns and diverse states of glycosylation and phosphorylation. Finally, in the context of phylogenetic analyses, we improved our knowledge on mammalian SERPINAs evolution. CONCLUSION: Our experimental results update data of the bovine genome sequencing, substantially increase the bovSERPINA3 sub-family and enrich the phylogenetic tree of serpins. We provide new opportunities for future investigations to approach the biological functions of this unusual subset of serine proteinase inhibitors. PMID- 18384667 TI - Construction of nested genetic core collections to optimize the exploitation of natural diversity in Vitis vinifera L. subsp. sativa. AB - BACKGROUND: The first high quality draft of the grape genome sequence has just been published. This is a critical step in accessing all the genes of this species and increases the chances of exploiting the natural genetic diversity through association genetics. However, our basic knowledge of the extent of allelic variation within the species is still not sufficient. Towards this goal, we constructed nested genetic core collections (G-cores) to capture the simple sequence repeat (SSR) diversity of the grape cultivated compartment (Vitis vinifera L. subsp. sativa) from the world's largest germplasm collection (Domaine de Vassal, INRA Herault, France), containing 2262 unique genotypes. RESULTS: Sub samples of 12, 24, 48 and 92 varieties of V. vinifera L. were selected based on their genotypes for 20 SSR markers using the M-strategy. They represent respectively 58%, 73%, 83% and 100% of total SSR diversity. The capture of allelic diversity was analyzed by sequencing three genes scattered throughout the genome on 233 individuals: 41 single nucleotide polymorphisms (SNPs) were identified using the G-92 core (one SNP for every 49 nucleotides) while only 25 were observed using a larger sample of 141 individuals selected on the basis of 50 morphological traits, thus demonstrating the reliability of the approach. CONCLUSION: The G-12 and G-24 core-collections displayed respectively 78% and 88% of the SNPs respectively, and are therefore of great interest for SNP discovery studies. Furthermore, the nested genetic core collections satisfactorily reflected the geographic and the genetic diversity of grape, which are also of great interest for the study of gene evolution in this species. PMID- 18384668 TI - Bridging the phenotypic gap: real-time assessment of mitochondrial function and metabolism of the nematode Caenorhabditis elegans. AB - BACKGROUND: The ATP levels of an organism are an important physiological parameter that is affected by genetic make up, ageing, stress and disease. RESULTS: We have generated luminescent C. elegans through ubiquitous, constitutive expression of firefly luciferase, widely used for in vitro ATP determination. We hypothesise that whole animal luminescence reflects its intracellular ATP levels in vivo. To test this, we characterised the bioluminescence response of C. elegans during sublethal exposure to, and recovery from azide, a treatment that inhibits mitochondrial respiration reversibly, and causes ATP depletion. Consistent with our expectations, in vivo luminescence decreased with increasing sublethal azide levels, and recovered fully when worms were removed from azide. Firefly luciferase expression levels, stability and activity did not influence the final luminescence. Bioluminescence also reflected the lowered activity of the electron transport chain achieved with RNA interference (RNAi) of genes encoding respiratory chain components. CONCLUSION: Results indicated that C. elegans luminescence reports on ATP levels in real time. For the first time, we are able to directly assess the metabolism of a whole, living, multicellular organism by determination of the relative ATP levels. This will enable genetic analysis based on a readily quantifiable metabolic phenotype and will provide novel insights into mechanisms of fitness and disease that are likely to be of relevance for other organisms, as well as the worm. PMID- 18384669 TI - Guidelines and mindlines: why do clinical staff over-diagnose malaria in Tanzania? A qualitative study. AB - BACKGROUND: Malaria over-diagnosis in Africa is widespread and costly both financially and in terms of morbidity and mortality from missed diagnoses. An understanding of the reasons behind malaria over-diagnosis is urgently needed to inform strategies for better targeting of antimalarials. METHODS: In an ethnographic study of clinical practice in two hospitals in Tanzania, 2,082 patient consultations with 34 clinicians were observed over a period of three months at each hospital. All clinicians were also interviewed individually as well as being observed during routine working activities with colleagues. Interviews with five tutors and 10 clinical officer students at a nearby clinical officer training college were subsequently conducted. RESULTS: Four, primarily social, spheres of influence on malaria over-diagnosis were identified. Firstly, the influence of initial training within a context where the importance of malaria is strongly promoted. Secondly, the influence of peers, conforming to perceived expectations from colleagues. Thirdly, pressure to conform with perceived patient preferences. Lastly, quality of diagnostic support, involving resource management, motivation and supervision. Rather than following national guidelines for the diagnosis of febrile illness, clinician behaviour appeared to follow 'mindlines': shared rationales constructed from these different spheres of influence. Three mindlines were identified in this setting: malaria is easier to diagnose than alternative diseases; malaria is a more acceptable diagnosis; and missing malaria is indefensible. These mindlines were apparent during the training stages as well as throughout clinical careers. CONCLUSION: Clinicians were found to follow mindlines as well as or rather than guidelines, which incorporated multiple social influences operating in the immediate and the wider context of decision making. Interventions to move mindlines closer to guidelines need to take the variety of social influences into account. PMID- 18384670 TI - Association between serum levels of C-reactive protein and personality traits in women. AB - BACKGROUND: While low-grade inflammation has consistently been observed in subjects with depression, studies on the possible relationship between inflammation and other aspects of brain function are as yet sparse. In this study, we aimed to investigate the possible association between serum levels of the inflammation marker C-reactive protein (CRP) and personality traits. METHODS: In this study, serum levels of high-sensitivity CRP were determined by ELISA in a population of 270 42-year-old women recruited from the population registry who had been assessed using the Temperament and Character Inventory. Self-reported previous or ongoing depression was also recorded. Unpaired two-tailed t-tests were used for comparison between two groups and correlations were evaluated by the calculation of Pearson's r-coefficient. RESULTS: The temperament trait harm avoidance was positively (r = 0.227, p < 0.05) and the character trait self directedness was negatively (r = -0.261, p < 0.01) associated with serum levels of CRP (p-values corrected for multiple comparisons). The correlations between the personality traits and CRP were observed also after exclusion of subjects reporting ongoing depression (n = 26). Whereas women reporting ongoing depression showed significantly increased levels of CRP as compared to non-depressed women (n = 155), women reporting a history of depression displayed no significant difference in CRP levels as compared to women that reported that they had never been depressed. CONCLUSION: Serum levels of CRP in women was found to be associated with the personality traits harm avoidance and self-directedness. In addition, moderately elevated levels may be a state dependent marker of depression. PMID- 18384671 TI - Weak correlation between sequence conservation in promoter regions and in protein coding regions of human-mouse orthologous gene pairs. AB - BACKGROUND: Interspecies sequence comparison is a powerful tool to extract functional or evolutionary information from the genomes of organisms. A number of studies have compared protein sequences or promoter sequences between mammals, which provided many insights into genomics. However, the correlation between protein conservation and promoter conservation remains controversial. RESULTS: We examined promoter conservation as well as protein conservation for 6,901 human and mouse orthologous genes, and observed a very weak correlation between them. We further investigated their relationship by decomposing it based on functional categories, and identified categories with significant tendencies. Remarkably, the 'ribosome' category showed significantly low promoter conservation, despite its high protein conservation, and the 'extracellular matrix' category showed significantly high promoter conservation, in spite of its low protein conservation. CONCLUSION: Our results show the relation of gene function to protein conservation and promoter conservation, and revealed that there seem to be nonparallel components between protein and promoter sequence evolution. PMID- 18384672 TI - Risk factors for the development of nosocomial pneumonia and mortality on intensive care units: application of competing risks models. AB - INTRODUCTION: Pneumonia is a very common nosocomial infection in intensive care units (ICUs). Many studies have investigated risk factors for the development of infection and its consequences. However, the evaluation in most of theses studies disregards the fact that there are additional competing events, such as discharge or death. METHODS: A prospective cohort study was conducted over 18 months in five intensive care units at one university hospital. All patients that were admitted for at least 2 days were included, and surveillance of nosocomial pneumonia was conducted. Various potential risk factors (baseline- and time dependent) were evaluated in two competing risks models: the acquisition of nosocomial pneumonia and discharge (dead or alive; model 1) and for the risk of death in the ICU and discharge alive (model 2). RESULTS: Patients from 1,876 admissions were included. A total of 158 patients developed nosocomial pneumonia. The main risk factors for nosocomial pneumonia in the multivariate analysis in model 1 were: elective surgery (cause-specific hazard ratio = 1.95; 95% CI 1.33 to 2.85) or emergency surgery (1.59; 95% CI 1.10 to 2.28) prior to ICU admission, usage of a nasogastric tube (3.04; 95% CI 1.25 to 7.37) and mechanical ventilation (5.90; 95% CI 2.47 to 14.09). Nosocomial pneumonia prolonged the length of ICU stay but was not directly associated with a fatal outcome (p = 0.55). CONCLUSION: More studies using competing risk models, which provide more accurate data compared to naive survival curves or logistic models, should be carried out to verify the impact of risk factors and patient characteristics for the acquisition of nosocomial infections and infection-associated mortality. PMID- 18384673 TI - R-spondin1 and FOXL2 act into two distinct cellular types during goat ovarian differentiation. AB - BACKGROUND: Up to now, two loci have been involved in XX sex-reversal in mammals following loss-of-function mutations, PIS (Polled Intersex Syndrome) in goats and R-spondin1 (RSPO1) in humans. Here, we analyze the possible interaction between these two factors during goat gonad development. Furthermore, since functional redundancy between different R-spondins may influence gonad development, we also studied the expression patterns of RSPO2, 3 and 4. RESULTS: Similarly to the mouse, RSPO1 shows a sex-dimorphic expression pattern during goat gonad development with higher levels in the ovaries. Interestingly, the PIS mutation does not seem to influence its level of expression. Moreover, using an RSPO1 specific antibody, the RSPO1 protein was localized in the cortical area of early differentiating ovaries (36 and 40 dpc). This cortical area contains the majority of germ cell that are surrounded by FOXL2 negative somatic cells. At latter stages (50 and 60 dpc) RSPO1 protein remains specifically localized on the germ cell membranes. Interestingly, a time-specific relocation of RSPO1 on the germ cell membrane was noticed, moving from a uniform distribution at 40 dpc to a punctuated staining before and during meiosis (50 and 60 dpc respectively). Interestingly, also RSPO2 and RSPO4 show a sex-dimorphic expression pattern with higher levels in the ovaries. Although RSPO4 was found to be faintly and belatedly expressed, the expression of RSPO2 increases at the crucial 36 dpc stage, as does that of FOXL2. Importantly, RSPO2 expression appears dramatically decreased in XX PIS-/- gonads at all three tested stages (36, 40 and 50 dpc). CONCLUSION: During goat ovarian development, the pattern of expression of RSPO1 is in agreement with its possible anti-testis function but is not influenced by the PIS mutation. Moreover, our data suggest that RSPO1 may be associated with germ cell development and meiosis. Interestingly, another RSPO gene, RSPO2 shows a sex-dimorphic pattern of expression that is dramatically influenced by the PIS mutation. PMID- 18384674 TI - Food-intake-regulating-neuropeptides are expressed and regulated through pregnancy and following food restriction in rat placenta. AB - BACKGROUND: Neuropeptide Y (NPY), agouti related peptide (AgRP), cocaine and amphetamine-regulated transcript (CART) and melanocortins, the products of the proopiomelanocortin (POMC), are hypothalamic peptides involved in feeding regulation and energy homeostasis. Recent evidence has demonstrated their expression in rat and human placenta. METHODS: In the current study, we have investigated the expression of those neuropeptides in the rat placenta by real time PCR using a model of maternal food restriction. RESULTS: Our results showed that placental-derived neuropeptides were regulated through pregnancy and following food restriction. CONCLUSION: These data could indicate that placental derived neuropeptides represent a local regulatory circuit that may fine-tune control of energy balance during pregnancy. PMID- 18384675 TI - Can an EASYcare based dementia training programme improve diagnostic assessment and management of dementia by general practitioners and primary care nurses? The design of a randomised controlled trial. AB - BACKGROUND: Early diagnosis of dementia benefits both patient and caregiver. Nevertheless, dementia in primary care is currently under-diagnosed. Some educational interventions developed to improve dementia diagnosis and management were successful in increasing the number of dementia diagnoses and in changing attitudes and knowledge of health care staff. However, none of these interventions focussed on collaboration between GPs and nurses in dementia care. We developed an EASYcare-based Dementia Training Program (DTP) aimed at stimulating collaboration in dementia primary care. We expect this program to increase the number of cognitive assessments and dementia diagnoses and to improve attitudes and knowledge of GPs and nurses. METHODS: The DTP is a complex educational intervention that consists of two workshops, a coaching program, access to an internet forum, and a Computerized Clinical Decision Support System on dementia diagnostics. One hundred duos of GPs and nurses will be recruited, from which 2/3 will be allocated to the intervention group and 1/3 to the control group. The effects of implementation of the DTP will be studied in a cluster randomised controlled trial. Primary outcomes will be the number of cognitive assessments and dementia diagnoses in a period of 9 months following workshop participation. Secondary outcomes are measured on GP and nurse level: adherence to national guidelines for dementia, attitude, confidence and knowledge regarding dementia diagnosis and management; on patient level: number of emergency calls, visits and consultations and patient satisfaction; and on caregiver level: informal caregiver burden and satisfaction. Data will be collected from GPs' electronic medical records, self-registration forms and questionnaires. Statistical analysis will be performed using the MANOVA-method. Also, exploratory analyses will be performed, in order to gain insight into barriers and facilitators for implementation and the possible causal relations between the rate of success of the intervention components and the outcomes. DISCUSSION: We developed multifaceted dementia training programme. Novelties in this programme are the training in fixed collaborative duos and the inclusion of an individual coaching program. The intervention is designed according to international guidelines and educational standards. Exploratory analysis will reveal its successful elements. Selection bias and contamination may be threats to the reliability of future results of this trial. Nevertheless, the results of this trial may provide useful information for policy makers and developers of continuing medical education. TRIAL REGISTRATION: ClinicalTrials.gov ID NCT00459784. PMID- 18384676 TI - Ornithine decarboxylase antizyme finder (OAF): fast and reliable detection of antizymes with frameshifts in mRNAs. AB - BACKGROUND: Ornithine decarboxylase antizymes are proteins which negatively regulate cellular polyamine levels via their affects on polyamine synthesis and cellular uptake. In virtually all organisms from yeast to mammals, antizymes are encoded by two partially overlapping open reading frames (ORFs). A +1 frameshift between frames is required for the synthesis of antizyme. Ribosomes change translation phase at the end of the first ORF in response to stimulatory signals embedded in mRNA. Since standard sequence analysis pipelines are currently unable to recognise sites of programmed ribosomal frameshifting, proper detection of full length antizyme coding sequences (CDS) requires conscientious manual evaluation by a human expert. The rapid growth of sequence information demands less laborious and more cost efficient solutions for this problem. This manuscript describes a rapid and accurate computer tool for antizyme CDS detection that requires minimal human involvement. RESULTS: We have developed a computer tool, OAF (ODC antizyme finder) for identifying antizyme encoding sequences in spliced or intronless nucleic acid sequenes. OAF utilizes a combination of profile hidden Markov models (HMM) built separately for the products of each open reading frame constituting the entire antizyme coding sequence. Profile HMMs are based on a set of 218 manually assembled antizyme sequences. To distinguish between antizyme paralogs and orthologs from major phyla, antizyme sequences were clustered into twelve groups and specific combinations of profile HMMs were designed for each group. OAF has been tested on the current version of dbEST, where it identified over six thousand Expressed Sequence Tags (EST) sequences encoding antizyme proteins (over two thousand antizyme CDS in these ESTs are non redundant). CONCLUSION: OAF performs well on raw EST sequences and mRNA sequences derived from genomic annotations. OAF will be used for the future updates of the RECODE database. OAF can also be useful for identifying novel antizyme sequences when run with relaxed parameters. It is anticipated that OAF will be used for EST and genome annotation purposes. OAF outputs sequence annotations in fasta, genbank flat file or XML format. The OAF web interface and the source code are freely available at http://recode.ucc.ie/oaf/ and at a mirror site http://recode.genetics.utah.edu/oaf/. PMID- 18384677 TI - Effect of early onset otitis media on brainstem and cortical auditory processing. AB - BACKGROUND: Otitis media (OM) leads to significant reduction in the hearing sensitivity. The reduced auditory input, if in the early years of life when the auditory neural system is still maturing, may adversely influence the structural as well as functional development of the system. Past research has reported abnormalities in both the structure and function of brainstem nuclei following auditory deprivation, but, it has not necessarily focused on children who had OM in their first year of life. It can also be said that if auditory processing is affected at the brainstem level because of early onset OM (reduced auditory input in the crucial periods of neural development), then, it may be said that auditory processing is also affected at the cortical level because it receives distorted input from the brainstem. Therefore, the purpose of this study was to document the effects of early onset OM on auditory processing, if any, at the brainstem as well as at cortical levels. A related purpose of the study was to investigate the persistence of the effects of early onset OM, if any, on auditory processing. METHODS: A cross sectional approach and a standard group comparison design was used in the study. Thirty children, who had OM between 6 and 12 months of age and who were in the age range of 3.1 - 5.6 years participated in the study. Children with OM were divided into 3 groups based on their age. Click evoked auditory brainstem responses (ABRs) and late latency responses (LLRs) were recorded from these children, and the responses were compared with those from age and gender matched normal children without any history of OM. The data from the 2 groups was statistically analyzed through independent t test. Pearson's Product Moment correlation was computed to examine the relationship between results of ABR and LLR in children with early onset OM. RESULTS: The mean central conduction time was significantly increased and the mean amplitude of wave I and III of ABRs was significantly reduced in children with early onset OM compared to normal children. Also, the latency of all LLR waves was significantly less in children with early onset OM than in normal children. However, significant differences in mean values of either ABR or LLR (latencies or interwave intervals as the case may be) were observed only in 3-year old children. There was a significant, but negative association between central conduction time and latency of LLRs. CONCLUSION: OM in the first year of life leads to negative effects on brainstem signal processing even if it has occurred only for a short duration (maximum of 3 months). In such a situation, auditory cortical structures probably show compensatory changes through central gain to offset the prolonged central conduction time. Although the results of the present study showed that the negative effects of early onset OM (occurring in the first year of life) on auditory processing disappeared by the time the children were 4.1 years, there is need for longitudinal studies on this to confirm the findings. PMID- 18384678 TI - No temporal trends in the prevalence of atypical scrapie in British sheep, 2002 2006. AB - BACKGROUND: So-called atypical scrapie was first identified in Great Britain (GB) in 2002 following the introduction of wide-scale scrapie surveillance. In particular, abattoir and fallen stock surveys have been carried out in GB since 2002, with a total of 147 atypical positives identified by the end of 2006. The results of these surveys provide data with which to assess temporal trends in the prevalence of atypical scrapie in sheep in Great Britain between 2002 and 2006. RESULTS: Using the results of abattoir and fallen stock surveys, the prevalence of atypical scrapie (percentage of samples positive) was estimated. The prevalence in the abattoir and fallen stock surveys, for all years combined, was 0.09% (95% confidence interval (CI): 0.08%-0.11%) and 0.07% (95% CI: 0.05% 0.11%), respectively. There were no significant temporal trends in either survey. Comparing the surveys' results, there were no significant differences in annual prevalence or the prevalence within PrP genotypes. For the abattoir survey, the PrP genotype with the highest prevalence was AHQ/AHQ, which was significantly higher than all other genotypes, except ARR/AHQ, AHQ/ARH and ARH/ARQ. CONCLUSION: The estimated prevalence of atypical scrapie was similar in both the abattoir and fallen stock surveys. Our results indicate there was no significant temporal trend in prevalence, adding to evidence that this atypical form of scrapie may be a sporadic condition or, if it is infectious, that the force of infection is very low. PMID- 18384679 TI - Numbers needed to treat calculated from responder rates give a better indication of efficacy in osteoarthritis trials than mean pain scores. AB - INTRODUCTION: Osteoarthritis trials usually report average changes in visual analogue scale (VAS) pain, and examine the difference between treatment and placebo. We investigated whether dichotomous responder analysis provides a more informative interpretation of drug efficacy. METHODS: Merck supplied the number of patients who, by 6 weeks, had achieved pain relief compared with a baseline of 0% or more, 10% or more, 20% or more, and so on at equal intervals up to 90% or more. These different levels of pain relief were used to distinguish different definitions of responders, for example at least 50% pain relief from baseline. Numbers and percentages of patients achieving each level were identified. Information was sought from a dose-response trial over 6 weeks in osteoarthritis using placebo and using etoricoxib at 5, 10, 30 and 60 mg daily. RESULTS: With placebo, the proportions of patients achieving at least 20%, 50% and 70% pain relief over baseline at 6 weeks were 30%, 11% and 2%. With 60 mg etoricoxib the equivalent percentages were 74%, 49% and 29%. The numbers needed to treat for 30 mg and 60 mg etoricoxib to produce at least 50% pain relief at 6 weeks compared with placebo were 4.2 (95% confidence interval 3.8 to 8.6) and 2.6 (2.0 to 3.9), respectively. Levels of pain relief of 50% and above discriminated best between different doses of etoricoxib. CONCLUSION: Responder analysis seemed to be more sensitive than examination of average changes in VAS pain scores. Validation would require calculations to be performed on a set of trials using individual patient data not available in publications. PMID- 18384680 TI - Testing for hereditary thrombophilia: a retrospective analysis of testing referred to a national laboratory. AB - BACKGROUND: Predisposition to venous thrombosis may be assessed through testing for defects and/or deficiencies of a number of hereditary factors. There is potential for confusion about which of these tests are appropriate in which settings. At least one set of recommendations has been published to guide such testing, but it is unclear how widely these have been disseminated. METHODS: We performed a retrospective analysis of laboratory orders and results at a national referral laboratory to gain insight into physicians' ordering practices, specifically comparing them against the ordering practices recommended by a 2002 College of American Pathologists (CAP) consensus conference on thrombophilia testing. Measurements included absolute and relative ordering volumes and positivity rates from approximately 200,000 thrombophilia tests performed from September 2005 through August 2006 at a national reference laboratory. Quality control data were used to estimate the proportion of samples that may have been affected by anticoagulant therapy. A sample of ordering laboratories was surveyed in order to assess potential measurement bias. RESULTS: Total antigen assays for protein C, protein S and antithrombin were ordered almost as frequently as functional assays for these analytes. The DNA test for factor V Leiden was ordered much more often than the corresponding functional assay. In addition, relative positivity rates coupled with elevations in prothrombin time (PT) in many of these patients suggest that these tests are often ordered in the setting of oral anticoagulant therapy. CONCLUSION: In this real-world setting, testing for inherited thrombophilia is frequently at odds with the recommendations of the CAP consensus conference. There is a need for wider dissemination of concise thrombophilia testing guidelines. PMID- 18384681 TI - Surgical impact on brain tumor invasion: a physical perspective. AB - It is conventional strategy to treat highly malignant brain tumors initially with cytoreductive surgery followed by adjuvant radio- and chemotherapy. However, in spite of all such efforts, the patients' prognosis remains dismal since residual glioma cells continue to infiltrate adjacent parenchyma and the tumors almost always recur. On the basis of a simple biomechanical conjecture that we have introduced previously, we argue here that by affecting the 'volume-pressure' relationship and minimizing surface tension of the remaining tumor cells, gross total resection may have an inductive effect on the invasiveness of the tumor cells left behind. Potential implications for treatment strategies are discussed. PMID- 18384682 TI - Physicians' perceptions of an electronic health record-based clinical trial alert approach to subject recruitment: a survey. AB - BACKGROUND: Physician participation in clinical research recruitment efforts is critical to many studies' success, but it is often limited. Use of an Electronic Health Record (EHR)-based, point-of-care Clinical Trial Alert (CTA) approach has led to significant increases in physician-generated recruitment and holds promise for wider benefit. However, little is known about physicians' decision-making regarding recruitment in EHR-equipped settings or the use of such EHR-based approaches. We sought to assess physicians' perceptions about recruitment in general and using the CTA approach in particular. METHODS: We developed and delivered a Web-based survey consisting of 15 multiple-choice and free-text questions. Participants included the 114 physician subjects (10 endocrinologists and 104 general internists) who were exposed to CTAs during our preceding 4-month intervention study. Response data were descriptively analyzed, and key findings were compared between groups using appropriate statistical tests. RESULTS: Sixty nine physicians (61%) responded during the 10-week survey period. Respondents and non-respondents did not differ significantly. Twenty-seven percent of respondents felt very comfortable recruiting patients to trials in general, and 77% appreciated being reminded about a trial via a CTA. Only 11% percent felt the CTA was difficult to use, and 27% felt it was more than somewhat intrusive. Among those who ignored all CTAs, 37% cited a lack of time, 28% knowledge of the patient's ineligibility, and 13% limited knowledge about the trial as their most common reason. Thirty-eight percent wanted more information about the trial presented in the CTA, and 73% were interested in seeing CTAs for future trials. Comments and suggestions were submitted by 33% of respondents and included suggestions for improvement of the CTA approach. CONCLUSION: Most physicians were comfortable recruiting patients for clinical trials at the point-of-care, found the EHR-based CTA approach useful and would like to see it used in the future. These findings provide insight into the perceived utility of this EHR-based approach to subject recruitment, suggest ways it might be improved, and add to the limited body of knowledge regarding physicians' attitudes toward clinical trial recruitment in EHR-equipped settings. PMID- 18384683 TI - Coliform pyosalpinx as a rare complication of appendicectomy: a case report and review of the literature on best practice. AB - INTRODUCTION: Coliform pyosalpinx is a rare entity. We report a case that occurred three months after appendicectomy for gangrenous appendicitis. There follows a literature review on best practice for the treatment of pyosalpinx. CASE PRESENTATION: A seventeen year old girl presented with an acute abdomen three months after an appendicectomy for gangrenous appendicitis. Intraoperative findings were bilateral pyosalpinx treated by aspiration, saline and Betadine irrigation and intravenous antibiotics. CONCLUSION: Microbiological analysis of the pus revealed Escherichia coli and anaerobes. Chlamydia and Candida were not isolated. This is the first known reported case of Coliform Pyosalpinx following appendicectomy. The best treatment does not necessarily involve salpingectomy especially in women of reproductive age where fertility may become compromised. PMID- 18384684 TI - Prevalence and correlates of being bullied among in-school adolescents in Beijing: results from the 2003 Beijing Global School-Based Health Survey. AB - BACKGROUND: Bullying has public health importance. It has been reported that both the victims and perpetrators of bullying are more likely to have suicidal ideation and other suicidal behaviours. Moreover, bullying can be a precursor for school violence and can contribute to poor academic performance. The purpose of the study was to raise awareness on the subject in China. We, therefore conducted an analysis of secondary data to determine the prevalence and correlates of having been bullied among in-school adolescents. METHODS: The data was taken from the Beijing Global School-Based Health Survey conducted in 2003. A weighted analysis to reduce bias due to differing patterns of non-response was conducted using statistical software (SPSS version 14.0). We conducted a backward logistic regression analysis to determine independent predictors for being bullied. RESULTS: Out of a total of 2,348 in-school adolescents who participated in the survey, 20% (23% males, and 17% females) reported having been bullied. Risk factors for having been bullied were loneliness (adjusted odds ratio (AOR) = 1.44; 95% confidence interval (CI) 1.42-1.45), being worried (AOR = 1.30; 95% CI 1.29-1.32), being sad or having feelings of hopelessness (AOR = 1.21; 95% CI 1.19 1.22), smoking cigarettes (AOR = 1.09; 95% CI 1.08-1.11), drinking alcohol (AOR = 1.31; 95% CI 1.29-1.32), and being truant (AOR = 1.24; 95% CI 1.22-1.27). Meanwhile protective factors were having close friends (AOR = 0.84; 95% CI 0.83 0.86), receiving parental supervision (AOR = 0.80; 95% CI 0.80-0.81), and ever been drunk (AOR = 0.86; 95% CI 0.84-0.87). CONCLUSION: We believe the results of this study will raise awareness among school health practitioners and administrators, paediatric psychiatrists and psychologists on the prevalence and correlates of bullying among adolescents in Beijing, China. PMID- 18384685 TI - Recovery of a Burkholderia thailandensis-like isolate from an Australian water source. AB - BACKGROUND: Burkholderia thailandensis, a close relative of Burkholderia pseudomallei, has previously been reported only from Southeast Asia and North America. It is biochemically differentiated from B. pseudomallei by the ability to utilize arabinose. During the course of environmental sampling for B. pseudomallei in the Northern Territory of Australia, an isolate, MSMB 43, was recovered that is arabinose positive. RESULTS: Genetic analysis using 16S rDNA sequencing and DNA/DNA hybridization indicates that MSMB 43 is most similar to B. thailandensis although multi-locus sequence typing indicates that this isolate is divergent from both B. pseudomallei and other described B. thailandensis. CONCLUSION: We report the isolation and initial characterization of strain MSMB 43, which is a B. thailandensis-like isolate recovered in Australia. PMID- 18384686 TI - Spatial separation and bidirectional trafficking of proteins using a multi functional reporter. AB - BACKGROUND: The ability to specifically label proteins within living cells can provide information about their dynamics and function. To study a membrane protein, we fused a multi-functional reporter protein, HaloTag, to the extracellular domain of a truncated integrin. RESULTS: Using the HaloTag technology, we could study the localization, trafficking and processing of an integrin-HaloTag fusion, which we showed had cellular dynamics consistent with native integrins. By labeling live cells with different fluorescent impermeable and permeable ligands, we showed spatial separation of plasma membrane and internal pools of the integrin-HaloTag fusion, and followed these protein pools over time to study bi-directional trafficking. In addition to combining the HaloTag reporter protein with different fluorophores, we also employed an affinity tag to achieve cell capture. CONCLUSION: The HaloTag technology was used successfully to study expression, trafficking, spatial separation and real-time translocation of an integrin-HaloTag fusion, thereby demonstrating that this technology can be a powerful tool to investigate membrane protein biology in live cells. PMID- 18384687 TI - Pro-protein convertases control the maturation and processing of the iron regulatory protein, RGMc/hemojuvelin. AB - BACKGROUND: Repulsive guidance molecule c (RGMc or hemojuvelin), a glycosylphosphatidylinositol-linked glycoprotein expressed in liver and striated muscle, plays a central role in systemic iron balance. Inactivating mutations in the RGMc gene cause juvenile hemochromatosis (JH), a rapidly progressing iron storage disorder with severe systemic manifestations. RGMc undergoes complex biosynthetic steps leading to membrane-bound and soluble forms of the protein, including both 50 and 40 kDa single-chain species. RESULTS: We now show that pro protein convertases (PC) are responsible for conversion of 50 kDa RGMc to a 40 kDa protein with a truncated COOH-terminus. Unlike related molecules RGMa and RGMb, RGMc encodes a conserved PC recognition and cleavage site, and JH associated RGMc frame-shift mutants undergo COOH-terminal cleavage only if this site is present. A cell-impermeable peptide PC inhibitor blocks the appearance of 40 kDa RGMc in extra-cellular fluid, as does an engineered mutation in the conserved PC recognition sequence, while the PC furin cleaves 50 kDa RGMc in vitro into a 40 kDa molecule with an intact NH2-terminus. Iron loading reduces release of RGMc from the cell membrane, and diminishes accumulation of the 40 kDa species in cell culture medium. CONCLUSION: Our results define a role for PCs in the maturation of RGMc that may have implications for the physiological actions of this critical iron-regulatory protein. PMID- 18384688 TI - The assessment of angiogenesis and fibroblastic stromagenesis in hyperplastic and pre-invasive breast lesions. AB - BACKGROUND: To investigate the changes of the neoplastic microenvironment during the different morphological alterations of hyperplastic and pre-invasive breast lesions. METHODS: 78 in situ ductal carcinomas of all degrees of differentiation, 22 atypical ductal hyperplasias, 25 in situ lobular carcinomas, 18 atypical lobular hyperplasias, 32 ductal epithelial hyperplasias of usual type and 8 flat atypias were immunohistochemically investigated for the expression of vascular endothelial growth factor (VEGF), smooth muscle actin (SMA) and CD34, while microvessel density (MVD) was counted using the anti-CD31 antibody. RESULTS: VEGF expression was strongly correlated with MVD in all hyperplastic and pre-invasive breast lesions (p < 0.05). Stromagenesis, as characterized by an increase in SMA and a decrease in CD34 positive myofibroblasts was observed mostly around ducts harboring high grade in situ carcinoma and to a lesser extent around moderately differentiated DCIS. In these two groups of in situ carcinomas, a positive correlation between MVD and SMA (p < 0.05) was observed. On the contrary, CD34 was found to be inversely related to MVD (p < 0.05). No statistically significant changes of the stromal fibroblasts were observed in low grade DCIS neither in any of the other lesions under investigation as compared to normal mammary intra- and interlobular stroma. CONCLUSION: Angiogenesis is observed before any significant fibroblastic stromagenesis in pre-invasive breast lesions. A composite phenotype characterized by VEGF positive epithelial cells and SMA positive/CD34 negative stromal cells, is identified mostly in intermediate and high grade DCIS. These findings might imply for new therapeutic strategies using both anti-angiogenic factors and factors selectively targeting tumor stroma in order to prevent the progression of DCIS to invasive carcinoma. PMID- 18384689 TI - Evolution of pharmacologic specificity in the pregnane X receptor. AB - BACKGROUND: The pregnane X receptor (PXR) shows the highest degree of cross species sequence diversity of any of the vertebrate nuclear hormone receptors. In this study, we determined the pharmacophores for activation of human, mouse, rat, rabbit, chicken, and zebrafish PXRs, using a common set of sixteen ligands. In addition, we compared in detail the selectivity of human and zebrafish PXRs for steroidal compounds and xenobiotics. The ligand activation properties of the Western clawed frog (Xenopus tropicalis) PXR and that of a putative vitamin D receptor (VDR)/PXR cloned in this study from the chordate invertebrate sea squirt (Ciona intestinalis) were also investigated. RESULTS: Using a common set of ligands, human, mouse, and rat PXRs share structurally similar pharmacophores consisting of hydrophobic features and widely spaced excluded volumes indicative of large binding pockets. Zebrafish PXR has the most sterically constrained pharmacophore of the PXRs analyzed, suggesting a smaller ligand-binding pocket than the other PXRs. Chicken PXR possesses a symmetrical pharmacophore with four hydrophobes, a hydrogen bond acceptor, as well as excluded volumes. Comparison of human and zebrafish PXRs for a wide range of possible activators revealed that zebrafish PXR is activated by a subset of human PXR agonists. The Ciona VDR/PXR showed low sequence identity to vertebrate VDRs and PXRs in the ligand-binding domain and was preferentially activated by planar xenobiotics including 6 formylindolo-[3,2-b]carbazole. Lastly, the Western clawed frog (Xenopus tropicalis) PXR was insensitive to vitamins and steroidal compounds and was activated only by benzoates. CONCLUSION: In contrast to other nuclear hormone receptors, PXRs show significant differences in ligand specificity across species. By pharmacophore analysis, certain PXRs share similar features such as human, mouse, and rat PXRs, suggesting overlap of function and perhaps common evolutionary forces. The Western clawed frog PXR, like that described for African clawed frog PXRs, has diverged considerably in ligand selectivity from fish, bird, and mammalian PXRs. PMID- 18384690 TI - Failure to replicate an association of SNPs in the oxidized LDL receptor gene (OLR1) with CAD. AB - BACKGROUND: The lectin-like oxidized LDL receptor LOX-1 (encoded by OLR1) is believed to play a key role in atherogenesis and some reports suggest an association of OLR1 polymorphisms with myocardial infarction (MI). We tested whether single nucleotide polymorphisms (SNPs) in OLR1 are associated with clinically significant CAD in the Atherosclerotic Disease, VAscular FuNction, & Geneti C Epidemiology (ADVANCE) study. METHODS: ADVANCE is a population-based case-control study of subjects receiving care within Kaiser Permanente of Northern California including a subset of participants of the Coronary Artery Risk Development in Young Adults (CARDIA) study. We first resequenced the promoter, exonic, and splice site regions of OLR1 and then genotyped four single nucleotide polymorphisms (SNPs), including a non-synonymous SNP (rs11053646, Lys167Asn) as well as an intronic SNP (rs3736232) previously associated with CAD. RESULTS: In 1,809 cases with clinical CAD and 1,734 controls, the minor allele of the coding SNP was nominally associated with a lower odds ratio (OR) of CAD across all ethnic groups studied (minimally adjusted OR 0.8, P = 0.007; fully adjusted OR 0.8, P = 0.01). The intronic SNP was nominally associated with an increased risk of CAD (minimally adjusted OR 1.12, p = 0.03; fully adjusted OR 1.13, P = 0.03). However, these associations were not replicated in over 13,200 individuals (including 1,470 cases) in the Atherosclerosis Risk in Communities (ARIC) study. CONCLUSION: Our results do not support the presence of an association between selected common SNPs in OLR1 and the risk of clinical CAD. PMID- 18384691 TI - Nanopolymers improve delivery of exon skipping oligonucleotides and concomitant dystrophin expression in skeletal muscle of mdx mice. AB - BACKGROUND: Exon skipping oligonucleotides (ESOs) of 2'O-Methyl (2'OMe) and morpholino chemistry have been shown to restore dystrophin expression in muscle fibers from the mdx mouse, and are currently being tested in phase I clinical trials for Duchenne Muscular Dystrophy (DMD). However, ESOs remain limited in their effectiveness because of an inadequate delivery profile. Synthetic cationic copolymers of poly(ethylene imine) (PEI) and poly(ethylene glycol) (PEG) are regarded as effective agents for enhanced delivery of nucleic acids in various applications. RESULTS: We examined whether PEG-PEI copolymers can facilitate ESO mediated dystrophin expression after intramuscular injections into tibialis anterior (TA) muscles of mdx mice. We utilized a set of PEG-PEI copolymers containing 2 kDa PEI and either 550 Da or 5 kDa PEG, both of which bind 2'OMe ESOs with high affinity and form stable nanoparticulates with a relatively low surface charge. Three weekly intramuscular injections of 5 microg of ESO complexed with PEI2K-PEG550 copolymers resulted in about 500 dystrophin-positive fibers and about 12% of normal levels of dystrophin expression at 3 weeks after the initial injection, which is significantly greater than for injections of ESO alone, which are known to be almost completely ineffective. In an effort to enhance biocompatibility and cellular uptake, the PEI2K-PEG550 and PEI2K-PEG5K copolymers were functionalized by covalent conjugation with nanogold (NG) or adsorbtion of colloidal gold (CG), respectively. Surprisingly, using the same injection and dosing regimen, we found no significant difference in dystrophin expression by Western blot between the NG-PEI2K-PEG550, CG-PEI2K-PEG5K, and non functionalized PEI2K-PEG550 copolymers. Dose-response experiments using the CG PEI2K-PEG5K copolymer with total ESO ranging from 3-60 microg yielded a maximum of about 15% dystrophin expression. Further improvements in dystrophin expression up to 20% of normal levels were found at 6 weeks after 10 twice-weekly injections of the NG-PEI2K-PEG550 copolymer complexed with 5 microg of ESO per injection. This injection and dosing regimen showed over 1000 dystrophin-positive fibers. H&E staining of all treated muscle groups revealed no overt signs of cytotoxicity. CONCLUSION: We conclude that PEGylated PEI2K copolymers are efficient carriers for local delivery of 2'OMe ESOs and warrant further development as potential therapeutics for treatment of DMD. PMID- 18384692 TI - The pituitary tumor transforming gene 1 (PTTG-1): an immunological target for multiple myeloma. AB - BACKGROUND: Multiple Myeloma is a cancer of B plasma cells, which produce non specific antibodies and proliferate uncontrolled. Due to the potential relapse and non-specificity of current treatments, immunotherapy promises to be more specific and may induce long-term immunity in patients. The pituitary tumor transforming gene 1 (PTTG-1) has been shown to be a novel oncogene, expressed in the testis, thymus, colon, lung and placenta (undetectable in most other tissues). Furthermore, it is over expressed in many tumors such as the pituitary adenoma, breast, gastrointestinal cancers, leukemia, lymphoma, and lung cancer and it seems to be associated with tumorigenesis, angiogenesis and cancer progression. The purpose was to investigate the presence/rate of expression of PTTG-1 in multiple myeloma patients. METHODS: We analyzed the PTTG-1 expression at the transcriptional and the protein level, by PCR, immunocytochemical methods, Dot-blot and ELISA performed on patient's sera in 19 multiple myeloma patients, 6 different multiple myeloma cell lines and in normal human tissue. RESULTS: We did not find PTTG-1 presence in the normal human tissue panel, but PTTG-1 mRNA was detectable in 12 of the 19 patients, giving evidence of a 63% rate of expression (data confirmed by ELISA). Four of the 6 investigated cell lines (66.6%) were positive for PTTG-1. Investigations of protein expression gave evidence of 26.3% cytoplasmic expression and 16% surface expression in the plasma cells of multiple myeloma patients. Protein presence was also confirmed by Dot-blot in both cell lines and patients. CONCLUSION: We established PTTG-1's presence at both the transcriptional and protein levels. These data suggest that PTTG-1 is aberrantly expressed in multiple myeloma plasma cells, is highly immunogenic and is a suitable target for immunotherapy of multiple myeloma. PMID- 18384693 TI - Development of patatin knockdown potato tubers using RNA interference (RNAi) technology, for the production of human-therapeutic glycoproteins. AB - BACKGROUND: Patatins encoded by a multi-gene family are one of the major storage glycoproteins in potato tubers. Potato tubers have recently emerged as bioreactors for the production of human therapeutic glycoproteins (vaccines). Increasing the yield of recombinant proteins, targeting the produced proteins to specific cellular compartments, and diminishing expensive protein purification steps are important research goals in plant biotechnology. In the present study, potato patatins were eliminated almost completely via RNA interference (RNAi) technology to develop potato tubers as a more efficient protein expression system. The gene silencing effect of patatins in the transgenic potato plants was examined at individual isoform levels. RESULTS: Based upon the sequence similarity within the multi-gene family of patatins, a highly conserved target sequence (635 nts) of patatin gene pat3-k1 [GenBank accession no. DQ114421] in potato plants (Solanum tuberosum L.) was amplified for the construction of a patatin-specific hairpin RNAi (hpRNAi) vector. The CaMV 35S promoter-driven patatin hpRNAi vector was transformed into the potato cultivar Desiree by Agrobacterium-mediated transformation. Ten transgenic potato lines bearing patatin hpRNA were generated. The effects of RNA interference were characterized at both the protein and mRNA levels using 1D and 2D SDS/PAGE and quantitative real-time RT-PCR analysis. Dependent upon the patatin hpRNAi line, patatins decreased by approximately 99% at both the protein and mRNA levels. However, the phenotype (e.g. the number and size of potato tuber, average tuber weight, growth pattern, etc.) of hpRNAi lines was not distinguishable from wild-type potato plants under both in vitro and ex vitro growth conditions. During glycoprotein purification, patatin-knockdown potato tubers allowed rapid purification of other potato glycoproteins with less contamination of patatins. CONCLUSION: Patatin specific hpRNAi effectively suppressed the expression of a majority of patatin variants in potato tubers via the specific degradation of individual mRNAs of the patatin multi-gene family. More importantly, patatin-knockdown potato tubers appear to be an ideal host for the production of human therapeutic glycoproteins, because they eventually allow fast, easy purification of recombinant proteins, with less contamination from potato glycoprotein patatins. PMID- 18384694 TI - Design of price incentives for adjunct policy goals in formula funding for hospitals and health services. AB - BACKGROUND: Hospital policy involves multiple objectives: efficiency of service delivery, pursuit of high quality care, promoting access. Funding policy based on hospital casemix has traditionally been considered to be only about promoting efficiency. DISCUSSION: Formula-based funding policy can be (and has been) used to pursue a range of policy objectives, not only efficiency. These are termed 'adjunct' goals. Strategies to incorporate adjunct goals into funding design must, implicitly or explicitly, address key decision choices outlined in this paper. SUMMARY: Policy must be clear and explicit about the behaviour to be rewarded; incentives must be designed so that all facilities with an opportunity to improve have an opportunity to benefit; the reward structure is stable and meaningful; and the funder monitors performance and gaming. PMID- 18384695 TI - Measuring and explaining mortality in Dutch hospitals; the hospital standardized mortality rate between 2003 and 2005. AB - BACKGROUND: Indicators of hospital quality, such as hospital standardized mortality ratios (HSMR), have been used increasingly to assess and improve hospital quality. Our aim has been to describe and explain variation in new HSMRs for the Netherlands. METHODS: HSMRs were estimated using data from the complete population of discharged patients during 2003 to 2005. We used binary logistic regression to indirectly standardize for differences in case-mix. Out of a total of 101 hospitals 89 hospitals remained in our explanatory analysis. In this analysis we explored the association between HSMRs and determinants that can and cannot be influenced by hospitals. For this analysis we used a two-level hierarchical linear regression model to explain variation in yearly HSMRs. RESULTS: The average HSMR decreased yearly with more than eight percent. The highest HSMR was about twice as high as the lowest HSMR in all years. More than 2/3 of the variation stemmed from between-hospital variation. Year (-), local number of general practitioners (-) and hospital type were significantly associated with the HSMR in all tested models. CONCLUSION: HSMR scores vary substantially between hospitals, while rankings appear stable over time. We find no evidence that the HSMR cannot be used as an indicator to monitor and compare hospital quality. Because the standardization method is indirect, the comparisons are most relevant from a societal perspective but less so from an individual perspective. We find evidence of comparatively higher HSMRs in academic hospitals. This may result from (good quality) high-risk procedures, low quality of care or inadequate case-mix correction. PMID- 18384696 TI - Cultured stellate cells in human vocal fold mucosa. AB - OBJECTIVES: Stellate cells in the maculae flavae, located at both ends of the human vocal fold mucosa, have been considered an independent category of cells. We aimed to isolate and subculture these stellate cells, and to observe their morphological characteristics. METHODS: Stellate cells from the maculae flavae and fibroblasts from Reinke's space were cultured in three normal, adult human vocal fold mucosa preparations. RESULTS: The subcultured cells from Reinke's space were conventional fibroblasts. The subcultured cells from the maculae flavae were stellate in shape and had cytoplasmic processes. They were larger than conventional fibroblasts, and lipid droplets in the cytoplasm disappeared in the second culture. These stellate cells proliferated by attaching their cytoplasmic processes to each other. During the seven to 10 month subculture period, each cell type continued to exhibit its own morphological characteristics. CONCLUSION: This study demonstrated that such stellate cells form an independent cell category, which should be considered as a new category of cells within the human vocal fold. PMID- 18384697 TI - Retained knife blades in the ear, nose and throat: three cases. AB - OBJECTIVE: To discuss the management and to review the literature regarding retained knife blades in the head and neck. CASE REPORT: We present three cases in which patients presented with retained knife blades in the head and neck region; in two of these, the diagnosis was delayed by more than eight weeks. In all patients, the retained knife blade was removed through the pathway of insertion, without significant sequelae. DISCUSSION: The methods of removal, appropriate radiological investigations and patient profiles are discussed. CONCLUSIONS: We propose that radiography be performed on all patients presenting with facial stab injuries which are anything more than superficial. We further suggest that the direct extraction of sharp objects through the pathway of insertion is safe if radiological studies show no risk of vascular injury. PMID- 18384698 TI - Laryngeal reconstruction following shrapnel injury in a British soldier: case report. AB - INTRODUCTION: Laryngeal injuries are uncommon but result in high mortality and morbidity rates when they do occur. We report a case of laryngeal fracture due to penetrating shrapnel injury, repaired with miniplates. CASE REPORT: A 26-year-old soldier was involved in an explosion and sustained a shrapnel wound to his right neck. After immediate airway management at the field hospital he was transferred to the UK, where he underwent a neck exploration, laryngofissure and repair of the thyroid cartilage using miniplates. An endolaryngeal stent was placed, which was removed at a second operation seven days later. Post-operatively, the patient recovered well and his voice improved rapidly. Six months post-operatively, he returned to work. DISCUSSION: The cause and nature of laryngeal injury differs between wartime and peacetime. The methods of diagnosis and management strategies are reviewed. The early recognition of injury and protection of the airway are of paramount importance when dealing with laryngeal injury. Delayed laryngeal reconstruction using miniplates can give a good functional result. PMID- 18384699 TI - Front-line epistaxis management: let's not forget the basics. AB - OBJECTIVE: We postulated that epistaxis patients frequently have their nose packed in the accident and emergency department without any reasonable prior attempt at nasal assessment and nasal cautery. DESIGN: Telephone survey of 104 accident and emergency departments. RESULTS: A 100 per cent response rate was achieved. In all but one of the departments surveyed, first line management was carried out by accident and emergency doctors. Seventy-four per cent of accident and emergency doctors perform nasal cautery as first line management. Within accident and emergency departments, the availability and usage of head illumination, nasal speculums, topical anaesthesia and suction was grossly suboptimal. Only one-sixth of the accident and emergency doctors surveyed had received training in the management of epistaxis, whilst 81 per cent of accident and emergency departments would admit nasally packed patients under the ENT team. CONCLUSION: Despite the majority of accident and emergency doctors claiming to attempt nasal cautery before packing, this is unlikely to be effective if performed without the correct equipment. Better training and equipment need to be provided to accident and emergency doctors in order to optimise the management of epistaxis patients. This could potentially reduce inappropriate admissions. PMID- 18384700 TI - Sleep position and laterality of benign paroxysmal positional vertigo. AB - OBJECTIVE: The aim of this study was to investigate the frequency of posterior semicircular canal benign paroxysmal positional vertigo in each ear, and to assess the association between the ear affected by benign paroxysmal positional vertigo and the head-lying side during sleep onset. Based on a previous study which used objective methods to prove the preference of the elderly for the right head-lying side during sleep, we hypothesised that a predominance of the same head-lying side in benign paroxysmal positional vertigo patients may affect the pathophysiology of otoconia displacement. STUDY DESIGN: We conducted a prospective study of out-patients with posterior semicircular canal benign paroxysmal positional vertigo, confirmed by a positive Dix-Hallpike test. METHODS: One hundred and forty-two patients with posterior semicircular canal benign paroxysmal positional vertigo were interviewed about their past medical history, focusing on factors predisposing to benign paroxysmal positional vertigo. All patients included in the study were able to define a predominant, favourite head-lying side, right or left, during sleep onset. RESULTS: The Dix Hallpike test was found to be positive on the right side in 82 patients and positive on the left side in 54; six patients were found to be positive bilaterally. During sleep onset, 97 patients habitually laid their head on the right side and the remaining 45 laid their head on the left. The association between the affected ear and the head-lying side during sleep onset was statistically significant (p < 0.001). CONCLUSIONS: Our study found a predominance of right-sided benign paroxysmal positional vertigo, a subjective preference amongst patients for a right head-lying position during sleep onset, and an association between the ear affected by benign paroxysmal positional vertigo and the preferred head-lying side during sleep onset. The clinical and therapeutical implications of this observation are discussed. PMID- 18384701 TI - Complications after type one thyroplasty: is day-case surgery feasible? AB - OBJECTIVES AND HYPOTHESIS: Isshiki type one medialisation thyroplasty is an accepted treatment for a unilateral immobile vocal fold. It can also be performed simultaneously as a bilateral procedure in patients with severe bowing of the vocal folds (e.g. presbyphonia). The objectives of this study were to assess the incidence and timing of post-operative complications, and to evaluate whether patients undergoing this operation could, in future, be treated as day cases. STUDY DESIGN AND METHODS: A retrospective analysis was undertaken of 57 consecutive patients who had undergone a type one thyroplasty (52 unilateral and five bilateral) at a tertiary referral centre between April 2003 and April 2006. Post-operative improvement in the voice (measured subjectively, perceptually and quantitatively) was considered to constitute a successful outcome. Any complications were documented. RESULTS: Fifty-seven patients who had undergone laryngeal framework surgery were recruited from the study database. All of these patients had undergone either unilateral or bilateral type one medialisation thyroplasty but no arytenoid surgery. Thirty-seven were male (65 per cent) and 20 female (35 per cent), and there was left-sided predominance (74 per cent). All patients were discharged the morning following afternoon surgery (i.e. within 24 hours). Complications occurred in four patients (7 per cent). One patient, who was taking warfarin, developed a post-operative haematoma which resolved with conservative treatment. Two patients (both of whom had undergone revision thyroplasty) developed a wound infection three days post-operatively, which resolved with antibiotics. One patient returned with hoarseness five months post operatively, after an initially successful result. This patient had previously received radiotherapy for early glottic carcinoma, and the Silastic implant was eroding through the mucosa. This was subsequently removed under general anaesthesia. No patients developed complications leading to airway compromise. CONCLUSION: The only complications in this series were in patients taking anticoagulation medication, undergoing revision surgery, or in whom the laryngeal tissue was atrophic or absent. Careful patient selection to exclude any of the above should reduce the risk of complications. The authors would therefore advocate type one thyroplasty for unilateral or bilateral vocal fold paralysis as a suitable procedure for day-case surgery within our department. PMID- 18384702 TI - A potential danger of flexible endoscopy sheaths: a detached tip and how to retrieve it. AB - OBJECTIVE: To report an unrecognised complication of fibre-optic nasendoscopy, and its management. CASE REPORT: A protective, transparent nasendoscopy sheath is often used to reduce nasendoscope 'downtime' and to prevent cross infection, with minimal effect on the obtained image quality. We report the case of a subcutaneous tracheostomy procedure during which, without undue strain, the tip of the sheath became detached and acted as a foreign body within the trachea. A urological stone retrieval basket was used to retrieve the sheath, after failure of conventional methods. DISCUSSION: Clinicians should be aware that any instrument introduced into the airway has the potential to fail and in the process produce a foreign body which may cause serious complications. The urological stone retrieval basket may be a useful addition to the current set of instruments used to deal with difficult airway foreign bodies. PMID- 18384703 TI - Synergic effect of vitamin A and high-fat diet in adipose tissue development and nuclear receptor expression in young rats. AB - In order to study the effects of dietary lipids and vitamin A on the development of adipose tissues, young rats were submitted for 8 d to a control or to two cafeteria diets with normal (Caf) or higher (Caf + ) vitamin A levels. Retinoid (retinoic acid receptor (RAR) a, RARg, retinoid X receptor(RXR) alpha) and fatty acid (PPARgamma) receptor mRNA was measured in the subcutaneous white adipose tissue (Swat) and in isolated mature adipocytes by RT-PCR. The stroma vascular fraction was cultured in vitro to test the capacities of the adipocyte precursors to proliferate and differentiate.The Caf diet enriched in vitamin A resulted in an increased adiposity, due to increased adipocyte hypertrophy. This was concomitant with a lower expression of RARa and RARg mRNA (234.6 and 238.6 %) and a higher expression of PPARgamma (+59 %) in the Swat and, to a less extent,in isolated adipocytes. Positive correlations were obtained between PPARgamma mRNA and Swat weights and between PPARgamma and RXRalpha mRNA. By contrast, RARgamma mRNA and Swat masses were negatively correlated. The adipocyte precursors from Caf + Swat proliferated more,in vitro, at the beginning of the culture. This difference progressively disappeared and was totally absent after 8 d of culture, but with a higher percentage of differentiated preadipocytes (+80.3 %) in the Caf + group. In conclusion, lipids and vitamin A act synergistically on the normal growth of the adipose tissue in young rats, concomitant with an imbalance in the pattern of the nuclear receptors. These changes influence the early normal development of the endogenous adipocyte precursors. PMID- 18384704 TI - Expanded adipose tissue: 'out of breath' and inflamed. PMID- 18384706 TI - Non-response to consecutive antidepressant therapy caused by CYP2D6 ultrarapid metabolizer phenotype. PMID- 18384705 TI - Consumption of fructose-sweetened beverages for 10 weeks increases postprandial triacylglycerol and apolipoprotein-B concentrations in overweight and obese women. AB - Fructose consumption in the USA has increased over the past three decades. During this time, obesity, insulin resistance and the metabolic syndrome have also increased in prevalence. While diets high in fructose have been shown to promote insulin resistance and increase TAG concentrations in animals, there are insufficient data available regarding the long-term metabolic effects of fructose consumption in humans. The objective of the present study was to investigate the metabolic effects of 10-week consumption of fructose-sweetened beverages in human subjects under energy-balanced conditions in a controlled research setting. Following a 4-week weight-maintaining complex carbohydrate diet, seven overweight or obese (BMI 26.8-33.3 kg/m2) postmenopausal women were fed an isoenergetic intervention diet, which included a fructose-sweetened beverage with each meal, for 10 weeks. The intervention diet provided 15 % of energy from protein, 30 % from fat and 55 % from carbohydrate (30 % complex carbohydrate, 25 % fructose). Fasting and postprandial glucose, insulin, TAG and apoB concentrations were measured. Fructose consumption increased fasting glucose concentrations and decreased meal-associated glucose and insulin responses (P = 0.0002, P = 0.007 and P = 0.013, respectively). Moreover, after 10 weeks of fructose consumption, 14 h postprandial TAG profiles were significantly increased, with the area under the curve at 10 weeks being 141 % higher than at baseline (P = 0.04). Fructose also increased fasting apoB concentrations by 19 % (P = 0.043 v. baseline). In summary, consumption of fructose-sweetened beverages increased postprandial TAG and fasting apoB concentrations, and the present results suggest that long-term consumption of diets high in fructose could lead to an increased risk of CVD. PMID- 18384707 TI - Definition of difficult tracheal intubation. PMID- 18384708 TI - The role of healthcare communications agencies in maintaining compliance when working with the pharmaceutical industry and healthcare professionals. AB - BACKGROUND: Relationships between the pharmaceutical industry and healthcare professionals continue to drive discussion about the potential for conflicts of interest. Despite greater regulation and oversight, there are still calls for increased transparency and further restrictions on these relationships. Regulatory authorities, the pharmaceutical industry, professional societies, and other interested parties have responded by developing robust guidelines for interactions between the pharmaceutical industry and healthcare professionals. This, in turn, is driving change in the way that healthcare communications agencies work, increasing the need for them to visibly demonstrate processes that ensure their employees comply with relevant laws, regulations, and guidelines. EXPOSITION: In our group of healthcare communications agencies we have established an internal compliance program and developed a policy that reflects the services we provide, and we recommend that other agencies adopt a similar program. Compliance training, implemented by a nominated compliance team, can be enforced by including compulsory tests for employees who interact with the pharmaceutical industry and healthcare professionals, with annual reassessment. The compliance team also has an important role to play in ensuring ongoing communication and staff education, including awareness of new legal and best practice developments. Management of the compliance program is essential, with clear mechanisms for auditing and evaluation, and the inclusion of compliance adherence in staff performance objectives. A visible framework for handling potential compliance issues should also be developed, with clear definitions of different levels of noncompliance and potential associated consequences. Compliance programs may also include other elements, such as terminology and documentation guidance, so that the program becomes an integral tool used by employees on a daily basis. CONCLUSION: With a robust internal compliance program, healthcare communication agencies can play a significant role in helping maintain appropriate pharmaceutical industry-healthcare professional relationships in an increasingly regulated and scrutinized environment. PMID- 18384709 TI - A qualitative review of issues arising in the use of psycho-stimulant medications in patients with ADHD and co-morbid substance use disorders. AB - OBJECTIVE: This review addresses the relationship between attention deficit/hyperactivity disorder (ADHD) and substance use disorders (SUDs), with an emphasis on factors that determine the potential for psychostimulant abuse. Strategies for identification and treatment of patients with ADHD who are at risk for, or have, co-morbid SUD are also addressed. RESEARCH DESIGN AND METHODS: The article was based on a qualitative review of current literature addressing co morbid ADHD and SUD. DISCUSSION: Adolescent and adult patients with ADHD are at increased risk for SUD, as well as a number of other psychiatric disorders. Psychostimulant agents like methylphenidate (MPH) and mixed amphetamine salts (MAS) are effective first-line pharmacotherapies for ADHD; however, they are Schedule II controlled substances with a potential for abuse. Evidence suggests that treatment of ADHD during childhood with stimulant agents may reduce the risk of developing SUD later on. Factors associated with the highest risk of SUD in patients with ADHD include co-morbid antisocial personality disorder, bipolar disorder, an eating disorder, severe ADHD and/or antisocial behavior symptoms, and dropping out of school. Treatment initiation during adolescence or young adulthood also has been linked to increased risk of polydrug use and non-medical stimulant use, a pattern of behavior consistent with a risk of SUD development. Treatment plans for patients with ADHD and co-morbid SUD should include behavioral interventions, careful monitoring, and when appropriate, pharmacotherapy. When oral formulations of psychostimulants are used at recommended doses and frequencies, they are unlikely to yield effects consistent with abuse potential in patients with ADHD. Long-acting stimulant formulations and non-stimulants, like atomoxetine or bupropion, have a lower potential for abuse, and provide several safe and effective treatment options for the development of a comprehensive management plan for patients with co-morbid ADHD and SUD. CONCLUSIONS: The present review is neither exhaustive nor systematic. Moreover, the reviewed studies vary widely with regards to methodology and patient populations. In light of these limitations, several conclusions are still warranted. Patients with ADHD are at increased risk for SUD. Under certain conditions, psychostimulants may be a pharmacologic option in the treatment of patients with co-morbid ADHD and SUD. However, clinicians should be mindful of the risks and benefits of this treatment approach in a high-risk population and should also bear in mind the labeling guidelines when working with this co morbidity. PMID- 18384710 TI - The effect of statins on the development of new-onset type 2 diabetes: a meta analysis of randomized controlled trials. AB - OBJECTIVE: To determine the ability of statins to prevent the development of new onset type 2 diabetes mellitus through a meta-analysis of randomized, controlled trials. RESEARCH DESIGN AND METHODS: A systematic literature search through November 6, 2007 was conducted to identify randomized, placebo-controlled trials of statins that reported data on the incidence of new-onset diabetes mellitus. Incidence of new-onset type 2 diabetes mellitus was treated as a dichotomous variable. Weighted averages were reported as relative risk (RR) with associated 95% confidence intervals (CI). A random-effects model was used. RESULTS: Five prospective, randomized controlled trials (n = 39,791) were identified. Upon meta analysis, the use of a statin did not significantly alter a patient's risk of developing new-onset type 2 diabetes mellitus (relative risk, 1.03; 95% confidence interval 0.89-1.19). Subgroup and sensitivity analyses did not significantly change the results. There was statistical heterogeneity that stemmed from pravastatin's tendency towards a reduction in risk and the other statins showing an increase in risk. The funnel plot could not rule out publication bias. CONCLUSIONS: Statins, as a class, do not demonstrate a statistically significant positive or negative impact on a patient's risk of developing new-onset type 2 diabetes mellitus. PMID- 18384711 TI - Efficacy of risedronate against hip fracture in patients with neurological diseases: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: Neurological diseases, including Alzheimer's disease, stroke, and Parkinson's disease have been reported to increase the risk for fractures. The purpose of the present study was to clarify the efficacy of risedronate against hip fracture in patients with neurological diseases. METHODS: The literature was searched with PubMed from 1995 to the present, with respect to strictly conducted randomized controlled trials (RCTs) with narrow confidence intervals (CIs), and a meta-analysis was conducted. RESULTS: Four RCTs met the criteria; one RCT for Alzheimer's disease (n = 461, mean age = 78 years), two RCTs for stroke (n = 267, mean age = 76 years for men; n = 345, mean age = 71 years for women), and one RCT for Parkinson's disease (n = 223, mean age = 71 years). According to the results of RCTs, the relative risks (95% CI) for hip fracture with risedronate treatment compared with placebo treatment were 0.26 (0.10, 0.69) for Alzheimer's disease, 0.20 (0.04, 0.89) for stroke in men, 0.14 (0.02, 1.16) for stroke in women, and 0.34 (0.09, 1.21) for Parkinson's disease. Overall, the relative risk (95% CI) for hip fracture with risedronate treatment was 0.25 (0.13, 0.48), suggesting 75% of risk reduction rate with risedronate treatment in patients with one of the three neurological diseases (heterogeneity: 0.58, p = 0.9016 and overall effect: 17.36, p < 0.0001). No severe adverse events were reported in the risedronate and placebo groups. CONCLUSION: The results of a meta-analysis of strictly conducted RCTs suggest that there is efficacy against hip fracture and safety with risedronate treatment in patients with neurological diseases including Alzheimer's disease, stroke, and Parkinson's disease. PMID- 18384712 TI - Silent cerebral microbleeds on susceptibility-weighted imaging of patients with ischemic stroke and leukoaraiosis. AB - OBJECTIVE: Cerebral microbleeds (CMBs) are an increasingly recognized in patients with cerebrovascular disease who undergo the new MRI techniques. Susceptibility weighted MRI is sensitive to detect silent microbleeds which are associated with microangiopathy from atherosclerosis, amyloid angiopathy or asymptomatic elderly individuals. CMBs are intracerebral old hemosiderin deposits in the perivascular space, and the clinical significance of such 'microbleeds' remains controversy. We investigated the incidence of microbleeds among different ischemic stroke subtypes and the severity of leukoaraiosis. METHODS: In this study, we collected 137 patients (73.5 +/- 9.1 years old, 84 men and 53 women) who were divided into five groups: atherothrombotic infarction (34 patients), cardioembolic infarction (24 patients), lacunar infarction (35 patients), other determined or undetermined infarction (21 patients) and control (23 patients), in which 52 had primary stroke and 62 had recurrent stroke. Microbleeds were counted and leukoaraiosis was graded using susceptibility-weighted, T1-, T2- and FLAIR MRI with a 3.0 T system. RESULTS: Microbleeds were high in lacunar infarction (25.7%) and atherothrombotic infarction patients (20.6%). Microbleeds were low in patients with cardioembolic infarction (4.2%). Both microbleeds (> or =grade 2) and leukoaraiosis in severity (> or =grade 2) were higher in the recurrent stroke group (14.5 and 48.4%) than those in the primary stroke group (3.8 and 7.7%). Leukoaraiosis (r=0.803, p<0.05) and recurrent stroke (r=0.708, p<0.05) were significantly associated with microbleeds. CONCLUSIONS: Asymptomatic CMBs are observed with a markedly higher frequency in recurrent stroke, and are closely associated with the severity of leukoaraiosis. PMID- 18384713 TI - New method to study DNA sequences: the languages of evolution. AB - Recently, several authors have reported statistical evidence for deterministic dynamics in the flux of genetic information, suggesting that evolution involves the emergence and maintenance of a fractal landscape in DNA chains. Here we examine the idea that motif repetition lies at the origin of these statistical properties of DNA. To analyse repetition patterns we apply a modification of the BDS statistic, devised to analyze complex economic dynamics and adapted here to DNA sequence analysis. This provides a new method to detect structured signals in genetic information. We compare naturally occurring DNA sequences along the evolutionary tree with randomly generated sequences and also with simulated sequences with repetition motifs. For easier understanding, we also define a new statistic for a DNA sequence that constitutes a specific fingerprint. The new methods are applied to exon and intron DNA sequences, finding specific statistical differences. Moreover, by analysing DNA sequences of different species from Bacteria to Man, we explore the evolution of these linguistic DNA features along the evolutionary tree. The results are consistent with the idea that all the flux of DNA information need not be random, but may be structured along the evolutionary tree. The implications for evolutionary theory are discussed. PMID- 18384714 TI - Dimensional analysis of actigraphic derived sleep data. AB - Nonlinear dimensional analyses can be a useful tool in understanding the underlying behavior of dynamical systems, including biological systems. Many biological functions can be modeled as chaotic processes, including sleep. Sleep data can be obtained from several methods, such as electroencephalograms, polysomnography, and actigraph. Actigraphy, because of its low level of invasiveness, is an increasingly popular method of obtaining sleep data. This study analyzed actigraphy data with nonlinear dimensional analyses to determine if such analytic methods would be useful in sleep studies. Participants wore actigraphs on their wrists, which recorded movement for several days. Several sleep quality variables, such as movement during sleep and total sleep time, were derived from these sleep data. These variables were used to determine whether the quality of sleep was good or poor. Lagged phase space plots were graphed and nonlinear parameters for the fractal dimension and the correlation dimension were computed for each participant. Descriptive and inferential statistics were performed to determine if the nonlinear parameters showed significant differences with respect to sleep quality. PMID- 18384715 TI - Entropy conservation in the control of human action. AB - The human motor system is highly adaptable with the ability to adjust its movement patterns under constantly changing task and environmental constraints. In this paper we develop the position that the probabilistic nature of human action can be characterized by entropies at the level of the organism, task, and environment. Systematic changes in motor adaptation are characterized as task organism and environment-organism tradeoffs in entropy. Such compensatory adaptations lead to a view of goal-directed motor control as the product of an underlying conservation of entropy across the task-organism-environment system. The conservation of entropy supports the view that context dependent adaptations in human goal-directed action are guided fundamentally by natural law and provides a novel means of examining human motor behavior. PMID- 18384716 TI - Dynamic constancy as a basis for perceptual hierarchies. AB - Bateson's difference-based epistemology can be simulated by a Boolean network model. Bateson proposed that taking differences in differences would produce emergent hierarchies of knowledge. This study simulated Bateson's proposal by taking differences in differences in a Boolean model. The crucial result is that constancies in the dynamics of the flow of differences in the model (a) define perceptually comprehensible categories of visual forms and (b) that higher-order constancies arrange these categories into a perceptually comprehensible hierarchy. We propose Dynamic Constancy as a new Gestalt-like organizational principle in perception. PMID- 18384717 TI - Cusp catastrophe model for binge drinking in a college population. AB - A cusp catastrophe model for binge drinking behavior was developed and tested with attitude toward alcohol consumption and peer influence as the two control parameters. Similar models were also developed for frequency and quantity of alcohol use. Participants were 1,247 students who completed the Long Form of the Core Alcohol and Drug Survey. The results were strongest for the binge drinking criterion (R(2) = .90), compared to a linear model (R(2) = .34) that is usually associated with the Theory of Planned Behavior or Theory of Reasoned Action. The results have numerous implications for the development of interventions and for future research. PMID- 18384718 TI - Radiolabeling rhesus monkey CD34+ hematopoietic and mesenchymal stem cells with 64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) for microPET imaging. AB - Noninvasive positron emission tomography (PET) provides a potential method for in vivo tracking of radiolabeled cells. The goal of this study was to assess the potential toxicity of 64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) (PTSM) on rhesus monkey CD34+ hematopoietic and mesenchymal stem cells in vitro in preparation for developing imaging protocols posttransplantation. CD34+ hematopoietic cells were radiolabeled with 0 to 40 microCi/mL 64Cu-PTSM and viability and colony formation were assessed. Rhesus monkey mesenchymal stem cells (rhMSCs) were placed in culture postradiolabeling for assessments of growth and differentiation toward adipogenic, osteogenic, and chondrogenic lineages. The results indicated that CD34+ cells radiolabeled with 20 microCi/mL and rhMSCs radiolabeled with 10 microCi/mL 64Cu-PTSM did not result in adverse effects on growth or differentiation. Nonradioactive copper was also evaluated and showed that the presence of copper was not harmful to the cells. CD34+ cells and rhMSCs radiolabeled with the optimized concentrations of 20 and 10 microCi/mL, respectively, were also assessed using the microPET scanner. Studies showed that a minimum of 2.50x10(4) CD34+ cells (1.1 pCi/cell) and 6.25x10(3) rhMSCs (4.4 pCi/cell) could be detected. These studies indicate that CD34+ hematopoietic cells and rhMSCs can be safely radiolabeled with 64Cu-PTSM without adverse cellular effects. PMID- 18384719 TI - Use of step-section histopathology to evaluate 18F-fluorocholine PET sextant localization of prostate cancer. AB - To assess positron emission tomography (PET) with fluorine-18 fluorocholine for sextant localization of malignant prostate tumors. Histopathologic analysis was performed on step-sectioned whole-mounted prostate specimens from 15 patients who underwent PET with fluorocholine prior to radical prostatectomy. The maximum standardized uptake value (SUVmax) corresponding to prostate sextants on PET was measured by region of interest analysis and compared with histopathologic results. Histopathology demonstrated malignant involvement in 61 of 90 prostate sextants. The mean total tumor volume per specimen was 4.9 mL (range 0.01-28.7 mL). Mean SUVmax was 6.0+/-2.0 in malignant sextants and 3.8+/-1.4 in benign sextants (p<.0001). The area under the receiver operating characteristic curve was 0.82 for sextant detection of malignancy based on SUVmax measurement. Tumor diameter directly correlated with sextant SUVmax in malignant sextants (r=.54, p<.05). In 13 subjects, the largest tumor in the specimen corresponded to the sextant with the highest SUVmax. Fluorocholine PET can serve to localize dominant areas of malignancy in patients with prostate cancer. However, PET with fluorocholine may fail to identify sextants with smaller volumes of malignancy. PMID- 18384720 TI - Diet and abdominal autofluorescence detected by in vivo fluorescence imaging of living mice. AB - We investigated the effect of diet on abdominal autofluorescence detected by in vivo fluorescence imaging (FLI) of living mice. Groups of mice were fed a regular, alfalfa-free, or purified diet, and whole-body FLI was performed without the administration of fluorescent probes. In addition, quantum dots were injected intravenously into mice fed one of the three diets, and FLI was performed 3 and 24 hours later. Intense autofluorescence originating from the animals' intestinal contents was observed in mice fed the regular diet. Intestinal autofluorescence decreased substantially after feeding with the alfalfa-free diet and further after feeding with the purified diet. The decline was rapid and took only 1 to 2 days; however, it may have been affected by an intake of feces. The reticuloendothelial system was clearly delineated using a low dose of quantum dots in mice fed the purified diet. On the other hand, intestinal autofluorescence was visible 24 hours postinjection in mice given the alfalfa free diet and definitely impaired the image quality in mice fed the regular diet. The use of a low-fluorescence diet, especially a purified diet, rapidly reduces intestinal autofluorescence and is expected to enhance the potential of in vivo FLI. PMID- 18384721 TI - In vivo optical imaging of neurogenesis: watching new neurons in the intact brain. AB - Adult neurogenesis is a highly dynamic process modulated by several pathologic and environmental factors, as well as by various compounds. So far, available techniques to study neurogenesis are lengthy and personnel and cost intensive. We developed a new tool based on the doublecortin promoter driving the expression of the luciferase reporter gene (DCX-promo-luciferase) in transgenic mice to perform in vivo imaging of neurogenesis. Indeed, the DCX-promo-luciferase mice allowed optical in vivo imaging of the onset of and increase in neurogenesis in developing fetal brains, as well as imaging of neurogenesis in the intact adult mouse central nervous system. Moreover, the capacity to specifically detect a small number of migrating neuronal precursors in vivo after transplantation is for the first time feasible using this DCX-promo-luciferase transgenic tool. The present imaging approach offers several crucial advantages over methods currently available, such as bromodeoxyuridine incorporation or labeling using iron oxide nanoparticles. Hence, it allows longitudinal study of neurogenesis in intact animals without the requirement of cellular prelabeling. Moreover, it guarantees that detection is specific for neuronal precursors and restricted to viable cells. Hence, our DCX-promo-luciferase transgenic model constitutes an effective tool that answers the pressing need for rapid investigation of the impact on neurogenesis of a large number of candidate compounds waiting to be tested. PMID- 18384722 TI - Noninvasive in vivo imaging of protein kinase A activity. AB - Protein kinases play pivotal roles in almost all cellular signaling pathways, and modulation of their activity is desirable in both disease and studies of function. Information on the activity of kinases in vivo is scarce owing to a lack of appropriate methods. To obtain such information, we produced mice in which protein kinase A (PKA) activity can be monitored noninvasively in vivo. The model uses luciferase, which has been mutated to contain a target sequence of PKA, thus making luminescence from the enzyme dependent on its state of phosphorylation. The PKA-sensitive luciferase, termed luciferase(PKA), was incorporated into the mouse genome, and transgenic animals exhibited a rapid beta adrenergic response, that is, reduced luminescence, in various organs, including the pancreas, muscle, liver, and fat, after isoproterenol injection. This study shows that luciferase can be used for in vivo measurements of kinase activity, suggesting that different kinase target sequences in luciferase can monitor kinase activity modulation. PMID- 18384723 TI - Visualizing head and neck tumors in vivo using near-infrared fluorescent transferrin conjugate. AB - Transferrin receptor (TfR) is overexpressed in human head and neck squamous cell carcinomas (HNSCCs). This study was carried out to investigate the feasibility of imaging HNSCC by targeting TfR using near-infrared fluorescent transferrin conjugate (TfNIR). Western blot analysis of four HNSCC cell lines revealed overexpression of TfR in all four lines compared with that in normal keratinocytes (OKFL). Immunocytochemistry further confirmed the expression of TfR and endocytosis of TfNIR in JHU-013 culture cells. Following intravenous administration of TfNIR (200 microL, 0.625 microg/microL), fluorescent signal was preferentially accumulated in JHU-013 tumor xenografts grown in the lower back (n=14) and oral base tissues (n=4) of nude mice. The signal in tumors was clearly detectable as early as 10 minutes and reached the maximum at 90 to 120 minutes postinjection. The background showed an increase, followed by a decrease at a much faster pace than tumor signal. A high fluorescent ratio of the tumor to muscle was obtained (from 1.42 to 4.15 among tumors), usually achieved within 6 hours, and correlated with the tumor size (r=.74, p=.002). Our results indicate that TfR is a promising target and that Tf(NIR)-based optical imaging is potentially useful for noninvasive detection of early HNSCC in the clinic. PMID- 18384725 TI - Identification of potentially hazardous human gene products in GMO risk assessment. AB - Genetically modified organisms (GMOs), e.g. viral vectors, could threaten the environment if by their release they spread hazardous gene products. Even in contained use, to prevent adverse consequences, viral vectors carrying genes from mammals or humans should be especially scrutinized as to whether gene products that they synthesize could be hazardous in their new context. Examples of such potentially hazardous gene products (PHGPs) are: protein toxins, products of dominant alleles that have a role in hereditary diseases, gene products and sequences involved in genome rearrangements, gene products involved in immunomodulation or with an endocrine function, gene products involved in apoptosis, activated proto-oncogenes. For contained use of a GMO that carries a construct encoding a PHGP, the precautionary principle dictates that safety measures should be applied on a "worst case" basis, until the risks of the specific case have been assessed. The potential hazard of cloned genes can be estimated before empirical data on the actual GMO become available. Preliminary data may be used to focus hazard identification and risk assessment. Both predictive and empirical data may also help to identify what further information is needed to assess the risk of the GMO. A two-step approach, whereby a PHGP is evaluated for its conceptual dangers, then checked by data bank searches, is delineated here. PMID- 18384724 TI - Gold nanorods targeted to delta opioid receptor: plasmon-resonant contrast and photothermal agents. AB - Molecularly targeted gold nanorods were investigated for applications in both diagnostic imaging and disease treatment with cellular resolution. The nanorods were tested in two genetically engineered cell lines derived from the human colon carcinoma HCT-116, a model for studying ligand-receptor interactions. One of these lines was modified to express delta opioid receptor (deltaOR) and green fluorescent protein, whereas the other was receptor free and expressed a red fluorescent protein, to serve as the control. Deltorphin, a high-affinity ligand for deltaOR, was stably attached to the gold nanorods through a thiol-terminated linker. In a mixed population of cells, we demonstrated selective imaging and destruction of receptor-expressing cells while sparing those cells that did not express the receptor. The molecularly targeted nanorods can be used as an in vitro ligand-binding and cytotoxic treatment assay platform and could potentially be applied in vivo for diagnostic and therapeutic purposes with endoscopic technology. PMID- 18384726 TI - Effects of rhizodeposition of non-transgenic and transplastomic tobaccos on the soil bacterial community. AB - The effect of root-released compounds of transplastomic tobacco (Nicotiana tabacum) on the soil bacterial community structure, and their potential to support horizontal gene transfer (HGT) to bacteria have been studied. Soil microcosms were exposed to root-released compounds collected from transplastomic and non-transgenic tobacco cultivars. Cluster analysis of automated ribosomal intergenic spacer analysis (ARISA) profiles of the soil bacterial community after 48 h incubation grouped the transgenic cultivar apart from the non-transgenic, indicating that it had a rhizodeposition pattern different from the parental plants. However, these differences were less than between the two non-transgenic tobacco cultivars studied. NMR characterization of the root-released compounds showed some differences in chemical fingerprinting pattern between the transplastomic and the parental cultivar. However, the effect on bacterial community structure was transient, and tended to disappear after 96 h of incubation. The potential of root-released compounds as a source of transforming DNA for bacteria was investigated by using four potential recipient species. No transformants were obtained following exposure of all the recipients to the root released compounds. Root-released compounds amended to transgene donor DNA decreased the transformation frequency of Acinetobacter baylyi strain ADP1200, while Azospirillum, Agrobacterium, and Sinorhizobium strains failed to develop competence also in the presence of an external added transgene source. Detection of plastid sequences by PCR suggested that a very low amount of fragmented plastid donor DNA was present in the root-released compounds. PMID- 18384727 TI - Exploration of methods used to describe bacterial communities in silage of maize (Zea mays) cultivars. AB - Different techniques to assess bacterial community structure and diversity were evaluated in silages prepared with four different maize cultivars, three conventional and one transgenic (cv. Tundra, event Bt-176). Plants were cultivated in the greenhouse and harvested after 30 days of growth. Silage samples were collected at successive times during fermentation and analyzed for bacterial counts and by various DNA-based fingerprinting techniques. Bacterial counts were similar between cultivars for the total culturable bacteria, sporeforming, and mesophilic and thermophilic lactic acid bacteria (LAB). Further analysis of the species composition of 388 LAB strains by intergenic transcribed spacer (ITS) PCR followed by sequencing of 16S rRNA gene did not reveal differences between cultivars. In contrast, molecular fingerprinting methods targeting whole bacterial communities, such as automated ribosomal intergenic spacers analysis (ARISA) and 16S rRNA gene length heterogeneity-PCR (LH-PCR), indicated that different maize silage batches or cultivars hosted different bacterial communities. Thus, ARISA and LH-PCR fingerprinting techniques offer a fast and sensitive method to compare bacterial communities, and to detect differences in silage bacterial communities. PMID- 18384728 TI - A screening method for prioritizing non-target invertebrates for improved biosafety testing of transgenic crops. AB - We have developed a screening method that can be used during the problem formulation phase of risk assessment to identify and prioritize non-target invertebrates for risk analysis with any transgenic plant. In previously published protocols for this task, five criteria predominated. These criteria have been combined by our method in a simple model which assesses: (1) the possible level of risk presented by the plant to each invertebrate species (through measurements of potential hazard and exposure, the two principal criteria); (2) the hypothetical environmental impact of this risk (determined by the currently known status of the species' population in the ecosystem and its potential resilience to environmental perturbations); (3) the estimated economic, social and cultural value of each species; and (4) the assessed ability to conduct tests with the species. The screening method uses information on each of these criteria entered into a specially designed database that was developed using Microsoft Access 2003. The database holds biological and ecological information for each non-target species, as well as information about the transgenic plant that is the subject of the risk assessment procedure. Each piece of information is then ranked on the basis of the value of the information to each criterion being measured. This ranking system is flexible, allowing the method to be easily adapted for use in any agro-ecosystem and with any plant modification. A model is then used to produce a Priority Ranking of Non-Target Invertebrates (PRONTI) score for each species, which in turn allows the species to be prioritized for risk assessment. As an example, the method was used to prioritize non-target invertebrates for risk assessment of a hypothetical introduction of Bacillus thuringiensis (Bt) Cry1Ac-expressing Pinus radiata trees into New Zealand. PMID- 18384729 TI - Perspectives on the ecological assessment of transitional waters. PMID- 18384734 TI - Sodium-deoxycholate-assisted tryptic digestion and identification of proteolytically resistant proteins. AB - Identification of proteolytically resistant proteins with compact molecular structure and/or poor water solubility is a challenge in current proteomic study. In this study, sodium deoxycholate (SDC)-assisted tryptic digestion and identification of proteolytically resistant myoglobin and integral membrane proteins were systematically investigated. When the effect of SDC up to 10% on trypsin activity was investigated, little decrease in the trypsin activity was observed in 1% SDC solution, 2-5% SDC decreased the enzyme activity only by about 13.6%, and even in the presence of 10% SDC trypsin still retained 77.4% of its activity. Matrix-assisted laser desorption ionization time of flight mass spectrometry analysis showed that SDC could be removed from sample solution with acid treatment followed by centrifugation, and the remaining SDC, if any, had little effect on mass spectrometry analysis with regard to the number and signal/noise ratio of ions in the mass spectra. Compared with urea and methanol, two other commonly used additives in addition to SDS in proteomic analysis, SDC improved more efficiently the denaturation, solubilization, and tryptic digestion of proteins, particularly proteolytically resistant myoglobin and integral membrane proteins, thereby enhancing the efficiency of their identification with regard to the number of identified proteins and unique peptides and the sequence coverage of matched proteins. PMID- 18384735 TI - Strategy combining separation of isotope-labeled unfolded proteins and matrix assisted laser desorption/ionization mass spectrometry analysis enables quantification of a wide range of serum proteins. AB - A novel strategy for the quantitative profiling of serum proteome is described. It includes an ammonium sulfate depletion of the serum, an affordable stable isotope labeling chemistry for samples with a large amount of protein, separation of the unfolded proteins, and relative quantification by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). Labeling of unfolded proteins was performed using normal (D(0)) acrylamide and deuterated (D(3)) acrylamide. The workflow for separating the unfolded proteins includes whole gel elution and ion exchange liquid chromatography, and it combines electrophoretic separation based on the protein molecular weight followed by chromatographic separation in the presence of 8M urea based on protein charge. This was followed by trypsinolysis and MALDI MS analysis, leading to the quantification of a large number of serum proteins, including those with an abundance of 10(-5) less than albumin. This robust and inexpensive workflow is suitable for the quantitative profiling of protein changes in serum associated with preanalytical variables. PMID- 18384736 TI - A methylation-specific and SYBR-green-based quantitative polymerase chain reaction technique for O6-methylguanine DNA methyltransferase promoter methylation analysis. AB - The O(6)-methylguanine DNA methyltransferase (MGMT) gene encodes a DNA repair enzyme whose activity is a major mechanism of resistance to alkylating drugs in glioblastoma treatment. Hypermethylation of the MGMT promoter is associated with chemosensitivity because it reduces MGMT activity. Here we present a method combining methylation-specific and SYBR-green-based quantitative PCR (MSQP) for MGMT promoter methylation analysis. This highly specific, sensitive, and reproducible method allows the quantification of fully methylated and fully unmethylated MGMT DNA species in terms of percentage. Values are related to standard curves, corrected for DNA input by an internal standard, and calculated in relation to methylated and unmethylated control DNAs as a percentage share. Finally, values are defined relative to the sum of fully methylated and unmethylated MGMT DNA sample amount to obtain percentage of methylated reference and percentage of unmethylated reference results. We have used this technique to investigate MGMT promoter methylation in relation to MGMT mRNA expression in nine tumor cell lines and 15 primary glioblastoma patients. Presented data confirm that this assay is suitable for detection of low amounts of methylated and unmethylated MGMT promoter DNA. Carefully validated quantitative MSQP assays will be useful in both research and clinical molecular diagnosis. PMID- 18384737 TI - Influence of intermediate aminodextran layers on the signal response of surface acoustic wave biosensors. AB - Surface acoustic wave (SAW) devices based on horizontally polarized surface shear waves enable direct and label-free detection of proteins in real time. Binding reactions on the sensor surface are detected by determining changes in surface wave velocity caused mainly by mass adsorption or change of viscoelasticity in the sensing layer. Intermediate hydrogel layers have been proven to be useful to immobilize capture molecules or ligands corresponding to the analyte. However, the SAW signal response strongly depends on the morphology of the hydrogel due to different relative changes of its acoustomechanical parameters such as viscoelasticity and density. In this work five aminodextrans (AMD) and one diamino polyethylene glycol (DA-PEG) were used as intermediate hydrogel layers. Sensors with immobilized streptavidin and samples containing biotinylated bovine serum albumin were used to exemplify affinity assays based on immobilized capture molecules for protein detection. The effects of the three-dimensional AMDs and the two-dimensional (2D) DA-PEG on the SAW signal response were investigated. The signal height decreased with increasing molar mass and increasing amount of immobilized AMD. Consequently, thin hydrogel layers are ideal to obtain optimum signal responses in this type of assay, whereas it is not necessarily a 2D hydrogel that gives the best results. PMID- 18384738 TI - Investigating human immunodeficiency virus-1 proteinase specificity at positions P4 to P2 using a bacterial screening system. AB - Inhibitors of human immunodeficiency virus-1(HIV-1) proteinase have been used for several years to treat acquired immunodeficiency syndrome patients. Despite intensive research, however, the substrate specificity of this enzyme is not completely elucidated. Here, we assessed the HIV-1 proteinase P(4) to P(2) substrate specificity using a bacterial screening system. In this system, the bacterial enzyme beta-galactosidase has been transformed into an HIV-1 proteinase substrate by insertion of the p6/PR cleavage site. Consequently, HIV-1 processing can be determined by measuring the beta-galactosidase activity on X-gal plates and by examination of the extent of cleavage of the beta-galactosidase protein itself. We screened a library containing randomized sequences at the P(4) to P(2) positions and found strong preferences for Thr, Ser, and Pro at P(4), for Leu, Met, and Phe at P(3), and for Ser, Met, and Leu at P(2). The frequent observations of Thr at P(4) and Ser at P(2) extend previous findings and offer the possibility of producing inhibitors with different properties. These new data on HIV proteinase specificity illustrate the usefulness of random libraries in the genetic screening system. This approach can be applied to examine any proteinase that has a recognition site extending across several amino acids. PMID- 18384739 TI - Detection of Staphylococcus enterotoxin B using fluorescent immunoliposomes as label for immunochromatographic testing. AB - Staphylococcus enterotoxin B (SEB) is one of several toxins produced by the gram positive bacterium Staphylococcus aureus. SEB is a major cause of food poisoning and represents a significant biological threat with regard to bioterrorism. A rapid, sensitive, and specific method is required to monitor food and water in cases of both natural and intentional contamination by this toxin. This report presents an improved immunochromatographic test (ICT) using immunoliposomes as label for the detection of SEB. For the first time in an ICT, the signal generated by the sulforhodamine B encapsulated into immunoliposomes was measured by fluorescence, allowing a 15-fold increase in sensitivity compared with that for visual detection of colored labels. The ICT was completed within 30 min, providing a limit of detection close to 20 pg/ml in buffer and showing no cross reactivity with the other major toxin of the bacterium, Staphylococcus enterotoxin A. This sensitivity was retained when analyzing SEB spiked in various alimentary matrices, mimicking contaminated foods. Due to the use of fluorescent immunoliposomes as label, the present assay offers the inherent simplicity and speed of a dipstick assay while providing detection of low levels of SEB in real samples. PMID- 18384740 TI - Measuring binding kinetics of surface-bound molecules using the surface plasmon resonance technique. AB - Surface plasmon resonance (SPR) technology and the Biacore biosensor have been widely used to measure the kinetics of biomolecular interactions in the fluid phase. In the past decade, the assay was further extended to measure reaction kinetics when two counterpart molecules are anchored on apposed surfaces. However, the cell binding kinetics has not been well quantified. Here we report development of a cellular kinetic model, combined with experimental procedures for cell binding kinetic measurements, to predict kinetic rates per cell. Human red blood cells coated with bovine serum albumin and anti-BSA monoclonal antibodies (mAbs) immobilized on the chip were used to conduct the measurements. Sensorgrams for BSA-coated RBC binding onto and debinding from the anti-BSA mAb immobilized chip were obtained using a commercial Biacore 3000 biosensor, and analyzed with the cellular kinetic model developed. Not only did the model fit the data well, but it also predicted cellular on and off-rates as well as binding affinities from curve fitting. The dependence of flow duration, flow rate, and site density of BSA on binding kinetics was tested systematically, which further validated the feasibility and reliability of the new approach. PMID- 18384741 TI - Selectivity of bacterial proteome fractionation based on differential solubility: a mass spectrometry evaluation. AB - We investigate the selectivity achieved after differential solubilization of bacterial proteomes following two procedures, both based on successive extraction of proteins in solutions of increasing solubilizing power. Recently, these procedures have gained notable popularity and several commercial kits are now available. A total of 225 proteins in one case and 227 proteins in the other were identified by LC MSMS analysis; 146 of them were identified in both procedures. The proportions of proteins identified as present in only one fraction were 64 and 57%, respectively. The distribution of cytosolic, membrane, and ribosomal proteins among the successive extracts was analyzed in detail. The effect of (1) replacement of low-speed with high-speed centrifugation, (2) omission of detergents in urea solutions, (3) successive washes of pellets, and (4) reproducibility was evaluated. Proteins with positive grand averages of hydropathicity values and membrane proteins were found in all fractions. This study highlights the benefits and limitations of differential solubilization methods, focusing on practical aspects that may strongly influence their selectivity. PMID- 18384742 TI - Ethylene signaling: identification of a putative ETR1-AHP1 phosphorelay complex by fluorescence spectroscopy. AB - The plant ethylene receptor ETR1, which shows substantial sequence homology to typical bacterial histidine kinases, is involved in the coordination of several growth and development processes. Fluorescence polarization studies presented here demonstrate a specific interaction of ETR1 with the histidine-containing transfer protein AHP1, supporting the idea that a phosphorelay module is involved in ethylene signaling. The sensitive assay employed in our studies allows analysis of protein-protein interactions in a homogenous aqueous environment, exact control of external parameters, and quantitative analysis of the affinity and stability of the complex. Thereby it provides the basics for a more quantitative elucidation of phosphorelay modules acquired in phytohormone signaling. PMID- 18384743 TI - Electrostimulation induces cardiomyocyte predifferentiation of fibroblasts. AB - Stem-cell therapy has become a promising therapeutic tool for myocardial repair. Cardiac pre-committed cells, which complete their differentiation in the myocardium, may reduce fibrosis and restore muscle function. However, many questions concerning a precise, functional integration of injected cells remain unanswered. Fibroblasts regulate the cardiac extracellular matrix and are the most abundant cell population in an infarcted area. Electrostimulation is a well known trophic factor and can induce phenotypic changes in myoblasts. The objective of this study was to evaluate the effectiveness of electrical stimulation to induce pre-commitment of fibroblasts into cardiomyocytes in vitro. Using short-time electrostimulation in a cytokine-free culture system, we induced pre-commitment of two fibroblast cell lines to a cardiomyocyte phenotype. This partial differentiation in vitro may facilitate further differentiation within the cardiac environment and result in better electro-mechanical integration of the therapeutically introduced cells. PMID- 18384744 TI - TNF-induced oscillations in combinatorial transcription factor binding. AB - We have shown in two accompanying papers that TNF induces oscillations in (1) approximately 13% of the genome, and (2) the activation of MAP kinase and NF kappaB signaling pathways. Here we aim to bridge oscillations in signal transduction activation to oscillations in genetic output. Specifically, we sought to study how these oscillations can combine in a ligand-specific manner at the level of the promoter to initiate gene transcription. We utilize the late onset gene CD38 as a model gene since it has previously been shown that TNF, but not the related cytokine RANK-L, induces its expression. We find that TNF-induced oscillations in p65 and p50 recruitment to the CD38 promoter correlated with recruitment of MAPK-induced AP-1 recruitment, as analyzed by quantitative ChIP analysis. Through re-ChIP analysis we show that a unique transcriptional complex is seen on the promoter at 3h post-TNF addition, corresponding to the onset of CD38 transcription, which is not seen in the basal state. Moreover, we show that RANK-L was unable to combinatorially recruit AP-1 and NF-kappaB transcription factors to the CD38 promoter, despite inducing the activation of both signaling pathways. These results, in sum with the two accompanying papers, constitute a new paradigm through which cells dynamically orchestrate signaling molecules to coordinate time-resolved gene transcription by the formation of novel time specific transcriptional complexes. PMID- 18384745 TI - An ENU-induced mutation in the lymphotoxin alpha gene impairs organogenesis of lymphoid tissues in C57BL/6 mice. AB - The TNF family is critical for development of lymphoid organs and plays significant roles in regulation of innate and adoptive immune responses. Here, we describe a novel N-ethyl-N-nitrosourea (ENU)-induced mutant strain, HLB382, with a point mutation in the Lta gene, which encodes lymphotoxin (LT) alpha, a member of the TNF family. Mutant mice had no lymph nodes and no Peyer's patches. Microscopically, the spleens had disordered follicles and no germinal centers or discernible marginal zones (MZ). While the development of T cells and follicular B cells was normal, the numbers of NK and MZ B cells were significantly reduced. Interestingly, the numbers of peritoneal B1b cells were significantly increased. Genomic DNA sequences revealed a single base pair insertion in the coding region of Lta resulting in a frame shift and a premature stop codon. This new strain provides opportunities for understanding the full range of Lta gene function on a pure C57BL/6 background. PMID- 18384746 TI - TNF-induced gene expression oscillates in time. AB - With only few exceptions that include Hes-1 p53, and IkappaB, the expression of genes has never been shown to be oscillatory. Here, we show that the inflammatory cytokine TNF triggers oscillations in >5000 genes. We utilize microarrays at 30 min intervals to analyze the pattern of global gene expression in murine macrophages. We find that 15% of genes in the genome underwent a significant >3 fold increase in expression, with 89% of these displaying oscillations at frequencies as low as every 50min. We analyze further two sub-clusters of genes that either began oscillating early or after a lag phase. Through the use of quantitative PCR, we confirm the oscillations and show that the oscillations are continuous. Moreover, we show that these continuous oscillations are not unique to TNF, but that related cytokines such as RANK-L produces oscillations with a unique induction profile. In the two papers accompanying this one, we analyze the mechanism of these oscillations and find that TNF also triggers oscillations in the phosphorylation of MAP kinases, and that these oscillations combine to recruit transcription factors to promoters in a cyclical fashion. The results presented here suggest that gene transcription is a highly dynamic processes, with thousands of genes displaying rapid (<60min) oscillations over time. Considering this dynamism, time-resolved measurements of gene transcription should become the experimental norm. PMID- 18384747 TI - The enzymatic activity of sialidase Neu2 is inversely regulated during in vitro myoblast hypertrophy and atrophy. AB - Sialidase Neu2 is an exoglycosidase that removes terminal sialic acids from glycolipids and glycoproteins. In this study, we investigated Neu2 expression during muscle hypertrophy and atrophy. Neu2 mRNA and enzymatic activity were significantly increased in hypertrophic myofibers. A rise in Neu2 activity was observed after constitutive activation of AKT or Igf-1 treatment as well as in myoblasts treated with vasopressin or trichostatin, an inhibitor of histone deacetylases. In contrast, myofiber atrophy obtained by dexamethasone treatment or starvation triggered a significant loss of Neu2 activity and was paralleled by downregulation of Neu2 transcript levels. Overall, we may conclude that Neu2 enzymatic activity is causally linked to proper muscle differentiation and growth. PMID- 18384748 TI - GABA and synaptic inhibition of mouse cerebellum lacking glutamate decarboxylase 67. AB - gamma-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter and also presumed to be a neurotrophic factor. GABA is synthesized by glutamate decarboxylase (GAD). A mouse lacking a 67kDa isoform of GAD (GAD67) has a reduced GABA level in its brain at birth and does not survive postnatally because of cleft palate. In this study, to investigate the functional and developmental roles of GABA in the postnatal cerebellum, selective GAD67 deletion was achieved using a Cre-loxP strategy. In this mouse, GABA level was reduced to 16-44% in the cerebellum but not in the cerebrum. Inhibitory synaptic transmission to Purkinje cells was seriously impaired. However, the morphology of Purkinje cells and the density of synaptic terminals in the cerebellar cortex appeared unaffected, suggesting that GABA does not participate in cerebellar development substantially. PMID- 18384749 TI - Differential phosphorylation of Cdc25C phosphatase in mitosis. AB - Cdc25 dual-specificity phosphatases coordinate entry into mitosis through activating dephosphorylation of the Mitosis-Promoting Factor, Cdk1-cyclin B1. Activation of Cdc25C at the G2/M transition, involves its dissociation from 14-3 3, together with its hyperphosphorylation on several sites within its regulatory N-terminal domain, mediated by cyclin-dependent kinases and Plk1. Growing evidence suggests that phosphorylation intermediates are likely to precede complete hyperphosphorylation of Cdc25C. To address whether such variants occur in mitotic cells, we raised antibodies directed against different mitotic phosphorylation sites of human Cdc25C, and characterized the phosphorylated species detectable in HeLa cells. In the present study, we provide first-time evidence for the existence of multiple species of Cdc25C in mitotic cell extracts, including full-length and splice variants with different phosphorylation patterns, thereby revealing an intricate network of Cdc25C phosphatases, likely to have distinct biological functions. PMID- 18384750 TI - Sumoylation of Smad3 stimulates its nuclear export during PIASy-mediated suppression of TGF-beta signaling. AB - Sma- and MAD-related protein 3 (Smad3) plays crucial roles in the transforming growth factor-beta (TGF-beta)-mediated signaling pathway, which produce a variety of cellular responses, including cell proliferation and differentiation. In our previous study, we demonstrated that protein inhibitor of activated STATy (PIASy) suppresses TGF-beta signaling by interacting with and sumoylating Smad3. In the present study, we examined the molecular mechanisms of Smad3 sumoylation during PIASy-mediated suppression of TGF-beta signaling. We found that small-interfering RNA-mediated reduction of endogenous PIASy expression enhanced TGF-beta-induced gene expression. Importantly, coexpression of Smad3 with PIASy and SUMO1 affected the DNA-binding activity of Smad3. Furthermore, coexpression of Smad3 with PIASy and SUMO1 stimulated the nuclear export of Smad3. Finally, fluorescence resonance energy transfer analyses revealed that Smad3 interacted with SUMO1 in the cytoplasm. These results suggest that PIASy regulates TGF-beta/Smad3-mediated signaling by stimulating sumoylation and nuclear export of Smad3. PMID- 18384751 TI - TNF-induced MAP kinase activation oscillates in time. AB - Oscillations in the activation of multiple signaling pathways have never been shown before. Our results presented in the previous accompanying paper showed that TNF induces highly dynamic oscillations in mRNA production in approximately 13% of the mouse genome. Here, we further analyze the TNF time-series microarray data and find that multiple signaling components downstream of the TNF receptor undergo oscillations. Prior studies implicate IkappaBalpha and A-20 as the only oscillatory components in the TNF signaling cascade. We find however, that other components, such as TRAF1, displayed oscillations. This suggested the possibility that all signaling output from the TNF receptor may be oscillatory in nature. Indeed, we show that TNF triggers oscillations in the phosphorylation of three MAP kinases, as well as p65. Because Baltimore and colleagues had proposed that NF-kappaB drives the oscillatory nature of the IkappaBalpha/NF-kappaB feedback loop, we studied the effects of an NF-kappaB super-repressor on oscillations in MAPK phosphorylation; we find that the super-repressor altered the amplitude and frequency of MAP kinase phosphorylation, but failed to abolish oscillations. These results attest to a role for NF-kappaB as a modulator, but not the sole determinant of cyclical activation of signal transduction pathways. These results, together with those of the two accompanying papers, constitute a new paradigm through which cells orchestrate signaling molecules to produce highly dynamic physiological processes such as gene transcription and protein secretion. In view of the discovery that multiple phosphorylation pathways display dynamic oscillations, time-resolved, instead of static, measurements of kinase phosphorylation should become the experimental norm. PMID- 18384752 TI - Knowledge sharing and expertise coordination of event response in organizations. AB - This paper provides an overview of opportunities and challenges for expert coordination, knowledge sharing, and task performance using advanced information and communication technologies. Evolving in part from [Hendrick, H., 1991. Ergonomics in organizational design and management. Ergonomics 34(6), 743-756] discussion of macroergonomics, this paper describes the author's framework for systems engineering analysis of information flow and performance at team and organizational units of analysis. Work in the author's research lab has focused on several aspects of information technology use and team interactions to support shared understandings, task demands, and effective responses in responses to events. Multiple empirical studies are summarized describing evaluations of technology use, task cycles and expert knowledge coordination in several settings, including aerospace, healthcare, and project management. PMID- 18384753 TI - Global inequality, and the challenge for ergonomics to take a more dynamic role to redress the situation. AB - This paper argues that ergonomics is more sorely needed, easier to implement, and potentially far more effective in industrially developing countries (IDCs) than where its efforts are presently most concentrated in the less populated, more affluent, technologically advanced world. The reasoning is a simple extension of the principle of diminishing returns in which the further from optimal a situation is, the greater the beneficial effect of any implemented improvement. The paper draws attention to the gap between 'have' and 'have not' cultures, plus the necessity for, and relative ease with which a sustainable ergonomics ethos can be engendered in IDCs. This requires a need to consider network causality, investigating both micro-problems (basic interaction between task and worker) and macro-conditions of the overall scenario (including managerial organisation, planning and responsibility). The two-pronged symbiosis of micro- and macro ergonomics intervention has the potential to achieve both effective and sustainable development within small, medium and large enterprises. PMID- 18384754 TI - Sleep homeostasis: a role for adenosine in humans? AB - Sleep is not the mere absence of wakefulness, but an active state which is finely regulated. The homeostatic facet of sleep-wake regulation is keeping track of changes in 'sleep propensity' (or 'sleep need'), which increases during wakefulness and decreases during sleep. Increased sleep propensity following extended prior wakefulness (sleep deprivation) is counteracted by prolonged sleep duration, but also by enhanced non-rapid-eye-movement (nonREM) sleep intensity as measured by electroencephalographic (EEG) slow-wave activity (SWA, power within approximately 1-4 Hz). This highly reliable regulatory feature of nonREM sleep may be the most important aspect of sleep in relation to its function. The neurochemical mechanisms underlying nonREM sleep homeostasis are poorly understood. Here we provide compelling and convergent evidence that adenosinergic neurotransmission plays a role in nonREM sleep homeostasis in humans. Specifically, a functional polymorphism in the adenosine metabolizing enzyme, adenosine deaminase, contributes to the high inter-individual variability in deep slow-wave sleep duration and intensity. Moreover, the adenosine receptor antagonist, caffeine, potently attenuates the EEG markers of nonREM sleep homeostasis during sleep, as well as during wakefulness. Finally, adenosinergic mechanisms modulate individual vulnerability to the detrimental effects of sleep deprivation on neurobehavioral performance, and EEG indices of disturbed sleep after caffeine consumption. While these convergent findings strongly support an important contribution of adenosine and adenosine receptors to nonREM sleep homeostasis, further research is needed to elucidate the underlying mechanisms that mediate the actions of adenosine on sleep and the sleep EEG. PMID- 18384755 TI - A facile method to screen inhibitors of protein-protein interactions including MDM2-p53 displayed on T7 phage. AB - Protein-protein interactions are essential in many biological processes including cell cycle and apoptosis. It is currently of great medical interest to inhibit specific protein-protein interactions in order to treat a variety of disease states. Here, we describe a facile multiwell plate assay method using T7 phage display to screen for candidate inhibitors of protein-protein interactions. Because T7 phage display is an effective method for detecting protein-protein interactions, we aimed to utilize this technique to screen for small-molecule inhibitors that disrupt these types of interaction. We used the well characterized interaction between p53 and MDM2 and an inhibitor of this interaction, nutlin 3, as a model system to establish a new screening method. Phage particles displaying p53 interacted with GST-MDM2 immobilized on 96-well plates, and the interaction was inhibited by nutlin 3. Multiwell plate assay was then performed using a natural product library, which identified dehydroaltenusin as a candidate inhibitor of the p53-MDM2 interaction. We discuss the potential applications of this novel T7 phage display methodology, which we propose to call 'reverse phage display'. PMID- 18384756 TI - Plasmin is involved in inflammation via protease-activated receptor-1 activation in human dental pulp. AB - Plasmin is a proteolytic enzyme produced from plasminogen by plasminogen activators. We investigated the function of plasmin in human dental pulp fibroblast-like cells. Plasmin induced an increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)) in a concentration-dependent manner. Expression of mRNA for protease-activated receptor-1 (PAR-1) was detected, and the PAR-1 activating peptide SFLLRN induced an increase in [Ca(2+)](i) in the cells. The plasmin-induced increase in [Ca(2+)](i) was inhibited in the presence of the PAR 1 antagonist SCH79797. Plasmin stimulated the expression of interleukin-8 (IL-8) mRNA and prostaglandin E(2) release, which are involved in inflammation. These effects of plasmin on expression of IL-8 mRNA and prostaglandin E(2) release were inhibited in the presence of the PAR-1 antagonist SCH79797. These results suggest that plasmin activates PAR-1 and is involved in inflammation in human dental pulp. PMID- 18384757 TI - Reactions of isodimethoate with human red cell acetylcholinesterase. AB - Isodimethoate is a thermal decomposition product that is present in usual pesticide formulations of dimethoate. Owing to its PO structure the compound is a direct anticholinesterase agent whose properties, to the best of our knowledge, are presented here for the first time. Isodimethoate shows an inhibition rate constant towards human red blood cell acetylcholinesterase (AChE) of 2.3x10(3) M( 1) min(-1) (pH 7.4, 37 degrees C), indicating a somewhat higher potency than found with omethoate, the CYP450-mediated active metabolite of pure dimethoate. Isodimethoate-inhibited AChE shows fast spontaneous reactivation and aging kinetics (half-life 2.3 and 25 min, respectively). The inhibited, non-aged enzyme is readily reactivated by obidoxime (k(r)=9 min(-1); K(D)=0.1 mM) but hardly by pralidoxime at therapeutic concentrations. Interestingly, isodimethoate hydrolyzes readily in buffered solutions at pH 7.4 and 37 degrees C with liberation of methylmercaptan (half-life 16 min). Liberation of N (methyl)mercaptoacetamide, the expected leaving group, was not observed. These properties make isodimethoate a hit-and-run agent that renders part of AChE non reactivatable within a short period of time. The clinical consequences of exposure to or intentional ingestion of isodimethoate-containing dimethoate formulations are a partly untractable AChE shortly after incorporation. In fact, aging of AChE in dimethoate-poisoned patients on admission was much more advanced than expected from the reaction with omethoate. Manufacturers, researching scientists and clinical toxicologists should be aware of this problem. PMID- 18384758 TI - Molecular cloning and pharmacological characterization of rat melatonin MT1 and MT2 receptors. AB - In order to interpret the effects of melatonin ligands in rats, we need to determine their activity at the receptor subtype level in the corresponding species. Thus, the rat melatonin rMT(1) receptor was cloned using DNA fragments for exon 1 and 2 amplified from rat genomic DNA followed by screening of a rat genomic library for the full length exon sequences. The rat rMT(2) receptor subtype was cloned in a similar manner with the exception of exon 1 which was identified by screening a rat genomic library with exon 1 of the human hMT(2) receptor. The coding region of these receptors translates proteins of 353 and 364 amino acids, respectively, for rMT(1) and rMT(2). A 55% homology was observed between both rat isoforms. The entire contiguous rat MT(1) and MT(2) receptor coding sequences were cloned, stably expressed in CHO cells and characterized in binding assay using 2-[(125)I]-Iodomelatonin. The dissociation constants (K(d)) for rMT(1) and rMT(2) were 42 and 130 pM, respectively. Chemically diverse compounds previously characterized at human MT(1) and MT(2) receptors were evaluated at rMT(1) and rMT(2) receptors, for their binding affinity and functionality in [(35)S]-GTPgammaS binding assay. Some, but not all, compounds shared a similar binding affinity and functionality at both rat and human corresponding subtypes. A different pharmacological profile of the MT(1) subtype has also been observed previously between human and ovine species. These in vitro results obtained with the rat melatonin receptors are thus of importance to understand the physiological roles of each subtype in animal models. PMID- 18384759 TI - Treatment of traumatic brain injury in mice with marrow stromal cells. AB - This study was designed to investigate the potential beneficial effects of bone marrow stromal cell (MSC) treatment of traumatic brain injury (TBI) in mice. Twelve female C57BL/6J mice (weight, 21-26 g) were injured with controlled cortical impact and divided into 2 groups (n=6 each). The experimental group was injected with MSCs (0.3x10(6)) intravenously one day after TBI, whereas the control group was injected with saline. MSCs were harvested from male mice, and male to female transplantation was performed to identify male donor cells within female recipient animals. This was achieved by localizing Y chromosomes within the female mice. Neurological function was assessed using the Morris water maze and foot fault tests. All mice were sacrificed 35 days after TBI. Brain sections were stained using in situ hybridization and immunohistochemistry to identify MSCs as well as to analyze vascular density following MSC treatment. Both modalities of testing demonstrated significant improvement in neurological function in the MSC-treated group compared to the saline-treated control group (p<0.05). Histologically, Y chromosome labeled MSCs were easily identified in the injured brain, localized primarily around the lesion boundary zone. There was also a significant increase in vascular density in the lesion boundary zone and hippocampus of MSC-treated mice compared to control mice. This is the first study to show beneficial effects of MSC treatment after TBI in mice. PMID- 18384760 TI - Pretreatment of microcrystalline cellulose flakes with CaCl2 increases the surface area, and thus improves enzymatic saccharification. AB - Glucose production from cellulose flakes with cellulases was improved after pretreatment with saturated CaCl2 at room temperature. When pretreated microcrystalline cellulose flakes (Funacel II, Funakoshi Co., Ltd, Tokyo, Japan) were saccharified with the cellulases, 76.8% of the substrate was converted into glucose within 5 h, whereas the corresponding conversion rate of water-pretreated cellulose flakes was 33.8%. To clarify the mechanism of the promotion, cellobiohydrolase I purified from Trichoderma longibrachiatum was used as the model cellulase, which degraded CaCl2-pretreated cellulose more quickly than the water-pretreated cellulose under tested conditions. The maximum amount of the enzyme bound to CaCl2-pretreated cellulose at 37 degrees C was estimated as 1.14 nmol/mg of cellulose, whereas that to water-pretreated cellulose was 0.527 nmol/mg of cellulose. The specific activity of the bound enzyme greatly decreased with the increase of the surface density (rho) of the bound enzyme, and no significant positive effects of the CaCl2-pretreatment on the specific activity could be observed at the same rho value, suggesting that the promotion was attributed mainly to the increase of the surface area of cellulose. The effect was also observed with dewaxed cotton or filter paper, but not with nata de coco cellulose or bagasse cellulose as the substrates. This suggests that the CaCl2 pretreatment serves to increase the surface area of cellulose flakes via liberation of cellulose particles which were artificially aggregated during harsh drying processes of the flakes. PMID- 18384761 TI - NMR spectroscopic study of cyclodextrin inclusion complexes with A-007 prodrugs. AB - One- and two-dimensional NMR spectroscopy was used to demonstrate the formation of inclusion cyclodextrin complexes with several A-007 prodrugs. These complexes are comprised from the encapsulation of the two phenol moieties of the A-007 prodrugs within the cyclodextrin cavity. Considering the size of the two phenol moieties of the A-007 prodrugs compared to the sizes of alpha-, beta-, and gamma cyclodextrin cavities, we observed complementary binding of the A-007 prodrug with only beta-cyclodextrin, which was also demonstrated spectroscopically. The beta-cyclodextrin inclusion complexes increased the prodrug solubility and modified the prodrug half-life in water. Therefore, beta-cyclodextrin inclusion complexes can be used as an essential form of A-007 prodrug delivery. PMID- 18384762 TI - Synthesis of new 2-phosphono-alpha-D-glycoside derivatives by stereoselective oxa Michael addition to a D-galacto derived enone. AB - The synthesis of new 2-phosphono-alpha-D-glycoside derivatives by stereoselective oxa-Michael addition to an enone derived from D-galactal and containing a phosphonate group is described. Retro-Michael reactions were prevented by tandem acetylation to trap the unstable enolic intermediates. The stereochemistry of the addition products was established by NOESY experiments and explained with molecular mechanics (MM) and density functional theory (DFT) calculations. PMID- 18384763 TI - Inactivation of an invertebrate acetylcholinesterase by sulfhydryl reagents: a reconsideration of the implications for insecticide design. AB - Previously we used site-directed mutagenesis, in vitro expression, and molecular modeling to investigate the inactivation of an invertebrate acetylcholinesterase, cholinesterase 2 from amphioxus, by the sulfhydryl reagents 5,5'-dithiobis(2 nitrobenzoic acid) (DTNB) and N-ethylmaleimide (NEM). We created the mutants C310A, C466A, C310A/C466A and C310A/F312I to assess the roles of the two cysteines and a proposal that the increased rate of inactivation previously found in an F312I mutant was due to increased access of sulfhydryl reagents to Cys310. Our results indicated that both of the cysteines could be involved in inactivation by sulfhydryl reagents, but that the cysteine near the acyl pocket was more accessible. We speculated that the inactivation of aphid AChEs by sulfhydryl reagents was due to the presence of a cysteine homologous to Cys310 and proposed that this residue could be a target for a specific insecticide. Here we reconsider this proposal. PMID- 18384764 TI - The Frizzled Planar Cell Polarity signaling pathway controls Drosophila wing topography. AB - The Drosophila wing is a primary model system for studying the genetic control of epithelial Planar Cell Polarity (PCP). Each wing epithelial cell produces a distally pointing hair under the control of the Frizzled (Fz) PCP signaling pathway. Here, we show that Fz PCP signaling also controls the formation and orientation of ridges on the adult wing membrane. Ridge formation requires hexagonal cell packing, consistent with published data showing that Fz PCP signaling promotes hexagonal packing in developing wing epithelia. In contrast to hair polarity, ridge orientation differs across the wing and is primarily anteroposterior (A-P) in the anterior and proximodistal (P-D) in the posterior. We present evidence that A-P ridge specification is genetically distinct from P-D ridge organization and occurs later in wing development. We propose a two-phase model for PCP specification in the wing. P-D ridges are specified in an Early PCP Phase and both A-P ridges and distally pointing hairs in a Late PCP Phase. Our data suggest that isoforms of the Fz PCP pathway protein Prickle are differentially required for the two PCP Phases, with the Spiny-legs isoform primarily active in the Early PCP Phase and the Prickle isoform in the Late PCP Phase. PMID- 18384765 TI - Time constraints and positional cues in the developing cerebellum regulate Purkinje cell placement in the cortical architecture. AB - To elucidate the mechanisms that regulate neuronal placement and integration in the cerebellar circuitry, we assessed the fate of Purkinje cells transplanted to embryonic, juvenile and adult hosts, asking how architectural changes of the developing cortex influence their anatomical incorporation. Donor Purkinje cells navigate through the host parenchyma either along their natural migratory pathway or following unusual routes. In the latter case, donor neurons fail to orientate correctly and to establish the cortico-nuclear projection. Purkinje cells that follow the physiological route achieve the typical orientation and connectivity, but end displaced in the molecular layer if their arrival in the recipient cortex is delayed. Navigation routes and final settling of donor neurons vary with host age, depending on the ontogenetic construction of cortical layering, and particularly on the maturation of granule cells. The migratory behavior and homing of transplanted Purkinje cells is modified after external granular layer ablation, or neutralization of reelin signaling produced by granule cells. Therefore, although the cerebellar milieu remains receptive for Purkinje cells even after the end of development, correct placement of donor neurons depends on the timing of their migration, related to cerebellar developmental dynamics and granule cell layering. PMID- 18384767 TI - Increased frequency of asynapsis and associated meiotic silencing of heterologous chromatin in the presence of irradiation-induced extra DNA double strand breaks. AB - In meiotic prophase of male placental mammals, the heterologous X and Y chromosomes remain largely unsynapsed, which activates meiotic sex chromosome inactivation (MSCI), leading to formation of the transcriptionally silenced XY body. MSCI is most likely related to meiotic silencing of unsynapsed chromatin (MSUC), a mechanism that can silence autosomal unsynapsed chromatin. However, heterologous synapsis and escape from silencing also occur. In mammalian species, formation of DNA double strand breaks (DSBs) during leptotene precedes meiotic chromosome pairing. These DSBs are essential to achieve full synapsis of homologous chromosomes. We generated 25% extra meiotic DSBs by whole body irradiation of mice. This leads to a significant increase in meiotic recombination frequency. In mice carrying translocation chromosomes with synaptic problems, we observed an approximately 35% increase in asynapsis and MSUC of the nonhomologous region in the smallest chromosome pair following irradiation. However, the same nonhomologous region in the largest chromosome pair, shows complete synapsis and escape from MSUC in almost 100% of the nuclei, irrespective of exposure to irradiation. We propose that prevention of synapsis and associated activation of MSUC is linked to the presence of unrepaired meiotic DSBs in the nonhomologous region. Also, spreading of synaptonemal complex formation from regions of homology may act as an opposing force, and drive heterologous synapsis. PMID- 18384766 TI - CRIP homologues maintain apical cytoskeleton to regulate tubule size in C. elegans. AB - Maintenance of the shape and diameter of biological tubules is a critical task in the development and physiology of all metazoan organisms. We have cloned the exc 9 gene of Caenorhabditis elegans, which regulates the diameter of the single-cell excretory canal tubules. exc-9 encodes a homologue of the highly expressed mammalian intestinal LIM-domain protein CRIP, whose function has not previously been determined. A second well-conserved CRIP homologue functions in multiple valves of C. elegans. EXC-9 shows genetic interactions with other EXC proteins, including the EXC-5 guanine exchange factor that regulates CDC-42 activity. EXC-9 and its nematode homologue act in polarized epithelial cells that must maintain great flexibility at their apical surface; our results suggest that CRIPs function to maintain cytoskeletal flexibility at the apical surface. PMID- 18384768 TI - Exposure to inorganic mercury: from dental amalgam to artisanal gold mining. PMID- 18384769 TI - Cardioprotective effect of fluvastatin on isoproterenol-induced myocardial infarction in rat. AB - The present study was designed to investigate whether fluvastatin, a 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, would attenuate the acute myocardial infarction in isoproterenol-treated rat model via maintaining activities of endogenous antioxidant enzymes. Hemodynamic and electrocardiograph parameters were monitored and recorded continuously, cardiac marker enzymes and antioxidative parameters of plasma and heart tissues were measured, and histopathological examination of heart tissues was performed. Isoproterenol treated rats showed lower of left-ventricular systolic pressure (LVSP), maximum (LVdP/dtmax) and minimum rate of developed left ventricular pressure (LVdP/dtmin), and higher of left ventricular end-diastolic pressure (LVEDP), in addition, a significant rise in ST-segment and increase in content of lactate dehydrogenase, glutamic oxalacetic transaminase, creatine kinase and malondialdehyde, as well as fall in activities of glutathione peroxidase, superoxide dismutase and catalase were observed. Oral administration of fluvastatin (5, 10 and 20 mg/kg, respectively) significantly prevented almost all the parameters of isoproterenol-induced heart failure and myocardial injury that mentioned above. The protective role of fluvastatin on isoproterenol-induced myocardial damage was further confirmed by histopathological examination. There was no significant change in heart rate in all experimental groups. Compared with control group, any indexes in sham rats treated with fluvastatin (20 mg/kg) alone were unaltered (all P>0.05). Our results suggest that fluvastatin has a significant effect on the protection of heart against isoproterenol-induced myocardial infarction through maintaining endogenous antioxidant enzyme activities. PMID- 18384770 TI - Midazolam attenuates the antinociception induced by d-serine or morphine at the supraspinal level in rats. AB - Our recent study has shown that the intracerebroventricular administration of d serine, an endogenous and selective agonist for the glycine site of the N-methyl d-aspartate receptor, alone or in combination with morphine, leads to the potentiation of antinociception on the tail-flick response. Although there is a variety of information concerning the effects of benzodiazepines on opioid induced antinociception, little is known about the effect of benzodiazepines on the N-methyl-d-aspartate receptor agonist-induced antinociception. To clarify the analgesic interactions among the benzodiazepine/GABA(A), N-methyl-d-aspartate and opioid receptors at the supraspinal level, we investigated the effects of intracerebroventricular administration of midazolam, a benzodiazepine receptor agonist, on the antinociception evoked by the intracerebroventricular application of d-serine or morphine. The intracerebroventricular administration of midazolam alone produced hyperalgesia on the tail-flick response in a benzodiazepine receptor antagonist, flumazenil-reversible manner. The antinociception induced by the intracerebroventricular application of d-serine or morphine was attenuated by the intracerebroventricular administration of midazolam. In addition, this inhibitory effect of midazolam on the antinociception of d-serine or morphine was antagonized by the intracerebroventricular administration of flumazenil. Together with the facts that d-serine and midazolam act as selective agonists for the glycine site of the N-methyl-d-aspartate receptor and benzodiazepine/GABA(A) receptor, respectively, these observations suggest a functional interaction between the NMDA and benzodiazepine/GABA(A) receptors in the regulation of antinociception at the supraspinal level. PMID- 18384771 TI - Adiposity, hyperinsulinemia, diabetes and Alzheimer's disease: an epidemiological perspective. AB - The objective of this manuscript is to provide a comprehensive review of the epidemiologic evidence linking the continuum of adiposity, hyperinsulinemia, and diabetes with Alzheimer's disease. The mechanisms for these associations remain to be elucidated, but may include direct actions from insulin, advanced products of glycosilation, cerebrovascular disease, and products of adipose tissue metabolism. Elevated adiposity in middle age is related to a higher risk of Alzheimer's disease. The evidence relating adiposity in old age to Alzheimer's disease is conflicting. Several studies have shown that hyperinsulinemia, a consequence of higher adiposity and insulin resistance, is also related to a higher risk of Alzheimer's disease. Hyperinsulinemia is a risk factor for diabetes, and numerous studies have shown a relation of diabetes with higher Alzheimer's disease risk. Most studies fail the take into account the continuum linking these risk factors which may result in underestimation of their importance in Alzheimer's disease. The implication of these associations is that a large proportion of the world population may be at increased risk of Alzheimer's disease given the trends for increasing prevalence of overweight, obesity, hyperinsulinemia, and diabetes. However, if proven causal, these associations also present a unique opportunity for prevention and treatment of Alzheimer's disease. PMID- 18384773 TI - The systemic inflammatory response after spinal cord injury damages lungs and kidneys. AB - Spinal cord injury (SCI) triggers a well characterized, acute, local inflammation leading to secondary damage at the lesion site. Another little recognized problem may be the activation of circulating inflammatory cells that potentially damage tissues outside the cord. We investigated this problem using severe clip compression SCI in rats. We studied systemic inflammation after SCI and its effects on lungs and kidneys, as dysfunction of these organs is a frequent, early complication after SCI. From 2-24 h after SCI, the number of circulating neutrophils (especially immature cells) significantly increased by 3-10 fold. Flow cytometry experiments revealed that SCI transiently activates these neutrophils, causing increased oxidative responses to phorbolmyristic acid at 2 h after SCI; then, from 4-24 h, the neutrophils were less responsive. Neutrophil longevity was increased (30-50% decrease in apoptosis) at 2-8 h after SCI. Immunohistochemical analyses demonstrated the invasion of neutrophils into lungs and kidneys (2 h-7 d after SCI) and more phagocytic macrophages in lungs (12 h, 3 d after SCI). Myeloperoxidase and matrix metalloproteinase-9 activity in lung and kidney homogenates increased (12 h-7 d after SCI). Expression of COX-2 increased and lipid peroxidation also occurred within this time. Control experiments inducing local cord damage by excitotoxic quisqualate injection verified that SCI per se is sufficient to trigger systemic inflammation and organ damage. In summary, SCI mobilizes and activates neutrophils that then migrate into visceral organs, a phenomenon occurring in parallel with their well-known entry into the cord injury site. The systemic inflammatory response to SCI should be targeted in the development of new therapeutic strategies to treat SCI. PMID- 18384772 TI - Volumetric and ionic regulation during the in vitro development of a corneal endothelial barrier. AB - Corneal endothelium is responsible for generating an ion flux between the corneal stroma and the anterior chamber of the eye that is necessary for the cornea to remain transparent. However, the ion transport regulatory mechanisms that develop during the formation of the endothelial barrier are not known. In this study, we determined the influence of cell confluence on cell volume and intracellular ionic content on the corneal endothelial cells of rabbits. Our results demonstrate that non-confluent endothelial cells display a hypertrophic volume increase, with higher intracellular contents of potassium and chlorine than those of confluent cells. In contrast, when cells reach confluence and the endothelial barrier forms, cell volume decreases and the intracellular contents of potassium and chlorine decrease. Our genetic analysis showed a higher expression of CFTR and CA2 genes in non-confluent cells, and of the gene KCNC3 in confluent cells. These results suggest that the normal ionic current that keeps the corneal stroma dehydrated and transparent is regulated by cell-cell contacts and endothelial cell confluence, and could explain why the loss of corneal endothelial cells is often associated with corneal edema and even blindness. PMID- 18384774 TI - Foreign fetal cells persist in the maternal circulation. AB - OBJECTIVE: To determine whether allogenic fetal cells resulting from donor egg pregnancies persist in maternal circulation. DESIGN: Nested polymerase chain reaction (PCR) amplification of the DYS14 sequence, a region of the Y chromosome, from DNA purified from peripheral blood cells. SETTING: Academic medical center. PATIENT(S): Healthy 18-60-year-old women who have had donor egg pregnancies resulting in a male offspring (n = 11) or, as a control, female offspring (n = 8), at least 1 year previously and without any other source for male cells in their peripheral blood or a healthy male. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Detection of DYS14 sequence by nested PCR. RESULT(S): DYS14 was detected in 5/11 (45%) of women who had donor egg pregnancies resulting in a male offspring, but in 0/8 (0) of women who had donor egg pregnancies resulting in a female offspring. The longest interval between delivery of a male offspring and detection of the DYS14 gene was 9 years. CONCLUSION(S): Unmatched, allogenic fetal cells from donor egg pregnancies are able to persist in the circulation of healthy women for at least 9 years after delivery. This implies a novel mechanism by which immunologic detection is avoided by these cells and may impact on how they may be used for regenerative and transplant medicine. PMID- 18384775 TI - A pilot study of premature ovarian senescence: I. Correlation of triple CGG repeats on the FMR1 gene to ovarian reserve parameters FSH and anti-Mullerian hormone. AB - OBJECTIVE: To assess whether the number of triple CGG expansion of the FMR1 (fragile X) gene, known to correlate at premutation (55-200 repeats) and full mutation (>200 repeats) ranges with risk toward premature ovarian failure (POF), also correlates with milder forms of premature ovarian senescence. DESIGN: Retrospective, controlled cohort study. SETTING: Academically affiliated, private fertility center. PATIENT(S): Forty consecutive, new infertility patients, of which 11 presented with a primary diagnosis of repeated pregnancy loss (controls), 23 with prematurely elevated, age-specific baseline follicle stimulating hormone (FSH) levels (i.e., premature ovarian aging, POA) and 6 with POF. INTERVENTION(S): Determination of number of triple CGG repeats on both alleles of FMR1 gene and of FSH and anti-Mullerian hormone (AMH) levels as a reflection of ovarian reserve. MAIN OUTCOME MEASURE(S): Statistical correlation of higher (allele-2) triple repeat counts with patients' clinical diagnoses and with FSH and AMH levels. RESULT(S): Mean triple CGG counts increased in parallel to increasing severity of premature ovarian senescence. Repeat expansion numbers at all levels correlated statistically to FSH. An AMH level of <1.0 ng/mL statistically correlated to >32 triple repeats. CONCLUSION(S): Over 30 triple CGG repeats denote increased risk (and severity) toward premature ovarian senescence in parallel to increasing expansions. Numbers, considered well within the normal range, therefore already denote risk, suggesting that CGG repeats may represent a new test to predict ovarian function and assess female infertility. PMID- 18384776 TI - CXCL12 enhances exogenous CD4+CD25+ T cell migration and prevents embryo loss in non-obese diabetic mice. AB - OBJECTIVE: To investigate the possible role of CXCL12 in the migration of regulatory T (Treg) cells. DESIGN: Animal model-based study. SETTING: Academic. ANIMAL(S): Pregnant non-obese diabetic (NOD) mice were compared with non immunodeficient mice. INTERVENTION(S): In vivo and in vitro CXCL12 induction. MAIN OUTCOME MEASURE(S): Flow cytometric analysis and Treg cell migratory assay. RESULT(S): A significantly high percentage of spontaneous embryo resorption was observed in both syngeneic and allogeneic pregnant NOD mice. The percentage of embryo loss in allogeneic pregnant NOD mice was significantly decreased by treatment with Treg cells and CXCL12 injection; however, no such effect was observed in syngeneic pregnant NOD mice. In addition, the migration of Treg cells induced by CXCL12 was confirmed by both in vitro and in vivo migratory assays. CXCR4, the specific receptor for CXCL12, was expressed more intensively on Treg cells than on non-Treg CD3(+) T cells, whereas CXCL12 was dominantly expressed in cytokeratin 7(+) trophoblast cells at an early stage of gestation, and its expression reduced gradually during pregnancy. CONCLUSION(S): The higher level of embryo loss in allogeneic pregnant NOD mice may be due to the lack of Treg cells. CXCL12 can cause CXCR4(+) Treg cells to migrate into the pregnant uterus and establish a beneficial microenvironment for the fetus. PMID- 18384777 TI - Altered expression of claudin-3 and claudin-4 in ectopic endometrium of women with endometriosis. AB - OBJECTIVE: To investigate the expression of claudin-3 and claudin-4 in the eutopic and ectopic endometrium of women with endometriosis and to evaluate the role of claudin-3 and claudin-4 in the pathogenesis of endometriosis. DESIGN: Cross-sectional measurement of gene expression levels of claudin-3 and claudin-4 on endometriotic tissue. SETTING: Academic. PATIENT(S): Thirty-five patients with endometriosis and 35 healthy women who were free of endometriosis were recruited for the study. INTERVENTION(S): Expression of claudin-3 and claudin-4 were investigated with immunohistochemical analysis, Western blot, and real-time polymerase chain reaction. Morphologic change of tight junction was also observed in different kinds of endometria. MAIN OUTCOME MEASURE(S): The expression levels of claudin-3 and claudin-4 in epithelial cells from 35 ectopic endometrial tissues, 27 eutopic endometrial tissues from women with endometriosis, and 35 normal endometrial tissues from women without endometriosis. RESULT(S): Expression of claudin-3 and claudin-4 was significantly lower in the ectopic endometriotic tissue than in the eutopic endometrium from women with endometriosis and normal controls at both the messenger RNA and protein levels. No significant difference was found between eutopic endometrium from women with endometriosis and normal endometrium from women without endometriosis. CONCLUSION(S): Down-regulated expression of claudin-3 and claudin-4 in ectopic endometrium suggests that claudin-3 and claudin-4 might play a pathogenic role in the formation of endometriosis. PMID- 18384778 TI - Follicular fluid and serum levels of inhibin A and pregnancy-associated plasma protein A in patients undergoing IVF. AB - OBJECTIVE: To elucidate transport of intrafollicular proteins Inhibin A and pregnancy-associated plasma protein A (PAPP-A) across the follicular fluid (FF)/blood barrier. DESIGN: A retrospective study. SETTING: IVF lab at a university hospital, academic, and industrial research labs. PATIENT(S): Fifty five women undertook the IVF program. INTERVENTION(S): Follicular fluid aspirations and analysis, blood sample drawing, and serum analysis. MAIN OUTCOME MEASURE(S): Concentrations of Inhibin A, PAPP-A, and major serum proteins in FF and serum, total amount of PAPP-A, and Inhibin A in FF. RESULT(S): The FF/blood barrier permeability was calibrated using major serum proteins. The FF/serum ratio decreased with the molecular mass of proteins, and their FF and serum concentrations were well correlated. In contrast, concentrations of Inhibin A in paired serum and FF samples showed a weak correlation (r = 0.563), whereas serum and FF concentrations of PAPP-A were independent of each other. The total amount of Inhibin A in FF correlated well with concentrations of Inhibin A in paired serum samples (r = 0.858), whereas the correlation between the total amount of FF PAPP-A and PAPP-A serum concentrations remains poor (r = 0.215). CONCLUSION(S): These observations suggest that at the day of oocyte retrieval, FF is a major source of serum Inhibin A but not of serum PAPP-A. PMID- 18384779 TI - Impact of a second semen analysis on a treatment decision making in the infertile man with varicocele. AB - OBJECTIVE: To evaluate the impact of second semen analysis (SA) on a treatment decision in infertile men with varicocele and abnormal first SA. DESIGN: Retrospective chart review. SETTING: University hospital. INTERVENTION: None. MAIN OUTCOME MEASURE(S): Standard clinical semen analysis. PATIENT(S): A total of 160 infertile men with varicocele and abnormal first SA were evaluated. Two SA were performed in the same andrology lab 3 to 8 weeks apart. Exclusion criteria were azoospermia, low semen volume, leukocytospermia, febrile illness within 3 months, and habitual heat exposure. RESULT(S): Despite fluctuation in absolute values of sperm concentration, motility, and morphology, the second SA remained abnormal in 111 out of 112 men (99.1%) with abnormal first SA. CONCLUSION(S): Initial abnormal SA is a sufficient indication for varicocele treatment because in most patients the second SA remains abnormal and does not change treatment decision. This approach will expedite treatment of infertile men with varicocele and increase its cost effectiveness. PMID- 18384780 TI - Preterm birth, ovarian endometriomata, and assisted reproduction technologies. AB - OBJECTIVE: To report preterm birth and small for gestational age (SGA) rates from assisted reproduction technologies (ART) patients with ovarian endometriomata compared with control groups. DESIGN: Retrospective cohort study. SETTING: Tertiary university affiliated ART center and Perinatal Data Collection Unit (PDCU). PATIENT(S): Every woman who had an ART singleton baby born between 1991 and 2004 had her database record assessed (N = 4382). Control groups included 1201 singleton babies from ART patients without endometriosis and 2400 randomly selected women from the PDCU database of 850,000 births. INTERVENTION(S): There were 95 singleton ART babies from patients with ovarian endometriomata and 535 ART singleton babies from patients who had endometriosis but no ovarian endometriomata. MAIN OUTCOME MEASURE(S): Preterm birth rates and SGA birth rates. RESULT(S): Preterm birth rate increased only in the ovarian endometriomata group when compared with community birth records (n = 850,000). Furthermore, ART patients with ovarian endometriomata had a statistically significantly increased likelihood of having a SGA baby when compared with other forms of endometriosis. CONCLUSION(S): Rates of preterm birth and SGA babies doubled in infertility patients with ovarian endometriomata who required ART. PMID- 18384781 TI - Expression of the novel gene embryo implantation factor 2 (EMO2) in the mouse uterus at the implantation sites. AB - OBJECTIVE: The aim of this study was to identify a novel implantation-related molecule and to examine EMO2 expression in the mouse uterus during the peri implantation period. DESIGN: Experimental study. SETTING: Research laboratory. ANIMAL(S): Adult ICR mice aged 6-8 weeks. INTERVENTION(S): Adult female mice were mated with fertile males to achieve pregnancy. Implantation was delayed by ovariectomizing pregnant mice on day 4 and administering P during days 5-7; implantation was then initiated by administering E(2). Pseudopregnant mice were obtained by mating females with vasectomized males. MAIN OUTCOME MEASURE(S): The tissue distribution of EMO2 mRNA was detected by reverse transcriptase-polymerase chain reaction, and the uterine expression pattern of the EMO2 protein was determined by immunohistochemistry. RESULT(S): The full cDNA sequence of EMO2 was registered in GenBank (AY372183). EMO2 mRNA expression was observed in all mouse tissues tested. The expression of the EMO2 protein was predominately localized in decidual cells at the implantation site during days 5-6 of pregnancy, and its expression was induced by the active blastocyst and artificially induced decidualization. CONCLUSION(S): Our data indicate that EMO2 may play a key role in the mouse embryo implantation process. PMID- 18384782 TI - Excision of the posterior vaginal fornix is necessary to ensure complete resection of rectovaginal endometriotic nodules of more than 2 cm in size. AB - The minimum distance between the vaginal mucosal epithelium and the endometriotic glands was <1,000 microm in 30 patients (49.2%), <2,000 microm in 44 patients (72.1%), and <5,000 microm in 60 patients (98.4%). Our findings provided histologic evidence that excision of the posterior vaginal fornix was necessary to completely remove large rectovaginal endometriotic nodules. PMID- 18384783 TI - Do stimulation characteristics of the first in vitro fertilization cycle predict pregnancy in women of 40 years old and over? AB - The failure of tests of ovarian reserve to predict clinical pregnancy in women 40 years old and older prompted this study of the predictive value of information obtained during the first cycle of in vitro fertilization (IVF). Stimulation characteristics during the first IVF cycle attempt were unhelpful in predicting the possibility of clinical pregnancy within the first three consecutive IVF cycles. PMID- 18384784 TI - A pilot study of premature ovarian senescence: II. Different genotype and phenotype for genetic and autoimmune etiologies. AB - OBJECTIVE: To assess whether abnormal autoimmune function and number of triple CGG repeats on the FMR1 (fragile X) gene, both historically associated with risk toward premature ovarian senescence, represent independent risk factors. DESIGN: Retrospective cohort study. SETTING: Academically affiliated, private fertility center. PATIENT(S): Forty consecutive, new infertility patients, of which 11 presented with a primary diagnosis of repeat pregnancy loss, 23 with prematurely elevated, age-specific baseline follicle stimulating hormone (FSH) levels (i.e., premature ovarian aging) and 6 with premature ovarian failure. INTERVENTION(S): Determination of triple CGG repeats on both alleles of the FMR1 gene, assessment of ovarian reserve via FSH and anti-Mullerian hormone levels, and evaluation of autoimmune status by antiphospholipid antibody panel, antinuclear antibody panel, total immunoglobulin levels (IgG, IgM, IgA), thyroid antibodies (antiglobulin and antimicrosomal), antiovarian, and antiadrenal antibodies. RESULT(S): Twenty-two of 40 patients (55%) demonstrated autoimmune abnormalities. Women with and without autoimmune abnormalities did not differ in age. Patients with autoimmune abnormalities, however, demonstrated significantly lower FSH levels and higher anti-Mullerian hormone levels. Although triple repeats on the lower count allele (allele-1) of the FMR1 gene did not differ statistically, autoimmune patients demonstrated in the higher count allele (allele-2) significantly fewer triple repeats, significantly fewer triple repeats >or=30, and, in contrast to nonautoimmune patients, a normal mean level of triple repeats. CONCLUSION(S): Abnormal autoimmune function and expansions in triple CGG repeats on the FMR1 gene represent distinctively different etiologies for premature ovarian senescence in infertile patients and may, indeed, constitute its two principal causes. PMID- 18384786 TI - A unique longitudinal muscle contraction pattern associated with transient lower esophageal sphincter relaxation. AB - BACKGROUND & AIMS: Contraction of the longitudinal muscle of the esophagus may play a role in the relaxation and opening of the lower esophageal sphincter (LES). The goal of our study was to determine the pattern and precise temporal correlation between local longitudinal muscle contraction (LMC) of the esophagus during peristalsis and transient LES relaxation (TLESR). METHODS: Esophageal pressures and high-frequency intraluminal ultrasound imaging of the esophagus were recorded in 24 healthy subjects during swallow-induced peristalsis and spontaneous TLESR. Intraluminal multiple impedance recordings were obtained to determine the relationship between "common cavity pressure" and gastroesophageal reflux (GER). RESULTS: During swallow-induced peristalsis, there is simultaneous contraction of circular and longitudinal muscles of the esophagus. On the other hand, TLESR is associated with a distinct pattern of LMC in the esophagus that has the following characteristics: (1) it is restricted to the distal esophagus; (2) it begins before the onset of TLESR and spreads in a retrograde manner; (3) it is generally stronger than the swallow-induced contraction; and (4) it is sustained during the entire duration of TLESR. The increase in esophageal pressure during TLESR is temporally correlated with the contraction of the LM of the distal esophagus, rather than with the impedance recorded GER. CONCLUSIONS: We propose that the LMC of the distal esophagus may play an important role in the relaxation of LES and induction of GER. PMID- 18384785 TI - Screening and surveillance for the early detection of colorectal cancer and adenomatous polyps, 2008: a joint guideline from the American Cancer Society, the US Multi-Society Task Force on Colorectal Cancer, and the American College of Radiology. AB - In the United States, colorectal cancer (CRC) is the third most common cancer diagnosed among men and women and the second leading cause of death from cancer. CRC largely can be prevented by the detection and removal of adenomatous polyps, and survival is significantly better when CRC is diagnosed while still localized. In 2006 to 2007, the American Cancer Society, the US Multi-Society Task Force on Colorectal Cancer, and the American College of Radiology came together to develop consensus guidelines for the detection of adenomatous polyps and CRC in asymptomatic average-risk adults. In this update of each organization's guidelines, screening tests are grouped into those that primarily detect cancer early and those that can detect cancer early and also can detect adenomatous polyps, thus providing a greater potential for prevention through polypectomy. When possible, clinicians should make patients aware of the full range of screening options, but at a minimum they should be prepared to offer patients a choice between a screening test that primarily is effective at early cancer detection and a screening test that is effective at both early cancer detection and cancer prevention through the detection and removal of polyps. It is the strong opinion of these 3 organizations that colon cancer prevention should be the primary goal of screening. PMID- 18384787 TI - Polysialylated NCAM represses E-cadherin-mediated cell-cell adhesion in pancreatic tumor cells. AB - BACKGROUND & AIMS: Inhibition of cell-cell adhesion between epithelial cells represents an early step during tumor metastasis. Down-regulation or perturbation of E-cadherin-mediated adherens junctions is an essential requirement in this process. METHODS: The interaction between polysialylated neural cell adhesion molecule (PSA-NCAM) and the E-cadherin adhesion complex was studied by coimmunoprecipitation assays. The presence of PSA-NCAM was correlated with tumor invasion by using cell-cell aggregation and cell migration assays. The importance of polysialic acid (PSA) in the interaction of NCAM with E-cadherin and inhibition of cell-cell adhesion was confirmed by enzymatic removal of PSA from NCAM and down-regulation of PSA-transferases by siRNA. RESULTS: Expression of oncogenic K-Ras(V12) in pancreatic carcinoma cells resulted in induction of PSA NCAM expression and reduced E-cadherin-mediated cellular adhesion. The association of PSA-NCAM with the E-cadherin adhesion complex correlated with decreased cell-cell aggregation and elevated cell migration of pancreatic carcinoma cells. Enzymatic removal of PSA from NCAM or reduction of polysialyltransferase expression led to reduced association between NCAM and E cadherin and subsequently increased E-cadherin-mediated cell-cell aggregation and reduced cell migration. CONCLUSIONS: Our data suggest the induction of PSA-NCAM by oncogenic K-Ras as a novel molecular mechanism by which E-cadherin-mediated cellular adhesion is reduced and dissemination of tumor cells is facilitated. PMID- 18384788 TI - Combined EUS with FNA and ERCP for the evaluation of patients with obstructive jaundice from presumed pancreatic malignancy. AB - BACKGROUND: An EUS-guided FNA (EUS-FNA) and a therapeutic ERCP are frequently required for the evaluation of patients who were seen for an obstructing periampullary lesion. OBJECTIVE: To determine the feasibility and outcomes of combining an EUS-FNA and a therapeutic ERCP into a single session. DESIGN: Retrospective single-center study. SETTING: Tertiary-referral cancer center. PATIENTS: A total of 114 patients with a suspected malignant obstructing lesion in the pancreatic head. INTERVENTIONS: An EUS with or without FNA plus an ERCP. MAIN OUTCOME MEASUREMENTS: Duration, diagnostic yield, and complication rate of the combined procedures. RESULTS: The mean (SD) total procedure time (EUS, with or without FNA plus ERCP) was 73.6 +/- 30 minutes, with a median of 66 minutes (range 25-148 minutes). In many cases, cytologic diagnosis from FNA became available during an ERCP, which obviated the need for further sampling. EUS-FNA had a sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy of 84.6%, 100%, 100%, 62.9%, and 87.8%, respectively. During an ERCP, endoscopic sphincterotomies were performed in 51 patients, and biliary stents were placed in 96 patients. Twelve patients (10.5%) had a complication, with 6 having postprocedural pancreatitis. LIMITATIONS: Retrospective single-center experience. CONCLUSIONS: Combined EUS-FNA and therapeutic ERCP is technically feasible, with a complication rate no higher than the component procedures, while efficiently providing tissue diagnosis and biliary drainage. PMID- 18384789 TI - Managing anticoagulation and antiplatelet medications in GI endoscopy: a survey comparing the East and the West. AB - BACKGROUND: Anticoagulation and antiplatelet medications may potentiate GI bleeding, and their use may lead to an increased need for a GI endoscopy. We hypothesized that there might be different practice patterns among international endoscopists. OBJECTIVE: To explore the differences in management practices for patients who receive anticoagulation and antiplatelet medications from Eastern and Western endoscopists. DESIGN: International survey study. SETTING: Academic medical centers and private clinics. SUBJECTS: Members of the American Society for Gastrointestinal Endoscopy (ASGE) in Eastern (Korea, Japan, China, India, Thailand, Singapore, Malaysia, and Philippines) and Western (United States and Canada) countries were invited to complete a Web-site-based questionnaire. In addition, the questionnaire was sent to university hospitals in South Korea and academic institutions in the United States. METHODS: A survey was administered that contained detailed questions about the use of an endoscopy in patients on anticoagulation and antiplatelet medications. MAIN OUTCOME MEASUREMENTS: Different opinions and clinical practice patterns regarding the use of anticoagulation and antiplatelet medications by Eastern and Western endoscopists. RESULTS: A total of 105 Eastern and 106 Western endoscopists completed the survey. Western endoscopists experienced more instances of procedure-related bleeding (P = .003) and thromboembolism after withdrawal of medications (P = .016). Eastern endoscopists restarted medications later (1-3 days) than Western endoscopists after a biopsy (same day) (P < .001). Eastern endoscopists withdrew aspirin for more than 7 days before a polypectomy and then restarted it 1 to 3 days after a polypectomy, whereas Western endoscopists performed a polypectomy without withdrawing aspirin (P < .001). ASGE guidelines were followed less often by Eastern than by Western endoscopists (P < .001). LIMITATIONS: Low response rate, heterogeneity of the sample, and recall bias. CONCLUSIONS: The opinions and clinical practice patterns for the management of anticoagulation and antiplatelet medications differed significantly between Eastern and Western endoscopists. The lack of uniformity in practice patterns suggests that more data and better education are required in the area of GI endoscopy for patients on anticoagulation and antiplatelet medications, particularly given that individual patient characteristics may be associated with unique types of complications. PMID- 18384790 TI - Developmental exposure to endocrine disruptor chemicals alters follicular dynamics and steroid levels in Caiman latirostris. AB - Human and wildlife are exposed at critical periods of development to endocrine disruptor chemicals (EDC) that may be responsible for reproductive disorders. To test the hypothesis that in ovum exposure to EDC at a critical period for gonadal organogenesis alters post-hatching folliculogenesis and steroidogenesis in Caiman latirostris, we studied the impact of in ovum exposure to 17 beta-estradiol (E2), bisphenol A (BPA), endosulfan (END) and atrazine (ATZ) on gonadal differentiation, follicular dynamics and circulating levels of steroid hormones in neonatal and juvenile caiman. Since C. latirostris is a species with temperature dependent sex determination, eggs were incubated at male (33 degrees C) or female (30 degrees C) producing temperatures and the effect of EDC was evaluated. Neonatal ovaries exhibited germ cells mainly located in clusters evidencing proliferative activity and type I to III follicles. Juvenile ovaries exhibited germ cells and advanced stages of pre-vitellogenic follicles. Prenatal exposure to the highest doses of E2 (1.4 ppm) or BPA (140 ppm) overrode male temperature effect on sex determination. Neonatal females produced by sex reversion lacked type III follicles, while females prenatally exposed to the lowest doses of E2 (0.014 ppm) and BPA (1.4 ppm) or ATZ (0.2 ppm) showed an increase in type III follicles. Juvenile caiman prenatally exposed to E2 or BPA showed an augmented incidence of multioocyte follicles. Neonatal female caiman exposed in ovum to E2 or BPA had higher estrogen serum levels whereas exposure to E2, BPA, ATZ and END decreased T levels. Present data demonstrates that exposure to EDC during gonadal organogenesis alters follicular dynamics and steroid levels later in life. These effects might have an impact on caiman fertility. PMID- 18384791 TI - Psychiatric morbidity following hysterectomy in Egypt. AB - OBJECTIVE: To evaluate psychiatric morbidity in Egyptian women before and after hysterectomy for benign indications. METHOD: A 2-year prospective observational study at Sohag University Hospital, Egypt, in which 96 women scheduled for hysterectomy were assessed for psychiatric comorbidity before and after the operation using the General Health Questionnaire (GHQ-28), Beck Depression Inventory, and Hamilton Anxiety Scale. RESULTS: Of these, 35 (36.5%) had scores of 4 or higher, signifying psychiatric comorbidity (group 1), and 61 (63.5%) had scores less than 4, suggesting no psychiatric comorbidity (group 2). Postoperatively, severe anxiety and depressive symptoms were common in group 1. In group 2, 48 (78.7%) of the 61 women had scores of 4 or higher. In that subgroup, depressive and anxiety symptoms were more common among nulliparas whereas women with a high parity experienced the least psychiatric comorbidity. CONCLUSIONS: As women undergoing hysterectomy risk psychiatric morbidity, gynecologists should consider less invasive interventions to treat benign conditions. PMID- 18384792 TI - SDF-1alpha/CXCR4 decreases endothelial progenitor cells apoptosis under serum deprivation by PI3K/Akt/eNOS pathway. AB - Recent studies have demonstrated that stromal cell-derived factor-1alpha (SDF 1alpha)/CXCR4 interaction regulates multiple cell signal pathways and a variety of cellular functions such as cell migration, proliferation, survival and angiogenesis. In present study, we aimed to determine the effect of SDF-1alpha on endothelial progenitor cells (EPCs) apoptosis induced by serum deprivation and the implication of phosphoinositide 3-kinase (PI3K)/Akt and mitogen-activated protein kinases (MAPKs) signaling in this effect. EPCs were isolated and characterized. SDF-1alpha decreased EPCs apoptosis induced by serum deprivation in a dose-dependent manner and the inhibitory effect was CXCR4 dependent as confirmed by the total abolishment by AMD3100, a CXCR4-specific peptide antagonist. SDF-1alpha treatment also significant decreased caspase-3 expression and activity. The inhibitory effect of SDF-1alpha on EPCs apoptosis was nearly completely abolished by PI3K inhibitors (either Wortmannin or LY294002) and partially abolished by NOS inhibitor, N(G)-nitro-arginine methyl ester, whereas inhibitors of MAPKs had no significant effect on this inhibitory effect. The treatment of EPCs with SDF-1alpha resulted in time-dependent Akt, eNOS, extracellular-regulated kinase (ERK1/2), p38 MAPK and c-Jun N-terminal kinase (JNK) phosphorylations. These findings suggest that PI3K/Akt/eNOS activation, but not MAPKs activation, is required for the inhibitory effect of SDF-1alpha on EPCs apoptosis. PMID- 18384793 TI - Quantification of age-related shape change of the human rib cage through geometric morphometrics. AB - The aim of this study is to quantify patterns of age-related shape change in the human thorax using Procrustes superimposition. Landmarks (n=106) selected from anonymized computed tomography (CT) scans of 63 adult males free of skeletal pathology were used to describe the form of the rib cage. Multivariate linear regression was used to determine a relationship between landmark location and age. Linear and quadratic models were also investigated. A permutation test employing 1 x 10(5) random trials was used to assess the model significance for both model formulations. Linear relationships between the centroid size (CS) of a landmark set and the corresponding individual's height, weight, and BMI were conducted to enable scaling of the dimensionless results from the Procrustes analysis. A significance level of alpha=0.05 was used for all tests. The average age of the study subjects was 57.0+/-17.3 years. Complete landmark sets were obtained from most of the scans (44 of 63). The quadratic relationship between the age and landmark location was found to be significant (p=0.037), thereby establishing a relationship between the age and thoracic shape change. The linear relationship was mildly significant as well (p=0.073). Significant relationships between the centroid size of the dataset and subject weight, height and BMI were determined, with the best-correlated value being weight (p=0.002, R(2)=0.22). Landmark datasets calculated using the quadratic model exhibited shape change consistent with the clinical observations (increasing kyphosis and rounding of the thoracic cage). Procrustes superimposition represents a potential improvement in the approach used to generate computational models for injury biomechanics studies. The coefficients from the quadratic model are provided and can be used to generate the complete set of model landmark data points at a given age. PMID- 18384794 TI - An improved murine femur fracture device for bone healing studies. AB - Murine models are commonly used to investigate bone healing and test new treatments before human trials. Our objective was to design an improved murine femur fracture device and determine optimal mass and velocity settings for maximal likelihood of transverse fracture. Fracture reproducibility was maximized using an adjustable kinetic energy level, a novel mouse positioning system and an electromagnet striker release assembly. Sixty wild-type mice of 8-12-week-old male and female with a weight of 26.4+/-6.1g were subjected to an experimental postmortem fracture in the left and right femur (n=120) using variable kinetic energy inputs. A best-fit prediction equation for transverse fracture was developed using multivariate linear regression. Transverse fracture was shown to correlate most highly with kinetic energy with a maximum likelihood at mv2=292 where m is mass (g) and v is velocity (m/s). Model validation with a group of 134 anesthetized C57BL/6 mice resulted in a favorable transverse fracture rate of 85.8%. Simple modifications to existing fracture devices can improve accuracy and reproducibility. The results may assist researchers studying the effects of genetic modifications and novel treatments on boney healing in murine femur fracture models. Maintaining kinetic energy parameters within suggested ranges may also aid in ensuring accuracy and reproducibility. PMID- 18384795 TI - Covalent modified hydrophilic polymer brushes onto poly(dimethylsiloxane) microchannel surface for electrophoresis separation of amino acids. AB - A new environmentally friendly method is developed for preventing nonspecific biomolecules from adsorption on poly(dimethylsiloxane) (PDMS) surface via in situ covalent modification. o-[(N-Succinimdyl)succiny]-o'-methyl-poly(ethylene glycol) (NSS-mPEG) was covalently grafted onto PDMS microchannel surface that was pretreated by air-plasma and silanized with 3-aminopropyl-triethoxysilanes (APTES). The modification processes were carried out in aqueous solution without any organic solvent. The mPEG side chains displayed extended structure and created a nonionic hydrophilic polymer brushes layer on PDMS surface, which can effectively prevent the adsorption of biomolecules. The developed method had improved reproducibility of separation and stability of electroosmotic flow (EOF), enhanced hydrophilicity of surface and peak resolution, and decreased adsorption of biomolecules. EOF in the modified microchannel was strongly suppressed, compared with those in the native and silanized PDMS microchips. Seven amino acids have been efficiently separated and successfully detected on the coated PDMS microchip coupled with end-channel amperometric detection. Relative standard deviations (RSDs) of their migration time for run-to-run, day to-day and chip-to-chip, were all below 2.3%. Moreover, the covalent-modified PDMS channels displayed long-term stability for 4 weeks. This novel coating strategy showed promising application in biomolecules separation. PMID- 18384796 TI - No-alignment-strategies for exploring a set of two-way data tables obtained from capillary electrophoresis-mass spectrometry. AB - Hyphenated techniques such as capillary electrophoresis-mass spectrometry (CE-MS) or high-performance liquid chromatography with diode array detection (HPLC-DAD), etc., are known to produce a huge amount of data since each sample is characterized by a two-way data table. In this paper different ways of obtaining sample-related information from a set of such tables are discussed. Working with original data requires alignment techniques due to time shifts caused by unavoidable variations in separation conditions. Other pre-processing techniques have been suggested to facilitate comparison among samples without prior peak alignment, for example, 'binning' and/or 'blurring' the data along the time dimension. All these techniques, however, require optimization of some parameters, and in this paper an alternative parameter-free method is proposed. The individual data tables (X) are represented as Gram matrices (XXT), where the summation is taken over the time dimension. Hence the possible variations in time scale are eliminated, while the time information is at least partly preserved by the correlation structure between the detection channels. For comparison among samples, a similarity matrix is constructed and explored by principal component analysis and hierarchical clustering. The Gram matrix approach was tested and compared to some other methods using 'binned' and 'blurred' data for a data set with CE-MS runs on urine samples. In addition to data exploration by principal component analysis and hierarchical clustering, a discriminant partial least squares model was constructed to discriminate between the samples that were taken with and without the prior intake of a drug. The result showed that the proposed method is at least as good as the others with respect to cluster identification and class prediction. A distinct advantage is that there is no need for parameter optimization, while a potential drawback is the large size of the Gram matrices for data with high mass resolution. PMID- 18384797 TI - Development of a method for the quantification of D-glucaric acid in different varieties of grapefruits by high-performance liquid chromatography and mass spectra. AB - Grapefruits were found to contain D-glucaric acid, which has anticancer properties. In the present investigation, a method has been developed for the quantification of D-glucaric acid in grapefruit by high-performance liquid chromatography (HPLC) using a simple isocratic mobile phase with detection at 210 nm. Grapefruit samples were homogenized, centrifuged and filtered through a 0.45 microm membrane and injected into a HPLC system. The developed method was used for the quantification of D-glucaric acid in nine widely used grapefruit varieties. Furthermore, the identity of D-glucaric acid was confirmed by mass spectra. Seasonal variation of D-glucaric acid within the individual varieties were also measured in fruits harvested during November, February and May. The overall trend of D-glucaric acid level was increased from early to late season fruits. The developed method has a sensitivity of D-glucaric acid as low as 0.05 microg with an accuracy and precision >95%. This method was found to be simple, fast, accurate and reproducible. Moreover, the identity of D-glucaric acid was confirmed by mass spectra. Additionally, the labor intensity and cost of sample preparation were greatly reduced as compared to reported methods. This is the first report on quantification of D-glucaric acid in different varieties of grapefruits from three harvesting sessions. PMID- 18384798 TI - Carbon dioxide modified subcritical water chromatography. AB - A novel method of increasing the elution strength in subcritical water chromatography (SWC) by adding CO2 to the water mobile phase is presented. Since the polarity of water reduces dramatically with increasing temperature, this property is used in SWC to create an isocratic mobile phase with tunable elutropic strength in reversed-phase separations. Unfortunately, thermal stability of the stationary phase dictates the upper temperature limit and therefore also the minimum available mobile phase polarity. As a result SWC is often not very effective at eluting non-polar analytes. However, when CO2 is blended into subcritical water, a considerable reduction in mobile phase polarity results and improves such separations. For example, in conventional SWC 1-octanol is not observed to elute from a PRP-1 column after several hours at the maximum column temperature of 200 degrees C. In contrast to this, when CO2 is present at 180atm (1atm=101325Pa) in the mobile phase, 1-octanol elutes with good peak shape in less than 4min at only 100 degrees C. The technique is applied to the separation of a variety of analytes which have previously been challenging or even not possible to analyze by conventional SWC. Further, the ability to use temperature and composition programming with the blended CO2/water mobile phase in SWC is also presented and discussed. Overall, the developed method considerably extends the range of non-polar analytes amenable to SWC analysis, while maintaining the beneficial conventional SWC features of flame ionization detection and environmental compatibility. PMID- 18384799 TI - Preparation of a TiO2 nanoparticle-deposited capillary column by liquid phase deposition and its application in phosphopeptide analysis. AB - Analysis of phosphopeptides from complex mixtures derived from proteolytic digestion of biological samples is a challenging yet highly important task. Since phosphopeptides are usually present in small amounts, enrichment is often necessary prior to their characterization by mass spectrometry. In this study, a thin layer of titanium dioxide (TiO2) nanoparticles (NPs) was deposited onto the surface of capillary column by liquid phase deposition (LPD) technique and applied to selectively concentrate phosphopeptides from protein digest products. This is, to our knowledge, the first demonstration of using liquid phase deposition to construct in-tube solid phase microextraction devices for biological analysis. By coupling the device off-line or on-line with mass spectrometry analysis, experiments for systematic optimization of loading and washing conditions were carried out, and good trapping selectivity of TiO2 NP deposited capillary columns towards phosphopeptides was demonstrated. PMID- 18384800 TI - A unified classification of stationary phases for packed column supercritical fluid chromatography. AB - The use of supercritical fluids as chromatographic mobile phases allows to obtain rapid separations with high efficiency on packed columns, which could favour the replacement of numerous HPLC methods by supercritical fluid chromatography (SFC) ones. Moreover, despite some unexpected chromatographic behaviours, general retention rules are now well understood, and mainly depend on the nature of the stationary phase. The use of polar stationary phases improves the retention of polar compounds, when C18-bonded silica favours the retention of hydrocarbonaceous compounds. In this sense, reversed-phase and normal-phase chromatography can be achieved in SFC, as in HPLC. However, these two domains are clearly separated in HPLC due to the opposite polarity of the mobile phases used for each method. In SFC, the same mobile phase can be used with both polar and non-polar stationary phases. Consequently, the need for a novel classification of stationary phases in SFC appears, allowing a unification of the classical reversed- and normal-phase domains. In this objective, the paper presents the development of a five-dimensional classification based on retention data for 94 111 solutes, using 28 commercially available columns representative of three major types of stationary phases. This classification diagram is based on a linear solvation energy relationship, on the use of solvation vectors and the calculation of similarity factors between the different chromatographic systems. This classification will be of great help in the choice of the well-suited stationary phase, either in regards of a particular separation or to improve the coupling of columns with complementary properties. PMID- 18384801 TI - Sedimentation of concentrated monodisperse colloidal suspensions: role of collective particle interaction forces. AB - The sedimentation velocities and concentration profiles of low-charge, monodisperse hydroxylate latex particle suspensions were investigated experimentally as a function of the particle concentration to study the effects of the collective particle interactions on suspension stability. We used the Kossel diffraction technique to measure the particle concentration profile and sedimentation rate. We conducted the sedimentation experiments using three different particle sizes. Collective hydrodynamic interactions dominate the particle-particle interactions at particle concentrations up to 6.5 vol%. However, at higher particle concentrations, additional collective particle particle interactions resulting from the self-depletion attraction cause particle aggregation inside the suspension. The collective particle-particle interaction forces play a much more important role when relatively small particles (500 nm in diameter or less) are used. We developed a theoretical model based on the statistical particle dynamics simulation method to examine the role of the collective particle interactions in concentrated suspensions in the colloidal microstructure formation and sedimentation rates. The theoretical results agree with the experimentally-measured values of the settling velocities and concentration profiles. PMID- 18384802 TI - One step preparation of superhydrophobic polymeric surface with polystyrene under ambient atmosphere. AB - Brassica oleracea-like polymer surface is facilely fabricated by one-step casting process using amorphous polystyrene (PS) under ambient atmosphere. The obtained coatings show excellent superhydrophobicity and only possess unitary micro-scale structure, similar to the natural brassica leaf. In addition, a simple topography analysis also roughly verifies superhydrophobic structure of branched and intermingled sticks and bumps. This process provides a fairly easy procedure for preparing superhydrophobic surface from common plastics. Moreover, it demonstrates that the micro/nano-binary structure is not necessary for superhydrophobicity, while unitary micro-scale structure for a polymer surface can exhibit outstanding water repellency as natural lotus. PMID- 18384803 TI - Synergistic effects in flows of mixtures of wormlike micelles and hydroxyethyl celluloses with or without hydrophobic modifications. AB - This work presents experimental results on simple shear and porous media flow of aqueous solutions of two hydroxyethyl celluloses (HEC) and two hydrophobically modified hydroxyethyl celluloses (HMHEC) with different molecular weights. Mixtures of these polymers with a cationic surfactant, cetyltrimethylammonium p toluenesulfonate (CTAT) were also studied. Emphasis was given to the range of surfactant concentrations in which wormlike micelles are formed. The presence of hydrophobic groups, the effect of the molecular weight of the polymers, the surfactant and polymer concentrations, and the effect of the flow field type (simple shear versus porous media flow) were the most important variables studied. The results show that the shear viscosity of HEC/CTAT solutions is higher than the viscosities of surfactant and polymer solutions at the same concentrations, but surface tension measurements indicate that no complex formation occurs between CTAT and HEC. On the other hand, a complex driven by hydrophobic interactions was detected by surface tension measurements between CTAT and HMHEC. In this case, the viscosity of the mixture increases significantly more (up to four orders of magnitude at high CTAT concentrations) in comparison with HEC/CTAT aqueous solutions. Increments in the molecular weight of the polymers increase the interaction with CTAT and the shear viscosity of the solution, but make phase separation more feasible. In porous media flow, the polymer/CTAT mixtures exhibited higher apparent viscosities than in simple shear flows. This result suggests that the extensional component of the flow field in porous media flows leads to a stronger interaction between the polymer and the wormlike micelles, probably as a consequence of change of conformation and growth of the micelles. PMID- 18384804 TI - Laponite-supported titania photocatalysts. AB - This study builds on previous results published on the synthesis and characterization of laponite-supported titania photocatalysts. Titania nanocrystals are prepared prior to addition to the clay dispersion, by a sol-gel synthesis incorporating a microwave hydrothermal step. In addition to previously examinations with XRD, TEM, and FT-IR, the samples are further characterized with SEM, 29Si NMR, and BET N2 sorption to gain additional insight into the effect of TiO2 concentration and surface area on the photoactivity of the samples. PMID- 18384805 TI - Leiomyosarcoma of the pericardium, with epicardial metastases and peripheral eosinophilia in a dog. AB - A dog with a history of dyspnoea, anorexia and ascites showed on examination signs of right-sided heart failure, pleural effusion and peripheral eosinophilia. Diagnostic imaging suggested the presence of a mediastinal mass, and histopathological and immunohistochemical examination of a biopsy sample led to a diagnosis of leiomyosarcoma. On post-mortem examination, an extensive mass was found, which encircled the heart and obliterated the pericardial sac, with probable metastatic spread to the epicardium. Eosinophilic infiltration of the neoplastic mass, lamina propria of the stomach and duodenum, interstitium of the kidney, and submucosa of the bladder was consistent with a possible paraneoplastic eosinophilia. PMID- 18384806 TI - Pathogenic characteristics of the Korean 2002 isolate of foot-and-mouth disease virus serotype O in pigs and cattle. AB - Experimental infection of susceptible cattle and pigs showed that the O/SKR/AS/2002 pig strain of foot-and-mouth disease virus (FMDV) causes an infection that is highly virulent and contagious in pigs but very limited in cattle. Pigs directly inoculated with, or exposed to swine infected with, strain O/SKR/AS/2002 showed typical clinical signs, including gross vesicular lesions in mouth and pedal sites. In addition, FMDV was isolated from, and FMDV genomic RNA was detected in, blood, serum, nasal swabs and oesophageal-pharyngeal (OP) fluid early in the course of infection. Antibodies against the non-structural protein (NSP) 3ABC were detected in both directly inoculated and contact pigs, indicating active virus replication. In contrast, the disease in cattle was atypical. After inoculation, lesions were confined to the infection site. A transient viraemia occurred 1 and 2 days after inoculation, and this was followed by the production of antibodies to NSP 3ABC, indicating subclinical infection. No clinical disease was seen, and no antibodies to NSP 3ABC were present in contact cattle. Additionally, no virus or viral nucleic acid was detected in blood, nasal swab and OP fluid samples from contact cattle. Thus, the virus appeared not to be transmitted from infected cattle to contact cattle. In its behaviour in pigs and cattle, strain O/SKR/AS/2002 resembled the porcinophilic FMDV strain of Cathay origin, O/TAW/97. However, the latter, unlike O/SKR/AS/2002, has reduced ability to grow in bovine-derived cells. The porcinophilic character of O/TAW/97 has been attributed to a deletion in the 3A coding region of the viral genome. However, O/SKR/AS/2002 has an intact 3A coding region. PMID- 18384807 TI - Generation and characterization of a novel anti-rat CD40L antibody with inhibitory activities in vitro and in vivo. AB - The CD40-CD40L interaction plays a critical role in cell-mediated immune responses. Blocking this interaction has been shown to be beneficial in the treatment of various diseases studied in murine models. Although rats are widely used to test therapeutic strategies in several disease models, a monoclonal antibody (mAb) to block the CD40-CD40L interaction in rats is not broadly available. In the present study we generated Armenian hamster fibroblasts expressing rat CD40L and used these to generate a novel anti-rat CD40L mAb (AS1). In vitro studies showed that AS1 was able to block CD40L-induced DC maturation and B cell proliferation. Most importantly, in vivo, AS1 inhibited B cell responses in a dose-dependent fashion, as measured by the production of OVA specific antibodies after subcutaneous immunization with OVA. AS1 was shown to be a powerful tool to modulate Ag presentation in vitro and in vivo. Elucidating the effect of AS1 in various rat models for human diseases will provide more insight into blocking the CD40-CD40L interaction as a therapeutic strategy to prevent human diseases. PMID- 18384808 TI - The kinetic and equilibrium molten globule intermediates of apoleghemoglobin differ in structure. AB - An important question in protein folding is whether molten globule states formed under equilibrium conditions are good structural models for kinetic folding intermediates. The structures of the kinetic and equilibrium intermediates in the folding of the plant globin apoleghemoglobin have been compared at high resolution by quench-flow pH-pulse labeling and interrupted hydrogen/deuterium exchange analyzed in dimethyl sulfoxide. Unlike its well studied homolog apomyoglobin, where the equilibrium and kinetic intermediates are quite similar, there are striking structural differences between the intermediates formed by apoleghemoglobin. In the kinetic intermediate, formed during the burst phase of the quench-flow experiment, protected amides and helical structure are found mainly in the regions corresponding to the G and H helices of the folded protein, and in parts of the E helix and CE loop regions, whereas in the equilibrium intermediate, amide protection and helical structure are seen in parts of the A and B helix regions, as well as in the G and H regions, and the E helix remains largely unfolded. These results suggest that the structure of the molten globule intermediate of apoleghemoglobin is more plastic than that of apomyoglobin, so that it is readily transformed depending on the solution conditions, particularly pH. Thus, in the case of apoleghemoglobin at least, the equilibrium molten globule formed under destabilizing conditions at acid pH is not a good model for the compact intermediate formed during kinetic refolding experiments. Our high precision kinetic analysis also reveals an additional slow phase during the folding of apoleghemoglobin, which is not observed for apomyoglobin. Hydrogen exchange pulse-labeling experiments show that the slow-folding phase is associated with residues in the CE loop, which probably forms non-native structure in the intermediate that must be resolved before folding can proceed to completion. PMID- 18384810 TI - Protein unfolding, and the "tuning in" of reversible intermediate states, in protic ionic liquid media. AB - Protic ionic liquids (PILs) are currently being shown to be as interesting and valuable to chemical manipulations as the well-known aprotic ionic liquids (APIL). PILs have the additional advantage that the proton activity (PA) can be adjusted by the choice of Bronsted base and Bronsted acid used in their formation. In the absence of solvent, the PA plays the role of pH in ordinary solutions. Previously, we have shown that solution of proteins in ionic-liquid rich solutions conveys surprising stabilization against hydrolysis and aggregation, permitting multiple unfold/refold cycles without loss to aggregation. Here, we show that the denaturing temperatures of both hen egg white lysozyme and ribonuclease A are sensitive to the PA of the PIL as much as they are to pH in aqueous solutions. A maximum stability for more basic solutions is found, and the unfolding process is well described by the two-state (cooperative) model. Finally, we show that, by PA tuning, the PILs can select folding pathways featuring the postulated intermediates so that they are fully populated during the unfolding process. The intermediates are themselves capable of multiple unfold/refold cycles with little loss per cycle to aggregation process. PMID- 18384809 TI - Temperature-dependent Hammond behavior in a protein-folding reaction: analysis of transition-state movement and ground-state effects. AB - Characterization of the transition-state ensemble and the nature of the free energy barrier for protein folding are areas of intense activity and some controversy. A key issue that has emerged in recent years is the width of the free-energy barrier and the susceptibility of the transition state to movement. Here we report denaturant-induced and temperature-dependent folding studies of a small mixed alpha-beta protein, the N-terminal domain of L9 (NTL9). The folding of NTL9 was determined using fluorescence-detected stopped-flow fluorescence measurements conducted at seven different temperatures between 11 and 40 degrees C. Plots of the log of the observed first-order rate constant versus denaturant concentration, "chevron plots," displayed the characteristic V shape expected for two-state folding. There was no hint of deviation from linearity even at the lowest denaturant concentrations. The relative position of the transition state, as judged by the Tanford beta parameter, beta(T), shifts towards the native state as the temperature is increased. Analysis of the temperature dependence of the kinetic and equilibrium m values indicates that the effect is due to significant movement of the transition state and also includes a contribution from temperature-dependent ground-state effects. Analysis of the Leffler plots, plots of Delta G versus Delta G degrees, and their cross-interaction parameters confirms the transition-state movement. Since the protein is destabilized at high temperature, the shift represents a temperature-dependent Hammond effect. This provides independent confirmation of a recent theoretical prediction. The magnitude of the temperature-denaturant cross-interaction parameter is larger for NTL9 than has been reported for the few other cases studied. The implications for temperature-dependent studies of protein folding are discussed. PMID- 18384811 TI - Evolution of antiparallel two-domain membrane proteins: tracing multiple gene duplication events in the DUF606 family. AB - X-ray crystallography has revealed that many integral membrane proteins consist of two domains with a similar fold but opposite (antiparallel) orientation in the membrane. The proteins are believed to have evolved by gene duplication and gene fusion events from a dual topology ancestral membrane protein, that adapted both orientations in the membrane and formed antiparallel homodimers. Here, we present a detailed analysis of the DUF606 family of bacterial membrane proteins that contains the entire collection of intermediate states of such an evolutionary pathway: single genes that would code for dual topology homodimeric proteins, paired genes coding for homologous proteins with a fixed but opposite orientation in the membrane that would form heterodimers, and fused genes that encode antiparallel two-domain fusion proteins. Two types of paired genes can be discriminated corresponding to the order in which the genes coding for the two oppositely oriented proteins occur in the operon. On the protein level, the heterodimers resulting from the two types of gene pairs are indistinguishable. In contrast, two types of fused genes corresponding to the two possible orders in which the oppositely oriented domains are present in the encoded proteins, do result in discernible types of proteins. The large number of genetic and protein states in the DUF606 family allowed for a detailed phylogenic analysis that revealed a total of nine independent duplication events in the DUF606 family, five of which resulted in paired genes, and four resulted in fused genes. Noticeably, there was no evidence for a sequential mechanism in which fusions evolve from a pair of genes. Rather, an evolutionary mechanism is proposed by which antiparallel two-domain proteins are the direct result of a gene duplication event. Combining the phylogeny of proteins and hosting microorganisms allowed for a reconstruction of the evolutionary pathway. PMID- 18384812 TI - Synthetic antibody libraries focused towards peptide ligands. AB - Synthetic antibody libraries have proven immensely useful for the de novo isolation of antibodies without the need for animal immunization. Recently, focused libraries designed to recognize particular classes of ligands, such as haptens or proteins, have been employed to facilitate the selection of high affinity antibodies. Focused libraries are built using V regions encoding combinations of canonical structures that resemble the structural features of antibodies that bind the desired class of ligands and sequence diversity is introduced at residues typically involved in recognition. Here we describe the generation and experimental validation of two different single-chain antibody variable fragment libraries that efficiently generate binders to peptides, a class of molecules that has proven to be a difficult target for antibody generation. First, a human anti-peptide library was constructed by diversifying a scaffold: the human variable heavy chain (V(H)) germ line gene 3-23, which was fused to a variant of the human variable light chain (V(L)) germ line gene A27, in which L1 was modified to encode the canonical structure found in anti-peptide antibodies. The sequence diversity was introduced into 3-23 (V(H)) only, targeting for diversification residues commonly found in contact with protein and peptide antigens. Second, a murine library was generated using the antibody 26 10, which was initially isolated based on its affinity to the hapten digoxin, but also binds peptides and exhibits a canonical structure pattern typical of anti peptide antibodies. Diversity was introduced in the V(H) only using the profile of amino acids found at positions that frequently contact peptide antigens. Both libraries yielded binders to two model peptides, angiotensin and neuropeptide Y, following screening by solution phage panning. The mouse library yielded antibodies with affinities below 20 nM to both targets, although only the V(H) had been subjected to diversification. PMID- 18384813 TI - A kinetic model of transcription initiation by RNA polymerase. AB - We establish a sequence-dependent kinetic model for the later stage of transcription initiation by RNA polymerase. We suggest that there are three reaction pathways, the abortive pathway, the scrunching pathway and the escape pathway, competitive with each other at each site during the transcription initiation. Using this three-pathway model, we mainly calculate the maximum sizes of the abortive transcripts, the abortive probabilities and the abortive/productive ratios for different promoters by Monte Carlo simulation and analytical methods. These results are quantitatively comparable with the experimental observations. In particular, our model can account for the unproductive initial transcribing complex and the nucleoside triphosphate concentration dependence of the transcription initiation, which have been found in the experiments and were hardly understood by the previous two-pathway kinetic competition model. PMID- 18384814 TI - miR-21 Gene expression triggered by AP-1 is sustained through a double-negative feedback mechanism. AB - miR-21 has been reported to be highly expressed in various cancers and to be inducible in a human promyelocytic cell line, HL-60, after phorbol 12-myristate 13-acetate (PMA) treatment. To examine molecular mechanisms involved in miR-21 expression, we analyzed the structure of the miR-21 gene by determining its promoter and primary transcripts. We show that activation protein 1 (AP-1) activates the miR-21 transcription in conjugation with the SWI/SNF complex, after PMA stimulation, through the conserved AP-1 and PU.1 binding sites in the promoter identified here. The previous findings of enhanced miR-21 expression in several cancers may therefore reflect the elevated AP-1 activity in these carcinomas. A single precursor RNA containing miR-21 was transcribed just downstream from the TATA box in this promoter, which is located in an intron of a coding gene, TMEM49. More important, expression of this overlapping gene is completely PMA-independent and all its transcripts are polyadenylated before reaching the miR-21 hairpin embedding region, indicating that miRNAs could have their own promoter even if overlapped with other genes. By available algorithms that predict miRNA target using a conservation of sequence complementary to the miRNA seed sequence, we next predicted and confirmed that the NFIB mRNA is a target of miR-21. NFIB protein usually binds the miR-21 promoter in HL-60 cells as a negative regulator and is swept off from the miR-21 promoter during PMA induced macrophage differentiation of HL-60. The translational repression of NFIB mRNA by miR-21 accelerates clearance of NFIB in parallel with the simultaneous miR-21-independent transcriptional repression of NFIB after PMA stimulation. Since exogenous miR-21 expression moderately induced endogenous miR-21, an evolutionarily conserved double-negative feedback regulation would be operating as a mechanism to sustain miR-21 expression. PMID- 18384815 TI - The characteristics of species in an evolutionary food web model. AB - We explore the consequences of modifying the way in which species are defined in an evolutionary food web model. In the original version of the model, the species were defined in terms of a fixed number of features, chosen from a large number of possibilities. These features represented phenotypic and behavioural characteristics of the species. Speciation consisted in occasionally replacing one of the features by another. Here we modify this scheme by firstly allowing for a richer structure and secondly by testing whether we are able to eliminate the need for an explicit choice of features altogether. In the first case we allow for changing the number of features which define a species, as well as their nature, and find that in the resulting webs the higher trophic levels typically contain species with the greatest number of features. In the second case, by a simplification of the mechanisms for inter and intra-species competition, we construct a model without any explicit features and find that we are still able to grow model food webs. We assess the quality of the food webs produced and discuss the consequences of our findings for the future modelling of food webs. PMID- 18384816 TI - Prevention of avian influenza epidemic: what policy should we choose? AB - Human-to-human transmission of the avian influenza has been extremely rarely reported, and is considered as limited, inefficient and unsustained. However, experts warn an occurrence of "mutant avian influenza", which can easily spread among humans, because the avian influenza is already endemic, in particular in Asian poultry, and it is evolving in domestic and wild birds, pigs and humans. Outbreak of such mutant avian influenza in the human world may have devastating consequences, which are comparable with these for the 1918 "Spanish influenza". In this paper we develop a mathematical model for the spread of the mutant avian influenza, and explore the effectivity of the prevention policies, namely the elimination policy which increases the effective additional death rate of the infected birds and the quarantine policy which reduces the number of infective contacts. PMID- 18384817 TI - How much non-coding DNA do eukaryotes require? AB - Despite tremendous advances in the field of genomics, the amount and function of the large non-coding part of the genome in higher organisms remains poorly understood. Here we report an observation, made for 37 fully sequenced eukaryotic genomes, which indicates that eukaryotes require a certain minimum amount of non coding DNA (ncDNA). This minimum increases quadratically with the amount of DNA located in exons. Based on a simple model of the growth of regulatory networks, we derive a theoretical prediction of the required quantity of ncDNA and find it to be in excellent agreement with the data. The amount of additional ncDNA (in basepairs) which eukaryotes require obeys N(DEF)=1/2 (N(C)/N(P)) (N(C)-N(P)), where N(C) is the amount of exonic DNA, and N(P) is a constant of about 10 Mb. This value N(DEF) corresponds to a few percent of the genome in Homo sapiens and other mammals, and up to half the genome in simpler eukaryotes. Thus, our findings confirm that eukaryotic life depends on a substantial fraction of ncDNA and also make a prediction of the size of this fraction, which matches the data closely. PMID- 18384818 TI - Polycyclic musks in green-lipped mussels (Perna viridis) from Hong Kong. AB - Six polycyclic musk compounds [Cashmeran (DPMI), Celestolide (ADBI), Phantolide (AHMI), Traseolide (ATII), Tonalide (AHTN), and Galaxolide (HHCB)] were analysed in marine green-lipped mussels (Perna viridis) from Hong Kong. ADBI, HHCB and AHTN were detected in almost all samples, while AHMI, ATII and DPMI were not detected. Concentrations of ADBI, HHCB and AHTN in mussels ranged from below detection limit-0.0743 (mean: 0.0246), 0.247-6.08 (mean: 1.15) and 0.0591-0.738 (mean: 0.190)mg/kg lipid weight, respectively. Mussels from two sampling sites in central Victoria Harbour contained the highest total polycyclic musk levels, suggesting that these waters are heavily influenced by domestic sewage. Concentrations of HHCB and AHTN detected in the mussel samples were the second highest and the highest levels, respectively, compared to global concentrations. A preliminary risk assessment indicated that HHCB and AHTN in mussels pose little or no threat to the health of shellfish consumers. Nevertheless, more comprehensive studies are required to further assess the ecological and human health risks associated with polycyclic musks. PMID- 18384819 TI - Enhancement of long-term potentiation by brain-derived neurotrophic factor requires adenosine A2A receptor activation by endogenous adenosine. AB - The excitatory action of brain-derived neurotrophic factor (BDNF) on synaptic transmission is triggered by adenosine A2A receptor activation. Since high frequency neuronal firing, such as that inducing long-term potentiation (LTP), favours both A2A receptor activation and BDNF effects on transmission, we now evaluated the influence of adenosine on the facilitatory action of BDNF upon CA1 hippocampal LTP. theta-Burst stimulation of the pyramidal inputs induced a significant and persistent increase in field EPSP slopes, and this potentiation was augmented in the presence of BDNF (20 ng/ml), an action prevented by the inhibitor of Trk receptor autophosphorylation, K252a (200 nM). Removal of endogenous extracellular adenosine with adenosine deaminase (ADA, 1 U/ml), as well as the antagonism of adenosine A2A receptors with SCH58261 (100 nM), prevented the excitatory action of BDNF upon LTP. In an adenosine depleted background (with ADA), activation of adenosine A2A receptors (with 10nM CGS21680) restored the facilitatory effect of BDNF on LTP; this was fully prevented by the protein kinase A inhibitor, H-89 (1 microM) and mimicked by the adenylate cyclase activator, forskolin (10 microM). In similar experiments, activation of adenosine inhibitory A1 receptors (with 5 nM CPA) did not affect the facilitatory effect of BDNF. In conclusion, the facilitatory action of BDNF upon hippocampal LTP is critically dependent on the presence of extracellular adenosine and A2A receptor activation through a cAMP/PKA-dependent mechanism. Since extracellular adenosine accumulates upon high-frequency neuronal firing, the present results reveal a key process to allow the influence of BDNF upon synaptic plasticity. PMID- 18384820 TI - Supersensitivity to mu-opioid receptor-mediated inhibition of the adenylyl cyclase pathway involves pertussis toxin-resistant Galpha protein subunits. AB - Sustained administration of opioids leads to antinociceptive tolerance, while prolonged association of L-type Ca2+ channel blockers (e.g. nimodipine) with opioids results in increased antinociceptive response. Herein, we investigated the changes in mu-opioid receptor signalling underlying this shift from analgesic tolerance to supersensitivity. Thus, the interaction of mu-opioid receptors with G proteins and adenylyl cyclase was examined in lumbar spinal cord segments of rats. In control animals, the mu-opioid selective agonists, sufentanil and DAMGO, stimulated [35S]5'-(gamma-thio)-triphosphate ([35S]GTP gamma S) binding and inhibited forskolin-stimulated adenylyl cyclase activity, through a mechanism involving pertussis toxin (PTX) sensitive G alpha(i/o) subunits. Seven days of chronic sufentanil treatment developed antinociceptive tolerance associated with a reduction in mu-agonist-induced [35S]GTP gamma S binding, mu-agonist-induced adenylyl cyclase inhibition, and co-precipitation of G alpha o, G alpha i2 G alpha z and G alpha q11 subunits with mu-opioid receptors. In contrast, combined nimodipine treatment with sufentanil over the same period increased the sufentanil analgesic response. This antinociceptive supersensitivity was accompanied by a significant increase of mu-agonist-induced inhibition of adenylyl cyclase that was resistant to the antagonism by PTX. In good agreement, co-precipitation of the PTX-resistant, G alpha z and G alpha q/11 subunits with mu-opioid receptors was not lowered. On the other hand, the PTX-sensitive subunits, G alpha i2 and G alpha o, as well as agonist-stimulated [35S]GTP gamma S binding were still reduced. Our results demonstrate that mu-opioid analgesic tolerance follows uncoupling of spinal mu-opioid receptors from their G proteins and linked effector pathways. Conversely, the enhanced analgesic response following combined nimodipine treatment with sufentanil is associated with adenylyl cyclase supersensitivity to the opioid inhibitory effect through a mechanism involving PTX-resistant G protein subunits. PMID- 18384821 TI - Sleep deprivation affects sexual behavior and tyrosine hydroxylase (TH) levels in sexually experienced male rats. AB - Paradoxical sleep deprivation (PSD) produces alterations in dopaminergic systems and also modifies sexual behavior. In this work we evaluated PSD effects on the sexual response and tyrosine hydroxylase (TH) expression in dopaminergic pathways related to sexual behavior of naive and sexual experienced rats. Male Wistar rats had their sexual behavior evaluated in 6 copulatory tests, with a 4 days interval. In these tests, the animals interacted with a receptive female and parameters that compose each component of the male sexual reply (initiation, arousal and ejaculation) were evaluated. After the 5th test, the animals were randomly divided in 2 groups, control and PSD, and 96 h later they were submitted to the last copulatory test. PSD facilitated the excitatory and the ejaculatory component, increasing the copulatory efficiency. In addition, reduced mount frequency and ejaculation latency were observed. The temporal patterning of the sexual behavior was modified, with reduction in the number of mount bouts. PSD per se was not able to modify TH levels, but in PSD sexual trained rats, an increase in the number of TH-immunoreactive cellular bodies in all dopaminergic areas evaluated was detected. Our data suggest that PSD facilitates the sexual response and this facilitation combined to sexual training could be the consequence of increased TH levels in dopaminergic pathways related to sexual reply. PMID- 18384822 TI - Accumulation of apocarotenoids in mycorrhizal roots of leek (Allium porrum). AB - Colonization of the roots of leek (Allium porrum L.) by the arbuscular mycorrhizal fungus Glomus intraradices induced the formation of apocarotenoids, whose accumulation has been studied over a period of 25 weeks. Whereas the increase in the levels of the dominating cyclohexenone derivatives resembles the enhancement of root length colonization, the content of mycorradicin derivatives remains relatively low throughout. Structural analysis of the cyclohexenone derivatives by mass spectrometry and NMR spectroscopy showed that they are mono- and diglycosides of 13-hydroxyblumenol C and blumenol C acylated with 3-hydroxy-3 methyl-glutaric and/or malonic acid. Along with the isolation of three known compounds five others are shown to be hitherto unknown members of the fast growing family of mycorrhiza-induced cyclohexenone conjugates. PMID- 18384823 TI - Exotic myiasis caused by 19 larvae of Cordylobia anthropophaga in Namibia and identified using molecular methods in Japan. AB - A case of exotic myiasis caused by tumbu fly (Cordylobia anthropophaga) parasitism acquired while travelling in the Republic of Namibia is reported. This is the fifth case reported in Japan, and is very unusual in that the patient was infected with 19 larvae. This is also the first case diagnosed using molecular methods in Japan. We cultured the extracted larvae in vitro and successfully obtained pupae. PMID- 18384824 TI - Synthesis and biological evaluation of some new A,B-ring modified steroidal D lactones. AB - Starting from the D-homo lactones of androst-4-en-3-one 3 and 4, prepared from 1 and 2, the new 17a homolactones 5-12, 14 and 15, were synthesized. The 4-hydroxy compounds 9 and 10 were obtained through the reaction of 4alpha,5alpha- (5 and 7) and 4beta,5beta- (6 and 8) epoxides with formic acid. The epoxides 5 and 6 were prepared from compound 3, and epoxides 7 and 8 from compound 4 by oxidation with H(2)O(2) under basic conditions. Compound 1 served as a starting substance for obtaining lactones 11-13. Oxidation of compound 1 with m-chloroperbenzoic acid yielded 11 and 12, but compound 13 gave 14. Compound 15 was obtained from 13 by oxidation with H(2)O(2) under basic conditions. The structures of epoxides 6 and 14 were confirmed by X-ray structural analysis. Cytotoxic activity against three tumor cell lines (human breast adenocarcinoma ER+, MCF-7, human breast adenocarcinoma ER-, MDA-MB-231, and prostate cancer PC3) was evaluated. Compounds 6 and 14 showed strong activity against PC3, the IC(50) being 10.6 and 2.2 microM, respectively, whereas compounds 3 and 8 showed strong activity against MDA-MB-231 (IC(50) is 9.3 and 3.6 microM, respectively). Aromatase inhibition assay showed that the tested compounds 9, 10, and 14 possess lower activity compared to formestane. PMID- 18384825 TI - Progesterone receptor (PROGINS) polymorphism and the risk of ovarian cancer. AB - The present case-control study evaluates the role of the progesterone receptor (PR) polymorphism known as PROGINS as a risk factor for ovarian cancer development and investigates the association between these genetic variants and clinical/pathologic variables of ovarian cancer. PROGINS polymorphism was examined, by polymerase chain reaction, in a total of 80 patients with ovarian cancer and 282 control subjects. The frequencies of PROGINS polymorphism T1/T1, T1/T2, and T2/T2 were 71.3, 15.0 and 13.8% in ovarian cancer patients and 78.37, 21.63 and 0% in controls, respectively. The chi(2)-test showed a higher incidence of the T2/T2 genotype (P=0.001) in the ovarian cancer group. In addition, women carrying a mutated allele (T2) showed approximately 2.2 times higher risk of ovarian cancer development as compared to women who have a variant allele (odds ratio (OR)=2.2; 95% CI=1.80-3.54). Regarding the clinical and pathologic findings observed within the cancer group, there was a significant correlation between PROGINS polymorphism and patients with a familial history (chi(2)=6.776; P=0.009; Fischer exact test, P=0.01). In this regard, patients with familial antecedents have a 4.7 times higher likelihood to have at least one risk allele (T2) as compared with patients without familial antecedents (OR=4.69; 95% CI=1.38-15.87). No correlations were observed among the other variables. These data suggest that the PROGINS polymorphism T2/T2 genotype might be associated with an increased risk of ovarian cancer. PMID- 18384826 TI - Ion trap MS/MS of intact testosterone and epitestosterone conjugates--adducts, fragile ions and the advantages of derivatisation. AB - In ion trap mass spectrometry, fragile ions may fragment under the application of resonance ejection during precursor mass isolation, reducing MS/MS spectral intensity. In this study the steroidal epimers testosterone glucuronide (TG) and epitestosterone glucuronide (EG) have been chosen as a model for exploring whether compound structure is linked to ion trap fragility. Both compounds form multiple adducts by ESI-MS, namely protonation, ammonium and sodium, however, the mass spectrum of EG displays a more intense ammonium adduct peak than TG. [TG+NH(4)](+), [EG+NH(4)](+) and [EG+H](+) were found to be fragile ions. To explain the differences in adduct formation and fragility, molecular modelling was employed. Ammonium adduction was localised to the glucuronide ring oxygens and while EG has eight possible adduction sites, only seven were located for TG explaining the increased ammonium adduct abundance with EG. In EG the bond between the steroid and the glucuronide was slightly longer and the oxygen in this bond was more basic than TG. This shows that the EG bond is weaker which may contribute to the fact that [EG+H](+) but not [TG+H](+) is fragile. To investigate whether stability could be restored by chemical means, EG was derivatised with tris(trimethoxyphenyl)phosphonium chloride or methylated on the carboxylic acid and Girard P or methoxylamine on the 3-keto group. Derivatisation of the steroid rather than the glucuronide eliminated fragility and using a charged derivative eliminated adduct formation. This work demonstrates the importance of carefully considering the nature of the derivative and the site of derivatisation. PMID- 18384827 TI - A novel fitness proxy in structured locally finite metapopulations with diploid genetics, with an application to dispersal evolution. AB - Many studies of evolutionarily stable strategies (ESS) for technical reasons make the simplification that reproduction is clonal. A post-hoc justification is that in the simplest eco-evolutionary models more realistic genetic assumptions, such as haploid sexual or diploid sexual cases, yield results compatible with the clonal ones. For metapopulations the technical reasons were even more poignant thanks to the lack of accessible fitness proxies for the diploid case. However, metapopulations are also precisely the sort of ecological backdrop for which one expect discrepancies between the evolutionary outcomes derived from clonal reproduction and diploid genetics, because substantially many mutant homozygotes appear locally even though the mutant is rare globally. In this paper we devise a fitness proxy applicable to the haploid sexual and diploid sexual case, in the style of Metz and Gyllenberg [Metz, J.A.J., Gyllenberg, M., 2001. How should we define fitness in structured metapopulation models? Including an application to the calculation of ES dispersal strategies. Proc. R. Soc. Lond. B 268, 499-508], that can cope with local population fluctuations due to environmental and demographic stochasticity. With the use of this fitness proxy we find that in dispersal evolution the studied clonal model is equivalent with the haploid sexual model, and that there are indeed many differences between clonal and diploid ESS dispersal rates. In a homogenous landscape the discrepancy is but minor (less than 2%), but the situation is different in a heterogeneous landscape: Not only is the quantitative discrepancy between the two types of ESSs appreciable (around 10%-20%), but more importantly, at the same parameter values, evolutionarily stability properties may differ. It is possible, that the singular strategy is evolutionarily stable in the clonal case but not in the diploid case, and vice versa. PMID- 18384829 TI - The effect of haem biosynthesis inhibitors and inducers on intestinal iron absorption and liver haem biosynthetic enzyme activities. AB - The relation between haem biosynthesis and intestinal iron absorption is not well understood, we therefore investigated the effect of compounds that alter haem metabolism on duodenal iron absorption. CD1 mice were treated with either an inhibitor (succinyl acetone (SA)) or stimulator (2-allyl-2-isopropylacetamide (AIA)) of haem biosynthesis. 5-Aminolaevulinic acid (ALA) dehydratase and urinary ALA and porphobilinogen (PBG) levels, were determined. Intestinal iron absorption was assayed with in vivo and in vitro techniques. Liver hepcidin (Hamp1) and duodenal iron transporter mRNA levels were measured using RT-PCR. AIA caused increased hepatic ALA synthase (1.6-fold) and ALA dehydratase (1.4-fold, both p<0.005) activities and increased urinary ALA and PBG excretion (2.1- and 1.4 fold, p<0.005, p<0.05, respectively). In vivo intestinal iron absorption was reduced to 49% of control (p<0.005). Mice treated with SA showed decreased urinary ALA and PBG levels (75 and 55% control, both p<0.005) and reductions in both ALA synthase and ALA dehydratase activities (77 and 56% control, p<0.05, p<0.005, respectively) in the liver. Liver and duodenal haem and cytochrome oxidase levels were not significantly decreased. Iron absorption was enhanced (1.26-fold, p<0.05) and hepatic Hamp1 mRNA was reduced (53% of control, p<0.05). In vitro duodenal iron uptake after mice were injected with SA also demonstrated an increase in Fe(III) reduction and uptake (1.27- and 1.41-fold, p<0.01 respectively). Simultaneous injections of SA and ALA blocked the enhancing effect on iron absorption seen with SA alone. We conclude that alterations in haem biosynthesis can influence iron absorption and in particular, the intermediate ALA seems to be an inhibitor of iron absorption. PMID- 18384828 TI - Lung response to coarse PM: bioassay in mice. AB - Particulate matter (PM) elicits inflammatory and toxic responses in the lung specific to its constituents, which can vary by region, time, and particle size. To identify the mechanism of toxicity in PM collected in a rural area in the San Joaquin Valley of Central California, we studied coarse particles of 2.5-10 mum diameter (PM(2.5)-PM(10)). Potential pro-inflammatory and toxic effects of PM(2.5)-PM(10) in the lung were investigated using intratracheally instilled mice. We determined total and differential cell profiles and inflammatory chemokines in lung lavage fluid, and biomarkers of toxicity resulting from coarse PM exposure. Responses of the mice were readily observed with total doses of 25 50 mug of PM per mouse. Changes in pro-inflammatory cellular profiles and chemokines showed both dose and time responses; peak responses were observed 24 h after PM instillation, with recovery as early as 48 h. Furthermore, macrophage inflammatory protein (MIP-2) profiles following PM exposures were correlated to levels of measured macrophages and neutrophils recovered from lung lavage fluid of PM-treated animals. Our data suggest that pro-inflammatory effects observed from coarse PM collected during the summer months from California's hot and dry Central Valley are driven largely by the insoluble components of the PM mixture, and are not caused by endotoxin. PMID- 18384830 TI - Multivalent binding of ricin to bovine serum albumin-based neoglycoconjugates. AB - Ricin, a ribosome-inactivating protein from the plant Ricinus communis, is a heterodimeric protein. The A chain is a N-glycosidase and the B chain (RTB) is a lectin with two carbohydrate binding sites. RTB has been shown to bind asialofetuin with much greater affinity than it does galactose, supporting the hypothesis that it may exhibit multivalency. To test this, neoglycoconjugates were prepared and tested for their ability to function as ligands for ricin binding. Because the two carbohydrate binding sites on RTB are approximately 70A apart, bovine serum albumin (BSA) was used as the carbohydrate carrier. It was derivatized with either the oligosaccharide portion of asialo-GM1 or with lactose. These sugars were used because ricin was found to adhere more effectively to asialo-GM1 and LacCer immobilized on plastic than to the other glycosphingolipids tested. Results of binding studies done using surface plasmon resonance indicated that the RTB subunit of ricin exhibited a multivalent effect when it bound to the neoglycoconjugates. PMID- 18384831 TI - Exosome-like vesicles in Gloydius blomhoffii blomhoffii venom. AB - Exosomes are small membrane vesicles (30-100 nm) with an endosome-derived limiting membrane that are secreted by a diverse range of cell types. We provide here the first evidence for the presence of exosome-like vesicles in snake venom. We isolated vesicles from fresh venom from Gloydius blomhoffii blomhoffii by gel filtration. We found that the vesicles showed a typical exosome-like size and morphology as analyzed by electron microscopy. We observed that the vesicles contained dipeptidyl peptidase IV, aminopeptidase A, ecto-5'-nucleotidase and actin. Vesicle preparations truncated bioactive peptides such as angiotensin II, substance P, cholecystokinin-octapeptide, glucose-dependent insulinotropic polypeptide and glucagon-like peptide-1. The role of these vesicles is still unknown, but they may affect blood pressure and glucose homeostasis following envenomation. PMID- 18384832 TI - Effect of ultrasound therapy on the repair of gastrocnemius muscle injury in rats. AB - The aim of this study was to evaluate the effect of the pulsed ultrasound therapy (PUT) in stimulating myoregeneration and collagen deposition in an experimental model of lacerative gastrocnemius muscle lesion in 30 Wistar rats. Fifteen rats were treated (TG) daily with 1 MHz pulsed ultrasound (50%) at 0.57 W/cm(2) for 5 min, and 15 were control animals (CG). Muscle samples were analyzed on postoperative days 4, 7 and 14 through H&E, Picrosirius-polarization and immunohistochemistry for desmin. The lesions presented similar inflammatory responses in both treated and control groups. The areal fraction of fibrillar collagen was larger in the TG at 4 days post-operatively (17.53+/-6.2% vs 6.79+/ 1.3%, p=0.0491), 7 days (31.07+/-7.45% vs 12.57+/-3.6%, p=0.0021) and 14 days (30.39+/-7.3% vs 19.13+/-3.51%, p=0.0118); the areal fraction of myoblasts and myotubes was larger in the TG at 14 days after surgery (41.66+/-2.97% vs 34.83+/ 3.08%, p=0.025). Our data suggest that the PUT increases the differentiation of muscular lineage cells, what would favor tissue regeneration. On the other hand, it is also suggested that there is a larger deposition of collagenous fibers, what could mean worse functional performance. However, the percentage of fibers seems to have stabilized at day 7 in TG and kept increasing in CG. Furthermore, the collagen supramolecular organization achieved by the TG is also significant according to the Sirius red staining results. PMID- 18384834 TI - The assessment of visual acuity in children with periventricular damage: a comparison of behavioural and electrophysiological techniques. AB - It has been controversial whether electrophysiology offers better precision than behavioural techniques in measuring visual acuity in children with brain damage. We investigated the concordance between sweep VEPs and Acuity Cards (AC) in 29 children with periventricular leukomalacia (PVL), the most common type of brain damage in preterm infants. An overall good correlation was shown but with relatively better behavioural acuity values. VEP/AC ratio was significantly correlated to corpus callosum posterior thinning. We propose that this result reflects the efficacy of the compensatory mechanisms following early brain damage which may differentially affect the two methods. PMID- 18384833 TI - Endogenous attention selection during binocular rivalry at early stages of visual processing. AB - Directing attention to one of two superimposing surfaces composed of dot fields rotating in opposing directions facilitates processing of brief translations of the attended surface [Valdes-Sosa, M., Bobes, M. A., Rodriguez, V., & Pinilla, T. (1998). Switching attention without shifting the spotlight object-based attentional modulation of brain potentials. Journal of Cognition and Neuroscience, 10(1), 137-151]. Here we used ERP recordings to investigate the mechanisms of endogenous attentional selection of such competing dot surfaces under conditions of dichoptic viewing (one surface to each eye) and monocular viewing (both surfaces to one eye). Under dichoptic conditions, which induced binocular rivalry, translations of the attended surface presented to one eye elicited enhanced visual P1 and N1 ERP components relative to translations of the unattended surface presented to the other eye. In comparison, during monocular viewing the attended surface translations elicited a significantly larger N1 component in the absence of any P1 modulation. These results indicate that processing of the attended surface is biased at an earlier level in extrastriate visual cortex under conditions of inter-ocular versus intra-ocular competition. PMID- 18384835 TI - Chemical and microbial decontamination of pool water using activated potassium peroxymonosulfate. AB - Potassium peroxymonosulfate activation leads to the formation of highly reactive species, mainly the sulfate radicals. Activated potassium peroxymonosulfate (from now on peroxymonosulfate) was tested against specific pollutants such as ammonium ion, creatinine, chlorinated creatinine products, arginine and Escherichia coli (E. coli), all constituents or derivatives of human discharges. The objective was to assess whether activated peroxymonosulfate can be a viable treatment reagent in recreational water applications. It was found that organic molecules such as creatinine, chlorinated creatinine products and arginine could be effectively treated with activated peroxymonosulfate. Ammonium ion was oxidized only by chlorine species and only in de-ionized water. Chlorine species were formed from the reaction of sulfate radicals with chloride ions. In pool water, the reaction of sulfate radicals with chloride ions and the subsequent ammonium ion oxidation were scavenged by the presence of bicarbonate ions. The Co/Peroxymonosulfate system was also shown to be an effective disinfection reagent, since 99.99% (4 log) kill of E. coli was achieved in 60 min of treatment. At the concentrations tested here, however, it is still not efficacious enough to qualify as an EPA registered sanitizer for swimming pools (requires 6-log kill of E. coli, ATCC 11229, and Enterococcus faecium, ATCC 6569, in 30s). PMID- 18384836 TI - Photochemical formation of hydroxyl radicals catalyzed by montmorillonite. AB - In this work, the photooxidation of benzene and the formation of phenol in aqueous suspensions of the iron-rich montmorillonite under irradiation of a 250W metal halide lamp (lambda> or = 365nm) were investigated. We confirmed that hydroxyl radicals (()OH) were produced by illuminating montmorillonite and was responsible for the photooxidation of benzene in aqueous suspensions of montmorillonite. Low pH value facilitated the formation of hydroxyl radicals in the pH range of 2.0-10.0. The ()OH concentration increased with increasing the concentration of montmorillonite in aqueous solutions in the range of 0-20.0gl( 1). Higher concentration like 25.0gl(-1) montmorillonite inhibited the ()OH production. Iron, predominantly free iron in the clays, is believed to be one of the most important factors determining ()OH formation. Structural irons in montmorillonite have contributions to ()OH formation, especially in the presence of carboxylate ions. The formation of ()OH from montmorillonite under irradiation of near UV and visible light indicates that clays might play important role not only in transfer through adsorption but also in transformation through oxidation of organic compounds on the surface of clay particles in air, water, soil or even top sediments. PMID- 18384837 TI - A comparison of DEMETRA individual QSARs with an index for evaluation of uncertainty. AB - Quantitative structure-activity relationships (QSARs) urgently need to be applied in regulatory programs. Many QSAR models can predict the effect of a wide range of substances to different endpoints, particularly in the case of ecotoxicity, but it is difficult to choose the most appropriate model on the basis of the requirements of the application. During the EC-funded project DEMETRA (www.demetra-tox.net) a huge number of QSAR models have been developed for the prediction of different ecotoxicological endpoints. DEMETRA individual models on rainbow trout LC50 after 96 h, water flea LC50 after 48 h and honey bee LD50 after 48 h have been used as a QSAR database to test the advantages of a new index for evaluating model uncertainty. This index takes into consideration the number of outliers (weighted on the total number of compounds) and their root mean square error. Application on the DEMETRA QSAR database indicated that the index can identify the models with the best performance with regard to outliers, and can be used, together with other classical statistical measures (e.g., the squared correlation coefficient), to support the evaluation of QSAR models. PMID- 18384838 TI - The speciation, stability, solubility and biodegradation of organic co contaminant radionuclide complexes: a review. AB - The potential migration of radionuclides is of concern at contaminated land sites and, in the long term, waste repositories. Pathways of migration need to be characterised on a predictive level so that management decisions can be made with confidence. A pathway that is relatively poorly understood at present is radionuclide solubilisation due to complexation by organic complexing agents that are present in mixed radioactive wastes, and at radioactively contaminated land sites. Interactions of the complexing agents with radionuclides and the host environment, and the response to changes in the physicochemical conditions make their role far from simple to elucidate. In addition, chemical and biodegradation of the organic materials may be important. In this paper, key co-contaminant organics are reviewed with emphasis on their environmental fate and impact on radionuclide migration. PMID- 18384839 TI - Organochlorine contaminants in endangered Steller sea lion pups (Eumetopias jubatus) from western Alaska and the Russian Far East. AB - Investigations into the cause of the Steller sea lion population decline have focused on numerous factors, including exposure to toxic contaminants such as organochlorines (OCs). OCs, such as polychlorinated biphenyls (PCBs), 1,1,1 trichloro-2,2-bis(p-chlorophenyl)ethane, or dichlorodiphenyltrichloroethane (DDT), have been associated with various biological effects in marine mammals. We measured these compounds in whole blood of free-ranging Steller sea lion pups in order to determine the extent and magnitude of contamination across their geographical range. Of 212 pups analyzed for OCs, 76 pups (36 females and 40 males) were from western Alaska and the other 136 (63 females and 73 males) were from the Russian Far East. Concentrations of summation SigmaPCBs in the whole blood of pups from western Alaska ranged from 0.21 to 13 ng/g wet weight with a mean of 2.1+/-0.27 ng/g wet weight. In the Russian animals, summation SigmaPCB concentrations in the whole blood of pups ranged from 0.33 to 36 ng/g wet weight with a mean of 4.3+/-0.44 ng/g wet weight. summation SigmaDDT concentrations in the whole blood of pups from western Alaska ranged from 0.18 to 11 ng/g wet weight with a mean of 1.6+/-0.23 ng/g wet weight. In Russia, summation SigmaDDT in the whole blood of pups ranged from undetectable to 26 ng/g wet weight with a mean of 3.3+/-0.36 ng/g wet weight. Average OC concentrations were significantly higher in the blood of Russian animals compared to western Alaska (for PCBs and DDTs, p<0.001) and in both areas females had higher concentrations than males. Male pups from western Alaska had significantly lower levels of summation SigmaPCBs and summation SigmaDDT when compared to male pups from Russia (for PCBs and DDTs p<0.001). Female pups from western Alaska were significantly lower in summation SigmaPCBs than Russian female pups (for PCBs p=0.009) as were female pups for summation SigmaDDT levels between areas (for DDTs p=0.026). OC contaminants data indicate that Steller sea lion pups have measurable concentrations of these synthetic chemicals. While any physiological effect and the specific role these chemicals may have in either the decline or the failure of the endangered Steller sea lion population to recover needs to be further investigated, this study indicates specific areas and animals that may be most at risk. PMID- 18384840 TI - Relationship between maternal and cord blood hemostatic disturbances in preeclamptic pregnancies. AB - Endothelial cell activation or damage is believed to play a key role in preeclampsia (PE) and may underlie the hemostatic changes observed in this syndrome. The aim of this study was to evaluate a relationship between maternal and cord blood hemostatic disturbances in preeclamptic pregnancies. We measured the plasma levels of tissue plasminogen activator (tPA) antigen and of plasminogen activator inhibitor type 1 (PAI-1) antigen, both markers of hemostatic and endothelial function, and fibrin fragment D-dimer. Maternal blood from uncomplicated (n=42) and PEc pregnancies (n=44) were collected before delivery, and umbilical cord blood (UCB) immediately after delivery. In preeclamptic cases, UCB presented significantly higher tPA values and significantly lower PAI-1/tPA ratio. Preeclamptic women also presented significantly higher tPA, as well as PAI-1 values, when compared with normal pregnant women; no significant difference was found for D-dimer. In preeclamptic women, proteinuria (a marker of PE severity) correlated positively and significantly with tPA and PAI-1 antigen levels. An inverse relationship between maternal tPA antigen levels and fetal birth weigh in PE was also observed. Our data show that the hemostatic maternal disturbances observed in preeclamptic women have similarities with the UCB circulation, and that endothelial dysfunction is the most plausible underlying cause. Moreover, maternal hemostatic disturbances seem to be associated with the severity of PE. Further studies are needed to strength the values of tPA and PAI-1 as markers of severity in PE. PMID- 18384841 TI - The in vitro anticoagulant effects of danaparoid, fondaparinux, and lepirudin in children compared to adults. AB - INTRODUCTION: Major physiological differences in the coagulation system of children compared to that of adults are well documented. We have previously investigated the age-related differences in response to Unfractionated Heparin (UFH). However, the impact of developmental haemostasis on more recent anticoagulant drugs is unknown. A number of these drugs are approved for use in specific indications in adults and none are approved for use in children. This study aimed to determine whether age-related differences in effect and impact on monitoring tests exist in vitro for danaparoid, fondaparinux and lepirudin. MATERIALS AND METHODS: Plasma samples were obtained from healthy children and pooled into age-specific pools, in order to obtain sufficient quantity of plasma required for the analysis of the three drugs. Each age-specific pool was spiked with different concentrations of danaparoid, fondaparinux and lepirudin and response was measured using standard techniques. All experiments were repeated using three separate plasma pools. The effect of each drug in children's plasma was compared to the effect in the respective adult plasma pool. RESULTS: Age related differences in effect on thrombin potential and monitoring tests were observed only with the drug lepirudin. Specifically, APTT for children up to 5 years of age was increased compared to adults; all children had lower ECT results compared to adults; children up to 10 years of age had increased inhibition of ETP as compared to adults. CONCLUSIONS: This study confirms age-related differences in response to anticoagulants with predominant anti-IIa effect and highlights the need for further research into this area. PMID- 18384842 TI - Methylenetetrahydrofolate reductase gene, homocysteine and coronary artery disease: the A1298C polymorphism does matter. Inferences from a case study (Madeira, Portugal). AB - Elevated levels of plasma homocysteine, an independent risk factor and a strong predictor of mortality in patients with coronary artery disease (CAD), can result from nutritional deficiencies or genetic errors, including methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C polymorphisms. The contribution of these polymorphisms in the development of CAD remains controversial. We analysed the impact of MTHFR C677T and A1298C on fasting homocysteine and CAD in 298 CAD patients proved by angiography and 510 control subjects from the Island of Madeira (Portugal). After adjustment for other risk factors, plasma homocysteine remained independently correlated with CAD. Serum homocysteine was significantly higher in individuals with 677TT and 1298AA genotypes. There was no difference in the distribution of MTHFR677 genotypes between cases and controls but a significant increase in 1298AA prevalence was found in CAD patients. In spite of the clear effect of C677T mutation on elevated homocysteine levels we only found an association between 1298AA genotype and CAD in this population. The simultaneous presence of 677CT and 1298AA genotypes provides a significant risk of developing the disease, while the 1298AC genotype, combined with 677CC, shows a significant trend towards a decrease in CAD occurrence. The data shows an independent association between elevated levels of homocysteine and CAD. Both MTHFR polymorphisms are associated with increased fasting homocysteine (677TT and 1298AA genotypes), but only the 1298AA variant shows an increased prevalence in CAD group. Odds ratio seem to indicate that individuals with the MTHFR 1298AA genotype and the 677CT/1298AA compound genotype had a 1.6-fold increased risk for developing CAD suggesting a possible association of MTHFR polymorphisms with the risk of CAD in Madeira population. PMID- 18384843 TI - Epidermal nerve fiber quantification in the assessment of diabetic neuropathy. AB - Assessment of cutaneous innervation in skin biopsies is emerging as a valuable means of both diagnosing and staging diabetic neuropathy. Immunolabeling, using antibodies to neuronal proteins such as protein gene product 9.5, allows for the visualization and quantification of intraepidermal nerve fibers. Multiple studies have shown reductions in intraepidermal nerve fiber density in skin biopsies from patients with both type 1 and type 2 diabetes. More recent studies have focused on correlating these changes with other measures of diabetic neuropathy. A loss of epidermal innervation similar to that observed in diabetic patients has been observed in rodent models of both type 1 and type 2 diabetes and several therapeutics have been reported to prevent reductions in intraepidermal nerve fiber density in these models. This review discusses the current literature describing diabetes-induced changes in cutaneous innervation in both human and animal models of diabetic neuropathy. PMID- 18384844 TI - Effects of maternal undernutrition on renal angiotensin II and chymase in hypertensive offspring. AB - Intrauterine growth restriction (IUGR) can program the future development of hypertension in adulthood. The renin-angiotensin system has been reported to play a role in IUGR-induced hypertension. The aims of this study were to investigate the effects of IUGR on renal angiotensin-converting enzyme (ACE), angiotensin II (Ang II) and chymase in IUGR-induced hypertension. Timed pregnant Sprague-Dawley rats received 50% rations of control food intakes from days 15 to 21 of gestation. Control rats received regular food throughout the pregnancies. Arterial blood pressure and glomerular number were measured and immunohistochemical studies were performed on kidney tissues in adult male offspring at 16 weeks of age. IUGR rats exhibited significantly lower body and kidney weights and reduced number of glomeruli when compared with control rats. IUGR rats had significantly higher systolic blood pressure than control rats. Immunoreactivity of ACE was comparable between control and IUGR rats whereas immunoreactivities of chymase and Ang II were significantly higher in IUGR rats than in control rats. In conclusion, immunohistochemical studies document up regulation of ACE-independent Ang II and chymase in IUGR kidney and indicate that overactivity of chymase may result in increased intrarenal Ang II production, which could contribute to the development of hypertension in intrauterine undernourished rats. PMID- 18384845 TI - Energetic consequences of mechanical loads. AB - In this brief review, we have focussed largely on the well-established, but essentially phenomenological, linear relationship between the energy expenditure of the heart (commonly assessed as the oxygen consumed per beat, oxygen consumption (VO2)) and the pressure-volume-area (PVA, the sum of pressure-volume work and a specified 'potential energy' term). We raise concerns regarding the propriety of ignoring work done during 'passive' ventricular enlargement during diastole as well as the work done against series elasticity during systole. We question the common assumption that the rate of basal metabolism is independent of ventricular volume, given the equally well-established Feng- or stretch effect. Admittedly, each of these issues is more of conceptual than of quantitative import. We point out that the linearity of the enthalpy-PVA relation is now so well established that observed deviations from linearity are often ignored. Given that a one-dimensional equivalent of the linear VO2-PVA relation exists in papillary muscles, it seems clear that the phenomenon arises at the cellular level, rather than being a property of the intact heart. This leads us to discussion of the classes of crossbridge models that can be applied to the study of cardiac energetics. An admittedly superficial examination of the historical role played by Hooke's Law in theories of muscle contraction foreshadows deeper consideration of the thermodynamic constraints that must, in our opinion, guide the development of any mathematical model. We conclude that a satisfying understanding of the origin of the enthalpy-PVA relation awaits the development of such a model. PMID- 18384847 TI - Effect of bundle branch block on cardiac output: a whole heart simulation study. AB - The heart is an electrically controlled fluid pump which operates by mechanical contraction. Whole heart modelling is a computationally daunting task which must incorporate several subsystems: mechanical, electrical, and fluidic. Numerous feedback mechanisms on many levels, and operating at different scales, exist to finely control behaviour. Understanding these interactions is necessary to understand heart operation, as well as pathologies and therapies. A review of the components in such a model is given. The authors then present a framework for their electro-mechano-fluidic whole heart model based on cable methods. The model incorporates atria and ventricles, and has functioning valves with papillary muscles. The effect of altered propagation due to left and right bundle branch block on cardiac output is examined using the cable-based model. Results are compared to clinically observed phenomena. Good agreement was obtained, but tighter coupling of mechanical and electrical events is needed to fully account for behaviour. Cable-based models offer an alternative to continuum models. PMID- 18384846 TI - Cell cultures as models of cardiac mechanoelectric feedback. AB - Although stretch-activated currents have been extensively studied in isolated cells and intact heart in the context of mechanoelectric feedback (MEF) in the heart, quantitative data regarding other mechanical parameters such as pressure, shear, bending, etc, are still lacking at the multicellular level. Cultured cardiac cell monolayers have been used increasingly in the past decade as an in vitro model for the studies of fundamental mechanisms that underlie normal and pathological electrophysiology at the tissue level. Optical mapping makes possible multisite recording and analysis of action potentials and wavefront propagation, suitable for monitoring the electrophysiological activity of the cardiac cell monolayer under a wide variety of controlled mechanical conditions. In this paper, we review methodologies that have been developed or could be used to mechanically perturb cell monolayers, and present some new results on the acute effects of pressure, shear stress and anisotropic strain on cultured neonatal rat ventricular myocyte (NRVM) monolayers. PMID- 18384848 TI - Expression of CD24 in clear cell renal cell carcinoma and its prognostic significance. AB - OBJECTIVES: To evaluate the expression patterns of CD24 in clear cell renal cell carcinoma (CCRCC) and to investigate its prognostic significance. CD24 is a cell adhesion molecule that has been thought to play an important role in tumor progression and metastasis of various cancers. CD24 expression has previously been reported in RCC, although little is known about its prognostic significance. METHODS: We immunohistochemically analyzed CD24 expression using tissue microarray in 328 cases of CCRCC. The percentage of positively stained tumor cells was evaluated and classified into four categories: 0, 0%; 1+, 1% to 10%; 2+, 11% to 50%; and 3+, more than 50%. For statistical analysis, the cases were subdivided into a CD24-low-expression group (0 and 1+) and CD24-high-expression group (2+ and 3+). RESULTS: High CD24 expression was significantly associated with a high nuclear grade (P = 0.005) and large tumor size (P = 0.04). The survival analysis revealed no significant association between CD24 expression and disease-specific survival (P = 0.141). However, the CD24-high-expression group had a significantly shortened progression-free survival (P = 0.005). Multivariate analysis also revealed that high CD24 expression was an independent covariate for poor progression-free survival (P = 0.043). CONCLUSIONS: We investigated CD24 expression in CCRCC and found that high CD24 expression was associated with high nuclear grade, large tumor size, and shortened progression-free survival. Although the biologic function of CD24 in CCRCC remains unknown, the expression of CD24 can provide new prognostic information about disease progression. PMID- 18384849 TI - Does "normal" aging imply urinary, bowel, and erectile dysfunction? A general population survey. AB - OBJECTIVES: We assessed if urinary, bowel, and sexual dysfunction and associated bother were part of the "normal" aging process in the general male Dutch population. METHODS: Randomly selected participants of a screening trial were mailed a questionnaire on dysfunction and bother in the urinary, bowel, and sexual domains. A Dutch version of the Expanded Prostate Cancer Index Composite (EPIC) was used. RESULTS: Three thousand eight hundred ten (3810) men responded (81%), mean age 67 years, range 58 to 78. The prevalence of urinary dysfunction was low, and although the difference between younger versus older men was significant (P <0.001), it did not exceed the minimal important difference. Bowel dysfunction and bother were not related to age. Erectile dysfunction was reported by 19%, ranging from 12% in the youngest to 26% in the oldest group (P <0.001). The overall use of erectile aids was negatively associated with the satisfaction with sex life and positively with the importance attached to it (P <0.001), but not with age or sexual activity. CONCLUSIONS: Urinary and bowel dysfunction were not part of the "normal" aging process. Erectile dysfunction was significantly more prevalent in older men. In men treated for localized prostate cancer, decreasing urinary or bowel function is thus not attributable to age, but may well be related to prior treatment. Decreasing erectile function, however, may be attributable to other causes as well. These data provide a benchmark for urologic functioning in men after treatment relative to age-related patterns, and will enable better interpretation of treatment outcomes. PMID- 18384850 TI - Differential expression profile of cold (TRPA1) and cool (TRPM8) receptors in human urogenital organs. AB - OBJECTIVES: To examine the expressions of transient receptor potential (TRP) channel A1 and TRPM8 in the human urogenital organs (urinary bladder and prostate) and how these expressions change in association with bladder outlet obstruction (BOO) or benign prostatic hyperplasia (BPH). In addition to TRPM8, a cool receptor, TRPA1 is recently recognized as a cold receptor. TRPA1 is also a candidate for mechanosensor and/or nociceptor. METHODS: Urinary bladder mucosa and muscular layer were taken from 9 controls and 9 patients with BOO. Prostatic specimens were obtained from 5 controls and 6 patients with BPH. Expressions of TRPA1 and TRPM8 messenger RNAs (mRNAs) were quantified by real-time revere transcriptase-polymerase chain reaction. Localization of TRPA1 protein was explored with immunohistochemistry. RESULTS: The expression levels of TRPA1 mRNA in the bladder mucosa, bladder muscular layer, and prostate were in the ratio of 639:1:16. TRPA1 mRNA in the bladder mucosa with BOO was significantly upregulated to 2.32 times of control. TRPA1 protein was localized in the epithelial cells of both urinary bladder and prostate gland. The expression of TRPM8 mRNA in the prostate was much higher than that in the bladder mucosa (3024:1), but was not found in the bladder muscle layer. BPH or BOO did not significantly affect the expression of TRPM8. CONCLUSIONS: TRPA1 and TRPM8 were differentially expressed in the human urinary bladder and prostate. TRPA1 in the bladder epithelium might be involved in the bladder sensory transduction and the induction process of overactive bladder by BOO. PMID- 18384851 TI - Simultaneous "cut and suture": a novel technique for laparoscopic partial nephrectomy without vascular clamping. AB - Loss of renal units due to warm ischemia is of great concern in laparoscopic partial nephrectomy (LPN). We describe a technique for performing LPN without vascular clamping with optimal results and minimal blood loss, suitable for small and superficial renal masses. PMID- 18384852 TI - Granular cell tumor of scrotum: a rare tumor of the male external genitalia. AB - We report a rare case of granular cell tumor in the scrotum. Granular cell tumors are soft-tissue neoplasms originating from Schwann cells that rarely affect male external genitalia. They are essentially benign; therefore, the treatment is complete excision of the lesion. Although never previously reported in the male external genitalia, malignant variants exist in 2% of cases. Because the clinical presentation is not specific, the diagnosis of malignant granular cell tumors can be made only by the pathologist. To our knowledge, only 5 other cases in the scrotum and 19 cases described in the penis have been reported. PMID- 18384853 TI - Return to continence after radical retropubic prostatectomy: a randomized trial of verbal and written instructions versus therapist-directed pelvic floor muscle therapy. AB - OBJECTIVES: To test the effectiveness of weekly postoperative pelvic floor muscle training (PFMT) versus supportive telephone contact by a urology nurse for men at 4 weeks after radical prostatectomy. METHODS: This was a randomized controlled trial in three Canadian centers. At 4 weeks after surgery, standardized verbal and written instruction about PFMT was provided to all subjects. Randomization occurred after initial instruction. Continence was defined as 8 g or less of urine loss on a 24-hour pad test. Primary outcome was grams of urine loss on pad test; secondary outcomes were International Prostate Symptom Score (IPSS), Incontinence Impact Questionnaire (IIQ-7) score, cost, and perception of urine loss as a problem. Data were obtained at baseline (preoperatively) and at weeks 4, 8, 12, 16, and 28 and 1 year after surgery. RESULTS: A total of 216 men were enrolled; 11 were dry or withdrew at 4 weeks. Ninety-nine were randomized to the control group and 106 to the treatment group. There were no group differences at baseline for prostate-specific antigen level (mean [standard deviation] 8.4 [10.4] ng/mL; 7.6 [4.6] ng/mL), Gleason score (6.3 [0.86]), IPSS, IIQ-7 score, pad test, or voiding diary. At 8 weeks 23% of the control group and 20% of the treatment group were continent; at 12 weeks, 28% and 32%; 16 weeks, 40% and 44%; 28 weeks, 50% and 47%; and at 52 weeks, 64% and 60%, respectively. There were no significant differences between groups at any time point for the outcome variables. CONCLUSIONS: Verbal instruction and written information with telephone support seemed to be as effective as intensive PFMT. Less-intense therapy may be more cost-effective. PMID- 18384854 TI - Malfunction of da Vinci robotic system--disassembled surgeon's console hand piece: case report and review of the literature. AB - Recently, increasing numbers of robotic-assisted laparoscopic radical prostatectomy have been performed at many centers. Although uncommon, malfunction of the da Vinci Surgical system represents a new and unique problem in urologic surgery. In this study, we report a rare case of a disassembled surgeon's console hand piece because of a loose screw during robotic-assisted laparoscopic radical prostatectomy. PMID- 18384855 TI - A pure triphasic testicular wilms tumor of primordial germ cell origin. PMID- 18384856 TI - Utility of Bcl-2, P53, Ki-67, and caveolin-1 immunostaining in the prediction of biochemical failure after radical prostatectomy in a Japanese population. AB - OBJECTIVES: To identify predictive markers for biochemical failure after radical prostatectomy in patients with clinically confined prostate cancer. METHODS: Immunohistochemistry of bcl-2, p53, Ki-67, and caveolin-1 was performed in samples of paraffin-embedded prostate cancer from 119 Japanese patients. The clinicopathologic significance of staining with these markers was analyzed in relation to biochemical failure (prostate-specific antigen [PSA] >0.2 ng/mL). RESULTS: Univariate analysis showed the pretreatment PSA level (P = 0.03), postoperative Gleason score (P = 0.04), pathologic stage (P <0.001), seminal vesicle invasion (P <0.001), p53 staining (P <0.001), Ki-67 staining (P = 0.04), and caveolin-1 staining (P <0.0001) to be associated with biochemical failure. Multivariate Cox proportional hazards modeling showed that pretreatment PSA in group A (clinicopathologic parameters), caveolin-1 staining in group B, biomarkers, and the combination (group C) were independently associated with prediction of biochemical failure. The accuracy rate of each group was 76.2% (group A), 75.1% (group B), and 83.1% (group C), respectively. CONCLUSIONS: The combination of clinicopathologic parameters and biomarkers (group C) showed the highest accuracy rate. Caveolin-1 staining is an independent predictor of biochemical failure after radical prostatectomy. PMID- 18384857 TI - Significant discrepancies between diagnostic and pathologic Gleason sums in prostate cancer: the predictive role of age and prostate-specific antigen. AB - OBJECTIVES: To assess the discrepancies between diagnostic and pathologic Gleason sums and the predictive role of age and prostate-specific antigen (PSA) level on Gleason sum discrepancies. METHODS: A total of 2963 patients receiving radical prostatectomy at Duke University from 1988 to 2006 were divided into two groups according to year of diagnosis: 1988 to 1999 and 2000 to 2006. The Gleason sum discrepancies were evaluated in the above groups. The predictive roles of diagnostic age (less than 50, 50 to 60, 60.1 to 70, and greater than 70 years), PSA level (less than 10, 10 to 20, and greater than 20 ng/mL), race, body mass index, and prostate weight on the discrepancies were analyzed. RESULTS: Overall, 55.8% of diagnostic Gleason sums differed from those on final surgical pathology (58.6% in the 1988 to 1999 and 49.3% in the 2000 to 2006 groups). Diagnostic Gleason sums were undergraded in 41.2% of cases and overgraded in 12.8% of cases. Men older than 60 years were more likely to have their diagnostic Gleason sums undergraded than men younger than 50 (odds ratio in age groups less than 50, 50 to 60, 60.1 to 70, and greater than 70 years: 1.00, 2.30, 4.03, and 3.96, respectively). Biopsy Gleason sums in men with a high PSA level were more likely to be undergraded compared with the PSA group less than 10 ng/mL (odds ratio in PSA groups less than 10, 10 to 20, and greater than 20 ng/mL: 1.00, 2.11, and 3.64, respectively). CONCLUSIONS: Significant discrepancies between diagnostic and pathologic Gleason sums remain in recent years. The rate of diagnostic Gleason sum undergrading was 3.2-fold that of overgrading. Advanced age and high PSA level were predictive of diagnostic Gleason sum undergrading, and caution should be exercised when recommending active surveillance in older men. PMID- 18384858 TI - Robotic-assisted laparoscopic ureteral reimplantation with psoas hitch: a multi institutional, multinational evaluation. AB - OBJECTIVES: To report the collective experience of three multinational institutions with the use of robotics to evaluate and treat complex distal ureteral obstruction. METHODS: A total of 12 patients from The Ohio State University, Columbus, Ohio; Onze-Lieve-Vrouw Ziekenhuis, Aalst, Belgium; and Hospital Sultanah Aminah, Kuala Lumpur, Malaysia underwent robotic-assisted laparoscopic ureteral reimplantation between August 2004 and July 2006. The indications for ureteral reimplantation included ureteral stricture (n = 10) and ureterovaginal fistula (n = 2). Nine patients had pathology on the left side and 4 patients had right-sided disease. Surgery was performed by three experienced laparoscopic robotic surgeons with the daVinci Surgical System. RESULTS: The mean patient age (range) was 41.3 years (19 to 67 years). The mean operative time was 208 minutes (80 to 360 minutes). The mean robot time was 173 minutes (75 to 300 minutes). The mean estimated blood loss was 48 mL (45 to 100 minutes). The mean length of hospitalization was 4.3 days (2 to 8 days). All the procedures were completed successfully robotically without open conversion. There were no intraoperative or postoperative complications. Postoperative intravenous urography and Mercapto Acetyl TriGlycine 3 showed normal findings in 10 patients and a mild residual hydronephrosis in 2 patients. After a mean follow-up of 15.5 months, all patients were asymptomatic of their initial disease state. CONCLUSIONS: This multi-institutional, multinational experience illustrates that ureteral reimplantation with psoas hitch can be performed safely and effectively to treat lower tract ureteral obstruction. PMID- 18384859 TI - Bladder cancer in Native Americans and Alaskan Natives. AB - OBJECTIVES: The prevalence of bladder cancer among Native Americans/Alaskan Natives (NAs/ANs) is generally considered to be low. However, the relative morbidity and mortality seem high. We have undertaken this survey of the published data to explore and document this observation. METHODS: An extensive survey of the published data was undertaken. The Surveillance, Epidemiology and End Results data in particular were assessed. RESULTS: The incidence of bladder cancer among the NA/AN peoples is inexplicably low. This low incidence is more remarkable, given the widespread use of tobacco products and the disproportionately high incidence of kidney cancer in this same population. Despite this low incidence, NA/AN men and women seem to be at a relatively greater risk of dying of bladder cancer, once it has been diagnosed. CONCLUSIONS: More and larger studies are needed to clarify and update the incidence, cell type, and clinical outcomes of bladder cancer among NA/AN men and women. Social, economic, and political barriers to treatment also need to be addressed. Effective steps need to be undertaken to ensure more timely diagnosis and treatment of this disease. PMID- 18384860 TI - BCGosis: case report of intravesicular bacille Calmette-Guerin causing upper tract granulomas simulating a renal mass without evidence of ipsilateral vesicoureteral reflux. AB - Very rarely, dissemination of intravesical bacille Calmette-Guerin for superficial bladder cancer treatment can lead to a complication in the upper urinary tract. Two mechanisms for dissemination have been proposed: vesicoureteral reflux and hematogenous spread. We report a case of "BCGosis" with the clinical appearance of multifocal renal masses in a patient without ipsilateral reflux. This case highlights the importance of obtaining a renal biopsy in all patients previously treated with bacille Calmette-Guerin presenting with a renal mass. Furthermore, the findings from the present case suggest that vesicoureteral reflux might not be the only mechanism responsible for dissemination of bacille Calmette-Guerin to the upper urinary tract. PMID- 18384861 TI - Ureterovaginal fistula formation after oocyte retrieval. AB - Transvaginal ultrasound-guided oocyte retrieval is generally a safe and well tolerated procedure that is usually associated with few overall complications (less than 5%), including mild vaginal hemorrhage and infection. Injury to the ureter during oocyte retrieval has only been reported in 6 cases to date. We report a case of ureterovaginal fistula that formed approximately 7 days after oocyte retrieval. A percutaneous nephrostomy tube was placed using ultrasound guidance, and the fistula was allowed to close secondarily. Although a rare complication, physicians should be conscious of this type of injury after oocyte retrieval so that prompt diagnosis and treatment can be pursued. PMID- 18384862 TI - Varicocelectomy for infertile couples with advanced paternal age. AB - OBJECTIVES: To evaluate the reproductive outcomes of infertile couples with a clinical varicocele and advanced paternal age. METHODS: We reviewed the clinical records of 581 consecutive, non-azoospermic men presenting with a clinical varicocele and infertility. RESULTS: We identified 115 men aged 40 years and older and 466 men younger than 40 years with a clinical varicocele and infertility. The proportion of men with secondary infertility was significantly higher in the group of men aged 40 years and older compared with the men younger than 40 years (43% [50 of 115] versus 19% [87 of 466], respectively; P <0.001). There were no significant differences in baseline sperm parameters and in spontaneous pregnancy rates after varicocelectomy in couples with advanced paternal age (40 years or older) compared with the younger couples (49% versus 39%, respectively). However, the spontaneous pregnancy rate in couples with advanced paternal age (40 years or older) who underwent varicocelectomy was significantly greater than that of the age-matched control group who did not undergo surgery (49% versus 21%, respectively; P <0.05). CONCLUSIONS: The results of this study suggest that paternal age does not adversely influence pregnancy outcome after varicocelectomy. The data support the practice of varicocelectomy for treatment of clinical varicocele and infertility in older men. PMID- 18384863 TI - Metastatic renal cell carcinoma with partial response to sunitinib complicated by ureteral obstruction from necrotic tumor. AB - The development of molecular therapeutic agents has revolutionized the treatment of metastatic renal cell carcinoma, but the long-term complications of these novel medications are not yet well-defined. We report the case of a 61-year-old woman who presented with metastatic renal cell carcinoma to the liver and contralateral kidney and renal pelvis 6 years after nephrectomy. Treatment with four cycles of sunitinib malate resulted in an initial near-complete radiographic response. Subsequently, the patient presented with flank pain, acute renal failure, and hydronephrosis of her solitary kidney during her fourth cycle of treatment. A temporizing percutaneous nephrostomy tube was placed, and antegrade contrast studies showed a filling defect and complete distal ureteral occlusion. Ureteroscopy revealed ureteral obstruction by a soft-tissue mass. Pathologic examination demonstrated necrotic renal cell carcinoma, presumably due to sloughing of the previously identified renal pelvic metastasis. We discuss the implications of this unusual complication of complete ureteral obstruction from necrotic tumor in a patient with a near-complete response to sunitinib. PMID- 18384864 TI - Human dirofilariasis due to Dirofilaria repens mimicking a scrotal tumor. AB - Dirofilariasis due to Dirofilaria repens is a rare anthropozoonotic disease, with dogs and cats being the definite hosts for this parasite. This worm is transmitted by various species of mosquitoes and usually presents as a subcutaneous nodule. Human cases have been reported from all around the Mediterranean, Eastern Europe, and Central and Southern Asia. In the present case involving a 28-year-old patient of Tunisian origin, infection manifested as a nodule in the left epididymis. In the context of increasing international travel and migration, dirofilariasis is a diagnosis that should be kept in mind when treating patients who have spent some time in endemic areas and present with unclear subcutaneous or scrotal nodules. PMID- 18384865 TI - Enuresis is a common and persistent problem among children and young adults with sickle cell anemia. AB - OBJECTIVES: Enuresis and nocturia are common among children with sickle cell anemia (SCA). The objectives of this study were to describe the prevalence of enuresis and nocturia among children and young adults with SCA and determine the relationship, if any, between these symptoms and SCA-related morbidity. METHODS: A prospective infant cohort of African-American children with SCA was previously established from the Cooperative Study for Sickle Cell Disease. Included in this cohort were children with SCA enrolled before 6 months of age for whom questions about enuresis and nocturia had been completed. RESULTS: A total of 213 participants were included in this analysis. Sixty-nine individuals (33%) experienced enuresis over the course of the study. No children under 6 years of age were asked about enuresis. Thereafter, enuresis was most prevalent between the ages of 6 and 8 years (42%) and continued to be common in young adults ages 18 to 20 years (9%). Seventy-nine percent of individuals reported a history of nocturia. There was no association between enuresis or nocturia and an increased rate of pain or acute chest syndrome (ACS) episodes. CONCLUSIONS: Enuresis and nocturia are common in children with SCA. Among adults with SCA, enuresis and nocturia are more persistent compared with adults in the general population. Enuresis and nocturia are not associated with an increased rate of pain or ACS. PMID- 18384866 TI - Assessment of outcomes and morbidity following diaphragmatic peritonectomy for women with ovarian carcinoma. AB - OBJECTIVE: To describe the technique of diaphragmatic peritonectomy (DP) for ovarian cancer cytoreduction and to assess associated morbidity. METHODS: Retrospective review yielded 56 patients who underwent DP as part of a cytoreductive procedure for primary or recurrent ovarian cancer between 1988 and 2004. Patients who underwent diaphragmatic resection, removal of diaphragmatic implants with CUSA, cautery, curette, or finger fracture, and patients with pseudomyxoma were excluded from analysis. RESULTS: DP was performed as a component of primary or secondary cytoreduction in 37 (66%) and 19 (34%) patients, respectively. Extended procedures including bowel resection, hepatic resection, splenectomy, or radical hysterectomy were performed with DP in 47 patients (82%). Resection of all disease >1 cm was achieved in 95% (microscopic residuum in 43%). For those undergoing primary cytoreduction, median survival was 59 months and 5-year survival was 49% with median follow-up of 34 months. When performed for recurrent ovarian carcinoma, 5-year survival was 16% and median survival was 23 months. No intra-operative complications could be specifically attributed to DP. Post-operative complications included a 30% rate of pleural effusion which was associated with entry into the pleural space during DP (p<0.0001); thoracentesis was required in 12.5%. CONCLUSIONS: Diaphragmatic metastases are a common obstacle to optimal cytoreduction for patients with ovarian cancer. When necessary, utilizing DP in concert with other extended procedures to obtain maximal cytoreduction is associated with excellent survival. It should be recognized that DP is associated with an increased incidence of post operative pleural effusion, particularly when the pleural space is entered. PMID- 18384867 TI - Antiparkinson drugs used as prophylactics for nerve agents: studies of cognitive side effects in rats. AB - Antiparkinson agents possess excellent anticonvulsant properties against nerve agent-induced seizures by exerting both cholinergic and glutamatergic antagonisms. It is important, however, that drugs used as prophylactics not by themselves cause impairment of cognitive capability. The purpose of the present study was to make a comparative assessment of potential cognitive effects of benactyzine (0.3 mg/kg), biperiden (0.11 mg/kg), caramiphen (10 mg/kg), procyclidine (3 mg/kg), and trihexyphenidyl (0.12 mg/kg) separately and each in combination with physostigmine (0.1 mg/kg). The results showed that benactyzine, caramiphen, and trihexyphenidyl reduced rats' innate preference for novelty, whereas biperiden and procyclidine did not. When benactyzine, caramiphen, and trihexyphenidyl were combined with physostigmine the cognitive impairment disappeared. This counteracting effect, however, caused changes in locomotor and rearing activities not seen by each drug alone. Acetylcholinesterase inhibitors and anticholinergics used as prophylactics can offset each other, but exceptions are observed in a previous study when a very potent anticholinergic (scopolamine) or a high dose of procyclidine still results in cognitive deficits in spite of coadministration with physostigmine. Among the present drugs tested, procyclidine appears to be a robust anticonvulsant with few cognitive side effects. PMID- 18384868 TI - Breast cancer and passive smoking. PMID- 18384869 TI - Marginal and internal fit of metal-ceramic crowns fabricated with a new laser melting technology. AB - OBJECTIVES: The purpose of this in vivo investigation was to evaluate the marginal and internal fit of metal-ceramic crowns fabricated with a new laser melting procedure (BEGO Medical, Bremen, Germany), and to investigate the influence of ceramic firing on the marginal and internal accuracy of these crowns. METHODS: After tooth preparation, impression taking using polyvinylsiloxane and model casting, each preparation was contact less scanned by strip-light-projection. The finishing line and the virtual construction of the metal coping were defined by means of a computer. Using CAD/CAM software the metal copings were produced by BEGO Medical (Germany). A base metal alloy (Wirobond C) and a precious alloy (BioPontoStar, both: BEGO Medical) were used in this study with 14 restorations each. The internal and marginal accuracy of the specimens were examined using a silicone indicator paste (Fit Checker, GC, Japan). After setting, the silicone films were embedded in acrylic resin and sectioned four times. Each slice was photographed digitally at 60x magnification for marginal and at 15x magnification for occlusal adaptation, respectively. Using a light microscope the thickness of the silicone layer was measured at 10 reference points, a total of 3360 measurements. This procedure was repeated after the ceramic firing in the dental laboratory and after intraoral adjustment clinically. RESULTS: No statistically significant differences between the two alloys were found at any time. The mean marginal discrepancies ranged from 74 to 99 microm for both alloys. The internal gaps ranged from 250 to 350 microm. Ceramic firing increased the marginal discrepancies while the internal gaps decreased especially at occlusal surface. However, only in one case a slight statistically significance could be determined (p=0.046). SIGNIFICANCE: The results of this in vivo study show that crowns produced with laser melting technology exhibit a marginal and internal accuracy that is comparable to conventional production procedures. PMID- 18384870 TI - First detection of CYP1A1 and CYP2B induction in Mediterranean cetacean skin biopsies and cultured fibroblasts by Western blot analysis. AB - The need to develop powerful tools to detect exposure and effects of POPs and emerging contaminants in Mediterranean cetaceans led us to develop a suite of sensitive non-lethal biomarkers in integument biopsies of free-ranging animals. In order to propose induction of CYP1A1 and CYP2B, detected by Western blot analysis, as biomarkers of exposure to OCs, PAHs and PBDEs, a three-phase experimental protocol (in vitro experiments, calibration experiments and field applications) was followed using fibroblast cell cultures and biopsies of Mediterranean Stenella coeruleoalba and Tursiops truncatus. This methodology was confirmed to be sensitive and stable in comparison to previous methods used to detect CYP1A1 in biopsies, enabling analysis of several inducible proteins in non lethal samples and analysis of material from stranded animals. PMID- 18384871 TI - Uptake of estradiol from sediment by hornyhead turbot (Pleuronichthys verticalis) and effects on oxidative DNA damage in male gonads. AB - Male and female hornyhead turbot (Pleuronichthys verticalis) were exposed to four concentrations (0, 0.75, 14.7 and 46.5 ng/g dry weight) of E2-amended sediment for 7 days. Sediment-derived E2 was bioavailable to the flatfish, though the route of uptake was unclear. A concentration of 46.5 ng/g E2 in sediment led to a significant increase in vitellogenin concentrations in the plasma in both sexes after 7 days of exposure. Though plasma E2 concentrations increased significantly in males at sediment E2 concentrations of 0.75 ng/g dry weight and above, a dose dependent increase was not observed. There was also no correlation between sediment E2 concentrations, plasma E2 concentrations, and oxidative DNA damage in male gonads. The results suggest that the DNA damage previously seen in the gonads of feral hornyhead turbot at a sewage outfall is likely not caused by acute exposure to exogenous E2 from sediments. PMID- 18384872 TI - Oxidative damage in rainbow trout caged in a polluted river. AB - Sewage treatment works (STWs) are a common source of chemicals entering into the aquatic environment. In order to assess effects of these effluents on oxidative stress parameters in aquatic organisms, we caged rainbow trout at five sites: upstream, near an STW effluent, and three sites downstream in the river Viskan in western Sweden for 14 days during autumn, 2006. We then measured protein carbonyls in plasma as well as 20S proteosome activity and lipid peroxidation products, i.e. MDA and 4-HNE, in liver samples. Levels of both lipid and protein oxidative damage products were elevated in fish caged near the STW effluent while 20S activity showed no differences. This argues that complex mixtures of chemicals entering into the aquatic environment do have deleterious effects on fish. Additionally, oxidative stress parameters can serve as a biomarker in aquatic organisms. PMID- 18384873 TI - DNA strand breaks in grass shrimp embryos exposed to highway runoff sediments and sediments with coal fly ash. AB - Embryo production was reduced in female grass shrimp exposed to sediments with added coal fly ash and to sediments collected from an estuarine station containing high PAH concentrations due to its proximity to a highway storm drain. Grass shrimp embryos exposed to pore water from the high PAH and high metal sediments showed both reduced hatching and increases in DNA strand breaks (comet assay). Sediments with added coal fly ash had high concentrations of vanadium and selenium which may have contributed to effects similar to those observed with sediments with high PAH. The embryo pore water bioassay (hatching/DNA strand breaks) gave results comparable to those observed for reproduction effects (reduced embryo production/embryo hatching) with female grass shrimp exposed to whole sediment. PMID- 18384874 TI - Seasonal variations of exposure biomarkers, oxidative stress responses and cell damage in the clams, Tapes philippinarum, and mussels, Mytilus galloprovincialis, from Adriatic sea. AB - This work investigated the natural variability of several biomarkers in Tapes philippinarum and Mytilus galloprovincialis, sampled from Northern Adriatic where these organisms are important sentinel species for future environmental impact assessment. Levels of metallothioneins, peroxisomal enzymes and acetylcholinesterase, showed a significant seasonality and marked differences between clams and mussels. Among antioxidant enzymes, catalase and GST decreased during the warmer period, the latter enzyme activity resulting particularly high in clams. The total oxyradical scavenging capacity toward peroxyl radicals decreased in mussels from winter to summer, indicating a prooxidant challenge due to higher seawater temperature and intensity of light irradiance. Lysosomal membrane stability did not exhibit significant seasonal variations, while some variations were observed for DNA damages. Overall results indicated a significant influence of seasonal variability on several biomarkers and species-specific differences which should be considered to discriminate the appearance of anthropogenic disturbance. PMID- 18384875 TI - Central and peripheral alytesin cause short-term anorexigenic effects in neonatal chicks. AB - We studied the effects of alytesin, a natural analogue of bombesin, on appetite related responses and behaviors in neonatal chicks. Chicks responded to both intracerebroventricular (ICV) and peripheral injections of alytesin with short term reduced feed intake. ICV alytesin caused reduced short-term water intake when feed was present, but we determined this effect was secondary to feed intake since an effect on water intake was not detected in feed-restricted alytesin treated chicks. The anorexigenic effect of both ICV and peripheral alytesin may be mediated at the hypothalamus, since all hypothalamic nuclei studied; regio lateralis hypothalami, nucleus dorsomedialis hypothalami, nucleus paraventricularis magnocellularis, nucleus perventricularis hypothalami, nucleus infundibuli hypothalami and the nucleus ventromedialis hypothalami, were activated as evident by increased c-Fos immunoreactivity. Central alytesin did not cause increased behaviors that were unrelated to ingestion and did not cause anxiety-related behavior patterns. Additionally, central alytesin did not affect pecking efficacy. We conclude that both ICV and peripheral alytesin injections induce anorexigenic effects in chicks, and the hypothalamus is involved. While the anorexigenic effects of alytesin and bombesin appear to be conserved across species, the two peptides may differ in other behavioral responses and central mechanisms of action. PMID- 18384876 TI - KIS induces proliferation and the cell cycle progression through the phosphorylation of p27Kip1 in leukemia cells. AB - CEM, MOLT4 and SUP-B15 cells were transduced with lentivirus-mediated siRNA KIS gene. The mRNA expressions of KIS were successfully reduced in all cell lines. On the other hand, the mRNA expressions of p27(Kip1) in CEM, MOLT4 and SUP-B15 cells were not affected by the transduction with siRNA KIS gene. We showed that KIS protein directly interacted with p27(Kip1) protein, and reduction of KIS inhibited the S10 phosphorylation of p27(Kip1) in leukemia cells. On these cells transfected with siRNA KIS, the inhibition of S10 phosphorylation of p27(Kip1) was strongly suppressed cell proliferation in a time-dependent manner. Moreover, the inhibition of S10 phosphorylation of p27(Kip1) increased a significant population in G0/G1 fraction. These data demonstrated that the KIS activity was induced during G0/G1, and it promotes cell cycle progression by phosphorylation of S10 on p27(Kip1). We showed that KIS mRNA expression was increased in primary leukemia specimens (acute myelogenous leukemia (AML); 37, myelodysplastic syndrome (MDS); 72, acute lymphoblastic leukemia (ALL); 23), and the mean ratios of KIS to G3PDH in AML, MDS and ALL specimens were 3.62+/-0.68, 3.27+/-0.73 and 3.17+/-0.58, respectively. Moreover, we found that KIS protein was overexpressed in all 132 adults cases of various leukemias, including 37 AML (8 M1, 12 M2, 2 M3, 7 M4, 8 M5), 72 MDS (42 RAEB-I, 30 REAB-II) and 23 ALL (23 L2). This study demonstrates that the elevated levels of KIS protein in leukemia cells promote the cell cycle progression in leukemia cells. PMID- 18384877 TI - Determination of esterase activity and characterization of cholinesterases in the reef fish Haemulon plumieri. AB - White grunt (Haemulon plumieri) has been proposed by the Mesoamerican Barrier Reef System (MBRS) Synoptic Monitoring Program as a bioindicator species. It is in this sense that the present study has a main goal to evaluate this organism's suitability as an indicator species. Individuals were captured during three seasons at the port of Sisal, Yucatan, Mexico which is located in an area that is considered to be weakly impacted by human activities such as agriculture or industry. Both cholinesterase (ChE) and carboxylesterase (CbE) activities were measured in brain, muscle, liver and eye of sampled individuals. Results indicated that ChE and CbE activities were greatest in the brain (256.3 +/- 43) and in the liver (191 +/- 21), respectively. Furthermore, ChEs detected in brain, liver and muscle were characterized, and results suggested that the acetylcholinesterase (AChE) type was more abundant relative to pseudocholinesterase (BChE) which was rare. In addition, K(m) and V(max) and IC(50) values were calculated from the Michaelis-Menten equation. Finally, an additional experiment in vitro showed a significant decrease in both ChE and CbE activities when different tissues were exposed to model xenobiotics, such as benzo[a]pyrene and Chlorpyrifos. In conclusion, findings from this study confirm the potential suitability of H. plumieri as an organic pollution bioindicator species, and thus of practical use for environmental biomonitoring purposes. PMID- 18384878 TI - A role for E2-2 at the DN3 stage of early thymopoiesis. AB - Roles for the E-proteins E2A and HEB during T lymphocyte development have been well established. Based on our previous observations of counter selection against T cells lacking E2-2, it seemed reasonable to assume that there would be a function also for E2-2 in thymocyte development. Aiming at assigning such a role for E2-2, we analyzed the expression of E2-2, E2A, HEB as well as Id mRNA during T cell development. Interestingly, whereas all three E-proteins were expressed during early thymocyte development, significant expression beyond the DP stage was detected only for E2A. Among the Id proteins, Id2 displayed a prominent expression exclusively in DN1, whereas Id3 showed some expression in DN1, followed by a down regulation and then a prominent induction, peaking in the DP stage. E2-2 was expressed during the DN stages, as well as in the DP stage, suggesting that E2-2 operates in concert with the other E-proteins during early thymocyte development. We found that E2-2 null thymocytes displayed a partial block at the DN3 stage of development, as well as a reduced expression of pre-T alpha, known to be regulated also by E2A and HEB. The fact that E2-2 deficient thymocytes develop without gross abnormalities is likely to stem from redundancy due to the co-expression of E2A and HEB. PMID- 18384879 TI - Increased release of sMD-2 during human endotoxemia and sepsis: a role for endothelial cells. AB - MD-2 is the crucial cofactor of TLR4 in the detection of LPS. Here, we show that soluble MD-2 (sMD-2) circulates in plasma of healthy individuals as a polymeric protein. The total amount of sMD-2 in septic plasma was strongly elevated and contained both sMD-2 polymers and monomers, the latter representing the putative biologically active form of MD-2. Moreover, during experimental human endotoxemia, the monomeric and total sMD-2 content in plasma increased with the kinetics of an acute phase protein. The increase in sMD-2 monomers was paralleled by enhanced TLR4 costimulatory activity. The presence of functional sMD-2 during endotoxemia and sepsis was confirmed by immunodepletion. Immunohistochemistry revealed that MD-2 expression in septic patients was strongly enhanced on endothelium and multiple inflammatory cells in lung and liver. In vitro studies showed that sMD-2 release appears to be restricted to endothelial cells and dendritic cells. Release of sMD-2 by endothelial cells was strongly enhanced by LPS and TNF-alpha stimulation. Taken together, this study demonstrates the increase of both circulating polymeric and functional monomeric sMD-2 during endotoxemia and sepsis, and evidence is provided that the endothelium is involved in this process. PMID- 18384880 TI - Liver stroma enhances activation of TLR3-triggered NK cells through fibronectin. AB - Recent studies demonstrate that tissue microenvironments may regulate development and functions of immune cells including dendritic cells, monocytes and T cells. However, there is no report about functional regulation of innate NK cells by tissue microenvironment. The liver is an organ abundant in NK cells and susceptible to virus infection. The number and cytotoxicity of liver NK cells have been shown to be increased during pathogenesis of viral hepatitis and contribute to liver damage. So, whether and how liver stromal microenvironment regulates NK cells need to be fully investigated. By preparing liver fibroblast stromal cells to mimic stromal microenvironment in liver, in this study we demonstrate that liver stroma could chemoattract and adhere TLR3-triggered NK cells, and further augment the cytotoxicity and IFN-gamma production of TLR3 triggered NK cells via fibronectin. Furthermore, the autocrined IFN-gamma from NK cells is required for the enhancement of TLR3-triggered NK cell activation by liver stroma. Our results suggest that liver stroma can recruit NK cells and promote activation of NK cells during viral infection, thus providing a mechanistic explanation for the increased number and cytotoxicity of liver NK cells which may cause liver damage during pathogenesis of viral hepatitis. PMID- 18384881 TI - Continuous-flow complete-mixing system for assessing the effects of environmental factors on colony-level coral metabolism. AB - A small-scale chamber experimental system was designed to study the effects of temperature on colony-level coral metabolism. The system continuously supplies fresh seawater to the chamber, where it is mixed immediately and completely with the seawater already present. This continuous-flow complete-mixing system (CFCM system), in conjunction with theoretical equations, allows quantitative determination of chemical uptake and release rates by coral under controlled environmental conditions. We used the massive hermatypic coral Goniastrea aspera to examine variations in pH, total alkalinity, and total inorganic carbon for 16 days at 27 degrees C under controlled light intensities (300 and 0 micromol m(-2) s(-1)). We confirmed the stability of the CFCM system with respect to coral photosynthetic and calcification fluxes. In addition, we obtained daily photosynthetic and calcification rates at different temperatures (27 degrees C, 29 degrees C, 31 degrees C, and 33 degrees C). When seawater temperature was raised from 31 degrees C to 33 degrees C, the gross primary production rate (Pgross) decreased 29.5%, and the calcification rate (G) decreased 85.7% within 2 days. The CFCM system allows quantitative evaluation of coral colony chemical release and uptake rates, and metabolism. PMID- 18384882 TI - Neuroticism as a mediator of treatment response to SSRIs in major depressive disorder. AB - BACKGROUND: Serotonin function has been implicated in both major depressive disorder and neuroticism. In the current investigation, we examined the hypothesis that any change in depression severity is mediated through the reduction of neuroticism, but only for those compounds which target serotonin receptors. METHODS: Ninety-three outpatients in the midst of a major depressive episode received one of three antidepressant medications, classified into two broad types: selective serotonin reuptake inhibitors (SSRIs) and non-SSRIs (i.e. reversible monoamine oxidase inhibitors [RIMAs] and noradrenergic and dopaminergic reuptake blockers [NDMs]). Patients completed the Hamilton Rating Scale for Depression, Beck Depression Inventory II and Revised NEO Personality Inventory prior to and following approximately 16 weeks of treatment. Structural equation modeling was used to test two models: a mediation model, in which neuroticism change is the mechanism by which SSRIs exert a therapeutic effect upon depressive symptoms, and a complication model, in which neuroticism change is a mere epiphenomenon of depression reduction in response to SSRIs. RESULTS: The mediation model provided a good fit to the data; the complication model did not. Patients treated with SSRIs demonstrated greater neuroticism change than those treated with non-SSRIs, and greater neuroticism change was associated with greater depressive symptom change. These effects held for both self-reported and clinician-rated depressive symptom severity. LIMITATIONS: Replication within a randomized control trial with multiple assessment periods is required. CONCLUSION: Neuroticism mediates changes in depression in response to treatment with SSRIs, such that any treatment effect of SSRIs occurs through neuroticism reduction. PMID- 18384883 TI - Cytokine mRNA expression and pathological findings in pigs inoculated with African swine fever virus (E-70) deleted on A238L. AB - African swine fever virus (ASFV) induces a variety of immune responses and clinical forms in domestic pigs. As it is the only member of the Asfarviridae family, ASFV encodes many novel genes not encoded by other virus families. Among these genes, A238L may regulate the synthesis of pro-inflammatory cytokines, controlled mainly by NFkappaB and NFAT pathways. In this study, we inoculated two groups of pigs, one with the ASFV highly virulent E-70 isolate, deleted on A238L gene, and the other group with the parental E-70 isolate. No significant differences were observed in the clinical signs or pathology between both groups. However, the TNF-alpha mRNA expression was strongly enhanced in the PBMC from pigs inoculated with the virus deleted in A238L, reinforcing the role of the A238L gene in the inhibition of the NFkappaB pathway of expression of cytokines. No up-regulation of pro-inflammatory cytokines was observed in the PBMC of animals inoculated with the E-70 isolate, even though apoptosis and haemorrhages were evident and might be related to the presence of bystander monocyte macrophages expressing these cytokines. Other studies using ASFV deleted in other genes inoculated in the natural hosts should be performed to gain further insight into the role of these genes in the pathogenesis of ASF. PMID- 18384884 TI - Normal development and function of dendritic cells in mice lacking IDO-1 expression. AB - Dendritic cells (DCs) have been shown to express the tryptophan catabolizing enzyme indoleamine 2,3-dioxygenase (IDO-1), a protein presently thought to exert dual and possibly contrasting effects on the immune response. Depletion of tryptophan and release of tryptophan catabolites have been shown to exert a tolerogenic influence on T cell responses, while the IDO enzymatic activity has been recently suggested to promote DC maturation. In this report, we have explored the putative role of IDO-1 in regulating DC biology by analyzing DC development and function from IDO-1 deficient mice. In keeping with previous observations, lack of IDO-1 expression was found to affect in vitro DC generation from bone mouse precursors cultured in the presence of GM-CSF. However, change in growth factor (Flt3L) and/or culture conditions (low-adherence vessels) abolished the difference observed between wt (wild type) and IDO-1-deficient, in vitro generated DCs. Moreover, IDO-1-deficient mice displayed a normal DC compartment in vivo, suggesting that IDO-1 does not play a significant role in DC development and function in vivo. Collectively, these observations suggest that despite a possible role for IDO-1 expression in regulating DC differentiation in vitro under commonly used culture conditions, IDO-1 is largely dispensable for DC development and function in vivo. PMID- 18384885 TI - Human and murine model cell lines for dendritic cell biology evaluated. AB - Dendritic cells (DCs) are specialized antigen presenting cells that link innate and adaptive immune responses. As key mediators of T cell dependent immunity, DCs are considered primary targets for initiating immune responses in infectious diseases and cancer. Conversely, DCs can also play an important role in the induction of tolerance in organ transplantation, autoimmune disorders and allergy. While DCs have been used in clinical trials worldwide during the past decade, many of the highly specialized cell biological characteristics of DCs remain poorly understood. Small numbers of DCs can be isolated as terminally differentiated, post-mitotic cells form either blood or spleen. Alternatively, DC precursors, such as monocytes or bone marrow-derived stem cells, can be isolated and differentiated into DCs in vitro. The relative low numbers of cells that can thus be obtained, combined with difficulties manipulating these terminally differentiated primary cells in vitro and in vivo, have seriously hampered studies aimed at exploring the cell biology of DCs. Good model cell lines therefore provide invaluable tools to study DC biology. So far most DC models are myeloid leukemia-derived cell lines that can be differentiated in vitro towards a DC phenotype. Here, we compared the phenotypical and functional characteristics of frequently used mouse and human DC-model cell lines. We conclude that, although none of these cell lines fully recapitulates all cell biological or immunological features of primary DCs, some of these cell lines provide valuable tools to study specific aspects of DC biology. PMID- 18384886 TI - Clinical fluctuations in MuSK myasthenia gravis are related to antigen-specific IgG4 instead of IgG1. AB - We studied the longitudinal relation between disease severity and titers of antigen-specific IgG subclasses in sera of patients with myasthenia gravis and antibodies to Muscle Specific Kinase (MuSK MG). Six patients were included of whom 55 samples had been collected during 2.5-13.4 years. Anti-MuSK antibodies were determined by ELISA and with a cell-based immunofluorescence assay. Disease severity was scored on a semi continuous scale. Only antigen-specific IgG4, and not IgG1, titers were significantly associated with disease severity in a linear mixed effect model (p = 0.036). Levels of IgG4 antibodies were above IgG1 in all samples except in one patient who went into clinical remission while switching from IgG4 to IgG1. The results support an important role for IgG4 in the pathogenesis of MuSK MG, in contrast to MG with anti-acetylcholine receptor antibodies. PMID- 18384887 TI - Early and later vocalization skills in children with and without cleft palate. AB - OBJECTIVE: The purpose of this study was to describe the early vocalization skills in children with cleft lip and palate (CLP) at 6 and 12 months of age and compare these early vocalization measures to later speech and vocabulary development at 30 months of age. METHODS: The participants in the study included 13 children without cleft lip or palate (NCLP) who were typically developing and 13 children with CLP matched for age, gender and socioeconomic status. Standardized measures of cognition, language, hearing, and prelinguistic vocalization measures were administered at 6 and 12 months and speech production, and vocabulary measures were collected at 30 months of age. RESULTS: Group differences were observed in both receptive and expressive language development at 12 and 30 months of age. Group differences were observed in the frequency of babbling and Mean Babbling Level at 12 months and speech sound accuracy and vocabulary production at 30 months of age. Significant correlation coefficients were observed between babbling frequency at 6 months and consonant inventory size, vocabulary at 30 months for the children with clefts and PCC-R for noncleft children. CONCLUSIONS: This study documented that young children with clefts have persistent vocalization and vocabulary deficits well beyond palate closure. Measures of babbling frequency, Mean Babbling Level and consonant inventories provide clinically effective means of identifying these early deficits. Additionally, these measures may provide a tool for monitoring the effects of early intervention programs that promote facilitation of sound and vocabulary development. PMID- 18384888 TI - Speech and language development in toddlers with and without cleft palate. AB - OBJECTIVE: The effect of early palate closure on speech and language development in children with cleft palate. DESIGN: Comparative study. SETTING: University Medical Center Groningen, Cleft Palate Team (The Netherlands). MATERIALS AND METHODS: Forty-three toddlers with cleft palate and thirty-two toddlers without cleft palate were analyzed with standardized tests for language comprehension and language production. Moreover articulation and hyper nasality were examined by trained speech therapists. RESULTS: For language comprehension, language production and articulation there were no significant differences between the children with and without cleft lip and/or palate. This is despite the high percentage of conductive hearing loss (55%) in children with clefts. Significant difference was found for hyper nasality (mean: 35% vs. 0%, p=0.001). In both groups articulation problems raise to a higher percentage than language production problems (63-20%; 24-4%). CONCLUSIONS: Early surgical treatment is effective for a part of the communicative development, i.e. language development and articulation. Besides conductive hearing loss hyper nasality remains a serious problem in 30-50% of the children with cleft palate. Therefore, speech therapy and pharyngoplasty also are part of the treatment procedure. Because of the high amount articulation problems in all children, standards for articulation development are perhaps too strict. Future research should be carried out after normal variations in articulation development. PMID- 18384889 TI - Beyond NMDA and AMPA glutamate receptors: emerging mechanisms for ionic imbalance and cell death in stroke. AB - The glutamate receptor was one of the most intensely investigated targets for neuroprotection. However, numerous clinical trials of glutamate receptor antagonists for the treatment of stroke were unsuccessful. These failures have led to pessimism in the field. But recent advances could provide hope for the future. This minireview looks beyond the traditional mechanism of glutamate receptor-driven excitotoxicity to identify other mechanisms of ionic imbalance. These advances include findings implicating sodium-calcium exchangers, hemichannels, volume-regulated anion channels, acid-sensing channels, transient receptor potential channels, nonselective cation channels and signaling cascades that mediate crosstalk between redundant pathways of cell death. Further in vivo validation of these pathways could ultimately lead us to new therapeutic targets for stroke, trauma and neurodegeneration. PMID- 18384890 TI - How and why do GPCRs dimerize? AB - Dimerization is fairly common in the G-protein-coupled receptor (GPCR) superfamily. First attempts to rationalize this phenomenon gave rise to an idea that two receptors in a dimer could be necessary to bind a single molecule of G protein or arrestin. Although GPCRs, G proteins and arrestins were crystallized only in their inactive conformations (in which they do not interact), the structures appeared temptingly compatible with this beautiful model. However, it did not survive the rigors of experimental testing: several recent studies unambiguously demonstrated that one receptor molecule is sufficient to activate a G protein and bind arrestin. Thus, to figure out the biological role of receptor self-association we must focus on other functions of GPCRs at different stages of their functional cycle. PMID- 18384891 TI - Chronic stress-induced cellular changes in the medial prefrontal cortex and their potential clinical implications: does hemisphere location matter? AB - The prefrontal cortex (PFC) is implicated in a number of higher cognitive functions as well as processing emotions and regulation of stress responses. Hemispheric specialization of the PFC in humans in emotional processing is well documented, and there is evidence that a similar functional lateralization is present in all mammals. Recent findings suggest the possibility of an intrinsic structural hemispheric asymmetry in the rat medial PFC (mPFC). Specifically, interhemispheric differences have been found in the architecture of pyramidal cell apical dendritic trees together with hemispheric asymmetry in cell proliferation including gliogenesis. It is now well established that chronic stress has a profound impact on neural plasticity in a number of corticolimbic structures and affects the etiology, pathophysiology, and therapeutic outcome of most psychiatric disorders. We summarize recent experimental data documenting pronounced dendritic remodeling of pyramidal cells and suppressed gliogenesis in the mPFC of rats subjected to chronic stress or to artificially elevated glucocorticoid levels. The stress affect on these structural elements seems to be hemispheric specific, often abolishing or even reversing natural asymmetries seen at the cellular level. We discuss these preclinical observations with respect to clinical findings that show impaired function, altered lateralization and histopathological changes in the PFC in psychiatric patients. We argue that it is important to define the kinds of structural changes that result from long-term stress exposure because this knowledge will improve the identification of cellular endophenotypes in various psychiatric disorders. PMID- 18384892 TI - Pattern discrimination is affected by entopallial but not by hippocampal lesions in zebra finches. AB - We compared the effect of entopallial lesions with that of hippocampal lesions on pattern discrimination in zebra finches. The birds were trained to discriminate a feeder with a dot pattern from feeders with a stripe pattern, all located at the floor of a small aviary. When the birds had learned the discrimination, entopallium or hippocampus was damaged by ibotenic acid injection. Entopallial lesions caused deficits in the discrimination, while hippocampal lesions did not. As shown in previous experiments using the same experimental design with feeders of one colour and without distinct pattern [Watanabe S, Bischof HJ. Effects of hippocampal lesions on acquisition and retention of spatial learning in zebra finches. Behav Brain Res 2004;155:147-52], hippocampal lesions cause strong deficits in spatial orientation. Taken together, our experiments prove that the avian hippocampus is involved in spatial learning, but not in pattern discrimination tasks. PMID- 18384893 TI - Different trans RNA splicing events in bloodstream and procyclic Trypanosoma brucei. AB - Most trypanosomatid genes are transcribed into polycistronic precursor RNAs that are processed into monocistronic mRNAs possessing a 39-nucleotide spliced leader (SL) at their 5'-ends and polyadenylation at their 3'-ends. We show here that precursor RNA derived from a luciferase gene integrated in reverse orientation at the rDNA locus of Trypanosoma brucei is processed into three major SL-containing RNAs in bloodstream cells and a single SL-containing RNA in procyclic RNAs. This difference in trans RNA splicing between bloodstream and procyclic cells is independent of the 5'- and 3'-UTRs flanking the luciferase coding region. Thus, bloodstream cells can recognize some sequences in precursor RNA as a SL addition site that procyclic cells do not. These alternative SL addition sites may be aberrant or they might be utilized to expand the number of gene products from individual genes. Future experiments on endogenous genes will be necessary to examine the latter possibility. PMID- 18384894 TI - Opposite effects of oxytocin on proliferation of osteosarcoma cell lines. AB - Oxytocin stimulates proliferation of human osteoblast-like (hOB) cells and human osteosarcoma cells (SaOS-2). In contrast, oxytocin has also been shown to inhibit proliferation of other cell lines such as breast cancer cells. The aim of the present study was to investigate the effects of different concentrations of oxytocin on cell proliferation in osteosarcoma cell lines of different stages of differentiation: SaOS-2, TE-85, and UMR-106. For this purpose cells were incubated with oxytocin (1-1000 pmol/l). Cell proliferation was measured by [(3)H]thymidine incorporation and a commercially available kit (EZ4U). Incubation with oxytocin during 24 h increased proliferation of SaOS-2 cells significantly (100 pmol/l: p<0.01). In contrast, 24 h of incubation with oxytocin decreased proliferation of TE-85 (100 pmol/l: p<0.01) and UMR-106 cells significantly (100 pmol/l: p<0.01). The effects of oxytocin in SaOS-2 and TE-85, but not in UMR-106 cells, were abolished when the cells were incubated with both oxytocin and an oxytocin antagonist (1-deamino-2-D-Tyr-(Oet)-4-Thr-8-Orn-oxytocin). Instead incubation with the oxytocin antagonist alone decreased proliferation of UMR-106 cells significantly (p<0.001). Thus oxytocin has the capacity to both stimulate and inhibit cell proliferation of osteosarcoma cells. This effect might be dependent on the stage of differentiation of the cancer cells. PMID- 18384895 TI - Stroke from paroxysmal atrial flutter or left ventricular hypertrabeculation/noncompaction, visible only on transesophageal echocardiography. PMID- 18384896 TI - Hemodynamic improvement of acutely decompensated heart failure patients is associated with decreasing levels of NT-proBNP. AB - BACKGROUND: Successful therapy in chronic clinically stable heart failure is reflected by decreasing serum NT-proBNP levels. This study evaluates therapy monitoring by NT-proBNP in comparison to invasively measured hemodynamic parameters in acutely decompensated heart failure patients. METHODS AND RESULTS: In 25 acutely decompensated chronic heart failure patients (NYHA III-IV, Cardiac Index (CI)or=15 mm Hg) changes in NT-proBNP and invasive hemodynamics were compared. Hemodynamic improvement in the first 24 h (CI>or=30% and PCWP139) and when the maximum temperature remained >8.9 degrees C during the period 2-4 weeks before slaughter. PMID- 18384899 TI - The link between motor impairment level and motor imagery ability in children with developmental coordination disorder. AB - The present study examined motor imagery ability in children with developmental coordination disorder (DCD). Specifically, it explored whether children with varying degrees of motor impairment differed in their ability to perform motor imagery tasks. Fourty-two children scoring below the 15th percentile on the Movement ABC were split into two groups--DCD severe (DCD-S), scoring on or below the 5th percentile, and DCD mild (DCD-M), scoring from the 6th to 15th percentiles--and compared to 21 age matched controls. Participants performed two motor imagery tasks--hand (performed without and with specific imagery instructions) and whole-body rotation. The results demonstrated that children in the DCD-S group had a generalized motor imagery deficit in that they were less accurate across tasks than controls (and the DCD-M group on the hand task) and showed little benefit when given specific imagery instructions. The DCD-M group appeared capable of performing simpler motor imagery transformations, but were less successful as task complexity increased. Unlike the DCD-S group, the DCD-M group did show some benefit from specific imagery instructions with increases in accuracy on the hand task. These results suggest that a motor imagery deficit does exist in many children with DCD but that its presentation can vary--factors such as the individual child's level of motor impairment and task complexity appear to be linked to the profile of deficits observed, which could explain the inconsistent findings of previous studies. Although this study lends support to the theory that a deficit in internal modeling is an underlying problem for children with DCD, still more research is required to develop the theory further. PMID- 18384900 TI - Comparative full genome analysis revealed E1: A226V shift in 2007 Indian Chikungunya virus isolates. AB - The resurgence of Chikungunya virus (CHIKV) in the form of unprecedented explosive epidemic after a gap of 32 years in India is a point of major public health concern. In 2007 again there was outbreak in Kerala, India, affecting more than 25,000 cases with many reported mortalities. To understand the molecular basis of this high virulence and its implication in large-scale epidemic, a detailed systematic serological, virological and molecular investigation was undertaken with the epidemic samples of Kerala-2007. The comparative analysis of full genome sequence of Chikungunya virus isolate of 2007 with 2006 revealed three unique substitutions in structural and non-structural genes of 2007 isolate [two in E1 region (V14A and A226V) and one in Nsp1 (M184T)]. Our finding further substantiates the association of A226V shift in E1 gene with evolutionary success possibly due to adaptation in the mosquito vector with progression of epidemic, as observed in Reunion Island. This A226V shift which was absent in all 2006 Indian isolates, is found to be present in the four 2007 isolates, analysed in this study. These unique molecular features of the 2007 isolates with the progression of the epidemic from 2005 to 2007 demonstrate their high evolutionary and epidemic potential and thereby suggesting possible implication in higher magnitude and virulence of this outbreak. PMID- 18384901 TI - Down-regulation of Sclerotinia sclerotiorum gene expression in response to infection with Sclerotinia sclerotiorum debilitation-associated RNA virus. AB - We have previously presented convincing evidence in support of a viral etiology for the debilitation phenotype exhibited by strain Ep-1PN of Sclerotinia sclerotiorum. To explore the possible mechanisms underlying fungal pathogenicity and hyphal growth, potential genes whose expression was down-regulated in Ep-1PN were identified from a cDNA library of the virus-free strain Ep-1PNAa, which is a single ascospore derivative of strain Ep-1PN, using reverse northern blot analysis. A total of 1116 cDNA clones were targeted and, following PCR re amplification, 210 cDNA clones were selected as candidates, of which 16 cDNA clones were subjected to northern blot analysis for further confirmation. The results showed that 12 clones represented genes that were differentially expressed in the virus-free strain compared to the virus-infected one. Of the 210 clones that were sequenced, 150 had non-redundant sequences and of these 92% (138 clones) had significant homology to fungal genes in the databases examined. The remaining 12 clones did not have any matches. The differentially expressed genes represented a broad spectrum of biological functions including carbon and energy metabolism, protein synthesis and transport, signal transduction and stress response. This study provides the first insight into genes differentially expressed between the virus-free strain Ep-1PNAa and the virus-infected strain Ep 1PN. The possible relationships between mycovirus-mediated changes in cellular gene expression and observed phenotypes are discussed. PMID- 18384902 TI - In vivo time-dependent gene expression of cationic lipid-based emulsion as a stable and biocompatible non-viral gene carrier. AB - To make stable and biocompatible non-viral gene carriers for therapeutic gene therapy, we developed a cationic lipid-based emulsion (CLE) prepared by an oil-in water (O/W) emulsion method, wherein squalene oil was used as an oil core and the cationic lipid, 1,2-dioleoyl-sn-glycero-3-trimethylammonium-propane (DOTAP), was employed as an emulsifier. To evaluate in vivo characteristics such as toxicity and time-dependent gene expression, a bioluminescence reporter gene in pCMV-luc plasmid DNA was simply mixed with CLE in aqueous condition, resulting in a CLE/DNA complex. The CLE/DNA complex was optimized to form a compact and stable nano-sized particle by adding different amounts of plasmid DNA, and an optimal cationic lipid-to-DNA (C/D) weight ratio of 4 was identified. Freshly prepared CLE/DNA complex, with a C/D of 4, showed a high transfection efficiency and minimal cytotoxicity in vitro, compared to controls of a liposome (DOTAP)/DNA complex and a branched poly(ethyleneimine) (Mw=25 kDa) (bPEI)/DNA complex, respectively. The in vivo characteristics of the CLE/DNA complex were evaluated after intravenous injection into Balb/c mice. Time-dependent gene expression data in vivo were obtained using a non-invasive, whole animal bioluminescence imaging system. These data showed that the CLE/DNA complex offered prolonged high-level gene expression for 1 week, particularly in the liver and spleen. On the other hand, the controls of DOTAP/DNA complex and bPEI/DNA complex showed a relatively lower gene expression, because of the unstable and toxic properties of the control carriers. Our in vivo gene expression data demonstrate the potential of the CLE/DNA complex as a non-viral gene carrier for in vivo gene delivery. PMID- 18384903 TI - In vivo antitumor activity of camptothecin incorporated in liposomes formulated with an artificial lipid and human serum albumin. AB - Camptothecin (CPT) is a strong antitumor agent, but its use limited by its low solubility and the instability of the active lactone form. To overcome these difficulties, liposomes incorporating CPT (CPT liposomes) were designed and tested. CPT liposomes were formulated by the addition of 3,5 bis(dodecyloxy)benzoic acid (DB) to polyethylene glycol-containing liposomes, and by coating the surface of the liposomes with human serum albumin (HSA, HSA-DB-L). HSA-DB-L successfully entrapped CPT with about 80% efficiency and with a particle size of about 150 nm. HSA-DB-L showed attenuated drug release and storage stability. Pharmacokinetics studies in mice showed that i.v. injection of HSA-DB L (2.5 mg/kg) led to prolonged circulation in the plasma; the area under the curve was 22-fold higher than that of CPT solution. The tumor growth in mice with subcutaneous transplantation of colon 26 tumor cells was significantly inhibited after a single i.v. injection of HSA-DB-L at a dose of 15 mg/kg without any significant body weight loss. HSA-DB-L increased the accumulation of CPT in tumor tissue significantly (9.6-fold) more efficiently than CPT solution 24 h after i.v. injection. These findings suggest that HSA-DB-L could increase the stability and the antitumor effect of CPT. CPT delivery by novel liposome formulations is a potential approach for effective treatment of cancer. PMID- 18384904 TI - Vascular endothelial growth factor promotes proliferation and function of hepatocyte-like cells in embryoid bodies formed from mouse embryonic stem cells. AB - BACKGROUND/AIMS: Embryoid bodies (EBs) formed from embryonic stem cells (ESCs) differentiate into hepatocyte-like cells (HLCs), and are thus thought to be a useful cell source for drug testing and bioartificial liver. The aim of this study was to induce proliferation and function of ESC-derived HLCs in EBs using HLC-endothelial cell interaction. METHODS: EBs were cultured in the presence of vascular endothelial growth factor (VEGF) and/or VEGF receptor (VEGFR) inhibitors. To reproduce HLC-endothelial cell interaction, we overexpressed VEGF in ESC-derived HLCs under the control of Cyp7a1 gene in EBs. RESULTS: VEGF added to the cultured EBs increased the proliferation of ESC-derived endothelial cells, resulting in the promotion of proliferation and function of ESC-derived HLCs. In EBs, the VEGFR2 inhibitor suppressed expression of albumin and endothelial cell marker genes, whereas the inhibitor for both VEGFR1 and VEGFR2 suppressed expression of Cyp7a1 and hepatocyte growth factor (Hgf) genes. Upon exposure to VEGF, the endothelial cells in EBs increased Hgf mRNA expression. Forced VEGF expression in ESC-derived HLCs in EBs induced angiogenesis around the HLCs and resulted in an increase in the amount of HLCs. CONCLUSIONS: VEGF indirectly induces the proliferation and function of ESC-derived HLCs through VEGFR1 and VEGFR2 signaling in endothelial cells. PMID- 18384905 TI - Quantification of liver perfusion with [(15)O]H(2)O-PET and its relationship with glucose metabolism and substrate levels. AB - BACKGROUND/AIMS: Hepatic perfusion plays an important role in liver physiology and disease. This study was undertaken to (a) validate the use of Positron Emission Tomography (PET) and oxygen-15-labeled water ([(15)O]H(2)O) to quantify hepatic and portal perfusion, and (b) examine relationships between portal perfusion and liver glucose and lipid metabolism. METHODS: Liver [(15)O]H(2)O-PET images were obtained in 14 pigs during fasting or hyperinsulinemia. Carotid arterial and portal venous blood were sampled for [(15)O]H(2)O activity; Doppler ultrasonography was used invasively as the reference method. A single arterial input compartment model was developed to estimate portal tracer kinetics and liver perfusion. Endogenous glucose production (EGP) and insulin-mediated whole body glucose uptake (wbGU) were determined by standard methods. RESULTS: Hepatic arterial and portal venous perfusions were 0.15+/-0.07 and 1.11+/-0.34 ml/min/ml of tissue, respectively. The agreement between ultrasonography and [(15)O]H(2)O PET was good for total and portal liver perfusion, and poor for arterial perfusion. Portal perfusion was correlated with EGP (r=or+0.62, p=0.03), triglyceride (r=or+0.66, p=0.01), free fatty acid levels (r=or+0.76, p=0.003), and plasma lactate levels (r=or-0.81, p=0.0009). CONCLUSIONS: Estimates of liver perfusion by [(15)O]H(2)O-PET compared well with those by ultrasonography. The method allowed to predict portal tracer concentrations which is essential in human studies. Portal perfusion may affect liver nutrient handling. PMID- 18384907 TI - Pharmaceutical companies and Italian Regional Governments: managing relationships in an increasing institutional complexity. AB - In Italy, the process of power decentralization to Regional Governments has particularly affected pharmaceutical care policies. Regions are experimenting with various strategies to govern drugs utilization and expenditure, and differentiating their approaches, leading to an ever-changing and complex institutional scenario. Pharmaceutical companies have created new professional roles, the Regional Affairs Managers (RAM), with the mandate to monitor the different regional contexts and measures, and to establish relationships with the public actors in charge of pharmaceutical policies. This analysis shows how public affairs/lobbying actions at regional level and the creation of a solid political competence within companies are still in an early phase. The activities carried out by RAMs remain limited to an exchange of information and only rarely are perceived by Regional public servants (RRs) as giving support to their work or influence decisions. The interaction with RAMs is often seen as little relevant and still too concentrated on products and a marketing/commercial approach rather than on broader issues of interest to RRs who need to manage the pharmaceutical care system at large. The level of acceptance of this type of activity is also variable and RRs' attitudes alternate between diffidence, polite tolerance, and openness to a constructive dialogue about pharmaceuticals and their management in a regional healthcare system. PMID- 18384906 TI - Peripheral virus-specific T-cell interleukin-10 responses develop early in acute hepatitis C infection and become dominant in chronic hepatitis. AB - BACKGROUND/AIMS: Interleukin-10 (IL-10) has been ascribed pro-viral but anti fibrotic properties in chronic hepatitis C virus (HCV) infection. In this study, we examined the role of HCV-specific T-cell IL-10 response in patients with acute and chronic HCV infection. METHODS: Peripheral HCV-specific T-cell IL-10 and IFNgamma responses were measured in cytokine Elispot assay using overlapping HCV derived peptides in patients with chronic (n=61), resolved (n=15) and acute (n=8) hepatitis C, looking for their onset, quantity, breadth and durability relative to clinical and virological outcomes. The source and effect of HCV-specific IL-10 response were determined in depletion and IL-10 neutralization experiments. RESULTS: Both HCV-specific IL-10 and IFNgamma responses were detected early within 1-2 months of acute clinical hepatitis C. However, only HCV-specific IL-10 response correlated with elevated liver enzymes, increased viremia and suppressed HCV-specific CD4(+) T-cell proliferation in acute infection. While these associations were lost in established chronic infection, HCV-specific IL-10 responses were increased in patients without cirrhosis while IL-10 blockade enhanced antiviral effector IFNgamma responses. CONCLUSIONS: HCV-specific IL-10 Tr1 responses may play a dual role in HCV infection, dampening effector T-cells to promote viral persistence in acute infection but also protecting against progressive fibrosis in chronic infection. PMID- 18384908 TI - Early evolution of eukaryotic DNA-dependent RNA polymerases. AB - Eukaryotic DNA-dependent RNA polymerases (Pol I-III) share a conserved core of 12 subunits, which is closely related to archaeal RNA polymerases. Rpb8, a subunit found in Pol I, II and III, was thought to be restricted to eukaryotes. We show here that Rpb8 closely resembles an archaeal protein called G, found only in Crenarchaea, which identifies a last missing link between the core structure of archaeal and eukaryotic RNA polymerases. PMID- 18384909 TI - To test or not to test? An evidence-based assessment of the value of screening and monitoring tests when using systemic biologic agents to treat psoriasis. AB - The development of new treatments for psoriasis provides dermatologists novel ways to help control the disease but raises questions about what laboratory screening tests are required. As of yet, no consensus or guidelines exist for dermatologists to follow and there may be misconceptions about the relative need for screening and monitoring tests in patients treated with biologic agents. Current practice ranges from no testing to blanket screening panels. The purposes of this review are to (1) systematically review the literature on the use of screening and monitoring tests when initiating and continuing biologic treatments (adalimumab, alefacept, efalizumab, etanercept, infliximab) for moderate to severe psoriasis or psoriatic arthritis; and (2) suggest practical guidelines for dermatologists on which to base such testing. We searched the Cochrane Collaborative Database (including the Cochrane Database of Systematic Reviews [Cochrane Reviews] and the Cochrane Central Register of Controlled Trials [Clinical Trials]) and the MEDLINE database using medical subject headings as search terms when available or key words when appropriate. We compiled published data on risk and risk assessment related to systemic psoriasis treatments, used expert opinion where appropriate when published clinical data were not adequately informative, and assigned evidence grades for various screening tests based on standard methods of the US Preventive Services Task Force. Finally, we developed a table of evidence grades for tests used to monitor different systemic medications. There is not strong evidence to recommend most screening tests for monitoring biological treatments. Neither is there strong evidence not to do such testing. Ultimately, from a practical standpoint, it is incumbent on the clinician to consider each patient independently and determine what screening tests are most appropriate for each individual patient. PMID- 18384910 TI - Magazine adverts for healthy and less healthy foods: effects on recall but not hunger or food choice by pre-adolescent children. AB - The marketing of foods to children has been criticised by parents and academics alike and the control of such advertising is being considered by politicians. Much of the current research focuses on TV advertising. This study aimed to investigate the effects of exposure to printed advertisements for healthy, less healthy and non-food products on children's mood, hunger, food choice and product recall. Accordingly, 309 children (mean age 9.7 years) received booklets in a quasi-random order. Each booklet contained one of the three types of adverts, ratings of current self-perception and a food choice measure. The booklets were presented as a school-based media literacy exercise. Body weight, height and body satisfaction were assessed 1 week later. The three groups did not differ in the effect on current state or end of session food choice. However, children recalled more of the less healthy food products, even when accounting for recent exposure. Greater product recall of less healthy foods is relevant to future consumption but has a number of possible interpretations. The further exploration of non-TV food marketing is warranted at a time when marketing through these channels is increasing, not least as a result of greater TV advertising regulations. PMID- 18384911 TI - Extended dating couples decision-making strategies about eating: a comparison to married couples. AB - Eating is an important component to the courtship stage of relationships. Therefore, the purpose of the current experiment was to identify common decision making strategies of individuals in extended dating relationships. By doing so, this study was a replication and extension of previous findings by Bonds-Raacke [Bonds-Raacke, J. M. (2006). Using cluster analysis to examine husband-wife decision-making. The Psychological Record, 56 (4), 521-550] on married couples decisions related to eating. Participants for the current study were presented with restaurant menus and asked to rate how likely they would be to eat at each restaurant. The task was completed first independently and then jointly. Results indicated that couples in extended dating relationships employed five common decision-making strategies when jointly selecting a restaurant. Finally, the results revealed that more common decision-making strategies were present among dating couples than married couples. PMID- 18384912 TI - Inverting the food pyramid? Social and cultural acceptability of Walter Willett's dietary recommendations among people with weight concerns. AB - The article presents results from a survey that was carried out among participants in a strictly controlled dietary intervention trial in order to investigate and compare the social and cultural acceptability of three different diets. Measures of social and cultural acceptability included liking of diet, social eating events, practical matters surrounding shopping, cooking, eating, understandings of the relationship between diet type, bodyweight and health, and preferences for specific foods. The survey study focuses especially on the acceptability of the diet recommended by American epidemiologist Walter Willett. On most measures the results indicated that a diet based on Willett's recommendations had a generally high level of acceptability. Scepticism related primarily to the health and weight benefits of this diet in comparison with those of the present dietary recommendations in Denmark. The survey also revealed that participants attributed more influence on their body weight to the amount of food they ate than they did to the composition of the diets they followed. While the scope of the study does not allow for the generalizations of results to a general population level, the experimental design provides detailed insight into social and cultural aspects of experiences of strict dietary adherence. PMID- 18384913 TI - To review or not to review: that is the question. PMID- 18384915 TI - The antihypertensive effect of peptides: a novel alternative to drugs? AB - Many types of bioactive peptides that inhibit angiotensin I, angiotensin I converting enzyme (ACE) and Ang II type 1 receptor (AT1) in the cardiovascular system contribute to the prevention and treatment of hypertension. These inhibitory peptides are derived from many food proteins or artificial synthetic products. Further research examining the bioavailability of ACE inhibitory peptides will lead to the development of more effective ACE inhibitory peptides and foods. Our research also demonstrates that ACE inhibitory peptide LAP may lower blood pressure with no adverse effects. PMID- 18384916 TI - New derivatives of 3,5-substituted-1,4,2-dioxazoles: synthesis and activity against Entamoeba histolytica. AB - Dioxazole derivatives (1-33) were synthesized in two steps via their corresponding oximes (I-III). All the compounds were characterized using various spectroscopic techniques. A comparative study of in vitro antiamoebic activity of these heterocyclic compounds, viz. 3-o-chloro (1-11), 3-m-chloro (12-22) and 3-p chloro (23-33) dioxazoles having same substituents at 5-position of dioxazole ring, was performed against HM1:IMSS strain of Entamoeba histolytica. The results showed a regular pattern in the activity and out of 33 compounds assayed 15 compounds showed better antiamoebic activity than metronidazole with IC(50) values in the range 0.41-1.73 microM and 1.80 microM. Dioxazoles having o-chloro, m-chloro, p-chloro, dichloro and pyridine substituents at 5-position were more active than the standard drug metronidazole. The toxicity studies against human kidney epithelial cell line showed that all the compounds were non-toxic. 3,5-Bis [2-chlorophenyl]-1,4,2-dioxazole (10) was most active and least toxic among all the compounds. PMID- 18384917 TI - Developing an E-learning education programme for staff nurses: processes and outcomes. AB - OBJECTIVE: To describe the Instructional System Design Processes (analysis, design, development, implementation, and evaluation) used to develop an e learning education programme for staff nurses at a 700-bed teaching medical center in Taiwan. BACKGROUND: Evidence has shown that education via e-learning improves knowledge and skills; however, with cost constraints and limited resources, finding effective alternatives is critical for health professionals. METHODS: Five courses required for N2 to N3 level nurses were designed for both e learning and traditional in-classroom programmes. Multiple-choice written test questions and a satisfaction questionnaire were used to compare outcomes between programmes. Nursing care skills were rated as pass or fail. Both programmes were begun at the same time and completed within a 3-month period. RESULTS: Forty-two staff nurses participated in this study. All nurses' knowledge scores were >70 points (out of 100) and all passed nursing care skills. In five courses, only teaching and learning and communication were statistically significantly different (p=0.001). Nearly all nurses (97.6%) felt satisfied or very satisfied with the programmes. CONCLUSIONS: Instructional System Design Processes provide an alternative delivery system for in-service nursing education which allows for individualized leaning. PMID- 18384918 TI - Pox, patient, and plaque. PMID- 18384919 TI - Protection of chickens against avian influenza with non-replicating adenovirus vectored vaccine. AB - Protective immunity against avian influenza (AI) virus was elicited in chickens by single-dose vaccination with a replication competent adenovirus (RCA)-free human adenovirus (Ad) vector encoding an H7 AI hemagglutinin (AdChNY94.H7). Chickens vaccinated in ovo with an Ad vector encoding an AI H5 (AdTW68.H5) previously described, which were subsequently vaccinated intramuscularly with AdChNY94.H7 post-hatch, responded with robust antibody titers against both the H5 and H7 AI proteins. Antibody responses to Ad vector in ovo vaccination follow a dose-response kinetic. The use of a synthetic AI H5 gene codon optimized to match the chicken cell tRNA pool was more potent than the cognate H5 gene. The use of Ad-vectored vaccines to increase resistance of chicken populations against multiple AI strains could reduce the risk of an avian-originating influenza pandemic in humans. PMID- 18384920 TI - Design and evaluation of a community participatory, birth preparedness project in Cambodia. AB - OBJECTIVE: to evaluate a pilot project, which used a community participatory approach to introduce birth preparedness in rural Cambodia. DESIGN: a feasibility and outcome evaluation. This included observation, interview, document analysis and costing. SETTING: the project was undertaken in 15 villages linked to five health centres in Kampong Chhnang, Cambodia. PARTICIPANTS: key management personnel, local midwives, village leaders, village volunteers and village members who had participated in the programme. FINDINGS: community engagement was not only feasible but was also a successful and cost-effective way to introduce birth preparedness. A high degree of satisfaction was reported by the health staff and the community. Over the year in which the project was undertaken, there was a 22% increase in antenatal care, a 32% increase in the number of women delivered by a midwife, and a 281% increase in referrals to hospital. IMPLICATIONS FOR PRACTICE: discussion about birth preparedness should occur not only with pregnant women but also with the communities that support them. Communities that are poor and isolated are responsive to the health needs of their women as they give birth, and articulate their needs when given the opportunity. Interacting with health staff in a way in which there is shared information can lead to greater utilisation of the health services that they provide. PMID- 18384921 TI - Effects of cadence on energy generation and absorption at lower extremity joints during gait. AB - BACKGROUND: Information regarding kinetic changes associated with walking speed is important for identifying alterations in locomotor disorders caused by pathological processes, as opposed to those arising solely from altered speeds. METHODS: Fourteen healthy subjects were assessed walking at both natural and imposed cadences of 60, 80, and 120 steps/min. A 3D motion analysis system, force platforms, and related software were used to obtain kinematic and kinetic data. Net joint powers were calculated across cycles and the area under the positive and negative phases of the power curves provided the mechanical work generated and absorbed at the hip, knee, and ankle. The relative contributions to the total positive and negative work across the four cadences were calculated for each joint. ANOVAs followed by planned contrasts were used to assess the effects of laterality, joint, and cadence. FINDINGS: Power and mechanical work, as well as the contributions of individual joints to the total energy generated and absorbed, were shown to be influenced by walking cadence, independent of laterality. The ankle, knee, and hip contributions to the total limb generation and absorption at the lowest cadence were 53%, 21%, and 26%, and at the highest cadence, the corresponding values were 34%, 33%, and 33%, respectively. INTERPRETATION: Power and mechanical work, as well as the contributions of individual joints to the total energy generated and absorbed, were shown to be influenced by the walking cadence, independent of laterality. These findings will be helpful for identifying walking strategies and adaptations in populations with gait disorders. PMID- 18384922 TI - Brittle bones, pain and fractures--lay constructions of osteoporosis among Norwegian women attending the Nord-Trondelag Health Study (HUNT). AB - Osteoporosis has been labelled the disease of the 21st century. Over the past couple of centuries there have been various notions of this disease in medicine. In the present medical discourse, the emphasis is on prevention rather than treatment, making osteoporosis into a major risk factor for bone fractures. In Norway, osteoporosis is a particularly prevalent condition, leading to bone mass measurements being included in several large health surveys. In a follow-up study of the second round of the Nord-Trondelag Health Study (HUNT), women aged 55-75 years were invited to participate in focus groups to talk about their experiences in relation to their bone density measurements. Findings from these focus groups show that osteoporosis is perceived as a disease characterized by brittle bones, pain and fractures. The physical appearance of a hunchbacked old woman is a dominant way of portraying the disease. It is mainly perceived as prevalent among women, but evidence that men can get it is provided through the example of a famous male athlete who became osteoporotic. Causal explanations for the disease are dominated by culturally shaped anecdotal evidence wherein medical knowledge has been included. Limits to lay constructions of osteoporosis based on such evidence are discussed. Talking about osteoporosis across generations the women applied different explanations for the condition over time. In doing so, they also showed that they have adopted the morality of the new public health where the individual has control over her health through self-governance. Whereas this was no option for their grandmothers, their grandchildren's generation was seen as one failing to meet their obligations to become healthy citizens. The lay construction of osteoporosis can thus be seen as one that has developed from a situation where osteoporotic persons were perceived as victims of harsh circumstances to one of individual responsibility. PMID- 18384923 TI - Factors that influence body image representations of black Muslim women. AB - Research on the body image perceptions of black women is limited. Although previous body image studies have explored the intersection between race and gender, the influence of religion has been neglected. Guided by a grounded theory framework, the focus of this investigation, conducted in Upstate New York, USA, was to examine the role of race and religion in the body image perceptions of 22 African-American Sunni Muslim women. Analysis of individual interviews revealed that, in contrast to using standard medical guidelines, participants' views about their bodies were largely based on positive images of an earlier body size/shape, social and family expectations and contexts, cultural norms and values, and spirituality and religious beliefs. Although the body image perceptions of black Muslim women were similar to those expressed in previous body image studies with black women, participants expressed the importance of highlighting the spiritual versus physical self by adhering to religious guidelines regarding proper dress and appearance. These findings suggest that religion, race, and gender are all important factors to be considered when conducting body image studies with black women. PMID- 18384924 TI - Revisiting labeling theory and health: a commentary on Aronowitz. PMID- 18384925 TI - A case of transient hallucination with ropinirole augmentation antidepressant in a patient with treatment-resistant depression: Is there differential effect of ropinirole dose on developing psychotic symptom? PMID- 18384926 TI - Bone stress and interfacial sliding analysis of implant designs on an immediately loaded maxillary implant: a non-linear finite element study. AB - OBJECTIVES: This study investigated the surrounding bone stress and the implant bone interfacial sliding of implant designs and implant sizes of immediately loaded implant with maxillary sinus augmentation by using three-dimensional (3D) non-linear finite element (FE) analysis. METHODS: Twenty-four FE models including four implant designs (cylindrical, threaded, stepped and step-thread implants) and three implant dimensions (standard, long and wide threaded implants) with a bonded and three levels of frictional contact of implant-bone interfaces were analyzed. The maxillary model was constructed from computer tomography (CT) images of a human skull and all 3D implant models were created via the computer aided design (CAD) software. RESULTS: The use of threaded implants decreased the bone stress and sliding distance obviously about 30% as compared with non threaded (cylindrical and stepped) implants. Increasing the implant's length or diameter reduced the bone stress by 13-26%. Employing a immediately loaded implant with smooth machine surface (mu=0.3, mu represents frictional coefficient) increased the bone stress by 28-63% as compared with the osseointegrated (bonded interfaces) implants. Roughening the implant surface (mu>0.3) did not reduced the bone stress, however it did decrease the interfacial sliding between implant and bone. CONCLUSIONS: For an immediately loaded implant placed with sinus augmentation, using threaded implant could decrease both the bone stress and implant-bone sliding distance which may improve the implant initial stability and long-term survival. Rough surface of implants shows no benefit to reduce the bone stress but they could lower the interfacial sliding. On the contrary, employing long or wide implants decrease the bone stress but they cannot diminish the interfacial sliding. PMID- 18384927 TI - Coverage and current practice patterns regarding assisted reproduction techniques. AB - Access to assisted reproduction techniques (ARTs) is highly variable worldwide. The aim of this paper is to analyse the association between systems for ART coverage, patterns of clinical practice, and outcomes for these procedures. We performed a retrospective analysis of ART activity records in the USA and Europe, and concluded that the availability of ART services, and the activity in this respect, is higher in public than in private coverage systems. Effectiveness decreases with greater coverage, but this occurs in parallel with lower numbers of embryos transferred and with multiple birth rates. Healthcare policy on assisted reproduction should take into account the necessary balance between the effectiveness of the treatment provided and its associated risks, the latter mainly concerning multiple pregnancies. PMID- 18384928 TI - Twin pregnancies in Cardiff and Vale of Glamorgan over four decades: natural twinning is on the increase. AB - OBJECTIVE: To assess trends in twinning over four decades using a population based registry. DESIGN: Ecological study to conduct trend analysis of twin pregnancies in a geographically defined area over 40 years. SETTING: All pregnancies in the Cardiff and Vale of Glamorgan area of South Wales from 1965 to 2004, as recorded in the Cardiff Birth Survey (CBS) database. METHODS: Trends of the incidence of all twin pregnancies (> or = 18 weeks of gestation) were calculated in 5-year increments, beginning with 1965-1969 and ending in 2000 2004. Natural twinning rates could only be calculated for the terminal five time periods (i.e., 1980-1984, 1985-1989, 1990-1994, 1995-1999, and 2000-2004), when information regarding non-spontaneous (iatrogenic) twinning was first collected in the database. All results were adjusted for maternal age. RESULTS: The total twinning rate was 13.1 per 1000 pregnancies in the 1st time period (1965-1969). Subsequently, there was a gradual reduction in twinning, reaching a nadir of 10.3 per 1000 for the time period 1980-1985 (Z=3.15, P value<0.001). This was followed by a gradual increase in twinning, reaching a maximum of 15.7 per 1000 for both 1995-1999 and 2000-2004 (Z=-5.18, P value<0.0001). After exclusion of the cases of iatrogenic pregnancies, the natural twinning rate showed a continuous and gradual increase from 10 per 1000 spontaneous pregnancies in 1980-1984 to 13.3 per 1000 in 2000-2004 (Z=-5.08, P value<0.0001). CONCLUSION: The data showed a gradual, continuous increase in natural twinning rates over the last two decades. Such an increase cannot be attributed to the rise in maternal age alone. PMID- 18384929 TI - Dynamic resolution selection in ultrasonic strain imaging. AB - Ultrasonic strain imaging promises to be a valuable tool in medical diagnostics. Reliability and ease-of-use have become important considerations. These depend on selection of appropriate imaging parameters. Two tasks are undertaken here. The tradeoff between resolution and estimation precision is examined closely to establish models for the relationships with imaging parameters and data properties. These models are then applied in a system that automatically sets the imaging parameters responsive to the data quality and the required estimation precision, so as to produce more meaningful images under varying scan conditions. The new system is applied to simulation, in vitro and in vivo data for validation. It reduces the complexity of the sonographer's role in strain imaging, and produces images of reliable quality even when the level of signal decorrelation varies throughout the ultrasound data. PMID- 18384930 TI - Muscle- and nerve-sparing bulbar urethroplasty: a new technique. AB - BACKGROUND: To describe a new surgical technique for the repair of bulbar urethral strictures to preserve the bulbospongiosum muscle and its perineal innervation. OBJECTIVE: Surgical steps of muscle- and nerve-sparing bulbar urethroplasty are described. The outcome is provided regarding semen sequestration and postvoiding dribbling. DESIGN, SETTING, AND PARTICIPANTS: We performed the procedure in 12 patients (average age: 43.58 yr) with bulbar urethral strictures (average stricture length: 4.47 cm). SURGICAL PROCEDURE: Six patients underwent urethroplasty using a ventral oral mucosal onlay graft, and six patients underwent urethroplasty using a dorsal oral mucosal onlay graft. In all patients, the surgical approach to the bulbar urethra was made avoiding dissection of the bulbospongiosum muscle from the corpus spongiosum and leaving the central tendon of the perineum intact. MEASUREMENTS: Clinical outcome was considered a failure when any postoperative instrumentation was needed. The primary outcome examined the technical feasibility of the muscle- and nerve sparing bulbar urethroplasty. The secondary outcome examined the presence or absence of postoperative postvoid dribbling and semen sequestration using a nonvalidated questionnaire (Appendix). RESULTS AND LIMITATIONS: In all patients, postoperative voiding cystourethrography was performed 3 wk after surgery and no urethral sacculation was evident. Urethrography were repeated after 6 mo and 12 mo. No postvoid dribbling or semen sequestration was demonstrated in all patients at 6 mo and 12 mo after surgery. No patient showed stricture recurrence. The average follow-up was 15.25 mo (range 12 mo to 26 mo, median 13.5 mo). CONCLUSIONS: Bulbar urethroplasty preserving the bulbospongiosum muscle, the central tendon of the perineum, and the perineal nerves is a safe, feasible, minimally invasive alternative to traditional bulbar urethroplasty. PMID- 18384931 TI - On syndromes, symptoms, and root cause. PMID- 18384932 TI - Heterarchy in biological systems: a logic-based dynamical model of abstract biological network derived from time-state-scale re-entrant form. AB - Heterarchical structure is important for understanding robustness and evolvability in a wide variety of levels of biological systems. Although many studies emphasize the heterarchical nature of biological systems, only a few computational representations of heterarchy have been created thus far. We propose here the time-state-scale re-entrant form to address the self-referential property derived from setting heterarchical structure. In this paper, we apply the time-state-scale re-entrant form to abstract self-referential modeling for a functional manifestation of biological network presented by [Tsuda, I., Tadaki, K., 1997. A logic-based dynamical theory for a genesis of biological threshold. BioSystems 42, 45-64]. The numerical results of this system show different intermittent phase transitions and power-law distribution of time spent in activating functional manifestation. The Hierarchically separated time-scales obtained from spectrum analysis imply that the reactions at different levels simultaneously appear in a dynamical system. The results verify the mutual inter relationship between heterarchical structure in biological systems and the self referential property of computational heterarchical systems. PMID- 18384933 TI - Novel signals mediating the functions of somatostatin: the emerging role of NO/cGMP. AB - The neuropeptide somatostatin is a cyclic tetradecapeptide, which is widely distributed in the peripheral and central nervous system. It mediates a plethora of physiological actions and functions as a neurotransmitter, neuromodulator or trophic factor. Somatostatin activates six receptor subtypes that are expressed differentially in different tissues and are coupled to diverse signalling pathways. In order to elucidate the functional role of the individual receptor subtypes, many investigations focused on the assignment of each receptor to a particular signalling pathway. Signalling pathways involving enzyme (adenylate cyclase, phospholipases, phosphatases) and ion channel systems in native and recombinant receptor systems have been extensively studied. A one to one situation (receptor/pathway) has yet to be established, thus justifying the diverse actions of somatostatin. Recently, a NO/cGMP pathway has been shown to mediate the functions of somatostatin and its receptors. This review will present the findings that support the emerging role of NO/cGMP as a novel signal in SRIF's actions in retinal physiology and somatotroph release. PMID- 18384934 TI - Editorial. Supplement issue: basic and pharmacological aspects of cannabinoid activity in nervous and reproduction systems. PMID- 18384935 TI - Clinical analysis for an unusual large cystic meningioma: case report and review of the literature. AB - The authors report the case of a 17-year-old boy with an unusual large cystic meningioma (Nauta type II) in the right hemisphere. The imaging appearances of this patient were very unusual. The shape of the huge cyst was crescentic and similar to subdural hematoma. It lay between the dura and the solid tumor parts. In addition there was a small intracystic nodule attached to the cyst wall. The patient underwent a right hemisphere craniotomy. At surgery it was found that the cyst contained a large amount of xanthochromic fluid and some semitransparent serumlike sediment. The intracystic nodule was proved to be necrotic substance without tissue and cell structure. Histological examination displayed an anaplastic meningioma, of which the cyst wall also consisted of meningioma tissue. To the best of the authors' knowledge, such an unusual case of cystic meningioma has not been reported. The authors review the literature with reference to intratumoral cyst associated with meningiomas, analyze the unusual imaging appearances of this patient, and explore the mechanism of cyst formation. The mechanism of cyst formation associated with meningiomas is not perfectly understood. Intratumoral cyst formation may be attributed to microcystic degeneration, ischemic necrosis, intratumoral hemorrhage, transudation and secretory changes within the tumor. PMID- 18384936 TI - Cerebrospinal fluid rhinorrhea following trans-sphenoidal pituitary macroadenoma surgery: experience from 592 patients. AB - OBJECTIVES: To determine the incidence, risk factors, diagnostic procedures, and management of cerebrospinal fluid (CSF) leaks following trans-sphenoidal pituitary macroadenoma surgery. METHODS: Retrospective analysis of 592 patients. RESULTS: Intra- and post-operative CSF leaks occurred in 14.2 and 4.4% of patients, respectively. Surgical revision, tumor consistency, and tumor margins were independently associated with intra-operative leaks, while the tumor size, consistency, and margins were risk factors of post-operative leaks. The intra operative leak rate of ACTH adenomas was greater than all other types combined; the incidence of post-operative CSF leaks was highest for FSH adenomas. There were no significant differences among various techniques and we achieved an initial repair success rates of 83.3 and 92.9% for intra- and post-operative CSF leaks, respectively. Of the 26 patients with post-operative CSF leaks, five were complicated by meningitis and four by post-infectious hydrocephalus which required ventriculoperitoneal shunts. CONCLUSIONS: CSF leaks have a propensity to occur in cases with fibrous tumors or tumors with indistinct margin and may have some relationship with the tumor type. Endoscopic and microscopic repairs were shown to be effective techniques in managing these types of leaks. Post infectious hydrocephalus may influence the outcome of the repair and ventriculoperitoneal shunts were necessary in some cases. PMID- 18384937 TI - Thyroid fine needle aspiration: how to improve clinicians' confidence and performance with the technique. AB - Studies from single institutions report an acceptable accuracy rate for thyroid fine needle aspiration (FNA). However, FNA accuracy is much lower in many other centers in Europe and the USA and large multicenter studies indicate that the clinicians' confidence in the FNA technique remains low. One explanation for this is that there is an excess of inadequate and indeterminate findings for a follicular nodule at FNA cytology. In a University Hospital with large and qualified experience on thyroid nodule diagnosis, a review of 320 slides with an FNA diagnosis of indeterminate follicular nodule from different minor Italian Hospitals led to a different diagnosis in 61%. Since ancillary thyroid imaging may be overutilized and only a few authors report a proportion of excised nodules lower than 10%, we suspect that use of the FNA procedure is suboptimal. Several techniques are reported to improve the performance of thyroid FNA. Among these are tumor markers and large needle aspiration biopsy (LNAB). Immunodetection of the tumor marker galectin-3 has been evaluated by large multinational studies. Analysis of LNAB specimens reduces the number of inadequate FNA findings, improves the diagnostic determination of indeterminate follicular FNA findings and represents a better substrate for the determination of galectin-3. Therefore, we propose that clinical practice guidelines reflect these adjuvant techniques to thyroid FNA in order to improve selection criteria for thyroid nodule surgery. PMID- 18384938 TI - Dormant tumor cells as a therapeutic target? AB - Tumor dormancy is characterised by the persistence of residual tumor cells for long periods. Recurrence from minimal residual disease is a major cause of cancer death. Thus, understanding how cancer cells become and remain dormant, may lead to new strategies to prevent relapse. Evidence has emerged that a balance exists between host and dormant tumor cells. Cross-talk between tumor cells and their micro-environment, angiogenesis, and anti-tumor immune response participate in the control of dormant tumor cells. Tumor cells have several mechanisms of maintaining equilibrium, and immune escape, including expression of immuno regulatory molecules (e.g., increased expression of B7.1 and B7-H1); epigenetic modifications (e.g., silencing of the SOCS1 gene, de-regulating the JAK/STAT pathway); and autocrine loops. These new findings offer new opportunities to design specific treatments, to modify the balance in favor of the host immune response. PMID- 18384939 TI - ZNF23 induces apoptosis in human ovarian cancer cells. AB - We recently reported that the level of ZNF23, a KRAB-containing zinc finger protein, is reduced in human cancers and it inhibits cell growth by inducing cell cycle arrest. Here we showed that ZNF23 also induces apoptosis in ovarian cancer cells. The protein level of ZNF23 in ovarian cancers was greatly down-regulated compared with that in the normal ovaries. Introduction of ZNF23 into ovarian cancer cells led to apoptosis as demonstrated by activation of caspase-3, nuclear condensation and formation of a sub-G1 peak. This apoptotic process was correlated with loss in mitochondrial membrane potential, cytochrome c release and caspase-9 activation. Furthermore, ZNF23 induced apoptosis partially via down regulation of Bcl-XL. Thus, our results suggest that ZNF23 may also induce apoptosis to suppress tumor cell growth and points to the possibility that its down-regulation might facilitate ovarian cancer cell survival. PMID- 18384941 TI - Prohibitin identified by proteomic analysis of prostate biopsies distinguishes hyperplasia and cancer. AB - Prostate cancer (PCA) is the most common type of cancer found in men of western countries and is the leading cancer death next to lung cancer and colorectal cancer. Prostate-specific antigen (PSA) test is an established diagnostic tool for PCA detection, but confirmation of diagnosis by histopathological evaluation of prostate needle biopsies is performed. To define protein expression pattern of prostate biopsies, in the present study we investigated biopsy samples from benign prostate hyperplasia (BPH, n=11) and prostate cancer (PCA, n=12) patients by two-dimensional gel electrophoresis (2-DE) and mass spectrometry to identify potential biomarkers which might distinguish the two clinical situations. 2-DE results revealed 88 protein spots expressed differentially among hyperplasia and cancer groups with statistical significance. Interesting spots were analyzed by MALDI-TOF-MS-MS and 79 different proteins were identified. The important proteins identified included prostatic acid phosphatase precursor, a significant overexpressed protein in PCA, prohibitin, NDRG1 tumor suppressor proteins, heat shock proteins, cytoskeletal proteins, enzymes like DDAH1 and ALDH2. Prohibitin was investigated in detail at mRNA level and protein level using immunohistochemistry on prostatectomized specimens. We found that the level of mRNA for prohibitin correlates with the increased amount of protein indicating involvement of changes at transcriptional level. Furthermore, immunohistochemistry revealed no staining in BPH (n=13), moderate staining in prostate intra-epithelial neoplasia (PIN, n=5) but strong staining in PCA (n=18). Our results demonstrate that protein profiling and mRNA studies can be performed on the same prostate biopsy. Moreover, our study revealed a significant up regulation of prohibitin in prostate cancer compared to BPH which may be a potential marker to distinguish PCA and BPH. Some of the interesting proteins identified in this approach may serve to develop new targets for PCA diagnosis and treatment. PMID- 18384940 TI - Hypoxia in microscopic tumors. AB - Tumor hypoxia has been commonly observed in a broad spectrum of primary solid malignancies. Hypoxia is associated with tumor progression, increased aggressiveness, enhanced metastatic potential and poor prognosis. Hypoxic tumor cells are resistant to radiotherapy and some forms of chemotherapy. Using an animal model, we recently showed that microscopic tumors less than 1mm diameter were severely hypoxic. In this review, models and techniques for the study of hypoxia in microscopic tumors are discussed. PMID- 18384942 TI - Anticancer drugs and central nervous system: clinical issues for patients and physicians. AB - Anticancer drugs may cause neurological toxicity involving the central nervous system. Patients receiving anticancer treatment may develop encephalopathy, extrapyramidal reactions, seizures, cerebellar dysfunction, retinopathy, cerebral venous thrombosis, myelopathy, cognitive impairment, and psychiatric symptoms. Physician should carefully evaluate neurological signs and symptoms in order to recognize these drug-related adverse reactions. In this review we aimed at presenting different neurological complications of anticancer drugs and their management. PUBMED search was performed in order to retrieve all articles and case reports dealing with central nervous system toxicity related to anticancer treatments. PMID- 18384943 TI - Photocatalytic removal of M(2+) (Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+) and Ag(+)) over new catalyst CuCrO(2). AB - The metal ions M(2+) (Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+) and Ag(+)) are potentially toxic. Their electro deposition has been carried out in aqueous air equilibrated CuCrO(2) suspension upon visible illumination. The delafossite CuCrO(2) is p-type semiconductor characterized by a low band gap (1.28 eV) and a long-term chemical stability. The corrosion rate is found to be 10(-2) micromol m(-2)month(-1) in aqua regia. The oxide has been elaborated through nitrate route where the specific surface area is increased via the surface/bulk ratio. A correlation exists between the dark M(2+) adsorption, the redox potential of M(2+/0) couple and the conduction band of CuCrO(2) positioned at -1.06 V(SCE). Ag(+) cannot be photoreduced because of its positive potential located far above the valence band. By contrast, Zn(2+) is efficiently deposited due to the large driving force at the interface. The improved photoactivity of copper with a deposition percentage (90%) is attributed to the strong dark adsorption onto the surface catalyst. The results indicate a competitive effect with the water reduction; it has been observed that the M(2+) deposition goes parallel with the hydrogen evolution. Such behavior is attributed to the low H(2) over voltage when ultra fine aggregate of M islands are photodeposited onto CuCrO(2) substrate. PMID- 18384944 TI - Interaction of soil, water and TNT during degradation of TNT on contaminated soil using subcritical water. AB - Subcritical water was used at laboratory scale to reveal information with respect to the degradation mechanism of TNT on contaminated soil. Highly contaminated soil (12% TNT) was heated with water at four different temperatures, 150, 175, 200 and 225 degrees C and samples were obtained at appropriate time intervals. At the same time, similar experiments were performed with TNT spiked on to clean soil, sand and pure water in order to compare and eliminate various factors that may be present in the more complex contaminated soil system. Subcritical water was successful at remediating TNT-contaminated soil. TNT destruction percentages ranged between 98 and 100%. The aim of this work was to study the soil-water contaminant interaction and determine the main physical parameters that affect TNT degradation. It was shown that the rate-limiting step of the process is the extraction/diffusion of TNT molecules from the soil core to the soil surface, where they degrade. Additionally, it was determined that the soil matrix also catalyses degradation to a lesser extent. Autocatalytic effects were not clearly observed. PMID- 18384945 TI - Synthesis of crosslinked starch-graft-polyacrylamide-co-sodium xanthate and its performances in wastewater treatment. AB - A novel crosslinked starch-graft-polyacrylamide-co-sodium xanthate (CSAX) was synthesized by grafting copolymerization reaction of corn starch, acrylamide (AM), and sodium xanthate using epichlorohydrin (EPI) as cross-linking reagent and ceric ammonium nitrate (CAN) as initiator in aqueous solution. CSAX was characterized by FTIR and element analysis. The performances of CSAX in wastewater treatment were evaluated by flocculation experiment. The results show that the CSAX was successfully synthesized and it has functions of removing both turbidity and copper ions from aqueous solution. It was proved that CSAX is more effective than crosslinked starch xanthate (CSX) and much more effective than crosslinked starch-graft-polyacrylamide (CSA) for removing copper ions. It was also proved that CSAX is little less effective than CSA, but much more effective than CSX for removing turbidity. PMID- 18384946 TI - Decontamination of solutions containing EDTA using metallic iron. AB - EDTA removal from solutions using metallic iron was carried out at different values of pH, iron load and concentrations at free access of air and in closed vessels. The EDTA destruction was investigated using chemical and capillary electrophoresis analysis. Fe corrosion was studied voltammetrically and the composition of the precipitate formed was investigated using FT-IR spectroscopy and chemical analysis. The EDTA decomposition is remarkably enhanced by the addition of Cu(II) to the EDTA solutions and access of air. The precipitation of the derivatives of insoluble Fe with EDTA or its decomposition products proceeds along with the destruction of EDTA. In closed systems the main EDTA removal reaction is precipitation with iron ions. PMID- 18384947 TI - Kinetics and modified clay thermodynamic from the Brazilian amazon region for lead removal. AB - Modified Brazilian smectite-bearing clay samples displayed ability for lead adsorption. The structure modification of smectite were obtained through pillaring process and functionalization with [3-(2 aminoethylamino)propyl]trimethoxysilane. The chemical modification process increases the basal spacing of the natural smectite from 1.354 to 2.364 nm. The Langmuir model was fitted to experimental data in linear regression. Kinetic studies showed an equilibrium adsorption time of 700 min on the modified clay. The experimental data were correlated with two distinct kinetic models were used: (i) external mass transfer diffusion and (ii) intraparticular mass transfer diffusion. However, the intraparticle mass transfer diffusion model gave a better fit to these experimental data. The energetic effects caused by lead interactions were determined through calorimetric titration at the solid-liquid interface and gave a net thermal effect that enabled the calculation of the exothermic values and the equilibrium constant. PMID- 18384948 TI - Correlation between self-ignition of a dust layer on a hot surface and in baskets in an oven. AB - Evaluation of self-ignition hazard of bulk materials requires experimental determination of self-ignition temperatures as a function of volume. There are two standardised methods: (1) determination of the self-ignition temperature of dust samples in oven and (2) measurement of the self-ignition temperature of a dust layer deposited on a hot surface. Sometimes, the sample behaviour during these tests makes the second method difficult to apply. The self-ignition phenomena in these two tests rely on the same principles. Their results are interpreted with the help of theoretical relations. The correlation described in this paper can be considered acceptable to deduce self-ignition temperature of a dust layer, based on results of self-ignition of the same dust in heating ovens, if the Biot number (alpha) can be estimated. Uncertainty on the correlation is near 30K. This uncertainty is on the same order of magnitude as the difference in the self-ignition temperature on a hot surface for thickness between 5 and 15 mm. PMID- 18384949 TI - Phosphoric acid modified-Y zeolites: a novel, efficient and versatile ion exchanger. AB - Large pore HY zeolite was modified with phosphoric acid by wet method. The modified zeolite was converted to Na(+) form using aqueous NaHCO(3) solution(.) The Na(+) form of modified zeolite, represented as PNa(2)Y, was characterized by XRD, BET surface area, SEM, and AAS techniques. The XRD analysis showed diffraction patterns same as that of parent HY zeolite, as a result there has been no structural degradation during modification. It was then tested for sorption of Cu(2+) ions from aqueous solution. The Cu(2+) content of the solution was analyzed by AAS. PNa(2)-Y shows higher sorption capacity ( approximately 40%) than the parent Na-Y ( approximately 23%) zeolite, which is attributed to the double of amount Na(+) content in PNa(2)-Y compared to the Na-Y zeolite. Equilibrium modeling data were found to fit more to the linear Langmuir model than the Freundlich model. The thermodynamic parameters such as change in free energy (DeltaG degrees ), enthalpy (DeltaH degrees ), and entropy (DeltaS degrees ), were also calculated. These parameters confirmed that the sorption of Cu(2+) is feasible, spontaneous and endothermic. PMID- 18384950 TI - Utilization of flotation wastes of copper slag as raw material in cement production. AB - Copper slag wastes, even if treated via processes such as flotation for metal recovery, still contain heavy metals with hazardous properties posing environmental risks for disposal. This study reports the potential use of flotation waste of a copper slag (FWCS) as iron source in the production of Portland cement clinker. The FWCS appears a suitable raw material as iron source containing >59% Fe(2)O(3) mainly in the form of fayalite (Fe(2)SiO(4)) and magnetite (Fe(3)O(4)). The clinker products obtained using the FWCS from the industrial scale trial operations over a 4-month period were characterised for the conformity of its chemical composition and the physico-mechanical performance of the resultant cement products was evaluated. The data collected for the clinker products produced using an iron ore, which is currently used as the cement raw material were also included for comparison. The results have shown that the chemical compositions of all the clinker products including those of FWCS are typical of a Portland cement clinker. The mechanical performance of the standard mortars prepared from the FWCS clinkers were found to be similar to those from the iron ore clinkers with the desired specifications for the industrial cements e.g. CEM I type cements. Furthermore, the leachability tests (TCLP and SPLP) have revealed that the mortar samples obtained from the FWCS clinkers present no environmental problems while the FWCS could act as the potential source of heavy metal contamination. These findings suggest that flotation wastes of copper slag (FWCS) can be readily utilised as cement raw material due to its availability in large quantities at low cost with the further significant benefits for waste management/environmental practices of the FWCS and the reduced production and processing costs for cement raw materials. PMID- 18384951 TI - Mathematical evaluation of activated carbon adsorption for surfactant recovery in a soil washing process. AB - The performances of various soil washing processes, including surfactant recovery by selective adsorption, were evaluated using a mathematical model for partitioning a target compound and surfactant in water/sorbent system. Phenanthrene was selected as a representative hazardous organic compound and Triton X-100 as a surfactant. Two activated carbons that differed in size (Darco 20-40 mesh and >100 mesh sizes) were used in adsorption experiments. The adsorption isotherms of the chemicals were used in model simulations for various washing scenarios. The optimal process conditions were suggested to minimize the dosage of activated carbon and surfactant and the number of washings. We estimated that the requirement of surfactant could be reduced to 33% of surfactant requirements (from 265 to 86.6g) with a reuse step using 9.1g activated carbon (>100 mesh) to achieve 90% removal of phenanthrene (initially 100mg kg-soil(-1)) with a water/soil ratio of 10. PMID- 18384952 TI - Study on the characteristics of building bricks produced from reservoir sediment. AB - This research investigates the feasibility of building bricks produced from reservoir sediment sintering using various sintering temperatures and clay additions. The experimental results indicate that sintered specimen densification occurred at sintering temperatures of 1050-1100 degrees C. Increasing the sintering temperature decreases the water absorption and increases the shrinkage, density and compressive strength of sintered specimens. The experiments were conducted at a temperature ranged from 1050 to 1150 degrees C with clay addition contents varying from 0% to 20%. All sintered specimens made from reservoir sediment were in compliance with Taiwan building bricks criteria. This means that raw materials for producing building bricks can be replaced with reservoir sediment. The metals concentrations of the leachate from the toxicity characteristics leaching procedure (TCLP) test are all complying with the current regulatory limits. These results confirm the feasibility of using reservoir sediment to produce sintered construction brick. PMID- 18384953 TI - Adsorptive studies of hazardous dye Tropaeoline 000 from an aqueous phase on to coconut-husk. AB - This contribution presents result pertaining to the adsorptive removal of water soluble hazardous dye Tropaeoline 000, on to a low cost adsorbent-coconut-husk and its efficiency in the dye colour sorption was compared with activated carbon (AC). The results obtained from the batch experiments revealed the ability of the coconut-husk to remove the Tropaeoline 000 dye. The adsorption of dye was carried out at different pH, temperatures, amount of adsorbent, contact time, concentration of adsorbate (initial dye concentration) and particle size. The adsorption studies revealed that the ongoing adsorption validates both Langmuir and Freundlich adsorption isotherm at temperatures 30, 40 and 50 degrees C. The adsorption isotherm data was also employed to calculate thermodynamic parameters like DeltaG degrees , DeltaH degrees and DeltaS degrees . The negative values of DeltaG degrees indicate that the dye adsorption process is a spontaneous in nature. The positive value of DeltaH degrees shows the endothermic nature of AC/coconut-husk system whereas negative value of DeltaH degrees indicates that the exothermic nature of AC/coconut-husk. The adsorption was found to undergo via a pseudo-second order adsorption kinetics and the results revealed that coconut husk, an agricultural waste, proved to be an excellent low cost adsorbent. PMID- 18384954 TI - Environmental impacts of waste incineration in a regional system (Emilia Romagna, Italy) evaluated from a life cycle perspective. AB - The advisability of using incineration, among the other technologies in Municipal Solid Waste Management, is still a debated issue. However, technological evolution in the field of waste incineration plants has strongly decreased their environmental impacts in the last years. A description of a regional situation in Northern Italy (Emilia Romagna Region) is here presented, to assess the impacts of incinerators by the application of Life Cycle Assessment (LCA) methodology and to stress the most impacting steps in incineration process. The management of solid residues and heavy metal emission resulted the most important environmental concerns. Furthermore, a tentative comparison with the environmental impact of landfill disposal, for the same amount of waste, pointed out that incineration process must be considered environmentally preferable. PMID- 18384955 TI - Sorption and desorption of Cd, Cr, Cu, Ni, Pb and Zn by a Fibric Histosol and its organo-mineral fraction. AB - It has often been stated that the contribution of soil organic matter (OM) to the sorption of heavy metals can be evaluated using the surface horizon of a Histosol as typical of soil organic matter. However, components of Histosols other than organic matter, such as clay minerals and Fe or Mn oxides, can also sorb heavy metals. In this work we compared the heavy metal sorption and desorption behaviour of a Fibric Histosol H horizon with that of its organo-mineral fraction (OMF, defined as the fraction of wet particle size <100 microm) in experiments in which Cd, Cr, Cu, Ni, Pb and Zn were sorbed simultaneously from solutions of various concentrations. The OMF sorbed the metals reversibly and apparently mainly at specific sites to each particular metal, in keeping with the good fit of Langmuir isotherms to the sorption data; greatest sorption capacity was for lead and copper. Whole H horizon appeared to include sites at which binding was less reversible and chromium competed with the other metals, especially copper. Organo-mineral fraction is suggested to evaluate the soil organic matter contribution to heavy metal fixation. PMID- 18384956 TI - Association of polymorphisms in the haplotype block spanning the alternatively spliced exons of the NTNG1 gene at 1p13.3 with schizophrenia in Japanese populations. AB - Chromosome 1p13 is linked with schizophrenia in Japanese families, and one of the candidate genes in this region is the netrin G1 (NTNG1) gene at 1p13.3. Associations of 56 tag single-nucleotide polymorphisms (SNPs) with schizophrenia were explored by transmission disequilibrium analysis in 160 Japanese trios and by case-control analysis in 2,174 Japanese cases and 2,054 Japanese controls. An association between SNP rs628117 and schizophrenia was identified by case-control comparison (nominal allelic p=0.0009; corrected p=0.006). The associated polymorphism is located in intron 9 and in the haplotype block encompassing the alternatively spliced exons of the gene. Allelic association of a different SNP in the same haplotype block in Japanese families was previously reported. These findings support that the NTNG1 gene is associated with schizophrenia in the Japanese. PMID- 18384957 TI - Post-infarct cardiac sympathetic hyperactivity regulates galanin expression. AB - The neuropeptide galanin is elevated in the cardiac sympathetic innervation after myocardial infarction (MI). Galanin inhibits vagal transmission and may support the regeneration of sympathetic nerves, thereby contributing to the development of arrhythmia and sudden cardiac death after MI. The reason for increased galanin production in sympathetic neurons after myocardial infarction is not known. Cardiac sympathetic neurons are activated chronically after cardiac ischemia reperfusion, and activation of sympathetic neurons in culture stimulates galanin expression. Therefore, we tested the hypothesis that increased sympathetic nerve activity stimulates galanin expression in cardiac sympathetic neurons after myocardial infarction. To test this hypothesis we used TGR(ASrAOGEN) transgenic rats, which lack brain angiotensinogen and do not exhibit post-infarct sympathetic hyperactivity. Hearts and stellate ganglia were collected 1 week after ischemia-reperfusion. Galanin mRNA was quantified by real-time PCR and peptide content was assayed by enzyme-linked immunosorbent assay. Galanin mRNA increased approximately 3-fold after MI in cardiac sympathetic neurons of both genotypes compared to unoperated and sham controls. Left ventricular galanin content, however, increased after MI only in Sprague-Dawley rats and not in AOGEN rats. These data suggest that post-infarct cardiac sympathetic hyperactivity stimulates galanin peptide production but is not required for increased galanin mRNA expression. PMID- 18384958 TI - Brain dynamics for perception of tactile allodynia (touch-induced pain) in postherpetic neuralgia. AB - Postherpetic neuralgia (PHN) is a debilitating chronic pain condition often accompanied by a sensation of pain when the affected region is touched (tactile allodynia). Here we identify brain regions involved in stimulus-induced touch evoked pain (dynamical mechanical allodynia, DMA), compare brain activity between DMA and spontaneous pain (described earlier for the same patients in [Geha PY, Baliki MN, Chialvo DR, Harden RN, Paice JA, Apkarian AV. Brain activity for spontaneous pain of postherpetic neuralgia and its modulation by lidocaine patch therapy. Pain 2007;128:88-100]), delineate regions that specifically code the magnitude of perceived allodynia, and show the transformation of allodynia related information in the brain as a time-evolving network. Eleven PHN patients were studied for DMA and its modulation with Lidoderm therapy (patches of 5% lidocaine applied to the PHN affected body part). Continuous ratings of pain while the affected body part was brushed during fMRI were contrasted with non painful touch when brushing was applied to an equivalent opposite body site, and with fluctuations of a bar observed during scanning, at three sessions relative to Lidoderm treatment. Lidoderm treatment did not decrease DMA ratings but did decrease spontaneous pain. Multiple brain areas showed preferential activity for allodynia. However, mainly responses in the bilateral putamen and left medial temporal gyrus were related to the magnitude of allodynia. Both DMA and spontaneous pain perceptions were best represented within the same sub-cortical structures but with minimal overlap, implying that PHN pain modulates behavioral learning and hedonics. These results have important clinical implications regarding adequate therapy. PMID- 18384959 TI - Long-term trials of pregabalin and duloxetine for fibromyalgia symptoms: how study designs can affect placebo factors. PMID- 18384960 TI - Not so fast: the reformulation of fentanyl and breakthrough chronic non-cancer pain. PMID- 18384961 TI - The sensitivity of PCR and serology in different Theileria parva epidemiological situations in Rwanda. AB - Theileria parva is the causative agent of a lethal tick-borne disease of cattle occurring in eastern, central and southern Africa. Variations in the sensitivity of the serological and molecular tests with seasonal vector occurrence and discrepancies between low PCR prevalence and high T. parva vector density are a setback to estimate true prevalences. Therefore, the objectives of the present studies were to evaluate (1) the sensitivity of three serological tests (IFAT, ELISA and SELISA) and one molecular test (PCR) in the diagnosis of chronic T. parva infections in four different agro-ecological zones of Rwanda and (2) the effect of tick challenge and animal's age on the sensitivity of PCR. Blood samples from 635 bovines were collected in four agro-ecological zones of Rwanda. All sera were screened using the IFAT, ELISA, SELISA and PCR. The binary results of the four diagnostic tests were introduced separately for each agro-ecological zone in a Bayesian model to estimate the prevalence of T. parva infections and the sensitivity of the four diagnostic tests. All test specificities were set to 100%. The estimated T. parva prevalence was much higher (83-85%) than estimations based on single diagnostic tests. The estimated sensitivity of serological tests was relatively constant and ranged from 57 to 75% in the various areas. The sensitivity of PCR showed more pronounced variations, ranging from 66% in the low T. parva transmission (high land) zones compared to 24% in the highly vector infested (low land) zones. Calves and adult cattle (n=194) were also sampled in regularly and irregularly dipped herds in the low land region. The apparent T. parva prevalence detected by PCR was significantly higher in calves than in adult cattle and in herds regularly treated with acaricides, while no significant differences were found with IFAT. The conditional probability that a sample was positive at PCR while it was positive at IFAT was significantly lower in adults. The implication of these findings in the use of diagnostic assays for epidemiological studies is discussed. PMID- 18384964 TI - Role of the vomeronasal system in intersexual attraction in female mice. AB - Although it is generally accepted that rodents' sociosexual behavior relies mainly on chemosignals, the specific roles played by the vomeronasal and olfactory systems in detecting these signals are presently unclear. This work reports the results of three experiments aimed at clarifying the role of the vomeronasal system on gender recognition and intersexual attraction, by analyzing the effects of lesions of the accessory olfactory bulbs (AOB) in chemically naive female mice. The first experiment demonstrates that lesions of the AOB abolish the preference that females show for male-soiled bedding in tests in which the females can contact the bedding, thus having access to both volatile and involatile male chemosignals. The second experiment shows that airborne male derived chemosignals are not attractive to intact, chemically naive females but tend to be preferentially explored by females whose AOB has been lesioned. However, repeated exposure to male-soiled bedding has opposite effects in sham operated and AOB-lesioned female mice. Whereas after this experience sham operated females show an (acquired) attraction toward male airborne chemosignals, in AOB-lesioned females the same experience makes male-derived volatiles aversive. Finally, in the third experiment we have confirmed that our AOB lesioned females are able to detect urine-borne male odorants, as well as to discriminate them from the synthetic terpene geraniol. These findings strongly suggest that in mice, the involatile male sexual pheromone that is intrinsically attractive is detected by the vomeronasal system of the females. In addition, the repeated experience of females with male-soiled bedding would probably allow the association of this pheromone, acting as unconditioned stimulus, with olfactory stimuli (odorants) that therefore would become conditioned attractors to the females. PMID- 18384965 TI - Protective effects of ebselen, a seleno-organic antioxidant on neurodegeneration induced by hypoxia and reperfusion in stroke-prone spontaneously hypertensive rat. AB - Cerebral ischemia followed by oxygen reperfusion induced apoptosis in hippocampal neurons in the stroke-prone spontaneously hypertensive rat (SHRSP) but not in Wistar Kyoto rats (WKY). We investigated whether 2-phenyl-1,2-benzisoselenazol 3(2H)-one, also called PZ51 (ebselen), useful for treating ischemic damage or antihypertension in the brain, can protect against ischemic neuronal damage in SHRSP. In this study, we compared the effects of ebselen, carvedilol, 3-methyl-1 phenyl-2-pyrazolin-5-one (MCI-186) as well as vitamin E, added to cultures of neurons after reoxygenation (20% O(2)) following hypoxia (1% O(2)). SHRSP neurons died rapidly during reoxygenation following hypoxia but were rescued in large measure by 10 muM ebselen (neuronal death; 2.7+/-1.4%). In order of neuroprotective potency, the agents ranked as follows: ebselen>carvedilol>MCI 186>vitamin E. In vivo, strong neuroprotection by ebselen was observed in the hippocampal CA1 region of SHRSP (32.9+/-9.5 apoptotic neuron/1000 neurons, 30 mg/kg/day). Ebselen prevented apoptosis as confirmed by morphological observations in vivo. Its effect was associated with the expression of Bcl-2 and Bax. These findings suggest that ebselen has a marked inhibitory effect on neuronal damage during stroke. Ebselen may be effective in the prevention and/or treatment of neurodegenerative diseases associated with excessive apoptosis in patients with stroke. PMID- 18384963 TI - Neurochemistry of the afferents to the rat cochlear root nucleus: possible synaptic modulation of the acoustic startle. AB - Afferents to the primary startle circuit are essential for the elicitation and modulation of the acoustic startle reflex (ASR). In the rat, cochlear root neurons (CRNs) comprise the first component of the acoustic startle circuit and play a crucial role in mediating the ASR. Nevertheless, the neurochemical pattern of their afferents remains unclear. To determine the distribution of excitatory and inhibitory inputs, we used confocal microscopy to analyze the immunostaining for vesicular glutamate and GABA transporter proteins (VGLUT1 and VGAT) on retrogradely labeled CRNs. We also used reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry to detect and localize specific neurotransmitter receptor subunits in the cochlear root. Our results show differential distributions of VGLUT1- and VGAT-immunoreactive endings around cell bodies and dendrites. The RT-PCR data showed a positive band for several ionotropic glutamate receptor subunits, M1-M5 muscarinic receptor subtypes, the glycine receptor alpha1 subunit (GlyRalpha1), GABAA, GABAB, and subunits of alpha2 and beta-noradrenergic receptors. By immunohistochemistry, we confirmed that CRN cell bodies exhibit positive immunoreaction for the glutamate receptor (GluR) 3 and NR1 GluR subunits. Cell bodies and dendrites were also positive for M2 and M4, and GlyRalpha1. Other subunits, such as GluR1 and GluR4 of the AMPA GluRs, were observed in glial cells neighboring unlabeled CRN cell bodies. We further confirmed the existence of noradrenergic afferents onto CRNs from the locus coeruleus by combining tyrosine hydroxylase immunohistochemistry and tract tracing experiments. Our results provide valuable information toward understanding how CRNs might integrate excitatory and inhibitory inputs, and hence how they could elicit and modulate the ASR. PMID- 18384966 TI - Spectral processing deficits in belt auditory cortex following early postnatal lesions of somatosensory cortex. AB - Induced or genetically based cortical laminar malformations in somatosensory cortex have been associated with perceptual and acoustic processing deficits in mammals. Perinatal freeze-lesions of developing rat primary somatosensory (S1) cortex induce malformations resembling human microgyria. Induced microgyria located in parietal somatosensory cortex have been linked to reduced behavioral detection of rapid sound transitions and altered spectral processing in primary auditory cortex (A1). Here we asked whether belt auditory cortex function would be similarly altered in rats with S1 microgyria (MG+). Pure-tone acoustic response properties were assessed in A1 and ventral auditory (VAF) cortical fields with Fourier optical imaging and multi-unit recordings. Three changes in spectral response properties were observed in both A1 and VAF in MG+ rats: 1) multi-unit response magnitudes were reduced 2) optical and multi-unit frequency responses were more variable; 3) at high sound levels units responded to a broader range of pure-tone frequencies. Optical and multi-unit pure-tone response magnitudes were both reduced for low sound levels in VAF but not A1. Sound level "tuning" was reduced in VAF but not in A1. Finally, in VAF frequency tuning and spike rates near best frequency were both altered for mid- but not high-frequency recording sites. These data suggest that VAF belt auditory cortex is more vulnerable than A1 to early postnatal induction of microgyria in neighboring somatosensory cortex. PMID- 18384967 TI - Aged-related loss of temporal processing: altered responses to amplitude modulated tones in rat dorsal cochlear nucleus. AB - Loss of temporal processing is characteristic of age-related loss of speech understanding observed in the elderly. Inhibitory glycinergic circuits provide input onto dorsal cochlear nucleus (DCN) projection neurons which likely serve to modulate excitatory responses to time-varying complex acoustic signals. The present study sought to test the hypothesis that age-related loss of inhibition would compromise the ability of output neurons to encode sinusoidally amplitude modulated (SAM) tones. Extracellular recordings were obtained from young and aged FBN rat DCN putative fusiform cells. Stimuli were SAM tones at three modulation depths (100, 50, and 20%) at 30 dB hearing level with the carrier frequency set to the unit's characteristic frequency. Discharge rate and synchrony were calculated to describe SAM responses. There were significant age-related changes in the shape and peak vector strength [best modulation frequency (BMF)] of temporal modulation transfer functions (tMTFs), with no significant age-related changes in rate modulation transfer functions (rMTFs) at BMF. Young neurons exhibited band-pass tMTFs for most SAM conditions while aged fusiform cells exhibited significantly more low-pass or double-peaked tMTFs. There were significant differences in tMTFs between buildup, pauser-buildup, and wide chopper temporal response types. Young and aged wide-choppers displayed significantly lower vector strength values than the other two temporal DCN response types. Age-related decreases in the number of pauser-buildup response types and increases in wide-chopper types reported previously, could account, in part, for the observed loss of temporal coding of the aged fusiform cell. Age related changes in SAM coding were similar to changes observed with receptor blockade of glycinergic inhibition onto fusiform cells and consistent with previously observed age-related loss of endogenous glycine levels and changes in normal adult glycine receptor function. DCN changes in SAM coding could, in part, underpin temporal processing deficits observed in the elderly. PMID- 18384968 TI - The dimensions and structural attachments of tip links in mammalian cochlear hair cells and the effects of exposure to different levels of extracellular calcium. AB - The tip links between stereocilia of acousticolateral hair cells have been suggested to contain cadherin 23 (CDH23) comprising an upper branched portion that is bound to a lower portion composed of protocadherin 15 (PCDH15). The molecular conformation of CDH23, its binding to PCDH15, the tip links, and mechanoelectrical transduction have all been shown previously to be sensitive to exposure to low levels of calcium. The aim of this study was to compare the characteristics of tip links in guinea-pig cochlear hair cells with reported features of the CDH23-PCDH15 complex. Tip links were examined using field emission scanning electron microscopy and transmission electron microscopy in conventional preparations and after treatment with the detergent Triton-X-100 or varying calcium concentrations in the extracellular solution. The results showed that tip links have a twisted double-stranded appearance with a branched upper region. They survived demembranation of the stereocilia by detergent suggesting that they have transmembrane domains at both ends. Their lengths, when fixed in the presence of 2 mM extracellular calcium, were approximately 150 nm. With prior exposure to 1 mM calcium their lengths were approximately 164 nm. The lengths in 50 microM calcium are similar ( approximately 185 nm) to those reported for CDH23 PCDH15 complexes in 100 microM calcium ( approximately 180 nm). Exposure to 1 microM calcium caused loss of tip links and an increased distance between the residual attachment sites. The data indicate that extracellular calcium concentration affects tip-link length. One model compatible with the recently proposed tip-link structure is that the CDH23 double strand undergoes calcium dependent unfolding, changing the length of the links. The bundle may also tilt in the direction of the tallest row of stereocilia as the tip link lengthens and then is lost. Overall, our data are consistent with a tip link composed of complexes of CDH23 and PCDH15 but do not rule out other possibilities. PMID- 18384969 TI - Expression of the small heat shock protein family in the mouse CNS: differential anatomical and biochemical compartmentalization. AB - The small heat shock proteins (sHsps) are a family of molecular chaperones defined by an alpha-crystallin domain that is important for sHsps oligomerization and chaperone activity. sHsps perform many physiological functions including the maintenance of the cellular cytoskeleton, the regulation of protein aggregation and modulate cell survival in a number of cell types including glial and neuronal cells. Many of these functions have been implicated in disease processes in the CNS and indeed sHsps are considered targets for disease therapy. Despite this, there is no study that systematically and comparatively characterized sHsps expression in the CNS. In the present study we have analyzed the expression of this gene family in the mouse brain by reverse-transcriptase polymerase chain reaction (RT-PCR), in situ hybridization and Western blotting. Gene expression analysis of the 10 known members of mammalian sHsps confirms the presence of 5 sHsps in the CNS. A distinct white matter specific expression pattern for HspB5 and overlapping expression of HspB1 and HspB8 in the lateral and dorsal ventricles of the brain is observed. We confirm protein expression of HspB1, HspB5, HspB6 and HspB8 in the brain. Further subcellular fractionation of brain and synaptosomes details a distinct subcompartment-specific association and detergent solubility of sHsps. This biochemical signature is indicative of an association with synaptic and other neural specializations. This observation will help one understand the functional role played by sHsps during physiology and pathology in the CNS. PMID- 18384970 TI - Connexin31.1 deficiency in the mouse impairs object memory and modulates open field exploration, acetylcholine esterase levels in the striatum, and cAMP response element-binding protein levels in the striatum and piriform cortex. AB - Neuronal gap junctions in the brain, providing intercellular electrotonic signal transfer, have been implicated in physiological and behavioral correlates of learning and memory. In connexin31.1 (Cx31.1) knockout (KO) mice the coding region of the Cx31.1 gene was replaced by a LacZ reporter gene. We investigated the impact of Cx31.1 deficiency on open-field exploration, the behavioral response to an odor, non-selective attention, learning and memory performance, and the levels of memory-related proteins in the hippocampus, striatum and the piriform cortex. In terms of behavior, the deletion of the Cx31.1 coding DNA in the mouse led to increased exploratory behaviors in a novel environment, and impaired one-trial object recognition at all delays tested. Despite strong Cx31.1 expression in the peripheral and central olfactory system, Cx31.1 KO mice exhibited normal behavioral responses to an odor. We found increased levels of acetylcholine esterase (AChE) and cAMP response element-binding protein (CREB) in the striatum of Cx31.1 KO mice. In the piriform cortex the Cx31.1 KO mice had an increased heterogeneity of CREB expression among neurons. In conclusion, gap junctions featuring the Cx31.1 protein might be involved in open-field exploration as well as object memory and modulate levels of AChE and CREB in the striatum and piriform cortex. PMID- 18384971 TI - Effects of phase duration and pulse rate on loudness and pitch percepts in the first auditory midbrain implant patients: Comparison to cochlear implant and auditory brainstem implant results. AB - The auditory midbrain implant (AMI), which is designed for stimulation of the inferior colliculus (IC), is now in clinical trials. The AMI consists of a single shank array (20 contacts) and uses a stimulation strategy originally designed for cochlear implants since it is already approved for human use and we do not yet know how to optimally activate the auditory midbrain. The goal of this study was to investigate the effects of different pulse rates and phase durations on loudness and pitch percepts because these parameters are required to implement the AMI stimulation strategy. Although each patient was implanted into a different region (i.e. lateral lemniscus, central nucleus of IC, dorsal cortex of IC), they generally exhibited similar threshold versus phase duration, threshold versus pulse rate, and pitch versus pulse rate curves. In particular, stimulation with 100 mus/phase, 250 pulse per second (pps) pulse trains achieved an optimal balance among safety, energy, and current threshold requirements while avoiding rate pitch effects. However, we observed large differences across patients in loudness adaptation to continuous pulse stimulation over long time scales. One patient (implanted in dorsal cortex of IC) even experienced complete loudness decay and elevation of thresholds with daily stimulation. Comparing these results with those of cochlear implant and auditory brainstem implant patients, it appears that stimulation of higher order neurons exhibits less and even no loudness summation for higher rate stimuli and greater current leakage for longer phase durations than that of cochlear neurons. The fact that all midbrain regions we stimulated, which includes three distinctly different nuclei, exhibited similar loudness summation effects (i.e. none for pulse rates above 250 pps) suggests a possible shift in some coding properties that is affected more by which stage along the auditory pathway rather than the types of neurons are being stimulated. However, loudness adaptation occurs at multiple stages from the cochlea up to the midbrain. PMID- 18384972 TI - Non-plastic reorganization of frequency coding in the inferior colliculus of the rat following noise-induced hearing loss. AB - It is well established that restricted mechanical lesions of the cochlea result in reorganization of the tonotopic map in the auditory thalamus and cortex, but it is unclear whether acoustic trauma produces similar effects at earlier stages of the auditory pathways. To test whether the tonotopic map is reorganized after acoustic trauma at the midbrain level, i.e. the inferior colliculus (IC), we exposed rats to an acoustic trauma and let them survive for at least 5 weeks to ensure that we produced a permanent threshold shift. Experiments were carried out in urethane-anesthetized animals 35-296 days after the traumatic exposure. The acoustic lesions were assessed by measuring the compound action potential. We mapped the frequency organization of the IC using multiunit recordings. In addition, we recorded frequency response areas (FRAs) when a single unit was isolated (N=142). The results show that acoustic trauma produces a persistent reorganization of the tonotopic map and that the normal stepwise representation of sound frequency in the IC is profoundly disrupted. Although the reorganization in the IC is similar to that previously described in the cortex and thalamus in that the affected area appears to be invaded by the adjacent normal frequencies, changes in thresholds and FRAs in these regions are different from those in the forebrain. We conclude that most of the changes can be explained by the residual response hypothesis [Irvine DR, Rajan R, Smith S (2003) Effects of restricted cochlear lesions in adult cats on the frequency organization of the inferior colliculus. J Comp Neurol 467:354-374]. Plastic reorganization of frequency response areas and tonotopic organization does not seem to occur at the midbrain level following acoustic trauma in adult animals in a manner similar to that previously shown in the auditory cortex. Maintaining the stability of the neuronal circuitry for frequency coding in the IC may be important for the treatment of noise-induced hearing loss. PMID- 18384973 TI - Using an item response model to examine the nicotine dependence construct as characterized by the HONC and the mFTQ among adolescent smokers. AB - Despite efforts to operationalize the nicotine dependence syndrome among adolescents, little is known regarding the relative severity of dependence symptoms assessed by different measures. The current study utilized a nonparametric item response model to assess the unidimensionality of the nicotine dependence construct and relative severity of dependence symptoms characterized by the Hooked on Nicotine Checklist (HONC) and the modified Fagerstrom Questionnaire (mFTQ) in a sample of 109 adolescent smokers (58% female) participating in a prospective investigation of smoking self-change efforts. It was hypothesized that symptoms assessed by the HONC would be associated with lower levels of nicotine dependence severity than symptoms assessed with the mFTQ. Results indicated that HONC and mFTQ items could be linked to a single latent construct. Most HONC items captured variability at the lower range and mFTQ items made discriminations at the middle and higher end of the dependence severity continuum. Findings suggest the HONC and mFTQ may provide complementary information in assessing nicotine dependence levels in adolescent regular smokers and have implications for symptoms expression in youth. PMID- 18384974 TI - Relationship of perceived risks of smoking cessation to symptoms of withdrawal, craving, and depression during short-term smoking abstinence. AB - The current study examined the relationship between perceived risks of quitting smoking and the self-reported experience of risks (e.g., cravings) during short term abstinence. Participants (n=55) were daily smokers enrolled in a study of contingency management for smoking cessation with biochemically confirmed abstinence for 1 week. Participants were divided into groups of higher and lower perceived risk of quitting. There were no differences by risk group in demographics or baseline smoking, nicotine dependence, cravings, withdrawal, and depression. Although participants with higher levels of perceived risks reported a similar pattern of cravings and withdrawal symptoms during abstinence, they experienced higher levels of cravings, withdrawal symptoms, and depression than participants with lower risk beliefs. There were no differences in the relationship of risk to withdrawal symptoms by gender. Smokers with higher levels of perceived risk may find it more difficult to quit and remain abstinent due to higher levels of anticipated or experienced withdrawal symptoms and may benefit from targeted behavioral interventions with regard to risk perception and enhanced coping with withdrawal and other effects of smoking cessation. PMID- 18384975 TI - Workplace harassment, stress, and drinking behavior over time: gender differences in a national sample. AB - Research suggests that workplace harassment (WH) significantly predicts alcohol use and problem drinking behavior, but has generally failed to consider concurrent effects of other sources of stress. This two-wave study (n=1418) is the first to explore whether sexual harassment (SH) and generalized workplace harassment (GWH) predict increased drinking independently of the effects of job and life stress, and whether effects differ by gender, in a nationally representative sample. SH and GWH predicted increases in problem drinking one year later for men but not women, while life stress was associated with increased problem drinking for women but not men. This study confirms the importance of examining the associations between different types of stressors and drinking related outcomes in gendered contexts. PMID- 18384976 TI - Motivation of trauma patients to stop smoking after admission to the emergency department. AB - Every smoker should be offered smoking cessation treatment when they present for clinical care. The Readiness to Change-Smokers (RTC-S) questionnaire and the Heidelberg Smoking History (HSH) are brief questionnaires that divide patients into three stages. The purpose of this study was to prospectively compare the performance of each questionnaire at identifying patients who will successfully quit smoking within one year of Emergency Department (ED) discharge. Out of 1292 injured ED patients nearly half (n = 599, 46.4%) were identified as current smokers. Both questionnaires were given to all 599 subjects, and used to divide patients into three stages. At 12-months postdischarge 306 patients (51.1%) were contacted to determine smoking status. Patients were similarly classified by both tests in only 36% of cases. Concordance between tests was poor (kappa = 0.33). The RTC-S classified fewer patients as ready to quit (A = 13% vs. 22.2%). At 12 month follow-up, 55 patients (17.9%) had stopped smoking. The HSH was more successful to predict quitters. Multivariate logistic regression with respect to smoking cessation resulted in significant impact of HSH (p = 0.024). PMID- 18384977 TI - Comparing the generalized matching law and contingency discriminability model as accounts of concurrent schedule performance using residual meta-analysis. AB - To compare the generalized matching law (Baum, W.M., 1974b. On two types of deviation from the matching law: bias and undermatching. J. Exp. Anal. Behav. 22, 231-242) and contingency discriminability model (Davison, M., Jenkins, P.E., 1985. Stimulus discriminability, contingency discriminability, and schedule performance. Anim. Learn. Behav. 13, 77-84) as accounts of concurrent schedule performance, we conducted a residual meta-analysis of response- and time allocation data from 20 studies (n's=886 and 774, respectively). Both models were fitted to the individual-subject data from each study, and residuals were obtained. Polynomial regressions were then performed on the pooled residuals to determine whether systematic trends were present as a function of predicted values. For the contingency discriminability model, the cubic coefficients were positive and statistically significant for both response- and time-allocation data. By contrast, no statistically significant systematic trend was obtained in the residuals for the generalized matching law. These results suggest that the relationship between log response allocation and log reinforcer allocation does not deviate significantly from linearity over an approximate range of +1.25 to 1.25 log units, consistent with the generalized matching law. Although qualitative criteria are also important in comparing models of behavioral phenomena, residual meta-analysis provides a powerful quantitative methodology for model selection and should prove useful in future research. PMID- 18384978 TI - Association of candidate genes with antisocial drug dependence in adolescents. AB - The Colorado Center For Antisocial Drug Dependence (CADD) is using several research designs and strategies in its study of the genetic basis for antisocial drug dependence in adolescents. This study reports single nucleotide polymorphism (SNP) association results from a targeted gene assay (SNP chip) of 231 primarily Caucasian male probands in treatment with antisocial drug dependence and a matched set of community controls. The SNP chip was designed to assay 1500 SNPs distributed across 50 candidate genes that have had associations with substance use disorders and conduct disorder. There was an average gene-wide inter-SNP interval of 3000 base pairs. After eliminating SNPs with poor signals and low minor allele frequencies, 60 nominally significant associations were found among the remaining 1073 SNPs in 18 of 49 candidate genes. Although none of the SNPs achieved genome-wide association significance levels (defined as p<.000001), two genes probed with multiple SNPs (OPRM1 and CHRNA2) emerged as plausible candidates for a role in antisocial drug dependence after gene-based permutation tests. The custom-designed SNP chip served as an effective and flexible platform for rapid interrogation of a large number of plausible candidate genes. PMID- 18384980 TI - Disposition of styrene-acrylonitrile (SAN) trimer in female rats: single dose intravenous and gavage studies. AB - Styrene-acrylonitrile trimer (SAN Trimer), a mixture of six isomers (four isomers of 4-cyano-1,2,3,4-tetrahydro-alpha-methyl-1-naphthaleneacetonitrile [THAN] and two isomers of 4-cyano-1,2,3,4-tetrahydro-1-naphthaleneproprionitrile [THNP]), is a by-product of a specific production process of styrene-acrylonitrile polymer. Disposition studies in female rats were conducted to evaluate the pharmacokinetic behavior of [3H]SAN Trimer following a single intravenous administration (26 mg/kg) to nonpregnant rats; a single gavage administration (nominal doses of 25 mg/kg, 75 mg/kg, or 200 mg/kg in corn oil) to nonpregnant rats; and a single gavage administration (nominal dose of 200 mg/kg in corn oil) to pregnant and lactating rats. SAN Trimer was rapidly eliminated from blood (T1/2 approximately 1h) following a single intravenous dose and following single oral doses (T1/2 approximately 3-4h). SAN Trimer was also rapidly excreted in the urine and feces following single oral doses, while total radioactivity was cleared more slowly. In pregnant rats, the concentrations of both radioactivity and SAN Trimer 2h after dosing were highest in the blood, followed by the placenta, with the lowest levels in the fetus. In lactating rats, the concentrations of both radioactivity and SAN Trimer were higher in milk than in maternal blood. Total radioactivity and SAN Trimer blood concentrations in nonpregnant, pregnant, and lactating rats were both higher in lactating rats compared to nonpregnant and pregnant rats. PMID- 18384979 TI - Reward-related decision-making deficits and elevated impulsivity among MDMA and other drug users. AB - BACKGROUND: The recreational drug, 3,4-methylenedioxymethamphetamine (MDMA; 'Ecstasy'), is a synthetic amphetamine derivative and a serotonin neurotoxin. MDMA use is associated with cognitive dysfunction and impulsivity, but since polydrug abuse is common among users it is difficult to attribute these problems specifically to MDMA. Moreover, few studies have examined reward-related cognitive processes. Our aim was to examine reward-related decision-making and impulsivity among MDMA users while controlling for polydrug use via appropriate comparison groups. METHODS: We examined decision-making [Iowa Gambling Task, IGT; Bechara, A., Damasio, A.R., Damasio, H., Anderson, S.W., 1994. Insensitivity to future consequences following damage to human prefrontal cortex. Cognition 50, 7 15], self-reported impulsivity (Multidimensional Personality Questionnaire-Brief Form [constraint subscale]; Barratt Impulsiveness Scale; Zuckerman Sensation Seeking Scale), and drug use among 22 abstinent MDMA users, 30 other drug users, and 29 healthy non-drug controls. RESULTS: MDMA and other drug users showed comparable patterns of decision-making and impulsivity. However, both drug groups demonstrated poorer IGT performance and elevated self-reported impulsivity relative to controls. Poorer decision-making was related to heavier drug use in the past year, heavier weekly alcohol use, and meeting lifetime substance use disorder (SUD) criteria for more drug classes. Elevated impulsivity was associated with heavier drug use, heavier weekly alcohol use, more lifetime SUDs, and higher self-reported depression levels. CONCLUSIONS: These findings contradict the idea that MDMA is specifically associated with deficient decision making. Drug users, in general, may be at risk for decision-making deficits and elevated impulsivity. Such behaviors may represent trait factors that lead to the initiation of drug and alcohol use, and/or they may represent behavior patterns that are exacerbated by extensive use. PMID- 18384981 TI - The menopausal hot flush--anything new? AB - Although the hot flush is generally recognised by women and the medical profession as the most characteristic and often a very distressing symptom of the climacteric, it remains an enigma. The physiological changes associated with the hot flush are different from any other flushing condition, with an increased peripheral blood flow, increased heart rate and in particular a decrease in galvanic skin resistance, which is unique to the flush. Flushing occurs as a result of disturbance of the temperature regulating mechanism situated in the hypothalamus, and probably a reduction in the thermoneutral zone, within which fluctuations of basal body temperature do not provoke compensatory vascular responses. Many factors have been implicated, including hormone releasing factors, gonadotrophins and neurohumorals. However, the role of oestrogen is critical and the clinical value of oestrogen therapy is well established and has been confirmed by a Cochrane review. Nevertheless, the precise mechanism by which reduced circulating levels of oestrogen are involved in causing the flush has not yet been established. Priming with oestrogen seems to be an essential pre requisite for flushing, as young women with ovarian dysgenesis and very low circulating levels of oestrogen never have hot flushes unless they are given oestrogen replacement therapy, which is later discontinued. Oestrogen antagonist activity by selective oestrogen receptor modulators such as tamoxifen and raloxifene can also cause flushing. A link with gonadotrophins is demonstrated by a temporal association of flushes with the pulsatile release of luteinising hormone (LH). However, if LH pulses are eliminated by GnRH analogue, the frequency of flushing is not altered, which confirms that LH is merely associated with the flush rather than being causative. It is probable that the flush is initiated by a supra-pituitary mechanism which is influenced by the hypothalamic factors responsible for pulsatile LH release. A variety of chemical pathways have been proposed involving serotonin, noradrenalin and dopamine. Trials of drugs that selectively inhibit the re-uptake of serotonin and noradrenalin have shown some beneficial effects, as also has gabapentin, but often the results have been disappointing, and certainly less than the response seen with oestrogen or tibolone. The prevalence of hot flushes varies considerably around the world and is less in the Far East than in the west. Differences in diet and in particular the intake of phytoestrogens has been implicated and many studies have tried to establish whether dietary supplementation with phytoestrogens might be a suitable alternative to conventional hormone replacement therapy (HRT). So far, the results are disappointing. Other lifestyle measures such as avoiding alcohol, caffeine and spicy foods, keeping the core body temperature cool, paced respiration, taking exercise and even acupuncture may help. Hot flushes remain a major cause of reduced quality of life in a large proportion of menopausal women, but perhaps because they are not fatal and are usually self-limiting, there has been rather limited research or clinical interest. However, for the increasing number of women being treated with tamoxifen for breast cancer, and for whom oestrogen will usually be contra-indicated or unsuitable, there is an urgent need to identify the underlying mechanism so that appropriate, specific and safe non oestrogen therapy can be offered to improve their quality of life. PMID- 18384982 TI - Cationic lipid-coated magnetic nanoparticles associated with transferrin for gene delivery. AB - Cationic lipid-coated magnetic nanoparticles (MPs) associated with transferrin were evaluated as gene transfer vectors in the presence of a static magnetic field. MPs were prepared by chemical precipitation and were surface-coated with cationic lipids, composed of DDAB/soy PC (60:40 mole/mole). These cationic MPs were then combined with polyethylenimine (PEI) condensed plasmid DNA, followed by transferrin. The resulting magnetic electrostatic complexes retained relatively compact particle size and showed complete DNA condensation. Their transfection activity in the presence of a static magnetic field was evaluated by luciferase and green fluorescent protein (GFP) reporter genes. The magnetic complexes exhibited up to 300-fold higher transfection activity compared to commonly used cationic liposomes or cationic polymer complexes, based on luciferase assay. The enhancement in transfection activity was maximized when the cells were exposed to the vectors for a relatively short period of time (15 min), or were treated in media containing 10% serum. Incorporation of transferrin further improved transfection efficiency of the cationic MPs. However, when cells were incubated for 4h in serum-free media, magnetic and non-magnetic vectors showed similar transfection efficiencies. In conclusion, transferrin-associated cationic MPs are excellent gene transfer vectors that can mediate very rapid and efficient gene transfer in vitro in the presence of a magnetic field. PMID- 18384983 TI - Chitosan microspheres for encapsulation of alpha-lipoic acid. AB - Encapsulation of alpha-lipoic acid (LA) was carried out using chitosan as an encapsulant matrix. Placebo and LA-loaded chitosan microspheres were prepared by a spray-drying process. Scanning electron microscopy (SEM) studies confirmed the spherical particle geometry and the smooth surface morphology of LA-loaded particles. The particle size distribution (PSD) analysis of the placebo and LA loaded microspheres has shown that 50% of the microspheres were less than 3.53 and 7.89 microm, respectively. The structural interactions of the chitosan matrix with the encapsulated LA were studied by Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) which revealed structural interactions of lipoic acid with the encapsulant matrix. The antioxidant activity of encapsulated lipoic acid was studied using the free-radical scavenging assay. This study demonstrated significant retention of antioxidant activity of lipoic acid (75%) after encapsulation in the chitosan matrix. Encapsulation efficiency of lipoic acid obtained in this study was 55.2% when ethanol and acetic acid (1:1 v/v) was used as incubation/extraction medium. PMID- 18384984 TI - Stability of proteins encapsulated in injectable and biodegradable poly(lactide co-glycolide)-glucose millicylinders. AB - PURPOSE: To characterize protein stability in poly(lactide-co-glycolide) 50/50 glucose star (PLGA-Glu) injectable millicylinders and to compare results with linear PLGA 50/50. METHODS: Bovine serum albumin (BSA), a model protein, was encapsulated in PLGA-Glu and linear PLGA millicylinders by solvent-extrusion and incubated under physiological conditions. Important system properties were characterized, including: polymer molecular weight distribution, soluble acidic residues, polymer morphology, polymer water uptake, microclimate pH, protein content and release, and protein aggregation. The polymer microclimate late in the release incubation was simulated and protein recovery was analyzed by UV280, size exclusion chromatography, amino acid analysis, and a modified Bradford assay. RESULTS: PLGA-Glu contained higher levels of low molecular weight oligomers, more rapidly biodegraded, and exhibited a lower microclimate pH than the linear 50/50 PLGA, which is the most acidic type in the PLGA family. BSA, when encapsulated in PLGA-Glu millicylinders, underwent extensive noncovalent insoluble aggregation over 2 weeks in vitro release, which was almost completely inhibited upon co-encapsulation of Mg(OH)2. However, by 5 weeks release for base containing formulations, although insoluble aggregation was still suppressed, the soluble fraction of protein in the polymer was unrecoverable by the modified Bradford assay. Polymer microclimate simulations with extensive protein analysis strongly suggested that the low recovery was mostly caused by base-catalyzed hydrolysis of the oligomeric fraction of BSA. CONCLUSIONS: In PLGA-Glu, the acidic microclimate was similarly responsible for insoluble aggregation of encapsulated BSA. BSA aggregation was inhibited in millicylinders by co incorporation into the polymer an insoluble base, but over a shorter release interval than linear PLGA likely because of a more acidic microclimate in the star polymer. PMID- 18384985 TI - Noise-induced hearing loss in mice treated with antiretroviral drugs. AB - The results reported here for CBA/CaJ mice describe the effects of regular dosing with a common antiretroviral drug combination on outer hair cell (OHC) function using measures of 2f1-f2 distortion product otoacoustic emissions (DPOAEs) and auditory brainstem responses (ABRs). Specifically, experimental mice were treated daily over a 3-mo period with the nucleoside reverse transcriptase inhibitors (NRTIs), zidovudine (ZDV) and lamivudine (3TC), dissolved in their drinking water, while their control counterparts received untreated water. DPOAE levels and ABR detection thresholds prior to and after 12 wk of NRTI treatment did not differ between experimental and control groups. To assess whether NRTI treatment potentiates the adverse effects of noise over-exposure on OHC function, both experimental and control mice were exposed 1 wk later, while still on the drug regimen, to a 10-kHz octave-band noise (OBN) at 105-dB SPL for 1h. A major outcome of the sound over-exposure episode was that the NRTI-pretreated mice showed significantly greater permanent OBN-induced reductions in DPOAE levels at 2 wk postexposure than were observed for the untreated control animals. These findings support the notion that a synergistic relationship exists between certain NRTIs and intense sounds in that such preexposure drug treatments produced greater noise-induced decreases in DPOAE activity than did noise exposure alone. This drug/noise interaction is consistent with the known harmful effects of NRTIs on cellular mitochondrial activity. PMID- 18384986 TI - Evolvulus alsinoides (Convolvulaceae): an American herb in the Old World. AB - People in the Indian region often apply shankhapushpi and vishnukranti, two Sanskrit-based common names, to Evolvulus alsinoides. These are pre-European names that are applied to a medicinal American species transported into the area. The period of introduction is uncertain, but probably took place in the 1500s or 1600s. Examination of relationships of Evolvulus alsinoides, geographic distribution, its names in Asia, medical uses, and chemical and laboratory analysis indicates that the alien plant was adopted, given an ancient Indian name, and incorporated into some Old World pharmacopoeias. The herb apparently was included in medicines because it not only reminded people of certain aspects of their gods and goddesses, but also because the chemicals it contained were useful against some maladies. PMID- 18384987 TI - An inventory of the ethnoveterinary practices for reproductive disorders in cattle and buffaloes, Sargodha district of Pakistan. AB - AIM OF THE STUDY: This study was conducted to document the ethnoveterinary practices (EVPs) used for the treatment and/or management of reproductive disorders of cattle and buffaloes. MATERIALS AND METHODS: The information was collected using rapid and participatory rural appraisal techniques through interviews and focused group discussions with 217 traditional veterinary healers (TVHs) over a period of 16 months. RESULTS: The use of 66 plant species was documented for the treatment of reproductive disorders in the two dairy species. Most frequently reported (>or=20 times) plants represented Apiaceae, Poaceae, Pedaliaceae, Linaceae, Arecaceae, Brassicaceae, Malvaceae, Zingiberaceae, Rosaceae, Cannabaceae, Fabaceae, Moraceae, Rubiaceae and Mimosaceae families. All the documented plant species were indigenous to the study area. Genital prolapse, retention of fetal membranes, silent estrous/delayed puberty, anestrous, dystokia associated with incomplete cervical dilatation and retention of lochial discharge were the frequently reported reproductive disorders. Some of the plants were reported to be used in more than one condition. Materials other than plants used for the treatment of these disorders included camel milk, butter and butter fat, minerals, eggs, organic matters, chemical substances and meat preparations. These materials were used either alone or as adjunctive therapy. CONCLUSION: Richness of EVPs in the study area and extensive variation in the doses, methods of preparation, indications, and claims regarding efficacy of plants for various disorders merit controlled studies for their validation. PMID- 18384988 TI - Anti-mycobacterial activity of five plant species used as traditional medicines in the Western Cape Province (South Africa). AB - ETHNOPHARMACOLOGICAL RELEVANCE: Five plants used in traditional medicine in the Western Cape Province of South Africa, have been investigated for anti mycobacterial activity: Olea capensis, Tulbaghia alliacea, Dittrichia graveolens, Leysera gnaphalodes and Buddleja saligna. AIM OF THE STUDY: The aim was to assess antimycobacterial activity in plants used in treatment of symptoms of TB, and through activity-guided fractionation of extracts to isolate compounds or mixtures with potential as anti-TB drug leads. MATERIALS AND METHODS: Extracts and derived fractions were assayed against strains of Escherichia coli, Staphylococcus aureus, and Mycobacterium aurum A+. Isolated pure compounds were further tested against Mycobacterium species M. avium ATCC 25291, M. scrofulaceum ATCC 19981, M. microti ATCC 19422 and Mtb H37Rv, and for cytotoxicity against Chinese hamster ovarian cells. RESULTS: Extracts of B. saligna and L. gnaphaloides exhibited significant anti-mycobacterial activity, primarily associated with the presence of non-cytotoxic triterpenoids oleanolic acid in B. saligna and both oleanolic and ursolic acids in L. gnaphaloides. CONCLUSIONS: Anti-mycobacterial activity of extracts of selected plants is consistent with their traditional use. The identification of oleanolic and ursolic acids in these plants, and verification of their activity, underlines the potential for exploring structure-activity relationships of derivatives of these ubiquitous triterpenoids. PMID- 18384989 TI - Anti-inflammatory and analgesic activities of ethanolic extract and two limonoids from Melia toosendan fruit. AB - AIM OF STUDY: The fruit of Melia toosendan Sieb. et Zucc. (MTF) is a traditional Chinese herbal medicine in the treatment of stomachache and many acute or chronic inflammations, as well as ascariasis. This paper aimed to investigate the anti inflammatory and analgesic activities of the MTF extract and two main limonoid type triterpenoids isolated from MTF. MATERIALS AND METHODS: The ethanolic extract of MTF and two limonoids, isotoosendanin (1) and 1-O-tigloyl-1-O debenzoylohchinal (2) were evaluated for their anti-inflammatory and analgesic activities. Acetic acid-induced vascular permeability and lambda-carrageenan induced hind paw edema tests in mice were used to investigate anti-inflammatory activity; and acetic acid-induced writhing and hot-plate tests in mice were used to determine analgesic effect. RESULTS: Both the ethanolic extract and two limonoids displayed significant anti-inflammatory effects. Although the ethanolic extract showed remarkable analgesic effects in both writhing and hot-plate tests, the two limonoids had analgesic effects just in writhing test. CONCLUSION: The results suggested that the ethanolic extract of MTF had obvious anti-inflammatory and analgesic activities, and the two limonoids were the active constituents contributing to the anti-inflammatory and analgesic effects of MTF. PMID- 18384990 TI - Studies on the toxicity of gambogic acid in rats. AB - AIM: To study the chronic toxicity of gambogic acid (GA), the major active ingredient of gamboges, a brownish to orange resin extracted from the Garcinia hanburyi (family Guttiferae) in Southeast Asia, using Sprague-Dawley rat as an animal model and provide further theoretical support for clinical applications of this promising natural anticancer agent. METHODS: GA was administered orally at dosages of 120, 60 and 30 mg/kg once every other day for a total of 13 weeks. Then we carried out the chronic toxicity studies including general body parameters, hematological, serum biochemistry, histopathological, and viscera examination. RESULTS: The results from the studies demonstrated that rats treated with high dose (120 mg/kg) of GA for a long time can lead to the damage on the kidney and liver. An innocuous dose was established to be 60 mg/kg after administration to rats for a total of 13 weeks at a frequency of one administration every other day. This dose was approximately 18.0 (body weight) or 9.6 (body surface area) times higher then that of the dose (200mg/60 kg, every other day) used for human trials. CONCLUSIONS: The studies demonstrated that the toxicity targets in the rats were the kidney and liver. These results provide further theoretical support for clinical applications of this promising natural anticancer agent. PMID- 18384991 TI - A case of Churg-Strauss syndrome with refractory otitis media. AB - OBJECTIVE: Churg-Strauss syndrome (CSS) is known as autoimmune vasculitis with peripheral eosinophilia after bronchial asthma and rarely has otological findings. We present a case of CSS with refractory otitis media and discuss the relationship between otological symptoms of CSS and eosinophilic otitis media. CASE REPORT: A 60-year-old woman had suffered from recurrent sinusitis for 8 years, and also otitis media with effusion for 4 months. Eruption with peripheral eosinophilia was found in the lower legs; therefore, she was diagnosed with CSS. She was treated with systemic administration of prednisolone, intratympanic injection of betamethasone, and betamethasone nasal spray; thereafter, eosinophilia, otitis media and sinusitis rapidly improved. CONCLUSION: The features of eosinophilic otitis media are similar to the otological symptoms of CSS. It should be considered whether patients with eosinophilic otitis media have early phase CSS. PMID- 18384992 TI - Spinal muscular atrophy type I mimicking critical illness neuropathy in a paediatric intensive care neonate: electrophysiological features. AB - We report the case of a neonate with spinal muscular atrophy type I (SMA type I or Werdnig-Hoffman disease) who was initially misdiagnosis as having critical illness neuropathy. Electromyography (EMG) showed a moderate loss of voluntary and motor unit potentials of both neurogenic and myopathic appearance. Nerve conduction studies revealed the presence of a severe sensory-motor axonal neuropathy. Finally, a biopsy of quadriceps was compatible with the diagnosis of SMA type I. A genetic study confirmed the existence of a homozygous absence of exons 7 and 8 of the telomeric supervival motoneuron gene (SMN1 gene). PMID- 18384993 TI - [Pulmonary artery intimal sarcoma]. AB - Pulmonary artery sarcoma is a rare tumor. We present a case of intimal sarcoma arising from right pulmonary artery and left lower pulmonary vein observed in a 44-year-old man with a non-productive cough. Computed tomographic scans and magnetic resonance imaging showing filling defect enhancement contributed early, suggesting the diagnosis of primary vascular tumor, hypothesis confirmed by pathologist findings. PMID- 18384994 TI - [Percutaneous retrieval of intravascular foreign bodies]. AB - Percutaneous extraction of intravascular foreign bodies is becoming an increasingly common technique. It is an effective and low aggressive way to avoid surgery. We report four new cases of percutaneous retrieval. PMID- 18384995 TI - MDCT angiography for detection of pulmonary emboli: comparison between equi iodine doses of iomeprol 400 mgI/mL and iodixanol 320 mgI/mL. AB - OBJECTIVES: To compare iomeprol 400 and iodixanol 320 in pulmonary artery MDCTA in subjects with suspected pulmonary embolism. METHODS: Eighty randomized subjects received equi-iodine intravenous doses (48 g) of iomeprol 400 (n=40) or iodixanol 320 (n=40), via power injector at 4 mL/s. Four-row (35 subjects) and 64 row (45 subjects) scanners were used. Lumen attenuation was determined on-site and by two off-site blinded readers in the main, lobar, segmental and subsegmental pulmonary arteries. Statistical comparison between groups was performed for demographics and lumen attenuation. RESULTS: There were no between group differences (p>0.05) in demographics. Pulmonary artery attenuation was significantly (p60 medical centers in North America, Latin America, and Europe). All isolates (40 675) were susceptibility tested using reference broth microdilution methods. Ceftobiprole inhibited 100% and >99% of tested S. aureus and coagulase-negative staphylococci at < or =4 and < or =8 microg/mL, respectively, although MIC90 values for oxacillin-resistant strains were 4-fold and 8-fold higher than oxacillin-susceptible isolates for the 2 groups. Ceftobiprole was also broadly active against Streptococcus pneumoniae, beta-hemolytic and viridans group streptococci, inhibiting >98% of isolates at < or =0.5 microg/mL. Although ceftobiprole was generally inactive against Enterococcus faecium, the majority of Enterococcus faecalis strains (95.7%) were inhibited at < or =4 microg/mL. This agent was similar in potency to the "3rd- and 4th-generation" cephems (MIC50 values, < or =0.06 microg/mL) for all tested Enterobacteriaceae. Although cefepime provided enhanced coverage against Klebsiella spp. (86.5% at < or =8 microg/mL versus 76.9-81.7% for ceftobiprole and ceftazidime), ceftobiprole and cefepime were superior to ceftazidime against Enterobacter spp. and Citrobacter spp. Against Pseudomonas aeruginosa, ceftobiprole was equal in potency to ceftazidime (MIC50, 2 microg/mL) and 2-fold more potent than cefepime. None of these agents inhibited >45% of Acinetobacter spp. at 8 mug/mL. Ceftobiprole is a new anti-MRSA beta-lactam with recognized activity against the most commonly occurring Enterobacteriaceae and P. aeruginosa, similar to that of extended-spectrum cephems. These characteristics warrant continued evaluation of the agent as empiric therapy for cSSSIs, and in pneumonia, especially in those institutions/regions where MRSA and P. aeruginosa may be prevalent. PMID- 18385001 TI - Tactical emergency medical support: physician involvement and injury patterns in tactical teams. AB - Medical support provided by physicians in police tactical teams has been firmly embraced by the medical community. Our study revisited the 1995 study inquiring into injury patterns in police tactical teams. A national survey was completed by 209 members of tactical teams throughout the country over a 6-week period. An electronic survey was submitted to the National Tactical Officers Association, the International Tactical Emergency Medical Support Association, and state tactical associations. Teams reporting physician utilization were 47% of the whole (69% were presentG, CYP3A5*3 G>A, GSTP1 313 A>G, GSTM1 deletion, GSTT1 deletion, MDR1 exon 21 G>T/A, MDR1 exon 26 C>T, MTHFR 677 C>T, MTHFR 1298 A>C, NR3C1 1088 A>G, RFC 80 G>A, TPMT 238 G>C, TPMT 460 G>A, TPMT 719 A>G, VDR intron 8 G>A, VDR FokI T>C) that have been implicated in the pharmacogenetics of ALL therapy were analyzed by TotalPlex amplification and SNP genotyping. We successfully amplified specific gene fragments using 16 pairs of primers in one PCR reaction tube with minimal spurious amplification products using TotalPlex amplification coupled to a multiplexed bead array detection system. The genotypes of 16 loci from 34 different genomic DNA (gDNA) samples derived using the TotalPlex system were consistent with the results of several standard genotyping methods, including automatic sequencing, PCR restriction fragment length polymorphism (RFLP) analysis, PCR, and allele-specific PCR (AS PCR). Thus, the TotalPlex system represents a useful method of amplification that can improve the time, cost, and sample size required for high-throughput pharmacogenetic analysis of SNPs. PMID- 18385011 TI - An improved PCR method for gender identification of eagles. AB - Eagles are sexually monomorphic and therefore it is difficult to determine their gender, which is a crucial need for management purposes. In this study, we have developed an improved gender identification method by exploiting length differences between the Chromo-Helicase-DNA binding protein (CHD)-Z and CHD-W genes of Spilornis cheela hoya. By comparing DNA sequences for CHD-W and CHD-Z from 10 species of Falconiformes eagles we designed universal gender identification PCR primers that exploit differences in product size. Standard agarose gels were shown to easily distinguish between the 148-bp CHD-ZW and the 258-bp CHD-W PCR products. When used with 28 samples of S. cheela hoya, our improved universal primers provided a fast and precise gender identification assay. PMID- 18385012 TI - Outcome of comprehensive psycho-educational interventions for young children with autism. AB - This paper evaluates comprehensive psycho-educational research on early intervention for children with autism. Twenty-five outcome studies were identified. Twenty studies evaluated behavioral treatment, 3 studies evaluated TEACCH and 2 studies evaluated the Colorado Health Sciences Project. Outcome studies are graded according to their scientific value, and subsequently graded according to the magnitude of results documented in the studies. Based on the available evidence, treatment recommendations are made and practice parameters are suggested. PMID- 18385013 TI - Direct assessment of size and shape of noncircular vena contracta area in functional versus organic mitral regurgitation using real-time three-dimensional echocardiography. AB - BACKGROUND: Vena contracta width (VCW) as an estimate of effective regurgitant orifice area (EROA) is an accepted parameter of mitral regurgitation (MR) severity. However, uncertainty exists in cases in which VCW at the same time appears narrow in 4-chamber (4CH) view and broad in 2-chamber (2CH) view as common in functional MR with noncircular or slit-like regurgitant orifices. We therefore hypothesized that new real-time 3-dimensional color Doppler echocardiography (RT3DE) can be used for direct assessment of the size and shape of vena contracta area (VCA) in an en face view and to determine the potential error of conventional VCW measurement on estimation of EROA. METHODS: RT3DE was performed in 57 patients with relevant MR of different etiologies. Manual tracing of VCA in a cross-sectional plane through the vena contracta was compared with VCW in 4CH and 2CH views. As a comparative approach to VCA-3D, EROA was calculated using the hemispheric and hemielliptic proximal isovelocity surface (PISA) area method. RESULTS: Direct measurement of VCA-3D was feasible in all patients within 2.6 +/- 0.7 minutes. RT3DE revealed significant asymmetry of VCA in functional compared with organic MR (P < .001). Among all patients, VCW-4CH and VCW-2CH correlated only moderately to VCA-3D (r =.77; r =.80). Mean VCW correlated and agreed best with VCA-3D (r =.90). VCA-3D correlated and agreed well with EROA by hemielliptic PISA (r = .96, mean error: -0.09 +/- 0.14 cm(2)) compared with significant underestimation of hemispheric PISA in noncircular lesions. CONCLUSIONS: Direct assessment of VCA using RT3DE revealed significant asymmetry of VCA in functional MR compared with organic MR, resulting in poor estimation of EROA by single VCW measurements. PMID- 18385014 TI - Accelerated stress real-time myocardial contrast echocardiography for the detection of coronary artery disease: comparison with 99mTc single photon emission computed tomography. AB - OBJECTIVE: The aims of this prospective study were to compare the diagnostic value of accelerated vasodilator stress real-time myocardial contrast echocardiography (MCE) and single photon emission computed tomography (SPECT) against coronary angiography and to evaluate whether the addition of MCE perfusion data improves the diagnostic accuracy of stress echocardiography. METHODS: A total of 103 patients with suspected or known stable coronary artery disease (CAD) underwent SPECT and accelerated high-dose dipyridamole (0.84 mg/kg intravenously for 4 minutes) atropine (up to 1 mg intravenously) stress real-time qualitative MCE. The presence of CAD was detected by coronary angiography. RESULTS: CAD defined as >or= 70% stenosis was detected in 77% of patients, whereas 86% of patients had CAD defined as >or= 50% stenosis. In a territory-by territory analysis, the concordance between MCE and SPECT in detecting perfusion defects varied from 72.8% (kappa = 0.386) to 89.3% (kappa = 0.642). There were no significant differences between MCE and SPECT in sensitivity, specificity, and diagnostic accuracy for identifying patients with CAD. Combining MCE and wall motion abnormality analysis significantly improved the sensitivity of the test compared with wall motion abnormality analysis alone. CONCLUSIONS: Accelerated vasodilator stress real-time MCE yields a good concordance with SPECT in detection of perfusion defects and a similar diagnostic value for the detection of CAD. The addition of MCE perfusion data improves the diagnostic value of stress echocardiography. PMID- 18385015 TI - Left atrial function and work after surgical ventricular restoration in postmyocardial infarction heart failure. AB - INTRODUCTION: The purpose of the current study was to examine the effect of surgical ventricular restoration in left atrial (LA) performance and left ventricular (LV) diastolic function in patients with congestive heart failure after extensive anterior myocardial infarction. METHODS: Seventeen consecutive patients (age 63 +/- 9 years, 14 male, 4 with diabetes) were studied prospectively. All patients underwent surgical ventricular restoration and concomitant coronary artery bypass surgery. LA volumes were calculated, as were peak LA kinetic energy (LAKE) and LA ejection fraction (LAEF). LV diastolic filling patterns were also assessed. RESULTS: Follow-up period was 1 year. LA size and volumes did not change significantly during the follow-up period. Two distinct groups of patients were identified: group A, in which LAEF and peak LAKE increased, and group B, in which LAEF and peak LAKE decreased. In both groups, the LAEF and peak LAKE appeared to increase or decrease simultaneously and this became evident from the first month after the surgery. However, 1 year after the operation, diastolic filling pattern was significantly different between the two groups. LA function showed improvement only in patients who demonstrated an improvement in indices of diastolic filling pattern. CONCLUSIONS: Despite improvement in clinical status and indices of LV systolic function in all patients who underwent surgical ventricular restoration for treatment of congestive heart failure as a result of LV dilatation secondary to large myocardial infarction, improvement of LA function occurs in patients with improved LV diastolic function. PMID- 18385016 TI - CD4+ T cells from type 1 diabetic and healthy subjects exhibit different thresholds of activation to a naturally processed proinsulin epitope. AB - Recent studies suggest that insulin is a primary autoantigen for type 1 diabetes. Several studies have identified preproinsulin (PPI) 76-90 as an immunodominant CD4+ T cell epitope. We developed a class II tetramer reagent using a modified PPI peptide with a lysine to serine substitution at position 88 (PPI 78-90(88S)) that has high binding affinity to DRA1*0101/DRB1*0401 (DR0401). Using this tetramer, positive responses were observed from both DR0401 healthy and type 1 diabetic subjects when T cells were stimulated with the PPI 78-90(88S) peptide. Seventy percent of these T cells proliferated in response to both the wild type PPI 76-90 and PPI 78-90(88S) peptides. However, when T cells were stimulated with wild type peptide and assayed with DR0401/PPI 78-90(88S), positive responses were only detected in the diabetic group but not in healthy subjects. When highly purified CD4+CD25-CD45RA+ T cells were stimulated with PPI 78-90(88S) peptide in the absence of antigen-presenting cells, T cells from the diabetic group were able to respond to peptide stimulation, while T cells from healthy subjects were not. These data suggest that T cells from type 1 diabetic subjects have a lower threshold of activation in response to PPI peptide stimulation as compared to healthy subjects. PMID- 18385017 TI - Highlighting the role of Ras and Rap during Dictyostelium chemotaxis. AB - Chemotaxis, the directional movement towards a chemical compound, is an essential property of many cells and has been linked to the development and progression of many diseases. Eukaryotic chemotaxis is a complex process involving gradient sensing, cell polarity, remodelling of the cytoskeleton and signal relay. Recent studies in the model organism Dictyostelium discoideum have shown that chemotaxis does not depend on a single molecular mechanism, but rather depends on several interconnecting pathways. Surprisingly, small G-proteins appear to play essential roles in all these pathways. This review will summarize the role of small G proteins in Dictyostelium, particularly highlighting the function of the Ras subfamily in chemotaxis. PMID- 18385018 TI - Beyond the "omics": translating science into improved health. PMID- 18385019 TI - Premalignant lesions. PMID- 18385020 TI - Osteonecrosis of the jaw. PMID- 18385021 TI - Infective endocarditis. PMID- 18385022 TI - Porous structures boost integration of host tissue with titanium implants. PMID- 18385024 TI - Combined technologies benefit patients with head and neck cancer. PMID- 18385025 TI - The effects of orthodontic therapy on periodontal health: a systematic review of controlled evidence. AB - BACKGROUND: Orthodontic therapy has been suggested to lead to an improved periodontal status through mechanisms such as increased ease of plaque removal and reduced occlusal trauma. The objective of the authors' systematic review was to compare contemporary orthodontic treatment with no intervention, by means of evaluating periodontal outcomes measured after end of treatment. METHODS: The authors completed electronic searches in eight databases (1980-2006) and hand searches in six dental journals (1980-2006). They extracted data using standardized forms and calculated weighted mean differences. RESULTS: Weak evidence from one randomized study and 11 nonrandomized studies suggested that orthodontic therapy was associated with 0.03 millimeters of gingival recession (95 percent confidence interval [CI], 0.01-0.04), 0.13 mm of alveolar bone loss (95 percent CI, 0.07-0.20) and 0.23 mm of increased pocket depth (95 percent CI, 0.15-0.30) when compared with no treatment. The effects of orthodontic therapy on gingivitis and attachment loss were inconsistent across studies. CONCLUSIONS: This systematic review identified an absence of reliable evidence describing positive effects of orthodontic treatment on periodontal health. The existing evidence suggests that orthodontic therapy results in small detrimental effects to the periodontium. PMID- 18385026 TI - The myth of instant orthodontics: an ethical quandary. AB - BACKGROUND: There is a clinical trend of using porcelain veneer restorations (PVRs) for the correction of malaligned anterior teeth. Use of PVRs for this purpose raises clinical and ethical dilemmas. TYPES OF STUDIES REVIEWED: A literature review of four different topics (PVR preparation, enamel thickness of anterior teeth, dentinal bonding adhesive effectiveness and PVR long-term success) was conducted to determine the optimal preparation for a successful PVR. The amount of tooth malalignment that may be corrected with a PVR without adversely affecting its success was calculated. RESULTS: The optimal preparation for a successful PVR may have dentin exposed in the body of the preparation. However, most of the preparation must be in enamel, and all the margins must end in enamel. The strength of a dentin bond varies greatly owing to a multistep, technique-sensitive cementation process and is weaker than an enamel bond. It is not possible to correct atypical gingival esthetics (uneven gingival margins, uneven papillae, short papillae and bulbous gingivae) resulting from malaligned teeth through use of PVRs. CONCLUSIONS: and CLINICAL IMPLICATIONS: Aligning a healthy tooth with a PVR is not a conservative procedure and more conservative treatment options (such as orthodontics, bleaching, direct bonding and enamelplasty) should be offered to the patient. In addition, the inability to restoratively improve gingival relationships with PVRs may result in achieving less-than-optimal esthetics. A clinician should present only treatment options that involve predictable, conservative restorations or that preserve healthy tooth structure. Aligning teeth with PVRs may create ethical dilemmas that can be resolved with the help of the American Dental Association Principles of Ethics and Code of Professional Conduct. PMID- 18385027 TI - Pulpalgia contributing to temporomandibular disorder-like pain: a literature review and case report. AB - BACKGROUND: Dentists need to be cognizant that temporomandibular disorder (TMD) like pain can be caused by a tooth pulpalgia. The author provides suggestive symptom characteristics and definitive diagnostic techniques. CASE DESCRIPTION: A patient had severe bilateral TMD-like pain, which increased when something cold touched a premolar and when the patient lay down, and which awakened her several times every night. The author identified the offending tooth and administered a ligamentary injection along the tooth, which eliminated her bilateral TMD-like pain. Occlusal adjustment of her tooth reduced her pain, and subsequent endodontic therapy eliminated her pain. To the author's knowledge, this is the first report of a pulpalgia in a posterior tooth causing bilateral TMD-like pain. CLINICAL IMPLICATIONS: Pulpalgia may cause symptoms that mimic TMD or may contribute to TMD signs and symptoms. When patients with TMD-like pain report feeling increased pain due to a cold stimulus' coming into contact with a tooth, practitioners should ensure that a pulpalgia is not contributing to their pain. PMID- 18385028 TI - Extirpation of the primary canine tooth follicles: a form of infant oral mutilation. AB - BACKGROUND: Ebinyo is a form of infant oral mutilation (IOM), widely practiced in rural areas of eastern Africa, in which traditional healers and other village elders extirpate the primary canine tooth follicles of infants by using crude, often unsterilized, instruments or utensils. Traditional folklore suggests that the underlying tooth follicles, thought to resemble worms, are the cause of high temperature, vomiting, loss of appetite and diarrhea in infants. In addition to the serious and potentially fatal immediate postsurgical complications, many of those who undergo this practice exhibit characteristic long-term adverse dentoalveolar effects. Children in these families also may be at greater risk of undergoing other mutilation rituals because of their cultural background. CASE DESCRIPTION: We report on the clinical and radiographic findings in five siblings who apparently were subjected to IOM as infants before immigrating to the United States. CLINICAL IMPLICATIONS: Although the practice of IOM is believed to be exceedingly rare in developed countries, it is important that dentists and allied dental personnel who treat refugees from areas of the world in which IOM is endemic be aware of the social factors behind this practice as well as be able to recognize its dental and psychological sequelae. PMID- 18385029 TI - Dietary supplements: clinical implications for dentistry. AB - BACKGROUND: Sales of dietary supplements in the United States continue to increase; patients who seek dental care more than likely are taking or contemplating taking dietary supplements. The author reviews the background of dietary supplements, highlights popular over-the-counter supplements available in the United States and addresses the issues of safety, efficacy and contraindications that merit consideration in clinical practice. METHODS: The author reviewed the literature regarding effectiveness of common dietary supplements and mechanisms of action with drug interactions. The author assessed properties of dietary supplements within the context of clinical implications for oral health care professionals. CONCLUSIONS: Dietary supplement use may be prevalent among patients seeking dental care. While some dietary supplements may be effective and safe, their biological activity may interfere with treatment or medications in oral health care. CLINICAL IMPLICATIONS: Oral health care professionals can ascertain patients' use of dietary supplements and evaluate them with respect to treatment. Resources for additional information are provided. PMID- 18385030 TI - An ultrastructural investigation of tissue-engineered pulp constructs implanted within endodontically treated teeth. AB - BACKGROUND: The authors conducted an ultrastructural scanning electron microscopic (SEM) investigation of tissue-engineered pulp constructs implanted within endodontically treated teeth. METHODS: Stem cells from human exfoliated deciduous teeth were seeded on a synthetic open-cell D,D-L,L-polylactic acid scaffold with or without the addition of bone morphogenic protein-2 and transforming growth factor beta1 to create pulp tissue constructs. The pulp constructs were implanted into 105 extracted human premolar teeth with a single root canal that had been cleaned and shaped by using rotary instrumentation in a crown-down manner to ISO size no. 35. RESULTS: An ultrastructural examination of the SEM micrographs at x2,000 magnification revealed cell adherence within all of the pulp constructs, with little difference between the scaffold types or with the addition of growth factors. CONCLUSIONS: These results support the proof-of concept that it is possible to implant tissue-engineered pulp constructs into teeth after cleaning and shaping. CLINICAL IMPLICATIONS: Future regenerative endodontic treatment may involve the cleaning and shaping of root canals followed by the implantation of vital dental pulp tissue constructs created in the laboratory. PMID- 18385031 TI - Colorado dental practitioners' attitudes and practices regarding tobacco-use prevention activities for 8- through 12-year-old patients. AB - BACKGROUND: Tobacco use is a leading risk factor for oral morbidities and mortalities such as oral cancers and periodontitis. This study characterizes the factors related to dentists and hygienists conducting tobacco-use prevention counseling with 8- through 12-year-old patients. METHODS: The study used a mailed survey of dentists (n = 434), orthodontists (n = 91) and hygienists (n = 160) practicing in Colorado to collect data on the practitioners' tobacco-use prevention counseling activity, demographic characteristics, barriers to counseling and attitudes toward tobacco use. The authors used multiple logistic regression to determine which variables were associated independently with a dental practitioner's counseling children. RESULTS: The response rate was 25.6 percent. Prevalence of tobacco-use prevention counseling for children was low (38 percent for dentists and 44 percent for hygienists). Among dentists, the perception of tobacco use in children as a problem, perceived effectiveness of counseling and perceived role of a dental practitioner in counseling children were associated positively with counseling. Lack of skills was associated negatively with counseling. Among hygienists, perceived role in counseling children was associated positively and lack of time was associated negatively with counseling. CONCLUSIONS: Few dental practitioners counsel 8- through 12-year old patients about tobacco use. Factors related to counseling appear to be amenable to education of dentists, in particular, regarding the importance of the problem, the effectiveness of counseling and skill development. CLINICAL IMPLICATIONS: Preventing tobacco use among children is integral to promoting patients' oral health. Training dentists through continuing education should increase the frequency of tobacco-use prevention counseling with children by both dentists and hygienists. PMID- 18385032 TI - Digital radiographic imaging: is the dental practice ready? AB - BACKGROUND: Digital radiographic imaging is slowly, but surely, replacing film based imaging. It has many advantages over traditional imaging, but the technology also has some drawbacks. The author presents an overview of the types of digital image receptors available, image enhancement software and the range of costs for the new technology. PRACTICE IMPLICATIONS. The expenses associated with converting to digital radiographic imaging are considerable. The purpose of this article is to provide the clinician with an overview of digital radiographic imaging technology so that he or she can be an informed consumer when evaluating the numerous digital systems in the marketplace. PMID- 18385033 TI - Selecting the best abutment for a single implant. PMID- 18385034 TI - Moving beyond customer service. PMID- 18385035 TI - What are the ethical issues to consider when deciding whether to treat my staff? PMID- 18385036 TI - Focus issue: mechanisms of gene regulation. AB - In this Focus Issue of Science Signaling, which complements the Science Special Issue on Gene Regulation (http://www.sciencemag.org/generegulation/), we examine the multilayered mechanisms that control how gene expression is turned on or off. Perspectives in this issue discuss how oncogenic Ras directs the silencing of tumor suppressor genes; how intrachromosomal looping brings enhancers and promoters together to stimulate gene expression; and how the abundance of a transcriptional coactivator affects nuclear receptor-mediated transcription. PMID- 18385037 TI - Silent assassin: oncogenic ras directs epigenetic inactivation of target genes. AB - Oncogenic transformation is associated with genetic changes and epigenetic alterations. A study now shows that oncogenic Ras uses a complex and elaborate epigenetic silencing program to specifically repress the expression of multiple unrelated cancer-suppressing genes through a common pathway. These results suggest that cancer-related epigenetic modifications may arise through a specific and instructive mechanism and that genetic changes and epigenetic alterations are intimately connected and contribute to tumorigenesis cooperatively. PMID- 18385038 TI - NFAT is well placed to direct both enhancer looping and domain-wide models of enhancer function. AB - Nuclear factor of activated T cells (NFAT) plays a central role in activating gene expression at the level of chromatin structure. A study now reveals that NFAT may also help to organize chromatin domains and enable enhancer-promoter communication. In activated T cells, inducible intrachromosomal looping occurs between the tumor necrosis factor-alpha (TNF-alpha) gene promoter and two NFAT dependent enhancers located at -9 kb and +3 kb. This topology places the TNF alpha gene and the adjacent lymphotoxin (LT) genes in separate loops, thereby allowing independent regulation of the TNF-alpha gene within a multigene locus. These findings build on other studies that indicate that NFAT is intimately associated with activities that disrupt nucleosomes within enhancers and mobilize nucleosomes across extensive chromatin domains linking enhancers and promoters. Taken together, these studies highlight NFAT as a factor that creates a chromatin environment that is permissive for both the recruitment and the clustering of factors that control transcription at promoters and enhancers. PMID- 18385039 TI - SRC-3 transcription-coupled activation, degradation, and the ubiquitin clock: is there enough coactivator to go around in cells? AB - Overexpression of nuclear receptor coactivators is a frequent event in breast cancer cells and is recognized as a key mechanism for these cells to maximize their oncogenic growth state. Steroid receptor coactivator-3 [(SRC-3), also known as amplified in breast cancer-1 or AIB1] is foremost among these overexpressed oncogenic coactivators, being overexpressed in most breast cancers. Because of its oncogenic potential, normal cells must carefully control its cellular concentration. We discuss how SRC-3 quantitatively influences estrogen-regulated gene transcription when it is at limiting concentrations in normal breast cells and at nonlimiting concentrations in breast cancer cells. Precise control of the cellular concentration of SRC-3 may thus serve as a mechanism for defining growth responses to estrogen receptors and other growth-promoting transcription factors. PMID- 18385040 TI - Epigenetic regulation identifies RASEF as a tumor-suppressor gene in uveal melanoma. AB - PURPOSE: Recently, a segregation study in families with uveal and cutaneous melanoma identified 9q21 as a potential locus harboring a tumor-suppressor gene (TSG). One of the genes in this area, RASEF, was then analyzed as a candidate TSG, but lack of point mutations and copy number changes could not confirm this. In this study, the RASEF gene was investigated for potential mutations and gene silencing by promoter methylation in uveal melanoma. METHODS: Eleven uveal melanoma cell lines and 35 primary uveal melanoma samples were screened for mutations in the RASEF gene by high-resolution melting-curve and digestion analysis. Expression of RASEF was determined by real-time RT-PCR in all cell lines and 16 primary uveal melanoma samples, and the methylation status of the promoter of the RASEF gene was analyzed and confirmed by direct sequencing. RESULTS: Mutation screening revealed a known polymorphism (R262C; C-->T) in exon 5 of the RASEF gene that displayed a normal frequency (54%). Of the primary uveal melanomas, 46% presented a heterozygous genotype, and 10 (91%) of 11 cell lines showed a homozygous genotype. Melting-curve analysis indicated loss of heterozygosity in at least two primary tumors. Low RASEF expression in the cell lines and primary tumors correlated with methylation of the RASEF promoter region. Homozygosity and methylation of the RASEF gene in primary tumors were associated with decreased survival (P = 0.019). CONCLUSIONS: Homozygosity, in combination with methylation, appears to be the mechanism targeting RASEF in uveal melanoma, and allelic imbalance at this locus supports a TSG role for RASEF. PMID- 18385042 TI - ApoB100,LDLR-/- mice exhibit reduced electroretinographic response and cholesteryl esters deposits in the retina. AB - PURPOSE: To evaluate the retinal phenotype of 7- and 14-month-old apoB100,LDLR-/- mice, a relevant animal model of lipid metabolism dysfunction. METHODS: Single flash electroretinograms were obtained from 7- and 14-month-old apoB100,LDLR-/- and control mice fed a standard diet under both scotopic and photopic conditions. Visual cycle retinoids were analyzed in eyes from dark-adapted mice. Retinal and choroidal vascularization was evaluated with scanning laser ophthalmoscopy. Fatty acids were analyzed in the retina. Esterified and free cholesterol was detected in eye cryosections. RESULTS: Scotopic and photopic b-wave amplitudes were significantly reduced in apoB100,LDLR-/- mice compared with control mice at 7 and 14 months of age (between -25% and -35% in 7-month-old animals and between -50% and -60% in 14-month-old animals at 25 cds/m2). Esterified cholesterol was found to accumulate at the basement of the retinal pigment epithelium in apoB100,LDLR-/ mouse eyes. On the contrary, no significant changes in the retinal profile of fatty acids and visual retinoids were observed in apoB100,LDLR-/- mice compared with control animals. CONCLUSIONS: The exclusive expression of apoB100 in LDL receptor-null mouse altered the ERG profile, without modifying the visual cycle of retinoids and led to cholesterol deposition in the retina. These findings clearly suggest the role of cholesterol metabolism in the functioning of the retina and possibly in the etiology of ocular diseases, including age-related macular degeneration. PMID- 18385041 TI - Resveratrol inhibits uveal melanoma tumor growth via early mitochondrial dysfunction. AB - PURPOSE: To test the efficacy of resveratrol, a nontoxic plant product, in the treatment of uveal melanoma. METHODS: The effect of oral administration and peritumor injection of resveratrol was tested on tumor growth in two animal models of uveal melanoma. The mechanism of resveratrol action on uveal melanoma cells was studied in vitro in a cell-viability assay: with JC-1 dye, to measure mitochondrial membrane potential; by Western blot analysis, to analyze the cellular redistribution of cytochrome c and Smac/diablo; and in a fluorescence assay with specific substrates, to measure activation of different caspases. RESULTS: Resveratrol treatment inhibited tumor growth in animal models of uveal melanoma. Since oral administration resulted in relatively low bioavailability of resveratrol, the effect of increased local levels was tested by peritumor injection of the drug. This method resulted in tumor cell death and tumor regression. In vitro experiments with multiple uveal melanoma cell lines demonstrate that resveratrol causes a decrease in cell viability, resulting at least in part from an increase in apoptosis through a mitochondrial pathway. An early event in drug action is the direct targeting of mitochondria by resveratrol, which leads to a decrease in mitochondrial membrane potential and the eventual activation of caspase-3. CONCLUSION: These data suggest that resveratrol can inhibit tumor growth and can induce apoptosis via the intrinsic mitochondrial pathway and that by further increasing bioavailability of resveratrol the potency of the drug can be increased, leading to tumor regression. The nontoxic nature of the drug at levels needed for therapy make resveratrol an attractive candidate for the treatment of uveal melanoma. PMID- 18385043 TI - New type of mutations in three spanish families with choroideremia. AB - PURPOSE: Choroideremia (CHM) is an X-linked ophthalmic disease. The gene associated with CHM (REP-1) encodes a ubiquitously expressed protein that is indispensable for the posttranslational activation of retina-specific Rab protein. Different mutations, including large genomic rearrangements involving the REP-1 gene, are responsible for CHM, but they all cause the protein to be truncated or absent. The authors screened 20 Spanish families with clinical diagnoses of CHM to determine the molecular cause of the disease. METHODS: First, the authors performed haplotype analyses to determine whether the disease is linked to the REP-1 gene. In families in whom the disease segregated with the CHM locus (n = 14), mutational screening of the REP-1 gene was performed. RESULTS: In 13 of the 14 families in which the phenotype segregated with the CHM locus, the authors identified the mutation associated with the disease. Eight different molecular defects that led to truncation and one that led to complete absence of the REP-1 protein were found in nine families and one family, respectively. Furthermore, the authors identified a novel type of mutation in the REP-1 gene in three families. This novel type of mutation did not result in a truncated or absent protein. Rather, these patients lost different parts of the REP-1 mRNA in frame that in all the cases encode a conserved protein domain implicated in the interaction with Rab proteins. CONCLUSIONS: Based on the different mutations found, the authors propose a four-step protocol for the molecular diagnosis of CHM. PMID- 18385044 TI - Does the level of physical activity in university students influence development and progression of myopia?--a 2-year prospective cohort study. AB - PURPOSE: To study whether physical activity has a protective effect on the development and progression of myopia in medical students. METHODS: In a 2-year longitudinal cohort study, 156 Caucasian first-year medical students from the University of Copenhagen were enrolled. The baseline examination included visual acuity, subjective refraction, Maddox Wing test (Clement Clarke International Ltd., Harlow, UK), partial coherence interferometry, slit lamp examination, automated refraction in cycloplegia, an oral questionnaire, and a cycle ergometer test. Measurements were repeated at the follow-up. A total of 151 (97%) participants completed the study. RESULTS: The prevalence of myopia (spherical equivalent [SE] 0.20). CONCLUSIONS: The frequency of G6PD deficiency in patients with NAION was significantly lower than expected. Results suggest that G6PD-deficient patients in the Sardinian population have a significantly decreased risk of having NAION. PMID- 18385046 TI - Impact of glaucoma, lens opacities, and cataract surgery on visual functioning and related quality of life: the Barbados Eye Studies. AB - PURPOSE: To determine the relationship of open-angle glaucoma (OAG) and lens opacities to visual functioning and related quality of life (QOL), by using the 25-item National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) in a population of African origin. METHODS: The study included 962 black participants of the Barbados Eye Studies with known glaucoma, prior cataract surgery, visual acuity (VA) or= 0.88, P T point mutation was identified at cDNA position 96, resulting in an R32Q amino acid exchange in a transmembrane domain. The mutation leads to a loss of an SsiI restriction site, which is present in five wild-type mouse strains (102, C3H, C57BL/6, DBA, and JF1). The gene is expressed in the posterior part of the lens vesicle, where the primary fiber elongation starts. In the mutants, the expression pattern of Pax6, Prox1, Six3, and Crygd are modified, but not the pattern of Pax2. CONCLUSIONS: The mutated mouse gene belongs to the family of connexin-encoding genes (gene symbols Gja-Gje). Together with its rat and human homologues, it defines a new subgroup, referred to as Gjf1. The mouse mutant described herein offers a new functional candidate gene for microphthalmia related disorders at the corresponding locus on human chromosome 6, area q24. PMID- 18385073 TI - Novel allele of crybb2 in the mouse and its expression in the brain. AB - PURPOSE: O377 was identified as a new dominant cataract mutation in mice after radiation experiments. The purpose of this study was to genetically characterize the mutation and to analyze its biological consequences. METHODS: Linkage analysis of the O377 mouse mutant was performed; candidate genes including Crybb2 were sequenced. The authors analyzed eyes and brains of the mutants by histology and the expression domains of Crybb2 by in situ hybridization and immunohistochemistry. RNA was isolated from whole brains of heterozygous and homozygous O377 mutants, and differential expression arrays were performed. All studies were compared with age- and strain-matched wild-type mice. RESULTS: The mutation was mapped to chromosome 5 and characterized as an A-->T substitution at the end of intron 5 of the Crybb2 gene. It led to alternative splicing with a 57 bp insertion in the mRNA and to 19 additional amino acids in the protein. In the brain, betaB2-crystallin was expressed in the cerebellum, olfactory bulb, cerebral cortex, and hippocampus. The only morphologic difference in the brain is the increased number of Purkinje cells in the cerebellum of homozygous strain matched mutants. Differential expression analysis revealed the upregulation of calpain-3 in the brain of homozygous mutants, which was confirmed by quantitative real-time PCR. CONCLUSIONS: These results confirm the third allele of Crybb2 in the mouse that also affected exon 6 and the fourth Greek key motif. Moreover, expression analysis of Crybb2 identified for the first time distinct regions of expression in the brain, and the differential expression analysis points to the participation of Ca2+ in the corresponding pathologic processes. PMID- 18385075 TI - Posttranslational modifications in lens fiber connexins identified by off-line HPLC MALDI-quadrupole time-of-flight mass spectrometry. AB - PURPOSE: Gap junction intercellular communication is necessary for the development and maintenance of the lens. Lens fiber connexins are known to be posttranslationally modified, but little detail is available regarding the nature of some of these modifications and the specific amino acids affected. The purpose of this study was to identify posttranslational modification in the bovine lens fiber connexins, Cx44 and Cx49. METHODS: Crude preparations of bovine lens membranes were isolated by centrifugation. The membrane preparations were digested with trypsin or chymotrypsin, and the entire mixture of peptides produced was separated by reverse-phase high performance liquid chromatography and then analyzed by mass spectrometry and tandem mass spectrometry. RESULTS: Coverage of significant portions of the cytoplasmic domains of both Cx44 and Cx49 were successfully obtained. Several of the Ser and Thr residues in the C-tail of Cx44 were phosphorylated, whereas in Cx49 only Ser phosphorylation was detected; however, in this connexin, the phosphorylated residues were located in both the C tail and the central cytoplasmic loop. The data also show that the N-terminal Met residue in each connexin is removed and that the newly exposed N termini become acetylated. In addition, cleavage sites were identified in both proteins. CONCLUSIONS: The study documented the nature and locations of several previously unknown posttranslational modifications in lens fiber connexins. This detailed knowledge of the specific posttranslationally modified sites will allow further work to elucidate the mechanisms that different signaling pathways use to regulate connexins in lens fiber cells. PMID- 18385074 TI - Dual roles for Prox1 in the regulation of the chicken betaB1-crystallin promoter. AB - PURPOSE: Lens fiber cell differentiation is marked by the onset of betaB1 crystallin expression and is controlled by the cooperative action of a set of transcription factors including Prox1, an atypical homeodomain protein. Previously, the authors reported that Prox1 directly interacts with the OL2 element found in the chicken betaB1-crystallin basal promoter to activate the expression of this gene. Here they mapped the location of activating and repressing sequences of the full-length chicken betaB1-crystallin promoter ( 432/+30) in lens epithelial cells, annular pad cells, and intact lens and characterized Prox1-binding sites found in this region. METHODS: Transfection analysis and transgenic mice were used to characterize upstream regions of the chicken betaB1-crystallin gene. DNaseI footprinting and chromatin immunoprecipitation was performed to identify Prox1-binding sites, and transfection analyses were used to characterize these sites functionally. RESULTS: Sequences between -152 and -432 of the chicken betaB1-crystallin promoter mediated either promoter activation or repression, depending on the stage of lens differentiation tested. Two new Prox1-binding sites were found in this region that bound Prox1 more avidly than the OL2 element. However, neither binding site conferred Prox1-mediated activation on a heterologous promoter; instead, each allowed Prox1 to repress promoter function. CONCLUSIONS: The function of the upstream region of the chicken betaB1-crystallin promoter changes depending on cellular context. These data suggest that Prox1 function as a transcriptional activator could be regulated at the DNA level based on the characteristics of the responsive elements. PMID- 18385076 TI - Bestrophin-2 is involved in the generation of intraocular pressure. AB - PURPOSE: The bestrophin family of proteins has been demonstrated to generate or regulate Ca2+-activated Cl(-) conductances. Mutations in bestrophin-1 (Best1) cause several blinding eye diseases, but little is known about other bestrophin family members. This study involved disruption of the Best2 gene in mice. METHODS: The mouse Best2 gene was disrupted by replacing exons 1, 2, and part of exon 3 with a Lac Z. The expression profile of Bestrophin-2 (Best2) was examined using RT-PCR, X-gal staining, and immunohistochemistry. Intraocular pressure (IOP) was measured by anterior chamber cannulation. RESULTS: RT-PCR of mouse tissues revealed Best2 mRNA in eye, colon, nasal epithelia, trachea, brain, lung, and kidney. X-gal staining, confirmed expression in colon epithelia and in the eye, in the nonpigmented epithelia (NPE). Best2 was not expressed in RPE cells. Best2 protein was observed only in NPE and colon epithelia. The absence of Best2 had no obvious deleterious effect on the mice. However, the Best2-/- mice were found to have significantly (P < 0.02) diminished IOP with respect to the Best2+/+ and Best2+/- littermates. The Best2-/- and Best2+/- mice responded better to the carbonic anhydrase inhibitor brinzolamide than did their Best2+/+ littermates, although the beta-blocker timolol brought IOP to the same level, regardless of genotype. CONCLUSIONS: Best2 plays a role in the generation of IOP by regulating formation of aqueous humor, and inhibition of Best2 function represents an attractive new avenue for regulating IOP in individuals with glaucoma. PMID- 18385077 TI - Characterization of outer retinal morphology with high-speed, ultrahigh resolution optical coherence tomography. AB - PURPOSE: To visualize, quantitatively assess, and interpret outer retinal morphology by using high-speed, ultrahigh-resolution (UHR) OCT. METHODS: Retinal imaging was performed in the ophthalmic clinic in a cross-section of 43 normal subjects with a 3.5-microm, axial-resolution, high-speed, UHR OCT prototype instrument, using a radial scan pattern (24 images, 1500 axial scans). Outer retinal layers were automatically segmented and measured. High-definition imaging was performed with a 2.8-microm axial-resolution, high-speed, UHR OCT research prototype instrument, to visualize the finer features in the outer retina. RESULTS: Quantitative maps of outer retinal layers showed clear differences between the cone-dominated fovea and the rod-dominated parafovea and perifovea, indicating that photoreceptor morphology can explain the appearance of the outer retina in high-speed, UHR OCT images. Finer, scattering bands were visualized in the outer retina using high-definition imaging and were interpreted by comparison to known anatomy. CONCLUSIONS: High-speed UHR OCT enables quantification of scattering layers in the outer retina. An interpretation of these features is presented and supported by quantitative measurements in normal subjects and comparison with known anatomy. The thick scattering region of the outer retina previously attributed to the retinal pigment epithelium (RPE) is shown to consist of distinct scattering bands corresponding to the photoreceptor outer segment tips, RPE, and Bruch's membrane. These results may advance understanding of the outer retinal appearance in OCT images. The normative measurements may also aid in future investigations of outer retinal changes in age-related macular degeneration and other diseases. PMID- 18385078 TI - Retinal laminar architecture in human retinitis pigmentosa caused by Rhodopsin gene mutations. AB - PURPOSE: To determine the underlying retinal micropathology in subclasses of autosomal dominant retinitis pigmentosa (ADRP) caused by rhodopsin (RHO) mutations. METHODS: Patients with RHO-ADRP (n = 17, ages 6-73 years), representing class A (R135W and P347L) and class B (P23H, T58R, and G106R) functional phenotypes, were studied with optical coherence tomography (OCT), and colocalized visual thresholds were determined by dark- and light-adapted chromatic perimetry. Autofluorescence imaging was performed with near-infrared light. Retinal histology in hT17M-rhodopsin mice was compared with the human results. RESULTS: Class A patients had only cone-mediated vision. The outer nuclear layer (ONL) thinned with eccentricity and was not detectable within 3 to 4 mm of the fovea. Scotomatous extracentral retina showed loss of ONL, thickening of the inner retina, and demelanization of RPE. Class B patients had superior inferior asymmetry in function and structure. The superior retina could have normal rod and cone vision, normal lamination (including ONL) and autofluorescence of the RPE melanin; laminopathy was found in the scotomas. With Fourier-domain-OCT, there was apparent inner nuclear layer (INL) thickening in regions with ONL thinning. Retinal regions without ONL had a thick hyporeflective layer that was continuous with the INL from neighboring regions with normal lamination. Transgenic mice had many of the laminar abnormalities found in patients. CONCLUSIONS: Retinal laminar abnormalities were present in both classes of RHO-ADRP and were related to the severity of colocalized vision loss. The results in human class B and the transgenic mice support the following disease sequence: ONL diminution with INL thickening; amalgamation of residual ONL with the thickened INL; and progressive retinal remodeling with eventual thinning. PMID- 18385080 TI - Retinal microglia and uveal tract dendritic cells and macrophages are not CX3CR1 dependent in their recruitment and distribution in the young mouse eye. AB - PURPOSE: The chemokine receptor CX3CR1 is expressed by monocyte-derived dendritic cells (DCs) and macrophages. CX3CR1 mediates leukocyte migration and adhesion in homeostatic and inflammatory conditions. Mice lacking Cx3cr1 have altered distribution and function of DC subpopulations in some tissue microenvironments. The present study compares the distribution of monocyte-derived cells in the normal retina and uveal tract as a prelude to the investigation of the role of CX3CR1 in murine models of ocular disease. METHODS: Transgenic mice in which either one (Cx3cr1 gfp/+, heterozygous) or both (Cx3cr1 gfp/gfp, homozygous) copies of the Cx3cr1 gene have been replaced by the enhanced green fluorescent protein (eGFP) reporter gene were used to investigate the role of Cx3cr1 expression on macrophages and DCs in the normal uveal tract and retina. Chimeric mice were used to investigate turnover of these cells in the normal, uninflamed eye. RESULTS: Confocal analysis found no significant differences in the density, phenotype or morphology of eGFP+ cells between Cx3cr1 gfp/+ and Cx3cr1 gfp/+ mice in immunostained iris, ciliary body, or choroidal and retinal wholemounts. Flow cytometry also failed to detect any difference in the density or cell shape of eGFP+ cells between Cx3cr1 gfp/+ and Cx3cr1 gfp/+ mice. Chimeras revealed 73% turnover of monocyte-derived cells in the iris and 63% in the choroid by 6 weeks after transplantation. CONCLUSIONS: These data illustrate that homing or migration of DCs and macrophages to the uveal tract and retina in normal young mice is not Cx3cr1 dependent and provide a solid foundation for future studies of monocyte-derived cells and the role of Cx3cr1 in models of ocular disease. PMID- 18385079 TI - Activated NAD(P)H oxidase from supplemental oxygen induces neovascularization independent of VEGF in retinopathy of prematurity model. AB - PURPOSE: To study NAD(P)H oxidase-dependent outcomes after oxygen stresses that are similar to those experienced by preterm infants today using a rat model of retinopathy of prematurity. METHODS: Within 4 hours of birth, pups and their mothers were cycled between 50% and 10% oxygen daily for 14 days and were returned to room air (21% O2, 50/10 oxygen-induced retinopathy [OIR]) or supplemental oxygen (28% O2, 50/10 OIR+SO) for 4 days. Pups received intraperitoneal injections of the specific NAD(P)H oxidase inhibitor apocynin (10 mg/kg/d) or of PBS from postnatal day (P)12 to P17, and some received intraperitoneal injections of hypoxyprobe before kill. Intravitreous neovascularization (IVNV), avascular/total retinal areas, vascular endothelial growth factor (VEGF), NAD(P)H oxidase activity, or hypoxic retina (conjugated hypoxyprobe) were determined in neurosensory retinas. Human retinal microvascular endothelial cells (RMVECs) treated with apocynin or control were exposed to 1% or 21% O2 and assayed for phosphorylated (p-)Janus kinase (JNK) and NAD(P)H oxidase activity. RESULTS: Retinas from 50/10 OIR+SO had increased NAD(P)H oxidase activity and lower VEGF than did retinas from 50/10 OIR. Apocynin treatment reduced the IVNV area and hypoxic retina in 50/10 OIR+SO. RMVECs treated with 1% O2 had increased p-JNK compared with RMVECs exposed to room air. CONCLUSIONS: Different oxygen stresses activate NAD(P)H oxidase to varying degrees to trigger disparate pathways (angiogenesis or apoptosis). The oxygen stresses and outcomes used in this study are relevant to human ROP and may explain some of the complexity in the pathophysiology of ROP resulting from oxygen exposure. PMID- 18385081 TI - Impact of optic media opacities and image compression on quantitative analysis of optical coherence tomography. AB - PURPOSE: To analyze the impact of opacities in the optical pathway and image compression of 32-bit raw data to 8-bit jpg images on quantified optical coherence tomography (OCT) image analysis. METHODS: In 18 eyes of nine healthy subjects, OCT images were acquired from the central macula. To simulate opacities in the optical system, neutral-density (ND) filters with linear absorption spectra were placed between the OCT device and examined eyes. Light reflection profiles (LRPs) of images acquired with various ND filters were compared. LRPs of the 32-bit raw data were compared with those obtained from the 8-bit jpg compressed images. RESULTS: ND filters induced a linear decrease of reflectivity in OCT images, depending on initial signal intensity. Quantitative OCT analysis showed no significant difference between 32-bit raw data and 8-bit jpg files (P > 0.05). CONCLUSIONS: Quantitative OCT analysis is not significantly influenced by data compression. A mathematical model can correct for optical opacities to improve OCT images. PMID- 18385083 TI - Local diabetic retinopathy prediction by multifocal ERG delays over 3 years. AB - PURPOSE: To derive and validate a model for use in predicting local retinal areas in which nonproliferative diabetic retinopathy (NPDR) lesions will develop over a 3-year period, by using primarily the implicit time (IT) of the multifocal electroretinogram (mfERG). METHODS: Eighteen diabetic patients were examined at baseline and at three annual follow-ups. Ophthalmic examinations, including fundus photographs and mfERG testing, were performed at each visit. Thirty-five retinal zones were constructed from the 103-element stimulus array, and each zone was assigned the maximum IT z-score within it based on 30 age-similar control subjects. Logistic regression was used to investigate the development of retinopathy in relation to baseline mfERG IT delays and additional diabetic health variables. Receiver operating characteristic (ROC) curves were used to evaluate the models. RESULTS: Retinopathy developed in 77 of the 1208 retinal zones, of which 25 had recurring retinopathy. Multivariate analyses yielded baseline mfERG IT, duration of diabetes, and blood glucose concentration as the most important predictors of recurring retinopathy. mfERG ITs were not predictive of transient retinopathy. ROC curves based on the multivariate model for the prediction of recurring retinopathy resulted in an area under the curve of 0.95, sensitivity of 88%, and specificity of 98%. Ten-fold cross-validation confirmed the high sensitivity and specificity of the model. CONCLUSIONS: The development of recurring retinopathy over a 3-year period can be well predicted by using a multivariate model based on mfERG implicit time. Multifocal ERG delays are promising candidate measures for trials of novel therapeutics directed at preventing or slowing the progression of NPDR. PMID- 18385082 TI - Circadian regulation of retinoschisin in the chick retina. AB - PURPOSE: To investigate the circadian regulation and acute illumination effects on the expression and secretion of retinoschisin from vertebrate retinas. METHODS: Retinas were studied on the second day of constant darkness (DD) after several days of entrainment to 12-hour light/12-hour dark (LD) cycles in ovo or in vitro. Quantitative real-time PCR and Western immunoblotting were used to examine the mRNA and protein expressions of retinoschisin at different circadian time points. Pharmacologic treatments in whole retina and dissociated retinal cell cultures were used to investigate the cellular mechanisms underlying the circadian regulation of retinoschisin content and secretion. Different illumination conditions were given to examine changes in retinoschisin content in association with acute light/dark adaptation. RESULTS: The mRNA level, protein expression, and secretion of retinoschisin were under circadian control, all of which were higher at night and lower during the day. The Ras, MAP kinase Erk, CaMKII pathway served as part of the circadian output regulating the rhythmicity of retinoschisin. Blockage of L-type VGCCs dampened the retinoschisin rhythm, but inhibition of L-type VGCCs did not completely abolish the secretion of retinoschisin. The protein expression of retinoschisin also responded to acute illumination changes. CONCLUSIONS: The mRNA and protein expression, as well as retinoschisin secretion, are under circadian control. L-type VGCCs play a role in the circadian regulation of retinoschisin, but the molecular mechanism underlying retinoschisin secretion does not depend on L-type VGCCs. Protein expression of retinoschisin in response to acute illumination changes depends on previous light exposure experience. PMID- 18385084 TI - Threshold amplitude and frequency for ocular tissue release from a vibrating instrument: an experimental study. AB - PURPOSE: During retinal pigment epithelium (RPE) and choroid graft translocation in the treatment of patients with exudative age-related macular degeneration, the adhesion of the graft to the translocation instrument complicated its submacular release. Vibration of the instrument improved the release of the graft. This study was conducted to validate the effectiveness of the principle of vibration and to determine the threshold amplitude and frequency required for development of an optimized instrument. METHODS: An experimental in vitro model with fresh porcine RPE-choroid grafts was used. Release of the graft was studied by a masked observer for amplitudes in the range of 0.05 to 1.2 mm and frequencies in the range of 25 to 200 Hz in the horizontal plane. RESULTS: The minimum threshold amplitude required to release the graft was approximately 0.15 mm from a frequency of 100 Hz and higher. CONCLUSIONS: This study confirmed the clinical experience that vibration of an instrument induces the release of the RPE-choroid graft. The minimum threshold amplitude and frequency needed for optimum tissue release were estimated. PMID- 18385085 TI - Evaluation of the new photosensitizer Stakel (WST-11) for photodynamic choroidal vessel occlusion in rabbit and rat eyes. AB - PURPOSE: To evaluate the photodynamic potential of a new hydrosoluble photosensitizer (WST-11, Stakel; Steba Biotech, Toussus-Le-Noble, France), for use in occlusion of normal choroidal vessels in the rabbit eye and CNV (choroidal neovascularization) in the rat eye. METHODS: Occlusive and nonocclusive parameters of Stakel and verteporfin photodynamic therapy (PDT) were investigated in pigmented rabbits. Eyes were followed by fluorescein angiography (FA) and histology at various intervals after PDT. RESULTS: When occlusive parameters (fluence of 50 J/cm(2), 5 mg/kg drug dose and DLI [distance to light illumination] of 1 minute) were used, Stakel PDT was efficient immediately after treatment without associated structural damage of the RPE and retina overlying the treated choroid in the rabbit eye. Two days later, total occlusion of the choriocapillaries was seen in 100% of the treated eyes, along with accompanying histologic structural changes in the overlying retina. When the occlusive parameters (fluence, 100 J/cm2; drug dose, 12 mg/m2; and DLI, 5 minutes) of verteporfin PDT were used, occlusion of the choriocapillaries was observed in 89% of the treated eyes. Histology performed immediately after treatment demonstrated structural damage of the overlying retina and RPE layer. Weaker, nonocclusive Stakel PDT parameters (25 J/cm2, 5 mg/kg, and DLI of 10 minutes) did not induce choriocapillary occlusion or retinal lesions on FA or histology. Weaker, nonocclusive verteporfin PDT parameters (10 J/cm2, 0.2 mg/kg, and DLI of 5 minutes) did not induce choriocapillary occlusion. However, histology of these eyes showed the presence of damage in the retinal and choroidal tissues. Moreover, preliminary results indicate that selective CNV occlusion can be achieved with Stakel PDT in the rat eye. CONCLUSIONS: Unlike verteporfin PDT, Stakel PDT does not cause direct damage to the RPE cell layer or retina. These observations indicate that Stakel PDT may have a high potential for beneficial therapeutic outcomes in treatment of AMD. PMID- 18385086 TI - Beneficial effect of zeaxanthin on retinal metabolic abnormalities in diabetic rats. AB - PURPOSE: Oxidative damage and growth factors are implicated in the pathogenesis of retinopathy in diabetes. Recent studies have shown that two dietary carotenoids, lutein and zeaxanthin (Zx), that are specifically concentrated within ocular tissues, may play important roles in maintaining their integrity. This study is to evaluate the potential protective effects of Zx against retinal oxidative damage and growth factors in diabetes. METHODS: A group of rats received normal powdered diet or powdered diet supplemented with 0.02% or 0.1% Zx soon after induction of diabetes. Age-matched normal rats served as control subjects. At 2 months of diabetes, oxidative stress, vascular endothelial cell growth factor (VEGF), and intercellular adhesion molecule (ICAM)-1 were quantified in the retina. RESULTS: Zx supplementation prevented diabetes-induced increase in retinal damage, and increases in VEGF and ICAM-1. The levels of lipid peroxide, oxidatively modified DNA, electron transport complex III, nitrotyrosine, and mitochondrial superoxide dismutase were similar in the retinas of Zx-treated diabetic rats and normal control rats, and these values were significantly different from those obtained from diabetic rats without any supplementation. In the same rats, Zx also prevented diabetes-induced increases in retinal VEGF and ICAM-1. Both 0.02% and 0.1% Zx had similar effects on diabetes-induced retinal abnormalities, and these effects were achieved without ameliorating the severity of hyperglycemia. However, Zx administration failed to prevent a diabetes-induced decrease in retinal GSH levels. CONCLUSIONS: Zx significantly inhibits diabetes-induced retinal oxidative damage and elevation in VEGF and adhesion molecule, all abnormalities that are associated with the pathogenesis of diabetic retinopathy. The results suggest that Zx supplementation has the potential to inhibit the development of retinopathy in diabetic patients. PMID- 18385087 TI - Toll-like receptor polymorphisms and age-related macular degeneration. AB - PURPOSE: Evidence from genetic-association studies in conjunction with the demonstration of complement deposition in the retina and choroid implicates noncellular pathways of innate immunity in the pathogenesis of age-related macular degeneration (AMD). The purpose of this study was to determine whether common variation in the 10 human toll-like receptors (TLRs) alters the risk of AMD. METHODS: Sixty-eight SNPs were iteratively genotyped across the TLR genes in a cohort of 577 subjects, with and without AMD. Two additional cohorts were used for replication studies. Standard genetic-association methods were used to analyze the results for association with disease and interaction with other loci. RESULTS: Coding SNPs in TLR3 (rs3775291) and TLR7 (rs179008) showed association with AMD in one group (P = 0.01 and P = 0.02, respectively) before correction for multiple testing. For both SNPs, the association with AMD arose due to an excess of heterozygotes compared with homozygotes for the major allele. The two coding SNPs were not associated with AMD in another case-control cohort or an extended family cohort. Although an intronic SNP in TLR4 was associated marginally with AMD (P = 0.03), it was not possible to replicate a previous association with the rare coding SNP D299G in this gene (P = 0.6). CONCLUSIONS: Although borderline support for association between polymorphisms in TLR genes and AMD was reported for some cohorts, these initial observations of coding SNPs in TLR3, TLR4, and TLR7 were not replicated. TLR variants are unlikely to have a major impact on overall AMD risk, and the common variants studied were not associated with AMD. PMID- 18385088 TI - Carnitine palmitoyltransferase I and Acyl-CoA dehydrogenase 9 in retina: insights of retinopathy in mitochondrial trifunctional protein defects. AB - PURPOSE: Progressive pigment chorioretinopathy is a major long-term complication of mitochondrial trifunctional protein (MTP) defects, disorders of mitochondrial fatty acid beta-oxidation. To better understand the pathogenesis of the retinopathy component, the authors studied expression of the main regulatory protein of the beta-oxidation pathway, carnitine palmitoyltransferase (CPT) 1, and acyl-CoA dehydrogenase (ACAD) 9 in retinal sections and cultured cells. METHODS: Immunoblotting was performed with polyclonal antibodies to ACAD9 and the three isoforms of CPT1. In quantitative real-time PCR (QRT-PCR), predesigned gene specific probes and primer sets for human CPT1 isoforms were used. In situ hybridization (ISH) and immunohistochemistry was performed on formalin-fixed, paraffin-embedded sections of the rat and human eye. RESULTS: The predominant CPT1 mRNA types detected by QRT-PCR in cultured human retinal pigment epithelial cells were of the liver (CPT1A) and brain (CPT1C) isotypes. CPT1A and ACAD9 protein expression was found in cultured human and rat RPE and rat neural retinal precursor cells. ISH of rat retinal sections showed CPT1A and CPT1C expression in the retinal pigment epithelium (RPE), the inner nuclear layer, and the ganglion cell layer. CPT1A expression was also detected in the Muller cell microvilli, and CPT1C expression was detected in the photoreceptor inner segments. ACAD9 immunolabeling was detected in rat and human RPE, human photoreceptor inner segments, and ganglion cell layer. CONCLUSIONS: These findings imply that the mitochondrial fatty acid beta-oxidation pathway probably is active in metabolism of the RPE and certain neuroretinal cell types. Accumulation of 3-hydroxylated intermediates of long-chain fatty acids may contribute to the pathogenesis of retinopathy in MTP deficiencies. PMID- 18385089 TI - Modulation of Nrf2-dependent antioxidant functions in the RPE by Zip2, a zinc transporter protein. AB - PURPOSE: To characterize the involvement of Zip2, a zinc transporter protein, in the antioxidant functions of cultured human retinal pigment epithelial (RPE) cells. METHODS: The expression of zinc transporter proteins was determined by RT PCR. Intracellular zinc concentration was assessed by staining with a zinc sensitive dye followed by flow cytometry. Stable overexpression of the transporter protein Zip2 was achieved by transducing ARPE-19 cells with a retroviral vector containing the open reading frame of the human Zip2 gene. Activity of nuclear factor erythroid 2-related factor 2 (Nrf2) was measured using a dual luciferase assay after transient transfection of reporter plasmids containing the antioxidant response element (ARE). Glutamate-cysteine ligase (GCL) expression was measured by quantitative real-time RT-PCR. RESULTS: Cultured RPE cells could transport zinc with Zip2 as an influx transporter expressed in ARPE-19 cells and human RPE cells isolated from postmortem donor eyes. The mRNA level of Zip2 was influenced by intracellular and extracellular zinc concentrations. Overexpression of Zip2 resulted in increased Nrf2 activity, higher GCL expression, and increased glutathione synthesis. CONCLUSIONS: RPE cells can actively uptake zinc through the transporter Zip2, and the increased intracellular zinc upregulates the Nrf2-dependent antioxidant function. PMID- 18385091 TI - Inhibition of choroidal neovascularization with an anti-inflammatory carotenoid astaxanthin. AB - PURPOSE: Astaxanthin (AST) is a carotenoid found in marine animals and vegetables. The purpose of the present study was to investigate the effect of AST on the development of experimental choroidal neovascularization (CNV) with underlying cellular and molecular mechanisms. METHODS: Laser photocoagulation was used to induce CNV in C57BL/6J mice. Mice were pretreated with intraperitoneal injections of AST daily for 3 days before photocoagulation, and treatments were continued daily until the end of the study. CNV response was analyzed by volumetric measurements 1 week after laser injury. Retinal pigment epithelium choroid levels of IkappaB-alpha, intercellular adhesion molecule (ICAM)-1, monocyte chemotactic protein (MCP)-1, interleukin (IL)-6, vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR)-1, and VEGFR-2 were examined by Western blotting or ELISA. AST was applied to capillary endothelial (b-End3) cells, macrophages, and RPE cells to analyze the activation of NF-kappaB and the expression of inflammatory molecules. RESULTS: The index of CNV volume was significantly suppressed by treatment with AST compared with that in vehicle treated animals. AST treatment led to significant inhibition of macrophage infiltration into CNV and of the in vivo and in vitro expression of inflammation related molecules, including VEGF, IL-6, ICAM-1, MCP-1, VEGFR-1, and VEGFR-2. Importantly, AST suppressed the activation of the NF-kappaB pathway, including IkappaB-alpha degradation and p65 nuclear translocation. CONCLUSIONS: AST treatment, together with inflammatory processes including NF-kappaB activation, subsequent upregulation of inflammatory molecules, and macrophage infiltration, led to significant suppression of CNV development. The present study suggests the possibility of AST supplementation as a therapeutic strategy to suppress CNV associated with AMD. PMID- 18385090 TI - Essential roles of the PI3 kinase/Akt pathway in regulating Nrf2-dependent antioxidant functions in the RPE. AB - PURPOSE: To investigate functional interactions between the phosphatidylinositol 3-kinase (PI3K)/Akt pathway and the nuclear factor erythroid 2-related factor 2 (Nrf2)-dependent antioxidant system in cultured human retinal pigment epithelium (RPE) cells. METHODS: Cultured ARPE-19 cells were treated with different concentrations of PI3K inhibitors, followed by exposure to sulforaphane, an Nrf2 inducer. Akt phosphorylation was detected by Western blot analysis. Intracellular glutathione (GSH) content was measured by HPLC. Expression of genes downstream of Nrf2, including glutamate-cysteine ligase (GCL) and glutathione S-transferase, was measured by quantitative RT-PCR. Nrf2 activity was measured by a dual luciferase assay after transfection of a reporter plasmid containing the antioxidant response element (ARE). The small interference RNA approach was used to knock down Nrf2 in the RPE. Nrf2 localization was determined by subcellular fractionation and Western blot analyses. RESULTS: PI3K inhibitors wortmannin and LY294002 caused dose-dependent cellular and mitochondrial GSH depletion and downregulation of the modulatory subunit of GCL in cultured RPE cells. Both the basal and the induced Nrf2 activities were inhibited by wortmannin and LY294002. Overexpression of a constitutively active form of Akt potentiated Nrf2 activation, and the effect of Akt was blocked by siRNA that knocked down Nrf2. LY294002 also inhibited sulforaphane-induced Nrf2 nuclear translocation. CONCLUSIONS: The PI3K/Akt pathway plays key roles in regulating Nrf2-ARE dependent protection against oxidative stress in the RPE. PMID- 18385092 TI - Elevated MMP Expression in the MRL Mouse Retina Creates a Permissive Environment for Retinal Regeneration. AB - PURPOSE: The MRL/MpJ (healer) mouse is an established model for autoimmune studies and was recently identified as having a profound ability to undergo scarless regeneration of the tissue in the ear and heart. This regenerative capacity has been linked to elevated matrix metalloproteinase (MMP)-2 and -9 expression, giving this mouse the ability to degrade and remove inhibitory basement membrane molecules. Although elevated MMP expression has been reported in somatic tissues in this strain, little is known about MMP expression and the response to injury in the MRL/MpJ mouse retina. The purpose of this study was to investigate whether increased MMP expression and subsequent decreased inhibitory extracellular matrix molecule deposition in the MRL/MpJ mouse retina produces a permissive regenerative environment. METHODS: Experiments were performed using 3- to 4-week-old MRL/MpJ, retinal degenerative (rd1), and C57BL/6 (wild-type) mice. Western blotting, oligo-microarray, and immunohistochemical analyses were used to determine the level and location of MMP and extracellular matrix (ECM) protein expression. Retinal responses to injury were modeled by retinal detachment in vivo and in retinal explantation in vitro. The capacity of the retinal environment to support photoreceptor cell migration, integration, or regeneration was analyzed using hematoxylin-eosin, immunohistochemical staining, and cell counting. RESULTS: Compared with C57BL/6J animals, MRL/MpJ mice exhibit elevated levels of MMP-2, -9, and -14 and decreased levels of the inhibitory proteins neurocan and CD44 within the retina. Although similar increases in MMP-2, -9, and CD44s (CD44 degradation product) were observed in the rd1 retina, elevated levels of the inhibitory ECM molecules (neurocan and CD44) remained. Thus, the MRL retinal environment, which expresses lower levels of inhibitory ECM molecules after injury, was more conducive to regeneration and enhanced photoreceptor integration in vitro than C57BL/6J or rd1 controls. CONCLUSIONS: The MRL mouse retina shows elevated MMP expression and decreased levels of scar-related inhibitory molecules, which leads to a retinal environment that is more permissive for neural regeneration and cell integration after in vitro retinal explantation. PMID- 18385093 TI - FGF19 exhibits neuroprotective effects on adult mammalian photoreceptors in vitro. AB - PURPOSE: Several fibroblast growth factors (FGFs) exhibit neuroprotective influences against retinal photoreceptor degeneration. The expression of FGF receptor (FGFR) 4 on photoreceptors suggests a specific ligand, FGF-19, might also be beneficial. The authors hence examined the potential role of FGF-19 in this regard. METHODS: Adult human retinal sections were processed for anti-FGFR-4 immunohistochemistry. Total RNA and proteins were extracted from parallel cultures of human Y79 retinoblastoma and primary adult pig photoreceptors; RNA samples were used for RT-PCR analysis of FGF-19, and proteins were subjected to immunoprecipitation for FGFR-1 and FGFR-4 or to Western blotting of FGF-19. Cultures were incubated with increasing concentrations of FGF-19 before extraction and Western blotting for phosphotyrosine. Photoreceptor cultures were screened for cell survival and processed for immunocytochemistry using anti neural retina leucine zipper (Nrl) antibody. RESULTS: FGF-19 mRNA was detected in adult pig retinal pigment epithelial cells, and FGF-19 protein was found in cell extracts and conditioned medium prepared from retinal pigment epithelium. The addition of FGF-19 to Y79 retinoblastoma or primary adult pig photoreceptor cultures led to time- and dose-dependent changes in proliferation (for Y79) or survival (for primary photoreceptors). FGF-19 induced the phosphorylation of an FGFR-4-immunoreactive band of approximately 80 kDa and led to the heterodimerization of FGFR-1 and FGFR-4. Y79 and primary photoreceptor cells maintained in serum-supplemented media exhibited Nrl immunoreactivity by Western blotting, which decreased after serum deprivation. The addition of FGF-19 led to the reexpression of Nrl immunoreactivity in both culture models. CONCLUSIONS: These data indicate a physiological role for FGF-19 in adult photoreceptor phenotypic maintenance and survival and argue in favor of its use as a neuroprotectant. PMID- 18385094 TI - Mecamylamine suppresses Basal and nicotine-stimulated choroidal neovascularization. AB - PURPOSE: Nicotinic acetylcholine receptors (nAChR) are best known for their role in neurotransmission, but they have recently been demonstrated on vascular endothelial cells. Acetylcholine is their endogenous ligand, but they are also stimulated by nicotine. By stimulating nAChR, nicotine promotes tumor angiogenesis as well as atherosclerotic plaque neovascularization. In this study, the authors investigated the role of nAChR in the pathogenesis of choroidal neovascularization (CNV). METHODS: The effect of the nonselective nAChR antagonist mecamylamine was tested on human retinal and choroidal endothelial cells in vitro and in a murine model of CNV. RESULTS: Several nAChR isoforms were identified in retinal and choroidal microvascular endothelial cells, and the ability of these cells to form tubules when grown in growth factor-reduced basement membrane matrix and supplemented with VEGF was suppressed by the nAChR antagonist mecamylamine. Supplementation of the drinking water of mice with nicotine increased the size of CNV lesions at Bruch membrane rupture sites, an effect that was blocked by subcutaneous administration of mecamylamine (50 mg/kg/d) by an osmotic pump. In the absence of nicotine, CNV formation was suppressed by the infusion of 50 mg/kg/d mecamylamine or by topical application 0.1 or 1% mecamylamine to the cornea. CONCLUSIONS: These data suggest that endogenous activation of nAChR promotes CNV and that activation of nAChR by nicotine may contribute to the increased incidence of CNV seen in smokers with age-related macular degeneration (AMD). Topically administered mecamylamine could provide an appealing new treatment approach for CNV. PMID- 18385095 TI - The protective effect of quercetin against oxidative stress in the human RPE in vitro. AB - PURPOSE: To investigate the possible protective effect of the dietary antioxidant quercetin on retinal pigment epithelial (RPE) cell dysfunction and cellular senescence occurring in age-related macular degeneration (AMD). The major flavonoid quercetin was studied on RPE cells in vitro. METHODS: Cultured human RPE cells were incubated with different concentrations of quercetin for 24 hours. Cells were then treated with 150 to 300 microM hydrogen peroxide for 2 hours. Mitochondrial function was measured by using MTT assay and cell vitality by live dead staining assay. Intracellular levels of glutathione were determined by using a glutathione assay kit. Apoptosis was quantified by a caspase-3 assay, and cellular senescence was quantified by beta-galactosidase staining. Expression of the senescence-associated transmembrane protein caveolin-1 was investigated by Northern and Western blot analyses. RESULTS: Hydrogen peroxide treatment caused significant decreases in mitochondrial function (52%) and in cell vitality (71%), whereas preincubation with 50 microM quercetin diminished this decrease in a dose dependent manner. Quercetin treatment did not show any notable effect on intracellular levels of glutathione in either used concentration of quercetin. Hydrogen peroxide-induced activation of caspase-3 was reduced by 50 microM quercetin, from 1.9- to 1.4-fold, compared with untreated control (P < 0.001). Hydrogen peroxide caused a large (>90%) dose-dependent increase in beta galactosidase-positive cells, whereas in the untreated control only single cells expressed this enzyme (<5%). This increase in cellular senescence was significantly attenuated by quercetin in a dose-dependent manner. The highest attenuation was reached at 50 microM quercetin. Quercetin caused a significant dose-dependent reduction of caveolin-1 mRNA 48 hours after treatment with hydrogen peroxide. After 96 hours of incubation, caveolin-1 protein levels were also reduced. CONCLUSIONS: The data demonstrate that quercetin is able to protect RPE cells from oxidative damage and cellular senescence in vitro in a dose dependent manner. The authors suggest that this increase in antioxidative capacity is--among other mechanisms, such as the intracellular redox state--also mediated by inhibiting the upregulation of caveolin-1. Downregulation of caveolin 1 may be important for the retinal pigment epithelium to prevent apoptotic cell death in response to cellular stress, a condition implicated in the early pathogenesis of AMD. Therefore, the authors believe that the use of antioxidative dietary flavonoids such as quercetin is a promising approach in the prevention of early AMD. PMID- 18385097 TI - An FMRI study of saccadic and smooth-pursuit eye movement control in patients with age-related macular degeneration. AB - PURPOSE: To compare the cortical networks that underlie oculomotor function in patients with age-related macular degeneration (AMD) with those in normally sighted control subjects, using functional magnetic resonance imaging (fMRI). METHODS: Six patients with bilateral geographic retinal atrophy due to AMD (age range, 55-83 years) were recruited for the study. The visual acuities of the patients ranged from 20/76 (0.58 logMAR) to 20/360 (1.26 logMAR). An additional six younger (age range, 22-31 years) and six older (age range, 54-78 years) normally sighted individuals were recruited as control subjects. fMRI data were acquired on a 3.0-Tesla, scanner while subjects performed visually guided saccade (VGS) and smooth-pursuit (SmP) tasks. RESULTS: Contrasts between VGS and fixation on a stationary target identified a network of activation that included the frontal eye fields (FEFs), supplementary eye fields (SMA/SEFs), prefrontal cortex (PFC), intraparietal sulci (IPS), and the areas of the visual cortex (MT/V5, V2/V3, and V1) in control subjects and patients. A similar network was identified for comparisons between SmP and periods of fixation. Marked variability was observed in the performance of both tasks across all patients. For both tasks, the patients generally showed increased PFC and IPS activation, with decreased activation in visual cortex compared with the control subjects. The patients showed significantly increased activation of the FEFs and SMA/SEFs in the SmP task, compared with the control subjects. CONCLUSIONS: These data suggest that performance of both eye movement tasks required greater involvement of the cortical regions generally implicated in attention and effort in patients with AMD. PMID- 18385096 TI - Characterization of a Bcl-XL-interacting protein FKBP8 and its splice variant in human RPE cells. AB - PURPOSE: The immunophilin protein FKBP8 interacts with Bcl2/Bcl-XL and is essential for mouse eye development. The purpose of this study was to define the expression of the FKBP8 gene in cultured human RPE cells and explore its involvement in the control of apoptosis. METHODS: Rapid amplification of cDNA ends (RACE) was performed on RNA isolated from human RPE cells. The existence of FKBP8 and a splice variant was confirmed by RT-PCR. The interaction between Bcl XL and FKBP8 was measured by coimmunoprecipitation. ARPE-19 cells stably overexpressing FKBP8 and its splice variant were established. Their responses to thapsigargin and t-butyl hydroperoxide-induced cell death were measured by flow cytometry. Apoptosis was determined by terminal deoxyribonucleotidyl transferase mediated fluorescein-dUTP nick-end labeling (TUNEL) assay. The activities of the nuclear factor of activated T cells (NFAT) were measured by reporter assay after transient transfection. RESULTS: RACE and RT-PCR identified a splice variant of FKBP8 lacking exons 3, 4, and 5 in human RPE cells. Both the full-length and the short form of FKBP8 proteins showed mitochondrial distribution and directly interacted with Bcl-XL. Overexpression of FKBP8 caused increased sensitivity to apoptosis induced by thapsigargin. The transcriptional activity of NFAT was not affected by FKBP8. CONCLUSIONS: FKBP8 and its novel splice variant are Bcl-XL interacting proteins and regulate the apoptotic signaling pathways in the RPE. PMID- 18385098 TI - Accommodation-related changes in monochromatic aberrations of the human eye as a function of age. AB - PURPOSE: To investigate the relationship between accommodation and the optical aberrations of the whole human eye, as a function of age. METHODS: Sixty healthy subjects with spherical ametropia in the range +/-3 D, astigmatism less than 1 D, corrected visual acuity of 20/18 or better, and normal findings in an ophthalmic examination were enrolled. Subjects were divided into four groups, with age ranges of 19 to 29, 30 to 39, 40 to 49, and 50 to 60 years. Monochromatic optical aberrations and pupil size were measured with a Hartmann-Shack wavefront sensor under monocular viewing conditions, without pharmacological dilation or cycloplegia. Stimulus vergences were in the range of 0 to 5 D, with an increment of 0.5 D. The change in aberration during accommodation for different groups and different pupil conditions (natural and fixed 4-mm pupil) was compared. RESULTS: Fourth-order spherical aberration (SA) became more negative with accommodation, and the rate of this change was greater in older individuals. For natural pupil conditions, there were no significant differences between age groups in the changes of the higher-order aberrations, coma, and trefoil with accommodation. However, for a 4-mm pupil, the youngest and oldest group showed significant differences in higher order RMS (root mean square) and spherical aberration compared with the other groups. High-order RMS showed a lower increase during accommodation when the pupil accommodative miosis was taken into account (natural pupil condition) than when a fixed 4-mm pupil was used. CONCLUSIONS: Aberrations change with accommodation and with age. SA changes more with accommodation do than other higher-order aberrations. SA becomes more negative with accommodation, and this change is larger in older individuals. Accommodative miosis is useful for ameliorating the increase in higher-order aberrations with accommodation. PMID- 18385099 TI - Therapeutic benefit derived from RNAi-mediated ablation of IMPDH1 transcripts in a murine model of autosomal dominant retinitis pigmentosa (RP10). AB - Mutations within the inosine 5'-monophosphate dehydrogenase 1 (IMPDH1) gene cause the RP10 form of autosomal dominant retinitis pigmentosa (adRP), an early-onset retinopathy resulting in extensive visual handicap owing to progressive death of photoreceptors. Apart from the prevalence of RP10, estimated to account for 5-10% of cases of adRP in United States and Europe, two observations render this form of RP an attractive target for gene therapy. First, we show that while recombinant adeno-associated viral (AAV)-mediated expression of mutant human IMPDH1 protein in the mouse retina results in an aggressive retinopathy modelling the human counterpart, expression of a normal human IMPDH1 gene under similar conditions has no observable pathological effect on retinal function, indicating that over-expression of a therapeutic replacement gene may be relatively well tolerated. Secondly, complete absence of IMPDH1 protein in mice with a targeted disruption of the gene results in relatively mild retinal dysfunction, suggesting that significant therapeutic benefit may be derived even from the suppression only component of an RNAi-based gene therapy. We show that AAV-mediated co expression in the murine retina of a mutant human IMPDH1 gene together with short hairpin RNAs (shRNA) validated in vitro and in vivo, targeting both human and mouse IMPDH1, substantially suppresses the negative pathological effects of mutant IMPDH1, at a point where, in the absence of shRNA, expression of mutant protein in the RP10 model essentially ablates all photoreceptors in transfected areas of the retina. These data strongly suggest that an RNAi-mediated approach to therapy for RP10 holds considerable promise for human subjects. PMID- 18385100 TI - Striatal and nigral pathology in a lentiviral rat model of Machado-Joseph disease. AB - Machado-Joseph disease (MJD) is a fatal, dominant neurodegenerative disorder. MJD results from polyglutamine repeat expansion in the MJD-1 gene, conferring a toxic gain of function to the ataxin-3 protein. In this study, we aimed at overexpressing ataxin-3 in the rat brain using lentiviral vectors (LV), to generate an in vivo MJD genetic model and, to study the disorder in defined brain regions: substantia nigra, an area affected in MJD, cortex and striatum, regions not previously reported to be affected in MJD. LV encoding mutant or wild-type human ataxin-3 was injected in the brain of adult rats and the animals were tested for behavioral deficits and neuropathological abnormalities. Striatal pathology was confirmed in transgenic mice and human tissue. In substantia nigra, unilateral overexpression of mutant ataxin-3 led to: apomorphine-induced turning behavior; formation of ubiquitinated ataxin-3 aggregates; alpha-synuclein immunoreactivity; and loss of dopaminergic markers (TH and VMAT2). No neuropathological changes were observed upon wild-type ataxin-3 overexpression. Mutant ataxin-3 expression in striatum and cortex, resulted in accumulation of misfolded ataxin-3, and within striatum, loss of neuronal markers. Striatal pathology was confirmed by observation in MJD transgenic mice of ataxin-3 aggregates and substantial reduction of DARPP-32 immunoreactivity and, in human striata, by ataxin-3 inclusions, immunoreactive for ubiquitin and alpha synuclein. This study demonstrates the use of LV encoding mutant ataxin-3 to produce a model of MJD and brings evidence of striatal pathology, suggesting that this region may contribute to dystonia and chorea observed in some MJD patients and may represent a target for therapies. PMID- 18385103 TI - The 2008 Thomas Hunt Morgan Medal. Michael Ashburner. PMID- 18385101 TI - The loss of methyl-CpG binding protein 1 leads to autism-like behavioral deficits. AB - Methyl-CpG binding proteins (MBDs) are central components of DNA methylation mediated epigenetic gene regulation. Alterations of epigenetic pathways are known to be associated with several neurodevelopmental disorders, particularly autism. Our previous studies showed that the loss of Mbd1 led to reduced hippocampal neurogenesis and impaired learning in mice. However, whether MBD1 regulates the autism-related cognitive functions remains unknown. Here we show that Mbd1 mutant (Mbd1(-/-)) mice exhibit several core deficits frequently associated with autism, including reduced social interaction, learning deficits, anxiety, defective sensory motor gating, depression and abnormal brain serotonin activity. Furthermore, we find that Mbd1 can directly regulate the expression of Htr2c, one of the serotonin receptors, by binding to its promoter, and the loss of Mbd1 led to elevated expression of Htr2c. Our results, therefore, demonstrate the importance of epigenetic regulation in mammalian brain development and cognitive functions. Understanding how the loss of Mbd1 could lead to autism-like behavioral phenotypes would reveal much-needed information about the molecular pathogenesis of autism. PMID- 18385105 TI - The 2008 George W. Beadle Award. Mark Johnston. PMID- 18385104 TI - The 2008 Genetics Society of America Medal. Susan Lindquist. PMID- 18385106 TI - The 2008 Genetics Society of America Award for Excellence in Education. R. Scott Hawley. PMID- 18385107 TI - The 2008 Novitski Prize. Thomas J. Silhavy. PMID- 18385109 TI - But see KITANI (1978). PMID- 18385110 TI - Probing the relative importance of molecular oscillations in the circadian clock. AB - Circadian ( approximately 24 hr) rhythms of behavior and physiology are driven by molecular clocks that are endogenous to most organisms. The mechanisms underlying these clocks are remarkably conserved across evolution and typically consist of auto-regulatory loops in which specific proteins (clock proteins) rhythmically repress expression of their own genes. Such regulation maintains 24-hr cycles of RNA and protein expression. Despite the conservation of these mechanisms, however, questions are now being raised about the relevance of different molecular oscillations. Indeed, several studies have demonstrated that oscillations of some critical clock genes can be eliminated without loss of basic clock function. Here, we describe the multiple levels at which clock gene/protein expression and function can be rhythmically regulated-transcription, protein expression, post-translational modification, and localization-and speculate as to which aspect of this regulation is most critical. While the review is focused on Drosophila, we include some discussion of mammalian clocks to indicate the extent to which the questions concerning clock mechanisms are similar, regardless of the organism under study. PMID- 18385111 TI - Reduced mismatch repair of heteroduplexes reveals "non"-interfering crossing over in wild-type Saccharomyces cerevisiae. AB - Using small palindromes to monitor meiotic double-strand-break-repair (DSBr) events, we demonstrate that two distinct classes of crossovers occur during meiosis in wild-type yeast. We found that crossovers accompanying 5:3 segregation of a palindrome show no conventional (i.e., positive) interference, while crossovers with 6:2 or normal 4:4 segregation for the same palindrome, in the same cross, do manifest interference. Our observations support the concept of a "non"-interference class and an interference class of meiotic double-strand-break repair events, each with its own rules for mismatch repair of heteroduplexes. We further show that deletion of MSH4 reduces crossover tetrads with 6:2 or normal 4:4 segregation more than it does those with 5:3 segregation, consistent with Msh4p specifically promoting formation of crossovers in the interference class. Additionally, we present evidence that an ndj1 mutation causes a shift of noncrossovers to crossovers specifically within the "non"-interference class of DSBr events. We use these and other data in support of a model in which meiotic recombination occurs in two phases-one specializing in homolog pairing, the other in disjunction-and each producing both noncrossovers and crossovers. PMID- 18385112 TI - aubergine gene overexpression in somatic tissues of aubergine(sting) mutants interferes with the RNAi pathway of a yellow hairpin dsRNA in Drosophila melanogaster. AB - AUBERGINE (AUB) is a member of the PPD family of proteins. These proteins are implicated in RNA interference. In this article we demonstrate that the expression of the aub gene and protein increase in aub(sting) mutants. We used a genetic method to test whether aub(sting) overexpression could interfere with proper functioning of the process of RNA interference in somatic tissues of Drosophila melanogaster. This method is based on a transgenic line bearing a construct in which a fragment of the yellow (y) gene is cloned to form an inverted repeat (y-IR) under the control of the upstream activation sequence (UAS) of the yeast transcriptional activator GAL4. The UAS-y-IR transgene and the Act5C-GAL4 driver were brought together on chromosome 3 via recombination. In the resulting strain (Act5C-y-IR), transcriptional activation by GAL4 constitutively produces a dsRNA hairpin bearing cognate sequences to the yellow gene causing continuing degradation of y mRNA resulting in yellow(1) (y(1)) phenocopies. In this genetic background, the mutation of any factor involved in RNAi should repress degradation of y mRNA, restoring the wild-type phenotype. We employed this genetic approach to show that an increased amount of AUBERGINE interferes with the regular functioning of the somatic RNAi pathway. PMID- 18385113 TI - A suppressor screen in chlamydomonas identifies novel components of the retinoblastoma tumor suppressor pathway. AB - The retinoblastoma (RB) protein is a eukaryotic tumor suppressor and negative cell-cycle regulator. Chlamydomonas reinhardtii cells that lack the RB homolog MAT3 show loss of size checkpoint control and deregulated cell-cycle progression leading to the production of tiny cells. We carried out an insertional mutagenesis screen to isolate bypass suppressors of mat3 (smt mutants) that reverted the mat3 cell-size defect. Previously we reported that the loci encoding Chlamydomonas homologs of E2F and DP were frequently disrupted in this screen, indicating that the architecture of the canonical RB pathway is conserved in Chlamydomonas with MAT3/RB acting as a negative regulator upstream of E2F/DP. Here, we describe four novel smt mutants that moderately suppressed the cell-size checkpoint and cell-cycle phenotypes of mat3. As single mutants, three of the smt strains displayed no obvious phenotypes, and one had a slightly small phenotype. Strikingly, several smt double-mutant combinations synergized to cause enhanced suppression of mat3 and even to cause a large-cell phenotype that is comparable to that caused by loss of DP1. Molecular characterization of one smt mutant revealed that suppression is due to a defect in a gene encoding a putative small ubiquitin-like modifier (SUMO) peptidase. Our results reveal a complex genetic network that lies downstream of MAT3/RB and implicate protein sumoylation as an important step for cell-cycle progression in cells that are missing MAT3/RB. PMID- 18385114 TI - Lgals6, a 2-million-year-old gene in mice: a case of positive Darwinian selection and presence/absence polymorphism. AB - Duplications of genes are widely considered to be a driving force in the evolutionary process. The fate of such duplicated genes (paralogs) depends mainly on the early stages of their evolution. Therefore, the study of duplications that have already started to diverge is useful to better understand their evolution. We present here the example of a 2-million-year-old segmental duplication at the origin of the Lgals4 and Lgals6 genes in the mouse genome. We analyzed the distribution of these genes in samples from 110 wild individuals and wild-derived inbred strains belonging to eight mouse species from Mus (Coelomys) pahari to M. musculus and 28 laboratory strains. Using a maximum-likelihood method, we show that the sequence of the Lgals6 gene has evolved under the influence of strong positive selection that is likely to result in its neofunctionalization. Surprisingly, despite this selection pressure, the Lgals6 gene is present in some mouse species, but not all. Furthermore, even within the species and populations where it is present, the Lgals6 gene is never fixed. To explain this paradox, we propose different hypotheses such as balanced selection and neutral retention of ancient polymophism and we discuss this unexpected result with regard to known galectin properties and response to infections by pathogens. PMID- 18385115 TI - Evolution and genetic population structure of prickly lettuce (Lactuca serriola) and its RGC2 resistance gene cluster. AB - Genetic structure and diversity of natural populations of prickly lettuce (Lactuca serriola) were studied using AFLP markers and then compared with the diversity of the RGC2 disease resistance gene cluster. Screening of 696 accessions from 41 populations using 319 AFLP markers showed that eastern Turkish and Armenian populations were the most diverse populations and might be located in the origin and center of diversity of L. serriola. Screening 709 accessions using the microsatellite MSATE6 that is located in the coding region of most RGC2 homologs detected 366 different haplotypes. Again, the eastern Turkish and Armenian populations had the highest diversities at the RGC2 cluster. The diversities at the RGC2 cluster in different populations were significantly correlated with their genomewide diversities. There was significant variation of copy number of RGC2 homologs in different populations, ranging from 12 to 22 copies per genome. The nucleotide diversities of two conserved lineages (type II) of RGC2 genes (K and L) were not correlated with diversities calculated using the MSATE6 or AFLP data. We hypothesize that the high genomewide diversity and diversity of the RGC2 cluster in eastern Turkish and Armenian populations resulted from high abiotic and biotic stresses in the regions of origin of L. serriola. PMID- 18385116 TI - Efficient control of population structure in model organism association mapping. AB - Genomewide association mapping in model organisms such as inbred mouse strains is a promising approach for the identification of risk factors related to human diseases. However, genetic association studies in inbred model organisms are confronted by the problem of complex population structure among strains. This induces inflated false positive rates, which cannot be corrected using standard approaches applied in human association studies such as genomic control or structured association. Recent studies demonstrated that mixed models successfully correct for the genetic relatedness in association mapping in maize and Arabidopsis panel data sets. However, the currently available mixed-model methods suffer from computational inefficiency. In this article, we propose a new method, efficient mixed-model association (EMMA), which corrects for population structure and genetic relatedness in model organism association mapping. Our method takes advantage of the specific nature of the optimization problem in applying mixed models for association mapping, which allows us to substantially increase the computational speed and reliability of the results. We applied EMMA to in silico whole-genome association mapping of inbred mouse strains involving hundreds of thousands of SNPs, in addition to Arabidopsis and maize data sets. We also performed extensive simulation studies to estimate the statistical power of EMMA under various SNP effects, varying degrees of population structure, and differing numbers of multiple measurements per strain. Despite the limited power of inbred mouse association mapping due to the limited number of available inbred strains, we are able to identify significantly associated SNPs, which fall into known QTL or genes identified through previous studies while avoiding an inflation of false positives. An R package implementation and webserver of our EMMA method are publicly available. PMID- 18385117 TI - Preserved left ventricular twist and circumferential deformation, but depressed longitudinal and radial deformation in patients with diastolic heart failure. AB - AIMS: To examine myocardial deformation and rotation in patients with heart failure, and elucidate the underlying mechanisms that account for normal ejection fraction (EF) in patients with diastolic heart failure (DHF). METHODS AND RESULTS: Fifty consecutive patients presenting with congestive heart failure (age: 58 +/- 16 years) underwent simultaneous right heart catheterization and transthoracic echocardiography. Left ventricular (LV) volumes, mass, EF, meridional, and circumferential wall stress were measured in addition to haemodynamic measurements. 2-D speckle tracking was applied to measure longitudinal, radial, and circumferential strain and twist. Twist was reduced only in patients with systolic heart failure (SHF: 5 +/- 2 degrees, DHF: 13 +/- 6 degrees, control: 14 +/- 5 degrees, P < 0.001). Circumferential strain was not different between DHF (-15 +/- 5%) and control groups (-20 +/- 3%, P > 0.05), though it was significantly lower in patients with SHF (-7 +/- 3%, P < 0.05). Importantly, longitudinal (DHF:-12%, SHF: -4%, control: -19%, P < 0.001) and radial (DHF: 28 +/- 9%, SHF: 14 +/- 8%, control: 47 +/- 7%, P < 0.001) strains were significantly lower in both heart failure groups than in controls, and were depressed to a larger extent in SHF patients than in those with DHF (both P < 0.05). CONCLUSION: LV longitudinal and radial strains are reduced, but circumferential deformation and twist are normal in DHF patients. On the other hand, in patients with SHF, longitudinal, radial, and circumferential deformation, and twist are all reduced. Multivariable regression analysis suggests that preserved LV twist and circumferential strain may contribute to normal EF in patients with DHF. PMID- 18385118 TI - Paced ventricular electrogram fractionation predicts sudden cardiac death in hypertrophic cardiomyopathy. AB - AIMS: Paced electrogram fractionation analysis (PEFA) has been assessed for the prediction of sudden cardiac death (SCD) in a large-scale, prospective study of patients with hypertrophic cardiomyopathy (HCM). METHODS AND RESULTS: We determined the positive predictive value (PPV) of PEFA in relation to other risk factors for SCD and outcomes in 179 patients with HCM and no prior history of cardiac arrest. Patients were followed over a mean 4.3 years (range: 1.1-6.3 years). Thirteen patients had SCD-equivalent events: four of these patients died suddenly, three were resuscitated from ventricular fibrillation (VF), and six had implantable cardioverter-defibrillator (ICD) discharges in response to VF. PEFA identified nine of these patients and another 14 non-VF patients yielding a censored PPV of between 0.19 and 0.59 that was greater than the PPV that was the formal stopping point of the trial (0.18). Eighty per cent of patients were followed for 4 years or more. The PPV for the identification of SCD in this group was 0.38 (0.17-0.59). The use of two or more conventional markers to predict SCD identified five patients with SCD-equivalent events in the 4-year follow-up group and 42 other patients without events yielding a PPV of 0.106 (confidence limits 0.02-0.15). CONCLUSION: PEFA identifies HCM patients at risk of SCD with greater accuracy than non-invasive techniques and may have an important role in determining indications for ICD prescription. PMID- 18385119 TI - Age- and gender-specific mortality rates in childhood hypertrophic cardiomyopathy. AB - AIMS: Hypertrophic cardiomyopathy (HCM) is the commonest inherited cause of sudden cardiac death in children; current guidelines suggest HCM screening after 12-15 years of age. The study aims to establish the age range at highest risk. METHODS AND RESULTS: Cohort study from six regional centres of paediatric cardiology, including children presenting with sudden death; n = 150 (59% = male; 39% familial HCM). Age- and gender-specific mortality was calculated, and compared with rates calculated from the Swedish National Cause of Death Registry. There were 56 deaths within the cohort, 39 were sudden arrhythmia deaths, with 31 at <19 years of age. Between 9-13.9 years of age annual sudden death mortality averages 7.2%, vs. 1.7% after 16 years of age; P = 0.025, odds ratio for proportions 3.75 [95% confidence intervals (CI) 1.18-11.91], similar in both familial and idiopathic HCM. The risk for sudden death peaks earlier in girls (10 11 years), with male preponderance after the age of 15. National cause of death statistics confirm that the mortality rate from HCM is significantly higher in the 8-16 year olds (0.112 per 100,000 age-specific population) than in the 17-30 year olds (0.055 per 100,000; 95% CI 0.011-0.099). CONCLUSION: In families with HCM, children should be screened at an early age. PMID- 18385120 TI - Non-invasive diagnosis of ischaemic heart failure using 64-slice computed tomography. AB - AIMS: We evaluated the accuracy of 64-slice computed tomography (CT) to identify ischaemic aetiology of heart failure (IHF). METHODS AND RESULTS: Ninety-three consecutive patients in sinus rhythm with dilated cardiomyopathy but without suspicion of coronary artery disease (CAD) were enrolled when admitted for angiography. Accuracy of CT to detect significant stenosis (>50% lumen narrowing) was compared with quantitative coronary angiography. IHF was defined as a significant stenosis on left main or proximal left anterior descending artery or two or more vessels. Forty-three out of 1395 segments (3%) were heavily calcified and excluded. CT correctly assessed 103 of 142 (73%) significant stenosis and identified 46 of 50 (92%) patients without and 42 of 43 (98%) patients with CAD, 60 of 62 (97%) patients without and 28 of 31 (90%) patients with IHF. Overall, accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of CT for identifying CAD by segment was 96, 73, 99, 92, and 97%, respectively; by patient was 95, 98, 92, 91, and 98%, respectively; and for identifying IHF was 95, 90, 97, 93, and 95%, respectively. CONCLUSION: Non invasive 64-slice CT assessment of the extent of CAD may offer a valid alternative to angiography for the diagnosis of IHF. PMID- 18385121 TI - Oedema in acute myocardial infarction. PMID- 18385122 TI - Paroxysmal reciprocating supraventricular tachycardia in infants: electrophysiologically guided medical treatment and long-term evolution of the re entry circuit. AB - AIMS: The aim of this study is to evaluate the long-term prognosis in infants affected by paroxysmal reciprocating supraventricular tachycardia (SVT), to identify predictors of SVT disappearance, and to assess the efficacy of electrophysiologically guided drug therapy in preventing recurrences. METHODS AND RESULTS: A six step regimen of oral therapy was used in 55 infants with SVT: (i) propafenone (P); (ii) flecainide (F); (iii) flecainide plus propranolol (FP); (iv) amiodarone (A); (v) amiodarone plus propranolol (AP); (vi) amiodarone plus flecainide plus propranolol (AFP). If one step was not successful, the patient was passed on to the next treatment step and so on. Transesophageal atrial pacing (TAP) was used to evaluate treatment efficacy and the evolution of SVT at the end of the first, second, and third year. Propafenone was successful in 32.7% of the patients, F in 14.5%, FP in 23.6%, A alone in 5.4%, and AP in 18.1%; only 7.2% reached step 6. At month 12, after therapy wash out, SVT recurred spontaneously in 2 patients (3.6%) and remained inducible in 25 (45.5%). Inducibility was significantly higher in patients treated with A. At 24 months, SVT was inducible or spontaneous in 86% of the cases and at 36 months in 87%. There were no recurrences using the treatment confirmed by TAP. No further predictor of SVT inducibility was identified. CONCLUSION: Supraventricular tachycardia disappeared in approximately 50% of the patients during the first year of life and in another 20% thereafter. The necessity for A treatment is the only predictor of persistence of the re-entry circuit during the first year of life. Transesophageal atrial pacing is useful in guiding the medical treatment. PMID- 18385123 TI - Catheter ablation of paroxysmal atrial fibrillation improves cardiac function: a prospective study on the impact of atrial fibrillation ablation on left ventricular function assessed by magnetic resonance imaging. AB - AIMS: Beneficial effects of atrial fibrillation (AF) ablation have been demonstrated in patients with congestive heart failure (CHF) and significantly impaired left ventricular ejection fraction (LVEF). However, the impact of pulmonary vein isolation (PVI) on cardiac function in patients with paroxysmal AF and impaired LVEF remains under discussion. This study aimed to evaluate the impact of PVI for paroxysmal AF on cardiac function in patients with impaired LVEF using cardiac magnetic resonance imaging (CMRI). METHODS AND RESULTS: A total number of 70 patients with paroxysmal AF and episodes < or = 24 h were scanned on a 1.5-T-CMRI before and 6 months after PVI during sinus rhythm. End diastolic volume, end-systolic volume, and LVEF were determined by epicardial and endocardial measurements. Patients were categorized into two groups regarding cardiac function as assessed by CMRI: group 1 patients (n = 18) with an LVEF < 50% and patients with an LVEF > 50% (group 2, n = 52). Group 1 patients demonstrated a significant lower success rate than patients of group 2 after a follow-up of 152 +/- 40 days (50 vs. 73%, P < 0.05). Cardiac magnetic resonance imaging in group 1 patients demonstrated a significant improvement in cardiac function after AF ablation (41 +/- 6 vs. 51 +/- 12%, P = 0.004), whereas group 2 patients did not show significant differences (60 +/- 6 vs. 59 +/- 9%, P = 0.22) after a 6 months follow-up. CONCLUSION: Pulmonary vein isolation improves cardiac function in patients with paroxysmal AF and impaired LVEF. These data suggest that an impaired LV function can be partially attributed to AF with short-lasting paroxysms. PMID- 18385124 TI - Heterogeneity of the mitochondrial proteome for photosynthetic and non photosynthetic Arabidopsis metabolism. AB - Heterogeneity of the mitochondrial proteome in plants underlies fundamental differences in the roles of these organelles in different tissues. We quantitatively compared the mitochondrial proteome isolated from a non photosynthetic cell culture model with more specialized mitochondria isolated from photosynthetic shoots. Differences in intact mitochondrial respiratory rates with various substrates and activities of specific enzymes provided a backdrop of the functional variation between these mitochondrial populations. Proteomics comparisons provided a deep insight into the different steady-state abundances of specific mitochondrial proteins. Combined these data showed the elevated level of the photorespiratory apparatus and its complex interplay with glycolate, cysteine, formate, and one-carbon metabolism as well as the decrease of selected parts of the tricarboxylic acid cycle, alterations in amino acid metabolism focused on 2-oxoglutarate generation, and degradation of branched chain amino acids. Comparisons with microarray analysis of these tissue types showed a positive, mild correlation between mRNA and mitochondrial protein abundance, a tighter correlation for specific biochemical pathways, but over 78% concordance in direction between changes in protein and transcript abundance in the two tissues. Overall these results indicated that the majority of the variation in the plant mitochondrial proteome occurred in the matrix, highlighted the constitutive nature of the respiratory apparatus, and showed the differences in substrate choice and/or availability during photosynthetic and non-photosynthetic metabolism. PMID- 18385125 TI - Changing etiology of tubal pregnancy following IVF. AB - BACKGROUND: Tubal pregnancy (TP) is twice as common following IVF when compared with natural conception. This is surprising, since embryo transfer is aimed for an accurate area in the uterine cavity. We thus hypothesized that either the embryo or the Fallopian tube actively participates in a pathological process leading to implantation outside the uterine cavity. Since we recently found that E-cadherin expression is a useful marker of endometrial receptivity, we considered that it may have a role in TP following IVF. Therefore, the aim of this study was to compare E-cadherin expression and localization in tubal implantation sites from spontaneous TP and TP post-IVF. METHODS: We compared E cadherin immunohistochemistry levels on cross-sections of Fallopian tubes in 11 spontaneous (antegrade) versus 13 post-IVF (retrograde) TP. The intensity of immunoreactivity was scored in a semi-qualitative blinded manner. RESULTS: The semi-quantitative intensity score in IVF tubal samples was more than double that observed in spontaneous TP (16.9 versus 7.3, respectively, P < 0.0005). E cadherin showed the most intense immunostaining in cytotrophoblast cells of chorionic villi in ectopic TP post-IVF compared with negative or weak staining in spontaneous ectopic TP. CONCLUSIONS: E-cadherin can serve as a marker of implantation. Differential expression of this adhesion molecule in TP post-IVF, when compared with natural conception, may reflect a different mechanism of embryo implantation. Moreover, the observation that E-cadherin is mostly expressed in trophoblasts, and not in the tubal wall, suggests that the preimplantation embryo may actively participate in locating a suitable implantation site. PMID- 18385126 TI - Prenatal testing among women pregnant after assisted reproductive techniques in Denmark 1995-2000: a national cohort study. AB - BACKGROUND: Women pregnant after assisted reproductive techniques (ART) are generally older than women with spontaneously conceived pregnancies, and are consequently more likely to carry a child affected by a chromosomal disorder. Furthermore, a significantly increased rate of chromosomal abnormalities after intracytoplasmatic sperm injection (ICSI) has been reported. The aim of this study was to describe the use and results of prenatal invasive diagnostic testing in a national Danish cohort of in vitro fertilization (IVF)/ICSI pregnancies. Additionally, we examined to what extent second trimester serum screening was used. METHODS: We used a register-based cohort study including all ongoing clinical pregnancies achieved by IVF/ICSI in 1995-2000 in Denmark. Data on fertility treatment, pregnancy and pregnancy outcome together with data on cytogenic testing and the use of triple test were retrieved from national statutory registers. Data on the invasive testing rate among the general Danish population were retrieved from the same national registers. RESULTS: In this 6 year period, 8531 ART pregnancies were recorded representing an unselected national ART population (6122 IVF, 2087 ICSI and 322 'IVFICSI'). The number of prenatal invasive procedures was relatively low, 16.3%, and the uptake of second trimester serum screening was very low, 7.4%. The invasive testing rate, corrected for advanced maternal age distribution, was lower in the study population than in the general population. The rate of karyotype aberrations detected by prenatal testing was 2.7% (43/1586), whereas the overall rate of pre- and post-natally detected aberrations was 0.6% (62/9625). Chromosome aberrations were more common in the ICSI-treated group compared with the IVF-treated group [1.3% (30/2297) versus 0.5% (32/6957), P < 0.0001]. This was also the case if only prenatally diagnosed chromosome aberrations were compared [4.3% (24/556) versus 1.9% (19/975), respectively, P < 0.01]. CONCLUSIONS: ART pregnancies represent a group of high-risk pregnancies with regard to chromosomal aberrations, but nevertheless their uptake of prenatal testing was low. ICSI pregnancies compared with IVF pregnancies had a higher rate of chromosomal abnormalities, even though the average maternal age was lower. PMID- 18385127 TI - Restricted expression of the human DAZ protein in premeiotic germ cells. AB - BACKGROUND: The role of the Y chromosome-encoded Deleted in Azoospermia (DAZ) gene family in spermatogenesis remains unclear. The ability of men without the DAZ gene to produce sperm, as well as the lack of selective pressure on DAZ exon sequences during evolution, casts doubts on its functional significance. Most men have four DAZ genes encoding protein isoforms that differ significantly in size. However, published western blots showed only a single "DAZ" band, raising the possibility that not all four DAZ genes are expressed. METHODS: RT-PCR, western blotting and immunostaining were used to study the expression of the four DAZ genes and the autosomal DAZL gene in human testes and in tissue culture cells. RESULTS: RNA transcripts of all four DAZ genes were found in the testis, but at much lower levels than that of the DAZL transcripts. Expression in cultured somatic cells showed that DAZ transcripts encoding multiple DAZ repeats were translated inefficiently. No DAZ proteins could be unambiguously identified on western blots when the testicular samples from three patients without the DAZ genes were used as negative controls. Nonetheless, low levels of DAZ were detected in the cytoplasm of spermatogonia by immunostaining. CONCLUSIONS: The expression of DAZ proteins in adult human testes is restricted to the spermatogonia and suggests a premeiotic role. PMID- 18385128 TI - X-ray crystallographic studies of substrate binding to aristolochene synthase suggest a metal ion binding sequence for catalysis. AB - The universal sesquiterpene precursor, farnesyl diphosphate (FPP), is cyclized in an Mg(2+)-dependent reaction catalyzed by the tetrameric aristolochene synthase from Aspergillus terreus to form the bicyclic hydrocarbon aristolochene and a pyrophosphate anion (PP(i)) coproduct. The 2.1-A resolution crystal structure determined from crystals soaked with FPP reveals the binding of intact FPP to monomers A-C, and the binding of PP(i) and Mg(2+)(B) to monomer D. The 1.89-A resolution structure of the complex with 2-fluorofarnesyl diphosphate (2F-FPP) reveals 2F-FPP binding to all subunits of the tetramer, with Mg(2+)(B)accompanying the binding of this analogue only in monomer D. All monomers adopt open activesite conformations in these complexes, but slight structural changes in monomers C and D of each complex reflect the very initial stages of a conformational transition to the closed state. Finally, the 2.4-A resolution structure of the complex with 12,13-difluorofarnesyl diphosphate (DF FPP) reveals the binding of intact DF-FPP to monomers A-C in the open conformation and the binding of PP(i), Mg(2+)(B), and Mg(2+)(C) to monomer D in a predominantly closed conformation. Taken together, these structures provide 12 independent "snapshots" of substrate or product complexes that suggest a possible sequence for metal ion binding and conformational changes required for catalysis. PMID- 18385129 TI - Metabolism of the folate precursor p-aminobenzoate in plants: glucose ester formation and vacuolar storage. AB - Plants produce p-aminobenzoate (pABA) in chloroplasts and use it for folate synthesis in mitochondria. In plant tissues, however, pABA is known to occur predominantly as its glucose ester (pABA-Glc), and the role of this metabolite in folate synthesis has not been defined. In this study, the UDP-glucose:pABA acyl glucosyltransferase (pAGT) activity in Arabidopsis extracts was found to reside principally (95%) in one isoform with an apparent K(m) for pABA of 0.12 mm. Screening of recombinant Arabidopsis UDP-glycosyltransferases identified only three that recognized pABA. One of these (UGT75B1) exhibited a far higher k(cat)/K(m) value than the others and a far lower apparent K(m) for pABA (0.12 mm), suggesting its identity with the principal enzyme in vivo. Supporting this possibility, ablation of UGT75B1 reduced extractable pAGT activity by 95%, in vivo [(14)C]pABA glucosylation by 77%, and the endogenous pABA-Glc/pABA ratio by 9-fold. The K(eq) for the pABA esterification reaction was found to be 3 x 10( 3). Taken with literature data on the cytosolic location of pAGT activity and on cytosolic UDP-glucose/UDP ratios, this K(eq) value allowed estimation that only 4% of cytosolic pABA is esterified. That pABA-Glc predominates in planta therefore implies that it is sequestered away from the cytosol and, consistent with this possibility, vacuoles isolated from [(14)C]pABA-fed pea leaves were estimated to contain> or =88% of the [(14)C]pABA-Glc formed. In total, these data and the fact that isolated mitochondria did not take up [(3)H]pABA-Glc, suggest that the glucose ester represents a storage form of pABA that does not contribute directly to folate synthesis. PMID- 18385130 TI - Molecular mechanisms of soluble cytokine receptor generation. AB - Soluble cytokine receptors play key roles in regulating cytokine-mediated biological events by binding and modulating the activity of target ligands in either an antagonistic or agonistic fashion. This Minireview will provide an overview of the molecular mechanisms mediating the generation of soluble cytokine receptors, which include sheddase-mediated proteolytic cleavage of cell-surface receptors, generation of soluble receptors by alternative gene splicing, transcription and translation of cytokine-binding genes, and extracellular release of membrane-bound receptors within vesicles such as exosomes. PMID- 18385131 TI - SSB antagonizes RecX-RecA interaction. AB - The RecX protein of Escherichia coli inhibits the extension of RecA protein filaments on DNA, presumably by binding to and blocking the growing filament end. The direct binding of RecX protein to single-stranded DNA is weak, and previous reports suggested that direct binding to DNA did not explain the effects of RecX. We now demonstrate that elevated concentrations of SSB greatly moderate the effects of RecX protein. High concentrations of the yeast RPA protein have the same effect, suggesting that the effect is not species-specific or even specific to bacterial SSB proteins. A direct SSB-RecX interaction is thus unlikely. We suggest that SSB is blocking access to single-stranded DNA. The evident competition between RecX and SSB implies that the mechanism of RecX action may involve RecX binding to both RecA protein and to DNA. We speculate that the interaction of RecX protein and RecA may enable an enhanced DNA binding by RecX protein. The effects of SSB are increased if the SSB C terminus is removed. PMID- 18385132 TI - The specific amino acid sequence between helices 7 and 8 influences the binding specificity of human apolipoprotein A-I for high density lipoprotein (HDL) subclasses: a potential for HDL preferential generation. AB - Humans have two major high density lipoprotein (HDL) sub-fractions, HDL(2) and HDL(3), whereas mice have a monodisperse HDL profile. Epidemiological evidence has suggested that HDL(2) is more atheroprotective; however, currently there is no direct experimental evidence to support this postulate. The amino acid sequence of apoA-I is a primary determinant of HDL subclass formation. The majority of the alpha-helical repeats in human apoA-I are proline-punctuated. A notable exception is the boundary between helices 7 and 8, which is located in the transitional segment between the stable N-terminal domain and the C-terminal hydrophobic domain. In this study we ask whether the substitution of a proline containing sequence (PCS) separating other helices in human apoA-I for the non proline-containing sequence (NPCS) between helices 7 and 8 (residues 184-190) influences HDL subclass association. The human apoA-I mutant with PCS2 replacing NPCS preferentially bound to HDL(2). In contrast, the mutant where PCS3 replaced NPCS preferentially associated with HDL(3). Thus, the specific amino acid sequence between helices 7 and 8 influences HDL subclass association. The wild type and mutant proteins exhibited similar physicochemical properties except that the two mutants displayed greater lipid-associated stability versus wild-type human apoA-I. These results focus new attention on the influence of the boundary between helices 7 and 8 on the properties of apoA-I. The expression of these mutants in mice may result in the preferential generation of HDL(2) or HDL(3) and allow us to examine experimentally the anti-atherogenicity of the HDL subclasses. PMID- 18385133 TI - Altered heparan sulfate structure in mice with deleted NDST3 gene function. AB - We report the generation and analysis of mutant mice bearing a targeted disruption of the heparan sulfate (HS)-modifying enzyme GlcNAc N-deacetylase/N sulfotransferase 3 (NDST3). NDST3(-/-) mice develop normally, are fertile, and show only subtle hematological and behavioral abnormalities in agreement with only moderate HS undersulfation. Compound mutant mice made deficient in NDST2;NDST3 activities also develop normally, showing that both isoforms are not essential for development. In contrast, NDST1(-/-);NDST3(-/-) compound mutant embryos display developmental defects caused by severe HS undersulfation, demonstrating NDST3 contribution to HS synthesis in the absence of NDST1. Moreover, analysis of HS composition in dissected NDST3 mutant adult brain revealed regional changes in HS sulfation, indicating restricted NDST3 activity on nascent HS in defined wild-type tissues. Taken together, we show that NDST3 function is not essential for development or adult homeostasis despite contributing to HS synthesis in a region-specific manner and that the loss of NDST3 function is compensated for by the other NDST isoforms to a varying degree. PMID- 18385134 TI - ATP-binding cassette A1-mediated lipidation of apolipoprotein A-I occurs at the plasma membrane and not in the endocytic compartments. AB - ATP-binding cassette transporter (ABC) A1 is required for the lipidation of apolipoprotein A-I to generate high density lipoprotein (HDL). This process is proposed to occur through a retro-endocytosis pathway in which apoA-I internalizes with ABCA1 and generates HDL from the endosomal compartments before resecretion. The aim of this study was to determine the route of apoA-I endocytosis and whether endocytosis contributes to HDL biogenesis. Using confocal microscopy, we found that internalized apoA-I only transiently colocalized with transferrin, a retro-endocytosis marker. Instead, apoA-I perfectly colocalized with a bulk phase uptake marker (fluorescein isothiocyanate-dextran) and, at later time points, with LysoTracker in several cell models including macrophages, fibroblasts, and baby hamster kidney cells. ABCA1 colocalized poorly with internalized apoA-I. To determine the contribution of internalized apoA-I to HDL biogenesis, we specifically removed apoA-I from the cell surface and analyzed the fate of internalized apoA-I. We found that 23% of cell-associated apoA-I was internalized at steady state. Of internalized apoA-I, only 20% was converted to HDL, and the rest was degraded, consistent with a lysosomal destination. We also found that apoA-I was released approximately five times faster from the plasma membrane than from the intracellular compartments. From these kinetic parameters, we estimated that approximately 5.6% of apoA-I that interacts with cells is degraded and that internalized apoA-I contributes to approximately 1.4% of total HDL production. We also found that blocking endocytosis with sucrose or cytochalasin D did not decrease cholesterol efflux or HDL biogenesis. We therefore conclude that the plasma membrane is the main platform where ABCA1 mediated lipidation of apoA-I occurs. PMID- 18385135 TI - L-selectin-mediated lymphocyte-cancer cell interactions under low fluid shear conditions. AB - Cell migration in blood flow is mediated by engagement of specialized adhesion molecules that function under hemodynamic shear conditions, and many of the effectors of these adhesive interactions, such as the selectins and their ligands, are well defined. However, in contrast, our knowledge of the adhesion molecules operant under lymphatic flow conditions is incomplete. Among human malignancies, head and neck squamous cell cancer displays a marked predilection for locoregional lymph node metastasis. Based on this distinct tropism, we hypothesized that these cells express adhesion molecules that promote their binding to lymphoid tissue under lymphatic fluid shear stress. Accordingly, we investigated adhesive interactions between these and other cancer cells and the principal resident cells of lymphoid organs, lymphocytes. Parallel plate flow chamber studies under defined shear conditions, together with biochemical analyses, showed that human head and neck squamous cell cancer cells express heretofore unrecognized L-selectin ligand(s) that mediate binding to lymphocyte L selectin at conspicuously low shear stress levels of 0.07-0.08 dynes/cm(2), consistent with lymphatic flow. The binding of head and neck squamous cancer cells to L-selectin displays canonical biochemical features, such as requirements for sialylation, sulfation, and N-glycosylation, but displays a novel operational shear threshold differing from all other L-selectin ligands, including those expressed on colon cancer and leukemic cells (e.g. HCELL). These data define a novel class of L-selectin ligands and expand the scope of function for L-selectin within circulatory systems to now include a novel activity within shear stresses characteristic of lymphatic flow. PMID- 18385136 TI - Seven transmembrane-spanning receptors for free fatty acids as therapeutic targets for diabetes mellitus: pharmacological, phylogenetic, and drug discovery aspects. PMID- 18385137 TI - The folding of human active and inactive extracellular superoxide dismutases is an intracellular event. AB - Human extracellular superoxide dismutase (EC-SOD) is a tetrameric glycoprotein responsible for the removal of superoxide generated in the extracellular space. Two different folding variants of EC-SOD exist based on the disulfide bridge connectivity, resulting in enzymatically active (aEC-SOD) and inactive (iEC-SOD) subunits. As a consequence of this, the assembly of the EC-SOD tetramers produces molecules with variable activity and may represent a way to regulate the antioxidant level in the extracellular space. To determine whether the formation of these two folding variants is an intra- or extracellular event, we analyzed the biosynthesis in human embryonic kidney 293 cells expressing wild-type EC-SOD. These analyses revealed that both folding variants were present in the intra- and extracellular spaces, suggesting that the formation is an intracellular event. To further analyze the biosynthesis, we constructed mutants with the capacity to generate only aEC-SOD (C195S) or iEC-SOD (C45S). The expression of these suggested that the cellular biosynthetic machinery supported the secretion of aEC SOD but not iEC-SOD. The coexpression of these two mutants did not affect the expression pattern. This study shows that generation of the EC-SOD folding variants is an intracellular event that depends on a free cysteine residue not involved in disulfide bonding. PMID- 18385138 TI - A threonine on the active site loop controls transition state formation in Escherichia coli respiratory complex II. AB - In Escherichia coli, the complex II superfamily members succinate:ubiquinone oxidoreductase (SQR) and quinol:fumarate reductase (QFR) participate in aerobic and anaerobic respiration, respectively. Complex II enzymes catalyze succinate and fumarate interconversion at the interface of two domains of the soluble flavoprotein subunit, the FAD binding domain and the capping domain. An 11-amino acid loop in the capping domain (Thr-A234 to Thr-A244 in quinol:fumarate reductase) begins at the interdomain hinge and covers the active site. Amino acids of this loop interact with both the substrate and a proton shuttle, potentially coordinating substrate binding and the proton shuttle protonation state. To assess the loop's role in catalysis, two threonine residues were mutated to alanine: QFR Thr-A244 (act-T; Thr-A254 in SQR), which hydrogen-bonds to the substrate at the active site, and QFR Thr-A234 (hinge-T; Thr-A244 in SQR), which is located at the hinge and hydrogen-bonds the proton shuttle. Both mutations impair catalysis and decrease substrate binding. The crystal structure of the hinge-T mutation reveals a reorientation between the FAD-binding and capping domains that accompanies proton shuttle alteration. Taken together, hydrogen bonding from act-T to substrate may coordinate with interdomain motions to twist the double bond of fumarate and introduce the strain important for attaining the transition state. PMID- 18385139 TI - Peroxisome proliferator-activated receptor-gamma coactivator-1alpha activation of CYP7A1 during food restriction and diabetes is still inhibited by small heterodimer partner. AB - Cholesterol 7alpha-hydroxylase (CYP7A1) catalyzes the rate-limiting step in the classic pathway of hepatic bile acid biosynthesis from cholesterol. During fasting and in type I diabetes, elevated levels of peroxisome proliferator activated receptor gamma-coactivator-1alpha (PGC-1alpha) induce expression of the Cyp7A1 gene and overexpression of PGC-1alpha in hepatoma cells stimulates bile acid synthesis. Using Ad-PGC-1alpha-RNA interference to induce acute disruption of PGC-1alpha in mice, here we show that PGC-1alpha is necessary for fasting mediated induction of CYP7A1. Co-immunoprecipitation and promoter activation studies reveal that the induction of CYP7A1 is mediated by direct interaction between PGC-1alpha and the AF2 domain of liver receptor homolog-1 (LRH-1). In contrast, the very similar PGC-1beta could not substitute for PGC-1alpha. We also show that transactivation of PGC-1alpha and LRH-1 is repressed by the small heterodimer partner (SHP). Treatment of mice with GW4064, a synthetic agonist for farnesoid X receptor, induced SHP expression and decreased both the recruitment of PGC-1alpha to the Cyp7A1 promoter and the fasting-induced expression of CYP7A1 mRNA. These data suggest that PGC-1alpha is an important co-activator for LRH-1 and that SHP targets the interaction between LRH-1 and PGC-1alpha to inhibit CYP7A1 expression. Overall, these studies provide further evidence for the important role of PGC-1alpha in bile acid homeostasis and suggest that pharmacological targeting of farnesoid X receptor in vivo can be used to reverse the increase in CYP7A1 associated with adverse metabolic conditions. PMID- 18385140 TI - Dual specificity phosphatases 18 and 21 target to opposing sides of the mitochondrial inner membrane. AB - Although large-scale approaches have identified numerous mitochondrial phosphoproteins, little is known about the mitochondrial kinases and phosphatases that regulate these phosphoproteins. Here, we identify two members of the atypical dual specificity phosphatases (DSP), DSP18 and DSP21, that are localized in mitochondria. Although DSP18 is widely expressed in several mammalian tissues, DSP21 is selectively expressed in the testes. We demonstrate that DSP18 and DSP21 are targeted to mitochondria by cryptic internal localization signals. Subfractionation of mitochondria demonstrated that DSP18 is located in the intermembrane space as a peripheral membrane protein of the inner membrane. In contrast, subfractionation of rat testis mitochondria revealed DSP21 is localized to the matrix as a peripheral membrane protein of the inner membrane. Moreover, we demonstrate that a previously reported substrate for DSP18, the stress activated protein kinase, does not localize to mitochondria in several different tissues, making it an unlikely substrate for DSP18. Finally, we show that induction of apoptosis by treatment with staurosporine causes translocation of DSP18 from the intermembrane space into the cytosol similar to other apoptogenic factors such as cytochrome c. This work rigorously demonstrates the unique location of two highly similar DSPs on opposing sides of the mitochondrial inner membrane. PMID- 18385141 TI - Effect of replacing glutamic residues upon the biological activity and stability of the circular enterocin AS-48. AB - BACKGROUND: Bacteriocins are antimicrobial peptides produced by bacteria and have a relatively narrow range of activity against closely related strains. AS-48 is a circular bacteriocin produced by Enterococcus faecalis that acts against many gram-positive and some gram-negative bacteria, and could well serve as a natural food preservative and antimicrobial agent. The structure of AS-48 is a five-helix bundle in which a hypothetical plane containing the C(alpha) atoms of E4, E20, E49 and E58 segregates a patch of positively charged residues from the rest of the hydrophobic or uncharged surface residues. OBJECTIVES: The aim of this study is to investigate the significance of the four glutamic residues with regard to the potency, stability and functionality of enterocin AS-48. METHODS: Four genetically engineered variants of AS-48 were obtained by replacing each glutamic residue with alanine by site-directed mutagenesis. Each mutant peptide was purified from E. faecalis cultures. The activity of highly concentrated samples and the MIC were determined against nine bacterial strains by the spot-assay method. Structural studies were made with circular dichroism (CD) spectroscopy. RESULTS: Occasional alterations to the net charge of AS-48 did not significantly affect its activity when high concentrations of bacteriocin were used. Nevertheless, according to the MIC values, three of the four mutated peptides showed weaker activity against the majority of the gram-positive bacteria tested. CD spectroscopy showed that the derivatives were well structured, in a similar way to those of the native molecule, with no modifications in their helix content. CONCLUSIONS: The spatial location of the Glu residues rather than their negative charge played a critical role in AS-48 target-cell specificity and bactericidal activity, because the replacement of Glu with Ala modify the interactions between neighbouring residues through their side chains and the interaction to the solvent affecting the protein stability and causing variations in the activity levels against identical organisms. PMID- 18385142 TI - Dissemination of ESBL and Qnr determinants in Enterobacter cloacae in Algeria. AB - OBJECTIVES: The aim of this study is to evaluate the prevalence and diversity of extended-spectrum beta-lactamases (ESBLs) in Enterobacter cloacae clinical isolates collected from Algerian hospitals and to verify the association with qnr genes. METHODS: MICs were determined by Etest for isolates giving positive double disc synergy tests, and all isolates were screened by PCR and sequenced, respectively, for bla(TEM), bla(CTX-M), bla(SHV) and bla(VEB) genes and for qnr genes (qnrA, qnrB, qnrS), using specific primers. RESULTS: The prevalence of ESBLs was 25/141 (17.7%) with 11, 9, 4 and 1 isolates testing positive for genes encoding CTX-M-15, CTX-M-3, SHV-12 and VEB-1, respectively. Two SHV-12 producers and one CTX-M-15 producer expressed QnrS1, one isolate produced CTX-M-15 and QnrB1 and one SHV-12 producer co-expressed QnrS1 and QnrB4. qnrA was not detected in our collection, and qnr alleles were not detected in non-ESBL-producing isolates. CONCLUSIONS: SHV-12, QnrS1, QnrB1 and QnrB4 were reported for the first time in Algeria. This study also described a co-expression of qnrS1 and qnrB4 by an SHV-12 producer isolate. PMID- 18385143 TI - In vitro activity of ceftobiprole against clinical isolates of Pseudomonas aeruginosa obtained from Canadian intensive care unit (ICU) patients as part of the CAN-ICU Study. PMID- 18385144 TI - Comment on: A survey of community-associated methicillin-resistant Staphylococcus aureus in Korea. PMID- 18385145 TI - Moxifloxacin resistance is prevalent among Bacteroides and Prevotella species in Greece. AB - OBJECTIVES: Moxifloxacin is recommended in the empirical treatment of infections involving Gram-negative anaerobes. However, current European data regarding its activity against anaerobic pathogens are limited. In order to evaluate its potency, we comparatively studied the activity of moxifloxacin against recently isolated Gram-negative anaerobes. METHODS: Four hundred and ninety-five Gram negative anaerobic clinical isolates (296 Bacteroides fragilis group, 58 non fragilis Bacteroides spp. and 141 Prevotella spp.) were prospectively recovered in six Greek hospitals. Moxifloxacin MICs were determined in comparison with those of penicillin, piperacillin/tazobactam, cefoxitin, imipenem, metronidazole and clindamycin. RESULTS: Overall moxifloxacin MIC(50) and MIC(90) were 2 and 32 mg/L, respectively. Based on the current CLSI breakpoints (susceptible, < or =2 mg/L; resistant, > or =8 mg/L), almost half of the total isolates (49%) were non susceptible to moxifloxacin (32% resistant; 17% intermediate). This was more evident among the non-fragilis Bacteroides species, where 47% of the isolates were resistant and 14% intermediate to moxifloxacin. Species variation was noticed, with the highest non-susceptible rates detected among Prevotella oralis (90%), Prevotella bivia (80%), Bacteroides thetaiotaomicron (75%), Bacteroides uniformis (70%) and Bacteroides capillosus (67%) species. Among the 19 (4%) isolates that were metronidazole non-susceptible (MIC > or = 16 mg/L), only 4 (21%) were additionally non-susceptible to moxifloxacin. CONCLUSIONS: High resistance rates to moxifloxacin among Bacteroides and Prevotella spp. were recorded, exceeding those previously reported in Europe and contraindicating its use as monotherapy for infections involving Gram-negative anaerobes without prior microbiological confirmation. For empirical usage, moxifloxacin should be combined with metronidazole in order to cover for these pathogens. PMID- 18385146 TI - It is important to identify the presence of atrial fibrillation more accurately. PMID- 18385147 TI - Multiorgan failure caused by intravascular lymphoma. PMID- 18385148 TI - Assessment of oxidative stress and endothelial dysfunction in Asian Indians with type 2 diabetes mellitus with and without macroangiopathy. AB - BACKGROUND: Enhanced oxidative stress coupled with increased expression of adhesion molecules (e.g. VCAM-1, ICAM-1) and decreased nitric oxide (NO) levels are implicated in development of atheromatous vascular lesion in diabetes. The present study addresses the correlation between oxidative stress, vascular cell adhesion molecules-1 (VCAM-1), NO end products and macroangiopathic complications in type 2 diabetes mellitus (DM). DESIGN AND METHODS: The study population consisted of three groups (i) diabetic patients with macroangiopathy (Group I); (ii) diabetic patients without macroangiopathy (Group II) and (iii) healthy controls (Group III) (n = 30, each group). RESULTS: Serum malondialdehyde(MDA) concentration was significantly higher in diabetic patients as compared to controls. Group I had significantly higher malondialdehyde level as compared to Group II (P < 0.05) (5.12 +/- 1.83 micromol/l vs. 4.22 +/- 1.03 micromol/l), suggesting higher oxidative stress in patients with macroangiopathy. Significant reduction in NO end products was observed in diabetic patients compared to controls. Levels of serum NO end products levels were further reduced in patients with macroangiopathy compared to those without macroangiopathy. Group I (971.67 +/- 230.13 ng/ml) and Group II (823.55 +/- 197.74 ng/ml) had significantly higher level of sVCAM-1 compared to healthy controls (541.14 +/- 118.25 ng/ml) (P < 0.001). Also, patients with macroangiopathy had significantly higher levels of sVCAM-1 compared to those without macroangiopathy (P < 0.05). Multiple regression analysis indicated that post-prandial blood glucose, GSH and MDA were independent predictors of sVCAM-1 level (R = 0.690, P = 0.000). CONCLUSION: It can be concluded from the present study that an enhanced oxidative stress coupled with endothelial dysfunction as indicated by reduced activity of NO pathway and enhanced expression of sVCAM-1 play an important intermediary role in the pathogenesis of macrovascular complications in type 2 DM. PMID- 18385149 TI - Need for inferior vena cava filters in cancer patients: a surrogate marker for poor outcome. AB - BACKGROUND: Cancer patients have an increased incidence of venous thromboembolism (VTE). Inferior vena cava (IVC) filters are used extensively in the US, and more than 40 000 are inserted annually. The impact on survival of cancer patients receiving IVC filters has not been studied. METHODS: A retrospective study examined 206 consecutive cancer patients with VTE to compare the effects of IVC filter placement with anticoagulation (AC) therapy on overall survival (OS), as measured from the time of VTE. Patients were classified into 3 treatment groups: AC (n = 62), IVC filter (77), or combination IVC filter + AC (67). RESULTS: Treatment groups did not differ with respect to age, sex, or albumin levels. Median OS was significantly greater in patients treated with AC (13 months) compared with those treated with IVC filters (2 months) or IVC + AC (3.25 months; P < .0002). IVC patients were 1.9 times more at risk of death than AC only (hazard ratio = .528; 95% confidence interval = .374 to .745). Multivariate analysis revealed that performance status and type of thrombus were not confounders and had no effect on OS. CONCLUSION: The need for the insertion of an IVC filter projected markedly reduced survival. Patients requiring an IVC filter rather than AC as initial therapy face a 2-fold increase in risk of death. Whether or not this therapeutic procedure has a positive impact on outcome in cancer patients is uncertain. Complications resulting from thrombosis were also analyzed in this cohort. A prospective randomized trial at our institution is addressing this issue. PMID- 18385150 TI - Intrahospital correlation of the international normalized ratio. AB - Background. Monitoring of oral anticoagulant therapy (OAT) is usually accomplished by measuring prothrombin time and the international normalized ratio (INR). However, thromboplastins have different responsiveness and sensitivity to vitamin K-dependent coagulation factors depletion. Several studies have shown INR variation when low sensitive thromboplastins are used. This study compared INR variability between two laboratories using highly sensitive thromboplastins. Methods. A total of 237 plasmas were tested, half of them from patients under OAT. Samples were tested simultaneously in two laboratories: in laboratory A, a Behring Coagulation Timer instrument and a human recombinant thromboplastin (Innovin, Dade Behring) (ISI 1.01) were used. In laboratory B, a Thrombolyzer Compact (Behnk Elektronik) and a rabbit brain thromboplastin (Simplastin Excel S, Organon Teknika) with an ISI of 1.30 were used. Statistical analysis was carried out according to the method of Bland and Altman. Results. Even though high correlation coefficients were obtained when comparing both laboratories, Bland Altman analysis showed a variation of INR between laboratories ranging from -0.77 to +1.07. After logarithmic transformation of data, these values yielded a variation of the INR either 25% below or 44% above. Conclusions. These results are clearly inadequate for clinical use because such a variation would most probably induce the clinician to make a change in warfarin dose. Standardization of instruments, reagents, and controls is warranted to decrease this variation. PMID- 18385151 TI - Serum retinol concentrations in hospitalized severe protein-energy malnourished children. AB - BACKGROUND: Vitamin A deficiency (VAD) is a major nutritional problem in many developing countries. Vitamin A status has been reported to be adversely affected in protein-energy malnutrition (PEM). OBJECTIVE: To assess and compare serum retinol concentrations in hospitalized children with severe PEM and normal ones. METHODS: A prospective series of 34 malnourished and 29 normal control children, <60 months old, hospitalized at IMIP, Recife-PE were recruited for the study between August 2004 and May 2005. Serum retinol level was assayed by high performance liquid chromatography and concentrations adjusted for presence of inflammation as evidenced by levels of C-reactive protein. Blood retinol level was compared according to nutritional status (severe PEM versus normal), gender, age, maternal schooling, family income, breastfeeding practice, residence and perceived morbidity. RESULTS: The prevalence of low serum retinol (<0.70 micromol l(-1)) was 41.2% in children with severe PEM and 24.1% in normal children. Serum retinol concentrations were lower in children whose mothers had low schooling (p = 0.025) and families with low per capita income (p = 0.049), regardless of their nutritional status. Serum retinol concentrations had similar distribution between children with severe PEM and normal, when adjusted for gender (p > 0.05), age (p > 0.05), maternal schooling (p > 0.05), family income (p > 0.05), breastfeeding practice (p > 0.05) and residence (p > 0.05). However, malnourished children with diarrhoea showed lower serum retinol concentrations (p = 0.021) compared to those without diarrhoea. CONCLUSION: VAD was prevalent in hospitalized children with severe PEM and also among normal ones although in lesser magnitude. Intervention for prevention and control of deficiency of vitamin A are recommended in settings where diarrhea is endemic and there are unfavorable socio-economical conditions. PMID- 18385152 TI - TarO: a target optimisation system for structural biology. AB - TarO (http://www.compbio.dundee.ac.uk/taro) offers a single point of reference for key bioinformatics analyses relevant to selecting proteins or domains for study by structural biology techniques. The protein sequence is analysed by 17 algorithms and compared to 8 databases. TarO gathers putative homologues, including orthologues, and then obtains predictions of properties for these sequences including crystallisation propensity, protein disorder and post translational modifications. Analyses are run on a high-performance computing cluster, the results integrated, stored in a database and accessed through a web based user interface. Output is in tabulated format and in the form of an annotated multiple sequence alignment (MSA) that may be edited interactively in the program Jalview. TarO also simplifies the gathering of additional annotations via the Distributed Annotation System, both from the MSA in Jalview and through links to Dasty2. Routes to other information gateways are included, for example to relevant pages from UniProt, COG and the Conserved Domains Database. Open access to TarO is available from a guest account with private accounts for academic use available on request. Future development of TarO will include further analysis steps and integration with the Protein Information Management System (PIMS), a sister project in the BBSRC 'Structural Proteomics of Rational Targets' initiative. PMID- 18385153 TI - Mismatched dNTP incorporation by DNA polymerase beta does not proceed via globally different conformational pathways. AB - Understanding how DNA polymerases control fidelity requires elucidation of the mechanisms of matched and mismatched dNTP incorporations. Little is known about the latter because mismatched complexes do not crystallize readily. In this report, we employed small-angle X-ray scattering (SAXS) and structural modeling to probe the conformations of different intermediate states of mammalian DNA polymerase beta (Pol beta) in its wild-type and an error-prone variant, I260Q. Our structural results indicate that the mismatched ternary complex lies in between the open and the closed forms, but more closely resembles the open form for WT and the closed form for I260Q. On the basis of molecular modeling, this over-stabilization of mismatched ternary complex of I260Q is likely caused by formation of a hydrogen bonding network between the side chains of Gln(260), Tyr(296), Glu(295) and Arg(258), freeing up Asp(192) to coordinate MgdNTP. These results argue against recent reports suggesting that mismatched dNTP incorporations follow a conformational path distinctly different from that of matched dNTP incorporation, or that its conformational closing is a major contributor to fidelity. PMID- 18385154 TI - A polycomb group protein, PHF1, is involved in the response to DNA double-strand breaks in human cell. AB - DNA double-strand breaks (DSBs) represent the most toxic DNA damage arisen from endogenous and exogenous genotoxic stresses and are known to be repaired by either homologous recombination or nonhomologous end-joining processes. Although many proteins have been identified to participate in either of the processes, the whole processes still remain elusive. Polycomb group (PcG) proteins are epigenetic chromatin modifiers involved in gene silencing, cancer development and the maintenance of embryonic and adult stem cells. By screening proteins responding to DNA damage using laser micro-irradiation, we found that PHF1, a human homolog of Drosophila polycomb-like, Pcl, protein, was recruited to DSBs immediately after irradiation and dissociated within 10 min. The accumulation at DSBs is Ku70/Ku80-dependent, and knockdown of PHF1 leads to X-ray sensitivity and increases the frequency of homologous recombination in HeLa cell. We found that PHF1 interacts physically with Ku70/Ku80, suggesting that PHF1 promotes nonhomologous end-joining processes. Furthermore, we found that PHF1 interacts with a number of proteins involved in DNA damage responses, RAD50, SMC1, DHX9 and p53, further suggesting that PHF1, besides the function in PcG, is involved in genome maintenance processes. PMID- 18385155 TI - A study of the relationships between oligonucleotide properties and hybridization signal intensities from NimbleGen microarray datasets. AB - Well-defined relationships between oligonucleotide properties and hybridization signal intensities (HSI) can aid chip design, data normalization and true biological knowledge discovery. We clarify these relationships using the data from two microarray experiments containing over three million probes from 48 high density chips. We find that melting temperature (T(m)) has the most significant effect on HSI while length for the long oligonucleotides studied has very little effect. Analysis of positional effect using a linear model provides evidence that the protruding ends of probes contribute more than tethered ends to HSI, which is further validated by specifically designed match fragment sliding and extension experiments. The impact of sequence similarity (SeqS) on HSI is not significant in comparison with other oligonucleotide properties. Using regression and regression tree analysis, we prioritize these oligonucleotide properties based on their effects on HSI. The implications of our discoveries for the design of unbiased oligonucleotides are discussed. We propose that isothermal probes designed by varying the length is a viable strategy to reduce sequence bias, though imposing selection constraints on other oligonucleotide properties is also essential. PMID- 18385156 TI - Differential stabilization of reaction intermediates: specificity checkpoints for M.EcoRI revealed by transient fluorescence and fluorescence lifetime studies. AB - M.EcoRI, a bacterial sequence-specific S-adenosyl-L-methionine-dependent DNA methyltransferase, relies on a complex conformational mechanism to achieve its remarkable specificity, including DNA bending, base flipping and intercalation into the DNA. Using transient fluorescence and fluorescence lifetime studies with cognate and noncognate DNA, we have characterized several reaction intermediates involving the WT enzyme. Similar studies with a bending-impaired, enhanced specificity M.EcoRI mutant show minimal differences with the cognate DNA, but significant differences with noncognate DNA. These results provide a plausible explanation of the way in which destabilization of reaction intermediates can lead to changes in substrate specificity. PMID- 18385158 TI - Tolerance for 8-oxoguanine but not thymine glycol in alignment-based gap filling of partially complementary double-strand break ends by DNA polymerase lambda in human nuclear extracts. AB - Ionizing radiation induces various clustered DNA lesions, including double-strand breaks (DSBs) accompanied by nearby oxidative base damage. Previous work showed that, in HeLa nuclear extracts, DSBs with partially complementary 3' overhangs and a one-base gap in each strand are accurately rejoined, with the gaps being filled by DNA polymerase lambda. To determine the possible effect of oxidative base damage on this process, plasmid substrates were constructed containing overhangs with 8-oxoguanine or thymine glycol in base-pairing positions of 3-base (-ACG or -GTA) 3' overhangs. In this context, 8-oxoguanine was well tolerated by the end-joining machinery when present at one end of the break, but not when present at both ends. Thymine glycol was less well tolerated than 8-oxoguanine, reducing gap filling and accurate rejoining by at least 10-fold. The results suggest that complex DSBs can be accurately rejoined despite the presence of accompanying base damage, but that nonplanar bases constitute a major barrier to this process and promote error-prone joining. A chimeric DNA polymerase, in which the catalytic domain of polymerase lambda was replaced with that of polymerase beta, could not substitute for polymerase lambda in these assays, suggesting that this domain is specifically adapted for gap filling on aligned DSB ends. PMID- 18385157 TI - Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box. AB - In Saccharomyces cerevisiae, multiple approaches have arrived at a consensus TATA box sequence of TATA(T/A)A(A/T)(A/G). TATA-binding protein (TBP) affinity alone does not determine TATA box function. To discover how a minimal set of factors required for basal and activated transcription contributed to the sequence requirements for a functional TATA box, we performed transcription reactions using highly purified proteins and CYC1 promoter TATA box mutants. The TATA box consensus sequence is a good predictor of promoter activity. However, several nonconsensus sequences are almost fully functional, indicating that mechanistic requirements are not the only selective pressure on the TATA box. We also found that the effect of a mutation at a certain position is often dependent on other bases within a particular TATA box. Although activators and coactivators strongly influence TBP recruitment and stability at promoters, neither Mediator, the activator Gal4-V16, nor TFIID specifically compensate for the low transcription levels of the weak TATA boxes. The addition of Mediator to purified transcription reactions did, however, increase the functional selectivity for certain consensus TATA sequences. Transcription in whole-cell extracts or in vivo with these TATA box mutants indicated that factors, other than those in our purified system, may help initiate transcription from weak TATA boxes. PMID- 18385159 TI - Pyrrole-imidazole hairpin polyamides with high affinity at 5'-CGCG-3' DNA sequence; influence of cytosine methylation on binding. AB - To investigate the binding of 5'-CpG-3' sequences by small molecules, two pyrrole (Py)-imidazole (Im) hairpin polyamides, PyImPyIm-gamma-PyImPyIm-beta-Dp (1) and PyIm-beta-Im-gamma-PyIm-beta-Im-beta-Dp (2), which recognize the sequence 5'-CGCG 3', were synthesized. The binding affinities of the 5'-CGCG-3' sequence to the Py Im hairpin polyamides were measured by surface plasmon resonance (SPR) analysis. SPR data revealed that dissociation equilibrium constants (K(d)) of polyamides 1 and 2 were 1.1 (+/- 0.3) x 10(-6) M and 1.7 (+/- 0.4) x 10(-8) M, respectively. Polyamide 2 possesses great binding affinity for this sequence, 65-fold higher than polyamide 1. Moreover, when all cytosines in 5'-CpGpCpG-3' were replaced with 5-methylcytosines ((m)Cs), the K(d) value of polyamide 2 increased to 5.8 (+/- 0.7) x 10(-9) (M), which indicated about 3-fold higher binding than the unmethylated 5'-CGCG-3' sequence. These results suggest that polyamide 2 would be suitable to target CpG-rich sequences in the genome. PMID- 18385161 TI - Doping in competition or doping in sport? AB - INTRODUCTION: Since ancient times, competitive athletes have been familiar with the use of ergogenic aids and they will probably continue to use unfair and harmful substances in future, because their inclination to victory, along with the mirage of glory and money, will probably overcome health and legal risks. SOURCES OF DATA: We searched PubMed using the term doping over the period 1990 to the present day. We also included non-English journals. AREAS OF AGREEMENT: By literature searching, it emerges that the phenomenon of doping is complex and multifaceted. It involves a number of causes and factors that do not originate solely in the athletic field, making universality its main feature. It is in fact observed in all ages and levels of competition, and it concerns all sports, even the most unpredictable. AREAS OF CONTROVERSY: The high number of athletes testing positive for anti-doping controls attests that the current strategy might be analytically adequate to unmask most (but not all) doping practices, but it is probably ineffective to prevent athletes to dope and modify this upsetting trend. Growing points As doping parallels the use of medications, food supplements, alcohol and social drugs, a reinforced preventive policy is advisable. EMERGING AREAS FOR DEVELOPING RESEARCH: The current anti-doping policy should be replaced with a more efficient and practical strategy to identify and monitor abnormal and harmful deviations of the biochemical and haematological profiles. PMID- 18385160 TI - Apoptotic signals induce specific degradation of ribosomal RNA in yeast. AB - Organisms exposed to reactive oxygen species, generated endogenously during respiration or by environmental conditions, undergo oxidative stress. Stress response can either repair the damage or activate one of the programmed cell death (PCD) mechanisms, for example apoptosis, and finally end in cell death. One striking characteristic, which accompanies apoptosis in both vertebrates and yeast, is a fragmentation of cellular DNA and mammalian apoptosis is often associated with degradation of different RNAs. We show that in yeast exposed to stimuli known to induce apoptosis, such as hydrogen peroxide, acetic acid, hyperosmotic stress and ageing, two large subunit ribosomal RNAs, 25S and 5.8S, became extensively degraded with accumulation of specific intermediates that differ slightly depending on cell death conditions. This process is most likely endonucleolytic, is correlated with stress response, and depends on the mitochondrial respiratory status: rRNA is less susceptible to degradation in respiring cells with functional defence against oxidative stress. In addition, RNA fragmentation is independent of two yeast apoptotic factors, metacaspase Yca1 and apoptosis-inducing factor Aif1, but it relies on the apoptotic chromatin condensation induced by histone H2B modifications. These data describe a novel phenotype for certain stress- and ageing-related PCD pathways in yeast. PMID- 18385163 TI - Drought stress alters water relations and expression of PIP-type aquaporin genes in Nicotiana tabacum plants. AB - Plasma membrane intrinsic proteins (PIPs), a type of aquaporins, mediate water transport in many plant species. In this study, we investigated the relationship between the functions of PIP-type water channels and water relations of tobacco plants (Nicotiana tabacum cv. Samsun) under drought stress. Drought stress treatments have led to reductions in the stomatal conductance, transpiration, water potential and turgor pressure in leaves, and also the sap flow rate and osmotic hydraulic conductance in roots. In contrast, leaf osmotic pressure was increased in response to drought stress. Interestingly, the accumulation of NtPIP1;1 and NtPIP2;1 transcripts was significantly decreased, but only that of the NtAQP1 transcript was increased under drought stress. Functional analysis using Xenopus laevis oocytes revealed that NtPIP2;1 shows marked water transport activity, but the activities of NtAQP1 and NtPIP1;1 are weak or almost negligible, respectively, when expressed alone. However, co-expression of NtPIP1;1 with NtPIP2;1 significantly enhanced water transport activity compared with that of NtPIP1;1- or NtPIP2;1-expressing oocytes, suggesting that these two aquaporins may function as a water channel, forming a heterotetramer. Heteromerization of NtPIP1;1 and NtPIP2;1 was also suggested by co-expression analyses of NtPIP1;1-GFP (green fluorescent protein) and NtPIP2;1 in Xenopus oocytes. Re-watering treatments recovered water relation parameters and the accumulation of the three NtPIP transcripts to levels similar to control conditions. These results suggest that NtPIP1;1 and NtPIP2;1 play an important role in water transport in roots, and that expression of NtPIP1;1 and NtPIP2;1 is down-regulated in order to reduce osmotic hydraulic conductance in the roots of tobacco plants under drought stress. PMID- 18385162 TI - Brief assessment of parents' attitudes toward testing minor children for hereditary breast/ovarian cancer genes: development and validation of the Pediatric BRCA1/2 Testing Attitudes Scale (P-TAS). AB - OBJECTIVE: Predictive genetic testing for hereditary breast/ovarian cancer risk (BRCA1/2 testing) is not recommended for minor children due to its lack of immediate medical benefit and potential psychological risk. Yet, tested mothers are often interested in learning about their children's cancer risks via pediatric BRCA1/2 testing, raising a host of bioethical concerns. However, no reliable or valid tool exists to formally gauge parents' interest in such testing. The aim of this study was to develop and evaluate a new measure for use in genetic research and consultation, known as the Pediatric BRCA1/2 Testing Attitudes Scale (P-TAS). METHODS: After pretest genetic counseling and provision of a blood sample for BRCA1/2 testing, the P-TAS was administered to 187 mothers of children between 8- and 21-years-old. The measure was also given to 96 of the mothers' nontested co-parents. Analyses of the factor structure and psychometric properties of the measure were performed in mothers and confirmed in their co parents. RESULTS: The two factors of the P-TAS, labeled Attitudes and Beliefs (Factor 1) and Decision Making and Communication (Factor 2), accounted for 62.9% of the variance and were reliable (Cronbach's coefficient alphas =.70 and .90, respectively); the structure and properties were largely confirmed among co parents. Validity was indicated through its convergence with related constructs. CONCLUSIONS: This new tool may be integrated into genetic counseling research to better assess parents' attitudes and interests in pediatric BRCA1/2 testing. Such information may help guide ongoing discussions about the appropriateness of testing in adolescent or young adult children. PMID- 18385164 TI - ACTIN-RELATED PROTEIN8 encodes an F-box protein localized to the nucleolus in Arabidopsis. AB - Arabidopsis encodes six nuclear actin-related proteins (ARPs), among them ARP8 is unique in having an F-box domain and an actin homology domain. Analysis of the ARP8 promoter-beta-glucuronidase (GUS) fusion suggests that ARP8 is ubiquitously expressed in all organs and cell types. Immunocytochemical analysis with ARP8 specific monoclonal antibodies revealed that ARP8 protein is localized to the nucleolus in interphase cells and dispersed in the cytoplasm in mitotic cells. The cell cycle-dependent subcellular patterns of distribution of ARP8 are conserved in other members of Brassicaceae. Our findings provide the first insight into the possible contributions of plant ARP8 to nucleolar functions. PMID- 18385165 TI - Tomato Rab11a characterization evidenced a difference between SYP121-dependent and SYP122-dependent exocytosis. AB - The regulatory functions of Rab proteins in membrane trafficking lie in their ability to perform as molecular switches that oscillate between a GTP- and a GDP bound conformation. The role of tomato LeRab11a in secretion was analyzed in tobacco protoplasts. Green fluorescent protein (GFP)/red fluorescent protein (RFP)-tagged LeRab11a was localized at the trans-Golgi network (TGN) in vivo. Two serines in the GTP-binding site of the protein were mutagenized, giving rise to the three mutants Rab11S22N, Rab11S27N and Rab11S22/27N. The double mutation reduced secretion of a marker protein, secRGUS (secreted rat beta-glucuronidase), by half, whereas each of the single mutations alone had a much smaller effect, showing that both serines have to be mutated to obtain a dominant negative effect on LeRab11a function. The dominant negative mutant was used to determine whether Rab11 is involved in the pathway(s) regulated by the plasma membrane syntaxins SYP121 and SYP122. Co-expression of either of these GFP-tagged syntaxins with the dominant negative Rab11S22/27N mutant led to the appearance of endosomes, but co expression of GFP-tagged SYP122 also labeled the endoplasmic reticulum and dotted structures. However, co-expression of Rab11S22/27N with SYP121 dominant negative mutants decreased secretion of secRGUS further compared with the expression of Rab11S22/27N alone, whereas co-expression of Rab11S22/27N with SYP122 had no synergistic effect. With the same essay, the difference between SYP121- and SYP122-dependent secretion was then evidenced. The results suggest that Rab11 regulates anterograde transport from the TGN to the plasma membrane and strongly implicate SYP122, rather than SYP121. The differential effect of LeRab11a supports the possibility that SYP121 and SYP122 drive independent secretory events. PMID- 18385166 TI - Clinical predictors of pulmonary hypertension in sarcoidosis. AB - Pulmonary hypertension (PH) adversely affects outcome in sarcoidosis and is an important predictor of mortality in these patients. Early and accurate diagnosis of this complication may improve outcome. The current authors hypothesised that integration of 6-min walk test (6MWT) as part of the evaluation leads to an earlier diagnosis of this complication. A total of 162 patients with sarcoidosis underwent 6MWT. Demographic and pulmonary function results were analysed. Patients were further assessed by echocardiography and right heart catheterisation when clinically indicated. Patients with sarcoidosis-associated PH had significantly decreased results on pulmonary function testing. They also walked shorter distances and desaturated to lower levels on 6MWT. On logistic regression analyses, significant predictors of PH were oxygen saturation <90% on 6MWT (odds ratio (OR) 12.1, 95% confidence interval (CI) 3.66-19.73) and diffusing capacity of the lung for carbon monoxide <60% predicted (OR 7.3, 95% CI 1.98-24.82). Moreover, by combining the results of oxygen saturation at 6 min with those of echocardiography, the ability to correctly predict the presence of PH by right heart catheterisation was improved. Patients with diffusing capacity of the lung for carbon monoxide <60% predicted and oxygen desaturation <90% on 6 min walk test have a high likelihood of pulmonary hypertension and should undergo further evaluation for the presence of this disorder. PMID- 18385167 TI - Nonsteroidal anti-inflammatory drugs upregulate function of wild-type and mutant CFTR. AB - Small-scale clinical trials show that treatment of cystic fibrosis (CF) patients with ibuprofen, a nonsteroidal anti-inflammatory drug, improves the symptoms of CF and slows down the decline of lung function. Paradoxically, ibuprofen inhibits ligand-stimulated CF transmembrance conductance regulator (CFTR) activity. The aim of the present study was to investigate the effects of ibuprofen on CFTR function under different conditions. Patch-clamp recordings were performed in two lines of human airway epithelial cells: IB3-8-3-7 cells, which express wild-type CFTR; and IB3-1 cells, which express the variant CFTR with deletion of phenylalanine 580 (DeltaF580CFTR). Addition of ibuprofen to the extracellular solution caused a rapid inhibition of CFTR activity in IB3-8-3-7 cells in the presence of a high intracellular concentration of cAMP, whereas ibuprofen enhanced the CFTR conductance at low levels of cAMP. Introducing ibuprofen into the interior of cells occluded the enhancing effect of ibuprofen. Notably, the variant CFTR-mediated conductance was detected in IB3-1 cells treated with myoinositol and was enhanced by ibuprofen at endogenous levels of cAMP. In summary, nonsteroidal anti-inflammatory drugs increase the function of both wild type cystic fibrosis transmembrane conductance regulator and the phenylalanine 580 deletion in cultured human airway epithelial cells at endogenous levels of cAMP. PMID- 18385168 TI - Health-related quality of life in COPD patients with chronic respiratory failure. AB - The Maugeri Respiratory Failure (MRF-28) and Severe Respiratory Insufficiency (SRI) questionnaires were recently developed to assess health-related quality of life (HRQoL) in patients with chronic respiratory failure, although not exclusively in chronic obstructive pulmonary disease (COPD) patients. The aim of the present study was to investigate whether the MRF-28 and SRI are reliable and valid HRQoL questionnaires in COPD patients with chronic hypercapnic respiratory failure (CHRF). In total, 72 COPD patients with CHRF underwent pulmonary function and exercise testing, and completed the MRF-28, the SRI, the Chronic Respiratory Questionnaire (CRQ), the Hospital Anxiety and Depression Scale, the Groningen Activity and Restriction Scale and two dyspnoea indexes. Physical domain scores of the questionnaires correlated with exercise tolerance, dyspnoea and daily activities, while psychological domains correlated strongly with anxiety and depression. Anxiety scores accounted for 51 and 56% of the total explained variance in total CRQ and SRI scores, respectively. The emphasis of the MRF-28 was restrictions in activities of daily living (52% of total variance). In conclusion, the present study showed that the Maugeri Respiratory Failure and Severe Respiratory Insufficiency questionnaires were reliable and valid questionnaires in chronic obstructive pulmonary disease patients with chronic hypercapnic respiratory failure. While the emphasis in the Maugeri Respiratory Failure questionnaire is on activities of daily living, the Severe Respiratory Insufficiency questionnaire, like the Chronic Respiratory Questionnaire, is more related to anxiety and depression. PMID- 18385169 TI - TNFA -308G>A in two international population-based cohorts and risk of asthma. AB - Genetic association studies have related the tumour necrosis factor-alpha gene (TNFA) guanine to adenine substitution of nucleotide -308 (-308G>A) polymorphism to increased risk of asthma, but results are inconsistent. The aim of the present study was to test whether two single-nucleotide polymorphisms, of TNFA and of the lymphotoxin-alpha gene (LTA), are associated with asthma, bronchial hyperresponsiveness and atopy in adults, by combining the results of two large population-based multicentric studies and conducting a meta-analysis of previously published studies. The European Community Respiratory Health Survey (ECRHS) and Swiss Cohort Study on Air Pollution and Lung and Heart Diseases in Adults (SAPALDIA) used comparable protocols, including questionnaires for respiratory symptoms and measures of lung function and atopy. DNA samples from 11,136 participants were genotyped at TNFA -308 and LTA 252. Logistic regression employing fixed and random effects models and nonparametric techniques were used. The prevalence of asthma was 6%. The TNFA -308G>A polymorphism was associated with increased asthma prevalence and with bronchial hyperresponsiveness. No consistent association was found for atopy. The LTA 252A>G polymorphism was not associated with any of the outcomes. A meta-analysis of 17 studies showed an increased asthma risk for the TNFA -308 adenine allele. The tumour necrosis factor-alpha gene nucleotide -308 polymorphism is associated with a moderately increased risk of asthma and bronchial hyperresponsiveness, but not with atopy. These results are supported by a meta-analysis of previously published studies. PMID- 18385170 TI - Keratinocyte growth factor protects against Clara cell injury induced by naphthalene. AB - Airway epithelial cells are exposed to environmental toxicants that result in airway injury. Naphthalene (NA) causes site-selective damage to Clara cells in mouse distal airways. N-terminally truncated recombinant human keratinocyte growth factor (DeltaN23-KGF) protects against acute lung injury. The present study investigated whether or not DeltaN23-KGF protects against NA-induced acute Clara cell damage by measuring airway responses specifically and in order to identify underlying molecular mechanisms. Mice were treated with DeltaN23-KGF or PBS 33 h prior to injection of 200 mg.kg body weight(-1) NA. Lung function was analysed by head-out body plethysmography. Distal airways isolated by microdissection were assessed for cell permeability using ethidium homodimer-1. Immunohistochemistry of Clara cell-specific protein in conjunction with a physical dissector was used to quantify Clara cell numbers. RNA was isolated from frozen airways in order to analyse gene expression using quantitative RT-PCR. DeltaN23-KGF prevented NA-induced airflow limitation and Clara cell permeability, and resulted in twice as many Clara cells compared with PBS pre-treatment. DeltaN23-KGF-pre-treated mice exhibited increased expression of proliferating cell nuclear antigen mRNA. Cytochrome P(450) isoform 2F2, which converts NA into its toxic metabolite, was reduced by approximately 50%. The present results demonstrate that pre-treatment with N-terminally truncated recombinant human keratinocyte growth factor protects against naphthalene-induced injury. This suggests that N-terminally truncated recombinant human keratinocyte growth factor exerts its beneficial effect through a decrease in the expression of cytochrome P(450) isoform 2F2. PMID- 18385171 TI - Pre-immigration screening process and pulmonary tuberculosis among Ethiopian migrants in Israel. AB - More than 75,000 displaced Ethiopians have moved to Israel. Tuberculosis (TB) incidence in Ethiopia is 44 times higher than in Israel (344 versus 8 cases per 100,000 inhabitants, respectively). The aims of the present retrospective cohort study were to evaluate the pre-immigration screening process initiated in 2001 on pulmonary TB (PTB) morbidity and to assess its cost-effectiveness. Ethiopian immigrants who were screened before departure (study group) were compared with those who were screened after arrival (comparison group). Between 1998 and 2005, 24,051 Ethiopian immigrants arrived in Israel. PTB was diagnosed in 332 (1.4%) immigrants, an incidence density of 325 patients per 100,000 person-yrs. PTB cumulative incidence was lower in the study group than in the comparison group: 711 compared with 1,746 patients per 100,000 immigrants, respectively (rate ratio 0.4). PTB was detected significantly earlier in the study group than in the comparison group: 193 versus 487 days after entry, respectively. Disease incidence declined significantly during the first 2 yrs following immigration. A 5-yr predictive model indicated that 98 individuals would be free of PTB, saving US$91,055 on annual treatment cost, due to screening. The pre-immigration screening process reduced pulmonary tuberculosis incidence in subsequent years following immigration. Pulmonary tuberculosis was diagnosed earlier in the screened group than in the comparison group and the process was found to be both cost-beneficial and cost-effective. PMID- 18385172 TI - Emphysema in young adult survivors of moderate-to-severe bronchopulmonary dysplasia. AB - Improved survival following extreme preterm birth complicated by bronchopulmonary dysplasia (BPD) is resulting in an increasing number of affected infants surviving to adulthood. The aim of the present pilot study was to describe the functional and structural pulmonary sequelae of moderate and severe BPD in a population of adult survivors. All babies were cared for at one institution (King Edward Memorial Hospital, Subiaco, Australia). Subjects born between 1980 and 1987 with birthweight <1,500 g and requiring supplementary oxygen at 36 weeks post-menstrual age were identified from a complete neonatal database and recruited prospectively. Local physicians were concurrently asked to refer suitable patients. Demographics, respiratory symptoms and examination results, pulmonary function tests and computed tomography images were acquired. In total, 21 subjects were studied. Of these, 12 were female, the median (range) age was 19 (17-33) yrs and 15 (71%) had persistent respiratory symptoms. The median (range) forced expiratory volume in one second (FEV(1)) z-score was -0.77 (-8.20-1.37), the forced expiratory flow at 25-75% of forced vital capacity was -1.81 (-6.00 0.75) and the diffusing capacity of the lung for carbon monoxide was -5.04 ( 13.17- -1.24). Computed tomography was carried out on 19 subjects and all had abnormal findings, with emphysema being the most common, present in 84% of subjects. The extent of radiological emphysema was inversely related to the FEV(1) z-score. Young adult survivors of moderate and severe bronchopulmonary dysplasia may be left with residual functional and characteristic structural pulmonary abnormalities, most notably emphysema. PMID- 18385173 TI - Embolisation of pulmonary arteriovenous malformations: no consistent effect on pulmonary artery pressure. AB - Increasing evidence supports the use of embolisation to treat pulmonary arteriovenous malformations (AVMs). Most pulmonary AVM patients have hereditary haemorrhagic telangiectasia (HHT), a condition that may be associated with pulmonary hypertension. The current authors tested whether pulmonary AVM embolisation increases pulmonary artery pressure (P(pa)) in patients without baseline severe pulmonary hypertension. P(pa) was measured at the time of pulmonary AVM embolisation in 143 individuals, 131 (92%) of whom had underlying HHT. Angiography/embolisation was not performed in four individuals with severe pulmonary hypertension, whose systemic arterial oxygen saturation exceeded levels usually associated with dyspnoea in pulmonary AVM patients. In 143 patients undergoing pulmonary AVM embolisation, P(pa) was significantly correlated with age, with the most significant increase occurring in the upper quartile (aged >58 yrs). In 43 patients with repeated measurements, there was no significant increase in P(pa) as a result of embolisation. In half, embolisation led to a fall in P(pa). The maximum rise in mean P(pa) was 8 mmHg: balloon test occlusion was performed in one of these individuals, and did not predict the subsequent rise in P(pa) following definitive embolisation of the pulmonary AVMs. In the present series of patients, which excluded those with severe pulmonary hypertension, pulmonary artery pressure was not increased significantly by pulmonary arteriovenous malformation embolisation. PMID- 18385175 TI - Slow breathing reduces sympathoexcitation in COPD. AB - Neurohumoral activation has been shown to be present in hypoxic patients with chronic obstructive pulmonary disease (COPD). The aims of the present study were to investigate whether there is sympathetic activation in COPD patients in the absence of hypoxia and whether slow breathing has an impact on sympathoexcitation and baroreflex sensitivity. Efferent muscle sympathetic nerve activity, blood pressure, cardiac frequency and respiratory movements were continuously measured in 15 COPD patients and 15 healthy control subjects. Baroreflex sensitivity was analysed by autoregressive spectral analysis and the alpha-angle method. At baseline, sympathetic nerve activity was significantly elevated in COPD patients and baroreflex sensitivity was decreased (5.0+/-0.6 versus 8.9+/-0.8 ms.mmHg( 1)). Breathing at a rate of 6 breaths.min(-1) caused sympathetic activity to drop significantly in COPD patients (from 61.3+/-4.6 to 53.0+/-4.3 bursts per 100 heartbeats) but not in control subjects (39.2+/-3.2 versus 37.5+/-3.3 bursts per 100 heartbeats). In both groups, slow breathing significantly enhanced baroreflex sensitivity. In conclusion, sympathovagal imbalance is present in normoxic chronic obstructive pulmonary disease patients. The possibility of modifying these changes by slow breathing may help to better understand and influence this systemic disease. PMID- 18385174 TI - Functional single nucleotide polymorphisms of the CCL5 gene and nonemphysematous phenotype in COPD patients. AB - It was previously reported that the gain-of-function -28 guanine allele of the promoter single nucleotide polymorphism (SNP; cytosine to guanine substitution of nucleotide -28 (-28C>G)) in the CC chemokine ligand 5 gene (CCL5) was associated with susceptibility to late-onset asthma in patients who developed asthma at age > or =40 yrs. The clinical diagnosis of chronic obstructive pulmonary disease (COPD) includes emphysema and small airway disease, and upregulation of CCL5 has been described in the airways of patients with COPD. It was hypothesised that CCL5 has a genetic impact upon the variable expression of emphysema in patients with COPD. Patients with COPD were studied (n = 267). All of the patients underwent pulmonary high-resolution computed tomography (CT), and visual scoring (CT score) was performed to determine emphysema severity. Three SNPs of CCL5 were genotyped, including -403G>A, -28C>G and 375T>C. A significant difference was found in CT score according to CCL5 genotype; the -28G allele was inversely associated with CT score. When the analysis was confined to 180 patients with bronchial reversibility of <15%, even stronger evidence for this association was noted. Functional single nucleotide polymorphisms in the CC chemokine ligand 5 gene were associated with milder emphysema. Together with previous findings, the present study may identify the CC chemokine ligand 5 gene as part of a common pathway in the pathogenesis of late-onset asthma and chronic obstructive pulmonary disease with milder emphysema. PMID- 18385176 TI - Asbestos induces doxorubicin resistance in MM98 mesothelioma cells via HIF 1alpha. AB - Human malignant mesothelioma (HMM), which is strongly related to asbestos exposure, exhibits high resistance to many anticancer drugs. Asbestos fibre deposition in the lung may cause hypoxia and iron chelation at the fibre surface. Hypoxia-inducible factor (HIF)-1alpha, which is upregulated by a decreased availability of oxygen and iron, controls the expression of membrane transporters, such as P-glycoprotein (Pgp), which actively extrude the anticancer drugs. The present study aimed to assess whether asbestos may play a role in the induction of doxorubicin resistance in HMM cells through the activation of HIF 1alpha and an increased expression of Pgp. After 24-h incubation with crocidolite asbestos or with the iron chelator dexrazoxane, or under hypoxia, HMM cells were tested for HIF-1alpha activation, Pgp expression, accumulation of doxorubicin and sensitivity to its toxic effect. Crocidolite, dexrazoxane and hypoxia caused HIF 1alpha activation, Pgp overexpression and increased resistance to doxorubicin accumulation and toxicity. These effects were prevented by the co-incubation with the cell-permeating iron salt ferric nitrilotriacetate, which caused an increase of intracellular iron bioavailability, measured as increased activity of the iron regulatory protein-1. Crocidolite, dexrazoxane and hypoxia induce doxorubicin resistance in human malignant mesothelioma cells by increasing hypoxia-inducible factor-1alpha activity, through an iron-sensitive mechanism. PMID- 18385177 TI - Are asymptomatic airway hyperresponsiveness and allergy risk factors for asthma? A longitudinal study. AB - Airway hyperresponsiveness (AHR) is a characteristic feature of asthma, but it is unclear whether asymptomatic AHR is associated with a higher risk of asthma. The present study assessed whether there is an association between asymptomatic AHR in adolescence and asthma in adulthood. The association between allergy and development of asthma was also investigated. A follow-up study of a general population cohort of adolescents was performed 14 yrs after baseline. Respiratory status was assessed at baseline in 1989 and at follow-up in 2003-2004 by a respiratory symptoms questionnaire, spirometry and histamine challenge. Allergy status was also assessed. The respiratory status of 199 subjects was assessed twice. In total, 91 (46%) subjects had the same AHR status in combination with respiratory symptoms at follow-up as at baseline. Adjusted for age, sex, allergy, family history of asthma and smoking history, having asymptomatic AHR was not significantly related to having asthma 14 yrs later (odds ratio (OR) 2.15, 95% confidence interval (CI) 0.67-6.83). For subjects with allergy at baseline, the OR for developing asthma was 4.45 (95% CI 1.46-13.54). Screening for asymptomatic airway hyperresponsiveness in adolescence does not identify subjects at risk of developing asthma. Conversely, the presence of allergy in adolescence does seem to be a risk factor for asthma development. PMID- 18385178 TI - Polymorphisms in the type IV collagen alpha3 gene and the risk of COPD. AB - A number of genome-wide linkage analyses have identified the 2q33.3-2q37.2 region as the most likely to contain the genes that contribute to the susceptibility to chronic obstructive pulmonary disease (COPD). It was hypothesised that the type IV collagen alpha3 (COL4A3) gene, which is one of the genes located in the 2q33.3 2q37.2 region, may act as a low-penetrance susceptibility gene for COPD. To test this hypothesis, the association of COL4A3 -1162T>C, IVS2+12C>A, P141L, G162E, H451R, P574L and *315C>A polymorphisms with the risk of COPD was investigated in a case-control study of 311 COPD patients and 386 controls. The presence of at least one 451R allele was associated with a significantly higher risk of COPD compared with the 451 H/H genotype (adjusted odds ratio 1.48, 95% confidence interval (1.03-2.14)). When the subjects were stratified according to age and COPD severity, the 451R allele was associated with a significantly higher risk of COPD only in younger individuals with severe COPD (3.02 (1.37-6.67)). In conclusion, these findings suggest that the type IV collagen alpha3 gene contributes to the genetic susceptibility to chronic obstructive pulmonary disease. PMID- 18385179 TI - Systemic inflammation, genetic susceptibility and lung function. AB - Local inflammation in airway diseases is well recognised, but less is known about the association between low-grade systemic inflammatory processes and lung function. The aim of the present study was to assess the association between inflammatory markers and lung function, taking into account polymorphisms in genes coding for inflammatory markers. In 134 post-myocardial infarction patients, six repeated measurements of C-reactive protein (CRP), interleukin (IL) 6 and fibrinogen in peripheral blood were assayed using high-sensitivity tests. Spirometry was conducted at baseline. Genotyping of single nucleotide polymorphisms was performed in genes coding for the inflammatory markers. CRP and IL-6 levels were negatively associated with forced expiratory volume in one second (FEV(1)), forced vital capacity (FVC) and mean forced expiratory flow between 25 and 75% of FVC (FEF(25-75%)). In the CRP gene, both the polymorphism rs1205 and the haplotype 2 showed a protective association with FEV(1) and FEF(25 75%), and, to a lesser extent, with FVC. rs1205 and haplotype 2 were both negatively associated with CRP levels in peripheral blood. Analysis with instrumental variables also showed a protective effect between these CRP gene polymorphisms and lung function. Results are very suggestive that heritability of lung function is at least partly controlled by the CRP gene. Applying a Mendelian randomisation approach, the study supports a causal association between low-grade general inflammation and airway diseases. PMID- 18385180 TI - A comparative study of quality control in diagnostic radiology. AB - The main objective of this comparative study was to evaluate the national quality assurance program for X-ray diagnostic radiology in Syrian governmental hospitals. Two periods were covered in this study, the first period was from 1986 to 1998 (52 hospitals and 149 X-ray machines were considered) and the second period from 1999 to 2005 (41 hospitals and 95 X-ray machines were considered). Most of the X-ray machines studied were within the acceptable performance, but few machines needed recalibration for some parameters. Considerable improvement of about 50% was reported in the second period. This improvement could be attributed to the establishment of an effective National Regulatory Authority in Syria in 1998 that introduced and gradually enforced the quality assurance requirement for X-ray equipment as part of the licensing process and to the relatively newer X-ray machines covered in the second period. PMID- 18385181 TI - Interventional cardiovascular procedures in Belgium: effective dose and conversion factors. AB - Effective dose (E), representing the risk of late radiation-induced effects, can be estimated by the use of conversion factors (CF), converting direct measurable quantities such as dose-area-product into E. Eight Belgian hospitals participated in the study with a total number of 318 procedures. E-values, calculated with PCXMC, were compared for the different hospitals for diagnostic and therapeutic procedures separately. E-values varied significantly depending on the hospital where the procedure was performed (P < 0.001), on filtration insertion (P < 0.001), on whether a centre is a training centre or not, the dose conscious action of the cardiologists and the complexity of the procedure (P < 0.001). Hospital-specific CF were calculated. An average CF of 0.185 mSv Gycm(-2) was obtained with a satisfactory correlation (r = 0.966, P < 0.001). The differences in CF between hospitals were due to, a large extent, the availability of additional filtration in cinegraphy mode (P < 0.001) and not to the differences in irradiation geometries in the clinical protocol of the interventional procedures. PMID- 18385182 TI - Proceedings of the 5th Conference on Protection against Radon at Home and at Work. September 9-15, 2007. Prague, Czech Republic. PMID- 18385183 TI - Clinical outcomes of progressive supranuclear palsy and multiple system atrophy. AB - Prognostic predictors have not been defined for progressive supranuclear palsy (PSP) and multiple system atrophy (MSA). Subtypes of both disorders have been proposed on the basis of early clinical features. We performed a retrospective chart review to investigate the natural history of pathologically confirmed cases of PSP and MSA. Survival data and several clinically relevant milestones, namely: frequent falling, cognitive disability, unintelligible speech, severe dysphagia, dependence on wheelchair for mobility, the use of urinary catheters and placement in residential care were determined. On the basis of early symptoms, we subdivided cases with PSP into 'Richardson's syndrome' (RS) and 'PSP parkinsonism' (PSP-P). Cases of MSA were subdivided according to the presence or absence of early autonomic failure. Sixty-nine (62.7%) of the 110 PSP cases were classified as RS and 29 (26.4%) as PSP-P. Of the 83 cases of MSA, 42 (53.2%) had autonomic failure within 2 years of disease onset. Patients with PSP had an older age of onset (P < 0.001), but similar disease duration to those with MSA. Patients with PSP reached their first clinical milestone earlier than patients with MSA (P < 0.001). Regular falls (P < 0.001), unintelligible speech (P = 0.04) and cognitive impairment (P = 0.03) also occurred earlier in PSP than in MSA. In PSP an RS phenotype, male gender, older age of onset and a short interval from disease onset to reaching the first clinical milestone were all independent predictors of shorter disease duration to death. Patients with RS also reached clinical milestones after a shorter interval from disease onset, compared to patients with PSP-P. In MSA early autonomic failure, female gender, older age of onset, a short interval from disease onset to reaching the first clinical milestone and not being admitted to residential care were independent factors predicting shorter disease duration until death. The time to the first clinical milestone is a useful prognostic predictor for survival. We confirm that RS had a less favourable course than PSP-P, and that early autonomic failure in MSA is associated with shorter survival. PMID- 18385184 TI - Distinct mechanisms of impairment in cognitive ageing and Alzheimer's disease. AB - Similar manifestations of functional decline in ageing and Alzheimer's disease obscure differences in the underlying cognitive mechanisms of impairment. We sought to examine the contributions of top-down attentional and bottom-up perceptual factors to visual self-movement processing in ageing and Alzheimer's disease. We administered a novel heading discrimination task requiring subjects to determine direction of simulated self-movement from left or right offset optic flow fields of several sizes (25 degrees, 40 degrees or 60 degrees in diameter) to 18 Alzheimer's disease subjects (mean age = 75.3, 55% female), 21 older adult control subjects (mean age = 72.4, 67% female), and 26 younger control subjects (mean age = 26.5, 63% female). We also administered computerized measures of processing speed and divided and selective attention, and psychophysical measures of visual motion perception to all subjects. Both older groups showed significant difficulty in judging the direction of virtual self-movement [F(2,194) = 40.5, P < 0.001] and optic flow stimulus size had little effect on heading discrimination for any group. Both older groups showed impairments on measures of divided [F(2,62) = 22.2, P < 0.01] and selective [F(2,62) = 63.0, P < 0.001] attention relative to the younger adult control group, while the Alzheimer's disease group showed a selective impairment in outward optic flow perception [F(2,64) = 6.3, P = 0.003] relative to both control groups. Multiple linear regression revealed distinct attentional and perceptual contributions to heading discrimination performance for the two older groups. In older adult control subjects, poorer heading discrimination was attributable to attentional deficits (R(2) adj = 0.41, P = 0.001) whereas, in Alzheimer's disease patients, it was largely attributable to deficits of visual motion perception (R(2) adj = 0.57, P < 0.001). These findings suggest that successive attentional and perceptual deficits play independent roles in the progressive functional impairments of ageing and Alzheimer's disease. We speculate that the attentional deficits that dominate in older adults may promote the development of the perceptual deficits that further constrain performance in Alzheimer's disease. PMID- 18385185 TI - Ataxia caused by amiodarone in older people. AB - Amiodarone is recommended for the cardioversion of atrial fibrillation and prevention of paroxysmal atrial fibrillation in patients with structural heart disease, coronary artery disease or left ventricular dysfunction. It has well recognised side-effects on the skin, lungs, liver, thyroid and eyes. Neurological side-effects, including ataxia and neuropathy, also occur, and may be more prevalent in older patients. These side-effects are reversible after cessation of amiodarone. Monitoring of amiodarone therapy should include assessment of the central and peripheral nervous system especially in older patients. PMID- 18385186 TI - Rate of accidental falls in institutionalised older people with and without cognitive impairment halved as a result of a staff-oriented intervention. AB - OBJECTIVE: to evaluate the impact of a staff-oriented intervention on the number of accidental falls in residents with and without cognitive impairment. DESIGN: clustered randomised controlled trial. METHODS: ten nursing wards from 7 nursing homes were randomised in a control (5 wards) and intervention (5 wards) group. The nurses from the intervention group received multi-faceted training about the occurrence of accidental falls, risk factors for falls and possible environmental modifications. For each fall they were asked to record the relevant risk factors, to keep a fall diary and to evaluate fall causes and possible preventive actions. For all residents, cognition and mobility were evaluated using a Mini-Mental State Examination (MMSE) and a Timed Up and Go Test (TUGT). Fall rates were recorded in an identical way for 6 months before and after the start of the intervention. MAIN OUTCOME MEASURES: primary outcome measure was the number of participants with at least one accidental fall requiring an intervention by a physician or a nurse during each period of recording. Secondary outcome was the number of falls for each participant during each period of recording. RESULTS: the relative risk of falling at least once in people of the intervention versus the control group adjusted for the pre-intervention results was 0.46 (95% CI: 0.26-0.79). There was no difference between residents with and without cognitive impairment or impaired mobility. In those falling at least once, the difference between the average number of falls in the two intervention arms was not significant (P = 0.10). CONCLUSION: a simple staff-oriented intervention had a substantial effect on the frequency of accidental falls. PMID- 18385187 TI - Are the take-off and landing phase dynamics of the volleyball spike jump related to patellar tendinopathy? AB - OBJECTIVE: The causal mechanism of the chronic sports injury patellar tendinopathy is not well understood. The aim of the present study was to compare ankle and knee joint dynamics during the performance of the volleyball spike jump between healthy volleyball players (n = 8) and asymptomatic volleyball players with previous patellar tendinopathy (n = 7). DESIGN: Cross-sectional. METHODS: Inverse dynamics were used to estimate ankle and knee joint dynamics. From these multiple biomechanical variables, a logistic regression was performed to estimate the probability of the presence or absence of previous patellar tendinopathy among the volleyball players studied. RESULTS: Several biomechanical variables improved the prediction of the presence or absence of previous patellar tendinopathy. For landing, ankle plantar flexion at the time of touch-down, and knee range of motion during the first part of impact, and for take-off, loading rate of the knee extensor moment during the eccentric countermovement phase of take-off were predictive. As interaction effects, the presence or absence of previous patellar tendinopathy were correctly predicted by ankle and knee range of motion during the first part of impact, by loading rate of the knee extensor moment during the eccentric phases of take-off and landing, and by knee angular velocity during the eccentric phases of take-off and landing. CONCLUSION: Smaller joint flexion during the first part of landing impact , and higher rate of knee moment development during the eccentric phases of the spike-jump landing sequence, together with higher knee angular velocities, might be risk factors in the development of patellar tendinopathy in volleyball players. PMID- 18385188 TI - Head, face and neck injury in youth rugby: incidence and risk factors. AB - OBJECTIVES: In this study, the incidence of head, neck and facial injuries in youth rugby was determined, and the associated risk factors were assessed. DESIGN: Data were extracted from a cluster randomised controlled trial of headgear with the football teams as the unit of randomisation. No effect was observed for headgear use on injury rates, and the data were pooled. SETTING: General school and club-based community competitive youth rugby in the 2002 and 2003 seasons. PARTICIPANTS: Young male rugby union football players participating in under-13, under 15, under 18 and under 21 years competitions. Eighty-two teams participated in year 1 and 87 in year 2. MAIN OUTCOME MEASURES: Injury rates for all body regions combined, head, neck and face calculated for game and missed game injuries. RESULTS: 554 head, face and neck injuries were recorded within a total of 28 902 h of rugby game exposure. Level of play and player position were related to injury risk. Younger players had the lowest rates of injury; forwards, especially the front row had the highest rate of neck injury; and inside backs had the highest rate of injuries causing the player to miss a game. Contact events, including the scrum and tackle, were the main events leading to injury. CONCLUSION: Injury prevention must focus on the tackle and scrum elements of a youth rugby game. PMID- 18385189 TI - Comparison of fat oxidation during exercise in lean and obese pubertal boys: clinical implications. AB - OBJECTIVE: To examine fat oxidation rates during exercise in lean and obese pubescent children. DESIGN: A graded leg cycle ergometry test was performed by two groups of pubescent boys (13 lean: mean (SD) age 12.0 (0.5) years, body mass index (BMI) 18.56 (1.12) kg/m(2); 17 obese: mean (SD) age 12.1 (0.1) years, BMI 26.68 (3.37) kg/m(2); p<0.001). The first step of the test was fixed at 30 W and power was gradually increased by 20 W every 3.5 min. The mean ventilatory gas measurement was obtained during the last 30 s of each step for calculation of fat oxidation rate vs exercise intensity. RESULTS: At low intensity (0-30% of peak oxygen consumption) when fat-free mass is considered, the fat oxidation rate was identical for the two groups. At higher intensities (40%, 50% and 60% of peak oxygen consumption) the fat oxidation rate was significantly higher in lean boys than in obese boys. CONCLUSION: These results confirm that obese pubertal boys have fat-free mass decreased capacities to use fat during moderate exercise. The findings suggest that obese boys need to practise physical activity at a lower intensity than healthy boys to enhance lipolysis and diminish adipose tissue and the consequences of obesity. PMID- 18385191 TI - MRI evaluation of body composition changes in wrestlers undergoing rapid weight loss. AB - PURPOSE: Changes in body composition of college wrestlers undergoing rapid weight reduction were evaluated over time using magnetic resonance imaging (MRI). METHODS: This study evaluated 12 wrestlers (male, 18-22 years of age) who participated in Japan's 2005 intercollegiate wrestling tournament. For this study, MRI (of the right femoral region and the trunk), as well as measurements of body weight, body fat percentage and body water content, were performed 1 month and 1 week prior to the weigh-in, on the day of the weigh-in, on the day of the match (after the match), and 1 week after the weigh-in. A survey of food and fluid intake was also conducted. RESULTS: Several variables were significantly lower on the day of the weigh-in than one month prior: body weight (p<0.01, -7.3% (SD 1.6%)); body fat (p<0.05, -9.3 (5.8)%); body water content (p<0.01, -5.9 (1.6)%); trunk cross-section (p<0.01, -13.2 (4.4)%), including separate measurements of trunk viscera, trunk muscle, and trunk fat; quadriceps muscle; lower subcutaneous; and food intake (p<0.01, 122 (20)). At 1 week after the match, all metrics had recovered to their levels measured 1 month before the weigh-in. Certain variables that were highly sensitive to hydration recovered more rapidly: they had reached their initial levels when measured immediately after the match. CONCLUSION: Rapid weight reduction reduced the wrestlers' cross sectional areas of muscle and fat tissues, which tended to recover through rehydration after the weigh-in. These results suggest that rapid weight reduction of wrestlers induced changes in different regions of the body. PMID- 18385192 TI - Regular Tai Chi Chuan exercise improves T cell helper function of patients with type 2 diabetes mellitus with an increase in T-bet transcription factor and IL-12 production. AB - BACKGROUND: Exercise has been shown to be beneficial in the treatment of type 2 diabetes mellitus (DM); its benefit to immune function, however, remains to be determined. OBJECTIVE: This study investigated the effect of a 12-week course of Tai Chi Chuan (TCC) exercise on T cell helper (Th) reaction in patients with type 2 DM. METHODS: A case-control study was performed in 30 pairs of patients with type 2 DM and normal age-matched adults. Fasting blood glucose, HbA1c, mediators (interleukin (IL)-12, IL-4 and transforming growth factor (TGF)beta) and transcription factors (T-bet, GATA-3 and FoxP3) of Th1/Th2/T regulatory (Treg) reaction were measured before and after a 12-week TCC exercise programme. RESULTS: Fasting glucose and HbA1c levels in the patients with type 2 DM were significantly higher than in age-matched controls before exercise. After TCC exercise, HbA1c levels in patients with type 2 DM significantly decreased (7.59 (0.32)% vs 7.16 (0.22)%; p = 0.047) and blood levels of IL-12 increased significantly (5.96 (1.10) vs 12.96 (3.07); p = 0.035). To study the molecular Th1/Th2/Treg reaction, patients with type 2 DM were found to have lower T-bet but not GATA-3 or FoxP3 expression than normal controls before TCC exercise. After the 12-week TCC exercise T-bet expression significantly increased in patients with type 2 DM. CONCLUSIONS: A 12-week TCC exercise programme decreases HbA1c levels along with an increase in the Th1 reaction. A combination of TCC with medication may provide an even better improvement in both metabolism and immunity of patients with type 2 DM. PMID- 18385194 TI - Personal best marathon performance is associated with performance in a 24-h run and not anthropometry or training volume. AB - OBJECTIVE: In this study, the influence of anthropometric and training parameters on race performance in ultra-endurance runners in a 24-h run was investigated. DESIGN: Descriptive field study. SETTING: 24-h run in Basel 2007. PARTICIPANTS: 15 male Caucasian ultra-runners (mean (SD) 46.7 (5.8 years), 71.1 (6.8 kg), 1.76 (0.07 m), body mass index 23.1 (1.84 kg/m(2))). INTERVENTIONS: None. MAIN OUTCOME MEASURES: Age, body mass, body height, length of lower limbs, skin-fold thicknesses, circumference of extremities, skeletal muscle mass, body mass, percentage of body fat, and training volume in 15 successful finishers were determined to correlate anthropometric and training parameters with race performance. RESULTS: No significant association (p>0.05) was found between the reached distance and the anthropometric properties. There was also no significant association between the reached distance with the weekly training hours, running years, the number of finished marathons and the number of finished 24-h runs. The reached distance was significantly (p<0.05) positively correlated with the personal best marathon performance (r(2) = 0.40) and the personal best 24-h run distance (r(2) = 0.58). Furthermore, the personal best marathon performance was significantly and positively correlated (p<0.01) with the best personal 24-h run distance (r(2) = 0.76). CONCLUSIONS: Anthropometry and training volume does not seem to have a major effect on race performance in a 24-h run. Instead, a fast personal best marathon time seems to be the only positive association with race performance in a 24-h run. PMID- 18385193 TI - Association between walking distance and percentiles of body mass index in older and younger men. AB - OBJECTIVE: To assess the association of weekly walking distance to body weight and waist circumference in elderly (age > or =75 years), senior (55< or = age <75 years), middle-aged (35< or = age <55 years), and younger men (18< or = age <35 years old). DESIGN: Cross-sectional analyses of baseline questionnaires from 7082 male participants of the National Walkers' Health Study. RESULTS: Standard regression analyses showed that body mass index (BMI) was inversely and significantly associated with walking distance (kg/m(2) per km/week) in elderly (slope (SE): -0.032 (0.008)), senior (-0.045 (0.005)) and middle-aged men (-0.037 (0.007)), as were their waist circumferences (-0.090 (0.025), -0.122 (0.012) and 0.091 (0.015) cm per km/week, respectively), and that these slopes remained significant when adjusted statistically for reported weekly servings of meat, fish, fruit and alcohol. However, percentile regression analyses showed that the declines in BMI per km/week walked were greater at the higher than the lower percentiles of the BMI distribution. In men > or =74 years old the decline per km walked was 4.9-fold greater among the heaviest men (that is, 90th BMI percentile; -0.076 kg/m(2) per km/week) than among the leanest men (that is, 10th BMI percentile; -0.015 kg/m(2) per km/week). The differences in slope at the 90th compared to the 10th BMI percentile were 5.4-fold among men 55-74 years old and sixfold among men 35-54 years old. Per km/week walked, the declines at the 90th percentile of waist circumference were also greater than at its 10th percentile, and intermediate for percentiles in between. Whereas standard regression analyses suggest that the average declines in BMI per km/week walked reported here are consistent with those reported previously per km/week run in male runners 35-54 years old (-0.036 (0.001) kg/m(2) per km/week) and > or =50 years old (-0.038 (0.001) kg/m(2) per km/week), percentile regression analyses showed that when adjusted to the leaner body weights of the runners the declines per km walked were between 49% and 59% less for walkers than runners. CONCLUSIONS: Declines in BMI and waist circumferences with walking distance depend upon the percentile of the BMI distribution, with the decline per km walked being significantly greater among heavier men. PMID- 18385195 TI - Epidemiology of cervical spine abnormalities in asymptomatic adult professional rugby union players using static and dynamic MRI protocols: 2002 to 2006. AB - OBJECTIVE: In this study, the prevalence of abnormalities in the cervical spine of asymptomatic professional rugby players using both static and dynamic magnetic resonance imaging (MRI) in order to improve the detection of abnormalities and prevention of related injuries was investigated. DESIGN: Prospective observational study. SETTING: French professional rugby union clubs, between 2002 and 2006. PARTICIPANTS: 206 elite male adult players. INTERVENTION: Static sagittal T2 and axial T2* fast spin echo (FSE), and dynamic sagittal single-shot FSE weighted MRI scans of the C2-C7 region were examined for the presence of abnormalities. Participants' spines were in supine neutral position for the static protocol but were allowed complete flexibility in a sagittal plane for the dynamic protocol. MAIN OUTCOME MEASUREMENTS: The medulla-to-canal ratio (MCR) was measured at every vertebral disc level for both MRI methods. When observed, anatomical abnormalities were categorised. RESULTS: Anatomical abnormalities mainly consisted of degenerative discopathy and were most frequently observed in players aged>21 years, as well as in players whose MCR was abnormally high based on medical expertise. Most MCRs that were initially assessed as intermediate with static MRI were subsequently assessed as abnormal with dynamic MRI assessment. CONCLUSIONS: Since dynamic MRI is more accurate than static techniques in examining the cervical spine, it contributes substantially to identifying the risk of spinal injuries in professional rugby players, and when used in association with clinical assessment, it can assist in preventing spinal injury. PMID- 18385196 TI - Effects of petrissage massage on fatigue and exercise performance following intensive cycle pedalling. AB - OBJECTIVE: Petrissage is assumed to influence circulation as well as interstitial drainage of both superficial and deep tissues. To study its effect it was applied between consecutive bouts of supramaximal exercise performed by the lower leg muscles. METHODS: Subjects were 11 healthy female students actively engaged in sports. Exercise bouts of ergometer cycling at loads determined individually (0.075 kp x body weight (kg)) for 5 s repeated eight times at intervals of 20 s had to be performed twice on an experimental day with 35 min intermittent bed rest. Each subject was investigated on two occasions with a minimum interval of 1 week, once without (control, CO) and once with 10 min petrissage (massage, MA) of the exercising lower leg during the bed rest phase. Effects of exercise bouts on blood lactate, muscle stiffness and perceived lower limb fatigue and their recovery before and after the second exercise bout were determined. RESULTS: For the first exercise bouts total power did not differ between MA and CO. Courses of blood lactate did not differ between MA and CO. However, recovery from measured muscle stiffness (p<0.05) and perceived lower limb fatigue (p<0.05) were more pronounced and total power during the second exercise bout was enhanced (p<0.01) in MA as compared with CO subjects. CONCLUSION: Petrissage improved cycle ergometer pedalling performance independent of blood lactate but in correlation with improved recovery from muscle stiffness and perceived lower limb fatigue. PMID- 18385197 TI - Health information styles among participants in a prostate cancer screening informed decision-making intervention. AB - The purpose of this study was to assess the usefulness of a health information styles segmentation strategy in understanding audience subgroups. We examined the health information styles of men participating in a prostate cancer screening informed decision-making (IDM) intervention and assessed intervention effects on men with distinct health information styles. We classified participants into three health information style groups based on their independence in health decision making (independent versus doctor dependent) and engagement in health enhancement (active versus passive): independent active (IA), doctor-dependent active (DDA) and passive. We developed profiles of men in these groups: IAs tended to be white and more highly educated and to have greater baseline prostate cancer knowledge; DDAs were older, less healthy and more likely to have visited a doctor in the past year and passives tended to be younger, not to have had a recent prostate-specific antigen test and to have lower self-efficacy related to communication with doctors and less positive interactions with doctors. All groups significantly increased their prostate cancer knowledge after the intervention, but passives exhibited the greatest increase in knowledge at a 6 month follow-up. The health information styles segmentation strategy used in this study offers a viable framework for segmenting audiences. PMID- 18385198 TI - Temsirolimus safety profile and management of toxic effects in patients with advanced renal cell carcinoma and poor prognostic features. AB - BACKGROUND: Temsirolimus, a novel inhibitor of mammalian target of rapamycin, has demonstrated prolonged overall survival and progression-free survival compared with interferon alfa (IFN) in patients with advanced renal cell carcinoma (RCC) and poor prognostic features. Adverse events (AEs) of any causality were previously reported, but AEs that were deemed temsirolimus related are of particular relevance for poor-risk patients and for defining mammalian target of rapamycin inhibitor-specific side-effects. PATIENTS AND METHODS: Patients with advanced RCC, no prior systemic therapy, and three or more of six poor-risk factors were randomly assigned to one of three groups: (i) IFN s.c. up to 18 MU thrice weekly, (ii) temsirolimus i.v. 25 mg weekly, or (iii) temsirolimus i.v. 15 mg weekly plus interferon s.c. 6 MU thrice weekly. RESULTS: Among 208 patients, the most common temsirolimus-related grades 3-4 AEs were anemia (13%), hyperglycemia (9%), and asthenia (8%). Grades 3-4 hypercholesterolemia (1%), hypertriglyceridemia (3%), and hypophosphatemia (4%) were also seen. Although pneumonitis occurred infrequently, vigilance for its development is needed. Guidelines for management of toxic effects are presented on the basis of available clinical experience. CONCLUSIONS: Temsirolimus-related grades 3-4 AEs were primarily metabolic in nature and easily controlled medically. In general, these did not negatively impact patient quality of life. PMID- 18385199 TI - The potential role of positron emission tomography in the detection of occult cancer in 25 patients with venous thromboembolism. PMID- 18385200 TI - Clinical impact of the methotrexate resistance-associated genes C-MYC and dihydrofolate reductase (DHFR) in high-grade osteosarcoma. AB - BACKGROUND: Aims of this study were the validation of C-MYC involvement in methotrexate (MTX) resistance and the assessment of clinical impact of C-MYC and dihydrofolate reductase (DHFR) in osteosarcoma (OS). MATERIALS AND METHODS: The involvement of C-MYC in MTX resistance was validated with an antisense approach. C-MYC and DHFR protein levels at diagnosis were assessed by immunohistochemistry on series of patients treated with either a MTX-based protocol (IOR/OS-1; 72 patients) or with a standard four-drug regimen (ISG/SSG 1; 61 patients). RESULTS: Down-regulation of C-MYC significantly decreased the MTX resistance level of OS cells, demonstrating its causal involvement in this phenomenon. In clinical samples, a worse outcome was associated with increased levels of DHFR and C-MYC at diagnosis in the IOR/OS-1 patients and of C-MYC in the ISG/SSG 1 patients. CONCLUSIONS: Meanwhile the adverse clinical impact of DHFR overexpression appeared to be closely related to the relevance of MTX in the chemotherapeutic protocol, that of C-MYC overexpression was more general and not strictly MTX related. The assessment of C-MYC and DHFR at diagnosis, together with that of other known prognostic markers, can be considered for an early identification of subgroups of OS patients with higher risk of adverse outcome. PMID- 18385201 TI - Clinical heterogeneity of extranodal NK/T-cell lymphoma, nasal type: a national survey of the Korean Cancer Study Group. AB - BACKGROUND: This national survey was undertaken to propose the classification of extranodal natural killer (NK)/T-cell lymphoma (NTCL) subtypes and to clarify a clinical heterogeneity. PATIENTS AND METHODS: Two hundred and eighty patients newly diagnosed as NTCL were enrolled from 22 Korean medical centers. Two subsets were compared: one involving the upper aerodigestive tract (UAT) and another involving the non-upper aerodigestive tract (NUAT) region, which comprises the skin, gastrointestinal tract, and liver or soft tissues. Clinical prognostic factors, survival outcomes, and independent predictors for survival were compared between each subset. RESULTS: NUAT-NTCL (59 patients) had significantly higher proportions of disseminated disease, aggressive biologic features, and unfavorable host reactions compared with UAT-NTCL (221 patients). NUAT-NTCL had shortened 5-year overall survival (OS) (22% versus 41%, P = 0.001). Ann Arbor staging, the International Prognostic Index, and the NTCL prognostic index failed to predict the OS of NUAT-NTCL, but did predict the OS in UAT-NTCL. Independent predictors for OS by multivariate analyses differed between each subset. In the NUAT subset, extranodal sites and regional nodes predicted the OS, while Ann Arbor staging, age, performance status, and lactate dehydrogenase level predicted the OS in the UAT subset. CONCLUSION: NUAT-NTCL may represent a distinctive disease entity in terms of clinical factors, independent predictors, and survival outcomes. PMID- 18385203 TI - A phase II multicenter study of oxaliplatin in combination with paclitaxel in poor prognosis patients who failed cisplatin-based chemotherapy for germ-cell tumors. AB - BACKGROUND: The aim of this study is to determine feasibility and efficacy of the combination regimen oxaliplatin and paclitaxel in patients with cisplatin (CDDP) refractory germ-cell tumors (GCT). PATIENTS AND METHODS: Patients with either a cisplatin absolute-refractory GCT defined as progressive disease (PD) during or within 1 month of CDDP administration or with a poor prognosis relapse, defined as PD between the second and the sixth month after CDDP administration, were treated with a combination of oxaliplatin (130 mg/m(2)) and paclitaxel (175 mg/m(2)) administered every 21 days. Primary end point was efficacy. RESULTS: Twenty-seven patients were included. Patients were pretreated with a median of two lines of cisplatin-based chemotherapy (range 1-5). Sixteen patients were absolute refractory. Five patients had relapsed after high-dose chemotherapy plus stem-cell support. There were no complete responses but there was one marker positive partial response and nine disease stabilization (34, 6%). After a median follow-up of 65 months, two patients are disease-free survivors. Main toxicity was leucocytopenia grade 3/4 in 30% of the patients. CONCLUSION: Combination chemotherapy with oxaliplatin and paclitaxel is feasible with acceptable toxicity and may be effective if combined with additional treatment in patients with CDDP refractory GCT. PMID- 18385204 TI - Age-specific trends in mammographic density: the Minnesota Breast Cancer Family Study. AB - Mammographic density is a strong risk factor for breast cancer, yet few studies have evaluated density trends, and associated factors, over time. The authors retrieved and digitized mammograms (> or =1 per woman) imaged in 1990-2003 to evaluate percent density (PD) in the Minnesota Breast Cancer Family cohort. Multivariable-adjusted, mixed-effects, repeated-measures models incorporating a natural cubic spline provided estimates of nonlinear trends in PD with age and were used to examine association with covariates. Overall, 5,698 mammograms from 1,689 women with covariate information were digitized. In descriptive analyses, the highest median PD was 33.1% (interquartile range, 21.8%; n = 230) among premenopausal women, 31.0% (interquartile range, 23.2%; n = 175) among women who transitioned from pre- to postmenopause, and 18.7% (interquartile range, 22.2%; n = 1,284) among postmenopausal women. On average, premenopausal compared with postmenopausal women had 1.9% (p = 0.001) higher PD. In repeated-measures analyses, greater declines in PD occurred with menopause and among women with higher baseline PD; current postmenopausal hormone use and higher body mass index modified these declines (p interaction < 0.001). No significant modification of the density change with age was seen with parity/age at first birth, age at menarche, oral contraceptive use, family history of breast or ovarian cancer in a first- or second-degree relative, educational level, smoking status, or alcohol intake were observed. These data suggest that menopause, baseline PD, postmenopausal hormone use, and body mass index predict changes in mammographic density trends during adult life. PMID- 18385205 TI - Invited commentary: assessing breast density change--lessons for future studies. AB - Breast density is one of the strongest predictors of breast cancer risk, and quantitative measurement is fairly reproducible. However, to study change in breast density, other issues should also be considered. Most studies of breast density have relied on one assessment, yet the mammographic features of the breast that constitute breast density change with age and/or menopause. When measuring breast density change, issues related to assessment are of greater concern. In addition, because age-period and cohort effects are codefined, evaluation of age trends must also consider the possible explanations of period and cohort effects. The prevalence of different factors affecting breast density changed dramatically over the last 50 years. In this issue of the Journal (Am J Epidemiol 2008;167:1027-1036), Kelemen et al. evaluate how factors known to be related to breast density influence breast density change with age. These authors are to be complimented on their detailed analysis and consideration of many of these issues. They not only describe the averaged effects of age on breast density changes but also consider whether patterns of density change differ for women with different exposure histories. PMID- 18385206 TI - Glomerular filtration rate, albuminuria, and risk of cardiovascular and all-cause mortality in the US population. AB - Decreased glomerular filtration rate (GFR) and albuminuria are used in combination to define chronic kidney disease, but their separate and combined effects on cardiovascular and all-cause mortality have not been studied in the general population. The linked mortality file of the Third National Health and Nutrition Examination Survey includes data from 13 years of follow-up (1988-2000) for 14,586 US adults. The authors estimated GFR from standardized serum creatinine levels. Albuminuria was defined by the urinary albumin:creatinine ratio. Incidence rate ratios (IRRs) were adjusted for major cardiovascular disease risk factors and C-reactive protein. Lower estimated GFR was associated with higher risks of cardiovascular and all-cause mortality overall and within every albuminuria category. Likewise, increasing albuminuria was associated with higher risk of estimated GFR overall and within every category. When estimated GFR and albuminuria were examined simultaneously, a 10-ml/minute/1.73 m(2) lower estimated GFR (among persons with estimated GFR <60 ml/minute/1.73 m(2)) was associated with an IRR of 1.29 (95% confidence interval: 1.06, 1.55) for cardiovascular mortality and a doubling of albuminuria was associated with an IRR of 1.06 (95% confidence interval: 1.04, 1.08) for cardiovascular mortality. The authors conclude that moderately decreased estimated GFR and albuminuria independently predict cardiovascular and all-cause mortality in the general population. These data support recent recommendations defining chronic kidney disease and stratifying subsequent risks based on both decreased GFR and albuminuria. PMID- 18385207 TI - Principles of antipsychotic prescribing for policy makers, circa 2008. Translating knowledge to promote individualized treatment. AB - Findings from 2 pivotal government-funded studies of comparative antipsychotic effectiveness undermine assumptions about the marked superiority of the more expensive second-generation "atypical" medications in comparison to the less expensive first-generation "typical" drugs. Because this assumption was the basis for the almost universal recommendation that these newer antipsychotics be used preferentially resulting in a 10-fold increase in state governmental expenditures on this class of medications over the past decade, a reassessment of policy is called for. To address the issue, the Medical Directors Council of the National Association of State Mental Health Program Directors critically reviewed findings of these studies in the context of other data and considered policy implications in the light of the obligations of state government to make available best possible and individually optimized treatment that is cost-effective. The Medical Directors Council unanimously adopted a set of recommendations to promote appropriate access, efficient utilization, and best practice use. We present our policy statement, in which we provide a succinct background, articulate general principles, and describe a set of 4 broad recommendations. We then summarize our understanding of the current state of knowledge about comparative antipsychotic effectiveness, best antipsychotic practice, and considerations for state policy that represent the basis of our position statement. PMID- 18385209 TI - Physiologically based pharmacokinetic modeling of chloroethane disposition in mice, rats, and women. AB - Chloroethane was observed in a chronic cancer bioassay to be a mouse-specific uterine carcinogen at a single high inhaled concentration (15,000 ppm). Although high incidence occurred in the female mouse (86%), no uterine tumor increases were observed in female rats. Chloroethane is a weak alkylating agent and has low acute toxicity. No genotoxicity potential has been observed below 40,000 ppm. Chloroethane is eliminated from the body by pulmonary exhalation and metabolically by oxidation via cytochrome P-450 (likely producing acetaldehyde) and conjugation with glutathione (GSH). The mode of action for the mouse-specific uterine tumors is not definitively known and could involve parent chemical and/or metabolite(s). A physiologically based pharmacokinetic (PBPK) model for chloroethane disposition in the rat was developed previously, but no such models have been described for mice or humans. For the work reported here, the existing PBPK model for chloroethane in rats was expanded and refined, and PBPK models for chloroethane disposition in mice and humans were developed to allow species comparisons of internal dosimetry and for possible insights into the carcinogenic mode of action. The amounts metabolized via glutathione-S-transferase (GST) versus cytochrome P-450, and the total amount of chloroethane absorbed, were most consistent with the observations made concerning uterine tumors, with amounts metabolized via GST providing the larger quantitative difference between the two rodent species. Choice of the most relevant dose metric for risk assessments involving uterine tumors in mice will require pharmacodynamic considerations in the mode of action in addition to the pharmacokinetic differences reported here. PMID- 18385202 TI - Extracapsular tumor spread and the risk of local, axillary and supraclavicular recurrence in node-positive, premenopausal patients with breast cancer. AB - BACKGROUND: Extracapsular tumor spread (ECS) has been identified as a possible risk factor for breast cancer recurrence, but controversy exists regarding its role in decision making for regional radiotherapy. This study evaluates ECS as a predictor of local, axillary, and supraclavicular recurrence. PATIENTS AND METHODS: International Breast Cancer Study Group Trial VI accrued 1475 eligible pre- and perimenopausal women with node-positive breast cancer who were randomly assigned to receive three to nine courses of classical combination chemotherapy with cyclophosphamide, methotrexate, and fluorouracil. ECS status was determined retrospectively in 933 patients based on review of pathology reports. Cumulative incidence and hazard ratios (HRs) were estimated using methods for competing risks analysis. Adjustment factors included treatment group and baseline patient and tumor characteristics. The median follow-up was 14 years. RESULTS: In univariable analysis, ECS was significantly associated with supraclavicular recurrence (HR = 1.96; 95% confidence interval 1.23-3.13; P = 0.005). HRs for local and axillary recurrence were 1.38 (P = 0.06) and 1.81 (P = 0.11), respectively. Following adjustment for number of lymph node metastases and other baseline prognostic factors, ECS was not significantly associated with any of the three recurrence types studied. CONCLUSIONS: Our results indicate that the decision for additional regional radiotherapy should not be based solely on the presence of ECS. PMID- 18385208 TI - An aryl hydrocarbon receptor repressor from Xenopus laevis: function, expression, and role in dioxin responsiveness during frog development. AB - Xenopus laevis and other frogs are extremely insensitive to the toxicity of xenobiotic ligands of the aryl hydrocarbon receptor (AHR), including 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). Premetamorphic life stages are especially insensitive, and they are reported to be refractory to induction of Cytochrome P4501As, which are readily induced in older animals. The AHR repressor (AHRR) is a member of the AHR gene family. AHRR expression is induced by TCDD; it then represses AHR in an apparent negative feedback loop. In this study, we sought to test the hypothesis that constitutive AHRR expression underlies the lack of TCDD responsiveness in frog early life stages. We determined the sequence of an AHRR complimentary DNA encoding an 85.3-kDa protein sharing 52-55% identity with the bHLH/PAS domains of other AHRRs. In transient transfection assays, X. laevis AHRR inhibited TCDD-induced reporter gene expression mediated by either X. laevis AHR paralog, AHR1alpha or AHR1beta. AHRR messenger RNA was expressed at low levels in embryos (Nieuwkoop-Faber stage 33-38; approximately 52 h.p.f.) and was induced approximately twofold following TCDD exposure (42 ng/g wet weight). In contrast, AHRR exhibited higher constitutive expression and was induced more than threefold in tadpoles at stage 52-55 (prometamorphic; approximately 4 weeks postfertilization) and in isolated viscera of stage 62 tadpoles (in the metamorphic climax; approximately 7 weeks postfertilization). Although the magnitude of induction was smaller, the temporal pattern of AHRR expression and inducibility resembled that of CYP1A6. Thus, attenuated transcriptional activation of AHR target genes and low TCDD toxicity in X. laevis embryos cannot be explained by constitutive, high-level expression of AHRR. PMID- 18385211 TI - Comments on "Evaluation of estrogenic activities of aquatic herbicides and surfactants using a rainbow trout vitellogenin assay". PMID- 18385212 TI - Bilateral popliteal artery occlusion in a competitive bike rider: case report and clinical review. AB - Since its first description in 1879, popliteal artery entrapment syndrome remains a debilitating condition, which frequently affects young active people. Increased awareness of popliteal artery entrapment syndrome combined with improvements in investigative modalities has resulted in a more frequent diagnosis of this eminently treatable condition. In this article, a rare case of bilateral popliteal artery entrapment syndrome in a physically active 33-year-old man precipitated by competitive Bicycle Moto-Cross riding is presented. A higher index of suspicion for popliteal artery entrapment syndrome is recommended as the underlying pathology in a young active person with symptoms of lower limb claudication. Popliteal artery entrapment syndrome should be investigated with thorough radiological assessment prior to any therapeutic intervention, which is often fraught with difficulty due to chronically diseased vasculature and inherent anatomical anomalies. PMID- 18385213 TI - B56beta, a regulatory subunit of protein phosphatase 2A, interacts with CALEB/NGC and inhibits CALEB/NGC-mediated dendritic branching. AB - The development of dendritic arbors is critical in neuronal circuit formation, as dendrites are the primary sites of synaptic input. Morphologically specialized dendritic protrusions called spines represent the main postsynaptic compartment for excitatory neurotransmission. Recently, we demonstrated that chicken acidic leucine-rich epidermal growth factor (EGF) -like domain-containing brain protein/neuroglycan C (CALEB/NGC), a neural member of the EGF family, mediates dendritic tree and spine complexity but that the signaling pathways in the respective processes differ. For a more detailed characterization of these signal transduction pathways, we performed a yeast two-hybrid screen to identify proteins that interact with CALEB/NGC. Our results show that B56beta, a regulatory subunit of protein phosphatase 2A, interacts with CALEB/NGC and inhibits CALEB/NGC-mediated dendritic branching but not spine formation. Binding of B56beta to CALEB/NGC was confirmed by several biochemical and immunocytochemical assays. Using affinity chromatography and mass spectrometry, we demonstrate that the whole protein phosphatase 2A trimer, including structural and catalytic subunits, binds to CALEB/NGC via B56beta. We show that CALEB/NGC induces the phosphorylation of Akt in dendrites. Previously described to interfere with Akt signaling, B56beta inhibits Akt phosphorylation and Akt dependent dendritic branching but not Akt-independent spine formation induced by CALEB/NGC. Our results contribute to a better understanding of signaling specificity leading to neuronal process differentiation in sequential developmental events. PMID- 18385214 TI - Neoarteries grown in vivo using a tissue-engineered hyaluronan-based scaffold. AB - Vascular tissue engineering has emerged as a promising technology for the design of an ideal, responsive, living conduit with properties similar to that of native tissue. The missing link in tissue-engineered blood vessels is elastin biosynthesis. Several biomaterials are currently used but few support elastin biosynthesis in a 3-D array. In previous studies, we demonstrated that a hyaluronan-based scaffold (HYAFF-11) grafted in the infrarenal rat aorta successfully guided the complete regeneration of a well-functioning small diameter (2 mm) neoartery. The aim of the present study was to test the ability of HYAFF-11 biodegradable grafts to develop into neovessels of larger size (4 mm) in a porcine model, focusing on extracellular matrix (ECM) remodeling and elastin biosynthesis. HYAFF-11 tubes (diameter 4 mm, length 5 cm) were implanted in an end-to-end fashion in the common carotid artery. Grafts were analyzed for patency with a Duplex scan every 15 days. ECM components were evaluated by histological and molecular biological methods. All the animals survived the observation period without complications. Intimal hyperplasia (initiating at the anastomotic site) and graft thrombosis led to 3 cases of partial or complete occlusion, as demonstrated by histological examination. There were no signs of stenoses or aneurysms in the remaining grafts. After 5 months, the biomaterial was almost completely degraded and replaced by a neoartery segment composed of mature smooth muscle cells, collagen, and elastin fibers organized in layers and was completely covered on the luminal surface by endothelial cells (vWF(+)). Whereas in previous small animal studies, patency rates were not optimal, those obtained in the present study using hyaluronan-based grafts of larger size confirmed the ability of these constructs to guide the development of a well-functioning neoartery, with the remarkable additional attribute of facilitating the formation of organized layers of elastin fibers. PMID- 18385215 TI - Analogous oligo-acyl-lysines with distinct antibacterial mechanisms. AB - Bactericidal properties were recently shown to emerge from hydrophobicity and charge buildup in oligo-acyl-lysine (OAK) peptide mimetics. Toward understanding the attributes that govern the activity of this novel antimicrobial system, we compared the functional and mechanistic properties of a known octamer and a newly generated hexamer analog. The data provide strong evidence for multiple similarities that included high tissue stability, low hemolysis, large-spectrum antibacterial activity in vitro, and the ability to prevent Escherichia coli induced mortality in vivo. Despite these similarities, however, the octamer mode of action involved membrane disruption, unlike the hexamer, which acted predominantly through inhibition of DNA functions with characteristically slower bactericidal kinetics. Collectively, the data support the view that the analogous OAKs induced bacterial death by distinct mechanisms and further suggest that relatively minor differences in the sequence of host defense peptides are responsible for selecting one mechanism over another, possibly in conjunction with differential binding affinities to the external and/or cytoplasmic membrane. PMID- 18385216 TI - Aortic angiosarcoma mimicking large-vessel vasculitis: a diagnostic dilemma. PMID- 18385217 TI - Management of accelerated pulmonary nodulosis following etanercept therapy in a patient with rheumatoid arthritis. PMID- 18385218 TI - Plastid-derived genes in the nonphotosynthetic alveolate Oxyrrhis marina. AB - Reconstructing the history of plastid acquisition and loss in the alveolate protists is a difficult problem because our knowledge of the distribution of plastids in extant lineages is incomplete due to the possible presence of cryptic, nonphotosynthetic plastids in several lineages. The discovery of the apicoplast in apicomplexan parasites has drawn attention to this problem and, more specifically, to the question of whether many other nonphotosynthetic lineages also contain cryptic plastids or are derived from plastid-containing ancestors. Oxyrrhis marina is one such organism: It is a heterotrophic, early branching member of the dinoflagellate lineage for which there is no evidence of a plastid. To investigate the possibility that O. marina is derived from a photosynthetic ancestor, we have generated and analyzed a large-scale EST database and searched for evidence of plastid-derived genes. Here, we describe 8 genes whose phylogeny shows them to be derived from plastid-targeted homologues. These genes encode proteins from several pathways known to be localized in the plastids of other algae, including synthesis of tetrapyrroles, isoprenoids, and amino acids, as well as carbon metabolism and oxygen detoxification. The 5' end of 5 cDNAs were also characterized using cap-dependent or spliced leader-mediated reverse transcriptase-polymerase chain reaction, revealing that at least 4 of these genes have retained leaders that are similar in nature to the plastid targeting signals of other secondary plastids, suggesting that these proteins may be targeted to a cryptic organelle. At least 2 genes do not encode such leaders, and their products may presently function in the cytosol. Altogether, the presence of plastid-derived genes in O. marina shows that its ancestors contained a plastid, and the pathways represented by the genes and presence of targeting signals on at least some of the genes further suggests that a relict organelle may still exist to fulfill plastid metabolic functions. PMID- 18385219 TI - Parallel rate heterogeneity in chloroplast and mitochondrial genomes of Brazil nut trees (Lecythidaceae) is consistent with lineage effects. AB - We investigated whether relative rates of divergence were correlated between the mitochondrial and chloroplast genomes as expected under lineage effects or were genome specific as expected with locus-specific effects. Five mitochondrial noncoding regions (nad1B_C, nad4exon1_2, nad7exon2_3, nad7exon3_4, and rps14-cob) for 21 samples from Lecythidaceae were sequenced. Three chloroplast regions (rpl20-5'rps12, trnS-trnG, and psbA-trnH) were sequenced to expand the taxa in an existing data set. Absolute rates of nucleotide and insertion and deletion (indel) changes were 13 times faster in the chloroplast genome than in the mitochondrial genome. Similar indel length frequency distributions for both organelles suggested that common mechanisms were responsible for generating indels. Molecular clock tests applied to phylogenetic trees estimated from mitochondrial and chloroplast sequences revealed global rate heterogeneity of nucleotide substitution. Maximum likelihood and Tajima's 1D relative rate tests show that Lecythis zabucajo exhibited a rate acceleration for both the mitochondrial and chloroplast sequences. Whereas Eschweilera romeu-cardosoi showed a significant rate slowdown for chloroplast sequences, the mitochondrial sequences for 3 Eschweilera taxa showed evidence for a rate slowdown only when compared with L. zabucajo. Significant rate heterogeneity was also observed for indel changes in the mitochondrial genome but not for the chloroplast. The lack of mitochondrial nucleotide changes for some taxa as well as chloroplast indel homoplasy may have limited the power of relative rate tests to detect rate variation. Relative ratio tests consistently indicated rate proportionality among branch lengths between the mitochondrial and chloroplast phylogenetic trees. The relative ratio tests showed that taxa possessing rate heterogeneity had parallel relative divergence rates in both mitochondrial and chloroplast sequences as expected under lineage effects. A neutral replication-dependent model of rate heterogeneity for both nucleotide and indel changes provides a simple explanation for common patterns of rate heterogeneity across the 2 organelle genomes in Lecythidaceae. The lineage effects observed here were uncoupled from annual/perennial habit because all the species from this study are perennial. PMID- 18385220 TI - Local adaptation and vector-mediated population structure in Plasmodium vivax malaria. AB - Plasmodium vivax in southern Mexico exhibits different infectivities to 2 local mosquito vectors, Anopheles pseudopunctipennis and Anopheles albimanus. Previous work has tied these differences in mosquito infectivity to variation in the central repeat motif of the malaria parasite's circumsporozoite (csp) gene, but subsequent studies have questioned this view. Here we present evidence that P. vivax in southern Mexico comprised 3 genetic populations whose distributions largely mirror those of the 2 mosquito vectors. Additionally, laboratory colony feeding experiments indicate that parasite populations are most compatible with sympatric mosquito species. Our results suggest that reciprocal selection between malaria parasites and mosquito vectors has led to local adaptation of the parasite. Adaptation to local vectors may play an important role in generating population structure in Plasmodium. A better understanding of coevolutionary dynamics between sympatric mosquitoes and parasites will facilitate the identification of molecular mechanisms relevant to disease transmission in nature and provide crucial information for malaria control. PMID- 18385221 TI - Unicellular Ca2+ signaling 'toolkit' at the origin of metazoa. AB - Ca(2+) signaling pathways control many physiological processes in almost all types of animal cells such as fertilization, muscle contraction, hormone release, and learning and memory. Each animal cell type expresses a unique group of molecules from the Ca(2+) signaling 'toolkit' to control spatiotemporal patterns of Ca(2+) signaling. It is generally believed that the complex Ca(2+) signaling 'toolkit' has arisen from the ancestral multicellular organisms to fit unique physiological roles of specialized cell types. Here, we demonstrate for the first time the presence of an extensive Ca(2+) signaling 'toolkit' in the unicellular choanoflagellate Monosiga brevicollis. Choanoflagellates possess homologues of various types of animal plasma membrane Ca(2+) channels including the store operated channel, ligand-operated channels, voltage-operated channels, second messenger-operated channels, and 5 out of 6 animal transient receptor potential channel families. Choanoflagellates also contain homologues of inositol 1,4,5 trisphosphate receptors. Furthermore, choanoflagellates master a complete set of Ca(2+) removal systems including plasma membrane and sarco/endoplasmic reticulum Ca(2+) ATPases and homologues of 3 animal cation/Ca(2+) exchanger families. Therefore, a complex Ca(2+) signaling 'toolkit' might have evolved before the emergence of multicellular animals. PMID- 18385222 TI - Novel transcriptome patterns accompany evolutionary restoration of defective social development in the bacterium Myxococcus xanthus. AB - Evolutionary trait losses can be restored by direct reversion or by compensatory pathways. Upon starvation, the bacterium Myxococcus xanthus develops into spore bearing fruiting bodies, but this ability can be rapidly lost during evolution. Some developmentally defective strains of M. xanthus "cheat" on proficient strains during development by superior sporulation in mixed cultures. Here, we examine transcriptomic patterns accompanying the evolution of a cheater (obligate cheater [OC]) to a developmentally competent strain (PX) by a single mutation. Using quantitative real-time-polymerase chain reaction analysis of 5 genes essential for development, we initially show that restoration of development in strain PX was associated with increased expression of 4 of these genes, not only relative to OC but also relative to the developmentally proficient ancestor of both OC and PX (wild type [WT]). Global transcriptome analyses showed further that developmental expression of well more than 100 genes differ significantly between PX and the proficient WT ancestor. Moreover, the expression profile of PX was found to differ from that of WT more than does that of the defective intermediate strain OC. These results show that the restoration of a complex trait is accompanied by novel expression patterns across a large number and wide variety of genes, rather than by a large-scale return to ancestral expression patterns. PMID- 18385223 TI - Prevalence and risk factors for carcinogenic human papillomavirus infections in rural Rakai, Uganda. AB - OBJECTIVE: To investigate self-administered vaginal swabs for assessing prevalence and correlates of carcinogenic human papillomavirus (HPV) infection in rural Rakai, Uganda. METHODS: 1003 sexually experienced women enrolled in a community cohort provided self-administered vaginal swabs collected at annual, home-based surveys. Carcinogenic HPV prevalence, adjusted odds ratios (AOR), 95% confidence intervals (CI) and associated risk factors were determined. RESULTS: Carcinogenic HPV prevalence was 19.2%: 46.6% among HIV positive and 14.8% among HIV negative women (p<0.001). Type-specific prevalence ranged from 2.0% (HPV 16 and 52) to 0.2% (HPV 31). Age-specific HPV prevalence decreased significantly (p<0.001) among HIV negative women; however, the decrease among HIV positive women was not as pronounced (p = 0.1). Factors independently associated with carcinogenic HPV infection were HIV (AOR 4.82, CI 3.10 to 7.53), age (AOR 4.97, 95% CI 2.19 to 11.26 for 15-19 year olds compared to 40+ years), more than two sex partners in the past year (AOR 2.21, CI 1.10 to 4.43) and self-reported herpes zoster, candidiasis or tuberculosis (AOR 4.52, CI 1.01 to 20.31). Married women were less likely to have prevalent carcinogenic HPV (AOR 0.46, CI 0.30 to 0.70). CONCLUSIONS: HPV prevalence and correlates measured using self administered vaginal swabs were similar to studies that use cervical samples. Thus, self-collection can be used as a substitute for cervical specimens and provide an important tool for research in populations unwilling to undergo pelvic exam. PMID- 18385224 TI - Self-report is a valid measure of circumcision status in homosexual men. AB - OBJECTIVE: Misreporting of circumcision status may affect observed relationships between circumcision status and HIV or other sexually transmissible infections. As no data exist on the validity of self-reported circumcision status among homosexual men, we investigated the agreement between self-report and examination findings in a subgroup of participants in the Health in Men (HIM) study in Sydney, Australia. METHODS: A subgroup of 240 participants in the community based HIM cohort study attending annual interview agreed to a brief genital examination by a trained study nurse who was unaware of their previous self-reported circumcision status. RESULTS: Five participants reported being uncircumcised at baseline but were classified as circumcised on examination. All participants who self-reported being circumcised were found on examination to be circumcised. Three cases in which the examining study nurse was unsure of participants' circumcision status were excluded. Of the remaining 237 participants, 155 (65.4%) were classified as circumcised on examination, including five men who self identified as uncircumcised. Compared with examination, self-reported circumcision status resulted in a sensitivity of 96.8%, specificity of 100%, positive predictive value of 100% and negative predictive value of 94.3%. The overall agreement between circumcision status on examination and self-report was 97.9% (kappa score, 0.95; p<0.001) CONCLUSION: Self-report was a valid measure of circumcision status in this group of predominantly Anglo gay-community-attached men. We believe our findings can be generalised to similarly aged gay-community attached men in other developed countries. PMID- 18385225 TI - The burden of asymptomatic sexually transmitted infections among men in Carletonville, South Africa: implications for syndromic management. AB - OBJECTIVES: To determine the prevalence of sexually transmitted infections (STI) and HIV among men attending an outreach STI/HIV screening service in informal settlements in South Africa over a 3-month period. METHODS: A mobile clinic van was utilised to offer men: (1) urine screening for gonorrhoea, trichomoniasis, chlamydial and Mycoplasma genitalium infections; (2) serological screening for syphilis and herpes simplex virus type 2 (HSV-2); (3) on-site HIV voluntary counselling and testing. Urethritis pathogens were detected by molecular methods. HIV serostatus was determined using rapid tests. Demographic, sexual behaviour and clinical data were recorded on a nurse-administered questionnaire. Statistical analysis utilised the chi(2) test. RESULTS: 309 men attended the service; 304 (98%) requested serological screening for syphilis and HSV-2, 301 (97%) underwent urine-based screening for urethritis pathogens and 269 (87%) had an HIV test. Over 90% of men were asymptomatic for STI. Gonorrhoea was more prevalent in the symptomatic group (p<0.001); there were no significant differences in the prevalence of other urethritis pathogens between the groups. The total number of infections with each urethritis pathogen was highest in the asymptomatic group with twice as much gonorrhoea, 25 times as much chlamydial infection, six times as much trichomoniasis and nine times as much M genitalium infection compared with the symptomatic group. The overall HIV prevalence among clinic attendees was 29.7%. CONCLUSIONS: The uptake of both STI and HIV testing was high among men attending the service. The relatively high burden of both STI and HIV among the male clinic attendees has implications for the transmission of HIV. PMID- 18385226 TI - Modelling the cost per ulcer treated of incorporating episodic treatment for HSV 2 into the syndromic algorithm for genital ulcer disease. AB - BACKGROUND: The proportion of genital ulcer disease (GUD) due to herpes simplex virus type-2 (HSV-2) has increased in sub-Saharan Africa. The most recent 2003 WHO syndromic GUD algorithm includes antiviral treatment for HSV-2 for anyone with "typical" symptoms/signs, and suggests that all GUD patients receive treatment for HSV-2 in settings where HSV-2 GUD aetiology is greater than 30%. The previous algorithm (1994) only targeted Haemophilus ducreyi (HD) and Treponema pallidum (TP). METHODS: A static deterministic model was used to compare the cost per ulcer treated of using the 1994 and 2003 algorithms amongst individuals presenting with GUD, with sensitivity analyses for different economic and epidemiological scenarios. RESULTS: Except when the proportion of ulcers due to HD/TP (defined as ulcer prevalence) is high (>40%), and HSV-2 ulcer prevalence is low (<30%), the 2003 algorithm should result in more patients receiving the correct treatment (correct drugs for the syndrome) than the 1994 algorithm, and it will cost less per ulcer treated if HSV-2 treatment costs less than US$2. Greatest impact in terms of ulcers treated is achieved with the 2003 algorithm if HSV-2 treatment is given to all GUD patients. The incremental and/or relative cost per ulcer treated of doing this, compared to only treating those with typical symptoms/signs, is reduced if the HSV-2 ulcer prevalence is high and/or the HSV-2 treatment cost or sensitivity of HSV-2 ulcer diagnosis (using symptoms/signs) is low. CONCLUSIONS: In certain scenarios, including HSV-2 treatment can increase the number of ulcers treated and reduce the cost per ulcer treated of GUD syndromic management. PMID- 18385227 TI - Plasmacytoid dendritic cell dynamics and alpha interferon production during Simian immunodeficiency virus infection with a nonpathogenic outcome. AB - We addressed the role of plasmacytoid dendritic cells (PDC) in protection against AIDS in nonpathogenic simian immunodeficiency virus (SIVagm) infection in African green monkeys (AGMs). PDC were monitored in blood and lymph nodes (LNs) starting from day 1 postinfection. We observed significant declines in blood during acute infection. However, PDC then returned to normal levels, and chronically infected AGMs showed no decrease of PDC in blood. There was a significant increase of PDC in LNs during acute infection. Blood PDC displayed only weak alpha interferon (IFN-alpha) responses to TLR9 agonist stimulation before infection. However, during acute infection, both blood and LN PDC showed a transiently increased propensity for IFN-alpha production. Bioactive IFN-alpha was detected in plasma concomitant with the peak of viremia, though levels were only low to moderate in some animals. Plasma interleukin 6 (IL-6) and IL-12 were not increased. In conclusion, PDC were recruited to the LNs and displayed increased IFN-alpha production during acute infection. However, increases in IFN-alpha were transient. Together with the lack of inflammatory cytokine responses, these events might play an important role in the low level of T-cell activation which is associated with protection against AIDS in nonpathogenic SIVagm infection. PMID- 18385228 TI - Structural and functional constraints limit options for cytotoxic T-lymphocyte escape in the immunodominant HLA-B27-restricted epitope in human immunodeficiency virus type 1 capsid. AB - Control of human immunodeficiency virus type 1 (HIV-1) by HLA-B27-positive subjects has been linked to an immunodominant CD8(+) cytotoxic T-lymphocyte (CTL) response targeting the conserved KK10 epitope (KRWIILGLNK(263-272)) in p24/Gag. Viral escape in KK10 typically occurs through development of an R(264)K substitution in conjunction with the upstream compensatory mutation S(173)A, and the difficulty of the virus to escape from the immune response against the KK10 epitope until late in infection has been associated with slower clinical progression. Rare alternative escape mutations at R(264) have been observed, but factors dictating the preferential selection of R(264)K remain unclear. Here we illustrate that while all observed R(264) mutations (K, G, Q, and T) reduced peptide binding to HLA-B27 and impaired viral replication, the replicative defects of the alternative mutants were actually less pronounced than those for R(264)K. Importantly, however, none of these mutants replicated as well as an R(264)K variant containing the compensatory mutation S(173)A. In assessing the combined effects of viral replication and CTL escape using an in vitro coculture assay, we further observed that the compensated R(264)K mutant also displayed the highest replication capacity in the presence of KK10-specific CTLs. Comparisons of codon usage for the respective variants indicated that generation of the R(264)K mutation may also be favored due to a G-to-A bias in nucleotide substitutions during HIV-1 replication. Together, these data suggest that the preference for R(264)K is due primarily to the ability of the S(173)A-compensated virus to replicate better than alternative variants in the presence of CTLs, suggesting that viral fitness is a key contributor for the selection of immune escape variants. PMID- 18385229 TI - Paucity of CD4+ CCR5+ T cells may prevent transmission of simian immunodeficiency virus in natural nonhuman primate hosts by breast-feeding. AB - Simian immunodeficiency virus (SIV) persistence in wild populations of African nonhuman primates (NHPs) may occur through horizontal and vertical transmission. However, the mechanism(s) and timing of the latter type of transmission have not been investigated to date. Here we present the first study of SIV transmissibility by breast-feeding in an African NHP host. Six mandrill dames were infected with plasma containing 300 50% tissue culture infective doses of SIVmnd-1 on the day after delivery. All female mandrills became infected, as demonstrated by both plasma viral loads (VLs) and anti-SIVmnd-1 seroconversion. Neither fever nor lymphadenopathy was observed. At the peak of SIVmnd-1 viral replication (days 7 to 10 postinoculation), plasma VLs were high (8 x 10(6) to 8 x 10(8) RNA copies/ml) and paralleled the high VLs in milk (4.7 x 10(4) to 5.6 x 10(5) RNA/ml). However, at the end of the breast-feeding period, after 6 months of follow-up, no sign of infection was observed for the offspring. Later on, during a 4-year follow-up examination, two of the offspring showed virological evidence of SIVmnd-1 infection. Both animals seroconverted at least 6 months after the interruption of lactation. In conclusion, despite extensive viral replication in mandrill mothers and high levels of free virus in milk, no SIVmnd 1 transmission was detectable at the time of breast-feeding or during the following months. Since we observed a markedly lower expression of CCR5 on the CD4(+) T cells of young mandrills and African green monkeys than on those of adults, we propose that low levels of this viral coreceptor on CD4(+) T cells may be involved in the lack of breast-feeding transmission in natural hosts of SIVs. PMID- 18385230 TI - Cyclophilin A is an essential cofactor for hepatitis C virus infection and the principal mediator of cyclosporine resistance in vitro. AB - Cyclosporine (CsA) and its derivatives potently suppress hepatitis C virus (HCV) replication. Recently, CsA-resistant HCV replicons have been identified in vitro. We examined the dependence of the wild-type and CsA-resistant replicons on various cyclophilins for replication. A strong correlation between CsA resistance and reduced dependency on cyclophilin A (CyPA) for replication was identified. Silencing of CyPB or CyPC expression had no significant effect on replication, whereas various forms of small interfering RNA (siRNA) directed at CyPA inhibited HCV replication of wild-type but not CsA-resistant replicons. The efficiency of a particular siRNA in suppressing CyPA expression was correlated with its potency in inhibiting HCV replication, and expression of an siRNA-resistant CyPA cDNA rescued replication. In addition, an anti-CyPA antibody blocked replication of the wild-type but not the resistant replicon in an in vitro replication assay. Depletion of CyPA alone in the CsA-resistant replicon cells eliminated CsA resistance, indicating that CyPA is the chief mediator of the observed CsA resistance. The dependency on CyPA for replication was observed for both genotype (GT) 1a and 1b replicons as well as a GT 2a infectious virus. An interaction between CyPA and HCV RNA as well as the viral polymerase that is sensitive to CsA treatment in wild-type but not in resistant replicons was detected. These findings reveal the molecular mechanism of CsA resistance and identify CyPA as a critical cellular cofactor for HCV replication and infection. PMID- 18385231 TI - Characterization of a rhesus monkey calicivirus representing a new genus of Caliciviridae. AB - In this study, we report the characterization of a novel calicivirus (CV), the Tulane virus (TV), which was isolated from stool samples of captive juvenile rhesus macaques (Macaca mulatta) of the Tulane National Primate Research Center. The complete genome of TV contains 6,714 nucleotides plus a poly(A) tail and is organized into three open reading frames (ORFs) that encode the nonstructural (NS) polyprotein (ORF1); the capsid protein (ORF2), with an estimated molecular mass of 57.9 kDa; and a possible minor structural protein (ORF3), with an isoelectric point (pI) of 10.0 and a calculated molecular mass of 22.8 kDa. The NS polyprotein revealed all typical CV amino acid motifs, including GXXGXGKT (NTPase), EYXEX (Vpg), GDCG (protease), and GLPSG and YGDD (polymerase). Phylogenetic trees constructed for the NS polyprotein, NTPase, protease, polymerase, and capsid protein sequences consistently placed the TV on a branch rooted with Norovirus, but with distances equal to those between other genera. The TV can be cultured in a monkey kidney cell line (LLC-MK2) with the appearance of typical cytopathic effect. TV exhibits a typical CV morphology, with a diameter of 36 nm, and has a buoyant density of 1.37 g/ml. According to these physicochemical and genetic characteristics, TV represents a new CV genus for which we propose the name "Recovirus" (rhesus enteric CV). Although the pathogenicity of TV in rhesus macaques remains to be elucidated, the likelihood of TV causing intestinal infection and the availability of a tissue culture system make this virus a valuable surrogate for human CVs. PMID- 18385232 TI - Roles of LEF-4 and PTP/BVP RNA triphosphatases in processing of baculovirus late mRNAs. AB - The baculovirus Autographa californica nucleopolyhedrovirus encodes two proteins with RNA triphosphatase activity. Late expression factor LEF-4, which is an essential gene, is a component of the RNA polymerase and also encodes the RNA capping enzyme guanylyltransferase. PTP/BVP is also an RNA triphosphatase, but is not essential for viral replication, possibly because its activity is redundant to that of LEF-4. To elucidate the role of these proteins in mRNA cap formation, a mutant virus that lacked both RNA triphosphatase activities was constructed. Infection studies revealed that the double-mutant virus was viable and normal with respect to the production of budded virus. Pulse-labeling studies and immunoblot analyses showed that late gene expression in the double mutant was equivalent to that in the wild type, while polyhedrin expression was slightly reduced. Direct analysis of the mRNA cap structure indicated no alteration of cap processing in the double mutant. Together, these results reveal that baculoviruses replicate and express their late genes at normal levels in the absence of its two different types of RNA triphosphatases. PMID- 18385233 TI - West Nile virus entry requires cholesterol-rich membrane microdomains and is independent of alphavbeta3 integrin. AB - West Nile virus (WNV) has been the leading cause of viral encephalitis in the United States since 1999. The endocytic processes involved in the internalization of infectious WNV by various cell types are not well characterized, and the involvement of cholesterol-rich membrane microdomains, or lipid rafts, in the life cycle of WNV has not been investigated previously. In this study, we found that the depletion of cellular cholesterol levels by brief treatment with methyl beta-cyclodextrin resulted in a 100-fold reduction of the titers of infectious WNV released into the culture supernatant, as well as a reduction in the number of WNV genome copies in the cholesterol-depleted cells. The addition of exogenous cholesterol to cholesterol-depleted cells reversed this effect. Cholesterol depletion postinfection did not affect WNV growth, suggesting that the effect occurs at the level of WNV entry. We also showed that while WNV entry did not require alphavbeta3 integrin and focal adhesion kinase, WNV particles failed to be internalized by cholesterol-depleted cells. Finally, we showed the colocalization of the WNV envelope protein and cholera toxin B, which is internalized in a lipid raft-dependent pathway, in microdomain clusters at the plasma membrane. These data suggest that WNV utilizes lipid rafts during initial stages of internalization and that the lipid rafts may contain a factor(s) that may enhance WNV endocytosis. PMID- 18385234 TI - Attenuation of V- or C-defective measles viruses: infection control by the inflammatory and interferon responses of rhesus monkeys. AB - Patients recruited in virus-based cancer clinical trials and immunocompromised individuals in need of vaccination would profit from viral strains with defined attenuation mechanisms. We generated measles virus (MV) strains defective for the expression of either the V protein, a modulator of the innate immune response, or the C protein, which has multiple functions. The virulence of these strains was compared with that of the parental wild-type MV in a natural host, Macaca mulatta. Skin rash, viremia, and the strength of the innate and adaptive immune responses were characterized in groups of six animals. Replication of V- or C protein-defective viruses was short-lived and reached lower levels in peripheral blood mononuclear cells and lymphatic organs compared to the wild-type virus; none of the mutants reverted to the wild type. The neutralizing antibody titers and MV-specific T-cell responses were equivalent in monkeys infected with the viral strains tested, documenting strong adaptive immune responses. In contrast, the inflammatory response was better controlled by wild-type MV, as revealed by inhibition of interleukin-6 and tumor necrosis factor alpha transcription. The interferon response was also better controlled by the wild-type virus than by the defective viruses. Since V- and C-defective MVs induce strong adaptive immune responses while spreading less efficiently, they may be developed as vaccines for immunocompromised individuals. Moreover, MV unable to interact with single innate immunity proteins may be developed for preferential replication in tumors with specific contexts of vulnerability. PMID- 18385235 TI - Insulin-like growth factor II mRNA binding protein 1 associates with Gag protein of human immunodeficiency virus type 1, and its overexpression affects virus assembly. AB - The assembly of human immunodeficiency virus type 1 (HIV-1) particles is driven by viral Gag protein. This function of Gag not only benefits from its self multimerization property but also depends on its interaction with a number of cellular factors such as TSG101 and ALIX/AIP1 that promote virus budding and release from cell surfaces. However, interaction with Gag also allows some cellular factors such as APOBEC3G and Trim5alpha to access viral replication machinery and block viral replication. In this study, we report a new HIV-1 Gag binding factor named insulin-like growth factor II mRNA binding protein 1 (IMP1). Gag-IMP1 interaction requires the second zinc finger of the nucleocapsid (NC) domain of Gag and the KH3 and KH4 domains of IMP1. A fourfold reduction of HIV-1 infectivity was seen with overexpression of the wild-type IMP1 and its mutant that is able to interact with Gag but not with overexpression of IMP1 mutants exhibiting Gag-binding deficiency. The decreased viral infectivity was further shown as a result of diminished viral RNA packaging, abrogated Gag processing on the cellular membranes, and impeded maturation of virus particles. Together, these results demonstrate that IMP1 interacts with HIV-1 Gag protein and is able to block the formation of infectious HIV-1 particles. PMID- 18385236 TI - Structural basis for the receptor binding specificity of Norwalk virus. AB - Noroviruses are positive-sense, single-stranded RNA viruses that cause acute gastroenteritis. They recognize human histo-blood group antigens as receptors in a strain-specific manner. The structures presented here were analyzed in order to elucidate the structural basis for differences in ligand recognition of noroviruses from different genogroups, the prototypic Norwalk virus (NV; GI-1) and VA387 (GII-4), which recognize the same A antigen but differ in that NV is unable to bind to the B antigen. Two forms of the receptor-binding domain of the norovirus coat protein, the P domain and the P polypeptide, that were previously shown to differ in receptor binding and P-particle formation properties were studied. Comparison of the structures of the NV P domain with and without A trisaccharide and the NV P polypeptide revealed no major ligand-induced changes. The 2.3-A cocrystal structure reveals that the A trisaccharide binds to the NV P domain through interactions with the residues Ser377, Asp327, His329, and Ser380 in a mode distinct from that previously reported for the VA387 P-domain-A trisaccharide complex. Mutational analyses confirm the importance of these residues in NV P-particle binding to native A antigen. The alpha-GalNAc residue unique to the A trisaccharide is buried deeply in the NV binding pocket, unlike in the structures of A and B trisaccharides bound to VA387 P domain, where the alpha-fucose residue forms the most protein contacts. The A-trisaccharide binding mode seen in the NV P domain complex cannot be sterically accommodated in the VA387 P domain. PMID- 18385238 TI - Tumor antigen LRRC15 impedes adenoviral infection: implications for virus-based cancer therapy. AB - Adenoviruses for gene or oncolytic therapy are under development. Notable among these strategies is adenoviral delivery of the tumor suppressor p53. Since all therapeutics have limitations in certain settings, we have undertaken retroviral suppressor screens to identify genes conferring resistance to adenovirus delivered p53. These studies identified the tumor antigen LRRC15, which is frequently overexpressed in multiple tumor types, as a repressor of cell death due to adenoviral p53. LRRC15, however, does not impede p53 function per se but impedes adenoviral infection. Specifically, LRRC15 causes redistribution of the coxsackievirus-adenovirus receptor away from the cell surface. This effect is manifested in less adenoviral binding to the surfaces of LRRC15-expressing cells. This discovery, therefore, not only is important for understanding adenoviral biology but also has potentially important implications for adenovirus-based anticancer therapeutics. PMID- 18385237 TI - Intrinsic structural disorder in adenovirus E1A: a viral molecular hub linking multiple diverse processes. PMID- 18385239 TI - Identification of a highly conserved, functional nuclear localization signal within the N-terminal region of herpes simplex virus type 1 VP1-2 tegument protein. AB - VP1-2 is a large structural protein assembled into the tegument compartment of the virion, conserved across the herpesviridae, and essential for virus replication. In herpes simplex virus (HSV) and pseudorabies virus, VP1-2 is tightly associated with the capsid. Studies of its assembly and function remain incomplete, although recent data indicate that in HSV, VP1-2 is recruited onto capsids in the nucleus, with this being required for subsequent recruitment of additional structural proteins. Here we have developed an antibody to characterize VP1-2 localization, observing the protein in both cytoplasmic and nuclear compartments, frequently in clusters in both locations. Within the nucleus, a subpopulation of VP1-2 colocalized with VP26 and VP5, though VP1-2 positive foci devoid of these components were observed. We note a highly conserved basic motif adjacent to the previously identified N-terminal ubiquitin hydrolase domain (DUB). The DUB domain in isolation exhibited no specific localization, but when extended to include the adjacent motif, it efficiently accumulated in the nucleus. Transfer of the isolated motif to a test protein, beta-galactosidase, conferred specific nuclear localization. Substitution of a single amino acid within the motif abolished the nuclear localization function. Deletion of the motif from intact VP1-2 abrogated its nuclear localization. Moreover, in a functional assay examining the ability of VP1-2 to complement growth of a VP1-2-ve mutant, deletion of the nuclear localization signal abolished complementation. The nuclear localization signal may be involved in transport of VP1-2 early in infection or to late assembly sites within the nucleus or, considering the potential existence of VP1-2 cleavage products, in selective localization of subdomains to different compartments. PMID- 18385240 TI - A novel mutation in murine hepatitis virus nsp5, the viral 3C-like proteinase, causes temperature-sensitive defects in viral growth and protein processing. AB - Sequencing and reversion analysis of murine hepatitis virus (MHV) temperature sensitive (ts) viruses has identified putative ts mutations in the replicase nonstructural proteins (nsp's) of these coronaviruses. In this study, reverse transcriptase PCR sequencing of the RNA genome of an isolate of the MHV ts virus Alb ts6, referred to as Alb/ts/nsp5/V148A, identified a putative ts mutation in nsp5 (T10651C, Val148Ala), the viral 3C-like proteinase (3CLpro). The introduction of the T10651C mutation into the infectious MHV clone resulted in the recovery of a mutant virus, the nsp5/V148A virus, that demonstrated reduced growth and nsp5 proteinase activity identical to that of Alb/ts/nsp5/V148A at the nonpermissive temperature. Sequence analysis of 40 degrees C revertants of Alb/ts/nsp5/V148A identified primary reversion to Ala148Val in nsp5, as well as two independent second-site mutations resulting in Ser133Asn and His134Tyr substitutions in nsp5. The introduction of the Ser133Asn or His134Tyr substitution into the cloned nsp5/V148A mutant virus background resulted in the recovery of viruses with increased growth fitness and the partial restoration of nsp5 activity at the nonpermissive temperature. Modeling of the nsp5 structure of Alb/ts/nsp5/V148A predicted that the Val148Ala mutation alters residue 148 interactions with residues of the substrate binding S1 subsite of the nsp5 active site cavity. This study identifies novel residues in nsp5 that may be important for regulating substrate specificity and nsp5 proteinase activity. PMID- 18385241 TI - Multimerization of tegument protein pp28 within the assembly compartment is required for cytoplasmic envelopment of human cytomegalovirus. AB - Human cytomegalovirus (HCMV) UL99-encoded pp28 is an essential tegument protein required for envelopment and production of infectious virus. Nonenveloped virions accumulate in the cytoplasm of cells infected with recombinant viruses with the UL99 gene deleted. Previous results have suggested that a key function of pp28 in the envelopment of infectious HCMV is expressed after the protein localizes in the assembly compartment (AC). In this study, we investigated the potential role of pp28 multimerization in the envelopment of the infectious virion. Our results indicated that pp28 multimerized during viral infection and that interacting domains responsible for self-interaction were localized in the amino terminus of the protein (amino acids [aa] 1 to 43). The results from transient-expression and/or infection assays indicated that the self-interaction took place in the AC. A mutant pp28 molecule containing only the first 35 aa failed to accumulate in the AC, did not interact with pp28 in the AC, and could not support virus replication. In contrast, the first 50 aa of pp28 was sufficient for the self interaction within the AC and the assembly of infectious virus. Recombinant viruses encoding an in-frame deletion of aa 26 to 33 of pp28 were replication competent, whereas infectious virus was not recovered from HCMV BACs lacking aa 26 to 43. These findings suggested that the accumulation of pp28 was a prerequisite for multimerization of pp28 within the AC and that pp28 multimerization in the AC represented an essential step in the envelopment and production of infectious virions. PMID- 18385242 TI - A point mutation leading to hepatitis C virus escape from neutralization by a monoclonal antibody to a conserved conformational epitope. AB - A challenge in hepatitis C virus (HCV) vaccine development is defining conserved protective epitopes. A cluster of these epitopes comprises an immunodominant domain on the E2 glycoprotein, designated domain B. CBH-2 is a neutralizing human monoclonal antibody to a domain B epitope that is highly conserved. Alanine scanning demonstrated that the epitope involves residues G523, G530, and D535 that are also contact residues for E2 binding to CD81, a coreceptor required for virus entry into cells. However, another residue, located at position 431 and thus at a considerable distance in the linear sequence of E2, also contributes to the CBH-2 epitope. A single amino acid substitution at this residue results in escape from CBH-2-mediated neutralization in a genotype 1a virus. These results highlight the challenges inherent in developing HCV vaccines and show that an effective vaccine must induce antibodies to both conserved and more invariant epitopes to minimize virus escape. PMID- 18385243 TI - The affinity of EBNA1 for its origin of DNA synthesis is a determinant of the origin's replicative efficiency. AB - Epstein-Barr virus (EBV) replicates its genome as a licensed plasmid in latently infected cells. Although replication of this plasmid is essential for EBV latent infection, its synthesis still fails for 16% of the templates in S phase. In order to understand these failures, we sought to determine whether the affinity of the initiator protein (EBNA1) for its binding sites in the origin affects the efficiency of plasmid replication. We have answered this question by using several engineered origins modeled upon the arrangement of EBNA1-binding sites found in DS, the major plasmid origin of EBV. The human TRF2 protein also binds to half-sites in DS and increases EBNA1's affinity for its own sites; we therefore also tested origin efficiency in the presence or absence of these sites. We have found that if TRF2-half-binding sites are present, the efficiency of supporting the initiation of DNA synthesis and of establishing a plasmid bearing that origin directly correlates with the affinity of EBNA1 for that origin. Moreover, the presence of TRF2-half-binding sites also increases the average level of EBNA1 and ORC2 bound to those origins in vivo, as measured by chromatin immunoprecipitation. Lastly, we have created an origin of DNA synthesis from high-affinity EBNA1-binding sites and TRF2-half-binding sites that functions severalfold more efficiently than does DS. This finding indicates that EBV has selected a submaximally efficient origin of DNA synthesis for the latent phase of its life cycle. This enhanced origin could be used practically in human gene vectors to improve their efficiency in therapy and basic research. PMID- 18385244 TI - Human immunodeficiency virus type 1 Vpr binds to the N lobe of the Wee1 kinase domain and enhances kinase activity for CDC2. AB - Human immunodeficiency virus type 1 Vpr is a virion-associated accessory protein that has multiple activities within an infected cell. One of the most dramatic effects of Vpr is the induction of cell cycle arrest at the G(2)/M boundary, followed by apoptosis. This effect has implications for CD4(+) cell loss in AIDS. In normal cell cycle regulation, Wee1, a key regulator for G(2)-M progression, phosphorylates Tyr15 on Cdc2 and thereby blocks the progression of cells into M phase. We demonstrate that Vpr physically interacts with Wee1 at the N lobe of the kinase domain analogous to that present in other kinases. This interaction with Vpr enhances Wee1 kinase activity for Cdc2. Overexpression of Wee1 kinase deficient mutants competes for Vpr-mediated cell cycle arrest, and deletion of the region of Wee1 that binds Vpr abrogates that competition. However, the Vpr mutants I74P and I81P, which fail to induce G(2) arrest, can bind to and increase the kinase activity of Wee1 to the same extent as wild-type Vpr. Therefore, we conclude that the binding of Vpr to Wee1 is not sufficient for Vpr to activate the G(2) checkpoint, and it may reflect an independent function of Vpr. PMID- 18385245 TI - Transformation by bovine papillomavirus type 1 E6 requires paxillin. AB - Papillomavirus E6 proteins are adapters that change the function of cellular regulatory proteins. The bovine papillomavirus type 1 E6 (BE6) binds to LXXLL peptide sequences termed LD motifs (consensus sequence LDXLLXXL) on the cellular protein paxillin that is a substrate of Src and focal adhesion kinases. Anchorage independent transformation induced by BE6 required both paxillin and BE6-binding LD motifs on paxillin but was independent of the major tyrosine phosphorylation sites of paxillin. The essential role of paxillin in transformation by BE6 highlights the role of paxillin in the transduction of cellular signals that result in anchorage-independent cell proliferation. PMID- 18385246 TI - Different mutational pathways to CXCR4 coreceptor switch of CCR5-using simian human immunodeficiency virus. AB - We report here a second case of coreceptor switch in R5 simian-human immunodeficiency virus SF162P3N (SHIV(SF162P3N))-infected macaque CA28, supporting the use of this experimental system to examine factors that drive the change in coreceptor preference in vivo. Virus recovered from CA28 plasma (SHIV(CA28NP)) used both CCR5 and CXCR4 for entry, but the virus recovered from lymph node (SHIV(CA28NL)) used CXCR4 almost exclusively. Sequence and functional analyses showed that mutations in the V3 loop that conferred CXCR4 usage in macaque CA28 differed from those described in the previously reported case, demonstrating divergent mutational pathways for change in the coreceptor preference of the R5 SHIV(SF162P3N) isolate in vivo. PMID- 18385247 TI - Insertion of the two cleavage sites of the respiratory syncytial virus fusion protein in Sendai virus fusion protein leads to enhanced cell-cell fusion and a decreased dependency on the HN attachment protein for activity. AB - Cell entry by paramyxoviruses requires fusion of the viral envelope with the target cell membrane. Fusion is mediated by the viral fusion (F) glycoprotein and usually requires the aid of the attachment glycoprotein (G, H or HN, depending on the virus). Human respiratory syncytial virus F protein (F(RSV)) is able to mediate membrane fusion in the absence of the attachment G protein and is unique in possessing two multibasic furin cleavage sites, separated by a region of 27 amino acids (pep27). Cleavage at both sites is required for cell-cell fusion. We have investigated the significance of the two cleavage sites and pep27 in the context of Sendai virus F protein (F(SeV)), which possesses a single monobasic cleavage site and requires both coexpression of the HN attachment protein and trypsin in order to fuse cells. Inclusion of both F(RSV) cleavage sites in F(SeV) resulted in a dramatic increase in cell-cell fusion activity in the presence of HN. Furthermore, chimeric F(SeV) mutants containing both F(RSV) cleavage sites demonstrated cell-cell fusion in the absence of HN. The presence of two multibasic cleavage sites may therefore represent a strategy to regulate activation of a paramyxovirus F protein for cell-cell fusion in the absence of an attachment protein. PMID- 18385248 TI - Recombinant vesicular stomatitis virus vector mediates postexposure protection against Sudan Ebola hemorrhagic fever in nonhuman primates. AB - Recombinant vesicular stomatitis virus (VSV) vectors expressing homologous filoviral glycoproteins can completely protect rhesus monkeys against Marburg virus when administered after exposure and can partially protect macaques after challenge with Zaire ebolavirus. Here, we administered a VSV vector expressing the Sudan ebolavirus (SEBOV) glycoprotein to four rhesus macaques shortly after exposure to SEBOV. All four animals survived SEBOV challenge, while a control animal that received a nonspecific vector developed fulminant SEBOV hemorrhagic fever and succumbed. This is the first demonstration of complete postexposure protection against an Ebola virus in nonhuman primates and provides further evidence that postexposure vaccination may have utility in treating exposures to filoviruses. PMID- 18385249 TI - Demyelinating and nondemyelinating strains of mouse hepatitis virus differ in their neural cell tropism. AB - Some strains of mouse hepatitis virus (MHV) can induce chronic inflammatory demyelination in mice that mimics certain pathological features of multiple sclerosis. We have examined neural cell tropism of demyelinating and nondemyelinating strains of MHV in order to determine whether central nervous system (CNS) cell tropism plays a role in demyelination. Previous studies demonstrated that recombinant MHV strains, isogenic other than for the spike gene, differ in the extent of neurovirulence and the ability to induce demyelination. Here we demonstrate that these strains also differ in their abilities to infect a particular cell type(s) in the brain. Furthermore, there is a correlation between the differential localization of viral antigen in spinal cord gray matter and that in white matter during acute infection and the ability to induce demyelination later on. Viral antigen from demyelinating strains is detected initially in both gray and white matter, with subsequent localization to white matter of the spinal cord, whereas viral antigen localization of nondemyelinating strains is restricted mainly to gray matter. This observation suggests that the localization of viral antigen to white matter during the acute stage of infection is essential for the induction of chronic demyelination. Overall, these observations suggest that isogenic demyelinating and nondemyelinating strains of MHV, differing in the spike protein expressed, infect neurons and glial cells in different proportions and that differential tropism to a particular CNS cell type may play a significant role in mediating the onset and mechanisms of demyelination. PMID- 18385250 TI - Endoplasmic reticulum chaperones are involved in the morphogenesis of rotavirus infectious particles. AB - The final assembly of rotavirus particles takes place in the endoplasmic reticulum (ER). In this work, we evaluated by RNA interference the relevance to rotavirus assembly and infectivity of grp78, protein disulfide isomerase (PDI), grp94, calnexin, calreticulin, and ERp57, members of the two ER folding systems described herein. Silencing the expression of grp94 and Erp57 had no effect on rotavirus infectivity, while knocking down the expression of any of the other four chaperons caused a reduction in the yield of infectious virus of about 50%. In grp78-silenced cells, the maturation of the oligosaccharide chains of NSP4 was retarded. In cells with reduced levels of calnexin, the oxidative folding of VP7 was impaired and the trimming of NSP4 was accelerated, and in calreticulin silenced cells, the formation of disulfide bonds of VP7 was also accelerated. The knockdown of PDI impaired the formation and/or rearrangement of the VP7 disulfide bonds. All these conditions also affected the correct assembly of virus particles, since compared with virions from control cells, they showed an altered susceptibility to EGTA and heat treatments, a decreased specific infectivity, and a diminished reactivity to VP7 with monoclonal antibody M60, which recognizes only this protein when its disulfide bonds have been correctly formed. In the case of grp78-silenced cells, the virus produced bound less efficiently to MA104 cells than virus obtained from control cells. All these results suggest that these chaperones are involved in the quality control of rotavirus morphogenesis. The complexity of the steps of rotavirus assembly that occur in the ER provide a useful model for studying the organization and operation of the complex network of chaperones involved in maintaining the quality control of this organelle. PMID- 18385251 TI - Comprehensive immunological evaluation reveals surprisingly few differences between elite controller and progressor Mamu-B*17-positive simian immunodeficiency virus-infected rhesus macaques. AB - The association between particular major histocompatibility complex class I (MHC I) alleles and control of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) replication implies that certain CD8(+) T-lymphocyte (CD8-TL) responses are better able than others to control viral replication in vivo. However, possession of favorable alleles does not guarantee improved prognosis or viral control. In rhesus macaques, the MHC-I allele Mamu-B*17 is correlated with reduced viremia and is overrepresented in macaques that control SIVmac239, termed elite controllers (ECs). However, there is so far no mechanistic explanation for this phenomenon. Here we show that the chronic-phase Mamu-B*17-restricted repertoire is focused primarily against just five epitopes VifHW8, EnvFW9, NefIW9, NefMW9, and env(ARF)cRW9-in both ECs and progressors. Interestingly, Mamu-B*17-restricted CD8-TL do not target epitopes in Gag. CD8-TL escape variation occurred in all targeted Mamu-B*17-restricted epitopes. However, recognition of escape variant peptides was commonly observed in both ECs and progressors. Wild-type sequences in the VifHW8 epitope tended to be conserved in ECs, but there was no evidence that this enhances viral control. In fact, no consistent differences were detected between ECs and progressors in any measured parameter. Our data suggest that the narrowly focused Mamu-B*17-restricted repertoire suppresses virus replication and drives viral evolution. It is, however, insufficient in the majority of individuals that express the "protective" Mamu-B*17 molecule. Most importantly, our data indicate that the important differences between Mamu-B*17-positive ECs and progressors are not readily discernible using standard assays to measure immune responses. PMID- 18385252 TI - Characterization of the follicular dendritic cell reservoir of human immunodeficiency virus type 1. AB - Throughout the natural course of human immunodeficiency virus (HIV) infection, follicular dendritic cells (FDCs) trap and retain large quantities of particle associated HIV RNA in the follicles of secondary lymphoid tissue. We have previously found that murine FDCs in vivo could maintain trapped virus particles in an infectious state for at least 9 months. Here we sought to determine whether human FDCs serve as an HIV reservoir, based on the criteria that virus therein must be replication competent, genetically diverse, and archival in nature. We tested our hypothesis using postmortem cells and tissues obtained from three HIV infected subjects and antemortem blood samples obtained from one of these subjects. Replication competence was determined using coculture, while genetic diversity and the archival nature of virus were established using phylogenetic and population genetics methods. We found that FDC-trapped virus was replication competent and demonstrated greater genetic diversity than that of virus found in most other tissues and cells. Antiretrovirus-resistant variants that were not present elsewhere were also detected on FDCs. Furthermore, genetic similarity was observed between FDC-trapped HIV and viral species recovered from peripheral blood mononuclear cells obtained 21 and 22 months antemortem, but was not present in samples obtained 4 and 18 months prior to the patient's death, indicating that FDCs can archive HIV. These data indicate that FDCs represent a significant reservoir of infectious and diverse HIV, thereby providing a mechanism for viral persistence for months to years. PMID- 18385253 TI - A direct comparison of human papillomavirus type 16 L1 particles reveals a lower immunogenicity of capsomeres than viruslike particles with respect to the induced antibody response. AB - Capsomeres are considered to be an alternative to viruslike particle (VLP)-based vaccines as they can be produced in prokaryotic expression systems. So far, no detailed side-by-side comparison of VLPs and capsomeres has been performed. In the present study, we immunized mice with insect cell-derived human papillomavirus type 16 VLPs and capsomeres. VLPs induced consistently higher antibody titers than capsomeres but the two forms induced similar CD8 T-cell responses after subcutaneous, intranasal, and oral immunization, and at least 20 to 40 times more L1 in the form of capsomeres than in the form of VLPs was needed to achieve comparable antibody responses. These results were confirmed by DNA immunization. The lower immunogenicity of capsomeres was independent of the isotype switch, as it was also observed for the early immunoglobulin M responses. Although there were differences in the display of surface epitopes between the L1 particles, these did not contribute significantly to the differences in the immune responses. capsomeres were less immunogenic than VLPs in Toll-like receptor 4 (TLR4)-deficient mice, suggesting that the lower immunogenicity is not due to a failure of capsomeres to trigger TLR4. We observed better correlation between antibody results from enzyme-linked immunosorbent assays and neutralization assays for sera from VLP-immunized mice than for sera from capsomere-immunized mice, suggesting qualitative differences between VLPs and capsomeres. We also showed that the lower immunogenicity of capsomeres could be compensated by the use of an adjuvant system containing MPL. Taken together, these results suggest that, presumably because of the lower degree of complexity of the antigen organization, capsomeres are significantly less immunogenic than VLPs with respect to the humoral immune response and that this characteristic should be considered in the design of putative capsomere-based prophylactic vaccines. PMID- 18385254 TI - Natural resistance of human immunodeficiency virus type 1 to the CD4bs antibody b12 conferred by a glycan and an arginine residue close to the CD4 binding loop. AB - The human monoclonal antibody b12 recognizes a conserved epitope on gp120 that overlaps the CD4 binding site. b12 has neutralizing activity against diverse human immunodeficiency virus type 1 (HIV-1) strains. However, we recently reported that b12 sensitivity of HIV-1 envelopes amplified from patient tissues without culture varied considerably. For two subjects, there was clear modulation of b12 sensitivity, with lymph node-derived envelopes being essentially resistant while those from brain tissue were sensitive. Here, we have mapped envelope determinants of b12 resistance by constructing chimeric envelopes from resistant and sensitive envelopes derived from lymph node and brain tissue, respectively. Residues on the N-terminal flank of the CD4 binding loop conferred partial resistance. However, a potential glycosylation site at residue N386 completely modulated b12 resistance but required the presence of an arginine at residue 373. Moreover, the introduction of R373 into b12-sensitive NL4.3 and AD8 envelopes, which carry N386, also conferred b12 resistance. Molecular modeling suggests that R373 and the glycan at N386 may combine to sterically exclude the benzene ring of b12 W100 from entering a proximal pocket. In summary, we identify residues on either side of the CD4 binding loop that contribute to b12 resistance in immune tissue in vivo. Our data have relevance for the design of vaccines that aim to elicit neutralizing antibodies. PMID- 18385256 TI - Critical numbers of neural crest cells are required in the pathways from the neural tube to the foregut to ensure complete enteric nervous system formation. AB - The enteric nervous system (ENS) is mainly derived from vagal neural crest cells (NCC) that arise at the level of somites 1-7. To understand how the size and composition of the NCC progenitor pool affects ENS development, we reduced the number of NCC by ablating the neural tube adjacent to somites 3-6 to produce aganglionic gut. We then back-transplanted various somite lengths of quail neural tube into the ablated region to determine the 'tipping point', whereby sufficient progenitors were available for complete ENS formation. The addition of one somite length of either vagal, sacral or trunk neural tube into embryos that had the neural tube ablated adjacent to somites 3-6, resulted in ENS formation along the entire gut. Although these additional cells contributed to the progenitor pool, the quail NCC from different axial levels retained their intrinsic identities with respect to their ability to form the ENS; vagal NCC formed most of the ENS, sacral NCC contributed a limited number of ENS cells, and trunk NCC did not contribute to the ENS. As one somite length of vagal NCC was found to comprise almost the entire ENS, we ablated all of the vagal neural crest and back transplanted one somite length of vagal neural tube from the level of somite 1 or somite 3 into the vagal region at the position of somite 3. NCC from somite 3 formed the ENS along the entire gut, whereas NCC from somite 1 did not. Intrinsic differences, such as an increased capacity for proliferation, as demonstrated in vitro and in vivo, appear to underlie the ability of somite 3 NCC to form the entire ENS. PMID- 18385255 TI - Alphaherpesviruses and chemokines: pas de deux not yet brought to perfection. PMID- 18385258 TI - Dynamic regulation of the expression of neurotrophin receptors by Runx3. AB - Sensory neurons in the dorsal root ganglion (DRG) specifically project axons to central and peripheral targets according to their sensory modality. However, the molecular mechanisms that govern sensory neuron differentiation and the axonal projections remain unclear. The Runt-related transcription factors, Runx1 and Runx3, are expressed in DRG neuronal subpopulations, suggesting that they might regulate the cell specification and the trajectories of specific axons. Here, we show that parvalbumin-positive DRG neurons fail to differentiate from the onset in Runx3(-/-) mice. By contrast, TrkC-positive DRG neurons differentiate normally at embryonic day (E) 11.5, but disappear by E13.5 in Runx3(-/-) mice. Subsequently, TrkC-positive DRG neurons reappear but in smaller numbers than in the wild type. In Runx3(-/-) mice, central axons of the TrkC-positive DRG neurons project to the dorsal spinal cord but not to the ventral and intermediate spinal cord, whereas the peripheral axons project to skin but not to muscle. These results suggest that Runx3 controls the acquisition of distinct proprioceptive DRG neuron identities, and that TrkC-positive DRG neurons consist of two subpopulations: Runx3-dependent early-appearing proprioceptive neurons that project to the ventral and intermediate spinal cord and muscle; and Runx3 independent late-appearing cutaneous neurons that project to the dorsal spinal cord and skin. Moreover, we show that the number of TrkA-positive DRG neurons is reduced in Runx3(-/-) mice, as compared with the wild type. These results suggest that Runx3 positively regulates the expression of TrkC and TrkA in DRG neurons. PMID- 18385257 TI - Tbx2b is required for the development of the parapineal organ. AB - Structural differences between the left and right sides of the brain exist throughout the vertebrate lineage. By studying the zebrafish pineal complex, which exhibits notable asymmetries, both the genes and the cell movements that result in left-right differences can be characterized. The pineal complex consists of the midline pineal organ and the left-sided parapineal organ. The parapineal is responsible for instructing the asymmetric architecture of the bilateral habenulae, the brain nuclei that flank the pineal complex. Using in vivo time-lapse confocal microscopy, we find that the cells that form the parapineal organ migrate as a cluster of cells from the pineal complex anlage to the left side of the brain. In a screen for mutations that disrupted brain laterality, we identified a nonsense mutation in the T-box2b (tbx2b) gene, which encodes a transcription factor expressed in the pineal complex anlage. The tbx2b mutant makes fewer parapineal cells, and they remain as individuals near the midline rather than migrating leftward as a group. The reduced number and incorrect placement of parapineal cells result in symmetric development of the adjacent habenular nuclei. We conclude that tbx2b functions to specify the correct number of parapineal cells and to regulate their asymmetric migration. PMID- 18385259 TI - Genetic aspects of female reproduction. AB - BACKGROUND: Sexual reproduction provides the means for preserving genetic identity and in turn, genetic variability may affect the ability to reproduce. This review aims to summarize current research on genetic diagnosis and genetic causes of reproductive disorders. METHODS: Searches were done by subject in Medline and other databases, and each subject summary was presented to the Workshop Group and omissions or disagreements were resolved by discussion. RESULTS: Single-gene defects are most likely to be found among patients with hypogonadotropic hypogonadism, which may be due to defects in the KAL genes or the gonadotrophin-releasing hormone receptor genes. With premature ovarian failure there is an increased risk of having a premutation of the Fragile X syndrome gene. Complex genetic inheritance may explain the variable familial links in polycystic ovary syndrome and endometriosis, but no definitive genetic pathways are as yet known. With recurrent miscarriage, genetic defects causing thrombophilias are 2-fold more likely. Chromosome abnormalities account for approximately 60% of all spontaneous abortions, and the most common type, trisomy, is closely associated with advanced maternal age. Three percent of couples have a balanced chromosome abnormality, but live birth rates are better with natural conception than with preimplantation genetic diagnosis. CONCLUSIONS: Understanding of the methods used for genetic diagnosis and research is becoming a standard requirement for the clinical practice of reproductive medicine. PMID- 18385260 TI - Targeting the vascular endothelial growth factor system to prevent ovarian hyperstimulation syndrome. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) typically occurs when ovaries are primed with FSH/LH and subsequently exposed to hCG. The ultimate pathophysiological step underlying this clinical picture is increased vascular permeability (VP). METHODS: A search of the literature was carried out using PubMed and the authors' files. RESULTS: In rodents and humans, the expression of vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR-2) mRNA increases during ovarian stimulation. With the administration of hCG, the expression of each rises to a maximum. Expression of VEGF/VEGFR-2 mRNAs correlates with enhanced VP, with both peaking 48 h following an injection of hCG. Immunohistochemistry shows the presence of VEGF and VEGFR-2 proteins in the granulosa-lutein and endothelial cells of the entire corpus luteum. Increased VP may be mediated through adhesion molecules such as VE-cadherin, which is involved in the loosening of endothelial intercellular junctions. These findings regarding the pathophysiology of OHSS suggest that the syndrome can be prevented by inducing ovulation with LH or GnRH analogues, which prevent VEGF overexpression. Also, co-administration of a dopamine agonist inhibits phosphorylation of the receptor VEGFR-2. In a trial of 69 oocyte donors, the incidence of moderate OHSS was 20% with the dopamine agonist cabergoline and 44% with a placebo (P = 0.04). CONCLUSIONS: The pathophysiological mechanisms involved in OHSS suggest potential preventive approaches, but larger trials are necessary for evaluating the efficacy and safety of the pharmaco-prevention of OHSS. PMID- 18385261 TI - Examination of the effect of procalcitonin on human leucocytes and the porcine isolated coronary artery. AB - BACKGROUND: The aim of this study was to investigate the effects of procalcitonin on the lipopolysaccharide (LPS)-induced changes in human leucocytes and porcine isolated coronary artery. METHODS: Using flow cytometry, changes in forward scatter and intracellular calcium in human neutrophils and monocytes were determined after exposure to procalcitonin, calcitonin gene-related peptide (CGRP), LPS, and the known chemoattractants formylated methionine-leucine phenylalanine (fMLP) and interleukin-8 (IL-8). In porcine isolated coronary artery, the effects of procalcitonin were evaluated using the contractile function change and the release of TNFalpha. RESULTS: In human neutrophils and monocytes, procalcitonin (100 nM), but not CGRP, increased forward scatter and the expression of surface markers (CD16 and CD14, respectively) in a similar manner to 10 microg ml(-1) LPS. Procalcitonin, but not CGRP, also increased the proportion of cells exhibiting an increase in intracellular calcium ions similar to that produced by fMLP and IL-8. Acute exposure of the coronary artery to procalcitonin produced a small, endothelium-independent relaxation (approximately 15% of constrictor tone), but failed to modify subsequent relaxations to CGRP. After 16 h exposure, procalcitonin (100 nM) increased TNFalpha release from the coronary artery equivalent to 70% of that produced by LPS, but did not modify the inhibitory effect of LPS (100 microg ml(-1)) on contractile responses. CONCLUSIONS: Procalcitonin has a proinflammatory effect on human leucocytes and porcine coronary artery, but it is not capable of modulating LPS-induced changes in vascular responsiveness in vitro. PMID- 18385262 TI - Plasma concentrations and sedation scores after nebulized and intranasal midazolam in healthy volunteers. AB - BACKGROUND: An efficacious, reliable, and non-invasive route of administration for midazolam, a drug used for sedation and pre-anaesthetic medication, would have obvious advantages. This study compares two potential methods of administering midazolam by the nasal and nebulized routes. METHODS: Midazolam (0.2 mg kg(-1)) was given by both nebulizer and nasally by liquid instillation to 10 healthy volunteers in a randomized, double-blind crossover study. Plasma concentrations of midazolam, Ramsay sedation scores, visual analogue scores, critical flicker fusion frequency, and parameters of cardiovascular and respiratory function were measured over 60 min and summarized using 'area under the curve'. RESULTS: Nasal instillation caused more sedation than nebulized administration. This was demonstrated by higher Ramsay sedation scores (P=0.005), lower visual analogue scores (P<0.001), and lower critical flicker fusion frequency (P<0.02). Nasal instillation was associated with higher plasma concentrations of midazolam (P<0.001). Unpleasant symptoms were recorded by six volunteers in the intranasal and one in the nebulized group (P=0.06). CONCLUSIONS: There was some evidence that midazolam caused less discomfort when given by nebulizer compared with intranasally. Comparative bioavailability of midazolam, estimated by the ratio (nebulized:nasal) of area under the 60 min plasma concentration curve, was 1:2.9. A higher dose may need to be administered for adequate pre-anaesthetic medication when midazolam is given by nebulizer. PMID- 18385263 TI - Signs of myocardial ischaemia after injection of oxytocin: a randomized double blind comparison of oxytocin and methylergometrine during Caesarean section. AB - BACKGROUND: ECG changes, similar to those seen during myocardial ischaemia, together with symptoms of chest pain, are common during Caesarean section (CS). We hypothesized that oxytocin administration has cardiovascular effects leading to these symptoms and ECG changes. METHODS: Forty women undergoing elective CS under spinal anaesthesia were given an i.v. bolus of either 10 IU of oxytocin (Group OXY-CS, n=20) or 0.2 mg of methylergometrine (Group MET-CS, n=20), in a double-blind, randomized fashion after delivery. Ten healthy, non-pregnant, non anaesthetized women were used as normal controls (Group OXY-NC, n=10) and were given 10 IU of oxytocin i.v. Twelve-lead ECG, on-line, computerized vectorcardiography (VCG), and invasive arterial pressure were recorded. RESULTS: Oxytocin produced a significant increase in heart rate, +28 (SD 4) and +52 (3) beats min(-1) [mean (SEM); P<0.001], decreases in mean arterial pressure, -33 (2) and -30 (3) mm Hg (P<0.001), and increases in the spatial ST-change vector magnitude (STC-VM), +77 (12) and +114 (8) microV (P<0.001), in CS patients and controls, respectively. Symptoms of chest pain and subjective discomfort were simultaneously present. Methylergometrine produced mild hypertension and no significant ECG changes. CONCLUSIONS: Oxytocin administered as an i.v. bolus of 10 IU induces chest pain, transient profound tachycardia, hypotension, and concomitant signs of myocardial ischaemia according to marked ECG and STC-VM changes. The effects are related to oxytocin administration and not to pregnancy, surgical procedure, delivery, or sympathetic block from spinal anaesthesia. PMID- 18385264 TI - Predicting death and readmission after intensive care discharge. AB - BACKGROUND: Despite initial recovery from critical illness, many patients deteriorate after discharge from the intensive care unit (ICU). We examined prospectively collected data in an attempt to identify patients at risk of readmission or death after intensive care discharge. METHODS: This was a secondary analysis of clinical audit data from patients discharged alive from a mixed medical and surgical (non-cardiac) ICU. RESULTS: Four hundred and seventy five patients (11.2%) died in hospital after discharge from the ICU. Increasing age, time in hospital before intensive care admission, Acute Physiology and Chronic Health Evaluation II (APACHE II) score, and discharge Therapeutic Intervention Scoring System (TISS) score were independent risk factors for death after intensive care discharge. Three hundred and eighty-five patients (8.8%) were readmitted to intensive care during the same hospital admission. Increasing age, time in hospital before intensive care, APACHE II score, and discharge to a high dependency unit were independent risk factors for readmission. One hundred and forty-three patients (3.3%) were readmitted within 48 h of intensive care discharge. APACHE II scores and discharge to a high dependency or other ICU were independent risk factors for early readmission. The overall discriminant ability of our models was moderate with only marginal benefit over the APACHE II scores alone. CONCLUSIONS: We identified risk factors associated with death and readmission to intensive care. It was not possible to produce a definitive model based on these risk factors for predicting death or readmission in an individual patient. PMID- 18385265 TI - Reversal of rocuronium-induced neuromuscular block with sugammadex is faster than reversal of cisatracurium-induced block with neostigmine. AB - BACKGROUND: Reversal of the residual effect of rocuronium or cisatracurium by neostigmine may be slow and associated with side-effects. This randomized, safety assessor-blinded study compared the efficacy of sugammadex, a selective relaxant binding agent for reversal of rocuronium-induced neuromuscular block, with that of neostigmine for reversal of cisatracurium-induced neuromuscular block. The safety of sugammadex and neostigmine was also evaluated. METHODS: Adult surgical patients (ASA class I-III) were randomized to sugammadex 2.0 mg kg(-1) for reversal of block induced by rocuronium 0.6 mg kg(-1), or neostigmine 50 microg kg(-1) for reversal of block induced by cisatracurium 0.15 mg kg(-1). Anaesthesia was induced and maintained using i.v. propofol and remifentanil, fentanyl, or sufentanil. Neuromuscular function was monitored using acceleromyography (TOF Watch SX). Sugammadex or neostigmine was administered at reappearance of T(2). The primary efficacy variable was time for recovery of the train-of-four (TOF) ratio to 0.9. RESULTS: Eighty-four patients were randomized, 73 of whom received sugammadex (n=34) or neostigmine (n=39). Time from start of administration of reversal agent to recovery of the TOF ratio to 0.9 was 4.7 times faster with sugammadex than with neostigmine (geometric mean=1.9 vs 9.0 min, P<0.0001). Reversal of block was sustained in all patients. There were no serious adverse effects from either reversal agent and no significant changes in any measure of safety, except for similar elevations in urinary N-acetyl glucosaminidase in both groups. CONCLUSIONS: Sugammadex 2.0 mg kg(-1) administered at reappearance of T(2) was significantly faster in reversing rocuronium-induced blockade than neostigmine was in reversing cisatracurium-induced block. PMID- 18385266 TI - Mouse model of type II Bartter's syndrome. I. Upregulation of thiazide-sensitive Na-Cl cotransport activity. AB - ROMK-deficient (Romk(-/-)) mice exhibit polyuria, natriuresis, and kaliuresis similar to individuals with type II Bartter's form of hyperprostaglandin E syndrome (HPS; antenatal Bartter's syndrome). In the present study, we utilized both metabolic and clearance studies to define the contributions of specific distal nephron segments to the renal salt wasting in these mice. The effects of furosemide, hydrochlorothiazide, and benzamil on urinary Na(+) and K(+) excretion in both wild-type (Romk(+/+)) and Romk(-/-) mice were used to assess and compare salt transport by the Na(+)-K(+)-2Cl(-) cotransporter (NKCC2)-expressing thick ascending limb (TAL), the Na(+)-Cl(-) cotransporter (NCC)-expressing distal convoluted tubule (DCT1/DCT2), and the epithelial Na(+) channel (ENaC)-expressing connecting segment (CNT) and collecting duct (CD), respectively. Whole kidney glomerular filtration rate was reduced by 47% in Romk(-/-) mice. Furosemide induced increments in the fractional excretion rate of Na(+) and K(+) and absolute excretion of Na(+) and K(+) were significantly blunted in Romk(-/-) mice, consistent with a major salt transport defect in the TAL. In contrast, hydrochlorothiazide produced an exaggerated natriuresis in Romk(-/-) mice, indicating upregulation of salt absorption by the DCT. Benzamil resulted in a similar increment in absolute Na excretion in both Romk(-/-) and Romk(+/+), indicating no significant upregulation of Na(+) transport by ENaC in ROMK null mice. Moreover, hydrochlorothiazide increased the fractional K(+) excretion rate in Romk(-/-) mice, confirming our recent observation that maxi-K channels contribute to distal K(+) secretion in the absence of ROMK. PMID- 18385267 TI - New insights into the function of the Wilms tumor suppressor gene WT1 in podocytes. AB - The Wilms tumor suppressor gene WT1 is essential for early urogenital development: homozygous mutations in WT1 result in embryonic lethality due to a failure in the development of kidneys and gonads. In the adult kidney, WT1 expression is limited to the glomerular podocytes. Several human nephrotic diseases arise from mutations of the WT1 gene, including mutations that affect its zinc-fingers and alternative splicing of +/- KTS isoforms. These include WAGR (for Wilms tumor, aniridia, genitourinary anomalies, and mental retardation), and Frasier and Denys-Drash syndromes. Recent advances including the development of transgenic mouse models and conditionally immortalized podocyte cell lines are beginning to shed light on WT1's crucial role in podocyte function. PMID- 18385268 TI - Epithelial Na+ channel activation and processing in mice lacking SGK1. AB - Amiloride-sensitive Na(+) channel activity was examined in the cortical collecting ducts of a mouse line (SGK1(-/-)) deficient in the serum- and glucocorticoid-dependent protein kinase SGK1. This activity was correlated with changes in renal Na handling and in the maturation of epithelial Na(+) channel (ENaC) protein. Neither SGK1(-/-) mice nor paired SGK1(+/+) animals expressed detectable channel activity, measured as amiloride-sensitive whole-cell current (I(Na)), under control conditions with standard chow. Administration of aldosterone (0.5 microg/h via osmotic minipump for 7 days) increased I(Na) to a similar extent in SGK1(+/+) (378 +/- 61 pA/cell at -100 mV) and in SGK1(-/-) (350 +/- 57 pA/cell) animals. However, the maturation of ENaC, assessed as the ratio of cleaved to full-length forms of gamma-ENaC, was more pronounced in SGK(+/+) mice. The SGK1(-/-) animals exhibited a salt-wasting phenotype when kept on a low Na diet for up to 2 days, losing significantly more Na in the urine than wild type mice. Under these conditions, I(Na) was enhanced more in SGK1(-/-) (94 +/- 14 pA/cell) than in SGK(+/+) (23 +/- 5 pA/cell) genotypes. Despite the larger currents, the ratio of cleaved to full-length gamma-ENaC was lower in the knockout animals. The mice also expressed a smaller amount of Na(+)-Cl(-) cotransporter protein under Na-depleted conditions. These results indicated that SGK1 is essential for optimal processing of ENaC but is not required for activation of the channel by aldosterone. PMID- 18385270 TI - Morphine induces mesangial cell proliferation and glomerulopathy via kappa-opioid receptors. AB - Morphine sulfate (MS) stimulates mesangial cell (MC) proliferation, a process central to development of glomerular disease. The purpose of this study was to examine whether specific opioid receptors (OR) and signal transducer and activators of transcription 3 (STAT3) signaling are associated with MS-induced MC proliferation. C57Bl/6J and OR-specific knockout (KO) mice were treated for up to 6 wk with PBS, MS (0.7-2.14 mg/kg), naloxone (equimolar to MS), or MS+naloxone (n = 6 per group). Glomerular volume and expression of PCNA, Thy1, and ED1/CD68 were analyzed in kidney sections. Cell proliferation and STAT3 phosphorylation were analyzed by bromodeoxyuridine (BrdU) ELISA and Western blot, respectively, in MCs in vitro. MS treatment led to enlarged kidneys and glomerulopathy and naloxone reversed these effects. MS treatment increased glomerular volume in both mu-OR (MOR) KO and delta-OR (DOR) KO mice, but not in kappa-OR (KOR) KO mice. To ascertain that MS-induced glomerulopathy in vivo was due to MC proliferation, we further examined the OR-specific effects of MS in MCs in vitro. MS-induced MC proliferation in vitro was inhibited by KOR-specific nor-BNI, but not by DOR or MOR-specific antagonists naltrindol or CTOP, respectively. KOR-specific agonist U50488H stimulated proliferation of MCs, but DOR-specific agonist DPDPE and MOR specific agonist DAMGO did not. MS failed to stimulate proliferation of MCs from KOR KO mice. MS and KOR agonists induced STAT3 phosphorylation, and STAT3 inhibitor blocked KOR agonist-induced MC proliferation. We show that MS stimulates glomerulopathy and MC proliferation via KOR and STAT3 signaling. PMID- 18385269 TI - Signaling pathways modulated by fish oil in salt-sensitive hypertension. AB - Although many studies have indicated that fish oil (FO) improves cardiovascular risk factors and reduces histopathological manifestations of injury in experimental renal injury models, potential mechanisms underlying this protective effect have not been adequately defined. The objective of this study was to identify potential signaling pathways that confer protection in the Dahl rat model of salt-sensitive hypertension. Male Dahl salt-sensitive rats (n = 10/group) were provided with formulated diets containing 8% NaCl, 20% protein, and 25% FO or 25% corn oil (CO) for 28 days. FO reduced blood pressure (-11% at 4 wk; P < 0.05), urine protein excretion (-45% at 4 wk; P < 0.05), plasma cholesterol and triglyceride levels (-54%, P < 0.001; and -58%, P < 0.05), and histopathological manifestations of renal injury, including vascular hypertrophy, segmental and global glomerular sclerosis, interstitial fibrosis, and tubular atrophy. Interstitial inflammation was significantly reduced by FO (-32%; P < 0.001), as assessed by quantitative analysis of ED1-positive cells in sections of the renal cortex. FO reduced tubulointerstitial proliferative activity, as assessed by Western blot analysis of cortical homogenates for PCNA (-51%; P < 0.01) and quantitative analysis of Mib-1-stained sections of the renal cortex ( 42%; P < 0.001). Decreased proliferative activity was associated with reduced phospho-ERK expression (-37%; P < 0.005) and NF-kappaB activation (-42%; P < 0.05). FO reduced cyclooxygenase (COX)-2 expression (-63%; P < 0.01) and membrane translocation of the NADPH oxidase subunits p47(phox) and p67(phox) (-26 and 34%; P < 0.05). We propose that FO ameliorates renal injury in Dahl salt sensitive rats through the inhibition of ERK, decreased NF-kappaB activation, inhibition of COX-2 expression, and decreased NADPH oxidase activation. PMID- 18385271 TI - Antiapoptotic properties of erythropoiesis-stimulating proteins in models of cisplatin-induced acute kidney injury. AB - Erythropoietin (Epo) induces erythrocytosis by suppressing erythroid progenitor cell apoptosis through the Janus-activated kinase-signal transducers and activators of transcription (JAK-STAT) pathway. Since apoptosis contributes to cisplatin (CP)-induced nephrotoxicity and Epo receptors (EpoR) are expressed in the kidney, we examined the role of antiapoptosis in recombinant human erythropoietin (rHuEpo)-mediated renal protection. In human renal proximal tubular epithelial (RPTE) cells in culture, rHuEpo, but not inactive rHuEpo (I rHuEpo), the receptor-binding sites of which are mutated, caused a significant reduction in CP-induced apoptosis at > or = 100 U/ml. rHuEpo, but not I-rHuEpo, increased STAT5 and Akt/PKB phosphorylation, demonstrating functional EpoR expression on RPTE cells. Furthermore, the JAK2 inhibitor tyrphostin AG-490 attenuated rHuEpo protection, suggesting a role of the JAK-STAT pathway in rHuEpo mediated antiapoptosis. In rats, intravenous administration of 5,000 U/kg rHuEpo, but not an equivalent peptide mass of I-rHuEpo, before a single 5.5 mg/kg iv injection of CP, significantly increased hematocrit (Hct) and reduced the CP induced increase in serum creatinine. Serum creatinine on day 4 was 3.4 +/- 0.3, 1.9 +/- 0.3, and 3.5 +/- 0.4 mg/dl in the CP, CP + rHuEpo, and CP + I-rHuEpo groups, respectively. Similarly, darbepoietin-alpha (DA), a hyperglycosylated analog of rHuEpo with prolonged in vivo activity when injected at 25 microg/kg iv before CP, significantly increased Hct and reduced serum creatinine. Renal clearance studies based on glomerular filtration rate and renal blood flow confirmed the significant renal protection by DA against CP. Tubular apoptosis and necrosis were significantly reduced in the kidneys of the CP + DA vs. the CP + saline group. Moreover, the equalization of Hct by venesection did not abrogate the DA-mediated renal protection. Administration of DA 48 h after CP injection also conferred significant renal protection. Thus our experiments confirm a role for erythropoiesis-stimulating proteins, including the new analog DA, in limiting CP-induced nephrotoxicity and suggest that antiapoptosis via the Epo-EpoR interaction is an important mechanism for renal protection. PMID- 18385272 TI - An androgen-inducible proximal tubule-specific Cre recombinase transgenic model. AB - To facilitate the study of renal proximal tubules, we generated a transgenic mouse strain expressing an improved Cre recombinase (iCre) under the control of the kidney androgen-regulated protein (KAP) promoter. The transgene was expressed in the kidney of male mice but not in female mice. Treatment of female transgenic mice with androgen induced robust expression of the transgene in the kidney. We confirmed the presence of Cre recombinase activity and the cell specificity by breeding the KAP2-iCRE mice with ROSA26 reporter mice. X-Gal staining of kidney sections from male double transgenic mice showed robust staining in the epithelial cells of renal proximal tubules. beta-Gal staining in female mice became evident in proximal tubules after administration of androgen. This model of inducible Cre recombinase in the renal proximal tubule should provide a novel useful tool for studying the physiological significance of genes expressed in the renal proximal tubule. This has advantages over other current models where Cre recombinase expression is constitutive, not inducible. PMID- 18385273 TI - Increased inorganic phosphate induces human endothelial cell apoptosis in vitro. AB - Chronic kidney disease with hyperphosphatemia is associated with accelerated atherosclerosis and endothelial dysfunction. However, the contribution of high serum phosphate levels to endothelial injury is incompletely understood. The aim of this work was to evaluate the responses of endothelial cells to elevated levels of extracellular phosphate in vitro. High phosphate in concentrations similar to those observed in uremia-associated hyperphosphatemia (>2.5 mM) induced apoptosis in two endothelial cell lines (EAhy926 cells and GM-7373 cells). This effect was enhanced when cells were incubated for 24 h in the presence of 2.8 mM calcium instead of 1.8 mM. By treating cells with 0.5 or 1.0 mM phosphonoformic acid, an inhibitor of the phosphate transporter, death was completely prevented. The process of phosphate-induced apoptosis was further characterized by increased oxidative stress, as detected by increased ROS generation and disruption of the mitochondrial membrane potential at approximately 2 h after treatment, followed by caspase activation. These findings show that hyperphosphatemia causes endothelial cell apoptosis, a process that impairs endothelial integrity. Endothelial cell injury induced by high phosphate concentrations may be an initial event leading to vascular complications in patients with chronic kidney disease. PMID- 18385274 TI - New binary polymorphisms reshape and increase resolution of the human Y chromosomal haplogroup tree. AB - Markers on the non-recombining portion of the human Y chromosome continue to have applications in many fields including evolutionary biology, forensics, medical genetics, and genealogical reconstruction. In 2002, the Y Chromosome Consortium published a single parsimony tree showing the relationships among 153 haplogroups based on 243 binary markers and devised a standardized nomenclature system to name lineages nested within this tree. Here we present an extensively revised Y chromosome tree containing 311 distinct haplogroups, including two new major haplogroups (S and T), and incorporating approximately 600 binary markers. We describe major changes in the topology of the parsimony tree and provide names for new and rearranged lineages within the tree following the rules presented by the Y Chromosome Consortium in 2002. Several changes in the tree topology have important implications for studies of human ancestry. We also present demography independent age estimates for 11 of the major clades in the new Y chromosome tree. PMID- 18385275 TI - Nonrecurrent MECP2 duplications mediated by genomic architecture-driven DNA breaks and break-induced replication repair. AB - Recurrent submicroscopic genomic copy number changes are the result of nonallelic homologous recombination (NAHR). Nonrecurrent aberrations, however, can result from different nonexclusive recombination-repair mechanisms. We previously described small microduplications at Xq28 containing MECP2 in four male patients with a severe neurological phenotype. Here, we report on the fine-mapping and breakpoint analysis of 16 unique microduplications. The size of the overlapping copy number changes varies between 0.3 and 2.3 Mb, and FISH analysis on three patients demonstrated a tandem orientation. Although eight of the 32 breakpoint regions coincide with low-copy repeats, none of the duplications are the result of NAHR. Bioinformatics analysis of the breakpoint regions demonstrated a 2.5 fold higher frequency of Alu interspersed repeats as compared with control regions, as well as a very high GC content (53%). Unexpectedly, we obtained the junction in only one patient by long-range PCR, which revealed nonhomologous end joining as the mechanism. Breakpoint analysis in two other patients by inverse PCR and subsequent array comparative genomic hybridization analysis demonstrated the presence of a second duplicated region more telomeric at Xq28, of which one copy was inserted in between the duplicated MECP2 regions. These data suggest a two-step mechanism in which part of Xq28 is first inserted near the MECP2 locus, followed by breakage-induced replication with strand invasion of the normal sister chromatid. Our results indicate that the mechanism by which copy number changes occur in regions with a complex genomic architecture can yield complex rearrangements. PMID- 18385276 TI - The development of the L-QoL: a quality-of-life instrument specific to systemic lupus erythematosus. AB - OBJECTIVES: Complex diseases, such as systemic lupus erythematosus (SLE), present dilemmas over choice of outcome measures. Using a battery of instruments to capture the impact of different impairments or activity limitations experienced does not provide assessment of the wider impact on quality of life (QoL). This paper describes the development and testing of a new instrument to measure QoL in systemic lupus erythematosus (L-QoL). METHODS: The development combines theoretical strengths of the needs-based QoL model with statistical and diagnostic powers of the Rasch model. Content was derived from in-depth interviews with relevant patients. Cognitive debriefing interviews assessed face and content validity. Rasch analysis was applied to data from an initial postal survey to remove misfitting items. A second postal survey assessed scaling properties, reliability, internal consistency and validity. RESULTS: A 55-item questionnaire was derived from interview transcripts. Cognitive debriefing confirmed acceptability. Rasch analysis of postal survey data (n = 95) removed misfitting items. A second postal survey (n = 93), produced a 25-item version with good item fit and stability, excellent test-retest reliability (0.92), internal consistency (0.92) and strict unidimensionality. CONCLUSIONS: It is concluded that the L-QoL should prove a valuable instrument for assessing patient based outcome in clinical trials and practice. PMID- 18385277 TI - Rates of new-onset psoriasis in patients with rheumatoid arthritis receiving anti tumour necrosis factor alpha therapy: results from the British Society for Rheumatology Biologics Register. AB - BACKGROUND: Anti-tumour necrosis factor (TNF)alpha treatments improve outcome in severe rheumatoid arthritis (RA) and are efficacious in psoriasis and psoriatic arthritis. However recent case reports describe psoriasis occurring as an adverse event in patients with RA receiving anti-TNFalpha therapy. OBJECTIVES: We aimed to determine whether the incidence rate of psoriasis was higher in patients with RA treated with anti-TNFalpha therapy compared to those treated with traditional disease-modifying antirheumatic drugs (DMARDs). We also compared the incidence rates of psoriasis between the three anti-TNFalpha drugs licensed for RA. METHODS: We studied 9826 anti-TNF-treated and 2880 DMARD-treated patients with severe RA from The British Society for Rheumatology Biologics Register (BSRBR). All patients reported with new onset psoriasis as an adverse event were included in the analysis. Incidence rates of psoriasis were calculated as events/1000 person years and compared using incidence rate ratios (IRR). RESULTS: In all, 25 incident cases of psoriasis in patients receiving anti-TNFalpha therapy and none in the comparison cohort were reported between January 2001 and July 2007. The absence of any cases in the comparison cohort precluded a direct comparison; however the crude incidence rate of psoriasis in those treated with anti-TNFalpha therapy was elevated at 1.04 (95% CI 0.67 to 1.54) per 1000 person years compared to the rate of 0 (upper 97.5% CI 0.71) per 1000 person years in the patients treated with DMARDs. Patients treated with adalimumab had a significantly higher rate of incident psoriasis compared to patients treated with etanercept (IRR 4.6, 95% CI 1.7 to 12.1) and infliximab (IRR 3.5, 95% CI 1.3 to 9.3). CONCLUSIONS: Results from this study suggest that the incidence of psoriasis is increased in patients treated with anti-TNFalpha therapy. Our findings also suggest that the incidence may be higher in patients treated with adalimumab. PMID- 18385278 TI - Aspiration of normal or asymptomatic pathological joints for diagnosis and research: indications, technique and success rate. AB - Although joint aspiration is a basic clinical skill, aspiration of normal joints, or asymptomatic clinically quiescent joints, is only rarely undertaken. There are two main indications for this procedure. Firstly, for definitive diagnosis of crystal-associated arthritis (gout and pseudogout) during the intercritical period and for subsequent monitoring of treatment success of gout; and secondly, to obtain normal synovial fluid for biomarker research. The justification for these indications, the success rate and the technical aspects related to this procedure are presented in this article. PMID- 18385279 TI - The tumour necrosis factor receptor superfamily member 1b 676T>G polymorphism in relation to response to infliximab and adalimumab treatment and disease severity in rheumatoid arthritis. AB - OBJECTIVE: To assess the effect of a functional polymorphism (676T>G, M196R) in the tumour necrosis factor receptor super family 1b (TNFSF1b) gene on disease activity, radiological joint damage and response to infliximab and adalimumab treatment in patients with rheumatoid arthritis (RA). METHODS: Two cohorts of patients with RA were genotyped for the 676T>G polymorphism (rs1061622) in exon 6 of the TNFSF1b gene by restriction fragment length polymorphism analysis. One cohort (n = 234) included patients from the Dutch Rheumatoid Arthritis Monitoring register with detailed information on their response to anti-TNF therapy (infliximab and adalimumab), the other cohort comprised patients from a long-term observational early inception cohort at our centre (n = 248). RESULTS: The 676T>G polymorphism was not associated with anti-TNF response after 3 or 6 months of treatment. Linear regression analysis showed no significant difference in the progression of radiological joint damage during the first 3 and 6 years of disease between the three genotype groups (TT, TG and GG). Additionally, no difference in mean disease activity between genotypes was seen after 3 and 6 years of disease. CONCLUSION: Despite its demonstrated functionality, the 676T>G polymorphism in the TNFSF1b gene does not have a major role in either the response to anti-TNF therapy or in the disease severity or radiological progression in RA. PMID- 18385280 TI - Surface change of the mammalian lens during accommodation. AB - Classical theories suggest that the surface area of the crystalline lens changes during accommodation while the lens volume remains constant. Our recent work challenged this view by showing that the lens volume decreases as the lens flattens during unaccommodation. In this paper we investigate 1) the magnitude of changes in the surface of the in vitro isolated cow lens during simulated accommodation, as well as that of human lens models, determined from lateral photographs and the application of the first theorem of Pappus; and 2) the velocity of the equatorial diameter recovery of prestretched cow and rabbit lenses by using a custom-built software-controlled stretching apparatus synchronized to a digital camera. Our results showed that the in vitro cow lens surface changed in an unexpected manner during accommodation depending on how much tension was applied to flatten the lens. In this case, the anterior surface initially collapsed with a reduction in surface followed by an increase in surface, when the stretching was applied. In the human lens model, the surface increased when the lens unaccommodated. The lens volume always decreases as the lens flattens. An explanation for the unexpected surface change is presented and discussed. Furthermore, we determined that the changes in lens volume, as reflected by the speed of the equatorial diameter recovery in in vitro cow and rabbit lenses during simulated accommodation, occurred within a physiologically relevant time frame (200 ms), implying a rapid movement of fluid to and from the lens during accommodation. PMID- 18385281 TI - NO-induced regulation of human trabecular meshwork cell volume and aqueous humor outflow facility involve the BKCa ion channel. AB - Nitric oxide (NO) donors decrease intraocular pressure (IOP) by increasing aqueous outflow facility in the trabecular meshwork (TM) and/or Schlemm's canal. However, the cellular mechanisms are unknown. Cellular mechanisms known to regulate outflow facility include changes in cell volume and cellular contractility. In this study, we investigated the effects of NO donors on outflow facility and NO-induced effects on TM cell volume. We tested the involvement of soluble guanylate cyclase (sGC), cGMP, PKG, and the large-conductance Ca2+ activated K+ (BKCa) channel using inhibitors and activators. Cell volume was measured using calcein AM fluorescent dye, detected by confocal microscopy, and quantified using NIH ImageJ software. An anterior segment organ perfusion system measured outflow facility. NO increased outflow facility in porcine eye anterior segments (0.4884-1.3956 microl.min(-1).mmHg(-1)) over baseline (0.2373-0.5220 microl.min(-1).mmHg(-1)) within 10 min of drug application. These NO-induced increases in outflow facility were inhibited by the the BKCa channel inhibitor IBTX. Exposure of TM cells to NO resulted in a 10% decrease in cell volume, and these decreases were abolished by the sGC inhibitor 1H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one and IBTX, suggesting the involvement of sGC and K+ eflux, respectively. NO-induced decreases in cell volume were mimicked by 8-Br-cGMP and abolished by the PKG inhibitor (RP)-8-Br-PET-cGMP-S, suggesting the involvement cGMP and PKG. Additionally, the time course for NO-induced decreases in TM cell volume correlated with NO-induced increases in outflow facility, suggesting that the NO-induced alterations in cell volume may influence outflow facility. PMID- 18385282 TI - CaM kinase II delta2-dependent regulation of vascular smooth muscle cell polarization and migration. AB - Previous studies indicate involvement of the multifunctional Ca2+/calmodulin dependent protein kinase II (CaMKII) in vascular smooth muscle (VSM) cell migration. In the present study, molecular loss-of-function studies were used specifically to assess the role of the predominant CaMKII delta2 isoform on VSM cell migration using a scratch wound healing assay. Targeted CaMKII delta2 knockdown using siRNA or inhibition of activity by overexpressing a kinase negative mutant resulted in attenuation of VSM cell migration. Temporal and spatial assessments of kinase autophosphorylation indicated rapid and transient activation in response to wounding, in addition to a sustained activation in the leading edge of migrating and spreading cells. Furthermore, siRNA-mediated suppression of CaMKII delta2 resulted in the inhibition of wound-induced Rac activation and Golgi reorganization, and disruption of leading edge morphology, indicating an important function for CaMKII delta2 in regulating VSM cell polarization. Numerous previous reports link activation of CaMKII to ERK1/2 signaling in VSM. Wound-induced ERK1/2 activation was also found to be dependent on CaMKII; however, ERK activity did not account for effects of CaMKII in regulating Golgi polarization, indicating alternative mechanisms by which CaMKII affects the complex events involved in cell migration. Wounding a VSM cell monolayer results in CaMKII delta2 activation, which positively regulates VSM cell polarization and downstream signaling, including Rac and ERK1/2 activation, leading to cell migration. PMID- 18385283 TI - Apical adenosine regulates basolateral Ca2+-activated potassium channels in human airway Calu-3 epithelial cells. AB - In airway epithelial cells, apical adenosine regulates transepithelial anion secretion by activation of apical cystic fibrosis transmembrane conductance regulator (CFTR) via adenosine receptors and cAMP/PKA signaling. However, the potent stimulation of anion secretion by adenosine is not correlated with its modest intracellular cAMP elevation, and these uncorrelated efficacies have led to the speculation that additional signaling pathways may be involved. Here, we showed that mucosal adenosine-induced anion secretion, measured by short-circuit current (Isc), was inhibited by the PLC-specific inhibitor U-73122 in the human airway submucosal cell line Calu-3. In addition, the Isc was suppressed by BAPTA AM (a Ca2+ chelator) and 2-aminoethoxydiphenyl borate (2-APB; an inositol 1,4,5 trisphosphate receptor blocker), but not by PKC inhibitors, suggesting the involvement of PKC-independent PLC/Ca2+ signaling. Ussing chamber and patch-clamp studies indicated that the adenosine-induced PLC/Ca2+ signaling stimulated basolateral Ca2+-activated potassium (KCa) channels predominantly via A2B adenosine receptors and contributed substantially to the anion secretion. Thus, our data suggest that apical adenosine activates contralateral K+ channels via PLC/Ca2+ and thereby increases the driving force for transepithelial anion secretion, synergizing with its modulation of ipsilateral CFTR via cAMP/PKA. Furthermore, the dual activation of CFTR and KCa channels by apical adenosine resulted in a mixed secretion of chloride and bicarbonate, which may alter the anion composition in the secretion induced by secretagogues that elicit extracellular ATP/adenosine release. Our findings provide novel mechanistic insights into the regulation of anion section by adenosine, a key player in the airway surface liquid homeostasis and mucociliary clearance. PMID- 18385284 TI - Neuroinflammation facilitates LIF entry into brain: role of TNF. AB - Leukemia inhibitory factor (LIF) is a proinflammatory cytokine mediating a variety of central nervous system (CNS) responses to inflammatory stimuli. During lipopolysaccharide (LPS)-induced inflammation, blood concentrations of LIF increase, correlating with lethality of sepsis. Circulating LIF crosses the blood brain barrier (BBB) by a saturable transport system. Here we determine how this transport system is regulated in neuroinflammation. Using transport assays that quantify the influx rate and volume of distribution of LIF in mice, we show that LPS facilitated the permeation of LIF from the blood to the brain without compromising the paracellular permeability of the BBB as determined by coadministration of fluorescein. Concurrently, gp130 (shared by the interleukin-6 family of cytokines), but not gp190 (the specific receptor for LIF) or cilliary neutrophic factor (CNTF-Ralpha, a unique receptor for cilliary neurotrophic factor that also uses gp130 and gp190), showed increased levels of mRNA and protein expression in cerebral microvessels from the LPS-treated mice. The upregulation of gp130 by LPS was at least partially mediated by vascular tumor necrosis factor receptor (TNFR)1 and TNFR2. This was shown by elevated TNFR1 and TNFR2 mRNA and protein in cerebral microvessels after LPS and by the absence of the LPS effect on gp130 in knockout mice lacking these receptors. The results show that neuroinflammation by LPS induces endothelial signaling and enhances cytokine transport across the BBB. PMID- 18385285 TI - A Kv channel with an altered activation gate sequence displays both "fast" and "slow" activation kinetics. AB - The Kv1-4 families of K+ channels contain a tandem proline motif (PXP) in the S6 helix that is crucial for channel gating. In human Kv1.5, replacing the first proline by an alanine resulted in a nonfunctional channel. This mutant was rescued by introducing another proline at a nearby position, changing the sequence into AVPP. This resulted in a channel that activated quickly (ms range) upon the first depolarization. However, thereafter, the channel became trapped in another gating mode that was characterized by slow activation kinetics (s range) with a shallow voltage dependence. The switch in gating mode was observed even with very short depolarization steps, but recovery to the initial "fast" mode was extremely slow. Computational modeling suggested that switching occurred during channel deactivation. To test the effect of the altered PXP sequence on the mobility of the S6 helix, we used molecular dynamics simulations of the isolated S6 domain of wild type (WT) and mutants starting from either a closed or open conformation. The WT S6 helix displayed movements around the PXP region with simulations starting from either state. However, the S6 with a AVPP sequence displayed flexibility only when started from the closed conformation and was rigid when started from the open state. These results indicate that the region around the PXP motif may serve as a "hinge" and that changing the sequence to AVPP results in channels that deactivate to a state with an alternate configuration that renders them "reluctant" to open subsequently. PMID- 18385286 TI - Aberrant cell-to-cell coupling in Ca2+-overloaded guinea pig ventricular muscles. AB - To investigate how intercellular coupling can be changed during Ca2+ overloading of ventricular muscle, we studied Ca2+ signals in individual cells and the histochemistry of the major gap junction channel, connexin43 (Cx43), using multicellular preparations. Papillary muscles were obtained from guinea pig ventricles and loaded with rhod-2. Sequential Ca2+ images of surface cells were obtained with a confocal microscope. In intact muscles, all cells showed simultaneous Ca2+ transients in response to field stimulation over a field of view of 0.3 x 0.3 mm2. In severely Ca2+-overloaded muscles, obtained by high frequency stimulation in nonflowing Krebs solution, cells became less responsive to stimulation. Furthermore, nonsimultaneous but serial onsets of Ca2+ transients were often detected, suggesting a propagation delay of action potentials. The time lag of the onset between two aligned cells was sometimes as long as 100 ms. Similar lags were also observed in muscles with gap junction channels inhibited by heptanol. To investigate whether the phosphorylation state of Cx43 is affected in Ca2+-overloaded muscles, the distributions of phosphorylated and nonphosphorylated Cx43 were determined using specific antibodies. Most of the Cx43 was phosphorylated in the nonoverloaded muscles, whereas nonphosphorylated Cx43 was significantly elevated in severely Ca2+-overloaded muscles. Our results suggest that the propagation delay of action potential within a small area, a few square millimeters, can be a cause of abnormal conduction and a microreentry in Ca2+-overloaded heart. Inactivation of Na+ channels and inhibition of gap junctional communication may underlie the cell-to-cell propagation delay. PMID- 18385287 TI - Asymmetric dimethylarginine inhibits HSP90 activity in pulmonary arterial endothelial cells: role of mitochondrial dysfunction. AB - Increased asymmetric dimethylarginine (ADMA) levels have been implicated in the pathogenesis of a number of conditions affecting the cardiovascular system. However, the mechanism(s) by which ADMA exerts its effect has not been adequately elucidated. Thus the purpose of this study was to determine the effect of increased ADMA on nitric oxide (NO) signaling and to begin to elucidate the mechanism by which ADMA acts. Our initial data demonstrated that ADMA increased NO synthase (NOS) uncoupling in both recombinant human endothelial NO synthase (eNOS) and pulmonary arterial endothelial cells (PAEC). Furthermore, we found that this endothelial NOS (eNOS) uncoupling increased 3-nitrotyrosine levels preferentially in the mitochondria of PAEC due to a redistribution of eNOS from the plasma membrane to the mitochondria. This increase in nitration in the mitochondria was found to induce mitochondrial dysfunction as determined by increased mitochondrial-derived reactive oxygen species and decreased generation of ATP. Finally, we found that the decrease in ATP resulted in a reduction in the chaperone activity of HSP90 resulting in a decrease in its interaction with eNOS. In conclusion increased levels of ADMA causes mitochondrial dysfunction and a loss of heat shock protein-90 chaperone activity secondary to an uncoupling of eNOS. Mitochondrial dysfunction may be an understudied component of the endothelial dysfunction associated with various cardiovascular disease states. PMID- 18385289 TI - Activation of cytosolic phospholipase A2 and fatty acid transacylase is essential but not sufficient for thrombin-induced smooth muscle cell proliferation. AB - Thrombin is a potent stimulant of smooth muscle cell (SMC) proliferation in inflammatory conditions, leading to pathological thickening of vascular walls in atherosclerosis and airway remodeling in asthma. Cell proliferation requires the formation and remodeling of cell membrane phospholipids (PLs), involving the activation of PL-metabolizing enzymes. Yet, the role of specific PL-metabolizing enzymes in SMC proliferation has hardly been studied. To bridge this gap, in the present study, we investigated the role of key enzymes involved in PL metabolism, the PL-hydrolyzing enzyme phospholipase A2 (PLA2) and the PL-synthesizing enzyme lysophosphatidic acid-fatty acid transacylase (LPAAT), in thrombin-induced proliferation of bovine aortic SMCs (BASMCs). Concomitantly with the induction of BASMC proliferation, thrombin activated cytosolic PLA2 (cPLA2-alpha), expressed by selective release of arachidonic acid and mRNA expression, as well as LPAAT, expressed by nonselective incorporation of fatty acid and mRNA expression. Specific inhibitors of these enzymes, arachidonyl-trifluoromethyl-ketone for cPLA2 and thimerosal for LPAAT, suppressed their activities, concomitantly with suppression of BASMC proliferation, suggesting a mandatory requirement for cPLA2 and LPAAT activation in thrombin-induced SMC proliferation. Thrombin acts through the protease-activated receptor (PAR-1), and, accordingly, we found that thrombin induced BASMC proliferation was suppressed by the PAR-1 inhibitor SCH-79797. However, the PAR-1 inhibitor did not prevent thrombin-induced mRNA expression of cPLA2 and LPAAT, implying that the activation of cPLA2 and LPAAT is essential but not sufficient for thrombin-induced proliferation of BASMCs. PMID- 18385288 TI - Endogenous heparan sulfate and heparin modulate bone morphogenetic protein-4 signaling and activity. AB - Bone morphogenetic proteins (BMPs) and their endogenous antagonists are important for brain and bone development and tumor initiation and progression. Heparan sulfate (HS) proteoglycans (HSPG) modulate the activities of BMPs and their antagonists. How glycosaminoglycans (GAGs) influence BMP activity in various malignancies and in inherited abnormalities of GAG metabolism, and the structural features of GAGs essential for modulation of BMP signaling, remain incompletely defined. We examined whether chemically modified soluble heparins, the endogenous HS in malignant cells and the HS accumulated in Hurler syndrome cells influence BMP-4 signaling and activity. We show that both exogenous (soluble) and endogenous GAGs modulate BMP-4 signaling and activity, and that this effect is dependent on specific sulfate residues of GAGs. Our studies suggest that endogenous sulfated GAGs promote the proliferation and impair differentiation of malignant human cells, providing the rationale for investigating whether pharmacological agents that inhibit GAG synthesis or function might reverse this effect. Our demonstration of impairment of BMP-4 signaling by GAGs in multipotent stem cells in human Hurler syndrome identifies a mechanism that might contribute to the progressive neurological and skeletal abnormalities in Hurler syndrome and related mucopolysaccharidoses. PMID- 18385290 TI - Distinct pathways of ERK activation by the muscarinic agonists pilocarpine and carbachol in a human salivary cell line. AB - Cholinergic-muscarinic receptor agonists are used to alleviate mouth dryness, although the cellular signals mediating the actions of these agents on salivary glands have not been identified. We examined the activation of ERK1/2 by two muscarinic agonists, pilocarpine and carbachol, in a human salivary cell line (HSY). Immunoblot analysis revealed that both agonists induced transient activation of ERK1/2. Whereas pilocarpine induced phosphorylation of the epidermal growth factor (EGF) receptor, carbachol did not. Moreover, ERK activation by pilocarpine, but not carbachol, was abolished by the EGF receptor inhibitor AG-1478. Downregulation of PKC by prolonged treatment of cells with the phorbol ester PMA diminished carbachol-induced ERK phosphorylation but had no effect on pilocarpine responsiveness. Depletion of intracellular Ca2+ ([Ca2+]i by EGTA did not affect ERK activation by either agent. In contrast to carbachol, pilocarpine did not elicit [Ca2+]i mobilization in HSY cells. Treatment of cells with the muscarinic receptor subtype 3 (M3) antagonist N-(3-chloropropyl)-4 piperidnyl diphenylacetate decreased ERK responsiveness to both agents, whereas the subtype 1 (M1) antagonist pirenzepine reduced only the carbachol response. Stimulation of ERKs by pilocarpine was also decreased by M3, but not M1, receptor small interfering RNA. The Src inhibitor PP2 blocked pilocarpine-induced ERK activation and EGF receptor phosphorylation, without affecting ERK activation by carbachol. Our results demonstrate that the actions of pilocarpine and carbachol in salivary cells are mediated through two distinct signaling mechanisms pilocarpine acting via M3 receptors and Src-dependent transactivation of EGF receptors, and carbachol via M1/M3 receptors and PKC-converging on the ERK pathway. PMID- 18385291 TI - Depletion of beta-COP reveals a role for COP-I in compartmentalization of secretory compartments and in biosynthetic transport of caveolin-1. AB - We have utilized small interfering RNA (siRNA)-mediated depletion of the beta-COP subunit of COP-I to explore COP-I function in organellar compartmentalization and protein traffic. Reduction in beta-COP levels causes the colocalization of markers for the endoplasmic reticulum (ER)-Golgi intermediate compartment (ERGIC), Golgi, trans-Golgi network (TGN), and recycling endosomes in large, globular compartments. The lack of spatial differentiation of these compartments is not due to a general collapse of all cellular organelles since markers for the early endosomes and lysosomes do not redistribute to the common structures. Anterograde trafficking of the transmembrane cargo vesicular stomatitis virus membrane glycoprotein and of a subset of soluble cargoes is arrested within the common globular compartments. Similarly, recycling traffic of transferrin through the common compartment is perturbed. Furthermore, the trafficking of caveolin-1 (Cav1), a structural protein of caveolae, is arrested within the globular structures. Importantly, Cav1 coprecipitates with the gamma-subunit of COP-I, suggesting that Cav1 is a COP-I cargo. Our findings suggest that COP-I is required for the compartmentalization of the ERGIC, Golgi, TGN, and recycling endosomes and that COP-I plays a novel role in the biosynthetic transport of Cav1. PMID- 18385292 TI - Evaluation of HepaRG cells as an in vitro model for human drug metabolism studies. AB - HepaRG cells, a newly developed human hepatoma cell line, differentiate into hepatocyte-like morphology by treatment with dimethyl sulfoxide (DMSO). The expression of cytochrome P450 (P450) enzymes, transporter proteins, and transcription factors was stable in differentiated HepaRG cells over a period of 6 weeks when cultured with DMSO. Compared with human hepatocytes, expression of P450 in HepaRG cells was in general lower with the exception for a considerably higher expression of CYP3A4 and CYP7A1. The expression of P450s generally decreased when DMSO was removed from the medium, whereas transporters and liver specific factors were unaffected. The relative mRNA content of drug-metabolizing P450s displayed the highest resemblance between human hepatocytes and differentiated HepaRG cells 1 day after removal of DMSO from the medium. The metabolism of midazolam, naloxone, and clozapine in HepaRG cells was similar to human hepatocytes, indicating the function of CYP3A4, CYP1A2, and UDP glucuronosyltransferase enzymes. However, the metabolism of 7-ethoxycoumarin and dextromethorphan was low, confirming low levels of CYP2E1 and CYP2D6 in HepaRG cells. The P450 probe substrates indicate a decrease in CYP1A2, CYP2B6, CYP2C9, and CYP3A4 activities in HepaRG cells 1 day after removal of DMSO from the medium. The activities were then relatively stable in DMSO-free medium for up to 14 days. Based on the stable expression of liver-specific functions over a long period in culture, the relative mRNA content of drug-metabolizing P450s, and metabolic properties, HepaRG cells provide a valuable in vitro model for human drug metabolism studies. PMID- 18385294 TI - Counterpoint: High-frequency ventilation is not the optimal physiological approach to ventilate ARDS patients. PMID- 18385293 TI - Effects of spice constituents on P-glycoprotein-mediated transport and CYP3A4 mediated metabolism in vitro. AB - The effects of eight components from six commonly consumed spices on P glycoprotein (P-gp) transport and CYP3A4 metabolism were evaluated in vitro. P-gp mediated [(3)H]digoxin fluxes across the L-MDR1 (LLC-PK1 cells transfected with human MDR1 gene) and Caco-2 (human colon carcinoma) cell monolayers showed a marked asymmetry compared with that in the LLC-PK1 (porcine kidney epithelial cells) cell monolayers. Curcumin (from turmeric) at 30 to 60 microM and 6 gingerol (from ginger) at 100 to 500 microM were observed to inhibit P-gp mediated [(3)H]digoxin transport in L-MDR1 and Caco-2 cells. Effects of spices on midazolam (MDZ) 1'-hydroxylation and 4-hydroxylation of CYP3A4 activity were determined in pooled human liver microsomes (HLM). The following IC(50) values for effects of spices on MDZ 1'-hydroxylation in HLM were obtained: 29 microM for curcumin, 1.17 mM for allyl methyl disulfide (AMD) (from Chinese chive), 1.02 mM for 1,8-cineole (from coriander), and 1.28 mM for beta-caryophyllene (from curry leaf). CYP3A4-mediated 4-hydroxylation of MDZ was inhibited by curcumin at 30, 45, and 60 microM (4-hydroxy-MDZ formation was decreased to 52, 30, and 29%, respectively, compared with control), by 6-gingerol at 60, 100, and 500 microM (71, 68, and 38%), by AMD at 1 and 4 mM (29 and 14%), by d-limonene (from coriander) at 4 mM (65%), by 1,8-cineole at 0.5, 1, and 4 mM (74, 64, and 59%), and by citral (from lemongrass) at 1 mM (59%). Among the spices that showed inhibitory effect on MDZ metabolism in HLM, only AMD showed a preincubation time dependent inhibitory effect on MDZ metabolism in HLM, suggesting the AMD as an irreversible CYP3A4 inhibitor. PMID- 18385297 TI - High-frequency ventilation is/is not the optimal physiological approach to ventilate ARDS patients. PMID- 18385298 TI - Last word on point:counterpoint: High-frequency ventilation is/is not the optimal physiological approach to ventilate ARDS patients. PMID- 18385299 TI - Last word on point:counterpoint: High-frequency ventilation is/is not the optimal physiological approach to ventilate ARDS patients. PMID- 18385300 TI - Sickle cell trait should/should not be considered asymptomatic and as a benign condition during physical activity. PMID- 18385301 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385302 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385303 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385304 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385305 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385306 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385307 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385308 TI - Commentary on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385309 TI - Last word on viewpoint: Perspective on the future use of genomics in exercise prescription. PMID- 18385310 TI - Bradykinin: a microglia attractant in vivo? PMID- 18385311 TI - Thalamic shape: a possible endophenotype. PMID- 18385312 TI - Glycinergic and GABA(A)-mediated inhibition of somatic motoneurons does not mediate rapid eye movement sleep motor atonia. AB - A hallmark of rapid eye movement (REM) sleep is a potent suppression of postural muscle tone. Motor control in REM sleep is unique because it is characterized by flurries of intermittent muscle twitches that punctuate muscle atonia. Because somatic motoneurons are bombarded by strychnine-sensitive IPSPs during REM sleep, it is assumed that glycinergic inhibition underlies REM atonia. However, it has never been determined whether glycinergic inhibition of motoneurons is indeed responsible for triggering the loss of postural muscle tone during REM sleep. Therefore, we used reverse microdialysis, electrophysiology, and pharmacological and histological methods to determine whether glycinergic and/or GABA(A)-mediated neurotransmission at the trigeminal motor pool mediates masseter muscle atonia during REM sleep in rats. By antagonizing glycine and GABA(A) receptors on trigeminal motoneurons, we unmasked a tonic glycinergic/GABAergic drive at the trigeminal motor pool during waking and non-rapid eye movement (NREM) sleep. Blockade of this drive potently increased masseter muscle tone during both waking and NREM sleep. This glycinergic/GABAergic drive was immediately switched-off and converted into a phasic glycinergic drive during REM sleep. Blockade of this phasic drive potently provoked muscle twitch activity in REM sleep; however, it did not prevent or reverse REM atonia. Muscle atonia in REM even persisted when glycine and GABA(A) receptors were simultaneously antagonized and trigeminal motoneurons were directly activated by glutamatergic excitation, indicating that a powerful, yet unidentified, inhibitory mechanism overrides motoneuron excitation during REM sleep. Our data refute the prevailing hypothesis that REM atonia is caused by glycinergic inhibition. The inhibitory mechanism mediating REM atonia therefore requires reevaluation. PMID- 18385313 TI - Lasting syndrome of depression produced by reduction in serotonin uptake during postnatal development: evidence from sleep, stress, and behavior. AB - Dysfunction of the serotonin system is implicated in sleep and emotional disorders. To test whether these impairments could arise during development, we studied the impact of early-life, transient versus genetic, permanent alterations of serotonin reuptake on sleep-wakefulness patterns, depression-related behavior, and associated physiological features. Here, we show that female mice treated neonatally with a highly selective serotonin reuptake inhibitor, escitalopram, exhibited signs of depression in the form of sleep anomalies, anhedonia, increased helplessness reversed by chronic antidepressant treatment, enhanced response to acute stress, and increased serotoninergic autoinhibitory feedback. This syndrome was not reproduced by treatment in naive adults but resembled the phenotype of mutant mice lacking the serotonin transporter, except that these exhibited decreased serotonin autoreceptor sensitivity and additional anxiety like behavior. Thus, alteration of serotonin reuptake during development, whether induced by external or genetic factors, causes a depressive syndrome lasting into adulthood. Such early-life impairments might predispose individuals to sleep and/or mood disorders. PMID- 18385314 TI - A two-year study with fibrillar beta-amyloid (Abeta) immunization in aged canines: effects on cognitive function and brain Abeta. AB - Aged canines (dogs) accumulate human-type beta-amyloid (Abeta) in diffuse plaques in the brain with parallel declines in cognitive function. We hypothesized that reducing Abeta in a therapeutic treatment study of aged dogs with preexisting Abeta pathology and cognitive deficits would lead to cognitive improvements. To test this hypothesis, we immunized aged beagles (8.4-12.4 years) with fibrillar Abeta(1-42) formulated with aluminum salt (Alum) for 2.4 years (25 vaccinations). Cognitive testing during this time revealed no improvement in measures of learning, spatial attention, or spatial memory. After extended treatment (22 vaccinations), we observed maintenance of prefrontal-dependent reversal learning ability. In the brain, levels of soluble and insoluble Abeta(1-40) and Abeta(1 42) and the extent of diffuse plaque accumulation was significantly decreased in several cortical regions, with preferential reductions in the prefrontal cortex, which is associated with a maintenance of cognition. However, the amount of soluble oligomers remained unchanged. The extent of prefrontal Abeta was correlated with frontal function and serum anti-Abeta antibody titers. Thus, reducing total Abeta may be of limited therapeutic benefit to recovery of cognitive decline in a higher mammalian model of human brain aging and disease. Immunizing animals before extensive Abeta deposition and cognitive decline to prevent oligomeric or fibrillar Abeta formation may have a greater impact on cognition and also more directly evaluate the role of Abeta on cognition in canines. Alternatively, clearing preexisting Abeta from the brain in a treatment study may be more efficacious for cognition if combined with a second intervention that restores neuron health. PMID- 18385315 TI - Somatostatin receptor subtype 4 couples to the M-current to regulate seizures. AB - The K(+) M-current (I(M), Kv7) is an important regulator of cortical excitability, and mutations in these channels cause a seizure disorder in humans. The neuropeptide somatostatin (SST), which has antiepileptic properties, augments I(M) in hippocampal CA1 pyramidal neurons. We used SST receptor knock-out mice and subtype-selective ligands to investigate the receptor subtype that couples to I(M) and mediates the antiepileptic effects of SST. Using pentylenetetrazole as a chemoconvulsant, SST(2), SST(3), and SST(4) receptor knock-out mice all had shorter latencies to different seizure stages and increased seizure severity when compared with wild-type mice. However, the most robust differences were observed in the SST(4) knock-outs. When seizures were induced by systemic injection of kainate, only SST(4) knock-outs showed an increase in seizure sensitivity. We next examined the action of SST and subtype-selective SST agonists on electrophysiological parameters in hippocampal slices of wild-type and receptor knock-out mice. SST(2) and SST(4) appear to mediate the majority of SST inhibition of epileptiform activity in CA1. SST lacked presynaptic effects in mouse CA1, in contrast to our previous findings in rat. SST increased I(M) in CA1 pyramidal neurons of wild-type and SST(2) knock-out mice, but not SST(4) knock out mice. Using M-channel blockers, we found that SST(4) coupling to M-channels is critical to its inhibition of epileptiform activity. This is the first demonstration of an endogenous enhancer of I(M) that is important in controlling seizure activity. SST(4) receptors could therefore be an important novel target for developing new antiepileptic and antiepileptogenic drugs. PMID- 18385316 TI - In DRG11 knock-out mice, trigeminal cell death is extensive and does not account for failed brainstem patterning. AB - A previous study (Ding et al., 2003) showed that the homeodomain transcription factor DRG11 is necessary for pattern formation in the trigeminal nucleus principalis (PrV), the requisite brainstem nucleus for development of the whisker to-barrel cortex pathway. However, it is not known how DRG11 contributes to pattern formation. Anatomical studies were performed in DRG11 knock-out (-/-) and DRG11/Bax double -/- mice to test the hypotheses that DRG11 is required for neuronal survival in the V pathway and that PrV cell death is sufficient to explain pattern alterations. At birth, DRG11(-/-) mice had equivalent cell loss in the V ganglion, PrV, and spinal V subnucleus interpolaris (SpVi). Because whisker-related patterns were normal in the SpVi, cell death would not appear to explain failed pattern formation in the mutant PrV. Electron microscopy revealed exuberant apoptosis and necrosis as the mechanisms of PrV cell death occurring in the late prenatal and newborn DRG11(-/-), when such cell death was up to six times more prevalent than normal. DRG11 heterozygote and Bax(-/-) mice were crossed in an attempt to dissociate PrV patterning anomalies from exuberant apoptosis in DRG11(-/-) mice. Both DRG11(-/-) and DRG11/Bax double -/- mutants lacked whisker-related patterning in their PrV, despite Bax(-/-)-induced rescue of V ganglion and PrV cells. Thus, apoptotic cell death is not a sufficient cause of failed pattern formation in the PrV of the DRG11(-/-). A signaling pathway involving DRG11 may, therefore, be the elusive PrV pattern maker. PMID- 18385317 TI - Neurodevelopmental trajectories of the human cerebral cortex. AB - Understanding the organization of the cerebral cortex remains a central focus of neuroscience. Cortical maps have relied almost exclusively on the examination of postmortem tissue to construct structural, architectonic maps. These maps have invariably distinguished between areas with fewer discernable layers, which have a less complex overall pattern of lamination and lack an internal granular layer, and those with more complex laminar architecture. The former includes several agranular limbic areas, and the latter includes the homotypical and granular areas of association and sensory cortex. Here, we relate these traditional maps to developmental data from noninvasive neuroimaging. Changes in cortical thickness were determined in vivo from 764 neuroanatomic magnetic resonance images acquired longitudinally from 375 typically developing children and young adults. We find differing levels of complexity of cortical growth across the cerebrum, which align closely with established architectonic maps. Cortical regions with simple laminar architecture, including most limbic areas, predominantly show simpler growth trajectories. These areas have clearly identified homologues in all mammalian brains and thus likely evolved in early mammals. In contrast, polysensory and high-order association areas of cortex, the most complex areas in terms of their laminar architecture, also have the most complex developmental trajectories. Some of these areas are unique to, or dramatically expanded in primates, lending an evolutionary significance to the findings. Furthermore, by mapping a key characteristic of these development trajectories (the age of attaining peak cortical thickness) we document the dynamic, heterochronous maturation of the cerebral cortex through time lapse sequences ("movies"). PMID- 18385318 TI - RPM-1, a Caenorhabditis elegans protein that functions in presynaptic differentiation, negatively regulates axon outgrowth by controlling SAX-3/robo and UNC-5/UNC5 activity. AB - Changes in axon outgrowth patterns are often associated with synaptogenesis. Members of the conserved Pam/Highwire/RPM-1 protein family have essential functions in presynaptic differentiation. Here, we show that Caenorhabditis elegans RPM-1 negatively regulates axon outgrowth mediated by the guidance receptors SAX-3/robo and UNC-5/UNC5. Loss-of-function rpm-1 mutations cause a failure to terminate axon outgrowth, resulting in an overextension of the longitudinal PLM axon. We observe that PLM overextension in rpm-1 mutants is suppressed by sax-3 and unc-5 loss-of-function mutations. PLM axon overextension is also induced by SAX-3 overexpression, and the length of extension is enhanced by loss of rpm-1 function or suppressed by loss of unc-5 function. We also observe that loss of rpm-1 function in genetic backgrounds sensitized for guidance defects disrupts ventral AVM axon guidance in a SAX-3-dependent manner and enhances dorsal guidance of DA and DB motor axons in an UNC-5-dependent manner. Loss of rpm-1 function alters expression of the green fluorescent protein (GFP)-tagged proteins, SAX-3::GFP and UNC-5::GFP. RPM-1 is known to regulate axon termination through two parallel genetic pathways; one involves the Rab GEF (guanine nucleotide exchange factor) GLO-4, which regulates vesicular trafficking, and another that involves the F-box protein FSN-1, which mediates RPM-1 ubiquitin ligase activity. We show that glo-4 but not fsn-1 mutations affect axon guidance in a manner similar to loss of rpm-1 function. Together, the results suggest that RPM-1 regulates axon outgrowth affecting axon guidance and termination by controlling the trafficking of the UNC-5 and SAX-3 receptors to cell membranes. PMID- 18385319 TI - Protein 600 is a microtubule/endoplasmic reticulum-associated protein in CNS neurons. AB - There is an increasing body of literature pointing to cytoskeletal proteins as spatial organizers and interactors of organelles. In this study, we identified protein 600 (p600) as a novel microtubule-associated protein (MAP) developmentally regulated in neurons. p600 exhibits the unique feature to interact with the endoplasmic reticulum (ER). Silencing of p600 by RNA interference (RNAi) destabilizes neuronal processes in young primary neurons undergoing neurite extension and containing scarce staining of the ER marker Bip. Furthermore, in utero electroporation of p600 RNAi alters neuronal migration, a process that depends on synergistic actions of microtubule dynamics and ER functions. p600-depleted migrating neurons display thin, crooked, and "zigzag" leading process with very few ER membranes. Thus, p600 constitutes the only known MAP to associate with the ER in neurons, and this interaction may impact on multiple cellular processes ranging from neuronal development to neuronal maturation and plasticity. PMID- 18385320 TI - Interneuronal activity in reflex pathways from group II muscle afferents is monitored by dorsal spinocerebellar tract neurons in the cat. AB - The main aim of the study was to investigate whether group II muscle afferents contribute to the inhibition of dorsal spinocerebellar tract (DSCT) neurons and thereby modulate information provided by these neurons in the cat. In intracellular recordings, we found disynaptic IPSPs from group II afferents in the majority of DSCT neurons, most often in parallel with IPSPs evoked from group I afferents. In an attempt to identify interneurons that mediate these IPSPs, the second aim of the study, laminas IV-VII in midlumbar segments were searched for interneurons antidromically activated by stimuli applied within Clarke's column. Such interneurons were found in regions in which focal field potentials were evoked by group I and II afferents, or ventral to them, and most were coexcited by these afferents. The input to these interneurons and their location indicate that they belonged to the previously identified population of premotor interneurons in disynaptic pathways between group I and II afferents and hindlimb motoneurons. The study leads thus to the conclusion that inhibitory actions of group II afferents on DSCT neurons are collateral to actions on motoneurons and that DSCT neurons monitor inhibitory actions of group II afferents on motoneurons as closely as they monitor actions of group I afferents. The results also indicate that interneurons mediating disynaptic reflex actions from tendon organ (group Ib) afferents and those mediating disynaptic actions from secondary muscle spindle (group II) afferents to motoneurons may be parts and parcel of the same interneuronal population rather than constitute distinct interneuronal populations. PMID- 18385321 TI - NMDA receptor subunit NR2A is required for rapidly acquired spatial working memory but not incremental spatial reference memory. AB - NMDA receptors (NMDARs) containing NR2A (epsilon1) subunits are key contributors to hippocampal long-term potentiation (LTP) induction in adult animals and have therefore been widely implicated in hippocampus-dependent spatial learning. Here we show that mice lacking the NR2A subunit or its C-terminal intracellular domain exhibit impaired spatial working memory (SWM) but normal spatial reference memory (SRM). Both NR2A mutants acquired the SRM version of the water maze task, and the SRM component of the radial maze, as well as controls. They were, however, impaired on a non-matching-to-place T-maze task, and on the SWM component of the radial maze. In addition, NR2A knock-out mice displayed a diminished spatial novelty preference in a spontaneous exploration Y-maze task, and were impaired on a T-maze task in which distinctive inserts present on the floor of the maze determined which goal arm contained the reward, but only if there was a discontiguity between the conditional cue and the place at which the reward was delivered. This dissociation of spatial memory into distinctive components is strikingly similar to results obtained with mice lacking glutamate receptor-A (GluR-A)-containing AMPA receptors, which support long-term potentiation expression. These results identify a specific role for a NMDAR-dependent signaling pathway that leads to the activation of a GluR-A-dependent expression mechanism in a rapidly acquired, flexible form of spatial memory. This mechanism depends on the C-terminal intracellular domain of the NR2A subunit. In contrast, the ability to associate a particular spatial location with the water maze escape platform or food reward is NR2A independent, as well as GluR-A independent. PMID- 18385322 TI - Cyclin-dependent kinase 5 phosphorylation of human septin SEPT5 (hCDCrel-1) modulates exocytosis. AB - Cyclin-dependent kinase 5 (Cdk5) is predominantly expressed in the nervous system, where it is involved in neuronal migration, synaptic transmission, and survival. The role of Cdk5 in synaptic transmission is mediated by regulating the cellular functions of presynaptic proteins such as synapsin, Munc18, and dynamin 1. Its multifunctional role at the synapse is complex and probably involves other novel substrates. To explore this possibility, we used a yeast two-hybrid screen of a human cDNA library with p35 as bait and isolated human septin 5 (SEPT5), known also as hCDCrel-1, as an interacting clone. Here we report that p35 associates with SEPT5 in GST (glutathione S-transferase)-pull-down and coimmunoprecipitation assays. We confirmed that Cdk5/p35 phosphorylates SEPT5 in vitro and in vivo and identified S327 of SEPT5 as a major phosphorylation site. A serine (S)-to-alanine (A) 327 mutant of SEPT5 bound syntaxin more efficiently than SEPT5 wild type. Additionally, coimmunoprecipitation from synaptic vesicle fractions and Cdk5 wild-type and knock-out lysates showed that phosphorylation of septin 5 by Cdk5/p35 decreases its binding to syntaxin-1. Moreover, mutant nonphosphorylated SEPT5 potentiated regulated exocytosis more than the wild type when each was expressed in PC12 cells. These data suggest that Cdk5 phosphorylation of human septin SEPT5 at S327 plays a role in modulating exocytotic secretion. PMID- 18385323 TI - Effects on differentiation of embryonic ventral midbrain progenitors by Lmx1a, Msx1, Ngn2, and Pitx3. AB - Neurons derived from neural stem cells could potentially be used for cell therapy in neurodegenerative disorders, such as Parkinson's disease. To achieve controlled differentiation of neural stem cells, we expressed transcription factors involved in the development of midbrain dopaminergic neurons in rat and human neural progenitors. Using retroviral-mediated transgene delivery, we overexpressed Lmx1a (LIM homeobox transcription factor 1, alpha), Msx1 (msh homeobox homolog 1), Ngn2 (neurogenin 2), or Pitx3 (paired-like homeodomain transcription factor 3) in neurospheres derived from embryonic day 14.5 rat ventral mesencephalic progenitors. We also expressed either Lmx1a or Msx1 in the human embryonic midbrain-derived progenitor cell line NGC-407. Rat cells transduced with Ngn2 exited the cell cycle and expressed the neuronal marker microtubule-associated protein 2 and catecholamine-neuron protein vesicular monoamine transporter 2. Interestingly, Pitx3 downregulated the expression of SOX2 (SRY-box containing gene 2) and Nestin, altered cell morphology, but never induced neuronal or glial differentiation. Ngn2 exhibited a strong neuron inducing effect. In contrast, few Lmx1a-transduced cells matured into neurons, and Msx1 overexpression promoted oligodendrogenesis rather than neuronal differentiation. Importantly, none of these four genes, alone or in combination, enhanced differentiation of rat neural stem cells into dopaminergic neurons. Notably, the overexpression of Lmx1a, but not Msx1, in human neural progenitors increased the yield of tyrosine hydroxylase-immunoreactive cells by threefold. Together, we demonstrate that induced overexpression of transcription factor genes has profound and specific effects on the differentiation of rat and human midbrain progenitors, although few dopamine neurons are generated. PMID- 18385324 TI - Functional gradients of auditory sensitivity along the anterior ectosylvian sulcus of the cat. AB - Determining the spatial direction of sound sources is one of the major computations performed by the auditory system. The anterior ectosylvian sulcus (AES) of cat cortex is known to be important for sound localization. However, there are contradicting reports as to the spatial response properties of neurons in AES: whereas some studies found narrowly tuned neurons, others reported mostly spatially widely tuned neurons. We hypothesized that this is the result of a nonhomogenous distribution of the auditory neurons in this area. To test this possibility, we recorded neuronal activity along the AES, together with a sample of neurons from primary auditory cortex (A1) of cats in response to pure tones and to virtual acoustic space stimuli. In all areas, most neurons responded to both types of stimuli. Neurons located in posterior AES (pAES) showed special response properties that distinguished them from neurons in A1 and from neurons in anterior AES (aAES). The proportion of space-selective neurons among auditory neurons was significantly higher in pAES (82%) than in A1 (72%) and in aAES (60%). Furthermore, whereas the large majority of A1 neurons responded preferentially to contralateral sounds, neurons in pAES (and to a lesser extent in aAES) had their spatial selectivity distributed more homogenously. In particular, 28% of the space-selective neurons in pAES had highly modulated frontal receptive fields, against 8% in A1 and 17% in aAES. We conclude that in cats, pAES contains a secondary auditory cortical field which is specialized for spatial processing, in particular for the representation of frontal space. PMID- 18385326 TI - Adeno-associated virus transfer of a gene encoding SNAP-25 resistant to botulinum toxin A attenuates neuromuscular paralysis associated with botulism. AB - Advances in viral gene therapy have opened new possibilities for treating a range of motor neuron diseases, but these have not yet been translated into clinically applicable therapies because of difficulties in delivery to susceptible/damaged neurons, ambiguities in the identity of gene(s) implicated, and a paucity of means to quantify any physiological improvement. Most of these hurdles can be overcome by using the neuromuscular paralysis induced by botulinum neurotoxin type A (BoNT/A) as a prototype disease. Furthermore, because human botulism, occasionally fatal, causes prolonged muscle disablement as a result of the intraneuronal persistence of the toxin's SNAP-25 (S25)-cleaving protease, development of a genetic approach could lead to a potential treatment for this debilitating disease. Adeno-associated viral delivery of a cleavage-resistant S25 gene (S25-R198T) to chromaffin cells in vitro yielded exocytotically active S25 R198T that diminished subsequent blockade by BoNT/A of evoked catecholamine release. Evaluation in vivo, by administering this virus into rat spinal cord before injecting BoNT/A, showed a decreased inhibition of acetylcholine release as reflected in elevated retention of neuromuscular transmission. A similar, although smaller, protection of synaptic transmission from the toxin was seen after peripherally injecting the therapeutic virus. Such therapy also curtailed nerve sprouting normally induced by BoNT/A. This first demonstration of the utility of a DNA-based therapy for botulism paves the way for further advances in its treatment and for application to genetic disorders of motor neurons. PMID- 18385327 TI - Long-distance retrograde effects of botulinum neurotoxin A. AB - Botulinum neurotoxins (designated BoNT/A-BoNT/G) are bacterial enzymes that block neurotransmitter release by cleaving essential components of the vesicle fusion machinery. BoNT/A, which cleaves SNAP-25 (synaptosomal-associated protein of 25 kDa), is extensively exploited in clinical medicine to treat neuromuscular pathologies, facial wrinkles, and various types of pain. It is widely assumed that BoNT/A remains at the synaptic terminal and its effects are confined to the injection site. Here we demonstrate that catalytically active BoNT/A is retrogradely transported by central neurons and motoneurons and is then transcytosed to afferent synapses, in which it cleaves SNAP-25. SNAP-25 cleavage by BoNT/A was observed in the contralateral hemisphere after unilateral BoNT/A delivery to the hippocampus. Appearance of cleaved SNAP-25 resulted in blockade of hippocampal activity in the untreated hemisphere. Injections of BoNT/A into the optic tectum led to the appearance of BoNT/A-truncated SNAP-25 in synaptic terminals within the retina. Cleaved SNAP-25 also appeared in the facial nucleus after injection of the toxin into rat whisker muscles. Experiments excluded passive spread of the toxin and demonstrated axonal migration and neuronal transcytosis of BoNT/A. These findings reveal a novel pathway of BoNT/A trafficking in neurons and have important implications for the clinical uses of this neurotoxin. PMID- 18385325 TI - Presynaptic calcium channel localization and calcium-dependent synaptic vesicle exocytosis regulated by the Fuseless protein. AB - A systematic forward genetic Drosophila screen for electroretinogram mutants lacking synaptic transients identified the fuseless (fusl) gene, which encodes a predicted eight-pass transmembrane protein in the presynaptic membrane. Null fusl mutants display >75% reduction in evoked synaptic transmission but, conversely, an approximately threefold increase in the frequency and amplitude of spontaneous synaptic vesicle fusion events. These neurotransmission defects are rescued by a wild-type fusl transgene targeted only to the presynaptic cell, demonstrating a strictly presynaptic requirement for Fusl function. Defects in FM dye turnover at the synapse show a severely impaired exo-endo synaptic vesicle cycling pool. Consistently, ultrastructural analyses reveal accumulated vesicles arrested in clustered and docked pools at presynaptic active zones. In the absence of Fusl, calcium-dependent neurotransmitter release is dramatically compromised and there is little enhancement of synaptic efficacy with elevated external Ca(2+) concentrations. These defects are causally linked with severe loss of the Cacophony voltage-gated Ca(2+) channels, which fail to localize normally at presynaptic active zone domains in the absence of Fusl. These data indicate that Fusl regulates assembly of the presynaptic active zone Ca(2+) channel domains required for efficient coupling of the Ca(2+) influx and synaptic vesicle exocytosis during neurotransmission. PMID- 18385328 TI - Dopamine depletion impairs frontostriatal functional connectivity during a set shifting task. AB - We investigated the effect of transient dopamine depletion on functional connectivity during performance of the Wisconsin Card Sorting Task. Functional magnetic resonance imaging data were analyzed as a psychophysiological interaction, a statistical method used to identify functional connectivity during experimental manipulations. Nineteen healthy subjects were scanned, double blind, on 2 separate days: once after drinking an amino acid mixture deficient in the dopamine precursors tyrosine and phenylalanine, and once after drinking a nutritionally balanced mixture. In the balanced drink session, statistically significant connectivity between the frontal lobes and striatum was observed during set shifting, and the greater the prefrontostriatal connectivity, the faster the response time after a shift. Neither of these associations were observed after dopamine depletion. Moreover, dopamine depletion also reduced the degree of deactivation in areas normally suppressed during attention-demanding tasks, including the medial prefrontal cortex, posterior cingulate cortex, and hippocampus. Together, these results suggest that functional connectivity between the frontal lobes and basal ganglia during set shifting contributes to more efficient performance and that dopamine modulates this corticostriatal connectivity. PMID- 18385329 TI - Intermediate progenitors in adult hippocampal neurogenesis: Tbr2 expression and coordinate regulation of neuronal output. AB - Neurogenesis in the adult hippocampus is a highly regulated process that originates from multipotent progenitors in the subgranular zone (SGZ). Currently, little is known about molecular mechanisms that regulate proliferation and differentiation in the SGZ. To study the role of transcription factors (TFs), we focused on Tbr2 (T-box brain gene 2), which has been implicated previously in developmental glutamatergic neurogenesis. In adult mouse hippocampus, Tbr2 protein and Tbr2-GFP (green fluorescent protein) transgene expression were specifically localized to intermediate-stage progenitor cells (IPCs), a type of transit amplifying cells. The Tbr2+ IPCs were highly responsive to neurogenic stimuli, more than doubling after voluntary wheel running. Notably, the Tbr2+ IPCs formed cellular clusters, the average size of which (Tbr2+ cells per cluster) likewise more than doubled in runners. Conversely, Tbr2+ IPCs were selectively depleted by antimitotic drugs, known to suppress neurogenesis. After cessation of antimitotic treatment, recovery of neurogenesis was paralleled by recovery of Tbr2+ IPCs, including a transient rebound above baseline numbers. Finally, Tbr2 was examined in the context of additional TFs that, together, define a TF cascade in embryonic neocortical neurogenesis (Pax6 --> Ngn2 --> Tbr2 --> NeuroD --> Tbr1). Remarkably, the same TF cascade was found to be linked to stages of neuronal lineage progression in adult SGZ. These results suggest that Tbr2+ IPCs play a major role in the regulation of adult hippocampal neurogenesis, and that a similar transcriptional program controls neurogenesis in adult SGZ as in embryonic cerebral cortex. PMID- 18385331 TI - Brain chromatin remodeling: a novel mechanism of alcoholism. AB - The treatment of alcoholism requires the proper management of ethanol withdrawal symptoms, such as anxiety, to prevent further alcohol use and abuse. In this study, we investigated the potential role of brain chromatin remodeling, caused by histone modifications, in alcoholism. We found that the anxiolytic effects produced by acute alcohol were associated with a decrease in histone deacetylase (HDAC) activity and increases in acetylation of histones (H3 and H4), levels of CREB (cAMP-responsive element binding) binding protein (CBP), and neuropeptide Y (NPY) expression in the amygdaloid brain regions of rats. However, the anxiety like behaviors during withdrawal after chronic alcohol exposure were associated with an increase in HDAC activity and decreases in acetylation of H3 and H4, and levels of both CBP and NPY in the amygdala. Blocking the observed increase in HDAC activity during alcohol withdrawal with the HDAC inhibitor, trichostatin A, rescued the deficits in H3 and H4 acetylation and NPY expression (mRNA and protein levels) in the amygdala (central and medial nucleus of amygdala) and prevented the development of alcohol withdrawal-related anxiety in rats as measured by the elevated plus maze and light/dark box exploration tests. These results reveal a novel role for amygdaloid chromatin remodeling in the process of alcohol addiction and further suggest that HDAC inhibitors may be potential therapeutic agents in treating alcohol withdrawal symptoms. PMID- 18385330 TI - Working memory for social cues recruits orbitofrontal cortex and amygdala: a functional magnetic resonance imaging study of delayed matching to sample for emotional expressions. AB - During everyday interactions, we continuously monitor and maintain information about different individuals and their changing emotions in memory. Yet to date, working memory (WM) studies have primarily focused on mechanisms for maintaining face identity, but not emotional expression, and studies investigating the neural basis of emotion have focused on transient activity, not delay related activity. The goal of this functional magnetic resonance imaging study was to investigate WM for two critical social cues: identity and emotion. Subjects performed a delayed match-to-sample task that required them to match either the emotional expression or the identity of a face after a 10 s delay. Neuroanatomically, our predictions focused on the orbitofrontal cortex (OFC) and the amygdala, as these regions have previously been implicated in emotional processing and long-term memory, and studies have demonstrated sustained OFC and medial temporal lobe activity during visual WM. Consistent with previous studies, transient activity during the sample period representing emotion and identity was found in the superior temporal sulcus and inferior occipital cortex, respectively. Sustained delay-period activity was evident in OFC, amygdala, and hippocampus, for both emotion and identity trials. These results suggest that, although initial processing of emotion and identity is accomplished in anatomically segregated temporal and occipital regions, sustained delay related memory for these two critical features is held by the OFC, amygdala and hippocampus. These regions share rich connections, and have been shown previously to be necessary for binding features together in long-term memory. Our results suggest a role for these regions in active maintenance as well. PMID- 18385332 TI - Protein kinase A-induced phosphorylation of the p65 subunit of nuclear factor kappaB promotes Schwann cell differentiation into a myelinating phenotype. AB - Axon-Schwann cell interactions are critical for myelin formation during peripheral nerve development and regeneration. Axonal contact promotes Schwann cell precursors to differentiate into a myelinating phenotype, and cAMP-elevating agents can mimic this; however, the mechanisms underlying this differentiation are poorly understood. We demonstrated previously that the transcription factor nuclear factor-kappaB (NF-kappaB) is required for myelin formation by Schwann cells (Nickols et al., 2003), although how it is activated during this process remained to be determined. Here, we report that culturing Schwann cells with sensory neurons results in the activation of cAMP-dependent protein kinase (PKA), and this kinase phosphorylates the p65 subunit of NF-kappaB at S276. The phosphorylation was also induced in cultured Schwann cells by treatment with forskolin, dibutyryl-cAMP, or by overexpression of a catalytic subunit of PKA, and this increased the transcriptional activity of NF-kappaB. In developing perinatal rat sciatic nerve, the kinetics of p65 phosphorylation at S276 paralleled that of PKA and NF-kappaB activation. To elucidate the role of p65 phosphorylation in myelin formation, we overexpressed an S276A mutant of p65 in cultured Schwann cells, which blocked PKA-mediated transcriptional activation of NF-kappaB. When the Schwann cells expressing the mutant were cocultured with sensory neurons, there was a 45% reduction in the number of myelinated fibers relative to controls, demonstrating a requirement for p65 phosphorylation by PKA during myelin formation. PMID- 18385333 TI - A rate code for sound azimuth in monkey auditory cortex: implications for human neuroimaging studies. AB - Is sound location represented in the auditory cortex of humans and monkeys? Human neuroimaging experiments have had only mixed success at demonstrating sound location sensitivity in primary auditory cortex. This is in apparent conflict with studies in monkeys and other animals, in which single-unit recording studies have found stronger evidence for spatial sensitivity. Does this apparent discrepancy reflect a difference between humans and animals, or does it reflect differences in the sensitivity of the methods used for assessing the representation of sound location? The sensitivity of imaging methods such as functional magnetic resonance imaging depends on the following two key aspects of the underlying neuronal population: (1) what kind of spatial sensitivity individual neurons exhibit and (2) whether neurons with similar response preferences are clustered within the brain. To address this question, we conducted a single-unit recording study in monkeys. We investigated the nature of spatial sensitivity in individual auditory cortical neurons to determine whether they have receptive fields (place code) or monotonic (rate code) sensitivity to sound azimuth. Second, we tested how strongly the population of neurons favors contralateral locations. We report here that the majority of neurons show predominantly monotonic azimuthal sensitivity, forming a rate code for sound azimuth, but that at the population level the degree of contralaterality is modest. This suggests that the weakness of the evidence for spatial sensitivity in human neuroimaging studies of auditory cortex may be attributable to limited lateralization at the population level, despite what may be considerable spatial sensitivity in individual neurons. PMID- 18385334 TI - Deconstruction of spatial integrity in visual stimulus detected by modulation of synchronized activity in cat visual cortex. AB - Spatiotemporal relationships among contour segments can influence synchronization of neural responses in the primary visual cortex. We performed a systematic study to dissociate the impact of spatial and temporal factors in the signaling of contour integration via synchrony. In addition, we characterized the temporal evolution of this process to clarify potential underlying mechanisms. With a 10 x 10 microelectrode array, we recorded the simultaneous activity of multiple cells in the cat primary visual cortex while stimulating with drifting sine-wave gratings. We preserved temporal integrity and systematically degraded spatial integrity of the sine-wave gratings by adding spatial noise. Neural synchronization was analyzed in the time and frequency domains by conducting cross-correlation and coherence analyses. The general association between neural spike trains depends strongly on spatial integrity, with coherence in the gamma band (35-70 Hz) showing greater sensitivity to the change of spatial structure than other frequency bands. Analysis of the temporal dynamics of synchronization in both time and frequency domains suggests that spike timing synchronization is triggered nearly instantaneously by coherent structure in the stimuli, whereas frequency-specific oscillatory components develop more slowly, presumably through network interactions. Our results suggest that, whereas temporal integrity is required for the generation of synchrony, spatial integrity is critical in triggering subsequent gamma band synchronization. PMID- 18385335 TI - Glutamatergic contributions to nicotinic acetylcholine receptor agonist-evoked cholinergic transients in the prefrontal cortex. AB - Because modulation of cortical cholinergic neurotransmission has been hypothesized to represent a necessary mechanism mediating the beneficial cognitive effects of nicotine and nicotinic acetylcholine receptor (nAChR) subtype-selective agonists, we used choline-sensitive microelectrodes for the real-time measurement of ACh release in vivo, to characterize cholinergic transients evoked by nicotine and the alpha4beta2*-selective nAChR partial agonist 2-methyl-3-(2-(S)-pyrrolindinylmethoxy)pyridine dihydrochloride (ABT 089), a clinically effective cognition enhancer. In terms of cholinergic signal amplitudes, ABT-089 was significantly more potent than nicotine in evoking ACh cholinergic transients. Moreover, cholinergic signals evoked by ABT-089 were characterized by faster signal rise time and decay rate. The nAChR antagonist mecamylamine attenuated the cholinergic signals evoked by either compound. Cholinergic signals evoked by ABT-089 were more efficaciously attenuated by the relatively beta2*-selective nAChR antagonist dihydro-beta-erythroidine. The alpha7 antagonist methyllycaconitine did not affect choline signal amplitudes but partly attenuated the relatively slow decay rate of nicotine-evoked cholinergic signals. Furthermore, the AMPA receptor antagonist DNQX as well as the NMDA receptor antagonist APV more potently attenuated cholinergic signals evoked by ABT-089. Using glutamate-sensitive microelectrodes to measure glutamatergic transients, ABT-089 was more potent than nicotine in evoking glutamate release. Glutamatergic signals were highly sensitive to tetrodotoxin-induced blockade of voltage-regulated sodium channels. Together, the present evidence indicates that compared with nicotine, ABT-089 evokes more potent and sharper cholinergic transients in prefrontal cortex. Glutamatergic mechanisms necessarily mediate the cholinergic effects of nAChR agonists in the prefrontal cortex. PMID- 18385336 TI - Synaptic defects in a Drosophila model of congenital muscular dystrophy. AB - The congenital muscular dystrophies present in infancy with muscle weakness and are often associated with mental retardation. Many of these inherited disorders share a common etiology: defective O-glycosylation of alpha-dystroglycan, a component of the dystrophin complex. Protein-O-mannosyl transferase 1 (POMT1) is the first enzyme required for the glycosylation of alpha-dystroglycan, and mutations in the POMT1 gene can lead to both Walker-Warburg syndrome (WWS) and limb girdle muscular dystrophy type 2K (LGMD2K). WWS is associated with severe mental retardation and major structural abnormalities in the brain; however, LGMD2K patients display a more mild retardation with no obvious structural defects in the brain. In a screen for synaptic mutants in Drosophila, we identified mutations in the Drosophila ortholog of POMT1, dPOMT1. Because synaptic defects are a plausible cause of mental retardation, we investigated the molecular and physiological defects associated with loss of dPOMT1 in Drosophila. In dPOMT1 mutants, there is a decrease in the efficacy of synaptic transmission and a change in the subunit composition of the postsynaptic glutamate receptors at the neuromuscular junction. We demonstrate that dPOMT1 is required to glycosylate the Drosophila dystroglycan ortholog Dg in vivo, and that this is the likely cause of these synaptic defects because (1) mutations in Dg lead to similar synaptic defects and (2) genetic interaction studies suggest that dPOMT1 and Dg function in the same pathway. These results are consistent with the model that dPOMT1-dependent glycosylation of Dg is necessary for proper synaptic function and raise the possibility that similar synaptic defects occur in the congenital muscular dystrophies. PMID- 18385337 TI - Artificial synaptic conductances reduce subthreshold oscillations and periodic firing in stellate cells of the entorhinal cortex. AB - Previous work has established that stellate cells of the medial entorhinal cortex produce prominent intrinsic subthreshold oscillations in the voltage response concentrated within the theta range (3-7 Hz). It has been speculated that these oscillations play an important role in vivo in establishing network behavior both in the entorhinal cortex and hippocampus. Consequently, it is important to investigate under what conditions theta oscillations in stellate cells can be generated and whether the spike-train power spectral density (PSD) also carries power at theta. We investigated the ability of stellate cells to generate theta oscillations in the presence of generic in vivo-like patterns of stimulation. Inputs were Poisson process-driven excitatory and inhibitory synaptic conductances or currents, introduced via dynamic clamp. We analyzed the subthreshold membrane oscillations and spike-train behavior in the presence of comparable synaptic conductance- or current-mediated membrane fluctuations. In the presence of conductance-based synapses, subthreshold oscillations are highly attenuated or entirely eliminated. Conversely, with current-based synapses stellate cells retain their ability to generate subthreshold oscillations in the theta band. These results also extend into the spiking regime, where only under current-based synapses does the PSD of the spike train show a prominent peak at theta. Furthermore, the peak in the spike-train PSD and spike clustering results from an increased probability of firing after a spike afterhyperpolarization and not directly from subthreshold oscillatory dynamics as has been previously suggested. Our results suggest that subthreshold oscillations may contribute less to in vivo response properties than has been hypothesized. PMID- 18385338 TI - Subventricular zone-mediated ependyma repair in the adult mammalian brain. AB - The subventricular zone (SVZ) of the adult mouse brain is a narrow stem cell niche that lies along the length of the lateral wall of the lateral ventricles. The SVZ supports neurogenesis throughout adulthood; however, with increasing age, the ventral SVZ deteriorates and only the dorsolateral SVZ remains neurogenic. Associated with the elderly dorsolateral SVZ, we reported previously an increased number of astrocytes interposed within the adjacent ependymal lining. Here, we show that astrocytes integrated within the ependyma are dividing, BrdU-labeled astrocytes that share cellular adherens with neighboring ependymal cells. By tracking BrdU-labeled astrocytes over time, we observed that, as they incorporated within the ependyma, they took on antigenic and morphologic characteristics of ependymal cells, suggesting a novel form of SVZ-supported "regenerative" repair in the aging brain. A similar form of SVZ-mediated ependyma repair was also observed in young mice after mild ependymal cell denudation with low dosages of neuraminidase. Together, this work identifies a novel non-neuronal mechanism of regenerative repair by the adult SVZ. PMID- 18385339 TI - Self-assembling nanofibers inhibit glial scar formation and promote axon elongation after spinal cord injury. AB - Peptide amphiphile (PA) molecules that self-assemble in vivo into supramolecular nanofibers were used as a therapy in a mouse model of spinal cord injury (SCI). Because self-assembly of these molecules is triggered by the ionic strength of the in vivo environment, nanoscale structures can be created within the extracellular spaces of the spinal cord by simply injecting a liquid. The molecules are designed to form cylindrical nanofibers that display to cells in the spinal cord the laminin epitope IKVAV at nearly van der Waals density. IKVAV PA nanofibers are known to inhibit glial differentiation of cultured neural stem cells and to promote neurite outgrowth from cultured neurons. In this work, in vivo treatment with the PA after SCI reduced astrogliosis, reduced cell death, and increased the number of oligodendroglia at the site of injury. Furthermore, the nanofibers promoted regeneration of both descending motor fibers and ascending sensory fibers through the lesion site. Treatment with the PA also resulted in significant behavioral improvement. These observations demonstrate that it is possible to inhibit glial scar formation and to facilitate regeneration after SCI using bioactive three-dimensional nanostructures displaying high densities of neuroactive epitopes on their surfaces. PMID- 18385340 TI - Dorsal-ventral gradient for neuronal plasticity in the embryonic spinal cord. AB - Within the developing Xenopus spinal cord, voltage-gated potassium (Kv) channel genes display different expression patterns, many of which occur in opposing dorsal-ventral gradients. Regional differences in Kv gene expression would predict different patterns of potassium current (I(Kv)) regulation. However, during the first 24 h of postmitotic differentiation, all primary spinal neurons undergo a temporally coordinated upregulation of I(Kv) density that shortens the duration of the action potential. Here, we tested whether spinal neurons demonstrate regional differences in I(Kv) regulation subsequent to action potential maturation. We show that two types of neurons, I and II, can be identified in culture on the basis of biophysical and pharmacological properties of I(Kv) and different firing patterns. Chronic increases in extracellular potassium, a signature of high neuronal activity, do not alter excitability properties of either neuron type. However, elevating extracellular potassium acutely after the period of action potential maturation leads to different changes in membrane properties of the two types of neurons. I(Kv) of type I neurons gains sensitivity to the blocker XE991, whereas type II neurons increase I(Kv) density and fire fewer action potentials. Moreover, by recording from neurons in vivo, we found that primary spinal neurons can be identified as either type I or type II. Type I neurons predominate in dorsal regions, whereas type II neurons localize to ventral regions. The findings reveal a dorsal-ventral gradient for I(Kv) regulation and a novel form of neuronal plasticity in spinal cord neurons. PMID- 18385341 TI - Missing the forest for the trees in biomedical research: the example of noma. PMID- 18385342 TI - Malaria vector management: where have we come from and where are we headed? PMID- 18385343 TI - Schistosomiasis in a 43-year-old African immigrant from Mali with invasive squamous cell carcinoma of the urinary bladder. PMID- 18385344 TI - Noma in Laos: stigma of severe poverty in rural Asia. AB - Noma, or cancrum oris, is a debilitating necrotizing ulcerative stomatitis that destroys the mouth and face. It usually starts in early childhood and is associated with severe poverty, malnutrition, and infections. It is most frequently described from sub-Saharan Africa but is under-reported. There have been very few reports from Asia. We describe the clinical and social features of a series of 12 patients with noma from remote poor villages in rural Lao People's Democratic Republic (Laos). Noma is an ominous stigma of severe poverty and the description of this disease emphasizes the importance of poverty reduction and nutritional improvement in Lao development. In the meantime, more awareness of the problem and the importance of early therapy in acute noma by primary health care workers may reduce mortality and prevent progression to severe disfigurement. PMID- 18385345 TI - Dihydroartemisinin-piperaquine rescue treatment of multidrug-resistant Plasmodium falciparum malaria in pregnancy: a preliminary report. AB - Dihydroartemisinin-piperaquine (DHA-PPQ) is a promising new artemisinin combination treatment. There are no published data on the intentional use of the drug in pregnancy. Between June 2006 and January 2007, 50 Karen pregnant women with recurrent P. falciparum infections, despite 7-day treatments with quinine or artesunate (+/-clindamycin) or both, were treated with DHA-PPQ. This rescue treatment was effective and well tolerated and there was no evidence of toxicity for the mothers or the fetus. The PCR adjusted cure rate by Kaplan Meier analysis at day 63 was 92.2% (95% CI: 76.9-97.4). PMID- 18385346 TI - Encephalopathy after ivermectin treatment in a patient infected with Loa loa and Plasmodium spp. AB - Despite over 350 million people being safely treated with ivermectin, there have been rare cases of death post-treatment; these events are most often associated with high Loa loa microfilaremia. This first autopsy description of an encephalopathy case following the administration of ivermectin involves a 45-year old male who became comatose 3 days after treatment. He slowly deteriorated over 5 weeks and died at 54 days after the anthelminthic treatment, probably as a result of a secondary skin or pulmonary infection exacerbated by malnutrition. The major pre- and post-autopsy findings included the presence of high loads of Loa loa, positivity for Plasmodium, the presence of a longstanding respiratory condition, and vascular pathology in the brain. The central nervous system lesions have similarities with those described in previously reported cases of Loa loa-associated death following diethylcarbamazine treatment. PMID- 18385347 TI - Impaired clinical response in a patient with uncomplicated falciparum malaria who received poor-quality and underdosed intramuscular artemether. AB - We describe an adult with uncomplicated Plasmodium falciparum malaria who did not improve clinically despite 5 days of intramuscular artemether therapy. He was prescribed a lower dose (kg body weight) than that recommended, and a vial from the packet contained only 74% of the artemether dose as stated by the manufacturer. The combination of underdosing, poor-quality drug, and the intrinsic low bioavailability of artemether may have contributed to his poor clinical response. Analysis of the packaging and chemical "fingerprinting" of the artemether suggested that the drug was genuine but was either substandard or had deteriorated after manufacture. PMID- 18385348 TI - Pathophysiology of jaundice in amoebic liver abscess. AB - Jaundice in patients with amoebic liver abscess is a frequent occurrence. However, the pathophysiology of jaundice in these patients is not fully understood. Hepatic necrosis leads to damage to bile ducts as well as various vascular structures, which in turn leads to biliovascular fistula and jaundice. We studied the mechanism of jaundice in patients with amoebic liver abscess. We prospectively evaluated 12 patients with amoebic liver abscess and jaundice from February 2002 to August 2007. All patients underwent various investigations, including imaging studies. There were 11 males and 1 female patient with a mean age of 41.3 years. Mean duration of illness before presentation was 13.8 days. All patients had fever and jaundice. We detected damaged hepatic veins and bile ducts in all patients with amoebic liver abscess causing biliovascular fistula and hyperbilirubinemia, which reverted to normal after biliary diversion with nasobiliary drainage. Jaundice in patients with amoebic liver abscess is caused by biliovascular fistula resulting from hepatic necrosis leading to damage to bile ducts and hepatic veins. PMID- 18385349 TI - Seroconversion to filarial antigens in Australian defence force personnel in Timor-Leste. AB - To investigate whether Australian soldiers were exposed to filarial parasites that cause lymphatic filariasis during a 6-month deployment to Timor-Leste, antifilarial antibody levels were measured in 907 soldiers using an enzyme linked immunosorbent assay (ELISA). Initial testing using Dirofilaria immitis antigen demonstrated that 49 of 907 (5.4%) soldiers developed antifilarial antibodies of the IgG1 subclass after deployment, whereas 1 of 944 (0.1%) seroconverted to the IgG4 subclass. When a sub sample of 88 D. immitis-reactive sera was subject to testing with an antifilarial antibody test using Brugia malayi antigen, 46 had elevated IgG antibodies, whereas 5 had elevated antibodies of the IgG4 subclass. A total of 24 soldiers seroconverted to B. malayi, as measured by parasite specific IgG, whereas 1 seroconverted to IgG4. The relatively low number of seroconversions indicates a low but measurable risk of exposure to human filarial parasites among Australian soldiers deployed to Timor-Leste. However, to reduce the risk of exposure to these parasites, soldiers deploying to endemic areas should practice strict adherence to personal protective measures against mosquito bites. PMID- 18385350 TI - Oxidative and nitrative stress in Opisthorchis viverrini-infected hamsters: an indirect effect after praziquantel treatment. AB - Praziquantel causes adverse effects after short-term treatment. To examine the mechanism of these effects, we studied the distribution of Opisthorchis viverrini antigens and the expression of inducible nitric oxide synthase (iNOS), nuclear factor-kappaB (NF-kappaB), and antioxidant enzymes in O. viverrini-infected hamsters during short-term praziquantel treatment. Praziquantel-induced dispersion of parasite antigens produced a recruitment of inflammatory cells. NF kappaB and iNOS mRNA expression was significantly elevated and associated with their immunoreactivity in the bile duct epithelium and inflammatory cells. Plasma nitrate, end products of nitric oxide, and malondialdehyde level increased significantly. Expression of mRNA for antioxidant enzymes (superoxide dismutases, catalase, and glutathione peroxidase) also increased significantly, which suggests host defense against oxidative stress. These results suggest that short term praziquantel treatment induces inflammation and resulting oxidative and nitrative stress through O. viverrini antigen release. Data in this study can be used as a basis to understand potential side effects of praziquantel treatment in humans. PMID- 18385351 TI - Helicobacter pylori seroprevalence in Amerindians from isolated locations. AB - Helicobacter pylori seems universally distributed in all human populations, with high prevalence in the third world. Because H. pylori is an ancestral indigenous microbe of the human stomach, we hypothesized that its prevalence in isolated Amerindians would be high. A serologic study was performed on 19 Guahibo-Piaroa and 17 Warao in Venezuela, using H. pylori whole cell (WC) and CagA antigens from US strains. For Guahibo-Piaroa Amerindians, CagA seropositivity was 95%, but WC seropositivity was only 74%. For Warao, both CagA and WC seropositive proportions were low (65% and 76%, respectively). Because all CagA-seropositive individuals carry H. pylori, the results suggest that there has been bacterial antigen divergence, probably caused by genetic drift/natural selection, on humans and their microbes in isolated human groups. PMID- 18385352 TI - PCR detection of Clostridium difficile triose phosphate isomerase (tpi), toxin A (tcdA), toxin B (tcdB), binary toxin (cdtA, cdtB), and tcdC genes in Vhembe District, South Africa. AB - Specific polymerase chain reaction (PCR) protocols were used to determine the prevalence of toxigenic Clostridium difficile in Vhembe, South Africa. Of 322 stool samples collected, toxigenic C. difficile was found in 23 (7.1%) cases and was significantly associated with diarrhea 20 (11.4%) compared with 3 (2%) in non diarrheal samples (chi(2) = 426, P = 0.001), intestinal inflammation in 18 (12.1%) compared with 5 (2.9%) in lactoferrin-negative samples (chi(2) = 10.194, P = 0.001), and occult blood in 19 (16%) compared with 4 (2%) in occult blood negative samples (chi(2) = 22.157, P < 0.001). Toxigenic C. difficile was more common among individuals > 50 years of age (20%), followed by those between 30 and 39 years of age (19%) and was not associated with HIV infections (chi(2) = 0.289, P = 0.591). Co-infection with other pathogens was common. Multivariate analysis indicated that toxigenic C. difficile was associated with E. bieneusi (P = 0.028), C. parvum (P = 0.007), and Enteroaggregative Escherichia coli (EAEC) (P = 0.007) in diarrheal samples. This study confirms the usefulness of PCR methodologies in the detection of toxigenic C. difficile and suggests that C. difficile is responsible for a small, but underappreciated, proportion of diarrheal cases in the region, and further study is warranted in this area. PMID- 18385353 TI - Ecoepidemiology of tularemia in the southcentral United States. AB - We combined county-based data for tularemia incidence from 1990 to 2003 for a nine-state region (Arkansas, Illinois, Indiana, Kansas, Kentucky, Missouri, Nebraska, Oklahoma, and Tennessee) in the southcentral United States with Geographic Information System (GIS)-based environmental data to determine associations between coverage by different habitats (especially dry forest representing suitable tick habitat) and tularemia incidence. High-risk counties (> 1 case per 100,000 person-years) clustered in Arkansas-Missouri and far eastern Oklahoma and Kansas. County tularemia incidence was positively associated with coverage by dry forested habitat suitable for vector ticks for Oklahoma Kansas-Nebraska and Arkansas-Missouri but not for Illinois-Indiana-Kentucky Tennessee. A multivariate logistic regression model predicting presence of areas with risk of tularemia based on GIS-derived environmental data was developed for the Arkansas-Missouri tularemia focus. The study shows the potential for research on tularemia ecoepidemiology and highlights the need for further modeling efforts based on acarologic data and more fine-scale point or zip code/census tract epidemiologic data. PMID- 18385355 TI - Expanded range of Burkholderia species in Australia. AB - This study describes the isolation and characterization of several Burkholderia species from soil in northern Australia. Phenotypic and molecular tests indicate that these isolates belong to the species Burkholderia thailandensis and Burkholderia ubonensis. These observations significantly extend our knowledge of the geographic distribution of these 2 species. Evidence of these species in Australia has implications for bacterial identification in clinical laboratories, diagnostic serology tests, and environmental biodiversity studies. PMID- 18385354 TI - Brucellosis in household members of Brucella patients residing in a large urban setting in Peru. AB - During home visits and using a point-of-care test for brucellosis, we screened the household members of adult patients found to have brucellosis by investigation at the Hospital Nacional Daniel Alcides Carrion in Callao, Peru. A total of 206 household members of 43 patients were screened, and 15 (7.3%) household members in 10 (23.3%) households tested seropositive. Brucellosis was diagnosed in 14 of them, all but 4 presenting with acute or subacute uncomplicated disease. Regardless of attempts to control brucellosis in Peru, the disease continues to be reasonably common among household members of brucellosis patients. Household members presumably remain the single most important identifiable risk group in an urban setting, and screening them provides an effective means for their early diagnosis. Although contact with livestock was rare, the consumption of unpasteurized dairy products was reported by almost all patients with brucellosis, their household members, and hospitalized non brucellosis patients. PMID- 18385356 TI - The functional state of the complement system in leprosy. AB - Ninety-one patients with different clinical forms of leprosy, 36 lepromatous (LL), 33 tuberculoid (TL), and 22 dimorphic (DL), and 31 healthy volunteer donors were included in this study. Total complement system (CS) activity was assessed by hemolytic methods, whereas individual components were quantified by the enzyme linked immunosorbent assay. Under conditions allowing initiation of cascade by the classic pathway (CP) but not alternative pathway (AP) activation, significant CS consumption was detected only in sera from patients with LL. In this group of patients, C4 but not factor B (fB) or C3 was significantly reduced, whereas mannose-binding lectin (MBL) serum levels were significantly higher. These results indicate that the CP is involved in CS activation in patients infected with Mycobacterium leprae manifesting LL clinical form of leprosy. An association is likely between circulating immune complexes and MBL high serum levels for initiation of CS activation in patients with LL form of leprosy. PMID- 18385357 TI - Multiple modes of transmission of giardiasis in primary schoolchildren of a rural community, Thailand. AB - In February 2005, we conducted a cross-sectional study to determine the prevalence and the risk factors of giardiasis in 531 primary schoolchildren of a rural community, Chacheongsao province, Thailand. Using both sedimentation and flotation techniques to detect Giardia duodenalis, the prevalence of giardiasis was 6.2%. Assemblage A, subgenotype II and assemblage B, subgenotype IV were identified by PCR-RFLP of glutamate dehydrogenase gene. Our data might indicate that, in this population, only assemblage A, subgenotype II of G. duodenalis was transmitted via water. Using multivariate analysis, significant risk factors for giardiasis were children of age 5-9 years, households with > or = 3 children under the age of 12 years, low parental educational level, drinking bottled water, and living in close contact with dogs. Washing hands before meals had a protective effect. From these significant risk factors, multiple modes of transmission of G. duodenalis were suggested in this population. PMID- 18385358 TI - A novel exo-antigen-based ELISA for the detection of canine leishmaniasis. AB - Dogs which are infected with leishmania parasites serve as major reservoir hosts for zoonotic visceral leishmaniasis. The incidence of zoonotic visceral leishmaniasis is rising in many countries. This may be associated with the continuing drift of people and their pets from rural areas into peri-urban settings, particularly at the fringe of large cities. At the same time, there is evidence of adaptation of sand fly vectors to these urban settings. This has created an alarming situation because, even though domestic and stray dogs may be infected, many remain asymptomatic but are still highly infectious to the sand fly vectors and thus pose a serious threat to human health. Over half of the infected dogs have asymptomatic infections and current assays are not sensitive enough under field conditions to distinguish asymptomatic from symptomatic dogs. There is an urgent need for a specific and sensitive screening tool for use in the field. We have previously demonstrated that promastigote exo-antigen-based ELISAs can be used in the specific diagnosis of human visceral leishmaniasis (HVL). A cocktail of exo-antigens prepared from three species (L. infantum, L. donovani, and L. major) was used to develop and optimize a canine ELISA assay. Serum samples from dogs with a variety of pathological conditions but living in a non-leishmania endemic area were used as negative controls and their reactivity was used to determine a cut-off value for the ELISA. Samples from dogs residing in a leishmania endemic area were tested in parallel using direct agglutination (DAT), immunofluorescence (IFAT), and ELISA. The ELISA results correlated closely (100%) with the clinical symptoms, and were elevated in one asymptomatic dog. This sample was also found to be positive by IFAT. Based on its sensitivity and specificity, the cocktail exo-antigen-based ELISA may prove useful, even at 1:2,000 serum dilutions, for screening dogs in different geographical regions of the world. PMID- 18385359 TI - The value of molecular studies for the diagnosis of congenital Chagas disease in northeastern Argentina. AB - Polymerase chain reaction (PCR) as a diagnostic tool for congenital Chagas disease is scarcely used. To assess the usefulness of PCR, a study in neonates was carried out to compare PCR with standard methods; that is bloodstream parasites by microhematocrit, and/or seroconversion by two conventional procedures. Seventeen neonates, born to mothers with positive Trypanosoma cruzi serology were analyzed by serologic, parasitological and molecular procedures, in blood samples taken at birth, and when aged 2-4 and 9 months. For a better comparison of sensitivity between both parasitological methods (microhematocrit and PCR), the number of samples of neonates studied at birth extended to 121. In children followed-up until 9 months of age, a 100% correlation between PCR and routine diagnosis was found, with PCR showing its highest sensitivity from Month 2 onwards. Comparative analysis between both parasitological methods, on samples taken at birth, showed a higher sensitivity of PCR respect the microhematocrit (P < 0.0008). PMID- 18385360 TI - Evaluation of a modified pulsed-field gel electrophoresis approach for the identification of Leptospira serovars. AB - The genus Leptospira is composed of spirochetes that are morphologically indistinguishable and cannot be differentiated by phenotypic methods. Determining the identity of an infecting serovar is valuable from both epidemiologic and public health standpoints. Pulsed-field gel electrophoresis (PFGE) of Leptospira has had limited use in few laboratories. In this study, we modified the existing PFGE protocol to reduce time to completion and developed a PFGE database using the restriction endonuclease Not I to generate PFGE profiles for the identification and evaluation of Leptospira. Reference strains (n = 206) from a collection maintained by the Centers for Disease Control and Prevention were studied. Eighty-nine percent of the serovars produced distinct patterns, and different strains of the same serovar showed profiles that were highly similar. Pulsed-field gel electrophoresis of Leptospira is a useful tool for molecular characterization of reference serovars, the investigation of potentially new species or serovars, and ultimately for the routine identification of clinical isolates. PMID- 18385361 TI - Association of pfcrt but not pfmdr1 alleles with chloroquine resistance in Iranian isolates of Plasmodium falciparum. AB - This study was designed to analyze the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and P. falciparum multidrug resistance 1 (pfmdr1) mutations as markers of chloroquine (CQ) resistance in 200 blood samples collected from malaria patients in south-eastern Iran during 2002-2005. Among these, 25 (post-treatment) fulfilled the 28-day follow-up study. A high number of Iranian P. falciparum (97%) strains harbored quadruple mutations at codons 76T, 220S, 326D, and 356L. All post-treatment isolates harbored the mutant allele 76T, but low rates of the mutant allele 86Y (44%) of the pfmdr1 gene were detected. No wild haplotype of pfcrt (72-CVMNKAQNIR-371) was found in post-treatment samples; however, 56% of clinical "failure" samples carried the wild type of pfmdr1 (NYSND). The present results suggest a strong association between pfcrt 76T, but not pfmdr1 86Y mutation and in vivo CQ resistance. Furthermore, we found the CQ resistance-associated SVMNT haplotype, which previously had been seen in South American isolates. Although Iran is located more proximally to Southeast Asia than to South America, no CQ resistance-associated CVIET haplotye has been observed in this region. Therefore, these results were not consistent with the earlier presumed spread of CQR parasites from Southeast Asia to Africa via the Indian subcontinent. In conclusion, P. falciparum mutations associated with resistance to CQ are abundant in south-eastern Iran and this finding strongly supports that CQ as the first line drug is inadequate for treatment of uncomplicated falciparum malaria in Iran. PMID- 18385362 TI - Pharmacokinetics of dihydroartemisinin in a murine malaria model. AB - Pharmacokinetic properties of dihydroartemisinin (DHA) were determined in mice given 100 mg/kg intraperitoneal DHA. Half-life, CL/F, and V/F were 25 min, 61.3 L/hr/kg, and 36.3 L/kg in malaria-infected mice and 19 min, 50.9 L/hr/kg, and 23.0 L/kg in controls. These data are valuable for pharmacokinetic pharmacodynamic evaluations of DHA in murine models. PMID- 18385363 TI - Detection of new Babesia microti-like parasites in a rhesus monkey (Macaca mulatta) with a suppressed Plasmodium cynomolgi infection. AB - A new type of piroplasm, phylogenetically closest to Babesia microti-like parasites previously detected in Eurasian red squirrels (Sciurus vulgaris orientis), was identified in a rhesus monkey (Macaca mulatta) imported from China. After challenge with Plasmodium cynomolgi M strain blood-stage parasites, the rhesus monkey repeatedly showed markedly reduced levels of Plasmodium parasitemia when compared with animals not infected with this organism. PMID- 18385364 TI - Malaria and Hepatocystis species in wild macaques, southern Thailand. AB - Southeast Asian macaques are natural hosts for a number of nonhuman primate malaria parasites; some of these can cause diseases in humans. We conducted a cross-sectional survey by collecting 99 blood samples from Macaca fascicularis in southern Thailand. Giemsa-stained blood films showed five (5.1%) positive samples and six (6.1%) isolates had positive test results by polymerase chain reaction. A phylogenetic tree inferred from the A-type sequences of the small subunit ribosomal RNA gene confirmed Plasmodium inui in five macaques; one of these macaques was co-infected with P. coatneyi. Hepatocystis, a hemoprotozoan parasite transmitted by Culicoides, was identified in an isolate that was confirmed by analysis of mitochondrial cytochrome b sequences. All malaria-infected monkeys lived in mangrove forests, but no infected monkeys were found in an urban area. These findings indicate regional differences in malaria distribution among these macaques, as well as differences in potential risk of disease transmission to humans. PMID- 18385365 TI - Combining mosquito vector and human disease data for improved assessment of spatial West Nile virus disease risk. AB - Assessments of spatial risk of exposure to vector-borne pathogens that combine vector and human disease data are needed for areas encompassing large tracts of public land with low population bases. We addressed this need for West Nile virus (WNV) disease in the northern Colorado Front Range by developing not only a spatial model for entomological risk of exposure to Culex tarsalis WNV vectors and an epidemiological risk map for WNV disease but also a novel risk classification index combining data for these independently derived measures of entomological and epidemiological risk. Risk of vector exposure was high in the densely populated eastern plains portion of the Front Range but low in cooler montane areas to the west that are sparsely populated but used heavily for recreation in the summer. The entomological risk model performed well when applied to the western, mountainous part of Colorado and validated against epidemiologic data. PMID- 18385366 TI - First isolation of West Nile virus in the Caribbean. AB - A sentinel chicken program for West Nile virus (WNV) surveillance was initiated in July 2006 in eastern Puerto Rico, yielding the first seroconversions on June 4, 2007. WNV was isolated from sentinel chicken serum and mosquito pools (Culex nigripalpus, Culex bahamensis) for the first time in Tropical America. Preliminary sequence analysis of the prM and E genes revealed a 1-amino acid difference (V159A) between the Puerto Rican 2007 and the NY99. This mutation has been observed in the current dominant clade circulating in the United States. Sentinel chicken surveillance was a useful tool for the detection of West Nile virus in the tropics. PMID- 18385367 TI - Choclo virus infection in the Syrian golden hamster. AB - Andes virus and Choclo virus are agents of hantavirus pulmonary syndrome. Andes virus in hamsters almost always causes a disease that is pathologically indistinguishable from fatal hantavirus pulmonary syndrome. The purpose of this study was to assess the pathogenicity of Choclo virus in hamsters. None of 18 hamsters infected with Choclo virus exhibited any symptom of disease. No evidence of inflammation or edema was found in the lungs of the 10 animals killed on days 7, 9, 11, 13, and 16 post-inoculation or in the lungs of the 8 animals killed on day 28 post-inoculation; however, hantavirus antigen was present in large numbers of endothelial cells in the microvasculature of the lungs of the animals killed on days 7, 9, 11, and 13 post-inoculation. These results suggest that infection in the microvasculature of lung tissue alone does not result in the life threatening pulmonary edema in hamsters infected with Andes virus. PMID- 18385368 TI - Dhori virus (Orthomyxoviridae: Thogotovirus) infection of mice produces a disease and cytokine response pattern similar to that of highly virulent influenza A (H5N1) virus infection in humans. AB - Mice infected with Dhori virus (DHOV) develop a fulminant, systemic, and uniformly fatal illness that has many of the clinical and pathologic findings seen in H5N1 influenza A virus infection. However, the role of host's immune response in DHOV infection remains unclear. In this study, the concentrations of 23 inflammatory cytokines and chemokines were measured in the liver, lungs, and sera of mice during the course of DHOV infection. Liver function, level of viremia, and hematologic response were also monitored. Infected animals exhibited significant leucopenia and lymphopenia, which directly correlated with the disease progression. High yields of infectious virus along with strikingly elevated expression of various inflammatory mediators, including tumor necrosis factor (TNF)-alpha, interleukin (IL)-1, IL-6, IL-10, macrophage inflammatory protein (MIP)-1alpha, manocyte chemoattractant protein (MCP)-1, and interferon (IFN)-alpha, indicate that these responses play an important role in the observed disease and pathology. The overall clinical, pathologic, and immunologic responses of ICR mice to DHOV infection closely resemble those described for highly virulent influenza A virus infection in humans, thereby offering a realistic, safe, and alternative animal model for studying the pathogenesis and treatment of highly pathogenic avian influenza virus. PMID- 18385369 TI - Limited interdecadal variation in mosquito (Diptera: Culicidae) and avian host competence for Western equine encephalomyelitis virus (Togaviridae: Alphavirus). AB - Historically, western equine encephalomyelitis virus (WEEV) caused large equine and human epidemics in the Americas from Canada into Argentina. Despite recent enhanced surveillance for West Nile virus, there have been few reports of equine or human cases and little documented enzootic activity of WEEV. During the past three years, WEEV has been active again in California, but without human or equine cases. In the current study, we compared host and vector competence of representative WEEV isolates made during each decade over the past 60 years using white-crowned sparrows, house sparrows, and Culex tarsalis Coquillett as representative hosts. Results indicated limited time-related change in virulence among WEEV strains in birds and little difference in vector competence in Cx. tarsalis. Although temporal and spatial genetic changes have been documented, these seem to present limited phenotypic change in host competence and cannot explain the absence of equine and human cases. PMID- 18385370 TI - Rocky Mountain spotted fever in Argentina. AB - We describe the first molecular confirmation of Rickettsia rickettsii, the cause of Rocky Mountain spotted fever (RMSF), from a tick vector, Amblyomma cajennense, and from a cluster of fatal spotted fever cases in Argentina. Questing A. cajennense ticks were collected at or near sites of presumed or confirmed cases of spotted fever rickettsiosis in Jujuy Province and evaluated by polymerase chain reaction assays for spotted fever group rickettsiae. DNA of R. rickettsii was amplified from a pool of A. cajennense ticks and from tissues of one of four patients who died during 2003-2004 after illnesses characterized by high fever, severe headache, myalgias, and petechial rash. The diagnosis of spotted fever rickettsiosis was confirmed in the other patients by indirect immunofluorescence antibody and immunohistochemical staining techniques. These findings show the existence of RMSF in Argentina and emphasize the need for clinicians throughout the Americas to consider RMSF in patients with febrile rash illnesses. PMID- 18385372 TI - A breakthrough innovation in animal evolution. PMID- 18385371 TI - Dicer loss in striatal neurons produces behavioral and neuroanatomical phenotypes in the absence of neurodegeneration. AB - MicroRNAs (miRNAs) are small noncoding RNAs that can act to repress target mRNAs by suppressing translation and/or reducing mRNA stability. Although it is clear that miRNAs and Dicer, an RNase III enzyme that is central to the production of mature miRNAs, have a role in the early development of neurons, their roles in the postmitotic neuron in vivo are largely unknown. To determine the roles of Dicer in neurons, we ablated Dicer in dopaminoceptive neurons. Mice that have lost Dicer in these cells display a range of phenotypes including ataxia, front and hind limb clasping, reduced brain size, and smaller neurons. Surprisingly, dopaminoceptive neurons without Dicer survive over the life of the animal. The lack of profound cell death contrasts with other mouse models in which Dicer has been ablated. These studies highlight the complicated nature of Dicer ablation in the brain and provide a useful mouse model for studying dopaminoceptive neuron function. PMID- 18385373 TI - Postendocytotic traffic of the galanin R1 receptor: a lysosomal signal motif on the cytoplasmic terminus. AB - The neuropeptide galanin R1 receptor (GalR1) was tagged at its C terminus with EGFP (GalR1-EGFP) to study receptor localization and trafficking. In PC12 and HEK293 cells, functional GalR1-EGFP was expressed on the plasma membrane and internalized into cytoplasmic vesicles after galanin stimulation. The internalization was blocked by 0.4 M sucrose and by silencing of clathrin with siRNA methodology. Internalized GalR1-EGFP and LysoTracker, a lysosomal marker, overlapped in intracellular vesicles after prolonged galanin stimulation. This colocalization was strongly reduced after site-directed mutagenesis of the motif YXXO on the C terminus of GalR1 (where O is a bulky hydrophobic residue and X any amino acid). Taken together, these data suggest that GalR1 is internalized via the clathrin-dependent, endocytic pathway and then, to a large extent, delivered to lysosomes for degradation through the lysosome-targeting signal YXXO. PMID- 18385374 TI - Regulation of polymerase exchange between Poleta and Poldelta by monoubiquitination of PCNA and the movement of DNA polymerase holoenzyme. AB - To ensure efficient and timely replication of genomic DNA, organisms in all three kingdoms of life possess specialized translesion DNA synthesis (TLS) polymerases (Pols) that tolerate various types of DNA lesions. It has been proposed that an exchange between the replicative DNA Pol and the TLS Pol at the site of DNA damage enables lesion bypass to occur. However, to date the molecular mechanism underlying this process is not fully understood. In this study, we demonstrated in a reconstituted system that the exchange of Saccharomyces cerevisiae Poldelta with Poleta requires both the stalling of the holoenzyme and the monoubiquitination of proliferating cell nuclear antigen (PCNA). A moving Poldelta holoenzyme is refractory to the incoming Poleta. Furthermore, we showed that the Poleta C-terminal PCNA-interacting protein motif is required for the exchange process. We also demonstrated that the second exchange step to bring back Poldelta is prohibited when Lys-164 of PCNA is monoubiquitinated. Thus the removal of the ubiquitin moiety from PCNA is likely required for the reverse exchange step after the lesion bypass synthesis by Poleta. PMID- 18385375 TI - Aminoacyl-tRNAs, the bacterial cell envelope, and antibiotics. PMID- 18385376 TI - Purported nanobacteria in human blood as calcium carbonate nanoparticles. AB - Recent evidence suggests a role for nanobacteria in a growing number of human diseases, including renal stone formation, cardiovascular diseases, and cancer. This large body of research studies promotes the view that nanobacteria are not only alive but that they are associated with disease pathogenesis. However, it is still unclear whether they represent novel life forms, overlooked nanometer-size bacteria, or some other primitive self-replicating microorganisms. Here, we report that CaCO(3) precipitates prepared in vitro are remarkably similar to purported nanobacteria in terms of their uniformly sized, membrane-delineated vesicular shapes, with cellular division-like formations and aggregations in the form of colonies. The gradual appearance of nanobacteria-like particles in incubated human serum as well as the changes seen with their size and shape can be influenced and explained by introducing varying levels of CO(2) and NaHCO(3) as well as other conditions known to influence the precipitation of CaCO(3). Western blotting reveals that the monoclonal antibodies, claimed to be specific for nanobacteria, react in fact with serum albumin. Furthermore, nanobacteria like particles obtained from human blood are able to withstand high doses of gamma-irradiation up to 30 kGy, and no bacterial DNA is found by performing broad range PCR amplifications. Collectively, our results provide a more plausible abiotic explanation for the unusual properties of purported nanobacteria. PMID- 18385377 TI - Molecular links among the causative genes for ocular malformation: Otx2 and Sox2 coregulate Rax expression. AB - The neural-related genes Sox2, Pax6, Otx2, and Rax have been associated with severe ocular malformations such as anophthalmia and microphthalmia, but it remains unclear as to how these genes are linked functionally. We analyzed the upstream signaling of Xenopus Rax (also known as Rx1) and identified the Otx2 and Sox2 proteins as direct upstream regulators of Rax. We revealed that endogenous Otx2 and Sox2 proteins bound to the conserved noncoding sequence (CNS1) located approximately 2 kb upstream of the Rax promoter. This sequence is conserved among vertebrates and is required for potent transcriptional activity. Reporter assays showed that Otx2 and Sox2 synergistically activated transcription via CNS1. Furthermore, the Otx2 and Sox2 proteins physically interacted with each other, and this interaction was affected by the Sox2-missense mutations identified in these ocular disorders. These results demonstrate that the direct interaction and interdependence between the Otx2 and Sox2 proteins coordinate Rax expression in eye development, providing molecular linkages among the genes responsible for ocular malformation. PMID- 18385378 TI - Alteration of BACE1-dependent NRG1/ErbB4 signaling and schizophrenia-like phenotypes in BACE1-null mice. AB - beta-Site APP-cleaving enzyme 1 (BACE1) is required for the penultimate cleavage of the amyloid-beta precursor protein (APP) leading to the generation of amyloid beta peptides that is central to the pathogenesis of Alzheimer's disease. In addition to its role in endoproteolysis of APP, BACE1 participates in the proteolytic processing of neuregulin 1 (NRG1) and influences the myelination of central and peripheral axons. Although NRG1 has been genetically linked to schizophrenia and NRG1(+/-) mice exhibit a number of schizophrenia-like behavioral traits, it is not known whether altered BACE1-dependent NRG1 signaling can cause similar behavioral abnormalities. To test this hypothesis, we analyze the behaviors considered to be rodent analogs of clinical features of schizophrenia in BACE1(-/-) mice with impaired processing of NRG1. We demonstrate that BACE1(-/-) mice exhibit deficits in prepulse inhibition, novelty-induced hyperactivity, hypersensitivity to a glutamatergic psychostimulant (MK-801), cognitive impairments, and deficits in social recognition. Importantly, some of these manifestations were responsive to treatment with clozapine, an atypical antipsychotic drug. Moreover, although the total amount of ErbB4, a receptor for NRG1 was not changed, binding of ErbB4 with postsynaptic density protein 95 (PSD95) was significantly reduced in the brains of BACE1(-/-) mice. Consistent with the role of ErbB4 in spine morphology and synaptic function, BACE1(-/-) mice displayed reduced spine density in hippocampal pyramidal neurons. Collectively, our findings suggest that alterations in BACE1-dependent NRG1/ErbB4 signaling may participate in the pathogenesis of schizophrenia and related psychiatric disorders. PMID- 18385379 TI - Serotonin is necessary for place memory in Drosophila. AB - Biogenic amines, such as serotonin and dopamine, can be important in reinforcing associative learning. This function is evident as changes in memory performance with manipulation of either of these signals. In the insects, evidence begins to argue for a common role of dopamine in negatively reinforced memory. In contrast, the role of the serotonergic system in reinforcing insect associative learning is either unclear or controversial. We investigated the role of both of these signals in operant place learning in Drosophila. By genetically altering serotonin and dopamine levels, manipulating the neurons that make serotonin and dopamine, and pharmacological treatments we provide clear evidence that serotonin, but not dopamine, is necessary for place memory. Thus, serotonin can be critical for memory formation in an insect, and dopamine is not a universal negatively reinforcing signal. PMID- 18385380 TI - The thermal impulse response of Escherichia coli. AB - Swimming Escherichia coli responds to changes in temperature by modifying its motor behavior. Previous studies using populations of cells have shown that E. coli accumulate in spatial thermal gradients, but these experiments did not cleanly separate thermal responses from chemotactic responses. Here we have isolated the thermal response by studying the behavior of single, tethered cells. The motor output of cells grown at 33 degrees C was measured at constant temperature, from 10 degrees to 40 degrees C, and in response to small, impulsive increases in temperature, from 23 degrees to 43 degrees C. The thermal impulse response at temperatures < 31 degrees C is similar to the chemotactic impulse response: Both follow a similar time course, share the same directionality, and show biphasic characteristics. At temperatures > 31 degrees C, some cells show an inverted response, switching from warm- to cold-seeking behavior. The fraction of inverted responses increases nonlinearly with temperature, switching steeply at the preferred temperature of 37 degrees C. PMID- 18385381 TI - The role of filament-packing dynamics in powering amoeboid cell motility. AB - Although several models have been proposed to account for how cytoskeleton polymerization drives protrusion in cell motility, the precise mechanism remains controversial. Here, we show that, in addition to force exerted directly against the membrane by growing filaments, the way elongating filaments pack also contributes to protrusion by generating an expansion of the cytoskeleton gel. Tomography shows that filament packing in the major sperm protein (MSP) -based nematode sperm-motility machinery resembles that observed with rigid rods. Maximum rod-packing density decreases dramatically as the rods lengthen. Therefore, as filaments elongate, the cytoskeleton gel expands to accommodate their packing less densely. This volume expansion combines with polymerization to drive protrusion. Consistent with this hypothesis, an engineered MSP mutant that generates shorter filaments shows higher filament-packing density and slower movement. PMID- 18385382 TI - Touch perception reveals the dominance of spatial over digital representation of numbers. AB - We learn counting on our fingers, and the digital representation of numbers we develop is still present in adulthood [Andres M, et al. (2007) J Cognit Neurosci 19:563-576]. Such an anatomy-magnitude association establishes tight functional correspondences between fingers and numbers [Di Luca S, et al. (2006) Q J Exp Psychol 59:1648-1663]. However, it has long been known that small-to-large magnitude information is arranged left-to-right along a mental number line [Dehaene S, et al. (1993) J Exp Psychol Genet 122:371-396]. Here, we investigated touch perception to disambiguate whether number representation is embodied on the hand ("1" = thumb; "5" = little finger) or disembodied in the extrapersonal space ("1" = left; "5" = right). We directly contrasted these number representations in two experiments using a single centrally located effector (the foot) and a simple postural manipulation of the hand (palm-up vs. palm-down). We show that visual presentation of a number ("1" or "5") shifts attention cross-modally, modulating the detection of tactile stimuli delivered on the little finger or thumb. With the hand resting palm-down, subjects perform better when reporting tactile stimuli delivered to the little finger after presentation of number "5" than number "1." Crucially, this pattern reverses (better performance after number "1" than "5") when the hand is in a palm-up posture, in which the position of the fingers in external space, but not their relative anatomical position, is reversed. The human brain can thus use either space- or body-based representation of numbers, but in case of competition, the former dominates the latter, showing the stronger role played by the mental number line organization. PMID- 18385383 TI - Killin is a p53-regulated nuclear inhibitor of DNA synthesis. AB - Cell growth arrest and apoptosis are two best-known biological functions of tumor suppressor p53. However, genetic evidence indicates that not only is p21 the major mediator of G(1) arrest, but also it can prevent apoptosis with an unknown mechanism. Here, we report the discovery of a p53 target gene dubbed killin, which lies in close proximity to pten on human chromosome 10 and encodes a 20-kDa nuclear protein. We show that Killin is not only necessary but also sufficient for p53-induced apoptosis. Genetic and biochemical analysis demonstrates that Killin is a high-affinity DNA-binding protein, which potently inhibits eukaryotic DNA synthesis in vitro and appears to trigger S phase arrest before apoptosis in vivo. The DNA-binding domain essential for DNA synthesis inhibition was mapped to within 42 amino acid residues near the N terminus of Killin. These results support Killin as a missing link between p53 activation and S phase checkpoint control designed to eliminate replicating precancerous cells, should they escape G(1) blockade mediated by p21. PMID- 18385385 TI - Patterns of interstitial inflammation during the evolution of renal injury in experimental aristolochic acid nephropathy. AB - BACKGROUND: Interstitial inflammation is a prominent feature associated with the severity of renal injury and progressive kidney failure. We utilized an animal model of aristolochic acid (AA)-induced nephropathy (AAN) to assess patterns of infiltration and inflammation during the evolution of tubulointerstitial damage and to relate them to the development of fibrosis. METHODS: Male Wistar rats receiving sc daily AA or vehicle were sacrificed between Days 1 and 35. Infiltrating mononuclear cells were characterized by immunohistochemistry. The kidney infiltrating T lymphocytes were phenotyped by flow cytometry. Urinary levels of Th-1/ Th-2 cytokines, of monocyte chemoattractant protein-1 and of active transforming growth factor-beta (TGF-beta) were measured. Tissue expression of phosphorylated smad 2/3 protein was used to examine the TGF-beta signalling pathway. RESULTS: In AA rats, monocytes/macrophages and T lymphocytes predominantly infiltrated areas of necrotic proximal tubular cells. The coexpressions of ED1 and/or Ki-67/MHCII by infiltrating cells reflected monocyte/macrophage proliferation and their activation, respectively. The accumulation of cytotoxic T lymphocytes was attested by severe signs of CD8+ cell tubulitis. The CD8/E-cadherin costaining confirmed intrarenal homing of CD8+CD103+ cells. Urinary levels of proinflammatory cytokines and of active TGF beta significantly increased at Days 10 and 35. An early and persistent nuclear overexpression of phosphorylated smad 2/3 protein was detected in tubular and interstitial compartments. CONCLUSION: An early and massive interstitial inflammation characterized by activated monocytes/macrophages and cytotoxic CD8+CD103+ T lymphocytes is demonstrated for the first time during the progression of experimental AAN. The involvement in an interstitial fibrosis onset of active TGF-beta is highly suggested, at least via the psmad 2/3 intracellular signalling pathway. PMID- 18385386 TI - Chronic allograft nephropathy--clinical guidance for early detection and early intervention strategies. PMID- 18385384 TI - Detecting evolutionary relationships across existing fold space, using sequence order-independent profile-profile alignments. AB - Here, a scalable, accurate, reliable, and robust protein functional site comparison algorithm is presented. The key components of the algorithm consist of a reduced representation of the protein structure and a sequence order independent profile-profile alignment (SOIPPA). We show that SOIPPA is able to detect distant evolutionary relationships in cases where both a global sequence and structure relationship remains obscure. Results suggest evolutionary relationships across several previously evolutionary distinct protein structure superfamilies. SOIPPA, along with an increased coverage of protein fold space afforded by the structural genomics initiative, can be used to further test the notion that fold space is continuous rather than discrete. PMID- 18385387 TI - Understanding renal disorders as systemic diseases: the fascinating world of basement membranes beyond the glomerulus. PMID- 18385388 TI - Rituximab in membranous nephropathy after haematopoietic stem cell transplantation. PMID- 18385389 TI - The molecular mechanisms of the attenuation of cisplatin-induced acute renal failure by N-acetylcysteine in rats. AB - BACKGROUND: The clinical use of cisplatin (cis-diamminedichloro-platinum II, CDDP) is highly limited by its nephrotoxicity. Although N-acetylcysteine (NAC), a thiol-containing antioxidant, has been documented to be effective in attenuating renal injury induced by CDDP, the precise mechanisms involved in its renoprotection have not been completely clarified. METHODS: We investigated the effects of NAC on oxidative stress and oxidation-associated signals, such as p38 mitogen-activated protein kinase (MAPK), NF-kappaB and TNF-alpha, in CDDP-induced acute renal failure (ARF) rats, in comparison to the effects of melatonin (MT), one of the physiological TNF-alpha inhibitors, and pyrrolidine dithiocarbamate (PDTC), a NF-kappaB inhibitor. RESULTS: NAC blocked oxidative stress, p38 MAPK activation, caspase-3 cleavage, tissue apoptosis, renal dysfunction and morphological damage induced by CDDP. CDDP-triggered NF-kappaB translocation into the nucleus and TNF-alpha mRNA increase in the kidney were also inhibited in NAC treated rats. MT downregulated the TNF-alpha mRNA level, and PDTC inhibited the increases in both NF-kappaB translocation and TNF-alpha mRNA. Neither MT nor PDTC were capable of interfering with oxidative stress, p38 MAPK phosphorylation, caspase-3 cleavage, tissue apoptosis and kidney injury induced by CDDP. CONCLUSIONS: These data suggest that oxidative stress and p38 MAPK-mediated apoptotic cell death pathways are involved, at least in part, in the pathogenesis of CDDP-induced ARF, and negative regulation of p38 MAPK activation through inhibition of oxidative stress appears to play a central role in the beneficial effects of NAC. PMID- 18385391 TI - A randomized, double-blind, placebo-controlled trial of niacinamide for reduction of phosphorus in hemodialysis patients. AB - BACKGROUND AND OBJECTIVES: Niacinamide inhibits intestinal sodium/phosphorus transporters and reduces serum phosphorus in open-label studies. A prospective, randomized, double-blind, placebo-controlled crossover trial was performed for assessment of the safety and efficacy of niacinamide. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Hemodialysis patients with phosphorus levels > or =5.0 mg/dl were randomly assigned to 8 wk of niacinamide or placebo, titrated from 500 to 1500 mg/d. After a 2-wk washout period, patients switched to 8 wk of the alternative therapy. Vitamin D analogs and calcimimetics were held constant; phosphorus binders were not changed unless safety criteria were met. RESULTS: Thirty-three patients successfully completed the trial. Serum phosphorus fell significantly from 6.26 to 5.47 mg/dl with niacinamide but not with placebo (5.85 to 5.98 mg/dl). A concurrent fall in calcium-phosphorus product was seen with niacinamide, whereas serum calcium, intact parathyroid hormone, uric acid, platelet, triglyceride, LDL, and total cholesterol levels remained stable in both arms. Serum HDL levels rose with niacinamide (50 to 61 mg/dl but not with placebo. Adverse effects were similar between both groups. Among patients who were > or =80% compliant, results were similar, although the decrease in serum phosphorus with niacinamide was more pronounced (6.45 to 5.28 mg/dl) and the increase in HDL approached significance (49 to 58 mg/dl). CONCLUSIONS: In hemodialysis patients, niacinamide effectively reduces serum phosphorus when co administered with binders and results in a potentially advantageous increase in HDL cholesterol. Further study in larger randomized trials and other chronic kidney disease populations is indicated. PMID- 18385390 TI - Lessons for successful study enrollment from the Veterans Affairs/National Institutes of Health Acute Renal Failure Trial Network Study. AB - BACKGROUND AND OBJECTIVES: Design elements of clinical trials can introduce recruitment bias and reduce study efficiency. Trials involving the critically ill may be particularly prone to design-related inefficiencies. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Enrollment into the Veterans Affairs/National Institutes of Health Acute Renal Failure Trial Network Study was systematically monitored. Reasons for nonenrollment into this study comparing strategies of renal replacement therapy in critically ill patients with acute kidney injury were categorized as modifiable or nonmodifiable. RESULTS: 4339 patients were screened; 2744 fulfilled inclusion criteria. Of these, 1034 were ineligible by exclusion criteria. Of the remaining 1710 patients, 1124 (65.7%) enrolled. Impediments to informed consent excluded 21.4% of potentially eligible patients. Delayed identification of potential patients, physician refusal, and involvement in competing trials accounted for 4.4, 2.7, and 2.3% of exclusions. Comfort measures only status, chronic illness, chronic kidney disease, and obesity excluded 11.8, 7.8, 7.6, and 5.9% of potential patients. Modification of an enrollment window reduced the loss of patients from 6.6 to 2.3%. CONCLUSIONS: The Acute Renal Failure Trial Network Study's enrollment efficiency compared favorably with previous intensive care unit intervention trials and supports the representativeness of its enrolled population. Impediments to informed consent highlight the need for nontraditional acquisition methods. Restrictive enrollment windows may hamper recruitment but can be effectively modified. The low rate of physician refusal acknowledges clinical equipoise in the study design. Underlying comorbidities are important design considerations for future trials that involve the critically ill with acute kidney injury. PMID- 18385392 TI - Recruitment of live donors by candidates for kidney transplantation. AB - BACKGROUND: Little is known about efforts that renal transplant candidates make to recruit live donors. It was hypothesized that preference for live donor kidney transplantation and greater knowledge about live donor transplantation are associated with candidates' initiating conversations about donation with potential donors. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A cross sectional study of renal transplant candidates was performed at initial transplant evaluation. Candidates completed a questionnaire that specified whether they had initiated conversations about donation with any potential donors. The questionnaire also measured preference for live donor transplantation, knowledge about transplantation, concern about donor harm, willingness to ask for help in coping with kidney disease, and social support. RESULTS: Ninety-six candidates participated. Forty-nine (51%) reported initiating a conversation with at least one potential donor. In multivariable logistic regression, domains associated with initiating a conversation included: preference for live donor transplantation, willingness to ask for help, and female gender. Older age was associated with a lower odds of initiating a conversation. Knowledge, concern about donor harm, social support, and ethnicity were not associated with initiating a conversation with a donor. CONCLUSIONS: Attempts at donor recruitment by kidney transplant candidates are common. These findings suggest that interventions that influence preferences about transplantation and willingness to ask others for help are logical targets to enhance access to live donor transplantation. PMID- 18385393 TI - Variations in the risk for cerebrovascular events after kidney transplant compared with experience on the waiting list and after graft failure. AB - BACKGROUND AND OBJECTIVES: This study examined the risks, predictors, and mortality implications of cerebrovascular disease events after kidney transplantation in a national cohort. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This analysis used United States Renal Data System registry data to study retrospectively Medicare-insured kidney transplant candidates (n = 51,504), recipients (n = 29,614), and recipients with allograft failure (n = 2954) in 1995 through 2002. New-onset cerebrovascular disease events including ischemic stroke, hemorrhagic stroke, and transient ischemic attacks were ascertained from billing records, and participants were followed until Medicare-end or December 31, 2002. Multivariable survival analysis was used to compare cerebrovascular disease event incidence and risk profiles among the study samples. RESULTS: The cumulative, 3 yr incidence of de novo cerebrovascular disease events after transplantation was 6.8% and was lower than adjusted 3-yr estimates of 11.8% on the waiting list and 11.2% after graft loss. In time-dependent regression, transplantation predicted a 34% reduction in subsequent, overall cerebrovascular disease events risk compared with remaining on the waiting list, whereas risk for cerebrovascular disease events increased >150% after graft failure. Similar relationships with transplantation and graft loss were observed for each type of cerebrovascular disease event. Smoking was a potentially preventable correlate of posttransplantation cerebrovascular disease events. Women were not protected. All forms of cerebrovascular disease event diagnoses after transplantation predicted increased mortality. CONCLUSIONS: Along with known benefits for cardiac complications, transplantation with sustained graft function seems to reduce risk for vascular disease events involving the cerebral circulation. PMID- 18385394 TI - Renal biopsies in acute kidney injury: who are we missing? PMID- 18385395 TI - Leflunomide therapy in kidney transplantation: ready for prime time? PMID- 18385396 TI - How should nephrologists approach gadolinium-based contrast imaging in patients with kidney disease? PMID- 18385397 TI - Ultrastructural evidence of dermal gadolinium deposits in a patient with nephrogenic systemic fibrosis and end-stage renal disease. AB - BACKGROUND AND OBJECTIVES: The pathogenesis of acquired nephrogenic systemic fibrosis recently described for patients with renal insufficiency and a history of exposition to gadolinium-based magnetic resonance contrast agents is not completely understood. A role for circulating fibroblasts in the fibrosing tissue is hypothetical, and the mechanism of the assumed trigger function of gadolinium remains elusive. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: A skin lesion on a 76-yr-old man with symptoms of nephrogenic systemic fibrosis lasting 5 mo was studied at the ultrastructural level. After confirmation of he diagnosis by histopathologic methods, the presence and distribution of gadolinium, iron, calcium, and magnesium by energy filtering transmission electron microscopy was also examined. RESULTS: The performed electron spectroscopic imaging and electron energy loss spectroscopic analyses on deparaffinized samples revealed deposition of gadolinium in irregular small aggregates that adhered to cell profiles and collagen fibers of the connective tissue, forming a perivascular "gadolinium deposit zone" in the skin. Traces of iron signal were demonstrated in singular gadolinium-positive deposits, and iron presence was found in adjacent connective tissue. The ultrastructural cell analysis of the lesion showed among numerous poorly differentiated fibrocytes also higher differentiated cells with myofibroblastic characteristics, including bundles of intermediate filaments and attachment plaques in the cell periphery, indicating an ability of lesional fibroblasts to differentiate into myofibroblastic cells. CONCLUSIONS: These findings support the pivotal role of gadolinium chelates in the development of nephrogenic systemic fibrosis. PMID- 18385398 TI - Put the patient in the room, always. PMID- 18385399 TI - Medicine information needs of patients: the relationships between information needs, diagnosis and disease. AB - OBJECTIVE: To identify medicine information needs of patients and explore differences in information needs between different disease groups of patients. DESIGN: Semistructured interviews with general medical patients selected via convenience sampling. SETTING: Patients were recruited while inpatients during a hospital stay or as outpatients attending a specific clinic at the hospital. MAIN OUTCOME MEASURES: Patients' responses to standardised data-collection tools, including previously validated scale, the Extent of Information Desired scale (EID) to identify their information needs. RESULTS: Data from interviews with 1717 patients were included in the analysis. Each item on the EID scale was scored on a Likert scale (from 1 to 5). The internal consistency of the scale in this sample was good (coefficient alpha = 0.78). Scores to the EID scale correlated with age and socio-demographic variables. The extent of information desired positively correlated with socio-economic status (Pearson's r = 0.29, p<0.001). The extent of information desired negatively correlated with the patient's age (Pearson's r = -0.32, p<0.001), implying that medicine-information desires decreases with age. Subsequently, significant differences were found in the extent of information desired between disease categories, which remained significant when controlling for age (ANCOVA, F(6,1703) = 26.04, p<0.001, partial eta2 0.084 (ie, 8.4% "effect size"). Disease categories included: cardiovascular, gastrointestinal, respiratory, endocrine, diabetic, oncology. Patients with endocrine and diabetes diagnoses expressed high desires for information, whereas patients with cardiovascular and respiratory diagnoses expressed low desires for information. From these findings, both the disease and the age of patient are principal influences on desires for medicine information. CONCLUSIONS: These findings suggest that the diagnosis and disease have a significant bearing on patients' medicine-information desires and recommend that healthcare professionals view patients as individuals when providing information that meets their needs. It will be important for healthcare professionals to identify and understand that patients with different diseases have different desires for information about their disease and their drugs which may influence the way they take their medicines and subsequently the ways we manage their long-term disease. We need to determine if the EID scale is an efficient and effective way to identify patients' desires for drug information and a useful tool for practitioners to effectively target interventions in healthcare provision over time. PMID- 18385400 TI - How willing are patients to question healthcare staff on issues related to the quality and safety of their healthcare? An exploratory study. AB - BACKGROUND: One in 10 patients admitted to hospital will suffer an adverse event as a result of their medical treatment. A reduction in adverse events could happen if patients could be engaged successfully in monitoring their care. OBJECTIVES: This study explored: (1) surgical patients' willingness to question healthcare staff about their treatment; (2) differences between patients' willingness to ask factual vs. challenging questions related to the quality and safety of their healthcare; (3) patient demographic characteristics that could affect patients' willingness to ask questions; and (4) the impact of doctors' instructions on patients' willingness to ask questions. DESIGN: Cross-sectional study using the Patient Willingness to Ask Safety Questions Survey (PWASQS). The PWASQS questions were devised in accordance with current patient safety initiatives aimed at encouraging patients to ask healthcare staff specific safety related questions about their healthcare. The PWASQS includes factual questions (eg, "when can I return to my normal activities?") and challenging questions (eg, "have you washed your hands?"), and examines the impact of doctors' instructions on patients' willingness to ask challenging questions (eg, if instructed to by a doctor would you be willing to ask: "have you washed your hands?"). Data were analysed using non-parametric tests. SETTING: An inner-city London teaching hospital. PARTICIPANTS: A convenience sample of 80 patients who had undergone surgery. FINDINGS: Surgical patients were significantly more willing to ask: doctors factual versus challenging questions (z = 7.59, p<0.001); nurses factual versus challenging questions (z = 5.39, p<0.001); doctors versus nurses factual questions (z = 4.98, p<0.001); and, nurses versus doctors challenging questions (z = 4.40, p<0.001). Doctor's instructions to the patient increased patient willingness to challenge doctors (z = 6.56, p<0.001) and nurses (z = 6.15, p<0.001).Women, educated patients, and patients in employment, were more willing to ask questions (p<0.05). CONCLUSION: Surgical patients, particularly those who are men, less educated or unemployed are less willing to challenge healthcare staff regarding their care than to ask healthcare staff factual questions. Patient involvement strategies which take into account patient characteristics need to be developed for patients and staff in order to encourage patient involvement in this much neglected area. PMID- 18385401 TI - Preferences of acutely ill patients for participation in medical decision-making. AB - OBJECTIVES: To determine patient preferences for information and for participation in decision-making, and the determinants of these preferences in patients recently admitted to an acute hospital. DESIGN: Prospective questionnaire-based study. SETTING: Medical wards of an acute teaching hospital. PARTICIPANTS: One hundred and fifty-two consecutive acute medical inpatients, median age 74 years. MEASUREMENTS: Standardised assessment included abbreviated mental test and subjective measure of severity of illness. Patients' desire for information was assessed using a 5-point Likert scale, and their desire for a role in medical decision-making using the Degner Control of Preferences Scale. RESULTS: Of the 152 patients, 93 (61%) favoured a passive approach to decision making (either "leave all decisions to the doctor" or "doctor makes final decision but seriously considers my opinion." In contrast, 101 (66%) patients sought "very extensive" or "a lot" of information about their condition. No significant effects of age, sex, socio-economic group or severity of acute illness on desire for information or the Degner scale result were found. There was no agreement between patients' preferences on the Degner scale and their doctors' predictions of those preferences. CONCLUSIONS: Acute medical inpatients want to receive a lot of information about their illness, but most prefer a relatively passive role in decision-making. The only way to determine individual patient preferences is to ask them; preferences cannot be predicted from clinical or sociodemographic data. PMID- 18385402 TI - "The Happy Migrant Effect": perceptions of negative experiences of healthcare by patients with little or no English: a qualitative study across seven language groups. AB - BACKGROUND: The subjective experiences of patients with little or no English who either attended the Emergency Department or were admitted to the wards of a tertiary referral hospital were explored. SETTING AND METHODS: The reports of experiences from 49 patients of a tertiary referral hospital were analysed using grounded theory methods applied to translated transcriptions from focus groups held with discharged patients in seven languages. RESULTS: Inability to communicate in English, poor patient and family involvement with staff, a lack of control or powerlessness, staff shortages, staff negligence or incompetence, and treatment delays were reported by some patients. Others tended to discount or minimise the significance of similar experiences, suggesting a construct, "The Happy Migrant Effect," in which there is reluctance to assert healthcare rights. Patients appear "happy" and satisfied, despite problems with their hospital care. Explanatory factors for the construct include extreme powerlessness related to being unable to communicate, a positive comparison of healthcare in the new country compared with the old, patriotism for the new country, cultural norms that proscribe acceptance, politeness or social desirability, self-denigration for not having learnt English and, for a few, a fear of reprisals if they spoke out in complaint. CONCLUSIONS: Some immigrant patients with poor language skills might not report serious problems with healthcare delivery. In all patients in this study where problems with healthcare were reported, the events were considered to be largely preventable by appropriate language facilitation, patient and family involvement, and provider respect and compassion. PMID- 18385403 TI - Use of a falls incident reporting system to improve care process documentation in nursing homes. AB - BACKGROUND: Falls are the most frequently reported adverse event among frail nursing home residents and are an important resident safety issue. Incident reporting systems have been successfully used to improve quality and safety in healthcare. The purpose of this study was to test the effect of a systematically guided menu-driven incident reporting system (MDIRS) on documentation of post fall evaluation processes in nursing homes. METHODS: Six for-profit nursing homes in southeastern USA participated in the study. Over a 4-month period, MDIRS was used in three nursing homes matched with another three nursing homes which continued using their existing narrative incident report to document falls. Trained geriatric nurse practitioner auditors used a data collection audit tool to collect medical record documentation of the processes of care for residents who fell. Multivariate analysis of covariance was used to compare the post-fall nursing care processes documented in the medical records. RESULTS: 207 medical records of resident who fell were examined. Over 75% of the sample triggered at high risk for falls by the minimum data set. An adequate neurological assessment was documented for only 18.4% of residents who had experienced a fall. Although two-thirds of the sample had a diagnosis of incontinence, less than 20% of the records had incontinence-related interventions in the nursing care plan. Overall, there was more complete documentation of the post-fall evaluation process in the medical records in nursing homes using the MDIRS than in nursing homes using standard narrative incident reports (p<0.001). CONCLUSION: Further improvements are necessary in reporting mechanisms to improve the post-fall assessment in nursing home residents. PMID- 18385404 TI - Causes of preventable drug-related hospital admissions: a qualitative study. AB - OBJECTIVE: To explore the causes of preventable drug-related admissions (PDRAs) to hospital. DESIGN: Qualitative case studies using semi-structured interviews and medical record review; data analysed using a framework derived from Reason's model of organisational accidents and cascade analysis. PARTICIPANTS: 62 participants, including 18 patients, 8 informal carers, 17 general practitioners, 12 community pharmacists, 3 practice nurses and 4 other members of healthcare staff, involved in events leading up to the patients' hospital admissions. SETTING: Nottingham, UK. RESULTS: PDRAs are associated with problems at multiple stages in the medication use process, including prescribing, dispensing, administration, monitoring and help seeking. The main causes of these problems are communication failures (between patients and healthcare professionals and different groups of healthcare professionals) and knowledge gaps (about drugs and patients' medical and medication histories). The causes of PDRAs are similar irrespective of whether the hospital admission is associated with a prescribing, monitoring or patient adherence problem. CONCLUSIONS: The causes of PDRAs are multifaceted and complex. Technical solutions to PDRAs will need to take account of this complexity and are unlikely to be sufficient on their own. Interventions targeting the human causes of PDRAs are also necessary--for example, improving methods of communication. PMID- 18385405 TI - Role of registered nurses in error prevention, discovery and correction. AB - BACKGROUND: Registered nurses have a vital role in discovering and correcting medical error. OBJECTIVE: To describe the type and frequency of errors detected by American critical care nurses, and to ascertain who made the errors discovered by study participants. METHODS: Daily logbooks were used to collect information about errors discovered by a random sample of 502 critical care nurses during a 28-day period. RESULTS: Although the majority of errors discovered and corrected by critical care nurses involved medications (163/367), procedural errors were common (n = 115). Charting and transcription errors were less frequently discovered. The errors discovered by participants were attributed to a wide variety of staff members including nurses, doctors, pharmacists, technicians and unit secretaries. CONCLUSIONS: Given the importance of nurses in maintaining patient safety, future studies should identify factors that enhance their effectiveness to prevent, intercept and correct healthcare errors. PMID- 18385406 TI - Resident uncertainty in clinical decision making and impact on patient care: a qualitative study. AB - BACKGROUND: Little is known regarding how internal medicine residents manage uncertainty during decision making and subsequent effects on patient care. The aims of this study were to describe types of uncertainty faced by residents, strategies employed to manage uncertainty and effects on patient care. METHODS: Using critical incident technique, residents were asked to recall important clinical decisions during a recent call night, with probes to identify decisions made during uncertainty. They were also asked to report who they approached for advice. Three authors independently coded transcripts using the constant comparative method. RESULTS: The 42/50 (84%) interviewed residents reported 18 discrete critical incidents. Six categories emerged and mapped to the domains of the Beresford Model of Clinical Uncertainty: technical uncertainty (procedural skills, knowledge of indications); conceptual uncertainty (care transitions, diagnostic decision making and management conflict) and personal uncertainty (goals of care). In managing uncertainty, residents report a "hierarchy of assistance", using colleagues and literature for initial management, followed by senior residents, specialty fellows and, finally, the attending physician. Barriers to seeking the attending physician's input included the existence of a defined hierarchy for assistance and fears of losing autonomy, revealing knowledge gaps, and "being a bother". For 12 of the 18 cases reported, patient care was compromised: delay in procedure or escalation of care (n = 8); procedural complications (n = 2); and cardiac arrest (n = 2). CONCLUSION: Resident uncertainty results in delays of indicated care and, in some cases, patient harm. Despite the presence of a supervisory figure, residents adhere to a hierarchy when seeking advice in clinical matters. PMID- 18385407 TI - Benefits and losses: a qualitative study exploring healthcare staff perceptions of teamworking. AB - OBJECTIVES: To examine staff perceptions of teamworking practice in the field of stroke care. DESIGN: Qualitative interview study. SETTING: Three teams providing care to patients with stroke across a typical care pathway of acute hospital ward, specialist stroke unit, and community rehabilitation. PARTICIPANTS: 37 staff members from a range of professions. MAIN OUTCOME MEASURES: Healthcare staff perceptions of teamworking. RESULTS: Through detailed coding and analysis of the transcripts, five perceptions regarding the impact of teamworking on staff and patients were identified. These were: (1) mutual staff support, (2) knowledge and skills sharing, (3) timely intervention/discharge, (4) reduced individual decision-making and responsibility and (5) impact on patient contact time. CONCLUSIONS: Teamworking practice may be associated with a number of perceived benefits for staff and patient care; however, the potential for losses resulting from reduced patient contact time and ill-defined responsibility needs further investigation. PMID- 18385408 TI - An analysis of decision letters by research ethics committees: the ethics/scientific quality boundary examined. AB - OBJECTIVES: The performance of NHS research ethics committees (RECs) is of growing interest. It has been proposed that they confine themselves to "ethical" issues only and not concern themselves with the quality of the science. This study aimed to identify current practices of RECs in relation to scientific issues in research ethics applications. METHODS: Letters written by UK RECs expressing provisional or unfavourable opinions in response to submitted research applications were sampled from the research ethics database held by the Central Office for Research Ethics Committees. Ethnographic content analysis (ECA) was used to develop a coding framework. QSR N6 software was used to facilitate coding. RESULTS: "Scientific issues" were raised in 104 (74%) of the 141 letters in our sample. The present data suggest that RECs frequently considered scientific issues and that judgments of these often informed their decisions about approval of applications. Current processes of peer review seemed insufficient to reassure RECs about the scientific quality of applications they were asked to review. CONCLUSIONS: This study provides evidence that scientific issues are frequently raised in letters to researchers and are often considered a quality problem by RECs. In the discussion, the authors reflect on how far issues of science can and should be distinguished from those of ethics and the policy implications. PMID- 18385409 TI - Plotting basic control charts: tutorial notes for healthcare practitioners. AB - There is considerable interest in the use of statistical process control (SPC) in healthcare. Although SPC is part of an overall philosophy of continual improvement, the implementation of SPC usually requires the production of control charts. However, as SPC is relatively new to healthcare practitioners and is not routinely featured in medical statistics texts/courses, there is a need to explain the issues involved in the selection and construction of control charts in practice. Following a brief overview of SPC in healthcare and preliminary issues, we use a tutorial-based approach to illustrate the selection and construction of four commonly used control charts (xmr-chart, p-chart, u-chart, c chart) using examples from healthcare. For each control chart, the raw data, the relevant formulae and their use and interpretation of the final SPC chart are provided together with a notes section highlighting important issues for the SPC practitioner. Some more advanced topics are also mentioned with suggestions for further reading. PMID- 18385410 TI - Decreasing paediatric prescribing errors in a district general hospital. AB - BACKGROUND: In paediatric inpatients, medication errors occur as frequently as 1 in 4.2 drug orders, with up to 80% of these being prescribing errors. CONTEXT: The children's unit of a district general hospital in West Yorkshire, UK. KEY MEASURES FOR IMPROVEMENT: Prescribing errors and preventable adverse drug events STRATEGIES FOR CHANGE: (1) The introduction of a junior doctor prescribing tutorial. (2) The introduction of a bedside prescribing guideline. EFFECTS OF CHANGE: The introduction of the junior doctor prescribing tutorial decreased the prescribing errors by 46%. The introduction of a bedside prescribing guideline did not decrease prescribing errors but may have been helpful to those doctors unable to attend a prescribing tutorial. LESSONS LEARNT: By investing time and providing appropriate written resources, we have been able to reduce our paediatric prescribing errors on the children's ward by almost half. PMID- 18385411 TI - Kaoru Ishikawa: from fishbones to world peace. PMID- 18385412 TI - Chronic liver disease--an increasing problem: a study of hospital admission and mortality rates in England, 1979-2005, with particular reference to alcoholic liver disease. AB - AIMS: To determine time trends in hospital admissions for chronic liver disease in England between 1989/1990 and 2002/2003, mortality rates in England and Wales between 1979 and 2005, and the influence of alcohol-related disease on these trends. METHODS: Hospital episode statistics for admissions in England were obtained from the Information Center for Health and Social Care and mortality data for England and Wales from the Office for National Statistics. RESULTS: Hospital admission rates for chronic liver disease increased by 71% in males and 43% in females over the study period. This increase was largely due to alcoholic liver disease, admission rates for which more than doubled between 1989/1990 and 2002/2003. While there was a smaller rise for chronic viral hepatitis B and C, admission rates declined for hepatitis A, autoimmune hepatitis, and primary biliary cirrhosis. Mortality rates for chronic liver disease more than doubled between 1979 and 2005. Two thirds of these deaths were attributable to alcohol related liver disease in 2005. The highest rate of alcoholic liver disease mortality was in the 45-64 age group, and the largest percentage increase between 1979 and 2005 occurred in the 25-34 age group. CONCLUSIONS: Hospital admissions and mortality in England from chronic liver disease are increasing. The underlying reasons are complex, but alcohol-induced liver disease makes a major contribution. There are clear social and health implications if the trend continues and addressing alcohol-related liver disease should be a public health priority. PMID- 18385414 TI - Toward the promise of renal replacement therapy. PMID- 18385413 TI - The pervading influence of alcoholic liver disease in hepatology. AB - Rising levels of alcohol consumption in the UK are leading to substantial increases in morbidity and mortality from liver disease. Drinking is starting at an earlier age with binging an increasing common pattern, and women are overtaking men in the consumption. Manifestations of liver damage range from fatty liver to end-stage cirrhosis, but it is the increasing number of cases presenting with an acute alcoholic hepatitis (AAH) that are the cause for greatest concern. Development of well-validated prognostic scoring systems (Maddrey Modified Discriminant Function, Glasgow Alcohol Score) makes it possible to select those patients with AAH who are most likely to respond to corticosteroids. The results of early pilot studies of a number of anti-TNF agents are encouraging and with infliximab, reduction in portal pressure has been demonstrated to be consequent on controlling inflammatory processes in the liver. For those deteriorating to the stage of liver failure, artificial liver support with MARS is of value in correcting major pathophysiological disturbances and as a bridge to liver transplantation, the results of which both for end-stage alcoholic cirrhosis and for AAH--of which there is limited experience, are excellent. Even as the stringent regulatory measures needed to control rising alcohol consumption are introduced by government, the burden of liver disease in the UK will remain high for years to come. PMID- 18385415 TI - Some assembly required: renal hypodysplasia and the problem with faulty parts. PMID- 18385416 TI - Perinatal nephron programming is not so sweet in maternal diabetes. PMID- 18385417 TI - Just add water. PMID- 18385419 TI - Staging of chronic kidney disease: time for a course correction. AB - Awareness of chronic kidney disease (CKD) has increased in part because of the definitions and treatment guidelines set out by Kidney Disease Outcomes Quality Initiative (KDOQI); however, the staging system set forth by these guidelines has led to several problems and unforeseen consequences. Stages 1 and 2 CKD are difficult to determine using the standard Modification of Diet in Renal Disease (MDRD) estimation of GFR, and their clinical significance in the absence of other risk factors is unclear. Just because microalbuminuria in people without diabetes is a cardiovascular risk factor does not make it kidney disease. Most patients who receive a diagnosis of stage 3 CKD (GFR between 30 and 59 ml/min) are elderly people, and the vast majority of these patients will die before they reach ESRD. The staging system needs to be modified to reflect the severity and complications of CKD. It is suggested that stages 1 and 2 be eliminated and stages 3, 4, and 5, be simply termed moderate impairment, severe impairment, and kidney failure, respectively. In addition, age should be a modifying factor, especially in moderate kidney impairment. These changes would allow identification and treatment of clinically relevant disease and avoidance of what can seem exaggerated prevalence estimates. PMID- 18385420 TI - Association of a CYP4A11 variant and blood pressure in black men. AB - CYP4A11 arachidonic acid monooxygenase oxidizes endogenous arachidonic acid to 20 hydroxyeicosatetraenoic acid, a renal vasoconstrictor and natriuretic. Cyp4a deficiency causes hypertension in male mice, and a loss-of-function variant (T8590C) of CYP4A11 is associated with hypertension in white individuals. Hypertension and hypertensive renal disease are more common among black than white individuals, but the relationship between genetic variation at CYP4A11 and hypertension in black individuals is not known. This study tested the hypothesis that the CYP4A11 T8590C polymorphism is associated with higher BP or clinical outcomes in 732 black Americans with hypertensive renal disease participating in the African American Study of Kidney Disease (AASK). Men with the 8590CC genotype had significantly higher systolic BP (CC 156.5 +/- 22.6 versus 148.4 +/- 24.3 mmHg in CT and TT combined; P = 0.04) and pulse pressure (P = 0.04) at baseline; this association was not observed among women. In addition, this genotype was associated with higher systolic and diastolic BP at 36-mo follow-up among those randomly assigned to the lower BP arm of the AASK. Among all participants (or men but not women) with proteinuria, the 8590CC genotype was associated with an increased cumulative incidence of ESRD or death, controlling for randomization and clinical characteristics. In summary, the CYP4A11 8590CC genotype is associated with increased BP in black men with hypertensive nephrosclerosis and is associated with adverse clinical outcomes in those with baseline proteinuria. These data support a role for renal monooxygenases and 20-hydroxyeicosatetraenoic acid in the regulation of BP and renal function in men. PMID- 18385421 TI - Maternal environment interacts with modifier genes to influence progression of nephrotic syndrome. AB - Mutations in the NPHS2 gene, which encodes podocin, are responsible for some cases of sporadic and familial autosomal recessive steroid-resistant nephrotic syndrome. Inter- and intrafamilial variability in the progression of renal disease among patients bearing NPHS2 mutations suggests a potential role for modifier genes. Using a mouse model in which the podocin gene is constitutively inactivated, we sought to identify genetic determinants of the development and progression of renal disease as a result of the nephrotic syndrome. We report that the evolution of renal disease as a result of nephrotic syndrome in Nphs2 null mice depends on genetic background. Furthermore, the maternal environment significantly interacts with genetic determinants to modify survival and progression of renal disease. Quantitative trait locus mapping suggested that these genetic determinants may be encoded for by genes on the distal end of chromosome 3, which are linked to proteinuria, and on the distal end of chromosome 7, which are linked to a composite trait of urea, creatinine, and potassium. These loci demonstrate epistatic interactions with other chromosomal regions, highlighting the complex genetics of renal disease progression. In summary, constitutive inactivation of podocin models the complex interactions between maternal and genetically determined factors on the progression of renal disease as a result of nephrotic syndrome in mice. PMID- 18385422 TI - Sirolimus is associated with new-onset diabetes in kidney transplant recipients. AB - New-onset diabetes (NOD) is associated with transplant failure. A few single center studies have suggested that sirolimus is associated with NOD, but this is not well established. With the use of data from the United States Renal Data System, this study evaluated the association between sirolimus use at the time of transplantation and NOD among 20,124 adult recipients of a first kidney transplant without diabetes. Compared with patients treated with cyclosporine and either mycophenolate mofetil orazathioprine, sirolimus-treated patients were at increased risk for NOD, whether it was used in combination with cyclosporine (adjusted hazard ratio [HR] 1.61; 95% confidence interval [CI] 1.36 to 1.90),tacrolimus (adjusted HR 1.66; 95% CI 1.42 to 1.93), or an antimetabolite (mycophenolate mofetil orazathioprine; adjusted HR 1.36; 95% CI 1.09 to 1.69). Similar results were obtained in a subgroup analysis that included the 16,861 patients who did not have their immunosuppressive regimen changed throughout the first posttransplantation year. In conclusion, sirolimus is independently associated with NOD. Given the negative impact of NOD on posttransplantation outcomes, these findings should be confirmed in prospective studies or in meta analyses of existing trials that involved sirolimus. PMID- 18385423 TI - The effect of altitude on dosing and response to erythropoietin in ESRD. AB - For poorly understood reasons, patients with end-stage renal disease (ESRD) differ substantially in their response to treatment with recombinant erythropoietin (EPO). Because hypoxia influences many of the biologic pathways involved in erythropoiesis, the altitude at which a patient lives may affect the dose-response relationship of EPO. In this retrospective cohort study, clinical data from 341,737 incident hemodialysis patients registered in the U.S. Renal Data System were combined with elevation data from the U.S. Geological Survey to address this question. Higher altitude was associated with smaller EPO doses and higher hematocrit levels. For example, compared with patients at sea level, patients living above 6000 ft received 19% less EPO (12.9 versus 15.9 thousand units/wk) but had hematocrit levels 1.1 points higher (35.7% versus 34.6%). These associations were found within subgroups defined by sex, race, age, calendar time, cause of ESRD, and dialysis center profit status, and persisted after adjustment for various potential confounding factors. Furthermore, resistance to EPO decreased with elevation. Our results suggest that ESRD patients living at high altitude either increase endogenous EPO production or respond better to endogenous and exogenous EPO. PMID- 18385424 TI - Increased risk of CKD among type 2 diabetics with nonalcoholic fatty liver disease. AB - It is unknown whether chronic kidney disease (CKD) is associated with nonalcoholic fatty liver disease among patients with type 2 diabetes. We followed 1760 outpatients with type 2 diabetes and normal or near-normal kidney function and without overt proteinuria for 6.5 yr for the occurrence of CKD (defined as overt proteinuria and/or estimated GFR <60 ml/min per 1.73 m(2)). During follow up, 547 participants developed incident CKD. Nonalcoholic fatty liver disease, diagnosed by liver ultrasound and exclusion of other common causes of chronic liver disease, was associated with a moderately increased risk for CKD (hazard ratio 1.69; 95% confidence interval 1.3 to 2.6; P < 0.001). Adjustments for gender, age, body mass index, waist circumference, BP, smoking, diabetes duration, glycosylated hemoglobin, lipids, baseline estimated GFR, microalbuminuria, and medications (hypoglycemic, lipid-lowering, antihypertensive, or antiplatelet drugs) did not appreciably attenuate this association (hazard ratio 1.49; 95% confidence interval 1.1 to 2.2; P < 0.01). In conclusion, our findings suggest that nonalcoholic fatty liver disease is associated with an increased incidence of CKD in individuals with type 2 diabetes, independent of numerous baseline confounding factors. PMID- 18385425 TI - LRRC50, a conserved ciliary protein implicated in polycystic kidney disease. AB - Cilia perform essential motile and sensory functions central to many developmental and physiological processes. Disruption of their structure or function can have profound phenotypic consequences, and has been linked to left right patterning and polycystic kidney disease. In a forward genetic screen for mutations affecting ciliary motility, we isolated zebrafish mutant hu255H. The mutation was found to disrupt an ortholog of the uncharacterized highly conserved human SDS22-like leucine-rich repeat(LRR)-containing protein LRRC50 (16q24.1) and Chlamydomonas Oda7p. Zebrafish lrrc50 is specifically expressed in all ciliated tissues. lrrc50(hu255H) mutants develop pronephric cysts with an increased proliferative index, severely reduced brush border, and disorganized pronephric cilia manifesting impaired localized fluid flow consistent with ciliary dysfunction. Electron microscopy analysis revealed ultrastructural irregularities of the dynein arms and misalignments of the outer-doublet microtubules on the ciliary axonemes, suggesting instability of the ciliary architecture in lrrc50(hu255H) mutants. TheSDS22-like leucine-rich repeats present in Lrrc50 are necessary for proper protein function, since injection of a deletion construct of the first LRR did not rescue the zebrafish mutant phenotype. Subcellular distribution of human LRRC50-EGFP in MDCK and HEK293T cells is diffusely cytoplasmic and concentrated at the mitotic spindle poles and cilium. LRRC50 RNAi knock-down in human proximal tubule HK-2 cells thoroughly recapitulated the zebrafish brush border and cilia phenotype, suggesting conservation of LRRC50 function between both species. In summary, we present the first genetic vertebrate model for lrrc50 function and propose LRRC50 to be a novel candidate gene for human cystic kidney disease, involved in regulation of microtubule-based cilia and actin-based brush border microvilli. PMID- 18385426 TI - Acute kidney injury leads to inflammation and functional changes in the brain. AB - Although neurologic sequelae of acute kidney injury (AKI) are well described, the pathogenesis of acute uremic encephalopathy is poorly understood. This study examined the short-term effect of ischemic AKI on inflammatory and functional changes of the brain in mice by inducing bilateral renal ischemia for 60 min and studying the brains 24 h later. Compared with sham mice, mice with AKI had increased neuronal pyknosis and microgliosis in the brain. AKI also led to increased levels of the proinflammatory chemokines keratinocyte-derived chemoattractant and G-CSF in the cerebral cortex and hippocampus and increased expression of glial fibrillary acidic protein in astrocytes in the cortex and corpus callosum. In addition, extravasation of Evans blue dye into the brain suggested that the blood-brain barrier was disrupted in mice with AKI. Because liver failure also leads to encephalopathy, ischemic liver injury was induced in mice with normal renal function; neuronal pyknosis and glial fibrillary acidic protein expression were not increased, suggesting differential effects on the brain depending on the organ injured. For evaluation of the effects of AKI on brain function, locomotor activity was studied using an open field test. Mice subjected to renal ischemia or bilateral nephrectomy had moderate to severe declines in locomotor activity compared with sham-operated mice. These data demonstrate that severe ischemic AKI induces inflammation and functional changes in the brain. Targeting these pathways could reduce morbidity and mortality in critically ill patients with severe AKI. PMID- 18385427 TI - Small-molecule CFTR inhibitors slow cyst growth in polycystic kidney disease. AB - Cyst expansion in polycystic kidney disease (PKD) involves progressive fluid accumulation, which is believed to require chloride transport by the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Herein is reported that small-molecule CFTR inhibitors of the thiazolidinone and glycine hydrazide classes slow cyst expansion in in vitro and in vivo models of PKD. More than 30 CFTR inhibitor analogs were screened in an MDCK cell model, and near-complete suppression of cyst growth was found by tetrazolo-CFTR(inh)-172, a tetrazolo derived thiazolidinone, and Ph-GlyH-101, a phenyl-derived glycine hydrazide, without an effect on cell proliferation. These compounds also inhibited cyst number and growth by >80% in an embryonic kidney cyst model involving 4-d organ culture of embryonic day 13.5 mouse kidneys in 8-Br-cAMP-containing medium. Subcutaneous delivery of tetrazolo-CFTR(inh)-172 and Ph-GlyH-101 to neonatal, kidney-specific PKD1 knockout mice produced stable, therapeutic inhibitor concentrations of >3 microM in urine and kidney tissue. Treatment of mice for up to 7 d remarkably slowed kidney enlargement and cyst expansion and preserved renal function. These results implicate CFTR in renal cyst growth and suggest that CFTR inhibitors may hold therapeutic potential to reduce cyst growth in PKD. PMID- 18385428 TI - Racial differences in mortality among those with CKD. AB - Compared with white individuals, black individuals have a significantly higher risk for death in the general population but seem to have a survival advantage in the ESRD population. Data on the relationship of race to survival in early stages of chronic kidney disease (CKD) are inconsistent. This study evaluated racial differences in mortality among the adult participants of the Third National Health and Nutrition Examination Survey, a population-based survey of community dwelling individuals. CKD was defined either by an estimated GFR < 60 ml/min per 1.73 m2 or by the presence of albuminuria, and this status was determined for 14,611 individuals, 2892 of whom were found to have CKD. Adjusting for age,gender, and race, risk for all-cause mortality among individuals with CKD was more than double that of individuals with normal renal function. In the subgroup with CKD, adjusting for age and gender,black individuals had a significantly higher risk for death, and this risk was modified by age;specifically, black individuals who were younger than 65 yr were 78% more likely to die than white individuals, whereas no significant differences in mortality were observed among individuals who were > or = 65 yr of age. Further adjustment for cardiovascular risk factors and CKD stage did not materially change the results, but the hazard ratios were significantly attenuated after adjustment for socioeconomic factors. In conclusion, these data demonstrate racial/ethnic disparities in mortality among individuals with CKD. This higher risk for death in early stages of CKD may explain the apparent survival advantage observed among black individuals who live long enough to reach stage 5 CKD. PMID- 18385430 TI - Attorney views on the use of private agencies for probation supervision and treatment. AB - The exponential rise in probation caseloads has necessitated that some jurisdictions contract with private probation agencies and community-based private treatment providers. Regulations that monitor private providers are ill defined or absent, leaving standards open to broad interpretation. Attorneys in one jurisdiction that used private providers were surveyed to measure their beliefs about private treatment providers for people sentenced to probation. The research found that attorneys supported private provider representatives being present in court to contact defendants and to provide evidence to strengthen probation violations. Private agency reputation and standardized criteria were important to establish qualifications in offering supervision and treatment services. PMID- 18385429 TI - Src inhibition ameliorates polycystic kidney disease. AB - Despite identification of the genes responsible for autosomal dominant polycystic kidney disease (PKD) and autosomal recessive PKD (ARPKD), the precise functions of their cystoprotein products remain unknown. Recent data suggested that multimeric cystoprotein complexes initiate aberrant signaling cascades in PKD, and common components of these signaling pathways may be therapeutic targets. This study identified c-Src (pp60(c-Src)) as one such common signaling intermediate and sought to determine whether Src activity plays a role in cyst formation. With the use of the nonorthologous BPK murine model and the orthologous PCK rat model of ARPKD, greater Src activity was found to correlate with disease progression. Inhibition of Src activity with the pharmacologic inhibitor SKI-606 resulted in amelioration of renal cyst formation and biliary ductal abnormalities in both models. Furthermore, the effects of Src inhibition in PCK kidneys suggest that the ErbB2 and B-Raf/MEK/ERK pathways are involved in Src-mediated signaling in ARPKD and that this occurs without reducing elevated cAMP. These data suggest that Src inhibition may provide therapeutic benefit in PKD. PMID- 18385431 TI - Perpetrators of sexual assault continuing to have sex with their victims following the initial assault: evidence for evolved reproductive strategies. AB - Among a sample of college students, roughly 30% of the women and 12% of the men reported having been the victim of a sexual assault sometime in their lives. Of the assault victims, approximately 23% of both sexes stated that they had sexual intercourse with their assaulters on at least one subsequent occasion. Female victims of a completed sexual assault were significantly more likely to continue being sexually active with their assailants than were female victims who managed to block the assault, while no such difference was found for male victims. This would imply that some men are using assaultive tactics to secure sex partners beyond a single sexual episode, thereby enhancing their potential reproductive success in evolutionary terms. Also, men who committed sexual assault reported having had more lifetime sex partners than did sexually experienced men with no sexual assault history. Overall, the idea that sexual assault is part of an evolved reproductive strategy is consistent with findings from this study. PMID- 18385432 TI - Breastfeeding rates in Iran: why is there such a large gap between Iran and other countries? PMID- 18385433 TI - Bacterial profiles of root caries in elderly patients. AB - Culture-based studies have shown that Streptococcus mutans and lactobacilli are associated with root caries (RC). The purpose of the present study was to assess the bacterial diversity of RC in elderly patients by use of culture-independent molecular techniques and to determine the associations of specific bacterial species or bacterial communities with healthy and carious roots. Plaque was collected from root surfaces of 10 control subjects with no RC and from 11 subjects with RC. The bacterial 16S rRNA genes from extracted DNA were PCR amplified, cloned, and sequenced to determine species identity. From a total of 3,544 clones, 245 predominant species or phylotypes were observed, representing eight bacterial phyla. The majority (54%) of the species detected have not yet been cultivated. Species of Selenomonas and Veillonella were common in all samples. The healthy microbiota included Fusobacterium nucleatum subsp. polymorphum, Leptotrichia spp., Selenomonas noxia, Streptococcus cristatus, and Kingella oralis. Lactobacilli were absent, S. mutans was present in one, and Actinomyces spp. were present in 50% of the controls. In contrast, the microbiota of the RC subjects was dominated by Actinomyces spp., lactobacilli, S. mutans, Enterococcus faecalis, Selenomonas sp. clone CS002, Atopobium and Olsenella spp., Prevotella multisaccharivorax, Pseudoramibacter alactolyticus, and Propionibacterium sp. strain FMA5. The bacterial profiles of RC showed considerable subject-to-subject variation, indicating that the microbial communities are more complex than previously presumed. The data suggest that putative etiological agents of RC include not only S. mutans, lactobacilli, and Actinomyces but also species of Atopobium, Olsenella, Pseudoramibacter, Propionibacterium, and Selenomonas. PMID- 18385434 TI - Catalase-negative Staphylococcus aureus strain with point mutations in the katA gene. AB - Methicillin-susceptible catalase-negative Staphylococcus aureus strain UCN61 was isolated from an arterial leg ulcer. The deduced sequence of the structural katA gene for the catalase was 99% identical to those of other S. aureus strains. Two mutations were identified in katA from S. aureus UCN61, including one leading to a substitution of key histidine 58 by a tyrosine. PMID- 18385435 TI - Novel human adenovirus causing nosocomial epidemic keratoconjunctivitis. AB - In 2000, we encountered cases of nosocomial infections with epidemic keratoconjunctivitis (EKC) at a university hospital in Kobe, in the western part of Japan. Two human adenovirus (HAdV) strains, Kobe-H and Kobe-S, were isolated from patients with nosocomial EKC infection. They were untypeable by existing neutralizing antisera; however, the isolate was neutralized with homologous antisera. We then encountered several cases of EKC due to nosocomial infections in eye clinics in different parts of Japan. A total of 80 HAdVs were isolated from patients with EKC at eight different hospitals. The partial hexon gene sequences of the isolates were determined and compared to those of the prototype strains of 51 serotypes. All isolates had identical partial hexon nucleotide sequences. Phylogenetic analysis classified these isolates into species of HAdV D. The isolates showed 93.9 to 96.7% nucleotide identity with HAdV-D prototype strains, while all 32 HAdV-D prototype strains ranged from 93.2 to 99.2% identity. The sequences of the loop 2 and fiber knob regions from the representative strain, Kobe-H, were dissimilar in all prototype strains of 51 serotypes. We believe that this virus is a novel serotype of HAdV that causes EKC. PMID- 18385436 TI - Is the Sequenced Bordetella pertussis strain Tohama I representative of the species? AB - Subtractive hybridization was carried out to identify differences between the sequenced genome of Bordetella pertussis Tohama I and those of two recently collected isolates. We identified genetic regions specific to recent isolates, old isolates, and isolates of B. parapertussis and B. bronchiseptica species. We conclude that Tohama I strain is not representative of the B. pertussis species. PMID- 18385437 TI - Detection of the Klebsiella pneumoniae carbapenemase type 2 Carbapenem hydrolyzing enzyme in clinical isolates of Citrobacter freundii and K. oxytoca carrying a common plasmid. AB - The Klebsiella pneumoniae carbapenemase (KPC) was detected in carbapenem resistant isolates of Citrobacter freundii and Klebsiella oxytoca recovered from different patients in a Michigan hospital. Restriction analysis and hybridization with a KPC-specific probe showed the bla(KPC-2) genes of these two genera of the family Enterobacteriaceae are carried on a common plasmid. PMID- 18385438 TI - Comparison of nasal swabs with nose blowing for community-based pneumococcal surveillance of healthy children. AB - The nasopharynx (NP) is the preferred site for detection of Streptococcus pneumoniae in young children, but NP sampling is not well tolerated. We compared nose blowing with paired nasal swabs. The sensitivity of nose blowing was 46% (95% confidence interval [CI] 38 to 56%), which increased to 94% (95% CI, 85 to 98%) for children with visible secretions. PMID- 18385439 TI - Identification of thermotolerant campylobacter species by fluorescence in situ hybridization. AB - Thermotolerant Campylobacter spp. (Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis) are leading causes of food-borne diarrhea in humans. In this study, the usefulness of fluorescence in situ hybridization (FISH) for the identification of Campylobacter isolates was investigated. A hierarchical FISH probe set that included six group-, genus-, and species-specific probes was developed and evaluated with 12 reference strains and 94 clinical isolates of Campylobacter, Arcobacter, and Helicobacter. FISH correctly identified all isolates to the genus level and detected all thermotolerant Campylobacter isolates. The assay showed high degrees of sensitivity for the identification of C. jejuni (90%), C. coli (97%), C. lari (81%), and C. upsaliensis (100%) to the species level. PMID- 18385440 TI - Evaluation of novel broad-range real-time PCR assay for rapid detection of human pathogenic fungi in various clinical specimens. AB - In the present study, a novel broad-range real-time PCR was developed for the rapid detection of human pathogenic fungi. The assay targets a part of the 28S large-subunit ribosomal RNA (rDNA) gene. We investigated its application for the most important human pathogenic fungal genera, including Aspergillus, Candida, Cryptococcus, Mucor, Penicillium, Pichia, Microsporum, Trichophyton, and Scopulariopsis. Species were identified in PCR-positive reactions by direct DNA sequencing. A noncompetitive internal control was applied to prevent false negative results due to PCR inhibition. The minimum detection limit for the PCR was determined to be one 28S rDNA copy per PCR, and the 95% detection limit was calculated to 15 copies per PCR. To assess the clinical applicability of the PCR method, intensive-care patients with artificial respiration and patients with infective endocarditis were investigated. For this purpose, 76 tracheal secretion samples and 70 heart valve tissues were analyzed in parallel by real-time PCR and cultivation. No discrepancies in results were observed between PCR analysis and cultivation methods. Furthermore, the application of the PCR method was investigated for other clinical specimens, including cervical swabs, nail and horny skin scrapings, and serum, blood, and urine samples. The combination of a broad-range real-time PCR and direct sequencing facilitates rapid screening for fungal infection in various clinical specimens. PMID- 18385441 TI - Evaluation of a rapid new stool antigen test for diagnosis of Helicobacter pylori infection in adult patients. AB - The evaluation of a new rapid stool antigen test showed different levels of sensitivity for final readings of test results at 20 min (59.1%) and 30 min (76.9%). Significant differences in performance were observed between the two sexes and the various age categories, with higher efficiency in male patients and young adults. Generally, this test is efficient and can be used to detect H. pylori infection in adults. However, further studies are required to confirm its accuracy. PMID- 18385442 TI - Oligonucleotide array-based identification of species in the Acinetobacter calcoaceticus-A. baumannii complex in isolates from blood cultures and antimicrobial susceptibility testing of the isolates. AB - Acinetobacter calcoaceticus, A. baumannii, Acinetobacter genomic species (gen. sp.) 3, and Acinetobacter gen. sp. 13TU, which are included in the A. calcoaceticus-A. baumannii complex, are difficult to distinguish by phenotypic methods. An array with six oligonucleotide probes based on the 16S-23S rRNA gene intergenic spacer (ITS) region was developed to differentiate species in the A. calcoaceticus-A. baumannii complex. Validation of the array with a reference collection of 52 strains of the A. calcoaceticus-A. baumannii complex and 137 strains of other species resulted in an identification sensitivity and specificity of 100%. By using the array, the species distribution of 291 isolates of the A. calcoaceticus-A. baumannii complex from patients with bacteremia were determined to be A. baumannii (221 strains [75.9%]), Acinetobacter gen. sp. 3 (67 strains [23.0%]), Acinetobacter gen. sp. 13TU (2 strains [0.7%]), and unidentified Acinetobacter sp. (1 strain [0.3%]). The identification accuracy of the array for 12 randomly selected isolates from patients with bacteremia was further confirmed by sequence analyses of the ITS region and the 16S rRNA gene. Antimicrobial susceptibility testing of the 291 isolates from patients with bacteremia revealed that A. baumannii strains were less susceptible to antimicrobial agents than Acinetobacter gen. sp. 3. All Acinetobacter gen. sp. 3 strains were susceptible to ampicillin-sulbactam, imipenem, and meropenem; but only 67.4%, 90%, and 86% of the A. baumannii strains were susceptible to ampicillin-sulbactam, imipenem, and meropenem, respectively. The observed significant variations in antimicrobial susceptibility among different species in the A. calcoaceticus-A. baumannii complex emphasize that the differentiation of species within the complex is relevant from a clinical-epidemiological point of view. PMID- 18385443 TI - Performance of HerpeSelect and Kalon assays in detection of antibodies to herpes simplex virus type 2. AB - The performances of commercial enzyme-linked immunosorbent assays (ELISAs) in detecting herpes simplex virus type 2 (HSV-2) antibodies have been inconsistent for African and human immunodeficiency virus (HIV)-positive populations. We compared the performances of the HerpeSelect and Kalon glycoprotein G2 ELISAs for patients with genital ulcer disease in Ghana and the Central African Republic. Sera from 434 women were tested with the HerpeSelect assay, and a subsample (n = 199) was tested by the Kalon assay. Ulcer swabs and cervicovaginal lavage samples were tested for HSV-2 DNA by PCR. HSV-2-seronegative women with detectable genital HSV-2 DNA were retested for HSV-2 antibodies 14 and 28 days later by the two ELISAs. A total of 346 (80%) women were positive by HerpeSelect at baseline, and 225 (54%) had detectable genital (lesional or cervicovaginal) HSV-2 DNA. Sixty-six (19%) HerpeSelect-positive samples had low-positive index values (1.1 to 3.5), and 58% of these samples had detectable genital HSV-2 DNA. Global agreement between the two serological assays was 86%. Concordance was high (99%) for sera that were negative by HerpeSelect or had high index values (>3.5). Defining infection detected by HSV-2 DNA PCR and/or Kalon assay as true infection, 71% of sera with low-positive index values were associated with true HSV-2 infection. Twenty-five women were identified as having nonprimary first episode genital HSV-2 infection. Rates of HSV-2 seroconversion at day 14 were 77% (10/13 patients) by HerpeSelect assay and 23% (3/13 patients) by Kalon assay, with four additional seroconversions detected by Kalon assay at day 28. HIV serostatus did not influence assay performance. Low index values obtained with the HerpeSelect assay may correspond to true HSV-2 infection, in particular to nonprimary first episodes of genital HSV-2 infection, and need to be interpreted in the context of clinical history. PMID- 18385444 TI - Pulmonary paragonimiasis diagnosed by fine-needle aspiration biopsy. AB - We report a case of paragonimiasis involving a 12-year-old Latin American boy. The diagnosis was made by fine-needle aspiration biopsy of a pulmonary nodule. Identification of the species by morphometric analysis of the eggs indicated that the infection was caused by Paragonimus mexicanus. PMID- 18385446 TI - Differential effects of prenatal testosterone timing and duration on phenotypic and behavioral masculinization and defeminization of female sheep. AB - The process of sexual differentiation leaves genetically female individuals at risk of being masculinized by exogenous androgens. Previous research with sheep indicates that exposure to excess testosterone from Gestational Day (GD) 30 to GD 90 of the 147-day gestation masculinizes and defeminizes behavior as well as genitalia. Lower doses and shorter durations produce animals with varying degrees of genital virilization and alterations of the hypothalamic-pituitary-gonadal axis, but to our knowledge, the effects on complex behavior and its prediction by the amount of external virilization have not been explored. Previous research in rodents has suggested that sexual differentiation of the central nervous system and the external genitalia can be dissociated. Therefore, we hypothesized that the extent of virilization of external genitalia would not be predictive of the lack of female-typical, or the presence of male-typical, mating behavior. To test this hypothesis, we compared control females, females exposed to exogenous testosterone from GD 30 to GD 90 (T60 females) that have virilized genitalia, and females exposed to testosterone from GD 60 to GD 90 (T30 females) that have female-typical genitalia. Both natural behavioral estrus in the flock and hormonally controlled behavioral tests were used to explore reproductive behavior. The T60 and T30 females exhibited more masculinized reproductive behavior than the controls; however, the T30 females also exhibited feminine behavior. Neither testosterone-treated group was receptive or was mounted at rates comparable to those of controls. These data illustrate that variation in the timing or duration of exposure to prenatal testosterone during a critical period for masculinization can have variable effects on defeminization and that the effects of testosterone on genitalia are not entirely predictive of behavior. PMID- 18385447 TI - Characterization and regulation of monocarboxylate cotransporters Slc16a7 and Slc16a3 in preimplantation mouse embryos. AB - Concurrent with compaction, preimplantation mouse embryos switch from the high pyruvate consumption that prevailed during cleavage stages to glucose consumption against a constant background of pyruvate uptake. However, zygotes exposed to and subsequently deprived of glucose can form blastocysts by increasing pyruvate uptake. This metabolic switch requires cleavage-stage exposure to glucose and is one aspect of metabolic differentiation that normally occurs in vivo. Monocarboxylates, such as pyruvate and lactate, are transported across membranes via the SLC16 family of H(+)-monocarboxylate cotransporter (MCT) proteins. Thus, the increase in pyruvate uptake in embryos developing without glucose must involve changes in activity and localization of MCT. In mouse embryos, continued expression of Slc16a1 (MCT1) requires glucose supply. Messenger RNA for Slc17a7 (MCT2) and Slc16a3 (MCT4) has been detected in mouse preimplantation embryos; however, protein function, localization, and regulation of expression at the basis of these net pyruvate uptake changes remain unclear. The expression and localization of SLC16A7 and SLC16A3 have therefore been examined to clarify their respective roles in embryos derived from the reproductive tract and cultured under varied conditions. SLC16A3 appears localized to the plasma membrane until the morula stage and also maintains a nuclear distribution throughout preimplantation development. However, continued Slc16a3 mRNA expression is dependent on prior exposure to glucose. SLC16A7 localizes to apical cortical regions with punctate, vesicular expression throughout blastomeres, partially colocalizing in peroxisomes with peroxisomal catalase (CAT). In contrast to SLC16A3 and SLC16A1, SLC16A7 and CAT demonstrate upregulation in the absence of glucose. These striking differences between the two isoforms in expression localization and regulation suggest unique roles for each in monocarboxylate transport and pH regulation during preimplantation development, and implicate peroxisomal SLC16A7 as an important redox regulator in the absence of glucose. PMID- 18385448 TI - A catenary model to study transport and conjugation of baicalein, a bioactive flavonoid, in the Caco-2 cell monolayer: demonstration of substrate inhibition. AB - The transport and metabolism of baicalein (Ba) was studied in vitro and in Caco-2 cells. Protein binding of Ba with Caco-2 lysate showed that Ba was bound to two classes of sites: a higher affinity, lower capacity site (K(A1) = 27.6 +/- 4.7 microM(-1), n(1) = 10.6 +/- 0.6 nmol/mg) and lower affinity, higher capacity site (K(A2) = 0.015 +/- 0.0013 microM(-1), n(2) = 413 +/- 21 nmol/mg). Incubation studies of Ba with Caco-2 lysate showed substrate inhibition of both glucuronidation and sulfation, with K(m) values of 0.14 +/- 0.034 and 0.015 +/- 0.0053 microM, and K(I) values of 6.75 +/- 1.70 and 0.37 +/- 0.16 microM, respectively. In the Caco-2 monolayer, Ba (8-47 microM) displayed good apparent permeabilities (P(app)) across the membrane; P(app) was found to be increased with elevated loading concentration in both the absorptive and secretory directions. However, the efflux ratio was less than unity, negating the involvement of apical efflux transporters. The concentration ratios of Ba sulfate (BS) and glucuronide (BG) decreased with increased loading Ba concentration, suggesting that BS and BG are apically excreted via transporters, likely breast cancer resistance protein and multidrug resistance-associated protein 2, respectively. Data fit to the catenary model, composed of basolateral, cellular, and apical compartments, showed a low cellular unbound fraction (0.0019 +/- 0.00018), a high passive diffusion clearance (0.012 +/- 0.00029 ml/min/mg), and substrate inhibition, with sulfation being more readily saturated and inhibited than glucuronidation, as evidenced by smaller K(m) value (0.35 +/- 0.078 versus 1.95 +/- 0.57 microM) and K(I) value (0.58 +/- 0.20 versus 7.90 +/- 1.10 microM); these patterns paralleled those observed in the lysate incubation studies. The results showed that the catenary model aptly predicts substrate inhibition kinetics and offers significant and mechanistic insight into the transport and atypical metabolism of drugs in the Caco-2 monolayer. PMID- 18385445 TI - Endocrine antecedents of polycystic ovary syndrome in fetal and infant prenatally androgenized female rhesus monkeys. AB - Experimentally induced fetal androgen excess induces polycystic ovary syndrome like traits in adult female rhesus monkeys (Macaca mulatta). Developmental changes leading to this endocrinopathy are not known. We therefore studied 15 time-mated, gravid female rhesus monkeys with known female fetuses. Nine dams received daily s.c. injections of 15 mg of testosterone propionate (TP), and six received injections of oil vehicle (control) from 40 through 80 days of gestation (term, 165 days; range, +/-10 days). All fetuses were delivered by cesarean section using established methods at term. Ultrasound-guided fetal blood sample collection and peripheral venous sample collection of dams and subsequent infants enabled determination of circulating levels of steroid hormones, LH and FSH. The TP injections elevated serum testosterone and androstenedione levels in the dams and prenatally androgenized (PA) fetuses. After cessation of TP injections, testosterone levels returned to values within the reference range for animals in these age groups, whereas serum androstenedione levels in PA infants were elevated. The TP injections did not increase estrogen levels in the dams or the PA fetuses or infants, yet conjugated estrogen levels were elevated in the TP injected dams. Serum levels of LH and FSH were elevated in late-gestation PA fetuses, and LH levels were elevated in PA infants. These studies suggest that experimentally induced fetal androgen excess increases gonadotropin secretion in PA female fetuses and infants and elevates endogenous androgen levels in PA infants. Thus, in this nonhuman primate model, differential programming of the fetal hypothalamo-pituitary unit with concomitant hyperandrogenism provides evidence to suggest developmental origins of LH and androgen excess in adulthood. PMID- 18385449 TI - Therapeutic potential of 1-methylnicotinamide against acute gastric lesions induced by stress: role of endogenous prostacyclin and sensory nerves. AB - 1-Methylnicotinamide (MNA) is one of the major derivatives of nicotinamide, which was recently shown to exhibit antithrombotic and antiinflammatory actions. However, it is not yet known whether MNA affects gastric mucosal defense. The effects of exogenous MNA were studied on gastric secretion and gastric lesions induced in rats by 3.5 h of water immersion and water restraint stress (WRS) or in rats administered 75% ethanol. MNA [6.25-100 mg/kg intragastrically (i.g.)] led to a dose-dependent rise in the plasma MNA level, inhibited gastric acid secretion, and attenuated these gastric lesions induced by WRS or ethanol. The gastroprotective effect of MNA was accompanied by an increase in the gastric mucosal blood flow and plasma calcitonin gene-related peptide (CGRP) levels, the preservation of prostacyclin (PGI(2)) generation (measured as 6-keto-PGF1alpha), and an overexpression of mRNAs for cyclooxygenase (COX)-2 and CGRP in the gastric mucosa. R-3-(4-Fluoro-phenyl)-2-[5-(4-fluoro-phenyl)-benzofuran-2 ylmethoxycarbonylamino]-propionic acid (RO 324479), which is the selective antagonist of IP/PGI(2) receptors, reversed the effects of MNA on gastric lesions and GBF. MNA-induced gastroprotection was attenuated by suppression of COX-1 [5 (4-chlorophenyl)-1-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazole; SC-560] and COX-2 [4-(4-methylsulfonylphenyl)-3-phenyl-5H-furan-2-one; rofecoxib] activity, capsaicin denervation, and by the pretreatment with CGRP(8-37) or capsazepine. Addition of exogenous PGI(2) or CGRP restored the MNA-induced gastroprotection in rats treated with COX-1 and COX-2 inhibitors or in those with capsaicin denervation. WRS enhanced MDA content while decreasing superoxide dismutase (SOD) activity in the gastric mucosa, but pretreatment with MNA reversed these changes. MNA exerts potent gastroprotection against WRS damage via mechanisms involving cooperative action of PGI(2) and CGRP in preservation of microvascular flow, antioxidizing enzyme SOD activity, and reduction in lipid peroxidation. PMID- 18385450 TI - ALK- anaplastic large-cell lymphoma is clinically and immunophenotypically different from both ALK+ ALCL and peripheral T-cell lymphoma, not otherwise specified: report from the International Peripheral T-Cell Lymphoma Project. AB - The International Peripheral T-Cell Lymphoma Project is a collaborative effort designed to gain better understanding of peripheral T-cell and natural killer (NK)/T-cell lymphomas (PTCLs). A total of 22 institutions in North America, Europe, and Asia submitted clinical and pathologic information on PTCLs diagnosed and treated at their respective centers. Of the 1314 eligible patients, 181 had anaplastic large-cell lymphoma (ALCL; 13.8%) on consensus review: One hundred fifty-nine had systemic ALCL (12.1%) and 22 had primary cutaneous ALCL (1.7%). Patients with anaplastic lymphoma kinase-positive (ALK(+)) ALCL had a superior outcome compared with those with ALK(-) ALCL (5-year failure-free survival [FFS], 60% vs 36%; P = .015; 5-year overall survival [OS], 70% vs 49%; P = .016). However, contrary to prior reports, the 5-year FFS (36% vs 20%; P = .012) and OS (49% vs 32%; P = .032) were superior for ALK(-) ALCL compared with PTCL, not otherwise specified (PTCL-NOS). Patients with primary cutaneous ALCL had a very favorable 5-year OS (90%), but with a propensity to relapse (5-year FFS, 55%). In summary, ALK(-) ALCL should continue to be separated from both ALK(+) ALCL and PTCL-NOS. Although the prognosis of ALK(-) ALCL appears to be better than that for PTCL-NOS, it is still unsatisfactory and better therapies are needed. Primary cutaneous ALCL is associated with an indolent course. PMID- 18385451 TI - The reactivity of Bw4+ HLA-B and HLA-A alleles with KIR3DL1: implications for patient and donor suitability for haploidentical stem cell transplantations. AB - Natural killer (NK)-cell alloreactivity can be exploited in haploidentical hematopoietic stem cell transplantation (HSCT). NK cells from donors whose HLA type includes Bw4, a public epitope present on a subset of HLA-B alleles, can be alloreactive toward recipients whose cells lack Bw4. Serologically detectable epitopes related to Bw4 also exist on a subset of HLA-A alleles, but the interaction of these alleles with KIR3DL1 is controversial. We therefore undertook a systematic analysis of the ability of most common HLA-B alleles and HLA-A alleles with Bw4 serologic reactivity to protect target cells from lysis by KIR3DL1-dependent NK cells. All Bw4(-) HLA-B alleles failed to protect target cells from lysis. All Bw4(+) HLA-B alleles with the exception of HLA-B*1301 and B*1302 protected targets from lysis. HLA-A*2402 and HLA-A*3201 unequivocally protected target cells from lysis, whereas HLA-A*2501 and HLA-A*2301 provided only weak protection from lysis. KIR3DL1-dependent alloreactive NK clones were identified in donors with HLA-A*2402 but not in donors with HLA-B*1301 or B*1302. These findings clarify the HLA types that donors and recipients need in haploidentical HSCT and other NK allotherapies in order to benefit from NK alloreactivity. PMID- 18385452 TI - Elevated protein tyrosine phosphatase activity provokes Eph/ephrin-facilitated adhesion of pre-B leukemia cells. AB - Signaling by Eph receptors and cell-surface ephrin ligands modulates adhesive cell properties and thereby coordinates cell movement and positioning in normal and oncogenic development. While cell contact-dependent Eph activation frequently leads to cell-cell repulsion, also the diametrically opposite response, cell-cell adhesion, is a probable outcome. However, the molecular principles regulating such disparate functions have remained controversial. We have examined cell biologic mechanisms underlying this switch by analyzing ephrin-A5-induced cell morphologic changes of EphA3-positive LK63 pre-B acute lymphoblastic leukemia cells. Their exposure to ephrin-A5 surfaces leads to a rapid conversion from a suspended/nonpolarized to an adherent/polarized cell type, a transition that relies on EphA3 functions operating in the absence of Eph-kinase signaling. Cell morphology change and adhesion of LK63 cells are effectively attenuated by endogenous protein tyrosine phosphatase (PTP) activity, whereby PTP inhibition and productive EphA3-phosphotyrosine signaling reverse the phenotype to nonadherent cells with a condensed cytoskeleton. Our findings suggest that Eph associated PTP activities not only control receptor phosphorylation levels, but as a result switch the response to ephrin contact from repulsion to adhesion, which may play a role in the pathology of hematopoietic tumors. PMID- 18385457 TI - Use of the Brucella melitensis native hapten to diagnose brucellosis in goats by a rapid, simple, and specific fluorescence polarization assay. AB - The performance of the fluorescence polarization assay (FPA) using the recently described Brucella melitensis native hapten and the Brucella abortus O polysaccharide tracer was evaluated and compared with those of The World Organization for Animal Health tests related to indirect and competitive enzyme linked immunosorbent assays as classification variables for goat sera obtained from a high-prevalence area where vaccination was performed; test series were also evaluated to increase the final specificity of the tests. Our results showed that the respective relative sensitivity and specificity were 99.7% and 32.5% for the rose Bengal test with a 3% cell concentration (RBT3), 92.8% and 68.8% for the rose Bengal test with 8% cell concentration (RBT8), 98.4% and 84.9% for the Canadian complement fixation test (CFT), 83.7% and 65.5% for the Mexican CFT, 98.4% and 81.0% for the buffered plate agglutination test (BPAT), and 78.1% and 89.3% for the fluorescence polarization assay (FPA). The use of the FPA as the secondary test significantly increased the final specificities of test combinations; the screening tests BPAT, RBT3, and RBT8 plus FPA resulted in 90%, 91.2%, and 91.3% final specificities, respectively, whereas for the combinations RBT3 plus Mexican CFT, RBT8 plus Mexican CFT, and BPAT plus Canadian CFT, the specificities were 65.5%, 63.2%, and 91.7%, respectively. The results suggested that the FPA may be routinely applied as an adaptable screening test for diagnosis of goat brucellosis, since its cutoff can be adjusted to improve its sensitivity or specificity, it is a rapid and simple test, it can be the test of choice when specificity is relevant or when an alternative confirmatory test is not available, and it is not affected by vaccination, thus reducing the number of goats wrongly slaughtered due to misdiagnosis. PMID- 18385458 TI - Distribution of dipeptidyl peptidase IV in patients with chronic tonsillitis. AB - In the pathogeneses of recurrent tonsillitis (RT) and tonsillar hypertrophy (TH), different immunological mechanisms are involved. Dipeptidyl peptidase IV (DPP IV) and aminopeptidase N (APN) participate in the regulation of the immune response during inflammation. In this study, the localization of DPP IV and the enzymatic activities of DPP IV and APN in 32 patients, 13 with RT and 19 with TH, who underwent tonsillectomy were investigated. The localization of DPP IV in tonsils was studied using histochemical and immunohistochemical methods. The enzymatic activities of DPP IV and APN in tonsillar lymphocytes and the patients' sera were determined kinetically at 37 degrees C using Gly-Pro-p-nitroanilide (for DPP IV) and Ala-p-nitroanilide (for APN) as chromogenic substrates. In samples from both RT and TH patients, DPP IV was found to localize mainly in extrafollicular areas of tonsillar tissue in a pattern corresponding to the T-cell distribution. Significantly higher (P < 0.001) levels of DPP IV and APN activities in sera from patients with TH than in sera from patients with RT were found. A correlation of DPP IV activities in sera and tonsillar lymphocytes from patients with TH was also found (r = 0.518; P < 0.05). Moreover, the results show that DPP IV and APN activities in sera decreased significantly with age. Tonsillar lymphocytes demonstrated a wide range of DPP IV and APN activities, without significant differences between the investigated groups. The results of this study show that the localization of DPP IV does not depend on the type of tonsillitis, whereas the variety in levels of DPP IV and APN activities in sera of patients with TH and RT suggests different patterns of participation of antigen-stimulated tonsils in the immune system. PMID- 18385459 TI - Expression of a functional single-chain variable-fragment antibody against complement receptor 1 in Streptococcus gordonii. AB - Streptococcus gordonii, an oral commensal organism, is a candidate vector for oral-vaccine development. Previous studies have shown that recombinant S. gordonii expressing heterologous antigens was weakly immunogenic when delivered intranasally. In this study, antigen was specifically targeted to antigen presenting cells (APC) in order to potentiate antigen-APC interactions and increase the humoral immune response to the antigen. To achieve this goal, a single-chain variable-fragment (scFv) antibody against complement receptor 1 (CR1) was constructed. Anti-CR1 scFv purified from Escherichia coli was able to bind to mouse mixed lymphocytes and bone marrow-derived dendritic cells. The in vivo function of the anti-CR1 scFv protein was assessed by immunizing mice intranasally with soluble scFv and determining the immune response against the hemagglutinin (HA) peptide located on the carboxy terminus of the scFv. The serum anti-HA immunoglobulin G (IgG) immune response was dose dependent; as little as 100 ng of anti-CR1 scFv induced a significant IgG immune response, while such a response was minimal when the animals were given an unrelated scFv. The anti-CR1 scFv was expressed in S. gordonii as a secreted protein, which was functional, as it bound to dendritic cells. Mice orally colonized by the anti-CR1-secreting S. gordonii produced an anti-HA IgG immune response, indicating that such an approach can be used to increase the immune response to antigens produced by this bacterium. PMID- 18385460 TI - Serological survey for foot-and-mouth disease virus in wildlife in eastern Africa and estimation of test parameters of a nonstructural protein enzyme-linked immunosorbent assay for buffalo. AB - In this study we estimate the seroprevalence of foot-and-mouth disease virus (FMDV) in wildlife from eastern and central Africa. Sera were sourced from between 1994 and 2002 from a rinderpest surveillance program. Our study compared a nonstructural protein enzyme-linked immunosorbent assay (Cedi test) with a virus neutralization test. The study shows that there is only a low seroprevalence of FMDV in sampled nonbuffalo species. The seroprevalence in the Cape buffalo was high for SAT2, lower for SAT1, and lowest for SAT3. As the SAT2 serotype was most prevalent, the Cedi test largely reflected the occurrence of SAT2-positive animals. The results also suggest that SAT2 became dominant around 1998, with a large increase in seroprevalence. The sensitivity and specificity of the Cedi test were estimated by comparison to the combined virus neutralization test results from all three SAT tests. A Bayesian implementation of the Hui Walter latent class model was used to estimate the test parameters. The model permits estimation in the absence of a gold standard test. The final model, using noninformative priors and assuming conditional independence of test performance, estimated Cedi test sensitivity at 87.7% and specificity at 87.3%. These estimates are similar to those for domestic bovines; they suggest that the Cedi test is a useful tool for screening buffalo for infection with the various serotypes of FMDV. PMID- 18385461 TI - Neonatal stem cells exhibit specific characteristics in function, proliferation, and cellular signaling that distinguish them from their adult counterparts. AB - Stem cells may be a novel treatment modality for organ ischemia, possibly through beneficial paracrine mechanisms. Stem cells from older hosts have been shown to exhibit decreased function during stress. We therefore hypothesized that 1) neonatal bone marrow mesenchymal stem cells (nBMSCs) would produce different levels of IL-6, VEGF, and IGF-1 compared with adults (aBMSCs) when stimulated with TNF or LPS; 2) differences in cytokines would be due to distinct cellular characteristics, such as proliferation or pluripotent potential; and 3) differences in cytokines would be associated with differences in p38 MAPK and ERK signaling within nBMSCs. BMSCs were isolated from adult and neonatal mice. Cells were exposed to TNF or LPS with or without p38 or ERK inhibition. Growth factors were measured via ELISA, proliferation via daily cell counts, cell surface markers via flow cytometry, and pluripotent potential via alkaline phosphatase activity. nBMSCs produced lower levels of IL-6 and VEGF, but higher levels of IGF 1 under basal conditions, as well as after stimulation with TNF, but not LPS. Neonatal and adult BMSCs had similar pluripotent potentials and cell surface markers, but nBMSCs proliferated faster. Furthermore, p38 and ERK appeared to play a more substantial role in nBMSC cytokine and growth factor production. Neonatal stem cells may aid in decreasing the local inflammatory response during ischemia, and could possibly be expanded more rapidly than adult cells prior to therapeutic use. PMID- 18385462 TI - The bone-muscle ratio of fetal lambs is affected more by maternal nutrition during pregnancy than by maternal size. AB - Bone formation and loss are related to the strain imposed on bone by muscle forces. Bone mineral content (BMC) and lean mass (LM) of fetal lambs was determined at day 140 of pregnancy in 8 groups of ewes, which were of either large or small body size, on either high (ad libitum) or maintenance pasture intake from day 21 of pregnancy, or carrying either singletons or twins. BMC and LM (using DXA scanning) of fetal hindquarters/spine were corrected to leg length. BMC and LM were less in twin than singleton groups (P < 0.001). Large ewes on high intake produced single fetuses with a (group mean) BMC/LM ratio that was higher (P < 0.002) than that in fetuses of large ewes with singletons on maintenance intake or twins on either high or maintenance intakes, the ratios of which were not different. In single fetuses from small ewes on high intake, the BMC/LM ratio was higher than those from small ewes with singletons on maintenance intake or twins on either high or maintenance intakes, the ratios of which were not different. The ratio was not different in singleton fetuses of ewes on high intake, whether they were large or small. Different fetal environments resulted in a given amount of muscle being associated with a higher or lower bone mass. Dietary intake during pregnancy was more important than maternal size in affecting the ratio. We conclude that intrauterine environmental factors may be important in determining bone mass postnatally, and possibly later in life. PMID- 18385463 TI - Hepatic insulin resistance induced by prenatal alcohol exposure is associated with reduced PTEN and TRB3 acetylation in adult rat offspring. AB - Prenatal alcohol exposure (EtOH) results in insulin resistance in rats of both sexes with increased expression of hepatic gluconeogenic genes and glucose production. To investigate whether hepatic insulin signaling is defective, we studied 3-mo-old female offspring of dams that were given EtOH during pregnancy compared with those from pair-fed and control dams. We performed an intraperitoneal pyruvate tolerance test, determined the phosphorylation status of hepatic phosphoinositide-dependent protein kinase-1 (PDK1), Akt, and PKCzeta before and after intravenous insulin bolus, and measured mRNA and in vivo acetylation of TRB3 (tribbles 3) and PTEN (phosphatase and tensin homolog deleted on chromosome ten) as well as the expression of the histone acetylase (HAT) PCAF (p300/CREB-binding protein-associated factor), histone deacetylase-1 (HDAC1), and HAT and HDAC activities. In EtOH compared with pair-fed and control offspring, basal and pyruvate-induced blood glucose was increased, insulin-induced PDK1, Akt, and PKCzeta phosphorylation was reduced, and expression of PTEN and TRB3 was increased while their acetylation status was decreased in association with increased HDAC and decreased HAT activities. Thus female adult rats prenatally exposed to EtOH have increased gluconeogenesis, reduced insulin signaling, and increased PTEN and TRB3 expression in the liver. In addition, PTEN and TRB3 are hypoacetylated, which can contribute to Akt-inhibiting activity. These results suggest that hepatic insulin resistance in rats prenatally exposed to EtOH is explained, at least in part, by increased PTEN and TRB3 activity due to both increased gene expression and reduced acetylation. PMID- 18385464 TI - Does ghrelin explain accelerated gastric emptying in the early stages of diabetes mellitus? AB - During the early stages of diabetes, gastric emptying is often accelerated, rather than delayed. The mechanism of accelerated gastric emptying in diabetes has not been fully studied. A recent study showed that plasma ghrelin levels were elevated in diabetes. As postprandial antropyloric coordination plays an important role in mediating solid gastric emptying, we hypothesize that the elevated plasma ghrelin levels increase postprandial antropyloric coordination to accelerate emptying in the early stages of diabetes. To test this hypothesis, rats were made diabetic by streptozotocin (STZ; 50 mg/kg) injection, and, 2 wk later, pre- and postprandial plasma ghrelin levels, antropyloric coordination, and solid gastric emptying were determined. In control rats, plasma ghrelin levels were immediately reduced after feeding. In contrast, plasma ghrelin levels remained within the fasted levels in STZ rats after feeding. In STZ rats, gastric emptying was significantly accelerated (77.4 +/- 3.2%, n = 6), compared with that of control rats (58.8 +/- 2.5%, n = 6, P < 0.05). Treatments with anti-ghrelin antibodies attenuated accelerated gastric emptying in STZ rats (50.1 +/- 3.5%, n = 6, P < 0.05), while having little effect in vehicle control rats. The incidence of postprandial antropyloric coordination was significantly increased in STZ rats, compared with that of control rats (P < 0.05). Treatments with anti-ghrelin antibodies suppressed this enhanced antropyloric coordination in STZ rats. Our study suggests that elevated endogenous ghrelin enhances antropyloric coordination, which accelerates gastric emptying in the early stages of diabetes. PMID- 18385465 TI - Exercise training normalizes enhanced glutamate-mediated sympathetic activation from the PVN in heart failure. AB - Exercise training (ExT) normalizes the increased sympathetic outflow in heart failure (HF), but the mechanisms are not known. We hypothesized that ExT would normalize the augmented glutamatergic mechanisms mediated by N-methyl-d-aspartic acid (NMDA) receptors within the paraventricular nucleus (PVN) that occur with HF. Four groups of rats were used: 1) sham-operated (Sham) sedentary (Sed), 2) Sham ExT, 3) HF Sed, and 4) HF ExT. HF was induced by left coronary artery ligation, and ExT consisted of 3 wk of treadmill running. In alpha-chloralose urethane-anesthetized rats, the increase in renal sympathetic nerve activity in response to the highest dose of NMDA (200 pmol) injected into the PVN in the HF Sed group was approximately twice that of the Sham Sed group. In the HF ExT group the response was not different from the Sham Sed and Sham ExT groups. Relative NMDA NR1 receptor subunit mRNA expression was 63% higher in the HF Sed group compared with the Sham Sed group but in the HF ExT group was not different from the Sham Sed and Sham ExT groups. NR1 receptor subunit protein expression was increased 87% in the HF Sed group compared with the Sham Sed group but in the HF ExT group was not significantly different from the Sham Sed and Sham ExT groups. Thus one mechanism by which ExT alleviates elevated sympathetic outflow in HF may be through normalization of glutamatergic mechanisms within the PVN. PMID- 18385466 TI - Blood flow responses in celiac and superior mesenteric arteries in the initial phase of digestion. AB - Blood flow (BF) responses in the celiac artery (CA) and superior mesenteric artery (SMA) during and immediately after a meal are poorly understood. We characterized postprandial BF responses in these arteries in the initial phase of digestion. After a baseline measurement in the overnight fasting state, healthy subjects ingested solid food (300 kcal) and water ad libitum within 5 min (4.6 +/ 0.2 min, means +/- SE), and then rested for 60 min in the postprandial state. Mean blood velocities (MBVs) in CA (n = 7) and SMA (n = 9) and mean arterial pressure (MAP) were measured throughout the procedure. The MAP was divided by the MBV to yield the resistance index (RI). The MBV in CA and SMA started increasing within a minute after beginning the meal. The MBV in CA rapidly reached its peak increase (60 +/- 8% change from baseline) at 5 +/- 1 min after the start of the meal, whereas the MBV in SMA gradually reached its peak increase (134 +/- 14%) at 41 +/- 4 min after the start of the meal, reflecting a decrease in the RI for both CA and SMA. These findings suggested an earlier increase in CA and SMA MBV, implying that the increase of BF in some parts of the small intestine precedes the arrival of chyme. PMID- 18385467 TI - Micturition-suppressing region in the periaqueductal gray of the mesencephalon of the cat. AB - The periaqueductal gray (PAG) of the mesencephalon has been implicated to be involved in the control of micturition. We investigated the micturition suppressing region in the PAG of the cat. Decerebrated 27 adult cats were used. A microelectrode was inserted stereotaxically into the PAG, and a region was searched where electrical stimulation suppressed isovolumetric bladder contractions. Simultaneous stimulation of the pontine micturition center (PMC) and micturition-suppressing region in the PAG was performed before and after an injection of bicuculline (GABA(A) blocker) into the PMC. The micturition suppressing region was found at the dorsolateral margin of the rostral PAG. Bladder contractions were not provoked by simultaneous stimulation of the PMC and micturition-suppressing region in the PAG. However, after bicuculline injection into the PMC, partial bladder contractions were provoked by simultaneous stimulation of the PMC and the micturition-suppressing region in the PAG. These results suggest that the dorsolateral margin of the rostral PAG includes the micturition-suppressing region that seems to have neural connections with the PMC. GABA is assumed to be one of the neurotransmitters that are involved in the PMC inhibition from the micturition-suppressing region in the PAG. PMID- 18385468 TI - Carrier-mediated urea transport across the mitochondrial membrane of an elasmobranch (Raja erinacea) and a teleost (Oncorhynchus mykiss) fish. AB - In osmoregulating teleost fish, urea is a minor nitrogen excretory product, whereas in osmoconforming marine elasmobranchs it serves as the major tissue organic solute and is retained at relatively high concentrations ( approximately 400 mmol/l). We tested the hypothesis that urea transport across liver mitochondria is carrier mediated in both teleost and elasmobranch fishes. Intact liver mitochondria in rainbow trout (Oncorhynchus mykiss) demonstrated two components of urea uptake, a linear component at high concentrations and a phloretin-sensitive saturable component [Michaelis constant (K(m)) = 0.58 mmol/l; maximal velocity (V(max)) = 0.12 mumol.h(-1).mg protein(-1)] at lower urea concentrations (<5 mmol/l). Similarly, analysis of urea uptake in mitochondria from the little skate (Raja erinacea) revealed a phloretin-sensitive saturable transport (K(m) = 0.34 mmol/l; V(max) = 0.054 mumol.h(-1).mg protein(-1)) at low urea concentrations (<5 mmol/l). Surprisingly, urea transport in skate, but not trout, was sensitive to a variety of classic ionophores and respiration inhibitors, suggesting cation sensitivity. Hence, urea transport was measured in the reverse direction using submitochondrial particles in skate. Transport kinetics, inhibitor response, and pH sensitivity were very similar in skate submitochondrial particle submitochondrial particles (K(m) = 0.65 mmol/l, V(max) = 0.058 mumol.h(-1).mg protein(-1)) relative to intact mitochondria. We conclude that urea influx and efflux in skate mitochondria is dependent, in part, on a bidirectional proton-sensitive mechanism similar to bacterial urea transporters and reminiscent of their ancestral origins. Rapid equilibration of urea across the mitochondrial membrane may be vital for cell osmoregulation (elasmobranch) or nitrogen waste excretion (teleost). PMID- 18385469 TI - Cardiovascular control during sleep: "Sleep-dependent changes in the coupling between heart period and blood pressure in human subjects," by Silvani et al. PMID- 18385470 TI - The high-fat-fed lean Zucker rat: a spontaneous isocaloric model of fat-induced insulin resistance associated with muscle GSK-3 overactivity. AB - High-fat feeding (HFF) is a well-accepted model for nutritionally-induced insulin resistance. The purpose of this investigation was to assess the metabolic responses of female lean Zucker rats provided regular chow (4% fat) or a high-fat chow (50% fat) for 15 wk. HFF rats spontaneously adjusted food intake so that daily caloric intake matched that of chow-fed (CF) controls. HFF animals consumed more (P < 0.05) calories from fat (31.9 +/- 1.2 vs. 2.4 +/- 0.2 kcal/day) and had significantly greater final body weights (280 +/- 10 vs. 250 +/- 5 g) and total visceral fat (24 +/- 3 vs. 10 +/- 1 g). Fasting plasma insulin was 2.3-fold elevated in HFF rats. Glucose tolerance (58%) and whole body insulin sensitivity (75%) were markedly impaired in HFF animals. In HFF plantaris muscle, in vivo insulin receptor beta-subunit (IR-beta) and insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation and phosphorylation of Akt Ser473 and glycogen synthase kinase-3beta (GSK-3beta) Ser9, relative to circulating insulin levels, were decreased by 40-59%. In vitro insulin-stimulated glucose transport in HFF soleus was decreased by 54%, as were IRS-1 tyrosine phosphorylation (26%) and phosphorylation of Akt Ser473 (38%) and GSK-3beta Ser9 (25%), the latter indicative of GSK-3 overactivity. GSK-3 inhibition in HFF soleus using CT98014 increased insulin-stimulated glucose transport (28%), IRS-1 tyrosine phosphorylation (28%) and phosphorylation of Akt Ser473 (38%) and GSK-3beta Ser9 (48%). In summary, the female lean Zucker rat fed a high-fat diet represents an isocaloric model of nutritionally-induced insulin resistance associated with moderate visceral fat gain, hyperinsulinemia, and impairments of skeletal muscle insulin-signaling functionality, including GSK-3beta overactivity. PMID- 18385471 TI - Relationship between low magnesium status and TRPM6 expression in the kidney and large intestine. AB - The body maintains Mg(2+) homeostasis by renal and intestinal (re)absorption. However, the molecular mechanisms that mediate transepithelial Mg(2+) transport are largely unknown. Transient receptor potential melastatin 6 (TRPM6) was recently identified and shown to function in active epithelial Mg(2+) transport in intestine and kidney. To define the relationship between Mg(2+) status and TRPM6 expression, we used two models of hypomagnesemia: 1) C57BL/6J mice fed a mildly or severely Mg(2+)-deficient diet, and 2) mice selected for either low (MgL) or high (MgH) erythrocyte and plasma Mg(2+) status. In addition, the mice were subjected to a severely Mg(2+)-deficient diet. Our results show that C57BL/6J mice fed a severely Mg(2+)-deficient diet developed hypomagnesemia and hypomagnesuria and showed increased TRPM6 expression in kidney and intestine. When fed a Mg(2+)-adequate diet, MgL mice presented hypomagnesemia and hypermagnesuria, and lower kidney and intestinal TRPM6 expression, compared with MgH mice. A severely Mg(2+)-deficient diet led to hypomagnesemia and hypomagnesuria in both strains. Furthermore, this diet induced kidney TRPM6 expression in MgL mice, but not in MgH mice. In conclusion, as shown in C57BL/6J mice, dietary Mg(2+)-restriction results in increased Mg(2+) (re)absorption, which is correlated with increased TRPM6 expression. In MgL and MgH mice, the inherited Mg(2+) status is linked to different TRPM6 expression. The MgL and MgH mice respond differently to a low-Mg(2+) diet with regard to TRPM6 expression in the kidney, consistent with genetic factors contributing to the regulation of cellular Mg(2+) levels. Further studies of these mice strains could improve our understanding of the genetics of Mg(2+) homeostasis. PMID- 18385472 TI - Tachykinin neurokinin 3 receptor signaling in cholecystokinin-elicited release of oxytocin and vasopressin. AB - Neurokinin 3 receptor (NK3R) signaling has an integral role in the stimulated oxytocin (OT) and vasopressin (VP) release in response to hyperosmolarity and hypotension. Peripheral injections of cholecystokinin (CCK) receptor agonists for the CCK-A (sulfated CCK-8) and CCK-B (nonsulfated CCK-8) receptors elicit an OT release in rat. It is unknown whether NK3R contributes to this endocrine response. Freely behaving male rats were administered an intraventricular pretreatment of 250 or 500 pmol of SB-222200, a specific NK3R antagonist, or 0.15 M NaCl before an intraperitoneal or intravenous injection of CCK-8 (nonsulfated or sulfated) or 0.15 M NaCl. Blood samples were taken before intraventricular treatment and 15 min after intraperitoneal or intravenous injection, and plasma samples were assayed for OT and VP concentration. Intraperitoneal injection of both nonsulfated and sulfated CCK-8 significantly increased plasma OT levels and had no effect on plasma VP levels. Intravenous injection of sulfated CCK-8 stimulated an increase in plasma OT levels and did not alter plasma VP levels. However, intravenous injection of nonsulfated CCK-8 stimulated a significant increase in plasma levels of both OT and VP. No other studies have demonstrated CCK-8-stimulated release of VP in rat. NK3R antagonist did not alter baseline levels of either hormone. However, pretreatment of NK3R antagonist significantly blocked the CCK-stimulated release of OT in all CCK treatment groups and blocked VP release in response to intravenous injection of nonsulfated CCK-8. Therefore, central NK3R signaling is required for OT and VP release in response to CCK administration. PMID- 18385473 TI - The neural circuit for tone-specific plasticity in the auditory system elicited by conditioning. PMID- 18385475 TI - Contextual memory deficits observed in mice overexpressing small conductance Ca2+ activated K+ type 2 (KCa2.2, SK2) channels are caused by an encoding deficit. AB - Hippocampal-dependent synaptic plasticity and memory are modulated by apamin sensitive small conductance Ca2+-activated K+ (SK) channels. Transgenic mice overexpressing SK2 channels (SK2+/T mice) exhibit marked deficits in hippocampal memory and synaptic plasticity, as previously reported. Here, we examined whether SK2 overexpression affects the encoding or retention of contextual memory. Compared with wild-type littermates, SK2+/T mice exhibited significantly less context-dependent freezing 10 min and 24 h after conditioning. Interestingly, this contextual memory impairment was eliminated if SK2+/T mice were permitted longer pre-exposure to the conditioning chamber. These data support converging evidence that SK2 channels restrict the encoding of hippocampal memory. PMID- 18385476 TI - Retrograde amnesia for visual memories after hippocampal damage in rats. AB - It is generally believed that the hippocampus is not required for simple discrimination learning. However, a small number of studies have shown that hippocampus damage impairs retention of a previously learned visual discrimination task. We propose that, although simple discrimination learning may proceed in the absence of the hippocampus, it plays an important role in this type of learning when it is intact. In order to test the role of the hippocampus in simple discrimination learning, we performed a series of experiments utilizing a two-choice picture discrimination task. Our experiments confirm that rats readily learn simple two-choice picture discriminations after hippocampus damage. However, if such discriminations are first learned while the hippocampus is intact, subsequent hippocampus damage causes severe retrograde amnesia for the discriminations. Furthermore, retrograde amnesia for simple picture discriminations was equally severe when the interval between training and damage was 1 d or 60 d; remote picture memories are not spared. Similarly, the rule or schema underlying a recently or remotely acquired picture discrimination learning set was lost after hippocampus damage. The severity of retrograde amnesia for simple picture discriminations is negatively correlated with the volume of spared hippocampus tissue. Thus, the hippocampus plays an essential role in long-term memories supporting simple picture discriminations. PMID- 18385477 TI - Sustained increase in hippocampal sharp-wave ripple activity during slow-wave sleep after learning. AB - High-frequency oscillations, known as sharp-wave/ripple (SPW-R) complexes occurring in hippocampus during slow-wave sleep (SWS), have been proposed to promote synaptic plasticity necessary for memory consolidation. We recorded sleep for 3 h after rats were trained on an odor-reward association task. Learning resulted in an increased number SPW-Rs during the first hour of post-learning SWS. The magnitude of ripple events and their duration were also elevated for up to 2 h after the newly formed memory. Rats that did not learn the discrimination during the training session did not show any change in SPW-Rs. Successful retrieval from remote memory was likewise accompanied by an increase in SPW-R density and magnitude, relative to the previously recorded baseline, but the effects were much shorter lasting and did not include increases in ripple duration and amplitude. A short-lasting increase of ripple activity was also observed when rats were rewarded for performing a motor component of the task only. There were no increases in ripple activity after habituation to the experimental environment. These experiments show that the characteristics of hippocampal high-frequency oscillations during SWS are affected by prior behavioral experience. Associative learning induces robust and sustained (up to 2 h) changes in several SPW-R characteristics, while after retrieval from remote memory or performance of a well-trained procedural aspect of the task, only transient changes in ripple density were induced. PMID- 18385478 TI - Nitric oxide is involved in appetitive but not aversive olfactory learning in the land mollusk Limax valentianus. AB - The land slug Limax performs both aversive and appetitive olfactory learning, and we investigated neurotransmitters involved in each type of learning. Slugs were conditioned by presenting a vegetable juice (appetitive conditioning) or a mixture of vegetable juice and quinidine (aversive conditioning), and the latency to reach the juice became shorter (appetitive conditioning) or longer (aversive conditioning) after conditioning. L-NAME injected either before conditioning or testing blocked the reduction in latency in appetitive conditioning but had no significant effects in aversive conditioning. 5,7-dihydroxytryptamine had no significant effects in appetitive conditioning. These results suggest different mechanisms for appetitive and aversive learning. PMID- 18385479 TI - Drosophila ortholog of succinyl-CoA synthetase {beta} subunit: a novel modulator of Drosophila KCNQ channels. AB - Voltage-gated KCNQ potassium channels are responsible for slowly activating potassium currents in heart, brain, and other tissues. Functional defects of KCNQ channels are linked with many diseases, including epilepsy and cardiac arrhythmias. Therefore KCNQ potassium channels have been widely studied, especially in the CNS. We have identified Drosophila CG11963, which encodes a protein orthologous to the beta subunit of mammalian succinyl-CoA synthetase (SCS, also known as succinate thiokinase), as a novel modulator of Drosophila KCNQ channels. Direct interaction of CG11963 and dKCNQ was demonstrated by yeast two-hybrid screen and coimmunoprecipitation. Cell surface biotinylation experiments further confirmed that CG11963 resides on the plasma membrane of tsA 201 cells. Coexpression of CG11963 with dKCNQ shifts the conductance-voltage (G V) relationship of dKCNQ channels to more positive membrane potentials in Chinese hamster ovary (CHO) cells. Moreover, directly dialyzing glutathione S-transferase fusion CG11963 protein into CHO cells also shifts the dKCNQ G-V curve rightward. The effect of CG11963 persists in the presence of 1 mM adenosine triphosphate (ATP), a substrate of SCS. Taken together, our data define CG11963 as a new dKCNQ binding protein capable of modulating the properties of the channel. Our evidence suggests that this modulation is mediated by direct interaction of CG11963 with the channel and is not dependent on ATP. PMID- 18385480 TI - Spike-firing resonance in hypoglossal motoneurons. AB - During an inspiration the output of hypoglossal (XII) motoneurons (HMs) in vitro is characterized by synchronous oscillatory firing in the 20- to 40-Hz range. To maintain synchronicity it is important that the cells fire with high reliability and precision. It is not known whether the intrinsic properties of HMs are tuned to maintain synchronicity when stimulated with time-varying inputs. We intracellularly recorded from HMs in an in vitro brain stem slice preparation from juvenile mice. Cells were held at or near spike threshold and were stimulated with steady or swept sine-wave current functions (10-s duration; 0- to 40-Hz range). Peristimulus time histograms were constructed from spike times based on threshold crossings. Synaptic transmission was suppressed by including blockers of GABAergic, glycinergic, and glutamatergic neurotransmission in the bath solution. Cells responded to sine-wave stimulation with bursts of action potentials at low (<3- to 5-Hz) sine-wave frequency, whereas they phase-locked 1:1 to the stimulus at intermediate frequencies (3-25 Hz). Beyond the 1:1 frequency range cells were able to phase-lock to subharmonics (1:2, 1:3, or 1:4) of the input frequency. The 1:1 phase-locking range increased with increasing stimulus amplitude and membrane depolarization. Reliability and spike-timing precision were highest when the cells phase-locked 1:1 to the stimulus. Our findings suggest that the coding of time-varying inspiratory synaptic inputs by individual HMs is most reliable and precise at frequencies that are generally lower than the frequency of the synchronous inspiratory oscillatory activity recorded from the XII nerve. PMID- 18385481 TI - Na+,K+,2Cl- cotransport and intracellular chloride regulation in rat primary sensory neurons: thermodynamic and kinetic aspects. AB - Adult primary afferent neurons are depolarized by GABA throughout their entire surface, including their somata located in dorsal root ganglia (DRG). Primary afferent depolarization (PAD) mediated by GABA released from spinal interneurons determines presynaptic inhibition, a key mechanism in somatosensory processing. The depolarization is due to Cl(-) efflux through GABA(A) channels; the outward Cl(-) gradient is generated by a Na+,K+,2Cl(-) cotransporter (NKCC) as first established in amphibians. Using fluorescence imaging microscopy we measured [Cl( )]i and cell water volume (CWV) in dissociated rat DRG cells (P0-P21) loaded with N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide and calcein, respectively. Basal [Cl(-)]i was 44.2 +/- 1.2 mM (mean +/- SE), Cl(-) equilibrium potential (E Cl) was -27.0 +/- 0.7 mV (n = 75). This [Cl(-)]i is about four times higher than electrochemical equilibrium. On isosmotic removal of external Cl(-), cells lost Cl(-) and shrank. On returning to control solution, cells reaccumulated Cl(-) and recovered CWV. Cl(-) reaccumulation had Na+-dependent (SDC) and Na+-independent (SIC) components. The SIC stabilized at [Cl(-)]i = 13.2 +/- 1.2 mM, suggesting that it was passive (E(Cl) = -60.5 +/- 3 mV). Bumetanide blocked CWV recovery and most (65%) of the SDC (IC50 = 5.7 microM), indicating that both were mediated by NKCC. Active Cl(-) uptake fell with increasing [Cl(-)]i and became negligible when [Cl(-)]i reached basal levels. The kinetics of active Cl(-) uptake suggests a negative feedback system in which intracellular Cl(-)regulates its own influx thereby keeping [Cl(-)]i constant, above electrochemical equilibrium but below the value that would attain if NKCC reached thermodynamic equilibrium. PMID- 18385482 TI - Increasing extracellular potassium results in subthalamic neuron activity resembling that seen in a 6-hydroxydopamine lesion. AB - Abnormal neuronal activity in the subthalamic nucleus (STN) plays a crucial role in the pathophysiology of Parkinson's disease (PD). Although altered extracellular potassium concentration ([K+]o) and sensitivity to [K+]o modulates neuronal activity, little is known about the potassium balance in the healthy and diseased STN. In vivo measurements of [K+]o using ion-selective electrodes demonstrated a twofold increase in the decay time constant of lesion-induced [K+]o transients in the STN of adult Wistar rats with a unilateral 6 hydroxydopamine (6-OHDA) median forebrain bundle lesion, employed as a model of PD, compared with nonlesioned rats. Various [K+]o concentrations (1.5-12.5 mM) were applied to in vitro slice preparations of three experimental groups of STN slices from nonlesioned control rats, ipsilateral hemispheres, and contralateral hemispheres of lesioned rats. The majority of STN neurons of nonlesioned rats and in slices contralateral to the lesion fired spontaneously, predominantly in a regular pattern, whereas those in slices ipsilateral to the lesion fired more irregularly or even in bursts. Experimentally increased [K+]o led to an increase in the number of spontaneously firing neurons and action potential firing rates in all groups. This was accompanied by a decrease in the amplitude of post spike afterhyperpolarization (AHP) and the amplitude and duration of the posttrain AHP. Lesion effects in ipsilateral neurons at physiological [K+]o resembled the effects of elevated [K+]o in nonlesioned rats. Our data suggest that changed potassium sensitivity due to conductivity alterations and delayed clearance may be critical for shaping STN activity in parkinsonian states. PMID- 18385483 TI - Distinct cortical anatomy linked to subregions of the medial temporal lobe revealed by intrinsic functional connectivity. AB - The hippocampus and adjacent cortical structures in the medial temporal lobe (MTL) contribute to memory through interactions with distributed brain areas. Studies of monkey and rodent anatomy suggest that parallel pathways converge on distinct subregions of the MTL. To explore the cortical areas linked to subregions of the MTL in humans, we examined cortico-cortical and hippocampal cortical correlations using high-resolution, functional connectivity analysis in 100 individuals. MTL seed regions extended along the anterior to posterior axis and included hippocampus and adjacent structures. Results revealed two separate brain pathways that correlated with distinct subregions within the MTL. The body of the hippocampus and posterior parahippocampal cortex correlated with lateral parietal cortex, regions along the posterior midline including posterior cingulate and retrosplenial cortex, and ventral medial prefrontal cortex. By contrast, anterior hippocampus and the perirhinal/entorhinal cortices correlated with distinct regions in the lateral temporal cortex extending into the temporal pole. The present results are largely consistent with known connectivity in the monkey and provide a novel task-independent dissociation of the parallel pathways supporting the MTL memory system in humans. The cortical pathways include regions that have undergone considerable areal expansion in humans, providing insight into how the MTL memory system has evolved to support a diverse array of cognitive domains. PMID- 18385484 TI - Mechanisms of neuromodulation as dissected using Sr2+ at motor nerve endings. AB - The use of binomial analysis as a tool for determining the sites of action of neuromodulators may be complicated by the nonuniformity of release probability. One of the potential sources for nonuniformity of release probability is the presence of multiple forms of synaptotagmins, the Ca2+ sensors responsible for triggering vesicular exocytosis. In this study we have used Sr2+, an ion whose actions may be restricted to a subpopulation of synaptotagmins, in an attempt to obtain meaningful estimates of the binomial parameters p (the probability of evoked acetylcholine [Ach] release) and n (the immediate available store of ACh quanta, whereby m = np). In contrast to results in Ca2+ solutions, binomial analysis of Sr2+-dependent release reveals a dramatically reduced dependence of n on extracellular Sr2+ concentrations. In Sr2+ solutions, blockade of potassium channels with 3,4-diaminopyridine increased m by an exclusive increase in p, whereas treatment with phorbol ester increased m solely by effects on n. The cyclic adenosine monophosphate (cAMP) analogue CPT-cAMP increased m by increasing both n and p. The effect of CPT-cAMP on p but not on n was blocked by protein kinase A (PKA) inhibitors, whereas the effect on n was mimicked by 8-CPT-2'-O-Me cAMP, a selective agonist for exchange protein directly activated by cAMP, otherwise known as the cAMP-sensitive guanine nucleotide-exchange protein. The results demonstrate both the utility of the binomial distribution in Sr2+ solutions and the dual effects of cyclic AMP on both PKA-dependent and PKA independent processes at the amphibian neuromuscular junction. PMID- 18385486 TI - Physiology and morphology of shared and specialized spinal interneurons for locomotion and scratching. AB - Distinct types of rhythmic movements that use the same muscles are typically generated largely by shared multifunctional neurons in invertebrates, but less is known for vertebrates. Evidence suggests that locomotion and scratching are produced partly by shared spinal cord interneuronal circuity, although direct evidence with intracellular recording has been lacking. Here, spinal interneurons were recorded intracellularly during fictive swimming and fictive scratching in vivo and filled with Neurobiotin. Some interneurons that were rhythmically activated during both swimming and scratching had axon terminal arborizations in the ventral horn of the hindlimb enlargement, indicating their likely contribution to hindlimb motor outputs during both behaviors. We previously described a morphological group of spinal interneurons ("transverse interneurons" or T neurons) that were rhythmically activated during all forms of fictive scratching at higher peak firing rates and with larger membrane potential oscillations than scratch-activated spinal interneurons with different dendritic orientations. The current study demonstrates that T neurons are activated during both swimming and scratching and thus are components of the shared circuitry. Many spinal interneurons activated during fictive scratching are also activated during fictive swimming (scratch/swim neurons), but others are suppressed during swimming (scratch-specialized neurons). The current study demonstrates that some scratch-specialized neurons receive strong and long-lasting hyperpolarizing inhibition during fictive swimming and are also morphologically distinct from T neurons. Thus this study indicates that locomotion and scratching are produced by a combination of shared and dedicated interneurons whose physiological and morphological properties are beginning to be revealed. PMID- 18385485 TI - A theory of the dual pathways for smooth pursuit based on dynamic gain control. AB - The smooth pursuit eye movement (SPEM) system is much more sensitive to target motion perturbations during pursuit than during fixation. This sensitivity is commonly attributed to a dynamic gain control mechanism. Neither the neural substrate nor the functional architecture for this gain control has been fully revealed. There are at least two cortical areas that crucially contribute to smooth pursuit and are therefore eligible sites for dynamic gain control: the medial superior temporal area (MST) and the pursuit area of the frontal eye fields (FEFs), which both project to brain stem premotor structures via parallel pathways. The aim of this study was to develop a model of smooth pursuit based on behavioral, anatomical, and neurophysiological results to account for nonlinear dynamic gain control. Using a behavioral paradigm in humans consisting of a sinusoidal oscillation (4 Hz, +/-8 degrees/s) superimposed on a constant velocity target motion (0-24 degrees/s), we were able to identify relevant gain control parameters in the model. A salient feature of our model is the emergence of two parallel pathways from higher visual cortical to lower motor areas in the brain stem that correspond to the MST and FEF pathways. Detailed analysis of the model revealed that one pathway mainly carries eye velocity related signals, whereas the other is associated mostly with eye acceleration. From comparison with known neurophysiological results we conclude that the dynamic gain control can be attributed to the FEF pathway, whereas the MST pathway serves as the basic circuit for maintaining an ongoing SPEM. PMID- 18385487 TI - Descending projections from auditory cortex modulate sensitivity in the midbrain to cues for spatial position. AB - The function of the profuse descending innervation from the auditory cortex is largely unknown; however, recent studies have demonstrated that focal stimulation of auditory cortex effects frequency tuning curves, duration tuning, and other auditory parameters in the inferior colliculus. Here we demonstrate that, in an anesthetized guinea pig, nonfocal deactivation of the auditory cortex alters the sensitivity of populations of neurons in the inferior colliculus (IC) to one of the major cues for the localization of sound in space, interaural level differences (ILDs). Primary and secondary auditory cortical areas were inactivated by cooling. The ILD functions of 46% of IC cells changed when the cortex was inactivated. In extreme cases, the ILD functions changed from monotonic to nonmonotonic during cooling and vice versa. Eight percent of the cells became unresponsive after deactivation of the auditory cortex. Deactivation of the cortex has previously been shown to alter the maximum spike count of cells in the IC; the change in normalized ILD functions is shown to be separate from this effect. In some cases, the ILD function changed shape when there was no change in the maximum spike count and in other cases there was no change in the shape of the ILD function even though there was a large change in the maximum spike count. Overall, the sensitivity of the IC neural population to ILD is radically altered by the corticofugal pathway. PMID- 18385488 TI - Integrating adult learning principles into training for public health practice. AB - Providing training and planned learning experiences to practitioners and community partners is fundamental to effective public health. The extent to which principles of adult learning currently guide such training is unknown and likely varies widely. The purpose of this article is to introduce five principles of adult learning and discuss how each can be applied in assessing trainee needs, planning and delivering training, and evaluating training processes and outcomes. Training guided by these principles should facilitate adult learning, collaborative efforts, and mutual respect between agencies, practitioners, and community partners. PMID- 18385489 TI - Designing a cardiovascular disease prevention web site for Latinos: qualitative community feedback. AB - Latinos are the largest minority group in the United States, yet there is currently a lack of Web sites that focus on Latino health. This article shares qualitative research results obtained from Latinos, with a focus on creating a culturally sensitive, bilingual, interactive, computer-based cardiovascular disease risk assessment and behavioral goal-setting program. This project is a substudy of the Latinos Using Cardio Health Actions to Reduce Risk (LUCHAR) grant, a 5-year trial, funded by the National Heart, Lung, and Blood Institute, whose primary aim is the primary prevention of cardiovascular disease in Latinos. Ten focus groups, five in English and five in Spanish, were conducted at six community sites with trained, bilingual facilitators in Denver, Colorado. Results from the focus groups are used to make recommendations on how to develop a culturally sensitive cardiovascular health care Web site for Latinos. PMID- 18385490 TI - An inland and a coastal population of the Mediterranean xero-halophyte species Atriplex halimus L. differ in their ability to accumulate proline and glycinebetaine in response to salinity and water stress. AB - Soil salinity and drought compromise water uptake and lead to osmotic adjustment in xero-halophyte plant species. These important environmental constraints may also have specific effects on plant physiology. Stress-induced accumulation of osmocompatible solutes was analysed in two Tunisian populations of the Mediteranean shrub Atriplex halimus L.-plants originating from a salt-affected coastal site (Monastir) or from a non-saline semi-arid area (Sbikha)-were exposed to nutrient solution containing either low (40 mM) or high (160 mM) doses of NaCl or 15% polyethylene glycol. The low NaCl dose stimulated plant growth in both populations. Plants from Monastir were more resistant to high salinity and exhibited a greater ability to produce glycinebetaine in response to salt stress. Conversely, plants from Sbikha were more resistant to water stress and displayed a higher rate of proline accumulation. Proline accumulated as early as 24 h after stress imposition and such accumulation was reversible. By contrast, glycinebetaine concentration culminated after 10 d of stress and did not decrease after the stress relief. The highest salt resistance of Monastir plants was not due to a lower rate of Na(+) absorption; plants from this population exhibited a higher stomatal conductance and a prodigal water-use strategy leading to lower water-use efficiency than plants from Sbikha. Exogenous application of proline (1 mM) improved the level of drought resistance in Monastir plants through a decrease in oxidative stress quantified by the malondialdehyde concentration, while the exogenous application of glycinebetaine improved the salinity resistance of Sbikha plants through a positive effect on photosystem II efficiency. PMID- 18385492 TI - Clarification of funding of early lung cancer study. PMID- 18385491 TI - Full disclosure and the funding of biomedical research. PMID- 18385493 TI - Videos in clinical medicine. Female urethral catheterization. PMID- 18385494 TI - Images in clinical medicine. Ascariasis. PMID- 18385495 TI - Regulating off-label drug use--rethinking the role of the FDA. PMID- 18385496 TI - Association between obesity during pregnancy and increased use of health care. AB - BACKGROUND: In the United States, obesity during pregnancy is common and increases obstetrical risks. An estimate of the increase in use of health care services associated with obesity during pregnancy is needed. METHODS: We used electronic data systems of a large U.S. group-practice health maintenance organization to identify 13,442 pregnancies among women 18 years of age or older at the time of conception that resulted in live births or stillbirths. The study period was between January 1, 2000, and December 31, 2004. We assessed associations between measures of use of health care services and body-mass index (BMI, defined as the weight in kilograms divided by the square of the height in meters) before pregnancy or in early pregnancy. The women were categorized as underweight (BMI <18.5), normal (BMI 18.5 to 24.9), overweight (BMI 25.0 to 29.9), obese (BMI 30.0 to 34.9), very obese (BMI 35.0 to 39.9), or extremely obese (BMI > or =40.0). The primary outcome was the mean length of hospital stay for delivery. RESULTS: After adjustment for age, race or ethnic group, level of education, and parity, the mean (+/-SE) length of hospital stay for delivery was significantly (P<0.05) greater among women who were overweight (3.7+/-0.1 days), obese (4.0+/-0.1 days), very obese (4.1+/-0.1 days), and extremely obese (4.4+/ 0.1 days) than among women with normal BMI (3.6+/-0.1 days). A higher-than-normal BMI was associated with significantly more prenatal fetal tests, obstetrical ultrasonographic examinations, medications dispensed from the outpatient pharmacy, telephone calls to the department of obstetrics and gynecology, and prenatal visits with physicians. A higher-than-normal BMI was also associated with significantly fewer prenatal visits with nurse practitioners and physician assistants. Most of the increase in length of stay associated with higher BMI was related to increased rates of cesarean delivery and obesity-related high-risk conditions. CONCLUSIONS: Obesity during pregnancy is associated with increased use of health care services. PMID- 18385497 TI - Gene identification for the cblD defect of vitamin B12 metabolism. AB - BACKGROUND: Vitamin B12 (cobalamin) is an essential cofactor in several metabolic pathways. Intracellular conversion of cobalamin to its two coenzymes, adenosylcobalamin in mitochondria and methylcobalamin in the cytoplasm, is necessary for the homeostasis of methylmalonic acid and homocysteine. Nine defects of intracellular cobalamin metabolism have been defined by means of somatic complementation analysis. One of these defects, the cblD defect, can cause isolated methylmalonic aciduria, isolated homocystinuria, or both. Affected persons present with multisystem clinical abnormalities, including developmental, hematologic, neurologic, and metabolic findings. The gene responsible for the cblD defect has not been identified. METHODS: We studied seven patients with the cblD defect, and skin fibroblasts from each were investigated in cell culture. Microcell-mediated chromosome transfer and refined genetic mapping were used to localize the responsible gene. This gene was transfected into cblD fibroblasts to test for the rescue of adenosylcobalamin and methylcobalamin synthesis. RESULTS: The cblD gene was localized to human chromosome 2q23.2, and a candidate gene, designated MMADHC (methylmalonic aciduria, cblD type, and homocystinuria), was identified in this region. Transfection of wild-type MMADHC rescued the cellular phenotype, and the functional importance of mutant alleles was shown by means of transfection with mutant constructs. The predicted MMADHC protein has sequence homology with a bacterial ATP-binding cassette transporter and contains a putative cobalamin binding motif and a putative mitochondrial targeting sequence. CONCLUSIONS: Mutations in a gene we designated MMADHC are responsible for the cblD defect in vitamin B12 metabolism. Various mutations are associated with each of the three biochemical phenotypes of the disorder. PMID- 18385498 TI - Treatment of vulvar intraepithelial neoplasia with topical imiquimod. AB - BACKGROUND: Alternatives to surgery are needed for the treatment of vulvar intraepithelial neoplasia. We investigated the effectiveness of imiquimod 5% cream, a topical immune-response modulator, for the treatment of this condition. METHODS: Fifty-two patients with grade 2 or 3 vulvar intraepithelial neoplasia were randomly assigned to receive either imiquimod or placebo, applied twice weekly for 16 weeks. The primary outcome was a reduction of more than 25% in lesion size at 20 weeks. Secondary outcomes were histologic regression, clearance of human papillomavirus (HPV) from the lesion, changes in immune cells in the epidermis and dermis of the vulva, relief of symptoms, improvement of quality of life, and durability of response. Reduction in lesion size was classified as complete response (elimination), strong partial response (76 to 99% reduction), weak partial response (26 to 75% reduction), or no response (< or =25% reduction). The follow-up period was 12 months. RESULTS: Lesion size was reduced by more than 25% at 20 weeks in 21 of the 26 patients (81%) treated with imiquimod and in none of those treated with placebo (P<0.001). Histologic regression was significantly greater in the imiquimod group than in the placebo group (P<0.001). At baseline, 50 patients (96%) tested positive for HPV DNA. HPV cleared from the lesion in 15 patients in the imiquimod group (58%), as compared with 2 in the placebo group (8%) (P<0.001). The number of immune epidermal cells increased significantly and the number of immune dermal cells decreased significantly with imiquimod as compared with placebo. Imiquimod reduced pruritus and pain at 20 weeks (P=0.008 and P=0.004, respectively) and at 12 months (P=0.04 and P=0.02, respectively). The lesion progressed to invasion (to a depth of <1 mm) in 3 of 49 patients (6%) followed for 12 months (2 in the placebo group and 1 in the imiquimod group). Nine patients, all treated with imiquimod, had a complete response at 20 weeks and remained free from disease at 12 months. CONCLUSIONS: Imiquimod is effective in the treatment of vulvar intraepithelial neoplasia. (Current Controlled Trials number, ISRCTN11290871 [controlled trials.com].). PMID- 18385499 TI - Clinical practice. Osteoporosis in men. PMID- 18385500 TI - Atopic dermatitis. PMID- 18385501 TI - Images in clinical medicine. Vestibular papillomatosis. PMID- 18385502 TI - Clinical problem-solving. An unintended consequence--a 79-year-old man with a 5 month history of fatigue and 20-lb (9-kg) weight loss presented to his local physician. PMID- 18385503 TI - Options for slowing the growth of health care costs. PMID- 18385504 TI - Platelet-activating factor, PAF acetylhydrolase, and anaphylaxis. PMID- 18385505 TI - Telbivudine versus lamivudine in patients with chronic hepatitis B. PMID- 18385506 TI - Patent foramen ovale and cryptogenic stroke. PMID- 18385507 TI - Adolescent overweight and coronary heart disease. PMID- 18385508 TI - Cochlear implants. PMID- 18385509 TI - Hurricane Katrina and disaster preparedness. PMID- 18385510 TI - Microembolism during foam sclerotherapy of varicose veins. PMID- 18385511 TI - Statistical issues in the use of the comet assay. AB - The comet or single-cell gel electrophoresis assay is now widely used in regulatory, mechanistic and biomonitoring studies using a range of in vitro and in vivo systems. Each of these has issues associated with the experimental design which determine to a large extent the statistical analyses than can be used. A key concept is that the experimental unit is the smallest 'amount' of experimental material that can be randomly assigned to a treatment: the animal for in vivo studies and the culture for in vitro studies. Biomonitoring studies, being observational rather than experimental, are vulnerable to confounding and biases. Critical factors in any statistical analysis include the identification of suitable end points, the choice of measure to represent the distribution of the comet end point in a sample of cells, estimates of variability between experimental units and the identification of the size of effects that could be considered biologically important. Power and sample size calculations can be used in conjunction with this information to identify optimum experimental sizes and provide help in combining the results of statistical analyses with other information to aid interpretation. Interpretation based upon the size of effects and their confidence intervals is preferred to that based solely upon statistical significance tests. Statistical issues associated with the design and subsequent analyses of current validation studies for the comet assay include the identification of acceptable levels of intra- and inter-laboratory repeatability and reproducibility and criteria for dichotomizing results into positive or negative. PMID- 18385512 TI - HOPS proofreads the trans-SNARE complex for yeast vacuole fusion. AB - The fusion of yeast vacuoles, like other organelles, requires a Rab-family guanosine triphosphatase (Ypt7p), a Rab effector and Sec1/Munc18 (SM) complex termed HOPS (homotypic fusion and vacuole protein sorting), and soluble N ethylmaleimide-sensitive factor attachment protein receptors (SNAREs). The central 0-layer of the four bundled vacuolar SNAREs requires the wild-type three glutaminyl (Q) and one arginyl (R) residues for optimal fusion. Alterations of this layer dramatically increase the K(m) value for SNAREs to assemble trans SNARE complexes and to fuse. We now find that added purified HOPS complex strongly suppresses the fusion of vacuoles bearing 0-layer alterations, but it has little effect on the fusion of vacuoles with wild-type SNAREs. HOPS proofreads at two levels, inhibiting the formation of trans-SNARE complexes with altered 0-layers and suppressing the ability of these mismatched 0-layer trans SNARE complexes to support membrane fusion. HOPS proofreading also extends to other parts of the SNARE complex, because it suppresses the fusion of trans-SNARE complexes formed without the N-terminal Phox homology domain of Vam7p (Q(c)). Unlike some other SM proteins, HOPS proofreading does not require the Vam3p (Q(a)) N-terminal domain. HOPS thus proofreads SNARE domain and N-terminal domain structures and regulates the fusion capacity of trans-SNARE complexes, only allowing full function for wild-type SNARE configurations. This is the most direct evidence to date that HOPS is directly involved in the fusion event. PMID- 18385513 TI - A supraphysiological nuclear export signal is required for parvovirus nuclear export. AB - CRM1 exports proteins that carry a short leucine-rich peptide signal, the nuclear export signal (NES), from the nucleus. Regular NESs must have low affinity for CRM1 to function optimally. We previously generated artificial NESs with higher affinities for CRM1, termed supraphysiological NESs. Here we identify a supraphysiological NES in an endogenous protein, the NS2 protein of parvovirus Minute Virus of Mice (MVM). NS2 interacts with CRM1 without the requirement of RanGTP, whereas addition of RanGTP renders the complex highly stable. Mutation of a single hydrophobic residue that inactivates regular NESs lowers the affinity of the NS2 NES for CRM1 from supraphysiological to regular. Mutant MVM harboring this regular NES is compromised in viral nuclear export and productivity. In virus-infected mouse fibroblasts we observe colocalization of NS2, CRM1 and mature virions, which is dependent on the supraphysiological NS2 NES. We conclude that supraphysiological NESs exist in nature and that the supraphysiological NS2 NES has a critical role in active nuclear export of mature MVM particles before cell lysis. PMID- 18385514 TI - RAB-11 permissively regulates spindle alignment by modulating metaphase microtubule dynamics in Caenorhabditis elegans early embryos. AB - Alignment of the mitotic spindle along a preformed axis of polarity is crucial for generating cell diversity in many organisms, yet little is known about the role of the endomembrane system in this process. RAB-11 is a small GTPase enriched in recycling endosomes. When we depleted RAB-11 by RNAi in Caenorhabditis elegans, the spindle of the one-cell embryo failed to align along the axis of polarity in metaphase and underwent violent movements in anaphase. The distance between astral microtubules ends and the anterior cortex was significantly increased in rab-11(RNAi) embryos specifically during metaphase, possibly accounting for the observed spindle alignment defects. Additionally, we found that normal ER morphology requires functional RAB-11, particularly during metaphase. We hypothesize that RAB-11, in conjunction with the ER, acts to regulate cell cycle-specific changes in astral microtubule length to ensure proper spindle alignment in Caenorhabditis elegans early embryos. PMID- 18385515 TI - Novel interactions of ESCRT-III with LIP5 and VPS4 and their implications for ESCRT-III disassembly. AB - The AAA+ ATPase VPS4 plays an essential role in multivesicular body biogenesis and is thought to act by disassembling ESCRT-III complexes. VPS4 oligomerization and ATPase activity are promoted by binding to LIP5. LIP5 also binds to the ESCRT III like protein CHMP5/hVps60, but how this affects its function remains unclear. Here we confirm that LIP5 binds tightly to CHMP5, but also find that it binds well to additional ESCRT-III proteins including CHMP1B, CHMP2A/hVps2-1, and CHMP3/hVps24 but not CHMP4A/hSnf7-1 or CHMP6/hVps20. LIP5 binds to a different region within CHMP5 than within the other ESCRT-III proteins. In CHMP1B and CHMP2A, its binding site encompasses sequences at the proteins' extreme C-termini that overlap with "MIT interacting motifs" (MIMs) known to bind to VPS4. We find unexpected evidence of a second conserved binding site for VPS4 in CHMP2A and CHMP1B, suggesting that LIP5 and VPS4 may bind simultaneously to these proteins despite the overlap in their primary binding sites. Finally, LIP5 binds preferentially to soluble CHMP5 but instead to polymerized CHMP2A, suggesting that the newly defined interactions between LIP5 and ESCRT-III proteins may be regulated by ESCRT-III conformation. These studies point to a role for direct binding between LIP5 and ESCRT-III proteins that is likely to complement LIP5's previously described ability to regulate VPS4 activity. PMID- 18385516 TI - The role of GRASP55 in Golgi fragmentation and entry of cells into mitosis. AB - GRASP55 is a Golgi-associated protein, but its function at the Golgi remains unclear. Addition of full-length GRASP55, GRASP55-specific peptides, or an anti GRASP55 antibody inhibited Golgi fragmentation by mitotic extracts in vitro, and entry of cells into mitosis. Phospho-peptide mapping of full-length GRASP55 revealed that threonine 225 and 249 were mitotically phosphorylated. Wild-type peptides containing T225 and T249 inhibited Golgi fragmentation and entry of cells into mitosis. Mutant peptides containing T225E and T249E, in contrast, did not affect Golgi fragmentation and entry into mitosis. These findings reveal a role of GRASP55 in events leading to Golgi fragmentation and the subsequent entry of cell into mitosis. Surprisingly, however, under our experimental conditions, >85% knockdown of GRASP55 did not affect the overall organization of Golgi organization in terms of cisternal stacking and lateral connections between stacks. Based on our findings we suggest that phosphorylation of GRASP55 at T225/T249 releases a bound component, which is phosphorylated and necessary for Golgi fragmentation. Thus, GRASP55 has no role in the organization of Golgi membranes per se, but it controls their fragmentation by regulating the release of a partner, which requires a G2-specific phosphorylation at T225/T249. PMID- 18385517 TI - Cds1 controls the release of Cdc14-like phosphatase Flp1 from the nucleolus to drive full activation of the checkpoint response to replication stress in fission yeast. AB - The Cdc14p-like phosphatase Flp1p (also known as Clp1p) is regulated by cell cycle-dependent changes in its subcellular localization. Flp1p is restricted to the nucleolus and spindle pole body until prophase, when it is dispersed throughout the nucleus, mitotic spindle, and medial ring. Once released, Flp1p antagonizes Cdc2p/cyclin activity by reverting Cdc2p-phosphorylation sites on Cdc25p. On replication stress, ataxia-telangiectasia mutated/ATM/Rad3-related kinase Rad3p activates Cds1p, which phosphorylates key proteins ensuring the stability of stalled DNA replication forks. Here, we show that replication stress induces changes in the subcellular localization of Flp1p in a checkpoint dependent manner. Active Cds1p checkpoint kinase is required to release Flp1p into the nucleus. Consistently, a Flp1p mutant (flp1-9A) lacking all potential Cds1p phosphorylation sites fails to relocate in response to replication blocks and, similarly to cells lacking flp1 (Deltaflp1), presents defects in checkpoint response to replication stress. Deltaflp1 cells accumulate reduced levels of a less active Cds1p kinase in hydroxyurea (HU), indicating that nuclear Flp1p regulates Cds1p full activation. Consistently, Deltaflp1 and flp1-9A have an increased percentage of Rad22p-recombination foci during HU treatment. Together, our data show that by releasing Flp1p into the nucleus Cds1p checkpoint kinase modulates its own full activation during replication stress. PMID- 18385518 TI - Endothelin-1 couples betaPix to p66Shc: role of betaPix in cell proliferation through FOXO3a phosphorylation and p27kip1 down-regulation independently of Akt. AB - The phosphorylation of forkhead transcription factor FOXO3a by Akt is critical regulator of cell proliferation induced by serum. We show that endothelin-1 (ET 1) stimulation of primary human mesangial cells (HMCs) induces betaPix and p66Shc up-regulation, resulting in the formation of the betaPix/p66Shc complex. In transformed HMCs, ET-1 induces a biphasic phosphorylation of p66Shc and FOXO3a. The second phase leads to p27(kip1) down-regulation independently of Akt. Depletion of betaPix blocks the second phase of p66Shc and FOXO3a phosphorylation and prevents p27(kip1) down-regulation induced by ET-1. Depletion of either betaPix or p66Shc inhibits ET-1-induced cell proliferation. The expression of beta(1)Pix induces FOXO3a phosphorylation through activation of Rac1, ERK1/2, and p66Shc. Using either p66Shc- or Akt-depleted cells; we show that beta(1)Pix induced FOXO3a phosphorylation requires p66Shc but not Akt. beta(1)Pix-induced p27(kip1) down-regulation was blocked by U0126 but not by wortmannin. Endogenous betaPix and FOXO3a are constitutively associated with endogenous p66Shc. FOXO3a and p66Shc binding requires beta(1)Pix homodimerization. Expression of beta(1)Pix homodimerization deficient mutant abrogates beta(1)Pix-induced p27(kip1) down regulation and cell proliferation. Our results identify p66Shc and FOXO3a as novel partners of beta(1)Pix and represent the first direct evidence of beta(1)Pix in cell proliferation via Erk/p66Shc-dependent and Akt-independent mechanisms. PMID- 18385519 TI - Transforming growth factor-beta1-mediated Slug and Snail transcription factor up regulation reduces the density of Langerhans cells in epithelial metaplasia by affecting E-cadherin expression. AB - Epithelial metaplasia (EpM) is an acquired tissue abnormality resulting from the transformation of epithelium into another tissue with a different structure and function. This adaptative process is associated with an increased frequency of (pre)cancerous lesions. We propose that EpM is involved in cancer development by altering the expression of adhesion molecules important for cell-mediated antitumor immunity. Langerhans cells (LCs) are intraepithelial dendritic cells that initiate immune responses against viral or tumor antigens on both skin and mucosal surfaces. In the present study, we showed by immunohistology that the density of CD1a(+) LCs is reduced in EpM of the uterine cervix compared with native squamous epithelium and that the low number of LCs observed in EpM correlates with the down-regulation of cell-surface E-cadherin. We also demonstrated that transforming growth factor-beta1 is not only overexpressed in metaplastic tissues but also reduces E-cadherin expression in keratinocytes in vitro by inducing the promoter activity of Slug and Snail transcription factors. Finally, we showed that in vitro-generated LCs adhere poorly to keratinocytes transfected with either Slug or Snail DNA. These data suggest that transforming growth factor-beta1 indirectly reduces antigen-presenting cell density in EpM by affecting E-cadherin expression, which might explain the increased susceptibility of abnormal tissue differentiation to the development of cancer by the establishment of local immunodeficiency responsible for EpM tumorigenesis. PMID- 18385520 TI - Deletion of the Met tyrosine kinase in liver progenitor oval cells increases sensitivity to apoptosis in vitro. AB - The hepatocyte growth factor (HGF)/Met signaling system is essential for liver development, homeostasis, and function. In this study, we took advantage of a liver-specific, Met-conditional knockout mouse generated in our laboratory to address the molecular mechanisms of HGF/Met signaling in adult liver progenitor cell (oval cell) biology. For this purpose, we isolated oval cells from 3,5 diethoxycarbonyl-1,4-dihydro-collidine-treated Met(flx/flx) mice and established oval cell-derived cell lines that carried either functional (Met(flx/flx)) or a nonfunctional (Met(-/-)) met gene using virus-mediated Cre-loxP recombination. Oval cells lacking Met tyrosine kinase activity displayed neither Met phosphorylation nor activation of downstream targets and were refractory to HGF stimulation. Although Met(-/-) and Met(flx/flx) cells proliferated at similar rates under 10% serum, Met-deficient cells demonstrated decreased cell viability and were more prone to apoptosis when challenged with either serum starvation or the pro-apoptotic cytokine transforming growth factor-beta. Treatment with HGF reduced transforming growth factor-beta-mediated cell death in Met(flx/flx) but not Met(-/-) cells. Importantly, Met(flx/flx) and Met(-/-) cells both constitutively expressed hgf, and conditioned medium from serum-starved oval cells exhibited anti-apoptotic activity in Met(flx/flx) cells. Furthermore, serum starved Met(flx/flx) cells showed persistent activation of the Met tyrosine kinase, suggesting HGF/Met autocrine regulation. In conclusion, these data reveal a critical, functional role for Met in oval cell survival through an autocrine mechanism. PMID- 18385521 TI - Neuropeptides rescue mice from lethal sepsis by down-regulating secretion of the late-acting inflammatory mediator high mobility group box 1. AB - Originally described as a nuclear protein that bends DNA, the high mobility group box 1 protein (HMGB1) has recently emerged as a necessary and sufficient late mediator of severe sepsis. HMGB1 is therefore a molecular target that provides a wide window for clinical intervention in sepsis. Vasoactive intestinal peptide (VIP) and urocortin are two well known anti-inflammatory neuropeptides that protect against several immune disorders by regulating a wide panel of inflammatory mediators. In this study, we demonstrate the therapeutic effect of VIP and urocortin in various models of established sepsis: both agents reduced lethality induced by cecal ligation and puncture or by injection of live Escherichia coli. The therapeutic effect of VIP and urocortin was accompanied by a decrease in systemic levels of HMGB1. In addition, administration of recombinant HMGB1 completely reversed the protective effect of VIP and urocortin in experimental sepsis. In vitro and ex vivo studies show that both VIP and urocortin down-regulate translocation of HMGB1 from the nucleus to the cytoplasm and its subsequent secretion by activated macrophages, suggesting that macrophages are major targets in the inhibitory activity of these neuropeptides. To our knowledge, VIP and urocortin are the first endogenous inhibitors of HMGB1 secretion shown to improve sepsis survival in a clinically relevant time frame. PMID- 18385522 TI - Absence of alphavbeta6 integrin is linked to initiation and progression of periodontal disease. AB - Integrin alphavbeta6 is generally not expressed in adult epithelia but is induced in wound healing, cancer, and certain fibrotic disorders. Despite this generalized absence, we observed that alphavbeta6 integrin is constitutively expressed in the healthy junctional epithelium linking the gingiva to tooth enamel. Moreover, expression of alphavbeta6 integrin was down-regulated in human periodontal disease, a common medical condition causing tooth loss and also contributing to the development of cardiovascular diseases by increasing the total systemic inflammatory burden. Remarkably, integrin beta6 knockout mice developed classic signs of spontaneous, chronic periodontal disease with characteristic inflammation, epithelial down-growth, pocket formation, and bone loss around the teeth. Integrin alphavbeta6 acts as a major activator of transforming growth factor-beta1 (TGF-beta1), a key anti-inflammatory regulator in the immune system. Co-expression of TGF-beta1 and alphavbeta6 integrin was observed in the healthy junctional epithelium. Moreover, an antibody that blocks alphavbeta6 integrin-mediated activation of TGF-beta1 initiated inflammatory periodontal disease in a rat model of gingival inflammation. Thus, alphavbeta6 integrin is constitutively expressed in the epithelium sealing the gingiva to the tooth and plays a central role in protection against inflammatory periodontal disease through activation of TGF-beta1. PMID- 18385524 TI - Preservation of muscle force in Mdx3cv mice correlates with low-level expression of a near full-length dystrophin protein. AB - The complete absence of dystrophin causes Duchenne muscular dystrophy. Its restoration by greater than 20% is needed to reduce muscle pathology and improve muscle force. Dystrophin levels lower than 20% are considered therapeutically irrelevant but are associated with a less severe phenotype in certain Becker muscular dystrophy patients. To understand the role of low-level dystrophin expression, we compared muscle force and pathology in mdx3cv and mdx4cv mice. Dystrophin was eliminated in mdx4cv mouse muscle but was expressed in mdx3cv mice as a near full-length protein at approximately 5% of normal levels. Consistent with previous reports, we found dystrophic muscle pathology in both mouse strains. Surprisingly, mdx3cv extensor digitorium longus muscle showed significantly higher tetanic force and was also more resistant to eccentric contraction-induced injury than mdx4cv extensor digitorium longus muscle. Furthermore, mdx3cv mice had stronger forelimb grip strength than mdx4cv mice. Immunostaining revealed utrophin up-regulation in both mouse strains. The dystrophin-associated glycoprotein complex was also restored in the sarcolemma in both strains although at levels lower than those in normal mice. Our results suggest that subtherapeutic expression levels of near full-length, membrane-bound dystrophin, possibly in conjunction with up-regulated utrophin levels, may help maintain minimal muscle force but not arrest muscle degeneration or necrosis. Our findings provide valuable insight toward understanding delayed clinical onset and/or slow disease progression in certain Becker muscular dystrophy patients. PMID- 18385525 TI - Monte Carlo simulation of helical tomotherapy with PENELOPE. AB - Helical tomotherapy (HT) delivers intensity-modulated radiation therapy (IMRT) using the simultaneous movement of the couch, the gantry and the binary multileaf collimator (MLC), a procedure that differs from conventional dynamic or step-and shoot IMRT. A Monte Carlo (MC) simulation of HT in the helical mode therefore requires a new approach. Using validated phase-space files (PSFs) obtained through the MC simulation of the static mode with PENELOPE, an analytical model of the binary MLC, called the 'transfer function' (TF), was first devised to perform the transport of particles through the MLC much faster than time consuming MC simulation and with no significant loss of accuracy. Second, a new tool, called TomoPen, was designed to simulate the helical mode by rotating and translating the initial coordinates and directions of the particles in the PSF according to the instantaneous position of the machine, transporting the particles through the MLC (in the instantaneous configuration defined by the sinogram), and computing the dose distribution in the CT structure using PENELOPE. Good agreement with measurements and with the treatment planning system of tomotherapy was obtained, with deviations generally well within 2%/1 mm, for the simulation of the helical mode for two commissioning procedures and a clinical plan calculated and measured in homogeneous conditions. PMID- 18385523 TI - Tissue inhibitor of metalloproteinase-1 moderates airway re-epithelialization by regulating matrilysin activity. AB - Obliterative bronchiolitis (OB) is the histopathological finding in chronic lung allograft rejection. Mounting evidence suggests that epithelial damage drives the development of airway fibrosis in OB. Tissue inhibitor of metalloproteinase (TIMP)-1 expression increases in lung allografts and is associated with the onset of allograft rejection. Furthermore, in a mouse model of OB, airway obliteration is reduced in TIMP-1-deficient mice. Matrilysin (matrix metallproteinase-7) is essential for airway epithelial repair and is required for the re epithelialization of airway wounds by facilitating cell migration; therefore, the goal of this study was to determine whether TIMP-1 inhibits re-epithelialization through matrilysin. We found that TIMP-1 and matrilysin co-localized in the epithelium of human lungs with OB and both co-localized and co-immunoprecipitated in wounded primary airway epithelial cultures. TIMP-1-deficient cultures migrated faster, and epithelial cells spread to a greater extent compared with wild-type cultures. TIMP-1 also inhibited matrilysin-mediated cell migration and spreading in vitro. In vivo, TIMP-1 deficiency enhanced airway re-epithelialization after naphthalene injury. Furthermore, TIMP-1 and matrilysin co-localized in airway epithelial cells adjacent to the wound edge. Our data demonstrate that TIMP-1 interacts with matrix metalloproteinases and regulates matrilysin activity during airway epithelial repair. Furthermore, we speculate that TIMP-1 overexpression restricts airway re-epithelialization by inhibiting matrilysin activity, contributing to a stereotypic injury response that promotes airway fibrosis via bronchiole airway epithelial damage and obliteration. PMID- 18385526 TI - Thermal coagulation-induced changes of the optical properties of normal and adenomatous human colon tissues in vitro in the spectral range 400-1,100 nm. AB - The absorption coefficients, the reduced scattering coefficients and the optical penetration depths for native and coagulated human normal and adenomatous colon tissues in vitro were determined over the range of 400-1,100 nm using a spectrophotometer with an internal integrating sphere system, and the inverse adding-doubling method was applied to calculate the tissue optical properties from diffuse reflectance and total transmittance measurements. The experimental results showed that in the range of 400-1,100 nm there were larger absorption coefficients (P < 0.01) and smaller reduced scattering coefficients (P < 0.01) for adenomatous colon tissues than for normal colon tissues, and there were smaller optical penetration depths for adenomatous colon tissues than for normal colon tissues, especially in the near-infrared wavelength. Thermal coagulation induced significant increase of the absorption coefficients and reduced scattering coefficients for the normal and adenomatous colon tissues, and significantly reduced decrease of the optical penetration depths for the normal and adenomatous colon tissues. The smaller optical penetration depth for coagulated adenomatous colon tissues is a disadvantage for laser-induced thermotherapy (LITT) and photodynamic therapy (PDT). It is necessary to adjust the application parameters of lasers to achieve optimal therapy. PMID- 18385528 TI - Rathke's cleft cyst with short-term size changes in response to glucocorticoid replacement. AB - An 81-year-old man was admitted to our hospital because of general fatigue. Hormonal examination showed that he had panhypopituitarism and central diabetes insipidus. MRI imaging revealed the presence of large cystic mass with suprasellar extension in his hypothalamo-pituitary region. Interestingly, the cystic mass shrank following the start of glucocorticoid replacement, and since then relatively high doses of cortisol administration were needed to prevent the re-enlargement of cystic size. Because of the concern over possible side effects of supraphysiological doses of glucocorticoid replacement, surgical treatment was eventually carried out, confirming the pathological feature of Rathke's cleft cyst. The present case suggests that the inflammatory nature of Rathke's cleft cyst may explain the observed short-term size changes in response to glucocorticoid administration. PMID- 18385527 TI - Optimized motion estimation for MRE data with reduced motion encodes. AB - Motion estimation is an essential step common to all magnetic resonance elastography (MRE) methods. For dynamic techniques, the motion is obtained from a sinusoidal fit of the image phase at multiple, uniformly spaced relative phase offsets, phi, between the motion and the motion encoding gradients (MEGs). Generally, eight values of phi sampled at the Nyquist interval pi/4 over [0, 2pi). We introduce a method, termed reduced motion encoding (RME), that reduces the number of phi required, thereby reducing the imaging time for an MRE acquisition. A frequency-domain algorithm was implemented using the discrete Fourier transform (DFT) to derive the general least-squares solution for the motion amplitude and phase given an arbitrary number of phi. A closed form representation of the condition number of the transformation matrix which is used for estimating motion was introduced to determine the sensitivity to noise for different sampling patterns of phi. Simulation results confirmed the minimum error sampling patterns suggested from the condition number maps. The minimum noise in the motion estimate is obtained when the sampled phi are essentially evenly distributed over the range [0, pi) with an interval pi/n, where n is the number of phi sampled, or alternatively with an interval 2pi/n over the range [0, 2pi) which represents the Nyquist interval. Simulations also show that the noise level decreases as n increases as expected. The decrease in noise is the largest when n is small and it becomes less significant as n increases. The algorithm also makes it possible to estimate the motion from only two values of phi, which cannot be accomplished with traditional methods because sampling at the Nyquist interval is indeterminate. Finally, noise levels in motion estimated from phantom studies and in vivo results taken with different n agreed with that predicted by simulation and condition number calculations. PMID- 18385529 TI - Coexisting primary hyperparathyroidism and sarcoidosis in a patient with severe hypercalcemia. AB - A 75-year-old woman was admitted to our hospital because of general fatigue. She had suffered from sarcoidosis during her 40s with remission, but subsequently she experienced progression of hypercalcemia and renal dysfunction for 7 years. On admission, she showed marked hypercalcemia (up to 15.5 mg/dl) and renal failure (serum creatinine 2.5 mg/dl). Plasma intact PTH level was elevated (up to 190 pg/ml), and thyroid ultrasonography and (99m) Tc-MIBI scintigraphy detected a parathyroid mass, which was surgically removed and histologically confirmed to be a parathyroid adenoma. However, even after surgery her serum calcium remained elevated, but subsequent administration of glucocorticoid for sarcoidosis completely normalized her hypercalcemia. The simultaneous occurrence of primary hyperparathyroidism and sarcoidosis is rare, and our data suggest that high plasma PTH and 1,25(OH)D exerted an additive effect on the occurrence of severe hypercalcemia. PMID- 18385530 TI - Hypothyroidism results in small dense LDL independent of IRS traits and hypertriglyceridemia. AB - There is evidence of an association between hypothyroidism and coronary heart disease. We decided to look at the relationship between hypothyroidism and LDL subclasses' pattern including small, dense LDL to define a biochemical basis for better management of the CHD risk of these patients. We utilized a case-control design to evaluate differences in lipid parameters between cases and controls. Univariate analysis revealed that many factors were associated with LDL particle size. Binary logistic regression however revealed that only thyroid status and serum triglyceride (TG) levels were independently associated with LDL particle size. Results from this study support an independent association between LDL particle size phenotype and both plasma TG concentrations and thyroid status. After adjusting for TG levels, other insulin resistance syndrome (IRS) traits were not associated with LDL size phenotype, suggesting that the IRS related sdLDL is linked most strongly to alterations in TG levels. PMID- 18385531 TI - Clinical course of patients with nonclassical 21-hydroxylase deficiency (21-OHD) diagnosed in infancy and childhood. AB - We report four cases of nonclassical 21-hydroxylase deficiency (21-OHD) diagnosed in neonate or early childhood. The four patients comprised a 6-year, 5-month-old male (case 1); a 3-year, 10-month-old female (case 2); a 13-year, 11-month-old female (case 3) and a 17-year, 1-month-old male (case 4). Cases 3 and 4 were siblings. None had any signs of virilization or salt wasting at birth. 21-OHD was diagnosed using ACTH loading test and other adrenal steroid evaluations. Mutations of the CYP21 gene were detected in all patients. Three patients (cases 1, 3 and 4) had positive results in neonatal mass screening. Cases 1 and 2 showed no apparent signs of virilization and were observed without conventional treatment. In cases 3 and 4, because of increased growth velocity and accelerated bone maturation, hydrocortisone administration was initiated from their late infantile period. In spite of hydrocortisone treatment, in case 4, the final height of 159.7 cm was less than his predicted final height. Besides he revealed adrenal insufficiency at the age of 9 years and 2 months old caused by viral infection. Hydrocortisone supplementation therapy may cause adrenal insufficiency in nonclassical patients due to suppression of the hypothalamus-pituitary-adrenal axis. The clinical courses in these cases were various, and it was difficult to predict the appearance of any symptoms of virilization. Careful observation is necessary. PMID- 18385532 TI - Effects of DHMEQ, a novel nuclear factor-kappab inhibitor, on beta cell dysfunction in INS-1 cells. AB - AIMS: Recent studies suggest that nuclear factor-kappaB (NF-kappaB) activation has an important role in leading to beta cell dysfunction in both type 1 and type 2 diabetes. In this study we tested this hypothesis by investigating the effects of dehydroxymethylepoxyquinomicin (DHMEQ), a novel NF-kappaB inhibitor, on tumor necrosis factor-alpha (TNF-alpha)-induced beta cell dysfunction. METHODS: INS-1 cells were incubated with TNF- alpha and with or without DHMEQ for 24 hours. Glucose-stimulated insulin secretion, cell viability, mRNA expression and NF kappaB activation were investigated. RESULTS: DHMEQ suppressed TNF-alpha-induced NF-kappaB activation and partially ameliorated glucose-stimulated insulin secretion in a dose-dependent manner. DHMEQ also partially ameliorated decreased cell viability and insulin mRNA level induced by TNF-alpha. CONCLUSION: DHMEQ suppressed NF-kappaB activation and ameliorated beta cell dysfunction induced by TNF- alpha. Inhibition of activated NF-kappaB in beta cells may be important to ameliorate beta cell dysfunction in diabetes. PMID- 18385533 TI - Antiatherogenic functionality of high density lipoprotein: how much versus how good. AB - Plasma concentration of high density lipoprotein (HDL) is one of the most reliable negative risk factors for CVD. There is however convincing experimental and clinical evidence that plasma concentration of HDL does not convey the full picture of atheroprotective properties of HDL. HDL functionality, i.e. the ability of HDL to perform its many atheroprotective functions, is partly independent of HDL concentration and may be as important, if not more important, in determining the atheroprotective capacity of HDL. The capacity of HDL to support cholesterol efflux, its anti-inflammatory, anti-oxidant, anti-thrombotic and other atheroprotective functions are affected dramatically in conditions like coronary artery disease, chronic and acute inflammation, diabetes as well as through various interventions. The mechanisms connecting changes in HDL functionality to HDL structure are only beginning to emerge. Modifications of HDL proteins and lipids, such as advanced glycation and oxidation, changes in HDL composition and size of HDL particles, changes in abundance of various proteins and lipids carried by HDL are among factors affecting HDL functionality. A single common denominator reflecting the multiple HDL functions is yet to be found and may not exist leaving direct measurements of each HDL function as the way to assess atheroprotective capacity of HDL. PMID- 18385534 TI - Mechanism of pi-induced vascular calcification. AB - Vascular calcification is clinically important in the development of cardiovascular disease. It has been suggested that apoptosis is one of the processes regulating calcification in vascular smooth muscle cells (VSMC). In this review, we discuss the role of apoptosis in inorganic phosphate (Pi)- induced calcification, focusing on regulation of the survival pathway mediated by growth arrest- specific gene 6 (Gas6). Further, we mention the beneficial effect of statins mediated by inhibition of apoptosis which is accompanied by restoration of the Gas6-mediated survival pathway. These findings indicate that Gas6 is a novel target of statins' effects to prevent vascular calcification. PMID- 18385535 TI - Revised optimal cut-off point of waist circumference for the diagnosis of metabolic syndrome in Japanese women and the influence of height. AB - AIM: We reevaluated waist circumference as a diagnostic criterion of metabolic syndrome (MetS) in Japanese. METHODS: We enrolled 5,571 subjects (3,148 men and 2,423 women) who had health check-ups in our center. The criterion was reevaluated using the positive predictive value of a receiver-operating characteristics (ROC) curve at 10 different hypothesized lengths of waist circumference with or without a cluster of risk factors. We also drew ROC curves based on the atherosclerotic findings of clinical examinations. RESULTS: Based on the ROC curves, the optimal waist circumference cut-off was 85 cm in men and 80 cm in women. Using this 80 cm cut-off point in women, misdiagnosis rates of MetS were lowered (-19.1--56.6%) compared to the cut-off point currently in use. Integrating the influence of height, namely by using a waist-to-height(2) ratio, misdiagnosis rates in shorter populations were decreased in both men and women. CONCLUSION: These data suggested an optimal waist circumference cut-off to improve the diagnostic probability of MetS in Japanese women of 80 cm, as well as the utility of an easily detected anthropometric index such as a waist-to-height (cm x 100/cm) or waist-to-height(2) (cm x 10,000/cm(2)) ratio, determined as 51 in men and 52 in women, or 30 in men and 33 in women, respectively. PMID- 18385536 TI - Pitavastatin improves serum resistin levels in patients with hypercholesterolemia. AB - AIM: Resistin in serum is associated with high risk in patients with atherosclerosis. This clinical study aimed to investigate whether pitavastatin can regulate the serum level of resistin, together with levels of other inflammatory cytokines and adipocytokines. METHODS: Forty two outpatients (mean age 65.2 +/- 12.6 yr, M/F: 21/21) with hypercholesterolemia were administered 2 mg of pitavastatin and serum levels of resistin, together with serum levels of adiponectin, leptin, TNF-alpha and hsCRP, were measured before, and 12 weeks after enrollment. RESULTS: There was no significant gender-related difference in initial serum resistin levels. Pitavastatin significantly decreased LDL cholesterol after 12 weeks. Initial levels of resistin showed a significant correlation with those of hsCRP (r=0.38, p=0.013), but not TNF-alpha or HOMA-R. Serum resistin, but not adiponectin and leptin, levels were significantly decreased, dropping from 17.1 +/- 9.9 ng/ dL to 15.2+/-10.0 (p=0.001) after 12 weeks of administration. The patient group with a baseline hsCRP > or = 0.1 at enrollment (n=17) had decreased levels of both resistin and hsCRP (p=0.011 and p=0.022, respectively). CONCLUSION: This study showed the pleiotropic effect of pitavastatin on the serum resistin concentration, suggesting that it may assist in the prevention of atherosclerosis. PMID- 18385537 TI - Evaluation of a homogeneous assay for measuring LDL-cholesterol in hyperlipidemic serum specimens. AB - AIM: Homogeneous assay reagents for the determination of low-density lipoprotein cholesterol (LDL-C) have been available from several manufacturers. However, there has been considerable controversy due to uncertainty regarding their reactivity with intermediate-density lipoprotein (IDL), which is detected at an especially high frequency in patients with type III hyperlipemia. In this study, we examined the reactivity of a homogeneous assay, Cholestest LDL (R) (CT-LDL), with hyperlipemic sera that were classified according to the WHO system. METHODS: Sera from 6 normolipidemic and 22 hyperlipidemic patients classified according to the WHO system were used for this study. All serum specimens were fractionated by the ultracentrifugation method of Hatch and Lees, and subjected to lipid and protein measurements. RESULTS: The percent bias of values measured by CT-LDL relative to those determined by the ultracentrifugation method was calculated and compared to the lipid/protein ratios of each lipoprotein fraction. Consequently, the coefficient of correlation between the bias and the Triglyceride/Total cholesterol (TG/TC) ratio in the IDL fraction was 0.742. There were also correlations with the TG/TC ratio and the apo-lipoprotein B/Total Cholesterol ratio in the LDL fraction and in the LDL+IDL fraction, respectively. CONCLUSION: Further testing will be required in order to know more about the clinical condition of hyperlipidemic patients, since CT-LDL may react differently with some beta lipoproteins having a diverse lipid/protein composition compared to those in normolipidemic specimens. PMID- 18385538 TI - Jun N-terminal kinase inhibitor blocks angiogenesis by blocking VEGF secretion and an MMP pathway. AB - AIM: The excessive proliferation and migration of vascular smooth muscle cells (SMCs) and angiogenesis of endothelial cells (ECs) participate in the growth and instability of atherosclerotic plaques. It is unclear whether Jun N-terminal kinase (JNK) is pro-or anti-atherogenic. METHODS: We examined the direct effect of JNK inhibitor (JNK-I) on the proliferation and formation of tubes by human coronary SMCs and human coronary ECs. RESULTS: Culture medium from JNK-I-treated SMCs prevented ECs from forming tubes in an in vitro model of angiogenesis indirectly by reducing the amount of vascular endothelial growth factor (VEGF) released from SMCs. In addition, JNK-I attenuated the expression of pro-matrix metalloproteinase-2 in ECs. When added back to the medium of SMCs treated with JNK-I, VEGF blocked the inhibitory effect on the formation of tubes. CONCLUSION: Our results indicate JNK-I to have a direct anti-atherogenic effect in SMCs and ECs. PMID- 18385539 TI - Systemic glucocorticoid therapy reduces pain and the number of endoneurial tumor necrosis factor-alpha (TNFalpha)-positive mast cells in rats with a painful peripheral neuropathy. AB - Clinical and experimental evidence suggests that glucocorticoids may be effective in the treatment of neuropathic pain, but their mechanism of action is unknown. We gave triamcinolone (3 mg/kg) to rats with an experimental post-traumatic painful peripheral neuropathy, chronic constriction injury (CCI), five days after nerve injury, when the abnormal pain syndrome is known to be present; and pain sensitivity was measured on postoperative days 7 - 14, a period during which symptoms are known to be at approximately peak severity. Additional CCI rats were treated similarly; and then they were sacrificed five days after the injection for an immunocytochemical analysis of endoneurial tumor necrosis factor-alpha (TNFalpha), macrophages, and mast cells in the sciatic nerve proximal to the site of injury. Vehicle-injected CCI rats demonstrated the expected neuropathic pain symptoms. Triamcinolone-treated CCI rats had a statistically significant reduction in the magnitude of heat-hyperalgesia and mechano-allodynia, but there was no effect on cold-allodynia or mechano-hyperalgesia. On the nerve-injured side of vehicle-injected rats, TNFalpha was present in Schwann cells and mast cells. On the nerve-injured side of triamcinolone-treated rats, there was a significant (71.5%) reduction in the number of TNFalpha-positive mast cells. Our results suggest that glucocorticoid therapy for neuropathic pain may work via the reduced expression of TNFalpha in endoneurial mast cells. PMID- 18385540 TI - Comparison of the protective effect of N-acetylcysteine by different treatments on rat myocardial ischemia-reperfusion injury. AB - Reactive oxygen species have been known as important contributors to ischemia/reperfusion (I/R) injury. Studies on the beneficial effect of N acetylcysteine (NAC), a potent antioxidant, on limiting infarct size induced by I/R yielded contrasting results. The present study was undertaken to compare the effect of NAC by different administration methods on infarct size in a rat myocardial I/R model. Rats underwent 30 min of left coronary occlusion followed by 4 h of reperfusion. Treatment with continuous infusion of NAC (150 mg/kg per hour) from 30 min before occlusion for 2 h (until 1 h after the start of reperfusion) produced a significant limitation of the infarct size as a percentage of the ischemic area (8%) compared to the non-treated control (60%). However, bolus injection of 150 mg/kg at 30 min prior to occlusion and 5 min prior to reperfusion failed to reduce it (56%) although the total dose is the same. The decreased total glutathione content and glutathione peroxidase activity in the ischemic region were recovered in the continuous infusion group, but not in the bolus injection group. The increased myeloperoxidase activity and phosphorylation of inhibitor kappaB after I/R were inhibited by the continuous treatment. These results indicate that the protective effect of NAC on myocardial infarction induced by I/R was different depending on the administration method. It is necessary to maintain blood concentration during the early period of reperfusion to obtain the beneficial effect of NAC. PMID- 18385541 TI - Vulnerability of gastric mucosa to prednisolone in rats chronically exposed to cigarette smoke. AB - We examined gastric mucosal vulnerability in a rat model of chronic obstructive pulmonary disease (COPD). Male Wistar rats were exposed to cigarette smoke for 12 weeks (CSE rats), and on the last 4 days of exposure, prednisolone was given to induce gastric mucosal injury. Histopathology, pulmonary function, arterial blood gases, and levels of lipid peroxides (LPO), prostaglandin E(2) (PGE(2)), hypoxia inducible factor 1 alpha subunit (HIF-1alpha), and vascular endothelial growth factor (VEGF) in gastric mucosa were examined. We also tested the effect of rebamipide on prednisolone-induced gastric lesions. In CSE rats, although no gastric lesions were detected, LPO, PGE(2), HIF-1alpha, and VEGF levels were higher than in control rats. Prednisolone induced gastric hemorrhagic lesions more readily in CSE rats than controls, with concomitant decrease in PaO(2) and increased levels of LPO, HIF-1alpha, and VEGF. Rebamipide reversed gastric lesions without affecting any parameters examined. CSE rats were found to be a useful animal model of COPD, and COPD appeared to render the gastric mucosa vulnerable to prednisolone. PMID- 18385542 TI - GABAC-receptor stimulation activates cAMP-dependent protein kinase via A-kinase anchoring protein 220. AB - In our previous study, anti-apoptotic effects of GABA(C)-receptor stimulation was suppressed by inhibitors of cAMP-dependent protein kinase (PKA), implying GABA(C) receptor-mediated PKA activation. The present study showed that GABA(C)-receptor stimulation with its agonist, cis-4-aminocrotonic acid (CACA), protected cultured hippocampal neurons from amyloid beta 25 - 35 (Abeta25 - 35) peptide-enhanced glutamate neurotoxicity. This protective effect of CACA was blocked by PKA inhibitors, KT 5720 and H-89, as well as a specific GABA(C)-receptor antagonist, (1,2,5,6-tetrahydropyridine-4-yl) methylphosphinic acid (TPMPA). To test the possibility of GABA(C) receptor-mediated PKA activation, association of GABA(C) receptor with A-kinase anchoring proteins (AKAPs) and effect of an AKAP antisense oligonucleotide on the PKA activation were examined in primary cultured rat hippocampal neurons. Stimulation of the cells with CACA-activated PKA was assessed by the phosphorylated PKA substrate (135 kDa) level. Specific antibodies raised against GABA(C)-receptor rho subunits precipitated each rho subunit, AKAP220, and PKA regulatory and catalytic subunits from rat brain lysates, suggesting that rho is associated with the AKAP220/PKA complex. Furthermore, antisense oligonucleotide of AKAP220 suppressed such GABA(C) stimulation-induced PKA activation, suggesting that GABA(C)-receptor stimulation activates PKA via AKAP220. PMID- 18385543 TI - Leydig cell hyperplasia in an ENU-induced mutant mouse with germ cell depletion. AB - Repro22 is an N-ethyl-N-nitrosourea (ENU)-induced mutation in mice showing depletion of both male and female germ cells. In the present study, we investigated the male phenotypes of the mutant mouse at the adult stage. The repro22/repro22 homozygous mice showed reduced body weights as well as markedly reduced testis weights. Histological examination of the testes at 4 and 10 months of age showed no germ cells in the seminiferous tubules of the affected testis while a number of Sertoli cells were observed in the tubules. In addition to the germ cell depletion, the testes of the affected mouse contained expanded intertubular spaces that were filled by Leydig cell-like interstitial cells. These interstitial cells were confirmed to be Leydig cells by immunohistochmical staining using anti-3beta-HSD antibody. The estimated number of Leydig cells in the affected testes at 10 months of age increased approximately 2 fold compared with those of normal testes. Furthermore, the plasma testosterone levels of the affected mice at 10 months of age were significantly higher than those of the normal mice. These findings indicated that the repro22/repro22 mouse developed hyperplasia of Leydig cells that was presumably caused by the absence of germ cells in the seminiferous tubules. PMID- 18385545 TI - [Use of economic evaluation in the listing and pricing of pharmaceuticals]. AB - To curb a rapid increase in expenditures for pharmaceuticals, the Korean government introduced a positive list system and a negotiation process for drug prices at the end of 2006. Economic evaluation of pharmaceuticals has begun to have a pivotal role in the listing and pricing of drugs for the Korean National Health Insurance. There are some points to discuss regarding the use of economic evaluation in the listing and pricing in the context of the Korean system. First, the listing and pricing processes have been fragmented, evoking complaints from pharmaceutical companies and delaying the access of new drugs to patients. Second, there is a concern that the positive list system may limit the range and availability of drugs for patients to choose for treatment. Third, the time schedule for de-listing of existing drugs may not be realistic. Fourth, it is not always easy to provide reliable evidence of cost-effectiveness due to a lack of materials. Fifth, there is no consensus on the range of the ICER (incremental cost-effectiveness ratio) acceptable to the Korean society. In conclusion, in the near future, it will be necessary to evaluate the achievements that the economic evaluation has provided to the Korean society. PMID- 18385546 TI - [Current state and challenges of pharmacoeconomic evaluation in Korea]. AB - Since the positive listing system for prescription drug reimbursement has been introduced in Korea, the number of pharmacoeconomic evaluation studies has increased. However it is not clear if the quality of pharmacoeconomic evaluation study has improved. Due to the lack of randomized clinical studies in Korean health care setting, Korean economic evaluation studies have typically integrated the local cost data and foreign clinical data. Therefore methodological issues can be raised in regard to data coherence and consistency. But the quality of data was not questiened and the potential bias has not been investigated yet. Even though changes in policy have encouraged the undertaking of pharmacoeconomic evaluations, there is few public-side funding for validation study of cost effectiveness models and data. Several companies perform economic evaluation studies to be submitted on behalf of their own products, but do not want the study results to be disclosed to the academic community or public. To improve the present conduct of pharmacoeconomic evaluations in Korea, various funding sources need to be developed, and, like other multidisciplinary areas, the experts in different fields of study should collaborate to ensure the validity and credibility of pharmacoeconomic evaluations. PMID- 18385547 TI - [Guidelines for economic evaluation of pharmaceuticals in Korea]. AB - An economic evaluation is required in order to apply to the Health Insurance Review and Assessment Service (HIRA) for a listing in the national drug formulary. To assist companies in preparing the necessary documents, HIRA published guidelines for the economic evaluation of pharmaceuticals in 2006. The guidelines are composed of two parts: guidance an explanatory notes. Each guideline reflects the best practice which meets both the theoretical consensus within the academic community and local situations, like data availability. To enhance the transparency of evaluation, guidelines emphasize the reproducibility of data and analysis result. That is, all evaluation processes are required to be described in enough detail to be replicated by reviewers. With growing experience and theoretical development in this area, HIRA guidelines will be revised periodically. PMID- 18385548 TI - [Global activity of cancer registries and cancer control and cancer incidence statistics in Korea]. AB - The World Health Organization (WHO) reported that cancer killed 7.6 million people in the world in 2005, and that 40% of all cancer deaths can be prevented. According to the WHO Global Action Plan Against Cancer (GAPAC), monitoring of cancer patients is the essential part of cancer control, and should be conducted through cancer registration. Originally, cancer registries were primarily concerned with the description of cancer patterns, trends of cancer occurrence, and etiology of cancer. In the last 20 years, cancer registries provided not only information on the incidence and characteristics of specific cancers, but also supplied the source of cancer control planning and evaluation and the care of individual cancer patients with survival. Cancer Incidence in Five Continents (CI5) presents incidence data from populations all over the world every five year. Volume IX in the series (data for 1998--2002) has recently (November 2007) been published online at International Agency for Research on Cancer (IARC). Nine data from Korea Central Cancer Registry (National data), Seoul, Busan, Daegu, Gwangju, Incheon, Daejeon, Usan, Jejudo regional cancer registries were included in that volume. In this paper, the editorial process, the characteristics of national data, and quality indices in CI5 IX are being described. In addition, cancer control activities related to cancer registration in some selected countries are also presented. PMID- 18385549 TI - [Ten year trend of cancer incidence in Seoul, Korea: 1993--2002]. AB - OBJECTIVES: Effective cancer prevention and control measures can only be done when dependable data on the cancer incidence is available. The Seoul Cancer Registry (SCR) was founded to provide valid, comparable and representative cancer incidence data for Koreans. We aimed to compare the cancer incidence in the first (1993--1997) and second term (1998--2002) of the SCR, and we analyzed the annual incidence trend during that 10 years. METHODS: The SCR detects potential cancer cases through the Korean Central Cancer Registry (KCCR) data, the health insurance claims, the individual hospital's discharge records and the death certificates. About 87% of the SCR data is registered through the KCCR. The rest of the data is registered by SCR registrars who visit about 70-80 mid-sized hospitals in Seoul to review and abstract the medical records of the potential cancer patients. RESULTS: The total number of new cancer cases was higher in 1998 -2002 than in 1993--1997 by 20.6% for men and 18.4% for women, respectively. The age-standardized rate (ASR) of total cancer per 100,000 increased 1% (from 295.4 to 298.3) for men and 5.1% (from 181.5 to 190.7) for women, between the two periods. The commonest cancer sites during 1998--2002 for men were stomach, liver, bronchus/lung, colorectum, bladder and prostate, and the commonest cancer sites for women were breast, stomach, colorectum, cervix uteri, thyroid and bronchus/lung. Compared with the ASRs in 1993, the ASRs in 2002 increased for colorectum (58.4% for men, 27.1% for women), prostate (81.5%), breast (58.3% for women), thyroid (141% for women), and bronchus/lung (15.4% for women). The ASRs for stomach (-18.7% for men, -20.7% for women) and uterine cervix cancer (-39.7%) had decreased. CONCLUSIONS: The cancer incidence is increasing in Seoul, Korea, especially for the colorectum and prostate for men, and for the breast, colorectum, bronchus/lung and thyroid for women. PMID- 18385550 TI - [Factors associated with gastric cancer screening of Koreans based on a socio ecological model]. AB - OBJECTIVES: We measured behavioral factors associated with Koreans receiving gastric cancer screening based on a socio-ecological model, in part to develop strategies to improve cancer screening rates. METHODS: A telephone survey was conducted with 2,576 people chosen through stratified random sampling from April 1--May 31, 2004. Collected information included gastric cancer screening, socio demographic factors, and socio-ecological factors at intrapersonal, interpersonal, community, and public policy levels. RESULTS: Among 985 survey respondents (380 men and 605 women), 402 had received gastric cancer screening. Logistic analysis was performed to compare those screened and unscreened. 'Age' was the only demographic factor that showed a statistically significant association with getting screening. People in their fifties (OR=1.731, 95% CI=1.190-2.520) and sixties (OR=2.098, 95% CI=1.301-3.385) showed a higher likelihood of getting screened, compared to those in the forties. 'Accessibility to a medical institution' was a significant factor related to having gastric cancer screening at the intrapersonal level. At the interpersonal level, recommendations by family members to be screened and a family practice of routine cancer screening were significantly related. People with frequent education about cancer screening or with stronger social feelings that cancer screening is necessary also demonstrated significantly higher tendencies to be screened. CONCLUSIONS: In conclusion, a socio-ecological model seems appropriate for explaining gastric cancer screening behavior and associated factors. Health planners should develop integrated strategies to improve cancer screening rates based on socio-ecological factors, especially at the interpersonal and community levels. PMID- 18385551 TI - [The association between obesity indices in adolescence and carotid intima-media thickness in young adults: Kangwha Study]. AB - OBJECTIVES: The aim of this study is to investigate the association between obesity indices (body mass index, weight, waist-hip ratio and waist circumference) in adolescents and the carotid intima-media thickness (C-IMT) in early adulthood. We also wanted to identify the best predictor for C-IMT among these obesity indices. METHODS: This study used community-based prospective cohort study, known as the Kangwha Study, and the data we used were from subjects who were 16-years old in 1996 (defined as "adolescence") and 25 years-old in 2005 (defined as "early adulthood"). The 256 subjects (113 men and 143 women) who were used for analysis participated in both follow-ups, and they underwent B-mode ultrasonography of the carotid arteries at the early adulthood follow-up. Obesity indices were defined as the body mass index, weight, waist-hip ratio and waist circumference. The C-IMT was defined as the mean of the maximal IMT of each common carotid artery. The C-IMT and obesity indices associations were evaluated via multivariable regression, logistic regression and the receiver-operator characteristic curve analyses. RESULTS: In men, all the obesity indices in adolescence were showed to have statistically significant positive association with C-IMT in early adulthood. However, no such relationship was showed in women. On multiple regression and logistic regression analysis, the waist-hip ratio showed the biggest relationship with the C-IMT among the 4 obesity indices. However, there were no statistical significant differences and no best predictor was found. For the women, the obesity incidences and C-IMT showed no relationships. CONCLUSIONS: This study suggested that obesity in adolescence was related to an increase C-IMT in healthy young Korean men. PMID- 18385552 TI - [Serum ferritin and risk of the metabolic syndrome in some Korean rural residents]. AB - OBJECTIVES: The purpose of this study was to examine the relationship between serum ferritin and the metabolic syndrome (MS). METHODS: We conducted a cross sectional study of 1,444 adults over age 40 and under age 70 that lived in a rural area and participated in a survey conducted as part of the Korean Rural Genomic Cohort Study (KRGCS). The MS was defined as the presence of at least three of the followings: elevated blood pressure, low high density lipoprotein cholesterol, elevated serum triglycerides, elevated plasma glucose, or abdominal obesity. After adjustment for age, alcohol intake, menopausal status, body mass index (BMI), high sensitivity C-reactive protein (hs-CRP), and alanine aminotransferase (ALT), odds ratios (ORs) for the prevalence of the MS by sex were calculated for quartiles of serum ferritin using logistic regression analysis. RESULTS: The MS was more common in those persons with the highest levels of serum ferritin, compared to persons with the lowest levels, in men (37.1% vs. 22.4%, p=0.006) and women (58.8% vs. 34.8, p<0.001). In both sexes, the greater the number of MS components presents, the greater the serum ferritin levels. After adjustment for age, alcohol intake, and menopausal status, the OR for metabolic syndrome, comparing the fourth quartile of ferritin with the first quartile, was 2.21 (95% confidence interval; CI=1.26-3.87; p-trend=0.024) in men and 2.10 (95% CI=1.40-3.17; p-trend=0.001) in women. However, after further adjustment for BMI, hs-CRP, and ALT, the ORs were statistically attenuated in both sexes. CONCLUSIONS: Moderately elevated serum ferritin levels were not independently associated with the prevalence of the MS after adjusting for other risk factors. Further studies are needed to obtain evidence concerning the association between serum ferritin levels and the MS. PMID- 18385553 TI - [The analysis of health related behavior after using health information on the Internet]. AB - OBJECTIVES: This study investigated the health information such as the general health information, the health product information, and the hospital information, and we wanted to identify the association between internet health information and the health related behavior by analyzing the process after people search the Internet. METHODS: A telephone survey with structured questionnaire was performed by trained surveyors. The respondents were sampled proportionate to the Korean demographic distribution with considering the city size and the populations' ages and gender. The survey was conducted from October 2006 to November 2006. RESULTS: Out of 3,758 successfully connected persons of age 20 or more, 871(23.2%) respondents had used Internet health information during the last year. The purposes of searching the Internet for health was, 1) to get general health information (717 cases, 81.0%), 2) shopping for health product (109 cases, 12.3%) and 3) seeking information about hospital selection (59 cases, 6.7%). Our research showed that the process after searching the Internet for health information depends on the purpose of the search. 68.8% of the searchers for general health information, 67% of the searchers for health product shopping and 64.4% of the searchers seeking information to guide hospital selection were satisfied with their Internet search. However one third of the respondents reported not being satisfied with the result of the search. CONCLUSIONS: Unsatisfied consumers with internet health information tended to ask lay referrals from others or they gave up seeking health information. The health information system should be improved to increase the accessibility and to provide reliable and effective information. Also, a more user-centric community is needed in order to strengthen the effective role of lay referrals among the internet users. PMID- 18385554 TI - [Reference interval of serum thyroid hormones in healthy Korean adults]. AB - OBJECTIVES: This study was conducted to determine the reference interval of serum thyroid hormones (TSH, FT3, FT4) in healthy Korean adults. METHODS: Health examination data from 1,591 healthy Korean adults who visited an university hospital were analyzed. Patients with specific health conditions capable of altering laboratory results were excluded from the study. Serum thyroid hormones were measured using IMMULITE 2000 (DPC, USA, 2002). Subjects were 18-65 years old; 911 were male, and 690 were female. RESULTS: The arithmetic means of TSH, FT3, and FT4 values for male subjects were 1.28+/-1.84 microIU/ml, 3.23+/-0.57 pg/ml, and 1.42+/-0.22 ng/dl, respectively. In female subjects, the arithmetic means of TSH, FT3, and FT4 values were 1.49+/-2.08 microIU/ml, 3.08+/-0.54 pg/ml, and 1.29 +/-0.24 ng/dl, respectively. The arithmetic mean FT4 value for males decreased with age (p<0.01). The arithmetic mean FT3 value for females increased with age (p<0.01). The arithmetic mean thyroid hormone values of all study subjects differed significantly based on season. The arithmetic mean of male FT4 decreased with increasing BMI (p<0.01). The arithmetic mean of female FT3 increased with increasing BMI (p<0.01). The reference intervals recommended by the IMMULITE 2000 manufacturer are 0.40-4.00 microIU/ml for TSH, 1.80-4.20 pg/ml for FT3, and 0.80-1.90 ng/dl for FT4 (same values for both genders). CONCLUSIONS: There was a significant difference in the interval of thyroid hormones between males and females, but the reference interval of IMMULITE 2000 was not established by gender. There is a need to reestablish the reference interval for thyroid hormones in Korean healthy adults. PMID- 18385555 TI - Ultra-low-dose MDCT of the chest: influence on automated lung nodule detection. AB - OBJECTIVE: To evaluate the relationship between CT dose and the performance of a computer-aided diagnosis (CAD) system, and to determine how best to minimize patient exposure to ionizing radiation while maintaining sufficient image quality for automated lung nodule detection, by the use of lung cancer screening CT. MATERIALS AND METHODS: Twenty-five asymptomatic volunteers participated in the study. Each volunteer underwent a low-dose CT scan without contrast enhancement (multidetector CT with 16 detector rows, 1.25 mm section thickness, 120 kVp, beam pitch 1.35, 0.6 second rotation time, with 1.25 mm thickness reconstruction at 1.25 mm intervals) using four different amperages 32, 16, 8, and 4 mAs. All series were analyzed using a commercially available CAD system for automatic lung nodule detection and the results were reviewed by a consensus reading by two radiologists. The McNemar test and Kappa analysis were used to compare differences in terms of the abilities to detect pulmonary nodules. RESULTS: A total of 78 non-calcified true nodules were visualized in the 25 study subjects. The sensitivities for nodule detection were as follows: 72% at 32 mAs, 64% at 16 mAs, 59% at 8 mAs, and 40% at 4 mAs. Although the overall nodule-detecting performance was best at 32 mAs, no significant difference in nodule detectability was observed between scans at 16 mAs or 8 mAs versus 32 mAs. However, scans performed at 4 mAs were significantly inferior to those performed at 32 mAs (p < 0.001). CONCLUSION: Reducing the radiation dose (i.e. reducing the amperage) lowers lung nodule detectability by CAD. However, relatively low dose scans were found to be acceptable and to cause no significant reduction in nodule detectability versus usual low-dose CT. PMID- 18385556 TI - Estimating the absorbed dose to critical organs during dual X-ray absorptiometry. AB - OBJECTIVE: The purpose of this study is to estimate a patient's organ dose (effective dose) during performance of dual X-ray absorptiometry by using the correlations derived from the surface dose and the depth doses in an anthropomorphic phantom. MATERIALS AND METHODS: An anthropomorphic phantom was designed and TLDs (Thermoluminescent Dosimeters) were placed at the surface and these were also inserted at different depths of the thyroid and uterus of the anthropomorphic phantom. The absorbed doses were measured on the phantom for the spine and femur scan modes. The correlation coefficients and regression functions between the absorbed surface dose and the depth dose were determined. The derived correlation was then applied for 40 women patients to estimate the depth doses to the thyroid and uterus. RESULTS: There was a correlation between the surface dose and depth dose of the thyroid and uterus in both scan modes. For the women's dosimetry, the average surface doses of the thyroid and uterus were 1.88 microGy and 1.81 microGy, respectively. Also, the scan center dose in the women was 5.70 microGy. There was correlation between the thyroid and uterus surface doses, and the scan center dose. CONCLUSION: We concluded that the effective dose to the patient's critical organs during dual X-ray absorptiometry can be estimated by the correlation derived from phantom dosimetry. PMID- 18385557 TI - Nonpalpable breast masses: evaluation by US elastography. AB - OBJECTIVE: To compare the diagnostic performances of conventional ultrasound (US) and US elastography for the differentiation of nonpalpable breast masses, and to evaluate whether elastography is helpful at reducing the number of benign biopsies, using histological analysis as a reference standard. MATERIALS AND METHODS: Conventional US and real-time elastographic images were obtained for 100 women who had been scheduled for a US-guided core biopsy of 100 nonpalpable breast masses (83 benign, 17 malignant). Two experienced radiologists unaware of the biopsy and clinical findings analyzed conventional US and elastographic images by consensus, and classified lesions based on degree of suspicion regarding the probability of malignancy. Results were evaluated by receiver operating characteristic curve analysis. In addition, the authors investigated whether a subset of lesions was categorized as suspicious by conventional US, but as benign by elastography. RESULTS: Areas under the ROC curves (Az values) were 0.901 for conventional US and 0.916 for elastography (p = 0.808). For BI-RADS category 4a lesions, 44% (22 of 50) had an elasticity score of 1 and all were found to be benign. CONCLUSION: Elastography was found to have a diagnostic performance comparable to that of conventional US for the differentiation of nonpalpable breast masses. The authors conclude that BI-RADS category 4a lesions with an elasticity score of 1 probably do not require biopsy. PMID- 18385558 TI - Advanced gastric cancer and perfusion imaging using a multidetector row computed tomography: correlation with prognostic determinants. AB - OBJECTIVE: To investigate the relationship between the perfusion CT features and the clinicopathologically determined prognostic factors in advanced gastric cancer cases. MATERIALS AND METHODS: A perfusion CT was performed on 31 patients with gastric cancer one week before surgery using a 16-channel multi-detector CT (MDCT) instrument. The data were analyzed with commercially available software to calculate tumor blood flow (BF), blood volume (BV), mean transit time (MTT), and permeability surface (PS). The microvessel density (MVD), was evaluated by immunohistochemical staining of the surgical specimens with anti-CD34. All of the findings were analyzed prospectively and correlated with the clinicopathological findings, which included histological grading, presence of lymph node metastasis, serosal involvement, distant metastasis, tumor, node, metastasis (TNM) staging, and MVD. The statistical analyses used included the Student's t-test and the Spearman rank correlation were performed in SPSS 11.5. RESULTS: The mean perfusion values and MVD for tumors were as follows: BF (48.14+/-16.46 ml/100 g/min), BV (6.70+/-2.95 ml/100 g), MTT (11.75+/-4.02 s), PS (14.17+/-5.23 ml/100 g/min) and MVD (41.7+/-11.53). Moreover, a significant difference in the PS values was found between patients with or without lymphatic involvement (p = 0.038), as well as with different histological grades (p = 0.04) and TNM stagings (p = 0.026). However, BF, BV, MTT, and MVD of gastric cancer revealed no significant relationship with the clinicopathological findings described above (p > 0.05). CONCLUSION: The perfusion CT values of the permeable surface could serve as a useful prognostic indicator in patients with advanced gastric cancer. PMID- 18385559 TI - Power Doppler imaging in acute renal vein occlusion and recanalization: a canine model. AB - OBJECTIVE: To evaluate the dynamic changes of the power Doppler (PD) in acute renal vein occlusion and recanalization in a canine model. MATERIALS AND METHODS: We performed a PD of the kidney during graded renal vein occlusion and recanalization induced by balloon inflation and deflation in nine dogs. The PD images were transferred to a personal computer, and the PD signals were quantified. RESULTS: We observed the temporal change of the PD signal during renal vein occlusion and recanalization, with a decrease in the PD signal during occlusion and an increase during recanalization. The mean PD signal decreased gradually as the renal vein was occluded, and conversely increased gradually with sequential relief of occlusion. The sequential change of the mean value of the PD signal was statistically significant. CONCLUSION: The PD can detect a change in renal blood flow during acute renal vein occlusion and recanalization in a canine model. The PD may be used as a helpful tool for the early detection of acute renal vein thrombosis and the monitoring of renal perfusion. PMID- 18385560 TI - Correlations between the various methods of estimating prostate volume: transabdominal, transrectal, and three-dimensional US. AB - OBJECTIVE: To evaluate the correlations between prostate volumes estimated by transabdominal, transrectal, and three-dimensional US and the factors affecting the differences. MATERIALS AND METHODS: The prostate volumes of 94 consecutive patients were measured by both transabdominal and transrectal US. Next, the prostate volumes of 58 other patients was measured by both transrectal and three dimensional US. We evaluated the degree of correlation and mean difference in each comparison. We also analyzed possible factors affecting the differences, such as the experiences of examiners in transrectal US, bladder volume, and prostate volume. RESULTS: In the comparison of transabdominal and transrectal US methods, the mean difference was 8.4 +/- 10.5 mL and correlation coefficient (r) was 0.775 (p < 0.01). The experienced examiner for the transrectal US method had the highest correlation (r = 0.967) and the significantly smallest difference (5.4 +/- 3.9 mL) compared to the other examiners (the beginner and the trained; p < 0.05). Prostate volume measured by transrectal US showed a weak correlation with the difference (r = 0.360, p < 0.05). Bladder volume did not show significant correlation with the difference (r = -0.043, p > 0.05). The comparison between the transrectal and three-dimensional US methods revealed a mean difference of 3.7 +/- 3.4 mL and the correlation coefficient was 0.924 for the experienced examiner. Furthermore, no significant difference existed between examiners (p > 0.05). Prostate volume measured by transrectal US showed a positive correlation with the difference for the beginner only (r = 0.405, p < 0.05). CONCLUSION: In the prostate volume estimation by US, experience in transrectal US is important in the correlation with transabdominal US, but not with three-dimensional US. Also, less experienced examiners' assessment of the prostate volume can be affected by prostate volume itself. PMID- 18385561 TI - Radiofrequency ablation using a monopolar wet electrode for the treatment of inoperable non-small cell lung cancer: a preliminary report. AB - OBJECTIVE: To assess the technical feasibility and complications of radiofrequency ablation (RFA) using a monopolar wet electrode for the treatment of inoperable non-small cell lung malignancies. MATERIALS AND METHODS: Sixteen patients with a non-small cell lung malignancy underwent RFA under CT guidance. All the patients were non-surgical candidates, with mean maximum tumor diameters ranging from 3 to 6 cm (mean: 4.6 +/- 1.1 cm). A single 16-gauge open-perfused electrode with a 2 cm exposed tip was used for the procedure. A 0.9% NaCl saline solution was used as the perfusion liquid with the flow adjusted to 30 mL/h. The radiofrequency energy was applied for 10-40 minutes. The response to RFA was evaluated by performing contrast-enhanced CT immediately after RFA, one month after treatment and then every three months thereafter. RESULTS: Technical failure was observed in six (37.5%) of 16 patients: intractable pain (n = 2) and non-stop coughing (n = 4). The mean follow-up interval was 15 +/- 8 months (range: 9-31 months). The mean maximum ablated diameter in the technically successful group of patients ranged from 3.5 to 7.5 cm (mean 5.1 +/- 1.3 cm). Complete necrosis was attained for eight (80%) of 10 lesions, and partial necrosis was achieved for two lesions. There were two major complications (2/10, 20%) encountered: a hemothorax (n = 1) and a bronchopleural fistula (n = 1). CONCLUSION: Although RFA using a monopolar wet electrode can create a large ablation zone, it is associated with a high rate of technical failure when used to treat inoperable non-small cell lung malignancies. PMID- 18385562 TI - Ethanol embolotherapy of pelvic arteriovenous malformations: an initial experience. AB - OBJECTIVE: We retrospectively assessed the results of performing ethanol embolization for pelvis arteriovenous malformations (AVMs). MATERIALS AND METHODS: During the past 10 years, eight patients (8 females, age range: 27-52 years) with AVMs in the pelvic wall (n = 3) and uterus (n = 5) underwent staged ethanol embolizations (range: 1-5, mean: 2.5) under general anesthesia. Ethanol embolization was performed by the use of the transcatheter and/or direct puncture techniques. Clinical follow-up was performed for all of the patients, and imaging follow-up was available for seven patients. The therapeutic outcomes were established by evaluating the clinical outcome of the signs and symptoms, as well as the degree of devascularization observed on post-procedural angiography. RESULTS: During the 20 sessions of ethanol embolization, the solitary transarterial approach was used 14 times, the transvenous approach was used three times and direct puncture was used once. For two patients, the transarterial and transvenous or direct puncture approaches were used together in one session. For four patients, ethanol and coils were used as embolic agents, and n-butyl cyanoacrylate (NBCA) and ethanol were used in one patient. Seven (88%) of eight patients were cured of their AVMs and one patient (12%) displayed improvement. Major complications were seen in two patients (25%). CONCLUSION: Ethanol embolization is effective for the treatment of pelvic arteriovenous malformations, though there is a chance of a major complication. PMID- 18385563 TI - Grading anterior cruciate ligament graft injury after ligament reconstruction surgery: diagnostic efficacy of oblique coronal MR imaging of the knee. AB - OBJECTIVE: The purpose of this study was to evaluate the diagnostic efficacy of using additional oblique coronal MRI of the knee for grading anterior cruciate ligament (ACL) graft injury after ligament reconstruction surgery. MATERIALS AND METHODS: We retrospectively reviewed 51 consecutive MR knee examinations of 48 patients who underwent both ACL reconstruction and follow-up arthroscopy. The MR examinations included the orthogonal axial, sagittal, coronal images and the oblique coronal T2-weighted images, which were oriented in parallel with the course of the femoral intercondylar roof. Two radiologists independently evaluated the status of the ACL grafts with using the routine knee MRI and then with adding the oblique coronal imaging. The severity of ACL graft injury was graded using a 3-point system from MR images as intact, partial tear or complete tear, and the results were compared with the arthroscopic results. Weighted kappa statistics were used to analyze the diagnostic accuracies of the knee MRI with and without the additional oblique coronal imaging. For each evaluation, the observers reported a confidence level for grading the ACL graft injuries in the two imaging groups. RESULTS: The weighted kappa values according to the routine knee MRI were 0.555 (reader 1) and 0.515 (reader 2). The inclusion of additional oblique coronal imaging increased the weighted kappa values to 0.666 (reader 1) and 0.611 (reader 2). The mean confidence levels by each reader were significantly higher (p < 0.01, paired t-test) with the additional oblique coronal imaging than by using the routine knee MRI alone. CONCLUSION: The additional use of oblique coronal MRI of the knee improves both the diagnostic accuracy and confidence for grading ACL graft injury. PMID- 18385564 TI - Intrapancreatic accessory spleen: findings on MR Imaging, CT, US and scintigraphy, and the pathologic analysis. AB - Although the tail of the pancreas is the second most common site of an accessory spleen, intrapancreatic accessory spleen (IPAS) has rarely been noted radiologically. However, as the imaging techniques have recently advanced, IPAS will be more frequently detected as an incidental pancreatic nodule on CT or MRI. Because accessory spleens usually pose no clinical problems, it is important to characterize accessory spleens as noninvasively as possible. An IPAS has similar characteristics to those of the spleen on the precontrast and contrast-enhanced images of all the imaging modalities. In particular, inhomogeneous enhancement of an IPAS in its early phases may be a diagnostic clue. Superparamagnetic iron oxide (SPIO)-enhanced MRI and Levovist-enhanced US, and the mechanisms of which are theoretically similar to that of Tc-99m scintigraphy, can be used as alternative tools to confirm the diagnosis of IPAS. An IPAS shows a significant signal drop similar to the spleen on the SPIO-enhanced T2 or T2*-weighted imaging and prolonged enhancement on the delayed hepatosplenic phase of contrast-enhanced US. We review and illustrate the differential points between IPAS and hypervascular pancreatic tumors in this manuscript. PMID- 18385565 TI - Reversible lansoprazole-induced interstitial lung disease showing improvement after drug cessation. AB - Lansoprazole is an acid proton-pump inhibiting drug that is used for the treatment of duodenal or gastric ulcers, H. pylori infection, gastroesophageal reflux disease or Zollinger-Ellison syndrome. Although lansoprazole is well known for its gastrointestinal and dermatologic adverse effects, mild pulmonary symptoms are also known to develop from taking this drug. There have been no reports about lansoprazole-induced interstitial lung disease. We report here a case of lansoprazole-induced interstitial lung disease that developed in a 66 year-old man. PMID- 18385566 TI - Pulsed and color Doppler sonographic findings of penile Mondor's disease. AB - This report describes the color and pulsed Doppler US findings of penile Mondor's disease. The pulsed Doppler US findings of penile Mondor's disease have not been previously published, so we report here for the first time on the cavernosal arterial flow signal pattern of penile Mondor's disease. Penile Mondor's disease is rare disease that's characterized by thrombosis in the dorsal vein of the penis. The previous reports on penile Mondor's disease are concerned with the color Doppler US finding without the flow signals in this area, but these findings are insufficient to understand the hemodynamics in penile Mondor's disease. We report for the first time on a cavernosal artery flow signal pattern of low peak systolic velocity and high-resistance. PMID- 18385567 TI - Embolization of a bleeding maxillary arteriovenous malformation via the superficial temporal artery after external carotid artery ligation. AB - We report a new approach of embolization in a 15-year-old boy that presented with a massive hemorrhage from a maxillary arteriovenous malformation. Re-bleeding occurred after emergent ligation of the external carotid artery. The bleeding was successfully controlled by embolization via the superficial temporal artery. PMID- 18385570 TI - Assessing the total patient. PMID- 18385568 TI - Atypical radiological manifestation of pulmonary metastatic calcification. AB - Metastatic pulmonary calcification refers to calcium deposition in the normal pulmonary parenchyma and this deposition is secondary to abnormal calcium metabolism. The most common radiologic manifestation consists of poorly-defined nodular opacities that are mainly seen in the upper lung zone. We present here a case of metastatic pulmonary calcification that manifested as atypical, dense, calcium deposition in airspaces within the previously existing consolidation in the bilateral lower lobes, and this process was accelerated by pneumonia complicated sepsis in a patient with hypercalcemia that was due to hyperparathyroidism. PMID- 18385571 TI - Wound care expert: a term of uncertainty. PMID- 18385575 TI - 2008 coding and payment changes for physicians. PMID- 18385576 TI - The status of wound care in Brazil. PMID- 18385577 TI - A prospective, randomized, multisite clinical evaluation of a transparent absorbent acrylic dressing and a hydrocolloid dressing in the management of Stage II and shallow Stage III pressure ulcers. AB - OBJECTIVE: To compare clinical performance of a transparent absorbent acrylic dressing (3M Tegaderm Absorbent Clear Acrylic Dressing ]TAAD[; 3M Company, St Paul, MN) and a hydrocolloid dressing (HD ]DuoDERM CGF, ConvaTec, ER Squibb & Sons, Princeton, NJ[) in the management of Stage II and shallow Stage III pressure ulcers. DESIGN: Prospective, open-label, randomized, comparative, multisite clinical evaluation. Patients were followed up for a maximum of 56 days or until their ulcer healed. At weekly intervals, investigators conducted wound assessments and dressing performance evaluations. SETTING: Wound care clinics, home care, and long-term care. PATIENTS: Thirty-five patients received the TAAD, and 37 received the HD. OUTCOME MEASURES: Dressing performance assessments, patient comfort, dressing wear time, and wound healing were measured. RESULTS: The majority of investigator assessments favored the TAAD. Considerations given included the ability to center dressings over the ulcer (P = .005), ability to assess the ulcer before (P < .001) and after (P < .001) absorption, barrier properties (P = .039), patient comfort during removal (P < .001), overall patient comfort (P = .048), conformability before (P = .026) and after (P = .001) absorption, ease of removal (P < .001), nonadherence to wound bed (P < .001), residue in the wound (P = .002), residue on periwound skin (P < .001), and odor after absorption (P = .016). Overall satisfaction favored the TAAD (P < .001), and a high value was placed on its transparent feature (P < .001). Mean (SD) wear time for the TAAD was 5.7 (2.55) days compared with 4.7 (2.29) days for the HD (P = .086). This 1-day difference in wear time was clinically noticeable by the investigators (P = .035). Wound closure for the 2 dressing groups was nearly identical (P = .9627). CONCLUSIONS: Performance results favored the TAAD over the HD as standard treatment for Stage II and shallow Stage III pressure ulcers. PMID- 18385578 TI - Chronic wound pain: a conceptual model. AB - PURPOSE: To present the wound care practitioner with a model for the assessment and treatment of wound-related pain. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in wound care and related disorders. OBJECTIVES: After reading this article and taking this test, the reader should be able to: 1. Discuss the pathophysiology of chronic pain and the wound pain model. 2. Describe the patient's wound-related pain perspective. 3. Identify aspects of local wound care and their relationship to pain. PMID- 18385580 TI - Autonomous critical care for the austere environment. Introduction. PMID- 18385581 TI - Closed-loop strategies for patient care systems. AB - Military operations, mass casualty events, and remote work sites present unique challenges to providers of immediate medical care, who may lack the necessary skills for optimal clinical management. Moreover, the number of patients in these scenarios may overwhelm available health care resources. Recent applications of closed-loop control (CLC) techniques to critical care medicine may offer possible solutions for such environments. Here, feedback of a monitored variable or group of variables is used to control the state or output of a dynamic system. Some potential advantages of CLC in patient management include limiting task saturation when there is simultaneous demand for cognitive and active clinical intervention, improving quality of care through optimization of the titration of medications, conserving limited consumable supplies, preventing secondary insults in traumatic brain injury, shortening the duration of mechanical ventilation, and achieving appropriate goal-directed resuscitation. The uses of CLC systems in critical care medicine have been increasingly explored across a wide range of therapeutic modalities. This review will provide an overview of control system theory as applied to critical care medicine that must be considered in the design of autonomous CLC systems, and introduce a number of clinical applications under development in the context of deployment of such applications to austere environments. PMID- 18385583 TI - Autonomous control of ventilation. PMID- 18385582 TI - Autonomous control of oxygenation. PMID- 18385584 TI - Closed-loop and decision-assist resuscitation of burn patients. AB - Effective resuscitation is critical in reducing mortality and morbidity rates of patients with acute burns. To this end, guidelines and formulas have been developed to define infusion rates and volume requirements during the first 48 hours postburn. Even with these standardized resuscitation guidelines, however, over- and under-resuscitation are not uncommon. Two approaches to adjust infusion rate are decision-assist and closed-loop algorithms based on levels of urinary output. Specific decision assist guidelines or a closed-loop system using computer-controlled feedback technology that supplies automatic control of infusion rates can potentially achieve better control of urinary output. In a properly designed system, closed-loop control has the potential to provide more accurate titration rates, while lowering the incidence of over- and under resuscitation. Because the system can self-adjust based on monitoring inputs, the technology can be pushed to environments such as combat zones where burn resuscitation expertise is limited. A closed-loop system can also assist in the management of mass casualties, another scenario in which medical expertise is often in short supply. This article reviews the record of fluid balance of contemporary burn resuscitation and approaches, as well as the engineering efforts, animal studies, and algorithm development of our most recent autonomous systems for burn resuscitation. PMID- 18385585 TI - Closed-loop control of fluid therapy for treatment of hypovolemia. AB - Closed-loop algorithms and resuscitation systems are being developed to control IV infusion rate during early resuscitation of hypovolemia. Although several different physiologic variables have been suggested as an endpoint to guide fluid therapy, blood pressure remains the most used variable for the initial assessment of hemorrhagic shock and the treatment response to volume loading. Closed-loop algorithms use a controller function to alter infusion rate inversely to blood pressure. Studies in hemorrhaged conscious sheep suggest that: (1) a small reduction in target blood pressure can result in a significant reduction in volume requirement; (2) nonlinear algorithms may reduce the risk of increased internal bleeding during resuscitation; (3) algorithm control functions based on proportional-integral, fuzzy logic, or nonlinear decision tables were found to restore and maintain blood pressure equally well. Proportional-integral and fuzzy logic algorithms reduced mean fluid volume requirements compared with the nonlinear decision table; and (4) several algorithms have been constructed to the specific mechanism of injury and the volume expansion properties of different fluids. Closed-loop systems are undergoing translation from animal to patient studies. Future smart resuscitation systems will benefit from new noninvasive technologies for monitoring blood pressure and the development of computer controlled high flow intravenous pumps. PMID- 18385586 TI - Physiological and medical monitoring for en route care of combat casualties. AB - BACKGROUND: Most prehospital medical interventions during civilian and military trauma casualty transport fail to utilize advanced decision-support systems for treatment and delivery of medical interventions, particularly intravenous fluids and oxygen. Current treatment protocols are usually based on standard vital signs (eg, blood pressure, arterial oxygen saturation) which have proven to be of limited value in detecting the need to implement an intervention before cardiovascular collapse. A primary objective of the US Army combat casualty care research program is to reduce mortality and morbidity during casualty transport from the battlefield through advanced development of a semiautomated decision support capability for closed-loop resuscitation and oxygen delivery. METHODS: To accomplish this goal, the Trauma Informatics Research Team at the US Army Institute of Surgical Research has developed two models for evidence-based decision support 1) a trauma patient database for capture and analysis of prehospital vital signs for identification of early, novel physiologic measurements that could improve the control of closed-loop systems in trauma patients; and, 2) a human experimental model of central hypovolemia using lower body negative pressure to improve the understanding and identification of physiologic signals for advancing closed-loop capabilities with simulated hemodynamic responses to hemorrhage. RESULTS: In the trauma patient database and lower body negative pressure studies, traditional vital sign measurements such as systolic blood pressure and oxygen saturation fail to predict mortality or indicate the need for life saving interventions or reductions in central blood volume until after the onset of cardiovascular collapse. We have evidence from preliminary analyses, however, that indicators of reduced central blood volume in the presence of stable vital signs include 1) reductions in pulse pressure; 2) changes in indices of autonomic balance derived from calculation of heart period variability (ie, linear and non-linear analyses of R-R intervals); and 3) reductions in tissue oxygenation. CONCLUSIONS: We propose that derived indices based on currently available technology for continuous monitoring of specific hemodynamic, autonomic, and/or metabolic responses could provide earlier recognition of hemorrhage than current standard vital signs and allow intervention before the onset of circulatory shock. Because of this, such indices could provide improved feedback for closed-loop control of patient resuscitation and oxygen delivery. These technological advances could prove instrumental in advancing decision-support capabilities for prehospital trauma care during transport to higher levels of care in both the military and civilian environments. PMID- 18385587 TI - Autonomous medical care for exploration class space missions. AB - The US-based health care system of the International Space Station contains several subsystems, the Health Maintenance System, Environmental Health System and the Countermeasure System. These systems are designed to provide primary, secondary and tertiary medical prevention strategies. The medical system deployed in low Earth orbit for the International Space Station is designed to support a "stabilize and transport" concept of operations. In this paradigm, an ill or injured crewmember would be rapidly evacuated to a definitive medical care facility (DMCF) on Earth, rather than being treated for a protracted period on orbit. The medical requirements of the short (7 day) and long duration (up to 6 months) exploration class missions to the moon are similar to low Earth orbit class missions but also include an additional 4 to 5 days needed to transport an ill or injured crewmember to a DMCF on Earth. Mars exploration class missions are quite different in that they will significantly delay or prevent the return of an ill or injured crewmember to a DMCF. In addition the limited mass, power and volume afforded to medical care will prevent the mission designers from manifesting the entire capability of terrestrial care. National Aeronautics and Space Administration has identified five levels of care as part of its approach to medical support of future missions including the Constellation program. To implement an effective medical risk mitigation strategy for exploration class missions, modifications to the current suite of space medical systems may be needed, including new crew medical officer training methods, treatment guidelines, diagnostic and therapeutic resources, and improved medical informatics. PMID- 18385588 TI - NAON's gateway is open for orthopaedic excellence! PMID- 18385589 TI - Nurse-physician collaboration. PMID- 18385590 TI - Developing and implementing an orthopaedic outcomes program: a step-by-step guide. AB - Evidence-based nursing practice provides the highest quality of patient care. In an orthopaedic setting, the study of outcomes across a patient population offers the ability to determine associations between the treatment regime and function, complication rates, patient satisfaction, and quality of life as evidence for practice. These associations give the orthopaedic nursing community valuable insight into patient results. An outcomes database can both assist nurses in assessing the results of their nursing interventions and in providing the basis for submitting papers to journals and scientific symposiums. These papers can provide information and education for establishment of an evidence-based nursing practice. This article describes the planning, development, and implementation of a comprehensive orthopaedic outcomes program including nursing intervention outcomes at one institution. We offer a step-by-step guide based on our experience for other institutions that plan to initiate an outcomes program with inclusion of nursing outcomes. PMID- 18385592 TI - Avoiding the impact of musculoskeletal pain on quality of life in children with hemophilia. AB - The most frequent, severe clinical manifestation of hemophilia is hemarthrosis, which often develops at a young age and is initiated by repeated joint bleeds. Blood within a joint results in inflammation and hypertrophy of synovial membranes, causing increased vascularization of the joint and bone degeneration. In addition, hemarthrosis is associated with pain, impaired mobility, and reduced health-related quality of life (HRQoL). Although data on the effect of joint pain on pediatric HRQoL are currently lacking, both health-related outcome and treatment-experience studies reported the benefits of early identification, assessment, and treatment of pain in improving patient outcome and HRQoL. Ultimately, prevention of bleeding through aggressive treatment of joint bleeds and prophylaxis with factor concentrates are key to ensuring the best musculoskeletal outcome for pediatric patients with hemophilia. PMID- 18385594 TI - A study of naloxone effect on urinary retention in the patient receiving morphine patient-controlled analgesia. AB - PURPOSE: The purpose of the study was to compare urinary retention rates following orthopaedic surgery in patients who received low-dose intravenous naloxone while receiving morphine patient-controlled analgesia with patients who did not receive naloxone. DESIGN: Randomized controlled trial without blinding. SAMPLE: There were 97 participants consenting to the study, 45 were randomly assigned to a control group and 52 assigned to an experimental group. Forty-three patients in the control group and 47 in the experimental group (90 total) completed the study protocol. FINDINGS: Postoperative urinary residuals were lower, patients voided more frequently, and fewer catheterizations were needed when given low-dose naloxone while receiving morphine patient-controlled analgesia. At the same time, naloxone in small doses was found to have negligible effect on overall patient pain control. PMID- 18385595 TI - Randomized controlled trials: methodological concepts and critique. AB - Randomized controlled trials, also known as true experiments or intervention studies, are considered to be the gold standard research design for demonstrating a cause-and-effect relationship between an intervention and an outcome. This article will describe key methodological concepts that make the randomized controlled trial this gold standard. Practicing from an evidence-based perspective requires practitioners to be able to critique the strengths and weaknesses of a research study in order to make decisions about adoption of the intervention into one's practice area. Key components to the critique of a randomized controlled trial are defined and the process is illustrated by a critique of Gallo and colleagues article, A Study of Naloxone Effect on Urinary Retention in the Patient Receiving Morphine PCA, published in this issue. PMID- 18385597 TI - Transforming your professional self: encouraging lifelong personal and professional growth. AB - Transforming from student nurse to registered nurse is often discussed in a capstone class or a hospital orientation program. Changes in professional plans later in the career continuum often occur, but are not always planned. This article discusses the challenges of change, the need for career planning, stages of role acquisition, role socialization, and role transformation. In addition, it outlines the importance of creating a career plan to meet future career goals. PMID- 18385599 TI - Conflict of interest: nurses at risk! AB - Conflict of interest as it relates to healthcare is gaining increasing attention. Pharmaceutical companies and manufacturers that produce medical devices are coming under greater scrutiny because of the influence that their marketing practices may have on the patient management decisions made by healthcare professionals. The result is that healthcare agency administrators are developing conflict of interest policies and procedures for their professional employees. The driving force behind many of these policies is the need to maintain the trust of the public by refraining from questionable professional conduct. This article presents 2 hypothetical cases to provide nurses with an understanding of the concept of conflict of interest and the ethical considerations this issue raises, and describes the subtle and not-so-subtle influences on professional practice decisions. Recommendations are offered to help nurses avoid conflict of interest and preserve their professional integrity. It is incumbent upon nurses to become cognizant of the types of situations that may present a conflict of interest for them and to take the necessary steps to avoid such professional impropriety. PMID- 18385600 TI - Colles' fracture. AB - Many people "slip and fall", especially in the icy areas of the winter season. To prevent an injury to the head, most people put their hand out to hit the ground first, so the wrist usually gets injured. The most frequent injury from this type of "intervention" is a fracture to the distal radius and/or ulna, which is frequently called a "Colles' fracture." PMID- 18385603 TI - Rifaximin in treatment of recurrent Clostridium difficile-associated diarrhea: an uncontrolled pilot study. PMID- 18385605 TI - Liver disease from asymptomatic constrictive pericarditis. AB - Congestive hepatopathy is a known complication of cardiac disease and is typically identified in the context of an established cardiac diagnosis and profound cardiopulmonary symptoms. We report the case of a 28-year-old man with liver disease secondary to asymptomatic constrictive pericarditis. This case highlights the need for gastroenterologists to consider occult cardiac disease as a cause of unexplained liver dysfunction. PMID- 18385604 TI - High risk of malignancy in familial Barrett's esophagus: presentation of one family. AB - Barrett's esophagus is an acquired condition fundamentally related to the presence of severe and prolonged pathologic acid and biliary gastro-esophageal reflux. However, genetic factors may also play a role in some cases. The aim of this study is to present 3 generations of a Spanish family with the largest number of members so far reported with Barrett's esophagus or esophageal adenocarcinoma. Of the 24 members of this family studied over 3 generations, 6 patients developed esophageal adenocarcinoma, 4 Barrett's esophagus, 6 clinical symptoms of gastro-esophageal reflux disease without Barrett's esophagus, and 8 were asymptomatic. In conclusion, patients with familial Barrett's esophagus get the disease more severely with a high rate of malignancy and, therefore, the endoscopic surveillance should be closer than in cases of nonfamilial Barrett's esophagus. PMID- 18385606 TI - Primary liver carcinoma exhibiting dual hepatocellular-biliary epithelial differentiations associated with citrin deficiency: a case report. AB - We report a 50-year-old male patient with primary liver carcinoma exhibiting dual hepatocellular and biliary epithelial differentiations associated with citrin deficiency (asymptomatic adult-onset type II citrullinemia, CTLN2). Although so far 14 CTLN2 patients with hepatocellular carcinoma have been reported, this report describes a unique case of liver carcinoma showing the features of both hepatocellular and cholangiocellular carcinoma. In addition to the clinical data of the 14 patients reported previously, the findings in our patient suggest that the citrin deficiency might be one of the key disorders causing hepatocellular carcinoma especially at younger ages and can also play an important role in hepatocarcinogenesis of the hepatic progenitor cells, which have the bipotential to differentiate into both hepatocytes and cholangiocytes. PMID- 18385622 TI - Outcome predictors of rehabilitation for first stroke in the elderly. AB - AIM: The prediction of stroke outcome in the elderly can be rather difficult, due to the potential interference into disability and handicap development of several clinical modifiers, such as comorbidity, medical complications, neuropsychological impairment proper to the aging brain and social issues. These factors can strongly affect old patient response to rehabilitation and need to be taken into account, along with ageing per se, to optimize health resource efficiency for the care of disability due to stroke. In this study, we tried to identify outcome determinants of stroke rehabilitation specific for the elderly. METHODS: A total of 359 first-stroke patients aged 75 years, admitted for active rehabilitation treatment to hospital rehabilitation wards, were enrolled into a multicenter cohort (prospective) study. They all underwent a comprehensive medical rehabilitation program. We considered as primary outcomes the frequency of home discharge and the extent of functional recovery, assessed by Functional Independence Scale (FIM) and expressed as the Montebello Rehabilitation Factor Score (MRFS) efficacy. Each measure of outcome was related to age, as well as other potential clinical and functional confounders, according to a multivariate model. For each dependent variable, two models were developed, using either FIM total score or FIM domains scores at admission among predictors. RESULTS: FIM total score increased from 55.8+/-24 to 75.3+/-30 (P<0.0001), with a mean MRFS efficacy of 0.33+/-0.25. Most patients (79.9%) were discharged home. Age turned out as independently and inversely related to MRFS, explaining at the most 3.6% of its variance, although FIM at admission was its most powerful predictor. Home discharge was not related to age, but to social issues, such as living in family before admission, and cognition. CONCLUSION: The present study suggests that rehabilitation can be effective in elderly stroke patients, in improving function as well as in favorably affecting discharge destination. In fact, age per se predicts the outcome at a lesser extent than other clinical covariates, such as functional and cognitive status at admission and social situation. PMID- 18385623 TI - Depression and functional outcome after stroke: the effect of antidepressant therapy on functional recovery. AB - AIM: The optimal strategy to prevent post-stroke depression is an important but still-unresolved issue. This study examined the differences in functional recovery among post-stroke depressed patients (DP) compared to post-stroke non depressed patients (NDP) over the course of six months after stroke. METHODS: On the basis of a semistructured psychiatric examination, DSM-IV diagnostic criteria and the Hamilton Depression score, a consecutive series of ischemic or hemorrhagic stroke patients were included in this study. They had suffered from first-time stroke, and did not have depression diagnosis before. During follow up, treatment with 20 mg/day citalopram per os was initiated whenever a diagnosis of depression was established. Their functional recoveries were assessed using the Scandinavian Stroke Scale, the modified Rankin scale and the Barthel index during acute hospitalization, at the time of depression diagnosis and at the third and sixth month follow-up visits. RESULTS: Forty patients met the diagnostic criteria and 11 patients suffered from depression during the follow-up period. There were no differences in demographic variables, lesion characteristics and neurological symptoms between DP and NDP. Functional recovery in DP was impaired in comparison to NDP (P<0.05). All DP, whose mood improved after administration of citalopram, and improved daily functions living functions during the follow-up. CONCLUSION: This study's findings suggest that remission of post-stroke depression is associated with improvement in functional recovery. Early diagnosis and effective treatment of depression will help the rehabilitation outcome of stroke patients. PMID- 18385624 TI - The effect of balance training on motor recovery and ambulation after stroke: a randomized controlled trial. AB - AIM: To investigate the effects of balance training, using force platform biofeedback, on motor recovery, mobility and activity level of hemiparetic patients with stroke. METHODS: In this randomized, controlled, assessor-blinded trial 41 inpatients [mean (+/-SD) age of 60.9 (+/-11.7) years] with hemiparesis after stroke (median time since stroke 6 months) were randomly assigned to an experimental or a control group. The control group (n=19) participated in a conventional stroke inpatient rehabilitation program, whereas the experimental group (n=22) received 15 sessions of balance training (using force platform biofeedback) in addition to the conventional program. Main outcome measures were motor recovery of the lower extremity (Brunnstrom staging), mobility (Rivermead Mobility Index, RMI) and activity level (Functional Independence Measure, FIM) that performed one week before and after the experimental treatment program. RESULTS: Both groups were similar in terms of baseline clinical characteristics. Motor recovery, mobility and activity level improved significantly in both groups (P<0.05). Between-group difference of mean change score was not significant for the Brunnstrom stages (0.23 vs 0.26), RMI (2.9 vs 2.2) and FIM score (10.7 vs 11.5). CONCLUSION: In our group of stroke patients, balance training combined with a conventional rehabilitation program does not provide additional benefit in terms of lower extremity motor recovery, mobility and activity level. PMID- 18385625 TI - Poor compliance with ankle-foot-orthoses in Charcot-Marie-Tooth disease. AB - AIM: The aim of this study is to evaluate the compliance with ankle-foot orthoses (AFOs) in patients previously prescribed and affected with Charcot-Marie-Tooth disease (CMT). METHODS: Twenty-five Italian patients (8 males 17 females; mean age: 41.6 years, range 16-54) with severe bilateral footdrop (leg-sole angle alpha >105 degrees ) alone or associated with other problems (rotation, plantarflexor failure, knee flexor failure) were examined by a physiatrist (with measurement of the leg-sole angle alpha' with their footwear) and interviewed by a psychologist. RESULTS: Only 5 patients (20%) used AFOs (3 prefabricated polypropylene AFOs, 2 custom-made short AFOs incorporated in high-top boots) with satisfactory functional results (alpha' <=94 degrees ; reported increased mobility and no more falls). The interview revealed that all patients had a bad relationship with their own body. The 3 subjects using prefabricated AFOs said that they hated them and one of them complained of pain. Patients not using AFOs justified their decision with statements such as: ''I am not yet ready to accept them'' (n=3) or ''I can still manage without them for a while'' (n=2) or both (n=15). Four patients had experienced pain during the trial, 2 had not found proper shoes to accommodate them and 12 were absolutely not interested in AFOs and, therefore, had not gone to an orthotist. CONCLUSION: Compliance with AFOs is poor. Patients with CMT discard AFOs because they highlight their disability, are not essential for their limited daily walking and are uncomfortable. We suggest that prescription of AFOs be accompanied with psychological support and that research of more comfortable and cosmetically acceptable solutions for the problem of footdrop be stimulated. PMID- 18385626 TI - Weight excess of school materials and its risks factors in South Brazil. A cross sectional study. AB - AIM: There is little information about weight excess of school materials in Brazil. The aim of this study was to determine the prevalence and risk factors of excess weight of school materials in a private and a public school in the city of Pelotas, Brazil. METHODS: A cross-sectional study was performed to examine students' equipment in both a private and a public school. Demographic data were collected and children were weighed and measured. The types of schoolbag, notebook, snack, and other materials taken to school were identified and weighed separately from Monday to Friday. An excess weight of school material was defined for schoolbags weighing more than 10% of each student's weight. RESULTS: The study included 226 (48.8%) students from a private school and 237 (51.2%) students from a public school. From this sample, 38.2% of the total students carried an excess weight of school materials, 68.5% (155 children) of whom were private school students compared to 9.3% (22 children) of public school students, giving a prevalence ratio (PR) of 7.4 (CI95%, 4.9-11.1). After differentiating the sample in terms of school type, the association between weight excess of school material and school bag type (trolley pack), notebook type (hard back or spiral notebook) and transport of snacks were positively reported. CONCLUSION: Due to the higher income of the families of private school children, the type of school equipment carried was more costly and was the heaviest. These data should be considered in educational campaigns in order to reduce school equipment weight. PMID- 18385627 TI - Caregiver strain in Parkinson's disease and the impact of disease duration. AB - AIM: The task of managing care for patients with Parkinson's disease (PD) often falls upon a family member taking on the role as a caregiver (CG) implying a burden on these CGs. The aim of this study was to evaluate CG strain of PD patients with regarding different psychosocial domains and the influence of PD/CG duration of PD. METHODS: A cross-sectional telephone interview survey of 451 CGs randomly selected from the registry of the Swedish Parkinson's Disease Association. A structured questionnaire covering sociodemographic, psychosocial, and general CG factors, sleep and depression of the CG as well as issues of the patient's disease was used by 4 independent interviewers blinded to the study objective. RESULTS: Four hundred and four of 451 (90%) CGs responded with a mean age of 68.5 years with 62% females. The results were stratified in 3 groups with regard to disease duration of the PD patient, 0-4, 5-10, and >11 years, respectively. General health condition of the CGs was regarded satisfactory independent of disease duration. Insufficient sleep and disease related stress were considered to be prominent in 36% and 61%, respectively, being significantly more prominent in the group with the longest disease duration. Decreased mood was reported in 31% with no difference between groups. More than 30% of CGs also experienced daily problems with tiredness and sleep disturbance; 27% hypertension; 17% muscle strain, headache and fatigue; and 14% gastro-intestinal problems most items regardless of disease duration. The most troublesome symptoms of the patients to the CGs were reported to be the motor dysfunction (58%). More than half experienced little or no understanding of their situation. CONCLUSION: CGs are afflicted with strain and burden in many psychosocial and somatic domains despite satisfactory general wellbeing independent of disease duration. The longer disease duration, and, accordingly CG duration, the more impact on certain domains of CG burden, however, with little understanding of their situation. These findings should be given greater consideration when organizing and planning for PD care in the health care system and the community. PMID- 18385628 TI - Pelvic floor muscle training versus no treatment for urinary incontinence in women. A Cochrane systematic review. AB - BACKGROUND: Pelvic floor muscle training is the most commonly used physical therapy treatment for stress urinary incontinence. It is sometimes recommended for mixed and less commonly for urge urinary incontinence. OBJECTIVES: The aim of this paper was to determine the effects of pelvic floor muscle training for women with urinary incontinence in comparison to no treatment, placebo or sham treatments, or other inactive control treatments. METHOD: The Cochrane Incontinence Group Specialized Trials Register was searched up until December 1, 2004. The review included randomized or quasi-randomized trials in women with stress, urge or mixed urinary incontinence. One arm of the trial comprised pelvic floor muscle training, the other comprised either no treatment, placebo, sham, or other inactive control treatment. The trials were independently assessed for eligibility and methodological quality. Data were extracted then cross-checked by the two authors. Disagreements were resolved by discussion. The data were processed as described in the Cochrane Handbook. The trials were sub-grouped by diagnosis. Formal meta-analysis was not undertaken because of study heterogeneity. RESULTS: Thirteen trials involving 714 women met the inclusion criteria; however, only six trials (403 women) contributed to data analysis. CONCLUSION: Overall, the review provides support for the widespread recommendation that pelvic floor muscle training be included in first-line conservative management programs for women with stress, urge or mixed urinary incontinence. PMID- 18385629 TI - Cochrane reviews: evidence in rehabilitation. PMID- 18385630 TI - Determinants of disability in chronic musculoskeletal health conditions: a literature review. AB - Knowledge of the determinants of disability in musculoskeletal conditions (MSC) is critical for reducing their burden. No epidemiologic studies from a truly comprehensive perspective consider environmental factors (EF) and personal factors (PF) as determinants of disability. However, one can identify candidate EF from the International Classification of Functioning, Disability and Health (ICF) Core Sets for rheumatoid arthritis (RA), osteoporosis (OP), osteoarthritis (OA), low back pain (LBP) and chronic wide spread pain (CWP). The objective of this literature review was to contribute to the validation of the EF from the ICF Core Sets for MSC and the candidate PF from a (ICF) Delphi exercise, as well as from the report of the Bone and Joint Decade (BJD) Health Strategy Project. The results of the literature search focus on reviews published between January 1991 and March 2006 that contained information on EF and PF that determine disability in LBP, RA and OA. Many PF and EF included in the ICF Core Sets were confirmed as potential determinants of disability. However, regarding some contextual factors, in particular EF referring to the physical environment, there is a lack of reviews and clinical studies that have investigated their relevance to disability. The predominant medical model in studies on disability in MSC may explain this lack of evidence. However, the increasing attention given to the integrative model of functioning, disability and health of the World Health Organization (WHO) and the approval of the ICF by the World Health Assembly in 2001 may stimulate future research on the effect of EF and PF on disability. PMID- 18385631 TI - Heterotopic ossification due to neuromuscular blocking agent in adult respiratory distress syndrome. A case report. AB - Heterotopic ossification (HO) is the formation of mature bone in soft tissues, which can be seen either in cases of traumatic injury or nontraumatic conditions. HO is more commonly observed after a traumatic event, such as a spinal cord injury or head injury. Although less common, it can also be seen after nontraumatic neurological conditions, such as long term coma or neuromuscular blockade. Patients with adult respiratory distress syndrome (ARDS) are at risk of developing HO, due to a prolonged mechanical ventilation period and exposure to neuromuscular blocking agents. In this case report, the authors describe a 51 year-old woman with ARDS, whose HO developed after long-term sedation with neuromuscular blockade agents. PMID- 18385632 TI - Intrathecal baclofen bolus complicated by deep vein thrombosis and pulmonary embolism. A case report. AB - The aim of this study was to present a case of deep vein thrombosis (DVT) and pulmonary embolism (PE) after an intrathecal baclofen bolus test. The study was carried out at a tertiary care rehabilitation center in Italy, and it reports a case of a 65-year-old woman affected by primary lateral sclerosis, with lower limb spasticity, who underwent an intrathecal baclofen bolus trial. After a 100 microg baclofen test, she developed hypotension and oxygen desaturation lasting two days. A Doppler ultrasonography was positive for DVT, and echocardiography confirmed the diagnosis of PE. A Doppler ultrasonography performed before the test was negative. This is the first report in the literature of DVT and PE after an intrathecal baclofen bolus trial. Clinicians and patients should be informed that the intrathecal baclofen bolus trial procedure could be associated with rare but important risks such as DVT and PE, and they should be alert in recognizing potentially life-threatening complications. PMID- 18385633 TI - Congenital lymphedema of the upper extremity. A case report. AB - This study presents an unusual case of primary lymphedema of the upper extremity in a healthy 28-year-old woman. The onset of swelling of the left upper extremity was observed at birth, but was not accurately diagnosed until the patient visited our department. Diagnostic assessment included evaluating the patient's history and laboratory and radiological data, which were all normal except for the swollen upper extremity and the lymphoscintigraphy findings. The patient was diagnosed as suffering from primary lymphedema of her left arm. Complete decongestive therapy was done and her swelling mildly improved. PMID- 18385634 TI - Orthopedic shoes improve gait in Friedreich's ataxia: a clinical and quantified case study. AB - Our aim was to evaluate with modern tools the efficacy of orthopedic shoes on gait disorders in Friedreich's ataxia. The case of a 26-year-old woman with Friedreich's ataxia is described. She mainly complained of fatigability, ankle instability, frequent falls and pain. Impairments involved a cerebellar syndrome, a proprioceptive deficit, an upper motor neurone syndrome and osteoarticular deformities. Gait disabilities included ataxia and requirement of a cane. Handicap concerned outings, altering quality of life. Orthopedic shoes combined with physical therapy were prescribed. Assessment of treatment was planned after one month. Self-assessment by the patient was noted. Clinical assessment was provided by physical examination and clinical gait analysis supported by video. Quantified assessment was performed with a Gaitrite system recording spatiotemporal gait parameters. Our results demonstrated that orthopedic shoes improved gait disorders in this patient with Friedreich's ataxia. Pain decreased, walking distance increased, falls were less frequent, going out became possible, stability was better, speed, step length and cadence increased. Both clinical and quantified assessment confirmed functional improvement felt by the patient. In conclusion complete medical and social assessment determines quality of prescription in physical and rehabilitation medicine. PMID- 18385635 TI - The role of animal-assisted therapy in physical and rehabilitation medicine. PMID- 18385636 TI - Basic assessment of stroke. Protocollo di minima per l'ictus cerebrale (PMIC). PMID- 18385637 TI - Ariadne's thread: biomedical gateways to not get lost in the Internet labyrinth. AB - The more the Internet grows, the more a researcher has to cope with the so called ''information overload'', or ''data deluge''. Reliability, updating, authoritativeness of a Web site ought to be a requirement, all the more reason for the biomedical field. During the past years libraries and information specialists have been developing projects, such as gateways and virtual reference desks, aimed at supplying trustworthy health information in Web sites. Purpose of this contribution is to present these tools, in order to improve one's search skills. PMID- 18385638 TI - European Journal of Physical and Rehabilitation Medicine: a new clinical rehabilitation journal, an update of our 44 year old Latin name (Europa Medicophysica). PMID- 18385639 TI - Transplantation-transmitted tuberculosis--Oklahoma and Texas, 2007. AB - Approximately 28,000 organ transplants were performed in the United States in 2007. When infections are transmitted from donors, the implications can be serious for multiple recipients. Tuberculosis (TB), a known infectious disease complication associated with organ transplantation, occurs in an estimated 0.35% 6.5% of organ recipients in the United States and Europe posttransplantation. In 2007, the Oklahoma State Department of Health identified Mycobacterium tuberculosis in an organ donor 3 weeks after the donor's death. This report summarizes results of the subsequent investigation, which determined that disseminated TB occurred in two of three transplant recipients from this donor, and one recipient died. Genotypes of the donor and recipient TB isolates were identical, consistent with transmission of TB by organ transplantation. To reduce the risk for TB transmission associated with organ transplantation, organ recovery personnel should consider risk factors for TB when assessing all potential donors. In addition, clinicians should recognize that transplant recipients with TB might have unusual signs or symptoms. When transmission is suspected, investigation of potential donor-transmitted TB requires rapid communication among physicians, transplant centers, organ procurement organizations (OPOs), and public health authorities. PMID- 18385640 TI - Nonfatal maltreatment of infants--United States, October 2005-September 2006. AB - During October 2005-September 2006 (federal fiscal year 2006), approximately 905,000 U.S. children were victims of maltreatment that was substantiated by state and local child protective services (CPS) agencies. Approximately 19% of child maltreatment fatalities occurred among infants (i.e., persons aged <1 year), and homicide statistics suggest that fatality risk might be greatest in the first week of life. However, the risk for nonfatal maltreatment among infants has not been examined previously at the national level. To determine the extent of nonfatal infant maltreatment in the United States, CDC and the federal Administration for Children and Families (ACF) analyzed data collected in fiscal year 2006 (the most recent data available) from the National Child Abuse and Neglect Data System (NCANDS). This report summarizes the results of that analysis, which indicated that, in fiscal year 2006, a total of 91,278 infants aged <1 year (rate: 23.2 per 1,000 population) experienced nonfatal maltreatment, including 29,881 (32.7%) who were aged < or =1 week. Neglect was the maltreatment category cited for 68.5% of infants aged < or =1 week, but NCANDS data did not permit further characterization of the nature of this neglect. Developing effective measures to prevent maltreatment of infants aged < or =1 week will require more detailed characterization of neglect in this age group. PMID- 18385641 TI - Surveillance for community-associated Clostridium difficile--Connecticut, 2006. AB - Clostridium difficile is a well-known cause of hospital-acquired infectious diarrhea and is associated with increased health-care costs, prolonged hospitalizations, and increased patient morbidity. Previous antimicrobial use, especially use of clindamycin or ciprofloxacin, is the primary risk factor for development of C. difficile-associated diarrhea (CDAD) because it disrupts normal bowel flora and promotes C. difficile overgrowth. Historically, CDAD has been associated with elderly hospital in-patients or long-term--care facility (LTCF) residents. Since 2000, a strain of C. difficile that has been identified as North American pulsed-field type 1 (NAP1) and produces an extra toxin (binary toxin) and increased amounts of toxins A and B has caused increased morbidity and mortality among hospitalized patients. During 2005, related strains caused severe disease in generally healthy persons in the community at a rate of 7.6 cases per 100,000 population, suggesting that traditional risk factors for C. difficile might not always be factors in development of community-associated CDAD (CA CDAD). Cases of CA-CDAD are not nationally reportable, and population-based data at a statewide level have not been reported previously. In 2006, the Connecticut Department of Public Health (DPH) implemented a statewide surveillance system to assess the burden of CA-CDAD and to determine the descriptive epidemiology, trends, and risk factors for this disease. This report describes that surveillance system and summarizes results from the first year of surveillance. The findings indicated the presence of occasionally severe CDAD among healthy persons living in the community, including persons with no established risk factors for infection. Clinicians should consider a diagnosis of CA-CDAD in outpatients with severe diarrhea, even in the absence of established risk factors. In addition, continued surveillance is needed to determine trends in occurrence and whether more toxigenic strains are having an increasing impact in the community and in the hospital setting. PMID- 18385642 TI - Updated recommendation from the Advisory Committee on Immunization Practices (ACIP) for use of 7-valent pneumococcal conjugate vaccine (PCV7) in children aged 24-59 months who are not completely vaccinated. AB - This notice updates the recommendation for use of 7-valent pneumococcal conjugate vaccine (PCV7) among children aged 24-59 months who are either unvaccinated or who have a lapse in PCV7 administration. In February 2000, PCV7, marketed as Prevnar and manufactured by Wyeth Vaccines (Collegeville, Pennsylvania), was approved by the Food and Drug Administration for use in infants and young children. At that time, the Advisory Committee on Immunization Practices (ACIP) recommended that children aged 24-59 months who have certain underlying medical conditions or are immunocompromised receive PCV7. In addition, ACIP recommended that PCV7 be considered for all other children aged 24-59 months, with priority given to those who are American Indian/Alaska Native or of African-American descent, and to children who attend group day care centers. The recommendation also provided schedules for administering PCV7 to children aged 24-59 months who were either unvaccinated or who had a lapse in PCV7 administration; these schedules included 1) 1 dose of PCV7 for healthy children, and 2) 2 doses of PCV7 > or =2 months apart for children with certain chronic diseases or immunosuppressive conditions. PMID- 18385643 TI - [Coronary perfusion, necrosis area, pumping function of the myocardium, and cardiac electrical stability following revascularisation of infarction-affected myocardium]. AB - Analysing the literature data suggests that coronary angioplasty in acute myocardial infarction (AMI) fails to completely remove the focus of ischaemia, even in case of successful recanalization of the infarction-related artery. Inadequate post-angioplasty reperfusion of the infarction-stunned myocardium is mainly caused by the so-called <> phenomenon. Primary coronary angioplasty is capable of decreasing the size of the necrosis focus, being however incapable of completely removing myocardial infarction. Such a situation results from death of a considerable number of cardiomyocytes in the area of ischaemia, to have occurred long before coronary reperfusion is restored. An endovascular intervention in AMI may improve the impaired myocardial contractile performance and save the life of the patients sustaining the cardiogenic shock. However, coronary angioplasty does not preclude the development of cardiac insufficiency in all the patients suffering from acute myocardial infarction, which is a consequence of inadequate cardiac reperfusion and a result of necrotic death of a considerable segment of the heart muscle. Currently, there is no evidence regarding the effect of primary angioplasty on the risk of sudden cardiac death following AMI. An endovascular intervention on the infarction related artery cannot prevent the development of the postmyocardial-infarction cardiac remodelling, which is a consequence of incompletely restored coronary blood flow within the area of infarction. PMID- 18385644 TI - [Susceptibility to endothelium-dependent vasoactive agents in patients with coronary artery calcinosis]. AB - Coronary artery calcinosis was measured with ECG-gated multislice spiral CT in 68 patients, mean age 57.54+/-1.04 (M+/-m), including 47 men (69.1%) and 21 women (30.9%). Vascular reactivity of microcirculatory bed was assessed with laser Doppler flowmetry after acetylcholine and histamine administration. Coronary artery calcinosis correlated negatively with vascular susceptibility to acetylcholine (r=-0.380, p=0.002). This correlation did not depend on sex, age and concomitant drug therapy. Smoking was shown to affect significantly microvascular reactivity to histamine in persons free from coronary atherosclerosis. History of smoking determined the disturbances of vascular reactivity to histamine in patients with coronary artery calcinosis. PMID- 18385645 TI - [Haemorheological impairments following aortic surgery and methods of correction thereof]. AB - The article deals with the findings concerning alterations in rheological properties of the blood in patients who endured aortic surgery with and without artificial circulation. Also shown herein is the contribution of the plasmatic and erythrocytic components to the development of haemorheological disorders in the postoperative period. To this was added the assessment of the role plasmapheresis plays in correction of impaired blood rheological properties, depending upon the terms of its carrying out and the type of a surgical intervention performed. PMID- 18385646 TI - [Perftoran emulsion in complex therapy of critical lower limb ischemia]. AB - The paper presents comparative assessment of treatment outcomes in patients with critical lower limb ischemia treated with conventional therapy or combined with ozonized Perftoran. The study included 72 patients with Fontain-Pokrovsky III stage ischemia caused by impaired distal leg circulation. Thirty five patients received intra-arterial injections of ozonized Perftoran in addition to conventional "base" therapy. Emulsion of ozonized Perftoran was shown to have beneficial effect on blood coagulation, blood gases, parameters of systemic hemodynamics, microcirculation and other treatment outcome measures. PMID- 18385647 TI - [Peculiarities of ultrasonographic anatomy of the small saphenous vein]. AB - The authors studied the prevalence and peculiarities of lesions of the small saphenous vein (SSV), having performed ultrasonographic mapping of this vessel on 223 lower limbs. We revealed three major variants of localization of the SSV's terminal portion. In 36.3 % of cases, the SSV merged with the popliteal vein in the zone from the projection of the knee-join line and not above 7 cm from this level (type I). In 29.6% of cases, the SSV flowed into the femoral vein at a height of 8-20 cm from the knee-joint line (type II). In 31,3 % of cases, the SSV's terminal portion had no direct connection with the deep veins. In the upper third of the femur, the vessel flowed into the confluents of the large saphenous vein, or into the system of the gluteal veins (type III) The largest by the calibre are type I veins. Valvular insufficiency of the SSV was revealed in 16 (7.2 %) cases. The majority of the veins with incompetent valves (12) belonged to type I In the group of 143 veins with the diameter of not more than 0.4 cm at the level of the knee-joint fissure in the standing position, no pathological venous reflux was registered. We also examined the extent of the reflux along the SSV. Only in one case blood shunt to the lower third of the crus was registered. A combination of venous insufficiency of the SSV and the large saphenous vein on the same extremity was revealed only in 2 cases (1.6%). The obtained results give occasion to reconsider certain diagnostic and therapeutic approaches in varicose disease in the SSV's basin. PMID- 18385648 TI - [The role of laser doppler flowmetry in the assessment of microcirculatory disturbances at angiodysplasias]. AB - The paper presents the data on the use of laser Doppler flowmetry (LDF) for microcirculation assessment in patients with lower limb angiodysplasias. Preliminary results suggest that LDF can be successfully used both for the quantification of microcirculatory disturbances and for the diagnosis of arterio venous micro fistulas. LDF was used as a monitoring tool for compressive therapy effectiveness in patients with arterio-venous and venous forms of lower limb angiodysplasias. Compression stockings Relaxsan III class were shown to improve microcirculation in this patient population after 1 month wearing. PMID- 18385649 TI - [CT perfusion for assessment of brain stem ischemic lesions]. AB - Modern neurovisualization modalities - CT and MRI with cerebral circulation assessment was used for diagnosis of cerebrovascular disturbances in patients admitted to the Emergency Care Hospital of Ufa. CT and MRI perfusion methods appeared to be highly effective both in diagnosis and treatment efficacy monitoring of acute stroke. PMID- 18385650 TI - Endovascular treatment of a renal artery stenosis: techniques, indications, results. Role of embolic protection devices (part 4). PMID- 18385651 TI - [Endovascular treatment of major vascular aneurysms: successful experience]. AB - Major vascular aneurysm is a severe pathology with poor prognosis. Due to wide variations of vascular anatomy, an endovascular intervention may require intraoperative adjustment of procedural strategy. The paper presents results of successive stent-graft implantations in different anatomical situations. PMID- 18385652 TI - [Endovascular microembolization of the posterior auricular artery in comprehensive surgical management of capillary angioma of the floor of the auricle]. AB - The article deals with the description of a rare clinical case wherein a 44-year old female patient presenting with bleeding capillary angioma of the left auricle obstructing the auditory passage was fist subjected to endovascular microembolization of the posterior auricular artery by means of polyvinyl alcohol (PVA-500), followed by a second stage of virtually bloodlessly surgical removal of the tumour carried out five days thereafter. This case report once again points to feasibility of combined, i. e. endovascular and conventional surgical management of capillary angiomas. Preoperative microembolization of a bleeding haemangioma arrests haemorrhage from the tumour, resulting in the decreased size thereof, as well as leading to circumscribing the tumour from healthy tissue. Attention is paid to the fact that embolization of the auricular artery should be performed only after a thorough angiographic evaluation of the pattern of the blood flow in the tumour and adjacent vessels by means of the technique of selective catheterization and using only the officinal emboli. PMID- 18385653 TI - [What was interesting at the World Congress of the International Union of Phlebology. Kyoto, 18-20 June, 2007]. AB - Asian Chapter Meeting of the International Union of Phleboiogy (UIP) was held in Kyoto (Japan), 18?-?20 June, 2007. Participants of this important forum have discussed some actual problems concerning epidemiology, pathogenesis, diagnosis and treatment of venous diseases. PMID- 18385654 TI - [Syndrome of lower limb volume enlargement in varicosity: causes and medical approaches]. AB - This paper provides the results of examination of 413 patients aged 15 to 79 years (mean age 43.2 years) with different stages of CVI seen from 1995 to 2005. A complex of methods was employed: the clinical nmltimodality examination, multilevel volumetric monitoring before and after daily orthostatic loading, occlusion plethysmography, multilevel ultrasound duplex scanning of the venous system arid subcutis as well as radiography. Only in the third of cases (n=149, 36.0%) was primarily of distal character attended by lesion of the foot and leg, with clear daily dynamics and dependence on the daily orthostatic loading-eaema proper. Two types of edema were distinguished: 1)with pronounced daily dynamics and return to the initial parameters in the morning hours (131 patients) and 2) with moderate daily dynamics of edema and the lack of return to the initial parameters in the morning hours (18 patients). Such difference stems from the type of coexistent lymphatic insufficiency (with a high and low drop). However, only the first type is initially determined by venous insufficiency. The majorityof this group patients were found to suffer thrombosis of the ti-bial veins in the presence of varicosity running a prolonged course. Hinetyone patients (22.0%) had a proximally oriented enlargement, with pronounced orthostasodependerice, of the muscle segments of the limb because of orthostasodependent phlebopaxhy. In the remaining 173 (41.9%) patients examined, vascular eaema was coupled with non-vascular causes of limb size enlargement (arthrosis of leg joints, lipoedema) or the latter ones were isblated. The aim of the present study was to analyze the causes of the syndrome of limb size enlargement in varicosity with the purpose of upgrading the efficacy and trend of the treatment. PMID- 18385655 TI - [Acerbin for treatment of indurative cellulite in patients with venous trophic ulcers]. AB - Acute indurative cellulite can impair regeneration processes in patients with venous trophic ulcers. Wide range of antimicrobial agents is used for its management, while their role and place remain disputable. The choice of adequate local therapy plays a key role in total success. Conventional topical drugs and anticeptics in most cases appear to be ineffective or, besides antimicrobial action, can inhibit reparative processes. Combined formulation Acerbin exhibits not only antimicrobial properties, but stimulates granulation and epithelization, as well, and resolves inflammation in adjacent to ulcer tissues. That is why it can serve a beneficial alternative to conventional topical drugs. PMID- 18385656 TI - [Treatment of inferior vena cava thrombosis. How to avoid errors?]. AB - Unsatisfactory results of acute thrombosis treatment in inferior vena cava system are attributed to inadequate diagnosis, poor compliance with approved clinical practice guidelines and secondary preventive measures, as well as to excessive adherence to surgical methods of pulmonary embolism prophylaxis. Diagnostic strategy, which combines compressive duplex scanning and D-dimer test, can improve diagnosis and reduce its cost. Various molecular weight heparins and vitamin K antagonists are still the main means of venous thrombosis therapy. Appropriate medical treatment in many cases helps to avoid surgical interventions for pulmonary embolism prophylaxis and to reduce the rate of recurrent thromboses. PMID- 18385657 TI - [Indications for using an intraluminal temporary bypass in operations on carotid arteries]. AB - The review is dedicated to the most commonly used method of protecting the brain in carotid endarterectomy, i. e. using an intraluminal temporary bypass (ITB), also containing the information concerning the constructions of ITBs, various points of view regarding practical application thereof, as well as the methods of monitoring the functions of the brain in order to solve the problem concerning feasibility of using an ITB. PMID- 18385658 TI - Carotid artery grafting for restenosis after carotid endarterectomy. AB - The paper analyzes outcomes of 12 internal carotid artery (ICA) graftings for repeat stenosis after carotid endarterectomy (CEA) in 11 patients. In 6 cases restenosis occurred after eversion CEA, in 5 - after conventional patch CEA, and in 1 case - after ICA patch angioplasty for post-CEA restenosis. Critical restenosis >75% in reconstruction area was an indication for surgical intervention. There was 1 case of graft thrombosis in early postoperative period with hemispherical stroke and lethal outcome. Eight patients were examined in late postoperative period. Mean follow-up was 34?+/-?8.44 months (from 16 to 48 months). No cerebrovascular disturbances in operated ICA feeding area were detected. In 6 patients graft patency was good, while 2 smoking patients with diabetes mellitus and hyperlipidemia demonstrated repeat restenosis of proximal anastomosis >70%. PMID- 18385659 TI - [Simultaneous interventions on the ascending portion, arch of the aorta and cardiac valves in patients with Marfan's syndrome]. AB - Simultaneous surgical interventions on the aorta and valvular system of the heart were performed in four patients presenting with aortic dissections and aneurysms conditioned by Marfan's syndrome. The following reconstructive operations were carried out: 1) prosthetic repair of the aortic valve and the ascending portion of the aorta by means of a valve-containing conduit with replantation of the openings of the coronary arteries into the side of the prosthesis according to the Benthall - De Bono technique, annuloplasty of the tricuspid valve according to the De Vega technique, valvuloplasty of the mitral valve by the Alferi technique; 2) grafting of the aortic valve and the ascending portion of the aorta by means of a valve-containing conduit with replantation of the openings of the coronary arteries according to the Kabrol's technique, plasty of the tricuspid valve by the De Vega technique; 3) prosthetic repair of the aortic arch with distal wedge-like excision of the membrane of the dissection and directing the blood flow along the both channels, plasty of the mitral valve, plasty of the aortic valve and the ascending portion of the aorta with a valve-containing conduit, accompanied by replantation of the openings of the coronary arteries into the side of the graft according to the Benthall - De Bono technique; (4) plasty of the mitral valve with a disk graft through the fibrous ring of the aortic valve, prosthetic repair of the aortic valve and the ascending portion of the aorta with a valve-containing conduit, accompanied by replantation of the openings of the coronary arteries into the side according to the Benthall-De Bono technique. PMID- 18385660 TI - [Transformation of bioprosthesis-artery anastomotic area after blood exposure: the effect of suture material]. AB - Vascular graft implantation is accompanied with tissue reaction which is determined partially by the nature of suture material. In experiments in vitro edges of arterial diepoxid-preserved bioprostheses were sutured with Prolene filament, nickelide-titanium wire (TiNi) or absorbable polydioxanone monofilament (PDS). Morphological changes in anastomotic area after blood exposure were assessed with scanning electronic microscope. Protein deposits in anastomotic area were detected 5 min after blood exposure, being more loose and bulky around Prolene sutures. The protein layer became more compact 120 min since blood exposure. Most homogeneous protein sorption was over TiNi-sutured anastomoses. Suture material affected blood cells as well - some red cells have transformed into echinocytes. Heparin-modified samples demonstrated less negative impact of suture material in anastomotic area. The worst results after 120 min of blood exposure were obtained for PDS filaments. PMID- 18385661 TI - Local repair of distal thoracal aortic dissections (Locus minoris resistencia). AB - The paper presents the method of local repair of distal aortic dissections. Local aortic grafting for surgical correction of type B dissecting aortic aneurysms helped to decrease hospital mortality up to 15.4%, the rate of paraparesis and multiorgan failure - up to 11.5%. PMID- 18385662 TI - [Hyperplasia of the intima as a cause of restenosis following reconstructive and endovascular operations on lower limb arteries]. AB - The authors herein analyse the role of intimal hyperplasia in the development of restenosis as a late complication in reconstructive surgery of arteries in patients suffering from atherosclerosis obliterans of the arteries of the lower extremities. PMID- 18385663 TI - Outcomes of conventional surgery and combined with endovascular intervention for acute thromboses of atherosclerotically arteries comparative study. AB - The study was aimed at early outcome retrospective analysis of conventional surgery (CS) for acute arterial obstruction (AAO) of lower limb arteries with atherosclerotic lesions and of simultaneous combined treatment (SCT) - endovascular intervention undertaken during open surgery. MATERIALS AND METHODS: 166 histories of patients with AAO and atherosclerotic lesions of lower limb arteries, treated during 1998-2005. Conventional surgical therapy was used in 72 cases and comprised open and indirect thrombectomy with endarterectomy and/or femoral arterioplasty if indicated. In 94 eases open surgery was combined with pre- or intraoperative angiography and endovascular balloon angioplasty. RESULTS: restoration of blood flow during primary intervention was achieved in 58.3% of patients after CS and in 72.3% of patients after SCT; the rate of amputations was 31.9% and 18.1%, respectively, mortality - 16.6% and 6.3%, respectively, favorable outcome at the discharge - 62.5% and 79.9%. CONCLUSION: endovascular angioplasty of atherosclerotic stenoses during open surgery for acute thromboses of lower limb arteries has distinct benefits over conventional surgical treatment. PMID- 18385664 TI - Diabetes and radiocontrast media increase endothelin converting enzyme-1 in the kidney. AB - Plasma endothelin-1 levels rise in diabetes and after exposure to contrast media suggesting a role in progressive diabetic and acute radiocontrast nephropathies. Here we studied individual and combined effects of streptozotocin-induced diabetes and contrast media on renal endothelin converting enzyme-1 levels in the rat. In vivo, medullary (but not cortical) endothelin converting enzyme protein gradually increased 4 to 5-fold following the induction of diabetes or after the administration of contrast media but rose 15-fold when diabetic rats were given contrast media. Changes in mRNA expression paralleled those of the protein. Immunohistochemistry confirmed that increased tubular and endothelial cell endothelin converting enzyme-1 were most pronounced in the medulla. In vitro, endothelin-1 levels increased 3-fold following incubation of endothelial cells with media high in glucose or with contrast and 4-fold with their combination. Endothelin converting enzyme-1 protein and mRNA expression changed in a similar pattern while prepro endothelin-1 mRNA increased with each insult but not in an additive way. Our study shows that diabetes and contrast media up-regulate renal medullary endothelin converting enzyme-1 expression and synthesis. PMID- 18385665 TI - A mouse model for polycystic kidney disease through a somatic in-frame deletion in the 5' end of Pkd1. AB - Autosomal dominant polycystic kidney disease, a leading cause of end-stage renal disease in adults, is characterized by progressive focal cyst formation in the kidney. Embryonic lethality of Pkd1-targeted mice limits the use of these mice. Here we developed a floxed allele of Pkd1 exons 2-6. Global deletion mutants developed polyhydramnios, hydrops fetalis, polycystic kidney and pancreatic disease. Somatic Pkd1 inactivation in the kidney was achieved by crossing Pkd1(flox) mice with transgenic mice expressing Cre controlled by a gamma glutamyltranspeptidase promoter. These mutants developed cysts in both proximal and distal nephron segments and survived for about 4 weeks. Somatic loss of heterozygosity was shown in a reporter mouse strain to cause cystogenesis. Some cysts in young mice are positive for multiple tubular markers and a mesenchymal marker, suggesting a delay in tubular epithelial differentiation. A higher cell proliferation rate was observed in distal nephron segments probably accounting for the faster growth rate of distal cysts. Although we observed an overall increase in apoptosis in cystic kidneys, there was no difference between proximal or distal nephron segments. We also found increased cyclic AMP, aquaporin 2 and vasopressin type 2 receptor mRNA levels, and apical membrane translocation of aquaporin 2 in cystic kidneys, all of which may contribute to the differential cyst growth rate observed. The accelerated polycystic kidney phenotype of these mice provides an excellent model for studying molecular pathways of cystogenesis and to test therapeutic strategies. PMID- 18385666 TI - Failure to phosphorylate AKT in podocytes from mice with early diabetic nephropathy promotes cell death. AB - Loss of podocytes by apoptosis characterizes the early stages of diabetic nephropathy. To examine its mechanism we studied glomeruli and podocytes isolated from db/db mice with early diabetic nephropathy and albuminuria. Phosphorylation of AKT (protein kinase B, a key survival protein) was found to be lower in the glomeruli of 12 week old db/db compared to db/+ mice. In vitro, insulin phosphorylated AKT solely in podocytes from db/+ mice. Serum deprivation and exposure to tumor necrosis factor-alpha significantly compromised cell viability in podocytes from db/db but not from db/+ mice, and this was associated with a significant decrease in AKT phosphorylation. Inhibition of AKT was necessary to achieve the same degree of cell death in db/+ podocytes. Our study shows that podocyte inability to respond to insulin and susceptibility to cell death may partially account for the decreased podocyte number seen in early diabetic nephropathy. PMID- 18385667 TI - Improvement of cast nephropathy with plasma exchange depends on the diagnosis and on reduction of serum free light chains. AB - Cast nephropathy is the most common cause of renal disease in multiple myeloma, however, treatment with plasma exchange remains controversial even after 3 randomized controlled studies. We sought to determine the importance of diagnostic confirmation and goal directed therapy in the treatment of cast nephropathy in forty patients with confirmed multiple myeloma and renal failure who underwent plasma exchange. A positive renal response was defined as a decrease by half in the presenting serum creatinine and dialysis independence. No baseline differences were noted between eventual renal responders and non responders. Three quarters of the patients with biopsy proven cast nephropathy resolved their renal disease when the free light chains present in the serum were reduced by half or more but there was no significant response when the reduction was less. The median time to a response was about 2 months. In patients without cast nephropathy, renal recovery occurred despite reductions in free light chain levels of the serum. No association was found between free light chains in the serum, urinary monoclonal proteins, overall proteinuria and cast nephropathy. We found that the relationship between renal recovery and free light chain reduction was present only in patients with biopsy proven cast nephropathy showing the importance of extracorporeal light chain removal in this disease. PMID- 18385668 TI - The risk of acute renal failure in patients with chronic kidney disease. AB - Few studies have defined how the risk of hospital-acquired acute renal failure varies with the level of estimated glomerular filtration rate (GFR). It is also not clear whether common factors such as diabetes mellitus, hypertension and proteinuria increase the risk of nosocomial acute renal failure independent of GFR. To determine this we compared 1,746 hospitalized adult members of Kaiser Permanente Northern California who developed dialysis-requiring acute renal failure with 600,820 hospitalized members who did not. Patient GFR was estimated from the most recent outpatient serum creatinine measurement prior to admission. The adjusted odds ratios were significantly and progressively elevated from 1.95 to 40.07 for stage 3 through stage 5 patients (not yet on maintenance dialysis) compared to patients with estimated GFR in the stage 1 and 2 range. Similar associations were seen after controlling for inpatient risk factors. Pre admission baseline diabetes mellitus, diagnosed hypertension and known proteinuria were also independent risk factors for acute kidney failure. Our study shows that the propensity to develop in-hospital acute kidney failure is another complication of chronic kidney disease whose risk markedly increases even in the upper half of stage 3 estimated GFR. Several common risk factors for chronic kidney disease also increase the peril of nosocomial acute kidney failure. PMID- 18385670 TI - Exposure information in environmental health research: current opportunities and future directions for particulate matter, ozone, and toxic air pollutants. AB - Understanding and quantifying outdoor and indoor sources of human exposure are essential but often not adequately addressed in health effect studies for air pollution. Air pollution epidemiology, risk assessment, health tracking, and accountability assessments are examples of health effect studies that require but often lack adequate exposure information. Recent advances in exposure modeling along with better information on time-activity and exposure factor data provide us with unique opportunities to improve the assignment of exposures for both future and ongoing studies linking air pollution to health impacts. In September 2006, scientists from the US Environmental Protection Agency and the Centers for Disease Control and Prevention along with scientists from the academic community and state health departments convened a symposium on air pollution exposure and health to identify, evaluate, and improve current approaches for linking air pollution exposures to disease. This manuscript presents the key issues, challenges, and recommendations identified by the exposure working group, who used case studies of particulate matter, ozone, and toxic air-pollutant exposure to evaluate health effects for air pollution. One of the overarching lessons of this workshop is that obtaining better exposure information for these different health effect studies requires both goal setting for what is needed and mapping out the transition pathway from current capabilities for meeting these goals. Meeting our long-term goals requires definition of incremental steps that provide useful information for the interim and move us toward our long-term goals. Another overarching theme among the three different pollutants and the different health study approaches is the need for integration among alternate exposure assessment approaches. For example, different groups may advocate exposure indicators, biomonitoring, mapping methods (GIS), modeling, environmental media monitoring, and/or personal exposure modeling. However, emerging research reveals that the greatest progress comes from integration among two or more of these efforts. PMID- 18385669 TI - Calcium-independent and 1,25(OH)2D3-dependent regulation of the renin-angiotensin system in 1alpha-hydroxylase knockout mice. AB - To determine whether the cardiovascular effect of 1,25(OH)(2)D is dependent on calcium and/or phosphorus, mice with targeted deletion of the 25(OH)D 1alpha hydroxylase and their wild-type littermates were fed a normal diet or a diet to rescue the ambient serum calcium and phosphorus levels. Mice on the normal diet were treated daily with vehicle or 1,25(OH)(2)D(3) while mice on the rescue diet received vehicle, captopril or losartan. After four weeks the vehicle-treated knockout mice developed hypertension, cardiac hypertrophy and impaired cardiac function along with an up-regulation of the renin-angiotensin system in both renal and cardiac tissues. Although the serum calcium and phosphorus levels were normalized in knockout mice on the rescue diet, abnormalities in blood pressure, cardiac structure-function and the renin-angiotensin system remained. In contrast, 1,25(OH)(2)D(3) not only normalized serum calcium and phosphorus levels but also normalized blood pressure, cardiac structure-function and the renin angiotensin system. Treatment of the knockout mice with either captopril or losartan normalized blood pressure and cardiac structure and function although renin expression remained elevated. This study shows that 1,25(OH)2D plays a protective role in the cardiovascular system by repressing the renin-angiotensin system independent of extracellular calcium or phosphorus. PMID- 18385671 TI - Developing probability distributions for transfer efficiencies for dermal exposure. AB - Many dermal exposure models use stochastic techniques to sample parameter distributions derived from experimental data to more accurately represent variability and uncertainty. Transfer efficiencies represent the fraction of a surface contaminant transferred from the surface to the skin during a contact event. Although an important parameter for assessing dermal exposure, examination of the literature confirms that no single study is large enough to provide a basis for a transfer efficiency distribution for use in stochastic dermal exposure models. It is therefore necessary to combine data sets from multiple studies to achieve the largest data set possible for distribution analysis. A literature review was conducted to identify publications reporting transfer efficiencies. Data sets were compared using the Kruskal-Wallis test to determine whether they arise from the same distribution. Combined data were evaluated for several theoretical distributions using the Kolmogorov-Smirnov and chi(2) goodness-of-fit tests. Our literature review identified 35 studies comprising 25 different sampling methods, 25 chemicals, and 10 surface types. Distributions were developed for three different chemicals (chlorpyrifos, pyrethrin I, and piperonyl butoxide) on three different surface types (carpet, vinyl, and foil). Only the lognormal distribution was consistently accepted for each chemical and surface combination. Fitted distributions were significantly different (Kruskal Wallis test; P<0.001) across chemicals and surface types. In future studies, increased effort should be placed on developing large studies, which more accurately represent transfer to human skin from surfaces, and on developing a normative transfer efficiency measure so that data from different methodologies can be compared. PMID- 18385672 TI - Discrete fixed-resolution representations in visual working memory. AB - Limits on the storage capacity of working memory significantly affect cognitive abilities in a wide range of domains, but the nature of these capacity limits has been elusive. Some researchers have proposed that working memory stores a limited set of discrete, fixed-resolution representations, whereas others have proposed that working memory consists of a pool of resources that can be allocated flexibly to provide either a small number of high-resolution representations or a large number of low-resolution representations. Here we resolve this controversy by providing independent measures of capacity and resolution. We show that, when presented with more than a few simple objects, human observers store a high resolution representation of a subset of the objects and retain no information about the others. Memory resolution varied over a narrow range that cannot be explained in terms of a general resource pool but can be well explained by a small set of discrete, fixed-resolution representations. PMID- 18385674 TI - Imaging of Rab5 activity identifies essential regulators for phagosome maturation. AB - Efficient phagocytosis of apoptotic cells is crucial for tissue homeostasis and the immune response. Rab5 is known as a key regulator of the early endocytic pathway and we have recently shown that Rab5 is also implicated in apoptotic cell engulfment; however, the precise spatio-temporal dynamics of Rab5 activity remain unknown. Here, using a newly developed fluorescence resonance energy transfer biosensor, we describe a change in Rab5 activity during the engulfment of apoptotic thymocytes. Rab5 activity on phagosome membranes began to increase on disassembly of the actin coat encapsulating phagosomes. Rab5 activation was either continuous or repetitive for up to 10 min, but it ended before the collapse of engulfed apoptotic cells. Expression of a dominant-negative mutant of Rab5 delayed this collapse of apoptotic thymocytes, showing a role for Rab5 in phagosome maturation. Disruption of microtubules with nocodazole inhibited Rab5 activation on the phagosome membrane without perturbing the engulfment of apoptotic cells. Furthermore, we found that Gapex-5 is the guanine nucleotide exchange factor essential for Rab5 activation during the engulfment of apoptotic cells. Gapex-5 was bound to a microtubule-tip-associating protein, EB1, whose depletion inhibited Rab5 activation during phagocytosis. We therefore propose a mechanistic model in which the recruitment of Gapex-5 to phagosomes through the microtubule network induces the transient Rab5 activation. PMID- 18385673 TI - Genetic variation in human NPY expression affects stress response and emotion. AB - Understanding inter-individual differences in stress response requires the explanation of genetic influences at multiple phenotypic levels, including complex behaviours and the metabolic responses of brain regions to emotional stimuli. Neuropeptide Y (NPY) is anxiolytic and its release is induced by stress. NPY is abundantly expressed in regions of the limbic system that are implicated in arousal and in the assignment of emotional valences to stimuli and memories. Here we show that haplotype-driven NPY expression predicts brain responses to emotional and stress challenges and also inversely correlates with trait anxiety. NPY haplotypes predicted levels of NPY messenger RNA in post-mortem brain and lymphoblasts, and levels of plasma NPY. Lower haplotype-driven NPY expression predicted higher emotion-induced activation of the amygdala, as well as diminished resiliency as assessed by pain/stress-induced activations of endogenous opioid neurotransmission in various brain regions. A single nucleotide polymorphism (SNP rs16147) located in the promoter region alters NPY expression in vitro and seems to account for more than half of the variation in expression in vivo. These convergent findings are consistent with the function of NPY as an anxiolytic peptide and help to explain inter-individual variation in resiliency to stress, a risk factor for many diseases. PMID- 18385677 TI - Multidisciplinary aspect of spinal cord medicine. Introduction. PMID- 18385675 TI - A deadenylation negative feedback mechanism governs meiotic metaphase arrest. AB - In vertebrate oocytes, meiotic progression is driven by the sequential translational activation of maternal messenger RNAs stored in the cytoplasm. This activation is mainly induced by the cytoplasmic elongation of their poly(A) tails, which is mediated by the cytoplasmic polyadenylation element (CPE) present in their 3' untranslated regions. In Xenopus oocytes, sequential phase-specific translation of CPE-regulated mRNAs is required to activate the maturation promoting factor, which in turn mediates entry into the two consecutive meiotic metaphases (MI and MII). Here we report a genome-wide functional screening to identify previously unknown mRNAs cytoplasmically polyadenylated at meiotic phase transitions. A significant fraction of transcripts containing, in addition to CPEs, (A + U)-rich element (ARE) sequences (characteristic of mRNAs regulated by deadenylation) were identified. Among these is the mRNA encoding C3H-4, an ARE binding protein that we find to accumulate in MI and the ablation of which induces meiotic arrest. Our results suggest that C3H-4 recruits the CCR4 deadenylase complex to ARE-containing mRNAs and this, in turn, causes shortening of poly(A) tails. We also show that the opposing activities of the CPEs and the AREs define the precise activation times of the mRNAs encoding the anaphase promoting complex inhibitors Emi1 and Emi2 during distinct phases of the meiotic cycle. Taken together, our results show that an 'early' wave of cytoplasmic polyadenylation activates a negative feedback loop by activating the synthesis of C3H-4, which in turn would recruit the deadenylase complex to mRNAs containing both CPEs and AREs. This negative feedback loop is required to exit from metaphase into interkinesis and for meiotic progression. PMID- 18385676 TI - Genome-wide association scan of tag SNPs identifies a susceptibility locus for lung cancer at 15q25.1. AB - To identify risk variants for lung cancer, we conducted a multistage genome-wide association study. In the discovery phase, we analyzed 315,450 tagging SNPs in 1,154 current and former (ever) smoking cases of European ancestry and 1,137 frequency-matched, ever-smoking controls from Houston, Texas. For replication, we evaluated the ten SNPs most significantly associated with lung cancer in an additional 711 cases and 632 controls from Texas and 2,013 cases and 3,062 controls from the UK. Two SNPs, rs1051730 and rs8034191, mapping to a region of strong linkage disequilibrium within 15q25.1 containing PSMA4 and the nicotinic acetylcholine receptor subunit genes CHRNA3 and CHRNA5, were significantly associated with risk in both replication sets. Combined analysis yielded odds ratios of 1.32 (P < 1 x 10(-17)) for both SNPs. Haplotype analysis was consistent with there being a single risk variant in this region. We conclude that variation in a region of 15q25.1 containing nicotinic acetylcholine receptors genes contributes to lung cancer risk. PMID- 18385678 TI - Penile prosthesis implantation: past, present and future. AB - Penile prosthesis implantation is the oldest effective treatment for erectile dysfunction. This review examines the past, present and future of penile prosthesis implantation. Advances in prosthetic design and implantation techniques have resulted today in devices that produce nearly normal flaccid and erect states, and have remarkable freedom from mechanical failure. The future of prosthetic design holds promises for even more improvements. PMID- 18385679 TI - Discovery of NO in the penis. AB - In 1989, the recognition by our group at UCLA that nitric oxide was indeed the chemical mediator of penile erection turned out to have major influences on our society and on some of those who participated in this research. Not only did it ultimately lead to the development of drugs like sildenafil, vardenafil and tadalafil, it launched new businesses, it gave insight into the way the vascular system functioned, it changed the way many physicians, particularly urologists, practiced sexual medicine and by bringing the topic of erectile dysfunction out of the closet, talk of sexual issues became mainstream. For one of us, a Nobel Prize lay in the future. This review, which comes one decade following the introduction of Viagra into the marketplace, chronicles the actual events involved in the discovery. PMID- 18385680 TI - Regulation of interferon production and innate antiviral immunity through translational control of IRF-7. PMID- 18385681 TI - Ethnic differences in infant mortality by cause of death. AB - OBJECTIVE: The purpose of this study was to examine ethnic differences among non Hispanic black and white births in the distribution of maternal risk factors of infant mortality across specific causes of death. STUDY DESIGN: The data were obtained from the North Carolina linked birth/infant death files (1989 to 1997). Logistic regression models were built to assess the risk distribution of demographic, behavioral and health related variables in relation to causes of infant death, which included congenital anomalies, short gestation/low birth weight (LBW), sudden infant death syndrome, infections and obstetric conditions. RESULT: Infants born to black women had the highest rates for all causes of mortality compared to those born to white women. Having at least one prior live birth now dead was associated with congenital anomalies, obstetric conditions and short gestation/LBW related deaths in both ethnic groups. Deaths caused by infections were more likely to occur among white young (<20) women. White women enrolled in Medicaid had an increased risk of infant deaths due to short gestation/LBW when compared to those with no Medicaid, while young black mothers (<20 years old) were less likely to experience an infant death due to short gestation/LBW and obstetric conditions. CONCLUSION: This study provides evidence that maternal sociodemographic risk factors somewhat vary by infant cause of death and ethnicity. This suggests that race-specific approaches may be necessary to reduce infant mortality rates. The differences, however, in the risk distribution of factors across the two ethnic groups were limited indicating that the heterogeneity in the mortality rates may be due to unmeasured factors. PMID- 18385682 TI - The challenge of cancer control in Africa. AB - While the world is focused on controlling the spread of diseases such as HIV and malaria in the developing world, another approaching epidemic has been largely overlooked. The World Heath Organization predicts that there will be 16 million new cancer cases per year in 2020 and 70% of these will be in the developing world. Many of these cancers are preventable, or treatable when detected early enough. Establishing effective, affordable and workable cancer control plans in African countries is one step in the right direction toward limiting this epidemic. PMID- 18385683 TI - Fusion of tumour cells with bone marrow-derived cells: a unifying explanation for metastasis. AB - The causes of metastasis remain elusive despite vast information on cancer cells. We posit that cancer cell fusion with macrophages or other migratory bone marrow derived cells (BMDCs) provides an explanation. BMDC-tumour hybrids have been detected in numerous animal models and recently in human cancer. Molecular studies indicate that gene expression in such hybrids reflects a metastatic phenotype. Should BMDC-tumour fusion be found to underlie invasion and metastasis in human cancer, new approaches for therapy would surely follow. PMID- 18385688 TI - Double vision. PMID- 18385684 TI - Getting to the stem of chronic myeloid leukaemia. AB - Tyrosine kinase inhibitor (TKI) therapy for chronic myeloid leukaemia (CML) is the consummate success story for targeted therapy, yet relapse is a nearly inevitable consequence of cessation or interruption of therapy. Primitive TKI refractory CML stem cells are the likely source of these relapses, as they provide sanctuary for the Philadelphia chromosome. In advanced disease, their progressively anaplastic progeny ultimately maintain CML independently of the CML haematopoietic stem cell (HSC). Interestingly, there are at least two distinct cell types capable of self-renewal in different phases of CML: first, a primitive HSC with BCR-ABL mutation, which maintains the more indolent chronic-phase disease and, second, a coexisting mutated progenitor cell which acquires stem cell characteristics responsible for rapid cell expansion in advanced disease. PMID- 18385689 TI - Broken promises. PMID- 18385690 TI - Mismanaged measures. PMID- 18385693 TI - Carbon-trading market has uncertain future. PMID- 18385694 TI - Are the IPCC scenarios 'unachievable'? PMID- 18385695 TI - Drug markers questioned. PMID- 18385697 TI - Easy ways to other Earths. PMID- 18385696 TI - EPA feels heat over flame retardant. PMID- 18385707 TI - Hazy reasoning behind clean air. PMID- 18385708 TI - Architecture: architects of a low-energy future. PMID- 18385709 TI - Anatomy: truly gross anatomy. PMID- 18385710 TI - Portugal: navigating new waters. PMID- 18385711 TI - Give south Indian authors their true names. PMID- 18385712 TI - Spanish funds will finance Bilbao's spallation bid. PMID- 18385713 TI - Comparing the legacies of Gauss, Pasteur and Darwin. PMID- 18385715 TI - Dangerous assumptions. PMID- 18385720 TI - Genomics: when the smoke clears ... PMID- 18385721 TI - Extrasolar planets: with a coarse-tooth comb. PMID- 18385722 TI - Geophysics: humming a different tune. PMID- 18385723 TI - Drug resistance: the fight against fungi. PMID- 18385724 TI - Condensed-matter physics: opposite of a superconductor. PMID- 18385725 TI - RNA interference: generic block on angiogenesis. PMID- 18385726 TI - Obituary: Arthur C. Clarke (1917-2008). PMID- 18385727 TI - Molecular cancer diagnostics. PMID- 18385728 TI - The cancer biomarker problem. AB - Genomic technologies offer the promise of a comprehensive understanding of cancer. These technologies are being used to characterize tumours at the molecular level, and several clinical successes have shown that such information can guide the design of drugs targeted to a relevant molecule. One of the main barriers to further progress is identifying the biological indicators, or biomarkers, of cancer that predict who will benefit from a particular targeted therapy. PMID- 18385730 TI - Enabling personalized cancer medicine through analysis of gene-expression patterns. AB - Therapies for patients with cancer have changed gradually over the past decade, moving away from the administration of broadly acting cytotoxic drugs towards the use of more-specific therapies that are targeted to each tumour. To facilitate this shift, tests need to be developed to identify those individuals who require therapy and those who are most likely to benefit from certain therapies. In particular, tests that predict the clinical outcome for patients on the basis of the genes expressed by their tumours are likely to increasingly affect patient management, heralding a new era of personalized medicine. PMID- 18385729 TI - Translating insights from the cancer genome into clinical practice. AB - Cancer cells have diverse biological capabilities that are conferred by numerous genetic aberrations and epigenetic modifications. Today's powerful technologies are enabling these changes to the genome to be catalogued in detail. Tomorrow is likely to bring a complete atlas of the reversible and irreversible alterations that occur in individual cancers. The challenge now is to work out which molecular abnormalities contribute to cancer and which are simply 'noise' at the genomic and epigenomic levels. Distinguishing between these will aid in understanding how the aberrations in a cancer cell collaborate to drive pathophysiology. Past successes in converting information from genomic discoveries into clinical tools provide valuable lessons to guide the translation of emerging insights from the genome into clinical end points that can affect the practice of cancer medicine. PMID- 18385731 TI - Mining the plasma proteome for cancer biomarkers. AB - Systematic searches for plasma proteins that are biological indicators, or biomarkers, for cancer are underway. The difficulties caused by the complexity of biological-fluid proteomes and tissue proteomes (which contribute proteins to plasma) and by the extensive heterogeneity among diseases, subjects and levels of sample procurement are gradually being overcome. This is being achieved through rigorous experimental design and in-depth quantitative studies. The expected outcome is the development of panels of biomarkers that will allow early detection of cancer and prediction of the probable response to therapy. Achieving these objectives requires high-quality specimens with well-matched controls, reagent resources, and an efficient process to confirm discoveries through independent validation studies. PMID- 18385733 TI - A nuclear receptor-like pathway regulating multidrug resistance in fungi. AB - Multidrug resistance (MDR) is a serious complication during treatment of opportunistic fungal infections that frequently afflict immunocompromised individuals, such as transplant recipients and cancer patients undergoing cytotoxic chemotherapy. Improved knowledge of the molecular pathways controlling MDR in pathogenic fungi should facilitate the development of novel therapies to combat these intransigent infections. MDR is often caused by upregulation of drug efflux pumps by members of the fungal zinc-cluster transcription-factor family (for example Pdr1p orthologues). However, the molecular mechanisms are poorly understood. Here we show that Pdr1p family members in Saccharomyces cerevisiae and the human pathogen Candida glabrata directly bind to structurally diverse drugs and xenobiotics, resulting in stimulated expression of drug efflux pumps and induction of MDR. Notably, this is mechanistically similar to regulation of MDR in vertebrates by the PXR nuclear receptor, revealing an unexpected functional analogy of fungal and metazoan regulators of MDR. We have also uncovered a critical and specific role of the Gal11p/MED15 subunit of the Mediator co-activator and its activator-targeted KIX domain in antifungal/xenobiotic-dependent regulation of MDR. This detailed mechanistic understanding of a fungal nuclear receptor-like gene regulatory pathway provides novel therapeutic targets for the treatment of multidrug-resistant fungal infections. PMID- 18385734 TI - A laser frequency comb that enables radial velocity measurements with a precision of 1 cm s(-1). AB - Searches for extrasolar planets using the periodic Doppler shift of stellar spectral lines have recently achieved a precision of 60 cm s(-1) (ref. 1), which is sufficient to find a 5-Earth-mass planet in a Mercury-like orbit around a Sun like star. To find a 1-Earth-mass planet in an Earth-like orbit, a precision of approximately 5 cm s(-1) is necessary. The combination of a laser frequency comb with a Fabry-Perot filtering cavity has been suggested as a promising approach to achieve such Doppler shift resolution via improved spectrograph wavelength calibration, with recent encouraging results. Here we report the fabrication of such a filtered laser comb with up to 40-GHz (approximately 1-A) line spacing, generated from a 1-GHz repetition-rate source, without compromising long-term stability, reproducibility or spectral resolution. This wide-line-spacing comb, or 'astro-comb', is well matched to the resolving power of high-resolution astrophysical spectrographs. The astro-comb should allow a precision as high as 1 cm s(-1) in astronomical radial velocity measurements. PMID- 18385732 TI - Imaging in the era of molecular oncology. AB - New technologies for imaging molecules, particularly optical technologies, are increasingly being used to understand the complexity, diversity and in vivo behaviour of cancers. 'Omic' approaches are providing comprehensive 'snapshots' of biological indicators, or biomarkers, of cancer, but imaging can take this information a step further, showing the activity of these markers in vivo and how their location changes over time. Advances in experimental and clinical imaging are likely to improve how cancer is understood at a systems level and, ultimately, should enable doctors not only to locate tumours but also to assess the activity of the biological processes within these tumours and to provide 'on the spot' treatment. PMID- 18385735 TI - Superinsulator and quantum synchronization. AB - Synchronized oscillators are ubiquitous in nature, and synchronization plays a key part in various classical and quantum phenomena. Several experiments have shown that in thin superconducting films, disorder enforces the droplet-like electronic texture--superconducting islands immersed into a normal matrix--and that tuning disorder drives the system from superconducting to insulating behaviour. In the vicinity of the transition, a distinct state forms: a Cooper pair insulator, with thermally activated conductivity. It results from synchronization of the phase of the superconducting order parameter at the islands across the whole system. Here we show that at a certain finite temperature, a Cooper--air insulator undergoes a transition to a superinsulating state with infinite resistance. We present experimental evidence of this transition in titanium nitride films and show that the superinsulating state is dual to the superconducting state: it is destroyed by a sufficiently strong critical magnetic field, and breaks down at some critical voltage that is analogous to the critical current in superconductors. PMID- 18385736 TI - Dust-climate couplings over the past 800,000 years from the EPICA Dome C ice core. AB - Dust can affect the radiative balance of the atmosphere by absorbing or reflecting incoming solar radiation; it can also be a source of micronutrients, such as iron, to the ocean. It has been suggested that production, transport and deposition of dust is influenced by climatic changes on glacial-interglacial timescales. Here we present a high-resolution record of aeolian dust from the EPICA Dome C ice core in East Antarctica, which provides an undisturbed climate sequence over the past eight climatic cycles. We find that there is a significant correlation between dust flux and temperature records during glacial periods that is absent during interglacial periods. Our data suggest that dust flux is increasingly correlated with Antarctic temperature as the climate becomes colder. We interpret this as progressive coupling of the climates of Antarctic and lower latitudes. Limited changes in glacial-interglacial atmospheric transport time suggest that the sources and lifetime of dust are the main factors controlling the high glacial dust input. We propose that the observed approximately 25-fold increase in glacial dust flux over all eight glacial periods can be attributed to a strengthening of South American dust sources, together with a longer lifetime for atmospheric dust particles in the upper troposphere resulting from a reduced hydrological cycle during the ice ages. PMID- 18385737 TI - Absolute plate motions and true polar wander in the absence of hotspot tracks. AB - The motion of continents relative to the Earth's spin axis may be due either to rotation of the entire Earth relative to its spin axis--true polar wander--or to the motion of individual plates. In order to distinguish between these over the past 320 Myr (since the formation of the Pangaea supercontinent), we present here computations of the global average of continental motion and rotation through time in a palaeomagnetic reference frame. Two components are identified: a steady northward motion and, during certain time intervals, clockwise and anticlockwise rotations, interpreted as evidence for true polar wander. We find approximately 18 degrees anticlockwise rotation about 250-220 Myr ago and the same amount of clockwise rotation about 195-145 Myr ago. In both cases the rotation axis is located at about 10-20 degrees W, 0 degrees N, near the site that became the North American-South American-African triple junction at the break-up of Pangaea. This was followed by approximately 10 degrees clockwise rotation about 145-135 Myr ago, followed again by the same amount of anticlockwise rotation about 110 100 Myr ago, with a rotation axis in both cases approximately 25-50 degrees E in the reconstructed area of North Africa and Arabia. These rotation axes mark the maxima of the degree-two non-hydrostatic geoid during those time intervals, and the fact that the overall net rotation since 320 Myr ago is nearly zero is an indication of long-term stability of the degree-two geoid and related mantle structure. We propose a new reference frame, based on palaeomagnetism, but corrected for the true polar wander identified in this study, appropriate for relating surface to deep mantle processes from 320 Myr ago until hotspot tracks can be used (about 130 Myr ago). PMID- 18385738 TI - A susceptibility locus for lung cancer maps to nicotinic acetylcholine receptor subunit genes on 15q25. AB - Lung cancer is the most common cause of cancer death worldwide, with over one million cases annually. To identify genetic factors that modify disease risk, we conducted a genome-wide association study by analysing 317,139 single-nucleotide polymorphisms in 1,989 lung cancer cases and 2,625 controls from six central European countries. We identified a locus in chromosome region 15q25 that was strongly associated with lung cancer (P = 9 x 10(-10)). This locus was replicated in five separate lung cancer studies comprising an additional 2,513 lung cancer cases and 4,752 controls (P = 5 x 10(-20) overall), and it was found to account for 14% (attributable risk) of lung cancer cases. Statistically similar risks were observed irrespective of smoking status or propensity to smoke tobacco. The association region contains several genes, including three that encode nicotinic acetylcholine receptor subunits (CHRNA5, CHRNA3 and CHRNB4). Such subunits are expressed in neurons and other tissues, in particular alveolar epithelial cells, pulmonary neuroendocrine cells and lung cancer cell lines, and they bind to N' nitrosonornicotine and potential lung carcinogens. A non-synonymous variant of CHRNA5 that induces an amino acid substitution (D398N) at a highly conserved site in the second intracellular loop of the protein is among the markers with the strongest disease associations. Our results provide compelling evidence of a locus at 15q25 predisposing to lung cancer, and reinforce interest in nicotinic acetylcholine receptors as potential disease candidates and chemopreventative targets. PMID- 18385742 TI - A single nucleotide polymorphism (A --> G) in intron 3 of IFNgamma gene is associated with asthma. AB - Interferon-gamma (IFNgamma) is located on chromosome 12, and a number of studies have detected very strong linkage signals around this gene and asthma. The aim of this study was to analyze the association of a (CA)n repeat in intron 1 and six single nucleotide polymorphisms (((rs2069705, T/C) (promoter)), ((rs1861494, A/G), (rs1861493, T/C), (rs2069718, C/T) (intron 3)), ((rs2069727, A/G) and (rs2069728, G/A) (3' untranslated region))) spanning the whole gene with asthma. We report here the association of rs1861494 A/G with atopic asthma in a case control cohort (n=189 and n=270 cases and controls, respectively) (P=0.0006), which was replicated (P=0.006) in a family study (n=137) as well. Allele G was found to be negatively associated (odds ratio=0.50, 95% confidence interval, P=0.0006). A five-locus haplotype also showed significant association with asthma in the case-control (P=0.002) and the family studies (P=0.0004). In our three locus sliding window haplotypic analysis, we found the (CA)n repeat, rs1861494 A/G and rs2069718 C/T to be of high priority (P=0.0003). Using electrophoretic mobility shift assay, we provide evidence that the alleles of rs1861494 A/G have differential affinity to bind to putative nuclear factor(s). In conclusion, we report for the first time association of rs1861494 A/G polymorphism with asthma, which may regulate the IFNgamma levels and, hence, modulate asthma pathogenesis. PMID- 18385740 TI - Cutaneous cancer stem cell maintenance is dependent on beta-catenin signalling. AB - Continuous turnover of epithelia is ensured by the extensive self-renewal capacity of tissue-specific stem cells. Similarly, epithelial tumour maintenance relies on cancer stem cells (CSCs), which co-opt stem cell properties. For most tumours, the cellular origin of these CSCs and regulatory pathways essential for sustaining stemness have not been identified. In murine skin, follicular morphogenesis is driven by bulge stem cells that specifically express CD34. Here we identify a population of cells in early epidermal tumours characterized by phenotypic and functional similarities to normal bulge skin stem cells. This population contains CSCs, which are the only cells with tumour initiation properties. Transplants derived from these CSCs preserve the hierarchical organization of the primary tumour. We describe beta-catenin signalling as being essential in sustaining the CSC phenotype. Ablation of the beta-catenin gene results in the loss of CSCs and complete tumour regression. In addition, we provide evidence for the involvement of increased beta-catenin signalling in malignant human squamous cell carcinomas. Because Wnt/beta-catenin signalling is not essential for normal epidermal homeostasis, such a mechanistic difference may thus be targeted to eliminate CSCs and consequently eradicate squamous cell carcinomas. PMID- 18385739 TI - A variant associated with nicotine dependence, lung cancer and peripheral arterial disease. AB - Smoking is a leading cause of preventable death, causing about 5 million premature deaths worldwide each year. Evidence for genetic influence on smoking behaviour and nicotine dependence (ND) has prompted a search for susceptibility genes. Furthermore, assessing the impact of sequence variants on smoking-related diseases is important to public health. Smoking is the major risk factor for lung cancer (LC) and is one of the main risk factors for peripheral arterial disease (PAD). Here we identify a common variant in the nicotinic acetylcholine receptor gene cluster on chromosome 15q24 with an effect on smoking quantity, ND and the risk of two smoking-related diseases in populations of European descent. The variant has an effect on the number of cigarettes smoked per day in our sample of smokers. The same variant was associated with ND in a previous genome-wide association study that used low-quantity smokers as controls, and with a similar approach we observe a highly significant association with ND. A comparison of cases of LC and PAD with population controls each showed that the variant confers risk of LC and PAD. The findings provide a case study of a gene-environment interaction, highlighting the role of nicotine addiction in the pathology of other serious diseases. PMID- 18385743 TI - Disturbed circadian blood pressure rhythm and C-reactive protein in essential hypertension. AB - We investigated the effect of the diverse definition criteria of the dipping and non-dipping status on the assessed differences in inflammatory activation between dippers and non-dippers with essential hypertension. 269 consecutive subjects (188 males, aged 50+/-7 years) with untreated stage I-II essential hypertension underwent ambulatory blood pressure (BP) monitoring and high-sensitivity C reactive protein (hs-CRP) level determination. The population was classified into dippers and non-dippers based on the three following different definitions: true non-dippers (TND): non-dippers (nocturnal fall of systolic and diastolic BP of <10% of the daytime values, n=95) and dippers (the remaining subjects, n=174); true dippers and true non-dippers (TD-TND): non-dippers (nocturnal fall of systolic and diastolic BP<10%, n=95) and dippers (nocturnal fall of systolic and diastolic BP> or =10%, n=75); systolic non-dippers (SND): non-dippers (nocturnal systolic BP fall of <10% of the daytime values, n=145) and dippers (the remaining subjects, n=124). Non-dippers compared to dippers in the TND, TD-TND and SND classification exhibited higher levels of log hs-CRP (by 0.11 mg l(-1), P=0.02; 0.13 mg l(-1), P=0.03 and 0.14 mg l(-1), P=0.02, respectively) and 24 h pulse pressure (PP) (by 4 mm Hg, P=0.006; by 5 mm Hg, P=0.003 and by 5 mm Hg, P<0.0001, respectively). Twenty-four hour PP and nocturnal systolic BP fall were independent predictors of log hs-CRP (P<0.05 for both) in multiple regression analysis. In conclusion, essential hypertensive non-dippers compared to dippers exhibit higher hs-CRP values, irrespective of the dipping status definition. Furthermore, ambulatory PP and nocturnal systolic BP fall interrelate and participate in the inflammatory processes that accompany non-dipping state. PMID- 18385745 TI - Obstructive sleep apnoea and metabolic syndrome: two sides of the same coin? PMID- 18385744 TI - Effects of eprosartan on mitochondrial membrane potential and H2O2 levels in leucocytes in hypertension. AB - We investigated whether circulating leucocytes from hypertensive patients exhibit more spontaneous, stimulated hydrogen peroxide (H2O2) production and greater mitochondrial membrane potential (Deltapsi) than those from normotensive individuals. We also investigated the effects of oral treatment with the angiotensin II (AT II) type 1 receptor blocker eprosartan (600 mg day(-1)) on these markers of oxidative stress. In 25 hypertensive patients and 28 healthy volunteers, spontaneous H2O2 formation was measured by flow cytometry after preincubation of buffy coat-leucocytes from fresh peripheral venous blood at 37 degrees C with 2',7' dichlorofluorescein. Stimulation of H2O2 formation by circulating leucocytes was elicited by the addition of tert-butylhydroperoxide (tBHP). Deltapsi was determined by flow cytometry after the addition of tetramethylrhodamine methyl ester (TMRM). Compared with healthy individuals, lymphocytes from hypertensive patients exhibited higher Deltapsi (12.28+/-3.20 vs 16.25+/-2.88 arbitrary fluorescence units (AFU), respectively; P<0.001) and greater spontaneous H2O2 production (4.75+/-5.15 vs 8.98+/-9.97 AFU, respectively; P<0.05). tBHP stimulation was associated with higher H2O2 levels in circulating leucocytes in patients with uncorrected hypertension than in normotensive individuals. H2O2 overproduction was corrected by eprosartan treatment. These results suggest that oxidative stress could be important in the pathogenesis of hypertension. Furthermore, measurement of leucocyte oxidant activities may be useful for the evaluation of oxidative stress, which may be reduced with the use of antihypertensive drugs. Our results demonstrate that treatment of hypertension with eprosartan normalizes blood pressure and corrects oxidative disturbances, suggesting that leucocytes could be a target for this drug. PMID- 18385746 TI - Hemoclip application for colonic diverticular hemorrhage. PMID- 18385749 TI - Interferon-alpha is able to maintain complete molecular remission induced by imatinib after its discontinuation. PMID- 18385751 TI - The peripheral helper T-cell repertoire in untreated indolent B-cell lymphomas: evidence for antigen-driven lymphomagenesis. PMID- 18385750 TI - A phase II study of 5-azacitidine for patients with primary and post-essential thrombocythemia/polycythemia vera myelofibrosis. AB - Myelofibrosis (MF; primary or post-essential thrombocythemia/polycythemia vera) is incurable clonal myeloproliferative disorder, with no effective treatment. Epigenetic changes play an important role in cancer pathogenesis through transcriptional silencing of critical tumor suppressor genes. We conducted a phase-II study to evaluate the activity of DNA methyltransferase inhibitor, 5 azacitidine, in patients with MF. Thirty-four patients (76% previously treated) received 5-azacitidine at 75 mg/m(2) subcutaneously daily for 7 days, every 4 weeks. Twelve (35%) patients had abnormal cytogenetics and 19 (70%) of 27 evaluable patients had JAK2(V617F) mutation. Responses occurred in 8 (24%) patients after a median of 5 months (range, 3-10). Partial response occurred in 1 (3%) patient (duration 22+ months) and clinical improvement in 7 (21%) patients (median duration 4 months; range, 2-8.5). Myelosuppression was the major adverse effect, with grade 3-4 neutropenia in 10 (29%) patients. Global DNA methylation assessed by the long interspersed nucleotide element (LINE) bisulfite/pyrosequencing assay decreased from 53% pretherapy to 44% on day 14 (P=0.0014) and returned to 50% at the end of the first 28-day cycle (P=0.016). 5 azacitidine is relatively well tolerated and results in induction of global hypomethylation in patients with MF, but results in limited clinical activity. PMID- 18385752 TI - The novel Akt inhibitor, perifosine, induces caspase-dependent apoptosis and downregulates P-glycoprotein expression in multidrug-resistant human T-acute leukemia cells by a JNK-dependent mechanism. AB - A significant impediment to the success of cancer chemotherapy is the occurrence of multidrug resistance, which, in many cases, is attributable to overexpression of membrane transport proteins, such as the 170-kDa P-glycoprotein (P-gp). Also, upregulation of the phosphatidylinositol 3-kinase (PI3K)/Akt-signaling pathway is known to play an important role in drug resistance, and has been implicated in the aggressiveness of a number of different cancers, including T-acute lymphoblastic leukemia (T-ALL). We have investigated the therapeutic potential of the novel Akt inhibitor, perifosine (a synthetic alkylphospholipid), on human T ALL CEM cells (CEM-R), characterized by both overexpression of P-gp and constitutive upregulation of the PI3K/Akt network. Perifosine treatment induced death by apoptosis in CEM-R cells. Apoptosis was characterized by caspase activation, Bid cleavage and cytochrome c release from mitochondria. The proapoptotic effect of perifosine was in part dependent on the Fas/FasL interactions and c-Jun NH(2)-terminal kinase (JNK) activation, as well as on the integrity of lipid rafts. Perifosine downregulated the expression of P-gp mRNA and protein and this effect required JNK activity. Our findings indicate that perifosine is a promising therapeutic agent for treatment of T-ALL cases characterized by both upregulation of the PI3K/Akt survival pathway and overexpression of P-gp. PMID- 18385753 TI - Global increase in DNA methylation in patients with myelodysplastic syndrome. PMID- 18385755 TI - Life expectancy and prognostic factors in the classic BCR/ABL-negative myeloproliferative disorders. AB - Among the 'classic' BCR/ABL-negative chronic myeloproliferative disorders, primary myelofibrosis (PMF) is associated with a substantial life-expectancy reduction. In this disease, initial haemoglobin level is the most important prognostic factor, whereas age, constitutional symptoms, low or high leukocyte counts, blood blast cells and cytogenetic abnormalities are also of value. Several prognostic systems have been proposed to identify subgroups of patients with a different risk, which is especially important in younger individuals, who may benefit from therapies with curative potential. Essential thrombocythaemia (ET) affects the patients' quality of life more than the survival, due to the high occurrence of thrombosis, whereas polycythaemia vera (PV) has a substantial morbidity derived from thrombosis but also a certain reduction in life expectancy. Therefore, in the latter disorders, prognostic studies have focused primarily on prediction of the thrombosis, with age and a previous history of thrombosis being the main prognostic factors of such complication. The importance of higher leukocyte counts in thrombosis development has been recently pointed out in ET and PV, where a role for mutated JAK2 allele burden has also been noted. With regard to PMF, the possible association of the mutation with shorter survival and higher acute transformation rate is currently being evaluated. PMID- 18385754 TI - Effective killing of Gleevec-resistant CML cells with T315I mutation by a natural compound PEITC through redox-mediated mechanism. AB - Mutation of Bcr-Abl is an important mechanism by which chronic myelogenous leukemia (CML) cells become resistant to Gleevec. The T315I mutation is clinically significant since CML cells harboring this mutation are insensitive to Gleevec and other Bcr-Abl-targeted drugs. Identification of new agents capable of effectively killing CML cells with T315I mutation would have important therapeutic implications in Gleevec-resistant CML. Here, we showed that beta phenylethyl isothiocyanate (PEITC), a natural compound found in vegetables, is effective in killing CML cells expressing T315I BCR-ABL. Treatment of leukemia cell lines harboring wild-type or mutant Bcr-Abl with 10 microM PEITC resulted in an elevated ROS stress and a redox-mediated degradation of the BCR-ABL protein, leading to massive death of the leukemia cells. Antioxidant NAC attenuated the PEITC-induced oxidative stress in CML cells and prevented the degradation of BCR ABL, caspase-3 activation and cell death. We further showed that the ROS-induced degradation of BCR-ABL was mediated partially by caspase-3 and the proteasome pathway. The ability of PEITC to effectively kill T315I-positive CML cells was further confirmed using primary leukemia cells isolated from CML patients. Our results suggest that PEITC is a promising compound capable of killing Gleevec resistant CML cells through a ROS-mediated mechanism and warrants further investigations. PMID- 18385756 TI - Two cell lines of t(8;21) acute myeloid leukemia with activating KIT exon 17 mutation: models for the 'second hit' hypothesis. PMID- 18385757 TI - Activation of keratinocyte protein kinase C zeta in psoriasis plaques. AB - PKCzeta (protein kinase C-zeta), a member of protein kinase C family, plays an important role in cell proliferation, differentiation, and apoptosis. It acts as a downstream molecule for TNF-alpha (tumor necrosis factor) signal transduction and also regulates the expression of CD1d, an HLA-class I-like molecule. The interaction of CD1d with natural killer T (NKT) cells has been shown to be important in their Th1 cytokine production in psoriasis. In this study, we examined PKCzeta in psoriasis in order to define its role in the pathogenesis of the disease. We found that T-cell receptor (TCR) V alpha24+ V beta11+ NKT cells and CD1d molecules within psoriatic skin were increased. Moreover, there was an associated increase in PKCzeta mRNA and protein expression with membrane translocation in psoriasis lesions compared to uninvolved skin. Furthermore, cultured keratinocytes exhibited increased PKCzeta activity and membrane translocation upon stimulation by TNF-alpha, a cytokine known to play an important role in the pathogenesis of psoriasis. These results implied that PKCzeta is an important transduction molecule downstream of TNF-alpha signaling and is associated with increased expression of CD1d that may enhance CD1d-NKT cell interactions in psoriasis lesions. This makes PKCzeta a tempting target for possible pharmacological intervention in modifying the downstream effects of TNF alpha in psoriasis. PMID- 18385758 TI - Potential of fibroblast cell therapy for recessive dystrophic epidermolysis bullosa. AB - Recessive dystrophic epidermolysis bullosa (RDEB) is a severe inherited skin blistering disorder caused by mutations in the COL7A1 gene that lead to reduced type-VII collagen and defective anchoring fibrils at the dermal-epidermal junction (DEJ). Presently there are no effective treatments for this disorder. Recent mouse studies have shown that intradermal injections of normal human fibroblasts can generate new human type-VII collagen and anchoring fibrils at the DEJ. To assess potential clinical benefits in humans, we gave single intradermal injections of allogeneic fibroblasts to five subjects with RDEB. We noted increased type-VII collagen at the DEJ at 2 weeks and at 3 months following injection and increased anchoring fibrils, although none of these had normal morphology. No adverse effects, clinical or immunopathologic, were noted. We believe the major effect of allogeneic fibroblasts is to increase the recipients' own COL7A1 mRNA levels with greater deposition of mutant type-VII collagen at the DEJ and formation of additional rudimentary anchoring fibrils. Nevertheless, this mutant protein may be partially functional and capable of increasing adhesion at the DEJ. This is the first study demonstrating that intradermal injections of allogeneic fibroblasts have therapeutic potential in human subjects with RDEB. PMID- 18385759 TI - Th2 cytokines act on S100/A11 to downregulate keratinocyte differentiation. AB - Atopic dermatitis (AD) is an inflammatory skin disease associated with frequent skin infection and impaired skin barrier function. Recent studies indicate that increased Th2 cytokine expression contributes to reduction in antimicrobial peptides and reduced filaggrin (FLG) expression, however, the mechanisms leading to this effect is unknown. Using proteomics, we found the S100 calcium-binding protein A11 (S100/A11) to be significantly downregulated in the presence of IL-4 and IL-13. Culturing keratinocytes with increased calcium concentrations significantly induced S100/A11 expression. This corresponded with an increase in human beta-defensin (HBD)-3 and FLG expression. Interference of S100/A11 expression, by siRNA, inhibited induction of HBD-3 and FLG. Furthermore p21, a cyclin-dependent kinase inhibitor downstream of S100/A11, was required for calcium-mediated induction of HBD-3 and FLG. Importantly, transduction of p21 recombinant protein into keratinocytes prevented IL-4/IL-13-mediated inhibition of FLG and HBD-3 expression. S100/A11 and p21 gene expression was also found to be significantly lower in acute and chronic AD skin. This study demonstrates an important role for S100/A11 and p21 in regulating skin barrier integrity and the innate immune response. PMID- 18385760 TI - Penile intraepithelial neoplasia is frequent in HIV-positive men with anal dysplasia. AB - Anogenital human papillomavirus (HPV)-infection is common in HIV-infected men who have sex with men (HIV+MSM). These patients have a strongly increased risk of HPV induced anal cancer and its precursor lesion, anal intraepithelial neoplasia (AIN), and a moderately increased risk for penile cancer. Only limited data exist on penile intraepithelial neoplasia (PIN) in HIV+MSM. We determined the prevalence and evaluated the virologic characteristics of PIN and AIN in 263 HIV+MSM. In case of histologically confirmed PIN (and AIN), HPV-typing, HPV-DNA load determination, and immunohistochemical staining for p16(INK4a) were performed. PIN was detected in 11 (4.2%) and AIN in 156 (59.3%) patients. Ten PIN patients also had AIN within the observation period. Four clinical types of PINs could be distinguished. High-risk-alpha-HPV-DNA was found in 10 PIN lesions, with HPV16 being the most frequent type. Infections with multiple HPV-types were common. All high-grade lesions had high-risk-HPV-DNA-loads > or = 1 HPV-copy/beta globin-gene-copy. Cutaneous beta-HPVs were found in PIN and AIN, but beta-HPV-DNA loads were very low, irrespective of the histological grade. p16(INK4a) Expression was detectable in all PIN lesions and correlated both with the histological grade and with high-risk HPV-DNA loads. In view of the PIN prevalence found in our study, all HIV+MSM should be screened for PIN in addition to AIN screening. PMID- 18385761 TI - Humoral autoimmune responses to the squamous cell carcinoma antigen protein family in psoriasis. AB - Substantial evidence indicates that psoriasis is a T-lymphocyte-mediated autoimmune disease. However, longstanding data also indicate IgG and complement deposition in upper epidermis of psoriasis plaques. This led us to propose that autoantigen-autoantibody interactions in the skin may also be of pathogenic importance. Here, we have confirmed the presence of IgG in upper lesional epidermis and used high-resolution two-dimensional immunoblotting of extracts from this tissue, and laser desorption mass spectrometry of tryptic peptides, to define a series of epidermal proteins that bind IgG from psoriatic serum. The most prominent of these autoantigens are homologues of the serpin, squamous cell carcinoma antigen (SCCA), the other autoantigens identified including arginase 1, enolase 1, and keratin 10. Blood levels of IgG autoantibodies that bind to SCCA proteins were significantly higher in psoriasis than healthy controls (P=0.005), but were not detectable in sera from patients with active atopic dermatitis. To our knowledge, SCCA proteins have not previously been described as autoantigenic in animals or humans and form complexes with IgG that are associated with complement deposition. These findings expose potentially pathogenic humoral immunologic events and thus possible therapeutic targets in psoriasis. PMID- 18385762 TI - K6PC-5, a direct activator of sphingosine kinase 1, promotes epidermal differentiation through intracellular Ca2+ signaling. AB - Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, regulates multiple cellular responses such as Ca(2+) signaling, growth, survival, and differentiation. Because sphingosine kinase (SphK) is the enzyme directly responsible for production of S1P, many factors have been identified that regulate its activity and subsequent S1P levels. Here we synthesized a previously unidentified SphK activator, K6PC-5, and have studied its effects on intracellular Ca(2+) signaling in HaCaT cells and epidermal differentiation in murine skin. K6PC-5, a hydrophobic compound chemically named N-(1,3 dihydroxyisopropyl)-2-hexyl-3-oxo-decanamide, activated SphK (obtained from C57BL/6 murine blood and F9-12 cell lysates) in a dose-dependent manner. K6PC-5 induced both intracellular Ca(2+) concentration ([Ca(2+)](i)) oscillations in HaCaT cells and Ca(2+) mobilization in hairless mouse epidermis. Both dimethylsphingosine (DMS) and dihydroxysphingosine (DHS), SphK inhibitors, and transfection of SphK1-siRNA blocked K6PC-5-induced increases in [Ca(2+)](i). The K6PC-5-induced [Ca(2+)](i) oscillations were dependent on thapsigargin-sensitive Ca(2+) stores and Ca(2+) entry, but independent of the classical phospholipase C mediated pathway. In addition, K6PC-5 enhanced the expression of involucrin and filaggrin, specific differentiation-associated marker proteins in HaCaT cells, whereas transfection of SphK1-siRNA blocked the increase of involucrin. Topical K6PC-5 also enhanced the expression of involucrin, loricrin, filaggrin, and keratin 5 in intact murine epidermis. Finally, topical K6PC-5 inhibited epidermal hyperplasia by exerting antiproliferative effects on keratinocytes in murine epidermis. These results suggest that K6PC-5 acts to regulate both differentiation and proliferation of keratinocytes via [Ca(2+)](i) responses through S1P production. Thus, regulation of S1P levels may represent a novel approach for treatment of skin disorders characterized by abnormal differentiation and proliferation, such as atopic dermatitis and psoriasis. PMID- 18385763 TI - EDA2R is associated with androgenetic alopecia. AB - Androgenetic alopecia (AGA) is a common heritable polygenic disorder whose genetics is not fully understood, even though it seems to be X-linked. We carried out an epidemiological survey for AGA on 9,000 people from 8 isolated villages of a secluded region of Sardinia (Ogliastra), and identified a large cohort of affected individuals. We genotyped 200 cases and 200 controls (mean kinship 0.001) with the 500k chip array and conducted case-control association analysis on the X chromosome. We identified Xq11-q12 as strongly associated with AGA. In particular, we found that rs1352015 located 8 kb from the EDA2R gene showed the best result (P=7.77e(-7)). This region also contains the AR gene, hence we tested both genes in 492 cases and 492 controls. We found that the non-synonymous SNP rs1385699 on EDA2R gave the best result (P=3.9e(-19)) whereas rs6152 on the AR gene is less significant (P=4.17e(-12)). Further statistical analysis carried out by conditioning each gene to the presence of the other showed that the association with EDA2R is independent while the association with AR seems to be the result of linkage disequilibrium. These results give insight into the pathways involved in AGA etiology. PMID- 18385764 TI - Restoration of the expression of transports associated with antigen processing in human malignant melanoma increases tumor-specific immunity. AB - The transporter associated with antigen processing (TAP) is essential for peptide delivery from the cytosol into the lumen of the endoplasmic reticulum (ER), where these peptides are loaded on HLA I molecules. Our previous study found that expressions of TAP were reduced in human malignant melanoma (MM) lesions and associated with histopathologic characteristics. In this study, we further investigate expressions of TAP and HLA class I antigen in three human MM cell lines. pEGFP-TAP1/TAP2/TAP1+TAP2 were used to restore the expressions of TAP in the antigen presentation pathway-deficient MM cell line A375. TAP1- and TAP1+TAP2 transfected A375 increased TAP1, TAP2, and HLA class I antigen expression and antigen presentation. TAP1- and TAP1+TAP2-transfected A375 exhibited a dramatic increase in Melan-A-specific cytotoxic T lymphocytes (CTLs) compared with TAP2 transfected A375 or empty vector. These CTLs were capable of killing TAP1- and TAP1+TAP2-transfected A375. TAP1+TAP2-transfected A375 generated the highest frequency of Melan-A-specific IL-12 and interferon (IFN)-gamma-producing CD8+ T cells compared with TAP1, TAP2, and empty vector. Therefore, TAP expression restores both antigen presentation and immunogenicity in A375 melanoma cells and concomitantly increases IL-12 and IFN-gamma production in tumor antigen-specific CTLs; TAP should be considered as a part of the immunotherapies for MM. PMID- 18385765 TI - Immunity to adeno-associated virus vectors in animals and humans: a continued challenge. AB - Recombinant vectors based on adeno-associated virus (AAV) have been shown to stably express many genes in vivo without mounting immune responses to vectors or transgenes. Thus, AAV vectors have rapidly become the reagents of choice for therapeutic gene transfer. Yet one of the first translations of AAV gene therapy into humans unexpectedly resulted in only short-term expression of the therapeutic gene accompanied by transient but significant toxicity. Immune responses to the vector capsid were held accountable for these results, confirming that a detailed understanding of the interaction of AAV vectors with the immune system is of great importance for the safety and success of gene therapy applications. Most humans display naturally acquired immunity to AAV; circumventing neutralizing antibodies and memory T-cell responses is challenging, but not impossible. This review will evaluate the strategies that have been proposed to overcome such responses and summarize recent findings about the mechanisms and circumstances that lead to the activation of innate and adaptive immune responses to AAV vector components. PMID- 18385766 TI - Ultrasound with microbubbles enhances gene expression of plasmid DNA in the liver via intraportal delivery. AB - Current ultrasound (US)-mediated gene delivery methods are inefficient due, in part, to a lack of US optimization. We systematically explored the use of microbubbles (MBs), US parameters and plasmid delivery routes to improve gene transfer into the mouse liver. Co-presentation of plasmid DNA (pDNA), 10% Optison MBs and pulsed 1-MHz US at a peak negative pressure of 4.3 MPa significantly increased luciferase gene expression with pDNA delivered by intrahepatic injection to the left liver lobe. Intraportal injection delivered pDNA and MBs to the whole liver; with insonation, all lobes expressed the transgene, thus increasing total gene expression. Gene expression was also dependent on acoustic pressure over the range of 0-4.3 MPa, with a peak effect at 3 MPa. An average of 85-fold enhancement in gene delivery was achieved. No enhancement was observed below 0.25 MPa. Increasing pulse length while decreasing pulse repetition frequency and exposure time to maintain a constant total energy during exposure did not further improve transfection efficiency, nor did extend the US exposure pre- or postinjection of pDNA. The results indicate that coupled with MBs, US can more efficiently and dose-dependently enhance gene expression from pDNA delivered via portal vein injection by an acoustic mechanism of inertial cavitation. PMID- 18385767 TI - Tissue-specific restriction of cyclophilin A-independent HIV-1- and SIV-derived lentiviral vectors. AB - The host factor alpha isoform of the tripartite motif 5 (TRIM5alpha) restricts human immunodeficiency virus type 1 (HIV-1) infection in certain non-human primate species. Restriction of HIV-1 is enhanced by binding of the viral capsid to cyclophilin A (CypA) in target cells, although CypA is not absolutely required for restriction in rhesus macaque cells. Simian immunodeficiency virus (SIV) is not restricted by rhesus macaque TRIM5alpha and its capsid does not bind to CypA. Here, the effect of lentiviral CypA dependence on restriction in different tissues was examined by engineering an HIV-1 capsid quadruple mutant (V(86)P/H(87)Q/I(91)V/M(96)I) lentiviral vector (HIV(quad)) that is CypA independent. Whereas HIV-1 was restricted in rhesus macaque and owl monkey epithelial cells, infection with the HIV(quad) vector was efficient at high viral concentrations. In contrast, HIV(quad) was largely restricted in primary rhesus macaque CD34(+) cells. Human epithelial and primary CD34(+) cells were permissive for HIV-1, HIV(quad) and SIV, whereas transduction of human T cells by HIV(quad) or SIV was impaired. The restrictive human cells did not express increased levels of TRIM5alpha, and restriction was not relieved by abolishing CypA, consistent with HIV(quad) and SIV being CypA-independent. Pseudotyping of lentiviral vectors with the gibbon ape leukemia virus envelope altered their sensitivity to perturbations of the virus-CypA interaction compared to pseudotyping with vesicular stomatitis virus glycoproteins, suggesting that the viral entry pathway modulates restriction. Together, these studies reveal that an HIV-1 capsid quadruple mutant can partially overcome lentiviral restriction in non-human primate epithelial cells, but not in hematopoietic cells. Similarly, human cells vary in their permissiveness for CypA-independent lentiviruses, and suggest the presence of tissue-specific factor(s) that can inhibit lentiviral transduction independently of viral interaction with TRIM5alpha and CypA. PMID- 18385768 TI - Cells as vehicles for therapeutic genes to treat liver diseases. AB - Gene therapy involves the transfer of genetic sequences to tissues to obtain a curative effect. Effective gene transfer can be achieved by introducing the therapeutic gene into virus-like particles that facilitate the penetration of the transgene into the cells. However, direct injection of viral vectors may activate innate immunity leading to toxic effects. On the other hand, viral vectors frequently induce neutralizing antibodies, which limit the efficacy of repeated vector administration. Moreover, targeting of the transgene to the desired tissue is a goal that not always can be attained with current vectors. The use of cells as vehicles for therapeutic genes may offer solutions for these issues. Ex vivo transduction of specific cells with vectors encoding therapeutic genes followed by injection of the engineered cells to the patient will reduce the inherent toxicity of the vector while preventing the development of neutralizing antibodies. At the same time, this therapeutic approach can take advantage of the homing properties of the transduced cells to target transgene expression to the sites of interest. Thus, it has been shown that administration of dendritic cells engineered ex vivo with vectors encoding selected antigenic determinants or immunostimulatory molecules is an efficient means to elicit protective immune responses. Similarly, since endothelial progenitor cells (EPC) move to inflammed, ischemic or neoplastic tissues, the injection of EPC transduced ex vivo with appropriate therapeutic genes is an effective method to direct transgene expression to the lesions to be treated. Promising data in animal models of disease point to a future clinical application of this therapeutic strategy. PMID- 18385769 TI - Gene-modified bone marrow cell therapy for prostate cancer. AB - There is a critical need to develop new and effective cancer therapies that target bone, the primary metastatic site for prostate cancer and other malignancies. Among the various therapeutic approaches being considered for this application, gene-modified cell-based therapies may have specific advantages. Gene-modified cell therapy uses gene transfer and cell-based technologies in a complementary fashion to chaperone appropriate gene expression cassettes to active sites of tumor growth. In this paper, we briefly review potential cell vehicles for this approach and discuss relevant gene therapy strategies for prostate cancer. We further discuss selected studies that led to the conceptual development and preclinical testing of IL-12 gene-modified bone marrow cell therapy for prostate cancer. Finally, we discuss future directions in the development of gene-modified cell therapy for metastatic prostate cancer, including the need to identify and test novel therapeutic genes such as GLIPR1. PMID- 18385770 TI - Characterizing mixed microbial population dynamics using time-series analysis. AB - Due to a general shortage of temporal population data, dynamic structures in microbial communities remain largely unexplored. Knowledge of community dynamics is, however, essential for understanding the mechanisms by which microbes interact. Here, we have used a computational approach for quantification of bacteria in multispecies populations, generating data for time-series modeling. Moreover, we have used online FR-IR spectroscopy to monitor the main metabolic processes. The approach enabled us to provide a functional description of the parameters governing the population dynamics in a three-species model bacterial community, demonstrating density-dependent regulation, interspecies competition and even a case of cooperation between two species. Since the field of microbial ecology has yet to embrace many of the concepts and methods developed for the study of ecology of higher plants and animals, the realization that microbial systems can be analyzed within the same conceptual framework as other ecosystems is of fundamental importance. PMID- 18385771 TI - Applying stable isotope probing of phospholipid fatty acids and rRNA in a Chinese rice field to study activity and composition of the methanotrophic bacterial communities in situ. AB - Methanotrophs in the rhizosphere play an important role in global climate change since they attenuate methane emission from rice field ecosystems into the atmosphere. Most of the CH(4) is emitted via transport through the plant gas vascular system. We used this transport for stable isotope probing (SIP) of the methanotrophs in the rhizosphere under field conditions and pulse-labelled rice plants in a Chinese rice field with CH(4) (99% (13)C) for 7 days. The rate of (13)CH(4) loss rate during (13)C application was comparable to the CH(4) oxidation rate measured by the difluoromethane inhibition technique. The methanotrophic communities on the roots and in the rhizospheric soil were analyzed by terminal-restriction fragment length polymorphism (T-RFLP), cloning and sequencing of the particulate methane monooxygenase (pmoA) gene. Populations of type I methanotrophs were larger than those of type II. Both methane oxidation rates and composition of methanotrophic communities suggested that there was little difference between urea-fertilized and unfertilized fields. SIP of phospholipid fatty acids (PLFA-SIP) and rRNA (RNA-SIP) were used to analyze the metabolically active methanotrophic community in rhizospheric soil. PLFA of type I compared with type II methanotrophs was labelled more strongly with (13)C, reaching a maximum of 6.8 atom-%. T-RFLP analysis and cloning/sequencing of 16S rRNA genes showed that methanotrophs, especially of type I, were slightly enriched in the 'heavy' fractions. Our results indicate that CH(4) oxidation in the rice rhizosphere under in situ conditions is mainly due to type I methanotrophs. PMID- 18385772 TI - Global-scale processes with a nanoscale drive: the role of marine viruses. PMID- 18385774 TI - Impact of measurement precision and noise on superresolution image reconstruction. AB - The performance of uniform and nonuniform detector arrays for application to the PANOPTES (processing arrays of Nyquist-limited observations to produce a thin electro-optic sensor) flat camera design is analyzed for measurement noise environments including quantization noise and Gaussian and Poisson processes. Image data acquired from a commercial camera with 8 bit and 14 bit output options are analyzed, and estimated noise levels are computed. Noise variances estimated from the measurement values are used in the optimal linear estimators for superresolution image reconstruction. PMID- 18385775 TI - Time-resolved polarimetry over water waves: relating glints and surface statistics. AB - The phenomenon of sunglint, well known in satellite remote sensing, lacks a fundamental characterization under controlled laboratory conditions. Exploiting an apparatus specifically assembled for the purpose, we examine the signal collected by a photopolarimeter, pointed at a wavy water surface with measurable statistics and illuminated by a laser source. We also analyze the wave slope distributions, retrieved with an imaging system, and correlate them with the time series of glints. More particularly, we investigate the link between the occurrence of glints and that of the slopes from which they originate. In this context, the results obtained by applying the Hilbert-Huang transform technique to the slope time series are compared with those obtained through a traditional Fourier transform. This novel study first identifies the individual atomic glints as Fresnel reflection originating from a single wave facet. It then discusses the periodic character of a sequence of glints generated by a gravity wave state, as opposed to the erratic behavior of glints typical of capillary wave states. In mixed gravity-capillary conditions, it is shown that the glint properties are governed mainly by the capillary regime. PMID- 18385776 TI - Ritonavir-boosted protease inhibitor monotherapy for the treatment of HIV-1 infection. AB - Guidelines for the use of antiretrovirals for HIV-1 infection recommend combining at least three agents. Toxicities, cost, and the complexity of such regimens warrant the search for other options. Boosted protease inhibitor monotherapy is one of the appealing options being investigated. Herein we review uncontrolled and controlled clinical trials evaluating boosted protease inhibitor monotherapy in several clinical settings: maintenance therapy, induction-maintenance strategies, and first-line treatment. Boosted lopinavir monotherapy has been largely investigated in maintenance and induction-maintenance strategies, showing its ability to maintain viral suppression in the majority of participants. The major concern is the higher proportion of patients experiencing transient episodes of low-level viremia (HIV-RNA 50-500 copies/ml) when compared to classical triple regimens. No protease inhibitor-associated resistance mutation was detected in patients who failed on boosted lopinavir monotherapy. Three uncontrolled maintenance strategy studies with boosted atazanavir monotherapy showed conflicting results. Thus, the reassuring results obtained with lopinavir might not be extended to the whole protease inhibitor class, warranting further studies with new generation protease inhibitors such as darunavir. Finally, one controlled trial comparing first-line boosted lopinavir monotherapy to a standard triple combination showed that the latter outperformed the boosted protease inhibitor monotherapy in this clinical setting. In summary, a boosted protease inhibitor single-agent strategy can maintain continuous plasma HIV-RNA suppression in a large proportion of patients already suppressed on a standard triple combination. The more frequent occurrence of low-level viremia, however, does not allow the widespread use of such a strategy outside of clinical studies at this time. PMID- 18385777 TI - Pathogenesis of liver damage in HCV-HIV patients. AB - The coinfection of HIV and HCV has become a pathology with several distinctive characteristics. Pathogenesis of liver damage in patients with HIV and HCV coinfection is complex and multifactorial. It derives from a balance of factors which interact among themselves in a dynamic way. The reasons for the accelerated course of HIV/HCV coinfection are mainly related to two principal causes: the persistence of HCV, which depends upon alterations of cell-mediated immunity, and the activation of the immune system towards secretion of proinflammatory and profibrotic cytokines. This review will first focus on the characteristics of both these immune-mediated mechanisms, and then their implication on fibrogenesis as well as on other pathogenetic mechanisms, such as interactions between viruses and the deficit of protective mechanisms. A better knowledge of adaptive immune mechanisms, cytokine alteration, interference with host defense mechanisms, and the "cross-talk" among the viruses will improve the understanding of the pathogenetic mechanism and provide the opportunity to cure this disease. PMID- 18385778 TI - Multicentric Castleman's disease in HIV infection: a systematic review of the literature. AB - The objective of this study is to systematically review the epidemiology and the clinical and virologic aspects of multicentric Castleman's disease in HIV positive patients and to evaluate treatment strategies and outcome, especially in relation to HAART administration. The authors have conducted a systematic review of the English literature for all cases of newly diagnosed multicentric Castleman's disease in HIV-positive patients. The 25 studies which met the selection criteria included 84 HIV-positive patients with multicentric Castleman's disease (20 pre-HAART and 64 post-HAART era). Of them, the majority (90%) were men with 33 months median time from detection of HIV-positivity to multicentric Castleman's disease diagnosis in the HAART era. Fever and lymphadenopathy were the most common presenting symptoms and cytopenias, hypoalbuminemia, polyclonal hypergammaglobulinemia and raised C-reactive protein the most frequently revealed laboratory findings. Kaposi's sarcoma was present in 72% of the patients and respiratory system involvement in 34%. Although the majority of cases reported were positive for human herpesvirus-8, none of the reviewed patients was found to suffer from polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS) syndrome. Of the 48 patients on HAART, 64% were already on HAART at multicentric Castleman's disease diagnosis, having a better immunologic profile and a lower incidence of Kaposi's sarcoma than the 35% of patients who initiated HAART after multicentric Castleman's disease diagnosis. Nevertheless, the two groups did not have significantly different mortality rates (30 vs. 38%). At multicentric Castleman's disease diagnosis, a wide range of CD4 counts was recorded, suggesting that disease presentation could occur at any CD4 count. With regard to treatment, the study confirmed the high rates of response with rituximab (anti-CD20 monoclonal). Monochemotherapy seems to give short-lived responses, which require maintenance to be sustained. Polychemotherapy with CHOP has given long-term remission in a subset of patients. Other regimens used in the treatment of HIV-related multicentric Castleman's disease were antiviral agents, immunomodulatory agents, and thalidomide. The fatality rate among HIV-related multicentric Castleman's disease cases reviewed was 44%, significantly lower than that of HIV-negative individuals (65%), while median survival of the latter was 29 months longer than that of HIV-infected individuals. The fatality rate among pre-HAART patients was 75 vs. 29% among HAART patients. Infection, multiorgan failure, Kaposi's sarcoma, non-Hodgkin lymphoma and progressive multicentric Castleman's disease were the most often reported causes of death. In conclusion, multicentric Castleman's disease is a lymphoproliferative disorder with an increasing prevalence in HIV infected individuals. Even though life expectancy in multicentric Castleman's disease seems to have significantly improved in the HAART era, it remains a disease with a poor prognosis and an increased incidence of non-Hodgkin lymphoma in the HIV-context. PMID- 18385779 TI - Mucosal immune dysfunction in AIDS pathogenesis. AB - The mucosal immune system plays a central role in both the transmission of HIV infection and the pathogenesis of AIDS. Most HIV infections are acquired through mucosal transmission, and quantitative and qualitative defects of mucosal immunity are consistently present in all stages of pathogenic HIV and SIV infections. A series of recent studies has emphasized the role of a rapid, dramatic, and largely irreversible depletion of mucosa-associated lymphoid tissue based memory CD4(+)CCR5(+) T-cells as a key determinant of disease progression in HIV-infected individuals and SIV-infected macaques. It has also been proposed that, in order to be effective, an AIDS vaccine should prevent the early depletion of these mucosal CD4(+) T-cells. However, the observation of depletion of mucosal CD4(+) T-cells during the primary phase of nonpathogenic SIV infection of natural SIV hosts, such as sooty mangabeys and African green monkeys, suggests that additional pathogenic factors are involved in the AIDS-associated mucosal immune dysfunction. These factors may include: (i) selective depletion of specific CD4(+) T-cell subsets; (ii) dysfunction of other (non-CD4(+)) immune cells; and (iii) generalized immune activation. Importantly, the mucosal immune dysfunction observed during pathogenic HIV and SIV infection is associated with translocation of microbial products (i.e. lipopolysaccharide) from the intestinal lumen to the systemic circulation where they may be responsible, at least in part, for the chronic immune activation that follows pathogenic HIV and SIV infections. The role of mucosal immunity in AIDS pathogenesis emphasizes the importance of understanding whether and to what extent the HIV-associated depletion of mucosal CD4(+) T-cells is reversible after prolonged suppression of virus replication with antiretroviral therapy. Further studies of mucosal immunity during primate lentiviral infections will be needed to better understand, and ultimately prevent and treat, the mechanisms underlying the AIDS associated mucosal immune dysfunction. PMID- 18385780 TI - Nosocomial HIV infection: epidemiology and prevention--a global perspective. AB - Because, globally, HIV is transmitted mainly by sexual practices and intravenous drug use and because of a long asymptomatic period, healthcare-associated HIV transmission receives little attention even though an estimated 5.4% of global HIV infections result from contaminated injections alone. It is an important personal issue for healthcare workers, especially those who work with unsafe equipment or have insufficient training. They may acquire HIV occupationally or find themselves before courts, facing severe penalties for causing HIV infections. Prevention of blood-borne nosocomial infections such as HIV differs from traditional infection control measures such as hand washing and isolation and requires a multidisciplinary approach. Since there has not been a review of healthcare-associated HIV contrasting circumstances in poor and rich regions of the world, the aim of this article is to review and compare the epidemiology of HIV in healthcare facilities in such settings, followed by a consideration of general approaches to prevention, specific countermeasures, and a synthesis of approaches used in infection control, injury prevention, and occupational safety. These actions concentrated on identifying research on specific modes of healthcare-associated HIV transmission and on methods of prevention. Searches included studies in English and Russian cited in PubMed and citations in Google Scholar in any language. MeSH keywords such as nosocomial, hospital-acquired, iatrogenic, healthcare associated, occupationally acquired infection and HIV were used together with mode of transmission, such as "HIV and hemodialysis". References of relevant articles were also reviewed. The evidence indicates that while occasional incidents of healthcare-related HIV infection in high-income countries continue to be reported, the situation in many low-income countries is alarming, with transmission ranging from frequent to endemic. Viral transmission in health facilities occurs by unexpected and unusual as well as more frequent modes. HIV can be transmitted to patients and to donors of blood products by specific vehicles and vectors during blood transfusion, plasma donation, and artificial insemination, by improperly sterilized sharps, by medical equipment during activities such as dialysis and organ transplantation, and by healthcare workers infected by occupational exposure to hazards such as blood-contaminated sharps. Personal, equipment, and environmental factors predispose to acquisition of nosocomial HIV and all are pertinent for prevention. For infection and injury control, poverty is often an underlying determinant. While sophisticated new tests offer improved HIV detection, increasingly higher marginal costs limit their feasibility in many settings. Modest investment in safer equipment and appropriate integrated training in infection control, injury prevention, and occupational safety should provide greater benefit. PMID- 18385781 TI - Absence of optineurin (OPTN) gene mutations in Taiwanese patients with juvenile onset open-angle glaucoma. AB - PURPOSE: To investigate sequence variants in the optineurin (OPTN) gene in patients with juvenile-onset open-angle glaucoma (JOAG) in Taiwan. METHODS: We analyzed the sequence variants of OPTN in 51 unrelated Taiwanese probands with JOAG and in 51 control group subjects who did not have JOAG. Genomic DNA was extracted from the individuals and subjected to polymerase chain reaction (PCR) to amplify all 16 exons and flanking introns of OPTN. The amplified products were then screened for base variants by autosequence. Data from the two study groups were then compared using Fisher's exact test and Armitage's trend test. RESULTS: Fifteen variants of OPTN were found in the 51 JOAG patients and 51 unrelated normal controls. Two were missense variants (M98K and K322E), one was a synonymous codon change (T34T), and 12 were changes in the noncoding sequences. Seven of the variants have been reported and eight were novel. All of the sequence changes were found in patients with JOAG and in the normal controls except for variant c.-233+25C>G, which was found only in the control group. Allelic frequencies of these sequence changes did not differ significantly between patients and controls (p>0.05) except for the variant c.-233+25C>G (p<0.001). Genotype frequencies of c.-233+25C>G was shown to be significant between the two groups using Fisher's two-tailed exact test (p<0.001) and Armitage's trend test (p=6.815e(-06)). CONCLUSIONS: Our data indicate that none of the mutations in OPTN are associated with JOAG. The variant M98K is not a risk factor and the variant c.-233+25C>G may be protective against glaucoma in Taiwanese. PMID- 18385782 TI - Atypical asymmetric lattice corneal dystrophy associated with a novel homozygous mutation (Val624Met) in the TGFBI gene. AB - PURPOSE: To evaluate the TGFBI gene and the encoded transforming growth factor beta-induced protein (TGFBIp) in a 47-year-old African-American patient with an unusual atypical asymmetric lattice corneal dystrophy (LCD). METHODS: The eyes of the proband and his brother were examined by slit-lamp biomicroscopy and their clinical records were reviewed. All 17 exons of TGFBI were evaluated in genomic DNA extracted from blood or buccal epithelial cell samples from the proband and his family members. The corneal tissue of the proband was examined histopathologically, and TGFBIp was analyzed in half of an excised corneal button. RESULTS: The proband (who had an unusual atypical asymmetric LCD) and his brother (who had mild bilateral deep stromal opacities) were found to have homozygous Val624Met mutations in TFGBI. The proband's daughter who was heterozygous for the Val624Met mutation had no reported ophthalmic abnormalities. The corneal tissue from the proband contained TGFBIp with the Val624Met mutation. Patients with LCD have different clinical phenotypes based on their genotype. Molecular genetic analyses are becoming increasingly important in making precise diagnoses and prognostic predictions about inherited corneal disorders. CONCLUSIONS: A novel Val624Met homozygous mutation in TGFBI was associated with atypical LCD in two family members. Symptomatic corneal disease was absent at the age of 24 years in the offspring of the proband who was heterozygous for this mutation. This is an apparent example of a TGFBI mutation that becomes evident when it is homozygous. The finding of Val624Met mutated TGFBIp in a approximately 65 kDa protein band in a reduced sodium dodecyl sulfate (SDS) gel suggests that the accumulated protein was intact TGFBIp and not a fragment of TGFBIp. PMID- 18385783 TI - Comparative analysis of the tear protein profile in mycotic keratitis patients. AB - PURPOSE: Mycotic keratitis is a major cause of corneal blindness in India. A proper understanding of the pathogenesis may help in refining the existing treatment. The purpose of this study is to examine the total tear protein profile of fungal keratitis patients, which may have a bearing on pathogenesis and disease progression. METHODS: Tear samples were collected from culture positive fungal keratitis patients. Tears from the uninfected fellow eye and from other healthy individuals served as controls. Two-dimensional electrophoresis (2DE) was used for the separation of fractionated tear proteins, and selected protein spots, which showed differential expressions, were identified using matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Wherever needed, tag sequencing of peptide fragments using post source decay (PSD) was done to confirm the identification. RESULTS: The glutaredoxin-related protein was expressed only in the tears of fungal keratitis patients. Six other normal tear proteins were present in both samples but with varied expression levels. Prolactin inducible protein and serum albumin precursor were upregulated in the infected samples. Cystatin S precursor, cystatin SN precursor, cystatin, and human tear lipocalin were downregulated in the infected samples. CONCLUSIONS: Tears can be used as a clinical source to study the proteomic responses in patients with fungal keratitis. The glutaredoxin-related protein is known to be produced by Aspergillus during oxidative stress conditions, and the presence of this protein in the tears of patients with mycotic keratitis indicates that this pathogen undergoes stress-related gene expression during infection. PMID- 18385784 TI - Contributions of MYOC and CYP1B1 mutations to JOAG. AB - PURPOSE: To investigate the role of MYOC and CYP1B1 in Iranian juvenile open angle glaucoma (JOAG) patients. METHODS: Twenty-three JOAG probands, their available affected and unaffected family members, and 100 ethnically matched control individuals without history of ocular disease were recruited. Clinical examinations of the probands included slit lamp biomicroscopy, intraocular pressure (IOP) measurement, gonioscopic evaluation, fundus examination, and perimetry measurement. Familial cases were classified according to the mode of inheritance. Exons of MYOC and CYP1B1 were sequenced, and novel variations assessed in the control individuals. Potential disease-associated variations were tested for segregation with disease status in available family members. RESULTS: The mode of inheritance of the disease in the families of four probands (17.4%) appeared to be autosomal dominant and in at least eight (34.8%) to be autosomal recessive. Four patients carried MYOC mutations, and an equal number carried CYP1B1 mutations. The MYOC mutations were heterozygous; two of them (p.C8X and p.L334P) are novel, and one codes for the shortest truncated protein so far reported. Autosomal recessive inheritance was consistent with inheritance observed in families of patients carrying CYP1B1 mutations. All these patients carried homozygous mutations. CONCLUSIONS: MYOC and CYP1B1 contributed equally to the disease status of the Iranian JOAG patients studied. The contribution of the two genes appeared to be independent in that no patient carried mutations in both genes. The fraction of Iranian patients carrying MYOC mutations was comparable to previously reported populations. PMID- 18385785 TI - The role of mitochondrial haplogroups in glaucoma: a study in an Arab population. AB - PURPOSE: Glaucoma prevalence can vary geographically and ethnically, which suggests that a genetic element could play a significant role. Studies investigating the role of various mitochondrial haplogroups in the pathogenesis of glaucoma are scarce. METHODS: We compared the prevalence of different mitochondrial haplogroups in 107 glaucoma patients (49 primary open-angle glaucoma, POAG; 29 primary angle-closure glaucoma, PACG; and 29 pseudoexfoliation glaucoma, PEG) and 552 maternally unrelated normal controls. All patients and controls were Saudi Arabs. RESULTS: There was no statistically significant difference between patients and controls for all mitochondrial haplogroups tested except for PACG patients with mitochondrial haplogroup preHV1 (odds ratio=4.9; 95% CI 2.3 - 10.5; p=0.00002). CONCLUSIONS: Patients with preHV1 mitochondrial haplogroups are at higher risk of developing PACG. However, our study group is relatively small and further studies with more patients in other populations are needed to confirm this interesting finding. PMID- 18385786 TI - Distribution of adenosine receptors in human sclera fibroblasts. AB - PURPOSE: Systemic treatment with adenosine receptor antagonists has been reported to affect the biochemistry and ultrastructure of rabbit sclera. This study was conducted to determine whether adenosine receptors (ADORs) are present in human scleral fibroblasts (HSF). METHODS: Primary HSF were cultured in vitro and identified with anti-vimentin, anti-keratin, anti-desmin, and anti-S-100 antibodies. Confocal fluorescence microscopy was used to study the distribution of ADORs in the HSF cell lines and in the frozen human scleral sections. ADOR protein expression in HSF and human sclera was confirmed by western blot analysis of cell lysates. RESULTS: ADORs were expressed in both HSF and human sclera. This was confirmed by western blot. ADORA1 expression was concentrated in the nucleus. ADORA2A was concentrated mainly in one side of the cytoplasm, and ADORA2B was found both in the nucleus and the cytoplasm. ADORA3 was expressed weakly in the cytoplasm. CONCLUSIONS: All four subtypes of ADOR were found in HSF and may play a role in scleral remodeling. PMID- 18385787 TI - Autosomal dominant congenital cataract in a Libyan Jewish family: cosegregation with a reciprocal chromosomal translocation [t(3;5)(p22.3; p15.1)]. AB - PURPOSE: To describe a Jewish family of Libyan ancestry in which autosomal dominant congenital cataract segregates with an apparently balanced reciprocal chromosomal translocation. METHODS: Detailed family history and clinical data were recorded. Cytogenetic studies were performed on 13 family members. RESULTS: Embryonal cataracts cosegregated through three generations with a balanced chromosomal translocation [t(3;5)(p22.3; p15.1)] while the unbalanced translocation product, 46,XY,-5,+der(5)t(3:5)(p22:p15.1), had multiple congenital anomalies without cataracts. CONCLUSIONS: These observations suggest that an altered function of a gene at one of the translocation breakpoints on chromosome 3p22.3 or 5p15.1 is causally related to cataract development. PMID- 18385788 TI - Evaluation of LOXL1 gene polymorphisms in exfoliation syndrome and exfoliation glaucoma. AB - PURPOSE: To evaluate genetic susceptibility of lysyl oxidase-like 1 (LOXL1) gene polymorphisms to exfoliation syndrome (XFS) and exfoliation glaucoma (XFG) in a case-control cohort of American and European patients. METHODS: DNA from a total of 620 individuals including 287 exfoliation patients and 333 healthy control subjects were extracted by standard methods. Three single nucleotide polymorphisms (SNPs) of rs1048661 (R141L), rs3825942 (G153D), and rs2165241 were genotyped in these individuals by SNaPshot Assay. The seven coding exons of the LOXL1 gene and their immediate flanking regions were directly sequenced in 95 affected patients. Data management and case-control association studies were performed with SNP-STAT and PLINK programs. The obtained DNA sequences were evaluated with the STADEN package. RESULTS: The 287 unrelated exfoliation cases comprised of 171 American patients (mostly of European background) and 116 patients from 12 European countries. This phenotype was further divided into patients with exfoliation only and no glaucoma (XFO; n=95), exfoliation with glaucoma (XFG; n=133), and exfoliation unclassified (XFU; n=59). Genotypic data were analyzed separately for XFO, XFG, XFU, and XFS (all exfoliations; n=287) and for Americans and Europeans. The observed genotypic frequencies for each exfoliation phenotype or population were tabulated separately and tested for deviation from the Hardy-Weinberg equilibrium (HWE) using a standard Chi(2) test. There were no HWE deviations and no significant genotypic differences between these subcategories for the three studied SNPs. For the combined exfoliation cohort, homozygote genotypes of G/G (rs1048661), G/G (rs3825942), and T/T (rs2165241) were significantly overrepresented. Likewise, case-control allelic association for rs1048661 (p=7.74x10(-9)), rs3825942 (p=3.10x10(-17)), and rs2165241 (p=4.85x10(-24)) were highly significant. The corresponding two-locus haplotype frequencies of GG for rs1048661-rs3825942 (p=1.47x10(-27)), GT for rs1048661-rs2165241 (p=1.29x10(-24)), and GT for rs3825942-rs2165241 (p=2.02x10( 24)) were highly associated with exfoliation phenotypes. The combined effect of these three SNPs revealed that the GGT haplotype is overrepresented by 66% in exfoliation cases, and this deviation from controls is highly significant (p=1.93x10(-24)). This haplotype constituted a major risk factor for development of exfoliation in both XFS and XFG. By contrast, the GAC haplotype was significantly underrepresented (p=4.99x10(-18)) in exfoliation cases by 83% and may potentially have a protective effect for this condition with an estimated attributable risk percent reduction of 457%. The only other haplotype that was significantly different between cases and controls was TGC (p=5.82x10(-9)). No observation was made for the GAT haplotype. The combined three haplotypes of GGT, GAC, and TGC were associated with 91% of the exfoliation syndrome cases in the studied populations. Seven coding exons of LOXL1 were also sequenced in 95 affected cases. In addition to the three above-mentioned SNPs, 12 other variations were also observed in these patients (G240G, D292D, A320A, V385V, rs2304719, IVS3+23C>T, IVS3-155G>A, IVS3-101G>A, IVS4+49G>A, rs2304721, IVS5 121C>T, and rs2304722). None were considered a disease-causing mutation. CONCLUSIONS: We confirmed a strong association with LOXL1 variants in our patients. For the LOXL1 gene, individual alleles of rs1048661 (G), rs3825942 (G), and rs2165241 (T) are highly associated with XFS and XFG in American and European populations. The GGT haplotype constitutes a major risk haplotype for exfoliation, and GAC may have a protective role. DNA sequencing of 95 affected patients did not show any mutations in this gene. The LOXL1 SNPs are located in the 15q24.1 band and within a genetic locus (GLC1N) that is associated with primary open-angle glaucoma (POAG). However, the LOXL1 genetic predisposition is only limited to exfoliation with or without glaucoma and does not include the POAG phenotype. PMID- 18385790 TI - HLA class I and II gene polymorphisms in Stevens-Johnson syndrome with ocular complications in Japanese. AB - PURPOSE: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are acute-onset mucocutaneous diseases induced by infectious agents and/or inciting drugs. Although the pathobiological mechanisms underlying the onset of SJS/TEN have not been fully established, the extreme rarity of cutaneous and ocular surface reactions to drug therapies led us to suspect individual susceptibility. Our previous study of polymorphisms in the HLA-class I genes of 40 Japanese SJS/TEN patients with ocular surface complications showed that in the Japanese, HLA-A*0206 was strongly associated with SJS/TEN. In this study, we investigated the association between HLA class II antigens in addition to HLA class I antigens and SJS/TEN. METHODS: We studied the histocompatibility antigen genes, HLA-A, B, C, DRB1, and DQB1, of 71 Japanese SJS/TEN patients with ocular complications. We also genotyped 113 healthy volunteers for HLA-A, B, C, DRB1, and DQB1. We performed polymerase chain reaction amplification followed by hybridization with sequence-specific oligonucleotide probes (PCR-SSO) using commercial bead-based typing kits. RESULTS: HLA-A*0206 was strongly associated with SJS/TEN. HLA-A*1101 was inversely associated. HLA-B*5901 exhibited a high odds ratio for SJS/TEN with ocular complications. However, when we corrected the p-value for the number of alleles detected (n=29), the results ceased to be significant. There was no association between HLA-C and SJS/TEN. There was also no significant association between HLA-DRB1 and SJS/TEN. HLA-DQB1*0502 was negatively and weakly associated with SJS/TEN although correction of the p-value for the number of alleles detected rendered the result not significant. CONCLUSIONS: Because our findings are completely different from data reported on Caucasian patients, they suggest strong ethnic differences in the HLA-SJS associations. PMID- 18385789 TI - The Cdk5 inhibitor olomoucine promotes corneal debridement wound closure in vivo. AB - PURPOSE: To investigate the effect of the Cdk5 inhibitor olomoucine on corneal debridement wound healing in vivo. METHODS: Corneal debridement wounds of 1.5 mm were made on the ocular surface of CD-1 mice. A 20 microl drop of 15 microM olomoucine in 1% DMSO was applied to the wound area immediately after wounding and again after 6 h. Control mice received identical applications of 1% DMSO. Mice were euthanized after 18 h, two weeks, and three weeks for evaluation of wound healing and restratification. Corneas were stained with Richardson's dye, photographed, and processed for histology and immunofluorescence as whole mounts or paraffin sections. The remaining wound area at 18 h was measured by image analysis. Scratch wounded cultures of human corneal-limbal epithelial cells (HCLE) were used to examine the effect of olomoucine on matrix metalloproteinase (MMP) expression in vitro. MMP-2 and MMP-9 were detected by immunofluorescence and immunoblotting. RESULTS: Olomoucine treatment significantly enhanced corneal wound closure without increasing inflammation or infiltration of polymorphonuclear leukocytes 18 h after wounding (p<0.05). The increased localization of MMP-9 within epithelial cells at the wound edge was further enhanced by olomoucine while the expression of MMP-2 was reduced. Olomoucine treatment of scratch wounded HCLE cells produced similar changes in MMP-9 and MMP 2 expression. The examination of treated corneas two and three weeks after wounding showed normal epithelial restratification with no evidence of inflammation or stromal disorganization. CONCLUSIONS: Topical application of olomoucine in 1% DMSO significantly enhances closure of small epithelial debridement wounds without increasing inflammation or impairing reepithelialization. PMID- 18385791 TI - Inhibition of choroidal neovascularization by homoisoflavanone, a new angiogenesis inhibitor. AB - PURPOSE: Age-related macular degeneration (AMD) is the leading cause of blindness in elderly. The detailed mechanism of choroidal neovascularization (CNV) leads to severe vision loss in patients with AMD. This study was undertaken to evaluate the inhibitory effect of homoisoflavanone on CNV. METHODS: Antiangiogenic activity of homoisoflavanone was evaluated by in vitro tube formation assay of human umbilical vein endothelial cells (HUVECs) and cell migration assay of HUVECs., Homoisoflavanone or PBS was injected intravitreously into a mouse model of laser-photocoagulation-induced CNV. Fluorescein angiography and vessel counting in cross sections were employed to examine CNV lesions. The toxicity of homoisoflavanone was evaluated through 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay (MTT) assay in HUVECs as well as histological examination and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) staining in the retina. RESULTS: Homoisoflavanone effectively inhibited in vitro tube formation and cell migration of HUVECs. Interestingly, homoisoflavanone significantly reduced CNV and its leakage in a mouse model of laser-photocoagulation-induced CNV. In addition, homoisoflavanone showed no effect on cell viability of HUVECs and no retinal toxicity in a concentration range of 1-10 microM. CONCLUSIONS: Our data suggest that homoisoflavanone is a potent inhibitor of CNV and may be applied in the treatment of other vasoproliferative retinopathies and tumor. PMID- 18385792 TI - Promiscuous recombination of LoxP alleles during gametogenesis in cornea Cre driver mice. AB - PURPOSE: To examine whether promiscuous Cre/LoxP recombination happens during gametogenesis in double transgenic mice carrying LoxP modified alleles and Cre transgene driven by tissue-specific promoter outside the gonads of adult mice. METHODS: Cre driver mice were crossbred with reporter mouse lines (e.g., ZEG and Rosa26R) to obtain Cre/ZEG and Cre/Rosa26R double transgenic mice. The frequency of promiscuous LoxP/Cre recombination was determined by the expression of second reporter genes in the offspring of double transgenic mice. RESULTS: The frequency of promiscuous LoxP/Cre recombination varied in different lines of Cre driver mice and in the sex of the same driver mice with higher penetrance in male than in female double transgenic mice. Polymerase chain reaction (PCR) and recombination analysis demonstrate that the recombination of floxed allele occurs during the transition from spermatogonia (diploid) to primary spermatocyte (tetraploid) in the testis. Thereby, target-floxed allele(s) may be ubiquitously ablated in experimental animals intended for tissue-specific gene deletion. CONCLUSIONS: Gametogenesis-associated recombination should always be examined in tissue-specific gene ablation studies. PMID- 18385793 TI - Mie light scattering calculations for an Indian age-related nuclear cataract with a high density of multilamellar bodies. AB - PURPOSE: Multilamellar bodies (MLBs) are lipid-coated spheres (1-4 microm in diameter) found with greater frequency in the nuclear region of human age-related cataracts compared with human transparent lenses. Mie light scattering calculations have demonstrated that MLBs are potential sources of forward light scattering in human age-related nuclear cataracts due to their shape, size, frequency, and cytoplasmic contents, which often differ in refractive index from their surroundings. Previous studies have used data from several non-serial tissue sections viewed by light microscopy to extrapolate a volume and have assumed that MLBs are random in distribution. Currently, confocal microscopy is being used to examine actual tissue volumes from age-related nuclear cataracts and transparent lenses collected in India to confirm MLB shape, size, frequency, and randomness. These data allow Mie scattering calculations to be done with directly observed MLBs in intact tissue. METHODS: Whole Indian donor lenses and Indian lens nuclei after extracapsular cataract extraction were immersion-fixed in 10% formalin for 24 h and in 4% paraformaldehyde for 24 h before sectioning with a Vibratome. The 160 microm thick sections were stained for 24 h in the lipid dye DiI (1,1'-dilinoleyl-3,3,3',3' tetramethylindocarbocyanine, 4 chlorobenzenesulfonate), washed, stabilized in Permount under coverslips and examined with a Zeiss LSM 510 confocal microscope. Individual volumes of tissue (each typically 500,000 microm(3)) were examined using a plan-apochromat 63X oil (NA=1.4) lens. Other lenses were prepared for electron microscopy and histological examination using previously described procedures. RESULTS: Analysis of tissue volumes within Indian age-related nuclear cataracts and transparent lenses has confirmed that most MLBs are 1-4 microm in diameter and typically spherical with some occurring as doublets or in clusters. Most Indian cataracts and transparent lenses are similar to samples obtained in the United States. One cataract contained as many as 400,000 MLBs per mm(3) -100 times more than in cataracts collected in the United States. Pairwise distribution analysis has revealed that MLBs even in this exceptional case are found with a distribution that appears to be random. Mie calculations indicate that more than 90% of the incident light could be scattered by the high density of MLBs. CONCLUSIONS: An important finding was that one advanced Indian cataract contained many more MLBs than cataracts examined from India and previously from the United States. This indicates that specific conditions or susceptibilities may exist that promote the formation of excessive MLBs. Based on the extremely high frequency, as well as their spherical shape, large size, and apparent random distribution, the MLBs are predicted according to Mie light scattering calculations to cause high amounts of forward scattering sufficient to produce nuclear opacity. PMID- 18385794 TI - Identification of novel mutations and sequence variants in the SOX2 and CHX10 genes in patients with anophthalmia/microphthalmia. AB - PURPOSE: Mutations in the SOX2 and CHX10 genes have been reported in patients with anophthalmia and/or microphthalmia. In this study, we evaluated 34 anophthalmic/microphthalmic patient DNA samples (two sets of siblings included) for mutations and sequence variants in SOX2 and CHX10. METHODS: Conformational sensitive gel electrophoresis (CSGE) was used for the initial SOX2 and CHX10 screening of 34 affected individuals (two sets of siblings), five unaffected family members, and 80 healthy controls. Patient samples containing heteroduplexes were selected for sequence analysis. Base pair changes in SOX2 and CHX10 were confirmed by sequencing bidirectionally in patient samples. RESULTS: Two novel heterozygous mutations and two sequence variants (one known) in SOX2 were identified in this cohort. Mutation c.310 G>T (p. Glu104X), found in one patient, was in the region encoding the high mobility group (HMG) DNA-binding domain and resulted in a change from glutamic acid to a stop codon. The second mutation, noted in two affected siblings, was a single nucleotide deletion c.549delC (p. Pro184ArgfsX19) in the region encoding the activation domain, resulting in a frameshift and premature termination of the coding sequence. The shortened protein products may result in the loss of function. In addition, a novel nucleotide substitution c.*557G>A was identified in the 3'-untranslated region in one patient. The relationship between the nucleotide change and the protein function is indeterminate. A known single nucleotide polymorphism (c. *469 C>A, SNP rs11915160) was also detected in 2 of the 34 patients. Screening of CHX10 identified two synonymous sequence variants, c.471 C>T (p.Ser157Ser, rs35435463) and c.579 G>A (p. Gln193Gln, novel SNP), and one non-synonymous sequence variant, c.871 G>A (p. Asp291Asn, novel SNP). The non-synonymous polymorphism was also present in healthy controls, suggesting non-causality. CONCLUSIONS: These results support the role of SOX2 in ocular development. Loss of SOX2 function results in severe eye malformation. CHX10 was not implicated with microphthalmia/anophthalmia in our patient cohort. PMID- 18385795 TI - Erythrocyte aldose reductase activity and sorbitol levels in diabetic retinopathy. AB - PURPOSE: Activation of polyol pathway due to increased aldose reductase (ALR2) activity has been implicated in the development of diabetic complications including diabetic retinopathy (DR), a leading cause of blindness. However, the relationship between hyperglycemia-induced activation of polyol pathway in retina and DR is still uncertain. We investigated the relationship between ALR2 levels and human DR by measuring ALR2 activity and its product, sorbitol, in erythrocytes. METHODS: We enrolled 362 type 2 diabetic subjects (T2D) with and without DR and 66 normal subjects in this clinical case-control study. Clinical evaluation of DR in T2D patients was done by fundus examination. ALR2 activity and sorbitol levels along with glucose and glycosylated hemoglobin (HbA1C) levels in erythrocytes were determined. RESULTS: T2D patients with DR showed significantly higher specific activity of ALR2 as compared to T2D patients without DR. Elevated levels of sorbitol in T2D patients with DR, as compared to T2D patients without DR, corroborated the increased ALR2 activity in erythrocytes of DR patients. However, the increased ALR2 activity was not significantly associated with diabetes duration, age, and HbA1C in both the DR group and total T2D subjects. CONCLUSIONS: Levels of ALR2 activity as well as sorbitol in erythrocytes may have value as a quantitative trait to be included among other markers to establish a risk profile for development of DR. PMID- 18385796 TI - Overexpression of E2F5/p130, but not E2F5 alone, can inhibit E2F-induced cell cycle entry in transgenic mice. AB - PURPOSE: The retinoblastoma (Rb) gene family member p130 binds preferentially to the E2F5 transcription factor and forms a repressive E2F5/p130 complex that inhibits cell cycle progression and tumor growth. It is unclear whether E2F5, either alone or in combination with p130, can interfere with the transcriptional activity of other E2F family members, such as E2F1 and E2F3a, in vivo. In this study, we used transgenic mice to test whether overexpression of E2F5 with/without p130 would be sufficient to inhibit E2F1 or E2F3a induced cell cycle reentry. METHODS: Transgenic mice were generated by microinjection of constructs containing different E2F cDNAs (E2F1, E2F3a, and E2F5) or the p130 cDNA linked to the mouse alphaA-crystallin promoter. The E2F5 single and E2F5/p130 double transgenic mice were cross-mated with E2F1 or E2F3a transgenic mice. The resulting double or triple transgenic mouse embryos were characterized by histology, in situ hybridization, immunohistochemistry, and BrdU incorporation assays. RESULTS: Overexpression of E2F5 alone was not sufficient to inhibit E2F1 or E2F3a induced cell cycle reentry in lens fiber cells. Transgenic mice coexpressing E2F5 and p130 in lens fiber cells did not show lens defects. However, coexpression of E2F5/p130 with E2F1 or E2F3a in lens fiber cells decreased the number of BrdU positive fiber cells induced by the E2F1 or E2F3a alone. Therefore, overexpression of E2F5/p130, but not E2F5 alone, can inhibit activator E2F-mediated cell proliferation in vivo, confirming that p130 plays a critical role in the repressive activity of E2F5/p130 complex. CONCLUSIONS: Overexpression of E2F5/p130 in post-mitotic lens fiber cells does not affect their normal differentiation program, but can inhibit inappropriate cell cycle reentry induced by the activator E2Fs. Since E2F5 alone cannot interfere with these E2F activities, we conclude that p130 is a key player in the inhibitory process. PMID- 18385797 TI - Characterization of intraocular immunopathology following intracameral inoculation with alloantigen. AB - PURPOSE: Anterior chamber-associated immune deviation (ACAID) is a form of peripheral tolerance achieved via intracameral antigen inoculation. It is well known that ACAID effectively down-regulates potentially destructive immunities such as delayed-type hypersensitivity (DTH) at extraorbital sites. However, what has not been specifically addressed is whether local intraocular tissues are negatively affected from intracamerally placed antigen. Thus, the current study was undertaken to detect and characterize potential pathological effects on intraocular tissues following intracameral inoculation with alloantigen. METHODS: ACAID induced in C57BL/6 hosts via intracameral inoculation with allogeneic (BALB/c) splenocytes was confirmed by the absence of DTH reactivity in the periphery. Injuries to the anterior segment and neuroretina following intracameral inoculation were evaluated pathologically via histological evaluation, molecularly via upregulation of glial fibrillary acidic protein (GFAP), and functionally via assessment of photoreceptor degeneration and electroretinogram (ERG) out to 24 days. In all experiments, intracamerally inoculated mice were compared to sham-operated, and controlled lens-punctured mice--a procedure that elicits intracameral inflammation for positive identification of immunopathological changes. RESULTS: Inflammation of anterior segment tissues persisted out to eight days, despite evidence that significant clearance of allogeneic cells took place within 6 h. In the neuroretina, a transient loss in ERG B-wave amplitudes was detected, but photoreceptor degeneration and GFAP upregulation were not. CONCLUSIONS: Intracameral inoculation with alloantigen leads to anterior segment inflammation and ERG dysfunction; however, this was markedly reduced and transient when compared to strong anterior segment inflammation induced by a more serious lens-puncture wound. PMID- 18385799 TI - Multiplex bead analysis of vitreous and serum concentrations of inflammatory and proangiogenic factors in diabetic patients. AB - PURPOSE: To investigate the role of inflammatory and angiogenic factors in the pathogenesis of diabetic retinopathy, we determined, in diabetic patients and controls, vitreous and serum concentrations of interferon-induced protein (IP) 10, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, regulated upon activation, normal T-expressed and secreted (RANTES), and vascular endothelial growth factor (VEGF). METHODS: We recruited 36 probands with type 2 diabetes mellitus (15 noninsulin-dependent and 21 insulin-dependent) and 69 normal controls. Using Cytometric Bead Array Technology, we measured vitreous and serum concentrations of IP-10, MCP-1, MIP 1alpha, MIP-1beta, RANTES, and VEGF. RESULTS: In diabetic patients the mean vitreous levels of IP-10, MCP-1 and VEGF were significantly higher compared normal controls. [IP-10 (pg/mL) 254.84 +/-311.67 versus 78.90 +/- 67.94 (p<0.001); MCP-1 (pg/mL) 1127.14 +/- 738.91 versus 700.80 +/-419.21 (p=0.002); VEGF (pg/mL) 954.98 +/- 2315.09 versus 37.90 +/- 28.51(p<0.001)]. Vitreous levels of VEGF correlated with vitreous levels of both IP-10 and MCP-1 (p<0.05). MIP 1beta, RANTES, and VEGF mean serum levels were significantly raised in diabetic probands while IP-10, MCP-1, and MIP-1alpha serum levels showed no significant elevation compared to controls [IP-10 (pg/mL) 346.20 +/- 287.36 versus 328.74 +/ 352.35 (p=0.88); MCP-1(pg/mL) 133.10 +/- 89.10 versus 141.47 +/- 222.15 (p=0.50); MIP-1beta (pg/mL) 184.40 +/- 100.20 versus 139.56 +/- 151.38 (p=0.003); RANTES (pg/mL) 51336.23 +/- 19940.31 versus 33629.2 +/- 33301.0 (p=0.002); VEGF (pg/mL) 304.88 +/- 257.52 versus 154.45 +/- 114.78 (p<0.001)]. CONCLUSIONS: Our results suggest that in diabetics, there is an upregulation of IP-10, MCP-1, and VEGF in the vitreous and an upregulation of MIP-1beta, RANTES, and VEGF in the serum. These findings support the concept of an angiogenic and inflammatory element in the development of diabetic retinopathy. PMID- 18385798 TI - Beta1-6 branching of cell surface glycoproteins may contribute to uveal melanoma progression by up-regulating cell motility. AB - PURPOSE: This study investigated the influence of integrin expression as well as the oligosaccharide structure of surface N-glycoproteins on cell behavior of two primary uveal (92-1 and Mel202) and two primary cutaneous (FM55P and IGR-39) melanoma cell lines. METHODS: Cell adhesion to fibronectin and cell migration on fibronectin (wound healing) were selected as the studied cell behavior parameters. The percentage of cells positive for expression of selected integrins was estimated by flow cytometric analysis. The influence of beta1-6 branched complex-type N-oligosaccharides on wound healing on fibronectin was investigated. Cell surface beta1-6 branched N-oligosaccharides were measured by their specific binding to PHA-L followed by flow cytometry, and the fibronectin receptors bearing beta1-6 GlcNAc branched N-linked glycans were identified. In addition, the transcript of GnT-V (the enzyme that catalyzes the addition of N acetylglucosamine to the core mannose of di- and tri-antennary N-glycans through a beta1-6 linkage) was analyzed by semiquantitative RT-PCR. RESULTS: Unlike the two examined cutaneous melanoma cell lines, neither of the uveal melanoma cells adhered to fibronectin. The adhesion efficiency of IGR-39 cells was twice that of FM55P cells. In contrast, uveal melanoma cells repaired scratch wounds on fibronectin-coated surfaces twice as fast as cutaneous melanoma cells did. The expression of alpha(3)beta(1), alpha(4)beta(1), alpha(5)beta(1), and alpha(v)beta(3) integrins, acting as fibronectin receptors, differed between the tested cell lines, and no distinct pattern distinguished uveal melanoma from cutaneous melanoma except for high expression of alpha(4)beta(1) integrin on both FM55P and IGR-39 cells. The results also demonstrated that the high levels of alpha(3)beta(1), alpha(4)beta(1), and alpha(5)beta(1) integrin expression on IGR 39 cells promoted their strong attachment to fibronectin-coated surfaces. In addition, 92-1, Mel202, and FM55P cells showed no or low adhesion to fibronectin, perhaps the result of low expression of fibronectin receptors excluding high expression of alpha(4)beta(1) integrin in FM55P cells. Cell migration was significantly decreased in three out of four PHA-L-treated cell lines, suggesting that beta1-6 branched complex type N-oligosaccharides are critical for 92-1, Mel202, and FM55P cell motility. Semiquantitative RT-PCR analysis showed that the tested cells did not differ in mRNA levels of beta1-6 -N acetylglucosaminyltransferase V. However, FACS analysis showed that 92-1, Mel202 and IGR-39 cells expressed significantly higher amounts of beta1-6 branched N oligosaccharides on the cell surface than FM55P cells did. All examined alpha(3), alpha(5), alpha(v), and beta(1) integrin subunits were shown to bear beta1-6 branched N-linked glycans. CONCLUSIONS: The role of integrins and their N glycosylation in the regulation of uveal melanoma growth and progression is largely unknown. These results reveal that cell surface complex-type N-glycans with GlcNAc beta1-6 branches are important factors determining the migration of primary uveal melanoma cells on fibronectin. PMID- 18385800 TI - Human retinal Muller cells synthesize collagens of the vitreous and vitreoretinal interface in vitro. AB - PURPOSE: To investigate the capacity of cultured Muller cells to synthesize collagens, since previous studies indicated that Muller cells could be involved in collagen remodeling at the vitreoretinal border in adult human eyes. METHODS: Spontaneously immortalized cultured human Muller cells were analyzed for the presence of mRNA of types I-VII, IX, XI, and XVII collagen by RT-PCR. Furthermore, Muller cells were immunocytochemically stained for light microscopic (LM) evaluation of these collagens and their main characteristics. Finally, cell extracts and culture medium were evaluated by western blot (WB) analysis using anticollagen antibodies. RESULTS: Cultured Muller cells contained mRNA for types I-VII, IX, and XI collagen, but not for type XVII collagen. LM and WB confirmed the intracellular expression of all the above-mentioned collagens with the exception of type XVII. Collagen secretion into the medium was established for types I-VII, IX, and XI collagen. CONCLUSIONS: Cultured Muller cells can synthesize internal limiting lamina and vitreous collagens. Possible collagen production by Muller cells could explain and expand on previous in vivo morphological findings in the embryonic and postnatal period and in pathologic conditions. PMID- 18385801 TI - C677T polymorphism in the methylenetetrahydrofolate reductase gene is associated with primary closed angle glaucoma. AB - PURPOSE: To determine whether or not there is an association of the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism with disease in cohorts of primary open-angle glaucoma (POAG) and primary closed-angle glaucoma (PCAG) from Pakistan. METHODS: This was a prospective study consisting of 150 patients (90 POAG and 60 PCAG) and 70 control subjects. Genomic DNA was extracted from leukocytes of the peripheral blood. MTHFR C677T polymorphism analysis was performed by the polymerase chain reaction-restriction fragment length polymorphism (RFLP) technique. RESULTS: The prevalence of the MTHFR C/T genotype was 22.2% in POAG, 13.3% in PACG, and 18.6% in controls whereas the MTHFR T/T genotype was present solely in the PACG group (6.9%). The difference regarding the T/T genotype between PACG and controls was statistically significant (p<0.01). CONCLUSIONS: The MTHFR C677T polymorphism was found to be associated with PCAG but not POAG in patients of Pakistani origin. PMID- 18385802 TI - A note on the iterative MRI reconstruction from nonuniform k-space data. AB - In magnetic resonance imaging (MRI), methods that use a non-Cartesian grid in k space are becoming increasingly important. In this paper, we use a recently proposed implicit discretisation scheme which generalises the standard approach based on gridding. While the latter succeeds for sufficiently uniform sampling sets and accurate estimated density compensation weights, the implicit method further improves the reconstruction quality when the sampling scheme or the weights are less regular. Both approaches can be solved efficiently with the nonequispaced FFT. Due to several new techniques for the storage of an involved sparse matrix, our examples include also the reconstruction of a large 3D data set. We present four case studies and report on efficient implementation of the related algorithms. PMID- 18385803 TI - Improving image analysis in 2DGE-based redox proteomics by labeling protein carbonyl with fluorescent hydroxylamine. AB - Recent advances in redox proteomics have provided significant insight into the role of oxidative modifications in cellular signalling and metabolism. At present, these techniques rely heavily on Western blots to visualize the oxidative modification and corresponding two dimensional (2D) gels for detection of total protein levels, resulting in the duplication of efforts. A major limitation associated with this methodology includes problematic matching up of gels and blots due to the differences in processing and/or image acquisition. In this study, we present a new method which allows detection of protein oxidation and total protein on the same gel to improve matching in image analysis. Furthermore, the digested protein spots are compatible with standard MALDI mass spectrometry protein identification. The methodology highlighted here may be useful in facilitating the development of biomarkers, assessing potential therapeutic targets and elucidating new mechanisms of redox signalling in redox related conditions. PMID- 18385804 TI - In vivo and in vitro techniques for comparative study of antiviral T-cell responses in the amphibian Xenopus. AB - Activation of lymphocytes in mammals is often quantified by measuring the amount of proliferation during the expansion phase of an immune response. Bromodeoxyuridine (BrdU) incorporation and carboxyfluorescein diacetate succinimidyl ester (CFSE) dilution assays are some of the techniques widely used in mammalian studies of pathogen-induced proliferation and provide a convenient way of quantifying the cellular response. We have extended the use of these proliferation assays to the amphibian Xenopus laevis. We have developed this species as a valuable comparative model to study immunity against a well-known amphibian pathogen, Frog Virus 3 (FV3). Fluorescence activated cell sorting was used to assess the level of BrdU incorporation of lymphocytes in vivo and CFSE dilution in an in vitro activation assay. Both techniques have shown that splenic lymphocytes proliferate specifically upon FV3 challenge. This indicates that common methods for detection of proliferation upon immunologic challenge are easily applied to other vertebrate species, as it highlights the evolutionary conservation of the proliferative nature of immune responses throughout vertebrate phyla. PMID- 18385805 TI - Identification of a calcitriol-regulated Sp-1 site in the promoter of human CD14 using a combined western blotting electrophoresis mobility shift assay (WEMSA). AB - Calcitriol (1alpha, 25-dihydroxyvitamin D(3)) induces the expression of CD14 in mononuclear phagocytes. The mechanisms accounting for this have been unclear since the promoter of CD14 does not contain a canonical vitamin D response element (VDRE). Calcitriol has been shown to regulate the activity of the transcription factor Sp-1 and our analysis of the proximal promoter of CD14 indicated the presence of four Sp-1-like binding sequences. To identify which of these sites might be involved in the response to calcitriol, we used a system incorporating an electrophoretic mobility shift assay (EMSA) coupled to Western blot analysis (WEMSA). Using WEMSA, we found that only one of the Sp-1-like binding sequences, located at position -91 to -79 (relative to the transcription start site), bound the transcription factor Sp1. Sp-1 binding to this site was demonstrable using nuclear extracts from control cells. Notably, binding activity was attenuated in nuclear extracts prepared from cells that had been incubated with calcitriol, thus suggesting Sp-1 involvement in calcitriol induction of CD14 expression. Notably, these results show that like EMSA, WEMSA can be broadly applied to aid in the identification of transcription factors involved in regulating gene expression. WEMSA, however, offers a number of distinct advantages when compared with conventional EMSA. Antibodies used for WEMSA often provide less ambiguous signals than those used in EMSA, and these do not have to recognize epitopes under native conditions. In addition, WEMSA does not require the use of labeled oligos, thus eliminating a significant expense associated with EMSA. PMID- 18385806 TI - Optimized two-dimensional thin layer chromatography to monitor the intracellular concentration of acetyl phosphate and other small phosphorylated molecules. AB - Acetyl phosphate (acetyl-P) serves critical roles in coenzyme A recycling and ATP synthesis. It is the intermediate of the Pta-AckA pathway that inter-converts acetyl-coenzyme A and acetate. Acetyl-P also can act as a global signal by donating its phosphoryl group to specific two-component response regulators. This ability derives from its capacity to store energy in the form of a high-energy phosphate bond. This bond, while critical to its function, also destabilizes acetyl-P in cell extracts. This lability has greatly complicated biochemical analysis, leading in part to widely varying acetyl-P measurements. We therefore developed an optimized protocol based on two-dimensional thin layer chromatography that includes metabolic labeling under aerated conditions and careful examination of the integrity of acetyl-P within extracts. This protocol results in greatly improved reproducibility, and thus permits precise measurements of the intracellular concentration of acetyl-P, as well as that of other small phosphorylated molecules. PMID- 18385807 TI - Recent advances in freeze-fracture electron microscopy: the replica immunolabeling technique. AB - Freeze-fracture electron microscopy is a technique for examining the ultrastructure of rapidly frozen biological samples by transmission electron microscopy. Of a range of approaches to freeze-fracture cytochemistry that have been developed and tried, the most successful is the technique termed freeze fracture replica immunogold labeling (FRIL). In this technique, samples are frozen, fractured and replicated with platinum-carbon as in standard freeze fracture, and then carefully treated with sodium dodecylsulphate to remove all the biological material except a fine layer of molecules attached to the replica itself. Immunogold labeling of these molecules permits their distribution to be seen superimposed upon high resolution planar views of membrane structure. Examples of how this technique has contributed to our understanding of lipid droplet biogenesis and function are discussed. PMID- 18385808 TI - Quantitative evaluation of signaling events in Drosophila S2 cells. AB - Drosophila activates a robust defense response to gram-negative bacteria through the Immune deficiency (Imd) pathway. Imd signaling proceeds through c-Jun N terminal Kinase (JNK), NF-kB and caspase modules. The individual signaling modules act in a highly coordinated manner to yield a stereotypical response to infection. While considerable attention has focused on NF-kB-mediated antimicrobial activities, more recent studies have highlighted the involvement of JNK signaling in the Imd pathway response. JNK signaling occurs in a transitory burst and drives the expression of a number of gene products through the AP-1 transcription factor. In this report, we describe a simple method for the quantification of JNK activation by Western blot analysis or directly in tissue culture plates. PMID- 18385809 TI - Micro-scale flow cytometry-based and biochemical analysis of lipid signaling in primary B cell subpopulations. AB - B cell subpopulations in the spleen have been extensively characterized phenotypically; however, biochemical properties of these cell populations following B cell antigen receptor engagement have not been fully determined due to technical difficulties and limiting cell numbers. We therefore employed mini scale protocols to assess lipid signaling, particularly that of diacylglycerol and inositol trisphosphate, with as few as 0.5x10(6) purified early (T1) and late (T2) transitional B cells. Additionally, utilizing flow cytometric techniques, we determined levels of phosphatidylinositol bisphosphate and calcium mobilization in T1 and T2 cells, as well as mature follicular and marginal zone B cells using less than 1x10(6 )primary B cells. Thus, these biochemical and flow cytometric methodologies can be used to analyse signal-induced changes in phosphatidylinositol bisphosphate levels, diacylglycerol and inositol triphosphate production and calcium in each B cell population. PMID- 18385810 TI - Platelet counts and platelet activation markers in obese subjects. AB - OBJECTIVE: In this work we studied the correlation between platelet count, platelet activation, and systemic inflammation in overweight, obese, and morbidly obese individuals. METHODS AND SUBJECTS: A total of 6319 individuals participated in the study. Complete blood counts, high sensitivity C-reactive protein (hs-CRP) serum levels, and body mass index (BMI) were measured during routine checkups. Platelet activation markers were studied among 30 obese (BMI = 41 +/- 8 kg/m(2)) and 35 nonobese (BMI = 24 +/- 3 kg/m(2)) individuals. Platelet activation status was evaluated by flow cytometry using specific antibodies against the activated platelet membrane glycoprotein IIb/IIIa, p-selectin (CD-62 p), and binding of Annexin-V to platelet anionic phospholipids. RESULTS: Overweight, obese, and morbidly obese females had significantly elevated platelet counts ( P < .0001) compared with normal-weight females. No significant elevation of platelet counts was observed in the male subgroups. A significant age adjusted correlation between BMI and platelet counts ( P < .0001) was found among females. This correlation was attenuated (P = .001) after adjustment for hs-CRP concentrations. The flow cytometry analysis of platelets showed no significant differences in activation marker expression between nonobese and obese individuals. DISCUSSION: Obesity may be associated with elevated platelet counts in females with chronic inflammation. Obesity is not associated with increased platelet activation. PMID- 18385811 TI - COX-2 gene promoter polymorphism and coronary artery disease in middle-aged men: the Helsinki sudden death study. AB - Cyclooxygenase (COX) catalyzes formation of prostaglandins that contribute to the inflammation in atherosclerosis. Our objective was to study whether the functional C variant of the -765G-->C polymorphism in the human COX-2 gene associates with the severity of coronary atherosclerosis measured at the coronary artery level. The Helsinki sudden death study autopsy material (n = 300) comprised of Finnish men who died suddenly. The area of atherosclerotic lesions in the coronary arteries was quantitated, and coronary narrowing was measured. The occurrence of myocardial infarction (MI) was assessed. Genotyping was by restriction endonuclease analysis. Men carrying the minor C allele had larger areas of complicated lesions (P = .024) and a higher number of coronary arteries that had over 50% stenosis (P = .036) compared to men representing the common GG genotype. The COX-2 polymorphism was not associated with MI. Our data suggest that COX-2 may be involved in plaque growth. PMID- 18385812 TI - On the optimum sampling time for the measurement of pollutants in ambient air. AB - No measurement result of any kind is complete without an associated statement of uncertainty. In general, the aim is to minimise this measurement uncertainty to the extent that the measurement made is fit for purpose, or the uncertainty meets an acceptable level such as a "data quality objective". This paper demonstrates how the sampling time used to measure a pollutant in ambient air over a given time period, particularly with automatic instruments, can affect the uncertainty of the measurement result. It is shown that the uncertainty of the overall result depends critically on how the precision characteristics of the measuring instrumentation vary with analyte abundance. An example of the determination of these characteristics for mercury vapour measurement and the conclusions that may be drawn about optimum sampling times to minimise uncertainty are presented. For situations where uncertainty is minimised by the use of long sampling periods, the implications of this strategy on the "information richness" of the data and on the detection of short-term episodes, as well as peak concentrations, are discussed. PMID- 18385813 TI - Spectroscopic OCT by grating-based temporal correlation coupled to optical spectral analysis. AB - Spectroscopic optical coherence tomography (spectroscopic OCT) is an echographic like optical method for biomedical functional imaging. Current spectroscopic optical coherence tomography (OCT) methods rely on a posteriori numerical calculation. We present an alternative for optically accessing the spectroscopic information in OCT, that is, without postprocessing, by using a grating-based correlation and a wavelength demultiplexing system. Spectrally resolved A-scan is directly recorded on the image sensor. Due to the grating-based system, no correlation scan is necessary. The signal is registered in the wavelength-depth plane on a 2D camera that provides a large number of resolved points. In the frame of this paper, we present the principle of the system as well as demonstration results. Advantages and drawback of this system compared to others are discussed. PMID- 18385814 TI - The importance of fever as a predictive symptom for the potency of host's monocytes to release pro- and anti-inflammatory mediators. AB - OBJECTIVE: To clarify whether time lapsing from advent of fever as a first sign of sepsis may be indicative of the potency of monocytes for the release of pro- and anti-inflammatory mediators. METHODS: Monocytes were isolated from blood of 51 septic patients and 9 healthy donors. Monocytes were incubated in the absence and presence of patients' serum and concentrations of tumour necrosis factor alpha (TNF alpha), interleukin (IL)-6, IL-10, and malondialdehyde (MDA) were estimated in supernatants. Patients were divided into three groups: group A: <12 hours; group B: 12-24 hours, and group C: >24 hours between initiation of fever and blood sampling. RESULTS: TNF alpha of supernatants of groups B and C was higher than controls, as also were IL-6 of A and C, IL-10 of A and B, and MDA of A. IL-6 of group A was increased after addition of patients serum. A negative correlation was found between time from initiation of symptoms and IL-6 of monocyte supernatants incubated in the presence of patients serum. Median IL-6 of survivors was higher than nonsurvivors. CONCLUSION: Monocytes are potent for the release of pro- and anti-inflammatory mediators within the first 24 hours upon advent of fever related to sepsis; serum stimulates further release of IL-6 within the first 12 hours. PMID- 18385815 TI - Receptor guided 3D-QSAR: a useful approach for designing of IGF-1R inhibitors. AB - Research by other investigators has established that insulin-like growth factor-1 receptor (IGF-1R) is a key oncological target, and that derivatives of 1, 3 disubstituted-imidazo[1,5-alpha] pyrazine are potent IGF-1R inhibitors. In this paper, we report on our three-dimensional quantitative structure activity relationship (3D-QSAR) studies for this series of compounds. We validated the 3D QSAR models by the comparison of two major alignment schemes, namely, ligand based (LB) and receptor-guided (RG) alignment schemes. The latter scheme yielded better 3D-QSAR models for both comparative molecular field analysis (CoMFA) (q(2) = 0.35, r(2) = 0.95) and comparative molecular similarity indices analysis (CoMSIA) (q(2) = 0.51, r(2) = 0.86). We submit that this might arise from the more accurate inhibitor alignment that results from using the structural information of the active site. We conclude that the receptor-guided 3D-QSAR may be helpful to design more potent IGF-1R inhibitors, as well as to understand their binding affinity with the receptor. PMID- 18385816 TI - Comparison of C-reactive protein and serum amyloid a protein in septic shock patients. AB - Septic shock is a severe inflammatory state caused by an infectious agent. Our purpose was to investigate serum amyloid A (SAA) protein and C-reactive protein (CRP) as inflammatory markers of septic shock patients. Here we evaluate 29 patients in postoperative period, with septic shock, in a prospective study developed in a surgical intensive care unit. All eligible patients were monitored over a 7-day period by sequential organ failure assessment (SOFA) score, daily CRP, SAA, and lactate measurements. CRP and SAA strongly correlated up to the fifth day of observation but were not good predictors of mortality in septic shock. PMID- 18385818 TI - Zinc in human health: effect of zinc on immune cells. AB - Although the essentiality of zinc for plants and animals has been known for many decades, the essentiality of zinc for humans was recognized only 40 years ago in the Middle East. The zinc-deficient patients had severe immune dysfunctions, inasmuch as they died of intercurrent infections by the time they were 25 years of age. In our studies in an experimental human model of zinc deficiency, we documented decreased serum testosterone level, oligospermia, severe immune dysfunctions mainly affecting T helper cells, hyperammonemia, neurosensory disorders, and decreased lean body mass. It appears that zinc deficiency is prevalent in the developing world and as many as two billion subjects may be growth retarded due to zinc deficiency. Besides growth retardation and immune dysfunctions, cognitive impairment due to zinc deficiency also has been reported recently. Our studies in the cell culture models showed that the activation of many zinc-dependent enzymes and transcription factors were adversely affected due to zinc deficiency. In HUT-78 (T helper 0 [Th(0)] cell line), we showed that a decrease in gene expression of interleukin-2 (IL-2) and IL-2 receptor alpha(IL 2Ralpha) were due to decreased activation of nuclear factor-kappaB (NF-kappaB) in zinc deficient cells. Decreased NF-kappaB activation in HUT-78 due to zinc deficiency was due to decreased binding of NF-kappaB to DNA, decreased level of NF-kappaB p105 (the precursor of NF-kappaB p50) mRNA, decreased kappaB inhibitory protein (IkappaB) phosphorylation, and decreased Ikappa kappa. These effects of zinc were cell specific. Zinc also is an antioxidant and has anti-inflammatory actions. The therapeutic roles of zinc in acute infantile diarrhea, acrodermatitis enteropathica, prevention of blindness in patients with age related macular degeneration, and treatment of common cold with zinc have been reported. In HL-60 cells (promyelocytic leukemia cell line), zinc enhances the up regulation of A20 mRNA, which, via TRAF pathway, decreases NF-kappaB activation, leading to decreased gene expression and generation of tumor necrosis factor alpha (TNF-alpha), IL-1beta, and IL-8. We have reported recently that in both young adults and elderly subjects, zinc supplementation decreased oxidative stress markers and generation of inflammatory cytokines. PMID- 18385819 TI - The IGF-1 receptor as a therapeutic target to improve endothelial progenitor cell function. PMID- 18385817 TI - Host defense peptides in wound healing. AB - Host defense peptides are effector molecules of the innate immune system. They show broad antimicrobial action against gram-positive and -negative bacteria, and they likely play a key role in activating and mediating the innate as well as adaptive immune response in infection and inflammation. These features make them of high interest for wound healing research. Non-healing and infected wounds are a major problem in patient care and health care spending. Increasing infection rates, growing bacterial resistance to common antibiotics, and the lack of effective therapeutic options for the treatment of problematic wounds emphasize the need for new approaches in therapy and pathophysiologic understanding. This review focuses on the current knowledge of host defense peptides affecting wound healing and infection. We discuss the current data and highlight the potential future developments in this field of research. PMID- 18385820 TI - Detection of gene interactions based on syntactic relations. AB - Interactions between proteins and genes are considered essential in the description of biomolecular phenomena, and networks of interactions are applied in a system's biology approach. Recently, many studies have sought to extract information from biomolecular text using natural language processing technology. Previous studies have asserted that linguistic information is useful for improving the detection of gene interactions. In particular, syntactic relations among linguistic information are good for detecting gene interactions. However, previous systems give a reasonably good precision but poor recall. To improve recall without sacrificing precision, this paper proposes a three-phase method for detecting gene interactions based on syntactic relations. In the first phase, we retrieve syntactic encapsulation categories for each candidate agent and target. In the second phase, we construct a verb list that indicates the nature of the interaction between pairs of genes. In the last phase, we determine direction rules to detect which of two genes is the agent or target. Even without biomolecular knowledge, our method performs reasonably well using a small training dataset. While the first phase contributes to improve recall, the second and third phases contribute to improve precision. In the experimental results using ICML 05 Workshop on Learning Language in Logic (LLL05) data, our proposed method gave an F-measure of 67.2% for the test data, significantly outperforming previous methods. We also describe the contribution of each phase to the performance. PMID- 18385821 TI - An algorithm for finding functional modules and protein complexes in protein protein interaction networks. AB - Biological processes are often performed by a group of proteins rather than by individual proteins, and proteins in a same biological group form a densely connected subgraph in a protein-protein interaction network. Therefore, finding a densely connected subgraph provides useful information to predict the function or protein complex of uncharacterized proteins in the highly connected subgraph. We have developed an efficient algorithm and program for finding cliques and near cliques in a protein-protein interaction network. Analysis of the interaction network of yeast proteins using the algorithm demonstrates that 59% of the near cliques identified by our algorithm have at least one function shared by all the proteins within a near-clique, and that 56% of the near-cliques show a good agreement with the experimentally determined protein complexes catalogued in MIPS. PMID- 18385822 TI - Proton pump inhibitors and gastritis. AB - Proton pump inhibitors (PPIs) are novel compounds that strongly inhibit the H(+)/K(+)-ATPase in the gastric parietal cells to cause profound suppression of acid secretion. Acid-generating ATPase, also known as vacuolar-type ATPase, is located in the lysozomes of leukocytes and osteoclasts and its activity is also reportedly influenced by treatment with PPIs. This concept is supported by the results of studies using autoradiography in which (3)H-Lansoprazole uptake sites were clearly detected in the cytoplasmic granules of neutrophils infiltrating the gastric mucosa. In vitro studies indicate that PPIs increase the intra-vacuolar pH in the lysosomes of purified neutrophils and attenuate the adherence of neutrophils to the vascular endothelium. In clinical practice, the acidic environment in the stomach plays a critical role in the development of gastritis induced by Helicobacter pylori (H. pylori). This is worthy of note, because persistent gastritis often results in atrophic and metaplastic changes in the gastric mucosa, which are believed to be preneoplastic abnormalities. In patients with H. pylori-infection, PPI therapy causes corpus-predominant gastritis, which is frequently found in the background mucosa in patients with gastric cancer. The efficacy and safety of long-term PPI-treatment have not been conclusive, thus we need to pay more attention to the additional pharmacological actions of PPIs. PMID- 18385823 TI - Introduction to serial review: heme oxygenase and carbon monoxide: medicinal chemistry and biological effects. PMID- 18385824 TI - Application of heme oxygenase-1, carbon monoxide and biliverdin for the prevention of intestinal ischemia/reperfusion injury. AB - Intestinal ischemia/reperfusion (I/R) injury occurs frequently in a variety of clinical settings, including mesenteric artery occlusion, abdominal aneurism surgery, trauma, shock, and small intestinal transplantation, and is associated with substantial morbidity and mortality. Although the exact mechanisms involved in the pathogenesis of intestinal I/R injury have not been fully elucidated, it is generally believed that polymorphonuclear neutrophils, pro-inflammatory cytokines, and mediators generated in the setting of oxidative stress, such as reactive oxygen species (ROS), play important roles. Heme oxygenase (HO) is the rate-limiting enzyme that catalyzes the degradation of heme into equimolar quantities of biliverdin and carbon monoxide (CO), while the central iron is released. An inducible form of HO (HO-1), biliverdin, and CO, have been shown to possess generalized endogenous anti-inflammatory activities and provide protection against intestinal I/R injury. Further, recent observations have demonstrated that exogenous HO-1 expression, as well as exogenously administered CO and biliverdin, have potent cytoprotective effects on intestinal I/R injury as well. Here, we summarize the currently available data regarding the role of the HO system in the prevention intestinal I/R injury. PMID- 18385825 TI - Effects on the Human Body of a Dietary Supplement Containing L-Carnitine and Garcinia cambogia Extract: A Study using Double-blind Tests. AB - The effect of a dietary supplement with L-carnitine (600 mg/day) and Garcinia cambogia extract (500 mg/day as hydroxycitric acid) as main ingredients was studied in 35 healthy volunteers {48.3 +/- 6.9 years, body mass index (BMI): 26.3 +/- 1.7} in a double-blind test (18 subjects in the Test Group and 17 in the Control Group). The yearly examination includes the standard yearly medical tests done in Japan, tests for assessing hormonal age, and a survey for assessing physical and mental fitness of the subjects, called the Anti-Aging QOL Common Questionnaire (AAQol). Use of this supplement significantly improved the level of lipid peroxides (-12.8%) in the blood as well as physical symptoms such as "tired eyes," "blurry eyes," "muscle pain/stiffness," "early satiety," "epigastralgia," "dizziness," "arthralgia" and "easily breaking into a sweat." The Control Group showed a significantly favorable improvement rate, especially for "dizziness." On the other hand, groups of subjects using the test compounds saw a significant rise in total cholesterol (4.5%), fasting blood sugar (4.1%) and HbA1c (3.4%). Our findings suggest that the consumption of the supplement can reduce the oxidative damage; however, the effect on QOL was equivocal. Garcinia cambogia extract did not show dietary efficacy. PMID- 18385826 TI - A Genetic Variant of the CD14 C-159T in Patients with Functional Dyspepsia (FD) in Japanese Subjects. AB - Inflammatory changes in the gastric mucosa are commonly observed in Japanese patients with functional dyspepsia (FD). However, detailed data regarding the relationship between the genetic regulatory factors of inflammation and FD are not available. CD14 is an important mediator of the inflammatory response in the first line of host defense by recognition of Lipopolysaccharide (LPS). We aimed to investigate the association between CD14 promoter C-159T polymorphism and FD in a Japanese population. 108 patients with FD and 99 non-dyspeptic subjects enrolled in this study. Dyspeptic symptoms were divided according to Rome III criteria. CD14 gene C-159T polymorphism was determined by polymerase chain reaction-restriction fragment length polymorphism. In the non-dyspeptics, the CD14 genotype distribution was 28CC (28.3%), 51CT (51.5%), 21TT (21.2%). Meanwhile, the CD14 genotype distribution in FD was 31CC (28.4%), 56CT (51.4%), 22TT (20.2%). The genotype distribution was not significantly different. There was no significant difference between two groups in the genotype distribution. We did not found any association between CD14 genotypes and dyspeptic patients in different gender and Helicobacter pylori infection status. No significant association was also found between CD14 polymorphism and any of different subtypes of FD according to Rome III while there was a weak correlation between TT genotype and PDS in male subjects (TT vs others, OR = 3.18, 95% CI = 0.98 10.26, p = 0.06). In conclusion, our results suggest that CD14 polymorphism is unlikely to associate with susceptibility of dyspeptic symptoms. The role of inflammation related-gene polymorphisms to the development of dyspepsia needs to further evaluation. PMID- 18385827 TI - Role of plasma protein and low-molecular weight substances in the change of hydroxyl radical scavenging activity in hemodialysis patients. AB - While it is well known that hemodialysis (HD) patients with end stage renal failure are exposed to high oxidative stress, there is not a general opinion regarding whether antioxidant activity is high or low in these patients. We evaluated the variation of plasma hydroxyl radical scavenging activity (p-HRSA) by a single-session of HD in 69 patients by using a new system, reactive flow injection electron spin resonance. And then comparing p-HRSA with their biochemical parameters, we tried to elucidate what components affected p-HRSA in the HD patients. The average of p-HRSA significantly increased after HD and the variation of p-HRSA by HD was correlated with that of plasma total protein (TP). In 5 patients however, their p-HRSA decreased after HD, in spite of increasing TP. In pre-HD, the p-HRSA values and hydroxyl radical scavenging activity of low molecular weight fraction of plasma were significantly higher in these 5 patients than in patients whose p-HRSA increased after HD. These 5 patients were in an inflammatory state. These findings suggest that p-HRSA is mainly affected by TP, but caution should be exercised in patients who have high p-HRSA before HD and whose p-HRSA does not increase after HD. PMID- 18385828 TI - Octacosanol attenuates disrupted hepatic reactive oxygen species metabolism associated with acute liver injury progression in rats intoxicated with carbon tetrachloride. AB - We examined whether octacosanol, the main component of policosanol, attenuates disrupted hepatic reactive oxygen species metabolism associated with acute liver injury progression in rats intoxicated with carbon tetrachloride (CCl(4)). In rats intoxicated with CCl(4) (1 ml/kg, i.p.), the activities of serum transaminases increased 6 h after intoxication and further increased at 24 h. In the liver of CCl(4)-intoxicated rats, increases in lipid peroxide (LPO) concentration and myeloperoxidase activity and decreases in superoxixde dismutase activity and reduced glutathione (GSH) concentration occurred 6 h after intoxication and these changes were enhanced with an increase in xanthine oxidase activity and a decrease in catalase activity at 24 h. Octacosanol (10, 50 or 100 mg/kg) administered orally to CCl(4)-intoxicated rats at 6 h after intoxication attenuated the increased activities of serum transaminases and the increased hepatic myeloperoxidase and xanthine oxidase activities and LPO concentration and the decreased hepatic superoxide dismutase and catalase activities and GSH concentration found at 24 h after intoxication dose-dependently. Octacosanol (50 or 100 mg/kg) administered to untreated rats decreased the hepatic LPO concentration and increased the hepatic GSH concentration. These results indicate that octacosanol attenuates disrupted hepatic reactive oxygen species metabolism associated with acute liver injury progression in CCl(4)-intoxicated rats. PMID- 18385829 TI - Enhanced expressions of endothelin-converting enzyme and endothelin receptors in human colonic tissues of Crohn's disease. AB - Endothelin-1, a powerful vasoconstrictor, forms the endothelin system together with endothelin-converting enzyme and endothelin type A and type B receptors. These endothelin system components are considered to participate in inflammatory and wound healing responses. Previous reports have suggested a role for the endothelin-1 in the pathology of Crohn's disease. In the present study, we immunohistochemically investigated the expressions of the endothelin system components in affected human intestinal tissues of Crohn's disease. Eighteen surgical specimens of colonic tissue obtained from patients with Crohn's disease and 12 normal colonic tissues as controls were examined. Frozen tissue sections cut from the samples were subjected to the immunohistochemical single and double staining. The endothelin system components were expressed mainly in the muscular layers and blood vessels. In diseased colonic tissues, inflammatory infiltration and fibrotic tissue reactions with marked smooth muscle cell proliferation were frequently seen, and were closely associated with increased expressions of the endothelin system components. These results strongly suggest that endothelin converting enzyme and endothelin type A and type B receptors collectively play a role in the inflammatory and fibrogenic processes of Crohn's disease. Especially, submucosal smooth muscle proliferation, a histological hallmark of strictures, may be attributable to the upregulated endothelin system. PMID- 18385830 TI - Association between Total Antioxidant Capacity in Breast Milk and Postnatal Age in Days in Premature Infants. AB - This study aimed to consider the significance of breast milk in preventing oxidative stress by comparing total antioxidant capacity (TAC) in breast milk and formula milk for premature infants, demonstrating the relationship between TAC in breast milk and postnatal age in days. We used the biological anti-oxidant potential test, a new method to measure TAC in breast milk. Breast milk for premature infants were stored at -20 degrees C and thawed within 48 h of collection. We measured TAC in two types of formula milk in the same way. TAC was clearly higher in breast milk than formula milk. Although a negative correlation was observed between TAC in breast milk and age when collected, TAC was always higher than the average TAC in formula milk. TAC in breast milk is higher than TAC in formula milk. We suggest the importance of breast milk for preventing oxidative stress and starting breastfeeding early. PMID- 18385831 TI - Stimulatory Effects of CO(2) Laser, Er:YAG Laser and Ga-Al-As Laser on Exposed Dentinal Tubule Orifices. AB - We investigated the effects of lasers irradiation on the exposed dentinal tubule. Human tooth specimens with exposed dentinal tubule orifices were used. Three types of lasers (CO(2) laser, Er:YAG laser and Ga-Al-As laser) were employed. The parameters were 1.0 W in continuous-wave mode with an irradiation time of 30 s for the CO(2) laser, 30 mJ in continuous-wave mode with an irradiation time of 60 s for the Er:YAG laser, and 1.0 W in continuous-wave mode with an irradiation time of 60 s for the Ga-Al-As laser. A non-irradiated group was used as a control. After laser irradiation, the dentinal surface of each sample was observed using SEM. Afterwards, all samples were immersed in methylene blue dye solution in order to evaluate the penetration of the dye solution and observe the change in dentinal permeability after laser irradiation. SEM observation showed that the control group had numerous exposed dentinal tubule orifices, whereas these orifices were closed in the laser-irradiated groups. There was consistent dye penetration into the pulp chamber in the control group, whereas no dye penetration was evident in the laser-irradiated groups. Therefore, laser appears to be a promising treatment for reducing permeation through exposed dentinal tubules. PMID- 18385832 TI - Long-term Ultrasonographic Follow-up Study of Gastric Motility in Patients with Functional Dyspepsia. AB - Although patients with functional dyspepsia complain of epigastric symptoms, the relation between these symptoms and gastric motility remains controversial. There are few reports on the clinical course of functional dyspepsia, including changes in gastric motility, observed over a considerably long period. We conducted a study to examine association between changes in symptoms and changes in ultrasonographically evaluated gastric motility over a long-term follow-up period in patients with functional dyspepsia. Forty patients (18 men, 22 women; mean age, 53.7 years) with functional dyspepsia were followed up by medical interview, physical examination, endoscopy, and ultrasonography for gastric motility. Follow up ranged from 1.0 to 7.8 years (mean, 3.0 years). Ultrasonographic evaluation of gastric motility included gastric emptying rate and antral contractions. During the follow-up period, patients were treated with proton pump inhibitors, H2 blockers, or prokinetics. Symptoms improved in 21 patients (group A), but symptoms persisted or worsened in 19 patients (group B). There were no significant differences in clinical characteristics between the two groups. Gastric motility improved in group A but not in group B. In conclusion, improved gastric motility appears to correspond to and may explain improved symptoms in some patients with functional dyspepsia. PMID- 18385833 TI - A Mouse Model of Metabolic Syndrome; Increase in Visceral Adipose Tissue Precedes the Development of Fatty Liver and Insulin Resistance in High-Fat Diet-Fed Male KK/Ta Mice. AB - To determine the relative contribution of obesity and visceral white adipose tissue (WAT) to metabolic syndrome, we developed a model that is susceptible to high-fat diet-induced obesity and insulin resistance using male KK/Ta mice. The ratio of WAT weight to body weight was greater in the high-fat diet group compared with the control group in 10-, 14-, and 22-week-old mice. The increase in visceral WAT preceded development of fatty liver and insulin resistance. Adiponectin mRNA expression in WAT was markedly decreased before the decrease in its plasma levels or the development of insulin resistance. Insulin resistance appeared in association with fatty infiltration and TNF-alpha expression in the liver in 22-week-old mice. These data indicate that our mouse model would be useful for future studies that investigate the role of visceral WAT and its products in the development of metabolic syndrome. PMID- 18385834 TI - Bofutsushosan, an Oriental Herbal Medicine, Attenuates the Weight Gain of White Adipose Tissue and the Increased Size of Adipocytes Associated with the Increase in Their Expression of Uncoupling Protein 1 in High-Fat Diet-Fed Male KK/Ta mice. AB - Bofutsushosan (BOF), an oriental herbal medicine, has been used as an anti obesity drug in overweight patients. In the present study, to evaluate the anti obesity and anti-diabetic effects of BOF, we investigated the effects of BOF on the white adipose tissue (WAT) weight, the size of adipocytes, adiponectin expression, and oral glucose tolerance test results in high-fat diet-fed male KK/Ta mice. In addition, the mRNA expression levels of uncoupling protein 1 (UCP1) and UCP2 mRNA in WAT and brown adipose tissue (BAT) were measured. 6-week old KK/Ta mice were divided into four groups and fed a purified powdered basal diet (the BD group), a purified high-fat (HF) powdered diet containing suet powder at 37.5 g/100 g diet (the HF group), a high-fat diet plus 1.0% bofutsushosan (BOF) treatment (the HF + BOF group), or a high-fat diet plus 1.0% daisaikoto (DAI) treatment (the HF + DAI group) for 4 weeks. The weight of WAT and the size of adipocytes were increased in the HF group compared with those in the BD group, and these increases in the HF group were significantly inhibited in the HF + BOF group, but not affected in the HF + DAI group. There were no statistically significant differences in plasma levels and tissue mRNA levels of adiponectin among the four groups. There were no significant differences in UCP1 mRNA expression of BAT among the four groups. The expression of UCP1 mRNA in WAT was found in the HF + BOF group, but little expression was seen in the WAT of the BD, HF, or HF + DAI groups. The elevated plasma glucose levels and responses after the glucose loading in the HF group tended to decrease in the HF + BOF group. These results suggest that BOF decreases the weight and size gains of WAT along with up-regulating UCP1 mRNA in WAT in high-fat diet-fed mice. PMID- 18385835 TI - Saposin B is a human coenzyme q10-binding/transfer protein. AB - Coenzyme Q10 (CoQ10) is essential for ATP production in the mitochondria, and is an important antioxidant in every biomembrane and lipoprotein. Due to its hydrophobicity, a binding and transfer protein for CoQ10 is plausible, but none have yet been isolated and characterized. Here we purified a CoQ10-binding protein from human urine and identified it to be saposin B, a housekeeping protein necessary for sphingolipid hydrolysis in lysosomes. We confirmed that cellular saposin B binds CoQ10 in human sperm and the hepatoma cell line HepG2 by using saposin B monoclonal antibody. The molar ratios of CoQ10 to saposin B were estimated to be 0.22 in urine, 0.003 in HepG2, and 0.12 in sperm. We then confirmed that aqueous saposin B extracts CoQ10 from hexane to form a saposin B CoQ10 complex. Lipid binding affinity to saposin B decreased in the following order: CoQ10>CoQ9>CoQ7>>alpha-tocopherol>>cholesterol (no binding). The CoQ10 binding affinity to saposin B increased with pH, with maximal binding seen at pH 7.4. On the other hand, the CoQ10-donating activity of the saposin B-CoQ10 complex to erythrocyte ghost membranes increased with decreasing pH. These results suggest that saposin B binds and transports CoQ10 in human cells. PMID- 18385836 TI - Recent advances in enantioselective [2 + 2 + 2] cycloaddition. AB - Enantioselective cycloaddition using chiral transition metal catalysts is an atom economical and efficient synthetic tool for the construction of chiral carbo- and heterocyclic skeletons. This short account discloses our recent results of inter- and intramolecular enantioselective [2 + 2 + 2] cycloadditions of alkyne and/or alkene moiety(ies). Chiral iridium complexes catalyzed the alkyne trimerization for the generation of axial chirality(ies), and chiral rhodium ones catalyzed alkyne-alkyne-alkene cyclization for the generation of a quaternary carbon including spirocyclic system. PMID- 18385837 TI - Nucleoside, nucleotide and oligonucleotide based amphiphiles: a successful marriage of nucleic acids with lipids. AB - Amphiphilic molecules based on nucleosides, nucleotides and oligonucleotides are finding more and more biotechnological applications. This Perspective highlights their synthesis, supramolecular organization as well as their applications in the field of biotechnology. PMID- 18385839 TI - Urea derivatives are highly active catalysts for the base-mediated generation of terminal epoxides from aldehydes and trimethylsulfonium iodide. AB - N,N'-Diarylureas have been shown to efficiently catalyse sulfonium ylide-mediated aldehyde epoxidation reactions for the first time. These processes are of broad scope and can be coupled with a subsequent Cu(II) ion-catalysed Meinwald rearrangement to give an efficient and convenient protocol for aldehyde homologation without intermediate purification. PMID- 18385838 TI - A highly fluorescent nucleoside analog based on thieno[3,4-d]pyrimidine senses mismatched pairing. AB - A highly emissive nucleobase analog, based on a thieno[3,4-d]pyrimidine core, is enzymatically incorporated into RNA oilgonucleotides that function as base discriminating fluorescent probes. PMID- 18385840 TI - A novel palladium-catalyzed hydroalkoxylation of alkenes with a migration of double bond. AB - A novel palladium-catalyzed addition of alcohols to olefins was developed, in which a migration of double bond was involved. By this new method, a variety of allylic ethers were prepared with moderate to high yields under mild conditions. PMID- 18385841 TI - N-->O-Acyl shift in Fmoc-based synthesis of phosphopeptides. AB - Synthetic phosphopeptides are frequently used as chemical probes to explore protein-protein interactions involved in cellular signal transduction. Most commonly, the solid-phase synthesis of phosphotyrosine-containing peptides is performed by applying the Fmoc-strategy and N-Fmoc-protected tyrosine derivatives bearing acid-labile phospho protecting groups. We observed a side-reaction, the isomerisation at threonine, which furnishes depsipeptides. It is shown that the rate of N-->O-acyl migration depends on the sequence context. Depsipeptides were formed most rapidly when the phosphotyrosine was located in the +2 position. Furthermore, different phosphotyrosine building blocks were compared and a suitable method that provides phosphopeptides in enhanced purity and yield is suggested. PMID- 18385842 TI - Inhibition and dispersion of Pseudomonas aeruginosa biofilms with reverse amide 2 aminoimidazole oroidin analogues. AB - The marine alkaloid oroidin along with a small library of reverse amide (RA) 2 aminoimidazoles were synthesized and assayed for anti-biofilm activity against PAO1 and PA14, two strains of the medically relevant gamma-proteobacterium Pseudomonas aeruginosa. Analogues that contained a long, linear alkyl chain were more potent inhibitors than the natural product at preventing the formation of PAO1 and PA14 biofilms. The most active compound in the series was also shown to disperse established PAO1 and PA14 biofilms at low micromolar concentrations. PMID- 18385843 TI - Solvent-dependent oxidations of 5- and 6-azaindoles to trioxopyrrolopyridines and functionalised azaindoles. AB - A regioselective synthesis of 4,7-dimethoxy 5- and 6-azaindoles 2 has been achieved, based on the appropriate choice of ortho-directing or ortho-repulsing groups in the formylation of a pyridine ring. Studies on the regioselectivity of the formylation step and on the preparation of azidoacrylate intermediates 4 are described in this paper. The reactivity of the 5- and 6-azaindole structures towards BBr3-mediated selective monodemethylation and oxidative demethylation reactions were also investigated. The regioselectivity of the deprotection was confirmed using a chemical approach. Oxidation reactions were then carried out on either dimethoxy- or hydroxymethoxyazaindoles, in different solvents, using [bis(trifluoroacetoxy)iodo]benzene. In acetonitrile-water, trioxopyrrolopyridines 12 were obtained, whereas the formation of functionalised azaindoles 17 was observed in acetonitrile-methanol. The tautomeric structure of the trioxopyrrolopyridines was proved by X-ray diffraction analysis. PMID- 18385844 TI - Direct solid-phase synthesis and fluorescence labeling of large, monodisperse mannosylated dendrons in a peptide synthesizer. AB - Fluorophore-labeled glycodendrimers have potential use in the study of carbohydrate-protein interactions by fluorescence spectroscopy and imaging methods. The current solution-phase methods for preparation of such glycoconjugates are labour intensive. On the other hand, the intrinsically more efficient solid-phase methods have been explored only at low generations. Herein we disclose a direct, expedient glycodendrimer synthesis from commercially available or easily prepared building blocks by machine-assisted solid-phase peptide synthesis (SPPS). Large, monodisperse 4th- and 5th-generation polylysine dendrons are prepared and capped with 16 and 32 mannose residues, respectively, in a single synthetic operation. Incorporation of a C-terminal lysine residue in the 4th-generation dendron allows fluorescence labelling with a number of common labels on resin, in organic solvent or in aqueous buffer, as required. A single HPLC purification is sufficient in all cases to obtain a homogeneous sample. The monodispersity of the glycodendrons is confirmed by MALDI-TOF. FITC-labeled 4th generation glycodendron is an excellent probe for the imaging studies of mannose receptor-mediated entry of into dendritic cells by confocal fluorescence microscopy. PMID- 18385845 TI - Cyclic tetrapeptides via the ring contraction strategy: chemical techniques useful for their identification. AB - Cyclic tetrapeptides are a class of natural products that have been shown to have broad ranging biological activities and good pharmacokinetic properties. In order to synthesise these highly strained compounds a ring contraction strategy had previously been reported. This strategy was further optimised and a suite of techniques, including the Edman degradation and mass spectrometry/mass spectrometry, were developed to enable characterisation of cyclic tetrapeptide isomers. An NMR solution structure of a cyclic tetrapeptide was also generated. To illustrate the success of this strategy a library of cyclic tetrapeptides was synthesised. PMID- 18385846 TI - Synthesis of sialoclusters appended to calix[4]arene platforms via multiple azide alkyne cycloaddition. New inhibitors of hemagglutination and cytopathic effect mediated by BK and influenza A viruses. AB - Tetra- and octavalent sialoside clusters were prepared in good yields exploiting for the first time the multiple copper-catalyzed cycloaddition of a propargyl thiosialoside with calix[4]arene polyazides. The cycloadducts featured the hydrolytically stable carbon-sulfur bond at the anomeric position and the 1,4 disubstituted triazole ring as the spacer between the sialic acid moieties and the platform. It was demonstrated that these unnatural motifs did not hamper the desired biological activity of the sialoclusters. In fact, they were able to inhibit, at submillimolar concentrations, the hemagglutination and the viral infectivity mediated both by BK and influenza A viruses. PMID- 18385847 TI - Synthesis of carboxylic amides by ring-opening of oxazolidinones with Grignard reagents. AB - Treatment of N-alkyl-oxazolidin-2-ones with Grignard reagents gives tertiary carboxylic amide products. Various substituted oxazolidinones can be used as illustrated with phenyl, benzyl or isopropyl groups on the 4-position, and methyl, benzyl or p-methoxybenzyl groups on the 3-position (the nitrogen atom). A selection of Grignard reagents were successful, including allyl, benzyl, alkyl and phenyl magnesium halides. The organomagnesium species attacks the carbonyl group and promotes ring-opening of the oxazolidinone. The product tertiary amides are useful substrates for stereoselective transformations and were applied to a highly selective enolate alkylation and to a ring-closing metathesis reaction to a six-membered lactam and hence a formal synthesis of the alkaloids (-)-coniine and (+)-stenusine. PMID- 18385848 TI - Synthesis and bioactivity of the gibberellin, 18-hydroxy-GA1 (GA132). AB - As part of a study to confirm putative structural assignments to new gibberellins and to furnish sufficient quantities for biological investigations, a twenty step synthesis of 18-hydroxy GA1 from gibberellic acid (GA3) is described, allowing the confirmation of structure for a new gibberellin, GA132, that occurs in developing grains of barley (Hordeum vulgare). The early part of the sequence involved cleavage of the C(3)-C(4) bond in the A-ring of a 3-oxo intermediate. The ring was then reformed as part of a "domino" process involving the conjugate addition of alkoxide to an alpha-methylene lactone moiety followed by an intramolecular aldol reaction. The bioactivities of the new GA, and its 18 hydroxy-GA4 relative, have been confirmed in dwarf barley growth and alpha amylase induction assays. PMID- 18385849 TI - Synthesis of multivalent Streptococcus suis adhesion inhibitors by enzymatic cleavage of polygalacturonic acid and 'click' conjugation. AB - A galabiose disaccharide building block was synthesized by an efficient pectinase cleavage of polygalacturonic acid and subsequent chemical functional group transformations. Besides the disaccharide, the corresponding trisaccharide was also obtained and modified. The compounds were subsequently conjugated to dendrimers with up to eight end groups using 'click' chemistry. The compounds were evaluated as inhibitors of adhesion of the pathogen Streptococcus suis in a hemagglutination assay and strong inhibition was observed for the tetra- and octavalent galabiose compound with MIC values in the low nanomolar range. The corresponding octavalent trisaccharide was a ca. 20-fold weaker inhibitor. PMID- 18385850 TI - Kinetic resolution of donor-functionalised tertiary alcohols by Cu-H-catalysed stereoselective silylation using a strained silicon-stereogenic silane. AB - A series of propargylic tertiary alcohols decorated with an sp2-hybridised nitrogen donor were kinetically resolved by reagent-controlled dehydrogenative Si O coupling with a strained, highly reactive silicon-stereogenic cyclic silane. PMID- 18385851 TI - Stereoselective tris-glycosylation to introduce beta-(1-->3)-branches into gentiotetraose for the concise synthesis of phytoalexin-elicitor heptaglucoside. AB - Dodecyl thioglycosides (3, 4, 5) were prepared by conventional transformation of d-glucose and used as new glycosyl donors for a short-step synthesis of phytoalexin elicitor heptaglucoside. A gentio-tetraoside derivative (6) having three hydroxyl groups was synthesized by NIS-TfOH promoted glycosylate in more than 90% yield followed by selective removal of temporary protective groups. Undesired formation of alpha-glycosides at the introduction of beta-(1-->3) branches into gentio-oligosaccharides was found to be suppressed by use of a thiophilic reagent system, BSP (1-benzenesulfinyl piperidine)-Tf2O, giving the heptaglucoside in only four glycosylation steps. PMID- 18385852 TI - 2'-Deoxyimmunosine: stereoselective synthesis, base pairing and duplex stability of oligonucleotides containing 8-oxo-7-thiaguanine. AB - Oligonucleotides containing 7-thia-8-oxoguanine represent a new class of molecules in which sulfur replaces the 7-nitrogen of a purine base. The monomeric 7-thia-8-oxoguanine 2'-deoxyribonucleoside (2'-deoxyimmunosine, 4) was prepared by nucleobase anion glycosylation in a regio- and stereoselective way employing 5 {[(di-n-butylamino)methylidene]amino}thiazolo[4,5-d]pyrimidine-2,7(3H,6H)-dione (18) and 1-chloro-2-deoxy-3,5-di-O-p-toluoyl-alpha-d-erythro-pentofuranose (6). The nucleoside was converted into the phosphoramidite and oligonucleotides were prepared by solid-phase synthesis. Oligonucleotide duplexes containing the 4-dC base pair show a similar stability as those containing the dG-dC motif. Thus the sterically demanding sulfur and the additional 8-oxo group are well accommodated in the major groove of DNA. As expected, compound 4 does not form a Hoogsteen pair, as reported for 8-oxo-2'-deoxyguanosine. Compared to 2'-deoxyguanosine, 2' deoxyimmunosine shows a better mismatch discrimination in Watson-Crick base pairs. PMID- 18385853 TI - Chemical and biomimetic total syntheses of natural and engineered MCoTI cyclotides. AB - The naturally-occurring cyclic cystine-knot microprotein trypsin inhibitors MCoTI I and MCoTI-II have been synthesised using both thia-zip native chemical ligation and a biomimetic strategy featuring chemoenzymatic cyclisation by an immobilised protease. Engineered analogues have been produced containing a range of substitutions at the P1 position that redirect specificity towards alternative protease targets whilst retaining excellent to moderate affinity. Furthermore, we report an MCoTI analogue that is a selective low-microM inhibitor of foot-and mouth-disease virus (FMDV) 3C protease, the first reported peptide-based inhibitor of this important viral enzyme. PMID- 18385854 TI - Synthesis of mono- and 1,3-disubstituted allenes from propargylic amines via palladium-catalysed hydride-transfer reaction. AB - Mono- and 1,3-disubstituted allenes were synthesized from the corresponding propargylamines via palladium-catalysed hydride-transfer reaction. In the current transformation, propargylic amines can be handled as allenyl anion equivalents and introduced into various electrophiles to be transformed into allenes under palladium-catalyzed conditions. PMID- 18385855 TI - Total synthesis of (-)-ulapualide A, a novel tris-oxazole macrolide from marine nudibranchs, based on some biosynthesis speculation. AB - A new, second generation, total synthesis of ulapualide A (1), whose stereochemistry was recently determined from X-ray analysis of its complex with the protein actin, is described. The synthesis is designed and based on some speculation of the biosynthetic origin of the contiguous tris-oxazole unit in ulapualide A, alongside that of the related co-metabolites that contain only two oxazole rings, e.g. 6 and 7. The mono-oxazole carboxylic acid 67b and the mono oxazole secondary 55b alcohol which, together, contain all of the 10 asymmetric centres in the natural metabolite, were first elaborated using a combination of contemporary asymmetric synthesis protocols. Esterification of 67b with 55b under Yamaguchi conditions gave the ester 77 which was then converted into the omega amino acid 18a following simultaneous deprotection of the t-butyl ester and the N Boc protecting groups. Macrolactamisation of 18a, using HATU, now gave the key intermediate macrolactam 17, containing two of the three oxazole rings in ulapualide A (1). A number of procedures were used to introduce the third oxazole ring in ulapualide A from 17, including: a) cyclodehydration to the oxazoline 78a followed by oxidation using nickel peroxide leading to 76; b) dehydration to the enamide 79, followed by conversion into the methoxyoxazoline 78b, via 80, and elimination of methanol from 78b using camphorsulfonic acid. The tris-oxazole macrolide 76 was next converted into the aldehyde 82b in four straightforward steps, which was then reacted with N-methylformamide, leading to the E alkenylformamide 83. Removal of the TBDPS protection at C3 in 83 finally gave (-) ulapualide A, whose 1H and 13C NMR spectroscopic data were indistinguishable from those obtained for naturally derived material. It is likely that the tris-oxazole unit in ulapualide A (1) is derived in nature from a cascade of cyclodehydrations from an acylated tris-serine precursor, e.g.9, followed by oxidation of the resulting tris-oxazoline intermediate, i.e.10. It is also plausible to speculate that the biosynthesis of metabolites related to ulapualide A, e.g. the bis oxazole 6 and the imide 7, involve cyclisations of just two of the serine units in 9. These speculations were given some credence by carrying out pertinent interconversions involving the bis-oxazole amide 24, the enamide 25, the imide 26, the oxazoline 27 and the tris-oxazole 30 as model compounds. An alternative strategy to the tris-oxazole macrolide intermediate 76 was also examined, involving preliminary synthesis of the aldehyde 73, containing a shortened (C25 C34) side chain from 67b and 47b. A Wadsworth-Emmons olefination reaction between 73 and the phosphonate ester 74 led smoothly to the E-alkene 75, but we were not able to reduce selectively the conjugated enone group in 75 to 76 without simultaneous reduction of the oxazole alkene bond, using a variety of reagents and reaction conditions. PMID- 18385856 TI - Stereoselective synthesis of (+)-polyoxamic acid based on the synthesis of chiral oxazine. AB - A concise, stereocontrolled synthesis of (+)-polyoxamic acid was achieved. Starting from trans-oxazoline as a chiral building block, the key step involves diastereoselective oxazine formation catalyzed by palladium(0). PMID- 18385857 TI - Enantioselective total synthesis of pyrinodemin A. AB - Pyrinodemin A 1, a cytotoxic marine alkaloid, was synthesized in a convergent and enantioselective fashion. The key steps are an asymmetric intramolecular dipolar cycloaddition of an oxazoline N-oxide to introduce the bicyclic ring system of the molecule, a cuprate coupling for the extension of the saturated chain and a B alkyl Suzuki coupling for the introduction of a 3-pyridyl moiety. Reductive amination allowed the coupling of the second side-chain onto the nitrogen atom to give 1. Additionally, attempts to prepare 1 from a trienic precursor by a double B-alkyl Suzuki reaction are described. PMID- 18385859 TI - Why green chemistry and sustainability of resources are essential to our future. PMID- 18385860 TI - From the 1996 groundwater action programme to the 2006 groundwater directive- what have we done, what have we learnt, what is the way ahead? AB - The need for action to avoid long term deterioration of fresh water quantity and quality was recognised by the Ministerial Seminar on groundwater held at The Hague in November 1991, which underlined the need to establish a programme of actions to be implemented by the year 2000 at national and Community level, aiming at sustainable management and protection of water resources. These discussions resulted in a communication from the European Commission in 1996 about a "Groundwater Action Programme" (GWAP). In parallel, a wide consultation had started on the need to streamline EU water policies, which led to the adoption of the Water Framework Directive in December 2000, later complemented by a "daughter" groundwater directive adopted in December 2006. This paper recalls the orientations of the GWAP and studies whether and how recommended actions have been tackled by the actual EU regulatory framework. PMID- 18385861 TI - 10th anniversary review: when healthy food becomes polluted-implications for public health and dietary advice. AB - Divergent scientific and regulatory agency perspectives about contaminants in fish have lead to contradictory advice and confusing public messages about its consumption. The evidence for the protective effect of eating fish on cardiovascular outcomes is considered to be convincing. It has long been attributed to n-3 unsaturated long-chain fatty acids. Persistent organic pollutants (POPs) are compounds that are lipid soluble and accumulate in the aquatic food chain. Despite a considerable decrease in their levels in fish during the last two decades, there is still significant concern about potential negative health effects and an ongoing debate exists about what type of fish consumption advisories are most suitable. In this review our aim is twofold, namely to explore: (1) the underlying causes for the conflicting recommendations by discussing the strengths and limitations of risk assessment and epidemiological evidence; and (2), the role of risk management in formulating public dietary advisories. It is our view that the latter advice is most appropriately formulated in the context of risk management, of which both epidemiologic evidence and risk assessment are essential components. PMID- 18385862 TI - Inorganic arsenic levels in rice milk exceed EU and US drinking water standards. AB - Under EU legislation, total arsenic levels in drinking water should not exceed 10 microg l(-1), while in the US this figure is set at 10 microg l(-1) inorganic arsenic. All rice milk samples analysed in a supermarket survey (n = 19) would fail the EU limit with up to 3 times this concentration recorded, while out of the subset that had arsenic species determined (n = 15), 80% had inorganic arsenic levels above 10 microg l(-1), with the remaining 3 samples approaching this value. It is a point for discussion whether rice milk is seen as a water substitute or as a food, there are no EU or US food standards highlighting the disparity between water and food regulations in this respect. PMID- 18385863 TI - Detection of polybrominated diphenyl ethers in tilapia (Oreochromis mossambicus) from O'ahu, Hawai;i. AB - Polybrominated diphenyl ethers (PBDEs) have been detected for the first time at a range from 231.58 to 685.61 ng g(-1) lipid weight in the muscles of tilapia (Oreochromis mossambicus) collected from O'ahu, an island of the geographically isolated Hawaiian archipelago. PMID- 18385864 TI - Solvation parameters for mercury and mercury(II) compounds: calculation of properties of environmental interest. AB - Descriptors have been determined for four inorganic mercury(II) species and for seventeen organic mercury(II) species, using experimental literature data. These descriptors can then be used in equations that we have already set out in order to estimate a large number of physicochemical properties. These include the water to octanol partition coefficient and the gas to water partition coefficient. For the organic mercury(II) species, including dimethylmercury and the methylmercury(II) halides, the latter has been estimated over the temperature range 273-373 K. PMID- 18385865 TI - Simplified determination of lipophilic metabolites of nonylphenol ethoxylates: method development and application in aqueous samples from Buenos Aires, Argentina. AB - In the present work we have developed an analytical methodology for the determination of nonylphenol (NP) and nonylphenol mono- and di-ethoxylates (NP1EO and NP2EO) in water samples. The applicability of this methodology was proved by means of the analysis of environmentally relevant aqueous samples from Buenos Aires. This constitutes a starting point for a rigorous assessment of the incidence of NPnEO surfactants in Argentina, as only very few, qualitative or semi-quantitative data on the occurrence of these compounds in local systems were available up to this time. Enrichment of the analytes was carried out by solid phase extraction on a C-18 sorbent, followed by elution with ethyl acetate. Normal-phase high performance liquid chromatography on an amino-silica column and fluorescence detection at excitation-emission wavelengths of 230-300 nm were employed for separation and quantification of the analytes. Confirmation of peak assignment in selected real samples was performed by off-line coupling HPLC with GC-MS analysis. A non-polar GC capillary column was used, and a characteristic peak pattern was obtained for the alkyl chain isomers of each ethoxylated homologue and NP. GC-MS analyses yielded in all cases purity levels higher than 80% for the HPLC collected fractions. The elevated concentrations found for the estrogenic metabolites of NPnEO are in accordance with an unrestricted use of this class of non-ionic surfactants in the country. PMID- 18385866 TI - Investigative monitoring within the European Water Framework Directive: a coastal blast furnace slag disposal, as an example. AB - The European Water Framework Directive (WFD) establishes a framework for the protection of estuarine and coastal waters, with the most important objective being to achieve 'good ecological status' for all waters, by 2015. Hence, Member States are establishing programmes for the monitoring of water quality status, through the assessment of ecological and chemical elements. These monitoring programmes can be of three types: surveillance monitoring; operational monitoring (both undertaken on a routine basis); and investigative monitoring (carried out where the reason of any exceedance for ecological and chemical status is unknown). Until now, nothing has been developed in relation to investigative monitoring and no clear guidance exists for this type of monitoring, as it must be tackled on a 'case-by-case' basis. Consequently, the present study uses slag disposal from a blast furnace, into a coastal area, as a case-study in the implementation of investigative monitoring, according to the WFD. In order to investigate the potential threat of such slags, this contribution includes: a geophysical study, to determine the extent of the disposal area; sediment analysis; a chemical metal analysis; and an ecotoxicological study (including a Microtox test and an amphipod bioassay). The results show that metal concentrations are several times above the background concentration. However, only one of the stations showed toxicity after acute toxicological tests, with the benthic communities being in a good status. The approaches used here show that contaminants are not bioavailable and that no management actions are required with the slags. PMID- 18385867 TI - Preliminary assessment of polybrominated diphenyl ethers (PBDEs) in the Scottish aquatic environment, including the Firth of Clyde. AB - This paper presents preliminary data on polybrominated diphenyl ethers (PBDEs) in the Scottish aquatic environment. Sediment and biota (fish liver, fish muscle and mussels) from a number of locations around Scotland were analysed for PBDEs with samples being from both remote and from potentially contaminated areas such as the former sewage sludge dump site at Garroch Head in the Clyde. PBDEs were measured in both cultivated, rope grown mussels and wild mussels collected from 5 sites around Scotland in 2006. Total PBDE concentrations (sum of tri- to hepta BDEs) ranged from 0.16 mg L(-1)), DTP (>0.24 mg L(-1)), NO3(-)-N (>0.15 mg L(-1)), NH4+-N (>0.12 mg L(-1)), and TKN (>0.96 mg L(-1)), occurred in the upstream of TMC, while high concentrations of chlorophyll a (>6.8 mug L(-l)) and pheophytin (>3.9 microg L(-l)) were detected in the downstream of TMC. The highest chlorophyll a (11.8 mug L(-l)) and pheophytin (6.06 microg L(-l)) concentrations, however, were associated with static and open water conditions. Hydrological parameters (total dissolved solid, electrical conductivity, salinity, pH, and water temperature) were positively correlated with chlorophyll a and pheophytin concentrations (P < 0.01) and these factors overshadowed the relationships between N and P concentrations and chlorophyll a under field conditions. Principal component analysis and the ratios of DIN/DP and TN/TP in the water suggest that N is the limiting nutrient factor for phytoplankton growth in the TMC and elevated N relative to P is beneficial to the growth of phytoplankton, which is supported by laboratory culture experiments under controlled conditions. PMID- 18385873 TI - Passive sampling of glycol ethers and their acetates in indoor air. AB - This study examined the performances of a thermal desorbable radial diffusive sampler for the weekly measurement of eight glycol ethers in indoor air and described the results of an application of this method carried out as part of HABIT'AIR Nord - Pas de Calais program for the air monitoring of these compounds in sixty homes located in northern France. The target compounds were the four glycol ethers banned from sale to the public in France since the 1990s (i.e. 2 methoxy ethanol, 2-ethoxy ethanol and their acetates) and four other glycol ethers derivatives of which the use have increased considerably (i.e. 1-methoxy-2 propanol, 2-butoxy ethanol and their acetates).A test program was carried out with the aim of validating the passive sampling method. It allowed the estimation of all the parameters of a method for each compound (calibration, analytical precision, desorption efficiency, sampling rate in standard conditions, detection limit and stability of sample before and after exposure), the examination of the influence of environmental factors on the sampling rate by some exposure chamber experiments and the assessment of the uncertainty of the measurements. The results of this evaluation demonstrated that the method has turned out to be suitable for six out of eight glycol ethers tested. The effect of the environmental factors on the sampling rates was the main source of measurement uncertainty. The measurements done in sixty homes revealed a relative abundance of 1-methoxy-2-propanol that was found in more than two thirds of homes at concentration levels of 4.5 microg m(-3) on average (a maximum value of 28 microg m(-3)). 1-methoxy-2-propanol acetate and 2-butoxy ethanol were also detected, but less frequently (in 19% of homes) and with the concentrations below 12 microg m( 3). The highest levels of these glycol ethers appear to be in relation to the emissions occurring at the time of cleaning tasks. PMID- 18385874 TI - Calibration of two passive air sampler configurations for monitoring concentrations of hexabromocyclododecanes in indoor air. AB - While polyurethane foam (PUF) disk passive air samplers are employed increasingly to monitor persistent organic pollutants in indoor air, they essentially sample only the vapour phase. As a previous investigation of the vapour : particle phase partitioning of hexabromocyclododecanes HBCDs in (outdoor) air reported them to be present largely in the particulate phase, we monitored three offices using active air samplers. In each, approximately 65% of HBCDs were present in the vapour phase, suggesting PUF disk passive samplers are suitable for monitoring HBCDs in indoor air. Concentrations in the three offices (239-359 pg Sigma HBCD m(-3)) exceed substantially those reported in outdoor air from the United States (2.1-11 pg Sigma HBCD m(-3)), but are in line with outdoor air from Stockholm. The relative abundance of the three principal diastereomers in office air was closer to that found in technical HBCD formulations (i.e. predominantly gamma HBCD) than in most US outdoor air samples. Time integrated air concentrations of alpha-, beta-, and gamma-HBCD were obtained for an office using a low volume sampler operated over a 50 d period alongside PUF disk samplers. This calibration exercise yielded the following passive air sampling rates for both a fully- and part-sheltered PUF disk sampler design: for alpha-, beta-, and gamma-HBCD, 0.87, 0.89, and 0.91 m3 d(-1) respectively (fully-sheltered) and 1.38, 1.54, and 1.55 m3 d(-1) respectively (part-sheltered). Deployment of the part-sheltered configuration yielded concentrations approximately 35% lower than those obtained using a high volume sampler, consistent with PUF disk samplers measuring primarily the vapour phase. PMID- 18385875 TI - Speciation/fractionation of nickel in airborne particulate matter: improvements in the Zatka sequential leaching procedure. AB - Modifications are reported to the sequential leaching analytical method for nickel speciation/fractionation specified by Zatka so that larger sample masses can be analyzed. Improvements have been made in the completeness of the sulfide/metallic separation during the peroxide-citrate leach step by use of a larger volume of leachant, a longer leach duration and an orbital shaker. Minimal extraction of metallic nickel in this prolonged sulfidic nickel extraction has been confirmed. An increase in the number of samples analyzed simultaneously using these modifications has resulted in substantial productivity improvements and concomitant lower costs. It is critical for practitioners of sequential leaching techniques to recognize potential limitations and to use professional judgment when interpreting results. For example, results obtained may not be biologically relevant in assessing health risks; the acts of sampling and storage may result in changes in fractionation with time; surface coatings/films may alter the ability of a leachant to react with the target compound; and leaching behaviours may be different for samples differing only in particle size distributions. PMID- 18385876 TI - Design and evaluation of an inlet conditioner to dry particles for real-time particle sizers. AB - Real-time particle sizers provide rapid information about atmospheric particles, particularly peak exposures, which may be important in the development of adverse health outcomes. However, these instruments are subject to erroneous readings in high-humidity environments when compared with measurements from filter-based, federal reference method (FRM) samplers. Laboratory tests were conducted to evaluate the ability of three inlet conditioners to dry aerosol prior to entering a real-time particle sizer for measuring coarse aerosols (Model 3321 Aerodynamic Particle Sizer, APS) under simulated highly humid conditions. Two 30 day field studies in Birmingham, AL, USA were conducted to compare the response of two APSs operated with and without an inlet conditioner to that measured with FRM samplers. In field studies, the correlation of PM(10-2.5) derived from the APS and that measured with the FRM was substantially stronger with an inlet conditioner applied (r2 ranged from 0.91 to 0.99) than with no conditioner (r2 = 0.61). Laboratory experiments confirmed the ability of the heater and desiccant conditioner to remove particle-borne moisture. In field tests, water was found associated with particles across the sizing range of the APS (0.5 microm to 20 microm) when relative humidity was high in Birmingham. Certain types of inlet conditioners may substantially improve the correlation between particulate mass concentration derived from real-time particle sizers and filter-based samplers in humid conditions. PMID- 18385877 TI - Chlorinated, brominated, and perfluorinated compounds, polycyclic aromatic hydrocarbons and trace elements in livers of sea otters from California, Washington, and Alaska (USA), and Kamchatka (Russia). AB - Concentrations of organochlorine pesticides (DDTs, HCHs, and chlordanes), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), perfluorinated compounds (PFCs), and 20 trace elements were determined in livers of 3- to 5-year old stranded sea otters collected from the coastal waters of California, Washington, and Alaska (USA) and from Kamchatka (Russia). Concentrations of organochlorine pesticides, PCBs, and PBDEs were high in sea otters collected from the California coast. Concentrations of DDTs were 10-fold higher in California sea otters than in otters from other locations; PCB concentrations were 5-fold higher, and PBDE concentrations were 2 fold higher, in California sea otters than in otters from other locations. Concentrations of PAHs were higher in sea otters from Prince William Sound than in sea otters from other locations. Concentrations of several trace elements were elevated in sea otters collected from California and Prince William Sound. Elevated concentrations of Mn and Zn in sea otters from California and Prince William Sound were indicative of oxidative stress-related injuries in these two populations. Concentrations of all of the target compounds, including trace elements, that were analyzed in sea otters from Kamchatka were lower than those found from the US coastal locations. PMID- 18385878 TI - Composition and concentration of hydrocarbons in sediment samples from the oil producing area of the East Shetland Basin, Scotland. AB - The East Shetland Basin is one of the areas that the Fisheries Research Services (FRS) has concentrated on to assess the possible impacts of oil exploration and production on the marine environment. A stratified random survey of the sediment was carried out in 2002. TOCs were low across the basin and were positively correlated with grain size. The total PAH concentrations (2- to 6-ring parent and alkylated PAHs, including the 16 US EPA PAHs) were less than 150 microg kg(-1) dry weight and their composition indicated a predominantly pyrolytic input to the basin in 2002. Minor unresolved complex mixtures in the n-alkane profiles indicated a slight petrogenic input but further examination of the biomarkers (hopanes and steranes) showed a mixed North Sea and Middle Eastern source. The Middle Eastern source is likely due to inputs from shipping activity, as it is widely used as bunker fuel. Grid surveys were carried out in 1986, 1988-89 and 1994 and areas were selected for which there was data for all the historic grid surveys and the 2002 stratified random survey. Although referring to only a small part of the East Shetland Basin, comparison with these historic surveys shows clearly that the concentrations of Forties crude oil equivalents and total PAH concentrations were highest in 1988-89 and by 2002 had returned to concentrations the same as or less than observed in the original survey in 1986. PMID- 18385879 TI - Monitoring the fall of large atmospheric ice conglomerations: a multianalytical approach to the study of the Mejorada del Campo megacryometeor. AB - Certain local atmospheric anomalies, such as the formation of unusually large ice conglomerations (megacryometeors), have been proposed to be a potential natural hazard for people and aviation, as well as geoindicators for fingerprinting larger-scale atmospheric environmental changes. On March 13th 2007, at approximately 10:15 am, an ice chunk weighing about 10 kg fell from the clear-sky and crashed through the roof (around 15 m) of an industrial storage house in Mejorada del Campo, a town located 20 km east from Madrid. The megacryometeor monitoring follow-up and the original investigation presented here includes, for the first time, both logistic and scientific collaboration between the Laboratory of the Environment, Criminalistic Service (SECRIM, the Spanish "Guardia Civil") and academic and scientific institutions (universities and the Spanish National Research Council). We propose that the management procedure of the incident, along with the detailed scientific research and combination of analytical methodologies in different laboratories, can serve as a protocol model for other similar events. PMID- 18385880 TI - The synthetic potential of pyridinium salt photochemistry. AB - The discovery in the 1970s by Kaplan, Wilzbach and Pavlik that pyridinium salts undergo a unique cyclization reaction to produce bicyclic-aziridines was virtually unrecognized for nearly three decades. It was only comparatively recently that the process was explored in more detail and its synthetic potential exploited. In this Perspective, photocyclization reactions of pyridinium salts will be discussed, starting with the initial discovery, covering related processes of pyrylium salts, and extending to applications to the synthesis of natural and non-natural products of biomedical interest. PMID- 18385881 TI - Water-soluble, pH-sensitive fluorescent probes on the basis of acridizinium ions. AB - The acridizinium fluorophore was employed as water-soluble substitute for anthracene in PET probes and donor-pi-acceptor systems that may be used as fluorescent light-up probes and ratiometric probes in aqueous media. PMID- 18385882 TI - Complexation of acridine orange by cucurbit[7]uril and beta-cyclodextrin: photophysical effects and pKa shifts. AB - The host-guest interactions of the neutral (AO) and cationic (AOH+) forms of the dye acridine orange with the macrocyclic hosts cucurbit[7]uril (CB7) and beta cyclodextrin (beta-CD) were investigated by using ground-state absorption and steady-state as well as time-resolved fluorescence measurements. The cationic form undergoes no significant complexation with beta-CD, but binds strongly with CB7 (Keq = 2.0 x 10(5) M(-1)), causing a large enhancement in fluorescence intensity and lifetime of the dye in the latter host. The strong and selective binding of AOH+ with CB7 is attributed to ion-dipole interactions involving the ureido carbonyl rims of CB7 and the charged AOH+. In contrast, the neutral AO form of the dye shows quite similar binding with both CB7 and beta-CD, but the binding constants are lower by more than two orders of magnitude compared to that of the AOH+-CB7 system. CB7 and beta-CD show a contrasting behavior in modifying the acid-base character of the dye, shifting its pKa by about 2.6 units upward and about 0.7 units downward, in the two respective cases. These divergent pKa shifts of the dye arise from the differential affinity of the two host molecules to encapsulate the protonated and neutral form of the dye. PMID- 18385883 TI - Sewage bacteriophage photoinactivation by cationic porphyrins: a study of charge effect. AB - Photodynamic therapy has been used to inactivate microorganisms through the use of targeted photosensitizers. Recently the inactivation of bacteria in residual waters has been reported, but nothing is known about photoinactivation of environmental bacteriophages, which are often used as indicators of human enteric viruses. In this study we tested the effect of six cationic porphyrin derivatives with two to four charges on the photoinactivation of a sewage bacteriophage. A phage suspension of 5 x 10(7) PFU mL(-1) was exposed to white light (40 W m(-2)), during 270 min, at three photosensitizer concentrations (0.5, 1.0 and 5.0 microM). Tetra- and tricationic porphyrins inactivated the T4-like sewage phage to the limits of detection, but dicationic porphyrins did not lead to a significant decrease in phage viability. At the highest photosensitizer concentration (5.0 microM), the phage was completely inactivated (>99.9999% of inactivation, reduction of 7.2 log) after 270 min by the tetracationic porphyrin. Two of the tricationic derivatives also led to phage inactivation to the limit of detection. The rate of bacteriophage photoinactivation and the efficiency of the photosensitizer appeared to vary with the charge and with the substituents in the meso-positions of the porphyrin macrocycle. Tetra- and tricationic porphyrins can, therefore, be used as a new method for inactivating sewage bacteriophages that are frequently used as human enteric virus indicators. The complete inactivation of viruses with low light intensity means that this methodology can be used even on cloudy days and during winter, opening the possibility to develop new technologies for wastewater treatment. PMID- 18385884 TI - An antenna-sensitised 1-acyl-7-nitroindoline that has good solubility properties in the presence of calcium ions and is suitable for use as a caged L-glutamate in neuroscience. AB - A new version of a benzophenone antenna-sensitised photolabile derivative of l glutamate, which has a dicarboxylic acid substituent on the benzophenone to promote water solubility, has been synthesised. It does not show problems of precipitation in the presence of calcium ions that were encountered with related compounds in which one or two phosphate groups were present as water-solubilising substituents but retains the enhanced photolytic efficiency that results from the benzophenone antenna. Photolysis of the compound proceeds with stoichiometric release of l-glutamate and pharmacological evaluations have shown that the compound itself has no evidence of agonist or antagonist activity in its unphotolysed form. PMID- 18385885 TI - Tissue distribution and pharmacokinetics of an ATWLPPR-conjugated chlorin-type photosensitizer targeting neuropilin-1 in glioma-bearing nude mice. AB - Destruction of the neovasculature is essential for efficient tumor eradication by photodynamic therapy (PDT). The PDT anti-vascular effect can be promoted by developing addressed photosensitizers localized preferentially to the tumor vascular compartment. A new photosensitizer conjugated to an heptapeptide [H-Ala Thr-Trp-Leu-Pro-Pro-Arg-OH (ATWLPPR)] targeting neuropilin-1, a Vascular Endothelial Growth Factor (VEGF) co-receptor, has been synthesized. It was administered intravenously for an easier access to endothelial cells lining the vasculature in human malignant glioma-bearing nude mice. Plasma pharmacokinetic parameters were derived from plasma concentration-time data using a non compartmental analysis and validated a relatively rapid elimination from the blood compartment with an elimination rate constant of 0.062 h(-1) and a biological half-life of 11.0 h. The photosensitizer was mainly concentrated in organs such as liver, spleen and kidneys, which are rich in reticuloendothelial cells. In these organs, the elimination profiles of the photosensitizer were comparable, with half-lives as short as 12.2, 15.1 and 19.7 h, respectively. The peptidic moiety of the conjugated photosensitizer was degraded to various rates depending on the organ considered, most of the degradation process occurred in organs of the reticuloendothelial system. A metabolic product resulting from the enzymatic cleavage of the peptide bond between Ala and Thr was detected in plasma at all the examined time points from 2 h post-injection. The conjugated photosensitizer accumulated rapidly and at high levels in the tumor, with 2.3% of injected dose per gram of tumor tissue at 1 h after injection. Taking into account the aspecific uptake of the degradation product, the tumor levels of total photoactivable compounds might exhibit an interesting photodynamic activity. On the contrary, levels of total photoactivable compounds remained low in the skin. This study provides essential information for the choice of the time interval not to exceed to activate the photosensitizer. PMID- 18385886 TI - Crystal structure of coelenterazine-binding protein from Renilla muelleri at 1.7 A: why it is not a calcium-regulated photoprotein. AB - Bioluminescence in the sea pansy Renilla involves two distinct proteins, a Ca2+ triggered coelenterazine-binding protein (CBP), and Renilla luciferase. CBP contains one tightly bound coelenterazine molecule, which becomes available for reaction with luciferase and O2 only subsequent to Ca2+ binding. CBP belongs to the EF-hand superfamily of Ca2+-binding proteins and contains three "EF-hand" Ca2+-binding sites. The overall spatial structure of recombinant selenomethionine labeled CBP determined at 1.7 A, is found to approximate the protein scaffold characteristic of the class of Ca2+-regulated photoproteins. Photoproteins however, catalyze molecular oxygen addition to coelenterazine producing a 2 hydroperoxycoelenterazine intermediate, which is stabilized within the binding cavity in the absence of Ca2+. Addition of Ca2+ triggers the bioluminescence reaction. However in CBP this first step of oxygen addition is not allowed. The different amino acid environments and hydrogen bond interactions within the binding cavity, are proposed to account for the different properties of the two classes of proteins. PMID- 18385887 TI - CCD imaging of basal bioluminescence in larval fireflies: clues on the anatomic origin and evolution of bioluminescence. AB - The anatomic and biochemical origin of beetle bioluminescence is still poorly understood. Through CCD imaging we report that larvae and pupae of the Brazilian fireflies Aspisoma lineatum and Cratomorphus sp emit a continuous weak glow throughout the entire body during all stages. This luminescence is especially developed after feeding, ecdysis and in the pupal stage, gradually disappearing as the cuticle becomes sclerotized and the adult emerges. This weak glow arises from the fat body, which consists of small lobes spread all over the body cavity. According to their pigmentation, these lobes can be divided in whitish and pinkish, and display different luciferase isozymes. Morphological studies suggest that the jelly-like ventral lanterns in the 8th abdominal segment evolved from these white lobes, providing a rationale for the widespread location of lanterns in larvae of different bioluminescent beetles. The biological and biochemical function of this weak diffuse bioluminescence is discussed in the context of the larval life-history. PMID- 18385888 TI - Kinetic analysis of the interaction between amphotericin B and human serum albumin using surface plasmon resonance and fluorescence spectroscopy. AB - The binding interaction between amphotericin B and human serum albumin (HSA) has been studied using surface plasmon resonance (SPR) spectroscopy combined with a fluorescence quenching method to confirm the binding kinetic results. In this paper, the SPR method used to study the drug-protein interaction has been described in detail. The association rate constant, dissociation rate constant and the equilibrium association constant of amphotericin B binding to HSA were obtained using this method. To confirm the feasibility of the SPR method, a fluorescence quenching method was performed to obtain the equilibrium constant. In order to obtain more accurate results, experiment design was used to optimize the fluorescence quenching process. The two equilibrium association constants obtained using the two methods were 4.017 x 10(4) M(-1) (SPR) and 3.656 x 10(4) M(-1) (fluorescence quenching method) respectively. PMID- 18385889 TI - Nano-patterned layers of a grafted coumarinic chromophore. AB - We report on the grafting of coumarin chromophores on flat silicon surfaces and in regions of nanometric dimensions drawn on silicon surfaces. The coumarin derivative was grafted by using the quaternization of a tertiary amine group of the chromophore with a ((chloromethyl)phenylethyl)-dimethylchlorosilane (CMPDCS) grafted on silicon. Complete characterization of the grafted layer was performed as a function of reaction time by X-ray photoelectron spectroscopy, X-ray reflectometry, atomic force microscopy, fluorescence spectroscopy and laser scanning confocal microscopy. The results indicate that about one chromophore molecule is grafted every second CMPDCS molecule, resulting in a surface density of coumarin of slightly more than one coumarin per nm2. A broadening of the distribution of the fluorescence lifetimes was observed, suggesting that the grafted molecules experience a larger distribution of environments in the grafted layer than in solution. Since this reaction is fully compatible with silicon processing technology, the grafting could also be performed in nano-regions of size as small as 250 nm defined by combining electron-beam lithography with silanization. In such nano-sized regions the distribution of fluorescence lifetimes was narrower, suggesting a possible influence of the confinement on the organization of the molecules. PMID- 18385890 TI - Laser flash photolysis study of the triplet reactivity of beta-lapachones. AB - The photochemical reactivity of beta-lapachone (1), nor-beta-lapachone (2) and beta-lapachone 3-sulfonic acid (3) has been examined by laser flash photolysis. Excitation (lambda = 266 nm) of degassed solutions of , in acetonitrile or dichloromethane, resulted in the formation of detectable transients with absorption maxima at 300, 380 and 650 nm. These transients, with lifetimes of 5.0 micros, were quenched by beta-carotene at a diffusion-controlled rate constant and assigned to the triplet excited states of 1-3. Addition of hydrogen donors, such as 2-propanol, 1,4-cyclohexadiene, 4-methoxyphenol or indole led to the formation of new transients, which were assigned to the corresponding ketyl radicals obtained from the hydrogen abstraction reaction by the triplets 1-3 . In the presence of triethylamine it was observed the formation of the long-lived anion radical derived from , which shows absorption maxima at 300 and 380 nm. The low values observed for the hydrogen abstraction rate constants for the beta lapachones 1-3 using 2-propanol and 1,4-cyclohexadiene as quenchers led us to conclude that their triplet excited states show pi pi* character. PMID- 18385891 TI - Near-infrared luminescence properties of erbium complexes with the substituted phthalocyaninato ligands. AB - Two homoleptic bis(phthalocyaninato) erbium(III) complexes Er[Pc(beta-OR/R)4]2 and two half-sandwich phthalocyaninato erbium(III) complexes (acac)Er[Pc(beta OR/R)4] (OR = 1-n-pentyloxy and R = tert-butyl) have been investigated. Then we studied the near-infrared luminescence properties of the compounds. When the phthalocyanine ligands were excited, half-sandwich phthalocyaninato erbium(III) complexes showed strong near-infrared luminescence at 1540 nm while homoleptic bis(phthalocyaninato) erbium(III) complexes showed no signals. The full width at half maximum (FWHM) centered at 1540 nm in the emission spectrum of the half sandwich complexes in solution and in solid state are 67 nm and 78 nm respectively, which shows potential for optical-amplification applications. Erbium-doped standard polymethyl-methacrylate (PMMA) matrix which was synthesized has also displayed NIR emission. PMID- 18385892 TI - Solvent effect on the physical quenching of singlet molecular oxygen by p quinones. AB - Rate constants for the interaction between singlet molecular oxygen [O2(1 delta g)] and the p-quinones 1,4-benzoquinone (BQ), duroquinone (DQ), 9,10 anthraquinone (AQ) and 1,8-dihydroxy-9,10-anthraquinone (OHAQ) are reported for several solvents at room temperature. The solvent effect on the total quenching rate constant (kt) was analysed employing the semiempirical solvatochromic equation proposed by Kamlet and Taft. The higher values of kt (2-7 x 10(7) M(-1) s(-1)) were obtained when the hydrogen-bond donor solvent ability is increased (higher alpha parameter values). The results indicate the importance of specific solvent interactions in governing the rates of the quenching. PMID- 18385893 TI - Heat-induced changes in photosystem I activity as measured with different electron donors in isolated spinach thylakoid membranes. AB - Heat-induced changes in photosystem I (PSI) have been studied in terms of rates of oxygen consumption using various donors (DCPIPH2, TMPDred and DADred), formation of photo-oxidized P700 and changes in Chl a fluorescence emission at 77 K. Linear heating of thylakoid membranes from 35 degrees C to 70 degrees C caused an enhancement in PSI-mediated electron transfer rates (DCPIPH2-->MV) up to 55 degrees C. However, no change was observed in PSI rates when other electron donors were used (TMPDred and DADred). Similarly, Chl a fluorescence emission spectra at 77 K of heat-treated thylakoid membranes did not show any increase in peak at 735 nm, however, a significant decrease was observed as a function of temperature in the peaks at 685 and 694 nm. In DCMU-treated control thylakoid membranes maximum photo-oxidized P700 was generated at g = 2.0025. In heat treated thylakoid membranes maximum intensity of photo-oxidized P700 signal was observed at approximately 50-55 degrees C without DCMU treatment. The steady state signal of the photo-oxidized P700 was studied in the presence of DCPIPH2 and TMPDred as electron donors in DCMU-treated control and in 50 degrees C treated thylakoid membranes. We present here the first of such comparative study of PSI activity in terms of the rates of oxygen consumption and re-reduction kinetics of photo-oxidized P700 in the presence of different electron donors. It appears that the formation of the P700+ signal in heat-treated thylakoid membranes is due to an inhibited electron supply from PSII and not due to spillover or antenna migration. PMID- 18385894 TI - Identification of 3,4-didehydrorhodopin as major carotenoid in Rhodopseudomonas species. AB - Recently we isolated the purple photosynthetic bacterium, Rhodopseudomonas sp. Rits, which was phylogenetically related to Rhodopseudomonas (Rps.) palustris. In this study, the light-dependent and time-dependent changes in the carotenoid composition were investigated by HPLC analysis of extracts from the cultures. All seven carotenoids in the biosynthetic pathway from lycopene to spirilloxanthin were detected. Especially, 3,4-didehydrorhodopin, having twelve conjugated double bonds as well as one terminal hydroxy group, was isolated in a remarkably large amount and fully characterized for the first time. The biosynthetic intermediate was commonly found in the Rps. palustris strains (CGA009, Morita and NBRC100419). PMID- 18385895 TI - Chlorophyll fluorescence emission spectrum inside a leaf. AB - Chlorophyll a fluorescence can be used as an early stress indicator. Fluorescence is also connected to photosynthesis so it can be proposed for global monitoring of vegetation status from a satellite platform. Nevertheless, the correct interpretation of fluorescence requires accurate physical models. The spectral shape of the leaf fluorescence free of any re-absorption effect plays a key role in the models and is difficult to measure. We present a vegetation fluorescence emission spectrum free of re-absorption based on a combination of measurements and modelling. The suggested spectrum takes into account the photosystem I and II spectra and their relative contribution to fluorescence. This emission spectrum is applicable to describe vegetation fluorescence in biospectroscopy and remote sensing. PMID- 18385896 TI - Hormones and sport: physiology, pharmacology and forensic science. PMID- 18385897 TI - The physiological and pharmacological basis for the ergogenic effects of androgens in elite sports. AB - Androgen doping in power sports is undeniably rampant worldwide. There is strong evidence that androgen administration in men increases skeletal muscle mass, maximal voluntary strength and muscle power. However, we do not have good experimental evidence to support the presumption that androgen administration improves physical function or athletic performance. Androgens do not increase specific force or whole body endurance measures. The anabolic effects of testosterone on the skeletal muscle are mediated through androgen receptor signaling. Testosterone promotes myogenic differentiation of multipotent mesenchymal stem cells and inhibits their differentiation into the adipogenic lineage. Testosterone binding to androgen receptor induces a conformational change in androgen receptor protein, causing it to associate with beta-catenin and TCF-4 and activate downstream Wnt target genes thus promoting myogenic differentiation. The adverse effects of androgens among athletes and recreational bodybuilders are under reported and include acne, deleterious changes in the cardiovascular risk factors, including a marked decrease in plasma high-density lipoproteins (HDL) cholesterol level, suppression of spermatogenesis resulting in infertility, increase in liver enzymes, hepatic neoplasms, mood and behavioral disturbances, and long term suppression of the endogenous hypothalamic-pituitary gonadal axis. Androgens are often used in combination with other drugs which may have serious adverse events of their own. In spite of effective methods for detecting androgen doping, the policies for screening of athletes are highly variable in different countries and organizations and even existing policies are not uniformly enforced. PMID- 18385898 TI - Effects of testosterone replacement and its pharmacogenetics on physical performance and metabolism. AB - In men, testosterone (T) deficiency is associated with decreased physical performance, as defined by adverse traits in body composition, namely increased body fat content and reduced muscle mass. Physical abilities in androgen deficient men are further attenuated by lower oxygen supply due to decreased hemoglobin concentrations and by poor glucose utilization. Dysthymia and a lack of necessary aggressiveness also contribute to deteriorate physical effectiveness. Substitution of T can improve lipid and insulin metabolism as well as growth of muscle fibers and decreasing fat depots, which consequently will result in changes of body composition. Increment of bone density will further contribute to increase physical fitness. The effects of T replacement therapy (TRT) are strongly influenced by age, training, and also pharmacogenetics: the CAG repeat polymorphism in exon 1 of the androgen receptor (AR) gene modulates androgen effects. In vitro, transcription of androgen-dependent target genes is attenuated with increasing length of triplet residues. Clinically, the CAG repeat polymorphism causes significant modulations of androgenicity in healthy eugonadal men as well as efficacy of TRT. Thresholds at which T treatment should be initiated, as well as androgen dosage, could be tailored according to this polymorphism. PMID- 18385900 TI - Androgenic-anabolic steroids and the Olympic Games. AB - Androgenic-anabolic steroids (AAS) have been misused by athletes at the Olympic Games, both before and after they were prohibited in sport in 1974. Systematic doping with AAS occurred in the German Democratic Republic (GDR) from 1965 to 1989 which assisted that country to win many medals at Olympic Games, especially in female events. Currently, AAS are the most frequent category of prohibited substances detected in the urine of athletes both globally and at the last two Summer Olympic Games. Scientific confirmation that AAS are effective in enhancing sports performance was difficult because ethical approval was difficult for research involving male subjects taking massive doses of androgens as some athletes and bodybuilders did. Methods to detect AAS have evolved gradually over the past three decades and currently, despite an impressive array of sophisticated analytical equipment and methods, anti-doping authorities and analytical scientists continue to face challenges as have occurred from the use by athletes of designer AAS during the past few years. The future development and use of selective androgen receptor modulators (SARMs) can be anticipated to pose problems in the years ahead. Endocrinologists should be aware that on occasions, replacement testosterone (T) therapy may be authorized in sport as a therapeutic use exemption (TUE) and these circumstances are discussed. PMID- 18385899 TI - Physiological and pharmacological basis for the ergogenic effects of growth hormone in elite sports. AB - Growth Hormone (GH) is an important and powerful metabolic hormone that is secreted in a pulsatile pattern from cells in the anterior pituitary, influenced by several normal and pathophysiological conditions. Human GH was first isolated in the 1950s and human derived cadaveric GH was initially used to treat patients with GH deficiency. However, synthetic recombinant GH has been widely available since the mid-1980s and the advent of this recombinant GH boosted the abuse of GH as a doping agent. Doping with GH is a well-known problem among elite athletes and among people training at gyms, but is forbidden for both medical and ethical reasons. It is mainly the anabolic and, to some extent, the lipolytic effects of GH that is valued by its users. Even though GH's rumour as an effective ergogenic drug among athletes, the effectiveness of GH as a single doping agent has been questioned during the last few years. There is a lack of scientific evidence that GH in supraphysiological doses has additional effects on muscle exercise performance other than those obtained from optimised training and diet itself. However, there might be synergistic effects if GH is combined with, for example, anabolic steroids, and GH seems to have positive effect on collagen synthesis. Regardless of whether or not GH doping is effective, there is a need for a reliable test method to detect GH doping. Several issues have made the development of a method for detecting GH doping complicated but a method has been presented and used in the Olympics in Athens and Turin. A problem with the method used, is the short time span (24-36 hours) from the last GH administration during which the test effectively can reveal doping. Therefore, out-of-competition testing will be crucial.However, work with different approaches to develop an alternative, reliable test is ongoing. PMID- 18385901 TI - Hormone abuse in sports: the antidoping perspective. AB - Since ancient times, unethical athletes have attempted to gain an unfair competitive advantage through the use of doping substances. A list of doping substances and methods banned in sports is published yearly by the World Anti Doping Agency (WADA). A substance or method might be included in the List if it fulfills at least two of the following criteria: enhances sports performance; represents a risk to the athlete's health; or violates the spirit of sports. This list, constantly updated to reflect new developments in the pharmaceutical industry as well as doping trends, enumerates the drug types and methods prohibited in and out of competition. Among the substances included are steroidal and peptide hormones and their modulators, stimulants, glucocorticosteroids, beta2-agonists, diuretics and masking agents, narcotics, and cannabinoids. Blood doping, tampering, infusions, and gene doping are examples of prohibited methods indicated on the List. From all these, hormones constitute by far the highest number of adverse analytical findings reported by antidoping laboratories. Although to date most are due to anabolic steroids, the advent of molecular biology techniques has made recombinant peptide hormones readily available. These substances are gradually changing the landscape of doping trends. Peptide hormones like erythropoietin (EPO), human growth hormone (hGH), insulin, and insulin-like growth factor I (IGF-I) are presumed to be widely abused for performance enhancement. Furthermore, as there is a paucity of techniques suitable for their detection, peptide hormones are all the more attractive to dishonest athletes. This article will overview the use of hormones as doping substances in sports, focusing mainly on peptide hormones as they represent a pressing challenge to the current fight against doping. Hormones and hormones modulators being developed by the pharmaceutical industry, which could emerge as new doping substances, are also discussed. PMID- 18385902 TI - Androgen abuse in sports. AB - Androgens remain the most effective and widely abused ergogenic drugs in sport. Although androgen doping has been prohibited for over 3 decades with a ban enforced by mass spectrometric (MS)-based urine testing for synthetic and exogenous natural androgens, attempts continue to develop increasingly complex schemes to circumvent the ban. A prominent recent approach has been the development of designer androgens. Such never-marketed androgens evade detection because mass spectrometry relies on identifying characteristic chemical signatures requiring prior knowledge of chemical structure. Although once known, designer androgens are readily detected and added to the Prohibited List. However, until their structures are elucidated, designer androgens can circumvent the ban on androgen doping. To combat this, in vitro androgen bioassays offer powerful new possibilities for the generic detection of unidentified bioactive androgens, regardless of their chemical structure. Another approach to circumvent the ban on androgen doping has been the development of indirect androgen doping, the use of exogenous drugs to produce a sustained increase in endogenous testosterone (T) production. Apart from estrogen blockers, however, such neuroendocrine active drugs mostly provide only transient increases in blood T. Finally the ban on androgen doping must allow provision for rare athletes with incidental, proven androgen deficiency who require T replacement therapy. The Therapeutic Use Exemption mechanism makes provision for such necessary medical treatment, subject to rigorous criteria for demonstrating a genuine ongoing need for T and monitoring of T dosage. Effective deterrence of sports doping requires novel, increasingly sophisticated detection options calibrated to defeat these challenges, without which fairness in sport is tarnished and the social and health idealization of sporting champions devalued. PMID- 18385903 TI - A robust test for growth hormone doping--present status and future prospects. AB - Although doping with growth hormone (GH) is banned, there is anecdotal evidence that it is widely abused. GH is reportedly used often in combination with anabolic steroids at high doses for several months. Development of a robust test for GH has been challenging because recombinant human 22 kDa (22K) GH used in doping is indistinguishable analytically from endogenous GH and there are wide physiological fluctuations in circulating GH concentrations. One approach to GH testing is based on measurement of different circulating GH isoforms using immunoassays that differentiate between 22K and other GH isoforms. Administration of 22K GH results in a change in its abundance relative to other endogenous pituitary GH isoforms. The differential isoform method has been implemented; however, its utility is limited because of the short window of opportunity of detection. The second approach, which will extend the window of opportunity of detection, is based on the detection of increased levels of circulating GH responsive proteins, such as insulin-like growth factor (IGF) axis and collagen peptides. Age and gender are the major determinants of variability for IGF-I and the collagen markers; therefore, a test based on these markers must take age into account for men and women. Extensive data is now available that validates the GH responsive marker approach and implementation is now largely dependent on establishing an assured supply of standardized assays. Future directions will include more widespread implementation of both approaches by the World Anti Doping Agency, possible use of other platforms for measurement and an athlete's passport to establish individual reference levels for biological parameters such as GH-responsive markers. Novel approaches include gene expression and proteomic profiling. PMID- 18385904 TI - Olympic sports and transsexuals. AB - Sex segregation in competitive sports is regarded as fair. Before puberty boys and girls do not differ in height, muscle and bone mass. Testosterone (T) exposure during puberty leads to an ultimate average greater height in men of 12 15 cm, longer and larger bones and muscle mass and strength and higher hemoglobin levels. Postpubertal androgen ablation reverses, at least in part, previous anabolic effects of T on muscle, bone mineral density and hemoglobin but the long bones remain longer and wider. T administration dose dependently increases muscle mass and maximal voluntary strength. Therefore, exogenous androgens, being performance enhancing drugs, are banned for all athletes. An issue is the participation in competitive sports of people with errors of sexual differentiation and particularly transsexuals who have been sex-reassigned. In view of the effects of T, a clear demarcation is whether sex reassignment has taken place before or after hormonal puberty. Pubertal effects of T are in part reversible but there is no reliable evidence as to its completeness. The International Olympic Committee (IOC) has taken an inevitably arbitrary decision with regard to participation of sex-reassigned transsexuals in elite sports: sex reassignment must have taken place at least two years earlier, hormone treatment must be appropriate for the reassigned sex and the reassigned sex must be legally recognized. The IOC policy is not binding for other organizations. PMID- 18385905 TI - Molecular Yin and Yang of erectile function and dysfunction. AB - In regard to erectile function, Yin is flaccidity and Yang erection. In the past decade, research has mostly focused on the Yang aspect of erectile function. However, in recent years, the Yin side is attracting increasingly greater attention. This is due to the realization that penile flaccidity is no less important than penile erection and is actively maintained by mechanisms that play critical roles in certain types of erectile dysfunction (ED); for example, in diabetic patients. In addition, there is evidence that the Yin and Yang signaling pathways interact with each other during the transition from flaccidity to erection, and vice versa. As such, it is important that we view erectile function from not only the Yang but also the Yin side. The purpose of this article is to review recent advances in the understanding of the molecular mechanisms that regulate the Yin and Yang of the penis. Emphasis is given to the Rho kinase signaling pathway that regulates the Yin, and to the cyclic nucleotide signaling pathway that regulates the Yang. Discussion is organized in such a way so as to follow the signaling cascade, that is, beginning with the extracellular signaling molecules (e.g., norepinephrin and nitric oxide) and their receptors, converging onto the intracellular effectors (e.g., Rho kinase and protein kinase G), branching into secondary effectors, and finishing with contractile molecules and phosphodiesterases. Interactions between the Yin and Yang signaling pathways are discussed as well. PMID- 18385906 TI - Clinical application of a new device for minimally invasive circumcision. AB - AIM: To study the clinical effects of a disposable circumcision device in treatment of male patients of different ages with either phimosis or excess foreskin. METHODS: One thousand two hundred patients between the age of 5 and 95 years underwent circumcision using this procedure in the 2-year period between October 2005 and September 2007. Of these cases, 904 had excess foreskin and 296 were cases of phimosis. RESULTS: In 96.33% of the cases the incision healed, leaving a minimal amount of the inner foreskin with no scarring and producing good cosmetic results. There were no incidents of device dislocation or damage to the frenulum. The average operative time was 2.5 min for excess foreskin, and 3.5 min for phimosis. During the 7 days of wearing the device, mild to moderate edema occurred in 10.08% of cases with excess foreskin and in 2.58% of those with phimosis. Edema in the frenulum was seen in 1.67% of patients, and only 0.67% had an infection of the incision. A total of 86.25% of patients reported pain due to penile erection. After removal of the device, 0.58% of the cases had minimal bleeding around the incision, and 2.42% had wound dehiscence. CONCLUSION: The new device can be applied to an overwhelming majority of patients with phimosis and excess foreskin. This technique is relatively simple to perform, and patients who underwent this surgery had very few complications. Antibiotics were not required and patients reported less pain than those who were circumcised using conventional methods. Circumcision with this device requires minimal tissue manipulation, and is quicker and safer than circumcision using conventional techniques. PMID- 18385907 TI - Stability of fluorochrome based assays to measure subcellular sperm functions. AB - AIM: To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. METHODS: Semen samples from 87 unselected infertile patients were used to perform the following assays: (i) detection of active caspase-3 (n=17); (ii) integrity of the mitochondrial membrane potential (MMP) (n=17); (iii) externalization of phosphatidylserine (EPS) (n=16); and (iv) detection of intact acrosomes via CD46 (n=37). After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14. RESULTS: Differences of up to +/-5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA showed mean differences<5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences>5% at day 3. The CD46-FITC labeling displayed absolute differences<5% CD46-positive spermatozoa at days 3, 7, 10 and 14. CONCLUSION: Although immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day. PMID- 18385908 TI - Novel UBE2B-associated polymorphisms in an azoospermic/oligozoospermic population. AB - AIM: To assess whether abnormalities exist in the UBE2B gene in a population of infertile human males, and to establish biologic plausibility of any discovered mutations. METHODS: We carried out polymerase chain reaction (PCR) amplification and sequence analysis of the 5'-untranslated region and six exons of the UBE2B gene, including flanking intronic regions, in a group of fertile and infertile men. Following the identification of a putative promoter region that contained single or dual triplet deletions within a 10-CGG repeat island, we evaluated the binding affinity of these identified polymorphisms as compared to the wild-type sequence to transcription factor SP1 using a DNA-protein gel shift assay. RESULTS: There was a novel exonic single nucleotide polymorphism (SNP) noted in exon 4 in 5% of infertile men. In silico 3D modeling of the altered protein showed an innocuous isoleucine for valine substitution. There were no mutations noted within any of the other exons. Three novel intronic SNPs were identified within the fertile group, and seven novel intronic SNPs identified in the infertile group. The DNA-protein gel shift assay noted that both single CGG deletion and double CGG deletion bands had approximately twice the binding affinity compared to the wild-type for SP1. The negative control confirmed no non specific protein binding. CONCLUSION: By themselves, a single or double CGG deletion is unlikely to pose biologic significance. However, such deletions in this suspected promoter region are associated with increased binding affinity for SP1, and might represent one of several factors required for alteration of UBE2B gene expression. PMID- 18385909 TI - Detection of TMPRSS2:ERG fusion gene in circulating prostate cancer cells. AB - AIM: To investigate the existence of TMPRSS2:ERG fusion gene in circulating tumor cells (CTC) from prostate cancer patients and its potential in monitoring tumor metastasis. METHODS: We analyzed the frequency of TMPRSS2:ERG and TMPRSS2:ETV1 transcripts in 27 prostate cancer biopsies from prostatectomies, and TMPRSS2:ERG transcripts in CTC isolated from 15 patients with advanced androgen independent disease using reverse transcription polymerase chain reaction (RT-PCR). Fluorescence in situ hybridization (FISH) was applied to analyze the genomic truncation of ERG, which is the result of TMPRSS2:ERG fusion in 10 of the 15 CTC samples. RESULTS: TMPRSS2:ERG transcripts were found in 44% of our samples, but we did not detect expression of TMPRSS2:ETV1. Using FISH analysis we detected chromosomal rearrangements affecting the ERG gene in 6 of 10 CTC samples, including 1 case with associated TMPRSS2:ERG fusion at the primary site. However, TMPRSS2:ERG transcripts were not detected in any of the 15 CTC samples, including the 10 cases analyzed by FISH. CONCLUSION: Although further study is required to address the association between TMPRSS2:ERG fusion and prostate cancer metastasis, detection of genomic truncation of the ERG gene by FISH analysis could be useful for monitoring the appearance of CTC and the potential for prostate cancer metastasis. PMID- 18385910 TI - Effect of 43 degrees treatment on expression of heat shock proteins 105, 70 and 60 in cultured monkey Sertoli cells. AB - AIM: To examine the possible effect of heat treatment on expression of heat shock proteins (Hsps) 105, 70, and 60 in primary monkey Sertoli cells and to evaluate the possible signal pathways. METHODS: Western blot analysis, real-time polymerase chain reaction (PCR), and confocal immunohistochemistry were used to analyze mRNA and protein levels of the Hsps in response to 43 degrees treatment of Sertoli cells isolated from pubertal monkey testes. RESULTS: Staining with Hoechst 33342 indicated Sertoli cells did not undergo apoptosis after heat treatment. Hsp105 was expressed in cytoplasm of untreated Sertoli cells. Both Hsp105 mRNA and protein levels were increased approximately 20-fold compared to those of the untreated controls at 12 h after heat treatment. Untreated Sertoli cells did not express Hsp70, but heat stress induced its expression in the cell cytoplasm. The time-course of changes in Hsp70 was similar to that of Hsp105. In contrast to Hsp105 and Hsp70, the change in Hsp60 expression was much less obvious. The protein level between 12 h and 48 h after heat treatment was only approximately 1.5-fold that of the untreated control. Extracellular regulated kinase (ERK) 1/2 inhibitor (U0126) or phosphoinositide kinase-3 (PI3K) inhibitor (LY294002) could partially block the response of Hsp105 and Hsp70 induced by heat treatment. CONCLUSION: These results indicate that the heat-induced expression of the three types of Hsp in monkey Sertoli cells might be regulated by ERK and/or PI3K signal pathways, but the profile of their expression is different, suggesting that they might have different regulatory functions in Sertoli cells. PMID- 18385911 TI - Effect of vasectomy via inguinal canal on spermatogenesis in rabbits. AB - AIM: To determine whether vasectomy away from the epididymal tail (via the inguinal canal) in rabbits can reduce the early postoperative effects on spermatogenesis. METHODS: Twenty-nine normal male Japanese white rabbits (aged 4 6 months) were subjected to unilateral close-ended (conventional) or open-ended (the cut end of the juxta-epididymal vas deferens not ligated) vasectomy via the inguinal canal. Ten days and 3 months after operation, testes, epididymides and vasa deferentia were removed and methacrylate resin-embedded sections prepared. The histology of the testis, epididymis and vas deferens was examined under light microscope, and the volume and diameter of the seminiferous tubules were quantitatively studied using stereological methods. RESULTS: Neither of the methods of vasectomy led to apparent damage to spermatogenesis on the vasectomized side in comparison with the contralateral sham-operated side, but the juxta-epididymal vas deferens on the vasectomized side was highly distended and contained numerous sperm 3 months after operation. CONCLUSION: Vasectomy away from the cauda epididymis has no significant early postoperative effects on spermatogenesis in rabbits. PMID- 18385912 TI - Efficacy and tolerability of vardenafil in Asian men with erectile dysfunction. AB - AIM: To evaluate the efficacy and tolerability of vardenafil, a phosphodiesterase type-5 (PDE-5) inhibitor, in men of Asian ethnicity with erectile dysfunction (ED). METHODS: In this prospective, double-blind, multinational study, Asian men were randomized to receive vardenafil (10 mg) or placebo (4:1 ratio) for 12 weeks. The primary efficacy variables were the International Index of Erectile Function erectile function domain (IIEF-EF), and Sexual Encounter Profile (SEP) questions related to penetration and intercourse completion. Significant mean improvements were required in all three measures to show positive benefits of vardenafil treatment. Secondary efficacy variables included the Global Assessment Question (GAQ) on erection improvement. RESULTS: Least-squares mean baseline IIEF EF domain scores (vardenafil 14.6, placebo 13.4) were consistent with moderate ED. After 12 weeks, vardenafil treatment was associated with significant increases from the baseline in IIEF-EF domain scores compared with the placebo (22.4 vs. 14.3; P<0.001). Vardenafil was associated with significant improvements from baseline in least squares (LS) mean success rates for SEP-2 (vardenafil 82.2 vs. placebo 43.6; P<0.001) and SEP-3 (vardenafil 66.1 vs. placebo 24.0; P<0.001). Positive GAQ responses were reported by 81.8% of vardenafil recipients vs. 24.3% of placebo recipients. Adverse events were reported by 25.4% of the vardenafil group, the majority mild and transient. CONCLUSION: Vardenafil (10 mg) is a highly effective and well-tolerated treatment for moderate ED in Asian men. These results add to the increasing amount of data demonstrating the safety and efficacy of vardenafil for the treatment of ED in a range of patient populations. PMID- 18385913 TI - Comparison of tiotropium bromide and combined ipratropium/salbutamol for the treatment of COPD: a UK General Practice Research Database 12-month follow-up study. AB - AIMS: To compare the effectiveness of the long-acting anticholinergic, tiotropium with ipratropium/salbutamol in reducing the risk of exacerbations and COPD related referrals in patients with COPD. METHODS: Data were obtained from the General Practice Research Database (GPRD). Propensity score matching was used to balance prognostic covariates between treatment groups. Incidence rate ratios and 95% confidence intervals during a 12-month follow-up period were estimated. RESULTS: 4193 patients (3385, tiotropium; 808, ipratropium/salbutamol) in the GPRD met the inclusion/exclusion criteria. Patients treated with tiotropium had more severe COPD than patients treated with ipratropium/salbutamol. Following propensity score matching, 1222 tiotropium-treated patients and 633 ipratropium/salbutamol-treated patients were included in the final analysis. Incidence rate ratios (95% confidence intervals) were 0.74 (0.64-0.85; p=0.0086) for exacerbations and 0.57 (0.46-0.70; p=0.004) for COPD-related referrals/hospitalisations. CONCLUSIONS: Tiotropium is associated with a reduced risk of exacerbations and COPD-related referrals and hospitalisation compared to combined ipratropium/salbutamol in patients with COPD. PMID- 18385914 TI - [Fistulas and fissures. Part I: perianal fistulas]. AB - Perianal abscesses are caused by cryptoglandular infections at the dentate line between the anal sphincters. Acute therapy will relieve the pain but not the development of perianal fistulas. The challenge in therapy of perianal fistulas balances between the best possible cure and the preservation of continence. Local treatment with fibrin glue is a first step whenever continence might be endangered by operative procedures. First results with fistula "plugs" are promising but need further critical observation. Lower, intersphincteric fistulas can be treated by fistulotomy without risking a substantial loss in continence, but higher, suprasphincteric or complex fistula systems might be treated as a first step with a seton--followed by surgery as a second step. Excision of the external fistula tract, closure of the internal opening, and a local advancement flap are now competing with fistulotomy, curettage, and immediate reconstruction. PMID- 18385915 TI - [Lupus vulgaris manifestation as a destructive nose and facial tumor]. AB - Lupus vulgaris is the most frequent manifestation of cutaneous tuberculosis, but in Europe it is limited to isolated cases. Mainly immunocompetent individuals are affected by this result of an endogenous reinfection on a lymphogenous-less frequently hematogenous-pathway. Lupus vulgaris has been observed to develop in more than 50% of all patients who already suffer from other manifestations of tuberculosis. The development of a squamous cell carcinoma in the lupus vulgaris is a rare complication; therefore, lupus vulgaris is deemed a facultative precancerosis.A 68-year-old female Serbo-Croatian patient presented with an extensive ulcerative nose and facial tumor. Her anamnesis included a squamous cell carcinoma of the nose that had been excised alio loco 3 years before. Further examinations revealed enlarged cervical lymphoma on both sides, and pulmonary metastases were also suspected. The tumor biopsy revealed a necrotic, granulomatous inflammation. No acid-fast rods were seen on Ziehl-Neelsen stain. The tuberculous origin of this ulcerative skin tumor-the lupus vulgaris-as an endogenous reinfection of pulmonary tuberculosis manifestation was confirmed by the detection of Mycobacterium tuberculosis DNA in polymerase chain reaction and the growth of Mycobacterium tuberculosis colonies in the bacterial culture (skin biopsy and bronchial secretion). The skin tumor as well as the pulmonary manifestation were successfully treated with combined tuberculostatic therapy and showed a dramatic response within 3 months. PMID- 18385916 TI - Estrogen receptor alpha polymorphism is associated with pelvic organ prolapse risk. AB - Estrogen and estrogen receptors are known to play important roles in the pathophysiology of pelvic organ prolapse (POP). We investigated whether estrogen receptor alpha (ERalpha) gene polymorphisms were associated with POP risk by conducting a case-control association study in 88 women with POP and 153 women without POP. Genotypes of the ERalpha (ESR1) gene polymorphisms (rs17847075, rs2207647, rs2234693, rs3798577, and rs2228480) were determined by polymerase chain reaction, followed by restriction fragment length polymorphism analysis. There was significant difference between women with and those without POP in the distribution of the ESR1 rs2228480 genotypes evaluated. By using multivariable logistic regression, age and ESR1 rs2228480 genotype GA were significantly associated with POP risk. Although the sample size of women with POP studied is small, the present study shows that ERalpha genotype may be associated with POP risk. PMID- 18385917 TI - Primary repair of advanced obstetric anal sphincter tears: should it be performed by the overlapping sphincteroplasty technique? AB - Advanced obstetric anal sphincter tears are often associated with a high incidence of fecal and flatus incontinence. We aimed to assess the clinical outcome of these repairs when done by the overlapping sphincteroplasty technique with reconstruction of the internal anal sphincter and perineum. Between August 2005 and December 2006, all grades 3 and 4 obstetric anal sphincter tears in our department were repaired by a reconstructive pelvic surgeon, primarily using the overlapping sphincteroplasty technique with reconstruction of the internal anal sphincter and perineum. All women were followed every 6 months using the Colorectal Anal Distress Inventory and Pelvic Organ Prolapse/Urinary Incontinence Sexual Function Questionnaire, a physical examination of the anal sphincter, anal manometry, and transperineal anal sonography. There were 3,478 deliveries of which 22 (0.63%) anal sphincter tears were repaired in women aged 22-41 years. Two women were diagnosed with Royal College of Obstetricians and Gynecologists grade 3a, eight with grade 3b, nine with grade 3c, and three with grade 4 anal sphincter tears. Postoperatively, 21 patients attended the outpatient clinic, with an average follow-up time of 9.2+/-1.4 months. Only two women (9.5%) complained of flatus incontinence and fecal urgency and had mildly decreased anal sphincter squeeze pressure and a small sonographic anal sphincter defect. None of the women complained of fecal incontinence. Two women (9.5%) reported on transient perineal pain and one (4.8%) on transient dyspareunia. All other women were asymptomatic and had normal anal manometry and sonographic evaluation. Repair of obstetric anal sphincter tears using the overlapping sphincteroplasty technique with reconstruction of the internal anal sphincter and perineum seems to carry favorable clinical outcome and reduced risk for anal incontinence, perineal pain, and sexual dysfunction. PMID- 18385918 TI - The effects of oral calcitonin on bone collagen maturation: implications for bone turnover and quality. AB - Anti-resorptive strategies may affect bone collagen maturation differently depending on the mode of action. Orally administrated calcitonin resulted in a dose dependent inhibition of bone resorption but did not change bone collagen maturation. This may reflect aspects of bone quality. INTRODUCTION: The aim of the present study was to evaluate the effect of oral calcitonin on bone collagen maturation measured as the ratio between the degradation products of newly synthesized C-telopeptides of type I collagen (alphaalphaCTX) and mature isomerized betabetaCTX in postmenopausal women. METHODS: Participants were from a phase II study. A total of 168 postmenopausal women were included and treated with placebo, 0.15, 0.4, 1, or 2.5 mg calcitonin daily. The non-isomerized alphaalphaCTX and isomerized betabetaCTX were measured in 24-hour urine samples obtained at baseline, and after 1 day, 1 month and 3 months of therapy. RESULTS: Calcitonin, significantly and dose-dependently inhibited bone resorption by up to 50% as measured by alphaalphaCTX and isomerized betabetaCTX. Bone collagen maturation measured as the ratio between alphaalphaCTX and betabetaCTX remained unchanged during treatment with calcitonin. CONCLUSIONS: Calcitonin dose dependently and significantly reduced both alphaalphaCTX to betabetaCTX levels in urine without affecting the alphaalphaCTX to betabetaCTX ratio. This is in direct contrast to other anti-resorptive therapies, in which strong treatment-dependent effect on the endogenous age profile of bone has been observed. These data highlight that even though the treatments may have comparable effects on BMD, endogenous bone composition, which may be associated to bone quality, is strongly affected by the type of intervention, in which calcitonin display highly divergent effects from that of other anti-resorptives. PMID- 18385919 TI - Strontium ranelate does not stimulate bone formation in ovariectomized rats. AB - INTRODUCTION: Strontium ranelate (SrR) is suggested to function as a dual-acting agent in the treatment of postmenopausal osteoporosis with anti-resorptive and anabolic skeletal benefits. We evaluated the effects of SrR on the skeleton in ovariectomized (OVX) rats and evaluated the influence of dietary calcium. METHODS: Three-month old virgin female rats underwent ovariectomy (OVX, n = 50) or SHAM surgery (SHAM, n = 10). Four weeks post-surgery, rats were treated daily by oral gavage with distilled water (10 ml/kg/day) or SrR (25 or 150 mg/kg/day) for 90 days. Separate groups of animals for each dose of SrR were fed a low (0.1%) or normal (1.19%) calcium (Ca) diet. Static and dynamic histomorphometry, DXA, mu-CT, mechanical testing, and serum and skeletal concentrations of strontium were assessed. RESULTS: SrR at doses of 25 and 150 mg/kg/day did not increase bone formation on trabecular or periosteal bone surfaces, and failed to inhibit bone resorption of trabecular bone regardless of Ca intake. There were no improvements in bone mass, volume or strength with either dose of SrR given normal Ca. CONCLUSION: These findings demonstrate that SrR at dosages of 25 and 150 mg/kg/day did not stimulate an anabolic bone response, and failed to improve the bone biomechanical properties of OVX rats. PMID- 18385920 TI - Morphological studies of pulmonary arteriovenous shunting in a lamb model of superior cavopulmonary anastomosis. PMID- 18385921 TI - The limits of confidence: at what price a baby's life? PMID- 18385922 TI - A third course of indomethacin. PMID- 18385923 TI - Radiological anatomy of the sigmoid colon. AB - BACKGROUND: The variable incidence of sigmoid volvulus, which depends on the presence of an elongated sigmoid colon, suggests the possibility of variations in the length of the sigmoid colon. This study was undertaken among the three major population groups to prove this hypothesis. PATIENTS AND METHODS: Radiological films of patients of the three population groups (African, Indian and White) undergoing barium enema were reviewed. The stature was measured by the distance from T12 to L4. The collective length of the rectum and sigmoid colon as well as the entire colon was measured on the barium enema film using an opisometer. Measurement was from the upper border of the symphysis pubis to the upper border of the left iliac crest. The level of the apex of the sigmoid colon loop and its redundancy were also assessed. RESULTS: There were 109 patients (61 females) undergoing barium enema (39 Africans, 49 Indians, and 21 Whites). For the entire group the T12-L4 distance was 16.6 +/- 2.2 cm and the entire colon length was 133 cm (range 88-262 cm) and was significantly longer among African patients (P = 0.003). The combined length of the rectum and sigmoid colon was 48.8 +/- 15.7 cm (Africans 60.9 +/- 14.4 cm, Indians 41.3 +/- 12.2 cm and Whites 44 +/- 11.6 cm). The sigmoid colon was significantly more redundant in Africans (90%), compared to Indians (25%) and Whites (24%) (P = 0.003 for Indians and P = 0.048 for whites). The apex of the sigmoid colon reached L1-L3 in 54% among Africans, 6% among Indians and in 10% among Whites (10%). CONCLUSION: African patients had the longest combined length of the rectum and sigmoid colon translating into a long sigmoid colon. They also had the highest number of redundant sigmoid colon. This may explain the high incidence of sigmoid volvulus in African patients. PMID- 18385925 TI - Implantation of episcleral electrodes via anterior orbitotomy for stimulation of the retina with induced photoreceptor degeneration: an in vivo feasibility study on a conceptual visual prosthesis. AB - BACKGROUND: A visual prosthesis is a conceptual device designed to harnesses the function of residual afferent neurons in the visual pathway to produce artificial vision. Such implant, when applied to stimulate the vitreous surface of the retina, has proven feasible in producing the perception of light in both animals and humans. However the practicality of such device has been challenged by the difficulty of surgical access and the risks of damaging the neuroretina. Positioning a visual implant over the scleral surface of the eye could present a safer alternative but this stimulation modality has not been tested in diseased retinas and little is known about the altered electrophysiological properties of the retina in influencing the feasibility of such approach. METHODS: Experimental photoreceptor degeneration was induced in four pigmented rabbit eyes with systematic administration of a retinotoxic agent, sodium iodate. A multielectrode array was implanted onto the surface of the sclera to target the central and peripheral parts of the retina via an anterior orbitotomy approach. The efficacy of retinal stimulation was assessed by recording electrical evoked potential over the primary visual cortex. FINDINGS: The electrical evoked potentials were obtained from both injected and control eyes. The charge density thresholds were found to be similar in both groups and were below the bioelectric safety limit. Spatially differentiated cortical activation profiles were obtained from the central and peripheral retina and the pattern of activation corresponded to the retinotopography of the rabbit primary visual cortex. CONCLUSION: This study proves that episcleral stimulation of the retina is a feasible alternative to intraocular approaches for the development of a visual prosthesis for retinas with photoreceptor loss. PMID- 18385924 TI - Topography and location of the depressor anguli oris muscle with a reference to the mental foramen. AB - The labiomandibular fold (LMF) is the area of the face that extends from the mouth corner to the mandibular border, and its prominence tends to increase with age. The LMF can be formed by the medial or lateral border of the depressor anguli oris (DAO). The aim of this study was to demonstrate the topographical anatomy between the DAO and mental foramen, thereby providing critical information for the safest and most effective site at which to inject botulinum toxin type A (BTX-A). Thirty-four hemifaces from Korean adult cadavers were dissected. The maximum width between the medial borders of the bilateral DAO, parallel to the intercheilion horizontal line, was 59.9 +/- 4.6 (mean +/- SD) mm below the lower lip. The minimum width between the medial borders of the attachment of bilateral DAO was 29.7 +/- 4.8 mm at the mandibular border. The mental foramen was located in the middle third from the cheilion to the mandibular border in 28 cases (90.3%), and it was mostly confined within the DAO muscle coverage in 21 cases (67.7%). The buccal branch of the facial nerve entered through the middle third of the lateral border of DAO and then distributed. Concomitantly, the marginal mandibular branch of the facial nerve entered through the lower third of the lateral border of DAO in 17 cases (60.7%). These results represent additional reference data for identifying the position of the mental foramen on the facial skin, and will be useful for providing criteria for the most effective site for injecting BTX-A when treating the LMF. PMID- 18385926 TI - Serum levels of S100B are decreased in chronic starvation and normalize with weight gain. AB - S100B protein is mainly synthesized in glial cells and modulates the balance between cell proliferation and differentiation in neurons and glial cells. However, S100B is not CNS-specific since its production was detected in numerous non-cerebral tissues e.g. adipocytes. In this study we investigated the influence of chronic fasting and subsequent weight gain on serum levels of S100B in patients with anorexia nervosa. We found that nutritional status was an important factor influencing serum levels of S100B. PMID- 18385927 TI - Rabbit hemorrhagic disease virus poly(A) tail is not essential for the infectivity of the virus and can be restored in vivo. AB - The precise role of the poly(A) tail at the 3'-end of the calicivirus RNA genome is unknown. To study the relationship between the presence of the poly(A) tail and the infectivity and replication of rabbit hemorrhagic disease virus (RHDV), mutants of an infectious cDNA clone of RHDV were constructed, and RK13 cells were transfected with transcripts from these mutants. Transcripts with and without a poly(A) had a fairly similar ability to infect and replicate, suggesting that a long 3'-terminal poly(A) is not essential for infectivity and replication. RT-PCR with specific primers, using viral RNA recovered from RK13 cells transfected with poly(A)-deficient RNA transcripts, showed that the poly(A) tail was restored in vivo. PMID- 18385929 TI - Lehmann effect in a compensated cholesteric liquid crystal: experimental evidence with fixed and gliding boundary conditions. AB - In a recent letter (Europhys. Lett. 80, 26001 (2007)), we have shown that a compensated cholesteric liquid crystal (in which the macroscopic helix completely unwinds) may be subjected to a thermomechanical torque (the so-called Lehmann effect), in agreement with previous findings of Eber and Janossy (Mol. Cryst. Liq. Cryst. Lett. 72, 233 (1982)). These results prove that one must take into account the chirality of the molecules and the absence of inversion symmetry at the macroscopic scale when deriving the constitutive equations of the phase at the compensation temperature. In this paper, we present the details of our experimental work and a new experiment performed in a sample treated for planar gliding anchoring. The latter experiment, coupled with a numerical simulation, supports the existence of a thermomechanical coupling in a compensated cholesteric. PMID- 18385928 TI - Novel halophilic aerobic anoxygenic phototrophs from a Canadian hypersaline spring system. AB - The first enumeration of cultivable obligately aerobic phototrophic bacteria from a terrestrial saline spring was accomplished in the East German Creek system (salinity approximately 6%), near Lake Winnipegosis, Manitoba, Canada. Occurring at densities up to 3.3x10(7) CFU/ml of sample, aerobic phototrophs comprised 15 36% of the total cultivable bacterial population in the diatom- and chlorophyte dominated aerobic microbial mats. Many of the representative strains isolated for phenotypic characterization and phylogenetic analysis possessed <96% 16S rDNA sequence overlap with published species, including an obligately aerobic phototrophic gammaproteobacterium displaying only 92.9% 16S rDNA sequence similarity to Congregibacter litoralis. The springs yielded the most highly halotolerant aerobic anoxygenic phototroph yet recorded, strain EG11, which grew with 26% NaCl. PMID- 18385930 TI - The absence of physical-aging effects on the surface relaxations of rubbed polystyrene. AB - Extensive experimental results are presented to reveal the relaxations of polystyrene surface deformed by rubbing with a velvet cloth. We found that surface topographic features, such as ditches and ridges created by rubbing, relax at temperatures at about 20( degrees ) C below the bulk glass transition temperature of the polystyrene for a molecular weight of 442 kg/mol, even though we estimate the Laplace Pressure driving the relaxation to be 1/500 of the yield limit. The relaxation is independent of the thermal history before the rubbing process, and post rubbing thermal history below 55( degrees ) C . In other words, physical-aging processes at 23( degrees ) C for up to 7 days and at 50( degrees ) C for 2 days, which would have drastic effects on the relaxations of bulk polymers, have little effects on the relaxations of rubbed surfaces. This is consistent with the mobility enhancement in the surface layer previously reported in the literature. PMID- 18385931 TI - Structural transitions of encapsidated polyelectrolytes. AB - Conformations and structural transitions of polyelectrolytes strictly confined onto a spherical 2D surface have been investigated by scaling descriptions based on physical arguments concerning polyelectrolyte adsorption onto planar surface and liquid crystals as well as by Monte Carlo simulations using a bead-spring model with short-range and electrostatic repulsions. In case of the electrostatic screened regime, a disordered-ordered (spiral) transition at increasing persistence length of the chain was found. It was predicted that the transition occurred when the persistence length is comparable with the mean spacing between adjacent strands of the ordered chain. The presence of a non-screened electrostatic repulsion led to a more complex behavior with i) a re-entrant order disorder transition and ii) a tennis ball texture as an additional smectic/nematic structure. The various competing structures given by the theory were recovered by the Monte Carlo simulations, which also indicated that the tennis ball texture was favored over the spiral structure by the long-range interactions for semi-flexible chains. PMID- 18385932 TI - Adhesion and fingering in the lifting Hele-Shaw cell: role of the substrate. AB - The lifting Hele-Shaw cell (LHSC) is used to study adhesion as well as viscous fingering. In the present paper we report a series of observations of development of the interface for different viscous fluids, both Newtonian and non-Newtonian, in a LHSC operated at a constant lifting force. Glass and perspex are used as the plates in two different sets of experiments. The objectives are 1) to measure the time required to separate the plates as a function of the lifting force and 2) to note the force above which viscous fingering appears. We find that for the Newtonian fluids, the plate separation time follows a universal power law with the lifting force, irrespective of fluid and substrate. The non-Newtonian fluids too, with proper scaling obey the same power law. The appearance of fingering, however, depends on the properties of the fluid as well as the substrate. We suggest a modified form of the capillary number which controls the onset of fingering; this new quantity, termed the "fingering parameter" involves the dielectric constants of the substrate and fluid in addition to the viscosity and surface tension. PMID- 18385933 TI - Topiramate in the prevention of pediatric migraine: literature review. AB - Pediatric migraine is a disabling condition, which can cause a significant impact on quality of life. Currently, no drugs have been approved by the FDA for its preventive treatment. Our aim was to review the medical literature concerning the efficacy and tolerability of topiramate in the prophylactic treatment of migraine in children and adolescents. A total of five papers were reviewed: two randomized controlled trials (RCTs), a post-hoc subset analysis of adolescents who had been included in three RCTs carried out on adults and two open studies. Topiramate has been proven to reduce headache frequency and the accompanying disability. The frequency of side effects varied considerably among studies, the most frequent being weight loss, anorexia, abdominal pain, difficulties in concentrating, sedation and paresthesia. Since these adverse events, although often transitory, may be distressing for the child, we strongly recommend to assess the disability caused by the migraine episodes before deciding to initiate a prophylactic treatment. Nevertheless, dropout rates due to side effects in the studies were very low. PMID- 18385934 TI - Characterisation of TNF-related apoptosis-inducing ligand in peripheral blood in patients with primary biliary cirrhosis. AB - Apoptosis plays a pivotal role in portal tract damage of primary biliary cirrhosis (PBC). Tumour necrosis factor-related apoptosis inducing ligand (TRAIL) is an apoptotic inducer, and it has been reported that the expression of TRAIL receptors is up-regulated by increased bile acid level and the serum level of soluble TRAIL (sTRAIL) is elevated in PBC patients. In the present study, we investigated the association of TRAIL in peripheral blood with the pathogenesis of PBC and chronic hepatitis B. The expression levels of TRAIL mRNA and protein on leukocytes and sTRAIL in plasma from 27 patients with PBC, 25 with CHB and 30 healthy controls were determined respectively by real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR), flow cytometry (FCM) and ELISA. The expression levels of TRAIL mRNA and protein on leukocytes and plasma sTRAIL were all up-regulated in the patients with PBC and CHB compared to controls. In the two diseased groups, TRAIL mRNA showed significant correlation of both membrane-bound TRAIL (mTRAIL) on monocytes and plasma sTRAIL. So did plasma TNF-alpha. In PBC patients, mTRAIL and sTRAIL correlated well with gamma-glutamyltransferase and alkaline phosphatase, but not with aspartate aminotransferase and alanine amino-transferase. The opposite case was found in CHB patients. These results suggested that both mTRAIL and sTRAIL might be involved in the development and progression of PBC and CHB in humans, but the mechanisms might be different. PMID- 18385935 TI - The value of antinuclear antibodies in primary biliary cirrhosis. AB - OBJECTIVE: Although autoantibodies have been used for the diagnosis of primary biliary cirrhosis (PBC), their role has not been clarified. In this study, we try to explore the value of gp210 antibody and anti-centromere antibodies (ACA) in PBC. METHODS: Anti-gp210 and ACA were tested in 140 PBC patients by ELISA and indirect immunofluorescence respectively. Their association with clinical, pathological data and prognosis was analysed. RESULTS: 30.5% of PBC patients had positive anti-gp210 antibody and 29.2% had ACA. The anti-gp210 antibody positive group had higher Mayo risk scores and lower serum albumin levels compared to the negative one. Patients with positive anti-gp210 antibody were more likely to develop hepatic failure (p<0.05, OR=9.8460, 95% CI: 1.067-90.901) than patients with negative anti-gp210 antibody. More patients with positive ACA developed portal hypertension than patients with negative ACA (p<0.05, OR=9.259; 95% CI: 1.027-88.410). Furthermore, concurrent Sjogren's syndrome (SjS) and PBC was significantly more likely in the ACA positive group than in the negative ones (68.4% in ACA positive group, 20.7% in ACA negative group p<0.001). CONCLUSIONS: Both anti-gp210 antibody and ACA are related to severe disease course and poor prognosis. For PBC patients with positive ACA, further examinations should be made to detect underlying SjS. PMID- 18385936 TI - Exon 33 T/T genotype of the thyroglobulin gene is a susceptibility gene for Graves' disease in Taiwanese and exon 12 C/C genotype protects against it. AB - This study investigates whether Tg gene polymorphisms can be associated with Graves' disease (GD) in a Taiwanese population and identifies potential polygenic susceptive genes for GD. The findings of such a study may have important implications for prognostic prediction and treatment of GD. We performed case control association studies for the 3 discovered Tg single nucleotide polymorphisms (SNPs) (E10, E12, E33) in 215 GD patients and 141 controls. The three SNPs were identified within the Tg gene. These SNPs were analysed by a fluorescent-based restriction fragment length polymorphism method (RFLP) and PCR. The genotype and allele frequencies at E10SNP158, E12SNP and E33SNP in GD patients were compared with those of the controls. In addition, we analysed the interactions between these SNPs and the clinical and laboratory variables. We found a significant difference in the T/T genotype of E33SNP and G/G genotype of E12SNP compared with the control group (p<0.001). We also found the E33SNP T/T genotype to be positively associated with development of GD, whereas the E12SNP G/G genotype protected it. PMID- 18385937 TI - Genetic association of interleukin-1 haplotypes with gastritis and precancerous lesions in North Indians. AB - BACKGROUND: We evaluated the association of functional variants of IL-1 genes with the development of gastritis and precancerous lesions, which are known to be influenced by inflammatory response against Helicobacter pylori. METHODS: After upper gastrointestinal (GI) endoscopy, 120 patients with gastritis were tested for H. pylori infection using rapid urease test, modified Giemsa staining and IgG anti-CagA ELISA. All patients and 243 healthy controls were genotyped for IL-1B ( 511 C/T) and IL-IRN (VNTR) genes using PCR-RFLP/PCR. RESULTS: IL-1B: (-511 C/T) genotype/allele were not associated with gastritis. IL-1RN 1/2 genotype carriers had susceptibility to gastritis (p=0.025, OR=1.7). Individuals with the IL-1RN 1/1 genotype (p=0.05, OR=0.65) and IL-1B -511*T-IL-1RN *1 haplotype were at low risk for gastritis (p=0.043, OR=0.72). High secretor haplotype combinations (C1 /T2+, C1-T1+ and T1+/T2+) did not influence neutrophilic infiltration, glandular atrophy or intestinal metaplasia. CONCLUSIONS: We identified that individuals with the IL-1RN 1/2 genotype had increased risk for gastritis. IL-1B -511*T-IL 1RN *1 (T1) haplotype carriers were at decreased risk for gastritis and no significant association was observed for precancerous lesions in North Indians. PMID- 18385939 TI - Expression of soluble, biologically active recombinant human tumstatin in Escherichia coli. AB - Tumstatin, a 28-kDa C-terminal fragment of collagen IV, is a potent anti angiogenic protein and inhibitor of tumour growth. Recombinant tumstatin was prepared from Escherichia coli deposited as insoluble, inactive inclusion bodies. In the present study, we produced soluble and biologically active recombinant human tumstatin in E. coli by the coding region of tumstatin being linked to the 3'-end of the maltose-binding protein (MBP) gene. The fusion protein was expressed as the soluble form after induction by isopropylthio-beta-D-galactoside (IPTG). MBP-tumstatin was purified by amylose affinity chromatography. MBP can be removed by digestion with factor Xa. Expression could represent 20% of the total soluble protein in E. coli, allowing approximately 8.6 mg of highly purified protein to be obtained per litre of bacterial culture. The purified tumstatin specifically inhibited the proliferation of endothelial cells in a dose-dependent manner. Annexin V-FITC apoptotic assay showed that recombinant tumstatin induced significant increase of apoptotic endothelial cells after 20 h of exposure to 20 microg/ml tumstatin, and when tumstatin was incubated on the chicken embryo, chorioallantoic membrane at doses of 1-15 microg, there was a dramatic decrease in the microvasculature allantoids of chicken embryos neovascular vessel test in vivo demonstrated that tumstatin treatment at doses of 1-15 microg gives rise to dramatically decrease the number of neovascular vessel. Our study provides a feasible and convenient approach to produce soluble and biologically active tumstatin. PMID- 18385938 TI - Sensitivity to cisplatin-induced mutations and elevated chromosomal aberrations in lymphocytes from sickle cell disease patients. AB - Sickle cell disease (SCD) is an inherited disorder caused by a single nucleotide substitution in the beta-globin gene. The clinical heterogeneity observed in SCD patients has been attributed to environmental and genetic factors. The patients are subjected to increased oxidative stress, particularly during vaso-occlusive crises and acute chest pain. Another possible cause of oxidative stress in SCD is the high concentration of iron in the patients' plasma. The increase in oxidative stress could be a relevant risk factor for mutagenesis and carcinogenesis. Studies on the frequency of basal chromosomal aberrations in cultured lymphocytes from SCD patients have not been reported so far. In order to contribute to the understanding of the role of the different biomarkers and their relationship with the extremely variable clinical manifestation of SCD, we investigated the frequency of chromosome damage in peripheral lymphocytes from sickle cells patients and healthy controls. We found an increased frequency of chromosome damage and percentage of aberrant metaphases in these patients when compared with control subjects, even at basal values (p<0.05). In the cytogenetic sensitivity assay, the results showed that these patients presented a marked decrease in the mitotic index values compared with healthy controls. Cisplatin-induced chromosomal damage in lymphocytes from these patients was significantly higher than the frequency measured in healthy controls. The results obtained in the present study showed that more investigations are needed in order to elucidate the susceptibility to genomic instability of SCD patients. PMID- 18385940 TI - Effect of the antioxidant alpha-lipoic acid on apoptosis in human umbilical vein endothelial cells induced by high glucose. AB - High glucose plays an important role in the pathogenesis of atherosclerosis. In this study, we assessed the effects of high glucose on human umbilical vein endothelial cell (HUVEC) apoptosis. Additionally, we investigated whether alpha lipoic acid, an antioxidant, prevents high glucose-induced apoptosis of HUVECs. HUVECs were treated with high glucose in the presence or absence of alpha-lipoic acid. Treatment of HUVECs with high glucose changed cell morphology and induced DNA fragmentation, leading to apoptosis. Apoptosis was induced by high glucose in a dose-and time-dependent fashion. High glucose markedly elevated Bax, and decreased NF-kappaB and Bcl-2 expression. Most importantly, pretreatment with alpha-lipoic acid protected against high glucose-induced apoptosis in the endothelial cells. alpha-Lipoic acid significantly promoted the expression of NF kappaB while decreasing the expression of Bax and the activities of caspase-3 and 9 without significantly affecting the Bcl-2 level. Our data suggest that high glucose induces apoptosis in endothelial cells. alpha-Lipoic acid effectively attenuates high glucose-induced endothelial cell apoptosis. These findings provide new perspectives on the role of alpha-lipoic acid in cardiovascular disease. PMID- 18385941 TI - Relation of plasma protein oxidation parameters and paraoxonase activity in the ageing population. AB - The incidence of atherosclerosis increases with age. Oxidative changes in proteins and lipids are considered to be among the molecular mechanisms leading to endothelial dysfunction. Paraoxonase (PON1) is exclusively associated with high-density lipoprotein (HDL) and protects both HDL and low-density lipoprotein (LDL) from oxidation. PON1 has two cysteine residues for its antioxidant function. We investigated the relation between PON1 activity and protein oxidation parameters such as protein hydroperoxides (P-OOH), protein carbonyl (PCO), total thiol (T-SH) and advanced oxidation protein products (AOPP). Our study also covered other oxidative stress parameters such as oxidised LDL (oxLDL) and superoxide dismutase activity in the plasma of young, middle-aged and elderly individuals. PON1 activity of elderly and middle-aged individuals was decreased significantly compared with that in the young group. oxLDL levels of elderly individuals were increased significantly compared with those of both the young and middle-aged individuals. P-OOH, PCO and AOPP levels of the elderly and middle aged individuals were higher compared with those of the young. On the other hand, T-SH levels of the elderly and middle-aged individuals were lower compared with those of the young. Side by side with the decrease in the T-SH levels in the middle-aged and elderly groups as compared to the young, the increase we have observed in other protein oxidation parameters in the groups leading to decreasing PON1 activity might, we think, create a predisposition to atherosclerosis. PMID- 18385942 TI - Clinical significance of autoantibodies to inner ear antigens in sera of patients with systemic diseases. PMID- 18385943 TI - The JNK/AP1/ATF2 pathway is involved in H2O2-induced acetylcholinesterase expression during apoptosis. AB - We show that H2O2 increases acetylcholinesterase (AChE) expression via transcriptional activation through c-Jun N-terminal kinase (JNK), since the JNK inhibitor SP600125, but not the extracellular signal-regulated kinase (ERK) pathway inhibitor PD98059 or p38 kinase inhibitor SB203580, attenuated H2O2 induced AChE expression and its promoter activity. Overexpression of hemagglutinin (HA)-JNK increases H2O2-induced AChE expression and its promoter activity, whereas the dominant negative mutant form of JNK suppressed H2O2 induced AChE expression and promoter activity. Mutation analysis indicates that the major response elements for JNK in the AChE promoter are the AP1-like element (TGAGTCT) site, located within the -1565/-1569 region of the AChE promoter, and the ATF2 element (CCACGTCA), within the -2185/-2177 region. The AP1-like element binds to the transcription factors, c-jun and ATF2, while the ATF2 element binds mainly ATF2. Taken together, our results strongly suggest that H2O2 induces AChE expression via the JNK/AP1/ ATF2 signaling pathway. PMID- 18385944 TI - Regulation of phagocyte migration and recruitment by Src-family kinases. AB - Src-family kinases (SFKs) regulate different granulocyte and monocyte/macrophage responses. Accumulating evidence suggests that members of this family are implicated in signal transduction pathways regulating phagocytic cell migration and recruitment into inflammatory sites. Macrophages with a genetic deficiency of SFKs display marked alterations in cytoskeleton dynamics, polarization and migration. This same phenotype is found in cells with either a lack of SFK substrates and/or interacting proteins such as Pyk2/FAK, c-Cbl and p190RhoGAP. Notably, SFKs and their downstream targets also regulate monocyte recruitment into inflammatory sites. Depending on the type of assay used, neutrophil migration in vitro may be either dependent on or independent of SFKs. Also neutrophil recruitment in in vivo models of inflammation may be regulated differently by SFKs depending on the tissue involved. In this review we will discuss possible mechanisms by which SFKs may regulate phagocytic cell migratory abilities. PMID- 18385945 TI - Cystoid macular oedema within 24 h after a single application of latanoprost. AB - PURPOSE: To describe a patient who developed cystoid macular oedema (CMO) within 24 h after a single application of Latanoprost. METHODS: Observational case report. RESULTS: A 77-year-old man who had a previous cataract operation with vitreous loss in his left eye developed CMO 8 h after instillation of one drop of latanoprost. Five days after discontinuation of latanoprost the CMO resolved almost completely. Both the CMO and its resolution were documented with fluorescein angiography (FA) and optical coherence tomography (OCT). CONCLUSIONS: Latanoprost can lead to CMO even after a single application, as demonstrated in our case by FA and, in particular, by OCT. Caution is advised when administering the drug in high-risk eyes. PMID- 18385946 TI - Changing trends in the clinical course and outcome of bacterial keratitis at King Khaled Eye Specialist Hospital. AB - OBJECTIVE: To evaluate changes in the predisposing factors and causative organisms, clinical course, and outcome of bacterial keratitis at King Khaled Eye Specialist Hospital (KKESH) between 1995 and 2005. METHODS: Retrospective review of 78 and 103 cases of culture-positive bacterial keratitis admitted to KKESH in 1995 and 2005. Main outcome measures Microbiological cure, visual outcome. RESULTS: There was little change in the predisposing factors for bacterial keratitis, initial clinical severity, or the microbiological profile between 1995 and 2005. While similar topical fortified antibiotic therapy regimens were used in both 1995 and 2005, there was a significant increase in the use of topical corticosteroids in 2005 (P < 0.001), either alone or in combination with medroxyprogesterone 1% and doxycycline. There were significant improvements in the percentage of eyes achieving microbiological cure with medical therapy alone (76.0 vs. 92.2%, P = 0.002) or in combination with surgical intervention (92.4 vs. 100.0%; P = 0.005). The percentage of eyes requiring enucleation or evisceration due to treatment failure declined significantly from 7.6 to 0% (P = 0.006). There was a significant improvement in the percentage of eyes achieving a final visual acuity > or =20/40 (10.1 vs. 29.1%, P = 0.001). CONCLUSIONS: Significantly better clinical outcomes were achieved in 2005 compared to 1995. Increased emphasis on management of inflammation in conjunction with the infectious process may have contributed to the improved outcomes. PMID- 18385947 TI - Investigation of structures and properties of cyclic peptide nanotubes by experiment and molecular dynamics. AB - In order to investigate the structures and properties of cyclic peptide nanotubes of cyclo[(-D: -Phe-L: -Ala)( n = 3,4,5,6)-], cyclo[(-D: -Phe-L: -Ala)( n = 4)-] was synthesized and self-assembled to nanotubes, and its structure and morphology of the nanotube were characterized by mass spectrometry (MS), fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM). On the basis of these experimental results, the structures of cyclo[(-D: -Phe-L: -Ala)( n = 3,4,5,6)-] were characterized by molecular dynamics. In addition, the motion behaviors of H(2)O molecules in nanotubes were investigated by molecular dynamics using a COMPASS force field. Experimental results show that cyclo[(-D: -Phe-L: Ala)( n = 4)-] peptides self-assemble into nanotube bundles. Molecular modeling results indicate that cyclic peptide nanotubes with n = 3, 4, 5 and 6 are very stable; these nanotubes have internal diameters of 5.9 A, 8.1 A, 10.8 A and 13.1 A and outer diameters of 18.2 A, 21.7 A, 23.4 A and 25.9 A respectively. Modeling results demonstrate that H(2)O molecules move in cooperation in single nanotube and they diffuse in one dimension, but they did not diffuse unilaterally due to the antiparallel ring stacking arrangement. PMID- 18385948 TI - The effects of characteristics of substituents on toxicity of the nitroaromatics: HiT QSAR study. AB - The present study applies the Hierarchical Technology for Quantitative Structure Activity Relationships (HiT QSAR) for (i) evaluation of the influence of the characteristics of 28 nitroaromatic compounds (some of which belong to a widely known class of explosives) as to their toxicity; (ii) prediction of toxicity for new nitroaromatic derivatives; (iii) analysis of the effects of substituents in nitroaromatic compounds on their toxicity in vivo. The 50% lethal dose concentration for rats (LD50) was used to develop the QSAR models based on simplex representation of molecular structure. The preliminary 1D QSAR results show that even the information on the composition of molecules reveals the main tendencies of changes in toxicity. The statistic characteristics for partial least squares 2D QSAR models are quite satisfactory (R2 = 0.96-0.98; Q2 = 0.91 0.93; R2 (test) = 0.89-0.92), which allows us to carry out the prediction of activity for 41 novel compounds designed by the application of new combinations of substituents represented in the training set. The comprehensive analysis of toxicity changes as a function of substituent position and nature was carried out. Molecular fragments that promote and interfere with toxicity were defined on the basis of the obtained models. It was shown that the mutual influence of substituents in the benzene ring plays a crucial role regarding toxicity. The influence of different substituents on toxicity can be mediated via different C-H fragments of the aromatic ring. PMID- 18385949 TI - Simulated microgravity activates apoptosis and NF-kappaB in mice testis. AB - Microgravity is known to have significant effect on all aspects of reproductive function in animal models. Recent studies have also shown that microgravity induces changes at the cellular level, including apoptosis. Our effort here was to study the effect of simulated microgravity on caspase-8 and the caspase-3 activities, the effectors of the apoptotic pathway and on the transcription factor NF-kappaB a signaling molecule in mouse testis. Morey-Holton hind limb suspension model was used to simulate microgravity. Caspase-8 and 3 fluorometric assays were carried out and HLS mice testis exhibited a 51% increase in caspase-8 and caspase-3 compared to the controls. A sandwich ELISA-based immunoassay was carried out for detection of NF-kappaB which again significantly increased in the test mice. Testosterone levels were measured using an ELISA kit and in HLS mice the decrease was significant. There was also a significant decrease in testis weight in the test mice. Simulated microgravity activates caspase 8, 3 and NF kappaB necessary to stimulate the apoptotic pathway in mice testis. This may account for the drop in testis weight and testosterone level further affecting testicular physiology and function. PMID- 18385950 TI - The liquid-ordered phase in sphingomyelincholesterol membranes as detected by the discrimination by oxygen transport (DOT) method. AB - Membranes made from binary mixtures of egg sphingomyelin (ESM) and cholesterol were investigated using conventional and saturation-recovery EPR observations of the 5-doxylstearic acid spin label (5-SASL). The effects of cholesterol on membrane order and the oxygen transport parameter (bimolecular collision rate of molecular oxygen with the nitroxide spin label) were monitored at the depth of the fifth carbon in fluid- and gel-phase ESM membranes. The saturation-recovery EPR discrimination by oxygen transport (DOT) method allowed the discrimination of the liquid-ordered (l(o)), liquid-disordered (l(d)), and solid-ordered (s(o)) phases because the bimolecular collision rates of the molecular oxygen with the nitroxide spin label differ in these phases. Additionally, oxygen collision rates (the oxygen transport parameter) were obtained in coexisting phases without the need for their separation, which provides information about the internal dynamics of each phase. The addition of cholesterol causes a dramatic decrease in the oxygen transport parameter around the nitroxide moiety of 5-SASL in the l(o) phase, which at 50 mol% cholesterol becomes approximately 5 times smaller than in the pure ESM membrane in the l(d) phase, and approximately 2 times smaller than in the pure ESM membrane in the s(o) phase. The overall change in the oxygen transport parameter is as large as approximately 20-fold. Conventional EPR spectra show that 5-SASL is maximally immobilized at the phase boundary between regions with coexisting l(d) and l(o) phases or s(o) and l(o) phases and the region with a single l(o) phase. The obtained results allowed for the construction of a phase diagram for the ESM-cholesterol membrane. PMID- 18385951 TI - Kinetic models for stochastically modified ionic channels. AB - Ionic channels form pores in biomembranes. These pores are large macromolecular structures. Due to thermal fluctuations of countless degrees-of-freedom of the biomembrane material, the actual form of the pores is permanently subject to modification. Furthermore, the arrival of an ion at the binding site can change this form by repolarizing the surrounding aminoacids. In any case the variations of the pore structure are stochastic. In this paper, we discuss the effect of such modifications on the channel conductivity. Applying a simple kinetic description, we show that stochastic variations in channel properties can significantly alter the ionic current, even leading to its substantial increase or decrease for the specific matching of some time-scales of the system. PMID- 18385952 TI - Lysosomes of the liver and the duodenal enterocytes, a site of handling of two rare earths. Ultrastructural and microanalytical study. AB - The frequent use of some trace elements such as gadolinium and terbium in medicine and modern industries make us worry about their behavior in the organism. In this work, we study the intracellular localization in the liver and in the intestinal mucosa of two rare earths, gadolinium and terbium, after intraperitoneal and intragastric administration. Three methods of observation and microanalysis were used: conventional transmission electron microscopy, secondary ion mass spectrometry, and electron probe microanalysis. After intraperitoneal administration, gadolinium and terbium were detected with phosphorus in lysosomes of hepatocytes and Kuppfer cells and in territories near to biliary canalicule. One hour after intragastric administration, gadolinium and terbium were concentrated in lysosomes of the apical part of duodenal enterocytes. No gadolinium or terbium was detected in duodenum 4 days after administration. After intragastric administration, the microanalytical techniques failed to detect gadolinium or terbium in liver whatever the time of sampling. This mechanism of concentration-precipitation in the lysosomes of enterocytes limits the diffusion through the digestive barrier of foreign elements and then permits their elimination with apoptotic cells in the intestinal lumen. Some of these elements may be toxic, and none of them have a recognized physiological function. The intestinal mucosa plays an important role in the protection of the organism against the invasion of foreign elements. PMID- 18385953 TI - The long-term effects of lamivudine treatment in patients with HBeAg-negative liver cirrhosis. AB - INTRODUCTION: In hepatitis B virus (HBV)-related liver cirrhosis, patients with HBV replication show a higher mortality rate than those without. We aimed to investigate the long-term effects of lamivudine on HBV DNA suppression, Child Pugh score, and survival in patients with hepatitis Be antigen (HBeAg)-negative liver cirrhosis. METHODS: Sixty-eight patients (51 male, 17 female) diagnosed with HBV-positive liver cirrhosis, who were monitored by the hepatology and liver transplantation outpatient clinics of our hospital between June 1999 and May 2007, were included in the study. Lamivudine (100 mg/day) was administered orally. Follow-up visits were scheduled monthly during the first 3 months, and every 3 months thereafter. Complete blood count, haemostasis, biochemistry (aspartate aminotransferase [AST], alanine aminotransferase [ALT], amylase, urea, creatinine, total bilirubin, direct bilirubin, total protein, albumin), and alpha foetoprotein were recorded every 3 months. HBV DNA levels, abdominal ultrasound and the Child-Pugh score were evaluated every 6 months. RESULTS: Sixty-eight patients (mean age, 52.05+/-12.6 years) were monitored for 49.51+/-18.51 months. Basal ALT, HBV DNA levels and Child-Pugh scores were 103.9+/-73.9 IU/ml, 4133+/ 121,94 IU/ml, and 7.6+/-2.4, respectively. The ALT normalisation was 59.7% during the first year, 68.2% during the second year and 44.4% during the fifth year. There was a significant decrease in Child-Pugh scores in the first 3 follow-up years when compared with the baseline score (P<0.05). During the treatment, HBV DNA positivity and YMDD mutations were determined in 20 of 68 (29.4%) patients at 46+/-17.9 months. Nine patients (13.2%) developed hepatocellular carcinoma at 44.8+/-21.5 months. Thirteen patients (19.1%) died during the treatment due to liver failure or variceal bleeding. CONCLUSION: Lamivudine is beneficial in patients with HBeAg-negative liver cirrhosis in terms of improvement in liver function and enhancement of survival and quality of life. An HBV DNA suppressive effect and improvement in Child-Pugh score were seen especially in the first years. It is important to be aware of YMDD mutation early, as addition of new antivirals is necessary to overcome unwanted results of the mutation. PMID- 18385954 TI - A new generation of self-etching adhesives: comparison with traditional acid etch technique. AB - OBJECTIVE: The aim of this study was to determine the shear bond strength (SBS), etching pattern and depth, and debonding performance of several market-leading, self-etching (SE) adhesives primarily used in restorative dentistry (iBond, Clearfil S(3) Bond, Clearfil Protect Bond, AdheSE, XenoIII), two experimental self-etching adhesives (exp. Bond 1, exp. Bond 2) and one experimental self etching cement (SE Zement) used with and without prior phosphoric acid-etching, and to compare them to an orthodontic self-etching product (Transbond Plus SE Primer) and to traditional acid-etch technique (Transbond XT Primer, phosphoric acid) MATERIALS AND METHODS: All adhesives were applied on pumiced and embedded bovine incisors following the manufacturers' instructions. Then one bracket each (coated with Transbond XT composite) was bonded (n = 20). Transbond XT was polymerized for 20 s from the incisal and gingival sides using a halogen device positioned at a constant 5 mm from and a 45 degrees angle to the specimen. The specimens were stored in distilled water for 24 h at 37 degrees C before measuring SBS. The ARI (adhesive remnant index) for all specimens was determined from the sheared-off brackets of each. After conditioning, the surface texture was morphologically evaluated from scanning electron microscope (SEM) images, while the etching depth was determined using a confocal laser-scanning microscope (CLSM). All groups were tested for normal distribution and analyzed by applying ANOVA, Kruskal-Wallis or the t test. In addition, a Bonferroni correction was used. RESULTS: The median values of the SBS tests were: SE Zement 3.0 MPa, SE Zement preceded by phosphoric acid etching 11.2 MPa, experimental bond 1: 7.4 MPa, experimental bond 2: 5.6 MPa, iBond 8.1 MPa, Clearfil S(3) Bond 14.1 MPa, Clearfil Protect Bond 16.6 MPa, Clearfil SE Bond 15.9 MPa, AdheSE 16.0 MPa, XenoIII 16.1 MPa, Transbond SE Primer 20.7 MPa, acid-etching+Transbond XT Primer 21.0 MPa. With the exception of iBond, we observed no significant differences among the self-etching adhesives used in Restorative Dentistry or in comparison to the Transbond Plus SE Primer. No significant differences were apparent even when compared to the Transbond XT Primer after phosphoric acid-etching. Both experimental bonding agents and SE Zement without acid etching performed significantly worse than the products mentioned above, failing to demonstrate sufficient adhesive strength. SEM examination revealed less distinctive enamel etching patterns for self-etching products than for phosphoric acid-etching. CLSM analysis revealed etching depths between 0.5 and 20 microm depending on the product. When self-etching products were used, less residual composite remained on the enamel surface than after phosphoric acid-etching. CONCLUSIONS: All the adhesives tested are suitable for bonding orthodontic brackets and to reduce the risk of enamel fracture while minimizing etching depth, which in turns means less conditioning-related enamel loss. More development is needed to improve the etching performance of both experimental bonding agents and SE Zement. PMID- 18385955 TI - Sagittal space relations in the maxilla during molar eruption. AB - OBJECTIVE: The aim of this study was to measure the space relations in the maxilla during upper molar eruption, and to interpret associations. MATERIAL AND METHODS: 154 skulls ranging from infant to adult were measured on the basis of defined distances. The age of the skulls was determined by tooth development. The data were compared and qualitatively interpreted. RESULTS: We observed that, immediately after eruption of the primary dentition, there was nearly enough space in the upper jaw for the first permanent molar. Yet eruption does not occur until the age of 5 to 6 years, at the beginning of the first transition phase. As opposed to the first permanent molar, the second erupted as soon as there is enough space in the upper jaw. This tends to happen at the end of the second transition phase at ca. 12-13 years of age. We noted that there was almost enough space in the maxilla until the age of 17 to accommodate all the teeth in the upper jaw. According to our measurements, growth at the posterior edge of the maxilla also took place after eruption of the third molar. CONCLUSIONS: This leads us to consider that we can postpone the extraction of the upper third molars until after the 18th year, and to consider the possibility that the upper third molar may erupt correctly while the maxilla is still growing. It is precisely this trend that is significant in clinical practice, since maxillary wisdom teeth are often extracted prematurely due to an apparent lack of space. PMID- 18385956 TI - Cephalometric changes in the treatment of class III using the Frankel appliance. AB - AIM: In this retrospective clinical study, we aimed to evaluate skeletal changes following treatment of Class III malocclusion by means of the Function Regulator (FR-3) appliance. MATERIAL AND METHODS: The group of patients we treated with the FR-3 exclusively consisted of 56 subjects (20 boys and 36 girls). Fifteen subjects (7 boys and 8 girls) of approximately the same age and presenting almost identical characteristics of a Class III malocclusion of skeletal etiology served as a control group (between the ages of 7 and 15 years). Lateral cephalograms were compared using the occipital reference structures as proposed by Frankel. RESULTS: Cephalometric analysis showed that maxillary landmarks (point A, nasospinale), and the upper incisor (root included) did move significantly farther forward in the treated group than in the control group (p < 0.01). We also found that the gonial angle decreased by 7.17 degrees in the FR-3 group, as opposed to 2.07 degrees in the untreated group (p < 0.001). No significant differences were noted in mandibular-length growth (Ar-Pog, Ar-Go, Go-Pog). CONCLUSION: According to these study results, FR-3 treatment has an obvious effect on maxillary development, and affects the shape and, to a certain degree, the position of the mandible. PMID- 18385957 TI - Anchorage efficacy of palatally-inserted miniscrews in molar distalization with a periodontally/miniscrew-anchored distal jet. AB - AIM: As a clinical pilot study using the skeletonized, periodontally/miniscrew anchored Distal Jet appliance, this study aimed to verify the positional stability of the palatally-inserted paramedian miniscrews when subjected to loading for several months, hence to assess the efficacy of the supporting anchorage design. MATERIAL AND METHOD: Sixteen miniscrews (8-9 mm in length, 1.6 mm in diameter, polished surface) were inserted in the anterior region of the palate at paramedian locations. Once they had been in place for 1 week, skeletonized Distal Jets for bilateral molar distalization were anchored to the first premolars and necks of the miniscrews using composite. The appliances' coil spring systems were activated to a distalization force of 200-240 cN. The miniscrews were processed histologically after minimally-invasive explantation. RESULTS: Forces acting reciprocally on the anchorage unit result in significant anchorage loss in the palatally-inserted titanium miniscrews used for added anchorage support: we observed ventral movement in the vicinity of the miniscrew heads of 0.95+/-0.82 mm (the mean; p = 0.005), and extrusion of 0.21+/-0.28 mm (p = 0.040). In the process they tipped 2.65 degrees +/-6.23 degrees in relation to the palatal plane and 2.15 degrees +/-5.76 degrees in relation to the anterior cranial base. We observed no evidence of direct screw-to-bone contact in any of the explanted miniscrews. CONCLUSIONS: Titanium miniscrews with a polished surface, 1.6 mm in diameter and 8-9 mm long, do not provide stationary anchorage in molar distalization with the periodontally/miniscrew-anchored Distal Jet. When subjected for several months to load from forces that act in reciprocity to the force systems occurring during molar distalization, they fail to remain completely stationary in position in the palatal locations in which they were inserted. However, the combined anchorage setup is sufficient, intraorally and regardless of patient compliance, to largely compensate for the mesially-acting forces that occur reciprocal to molar distalization. PMID- 18385958 TI - Microdamage in cortical bone due to the overtightening of orthodontic microscrews. AB - BACKGROUND AND AIM: The possibility of skeletal anchorage achieved with mini implants has greatly broadened the spectrum of treatment modalities in orthodontics. Aim of this study was to investigate the microstructural alterations in cortical bone due to overtightening of orthodontic microscrews during the insertion procedure. MATERIAL AND METHODS: After having prepared three fresh pelvic porcine bone segments and removed the periosteum, we drilled 25 holes 1 mm in diameter into the segments. Using a screwdriver, we then manually inserted a total of 20 orthodontic microscrews from the Aarhus Anchorage System (1.5 mm x 7.7 mm) into the holes: ten screws were inserted about 6.7 mm deep, leaving the last 1 mm of the thread visible outside the bone. The other ten screws were initially inserted 7.7 mm (until the screw thread was no longer visible and the screw neck was slightly touching the cortex). Those last ten screws were then tightened by another quarter-turn. Five drill-holes were left empty, serving as a reference. The three pelvic bone segments were sawed into 25 smaller bone-specimens, 20 segments containing one screw and five with only the drill-hole. They were cortically cross-sectioned and processed for scanning electron microscope analysis. All the microcracks in the peri-implant bone tissue at least 50 microm long were documented for each sample. Statistical analysis was carried out according to the "number of cracks", "accumulated length of all cracks", "maximum radius of crack alteration", and "longest crack". The specimens were then divided into two subgroups according to cortical thickness. RESULTS: The group of overtightened implants showed higher values in the following categories: number of cracks, accumulated length of all cracks, maximum radius of crack alteration, and longest crack in comparison to the reference samples without screws and the group of less deeply-inserted implants. However, we identified no correlation between cortical thickness and the degree of microdamage. CONCLUSIONS: We demonstrate that there was more microstructural damage in cortical bone due to overtightening through deep insertion of orthodontic microscrews than occurred at the lower insertion depth. Extensive osseous microdamage may detract from the stability of immediately-loaded microscrews and implants due to the bone-remodeling processes initiated by microdamage. The subject of just how serious this potential risk is should be addressed in future studies. PMID- 18385959 TI - Lingual orthodontics with orthognathic surgery in a severe class II, division 2 case. AB - OBJECTIVE: To what extent are incisor protrusion, intrusion and torque feasible with orthodontic therapy in an adult? Does a lingually-fixed appliance lend itself to the controlled execution of such extensive tooth movements, and can such an appliance be used in an orthognathic surgical approach with mandibular repositioning? PATIENT AND METHODS: The treatment of a 47-year-old male patient presenting a severe Angle Class II, Division 2 with traumatic deep overbite is demonstrated in all phases of an orthodontic-surgical therapeutic approach. The preoperative harmonization of the upper and lower dental arches preceded the surgical correction via a bilateral sagittal split osteotomy in the gonial angle according to Hunsuck-Epker. Postoperative therapy included compensation for the surgical overcorrection, and the final setting of the occlusion. RESULTS: Extensive tooth movements of the incisors in particular were realized with a lingually-fixed appliance. The appliance did not hamper the surgical correction in any way. Key changes in several cephalometric values were observed: the U1:NL(ANS-PNS) angle (70 degrees) changed from 106 degrees to 70 degrees, the U1:N-S (77 degrees) from 110 degrees to 76 degrees, the distance U1: NA (4 mm) from -8 mm to +3 mm, and the interincisal angle (135 degrees) from 183 degrees to 134 degrees (average values in parentheses). Standard clinical values were obtained for the upper and lower incisors, and complete leveling of the arches was achieved. The orthognathic surgical intervention changed the Wits value from +7 to 0 mm. The intended result (as laid out in the set-up) was realized. CONCLUSION: This case report demonstrates that extensive and difficult orthodontic treatment requiring orthognathic surgery to correct a skeletal malocclusion is feasible with a fixed lingual appliance. PMID- 18385960 TI - [Depression and suicide]. AB - Depression is the most common psychiatric disorder in the general community. Depression is accompanied with the highest suicide risk. 3 % to 4 % of all depressives die by suicide even today. Suicide prevention is an important part of diagnostics, therapy, the long-term treatment, and care of depressed patients. Psychotherapeutic and psychopharmacological aspects must be included. PMID- 18385961 TI - [Optimised services for depressed patients. The German Alliance Against Depression]. AB - Optimised treatment of depression is important to patients and their families and has important health economic implications in Germany. The optimised treatment of depression can best be achieved with multilevel interventions targeting different groups. The "German Alliance Against Depression" emerged within the framework of the "German Research Network on Depression and Suicidality". This action-oriented programme aims to improve the care of depressed patients through parallel interventions on four levels. These levels are as follows: 1) co-operation with general practitioners, 2) a public relations campaign, 3) training programmes for community facilitators and 4) interventions with high-risk groups. The programme was tested in Nuremberg. Results were promising with a 24% decrease in suicidal behaviour reported. In the meantime, groups in Germany and abroad declared their interest in adapting the programme. Today, close to 40 German regions have implemented similar multilevel interventions. The European Alliance Against Depression (EAAD) formed and now includes 17 countries. The EAAD was mentioned in the 2005 EU green paper on mental health as an "example for best practice". Improving the care of affected persons requires a systematic research approach. The "Mental Health Research Network" contributes to research related to the optimisation of services for persons with mental disorders. PMID- 18385962 TI - [Depression as a comorbid disorder in primary care]. AB - As part of the DETECT study, a nationwide representative clinical-epidemiological study, the frequency and associated problems of comorbid depression with a wide range of somatic illnesses were studied in N = 51,000 primary care patients. Further the association with health related quality of life and disability is examined. Depression was assessed with the Depression Screening Questionnaire (DSQ) with an ICD-10 algorithm. RESULTS: (1) 7.5 % of all primary care patients met criteria for ICD-10 depressive disorders. (2) Depression risk was increased whenever any somatic disorder was present and increased in a dose-response relationship by number of comorbid conditions. (3) Elevation of depression risk was fairly independent of type of diagnosis, although associations with coronary heart disease (OR: 1.7), diabetic complications (OR: 1.7- 2.0), stroke (OR: 2.5) and pain-related chronic disorders (OR: 1.5) were particularly pronounced. Moderate associations were found for hyperlipidaemia (OR: 1.1). (4) Associated with the increasing number of comorbid conditions, patients with comorbid depression had increasingly more disability days and lower health related quality of life. It is concluded that the degree to which the frequency and the deleterious effects of comorbid depression is underestimated and unrecognized is alarming. The use of comorbidity indices might improve recognition. PMID- 18385963 TI - [Depressive disorders in children and adolescents]. AB - Depressive disorders in children and adolescents are widespread, cause a strong impairment in everyday life and are of great risk for further development. This paper gives an overview on the symptomatology, epidemiology, comorbid disorders and the further course of the disorder. Etiologic factors are presented and empiric studies concerning the treatment of depressive disorders in children and adolescents are summarized. Based on the empirical evidence and clinical experience suggestions for the treatment are provided. PMID- 18385964 TI - [Diagnostics of depressive disorders]. AB - Many reliable and valid instruments for screening, assessment of severity of depressive disorders and classification according to ICD-10 or DSM-IV criteria are available. Most instruments are available as self-, as well as observer-based rating questionnaires and interviews. The aim of this overview is to give a comprehensive review of well-established instruments in German speaking countries. PMID- 18385965 TI - Injury risk in young psychiatric outpatients: an 11-year follow-up of 302 adolescents. AB - BACKGROUND: Studies investigating the association between injuries and mental health have mainly focused on mental health sequelae of injuries. The aim of this prospective cohort study was to assess the incidence and risk factors of physical injury hospitalisation and poisoning hospitalisation among adolescent psychiatric outpatients. SUBJECTS AND METHODS: Data on 302 consecutively referred Finnish psychiatric outpatients aged 12-22 years (mean 16) were collected at treatment entry. The end-point of the average 11-year follow-up was death or end of follow up on 31 December 2005. The main outcome variables were physical injury hospitalisation and poisoning hospitalisation. RESULTS: Altogether 111 physical injury hospitalisations occurred in 65 (22% of all) persons during follow-up, incidence being 27.9 (95% CI: 22.7-33.1) per 1,000 person-years. Poisoning hospitalisation occurred in 22 (7.3%) persons, altogether 50 times, incidence being 12.6 (95% CI: 9.1-16.0). Seven injury-related deaths occurred, incidence being 1.8 (95% CI: 0.5-3.1) per 1,000 person-years. The most common physical injury types were fractures (40%), followed by distortions (10%) and wounds (10%), while poisoning for drugs accounted for 72% of the poisonings. Previous inpatient care, psychotropic medication, suicidality, and major depression were associated with poisoning hospitalisation during the follow-up while only gender was associated with physical injury hospitalisation. CONCLUSION: Injuries cause significant morbidity among psychiatric outpatients, but only poisonings seem to be related with suicidality in Finnish adolescent psychiatric outpatients. The high frequency of injuries seems to justify clinicians' attention to these aspects when assessing the need for care among young people. PMID- 18385966 TI - A prevalence study of current tobacco smoking in later life community and its association with sociodemographic factors, physical health and mental health status. AB - OBJECTIVE: Evaluate the frequency of current smoking in elderly people living in urban areas of Rio Grande do Sul, Brazil. METHODS: Cross-sectional design. A representative sample of 6,961 elderly, randomly selected subjects, living in a community, was examined to estimate the frequency of current tobacco smoking. Tobacco use was measured by means of a household questionnaire administered by trained interviewers that inquired about current tobacco use, sociodemographic characteristics, self-rated physical and health status. Mental health was evaluated using the Short Psychiatric Evaluation Schedule (SPES). RESULTS: The prevalence of tobacco use was 28.9% among men, 13.6% among women and 18.8% for both sexes. Male gender (OR = 3.25), low income (OR = 1.52), years of schooling (illiterate) (OR = 1.35), non-Protestant religion (OR = 2.17) and absence of physical exercise (OR = 1.21) presented positive and independent association with tobacco use. Presence of pulmonary disease (OR = 1.93) and mental distress (OR = 1.32) and absence of cardiac disease (OR = 1.51), high blood pressure (OR = 1.51) and diabetes (OR = 1.50) were independently associated with an increased chance of current tobacco use. Increasing age (OR = 0.93) and marital status (married) (OR = 0.66) presented independent and negative association with smoking. CONCLUSION: Factors associated with an increased chance of tobacco smoking were: being men, illiterate, with lower income, presence of respiratory and mental disease, and absence of cardiac disease, high blood pressure and diabetes. Factors associated with a decreased risk of tobacco smoking were: aging, exercise, Protestant religion and marriage. PMID- 18385967 TI - Chemokine-like functions of MIF in atherosclerosis. AB - The cytokine macrophage migration inhibitory factor (MIF) is a unique pro inflammatory regulator of many acute and chronic inflammatory diseases. In the pathogenesis of atherosclerosis, chronic inflammation of the arterial wall characterized by chemokine-mediated influx of leukocytes plays a central role. The contribution of MIF to atherosclerotic vascular disease has come into focus of many studies in recent years. MIF is highly expressed in macrophages and endothelial cells of different types of atherosclerotic plaques, and functional studies established the contribution of MIF to lesion progression and plaque inflammation. This proatherogenic effect may partly be explained by the finding that MIF regulates inflammatory cell recruitment to lesion areas. Similar to chemokines, MIF induces integrin-dependent arrest and transmigration of monocytes and T cells. These chemokine-like functions are mediated through interaction of MIF with the chemokine receptors CXCR2 and CXCR4 as a non-canonical ligand. In atherogenic monocyte recruitment, MIF-induced monocyte adhesion involves CD74 and CXCR2, which form a signaling receptor complex. In addition to lesion progression, MIF has been implicated in plaque destabilization, since MIF is predominantly expressed in vulnerable plaques and can induce collagen-degrading matrix metalloproteinases. The latter could be a relevant mechanism in atherosclerotic abdominal aneurysm formation, where MIF expression is correlated with aneurysmal expansion. In summary, MIF has been identified as an important regulator of atherosclerotic vascular disease with exceptional chemokine-like functions. Detailed analysis of the interaction of MIF with its receptors could provide valuable information for drug development for the anti-inflammatory treatment of established and unstable atherosclerosis. PMID- 18385969 TI - Curbing the appetites of the big eaters. PMID- 18385968 TI - Brain renin angiotensin in disease. AB - A brain renin angiotensin system (RAS) and its role in cardiovascular control and fluid homeostasis was at first controversial. This was because a circulating kidney-derived renin angiotensin system was so similar and well established. But, the pursuit of brain RAS has proven to be correct. In the course of accepting brain RAS, high standards of proof attracted state of the art techniques in all the new developments of biology. Consequently, brain RAS is a robust concept that has enlightened neuroscience as well as cardiovascular physiology and is a model neuropeptide system. Molecular biology confirmed the components of brain RAS and their location in the brain. Transgenic mice and rats bearing renin and extra copies of angiotensinogen genes revealed the importance of brain RAS. Cre-lox delivery in vectors has enabled pinpoint gene deletion of brain RAS in discrete brain nuclei. The new concept of brain RAS includes ACE-2, Ang1-7, and prorenin and Mas receptors. Angiotensin II (ANG II) generated in the brain by brain renin has many neural effects. It activates behavioral effects by selective activation of ANG II receptor subtypes in different locations. It regulates sympathetic activity and baroreflexes and contributes to neurogenic hypertension. New findings implicate brain RAS in a much wider range of neural effects. We review brain RAS involvement in Alzheimer's disease, stroke memory, and learning alcoholism stress depression. There is growing evidence to consider developing treatment strategies for a variety of neurological disease states based on brain RAS. PMID- 18385970 TI - [Treatment and care strategies for Alzheimer's disease and related dementias]. AB - The majority of elderly demented patients suffers from mixed dementia, with Alzheimer's-type brain changes being the leading cause of cognitive impairment. Cholinesterase inhibitors and memantin improve cognitive performance in dementia, with a symptomatic parallel shift of 8 to 12 months. Disturbances of perception and behaviour can be prohibited or mitigated by antidementia drugs and improved with antidepressants, neuroleptics, and other substances. The somatic and cerebral comorbidity of demented patients offers further options for improving sensory input, cerebral availability of oxygen and glucose, and even secondary prevention. Technical support systems for demented patients which are already available remain largely underused. Psychosocial support is mandatory, even though the advantages of specific interventions have not been demonstrated with scientific rigour. PMID- 18385971 TI - Long-term prevention of diabetic nephropathy: an audit. AB - AIMS/HYPOTHESIS: In type 1 diabetic patients with microalbuminuria not receiving antihypertensive treatment, an increase in urinary AER (UAER) of 6-14%/year and a risk of developing diabetic nephropathy (DN) of 3-30%/year have been reported. We audited the long-term effect of blocking the renin-angiotensin-aldosterone system (RAAS) with an ACE inhibitor (ACEI) or angiotensin II receptor blocker (ARB) in microalbuminuric type 1 diabetic patients on progression of microalbuminuria and development of DN. METHODS: All patients with type 1 diabetes and persistent microalbuminuria (30-300 mg/24 h) were identified (n=227) in 1995 at Steno Diabetes Center and followed for 11 years. Development of DN was defined as a UAER of >300 mg/24 h in two of three consecutive urine samples. RESULTS: Age and duration of diabetes at baseline (mean+/-SD) were 46+/-15 and 28+/-13 years, respectively. During follow-up 14 patients emigrated and 58 (26%) died. Over the same period 79% were treated with an ACEI or ARB. There was a mean decline in UAER of 4%/year. Sixty-five patients (29%) progressed to overt DN, corresponding to 3.1%/year. However, 29 of them regressed to normo- or microalbuminuria on intensified antihypertensive treatment. Glycaemic control and blood pressure remained nearly unchanged. CONCLUSIONS/INTERPRETATION: In our outpatient clinic, the implementation of RAAS-blocking treatment in type 1 diabetic patients with microalbuminuria successfully reduced long-term progression to overt DN to a rate similar to those previously reported in randomised, double-blind intervention trials of shorter duration using RAAS blockade. PMID- 18385972 TI - Discovery of dipeptidyl-peptidase IV--a 40 year journey from bench to patient. PMID- 18385973 TI - Functional significance of repressor element 1 silencing transcription factor (REST) target genes in pancreatic beta cells. AB - AIMS/HYPOTHESIS: The expression of several neuronal genes in pancreatic beta cells is due to the absence of the transcription factor repressor element 1 (RE 1) silencing transcription factor (REST). The identification of these traits and their functional significance in beta cells has only been partly elucidated. Herein, we investigated the biological consequences of a repression of REST target genes by expressing REST in beta cells. METHODS: The effect of REST expression on glucose homeostasis, insulin content and release, and beta cell mass was analysed in transgenic mice selectively expressing REST in beta cells. Relevant target genes were identified in INS-1E and primary beta cells expressing REST. RESULTS: Transgenic mice featuring a beta cell-targeted expression of REST exhibited glucose intolerance and reduced beta cell mass. In primary beta cells, REST repressed several proteins of the exocytotic machinery, including synaptosomal-associated protein (SNAP) 25, synaptotagmin (SYT) IV, SYT VII, SYT IX and complexin II; it impaired first and second phases of insulin secretion. Using RNA interference in INS-1E cells, we showed that SYT IV and SYT VII were implicated in the control of insulin release. CONCLUSIONS/INTERPRETATION: The data document the critical role of REST target genes in pancreatic beta cells. Specifically, we provide evidence that the downregulation of these genes is detrimental for the exocytosis of large dense core vesicles, thus contributing to beta cell dysfunction and impaired glucose homeostasis. PMID- 18385975 TI - Primarily non-surgical management of osteomyelitis of the foot in diabetes. AB - AIMS/HYPOTHESIS: We examined the use of surgery and assessed the response to non surgical management of osteomyelitis of the foot in diabetic patients. METHODS: We reviewed the records of all patients presenting to a single specialist centre with osteomyelitis complicating a diabetic foot ulcer over a 5 year period. Details were extracted on antibiotic choice and treatment duration, hospital admission, incidence of minor and major amputation, and 12 month outcomes. RESULTS: There were 147 patients, with mean age 64.7 years (66% men). Of these, 26 (18%) were admitted to hospital at the time of presentation and managed with intravenous antibiotics; the remainder were managed with oral antibiotics as outpatients. Surgery was undertaken because of life- or limb-threatening infection, or failure to respond, in 34 (23%) patients (minor amputation 28, major amputation six patients). The remaining 113 were managed non-surgically. Remission was induced in 66 (58.4% of 113), while 35 (31%) had a relapse. Of those experiencing relapse, 27 (77%) achieved apparent arrest of the infection with a further course of antibiotics; six underwent minor and two underwent major amputation. Of all 113 whose infection was initially managed without surgery, apparent remission was achieved with antibiotics alone in 93 (82.3%). CONCLUSIONS/INTERPRETATION: As these observations were made in an unselected case series, they give more insight into the respective roles of surgical and non surgical management. The results confirm that although urgent surgery is indicated in some patients, non-surgical management of those without limb threatening infection is associated with a high rate of apparent remission. PMID- 18385976 TI - [Concepts of in-patient gradual diagnostics for patients with lumbar back-pain]. AB - Concepts for treating back pain by considering recent advancements in understanding chronic pain have been increasingly discussed over the last years. In general, any kind of therapy requires a most accurate diagnosis; however, in cases of lumbar back pain matching clinical observations with known structural pathologies is not always straight forward. Here, we suggest a concept of in patient gradual diagnosis of patients with back pain that includes stepwise structural and functional components. We emphasize that in addition to the specific (classifiable) factors causing back pain (e.g. compression of lumbar nerve roots, facet joint associated pain syndrome, sacroiliacal dysfunction, segmental instability) also unspecific (non-classifiable) causes as well as disease-associated and/or determining bio-psychological or social factors need to be considered. With this study, we aim to introduce the procedure and assess the value of in-patient gradual diagnostics. We will describe different approaches, which we will evaluate for specific indications. PMID- 18385977 TI - Development and simultaneous application of multiple care protocols in critical care: a multicenter feasibility study. AB - OBJECTIVE: To test the feasibility of and interactions among three software driven critical care protocols. DESIGN: Prospective cohort study. SETTING: Intensive care units in six European and American university hospitals. PATIENTS: 174 cardiac surgery and 41 septic patients. INTERVENTIONS: Application of software-driven protocols for cardiovascular management, sedation, and weaning during the first 7 days of intensive care. MEASUREMENTS AND RESULTS: All protocols were used simultaneously in 85% of the cardiac surgery and 44% of the septic patients, and any one of the protocols was used for 73 and 44% of study duration, respectively. Protocol use was discontinued in 12% of patients by the treating clinician and in 6% for technical/administrative reasons. The number of protocol steps per unit of time was similar in the two diagnostic groups (n.s. for all protocols). Initial hemodynamic stability (a protocol target) was achieved in 26+/-18 min (mean+/-SD) in cardiac surgery and in 24+/-18 min in septic patients. Sedation targets were reached in 2.4+/-0.2h in cardiac surgery and in 3.6 +/-0.2h in septic patients. Weaning protocol was started in 164 (94%; 154 extubated) cardiac surgery and in 25 (60%; 9 extubated) septic patients. The median (interquartile range) time from starting weaning to extubation (a protocol target) was 89 min (range 44-154 min) for the cardiac surgery patients and 96 min (range 56-205 min) for the septic patients. CONCLUSIONS: Multiple software-driven treatment protocols can be simultaneously applied with high acceptance and rapid achievement of primary treatment goals. Time to reach these primary goals may provide a performance indicator. PMID- 18385978 TI - Improved short-term survival with polyethylene glycol modified hemoglobin liposomes in critical normovolemic anemia. AB - OBJECTIVE: To investigate the efficacy of a polyethylene glycol (PEG) modified formulation of liposome-encapsulated hemoglobin (LEH) as an oxygen-carrying blood substitute in the treatment of critical normovolemic anemia. DESIGN AND SETTING: Prospective, controlled, randomized experimental study in a university research facility. SUBJECTS: 14 anesthetized and mechanically ventilated beagle dogs. INTERVENTIONS: Animals were splenectomized and hemodiluted by exchange of whole blood for iso-oncotic hetastarch (HES). Target parameter of the hemodilution protocol was the individual critical hemoglobin concentration (Hb(crit)) corresponding with the onset of O(2) supply dependency of total body O(2) consumption. At Hb(crit) animals were randomized to receive a bolus infusion (20[Symbol: see text]ml/kg) of either LEH (n = 7) or normal saline (NS; n = 7). Subsequently animals were observed without further intervention. MEASUREMENTS AND RESULTS: The primary endpoint was survival time after the completion of treatment; secondary endpoints were parameters of central hemodynamics, O(2) transport and tissue oxygenation. Animals in the LEH group survived significantly longer after completion of treatment (149 +/- 109 vs. 43+/- 56 min). Immediately after treatment LEH-treated animals presented with a more stable cardiovascular condition. After 30 min tissue O(2) tension on the surface of a skeletal muscle was significantly higher in the LEH group (23+/-8 vs. 9 +/- 2 mmHg). Nevertheless, treatment with LEH did not decrease mortality within the observation period. CONCLUSIONS: In this present experimental study the infusion of a PEG-modified LEH provided adequate tissue oxygenation, hemodynamic stability, and a prolongation of survival time after critical anemia. However, these effects were sustained for only a short period of time. PMID- 18385979 TI - Analysis and prevention of intra-operative complications of TransFix fixation in anterior cruciate ligament reconstruction. AB - We encountered some complications during the graft passage and fixation using a TransFix implant. Despite the large number of cases and publications related to the ligament surgery, few papers have reported intra-operative complications. We report our experience of such complications and prevention methods. From June 2005 to October 2007, anterior cruciate ligament reconstructions were performed in 128 patients using the TransFix devices. From the surgical records, the opinions of the assistant during the graft passage were divided into three subsets: tight, modest, and loose. The results were also analyzed according to the use of the routine procedure, vertical traction, and thigh portal. Complications during the graft passage and fixation were encountered in 13 patients. These complications occurred when the femoral tunnels were tight during the graft passage (P < 0.0001). The addition of vertical traction or a thigh portal decreased the number of complications significantly (P < 0.0001). The main cause of such complications was the lack of full proximal migration of the graft, which can cause tendon and wire breakage. This study introduces some methods to prevent such complications. PMID- 18385981 TI - Ladderane lipid distribution in four genera of anammox bacteria. AB - Intact ladderane phospholipids and core lipids were studied in four species of anaerobic ammonium oxidizing (anammox) bacteria, each representing one of the four known genera. Each species of anammox bacteria contained C18 and C20 ladderane fatty acids with either 3 or 5 linearly condensed cyclobutane rings and a ladderane monoether containing a C20 alkyl moiety with 3 cyclobutane rings. The presence of ladderane lipids in all four anammox species is consistent with their putative physiological role to provide a dense membrane around the anammoxosome, the postulated site of anammox catabolism. In contrast to the core lipids, large variations were observed in the distribution of ladderane phospholipids, i.e. different combinations of hydrophobic tail (ladderane, straight chain and methyl branched fatty acid) types attached to the glycerol backbone sn-1 position, in combination with different types of polar headgroup (phosphocholine, phosphoethanolamine or phosphoglycerol) attached to the sn-3 position. Intact ladderane lipids made up a high percentage of the lipid content in the cells of "Candidatus Kuenenia stuttgartiensis", suggesting that ladderane lipids are also present in membranes other than the anammoxosome. Finally, all four investigated species contained a C27 hopanoid ketone and bacteriohopanetetrol, which, indicates that hopanoids are anaerobically synthesised by anammox bacteria. PMID- 18385980 TI - Effects of biophysical stimulation in patients undergoing arthroscopic reconstruction of anterior cruciate ligament: prospective, randomized and double blind study. AB - Pre-clinical studies have shown that treatment by pulsed electromagnetic fields (PEMFs) can limit the catabolic effects of pro-inflammatory cytokines on articular cartilage and favour the anabolic activity of the chondrocytes. Anterior cruciate ligament (ACL) reconstruction is usually performed by arthroscopic procedure that, even if minimally invasive, may elicit an inflammatory joint reaction detrimental to articular cartilage. In this study the effect of I-ONE PEMFs treatment in patients undergoing ACL reconstruction was investigated. The study end-points were (1) evaluation of patients' functional recovery by International Knee Documentation Committee (IKDC) Form; (2) use of non-steroidal anti-inflammatory drugs (NSAIDs), necessary to control joint pain and inflammation. The study design was prospective, randomized and double blind. Sixty-nine patients were included in the study at baseline. Follow-up visits were scheduled at 30, 60 and 180 days, followed by 2-year follow-up interview. Patients were evaluated by IKDC Form and were asked to report on the use of NSAIDs. Patients were randomized to active or placebo treatments; active device generated a magnetic field of 1.5 mT at 75 Hz. Patients were instructed to use the stimulator (I-ONE) for 4 h per day for 60 days. All patients underwent ACL reconstruction with use of quadruple hamstrings semitendinosus and gracilis technique. At baseline there were no differences in the IKDC scores between the two groups. At follow-up visits the SF-36 Health Survey score showed a statistically significant faster recovery in the group of patients treated with I ONE stimulator (P < 0.05). NSAIDs use was less frequent among active patients than controls (P < 0.05). Joint swelling resolution and return to normal range of motion occurred faster in the active treated group (P < 0.05) too. The 2-year follow-up did not shown statistically significant difference between the two groups. Furthermore for longitudinal analysis the generalized linear mixed effects model was applied to calculate the group x time interaction coefficient; this interaction showed a significant difference (P < 0.0001) between the active and placebo groups for all investigated variables: SF-36 Health Survey, IKDC Subjective Knee Evaluation and VAS. Twenty-nine patients (15 in the active group; 14 in the placebo group) underwent both ACL reconstruction and meniscectomy; when they were analysed separately the differences in SF-36 Health Survey scores between the two groups were larger then what observed in the whole study group (P < 0.05). The results of this study show that patient's functional recovery occurs earlier in the active group. No side effects were observed and the treatment was well tolerated. The use of I-ONE should always be considered after ACL reconstruction, particularly in professional athletes, to shorten the recovery time, to limit joint inflammatory reaction and its catabolic effects on articular cartilage and ultimately for joint preservation. PMID- 18385982 TI - Regulation of gvp genes encoding gas vesicle proteins in halophilic Archaea. AB - Three gas vesicle gene clusters derived from Halobacterium salinarum (p-vac and c vac) and Haloferax mediterranei (mc-vac) are used as model systems to study gene regulation in Archaea. An unusual pair of regulatory proteins is involved here, with GvpE acting as transcription activator and GvpD exhibiting a repressing function. Both regulators are able to interact leading to the loss of GvpE and the repression (or turnoff) of the gas vesicle formation. The latter function of GvpD requires a p-loop motif and an arginine-rich region, bR1. Both regulator proteins are differentially expressed from the same gvp transcript in Hfx. mediterranei and Hbt. salinarum PHH4. GvpE appears to recognize a 20-nucleotide activator sequence (UAS) located upstream and adjacent to the TFB-recognition element BRE of the two promoters driving the transcription of the divergently oriented gvpACNO and gvpDEFGHIJKLM gene clusters. The BRE elements of these two promoters are separated by 35 nucleotides only, and the distal portions of the two GvpE-UAS overlap considerably in the center of this region. Mutations here negatively affect the GvpE-induced activities of both gvp promoters, whereas alterations in the proximal UAS portions only affect the activity of the promoter located close by. PMID- 18385983 TI - Escherichia coli tat mutant strains are able to transport maltose in the absence of an active malE gene. AB - The twin-arginine transport (Tat) system is a prokaryotic protein transport system. Escherichia coli mutants in this pathway show a defect in cell separation during cell division, resulting in destabilization and permeability of the outer membrane. Maltose uptake is catalysed by a membrane-bound transporter of the ATP binding cassette (ABC) superfamily, where MalE is the essential periplasmic binding protein component. Here, we report that tat mutants are unexpectedly able to transport maltose in the absence of malE. This observation is specific to the MalE component since co-inactivation of malF, which encodes one of the channel components of the transporter, completely abolishes maltose transport even when the Tat system is inactivated. Genetic repair of the outer membrane leaky phenotype of the tat mutant strain re-established the absolute requirement for MalE in maltose uptake. In addition, we demonstrate that phenotypic repair of the outer membrane defect of the tat strain can also be achieved chemically by the inclusion of high concentrations of calcium or magnesium in the growth medium. PMID- 18385984 TI - Interleukin 8 and susceptibility to coronary artery disease: a population genetics perspective. AB - INTRODUCTION: Interleukin 8 is a strong chemoattractant factor for neutrophils and T lymphocytes. We investigated the potential influence of two common polymorphisms of the interleukin-8 gene, -251A/T, and 781C/T on susceptibility to coronary artery disease. MATERIALS AND METHODS: The hypothesis was tested by screening for the prevalence of the above polymorphisms in 241 angiographically diagnosed coronary artery disease patients compared to 157 selected controls with negative coronary angiography. RESULTS AND DISCUSSION: We found no significant differences between cases and controls concerning the allelic and genotypic frequencies of both the studied polymorphisms. Nevertheless, a statistically significant lower frequency of the AA containing genotypes was observed in cases presenting with acute coronary syndromes compared to asymptomatic subjects or patients with stable coronary artery disease (OR = 0.44, 95%CI: 0.2-0.98, p = 0.04). The strongest statistical significance was observed in the AA(251)TT(781) combined genotype (OR = 0.34, 95%CI: 0.14-0.85, p = 0.02). CONCLUSION: Our results suggest that the genetic diversity of the interleukin-8 gene influences the clinical manifestation of CAD. PMID- 18385985 TI - Effects of Cd2+ on transient outward and delayed rectifier potassium currents in acutely isolated rat hippocampal CA1 neurons. AB - The effects of cadmium (Cd(2+)) on the transient outward potassium current (I(A)) and delayed rectifier potassium current (I(K)) were investigated in acutely dissociated rat hippocampal CA1 neurons using the whole-cell patch-clamp technique. The results showed that Cd(2+) inhibited the amplitudes of I(A) and I (K) in a reversible and concentration-dependent manner, with half-maximal inhibitive concentration (IC(50)) values of 546+/-59 and 749+/-53 microM, and the inhibitory effect of Cd(2+) was voltage dependent. Cd(2+) significantly shifted the steady-state activation and inactivation curve of I(A) to more positive potentials. In contrast, Cd(2+) caused a relatively less but still significant positive shift in the activation of I(K) without effect on the inactivation curve. Cd(2+) significantly slowed the recovery from inactivation of I(K) but had no effect on the recovery time course of I(A). The results suggest that the modulation of I(A) and I(K) was most likely mediated by the interaction of Cd(2+) with a specific site on the potassium-channel protein rather than by screening of bulk surface-negative charge. The effects of Cd(2+) on the voltage-gated potassium currents may be a possible contributing mechanism for the Cd(2+) induced neurotoxic damage. In addition, the effects of Cd(2+) on the potassium currents at concentrations that overlap with its effects on calcium currents raise concerns about its use in pharmacological or physiological studies. PMID- 18385986 TI - Interactions between age and the aversive effects of nicotine withdrawal under mecamylamine-precipitated and spontaneous conditions in male Wistar rats. AB - RATIONALE: Adolescent onset of smoking is associated with a rapid progression to dependence. Although adolescents may exhibit a greater susceptibility to nicotine addiction, relatively little is known about the influence of the aversive effects of nicotine withdrawal in maintaining smoking behavior. OBJECTIVES: The present study investigated age differences in the motivational effects of mecamylamine precipitated and spontaneous nicotine withdrawal in adolescent and adult rats using the conditioned place aversion procedure (CPA). MATERIALS AND METHODS: In experiment 1, adolescent (postnatal day (PD) 28) and adult (PD60) male Wistar rats chronically treated with nicotine (3 or 6 mg/kg/day, s.c.) received mecamylamine (1 mg/kg, s.c.), a nicotinic receptor antagonist, or vehicle prior to place conditioning; physical withdrawal signs were also measured. Experiment 2 was conducted to increase nicotine levels in which adolescents were treated with 4.5 or 9 mg/kg/day nicotine. In experiment 3, age differences in spontaneous nicotine withdrawal were evaluated. RESULTS: Nicotine-treated adults developed a CPA to the mecamylamine-associated compartment and expressed significant physical withdrawal signs, whereas similarly treated adolescents did not. Increasing nicotine exposure levels did not modify the adolescent response to mecamylamine precipitated withdrawal. Spontaneous nicotine withdrawal produced similar physical withdrawal signs in adolescents and adults, but did not elicit CPA. CONCLUSIONS: The current study indicates that adolescent rats are less responsive to the aversive effects of mecamylamine-precipitated, but not spontaneous, nicotine withdrawal compared to adult rats. These findings suggest that adolescents and adults may exhibit similar sensitivity to the affective and physical effects of withdrawal following smoking cessation. PMID- 18385987 TI - Inductively coupled plasma mass spectrometric analysis of the total amount of platinum in DNA extracts from peripheral blood mononuclear cells and tissue from patients treated with cisplatin. AB - We present a highly sensitive method for the determination of platinum (Pt) in DNA extracts of peripheral blood mononuclear cells (PBMCs) and tissue samples from patients treated with cisplatin. The method is based on the measurement of Pt by inductively coupled plasma mass spectrometry (ICP-MS) and allows quantification of Pt-DNA adducts in PBMCs isolated from 10 mL blood and 1 mg tissue. The lower limit of quantification is 0.75 pg Pt or 7.5 fg Pt MUg(-1) DNA when using 100 MUg DNA. The method proved to be accurate and precise. The results obtained using the ICP-MS method were in good agreement with results from the alternative (32)P-postlabelling assay. The ICP-MS method was, however, more sensitive and proved to be less laborious. The advantages of the presented ICP-MS technique were demonstrated by the analysis of PBMCs, normal gastric tissue, and gastric tumour tissue of patients treated with cisplatin. PMID- 18385988 TI - High- and low-frequency repetitive transcranial magnetic stimulation differentially activates c-Fos and zif268 protein expression in the rat brain. AB - Repetitive transcranial magnetic stimulation (rTMS) has been shown to alter cortical excitability depending on the stimulus-frequency used, with high frequency (5 Hz and higher) increasing it but low frequency (usually 1 Hz or lower) reducing it. To determine the efficiency of different rTMS protocols in inducing cortical network activity, we tested the acute effect of one low frequency rTMS protocol (1 Hz) and two different high-frequency protocols (10 Hz and intermittent theta-burst stimulation, iTBS) on the expression of the two immediate early gene (IEG) proteins c-Fos and zif268 in the rat brain. The cortical expression of both IEGs was specifically changed in an rTMS-dependent manner. One and 10 Hz rTMS enhanced c-Fos protein expression in all cortical areas tested, while iTBS was effective only in limbic cortices. Zif268 expression was increased in almost all cortical areas after iTBS, while 10 Hz rTMS was effective only in the primary motor and sensory cortices. One Hertz rTMS had no effect on cortical zif268 expression. Furthermore, sham-rTMS had no effect on zif268 expression but increased c-Fos in limbic cortices. This is the first study demonstrating that cortical zif268 and c-Fos expression can be specifically modulated by acute rTMS depending on the pattern of stimulation applied. PMID- 18385989 TI - Functional neuroanatomy of the human premotor oculomotor brainstem nuclei: insights from postmortem and advanced in vivo imaging studies. AB - Considerable progress has been made recently in the field of the functional neuroanatomy of the primate oculomotor system, which has also improved our understanding of the structure, organization and function of the human oculomotor system. In the present review we provide for the first time an overview of the neuroanatomical basis of eye movement control in humans as revealed by a series of post-mortem studies in which the human premotor oculomotor brainstem nuclei were identified using unconventional 100 MUm thick serial tissue sections stained for Nissl substance and lipofuscin pigment (Nissl-pigment stain according to Braak). Data from control brains and from patients suffering from spinocerebellar ataxia type 3, a neurodegenerative disease that severely impairs oculomotor function are discussed and recommendations for the identification of human premotor oculomotor brainstem nuclei in post-mortem studies are given. To visualize premotor brainstem nuclei in living patients, modern brain imaging techniques have been employed, albeit with limited success. Establishing topographic markers of brainstem nuclei may be a necessary next step to further elucidate the functional neuroanatomy of the premotor oculomotor brainstem network in human patients. This will help radiologists to identify these nuclei in living patients and will enable clinicians to monitor the progression of neurological disorders affecting the oculomotor system. PMID- 18385990 TI - Factors predicting hospital readmissions related to adverse drug reactions. AB - OBJECTIVE: To analyse the contribution of adverse drug reactions (ADR) to hospital readmissions. METHODS: This was a case-control study in which unscheduled admissions of patients who had been admitted to the hospital during the two previous months were assessed during a 21-month period. The patient was considered a case when the main diagnosis of readmission complied with the World Health Organisation's definition of an ADR. For each case, two controls were selected from those patients that had been admitted for ADR without readmission (n = 177). Information on drugs and other risk factors was obtained from cases by interview and from controls by clinical record review. RESULTS: There were 26,559 unscheduled admissions of which 81 were readmissions associated with ADR (4.5% of the unscheduled readmissions). There were no statistically significant correlations with sex, age or medical history, with the exception of arterial hypertension. The main drug products causing readmission were acenocoumarol (15, 18.5%), antihypertensive-diuretics (14, 17.3%), anticancer drugs (11, 13.6%) and digoxin (seven, 8.6%). In the multivariate logistic analysis, the variables predicting readmission were acenocoumarol [odds ratio (OR) 12.2, 95% confidence interval (CI) 3.8-38.3, P < 0.0001], a record of diabetes mellitus (OR 2.6, 95% CI 1.3-5.5, P < 0.01), the number of drugs taken at the moment of ADR (OR 1.2, 95% CI 1.1-1.4, P < 0.001) and high blood pressure (OR 0.3, 95% CI 0.2-0.6, P < 0.001) even though the latter was a negative predictor, preventing readmission. Of the 81 readmissions associated with ADR, 28 (34.6%) were preventable. CONCLUSION: A medical record of diabetes mellitus, polypharmacy and acenocoumarol treatment were risk factors predicting hospital readmission related to ADR. PMID- 18385991 TI - Effect of CYP3A5*3 genotype on the pharmacokinetics and antiplatelet effect of clopidogrel in healthy subjects. AB - OBJECTIVE: Clopidogrel is activated by cytochrome P450 3A (CYP3A) to generate an active metabolite that inhibits adenosine diphosphate (ADP)-induced platelet aggregation through irreversible binding to the platelet P2Y12 receptor. The objective of this study was to assess the effect of the CYP3A5 genotype on the pharmacokinetics and antiplatelet effect of clopidogrel in healthy subjects. METHODS: Twenty-two healthy subjects (CYP3A5*1/*1, n = 6; CYP3A5*1/*3, n = 8; CYP3A5*3/*3, n = 8) were recruited. After the administration of a loading dose of 300 mg of clopidogrel followed by 75 mg once daily for 6 days, plasma concentrations of clopidogrel and SR26334, an inactive metabolite, were measured for 24 h. The antiplatelet effect of clopidogrel was also measured, by determining the inhibition of ADP-induced platelet aggregation for 168 h, according to CYP3A5 genotype. RESULTS: Mean plasma concentration profiles of clopidogrel and SR26334 were comparable between CYP3A5 genotype groups. In addition, the CYP3A5 genotype did not affect the pharmacokinetics of either clopidogrel or SR26334. CYP3A5 genotype also did not modulate the inhibitory effect of clopidogrel on platelet aggregation. CONCLUSION: The CYP3A5*3 genotype plays a minor role in causing interindividual variability of the disposition of clopidogrel and its antiplatelet effect in humans. PMID- 18385992 TI - Is routine ureteral stenting necessary after uncomplicated ureteroscopic lithotripsy for lower ureteral stones larger than 1 cm? AB - We evaluated the need for routine ureteral stenting after uncomplicated ureteroscopic lithotripsy (URSL) without dilation for lower ureteral stones larger than 1 cm. A total of 43 patients underwent URSL for lower ureteral stones larger than 10 mm. They were randomized into a stented (21) or an unstented (22) group. URSL was performed by using a semirigid ureteroscope and pneumatic lithotripter without ureteral dilation. Additional forceps application (AFA) was used to remove fragments > or =4 mm. Patients in each group were assessed for stone-free rate, stone size, operative time, AFA, hospitalization time, postoperative pain, irritative voiding symptoms, hematuria, re-hospitalization and stricture formation. The stone-free rate was 100% in each group. There were no statistical differences in the two groups regarding stone size, operative time, AFA, postoperative pain, hematuria and hospitalization time. However, irritative voiding symptoms of the stented group were significantly higher than those in the unstented group (P < 0.05). One patient (4.5%) in the unstented group required re-hospitalization for severe flank pain with fever (>38 degrees C) compared to one patient (4.7%) in the stented group for proximal stent migration (P > 0.05). Stricture formation was not demonstrated in either group at 3 months follow-up excretory urography (EXU). Our results demonstrate that ureteral stenting after uncomplicated URSL without dilation for lower ureteral stones larger than 1 cm does not appear to be necessary if AFA is used to remove fragments > or =4 mm, thereby reducing morbidity of patients and risk of re hospitalization. PMID- 18385995 TI - An overview of mannan structure and mannan-degrading enzyme systems. AB - Hemicellulose is a complex group of heterogeneous polymers and represents one of the major sources of renewable organic matter. Mannan is one of the major constituent groups of hemicellulose in the wall of higher plants. It comprises linear or branched polymers derived from sugars such as D-mannose, D-galactose, and D-glucose. The principal component of softwood hemicellulose is glucomannan. Structural studies revealed that the galactosyl side chain hydrogen interacts to the mannan backbone intramolecularly and provides structural stability. Differences in the distribution of D-galactosyl units along the mannan structure are found in galactomannans from different sources. Acetyl groups were identified and distributed irregularly in glucomannan. Some of the mannosyl units of galactoglucomannan are partially substituted by O-acetyl groups. Some unusual structures are found in the mannan family from seaweed, showing a complex system of sulfated structure. Endohydrolases and exohydrolases are involved in the breakdown of the mannan backbone to oligosaccharides or fermentable sugars. The main-chain mannan-degrading enzymes include beta-mannanase, beta-glucosidase, and beta-mannosidase. Additional enzymes such as acetyl mannan esterase and alpha galactosidase are required to remove side-chain substituents that are attached at various points on mannan, creating more sites for subsequent enzymatic hydrolysis. Mannan-degrading enzymes have found applications in the pharmaceutical, food, feed, and pulp and paper industries. This review reports the structure of mannans and some biochemical properties and applications of mannan-degrading enzymes. PMID- 18385993 TI - Acridine derivatives inhibit lysozyme aggregation. AB - We have screened a library of structurally distinct acridine derivatives (19 compounds) for their ability to inhibit lysozyme amyloid aggregation in vitro. Studied acridines were divided into three structurally different groups depending on the molecule planarity and type of the side chain-planar acridines, spiroacridines and tetrahydroacridines. Thioflavine T fluorescence assay and transmission electron microscopy were used for monitoring the inhibiting activity of acridines. We have found that both the structure of the acridine side chains and molecule planarity influence their antiamyloidogenic activity. The planar acridines inhibited lysozyme aggregation effectively. Spiroacridines and tetrahydroacridines had no significant effect on the prevention of lysozyme fibrillization, probably resulting from the presence of the heterocyclic 5 membered ring and non-planarity of molecule. Moreover, in the presence of some tetrahydroacridines the enhanced extent of aggregation was detected. We identified the most active acridine derivates from studied compound library characterized by low micromolar IC50 values, which indicate their possible application for therapeutic purpose. PMID- 18385994 TI - Application of biocathode in microbial fuel cells: cell performance and microbial community. AB - Instead of the utilization of artificial redox mediators or other catalysts, a biocathode has been applied in a two-chamber microbial fuel cell in this study, and the cell performance and microbial community were analyzed. After a 2-month startup, the microorganisms of each compartment in microbial fuel cell were well developed, and the output of microbial fuel cell increased and became stable gradually, in terms of electricity generation. At 20 ml/min flow rate of the cathodic influent, the maximum power density reached 19.53 W/m3, while the corresponding current and cell voltage were 15.36 mA and 223 mV at an external resistor of 14.9 Omega, respectively. With the development of microorganisms in both compartments, the internal resistance decreased from initial 40.2 to 14.0 Omega, too. Microbial community analysis demonstrated that five major groups of the clones were categorized among those 26 clone types derived from the cathode microorganisms. Betaproteobacteria was the most abundant division with 50.0% (37 of 74) of the sequenced clones in the cathode compartment, followed by 21.6% (16 of 74) Bacteroidetes, 9.5% (7 of 74) Alphaproteobacteria, 8.1% (6 of 74) Chlorobi, 4.1% (3 of 74) Deltaproteobacteria, 4.1% (3 of 74) Actinobacteria, and 2.6% (2 of 74) Gammaproteobacteria. PMID- 18385996 TI - Early prediction of response to therapy: the clinical implications in Hodgkin's and non-Hodgkin's lymphoma. PMID- 18385997 TI - Advances in prostate cancer imaging techniques and strategies. PMID- 18385998 TI - The cost effectiveness of 123I-FP-CIT SPECT imaging in patients with an uncertain clinical diagnosis of parkinsonism. AB - PURPOSE: (123)I-N-omega-fluoropropyl-2-beta-carboxymethoxy-3beta-(4 iodophenyl)nortropane ((123)I-FP-CIT) Single-photon emission computed tomography (SPECT) has been suggested to be a useful diagnostic adjunct in patients with clinically uncertain parkinsonism. We developed a pharmaco-economic (PE) model, evaluating the cost effectiveness of adding (123)I-FP-CIT SPECT to the diagnostic workup. As the model was developed before application of the diagnostic technique in real practice, a predictive validity assessment was performed based on data from a large nationwide patient registry in these patients. METHODS: A PE model, using a Markov state transition model, was created, based on literature-derived and clinical expert panel data. Effects were expressed as adequately treated years (ATY). Key input data were compared to the real-life patterns in a nationwide multi-centre clinical setting, based on a complete national registry of 1,701 consecutive patients. The change in initial diagnosis and alteration of management of the patient after SPECT were registered. RESULTS: In the PE model, it was calculated that management would change in 48.5% of patients by SPECT and that, over a 5-year period, 1.2 ATYs could be gained at a yearly additional cost of 72 euro. From the studied 1,701 patients, nigrostriatal degeneration was observed in 59.8%, the initial diagnosis was changed in 51.5%, management was altered in 49%, and cost effectiveness was increased to 358 euro per ATY. CONCLUSION: Good correspondence between assumed and observed changes in patient management was found, indicating that (123)I-FP-CIT SPECT is influential in diagnosis and management of patients with uncertain clinical diagnosis of parkinsonism. This can be achieved at a marginal added cost to the health insurance and leads to a significant gain in ATY. PMID- 18385999 TI - Length and cost of hospital stay of radioiodine ablation in thyroid cancer patients: comparison between preparation with thyroid hormone withdrawal and thyrogen. AB - PURPOSE: Treatment of thyroid cancer consists of thyroidectomy and radioiodine ablation following thyroid-stimulating hormone (TSH) stimulation. Similar ablation rates were obtained with either thyroid hormone withdrawal (THW) or rhTSH. But with rhTSH, the elimination of radioiodine is more rapid, thus reducing its whole-body retention and potentially resulting in a shorter hospital stay. The aim of this study was to assess the financial impact of a reduced length of hospital stay with the use of rhTSH. METHODS: This was a case-control study of thyroid cancer patients treated postoperatively with 3,700 MBq (100 mCi) radioiodine; 35 patients who received rhTSH were matched with 64 patients submitted to THW according to covariates influencing radioiodine retention. The length of hospitalization (LOH) was estimated for each method according to the threshold of radioiodine retention below which the patient can be discharged from the hospital. The economic analysis was conducted from a hospital perspective. Simulations were performed. RESULTS: For a threshold of 400 MBq, the LOH was 2.4 days and 3.5 days with rhTSH and THW, respectively, and the cost for an ablation stay was, respectively, 2,146 and 1,807 . In the French context, 57% of the acquisition cost of rhTSH was compensated by the reduction of the length of hospitalization. CONCLUSION: By increasing the iodine excretion, rhTSH allows a shorter hospitalization length, which partially compensates its acquisition cost. PMID- 18386000 TI - Induction of T cell-mediated immunity using a c-Myb DNA vaccine in a mouse model of colon cancer. AB - Overexpression of the proto-oncogene c-Myb occurs in more than 80% of colorectal cancer (CRC) and is associated with aggressive disease and poor prognosis. To test c-Myb as a therapeutic target in CRC we devised a DNA fusion vaccine to generate an anti-CRC immune response. c-Myb, like many tumor antigens, is weakly immunogenic as it is a "self" antigen and subject to tolerance. To break tolerance, a DNA fusion vaccine was generated comprising wild-type c-Myb cDNA flanked by two potent Th epitopes derived from tetanus toxin. Vaccination was performed targeting a highly aggressive, weakly immunogenic, subcutaneous, syngeneic, colon adenocarcinoma cell line MC38 which highly expresses c-Myb. Prophylactic intravenous vaccination significantly suppressed tumor growth, through the induction of anti-tumor immunity for which the tetanus epitopes were essential. Vaccination generated anti-tumor immunity mediated by both CD4+ and CD8+ T cells and increased infiltration of immune effector cells at the tumor site. Importantly, no evidence of autoimmune pathology in endogenous c-Myb expressing tissues was detected as a consequence of breaking tolerance. In summary, these results establish c-Myb as a potential antigen for immune targeting in CRC and serve to provide proof of principle for the continuing development of DNA vaccines targeting c-Myb to bring this approach to the clinic. PMID- 18386001 TI - Impact of IFNalpha2b upon pSTAT3 and the MEK/ERK MAPK pathway in melanoma. AB - PURPOSE: High-dose IFNalpha2b (HDI) was established as the first effective adjuvant therapy for patients with high-risk resected melanoma more than a decade ago, but its fundamental molecular mechanism of action remains unclear. STAT3 and the mitogen activated protein kinases (MAPKs), especially ERK (extracellular signal-regulating kinase) and MEK (MAPK/ERK kinase), play roles in melanoma progression and host immunity. We have therefore evaluated STAT3 and MEK/ERK MAP kinases in patients with regional lymph node metastasis (stage IIIB) of melanoma in the context of a prospective neoadjuvant trial of HDI (UPCI 00-008). PATIENTS AND METHODS: In the context of this trial, HDI was administered daily for 20 doses following diagnostic biopsy, and prior to definitive surgery. Immunohistochemistry for pSTAT3, phospho-MEK1/2, phospho-ERK1/2, and EGFR was performed on paired fixed (nine patients) biopsies. RESULTS: HDI was found to down-regulate pSTAT3 (P = 0.008) and phospho-MEK1/2 (P = 0.008) levels significantly in tumor cells. Phospho-ERK1/2 was down-regulated by HDI in tumor cells (P = 0.015), but not in lymphoid cells. HDI down-regulated EGFR (P = 0.013), but pSTAT3 activation appeared not to be associated with EGFR expression and the MEK/ERK MAPK pathway. CONCLUSION: We conclude that HDI regulates MAPK signaling differentially in melanoma tumor cells and host lymphoid cells in vivo. STAT3 activation is independent of the EGFR/MEK/ERK signaling pathway. PMID- 18386002 TI - Minimally invasive treatment of pathological fractures of the humeral shaft. AB - Most patients with pathological fractures due to cancer metastasis have a limited life expectancy. Orthopaedic procedures, therefore, should be minimally invasive in order to avoid additional surgical morbidity. The purpose of this study was to analyse the results of minimally invasive approaches, including locked intramedullary nailing, followed by early postoperative radiation for pathological humeral shaft fractures. Twenty-four pathological fractures of the humerus diaphysis in 23 patients were treated with the prospective protocol, including antegrade unreamed intramedullary nailing and postoperative radiotherapy (20 Gy and five fractions). The patients and results of the surgery were evaluated by the Musculoskeletal Tumor Society upper extremity scoring system. All patients had a stable extremity, and the average function of 20 patients was 64% of the normal upper extremity function. Only one patient required revision surgery. The minimally invasive treatment of patients with pathological fractures of the humeral shaft with closed unreamed intramedullary nailing combined with adjuvant radiotherapy is an effective and safe procedure, even in seriously ill patients. PMID- 18386003 TI - Computer navigation in total hip replacement: a meta-analysis. AB - Proponents of navigated hip arthroplasty have suggested that it may increase the precision of acetabular component placement. We conducted a systematic review and meta-analysis to evaluate the validity of this theory. We searched, in duplicate, MEDLINE, EMBASE and the Cochrane Central Register of Controlled Trials for randomised trials comparing the use of computer navigation with the freehand technique for acetabular cup placement within the desired alignment. We assessed the methodological quality of the studies and abstracted the relevant data. Tests of heterogeneity and publication bias were performed. From the three studies included, there was no evidence of heterogeneity between studies. A total of 250 patients were entered into the analysis. The beneficial odds ratio for the number of outliers was 0.285 (95% confidence interval [CI]: 0.143 to 0.569; p < 0.001). We conclude that navigation in hip arthroplasty improves the precision of acetabular cup placement by decreasing the number of outliers from the desired alignment. PMID- 18386004 TI - Hemorrhagic cystitis in a patient receiving gemtuzumab ozogamicin for relapsed acute promyelocytic leukemia after cord blood transplantation. PMID- 18386005 TI - First detection of the codons 41-43 (-CTTTG,+A) beta-thalassemia mutation in a Chinese patient. PMID- 18386006 TI - Simvastatin-induced thrombocytopaenia: a further case and a brief on its clinical relevance. PMID- 18386007 TI - Invasive infections with a coagulase-negative staphylococcus in an immunocompromised patient: case report and review of the literature. PMID- 18386008 TI - Influence of rituximab-CHOP therapy on clinical course and autoimmune parameters in rheumatoid arthritis associated with diffuse large B cell non-Hodgkin lymphoma. PMID- 18386009 TI - Histiocytic sarcoma localised in the thyroid: a case report. PMID- 18386010 TI - Allogeneic stem cell transplantation from related and unrelated donors for aplastic anaemia in adults--a single-centre experience. AB - Aplastic anaemia (AA) is a rare bone marrow failure syndrome treated either by immunosuppressive therapy or allogeneic stem cell transplantation (SCT). At present, no randomised clinical trials evaluating both treatment options, and in particular SCT from unrelated donors, are available. We here report the clinical course and outcome of allogeneic SCT for 20 consecutive adult patients with AA. Newly diagnosed and untreated patients (n = 8) or patients pre-treated by immunosuppressive therapy (n = 12) were transplanted either from human-leukocyte antigen (HLA) identical family donors (n = 13) or matched (n = 6) and mismatched (n = 1) unrelated donors, respectively. Conditioning varied depending on donor type and included cyclophosphamide with or without anti-thymocyte globulin (ATG) and fludarabine-cyclophosphamide-ATG with or without low-dose total body irradiation. With a median follow-up of more than 40 months, all patients have had favourable outcomes with stable haematopoietic engraftment and high performance scores. Six patients developed acute (five I degrees -II degrees ; one >II degrees ) and four limited chronic graft-versus-host disease. In this group of AA patients, allogeneic SCT has proven very successful, independent of donor type and pre-treatment. Studies with greater cohorts of patients are warranted to better determine indication and timing of SCT especially from unrelated donors in AA. PMID- 18386012 TI - Detection of hypervascular malignant foci in borderline lesions of hepatocellular carcinoma: comparison of dynamic multi-detector row CT, dynamic MR imaging and superparamagnetic iron oxide-enhanced MR imaging. AB - The study object was to retrospectively compare the detection rate of hypervascular foci visualized by CT during hepatic arteriography (CTHA) in borderline nodules, which was observed upon cirrhotic livers, on dynamic MDCT, dynamic gadolinium-enhanced MR (dynamic MR), and SPIO-enhanced MR imaging. Eighty five nodules in 49 patients with cirrhosis were evaluated. When a part of the nodule showed hyperdensity relative to the surrounding areas of the nodule on CTHA, it was defined as "hypervascular focus." The relationships between the dynamic MDCT and dynamic MR and SPIO-enhanced MR imaging findings of these foci were analyzed using X(2) test. Hypervascular foci were detected in 17 (53%) of 32 on the arterial dominant phase of dynamic MDCT, in 19 (37%) of 51 on the arterial dominant phase of dynamic MR and in 6 (26%) of 23 on SPIO-enhanced MR imaging. Arterial dominant phase of dynamic MDCT demonstrated a significantly higher detection rate of hypervascular foci less than 5 mm in diameter than did dynamic and SPIO MR imaging (p<0.05). Hypervascular foci in borderline nodules could be better visualized by dynamic MDCT than by gadolinium- and SPIO-enhanced MR imaging. Dynamic MDCT is recommended for the follow-up examination of hypovascular borderline lesions. PMID- 18386011 TI - From single cells to tissue architecture-a bottom-up approach to modelling the spatio-temporal organisation of complex multi-cellular systems. AB - Collective phenomena in multi-cellular assemblies can be approached on different levels of complexity. Here, we discuss a number of mathematical models which consider the dynamics of each individual cell, so-called agent-based or individual-based models (IBMs). As a special feature, these models allow to account for intracellular decision processes which are triggered by biomechanical cell-cell or cell-matrix interactions. We discuss their impact on the growth and homeostasis of multi-cellular systems as simulated by lattice-free models. Our results demonstrate that cell polarisation subsequent to cell-cell contact formation can be a source of stability in epithelial monolayers. Stroma contact dependent regulation of tumour cell proliferation and migration is shown to result in invasion dynamics in accordance with the migrating cancer stem cell hypothesis. However, we demonstrate that different regulation mechanisms can equally well comply with present experimental results. Thus, we suggest a panel of experimental studies for the in-depth validation of the model assumptions. PMID- 18386013 TI - Radiological profile of anemia on unenhanced MDCT of the thorax. AB - Our aim was to investigate the diagnostic value of unenhanced MDCT in anemic patients. Fifty consecutive patients with proven anemia and 50 nonanemic matched group for age, sex and body mass index were evaluated. In either group, hemoglobin levels were assessed no more than 24 h from an unenhanced CT of the thorax. For each patient, the presence of a hyperattenuating aortic wall (aortic ring sign) and/or dense interventricular septum (subjective parameters) were identified by two radiologists who were blinded to the laboratory findings. Furthermore, the aortic CT attenuation values (objective parameter) were also obtained and correlated with the hemoglobin levels. The sensitivity and specificity in detecting anemia were calculated for each variable, and ROC analysis was generated for subjective and objective parameters. Subjective image analysis revealed that the aortic ring sign was more sensitive than the interventricular septum sign for anemia detection (84% vs. 72%), whereas this latter sign was more specific (100% vs. 92%). A good correlation (r = 0.60) was observed between the aortic CT attenuation values and the hemoglobin levels in the whole study population. Using a threshold of < or = 35 HU for anemia diagnosis, the sensitivity and specificity of aortic CT attenuation value were 84% and 94%, respectively, with the largest area under the curve (0.89) among all diagnostic criteria. However, the best trade-off between sensitivity (80%) and specificity (100%) was obtained from combining both subjective and objective analysis. Interpreting anemia upon unnenhanced MDCT of the thorax is quite feasible. A diagnostic approach that considers both subjective and objective analysis offers the best trade-off between sensitivity and specificity. PMID- 18386014 TI - Quadriceps tendinosis and patellar tendinosis in professional beach volleyball players: sonographic findings in correlation with clinical symptoms. AB - The purpose was to assess quadriceps and patellar tendinosis in professional beach volleyball players and to correlate ultrasound findings with clinical symptoms. During a grand-slam beach volleyball tournaments all 202 athletes (100 men and 102 women) were invited to participate at this study. Sixty-one athletes (38 male, mean age 29.6, 23 female, mean age 27.1) were included. The dominant leg was right in 51 (84%) and left in ten athletes (16%). Lysholm knee score and pain during the game was assessed using a visual analogue scale. Sonography of the quadriceps tendon and the patellar tendon was performed by a blinded sonographer. Sonographic findings were compared between both legs and correlated to clinical findings using a regression analysis. Quadriceps tendinosis was diagnosed in 13 (21%, dominant leg)/21 (34%, non-dominant leg), patellar tendinosis in 13(21%)/18(30%). Only sonographic findings at the quadriceps tendon were significantly associated with pain: thickness of the quadriceps tendon (mean diameter 6.9 mm/7.1 mm, significant for both legs P = 0.011/P = 0.030), abnormal echo texture (11/16; P = 0.001/P = 0.228), areas with positive power Doppler signals (mean number 0.3/0.4; P = 0.049/0.346), calcifications (mean number: 0.9/1.1; P = 0.021/0.864). A relationship between findings at patellar tendon was not found. Quadriceps tendinosis is as common as patellar tendinosis in professional beach volleyball players. Thickening and structure alteration of the quadriceps tendon is associated with anterior knee pain during beach volleyball. PMID- 18386015 TI - Optional inferior vena caval filters: where are we now? AB - With the advent of newer optional/retrievable inferior vena caval filters, there has been a rise in the number of filters inserted globally. This review article examines the currently available approved optional filter models, outlines the clinical indications for filter insertion and examines the expanding indications. Additionally, the available evidence behind the use of optional filters is reviewed, the issue of anticoagulation is discussed and possible future filter developments are considered. PMID- 18386016 TI - Polarization vision in crayfish motion detectors. AB - Motion detector interneurons were examined to determine their responsiveness to the motion of polarized light images (i.e. images segmented by spatial variations in e-vector angle). Computer generated images were displayed as intensity contrasts or polarization contrasts on a modified LCD projection panel. The stimuli included the motion of a single stripe (45 degrees -55 degrees /s) and the global motion of a square wave grating (3.3 degrees /s). Neurons were impaled in the medulla interna. Of the neurons which exhibited a directional response to the motion of intensity contrast stimuli, about 2/3 were also directional in the response to polarized light images. Transient (nondirectional) stimuli included looming and jittery motions. The responses to the transient motions of the polarized light images were roughly comparable to those elicited by intensity contrast. The results imply that behavioral responses to polarized light images (i.e. optokinetic and defense reflexes) may have a basis in the polarization sensitivity and synaptic organization of the medulla interna. PMID- 18386017 TI - Song motor control organizes acoustic patterns on two levels in Bengalese finches (Lonchura striata var. domestica). AB - Based on statistical analyses of song sequences, Bengalese finch (Lonchura striata var. domestica) songs do not show unvarying motif repetition as has been found in zebra finches (Taeniopygia guttata). Instead, there are variations of partially stereotyped sequences of song syllables. Although these stereotyped sequences consist of multiple syllables, in most cases these syllables occur together. To examine whether such structures really exist as a vocal production unit, we subjected singing birds to a light flash and determined when the stimulus stopped the songs. When light interruptions were presented within the statistically stereotyped sequences, the subsequent syllables tended to be produced, whereas interruptions presented during the statistically variable sequences tended to cause instantaneous song termination. This suggests that the associations among the song syllables that compose the statistically stereotyped sequences are more order dependent than those for the statistically variable sequences, and the tolerances of syllable pairs to visual interruptions are consistent with the statistical song structures. Additionally, following interruptions, several types of song sequence variations were observed that had not been previously reported. These phenomena might be caused by various effects of the visual stimulus on the hierarchical motor control program. PMID- 18386018 TI - Mechanics of the exceptional anuran ear. AB - The anuran ear is frequently used for studying fundamental properties of vertebrate auditory systems. This is due to its unique anatomical features, most prominently the lack of a basilar membrane and the presence of two dedicated acoustic end organs, the basilar papilla and the amphibian papilla. Our current anatomical and functional knowledge implies that three distinct regions can be identified within these two organs. The basilar papilla functions as a single auditory filter. The low-frequency portion of the amphibian papilla is an electrically tuned, tonotopically organized auditory end organ. The high frequency portion of the amphibian papilla is mechanically tuned and tonotopically organized, and it emits spontaneous otoacoustic emissions. This high-frequency portion of the amphibian papilla shows a remarkable, functional resemblance to the mammalian cochlea. PMID- 18386019 TI - Comparison of musculoskeletal and urological functional outcomes in patients with bladder exstrophy undergoing repair with and without osteotomy. AB - It is controversial as to whether osteotomy, by restoring a more normal pelvic anatomy, might improve the final outcome of bladder exstrophy (BE) repairs. We compared the functional orthopaedic and urological outcomes in BE patients treated with and without osteotomy. Orthopaedic and urological outcomes were compared in eight BE patients treated with osteotomy and six BE patients treated without osteotomy. Orthopaedic evaluation included an assessment of pubic bones dissymmetry, bending of the spine, presence of Trendelenburg or Thomas sign, and presence of out-toeing. Pubic diastasis was ruled out on a plain X-ray of the pelvis. A Pediatric Orthopedic Society of North America (POSNA) questionnaire was administered to every child or his/her caregiver to assess functional outcome. Urological evaluation included an assessment of required continence surgeries and of contemporary continence status. All patients presented a pubic diastasis. This was in median 49 (24-66) mm in patients treated without osteotomy and 42 (25-101) mm in those treated with osteotomy (p = 0.3). There was no difference either in the orthopaedic outcome or in any features of the POSNA questionnaire between groups. Neither was there a difference in the final continence rate nor in the number of additional continence procedures required. Although osteotomy is an essential step in the treatment of many BE patients in order to achieve a tension free closure of the abdominal wall and bladder, our preliminary results suggest that it does not improve the eventual orthopaedic or urological outcomes of BE. PMID- 18386020 TI - The spectrum of anorectal malformations in Africa. AB - Anorectal malformations (ARM) remain a significant birth defect with geographic variation in incidence, individual phenotypes and regional geographic subtypes. Although early studies indicated a low incidence in Black patients, there is a great paucity of knowledge as to the types, frequency and incidence of ARMs encountered in the African continent and their associated anomalies. Current evidence suggests a significant clinical load. This study set out to evaluate ARM in Southern and other parts of Africa to define the clinical load of ARM. We retrospectively collected data on 1,401 ARM patients from six South African Paediatric Surgical units plus representative samples from five other African countries from West, Central and Southern Africa. Data included ethnic group, age, gender as well as the anatomical pathology, classification and presence or absence of associated anomalies. ARM lesions classified by the Wingspread classification plus an analysis of fistula position was carried out in evaluable cases for purposes of comparison. South African centres reported a higher prevalence of cloacae and vestibular fistulae, whereas rectovaginal, recto prostatic and anorectal malformation without fistula were more prevalent in the Northern African group. 76% of 1,401 patients were ethnically Black African [gender ratio = 2 (vs gender ratio 1.38 overall)] and 49.8% were "low" lesions (Wingspread classification). High or intermediate lesions were mostly males (72%). Anal stenosis was most prevalent in black males and non-Black females. Fistulae were identified in 95% with 682 (52%) being low (perineal/covered anus/vestibular) fistulae. Perineal fistulae had a male predilection (n = 260; 20%), whereas vestibular fistulas (n = 416; 32%) was strikingly frequent in black females (55%). Of the remainder, 15 fistulae were rectovesical (1.2%), 544 recto urethral or prostatic (42%), 16 recto-vaginal (1.2%). In addition, there were 43 cloacal lesions (3.3%). Isolated rare ARM anomalies included "Pouch" colon (2) and H-type fistula (2). Isolated lesions occurred in 81% but 163 associated anomalies were identified in 114 patients. These included chromosomal lesions (10), genito-urinary anomalies (50), genital (16), cardiac (31), skeletal anomalies (33), gastro-intestinal malformations (28). Other anomalies included CNS anomalies (14), anterior abdominal wall defects (2) and facial (8) abnormalities and neuroblastoma (1). The ARM is not uncommon in Black African populations and constitutes a significant clinical load to surgical practice in Africa. Ethnic differences appear to exist and vestibular fistulae predominate in black females. Cloaca (3.3%) did appear to be more prevalent. Isolated lesions are frequent but the types of associated anomalies appear similar to other series except chromosomal syndromes. This study illustrates the need for more objective data from developing countries to assess geographical differences. PMID- 18386022 TI - Operative management for atlantoaxial instability in case of bilateral high riding vertebral artery. AB - INTRODUCTION: In case of atlantoaxial instability dorsal C1-C2 screw fixation is the most appropriate procedure to get immediate and sufficient three-point fixation. However, this method cannot be performed in bilateral high-riding vertebral arteries because of high risk of VA injury. CASE DESCRIPTION: In a 66 year-old woman who presented with neck pain due to atlantoaxial instability bilateral high-riding VAs were detected in CT. To avoid VA injury a dorsal stabilization procedure with a hook-rod system and interspinous autologous iliac crest graft was performed. The intra- and postoperative courses had no complications and the hook-rod construction clinically led to immediate postoperative stability. After 24 months the patient was painfree. CONCLUSIONS: In case of atlantoaxial instability and bilateral high-riding VAs dorsal C1-C2 fixation procedure with a hook-rod system and interspinous bone grafting seems to be an appropriate alternative surgical technique and can get immediate and clinically sufficient stability with no risk of VA injury. PMID- 18386021 TI - Peroxiredoxin 6 in human brain: molecular forms, cellular distribution and association with Alzheimer's disease pathology. AB - Peroxiredoxin 6 is an antioxidant enzyme and is the 1-cys member of the peroxiredoxin family. Using two-dimensional electrophoresis and Western blotting, we have shown for the first time that, in human control and brain tissue of patient's with Alzheimer's disease (AD), this enzyme exists as three major and five minor forms with pIs from 5.3 to 6.1. Using specific cellular markers, we have shown that peroxiredoxin 6 is present in astrocytes with very low levels in neurons, but not detectable in microglia or oligodendrocytes. In control brains, there was a very low level of peroxiredoxin 6 staining in astrocytes that was confined to a "halo" around the nucleus. In AD, there were marked increases in the number and staining intensity of peroxiredoxin 6 positive astrocytes in both gray and white matter in the midfrontal cortex, cingulate, hippocampus and amygdala. Confocal microscopy using antibodies to A beta peptide, tau and peroxiredoxin 6 showed that peroxiredoxin 6 positive astrocytes are closely involved with diffuse plaques and to a lesser extent with neuritic plaques, suggesting that plaques are producing reactive oxygen species. There appeared to be little astrocytic response to tau containing neurons. Although peroxiredoxin 6 positive astrocytes were seen to make multiple contacts with tau positive neurons, there was no intraneuronal colocalization. In brain tissue of patients with AD, many blood vessels exhibited peroxiredoxin 6 staining that appeared to be due to the astrocytic foot processes. These results suggest that oxidative stress conditions exist in AD and that peroxiredoxin 6 is an important antioxidant enzyme in human brain defenses. PMID- 18386023 TI - Effects of physical and chemical treatments upon biophysical properties and micro relief of human skin. AB - The aim of the present study was to determine the attendant effects of physical (tape-stripping) and chemical (three commercial hydrating formulations) treatments upon biophysical and micro-relief properties of human skin. In the first set of experiment, the effects of tape-stripping onto human stratum corneum (SC) biophysical and micro-relief properties were assessed in nine volunteers. Transepidermal water loss (TEWL), skin hydration and micro-relief parameters (including total length of the lines in mm per mm(2); total surface in %; roughness of the skin measured in gray level (Ra); maximum profile valley (Rv) depth; maximum profile peak height (Rp); maximum height (Rt), peak density (Pc) and coefficient of anisotropy) were determined by using SkinEvidence Pro after subsequent tape-stripping of SC. The relevance of roughness determination as gray level by SkinEvidence Pro was confirmed by using surface roughness standards. In the second set of experiment, the effectiveness of three commercial hydrating formulations onto human SC biophysical parameters and micro-relief properties were assessed in six volunteers. TEWL, hydration and micro-relief parameters were assessed onto pre-treated acetone skin and then treated by three commercial hydrating formulations after 2, 4 and 6 h skin exposure. A linear relation between hydration and cutaneous parameters (total length of the lines, Ra and Rp) as function of SC removed was shown. Skin barrier properties evaluated by TEWL measurements, were not modified by topical formulations. However, skin treated by topical formulations showed slightly higher hydration than the one determined in control group, while micro-relief parameters were not modified. In this study was showed that biophysical and micro-relief parameters were closely related in tape stripping experiment. Efficiency of topical formulations was suggested upon skin hydration but not onto skin micro-relief and barrier function recovering. From both experiments, it appears that different mechanisms relating to skin hydration and potential modification of cutaneous micro-relief were suggested. PMID- 18386024 TI - The expression of syndecan-1 in psoriatic epidermis. AB - Psoriasis is a chronic inflammatory skin disease characterized by exaggerated keratinocyte proliferation. Current opinion indicates that psoriasis is driven by T cell-mediated immune responses targeting keratinocytes. However, psoriasis cannot be explained solely on the basis of T-cell activation, and it is likely that an intrinsic alteration in epidermal keratinocytes plays a very important role in disease expression. Syndecans comprise a major family of cell surface heparan sulfate proteoglycans. Several studies indicate their role in adhesion, cell-extracellular matrix interactions, migration, keratinocyte proliferation and differentiation, inflammation, and wound healing. To determine the expression of syndecan-1 in psoriasis, skin samples from 29 patients with fully developed psoriasis and skin samples from 14 healthy volunteer persons with no personal or family history of psoriasis were immunohistochemically examined using monoclonal antibody against syndecan-1. The expression of syndecan-1 was analyzed in whole mount section of psoriatic and non-psoriatic skin biopsies under high magnification (400x). In addition, the intensity and topography of reaction in the cell, as well as localization of positive cells in the epidermis were evaluated. Strong syndecan-1 reactivity in epidermal cells in all non-psoriatic and psoriatic samples was observed. Statistical analysis showed no significant differences between two analyzed groups (P > 0.05). In normal skin syndecan-1 was expressed in full thickness of the epidermis. The strongest reaction was observed in membranes and intercellular junctions of spinous and granular layer while basal cells showed weaker expression that was confined to cytoplasm. In psoriatic skin syndecan-1 was expressed in the membrane and intercellular junction of cells located in thickened and elongated rete ridges of the epidermis. The strongest reaction was in basal and suprabasal layers and expression diminished through spinous layer. Cells in spinous layer lose syndecan-1 expression, which is opposite pattern to normal skin. Our results suggest that aberrant skin expression of syndecan-1 may be involved in the development of psoriasis. PMID- 18386025 TI - Effects of acetylcholine chloride on intracellular calcium concentration of cultured sweat gland epithelial cells. AB - In order to isolate and culture the sweat gland epithelial cells in vitro and to study the effects of acetylcholine (ACh) on intracellular calcium concentration ([Ca(2+)](i)) of cultured sweat gland epithelial cells, the following methods were used. First, repeated shearing and neutral red staining made the sweat glands pop out from subcutaneous tissues. Then, transferpettor was used to pick up the glands, which were cultured in Epilife after type II collagenase digestion. The molecular characterization of primary cultured sweat gland epithelial cells was shown by immunocytochemistry. The [Ca(2+)](i) was examined by confocal laser scanning microscopy (CLSM) with the Ca(2+)-sensitive dye Fura 3 AM, when ACh was added to the primary cells and the first passage cells. In the results, the established method yielded comparatively more sweat glands, and the primary and first passage epithelial cells developed well in Epilife. The primary epithelial cells were positive to anti-EMA, anti-CK and anti-CK7. After the ACh was added, when the medium with high calcium (2 mmol/L) was applied, the calcium channel of both primary and first passage cells opened and significant [Ca(2+)](I )increase was observed; when the medium with no calcium was applied, no significant [Ca(2+)](i )increase was observed. So, it is a good method to isolate sweat glands by repeated shearing and transferpettor picking, and the culture mediums of keratinocytes, like Epilife, can be used to culture the sweat glands epithelial cells. In both the cultured primary and first passage cells, when stimulated by ACh, the calcium channel opened, which induced an increase in [Ca(2+)](i), similar to the cells in vivo. PMID- 18386026 TI - Surfactant proteins in inflammatory skin diseases: controlled study. AB - Surfactant proteins (SP) have recently been reported to be expressed in human skin tissue. SP is thought to play an essential role in the firstline defense of skin. In this study, we aimed to investigate if the SP may play a role in inflammatory skin diseases. Seven volunteers with psoriasis (n = 3), atopic dermatitis (n = 2), lichen planus (n = 1) and Behcet's disease (n = 1) participated in the study. Biopsies from each lesion and adjacent (approximately 2 cm distant) normal-appearing skin in patients were performed. Expression and localization of the SP-A, -B, -C, and -D in fresh tissues were studied by an immunohistochemical technique. In all patients, there was a weak cytoplasmic staining with SP-A and SP-D and nuclear staining with SP-B and SP-C in the epidermis of normal-appearing skin samples. However, epidermal staining with SP was observed to be stronger in all lesional samples. In addition, there was a prominent staining in inflammatory cells infiltrating dermis. This expression represents a previously unknown immunologic response in the inflammatory skin diseases and may represent an important step in the pathogenesis of these disorders. PMID- 18386027 TI - SERPINE1 (PAI-1) is deposited into keratinocyte migration "trails" and required for optimal monolayer wound repair. AB - Cutaneous tissue injury, both in vivo and in vitro, initiates activation of a "wound repair" transcriptional program. One such highly induced gene encodes plasminogen activator inhibitor type-1 (PAI-1, SERPINE1). PAI-1-GFP, expressed as a fusion protein under inducible control of +800 bp of the wound-activated PAI-1 promoter, prominently "marked" keratinocyte migration trails during the real-time of monolayer scrape-injury repair. Addition of active recombinant PAI-1 to wounded wild-type keratinocyte monolayers as well as to PAI-1(-/-) MEFs and PAI 1(-/-) keratinocytes significantly stimulated directional motility above basal levels in all cell types. PAI-1 expression knockdown or antibody-mediated functional inhibition, in contrast, effectively attenuated injury repair. The defect in wound-associated migratory activity as a consequence of antisense mediated PAI-1 down-regulation was effectively reversed by addition of recombinant PAI-1 immediately after scrape injury. One possible mechanism underlying the PAI-1-dependent motile response may involve fine control of the keratinocyte substrate detachment/re-attachment process. Exogenous PAI-1 significantly enhanced keratinocyte spread cell "footprint" area while PAI-1 neutralizing antibodies, but not control non-immune IgG, effectively inhibited spreading with apoptotic hallmarks evident within 24 h. Importantly, PAI-1 not only stimulated keratinocyte adhesion and wound-initiated planar migration but also rescued keratinocytes from plasminogen-induced substrate detachment/anoikis. The early transcriptional response of the PAI-1 gene to monolayer trauma and its prominence in the injury repair genetic signature are consistent with its function as both a survival factor and regulator of the time course of epithelial migration as part of the cutaneous injury response program. PMID- 18386028 TI - Reliability and validity of a bioimpedance measurement device in the assessment of UVR damage to the skin. AB - Ultraviolet radiation (UVR) is the primary cause of skin cancers. However, it is difficult to evaluate the amount of UVR absorbed into the skin retrospectively. Therefore, objective and non-invasive quantitative method would be valuable for epidemiological UVR exposure assessment. Photodamage reduces the amount of bound water in the skin, and thus, measuring the skin's dielectric constant can provide an opportunity for assessing the cumulative UVR exposure. The purpose of the study was to assess the reliability and validity of the bioimpedance device, Moisture Meter-D. The measurements were performed on 100 subjects at three separate measurement times. A questionnaire was used to obtain information on the host factors and on the past UVR exposure. The biological samples, to determine the elastin proportion of the dermis, were collected. Some long-term as well as seasonal variations in the dielectric constants were detected. Also, a weak relationship between the dielectric constant and the UVR exposure indicators and host factors was observed. The MoistureMeter-D appears not to measure structural alterations in the skin caused by photodamage, and thus it is not a valid instrument for the assessment of photodamage, i.e., past UVR exposure. PMID- 18386029 TI - The relationship between urinary tract infection during pregnancy and preeclampsia: causal, confounded or spurious? AB - BACKGROUND: Preeclampsia is a major cause of maternal morbidity, although its precise etiology remains elusive. A number of studies suggest that urinary tract infection (UTI) during the course of gestation is associated with elevated risk for preeclampsia, while others have failed to prove such an association. In our medical center, pregnant women who were exposed to at least one UTI episode during pregnancy were 1.3 times more likely to have mild preeclampsia and 1.8 times more likely to have severe preeclampsia as compared to unexposed women. Our results are based on univariate analyses and are not adjusted for potential confounders. OBJECTIVE: This editorial aims to discuss the relationship between urinary tract infection and preeclampsia, as well as examine the current problems regarding the interpretation of this association. CONCLUSION: Although the relationship between UTI and preeclampsia has been demonstrated in studies with various designs, carried-out in a variety of settings, the nature of this association is unclear. By taking into account timeline, dose-response effects, treatment influences, and potential confounders, as well as by neutralizing potential biases, future studies may be able to clarify the relationship between UTI and preeclampsia by determining if it is causal, confounded, or spurious. PMID- 18386030 TI - Removal of enormous bilateral mucinous cystadenomas of the ovaries with abdominal plastic reconstruction. AB - INTRODUCTION: Bilateral mucinous cystadenoma of the ovary are extremely rare. These tumors are benign and might lead to abdominal distension, if no secondary symptoms occur and patient delay the consultation of physicians. CASE: A 60-year old patient was admitted to the internal medicine department for constipation and dyspnoea. Transabdominal punctions were performed because of suspected ascites. Finally the patient was transferred to the gynecological department after forced diuresis did not reduce the suspected ascites and a CT scan showed large intraabdominal masses. Tumor excision was initiated and final histology revealed bilateral mucinous cystadenoma of the ovaries. DISCUSSION: Huge intraabdominal tumors that almost double a patient's body weight can hardly be malign. Total tumor excision is necessary because the heterogeneous composition requires careful examination by pathologists to rule out borderline tumors and non invasive carcinomas. After tumor excision an abdominal wall reconstruction might be necessary because of the laxity and redundancy of the skin. PMID- 18386032 TI - Radon survey in dwellings of Gansu, China: the influence of thoron and an attempt for correction. AB - Alpha track detectors used in a previous investigation of the US National Cancer Institute and the China Ministry of Health on indoor radon ((222)Rn) in Gansu, China, proved to be influenced by (220)Rn (thoron), thus overestimating the (222)Rn level. Therefore, the detector was improved used in the previous survey. The new detectors allow discrimination between the two isotopes without any disturbance of the (222)Rn measurement. With this detector, a semi-annual study was conducted in 49 traditional dwellings of a village in Gansu. The arithmetic (AM) and geometric (GM) mean (222)Rn concentrations were 120 +/- 61 and 105 Bq m( 3) (with geometric standard deviation GSD = 1.8), respectively, while the mean (220)Rn concentrations at 2.5 cm wall distance were 430 +/- 210 Bq m(-3) (AM) and 350 Bq m(-3) (GM) with GSD = 2.3. The high thoron concentrations demonstrate the importance of the (220)Rn contribution to radiation exposure, in the investigated area. The actual level of indoor (222)Rn was about three times lower than that in the previous investigation which was affected by (220)Rn. A correction method for the radon results of the previous study is proposed, which provides (222)Rn and (220)Rn values comparable with those obtained in the study presented here. PMID- 18386033 TI - An epidemic of fatal 3-methylfentanyl poisoning in Estonia. AB - An exceptional epidemic of poisonings due to the highly potent opioid designer drug 3-methylfentanyl (TMF) was revealed among Estonian drug users in 2005-2006 by post-mortem forensic toxicology. Quantitative analysis of cis-TMF, trans-TMF, and fentanyl was performed by liquid chromatography-tandem mass spectrometry. Comprehensive toxicological analysis was performed using a multi-technique approach. The number of TMF-related fatal accidental poisonings identified was 46 and 71 for 2005 and 2006, respectively. The proportion of male victims was 91.5% and the mean age of all victims was 26 years at death. TMF was used predominantly by intravenous injection. There was no significant difference in the blood concentrations of cis-TMF and trans-TMF between pure TMF poisonings and mixed TMF poisonings. The mean combined concentration of TMF stereoisomers among pure TMF cases (1.9 microg/l) was more than ten times lower than the mean fentanyl concentration in fentanyl-related fatalities. Concomitant use of other drugs involved alcohol, amphetamines, benzodiazepines, and cannabis, but very rarely other opioids. PMID- 18386034 TI - Episcleral melanoma as a complication of ocular melanocytosis. AB - BACKGROUND: Primary episcleral melanoma is exceedingly rare. We report a case of primary episcleral melanoma in an eye with ocular melanocytosis. METHODS: Interventional case report with presentation of clinical photographs and histopathology. RESULTS: A 62-year-old man with long-standing ocular melanocytosis presented with a new, raised, darkly pigmented lesion on the episclera. Clinical examination and ultrasound biomicroscopy showed no evidence of intraocular mass or nevus. Lamellar scleroconjunctivectomy of the lesion was performed, and histopathologic analysis revealed episcleral melanoma with no evidence of intra- or extra-ocular extension. He was later treated with cryotherapy to the surgical bed. Continued observation shows no evidence for intraocular tumor or recurrence of episcleral tumor. CONCLUSIONS: Primary episcleral melanoma is a previously unreported complication of ocular melanocytosis, and should be considered in the differential diagnosis in patients with ocular melanocytosis and pigmented nodules on the ocular surface. PMID- 18386035 TI - Structure-function relationship in ocular hypertension and glaucoma: interindividual and interocular analysis by OCT and pattern ERG. AB - BACKGROUND: Pattern electroretinogram (PERG) and optical coherence tomography (OCT) represent objective probes to investigate respectively the function of retinal ganglion cells and their structure as retinal nerve fiber layer (RNFL) thickness. We examined interindividual (II) correlations of PERG amplitude and RNFL thickness, as well as correlations between interocular (IO) differences in both measures, in ocular hypertension (OHT) and early glaucoma (EG) patients. METHODS: Thirty-one OHT, 34 EG (mean deviation: -1 to -6 dB) and 16 age-matched controls were examined in both eyes. Participants had clear optical media, no or moderate refractive errors and no concomitant ocular or systemic diseases. PERGs were elicited by counterphased (16.28 reversals/second) gratings (1.6 cycles/degree spatial frequency). The Fourier isolated 2nd harmonic PERG amplitude and phase were measured. RNFL thickness was quantified by means of OCT Stratus according to a standard protocol. Average, superior and inferior RNFL thicknesses were considered. RESULTS: Mean PERG amplitude was decreased (p < 0.01) in both OHT and EG patients compared to controls. Mean RNFL thicknesses were reduced (p < 0.01) in EG patients compared to both OHT and controls. In OHT patients, PERG amplitude did not correlate significantly with RNFL thickness in both II and IO analysis. In EG patients, PERG amplitude was positively correlated with RNFL thickness in both II (p < 0.005) and IO (p < 0.001) analysis. The slope of the correlation predicted that PERG losses exceeded systematically RNFL losses when the latter were between 0 and -0.25 log units. CONCLUSIONS: Both II and IO analyses revealed a lack of structure-function relationship in OHT, suggesting that, at this disease stage, PERG losses appear to affect primarily retinal/optic nerve head function. In EG they reflect both dysfunction and RNFL loss. PMID- 18386036 TI - Prolonged retinal arteriovenous passage time is correlated to ocular perfusion pressure in normal tension glaucoma. AB - BACKGROUND: The pathogenesis of normal tension glaucoma (NTG) might be related to impaired autoregulation of ocular blood flow. The purpose of the study is to evaluate retinal haemodynamics by fluorescein angiography and to correlate arteriovenous passage times (AVP) with ocular perfusion pressure in patients with NTG and controls. METHODS: Thirty-five patients with NTG without any topical treatment (mean age 53 +/- 11 years) and 35 age-matched controls (mean age 53 +/- 11 years) were included in this study. Retinal AVP was assessed by video fluorescein angiography using a scanning laser ophthalmoscope (Rodenstock, Germany). Dye dilution curves of temporal superior and inferior arterioles and venules were evaluated by digital image analysis. AVP was correlated to mean arterial blood pressure and ocular perfusion pressure. RESULTS: The AVP was significantly prolonged in patients with NTG compared to controls (1.82 +/- 0.57 versus 1.42 +/- 0.46, p = 0.002). Patients with NTG and controls showed no significant differences in intraocular pressure, mean arterial pressure and mean and diastolic ocular perfusion pressure. The AVP was significantly correlated to mean arterial pressure and mean and diastolic ocular perfusion pressure in patients with NTG (r = -0.54; p = 0.0006, r = -0.51; p = 0.002, r = -0.49, p = 0.002), but not in controls (r = -0.21; p = 0.23, r = -0.19; p = 0.27, r = 0.02, p = 0.93). CONCLUSIONS: Patients with NTG exhibit prolonged retinal AVP compared to controls. A significant correlation of retinal haemodynamics to mean arterial blood pressure and ocular perfusion pressure might reflect impaired autoregulation in NTG. PMID- 18386037 TI - Choroidal plasmacytoma in a patient with multiple myeloma. Diagnosis by fine needle aspiration biopsy. AB - BACKGROUND: To report a case of a secondary plasmacytoma of the choroid. METHODS: Interventional case report with cytopathologic correlation. RESULTS: A 61-year old male with a breast nodule and a 2-year history of multiple myeloma developed blurred vision and was found to have three confluent, ill-defined areas of choroidal thickening in the left eye. Cytopathology and immunohistochemistry of a transocular fine-needle aspiration biopsy of the largest lesion revealed atypical plasma cells diagnostic of plasmacytoma. Following external beam radiotherapy the choroidal tumors resolved completely. CONCLUSION: Secondary plasmacytoma, despite its rarity, should be included in the differential diagnosis of amelanotic choroidal tumors, particularly in patients with multiple myeloma. Radiotherapy is an effective treatment. PMID- 18386038 TI - Immunohistochemical evaluation of the extracellular matrix in trabecular meshwork in steroid-induced glaucoma. AB - BACKGROUND: To immunohistochemically examine the localization of the extracellular matrix (ECM) in the trabecular meshwork (TM) in steroid-induced glaucoma (SG) specimens. METHODS: We morphologically and immunohistochemically examined six trabecular tissues from three eyes with SG, two with primary open angle glaucoma (POAG), and one without glaucoma. For the morphological study, the ultra-microtome sections were observed using an electron microscope. For the light microscopic immunohistochemical analyses, frozen sections were stained by the avidin-biotin complex method using anti-type IV collagen, anti-type VI collagen, anti-heparan sulfate proteoglycan (HSPG), anti-fibronectin or anti myocilin (MYOC) antibody. RESULTS: The morphological examinations revealed accumulations of basement membrane-like and fine fibrillar-like materials in the outer TM of SG specimen. Type IV collagen, HSPG and fibronectin antibodies in SG specimens showed a greater degree of staining in the outer TM in comparison to the POAG and non-glaucomatous specimens; in contrast, the other antibodies including the type VI collagen and MYOC, did not. CONCLUSIONS: The localization of ECMs in the TM is different in SG in comparison to that in POAG patients. PMID- 18386039 TI - Bevacizumab in the treatment of idiopathic macular telangiectasia. AB - BACKGROUND: To describe functional and morphological long-term follow-up results in patients with idiopathic macular telangiectasia (IMT) treated with intravitreal bevacizumab. METHODS: Retrospective case series of three consecutive male patients with IMT who were treated with intravitreal bevacizumab injections. Best corrected visual acuity (BCVA) as well as fluorescein angiography (FA) and optical coherence tomography (OCT) were monitored over the period of up to 12 months. RESULTS: Single intravitreal bevacizumab injection resulted in a marked increase in BCVA from 20/50 to 20/20 in the patient with type 1 (aneurysmal) IMT during the first 4 weeks. Late-phase leakage on FA and cystoid macular oedema on OCT decreased significantly. This was sustained over the whole follow-up period of 12 months. In contrast, in the two patients with type 2 (perifoveal) IMT, leakage on FA decreased likewise, but this was not accompanied by an increase in BCVA despite triple injections. Small cystic changes seen on OCT remained unchanged. CONCLUSION: Patients with type 1 IMT with pronounced macular oedema on OCT may benefit from intravitreal bevacizumab injections, showing functional as well as morphological improvement, while patients with type 2 IMT with minimal cystic changes on OCT do not show functional improvement despite repeated injections. PMID- 18386040 TI - Correlation between foveal structure and visual outcome following macular hole surgery: a spectral optical coherence tomography study. AB - PURPOSE: The aim of this paper is to evaluate the retinal structure after macular hole surgery and to study the correlation of structural findings with final functional outcomes, using high-speed, high-resolution spectral optical coherence tomography (SOCT). METHODS: Sixty-eight eyes of 60 patients with full-thickness macular holes were included in the study. All patients underwent pars plana vitrectomy with trypan blue staining and internal limiting membrane (ILM) peeling. Patients were evaluated by SOCT, with 6 mum axial and 12-18 mum transverse resolution and three-dimensional images of the retina. RESULTS: There were four different types of macular hole closure: U-shape, V-shape, irregular and flat/open. The following retinal abnormalities were observed in postoperative scans: photoreceptor irregularities, lack of photoreceptors (photoreceptor defect), cysts in outer retinal layers, nerve fiber layer defects, lesion of all retinal layers, and RPE defects. It was possible to evaluate photoreceptor defects on a three-dimensional image. Retinal thickness in the fovea was also measured. CONCLUSIONS: Because of excellent resolution, SOCT is capable of visualization of retinal defects after macular hole surgery. Three-dimensional examination is adequate for evaluation of photoreceptor defects. Good postoperative visual acuity is correlated with U-shape closure, normal foveal thickness and absence of photoreceptor layer defects. PMID- 18386041 TI - Anterior ischemic optic neuropathy due to giant cell arteritis with normal inflammatory markers. AB - BACKGROUND: In anterior ischemic optic neuropathy (AION), it is important not to miss the diagnosis of giant cell arteritis (GCA) because this requires immediate steroid treatment to prevent involvement of the second eye and possible blindness. A missed diagnosis also might lead to fatal systemic complications. MATERIALS AND METHODS: Observational case report. RESULTS: A 79-year-old woman noticed decreased visual and visual field loss in the right eye. At presentation, right visual acuity was 10/20 (ETDRS chart 2000). There was a right relative afferent pupillary defect of 0.6 log units. Asked for symptoms of GCA she complained about temporal and occipital headache, jaw claudication combined with malaise, and myalgia of the upper limbs. Laboratory tests showed normal inflammatory markers. Repeated tests confirmed ESR and CRP to be within the normal range. GCA being suspected, ultrasound of the superficial temporal arteries and temporal artery biopsy were performed unilaterally on the right side. Histology showed a chronic inflammatory cell infiltrate consistent with active GCA. The patient was treated with high-dose corticosteroids (250 mg methylprednisolone, three times/day, initially) and symptoms rapidly resolved, but visual loss remained unchanged. CONCLUSION: The case presented here proves that GCA with typical related visual loss (AION) is possible even when both ESR and CRP are in the normal range. Therefore, in the presence of typical symptoms, the clinician must not rely solely on laboratory testing, but start steroid therapy immediately and order a temporal artery biopsy. PMID- 18386042 TI - Expression pattern of serine protease inhibitor kazal type 3 (Spink3) during mouse embryonic development. AB - Recent evidence shows that the serine protease inhibitor Kazal type 3 (Spink3) has more diverse functions than expected. To gain insight into its function, we analyzed the spatiotemporal expression profile of Spink3, using in situ hybridization (ISH) and a Spink3+/lacZ knock-in mouse, in which lacZ was inserted into the Spink3 locus. Spink3lacZ expression was first observed in the foregut, midgut, hindgut and the forebrain/midbrain junction region at 9.5 days post coitus (dpc). In the pancreas, Spink3 mRNA was detected at 11.5 dpc, before formation of the typical shape of the exocrine structure of the pancreas. Acinar cell expression was clearly identified by 13.5 dpc. After differentiation of the intestinal tract, Spink3lacZ expression was observed in the large intestine at 11.5 dpc, followed by expression in the small intestine at 13.5 dpc, before appearance of intestinal digestive enzymes. Spink3 mRNA and Spink3lacZ activity were also detected in other tissues, including the mesonephric tubules and the urogenital ridge at 11.5 dpc, the genital swelling at 13.5 dpc, the ductus epididymis at 17.5 dpc, and the seminal vesicle at 8 weeks. These data suggest that Spink3 may play important roles in proliferation and/or differentiation of various cell types during development. PMID- 18386044 TI - The art of cellular communication: tunneling nanotubes bridge the divide. AB - The ability of cells to receive, process, and respond to information is essential for a variety of biological processes. This is true for the simplest single cell entity as it is for the highly specialized cells of multicellular organisms. In the latter, most cells do not exist as independent units, but are organized into specialized tissues. Within these functional assemblies, cells communicate with each other in different ways to coordinate physiological processes. Recently, a new type of cell-to-cell communication was discovered, based on de novo formation of membranous nanotubes between cells. These F-actin-rich structures, referred to as tunneling nanotubes (TNT), were shown to mediate membrane continuity between connected cells and facilitate the intercellular transport of various cellular components. The subsequent identification of TNT-like structures in numerous cell types revealed some structural diversity. At the same time it emerged that the direct transfer of cargo between cells is a common functional property, suggesting a general role of TNT-like structures in selective, long-range cell-to cell communication. Due to the growing number of documented thin and long cell protrusions in tissue implicated in cell-to-cell signaling, it is intriguing to speculate that TNT-like structures also exist in vivo and participate in important physiological processes. PMID- 18386045 TI - Occupational class inequalities in health across employment sectors: the contribution of working conditions. AB - OBJECTIVE: While health inequalities among employees are well documented, their variation and determinants among employee subpopulations are poorly understood. We examined variations in occupational class inequalities in health within four employment sectors and the contribution of working conditions to these inequalities. METHODS: Cross-sectional data from the Helsinki Health Study in 2000-2002 were used. Each year, employees of the City of Helsinki, aged 40-60 years, received a mailed questionnaire (n = 8,960, 80% women, overall response rate for 3 years 67%). The outcome was physical health functioning measured by the overall physical component summary of SF-36. The socioeconomic indicator was occupational social class. Employment sectors studied were health care, education, social welfare and administration (n = 6,557). Physical and mental workload, and job demands and job control were explanatory factors. Inequality indices from logistic regression analysis were calculated. RESULTS: Occupational class inequalities in physical health functioning were slightly larger in education (1.47) than in the other sectors (1.43-1.40). Physical workload explained 95% of inequalities in social welfare and 32-36% in the other sectors. Job control also partly explained health inequalities. However, adjusting for mental workload and job demands resulted in larger health inequalities. CONCLUSION: Inequalities in physical health functioning were found within each employment sector, with minor variation in their magnitude. Physical workload was the main explanation for these inequalities, but its contribution varied between the sectors. In contrast, considering psychosocial working conditions led to wider inequalities. Improving physical working conditions among the lower occupational classes would help reduce health inequalities within different employment sectors. PMID- 18386046 TI - Effect of an EBM course in combination with case method learning sessions: an RCT on professional performance, job satisfaction, and self-efficacy of occupational physicians. AB - OBJECTIVE: An intervention existing of an evidence-based medicine (EBM) course in combination with case method learning sessions (CMLSs) was designed to enhance the professional performance, self-efficacy and job satisfaction of occupational physicians. METHODS: A cluster randomized controlled trial was set up and data were collected through questionnaires at baseline (T0), directly after the intervention (T1) and 7 months after baseline (T2). The data of the intervention group [T0 (n = 49), T1 (n = 31), T2 (n = 29)] and control group [T0 (n = 49), T1 (n = 28), T2 (n = 28)] were analysed in mixed model analyses. Mean scores of the perceived value of the CMLS were calculated in the intervention group. RESULTS: The overall effect of the intervention over time comparing the intervention with the control group was statistically significant for professional performance (p < 0.001). Job satisfaction and self-efficacy changes were small and not statistically significant between the groups. The perceived value of the CMLS to gain new insights and to improve the quality of their performance increased with the number of sessions followed. CONCLUSION: An EBM course in combination with case method learning sessions is perceived as valuable and offers evidence to enhance the professional performance of occupational physicians. However, it does not seem to influence their self-efficacy and job satisfaction. PMID- 18386047 TI - HGF suppresses the production of collagen type III and alpha-SMA induced by TGF beta1 in healing fibroblasts. AB - The aim of this study was to examine the effectiveness of HGF in blocking TGF beta1-induced collagen III and alpha-smooth muscle actin (alpha-SMA) production in rat healing fibroblasts, fibroblasts were obtained from healing medial collateral ligament (MCL) injury. Cell culture was supplemented with 5 ng/ml of TGF-beta1 along with increasing doses of HGF (10-40 ng/ml). The productions of collagen III in supernatants culture were assayed by enzyme-linked immunosorbent assay. Expression of alpha-SMA was assessed by Western blot. Treatment with TGF beta1 significantly stimulated collagen III and alpha-SMA production in healing fibroblasts. Remarkably, the addition of HGF reduced productions of all components induced by TGF-beta1 in a dose-dependent manner. This study shows that HGF antagonizes the action of TGF-beta1 effectively in cultured healing MCL injury fibroblasts. The results provide a cellular and molecular basis for HGF's acting as a therapeutic agent for MCL scar formation and poor healing. PMID- 18386048 TI - Is it helpful to study a commercial multi-nutrient supplement on exercise performance? PMID- 18386043 TI - The desmosome and pemphigus. AB - Desmosomes are patch-like intercellular adhering junctions ("maculae adherentes"), which, in concert with the related adherens junctions, provide the mechanical strength to intercellular adhesion. Therefore, it is not surprising that desmosomes are abundant in tissues subjected to significant mechanical stress such as stratified epithelia and myocardium. Desmosomal adhesion is based on the Ca(2+)-dependent, homo- and heterophilic transinteraction of cadherin-type adhesion molecules. Desmosomal cadherins are anchored to the intermediate filament cytoskeleton by adaptor proteins of the armadillo and plakin families. Desmosomes are dynamic structures subjected to regulation and are therefore targets of signalling pathways, which control their molecular composition and adhesive properties. Moreover, evidence is emerging that desmosomal components themselves take part in outside-in signalling under physiologic and pathologic conditions. Disturbed desmosomal adhesion contributes to the pathogenesis of a number of diseases such as pemphigus, which is caused by autoantibodies against desmosomal cadherins. Beside pemphigus, desmosome-associated diseases are caused by other mechanisms such as genetic defects or bacterial toxins. Because most of these diseases affect the skin, desmosomes are interesting not only for cell biologists who are inspired by their complex structure and molecular composition, but also for clinical physicians who are confronted with patients suffering from severe blistering skin diseases such as pemphigus. To develop disease-specific therapeutic approaches, more insights into the molecular composition and regulation of desmosomes are required. PMID- 18386049 TI - Characteristics of fast voluntary and electrically evoked isometric knee extensions during 56 days of bed rest with and without exercise countermeasure. AB - The contractile characteristics of fast voluntary and electrically evoked unilateral isometric knee extensions were followed in 16 healthy men during 56 days of horizontal bed rest and assessed at bed rest days 4, 7, 10, 17, 24, 38 and 56. Subjects were randomized to either an inactive control group (Ctrl, n = 8) or a resistive vibration exercise countermeasure group (RVE, n = 8). No changes were observed in neural activation, indicated by the amplitude of the surface electromyogram, or the initial rate of voluntary torque development in either group during bed rest. In contrast, for Ctrl, the force oscillation amplitude at 10 Hz stimulation increased by 48% (P < 0.01), the time to reach peak torque at 300 Hz stimulation decreased by 7% (P < 0.01), and the half relaxation time at 150 Hz stimulation tended to be slightly reduced by 3% (P = 0.056) after 56 days of bed rest. No changes were observed for RVE. Torque production at 10 Hz stimulation relative to maximal (150 Hz) stimulation was increased after bed rest for both Ctrl (15%; P < 0.05) and RVE (41%; P < 0.05). In conclusion, bed rest without exercise countermeasure resulted in intrinsic speed properties of a faster knee extensor group, which may have partly contributed to the preserved ability to perform fast voluntary contractions. The changes in intrinsic contractile properties were prevented by resistive vibration exercise, and voluntary motor performance remained unaltered for RVE subjects as well. PMID- 18386050 TI - Modulation of troponin C affinity for the thin filament by different cross-bridge states in skinned skeletal muscle fibers. AB - In vertebrate skeletal muscle, the C-domain of troponin C (TnC) serves as an anchor; the N-domain regulates the position of troponin-tropomyosin on the thin filament after changes in intracellular Ca2+. Another type of thin-filament regulation is provided by cross-bridges. In this study, we use skinned fibers reconstituted with chicken recombinant TnC (rTnC) to examine TnC-thin filament affinity when cross-bridges containing different ligands are formed. Dissociation and equilibrium binding of apo-TnC (i.e., lacking divalent cations) under different conditions were monitored by a standard test for maximum tension (P (o)). After 10 min in low-Mg2+ relaxing solution, rTnC dissociation (i.e., tension loss) was 80% vs only 45% in rigor. In rigor, adding myosin subfragment 1 (S1) reduced dissociation approximately twofold, whereas stretching to reduce filament overlap increased dissociation to nearly the value for relaxed fibers. Dissociation of rTnC after addition of Pi or MgADP to form A.M.Pi or A.M.ADP cross-bridges was significantly greater than with rigor (A.M) bridges. The increase in P (o) during equilibration with different concentrations of rTnC showed that the affinity for rTnC binding to the thin filament increased progressively with stronger cross-bridges: rTnC concentrations for half-maximal reconstitution (K (0.5)) were 8.1, 3.7, 2.9, and 1.1 microM for A + M.ADP.Pi, A.M.Pi, A.M, and A.M + S1. Cross-bridges containing MgADP(-) (A.M.ADP) were also less effective than rigor bridges in promoting rTnC binding. We conclude that cross-bridge state and number both modulate TnC affinity for the thin filament and that the TnC C-domain is a central element in this pathway. PMID- 18386051 TI - The G604S-hERG mutation alters the biophysical properties and exerts a dominant negative effect on expression of hERG channels in HEK293 cells. AB - We have recently identified a missense mutation, G604S, in the human ether-a-go go related gene (hERG) that results in a malignant phenotype in a full pedigree of a Chinese congenital long QT syndrome (LQTS) family. The present study characterized the pathophysiological consequences of the mutation at the cellular level. Mutant G604S-hERG channels were expressed in HEK293 cells using a lipofectamine method. hERG currents were recorded using the voltage clamp technique. The expression of hERG protein was detected by Western blotting, and the subcellular location of hERG channels in cell was analyzed by confocal microscopy. We found that the G604S mutation did not lead to any expression of detectable currents, which was consistent with Western blotting analysis that the G604S-hERG mutation only expressed a band at 135 kDa. When coexpressed with wild type (WT)-hERG, G604S-hERG exhibited strong dominant-negative current suppression resulting in decreased current density and altered gating properties of the WT hERG channel, as well as interference with the trafficking of WT-hERG channel protein. In addition, confocal microscopy demonstrated that G604S-hERG subunits could be inserted into the cell membrane when forming heteromultimeric channels with WT-hERG channel subunits. Our results suggest that G604S mutation causes a loss of function in hERG through a strong dominant-negative effect on WT-hERG channel function that caused by impaired trafficking of WT-hERG channels, and further accentuates this suppression by forming heteromultimeric functional channels with WT-hERG subunits. PMID- 18386052 TI - Wide distribution of external local sign in the normal population. AB - The extent of the apparent visual field was determined for a group of 78 naive visual observers. We find that there exists a minority (less than 10%) that is essentially veridical, but that the majority of the population experiences an apparent visual field of only about 90 degrees, thus much narrower than the dioptrics of the eye would suggest (a little over 180 degrees). This is in good accordance with available (albeit mainly anecdotal) evidence, though formal data have been lacking thus far. The finding is discussed in the context of metrical calibration of the topological structure of the visual field, an aspect of "local sign". PMID- 18386054 TI - Trisomy of chromosome 6 in Merkel cell carcinoma within lymph nodes. AB - Merkel cell carcinoma (MCC) of the skin is a neuroendocrine tumor with characteristic histological and immunohistochemical features. Among various cytogenetic changes, trisomy of chromosome 6 has been reported in 47% of cases using in situ hybridization. Primary tumors, morphologically and immunohistochemically identical to MCCs of the skin, have been described in other organs, including lymph nodes. Here, a cytogenetic study of four cases of MCC of lymph nodes is presented. Four cases of primary MCCs of lymph nodes and ten cases of cutaneous MCCs were studied for chromosome 6 using fluorescent in situ hybridization (FISH). All cases showed typical features of MCC both at hematoxylin and eosin (H&E) and immunohistochemistry. FISH showed trisomy 6 in two out of the four cases ofMCCs of lymph node as well as in 6 out 10 cases of MCCs of skin. Lymph nodal and cutaneous MCCs share same histological and immunohistochemical features, as well as same cytogenetic alteration for chromosome 6. It seems that there are more similarities than differences between cutaneous and lymph nodal MCCs. Whether lymph nodal MCCs are primary tumors or metastases from regressed skin lesions is still questionable, although several findings indicate a primary origin. PMID- 18386053 TI - Sarcoma of follicular dendritic cells with features of sinus lining cells--a new subtype of reticulum cell sarcoma? AB - Dendritic cell neoplasms of the World Health Organization classification comprise Langerhans cell histiocytosis, Langerhans cell sarcoma, interdigitating dendritic cell sarcoma, follicular dendritic cell sarcoma, and dendritic cell sarcoma, not otherwise specified. Several studies based on immunohistochemical and ultrastructural analysis tried to further clarify the origin of these neoplasms which are thought to derive from mesenchymal or bone marrow precursors. Lymphatic vessel endothelium hyaluronan receptor-1 (LYVE-1) was recently described as a marker for lymphatic endothelium which is expressed on normal liver blood sinusoid lining cells, spleen endothelium, activated tissue macrophages, blood vessels in the lung, endothelial cells of lymphatic sinuses, and in fibroblastic reticular cells in lymph nodes. We present a case of LYVE-1-positive reticulum cell neoplasm in an axillary lymph node. To the best of our knowledge, there has been no report about LYVE-1 expression in histiocytic or dendritic cell neoplasms so far. Due to the assumed specificity of this antibody, we propose designation of this reticulum cell sarcoma as lymphatic sinus lining cell sarcoma which might finally represent another subtype of reticulum cell sarcomas. PMID- 18386055 TI - IIICS de novo glycosylated fibronectin as a marker for invasiveness in urothelial carcinoma of the urinary bladder (UBC). AB - PURPOSE: The urothelial carcinoma is the most frequent malignancy of the urinary bladder (UBC). The transition into invasive growth is accompanied by several histological changes including an oncofoetal reorganization of the extracellular matrix. Recently, the occurrence of oncofoetal fibronectin with an O-linked glycosylation in the IIICS region (oncf Fn) was shown to be present in urine from UBC patients and was recommended as a tumour marker. Until now there are no data available regarding the source and distribution of oncf Fn in UBC and its value for the assessment of invasiveness. METHODS: oncf Fn was analysed in noninvasive and invasive UBC using immunohistochemistry and western blot. Additionally, the mRNA expression of the IIICS splicing region was evaluated by quantitative real time RT-PCR. RESULTS: Immunohistochemical results reveal a highly significant correlation of oncf Fn to invasiveness. Papillary tumours regularly show no positivity. In western blot, invasive UBC show a strongly increased amount of the 250 kDa oncf Fn. Additionally, several smaller bands could be shown suggesting a proteolytic processing of Fn. The mRNA of the IIICS region shows a 21.5-fold increase in invasive UBC compared with noninvasive carcinomas. CONCLUSIONS: In summary, immunohistochemistry of oncf Fn is a valuable histological marker for invasiveness of urothelial carcinoma of the urinary bladder. The restricted and invasion-associated tissue distribution of immunoreactivity enables to monitor the recurrence of invasive UBC by a quantitative evaluation of IIICS O-linked glycosylated Fn in urine. PMID- 18386056 TI - The role of omentectomy during the surgical staging in patients with clinical stage I endometrioid adenocarcinoma. AB - OBJECTIVE: The aim of this study was to evaluate whether omentectomy should be a routine part of staging surgery in endometrioid adenocarcinoma. METHODS: A retrospective study was performed on 65 patients who were primarily treated by total abdominal hysterectomy, salpingo-oophorectomy, bilateral pelvic and para aortic lymphadenectomy, infracolic omentectomy and peritoneal cytology for clinical stage I endometrial carcinoma between January 2002 and December 2005. Data on 65 patients who had been diagnosed with clinical stage I endometrial carcinoma were reviewed. Associations in the data obtained, pelvic and para aortic lymph node status, depth of myometrial invasion, grade, vascular invasion, adnexal involvement, positive peritoneal cytology, lymph node metastasis, cervical stromal invasion, and tumor size, were investigated. The Chi-square (chi(2)) test was used for statistical analysis. Multivariate analysis was performed with logistic regression analyses. RESULTS: Four (6.2%) of 65 patients had omental metastasis, which was microscopic in two patients. As for extrauterine spread, the positivity rate of lymph node metastases was 10/65 (15.38%), peritoneal cytology was 7/65 (10.76%), and adnexal metastases was 10/65 (15.38%). Of those patients with omental metastasis, 2/10(20%) had positive nodes, 2/10(20%) had adnexal metastases, and 3/7(42.8%) had positive peritoneal cytologic findings. These four patients with omental metastasis had significantly higher rates of positive cytology (P = 0.003). Multivariate analysis revealed omental metastasis (P = 0.002; OR 46.5, CI 95% 3.899-554.575) to be significantly associated with positive peritoneal cytology CONCLUSIONS: We conclude that despite the presence of normal-appearing omentum, omentectomy should be performed as a component of surgical staging in the presence of positive peritoneal cytology. PMID- 18386057 TI - Usefulness of limb salvage surgery for bone and soft tissue sarcomas of the distal lower leg. AB - PURPOSE: The usefulness of limb salvage surgery for distal lower leg sarcoma remains controversial. We analyzed the long-term prognosis, limb function, and complications after limb salvage treatment of patients with distal lower leg sarcoma. METHODS: Ten patients treated with limb salvage surgery for primary distal lower leg sarcoma were retrospectively reviewed. The median follow-up period after the first operation was 9.0 years. We performed three types of reconstructive techniques for the skeletal defect after a wide resection, including (1) arthrodesis with a combination of autograft and intraoperative autoclaved tumor bone graft in two patients, or allograft in one patient, (2) ankle joint preserving surgery using intraoperative extracorporeal irradiated tumor bone graft in five patients, and (3) prosthesis in two patients. RESULTS: The overall survival rate was 80%. The 5-year disease-free survival was 80%. The rate of limb preservation at the final follow-up was 90%. The mean functional score according to the scoring system of the Musculoskeletal Tumor Society was 88% at the final follow-up. Postoperative complications occurred in seven patients. Skin trouble occurred in three patients, infectious non-union in one patient, fracture in three patients, and loosening of prosthesis in one patient. CONCLUSIONS: Despite the high rate of complications, patients treated with limb salvage surgery for the distal lower leg sarcoma revealed excellent final functional results without impairing the oncologic results. Limb salvage surgery is therefore considered to be an effective treatment option for distal lower leg sarcoma when adequate informed consent can be obtained from the patient. PMID- 18386058 TI - Alterations of RB1 gene in embryonal and alveolar rhabdomyosarcoma: special reference to utility of pRB immunoreactivity in differential diagnosis of rhabdomyosarcoma subtype. AB - PURPOSE: Rhabdomyosarcoma (RMS), which is the most common pediatric soft tissue sarcoma, is classified into two major histologic subtypes, embryonal RMS (ERMS) and alveolar RMS (ARMS). RMS is occasionally reported to be the second neoplasm of hereditary retinoblastoma. Osteosarcoma is known as the most common second neoplasm of hereditary retinoblastoma, and tumorigenesis of osteosarcoma has been proven in previous studies to be related to the RB gene (RB1) alteration. Therefore, there might be a correlation between the tumorigenesis of RMS and RB1 alteration. METHODS: We examined the RB protein (pRB) expression and RB1 alteration such as allelic imbalance (gain or loss) and homozygous deletion, using immunohistochemistry, microsatellite makers, and quantitative real-time PCR in 57 sporadic RMS. RESULTS: Allelic imbalance was more frequently detected in ERMS (13/27), than in ARMS (3/20) (P = 0.04). Homozygous deletion on the protein binding pocket domain of RB1 was found in 6 of 27 ERMS and in 2 of 20 ARMS (P = 0.24). Furthermore, immunohistochemical pRB labeling indexes (LI) in 31 ERMS (median value, 31%) were significantly reduced in comparison with those observed in 26 ARMS (median value, 85%) (P < 0.0001). CONCLUSIONS: Our results support the assertion that tumorigenesis of RMS may be associated with RB1 alteration especially in ERMS, as previously reported for osteosarcoma. As for the RB pathway, each subtype of RMS may have a different tumorigenesis. In addition, immunohistochemical pRB LI may have the potential to be a useful ancillary tool in the differential diagnosis of RMS subtypes. PMID- 18386059 TI - FISH analysis of chromosomes 3 and 6 on fine needle aspiration biopsy samples identifies distinct subgroups of uveal melanomas. AB - PURPOSE: Circumstantial evidence suggests that development of uveal melanoma may be associated to two different pathogenetic pathways, either loss of chromosome 3 or extra copies of 6p (+6p). Chromosome 3 monosomy (-3) is detected in approximately half of uveal melanomas, and is strongly linked to metastatic disease, whereas +6p accounts for one-fourth of uveal melanomas with no clear clinical correlations. The aim of our study was to verify if the analysis of chromosomes 3 and 6 was able to distinguish two groups of patients for translating this approach in the clinical practise as prognostic tool. METHODS: Fluorescence in situ hybridisation (FISH) with probes for chromosome 3, 6p and 6q was used to analyze cytological material obtained by fine needle aspiration biopsy (FNAB) from 28 primary uveal melanomas, just before brachytherapy. RESULTS: Abnormalities affecting 6p and 6q were found in 14 tumors (50%), and -3 in 16 cases (57%). Interestingly, -3 and +6p were mutually exclusive in 23 cases (82%), whereas in two cases only (7%) they coexisted. In particular, +6p alone was present in 9 lesions (32%), -3 was the sole aberration in 11 cases (39%), and concomitant -3 and -6q in 3 other cases (11%). CONCLUSIONS: Although the patient cohort is limited, our findings confirm the hypothesis of a bifurcated pathogenetic model of uveal melanoma. Moreover, our results suggest that investigation of both markers on FNAB samples obtained in vivo could provide a clearer clinical picture of genetic lesions when no histological material is available for prognostic evaluation. PMID- 18386060 TI - Allelic loss on chromosome 5q34 is associated with poor prognosis in hepatocellular carcinoma. AB - PURPOSE: To identify and characterize novel genetic alterations in hepatocellular carcinoma (HCC). METHODS: DNA was extracted from 29 HCC and corresponding normal tissues and amplified with 59 different 10-base arbitrary primers. A 550 bp DNA fragment amplified using primer Q-9 and which was present in 19 of 29 cases (66%) was cloned, sequenced, and compared with known nucleotide sequences deposited in Genome database, and quantified by real-time PCR. RESULTS: DNA alterations were found on chromosomes 5q34, 6p25.2 and 8q12.1 in 11 of 29 cases (38%), 7 of 29 cases (24%), and 12 of 29 cases (41%), respectively. Multivariate analysis showed that the allelic loss on chromosome 5q34 was an independent prognostic factor for poor survival of HCC patients, with the median survival time of 19 weeks for allelic loss versus 109 weeks for no allelic loss (P = 0.001). CONCLUSIONS: This study indicates that allelic loss on chromosome 5q34 may be involved in the development of HCC and could be used as a prognostic indicator in HCC patients. PMID- 18386061 TI - Friendship in research: the Japan-Korea Symposia on Cancer and Ageing. PMID- 18386062 TI - Biometrical and genetical characterization of large Babesia ovis in Iran. AB - One species of Babesia was identified on the blood smear of 20 different naturally infected sheep in the Northwest of Iran. It was polymorphic, including double pyriform with acute or obtuse angle, single pyriform, and ring form. The size of typical paired pyriforms with acute angle was 2.7 x 0.4 microm (n=10) and with obtuse angle was 3.5 x 0.6 microm (n=10). Although the morphological and biometrical parameters resembled the Babesia motasi, the results of seminested polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism using primers specific for small subunit of 18S rRNA confirmed this species as Babesia ovis. Furthermore, the sequence analysis of hypervariable region of small subunit of 18S rRNA revealed the corresponding sequences for B. ovis as well. Experimental infection of healthy lambs with the morphological larger B. ovis showed a milder clinical signs compared to the small one. PMID- 18386063 TI - Efficacy of imidacloprid 10%/moxidectin 1% (Advocate/Advantage Multi) against fleas (Ctenocephalides felis felis) on ferrets (Mustela putorius furo). AB - Ferrets (Mustela putorius furo) are becoming increasingly popular as pets and are kept in households with other pet animals such as dogs and cats, from which they may catch flea infestations with the predominant flea species Ctenocephalides felis. In this study, the efficacy of imidacloprid/moxidectin spot-on (Advocate/Advantage Multi) was investigated in the therapy and prevention of flea infestation in ferrets. Sixteen adult ferrets of varying weights and ages and of both sexes were included. Ferrets were infested with 50 fleas each on days -7, 1, 7, 14, 21, and 28. On study day -6, the animals were randomized into two groups based on body weight and flea count. Each ferret allocated to group 1 was treated with 0.4 ml of imidacloprid 10%/moxidectin 1% (Advocate/Advantage Multi). The ferrets in group 2 remained untreated. Flea counts were performed by combing 24 to 48 h after infestation. The primary efficacy criterion was the reduction in the number of fleas at each time point post treatment compared to the ferrets in the untreated control group. On day 1, the therapeutic efficacy was 100%. The preventative efficacy was 100% at 1 and 2 weeks post treatment, and it was >97% and >90% at 3 and 4 weeks post treatment. No local or systemic side effects were observed in any of the ferrets treated. PMID- 18386065 TI - A novel tissue-slice culture model for non-malignant human prostate. AB - A novel tissue culture system was established for modeling the non-neoplastic human prostate in vitro. Precision-cut prostate slices were cultivated in culture plates with a gas-permeable base in a novel serum-free mixture. Cultivated specimens was evaluated by an immunohistochemical analysis of cytokeratins 18 and 14, androgen receptor (AR), prostate specific antigen (PSA), prostate acid phosphatase (PAP), and the endothelial cell marker von Willebrand factor. Epithelial viability in the presence and absence of dihydrotestosterone (DHT) was also assessed. Satisfactory maintenance of glandular cytoarchitecture was observed in the presence of DHT with approximately half of the glands displaying a columnar or cuboidal phenotype and an intact layer of basal cells. In the absence of DHT, the corresponding percentage was significantly lower. The occurrence of involutive changes and epithelial cell death was significantly higher in the absence of DHT. Glandular and stromal cells maintained their capacity to express AR. PSA and PAP were expressed throughout the culture period, albeit at a lower level than in uncultured tissue. The viability of endothelial cells differed markedly between individual samples. During culture, the tissue slices became covered with epithelial cells originating from glands that were cut open during tissue slicing. This cell layer consisted of a stratified basal compartment overlaid by cells with a luminal phenotype. The present culture system provides a novel in vitro setting in which to study normal human prostate biology and pathobiology and may help to obviate problems related to the use of established cancer cell lines and animal models. PMID- 18386064 TI - Exploring the transcriptome of the burrowing nematode Radopholus similis. AB - Radopholus similis is an important nematode pest on fruit crops in the tropics. Unraveling the transcriptome of this migratory plant-parasitic nematode can provide insight in the parasitism process and lead to more efficient control measures. For the first high throughput molecular characterization of this devastating nematode, 5,853 expressed sequence tags from a mixed stage population were generated. Adding 1,154 tags from the EST division of GenBank for subsequent analysis, resulted in a total of 7,007 ESTs, which represent approximately 3,200 genes. The mean G + C content of the nucleotides at the third codon position (GC3%) was calculated to be as high as 64.8%, the highest for nematodes reported to date. BLAST-searches resulted in about 70% of the clustered ESTs having homology to (DNA and protein) sequences from the GenBank database, whereas one third of them did not match to any known sequence. Roughly 40% of these latter sequences are predicted to be coding, representing putative novel protein coding genes. Functional annotation of the sequences by GO annotation revealed the abundance of genes involved in reproduction and development, which reflects the nematode population biology. Genes with a role in the parasitism process are identified, as well as genes essential for nematode survival, providing information useful for parasite control. No evidence was found for the presence of trans-spliced leader sequences commonly occurring in nematodes, despite the use of various approaches. In conclusion, we found three different sources for the EST sequences: the majority has a nuclear origin, approximately 1% of the EST sequences are derived from the mitochondrial transcriptome, and interestingly, 1% of the tags are with high probability derived from Wolbachia, providing the first molecular indication for the presence of this endosymbiont in a plant-parasitic nematode. PMID- 18386066 TI - Establishment of a preadipocyte cell line derived from mature adipocytes of GFP transgenic mice and formation of adipose tissue. AB - We established a preadipocyte cell line from mature adipocytes obtained from subcutaneous fat tissue of green fluorescent protein (GFP) transgenic mice. The floating top layer, containing mature adipocytes, was isolated from subcutaneous fat tissue by collagenase digestion and filtration. Fluorescence-activated cell sorting and microscopic analysis revealed that the floating cell fraction comprised a highly homogeneous adipocyte population with no adipose stromal vascular cells. Isolated mature adipocytes dedifferentiated into fibroblast-like cells and actively proliferated in ceiling culture. In vitro studies showed that the cells could redifferentiate into mature adipocytes in an identical way to 3T3 L1 preadipocytes. No changes in the differentiation pattern were observed during the propagation of our cells. They were successfully maintained and differentiated for at least 22 passages. We named these cells dedifferentiated fat (DFAT-GFP) cells. When DFAT-GFP cells were implanted subcutaneously into C57BL/6N mice, they developed highly vascularized fat pads that morphologically resembled normal subcutaneous adipose tissue and consisted of GFP-positive cells; however, implanted 3T3-L1 cells did not have such an effect on the mice. We conclude that DFAT-GFP cells provide a model that should enable us to study the mechanisms of adipocyte differentiation and adipose tissue formation in vivo and in vitro. PMID- 18386067 TI - Correlated evolution of fig size and color supports the dispersal syndromes hypothesis. AB - The influence of seed dispersers on the evolution of fruit traits remains controversial, largely because most studies have failed to account for phylogeny and or have focused on conservative taxonomic levels. Under the hypothesis that fruit traits have evolved in response to different sets of selective pressures by disparate types of seed dispersers (the dispersal syndromes hypothesis), we test for two dispersal syndromes, defined as groups of fruit traits that appear together more often than expected by chance. (1) Bird syndrome fruits are brightly colored and small, because birds have acute color vision, and commonly swallow fruits whole. (2) Mammal syndrome fruits are dull-colored and larger on average than bird syndrome fruits, because mammals do not rely heavily on visual cues for finding fruits, and can eat fruits piecemeal. If, instead, phylogenetic inertia determines the co-occurrence of fruit size and color, we will observe that specific combinations of size and color evolved in a small number of ancestral species. We performed a comparative analysis of fruit traits for 64 species of Ficus (Moraceae), based on a phylogeny we constructed using nuclear ribosomal DNA. Using a concentrated changes test and assuming fruit color is an independent variable, we found that small-sized fruits evolve on branches with red and purple figs, as predicted by the dispersal syndromes hypothesis. When using diameter as the independent variable, results vary with the combination of algorithms used, which is discussed in detail. A likelihood ratio test confirms the pattern found with the concentrated changes test using color as the independent variable. These results support the dispersal syndromes hypothesis. PMID- 18386068 TI - The role of density-dependent individual growth in the persistence of freshwater salmonid populations. AB - Theoretical and empirical models of populations dynamics have paid little attention to the implications of density-dependent individual growth on the persistence and regulation of small freshwater salmonid populations. We have therefore designed a study aimed at testing our hypothesis that density-dependent individual growth is a process that enhances population recovery and reduces extinction risk in salmonid populations in a variable environment subject to disturbance events. This hypothesis was tested in two newly introduced marble trout (Salmo marmoratus) populations living in Slovenian streams (Zakojska and Gorska) subject to severe autumn floods. We developed a discrete-time stochastic individual-based model of population dynamics for each population with demographic parameters and compensatory responses tightly calibrated on data from individually tagged marble trout. The occurrence of severe flood events causing population collapses was explicitly accounted for in the model. We used the model in a population viability analysis setting to estimate the quasi-extinction risk and demographic indexes of the two marble trout populations when individual growth was density-dependent. We ran a set of simulations in which the effect of floods on population abundance was explicitly accounted for and another set of simulations in which flood events were not included in the model. These simulation results were compared with those of scenarios in which individual growth was modelled with density-independent Von Bertalanffy growth curves. Our results show how density-dependent individual growth may confer remarkable resilience to marble trout populations in case of major flood events. The resilience to flood events shown by the simulation results can be explained by the increase in size-dependent fecundity as a consequence of the drop in population size after a severe flood, which allows the population to quickly recover to the pre-event conditions. Our results suggest that density-dependent individual growth plays a potentially powerful role in the persistence of freshwater salmonids living in streams subject to recurrent yet unpredictable flood events. PMID- 18386069 TI - Outcome after renal transplantation. Part II: quality of life and psychosocial adjustment. AB - Knowledge of health-related quality of life (QOL) and psychosocial adjustment (PA) in children after renal transplantation (RTPL) is limited. QOL and PA were evaluated by standardized tests in patients after RTPL. Thirty-seven children of median age 14.5 years (range 6.5-17 years) were investigated a mean 4.5 years (range 0.5-12.8 years) after RTPL. Child- and parent-rated QOL was evaluated with the Child Quality of life Questionnaire of The Netherlands Organization for Applied Scientific Research Academical Medical Centre (TNO-AZL). PA was assessed by the Child Behaviour Checklist (CBCL) providing parental reports of a child's behaviour. In patients' self-ratings, the QOL dimension physical complaints (P < 0.0005) scored significantly better than that of healthy controls, whereas the dimension positive emotional functioning was impaired (P = 0.02). Parents rated motor functioning (P = 0.002), autonomy (P = 0.01), cognition (P = 0.04) and positive emotions (P < 0.0005) as significantly impaired. Parents also assessed PA significantly (P = 0.02) impaired with regard to internalizing behaviour. Dialysis duration, young age at RTPL, living-related donation, steroid treatment, adverse family relationships and maternal distress had a significantly negative impact on QOL and PA (P < 0.05). Patients rated QOL higher than did healthy controls. Parents evaluated their children's QOL and PA more pessimistically than did the patients themselves. Both illness-related variables and family environment played an important role. PMID- 18386070 TI - Atypical presentation of distal renal tubular acidosis in two siblings. AB - Primary distal renal tubular acidosis (dRTA) is an inherited disease characterized by the inability of the distal tubule to lower urine pH <5.50 during systemic acidosis. We report two male siblings who presented with severe hyperchloremic metabolic acidosis, high urinary pH, nephrocalcinosis, growth retardation, sensorineural hearing loss, and hypokalemic paralysis. Laboratory investigations revealed proximal tubular dysfunction (low molecular weight proteinuria, generalized hyperaminoaciduria, hypophosphatemia with hyperphosphaturia, and hypouricemia with hyperuricosuria). There was significant hyperoxaluria and laboratory evidence for mild rhabdomyolysis. Under potassium and alkali therapy, proximal tubular abnormalities, muscular enzymes, and oxaluria normalized. A homozygous mutation in the ATP6V1B1 gene, which is responsible for dRTA with early hearing loss, was detected in both siblings. In conclusion, proximal tubular dysfunction and hyperoxaluria may be found in children with dRTA and are reversible under appropriate therapy. PMID- 18386071 TI - Can anorexia predict patient satisfaction with quality of life in advanced cancer? AB - GOALS: Anorexia is the second most common symptom after fatigue in patients with advanced cancer. We quantified the relationship between anorexia and patient satisfaction with quality of life (QoL) in advanced cancer. MATERIALS AND METHODS: A case series of 954 cancer patients treated at the Cancer Treatment Centers of America. Anorexia was measured using the QLQ-C30 loss of appetite/anorexia subscale. Patient satisfaction with QoL was measured using the Ferrans and Powers Quality of Life Index (QLI). The relationship between anorexia and QLI was evaluated using multivariate analysis of variance and multiple regression analysis. MAIN RESULTS: Of 954 patients, 579 were females and 375 males with the median age at presentation of 56 years. Of these patients, 66% had failed prior treatment. Most common cancers were breast (26%), colorectal (19%), and lung (16%). After controlling for the effects of age, treatment history, and other QLQ-C30 symptom subscales, every 10 unit increase in anorexia was statistically significantly associated with 0.18 unit decline in QLI health and physical function score. CONCLUSION: We found that anorexia is significantly correlated with patient satisfaction with health and physical function in advanced cancer. Future studies should evaluate the impact of integrative cancer care services on patient satisfaction with QoL. PMID- 18386073 TI - Measurement of self-advocacy in cancer patients and survivors. AB - BACKGROUND: The Patient Self-Advocacy Scale, originally developed for use in HIV/AIDS patients, was adapted for use in cancer patients and survivors. The scale contains three dimensions: increased illness education, increased assertiveness with a doctor, and potential for mindful non-adherence. DISCUSSION AND CONCLUSION: Evidence for modest reliability and construct validity was found; however, ceiling effects exist for the Illness Education subscale, which supports the need to move beyond items regarding information seeking to perhaps include use and management of information. PMID- 18386072 TI - Causal attributions to epidemiological risk factors and their associations to later psychological adjustment among Japanese breast cancer patients. AB - GOALS: The objective of this study was to evaluate the causal attributions for breast cancer and their association with the subsequent psychological adjustment. MATERIALS AND METHODS: Sixty-three Japanese patients newly diagnosed with breast cancer were asked by an interviewer about risk factors and explanations for the etiology of breast cancer using a four-point scale. The General Health Questionnaire (GHQ-28) and the Center for Epidemiologic Studies Depression Scale (CES-D) were administered about 2 months later to measure psychological adjustment to the illness. The relationship between each causal attribution and measure of psychological adjustment was assessed with the Spearman rank correlation after adjusting for potential confounders (age, marital status, cancer stage at diagnosis, time between the day of diagnosis and follow-up, and physical symptoms at the follow-up). MAIN RESULTS: Attributions to several explanations, including "stress" and "personal characteristics," were positively associated with the GHQ-28 measures. Of the attributions to risk factors, "body size" was significantly inversely (r = -0.29) associated and "never having children" was insignificantly and highly inversely (r = -0.77) associated with the GHQ-28 measures, indicating better adjustment to the illness. On the other hand, the attribution to "tobacco" was significantly and positively associated with the GHQ-28 measures (r = 0.34), indicating that the attributions antagonized adjustment to the illness. CONCLUSION: The current study indicates that forming causal attributions influences the adjustment to the illness in Japanese breast cancer patients and attributions to certain risk factors for breast cancer may contribute to better adjustment. PMID- 18386074 TI - The religiousness as a way of coping with anxiety in women with breast cancer at different disease stages. AB - BACKGROUND: Anxiety is an unpleasant emotion affecting patients with cancer, and there are various ways of coping with it. Little is known about the relationship between the anxiety level and physical, somatic or spiritual (e.g. religiousness) factors in breast cancer patients at different stages of the disease. OBJECTIVES: The purpose of the study was to assess the intensity of anxiety at different stages of breast cancer, to define the relationship between religiousness and physical (somatic) condition and anxiety in the study subjects and to find out if religiousness is an effective coping strategy at any breast cancer stage. MATERIALS AND METHODS: The study involved 180 women aged between 28 and 77, who were qualified to one of five study groups, according to their disease stage. The following research instruments were used: the State-Trait Anxiety Inventory (STAI), Scale of Personal Religiousness, the Rotterdam Symptom Checklist as well as medical history and data from patients' medical records. RESULTS: There is a significant correlation between state anxiety and trait anxiety measured by STAI in study groups. Breast cancer stage differentiates the study groups in respect of revealed anxiety level. Somatic condition has no significant impact on the intensity of anxiety in terminal breast cancer patients. Amongst the studied variables, only religiousness is an important factor that influences the anxiety level of end-stage cancer patients. CONCLUSIONS: The obtained results allow us to conclude that: (a) religiousness is an effective factor of coping with anxiety only of the end-stage breast cancer patients; (b) cancer stage is a differentiating factor in respect of revealed anxiety level in study subjects; (c) exacerbation of somatic symptoms does not influence the anxiety level in terminal cancer patients and at disease-free period. PMID- 18386075 TI - Information and service needs for young adult cancer patients. AB - BACKGROUND: Young adulthood is a time of increased vulnerability to stress and presents young adult cancer patients and off-treatment survivors with major developmental challenges above and beyond those faced by their peers. The intent of this study was to examine supportive care needs and preferences among young adult cancer patients. METHODS: Two hundred and seventeen young adults aged 18-40 years and diagnosed with cancer between the ages of 15-35 completed an online survey. Adapted from prior research on young adult survivors of childhood cancer, questions assessed the extent to which subjects had utilized various information and supportive care services and/or expressed a desire to use those services either now or in the past. RESULTS: While young adult patients demonstrate a high demand for information and assistance regarding diet and nutrition, exercise, fertility options, complementary and alternative services and assistance with health insurance, 40-50% of patients report that these needs are unmet. More than 50% of respondents who indicated a need or desire for camp programs and retreats, counseling or guidance related to sexuality, counseling for family members, infertility treatment and adoption services, transportation assistance, child care and alcohol or drug abuse counseling also indicated that these needs were unmet. Younger respondents (18-29 years olds) were significantly more likely to indicate unmet needs for the supportive care services assessed here. CONCLUSION: Findings serve as recommendations as to when and to whom to target delivery of supportive care services. PMID- 18386076 TI - Routine symptom assessment: good for practice and good for business. PMID- 18386077 TI - Noncancer life stresses in newly diagnosed cancer. AB - GOALS OF WORK: Noncancer life stresses affect psychosocial stress processes and have an impact on quality of life (QOL) of the patients. However, investigating life stresses in cancer is a recent development. We evaluated the life stresses of newly diagnosed melanoma, breast cancer, and prostate cancer patients in detail and investigated their impact on QOL outcomes after localized cancer diagnosis. MATERIALS AND METHODS: Life change events from the previous year (negative events, positive events, total impact of events, impacts of the negative events, and impacts of loss events) and chronic ongoing life strains were measured with the Life Experience Survey and the Chronic Strains Survey in newly diagnosed patients 3 months after the diagnosis. Also, perceived symptoms and QOL were measured, and in melanoma and breast cancer, these were repeated up to 2 years later. RESULTS: Noncancer life stress was common in newly diagnosed cancer patients: Both acute and chronic life stresses were experienced by four fifths. Loss events (fateful negative events or social loss events) were reported by one-third. Many patients had a preceding chronic illness. Along with the cancer and treatment stresses, the noncancer life stresses predicted poorer QOL, particularly psychological and depressive symptoms. Different life stresses predicted slightly different domains of QOL, and depressive symptoms tended to be predicted by several kinds of life stresses. Baseline life stresses had impact also on later QOL in breast cancer. CONCLUSIONS: Noncancer life stresses are common among newly diagnosed cancer patients and have impact on QOL, and thus they should be taken into account in cancer care. Screening for noncancer life stresses may offer means to enhance QOL outcomes in cancer. PMID- 18386078 TI - Arbuscular mycorrhizal fungal communities in sub-Saharan savannas of Benin, West Africa, as affected by agricultural land use intensity and ecological zone. AB - The rapid decline of soil fertility of cultivated lands in the sub-Saharan savannas of West Africa is considered to be the main cause of the increasingly severe constraints of food production. The soils in this tropical area are highly fragile, and crop yields are limited by characteristically low levels of available phosphorus. Under such preconditions, the multiple benefits of the arbuscular mycorrhizal (AM) symbiosis are likely to play a pivotal role for maintaining natural soil fertility by enhancing plant nutrient use efficiency, plant health, and stabilization of a favorable soil structure. Thus, it is important to explore the impact of the commonly applied farming practices on the native AM fungal community. In the present study, we determined the AM fungal species composition in three ecological zones differing by an increasingly prolonged dry season from South to North, from the Southern Guinea Savanna (SG), to the Northern Guinea Savanna (NG), to the Sudan Savanna (SU). In each zone, four "natural" and four "cultivated" sites were selected. "Natural" sites were three natural forest savannas (at least 25-30 years old) and a long-term fallow (6-7 years old). "Cultivated" sites comprised a field with yam (Dioscorea spp.) established during the first year after forest clearance, a field under mixed cropping with maize (Zea mays) and peanut (Arachis hypogaea), a field under peanut, and a field under cotton (Gossypium hirsutum) which was the most intensively managed crop. Soil samples were collected towards the end of the wet season in each zone. AM fungal spores were extracted and morphologically identified. Soil subsamples were used to inoculate AM fungal trap cultures using Stylosanthes guianensis and Brachiaria humidicola as host plants to monitor AM root colonization and spore formation over 10 and 24 months, respectively. A total of 60 AM fungal species were detected, with only seven species sporulating in the trap cultures. Spore density and species richness were generally higher in the natural savannas and under yam than at the other cultivated sites and lowest under the intensively managed cotton. In the fallows, species richness was intermediate, indicating that the high richness of the natural savannas was not restored. Surprisingly, higher species richness was observed in the SU than in the SG and NG, mainly due to a high proportion of species in the Gigasporaceae, Acaulosporaceae, and Glomeraceae. We conclude that the West African savannas contain a high natural AM fungal species richness, but that this natural richness is significantly affected by the common agricultural land use practices and appears not to be quickly restored by fallow. PMID- 18386079 TI - The metal binding site of zoocin A. AB - Direct metal analysis of the bacteriolytic exoenzyme zoocin A failed to unequivocally identify a putative metal cofactor; hence, indirect experiments utilizing NMR were undertaken to settle this question. Cd(2+) as a surrogate metal ion was reconstituted into EDTA-treated, metal-free recombinant zoocin, and (113)Cd-NMR was employed to explore binding in the protein for this ion. The Cd substituted enzyme was found to have 80-85% of native streptococcolytic activity. A major (113)Cd resonance at 113.6 ppm was observed which with time split into resonances at 113.6 and 107.2 ppm. A minor (113)Cd resonance at 87.3 ppm was observed which increased in intensity with time. These Cd chemical shifts are indicative of two N atoms and two O atoms ligating directly to the metal site. On the basis of conserved amino acid residues in a homologous protein of known structure, LytM, the ligands in zoocin are tentatively assigned to H45, D49, H133, and some combination of water or buffer ions as the fourth oxygen donor in zoocin A. Comparison of the combined intensities for (113)Cd-substituted zoocin with a known quantity of another Cd-substituted protein gave Cd binding as approximately stoichiometric (1.2+/-0.2) with protein. Additional metal-removal and reconstitution experiments on the recombinant catalytic domain of zoocin implicate Zn(2+) as the metal cofactor. Therefore, the evidence supports zoocin as a single Zn(2+) ion binding metalloenzyme. PMID- 18386080 TI - Cloning, expression and physicochemical characterization of a di-heme cytochrome c (4) from the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC 125. AB - The 20-kDa di-heme cytochrome c (4) from the psycrophilic bacterium Pseudoalteromonas haloplanktis TAC 125 was cloned and expressed in Escherichia coli and investigated through UV-vis and (1)H NMR spectroscopies and protein voltammetry. The model structure was computed using the X-ray structure of Pseudomonas stutzeri cytochrome c (4) as a template. The protein shows unprecedented properties within the cytochrome c (4) family, including (1) an almost nonpolar surface charge distribution, (2) the absence of high-spin heme Fe(III) states, indicative of a thermodynamically stable and kinetically inert axial heme His,Met coordination, and (3) identical E degrees ' values for the two heme centers (+0.322 V vs the standard hydrogen elecrode). At pH extremes, both heme groups undergo the "acid" and "alkaline" conformational transitions typical of class I cytochromes c, involving ligand-exchange equilibria, whereas at intermediate pH values their electronic properties are sensitive to several residue ionizations. PMID- 18386081 TI - Structural basis for VO(2+)-inhibition of nitrogenase activity: (B) pH-sensitive inner-sphere rearrangements in the 1H-environment of the metal coordination site of the nitrogenase Fe-protein identified by ENDOR spectroscopy. AB - The nitrogenase Fe-protein is the specific ATP-activated electron donor to the active site-containing nitrogenase MoFe-protein. It has been previously demonstrated that different VO(2+)-nucleotide coordination environments exist for the Fe-protein that depend on pH and are distinguishable by EPR spectroscopy. After having studied the nitrogenase 31P and 23Na superhyperfine structure for this system by electron nuclear double resonance (ENDOR) spectroscopy (Petersen et al. 2008 in J Biol Inorg Chem. doi:10.1007/s00775-008-0360-0), we here report on the 1H-interactions with the nucleotide-bound metal center after substitution of the natural diamagnetic metal Mg2+ with paramagnetic oxo-vanadium(IV). ENDOR spectra show a number of resonances arising from interactions of the VO2+ ion with protons. In the presence of reduced Fe-protein and VO2+ ADP, at least three sets of nonexchangeable protons are detected. At low pH the superhyperfine couplings of most of these are consistent with proton interactions originating from the nucleotide. There is no indication of 1H-resonances that exchange in D2O at neutral pH and could be assigned to inner-sphere hydroxyl coordination. Exchangeable hydroxyl protons in the inner coordination sphere with reduced Fe protein are only found in the low pH form; based on their hyperfine tensor components these have been assigned to an axially coordinated hydroxyl water molecule. The pH-dependent alterations of the proton couplings that exchange in D2O suggest that they are partially caused by a rearrangement in the local hydroxyl coordination environment of the metal center. These rearrangements especially affect the apical metal position, where an axially coordinated water present at low pH is absent at neutral pH. Oxidation of the Fe-protein induced substantial changes in the electron-nucleus interactions. This indicates that the oxidation state of the iron-sulfur cluster has an important effect on the metal coordination environment at the nucleotide binding site of the Fe-protein. The distinct VO(2+)-nucleotide coordination structures with ADP and ATP and the redox state of the [4Fe-4S] cluster imply that VO2+ has a critical influence on the switch regions of the regulatory protein, and, taken together, this provides a plausible explanation for the inhibitory action of VO2+. PMID- 18386082 TI - Construction of three-dimensional tooth model by micro-computed tomography and application for data sharing. AB - The study of dental morphology is essential in terms of phylogeny. Advances in three-dimensional (3D) measurement devices have enabled us to make 3D images of teeth without destruction of samples. However, raw fundamental data on tooth shape requires complex equipment and techniques. An online database of 3D teeth models is therefore indispensable. We aimed to explore the basic methodology for constructing 3D teeth models, with application for data sharing. Geometric information on the human permanent upper left incisor was obtained using micro computed tomography (micro-CT). Enamel, dentine, and pulp were segmented by thresholding of different gray-scale intensities. Segmented data were separately exported in STereo-Lithography Interface Format (STL). STL data were converted to Wavefront OBJ (OBJect), as many 3D computer graphics programs support the Wavefront OBJ format. Data were also applied to Quick Time Virtual Reality (QTVR) format, which allows the image to be viewed from any direction. In addition to Wavefront OBJ and QTVR data, the original CT series were provided as 16-bit Tag Image File Format (TIFF) images on the website. In conclusion, 3D teeth models were constructed in general-purpose data formats, using micro-CT and commercially available programs. Teeth models that can be used widely would benefit all those who study dental morphology. PMID- 18386083 TI - Elongation cutoff technique: low-order scaling SCF method. AB - The elongation cutoff technique at restricted Hartree-Fock (HF) level of theory in conventional type of calculations, i.e., with two electron integrals (TEI) stored on a disc, is presented for two model systems. It is demonstrated that the number of TEI in the elongation cutoff calculations increases linearly with the system size thus, allowing to extend the conventional type of calculations to bigger systems. The step CPU (central processing unit) time in the elongation cutoff calculations is much lower than in the HF reference calculations. Such behavior reduces significantly the prefactor in the quadratic scaling relation. The total CPU time in the elongation calculation is about 40% lower than in the conventional HF calculations or comparable to direct type of calculations with the quantum fast multipoles method employed. It is shown that by introducing the interaction radius one can obtain linear scaling in the SCF calculations. Figure: The structure of density matrix and total CPU timings for polyglycine clusters in the elongation cutoff calculations. PMID- 18386084 TI - Catheter-related bacteremia from femoral and central internal jugular venous access. AB - The objective of this prospective observational study was to determine the influence of femoral and central internal jugular venous catheters on the incidence of catheter-related bacteremia (CRB). We included patients admitted to a 12-bed polyvalent medico-surgical intensive care unit over 4 years who received one or more femoral or central internal jugular venous catheters. We diagnosed 16 cases of CRB in 208 femoral catheters and 22 in 515 central internal jugular venous catheters. We found a higher incidence of CRB with femoral (9.52 per 1,000 catheter days) than with central internal jugular venous access (4.83 per 1,000 catheter days; risk ratio = 1.93; 95% confidence interval: 1.03-3.73; P = 0.04). Central internal jugular venous access could be considered a safer route of venous access than femoral access in minimizing the risk of central venous catheter-related bacteremia. PMID- 18386085 TI - Experimental study of meropenem in the therapy of cephalosporin-susceptible and resistant pneumococcal meningitis. AB - Meropenem is a carbapenem antibiotic that is highly active against the pathogens causing meningitis. Results with meropenem in the experimental rabbit model of penumococcal meningitis have been controversial, and the possible role of renal dehydropeptidase I in meropenem efficacy has been suggested. The aim of this study was to determine the efficacy of meropenem in two meningitis models and the possible influence of the animal model over results. Two strains of Streptococcus pneumoniae with different susceptibility to beta-lactams have been used in a guinea pig model and the classical rabbit meningitis model. Meropenem was bactericidal at 6 h in the guinea pig model against both strains with a reduction of >4 log ufc/ml. In the rabbit model it was bactericidal at 6 h against the susceptible strain, but against the resistant 3/8 therapeutical failures were recorded at 6 h, being bactericidal at 24 h. In conclusion, meropenem has shown bactericidal activity in both experimental models. This work emphasises the importance of an adequate election of the animal model for the appropriate development of studies of antimicrobial efficacy. We believe that guinea pig should be considered the best choice among laboratory animal species when assessing meropenem efficacy. PMID- 18386086 TI - State of the art trends in imaging renal of colic. AB - Radiology has a primary role in the work-up of renal colic, both to confirm urolithiasis and to help determine management. The traditional imaging has been conventional radiography and intravenous program with multidetector non-contrast enhanced helical computed tomography (NCECT) now the modality of choice. Nuclear medicine studies for renal colic are done now infrequently at most institutions. Ultrasound (US) is often done, especially in the emergency department, and magnetic resonance imaging (MRI) is promising. Radiation dose reduction is now on everyone's minds: Lower-dose CT techniques are being tested and used, and US and MR are considered as first modalities of choice in pregnant women and children. PMID- 18386087 TI - Automatic coil selection for channel reduction in SENSE-based parallel imaging. AB - OBJECTIVE: Coil arrays with large number of receive elements allow improved imaging performance and higher signal-to-noise-ratio. The MR systems supporting these arrays have to handle an increased amount of data and higher reconstruction burden. To overcome these problems, data reduction techniques need to be applied, realized either by linear combination of the original coil data prior to reconstruction or by discarding particular data from unimportant coil elements. MATERIALS AND METHODS: This work focuses on the latter approach and presents an efficient algorithm for automatic coil selection applicable to SENSE imaging. A singular value decomposition (SVD)-based coil selection is proposed that performs a coil element ranking quantifying the contribution of each coil element to the image reconstruction allowing appropriate coil selection. This approach makes use of the coil sensitivity information and takes reduction factor and phase encoding direction into account. RESULTS: Simulations, phantom and in vivo experiments were performed to validate the SVD-based coil selection algorithm. The proposed approach proved to be computationally efficient without remarkable image quality degradation. CONCLUSION: The SVD-based approach offers the opportunity for fast automatic coil selection. This could simplify clinical workflow and may, furthermore, pave the way for various 2D real-time and interventional applications. PMID- 18386088 TI - The characteristics of multifocal electroretinogram in isolated perfused porcine eye: cellular contributions to the in vitro porcine mfERG. AB - We investigated characteristics of multifocal electroretinograms (mfERG) from in vitro perfused porcine eyes. TTX, NMDA, APB, and PDA were used to identify contributions to the mfERG from inner retinal neurons, ON-pathway, OFF-pathway, and photoreceptors. The cellular contributions of the first-order kernel (K1) in an isolated perfused porcine mfERG came from both inner and outer retina, and were similar to those of in vivo porcine mfERG. In addition, application of isoflurane and propofol caused interference with the mfERG response which resembled inner retinal activities sensitive to TTX + NMDA application. Improved understanding of the cellular origins of the perfused porcine mfERG, in the absence of anesthetic agents, is useful for identifying changes shown in the waveform under anesthesia. PMID- 18386089 TI - Distribution of shear stress over smooth muscle cells in deformable arterial wall. AB - A biphasic, anisotropic model of the deformable aortic wall in combination with computational fluid dynamics is used to investigate the variation of shear stress over smooth muscle cells (SMCs) with transmural pressure. The media layer is modeled as a porous medium consisting of SMCs and a homogeneous porous medium of interstitial fluid and elastin, collagen and proteoglycans fibers. Interstitial fluid enters the media through fenestral pores, which are distributed over the internal elastic lamina (IEL). The IEL is considered as an impermeable barrier to fluid flow except at fenestral pores. The thickness and the radius of aortic wall vary with transmural pressure ranging from 10 to 180 mm Hg. It is assumed that SMCs are cylinders with a circular cross section at 0 mm Hg. As the transmural pressure increases, SMCs elongate with simultaneous change of cross sectional shape into ellipse according to the strain field in the media. Results demonstrate that the variation of shear stress within the media layer is significantly dependent on the configuration and cross sectional shape of SMCs. In the staggered array of SMCs, the shear stress over the first SMC nearest to the IEL is about 2.2 times lower than that of the square array. The shear stress even over the second nearest SMC to the IEL is considerably higher (about 15%) in the staggered array. In addition to configuration and cross sectional shape of SMCs, the variation of structural properties of the media layer with pressure and the sensitivity of the local shear stress to the minimum distance between SMCs and the IEL (reducing with transmural pressure) between SMCs and the IEL are studied. At 180 mm Hg, the ratio of the local shear stress of the nearest SMC to that of the second nearest SMC is 4.8 in the square array, whereas it reduces to about 1.8 in the staggered array. The importance of the fluid shear stress is associated with its role in the biomolecular state of smooth muscle cells bearing the shear stress. PMID- 18386090 TI - Chaotic behavior of the coronary circulation. AB - The regulation of the coronary circulation is a complex paradigm in which many inputs that influence vasomotor tone have to be integrated to provide the coronary vasomotor adjustments to cardiac metabolism and to perfusion pressure. We hypothesized that the integration of many disparate signals that influence membrane potential of smooth muscle cells, calcium sensitivity of contractile filaments, receptor trafficking result in complex non-linear characteristics of coronary vasomotion. To test this hypothesis, we measured an index of vasomotion, flowmotion, the periodic fluctuations of flow that reflect dynamic changes in resistances in the microcirculation. Flowmotion was continuously measured in periods ranging from 15 to 40 min under baseline conditions, during antagonism of NO synthesis, and during combined purinergic and NOS antagonism in the beating heart of anesthetized open-chest dogs. Flowmotion was measured in arterioles ranging from 80 to 135 microm in diameter. The signals from the flowmotion measurements were used to derive quantitative indices of non-linear behavior: power spectra, chaotic attractors, correlation dimensions, and the sum of the Lyapunov exponents (Kolmogorov-Sinai entropy), which reflects the total chaos and unpredictability of flowmotion. Under basal conditions, the coronary circulation demonstrated chaotic non-linear behavior with a power spectra showing three principal frequencies in flowmotion. Blockade of nitric oxide synthase or antagonism of purinergic receptors did not affect the correlation dimensions, but significantly increased the Kolmogorov-Sinai entropy, altered the power spectra of flowmotion, and changed the nature of the chaotic attractor. These changes are consistent with the view that certain endogenous controls, nitric oxide and various purines (AMP, ADP, ATP, adenosine) make the coronary circulation more predictable, and that blockade of these controls makes the control of flow less predictable and more chaotic. PMID- 18386091 TI - Dietary perilla oil inhibits proinflammatory cytokine production in the bronchoalveolar lavage fluid of ovalbumin-challenged mice. AB - To evaluate the anti-inflammatory effects of different dietary oils on ovalbumin sensitized and -challenged mice. Experimental BALB/c mice were fed with different diets containing 5% corn oil [rich in linoleic acid, 18:2n-6 polyunsaturated fatty acids (PUFA), as a control diet], 5% perilla oil (rich in alpha-linolenic acid, 18:3n-3 PUFA) or 5% compound oil containing 50% corn oil and 50% perilla oil, for 5 consecutive weeks. The leukocyte count, inflammatory mediators, and cytokine levels, including proinflammatory and Th1/Th2 cytokines in the bronchoalveolar lavage fluid (BALF) from the mice were determined. The results showed that 5% compound oil administration significantly (P < 0.05) decreased eosinophilic infiltration. Dietary perilla oil could not significantly (P > 0.05) decrease the eosinophil accumulation or the secretions of inflammatory mediators such as prostaglandin E2 (PGE2), histamine, nitric oxide and eotaxin. However, dietary perilla oil significantly (P < 0.05) reduced proinflammatory cytokine (TNF-alpha, IL-1beta and IL-6) and Th1 cytokine (IFN-gamma and IL-2) production. The production of Th2 cytokine IL-10, but not IL-4 and IL-5, was also significantly inhibited by perilla oil administration. The results suggest that dietary perilla oil might alleviate inflammation via decreasing the secretion of pro-inflammatory cytokines in BALF, but failed to regulate the Th1/Th2 balance toward Th1 pole during the Th2-skewed allergic airway inflammation. PMID- 18386092 TI - Red light of 647 nm enhances osteogenic differentiation in mesenchymal stem cells. AB - The use of light for medical treatment has been studied previously. In this study, we examined the effect of light from a red light-emitting diode on osteogenic differentiation of mouse mesenchymal stem cells (D1 cells) which were cultured in the presence of osteogenic differentiation medium (ODM) for 3 days, then exposed to a red light-emitting diode (LED) light of 647 nm wavelength once for 10 s, 30 s or 90 s with radiation energies of 0.093 J, 0.279 J and 0.836 J, respectively. D1 cells in the presence of ODM differentiated into osteoblasts, and this process was enhanced on exposure to LED light in ODM medium. This effect was confirmed by increased Alizarin red staining, higher alkaline phosphatase (ALP) activity, higher mRNA expressions of osteocalcin, collagen type I, osteopontin and Runt-related transcription factor2 (Runx2), and higher levels by reverse transcriptase-polymerase chain reaction (RT-PCR) and by increased immunofluorescence staining against cluster of differentiation 44 (CD44) by immunofluorescence microscopy, confocal microscopy and flow cytometric analysis. These data suggest that osteogenic differentiation of mesenchymal stem cells (MSCs) in ODM is enhanced by LED light exposure. PMID- 18386093 TI - Evaluation of collagen in atherosclerotic plaques: the use of two coherent laser based imaging methods. AB - Acute coronary events such as myocardial infarction are frequently caused by the rupture of unstable atherosclerotic plaque. Collagen plays a key role in determining plaque stability. Methods to measure plaque collagen content are invaluable in detecting unstable atherosclerotic plaques. Recently, novel coherent laser-based imaging techniques, such as polarization-sensitive optical coherence tomography (PSOCT) and laser speckle imaging (LSI) have been investigated, and they provide a wealth of information related to collagen content and plaque stability. Additionally, given their potential for intravascular use, these technologies will be invaluable for improving our understanding of the natural history of plaque development and rupture and, hence, enable the detection of unstable plaques. In this article we review recent developments in these techniques and potential challenges in translating these methods into intra-arterial use in patients. PMID- 18386094 TI - Computer-aided image analysis of focal hepatic lesions in ultrasonography: preliminary results. AB - PURPOSE: To develop a computer-aided image analysis (CAIA) algorithm for analyzing US features of focal hepatic lesions and to correlate the feature values of CAIA with radiologists' grading. MATERIALS AND METHODS: Two abdominal radiologists, blinded to the final diagnosis, independently evaluated sonographic images of 51 focal hepatic lesions in 47 patients: hemangiomas (n = 19), hepatic simple cysts or cystic lesions (n = 14), hepatocellular carcinoma (n = 11), metastases (n = 6), and focal fat deposition (n = 1). All images were graded using a 3- to 5-point scale, in terms of border (roundness, sharpness, and the presence of peripheral rim), texture (echogenicity, homogeneity, and internal artifact), posterior enhancement, and lesion conspicuity. Using a CAIA, texture and morphological parameters representing radiologists' subjective evaluations were extracted. Correlations between the radiologists and the CAIA for assessing parameters in corresponding categories were computed by means of weighted kappa statistics and Spearman correlation test. RESULTS: A good agreement was achieved between CAIA and radiologists for grading echogenicity (weighted kappa = 0.675) and the presence of hyper- or hypoechoic rim (weighted kappa = 0.743). Several CAIA-derived features representing homogeneity of the lesions showed good correlations (correlation coefficient (gamma) = 0.603 approximately 0.641) with radiologists' grading (P < 0.05). For internal artifact (gamma = 0.469-0.490) and posterior enhancement (gamma = -0.516) of the cyst and lesion conspicuity (gamma = 0.410), a fair correlation between CAIA and radiologists was obtained (P < 0.05). However, parameters representing lesions' border such as sharpness (gamma = 0.252-0.299) and roundness (gamma = -0.134-0.163) showed no significant correlation (P > 0.05). CONCLUSION: As a preliminary step in US computer-aided diagnosis for focal hepatic lesions, a CAIA algorithm was constructed with a good agreement and correlation with human observers in most US features. In addition, these features should be weighted highly when a computer-aided diagnosis for characterizing focal liver lesions on US is designed and developed. PMID- 18386096 TI - Increased terpenoid accumulation in cotton (Gossypium hirsutum) foliage is a general wound response. AB - The subepidermal pigment glands of cotton accumulate a variety of terpenoid products, including monoterpenes, sesquiterpenes, and terpenoid aldehydes that can act as feeding deterrents against a number of insect herbivore species. We compared the effect of herbivory by Spodoptera littoralis caterpillars, mechanical damage by a fabric pattern wheel, and the application of jasmonic acid on levels of the major representatives of the three structural classes of terpenoids in the leaf foliage of 4-week-old Gossypium hirsutum plants. Terpenoid levels increased successively from control to mechanical damage, herbivory, and jasmonic acid treatments, with E-beta-ocimene and heliocide H1 and H4 showing the highest increases, up to 15-fold. Herbivory or mechanical damage to older leaves led to terpenoid increases in younger leaves. Leaf-by-leaf analysis of terpenes and gland density revealed that higher levels of terpenoids were achieved by two mechanisms: (1) increased filling of existing glands with terpenoids and (2) the production of additional glands, which were found to be dependent on damage intensity. As the relative response of individual terpenoids did not differ substantially among herbivore, mechanical damage, and jasmonic acid treatments, the induction of terpenoids in cotton foliage appears to represent a non-specific wound response mediated by jasmonic acid. PMID- 18386097 TI - (E,E)-alpha-farnesene, an alarm pheromone of the termite Prorhinotermes canalifrons. AB - The behavioral and electroantennographic responses of Prorhinotermes canalifrons to its soldier frontal gland secretion, and two separated major components of the secretion, (E)-1-nitropentadec-1-ene and (E,E)-alpha-farnesene, were studied in laboratory experiments. Behavioral experiments showed that both the frontal gland secretion and (E,E)-alpha-farnesene triggered alarm reactions in P. canalifrons, whereas (E)-1-nitropentadec-1-ene did not affect the behavior of termite groups. The alarm reactions were characterized by rapid walking of activated termites and efforts to alert and activate other members of the group. Behavioral responses to alarm pheromone differed between homogeneous and mixed groups, suggesting complex interactions. Antennae of both soldiers and pseudergates were sensitive to the frontal gland secretion and to (E,E)-alpha-farnesene, but soldiers showed stronger responses. The dose responses to (E,E)-alpha-farnesene were identical for both soldiers and pseudergates, suggesting that both castes use similar receptors to perceive (E,E)-alpha-farnesene. Our data confirm (E,E)-alpha farnesene as an alarm pheromone of P. canalifrons. PMID- 18386098 TI - Effects of p-vinylphenyl glycosides and other related compounds on the oviposition behavior of Ceratitis capitata. AB - Elaphoside-A [p-vinylphenyl (beta-D: -glucopyranosyl)-(1-->3)-beta-D: allopyranoside], a Mediterranean fruit fly oviposition deterrent, was previously isolated from an Argentine collection of the fern Elaphoglossum piloselloides. In order to establish the structural requirements for the observed oviposition inhibition, we synthesized and characterized 4 known and 21 new aromatic glycosides structurally related to elaphoside-A. Their effects on the oviposition behavior of Ceratitis capitata females are discussed. PMID- 18386099 TI - Analytical results for individual and group selection of any intensity. AB - The idea of evolutionary game theory is to relate the payoff of a game to reproductive success (= fitness). An underlying assumption in most models is that fitness is a linear function of the payoff. For stochastic evolutionary dynamics in finite populations, this leads to analytical results in the limit of weak selection, where the game has a small effect on overall fitness. But this linear function makes the analysis of strong selection difficult. Here, we show that analytical results can be obtained for any intensity of selection, if fitness is defined as an exponential function of payoff. This approach also works for group selection (= multi-level selection). We discuss the difference between our approach and that of inclusive fitness theory. PMID- 18386100 TI - A systematic review of teamwork training interventions in medical student and resident education. AB - BACKGROUND: Teamwork is important for improving care across transitions between providers and for increasing patient safety. OBJECTIVE: This review's objective was to assess the characteristics and efficacy of published curricula designed to teach teamwork to medical students and house staff. DESIGN: The authors searched MEDLINE, Education Resources Information Center, Excerpta Medica Database, PsychInfo, Cumulative Index of Nursing and Allied Health Literature, and Scopus for original data articles published in English between January 1980 and July 2006 that reported descriptions of teamwork training and evaluation results. MEASUREMENTS: Two reviewers independently abstracted information about curricular content (using Baker's framework of teamwork competencies), educational methods, evaluation design, outcomes measured, and results. RESULTS: Thirteen studies met inclusion criteria. All curricula employed active learning methods; the majority (77%) included multidisciplinary training. Ten curricula (77%) used an uncontrolled pre/post design and 3 (23%) used controlled pre/post designs. Only 3 curricula (23%) reported outcomes beyond end of program, and only 1 (8%) >6 weeks after program completion. One program evaluated a clinical outcome (patient satisfaction), which was unchanged after the intervention. The median effect size was 0.40 (interquartile range (IQR) 0.29, 0.61) for knowledge, 0.38 (IQR 0.32, 0.41) for attitudes, 0.41 (IQR 0.35, 0.49) for skills and behavior. The relationship between the number of teamwork principles taught and effect size achieved a Spearman's correlation of .74 (p = .01) for overall effect size and .64 (p = .03) for median skills/behaviors effect size. CONCLUSIONS: Reported curricula employ some sound educational principles and appear to be modestly effective in the short term. Curricula may be more effective when they address more teamwork principles. PMID- 18386095 TI - Rainbow smelt (Osmerus mordax) genomic library and EST resources. AB - Genomic resources in rainbow smelt (Osmerus mordax) enable us to examine the genome duplication process in salmonids and test hypotheses relating to the fate of duplicated genes. They further enable us to pursue physiological and ecological studies in smelt. A bacterial artificial chromosome library containing 52,410 clones with an average insert size of 146 kb was constructed. This library represents an 11-fold average coverage of the rainbow smelt (O. mordax) genome. In addition, several complementary deoxyribonucleic acid libraries were constructed, and 36,758 sequences were obtained and combined into 12,159 transcripts. Over half of these transcripts have been identified, several of which have been associated with cold adaptation. These basic resources show high levels of similarity (86%) to salmonid genes and provide initial support for genome duplication in the salmonid ancestor. They also facilitate identification of genes important to fish and direct us toward new technologies for other studies in fish biology. PMID- 18386101 TI - Effects of disclosing financial interests on attitudes toward clinical research. AB - BACKGROUND: The effects of disclosing financial interests to potential research participants are not well understood. OBJECTIVE: To examine the effects of financial interest disclosures on potential research participants' attitudes toward clinical research. DESIGN AND PARTICIPANTS: Computerized experiment conducted with 3,623 adults in the United States with either diabetes mellitus or asthma, grouped by lesser and greater severity. Respondents read a description of a hypothetical clinical trial relevant to their diagnosis that included a financial disclosure statement. Respondents received 1 of 5 disclosure statements. MEASUREMENTS: Willingness to participate in the hypothetical clinical trial, relative importance of information about the financial interest, change in trust after reading the disclosure statement, surprise regarding the financial interest, and perceived effect of the financial interest on the quality of the clinical trial. RESULTS: Willingness to participate in the hypothetical clinical trial did not differ substantially among the types of financial disclosures. Respondents viewed the disclosed information as less important than other factors in deciding to participate. Disclosures were associated with some respondents trusting the researchers less, although trust among some respondents increased. Most respondents were not surprised to learn of financial interests. Researchers owning equity were viewed as more troubling than researchers who were compensated for the costs of research through per capita payments. CONCLUSIONS: Aside from a researcher holding an equity interest, the disclosure to potential research participants of financial interests in research, as recommended in recent policies, is unlikely to affect willingness to participate in research. PMID- 18386102 TI - Cushing's syndrome due to ectopic adrenocorticotropic hormone production secondary to hepatic carcinoid: diagnosis, treatment, and improved quality of life. AB - INTRODUCTION: We describe a previously healthy 40-year-old woman with Cushing's syndrome caused by adrenocorticotropic hormone (ACTH) secretion from metastatic carcinoid. CASE REPORT: Over a 2-year period, the patient had multiple hospitalizations for uncontrolled hypertension, hyperglycemia, and hypokalemia. She had transient flushing, rashes, and a rapid weight gain. In addition, she developed anasarca and had a nontraumatic hip fracture 1 month before presentation. Subsequently, a hypertensive crisis resulted in admission to the intensive care unit and fine-needle aspiration of a liver lesion. DISCUSSION: A diagnosis of metastatic carcinoid was established. She was transferred to our hospital for further evaluation and management. On arrival, she had the signs of Cushing's syndrome. Despite extensive evaluation, her primary carcinoid tumor was not localized. She was treated successfully with bilateral adrenalectomy and octreotide. CONCLUSION: This case illustrates how early recognition of the signs and symptoms of excess ACTH is important for prompt and appropriate treatment. PMID- 18386103 TI - Physician reminders to promote surveillance colonoscopy for colorectal adenomas: a randomized controlled trial. AB - BACKGROUND: Most colorectal cancers develop from adenomatous polyps. National guidelines recommend surveillance colonoscopy within 5 years after such polyps are removed. OBJECTIVE: To determine whether surveillance colonoscopy can be increased among overdue patients by reminders to their primary physicians. DESIGN: Randomized, controlled trial of patient-specific reminders mailed to 141 physicians in 2 Massachusetts primary care networks during April, 2006. PATIENTS: Seven hundred seventeen patients who had colorectal adenomas removed during 1995 through 2000 and no follow-up colonoscopy identified via automated review of electronic records through March, 2006. MEASUREMENTS AND MAIN RESULTS: The use of colonoscopy and detection of new adenomas or cancer were assessed at 6 months by a blinded medical record review in all patients. Among 358 patients whose physicians received reminders, 33 (9.2%) patients underwent colonoscopy within 6 months, compared with 16 (4.5%) of 359 patients whose physicians did not receive reminders (P = 0.009). In prespecified subgroups, this effect did not differ statistically between 2 primary care networks, elderly and nonelderly patients, or women and men (all P > 0.60 by Breslow-Day test). New adenomas or cancer were detected in 14 (3.9%) intervention patients and 6 (1.7%) control patients (P = 0.06), representing 42.4% and 37.5% of patients who underwent colonoscopy in each group, respectively. Despite using advanced electronic health records to identify eligible patients, 22.5% of enrolled patients had a prior follow-up colonoscopy ascertained only by visual record review, and physicians reported 27.9% of intervention patients were no longer active in their practice. CONCLUSIONS: Among patients with prior colorectal adenomas, physician reminders increased the use of surveillance colonoscopy, but better systems are needed to identify eligible patients (ClinicalTrials.gov ID number NCT00397969). PMID- 18386104 TI - A prospective evaluation of a variant of biliopancreatic diversion with Roux-en-Y reconstruction in mega-obese patients (BMI > or = 70 kg/m(2)). AB - BACKGROUND: Super-obese patients with BMI > or = 70 kg/m(2) present a special subgroup among the obese population due to the higher incidence of severe comorbidities and increased technical difficulties in perioperative management. The optimal surgical approach still remains controversial. METHODS: From January 2002 to October 2007, 68 super-obese patients with BMI > or = 70 kg/m(2) (75.7 +/ 5.61) and various comorbidities (mean 1.45 +/- 1, range 0-5), underwent a variant of biliopancreatic diversion with Roux-en-Y reconstruction (BPD-RYGBP) at our institution. The mean age was 39.6 +/- 9.5 years, and the excess weight was 146.1 +/- 24.5 kg. The safety and the effectiveness of the procedure were prospectively evaluated. RESULTS: Thirty-one patients have completed their second postoperative year. Mean BMI after 2 years follow-up was 35 +/- 5.33 kg/m(2) (p < 0.001) and mean %EWL was 79.9% (range 57.17-149.89). Significant improvement in preexisting comorbidities was also observed (mean 0.95 +/- 0.1, range 0-2; p < 0.001). Early major postoperative complications included one case of acute renal failure (1.47%), three anastomotic leaks (4.41%), four pulmonary embolisms (5.88%), one hemorrhage requiring reoperation (1.47%), and three cases of pneumonia (4.41%), resulting in a total morbidity rate of 8.82% and a mortality rate of 8.82%. Late complications included small bowel obstruction in four patients (5.88%), pulmonary embolism in one patient (1.47%), severe hypoalbuminemia requiring artificial nutrition support in six patients (8.82%), and incisional hernia in 23 patients (33.8%). Late morbidity was 41.1%, and there was no late mortality. CONCLUSION: BPD-RYGBP appears to be an effective procedure in patients with BMI > or = 70 kg/m(2) providing adequate weight loss and improvement of co-existing comorbidies after 2 years. Nevertheless, it is associated with higher morbidity and mortality rates compared to patients with BMI < 70 kg/m(2) undergoing the same surgical procedure. PMID- 18386105 TI - An individualized approach to abdominoplasty in the presence of bilateral subcostal scars after open gastric bypass. AB - BACKGROUND: Patients requiring surgical skin excision after massive weight loss are challenging and require an individualized approach. The characteristic abdominal deformity includes a draping apron of panniculus, occasionally associated with previous transverse surgical scars from open gastric bypass surgery in the upper abdomen, which compromise blood supply of the abdominal skin. METHODS: We propose four different surgical techniques for safe abdominal body contouring in the presence of such scars: (1) a limited abdominoplasty of the lower abdomen is performed, and then contouring is completed by a reversed abdominoplasty with scar positioning in the submammary folds; (2) a one-stage procedure characterized by skin resection in the upper and lower abdomen, in which blood supply of the skin island between the submammary and suprapubic incisions is ensured by periumbilical perforators; (3) a perforator-sparing abdominoplasty with selective dissection of periumbilical abdominal wall perforators to secure flap blood supply and allow complete flap undermining up to the xyphoid process; (4) for patients with extensive excess skin, a modified Fleur-de-Lys abdominoplasty performed in such a way that the old transverse scar is transformed into a vertical scar. RESULTS: The treatment of four exemplary patients is described. All techniques yielded good esthetic and functional results through preservation of abdominal blood supply. CONCLUSION: Through an individualized approach, adequate abdominal body contouring can be performed safely, even in the presence of transverse surgical scars in the upper abdomen. PMID- 18386106 TI - Is preoperative manometry in restrictive bariatric procedures necessary? AB - BACKGROUND: Restrictive bariatric procedures are frequently considered for patients with morbid obesity, because the weight loss and reduction of comorbidities are good. An impact on gastroesophageal reflux disease (GERD), which is common in this population, may be anticipated. Converse results of GERD symptoms are reported for patients after adjustable gastric banding (AGB), sleeve gastrectomy (SG), and Roux-en-Y gastric bypass (RYGBP). METHODS: A literature search was performed and, with our personal experience, are summarized. RESULTS: Esophageal manometry is a practical tool to identify functional disorders of the esophageal body and the lower esophageal sphincter (LES). For patients with weak esophageal body motility, AGB should not be considered as a therapeutic option because esophageal dilation, esophageal stasis, and consequent esophagitis often occur during long-term follow-up, and band deflation is inevitable. Stable body weight can therefore not be achieved in these patients. Low resting pressure of the LES may be a contraindication for SG, because taking away the angle of His further impairs the antireflux mechanism at the cardia. So far, RYGBP is an option for all morbidly obese patients regardless of the results of esophageal manometry. CONCLUSION: Preoperative esophageal manometry is advised for restrictive procedures such as AGB and SG. PMID- 18386107 TI - The relationship between body mass index and postoperative mortality from critical illness. AB - BACKGROUND: Conflicting data exist regarding the effect of body mass index (BMI) on postoperative mortality from critical illness. Few studies have examined this issue in surgical patients specifically. We tested the hypothesis that BMI is associated with mortality from surgical critical illness. METHODS: Consecutive admissions to a university surgical intensive care unit (SICU) were analyzed from January 2005-August 2006. Admission BMI was analyzed as both a five-level categorical (underweight, <18.5 kg/m2; normal weight, 18.5-24.9 kg/m2; overweight, 25.0-29.9 kg/m2; obese, 30.0-39.9 kg/m2; morbidly obese, > or =40 kg/m2) and dichotomous (> or =40 kg/m2 vs. <40 kg/m2) variable among all patients as well as a subgroup of patients with a SICU length of stay (ULOS) > or =4 days. Multivariable logistic regression models were fit to determine the independent effect of BMI group on SICU mortality. RESULTS: The total sample size was 946, with 490 patients admitted to the SICU for > or =4 days (51.8%). Of the variables tested, age, acute physiology and chronic health evaluation III score, gender, diabetes mellitus, and need for insulin infusion varied significantly among the five BMI groups. After adjustment for these variables, BMI was not predictive of mortality when analyzed as either a five-level categorical or dichotomous variable, regardless of ULOS. CONCLUSION: BMI is not related to mortality of surgical critical illness. Several factors, including modern ICU care, may mitigate the risks of obesity in the SICU. PMID- 18386108 TI - Post discharge prophylactic anticoagulation in gastric bypass patient-how safe? AB - BACKGROUND: It is becoming an increasingly common practice to discharge gastric bypass (GBP) patients on prophylactic anticoagulation. This is because pulmonary embolism (PE) is a common cause of mortality postoperatively. This study was undertaken to: (1) determine the incidence of major bleeding in GBP patients discharged on prophylactic low molecular weight heparin (LMWH)-enoxaparin and, (2) correlate the bleeding risk to the dose used. METHODS: Retrospective chart review of all open GBP operation from June 2004 to August 2005. One hundred and twenty seven patients were sent home on LMWH for 2 weeks. INDICATIONS: Body mass index (BMI) > or =50 kg/m(2) with chronic venous stasis and/or obstructive sleep apnea, previous history of PE or deep vein thrombosis (DVT) or BMI > or =60 kg/m(2). The study group was divided into two subgroups: 40 mg twice daily (bid) and 60 mg bid LMWH. Statistical analysis was done with the chi-square. The primary outcome measure was major bleeding; defined as bleeding during the period of LMWH use associated with symptomatic decrease in hematocrit (HCT), necessitating stopping LMWH administration before the end of the study period (2 weeks), bleeding-related readmission, blood transfusion, or intervention. Excluded were patients on warfarin or treated with therapeutic LMWH. RESULTS: The groups were similarly matched for age, body mass index, and risk factors. No episode of major bleeding after discharge occurred in either group. CONCLUSION: The use of low molecular weight heparin for prophylactic anticoagulation after open gastric bypass is not associated with risk of major bleeding. PMID- 18386109 TI - Management of obesity in the writings of Soranus of Ephesus and Caelius Aurelianus. AB - Caelius Aurelianus (5th century AD) is famous for translating the writings of Soranus of Ephesus (2nd century AD) into Latin. In his book On acute and chronic diseases, Caelius gives an almost complete edition of the works of Soranus, while adding some of his own and other writers' material. Obesity was even at that time considered to be a disease and the authors treat the subject extensively. The nature, etiology, the clinical manifestations of obesity are described, although treatment covers the greatest part of the material. Infliction of perspiration, very low caloric intake diet, exercise, and diuresis are proposed, and one cannot overlook the strong similarities of some of the treatments with those used nowadays as secondary aids for weight loss. PMID- 18386110 TI - Bariatric surgery: low mortality at a high-volume center. AB - BACKGROUND: The American Society of Bariatric Surgery has initiated a Bariatric Surgery Center of Excellence Program and the American College of Surgeons has followed with their Bariatric Surgery Center Network Accreditation Program. These programs postulate that concentration of weight loss operations in high-volume centers will decrease surgical mortality and improve outcomes. METHODS: The purpose of this study was to calculate the in-hospital mortality for bariatric operations accomplished at the highest volume bariatric surgery center in the state of New Jersey. After receiving Institutional Revew Board approval, the revised surgical schedule was used to identify all patients undergoing weight loss surgery (WLS) at Hackensack University Medical Center from 1998 through June, 2006. Data for these patients were then harvested from the hospital's electronic medical record. Step-wise and univariate logistic regression analysis tested the impact of various factors on hospital length of stay and in-hospital mortality. RESULTS: Between 1998 and June, 2006, 5,365 patients underwent WLS surgery: 2,099 open vertical banded gastroplasty-Roux en Y gastric bypass (VBG RYGB); 2,177 laparoscopic Roux en Y gastric bypass (LRYGB); and 1,089 laparoscopic adjustable gastric banding (LAGB). 75.5% of patients were women. Median age was 41 years old (13-79), median weight 128 kg (81.2-290.3), and median body mass index 46.1 kg/m2 (35.0-92.6). Median total operating room time for VBG-RYGB was 115 min (33-328); LRYGB 155 min (53-493), and LAGB 92 min (33 274). Median length of stay for VBG-RYGB was 3 days (1-39 days), LRYGB 2 days (1 46 days), and LAGB 1 day (1-20). Seven patients died in hospital after the 5,365 WLS operations (0.13%): four after VBG-RYGB (0.19%); three after LRYGB (0.14%); and none after LAGB (0%). The characteristics of the patients who died did not significantly differ from the group as a whole. CONCLUSION: Surgeons at Hackensack University Medical Center, a high volume, accredited 1A American College of Surgeons Bariatric Surgery Center, achieved a 0.13% mortality among 5,365 patients undergoing weight loss operations between 1998 and June, 2006. This study supports the concept that high-volume centers perform bariatric operations with low mortalities. PMID- 18386111 TI - Monitored anesthesia care with propofol versus surgeon-monitored sedation with benzodiazepines and narcotics for preoperative endoscopy in the morbidly obese. AB - BACKGROUND: Although still controversial, upper endoscopy is frequently performed before bariatric surgery. This study investigated the hypothesis that morbidly obese patients would prefer anesthesiologist-monitored sedation (AMS) compared to surgeon-monitored sedation (SMS) during preoperative endoscopy. METHODS: All patients who underwent endoscopy before their bariatric surgery were given a post procedure survey regarding their experience with the preoperative endoscopy. The survey inquired about issues during and after the procedure. We compared patients who had AMS with IV propofol versus SMS IV narcotics and benzodiazepines. RESULTS: There were 100 patients (SMS=49 and AMS=51). Few patients complained of pain in the abdomen or throat during the procedure (AMS vs. SMS=2 vs. 8% and 2 vs. 10%, respectively; p=NS). More patients complained about throat pain after the procedure (AMS vs. SMS=37 vs. 45%; p=NS). More patients in the SMS group remembered the scope being placed in the mouth versus AMS (33 vs. 10%; p<0.02). More patients remembered gagging during the procedure in the SMS group versus the AMS group, but this did not reach statistical significance (24 vs. 10%; p=0.06). There was a trend that more patients in the AMS group felt they recovered in less than 1 h (53%) compared to the SMS group (37%; p=0.1). CONCLUSION: Patients who undergo upper endoscopy with either AMS or SMS seem to tolerate the procedure well. The preliminary benefits seen with AMS need to be further explored. AMS should be considered for patients undergoing preoperative upper endoscopy before bariatric surgery. PMID- 18386112 TI - A new technique to reattach an extended trochanteric osteotomy in revision THA using suture cord. AB - The extended trochanteric osteotomy (ETO) is widely used for revision total hip arthroplasty (THA). We describe a novel technique to reattach the extended trochanteric osteotomy using a suture cord. We retrospectively reviewed 20 patients who underwent revision by this technique with a minimum followup of 2 years (mean, 2.2 years; range, 2-4 years). Bony union of the osteotomy occurred in 19 patients and fibrous union in one patient who remained asymptomatic. We observed proximal migration of the osteotomy fragment in one patient (5 mm) who had bony union of the osteotomy. This technique of anchoring the osteotomy fragment directly onto the prosthetic stem using a suture cord provides a secure fixation and does not endanger the blood supply of the osteotomy. It also has the advantage of avoiding the morbidity associated with cerclage wires and the costs associated with cabling systems. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 18386113 TI - Physical sequelae and depressive symptoms in gynecologic cancer survivors: meaning in life as a mediator. AB - BACKGROUND: Continuing symptoms and poor health following cancer treatments may alter meaning in life for cancer survivors. Gynecologic cancer survivors are particularly troubled with physical sequelae. In addition, for the most common sites of disease, such as breast and gynecologic cancers, the prevalence of depression is also high. PURPOSE: This study tests meaning in life as a mechanism for the relationship between physical symptoms and depressive symptoms. METHODS: Gynecologic cancer survivors (N = 260) participated. Measures of physical sequelae (nurse rated symptoms/signs, patient-reported gynecologic symptoms), meaning in life (harmony, life purpose, spirituality, and conversely, confusion and loss), and depressive symptoms were obtained at the time of a routine clinical follow-up visit 2-10 years following the completion of treatment. Latent variables were defined, and structural equation modeling tested a mediator model. RESULTS: Analyses support partial mediation. That is, survivors with more physical sequelae also reported lower levels of meaning in life, which was associated with higher levels of depressive symptoms. CONCLUSIONS: Gynecologic cancer patients have been neglected in psychosocial research, and findings highlight the importance of existential issues in their lives. While many adjust well, those with persistent physical functioning deficits may experience depressive symptoms. By appreciating the role of meaning in their experience, we may help survivors foster their own growth and perspectives important for their future. PMID- 18386114 TI - Tuberin, p27 and mTOR in different cells. AB - Mutations in the genes TSC1 or TSC2 cause the autosomal dominantly inherited tumor suppressor syndrome tuberous sclerosis, which is characterized by the development of tumors, named hamartomas, in different organs. The TSC gene products, hamartin and tuberin, form a complex, of which tuberin is assumed to be the functional component. Both, hamartin and tuberin have been implicated in the control of the cell cycle by activating the cyclin-dependent kinase inhibitor p27 and in cell size regulation by inhibiting the mammalian target of rapamycin (mTOR) a regulator of the p70 ribosomal protein S6 kinase (p70S6K) and its target the ribosomal protein S6. The tuberin/hamartin complex was shown to protect p27 from protein degradation. Within the mTOR signaling pathway tuberin harbors GTPase activating (GAP) potential toward Rheb, which is a potent regulator of mTOR. In this study, we have analyzed the protein levels of tuberin, p27, cyclin D1, mTOR and phospho mTOR Ser2448 (activated mTOR), S6 and phospho S6 Ser240/244 (activated S6) and as controls alpha-tubulin and topoisomerase IIbeta, in ten different cells, including primary normal cells, immortalized and transformed cell lines. PMID- 18386115 TI - Screening of BCS1L mutations in severe neonatal disorders suspicious for mitochondrial cause. AB - The BCS1L gene encodes a chaperone responsible for assembly of respiratory chain complex III (CIII). A homozygous point mutation (232A-->G) has been found as the genetic etiology for fetal growth retardation, amino aciduria, cholestasis, iron overload, lactic acidosis, and early death (GRACILE) syndrome (MIM 603358). Variable phenotypes have been found with other mutations. Our aim was to assess whether 232A-->G or other BCS1L mutations were present in infants (n = 21) of Finnish origin with severe, lethal disease compatible with mitochondrial disorder. A further aim was to confirm the GRACILE genotype-phenotype constancy (n = 8). Three new cases with homozygous 232A-->G mutation were identified; all had the primary GRACILE characteristics. No other mutations were found in the gene in other cases. All infants with GRACILE syndrome had the typical mutation. In conclusion, the rather homogenous population of Finns seems to have a specific BCS1L mutation that, as homozygous state, causes GRACILE syndrome, whereas other mutations are rare or not occurring. Thus, the novel clinical implication of this study is to screen for BCS1L mutations only if CIII is dysfunctioning or lacking Rieske protein, and to assess 232A-->G mutation in cases with GRACILE syndrome. PMID- 18386116 TI - A peritoneal-based automated wearable artificial kidney. AB - Work on wearable kidneys has evolved around the technology of hemodialysis or hemofiltration, which call for continuous anticoagulation of the extracoporeal circulation and are encumbered with potential immunologic and non-immunologic complications of continuous blood-artificial membrane interactions. A peritoneal based automated wearable artificial kidney (AWAK) requires no extracorporeal circulation and is therefore "bloodless." Because AWAK is designed to continuously regenerate and reuse the spent dialysate in perpetuity, it is also "waterless." A sorbent-based assembly regenerates both the aqueous and the protein components (AqC and PrC) of the spent dialysate, producing a novel, autologous protein-containing dialysate. The regenerated AqC has the same composition as the commercially available peritoneal dialysate, but contains bicarbonate instead of lactate and has a more physiological pH. The regenerated PrC is recycled back into the peritoneal cavity, thereby ameliorating or eliminating protein loss. Depending on the steady-state protein concentrations that can be achieved (under the condition of continuous dialysate regeneration and recycling), the PrC also has the potential of both augmenting ultrafiltration and mediating the removal of protein-bound toxins. Additional sorbents can be incorporated into AWAK for the removal of middle molecular weight uremic toxins. At a regeneration rate of 4 l/h, AWAK provides a dialysate flow of 96 l/day (8-12 times the current rate). Round-the-clock dialysis and ultrafiltration provide steady-state metabolic-biochemical and fluid balance regulation, thereby eliminating "shocks" of abrupt changes in these parameters that characterize the current dialytic modalities. Dialysis-on-the-go, made possible by AWAK's "wearability" and automation, frees end-stage renal failure patients from the servitude that is demanded by the current dialytic regimentations. PMID- 18386117 TI - Monkeys with disabilities: prevalence and severity of congenital limb malformations in Macaca fuscata on Awaji Island. AB - The Awajishima Monkey Center (AMC) free-ranging, provisioned population of Japanese macaques has included individuals with congenital limb malformations (CLMs) for at least 40 years. Including new data from this study, 16.1% of AMC infants from 1969 to 2007 (185 of 1,150) were born with CLMs. However, relatively little is known about the demographics of CLMs in the population, particularly the relationships among occurrence and severity of CLMs and age-sex demographics after infancy. In 2004, we conducted a census at AMC. Of the 199 monkeys censused, 34 individuals (17.1%) had CLMs. To estimate the severity of CLMs, we created an index that ranks individuals on a scale of 0 to 1 based on affected and absent limbs and digits. The severity of CLMs varied greatly (index range = 0.01-0.79, mean = 0.29), with similar variation in severity in each age-sex class (Student t-test, P > 0.05). PMID- 18386118 TI - A case of recurrent invasive lobular carcinoma of the breast found as metastasis to the duodenum. AB - A 57-year-old woman underwent modified radical mastectomy for cancer of the left breast (stage IIB) in February 2004. Invasive lobular carcinoma was diagnosed on histopathological examination. The patient received postoperative chemotherapy and endocrine therapy on an outpatient basis and was observed. In August 2005, anorexia developed. Blood chemical tests showed elevated levels of liver enzymes and bilirubin. Computed tomography (CT) of the abdomen revealed an enlarged duodenum and dilated intrahepatic biliary and pancreatic ducts. Upper gastrointestinal endoscopy showed edema of the duodenum. A biopsy yielded a diagnosis of poorly differentiated adenocarcinoma. Duodenal carcinoma was suspected, and a pancreatoduodenectomy was performed. Duodenal metastasis from invasive lobular carcinoma was diagnosed on postoperative histopathological examination. After surgery, the patient recovered uneventfully and was discharged from the hospital. In March 2006, bilateral hydronephrosis apparently caused by peritoneal metastasis developed, and she subsequently died. Invasive lobular carcinoma is characterized by the development of gastrointestinal metastases and is rarely detected before autopsy. We describe our experience with a patient in whom invasive lobular carcinoma of the breast with metastasis to the duodenal wall was definitively diagnosed on laparotomy. PMID- 18386119 TI - 64-slice computed tomography coronary angiography: diagnostic accuracy in the real world. AB - PURPOSE: This study was done to evaluate the diagnostic accuracy of 64-slice computed tomography coronary angiography (CTCA) for the detection of significant coronary artery stenosis in the real clinical world. MATERIALS AND METHOD: From the CTCA database of our institution, we enrolled 145 patients (92 men, 52 women, mean age 63.4 +/- 10.2 years) with suspected coronary artery disease. All patients presented with atypical or typical chest pain and underwent CTCA and conventional coronary angiography (CA). For the CTCA scan (Sensation 64, Siemens, Germany), we administered an IV bolus of 100 ml of iodinated contrast material (Iomeprol 400 mgI/ml, Bracco, Italy). The CTCA and CA reports used to evaluate diagnostic accuracy adopted > or =50% and > or =70%, respectively, as thresholds for significant stenosis. RESULT: Eleven patients were excluded from the analysis because of the nondiagnostic quality of CTCA. The prevalence of disease demonstrated at CA was 63% (84/134). Sensitivity, specificity and positive and negative predictive values for CTCA on a per-segment, per-vessel, and per-patient basis were 75.6%, 85.1%, 97.6%; 86.9%, 81.8%, 58.0%; 48.2%, 68.1%, 79.6%; and 95.7%, 92.3%, 93.5%, respectively. Only two out of 134 eligible patients were false negative. Heart rate did not significantly influence diagnostic accuracy, whereas the absence or minimal presence of coronary calcification improved diagnostic accuracy. The positive and negative likelihood ratios at the per patient level were 2.32 and 0.041, respectively. CONCLUSION: CTCA in the real clinical world shows a diagnostic performance lower than reported in previous validation studies. The excellent negative predictive value and negative likelihood ratio make CTCA a noninvasive gold standard for exclusion of significant coronary artery disease. PMID- 18386120 TI - Tracheal and main bronchial diverticula: the role of CT. AB - PURPOSE: This paper describes an asymptomatic and rarely diagnosed entity without a clear correlation with respiratory disease identified in 1%-2% of cases at spiral computed tomography (CT). MATERIALS AND METHODS: We retrospectively reviewed the CT features of air collections contiguous to the tracheal or main bronchial wall and communicating with the airway in 16 patients undergoing CT for other reasons. Four patients were also studied by virtual bronchoscopy and three by actual bronchoscopy. RESULTS: Thirteen of the 16 patients had a single diverticulum, whereas the remaining three had multiple diverticula. Only one patient had a bronchial diverticulum. The most frequent site was the right posterolateral wall of the trachea at the level of the second or third thoracic vertebral body. In 2/3 of patients, chronic obstructive pulmonary disease was not documented. CONCLUSIONS: CT was the most effective method for evaluating the presence and features of diverticula. A correlation between tracheobronchial diverticula and chronic respiratory disease was documented in 5/16 patients, whereas no respiratory disorder was identified in the remaining 11. Although chronic inflammatory tracheobronchial changes and increased endoluminal pressure may be important causes, we hypothesise that other, as yet unknown, aetiopathological factors could exist. PMID- 18386121 TI - Ultrasound diagnosis of pneumonia in children. AB - PURPOSE: This study was done to compare the diagnostic accuracy of ultrasound and chest X-ray (CXR) in children with suspected pneumonia. MATERIALS AND METHODS: Seventy-nine children aged from 6 months to 16 years with clinical signs suggestive of pneumonia underwent lung ultrasound and CXR. RESULTS: Lung ultrasound was positive for the diagnosis of pneumonia in 60 patients, whereas CXR was positive in 53. In four patients with negative CXR and positive ultrasound findings, pneumonia was confirmed by chest computed tomography (CT) (performed for recurrent pneumonia in the same location). In the other three patients with negative CXR and positive ultrasound findings, the clinical course was consistent with pneumonia. CONCLUSIONS: Lung ultrasound is a simple and reliable tool that can be used by the clinician in the case of suspected pneumonia. It is as reliable as CXR, can be easily repeated at the patient's bedside and carries no risk of irradiation. PMID- 18386122 TI - Malignant renal neoplasms: correlation between ADC values and cellularity in diffusion weighted magnetic resonance imaging at 3 T. AB - PURPOSE: This study aimed at exploring the feasibility of high-field diffusion weighted magnetic resonance imaging (DW-MRI) (3 T) and to correlate apparent diffusion coefficient (ADC) values with tumour cellularity in renal malignancies. MATERIALS AND METHODS: Thirty-seven patients (ten healthy volunteers and 27 patients with suspected renal malignancy) underwent T1-, T2-weighted and T1 weighted contrast-enhanced magnetic resonance imaging (MRI). Diffusion-weighted images were obtained with a single-shot spin-echo echo-planar imaging (SE-EPI) sequence with a b value of 500 s/mm(2). All lesions were surgically resected, and mean tumour cellularity was calculated. Comparison between tumour cellularity and mean ADC value was performed using simple linear regression analysis. RESULTS: The mean ADC value in normal renal parenchyma was 2.35+/-0.31 x 10(-3) mm(2)/s, whereas mean ADC value in renal malignancies was 1.72+/-0.21 x 10(-3) mm(2)/s. In our population, there were no statistically significant differences between ADC values of different histological types. The analysis of mean ADC values showed an inverse linear correlation with cellularity in renal malignancies (r=-0.73, p<0.01). CONCLUSIONS: DW-MRI is able to differentiate between normal and neoplastic renal parenchyma on the basis of tissue cellularity. PMID- 18386123 TI - Diffusion-weighted MRI sequences (DW-MRI) of the kidney: normal findings, influence of hydration state and repeatability of results. AB - PURPOSE: This study was performed to determine the apparent diffusion coefficient (ADC) of the normal kidney using diffusion-weighted magnetic resonance imaging (DW-MRI) sequences and to analyse both the changes due to hydration state and results repeatability. MATERIALS AND METHODS: Ten volunteers underwent DW-MRI imaging of the kidneys with a breath-hold single-shot spin-echo planar imaging (SE-EPI) sequence in the axial and coronal planes with b values of 300, 500, 800 s/mm(2), in different states of hydration. Urine osmolarity (OsmU) and sodium excretion (NaU) were measured at the time of each examination. ADC maps were created for all b values, and ADC values were calculated and compared between different states of hydration. In five subjects, the protocol was conducted twice to test data repeatability. RESULTS: ADC values were lower with higher b values (3.00 vs. 2.47 vs. 1.99 x 10(-3) mm(2)/s with b values of 300, 500, 800 s/mm(2), respectively). ADC values in different hydration states were not statistically different. Measurements were reproducible. OsmU and NaU were statistically different in the different states of hydration (p<0.01). CONCLUSIONS: ADC values significantly decrease with higher b values. Average ADC values in the normal kidney are reproducible. Hydration state does not significantly influence mean ADC values. PMID- 18386124 TI - Magnetic resonance imaging versus ultrasonography in fetal pathology. AB - PURPOSE: This paper describes our experience with magnetic resonance imaging (MRI) in the assessment of fetal anatomical structures and major fetal pathologies. MATERIALS AND METHODS: The retrospective study included 128 pregnant women between the 22nd and 38th week of gestation. We used the following imaging protocol: T2-weighted single-shot fast spin-echo sequences for all foetuses and, in selected cases, gradient echo with steady-state free precession (SSFP), T1 weighted spoiled gradient echo [fast low-angle shot (FLASH)] with and without fat saturation, and T2 thick-slab sequences with multiplanar technique. In 32 cases, we performed diffusion-weighted sequences with apparent diffusion coefficient (ACD) maps on the brain, the kidneys and the lungs. RESULTS: We achieved diagnostic-quality images in 125 of 128 patients; MR image quality was unsatisfactory in three cases only. In 16 cases with previous negative ultrasound (US) findings, MRI confirmed the US diagnosis. MRI confirmed the positive US diagnosis in 67 of 109 cases (61.5%); in 11 cases it changed the US diagnosis, and in 31/109 the examination was negative. In addition, MRI identified other anomalies not recognised during US examination. CONCLUSIONS: With its ultrafast sequences, fetal MRI provides good detail of normal fetal anatomy and allows characterisation of suspected anomalies. PMID- 18386125 TI - Role of color-Doppler sonography in the follow-up of renal artery stenting. AB - PURPOSE: The aim of this study was to evaluate the utility of color-Doppler ultrasound (CDUS) in detecting haemodynamically significant in-stent restenosis in patients who underwent endoluminal renal artery revascularisation by stent deployment. MATERIALS AND METHOD: Between January 2000 and December 2006, 42 patients (nine women and 33 men, age range 45-87 years) treated by endovascular renal artery stenting were studied with CDUS. Renal artery haemodynamics were evaluated to identify haemodynamically significant restenosis. Patients with CDUS signs of restenosis underwent angiography for a possible further revascularisation procedure. RESULT: Of 42 patients examined by CDUS, 13 (31%) showed signs of haemodynamically significant in-stent renal artery restenosis. Of these 13, three did not undergo endoluminal renal artery revascularisation because renal ischaemia deterioration and irreversible renal circulation impairment. In the remaining ten patients (23.8%), who had no signs of severe nephropathy, angiography confirmed the CDUS findings of in-stent restenosis in all cases. Restenoses were successfully treated by a repeat endovascular revascularisation procedure. CONCLUSION: Our results confirm the fundamental role of CDUS in the follow-up of patients after renal artery stenting. It enables early restenosis detection and evaluation of renovascular disease associated with renal artery stenosis. CDUS provides essential information for the subsequent clinical management of these patients. PMID- 18386126 TI - Role of breast magnetic resonance imaging (MRI) in patients with unilateral nipple discharge: preliminary study. AB - PURPOSE: This study was performed to assess the role of magnetic resonance imaging (MRI) in patients with unilateral nipple discharge. MATERIALS AND METHODS: Forty-four patients with bloody or serosanguineous nipple discharge and negative mammographic findings (35/44 cases) underwent MRI for evaluation of breast ducts. Ultrasonography, negative in 18 patients, identified 26 cases of ductal ectasia (12 simple, nine with solid intraductal echoes and wall thickening, five with inhomogeneous parenchyma). Galactography was negative in three patients and positive in nine. Nineteen patients were followed up by clinical examination, ultrasonography, and cytological evaluation of nipple discharge (6-12 months); three patients underwent excisional biopsy, ten core biopsy and 12 cytological biopsy (followed by excisional biopsy). RESULTS: MRI identified 25 enhancing lesions Breast Imaging Reporting and Data Systems (BI RADS) 3 or 4) and confirmed the galactographic findings (ductal ectasia, intraluminal filling defects). Five papillomatoses appeared as patchy, homogeneous enhancing areas, 15 intraductal papillomas as areas with well-defined margins and type II time-intensity curves, and two atypical ductal hyperplasias as diffuse nodular enhancement. One micropapillary ductal carcinoma in situ (DCIS), one papillary carcinoma and one infiltrating ductal carcinoma (IDC) were visualised as two segmental areas of enhancement and one mass-like enhancement with poorly defined margins (BI-RADS 4). The follow-up was negative, showing no pathological enhancement (BI-RADS 1) in 12 patients and benign enhancement (BI RADS 2) in seven. CONCLUSIONS: Breast MRI can be considered a valuable examination in the diagnosis of suspected ductal disease and an alternative to galactography when the latter cannot be used. PMID- 18386127 TI - Bionic ear imaging. AB - PURPOSE: The aim of this study was to illustrate the different imaging features of middle and inner ear implants, brainstem implants and inferior colliculus implants. MATERIALS AND METHODS: We retrospectively reviewed the computed tomography (CT) images of 468 patients with congenital or acquired transmissive or neurosensory hearing loss who underwent surgery. The implants examined were: 22 Vibrant Soundbridge implants, 5 at the long limb of the incus and 17 at the round window, 350 cochlear implants, 95 brainstem implants and 1 implant at the inferior colliculus. All patients underwent a postoperative CT scan (single or multislice scanner) and/or a Dentomaxillofacial cone-beam CT scan (CBCT) (axial and multiplanar reconstruction), and/or a plain-film radiography to visualise the correct position of the implant. RESULTS: The CBCT scan depicts Vibrant site of implant better than plain-film radiography, with a lower radiation dose compared to CT. For cochlear implants, a single plain radiograph in the Stenvers projection can directly visualise the electrodes in the cochlea. All patients with brainstem or inferior colliculus implants underwent postoperative CT to exclude complications and the assess correct implantation, but the follow-up of these implants can be performed by plain radiography alone. CONCLUSIONS: CT and CBCT scans are reliable and relatively fast methods for precisely determining the location of middle ear implants. CBCT is preferable to CT because of the lower radiation dose administered; a single plain-film radiograph is enough to visualise and follow-up cochlear, brainstem and inferior colliculus implants. PMID- 18386128 TI - Postsurgical diagnostic evaluation of patients with differentiated thyroid carcinoma: comparison of ultrasound, iodine-131 scintigraphy and PET with fluorine-18 fluorodeoxyglucose. AB - PURPOSE: The aim of this study was to compare the results of ultrasound (US), whole-body scintigraphy with iodine-131 (I-131 WBS) and positron emission tomography with fluorine-18 deoxyglucose (FDG-PET) in the follow-up of patients after thyroidectomy for differentiated thyroid carcinoma (DTC). MATERIALS AND METHODS: Thirteen patients (3 men, 10 women) were evaluated by neck US, I-131 WBS and FDG-PET. In each patient six anatomical regions (right and left thyroid bed, right and left cervical region, right and left supraclavicular region) were investigated, for a total of 78 regions. Distant metastases were investigated by I-131 WBS and FDG-PET and considered separately in the analysis. Imaging findings were compared with the reference standards, such as fine-needle aspiration cytology (2), biopsy (4) or clinical-radiological studies (7). RESULTS: US, FDG PET and I-131 WBS showed concordant negative results in most (70, 90%) of the anatomical sites considered. In one patient with left cervical lymph node metastasis, the imaging techniques showed concordant positive results (1%). In the remaining 7 regions (9%), the imaging results were discordant; in particular, tumour lesions, nodal metastases (4) and thyroid bed recurrences (3) were detected by US only (3), by US and I-131 WBS (1) and by FDG-PET only (3). With regard to distant metastases, FDG-PET and I-131 WBS yielded concordant negative results in the majority (77%) of patients (9); in one patient only were the two imaging techniques concordant in their positive result. In the last three patients, the results were discordant; in particular, distant metastases were detected by I-131 WBS only in two patients and by FDG-PET only in one patient. CONCLUSIONS: Our work indicates a fundamental role for US in evaluation of the neck after surgery for DTC. WBS is useful to determine differentiation of tumour lesions, to identify thyroid remnants and to look for distant metastases. FDG-PET has an important role in cases of dedifferentiated thyroid carcinoma in which WBS and thyroglobulin measurements are unable to detect tumour lesions. PMID- 18386129 TI - In vivo, high-field, 3-Tesla 1H MR spectroscopic assessment of liver fibrosis in HCV-correlated chronic liver disease. AB - PURPOSE: Histology is the gold standard by which to diagnose and score hepatic fibrosis. Recently, it has been proposed that hepatic magnetic resonance spectroscopy (MRS) could provide an accurate representation of the disease process. The aim of this study was to correlate the in vivo high-field (3-Tesla) (1)H MRS features of noncirrhotic chronic hepatitis C patients stratified according to the histopathological stages of fibrosis. MATERIALS AND METHOD: Six healthy controls and 23 patients with biopsy-proven precirrhotic hepatitic C virus (HCV)-related liver disease were included. The subdivision of patients into the histopathological stages of fibrosis was based on the Ishak fibrosis (F) scoring system: mild hepatitis (0< or =F< or =1), moderate (2< or =F< or =3) and severe hepatitis (4< or =F< or =5). For correlation analysis, the Spearman nonparametric test was used. Differences between groups were calculated with the nonparametric Mann-Whitney U test. A p value <0.05 was considered significant. The particular metabolite content was evaluated in relative units (RU), according to the pattern metabolite/H(2)O=area of the metabolite x1,000/area of nonsuppressed water. RESULT: A significant statistical difference was observed between control vs. mild and moderate vs. severe disease severity in choline containing compounds (CCC)/H(2)O ratios (p=0.0379 and p=0.0003) and in glutamine/glutamate (Glx)/H(2)O ratios (p=0.004 and p<0.0001), whereas a statistically significant difference in the lipid/H(2)O ratios was achieved only between control vs. moderate and between moderate vs. severe stages of disease (p=0.011 and p=0.0030). CONCLUSION: High-field (1)H MRS successfully differentiates between mild/moderate vs. severe stages of chronic hepatitis and can be considered a complement to most standard imaging protocols in the liver. PMID- 18386130 TI - Supervised automatic procedure to identify new lesions in brain MR longitudinal studies of patients with multiple sclerosis. AB - PURPOSE: Identification of new enhancing lesions is a major endpoint of longitudinal brain magnetic resonance (MR) studies of multiple sclerosis (MS). To date, this is a visual, time-consuming procedure. We present here a supervised automated procedure (SAP) aimed at reducing the time needed to identify new MS enhancing lesions. MATERIALS AND METHODS: The SAP uses an algorithm including Cartesian coordinates of the lesions to be compared, their area and a constant (k). The procedure was validated for enhancing lesions on T1-weighted spin-echo images after intravenous administration of 0.1 mmol/kg of paramagnetic contrast agent, randomly selected from a dataset of a longitudinal MR study on ten relapsing-remitting MS patients followed for 2-5 years. During the validation session, two readers decided by consensus whether two lesions, present on the same slice of two examinations performed on subsequent dates, were the same or not. In this way, k was calibrated to obtain the same result from both visual inspection and automatic algorithm output. RESULTS: After evaluating of 25+/-5 (mean+/-standard deviation) lesions in each of ten different sessions with correction of k value, the k value became a stable value (0.45+/-0.05). CONCLUSIONS: Once the suitable value of k was found, SAP was able to identify new enhancing lesions, avoiding visual inspection, which is usually a lengthy procedure. PMID- 18386131 TI - A BAC library of Beta vulgaris L. for the targeted isolation of centromeric DNA and molecular cytogenetics of Beta species. AB - We constructed a sugar beet (Beta vulgaris) bacterial artificial chromosome (BAC) library of the monosomic addition line PAT2. This chromosomal mutant carries a single additional chromosome fragment (minichromosome) derived from the wild beet Beta patellaris. Restriction analysis of the mutant line by pulsed-field gel electrophoresis was used to determine HindIII as a suitable enzyme for partial digestion of genomic DNA to generate large-insert fragments which were cloned into the vector pCC1. The library consists of 36,096 clones with an average insert size of 120 kb, and 2.2% of the clones contain mitochondrial or chloroplast DNA. Based on a haploid genome size of 758 Mbp, the library represents 5.7 genome equivalents providing the probability of 99.67% that any sequence of the PAT2 genome can be found in the library. Hybridization to high density filters was used to isolate 89 BACs containing arrays of the centromere associated satellite repeats pTS5 and pTS4.1. Using the identified BAC clones in fluorescent in situ hybridization experiments with PAT2 and Beta patellaris chromosome spreads their wild beet origin and centromeric localization was demonstrated. Multi-colour FISH with differently labelled satellite repeats pTS5 and pTS4.1 was used to investigate the large-scale organization of the centromere of the PAT2 minichromosome in detail. FISH studies showed that the centromeric satellite pTS5 is flanked on both sides by pTS4.1 arrays and the arms of the minichromosome are terminated by the Arabidopsis-type telomeric sequences. FISH with a BAC, selected from high-density filters after hybridization with an RFLP marker of the genetic linkage group I, demonstrated that it is feasible to correlate genetic linkage groups with chromosomes. Therefore, the PAT2 BAC library provides a useful tool for the characterization of Beta centromeres and a valuable resource for sugar beet genome analysis. PMID- 18386132 TI - The fate of exogenous iodine in pot soil cultivated with vegetables. AB - A pot experiment was conducted to explore a more effective approach to enhancing vegetable uptake of soil iodine, with the ultimate goal of using agricultural fortification as a measure to prevent iodine deficiency disorders in local communities. Two types of iodine fertilizers were added separately to pot soil samples at various dosages. The fortified soil in each of the flower pots was seeded with one of four test crops (pakchoi, celery, pepper, and radish) in an effort to examine the effect of vegetable cultivation. The fate and residual levels of the exogenous iodine in the fortified soil samples were then monitored and quantified. The data showed that the soil iodine contents decreased with time (and hence with plant growth as well). At the second cutting, iodine from the inorganic form (KI) as the exogenous source was reduced to approximately 50% (41.6-61.0%) of the applied dose, whereas that in soil fortified with the seaweed fertilizer was down to approximately 60% (53.9-71.5%). The abilities of the edible portion of the four vegetables in accumulating the soil iodine were as follows: pakchoi > celery > radish > pepper. On the whole, iodine residues were found less in soil cultivated with vegetables. Vegetable cultivation appeared to have enhanced the soil content of the water-soluble form of iodine somewhat, especially in soil fortified with the inorganic forms. There also appeared to be a significant negative correlation between the residual iodine and its dissolution rate in soil. Overall, the results of the present study pointed toward the direction that the seaweed fertilizer tends to be a (more) preferred source of agricultural fortification in promoting human iodine nutrition. PMID- 18386133 TI - The burden of diabetes-related mortality in France in 2002: an analysis using both underlying and multiple causes of death. AB - AIM: To describe the burden of diabetes-related mortality in France. METHODS: Underlying and multiple causes (all causes listed) of death were extracted from the 2002 French national mortality registry. Death rates were standardized on the age structure of the European population. RESULTS: Diabetes was reported as the underlying cause of death in 11,177 certificates (2.1%), and as multiple causes in 29,357 certificates (5.3%), giving a ratio (multiple/underlying causes) of 2.6. When diabetes was a multiple cause, the mean age at death was 75 years in men, 81 years in women. The age-standardized mortality rates were 41.0/100,000 in men, 24.6/100,000 in women. The excess mortality observed in men (men/women ratio = 1.7) decreased with age. Geographic differences were observed: higher rates in the North-East, lower rates in the West of the country. In certificates mentioning diabetes, the most frequent cause of death was diseases of the circulatory system (76%). Coronary heart diseases, foot ulcers and renal diseases were more likely to be mentioned in certificates referring to diabetes than in those that did not. DISCUSSION: The use of multiple rather than underlying causes of death more than doubled diabetes-related mortality rates. While probably still under-estimated, the burden of diabetes-related mortality corresponds to a high proportion of the total mortality, especially in men. Geographic differences partially reflect disparities in diabetes prevalence. Causes more frequently associated with diabetes include coronary heart disease and complications related to neuropathy and nephropathy. PMID- 18386134 TI - Characterisation of breast cancer cell lines and establishment of a novel isogenic subclone to study migration, invasion and tumourigenicity. AB - The process of tumour invasion and subsequent metastasis represents the most lethal aspect of cancer. In this study the invasive and migratory activity of four human breast cancer cell lines; MCF-7, MDA-MB-231, BT474 and Hs578T was investigated. Isogenic subclones were isolated from the Hs578T cell line using sequential passages through the BD Matrigel Invasion Chamber assay system. A new invasive subclone designated, Hs578Ts(i)8 was isolated and shown to be 3-fold more invasive and 2.5-fold more migratory than the parental cell line. The variant cells formed up to 25 times more colonies in soft agar and also produced tumours in vivo in nude mice. Flow cytometry analysis showed that the Hs578Ts(i)8 cells had 30% more CD44+/CD24-/low cells than the parental Hs578T cell line. The presence of a breast cancer stem cell population within the variant cell line may provide an explanation for the increased anchorage independent growth and tumourigenicity. PMID- 18386135 TI - Epigenetic contributions to cancer metastasis. AB - The molecular basis of cancer encompasses both genetic and epigenetic alterations. These epigenetic changes primarily involve global DNA methylation changes in the form of widespread loss of methylation along with concurrent hypermethylation events in gene regulatory regions that can repress tissue specific gene expression. Increasingly, the importance of these epigenetic changes to the metastatic process is being realized. Cells may acquire an epi genotype that permits their dissemination from the primary tumour mass or the ability to survive and proliferate at a secondary tissue site. These epigenetic changes may be cancer-type specific, or in some cases may involve a common target gene providing a selective advantage to multiple metastatic cell types. In this review, I examine the growing volume of literature related to the epigenetic contributions to cancer metastasis. I discuss the functional importance of these epigenetic phenomena and how new epigenetic biomarkers may permit the identification of diagnostic signatures of metastasis and the development of new cancer therapies. PMID- 18386136 TI - Influence of diet on metastasis and tumor dormancy. AB - Tumor metastasis is responsible for most cancer deaths, and can occur after long periods of tumor dormancy. Information learned from experimental studies on tumor metastasis and dormancy is shedding light on mechanisms responsible and possible therapeutic approaches. 'Seed' (the cancer cell) and 'soil' (the microenvironment of the secondary organ) factors contribute to metastatic outcome. This review considers the possibility that various dietary components may affect both 'seed' and 'soil' compartments, thereby influencing the growth of metastases, and discusses an experimental study on dietary genistein that illustrates this concept. While studies on human diet are complex, the possibility that relatively non-toxic dietary intervention strategies could impact on metastasis and patient survival is attractive and worthy of further study in appropriate experimental models of metastasis and tumor dormancy. PMID- 18386137 TI - Attenuation of cisplatin nephrotoxicity by inhibition of soluble epoxide hydrolase. AB - Cisplatin is a highly effective chemotherapeutic agent against many tumors; however, it is also a potent nephrotoxicant. Given that there have been no significant advances in our ability to clinically manage acute renal failure since the advent of dialysis, the development of novel strategies to ablate nephrotoxicity would represent a significant development. In this study, we investigated the ability of an inhibitor of soluble epoxide hydrolase (sEH), n butyl ester of 12-(3-adamantan-1-yl-ureiido)-dodecanoic acid (nbAUDA), to attenuate cisplatin-induced nephrotoxicity. nbAUDA is quickly converted to AUDA and results in maintenance of high AUDA levels in vivo. Subcutaneous administration of 40 mg/kg of nbAUDA to C3H mice every 24 h resulted in elevated blood levels of AUDA; this protocol was also associated with attenuation of nephrotoxicity induced by cisplatin (intraperitoneal injection) as assessed by BUN levels and histological evaluation of kidneys. This is the first report of the use of sEH inhibitors to protect against acute nephrotoxicity and suggests a therapeutic potential of these compounds. PMID- 18386138 TI - Morphological alterations induced by doxorubicin on H9c2 myoblasts: nuclear, mitochondrial, and cytoskeletal targets. AB - Doxorubicin (Dox) is a very potent antineoplastic agent used against several types of cancer, despite a cumulative cardiomyopathy that reduces the therapeutic index for treatment. H9c2 myoblast cells have been used as an in vitro model to study biochemical alterations induced by Dox treatment on cardiomyocyte cells. Despite the extensive work already published, few data are available regarding morphological alterations of H9c2 cells during Dox treatment. The purpose of the present work was to evaluate Dox-induced morphological alterations in H9c2 myoblasts, focusing especially on the nuclei, mitochondria, and structural fibrous proteins. Treatment of H9c2 cell with low concentrations of Dox causes alterations in fibrous structural proteins including the nuclear lamina and sarcomeric cardiac myosin, as well as mitochondrial depolarization and fragmentation, membrane blebbing with cell shape changes, and phosphatidylserine externalization. For higher Dox concentrations, more profound alterations are evident, including nuclear swelling with disruption of nuclear membrane structure, mitochondrial swelling, and extensive cytoplasm vacuolization. The results obtained indicate that Dox causes morphological alterations in mitochondrial, nuclear, and fibrous protein structures in H9c2 cells, which are dependent on the drug concentration. Data obtained with the present study allow for a better characterization of the effects of Dox on H9c2 myoblasts, used as a model to study Dox-induced cardiotoxicity. The results obtained also provide new and previously unknown targets that can contribute to understand the mechanisms involved in the cardiotoxicity of Dox. PMID- 18386139 TI - Cancer risk in a cohort of patients with systemic lupus erythematosus (SLE) in California. AB - OBJECTIVE: We conducted a retrospective cohort study to examine cancer risk in a large cohort of systemic lupus erythematosus (SLE) patients in California. METHODS: The cohort consisted of individuals with SLE derived from statewide patient discharge data during the period 1991-2002. SLE patients were followed using cancer registry data to examine patterns of cancer development. Standardized incidence ratios (SIRs) and 95% CI were calculated to compare the observed to expected numbers of cancers based on age-, race-, and sex-specific incidence rates in the California population. RESULTS: The 30,478 SLE patients were observed for 157,969 person-years. A total of 1,273 cancers occurred within the observation interval. Overall cancer risk was significantly elevated (SIR = 1.14, 95% CI = 1.07-1.20). SLE patients had higher risks of vagina/vulva (SIR = 3.27, 95% CI = 2.41-4.31) and liver cancers (SIR = 2.70, 95% CI = 1.54-4.24). Elevated risks of lung, kidney, and thyroid cancers and several hematopoietic malignancies were also observed. Individuals had significantly lower risks of several screenable cancers, including breast, cervix, and prostate. CONCLUSIONS: These data suggest that risks of several cancer types are elevated among SLE patients. Detailed studies of endogenous and exogenous factors that drive these associations are needed. PMID- 18386140 TI - Deaths from non-melanoma skin cancer in Western Australia. AB - OBJECTIVES: Non-melanoma skin cancer (NMSC) is common, slow growing, and rarely metastasizes. However, there are still nearly 400 deaths from NMSC in Australia annually. We aimed to investigate the accuracy of NMSC death coding and to describe the characteristics of these deaths and the potential for prevention. METHODS: Histology reports for all deaths coded as NMSC (ICD-10 C44.0-C44.9) by the Western Australian Cancer Registry for the years 1996-2005 were reviewed for type of cancer, body site (primary tumor and metastases), and level of available documentation. RESULTS: Of 368 deaths recorded as being due to NMSC only 3 were found to be miscoded. An additional 53 deaths contained inadequate information to confirm NMSC as the cause of death. Of the confirmed cases, 219 were due to squamous cell carcinoma, 53 to Merkel cell carcinomas, and 40 to other skin cancers. Cases were mainly males and were elderly. Most of the primary squamous and Merkel cell carcinomas were in areas of maximum sun exposure (face, ears, and hands, and scalp in males). CONCLUSIONS: Misclassification of NMSC deaths in WA was minimal. The majority of NMSC deaths were due to squamous cell carcinomas; had primary sites associated with significant sun exposure; and occurred in older men. PMID- 18386141 TI - Dietary factors and risk of t(14;18)-defined subgroups of non-Hodgkin lymphoma. AB - OBJECTIVE: To evaluate the associations between diet and non-Hodgkin lymphoma (NHL) according to t(14;18) status, one of the most common chromosomal abnormalities in NHL, as t(14;18)-positive NHL represents a genetically more homogeneous group than NHL overall. METHODS: We determined the presence of the t(14;18)(q32;q21) by fluorescence in situ hybridization in 172 of 175 tumor blocks from a population-based, case-control study conducted in Nebraska during 1983-1986. Information on the frequency of consumption as an adult of 30 food items was derived from the parent case-control study. Dietary factors in 60 t(14;18)-positive and 87 t(14;18)-negative cases were compared with 1,075 controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using polytomous logistic regression. RESULTS: The risk of t(14;18)-positive NHL for the highest versus the lowest approximate tertile of intake was elevated for milk (OR = 2.2; 1.0-5.0) and dietary nitrite (OR = 2.8; 1.3-6.1), whereas coffee consumption was inversely associated with risk (OR = 0.4; 0.2-0.7). We also found inverse associations between the intake of fish (OR = 0.5; 0.3-1.0) and carotene (OR = 0.5; 0.2-0.9) and risk of t(14;18)-negative NHL. There was no association between the intake of meats, vegetables, protein, or vitamin C and risk of either t(14;18)-positive or t(14;18)-negative NHL. CONCLUSION: We observed differences in associations between diet and t(14;18)-defined subgroups of NHL. These findings should be interpreted cautiously because of the small sample. PMID- 18386142 TI - Novel function of the thyroid hormone analog tetraiodothyroacetic acid: a cancer chemosensitizing and anti-cancer agent. AB - Previous studies from our laboratory have demonstrated that thyroid hormones play a key role in cancer progression. In addition, a deaminated form, tetraiodothyroacetic acid (tetrac), that antagonizes the proliferative action of these hormones was found to possess anti-cancer functions through its ability to inhibit cellular proliferation and angiogenesis. The present study was undertaken to investigate whether tetrac could also suppress the development of drug resistance, known as a causative factor of disease relapse. Tetrac was shown to enhance cellular response in vitro to doxorubicin, etoposide, cisplatin, and trichostatin A in resistant tumor cell lines derived from neuroblastoma, osteosarcoma, and breast cancer. The mechanism of action of tetrac did not involve expression of classical drug resistance genes. However, radiolabeled doxorubicin uptake in cells was enhanced by tetrac, suggesting that one or more export mechanisms for chemotherapeutic agents are inhibited. Tetrac was also found to enhance cellular susceptibility to senescence and apoptosis, suggesting that the agent may target multiple drug resistance mechanisms. Tetrac has previously been shown to inhibit tumor cell proliferation in vitro. In vivo studies reported here revealed that tetrac in a pulsed-dose regimen was effective in suppressing the growth of a doxorubicin-resistant human breast tumor in the nude mouse. In this paradigm, doxorubicin-sensitivity was not restored, indicating that (1) the in vitro restoration of drug sensitivity by tetrac may not correlate with in vivo resistance phenomena and (2) tetrac is an effective chemotherapeutic agent in doxorubicin-resistant cells. PMID- 18386143 TI - Study of the voltammetric behaviour of the ethalfluralin and methalpropalin and its determination in environmental matrices at hanging mercury drop electrode. AB - An electroanalytical method has been developed for the determination of the herbicides ethalfluralin[N-ethyl-N-(2-methyl-2-propenyl)-2,6-dinitro-4 (trifluoromethyl) bezenamine] and methalpropalin [N-(2-methyl-2-propenyl)-2, 6 dinitro-N-propyl-4 (trifluoromethyl) benzenamine] by differential pulse adsorptive stripping voltammetry (DP-AdSV) on a hanging mercury drop electrode (HMDE) with universal buffer as supporting electrolyte. The optimum adsorption conditions were found to be pH 6.0, an accumulation potential of -0.6 V (HMDE vs SCE), an accumulation time of 80 s. and scan rate 45 mVs(-1). Calibration curve is linear in the range 1.30 x 10(-9) to 1.32 x 10(-5) M of ethalfluralin and 1.13 x 10(-5) to 2.0 x 10(-8) M of methalpropalin with detection limits of 1.08 x 10( 9) and 1.87 x 10(-8) M, respectively. The relative SD and correlation coefficients were found to be 1.24%, 0.998 and 1.34%, 0.995, respectively for ten replicates. The method is applied to the determination of the ethalfluralin and methalpropalin in formulations and environmental matrices. PMID- 18386144 TI - A simple method for simultaneous determination of some organic liquids in in process materials and effluents produced in the manufacture of amoxicillin and ampicillin. AB - A simple GC method for simultaneous determination of pivaloylchloride, methylacetoacetate, ethylacetoacetate and 2-ethylhexanoic acid in the presence of each other has been developed using glass column packed with 1% Silicone OV-17 on Gaschrom Q 100-120 mesh with temperature programming. Various performance parameters including precision, linearity and limit of detection have been evaluated. The method was found to be suitable for the analysis of these chemicals in in-process materials and effluents associated with the manufacture of amoxicillin and ampicillin. The run time was less than 15 min. The method has been successfully applied to determine the level of these hazardous organic liquids in real time samples. PMID- 18386145 TI - Spatial and temporal distribution of tributyltin (TBT) in seawater, sediments and bivalves from coastal areas of Korea during 2001-2005. AB - Tributyltin (TBT) concentrations were determined in seawater, sediments and bivalve samples collected from Korean coastal areas during 2001-2005, to investigate the levels and temporal variation in TBT contamination in relation to the timing of the imposition of regulations on TBT use in Korea. TBT concentrations ranged from <5.0 to 164 ng/L in seawater, from <7.0 to 9,576 ng/g dry weight in sediments, and from <7.0 to 6,296 ng/g dry weight in bivalves. The highest concentrations of TBT were found at locations close to intensive shipping traffic and industrial complexes, and the contamination at some hot spot areas was high enough to cause harmful effects on marine organisms. TBT concentrations and their occurrence in Korean coastal waters have been decreasing annually. In particular, TBT concentrations in seawater have dramatically decreased. This result is consistent with regulations and bans on the use of TBT in Korea. PMID- 18386146 TI - Concentration, distribution and source apportionment of atmospheric polycyclic aromatic hydrocarbons in the southeast suburb of Beijing, China. AB - Total suspended particle samples and gas phase samples were collected at three representative sampling sites in the southeastern suburb of Beijing from March 2005 to January 2006. The samples were analyzed for 16 US EPA priority PAHs using GC/MS. Concentrations of Sigma PAHs in particle and gas phases were 0.21-1.18 x 10(3) ng m(-3) and 9.5 x 10(2) ng-1.03 x 10(5) ng m(-3), respectively. PAH concentrations displayed seasonal variation in the order of winter>spring>autumn>summer for particle phase, and winter>autumn>summer>spring for gas phase. Partial correlation analysis indicates that PAH concentrations in particle phase are negatively correlated with temperature and positively correlated with air pollution index of SO(2). No significant correlation is observed between gas phase PAHs and the auxiliary parameters. Sources of PAH are identified through principal component analysis, and source contributions are estimated through multiple linear regression. Major sources of atmospheric PAHs in the study area include coal combustion, coke industry, vehicular emission and natural gas combustion. PMID- 18386147 TI - Analysis of the land use structure changes based on Lorenz curves. AB - Land use change has become a new field in global change research, and it is closely related to land resource security and sustainable development. The environmental simulation model was mostly used to study the land-use change before. In this paper, we use the Economic Lorenz curve and Gini coefficient of economic model to study the land use structure changes in the western plain of Jilin Province, China. The results showed that the Lorenz curve has the phenomenon of "two near and two far" and Gini coefficient "two increased and two decreased". That is to say, during the 15 years from 1989 to 2004, the areas of cultivated land and unused land had increased, Lorenz curves were near to the perfect equality line, and the distribution tended to be uniform; the areas of grassland and water body were reduced, the Lorenz curves were away from the perfect equality line, and the distribution tended to be increasingly uneven, which reflects the main process of the land use structure changes. Studying on the land use changes based on Economic model is simpler and more intuitionistic, and had the generalization values. PMID- 18386148 TI - SEM-EDX analysis of various sizes aerosols in Delhi India. AB - Scanning electron microscopy-energy dispersive X-ray analysis (SEM-EDX) was used to understand the differences in morphology, elemental composition and particle density of aerosols in different five size ranges to further investigate the potential sources as well as transport of pollutants from/at a much polluted and a very clean area of Delhi. Aerosol samples were obtained in five different size ranges viz. > or = 10.9, 10.9-5.4, 5.4-1.6, 1.6-0.7 and < or = 0.7 microm from a considerably very clean and a much polluted area of Delhi. It was observed that at polluted area most of the particles irrespective of size are of anthropogenic origin. At clean area, in coarse size fractions particles are of natural origin while in fine size range the presence of anthropogenic particles suggests the transport of particles from one area to the other. PMID- 18386149 TI - Biomonitoring acidic drainage impact in a complex setting using periphyton. AB - Acid mine drainage (AMD) often exerts various environmental pressures on nearby water courses: chemical stress from low pH and dissolved metals; physical stress from metal oxide deposits. Affected streams can thus display a spatially variable combination of stress agents that may complicate its biomonitoring using native communities such as periphyton. Here, we have measured water and periphyton variables in four streams that surround an abandoned copper mine to determine which periphyton attributes consistently detected AMD impact in a complex environmental setting. Seventeen years after the end of commercial exploitation, the abandoned mine still decreases water quality in nearby streams: moderate acidification, very high metal load (Al, Ni, Cu, Zn), and a conspicuous presence of metal oxide deposits with diverse composition. Even under the resultant complex pattern of polluted conditions, periphyton was a reliable bioindicator of AMD. Epilithic diatom taxa tolerant of acidic conditions increased in AMD sites and, at severely impacted locations, species richness decreased. Also, algal biomass may have been negatively affected in some stream reaches affected by metal oxide deposits. Other periphyton attributes (total biomass, diatom diversity) seemed mostly unrelated to AMD. Diatom assemblage composition was the most sensitive and consistent bioindicator of mine drainage; besides, it rendered a biological assessment of AMD impact that largely coincided with the physicochemical evaluation. Still, including other taxonomic (proportion of acid tolerant diatom species, diatom richness) and non-taxonomic (algal biomass) attributes in the biomonitoring procedure rendered a more comprehensive assessment of the negative consequences generated by AMD. PMID- 18386150 TI - Measurements of benzene and formaldehyde in a medium sized urban environment. Indoor/outdoor health risk implications on special population groups. AB - In the present study, the results of a measurement campaign aiming to assess cancer risk among two special groups of population: policemen and laboratory technicians exposed to the toxic substances, benzene and formaldehyde are presented. The exposure is compared to general population risk. The results show that policemen working outdoor (traffic regulation, patrol on foot or in vehicles, etc.) are exposed at a significantly higher benzene concentration (3-5 times) than the general population, while the exposure to carbonyls is in general lower. The laboratory technicians appear to be highly exposed to formaldehyde while no significant variation of benzene exposure in comparison to the general population is recorded. The assessment revealed that laboratory technicians and policemen run a 20% and 1% higher cancer risk respectively compared to the general population. Indoor working place air quality is more significant in assessing cancer risk in these two categories of professionals, due to the higher Inhalation Unit Risk (IUR) of formaldehyde compared to benzene. Since the origin of the danger to laboratory technicians is clear (use of chemicals necessary for the experiments), in policemen the presence of carbonyls in indoor air concentrations due to smoking or used materials constitute a danger equal to the exposure to traffic originated air pollutants. PMID- 18386151 TI - Monitoring of pesticide residues in vegetarian diet. AB - Samples (28) of complete vegetarian diet consumed from morning till night i.e. tea, milk, breakfast, lunch, snacks, dinner, sweet dish etc. were collected from homes, hostels and hotels periodically from Hisar and analysed for detecting the residues of organochlorine, synthetic pyrethriod, organophosphate and carbamate insecticides. The estimation was carried out by using multi-residue analytical technique employing gas chromatograph (GC)-electron capture detector and GC nitrogen phosphorous detector systems equipped with capillary columns. The whole diet sample was macerated in a mixer grinder and a representative sample in duplicate was analyzed for residues keeping the average daily diet of an adult to be 1,300 g. On comparing the data, it was found that actual daily intake (microgram/person/day) of lindane in two and endosulfan in four samples exceeded the acceptable daily intake. Residues of other pesticides in all the diet samples were lower than the acceptable daily intake (ADI) of the respective pesticides. The study concluded that although all the diet samples were found contaminated with one or the other pesticide, the actual daily intake of only a few pesticides was higher than their respective ADI. More extensive study covering other localities of Haryana has been suggested to know the overall scenario of contamination of vegetarian diet. PMID- 18386152 TI - Oral assessment and postgraduate medical examinations: establishing conditions for validity, reliability and fairness. AB - The purpose of this review was to examine the practice of oral assessment in postgraduate medical education in the context of the core assessment constructs of validity, reliability and fairness. Although oral assessment has a long history in the certification process of medical specialists and is a well established part of such proceedings for a wide range of specialties in most countries, there remains concern regarding its use. Therefore, there has been some move away from oral assessment for postgraduate medical education in some countries. This review also highlights the complexity of oral assessment as an examination format, partly through a consideration of the six dimensions of oral assessment, and raises concerns about the validity, reliability and fairness of such an assessment procedure for the award of certification of completion of the specialist training. Supporting high quality published research into examination practices and outcomes and acting on the findings of such research is needed urgently to allay concerns about the transparency and fairness of these examinations, especially when assessing international medical graduates. The article concludes by proposing 15 conditions under which oral assessment is valid, reliable and fair. PMID- 18386154 TI - Estradiol valerate and tibolone: effects upon brain oxidative stress and blood biochemistry during aging in female rats. AB - Estrogen compounds have been described as important brain protectors. This study investigated the effects of estradiol valerate (EV--0.3 mg/kg) and two concentrations of tibolone (TB1=0.5 mg/kg and TB2=1 mg/kg) on brain oxidative stress parameters and blood biochemistry in ovariectomized female rats, of three different age groups (young--2 months, adult--8 months, and old--20 months). In the brain cortex, young and old TB2-treated and old no-hormone-replacement (NR) females showed lower lipid hydroperoxide (LPO) levels compared to young Sham and adult TB1 animals (P<0.05). Also in the cortex, both tibolone doses produced higher (P<0.05) total antioxidant capacity (TOSC) levels compared to EV-treated adult females. Ovariectomized adult females (NR, EV, TB1 and TB2) showed lower (P<0.05) TOSC levels in the hippocampus compared to the Sham control. Reactive oxygen species (ROS) were higher (P<0.05) in old females compared to all younger ones. TB2-treated adults showed higher plasma glucose (P<0.05) levels compared to old animals. Regardless of age, TB2 treatment increased female (P<0.05) LDL levels compared to Sham and EV-treated animals. In old females, TB2 significantly increased HDL levels compared to Sham controls, and decreased triglyceride levels were shown in EV, TB1 and TB2 compared to Sham old females. The Atherogenic Index of Plasma was higher (P<0.05) in adult tibolone-treated females compared to both young and old TB2-treated females. These results suggest that the effects of gonad steroid on brain and blood physiology change significantly with aging, and that evaluating hormonal treatment types and doses could be the key factor in the potential use of a specific hormone therapy. PMID- 18386153 TI - Improvement in eGFR in patients with chronic kidney disease attending a nephrology clinic. AB - BACKGROUND: The adverse effects arising from late referral to a nephrologist of patients with chronic kidney disease (CKD) are well known. Retrospectively we examined the initial characteristics of patients referred in various stages of CKD to our nephrology division and tried to identify potential baseline factors associated with subsequent changes in estimated glomerular filtration rate (eGFR). PATIENTS AND METHODS: Between September 1997 and June 2006 1,443 patients (909 male, 534 female) with CKD, with eGFRs ranging from 15 to 89 ml/min, were referred to our nephrology division and categorized using the National Kidney Foundation classification for CKD based on eGFR. The slope of eGFR change (ml/min 1/1.73/m2-1/year-1) was determined by linear regression analysis and the patients were divided into five groups: (1) significantly progressive slope (deterioration) (more negative than -5 ml/min/year); (2) mildly progressive slope (>-5 to -1 to +1 to or=+5). RESULTS: At the first nephrology referral, 5.8% of the patients were on CKD stage 2 (eGFR: 90-60 ml/m), 46.7% on CKD stage 3 (eGFR: 59-30 ml/m), and 47.5% on CKD stage 4 (eGFR: 29-15 ml/m) CKD. Significantly improved slope was detected in 48.2% of CKD stage 2 patients, 29.3% of CKD stage 3 patients, and only 14.7% of CKD stage 4 patients (P<0.05). Being in stage 4 or stage 3 versus being in stage 2 significantly reduced the likelihood of an improved slope in logistic regression analysis whereas age, gender, presence of hypertension, and diabetes mellitus did not reach the level of significance. CONCLUSION: Referral to a nephrology clinic can lead not only to arrest of progression of CKD but also to regression/improvement. Early referral is a positive predictive factor for improvement in eGFR, which emphasizes the importance of such referral. The previously held idea that, once established, CKD progresses invariably is not valid anymore. PMID- 18386155 TI - Interaction between methotrexate and omeprazole in an adolescent with leukemia: a case report. AB - We present a case of severe mucositis due to a drug-drug interaction between methotrexate (MTX) and omeprazole in an adolescent with Acute Lymphatic Leukemia (ALL). In view of the prevalence of MTX in many cytotoxic protocols, and the frequent use of omeprazole in the (ambulatory) oncology setting, we believe that this case-report is worth mentioning. PMID- 18386156 TI - Abiogenic photophosphorylation of ADP to ATP sensitized by flavoproteinoid microspheres. AB - A model for abiogenic photophosphorylation of ADP by orthophosphate to yield ATP was studied. The model is based on the photochemical activity of flavoproteinoid microspheres that are formed by aggregation in an aqueous medium of products of thermal condensation of a glutamic acid, glycine and lysine mixture (8:3:1) and contain, along with amino acid polymers (proteinoids), abiogenic isoalloxazine (flavin) pigments. Irradiation of aqueous suspensions of microspheres with blue visible light or ultraviolet in the presence of ADP and orthophosphate resulted in ATP formation. The yield of ATP in aerated suspensions was 10-20% per one mol of starting ADP. Deaeration reduced the photophosphorylating activity of microspheres five to 10 times. Treatment of aerated microsphere suspensions with superoxide dismutase during irradiation partially suppressed ATP formation. Deaerated microspheres restored completely their photophosphorylating activity after addition of hydrogen peroxide to the suspension. The photophosphorylating activity of deaerated suspensions of flavoproteinoid microspheres was also recovered by introduction of Fe3+-cytochrome c, an electron acceptor alternative to oxygen. On the basis of the results obtained, a chemical mechanism of phosphorylation is proposed in which the free radical form of reduced flavin sensitizer (F1H*) and ADP are involved. PMID- 18386157 TI - A 25-kDa serine peptidase with keratinolytic activity secreted by Coccidioides immitis. AB - Coccidioides immitis is the causative agent of coccidioidomycosis, a systemic mycosis that attacks humans and a wide variety of animals. In the present study, we showed that the C. immitis mycelial form is able to release proteolytic enzyme into the extracellular environment. Under chemically defined growth conditions, mycelia secreted seven distinct polypeptides ranging from 15 to 65 kDa and an extracellular peptidase of 25 kDa. This enzyme had its activity fully inhibited by phenylmethylsulphonyl fluoride, a serine peptidase inhibitor. Conversely, metallo, cysteine, and aspartyl peptidase inhibitors did not alter the 25-kDa enzyme behavior. This extracellular serine peptidase was able to degrade keratin, a fibrous protein that composes human epidermis. Additionally, this peptidase cleaved different protein substrates, including gelatin, casein, hemoglobin, and albumin. Curiously, an 18-kDa serine peptidase activity was evidenced solely when casein was used as the co-polymerized protein substrate into the gel. The existence of different secreted peptidases could be advantageous for the adaptation of C. immitis to distinct environments during its complex life cycle. PMID- 18386158 TI - Malassezia folliculitis in China. AB - Superficial Malassezia folliculitis was diagnosed in 1-1.5% of all dermatology patients seen in a Dermatology Out-Patient clinic in Urumqi City, Xinjiang province, west China. It was most prevalent in healthy, middle-aged males in the warm seasons. This disease, first reported in 1969, is usually diagnosed as a systemic disease of immuno-compromised patients. Our findings indicate it is a relatively common superficial skin disease that can be managed with ketoconazole or itraconazole. PMID- 18386159 TI - Marasmius oreades substances block NF-kappaB activity through interference with IKK activation pathway. AB - The activation pathway of nuclear transcription factor kappa B (NF-kappaB) is a key mechanism for the progression of carcinogenesis at the molecular level. NF kappaB is related to the promotion of cell proliferation, inhibition of apoptosis, and the enhancement of tumor metastasis and angiogenesis. Marasmius oreades culture liquid extract, which was previously shown to affect NF-kappaB activation through inhibition of the phosphorylation of the inhibitory protein kappa B (IkappaBalpha), was subjected to liquid chromatography in order to investigate the specific mechanism of action of the active moieties present in the extract. Four fractions were obtained and tested for their abilities to block NF-kappaB activation pathway at different molecular levels. All fractions showed an anti-proliferative potential with no apparent cytotoxicity on MCF7 breast cancer cell line. Two out of the four fractions strongly affected the phosphorylation of IkappaBalpha and the NF-kappaB reporter activity in MCF7 breast cancer cell line. In addition, these two fractions prevented the p65 nuclear translocation and seemed to interfere with the IkappaB kinase (IKK) activation pathway. The IKK pathway is a major cellular signaling pathway set at a crossroad leading to NF-kappaB activation by a variety of stimuli. Also, these two fractions induced apoptosis of MCF7 cells. This study adds additional valuable data to our knowledge on the anticancer potential of fungal metabolites. It is the first report showing the medicinal value of M. oreades as a natural source of low-molecular-weight bioactive substances able to affect the process of tumorogenesis through the direct blockage of NF-kappaB activation at the IKK level. PMID- 18386160 TI - The regulation of TACE catalytic function by its prodomain. AB - AIM: To study the function of the prodomain of ADAM17 (TACE) and to develop an approach for interfering with inflammation processes. METHOD: The expression plasmids of the TACE ectodomain (T1300), prodomain (T591), signal peptide and prodomain (T648), full length (T2472), and the turncated TACE without prodomain (T57-T1824) were constructed and designated as pET-28a-T300, pET-28a-T591, pIRES2 EGFP-648, pEGFP-N1-T648, pIRES2-EGFP-T2472, and pIRES2-EGFP-T57-T1824, respectively. After Ni(2+)-NTA resin-affinity chromatography, the recombinant T591 and T1300 proteins were obtained and assayed by western blotting and circular dichroism. The experiment was carried out on THP1 cell lines stimulated by LPS in vitro. The inhibition of recombinant protein T591 to TACE activity was detected by ELISA and immunohistochemical detection. The expression plasmids (pIRES2-EGFP-T648, pIRES2-EGFP-T2472, and pIRES2-EGFP-T57-T1824) were used to transfect the U937 cells. HeLa cells were also transfected with pEGFP-N1-T648. The transfected U937 cells were then stimulated by LPS and the effect of expression plasmids on TNF-alpha secretion was detected by ELISA and flow cytometry (FCM). RESULTS: The recombinant prodomain protein inhibited 57% of the TNF-alpha secretion and mediated an accumulation of TNF-alpha on the surface of THP1 cells. An intense green fluorescence was seen in the membranes of HeLa cells transfected with pEGFP-N1-T648. The plasmid pIRES2-EGFP-T648 inhibited TNF-alpha secretion by 61.09% and mediated an accumulation of mTNF-alpha on the surface of the U937 cells. The secretion of sTNF-alpha and the level of the mTNF-alpha in the pIRES2-EGFP-T57-T1824 transfected cells gave no difference when compared with the pIRES2-EGFP transfected cells. Also the secretion of sTNF-alpha from the cells transfected by the plasmid pIRES2-EGFP-T2472 increased, while the level of mTNF-alpha decreased, compared with the pIRES2-EGFP-transfected cells. CONCLUSION: The prodomain has dual effects and might be useful in the molecular design of an anti-inflammatory drug. PMID- 18386161 TI - Identifying response shift statistically at the individual level. AB - OBJECTIVE: The purpose of this study was to explore whether a longitudinal comparison between reported and predicted health could be used as a method of identifying subjects who potentially experienced response shift. METHODS: A response-shift model was developed using data from a longitudinal study of stroke in which measures of stroke impact were made at study entry and at 1, 3, 6, and 12 months post stroke. Residuals from a random effects model were centered and used to create trajectories. This model was tested against a data set from a study in which the then-test had been administered. Twenty simulated data sets were also generated to examine how much of response shift could be attributed to random error. RESULTS: Group-based trajectory analysis identified seven trajectory groups. The majority (67%) of the 387 persons showed no response shift over time, whereas 15% lowered and 13% raised their health over time, disproportionally to that predicted. CONCLUSION: Results of the validation studies were supportive that this methodology identifies response shift, but further research is required to compare results with other methodologies and other predictive models. PMID- 18386162 TI - Significance of heat-shock protein (HSP) 90 expression in acute myeloid leukemia cells. AB - The 90-kDa heat shock protein (HSP90) is implicated in the conformational maturation and stabilization of a variety of client proteins with receptor and signal transduction functions. The objective of this study was to assess its expression in primary acute myeloid leukemia (AML) cells and to evaluate its biological and clinical significance. The in vitro effects of 17-AAG, a selective inhibitor of HSP90, was also evaluated. Cells from 65 patients with newly diagnosed AML were studied. The expression of HSP90 correlated with that of CD34, p170, and bcl-2 proteins but not with white cell counts, FAB or WHO subtype, or cytogenetics. HSP90 levels were also higher in samples exhibiting an autonomous growth in liquid culture or forming spontaneous colonies. A concomitant constitutive activation of the extracellular signal-regulated kinase and phosphatidylinositol 3-kinase/AKT pathways was observed in a majority of samples and was significantly correlated with HSP90 expression. All patients received induction chemotherapy. The percentages of HSP90-, CD34-, bcl-2-, and p170 positive cells were higher in patients who did not attain complete remission. Survival was also shorter in patients with high levels of HSP90. In vitro exposure of leukemic cells to 17-allylamino-demethoxy geldanamycin (17-AAG) resulted in inhibition of growth in liquid and clonogeneic cultures and in apoptosis, at concentrations which in most cases were not toxic for normal CD34 positive or progenitor cells. The concentration inhibiting 50% growth at 72 h in liquid culture correlated with HSP90 expression. Our study suggests that HSP90 is overexpressed in poor-prognosis AML cells and plays a role in cell survival and resistance to chemotherapy. Targeted therapy with 17-AAG represents a promising antileukemic strategy in adult AML. PMID- 18386163 TI - Different expression of occludin and ZO-1 in primary and metastatic liver tumors. AB - Tight junction (TJ) components were found to be correlated with carcinogenesis and tumor development. TJs are composed of three main integral membrane proteins; occludin, claudins and JAMs. Alteration of the TJ protein expression may play an important role in the process of cell dissociation, which is among the first steps of tumor invasion and metastasis. Reduced expression of ZO-1 has been reported to be associated with invasion of several tumors. The aim of the present study was to detect differences between occludin and ZO-1 expression in normal liver samples, HCCs and colorectal liver metastases. Expression of occludin and ZO-1 was analysed in 25 surgically removed human hepatocellular carcinomas (HCC) and 25 human colorectal liver metastases. Gene expression levels were measured by real-time RT PCR, protein expression was determined by immunohistochemistry, comparing tumors with the surrounding nontumorous parenchyma and with seven normal liver samples. Occludin and ZO-1 mRNAs showed significant downregulation in HCCs in comparison with normal liver and were also downregulated in the metastases when compared with normal liver. Occludin and ZO-1 proteins were weakly expressed on hepatocytes in normal liver, while strong expression was found on bile canaliculi. In HCCs occludin and ZO-1 did not show immunopositivity on tumor cells, while colorectal metastatic tumors revealed high levels of these molecules. HCCs and metastases are characterized by markedly different protein expression pattern of occludin and ZO-1, which phenomenon might be attributed to the different histogenesis of these tumors. PMID- 18386164 TI - Skip metastase on the left neck lymph nodes of the prostatic adenocarcinoma with neuroendocrine differentiation and accompanying thyroid micropapillary carcinoma. AB - We discuss here the thyroid micropapillary carcinoma that was detected incidentally when investigating the primary focus of the left neck multiple lymph node metastases occurring 8 months later in a patient of ours, whose pathological examination of radical prostatectomy and bilateral inguinal lymph node dissection was reported to be pT3N0 and whole body scanning for metastases, was negative. PMID- 18386165 TI - SDF-1 and CCR5 genes polymorphism in patients with head and neck cancer. AB - The frequency of SDF1-3'A and CCR5Delta32 in patients with head and neck cancer were determined in this study. The frequencies of alleles and genotypes of SDF-1 and CCR5 were assessed by PCR method in 156 patients with malignant head and neck cancer, 125 (80.1%) cases with squamous cell carcinoma and 31 (19.9%) cases with salivary gland tumors and compared with 262 age-sex matched healthy control individuals. SDF-1 genotypes in patients with SCC of head and neck, but not with salivary gland tumors, showed a statistically significant difference compared to the normal group (P < 0.005 for SCC and P = 0.3 for salivary gland tumors). There were no significant differences in the frequencies of SDF1-3'A allele, CCR5 genotypes and alleles between patients and controls. Based on the present study SDF1-3A may be associated with the susceptibility of patients to SCC of head and neck cancer. PMID- 18386166 TI - Expression of matrilin-2 in liver cirrhosis and hepatocellular carcinoma. AB - The recently described matrilin protein family is part of the extracellular matrix, their pathophysiological role as well as distribution in liver diseases, however, have not yet been studied. Considering that matrilins have been found to play role in cell growth and tissue remodeling, their possible involvement in carcinogenesis has been raised. The main objective of this study was to investigate the changes in matrilin-2 expression which is one of the main components of basement membranes. Thirty-five cases of surgically resected hepatocellular carcinomas, 35 corresponding surrounding liver tissues and 10 normal liver samples were used for the study. In 15 of 35 cases the tumor developed on the basis of cirrhosis. Matrilin-2 protein expression was detected in normal liver around bile ducts, portal blood vessels, while sinusoids were negative by immunohistochemistry. Cirrhotic surrounding tissue showed intensive matrilin-2 staining along the sinusoids. Tumorous neovasculature was found strongly positive by immunohistochemistry. No differences, however, were detected by morphometry regarding the amount of protein expression based on the grade of hepatocellular carcinomas. Real-time RT-PCR did not show significant differences in matrilin-2 mRNA expression between normal, cirrhotic and tumor samples. This suggests posttranslational modification of matrilin-2 manifesting in altered distribution in liver fibrosis. Our data indicate that matrilin-2 is a novel basement membrane component in the liver, which is synthetised during sinusoidal "capillarization" in cirrhosis and in hepatocellular carcinoma. This is the first report to describe the expression and distribution of matrilin-2 in human normal and cirrhotic liver as well as in hepatocellular carcinoma. PMID- 18386168 TI - Differentiating subgroups of children with special health care needs by health status and complexity of health care needs. AB - OBJECTIVES: Our objective is to use the Children with Special Health Care Needs (CSHCN) Screener to identify subgroups of CSHCN differentiated by health status and complexity of need. METHODS: Data are from the National Survey of Children with Special Health Care Needs, 2001 and the National Survey of Children's Health, 2003 (conducted by the Maternal and Child Health Bureau and the National Center for Health Statistics); and the 2001 and 2002 Medical Expenditure Panel Survey, conducted by the Agency for Healthcare Research and Quality. A broad array of variables measuring health status, complexity of need, and related issues are examined by subgroupings of CSHCN. RESULTS: Relative to other CSHCN, CSHCN with functional limitations or who qualify on more CSHCN Screener items have poorer health status and more complex health care needs. They more often experience a variety of health issues; their insurance is more often inadequate; the impact of their conditions on their families is higher; and their medical costs are higher. CONCLUSION: In the absence of information on specific conditions, health status, or complexity of need, the CSHCN Screener alone can be used to create useful analytic subgroups that differ on these dimensions. The proposed subgroups, based on the type or number of CSHCN screening criteria, differentiate CSHCN by health status and complexity of health care needs, and also show differences in the impact of their conditions on their families, costs of their medical care, and prevalence of various health problems. PMID- 18386169 TI - Prospective comparison of the diagnostic utility of a standard event monitor versus a "leadless" portable ECG monitor in the evaluation of patients with palpitations. AB - INTRODUCTION: Current ambulatory ECG monitoring systems are limited in their ability to diagnose patients with palpitations. The aim of this prospective study was to compare a new "leadless" ambulatory monitor with a standard event monitor in the evaluation of patients with palpitations. METHODS: Eighteen consecutive patients (11 female, 56 +/- 16 years) referred for evaluation of palpitations were provided with both a standard event monitor and a "leadless" monitor for 30 days. They were asked to record episodes of palpitations with both monitoring devices. RESULTS: All 18 individuals were compliant with the "leadless" monitor for the 30-day period while only 14 (78%) patients were compliant with the standard event monitor (p = 0.10). During a combined monitoring period of 563 days, 159 symptomatic episodes were recorded with the "leadless" ECG monitor (8.8 +/- 9.7 per patient, range 1-35) and 169 symptomatic episodes were recorded with the event monitor (12 +/- 8.3 per patient, range 1-33) (p = NS). The "leadless" ECG monitor recorded arrhythmias in 13 of 18 patients (72%) and the standard event monitor recorded arrhythmias in 8 of 14 patients (57%) (p = NS). CONCLUSION: The "leadless" ECG monitor is associated with high patient compliance and results in high quality ECG recordings. The diagnostic yield of this monitoring system is equivalent to a standard event monitor. PMID- 18386170 TI - Advanced mapping techniques in atrial fibrillation. AB - Catheter ablation has emerged as an excellent treatment option for atrial fibrillation especially in patients with paroxysmal AF. Several obstacles however remain regarding ablation strategies for persistent and chronic AF. In this setting, adequate success rates can only be achieved with left atrial ablation in addition to complete PV isolation. Important techniques in this regard are mapping of complex fractionated atrial electrograms as well as identification of atrial sites serving as sources of persistent AF. Other mapping techniques reviewed in this article are rapid geometry acquisition with spiral catheters and the Ensite/NAVX system, the 64-pole Constellation basket catheter as well as the MESH Mapper catheter. PMID- 18386171 TI - Aging and social change among Abaluyia in western Kenya: anthropological and historical perspectives. AB - This article presents narrative accounts which illustrate ways that modernization and social change have transformed the daily lives of Abaluyia, especially older people, in rural western Kenya since the late nineteenth century. The narratives reveal history as lived experience, as observed and recorded by an anthropologist who has been doing research among Abaluyia in Bunyala and Samia over the past 25 years. The story involves continuity of cultural beliefs and practices, and it involves change-change imposed by the macro-events of globalizing processes, from colonialism to Structural Adjustment Programs, and change as people's adaptive responses to those processes, particularly how changing cultural practices have impacted elders. The grand narrative is historical, the overarching story of the incorporation of Kenya and Kenyans into the global political economy from the colonial period to the present. Other narratives are biographical, case studies of individuals from two extended families and their personal experiences of social change over the past century. The background narrative is autobiographical, the anthropologist's story of the practice of anthropological fieldwork and her own experiences and observations of social change in western Kenya. Since anthropological data over time become history, the approach here is both anthropological and historical. PMID- 18386172 TI - Investigation of tRNA(Leu/Lys) and ATPase 6 genes mutations in Huntington's disease. AB - Huntington disease (HD) is a genetically dominant condition caused by expanded CAG repeats which code for glutamine in the HD gene product, huntingtin. Huntingtin is expressed in almost all tissues, so abnormalities outside the brain can also be expected. Involvement of nuclei and mitochondria in HD pathophysiology has been suggested. In fact mitochondrial dysfunction is reported in brains of patients suffering from HD. The tRNA gene mutations are one of hot spots that can cause mitochondrial disorders. In this study, possible mitochondrial DNA (mtDNA) damage was evaluated by screening for mutations in the tRNA(leu/lys) and ATPase 6 genes of 20 patients with HD, using PCR and automated DNA sequencing. Mutations including an A8656G mutation in one patient were observed, which may be causal to the disease. Understanding the role of mitochondria in the pathogenesis of neurodegenerative diseases could potentially be important for the development of therapeutic strategies in HD. PMID- 18386174 TI - The surgical margin status after breast-conserving surgery: discussion of an open issue. AB - Hypothesis The best therapeutic approach to the involved or proximal surgical margins has not been defined yet; surgical margins status can influence the local relapse of disease in breast carcinoma, but the impact on overall survival has not been clearly demonstrated. Purpose of this work is to find in the available literature further evidence to guide the therapeutic behaviour in patients with close margins by invasive carcinoma. Design Review of the currently available literature on the evaluation of surgical margins in breast conserving surgery; influence of margin involvement by invasive component or intraductal component. Patients or other participants Literature research by PubMed on the topics of breast carcinoma, conservative surgery and margin definition and status; therapeutic approach to involved margins. Main outcome measure We reviewed the available literature focusing our attention to the definition of clear surgical margins and to the value of the close proximity of margins in relation to the local control of disease and the best therapeutic management of different situations. Results Further evidence is needed on large numbers of patients to understand how to evaluate surgical margins in invasive breast carcinoma. Conclusions There is no consensus on the definition of "clear surgical margins", and the ideal approach to the close proximity of margins has not been defined. It is not sure whether a new surgical procedure is really needed in every case of close proximity of tumor cells to the margins. Radiation therapy could be a good option in the management of these cases, but further evidence is needed to establish the real impact of clear surgical margins on local control of disease and, furthermore, on survival. PMID- 18386173 TI - Programmed cell death 4 inhibits breast cancer cell invasion by increasing tissue inhibitor of metalloproteinases-2 expression. AB - High levels of the cyclooxygenase-2 (COX-2) protein have been associated with invasion and metastasis of breast tumors. Both prostaglandin E(2) (PGE(2)) and interleukin-8 (IL-8) have been shown to mediate the invasive activity of COX-2 in breast cancer cells. Here we expand these studies to determine how COX-2 uses PGE(2) and IL-8 to induce breast cancer cell invasion. We demonstrated that PGE(2) and IL-8 decreased the expression of the tumor suppressor protein Programmed Cell Death 4 (PDCD4). We hypothesized that suppression of PDCD4 expression is vital to the invasive activity of PGE(2) and IL-8. In MCF-7 cells overexpressing PDCD4 (MCF-7/PDCD4), PGE(2) and IL-8 failed to induce invasion, in contrast to the parental MCF-7 cells, thus indicating that PDCD4 blocks breast cancer cell invasion. MCF-7/PDCD4 cells produced higher levels of the Tissue Inhibitor of Metalloproteinases-2 (TIMP-2) than the parental cells. Silencing TIMP-2 mRNA in MCF-7/PDCD4 cells reversed the anti-invasive effects of PDCD4, allowing PGE(2) and IL-8 to induce the invasion of these cells. Here we report the novel findings that suppression of PDCD4 expression is vital for the invasive activity of COX-2 mediated by PGE(2) and IL-8, and that PDCD4 increases TIMP-2 expression to inhibit breast cancer cell invasion. PMID- 18386175 TI - Variation of delta13C in Aegiceras corniculatum seedling induced by cadmium application. AB - To test whether the values of delta13C in mangrove plants are affected by Cd application, the seedlings of Aegiceras corniculatum, a dominant mangrove species, were cultured in soil supplied with CdCl2 solution at the concentration of 0, 0.5, 2.5, 5, 20, 30 and 50 CdCl2 mg/kg wet soils. Plants were grown in 10 replicate pots with 5 propagules each. After 5 months of Cd exposure, three pots contained 15 seedlings with average shoot heights were selected for each treatment. Leaves and roots of seedling were sampled respectively and analyzed for delta(13)C. Growth traits (fresh weight, shoot height and root length), total chlorophyll content, and Cd concentrations in leaf, root and dry soil were determined. After 5 months of the seedling growth, the concentrations of Cd in dry soil were 0.47, 0.83, 2.77, 4.54, 18.89, 29.79 and 47.35 mg/kg respectively. The values of delta13C in roots and leaves were affected to some extent by Cd application. Although root delta(13)C showed more sensitive to Cd compared with leaves, the values of delta13C in roots were not significantly affected by Cd until Cd level higher than 29.78 mg/kg which was not expected to exist in natural environments. Minor variation in delta13C values observed in roots and leaves was likely due to limited Cd uptake by seedlings and subsequent lack of negative impacts on photosynthesis. PMID- 18386176 TI - Molecular targets of anxiety: from membrane to nucleus. AB - Anxiety is a common human emotional experience that causes decreased quality of life and increased social burden worldwide. However, the treatment options currently available for anxiety are limited as the molecular mechanisms of these complicated emotional disorders are poorly understood. With the development of integrative methods including genetic manipulations, a variety of molecular targets involved in anxiety have been revealed, from membrane receptors, such as 5-HT receptor, GABA(A) receptor and GluR5 kainate receptor, and intracellular signaling proteins, such as CaMKIV and AC8, to transcription factors, such as CREB and Egr-1. We propose that all these molecules act together to form a balance between excitatory and inhibitory transmission that is critical for physiological anxiety, and that prolonged disturbance of any of them can promote pathological anxiety-like behavior. Studies on the interactions between these molecules will help elucidate the cellular mechanisms of anxiety, and will provide molecular targets for treating the disorders. PMID- 18386177 TI - Assessing child and adolescent pragmatic language competencies: toward evidence based assessments. AB - Using language appropriately and effectively in social contexts requires pragmatic language competencies (PLCs). Increasingly, deficits in PLCs are linked to child and adolescent disorders, including autism spectrum, externalizing, and internalizing disorders. As the role of PLCs expands in diagnosis and treatment of developmental psychopathology, psychologists and educators will need to appraise and select clinical and research PLC instruments for use in assessments and/or studies. To assist in this appraisal, 24 PLC instruments, containing 1,082 items, are assessed by addressing four questions: (1) Can PLC domains targeted by assessment items be reliably identified?, (2) What are the core PLC domains that emerge across the 24 instruments?, (3) Do PLC questionnaires and tests assess similar PLC domains?, and (4) Do the instruments achieve content, structural, diagnostic, and ecological validity? Results indicate that test and questionnaire items can be reliably categorized into PLC domains, that PLC domains featured in questionnaires and tests significantly differ, and that PLC instruments need empirical confirmation of their dimensional structure, content validity across all developmental age bands, and ecological validity. Progress in building a better evidence base for PLC assessments should be a priority in future research. PMID- 18386178 TI - Determinants of mental health consultations among recent Chinese immigrants in British Columbia, Canada: implications for mental health risk and access to services. AB - Linked immigration and health administrative databases were analyzed to study the factors associated with the rate of mental health consultations with physicians in 1992-2001 of over 150,000 Chinese immigrants in British Columbia, Canada. Results showed that number of years since landing and rate of non-mental health visits to general practitioners were the most consistent variables associated with mental health consultations to general practitioners and psychiatrists in all sex and age groups. Other variables associated with the rate of consultations were age, place of origin, educational level, marital status and English skill. Supply of physicians was not observed to be associated with mental health consultations. The findings are consistent with Andersen's behavioral model of health care utilization and introduce components specially pertinent to immigrants and mental health service utilization. They also highlight sub populations among immigrants who may be at risk of experiencing mental health problems or encountering barriers to care. PMID- 18386179 TI - The use of complementary and alternative medicine among Chinese Canadians: results from a national survey. AB - This study examines the relationship between Chinese Canadian ethnicity and the use of complementary and alternative medicine (CAM) and explores some of the factors that contribute to CAM use among this visible minority group. Using data from cycle 2.1 of the Canadian Community Health Survey (2003), we use multivariate logistic regression to investigate the extent to which CAM use varies among Chinese Canadians and non-Chinese Canadians. Two three-way interactions, which demonstrate how the combination of certain identity markers increases their predictive value within the model, are also examined. The use of CAM varies according to ethnicity, with Chinese Canadians being more likely to use than non-Chinese Canadians. The findings also indicate that cultural factors play a key role in establishing the necessary conditions for increasing the likelihood of CAM use for Chinese Canadians. Findings are discussed in terms of their implications for health care policy and program development. PMID- 18386181 TI - Theory of planned behavior and multivitamin supplement use in Caucasian college females. AB - The objective of this study was to identify predictors of the use of multivitamin supplements (MVS) among Caucasian college females utilizing the Theory of Planned Behavior (TPB). Variables of the TPB and the self-reported use of multivitamin supplements were measured by two separate surveys within 1 week with a convenience sample of 96 Caucasian college student females. Two attitudinal beliefs and one control belief significantly predicted behavioral intention to use multivitamin. A belief that taking multivitamin supplements helps to feel and look good was the most important predictor of the use of multivitamin supplements. EDITORS' STRATEGIC IMPLICATIONS: Findings from this study, although in need of replication, suggest that prevention campaigns would be more successful if messages used to reach these females were consistent with perceived beliefs regarding benefits of using MVS. More broadly, TPB appears to offer a useful framework for understanding or predicting behavior based on psychological constructs theorized to influence behavior. PMID- 18386180 TI - Correlates and predictors of tobacco use among immigrant and refugee youth in a Western Canadian city. AB - BACKGROUND: Tobacco control is a priority of the British Columbia Ministry of Health as illnesses associated with tobacco use are the leading cause of preventable death in the province. As a result of increased immigration, British Columbia's demographic profile is becoming more diverse and necessitates approaches to health promotion and disease prevention that are culturally relevant. In order to develop culturally relevant anti-smoking messages and resources for immigrant and refugee youth, surveys were administered to 194 youth to better understand their attitudes towards smoking and to explore predictors of tobacco use. RESULTS: Twelve percent of respondents reported smoking all or part of a cigarette within the past 30 days. Male respondents were three times more likely to smoke than female respondents. Logistic regression analysis showed that immigrant and refugee youth were more likely to be non-smokers if they did not have a father who smokes, drank alcohol less frequently and had fewer close friends who smoke. IMPLICATIONS: These findings support previous research studies that relate youth smoking to social influences and demonstrate a need to address gender differences, the confluence of smoking and drinking and the significance of family and peer pressure on smoking when designing culturally relevant anti smoking resources. PMID- 18386182 TI - The tubulin-depolymerising agent combretastatin-4 induces ectopic aster assembly and mitotic catastrophe in lung cancer cells H460. AB - The relationship between microtubular dynamics, dismantling of pericentriolar components and induction of apoptosis was analysed after exposure of H460 non small lung cancer cells to anti-mitotic drugs. The microtubule destabilising agent, combretastatin-A4 (CA-4) led to microtubular array disorganization, arrest in mitosis and abnormal metaphases, accompanied by the presence of numerous centrosome-independent "star-like" structures containing tubulin and aggregates of pericentrosomal matrix components like gamma-tubulin, pericentrin and ninein, whereas the structural integrity of centrioles was not affected by treatment. On the contrary, in condition of prolonged exposure or high concentrations of CA-4 such aggregates never formed. Treatment with 7.5 nM CA-4, which produced a high frequency "star-like" aggregates, was accompanied by mitotic catastrophe commitment characterized by translocation of the proapoptotic Bim protein to mitochondria activation of caspases-3/9 and DNA fragmentation as a result of either prolonged metaphase arrest or attempt of cells to divide. Drug concentrations which fail to block cells at mitosis were also unable to activate apotosis. A detailed time-course analysis of cell cycle arrest and apoptosis indicated that after CA-4 washout the number of metaphases with "star-like" structures decreased as a function of time and arrested cells proceeded in anaphase. After 4 h, the multiple alpha- and gamma-tubulin aggregates coalesced into two well-defined spindles in a bipolar mitotic spindle organization. Overall, our findings suggest that the maintenance of microtubular integrity plays a relevant role in stabilising the pericentriolar matrix, whose dismantling can be associated with apoptosis after exposure to microtubule depolymerising agents. PMID- 18386183 TI - Rhizobacteria in mycorrhizosphere improved plant health and yield of banana by offering proper nourishment and protection against diseases. AB - The corporate R&D banana orchards of Musa paradisiaca (dwarf Cavendish AAA, var. shrimanti) on a medium black alluvial soil with low nutrients harboured diversified species of vesicular-arbuscular mycorrhizal (VAM) fungi. These fungi infected the roots severely (69.2%), showed elevated (69.8 g(-1) soil) spore density, increased soil bacterial density (245 x 10(8) cfu g(-1)), produced siderophores (58.2%) and reduced nematode population (2.3 g(-1)) in the mycorrhizosphere of plants for integrated plant nutrition management (IPNM) system as compared to traditional treatment of applying chemical fertilisers alone and other test treatments. The interactions of plant roots with native VAM and local and applied rhizobacteria in the matrix of soil conditioner enabled proper nourishment and protection of crop in IPNM treatment as compared to traditional way. Hence, exploitation of plant growth promoting rhizobacteria through judiciously designed IPNM system revealed the (a) relatively increased banana productivity (21.6%, 76 MT ha(-1)), (b) least occurrence of fusarial wilt and negligible evidence of Sigatoka, (c) saving of 50% chemical fertilisers and (d) permitted control over soil fertility in producer's favour over traditional cultivation practices. These findings are discussed in detail. PMID- 18386184 TI - Batch fermentation model of propionic acid production by Propionibacterium acidipropionici in different carbon sources. AB - Propionic acid (PA) is widely used as additive in animal feed and also in the manufacturing of cellulose-based plastics, herbicides, and perfumes. Salts of propionic acid are used as preservative in food. PA is mainly produced by chemical synthesis. Nowadays, PA production by fermentation of low-cost industrial wastes or renewable sources has been an interesting alternative. In the present investigation, PA production by Propionibacterium acidipropionici ATCC 4965 was studied using a basal medium with sugarcane molasses (BMSM), glycerol or lactate (BML) in small batch fermentation at 30 and 36 degrees C. Bacterial growth was carried out under low dissolved oxygen concentration and without pH control. Results indicated that P. acidipropionici produced more biomass in BMSM than in other media at 30 degrees C (7.55 g l(-1)) as well as at 36 degrees C (3.71 g l(-1)). PA and biomass production were higher at 30 degrees C than at 36 degrees C in all cases studied. The best productivity was obtained by using BML (0.113 g l(-1) h(-1)), although the yielding of this metabolite was higher when using glycerol as carbon source (0.724 g g(-1)) because there was no detection of acetic acid. By the way, when using the other two carbon sources, acetic acid emerged as an undesirable by-product for further PA purification. PMID- 18386185 TI - Oligodendrocytes take center stage. Introduction. PMID- 18386186 TI - Expression of gLTP in sympathetic ganglia from stress-hypertensive rats: molecular evidence. AB - We previously reported behavioral and electrophysiological evidence indicating that superior cervical ganglia (SCG) from rats that developed hypertension as a result of chronic psychosocial stress expressed ganglionic long-term potentiation (gLTP) in vivo. In the present study, we present additional supportive evidence by measuring changes in protein levels of essential signaling molecules in ganglia from chronically stressed rats. We compared protein levels of essential, LTP-related signaling molecules in ganglia isolated from chronic stress hypertensive rats, known to have expressed gLTP, with those of the same molecules in normal ganglia 1h after eliciting gLTP by high frequency stimulation (HFS) in vitro. Immunoblot analysis showed a significant increase in the levels of phosphorylated CaMKII, total CaMKII, nitric oxide synthase (NOS-1), and calmodulin in SCG from both chronically stressed rats and from normal rat ganglia in which gLTP was expressed by HFS in vitro. Additionally, there was a parallel reduction in calcineurin protein levels in ganglia from both groups. The present results confirm that ganglia from stressed rats have expressed gLTP in vivo and that synaptic plasticity in sympathetic ganglia may involve a molecular cascade largely similar to that of LTP in the hippocampal CA1 region. PMID- 18386187 TI - Antifibrinolytic therapy to prevent early rebleeding after subarachnoid hemorrhage. AB - In the current era of early surgery, there has been little interest in the use of antifibrinolytic therapy to prevent rebleeding after aneurysmal subarachnoid hemorrhage (aSAH). Older studies demonstrated that antifibrinolytics can reduce rebleeding, but long-term therapy results in increased cerebral ischemia from vasospasm, leading to no appreciable effect on mortality. While early surgery would seem to obviate the need for long-term antifibrinolytic use, a subgroup of patients may benefit from early therapy. The rate of pre-operative rebleeding may be as high as 9-17%, causing significant morbidity and mortality. Short-term use of antifibrinolytic agents (less than 3 days) in the presence of calcium channel blocking therapy has shown promising results. A randomized clinical trial of early antifibrinolytic therapy conducted in 2002 showed a significant decrease in rebleeding and a non-significant decrease in overall mortality. In this review, we examine the clinical pharmacology, dosing, monitoring, complications, and side effects of antifibrinolytic treatment. We conclude that early short-term antifibrinolytic therapy might be a reasonable strategy to prevent acute rebleeding and improve long-term outcome in aSAH patients. Additional randomized clinical trials are necessary to determine whether this management strategy is effective. PMID- 18386188 TI - Gene expression profiles of mouse striatum in control and maneb + paraquat induced Parkinson's disease phenotype: validation of differentially expressed energy metabolizing transcripts. AB - The present study was undertaken to investigate the gene expression patterns of the striatum of control and maneb + paraquat-induced Parkinson's disease (PD) phenotype in mouse to identify the differentially expressed transcripts. The animals were treated with and without maneb (30 mg/kg, i.p.) + paraquat (10 mg/kg, i.p.), twice a week, for 3, 6, and 9 weeks. The RNA was isolated from control and treated mouse striatum and reverse transcribed, and equal quantities of labeled cDNA were mixed and hybridized with mouse 15 k arrays. Comparative transcription patterns showed the time of exposure dependent alteration in the expression of several transcripts associated with various pathways. RT-PCR reconfirmed the differential expression of some energy metabolizing transcripts. The study provides maneb + paraquat-induced differential expression of many transcripts using high-density microarray approach. Few transcripts, which were previously not reported to be associated with neuronal degeneration, were also identified. The results obtained thus suggest that maneb + paraquat induce neurotoxicity in the striatum in a time of exposure dependent manner via multiple pathways and defective energy metabolism could play a critical role. PMID- 18386189 TI - Effect of background colour on growth and adhesion strength of Ulva sporelings. AB - This study examined the effects of a range of black, grey and white substrata on the growth and attachment strength of Ulva sporelings on glass and polydimethylsiloxane (Silastic-T2) surfaces. The rate of development of sporelings was strongly influenced by the colour of the substratum on which they grew. On black backgrounds, sporelings grew slowly and germination was delayed. Laboratory screening methods for antifouling and fouling-release coatings that rely on the growth of Ulva sporelings can be compromised if samples are of different colours. Hydrodynamic removal of sporelings from coatings may also be affected by substratum colour, since smaller plants generate lower hydrodynamic forces making them more difficult to remove. PMID- 18386190 TI - A case of impaired verbalization but preserved gesticulation of motion events. AB - In most cultures, most of the time, when people talk they gesture. We took advantage of a rare opportunity to explore the relation between the verbalization and gesticulation of motion events by studying Marcel, an English speaker with a unilateral left-hemisphere lesion affecting frontal, parietal, and temporal sectors of the perisylvian cortex. Marcel has intact semantic knowledge of the three major classes of words that are commonly used in English descriptions of motion events - specifically, concrete nouns, action verbs, and spatial prepositions - as well as intact syntactic knowledge of how these word classes are typically combined in the intransitive motion construction (e.g., The ball rolled down the hill). However, his ability to retrieve the lexical-phonological structures of these words is severely impaired. Despite this profound anomia, he is still remarkably skilled at producing iconic manual depictions of motion events, as demonstrated in two experiments involving spontaneous gestures and one experiment involving elicited gestures. Moreover, the structural characteristics of Marcel's gestures are clearly sensitive to the idiosyncratic meanings of English verbs and prepositions, and they may also be sensitive to the way motion events are syntactically packaged in the intransitive motion construction. These findings improve our understanding of how some brain-damaged individuals with severe aphasia but without manual apraxia can successfully employ gesture to augment the semantic content of their speech. PMID- 18386191 TI - A quantitative overview of radiosensitivity of human tumor cells across histological type and TP53 status. AB - PURPOSE: We have previously shown in a limited number of tumor cell lines derived from only two histological types that clonogenic survival patterns fall into radiosensitivity groups, each group associating with a specific genotype. We now establish a global, quantitative description of human tumor cells based on genotype-dependent radiosensitivity across histological types. METHODS: We measure clonogenic radiosensitivity in 39 human tumor cell lines that vary in histological type (colorectal, glioblastoma, prostate, bladder, teratoma, breast, melanoma and liver) and expression of several genes purported to influence radiosensitivity: ATM (ataxia telangiectasia mutated), TP53 (tumor protein 53), CDKN1A (cyclin-dependent kinase N1A), 14-3-3sigma (an isoform of the 14-3-3 gene) and DNA mismatch repair genes . For each survival curve we use the linear quadratic model and a linear-linear model to extract multiple coefficients and seek correlation across histological types. RESULTS: Under one-parameter analysis, survival rate at circa 2 Gy, cell lines segregate into two major, statistically-significant groups that correlate with TP53 status (wildtype versus mutant). Under two-parameter analysis, cell lines segregate into four radiosensitivity groups based on correlations between response at lower doses (ca. 2 Gy) and a component of response to higher doses (>4 Gy). CONCLUSIONS: Intrinsic radiosensitivity of 39 human tumor cell lines segregate into distinct genotype-dependent radiosensitivity groups that associate with mutATM, wtTP53, mutTP53, and an unidentified factor in some glioblastoma cells. Genotype dependent radiosensitivity underlies histology-dependent variation in radiosensitivity. Our analysis establishes a quantitative overview of radiosensitivity that can predict possible response of human tumors to radiotherapy protocols. PMID- 18386192 TI - Radioresistance in a tumour cell line correlates with radiation inducible Ku 70/80 end-binding activity. AB - PURPOSE: The aims of the present study were to better understand the role of Ku 80, which is involved in double-strand break repair in mammalian cells in the mechanism of radiation resistance and to verify the possibility of increasing cell radiosensitivity by targeted inhibition of Ku autoantigen 80 (Ku 80). MATERIALS AND METHODS: Western blot and electrophoretic mobility shift assay (EMSA) were performed on the human bladder carcinoma cell line RT112 (radioresistant) and on the human colorectal carcinoma cell line SW48 (radiosensitive) to assess the expression levels of DNA-dependent protein kinase (DNA-PK) components and the DNA-binding activity of the Ku 70/80 heterodimer after exposure to radiation, respectively. Ku 80 silencing was carried out with the use of small interfering RNA (siRNA). RESULTS: Greater differences in the DNA binding activity of Ku 70/80 and Ku 80 phosphorylation level were observed in RT112 as compared to SW48 after X-ray treatment. There is no correlation between Ku expression and DNA-binding activity at lower doses. A significant increase in nuclear Ku 80 expression was observed one hour after the exposure, only at the higher doses, while the DNA-PK catalytic subunits (DNA-PKcs) and Ku 70 levels did not change significantly. Inhibition of Ku 80 expression by siRNA induced radiosensitivity in the RT112 cell line. CONCLUSIONS: Our data demonstrate that in a bladder tumour cell line up-regulation of Ku end-binding activity without any marked change in Ku expression underlie radiation resistance. PMID- 18386193 TI - Lymphocyte telomere length correlates with in vitro radiosensitivity in breast cancer cases but is not predictive of acute normal tissue reactions to radiotherapy. AB - PURPOSE: To examine the hypothesis that lymphocyte telomere length may be predictive of both breast cancer susceptibility and severity of acute reactions to radiotherapy. MATERIALS AND METHODS: Peripheral blood lymphocyte cultures from breast cancer patients (with normal or severe skin reactions to radiotherapy) and normal individuals were assessed for in vitro radiosensitivity as measured by apoptosis, cell cycle delay and cytotoxicity. Telomere lengths were determined by a flow cytometric fluorescence in situ hybridization assay (FLOW-FISH). RESULTS: Female breast cancer cases (n = 24) had reduced lymphocyte telomere lengths by comparison with healthy controls (n = 20, p < 0.04). However, the average age of healthy controls was less (45.4) than cases (53). When the control group was modified to give a better age match (51.5, n = 13) the reduced telomere length in cases was not significantly different from controls. Lymphocytes from breast cancer cases also showed reduced cell cycle delay (p < 0.001) and increased apoptosis (p < 0.01) following irradiation in vitro at 3 and 5 Gy respectively, compared to healthy controls. Statistical significance was maintained with the improved age matching of groups. Comparison of lymphocytes from breast cancer patients with normal (n = 11) and severe (n = 13) skin reactions to radiotherapy failed to identify differences in telomere length or cellular radiosensitivity in this limited sample. CONCLUSIONS: This study adds to the evidence suggesting a correlation between altered cellular radiosensitivity and breast cancer. However, in the cases investigated, telomere length does not appear to be predictive of acute skin reactions to radiotherapy. PMID- 18386194 TI - Irradiation as preparative regimen for hepatocyte transplantation causes prolonged cell cycle block. AB - PURPOSE: Hepatocyte transplantation following liver irradiation (IR) and partial hepatectomy (PH) leads to extensive liver repopulation. We investigated the changes in the liver induced by IR explaining the loss of reproductive integrity in endogenous hepatocytes. MATERIALS AND METHODS: Right lobules of rat liver underwent external beam IR (25 Gy). A second group was subjected to additional 33% PH of the untreated left liver lobule. Liver specimens and controls were analyzed for DNA damage, apoptosis, proliferation and cell cycle related genes (1 hour to up to 12 weeks). RESULTS: Double strand breaks (phosphorylated histone H2AX) induced by IR rapidly declined within hours and were no longer detectable after 4 days. No significant apoptosis was noted and steady mRNA levels (B-cell lymphoma 2-associated X protein (BAX), caspase 3 and 9) were in line with the lack of DNA fragmentation. However, gene expression of p53 and p21 in irradiated liver tissue increased. Transcripts of cyclin D1, proliferating cell nuclear antigen (PCNA), and cyclin B augmented progressively, whereas cyclin E was only affected moderately. Following PH, irradiated livers displayed persistently high protein levels of p21 and cyclin D1. However, cell divisions were infrequent, as reflected by low PCNA levels up to four weeks. CONCLUSION: IR leads to a major arrest in the G(1)/S phase and to a lesser extent in the G(2)/M transition of the cell cycle, resulting in reduced regenerative response following PH. The persistent block of at least four weeks may promote preferential proliferation of transplanted hepatocytes in this milieu. PMID- 18386195 TI - Gene expression analysis in human malignant melanoma cell lines exposed to carbon beams. AB - PURPOSE: To elucidate the molecular changes in response to carbon beams (C-ions) in melanoma. MATERIALS AND METHODS: We examined expression profiles of 6 melanoma cell lines exposed to C-ions or X-rays with 2 Gy using single-color microarrays. RESULTS: Twenty-two genes, including nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (NFKBIA), responded to C-ions in all six cell lines, based on analysis of variance (ANOVA) filtering (p < 0.001). We found 173 genes that responded in common to C-ions in four cell lines. We identified many down-regulated genes including the cell cycle - related genes that were more responsive to C-ions than X-rays. In contrast, most of the up regulated genes including the tumor protein p53 (p53) target genes responded to both C-ions and X-rays. C-ions induced G2/M arrest significantly more than X-rays at 30 h (p < 0.05). CONCLUSION: Our findings suggest that down-regulation of gene expression plays a key role in the response to C-ions. Regulation of cell cycle - related genes and induction of prolonged G2/M arrest may be responsible for the extra sensitivity to C-ions, whereas p53-related genes may have similar roles in the sensitivities to both C-ions and X-rays. PMID- 18386196 TI - Expression of activated ATF-2, CREB and c-Myc in rat colon transversum after whole-body gamma-irradiation and its contribution to pathogenesis and biodosimetry. AB - PURPOSE: The purpose of our study is to examine phospho-ATF-2(Thr-69/71) (phospho activating transcription factor-2, p-ATF-2), phospho-CREB(Ser-133) (phospho-cAMP response binding element protein, p-CREB), and phospho-c-Myc(Thr-58/Ser-62) (phosho-myelocytomatosis protooncogene, p-c-Myc) expression in irradiated rat colon transversum. MATERIALS AND METHODS: Male Wistar rats were randomly divided to 28 groups and irradiated with whole-body gamma-radiation of 0, 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 Gy. Samples were taken 4 and 24 hours after the irradiation, immunohistochemically stained. P-ATF-2, p-CREB, and p-c Myc expression was measured. RESULTS: We measured increased cytoplasmatic p-ATF-2 expression 4 hours after irradiation by 0.25 - 1, 10 Gy and 24 hours after irradiation by 0.5 - 1, 10 Gy. Increased cytoplasmatic p-CREB expression was found 4 hours after irradiation by 0.25 - 1, 9, 10 Gy and 24 hours after irradiation by 0.25 - 1, 4, 10 Gy. Increased p-c-Myc cytoplasmatic expression was found 4 hours after irradiation by 0.25, 0.75, 4, 5 Gy and 24 hours after irradiation by 0.75, 1, 10 Gy. Nuclear p-ATF-2, p-CREB, and p-c-Myc expressions were similar to their cytoplasmatic expressions. CONCLUSION: The detection of p ATF-2 and p-CREB might be considered as a perspective biodosimetric tool for irradiated enterocytes in vivo. The use of p-c-Myc appears to be controversial due to the ambivalent expression values. PMID- 18386197 TI - Altered blood chemistry and hippocampal histomorphology in adult rats following prenatal exposure to physiologically-patterned, weak (50-500 nanoTesla range) magnetic fields. AB - PURPOSE: To discern changes in blood chemistry, cerebral sizes, and hippocampal cytomorphology in adult male and female albino Wistar rats that had been exposed during their entire prenatal development to one of two patterns of magnetic fields and one of four intensities: Very low 5 - 20 nT; low 30 - 50 nT; medium 90 - 580 nT; and high 590 nT to 1.2 microT. MATERIALS AND METHODS: A total of 48 pregnant females were exposed to either a repetitive frequency-modulated magnetic field or to a complex sequence of 50, 200-msec physiologically-patterned fields. As adults blood, cerebral, and histomorphological data were obtained from the 137 rats that had been exposed to one of these eight conditions. RESULTS: Compared to other groups, adult rats that had been exposed prenatally to the physiologically patterned magnetic fields at the low (30 - 50 nT) and medium (90 - 580 nT) intensities exhibited peak elevations of aminotransaminase, glucose, and uric acid. Numbers of cytometric anomalies were also significantly elevated within regions of the hippocampus known for neuronal neogenesis in adults. CONCLUSIONS: The results suggest that a common factor in cellular adhesion or plasticity might be permanently altered by prenatal exposure to a narrow intensity of a series of physiologically-patterned magnetic fields. PMID- 18386198 TI - Thorium-induced oxidative stress mediated toxicity in mice and its abrogation by diethylenetriamine pentaacetate. AB - PURPOSE: Thorium ((232)Th, IV) preferentially accumulates in the liver, femur and spleen, which necessitates evaluation of its toxic effect in these organs. The present study was aimed at evaluation of liver function, oxidative stress and histological alterations in these organs. MATERIALS AND METHODS: Swiss albino mice were administered either with Thorium nitrate (10 mg/kg body weight/day equivalent to 1,090 pCi/kg body weight/day) for 30 days (1/40th dose of LD(50/30); the dose of thorium required to kill 50% of the test cohort within 30 days) intraperitoneally or with calcium salt of diethylenetriamine pentaacetate (Ca-DTPA, 100 micromole/kg body/weight) intravenously or both. Liver function tests and oxidative damage was assessed. The concentration of Th in the tissues was determined by Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP AES) method. RESULTS: Administration of Th prevented the increase in the body and liver weight and altered liver functions. Th treatment to mice showed a decrease in the activities and gene expression of antioxidant enzymes, and increased lipid peroxidation and protein carbonylation. The extent of observed oxidative damage was correlated with accumulation of Th in examined organs and further associated with histological alterations. Furthermore it was found that these effects were significantly lower when the chelating agent, Ca-DTPA, was given 1 h after Th injection. CONCLUSION: Administration of subtoxic concentration of Th to mice markedly altered the liver functions and induced oxidative stress in the liver, femur and spleen of mice. The results further demonstrated that Ca-DTPA significantly protected mice against the toxic effects of Th. PMID- 18386200 TI - Introduction: issues and viewpoints in pediatric psychopharmacology. PMID- 18386201 TI - An international perspective on pediatric psychopharmacology. AB - The use of pharmacotherapy for children and adolescents with mental disorders varies widely across countries. More than 80% of the world use of stimulant medications occurs in the USA. The use of antidepressants and antipsychotics is many times greater in the USA than in other countries. Factors likely to influence the pediatric use of psychotropic medications are here examined and discussed. Variability in use reflects differences in diagnostic systems, clinical practice guidelines, drug regulation, health services organization, availability and allocation of financial resources, and cultural attitudes towards childhood behavioral and emotional disturbances. Cultural context seems to exert a greater influence on the identification and management of psychiatric disorders than on other areas of medicine. It is currently unknown if the heterogeneity in treatment approaches results in differential clinical outcomes and prognosis. A better understanding of the factors underlying international variability may help clarify the process of diagnosis and treatment selection in child and adolescent psychiatry. PMID- 18386202 TI - How to manage conflicts of interest with industry? AB - The use of medications has risen steadily in psychiatry. Perhaps in response, during the past few years there has been increasing scrutiny of alleged unethical behaviours by medical researchers, educators, and practitioners secondary to influence by the pharmaceutical industry. Research is quite consistent that gifts and generous financial arrangements can dampen skepticism, sometimes unconsciously, and thereby persuade recipients to advocate for or prescribe medications that are more expensive, but no more effective, than alternatives. Interestingly, this research-backed premise that physicians can be lured by gifts remains often disbelieved by recipients. Adding to such inducements to prescribe new, expensive medications are pressures from patients due to the increasing ubiquity of direct-to-consumer advertising. Criticism from patient advocate groups, government agencies, and the press has sparked debate within the profession. Many medical journals, academic medical centre research and educational endeavours, and medical organizations are reviewing policies to eliminate, or better manage, their conflicts of interest with industry. The basic ethical standard is that although pharmaceutical companies' primary concern is for its shareholders, physicians' primary concern must be for their patients. PMID- 18386203 TI - Parental attitudes and involvement in psychopharmacological treatment for ADHD: a conceptual model. AB - Current evidence strongly points to significant parental concerns about exposing their children to psychopharmacological medications. This is true even with stimulant medications for attention deficit/hyperactivity disorder (ADHD), an evidence-based treatment for children. Theory and prior evidence would predict that parents' hesitancy to give their children psychopharmacological medication is likely due to what they hear from family and friends about medication's untoward effects. That social networks impact problem definition, treatment evaluation, and service use is well established. However, it is not known how parents make treatment decisions as they cope with the reality that their child might benefit from psychopharmacological medication. The qualitative research described in this paper addresses this gap. A conceptual model is presented that describes parents' treatment decisions and participation in their children's care as they shift from parenting a child with behavioural problems to parenting a child who eventually requires psychopharmacological medication. Further research is needed to evaluate how this model can offer a more comprehensive understanding of how parents respond to a diagnostic label and various treatment options. It is hoped that this research will stimulate innovative approaches for tailoring treatments that are respectful of the individual needs of children and their families. PMID- 18386204 TI - Beyond the 'typical' patient: treating attention-deficit/hyperactivity disorder in preschoolers and adults. AB - Attention-deficit/hyperactivity disorder (ADHD), typically affects school-aged children, but can present during the preschool years and persist into adulthood. Accurate diagnosis for preschoolers and adults requires adaptation of the current diagnostic criteria to account for differences in symptomatology across the age span. The differential diagnosis of ADHD and the pattern of psychiatric comorbidity vary with each age group and complicate diagnosis and management. To maximize outcomes clinicians must be able to accurately identify ADHD across the lifespan, and develop comprehensive, collaborative treatment plans. The Preschool ADHD Treatment Study (PATS) demonstrated the potential utility of methylphenidate for treating ADHD in preschoolers, and trials of psychostimulants and atomoxetine have shown some benefits for adults. Behavioural interventions likely have an adjunctive role in ADHD treatment for both groups. More research, however, is needed to determine the safest and most effective pharmacotherapies and psychosocial interventions for these non-typical patients. PMID- 18386205 TI - Pharmacological treatment of pathologic aggression in children. AB - Children with psychiatric illness may display pathologic aggression (PA) that is destructive, severe, chronic, and unresponsive to psychosocial and psychopharmacological treatment of their underlying condition(s) and psychosocial interventions specifically targeting PA. For this subset of children with PA, pharmacotherapy may be an appropriate treatment option to optimize their functioning. This article reviews pharmacological treatment studies for PA in children and the safety and efficacy of risperidone, olanzapine, lithium, divalproex sodium, methylphenidate, and typical antipsychotics in this patient population. While safety needs to be emphasized when prescribing medication for these patients, serious health and safety risks are also raised when PA is not effectively treated. Future research is needed to evaluate whether the long-term risks associated with the pharmacological treatment of PA outweigh the potential benefits to the child. PMID- 18386206 TI - Combining and sequencing medication and cognitive-behaviour therapy for childhood anxiety disorders. AB - Despite the absence of data on the efficacy of combination therapy (i.e., psychosocial and medication) for the treatment of anxiety disorders in youths, clinicians in clinical practice often utilize this treatment approach. This paper discusses issues related to sequencing, combining, and integrating cognitive behavioural and pharmacological interventions for anxiety disorders in children and adolescents. We briefly summarize the empirical evidence for mono and combination therapy and raise a variety of issues that should be considered when making treatment decisions. Finally, we present an integrated treatment model to facilitate the delivery of a comprehensive treatment approach across care providers. These suggestions are geared toward optimizing clinical outcomes for anxious youths. PMID- 18386207 TI - Autism: the role of cholesterol in treatment. AB - Cholesterol is essential for neuroactive steroid production, growth of myelin membranes, and normal embryonic and fetal development. It also modulates the oxytocin receptor, ligand activity and G-protein coupling of the serotonin-1A receptor. A deficit of cholesterol may perturb these biological mechanisms and thereby contribute to autism spectrum disorders (ASDs), as observed in Smith Lemli-Opitz syndrome (SLOS) and some subjects with ASDs in the Autism Genetic Resource Exchange (AGRE). A clinical diagnosis of SLOS can be confirmed by laboratory testing with an elevated plasma 7DHC level relative to the cholesterol level and is treatable by dietary cholesterol supplementation. Individuals with SLOS who have such cholesterol treatment display fewer autistic behaviours, infections, and symptoms of irritability and hyperactivity, with improvements in physical growth, sleep and social interactions. Other behaviours shown to improve with cholesterol supplementation include aggressive behaviours, self-injury, temper outbursts and trichotillomania. Cholesterol ought to be considered as a helpful treatment approach while awaiting an improved understanding of cholesterol metabolism and ASD. There is an increasing recognition that this single-gene disorder of abnormal cholesterol synthesis may be a model for understanding genetic causes of autism and the role of cholesterol in ASD. PMID- 18386208 TI - Mood lability and bipolar disorder in children and adolescents. AB - Pediatric bipolar disorder, once considered rare, has reportedly increased in incidence over the past 10 years. There is significant debate about the phenomenology, diagnosis and treatment of this illness. The diagnostic assessment of children with severe mood and behavioural disturbance is of considerable public health importance as the ultimate diagnosis can have significant treatment implications and can impact the level of stigma experienced by children and their families. The purposes of this paper are to: 1) review current issues in the phenomenology and diagnostic assessment of bipolar disorder in children and adolescents; 2) review recent research suggesting that youths with a chronic course of illness should be considered a separate group from those with an episodic course; and 3) offer suggestions for future studies to address the various phenomenological controversies. PMID- 18386209 TI - Can randomized controlled trials be done with suicidal youths? AB - Designing a randomized controlled trial (RCT) to identify the efficacy of treatments for suicidal youths is challenging and requires great care. The lack of a comprehensive treatment evidence base for suicidal youths is largely the result of the complexity of these youths and the many impediments to mounting a RCT for the commonly used treatments-medication and psychotherapy. Correctly choosing the hypothesis to be tested, the appropriate sampling frame, the intervention and control groups, and the outcome variables increases the likelihood that the study will be feasible, ethical and clinically meaningful. To meet these goals, the sampling frame needs to be broad, but not require overly complex assessment, treatment and rescue protocols; intervention and control groups must meet or beat the 'standard of care', and be acceptable to participants and clinicians. The outcome variables need to be measurable, sensitive to change and common enough to identify between group differences without large samples. This manuscript: (1) reviews existing studies of interventions for suicidal youths, (2) describes challenges in selecting the population, treatment and control conditions, and appropriate and meaningful outcome measures, and (3) discusses alternative methodological approaches that may be necessary to address the complex treatment needs of suicidal youth. PMID- 18386210 TI - Medication management of pediatric eating disorders. AB - This article provides an overview of psychopharmacological treatments for pediatric eating disorders (EDs). Although EDs usually begin in adolescence, there are few pharmacological treatment trials specific to this age group and a paucity of controlled data. Empirical evidence suggests that psychological, behavioural and family interventions should be the primary modalities of treatment for these conditions. In severely underweight patients behavioural weight restoration should be attempted before pharmacological intervention, especially since starvation is known to aggravate obsessional and depressive symptomatology. Evidence from controlled trials supports the use of antidepressants for the treatment of bulimia nervosa (BN) in adults; however, similar studies have not yet been performed in youths. For anorexia nervosa (AN), there are no pharmacotherapies of proven efficacy in either adults or youths. Nonetheless, clinical experience and uncontrolled evidence suggests that some children and adolescents may benefit from thoughtful use of psychotropic medications on an individual basis in the context of a multimodal treatment plan. Regarding binge eating disorder (BED), adult literature shows positive short-term effects on binge eating for both pharmacological (especially selective serotonin reuptake inhibitors) and behavioural interventions, but unclear effects on weight. Clearly, psychopharmacological interventions for pediatric EDs would benefit from more research. PMID- 18386211 TI - Do all obsessive-compulsive disorder subtypes respond to medication? AB - Obsessive-compulsive disorder (OCD) in children is strikingly similar in clinical presentation and treatment responsiveness to OCD in adults. While treatment is commonly effective for OCD not all subtypes of OCD are similarly responsive to treatment. Numerous studies describe the differential responsiveness of OCD subtypes to pharmacological treatment in adults, yet few such studies exist in pediatric OCD. This manuscript reviews the extant literature that addresses differential response of OCD subtypes to medication intervention. Specific OCD subtypes can be derived utilizing the following strategies: symptom factor analysis, comorbidity latent class analysis, concurrent internalizing disorders, concurrent externalizing disorders, tic-related subtype and early-onset subtype are reviewed in relation to data on differential pharmacotherapy response. Only externalizing disorders moderate treatment response in pediatric OCD. Specifically, attention-deficit hyperactivity disorder, oppositional defiant disorder and conduct disorder are associated with poorer response to medication treatment. Hoarding appears to be associated with a poor response to medication in adults, but data are sparse in children. The presence of tic disorders may also predict poorer response to pharmacotherapy in pediatric OCD. Strategies for treatment of the tic-related subtype of OCD, while strongly supporting the use of antipsychotic-augmentation for enhanced response in adults, are lacking controlled data in pediatric OCD. PMID- 18386212 TI - Monitoring and management of antipsychotic-related metabolic and endocrine adverse events in pediatric patients. AB - Efficacy differences among antipsychotics are relatively small and difficult to predict, however, differences are generally larger and more predictable regarding adverse effects. Antipsychotic related metabolic and endocrine abnormalities are most worrisome, as they are risk factors for cardiovascular morbidity and mortality. Importantly, children and adolescents may be more sensitive to antipsychotic related adverse events than adults, including developmentally inappropriate weight gain and glucose, lipid and prolactin abnormalities, which can be associated with adverse physical outcomes during childhood. Careful selection of an antipsychotic, as well as proactive monitoring and management of side effects should be part of routine clinical practice. Because children and adolescents undergo expected physical changes, it is important to use developmentally appropriate thresholds for weight and metabolic measures. Research is required to elucidate risk factors, mechanisms and best interventions for antipsychotic related adverse endocrine and metabolic effects in pediatric patients. PMID- 18386213 TI - SSRI adverse events: how to monitor and manage. AB - Antidepressants are efficacious for pediatric major depressive disorder, obsessive compulsive disorder (OCD), and non-OCD anxiety disorders. Antidepressants should be used in an evidence-based fashion, with frequent monitoring for side effects, especially when initiating treatment and adjusting dosage. With diligence to appropriate prescribing and monitoring, the benefits of therapy outweigh the potential of treatment related risk. PMID- 18386214 TI - The contribution of pharmacoepidemiology to the antidepressant-suicidality debate in children and adolescents. AB - A number of concerns have recently been raised about whether or not antidepressant medications are associated with suicidal thoughts and behaviour in children and adolescents. These concerns are based largely on results of meta analyses of randomized, controlled trials (RCTs). Controversy exists about generalizing evidence from short-term RCTs, designed primarily to test efficacy outcomes, to routine practice settings. Pharmacoepidemiological studies complement RCTs by using observational methods to examine safety and effectiveness of medications in the general population. This article reviews the contribution of pharmacoepidemiology to the controversy surrounding suicide risk in children and adolescents taking antidepressants, noting how variations in study design and adjustment for potential confounding factors influence outcome. PMID- 18386216 TI - Real-time contrast imaging: a new method to monitor capillary recruitment in human forearm skeletal muscle. AB - OBJECTIVE: Muscle capillary perfusion can be measured by contrast-enhanced ultrasound. We examined whether a less time-consuming ultrasound technique, called "real-time imaging," could be used to measure capillary recruitment in human forearm skeletal muscle. METHODS: We measured microvascular blood volume and microvascular flow velocity using bolus injections of contrast microbubbles after forearm muscle exercise and a two-hour infusion of insulin into the brachial artery (both associated with capillary recruitment) and after sodium nitroprusside infusion (no changes in flow distribution). RESULTS: After an intravenous bolus injection of the contrast agent, the steady-state concentration of contrast agent in forearm muscle lasted long enough (approximately 190 seconds) for the duration of the measurements (which take 70-80 seconds), rendering the continuous infusion of microbubbles unnecessary. Microvascular blood-volume measurements showed a good short-time reproducibility and a good reproducibility after repositioning of the forearm. Reproducibility of microvascular flow velocity was too low. Exercise and insulin infusion both increased microvascular blood volume, consistent with capillary recruitment. Sodium nitroprusside had no effect. CONCLUSION: Real-time contrast imaging, after bolus injections of an ultrasound contrast agent, provides reliable information about capillary recruitment in human forearm skeletal muscle, and may offer a valuable tool in studying human (patho)physiology. PMID- 18386215 TI - Effects of old age on vascular complexity and dispersion of the hepatic sinusoidal network. AB - OBJECTIVES: In old age, there are marked changes in both the structure of the liver sinusoidal endothelial cell and liver perfusion. The objective of this study was to determine whether there are also aging changes in the microvascular architecture and vascular dispersion of the liver that might influence liver function. METHODS: Vascular corrosion casts and light micrographs of young (4 months) and old (24 months) rat livers were compared. Fractal and Fourier analyses and micro-computed tomography were used. Vascular dispersion was determined from the dispersion number for sucrose and 100-nm microspheres in impulse response experiments. RESULTS: Age did not affect sinusoidal dimensions, sinusoidal density, or dispersion number. There were changes in the geometry and complexity of the sinusoidal network as determined by fractal dimension and degree of anisotropy. CONCLUSIONS: There are small, age-related changes in the architecture of the liver sinusoidal network, which may influence hepatic function and reflect broader aging changes in the microcirculation. However, sinusoidal dimensions and hepatic vascular dispersion are not markedly influenced by old age. PMID- 18386217 TI - Longitudinal and radial gradients of PO(2) in the hamster cheek pouch microcirculation. AB - OBJECTIVES: The aim of this study was to determine longitudinal and radial gradients in oxygen tension (PO(2)) in microvessels of the hamster cheek pouch. METHODS: We measured PO(2) using the phosphorescence-quenching method in two orders of arterioles (45.8 +/- 5.5 and 19.9 +/- 1.8 micro m diameter), capillaries, and two orders of venules (50.5 +/- 3.4 and 21.4 +/- 2.0 micro m diameter) in order to determine the longitudinal PO(2) gradient. At the arteriolar and venular sites, we also measured PO(2) at four different sites for an analysis of radial PO(2) gradients: centerline, inside wall (larger arteriole and venule only), outside wall, and interstitium. We used 10 hamsters weighing 115 +/- 27 g anesthetized with pentobarbital intraperitoneally and maintained with alpha-chloralose intravenously. The cheek pouch was everted and a single layered preparation was studied by intravital microscopy. Albumin-bound Pd porphyrin was infused into the circulation and excited by flash illumination at 10 Hz, with a rectangular diaphragm limiting the excitation field to 5 x 25 micro m. RESULTS: In the longitudinal direction, intravascular PO(2) decreased significantly (P < 0.01) from large arterioles (39.5 +/- 2.3 mmHg) to small arterioles (32.2 +/- 0.3 mmHg), then to capillaries (30.2 +/- 1.8 mmHg), and on to small venules (27.3 +/- 2.1 mmHg) and large venules (25.5 +/- 2.2 mmHg). In the radial direction, PO(2) decreased significantly (P < 0.01) in and around larger arterioles, and to a lesser extent, around the smaller ones (P < 0.05). There was no significant PO(2) gradient, longitudinal or radial, associated with venules. The PO(2) difference from the centerline to the outside wall in large arterioles was 8.3 +/- 1.4 mmHg, and most of the decline in PO(2) in the radial direction was contributed by the intravascular difference (4.7 +/- 2.1 mmHg) and only about 1.0 +/- 2.7 mmHg by the transmural difference. CONCLUSIONS: Our data show that there are large intra-arteriolar radial PO(2) gradients, but no large transmural PO(2) differences, suggesting that the oxygen consumption of the microvessel wall is not exceptionally high. PMID- 18386218 TI - Age-related alterations in reactivity of cerebral arterioles: role of oxidative stress. AB - OBJECTIVE: Our goal was to identify the role of oxidative stress via activation of NAD(P)H oxidase in cerebrovascular dysfunction in aged rats. METHODS: We examined the reactivity of cerebral arterioles in adult and aged Fisher-344 rats to endothelial nitric oxide synthase (eNOS)-dependent (acetylcholine and adenosine diphosphate [ADP]) and-independent (nitroglycerin) agonists before and during application of tempol, apocynin, and diphenyleneiodonium chloride (DPI). We used Western blot to examine subunits of NAD(P)H oxidase, eNOS, and superoxide dismutase (SOD-1) in cerebral microvessels and parietal cortex. Finally, we measured superoxide production by cortex tissue in adult and aged rats. RESULTS: Acetylcholine-and ADP-induced, but not nitroglycerin-induced, dilatation of cerebral arterioles was impaired in aged compared to adult rats. While tempol, apocynin, and DPI did not alter responses in adults, they alleviated impaired eNOS-dependent vasodilatation in aged rats, without influencing responses to nitroglycerin. eNOS and p67phox proteins were increased in cerebral microvessels from aged compared to adult rats. Further, p67phox and gp91phox proteins were increased, but SOD-1 protein was decreased, in cortex tissue of aged rats. Basal and agonist-induced production of superoxide was elevated in aged rats. CONCLUSIONS: Aging impairs eNOS-dependent reactivity of cerebral arterioles via an increase in superoxide produced by activation of NAD(P)H oxidase. PMID- 18386219 TI - Monitoring the capillary-alveolar leakage in an A.R.D.S. model using broncho alveolar lavage. AB - OBJECTIVES: We developed a modified broncho-alveolar lavage (BAL) technique in order to perform repeated measurements of capillary-alveolar leakage of a macromolecule in oleic acid (OA)-induced lung injury. METHODS: BAL was performed in anesthetized dogs in a closed lung sampling site, using a bronchoscope fitted with an inflatable cuff. Fluorescein-labeled Dextran (FITC-D70) was continuously infused and its concentration measured in plasma and BAL fluid. A two-compartment model (blood and alveoli) was used to calculate K(AB), the transport-rate coefficient of FITC-D70 from blood to alveoli. K(AB) was estimated every 15 minutes over three hours. RESULTS: K(AB), close to zero at base-line both in control (n = 3) and in OA-injured lungs (n = 7), reached a peak in permeability 30 minutes after the induction of OA injury (K(AB) = 1.43 +/- 0.31 . 10(- 3) . minutes(- 1)), followed by a slow recovery. CONCLUSIONS: We conclude that this technique allows the monitoring of capillary-alveolar transport of macromolecules in in vivo experimental models. This monitoring may prove useful to study the mechanisms of the exudative stage of acute lung injury and to test therapies aimed at slowing the alveolar accumulation of plasma proteins and procoagulant factors that contribute to alveolar fibrosis. PMID- 18386220 TI - Xanthine oxidase interaction with vascular endothelial growth factor in human endothelial cell angiogenesis. AB - OBJECTIVES: Reduced capillary density occurs early in cardiovascular diseases. Oxidant stress is implicated in endothelial apoptosis. We investigated the effects of xanthine oxidase (XO) on endothelial survival signaling: protein kinase B/Akt, its cross-talk with p38 MAPK and apoptosis pathways, and its effect on vascular tube formation in vascular endothelial growth factor (VEGF)-simulated human umbilical vein cells. METHODS: We studied primary cultured human endothelial cells from the umbilical cord. Reactive oxygen species (ROS) production was detected by dihydroethidium staining, cell-signaling pathways by western blots, cell survival by western blots, and nuclear chromatin and angiogenesis response by MTT proliferation assay and three-dimensional Matrigel cultures. RESULTS: Exogenous XO increased cellular ROS production and caused superoxide-dependent inhibition of Akt phosphorylation and enhancement of p38 MAPK phosphorylation in a time-and dose-dependent manner. In contrast, application of the XO inhibitor oxypurinol or allopurinol inhibited VEGF stimulated Akt phosphorylation, indicating that endogenous XO promotes VEGF induced endothelial cell (EC) survival signaling. Exogenous XO induced activation of caspase-3 and reduced expression of the anti-apoptosis protein Bcl-2. Exogenous XO also reduced EC viability, proliferation, and vascular tube formation by p38 MAPK-dependent, phosphoinositide 3-kinase (PI3-K) reversible mechanisms; whereas VEGF promoted EC survival by PI3-K-dependent, p38 MAPK independent effects. CONCLUSIONS: Exogenous XO activity is an important contributor to endothelial mechanisms for microvascular rarefaction, by modulation of cell survival signaling pathways; however, endogenous XO is necessary for maintaining EC survival. PMID- 18386221 TI - A fight at the opera. PMID- 18386222 TI - Identifying autoantigens as theranostic targets: antigen arrays and immunoproteomics approaches. AB - The development of theranostics holds great potential for the advancement of personalized medical treatments for human diseases. The theranostic approach combines diagnostic testing with predicted treatment outcomes in order to specifically tailor medical treatments for individual patients. As such, theranostic tests have the potential to revolutionize the diagnosis, treatment and monitoring of human diseases. Currently, the search is underway to identify novel targets for use in the development of theranostic approaches. The aims of this review are: to briefly describe the current capabilities of antigen autoantibody microarrays, including our native antigen 'reverse-capture' autoantibody microarray platform to identify theranostic targets; to detail other immunoproteomics approaches for the identification of theranostic targets, including serological analysis of antigens by recombinant complementary (c)DNA expression cloning, serological proteome analysis, and autoantibody-mediated identification of antigens; and to provide examples of their use in theranostic discoveries. PMID- 18386223 TI - Membrane proteins as diagnostic biomarkers and targets for new therapies. AB - Membrane proteins, especially plasma membrane proteins, form one of the most interesting classes of proteins among disease biomarker candidates. Because of their localization on the surface of cells and organelles, membrane proteins also represent potential drug targets. In this review, developments in the characterization of membrane proteins and their role in the treatment of disease, in particular cancer treatment, are presented. PMID- 18386224 TI - EGFR-targeted therapy in colorectal cancer: does immunohistochemistry deserve a role in predicting the response to cetuximab? AB - The EGF receptor (EGFR) has emerged as a rational target for anticancer therapy for the treatment of colorectal cancer (CRC). Positive immunohistochemistry (IHC) staining for EGFR is used as a criterion for patient selection; however, doubt has been cast on the utility of this method. Not only is the response to cetuximab, an anti-EGFR mAb, low in patients expressing EGFRs, but a similar response to cetuximab has also been described in patients who do not express EGFRs. This review aims to evaluate the possible cause of the lack of correlation between the efficacy of cetuximab and EGFR IHC staining in CRC, as well as any modifications in the IHC method necessary to optimize patient selection for cetuximab therapy. In our opinion, the heterogeneous expression of the receptor in the neoplastic population and the inability of the mAbs used to predict the response to cetuximab could be the major cause of the failure of IHC staining as a reliable tool for patient selection. The use of specific mAbs directed against the phosphorylated and mutant form (EGFRvIII) of the EGFR could reinstate IHC as a valid predictor of response to therapy. PMID- 18386225 TI - Multifunctional peptide-based nanosystems for improving delivery and molecular imaging. AB - Nanotechnology is a multidisciplinary scientific field impacting on various facets of preclinical and clinical research. One of the areas most affected is the development of medical treatments, that is, nanomedicine. In particular, drug delivery is expected to benefit from the unique ability of nanoparticles to selectively target different tissues and cellular compartments. In this review, advances using nanoparticles to improve the delivery of peptides by rendering them more stable, less susceptible to degradation, and able to cross physical barriers that typically restrict circulatory system access are discussed. In addition, the use of peptide-nanoparticle conjugates to target normal and diseased tissues to deliver therapeutic agents or to image them for diagnostic and screening purposes, or both, are highlighted. In the next few years these technologies are expected to allow control of drug delivery, targeting, and imaging that is currently unattainable. PMID- 18386226 TI - The therapeutic potential of antisense-mediated exon skipping. AB - Antisense-mediated exon skipping is an approach that uses antisense oligonucleotides (AONs) to modulate splicing by hiding specific sites essential for exon inclusion from the splicing machinery. AONs that block aberrant splice sites can restore normal splicing, whereas AONs targeting alternative splice sites can switch splicing patterns from detrimental to beneficial isoforms or produce non-functional mRNAs that lead to gene knockdown. Furthermore, AONs have also been used to restore a disrupted reading frame, thereby generating semi functional proteins instead of non-functional proteins. Proof-of-concept has been obtained for each of the above AON applications in vitro and for some applications in vivo. Antisense-mediated reading frame restoration is the most promising therapy for Duchenne muscular dystrophy. Data from a first clinical trial are encouraging and additional trials are ongoing or are expected to be initiated soon. PMID- 18386227 TI - Emerging role of microRNAs in disease pathogenesis and strategies for therapeutic modulation. AB - Micro (mi)RNAs are small, regulatory RNA molecules that are integral components of the genetic program in the majority of cells. They are thought to regulate up to one third of all human genes and have been linked to critical processes in disease pathogenesis. The diverse role of miRNAs in disease pathogenesis suggests that the modulation of miRNA function by utilizing techniques such as the use of antagomirs, locked nucleic acids, or miRNA sponges may produce novel therapeutic approaches. In this review, the current understanding of post-transcriptional gene regulation by miRNAs is discussed and insights into the function of miRNAs in tumorigenesis, immune responses, muscle function, organogenesis, and cell lineage decisions are reviewed. PMID- 18386228 TI - RNAi therapeutics: an update on delivery. AB - RNA interference (RNAi) has rapidly advanced from a laboratory observation into a major area of research within biology and medicine. RNAi is triggered by short interfering RNAs (siRNAs) of between 19 and 21 nucleotides in length, which induces the targeted cleavage of mRNA with sequences of homology to the siRNA. Because of its high degree of specificity and efficacy, the potential for RNAi based therapeutics was recognized at an early stage. However, development of RNAi based agents has been hindered because siRNAs are unstable in serum and delivery across the cell membrane is highly inefficient. Numerous methods have been developed to facilitate delivery of RNAi in animals and patients, each with their own set of advantages and disadvantages. This review discusses publications between 2005 to 2007 in the area of RNAi delivery, with a particular focus on in vivo application and clinical trials. PMID- 18386229 TI - Sugar boost: when ribose modifications improve oligonucleotide performance. AB - Since the first antisense oligonucleotides were described, synthetic oligonucleotides have been used to modulate gene expression for numerous research and therapeutic purposes over the last 30 years. In order to improve the performance of oligonucleotides in therapeutic applications, various chemical modifications of nucleotides have been developed and applied to antisense, antigen, aptamer and short interfering RNA (siRNA) therapeutics. For example, backbone substitution of phosphodiester bonds to create DNA phosphorothioates and 4'-thio RNA analogs, increases resistance to nuclease degradation. Modifications to improve the activity of oligonucleotides by 2'-O-modification of the ribose sugar, and the systematic approaches used to evaluate the potential of mixed oligonucleotides, are discussed in this review. Although structural data from crystallization studies appear to confirm observed gains in thermodynamic stability with 2'-O-modifications, no simple rules can be drawn to predict the overall effect of individual modifications for the rational design of mixed oligonucleotides. PMID- 18386230 TI - VB4-845, a conjugated recombinant antibody and immunotoxin for head and neck cancer and bladder cancer. AB - Viventia Biotech Inc, under license from the University of Zurich, is developing VB4-845, comprising a Pseudomonas exotoxin fused to an anti-epithelial cell adhesion molecule single-chain antibody fragment, for the potential treatment of head and neck cancer (intratumoral) and bladder cancer (intravesical). VB4-845 is currently undergoing phase II and III clinical trials in patients with head and neck squamous cell carcinomas, as well as phase II clinical trials for the treatment of superficial transitional cell carcinoma of the bladder. PMID- 18386231 TI - Sitimagene ceradenovec, a gene therapeutic for the treatment of glioma. AB - Ark Therapeutics Group plc is developing sitimagene ceradenovec, a herpes simplex virus thymidine kinase/ganciclovir gene therapy utilizing an adenoviral vector, for the potential treatment of glioma. Sitimagene ceradenovec is currently undergoing a phase III clinical trial in malignant glioma patients. PMID- 18386232 TI - Starting a liver transplant program in Turkey; Akdeniz University model. PMID- 18386233 TI - Metastatic/resected lymph nodes ratio-based classification in gastric cancer. AB - BACKGROUND/AIMS: Many studies have shown that the metastatic lymph node ratio, calculated by dividing the number of metastatic lymph nodes by the total number of lymph nodes, is an important prognostic factor in gastric cancer. In the present study, the applicability of the metastatic in the 1997 Tumor Node Metastasis system was investigated using our clinical data and discussed in light of the literature. METHODS: The study was performed on the 166 patients with gastric cancer in whom R0 resection could be performed and more than 15 nodes were resected. The possible effects of age, gender, location, type of resection, number of resected lymph nodes, depth of invasion, number of involved lymph nodes, tumor grade and metastatic on survival were analyzed. RESULTS: There was a direct correlation between the total number of nodes and the number of metastatic nodes (r: 0.319, p<0.0001). However, there was no correlation between metastatic and the total number of nodes (r: 0.0072, p: 0.354). Tumor location, size, depth of invasion, number of involved nodes and metastatic were found to be determinants of survival in univariate analysis. Cox regression analysis identified metastatic as the only independent prognostic factor. CONCLUSIONS: A new staging system based on metastatic will be resistant to stage migration and will include the surgical approach in staging. However, further studies are required to determine appropriate cutoff values and the best approach to patients with less than 15 resected nodes. PMID- 18386234 TI - Comparison of the effects of intravenous midazolam alone and in combination with meperidine on hemodynamic and respiratory responses and on patient compliance during upper gastrointestinal endoscopy: a randomized, double-blind trial. AB - BACKGROUND/AIMS: We examined hemodynamic responses during gastroscopy in healthy subjects and compared the changes with midazolam alone versus in combination with meperidine. The aim of this study was to evaluate if either method had any advantages or disadvantages with respect to patient compliance and the commonly seen side effects. METHODS: Thirty patients who were otherwise healthy were included in each group. Either midazolam 0.05 mg/kg IV (Group I) or meperidine 0.3 mg/kg IV followed by midazolam 0.05 mg/kg (Group II) IV were used for sedation. Data of noninvasive hemodynamic and cardiac parameters were recorded before and at the 1st minute after medication, and at the 1st minute and 2-min intervals during the procedure. Endoscopists assessed the comfort of patients according to pre-determined criteria. Statistical analysis was performed for both inter-group and in-group comparisons of parameters. RESULTS: Heart rate increased significantly in Group I (p<0.05). Blood pressures and oxygen saturation decreased significantly with sedation in both groups during endoscopy (p<0.05), without significant difference between the groups for the changes in these parameters (p>0.05). Patient compliance was significantly better in Group II than in Group I, for all measured criteria. CONCLUSIONS: We observed that heart rate increases significantly whereas SAP, DAP and SpO2 decrease significantly with both sedation methods. Groups did not differ except for the significantly higher increase in heart rate in Group I. Patient compliance was significantly better with combined sedation. We believe that combined sedation in selected patients provides a safe sedation with a mild to moderate increase in heart rate and a better patient compliance during gastroscopy. PMID- 18386235 TI - Human tissue transglutaminase antibody screening by immunochromatographic line immunoassay for early diagnosis of celiac disease in Turkish children. AB - BACKGROUND/AIMS: Celiac disease has a large prevalence worldwide. There are a limited number of comparable epidemiological data for celiac disease in Turkey. The aim of this preliminary study was to determine the prevalence of celiac disease in a sample of 1000 Turkish children by a novel, simple, and visual one step immunoassay screening test. METHODS: This prospective study consisted of 1000 serum samples from apparently healthy children and children with disorders other than celiac disease aged between 2-18 years who presented as outpatients at Ankara University, Faculty of Medicine, Department of Pediatrics. Sera were tested for IgA-class antibodies against human tissue transglutaminase and gliadin by rapid immunochromatographic line immunoassay. Endomysial antibody IgA against human tissue transglutaminase and AGA IgA/IgG were also tested by ELISA as a second step when the result of the screening test was positive. Small bowel biopsy was recommended to all the children with positive anti-tissue transglutaminase and/or endomysial antibody results. RESULTS: Ten of the 1000 individuals (1%) had positive antibody screening test to human tissue transglutaminase. All tissue transglutaminase-positive samples revealed good correlation with endomysial antibody by ELISA method. Subsequently small bowel biopsy was performed in all serology-positive cases. Biopsy results confirmed a diagnosis of celiac disease in nine cases. The prevalence of biopsy- proven celiac disease was 1:111 (0.9%). CONCLUSIONS: Determination of anti-tissue transglutaminase antibodies by simple visual system for celiac disease appeared to be as reliable as the ELISA system. It is easy to perform and interpret, cost effective, and rapid, as pointed out in other previous studies, as a screening test in large population-based studies. The prevalence of celiac disease in the overall sample of Turkish children (1:111 or 0.9%) in this preliminary study is similar to that reported in European and Middle Eastern countries and the United States. PMID- 18386236 TI - Relationship between overexpression of ras p21 oncoprotein and K-ras codon 12 and 13 mutations in Turkish colorectal cancer patients. AB - BACKGROUND/AIMS: The activation of ras family genes plays an important role in colorectal tumorigenesis. We investigated the clinicopathological characteristics and point mutations of K-ras oncogene codons 12/13 and ras p21 expression using paraffinembedded materials from cancerous and the surrounding normal tissues of 53 colorectal cancer cases. METHODS: K-ras codons 12 and 13 point mutations were analyzed by PCR-Single- Strand Conformational Polymorphism (SSCP) and followed by DNA sequencing, while ras p21 expression was evaluated using immunohistochemistry. RESULTS: Mutations of K-ras and overexpression of the ras p21 were detected in 11% and 76% of the tumors, respectively. Ras protein level in tumor was increased an average of 4.6-fold over that of normal mucosa. Ras p21 overexpression did not correlate with any of the clinicopathological parameters examined. K-ras gene mutations were found mostly in the presence of a mucinous component within the tumor (p=0.06). Follow-up data were available for 43 patients. There was no statistically significant correlation between these alterations and patient outcomes. CONCLUSIONS: Our data suggest that, apart from K-ras codons 12/13 point mutations, overexpression of the ras family genes is important in the development of the disease but it appears not to be predictive of survival. Furthermore, mucinous secretion in the colorectum may represent a distinct genetic pattern. PMID- 18386237 TI - Evaluation of the first 100 liver transplantations. AB - BACKGROUND/AIMS: We aimed to present the experience of the first 100 liver transplantations carried out at Akdeniz University. METHODS: The data of 100 patients in pediatric and adult age groups who underwent liver transplantation at Akdeniz University Organ Transplantation Center between January 2000 and January 2007 were examined retrospectively. The cases between January 2000 and December 2003 were evaluated as the first term and those between January 2004 and January 2007 as the second term. RESULTS: The mean age of the 100 patients (52M, 48F) was 38.6+/-17.3 (1-68) years. One-year and three-year survival rates of the patients were determined as 67.3% and 54.3% in the first term and 88.7% and 79.3% in the second term, respectively. CONCLUSIONS: With better comprehension of recipient and donor surgery technique, in addition to accumulation of knowledge and experience, the results in liver transplantation might be improved. PMID- 18386238 TI - The optimal treatment of hydatid cyst of the liver: radical surgery with a significant reduced risk of recurrence. AB - BACKGROUND/AIMS: The management of hydatid cyst of the liver, which is still endemic in Turkey, varies from medical treatment or percutaneous drainage to different surgical procedures. In this study, we aimed to compare the efficacy of radical surgical procedures and conservative interventions with respect to recurrence rates. METHODS: Patients who underwent any type of surgical treatment between March 1994 and March 2007 due to liver cyst hydatid were retrospectively evaluated. Data collection included demographic variables, diagnostic methods, surgical procedures, and morbidity and mortality rates. RESULTS: 242/258 (93.8%) patients with liver hydatid cyst underwent surgery, and the characteristics of 221 (91.3%) (123 female, 98 male) of these patients matched the criteria of the study. The mean age of the patients was 51 years (18-82 years). The diagnostic methods primarily included abdominal ultrasonography and computed tomography with a rate of 61.8% and magnetic resonance imaging in 12% of the patients. The patients were divided into two groups with respect to the treatment modality: Group A (n=92) - radical surgical treatment and Group B (n=129) - conservative surgery. The overall rate of recurrence was 15.3%. In Group B, this rate was 24% (n=31), whereas only 3.2% of the patients (n=3) in Group A had recurrence in the follow-up (p<0.05). The morbidity rate of the patients who underwent radical surgical modalities was also significantly lower. CONCLUSIONS: Although conservative surgical procedures are considered simpler and safer to perform, the rate of postoperative complications such as biliary fistula, residual cavity and recurrence, and cavity suppuration has been reported to be about 35%. On the other hand, radical surgery can be performed with low risk of recurrence (3.2%). We believe radical surgical procedures present a lower rate of recurrence and less morbidity, and thus should be the surgical treatment of choice for hepatic hydatid disease. PMID- 18386239 TI - Retrorectal tumors: a case series. AB - BACKGROUND/AIMS: Retrorectal masses are rarely encountered in surgical practice, and they arise from congenital remnants, and from osteogenic, neurogenic, inflammatory, or metastatic processes. The majority of these masses are benign but many can be malignant, so they need to be treated with aggressive surgical management. METHODS: In this case series, eight patients with retrorectal masses of various etiologies are described, with particular emphasis on diagnosis and surgical treatment. RESULTS: In our series, we noted one duplication cyst, one tailgut cyst, one epidermoid cyst, one teratoma, one gastrointestinal stromal tumor, one epithelial malignant tumor, one inflammatory mass, and one retrorectal mass of as yet unknown origin. In three patients, complete excision via posterior sagittal approach was performed, one underwent abdominoperineal en-block proctectomy, two were only biopsied for neoadjuvant therapy, and two are waiting for operation. CONCLUSIONS: Surgery is the main treatment of choice, and the surgical strategy should be decided according to the localization and nature of the retrorectal mass. PMID- 18386240 TI - The buried bumper syndrome: the usefulness of retrieval PEG tubes in its management. AB - Percutaneous endoscopic gastrostomy is a safe and easy method and carries a low mortality and complication rate. The buried bumper syndrome is a rare and late complication of percutaneous endoscopic gastrostomy tube placement. An 80-year old man with bilateral basal ganglia bleeding was unable to swallow safely and required tube feeding. A Flexiflo Inverta percutaneous endoscopic gastrostomy tube was successfully inserted by pull technique. One year later, he was readmitted to our clinic because of nonfunctioning tube and peristomal cellulites. Endoscopy demonstrated dimpling of the gastric mucosa on the anterior wall of the stomach. Abdominal computed tomography revealed the bumper to be buried in the abdominal wall. The tube was removed by external traction, without any abdominal incision, and a different site was used for the insertion of a new percutaneous endoscopic gastrostomy tube. No further problems were encountered over the follow-up period of nine months. As a result, the Flexiflo Inverta percutaneous endoscopic gastrostomy tubes with externally removable internal bumpers were found useful in the treatment of buried bumper syndrome, and the buried bumper was easily removed by external traction without any endoscopic or surgical methods. PMID- 18386241 TI - Secondary aortoenteric fistula in Behcet's disease. AB - Vascular manifestations of Behcet's disease include venous and arterial occlusions, arterial aneurysm and pseudo-aneurysm formation. The main problem of the surgical treatment of vascular lesions in Behcet's disease is the high incidence of complications such as recurrent aneurysms, thrombosis and fistulization to the adjacent organs. Here we present a case of Behcet's disease with multiple complications after aortic reconstructive surgery, including perigraft infection, abscess distal to the graft, occlusion of arteries of the lower extremities, aortoenteric fistula and distal anastomotic site aneurysm rupture. PMID- 18386242 TI - Eosinophilic colitis as an unusual cause of severe bloody diarrhea. AB - Eosinophilic gastrointestinal diseases may affect the colon; however, isolated colonic involvement is very rare. Diagnosis of eosinophilic colitis requires clinical suspicion and it should be differentiated from other disorders causing eosinophil accumulation in the colon tissue. Herein we present a 46-year-old female admitted to the hospital with the complaints of bloody diarrhea (25-30 times a day), fever, abdominal pain and weight loss. Eosinophilic colitis was diagnosed and she was treated with steroid successfully. PMID- 18386243 TI - A rare cause of hemosuccus pancreaticus: primary splenic artery aneurysm ruptured into pancreatic serous cystadenoma. AB - Hemosuccus pancreaticus is a rare clinical condition defined as bleeding into the pancreatic duct from a peripancreatic artery. We present here a 57-year-old woman admitted to our clinic with abdominal pain, tar-colored stool and confusion. Further investigations were done because of severe anemia. Abdominal computerized tomography revealed intraabdominal hematoma. Laparotomy was performed, which confirmed that intraabdominal haemorrhagia had occurred with the rupture of a splenic artery aneurysm into a pancreatic serous cystadenoma, which ruptured into the abdomen because of high pressure. This is an interesting case diagnosed with multidisciplinary approaches. PMID- 18386244 TI - A rare cause of ascites: Familial Mediterranean fever. AB - Familial Mediterranean fever is an autosomal recessive disorder characterized by sporadic, paroxysmal attacks of fever and serosal inflammation. In Familial Mediterranean fever, peritoneal effusion during abdominal attacks is usually mild, is not detected by clinical evaluation, and disappears during clinical remission. Chronic ascites has rarely been described in patients with Familial Mediterranean fever. Genetic analysis is highly specific and sensitive for diagnosis of Familial Mediterranean fever. All of the four cases discussed in our study had no benign or malignant pathology that could explain the ascites. They had suffered from repetitive periods of fever and ascites since childhood. Genetic analysis of these four cases revealed that one was M694V/M694V homozygote, one was M694V/? heterozygote, and the other two were M694V/V726A compound heterozygote. Ascites regressed with colchicine therapy. Since Familial Mediterranean fever is common our country, it should be kept in mind in the differential diagnosis in patients with ascites of unknown etiology. PMID- 18386245 TI - Gastrosplenic fistula due to splenic large cell lymphoma diagnosed by percutaneous drainage before surgical treatment. PMID- 18386246 TI - Isolated cecal necrosis mimicking acute appendicitis. PMID- 18386247 TI - Granular cell tumor of colon. PMID- 18386248 TI - An unusual primary metastasis of colorectal carcinoma to the forearm. PMID- 18386249 TI - A case of severe cholestatic jaundice associated with Graves' disease. PMID- 18386250 TI - Expression of tropane alkaloids in the hairy root culture of Atropa acuminata substantiated by DART mass spectrometric technique. AB - Agrobacterium rhizogenes-mediated 'hairy root' cultures were established in Atropa acuminata. The chemical profiling of the hairy roots was carried out by a new mass spectrometric technique, direct analysis in real time (DART). The intact hairy roots were directly analyzed by holding them in the gap between the DART ion source and mass spectrometer. Two alkaloids, atropine and scopolamine, were characterized. The structural confirmation of the two alkaloids was made through their accurate molecular formula determinations. This is the first report of establishing hairy roots in A. acuminata as well as application of the DART technique for the chemical profiling of its hairy roots. PMID- 18386251 TI - Biphasic effects of Morus alba leaves green tea extract on mice in chronic forced swimming model. AB - In this study, the effects of an aqueous extract of Morus alba leaves green tea (ME) on mouse behaviors (depression, anxiety, climbing activity and thermal response), muscle coordination and muscle strength were studied. Male IRC mice received a single intraperitoneal injection of either the ME, desipramine or diazepam. Thirty minutes after injection, the mice were tested in all experimental models. A significant antidepressant-like effect could be detected in the animals receiving either 100 or 200 mg/kg ME. The effect of 200 mg/kg ME in decreasing the immobility time was comparable to 10 mg/kg desipramine. With higher dose (1000 mg/kg), a significant increase in immobility time could be observed. In the elevated plus maze, no increase in time in the open arm could be observed in mice treated with ME at either 100 or 200 mg/kg. However, high doses of ME (500 or 1000 mg/kg) decreased both time in the open arm and the number of entries in the maze. No change in thermal response could be seen in mice treated with ME at doses up to 500 mg/kg, however, at 1000 mg/kg, the response time to heat was increased significantly. The ME at either 500 or 1000 mg/kg also decreased muscle coordination, strength and climbing activity significantly when compared with the control. This study suggests that ME possesses an antidepressant- without an anxiolytic-like effect, however, at high doses, the extract might show the sedative effect and alter other functions such as muscle strength, animal activity in the maze and pain response. PMID- 18386252 TI - Inhibitory effects of Angelicae Gigantis Radix on osteoclast formation. AB - Angelicae Gigantis Radix (AGR) is one of the most widely used herbal medications. AGR is the dried root of Angelica gigas Nakai (Umbelliferae), which is known as Korean angelica. This study investigated the effects of AGR on osteoclast formation using primary bone marrow cells. TNF-alpha treatment increased tartrate resistant acid phosphatase (Trap) positive cells and Trap activity in bone marrow cells. However, AGR significantly decreased both TNF-alpha-induced Trap positive cells and Trap activity. RT-PCR analyses revealed that AGR decreased mRNA levels of Trap and matrix metalloproteinase-9 in TNF-alpha-treated bone marrow cells. In addition, AGR decreased TNF-alpha-induced activation of NF-kappaB. These results suggest that AGR has an inhibitory effect on the formation of osteoclasts and its effect is partially related to the NF-kappaB pathway. PMID- 18386253 TI - Long term exposure effect of a unique metabolic nutrition system containing a diverse group of phytochemicals on serum chemistry and genomic and non-genomic changes in the liver of female B6C3F1 mice. AB - The nutritional value and therapeutic benefits coupled with presumed safety have heightened interest in the use of custom designed dietary supplements. Their use has increased substantially since the passage of the 1994 Dietary Supplement Health Education Act. However, few well-controlled studies have been conducted to assess the safety and potential adverse effects of dietary supplements. MNSO (MNS Orange-AdvoCare) is a unique combination of vitamins, minerals, omega-3 fatty acids and herbal extracts designed to provide a strong foundation of nutritional support, enhance thermogenesis, heighten energy levels and improve immune status. This investigation was designed to explore the safety and toxic effects, if any, of 12 months of continuous exposure to the ephedra and caffeine containing MNSO on serum chemistry and histopathology of seven vital target organs in female B6C3F1 mice. MNSO is enriched with extracts of citrus, Ephedra, Ginkgo, green tea and Ocimum. In this study, mice were fed control (-MNSO) or MNSO (1x-10x, 1x = daily human dose) diets. Blood was collected from all groups including the control every 4 months for serum chemistry analysis (enzyme, lipid, carbohydrate, electrolyte profiles), and liver tissue was collected for tissue biochemistry and histopathology. Multiple biochemical parameters included: (i) determination of oxidative stress via lipid peroxidation and (ii) assessment of genomic integrity via DNA fragmentation. In addition, food consumption and body weight changes were also monitored biweekly. The data showed that 12 months ingestion of 10x-MNSO did not significantly influence organ histopathology, alter the normal serum chemistry profile or any of the crucial tissue biochemistry. MNSO-exposed animals were more active, consumed more food and were relatively leaner compared with the controls. This study indicates that a caffeine and ephedra containing metabolic nutrition system is non-toxic and non-hepatotoxic in mice at up to 10x the human consumption dose of ephedra. PMID- 18386254 TI - Antioxidant and antiinflammatory activities of Sideritis perfoliata subsp. perfoliata (Lamiaceae). AB - Sideritis perfoliata L. subsp. perfoliata is a plant widely used in folk medicine in Greece since antiquity because of its antiinflammatory, antirheumatic, antiulcer, digestive and vasoprotective properties. Phytochemical investigations of the polar extracts afforded four flavonoid glycosides, four phenylpropanoic glycosides, caffeic acid and one iridoid, ajugoside. Reactive oxygen species (ROS) are implicated in the aetiology of several inflammatory processes. In the present study polar fractions and isolated compounds from S. perfoliata subsp. perfoliata were evaluated for their antioxidant activity using DPPH spectrophotometric and TBA lipid peroxidation assays, as well as for their antiinflammatory activity using the soybean lipoxygenase bioassay. All extracts and isolated compounds showed significant antioxidant and inhibitory activity against soybean lipoxygenase. These findings give support to the ethnopharmacological use of the plant in the treatment of several inflammatory ailments. PMID- 18386255 TI - Treatment of osteoarthritis with Pycnogenol. The SVOS (San Valentino Osteo arthrosis Study). Evaluation of signs, symptoms, physical performance and vascular aspects. AB - The aim of this double-blind, placebo-controlled study was to evaluate the efficacy of 100 mg Pycnogenol daily (oral capsules) in a 3 month study in patients with osteoarthritis (OA). OA symptoms were evaluated by WOMAC scores, mobility by recording their walking performance (treadmill). Treatment (77 patients) and placebo group (79) were comparable for age, sex distribution, WOMAC scores, walking distances and use of antiinflammatory drugs. The global WOMAC score decreased by 56% (p < 0.05) in the treatment group versus 9.6% in the placebo group. Walking distance in the treadmill test was prolonged from 68 m at the start to 198 m after 3 months treatment (p < 0.05), under placebo, from 65 m to 88 m (NS). The use of drugs decreased by 58% in the treatment group (p < 0.05) versus 1% under placebo. Gastrointestinal complications decreased by 63% in the treatment group, but only 3% under placebo. Overall, treatment costs were reduced significantly compared with placebo. Foot edema was present in 76% of the patients of the treatment group at inclusion and in 79% of the controls. After 3 months edema decreased in 79% of Pycnogenol patients (p < 0.05) vs 1% in controls. In conclusion, Pycnogenol offers an option for reduction of treatment costs and side effects by sparing antiinflammatory drugs. PMID- 18386256 TI - Antioxidative activity of geranium (Pelargonium inquinans Ait) and its active component, 1,2,3,4,6-penta-O-galloyl-beta-D-glucose. AB - The antioxidative activity of geranium (Pelargonium inquinans Ait) and its active component was investigated under in vitro and cellular oxidative stress models. The MeOH extract, and n-hexane, EtOAc, BuOH and H(2)O fractions from geranium showed strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effects and protective potential from oxidative damage by the radical generator, 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) in renal epithelial LLC-PK(1) cells. In particular, the EtOAc fraction exerted the strongest antioxidative potential in not only the in vitro but also in the cellular system. It suggests that the antioxidative activity of geranium is attributed mainly to components from the EtOAc fraction. Furthermore, the active component, 1,2,3,4,6-penta-O-galloyl-beta d-glucose (PGG), was identified from the EtOAc fraction by the bioassay-linked fractionation method. It displayed a potent antioxidant effect against the DPPH radical, showing an IC(50) value of 1.14 microg/mL. Moreover, the compound recovered the cell viability declined by AAPH treatment significantly and dose dependently, implying a protective role against cellular oxidative damage. The present study suggests that geranium has an excellent antioxidative potential and that PGG from geranium is considered to be the active component with an antioxidative effect. PMID- 18386257 TI - Brucea javanica fruit induces cytotoxicity and apoptosis in pancreatic adenocarcinoma cell lines. AB - Brucea javanica fruit is thought to have anticancer properties in Chinese medicine and its extract has been shown to possess antiproliferative and pro apoptotic activities on human carcinoma cells. In the present study we demonstrated for the first time that Fructus Bruceae extract exhibited cytotoxic effects on the three pancreatic adenocarcinoma cell lines, PANC-1, SW1990 and CAPAN-1; the effects were comparable to those exhibited by camptothecin in our culture system. In addition, Fructus Bruceae extract induced fragmentation of genomic DNA, as evidenced by Hoechst staining and the cell death detection ELISA(PLUS) assay. Western blot analysis further showed down-regulation of pro caspase 3 protein expression, indicating that the observed cytotoxic effects of the extract were associated with induction of apoptosis. These findings are not only significant in the development of traditional Chinese medicine as an alternative treatment for pancreatic cancer, but also in the elucidation of the potential mechanism(s) of Fructus Bruceae extract in cancer therapy. PMID- 18386258 TI - Antifungal activity of Heterothalamus alienus metabolites. AB - The chemical study of Heterothalamus alienus gave rutin, spathulenol (1), (1R,7S) germacra-4(15),5,10(14)-trien-1beta-ol (2), sakuranetin (3), padmatin 3-acetate (4), (2R,3R)-dihydroquercetin-7,3',4'-trimethyl ether (5), (2R,3R) dihydroquercetin-7,4'-dimethyl ether (6), (2R,3R)-3-acetoxy-5,7,4' trihydroxyflavanone (7), as the main components of an antifungal extract of the aerial parts of the plant. Compound 2 showed moderate activity, with Epidermophyton floccosum being the most susceptible species (MIC = 100 microg/mL); compound 3 showed the best antifungal behavior having a broad spectrum of action and the lowest MICs. This flavanone along with flavanolol 5 showed very good activity against standardized (MIC = 31.2 microg/mL) as well as clinical isolates of Trichophyton rubrum and T. mentagrophytes (MIC ranges 31.2 62.5 microg/mL and 31.2-125 microg/mL, respectively) and demonstrated not only fungistatic but also fungicide properties. Flavanolol 6 was active against all the dermatophytes tested with MICs of 62.5-250 microg/mL. Rutin, spathulenol (1) and the 3-acetylated flavanones 4 and 7 were inactive or marginally active against the fungal panel. PMID- 18386259 TI - Liquorice-induced hypokalaemia and water retention in the absence of hypertension. AB - This is a case report of a woman who showed headache, weakness, upper-limb edema and a generalized convulsive seizure after chronic ingestion of liquorice. She was taking oral contraceptives which can predispose to liquorice toxicity. Plasma potassium, aldosterone, renin activity and albumin were below the normal level. The abdominal echography and computerized tomography scan demonstrated a perihepatic and perisplenic thin liquid layer with liquid collection in the pelvis. The bioelectrical impedance suggested a hyperhydration state. After stopping the liquorice, the laboratory and bioelectrical values normalized and clinical upper-limb edema and the liquid in the abdomen disappeared in a few days. PMID- 18386261 TI - Microarray-based detection of dye-labeled DNA by SERRS using particles formed by enzymatic silver deposition. AB - The growing interest in DNA diagnostics is addressed today by microarrays with fluoresence detection. In our approach, we utilize spatially defined arrays of short oligonucleotides on a modified glass surface. Surface enhanced resonance Raman scattering (SERRS) is used to obtain molecularly specific spectra of the Raman-active dye-labeled DNA. Nanoparticles produced by enzymatic silver deposition are used as SERS-active substrate. They grow directly on the modified oligonucleotides and only in the spatially defined areas on the chip. Furthermore, they potentially offer several advantages for SERS detection. The nanoparticles are characterized and their ability for use as SERS- and SERRS active substrate is estimated. Three different Raman-active dyes are investigated for their potential for involvement in sequence specific DNA analysis. PMID- 18386260 TI - The potential role of purine-rich element binding protein (PUR) alpha as a novel treatment target for hormone-refractory prostate cancer. AB - BACKGROUND: Hormonal therapy for advanced prostate cancer is typically effective at first, but almost all men suffer refractory disease which often is life threatening. The nuclear matrix comprises not only of the structural elements of the nucleus, but is associated with many components of the molecular machinery. Our aim is to find novel targets for the treatment of hormone-refractory prostate cancer (HRPC) by focusing on the composition of the nuclear matrix proteins (NMPs). METHODS: LN96 cells were established at our Institution after long-term culturing of LNCaP cells under androgen deprived conditions. The composition of NMPs of LNCaP cells and LN96 cells were analyzed by two-dimensional (2D) electrophoresis and spots differentially expressed were investigated by mass spectrometry for identification. Among the spots identified, we analyzed the potential functional role of the identified proteins in prostate cancer cells by establishing stable overexpressed cells. RESULTS: We found that purine-rich element binding protein (PUR)alpha was significantly repressed not only in NMPs but also in total protein and mRNA levels of LN96 cells in comparison to LNCaP cells under the same steroid deprived conditions. Moreover, PURalpha was decreased in its expression both at the protein and mRNA levels in the androgen independent prostate cancer cell lines, PC3 and DU145 in comparison to LNCaP cells. Stably overexpressing PURalpha in PC3 and DU145 cells negatively regulates cell proliferation, resulting in decreases in PCNA expression. CONCLUSION: Further dissection of the role of PURalpha in cell growth regulation may reveal a novel target for HRPC. PMID- 18386262 TI - Electron-self-exchange kinetics of the cyclooctatetraene/cyclooctatetraene radical-anion couple. AB - Rate constants k(hom) and k(het) are reported for the homogeneous electron-self exchange and the heterogeneous electrochemical electron-transfer reactions, respectively, of the cyclooctatetraene/cyclooctatetraene(-) (COT/COT(.-)) redox couple. In acetonitrile, the values k(hom) (298 K)=(5+/-3) x 10(5) M(-1) s(-1) and k(het) (295 K)=8 x 10(-3) cm s(-1) are found, whereas slightly faster rates are obtained in dimethylformamide, namely, k(hom) (298 K)=(1.6+/-0.6) x 10(6) M( 1) s(-1) and k(het) (295 K)=2 x 10(-2) cm s(-1). The k(hom) rates are obtained from electron spin resonance (ESR) line broadening whereas the k(het) rates are measured at a mercurized Pt electrode by using Nicolson's method. The slowness of both electron-transfer reactions is caused by the high inner-sphere reorganization energy that results from the inevitable conformational change that takes place upon going from the tub-like COT molecule to the planar COT(.-) anion. The rates are well-understood in terms of Marcus theory, including an additional medium inner-sphere mode which is responsible for the flattening of COT. PMID- 18386263 TI - The Cu7Sc cluster is a stable sigma-aromatic seven-membered ring. AB - Density functional theory calculations demonstrate that the global minimum of the Cu(7)Sc potential energy surface is a seven-membered ring of copper atoms with scandium in its center, yielding a planar D(7) (h) structure. Nucleus-independent chemical shifts [NICS(1)(zz) and NICS(2)(zz)] show that this cluster has aromatic character, which is consistent with the number of 4s electrons of copper and scandium plus the 3d electrons of scandium satisfying Huckel's rule. According to a canonical MO decomposition of NICS(1)(zz) and NICS(2)(zz), the MOs consisting of the 4s atomic orbitals are mainly responsible for the aromatic behavior of the cluster. The electron localizability indicator (ELI-D) and its canonical MO decomposition (partial ELI-D) suggest that a localized basin is formed in Cu(7)Sc by the copper atoms whereas the two circular localized domains are situated below and above the ring. The planar Cu(7)Sc cluster can thus be considered as a sigma aromatic species. These findings agree with the phenomenological shell model. PMID- 18386264 TI - Orientation ordering of nanoparticle Ag/Co cores controlled by electric and magnetic fields. AB - The effect of electric and magnetic fields on the sandwich structure Pt/hydrogenated amorphous silicon (a-Si:H)/stearic acid monolayer/Langmuir Blodgett film of Ag/Co nanoparticles encapsulated in an organic envelope is studied. This structure is used as a working electrode in an electrochemical cell filled with NaCl solution (1 mM) and equipped with an Ag/AgCl reference electrode. Reversible changes in voltammograms are observed due to treatments (negative or positive bias voltage and simultaneous laser irradiation) applied to the designed structure before measurements. As an explanation of the observed phenomena we suggest that both the Co-up and Ag-up (on the a-Si:H surface) orientation orderings of nanoparticle Ag/Co cores are repeatedly reached. The role of the photovoltaic material (a-Si:H) in the observed behavior is explained. Voltammetric measurements with an applied magnetic field support our idea about the orientation ordering of nanoparticle cores. PMID- 18386265 TI - Determination of NMR lineshape anisotropy of guest molecules within inclusion complexes from molecular dynamics simulations. AB - Nonspherical cages in inclusion compounds can result in non-uniform motion of guest species in these cages and anisotropic lineshapes in NMR spectra of the guest. Herein, we develop a methodology to calculate lineshape anisotropy of guest species in cages based on molecular dynamics simulations of the inclusion compound. The methodology is valid for guest atoms with spin 1/2 nuclei and does not depend on the temperature and type of inclusion compound or guest species studied. As an example, the nonspherical shape of the structure I (sI) clathrate hydrate large cages leads to preferential alignment of linear CO(2) molecules in directions parallel to the two hexagonal faces of the cages. The angular distribution of the CO(2) guests in terms of a polar angle theta and azimuth angle phi and small amplitude vibrational motions in the large cage are characterized by molecular dynamics simulations at different temperatures in the stability range of the CO(2) sI clathrate. The experimental (13)C NMR lineshapes of CO(2) guests in the large cages show a reversal of the skew between the low temperature (77 K) and the high temperature (238 K) limits of the stability of the clathrate. We determine the angular distributions of the guests in the cages by classical MD simulations of the sI clathrate and calculate the (13)C NMR lineshapes over a range of temperatures. Good agreement between experimental lineshapes and calculated lineshapes is obtained. No assumptions regarding the nature of the guest motions in the cages are required. PMID- 18386266 TI - Exceptionally long-lived luminescence emitted from Tb(III) ion caged in an Ag(I) Tb(III)-thiacalix[4]arene supramolecular complex in water. AB - The compositions and photophysical properties of luminescent ternary complexes of thiacalix[4]arene-p-sulfonate (TCAS), Tb(III), and Ag(I) ions were determined. At pH 6, Ag(I) (2)Tb(III) (2)TCAS(2) formed. Moreover, at pH 10, in the presence of a 20-fold excess of Ag(I) and a 50-fold excess of TCAS with respect to Tb(III), Ag(I) (2)Tb(III)TCAS(2) formed as the main luminescent species. The structure of these complexes was proposed: two TCAS ligands are linked by two S-Ag(I)-S linkages to adopt a double-cone supramolecular structure. Furthermore, each Tb(III) ion in the former complex accepts O(-), S, O(-) donation, whereas in the latter, the Tb(III) center accepts eightfold O(-) donation. The luminescence quantum yield (Phi) of Ag(I) (2)Tb(III) (2)TCAS(2) (0.16) was almost equal to that of Tb(III)TCAS, but the luminescence lifetime tau of the former (=1.09 ms) was larger than that of the latter. For Ag(I) (2)Tb(III)TCAS(2), the yield Phi (=0.11) was small, which is attributed to the low efficiency of photosensitization (eta=0.11). However, the tau value (4.61 ms) was exceptionally large and almost equal to the natural luminescence lifetime of Tb(III) (4.7 ms), which is due to the absence of coordinating water molecules (q=0.1). This is compatible with the proposed structure in which the Tb(III) ion is shielded by a supramolecular cage that expels coordinated water molecules responsible for luminescence quenching. PMID- 18386267 TI - Mesoporous titania thin film with highly ordered and fully accessible vertical pores and crystalline walls. AB - We report the preparation of mesoporous titania thin films with the R$?bar 3$m pore structure derived from the Im$?bar 3$m self-assembled ordering of the titania species and an EO(106)PO(70)EO(106) triblock copolymer. The films were spin-cast and then aged at 18 degrees C at a relative humidity of 70 %, which led to the orientation of the Im$?bar 3$m structure with the [111] direction perpendicular to the substrates. The [111] body-diagonal channels became vertical channels upon calcination at 400 degrees C, thus leading to thin films with vertical channels. The pores are ordered over a large area of up to 1 mum(2). The titania films can be formed on various types of substrates. By using a titania film formed on a Pt-coated Si wafer as a template, we produced by an electrochemical-deposition technique arrays of gold nanowires, whose morphology suggests that most of the pores of the titania thin films are accessible. The pore structure of vertical channels is stable up to 600 degrees C, at which temperature the wall materials crystallize into anatase. PMID- 18386268 TI - Van der waals and polar intermolecular contact distances: quantifying supramolecular synthons. AB - Crystal structures are viewed as being determined by ranges and constraints on interatomic contact distances between neighboring molecules. These distances are considered to arise from environment-dependent atomic sizes, that is, larger sizes for isotropic, van der Waals type contacts and smaller sizes for more polar, possibly ionic contacts. Although the idea of different, or anisotropic, radii for atoms is not new, we developed a method of obtaining atomic sizes that is based on a theoretical framework. Using different atomic sizes for the same atom in different environments, we were able to rationalize some structural observations and anomalies. For example, benzene with the Pbca structure may be described in terms of two types of CH interactions: a longer contact largely of the van der Waals type, and a shorter, structure-determining type (C(delta )H(delta+)), which we term "n-polar". Our approach is illustrated with three examples: 1) the equivalence in crystal packing of fluorobenzene, benzonitrile, pyridine N-oxide, and pyridine/HF 1:1 molecular complex, all of which take the not-so-common tetragonal P4(1)2(1)2 space group and are practically isomorphous; 2) the similarity of the Pa3 acetylene and Pbca benzene crystal structures; and 3) the equivalence between an increase in pressure and an increase in the "n polar" contacts in Pbca benzene; in other words, the equivalence between hydrostatic pressure and chemical pressure. In the context of crystal engineering, we describe a method whereby the topological information conveyed in a supramolecular synthon is recast in a more quantitative manner. A particular synthon, and in turn the crystal structure to which it leads, is viable within small ranges of distances of its constituent atoms, and these distances are determined by chemical factors. PMID- 18386269 TI - Rhodium-catalyzed oxidative coupling between salicylaldehydes and internal alkynes with C-H bond cleavage to produce 2,3-disubstituted chromones. AB - A direct oxidative coupling of salicylaldehydes with internal alkynes proceeds efficiently with cleavage of the aldehyde C--H bond to produce the corresponding chromone derivatives. A rhodium catalyst in combination with a cyclopentadiene ligand and a copper oxidant promote this straightforward annulation reaction. Solid-state luminescence was observed for certain chromone products. PMID- 18386270 TI - Methylgallium as a terminal ligand in [(Cp*Ga)4Rh(GaCH3)]+. PMID- 18386271 TI - Asymmetric epoxidation of olefins with hydrogen peroxide - catalysis by an aspartate-containing tripeptide. PMID- 18386272 TI - The enantioselective, Bronsted acid catalyzed, vinylogous Mannich reaction. PMID- 18386273 TI - Carbon nanotube-polymer composite for light-driven microthermal control. PMID- 18386274 TI - Near-infrared dichromic fluorescent carbocyanine molecules. PMID- 18386275 TI - Photolabile glutamate protecting group with high one- and two-photon uncaging efficiencies. AB - A pi-extended [2-(2-nitrophenyl)propoxy]carbonyl (NPPOC) derivative has been prepared as an efficient UV and near-IR photolabile protecting group for glutamate. This glutamate cage compound exhibits efficient photorelease upon one photon excitation (epsilonPhi=990 M(-1) cm(-1) at 315 nm). In addition, it also shows efficient photorelease in activation of glutamate receptors in electrophysiological recordings. Combined with a high two-photon uncaging cross section (deltaPhi=0.45 GM at 800 nm), its overall properties make this new cage-3 (2-propyl)-4'-methoxy-4-nitrobiphenyl (PMNB)-for glutamate a very promising tool for two-photon neuronal studies. PMID- 18386277 TI - Efficient N-terminal glycoconjugation of proteins by the N-end rule. PMID- 18386276 TI - Fluorescent agonists for the Torpedo nicotinic acetylcholine receptor. AB - We have synthesized a series of fluorescent acylcholine derivatives carrying different linkers that vary in length and structure and connect the acylcholine unit to the environment-sensitive fluorophores 7-(diethylamino)coumarin-3 carbonyl (DEAC) or N-(7-nitrobenz-2-oxa-1,3-diazol-yl) (NBD). The pharmacological properties of the fluorescent analogues were investigated on heterologously expressed nicotinic acetylcholine receptor (nAChR) from Torpedo californica and on oocytes transplanted with nAChR-rich Torpedo marmorata membranes. Agonist action strongly depends on the length and the structure of the linker. One particular analogue, DEAC-Gly-C6-choline, showed partial agonist behavior with about half of the maximum response of acetylcholine, which is at least 20 times higher than those observed with previously described fluorescent dansyl- and NBD acylcholine analogues. Binding of DEAC-Gly-C6-choline to Torpedo nAChR induces a strong enhancement of fluorescence intensity. Association and displacement kinetic experiments revealed dissociation constants of 0.5 nM for the alphadelta binding site and 15.0 nM for the alphagamma-binding site. Both the pharmacological and the spectroscopic properties of this agonist show great promise for characterizing the allosteric mechanism behind the function of the Torpedo nAChR, as well as for drug-screening studies. PMID- 18386278 TI - Identification of trinucleotide repeat ligands with a FRET melting assay. AB - DNA hairpin structures formed within a repeated tract might be a causative factor for triplet expansion observed in several debilitating diseases. We have designed and used a fluorescence resonance energy transfer (FRET) melting assay to screen for ligands that bind specifically to the CNG triplet repeats. Using this assay, we screened a panel of 33 chemicals that were previously designed to bind DNA or RNA secondary structures. Remarkably, we found that macrocyclic compounds, such as acridine dimers and trimers, exhibit interesting affinities and specificities for this motif. PMID- 18386279 TI - Species barrier of HIV-1 and its jumping by virus engineering. AB - Monkey infection models are absolutely necessary for studies of human immunodeficiency virus type 1 (HIV-1) pathogenesis and of developing drugs/vaccines against HIV-1. In addition, currently unknown roles of its accessory proteins for in vivo replication await elucidation by experimental approaches. Due to the fact that HIV-1 is tropic only for chimpanzees and humans, studies of this line have been impeded for a long time, although various investigations have been carried out utilising genetically related SIV and SIV/HIV chimeric virus (SHIV) as pathogens. Recent findings of anti-HIV-1 innate factors such as tripartite motif protein 5alpha (TRIM5alpha) and APOBEC3G/F prompted us to re-initiate an old and vital research project which would, as a result, confer the capability to overcome the species barrier on the HIV-1. We currently have obtained, by virus engineering through genetic manipulation and adaptation, some new and promising HIV-1 clones for in vivo studies in macaque monkeys as mentioned above. In this review, we summarise the past, present and future of HIV-1/SIV chimeric viruses with special reference to relevant basic HIV 1/SIV studies. PMID- 18386280 TI - Symmetrically tetrasubstituted [2.2]paracyclophanes: their systematization and regioselective synthesis of several types of bis-bifunctional derivatives by double electrophilic substitution. AB - The possible number of chiral and achiral tetrasubstituted [2.2]paracyclophanes possessing different types of symmetry (C(2), C(i), C(s), C(2v), C(2h)) is evaluated and a unified independent trivial naming descriptor system is introduced. The reactivity and regioselectivity of the electrophilic substitution of the chiral pseudo-meta- and achiral pseudo-para-disubstituted [2.2]paracyclophanes are investigated in an approach suggested to be general for the synthesis of bis-bifunctional [2.2]paracyclophanes. The mono- and diacylation of chiral pseudo-meta-dihydroxy[2.2]paracyclophane 14 with acetylchloride occur ortho-regioselectively to produce tri- 22, 23 and symmetrically 21 tetrasubstituted acyl derivatives. The same reaction with benzoylchloride is neither regio-, nor chemoselective, and gives rise to a mixture of ortho-/para-, mono-/diacylated compounds 27-31. The double acylation of pseudo-meta dimethoxy[2.2]paracyclophane 18 is completely para-regioselective. Electrophilic substitution of pseudo-meta-bis(methoxycarbonyl)[2.2]paracyclophane 20 regioselectively generates the pseudo-gem-substitution pattern. Formylation of this substrate produces the monocarbonyl derivatives 35 only, whereas the Fe catalyzed bromination may be directed towards mono- 36 or disubstitution 37 products chemoselectively by varying the reactions conditions. The diacylation and dibromination reactions of the respective achiral diphenol 12 and bis(methoxycarbonyl) 40 derivatives of the pseudo-para-structure retain regioselectivities which are characteristic for their pseudo-meta-regioisomers. Imino ligands 26, 25, and 39, which were obtained from monoacyl- 22 and diacyldihydroxy[2.2]paracyclophanes 21, 38, are tested as chiral ligands in stereoselective Et(2)Zn addition to benzaldehyde producing 1-phenylpropanol with ee values up to 76 %. PMID- 18386281 TI - Polyhedral structures with three-, four-, and five fold symmetry in metal centered ten-vertex germanium clusters. AB - Studies using density functional theory (DFT) at the hybrid B3LYP level indicate that the relative energies of structures with three-fold, four-fold, and five fold symmetry for centered 10-vertex bare germanium clusters of the general type M@Ge(10) (z) depend on the central metal atom M and the skeletal electron count. For M@Ge(10) clusters with 20 skeletal electrons the DFT results agree with experimental data on the isoelectronic centered 10-vertex bare metal clusters. Thus the lowest energy structure for Ni@Ge(10), isoelectronic with the known Ni@In(10) (10-), is a C(3v) polyhedron derived from the tetracapped trigonal prism. However, Zn@Ge(10) (2+) is isoelectronic with the known cluster Zn@In(10) (8-), which has the lowest energy structure, a D(4d) bicapped square antiprism. For the clusters Ni@Ge(10) (2-), Cu@Ge(10) (-), and Zn@Ge(10) that have 22 skeletal electrons the lowest energy structures are the D(4d) bicapped square antiprism predicted by the Wade-Mingos rules. For the clusters Ni@Ge(10) (4-), Cu@Ge(10) (3-), and Zn@Ge(10) (2-) that have 24 skeletal electrons the lowest energy structures are C(3v) polyhedra with 10 triangular faces and 3 quadrilateral faces derived from a tetracapped trigonal prism by extreme lengthening of the edges of the capped triangular face of the underlying trigonal prism. For the clusters Cu@Ge(10) (5-) and Zn@Ge(10) (4-) that have 26 skeletal electrons the lowest energy structures are the D(5d) pentagonal antiprisms predicted by the Wade-Mingos rules and the C(3v) tetracapped trigonal prism as a somewhat higher energy structure. However, for the isoelectronic Ni@Ge(10) (6-) the relative energies of these two structure types are reversed so that the C(3v) tetracapped trigonal prism becomes the global minimum. The effects of electron count on the geometries of the D(5d) pentagonal prism and D(4d) bicapped square antiprism centered metal cluster structures are consistent with the bonding/antibonding characteristics of the corresponding HOMO and LUMO frontier molecular orbitals. PMID- 18386282 TI - High-relaxivity magnetic resonance imaging (MRI) contrast agent based on supramolecular assembly between a gadolinium chelate, a modified dextran, and poly-beta-cyclodextrin. AB - Nanosized contrast agents have great potential in magnetic resonance molecular imaging applications for clinical diagnosis. This study proposes new nanoparticles spontaneously formed under mild conditions and composed of a noncovalent adduct between a gadolinium complex, a polymer of beta-cyclodextrin (pbetaCD: MW 1.5 x 10(6) g mol(-1)) and a dextran grafted with alkyl chains (MD). The formation of this supramolecular nanoassembly is based upon a "lock-and-key" recognition process in which the hydrophobic alkyl chains of MD and the adamantyl moieties of macrocyclic Gd(III) chelates are included in the cavities of pbetaCD. The large number of betaCDs contained in the pbetaCD resulted in the formation of 200 nm diameter nanoparticles, each entrapping 1.8 x 10(5) molecules of a low molecular-weight Gd complex. This system, which exhibits a great relaxivity enhancement (48.4 mM(-1) s(-1), at 20 MHz and 37 degrees C) compared to the Gd(III) chelate itself (5.2 mM(-1) s(-1)), appears to be a promising strategy for the in vivo targeted delivery of Gd(III) complexes. The mechanisms of particle formation, conjugation strategies, and relaxometric characterizations in the field of contrast-enhanced magnetic resonance imaging are discussed. PMID- 18386283 TI - Four generations of water-soluble dendrimers with 9 to 243 benzoate tethers: synthesis and dendritic effects on their ion pairing with acetylcholine, benzyltriethylammonium, and dopamine in water. AB - Water-soluble benzoate-terminated dendrimers of four generations (from G0 with 9 branches to G3 with 243 branches) were synthesized and fully characterized. They form water-soluble assemblies by ion-pairing interactions with three cations of medicinal interest (acetylcoline, benzyltriethylammonium, and dopamine), which were characterized and investigated by 1H NMR spectroscopy, whereas such interactions do not provoke any significant shift of 1H NMR signals with the monomeric benzoate anion. The calculated association constants confirm that the dendritic carboxylate termini reversibly form ion pairs and aggregates. Diffusion coefficients and hydrodynamic diameters of the dendrimers, as well as changes thereof on interaction with the cations, were evaluated by DOSY experiments. The lack of increase of dendrimer size on addition of the cations and the upfield shifts of the 1H NMR signals of the cation indicate encapsulation within the hydrophobic dendrimer interiors together with probable backfolding of the benzoate termini. PMID- 18386284 TI - A highly selective and sensitive fluorescence probe for the hypochlorite anion. AB - A new rhodamine B-based fluorescent probe for the hypochlorite anion (OCl(-)) has been designed, synthesized, and characterized. The probe comprises a spectroscopic unit of rhodamine B and an OCl(-)-specific reactive moiety of dibenzoylhydrazine. The probe itself is nearly nonfluorescent because of its spirolactam structure. Upon reaction with OCl(-), however, a largely enhanced fluorescence is produced due to the opening of the spirolactam ring by the oxidation of the exocyclic hydrazide and subsequently the formation of the hydrolytic product rhodamine B. Most notably, the fluorescence-on reaction shows high sensitivity and extremely high selectivity for OCl(-) over other common ions and oxidants, which makes it possible for OCl(-) to be detected directly in their presence. In addition, the reaction mechanism has been investigated and proposed. The OCl(-) anion selectively oxidizes the hydrazo group in the probe, and forms the analogue of dibenzoyl diimide, which in turn hydrolyzes and releases the fluorophore. The reaction mechanism that is described here might be useful in developing excellent spectroscopic probes with cleavable active bonds for other species. PMID- 18386285 TI - JunD mediates androgen-induced oxidative stress in androgen dependent LNCaP human prostate cancer cells. AB - BACKGROUND: Numerous and compelling evidence shows that high level of reactive oxygen species (ROS) plays a key role in prostate cancer occurrence, recurrence and progression. The molecular mechanism of ROS overproduction in the prostate gland, however, remains mostly unknown. Unique AP-1 transcription factor JunD has been shown to inhibit cell proliferation, promote differentiation and mediate stress responses in a variety of eukaryotic cells. We previously reported that androgen-androgen receptor induced ROS production in androgen-dependent LNCaP human prostate cancer cells is associated with increased JunD level/AP-1 transcriptional activity. METHODS: LNCaP cells constitutively overexpressing a functionally inactive form of JunD (JunDDeltaTA) or stably transfected with JunD siRNA (siJunD) to suppress JunD protein expression were established. Overexpression of JunD in LNCaP cells using transient transfection method was applied to assess the induction of ROS production in LNCaP cells. DCF assay was used to measure the ROS concentrations in the transfected as well as non transfected control cells. RT-PCR and Western blot analyses were used to confirm silencing or overexpression of JunD in the transfected cells. RESULTS: In the absence of androgen, LNCaP cells transiently transfected with a JunD overexpressing vector have relatively enhanced cellular ROS levels as compared to LNCaP cells transfected with a vector control. LNCaP cells that fail to express functional JunD (JunDDeltaTA or siJunD) do not exhibit any increase in ROS production in response to androgen. CONCLUSION: Based on these data, we conclude that JunD is an essential mediator of the androgen-induced increase in ROS levels in LNCaP cells. PMID- 18386286 TI - Fibroblast growth factor 7 stimulates in vitro growth of oocytes originating from bovine early antral follicles. AB - Essential factors required for growing oocytes derived from bovine early antral follicles and their mechanisms of action are poorly understood. Fibroblast growth factor 7 (FGF7) is a member of the heparin-binding FGF family with a distinctive pattern of target-cell specificity. The effect of FGF7 on the stimulation of oocyte growth in a culture of cumulus-oocyte complexes with granulosa cells (COCGs, oocyte diameter; 90-100 microm) was investigated. The oocyte diameter of COCGs was increased significantly in the FGF7-containing medium (10 ng/ml; 117.2 +/- 3.2 microm, 50 ng/ml; 116.5 +/- 3.5 microm) compared to the control (0 ng/ml; 110.5 +/- 2.8 microm) after 16 days. However, there was no stimulatory effect of FGF7 on the proliferation of cumulus-granulosa cells. The FGF7 receptor, fibroblast growth factor receptor 2IIIb (FGFR2IIIb), was detected in cumulus granulosa cells from COCGs. Messenger RNA expression of FGFR2IIIb was induced to cumulus-granulosa cells by FGF7. The mRNA expression levels of KIT ligand (KITLG), KIT (KIT), growth differentiation factor 9 (GDF9), and bone morphogenetic protein 15 (BMP15) in the cultured COCGs were determined in FGF7 treated (10 ng/ml) cultures using real time RT-PCR analysis. The levels of KITLG and KIT, but not GDF9 and BMP15 mRNA expression were stimulated by FGF7. Furthermore, neutralizing antibody for KIT attenuated the stimulatory action of FGF7 on the oocyte growth. These results strongly suggest that FGF7 may be an important regulator for oocyte growth and its action is mediated via the KIT/KITLG signaling pathway. PMID- 18386287 TI - Regulation of cap-dependent translation initiation in the early stage porcine parthenotes. AB - The binding of mRNAs to ribosomes is mediated by the protein complex eIF4F in conjunction with eIF4B (eukaryotic initiation factor 4F and 4B). EIF4F is a three subunit complex consisting of eIF4A (RNA helicase), eIF4E (mRNA cap binding protein), and eIF4G (bridging protein). The crucial role is played by eIF4E, which directly binds the 5'-cap structure of the mRNA and facilitates the recruitment to the mRNA of other translation factors and the 40S ribosomal subunit. EIF4E binding to mRNA and to other initiation factors is regulated on several levels, including its phosphorylation on Ser-209, and association with its regulatory protein 4E-binding protein (4E-BP1). In this study we document that both the translation initiation factor eIF4E and its regulator 4E-BP1 become dephosphorylated in the early stage porcine zygotes already 8 hr post-activation. Similarly, the activities of ERK1/2 MAP and Mnk1 kinases, which are both involved in eIF4E phosphorylation, gradually decrease during this period with the timing similar to that of eIF4E dephosphorylation. The formation of an active eIF4F complex is also diminished after 9-15 hr post-activation, although substantial amounts of this complex have been detected also 24 hr post-activation (2-cell stage). The overall protein synthesis in the parthenotes decreases gradually from 12 hr post-activation reaching a minimum after 48 hr (4-cell stage). Although the translation is gradually decreasing during early preimplantation development, the eIF4F complex, which is temporarily formed, might be a premise for the translation of a small subset of mRNAs at this period of development. PMID- 18386288 TI - Protease-activated receptor-1 upregulates fibroblast growth factor 7 in stroma of benign prostatic hyperplasia. AB - BACKGROUND: Benign prostatic hyperplasia (BPH) is characterized by abnormal epithelial and stromal proliferation causing urinary obstruction. Prostate growth is regulated by a variety of growth factors secreted from the stroma, including fibroblast growth factor 7 (FGF-7), a potent epithelial-specific growth factor which is increased in hyperplastic prostate. However, the mediator(s) of FGF-7 over-expression is unclear. Protease-activated receptor-1 (PAR-1) is a G-protein coupled receptor known to induce multiple biological processes, but its effect on BPH pathogenesis is mostly unknown. The aim of this study was to investigate the role of PAR-1 as a mediator of BPH development. METHODS: PAR-1 expression was investigated in BPH and normal prostate tissues by immunohistochemistry. Prostate stromal cells were isolated from BPH specimens, cultured and immunohistochemically characterized. Cultured stromal cells were stimulated with PAR-1 agonists, and extracellular-signal regulated kinase (ERK1/2) activation and cell proliferation were examined. PAR-1 mediated FGF-7 production by cultured stromal cells was assessed by RT-PCR and immunoassays, and verified by small interfering RNA (siRNA). RESULTS: PAR-1 expression was increased in BPH stroma. In stromal cells isolated from BPH tissues, PAR-1 agonists activated ERK1/2 in a time- and concentration-dependent manner and with resultant enhanced cell proliferation. Pertussis toxin-sensitive G protein/(betagamma-subunits) phosphatidylinositol 3-kinase and protein kinase C pathways were involved in ERK1/2 phosphorylation. PAR-1 activation strikingly induced FGF-7 production from cultured stromal cells mediated predominantly via ERK1/2 signaling pathway, and PAR-1 siRNA decreased the elicited FGF-7 upregulation. CONCLUSIONS: The expression and function of PAR-1 in BPH stroma indicate PAR-1 may play important roles in BPH pathogenesis. PMID- 18386289 TI - Structural characterization and anti-angiogenic properties of prostate-specific antigen isoforms in seminal fluid. AB - BACKGROUND: The prostate produces high levels of prostate-specific antigen (PSA), which has been shown to exert anti-angiogenic properties and thus might slow down prostate tumor growth. It has been suggested that the protease activity of PSA is not needed for its anti-angiogenic function. We have previously shown that seminal fluid contains both active and inactive, internally cleaved forms of PSA. The precise structural differences between these isoforms and their function are not known. METHODS: To elucidate the structures, we purified PSA from seminal fluid and separated it by anion-exchange chromatography into six different isoforms, which were characterized by mass spectrometry. The anti-angiogenic activity of these PSA-isoforms was analyzed by human umbilical vein endothelial cell (HUVEC) tube formation assay. RESULTS: The enzymatically active PSA-isoforms had an intact peptide moiety but could be separated into three isoforms based on differences in glycosylation. The major isoform contained PSA with a biantennary carbohydrate with terminal sialic acids on both antennae. The other active isoforms showed significant carbohydrate heterogeneity, containing one or no sialic acid. The inactive isoforms were internally cleaved at several different positions, but the fragments were held together by disulphide bonds. The enzymatic activity of PSA correlated with its inhibitory effect on the endothelial cell tube formation and the inhibition was dose-dependent at physiological concentrations, whereas enzymatically inactive internally cleaved PSA-isoforms had no effect. CONCLUSIONS: Our results show that the anti angiogenic effect of PSA is based on its proteolytic activity. PMID- 18386290 TI - Detection and characterization of gamete-specific molecules in Mytilus edulis using selective antibody production. AB - The mussel Mytilus edulis can be used as model to study the molecular basis of reproductive isolation because this species maintains its species integrity, despite of hybridizing in zones of contact with the closely related species M. trossulus or M. galloprovincialis. This study uses selective antibody production by means of hybridoma technology to identify molecules which are involved in sperm function of M. edulis. Fragmented sperm were injected into mice and 25 hybridoma cell clones were established to obtain monoclonal antibodies (mAb). Five clones were identified producing mAb targeting molecules putatively involved in sperm function based on enzyme immunoassays, dot and Western blotting as well as immunostaining of tissue sections. Specific localization of these mAb targets on sperm and partly also in somatic tissue suggests that all five antibodies bind to different molecules. The targets of the mAb obtained from clone G26-AG8 were identified using mass spectrometry (nano-LC-ESI-MS/MS) as M6 and M7 lysin. These acrosomal proteins have egg vitelline lyses function and are highly similar (76%) which explains the cross reactivity of mAb G26-AG8. Furthermore, M7 lysin was recently shown to be under strong positive selection suggesting a role in interspecific reproductive isolation. This study shows that M6 and M7 lysin are not only found in the sperm acrosome but also in male somatic tissue of the mantle and the posterior adductor muscle, while being completely absent in females. The monoclonal antibody G26-AG8 described here will allow elucidating M7/M6 lysin function in somatic and gonad tissue of adult and developing animals. PMID- 18386292 TI - Clinical assessment of the cancer diagnostic value of prostatic zinc: a comprehensive needle-biopsy study. AB - BACKGROUND: The correlation between Zinc concentration in the prostate's peripheral zone to the onset or presence of malignant process needs to be evaluated in detail. METHODS: Zinc concentration was measured in approximately 1 4 mm3 segments of fresh needle-biopsy cores, with X-ray fluorescence, and correlated with the histological findings of these tissue segments. RESULTS: Local Zinc concentration is correlated with the presence of cancer (PCa); the higher the Gleason score the greater the Local Zinc depletion. The Zinc value averaged over the entire extracted tissue is specific only to Gleason score 8-9 PCa. The results refer to patients avoiding Zinc-rich supplements since those show elevated prostatic Zinc concentration in identified cancer tissue. A computer simulation analysis of randomly located 0.03-3.3 cm3 lesions, with particular Gleason score and the measured Local Zinc concentration, revealed a potential diagnostic approach definitely superior to PSA, with sensitivity to the tumor grade and with excellent detection capability for Gleason score >6. Further clinical studies have been designed, both on full prostates after radical prostatectomy as well as on biopsy cores at higher resolution, to establish the accuracy of the method for Gleason score = 6. CONCLUSIONS: The PCa diagnostic potential of Local Zinc concentration is confirmed and there is indication that the amount of Zinc depletion could be used as a measure of the Gleason score of the tumor. Local Zinc concentration mapping has the potential to improve patient selection for biopsy, biopsy site selection and local therapy (e.g., Cryotherapy, Brachytherapy) site selection. PMID- 18386291 TI - Restoration of PTEN expression alters the sensitivity of prostate cancer cells to EGFR inhibitors. AB - INTRODUCTION: Prostate cancer (CaP) progression from an androgen-dependent to an androgen-independent state is associated with overexpression of EGFR family members or activation of their downstream signaling pathways, such as PI3K-Akt and MAPK. Although there are data implicating PI3K-Akt or MAPK pathway activation with resistance to EGFR inhibitors in CaP, the potential cross-talk between these pathways in response to EGFR or MAPK inhibitors remains to be examined. METHODS: Cross-talk between PTEN and MAPK signaling and its effects on CaP cell sensitivity to EGFR or MAPK inhibitors were examined in a PTEN-null C4-2 CaP cell, pTetOn PTEN C4-2, where PTEN expression was restored conditionally. RESULTS: Expression of PTEN in C4-2 cells exposed to EGF or serum was associated with increased phospho-ERK levels compared to cells without PTEN expression. Similar hypersensitivity of MAPK signaling was observed when cells were treated with a PI3K inhibitor LY294002. This enhanced sensitivity of MAPK signaling in PTEN-expressing cells was associated with a growth stimulatory effect in response to EGF. Furthermore, EGFR inhibitors gefitinib and lapatinib abrogated hypersensitivity of MAPK signaling and cooperated with PTEN expression to inhibit cell growth in both monolayer and anchorage-independent conditions. Similar cooperative growth inhibition was observed when cells were treated with the MEK inhibitor, CI1040, in combination with PTEN expression suggesting that inhibition of MAPK signaling could mediate the cooperation of EGFR inhibitors with PTEN expression. CONCLUSIONS: Our results suggest that signaling cross-talk between the PI3K-Akt and MAPK pathways occurs in CaP cells, highlighting the potential benefit of targeting both the PI3K-Akt and MAPK pathways in CaP treatment. PMID- 18386293 TI - Prostate stem cells and benign prostatic hyperplasia. AB - Pharmacological approaches are available to medically-managed patients with symptomatic BPH before surgical intervention is required. These include daily treatment with alpha-blockers and 5-alpha-reductase inhibitors alone or in combination. These medical approaches have two major problems. First, treatments are chronic and must be taken daily. Second, there are significant financial costs and quality of life issues for such chronic treatments. Is it possible to develop effective acute therapy for symptomatic BPH without the long-term androgen deprivation-induced side effects? Two seminal but rarely cited studies of Walsh [Peters, Walsh: N Engl J Med 317:599-604, 1987] and Coffey et al. [Sufrin et al.: Invest Urol 13:418-423, 1976], combined with the growing understanding of the stem cell organization of the prostate stromal (S) and epithelial (E) compartments and their reciprocal paracrine and autocrine interactions provides the rationale for an acute approach.The Walsh study documents that: (1) androgen deprivation disrupts the reciprocal interaction between the prostate S and E thereby decreasing the weight of both compartments and (2) once BPH develops, androgen deprivation does not decrease the number of stem cell units in either the S or E compartments since subsequent androgen restoration fully restores the enlarged gland. The Coffey study documents that acute androgen deprivation sensitizes S-E interactions to radiation induced disruptions so that following radiation, androgen restoration does not induce full gland regrowth. Therefore, effective therapy for symptomatic BPH should be achievable by acute treatment with reversible androgen deprivation for a limited period followed by a single dose of conformal external beam radiation before allowing the man to recovery his normal serum testosterone. PMID- 18386294 TI - Levels of adiponectin, C-reactive protein and interleukin-1 receptor antagonist are associated with insulin sensitivity: a population-based study. AB - BACKGROUND: We evaluated the relationship of insulin sensitivity (assessed with the quantitative insulin sensitivity check index, QUICKI) to adiponectin and pro inflammatory markers, levels of high-sensitivity C-reactive protein (hs-CRP) and interleukin-1 receptor antagonist (IL-1 Ra). METHODS: Cross-sectional study. Study population (N=923, i.e 411 men and 512 women) included five different population-based age groups (born in 1942, 1947, 1952, 1957 and 1962), [mean age 46 years and mean body mass index (BMI) 26 kg/m(2)]. Study protocol included an interview and measurements of anthropometric parameters and glucose, insulin, adiponectin, hs-CRP and IL-1 Ra. RESULTS: Correlation (r) between QUICKI and adiponectin level was 0.334 [95% confidence intervals (CI), 0.275-0.392] and partial correlation adjusted for gender, BMI, smoking status, physical activity and age was 0.247 (95% CI, 0.185-0.308). There was negative correlation between QUICKI and IL-1 Ra (r= -0.385; 95% CI, -0.440 to -0.328) which remained statistically significant after the adjustment for confounding factors (r= 0.178; 95% CI, -0.240 to -0.113). Similarly, QUICKI was negatively correlated with hs-CRP (r= -0.241; 95% CI, -0.302 to -0.178), but after the adjustment it lost its statistical significance. There was a statistically significant gender difference (p=0.018) in correlation between QUICKI and IL-1 Ra levels (men: r= 0.348; 95% CI, -0.436 to - 0.261; women r= -0.500; 95% CI, -0.537 to -0.398). CONCLUSIONS: Our results show that adiponectin level and markers of low-grade inflammation are related to insulin sensitivity. Adiponectin and IL-1 Ra levels might be better markers of the risk of obesity and type 2 diabetes than hs-CRP. PMID- 18386295 TI - Sulfonated molecules that bind a partially structured species of beta2 microglobulin also influence refolding and fibrillogenesis. AB - Human beta2-microglobulin (beta2-m) is a small amyloidogenic protein responsible for dialysis-related amyloidosis, which represents a severe complication of long term hemodialysis. A therapeutic approach for this amyloidosis could be based on the stabilization of beta2-m through the binding to a small molecule, to possibly inhibit protein misfolding and amyloid fibril formation. The search of a strong ligand of this protein is extremely challenging: by using CE in affinity and refolding experiments we study the effect that previously selected sulfonated molecules have on the equilibrium between the native form and an ensemble of conformers populating the slow phase of beta2-m folding. These data are correlated with the effect that the same molecules exert on in vitro fibrillogenesis experiments. PMID- 18386296 TI - Optically active zwitterionic lambda(5)Si,lambda(5)Si'-disilicates: syntheses, crystal structures, and behavior in aqueous solution. AB - The zwitterionic lambda(5)Si,lambda(5)Si'-disilicates 1-8 were synthesized and characterized by solid-state and solution NMR spectroscopy. In addition, compounds 26 H(2)O, 32 CH(3)CN, 45/2 CH(3)CN, 6CH(3)OH, 7, and 8CH(3)OHCH(3)CN were studied by single-crystal X-ray diffraction. The optically active (Delta,Delta,R,R,R,R)-configured compounds 1-8 contain two pentacoordinate (formally negatively charged) silicon atoms and two tetracoordinate (formally positively charged) nitrogen atoms. One (ammonio)alkyl group is bound to each of the two silicon centers, and two tetradentate (R,R)-tartrato(4-) ligands bridge the silicon atoms. Although these lambda(5)Si,lambda(5)Si'-disilicates contain SiO(4)C skeletons, some of them display a remarkable stability in aqueous solution as shown by NMR spectroscopy and ESI mass spectrometry. PMID- 18386297 TI - Kinetin enhances in vitro development of parthenogenetic and nuclear transfer porcine embryos. AB - Culture conditions affect the development of mammalian embryos in vitro. Kinetin belongs to the family of N(6)-substituted adenine derivates and promotes cell division, synthesis of DNA repair enzymes, superoxide dismutase activity, and ribosomal RNA transcription. We investigated the effects of kinetin on in vitro development of parthenogenetic and nuclear transfer (NT) porcine embryos. These embryos were cultured with or without kinetin in either BSA- or polyvinyl alcohol containing medium for 7 days. mRNA expression of three developmentally important genes, HSP70, Glut-1, and poly[A] polymerase in NT embryos was analyzed. Regardless of kinetin supplementation, the proportion of blastocysts and blastocyst cells were not significantly different in parthenogenetic embryos. However, kinetin supplementation increased expansion and hatching rates in all groups. In somatic cell NT embryos, kinetin increased the proportion of embryos developed to blastocysts from 7.5% to 15.4% in medium supplemented with PVA. However, gene expression levels of HSP70, poly[A] polymerase and Glut-1 mRNA were not significantly different in NT blastocysts. The present study indicates that kinetin not only improves blastocyst expansion and cell number of parthenogenetic porcine embryos but also enhances NT porcine embryo development in a completely defined culture condition in vitro. PMID- 18386298 TI - Prediction of lymphatic invasion by peritumoral lymphatic vessel density in prostate biopsy cores. AB - BACKGROUND: Lymphatic invasion in radical prostatectomy specimens has been suggested to be an unfavorable prognostic factor in clinically localized prostate cancer. Lymphangiogenesis detected by antibodies specific for lymphatic endothelial cells has been associated with lymphatic invasion and lymph node metastasis in prostate cancer. This study was designed to examine whether lymphangiogenesis in prostate biopsy could predict lymphatic spread in radical prostatectomy specimens. METHODS: Paraffin-embedded positive biopsy cores obtained from 99 patients who underwent radical prostatectomy at our institution were immunostained with D2-40 monoclonal antibody, which specifically recognizes lymphatic endothelium. The association between lymphatic parameters in prostate biopsy and pathological parameters in radical prostatectomy specimens was analyzed. RESULTS: Peritumoral and intratumoral lymphatic (ITL) vessels were observed in 90 (90.9%) and 23 cases (23.2%). Average and maximal peritumoral lymphatic vessel density (PTLD) and the presence of ITL in positive biopsy cores were significantly associated with positive biopsy core rates (P = 0.0015 for average PTLD, P < 0.0001 for maximal PTLD, and P = 0.0038 for ITL) and lymphatic vessel invasion (P < 0.0001 for average PTLD, P < 0.0001 for maximal PTLD, and P = 0.0322 for ITL). Among preoperative parameters, the biopsy Gleason score (P = 0.0092, HR = 6.108) and average PTLD (P = 0.0034, HR = 1.860) were significant predictors of lymphatic invasion in radical prostatectomy specimens in multivariate analysis. CONCLUSIONS: PTLD in prostate biopsy specimens assessed by immunohistochemistry using D2-40 antibody could be a useful parameter for predicting lymphatic spread of clinically localized prostate cancer. PMID- 18386299 TI - Is the mu-oxo-mu-peroxodiiron intermediate of a ribonucleotide reductase biomimetic a possible oxidant of epoxidation reactions? AB - Density functional calculations on a mu-oxo-mu-peroxodiiron complex (1) with a tetrapodal ligand BPP (BPP=N,N-bis(2-pyridylmethyl)-3-aminopropionate) are presented that is a biomimetic of the active site region of ribonucleotide reductase (RNR). We have studied all low-lying electronic states and show that it has close-lying broken-shell singlet and undecaplet (S=0, 5) ground states with essentially two sextet spin iron atoms. In strongly distorted electronic systems in which the two iron atoms have different spin states, the peroxo group moves considerably out of the plane of the mu-oxodiiron group due to orbital rearrangements. The calculated absorption spectra of (1,11)1 are in good agreement with experimental studies on biomimetics and RNR enzyme systems. Moreover, vibrational shifts in the spectrum due to (18)O(2) substitution of the oxygen atoms in the peroxo group follow similar trends as experimental observations. To identify whether the mu-oxo-mu-1,2-peroxodiiron or the mu-oxo-mu 1,1-peroxodiiron complexes are able to epoxidize substrates, we studied the reactivity patterns versus propene. Generally, the reactions are stepwise via radical intermediates and proceed by two-state reactivity patterns on competing singlet and undecaplet spin state surfaces. However, both the mu-oxo-mu-1,2 peroxodiiron and mu-oxo-mu-1,1-peroxodiiron complex are sluggish oxidants with high epoxidation barriers. The epoxidation barriers for the mu-oxo-mu-1,1 peroxodiiron complex are significantly lower than the ones for the mu-oxo-mu-1,2 peroxodiiron complex but still are too high to be considered for catalytic properties. Thus, theory has ruled out two possible peroxodiiron catalysts as oxidants in RNR enzymes and biomimetics and the quest to find the actual oxidant in the enzyme mechanism continues. PMID- 18386301 TI - Analytical optimization of nanocomposite surface-enhanced Raman spectroscopy/scattering detection in microfluidic separation devices. AB - Adding vibrational spectroscopies to the arsenal of detection modes for microfluidics (mufluidics) offers benefits afforded by structurally descriptive identification of separated electrophoretic bands. We have previously applied surface-enhanced Raman spectroscopy (SERS) detection with nanocomposite metal elastomer substrates as a detection mode in mufluidic channels. To create these mufluidic-SERS devices, silver-PDMS substrate regions are integrated into the architecture of a separation chip fabricated from PDMS or glass. Herein, we investigate analytical figures of merit for integrated mufluidic-SERS devices by implementing improvements in fluidic and SERS substrate fabrication as well as data collection strategies. Improvements are achieved by chemical modification of the PDMS channel, increasing effective detection efficiency by minimizing analyte partitioning into nonsensing walls rendering more analyte available to the metallized cover slide of channels and also by uniquely fabricating deep channels that have larger volume to SERS surface area ratios than conventional channels. A method is developed to exploit the inherent concentration profile of analyte material within an electrophoretic band in order to extend the linear dynamic range of detection on the SERS nanostructured surface. This is accomplished by spatially interrogating the Gaussian concentration profile of said bands. The subtleties of this technique give insight into the analytical utility of SERS detection in general. Finally, SERS substrates uniquely created via electron beam lithography with controllable morphologies are integrated into mufluidic-SERS devices to prove feasibility of such a coupling for future work. A separation of endocrine disrupting chemicals in a hybrid SERS nanocomposite-glass device is the capstone of this work. PMID- 18386302 TI - A novel approach for analysis of oligonucleotide-cisplatin interactions by continuous elution gel electrophoresis coupled to isotope dilution inductively coupled plasma mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry. AB - In this work we present a novel approach for in vitro studies of cisplatin interactions with 8-mer oligonucleotides. The approach is based on the recently developed coupling of continuous elution gel electrophoresis (GE) to an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS) with the aim of monitoring the interaction process between this cytostatic drug and the nucleotides. In contrast to existing methods, the electrophoretic separation conditions used here allow both the determination of the reaction kinetics in more detail as well as the observation of dominant intermediates. Two different nucleotides sequences have been investigated for comparison purposes, one containing two adjacent guanines (5'-TCCGGTCC-3') and one with a combination of thymine and guanine (5'-TCCTGTCC-3'), respectively. In order to gain further structural information, MALDI-TOF MS measurements have been performed after fraction collection. This allows for identification of the intermediates and the final products and confirms the stepwise coordination of cisplatin via monoadduct to bisadduct formation. Furthermore, the ICP-MS results were quantitatively evaluated in order to calculate the kinetics of the entire process. PMID- 18386300 TI - Determining under- and oversampling of individual particle distributions in microfluidic electrophoresis with orthogonal laser-induced fluorescence detection. AB - This report investigates the effects of sample size on the separation and analysis of individual biological particles using microfluidic devices equipped with an orthogonal LIF detector. A detection limit of 17 +/- 1 molecules of fluorophore is obtained using this orthogonal LIF detector under a constant flow of fluorescein, which is a significant improvement over epifluorescence, the most common LIF detection scheme used with microfluidic devices. Mitochondria from rat liver tissue and cultured 143B osteosarcoma cells are used as model biological particles. Quantile-quantile (q-q) plots were used to investigate changes in the distributions. When the number of detected mitochondrial events became too large (>72 for rat liver and >98 for 143B mitochondria), oversampling occurs. Statistical overlap theory is used to suggest that the cause of oversampling is that separation power of the microfluidic device presented is not enough to adequately separate large numbers of individual mitochondrial events. Fortunately, q-q plots make it possible to identify and exclude these distributions from data analysis. Additionally, when the number of detected events became too small (<55 for rat liver and <81 for 143B mitochondria) there were not enough events to obtain a statistically relevant mobility distribution, but these distributions can be combined to obtain a statistically relevant electrophoretic mobility distribution. PMID- 18386303 TI - Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis. AB - We present cationized hydroxyethylcellulose (cat-HEC) synthesized in our laboratory as a novel physically adsorbed coating for CE. This capillary coating is simple and easy to obtain as it only requires flushing the capillary with polymer aqueous solution. A comparative study with and without polymers was performed. The adsorbed cat-HEC coating exhibited minimal interactions with basic proteins, providing efficient basic protein separations with excellent reproducibility. Under broad pHs, the amine groups are the main charged groups bringing about a global positive charge on the capillary wall. As a consequence, the cat-HEC coating produced an anodal EOF performance. A comparative study on the use of hydroxyethylcellulose (HEC) and cat-HEC as physically adsorbed coatings for CE are also presented. The separation efficiency and analysis reproducibility proved that the cat-HEC polymer was efficient in suppressing the adsorption of basic proteins onto the silica capillary wall. The long-term stability of the cat-HEC coating in consecutive protein separation runs has demonstrated the suitability of the coating for high-throughput electrophoretic protein separations. PMID- 18386304 TI - Synthesis of copper dendrite nanostructures by a sonoelectrochemical method. AB - Copper dendrites have been prepared by a sonoelectrochemical process from an aqueous solution of Cu(2+) in the presence of polyvinyl alcohol. A SEM image of the morpholohy of the copper formed on the electrode after one electric pulse is presented. A subsequent sonic pulse removes the copper from the electrode surface, cleaning it for the next step. The formation of the dendrites is accounted for by the "drying-mediated self-assembly of nanoparticles" theory. PMID- 18386305 TI - Community consultation for emergency exception to informed consent: how much is enough? PMID- 18386306 TI - Actual and potential drug interactions associated with methadone. AB - OBJECTIVE: To identify and characterize methadone-related drug interactions, as well as factors accounting for the variability in manifesting these interactions clinically. DESIGN: Systematic review of the primary literature. METHODS: Over 200 articles, reports of clinical trials, and case reports were reviewed. Studies and case reports were included if they revealed either quantitative or qualitative methods to identify, evaluate severity of, or compare methadone related drug interactions. RESULTS OF DATA SYNTHESIS: The evidence base associated with methadone drug interactions is underdeveloped in general, as the majority of references found were case reports or case series. Most of the studies and reports focused on inpatients receiving methadone maintenance treatment (MMT) that were between 20 and 60 years of age, taking 200 mg/day of methadone or less. Evidence supporting the involvement of lesser known cytochrome P450 enzymes such as 2B6 is emerging, which may partially explain the inconsistencies previously found in studies looking specifically at 3A4 in vitro and in vivo. Genetic variability may play a role in the pharmacokinetics and pharmacodynamics of many medications, including methadone. CONCLUSIONS: Drug interactions associated with methadone and their clinical significance are still poorly understood in general. Many tertiary drug information references and review articles report interactions associated with methadone in a general sense, much of which is theoretical and not verified by case reports, much less well designed clinical trials. The majority of drug interaction reports that do exist were performed in the MMT population, which may differ significantly from chronic pain or cancer pain populations. PMID- 18386307 TI - Supraventricular tachycardia with simultaneous atrial and ventricular activation: what is the mechanism? PMID- 18386310 TI - Gene symbol: ED1. Disease: Ectodermal dysplasia. PMID- 18386309 TI - New variants of Nav1.5/SCN5A encode Na+ channels in the brain. AB - Exon6A of Nav1.5/SCN5A was first found in the cloning of Nav1.5 from the human neuroblastoma cell line NB-1 (Ou et al., 2005), but its expression in brain and non-brain tissue had not been identified. In this study, we have further investigated this new exon and compared it with exon6 of Nav1.5/SCN5A. Reverse transcription-polymerase chain reaction (RT-PCR) and sequence analysis both confirmed that it is exon6A that encodes Nav1.5 in brain tissue, and it is exon6 that encodes Nav1.5 in non-brain tissue. The expression of exon6A in different parts of the brain is different, with expression levels in the order of hippocampus > cerebral cortex > brain stem > cerebellum. Different expression levels of exon6 in different tissues of Wistar rats were also found. These results suggest that exon6A is unique in encoding the Nav1.5 channels in the central nervous system. In addition, novel alternative splicing of Nav1.5/SCN5A, lacking exon24, was first found in our study. This alternative splicing was also found in other tissues, such as heart, lung and testis. However, the ratio of the two variants changed differently in different types of tissues in developing rats. These results suggest that Nav1.5/SCN5A has a newly identified alternative splicing, and the Nav1.5 channels in the brain are encoded by new variants of Nav1.5/SCN5A. PMID- 18386311 TI - Evidence for habitat fragmentation altering within-population processes in wombats. AB - Habitat fragmentation and associated reduced dispersal of wildlife can lead to an accumulation of related individuals in fragments. The altered kin interactions and amplified chance of inbred matings has profound implications for mating and social systems, and ultimately population persistence. Nonetheless, within fragment population processes are rarely studied. With this aim, we examined relatedness structure in two candidate isolated populations (Kulpara and Scrubby Peak) of southern hairy-nosed wombats (Lasiorhinus latifrons). Wombats were sampled by remote hair-trapping for genotyping at 14 microsatellite loci, enabling individual identification and estimation of space-use and associative behaviour with respect to relatedness. Genetic data indicated that Scrubby Peak was not strongly isolated, against predictions from landscape structure and history. In isolated Kulpara, inhibited female dispersal (normally the dispersing sex) was associated with high population density and altered kin relationships. First, female relatives preferentially coexisted, a radical departure from the previously reported active avoidance of female relatives in continuous habitat. This is consistent with females in altered habitat interacting with more- rather than less-related females to minimize the cost:benefit ratio of proximity to other wombats. Second, inbreeding avoidance appeared to be stronger at Kulpara than in conspecific populations with natural population structures. Although these adaptive behaviours may have contributed to persistence of the Kulpara population in the short term, they are unlikely to ensure its long-term viability in the face of ongoing isolation because they can act only to slow the rate of inbreeding and mitigate some of its negative impacts. PMID- 18386312 TI - Gene symbol: ED1. Disease: Ectodermal dysplasia. PMID- 18386314 TI - Gene symbol: ED1. Disease: Ectodermal dysplasia. PMID- 18386315 TI - Gene symbol: ED1. Disease: Ectodermal dysplasia. PMID- 18386316 TI - Gene symbol: ED1. Disease: Ectodermal dysplasia. PMID- 18386318 TI - Gene symbol: ED1. Disease: Ectodermal dysplasia. PMID- 18386319 TI - Gene symbol: ED1. Disease: X-linked anhidrotic ectodermal dysplasia. PMID- 18386320 TI - Gene symbol: ED1. Disease: X-linked anhidrotic ectodermal dysplasia. PMID- 18386321 TI - Gene symbol: CLCN1. Disease: Myotonia congenita. PMID- 18386322 TI - Gene symbol: EFNB1. Disease: Craniofrontonasal syndrome. PMID- 18386323 TI - Novel human pathological mutations. Gene symbol: GJB1. Disease: Charcot-Marie Tooth disease 1. PMID- 18386324 TI - Novel human pathological mutations. Gene symbol: GJB1. Disease: Charcot-Marie Tooth disease 1. PMID- 18386325 TI - Novel human pathological mutations. Gene symbol: GJB1. Disease: Charcot-Marie Tooth disease 1. PMID- 18386326 TI - Novel human pathological mutations. Gene symbol: MPZ. Disease: Charcot-Marie Tooth disease 1. PMID- 18386327 TI - Gene symbol: EFNB1. Disease: Craniofrontonasal syndrome. PMID- 18386328 TI - Novel human pathological mutations. Gene symbol: MPZ. Disease: Dejerine-Sottas syndrome. PMID- 18386329 TI - Novel human pathological mutations. Gene symbol: MPZ. Disease: Charcot-Marie Tooth disease 1. PMID- 18386330 TI - Novel human pathological mutations. Gene symbol: NOTCH3. Disease: CADASIL. PMID- 18386331 TI - Novel human pathological mutations. Gene symbol: NOTCH3. Disease: CADASIL, exon 2 mutation. PMID- 18386332 TI - Gene symbol: AR. Disease: Androgen insensitivity syndrome. PMID- 18386333 TI - Gene symbol: AR. Disease: Androgen insensitivity syndrome. PMID- 18386334 TI - Novel human pathological mutations. Gene Symbol: CYP21A2. Disease: Non-classic 21 hydroxylase deficiency. PMID- 18386335 TI - Gene symbol: FBN1. Disease: Marfan syndrome. PMID- 18386336 TI - Gene symbol: RHD. Disease: Rhesus negative blood group. PMID- 18386338 TI - Gene symbol: GLA. Disease: Fabry disease. PMID- 18386339 TI - Gene symbol: KRAS. Disease: Cardio-facio-cutaneous syndrome. PMID- 18386340 TI - Gene symbol: BRAF. Disease: Cardio-facio-cutaneous syndrome. PMID- 18386342 TI - Gene symbol: BRAF. Disease: Cardio-facio-cutaneous syndrome. PMID- 18386343 TI - Gene symbol: PPOX. Disease: Porphyria, variegate. PMID- 18386344 TI - Gene symbol: PEO1. Disease: Progressive external ophthalmoplegia. PMID- 18386345 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386348 TI - Human gene mutations. Gene symbol: SLC3A1. Disease: cystinuria. PMID- 18386347 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386349 TI - Gene symbol: LMX1B. Disease: Nail-Patella syndrome. PMID- 18386350 TI - Gene symbol: BEST1. Disease: Best macular dystrophy. PMID- 18386351 TI - Human gene mutations. Gene symbol: SLC3A1. Disease: cystinuria. PMID- 18386352 TI - Human gene mutations. Gene symbol: SLC3A1. Disease: cystinuria. PMID- 18386356 TI - Gene symbol: BEST1. Disease: Best macular dystrophy. PMID- 18386357 TI - Human gene mutations. Gene symbol: MEN1. Disease: MEN1, multiple endocrine neoplasia, Type 1. PMID- 18386358 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386359 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386360 TI - Gene symbol: BEST1. Disease: Best macular dystrophy. PMID- 18386361 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386362 TI - Human gene mutations. Gene symbol: EXT2. Disease: exostoses (multiple) 2. PMID- 18386363 TI - Human gene mutations. Gene symbol: ABCA4. Disease: Stargardt disease. PMID- 18386365 TI - Gene symbol: ABCA4. Disease: Macular dystrophy. PMID- 18386366 TI - Human gene mutations. Gene symbol: GLA. Disease: Fabry disease. PMID- 18386367 TI - Human gene mutations. Gene symbol: EXT1. Disease: exostoses (multiple) 1. PMID- 18386368 TI - Human gene mutations. Gene symbol: CRB1. Disease: late onset retinitis pigmentosa. PMID- 18386369 TI - Gene symbol: ABCA4. Disease: Macular dystrophy. PMID- 18386370 TI - Human gene mutations. Gene symbol: RPS19. Disease: Diamond-Blackfan anaemia. PMID- 18386371 TI - Human gene mutations. Gene symbol: RPS19. Disease: Diamond-Blackfan anaemia. PMID- 18386373 TI - Gene symbol: BEST1. Disease: Best macular dystrophy. PMID- 18386374 TI - Gene symbol: BMPR2. Disease: Pulmonary hypertension, primary. PMID- 18386375 TI - Gene symbol: TSC2. Disease: Tuberous sclerosis. PMID- 18386376 TI - Gene symbol: WFS1. Disease: Wolfram syndrome. PMID- 18386377 TI - Gene symbol: LAMP2. Disease: Glycogen storage disease 2b. PMID- 18386378 TI - Gene symbol: PYGM. Disease: McArdle disease. PMID- 18386380 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386381 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386382 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386383 TI - Gene symbol: PARK2. Disease: Parkinsonism, juvenile, autosomal recessive. PMID- 18386384 TI - Human gene mutations. Gene symbol: SLC3A1. Disease: cystinuria. PMID- 18386385 TI - Human gene mutations. Gene symbol: SLC7A9. Disease: cystinuria. PMID- 18386386 TI - Human gene mutations. Gene symbol: SLC3A1. Disease: cystinuria. PMID- 18386387 TI - Images in neuroscience: psychosis and vertical supranuclear opthalmoplegia. PMID- 18386388 TI - Images in neuroscience: suppression-burst EEG pattern in a neonate with seizures. PMID- 18386390 TI - Microcirculatory alterations of hepatic and mesenteric microcirculation in endotoxin tolerance. AB - LPSs getting access to the circulation of mammalian organisms cause typical systemic inflammatory reactions with symptoms characteristic for acute sepsis. One possibility to attenuate LPS effects is to expose a host to a challenge with low LPS doses, which results in the establishment of "endotoxin tolerance" (ET). Because the microcirculation is of particular importance in LPS action, it seemed of interest to analyze leukocyte-endothelial interactions in the mesentery and liver once endotoxin tolerance has been established and are challenged with LPS. The mesenteric and hepatic microcirculation was investigated by intravital microscopy. After induction of ET LPS, shock was induced by i.v. injection of LPS, and microcirculation of the mesentery and liver was examined. Endotoxin tolerance resulted in reduced ex vivo TNF-alpha synthesis of whole blood. In vivo LPS caused no increase of body temperature. In sinusoids, LPS challenge increased adherence of leukocytes in naive rats, which was almost completely prevented by ET induction. In contrast, in postsinusoidal venules, leukocyte adherence was more intense after ET induction and subsequent to LPS application. Similarly, in postcapillary mesenteric venules, increased adherence of leukocytes after LPS challenge in the ET group was observed. After LPS injection, the endothelial barrier was more disturbed in the nontolerant group when compared with the ET group. Soluble L-selectin and intercellular adhesion molecule were elevated in both ET and untreated rats. Endotoxin tolerance influences leukocyte-endothelial interaction differentially depending on organ and vessel area. PMID- 18386389 TI - Platelet activation by Histophilus somni and its lipooligosaccharide induces endothelial cell proinflammatory responses and platelet internalization. AB - Histophilus somni is a gram-negative coccobacillus that causes respiratory and reproductive disease in cattle. The hallmark of systemic H. somni infection is diffuse vascular inflammation that can lead to an acute central nervous system disease known as thrombotic meningoencephalitis. Previously, we demonstrated that H. somni and its lipooligosaccharide (LOS) activate bovine platelets, leading to expression of P selectin, CD40L, and FasL. Because activated platelets have been reported to induce endothelial cell cytokine production and adhesion molecule expression, we sought to determine if bovine platelets induce proinflammatory and procoagulative changes in bovine pulmonary artery endothelial cells. Endothelial cells were incubated with platelets activated with adenosine diphosphate, H. somni, or H. somni LOS. Incubation with activated bovine platelets significantly increased expression of in adhesion molecules (intercellular adhesion molecule 1, E selectin) and tissue factor, as measured by flow cytometry, real-time polymerase chain reaction, and Western blot analysis. Activated platelets also up regulated expression of endothelial cell IL-1beta, monocyte chemoattractant protein 1, and macrophage inflammatory protein 1alpha as determined by real-time polymerase chain reaction and an IL-1beta enzyme-linked immunosorbent assay. An interesting and surprising finding was that bovine platelets activated by H. somni or its LOS were internalized by bovine endothelial cells as visualized by transmission electron microscopy. This internalization seemed to correlate with endothelial cell activation and morphological changes indicative of cell stress. These findings suggest that activated platelets might play a role in promoting vascular inflammation during H. somni infection. PMID- 18386391 TI - The preoptic anterior hypothalamic area mediates initiation of the hypotensive response induced by LPS in male rats. AB - The mechanism responsible for the initiation of endotoxic hypotension is not fully understood, although it is often attributed to a direct effect of LPS and other vasoactive mediators on the vasculature. Alternatively, recent evidence raises the possibility that endotoxic hypotension may be initiated through a central mechanism. Previous studies have shown that LPS initiates fever, sickness behavior, and other aspects of the inflammatory response through a neural pathway that sends peripheral inflammatory signals to the preoptic anterior hypothalamic area (POA). It is also well known that the POA plays a role in the regulation of cardiovascular function, but its involvement in LPS-induced hypotension has not been examined previously. Therefore, the aim of the present paper was to investigate whether the initial abrupt fall in arterial pressure evoked by LPS in septic shock is mediated by the POA. LPS (1 mg/kg, i.v.) administration to halothane-anesthetized or conscious rats lowered arterial blood pressure by 24.8+/-2.9 and 25.1+/-5.8 mmHg, respectively. Bilateral lidocaine (2%; 1 microL) injection into the POA, but not the lateral hypothalamus, prevented the hypotension evoked by LPS entirely in both anesthetized and conscious animals. Remarkably, this blockade significantly inhibited the second, delayed fall in arterial pressure induced by LPS, and simultaneously decreased TNF-alpha plasma levels. Together, these data indicate that the initial phase of endotoxic hypotension is mediated by the POA and suggest that the initiation of the hypotensive response induced by LPS can be essential for the development of the late fall in blood pressure. PMID- 18386392 TI - Cetuximab: head and neck cancers: for second-line treatment, in combination with radiotherapy. PMID- 18386393 TI - Epinephrine versus dopamine to treat shock in hypoxic newborn pigs resuscitated with 100% oxygen. AB - Shock and tissue hypoperfusion are common after asphyxia. We compared systemic and regional hemodynamic effects of epinephrine and dopamine in the treatment of shock and hypotension in asphyxiated newborn piglets resuscitated with 100% oxygen. Twenty-four piglets (1-3 days old; weight, 1.4-2.6 kg) were acutely instrumented to measure cardiac index (CI), carotid, mesenteric and renal arterial blood flows, and mean systemic (SAPs) and pulmonary arterial pressures (PAPs). Piglets had normocapnic alveolar hypoxia (F(IO2)=0.08-0.10) for 50 min and reoxygenated with F(IO2)=1.0 for 1 h then F(IO2)=0.21 for 3.5 h. After 2 h reoxygenation, either dopamine (2 microg kg(-1) min(-1)) or epinephrine (0.2 microg kg(-1) min(-1)) was given for 30 min in a blinded randomized manner, which was then increased to maintain SAP (within 10% of baseline, pressure-driven dose) for 2 h. Hypoxia caused hypotension (SAP, 44%+/-3% of baseline), cardiogenic shock (CI, 41%+/-4%), and metabolic acidosis (mean pH, 7.04-7.09). Upon reoxygenation, hemodynamic parameters immediately recovered but gradually deteriorated during 2 h with SAP at 45+/-1 mmHg, CI at 74+/-9% of baseline, and pH 7.32+/-0.03. Low doses of either drug had no significant systemic and renal hemodynamic response. Epinephrine (0.3-1.5 microg kg(-1) min(-1)) for 2 h increased SAP and CI (with higher stroke volume) and decreased pulmonary vascular resistance (with reduced PAP-SAP ratio), whereas the responses with dopamine (10 25 microg kg(-1) min(-1)) were modest. Low-dose epinephrine improved mesenteric and carotid arterial flows, whereas the pressure-driven doses of epinephrine and dopamine increased carotid and mesenteric arterial flows, respectively. To treat shock in asphyxiated newborn piglets resuscitated with 100% oxygen, epinephrine exhibits an inotropic action compared with dopamine, whereas both catecholamines can increase carotid and mesenteric perfusion. PMID- 18386394 TI - Inhaled and nasal steroids: mental disorders. AB - Depression, mania, euphoria and delirium are known adverse effects of oral and injectable steroids. But they can also occur during steroid inhalation or nasal administration. PMID- 18386395 TI - Nimesulide and hepatitis: death, transplantation; regulatory failures. PMID- 18386396 TI - Omalizumab: anaphylactic shock: sometimes occurs more than one hour after the injection. AB - Sometimes occurs more than one hour after the injection. PMID- 18386397 TI - Efalizumab: inflammatory neuropathies: higher than expected. PMID- 18386398 TI - Glitazones: fractures: avoid using glitazones. PMID- 18386399 TI - Varenicline: myocardial infarction: nicotine replacement is a better option. PMID- 18386400 TI - Editing the ERJ: an observational study. PMID- 18386401 TI - [Tako-tsubo-resembling syndrome with atypical clinical presentation]. PMID- 18386402 TI - NY: Five-year-old asthmatic kindergartner died: no basis for suit against school nurse or school. PMID- 18386403 TI - [Deworming of dogs and cats]. PMID- 18386404 TI - [BAFTA Study-comparison of aspirin and warfarin in preventive maintenance of stroke in the oldest persons with atrial fibrillation]. PMID- 18386405 TI - Pediatric bipolar disorder is a controversial diagnosis. PMID- 18386406 TI - Growing evidence to support early intervention in early onset bipolar disorder? PMID- 18386408 TI - Vaccine study pipeline faces ethical and other challenges. PMID- 18386407 TI - [When and why in treatment for cardiogenic shock is an anesthetist wanted?]. PMID- 18386409 TI - Study finds excellent clinical ART responses from Kenyans. Toxicities were top reason for switch. PMID- 18386410 TI - Evolution of reproductive systems in the genus Silene. AB - The genus Silene contains both hermaphrodite, gynodioecious and dioecious species, dioecy being represented in three sections of the genus. To locate the events of change of reproductive systems, we compared ITS sequences of 22 species of Silene chosen throughout the whole genus, and four putative outgroup species. Gynodioecy, which is the most common reproductive system within the genus Silene and in closely related genera such as Saponaria and Dianthus, is proposed to be ancestral in the genus. Dioecy has evolved at least twice: once in the section containing S. latifolia, and once in the clade containing S. otites and S. acaulis ssp. bryoides. Evolution towards hermaphroditism, associated with evolution of selfing has also occurred at least twice, in S. gallica and S. comica. PMID- 18386411 TI - A past: a revolution in public ethics. PMID- 18386412 TI - The spread of primary schooling in Sub-Saharan Africa: implications for fertility change. PMID- 18386413 TI - Women's diaries as life-savings: who decides whose life is saved? The journals of Eugenie de Guerin and Elisabeth Leseur. PMID- 18386414 TI - [Lev Nikolaevich Gumilev (1912-92): biologism and Eurasianism in Russian thought]. PMID- 18386416 TI - [From experimental embryology to a genetics of development: from Hans Spemann to Antonio Garcia-Bellido]. PMID- 18386415 TI - Transplanting and rooting workers in London and Brussels: a comparative history. PMID- 18386417 TI - Safe sects? Dynamic religion and AIDS in South Africa. PMID- 18386418 TI - Abstracts of the Joint Annual Meeting of the ADSA, PSA, AMPA, ASAS. July 8-12, 2007, San Antonio, Texas, USA. PMID- 18386419 TI - "The doctor from the university is at the door": methodological reflections on research with non-Aboriginal adoptive and foster mothers of Aboriginal children. PMID- 18386421 TI - A feeding frenzy, or feeling friendsy? Events after the death of Diana, Princess of Wales. PMID- 18386420 TI - William Percy Rogers (1914-1997). PMID- 18386422 TI - Marriage and bridewealth in a matrilineal society: the case of the Tonga of Southern Zambia: 1900-1996. PMID- 18386423 TI - Displacement and disease: epidemics and ideas about malaria in Matabeleland, Zimbabwe, 1945-1996. PMID- 18386424 TI - Association between periodontal disease and coronary heart disease. AB - There were increasing studies on the association of periodontal diseases with coronary heart disease (CHD) in the recent 20 years. This article reviewed the evidence supporting the association between periodontal disease and CHD, the possible mechanisms explaining the association, and the possible effect of periodontal treatment on the risk of CHD. In general, it is suggested that periodontal disease especially, periodontitis is modestly associated with CHD. Besides there are some common risk factors, such as smoking, stress, elder age, male gender and low socioeconomic status, between these two diseases, long chronic periodontal infections by periodontal pathogens, the systemic acute-phase response and host immuno-inflammatory response to the exposures of periodontal infection appear to be the important mechanisms for connecting the periodontal disease and CHD. Periodontal treatment can decrease the infection of periodontal pathogens and therefore, reduce the systemic inflammatory burden. It was observed in some studies that periodontal treatment could reduce the serum inflammatory biomarkers such as C-reactive protein and improve endothelial function. Even though the studies on effect of periodontal interfere on CHD are still limited now, it appears hopeful that periodontal treatment could reduce the risk of CHD and therefore become one of the preventing strategies for CHD. PMID- 18386425 TI - [Neurophysiopathology of eating disorders]. PMID- 18386426 TI - [Eating disorders and maternity-infertility]. PMID- 18386427 TI - [The role of fathers in eating disorders]. PMID- 18386428 TI - Mambila demography from archival sources. PMID- 18386429 TI - Relative cohort size: source of a unifying theory of global fertility transition. PMID- 18386430 TI - Food for growth: trends in Indonesia's food supply, 1880-1995. PMID- 18386431 TI - An unusual cause of a difficult extubation. PMID- 18386432 TI - Effect of ovomocoide on human basophil activation. PMID- 18386433 TI - "In vivo" and "in vitro": tests in the diagnosis of trichophyton allergy. AB - A group of 100 patients with Trichophyton allergy and a control of group of 100 without fungal allergy have been compared in order to evaluate the diagnostic value of skin prick and intradermal tests and assay of Trichophyton specific IgE. The evaluation of sensitivity, specificity efficacy, positive and negative predictive values suggests that skin tests in two steps, prick and intradermal and research of specific IgE must be used to a better diagnostic approach. In cases were allergy is not cleared by antifungal therapy specific immunotherapy is worthwhile to be tried with a great percentage of success. PMID- 18386434 TI - FEF75 in asthma management. AB - RATIONALE: Diagnosis of asthma is based on the presence of symptoms (clinical diagnosis) supported by lung function measurements, such as forced expiratory volume in one second (FEV1) and peak expiratory flow (PEF) variability. Recently, it has been reported that the forced expiratory flow at 25-75 of the pulmonary volume might be considered as a possible marker of early bronchial impairment in patients with allergic rhinitis. OBJECTIVES: The aim of this retrospective study was to identify the most reliable spirometric parameter for the follow up of intermittent allergic asthma. METHODS: Data from 108 outpatients with intermittent allergic asthma was studied. The spirometric parameters before and after 1 week of short aerosol therapy were analyzed. Patients were divided into three groups according to basal FEV1 values: those with all spirometric values normal, those with reduced FEV1 and those with normal FEV1 but reduced FEF75. RESULTS: There was a good correlation between the spirometric parameters analysed. FEF75 showed a greater significant reduction than FEV1 and PEF. Moreover, 26 of 38 patients with normal FEV1 (68 %) showed a reduction in FEF75. No significant differences were observed between FEV1 and PEF. After short aerosol therapy, FEV1 improved in 93% of patients, but many (61%) still had a reduced FEF75 value, thus the correlation between the two parameters decreased (from r= 0.68 to r = 0.40). CONCLUSIONS: On the basis of these observations, FEF75 should be reconsidered a usefull spirometric parameter for allergic asthma follow up. PMID- 18386435 TI - Role of breast feeding in primary prevention of asthma and allergic diseases in a traditional society. AB - BACKGROUND: The fact that breastfeeding may protect against allergic diseases remains controversial, with hardly any reports from developing countries. Prolonged breastfeeding was shown to reduce the risk of allergic and respiratory diseases. AIM: The aim of this study was to assess the relationship between breastfeeding and the development of childhood asthma and allergic diseases in Qatari children at age 0-5 years. Additionally, this study investigated the effect of prolonged breastfeeding on the allergic diseases in a developing country. DESIGN: This is a cross sectional survey. SETTING: Well baby clinics and Pediatric clinics in the 11 Primary Health Care Centers and Hamad General Hospital, Hamad Medical Corporation, State of Qatar. SUBJECTS: A multistage sampling design was used and a representative sample of 1500 Qatari infants and pre-school children with age range of 0-5 years and mothers aged between 18 to 47 years were surveyed during the period from October 2006 to September 2007 in Qatar. Out of the 1500 mothers of children, 1278 mothers agreed to participate in this study with the response rate of 85.2%. METHODS: A confidential, anonymous questionnaire was completed by the selected subjects assessing breastfeeding and allergic diseases. Questionnaires were administered to women who were attending Primary Health Centers for child immunization. Questionnaire included allergic rhinitis, wheezing, eczema, and additional questions included mode and duration of breastfeeding, tobacco smoke exposure, number of siblings, family income, level of maternal education, parental history of allergies. Univariate and multivariate statistical methods were performed for statistical analysis. RESULTS: More than half of the infants (59.3%) were exclusively breastfed, followed by infants with partial breastfeeding (28.3%) and artificial fed (12.4%). There was a significant difference found across these three categories of infants in terms of their age groups, smoking status of father, socio-economic status and parental consanguinity. Asthma (15.6%), wheezing (12.7%), allergic rhinitis (22.6%), and eczema (19.4%) were less frequent in exclusive breast fed children, compared to infants with partial breast feeding and formula milk. Ear infection (P = 0.0001) and eczema (P = 0.007) were found significant in infants with the history of maternal atopy, while asthma (P = 0.0001) and allergic rhinitis (P = 0.015) were found significant in infants with the history of paternal atopy. The main factors associated with mode of feeding were mothers having first baby, asthmatic mother and parental history of allergic rhinitis. The risk of allergic diseases, eczema, wheeze and ear infection in particular, were lower in children with prolonged breast feeding (>6 months) than in those with short-term breast feeding duration (<6 months). CONCLUSION: The current study indicates that exclusive breast-feeding prevents development of allergic diseases in children. The main factors associated with breastfeeding for allergic diseases were being the first baby, maternal history of asthma, and parental history of allergic rhinitis. The study findings opens a big avenue for interventional role of breastfeeding. Therefore, we recommend breastfeeding is as one possible way to reduce the risk of onset asthma and allergic diseases in developing countries. PMID- 18386436 TI - A case of severe allergy to pine-nut. AB - A case is reported of a patient who showed an immediate allergic reaction after ingestion of pine nut at the age of 38. Subsequently she developed acute allergic symptoms also by inhalation of the allergen, without food ingestion. Finally, the patient showed an acute systemic allergic reaction immediately after skin testing with fresh pine nut. PMID- 18386438 TI - Should nurses be fingerprinted and subjected to background checks? PMID- 18386439 TI - Self-employed nurses and patient care. PMID- 18386440 TI - Traveling south--or west--for the winter. PMID- 18386441 TI - VRE. How you can stop the spread of this drug-resistant organism. PMID- 18386442 TI - Finding their niche. Why men choose nursing. PMID- 18386443 TI - Understanding the world of children with autism. PMID- 18386444 TI - Fitness makeover 2008: the real world. PMID- 18386445 TI - Reflections on caring. Fighting words. PMID- 18386448 TI - Our struggle to attain professional recognition. PMID- 18386449 TI - Understanding teams and teamwork. PMID- 18386450 TI - Industrial nursing. 1929. PMID- 18386451 TI - Facilitating interprofessional education and practice. AB - Collaborative patient-centred care has the potential to address serious issues in the Canadian health-care system such as those related to increasing complexity of care; patient safety and access; and recruitment and retention of health human resources. This approach involves teams of health professionals working together to provide more coordinated and comprehensive care to clients. It places priority on the preferences of the patient and fosters respect for the skills and perspectives of all health-care providers. Interprofessional education at the undergraduate, graduate and practice levels is essential for facilitating the transition to team-based care. The author presents the rationale for collaborative care and describes an interprofessional education project at Memorial University of Newfoundland that is preparing students and health professionals for this groundbreaking change in practice. PMID- 18386453 TI - Celeste Johnston: at the forefront of pain research. Interview by Patrick McCloskey. PMID- 18386452 TI - A closer look at the "supervision" and "direction" of certified registered nurse anesthetists. AB - A growing shortage of anesthesiologists in Canada has prompted discussion of how anesthesia provision can be expanded. Canadian anesthesiologists generally support a team approach in which physicians supervise alternative providers. In the U.S., nurses have worked as anesthetists for over 150 years, and their experiences of different models of anesthesia provision provide valuable insights into the potential pitfalls of the team approach as well as the benefits of autonomous nurse anesthetist roles. The authors conducted a qualitative study of the anesthesia team and the role of nurse anesthesia practice in the U.S., and here they present a summary of some of their preliminary findings and the implications for evolving models of care in Canada. Inefficiencies associated with the medical supervision and direction of Certified Registered Nurse Anesthetists are discussed. PMID- 18386454 TI - A strategy for building interprofessional teams. PMID- 18386455 TI - Is interprofessional practice rhetoric or reality? PMID- 18386456 TI - Cervical screening in Hungary: why does the "English model" work but the "Hungarian model" does not? AB - A comparison has been made between the English practice and the "Hungarian model" of cervical screening. In England, until 1986, extensive opportunistic screening was the practice, but--as it had no effect on cervical cancer mortality- afterwards, the screening policy was changed to be strictly in line with international recommendations. On the other hand, in Hungary, the "old practice" has been petrified: gynaecologists are the "gatekeepers", a "gynaecological examination completed with smear-taking for cytology" makes up the screening strategy. Although in the frame of a National Public Health Programme all the prerequisites for nationwide organised screening have been provided, and an up-to date screening strategy declared, 20-times as many smears are taken and analysed outside as inside the programme, and the efforts have had no impact on cervical cancer mortality. This is because "old habits die hard". There is an urgent need to reconsider the screening strategy, and to reorganise the cervical screening practice in Hungary. PMID- 18386457 TI - Potential therapeutic vaccine strategies and relevance of the immune system in uterine cervical cancer. AB - The interaction of HPV with the immune system has been studied, but the results are still inconclusive for several reasons. Until now, we have not been able to understand the mechanisms of immune system regulation in the uterine cervix. HPV infection does not unleash an inflammatory response, and consequently an efficient and specific immune response against the virus. Moreover, an understanding of HPV infection and local immune response is indispensable for the development of new bioactive drugs and therapies for patients with both non invasive and invasive tumors, mainly for patients that do not present regression with radiotherapy or chemotherapy or in whom the tumors are surgically unresectable. The aim of this review is to provide support in understanding potential mechanisms used by the immune system to destroy neoplastic cells, comparing the immunotherapy used in cancer and discussing the possibility of developing new drugs based on these mechanisms of action. PMID- 18386458 TI - Immunohistochemical bcl-2 expression, p53 overexpression, PR and ER status in endometrial carcinoma and survival outcomes. AB - Immunohistochemical expression of bcl-2, p53, PR and ER in cases with endometrial carcinomas arrayed on a tissue microarray (TMA) was tested and correlated with clinicopathologic features, overall survival (OS), cancer-related survival (CRS) and disease-free survival (DFS). Seventy-seven patients with endometrial cancer were reviewed. Slides were evaluated by two pathologists blinded to patient clinical characteristics and survival data. Mean age of patients was 62.5 years (range 35-80), median follow up 60 months (range 9-120). Seventy-nine percent of patients were FIGO Stage I; 39% of the cases showed bcl-2 cytoplasmic staining and its expression was significantly correlated with low-grade tumor differentiation and age < or = 60 years. Nuclear p53 overexpression was detected in 23.4% of the cases and was significantly correlated with advanced stages (IIB IV), non-endometrioid histology, nodal metastasis and advanced age (> 60 years). PR and ER were positive in 63.6% and 30% of the cases, respectively. Analysis of p53 overexpression and bcl-2 expression in relationship with PR and ER status showed a direct correlation between bcl-2 expression and PR positivity (p = 0.001). In a multivariate analysis FIGO staging was the only clinicopathologic parameter independently correlated with DFS. In conclusion p53 overexpression was directly associated with unfavorable clinicopathologic factors such as advanced stage, histologic subtype, advanced patient age and nodal metastasis. Bcl-2 expression was related with younger age, favorable grade and PR expression by tumor cells. Patient survival was not related to the tested biomarkers. PMID- 18386459 TI - L1 (CAM) (CD171) in ovarian serous neoplasms. AB - PURPOSE OF THE INVESTIGATION: The evaluation of L1 (CAM) as a tumor progression marker and as a prognostic factor in serous ovarian tumors. METHODS: L1 (CAM) protein expression was assessed by immunohistochemistry and Western blot in serous ovarian tumors [cystadenomas (n = 20), borderline tumors (n = 14) and carcinomas (n = 47)], and was correlated with stage,grade, progression-free survival time (PFS) and overall survival. RESULTS: L1 (CAM) immunoreactivity correlated significantly with stage and grade. It increased from benign tumors to early carcinomas and to advanced stage carcinomas progressively and significantly. In Stage III G3 carcinoma patients, low L1 (CAM) expressing tumors exhibited better response to chemotherapy and were associated with statistically significantly longer PFS (p = 0.002). CONCLUSION: L1 (CAM) expression represents a novel diagnostic marker in serous ovarian neoplasms that shows characteristics of tumor progression. L1 expression was associated with chemotherapy response. PMID- 18386460 TI - Recurrent cervical carcinoma after radical hysterectomy and pelvic lymph node dissection: a study of 32 cases. AB - PURPOSE OF INVESTIGATION: To investigate the characteristics of patients with recurrent cervical carcinoma after radical hysterectomy and pelvic lymph node dissection (RHND), and to evaluate the effect of clinical and surgical pathologic factors on the outcome of these patients. METHODS: Data from the files of 32 patients with recurrent cervical carcinoma after RHND managed at the Soroka Medical Center from 1962 through 2005 were analyzed. RESULTS: These 32 patients represent a recurrence rate of 25.4%. The median recurrence-free interval was 19.3 (range, 1-106) months. The prevailing signs and symptoms were obstructive nephropathy, sacral pain and bowel obstruction. Sixteen (50%) patients had loco regional recurrence alone, 12 (27.5%) loco-regional plus distant recurrence, and four (12.5%) distant recurrence alone. Treatment modalities included radiotherapy, chemotherapy and various surgical procedures. The 5-year survival rate was 35%, with 22 (68.7%) of the patients dead of disease at the end of follow-up. Univariate analysis demonstrated a significant worsening in survival with each of the following factors: loco-regional plus distant recurrence (p = 0.010), positive pelvic lymph nodes (p = 0.010), tumor size > or = 3 cm (p = 0.013), positive lymph vascular space involvement (p = 0.017) and RHND without bilateral salpingo-oophorectomy (p = 0.042). In a multivariate analysis, extent of recurrent disease (locoregional alone versus loco-regional plus distant recurrence) and pelvic lymph node status (negative vs positive) at RHND were the only significant predictors of survival. Uremia was the most common cause of death. CONCLUSIONS: Recurrent cervical carcinoma after RHND is a grave disease with unfavorable prognosis. In both univariate and multivariate analyses, extent of recurrent disease and pelvic lymph node status at RHND were significant predictors of survival. PMID- 18386461 TI - Concentrations of follicle stimulating hormone are increased in ovarian tumor fluid: implications for the management of ovarian cancer. AB - PURPOSE OF INVESTIGATION: Significant progress has been made in recent years in the understanding of the mechanisms postulated by the gonadotropin theory of ovarian carcinogenesis. In the present study we compare FSH concentrations between serum and fluid from cysts or the rectouterine pouch of patients with epithelial tumors and non-neoplastic lesions. METHODS: We enrolled 277 patients. They were divided into five groups: I (n = 44)--ovarian cancer patients, II (n = 16)--borderline tumors, III (n = 40)--benign epithelial cystadenomas, IV (n = 137)--non-neoplastic lesions and V (n = 22)--admitted for "second-look" laparoscopy. RESULTS: There were any significant differences between FSH concentrations in serum and tumor fluid in patients with ovarian cancer (36.46 vs 28.11 mIU/ml) and borderline epithelial tumors (31.5 vs 22.7 mIU/ml). For benign cystadenomas the respective concentrations were 28.96 mIU/ml in serum and 6.93 mIU/ml in tumor fluid in these groups p < 0.0000001. The same highly significant differences were found in non-neoplastic lesions (24.97 vs 4.77 mIU/ml), p < 0.0000001. Patients who underwent "second-look" laparoscopy demonstrated significant differences (p < 0.05) as FSH concentration in serum and peritoneal fluid when neoplastic cells were not disclosed, but the difference was not significant (p = 0.752) when fluid from the rectouterine pouch was positive for carcinoma cells. CONCLUSIONS: The results of our study can reflect an ineffective tumor: blood barrier and easy diffusion of gonadotropins into the tumor tissue. Local reduction of FSH levels through administration of GnRH analogs may in some clinical situations produce clear therapeutic benefits for the management of ovarian malignancies. PMID- 18386462 TI - Prognostic factors for the development of vaginal intraepithelial neoplasia. AB - OBJECTIVE: To identify risk factors for the appearance of vaginal intraepithelial neoplasia (VAIN). MATERIAL AND METHODS: A total of 485 women with abnormal cytologies were followed over three years (2003-2006). They underwent cytolology and colposcopy, and testing for human papillomavirus virus (HPV) infection. If the colposcopy was atypical, a biopsy was performed. RESULTS: A total of 256 women were treated: 161 by cone biopsy, 103 by LLETZ, 12 by repeat conization, and 44 by total hysterectomy. In eight cases VAIN was diagnosed following hysterectomy. The average age at which VAIN appeared was 49.8 years (age range 39 61). Hysterectomy was indicated in two cases of cervical cancer, four cases of persistent high-grade cervical SIL, and two cases of recurrent high-grade cervical SIL. The mean time for the appearance of VAIN following hysterectomy was 3.8 years (range 1-9 years). Of these eight women, seven had HPV infections at high risk for carcinogenesis. CONCLUSIONS: Long-term follow-up cytology is necessary for women treated for high-grade SIL, even after hysterectomy, because of the increased risk of a primary vaginal VAIN lesion, especially in women with high-risk HPV infection. PMID- 18386463 TI - Results of postoperative radiotherapy in the treatment of uterine sarcomas: a retrospective analysis of 46 patients. AB - PURPOSE: The aim of this study was to evaluate treatment outcome, survival data and prognostic factors in patients with uterine sarcoma treated by postoperative radiotherapy. MATERIALS AND METHODS: The records of 46 patients treated between 1993 and 2003 were reviewed. Median age was 55 (range 31-75). There were 21 mixed mullerian tumors, 12 leiomyosarcomas, 11 endometrial stromal sarcomas and two adenosarcomas. According to FIGO classification 65.2% were Stage I, 17.4% Stage II, 13% Stage III and 4.3% Stage IV. All patients received external radiotherapy with 1.8 Gy daily fractions up to 50.4-64 Gy (median 50.4 Gy). Intracavitary brachytherapy was applied to 39 patients. Twelve patients received adjuvant chemotherapy. RESULTS: Median follow-up time was 48 months (6-144 months). Seventeen patients (37%) developed distant metastases and one patient had local failure. Five-year overall, disease-free and local recurrence-free survival rates were 57.8%, 60.5% and 97.8%, respectively. Univariate analysis demonstrated that stage (p = 0.011), histologic subtype (p = 0.010), tumor size (p = 0.044), positive peritoneal cytology (p = 0.006) and the use of chemotherapy (p = 0.005) had a significant effect on overall survival. Prognostic factors influencing disease-free survival were stage (p = 0.009), positive peritoneal cytology (p = 0.000) and the use of chemotherapy (p = 0.002). The only prognostic factor affecting local control was stage (p = 0.000). CONCLUSION: Postoperative radiotherapy seems to be an effective adjuvant treatment providing high local control rates in uterine sarcomas. However its efficacy should be clarified by randomized trials. The important prognostic factors influencing the treatment results were stage, histologic subtype, tumor size and positive peritoneal cytology. PMID- 18386464 TI - Migrant women and cervical cancer: background of a prevention study. AB - The study was scheduled in order to organize a program of prevention against cervical cancer in female migrants in Rome, and therefore to facilitate access to appropriate preventive oncological facilities for discriminated women. Moreover, the study will also investigate the risk factors and social conditions (HPV subtypes, sexual behavior, smoking habits) of such women since their migration to Italy. This is scientific and cultural background of a longitudinal, observational study on the cervical cancer risk in Roman migrant population. By means of a mother language questionnaire (with the presence of a cultural mediator) it will be possible to achieve data on social conditions and the new life-style. An HPV-testing (HC2) combined with Pap-test (with further genotype distribution) will be performed in all women enrolled in the study. Further diagnostic/therapeutic decisions will depend on the results of both tests. Scientific results are expected in the next two years, but an increasing of cancer prevention awareness among female migrant populations is expected from the beginning of the program. The present study was aimed at culturally appropriate intervention strategies to limit the disparities that migrants usually suffer in most of the developed Western nations in respect to the native counterparts. PMID- 18386465 TI - Outcome of uterine clear cell carcinomas compared to endometrioid carcinomas and poorly-differentiated endometrioid carcinomas. AB - OBJECTIVES: Our aim was to compare the survival between patients with clear cell carcinoma (CC) and patients with endometrioid carcinoma (EC). METHODS: Through the population-based Geneva Cancer Registry, we identified 1,380 resident women diagnosed with uterine cancer between 1970 and 2000. We excluded those with papillary serous endometrial carcinoma and uterine sarcomas. We categorized patients as CC (n = 32, 2.8%) or EC (n = 1,145, 97.2%). Uterine cancer-specific survival rates were calculated by Kaplan-Meier analysis. We used Cox proportional hazards analysis to compare uterine cancer mortality risks between groups, and adjusted these risks for other prognostic factors. RESULTS: CC patients presented with a more advanced stage at diagnosis than EC patients (p = 0.002). Compared to women with EC, women with CC had a significantly greater risk of dying from their disease (hazard ratio [HR] 2.9, 95% confidence interval (95% CI) 1.7-4.9). After adjustment for age, stage and adjuvant chemotherapy, the risk of dying from uterine cancer was still significantly higher for CC patients (HR 2.0, 95% CI 1.2 3.4). By univariate analysis, the risk of dying of endometrial cancer was not significantly higher in CC patients than in patients with poorly-differentiated EC (HR 1.3, 95% CI 0.7-2.3). CONCLUSION: This population-based investigation shows that patients with CC have a poorer outcome than those with EC. Studies to determine the role of adjuvant treatment in CC patients are needed. PMID- 18386466 TI - Sociodemographic and clinicopathologic characterization of cervical cancers in northern Nigeria. AB - OBJECTIVE: To evaluate the sociodemographic and clinicopathological characteristics of patients with cervical cancer seen in a tertiary referral center in northern Nigeria. MATERIALS AND METHODS: Between January 2002 and December 2004, 70 consecutive patients with histologically confirmed cervical cancer, with a median age of 48 years (range, 30-75 years), were interviewed on the basis of a structured pro forma. RESULTS: Of these patents, 39 (56%) had had no formal education, and 36 (51%) were unemployed housewives. Sixty (86%) had become sexually active before 17 years of age; 44 (63%) were in polygamous families, and 25 (36%) patients were in at least a second marriage. There was an average of 6.8 live births per patient. Vaginal bleeding was seen in all patients, and 55 (79%) had vaginal discharges; 50 (71%) had a bulky cervical mass, and 46 (66%) presented with at least Stage IIIA disease. Squamous cell carcinoma was the commonest histology. The three HIV-seropositive patients were young and had advanced disease. CONCLUSION: Sociodemographic factors, such as low socioeconomic level, early age at first sexual intercourse and multiple sexual partners, place women at high risk of developing cervical cancer in northern Nigeria. Late presentation with advanced disease predominates. PMID- 18386467 TI - FIGO Stage I endometrial carcinoma: evaluation of lung metastases and follow-up. AB - PURPOSE: The aim of our study was to evaluate the incidence of lung metastases in the follow-up of women submitted to surgery for endometrial carcinoma, in particular for FIGO Stage I which is the lowest risk stage for this metastatic site. METHODS: The study was conducted on 210 patients affected by FIGO Stage I endometrial cancer in the years 1990 to 2005 distributed as follows: 35 patients with Stage IA (limited to the endometrium), 150 patients with Stage IB (invasion up to and including half the myometrial thickness), 25 patients with Stage IC (invasion greater than half the myometrial thickness). They underwent follow-up. RESULTS: Only one patient out of the group studied has developed lung metastasis six years after surgery. She was staged as FIGO IB (T1b Mx G1). CONCLUSION: We are still following the cases and evaluating the biological behavior of this specific endometrial carcinoma and its reaction to further therapies. We are also looking for possible clinical characteristics in disagreement with those reported in the literature, which would thus make it necessary to reconsider the prognosis and therapy of this stage of disease. PMID- 18386468 TI - TV sonographic assessment in postmenopausal women with bleeding. AB - The aim of this study was to evaluate retrospectively the usefulness of transvaginal sonography for the detection of endometrial disease in postmenopausal women with bleeding. This study involved 275 postmenopausal women aged 47-81 years (median 62). None of them were on hormone replacement therapy and all had had amenorrhea for more than one year. Concerning the age of the study patients, we confirm that endometrial cancer occurs at any age, but more commonly in ages above 58 years. Transvaginal sonography was performed in all women. About 89.2% of malignant diseases were discovered in the study women whose endometrial thickness was above 4 mm, but we also found endometrial cancer in 10.2% of the cases in women whose endometrial thickness was below 4 mm. In postmenopausal symptomatic women premalignant or malignant causes of bleeding can not be excluded with just transvaginal ultrasound. PMID- 18386469 TI - Risk factors for cervical cancer in China: a case-control study. AB - OBJECTIVES: To determine the prevalence of human papillomavirus types and investigate the risk factors for cervical cancer in Hubei, China. METHODS: We conducted a case-control study to investigate risk factors. RESULTS: HPV DNA was detected in 94.55% of patients with cervical carcinoma, and 23.64% of control subjects. The most common HPV type in cervical cancer was HPV type 16 (81.82%), followed by HPV 58 (6.36%). HPV infected patients have a higher risk of developing cervical carcinoma, which is 75.79 times more than non-infected people. The other risks were age at first intercourse (p = 0.017) and number of live births (p = 0.032). A history of previous cytologic screening was associated with a substantial reduction in risk (p = 0.001). CONCLUSIONS: The three principal reasons that Hubei has a high rate of women developing cervical carcinoma are HPV infection, age at first sexual intercourse and number of live births. Cervical cytology screening provides efficacious protection. PMID- 18386470 TI - Knowledge and interest of Turkish women about cervical cancer and HPV vaccine. AB - OBJECTIVE: We aimed to evaluate the knowledge and interest level of Turkish women about HPV, HPV vaccines and cervical cancer using a questionaire. METHOD: A 25 item questionnaire was distributed to women in three different cities located in separate sociocultural locations. RESULTS: At the closure of the study 143 women responded and returned the survey. Of the participants 62.2% (89) had a university degree, 36.4% (52) a high school education, and 1.4% (2) had lower school degrees; 98.5% of the women would consent to have their daughter vaccinated for HPV and 94.7% would consent to have their son vaccinated if vaccine provided prevention against cancer and related diseases. However in both cases women gave importance to the "cost" - unless vaccine could be free. On logistic regression analyses none of the variables (i.e., questions) in the survey predicted women's willingness to accept the vaccine for themselves or their children. CONCLUSIONS: Women in Turkey would be willing to have themselves and their children receive HPV vaccine against cervical cancer and related diseases. PMID- 18386471 TI - Relationship between risk factors and tumor stage in breast cancer patients in a university hospital--Brazil. AB - PURPOSE: To verify the relationship between clinical variables and tumor stage in breast cancer. METHODS: This retrospective study (1998 to 2001) analyzed data of 176 women with breast cancer attending a university hospital. Patients were divided into groups according to the clinicopathological variables studied. RESULTS: The disease had a similar frequency at age under 50 years (44.3%) or above (55.7%) 50 years. Stage II was more frequent. Most patients were white (69.9%), non-smokers (69.3%) and were not using oral contraceptives (71%). Stages 0-II were mainly detected in the white (74.8%) vs non-white (60.4%) group. Monthly breast self-exams were performed by 62.5% of women, in which earlier stages (0, I) were more frequently detected than in those who did not perform self-exams (27.3% vs 12.1%, p = 0.01). CONCLUSION: Breast cancer occurred mainly in white women in Stage II, and with similar frequency at age under or over 50 years. Breast self-exam was associated with early detection of the disease. PMID- 18386472 TI - Cold-knife conization versus the loop electrosurgical excision procedure for treatment of cervical dysplasia. AB - INTRODUCTION: Classical conization is a standard procedure for treatment of cervical lesions. Conization with loop diathermy is well established and lesions can be excised in more than 90% of cases. OBJECTIVE: To compare two methods of conization for the treatment of cervical dysplasia. METHOD: The study included 172 patients who had conization for diagnosed cervical dysplasia. A retrospective analysis was conducted on incidence of complications and presence of dysplasia on the specimen edges after classical conization compared to conization with loop diathermy. The possibilities for analyzing specimen edges were reviewed. RESULTS: A significantly higher incidence of complications was found among patients who underwent classical conization compared to those who had the loop diathermy procedure. The loop procedure is sufficient for treatment of cervical dysplasias. CONCLUSION: The authors suggest loop diathermy conization as the method of choice for treatment of cervical dysplasia. PMID- 18386473 TI - Recurrence of granulosa cell tumor 25 years after initial diagnosis. Report of a case and review of the literature. AB - Granulosa cell tumors (GCTs) are rare functional sex cord-stromal ovarian tumors constituting approximately 2-3% of all ovarian malignancies. They are characterized by low malignant potential, local spread, late recurrence and high survival rates. We report a case of recurrent ovarian GCT in a 60-year-old woman 25 years after the initial diagnosis. The patient underwent surgical resection of the pelvic masses and refused to receive any adjuvant treatment, considering the late recurrence and high survival rates of this tumor. This case illustrates an example of a very late recurrence and emphasizes the importance of the extended follow-up required for these patients. PMID- 18386474 TI - Ovarian cancer after kidney transplantation. AB - Malignancies are one of the major causes of morbidity and mortality in transplant patients. The incidence is progressively increasing either because of the increased age transplant patients and the increase of immunosuppressive therapy or the increased follow-up range post-transplantation. The main causes of increased tumor incidence in transplant patients with respect to the general population are the reduced immunosurveillance and the high incidence of infections due to oncogenic viruses. This problem might become more and more serious in the near future due to the introduction of new immunosuppressive strategies that significantly extend allograft survival. A case of ovarian cancer in a kidney transplant patient is described. Attention is focused on the potential dual role of immunosuppressive therapy in the development of malignancies in transplant patients. PMID- 18386475 TI - Solitary ovarian mass: a case of metastatic malignant melanoma. AB - Malignant melanoma involving the ovaries is rare and in most cases metastatic in origin. We present a case of ovarian malignant melanoma presenting as a large adnexal mass in a patient with no previous history of malignant melanoma. PMID- 18386476 TI - A case of synchronous relapse of breast cancer and uterine mullerian adenosarcoma post tamoxifen in a premenopausal woman. AB - PURPOSE & METHODS: We report a case of a 42-year-old multigravida, premenopausal woman with breast carcinoma, who presented after four years of use of adjuvant tamoxifen with synchronous liver, bone, and lung metastasis of breast cancer with mullerian adenosarcoma. RESULTS: Immunohistochemical stains on the uterine tumor for estrogen and progesterone receptors showed positivity for both epithelial and stromal cells, actin, and desmin while the proliferative index (MIB-1) showed positivity for stromal cells only. The patient underwent a hysterectomy followed by palliative chemotherapy. She died 14 months after her relapse because of progressive disease (cerebral, bone, liver and lung metastases). CONCLUSION: Our case is the only one reported in the literature with synchronous relapse of breast adenocarcinoma and a Mullerian adenosarcoma. Moreover, it is one of the rare cases occurring in a premenopausal woman since all except two cases were postmenopausal. PMID- 18386477 TI - Leiomyosarcoma of the uterine corpus with ovarian metastases in a 28-year-old woman: case report. AB - A case of invasive leiomyosarcoma of the uterine corpus with right ovarian metastases in a 28-year-old woman is presented. The patient was submitted to surgery including total abdominal hysterectomy with left salpingo-oophorectomy, dissection of the pelvic and paraaortal lymph nodes and fixation of the right ovary to the psoas muscle. Postoperative radiation therapy was applied. A year after treatment, the patient was well. PMID- 18386478 TI - Endometrial metastasis of a primitive neuroendocrine ovarian carcinoma: management and treatment of a case. AB - BACKGROUND: Neuroendocrine tumours are a heterogeneous group of separate clinico pathological entities which have a common characteristic, i.e., expression of potential endocrine differentiation. In the ovary, the term "neuroendocrine" relates mainly to widely known carcinoids, but it may also be applied to rare neuroendocrine carcinomas as non-small cell type and small cell carcinomas of pulmonary type. In the literature only 11 cases of primary ovarian non-small cell neurendocrine carcinomas have been described and ten of these were associated with a surface epithelial ovarian tumour. Small cell neuroendocrine carcinoma of the ovary is a rare malignant tumour of the ovary. Advanced small cell carcinoma of the ovary is a very aggressive tumour with an overall poor prognosis and unfavourable outcome. CASE REPORT: The case reported is unique in the literature because the authors describe a rare case of endometrial metastasis of a primary ovarian non-small cell neurendocrine carcinoma without any surface epithelial ovarian tumour association. The tumour invaded up to less than half of the myometrium. The first symptoms were related to endometrial metastasis as metrorrhagia and pelvic pain while the asymptomatic presence of primary ovarian carcinoma was not acknowledged with physical examination, routine biochemistry, tumour markers, blood count and traditional transvaginal greyscale ultrasound. CONCLUSION: Magnetic resonance and three-dimensional (3D) ultrasonography with power Doppler are a great help in the diagnosis of ovarian localisation but only immunohistochemistry on histological material can provide a correct diagnosis. Immunohistochemistry expression of Ki67 is a useful marker of malignancy. Due to the rarity of this neoplasm, a general consensus for optimal treatment has yet to emerge. The reported biological aggressiveness of these tumours prompts combined treatment with radical surgery and adjuvant polychemotherapy. PMID- 18386479 TI - Isoflavonoid glycosides and rotenoids from Pongamia pinnata leaves. AB - Chromatographic separation of a 70% aqueous methanol extract (AME) of Pongamia pinnata (Linn.) Pierre (Leguminosae) leaves has led to the isolation of two new isoflavonoid diglycosides, 4'-O-methyl-genistein 7-O-beta-D-rutinoside (2) and 2',5'-dimethoxy-genistein 7-O-beta-D-apiofuranosyl-(1"'-->6")-O-beta-D glucopyranoside (6), and a new rotenoid, 12a-hydroxy-alpha-toxicarol (5), together with nine known metabolites, vecinin-2 (1), kaempferol 3-O-beta-D rutinoside (3), rutin (4), vitexin (7), isoquercitrin (8), kaempferol 3-O-beta-D glucopyranoside (9), 11,12a-dihydroxy-munduserone (10), kaempferol (11), and quercetin (12). Their structures were elucidated on the basis of chemical and spectroscopic analyses. PMID- 18386480 TI - Ornicorrugatin, a new siderophore from Pseudomonas fluorescens AF76. AB - From a pyoverdin-negative mutant of Pseudomonas fluorescens AF76 a new lipopeptidic siderophore (ornicorrugatin) could be isolated. It is structurally related to the siderophore of Pseudomonas corrugata differing in the replacement of one Dab unit by Orn. PMID- 18386481 TI - Stereochemistry of lignans in Phaleria macrocarpa (Scheff.) Boerl. AB - Phaleria macrocarpa (Scheff.) Boerl., a member of the Thymelaeaceae, is traditionally used in Indonesia as medicinal plant against cancer. In this context, we isolated the lignans pinoresinol, lariciresinol and matairesinol from different parts of this plant. The enantiomeric composition of these lignans was determined by chiral column analysis. Pinoresinol and lariciresinol were mixtures of both enantiomers with (79 +/- 4)% and (55 +/- 6)% enantiomeric excess for the (-)-enantiomers, respectively, whereas matairesinol was found as pure (+) enantiomer. PMID- 18386483 TI - Antifungal activity of coumarins. AB - The antifungal activity of 40 coumarins was tested against the fungal strains: Candida albicans (ATCC 14053), Aspergillus fumigatus (ATCC 16913) and Fusarium solani (ATCC 36031), using the broth microdilution method. Osthenol showed the most effective antifungal activity among all the compounds tested, with a MIC value of 125 microg/ml for Fusarium solani and 250 micro/ml for Candida albicans and Aspergillus fumigatus. The antifungal potential of this prenylated coumarin can be related to the presence of an alkyl group at C-8 position. PMID- 18386482 TI - Extraction of condensed tannins from Mexican plant sources. AB - Contents of total polyphenols, condensed tannins and proanthocyanidins, and their stability to various pH values and temperatures were studied in Mexican blueberry, cuautecomate fruit, garambullo fruit, aubergine, coffee pulp and residues of black grapes. Several aqueous extracts, obtained through a one-pass extraction process, were analyzed using liquid chromatography in order to quantify the condensed tannin (proanthocyanidin) content responsible for their antioxidant activity and colour. All tested samples included high proanthocyanidin contents demonstrating that these Mexican fruits and vegetables are good sources of natural antioxidants, and they all could be considered as excellent functional foods due to their bioactivity measured as the condensed tannin level. PMID- 18386484 TI - Activities of 2,4-dihydroxy-6-n-pentylbenzoic acid derivatives. AB - Esters of 2-hydroxy-4-methoxy-6-n-pentylbenzoic acid (2-8) (methyl, ethyl, butyl, pentyl, isopropyl, sec-butyl and benzyl), olivetol (9), methyl, ethyl, butyl perlatolates (10-12), 2,4-dihydroxy-6-n-pentylbenzoic acid (15), and methyl and ethyl esters of (15) were prepared through structural modifications of perlatolic acid (1) with the aim to detect new antifungal and antibacterial substances and also to evaluate the toxicity by the brine shrimp lethality assay against Artemia salina. The antifungal assays were carried out against the fungus Cladosporium sphaerospermum through the bioautography method, and methyl 2,4-dihydroxy-6-n pentylbenzoate (13) showed the highest antifungal activity (2.5 yg). Olivetol (9) and 2,4-dihydroxy-6-n-pentylbenzoic acid (15) are also potent inhibitors of the growth of the fungus (5.0 microg). Except for methyl (10), the ethyl (11) and butyl (12) perlatolates were less active than perlatolic acid (1). The activities presented by methyl (2) and ethyl (3) 2-hydroxy-4-methoxy-6-n-pentylbenzoates and methyl (13) and ethyl (14) 2,4-dihydroxy-6-n-pentylbenzo-ates suggest that compounds with a free hydroxy group in the aromatic ring (C-4) have a more pronounced effect against C. sphaerospermum. Antibacterial activities were tested by the disc diffusion method using pathogenic strains of S. aureus and E. coli. The compounds were weakly active with inhibition zones between 9-15 mm. The 2 hydroxy-4-methoxy-6-n-pentylbenzoic esters 2-8 and alkyl perlatolates 10-12 were selective against E. coli. Perlatolic acid (1) and methyl 2-hydroxy-4-methoxy-6-n pentylbenzoate (2) were the most active with LD50 values of 24.1 microM and 27.2 microM, respectively. The other compounds were not toxic to Artemia salina larvae. PMID- 18386485 TI - Phenolic constituents from the wood of Morus australis with cytotoxic activity. AB - A new methylated flavonol, 5,7,2',4'-tetrahydroxy-3-methoxyflavone (1), had been isolated from the methanol extract of the wood of Morus australis, along with nine known compounds, kuwanon C (2), morusin (3), morachalcone A (4), oxyresveratrol (5), 4'-(2-methyl-2-buten-4-yl)oxyresveratrol (6), moracins M (7) and C (8), alboctalol (9), and macrourin B (10). The structures of these compounds were determined based on spectral evidence, including UV, IR, NMR, and mass spectra. Cytotoxic properties of compounds 1-10 were evaluated against murine leukemia P-388 cells. The prenylated stilbene 6 and 2-arylbenzofuran 8, and morusin (3) were found to have strong cytotoxic effects with IC50 values of 6.9, 8.7, and 10.1 microM, respectively. PMID- 18386486 TI - Antimicrobial activity of the extract and isolated compounds from Baccharis dracunculifolia D. C. (Asteraceae). AB - Baccharis dracunculifolia D.C. (Asteraceae) is the most important plant source of the Brazilian green propolis. Since propolis is known for its antimicrobial activity, the aim of this work was to evaluate the antimicrobial activities of B. dracunculifolia and some of its isolated compounds. The results showed that the leaves extract of B. dracunculifolia (BdE) presents antifungal and antibacterial activities, especially against Candida krusei and Cryptococcus neoformans, for which the BdE showed IC50 values of 65 microg mL(-1) and 40 microg mL(-1), respectively. In comparison to the BdE, it was observed that the green propolis extract (GPE) showed better antimicrobial activity, displaying an IC50 value of 9 microg mL(-1) against C. krusei. Also, a phytochemical study of the BdE was carried out, affording the isolation of ursolic acid (1), 2a-hydroxy-ursolic acid (2), isosakuranetin (3), aromadendrin-4'-methylether (4), baccharin (5), viscidone (6), hautriwaic acid lactone (7), and the clerodane diterpene 8. This is the first time that the presence of compounds 1, 2, and 8 in B. dracunculifolia has been reported. Among the isolated compounds, 1 and 2 showed antibacterial activity against methicillin-resistant Staphylococcus aureus, displaying IC50 values of 5 microg mL(-1) and 3 microg mL(-1), respectively. 3 was active against C. neoformans, showing an IC50 value of 15 microg mL(-1) and a MIC value of 40 microg mL(-1), while compounds 4-8 were inactive against all tested microorganisms. The results showed that the BdE, similar to the GPE, displays antimicrobial activity, which may be related to the effect of several compounds present in the crude extract. PMID- 18386487 TI - Antimicrobial activity of 2,5-dihydroxy-3-methyl-1,4-benzoquinone from Embelia schimperi. AB - Chromatographic separation of an ethyl acetate extract from Embelia schimperi led to the isolation of a new compound identified as 2,5-dihydroxy-3-methyl-1,4 benzoquinone (1) on the basis of spectroscopic and physical data. The plant's crude extract and pure compound 1 were assayed for in vitro antimicrobial activity against clinical strains of Salmonella spp., Proteus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Cryptococcus neoformans, Shigella dysentriae and Staphylococcus aureus. Disc diffusion method was used and zones of inhibition, after respective incubation periods, were used to quantify antimicrobial activity. Standard antibiotics namely: augmentin, cotrimoxazole, gentamycin, tetracycline and lyncomycin were used as controls. The crude extract was inactive while the pure compound 1 showed significant activities against Salmonella spp., Proteus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Cryptococcus neoformans, Shigella dysentriae and Staphylococcus aureus with zones of inhibition ranging from 10-20 mm. The most sensitive microorganism was P aeruginosa while C. neoformans was insensitive to both the crude extract and compound 1. PMID- 18386488 TI - Termiticidal activity of diterpenes from the roots of Euphorbia kansui. AB - Five ingenane compounds, 1-5, kansuinins A and B, isolated from Euphorbia kansui, and their derivatives 7 and 9 were tested for termiticidal activity against the Japanese termite, Reticulitermes speratus. At 72 hours after treatment, the ingenane compounds 1 to 5 caused 100% mortality in R. speratus at 50, 25 and 12.5 microg/disk, respectively, except for compound 1, which gave a mortality rate of (93.06 +/- 5.56)% at 12.5 microg/disk. At 36, 48 and 60 hours after treatment, compounds 1 to 5 showed more termiticidal activity than kansuinins A and B and their derivatives. The kansuinins showed no or only slight activity against termites in the filter paper bioassay under the conditions tested compared with a solvent control. PMID- 18386489 TI - Antinematodal activities of ingenane diterpenes from Euphorbia kansui and their derivatives against the pine wood nematode (Bursaphelenchus xylophilus). AB - Under the bioassay-guided method, two diterpenes, 3-O-(2",3"-dimethylbutanoyl)-13 O-dodecanoylingenol (1) and 3-O-(2",3"-dimethylbutanoyl)-13-O-decanoylingenol (2) isolated from Euphorbia kansui, showed a pronounced antinematodal activity against the nematode Bursaphelenchus xylophilus at the same minimum effective dose (MED) of 5 microg per cotton ball and still displayed antinematodal activity at a dose of 2.5 microg per cotton ball. Compounds 3-6 were obtained, and the structure of the new compound 6 was elucidated based on 1D- and 2D-NMR analyses and physicochemical data. Preliminary structure-biological activity relationships of ingenane-type compounds were deduced. PMID- 18386490 TI - Free radical scavengers, antioxidants and aldose reductase inhibitors from Camptosorus sibiricus Rupr. AB - Flavonoids and organic acids were recommended in the literature as the main active constituents of Camptosorus sibiricus Rupr. Assay-guided fractionation led to the isolation of 9 flavonoids and 8 phenolic acids. All compounds were tested for DPPH scavenging activity, SOD-like and aldose reductase inhibition. Among them, compounds 1, 2, 3, 5, 6, 7, 8, 9, 11, 15 showed activities. The most active free radical scavenger and antioxidant was compound 8, while compound 1 exhibited strong inhibiting activity of aldose reductase. The structure-activity relation was dicussed briefly. PMID- 18386491 TI - Flavonoids in Helichrysum pamphylicum inhibit mammalian type I DNA topoisomerase. AB - DNA topoisomerases are important targets for cancer chemotherapy. We investigated the effects of a methanolic extract of Helichrysum pamphylicum on mammalian DNA topoisomerase I via in vitro plasmid supercoil relaxation assays. The extracts manifested a considerable inhibition of the enzyme's activity in a dose-dependent manner. We also performed a HPLC analysis to identify the flavonoid content of the H. pamphylicum extract and tested the identified flavonoids; luteolin, luteolin-4-glucoside, naringenin, helichrysinA and isoquercitrin, on DNA topoisomerase I activity. The measurement of the total antioxidant capacity of the flavonoid standards suggested that the topoisomerase inhibition might be correlated with the antioxidant capacity of the plant. PMID- 18386492 TI - Anti-herpesvirus activities of Pseudomonas sp. S-17 rhamnolipid and its complex with alginate. AB - The rhamnolipid biosurfactant PS-17 and its complex with the polysaccharide alginate, both produced by the Pseudomonas sp. S-17 strain, were studied for their antiviral activity against herpes simplex virus (HSV) types 1 and 2. They significantly inhibited the herpesvirus cytopathic effect (CPE) in the Madin Darby bovine kidney (MDBK) cell line. The investigations were carried out according to the CPE inhibition assay protocol. The suppressive effect of the compounds on HSV replication was dose-dependent and occurred at concentrations lower than the critical micelle concentration of the surfactant. The 50% inhibitory concentration (IC50) of rhamnolipid PS-17 was 14.5 microg/ml against HSV-1 and 13 microg/ml against HSV-2. The IC50 values of the complex were 435 microg/ml for HSV-1 and 482 microg/ml for HSV-2. The inhibitory effects of the substances were confirmed by measuring the infectious virus yields with the multicycle virus growth experimental design as well: deltalog CCID50 of 1.84-2.0 against the two types of herpes simplex viruses by rhamnolipid PS-17 (20 microg/ml), and a strong reduction of the HSV-2 virus yield under the effect of the alginate complex at a concentration of 450 microg/ml. The results indicate that rhamnolipid PS-17 and its alginate complex may be considered as promising substances for the development of anti-herpetic compounds. PMID- 18386493 TI - Biotransformation of indole derivatives by mycelial cultures. AB - Biotransformation of tryptophan to tryptamine and 3-methyl-indole by Psilocybe coprophila was performed. On the other hand, Aspergillus niger was able to transform tryptophan to 5-hydroxy-tryptophan. P. coprophila biotransformed 5 hydroxy-tryptophan to 5-hydroxytryptamine. These results prove once more that fungi are good tools to establish hydroxyindole derivatives. PMID- 18386494 TI - Outdoor and indoor cultivation of Spirulina platensis in the extreme south of Brazil. AB - Water supplemented with 10% or 20% (v/v) of Zarrouk medium was used to cultivate Spirulina platensis in closed and open bioreactors under controlled conditions (30 degrees C, 32.5 micromol m(-2) s(-1), 12 h light/dark photoperiod) and in a greenhouse (9.4 to 46 degrees C, up to 2800 micromol m(-2) s(-1), variable day length photoperiod) using different initial biomass concentrations (X0) in the extreme south of Brazil (32.05 degrees S, 52.11 degrees W). Under controlled conditions the maximum specific growth rate (micromax) was 0.102 d(-1), the biomass doubling time (t(d)) was 6.8 d, the maximum dry biomass concentration (Xmax) was 1.94 g L(-1) and the maximum productivity (Pmax) was 0.059 g L(-)1 d( 1), while the corresponding values in the greenhouse experiments were micromax = 0.322 d(-1), t(d) = 2.2 d, Xmax = 1.73 g L(-1) and Pmax = 0.112 g L(-1) d(-1). Under controlled conditions the highest values for these parameters occurred when X0 = 0.15 g L(-1), while in the greenhouse X0 = 0.4 g L(-1) produced the highest values. These results show that the cultivation of S. platensis in greenhouses in the extreme south of Brazil is technically viable and that the S. platensis inoculum and the concentration of Zarrouk medium can be combined in such a way as to obtain growth and productivity parameters comparable, or superior, to those occurring in bioreactors under controlled conditions of temperature, illuminance and photoperiod. PMID- 18386495 TI - Ginsenoside contents in Panax quinquefolium organs from field cultivation. AB - Quantitative composition of saponins (ginsenosides) in Panax quinquefolium originating from field cultivation in Poland and harvested at the beginning of the growing season, during the blossoming period and at the end of growth was determined. A colourimetric method gave lower values compared to results of HPLC analysis, but the tendency of change in the content of saponins was similar in both instances. Ginsenoside amounts in P. quinquefolium organs changed depending on the specific time during the vegetation season the samples were taken. This study found that the highest content of these metabolites--66 mg/g d. w.-- occurred in the roots of the plant at the time of bloom. Two among the six metabolites examined in our study were dominant independently of the vegetation season. These were Rb1 and Re, with values of 25.4-33.8 mg/g d. w. and 16.4-19.7 mg/g d. w., respectively. PMID- 18386496 TI - Artemisinin production by shoot regeneration of Artemisia annua L. using thidiazuron. AB - An efficient in vitro method for multiple shoot bud induction and regeneration has been developed in Artemisia annua L. using leaf and stem explants in various concentrations and combinations of plant growth regulators to evaluate the frequency of regeneration. The sources of explants as well as plant growth regulators in the medium were found to influence the multiple shoot induction. The result shows that the stem segment cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg/l thidiazuron (TDZ) gave a perfect shoot formation (100%) and good shoot multiplication (57 shoots/explant) after 2 weeks of culture. Healthy regenerated shoots were elongated and rooted in MS medium without hormones. The artemisinin content in plants regenerated from stem explants using 0.1 mg/l TDZ was (3.36 +/- 0.36) microg/mg dry weight and two-fold higher than that of in vitro grown plants of the same age [(1.73 -/+ 0.23) microg/mg DW]. This system exhibited a potential for a rapid propagation of shoots from the stem explant and makes it possible to develop a clonal propagation of A. annua. PMID- 18386497 TI - Changes in foliar proline concentration of osmotically stressed barley. AB - The amino acid proline is accumulated in plant tissues in response to a variety of stresses. The existence of two routes for its biosynthesis is well documented. However, little is known about the contribution of each pathway to the accumulation of free proline under stress conditions. In the present study young barley plants were subjected to osmotic stress by treating their roots with 25% polyethylene glycol. Prior to stress imposition roots were incubated for 24 h in nutrient solution containing proline or one of its metabolic precursors: glutamate and ornithine. Free proline quantity in the leaves was measured before and after stress. Relative water content (RWC) was used as a measure of the plant water status. Foliar proline levels showed a significant increase in ornithine- and proline-pretreated plants compared to the control. Nevertheless, no considerable changes in leaf RWC were observed. It was shown that before stress application only ornithine but not glutamate was immediately metabolized to proline. Under stress conditions, however, both precursors were converted into proline. The possible role of this amino acid in the processes of post stress recovery is discussed. PMID- 18386498 TI - Characterization of an aminopeptidase and a proline iminopeptidase from cabbage leaves. AB - Aminopeptidase, preferring phenylalanine-p-nitroanilide as substrate, and proline iminopeptidase, highly-specific for proline-p-nitroanilide, were isolated from cabbage leaves (Brassica oleraceae var. capitata). As pH optima, 7.2-7.5 for aminopeptidase activity and 8.0-8.5 for proline iminopeptidase were determined. Both peptidases were strongly inhibited by p-chloromercuribenzoic acid, heavy metal ions and urea. The molecular weights were determined by gel filtration to be 56 and 204 kDa, respectively. The iminopeptidase was decomposed during SDS electrophoresis to four subunits of 50 kDa. Minor impurities of myrosinase associated protein (approximately 70 kDa) were found in both preparations. Preliminary data of their amino acid sequences showed similarities to those of aminopeptidases N (family M1) and proline iminopeptidases (family S33). PMID- 18386499 TI - Photobiological control of crop production and plant diseases. AB - Plants, as well as fungi, use ambient sunlight as information to regulate photomorphogenetic processes. The photobiological control of this information showed that the development of photobiological greenhouse plastic covers simulates a photonic information that leads to a physiological enhancement of plant productivity and fungal disease control, thus minimizing the need for the use of agrochemicals. The main characteristics of these photobiological greenhouse plastic covers are the high transmission of photosynthetically active radiation (PAR, 400-700 nm) combined with an increase of the factor zeta = RL(655 665 nm)/FRL(725-735 nm), which affects the cellular phytochromic equilibrium phi = Pfr/(Pfr + Pr) and regulates the photosynthetic activity and therefore the plant productivity. Additionally, increase of the spectral ratios from the transmitted light: BL(420-500 nm)/nearUV(290-370 nm) and BL(420-500 nm)/ FRL(725 735 nm), cause mainly the induction of biochemical, physiological and morphological responses, regulated by cryptochromes in plants (e.g. inflorescence and infructescence) and mycochrome in fungi (e.g. inhibition of sporulation). In the present work, comparative studies with randomly selected greenhouse plastics showed that small changes in the above-mentioned "photobiological" parameters raise the productivity of tomato plants and inhibit the sporulation of several isolates of the fungal pathogen Botrytis cinerea. Thus, a model for the photoregulation of these two phenomena in greenhouses is proposed. PMID- 18386500 TI - An improved method for the isolation of total RNA from Avicennia germinans leaves. AB - Isolation of high-quality RNA of Avicennia germinans L. tissue is difficult due to high levels of phenols and other substances that interfere when using conventional procedures for the isolation. These substances not only decrease the yield but also the quality of RNA is almost poor. We present here a simple RNA protocol and fast methodology that effectively removes these contaminating substances without affecting the yield. The protocol developed is based on the SDS/phenol method with modifications including beta-mercaptoethanol to prevent oxidation of phenolic complexes, and phenol/chloroform extraction is introduced to remove proteins, genomic DNA, and secondary metabolites, and co-precipitated polysaccharides. Both A260/A230 and A260/A280 absorbance ratios of isolated RNA were around 2 and the yield was about 0.3 mg g(-1) fresh weight. Good-quality total RNA from leaves of Avicennia germinans could be easily isolated within 2 h by this protocol which avoided the limitation of plant materials and could provide total RNA for all kinds of further molecular studies. PMID- 18386501 TI - Effect of cell culture on 18S rRNA gene sequences in the cultural course of Taxus chinensis cells. AB - Cell culture is an effective technology for taxol production. This paper discusses the effect of Taxus cell cultures on the 18S rRNA gene sequences based on the phylogenetic analysis of cultured T. chinensis cells and related species. The phylogenetic tree is reconstructed using the maximum parsimony method and the relative rate test to test the hypothesis of a molecular clock. The phylogenetic analysis indicates that cell culture changes the phylogenetic position of cultured T. chinensis cells. More than that, the 18S rRNA gene of cultured T chinensis cells has a faster rate of substitution than that of T. chinensis. With T. media as reference, the divergence time of the cultured T. chinensis cells is 7 Ma (million years) more than that of the T. chinensis cells based on the 18S rRNA gene sequences. PMID- 18386502 TI - Molecular analysis of phenol-degrading microbial strains. AB - In an attempt to estimate the occurrence of phenol hydroxylase-related gene sequences we performed a dot blot hybridization assay with DNA from phenol utilizing Trichosporon cutaneum R57 strain NBIMCC 2414 and microbial isolates from different wastewaters. The used oligonucletides were homologous to the 5' end of TORPHD locus (NCBI)-coding phenol hydroxylase in Trichosporon cutaneum ATCC 46490 and to the 5'-end of TORCCMLE locus (NCBI)-coding cis,cis-muconate lactonizing enzyme in Trichosporon cutaneum ATCC 58094. Two microbial strains, Escherichia coli JM 109 and Lactobacillus acidophilus ATCC 4356, incapable to degrade phenol were used as negative controls. We established the presence of hybridization with both used oligonucleotide probes in T. cutaneum R57 and T. cutaneum ATCC 46490 yeast strains. The experiments implemented with microbial isolates obtained from three industrialized areas in Bulgaria showed that 7 of them may carry sequences hybridizing with a phenol hydroxylase oligonucleotide probe. A subsequent hybridization test for the cis,cis-muconate-lactonizing enzyme showed that only 3 of them displayed a positive signal. Lactobacillus acidophilus ATCC 4356 and Escherichia coli JM 109 strains' DNA used as negative controls in the experiments did not reveal any sequence similarity to the both applied oligonucleotides. The partial nucleotide sequences of 16S rDNAs of the isolated strains C1 and K1 obtained as PCR products were determined and sequenced. A comparison of these nucleotide sequences with similar sequences in NCBI Data Bank indicated that both C1 and K1 strains are closely related to the genera Acinetobacter and Burkholderia. PMID- 18386503 TI - Identification of parasporin genes in Vietnamese isolates of Bacillus thuringiensis. AB - Four genes encoding parasporins, cytotoxins preferentially killing human cancer cells in vitro, were isolated from four Vietnamese strains of Bacillus thuringiensis. Nucleotide sequence analysis revealed that: (1) three genes fall into the two known classes, ps1Aa and ps1Ab, and (2) another one belongs to ps1Ac, a novel gene class established in this study. Upon proteolytic activation, parasporal protein of the organism with ps1Ac exhibited strong cytocidal activity against human cancer cells, HeLa and Hep G2, but not to non-cancer normal cells, UtSMC and HC. PMID- 18386504 TI - Isolation and characterization of a new Arthrospira strain. AB - A filamentous microorganism, morphologically similar to the cyanobacterium Arthrospira, was isolated from Mangueira Lagoon in Brazil, from which Arthrospira has not previously been isolated. Random amplified polymorphic DNA (RAPD) comparison with the standard Arthrospira platensis strains LEB 52 and Paracas indicated that the organism isolated was an Arthrospira isolate, which we denominated strain LEB 18. The RAPD analysis showed conserved sequences which indicated that the three strains belonged to the same genus, and were all Arthrospira species, but there were sufficient differences between them suggesting that they were separate strains. The strain LEB 18 was cultivated in undiluted Zarrouk medium and in 60% and 20% (v/v) Zarrouk medium diluted with sterilized Mangueira Lagoon water (MLW) using illuminance rates of 32.5, 45.5 and 58.5 micromol m(-2) s(-1) according to a complete 32 factorial design with a triplicate central point. The strains LEB 52 and Paracas were cultived in the conditions central point. Our new isolate produced the highest specific growth rate (Umax = 0.22 d(-1)) in 60% Zarrouk medium diluted with MLW and illuminated with 58.5 micromol m(-2) s(-1) and the highest protein content (86.0% w/w). PMID- 18386505 TI - Capillary electrophoresis study on the dimeric SOD enzyme in the presence of ascorbic acid. AB - A Notable decrease of the peak intensity of the capillary electrophoregram due to the dimeric SOD molecule was observed when a solution containing copper(II) chloride and ascorbic acid was added to the SOD solution, indicating that the capillary electrophoresis method is useful to detect the dissociation of the dimeric SOD molecule in solution, and that dissociation of the dimeric SOD molecule is induced by the presence of hydrogen peroxide. The present results may give reasonable countermeasures towards the sporadic amyotrophic lateral sclerosis in future. PMID- 18386506 TI - Facile uptake of manganese(III) by apotransferrin: possible origin of manganism. AB - We have investigated the mechanism of manganese ion uptake by apo-transferrin using a capillary electrophoresis method, and obtained clear evidence that oxidation state +3 and the binuclear unit of a manganese chelate are essential factors for the facile uptake by apotransferrin, similar to that observed for Fe(III) chelates. These results may give valuable information to understand the pathogenesis of manganism and to develop new countermeasures for the neurotoxicity by manganese ions. PMID- 18386507 TI - Bag culture: a method for root-root co-culture. AB - A method named "bag culture" was developed for coculturing of Linum persicum (section Syllinum) and L. austriacum (section Linum) hairy roots. For this propose L. austriacum and L. persicum hairy root cultures were established using Agrobacterium rhizogenes in McCown medium. L. persicum hairy roots in bags (1 mm2 mesh) were successfully grown together with L. austriacum hairy roots. The amounts of podophyllotoxin (PTOX) and 6-methoxypodophyllotoxin (MPTOX) produced by L. persicum hairy root cultures were detected using HPLC. The results indicated that the amounts of both lignans and growth indexes of the two hairy roots decreased, that may be partly due to a competition between the two types of culture in using precursors of biosynthetic metabolites and the amount of culture medium which is available for each hairy root. However, MPTOX (0.17 g/100 g DW) and PTOX (0.02 g/100 g DW) levels of the L. persicum single culture in bag were significantly higher than of the other cultures which may be due to the immobilization effect of the bag. PMID- 18386508 TI - Honouring our debt to history. PMID- 18386509 TI - Recording nursing highlights through the pages of Kai Tiaki. PMID- 18386510 TI - Viewing nursing's present concerns through the lens of history. PMID- 18386511 TI - It's all about making a difference. PMID- 18386512 TI - Negotiating an unstable ladder. PMID- 18386513 TI - Down the decades. PMID- 18386514 TI - Preserving kawa whakaruruhau--what is Kai Tiaki's role? PMID- 18386515 TI - Kai Tiaki--its role in NZNO's industrial profile. PMID- 18386517 TI - Penalising nurses hurts whole families. PMID- 18386516 TI - Nurses, trade unionists and journalists--a troubled trinity? PMID- 18386518 TI - Questioning nursing ethics. PMID- 18386519 TI - Health reform: finding the way. PMID- 18386521 TI - Assessing risk in aged care mental health. PMID- 18386522 TI - A little reassurance goes a long way. PMID- 18386523 TI - ATSDR evaluation of potential for human exposure to zinc. AB - As part of its mandate, the Agency for Toxic Substances and Disease Registry (ATSDR) prepares toxicological profiles on hazardous chemicals found at Comprehensive Environmental Response, Compensation and Liability Act (CERCLA) National Priorities List (NPL) sites that have the greatest public health impact. These profiles comprehensively summarize toxicological and environmental information. This article constitutes the release of portions of the toxicological profile for zinc. The primary purpose of this article is to provide interested individuals with environmental information on zinc that includes production data, environmental fate, potential for human exposure, analytical methods and a listing of regulations and advisories. PMID- 18386524 TI - ATSDR evaluation of potential for human exposure to tungsten. AB - As part of its mandate, the Agency for Toxic Substances and Disease Registry prepares toxicological profiles on hazardous chemicals found at Comprehensive Environmental Response, Compensation and Liability Act, National Priorities List sites that have the greatest public health impact. These profiles comprehensively summarize toxicological and environmental information. This article constitutes the release of portions of the Toxicological Profile for Tungsten. The primary purpose of this article is to provide interested individuals with environmental information on tungsten that includes production data, environmental fate, potential for human exposure, analytical methods and a listing of regulations and advisories. PMID- 18386525 TI - ATSDR evaluation of health effects of tungsten and relevance to public health. AB - The Agency for Toxic Substances and Disease Registry prepares toxicological profiles, as part of its mandate, on hazardous chemicals found at Comprehensive Environmental Response, Compensation, and Liability Act National Priorities List sites that have the greatest public health impact. These profiles comprehensively summarize toxicological and environmental information. This article constitutes the release of portions of the Toxicological Profile for tungsten. The primary purpose of this article is to provide public health officials, physicians, toxicologists and other interested individuals and groups with an overall perspective on the toxicology of tungsten. It contains descriptions and evaluations of toxicological studies and epidemiological investigations and provides conclusions, where possible, on the relevance of toxicity and toxicokinetic data to public health. PMID- 18386526 TI - [A prognostic classification of the "floating elbow" in children]. AB - The ipsilateral fracture of the humerus and any bone of the forearm is known as "Floating elbow", being this an infrequent entity in paediatrics. Nevertheless the classification that is proposed is based specifically on clinical findings and on the radiographic appearance resulting. We performed an observational, cross-sectioned and descriptive study from January 1997 to December 2000. We included 29 patients from 4 to 14 years old male and female. All patients received treatment for "floating elbow" and they were evaluated with the application of this proposition to get their prognosis. It was made on the basis of other classifications widely demonstrated such as Holmberg's for supracondylar fractures, Kranhoft and Solgaard's for forearm fractures, Gustilo R.B. and Mendoza R.M.'s for exposed fractures and we also considered the applications of the functional classification index of Broberg and Morrey. We propose a classification system based on mechanism of injury, fracture's level, displacement, time of exposition and related injuries, with six different types, as well as the use of a prognostic classification the physician has to look for a perfect reduction of the fracture and an appropriate stabilization. PMID- 18386527 TI - [Radiographic and functional results of plate of oxnamic ram compression versus plate of compression bolted in radial and cubital fractures]. AB - OBJECTIVE: To determine if there are statistically significant differences in radiographic and functional outcome in postoperative patients with DCP (dynamic compression plate) vs. LCP (locking compression plate) in fractures radial and/or ulnar shaft. MATERIAL AND METHODS: Quasi-experimental, observational, longitudinal, comparative and prospective study, with patients surgically stabilized using the implants mentioned before with diaphyseal forearm fractures in the Balbuena General Hospital from September 1st to February 28th of 2005, with 2 years of follow-up. RESULTS: 22 patients were surgically stabilized with dynamic compression plate and 11 with locking compression plate, gender male in 88%, predominantly in 3rd life decade, earlier consolidation in the group of locking compression plate but with greater functional limitation. The differences decreased after 3 months and 6 months of clinical follow up, with no statistically significant differences at 2 years of follow-up. CONCLUSIONS: Although there are differences at the 1st and 3rd months of postoperative period, attributable to stability provided by each one of the implants, the beginning of mobilization and the soft-tissue handling, there are no statistically significant differences in the preliminary results at 6 months and finally at 2 years of follow-up. We consider that implants in study, even though of acting in a relatively different way, both of them are excellent alternatives of treatment in patients with forearm shaft fractures. PMID- 18386528 TI - [Metabolic changes and nutritional status in the spinal cord injured patient ASIA A. Evaluation and monitoring with routine laboratories, a feasible option]. AB - INTRODUCTION: Spinal Cord Injury ASIA A (tetraplegia) is a frequent pathology that may affect population regardless of age, sex or race, and that can induce metabolic abnormalities that may worsen the nutritional status of these patients. There are no existing specific protocols to treat these disorders in the specialized units in Mexico. MATERIAL AND METHODS: We analyzed 16 patients at the Spine Clinic SS-DF, 18 years or older without any drug treatment or any previous disease, known or diagnosed when admitted to the hospital. Laboratory samples were obtained as well as the nutritional status was calculated at their admittance and discharge. Individually calculated nutritional support was administered starting at their third day of hospital stay. RESULTS: 100% of the patients showed some type of metabolic disorder associated to the neurological injury. 50% of the patients were classified with severe malnutrition, 25% moderate and 25% mild, all improved at their discharge. CONCLUSIONS: We confirmed the presence of metabolic changes in these patients, the efficacy of routine laboratory in the Spine Clinic SS-DF for their evaluation, and that adequate nutritional support may help correct metabolic disorders in this patients. PMID- 18386529 TI - [Treatment of the tibial bone defects by traumatic sequels with the Ilizarov method in children]. AB - INTRODUCTION: Tibial bone defects are usually results of: high energy trauma, infections, bone tumors, and are associated to soft tissue lesion. The most successful way to fill bone defects is the use of autologous bone grafting with adequate blood supply and soft tissue coverage. PURPOSE: To evaluate the clinical and functional outcome of post-traumatic bone defects treated with vascularized bone allograft, fibular tibialization, and bone transport with Ilizarov method. MATERIAL AND METHODS: Retrospective longitudinal study with 12 patients, from October 2000 to November 2005, with a 33.6 months follow up average. RESULTS: Seven male and five female, all of them treated previously in other institutions were included in the study. We found excellent clinical and functional results in five patients, good results in five and bad in two, with an average of surgical procedures of three. COMPLICATIONS: Pseudoarthrosis, fibular fusion and non union. DISCUSSION: Postraumatic bone defects can be treated successfully using different techniques: Vascularized bone graft, tibialization, end bone transport with Ilizarov method. Bone transport allows filling of bone defect with adequate soft tissue coverage and length discrepancy management. CONCLUSIONS: These choices of treatment are different solutions with post-traumatic bone defects in patients candidate for amputation. PMID- 18386530 TI - [Arthroplasty after failed osteosynthesis in hip fractures]. AB - OBJECTIVE: To assess the functional status of patients with arthroplasty after failed osteosynthesis in hip fractures. MATERIAL AND METHODS: Prospective, descriptive, cohort of patients with primary failed osteosynthesis in hip fracture, who underwent total arthroplasty in the period 2002-2004, analyzed with the Harris functional scale. RESULTS: 26 patients: 17 women and 9 men, mean age of 74.8 years. Seventeen patients had loosening of dynamic hip screw, 5 of condylar screw 5 and 4 with cannulated screws. We made 12 hybrid, 10 cemented and 4 not cemented arthroplasties. Eighteen patients with good results, 7 with regular and 1 bad result. Patients with the highest rating were the youngest. DISCUSSION: Arthroplasty of the hip in a patient with failed osteosynthesis should be considered necessary. If we can improve pain of the hip, the patient will have less functional limitation and a better quality of life. This type of surgery requires high technical capability and experience by the orthopedic surgeon. CONCLUSIONS: Hip arthroplasty after failed osteosynthesis in hip fractures is a method that improves function and a gives a better life quality. PMID- 18386531 TI - [Review of the actions in prevention of infections in total arthroplasty of hip]. AB - The prophylactic use of antibiotics when a hip replacement procedure is performed has been subject of intense debate. Prevention of such an infection is complex, with many unresolved issues that require considerable investigation. The attentive application of reasonable principles of infection control with the goals of optimization of the wound environment, augmentation of the host response, and minimization of bacterial contamination in the preoperative, and postoperative time-period is essential to that overall reduction of the frequency of infection. We present the antibiotic prophylaxis used in the American British Cowdray Medical Center between January 1999 to July 2003. after hip arthroplasty in 532 patients. The main antibiotic used was cefalotin in 53.76% of the cases, but only in 3.2 % a one day prophylaxis treatment recommended in the literature is used, with septic loosening of 3.9%. PMID- 18386532 TI - [Treatment of the floating shoulder. Eight cases]. AB - OBJECTIVE: To show the experience in management of eight cases of floating shoulder. MATERIAL AND METHODS: Descriptive, ambispective, longitudinal and observational study of eight patients treated during the period of November 2003 to April 2005. We determined age, sex, mechanisms of injury, injuries associated to the extremity and other systems and functional results were evaluated. RESULTS: Eight patients treated, 7 men and 1 woman, age from 27 to 54 years, the average of hospital stay was of 13.25 days. The dominant extremity was injuried in the 85.5% of the cases. The most frequent morphologic pattern was a type II Ideberg glenoid fracture and a type I Allman clavicle fracture. Surgical treatment used in five patients were in the modalities of a plate to clavicle, or plate to the glenoid. Patients with acromioclavicle dislocation underwent to techniques of Phemister and Stewart. To patients treated with conservative methods were treated with an immobilizer of the shoulder plus device in 45 degrees and 90 degrees of abduction. PMID- 18386533 TI - [Freeman-Sheldon's syndrome. Report of five cases]. AB - INTRODUCTION: Freeman-Sheldon syndrome, described in 1938, is an infrequent form of arthrogryposis. MATERIAL AND METHODS: We reviewed the files of the patients with the diagnosis of Freeman-Sheldon syndrome from 1990 to 2003, and carried out an evaluation of the clinical data, family history and malformations of extremities. RESULTS: 5 patients were included, 4 male and 1 female. Upper extremities: All patients presented adducted thumbs, 3 patients were treated with a first web release and dorsal graft with improvement of grip on larger objects. Feet: Four patients had severe bilateral rigid clubfeet, they were operated on with a posteromedial release and bilateral talectomy, satisfactory results were obtained in 4 of 6 feet according to Niki criteria. Other non orthopedic anomalies: In three patients unilateral right undescended testis; 1 right inguinal hernia. CONCLUSIONS: The patients have as common factor the whistler appearance, accompanied with mill crosses hand deformity, adducted thumbs, accompanied or not with inferior extremities deformities, intellectual capacity was normal. PMID- 18386534 TI - [Sterno-clavicular plasty in anterior dislocation in child. The new surgical technic and review literature]. AB - The sternoclavicle joint takes part in stability and normal rotation of clavicle in shoulder movement. Its injury infrequent and the luxation in children is more rare. It is classified in previous and retrosternal. The most common causes are by sport trauma and car accidents. In children differential diagnosis includes proximal epiphyseal displacement of clavicle. The clinical picture is pain, sternum or thorax deformity and limitation in range of motion of the arm. The treatment is conservative or surgical, and indications to surgical treatment are pain when moving, range of motion limitation or concomitant complications. In the surgical treatment, the reduction of the clavicle is made with a percutaneous clamp and then protecting the position with a bandage in "eight". Other options are open reduction of the clavicle and subclavian plasty, reduction open and to make plastias with grafts of subclavio,fascia latae or proximal third clavicle resection and cerclage with wire. We present a clinical case and surgical treatment with a novel surgical technique. PMID- 18386535 TI - [The dilemmas of the bones neoplasms]. PMID- 18386536 TI - Chorionic villus sampling for early prenatal diagnosis at Bhumibol Adulyadej Hospital. AB - OBJECTIVE: To evaluate results of chorionic villus sampling for early prenatal diagnosis at Bhumibol Adulyadej Hospital. DESIGN: Retrospective descriptive study. SETTING: Perinatal unit, Department of Obstetrics and Gynaecology, Bhumibol Adulyadej Hospital. SUBJECTS: Three hundred and eighty three women were enrolled to chorionic villus sampling at the perinatal unit, Department of Obstetrics and Gynecology, Bhumibol Adulyadej Hospital, from November 10,1997 to October 17, 2006. RESULTS: During the present study periods three hundred and eighty three women were recruited, of these chorionic villus sampling for chromosome diagnosis were performed on 355 while 6 were for abnormal Thalassemia screening. Twenty two cases were excluded because ultrasound examination showed anembryonic pregnancy or fetal demise in utero in 13 cases, multiple fibroids in 4 cases, large area of placental hemorrhage in 3 cases, 1 case of multiple pregnancy and in 1 case the placenta was in an inappropriate position. The most common indication was elderly gravidarum (95.84%). Other indications were abnormal Thalassemia screening, abnormal ultrasound findings, family chromosome disorder previous Down syndrome, and severe oligohydramnios. The authors found eleven cases of chromosome abnormalities, four cases of maternal cell contamination and three cases of failed tissue culture (two cases from transcervical chorionic villus sampling and one case from transabdominal chorionic villus sampling) and two cases of mosaicism. There were two fetal losses in the present study and all the babies from the normal chromosome result looked normal. Second trimester amniocentesis following chorionic villus sampling was required due to maternal cell contamination, mosaicism and failed tissue culture. (2.77%) All cases had follow-up ultrasound scan during 18-20 weeks. CONCLUSION: The authors found that chorionic villus sampling is a possible alternative technique for prenatal diagnosis of cytogenetic abnormalities and abnormal Thalassemia screening in Thailand. It probably has a slightly higher rate of failed tissue culture and maternal cell contamination than amniocentesis, but it is generally done earlier in pregnancy than amniocentesis and is particularly advantageous for detecting certain genetic conditions. PMID- 18386537 TI - Effect of oral contraceptives on risk of cervical cancer. AB - OBJECTIVE: To assess the risk of oral contraceptives on the occurrence of cervical cancer. MATERIAL AND METHOD: A hospital-based case-control study was conducted. Sixty women patients with histologically confirmed invasive cervical cancer and 180 healthy women as the control group who attended the King Chulalongkorn Memorial Hospital, Bangkok, Thailand were recruited. Information about the use of oral contraceptives and other cervical cancer risk factors were obtained from personal interviews. The risk factors were evaluated by using odds ratio (OR). RESULTS: 60 women with invasive cervical cancer and 180 healthy controls were interviewed by the investigators. Compared with non-users, patients who had ever used or currently used oral contraceptive had an increased risk of cervical cancer (OR 1.45; 95% CI 0.79-2.64). However the risk was not statistically significant. Considering the duration of use, patients who had used oral contraceptives for 3 years or less did not have an increased risk of cervical cancer (OR 0.78; 95% CI 0.39-1.77). Nevertheless, the odds ratio of oral contraceptive pill use for more than 3 years was 2.57 (95% CI 1.22-5.49) which was statistically significant. CONCLUSION: Long-term use of oral contraceptive might be a cofactor that increases the risk of cervical carcinoma. Further investigations should be conducted to confirm this risk. However, Pap smear has to be done routinely in long-term oral pill users. PMID- 18386538 TI - Efficacy of etoricoxib for pain relief during endometrial biopsy; a double blind randomized controlled trial. AB - OBJECTIVE: To compare the efficacy of oral etoricoxib and placebo for pain relief during endometrial biopsy. MATERIAL AND METHOD: A double-blind, randomized controlled trial that included 80 women who underwent endometrial biopsy was done at Thammasat University Hospital between 1 September 2005 and 30 June 2006. Forty women were randomly allocated to the etoricoxib group (120 mg, tablet) and 40 to the placebo group. The main outcome was the patient's assessment of intensity of pain measured by visual analog scale (VAS) before speculum insertion, during endometrial biopsy, immediately after endometrial biopsy, and 30 minutes after endometrial biopsy. Satisfactory score was also evaluated. RESULTS: Demographic data including age, BMI, previous vaginal deliveries, previous pelvic surgery and history of curettage were not significantly different between the etoricoxib group and the placebo group. Mean pain score in the etoricoxib group was not significantly lower when compared with the placebo group during endometrial biopsy (5.0 +/- 1.7 versus 5.25 +/- 2.2, p = 0.7) and immediately after endometrial biopsy (2.1 +/- 2.2 versus 2.8 +/- 1.7, p = 0.1) but significantly lower at 30 minutes after endometrial biopsy (0.2 +/- 0.5 versus 0.6 +/- 0.8, p = 0.01). Mean satisfactory score was significantly higher in the etoricoxib group (6.9 +/- 1.8 versus 5.1 +/- 2.3, p = 0.001). CONCLUSION: A single oral dose of etoricoxib for reduction of pain during endometrial biopsy had not significantly lower the pain score during the procedure compared with the placebo. However mean satisfactory score in the etoricoxib group was higher with statistically significant difference. Also the authors found no serious adverse effects of this drug. PMID- 18386539 TI - Efficacy of external cephalic version with tocolysis in late pregnancy. AB - OBJECTIVE: To study the success rate and identify factors influencing the success rate of external cephalic. version (ECV) at Bhumibol Adulyadej Hospital. STUDY DESIGN: Prospective descriptive study. SETTING: Department of Obstetrics and Gynecology, Bhumibol Adulyadej Hospital. MATERIAL AND METHOD: All parturients attending the obstetrics unit at Bhumibol Adulyadej Hospital between October 1, 1997 and September 30, 2006, having completed 36 or more gestational weeks with singleton non-vertex fetus, who had no exclusion criteria for ECVwere given full information concerning a trial of ECV risk of cesarean section, and risk of emergency breech deliveries. Those who chose to undergo ECV after counseling were recruited and gave signed consent. One hundred and forty singleton, pregnant women with non-vertex presentation participated in this study. RESULTS: The success rate of ECV was 71.43%. Birth weight significantly affected the success rate of ECV Maternal weight, parity, gestational age, and placental site did not have any effect on the outcome. All fetuses in the present study were subsequently delivered without significant morbidity and no cases of perinatal mortality were recorded. CONCLUSION: ECV is a safe procedure with a high success rate in selective cases. ECV thus, is an effective alternative practice for non vertex presentation, which can also reduce the rate of breech delivery and cesarean section. The major benefits of external cephalic version are reduced maternal morbidity and mortality from surgery. PMID- 18386540 TI - Perinatal outcomes in severe preeclamptic women between 24-33(+6) weeks' gestation. AB - OBJECTIVE: The present study was undertaken to determine perinatal and maternal outcomes in severe preeclamptic women between 24-33-week gestation and compare the outcomes between expectant and aggressive management. MATERIAL AND METHOD: A retrospective descriptive study of 99 women with severe preeclampsia between 24 33(+6)-week gestation who delivered at the King Chulalongkorn Memorial Hospital from January 2002 to December 2005 was included. The outcomes were analyzed according to the gestational age at the time of admission (< 28 weeks' and > or = 28 weeks' gestation) according to expectant compared with aggressive management. Statistical analysis was performed by student t-test and chi2-test. RESULTS: The perinatal morbidity and mortality were significantly high in the gestational age < 28 weeks group. There were 11 perinatal deaths, 8 in those managed at < 28 weeks and 3 in those managed at 28-29 weeks' gestation (p < 0.05). Maternal morbidities were similar among both groups. There was no maternal death. CONCLUSION: Delivery at remote from term, especially < 2-week gestation, increases neonatal morbidity and mortality in severe preeclamptic women. Expectant management should be done in the tertiary care center with close maternal and fetal monitering. PMID- 18386541 TI - Treatment failure following large loop excision of the transformation zone for the treatment of cervical intraepithelial neoplasia at Rajavithi Hospital. AB - OBJECTIVE: To evaluate the treatment failure rate of large loop excision of the transformation zone (LLETZ) in the treatment of cervical intraepithelial neoplasia (CIN) and the risk of persistence or recurrence depending on the clinicopathologic factors. MATERIAL AND METHOD: Retrospective case-record review of 250 women who had a final diagnosis of CIN and underwent LLETZ in Rajavithi Hospitalfrom June 1st, 1998 to December 31st, 2003. Computerized files of these patients were then reviewed for clinicopathological follow-up results. RESULTS: Success rate of 86.8% was obtained. The incidence of treatment failure was 13.2%. The clinicopathologic factor associated with the persistence or recurrence was the presence of CIN at the margin of excision. Of 69 cases with incomplete excision (positive margin), treatment failure developed in 29.0%, compared to 5.2% in patients with complete excision (negative margins) (p < 0.001). Using multivariate analysis, incomplete excision and endocervical margin involvement of specimen were independent risk factors for the treatment failure of CIN. CONCLUSION: LLETZ is an effective treatment for CIN. Treatment failure rate is low. Positive surgical margin is a predictor of persistence or recurrence after LLETZ. Incomplete excision and endocervical margin involvement of specimen are significant independent risk factors. PMID- 18386542 TI - A study of Clostridium difficile-associated disease at King Chulalongkorn Memorial Hospital, Thailand. AB - BACKGROUND: Currently, in established antibiotic era, there is a widespread and increasing use of broad-spectrum antibiotics. Clostridium difficile, one of the troublesome intruders, flourishes when normal gut flora is altered by antibiotics. C. difficile is recognized as a frequent and leading cause of antibiotic-associated diarrhea and colitis. It causes substantial morbidity and mortality in hospitalized patients. OBJECTIVE: The present study was aimed at determining patient characteristics, clinical features, treatment, and outcomes of C. difficile-associated disease (CDAD) in hospitalized patients at King Chulalongkorn Memorial Hospital, Bangkok, Thailand. MATERIAL AND METHOD: From 2002 to 2005, 88 patients with positive latex immunoassay for C. difficile toxin A were identified. Data from medical records of 56 patients were available for analysis. RESULTS: Of 56 patients, there were 28 males and 28 females, with the mean age of 47.39 years (range: 4 months to 93 years). 50 (89.3%) patients had underlying illnesses with hematological malignancies (14 patients, 25%) and solid tumors (15 patients, 26.8%) being the most common. All patients had a history of antibiotic use including current (17 patients, 30.4%), recent (16 patients, 28.6%), or both current and recent uses (23 patients, 41.1%). Cephalosporins and carbapenems were the two most commonly prescribed antibiotics. 25 (44.6%) patients were receiving either omeprazole or ranitidine. 12 (21.4%) patients had received chemotherapy within two months before CDAD diagnosis. Of 50 stool specimens examined, only 26 (52%) had white or red blood cells. Colonoscopy was performed in only three patients, and pathological findings revealed non-specific colitis. Oral metronidazole, intravenous metronidazole, and vancomycin were prescribed for CDAD treatment in 38 (67.9%), 4 (7.1%), and 2 (3.6%) patients, respectively. 8 (14.3%) patients had no specific treatment, and the offending antibiotic was not discontinued in three of them. An overall initial response rate was 66.7%. 2 patients relapsed after metronidazole treatment. CONCLUSION: The present study is the first in Southeast Asia to describe the decreased initial response rate of metronidazole treatment of CDAD. The reasons for this relatively poor response in the presented patients need to be determined in a future study. PMID- 18386543 TI - Predictive factors of systemic embolism in patients with mitral stenosis in sinus rhythm. AB - BACKGROUND: Systemic embolism is one of the major complications in patients with mitral stenosis (MS) who are in atrial fibrillation; however, this serious complication can also occur in patients with MS in sinus rhythm. OBJECTIVE: The purpose of the present study was to identify the predictive factors of systemic emboli in patients with MS in sinus rhythm. MATERIAL AND METHOD: Twenty patients with MS in sinus rhythm with recent cerebral embolism and 32 with MS in sinus rhythm without any history of systemic embolism were studied between January 2004 and May 2006. Clinical and echocardiographic data were assessed using stepwise logistic regression for prediction of systemic embolism. RESULTS: Age (odds ratio [OR], 1.14; 95% confidence interval [CI] 1.04-1.26) and left atrial spontaneous echo contrast (LASEC) grade 3+ and 4+ (OR, 46.42; 95% CI, 5.00-436.49) were associated with, and predictive of systemic emboli, whereas left atrial appendage contraction flow velocities, left atrial size and mitral valve area were not. CONCLUSION: The present study demonstrates that age and LASEC are the major predictive factors of systemic embolism in patients with MS in sinus rhythm. It would therefore be prudent to give anticoagulants to patients in atrial fibrillation as well as to those in sinus rhythm at risk of systemic embolism. PMID- 18386544 TI - Effectiveness of double-surface intensive phototherapy versus single-surface intensive phototherapy for neonatal hyperbilirubinemia. AB - OBJECTIVE: To compare the efficacy and adverse effects of double-surface intensive phototherapy (DsIPT) and single-surface intensive phototherapy (SsIPT) in term newborn infants with hyperbilirubinemia. DESIGN: Prospective randomized controlled trial. MATERIAL AND METHOD: Healthy full-term infants with nonhemolytic hyperbilirubinemia (total serum bilirubin between 13.0 to 19.9 mg/dl) were allocated randomly to two modes of phototherapy. Group 1 infants received single-surface intensive phototherapy. Group 2 infants received double surface intensive phototherapy Serum bilirubin, body weight, body temperature and number of defecation were measured at 24 and 48 hours after phototherapy. RESULTS: Sixty infants were studied, 30 in each group. Baseline characteristics were similar in both groups. The initial mean serum bilirubin had no statistically significant difference between SsIPT and the DsIPT groups. After 24 and 48 hours of phototherapy, mean serum bilirubin in the group receiving SsIPT declined 3.5 +/- 1.7 and 6.5 +/- 2.3 mg/dl, while in the DsIPT group, the mean serum bilirubin declined 5.4 +/- 2.0 and 8.4 +/- 2.1 mg/dl respectively. The mean body temperature after 24 hours of phototherapy in the DsIPT group was higher than SsIPT group significantly. The number of defecation in the SsIPT group increased significantly after 24 and 48 hours of phototherapy, but the body weight change in both groups was not statistically significant. CONCLUSION: DsIPT was significantly more effective in reducing serum bilirubin than SsIPT in the term jaundiced infants after 24 and 48 hours of treatment. The adverse effect of DsIPT found in the present study was the increased of body temperature after 24 hours of phototherapy. PMID- 18386545 TI - Thyroid functions in children with Down's syndrome. AB - OBJECTIVE: To evaluate thyroid function in children with Down's syndrome, and to ascertain the presence of a relationship between overt thyroid diseases and congenital anomalies. MATERIAL AND METHOD: One hundred and forty Down's syndrome patients, aged from 3 days to 13 years 9 months, were evaluated for karyotype, thyroid functions and the coexistence of congenital anomalies. RESULTS: Trisomy 21 was found in the majority of cases (95.7%). Fifty-six patients (40%) had abnormal thyroid functions: 53 (37.9%) hypothyroidism and 3 (2.1%) hyperthyroidism. Ten patients (7.1%) were diagnosed with overt thyroid disease: congenital hypothyroidism 3.6%, acquired hypothyroidism associated autoimmune thyroiditis 1.4% and hyperthyroidism 2.1%. None of the patients with congenital hypothyroidism had athyreosis or ectopic thyroid gland. Sub-clinical hypothyroidism accounted for 32.9% of all cases; 10.7% showed a spontaneous decrease to normal TSH levels and 13.6% had persistently elevated TSH levels with the median follow-up time of 6 and 12 months, respectively. Congenital heart disease, gastrointestinal anomalies and hematological disease were found in 73.6, 10 and 3.6 percent of patients, respectively. There was no statistical correlation between the coexistence of cardiovascular or gastrointestinal disease in Down's syndrome patients with overt thyroid diseases or sub-clinical hypothyroidism to those having normal thyroid functions. CONCLUSION: Sub-clinical hypothyroidism was the most common thyroid abnormality in children with Down's syndrome. A longitudinal and timely-scheduled evaluation of thyroid function is needed to establish the natural course of this abnormality and the proper management guideline. PMID- 18386546 TI - Comparison of caulking gun and standard cement gun using for femoral cementation. AB - BACKGROUND: Currently the standard femoral cementing technique requires cement injection gun to deliver bone cement into the femoral canal. The standard cement gun is expensive and must be imported. There is no previous study about the use of a household caulking gun for femoral cementation. OBJECTIVE: To compare the radiographic quality of cementing technique in proximal femur between using standard cement gun and caulking gun. MATERIAL AND METHOD: Experimental study was performed on ten pairs of adult bovine femora. After the proximal femoral canal was prepared for cementation, each pair of bovine femora was randomly selected to be injected with bone cement with a standard cement gun in one side. The other side was injected with a caulking gun and the authors' invented kit. The prosthesis was inserted and radiographs were taken. Radiographic quality of cement interdigitation was evaluated by the modified Barrack's cement grading score. Cement distribution was categorized into 14 zones of Gruen. The data was statistically analyzed by Wilcoxon matched pairs signed-rank test. The inter observer and intra-observer agreement was analyzed by Kappa analysis. RESULTS: Radiographic quality score of femoral cementation in the group using the caulking gun was not statistically different from in the group using the standard gun in all 14 Gruen zones (p > 0.05). Intra-observer and inter-observer agreement were moderate (Kappa = 0.71 and 0.59 respectively). CONCLUSION: The radiographic quality of femoral cementation using the caulking gun was not significantly different from the standard cement gun. The caulking gun and invented kit could be applied in clinical use and save the operative cost. PMID- 18386547 TI - Prevention of post operative pain after abdominal hysterectomy by single dose etoricoxib. AB - OBJECTIVE: To test whether a reduction in post operative morphine consumption could be achieved by a single-dose of etoricoxib before induction of anesthesia. DESIGN: Randomized, double-blind, placebo-controlled study. MATERIAL AND METHOD: Two hours before surgery, patients undergoing transabdominal hysterectomy (under general anesthesia) were randomized to a single oral dose of 1) etoricoxib 120 mg (n = 17), 2) etoricoxib 180 mg (n = 17), or 3) placebo (n = 15). Intravenous morphine was given for patient-controlled analgesia (PCA) device. Morphine consumption, pain scores both at rest and on coughing, and side-effects were recorded at 1, 2, 4, 8 and 24 h after surgery. Patients' global evaluation of study medication was assessed at the end of the present study. RESULTS: Etoricoxib provided greater clinical benefit than the placebo in terms of mean morphine in milligram at 24 hour consumption (stardard deviation): a) 26.4 mg (SD of 11.2) for etoricoxib 120 mg; b) 27.2 mg (SD of 9.9) for etoricoxib 180 mg; and, c) 36.6 mg (SD of 8.9) for the placebo group. At 8 h post surgery, pain both at rest and on coughing in the active drug groups was significantly less than in the placebo, while pain on coughing was significantly less at 24 h. Patients reported better global satisfaction and less somnolence in the etoricoxib groups. CONCLUSION: Single dose etoricoxib 180 mg given before surgery provides the same analgesic effect as 120 mg for post operative pain after an abdominal hysterectomy. PMID- 18386548 TI - Benign and malignant soft tissue mass: magnetic resonance imaging criteria for discrimination. AB - OBJECTIVE: Determine MRI features to differentiate between benign and malignant lesions. MATERIAL AND METHOD: The Magnetic resonance imaging (MRI) taken in the 5 year period (2003-2007) of 85 patients with benign and malignant soft tissue masses were analyzed. The criteria for discrimination were based on size, origin, signal homogeneity on T1- and T2-weighted, perilesional edema or invasion, hemorrhage, necrosis, and bone and neurovascular involvements. RESULT: There were 50 benign and 35 malignant lesions. Eighty-two lesions had histologic proof while three lesions had MRI characteristics of benign lipoma. CONCLUSION: No single MRI feature was diagnostic for any soft tissue tumors in the present study. However, three individual MRI features were statistically significant for differentiation between benign and malignant soft tissue masses (p < 0.05). The features that favor malignancy were heterogeneous signal on T2-weighted, perilesional edema or invasion, and necrosis in the masses. PMID- 18386549 TI - Perspective automated inkless fingerprinting imaging software for fingerprint research. AB - Fingerprint collection using ink-and-paper image is a conventional method i.e. an ink-print, transparent-adhesive tape techniques which are slower and cumbersome. This is a pilot research for software development aimed at imaging an automated, inkless fingerprint using a fingerprint sensor, a development kit of the IT WORKS Company Limited, PC camera, and printer The development of software was performed to connect with the fingerprint sensor for collection of fingerprint images and recorded into a hard disk. It was also developed to connect with the PC camera for recording a face image of persons' fingerprints or identification card images. These images had been appropriately arranged in a PDF file prior to printing. This software is able to scan ten fingerprints and store high-quality electronics fingertip images with rapid, large, and clear images without dirt of ink or carbon. This fingerprint technology is helpful in a potential application in public health and clinical medicine research. PMID- 18386550 TI - Analysis of polymorphisms in the 5'-flanking region of the apolipoprotein(a) [Apo(a)] gene in Thai subjects with coronary artery diseases. AB - Lipoprotein(a) [Lp(a)] is a complex lipoprotein particle in human plasma. It is composed of apolipoprotein B (Apo B)-100 and apolipoprotein(a) which are linked by a disulfide bond. Plasma levels of the Lp(a) vary greatly (over 1,000 folds) among individuals. Elevated plasma levels of the Lp(a) have been shown to be an independent risk factor for coronary artery diseases (CAD). The level of Lp(a) is controlled by a single gene, the Apo(a) gene, with multiple alleles; each encodes different concentrations of the Lp(a). Previous studies revealed the presence of polymorphisms in the 5'-flanking region (FL) of the Apo(a) gene at 3 positions: G or A (-914), C or T(-49), and G or A (-21), which can be detected by cleavage of PCR-amplified DNA products with TaqI, MaeII and HhaI, respectively. The 5'-FL genotypes of the Apo(a) gene can be classified by the combination of the presence (+) or absence (-) of these restriction sites into 5 types; type A, +++, type B, ++, type C, -+-, type D, --+ and type E, +-+. In the present study, the authors analyzed the 5' FL types of the Apo(a) gene by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) in 100 healthy control subjects, 26 CAD patients with [Lp(a)] < or = 30 mg/dL, and 94 CAD patients with [Lp(a)] > 30 mg/dL. The authors found that the genotype frequencies of the Apo(a) gene were 53, 16, 27 and 4%, for types A, B, C and D respectively in normal healthy controls. In CAD patients with [Lp(a)] < or = 30 mg/dL, the distribution of the genotype frequencies were 53.8, 11.5, 30.8 and 3.9% for types A, B, C and D, respectively. Additionally in CAD patients with [Lp(a)] > 30 mg/dL, the genotype frequencies were 60.6, 11.7, 21.3 and 6.4% for types A, B, C and D, respectively. The present study might shed some light to understand CAD at the molecular level. PMID- 18386551 TI - Hypoglycemic effect of sitting breathing meditation exercise on type 2 diabetes at Wat Khae Nok Primary Health Center in Nonthaburi province. AB - OBJECTIVE: To evaluate the hypoglycemic effect of Somporn Kantaradusdi Triamchaisri technique 1 (SKT1) of sitting breathing meditation exercise on type 2 diabetic patients. MATERIAL AND METHOD: This quasi experiment study was performed on type 2 diabetic patients at Wat Khae Nok primary health center from April to May 2007 for a two-week period every Tuesday of the week (3 visits). At the first visit, the patients were educated about diabetes self care after breakfast. At the second and third visit, the participants were trained to practice SKT1 as intervention after breakfast. Post prandial plasma glucose and blood pressure before and after the intervention were recorded. Descriptive statistics and paired t-test were used to analyze the study data. RESULTS: Fifty patients participated in the present study, 11 were males and 39 females. They had a mean age of 63.1 years (range 42-80). The present study revealed that SKT1 significantly reduced levels of postprandial plasma glucose by 19.26 +/- 30.99 mg/dl, (p < 0.001) in the second visit, and 17.64 +/- 25.48 mg/dl, (p < 0.001) in third week visit. Furthermore, systolic blood pressure in the second visit was significantly decreased by 6.49 +/- 11.77 mmHg, (p < 0.001). Diastolic blood pressure in the third visit was also significantly decreased by 3.04 +/- 9.79 mmHg, (p < 0.05). CONCLUSION: The present study showed that SKT1 practice in type 2 diabetic patients had a post prandial hypoglycemic effect and a slight reduction to systolic and diastolic blood pressure. PMID- 18386552 TI - Utilize the modified Delphi technique to develop trauma care indicators. AB - OBJECTIVE: Develop performance indicators reflecting the quality of trauma patient-care in the emergency room and hospitalization within the first 48 hours. MATERIAL AND METHOD: A two-round Delphi technique was conducted. A panel of 11 expert surgeons experienced in the fields of trauma care was consulted. The panel was initially asked to rate performance indicators that reflected the quality of trauma care given in the emergency department setting and hospitalization in the first 48 hours using a 5-point on visual analogue scale. The statement of indicators that was collected from the first round was analyzed and necessary changes were before resending to the same experts. Each indicator statement was considered consensus if the expert's opinion rating was 4 or 5 for more than 70% (8 out of 11 experts). RESULTS: Fifty-three performance indicators were proposed in four domains of trauma care in emergency room and four domains of trauma care in hospitalization within 48 hours. Altogether 35 indicators reached consensus reflecting quality of trauma performance after two rounds. Twenty-one of these were trauma care indicators in emergency room and 14 were trauma care indicators in hospitalization within 48 hours. CONCLUSION: Twenty-one indicators of quality of trauma care in the emergency room and 14 in the hospitalization within 48 hours have been developed. They will be used as the tool by specialist for quality evaluation in the next phase. PMID- 18386553 TI - Hemophagocytic syndrome in Dengue hemorrhagic fever with severe multiorgan complications. AB - A 46 year old woman who presented with severe multiorgans involvement including liver brain, cardio-pulmonary failure, gastrointestinal bleeding, progressive cytopenia, DIC and hemophagocytic syndrome during the convalescent phase of Dengue type II has been successfully treated primarily with pulse methyl prednisolone and high dose intravenous immunoglobulin G. The authors believe that HPCS are not infrequently seen with high mortality and recommended early diagnosis and treatment with the regimen. This is the first complete report of hemophagocytic syndrome in adult dengue hemorrhagic fever in Thailand. The literature of HPCS in DHF was reviewed and discussed. PMID- 18386554 TI - Maternal chickenpox in peripartum period: a case report and review. AB - A 30-year-old pregnant woman had skin lesions at 38 weeks of gestation. She was diagnosed as primary varicella zoster infection. Her clinical symptoms were high fever and generalized vesicles eruption. No serious maternal complication was found. The patient delivered a male baby 4 days after she developed skin lesions. The neonatal blood IgM against varicella zoster was negative. The baby was given Varicella zoster immunoglobulin within 24 hours and isolated in a neonatal care unit. The baby developed skin lesions on day 11th post delivery. Intravenous acyclovir was administered for 7 days. Finally, the baby was found to be free of severe neonatal varicella infection. PMID- 18386555 TI - Necrotizing skin and soft-tissue infections associated with septicemia: 7 cases report and review. AB - The authors report seven cases of necrotizing skin and soft-tissue infections, with clinical presenting as hemorrhagic bullae, gangrenous cellulitis or necrotizing fasciitis, in association with septicemia, between January 2003 and January 2007 in Hat Yai Hospital. Six were male and the majority of the lesions, six cases, occurred in the lower extremities. The average age of the patients was 50.0 +/- 11.019 years old. All patients presented with watery diarrhea, severe abdominal pain, high fever and sepsis. The skin lesions were begun with erythema, tender and swelling with formation of hemorrhagic bullae, gangrene and necrosis within 24-48 hours. Three of them were caused by Streptococcus spp., another three by Halophilic Vibrios, and only one by Aeromonas hydrophila. Furthermore, the literatures related with clinical manifestations of necrotizing skin and soft tissue infections, etiologic pathogens, histological finding, management in setting of sepsis, comorbid conditions, complications and patients' outcome were reviewed. PMID- 18386556 TI - Policy roadmap for stem cell technology in Thailand. AB - Policy and technology roadmaps have been long and widely used in industry and business sectors. The primary objective of the roadmap is to be a policy and technology planning tool helping to deal with an increasingly competitive environment. The obvious benefit of roadmapping is to provide information to make better technology investment decisions by identifying critical technologies and technology gaps and identifying methods to improve research and development (R&D) investments. It can also be used as a marketing tool. Roadmapping is critical and necessary when the technology investment decision is not straightforward. This occurs when it is not clear which alternative to pursue, how soon the technology is needed, or when there is a need to coordinate the development of multiple technologies. Stem cell technology is still in its nascent stage and one of the technologies with obvious uncertainties. Moreover it involves many issues from bioethical, legal and public policy perspectives. Then, development of national policy and technology roadmap for stem cell technology is definitely required and crucial to make most benefit from this promising technology for Thailand. The present article will provide perspectives on stem cell policy roadmap and propose critical action plans for the next five-year period. PMID- 18386557 TI - Ethics and laws. PMID- 18386558 TI - Start the new year with proper oral health care. PMID- 18386560 TI - Hyperglycemia and sick day management in students with diabetes. PMID- 18386561 TI - Salaries rise, but at a slower rate. PMID- 18386562 TI - The American school nurse: expanding multicultural knowledge. Part 1. PMID- 18386563 TI - The hardest job you'll ever love. PMID- 18386564 TI - MyPyramid: a review. PMID- 18386565 TI - Teen fathers: an introduction. PMID- 18386566 TI - Understanding sudden loss and traumatic grief at school. PMID- 18386567 TI - [Effect of rhubarb combined with enteral nutrition on C-reactive protein and interleukin-6 in peripheral blood of patients with gastric cancer during perioperative period]. AB - OBJECTIVE: To observe the effects of rhubarb and enteral nutrition on C-reactive protein (CRP) and interleukin-6 (IL-6) in peripheral blood of patients with gastric cancer during perioperative period. METHODS: Fifty-six patients with gastric cancer were randomly assigned to three groups. The 20 patients in Group A were given conventional fluid supplement and parenteral nutrition; the 21 patients in Group B were given conventional fluid supplement and enteral nutrition via nasal feeding starting from the 42nd hour to the 8th day after operation; and the 15 patients in Group C were treated, besides the same fluid supplement and enteral nutrition as that for Group B, with additional three times of 10% rhubarb liquid medication, given at the day before operation (100 mL), and the 18th and 42th hour post-operation (50 mL in each time). Serum levels of CRP and IL-6 of all patients were measured at different time points, i. e. the day before operation (T0), the 1st (T1), 3rd (T2) and 7th day (T3) after operation. RESULTS: Acute inflammatory reaction happened in all the patients. However, the plasma level of CRP and IL-6 in Group C at T3 (12.67 +/- 4.28 mg/L and 18.17 +/- 11.10 mg/L) was significantly lower than those in Group A (19.72 +/- 9.73 mg/L and 20.44 +/- 6.32 mg/L) and Group B (21.27 +/- 10.46 mg/L and 20.29 +/- 7.79 mg/L), respectively (P < 0.05). And the recovery time of borborgmus in Group C were the shortest among the three groups. CONCLUSION: Applying rhubarb on gastric cancer patients during perioperative period can effectively decrease the levels of CRP and IL-6 in peripheral blood, alleviate acute inflammatory reaction caused by operative trauma, promote the recovery of postoperative gastrointestinal motility and be beneficial for the supplement of enteral nutrition. PMID- 18386568 TI - [Effect of traditional Chinese medicine on quality of life and survival period in patients with progressive gastric cancer]. AB - OBJECTIVE: To investigate the effect of traditional Chinese medicine (TCM) on quality of life (QOF) and survival period in patients with progressive gastric cancer, and thus exploring its clinical efficacy. METHODS: TCM therapy applied in the 34 patients assigned in the TCM group (Group I ) included intravenous injection of Cinobufotalin, beta-elemene, or orally taking of anti-cancer Chinese herbs. The same TCM was also applied in the 36 of the combined treatment group (Group II), but in combined use of FOLFOX chemotherapeutic protocol. Twenty-one days was taken as one cycle and all the patients received 2 cycles of treatment. RESULTS: The median survival period in group II was 31 months, while it was 30 months in group I; the 1-, 2-, 3-year survival rates in group II were 88.89%, 84.38% and 59.26%, and those in the group I were 82.35%, 71.43% and 65.00%, respectively with insignificant difference between the two groups (chi2 = 0.298, P > 0.05); QOF in group I was significantly superior to that in group II (P < 0.05), and the adverse reaction occurrence was significantly less in Group I than that in group II. CONCLUSION: Chinese medicine treatment can improve the QOF and prolong the survival period of patients with progressive gastric cancer with few side effects. PMID- 18386569 TI - [Effect of local application of allicinvia gastroscopy on cell proliferation and apoptosis of progressive gastric carcinoma]. AB - OBJECTIVE: To study the effects of local application of allicin via gastroscopy on progressive gastric carcinoma, and to investigate its possible mechanisms. METHODS: Eighty patients with progressive gastric adenocarcinoma, whose diagnosis was confirmed by gastroscopy and pathological examination, were assigned to 2 groups, 40 in each group. Forty-eight hours before operation, allicin was infused via gastroscopy to the lesion region of patients in the allicin group, and normal saline was infused instead to those in the control group. The gastric carcinoma tissue gotten from gastrectomy was taken to determine the percentage of cells in various cell cycle phases ( G0/ G1, S and G2/M), the cell apoptosis rate, proliferation index value and apoptosis related gene protein such as Fas, Bax and Bcl-2 by flow cytometry. RESULTS: In the allicin group, the cell apoptosis rate was 9.60 +/- 1.52%, the percentage of cell in G0/G1 phase was 72.12 +/- 8.35%, in G2/M phase 9.54 +/- 3.20%, and PI 27.80 +/- 8.35, while in the control group, the corresponding data was 2.20 +/- 0.58%, 69.56 +/- 5.15%, 13.20 +/- 3.05%, and 30.40 +/- 5.15, respectively, and significant difference in all the 4 indexes could be found between the two groups (P < 0.05, P < 0.01). Moreover, allicin showed effects in up-regulating the protein expressions of apoptosis promoting gene Bax and apoptosis initiating gene Fas (P < 0.05, P < 0.01), and down regulating that of anti-apoptosis gene Bcl-2 (P < 0.05). CONCLUSION: Local application of allicin via gastroscopy can inhibit the cell growth and proliferation of progressive gastric carcinoma, and can also promote gastric carcinoma cell apoptosis. PMID- 18386570 TI - [Randomized controlled study on effect of Quxie capsule on the median survival time and qualify of life in patients with advanced colorectal carcinoma]. AB - OBJECTIVE: To evaluate the effect of Quxie Capsule (QXC) on the median survival time and quality of life in patients with advanced colorectal carcinoma. METHODS: Forty patients with advanced colorectal carcinoma were observed in a randomized controlled clinical trial (RCT). Out of them the 37 evaluable patients were assigned into the treatment group (18 patients) and the control group (19 patients). They were all treated by the routine treatment for cancer, including chemotherapy and radiotherapy, while those in the treatment group were administered with QXC additionally. After being treated for 3 months, the follow up study was carried out to evaluate the fatality rate (FR), survival time (ST), median survival time (mST), time to progression (TTP), and quality of life (QOF). RESULTS: FR, mST, ST and TTP in the treatment group were 11.1% (2/18), 17 months, 22.63 +/- 7.34 months, and 17.76 +/- 5.62 months respectively, and those in the control group were 42.1% (8/19), 13 months, 19.76 +/- 8.28 months and 12.68 +/- 9.26 months respectively, showing significant difference between the two groups (P < 0.05). The scores of TCM symptom, QOF and KPS in the treatment group were 15.59 +/- 3.78, 54.06 +/- 3.96 and 64.71 +/- 6.24 before treatment, and 10.53 +/- 5.57, 58.65 +/- 4.03, 69.41 +/- 4.29 after treatment, respectively, showing significant improvement (P < 0.05). While the three scores in the control group were 16.11 +/- 3.99, 54.06 +/- 4.39 and 64.44 +/- 5.11 before treatment, and 19.61 +/- 7.78, 50.17 +/- 8.26 and 60.00 +/- 9.70 after treatment, respectively, showing a statistically significant worsening tendency in the latter two (P <0.05). CONCLUSION: QXC can reduce the FR, prolong the ST, mST, mCFPT, and improve the QOF in patients with advanced colorectal carcinoma. PMID- 18386571 TI - [Exploration into relationship between TCM syndrome type and laboratory indexes in patients with systemic lupus erythematosus]. AB - OBJECTIVE: To investigate the relationship between TCM syndrome type and laboratory indexes in patients with systemic lupus erythematosus (SLE). METHODS: Three hundred and eighty-three SLE patients were differentiated into six syndrome types based on the "toxin" sydrome differentiation of TCM, Type 1, the heat-toxin flourishing type; Type 2, the stasis heat with toxin type; Type 3, the turbid toxin congested type; Type 4, the general deficiency with evil stay type; Type 5, the heat-toxin burning yin type; Type 6, the Gan-stagnant with toxin convergency type. The indexes, including complement C3 (C3), immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), alanine aminotransferase (ALT), 24 h urinary protein quantitation, white blood cell (WBC) and platelet (PLT) count, were determined, and the SLEDAI score was calculated. RESULTS: Levels of IgA and IgM in all patients were basically normal. Level of IgG was higher than the normal range in patients of type 1 and 2, being 18,713.81 mg/L and 23,131.54 mg/L respectively, showing significant difference between the Type 2 and Type 3 (P < 0.05); the lowest mean value of C3 presented in patients of type 1 (586.32 mg/L), and that in patients of type 4 was significantly different to that in patients of other types (P < 0.05); count of WBC and PLT was lower in patients of type 5 and significantly different to that in the other five types (P < 0.05); level of 24 h urinary protein quantitation was higher than normal in all the patients and the highest level (2.78 g/24 h) was found in patients of type 3; and the highest level of ALT (112.75 U/L) appeared in patients of type 6 as compared with that in patients of other types. CONCLUSION: Different variations of laboratory indexes present in SLE patients of different syndrome types, there are certain relationship between the TCM syndrome types and the laboratory indexes, and these rules may provide reference for evaluating clinical therapeutic effects. PMID- 18386572 TI - [Effect of Ningzhi capsule on blood lipid spectrum in type 2 diabetes mellitus patients complicated with hyperlipidemia]. AB - OBJECTIVE: To observe the effect of Ningzhi Capsule (NZC) on blood lipid spectrum in type 2 diabetes mellitus patients complicated with hyperlipemia (DM-HL). METHODS: Adopting randomized, parallel and controlled trail method, a total of 70 DM-HL patients of qi-yin deficiency and phlegm-blood stagnant syndrome type were randomized into two groups. The original medication for lowering blood sugar and blood pressure was unchanged, the trial group received oral administration of NZC 5 tablets, 3 times a day, while the control group received Lipanthgl or Simvastatin depending on their different constituents of blood lipids. After 6 months of treatment, sixty subjects completed the trail while two patients dropped out due to side effect and 8 patients lost follow-up (4 in each group). Levels of blood lipids, blood routine, liver and kidney function and symptoms in patients were detected and compared. RESULTS: After treatment, levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL-C), apoprotein B, and lipoprotein a (LPa) lowered, while levels of high-density lipoprotein (HDL C) and apoprotein A raised in the trial group as compared with those before treatment (P < 0.05, P < 0.01), but showed no difference between the two groups after treatment except HDL level (P > 0.05). Scores of symptoms were also lowered significantly in the trial group (P < 0.01). In the observation period, no abnormal findings in blood and urine routine examination as well as in liver and renal function were found. CONCLUSION: NZC could lower the blood lipid spectrum and improve the TCM symptoms in DM-HL patients without any adverse reaction. PMID- 18386573 TI - [Study on fMRI brain map in patients undergoing needling at Zusanli (ST36) by reinforcing method]. AB - OBJECTIVE: To observe the characteristic of the fMRI brain map in patients undergoing needling at Zusanli (ST36) by reinforcing method for exploring the essence of Meridian-Collaterals and the mechanisms of acupuncture in treating diseases. METHODS: Twenty-six healthy volunteers were randomly assigned to two groups by double blinded method, 16 in the acupoint group and 10 in the non acupoint group. Using GE Signa 1. 5 T superconducting MRI system, the fMRI was performed with Gradient echo-EPI sequence. Post-processing of fMRI data was performed using the Functool software (GE-ADW4.0) to generate positive correlation coefficient brain functional activating images and the data was analyzed statistically using SPSS 13.0 software. RESULTS: Brain functional area was elicited in 13 out of the 15 patients in the acupoint group and 10 in the non acupoint group. Among them, the temporal elicited area in the acupoint group showed specificity (Fisher's Exact test, P = 0.022) and only the difference in contralateral hemisphere activation rate was of statistical significance (McNemer test, P = 0.020). CONCLUSION: Acupoints has its own specific brain activated areas. The therapeutic effect of acupoint might be mediated through brain to treat diseases and regulate functional disorder of organs. There exists special transmission channel of meridian. PMID- 18386574 TI - [Effects of some active ingredients of Chinese drugs for activating blood circulation and detoxicating on blood lipids and atherosclerotic plaque inflammatory reaction in ApoE-gene knockout mice]. AB - OBJECTIVE: To observe the effects of active ingredients from Chinese drugs for activating blood circulation and detoxicating, including notoginseng saponins (drug 1), Coptis chinensis (drug 2), giant knotweed rhizome (drug 3) and rhubarb (drug 4), on blood lipids and inflammatory reaction of aortic atherosclerotic plaques in ApoE knockout mice. METHODS: ApoE knockout mice were fed with high-fat diet for 26 weeks, then they were randomized into 6 groups, the untreated model group and the test groups treated with various test drugs respectively. After ending the 13 weeks of treatment, all the mice were sacrificed with their blood lipids detected, and their heart and aorta were taken out to make slices with paraffin embedding. Four sections from aortic root of each mouse were chosen to measure and calculate the percentage of lipid core (LC) in the total area of plaque (TP) and the lipid/collagen ratio (L/C) in the plaque by HE and Movat staining respectively, and the mean value of the four sections was taken for analysis. The expressions of granulocyte-macrophage colony-stimulating factor (GM CSF) and tumor necrosis factor-alpha (TNF-alpha) in mice's aorta root were determined by immunohistochemical staining as well. RESULTS: After being treated for 13 weeks, either the percentage of LC in TP and the L/C ratio was significantly lower in all the test drug treated groups than those in the model group, respectively (P < 0.01), especially prominent in the group treated with drug 3. Although lowering of the two indexes presented in all the 3 groups treated by drug 1, 2 and 3, significant difference still presented between drug 3 treated group vs drug 1 and 2 treated group (P < 0.05). As for the expressions of GM-CSF and TNF-alpha, in comparing with the untreated model group, significant decreasing of the TNF-alpha showed only in the drug 4 treated group, while that of GM-CSF could be found in all the test drug treated groups (P < 0.05). CONCLUSION: All the 4 drugs tested in the recommended dosage can stabilize the vulnerable plaques in ApoE knockout mice by improving the constitution of plaque, among them, drug 3 and 4, the drugs possess both the actions of activating blood circulation and detoxicating, show more significant effect, and their mechanisms may be related to their actions in regulating lipid metabolism and inhibiting inflammatory reaction. PMID- 18386575 TI - [Establishment of a quantitative mathematical model of Shen-deficiency syndrome based on whole-genome transcriptional profiles]. AB - OBJECTIVE: To establish a quantitative mathematical model of Shen-deficiency syndrome of TCM by utilizing whole-genome transcriptional profiles. METHODS: The 4, 10, 18, 24 months old SD rats were used, 24-months aged rats intervened by Epimedium Flavonoids (EF) were adopted in the experiment. Rats' hypothalamus, pituitary, adrenal, lymphocytes, bone, liver, and kidney, and spleen were taken for determining whole-genome mRNA expression with gene chip, and a quantitative nerve network model was established by utilizing the gene expression profile of different aged rats, then the model was used to evaluate the effects of EF on Shen-deficiency syndrome. RESULTS: Totally 199 genes showing age-dependent characteristics were screened out from the 7 kinds of tissue, most of them were neuro-endocrine immune related genes. Evaluation based on the mathematical model showed the age of hypothalamus, pituitary, adrenal, liver, kidney, bone, and spleen in the 24-months rats after EF intervention was 12.64, 10.87, 8.10, 12.70, 11.93, 13.14, and 10.13 months, respectively. CONCLUSION: A quantitative mathematical model can be established based on the gene expression profile, it is suitable for estimating the efficacy of Shen-tonifying drugs. EF can make the gene expression of elder close to the young state, suggesting that EF has action in improving Shen-deficiency syndrome and delaying senescence. PMID- 18386576 TI - [Effects of Shenqi Fuzheng injection on gene expression profile of liver tissue with metastatic carcinoma in mice]. AB - OBJECTIVE: To explore the effect of Shenqi Fuzheng Injection (SFI) on gene expression profile of liver tissue with metastatic carcinoma in mice. METHODS: Twenty liver metastatic carcinoma model mice were established by splenectomy after their spleens were injected with 0.2 mL colon cancer C26 cell strain oncocyte liquid, then they were randomly divided into the model group and the SFI group. Starting from the 5th day after modeling, mice in the model group and the SFI group were given via intraperitoneal injection once every other day with physiological saline and SFI respectively. All the mice were sacrificed at the 15th day and the gene profile of metastatic liver carcinoma tissue in the two groups were screened by whole genome chip technique. RESULTS: (1) The model establishing successful rate reached 100%; (2) Gene expression showed that as compared with the model group, in the SFI group, 123 genes were up-regulated, with 52 of them registered to Ensemble, while only one gene was down-regulated and registered to Ensemble was none. CONCLUSION: SFI plays its role of anti-tumor mainly by upregulating several relative genes to promote apoptosis of tumor cells and stabilizing chromosomes. PMID- 18386577 TI - [Effect of Neiyi Kangfu suppository on expressions of cytochrome C and survivin in ectopic and eutopic endometrium in rats with endometriosis]. AB - OBJECTIVE: To explore the different effects of rectal application of Neiyi Kangfu Suppository (NYKFS) on the expressions of cytochrome C (Cyt C) and Survivin in ectopic and eutopic endometrium in endometriosis (EMT) model rats. METHODS: EMT rats were randomly divided into 6 groups, the high- and low-dose NYKFS groups, the Danazol Capsule (DN) group, the Dan'e Fukang Soft Extract (DFE) group, the model group and the blank group. The expressions of Cyt C and Survivin were measured using immunohistochemical SP staining method. RESULTS: (1) After treatment, the IOD value of Cyt C in ectopic endometrium significantly increased in the high-dose and low-dose NYKFS groups respectively to 6.08 +/- 0.35 and 6.23 +/- 0.35, which was significantly higher than that in the model group (5.07 +/- 0.70) and DFE group (5.98 +/- 1.02) respectively (P < 0.05, P < 0.01); while those of Survivin in ectopic endometrium was significantly decreased to 5.73 +/- 0.93 and 5.62 +/- 0.93 and was significantly lower than that in the model group (6.01 +/- 1.16, P < 0.05 or P < 0.01). (2) The lowered eutopic endometrial level of Cyt C in EMT rats after treatment was significantly higher in the two NYKFS groups than that in the model group (P < 0.05); while that of eutopic endometrial Survivin was insignificantly different between the NYKFS groups and the model group or the blank group (P > 0.05). CONCLUSION: NYKFS plays its role in inducing apoptosis by increasing the expression of Cyt C and decreasing the expression of Survivin in ectopic endometrium, but it up-regulates expression of Cyt C, has no obvious effect on Survivin expression in eutopic endometrium. PMID- 18386578 TI - [Effects of erigeron injection on renal interstitial fibrosis in rats]. AB - OBJECTIVE: To investigate the effect and mechanism of Erigeron Injection (EI) on renal interstitial fibrosis in rats. METHODS: Unilateral ureteral obstruction (UUO) model rats were taken as the subject of study. Thirty-six Sprague-Dawley rats were randomly divided into the control group (A), the UUO model group (B) and the treatment group (C) treated with intraperitoneal injection of EI 5 mL/kg per day from 24 h before to 9 days after the operation. On the 10th day of UUO, rats were killed and their kidneys were processed to paraffin sections with HE, PAS and picro-sirius-red staining. The pathological change of renal tubular interstitial tissue and relative cortical/interstitial volume (C/I) as well as the relative content of collagen (RC) were observed by light microscope. The expression of transforming growth factor beta1 (TGF-beta1), alpha-smooth muscle actin (alpha-SMA) and collagen I in the renal mesenchyma were examined by immunohistochemistry. RESULTS: Marked renal interstitial fibrosis changes were found in Group B and C, but the changes were milder in Group C. C/I and RC were higher in Groups B and C as compared with those in Group A (P < 0.01), but they were much lower in Group C than in Group B (P < 0.01). The expression of TGF beta1, alpha-SMA and collagen I were higher in Group B and C than those in Group A (P < 0.05), but they were lower in Group C than in Group B (P < 0.05). CONCLUSION: EI could ameliorate renal interstitial fibrosis in rats, which might be partially realized by down-regulating the expression of TGF-beta1 to prevent the renal epithelial cell differentiation and reducing the synthesis of collagen I. PMID- 18386579 TI - [Study on homologous bone marrow mesenchymal stem cells in repairing peri-tubular capillary cluster]. AB - OBJECTIVE: To investigate the potential effect of homologous bone marrow mesenchymal stem cells (MSCs) on repairing peri-tubular capillary cluster (PTCC), and on improving renal tubular and mesenchymal hypoxia condition. METHODS: Monocyte was purified from bone marrow, amplified and identified as MSCs in vitro. Thirty female Wistar rats were randomly divided into 3 groups, the normal control group (Group A), MSCs transplanted group (Group B) and un-transplanted group (Group C). Rats in Group A was administered with drinking water by gastrogavage for 12 weeks, while those in Group B and C were administered with Aristolochia Decoction for 12 weeks to establish chronic aristolochic acid nephropathy (CAAN) model. At the end of the 12th week, 1 ml of MSCs was injected through caudal vein to the rats in Group B, while to those in Group A and C normal saline was injected instead. Blood, urine and kidney tissue of rats were collected at the end of the 16th week for examination, and their kidney tissue were made into serial section for determining the distribution of Y chromosome and CD34 double positive cells, and the pathological, immunohistochemical changes were observed using Western blotting and RT-PCR, etc. RESULTS: Y chromosome and CD34 double positive cells could be seen in MSCs transplanted renal tissue in group B. At the end of the 16th week, the PTCC density in Group C and B was (26.47 +/- 1.56)/ 0.13 mm2 and (5.26 +/- 0.78)/0.13 mm2 respectively, and the IOD value of hypoxia inducible factor-1alpha (HIF-1alpha) in them was (6.74 +/- 0.67) x 10(3) and (25 27 +/- 1.46) x 10(3) respectively, all showing significant difference between the two groups (P < 0.01). The content of CD34 was higher in Group B than that in Group C (P < 0.01). CONCLUSION: Homologous MSCs can enhance the vascular endothelial cells differentiation to repair the PTCC, thus to improve the renal tubular and mesenchymal hypoxia status. PMID- 18386580 TI - [Effect of Shenqi Fuzheng injection combined with chemotherapy on thirty patients with advanced breast cancer]. AB - OBJECTIVE: To observe the clinical efficacy of Shenqi Fuzheng Injection (SFI) combined with chemotherapy in treating patients with advanced breast cancer. METHODS: Sixty patients were randomly assigned to two groups by digital table, the control group and the treatment group, 30 in each group. All patients were treated with the same CTF regimen of chemotherapy for 21 days as one therapeutic cycle, while those in the treatment group were given SFI additionally in the meanwhite. The therapeutic efficacy was evaluated after 2 cycles of treatment by observing the changes of short-term efficacy, TCM syndrome, quality of life and immune function, as well as the adverse reaction. RESULTS: The total short-term remission rate, the improvement rate of clinical syndrome and quality of life was 50.0%, 70.0% and 76.7% in the treatment group, and 43.3%, 46.7% and 50.0% in the control group, respectively, showing significant difference between the two groups (all P < 0.05). The occurrence of adverse reaction in the treatment group was lower than that in the control group (P < 0.05). The level of CD3+ CD4+ and CD4+/CD8+ ratio increased (P < 0.05) and CD8+ decreased in the treatment group (P < 0.01), while they showed insignificant change in the control group. CONCLUSION: For treatment of advanced breast cancer, SFI can alleviate the bone marrow inhibition caused by chemotherapy, improve clinical symptoms and quality of life and prolong the survival period by regulating cellular immune function of patients, so as to enhance the therapeutic effect of chemotherapy. PMID- 18386582 TI - [Qualitative research and evidence-based medicine]. AB - Qualitative research originates from anthropology and social medicine, it has been introduced into medical field only for ten odd years. As medicine owns social humanity characteristics, its research method should not be limited to the traditional quantitative research for cause-effect relationship, and the advantages of qualitative research is just that it focuses on those unexplained and unanswerable questions by quantitative research. This essay introduces the basic concept of qualitative research, its relationship with evidence-based medicine and applicable scope, and also the necessity and importance of its application in traditional Chinese medicine. PMID- 18386581 TI - [Effect of TCM therapy for removing toxic substance and unblocking meridians on post-radiation quality of life in 55 patients with lung cancer]. AB - OBJECTIVE: To observe the effect of TCM therapy for removing toxic substance and unblocking meridians on post-radiation quality of life (QOF) in 55 patients with lung cancer. METHODS: All patients were randomly assigned to two groups, the 55 patients in the treated group were treated with radiotherapy combined with the TCM therapy, and the 53 in the control group were given radiotherapy alone. The clinical efficacy was evaluated by indexes including KPS scores, QLQ-C30 questionnaire, LC13 specific scale for lung cancer, evaluation criteria for TCM efficacy, the 6-min walking distance, and body weight, etc. RESULTS: The KPS score in the treated group after treatment was improved in 13 patients (23.6%), stabilized in 27 (49.6%) and declined in 15 (27.3%), while in the control group, it was improved in 7 (13.2%), stabilized in 21 (39.6%) and declined in 25 (47.2%), the difference between the two groups was significant (P < 0.05). The results from evaluation with QLQ-C30 questionnaire and LC30 scale in the treated group was also superior to those in the control group (P < 0.05), respectively. The scores of TCM efficacy (24.52 +/- 8.98 vs 29.76 +/- 7.09, P < 0.05) and the 6 min walking distance (233.4 +/- 29.2 m vs 205.8 +/- 31.9 m, P < 0.05) were better in the treated group. After treatment in the treated group, the body weight was increased in 11 patients (20.0%), stabilized in 28 (50.9%) and decreased in 16 (29.1%), while in the control group it was increased in 5 (9.4%), stabilized in 26 (49.1%) and decreased in 22 (41.5%), difference between the two groups was significant, too (P < 0.05). CONCLUSION: TCM therapy for removing toxic substance and unblocking meridians can improve the post-radiation QOL of patients with lung cancer to some extent. PMID- 18386583 TI - [Essential characteristics of qualitative research and its commonly used methods]. AB - The main objectives of qualitative research lies in exploring the opinion, attitude, behavior, and experience of a person as a social role, also a patient. This essay introduces the basic characteristics of qualitative research, including its natural property, inductive method adopted, open character and wholism concept; the results of qualitative research are presented in a text form; and its commonly used methods include observation, individual interview and focus group discussion. PMID- 18386584 TI - [Qualitative research and clinical study on cancer]. AB - Tumor is one of the fateful diseases that human must confront. Currently, quantitative research is still the principal body of the research on cancer. Qualitative research can compensate the limitations of quantitative research in evaluating therapeutic effects on cancer, it can profoundly understand the attitude, experience, confidence, presumable problems and obstacles of doctors and patients to the therapy. This article introduces the necessity and general situation of development in applying qualitative research methods in the researches on tumor, and the main problems and developing tendency for extending in China. PMID- 18386585 TI - [Application of qualitative research method in clinical study on acupuncture and moxibustion]. AB - Clinical qualitative research on acupuncture has turned into one of the overseas attentive points in medical research, and up to now nearly one hundred papers have been published, covering various pivotal aspects of clinical acupuncture itself, such as therapeutic effect evaluation, experience and expectation of patients as well as understanding and experience of acupuncturists on the treatment. Currently, the qualitative research on the holistic therapeutic mode and efficacy evaluation of acupuncture has become the focus of research in foreign countries. Compared with the condition abroad, there is almost no relavent paper published in China. In this paper, the development status, focal field and main direction of overseas qualitative research on acupuncture are introduced, the characteristics of the research proposed in China is analyzed, and its future application in China is looked forward to. PMID- 18386586 TI - [Diagnosis and treatment of traditional Chinese medicine and laboratory medicine]. AB - Traditional Chinese medicine (TCM) and Western medicine have the common purpose and can complement each other, though they belong to different medical systems. Laboratory medicine can provide the scientific bases for modernization and standardization of TCM by offering elaborate and objective laboratory data. Fully playing the role of laboratory medicine in TCM diagnosis and treatment will not only be beneficial to the rush of TCM out of China toward the world, and also be favorable to the innovating and developing of laboratory medicine in integrative medicine. PMID- 18386587 TI - [Progress of study on treatment of Alzheimer's disease with active ingredients of Chinese herbal medicines]. AB - In order to advance the treatment of Alzheimer's disease with active ingredients of Chinese herbal medicines and the research on these ingredients and their effective targets in treating the disease, the relative representative literatures published in recent years were reviewed and summarized in this paper. PMID- 18386588 TI - [Progress in postoperative management of colorectal cancer with integrative medicine]. AB - The comprehensive treatment of colorectal cancer has been quickly progressing in recent years, and TCM treatment plays an active role in the postoperative management of colorectal cancer. In this paper, the progress of postoperative management of colorectal cancer with integrative medicine was reviewed to provide a reference for guiding the researches on this topic. PMID- 18386589 TI - Spinal manipulative therapy in the treatment of low back pain. PMID- 18386590 TI - The TransforMED project. PMID- 18386591 TI - Back on the mat. PMID- 18386592 TI - Pharmacologic management of adult depression. AB - Major depression is a common and treatable disease. Many patients benefit from pharmacologic treatment and, because there is little variation in antidepressant effectiveness, medication choices should be made based on patient characteristics, safety, and anticipated side effects. Most patients respond favorably to treatment, but many do not have complete symptom relief. Changing medications or augmenting with a second medication is helpful for some partial or nonresponders. All antidepressants are capable of producing harmful side effects, and some are particularly prone to dangerous drug-drug interactions. The risk of suicide is always a concern in depression and this risk is not necessarily reduced by the use of antidepressants. Some persons may have an increase in suicidal thoughts with antidepressant treatment. Close follow-up is required when initiating therapy and adjusting dosages. PMID- 18386593 TI - Information from your family doctor. Treating depression: what you should know. PMID- 18386594 TI - Common dental infections in the primary care setting. AB - Family physicians commonly encounter patients with dental infections, such as dental caries and periodontal disease. Dental caries is caused by bacteria that destroy the enamel and dentin; it can be detected by an oral examination that shows stained pits or fissures on the tooth surface. Use of fluoride is the most effective prevention measure for dental caries. Untreated caries may progress to pulpitis and, eventually, to necrosis of the pulp. In irreversible pulpitis, the tooth dies and the patient may have a localized abscess that can spread to surrounding tissue. Periodontal infections are caused by bacteria in the subgingival dental plaque. In gingivitis, the inflamed gums bleed easily with brushing or flossing; the condition can be controlled with good oral hygiene. Periodontitis is characterized by a loss of supportive bone structure caused by chronic gingivitis; it is also associated with some systemic diseases. Localized periodontitis is treated with mechanical debridement and good oral hygiene, whereas generalized periodontitis requires adjunct antibiotic therapy. Pericoronitis results when food particles become trapped under the gum of an impacted tooth. This condition can be controlled by removal of food debris and good oral hygiene. For patients in whom dental infections are disseminated and have invaded the deeper oral spaces, antibiotic treatment should be initiated at the time of referral. PMID- 18386595 TI - Information from your family doctor. Common dental infections. PMID- 18386596 TI - The role of the family physician in the referral and management of hospice patients. AB - Hospice is available for any patient who is terminally ill and chooses a palliative care approach. Because of the close relationship that primary care physicians often have with their patients, they are in a unique position to provide end-of-life care, which includes recognizing the need for and recommending hospice care when appropriate. The hospice benefit covers all expenses related to the terminal illness, including medication, nursing care, and equipment. Hospice should be considered when a patient has New York Heart Association class IV heart failure, severe dementia, activity-limiting lung disease, or metastatic cancer. Timely referrals are beneficial to both patient and hospice because of the cost related to initiating services and the time required to form a therapeutic relationship. Once the decision to refer to hospice is made, the family physician typically continues to be the patient's primary attending physician. The attending physician is expected to remain in charge of the patient's care, write orders, see the patient for office visits, and complete and sign the death certificate. Hospice, in turn, is a valuable physician resource when it comes to medication dosages, symptom management, and communication with patients and their families. PMID- 18386597 TI - Information from your family doctor. Hospice: what you should know. PMID- 18386598 TI - USPSTF recommendations for STI screening. AB - Since 2000, the U.S. Preventive Services Task Force (USPSTF) has issued eight clinical recommendation statements on screening for sexually transmitted infections. This article, written on behalf of the USPSTF, is an overview of these recommendations. The USPSTF recommends that women at increased risk of infection be screened for chlamydia, gonorrhea, human immunodeficiency virus, and syphilis. Men at increased risk should be screened for human immunodeficiency virus and syphilis. All pregnant women should be screened for hepatitis B, human immunodeficiency virus, and syphilis; pregnant women at increased risk also should be screened for chlamydia and gonorrhea. Nonpregnant women and men not at increased risk do not require routine screening for sexually transmitted infections. Engaging in high-risk sexual behavior places persons at increased risk of sexually transmitted infections. The USPSTF recommends that all sexually active women younger than 25 years be considered at increased risk of chlamydia and gonorrhea. Because not all communities present equal risk of sexually transmitted infections, the USPSTF encourages physicians to consider expanding or limiting the routine sexually transmitted infection screening they provide based on the community and populations they serve. PMID- 18386599 TI - Diagnosis of appendicitis: part 1. History and physical examination. PMID- 18386600 TI - Recent-onset muscle hypertrophy unrelated to exercise. Amyloidosis. PMID- 18386601 TI - Therapeutical perspectives for apoptosis modulation in cardiovascular disease. AB - Apoptosis, or programmed cell death, is an active form of cell death, distinct from necrosis, through which multicellular organisms dispose of cells efficiently. Implication of apoptosis in the initiation and progression of many cardiovascular diseases, such as heart failure, systemic hypertension, coronary artery disease, has raised the possibility to elaborate new classes of medication to modulate this process. We review the most important drugs used in cardiovascular diseases and their interference with apoptosis, demonstrated by clinical studies as well as the involved mechanisms. We also analyze new molecules which protect cells from apoptosis and may therefore be clinically useful, as a new class of medication. PMID- 18386602 TI - Current opinions regarding the spinal cord ischemia syndrome. AB - Spinal cord ischemia syndrome (SCIS) is a serious complication which may occur after either internal or surgical diseases in newborn, young children, teenagers and adults; it is also followed by paraplegia. The onset is acute in 95% of cases. In the other cases the onset may be subacute, developing within one week, or it may be chronic, with slow, progressive development, within a few months to a year. The etiology and pathogenesis of this syndrome raises the interest of many medical fields, such as anatomy, physiology, internal medicine, surgical and imaging specialties. In current medical practice the role of spinal arteriography in diagnosing spinal cord ischemia is essential. Arteriography reveals obstructive lesions in the emerging area of the lumber artery, located between T8 and T12 in 85% of patients. Usually, after diagnosing this syndrome, it may be very difficult to reveal the underlying disease and it may require several investigations such as normal and 3D CT scans, SCIS, cerebral or myelic densitometries. This condition may be caused by metabolic congenital or acquired diseases, infectious vascular diseases, osteoporosis: it may also occur after general or peridural anaesthesia or surgical procedures such as spine surgery, neuro- and cardio-vascular surgery, vertebral and myelic trauma and so on. Treatment for this syndrome will be conducted with respect to the underlying disease. Prognosis may depend on patient's age and it is usually difficult to estimate due to the impossibility of determining the type and extent of the medullary lesion (axonotmesis, neurotmesis or other lesions). PMID- 18386603 TI - Quality of life in functional gastrointestinal disorders: assessment and management. AB - Functional gastrointestinal disorders (FGIDs) represent a major burden for the patients, GPs and gastroenterologists. The chronic complaints and psychological disorders associated to the digestive symptoms lead to an impairment of the quality of life. A correct assessment of each functional gastrointestinal disorder is necessary in order to better manage these embarrassing conditions. The advent of Rome III will improve this approach. PMID- 18386604 TI - Psychological perspectives on the pathogenesis of periodontal disease. AB - When defining the relationship between psychosocial factors and periodontitis several issues should be considered: most published studies support a positive association periodontitis, and several psycho-social factors: social isolation, socio-economic status, personality factors, anxiety, depression and life stress; many studies are cross-sectional in nature, include relatively small sample sizes and have inadequate control of potential confounding factors; varying methods to assess both psychosocial factors and periodontitis were used; psychological variables are usually measured by self-report scales and the informers may supply incorrect information and the situation bias may also take place that is the condition of instability of the clinical phenomenon being evaluated; the demographic makeup of the population under study (age, gender, education level, race) and control of potential confounding variables (smoking, oral hygiene status) differs markedly across studies. The findings indicate that the main mechanisms by which psychosocial factors promote gingivitis and periodontitis development included: 1) behavioural mechanisms, whereby the given factor exacerbates lifestyles known to potentiate periodontal disease (e.g., neglect of oral hygiene, changes in diet, increase in smoking), and 2) direct pathophysiological effects on host resistance. As a result, interferences on the association between psychosocial factors and periodontal disease require careful considerations. PMID- 18386605 TI - Is acute myocardial infarction a localized or rather a global coronary phenomenon? AB - BACKGROUND: Use of qualitative assessment of coronary artery flow (TIMI), although widely spread, represents a subjective method, a quantitative assessment (CTFC) being necessary in order to standardize and facilitate comparisons and communications of angiographic trials. This study aims at appreciating whether myocardial infarction represents a global phenomenon that affects the whole myocardium, also affecting the coronary artery flow in non-culprit arteries. MATERIALS AND METHODS: 66 patients that underwent primary PCI with stent and 66 patients with normal angiographic coronary arteries were studied. The number of frames necessary for the dye to reach certain standardized landmarks was registered, in order to objectively assess the coronary artery flow as a continuous variable. The statistical evaluations revealed that the quantitative assessment of the coronary flow through CTFC (corrected TIMI frame count) pins up a difference between the flow on non-culprit coronary arteries (23.54+/-9.235) and the flow on normal angiographic coronary arteries (17.46+/-4.1) (p<0.005), the result being also valid for each of the three coronary arteries separately analyzed: LAD (23.88+/-8.08 vs. 18.575+/-4.59 - p<0.005), CX (20.7+/-7.34 vs. 15.62+/-3.35 - p<0.005) and RCA (26.45+/-11.91 vs. 18.2+/-3.69 - p<0.005). CONCLUSIONS: There are significant differences regarding the flow on non-culprit coronary arteries in patients that suffered acute myocardial infarction (AMI) and the flow on the coronary arteries of the patients with normal angiographic results; these data might be the expression of global myocardial suffering. PMID- 18386606 TI - The effect of trimebutine on the psychosocial adjustment to illness in the irritable bowel syndrome. AB - BACKGROUND AND AIM: Irritable bowel syndrome (IBS) is a harassing condition leading to the impairment of the quality of life. Adjustment to illness is an important personality factor able to influence the outcome. We looked for the psychological adjustment to illness in IBS. We also investigated the effect of trimebutine, a drug frequently used in IBS, on symptoms and psychosocial adjustment to illness in IBS. METHODS: A prospective randomized controlled trial including 50 consecutive patients with irritable bowel syndrome. They received either trimebutine 100 mg t.i.d (n=25, 11M/14F, aged 44+/-12 years) or placebo t.i.d. (n=25, 12M/13F, 48+/-10 years) for 2 weeks. Seven digestive symptoms were evaluated on a 0-16 scale, as well as the patient's satisfaction (scale 1-10) and the physician's satisfaction (scale 0-4) with the therapy. The psychosocial adjustment to illness was assessed by the PAIS-SR scale (self-report). RESULTS: Trimebutine increased significantly the following items of the PAIS-SR questionnaire: vocational environment, domestic environment, sexual relationships, extended family relationships, social environment but not health care orientation and psychological distress. Symptom score (mean+/-SD) for pain was reduced from 13.1+/-4.0 to 2.7+/-1.1, p<0.0001 by trimebutine and from 12.5+/ 3.6 to 7.7+/-2.5, p<0.05 by placebo. Bloating score was decreased by trimebutine from 10.3+/-3.6 to 2.6+/-1.2 (p<0.0001) and not by placebo (from 10.5+/-4.5 to 8.8+/-3.6 N.S.). Nausea, emesis, anorexia, diarrhea and constipation were also improved by trimebutine. Placebo improved anorexia and tended to improve nausea. Patient' s satisfaction score was 8.1+/-1.1 with trimebutine and 5.2+/-3.5 with placebo (p<0.01). Physician's satisfaction score was 3.0+/-0.7 with trimebutine and 2.2+/-1.1 with placebo (p<0.02). CONCLUSIONS: Trimebutine improved symptoms and psychological adjustment to illness in irritable bowel syndrome in comparison with placebo. PMID- 18386607 TI - Renovascular hypertension in children: symptoms and manifestations. A 19 patients clinical study. AB - The symptomatology in renovascular hypertension in children may be sometimes misunderstood, leading to the establishment of a late diagnosis, even after the inducing disease is diagnozed. METHODS: The main types of manifestations on which the diagonsis was established: neuropsychiatric, weight loss, renal and digestive manifestations were studied in 19 patients, age ranged 2 to 15 years old in which the main causes diagnosed were reno-ureteral malformations, pyelonephritis, reno ureteral reflux and renal trauma. Starting from these observations the diagnosis of arterial hypertension was established and the further investigations have shown the real causes of arterial hypertension noticed. RESULTS: The periodic postoperative evaluation performed every three months over a peri6d ranging from 4 months to 7 years individualized 4 clinical evolutive types: Arterial Hypertension with lumbar pain, without any clinical manifestation, with ophthalmologic manifestations and with encephalopathic manifestations. CONCLUSIONS: Symptomatology in renovascular hypertension has no pathognomonic indices and the observed manifestations being single or all together impose the evaluation of blood pressure or even more, the monitoring of it over a long period, for at least 30 days. If the values of blood pressure are greater than the normal accepted ones, more complex investigations are required in order to establish a certain etiology of the hypertensive status. PMID- 18386608 TI - Hypertension in lupus nephritis. AB - Systemic lupus erythematosus, an inflammatory disease, has lupus nephritis as severe complication. Hypertension, secondary of the lupus nephritis represents a major cardiovascular risk factor. The impact of hypertension on the evolution and prognosis in lupus nephritis patients as well as the need to initiate pathogenic, antihypertensive and cardiovascular risk factors therapy have been evaluated. PMID- 18386609 TI - Evaluation of granulocyte elastase as a sensitive diagnostic parameter of inflammation in first ischemic stroke. AB - Several traditional vascular risk factors are associated with proinflammatory alterations, including leukocyte activation, and predispose cerebral vasculature to thrombogenesis on inflammatory stimulation. PMN elastase derived from the activated neutrophils might play an important role in injury. This study tries to elucidate the involvement of activated polymorphonuclear neutrophil proteolytic activity in the pathophysiology of ischemic stroke. Levels of PMN elastase/alpha 1-proteinase inhibitor complex (PMN-E/alpha1-PI) were measured in 145 patients (mean age 72.4+/-6.3) with first ischemic stroke from the onset day (day 0) to 7 days after onset (day 7) and 260 control subjects drawn matched for age, sex. Quantitative estimation of the plasma levels of the PMN-E/alpha1-PI complex was carried out with a highly sensitive enzyme-linked immunoassay (ELISA). Plasma PMN elastase levels were statistically significantly higher on 7 day patients (384.7+/-31.4 microg/L vs 216.5+/-26.8 microg/L on day 0; p<0.01) than in those without ischemic stroke (67.4+/-5.2 microg/L). Plasma PMN elastase may be a well characterised and sensitive inflammation marker, and it is associated with endothelial dysfunction. PMN elastase can be used as a measure for the activity of granulocytes during an inflammatory response. Furthermore it may be possible to treat such disorders with proteinase inhibitors. PMID- 18386610 TI - The discovery by Gh. Benga of the first water channel protein in 1985 in Cluj Napoca, Romania, A few years before P. Agre (2003 Nobel Prize in Chemistry). AB - The first water channel protein, now called aquaporin 1, was identified or "seen" in situ in the human red blood cell membrane by Benga's group in 1985. It was again "seen" when it was by chance purified by Agre'group in 1988 and was again identified when its main feature, the water transport property, was found by Agre's group in 1992. Consequently, the omission of Gh. Benga from the 2003 Nobel Prize in Chemistry (half of which was awarded to P. Agre "for the discovery of the water channels") is a new mistake in the award of Nobel Prizes. The growing recognition of the priority of Gh. Benga over P. Agre in the discovery of water channels is documented in this paper. PMID- 18386611 TI - The analgesic effect of gemcitabine in mice. AB - The analgesic effect of gemcitabine was analyzed in two experimental tests in mice. In the first test gemcitabine produced a dose-dependent decrease of the number of torsions produced by i.p. acetic acid in mice proving an analgesic effect. In the hot plate test the gemcitabine produced a non-statistical significant increase of the latency of the leakage, but a statistically significant increase of the latency of the salt proving an analgesic effect produced especially by a nervous central mechanism like morphine. The association of the gemcitabine and morphine proved that an inactive dose of gemcitabine increased statistically significant the effect of morphine proving a potentiation of the effect of morphine by gemcitabine. A same potentiation was demonstrated also by an effective analgesic dose of gemcitabine. The analgesic effect of the gemcitabine was antagonized by aminophiline, an inhibitor of the phosphodiesteraze, proving that the analgesic effect of the gemcitabine could be produced by a decrease of the intracellular cAMP. According to the chemical structure of gemcitabine it is possible that gemcitabine decrease the velocity of the degradation of the GMP fixed on the alpha subunit of the Gi protein and so increase the effect of the activated a subunit of the Gi protein to inactivate the adenylatecyclase. The analgesic effect of gemcitabine could be important in clinical management of antineoplastic patient by increasing the quality of life of these patients and the property of gemcitabine to potentiate the analgesic effect of morphine could be important by decreasing the needs of morphine of the antineoplastic patients. More researches will be necessary for evaluating also other nervous central effects of gemcitabine, for evaluation if other antineoplastic drugs have the same effect and for establishing a possible chemical structure-analgesic effect relationship and maybe also for evaluation if other effects of morphine are potentiated by gemcitabine and other antineoplastic drugs. PMID- 18386612 TI - Effect of dietary isoflavones on plasma cholesterol. AB - The effects of dietary isoflavones on plasma cholesterol are discussed in order to find new diet therapies for hypercholesterolemia, which represents a cardiovascular risk factor. Isoflavones represent a class of biological substances with beneficial action in the reduction of cholesterol, LDL cholesterol, increase of HDL cholesterol and reduction of triglycerides as well as other risk factors of atherosclerotic cardiovascular diseases. The cholesterol reducing effects of isoflavones have been demonstrated by numerous studies that started from the finding that the Japanese, consumers of more proteins of vegetal origin, present a lower incidence of ischemic heart disease, the mortality rate by cardiovascular disease being about half of that of the U.S.A. Subsequent studies were carried out in patient groups with high, average, small, or normal cholesterol levels, adults or children, all of them reporting that a diet containing vegetal isoflavones led to the reduction of cholesterol levels. PMID- 18386613 TI - Myocardial infarction in young patients. AB - Myocardial infarction (MI) is relatively rare in young patients. Atherosclerosis is responsible for most cases, but in one fifth of reported events other causes of MI are involved. Regarding individual susceptibility, it seems that cigarette smoking is the most common modifiable risk factor; family history and lipid abnormalities can also play an important role. In the absence of obvious risk factors a careful search for other contributing factors (such as vasospasm, vasculitis, cardiac masses or hypercoagulable states) is warranted. PMID- 18386614 TI - Von Recklinghausen's disease--elements of clinical practice. AB - Neurofibromatosis is a condition described for the first time by Von Recklingahausen in 1882. This disease is one of the most common genetic disorders and is characterized by multiple tumors of the central and peripheral nervous system, brown macules on the skin, bone deformities, and by vascular and visceral disorders. Despite extensive basic science research, the diagnosis is still based largely on clinical criteria, which often present gradually during childhood. PMID- 18386615 TI - Romanian subjects entered in the hypertension in the very elderly trial (HYVET). AB - The baseline characteristics, possibly related to cardiovascular outcome, in 94 Romanian hypertensive men and 168 women over the age of 80 entered into the Hypertension in the Very Elderly Trial (HYVET) were compared with those in 725 men and 1128 women entered into the trial in the rest of Western and Eastern Europe (mainly Russia and Bulgaria). There was a tendency for more women in the Romanian subjects (66.7% versus 60.9% (P = 0.08)) and therefore all comparisons were adjusted for this non significant difference. The Romanian subjects were slightly younger, had more previous hypertension, but more previous strokes than the rest of Europe, and consumed a similar amount of alcohol. The Romanian subjects were, on average, lighter and shorter. Romanians had higher mean concentrations of blood urea and glucose and higher concentrations of HDL cholesterol. Total cholesterol was, on average, lower in Romanian subjects than was serum creatinine. Serum sodium, blood hematocrit and hemoglobin were all, on average, lower in Romanian subjects. The Romanian subjects were recruited from predominantly rural areas and it is hypothesised that their diet may have contained less salt, iron and other haematinics. Calorie restriction, compared to the rest of Europe, may have resulted in lower stature and weight but recent increases in calorie intake may have resulted in the increasing average blood glucose in both sexes. DDT is known to block the uptake of HDL cholesterol by the liver and may explain the increased HDL concentrations in Romania. Some of these biochemical differences may reflect different methodologies employed in Romanian laboratories, but we could not confirm this possibility. PMID- 18386616 TI - Arterial stenosis and atherothrombotic events in polycythemia vera and essential thrombocythemia. AB - Patients with polycythemia vera (PV) and essential thrombocythemia (ET) are at risk of developing arterial and venous thromboembolic complications. Given the complex interaction between blood cells and the vessel wall, it is possible that atherogenesis may also be accelerated in these patients. We used Doppler arterial ultrasound to assess the presence of arterial stenosis in a cohort of PV and ET patients. MATERIAL AND METHODS: A total of 37 patients, 29 with PV and 8 with ET, were investigated. Aside from an extensive clinical and hematological evaluation, arterial Doppler ultrasonography was performed in all patients; in 3 patients arteriography/coronarography was also performed. RESULTS: Twenty four patients (65%) had a history of atherothrombotic events including cerebral ischemic attacks (CIA) in 12 patients, ischemic heart disease (IHD) in 10 patients and peripheral occlusive arterial disease (POAD) in 12 patients. Eight patients had multiple atherothrombotic events. Twenty five patients (67%) had other atherosclerotic risk factors such as smoking, dyslipidemia, hypertension and diabetes. Significant arterial stenosis was found in 23 patients (62.1%), including 12 patients with carotid plaques, 10 with peripheral arterial stenosis, 3 with coronary stenosis, 2 with aortic plaque and 2 with common iliac artery stenosis. In 12 patients multiple arterial stenoses were found. The presence of arterial stenosis was significantly correlated with the occurrence or thrombotic events (p = 0.0003) and was also correlated with the concomitant presence of polyglobulia and thrombocytosis. Both the thrombotic risk and the probability of stenosis detection were augmented by additional risk factors such as smoking, hypertension and dyslipidemia. DISCUSSION AND CONCLUSIONS: The high incidence of arterial thrombotic events in our PV and ET patients was associated with a high incidence of stenosis detectable by arterial ultrasound. Hyperviscosity, endothelial damage due to leukocyte activation with subsequent thrombus formation, hyperhomocysteinemia and hyperexpression of activating genes such as JAK2 and STAT5 are all features characteristic of PV and ET that may contribute, along with other risk factors, to the development and progression of atherothrombosis. Cytotoxic treatment in PV and TE may be beneficial both through its antiproliferative effect on hematopoiesis and on the atherosclerotic plaques, atherogenesis being described as a proliferative disease of the vessel wall. PMID- 18386617 TI - Frequency and prognostic significance of QT prolongation in chronic renal failure patients. AB - Cardiovascular mortality and morbidity are high in chronic renal failure (CRF) patients. Previous studies on non-uraemic patients with heart failure (HF), hypertension or diabetes mellitus (DM) showed that QT and QTc prolongation and dispersion represent cardiovascular risk factors. The patients with long QT interval have more often ventricular premature beats and sudden death than those with normal QT interval. The aim of our study was to evaluate the frequency and predictive value of QT and QTc prolongation in CRF patients, included or not in a chronic dialysis programme. On 68 patients (M/F = 36/32, mean age = 47.6 years), with CRF we analyzed QT and QTc interval with a digital 12 lead electrocardiogram CARDIAX. 8/68 patients (11.8%) had long QT interval (>0.45"). After having calculated QT corrected (QTc) interval according to the heart rate, 28/68 patients (41.2%) had QT prolongation (>0.45"). Multivariate statistical analysis of clinical factors, but also of biological, electrocardiographic, echocardiographic data and 24 hours of ECG and blood pressure monitoring showed that QT prolongation is statistically significant (p < 0.05) correlated with: number of years of renal failure (p = 0.0001), serum concentrations of potassium and calcium (p = 0.0001) and diastolic BP (p = 0.05). QT prolongation in CRF patients is not dependent on the level of uremia or the type of chronic renal substitution (hemodialysis or continuous ambulatory peritoneal dialysis). There were no statistically significant correlations between QT prolongation and serum concentrations of Mg, PO4, HCO3 and Hb. Long QT interval was not dependent either on the dipper/nondipper profile of mean BP values or ejection fraction of left ventricle. In our study long QT interval was not statistically significant correlated with arrhythmias or sudden death. Despite the high incidence of QT prolongation in CRF patients (41.2%), short-term consequences are not as severe as those in cardiac patients. This is possibly explained by the different pathogenic mechanisms of arrhythmia in CRF when electrolytic disorders are the main cause for the development of arrhythmia. During a mean follow-up of 3.8 months (3-5.5) there were no cases of sudden death on patients with QT prolongation, and arrhythmia incidence was not statistically significant higher than in subjects with normal QT interval. PMID- 18386618 TI - Glycoregulation disorders and alterations of C reactive protein in nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological condition which ranges from simple steatosis and steatohepatitis to cryptogenetic cirrhosis. As insulinresistance plays a central pathogenetic role, NAFLD is regarded as the hepatic feature of the metabolic syndrome. Type 2 diabetes mellitus and obesity are the conditions most frequently associated with NAFLD. Subclinical inflammation has been suggested as a possible connective mechanism between glycoregulation disorders and NAFLD. The purpose of our study was to evaluate the prevalence and severity of glycoregulation disorders (impaired glucose tolerance and type 2 diabetes mellitus) in patients with ultrasonographical certified NAFLD, and to determine medium levels ofC reactive protein in these individuals. MATERIAL AND METHODS: The study was conducted on 104 patients with ultrasonographical certified steatosis, divided into two subgroups: simple steatosis and steatohepatitis, based on the elevation of hepatic enzymes. A control group of 100 subjects without ultrasonographical proof of hepatic steatosis had been used. Fasting glucose, oral glucose tolerance test and C reactive protein levels were performed for each patient. Statistical analysis was based on student t-test and Hi-square test. RESULTS: The prevalence of glycoregulation disorders was significantly higher in patients with NAFLD than in controls (p < 0.05). Patients with steatohepatitis had a significantly higher prevalence of type 2 diabetes mellitus than those with simple steatosis. Medium levels of C reactive protein were significantly higher in patients with NAFLD than in controls, respectively in subjects with steatohepatitis than in those with simple steatosis. CONCLUSIONS: NAFLD is at risk for developing glycoregulation disorders compared with controls; the severity of liver damage is correlated with an augmentation in the severity of glycoregulation disorders. Patients with NAFLD have higher levels of C reactive protein compared with controls, the medium levels C reactive protein increasing with an increase in hepatic damage. PMID- 18386619 TI - Prevalence of anti-neutrophil cytoplasmic antibodies in patients with chronic hepatitis C infection associated mixed cryoglobulinemia. AB - Chronic hepatitis C virus (HCV) has been linked to extrahepatic autoimmune phenomena. In addition, a variety of autoantibodies are found in patients with HCV. Anti-neutrophil cytoplasmic antibodies (ANCA) have been considered to be highly specific for systemic small vessel vasculitides with renal involvement. The prevalence, nature, a clinical significance of anti-neutrophil cytoplasmic antibodies (ANCA) in serum samples of patients with HCV were therefore investigated. A prospective study of 67 consecutive patients with chronic HCV with no evidence of previous hepatitis B virus (HBV) infection or any other autoimmune disorder. Twenty-seven patients with HBV and 35 healthy volunteers matched for age and sex served as control groups. Anti-neutrophil cytoplasmic antibodies in the serum samples were measured by indirect immunofluorescence assay. The group of patients with HCV and ANCA in their serum showed significantly higher total IgG levels, a higher incidence of antinuclear antibodies, and viraemia (HCV RNA) than the ANCA negative patients. None of the patients with HCV and ANCA showed any clinical manifestations related to those autoantibodies. This study clearly shows a high prevalence of ANCA in the serum of patients with HCV. The role of these autoantibodies on the course of HCV infection and their clinical significance has not been determined yet. In conclusion, ANCA are not only markers of small vessel vasculitides, but can also be (false?) positive in patients with HCV infection. A hepatitis C associated vasculitis should be considered in the differential diagnosis. Determination of target antigens for ANCA is indispensable. Interpretation and therapeutic decisions concerning ANCA associated HCV should only be performed within the clinical context. PMID- 18386620 TI - Oxidative stress parameters in hemodialysis patients with or without diabetes. AB - Oxidative stress (imbalance of antioxidant and prooxidants in favour of the later) is considered to be a feature of diabetes and chronic renal failure. Carbonyl stress defined as accumulation of reactive carbonyl compounds due to excess production or disturbed clearance from the body is thought to amplify oxidative stress in these conditions. The accumulation of carbonyl compounds can be also a consequence of oxidative stress. A vicious cycle can thus be formed. We have studied the association between carbonyl stress markers (dicarbonyl compounds, Amadori products) and oxidative stress markers (total plasmatic thiols and malondialdehyde level) in hemodialysed patients with or without diabetes taking into account the levels of possible excess substrates (glucose and triglycerides). We have concluded that hemodialysed diabetes patients are more susceptible to oxidative stress than hemodialysed patients without diabetes. PMID- 18386621 TI - Is IGF-1 involved in the regulatory modifications of cholesterolemia following the administration of embryonary peptides? AB - Significant modifications of IGF-1 and cholesterol (total and LDL) were observed following the administration of an extract of embryonary peptides (EP) to old subjects for 60 days. For most of the subjects, due to the aging process, the initial values of the biochemical parameters were shifted towards pathological range. Following the administration of EP, the serum levels of IGF-1 and cholesterol (total and LDL) were shifted towards the physiological limits for their age. The most significant modifications towards physiological range were observed for subjects with high, initial levels of IGF-1, when the decrease was striking (1-2 orders of magnitude). For these subjects, significant modifications were observed simultaneously for cholesterol. The modifications induced following the administration of EP exhibit a regulatory feature, as they are dependent on the initial levels of these parameters. The action of EP on the levels of IGF-1 and cholesterol was significantly equal for both sexes, but the influence of EP was more clear-cut in men. In conclusion, our results support an implication of IGF-1 in the regulatory mechanisms of cholesterolemia in old subjects following the long-term administration of EP. PMID- 18386622 TI - Fabry's disease. AB - Fabry's disease is a rare X-linked, recessive, glycolipid storage disorder. It is caused by the deficient activity of a lysosomal enzyme, alpha-galactosidase A. Deficiency of alpha-GAL causes an inability to catabolize the lipids with cellular accumulation of its most abundant substrate, globotriaosylceramide (GL 3), and other neutral glycosphingolipids in the vascular endothelium and numerous tissues throughout the body. This progressive glycosphingolipid accumulation leads to life-threatening clinical sequelae in renal, cardiac and cerebrovascular systems. Heterozygous Fabry's disease is less studied. We present a patient, 43 years old, with cardiac (hypertrophic cardiomyopathy), neurological (sensitive motive polyneuropathy), digestive (chronic diarrheea), renal and cutaneous involvements. PMID- 18386623 TI - A rare case of pulmonary emphysema. AB - Alpha-1-antitrypsin deficiency (AAT) is one of the three most common lethal genetic diseases in the caucasian population (together with cystic fibrosis and Down syndrome). Its primary manifestation is early-onset panacinar emphysema. Slowly progressive dyspnea is the primary symptom, although some patients initially have symptoms of cough, sputum production, or wheezing. A minority of patients develops hepatic cirrhosis. We present a case of a 40 year-old male, light smoker, with chronic obstructive lung disease with predominance of panacinar emphysema, with AAT deficiency (72 mg/dl; normal values = 200-300 mg/dl) complicated with cor pulmonale and chronic respiratory failure. The main clinical consequence of AAT deficiency is the early onset of panacinar emphysema, typically more severe at the lung bases. Smoking plays an important part in the natural history of the disease, both increasing the severity and decreasing the age at onset of emphysema. PMID- 18386624 TI - Vibroarthrography--a possible functional non-invasive method for early detection damaged cartilage joint. AB - The vibroacoustic signals emitted by the diarthrodial articulation during a normal movement differentiates a healthy case from a pathologic one, due to alteration in forms and contact surfaces. The differences are shown by a various dynamics of the vibroacoustic spectra. Parametric representation of the acquired signals, filtering and clinical interpretation of those allow classification and fast recognition of a normal/pathologic status of the investigated articulation by the physician. Our method of acquiring the sound and vibration signals is effectuated completely non-invasive, with a set of translators using Pulse - sounds and vibrations analyzers (a matrix of prepolarised microphones with measurement domain in infrasound scale and piezoelectric acceleration transducers). A comparative study of vibroacoustic and thermal spectra, with early alterations revealed by knee nuclear magnetic resonance, correlates vibroacoustic and thermic spectra alterations with morphological ones, by determining certain limits between normal and pathological morphofunctional patterns. This prospective comparative study will help us evaluate the method in terms of sensibility, specificity, negative and positive predictive value, indices that assure the diagnostic power to the method. PMID- 18386625 TI - Discal hernia in children and teenagers: medical, surgical and recovery treatment. AB - Lumbar disc hernia represents a rare situation for the physician. The first intervention in disc hernia was performed during the '40. The rate of surgery needing lumbar hernia is about 1-2%. Lumbar disc hernia in children and teenagers has 4 main causes: familial history, trauma, congenital malformation of the spine and disc degeneration. The symptoms in young patients are dominated by local or ischiadic irradiated pain, but neurological discrepancies rarely occur. PMID- 18386626 TI - [Molecular and genetic analysis of spinocerebellar ataxia type 10 (SCA10)]. AB - Spinocerebellar ataxia type 10 (SCA10) is a dominantly inherited neurodegenerative disease caused by expansion of the ATTCT pentanucleotide repeat in intron 9 of a novel gene, ATXN10, on chromosome 22q13.3. It is clinically characterized by progressive ataxia, seizures, and anticipation, which can vary within and between families. The length of the expanded ATTCT repeats is highly unstable on paternal transmission and shows a variable degree of somatic and germline instabilty, revealing complex SCA10 genetic mechanisms. Moreover, the purity of the expanded repeat element may be a disease modifier. ATTCT repeats have been recently shown to form unpaired DNA structure and may serve as an aberrant DNA replication origin, potentially contributing to repeat instability and cell death. How this untranslated ATTCT expansion leads to neurodegeneration has been still controversial. We discuss several possible pathogenic mechanisms for SCA10, and growing number of evidence indicates a gain-of-function RNA mechanism, similar to the myotonic dystrophies caused by non-coding CTG or CCTG repeat expansions. PMID- 18386627 TI - [Clinical and pathological study on early diagnosis of Parkinson's disease and dementia with Lewy bodies]. AB - [123I] Meta-iodobenzylguanidine (MIBG) myocardial scintigraphy has been used to evaluate postganglionic cardiac sympathetic innervation in heart diseases and some neurological disorders. To see clinical usefulness of MIBG myocardial scintigraphy to differentiate Parkinson's disease (PD) and dementia with Lewy bodies (DLB) from related movement disorders and Alzheimer disease (AD), we performed MIBG myocardial scintigraphy in patients with these disorders. Cardiac uptake of MIBG is specifically reduced in PD and DLB, and this imaging approach is a sensitive diagnostic tool that possibly differentiates PD and DLB from related movement disorders and AD. To see pathological basis of the reduced cardiac uptake of MIBG in Lewy body disease, we immunohistochemically examined cardiac tissues from patients with PD, DLB, related movement disorders and AD using antibodies against tyrosine hydroxylase (TH) and phosphorylated neurofilament (NF). Not only TH- but also NF-immunoreactive (ir) axons in the epicardial nerve fascicles were markedly decreased in Lewy body disease, namely cardiac sympathetic denervation, which accounts for the reduced cardiac uptake of MIBG in Lewy body disease. Patients with PD and DLB have Lewy bodies (LBs) in the nervous system, whereas patients with multiple system atrophy (MSA), progressive supranuclear palsy, corticobasal degeneration, parkin-associated PD and AD have no LBs in the nervous system. Even in patients with MSA, cardiac sympathetic denervation was associated with the presence of LBs. Therefore, cardiac sympathetic denervation is closely related to the presence of LBs in a wide range of neurodegenerative processes. Taken together, we conclude that the reduced cardiac uptake of MIBG is a potential biomarker for the presence of LBs. Because alpha-synuclein is one of the key molecules in the pathogenesis of PD, we further investigate how alpha-synuclein aggregates are involved in degeneration of the cardiac sympathetic nerve in PD. We immunohistochemically examined cardiac tissues from patients with incidental Lewy body disease (ILBD) and PD using antibodies against TH and phosphorylated alpha-synuclein. We found that (1) alpha synuclein aggregates in the epicardial nerve fascicles, namely the distal axons of the cardiac sympathetic nerve, were much more abundant in ILBD with preserved TH-ir axons than in ILBD with decreased TH-ir axons and PD; (2) alpha-synuclein aggregates in the epicardial nerve fascicles were closely related to the disappearance of TH-ir axons; (3) in ILBD with preserved TH-ir axons, alpha synuclein aggregates were consistently more abundant in the epicardial nerve fascicles than in the paravertebral sympathetic ganglia (pSG); and (4) this distal-dominant accumulation of alpha-synuclein aggregates was reversed in ILBD with decreased TH-ir axons and PD, which both showed decreased or depleted TH-ir axons but more abundant alpha-synuclein aggregates in the pSG. These findings indicate that accumulation of alpha-synuclein aggregates in the distal axons of the cardiac sympathetic nervous system precedes that of neuronal somata or neurites in the pSG and that heralds centripetal degeneration of the cardiac sympathetic nerve in PD. This chronological and dynamic relationship between alpha-synuclein aggregates and distal-dominant degeneration of the cardiac sympathetic nervous system may represent the pathological mechanism underlying a common degenerative process in PD. PMID- 18386628 TI - [Case of unilateral alteration due to hypertensive encephalopathy]. AB - We report a patient of a 20-year-old woman of Takayasu's arteritis and hypertensive encephalopathy. The symptoms started with headache and vomiting following status epilepticus. On arrival at the emergency room in our hospital, fever was apparent and cerebrospinal fluid examination revealed pleocytosis. After the admission, the patient presented with hypertension, decreased right brachial pulse and the difference between bilateral brachial arterial blood pressures on examination. There had been no history of arterial hypertension. The MR angiography revealed stenoses of the bilateral cervical, especially right cervical, right middle cerebral and left renal arteries. Brain MRI showed transient hyperintense lesions of the left fronto-parieto-occipital cortices and subcortical white matter in FLAIR and diffusion weighted images. These alterations suggested the presence of reversible vasogenic edema induced by hypertensive encephalopathy. We need to be aware of young patients with convulsion, especially young women, who has arterial hypertension as well as the difference with blood pressures between extremities. PMID- 18386629 TI - [Fulminant strongyloidiasis successfully treated by subcutaneous ivermectin: an autopsy case]. AB - We report a 49-year-old man who was a human T-cell leukemia virus type 1 (HTLV-1) carrier, born in Okinawa prefecture where both strongyloidiasis and HTLV-1 are endemic. He presented with fever, headache and urinary retention. On the basis of CSF examination and MRI findings, his condition was diagnosed as myelitis. He received methylprednisolone pulse therapy. He was transferred to our hospital due to severe paralytic ileus. Strongyloides stercoralis (S. stercoralis) was found in the duodenal stained tissue of a biopsy specimen. Ivermectin applied both orally and through enema were ineffective because of severe ileus and intestinal bleeding. Nine mg (200 microg/kg) of ivermectin solution was administered subcutaneously every other day for five days (total amount 45 mg). The S. stercoralis burden in the stool decreased and paralytic ileus gradually resolved. Three weeks after the resolution of S. stercoralis infection, purulent meningitis developed and acute obstructive hydrocephalus appeared. The hydrocephalus improved by ventricular drainage. Approximately three months after drainage, he died of incidental aspiratory pneumonia. Autopsy showed neither eggs nor larvae of S. stercoralis in the organs. In this case, the fourth reported case in the world, subcutaneous ivermectin injection was dramatically effective. We should consider a diagnosis of strongyloidiasis for any patient from Okinawa prefecture who was an HTLV-1 carrier presenting with unknown origin ileus after treatment of steroid therapy. PMID- 18386630 TI - [Case of eosinophilic myositis in continuum from localized nodular myositis]. AB - We report a 72-year-old man with eosinophilic myositis (EM). At age 71 he noticed a painful nodule in his left calf. A biopsy (first biopsy) showed marked infiltration of mononucleated cells and necrotic muscle fibers. Several phagocytosed fibers were also seen. He was diagnosed as having myositis. The painful nodule disappeared spontaneously. At age 72, he again had a painful nodule, but this time in his right calf; again, this disappeared spontaneously on the first admission. Just after discharge, he noted painful nodules in the left thigh and right anterior tibial muscles and was again admitted (second admission). Neurological examination revealed mild proximal-dominant weakness in all four extremities but no other abnormalities. Laboratory studies showed elevated creatine kinase (CK) level (38,803 U/l; normal 62-287) and positive Jo-1 antibody, but no eosinophilia. Needle electromyography of the limb muscles showed myogenic patterns. Magnetic resonance imaging of the lower limbs demonstrated several T2-high and gadolinium (Gd)-enhanced lesions. Muscle biopsy (second biopsy) from the left quadriceps femoris showed marked infiltration of eosinophils; he was diagnosed as having EM. Administration of prednisolone was initiated at 60 mg/day and then gradually tapered. After starting treatment with steroids, his muscle weakness gradually ameliorated, CK level dramatically decreased, and the nodules disappeared. Clinically, the patient had developed localized nodular myositis (LNM), but pathologically it was EM without peripheral blood eosinophilia and positive Jo-1 antibody that is occasionally found in polymyositis (PM). Thus, this patient demonstrated overlapping characteristics of EM, LNM, and possibly PM, suggesting that a common mechanism underlay these conditions. As discussed, the involvement of eosinophils in three inflammatory myopathies was indicated. PMID- 18386631 TI - [Case of post-stroke dementia after left medial occipitoparietal lesion]. AB - We report herein a patient of poststroke dementia (PSD) following left medial occipitoparietal hemorrhage triggered by drainage of an acute traumatic subdural hematoma. The patient, an independent 73-year-old man, became dependent due to dementia. Cognitive dysfunction was characterized by moderately decreased IQ, severe memory disturbances, topographical disorientation, executive dysfunction and loss of self-awareness. Cognitive dysfunction has not advanced for 3 years. Hypo-perfusion in the whole brain, particularly in the left temporal and parietal regions, was visible on 123I-IMP SPECT images. Magnetic resonance imaging demonstrated damage to the left posterior cingulate cortex, cingulate bundle, superior parietal lobule and subcortical region of the occipital lobe. The fornix was spared. Some subcortical small spotted lesions were detected, but periventricular lucency was not prominent. Structures known to be important in memory but spared by the lesion included the thalamus and basal forebrain. Small spotted subcortical lesions were detected in bilateral hippocampi, which are also known to be important in memory, but these were probably silent lesions. This case suggested that dementia is brought on by the lesion of the left posterior cingulate bundle and retrosplenial cortex causing amnesia by disrupting the cingulate-hippocampal connection or the retrosplenial cortex itself, with the lesion of the left occipitoparietal subcortex causing frontal dysfunction by disrupting the dorsal limbic pathway and occipitofrontal fasciculus of the prefrontal circuits. PMID- 18386632 TI - [Longitudinally extensive spinal cord lesion in a case of Neuro-Behcet disease]. AB - A 56-year-old right-handed man with recurrent orogenital aphtoid ulcers and bilateral uveitis had presented with memory disturbance, dressing apraxia and constructional apraxia at age 53. Neuro-Behcet disease was diagnosed based on pathergy test results and positivity for HLA-B51. Four months after azathioprine was introduced, he presented with subacute spastic paraparesis and urinary retention at age 56. Neurological examination demonstrated hyperreflexia in the lower limbs without pathological reflexes. He also showed memory disturbance, dressing apraxia and constructional apraxia. Spinal cord MRI showed a longitudinally extensive spinal cord lesion (LESCL) from C1 to Th3 with partial gadolinium enhancement from C6 to C8. Brain MRI showed moderate atrophy of the right temporal and parietal lobes without contrast enhanced lesion. There were hyperintense lesions in the pons, bilateral periventricular white matter and right parietal subcortical white matter. Cerebrospinal fluid analysis showed mild lymphocytic pleocytosis. After intravenous methylprednisolone treatment, clinical symptoms largely resolved and the abnormal intensities with contrast enhancement of the cord disappeared. However, higher cortical dysfunctions were not changed. LESCL may reflect inflammatory venous vasculitis with edema extending along the neural fibers since the lesion shows excellent responses to steroid without neurological sequelae. Differential diagnosis of neurological diseases demonstrating LESCL should include Neuro-Behcet disease. PMID- 18386633 TI - [Hyponatremia with somnolence due to indapamide]. AB - We report here an 83-year-old woman presenting with somnolence possibly induced by indapamide. She was diagnosed as having hypertension (180/110 mmHg), and 1 mg/day of indapamide was administered starting in October, 2002. Two months later, she complained of nausea, vomiting, and appetite loss and frequently fell down. On admission, she was hypotensive (90/54 mmHg). Neurologically, she was in a somnolent state (Japan Coma Scale 2-20), and showed brisk deep tendon reflexes of both upper limbs with bilateral Chaddock signs. Laboratory examination showed severe hyponatremia (115 mEq/l) and hypokalaemia (2.8 mEq/l). On brain MR imaging, there were no remarkable abnormalities, except for multiple lacunar infarctions. After the administration of indapamide was discontinued, her consciousness level and serum electrolytes immediately returned to normal levels. After a good effect for stroke prevention was reported, indapamide was widely prescribed in combination with angiotensin-converting enzyme (ACE) inhibitors, or angiotensin II receptor blocker (ARB) among the neurologists. We should keep in mind the risk of hyponatremia and hypokalaemia occurring in patients receiving indapamide, especially elderly women. PMID- 18386634 TI - [Relationship between cardiac 123I-metaiodobenzylguanidine scintigraphy and cardiovascular autonomic function test (standing test) in Parkinson's disease]. AB - We investigated the correlation between results of 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy and those of cardiovascular autonomic function tests in patients with Parkinson's disease (PD). 123I-MIBG myocardial scintigraphy and a 5-minute standing test were performed in 50 patients with PD and in 19 control subjects. The value of the basal plasma noradrenaline (NA) level was used as an index of basal sympathetic nerve activity, and %NA was used to assess the response of sympathetic nerve activity. In addition, the parameters of DeltaBP and DeltaHR were evaluated to assess the autonomic response of the cardiovascular system. A mild, but significant correlation was observed between the myocardium to mediastinum (H/M) ratio and the values of the plasma NA baseline (r = 0.35, p < 0.05 in early image, r = 0.29, p < 0.05 in delay image). No significant correlation was observed between the H/M ratio and the other parameters (%NA, DeltaBP, DeltaHR). These results suggest that 123I-MIBG myocardial scintigraphy may be associated with the basal sympathetic nerve activity, but not with autonomic nervous response of the cardiovascular system in patients with PD. PMID- 18386635 TI - [Massive bleeding from tracheoarterial fistula in an amyotrophic lateral sclerosis patient treated with long-term invasive ventilation: an autopsy case report]. AB - We report a 43-year-old woman with familial amyotrophic lateral sclerosis (FALS) who died of massive bleeding from a tracheoarterial fistula. Four years after the onset of the disease, she received invasive ventilation by tracheostomy because of respiratory failure. Four years and 7 months later, she showed an abrupt hemorrhage from the tracheostomy and died. The postmortem examination revealed a fistulous tract between the tracheal mucosal ulcer and the brachiocephalic trunk. The ulcer was in close proximity to the tracheostomy tube. In order to avoid such unexpected complications, we should observe the contact site between the tracheal mucosa and the tracheal tube in chronic tracheostomy patients. PMID- 18386636 TI - [Influence of connexin 43 gene knockout on the analgesic effect of acupuncture in visceral pain mice]. AB - OBJECTIVE: To explore the possible central and peripheral mechanisms of connexin 43 gene knockout in acupuncture analgesia. METHODS: Fifty-four wide type (WT) mice and 54 connexin 43 gene knockout (heterozygote, HT) mice were randomly divided into WT control group, WT model group, WT acupuncture group, HT control group, HT model group and HT acupuncture group (n = 18/group). Visceral pain model was established by intraperitoneal injection of 0.6% acetic acid (0. 1 mL/10 g). "Zhongwan" (CV 12) and bilateral "Zusanli" (ST 36) were punctured with filiform needles and stimulated for 30 min by manipulating the needle for 30 s every 5 min. The latency and the number of body-writhing response were observed and the contents of beta-endorphin (beta-EP) in hypothalamus and serum prostaglandin E2 (PGE2) were detected with radioimmunoassay (RIA). RESULTS: There was no significant difference in the latency and the number of body-writhing response, and contents of hypothalamic beta-EP and serum PGE2 between HT and WT control groups (P > 0.05). Compared with the corresponding control groups, the latency of body-writhing in WT model and HT model groups shortened significantly and the number of body- writhing increased considerably in two model groups (P < 0.01). While in comparison with WT model group, the latency of body-writhing prolonged significantly and the number of body-writhing decreased apparently in WT acupuncture group (P < 0.01); but no marked differences were found between HT model and HT acupuncture groups in these two indexes (P > 0.05). Compared with the corresponding control groups, the contents of both beta-EP and PGE2 increased obviously in WT model and HT model groups (P < 0.05). In comparison with WT model group, beta-EP levels in WT acupuncture group increased further significantly (P < 0.05), and serum PGE2 in WT acupuncture group decreased obviously (P < 0.05); but no significant changes were found between HT model and HT acupuncture groups in beta-EP and PGE2 levels (P > 0.05). CONCLUSION: Acupuncture has a marked antinociceptive effect in WT mice with visceral pain, which may be related to its effects in increasing hypothalamic beta-EP and decreasing serum PGE2; while in connexin 43 gene knockout mice, all the above-mentioned effects of acupuncture are eliminated, indicating an important role of connexin 43 in the analgesic effect of acupuncture. PMID- 18386637 TI - [Effect of electroacupuncture of local plus distal acupoints in the same segments of spinal cord on spinal substance P expression in rats with chronic radicular pain]. AB - OBJECTIVE: To observe the effect of electroacupuncture (EA) of local plus distal acupoints on spinal substance P expression in chronic radicular pain (CRP) rats so as to investigate its underlying mechanism in the treatment of chronic radical pain. METHODS: Twenty-five Wistar rats were randomized into control, model, local acupoints [LA, bilateral "Jiaji" (EX-B 2)], distal acupoints [DA, bilateral "Yanglingquan" (GB 34)], and LA+ DA groups, with 5 cases in each. CRP model was duplicated by implanting a gel-silicon wrapping the nerve root of L4 under anesthesia. EA (2 Hz, < or =2 mA) was applied to EX-B2 and GB34 for 30 min for 8 days. Pathological changes of the local focus tissue were observed by HE dyeing, and the animals' motor state was also observed. The pain threshold was detected by using tail-flick method. Substance P immunoreactive (IR) positive product of the spinal dorsal horn (L3-L5) was detected by immunohistochemical method and expressed as integrated optical density (IOD). RESULTS: The motor scores on day 35 after modeling and pain threshold values on day 4 and 8 after treatment in EX B2 group, GB34 group and EX-B2 + GB34 group were significantly higher than those in model group (P < 0.05, 0.01). Compared with model group, IOD values of SP in the spinal dorsal horn in EX-B2 , GB34 and EX-B2 + GB34 groups were significantly lower (P < 0.05). There were no significant differences among the 3 EA groups in the expression of SP in the spinal dorsal horn. CONCLUSION: EA of EX-B2, GB34 and EX-B2 + GB34 all has a good analgesic effect in CRP rats, which may be realized partially by suppressing the release of SP in the spinal cord. No significant differences were found among local acupoint, distal acupoint and local plus distal acupoint groups in relieving CRP, improving motor and decreasing SP expression. PMID- 18386638 TI - [Effects of manual acupuncture and electroacupuncture on Ca2+ content and Ca2+ ATPase Activity in sarcoplasmic reticulum of skeletal muscle cells in rats during acute swimming exercise]. AB - OBJECTIVE: To observe the effects of manual acupuncture (MA) and electroacupuncture (EA) on Ca2+ content and Ca2+ -ATPase activity in sarcoplasmic reticulum of skeletal muscle cells in rats of acute swimming exercise (ASE) so as to reveal its underlying mechanism in strengthening motor abilities. METHODS: Sixty male SD rats were randomly divided into control, model, MA and EA groups, with 15 cases in each. Rats in MA group and EA group were treated respectively with MA (120-140 twirlings/min, 30-60 s/time every 3-5 min) and EA (500-800 Hz, 0.20-0.25 mA) stimulation of "Dazhui" (GV 14), "Mingmen" (GV 4), "Huantiao" (GB 30) and "Zusanli" (ST 36) for 20 min before ASE. After ASE (120 min), the rats anesthetized with urethane (1 g/kg) were killed for sampling skeletal muscles (quadriceps muscle of thigh) and for detecting Ca2+ content and Ca2+ -ATPase activity with thymol blue chromatometry and chemical chromatometry respectively. Results In comparison with control group, Ca2+ content and Ca2+ -ATPase activity in sarcoplasmic reticulum of skeletal muscle cells in model group were obviously lower (P < 0.01, 0.05); while compared with model group, both Ca2+ content and Ca2+ -ATPase activity in sarcoplasmic reticulum in MA and EA groups increased significantly (P < 0.01). The activity of Ca2+ -ATPase of MA group was evidently higher than that of control group (P < 0.05). No significant differences were found between MA and EA groups in Ca2+ content and Ca2+ -ATPase activity (P > 0.05). CONCLUSION: Both MA and EA can increase Ca2+ -ATPase activity and Ca2+ content in sarcoplasmic reticulum of skeletal muscle cells in motor fatigue rats, which may contribute to their effects in strengthening motor ability. PMID- 18386639 TI - [Effect of Ca2+ complexation on oxygen partial pressure in the Bladder Meridian in goats]. AB - OBJECTIVE: To explore the relationship between energy metabolism and meridian phenomenon. Methods Ten Boer hybridism goats mitigated under chlorpromazine (0.85 mg/kg, intramuscular injection) were subjected to the present study. Changes of oxygen partial pressure (PO2) in "Ganshu" (BL 18), "Dachangshu" (BL 25) and "Guanyuanshu" (BL 26) of Bladder Meridian and in 3 non-acupoints (3 cm apart from BL18, BL25 and BL26 separately on the unilateral side) were recorded by using a needle-type tissue oxygen tension sensor before and following local injection of EDTA-Na2 (200 microL, 0.05 kg/mol, for complexing Ca2+) and 0.9% saline. RESULTS: (1) Compared with the corresponding non-acupoints, Ca2+ potentials in BL18, BL25 and BL26 were all significantly higher (P < 0.05, 0.01). (2) In comparison with the corresponding non-acupoints, the values of tissue PO2 of BL18 and BL26 were significantly higher (P < 0.01). (3) After injection of EDTA-Na2, PO2 values in both BL18, BL26, and their respective non-acupoints increased significantly in comparison with those of either injection of saline or their respective basal values (P < 0.01, 0.05). Compared with those of their corresponding non-acupoints PO2 values in both BL18 and BL26 increased markedly (P < 0.01). CONCLUSION: Ca2+ potential of the Bladder Meridian was significantly higher than that of non acupoints, and after complexing Ca2+, PO2 in acupoints and non-acupoints increased markedly, suggesting that Ca2+ may play an important role in meridian signaling and production of acupuncture effect. PMID- 18386640 TI - [Effect of acupotomy lysis on SP and CCK-8 contents in hypothalamus and spinal cord in rats with transverse process syndrome of the third lumbar vertebra]. AB - OBJECTIVE: To observe changes of SP and CCK-8 contents in the hypothalamus and spinal cord after performing acupotomy lysis so as to study its central analgesic effect in rats with transverse-process syndrome of the third lumbar vertebra. METHODS: Twenty-eight SD rats were randomly divided into normal control group, model group, acupotomy lysis group and electroacupuncture (EA) group, with 7 cases in each one. The model of the third lumbar vertebra transverse process syndrome was established by inserting a piece of gelatin sponge into the deep subfascial part of the 3rd lumbar transverse process. Acupotomy lysis was conducted 1 time/week for 2 weeks. EA (2/100 Hz, 1-2 mA) was applied to the left "Shenshu" (BL 23)-"Yaoyangguan"(GV 3) for 20 min/time, 3 times/week and for 2 weeks. SP and CCK-8 contents in the hypothalamus and spinal cord were detected with enzyme linked immunosorbent assay (ELISA). RESULTS: In model group, SP and CCK-8 contents in the hypothalamus and spinal cord were increased significantly in comparison with normal control group (P < 0.01). Compared with model group, both SP and CCK-8 contents in hypothalamus and SP content in spinal cord in acupotomy lysis and EA groups were apparently lower (P < 0.01, 0.05). CCK-8 content in spinal cord in cupotomy lysis group was lower markedly than that of model group (P < 0.01), while no significant difference was found between model group and EA group in CCK-8 level in spinal cord (P > 0.05). CONCLUSION: Acupotomy lysis can downregulate SP and CCK-8 contents in hypothalamus and spinal cord, which may contribute to its effect in relieving pain from transverse process syndrome of the third lumbar vertebra. PMID- 18386641 TI - [Effect of transcutaneous acupoint electrical stimulation on blood bioactive compounds involving cerebral injury during craniotomy]. AB - OBJECTIVE: To observe the effect of transcutaneous acupoint electrical stimulation (TAES) on plasma ET, CGRP and serum IL-6, S100beta during craniotomy. METHODS: Fifty patients scheduled for neurosurgery were randomly divided into TAES group (n=25) and control group (n=25). TAES (2/100 Hz, 8-12 mA) of bilateral Hegu (LI 4)-Quchi (LI 11), Zusanli (ST 36)-Sanyinjiao (SP 6) was administrated for patients of TAES group starting 30 min before anesthesia till the end of the operation. Anesthesia of the patients was maintained with sevoflurane inhalation and intermittent intravenous injection of boluses of sulfenany and vecurnium bromide. Jugular venous blood samples were taken at preanesthesia (T0), 1 hour after craniotomy (T1), closure complete (T2), 24 hours (T3) and 48 hours (T4) after operation respectively for detecting contents of plasma endothelin (ET), calcitonin gene-related peptide (CGRP) and serum interleukin (IL)-6, S100beta with radioimmunoassay and enzyme linked immunosorbent assay separately. RESULTS: Compared with control group, plasma ET at T2, serum IL-6 at T1 S100beta at T1 and T4 in TAES group all decreased significantly (P < 0.01, 0.05), while serum IL-6 at T3 increased remarkably (P < 0.05). It suggests that after TAES, CGRP/ET was improved, favoring cerebral microcirculation to reduce surgery-induced cerebral injury. There were no significant differences between two groups at different time courses in plasma CGRP concentrations (P > 0.05). CONCLUSION: TAES can regulate plasma CGRP/ET and serum IL-6, lower serum S100beta level, which may contribute to its effect in relieving craniotomy-induced brain injury. PMID- 18386642 TI - [Electrical stimulation promote proliferation and differentiation of endogenous neural stem cells in normal and injured spinal cord]. AB - This article briefly reviews the general understanding of endogenous neural stem cells (eNSCs) in the CNS over the past 50 years. The proliferation and differentiation of neural progenitors continue throughout the lifetime, instead of cell division stopping at the postnatal period. In the spinal cord, these cells differentiate into different kinds of glial cells. Based on his current research in spinal cord injury (SCI), the author reports that the implantation of functional electrical stimulation (FES) in the motor cortex or the peripheral nerve increases cell birth and differentiation of eNSCs in the spinal cord. Furthermore, electrical stimulation promotes remyelination and neural repair. Finally, this article discusses the possibility of the application of eNSOs and FES in electroacupuncture research. PMID- 18386643 TI - [The messengers from peripheral nervous system to central nervous system: involvement of neurotrophins and cytokines in the mechanisms of acupuncture]. AB - Neurotrophic factors and cytokines are involved in the regulation of neuronal survival, axonal myelination, and synaptic plasticity in both central nervous system (CNS) and peripheral nervous system (PNS). The members of the neurotrophic factor family include nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5). These molecules bind to two types of receptors: (1) tyrosine kinase receptors (TrkA, TrkB, TrkC) and 2) common neurotrophin receptor (p 75 NTR). The internalization and retrograde axonal transport of neurotrophin receptors are important for their signal transduction supporting neuronal survival, synaptic plasticity, and axonal myelination. In addition, a growing body of data suggests that neurotrophins are involved in the pathophysiologicl courses of inflammatory pain, neurodegenerative disease, and psychiatric diseases. Cytokines, including IL-1, IL-2, IL-6, and TNF alpha,are important mediators of the immune response and play a key role in the diseases by acting on inflammatory immune cells, neuronal cells, muscle cells, and vessel cells. Interestingly, some cytokines (e.g. TNF-alpha, IL-2, TGF-beta) are also able to regulate synaptic plasticity and affect CNS functions. The neurotrophins and cytokines release in response to various stimuli, such as electronic stimulation, or inflammation. This crosstalk from PNS to CNS is involved in the pathophysiology of many human diseases and may contribute to the effects of acupuncture. Based on our knowledge to neurotrophins and cytokines, we proposed the neurotrophin/cytokine hypothesis for the mechanism of acupuncture. This hypothesis may initiate the discussion on the possible roles of neurotrophins/cytokines in the therapeutic effects of acupuncture and shed light to the discovery of mechanism of acupuncture in the treatment of devastating diseases. PMID- 18386644 TI - [The pathological neural plasticity and its application in acupuncture research]. AB - The central nervous system (CNS) is involved in a variety of disease conditions. Some seeming peripheral diseases, like chronic pain and disorders in major organs, indeed have clear pathological basis in the CNS. On the other hand, some clinically-beneficial peripheral stimulation, such as acupuncture and massage, exerts significant influence on central neurons. This review attempts to summary recent findings in neuroscience about how pathological insults long-term plastic changes within neural circuits, leading to maladaptive behaviors. This neuroplasticity-based theory not only conceptualizes a cellular mechanism for a plethora of neuropathophysiology but also provides clinical strategies for treating neural diseases. Drug addiction is a chronic brain disease, defined as compulsive drug-seeking, drug-craving, and drug-taking behaviors. Extensive experimental evidence suggests that following exposure to drugs of abuse, neurons within the mesolimbic dopamine system undergo a series of plastic changes that may lead to compulsive emotional and motivational states. It is believed that the first step to unlock the secret of drug addiction is to identify, evaluate, and conceptualize drug-induced neural plasticity. Synaptic plasticity is one form of neuroplasticity that has been best characterized. Using addiction-related synaptic plasticity as a working model, this review attempts to depict the general concept and experimental approach in studying the pathophysiological neural basis of acupuncture. PMID- 18386645 TI - [Biochemical physiology of nitric oxide over acupuncture points and meridians: new approach and perspectives]. AB - We have found that nitric oxide (NO) contents are increased in skin acupoints associated with enhanced neuronal NO synthase (nNOS) protein level in rats. Our recent studies show that L-arginine-derived NO synthesis increases skin electric currents over acupoints in rats. Consistently, norepinephrine (NE) turnover rate is decreased in acupoints with high skin electric currents, and enhanced 3H-NE synthesis/release in acupoints/meridians is facilitated by presence of an exogenous NO donor and inhibited by an inhibitor of NO synthesis. We have developed an innovative method to collect and measure NO metabolites from acupoints and meridians on the skin surface in humans. NO contents are consistently increased over PC and BL acupoints in humans, and increased NO content is reduced by anti-bacteria on skin surface. We conclude that the nonenzymatic reduction of nitrate by bacteria is involved in chemical generation of NO on skin acupoints/meridians in addition to neuronal NOergic system. NO mediates noradrenergic function on skin sympathetic nerve activation, which contributes to low resistance characteristics of acupoints and meridians. PMID- 18386646 TI - [Acupuncture and innate immunity]. AB - Acupuncture is recognized as a procedure to regulate functions of human body. In addition to its known effects on nerve system, emerging evidence suggests that acupuncture also delivers its efficacy through innate immune system. Innate immune system includes three major parts, cytokine activity, complement activity, and cell-based activity. Although initially characterized as natural defense mechanism, innate immune system, especially, the Th1/Th2 cytokine/chemokine networks, plays important roles in regulation of many physiological functions including inflammatory/anti-inflammatory activity, pain, metabolism, cell proliferation and apoptosis. Moreover, innate immune system interacts closely with peripheral and central nerve systems. Roles of cytokines/immune cells (macrophages and T cells) in nerve functions and pathogenesis of neuronal diseases have been increasingly recognized. Likewise. ONS through hypothalamic pituitary-adrenal (HPA) axis and cholinergic anti-inflammatory pathway adjusts immune activity. We present a testable model to validate the effect of acupuncture in innate immune system. The model emphasizes the existence of dynamic cytokine network. The goal is to demonstrate that the combined neuro immune function is a general mechanism of acupuncture in the treatment of different diseases. PMID- 18386647 TI - [Acupuncture clinical studies and evidence-based medicine--an update]. AB - Acupuncture has been widely used in the West in recent years and demand has been growing for scientific evaluation of its clinical efficacy. The practice of evidence-based medicine has brought new challenges in the design of acupuncture research, and publication of randomized clinical trials on acupuncture has significantly increased. While systematic reviews of these trials have advanced our current knowledge, they have exposed deficiencies in research design and revealed that one design can not answer all research questions. Few clinical studies conducted in China have been published in the West, and most published in Chinese suffer from methodological design flaws that render the results unreliable and unconvincing. Such flaws include inadequate or no randomization, inadequate control, unsatisfactory outcome measurements, lack of proper concealment, insufficient follow-up, and improper statistical analysis. To foster high quality acupuncture clinical research in China, we must cultivate innovation and creativity in research design. It is unwise to simply follow or copy the research methodology of Western pharmaceutical studies. Acupuncture and traditional Chinese medicine (TCM) must be evaluated using rigorous scientific methods that preserve the essence of TCM concepts, so that acupuncture and TCM, these ancient healing arts, can continue to play an important role in the health care systems of modern societies. PMID- 18386648 TI - [Current situations about studies on the evaluation of clinical efficacy of acupuncture]. AB - To evaluate the efficacy of acupuncture scientifically and objectively is a major issue for the development of acupuncture medicine. Although some progresses have already been gained in the research of the evaluation of clinical therapeutic effects of acupuncture at present, some problems as empirical evaluation method, following evaluation indexes of Western medicine, poorer quality of evaluation studies, etc. still exist. Thus, it is urgent to establish a reasonable evaluating system for acupuncture clinical efficacy which accords with the theory of acupuncture, embodies the characteristics and advantages of acupuncture clinic, perfecting the clinical evaluation method, and improving the quality of clinical efficacy evaluation and accelerating the sustainable development of acupuncture clinic. Through reasonable analysis on the current situations about the clinical evaluation research of acupuncture and combining some new concepts and methods, the authors of the present paper hold that the new evaluation system should embody the characteristic of the theory and clinical practice of acupuncture and fully make use of the methods of evidence-based medicine and some measuring scales. They also advocate trying to establish an "evidence-based-goal attainment scale" method by combining standardized population evaluation with individualized diagnosis-treatment evaluation so as to enhance the international development of acupuncture. PMID- 18386649 TI - [Characteristics of peripheral afferent nerve discharges evoked by manual acupuncture and electroacupuncture of "Zusanli" (ST 36) in rats]. AB - OBJECTIVE: To observe the nerve discharge changes responding to stimulation of "Zusanli" (ST 36) by manual acupuncture (MA) and electroacupuncture (EA), so as to reveal the changing regularity of peripheral nerve response. METHODS: Thirty male SD rats were used in the present study. After anesthetizing with urethane (75 mg/kg) and dissecting a branch of the sciatic nerve innervating ST36 region in the right hind-limb, the ipsilateral ST36 was stimulated with MA (by lifting and thrusting and rotating the acupuncture needle, 1 manipulation/s) and EA (5 V, 10 V, 15 V, and 3 Hz). Discharges of the nerve tract was recorded for 1 min before, during and after MA or EA stimulation respectively, and analyzed in spike counts. RESULTS: After lifting, thrusting and rotating the needle, the nerve discharge increased to (74.20 +/- 18.92) spikes/s from (42.20 +/- 1.98) spikes/s of pre-MA (P < 0.05), then turned to (41.40 +/- 14.85) spikes/s immediately after cease of MA. After manual lifting and thrusting the needle, the nerve discharge increased to (60.60 +/- 10.79) spikes/s from (42.00 +/- 3.89) spikes/s before MA (P < 0.05), and returned to (45.00 +/- 18.10) spikes/s immediately after cease of MA. No changes were found after manually rotating the needle (P > 0.05). EA at ST36 by increasing the intensity to 5 V began to elicit similar changes of nerve discharges, and increased significantly in a voltage-dependent manner within 5-15 V. EA at the strength of 15 V could produce a marked increase of nerve discharge, being increased to (64.20 +/- 9.36) spikes/s from (42.60 +/- 3.87) spikes/s before EA (P < 0.05), and then, returned to (48.00 +/- 7.55) spikes/s immediately after cease of EA. CONCLUSION: MA of lifting and thrusting, and particularly that of rotating combined with lifting and thrusting at ST36 can significantly potentiate the discharge activity of the sciatic nerve tract, with the response of the latter being more apparent. EA of ST36 at the strength of 10-15 V has a similar effect. It suggests that both mechanical and EA stimulation of ST36 at a certain intensity can produce a similar change of discharges of the afferent nerve innervating ST36 in the rat. PMID- 18386650 TI - ACCESS Healthcare Coalition makes inroads on Medicaid funding crisis. PMID- 18386651 TI - Hospitals must lead in reform discussions. PMID- 18386652 TI - Medicaid mess a matter of access. PMID- 18386653 TI - The age of certainty. PMID- 18386654 TI - Unconscious fantasy and disturbances of conscious experience.1969. PMID- 18386655 TI - Ubiquitous daydreams and unconscious fantasy: a reassessment of Arlow's "Unconscious fantasy and disturbances of conscious experience". PMID- 18386656 TI - Two readings of Arlow's "Unconscious fantasy and disturbances of conscious experience": one old and one "green". PMID- 18386657 TI - The working alliance and the transference neurosis. 1965. PMID- 18386658 TI - Commentary on Greenson's "The working alliance and the transference neurosis". PMID- 18386659 TI - Discussion of "The working alliance and the transference neurosis," by Ralph R. Greenson. PMID- 18386660 TI - The mind: psychoanalytic understanding then and now. AB - The author discusses the evolution of psychoanalytic understanding from Freud's time to the present, citing the influence of various sociocultural changes. He addresses Freud's proper place in history and notes ways in which Freud's contributions cast him as belonging to Romanticism. Freud's shift from the topographic model of the mind to the structural one, and the influence of this on psychoanalysis, is discussed, as well as important developments in the field since Freud. The author focuses particularly on difficulties encountered in psychoanalytic practice today, and he describes what he has termed organizing interpretations as uniquely valuable in the treatment setting. PMID- 18386661 TI - On knowing oneself directly and through others. AB - For certain patients, the experience of self lacks solidity and conviction. These patients appear to be unable to know themselves directly and turn to others in their environment and in fantasy in order to discover or confirm a vision of themselves. The author argues that we can come to understand these patients' shadowy sense of self by looking at direct self-knowledge and knowledge of the self that is acquired through others as two storylines. Detailed clinical material from the analysis of a woman who came to analysis feeling shadowy and insubstantial illustrates the value of attention to both storylines and to the changing relationship between them. PMID- 18386662 TI - Vicious circles of punishment: a reading of Melanie Klein's Envy and gratitude. AB - Staged as a moral drama, Melanie Klein's Envy and Gratitude (1957) straddles two eras: one reflecting a simpler more concrete, historical method, and the other an approach of greater complexity, focused on the transferential moment. Embedded within the transference are the vicious circles of envy and punishment that are the hallmark of Klein's text. Punishment itself which is always punishment of both self and other, creates its own vicious circle, set in motion by a single act. In the sadomasochistic enactments that result, the moral force of Klein's categories fuels the countertransference. A detailed clinical hour is presented. PMID- 18386663 TI - Id analysis and technical approaches. AB - The author argues that the technical advances stemming from Freud's (1923) introduction of the structural theory permit a more naturalistic and specific approach to analyzing unconscious conflict, thus facilitating id analysis. The earlier topographical technique underestimated the role of suggestion; often, it entailed interference with patients' capacity for self-observation, as well as with the exploration of their own drive derivatives. In order to illustrate the type of id material obtainable with a contemporary ego psychology approach, the author presents clinical vignettes and commentaries. It is recognized that clarifications, defense interpretations, and Gray's close-process interventions may need to be adapted to different cultural milieus. PMID- 18386664 TI - A measure of agreement: an exploration of the relationship of D. W. Winnicott and Phyllis Greenacre. AB - The British psychoanalyst D. W. Winnicott and the American psychoanalyst Phyllis Greenacre were both deeply absorbed by the vicissitudes of the infant's and young child's psychic development. Their clinical observations and theoretical ideas display striking convergences and reciprocal influences. Winnicott was deeply influenced by Greenacre's account of maturational processes, an important stimulus to his thinking that originated outside of the British Psychoanalytical Society. Greenacre's writings on early ego development and creativity were influenced by Winnicott's concept of transitional phenomena. The fact that these relationships have remained unexplored until now indicates the need for less insular accounts of the development of psychoanalytic thought on the two sides of the Atlantic. PMID- 18386665 TI - Touched by Grace during the psychoanalytic hour: the transformation of a religious resistance. AB - Case material is presented illustrating a phase in religious transformation in which the quality of the religious moment -first expressed in transference hints and a dream, and finally augmented by an idiosyncratic enactment of the patient's became sufficiently intense that it crossed formal religious boundaries. The patient resisted direct reference to her religious beliefs, yet the deeper roots of her God representations took alternative forms of expression. The analyst's appreciation of this, which was rendered articulable through a carefully refined countertransference experience, eventually enabled a sincere experience of joining, one that superseded apparent religious differences between analyst and patient. PMID- 18386666 TI - A note on the use of the concept of the soul in psychoanalytic discourse. PMID- 18386667 TI - [Patient education and illiteracy in Switzerland]. PMID- 18386668 TI - [What is the role for the non-specialist physician in the care of patients with hemophilia?]. AB - Haemophilia is a relative rare disorder and often poorly known by physicians who are reluctant to take care of a disease with emotional aspects. However, some types of haemophilia (mild haemophilia) are diagnosed late in life, for example at the time of a haemorrhage after surgery. Moreover, the care of a person with haemophilia (PAH) is not anymore the prerogative of haemophilia centres, especially at the time when prophylaxis is more often prescribed. Indeed, thanks to preventive treatment, PAH bleed less but they may have other health problems encountered by primary care physicians or specialists. Therefore physicians should know how to recognize this coagulation disorder and how to manage these patients. It is still important that treating physicians and PAH remain in contact with treatment centres. PMID- 18386669 TI - [Vascular diseases: what screening could be done in the office?]. AB - The screening of vascular pathologies in physician offices starts with precise medical history and clinical exam. Tools like the Edinburgh Claudication Questionnaire for the peripheral artery disease or the Wells score for the probability of a thromboembolic event are useful. The measure of the ankle brachial index, the D-dimers or any other biological screening are complementary. In the presence of pathological features, it is recommended to organise a specialised consultation in order to precise diagnosis, treatment and follow-up. The screening of a vascular disease is interesting not only for the management of local symptoms, but also for the associated systemic pathologies to provide a preventive medicine of good quality. PMID- 18386670 TI - [Long term oral anticoagulation: indications and problems]. AB - There are several well established indications for long term oral anticoagulation, including atrial fibrillation, valvular heart disease and some categories of venous thromboembolic disease. Bleeding is the major complication. Apart from the estimated benefits and risks of long term oral anticoagulation in an individual patient, many practical issues interfering with the management of such treatment should be taken into account. These include compliance, factors causing high INR variability and necessity of frequent monitoring. Involving patients in the decision and treatment management highly improves efficacy and security of long term oral anticoagulation. PMID- 18386671 TI - [Seventy years later: prothrombin time revisited]. AB - Introduced by Armand Quick in 1935, the prothrombin time is the test the most frequently performed in haemostasis laboratories. The diversity of origin and purity of the tissue factor contained in the thromboplastin reagents as well as the composition of phospholipids explain the differences in sensitivity to factor(s) deficiency(ies) observed. Unfortunately, the expression of assay results in non anticoagulated patients varies depending on the place the test is performed. For patients under oral anticoagulation treatment, huge standardisation efforts have led to the implementation of the International Normalized Ratio (INR) for the monitoring of the treatment, which has considerably reduced the differences between thromboplastin reagents. It is hoped that a comparable degree of standardization will be achieved for samples from patients with hepatic disease. PMID- 18386672 TI - [Is computed tomographic venography of lower limbs useful in suspected pulmonary embolism?]. AB - Computed tomography (CT) is widely used in diagnostic strategies for suspected pulmonary embolism (PE). As single-slice computed tomography has a limited sensitivity of about 70%, it has to be used in association with lower limbs veins compression ultrasonography (CUS). To improve the test, it has been proposed to add lower extremity CT venography to visualise lower limbs veins. Diagnostic performances of CT venous angiography and CUS have never been compared and there is no published well conducted study suggesting that adding CT venous angiography is useful. Recent studies have confirmed that CUS is no more mandatory when using the recent sensitive multi-slice computed tomography. These data limit the potential use of CT venography of the lower limbs veins in patients with suspected PE. PMID- 18386673 TI - [Frequently asked questions on clopidogrel treatment: indications, resistance, and biological evaluation in vascular patients]. AB - Clopidogrel is the most widely used thienopyridine in cardiovascular patients. Its indications include acute coronary syndromes and stenting, in association with aspirin. In stable atherothrombotic patients, clopidogrel is a valuable alternative to aspirin. Studies have shown an association between the biological response to a loading dose of clopidogrel and clinical events after stenting. This variability of the biological response is probably multifactorial (compliance, genetics, weight, drug interactions). Several aspects, such as the lack of standardized tests, of validated cut-offs to define resistance and of alternative treatments with proven efficacy in case of a biological resistances, do not currently allow performing these tests in everyday care and limit their use in the clinical research setting. PMID- 18386674 TI - [Metabolic syndrome: jumble syndrome of obesity or specific entity? Which treatment: diet or polypill?]. AB - The reality of metabolic syndrome (MS) as a specific entity is debatable. However, the simple measure of waist circumference (>94 cm in men and >80 cm in women) is useful: (1) to check for insulin resistance by measuring serum levels of fasted glucose and insuline, cholesterol, triglycerides; (2) to look for diseases associated with MS such as hypertension, non alcohoolic steatohepatitis, sleep apnea, polycystic ovary disease, hypogonadism and to measure serum levels of ferritine, ALAT, ASAT, urate acid, CRP hs, testosterone and (3) to make obese people aware of their risk of becoming diabetic and to motivate them to change their life style. The utility of exercise and of various diets is discussed as well as the efficiency of drugs acting on different components of MS such as rimonabant, orlistat, metformin, glitazones, telmisartan and testosterone. The importance of political measures to fight the obesity epidemic is underlined. PMID- 18386675 TI - [Assisted reproduction: data from France]. PMID- 18386676 TI - [Should we place babies in front of the television? (1)]. PMID- 18386677 TI - [In our day, the creators are all around us]. PMID- 18386678 TI - [Physicians and death]. PMID- 18386679 TI - [Readiness to interact]. PMID- 18386680 TI - [Patient advance directives in the tension field between euthanasia and palliative care: "there is no simple solution"]. PMID- 18386681 TI - [Congress on the topic "Nursing diagnosis--introduction and implementation". Promoting more quality in the nursing process]. PMID- 18386682 TI - [Abused and neglected children: less activism, more evidence based management]. PMID- 18386683 TI - [Quality of life and satisfaction of patients with dementia. "We can prove the value of good nursing care" (interview by Katrin Balzer)]. PMID- 18386684 TI - [Competence groups of patients with dementia: directing attention to resources]. PMID- 18386685 TI - [Development of a nursing and care concept for patients with dementia: guide show the way experienced concept]. PMID- 18386686 TI - [Communication and conflict discussions: from "I" to "you"]]. PMID- 18386687 TI - [Pressure ulcers: new data about risk, prevalence and places of development. Results of a study in German nursing homes and hospitals]. AB - In the spring of 2007 the "Institut fur Medizin-/Pflegepadagogik und Pflegewissenschaft der Charite-Universitatsmedizin Berlin" conducted a nationwide prevalence study. Data were collected concerning pressure ulcer risk and pressure ulcer prevalence. In addition, data were gathered about place of development, prevalence rates for various clinical specialties within hospitals, and pressure ulcer location. Altogether, data from 6473 patients and residents were collected and analysed. In nursing homes the percentage of persons at risk for pressure ulcer development was 61.4 percent, in hospitals 38.0 percent. The prevalence of pressure ulcers grade 1 to 4 was 8.4 percent in nursing homes and 15.7 percent in hospitals. In nursing homes as well as in hospitals more than half of the pressure ulcers emerged within the same facilities. For 22.6 percent of pressure ulcers in hospitals the place of development remained unknown. In nursing homes this proportion was 5.6 percent. PMID- 18386688 TI - [Translation of the "Autar Deep Vein Thrombosis Scale" to assess the risk of thrombosis]. AB - At the moment in Austrian hospitals there is no valid and reliable scale available to assess the deep vein thrombosis risk. An alternative method for development of a German instrument of its own is the systematic translation of a standardized instrument in a foreign language which has been proved for its validity. For the translation the English Autar DVT risk assessment scale has been selected. The translation is based on a specific procedure. The results of the first part of the study show that the German version of the Autar DVT risk assessment scale (Autar DVT Scale-D) is a comprehensible and valid instrument as regards to content validity. in the second part further test-theoretic quality criteria of the Autar DVT Scale-D as reliability, validity and practicability will be tested. PMID- 18386689 TI - [Step by step to nursing competence--10: Prevention of thrombosis: physical activity as the best prevention]. PMID- 18386690 TI - [Effectiveness of termination for suspected wrong doing: not valid without a hearing with the employee]. PMID- 18386691 TI - [From case to case: "Mrs. L. is 'odd'"]]. PMID- 18386692 TI - [Recognizing and understanding disorders: Thoracic pain as the leading symptom]. PMID- 18386693 TI - Lifelong learning in nursing: a drilldown of the Macy Foundation report. PMID- 18386694 TI - Recommendations for improving health care through lifelong learning. AB - This column offers reflections on potential operational implications for continuing nursing education providers based on the summary report of the Josiah Macy, Jr. Foundation included in this issue. PMID- 18386695 TI - Autism: part II. Genetics, diagnosis, and treatment. AB - Due to the increasing prevalence of autism, nurses need to become more informed about the disease. This column reviews the genetics, diagnosis, and treatment of autism. PMID- 18386696 TI - Appreciative inquiry. AB - Appreciative inquiry, which emphasizes using affirmative questions, can help create a positive environment. PMID- 18386697 TI - Increasing understanding of nursing research for general duty nurses: an experiential strategy. AB - Misconceptions and trepidation about research abound among practicing nurses. However, in light of the movement toward increasing accountability to consumers and the concurrent drive toward evidence-based practice, the need for nursing research can no longer be ignored. Innovative approaches to augment nurses' training and education in research and evidence-based practice must be incorporated into continuing education programs. The Nursing Research and Evidence-Based Practice Committee of a large tertiary care teaching hospital in Winnipeg, Manitoba, Canada, developed a series of opportunities for staff nurses to participate in research projects and have ongoing exposure to the steps in the research process. The Great Canadian Cookie Experiment was an opportunity to participate in quantitative research. Qualitative data from patients' thank you cards were analyzed in an interactive fashion during luncheon seminars held during Nursing Week in 2 subsequent years. A survey of nurses who participated in the luncheon seminars indicated an overall increase in their knowledge about qualitative research methods and an appreciation for participating in the process of nursing research. Continued visibility of nursing research will contribute to changing nurses' attitudes toward fostering an evidence-based approach to clinical practice. PMID- 18386698 TI - Chairman's summary of the conference. Continuing education in the health professions: improving healthcare through lifelong learning. PMID- 18386699 TI - Concept mapping: an educational strategy to improve graduate nurses' critical thinking skills during a hospital orientation program. AB - Novice graduate nurses (GNs) are often deficient in the critical thinking skills needed to assess and handle patient problems and life-threatening situations. Concept mapping is an effective teaching strategy for developing critical thinking and clinical decision-making skills. This small descriptive comparison study was undertaken to determine whether concept mapping improved critical thinking skills in GNs during a hospital orientation program. Schuster's (2002) Concept Map Care Plan Evaluation Tool was adapted to measure GNs' critical thinking in concept maps based on case studies at the beginning and end of orientation. A paired t test was computed on pre- and post-mean map scores of 14.071 and 16.428, respectively. Concept mapping was a valuable teaching and evaluation strategy for this group that could be used by nursing educators to improve critical thinking and identify and correct areas of theoretical and clinical deficiency. PMID- 18386700 TI - Caseload management skills for improved efficiency. AB - This article provides information about caseload management, which was one of a group of six competencies identified by nurse administrators as needed by new baccalaureate graduates. Caseload management is an important skill for nurses who work with caseloads of patients or clients (e.g., home health nurses, public health nurses, case managers, ambulatory care nurses). Because inadequate information about caseload management is contained in community health nursing textbooks, continuing education and staff development programs need to include caseload management skills to improve the efficiency and quality of nursing care. PMID- 18386701 TI - A study of critical thinking, teacher-student interaction, and discipline specific writing in an online educational setting for registered nurses. AB - Based on work conducted by Laurentian University's School of Nursing and Centre for Continuing Education in Sudbury, Ontario, Canada, working in conjunction with community partners, this article looks at the findings of an analysis of nurses' writing activity in a university-level web-based module for evidence of critical thinking using Johns' Model of Structured Reflection (1995). Also considered are student-teacher interactions and discipline-specific writing. The findings suggest that high levels of critical thinking by nurse learners and growth in thinking and writing competence over time can occur in an online setting. Further highlighted are the role of the instructor, assignment design, and support in fostering such development. PMID- 18386702 TI - Evaluating program effectiveness: creating a reliable and valid tool. AB - This study was done to assess the reliability of a pretest/ posttest design for a mass-casualty burn disaster continuing education (CE) program. Based on program objectives, an instrument to assess knowledge, skills, confidence, and competence in burn assessment and management was designed. The instrument's internal consistency, split-half reliability, and individual items were analyzed. Measures of reliability were moderately high, and item analysis identified no poorly constructed items. The instrument appears to be a reliable measurement of effectiveness for the CE program and will be consistent in measuring changes based on the training. The results may be useful to others who are planning CE training programs. PMID- 18386703 TI - Prevalence and risk factors of urinary incontinence in Khon Kaen menopausal women. AB - OBJECTIVE: To determine the prevalence and risk factors of urinary incontinence in menopausal women. STUDY DESIGN: Prospective Descriptive study. SETTING: Menopausal clinic and gynecological outpatient unit, Department of Obstetrics and Gynecology, Srinagarind Hospital, Faculty of Medicine Khon Kaen University. STUDY POPULATION: Menopausal women attending the menopausal clinic and gynecological outpatient unit. MATERIAL AND METHOD: Between April 23 and July 15, 2004, 229 menopausal women were interviewed by pre-validated questionnaire and in-depth interview to determine the prevalence and risk factors of urinary incontinence. RESULTS: The average age of the study group was 55.8 years. The prevalence of urinary incontinence was 38.86%, the most common type was mixed urinary incontinence (82.02%) followed by stress urinary incontinence (12.36%), and urge urinary incontinence (5.62%). The risk factors of urinary incontinence were vaginal delivery, menopause before 50 years old, and postmenopausal women who had never used hormone therapy. CONCLUSION: The prevalence of urinary incontinence in menopausal women was 38.86%. Mixed urinary incontinence was the most common type. Associated risk factors were vaginal delivery, menopause before 50 years old, and postmenopausal women who had never used hormone therapy. PMID- 18386704 TI - A population-based cohort study of effect of maternal risk factors on low birthweight in Thailand. AB - OBJECTIVES: To determine the maternal risk factors of low birthweight (LBW) in Thailand and to address the possible activities to reduce the incidence of LBW. MATERIAL AND METHOD: The data were obtained from the Prospective Cohort Study in Thai Children (PCTC). Three thousand five hundred twenty two pregnancies initiated the follow-up in the year 2000 at four districts across Thailand The birthweight was retrieved from the Delivery Summary Sheet of the hospitals. The babies born in the hospital were weighed within the first day of life. The babies born at home were weighed within 3 days after birth by the research assistants. Only singleton live births were included in the present study. RESULTS: Three thousand three hundred twenty two live births with birthweight data, 8.6% were LBW. Maternal factors affecting LBW with high attributable fraction (AF) and moderate population attributable risk (PAR) were maternal extreme age (AF = 45.96, PAR = 16.24) and weight gain during pregnancy less than 10 kg. (AF = 40.12, PAR = 16.05). The factors with moderate AF and PAR were first and more than two parities (AF = 21.9, PAR = 15.51) and less consultation to health personnel (AF = 20.96, PAR = 16.98). CONCLUSIONS: Improving nutritional status of pregnant women is a potential activity to reduce the incidence of LBW. Pregnant women with extreme age, first and more than parity two and less consultation to health personnel should also be closely followed-up. PMID- 18386705 TI - Cost analysis of hematopoietic stem cell transplantation in adult patients with acute myeloid leukemia at King Chulalongkorn Memorial Hospital. AB - The purpose of the study was to analyze the first-year cost ofhematopoietic stem cell transplantation (HSCT) program for the treatment of adult patients with acute myeloid leukemia (AML) at King Chulalongkorn Memorial Hospital (KCMH). The present retrospective study was carried out on 67 AML patients treated with bone marrow transplantation (BMT) or peripheral blood stem cell transplantation (PBSCT) at KCMH during the period of 1994 to 2005. The actual total one-year cost from the provider perspective were determined by the reviewing medical records for medical care costs (MCCs) and by adjusting data from the reports of annual cost analysis of KCMH for routine services costs (RSCs). All costs were converted to 2006 values using the Thai consumer price indices. It was found that the full cost of allogeneic HSCT (allo-HSCT) and autologous HSCT (auto-HSCT) in the first year of the program was $22,592.85 and $24,171.25 per case respectively. Cost effective appraisal, comparing with chemotherapy, need to be studied further. PMID- 18386706 TI - Hemoglobin response and influence on left ventricular hypertrophy after 24-week treatment of a biosimilar epoetin-alfa in hemodialysis patients with anemia. AB - BACKGROUND: Anemia is common in end-stage renal disease (ESRD) patients and an important determinant for left ventricular hypertrophy (LVH) in dialysis patients. There are increasing numbers of biosimilar epoetin-alfa entering Thailand. OBJECTIVE: To conduct a prospective trial to evaluate the efficacy and safety of a biosimilar epoetin-alfa (epoetin) (Espogen) in ESRD patients receiving chronic hemodialysis complicated by anemia and to address its impact on the left ventricular mass index (LVMI) and volume index (LVVI) in these patients. MATERIAL AND METHOD: Twenty-two hemodialysis (HD) subjects were recruited from Rajavithi and Huachiew Hospitals. Inclusion criteria were chronic HD, hemoglobin (Hb) < 10 g/dL without preceding treatment (epoetin or transfusion) for 1 month. Echocardiographic baselines were obtained Epoetin-alfa was initially given 4,000 IU subcutaneously twice a week and titrated biweekly to keep the Hb range of 11 to 12 g/dL (titration period 12 weeks). Treatment continued until the end of 24 weeks. Records were made for conventional blood tests, blood pressure, amount of drugs needed to control blood pressure, and adverse events. Echocardiogram was repeated (on observer blinding) at the completion of the present study. RESULTS: After 24-week of epoetin therapy, the predialysis Hb level increased significantly from 8.0 +/- 1.3 g/dL to 11.0 +/- 1.1 g/dL (p < 0.001). The mean dose of epoetin at the present study entry was 143.6 +/- 87.8 IU/kg/ week. At the present study entry, LVH was present in 86.4% of the patients. At the completion of the present study, a decrease in LVMI was observed in 50% of the patients; however, the mean LVMI change was not significantly different. Notably, there were minimal but significant changes in LVEDD (52.8 +/- 7.0 vs. 50.1 +/- 6.9 mm, p < 0.05), LVVI (86.2 +/- 25.2 vs. 75.5 +/- 19.5 mL/m2, p < 0.05) and when subjects were partitioned into tertiles of baseline LVMI, the LVVI change was confined to the highest tertile (103.7 +/- 25.2 vs. 79.6 +/- 21.9 mL/ m2, p < 0.05). The aortic root diameter also significantly decreased despite some increase in blood pressures but without significant change in number of antihypertensive agents. No serious adverse event was observed during the present study period. CONCLUSION: The efficacy of anemia treatment and safety of the biosimilar epoetin-alfa was demonstrated in hemodialysis patients. Significant regression of LVVI and some reduction in LVMI were shown in this 24-week prospective trial. PMID- 18386707 TI - Importance of delayed perfusion with primary angioplasty on short-term mortality in acute myocardial infarction patients. AB - BACKGROUND: Early primary coronary interventions (PCI) in acute ST elevation myocardial infarction (STEMI) is associated with improved outcome and mortality rate but delayed reperfusion especially after 6 hours is still doubtful in terms of clinical benefits because most myocardial muscle are infarcted after 6 hours of onset of chest pain. OBJECTIVE: The aim of the present study was to compare the mortality rate of patients treated with PCI within 6 hours of symptom onset to those treated between 6 to 24 hours after the onset of STEMI. MATERIAL AND METHOD: The present study included consecutive patients from the data of the Fast Track Registry of King Chulalongkorn Hospital from June 1, 1999 to October 31, 2003 to compare the thirty-day mortality of patients treated with early or delayed PCI (0-6 hours vs. 6-24 hours after symptom of chest pain) for STEMI. RESULTS: Two hundred and sixteen patients who underwent PCI were enrolled. Male gender (82% vs. 64.9%, p = 0.03) and history of smoking (72.1% vs. 50%, p = 0.04) were predominant in the early treatment group (ETG) vs. the delayed treatment group (DTG). Mean age (60.5% vs. 61.03%, p = 0.11), diabetes (31.4% vs. 29.7%, p = 0.82), hypertension (64.0% vs. 54.1%, p = 0.20), dyslipidemia (58.1% vs. 60.8%, p = 0.73), and ejection fraction < 40% (22.8% vs. 32.0%, p = 0.625) were similar in both groups. There were no differences in angiographic finding and hospital management. Door to balloon and total delay time were 124.13 +/- 143.27 min and 407.94 +/- 268.183 min, respectively. The thirty-day mortality (9.01% vs. 12.76%, p = 0.379) and I year mortality (12.4% vs. 16 9%, p = 0.532) were not significantly determined by Log rank test in both groups. As for cardiogenic shock, ETG tended to have a lower thirty-day mortality than DTG but no statistically significant difference (12.5% vs. 50.0%, p = 0.0809). CONCLUSION: The delayed PCI up to 24 hours in STEMI does not increase short-term mortality at thirty days; therefore, it may still have benefit in STEMI patients. However it tended to have higher short-term mortality than early PCI especially in cardiogenic shock but showed no statistical significance. PMID- 18386708 TI - Very high dose phenobarbital for refractory status epilepticus. AB - BACKGROUND: Refractory status epilepticus (RSE), defined as status epilepticus that fails to respond to first, second and third-line therapy. The RSE is associated with high morbidity and mortality. Treatment guidelines of RSE give a spectrum of options, such as, continuous intravenous (i.v.) midazolam (MDL), or continuous i.v. propofol (PRO) as alternatives to phenobarbital (PB) or continuous i.v. pentobarbital (PTB). OBJECTIVE: To study the efficacy of very high-dose phenobarbital (VHDPB) for treatment RSE. STUDY DESIGN: Retrospective study MATERIAL AND METHOD: The authors collected and analyzed data from adult patients who were diagnosed with RSE. RESULTS: The authors present 10 patients with RSE who were treated with VHDPB. All of them were generalized convulsive status epilepticus (GCSE). Ages ranged from 16-86 years old (mean.: 43 years). PB dosage ranged 40-140 mg/kg/day (mean: 70 mg/kg/day). The duration of status epilepticus (SE) varied widely, ranged 1-44 days (mean: 7 days). PB level ranged 35.29-218.34 ug/mL (mean 88.1 ug/mL). RSE was controlled by VHDPB 70%, 30% were not controlled. CONCLUSION: VHDPB were considered as alternative treatment for RSE. PMID- 18386709 TI - Efficacy and safety of gentamicin by interval and intravenous dosage adjustment based on the gestational age in Thai neonates. AB - OBJECTIVE: To determine the pharmacological efficacy and safety of the gentamicin regimen that adjusts intravenous dose and interval based on the gestational age (GA) in Thai neonates. MATERIAL AND METHOD: Neonates aged < or = 7 days, who had received gentamicin for clinically suspected or high risk of sepsis and had no contraindication to gentamicin usage were enrolled. They were stratified into four groups by GA as < or = 29, 30-33, 34-37 and > or = 38 weeks gestation. Gentamicin administration in each group was 5, 4.5, 4 and 4 mg/kg/dose every 48, 36, 36 and 24 hours respectively according to Neofax regimen. Peak serum gentamicin concentration (SGC), trough SGC and serum creatinine (Cr) were obtained. RESULTS: Forty-nine neonates were enrolled. Forty-four (89.7%) had peak SGC within the desirable range (5-12 mg/L). Three neonates had slightly high peak SGC. Their peak SGCs were 13.0, 12.21 and 12.20 mg/L. Two neonates had slightly low peak SGC. Their peak SGCs were 4.91 and 4.4 mg/L. All neonates had trough SGC below 2 mg/L. None had significant rising of serum Cr during the present study period. CONCLUSION: This gentamicin regimen yielded good pharmacological efficacy and safety in Thai neonates, who were in the first week of life and had no renal function impairment. PMID- 18386710 TI - Physical effects of sexually abused children and adolescents at Taksin Hospital. AB - OBJECTIVE: To study the incidences and physical effects of sexually abused children and adolescents. MATERIAL AND METHOD: Rape records and records of 250 sexually abused children and adolescents treated at Taksin Hospital between January 1, 1999 and December 31, 2004 from Child-Women Protection Center were studied retrospectively. RESULTS: The incidence of sexual abuse in children and adolescents was 4.74 per 10,000 cases of age-adjusted patients at the OPD and 22.97 at the emergency department. Most of the cases (97.20%) were females. Mean age was 13.74 +/- 4.27 (2-20) years old. Most of the cases were early adolescents (52.40%), late adolescents 32.40%, and children 15.20%. Thirty-nine cases (15.60%) had physical injuries, 36 cases (14.40%) had external genital injuries, 25 cases (10.00%) had gonococcal infections, 15 cases (6.20%) had bleeding in the vaginal canal, and eight cases (3.20%) were pregnant. Two hundred and twenty six cases (90.40%) were reported to the police. There was correlation between age group and hymen tearing (p-value < 0.001), gonococcal infection (p-value < 0.01) and sperm finding (p-value < 0.001). However there was no correlation between age group and gender, physical injury, genital injury, bleeding in the vagina and acid phosphatase finding. In addition, there was correlation between physical injury and bleeding in the vagina (p-value < 0.01) but no correlation between genital injury and hymen findings. CONCLUSION: Sexual abused victims need immediate attention for the traumatic impacts of their physical, psychological, and emotional conditions, as well as on their social impact. Thus, it is imperative that protection be exerted over the treatment to prevent recurrence. PMID- 18386711 TI - A survey of coagulation laboratory practice in Thailand: the first step to establish a National External Quality Assessment Scheme (NEQAS) for blood coagulation. AB - BACKGROUND: External quality assessment (EQA) is an essential component of laboratory quality assurance. In Thailand, there is no EQA program for coagulation tests at the national level. OBJECTIVE: To collect the necessary data in the first step to set up a National External Quality Assessment Scheme (NEQAS) and to assess the status of coagulation laboratory practice in Thailand. MATERIAL AND METHOD: Questionnaires were sent to hospitals to obtain information about the hospitals, their coagulation laboratory practice and EQA. RESULTS: From a dispatch of 220 questionnaires, 124 (56.4%) were returned. Of the 112 hospitals that had coagulation tests, all of them performed prothrombin time (PT), and 110 laboratories performed activated partial thromboplastin time (APTT) as well. Thirty eight percent of laboratories still used 3.8% sodium citrate as the anticoagulant for coagulation tests. The majority of laboratories (65%) reported normal control value with the patient results. Only 42% of coagulation laboratories established their own reference range. The denominators of PT ratio and APTT ratio calculations were derived from several sources apart from the mean of normal subjects. Seven of 112 (6%) laboratories participated in an EQA program. CONCLUSION: The present survey represents an overview of the current laboratory practice for coagulation tests in Thailand Improvement is necessary, and the survey results emphasize the need for establishing an EQA program in Thailand PMID- 18386712 TI - Comparable between rapid one step immunochromatographic assay and ELISA in the detection of prostate specific antigen in vaginal specimens of raped women. AB - BACKGROUND: Rape is a crime found in Thailand nowadays. The crime is often lacking of eyewitnesses. Therefore, examination for forensic biological evidence becomes quite important, especially investigating sperm and semen in vaginal specimens of the victim. Acid phosphatase test for semen is commonly used in Thailand but is just a presumptive test. Recently, confirmatory kit tests became available in Thailand for detecting the prostate specific antigen (PSA) from semen. This test is simpler and cheaper than ELISA. OBJECTIVE: To compare the rapid one-step immunochromatographic assay with ELISA for the detection of prostate specific antigen in vaginal specimens of raped women. MATERIAL AND METHOD: A diagnostic test was conducted on the vaginal specimens of raped women that were sent to the laboratory of the Pathology Department, Faculty of Medicine, Ramathibodi Hospital, Mahidol University during April-August 2006. One hundred vaginal specimens were examined for prostate specific antigen by rapid one step immunochromatographic assay and compared with ELISA. RESULTS: There were 85% and 83% of sensitivity, 85% and 85% of specificity, 85% and 85% of accuracy, 89% and 89% of positive predictive value, and 79% and 77% of negative predictive value from rapid one-step test kit and ELISA respectively CONCLUSION: The result showed that there was no difference on specificity, accuracy and positive predictive value between the two methods but sensitivity and negative predictive value of rapid one-step test kit was better than ELISA. The research team recommends that rapid one-step test kit for prostate specific antigen should be routine service in vaginal specimens of raped women. PMID- 18386713 TI - Invasive fungal infection in Ramathibodi Hospital: a ten-year autopsy review. AB - OBJECTIVE: Determine the clinicopathological findings in autopsy cases with invasive fungal infection. MATERIAL AND METHOD: The autopsy and medical records with invasive fungal infection in Ramathibodi Hospital between January 1997 and December 2006 were analyzed. The criterions for the diagnosis of invasive fungal infection were the evidence of fungal elements from histopathological section. The age, gender underlying predisposing risk factors for the disease, clinical manifestations, extent of systemic organ involvement documented morphologically at autopsy, and fungal culture were analyzed RESULTS: There were 155 autopsy cases (73 male, 82 female; mean age 45.3 years, range 3 months to 87 years) with the diagnosis of invasive fungal infection. The common clinical presentations were fever (55.5%), and dyspnea (26.5%). The invasive fungal infection was associated with hematologic malignancy in 31%. The common mycoses were aspergillosis and candidiasis, which were observed in 88 and 80 cases, respectively. There were 32 cases (20.6%) of mixed fungal infection. Cultures from autopsy materials were positive for fungus in 80 cases out of 99 cases (80.8%). The most frequent site of fungal infection was in the lungs (74.8%), followed by gastrointestinal tract (28.4%), and brain (26.5%). Invasive fungal infection was diagnosed intravitally in 63.9% of total cases. CONCLUSION: A diagnosis of invasive fungal infection requires a high index of suspicion, especially in immunocompromised patients who presented with prolonged fever Clinical specimens must be sent for histopathology and fungal culture for a definite diagnosis and an appropriate management. Therefore, the physician should inform the laboratory if invasive fungal infection is suspected because special media are necessary for the best recovery of fungi. In addition, the present study underscores the significance of autopsy as a diagnostic method and means of medical quality control. PMID- 18386714 TI - A laparoscopic versus open adrenalectomy in Ramathibodi Hospital. AB - OBJECTIVE: To compare the results of laparoscopic adrenalectomy with those of open adrenalectomy in Ramathibodi Hospital. MATERIAL AND METHOD: Medical charts of 41 laparoscopic and 39 open adrenalectomy patients were reviewed Baseline characteristics and outcomes of treatment were compared between these two patient groups, using univariable statistical tests and multivariable statistical procedures. RESULTS: There were significant baseline differences between the two groups in terms of gender, body mass index, ASA class, and preoperative diagnosis. The outcomes operative time, estimated blood loss and length of hospital stay were also significantly different. After adjusting for the effects of baseline differences, laparoscopic adrenalectomy was associated with a significant reduction of length of hospital stay by 40%. CONCLUSION: Laparoscopic adrenalectomy is a safe and effective procedure and should help hasten postoperative recovery and may save the costs of hospitalization. PMID- 18386715 TI - Laparoscopic radical prostatectomy: transperitoneal laparoscopic radical prostatectomy versus extraperitoneal endoscopic radical prostatectomy. AB - OBJECTIVE: To compare the perioperative results between Transperitoneal Laparoscopic Radical Prostatectomy (T-LRP) and Extraperitoneal Endoscopic Radical Prostatectomy (E-LRP). MATERIAL AND METHOD: Retrospective reviews of 125 patients who underwent laparoscopic radical prostatectomy by single surgeon (C.N) for stage T2-T3 adenocarcinoma of the prostate between May 2001 and July 2006 at Siriraj Hospital. Fifty-six cases had T-LRP and 69 cases had E-LRP The preoperative data (age, presenting PSA, and Gleason score), perioperative data (prostatic weight, operative time, intraoperative blood loss, the day of full oral diet, length of drain, and catheter time), pathologic stage, and margin status were compared. RESULTS: Mean age and Gleason score were comparable in both groups. Mean presenting PSA was lower in T-LRP (9.93) as compared to E-LRP (21.84) (p = 0.046). The mean prostatic weight was comparable in both T-LRP and E LRP. The mean operative time of T-LRP (350) was significant longer than E-LRP (220) (p < 0.001). Mean intraoperative blood loss was more in T-LRP (883) as compared to E-LRP (605) (p = 0.001). Average blood transfusion was higher in T LRP (1.23 unit) as compared to E-LRP (0.32). Postoperative full oral diet, length of drain, and catheter time in E-LRP were shorter than T-LRP (full diet: median 2 days vs. 3 days, p = 0.001) (length of drain: 4.98 days vs. 6.69 days, p = 0.002) (Catheter time: 8.9 days vs. 11.9 days, p = 0.002). Margin status were comparable in both groups but mean postoperative Gleason score was higher in E-LRP as compared to T-LRP (7.2 vs. 6.85, p = 0.022). CONCLUSIONS: E-LRP resulted in significant less operative time, intraoperative blood loss, postoperative oral diet, length of drain and catheter time where as the pathological margin status was the same in both T-LRP and E-LRP. PMID- 18386716 TI - Causes of corneal blindness: a multi-center retrospective review. AB - OBJECTIVE: To assess prevalence and causes of corneal blindness in four hospitals in Thailand and to compare the causes between hospitals. MATERIAL AND METHOD: A retrospective chart review of six hundred and sixty four patients records with best-corrected visual acuity not better than 6/60 in at least one eye due to corneal disease in Siriraj, Mettapracharak, Sappasitthiprasong and Banmee Hospitals in a one-year period were reviewed. The collected data included age, sex, laterality, causes of corneal blindness, and prognosis. RESULTS: Seven hundred and sixty nine eyes with corneal blindness and a median age of 55.0 years were reviewed Leading causes of cornea blindness were corneal infection (35.6%), surgical bullous keratopathy (27.8%), and trauma (14.0%). The prevalence at Siriraj Hospital was 1.7% (498/28,728 patients). Most of the treatable cases (84.6%) underwent penetrating keratoplasty procedures and 73% of patients had a chance of recovery. CONCLUSION: Corneal infection and surgical bullous keratopathy were the leading causes of corneal blindness. PMID- 18386717 TI - Evaluation of the Thai, Low Vision Quality-of-Life Questionnaire (LVQOL). AB - BACKGROUND: Quality of life is an important measurement of medical outcome. The assessment questionnaire for low vision patients allows the services to be analysed, developed and improved. OBJECTIVE: To evaluate the reliability and validity of the Thai version of the Low Vision Quality-of-Life Questionnaire (LVQOL). MATERIAL AND METHOD: A cross sectional study was performed, and the content validity was examined by factor analysis and reliability assessed by the Cronbach's alpha coefficient. 47 respondents participants were interviewed at the Department of Ophthalmology, Mettapracharak Eye Center. RESULTS: The Thai version of the LVQOL achieved good levels of validity and reliability which the Cronbach's alpha coefficient equaled to 0.7224-0.9099 among 22 items. CONCLUSION: The Thai version of the LVQOL can be used as an instrument to evaluate the low vision quality of life. It is useful in determining the effects of the low vision rehabilitation. PMID- 18386718 TI - Blast injury of the ears: the experience from Yala Hospital, Southern Thailand. AB - OBJECTIVE: To report acute and subacute consequences of blast injury to the ear from terrorists' bombings experienced from Yala Provincial Hospital, Southern Thailand STUDY DESIGN: Retrospective chart review was done on 54 patients who suffered otologic injuries from bombing attacks in Yala Province from January to May 2005. Only 33 patients who had complete otologic and audiologic examination with a 3-month follow-up were studied. MATERIAL AND METHOD: The ear symptoms, the size of tympanic membrane perforation, degree of hearing loss at first examination within 30 days after injuries were recorded. After a 3-month follow up, the rate of spontaneous healing, rate of operation needed and long-term complications including hearing loss were analyzed. RESULTS: The two most common initial symptoms were hearing loss (72.73%) and tinnitus (66.67%). Tympanic membrane perforations were encountered in 31 ears of 22 out of 33 patients. Spontaneous healing occurred in 23 ears (74.19%) with the highest incidence in small perforations (size < 50%). All healings occurred within 8 weeks. Tympanoplasty was done on the rest, except one patient. Eight patients (24.24%) had sensorineural hearing loss without tympanic membrane perforation. They still have sensorineural hearing loss, which is rather mild and typically affects in high tone with five of this group having normal hearing in speech range. Eleven patients from the tympanic membrane perforation group still have mixed hearing loss, which were also mostly mild. CONCLUSION: Patients with aural symptoms after a blast injury need thorough otologic and audiological examination. The spontaneous healing of tympanic membrane perforation from explosive injury was relatively high (74.19%) after an 8-week follow-up, only 8/31 ears required surgical repair. At 3-months follow-up, more than two-thirds of the patients still had residual hearing loss, which was rather mild and affected mainly in high tone. PMID- 18386719 TI - Local injection of dexamethasone for the treatment of carpal tunnel syndrome in pregnancy. AB - OBJECTIVE: To evaluate the results of 4 mg of dexamethasone acetate injections for the treatment of carpal tunnel syndrome in pregnancy. MATERIAL AND METHOD: Twenty-four pregnant volunteers who suffered from carpal tunnel syndrome occurring in the third trimester of their pregnancies were injected with 4 mg of dexamethasone acetate combined with 1% lidocaine into the carpal tunnels. The volunteers filled the Boston symptom severity questionnaire and were examined by Phalen's test, Semmes-Weinstein monofilament test, grip strength, and pinch strength before injection, one month after injection, and one month after delivery. RESULTS: There was significant improvement of symptoms, grip strength and pinch strength of the injected hand compared with the opposite side at one month after injection (p < 0.01). However, after delivery, the symptoms of carpal tunnel syndrome improved on both the injected and opposite hands. CONCLUSION: Carpal tunnel syndrome in pregnancy is generally resolved after delivery and, therefore, should be treated conservatively. The patient with severe symptoms can be treated with dexamethasone injection in the third trimester with good results. PMID- 18386720 TI - Renal microvascular changes in streptozotocin-induced, long-termed diabetic rat. AB - OBJECTIVE: To investigate the renal microvascular changes in streptozotocin (STZ) induced, long-termed diabetic rat. MATERIAL AND METHOD: Twelve male Sprague Dawley rats were used. Each diabetic rat (n = 8) was induced by an intraperitoneal injection of STZ (60 mg/kg) in citrate buffer (pH 4.5). Control rats (n = 4) were injected intraperitoneally with the same amount of the buffer. The animals were sacrificed at 20 weeks after the injections. The kidneys were processed for conventional light microscopy (LM) and vascular corrosion cast technique with scanning electron microscopy (SEM). RESULTS: Under LM, it was found that the glomerular sizes intensively decreased in the long-termed diabetic rat. The thickening of Bowman's basement membrane was demonstrated. Additionally, there were macrophages and capsular drop lesions in renal corpuscles of long termed diabetes. The sizes of proximal and distal tubules were markedly destroyed, when compared to the control. Moreover, the epithelial necrosis of vacuolated renal tubules was observed. By using vascular corrosion cast with SEM, the glomerular microvascular sizes in the long-termed diabetes were significantly decreased that corresponded to the result under LM. Furthermore, the size of peritubular capillaries decreased. Concerning to vasa recta in the long-termed diabetes, these vessels ran tortuously and decreased in size. CONCLUSION: Renal microvascular changes, observed in STZ-induced diabetic rats, mimic human diabetic nephropathy (DN). Additionally, the pathological changes of the renal tubules were investigated. Therefore, the present study provides an important basic knowledge for understanding the processes in developing DN, as well as for further study of the therapeutic treatment. PMID- 18386722 TI - Abnormal diffusivity of normal appearing brain tissue in multiple sclerosis: a diffusion-weighted MR imaging study. AB - OBJECTIVE: To assess whether water diffusivity in normal appearing brain tissue including white and gray matter of multiple sclerosis (MS) patients shown by diffusion-weighted imaging (DWI) differs from normal individuals. MATERIAL AND METHOD: Conventional MRI and DWI were performed in 37 multiple sclerosis patients and 31 control subjects, matched for age and sex. Quantitative diffusivity values were obtained from variable locations of normal appearing white and gray matter from both hemispheres by using a standardized region of interest template. +/- 2. 9 x 10(-5) mm2/s and 85.90 x 10(-5) +/- 2.45 x 10(-5) mm2/s) than normal control subjects (NAWM: 73.46 x 10(-5) +/- 1.77 x 10(-5) mm2/s and NAGM: 82.90 x 10(-5) +/- 0.91 x 10(-5) mm2/s) with p-value < 0.0001. CONCLUSION: Water diffusivity was higher in all NAWM regions, deep gray matter regions, and some cortical gray matter region of MS patients than normal controls. DWI can quantify the presence and extent of MRI-undetectable pathology in the normal appearing brain tissue that were the disease burden. PMID- 18386721 TI - A comparison of propofol-LCT with propofol-LCT/MCT on pain of injection. AB - BACKGROUND: Propofol-Lipuro 1% is consisting of long-chain triglycerides (LCT) and medium-chain triglycerides (MCT) that have been reported to reduce injection pain. OBJECTIVE: To compare the incidence and intensity of injection pain with 1% Propofol-LCT with 1% Propofol-LCT/MCT in female populations for minor surgery under total intravenous anesthesia (TIVA). MATERIAL AND METHOD: One hundred and ten female patients were double-blind randomized into two groups. Group long chain triglycerdes (L) received 1% propofol-LCT while group long-chain triglycerides/medium-chain triglycerides (L/M) received 1% propofol-LCT/MCT. All patients received no benzodiazepine premedication and fentanyl 1 microg/kg was given 3 minutes before propofol injection. The propofol 1 mg/kg was manually injected at 0.5 mL/sec. The verbal rating score (VRS 0-10) was recorded on pain of injection. VRS > 4 indicates a significant response to pain. RESULTS: There was a significantly greater incidence and intensity of injection pain in group L compared with group L/M (p < 0.001 and p = 0.013 respectively). CONCLUSIONS: Propofol-LCT/MCT is superior to propofol-LCT on reducing pain of injection. PMID- 18386723 TI - Flexor tendon rupture after distal radius fracture. Report of 2 cases. AB - Flexor tendon rupture after distal radius fracture is very rare. The authors reported two cases. The first case had flexor pollicis longus tendon rupture. The other had flexor pollicis longus and multiple finger flexor tendons rupture. The authors propose two modes of mechanism of tendon rupture after distal radius fracture. PMID- 18386724 TI - Bisphosphonate-related osteonecrosis of the jaws: a call for multidisciplinary approaches. AB - Bisphosphonates have been prescribed for the treatments of oncologic and metabolic bone diseases to inhibit bone resorption of osteoclasts. However, in recent years, the increased numbers of cases diagnosed with exposed and necrotic bone localized in the jawbones associated with bisphosphonate use have been reported, mostly in patients with multiple myeloma or bone metastases who received long-term intravenous bisphosphonate treatments. The strong association between patients receiving dentoalveolar surgery and the incidence of this complication highlights the need for multidisciplinary approaches and necessitates the close attention from a team of health care personnel. The present review summarizes the current knowledge on etiology, risk factors, clinical presentations, and recommended preventive measures and managements for afflicted patients. In light of recent available data and because stanterdized management strategies have not been well established, prevention seems to be of paramount benefit to this group of patients. PMID- 18386725 TI - Enhanced recovery programmes in colorectal surgery: clinical implications. PMID- 18386726 TI - Maternal safety: what is the cost? PMID- 18386728 TI - Lessons learnt from the reorganization of maternity services. AB - Recent experiences of working with maternity services in difficulty have highlighted the need for regular surveillance of clinical governance parameters, using a performance chart or 'dashboard' to monitor and assure quality and safety. PMID- 18386729 TI - Obstetric services in the UK: what we need now. AB - Pregnancy and childbirth should be a safe and memorable experience for both women and their partners. This article discusses the current position of maternity services, their strengths and limitations, and suggests future directions required to improve these services further. PMID- 18386730 TI - Obstetrics: what we need in the future. AB - Health-care professionals who work in obstetrics have seen many changes over the last decades, with both good and bad results. But what changes could be made to improve the future care of pregnant women? PMID- 18386731 TI - The changing face of coeliac disease. AB - Coeliac disease was considered to be a rare childhood affliction causing malabsorption and weight loss. It is now recognized to occur at any age with a range of non-specific manifestations including anaemia, recurrent miscarriages, neuropsychiatric disorders and osteopenia. This article summarizes advances in diagnosis and innovations in therapy. PMID- 18386732 TI - Antidepressants: will new mechanisms of action improve poor outcomes? AB - Depression is one of the most common mental illnesses and has a high impact on individuals and society. Despite the numerous treatment approaches, poor response combined with the burden of residual symptoms negatively affects the overall outcome. Novel pharmacological approaches could help to address this issue. PMID- 18386733 TI - Ultrasound imaging of the scrotum. PMID- 18386734 TI - Practical management of inpatient diabetes regimens. PMID- 18386735 TI - Safeguarding your health: a guide for the foundation year doctor. PMID- 18386736 TI - Voluntary work overseas for foundation doctors. PMID- 18386737 TI - Writing a book review: frequently asked questions answered. PMID- 18386738 TI - Medical conditions in works of art. AB - With their stark contrast and blurred boundaries, the relationship between science, medicine and art has long been a fascinating area of exploration. The depiction of medical conditions in art has enhanced our understanding of the evolution of disease and its treatment, but exact diagnosis of the underlying medical conditions can be difficult. This article reviews some paintings suggesting underlying medical diagnoses under specialty-based headings. PMID- 18386739 TI - Surgical management of morbid obesity. AB - Obesity affects 1.7 million people worldwide, and is a leading cause of morbidity and mortality. Outcomes of conservative management are generally poor. Surgery for morbid obesity has reported excellent results, leading to an exponential increase in numbers of procedures performed. PMID- 18386740 TI - Analysis of selection tools for appointment of specialty trainees in histopathology in Scotland. AB - This study explores possible benefits of structured selection methods in the employment of specialty trainees in histopathology in Scotland. It examines methods used in 2007 demonstrating novel approaches and discusses the positive reaction of consultants involved in the process. PMID- 18386741 TI - Spontaneous splenic rupture in the postpartum period. PMID- 18386742 TI - An unusual cause of nausea and altered bowel habit. PMID- 18386743 TI - Neurosarcoidosis presenting as ghost lesions in the CNS: a diagnostic dilemma. PMID- 18386744 TI - Scleral pigmentation and a heart murmur. PMID- 18386745 TI - Coarctation of the aorta, hypertension and associated features. PMID- 18386746 TI - The case for individual savings accounts. PMID- 18386747 TI - John Snow: early anaesthetist and pioneer of public health. PMID- 18386748 TI - Anaesthetic rooms: luxury or necessity? PMID- 18386749 TI - Radiology practice management in Canada: changing times, increased need. PMID- 18386750 TI - Strategic planning in diagnostic imaging: meeting the challenge. AB - PURPOSE: The 21st century has raised new and significant challenges to the practice of diagnostic imaging. Radiologists will have to navigate a difficult path as they face threats from disruptive technologies, adverse demographic changes, pressures to limit reimbursement both from the public and from the private sectors, increased domestic and crossborder competition, and many others. Success in these trying times will require greater attention to strategic planning if we are to thrive and survive in radiology. METHODS: Strategic planning and tactical implementation methodologies were reviewed, from Sun Tzu to the present day, for applicability to the needs of modern radiology groups. RESULTS: A framework for developing and implementing strategic plans was constructed to assist radiology leaders and groups in considering the spectrum of tasks, from gathering intelligence, to developing scenarios, to implementing and evaluating tactical plans. CONCLUSIONS: Strategy and tactics are too important to be ignored or left to others. They need to be core activities for all radiology leaders. Frameworks can be used to help in providing structure and rigour to strategic planning efforts at the department and group level. PMID- 18386751 TI - The fundamentals of negotiation. PMID- 18386752 TI - Medical invention: transforming your idea into a product. PMID- 18386753 TI - Can ultrasound evaluate axillary lymph node status in patients with breast cancer? PMID- 18386754 TI - Performance of carotid stenting, vertebroplasty, and EVAR: how many are we doing and why are we not doing more? A survey by the Canadian Interventional Radiology Association. AB - OBJECTIVE: To determine the percentage of interventional radiologists who currently perform 3 interventional procedures: carotid stenting, vertebroplasty, and endovascular aneurysm repair (EVAR) in Canada, and impediments to their future performance by other interventional radiologists. METHODS: An anonymous online survey was emailed to all members of the Canadian Interventional Radiology Association (CIRA). The survey was open for a period of 2 months. RESULTS: A total of 75 survey responses were received (of an estimated 247). Carotid stenting, vertebroplasty, and EVAR were performed at 40%, 59%, and 46% of respondents' centres respectively. Wait times, from referral to consultation, and from consultation to procedure, were both typically between 2 to 4 weeks, longer for EVAR. Of respondents currently not performing these procedures, 26%, 28%, and 16% anticipated beginning to perform carotid stenting, vertebroplasty, and EVAR, respectively, in the proceeding year from time of survey. Of respondents who wished to perform the procedure, the greatest impediments were a lack of training, lack of a referral base, and lack of support from their radiology department and (or) colleagues. CONCLUSIONS: Although carotid stenting, vertebroplasty, and EVAR were being performed at about one-half of respondent's centres, and there will likely be greater adoption of the procedures in the near future, there remain substantial impediments. The greatest impediments to additional radiologists performing these procedures were a lack of training, lack of referral base, and lack of support from their radiology department and (or) colleagues. The former impediment suggested an unmet need for additional training courses. PMID- 18386755 TI - Answer to case of the month #127. Bilateral internal carotid artery dissections with pseudoaneurysm. PMID- 18386756 TI - Answer to case of the month #128. Spectral waveform owing to intra-aortic balloon pump counter pulsation. AB - The intra-aortic balloon pump (IABP) is inserted percutaneously, usually through the femoral artery with placement in the descending aorta with its tip at the distal aortic arch (below the origin of the left subclavian artery). The balloon is connected to a drive console, consisting of a pressurized gas reservoir, a monitor for ECG and pressure wave recording, adjustments for inflation and deflation timing, triggering selection switches and battery backup power sources. The gases used for inflation are either helium or carbon dioxide. The advantage of helium is its lower density and therefore a better rapid diffusion coefficient. Carbon dioxide has increased solubility in blood and thereby reduces the potential consequences of gas embolization following a balloon rupture. PMID- 18386757 TI - Answer to case of the month #129. Mueller-Weiss syndrome. PMID- 18386758 TI - The bruised face in a hematoma. PMID- 18386759 TI - Paediatric kidney transplantation in Belgium. AB - In Belgium, kidney transplantation is currently the treatment of choice for a child with end-stage renal disease (ESRD). Dialysis remains the life-saving bridge to transplantation. Within the Eurotransplant (ET) community, Belgium represents 14% of the cadaveric transplantations and 22% of the living-related transplantation (LD) in children less than 16 years of age. Single-centre analysis (KUL) shows a patient survival of 94% at 3 year and 91% at 5 year. The overall graft survival is 82% at 3 year and 74% at 5 year. In the LD group, the graft survival rate is 10% better than the overall actuarial graft survival rate. Multivariate Cox regression analysis performed on all transplantations of one centre (KUL) demonstrate the following factors to be significant and independent predictors of poor graft outcome: absence of calcineurin inhibitors, two HLA- mismatches, duration of pre-transplant dialysis and creatinine clearance at one year after transplantation. The outcome improves by a short dialysis waiting time, the use of living-related donors, the prevention of delayed graft function (DGF), and of acute rejection. Within the ET community, the waiting child has priority compared to the adult, but if we want to avoid morbidity, waiting times must be shortened and the incidence of pre-emptive transplantation, which is currently 24% in Belgium, must increase. The good results with LD is certainly an attractive alternative to be actively encouraged for paediatric kidney recipients and the use of young deceased donors especially for children with ESRD must be supported since the results in terms of graft survival with these donors are very good, especially in children. In paediatric kidney transplantation the long-term graft survival is still the major challenge and has still to be documented by randomized trials. The success of the past, however, allows us to face the future with hope and confidence. PMID- 18386760 TI - Kidney in old age. AB - Epidemiological studies have shown that chronic kidney disease (CKD) is a major health problem especially in an older population. Given the growing segment of the elderly population and the important implication of renal disease on health care, this review discusses the structural and functional changes of the ageing kidney and the underlying mechanisms of age-dependent injury. The implications of these changes in daily clinical practice and the management of CKD is also briefly overviewed. PMID- 18386761 TI - Developmental pharmacology: neonates are not just small adults... AB - Neonatal drug dosing needs to be based on the physiological characteristics of the newborn and the pharmacokinetic parameters of the drug. Size-related changes can in part be modelled based on allometry and relates to the observation that metabolic rate relates to weight by a kg 0.75 trend. Until adult metabolic activity has been reached, ontogeny, i.e. isoenzyme-specific maturation and maturation of renal clearance also contributes to drug metabolism, making isoenzyme-specific documentation of maturation necessary. Changes in body composition and ontogeny are most prominent in neonates. The body fat content (/kg) is markedly lower and the body water content (/kg) is markedly higher in neonates. These findings have an impact on the distribution volume of both lipophilic and hydrophilic drugs. Drugs are cleared either by metabolism or elimination. While the first is mainly hepatic, the second route is mainly renal. Both hepatic metabolism and renal clearance display maturation in early life although other covariables (e.g. polymorphisms, co-administration of drugs, first pass metabolism, disease characteristics) further contribute to the interindividual variability in drug disposition. Documentation of these maturational processes based on in vivo 'case' studies is of value since these drug-specific observations can subsequently be extrapolated to other drugs which are either already being prescribed or even considered for use in neonates by the introduction of these observations in 'generic physiologically-based pharmacokinetic' models. PMID- 18386762 TI - A registry of patients treated with drotrecogin alfa (activated) in Belgian intensive care units--an observational study. AB - BACKGROUND: Drotrecogin alfa (activated) [DrotAA] is the only specific sepsis therapy that has been shown to reduce mortality. The objectives of this study were to document the profile of patients treated with DrotAA in Belgian intensive care units (ICUs), using data from a database established as part of drug reimbursement conditions in Belgium, and to compare the observed hospital mortality of these patients with their expected mortality, calculated using data from non-DrotAA-treated patients from the Belgian section of PROGRESS, a separate, voluntary, international sepsis registry collecting data from patients with severe sepsis. MATERIAL AND METHODS: Data from the non-DrotAA-treated patients in PROGRESS were used to calculate the expected mortality rates for DrotAA-treated patients in the Belgian registry. Using a logistic regression equation, these rates were controlled for age and the presence or absence of organ dysfunction in each of 5 organ systems. The same logistic regression technique was used to control the mortality rates observed in the DrotAA-treated patients from the Belgian registry for age and the presence or absence of each of the 5 organ dysfunctions. Adjusted expected and observed hospital mortality rates could then be compared. RESULTS: There were 436 DrotAA patients in the Belgian registry. Almost all the patients (99.5%) had at least 2 organ failures and the hospital mortality was 51.6%. Two hundred and eighty-six of the patients had enough baseline data to be included in the regression model. Using data from the PROGRESS non-DrotAA patients, the predicted hospital mortality, controlled for age and organ dysfunction, of Belgian registry patients, had they not been treated with DrotAA, was 63.5%. The observed hospital mortality, again controlled for age and organ dysfunction, of the 286 Belgian registry patients was 50.7%, implying an adjusted absolute mortality reduction of 12.8%. CONCLUSIONS: Comparing Belgian reimbursement registry data with those of a voluntary severe sepsis register provides support for the observation that DrotAA reduces mortality rates in severe sepsis and septic shock. PMID- 18386763 TI - Costs and length of stay associated with antimicrobial resistance in acute kidney injury patients with bloodstream infection. AB - INTRODUCTION: Antimicrobial resistance negatively impacts on prognosis. Intensive care unit (ICU) patients, and particularly those with acute kidney injury (AKI), are at high risk for developing nosocomial bloodstream infections (BSI) due to multi-drug-resistant strains. Economic implications in terms of costs and length of stay (LOS) attributable to antimicrobial resistance are underevaluated. This study aimed to assess whether microbial susceptibility patterns affect costs and LOS in a well-defined cohort of ICU patients with AKI undergoing renal replacement therapy (RRT) who developed nosocomial BSI. METHODS: Historical study (1995-2004) enrolling all adult RRT-dependent ICU patients with AKI and nosocomial BSI. Costs were considered as invoiced in the Belgian reimbursement system, and LOS was used as a surrogate marker for hospital resource allocation. RESULTS: Of the 1330 patients with AKI undergoing RRT, 92 had microbiologic evidence of nosocomial BSI (57/92, 62% due to a multi-drug-resistant microorganism). Main patient characteristics were equal in both groups. As compared to patients with antimicro-4 bial-susceptible BSI, patients with antimicrobial-resistant BSI were more likely to acquire Gram-positive infection (72.6% vs 25.5%, P<0.001). No differences were found neither in LOS (ICU before BSI, ICU, hospital before BSI, hospital, hospital after BSI, and time on RRT; all P>0.05) or hospital costs (all P>0.05) when comparing patients with antimicrobial resistant vs antimicrobial-susceptible BSI. However, although not statistically significant, patients with BSI caused by resistant Gram-negative-, Candida-, or anaerobic bacteria incurred substantial higher costs than those without. CONCLUSION: In a cohort of ICU patients with AKI and nosocomial BSI undergoing RRT, patients with antimicrobial-resistant vs antimicrobial-susceptible Gram positive BSI did not have longer hospital stays, or higher hospital costs. Patients with resistant "other" (i.e. Gram-negative, Candida, or anaerobic) BSI were found to have a distinct trend towards increased resources use as compared to patients with susceptible "other" BSI, respectively. PMID- 18386764 TI - Pulsating tumour in the clavicular region of a chronic haemodialysis patient with ipsilateral vascular access catheter. AB - We describe the case of a 70-year-old woman who presented with a pulsating mass in the left supraclavicular region during a haemodialysis session. The frequency of the pulsations, parallel to that of the dialysis-related blood flow cycle, without blood loss at the exit site, indicated that this observation could potentially be attributed to a rupture of the silastic material in the subcutaneous track of the catheter. Our hypothesis was confirmed after removal of the catheter. PMID- 18386765 TI - Safety of alternative medicines reconsidered: lead-induced anaemia caused by an indian ayurvedic formulation. AB - We report two cases of Belgian women diagnose with a lead poisoning. Both patients presented with abdominal pain and a normochromic normocytic anaemia. The diagnosis was based on the clinical symptoms, the anaemia, the basophilic stippling of erythrocytes and the elevated blood lead level Upon further questioning, both patients reported the use of Ayurvedic medications. Toxicological analysis of the different pills revealed that, in both, the same orange-red pills contained a remarkably high amount of lead. Cases of lead poisoning associated with the use ofAyurvedic formulations are emerging around the world. However, to our knowledge, these are the first reported cases in Belgium. PMID- 18386766 TI - Exploratory assessment of dose proportionality: review of current approaches and proposal for a practical criterion. AB - This article reviews currently used approaches for establishing dose proportionality in Phase I dose escalation studies. A review of relevant literature between 2002 and 2006 found that the power model was the preferred choice for assessing dose proportionality in about one-third of the articles. This article promotes the use of the power model and a conceptually appealing extension, i.e. a criterion based on comparing the 90% confidence interval for the ratio of predicted mean values from the extremes of the dose range (R(dnm)) to pre-defined equivalence criterion (theta(L),theta(U)). The choice of bioequivalence default values of theta(L)=0.8 and theta(U)=1.25 seems reasonable for dose levels only a doubling apart but are impractically strict when applied over the complete dose range. Power calculations are used to show that this prescribed criterion lacks power to conclude dose proportionality in typical Phase I dose-escalation studies. A more lenient criterion with values theta(L)=0.5 and theta(U)=2 is proposed for exploratory dose proportionality assessments across the complete dose range. PMID- 18386767 TI - Molecularly targeted therapy and cancer surgery. PMID- 18386768 TI - Caveats in the interpretation of the surgical literature. AB - The ability to appraise research literature critically is an invaluable tool in the era of evidence-based medicine. The objective of this educational article is to highlight certain caveats that are important to those who seek to interpret the surgical literature. These caveats are illustrated with intuitive examples drawn from the field of surgery. It is hoped that the article will help surgeons to recognize the common pitfalls encountered when interpreting scientific reports. This is important if research findings are to be transferred appropriately into the realm of clinical practice. PMID- 18386769 TI - Multivariable analysis comparing outcome after sentinel node biopsy or therapeutic lymph node dissection in patients with melanoma (Br J Surg 2007; 94: 1293-1299). PMID- 18386770 TI - Role of transplantation in the management of hepatic malignancy (Br J Surg 2007; 94: 1319-1330). PMID- 18386772 TI - Impact of anastomotic leakage on oncological outcome after rectal cancer resection (Br J Surg 2007; 94 1548-1554). PMID- 18386774 TI - Frequent BRG1/SMARCA4-inactivating mutations in human lung cancer cell lines. AB - Components of the SWI/SNF chromatin-remodeling complex, such as INI1, are inactivated in human cancer and, thus, act as tumor suppressors. Here we screened for mutations the entire coding sequence of BRG1 (SMARCA4), which encodes the ATPase of the complex, in 59 lung cancer cell lines of the most common histopathological types. Mutations were detected in 24% of the cancer cell lines, many of them in cells commonly used for lung cancer research. All mutations were homozygous and most predicted truncated proteins. The alterations were significantly more frequent in the non-small-cell lung cancer (NSCLC) type (13/37, 35%) as compared to the small-cell lung cancer (SCLC) type (1/19, 5%) (P<0.05; Fisher's Exact test) and BRG1 was the fourth most frequently altered gene in NSCLC cell lines. BRG1 mutations coexisted with mutations/deletions at KRAS, LKB1, NRAS, P16, and P53. However, alterations at BRG1 always occurred in the absence of MYC amplification, suggesting a common role in lung cancer development. In conclusion, our data strongly support that BRG1 is a bona fide tumor suppressor and a major factor in lung tumorigenesis. PMID- 18386775 TI - Meta-analysis of sentinel node biopsy in ductal carcinoma in situ of the breast. AB - BACKGROUND: The need for sentinel lymph node (SLN) biopsy in patients with a preoperative diagnosis of ductal carcinoma in situ (DCIS) is debated. Advocates recommend such biopsy based on a high incidence of SLN involvement in some series. Opponents discourage SLN biopsy based on a perceived low incidence of nodal involvement in this setting. These contradictory arguments are generally based on small studies. The present study is a meta-analysis of the reported data on the incidence of SLN metastasis in patients with DCIS. METHODS: A search of electronic databases identified studies reporting the frequency of SLN metastases in DCIS. The random-effects method was used to combine data. RESULTS: Twenty-two published series were included in the meta-analysis. The estimate for the incidence of SLN metastases in patients with a preoperative diagnosis of DCIS was 7.4 (95 per cent confidence interval (c.i.) 6.2 to 8.9) per cent compared with 3.7 (95 per cent c.i. 2.8 to 4.8) per cent in patients with a definitive (postoperative) diagnosis of DCIS alone. This was a significant difference with an odds ratio of 2.11 (95 per cent c.i. 1.15 to 2.93). CONCLUSION: Patients with a preoperative diagnosis of DCIS should be considered for SLN biopsy. PMID- 18386776 TI - Effect of statins on proteolytic activity in the wall of abdominal aortic aneurysms (Br J Surg 2008; 95: 333-337). PMID- 18386780 TI - Leishmaniasis and concurrent hemophagocytosis with or without transient perforin expression perturbation. PMID- 18386781 TI - Smoking and binge drinking among Canadian survivors of childhood and adolescent cancers: a comparative, population-based study. AB - BACKGROUND: Objectives of this study were to describe the prevalence of smoking and binge drinking among survivors of childhood and adolescent cancer compared to controls who had never had cancer, and to identify factors associated with these behaviors. PROCEDURE: Data came from a national, multi-centre, population-based study of survivors of childhood and adolescent cancer (n = 1,263) aged 16 to 37 years and an age and gender matched control group (n = 1,422). Logistic regression analyses were performed to determine the likelihood and predictors of current smoking and binge drinking. RESULTS: Survivors were less likely to be current smokers (OR(adj) = 0.65, 95% CI = 0.54-0.77) and binge drinkers (OR(adj) = 0.66, 95% CI = 0.55-0.78) than the controls. Still, a substantial proportion of survivors reported smoking (23%) and binge drinking (25%). Survivors' smoking and binge drinking did not vary substantially by the clinical factors assessed. Survivors who received therapy associated with cardiac and/or pulmonary toxicity were as likely to smoke as non-exposed survivors. For both the survivors and controls current smoking and binge drinking were associated with lower education and higher reported stress. Binge drinking was also associated with being male and life dissatisfaction in both groups. CONCLUSIONS: This study indicates a need to reduce smoking and binge drinking among survivors of childhood and adolescent cancer and to design interventions addressing the underlying reasons for adopting unhealthy behaviors despite their risk for late effects. We identified factors related to smoking and binge drinking among survivors: being male, lower educational attainment, life dissatisfaction and high stress, which should help inform intervention development. PMID- 18386782 TI - Foot structure and footwear prescription in diabetes mellitus. AB - Foot structure abnormalities such as foot deformity and limited joint mobility are common and well established components of the diabetic foot which are associated with increased levels of mechanical stress on the foot and the development of ulcers. Our understanding of foot structure abnormality in diabetes has improved recently, mainly through the findings from in vivo imaging studies. Several examples will be discussed in this review. A limited understanding, however, still exists about several aspects related to the assessment, etiology, and consequences of change in foot structure in diabetes. Knowledge on these matters is needed if we are to better deal with the implications of foot structure change in diabetes. Diabetic patients with neuropathy and foot deformity are commonly prescribed with custom footwear, in particular after ulcer healing. The goal of this footwear is to redistribute and reduce plantar foot pressures, and to prevent ulcer recurrence. However, the available evidence for the effectiveness of custom footwear in secondary ulcer prevention is not yet strong. This may be associated with several factors, including a lack of standardized or systematic approach (a set of guidelines) in footwear prescription and evaluation or with the significant variability that exists across patients in the offloading effect of different footwear interventions, which increases the difficulty of predicting what works for a given patient. Objective evaluation tools such as in-shoe plantar pressure analysis can be helpful in this regard in order to ensure efficacy of an intervention. This provides a more optimal footwear solution that may lower the risk for ulceration. PMID- 18386783 TI - Perceived and actual motor competence of children treated for cancer with vincristine containing chemotherapy. PMID- 18386784 TI - Retrospective evaluation of pain assessment and treatment for acute vasoocclusive episodes in children with sickle cell disease. AB - BACKGROUND: This study was conducted to assess the care of pediatric patients hospitalized for sickle cell disease-related vasoocclusive episodes (VOE). The aim of this research was to illustrate the course of pain scores and methods of therapeutic intervention during hospitalization. PROCEDURE: Retrospective medical chart reviews were conducted to collect pain assessment and management data about children hospitalized during a 2-year period at an urban children's hospital. T tests and Chi-square analyses were used to identify differences in demographic variables, pain scores and opiate utilization. RESULTS: There were 59 children with 134 hospitalizations for VOE in a 2-year period. 50.8% of the patients were male; the mean age was 11.5 +/- 4.9 years. The average length of hospitalization was 4.6 +/- 2.7 days (range 1-19 days). Older patients stayed in the hospital significantly longer than younger patients (P = 0.002). Pain scores remained in the moderate to severe range (> or =5 out of 10) for many days in the majority of patients. Results failed to reveal significant differences in pain scores and opiate utilization between patients who had short versus extended hospitalizations, and for those patients with frequent versus infrequent hospitalizations for pain. CONCLUSIONS: Despite opiate dosing within recommended guidelines, mean pain scores remain in the moderate to severe range for several days following hospitalization for VOE. Future research should explore the factors which influence pain scores, as well as improved pain assessment and management techniques. PMID- 18386785 TI - Outcome analysis of childhood low-grade astrocytomas. AB - BACKGROUND: We aimed to determine the long-term natural history of low-grade astrocytomas (LGA) in children, with respect to pathology, and to evaluate influence of treatment on survival. PATIENTS AND METHODS: A consecutive cohort of patients < or =21 years with surgically confirmed LGA from 1965 to 1996 was assembled. All available pathology specimens were reviewed, masked to original diagnosis, patient data, and neuroimaging. RESULTS: Two hundred seventy-eight children (160 males; mean age 9.1 years; tumor location: 77 cerebrum, 62 cerebellum, 51 hypothalamic, 30 thalamus, 9 ventricle, 40 brainstem, and 9 spine) were assessed. Among 246 specimens reviewed, diagnoses were 135 pilocytic astrocytoma (PA), 27 diffuse astrocytoma (DA), 75 unclassifiable well differentiated astrocytoma (NOS), and 9 subependymal giant cell astrocytoma. At 5 and 10 years from initial surgery, for all LGA overall survival (OS) was 87% and 83%, while progression-free survival (PFS) was 55% and 42%, respectively. Original pathology diagnoses did not predict PFS (P = 0.47), but reviewed diagnoses were significantly associated with PFS (P = 0.007). Reviewed diagnoses were highly associated with OS (P < 0.0001), with 5-year OS for PA 96%, DA 48%, and NOS 86%; these differences remained significant when stratified by location or extent of resection. Among patients with residual tumor after surgery, 5-year PFS was 48% with observation alone (n = 114), no different (P = 0.32) from that achieved with immediate irradiation (n = 86). CONCLUSION: LGA, particularly PA, have excellent long-term OS. While tumor location and resection extent affect outcome, pathologic diagnosis when carefully interpreted significantly influences long-term survival. Immediate postoperative irradiation does not confer an advantage in delaying first progression in children with residual PA. PMID- 18386786 TI - CD11c/EYFP transgene illuminates a discrete network of dendritic cells within the embryonic, neonatal, adult, and injured mouse brain. AB - The CD11c enhanced yellow fluorescent protein (EYFP) transgenic mouse was constructed to identify dendritic cells in the periphery (Lindquist et al. [2004] Nat. Immunol. 5:1243-1250). In this study, we used this mouse to characterize dendritic cells within the CNS. Our anatomic results showed discrete populations of EYFP(+) brain dendritic cells (EYFP(+) bDC) that colocalized with a small fraction of microglia immunoreactive for Mac-1, Iba-1, CD45, and F4/80 but not for NeuN, Dcx, NG2 proteoglycan, or GFAP. EYFP(+) bDC, isolated by fluorescent activated cell sorting (FACS), expressed mRNA for the Itgax (CD11c) gene, whereas FACS anlaysis of EYFP(+) bDC cultures revealed the presence of CD11c protein. Light microscopy studies revealed that EYFP(+) bDC were present in the embryonic CNS when the blood-brain barrier is formed and postnatally when brain cells are amenable to culturing. In adult male mice, EYFP(+) bDC distribution was prominent within regions of the CNS that 1) are subject to structural plasticity and neurogenesis, 2) receive sensory and humoral input from the external environment, and 3) lack a blood-brain barrier. Ultrastructural analysis of EYFP(+) bDC in adult neurogenic niches showed their proximity to developing neurons and a morphology characteristic of immune/microglia cells. Kainic acid-induced seizures revealed that EYFP(+) bDC responded to damage of the hippocampus and displayed morphologies similar to those described for seizure-activated EGFP(+) microglia in the hippocampus of cfms (CSF-1R) EGFP mice. Collectively, these findings suggest a new member of the dendritic cell family residing among the heterogeneous microglia population. PMID- 18386787 TI - FGF-8 is involved in bone metastasis of prostate cancer. AB - Prostate cancer is the most common malignancy of men in Western countries. Patients with advanced prostate cancer suffer from incurable bone metastases. Recent data indicate that interactions between prostate cancer cells, osteoblasts, osteoclasts and the bone matrix are essential in the formation of bone metastases. FGF-8 is widely overexpressed in prostate cancer. Recently, FGF 8 has been found to affect both osteoblast and osteoclast differentiation. The aim of this study was to examine the role of FGF-8 in bone metastasis of prostate cancer. Immunohistochemistry was used to analyse FGF-8 expression in clinical samples of prostate cancer bone metastases. The functional significance of FGF-8 in growth of bone metastasis and formation of bone lesions was verified by using intratibial inoculations of FGF-8 or mock transfected PC-3 prostate cancer cells in nude mice. Intratibial tumors and bone lesions were analysed with X-ray, micro CT and detailed histomorphometry using image analysis software and with immunostaining for osteocalcin and cathepsin K. Immunohistochemical analysis of tissue microarray of bone metastases of human prostate cancer showed that 76% of human bone metastasis samples (n = 25 from 11 patients) were positive for FGF-8. FGF-8 increased the growth of intratibial tumors and local formation of lytic and sclerotic lesions of bone. These results demonstrate that FGF-8 is expressed at a high frequency in bone metastases of human prostate cancer and that expression of FGF-8 in PC-3 prostate cancer cells increases their growth as intratibial tumors and modulates formation of bone lesions in an in vivo model of prostate cancer bone metastasis. PMID- 18386788 TI - Polymorphisms in MGMT and DNA repair genes and the risk of esophageal adenocarcinoma. AB - Rates of adenocarcinoma of the esophagus (EAC) and esophago-gastric junction (EGJAC) have increased rapidly in recent decades. The primary risk factors, gastro-esophageal acid reflux and smoking, are potentially genotoxic through the generation of N-nitroso compounds. The DNA repair protein O(6)-methylguanine-DNA methyltransferase (MGMT) is the major cellular defense against alkylating DNA damage. We compared patients with EAC (n = 263) or EGJAC (n = 303) with matched population controls (n = 1,337) for the frequency of 5 MGMT single nucleotide polymorphisms (SNPs) (rs12269324, rs12268840, L84F, I143V, K178R), as well as SNPs in DNA repair genes ERCC1 (N118N), XRCC1 (Q399R) and XPD (K751Q). Relative risks were estimated using multivariable logistic regression. Potential biological interaction was assessed through the synergy index S. Each MGMT SNP conferred increased risks of EAC but not EGJAC; strongest associations were found for the 2 variant MGMT alleles rs12268840 and I143V (p = 0.005 and p < 0.001, respectively). Homozygous carriers of MGMT rs12268840 with frequent acid reflux had significantly higher risks of EAC (OR 15.5, 95% CI 5.8-42) than expected under an additive model, consistent with biological interaction (S = 3.3, 95% CI 1.1-10). Modest, nonsignificant interactions with smoking were also observed. Homozygous variant ERCC1 genotype was associated with reduced risks of EAC (OR 0.6, 95% CI 0.4-1.1), while the homozygous variant XRCC1 genotype conferred higher risks of EGJAC (OR 1.6, 95% CI 1.1-2.4). No associations with EAC or EGJAC were observed with XPD (rs13181). In summary, MGMT SNPs are associated with increased risks of EAC. Exposure to acid reflux, and possibly smoking, confer markedly higher risks among homozygous variant genotype carriers. PMID- 18386789 TI - beta-Ionone suppresses mammary carcinogenesis, proliferative activity and induces apoptosis in the mammary gland of the Sprague-Dawley rat. AB - beta-Ionone demonstrates potent anticancer activity both in vitro and in vivo. We determined tumor incidence and the number of rats bearing tumors as well as cell proliferation and apoptosis in a rat mammary cancer model induced by 7, 12 dimethylbenz[a]anthracene (DMBA). Rats were fed an AIN-76A diet containing beta ionone (0, 9, 18 or 36 mmol/kg), starting 2 weeks before DMBA administration and continuing for 24 weeks. A dose-dependent inhibition of mammary carcinogenesis by dietary beta-ionone was observed. Corresponding tumor incidence values were 82.1, 53.3, 25.9 and 10.0% (p < 0.01 or 0.05). Time to tumor appearance increased and tumor multiplicity decreased with increasing dietary beta-ionone. Histopathological and immunohistochemical evaluations of tumors were performed on the 64, 31, 15 and 3 tumors, respectively, identified in rats from the respective groups of 30. The proportions of adenocarcinomas, adenomas and benign masses were equally distributed in the latter group. In proportions within the other groups, the proportions of adenocarcinomas and benign masses decreased and increased with increasing dietary beta-ionone. Proliferating cell nuclear antigen (PCNA), cyclin D1 and Bcl-2 expression decreased, and Bax expression and nuclear fragmentation increased with increasing dietary beta-ionone. These results demonstrate the potent capacity of dietary beta-ionone to suppress DMBA-initiated mammary cancer in rats. PMID- 18386790 TI - Curcumin induces cell-arrest and apoptosis in association with the inhibition of constitutively active NF-kappaB and STAT3 pathways in Hodgkin's lymphoma cells. AB - Although treatment of Hodgkin's lymphoma (HL) with a multi-drug approach has been very successful, its toxicity becomes evident after several years as secondary malignancies and cardiovascular disease. Therefore, the current goal in HL treatment is to find new therapies that specifically target the deregulated signaling cascades, such as NF-kappaB and STAT3, which cause Hodgkin and Reed Sternberg (H-RS) cell proliferation and resistance of apoptosis. Based on the above information, we investigated the capacity of curcumin to inhibit NF-kappaB and STAT3 in H-RS cells, characterizing the functional consequences. Curcumin is incorporated into H-RS cells and acts inhibiting both NF-kappaB and STAT3 activation, leading to a decreased expression of proteins involved in cell proliferation and apoptosis, e.g. Bcl-2, Bcl-xL, cFLIP, XIAP, c-IAP1, survivin, c myc and cyclin D1. Interestingly, curcumin caused cell cycle arrest in G2-M and a significant reduction (80-97%) in H-RS cell viability. Furthermore, curcumin triggered cell death by apoptosis, as evidenced by the activation of caspase-3 and caspase-9, changes in nuclear morphology and phosphatidylserine translocation. The above findings provide a mechanistic rationale for the potential use of curcumin as a therapeutic agent for patients with HL. PMID- 18386791 TI - Combination of Fasl and GM-CSF confers synergistic antitumor immunity in an in vivo model of the murine Lewis lung carcinoma. AB - Gene transfer of Fas ligand (FasL) to tumor cells has been demonstrated to inhibit tumor growth in vivo, and neutrophils are primarily responsible for this immunoprotection. The granulocyte-macrophage colony stimulating factor (GM-CSF) secreted by tumor vaccine can recruit dendritic cells (DCs) for efficient antigen presentation to T cells that generate the tumor-specific response. To investigate whether the combination of FasL and GM-CSF can efficiently suppress tumor growth, we have established Lewis lung carcinoma (LLC-1) cells that are transduced with GM-CSF (LLC/GM-CSF), FasL (LLC/FasL) or both genes (LLC/FasL/GM-CSF) to test their tumorigenic potential in vivo. Mice inoculated with LLC/GM-CSF display high survival rates along with reduction of tumor growth. In contrast, none of the mice injected with LLC/FasL or LLC/FasL/GM-CSF develop tumors. Specific memory immune response and delayed LLC-1 tumor growth are found in mice immunized with LLC-1/FasL or LLC-1/FasL/GM-CSF. Furthermore, therapeutic effects are observed only when LLC-1/FasL/GM-CSF tumor vaccine is employed to retard growth of preexisting LLC-1 tumors. Tumor growth is also completely suppressed in mice injected with a mixture of LLC-1 and LLC-1/FasL/GM-CSF. In addition, IL-12 production, cytotoxic T-cell activity and IgG against LLC-1 are manifested in mice injected with LLC/FasL/GM-CSF. Our data show that FasL-induced pathway triggers expression of proinflammatory cytokines, including IL-1 beta, IL-6, MIP 2 and MCP-1, while GM-CSF-dependent pathway promotes functional maturation and activation of DCs. Taken together, the results indicate that dual gene-based delivery with FasL and GM-CSF may serve as a more effective tumor vaccine to suppress lung cancer cell growth in vivo. PMID- 18386792 TI - The ethics of advertising, billing, and finances in psychotherapy. AB - Psychotherapists must deal with practical business matters such as advertising, billing, collecting fees, and other practice management topics. We review the enforceable standards of the American Psychological Association's (2002) Ethics Code that deal with advertising, fees, billing, and related business matters in psychotherapy. Using a principle-based perspective, we link each of the standards to overarching ethical values and illustrate the concerns with case vignettes. We argue that understanding the moral foundations of ethical standards helps psychotherapists to implement with greater integrity the spirit and the letter of the standards with regard to advertising and business practices. PMID- 18386793 TI - Prehistoric trepanation in the Cuzco region of Peru: a view into an ancient Andean practice. AB - In this study, patterns of prehistoric trepanation in the southern highlands of Peru were examined through an analysis of 11 Cuzco-region burial sites. Trepanations were found in 66 individuals, with several individuals exhibiting more than one trepanation, for a total of 109 perforations observed. The predominant methods used were circular cutting and scraping-methods that proved highly successful with an overall 83% survival rate and little ensuing infection. Survival rates showed a significant increase over time, apparently reflecting improvements in trepanation technique through experimentation and practical experience. Practitioners avoided certain areas of the cranium and employed methods that reduced the likelihood of damage to the cerebral meninges and venous sinuses. In many cases, trepanation as a medical treatment appears to have been prompted by cranial trauma, a finding that corroborates other studies pointing to cranial trauma as a primary motivation for the surgical procedure. PMID- 18386794 TI - Using ancient mtDNA to reconstruct the population history of northeastern North America. AB - Mitochondrial DNA (mtDNA) was extracted and analyzed from the skeletal remains of 44 individuals, representing four prehistoric populations, and compared to that from two other prehistoric and several contemporary Native American populations to investigate biological relationships and demographic history in northeastern North America. The mtDNA haplogroup frequencies of ancient human remains from the Morse (Red Ocher tradition, 2,700 BP) and Orendorf (Mississippian tradition, 800 BP) sites from the Central Illinois River Valley, and the Great Western Park (Western Basin tradition, 800 BP) and Glacial Kame (2,900 BP) populations from southwestern Ontario, change over time while maintaining a regional continuity between localities. Haplotype patterns suggest that some ancestors of present day Native Americans in northeastern North America have been in that region for at least 3,000 years but have experienced extensive gene flow throughout time, resulting, at least in part, from a demic expansion of ancestors of modern Algonquian-speaking people. However, genetic drift has also been a significant force, and together with a major population crash after European contact, has altered haplogroup frequencies and caused the loss of many haplotypes. PMID- 18386795 TI - Early sexual maturity among Pume foragers of Venezuela: fitness implications of teen motherhood. AB - Because humans have slow life histories, discussions of the optimal age at first birth have stressed the benefits of delayed reproduction. However, given the diversity of ecological, fertility, and mortality environments in which humans live, reproductive maturity is expected to be highly variable. This article uses reproductive histories to examine a pattern of early menarche and first birth among the Pume, a group of South American foragers. Age at menarche and first birth are constructed using both retrospective and cross-sectional data for females over the age of 10 (n = 83). The objectives are first to define these patterns and then discuss their reproductive consequences. On average, Pume girls reach menarche at age 12.9, and give birth to their first child at age 15.3-15.5 (retrospective and cross-sectional data, respectively). This populational average falls several years prior to what often is considered the human norm. Two questions are then considered. What are the infant mortality costs across a mother's reproductive career? How does surviving fertility vary with age at first birth? Results indicate that the youngest of first-time mothers (<14) are four times more likely to loose their firstborns than older first-time mothers (> or =17). Given parity-specific mortality rates, the optimal strategy to minimize infant mortality and maximize reproductive span is to initiate childbearing in the midteens. Women gain no additional advantage in surviving fertility by delaying childbearing until their late teens. PMID- 18386796 TI - Mobility and subsistence economy: a diachronic comparison between two groups settled in the same geographical area (Liguria, Italy). AB - The purpose of this article is to investigate temporal shifts in skeletal robusticity to infer behavioral changes in two populations (Neolithic, NEOL and Medieval, MED) settled in the same geographic area but involved in different subsistence economies (pastoralism and coastal resources exploitation). This comparison allows us to test the hypothesis that occupational stress and mobility in the same environment produce predictable changes in the robusticity of both upper and lower limbs. Results show a lower degree of humeral robusticity and a similar degree of humeral asymmetry in the two sexes in the MED population. These results are consistent with the relatively less stressful subsistence economy documented in the MED population relative to that of the NEOL. Lower limb results suggest that femoral robusticity does not correlate directly with the level of logistical mobility, but is instead due to the summation of several diverse factors that place biomechanical loads on the hindlimb, particularly unevenness of the terrain. However, female femoral gracility seems to indicate that below a certain "threshold" of mobility, i.e., movement over the natural terrain, terrain conformation is no longer the main contributing factor to femoral robusticity. The femoral shape index I(x)/I(y) declines through time, particularly in males. This agrees with the expected mobility of the samples based on archaeological and historical data, providing further evidence on the reliability of this index in inferring terrestrial mobility. PMID- 18386797 TI - Problems in determination of skeletal lead burden in archaeological samples: an example from the First African Baptist Church population. AB - Human bone lead content has been demonstrated to be related to socioeconomic status, occupation and other social and environmental correlates. Skeletal tissue samples from 135 individuals from an early nineteenth century Philadelphia cemetery (First African Baptist Church) were studied by electrothermal atomic absorption spectrometry and X-ray fluorescence for lead content. High bone lead levels led to investigation of possible diagenetic effects. These were investigated by several different approaches including distribution of lead within bone by X-ray fluorescence, histological preservation, soil lead concentration and acidity as well as location and depth of burial. Bone lead levels were very high in children, exceeding those of the adult population that were buried in the cemetery, and also those of present day adults. The antemortem age-related increase in bone lead, reported in other studies, was not evidenced in this population. Lead was evenly deposited in areas of taphonomic bone destruction. Synchrotron X-ray fluorescence studies revealed no consistent pattern of lead microdistribution within the bone. Our conclusions are that postmortem diagenesis of lead ion has penetrated these archaeological bones to a degree that makes their original bone lead content irretrievable by any known method. Increased bone porosity is most likely responsible for the very high levels of lead found in bones of newborns and children. PMID- 18386798 TI - Some historical aspects of diabetic foot disease. AB - During the 19th century and for much of the 20th century, disease of the lower limb in diabetic patients was conceptualized not, as it is now, as 'the diabetic foot' or as 'a diabetic foot ulcer' but as 'gangrene in the diabetic foot' or as 'diabetic gangrene'. The prognostically and therapeutically important distinction between gangrene due to vascular insufficiency and gangrene due to infection in a limb with a normal or near normal blood supply was not made until about 1893. The advent of aseptic surgery improved the survival of amputation flaps, but surgery remained a hazardous undertaking until the discovery of insulin. Although insulin therapy reduced the risk of surgical intervention, diabetic foot disease now replaced hyperglycaemic coma as the major cause of diabetic mortality. The increasing workload attributable to diabetic foot disease after the introduction of insulin is reflected in the publications on diabetes in the 1920s. In some hospitals in North America this led to initiatives in prophylactic care and patient education, the importance of which were only more widely appreciated some 60 years later. A continuing emphasis on ischemia and infection as the major causes of diabetic foot disease led to a neglect of the role of neuropathy. In consequence, the management of diabetic neuropathic ulceration entered a prolonged period of therapeutic stagnation at a time when significant advances were being made in the management of lepromatous neuropathic ulceration. Reasons for the revival of progress in the management of diabetic neuropathic ulceration in the 1980s will be discussed. PMID- 18386799 TI - The diagnosis of Costello syndrome: nomenclature in Ras/MAPK pathway disorders. PMID- 18386800 TI - Sotos syndrome: antenatal presentation. AB - There is little published information regarding the clinical presentation of Sotos syndrome in pregnancy. In this report, we describe the antenatal presentation of a child subsequently diagnosed with Sotos syndrome by molecular analysis. The pregnancy was complicated by a positive maternal serum screen and abnormal ultrasound findings including macrocephaly, polyhydramnios and decreased fetal movements. This is the first report of an elevated Down syndrome risk in a pregnancy with confirmed Sotos syndrome. Sotos syndrome should be included in the differential diagnosis of newborns with a normal karyotype where the pregnancy has demonstrated an increased risk for Down syndrome by maternal serum screening, especially in the presence of supportive ultrasound findings. PMID- 18386801 TI - Tibial developmental field defect is the most common lower limb malformation pattern in VACTERL association. AB - VACTERL association is one of the most common recognizable patterns of human malformation and has been recently defined as a multiple polytopic developmental field defect. Limb anomalies are a key component of this condition and characteristically reflect perturbation of radial ray development. However, the pattern of appendicular malformations in VACTERL association is wider and includes a broad spectrum of additional and apparently nonspecific anomalies. We report on the sporadic case of a 4-10/12-year-old boy presenting with multiple costovertebral defects, dextrocardia, bilateral radial ray hypo/aplasia, unilateral kidney agenesis and anal atresia. Homolaterally to the more severe radial ray defect and kidney aplasia, he also has a complex lower limb malformation, consisting of distal tibial aplasia, clubfoot, hallucal deficiency and preaxial polydactyly. Literature review identifies 24 additional patients with VACTERL manifestations and lower limb malformations (excluding cases with isolated secondary deformations). Tibial hypo/aplasia with or without additional tibial field defects, reported in about 2/3 (68%) of the patients, represents the most common finding, while involvement of the fibular ray is rare (20%) and very often accompanies tibial anomalies. The relatively high frequency of tibial ray anomalies in VACTERL patients could easily be explained by the principle of homology of the developmental field theory. Careful search of lower limb anomalies of the "tibial type" is, therefore, indicated in all patients with multiple polytopic developmental field defects. PMID- 18386802 TI - Transposition of the great vessels in a patient with a 2.9 Mb interstitial deletion of 9q31.1 encompassing the inversin gene: clinical report and review. PMID- 18386803 TI - Omphalocele and gastroschisis and associated malformations. AB - The etiology of gastroschisis and omphalocele is unclear and their pathogenesis is controversial. Because previous reports have inconsistently noted the type and frequency of malformations associated with omphalocele and gastroschisis, we assessed these associated malformations ascertained between 1979 and 2003 in 334,262 consecutive births. Of the 86 patients with omphalocele, 64 (74.4%) had associated malformations. These included patients with chromosomal abnormalities (25, 29.0%); non-chromosomal syndromes including Beckwith-Wiedemann syndrome, Goltz syndrome, Marshall-Smith syndrome, Meckel-Gruber syndrome, Oto-palato digital type II syndrome, CHARGE syndrome, and fetal valproate syndrome; malformation sequences, including ectopia cordis, body stalk anomaly, exstrophy of bladder, exstrophy of cloaca, and OEIS (Omphalocele, Exstrophy of bladder, Imperforate anus, Spinal defect); malformation complexes including Pentalogy of Cantrell, and non-syndromic multiple congenital anomalies (MCA) (26, 30.2%). Malformations of the musculoskeletal system (31, 23.5%), urogenital system (27, 20.4%), cardiovascular system (20, 15.1%), and central nervous system (12, 9.1%) were the most common other congenital malformations in patients with omphalocele and non-syndromic MCA. Of the 60 patients with gastroschisis, 10 (16.6%) had associated malformations. In contrast to omphalocele, gastroschisis was rarely associated with a complex pattern of malformation, that is, one each (1.7%) with a chromosomal abnormality (trisomy 21), sequence (amyoplasia congenita), unspecified dwarfism, and 7 (11.7%) with MCA. We observed a striking difference in the prevalence of total malformations (74.4% vs. 16.6%, P < 0.001) and specific patterns of malformations associated with omphalocele and gastroschisis which emphasizes the need to evaluate all patients with omphalocele and gastroschisis for possible associated malformations. Malformation surveillance programs should be aware that the malformations associated with omphalocele can be often classified into a recognizable malformation syndrome or pattern (44.2%). PMID- 18386804 TI - Spondylocarpotarsal synostosis: long-term follow-up of a case due to FLNB mutations. PMID- 18386805 TI - Intrachromosomal partial triplication of chromosome 13 secondary to a paternal duplication with mild phenotypic effect. AB - Intrachromosomal triplications are rare and can be mistaken for duplications. The majority of triplications reported are de novo, mostly involving chromosome 15q, and have a middle inverted repeat. We report on the clinical, cytogenetic, and molecular analyses of a patient with a novel triplication 13q21.1-q21.33 secondary to a familial duplication 13q21.1-q21.33 with mild phenotypic effect in three generations. The propositus was an 8-year-old boy referred because of language delay and mild mental retardation. His weight, height and OFC were above the 97th centile. He had delayed tooth eruption and subtle dysmorphic features. Chromosome analysis (550 band stage) showed extra material in 13q21. Family history was unremarkable except for adult-onset sensorineural hearing loss in the father and paternal grandfather. Their karyotypes and those of both brothers of the propositus also showed an abnormal chromosome 13 but with less extra genetic material. FISH analysis with several BAC clones showed a triplication in the propositus between 204N9 and 184B18 (which mapped to 13q21.1 and 13q21.33, respectively) and a direct duplication for the same fragment (around 12 Mb) in the rest of the family members with the abnormal chromosome 13. The FISH signals did not show a middle inverted repeat. We describe the first intrachromosomal triplication 13q21.1-q21.33 derived from a paternal duplication. Meiotic instability in the transmission of a duplication has not been previously observed. Phenotypic variability may be explained by chromosomal non-penetrance or dosage critical loci located in the triplicate/duplicate segment. PMID- 18386807 TI - Rare interstitial deletion 9q31.2 to q33.1 de novo: longitudinal study in a patient over a period of more than 20 years. AB - The female carrier of a de novo interstitial deletion 9q [karyotype 46,XX,del(9)(q31.2q33.1)] was followed up over a period of more than 20 years. She shows facial dysmorphisms and significant growth retardation. Motor abilities are restricted by muscular hypotonia and malposition of the feet. She has mental retardation. There was no speech development and phases of autism were reported. By analyses with FISH and short tandem repeat markers, the interstitial deletion was confirmed and characterized to span 9q31.2q33.1, comprising at least 7.07 Mb. The aberration is of paternal origin. PMID- 18386806 TI - Mitochondrial ND5 T12338C, tRNA(Cys) T5802C, and tRNA(Thr) G15927A variants may have a modifying role in the phenotypic manifestation of deafness-associated 12S rRNA A1555G mutation in three Han Chinese pedigrees. AB - We report here on the clinical, genetic, and molecular characterization of three Han Chinese pedigrees with aminoglycoside-induced and nonsyndromic hearing loss. Clinical evaluation revealed the variable phenotype of hearing impairment including severity, age-at-onset, audiometric configuration in these subjects. The penetrance of hearing loss in WZD8, WZD9, and WZD10 pedigrees were 46%, 46%, and 50%, respectively, when aminoglycoside-induced deafness was included. When the effect of aminoglycosides was excluded, the penetrance of hearing loss in these pedigrees were 23%, 31%, and 37.5%, respectively. Mutational analysis of the complete mitochondrial genomes showed the homoplasmic A1555G mutation and distinct sets of mitochondrial DNA variants belonging to haplogroups D4b2b, B5b1, and F2, respectively. Of these, the tRNA(Cys) T5802C, tRNA(Thr) A15924C, and ND5 T12338C variants are of special interest as these variants occur at positions which are highly evolutionarily conserved nucleotides of tRNAs or amino acid of polypeptide. These homoplasmic mtDNA variants were absent among 156 unrelated Chinese controls. The T5802C and G15927A variants disrupted a highly conserved A U or C-G base-pairing at the anticodon-stem of tRNA(Cys) or tRNA(Thr), while the ND5 T12338C mutation resulted in the replacement of the translation-initiating methionine with a threonine, and also located in two nucleotides adjacent to the 3' end of the tRNA(Leu(CUN)). Thus, mitochondrial dysfunctions, caused by the A1555G mutation, would be worsened by these mtDNA variants. Therefore, these mtDNA mutations may have a potential modifier role in increasing the penetrance and expressivity of the deafness-associated 12S rRNA A1555G mutation in those Chinese pedigrees. PMID- 18386808 TI - Specific ultrasonographic features of perinatal lethal hypophosphatasia. AB - Prenatal diagnosis of perinatal lethal hypophosphatasia (PL-HPH) by ultrasonography is difficult as PL-HPH must be differentiated from other skeletal dysplasias with short long bones and poor mineralization of the skeleton, such as osteogenesis imperfecta type II and achondrogenesis/hypochondrogenesis. Here we present a case of molecularly confirmed PL-HPH and illustrate specific ultrasonographic findings that help to distinguish PL-HPH from similar conditions. PMID- 18386809 TI - Disruption of chromodomain helicase DNA binding protein 2 (CHD2) causes scoliosis. AB - Herein we characterize an apparently balanced de novo translocation, t(X;15)(p22.2;q26.1)dn, in a female patient with scoliosis, hirsutism, learning problems, and developmental delay (DGAP025). Other clinical findings include a high-arched palate, 2-3 syndactyly of the toes, and mildly elevated serum testosterone. No known or predicted genes are disrupted by the Xp22.2 breakpoint. The 15q26.1 breakpoint disrupts chromodomain helicase DNA binding protein 2 (CHD2). Another member of the chromatin-remodeling gene family, CHD7, has been associated with a defined constellation of congenital anomalies known as coloboma, heart anomaly, choanal atresia, mental retardation, genital and ear anomalies syndrome (CHARGE) and idiopathic scoliosis. Monosomy of 15q26 also has been associated with a spectrum of congenital abnormalities and growth retardation that overlaps with those of DGAP025. To provide a biological correlate, we characterized a mutant mouse model with Chd2 disruption that is associated with embryonic and perinatal lethality. Expression analysis indicated that Chd2 is expressed in the heart, forebrain, extremities, facial and dorsal regions during specific times of embryonic development. Chd2(+/m) mice showed pronounced lordokyphosis, reduced body fat, postnatal runting, and growth retardation. These data suggest that haploinsufficiency for CHD2 could result in a complex of abnormal human phenotypes that includes scoliosis and possibly features similar to CHARGE syndrome. PMID- 18386810 TI - Nutritional and genetic determinants of vitamin B and homocysteine metabolisms in neural tube defects: a multicenter case-control study. AB - Neural tube defects (NTDs) are severe congenital malformations due to failure of neural tube formation in early pregnancy. The proof that folic acid prevents NTDs raises the question of whether other parts of homocysteine (Hcy) metabolism may affect rates of NTDs. This French case-control study covered: 77 women aged 17-42 years sampled prior to elective abortion for a severe NTDs (cases) and 61 women aged 20-43 years with a normal pregnancy. Plasma and erythrocyte folate, plasma B6, B12 and Hcy were tested as five polymorphisms MTHFR 677 C --> T, MTHFR 1298 A --> C, MTR 2756 A --> G, MTTR 66 A --> G and TCN2 776 C --> G. Cases had significantly lower erythrocyte folate, plasma folate, B12 and B6 concentrations than the controls, and higher Hcy concentration. The odds ratio was 2.15 (95% CI: 1.00-4.59) for women with the MTRR 66 A --> G allele and it was decreased for mothers carrying the MTHFR 1298 A --> C allele. In multivariate analysis, only the erythrocyte folate concentration (P = 0.005) and plasma B6 concentration (P = 0.020) were predictors. Red cell folate is the main determinant of NTDs in France. Folic acid supplement or flour fortification would prevent most cases. Increased consumption of vitamins B12 and B6 could contribute to the prevention of NTDs. Genetic polymorphisms played only a small role. Until folic acid fortification becomes mandatory, all women of reproductive age should consume folic acid in a multivitamin that also contains B12 and B6. PMID- 18386812 TI - Page kidney after renal allograft biopsy: sonographic findings. AB - The development of renal failure and hypertension due to extrinsic renal compression by hematoma in the subcapsular space is well-described and most commonly seen after blunt trauma. It may also occur as a complication of percutaneous renal biopsy, both in native kidneys and renal allografts, and is a rare cause of reversible allograft failure. We describe a case of Page kidney in renal transplantation after percutaneous biopsy causing allograft dysfunction. Early recognition with sonography is important if irreversible damage to the allograft is to be prevented. PMID- 18386813 TI - Antenatal 3-dimensional sonographic diagnosis of fetal oropharyngeal teratoma. AB - We report a case of a large fetal oropharyngeal teratoma in a 19-week fetus evaluated with 3-dimensional (3D) sonography. The 3D sonographic surface and maximum mode rendering of the tumor allowed detailed visualization of the mass lesion and thus enabled active patient participation in the management of the pregnancy. PMID- 18386814 TI - A new prostate cancer therapeutic approach: combination of androgen ablation with COX-2 inhibitor. AB - Prostate cancer is initially responsive to hormonal therapy, but cancers inevitably progress in an androgen-independent fashion with virtually all tumors evolving into more aggressive androgen refractory disease. Immunohistological comparisons of cyclooxygenase 2 (COX-2) expressions in 3 pairs of prostate cancer patients before and after the combined androgen blockade (CAB) therapy show elevated COX-2 expressions. This observation from clinical specimens is further supported by in vitro laboratory data using human prostate cancer cells in which the antiandrogen hydroxyflutamide (HF) induced COX-2 expression, and androgen suppressed COX-2 expression. By applying knockdown and overexpression strategies to modulate AR expression in prostate cancer cells, we confirmed that androgen/AR signal suppressed, and HF induced COX-2 expression at both protein and mRNA levels. COX-2 promoter reporter assay indicated that the suppression of COX-2 by androgen/AR is at the transcriptional level via modulation of NF-kappaB signals. Treatment of LNCaP and LAPC4 cells with 1 microM HF in the presence of 1 nM DHT, which mimics the CAB therapy condition, promotes cell growth, and this growth induction can be suppressed via adding the COX-2 specific inhibitor, NS398. This suggests that HF promoted prostate cancer cell growth is COX-2 dependent and this HF-COX-2 activation pathway can account for one reason of CAB therapy failure. Together, these findings provide a possible explanation how CAB with antiandrogen HF therapy might fail and provide a potential new therapeutic approach to battle prostate cancer via combination of CAB therapy with COX-2 inhibitor(s). PMID- 18386815 TI - Expression and splicing of the unfolded protein response gene XBP-1 are significantly associated with clinical outcome of endocrine-treated breast cancer. AB - X-box binding protein 1 (XBP-1) is stimulated by endoplasmic reticulum stress as part of the unfolded protein response (UPR), which can promote apoptosis or cell survival. Non-conventional splicing, stimulated during the UPR, converts mRNA for "unspliced" XBP-1U to "spliced" XBP-1S mRNA. XBP-1 mRNA is oestrogen-responsive, but XBP-1S confers oestrogen independence and anti-oestrogen resistance to breast cancer cell lines. We therefore evaluated XBP-1 mRNA splicing as a factor in response of breast cancer patients to endocrine treatment. XBP-1 isoforms were measured by quantitative RT-PCR in 100 primary breast cancer patients treated with adjuvant tamoxifen (including 30 ER alpha-negative cases). In ER alpha positive cases, levels of XBP-1U mRNA correlated with ER alpha mRNA levels and were lower in grade 3 tumors. Higher levels of XBP-1U mRNA were significantly associated with breast cancer survival (Log-rank p = 0.002; Cox hazard ratio (HR) 0.2, p = 0.005), independent of grade, size, nodal status and progesterone receptor status. However, in the full cohort, higher ratios of XBP-1S/XBP-1U mRNA (indicating enhanced splicing) were associated with poor survival (Log-rank p = 0.03; Cox HR 2.3, p = 0.03) and related factors: ER alpha-negative status, progesterone receptor negative status, grade 3 tumors and greater proliferation. Significant associations with poor outcome were also seen for XBP-1 splicing in ER alpha-positive cases. Our findings, that XBP-1 isoforms are differently associated with outcome of endocrine therapy for patients, can be explained by higher levels of dominant-negative XBP-1U favouring apoptosis of tumor cells and higher levels of XBP-1S increasing tumor survival. PMID- 18386816 TI - EGFR tyrosine kinase inhibition radiosensitizes and induces apoptosis in malignant glioma and childhood ependymoma xenografts. AB - Malignant gliomas and childhood ependymomas have a high rate of treatment failure. Epidermal growth factor receptor (EGFR) activation has been implicated in the tumorigenesis and radioresistance of many cancers, including brain tumors. Therefore, combining EGFR targeting with irradiation is a potentially attractive therapeutic option. We evaluated the tyrosine kinase inhibitor gefitinib for its antitumor activity and potential to radio-sensitize in vivo in two xenograft models: an EGFR amplified glioma and an EGFR expressing ependymoma, both derived from primary tumors. When administered at 100 mg/kg for 5 consecutive days, gefitinib-induced partial tumor regression in all treated EGFR amplified IGRG88 glioma xenografts. The addition of 1 Gy of irradiation prior to gefitinib administration resulted in 5 complete and 4 partial regressions for the 9 treated tumors as well as a significant tumor growth delay of 33 days for the combined treatment compared to 19 days for each therapy alone, suggesting additive antitumor activity. Tumor regression was associated with inhibition of AKT and MAPK pathways by gefitinib. In contrast, the ependymoma IGREP83 was sensitive to irradiation, but remained resistant to gefitinib. Combined treatment was associated with inhibition of radiation-induced MAPK phosphorylation and significant induction of apoptotic cell death though radiation-induced AKT phosphorylation was maintained. Depending on the scheduling of both therapies, a trend towards superior antitumor activity was observed with combined treatment. Thus, EGFR targeting through tyrosine kinase inhibition appears to be a promising new approach in the treatment of EGFR-driven glioma, particularly in combination with radiation therapy. PMID- 18386817 TI - DNA repair deficiency leads to susceptibility to develop arsenic-induced premalignant skin lesions. AB - In West Bengal, India, although more than 6 million people are exposed to arsenic through drinking water, only 15-20% showed arsenic-induced skin lesions, including premalignant hyperkeratosis. This indicates toward some factors that confer susceptibility to arsenic-induced carcinogenicity. In this work, we wanted to explore whether differences in DNA repair capacity could impart arsenic induced carcinogenicity, through Comet assay, chromosomal aberration (CA) assay and challenge assay. Sixty arsenic exposed (30 individuals with arsenic-induced premalignant hyperkeratosis and 30 without skin lesion, but drinking similar arsenic contaminated water) and 30 arsenic unexposed individuals were recruited as study participants. Alkaline comet assay, and challenge assay were carried out in whole blood and CA study in lymphocytes to find out the DNA damage and DNA repair capacity in both hyperkeratotic and without skin lesion individuals. DNA damage as well as CA were found to be significantly higher in the arsenic-exposed individuals compared to unexposed individuals (p < 0.001). Within the exposed group, there was no significant difference as far as the level of DNA damage is concerned (p > 0.05), but CA was significantly higher in exposed individuals with hyperkeratosis than exposed individuals without hyperkeratosis (p < 0.01). Challenge assay showed that upon induction of DNA damage, the repair capacity in the exposed individuals with premalignant hyperkeratosis is significantly less (p < 0.001) than that of individuals without skin lesion, although the basal level of DNA damage was similar in both. Thus, the deficiency in DNA repair capacities in the hyperkeratotic individuals emerges as a prime contender for arsenic carcinogenicity. PMID- 18386818 TI - Reduced expression of IL-18 is a marker of ultraviolet radiation-induced melanomas. AB - We previously showed that mice carrying an activated Cdk4 mutation together with melanocyte-specific mutant Hras (Cdk4(R24C/R24C)/TPras) develop melanoma spontaneously, but penetrance is increased and age of onset reduced after neonatal ultraviolet radiation (UVR) exposure. UVR-treated mice were more likely to develop multiple primary lesions, and these melanomas more often expressed Trp53, and less often expressed c-Myc, than melanomas from nonirradiated mice (Hacker et al., Cancer Res 2006;66:2946-52). These data suggest differences in mechanisms of tumorigenesis between melanomas developing spontaneously, or as a result of UVR exposure. To further delineate these differences, we compared global gene expression between spontaneous and UVR-induced melanomas from these mice using microarrays. We found 264 genes differentially expressed between these groups (ANOVA, p < 0.05). Selected candidate genes were validated using qRT-PCR, which confirmed upregulation of Gpr155 and Bmp7, and downregulation of Plagl1, Akap12 and Il18 in UVR-induced mouse melanomas. In humans, epidemiological studies suggest that there may be 2 predominant pathways to melanoma development. One characterized by chronic UVR exposure and which leads mainly to melanomas on sun-exposed sites; the other associated with low UVR exposure and leading predominantly to melanomas on less-exposed body sites. We found by immunohistochemical analysis that, comparing a series of human melanomas from the head (a chronically sun-exposed site; N = 82) with a set from the trunk (an intermittently exposed site; N = 65), the prevalence of IL-18 expression was significantly lower in melanomas on the head (16%) than on truncal melanomas (34%, p = 0.011). We conclude that loss of IL-18 is a marker of UVR-induced melanoma, both in animal models and humans. PMID- 18386819 TI - Evolution of the thyroid: anterior-posterior regionalization of the Oikopleura endostyle revealed by Otx, Pax2/5/8, and Hox1 expression. AB - The thyroid in vertebrates and its homolog, the endostyle in nonvertebrate chordates, share a molecular code for dorsoventral patterning. Little is yet known, however, about mechanisms that pattern the endostyle's anterior-posterior (AP) axis. To extend our understanding of thyroid development and evolution, we studied Oikopleura dioica, a larvacean urochordate that retains a chordate body plan as adults. Transcription factor expression domains revealed AP regionalization of the endostyle, with expression of Otx rostrally, Hox1 caudally, and two Pax2/5/8 paralogs centrally. Comparative analysis suggested that the endostyle of stem chordates expressed orthologs of these genes and that ancestral subfunctions partitioned differentially among lineages. Because the ordered expression of Otx, Pax2/5/8, and Hox1 displays patterning in both the endodermally derived endostyle and the ectodermally derived central nervous system, we propose that this gene set belonged to the developmental genetic toolkit of stem bilaterians and repeatedly provided AP positional information in various developmental situations. PMID- 18386820 TI - Hsp60C is required in follicle as well as germline cells during oogenesis in Drosophila melanogaster. AB - Hsp60C gene of Drosophila melanogaster shows a dynamic spatiotemporal expression during oogenesis and seems to contribute bulk of the Hsp60 family proteins in ovarioles. Hsp60 distribution overlaps with that of F-actin-rich membranes/structures in follicle, nurse, and egg cells throughout oogenesis. Skeletal muscle fibers associated with ovarioles and in other parts of the body show patterned location of Hsp60 in A-bands. During stages 11-12, Hsp60 accumulates at junctions of nurse cells and oocyte, where a new microtubule organizing center is known to develop. A recessive hypomorph allele, Hsp60C1 causes complete sterility of the rare surviving homozygous adults. Their egg chambers show very little Hsp60C transcripts or Hsp60 protein. Beginning at stages 6-7, Hsp60C1 chambers show a disorganized follicle cell layer with poor cell adhesion in addition to abnormal organization of F-actin and other cytoskeletal structures in follicle, nurse, and egg cells. Additionally, expression and localizations of Hrb98DE, Squid, and Gurken proteins in nurse cells and oocyte are also severely affected. Hsp60C1 homozygous follicle cell clones in Hsp60C1/+ ovarioles show disruptions in follicle epithelial and cytoskeleton arrangements. Likewise, egg chambers with Hsp60C1 homozygous germline clones in Hsp60C1/+ flies show abnormal oogenesis. Our results provide the first evidence for an essential role of Hsp60C in Drosophila oogenesis, especially in organization and maintenance of cytoskeletal and cell adhesion components. PMID- 18386821 TI - Expression and regulation of the decoy bone morphogenetic protein receptor BAMBI in the developing avian face. AB - Here, we examine the expression and regulation of the gene BAMBI, a kinase deficient decoy receptor capable of interacting with type I bone morphogenetic protein (BMP) receptors in avian embryos. Initially, expression was limited to the endoderm during neurula and pharyngula stages. From embryonic day 3.5 (stage 20) and onward, BAMBI expression almost perfectly overlapped with known expression patterns for BMP4, particularly in the face and limbs. We performed bead implant experiments in the face to see which signals could be repressing or promoting expression of BAMBI. Our data point to retinoids and BMPs as being major positive regulators of BAMBI expression; however, fibroblast growth factor 2 acts to repress BAMBI. Furthermore, retinoic acid is likely to act directly on BAMBI as induction occurs in the presence of cycloheximide. The data suggested that BAMBI could be used to regulate Bmp signaling during tissue interactions that are an integral part of facial morphogenesis. PMID- 18386823 TI - Sonographic diagnosis of a cervical vagal schwannoma. AB - This case report illustrates the role of high-resolution sonography in the preoperative assessment of a schwannoma of the vagus nerve in the neck. Sonography identified the tumor in the right carotid space and determined its origin from the right vagus nerve, facilitating the surgeon's approach to preserve nerve function. PMID- 18386822 TI - PAX6 dosage effects on corneal development, growth, and wound healing. AB - The requirement for correct dosage of the transcription factor Pax6 during corneal growth and development was investigated using the Pax6-overexpressing (PAX77) transgenic mouse. Transgenics had a microcornea phenotype due to failure of postnatal growth, associated with reduction in the number of cells layers in the corneal epithelium. Cell cycle progression was monitored using bromodeoxyuridine, p63, cyclin E, and phosphohistone-3 labeling: proliferation rates were higher in PAX77+ than wild-type, without a concomitant increase in apoptosis. Hence, failure of proliferation did not underlie microcornea. PAX77+ corneal epithelia had reduced levels of cytokeratin-12, and exhibited severe wound healing delay that, in contrast to Pax6+/- mice, could not be modulated by exogenous growth factors. PAX77+ lenses showed partial failure of lens fiber differentiation. The data demonstrate that anterior eye development is very sensitive to Pax6 dosage. Although there are similarities between the eye phenotype of Pax6 heterozygotes and overexpressing mice, there are also striking differences. Developmental PMID- 18386824 TI - Fetal meconium peritonitis complicated with bacterial infection. AB - Fetal meconium peritonitis complicated by bacterial infection is extremely rare. We report a case of fetal ascites at 21 weeks of gestation with subsequent development of loculation, encapsulation, and calcification at 25 weeks. Paracentesis of loculated ascitic fluid at 28 weeks of gestation showed a purulent appearance with the presence of cocci bacteria, increase in white cell count, and a low glucose level, which were suggestive of bacterial infection. However, no sources of maternal infection could be identified. The total bilirubin level of the ascitic fluid was normal (21 micromol/L). A healthy baby was delivered at 37 weeks. CT scan revealed normal bowel without any sign of perforation. We postulate that when ascitic fluid becomes loculated, a normal bilirubin level on paracentesis indicates spontaneous closure of a previous bowel perforation. PMID- 18386825 TI - Pushing the envelope: developmental regulation by the nuclear lamina. AB - A surprising realization in recent years was that proteins of the nuclear lamina directly regulate gene expression and cell differentiation. Presented here are examples that highlight the diverse roles these proteins can inhabit. Accompanying the primer is a discussion of current topics in the field with nuclear lamina experts Colin Stewart, Ph.D., and Yosef Gruenbaum, Ph.D. PMID- 18386826 TI - The role of FGF signaling in the establishment and maintenance of mesodermal gene expression in Xenopus. AB - FGF signaling is important for the formation of mesoderm in vertebrates, and when it is perturbed in Xenopus, most trunk and tail mesoderm fails to form. Here we have further dissected the activities of FGF in patterning the embryo by addressing its inductive and maintenance roles. We show that FGF signaling is necessary for the establishment of xbra expression in addition to its well characterized role in maintaining xbra expression. The role of FGF signaling in organizer formation is not clear in Xenopus. We find that FGF signaling is essential for the initial specification of paraxial mesoderm but not for activation of several pan-mesodermal and most organizer genes; however, early FGF signaling is necessary for the maintenance of organizer gene expression into the neurula stage. Inhibition of FGF signaling prevents VegT activation of specific mesodermal transcripts. These findings illuminate how FGF signaling contributes to the establishment of distinct types of mesoderm. PMID- 18386827 TI - Erasure of the paternal transcription program during spermiogenesis: the first step in the reprogramming of sperm chromatin for zygotic development. AB - Male germ cells possess a unique epigenetic program and express a male-specific transcription profile. However, when its chromatin is passed onto the zygote, it expresses an transcription/epigenetic program characteristic of the zygote. The mechanism underlying this reprogramming process is not understood at present. In this study, we show that an extensive range of chromatin factors (CFs), including essential transcription factors and regulators, remodeling factors, histone deacetylases, heterochromatin-binding proteins, and topoisomerases, were removed from chromatin during spermiogenesis. This process will erase the paternal epigenetic program to generate a relatively naive chromatin, which is likely to be essential for installation of the zygotic developmental program after fertilization. We have also showed that transcription termination in male germ cells was temporally correlated with CF dissociation. A genome-wide CF dissociation will inevitably disassemble the transcription apparatus and regulatory mechanism and lead to transcription silence. Based on data presented in this and previous studies (Sun et al., Cell Research [2007] 17:117-134), we propose that paternal-zygotic transcription reprogramming begins with a genome wide CF dissociation to erase the existing transcription program in later stages of spermatogenesis. This will be followed by assembling of the zygotic equivalent after fertilization. The transcription/epigenetic program of the male germ cell is transformed into a zygotic one using an erase-and-rebuild strategy similar to that used in the maternal-zygotic transition. It is also noted that transcription is terminated long after meiosis is completed and before chromatin becomes highly condensed during spermatogenesis. The temporal order of these events suggests that transcription silence does not have to be coupled to meiosis or chromatin condensation. PMID- 18386828 TI - Is the increase in orchiectomy for prostate cancer patients appropriate? PMID- 18386829 TI - Heart failure associated with sunitinib malate: a multitargeted receptor tyrosine kinase inhibitor. AB - BACKGROUND: Sunitinib malate is a novel multitargeted receptor tyrosine kinase inhibitor with established efficacy in the treatment of metastatic renal cell carcinoma and imatinib-resistant gastrointestinal stromal tumor. This report describes the development of heart failure in cancer patients who received this novel agent. METHODS: A retrospective study was conducted at M. D. Anderson Cancer Center during a 1-year period on patients who received sunitinib and developed heart failure. RESULTS: During 2006, 6 of 224 (2.7%) patients who received sunitinib developed heart failure (HF) that resulted in substantial morbidity and, in some cases, mortality. Symptomatic heart failure occurred soon after initiation of sunitinib (mean onset 22 days after initiation), was associated with decline in cardiac function and elevations in blood pressure, and was not completely reversible in most patients, even after termination of sunitinib therapy. CONCLUSIONS: These observations suggested that sunitinib associated heart failure may represent a potentially serious toxicity and underscore the need for careful monitoring of cardiac function and aggressive control of hypertension in these patients. Studies to elucidate potential mechanisms of heart failure and left ventricular dysfunction resulting from treatment with sunitinib are necessary to develop strategies for prevention and treatment of this complication. PMID- 18386831 TI - Development of a bioactive glass fiber reinforced starch-polycaprolactone composite. AB - For bone regeneration and repair, combinations of different materials are often needed. Biodegradable polymers are often combined with osteoconductive materials, such as bioactive glass (BaG), which can also improve the mechanical properties of the composite. The aim of this work was to develop and characterize BaG fiber reinforced starch-poly-epsilon-caprolactone (SPCL) composite. Sheets of SPCL (30/70 wt %) were produced using single-screw extrusion. They were then cut and compression-molded in layers with BaG fibers to form composite structures with different combinations. Mechanical and degradation properties of the composites were studied. The actual amount of BaG in the composites was determined using combustion tests. Initial mechanical properties of the reinforced composites were at least 50% better than the properties of the nonreinforced specimens. However, the mechanical properties of the composites after 2 weeks of hydrolysis were comparable to those of the nonreinforced samples. During the 6 weeks hydrolysis the mass of the composites had decreased only by about 5%. The amount of glass in the composites remained as initial for the 6-week period of hydrolysis. In conclusion, it is possible to enhance initial mechanical properties of SPCL by reinforcing it with BaG fibers. However, mechanical properties of the composites are typical for bone fillers and strength properties need to be further improved for allowing more demanding bone applications. PMID- 18386830 TI - Comparison of Akt/mTOR signaling in primary breast tumors and matched distant metastases. AB - BACKGROUND: The Akt/mammalian target of the rapamycin (mTOR) signaling pathway represents a promising target for cancer therapy. The phosphorylation status of Akt and of mTOR's phosphorylation target eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) is often used to assess the activity of Akt and mTOR signaling. The purpose of this study was to determine whether primary tumors differ from their metastasis in their expression of pAkt and p4E-BP1. METHODS: Primary breast tumors and their distant metastases surgically resected from the same patients were evaluated with immunohistochemical analysis (IHC) for pAkt (Ser473) and p4E-BP1 (Ser65). The agreement between the IHC results for the primary tumor and metastases was evaluated with Cohen kappa (kappa). RESULTS: Most primary breast tumors and metastatic tumors expressed pAkt (76% of each). Of the 23 matched evaluable pairs, however, 11 (47.8%) had discordant IHC results (kappa -0.31; 95% confidence interval [CI], -0.49 to -0.13). Similarly, although most of the primary and metastatic tumors were positive for p4E-BP1 (75% and 74%), of the 23 matched evaluable pairs, 8 (47.8%) were discordant (kappa 0.10; 95% CI, -0.33-0.52). CONCLUSIONS: In this series, most primary breast tumors and metastases expressed pAkt and p4E-BP1 by IHC. Concordance between IHC findings in primary tumors and metastases was poor, however. Further work is needed to determine whether this reflects true biological heterogeneity or poor reproducibility of IHC with phosphospecific antibodies, and to identify which biomarkers can be assessed most reproducibly in primary tumors to predict activity of Akt/mTOR signaling and sensitivity to pathway inhibitors. PMID- 18386832 TI - The effect of temperature and initial pH on biomimetic apatite coating. AB - Bone-like apatite coatings were prepared using a biomimetic method in a simulated body fluid (SBF). The effect of initial pH values and immersing temperatures on biomimetic apatite coating formation was studied. Three different temperatures were used in this study: 24 (room temperature), 40, and 60 degrees C. At each temperature, SBF solutions with three different initial pHs were chosen: low, medium, and high. The total inorganic carbon (TIC) content and pH-time profile of each coating system were recorded during the coating formation. The apatite coatings were characterized using X-ray diffraction (XRD), field emission scanning electron microscope (FESEM), and Fourier transform infra-red (FTIR). It has been found that SBF temperature has a great effect on the bicarbonate decomposition rate. The bicarbonate ions tend to decompose faster as the temperature increases. The decomposition of bicarbonate ions results in a pH increase in the SBF. With different initial SBF pHs, the decomposition of different amounts of bicarbonate ions is required to reach the critical pH range of apatite formation. With different amounts of bicarbonate ions in the SBF, the surface morphology of the biomimetic apatite coating formed is different. Therefore, the initial pH of the SBF solution plays a vital role in controlling the surface morphology of the biomimetic apatite coating. Also, it was found that as the SBF temperature increased, the critical pH range at which biomimetic apatite coating forms decreased. The critical pH range for the SBF prepared at 24, 40, and 60 degrees C was 6.65-6.71, 6.55-6.65, and 6.24-6.42, respectively. PMID- 18386833 TI - Self-assembling peptide nanofibers and skeletal myoblast transplantation in infarcted myocardium. AB - Cell transplantation is currently limited by poor graft retention and survival in the postinfarction scar. Because this issue could potentially be addressed by embedding cells in bioinjectable scaffolds and boosting cell survival pathways, we induced a myocardial infarction in 72 rats to assess the effects of different self-assembling peptides with or without platelet-derived growth factor (PDGF-BB) on survival of transplanted skeletal myoblasts. Two weeks after coronary artery ligation, rats were randomized to receive in-scar injections of culture medium (controls, n = 11), self-assembling peptide (RAD16-I) nanofibers (NF, n = 9), skeletal myoblasts (n = 12), or skeletal myoblasts in combination with NF (n = 8). In separate experiments with different self-assembling peptides (RAD16-II), rats received in-scar injections of culture medium (controls, n = 6), skeletal myoblasts (n = 10), PDGF-loaded peptides (n = 7), or skeletal myoblasts (5 x 10(6)) in combination with PDGF-loaded peptides (n = 9). After 1 month, left ventricular function, as assessed by echocardiography, was not improved in either of the experimental groups compared with controls. This correlated with the failure of RAD16-I peptides or PDGF-loaded RAD16-II peptides to improve myoblast survival despite a greater angiogenesis. In vitro experiments confirmed that the number of myoblasts decreased over time when seeded on nanofiber gels. These data suggest that the optimal use of biomaterial scaffolds for survival of transplanted cells will require specific tailoring of the biomaterial to the cell type. PMID- 18386834 TI - Surface modification of polycaprolactone membrane via layer-by-layer deposition for promoting blood compatibility. AB - The polycaprolactone (PCL) membranes were successfully modified by deposition of chitosan/heparin multilayer via a simple electrostatic self-assembly method. To immobilize chitosan, a novel ternary polysaccharide derivate, chitosan-g-PCL-b poly-(ethylene glycol) (PEG) was used coating on PCL film first, which resulted in the presence of positive charges onto PCL surface as the basis for following electrostatic self-assembly. The process of modification was monitored by X-ray photoelectron spectroscopy, static contact angle measurement, and atomic force microscopy. The chitosan-g-PCL-b-PEG/heparin complex immobilized on PCL surface presented it with increasing hydrophilicity and microphase separation structure. Then in vitro hemocompability experiments indicated that this multilayer deposition on PCL resisted the platelets adhesion and prolonged the plasma recalcification time effectively, relative to the untreated PCL. Such chitosan-g PCL-b-PEG/heparin-modified PCL may have good potential for use in vascular tissue engineering. PMID- 18386835 TI - A practical approach to boundaries in psychotherapy: making decisions, bypassing blunders, and mending fences. AB - Nonsexual boundary crossings can enrich psychotherapy, serve the treatment plan, and strengthen the therapist-client working relationship. They also can undermine the therapy, disrupt the therapist-patient alliance, and cause harm to clients. Building on T. G. Gutheil and G. O. Gabbard's (1993) conceptualization of boundary crossings and boundary violations, this article discusses and illustrates grounding boundary decisions in a sound approach to ethics. We provides nine useful steps in deciding whether to cross a boundary, describe common cognitive errors in boundary decision making, and offer nine helpful steps to take when a boundary crossing has negative effects. PMID- 18386836 TI - Mechanistic study of electroosmotic transport across hydrated nail plates: effects of pH and ionic strength. AB - The objective of this study was to investigate the effects of pH and ionic strength on electroosmotic transport in transungual iontophoresis. Transungual iontophoretic transport of model neutral permeants mannitol (MA) and urea (UR) across fully hydrated human nail plates in phosphate-buffered saline of different pH and ionic strengths were investigated in vitro. Two protocols were involved in the transport experiments with each protocol divided into stages including passive and iontophoresis transport at 0.1 and/or 0.3 mA. Nail plate electrical resistance and water uptake of nail clippings were measured at various pH and ionic strengths. In the pH study, electroosmosis enhanced the anodal transport of MA at pH 9 and cathodal transport at pH 3. The Peclet numbers of MA were more than two times higher than those of UR under these conditions. No significant electroosmosis enhancement was observed for MA and UR at pH 5. In the ionic strength study, a decrease in solution ionic strength from 0.7 to 0.04 M enhanced electroosmotic transport. Nail electrical resistance increased with decreasing the ionic strength of the equilibrating solution, but reached a plateau when the ionic strength was less than approximately 0.07 M. Solution pH and ionic strength had no significant effect on nail hydration. Under the studied pH and ionic strength conditions, the effects of electroosmosis were small compared to the direct-field effects in transungual iontophoretic transport of small to moderate size permeants. PMID- 18386837 TI - Investigating improving powder deagglomeration via dry powder inhalers at a low inspiratory flow rate by employing add-on spacers. AB - The aim of this study was to investigate whether small add-on spacers alone or equipped with a passive deagglomerating component would improve aerosol performances of passive low airflow resistance dry powder inhalers (DPIs) at a low inhalation flow rate. Depositions of beclometasone dipropionate (BDP) and salbutamol sulphate (SS) via the Asmabec Clickhaler and Asmasal Clickhaler at 30 L/min airflow rate in an oropharyngeal model and attached filter were determined. Three add-on spacers, one with 5.0 cm length, and the other with the same features but incorporating a fine mesh, and the third one with the length of 8.5 cm (long add-on spacer) were used. Incorporating mesh did not improve the filter dose for SS, and significantly reduced this dose for BDP. The long add-on spacer was the most efficient spacer as it had minimal effects on the filter doses, also significantly reduced drug depositions in the model. In conclusion, an optimum length of an add-on spacer is required to minimise oropharyngeal drug deposition via a low airflow resistance DPI at a low inspiratory flow rate without considerable reduction of the respirable dose. Incorporating sieves within add-on spacers may diminish aerosol performances of the DPIs at low airflow rates. PMID- 18386838 TI - Site-specific drug delivery to the middle-to-lower region of the small intestine reduces food-drug interactions that are responsible for low drug absorption in the fed state. AB - Food-drug interactions may reduce the bioavailability of drugs taken after meals (negative food effects). We designed enteric-coated tablets that start to disintegrate when they reach the middle-to-lower region of the small intestine, and examined whether they could reduce negative food effects in dogs. Tablets containing trientine as a model drug were coated with hypromellose acetate succinate (HPMCAS) with various values of succinoyl group content. The time lag of drug dissolution from these enteric-coated tablets in simulated intestinal fluid of pH 6.8 increased as the succinoyl group content was decreased. The AUC of trientine after oral administration of its aqueous solution to fed dogs was one-eighth of that in fasted dogs. The low drug absorption in fed dogs was improved when trientine was administered as enteric-coated tablets. The average ratio of AUC in the fed state to that in the fasted state increased with decreasing succinoyl group content of HPMCAS. Negative food effects completely disappeared after oral administration of tablets coated with HPMCAS having a succinoyl group content of 6.2% or less, which probably disintegrated in the middle-to-lower small intestine. Our results indicated that food-drug interactions were avoided by separating the main absorption site of drugs from that of food components. PMID- 18386839 TI - Efficacy of titanium dioxide photocatalyst for inhibition of bacterial colonization on percutaneous implants. AB - The purpose of this study was to evaluate the efficacy of titanium dioxide photocatalyst in inhibition of bacterial colonization on percutaneous implants. Titanium dioxide photocatalyst was prepared by direct oxidization of pure titanium substrate, and a comparative study with pure titanium was performed. The bactericidal ability of the photocatalyst was examined using methicillin resistant Staphylococcus aureus (MRSA) suspensions in a colony-forming assay according to the Japanese Industrial Standards committee standard. After exposing the MRSA suspension on sample plates to ultraviolet A (UVA) light, the number of surviving bacteria was estimated. Next, an animal model for inhibition of colonization was examined in vivo. Pins were inserted into the femurs of rabbits, were infected with 10(8) colony-forming units of MRSA suspension, and were illuminated with UVA light for 60 min daily; the number of colonizing bacteria was estimated after 7 days. The bactericidal ability of the photocatalyst was apparent after 60 min, when the bacteria had almost disappeared. The number of colonizing bacteria on photocatalytic pins was decreased significantly in vivo. The photocatalyst was effective even against resistant bacterial colonization. Clinically, the incidence of percutaneous implant infection such as pin tract infection in external fixation could be reduced using the titanium photocatalyst. PMID- 18386840 TI - In vivo preclinical efficacy of a PDLLA/PGA porous copolymer for dental application. AB - This study aimed to analyze surface morphology and physical-chemical properties of a copolymer of polylactic/polyglycolic acid (Fisiograft, Ghimas SpA, Casalecchio di Reno, Italy) by scanning electron microscopy (SEM), porosimetry, and rheological analysis. Then the material was implanted in vivo to test its efficacy at promoting bone healing and new bone formation in postextraction sockets. Under general anaesthesia, sockets were created in 12 minipigs and then randomly filled with the porous copolymer in SPONGE or GEL form and compared with commercial BioOss (Geistlich Biomaterials) and Biocoral (Inoteb, France). At 15, 30, and 60 days from surgery, the newly formed trabecular bone quality was evaluated by means of histology and histomorphometry. The SEM and rheological analyses performed on GEL showed a surface microporosity and a rheological shear thinning behavior, whereas the SPONGE porosimetric measurements revealed larger pores. At 15 days, the new bone regrowth was observed in all treated sockets but appeared immature, as the trabeculae were very dense and thin. At 30 days, GEL and SPONGE were degraded, and the sockets were filled with bone that, in terms of bone volume fraction, trabecular number, and separation, was not statistically different from normal bone. PMID- 18386841 TI - Integrity of the stem-cement interface in THA: effects of stem surface finish and cement porosity. AB - No consensus exists for the optimal surface finish on cemented total hip prosthesis stems. The purpose of this study was to determine the effects of stem finish and interfacial cement porosity on the integrity of the stem-cement interface. Simulated stems made of Co-Cr, having polished or matte surfaces, at room temperature or heated to 37 degrees C, were cemented into Sawbones simulated femurs. Push out testing of the stem-cement interface was performed immediately after cement polymerization and after two aging periods in 37 degrees C saline or 37 degrees C air, and the extent of interfacial porosity at the stem-cement interface was determined. Polished stems exhibited an average 60% greater interfacial strength than that of matte stems initially and up to 240% after aging treatments. Cement porosity at the stem-cement interface and incomplete cement interdigitation into surface asperities on matte stems likely allowed saline penetration into the stem-cement interface during wet aging, resulting in a rapid decrease of shear strength. Stem preheating to 37 degrees C virtually eliminated interfacial pores and resulted in greater shear strengths regardless of surface finish. Polished stem surfaces with stem preheating provided the best interfacial shear strength and sealing ability against saline penetration into the stem-cement interface and could result in increased longevity of stem fixation. PMID- 18386842 TI - Evaluation of substrates for 90 degrees peel adhesion--a collaborative study. I. Medical tapes. AB - As part of a method development for peel testing, an interlaboratory comparison among Food and Drug Administration-Center for Drug Evaluation and Research, Food and Drug Administration-Center for Devices and Radiological Health and Southwest Research Institute was conducted using medical tapes. The aim was to determine which readily available substrate [stainless steel (SS), high density polyethylene (HDPE) or Vitro-Skin(R)] would best distinguish among various medical tapes. Five medical tapes (3M 1523, 3M 1525L, 3M 1776, Mepiform(R) and Mediderm(R) 3505) were evaluated on four different substrates (SS, HDPE, Vitro Skin, and human cadaver skin) using the following peel parameters: approximately 3 min dwell time, 90 degrees peel angle, and 300 mm/min peel rate. No substrate mimics cadaver skin for all five tapes. SS had the best ability to distinguish among the medical tapes. Overall, for quality control purposes (yielding good discrimination and precision), SS would be the optimal substrate. PMID- 18386843 TI - Guide to collagen characterization for biomaterial studies. AB - The structure and remodeling of collagen in vivo is critical to the pathology and healing of many human diseases, as well as to normal tissue development and regeneration. In addition, collagen matrices in the form of fibers, coatings, and films are used extensively in biomaterial and biomedical applications. The specific properties of these matrices, both in terms of physical and chemical characteristics, have a direct impact on cellular adhesion, spreading, and proliferation rates, and ultimately on the rate and extent of new extracellular matrix formation in vitro or in vivo. In recent studies, it has also been shown that collagen matrix structure has a major impact on cell and tissue outcomes related to cellular aging and differentiation potential. Collagen structure is complex because of both diversity of source materials, chemistry, and structural hierarchy. With such significant impact of collagen features on biological outcomes, it becomes essential to consider an appropriate set of analytical tools, or guide, so that collagens attained from commercial vendors are characterized in a comparative manner as an integral part of studies focused on biological parameters. The analysis should include as a starting point: (a) structural detail-mainly focused on molecular mass, purity, helical content, and bulk thermal properties, (b) chemical features-mainly focused on surface elemental analysis and hydrophobicity, and (c) morphological features at different length scales. The application of these analytical techniques to the characterization of collagen biomaterial matrices is critical in order to appropriately correlate biological responses from different studies with experimental outcomes in vitro or in vivo. As a case study, the analytical tools employed for collagen biomaterial studies are reviewed in the context of collagen remodeling by fibroblasts. The goal is to highlight the necessity of understanding collagen biophysical and chemical features as a prerequisite to (a) studies with cells and tissue formation, and (b) suggest modes to establish comparative outcomes for studies conducted in different laboratories. PMID- 18386844 TI - Prestresses in bilayered all-ceramic restorations. AB - INTRODUCTION: A general trend in all ceramic systems is to use veneering ceramics of slightly lower thermal expansion coefficients compared with that of the framework resulting in a positive mismatch in thermal expansion coefficient (+DeltaTEC). The concept behind this TEC mismatch is to generate compressive stresses in the weaker veneering ceramic and thus enhance the overall strength of the restoration. This technique had excellent results with porcelain fused to metal restorations (PFM). However, there are concerns to apply this concept to all-ceramic restorations. The aim of this research was to determine the stresses in bilayered all-ceramic restorations due to the mismatch in TEC. MATERIALS AND METHODS: Two commercial veneering ceramics with a TEC lower than that of zirconia (+DeltaTEC); NobelRondo zirconiatrade mark and Lava Ceramtrade mark, plus one experimental veneering ceramic with an identical TEC that matches that of zirconia (DeltaTEC = 0) were used to veneer zirconia discs. The specimens were loaded in biaxial flexure test setup with the veneer ceramic in tension. The stresses due to load application and TEC mismatch were calculated using fractography, engineering mathematics, and finite element analysis (FEA). RESULTS: In this study, the highest load at failure (64 N) was obtained with the experimental veneer where the thermal mismatch between zirconia and veneering ceramic was minimal. For the two commercial veneer ceramics the magnitude of the thermal mismatch localized at the zirconia veneer interface (42 MPa) exceeded the bond strength between the two materials and resulted in delamination failure during testing (ca. 50 MPa). SIGNIFICANCE: For all-ceramic zirconia veneered restorations it is recommended to minimize the thermal mismatch as much as possible. PMID- 18386845 TI - Electrospraying: a facile technique for synthesis of chitosan-based micro/nanospheres for drug delivery applications. AB - Electrospraying is a novel technique for the generation of micro/nanospheres for biomedical applications. Apart from being a high yield technique; electrospraying has an added advantage of not making use of an external dispersion/emulsion phase which often involves ingredients that are undesirable for biomedical applications. In this study, we report the use of electrospraying for the synthesis of chitosan micro/nanospheres. The focus was to optimize the fabrication parameters involved in electrospraying for reproducible synthesis of chitosan based micro/nanospheres and to study their potential as delivery vehicles for bioactive agents. The influence of the following was studied (i) electrospraying voltage, (ii) needle gauge, (iii) concentration of chitosan solution, (iv) concentration of acetic acid solution, and (v) electrospraying distance. SEM analysis demonstrated that microspheres of less than 1 mum were obtained when chitosan concentration was 2% dissolved in 90% acetic acid. The working distance and needle gauge that yielded favorable results were 7 cm and 26 g, respectively. Average particle size of ampicillin loaded chitosan micro/nanospheres was 520 nm with zeta potential of +28.2 mV and encapsulation efficiency of 80.4%. The particles were characterized for drug release kinetics and results demonstrated an initial burst release followed by a sustained release over a period of 120 h. Further, antibacterial activity of drug loaded micro/nanospheres demonstrated that the encapsulated drug was in its active form postexposure to high voltage during electrospraying. This study indicates that electrospraying is a facile technique for the synthesis of chitosan micro/nanospheres for drug delivery applications. PMID- 18386846 TI - Effect of bearing size on the long-term wear, wear debris, and ion levels of large diameter metal-on-metal hip replacements-An in vitro study. AB - Hip resurfacing arthroplasty has become a popular alternative to conventional hip surgery. Surface replacements with bearing sizes of 55 mm (n = 5) and 39 mm (n = 5) were tested in a hip simulator for 15 million cycles (Mc). Wear debris was isolated from the serum lubricant and characterized by field emmitting gun scanning electorn microscopy, and ion levels were measured via inductively coupled plasma mass spectroscopy at intervals throughout the test. The 39 mm bearings showed significantly greater bedding in volumetric wear (2.58 mm(3)) compared with the 55 mm bearings (1.15 mm(3)). There was no significant difference between the steady state wear rates (1-15 Mc) between the two sizes (0.10 and 0.09 mm(3)/Mc, respectively); however, this parity only became clear after 7 Mc. The wear debris isolated was oval in morphology with a mean particle size of 28 nm and a range of 9-108 nm. The Co levels measured at 0.13 Mc were significantly greater than at 3.6 Mc for both bearing sizes (10926 ppb and 176 ppb, respectively). After 0.5 Mc, the Co levels from the 39 mm bearings were significantly higher than the 55 mm (11,007 vs. 1475 ppb). The wear results support previous findings showing that increasing the femoral head size decreased volumetric bedding in wear. The ion levels measured suggest both bearing sizes have similar initial wear rates; however, the 55 mm bearings reach steady state wear more rapidly. PMID- 18386849 TI - Human cytomegalovirus load measurement and its applications for pre-emptive therapy in patients undergoing hematopoietic stem cell transplantation. AB - The hematopoietic stem cell transplantation (HSCT) setting is a rapidly evolving field, and procedures with a high degree of complexity are now entering into the clinic. The risk of developing opportunistic viral infections increases with the level of human leukocyte antigen disparity between the donor and the recipient of allogeneic HSCT and is enhanced by graft manipulations such as T-cell depletion or immune suppression required to control graft versus host disease (GVHD) or rejection. Among the opportunistic viral infections potentially hampering the outcome of HSCT, human cytomegalovirus (HCMV) infection plays a major role. In the early 1980s, before the introduction of effective antiviral treatment, HCMV disease accounted for the high mortality in recipients of allogeneic HSCT. In more recent years, the availability of HCMV-specific antiviral agents (ganciclovir, GCV, foscarnet, PFA and cidofovir, CDV) led to a dramatic decrease in HCMV-related morbidity and mortality. However, HCMV infection still remains a major concern in the HSCT setting. Historically, treatment of HCMV infection in HSCT recipients (HSCTR) evolved from treatment of overt HCMV disease to universal prophylaxis or pre-emptive treatment of active infection, in parallel with the improvement of techniques for diagnosing and monitoring HCMV infection in transplant recipients. PMID- 18386850 TI - Assessment of methoxyfenozide exposure on the sexual attractiveness and responsiveness of adult codling moth, Cydia pomonella L., in small orchard blocks. AB - BACKGROUND: The sexual attractiveness of female and the responsiveness of male codling moth, Cydia pomonella L., exposed to surfaces treated with the ecdysteroid agonist methoxyfenozide were investigated in small orchard block experiments. The main purpose of the study was to determine whether the reported sublethal effects of methoxyfenozide affecting moth behavior also occur under field conditions. RESULTS: Data from the first experiment showed that in some trials untreated females were significantly more attractive to wild males than methoxyfenozide-treated females. The second experiment revealed that some released methoxyfenozide-treated males were not as responsive to calling females as untreated males, and that some untreated females were more attractive to males than methoxyfenozide-treated females. The third experiment revealed that wild males exposed to treated tree surfaces were significantly less responsive to traps than were wild males exposed to untreated trees. However, there were no significant differences in mean recaptures of released males between the untreated and treated orchard blocks. CONCLUSION: Exposure of adult C. pomonella to methoxyfenozide-treated surfaces resulted in a negative impact on male responsiveness to calling females and synthetic pheromone lures. However, the level of impact on the sexual attractiveness of treated females did not appear to be as explicit. PMID- 18386851 TI - Sorption of sulfuryl fluoride by food commodities. AB - BACKGROUND: The use of sulfuryl fluoride, a structural fumigant for termite and woodborer control, has recently been expanded to treating stored food commodities and food facilities. There is, however, a lack of data on the sorption of sulfuryl fluoride by food commodities. Knowledge about sorption is important in the context of effective treatment and residues. RESULTS: When sulfuryl fluoride was applied at a dose of 50 g m(-3) to various food commodities (total 68) with 300 g per replicate in 0.75 L gas wash bottles (fumigation chambers) at 25 +/- 1 degrees C, in most cases (81%) the gas concentrations in the free space of the commodities exceeded 50 g m(-3) (range 51-80 g m(-3)) at the end of 24 h exposure. In chambers without the substrate, an average concentration of 49.7 g m(-3) was recorded. About 54% of the commodities showed low-level ( < or = 25%) sorption of sulfuryl fluoride, 34% showed medium-level (26-50%) sorption and only 12% were highly sorptive (>50%). The latter include white oats (terminal gas concentration 17.8 g m(-3)), some of the decorticated split pulses (24.0-29.3 g m(-3)), chickpea flour (26.3 g m(-3)), dried ginger (29.0 g m(-3)), refined wheat flour (30.3 g m(-3)) and coriander powder (40.5 g m(-3)). In unfumigated control commodities, owing to interfering volatiles, Fumiscope readings in the range 0-13 were noted. CONCLUSION: Sulfuryl fluoride has the advantage of a low or moderate level of sorption with the majority of the food commodities. PMID- 18386852 TI - Direct dynamics study of the hydrogen-abstraction reaction of 1,1,2,2,3 fluorinated propane with the hydroxyl radical. AB - The hydrogen abstraction reactions by a hydroxyl radical from 1,1,2,2,3 fluorinated propane (CF2HCF2CFH2) have been investigated by the dual-level direct dynamics method. Three equilibrium conformers (I, II, III) of CF2HCF2CFH2, one with Cs and two with C1 symmetries, are identified by the rotations of -CFH2 and CF2H groups. Two transition states are located for the conformer I (Cs symmetry) + OH --> products (R1) reaction, and three distinct transition states are identified for conformers II and III (C1 symmetry) + OH --> products (R2 and R3). The optimized geometries and harmonic vibrational frequencies of all reactants, complexes, transition states, and products are calculated at the BB1K/6-31+G(d,p) level of theory. The single-point energy calculations are performed at the G3(MP2) level using the BB1K geometries. Using improved canonical variational transition-state theory (ICVT) with the small-curvature tunneling correction (SCT), the rate constants for each channel are calculated over a wide temperature range of 200-2000 K. It is found that the H-abstraction reaction from the -CFH2 group is the predominant product channel for three reactions. The total rate constant is evaluated by the Boltzmann distribution function, and the agreement between theoretical and experimental values is good. PMID- 18386853 TI - Multiconfigurational g tensor calculations as a probe for the covalency of the copper-ligand bonds in copper(II) complexes: [CuCl4]2-, [Cu(NH3)4]2+, and plastocyanin. AB - Calculations of the g tensor of three copper(II) complexes [Cu(NH3)4]2+, [CuCl4]2 , and plastocyanin are presented. Two different sum-over-states-based approaches are considered, making use of the multistate CASPT2 method for excitation energies and PMCAS (perturbation modified CAS) wave functions for the computation of the angular momentum and spin-orbit coupling matrix elements. Test calculations on [Cu(NH3)4]2+ and [CuCl4]2- point to the need of including in the MS-CASPT2 treatment the specific charge-transfer state with an electron excited out of the bonding counterpart of the ground-state SOMO. The computed g shifts for these two molecules present a considerable improvement with respect to the results obtained from our previous g tensor calculations based instead on CASSCF/CASPT2. This is shown to be related to an improved description of the covalency of the Cu-L bonds. For the calculations on plastocyanin, different models are used, taken from a recent (QM/MM) DFT study by Sinnecker and Neese. The effect of the surrounding protein is taken into account by surrounding the central cluster either with a dielectric continuum (epsilon = 4) or with a set of point charges. The second approach is found to be indispensable for an accurate description of environmental effects. With this approach, the calculated g values compare to within 30 ppt with the experimental data of plastocyanin. PMID- 18386854 TI - Magnetic field effect on photoinduced electron transfer between [Cu(phen)2]2+ and DNA. AB - The magnetic field effect (MFE) on the photoinduced electron transfer (PET) reaction between the [Cu(phen)2]2+ complex and DNA has been studied in homogeneous buffer medium and in reverse micelles. The copper complex on photoexcitation can oxidize DNA in a deoxygenated environment. A prominent MFE is found even in a homogeneous aqueous medium for the triplet born radicals. The process of partial intercalation of [Cu(phen)2]2+ complex within DNA is responsible for such a rare observation. In reverse micelles, the MFE is not very much prominent because of the large separation distance between the component radicals of the geminate radical ion pairs generated through PET. PMID- 18386855 TI - Ultrafast charge-transfer-to-solvent dynamics of iodide in tetrahydrofuran. 2. Photoinduced electron transfer to counterions in solution. AB - The excited states of atomic anions in liquids are bound only by the polarization of the surrounding solvent. Thus, the electron-detachment process following excitation to one of these solvent-bound states, known as charge-transfer-to solvent (CTTS) states, provides a useful probe of solvent structure and dynamics. These transitions and subsequent relaxation dynamics also are influenced by other factors that alter the solution environment local to the CTTS anion, including the presence of cosolutes, cosolvents, and other ions. In this paper, we examine the ultrafast CTTS dynamics of iodide in liquid tetrahydrofuran (THF) with a particular focus on how the solvent dynamics and the CTTS electron-ejection process are altered in the presence of various counterions. In weakly polar solvents such as THF, iodide salts can be strongly ion-paired in solution; the steady-state UV-visible absorption spectroscopy of various iodide salts in liquid THF indicates that the degree of ion-pairing changes from strong to weak to none as the counterion is switched from Na+ to tetrabutylammonium (t-BA+) to crown ether-complexed Na+, respectively. In our ultrafast experiments, we have excited the I- CTTS transition of these various iodide salts at 263 nm and probed the dynamics of the CTTS-detached electrons throughout the visible and near-IR. In the previous paper of this series (Bragg, A. E.; Schwartz, B. J. J. Phys. Chem. B 2008, 112, 483-494), we found that for "counterion-free" I- (obtained by complexing Na+ with a crown ether) the CTTS electrons were ejected approximately 6 nm from their partner iodine atoms, the result of significant nonadiabatic coupling between the CTTS excited state and extended electronic states supported by the naturally existing solvent cavities in liquid THF, which also serve as pre existing electron traps. In contrast, for the highly ion-paired NaI/THF system, we find that approximately 90% of the CTTS electrons are "captured" by a nearby Na+ to form (Na+, e-)THF "tight-contact pairs" (TCPs), which are chemically and spectroscopically distinct from both solvated neutral sodium atoms and free solvated electrons. A simple kinetic model is able to reproduce the details of the electron capture process, with 63% of the electrons captured quickly in approximately 2.3 ps, 26% captured diffusively in approximately 63 ps, and the remaining 11% escaping out into the solution on subnanosecond time scales. We also find that the majority of the CTTS electrons are ejected to within 1 or 2 nm of the Na+. This demonstrates that the presence of the nearby cation biases the relocalization of CTTS-generated electrons from I- in THF, changing the nonadiabatic coupling to the extended, cavity-supported electronic states in THF to produce a much tighter distribution of electron-ejection distances. In the case of the more loosely ion-paired t-BA+-I-/THF system, we find that only 10-15% of the CTTS-ejected electrons associate with t-BA+ to form "loose-contact pairs" (LCPs), which are characterized by a much weaker interaction between the electron and cation than occurs in TCPs. The formation of (t-BA+, e-)THF LCPs is characterized by a Coulombically induced blue shift of the free eTHF- spectrum on a approximately 5-ps time scale. We argue that the weaker interaction between t BA+ and the parent I- results in little change to the CTTS-ejection process, so that only those electrons that happen to localize in the vicinity of t-BA+ are captured to form LCPs. Finally, we interpret the correlation between electron capture yield and counterion-induced perturbation of the I- CTTS transition as arising from changes in the distribution of ion-pair separations with cation identity, and we discuss our results in the context of relevant solution conductivity measurements. PMID- 18386856 TI - Solvation oscillations and excited-state dynamics of 2-amino- and 2-hydroxy-7 nitrofluorene and its 2'-deoxyriboside. AB - Push-pull substituted fluorenes are considered for use as dynamic solvation probes in polynucleotides. Their fluorescence band is predicted (by simulations) to show weak spectral oscillations on the subpicosecond time scale depending on the nucleotide sequence. The oscillations reflect the local far-infrared spectrum of the environment around the probe molecule. A connection is provided by the continuum theory of polar solvation which, however, neglects molecular aspects. We examine the latter using acetonitrile solution as a test case. A collective librational solvent mode at 100 cm(-1) is observed with 2-amino-7-nitrofluorene, 2-dimethylamino-7-nitrofluorene, 2-hydroxy-7-nitrofluorene, and its 2' deoxyriboside. Different strengths of the oscillation indicate that rotational friction of nearby acetonitrile molecules depends on the solute structure or that H bonding is involved in launching the librational coherence. Polar solvation in methanol is used for comparison. With hydroxynitrofluorenes, the observation window is limited by intersystem crossing for which rates are reported. A prominent excited-state absorption band of nitrofluorenes at 430 nm can be used to monitor polar solvation. Structural and electronic relaxation pathways are discussed with the help of quantum chemical calculations. PMID- 18386857 TI - Intersystem crossing mediated by photoinduced intramolecular charge transfer: julolidine-anthracene molecules with perpendicular pi systems. AB - Time-resolved electron paramagnetic resonance studies show that the primary mechanism of triplet formation following photoexcitation of julolidine-anthracene molecules linked by a single bond and having perpendicular pi systems is a spin orbit, charge-transfer intersystem crossing mechanism (SOCT-ISC). This mechanism depends on the degree of charge transfer from julolidine to anthracene, the dihedral angle (theta1) between their pi systems, and the magnitude of the electronic coupling between julolidine and anthracene. We compare 4-(9 anthracenyl)-julolidine with the more sterically encumbered 4-(9-anthracenyl)-3,5 dimethyljulolidine and find that fixing theta1 congruent with 90 degrees serves to enhance SOCT-ISC by increasing the change in orbital angular momentum accompanying charge transfer. Given that the requirements for the SOCT-ISC mechanism are quite general, we expect it to occur in a variety of electron donor acceptor systems. PMID- 18386859 TI - Quantum and molecular mechanical study of the first proton transfer in the catalytic cycle of cytochrome P450cam and its mutant D251N. AB - In the catalytic cycle of cytochrome P450cam, the hydroperoxo intermediate (Cpd 0) is formed by proton transfer from a reduced oxyheme complex (S5). This process is drastically slowed down when Asp251 is mutated to Asn (D251N). We report quantum mechanical/molecular mechanical (QM/MM) calculations that address this proton delivery in the doublet state through a hydrogen-bond network in the Asp251 channel, both for the wild-type enzyme and the D251N mutant, using four different active-site models. For the wild-type, we find a facile concerted mechanism for proton transfer from protonated Asp251 via Wat901 and Thr252 to the FeOO moiety, with a barrier of about 1 kcal/mol and a high exothermicity of more than 20 kcal/mol. In the D251N mutant with a neutral Asn251 residue, the proton transfer is almost thermoneutral or slightly exothermic in the three models considered. It is still very facile when the Asn251 residue adopts a conformation analogous to Asp251 in the wild-type enzyme, but the barrier increases significantly when the Asn251 side chain flips (as indicated by classical molecular dynamics simulations). This flip disrupts the hydrogen-bond network and hence the proton-transfer pathway, which causes a longer lifetime of S5 in the D251N mutant (consistent with experimental observations). The entry of an additional water molecule into the active site of D251N with flipped Asn251 regenerates the hydrogen-bond network and provides a viable mechanism for proton delivery in the mutant, with a moderate barrier of about 7 kcal/mol. PMID- 18386860 TI - Investigation on mechanism of catalysis by Pt-LiCoO2 for hydrolysis of sodium borohydride using X-ray absorption. AB - The synthesis of platinum nanoparticle loaded LiCoO2 (Pt-LiCoO2) was carried out successfully by an impregnation method followed by sintering at different temperatures. The catalytic role of Pt-LiCoO2 composite in hydrogen generation during hydrolysis of sodium borohydride (NaBH4) was studied for fuel cell applications. X-ray diffraction (XRD), transmission electron microscopy (TEM), and inductively coupled plasma-atomic emission spectroscopy (ICP-AES) have been used to elucidate the structural and catalytic properties of Pt-LiCoO2. It was found that the 15 wt % of Pt nanoparticles on LiCoO2 sintered at 450 degrees C support showed the maximum efficiency for the catalysis reaction of hydrogen production. X-ray absorption near edge structure (XANES) analysis and extended X ray absorption fine structure (EXAFS) analysis using a synchrotron radiation source were performed to carry out ex situ measurements in order to understand the mechanism of the catalytic process for the production of hydrogen during the hydrolysis of NaBH4. Co K-edge XANES showed a small percentage of cobalt in the metallic form after hydrogen generation which suggests the reduction of the cobalt during the hydrolysis of NaBH4. PMID- 18386861 TI - Combined X-ray diffraction and QM/MM study of the Burkholderia cepacia lipase catalyzed secondary alcohol esterification. AB - To understand the origin of high enantioselectivity of Burkholderia cepacia lipase (BCL) toward secondary alcohol, (R,S)-1-phenoxy-2-hydroxybutane (1), and its ester (E1), we determined the crystal structure of BCL complexed with phosphonate analogue of S-E1 and accomplished a series of MM, MC, and QM/MM studies. We have found that the inhibitor in the S configuration binds into the BCL active site in the same manner as the R isomer, with an important difference: while in case of the R-inhibitor the H-bond between its alcohol oxygen and catalytic His286 can be formed, in the case of the S-inhibitor this is not possible. Molecular modeling for both E1 enantiomers revealed orientations in which all hydrogen bonds characteristic of productive binding are formed. To check the possibility of chemical transformation, four different orientations of the substrate (two for each enantiomer) were chosen, and a series of ab initio QM/MM calculations were accomplished. Starting from the covalent complex, we modeled the ester (E1) hydrolysis and the alcohol (1) esterification. The calculations revealed that ester release is possible starting with all four covalent complexes. Alcohol release from the BCL-E1 complex in which the S substrate is bound in the same manner as the S-inhibitor in the crystal structure however is not possible. These results show that the crystallographically determined binding modes should be taken with caution when modeling chemical reactions. PMID- 18386862 TI - Optimized parameters for continuum solvation calculations with carbohydrates. AB - Continuum solvent models have been shown to be an efficient method for the calculation of the energetics of biomolecules in solution. However, for these methods to produce accurate results, an appropriate set of atomic radii or volumes is needed. While these have been developed for proteins and nucleic acids, the same is not true of carbohydrates. Here, a set of optimized parameters for continuum solvation calculations of carbohydrates in conjunction with the Carbohydrate Solution Force Field are presented. Explicit solvent free-energy perturbation simulations were performed on a set of hexapyranose sugars and used to fit atomic radii for Poisson-Boltzmann and generalized-Born calculations, and to fit atomic volumes for use with the analytical continuum electrostatics model. The solvation energetics computed with the optimized radii and a Poisson Boltzmann model show remarkable agreement with explicit solvent simulation, with a root-mean-square error of 1.19 kcal/mol over a large test set of sugars in many conformations. The generalized-Born model gives slightly poorer agreement, but still correlates very strongly, with an error of 1.69 kcal/mol. The analytical continuum electrostatics model correlates well with the explicit solvent results, but gives a larger error of 4.71 kcal/mol. The remarkable agreement between the solvation free energies computed in explicit and implicit solvent provides strong motivation for the use of implicit solvent models in the simulation of carbohydrate-containing systems. PMID- 18386863 TI - Temperature-dependent simultaneous ligand binding in human serum albumin. AB - Human serum albumin (HSA) is a soluble protein in our circulatory system, which is known to bind a variety of drugs and ligands. Since Sudlow's pioneering works on the ligand-binding sites, a major effort of the biophysical/biochemical research has been directed to characterize the structural, functional, and dynamical properties of this protein. Structural studies on HSA have revealed distinct temperature-induced folded states. Despite knowing about the ligand binding properties and residues important for the binding, less is understood about the temperature-dependent molecular recognition of the protein. Here, we have prepared thermally induced unfolded states of the protein and characterized those by circular dichroism (CD) and differential thermal analysis (DTA) techniques. The change in the globular structure of the protein as a consequence of thermal unfolding has also been characterized by dynamic light scattering (DLS) measurements. We have used two fluorescent ligands (4-(dicyanomethylene)-2 methyl-6-(p-dimethylaminostyryl) 4H-pyran) (DCM; hydrophobic; neutral) and Nile blue (NB; cationic) of different natures to characterize the ligand-binding properties of the protein in the native and thermally unfolded states. The possible binding sites of the ligands have been characterized by competitive binding with other drug molecules having definite binding sites in HSA. Picosecond-resolved Forster resonance energy transfer (FRET) studies along with steady-state and polarization-gated spectroscopies on the ligands in the protein reveal the dynamics of the binding sites at various temperatures. From the FRET studies, an attempt has been made to characterize the simultaneous binding of the two ligands in various temperature-dependent folded states of HSA. PMID- 18386865 TI - Highly ordered patterns of parabolic focal conics in lamellar lyotropic systems. AB - In this paper, we report the experimental observation of the formation of highly ordered parabolic focal conical patterns in lamellar surfactant solutions. Predominantly, we investigated mixtures of sodium dodecyl sulfate, water, hexanol, and decane, located in the immediate vicinity of the region of the L3 and Lalpha phase coexistence. The experimental studies of the formation of the patterns and of their temporal development are described. We give a simple model picture for the underlying structure, corroborated by the experimental results. There appears to be only one independent length scale that controls the appearance of the whole pattern. PMID- 18386864 TI - High-flux ceramic membranes with a nanomesh of metal oxide nanofibers. AB - Traditional ceramic separation membranes, which are fabricated by applying colloidal suspensions of metal hydroxides to porous supports, tend to suffer from pinholes and cracks that seriously affect their quality. Other intrinsic problems for these membranes include dramatic losses of flux when the pore sizes are reduced to enhance selectivity and dead-end pores that make no contribution to filtration. In this work, we propose a new strategy for addressing these problems by constructing a hierarchically structured separation layer on a porous substrate using large titanate nanofibers and smaller boehmite nanofibers. The nanofibers are able to divide large voids into smaller ones without forming dead end pores and with the minimum reduction of the total void volume. The separation layer of nanofibers has a porosity of over 70% of its volume, whereas the separation layer in conventional ceramic membranes has a porosity below 36% and inevitably includes dead-end pores that make no contribution to the flux. This radical change in membrane texture greatly enhances membrane performance. The resulting membranes were able to filter out 95.3% of 60-nm particles from a 0.01 wt % latex while maintaining a relatively high flux of between 800 and 1000 L/m2.h, under a low driving pressure (20 kPa). Such flow rates are orders of magnitude greater than those of conventional membranes with equal selectivity. Moreover, the flux was stable at approximately 800 L/m2.h with a selectivity of more than 95%, even after six repeated runs of filtration and calcination. Use of different supports, either porous glass or porous alumina, had no substantial effect on the performance of the membranes; thus, it is possible to construct the membranes from a variety of supports without compromising functionality. The Darcy equation satisfactorily describes the correlation between the filtration flux and the structural parameters of the new membranes. The assembly of nanofiber meshes to combine high flux with excellent selectivity is an exciting new direction in membrane fabrication. PMID- 18386866 TI - Effect of chirality on pi-stacking in styrene and maleimide alternating copolymers. AB - The effect of different chiral sequences in alternating copolymers of styrene and maleimide, poly(styrene-alt-dimethyl-N,N-propylamide) (SMI) was studied. The methods used for the SMI also applied to the general class of alternating copolymers of styrene and various maleimides. Only racemo-diisotactic polymers can associate into complexes because of the symmetrically distributed phenyl groups and maleimide monomers. This polymer is isotactic with respect to the chiral site of the styrene monomers (R, S) and isotactic with respect to the chirality of the maleimide [racemo (RR, SS)]. The maleimide, racemo (SS or RR) chirality, and the complementary S or R chirality in the styrene moiety form the repeating unit of racemo-diisotactic SMI. Structural deviations from the isotactic configurations induces an asymmetry in the distribution of styrene monomers and introduce bends in the polymer backbone, which prevent association through pi-stacking interactions. Semiempirical PM3 calculations on the SMI polymer have shown that this pi-stacking occurs when the polymers are separated by a distance about 10 A, and each pi-stacking pair formed from the styrene monomers has a stabilization energy of about 12 kJ/mol. PMID- 18386867 TI - UV-driven switching of chain orientation and liquid crystal alignment in nanoscale thin films of a novel polyimide bearing stilbene moieties in the backbone. AB - A novel photosensitive polyimide, poly(4,4'-stilbenylene 4,4'-oxidiphthalimide) (ODPA-Stilbene PSPI) was newly synthesized. The most surprising feature of this PSPI is that the PSPI films irradiated with linear polarized ultraviolet light (LPUVL) can favorably induce a unidirectional alignment of liquid crystals (LCs) in contact with the film surface and further switch the director of the unidirectionally aligned LCs from a perpendicular direction to a parallel direction with respect to the polarization direction of LPUVL by simply controlling the exposure dose in the irradiation process. These LPUVL-irradiated films were found to provide high anchoring energy to LCs, always giving very stable, homogeneous cells with unidirectionally aligned LCs regardless of the LC alignment directions. In the films, the PSPI polymer chains were found to undergo favorably unidirectional orientation via a specific orientation sequence of the polymer chain segments led by the directionally selective trans-cis photoisomerization of the stilbene chromophore units in the backbone induced by LPUVL exposure. Such unidirectionally oriented polymer chains of the films induce alignment of the LCs along the orientation direction of the polymer chains via favorable anisotropic molecular interactions between the oriented polymer chain segments and the LC molecules. In addition, the PSPI has an excellent film formation processibility; good quality PSPI thin films with a smooth surface are easily produced by simple spin-coating of the soluble poly(amic acid) precursor and subsequent thermal imidization process. In summary, this new PSPI is the promising LC alignment layer candidate with rubbing-free processing for the production of advanced LC display devices, including LC display televisions with large display areas. PMID- 18386868 TI - Nonlinear optical absorption properties of two multisubstituted p dimethylaminophenylethenyl pyridiniums. AB - Two multisubstituted D-pi-A type pyridinium salts, N-methyl-2,6-di[(E)-p dimethylaminophenylethenyl]pyridinium iodide (1) and N-methyl-2,4,6-tri[(E)-p dimethylaminophenylethenyl]pyridinium iodide (2), were synthesized with good yields by a modified method, and the structure of 1 was confirmed by X-ray diffraction. The nonlinear optical absorption (NOA) properties of the two compounds in the solution state were investigated by the Z-scan technique. Compounds 1 and 2 both exhibited very strong saturated absorption at picosecond pulse, and 1 displayed reverse saturated absorption, while 2 revealed saturated absorption properties at the nanosecond pulse. The electron-donating ability, number of side chains, and molecular coplanar characteristics of the compounds have a significant impact on their NOA properties. PMID- 18386869 TI - Unveiling the role of histidine and tyrosine residues on the conformation of the avian prion hexarepeat domain. AB - The prion protein (PrPC) is a glycoprotein that in mammals, differently from avians, can lead to prion diseases, by misfolding into a beta-sheet-rich pathogenic isoform (PrPSc). Mammal and avian proteins show different N-terminal tandem repeats: PHGGGWGQ and PHNPGY, both containing histidine, whereas tyrosine is included only in the primary sequence of the avian protein. Here, by means of potentiometric, circular dichroism (CD), and molecular dynamics (MD) studies at different pH values, we have investigated the conformation of the avian tetrahexarepeat (PHNPGY)4 (TetraHexaPY) with both N- and C-termini blocked by acetylation and amidation, respectively. We have found, also with the help of a recently proposed protein chirality indicator (Pietropaolo, A.; Muccioli, L.; Berardi, R.; Zannoni, C. Proteins 2008, 70, 667-677), a conformational dependence on the protonation states of histidine and tyrosine residues: the turn formation is pH driven, and at physiological pH a pivotal role is played by the tyrosine OH groups which give rise to a very compact bent structure of backbone upon forming a hydrogen-bond network. PMID- 18386870 TI - Preparation and characterization of new glassy system As2P2S8-Ga2S3. AB - Glasses having the composition (100 - x)As2P2S8-xGa2S3 with x ranging from 0 to 50% were investigated to determine the compositional effect on properties and local structure. The glass transition temperature (Tg) and the stability parameter against crystallization (Tx - Tg) increased with the addition of Ga2S3. The structure of these glasses was probed by Raman scattering, Fourier transform infrared (FT-IR) and 31P nuclear magnetic resonance. On the basis of the observed vibrations and the strength of the 31P-31P homonuclear magnetic dipolar coupling, two scenarios can be proposed for the structural evolution induced by the addition of Ga2S3. For x or= 30% we have depolymerization of the As2P2S8 units and the formation of a network of GaPS4 units with each PS 4/2 unit (Q4) species carrying a single positive formal charge. PMID- 18386871 TI - Dynamics of the self-assembling of mesogenic molecules in the prenematic region of isotropic liquid. AB - The linear and nonlinear dielectric relaxation spectra, recorded in the isotropic phase of mesogenic 4-(trans-4'-n-hexylcyclohexyl)isothiocyanatobenzene (6CHBT), reveal an equilibrium between the molecules that are involved and non-involved in the pseudo-nematic domains, which are spontaneously formed in the prenematic region. A perturbation of the equilibrium by a static electric field of high strength (E0 approximately 10(7) V/m) is followed by a nonlinear dielectric relaxation process that, measured with the probing electric field E(omega) of small amplitude and variable high frequency (up to 100 MHz), reflects the rate of the domains formation. The fraction of the mesogenic molecules involved in the prenematic domains at different temperatures is estimated. PMID- 18386872 TI - Hiking down the energy landscape: progress toward the Kauzmann temperature via vapor deposition. AB - Physical vapor deposition was employed to prepare amorphous samples of indomethacin and 1,3,5-(tris)naphthylbenzene. By depositing onto substrates held somewhat below the glass transition temperature and varying the deposition rate from 15 to 0.2 nm/s, glasses with low enthalpies and exceptional kinetic stability were prepared. Glasses with fictive temperatures that are as much as 40 K lower than those prepared by cooling the liquid can be made by vapor deposition. As compared to an ordinary glass, the most stable vapor-deposited samples moved about 40% toward the bottom of the potential energy landscape for amorphous materials. These results support the hypothesis that enhanced surface mobility allows stable glass formation by vapor deposition. A comparison of the enthalpy content of vapor-deposited glasses with aged glasses was used to evaluate the difference between bulk and surface dynamics for indomethacin; the dynamics in the top few nanometers of the glass are about 7 orders of magnitude faster than those in the bulk at Tg - 20 K. PMID- 18386873 TI - Dilute anionic surfactant solution route to polyaniline rectangular sub microtubes as a novel nanostructure. AB - A facile and low-cost approach has been developed for tailoring polyaniline rectangular sub-microtubes as a novel nanostructure of a conducting polymer in dilute sodium dodecyl sulfate (SDS) solution by the oxidation polymerization of aniline at room temperature. It was found that the size and uniformity of polyaniline rectangular sub-microtubes could be appropriately adjusted by tuning the concentration of aniline and the molar ratio of oxidant to aniline, respectively. The morphological evolution of rectangular sub-microtubes under different reaction times has been followed, and a possible formation mechanism has also been discussed in this report. The directing role of other anionic surfactants with -SO3(-) as the hydrophilic group for constructing polyaniline rectangular sub-microtubes has been investigated in detail. PMID- 18386874 TI - Anion binding in aqueous solutions by N-(Isonicotinamido)-N'-phenylthiourea-based simple synthetic neutral receptors. role of the hydrophobic microenvironment of the receptor molecule. AB - N-(Isonicotinamido)-N'-(substituted-phenyl)thioureas (1a-e, substituent X = p OCH3, p-CH3, H, m-Br, and m-CF3) have been designed as neutral receptors, in order to prove the influence of conformational issues on the ability to bind anions in aqueous solutions. Compounds 1a-e were shown to create a hydrophobic microenvironment around the thiourea group, favoring hydrogen bonding interactions, by evidence from quantum mechanic calculations, thermodynamic analysis, NMR aromatic current shielding, and comparative anion binding. Referring to N-(substituted-benzamido)thioureas (2a-e, substituent Y = H, m-Cl, m NO2, m,m-Cl,Cl, and p-NO2), we showed that, for the hydrophobic microenvironment to be operative in aqueous solutions, the amido -NH proton needs to be acidic enough. PMID- 18386875 TI - A density functional theory study of vibrational coupling in the amide I band of beta-sheet models. AB - We report the first molecular orbital/density functional theory (DFT) calculations on the vibrational frequencies involved in the amide I band of completely geometrically optimized models for beta-sheet peptides based upon (up to 16) glycine residues. These calculations use the B3LYP/D95** level of DFT. The primary means of vibrational coupling occurs through H bond, rather than through space, interactions, which is consistent with a previous report on alpha-helical polyalanines and H-bonding chains of both formamides and 4-pyridones. We decoupled the C=O stretching vibrations using selected 14C substitutions to probe the coupling mechanism and to determine "natural" frequencies for individual 14C=Os. The intermolecular H-bonding interactions affect the geometries of the amide groups. Those near the center of H-bonding chains have long C=O bonds. The C=O bond lengths correlate with these "natural" frequencies, The frequencies obtained from the DFT calculations are generally more coupled, and the most intense are more red shifted than those calculated by transition dipole coupling (TDC). TDC inverts the order of the shifted frequencies compared to DFT in several cases. PMID- 18386876 TI - Phenylene-ethynylene macrocycles as model systems of interchromophoric interactions in pi-conjugated macromolecules. AB - Unraveling the complex photophysics of macromolecular pi-conjugated systems requires both the development of suitable model systems to access a particular subset of a material's parameter space and the choice of matching spectroscopic techniques. We address the question of the strength of interchromophoric interactions in macromolecular systems by studying the fluorescence depolarization kinetics of a family of prototypical conjugated macrocycles. Shrinking the size of the molecular system decelerates fluorescence depolarization even though the radius of gyration decreases. Although the smaller macrocycles show faster rotational diffusion, the larger compounds exhibit an additional initial depolarization mechanism, attributed to intramolecular interchromophoric energy transfer. Comparison with fragments of the molecule illustrates that the larger macrocycles can be interpreted as bichromophoric systems, whereas the effectively parallel chromophoric elements of the smaller ring are indistinguishable in terms of polarization. The potential role of strong interchromophoric interaction is discussed. The results illustrate a subtle link between interchromophoric arrangement and ultrafast fluorescence depolarization, phenomena, which are often considered in the context of conjugated polymers: chromophoric alignment can potentially counteract the effect of polarization memory loss through energy transfer. PMID- 18386877 TI - A femtosecond study of excitation wavelength dependence of a triblock copolymer surfactant supramolecular assembly: (PEO)20-(PPO)70-(PEO)20 and CTAC. AB - Solvation dynamics and anisotropy decay of coumarin 480 (C480) in a supramolecular assembly containing a triblock copolymer, PEO20-PPO70-PEO20 (Pluronic P123) and a surfactant, CTAC (cetyl trimethylammonium chloride) are studied by femtosecond up-conversion. In a P123-CTAC complex, C480 displays a significant (22 nm) red edge excitation shift (REES) in the emission maximum as lambda ex increases from 335 to 445 nm. This suggests that the P123-CTAC aggregate is quite heterogeneous. The average rotational relaxation time (tau rot) of C480 in a P123-CTAC complex decreases by a factor of 2 from 2500 ps at lambda ex = 375 nm to 1200 ps at lambda ex = 435 nm. For lambda ex = 375 nm, the probe molecules in the buried core region of P123-CTAC are excited and the solvation dynamics displays three components, 2, 60, and 4000 ps. It is argued that insertion of CTAC in P123 micelle affects the polymer chain dynamics, and this leads to reduction of the 130 ps component of P123 micelle to 60 ps in P123 CTAC. For lambda ex = 435 nm, which selects the peripheral highly polar corona region, solvation dynamics in P123-CTAC and P123 are extremely fast with a major component of <0.3 ps ( approximately 80%) and a 2 ps ( approximately 20%) component. PMID- 18386878 TI - Relationship between the nonexponentiality of relaxation and relaxation time in the problem of glass transition. AB - By analyzing the experimental data for various glass-forming liquids and polymers, we find that the nonexponentiality, beta, and the relaxation time, tau, are commonly related: log(tau) is an approximately linear function of 1/beta, followed in most cases by a crossover to a higher linear slope. We rationalize this relationship in the recently developed elastic approach to the glass transition. The key to the observed common relationship between beta and tau is that the two quantities are governed by the same parameter, the liquid elasticity length, d el. The increase of d el on lowering temperature increases tau and decreases beta, resulting in the observed common relationship between beta and tau. In this picture, we also discuss the crossovers of beta and tau at low temperature. PMID- 18386879 TI - Simulations of surface forces in polyelectrolyte solutions. AB - We have simulated interactions between charged surfaces in the presence of oppositely charged polyelectrolytes by coupling perturbations in the isotension ensemble to a free energy variance minimization scheme. For polymeric systems, this method completely outperforms configurationally biased versions of grand canonical simulations. Proper diffusive equilibrium between bulk and slit has been established for polyelectrolytes with up to 60 monomers per chain. A consequence of imposing diffusive equilibrium conditions, in contrast to previous more restricted models, is the possibility of surface charge inversion; ion-ion correlation and the cooperativity of monomer adsorption drive the formation of a polyion layer close to the surface, that overcompensates the nominal surface charge. This is observed even at modest surface charge densities, and leads to a build up of a long ranged electrostatic barrier. In addition, the onset of charge inversion requires very low bulk polymer densities. Due to screening effects, this leads to a higher and more long-ranged free energy barrier at low, compared to high, bulk densities. Oscillatory forces, reminiscent of those found in simple hard sphere systems, are resolved in the high concentration regime. As a consequence of a second surface charge inversion, the system "stratifies" to form a stable polyelectrolyte layer in the central part of the slit, stabilized by the adsorbed surface layers. PMID- 18386880 TI - Intelligent gels and cryogels with entrapped emulsions. AB - Smart thermoresponsive gels and cryogels with incorporated emulsions have been synthesized and studied. The gels were obtained by three-dimensional copolymerization of N-isopropylacrylamide and N,N'-methylene-bis-acrylamide or N,N'-bis(acryloyl)cystamine in the presence of dispersion of tetradecane stabilized with sodium dodecylsulfate. Polymerization was performed at room temperature and below the water crystallization temperature. Both composite gels and cryogels were capable of heat-induced collapse. The extent of the collapse of the composite gel prepared at room temperature was much smaller and without squeezing of the lipophilic phase out of the shrunk composite gel. In contrast, shrinking of the composite cryogel was accompanied by release of tetradecane emulsion. PMID- 18386881 TI - Beta-homo-amino acid scan of angiotensin IV. AB - Angiotensin IV, a metabolite of angiotensin II, inhibits the enzyme insulin regulated aminopeptidase or IRAP and also, although with lower potency, aminopeptidase-N (AP-N). When both beta (2)-homo amino acid- and beta (3)-homo amino acid substitutions were used, allowed the identification of H-( R)beta (2)hVal-Tyr-Ile-His-Pro-beta (3)hPhe-OH as a potent and stable Ang IV analog with high selectivity for IRAP versus AP-N and the AT1 receptor. PMID- 18386882 TI - Catalytic cycles for the reduction of [UO2]2+ by cytochrome c7 proteins proposed from DFT calculations. AB - The mechanism of the reduction of the hydrated uranyl cation, [UO2](2+), by the cytochromes G. sulfurreducens and D. acetoxidans has been studied using density functional theory calculations. We propose that the initial electron transfer step from the heme is to a cation-cation complex in the case of D. acetoxidans, but for G. sulfurreducens, it is to a single uranyl cation, which then forms a U(V)-U(VI) complex with a second uranyl cation. For both enzymes, the subsequent catalytic pathways are very similar. A U(V)-U(V) complex is formed, which then undergoes disproportionation via two successive protonation steps of one uranyl group, to give a U(VI)-U(IV) complex which dissociates to individual U(VI) and U(IV) species, the former being bound at the enzyme active site. Intermediate structures along the catalytic pathway are consistent with EXAFS data. PMID- 18386883 TI - A combined metabolomic and proteomic investigation of the effects of a failure to express dystrophin in the mouse heart. AB - Muscle degeneration in the heart of 1-9 month-old mdx mice (a model for Duchenne muscular dystrophy) has been monitored using metabolomic and proteomic approaches. In both data sets, a pronounced aging trend was detected in control and mdx mice, and this trend was separate from the disease process. In addition, the characteristic increase in taurine associated with dystrophic tissue is correlated with proteins associated with oxidative phosphorylation and mitochondrial metabolism. PMID- 18386884 TI - 2-(6-aryl-3(Z)-hexen-1,5-diynyl)anilines as a new class of potent antitubulin agents. AB - Compounds 2a- h and 6 displayed significant GI 50 values of 10(-7)-10(-6) M against various cancer cell lines. Of these compounds, 2-(6-(2 trifluoromethylphenyl))-3(Z)-hexen-1,5-diynyl)aniline (2c) showed the most potent growth inhibition activity. Compound 2c also arrested cancer cells in the G2/M phase and in low concentration reduced a significant percentage of MDA-MB 231/ATCC breast cancer tetraploid cells. In addition to the G2/M block, compound 2c caused microtubule depolymerization and induced apoptosis via activation of the caspase family. PMID- 18386885 TI - Design and synthesis of peripherally restricted transient receptor potential vanilloid 1 (TRPV1) antagonists. AB - Transient receptor potential vanilloid 1 (TRPV1) channel antagonists may have clinical utility for the treatment of chronic nociceptive and neuropathic pain. We recently advanced a TRPV1 antagonist, 3 (AMG 517), into clinical trials as a new therapy for the treatment of pain. However, in addition to the desired analgesic effects, this TRPV1 antagonist significantly increased body core temperature following oral administration in rodents. Here, we report one of our approaches to eliminate or minimize the on-target hyperthermic effect observed with this and other TRPV1 antagonists. Through modifications of our clinical candidate, 3 a series of potent and peripherally restricted TRPV1 antagonists have been prepared. These analogues demonstrated on-target coverage in vivo but caused increases in body core temperature, suggesting that peripheral restriction was not sufficient to separate antagonism mediated antihyperalgesia from hyperthermia. Furthermore, these studies demonstrate that the site of action for TRPV1 blockade elicited hyperthermia is outside the blood-brain barrier. PMID- 18386886 TI - Spin-frustrated trinuclear Cu(II) clusters with mixing of 2(S = 1/2) and S = 3/2 states by antisymmetric exchange. 2. Orbital origin of in-plane Dzialoshinsky Moriya exchange parameters. AB - The microscopic origin of the in-plane (Gx, Gy) and out-of-plane (Gz) Dzialoshinsky-Moriya (DM) exchange parameters is considered for the Cu3(II) clusters. For the systems with the d(x2-y2) ground state of the Cu ions, only Z components of the pair DM exchange parameters are active (Gz not equal to 0, G(x,y) = 0) in the cases of the orientations of the local anisotropy axes zi| (zi||Z) and perpendicular (zi perpendicular Z, xi||(- Z)) to the molecular trigonal Z axis. The dependences of the Gx, Gy, and Gz DM exchange parameters on the tilt of the local magnetic orbitals were obtained for the antiferromagnetic (AFM) clusters with the d(x2-y2) and d(z2) ground state of the Cu ions. The tilt of the local d(x2-y2) orbitals results in the change of the Gz parameter and appearance of the in-plane DM exchange interactions (Gx or/and Gy parameters). The dependence of the Gz and Gx,Gy DM exchange parameters on the tilt angle is essentially different. The in-plane DM exchange coupling (Gx,Gy parameters) can significantly exceed the out-of-plane DM coupling (Gz parameter). The nonzero Gz and Gx,Gy parameters can be positive or negative. For the {Cu3} nanomagnet with the d(x2-y2) ground state and relatively strong DM coupling, the model explains the three DM exchange parameters of the same value (|Gz| = |Gx| = |Gy|) by the small tilt of the local anisotropy axes zi of the CuO4 local groups of the trimer from the positions zi perpendicular Z. The dependence of the DM exchange parameters (Gz, Gx, Gy) on the tilt for the AFM Cu3 clusters with the d(z2) ground states of the Cu ions differs significantly from that for the AFM systems with the ground state d(x2-y2) of the individual ions. Large in-plane DM exchange parameters Gx or/and Gy result in the mixing of the 2(S = 1/2) and S = 3/2 states and zero-field splitting (ZFS) 2D(DM) of the excited S = 3/2 state. The DM exchange contribution 2D(DM) to ZFS of the excited S = 3/2 state possesses the significant dependence on the tilt of the local magnetic orbitals. PMID- 18386887 TI - YF[MoO4] and YCl[MoO4]: two halide derivatives of yttrium ortho-oxomolybdate: syntheses, structures, and luminescence properties. AB - The halide derivatives of yttrium ortho-oxomolybdate YX[MoO 4] (X = F, Cl) both crystallize in the monoclinic system with four formula units per unit cell. YF[MoO 4] exhibits a primitive cell setting (space group P21/ c; a = 519.62(2) pm, b = 1225.14(7) pm, c = 663.30(3) pm, beta = 112.851(4) degrees ), whereas the lattice of YCl[MoO 4] shows face-centering (space group C2/m; a = 1019.02(5) pm, b = 720.67(4) pm, c = 681.50(3) pm, beta = 107.130(4) degrees ). The two compounds each contain crystallographically unique Y (3+) cations, which are found to have a coordination environment of six oxide and two halide anions. In the case of YF[MoO 4], the coordination environment is seen as square antiprisms, and for YCl[MoO 4], trigon-dodecahedra are found. The discrete tetrahedral [MoO 4] (2-) units of the fluoride derivative are exclusively bound by six terminal Y (3+) cations, while those of the chloride compound show a 5-fold coordination around the tetrahedra with one edge-bridging and four terminal Y (3+) cations. The halide anions in each compound exhibit a coordination number of two, building up isolated planar rhombus-shaped units according to [Y 2F 2] (4+) in YF[MoO 4] and [Y 2Cl 2] (4+) in YCl[MoO 4], respectively. Both compounds were synthesized at high temperatures using Y2O3, MoO3, and the corresponding yttrium trihalide in a molar ratio of 1:3:1. Single crystals of both are insensitive to moist air and are found to be coarse shaped and colorless with optical band gaps situated in the near UV around 3.78 eV for the fluoride and 3.82 eV for the chloride derivative. Furthermore, YF[MoO 4] seems to be a suitable material for doping to obtain luminescent materials because the Eu (3+)-doped compound shows an intense red luminescence, which has been spectroscopically investigated. PMID- 18386888 TI - Interpenetrating networks of three-dimensional Penrose tiles in CaAu3Ga, the structurally simplest cubic approximant of an icosahedral quasicrystal. AB - Double-Friauf polyhedra (DFPs) which play important roles in quasicrystal (QC) models are the unique building blocks in the novel 1/0 AC, CaAu(3+Delta)Ga(1 Delta) (Delta approximately equal to 0-0.13) [Pa_3; a = 9.0875(3)-9.1107(5) A]. The packing of DFPs generates interpenetrating networks of condensed three dimensional Penrose tiles, the geometry of which is close to that assumed for QCs. PMID- 18386889 TI - Lipid-packaged linear iron(II) triazole complexes in solution: controlled spin conversion via solvophobic self-assembly. AB - Linear Fe(II) 1,2,4-triazole complexes with lipid counteranions are newly developed. These complexes show sharp and reversible spin conversion in toluene, with temperatures significantly higher (by 20-100 K) than the spin crossover temperatures observed in the crystalline states. This is accounted for in terms of increased metal-ligand interactions in organic media, which is caused by solvophobic compaction of charged coordination chains. In atomic force microscopy, developed nanowires are observed for low spin (LS) complexes. On the other hand, fragmented nanostructures are seen for high spin (HS) complexes, indicating that the spin conversion in solution is governed by a self-assembly process. The lipid packaging of charged coordination chains thus provides powerful means to improve and regulate their functions via solvophobic self assembly. PMID- 18386890 TI - Morphology of large ZSM-5 crystals unraveled by fluorescence microscopy. AB - Understanding the internal structure of ZSM-5 crystallites is essential for improving catalyst performance. In this work, a combination of fluorescence microscopy, AFM, SEM, and optical observations is employed to study intergrowth phenomena and pore accessibility in a set of five ZSM-5 samples with different crystal morphologies. An amine-functionalized perylene dye is used to probe acid sites on the external crystal surface, while DAMPI (4-(4-diethylaminostyryl)- N methylpyridinium iodide) is used to map access to the straight channels in MFI from the outer surface. The use of these dyes is validated by studying the well understood rounded-boat type ZSM-5 crystals. Next coffin-shaped ZSM-5 crystals are considered; we critically evaluate the seemingly conflicting 2-component and 3-component models that have been proposed to account for the hourglass structure in these crystals. The data prove that observation of an hourglass structure is essentially unrelated to a 90 degree rotation of the pyramidal crystal components under the (010) face. Hence, in perfectly formed coffin-shaped crystals, the straight channels can be accessed from (010). However, in other crystal batches, sections with a 90 degrees rotation can be found; they are indeed located inside the crystal sections under (010) but often only partially occupy these pyramidal components. In such a case, both straight and sinusoidal pores surface at the hexagonal face. The results largely support the 3-component model, but with the added notion that 90 degree rotated sections (as proposed in the 2-component model) are most likely to be formed inside the defect-rich, pyramidal crystal sections under the (010) faces. PMID- 18386891 TI - Exceptional framework flexibility and sorption behavior of a multifunctional porous cuprous triazolate framework. AB - The porous metal azolate framework [Cu(etz)]infinity (MAF-2, Hetz = 3,5-diethyl 1,2,4-triazole) processes an NbO type cuprous triazolate scaffold and a CsCl type hydrophobic channel system, in which the large cavities are interconnected by small apertures with pendant ethyl groups. Since the ethyl-blocked apertures behave as thermoactivated IRIS stops for the guest molecules, the gas sorption behavior of MAF-2 can be controlled by temperature, in which N2 adsorption was observed at 195 K rather than 77 K. Single-crystal X-ray structural analysis revealed that the [Cu(etz)]infinity host framework is not altered upon N2 inclusion, confirming the occurrence of the so-called " kinetically controlled flexibility". By virtue of the kinetically controlled flexibility and hydrophobic pore surface, MAF-2 can adsorb large amounts of small organic molecules but excludes H2O. As demonstrated by single-crystal X-ray structural analyses, MAF-2 shrinks, expands, or distorts its framework to accommodate the hydrogen-bonded hexamers of MeOH, EtOH, or MeCN, respectively. Moreover, MAF-2 can also separate benzene and cyclohexane efficiently, as its flexible scaffold can distort to a certain degree so that benzene can diffuse through the distorted apertures but cyclohexane cannot. Moreover, the adsorption/desorption of these organic vapors induces reversible, multimode structural transformations. PMID- 18386892 TI - Spectral signatures and molecular origin of acid dissociation intermediates. AB - The existence of a broad, mid-infrared absorption ranging from 1000 to 3000 cm( 1) is usually interpreted as a signature for the existence of protonated water networks. Herein, we use cryogenic mixtures of water and hydrogen fluoride (HF) and show experimental and computational evidence that similarly wide absorptions can be generated by a broad distribution of proton-shared and ion pair complexes. In the present case, we demonstrate that the broadening is mainly inhomogeneous, reflecting the fact that the topology of the first solvation shell determines the local degree of ionization and the shared-proton asymmetric stretching frequency within H2O x HF complexes. The extreme sensitivity of the proton transfer potential energy hypersurface to local hydrogen bonding topologies modulates its vibrational frequency from 2800 down to approximately 1300 cm(-1), the latter value being characteristic of solvation geometries that yield similar condensed phase proton affinities for H2O and fluoride. By linking the local degree of ionization to the solvation pattern, we are able to propose a mechanism of ionization for HF in aqueous solutions and to explain some of their unusual properties at large concentrations. However, an important conclusion of broad scientific interest is our prediction that spectral signatures that are normally attributed to protonated water networks could also reveal the presence of strong hydrogen bonds between un-ionized acids and water molecules, with important consequences to spectroscopic investigations of biologically relevant proton channels and pumps. PMID- 18386893 TI - Necklace-like Cu@cross-linked poly(vinyl alcohol) core-shell microcables by hydrothermal process. AB - Unique necklace-like Cu@cross-linked poly(vinyl alcohol) (PVA) microcables have been discovered by a one-pot hydrothermal process. A new growth mechanism for such novel structures has been proposed. PMID- 18386894 TI - Characterization of Alzheimer's-like paired helical filaments from the core domain of tau protein using solid-state NMR spectroscopy. AB - The polymerization of the microtubule-associated protein tau into paired helical filaments (PHFs) represents one of the hallmarks of Alzheimer's disease. We employed solid-state nuclear magnetic resonance (NMR) to investigate the structure and dynamics of PHFs formed in vitro by the three-repeat-domain (K19) of protein tau, representing the core of Alzheimer PHFs. While N and C termini of tau monomers in PHFs are highly dynamic and solvent-exposed, the rigid segment consists of three major beta-strands. Combination of through-bond and through space ssNMR transfer methods with water-edited ((15)N, (13)C) and ((13)C, (13)C) correlation experiments suggests the existence of a fibril core that is largely built by repeat unit R3, flanked by surface-exposed units R1 and R4. Solid-state NMR, circular dichroism, and the fibrillization behavior of a K19 mutant furthermore indicate that electrostatic interactions play a central role in stabilizing the K19 PHFs. PMID- 18386895 TI - Rational design and gas-phase characterization of molecular capsules by self assembly of a symmetric hexasubstituted benzene with seven-membered lactams. AB - A novel very simple C3-symmetric lactam has been rationally designed to self assemble as dimers or larger platonic solid capsules. Its core flat benzene ring bears three seven-membered lactams, resulting in tripod molecules that aggregate into robust tetrameric capsules. The self-assembly process was templated by tetraethylammonium cations and proven to be reversible by ESI spectroscopy in various solvents. PMID- 18386896 TI - Asymmetric organocatalytic three-component 1,3-dipolar cycloaddition: control of stereochemistry via a chiral Bronsted acid activated dipole. AB - A Bronsted acid catalyzed three-component asymmetric 1,3-dipolar addition reaction between aldehydes, amino esters, and dipolarophiles by a new bisphosphoric acid, derived from the linked BINOL, furnished multiply substituted pyrrolidines in high yield with excellent enantioselectivities under mild conditions. PMID- 18386897 TI - A peptoid "antibody surrogate" that antagonizes VEGF receptor 2 activity. AB - We report a two-color, cell-based screen to identify specific receptor-binding compounds in a combinatorial library of peptoids displayed on beads. We apply this strategy to the isolation of vascular endothelial growth factor receptor 2 (VEGFR2)-binding peptoids. A dimeric derivative of one of these lead compounds is shown to be an antagonist of VEGFR2 activity both in vitro and in vivo. This methodology provides a potentially general route to synthetic molecules that bind integral membrane receptors with affinities and specificities similar to those of antibodies, but which are far smaller and easier to make and manipulate. PMID- 18386898 TI - De novo prediction of the structures of M. tuberculosis membrane proteins. AB - The structures of four integral membrane proteins from the Mycobacterium tuberculosis (TB) gene, Rv2433c, Rv1861, Rv1616, and Rv3069, have been de novo predicted by combining a generalized Born implicit solvent/membrane model with replica exchange molecular dynamics simulations to sample the conformational space of each protein. PMID- 18386899 TI - Extended X-ray absorption fine structure and nuclear resonance vibrational spectroscopy reveal that NifB-co, a FeMo-co precursor, comprises a 6Fe core with an interstitial light atom. AB - NifB-co, an Fe-S cluster produced by the enzyme NifB, is an intermediate on the biosynthetic pathway to the iron molybdenum cofactor (FeMo-co) of nitrogenase. We have used Fe K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy together with (57)Fe nuclear resonance vibrational spectroscopy (NRVS) to probe the structure of NifB-co while bound to the NifX protein from Azotobacter vinelandii. The spectra have been interpreted in part by comparison with data for the completed FeMo-co attached to the NafY carrier protein: the NafY:FeMo-co complex. EXAFS analysis of the NifX:NifB-co complex yields an average Fe-S distance of 2.26 A and average Fe-Fe distances of 2.66 and 3.74 A. Search profile analyses reveal the presence of a single Fe-X (X = C, N, or O) interaction at 2.04 A, compared to a 2.00 A Fe-X interaction found in the NafY:FeMo-co EXAFS. This suggests that the interstitial light atom (X) proposed to be present in FeMo co has already inserted at the NifB-co stage of biosynthesis. The NRVS exhibits strong bands from Fe-S stretching modes peaking around 270, 315, 385, and 408 cm( 1). Additional intensity at approximately 185-200 cm(-1) is interpreted as a set of cluster "breathing" modes similar to those seen for the FeMo-cofactor. The strength and location of these modes also suggest that the FeMo-co interstitial light atom seen in the crystal structure is already in place in NifB-co. Both the EXAFS and NRVS data for NifX:NifB-co are best simulated using a Fe 6S 9X trigonal prism structure analogous to the 6Fe core of FeMo-co, although a 7Fe structure made by capping one trigonal 3S terminus with Fe cannot be ruled out. The results are consistent with the conclusion that the interstitial light atom is already present at an early stage in FeMo-co biosynthesis prior to the incorporation of Mo and R-homocitrate. PMID- 18386901 TI - Influence of lipids in the generation of phenylacetaldehyde in wort-related model systems. AB - The effect of lipids on the formation of the Strecker aldehyde phenylacetaldehyde during wort boiling was studied to determine the role that small changes in the lipid content of the wort have in the production of significant flavor compounds in beer. Wort was treated with 0-2.77 mmol per liter of glucose, linoleic acid, or 2,4-decadienal and heated at 60-98 degrees C for 1 h. After this time, the amount of the Strecker aldehyde phenylacetaldehyde increased in the samples treated with linoleic acid or decadienal but not in the samples treated with glucose. Thus, the amount of phenylacetaldehyde produced in the presence of linoleic acid was 1.1-2.5 times the amount of the Strecker aldehyde produced in the control wort, and this amount increased to 3.6-4.6 times when decadienal was employed. The higher reactivity of decadienal than linoleic acid for this reaction decreased with temperature and was related to the oxidation of linoleic acid that occurred to a higher extent at higher temperatures. The above results suggest that lipids can contribute to the formation of Strecker aldehydes during wort boiling and that changes in the lipid content of the wort will produce significant changes in the formation of Strecker aldehydes in addition to other well-known consequences in beer quality and yeast metabolism. On the other hand, because of the high glucose content in wort, small changes in its content are not expected to affect the amount of Strecker aldehydes produced. PMID- 18386903 TI - Asymmetric nitroaldol reaction catalyzed by a C2-symmetric bisoxazolidine ligand. AB - A C2-symmetric bisoxazolidine was found to effectively catalyze the asymmetric Henry reaction of aliphatic and aromatic aldehydes. Beta-hydroxy nitroalkanes were produced in up to 99% yield and 95% ee. The bisoxazolidine-catalyzed nitroaldol formation requires relatively short reaction times, proceeds under mild conditions and is applicable to a wide range of substrates including sterically hindered aldehydes. PMID- 18386902 TI - Induction of apoptosis by vitamin D2, ergocalciferol, via reactive oxygen species generation, glutathione depletion, and caspase activation in human leukemia Cells. AB - This study demonstrated that ergocalciferol was able to inhibit leukemia cell growth in a concentration-dependent manner. Exploration of the acting mechanisms involved this event revealed that ergocalciferol induced DNA fragmentation and increased sub-G1 DNA contents in HL-60 cells, both of which are hallmarks of apoptosis. Analysis of the integrity of mitochondria demonstrated that ergocalciferol caused loss of mitochondrial membrane potential with release cytochrome c to cytosol, generation of reactive oxygen species (ROS), and depletion of glutathione (GSH), suggesting that ergocalciferol may induce apoptosis in HL-60 cells through a ROS-dependent pathway. Further results show that caspases-2, -3, -6, and -9 were all activated by ergocalciferol, together with cleavage of the downstream caspase-3 targets, DNA fragmentation factor (DFF 45), and poly(ADP-ribose) polymerase. In addition, ergocalciferol led to the increase in pro-apoptotic factor Bax accompanied with the decrease in anti apoptotic member Mcl-1, and the reduced Mcl-1 to Bax ratio may be a critical event concerning mitochondrial decay by ergocalciferol. Furthermore, ergocalciferol also led to induction of Fas death receptor closely linked to caspase-2 activation, suggesting the involvement of a Fas-mediated pathway in ergocalciferol-induced apoptosis. Totally, these findings suggest that ergocalciferol causes HL-60 apoptosis via a modulation of mitochondria involving ROS production, GSH depletion, caspase activation, and Fas induction. On the basis of anticancer activity of ergocalciferol, it may be feasible to develop chemopreventive agents from edible mushrooms or hop. PMID- 18386904 TI - Hexafluoroisopropanol as a unique solvent for stereoselective iododesilylation of vinylsilanes. AB - Stereoselective preparation of iodoalkenes from vinylsilanes is described. 1,1,1,3,3,3-hexafluoroisopropanol serves as a unique solvent that ensures high yields and stereoselectivities in the iododesilylation of a variety of functionalized substrates. PMID- 18386905 TI - Efficient intermolecular iron-catalyzed amidation of C-H bonds in the presence of N-bromosuccinimide. AB - We have developed an efficient, inexpensive, and air-stable catalyst/oxidant (FeCl2/NBS) system that could efficiently promote amidation of benzylic sp3 C-H bonds in ethyl acetate under mild conditions. PMID- 18386906 TI - Ir-catalyzed regio- and enantioselective Friedel-Crafts-type allylic alkylation of indoles. AB - Highly regio- and enantioselective Ir-catalyzed Friedel-Crafts type allylic alkylation of indoles have been realized using [Ir(COD)Cl]2/phosphoramidite ligand 1a, affording the branched products with up to >97/3 branched-linear ratio and 92% ee. PMID- 18386907 TI - Conformational domino effect in saccharides: a prediction from alkyl beta-(1-->6) diglucopyranosides. AB - A series of alkyl beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosides, containing nonchiral and chiral aglycons, were synthesized and analyzed by NMR and CD. The results, collected from four sets of disaccharides, demonstrated that the rotational properties of the interglycosidic linkage depend on the structural natures of both the aglycon and the solvent. Stereoelectronic and steric factors explain this rotational dependence, the gauche- trans (gt) rotamer being the most stable. Furthermore, correlations between Taft's steric parameters or between the pKa values of the alkyl substituent (aglycon) versus corresponding rotamer populations were observed. These results point to a natural conformational domino effect in oligosaccharides, where the conformational properties of each (1-->6) interglycosidic linkage will depend on the structure of the previous residue or its aglycon. In addition, a very weak rotational population dependence of the hydroxymethyl group at residue II on the aglycon at residue I was observed. The population of the gauche- gauche (gg) rotamer decreased, and that of gt increased as the Taft's steric parameters of the remote aglycon increased, independently of the disaccharide series and of the solvent. PMID- 18386908 TI - Palladium-catalyzed thiocarbonylation of iodoarenes with thiols in phosphonium salt ionic liquids. AB - Trihexyl(tetradecyl)phosphonium hexafluorophosphate, a phosphonium salt ionic liquid (PSIL) is a particularly effective general reaction media for the Pd catalyzed carbonylation reaction of iodoarenes and thiols to form thioesters. Recycling of the ionic liquid containing active Pd-catalyst was also demonstrated. PMID- 18386910 TI - On-chip electric field driven electrochemical detection using a poly(dimethylsiloxane) microchannel with gold microband electrodes. AB - An external electric field driven in-channel detection technique for on-chip electrochemical detection in micro fabricated devices is described based on a microfluidic system containing an array of 20 microband electrodes. It is shown that an external electric field induces a potential difference between two gold microband electrodes in a poly(dimethylsiloxane) (PDMS) microchannel, and that this enables the electrochemical detection of electroactive species such as ascorbic acid and Fe(CN) 6 (4-). The results, which are supported by simulations of the behavior of the microband electrodes in the microfluidic system, show that the induced potential difference between the electrodes can be controlled by altering the external electric field or by using different microbands in the microband array. As the obtained currents depend on the concentrations of electroactive species in the flowing solution and the detection can be carried out anywhere within the channel without interference of the external electric field, the present approach significantly facilitates electrochemical detection in capillary electrophoresis. This approach consequently holds great promise for application in inexpensive portable chip-based capillary electrophoresis (CE) devices. PMID- 18386909 TI - PCR-free quantitative detection of genetically modified organism from raw materials. An electrochemiluminescence-based bio bar code method. AB - A bio bar code assay based on oligonucleotide-modified gold nanoparticles (Au NPs) provides a PCR-free method for quantitative detection of nucleic acid targets. However, the current bio bar code assay requires lengthy experimental procedures including the preparation and release of bar code DNA probes from the target-nanoparticle complex and immobilization and hybridization of the probes for quantification. Herein, we report a novel PCR-free electrochemiluminescence (ECL)-based bio bar code assay for the quantitative detection of genetically modified organism (GMO) from raw materials. It consists of tris-(2,2'-bipyridyl) ruthenium (TBR)-labeled bar code DNA, nucleic acid hybridization using Au-NPs and biotin-labeled probes, and selective capture of the hybridization complex by streptavidin-coated paramagnetic beads. The detection of target DNA is realized by direct measurement of ECL emission of TBR. It can quantitatively detect target nucleic acids with high speed and sensitivity. This method can be used to quantitatively detect GMO fragments from real GMO products. PMID- 18386911 TI - Efficient optical trapping and visualization of silver nanoparticles. AB - We performed efficient optical trapping combined with sensitive optical detection of individual silver nanoparticles. The particles ranging in size from 20 to 275 nm in diameter were trapped in three dimensions using low laser power by minimizing spherical aberrations at the focus. The optical forces were quantified, and we found that the larger the particle, the stronger the optical force. The particles were imaged by an additional strongly scattered laser. PMID- 18386912 TI - Hierarchical agglomerates of carbon nanotubes as high-pressure cushions. AB - We report the cushioning behavior of highly agglomerated carbon nanotubes. The nanotube agglomerates can be repeatedly compacted to achieve large volume reduction (>50%) and expanded to nearly original volume without structural failure, like a robust porous cushion. At a higher pressure range (10-125 MPa), the energy absorbed per unit volume is 1 order of magnitude higher than conventional cushion materials such as foamy polystyrene. The structure of hierarchical agglomerates can be controlled for tailoring the cushioning properties and obtaining a lower cushioning coefficient (higher energy absorption) over a wide range of pressures (1-100 MPa). The mechanism was studied in terms of morphology evolution of the nanotube aggregates and pore size distribution during compression. PMID- 18386913 TI - Conductance of oriented C60 molecules. AB - C60 molecules adsorbed to Cu(100) are contacted with the tip of a cryogenic scanning tunneling microscope. Images with submolecular resolution reveal distinct orientations of the molecules. We find that the orientation significantly affects the conductance of the contact despite the high symmetry of C60. PMID- 18386914 TI - Screening-limited response of nanobiosensors. AB - Despite tremendous potential of highly sensitive electronic detection of biomolecules by nanoscale biosensors for genomics and proteomic applications, many aspects of experimentally observed sensor response (S) are difficult to understand within isolated theoretical frameworks of kinetic response or electrolyte screening. In this paper, we combine analytic solutions of Poisson Boltzmann and diffusion-capture equations to show that the electrostatic screening within an ionic environment limits the response of nanobiosensor such that S(t) approximately c1(ln(rho0) - ln(I0)/2 + ln(t)/ D F + c2[pH]) + c3 where c i are geometry-dependent constants, rho0 is the concentration of target molecules, I0 the salt concentration, and D F the fractal dimension of sensor surface. Our analysis provides a coherent theoretical interpretation of a wide variety of puzzling experimental data that have so far defied intuitive explanation. PMID- 18386915 TI - High-throughput, combinatorial engineering of initial codons for tunable expression of recombinant proteins. AB - We describe a high-throughput strategy for tuning the expression of recombinant proteins through engineering their early nucleotide sequences. After randomizing the +2 and +3 codons of the target genes, each of the variant genes was isolated in vivo and subsequently expressed using in vitro protein synthesis techniques. When several hundreds of clones were examined in parallel, it was found that expression levels of target genes varied as much as 70-fold depending on the identity of the codons in the randomized region. This broad and continuous distribution of expression levels enabled the selection of specific codon arrangements for the expression of target genes at a desired level. Furthermore, codon-dependent variations in protein expression were reproduced when the same genes were expressed in vivo. Thus, we expect that the methodology reported here could be utilized as a versatile platform for rapid expression of protein molecules at modulated levels either in vitro or in vivo. PMID- 18386916 TI - Chemical fragments as foundations for understanding target space and activity prediction. AB - The use of small inhibitors' fragment frequencies for understanding kinase potency and selectivity is described. By quantification of differences in the frequency of occurrence of fragments, similarities between small molecules and their targets can be determined. Naive Bayes models employing fragments provide highly interpretable and reliable means for predicting potency in individual kinases, as demonstrated in retrospective tests and prospective selections that were subsequently screened. Statistical corrections for prospective validation allowed us to accurately estimate success rates in the prospective experiment. Selectivity relationships between kinase targets are substantially explained by differences in the fragment composition of actives. By application of fragment similarities to the broader proteome, it is shown that targets related by sequence exhibit similar fragment preferences in small molecules. Of greater interest, certain targets unrelated by sequence are shown to have similar fragment preferences, even when the chemical similarity of ligands active at each target is low. PMID- 18386917 TI - Development of a novel membrane aerated hollow-fiber microbioreactor. AB - A new challenge in biotechnological processes is the development of flexible bioprocessing platforms, allowing strain selection, facilitating scale-up and integrating separation steps. Miniaturization of such a cultivation system allows parallel use and the saving of resources but makes the supply of oxygen to the cells difficult. In this work we present a membrane aerated hollow-fiber microbioreactor (HFMBR) which consists of an acrylic glass module equipped with two different types of membrane fibers. Fibers of polyethersulfone and polyvinyldifluoride were used for substrate and oxygen supply, respectively. Cultivation of E. coli as model organism and production of His-tagged GFP were carried out in the extracapillary space of the membrane aerated HFMBR and compared with cultivations in shaking flask which are commonly used for screening experiments. The measurement of the oxygen transfer capacity and the online monitoring of the dissolved oxygen during the cultivation were performed using a fiber optic oxygen sensor. Online measurement of the optical density was also integrated to the bioreactor. Due to efficient oxygen transfer, a better cell growth than in the shaking flask experiments was achieved, while no negative influence on the GFP productivity was observed in the membrane aerated bioreactor. Thus the feasibility of a future integrated downstreaming could also be demonstrated. PMID- 18386918 TI - Optimal economic design and operation of single- and multi-column chromatographic processes. AB - Single-column chromatography is widely used in the biopharmaceutical industries, although multi-column alternatives in the form of simulated moving bed (SMB) processes are now emerging. It may be difficult, however, to determine which column alternative will be best suited for a given application, and this work sets out to address this issue. A systematic approach is presented that is based on a full economic appraisal of each process alternative based on an optimization of the net annual profit. Single-column processes with and without recycling are considered, as are both the SMB and the Varicol process. The cyclic steady state for the SMB and Varicol processes is determined directly by complete discretization. The approach is applied to a case study based on a linear isotherm where it is found that for this particular system, a recycling policy is not necessary for the single column. When comparing the single-column process with the multi-column alternatives, the single column is the most economical provided the life time of the project is short; however, the economic benefits of the more capital-intensive multi-column processes are greater if the life time of the project is over 5 years. The SMB process is found to perform marginally better than the Varicol process over 15 years; however, this may be because not all extra degrees of freedom for the Varicol process were considered. PMID- 18386919 TI - Differences in the effects of solution additives on heat- and refolding-induced aggregation. AB - Although a number of low-molecular-weight additives have been developed to suppress protein aggregation, it is unclear whether these aggregation suppressors affect various aggregation processes in the same manner. In this study, we evaluated the differences in the effect of solution additives on heat- and refolding-induced aggregation in the presence of guanidine (Gdn), arginine (Arg), and spermidine (Spd), and the comparable analysis showed the following differences: (i) Gdn did not suppress thermal aggregation but increased the yield of oxidative refolding. (ii) Spd showed the highest effect for heat-induced aggregation suppression among tested compounds, although it promoted aggregation in oxidative refolding. (iii) Arg was effective for both aggregation processes. Lysozyme solubility assay and thermal unfolding experiment showed that Spd was preferentially excluded from native lysozyme and Arg and Gdn solubilized the model state of intermediates during oxidative refolding. This preference of additives to protein surfaces is the cause of the different effect on aggregation suppression. PMID- 18386920 TI - Determination by high-frequency and -field EPR of zero-field splitting in iron(IV) oxo complexes: implications for intermediates in nonheme iron enzymes. AB - [Fe(IV)O](2+) species have been implicated as the active form of many nonheme iron enzymes. The electronic structures of iron(IV) oxo complexes are thus of great interest. High-frequency and -field electron paramagnetic resonance is employed to determine accurately the spin Hamiltonian parameters of two stable complexes that contain the FeO unit: [FeO(TMC)(CH 3CN)](CF 3SO 3) 2, where TMC = tetramethylcyclam and [FeO(N4py)](CF 3SO 3) 2, where N4Py = bis(2 pyridylmethyl)bis(2-pyridyl)methylamine. Both complexes exhibit zero-field splittings that are positive, almost perfectly axial, and of very large magnitude: D = +26.95(5) and +22.05(5) cm (-1), respectively. These definitive experimental values can serve as the basis for further computational studies to unravel the electronic structures of such complexes. PMID- 18386921 TI - Structure and white luminescence of Eu-activated (Ba,Sr)(13-x)Al(22 2x)Si(10+2x)O66 materials. AB - The optical properties of Eu-activated (Ba,Sr)(13-x)Al(22-2x)Si(10+2x)O66 materials have been determined after the structural reinvestigation of the hypothetical Ba 13Al 22Si 10O 66 material on the basis of the Gebert's model. The white fluorescence and phosphorescence of the (Ba,Sr)(13-x)Al(22 2x)Si(10+2x)O66:Eu series result from the existence of two broad emission bands associated with (8)H-4f(6)5d(1)-->(8)S-4f(7) transitions peaking at 534 and 438 nm, the intensities of which may be tuned at room temperature via the control of the europium concentration and the substitution of Sr for Ba. This suggests the possibility to adjust the emission of the material to white LED requisites. PMID- 18386922 TI - The classic Wells-Dawson polyoxometalate, K6[alpha-P2W18O62].14H2O. Answering an 88 year-old question: what is its preferred, optimum synthesis? AB - The 88-year-old problem of developing a preferred, optimized synthesis of the prototype Wells-Dawson polyoxometalate, K6[alpha-P2W18O62].14H2O, is addressed herein. Specifically, six published syntheses of K6[alpha-P2W18O62].14H2O are listed and discussed, with emphasis given to the two most recent syntheses, Nadjo and co-workers' 2004 synthesis and a 1997 Inorganic Syntheses procedure by Droege, Randall, Finke et al. (hereafter D-R-F). For the starting experiment, the synthesis by Nadjo and co-workers was repeated. Next, the D-R-F synthesis and then the earlier (1984) synthesis in Droege's Ph.D. thesis were repeated and reinvestigated. The results demonstrate that the Nadjo synthesis produces over 200 g of high alpha-isomer purity (> or =97% by (31)P NMR) K6[alpha-P 2W18O62].14H2O in four steps over 8 days in 93% yield in our hands. A recrystallization step added as part of this work (for a total of five steps over 12 days) produces an increase in purity (>99%) with a concomitant loss of 8% yield (i.e., 85% overall yield) for the Nadjo-plus-recrystallization synthesis. Next, the D-R-F Inorganic Syntheses procedure was reinvestigated to determine the cause of "failed syntheses" occasionally encountered in our laboratories, the most recent and worst example to date being when one of us (C.R.G.) found 150 g of K10[alpha2-P 2W17O61] as an undesired side product when, as it turns out, the D-R-F Inorganic Syntheses procedure is followed rather than the earlier Droege synthesis. Specifically, it is shown that the problem in the Inorganic Syntheses procedure is that it ambiguously says to add 210 mL of HCl until a pH of 3-4 is reached when, in fact, it takes only 130-150 mL of HCL to reach a pH 3-4. Adding the full 210 mL of HCl ensures that a pH <2 is reached, as is required to produce isomerically pure K6[alpha-P 2W18O62].14H2O from the K 10[alpha 2-P 2W 17O 61] intermediate. The result is K6[alpha-P2W18O62].14H2O in five steps over 10 days in 82% yield and > or =97% purity. A table is provided comparing the details of the two best syntheses as reported herein: the Nadjo-plus-recrystallization synthesis and the D-R-F synthesis (with sufficient added HCl/proper pH control). That table makes apparent that the Nadjo-plus-recrystallization synthesis is improved on the basis of its better atom economy, its slightly higher product yields (85% vs 82%), slightly better purity (>99% vs >97%), and its comparable time (2 days shorter without recrystallization but 2 days longer with recrystallization) in comparison to the D-R-F synthesis with proper pH <2 control. Perhaps most importantly, some take-home messages concerning polyoxometalate synthesis illustrated by the iterative, 88 year-old quest to the best K6[alpha-P 2W18O62].14H2O synthesis are summarized and briefly discussed. PMID- 18386923 TI - Redox active cage for the electrochemical sensing of anions. AB - The tripodal system [1]3+ forms a 1:1 complex with CoII in which the metal is octahedrally coordinated by three bpy fragments. The [CoII(1)]5+ complex provides a cavity suitable for solvent or anion inclusion. X-ray diffraction studies on the crystalline complex salt of formula [CoII(1)...H2O]Cl(PF6)(4).2MeCN have shown that a water molecule is included in the cavity and the water oxygen atom receives six H-bonds from the C-H fragments of the three imidazolium subunits and of the three proximate pyridine rings, according to a slightly distorted trigonal prismatic geometry. Anion inclusion in an aqueous MeCN solution induces a distinct cathodic shift of the potential of the CoIII/CoII couple, whose magnitude decreases along the series: Cl->Br- approximately NCO->I- approximately NCS-, which reflects anion tendencies to receive H-bonds from the receptor. The variation of the water content in the MeCN solution (from 0 to 20%) induces a gradual change of the voltammetric response to anion titration: from two well distinguished peaks at a fixed potential to a single peak progressively shifted to a more cathodic potential. Such a behavior parallels the gradual decrease of the equilibrium constant for anion inclusion into the [CoII(1)]5+ receptor. PMID- 18386924 TI - One-step versus stepwise mechanism in protonated amino acid-promoted electron transfer reduction of a quinone by electron donors and two-electron reduction by a dihydronicotinamide adenine dinucleotide analogue. Interplay between electron transfer and hydrogen bonding. AB - Semiquinone radical anion of 1-(p-tolylsulfinyl)-2,5-benzoquinone (TolSQ(*-)) forms a strong hydrogen bond with protonated histidine (TolSQ(*-)/His x 2 H(+)), which was successfully detected by electron spin resonance. Strong hydrogen bonding between TolSQ(*-) and His x 2 H(+) results in acceleration of electron transfer (ET) from ferrocenes [R2Fc, R = C5H5, C5H4(n-Bu), C5H4Me] to TolSQ, when the one-electron reduction potential of TolSQ is largely shifted to the positive direction in the presence of His x 2 H(+). The rates of His x 2 H(+)-promoted ET from R2Fc to TolSQ exhibit deuterium kinetic isotope effects due to partial dissociation of the N-H bond in His x 2 H(+) at the transition state, when His x 2 H(+) is replaced by the deuterated compound (His x 2 D(+)-d6). The observed deuterium kinetic isotope effect (kH/kD) decreases continuously with increasing the driving force of ET to approach kH/kD = 1.0. On the other hand, His x 2 H(+) also promotes a hydride reduction of TolSQ by an NADH analogue, 9,10-dihydro-10 methylacridine (AcrH2). The hydride reduction proceeds via the one-step hydride transfer pathway. In such a case, a large deuterium kinetic isotope effect is observed in the rate of the hydride transfer, when AcrH2 is replaced by the dideuterated compound (AcrD2). In sharp contrast to this, no deuterium kinetic isotope effect is observed, when His x 2 H(+) is replaced by His x 2 D(+)-d6. On the other hand, direct protonation of TolSQ and 9,10-phenanthrenequinone (PQ) also results in efficient reductions of TolSQH(+) and PQH(+) by AcrH2, respectively. In this case, however, the hydride-transfer reactions occur via the ET pathway, that is, ET from AcrH2 to TolSQH(+) and PQH(+) occurs in preference to direct hydride transfer from AcrH2 to TolSQH(+) and PQH(+), respectively. The AcrH2(*+) produced by the ET oxidation of AcrH2 by TolSQH(+) and PQH(+) was directly detected by using a stopped-flow technique. PMID- 18386925 TI - Enantioselective organocatalytic Michael addition of aldehydes to nitroethylene: efficient access to gamma2-amino acids. AB - Enantioselective organocatalytic Michael addition of aldehydes to nitroethylene catalyzed by (S)-diphenylprolinol silyl ether provides beta-substituted-delta nitroalcohols in nearly optically pure form (96-99% ee). The Michael adducts bear a single substituent adjacent to the carbonyl and can be efficiently converted to protected gamma2-amino acids, which are essential for the systematic conformational studies of gamma-peptide foldamers. PMID- 18386926 TI - Low energy electron induced DNA damage: effects of terminal phosphate and base moieties on the distribution of damage. AB - Low energy electrons (LEE) induce DNA damage by dissociative electron attachment, which involves base release (N-glycosidic bond (N-C) cleavage) and the formation of strand breaks (phosphodiester-sugar bond (C-O) cleavage). The effect of terminal phosphate and base moieties was assessed by exposing DNA model compounds to LEE in the condensed phase followed by HPLC-UV analysis of products remaining on the surface. First, we report that the presence of terminal phosphate groups in monomers (pT, Tp, pTp) and dimers (pTpT, TpTp, pTpTp) increases overall damage by 2-3-fold while it decreases N-C and C-O bond cleavage by 2-10-fold. This suggests that the capture of LEE directly by the terminal phosphate does not contribute to N-C and C-O bond cleavage. Second, we report that terminal bases appear to shield the internal base from damage, resulting in a bias of damage toward the termini. In summary, the presence of terminal phosphate base moieties greatly affects the distribution of LEE induced damage in DNA model compounds. PMID- 18386927 TI - Peptide catalyzed asymmetric conjugate addition reactions of aldehydes to nitroethylene--a convenient entry into gamma2-amino acids. AB - The peptide H-D-Pro-Pro-Glu-NH2 is a highly effective catalyst for conjugate addition reactions between aldehydes and nitroethylene. Only 1 mol % of H-d-Pro Pro-Glu-NH2 and a 1.5-fold excess of aldehyde with respect to nitroethylene suffice to obtain gamma-nitroaldehydes and, after reduction, monosubstituted gamma-nitroalcohols in excellent yields and optical purities. The products can be readily converted into gamma2-amino acids, thereby opening an effective direct entry into this important class of compounds. PMID- 18386928 TI - Analysis of phenolic compounds by high-performance liquid chromatography and liquid chromatography/mass spectrometry in potato plant flowers, leaves, stems, and tubers and in home-processed potatoes. AB - Potato plants synthesize phenolic compounds as protection against bruising and injury from bacteria, fungi, viruses, and insects. Because antioxidative phenolic compounds are also reported to participate in enzymatic browning reactions and to exhibit health-promoting effects in humans, a need exists for accurate methods to measure their content in fresh and processed potatoes. To contribute to our knowledge about the levels of phenolic compounds in potatoes, we validated and used high-performance liquid chromatography and liquid chromatography/mass spectrometry to measure levels of chlorogenic acid, a chlorogenic isomer, and caffeic acid in flowers, leaves, stems, and tubers of the potato plant and in home-processed potatoes. The total phenolic acid content of flowers (626 mg/100 g fresh wt) was 21 and 59 times greater than that of leaves and stems, respectively. For all samples, chlorogenic acid and its isomer contributed 96-98% to the total. Total phenolic acid levels (in g/100 g fresh wt) of peels of five potato varieties grown in Korea ranged from 6.5 to 42.1 and of the flesh (pulp) from 0.5 to 16.5, with peel/pulp ratios ranging from 2.6 to 21.1. The total phenolic acid content for 25 American potatoes ranged from 1.0 to 172. The highest amounts were present in red and purple potatoes. Home processing of pulp with various forms of heat induced reductions in the phenolic content. The described methodology should facilitate future studies on the role of potato phenolic compounds in the plant and the diet. PMID- 18386929 TI - Analysis by HPLC and LC/MS of pungent piperamides in commercial black, white, green, and red whole and ground peppercorns. AB - Pepper plants accumulate pungent bioactive alkaloids called piperamides. To facilitate studies in this area, high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry methods were developed and used to measure the following piperamides in 10 commercial whole (peppercorns) and in 10 ground, black, white, green, and red peppers: piperanine, piperdardine, piperine, piperlonguminine, and piperettine. Structural identification of individual compounds in extracts was performed by associating the HPLC peak of each compound with the corresponding mass spectrum. The piperanine content of the peppers (in mg/g piperine equivalents) ranged from 0.3 for the ground white pepper to 1.4 in black peppercorns. The corresponding range for piperdardine was from 0.0 for seven samples to 1.8 in black peppercorns; for four isomeric piperines, from 0.7 for red to 129 in green peppercorns; for piperlonguminine, from 0.0 in red peppercorns to 1.0 in black peppercorns; and for piperyline, from 0.9 in ground black pepper to 5.9 for red peppercorn. Four well-separated stereoisomeric forms of piperettine with the same molecular weight were present in 19 peppers. The sums of the piperamides ranged from 6.6 for red to 153 for green peppercorns. In contrast to large differences in absolute concentrations among the peppers, the ratios of piperines to total piperamide were quite narrow, ranging from 0.76 for black to 0.90 for white peppercorns, with an average value of 0.84 +/- 0.04 ( n = 19). Thus, on average, the total piperamide content of the peppers consists of 84% piperines and 16% other piperamides. These results demonstrate the utility of the described extraction and analytical methods used to determine the wide ranging individual and total piperamide contents of widely consumed peppers. PMID- 18386931 TI - Enantioselective interactions at the solid-liquid interface of an HPLC column under working conditions. AB - A technique is presented which allows studying the enantioselective interactions occurring at the solid-liquid interface of a chiral stationary phase (CSP) and a racemate relevant to high performance liquid chromatography (HPLC). A conventional chiral column (Chiralpak AS) was mounted on an attenuated total reflection-infrared (ATR-IR) cell mimicking an HPLC setup equipped with an ATR-IR detector. Racemic pantolactone (PL) was used as the selectand. This setup in combination with modulation excitation spectroscopy (MES) allows for the identification of inter- and intramolecular hydrogen bonds being crucial for enantioseparation under HPLC operation conditions. The method is based on a two step strategy. In a first step, the enantiomers are separated by the chiral column similar to a standard HPLC experiment and upon adsorption on the identical CSP deposited on the internal reflection element (IRE), they are detected by ATR IR spectroscopy. This experiment provides a retention time for each enantiomer. From the difference in retention, a suitable frequency is calculated which is used in a second experiment where the racemate concentration is varied alternately (modulation) in a way that the pulses of ( R)-PL and ( S)-PL exhibit a phase lag of 90 degrees after elution through the column. This procedure allows one to gain separate information of the enantioselective selectand-CSP interaction after performing a demodulation similar to a phase sensitive detection (PSD). A further benefit of this method is the strong enhancement of the signal-to-noise ratio. The effectiveness of the method is demonstrated by investigating the observed faster decrease in retention time of the later-eluted ( R)-PL, as compared to ( S)-PL, when separating at higher temperatures (from 12 to 36 degrees C). The origin is attributed to a weakening of a specific hydrogen bond between the C=O of ( R)-PL and the N-H of the CSP. PMID- 18386930 TI - Differentiating proteomic biomarkers in breast cancer by laser capture microdissection and MALDI MS. AB - We assessed proteomic patterns in breast cancer using MALDI MS and laser capture microdissected cells. Protein and peptide expression in invasive mammary carcinoma versus normal mammary epithelium and estrogen-receptor positive versus estrogen-receptor negative tumors were compared. Biomarker candidates were identified by statistical analysis and classifiers were developed and validated in blinded test sets. Several of the m/ z features used in the classifiers were identified by LC-MS/MS and two were confirmed by immunohistochemistry. PMID- 18386932 TI - Solid-phase synthesis of aryl-substituted thienoindolizines: sequential Pictet Spengler, bromination and Suzuki cross-coupling reactions of thiophenes. AB - The solid-phase synthesis of a range of novel heterocyclic scaffolds based on the thiophene ring system, including thienoindolizines and aryl-substituted thiophenes, is presented. Specifically, a sequential methodology for the decoration of thienoindolizine scaffolds has been developed. This method involves a highly efficient and diastereoselective intramolecular Pictet-Spengler reaction, a quantitative and regioselective bromination of the thiophene ring, and a final Suzuki cross-coupling with an arylboronic acid. Crude products were generally obtained in high purities (>90%). In addition, an investigation on the acidic and electronic effects governing the rate of the Pictet-Spengler reactions was performed. Finally, a range of substituted thiophenes was attached to solid supports and subjected to the regioselective bromination and Suzuki cross coupling reactions, thus providing substituted thiophenes with high purities of crude products. PMID- 18386934 TI - Nanowire lithography on silicon. AB - Nanowire lithography (NWL) uses nanowires (NWs), grown and assembled by chemical methods, as etch masks to transfer their one-dimensional morphology to an underlying substrate. Here, we show that SiO2 NWs are a simple and compatible system to implement NWL on crystalline silicon and fabricate a wide range of architectures and devices. Planar field-effect transistors made of a single SOI NW channel exhibit a contact resistance below 20 kOmega and scale with the channel width. Further, we assess the electrical response of NW networks obtained using a mask of SiO2 NWs ink-jetted from solution. The resulting conformal network etched into the underlying wafer is monolithic, with single-crystalline bulk junctions; thus no difference in conductivity is seen between a direct NW bridge and a percolating network. We also extend the potential of NWL into the third dimension, by using a periodic undercutting that produces an array of vertically stacked NWs from a single NW mask. PMID- 18386933 TI - Real-time intravital imaging of RGD-quantum dot binding to luminal endothelium in mouse tumor neovasculature. AB - Nanoscale materials have increasingly become subject to intense investigation for use in cancer diagnosis and therapy. However, there is a fundamental dearth in cellular-level understanding of how nanoparticles interact within the tumor environment in living subjects. Adopting quantum dots (qdots) for their excellent brightness, photostability, monodispersity, and fluorescent yield, we link arginine-glycine-aspartic acid (RGD) peptides to target qdots specifically to newly formed/forming blood vessels expressing alpha vbeta 3 integrins. Using this model nanoparticle system, we exploit intravital microscopy with subcellular ( approximately 0.5 microm) resolution to directly observe and record, for the first time, the binding of nanoparticle conjugates to tumor blood vessels in living subjects. This generalizable method enabled us to show that in this model qdots do not extravasate and, unexpectedly, that they only bind as aggregates rather than individually. This level of understanding is critical on the path toward ensuring regulatory approval of nanoparticles in humans for disease diagnostics and therapeutics. Equally vital, the work provides a platform by which to design and optimize molecularly targeted nanoparticles including quantum dots for applications in living subjects. PMID- 18386935 TI - A lossless negative dielectric constant from quantum dot exciton polaritons. AB - Prospects for a lossless negative dielectric constant material for optical devices are studied. Simulations show that with sufficient gain, a mixture of two semiconductor quantum dots (QDs) can produce an effective dielectric constant that is lossless and negative. This permits, in concept, arbitrarily small scaling of the optical mode volume, a major goal in the field of nanophotonics. The proposed implementation of a lossless negative dielectric constant material based on colloidal QDs opens a tractable path. PMID- 18386936 TI - Spatiotemporal response of living cell structures in Dictyostelium discoideum with semiconductor quantum dots. AB - The ability to monitor the spatial and temporal organization of molecules such as biopolymers within a cell is essential to enable the ability to understand the complexity and dynamics existing in biological processes. However, many limitations currently exist in specifically labeling proteins in living cells. In our study, we incorporate nanometer-sized semiconductor quantum dots (QDs) into living cells for spatiotemporal protein imaging of actin polymers in Dictyostelium discoideum without the necessity of using complicating transmembrane transport approaches. We first demonstrate cytoplasmic distribution of QDs within these living amoebae cells and then show molecular targeting through actin filament labeling. Also, we have developed a microfluidic system to control and visualize the spatiotemporal response of the cellular environment during cell motility, which allows us to demonstrate specific localization control of the QD-protein complexes in living cells. This study provides a valuable tool for the specific targeting and analysis of proteins within Dictyostelium without the encumbrance of transmembrane assisted methods, which has implication in fields including polymer physics, material science, engineering, and biology. PMID- 18386937 TI - Controlled growth of ternary alloy nanowires using metalorganic chemical vapor deposition. AB - We report the growth and characterization of ternary AlxGa1- xAs nanowires by metalorganic chemical vapor deposition as a function of temperature and V/III ratio. Transmission electron microscopy and energy dispersive X-ray spectroscopy show that, at high temperatures and high V/III ratios, the nanowires form a core shell structure with higher Al composition in the nanowire core than in the shell. We develop a growth model that takes into account diffusion of reactants and decomposition rates at the nanowire catalyst and stem to describe the compositional difference and the shell growth rate. Utilizing this model, we have successfully grown compositionally uniform Al0.16Ga0.84As nanowires. The ability to rationally tune the composition of ternary alloy nanowires broadens the application range of nanowires by enabling more complex nanowire heterostructures. PMID- 18386938 TI - Quantifying metabolic activity of filamentous fungi using a colorimetric XTT assay. AB - The filamentous nature and robust cell walls of many fungi render traditional measurements of active biomass (e.g., turbidity, dry cell weight) ineffective for most fungal bioprocesses. To overcome this challenge, an assay for quantification of overall metabolic activity is developed using 2,3-bis(2-methoxy-4-nitro-5 sulfophenly)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT), which in the presence of active mitochondria is converted to a water-soluble formazan derivative that absorbs light in the visible spectrum (430-490 nm). Tests on the model fungus Aspergillus nidulans show that in actively growing cultures XTT absorbance is linearly related to dry cell weight below 0.2 g/kg broth. Validation through growth rate testing shows the developed XTT assay is able to accurately quantify reductions in culture metabolism during damaging physical treatment (heat, high shear, microwaving). Experiments in batch culture demonstrate that the developed XTT assay is capable of reporting on metabolic activity where dry cell weight is not. The developed assay is inexpensive, relatively rapid, and easy to conduct, making it ideally suited for assessment of fungal processes in the biotechnology industry. PMID- 18386939 TI - Process monitoring the inactivation of ricin and model proteins by disinfectants using fluorescence and biological activity. AB - It is important to develop rapid and reliable processes to monitor the decontamination of toxins released to the environment. The inactivation of the protein toxin ricin by the disinfectants bleach (sodium hypochlorite) and monochloramine was measured by the effect on mammalian cell cytotoxicity. The effect of the disinfectants on the native fluorescence (due mainly to tryptophan and to a lesser extent tyrosine) of ricin was also measured in parallel. Reactions of the disinfectants resulted in a decrease in the native fluorescence that was measured in real time in a noninvasive manner. We compared the inactivation of two well-characterized model enzymes to the behavior of ricin. The model enzymes studied were lysozyme, a small basic enzyme stabilized with internal disulfide bonds, and heart-muscle-type lactate dehydrogenase (LDH), a large protein composed of four subunits. The biological activities of the model enzymes were measured in parallel with their fluorescence. Gel electrophoresis showed a large number of modifications of the proteins caused by the disinfectants reflected in changes in mobility and the formation of higher-order aggregates. Size-exclusion chromatography showed that the disinfectants did not break down the subunit structure of ricin but instead resulted in an increased size and heterogeneity of the protein. Size-exclusion chromatography of LDH indicated that the subunits were dissociated and that higher-order aggregates were also formed. Bleach caused a rapid inactivation of biological activity correlated with a rapid decrease in the fluorescence. Monochloramine required much higher concentrations for significant effects and the kinetics of the reactions were slow, with half-life values of the decrease on the order of minutes. Each protein showed individual differences in responses to the disinfectants, but there was a consistent correlation between the loss of fluorescence and the decrease in biological activity. These results indicate that the monitoring the fluorescence is a useful process with limitations that can be used to monitor the inactivation of toxins using disinfectants. PMID- 18386940 TI - Pressure-induced phase transitions on a liquid crystalline europium(III) complex. AB - The effect of pressure on the phase behavior of the liquid crystalline complex [Eu(bta)(3)L(2)] (bta is benzoyltrifluoroacetonate, and L is the Schiff base 2 hydroxy-N-octadecyl-4-tetradecyloxybenzaldimine) was studied by X-ray diffraction, Raman spectroscopy, and luminescence spectroscopy. The pressure was varied between ambient pressure and 8.0 GPa. [Eu(bta)(3)L(2)] exhibits a smectic A (SmA) phase at room temperature. The complex undergoes a transition from the SmA phase to a solid lamellar structure around 0.22 GPa and another transition from the solid lamellar phase to an amorphous state from 1.6 to 3.5 GPa. At low pressures, the smectic layer spacing increases, and the intermolecular distance decreases. Above 3.5 GPa, both the interlamellar and the intermolecular spacings hardly change, but the intensity of X-ray reflections exhibits a remarkable decrease and eventually vanishes. An interpretation of the changes in the molecular structure is given. It was found that less interdigitation of the alkyl chains situated in adjacent layers and/or a full extension of the alkyl chains occurred at low pressures and that the second phase transition was accompanied by a transfer of the hydrogen atom from the nitrogen atom of the imine group to the oxygen atom of the Schiff base ligand. The effect of applying pressure equals that of the lanthanide contraction on the phase behavior. PMID- 18386941 TI - Mesophase separation in polyelectrolyte-mixed micelle coacervates. AB - Mesophase separation has been identified in a polycation/anionic-nonionic mixed micelle system formed by the coacervation of poly(diallyldimethylammoniumchloride)/sodium dodecylsulfate-Triton X-100 in 0.40 M NaCl. The resultant dense, optically clear fluid was studied by turbidity, dynamic light scattering (DLS), and rheology. The presence of two diffusion modes in DLS points to microscopic heterogeneity: coexistence of micelle-rich (dense) domains with micelle-poor (dilute) domains. With an increase in temperature above 20 degrees C, the turbidity rises rapidly along with the intensity of the slow mode. The concomitant decrease in the diffusivity of the slow mode signals an increase in the effective viscosity of the dense domain. With further increase in temperature, dramatic shear thinning is observed, and finally, macroscopic phase separation can be identified by centrifugation. At a temperature near that for quiescent phase separation, we observe shear-induced phase separation. We propose a mechanism to explain the connection between temperature- and shear-induced mesophase separation. PMID- 18386942 TI - Effects of flame treatment on the interfacial energy of polyethylene assessed by contact mechanics. AB - The effects of flame treatment of low-density polyethylene (LDPE) on the work of adhesion (W) and energy release rate (G) were assessed by a custom-built adhesion testing device (ATD). The contact area and the vertical displacement between planar LDPE films and PDMS lenses (untreated and UV/ozone treated) were varied as a function of the applied load, P. The adhesion and pull-off forces between the extracted PDMS lenses and the LDPE films were studied as a function of the duration of the flame treatment expressed in treatment numbers. A fracture mechanics approach was used to relate the applied load and the radius of the contact area to the energy release rate (G). The difference between the energy release rate and the thermodynamic work of adhesion, defined as the adhesion hysteresis (G - W), increased systematically with increasing treatment numbers for both lens types. In addition, the adhesion hysteresis appeared to be dependent on the contact time. Finally, the origin of the adhesion hysteresis was discussed. It was concluded that bonding by surface functional groups may be the dominating mechanisms for the changes in the observed hysteresis. PMID- 18386943 TI - Completely dispersible PEGylated gold nanoparticles under physiological conditions: modification of gold nanoparticles with precisely controlled PEG-b polyamine. AB - A novel water-soluble, biocompatible polymer, poly(ethylene glycol)-block-poly((2 N,N-dimethylamino)ethyl methacrylate) (PEG-b-PAMA), possessing controlled molecular weight with a narrow molecular weight distribution, was synthesized by the atom-transfer radical polymerization (ATRP) method. PEG-b-PAMA having a short PAMA chain length was successfully synthesized under suitable polymerization conditions. Gold nanoparticles (GNPs) were modified using PEG-b-PAMA prepared under a variety of PEGylation conditions. Under alkaline conditions (pH >10) and an [N]/[GNP] ratio of more than 3300, the PEGylated GNPs (PEG-GNPs) showed complete dispersion stability, avoiding coagulation. The amino groups of the PAMA segment of the block copolymers were completely deprotonated above pH 10. This means that PEG-b-PAMA interacted with the GNP surface via multipoint coordination of the tertiary amino groups of PAMA, not electrostatically. The effect of the number of amino groups in the PAMA segment on GNP surface modifications was investigated by zeta potential and dynamic light scattering (DLS) measurements. When the PEG-GNPs were prepared in excess polymer solution, almost the same diameter was observed regardless of the PAMA chain length. After the PEG-GNPs were purified by centrifugation, the zeta potentials of all PEG-GNPs were shielded to almost 0 mV, indicating the effective modifications of the GNP surface by PEG-b-PAMA regardless of the chain length. However, the particle size and particle size distribution of the purified PEG-GNPs were strongly affected by the PAMA chain length. PEG-GNPs with longer PAMA segments underwent coagulation after purification, whereas PEG-GNPs with shorter PAMA segments increased their dispersion stability. The experimental results of the thermal gravimetric analysis confirmed that the PEG density on the GNP surface increased as the AMA units decreased to 3. Thus, the dispersion stability depended significantly on the PEG density on the GNP surface. GNPs modified with PEG-b-PAMA having short AMA units showed excellent dispersion stability under a variety of pH conditions. The excellent dispersion stability of the obtained PEG-GNP was also confirmed both in bovine serum albumin (BSA) solution and 95% human serum. PMID- 18386944 TI - Cadmium, lead, and mercury levels in feathers of small passerine birds: noninvasive sampling strategy. AB - Bird feathers have been widely used as a nondestructive biological material for monitoring heavy metals. Sources of metals taken up by feathers include diet (metals are incorporated during feather formation), preening, and direct contact with metals in water, air, dust, and plants. In the literature, data regarding the origin of trace elements in feathers are not univocal. Only in the vast literature concerning mercury (as methyl mercury) has endogenous origin been determined. In the present study, we investigate cadmium, lead, and mercury levels in feathers of prey of Falco eleonorae in relation to the ecological characteristics (molt, habitat, and contamination by soil) of the different species. Cluster analysis identified two main groups of species. Differences and correlations within and between groups identified by cluster analysis were then checked by nonparametric statistical analysis. The results showed that mercury levels had a pattern significantly different from those of cadmium and lead, which in turn showed a significant positive correlation, suggesting different origins. Nests of F. eleonorae proved to be a good source for feathers of small trans-Saharan passerines collected by a noninvasive method. They provided abundant feathers of the various species in a relatively small area--in this case, the falcon colony on the Isle of San Pietro, Sardinia, Italy. PMID- 18386945 TI - Enhanced esterase activity and resistance to azinphosmethyl in target and nontarget organisms. AB - The organophosphorous compound azinphosmethyl (AzMe) is applied extensively in northern Patagonia (southern Argentina) to manage codling moths (Cydia pomonella). This area is irrigated by fast-flowing channels that provide a favorable habitat for many species, including amphipods (Hyalella curvispina) and a field-mixed population of black flies (Simulium bonaerense, Simulium wolffhuegeli, and Simulium nigristrigatum). In the present study, AzMe susceptibility and carboxylesterase (CarbE) activity from both insecticide exposed and nonexposed field populations were studied. The median lethal dose determined in codling moths from an insecticide-treated orchard was significantly higher (3.48 microg/insect) than that observed in those from an untreated orchard (0.69 microg/insect). Similarly, the median lethal concentration (LC50) determined in black flies collected from the treated area (0.021 mg/L) was significantly higher than that recorded in those from the untreated site (0.011 mg/L). For amphipods, both a subpopulation susceptible to AzMe (LC50, 1.83 microg/L) and a resistant one (LC50, 390 microg/L) were found in the treated area. Both subpopulations were more resistant to AzMe than the population from the untreated site (LC50, 0.43 microg/L). Significant differences (p < 0.001) in CarbE activities were observed between populations from pesticide-treated and untreated areas. Mean activities +/- standard deviation from treated and untreated sites were 0.21 +/- 0.16 and 0.016 +/- 0.008 micromol/min/mg protein, respectively, for codling moths; 2.17 +/- 1.71 and 0.81 +/- 0.35 micromol/min/mg protein, respectively, for black flies; and 0.27 +/- 0.10 and 0.14 +/- 0.07 micromol/min/mg protein, respectively, for amphipods. The results suggest that enhanced CarbE activity is one of the mechanisms that provide AzMe resistance in H. curvispina, Simulium spp., and C. pomonella populations from the insecticide treated areas. PMID- 18386948 TI - Advances in contemporary palliative and supportive care. PMID- 18386949 TI - Back to our roots: palliative care in the clinical setting. PMID- 18386950 TI - Narratives of a man with severe chronic heart failure and his wife in palliative advanced home care over a 4.5-year period. AB - The aim of this paper is to illuminate meanings of living the 'ups and downs', through the narratives of one man with severe, chronic heart failure (CHF) and his wife in palliative advanced home care (PAHC). Narrative interviews were conducted over a 4.5-year period and a phenomenological-hermeneutic method was used to interpret the text. The analysis resulted in one theme and four sub themes. Meanings of living the 'ups and downs' is symbolically understood as being captive in a roller-coaster ride, side by side. In togetherness one helps one another to integrate the illness into everyday life.When, the inevitable 'downs' come one alleviates them as much as possible and/or just endures them waiting to see if one outlives them. During the 'ups' one seizes every opportunity to live, doing the things one likes and discovering new possibilities in life. Sharing the safety belt on the 'roller coaster', offered by the PAHC team, evokes feelings of security.However, the safety belt is adjusted to the man with severe CHF leaving the wife without comfort and at times uncertain, especially in the deepest 'downs'. Our results indicate that the importance of medical treatment may increase over time. The ability to recognize symptoms and signs of deterioration may be facilitated by a trusted PAHC team's repeated questions about the condition. Furthermore it is important to offer support directly to the close relatives. PMID- 18386951 TI - 'I'm just not interested in eating': when nutrition becomes an issue in palliative care. AB - Coping with issues of hydration and nutrition are central when caring for a person at any stage of a life threatening or limiting illness. Particularly as death approaches, many practical and ethical issues may arise in consideration about how a person should be supported in assisting their nutrition and hydration. Support and education of the person themselves, their family and carers is vital in this process. This paper will use a case study and a take longitudinal view of illness, in addressing the nursing aspects of assisting a person's nutrition, firstly in the early stages of illness and then in considering nutritional needs in care towards the end of life. PMID- 18386952 TI - Nurses' experiences of clinical use of a quality of life instrument in palliative care. AB - This pilot study explored how nursing staff experienced clinical use of a quality of life (QoL) instrument as complement to their standard assessment interview. Twenty-six of 36 Registered Nurses working in one palliative care service responded in writing to six open questions.We found that nurses described positive features in using the QoL instrument related to how it supported their professional role, eased therapeutic contact with patients, and in terms of the outcomes of its use. Others noted difficulties incorporating the tool into the framework of their traditional nursing assessment, noting instrument and situation-specific limitations, with attitudes and comfort with the questionnaire affecting its use. Clinical use of QoL tools may be facilitated by emphasizing their role as a first screening assessment, and acknowledging and supporting the importance of clinical expertise and the patient-nurse relationship in further more focussed assessments and nursing care. PMID- 18386953 TI - Challenges and choices: an audit of the management of nausea, vomiting and bowel obstruction in metastatic ovarian cancer. AB - Women living with metastatic ovarian cancer experience many distressing symptoms, such as vomiting and bowel obstruction, which challenge the expertise of nurses working in Palliative Care to promote quality of life. These clinical challenges prompted us to audit our management of nausea/vomiting and bowel obstruction in a New Zealand Palliative Care service, with a focus on both pharmaceutical and non pharmaceutical interventions (e.g. dietary advice, relaxation strategies). The audit involved a retrospective chart review of 17 patients and identified the current clinical management of these symptoms in comparison with best practice guidelines. The results indicated many areas of symptom management were in line with current evidence-based practise; however the use of non-pharmaceutical interventions was limited. There were several implications for clinical practice, specifically supporting the use of electronic integrated patient management systems and a greater use of nonpharmaceutical (complementary) interventions. The audit process provided useful data for the authors to analyse current practice and promote more effective management of these distressing symptoms. PMID- 18386954 TI - Palliative care and vulnerability of self. PMID- 18386955 TI - Let's talk about sex: risky business for cancer and palliative care clinicians. AB - Few health professionals feel confident and comfortable when communicating with patients about the sexual and intimate changes that might occur after a diagnosis of cancer. Little research has focused on why health professionals find this type of patient communication so challenging. Drawing on data from a larger study examining issues of intimacy and sexuality from the perspectives of patients and health professionals in cancer and palliative care, this paper will present the health professional perspective. In the larger study a reflexive inquiry methodology enabled data to be collected through semi-structured participant interviews (n=82), a textual analysis of national and international clinical practice guidelines (n=33) and documented feedback from patients and health professionals attending educational forums where preliminary findings of the study were presented (n=15). In the part of the study reported here, a total of 32 health professionals recognised as members of a multidisciplinary team and working in cancer and or palliative care for a minimum of 12 months were interviewed. Results revealed that patient sexuality and intimacy was largely medicalised so that health professional discussions remained at the level of patient fertility, contraception, erectile or menopausal status. Many unchecked assumptions about patient sexuality were made by health professionals, based on the patient's age, diagnosis, culture, partnership and disease status. It was personally confronting and a 'risky' business to communicate about issues of patient intimacy and sexuality after cancer, particularly when the clinical setting emphasised medicalised, health professional driven and problem-based communication. Implications for practice will be discussed. PMID- 18386956 TI - Next of kin's feelings of guilt and shame in end-of-life care. AB - In this study the aim was to explore and describe next of kin's feelings of guilt and shame in end-of-life care via secondary analysis of 47 qualitative interviews. In the analysis categories and subcategories emerged which conveyed how the next of kin experienced guilt; not having done or talked enough, being absent at important events or making errors of judgement. Categories conveying feelings of shame were situations where the next of kin felt inferior, was ashamed on behalf of the dying person and when family conflicts became apparent. Receiving help and support in order to make the remaining period as pleasant as possible can facilitate the next of kin's sense of having fulfilled their duties and responsibilities and therein reduce feelings of guilt and shame. It is apparent that these feelings should be taken into account and the next of kin should receive support to increase their well-being during the remaining time in end-of-life care and the grieving period. PMID- 18386957 TI - Progress in understanding grief, complicated grief, and caring for the bereaved. AB - Grief occurs with loss of symbolically important connections and involves intense emotional reactions and changes to our experiences of self, the world, and the future. Individual responses reflect factors such as personality and life history, social context and cultural practices, and the symbolic magnitude of the loss. Grieving can be a relatively slow and uneven process, so applying prescriptive stages or goals to individuals' experiences can be unhelpful. Although most people are resilient in the face of loss and do not require special interventions, health professionals can contribute by empathic use of communication skills to facilitate the grieving process. A minority will struggle with their grief and experience prolonged, intense, or problematic reactions. Psychiatric comorbidities including depression and anxiety disorders can occur, and a distinct diagnosis of complicated grief disorder has been proposed. Health professionals can identify complicated grief reactions and ensure patients receive specialised treatment, including intensive grief therapy and medication where indicated. Assessment methods are summarised to assist health professionals in providing a continuum of care for those who are grieving. PMID- 18386958 TI - Bearing witness: an existential position in caring. AB - A basic assumption for the study is that perceiving a person's deepest needs and desires to be on hand for another person, and their attempt to do so, have, in an ontological sense, the power to bear witness of goodness and eternity. The study was based on a theoretical basis of a caring science view of suffering, as well as the ethics of the philosopher Levinas. The aim was to explore and clinically validate nuances of witnessing as a caring act.A Socratic dialogue was performed and an interpretive (hermeneutic) method was employed in this study. The Socratic dialogue with four nurses in palliative care focused on and analysed one clinical example of witnessing in palliative care. As basis for the findings are the participating nurses jointly formulated assumptions on the subject: To be a witness you have to be with the patient and refer back to him or her what you have seen; but also to act in accordance with what you have perceived. In the moment you witness, a window is opened onto the unknown; you become vulnerable as a caregiver and require courage. Being a witness encompasses existential and spiritual aspects; being a fellow human being, having a heart to heart relationship is a wilful act on the part of the nurse. Our theoretical discussion focuses on the language of the body, courage as a bridge to an existential encounter and the alleviation of patients' suffering through caregivers' witnessing. A conclusive aspect is that being a witness may bring a new understanding of life in the face of death and suffering. The existential position of being a witness requires the caregiver to be courageous because of its transformative prospect, but may utterly enrich both parties' inner life of shared meaning. PMID- 18386960 TI - Leading the way: innovative health promoting palliative care. PMID- 18386959 TI - Karmic quest: Thai family caregivers promoting a peaceful death for people with AIDS. AB - This paper reports the constructions of karma by four Thai family caregivers living with a dying person with AIDS in Southern Thailand. These four families form a subset of a larger ethnographic case study exploring the experiences of families living with a relative with AIDS. Serial interviews, observations, and field journal were used as data collection methods with the four families. The findings indicated that the karmic quest is a dominant theme in the narratives of these families caring for their loved ones dying with AIDS. The 'calm and peaceful' death that is described in the palliative care literature equated with their desire for the Buddhist philosophy of a harmonious death. The families used the law of karma and reincarnation as their main frame of reference and mobilised their religious resources to create meaning and purpose. Karmic healing activities were aimed at ending suffering, promoting a peaceful and calm death and ensuring a better life in the next one. The findings are important for the development of palliative nursing practice in Thailand by acknowledging religious and cultural values to promote peaceful death. PMID- 18386961 TI - The turning point: identifying end-of-life care in everyday health care practice. AB - There is a broad consensus among international policy statements that care provided at end-of-life should be different from care provided during other periods of life which assumes a turning point which reflects the onset of the dying process and the associated care reorientation. Few studies derived from empirical data have described this turning point from a clinical perspective. With the purpose to increase the clinical understanding about the nature of such turning points this study explores this phenomenon as determined from health-care records of a representative sample of adults who accessed the public health care system, and who died in the County of Vastra Gotaland, Sweden, during 2001 (n=229). Record entries reflecting a turning point, that is recognition of the patients as being at the end-of-life and a message about a switch from curative, life-extending or rehabilitative care to an emphasis on palliation, were found in slightly less than 70% of the records while slightly more than 30% lacked such descriptions. The record entries were analyzed with content analysis. As a whole, the descriptions were largely relying on a biomedical discourse and typically put in terms of signs of declining processes, undefined serious condition, prognostications of fatal outcome or justifications of given up active treatments. The procedures which followed the recognition of a turning point revealed that nurses and physicians typically emphasize procedures directed to the patients' families, on re-orientation of treatments and on approaches to further care. The picture of the patients' last time of life as reflected from these record entries is that end-of-life is described in biomedical and factual oriented terms. The end-of-life care was broadly described in vague terms with lack of concrete care plans. Holistic and good end-of-life care as revealed in this study is indistinguishable, highly embedded in a medical model of care and, hence, an abstract and unknown territory without specific content. PMID- 18386963 TI - From concept to care: enabling community care through a health promoting palliative care approach. AB - The aim of this paper was to implement and evaluate health promoting palliative care projects through partnerships formed within local communities in the Hume Region,Victoria, Australia. Health promoting palliative care is a public health approach to palliative care developed by Kellehear (1999). This article reports on a project in which the Hume Regional Palliative Care Service in Victoria, Australia sought to implement the approach through partnerships formed with a range of community groups and service agencies in their region. Funding was provided over a two-year period by the 'Caring Communities Program' of the Australian Commonwealth Department of Health and Ageing. It was hoped that the project overall would increase community capacity to participate in the care of members of the community living with dying and loss. The project developed implementation strategies and health promotion project guidelines, as outlined below. During the two-year period the palliative care service entered in a number of new partnerships with community groups and other health services. At first much of the initiative in forming these partnerships came from the palliative care service, but as the project progressed the initiative came increasingly from community partners. PMID- 18386962 TI - The influence of environment in palliative care: supporting or hindering experiences of 'at-homeness'. AB - Florence Nightingale stated that the art of nursing is to provide an environment in which the patient is in the best position for nature to act upon, and Martha Rogers, in turn, emphasized that one part in nursing is to pattern the environment into a place where healing conditions are optimal. This paper presents a preliminary conceptual framework that describes the influence of environment in palliative care. Based on previously published studies, a conceptual framework describing the influence of environments in palliative care was developed consisting of an atmosphere of hospitality; an atmosphere of safety; and, an atmosphere of 'everydayness'. The framework describes that the atmosphere is created in the meeting between the person's needs/expectations and the environment, an atmosphere that can influence experiential outcomes of 'at homeness' or homelessness in the palliative care setting. In conclusion, this preliminary conceptual framework may contribute to nursing practice by providing a conceptual basis for actively reflecting on and evaluating the atmosphere in palliative care settings. PMID- 18386964 TI - Advance care planning in residential aged care facilities. AB - A person's right to self-determination in health care is now a fundamental principle of health care provision. As a consequence Advance Care Planning (ACP) is a critical issue in health care including palliative care as it is seen as a way of ensuring a person's right to participate not only in future health care choices but in end of life decision making. To date there have been few reports of successful ACP programs. In 2004 and 2005 a program of advance care planning, known as Respecting Patient Choices was introduced in 17 residential aged care facilities in metropolitan Melbourne, Australia. This program consisted of a system-wide approach that included education for aged care staff; information for residents and families as well as changes to organisational processes to ensure that residents have opportunity for self-determination in their future health care. This paper will report on the lessons learnt from this implementation and identify strategies which foster sustainability of ACP. PMID- 18386966 TI - PROFALIGN algorithm identifies the regions containing folding determinants by scoring pairs of hydrophobic profiles of remotely related proteins. AB - Profile comparison methods have been shown to be very powerful in creating accurate alignments of protein sequences, especially in the case of remotely related proteins (RRP). These methods take advantage of the observation that hydrophobic profiles are more conserved than the corresponding amino acid sequences. Here, we present the PROFALIGN algorithm, which allows one to perform a detailed comparative analysis, at both local and global levels of two protein sequence profiles. The user can either choose among four different hydrophobic scales (Miyazawa-Jernigan, Eisenberg, Engelman-Steiz, and Kyte-Doolittle) or can add a personal scale. The interface is designed for a wide range of users, including those who are not involved in protein research. It allows one to vary the alignment parameters (such as gap penalties, embedding, and profile smoothness). Secondary structure propensity is added as an optional alignment filter. Similar segments of two proteins are singled out on the basis of score. We have tested the algorithm with different Src homology 3 (SH3) domain fragments sharing low sequence homology but very similar three-dimensional (3D) structures. By using the Miyazawa-Jernigan hydrophobic scale, PROFALIGN was able to detect the strong correlation between the regions that are known to be crucial for SH3 transition state topology. PROFALIGN seems able to identify most of the mutual alignment of structures on the basis of their hydrophobic profiles, delimiting the regions containing the key determinants of folding. Therefore, the present methodology may be useful for the detection of the most structurally relevant positions inside remote related proteins. PMID- 18386967 TI - Isolations of Jamestown Canyon virus (Bunyaviridae: Orthobunyavirus) from field collected mosquitoes (Diptera: Culicidae) in Connecticut, USA: a ten-year analysis, 1997-2006. AB - Jamestown Canyon virus (JCV) (Bunyaviridae: Orthobunyavirus) is a mosquito-borne zoonosis belonging to the California serogroup. It has a wide geographic distribution, occurring throughout much of temperate North America. White-tailed deer, Odocoileus virginianus are the principal amplification hosts, and boreal Aedes and Ochlerotatus mosquitoes are the primary vectors. A 10-year study was undertaken to identify potential mosquito vectors in Connecticut, quantify seasonal prevalence rates of infection, and define the geographic distribution of JCV in the state as a function of land use and white-tailed deer populations, which have increased substantially over this period. Jamestown Canyon virus was isolated from 22 mosquito species. Five of them, Ochlerotatus canadensis, Oc. cantator, Anopheles punctipennis, Coquillettidia perturbans, and Oc. abserratus were incriminated as the most likely vectors, based on yearly isolation frequencies and the spatial geographic distribution of infected mosquitoes. Jamestown Canyon virus was isolated from Oc. canadensis more consistently and from a greater range of collection sites than any other species. Frequent virus isolations were also made from Aedes cinereus, Aedes vexans, and Oc. sticticus, and new North American isolation records were established for Anopheles walkeri, Culex restuans, Culiseta morsitans, Oc. sticticus, Oc. taeniorhynchus, and Psorophora ferox. Other species from which JCV was isolated included C. melanura, Oc. aurifer, Oc. communis, Oc. excrucians, Oc. provocans, Oc. sollicitans, Oc. stimulans, Oc. triseriatus, and Oc. trivittatus. Jamestown Canyon virus was widely distributed throughout Connecticut and found to consistently circulate in a diverse array of mosquito vectors. Infected mosquitoes were collected from June through September, and peak infection rates paralleled mosquito abundance from mid-June through mid-July. Infection rates in mosquitoes were consistent from year to year, and overall virus activity was directly related to local mosquito abundance. Infected mosquitoes were equally distributed throughout the state, irrespective of land use, and infection rates were not directly associated with the abundance of white-tailed deer, possibly because of their saturation throughout the region. PMID- 18386969 TI - Profile: Interview with Rajeev Venkayya, MD, Director, Global Health Delivery, Bill & Melinda Gates Foundation, and Former Special Assistant to the President and Senior Director for Biodefense, White House Homeland Security Council. PMID- 18386970 TI - An evaluation of the role and effectiveness of institutional biosafety committees in providing oversight and security at biocontainment laboratories. AB - Institutional biosafety committees (IBCs) have been charged with the oversight and review of biosafety at thousands of biocontainment labs nationwide, hundreds of which are high-level BSL-3 and BSL-4 labs. In light of the recent rapid proliferation of BSL-3 and BSL-4 facilities and the increases in research in the areas of biodefense, select agents, recombinant DNA, and synthetic biology and dual-use research, questions have been raised about whether IBCs are fulfilling their oversight responsibilities. This article reviews information on the responsibilities and expectations of IBCs as currently constituted and provides an analysis of IBC performance from survey data of hundreds of research institutions over the past several years. The findings highlight serious ongoing problems with IBCs' adherence to NIH Guidelines. This raises questions about the current voluntary governance framework as an effective system to monitor and oversee U.S. research facilities, including high-containment facilities, and their research activities. The findings strongly suggest the need for immediate improvement or replacement of the IBC system. PMID- 18386971 TI - Can virtual reality be used to conduct mass prophylaxis clinic training? A pilot program. AB - OBJECTIVE: To create and evaluate a pilot bioterrorism defense training environment using virtual reality technology. METHODS: The present pilot project used Second Life, an internet-based virtual world system, to construct a virtual reality environment to mimic an actual setting that might be used as a Strategic National Stockpile (SNS) distribution site for northern California in the event of a bioterrorist attack. Scripted characters were integrated into the system as mock patients to analyze various clinic workflow scenarios. Users tested the virtual environment over two sessions. RESULTS: Thirteen users who toured the environment were asked to complete an evaluation survey. Respondents reported that the virtual reality system was relevant to their practice and had potential as a method of bioterrorism defense training. CONCLUSIONS: Computer simulations of bioterrorism defense training scenarios are feasible with existing personal computer technology. The use of internet-connected virtual environments holds promise for bioterrorism defense training. Recommendations are made for public health agencies regarding the implementation and benefits of using virtual reality for mass prophylaxis clinic training. PMID- 18386972 TI - Lidars: a key component of urban biodefense. AB - A biological aerosol attack in a city could infect tens of thousands of people. In the absence of announcements by the attacker or detection by present point detection systems, victims would be unaware of their exposure prior to developing symptoms. Since infections are most effectively countered before the onset of symptoms, detection technologies that provide early awareness of an attack should be given high priority. Current biological point detection (BPD) systems collect environmental air samples and then analyze them in laboratories so as to permit detection within 12-36 hours of an attack. Improvements in the pipeline may reduce this lag time to as little as a few hours. However, BPD systems have inherent weaknesses when used to detect and respond to an aerosol attack. The likelihood of a limited number of BPD systems intercepting an aerosol plume in a vast attack space may be low. Moreover, BPD systems do not provide critical information needed for response, such as the source location, precise time, and geographic reach of an attack. The missing information would invaluably guide prophylaxis distribution, identification of contaminated areas, and criminal apprehension. This article describes how lidars used for real-time observation of aerosol plumes could complement BPD systems by providing fine-scale spatial and temporal information. A lidar system also could be used to corroborate positive BPD system results, to improve reaction to positive results, and/or to provide an independent basis for low-regret protective steps. Lidar systems can resolve key biodefense challenges, and this article describes three compatible concepts of operations. Leveraging lessons from a lidar system now operating at the Pentagon, a test of an expanded lidar network would provide immediate protection for key Washington, DC, assets, demonstrate the synergy of BPD systems and lidars, and provide a test bed for research to improve lidar's shortcomings. PMID- 18386974 TI - Economic aspects of agricultural and food biosecurity. AB - Concerns about biosecurity in the food system raise a variety of issues about how the system is presently organized, why it might be vulnerable, what we could reasonably do to better secure it, and the costs of doing so. Emphasizing the role of incentives in efficient resource allocation, this article considers economic dimensions of three aspects of the general problem. One is the global problem, or the way biosecurity measures can affect how countries relate to each other and the global consequences that result. Another is how to best manage the immediate aftermath of a realized threat in order to minimize damage. The third is how to seek to prevent realization of the threat. Some policy alternatives are presented. PMID- 18386973 TI - Communicating information in an emergency preparedness pill distribution campaign. AB - OBJECTIVES: Public health agencies are often tasked with the development and execution of interventions, but the communication strategy and its impact on the effectiveness of an intervention is often not evaluated or incorporated by local and state health agencies. The primary objective of this study was to determine the impact of information from various sources on knowledge about an emergency preparedness public health intervention involving the mass distribution of medicine. METHODS: The study used validated written mail surveys containing 12 simple knowledge-based questions. One-way ANOVA, the Studentized Newman-Keuls (SNK) test, logistic regression, and multiple regression were used to evaluate the data. RESULTS: Reading an educational fact sheet or receiving job training were the highest predictive variables for correct responses on the survey among all groups. Commercial media were found to potentially diminish comprehension among survey respondents. There was significant variability in knowledge among different groups surveyed, ranging from an average of 15% to 74% correct responses on the survey. CONCLUSIONS: This study found that job training and fact sheets that are delivered directly to the intended recipients are very effective at enhancing knowledge among the general public and emergency responders. Conversely, we found that commercial media, such as television, may be detrimental to educating the public about important public health interventions. The internet was not widely used by the survey respondents to obtain information; this raises questions regarding the usefulness of websites for emergency preparedness education. PMID- 18386975 TI - Panalysis: a new spreadsheet-based tool for pandemic planning. AB - Publicly available influenza modeling tools are of limited use to hospitals and local communities in planning for a severe pandemic. We developed Panalysis, a new tool to estimate the likely healthcare consequences of a pandemic and to aid hospitals in the development of mitigation and response strategies. By way of example, we demonstrate how Panalysis can be used to plan for a 1918-like flu pandemic. We discuss potential future applications of this tool. PMID- 18386976 TI - What not to say: risk communication for botulism. AB - This formative research study used qualitative methods to test the suitability of messages about botulism for the general public. Nine focus group interviews and 27 cognitive interviews were conducted with diverse audiences to pretest radio, television, and fact sheet messages predicated on a hypothetical terrorist attack using botulinum toxin. Narrative data were collected, transcribed, coded, and analyzed using content domains based on risk and health communication theories. While participants accepted the need for materials, the messages produced contained images and references describing botulism as a toxin-caused illness spread both by food and water contamination as well as by airborne means. The audience's lack of understanding of the term toxin and an imperfect understanding of airborne transmission of a toxic substance meant that some people interpreted botulism as being an infectious disease rather than a type of poisoning. The communication materials did not clearly show how the set of botulism symptoms are unique and described the anti-toxin as "not a cure," thus compounding the audience's misunderstanding. Using models from cognitive and developmental psychology, our findings were interpreted to show that certain terms evoke or elicit long-held conceptual frameworks that lay audiences use to explain medical phenomena. Relevant to botulism, poisoning events are distinct from infectious diseases, but prepared messages did not reinforce these distinctions. Ignoring how people organize preexisting health information when trying to communicate new information is a prescription for failure, especially in a crisis risk communication scenario. Findings from this study have been used by the Centers for Disease Control and Prevention to reformulate pre-event crisis risk communication materials for botulism. PMID- 18386977 TI - Response to Epstein's commentary. PMID- 18386978 TI - Countermeasure R&D funding debate. PMID- 18386979 TI - Stochastic synchrony of chaos in a pulse-coupled neural network with both chemical and electrical synapses among inhibitory neurons. AB - The synchronous firing of neurons in a pulse-coupled neural network composed of excitatory and inhibitory neurons is analyzed. The neurons are connected by both chemical synapses and electrical synapses among the inhibitory neurons. When electrical synapses are introduced, periodically synchronized firing as well as chaotically synchronized firing is widely observed. Moreover, we find stochastic synchrony where the ensemble-averaged dynamics shows synchronization in the network but each neuron has a low firing rate and the firing of the neurons seems to be stochastic. Stochastic synchrony of chaos corresponding to a chaotic attractor is also found. PMID- 18386980 TI - Normalization enables robust validation of disparity estimates from neural populations. AB - Binocular fusion takes place over a limited region smaller than one degree of visual angle (Panum's fusional area), which is on the order of the range of preferred disparities measured in populations of disparity-tuned neurons in the visual cortex. However, the actual range of binocular disparities encountered in natural scenes extends over tens of degrees. This discrepancy suggests that there must be a mechanism for detecting whether the stimulus disparity is inside or outside the range of the preferred disparities in the population. Here, we compare the efficacy of several features derived from the population responses of phase-tuned disparity energy neurons in differentiating between in-range and out of-range disparities. Interestingly, some features that might be appealing at first glance, such as the average activation across the population and the difference between the peak and average responses, actually perform poorly. On the other hand, normalizing the difference between the peak and average responses results in a reliable indicator. Using a probabilistic model of the population responses, we improve classification accuracy by combining multiple features. A decision rule that combines the normalized peak to average difference and the peak location significantly improves performance over decision rules based on either measure in isolation. In addition, classifiers using normalized difference are also robust to mismatch between the image statistics assumed by the model and the actual image statistics. PMID- 18386983 TI - Understanding Parkinsonian handwriting through a computational model of basal ganglia. AB - Handwriting in Parkinson's disease (PD) is typically characterized by micrographia, jagged line contour, and unusual fluctuations in pen tip velocity. Although PD handwriting features have been used for diagnostics, they are not based on a signaling model of basal ganglia (BG). In this letter, we present a computational model of handwriting generation that highlights the role of BG. When PD conditions like reduced dopamine and altered dynamics of the subthalamic nucleus and globus pallidus externa subsystems are simulated, the handwriting produced by the model manifested characteristic PD handwriting distortions like micrographia and velocity fluctuations. Our approach to PD modeling is in tune with the perspective that PD is a dynamic disease. PMID- 18386982 TI - Dynamic moment analysis of the extracellular electric field of a biologically realistic spiking neuron. AB - Based on the membrane currents generated by an action potential in a biologically realistic model of a pyramidal, hippocampal cell within rat CA1, we perform a moment expansion of the extracellular field potential. We decompose the potential into both inverse and classical moments and show that this method is a rapid and efficient way to calculate the extracellular field both near and far from the cell body. The action potential gives rise to a large quadrupole moment that contributes to the extracellular field up to distances of almost 1 cm. This method will serve as a starting point in connecting the microscopic generation of electric fields at the level of neurons to macroscopic observables such as the local field potential. PMID- 18386984 TI - Algorithms for sparse nonnegative Tucker decompositions. AB - There is a increasing interest in analysis of large-scale multiway data. The concept of multiway data refers to arrays of data with more than two dimensions, that is, taking the form of tensors. To analyze such data, decomposition techniques are widely used. The two most common decompositions for tensors are the Tucker model and the more restricted PARAFAC model. Both models can be viewed as generalizations of the regular factor analysis to data of more than two modalities. Nonnegative matrix factorization (NMF), in conjunction with sparse coding, has recently been given much attention due to its part-based and easy interpretable representation. While NMF has been extended to the PARAFAC model, no such attempt has been done to extend NMF to the Tucker model. However, if the tensor data analyzed are nonnegative, it may well be relevant to consider purely additive (i.e., nonnegative) Tucker decompositions). To reduce ambiguities of this type of decomposition, we develop updates that can impose sparseness in any combination of modalities, hence, proposed algorithms for sparse nonnegative Tucker decompositions (SN-TUCKER). We demonstrate how the proposed algorithms are superior to existing algorithms for Tucker decompositions when the data and interactions can be considered nonnegative. We further illustrate how sparse coding can help identify what model (PARAFAC or Tucker) is more appropriate for the data as well as to select the number of components by turning off excess components. The algorithms for SN-TUCKER can be downloaded from Morup (2007). PMID- 18386985 TI - Random neural networks with synchronized interactions. AB - Large-scale distributed systems, such as natural neuronal and artificial systems, have many local interconnections, but they often also have the ability to propagate information very fast over relatively large distances. Mechanisms that enable such behavior include very long physical signaling paths and possibly saccades of synchronous behavior that may propagate across a network. This letter studies the modeling of such behaviors in neuronal networks and develops a related learning algorithm. This is done in the context of the random neural network (RNN), a probabilistic model with a well-developed mathematical theory, which was inspired by the apparently stochastic spiking behavior of certain natural neuronal systems. Thus, we develop an extension of the RNN to the case when synchronous interactions can occur, leading to synchronous firing by large ensembles of cells. We also present an O(N3) gradient descent learning algorithm for an N-cell recurrent network having both conventional excitatory-inhibitory interactions and synchronous interactions. Finally, the model and its learning algorithm are applied to a resource allocation problem that is NP-hard and requires fast approximate decisions. PMID- 18386986 TI - Encoding and decoding spikes for dynamic stimuli. AB - Naturally occurring sensory stimuli are dynamic. In this letter, we consider how spiking neural populations might transmit information about continuous dynamic stimulus variables. The combination of simple encoders and temporal stimulus correlations leads to a code in which information is not readily available to downstream neurons. Here, we explore a complex encoder that is paired with a simple decoder that allows representation and manipulation of the dynamic information in neural systems. The encoder we present takes the form of a biologically plausible recurrent spiking neural network where the output population recodes its inputs to produce spikes that are independently decodeable. We show that this network can be learned in a supervised manner by a simple local learning rule. PMID- 18386987 TI - Simultaneous approximations of polynomials and derivatives and their applications to neural networks. AB - We have constructed one-hidden-layer neural networks capable of approximating polynomials and their derivatives simultaneously. Generally, optimizing neural network parameters to be trained at later steps of the BP training is more difficult than optimizing those to be trained at the first step. Taking into account this fact, we suppressed the number of parameters of the former type. We measure degree of approximation in both the uniform norm on compact sets and the Lp-norm on the whole space with respect to probability measures. PMID- 18386988 TI - Optimizing one-shot learning with binary synapses. AB - A network of excitatory synapses trained with a conservative version of Hebbian learning is used as a model for recognizing the familiarity of thousands of once seen stimuli from those never seen before. Such networks were initially proposed for modeling memory retrieval (selective delay activity). We show that the same framework allows the incorporation of both familiarity recognition and memory retrieval, and estimate the network's capacity. In the case of binary neurons, we extend the analysis of Amit and Fusi (1994) to obtain capacity limits based on computations of signal-to-noise ratio of the field difference between selective and non-selective neurons of learned signals. We show that with fast learning (potentiation probability approximately 1), the most recently learned patterns can be retrieved in working memory (selective delay activity). A much higher number of once-seen learned patterns elicit a realistic familiarity signal in the presence of an external field. With potentiation probability much less than 1 (slow learning), memory retrieval disappears, whereas familiarity recognition capacity is maintained at a similarly high level. This analysis is corroborated in simulations. For analog neurons, where such analysis is more difficult, we simplify the capacity analysis by studying the excess number of potentiated synapses above the steady-state distribution. In this framework, we derive the optimal constraint between potentiation and depression probabilities that maximizes the capacity. PMID- 18386989 TI - Long-term visual object recognition memory in aged rats. AB - Aging is associated with memory impairments, but the neural bases of this process need to be clarified. To this end, behavioral protocols for memory testing may be applied to aged animals to compare memory performances with functional and structural characteristics of specific brain regions. Visual object recognition memory can be investigated in the rat using a behavioral task based on its spontaneous preference for exploring novel rather than familiar objects. We found that a behavioral task able to elicit long-term visual object recognition memory in adult Long-Evans rats failed in aged (25-27 months old) Wistar rats. Since no tasks effective in aged rats are reported in the literature, we changed the experimental conditions to improve consolidation processes to assess whether this form of memory can still be maintained for long term at this age: the learning trials were performed in a smaller box, identical to the home cage, and the inter trial delays were shortened. We observed a reduction in anxiety in this box (as indicated by the lower number of fecal boli produced during habituation), and we developed a learning protocol able to elicit a visual object recognition memory that was maintained after 24 h in these aged rats. When we applied the same protocol to adult rats, we obtained similar results. This experimental approach can be useful to study functional and structural changes associated with age related memory impairments, and may help to identify new behavioral strategies and molecular targets that can be addressed to ameliorate memory performances during aging. PMID- 18386990 TI - Homeostatic cytokines and expansion of regulatory T cells accompany thymic impairment in children with Down syndrome. AB - Down syndrome (DS), the most common chromosomal abnormality in humans, is characterized by precocious immunologic aging that results, among other things, in alterations of B and T lymphocyte subsets and natural killer cells, defective phagocytosis, and chemotaxis of polymorphonuclear leukocytes. We studied 30 children affected by DS, compared them to 29 healthy controls, and evaluated the functionality of the thymus (by measuring the amount of lymphocytes that express the signal-joint T cell receptor rearrangement excision circles [sj-TREC+]), the plasma levels of interleukin (IL)-7 and IL-15, the proliferative T cell response to these cytokines, the expression of the alpha chain of the IL-7 receptor (CD127), the extrathymic differentiation of T lymphocytes, and the presence of natural regulatory T cells (Tregs) in peripheral blood. We found that DS children had a significantly lower number of sj-TREC+ lymphocytes, the levels of which were strongly correlated with age. We found higher plasma levels of IL-7 and IL 15 than in healthy controls, and a higher proliferative T cell response to IL-15. DS children also showed a lower percentage of CD4(+) cells and profound alterations of T cell differentiation, along with increased amount of Tregs and of cells expressing markers of apoptosis. We can thus hypothesize that the precocious thymic involution occurring in DS is mirrored by a high production of IL-7 and IL-15, which is crucial for cell survival and proliferation. The complex alterations present in the periphery are likely the result of a compensatory mechanism: the overproduction of homeostatic cytokines could be a reaction to the impaired intrathymic production of T lymphocytes and/or to the expansion of Treg in the periphery, and could be required to allow the survival of T cells. PMID- 18386991 TI - Karyotypically normal and abnormal human embryonic stem cell lines derived from PGD-analyzed embryos. AB - Although a normal karyotype is generally a requirement for stem cell lines, new applications are likely to emerge for stem cells with defined chromosomal aneuploidies. We therefore investigated the use of embryos found to be aneuploid on biopsy followed by preimplantation genetic diagnosis (PGD) with fluorescent in situ hybridization (FISH), and developmentally arrested embryos for stem cell derivation. Eleven stem cell lines were obtained from 41 embryos in 36 cultures, with higher success rate achieved from PGD-analyzed, developmentally advanced embryos (45%) than from clinically unsuitable non-PGD embryos (13%). The resulting stem cell lines were karyotyped, and surprisingly, six of the nine lines from aneuploid embryos as well as both lines from non-PGD embryos were karyotypically normal. Three lines from PGD embryos were aneuploid exhibiting trisomy 5, trisomy 16, and an isochromosome 13, respectively. None of the aneuploid lines presented the same anomally as the original PGD analysis. Our study has three important implications. First, we confirm the ability to produce stem cell lines from PGD-tested embryos as well as developmentally abnormal embryos, offering specialty stem cell lines for research into the clinically important aneuploidies. Second, we observe that stem cell derivation from apparently aneuploid embryos is often thwarted by underlying mosaicism and emerging dominance of the stem cell line by karyotypically normal cells. The corollary, however, is that regular production of normal stem cell lines from developmentally abnormal embryos ordinarity discarded opens a new source of embryos for stem cells, whether for research or for eventual therapeutic use within the donating families. PMID- 18386992 TI - Sequential genetic modification of the hprt locus in human ESCs combining gene targeting and recombinase-mediated cassette exchange. AB - Genetic modification of human embryonic stem cells (hESCs) will be an essential tool to allow full exploitation of these cells in regenerative medicine and in the study of hESC biology. Here we report multiple sequential modifications of an endogenous gene (hprt) in hESCs. A selectable marker flanked by heterospecific lox sites was first introduced by homologous recombination (HR) into the hprt gene. In a subsequent step, exchange of the selectable marker with another cassette was achieved by recombinase-mediated cassette exchange (RMCE). We show that 100% of the recovered clones were the result of RMCE using a promoter trap strategy at the hprt locus. hprt-targeted H1 cells maintained a diploid karyotype and expressed hESC surface markers before and after RMCE. Finally, we report a double replacement strategy using two sequential gene targeting steps resulting in the targeted correction of an hprt-mutated hESC line. PMID- 18386994 TI - Tracking moving optima using Kalman-based predictions. AB - The dynamic optimization problem concerns finding an optimum in a changing environment. In the field of evolutionary algorithms, this implies dealing with a time-changing fitness landscape. In this paper we compare different techniques for integrating motion information into an evolutionary algorithm, in the case it has to follow a time-changing optimum, under the assumption that the changes follow a nonrandom law. Such a law can be estimated in order to improve the optimum tracking capabilities of the algorithm. In particular, we will focus on first order dynamical laws to track moving objects. A vision-based tracking robotic application is used as testbed for experimental comparison. PMID- 18386995 TI - Automated discovery of local search heuristics for satisfiability testing. AB - The development of successful metaheuristic algorithms such as local search for a difficult problem such as satisfiability testing (SAT) is a challenging task. We investigate an evolutionary approach to automating the discovery of new local search heuristics for SAT. We show that several well-known SAT local search algorithms such as Walksat and Novelty are composite heuristics that are derived from novel combinations of a set of building blocks. Based on this observation, we developed CLASS, a genetic programming system that uses a simple composition operator to automatically discover SAT local search heuristics. New heuristics discovered by CLASS are shown to be competitive with the best Walksat variants, including Novelty+. Evolutionary algorithms have previously been applied to directly evolve a solution for a particular SAT instance. We show that the heuristics discovered by CLASS are also competitive with these previous, direct evolutionary approaches for SAT. We also analyze the local search behavior of the learned heuristics using the depth, mobility, and coverage metrics proposed by Schuurmans and Southey. PMID- 18386996 TI - Finding alternatives and reduced formulations for process-based models. AB - This paper addresses the problem of model complexity commonly arising in constructing and using process-based models with intricate interactions. Apart from complex process details the dynamic behavior of such systems is often limited to a discrete number of typical states. Thus, models reproducing the system's processes in all details are often too complex and over-parameterized. In order to reduce simulation times and to get a better impression of the important mechanisms, simplified formulations are desirable. In this work a data adaptive model reduction scheme that automatically builds simple models from complex ones is proposed. The method can be applied to the transformation and reduction of systems of ordinary differential equations. It consists of a multistep approach using a low dimensional projection of the model data followed by a Genetic Programming/Genetic Algorithm hybrid to evolve new model systems. As the resulting models again consist of differential equations, their process-based interpretation in terms of new state variables becomes possible. Transformations of two simple models with oscillatory dynamics, simulating a mathematical pendulum and predator-prey interactions respectively, serve as introductory examples of the method's application. The resulting equations of force indicate the predator-prey system's equivalence to a nonlinear oscillator. In contrast to the simple pendulum it contains driving and damping forces that produce a stable limit cycle. PMID- 18386997 TI - Genetic algorithms for data-driven web question answering. AB - We present an evolutionary approach for the computation of exact answers to natural languages (NL) questions. Answers are extracted directly from the N-best snippets, which have been identified by a standard Web search engine using NL questions. The core idea of our evolutionary approach to Web question answering is to search for those substrings in the snippets whose contexts are most similar to contexts of already known answers. This context model together with the words mentioned in the NL question are used to evaluate the fitness of answer candidates, which are actually randomly selected substrings from randomly selected sentences of the snippets. New answer candidates are then created by applying specialized operators for crossover and mutation, which either stretch and shrink the substring of an answer candidate or transpose the span to new sentences. Since we have no predefined notion of patterns, our context alignment methods are very dynamic and strictly data-driven. We assessed our system with seven different datasets of question/answer pairs. The results show that this approach is promising, especially when it deals with specific questions. PMID- 18386998 TI - On complexity of optimal recombination for binary representations of solutions. AB - We consider the optimization problem of finding the best possible offspring as a result of a recombination operator in an evolutionary algorithm, given two parent solutions. The optimal recombination is studied in the case where a vector of binary variables is used as a solution encoding. By means of efficient reductions of the optimal recombination problems (ORPs) we show the polynomial solvability of the ORPs for the maximum weight set packing problem, the minimum weight set partition problem, and for linear Boolean programming problems with at most two variables per inequality, and some other problems. We also identify several NP hard cases of optimal recombination: the Boolean linear programming problems with three variables per inequality, the knapsack, the set covering, the p-median, and some other problems. PMID- 18387000 TI - Important role of the LKB1-AMPK pathway in suppressing tumorigenesis in PTEN deficient mice. AB - The LKB1 tumour suppressor phosphorylates and activates AMPK (AMP-activated protein kinase) when cellular energy levels are low, thereby suppressing growth through multiple pathways, including inhibiting the mTORC1 (mammalian target of rapamycin complex 1) kinase that is activated in the majority of human cancers. Blood glucose-lowering Type 2 diabetes drugs also induce LKB1 to activate AMPK, indicating that these compounds could be used to suppress growth of tumour cells. In the present study, we investigated the importance of the LKB1-AMPK pathway in regulating tumorigenesis in mice resulting from deficiency of the PTEN (phosphatase and tensin homologue deleted on chromosome 10) tumour suppressor, which drives cell growth through overactivation of the Akt and mTOR (mammalian target of rapamycin) kinases. We demonstrate that inhibition of AMPK resulting from a hypomorphic mutation that decreases LKB1 expression does not lead to tumorigenesis on its own, but markedly accelerates tumour development in PTEN(+/ ) mice. In contrast, activating the AMPK pathway by administration of metformin, phenformin or A-769662 to PTEN(+/-) mice significantly delayed tumour onset. We demonstrate that LKB1 is required for activators of AMPK to inhibit mTORC1 signalling as well as cell growth in PTEN-deficient cells. Our findings highlight, using an animal model relevant to understanding human cancer, the vital role that the LKB1-AMPK pathway plays in suppressing tumorigenesis resulting from loss of the PTEN tumour suppressor. They also suggest that pharmacological inhibition of LKB1 and/or AMPK would be undesirable, at least for the treatment of cancers in which the mTORC1 pathway is activated. Most importantly, our results demonstrate the potential of AMPK activators, such as clinically approved metformin, as anticancer agents, which will suppress tumour development by triggering a physiological signalling pathway that potently inhibits cell growth. PMID- 18387001 TI - Translational regulation of lipoprotein lipase in adipocytes: depletion of cellular protein kinase Calpha activates binding of the C subunit of protein kinase A to the 3'-untranslated region of the lipoprotein lipase mRNA. AB - Adipose LPL (lipoprotein lipase) plays an important role in regulating plasma triacylglycerols and lipid metabolism. We have previously demonstrated that PKCalpha (protein kinase Calpha) depletion inhibits LPL translation in 3T3-F442A adipocytes. Using in vitro translation experiments, the minimum essential region on the 3'UTR (3'-untranslated region) of LPL mRNA required for the inhibition of translation was identified as the proximal 39 nt. These results were confirmed by RNase protection analysis using cytoplasmic proteins isolated from the adipocytes treated with PKCalpha antisense oligomers and the LPL 3'UTR transcript (LPL 3'UTR nt: 1512-1640). The protein components involved in this RNA-binding interaction from PKCalpha depletion were passed through an affinity column containing a sequence of the LPL 3'UTR and, after Western blotting, the RNA-binding proteins were identified as the catalytic and the regulatory subunits of PKA (protein kinase A), Calpha and RIIbeta, and AKAP (A-kinase-anchoring protein) 121. This RNA inhibitory complex consisted of the same RNA-binding proteins that have been identified previously as mediators of LPL translational inhibition by PKA activation, suggesting that PKCalpha depletion inhibits LPL translation through PKA activation. In additional experiments, PKC depletion by prolonged PMA treatment or PKCalpha antisense oligomers resulted in an increase in PKA activity in 3T3-F442A adipocytes, comparable with PKA activation with adrenaline (epinephrine) treatment. These results demonstrate that LPL translational inhibition occurs through an RNA-binding complex involving PKA subunits and AKAP121, and this complex can be activated either through traditional PKA activation methods or through the depletion of PKCalpha. PMID- 18387002 TI - TGFbeta1 regulates endothelial cell spreading and hypertrophy through a Rac-p38 mediated pathway. AB - BACKGROUND INFORMATION: TGFbeta (transforming growth factor beta) is a multifunctional cytokine and a potent regulator of cell growth, migration and differentiation in many cell types. In the vascular system, TGFbeta plays crucial roles in vascular remodelling, but the signalling pathways involved remain poorly characterized. RESULTS: Using the model of porcine aortic endothelial cells, we demonstrated that TGFbeta stimulates cellular spreading when cells are on collagen I. TGFbeta-stimulated Rac1-GTP accumulation, which was associated with increased MAPK (mitogen-activated protein kinase) p38 phosphorylation. Furthermore, ectopic expression of a dominant-negative Rac mutant, or treatment of the cells with the p38 pharmacological inhibitor SB203580, abrogated TGFbeta induced cell spreading. Our results demonstrate for the first time that prolonged exposure to TGFbeta stimulates endothelial cell hypertrophy and flattening. Collectively, these data indicate that TGFbeta-induced cell spreading and increase in cell surface areas occurs via a Rac-p38-dependent pathway. CONCLUSIONS: The Rac-p38 pathway may have conceptual implications in pathophysiological endothelial cell responses to TGFbeta, such as wound healing or development of atherosclerotic lesions. PMID- 18387003 TI - Abstracts of the Perinatal Society of Australia and New Zealand 12th Annual Congress. April 20-23, 2008, Gold Coast, Australia. PMID- 18387004 TI - Risk prediction: a review of the literature. AB - This review of risk assessment and prediction literature briefly describes the historical and philosophical influences on the construct of dangerousness; chronicles the advances in research associated with the reconceptualization of dangerousness as risk; and describes current practice and research related to risk assessment, with particular attention to commonly used risk assessment tools and to existing issues and controversies. This cannot be considered a systematic review of the literature but a reflection of some of the key issues found in the literature. PMID- 18387005 TI - Violence education in nursing: critical reflection on victims' stories. AB - Violence against women is a major public health concern. This paper describes an educational strategy to increase nursing students' understanding of the experience of violence and to foster recognition and intervention with victims of violence. Students in an elective course were asked to critically reflect on the personal stories of victims/survivors of violence. The assignment provided four learning opportunities that include examination of societal myths on sexual victimization, understanding the lived experience of the victim, exploration of personal beliefs and values, and the relationship of the individual's experience to theoretical content of the course. Students gave permission for the use of quotes from papers to illustrate the learning opportunities. PMID- 18387006 TI - A participatory evaluation project to measure SANE nursing practice and adult sexual assault patients' psychological well-being. AB - This paper describes a collaborative project between a team of researcher evaluators and a Sexual Assault Nurse Examiner (SANE) program to develop an evaluation survey of SANE nursing practice and patient psychological well-being. Using a participatory evaluation model, we followed a six-step process to plan and conduct an evaluation of adult sexual assault patients treated in one Midwestern SANE program. Our collaborative team developed a logic model of "empowering care," which we defined as providing healthcare, support, and resources; treating survivors with dignity and respect; believing their stories; helping them reinstate control and choice; and respecting patients' decisions. We created a corresponding survey that can be administered to patients following exam procedures and tested it with N= 52 sexual assault victims. Results indicated that nursing practice was consistent with this empowering care philosophy as the overwhelming majority of patients reported positive psychological well-being outcomes. Implications for evaluating forensic nursing practice are discussed. PMID- 18387007 TI - Psychopathy: a confusing clinical construct. AB - Although psychopathy has traditionally been cited as a disorder of personality, confusion arises as the term is used interchangeably with the terms antisocial personality disorder and dissocial personality disorder, both of which are largely behaviorally based. This paper aims to provide an overview of the literature on the topic of psychopathy, which examines this conundrum. Included in the discussion are definitions of psychopathy, incidence, approaches to diagnosis, and the debates that surround causes, manifestations, and treatability. Experimental studies and theoretical papers have been included if considered to be informative and of relevance to forensic nursing practice. The review demonstrates that studies are fragmented and no clear consensus seems to emerge concerning any of the discussion areas or even the construct of psychopathy itself. It is concluded that further research is required in psychopathy as encountered in both institutional and community settings. Until complete clarification is provided by the research community, forensic nurses need to maintain positive views about their own role when working with people with this challenging condition and strive to maintain a therapeutic ward atmosphere. PMID- 18387008 TI - Retrieval of DNA from the faces of children aged 0-5 years: a technical note. AB - Approximately 21% of children suffer from some form of physical abuse. It is hypothesized that when an individual hits a child some of that person's DNA will be deposited onto the child's skin. As yet, no one has reported a method of sampling DNA from the skin of this vulnerable group of individuals. We have sampled DNA from several facial areas of 30 children aged 5 years of age and under. The results show that it is possible to swab the faces of this age group without distressing them or contaminating the samples. Additionally the results indicate that the DNA obtained is almost entirely that of the subject, with little nondonor DNA being observed. PMID- 18387009 TI - Forensic nursing science in 2008. PMID- 18387010 TI - Safeguarding attitudes. PMID- 18387011 TI - Toto's guide to reading and using research manuscripts for forensic nursing practice. PMID- 18387012 TI - One voice. PMID- 18387013 TI - Predisposing factors to phlebitis in patients with peripheral intravenous catheters: a descriptive study. AB - PURPOSE: The purpose of this study was to investigate the predisposing factors in the development of phlebitis in peripheral intravenous (IV) catheterization sites in patients treated with a variety of IV infusion solutions and drugs. DATA SOURCES: Systematic observation of 568 IV sites inserted for fluid infusion and drug administration in 355 patients in the Department of General Surgery of a University Hospital in Turkey. A data collection tool was based on standards established by the Infusion Nurses Society. Patients' infusion sites were monitored every 24 h during treatment and for 48 h after discontinuation of the IV. CONCLUSIONS: In contrast to the usual findings in the literature, the authors found that infusion through an infusion pump and insertion of catheters in the veins around the elbow increased the risk of phlebitis. Also, the number of times infusions were started led to an increased rate of phlebitis. However, conflicting results were obtained about the relation between phlebitis, gender, and catheter size. IMPLICATIONS FOR PRACTICE: Phlebitis causes sepsis, pain, additional diagnostic investigations, and treatments, and may lead to increased duration of hospitalization, patient's stress level, and financial burden, as well as increasing staff workload. Advanced practice nurses need to be aware of the factors that increase the likelihood of phlebitis and take appropriate measures to prevent it. PMID- 18387014 TI - Clinical practice algorithms: medication management to reduce fall risk in the elderly-part 4, anticoagulants, anticonvulsants, anticholinergics/bladder relaxants, and antipsychotics. PMID- 18387015 TI - Guidelines update on the prevention of heart disease in women. PMID- 18387016 TI - Family-centered collaborative negotiation: a model for facilitating behavior change in primary care. AB - PURPOSE: To describe a parent-child-based model that melds a family-centered interaction approach, Touchpoints, with brief negotiation strategies (an adaptation of motivational interviewing) to address health risks in children. An application of the model for addressing childhood overweight in the primary care setting is presented. DATA SOURCES: Selected research, theoretical, and clinical articles; national recommendations and guidelines; and a clinical case. CONCLUSIONS: Lifestyle health behaviors are learned and reinforced within the family; thus, changes to promote child health require family involvement. Interventions that engage parents and support parent-child relationships, while enhancing motivation and the abilities to change behavior, are recommended. IMPLICATIONS FOR PRACTICE: Primary care is an appropriate setting for addressing lifestyle health behaviors. A collaborative partnership, rather than a prescriptive manner, is advocated for primary care providers when working to facilitate health-promoting behavior. PMID- 18387017 TI - Empowering your patients in the fight against methicillin-resistant Staphylococcus aureus. AB - PURPOSE: To provide patient teaching points for primary care management and control of methicillin-resistant Staphylococcus aureus (MRSA) through application of the latest research regarding transmission of this bacteria. DATA SOURCES: Case reports, scientific literature, and the recommendations of expert professional groups. CONCLUSIONS: MRSA is a well studied yet continually evolving superbug. There is a paucity of literature regarding detailed home management and containment of MRSA. This review acknowledges the critical importance of patient education regarding MRSA infections and empowers patients with knowledge that can positively impact treatment outcomes. IMPLICATIONS FOR PRACTICE: Awareness of transmission modes and recognition of sources for relapse of infectious states can curb the spread of MRSA in the community. PMID- 18387018 TI - Mindfulness-based stress reduction: a literature review and clinician's guide. AB - PURPOSE: To provide nurse practitioners (NPs) with clinical research about Mindfulness-Based Stress Reduction (MBSR) and demonstrate its usefulness for reducing stress in a variety of populations. DATA SOURCES: A literature review was conducted using the following databases: EBSCO, Cinahl, Pschyline, and Medline. English language articles published between 2000 and 2006 in peer reviewed journals were reviewed. Search terms "mindfulness,""meditation," and "stress" were used. Additional information was obtained through select, reputable Internet sites. CONCLUSIONS: MBSR is an effective treatment for reducing stress and anxiety that accompanies daily life and chronic illness. MBSR is also therapeutic for healthcare providers, enhancing their interactions with patients. No negative side effects from MBSR have been documented. IMPLICATIONS FOR PRACTICE: MBSR is a safe, effective, integrative approach for reducing stress. Patients and healthcare providers experiencing stress or stress-related symptoms benefit from MBSR programs. NPs can safely and effectively use this intervention in a variety of patient populations. PMID- 18387019 TI - Sodium dietary restriction, knowledge, beliefs, and decision-making behavior of older females. AB - PURPOSE: The purposes of this qualitative/descriptive study were to (a) explore experiences and decision-making behaviors associated with adoption of a sodium restricted diet (SRD) among older women with hypertension or heart failure and (b) identify healthcare system and contextual factors that facilitate or impede adherence to SRD. DATA SOURCES: Participants were 33 single older women, aged 65 98 years, residing in three congregate living facilities in the high-risk "coronary valley" area of the United States. A semistructured interview format was employed with three focus groups. The audio-taped transcribed data were content analyzed for themes by the researchers with the assistance of ATLAS.Ti computer software. CONCLUSIONS: Predominant themes were lack of SRD education by healthcare providers, a desire for more information about sodium, including the use of alternative herbal seasonings, and large-print informational materials. Eating alone with no motivation to cook and share meals was a contextual barrier to healthy nutrition. IMPLICATIONS FOR PRACTICE: To prevent costly hospitalizations and rehospitalization from nonadherence to SRD, clinicians need to provide more structured SRD education supplemented with printed brochures. Exploring the client's nutritional social setting may improve SRD adherence. PMID- 18387020 TI - Obtaining a thorough sleep history and routinely screening for obstructive sleep apnea. AB - PURPOSE: To present a clinical case study of obstructive sleep apnea (OSA) and discuss a potential correlation between OSA, unexplained distal pain symptoms, and pyschoemotional concerns. DATA SOURCES: A review of the scientific literature was performed on OSA using the Cumulative Index of Nursing and Allied Health Literature and MEDLINE. CONCLUSIONS: OSA is potentially life threatening and can have serious consequences to a patient's health. Many of the obvious signs of OSA occur at night, and the symptoms of OSA may correspond to a variety of other diseases. Clinicians should recognize a possible correlation between OSA and unexplained distal pain symptoms as well as psychoemotional concerns. These clinically associated conditions may be less apparent but may dramatically affect quality of life. By improving recognition and treatment of OSA, morbidity and mortality can be reduced and quality of life can be improved for patients and their families. IMPLICATIONS FOR PRACTICE: It is imperative that clinicians are attentive and take detailed histories to recognize the clinical signs and symptoms of OSA, paying more attention to the less obvious symptomatology that may be significantly impacting quality of life. PMID- 18387022 TI - Childhood obesity in a South Florida elementary school. PMID- 18387023 TI - Changing nutrition standards in schools: the emerging impact on school revenue. AB - BACKGROUND: Although great focus has been placed on nutritional and other consequences of changes in food-related policies within schools, few reports exist describing the impact of such changes on school revenue. This review provides an overview of the few revenue-related studies published recently, as well as information from a sampling of state reports on the subject. METHODS: A systematic review of the literature was conducted. Four peer-reviewed papers and 3 state-based reports were identified that assessed the impact on revenues of either targeted policy changes or overarching, district-wide changes in food related policies. RESULTS: Thus far, few data exist to substantiate the concern that changes in nutrition standards in schools lead to a loss in total revenue. An interesting phenomenon of increased participation in the National School Lunch Program was noted in a number of reports and might play a role in buffering financial losses. CONCLUSIONS: A renewed focus on school policies related to health provides the opportunity for researchers to investigate how nutrition related policy change can affect, if at all, food service and overall school revenues. PMID- 18387024 TI - Relationship between physical disabilities or long-term health problems and health risk behaviors or conditions among US high school students. AB - BACKGROUND: This study explores the relationship between self-reported physical disabilities or long-term health problems and health risk behaviors or adverse health conditions (self-reported engagement in violent behaviors, attempted suicide, cigarette smoking, alcohol and other drug use, sexual activity, physical activity, dietary behaviors, self-reported overweight [based on height and weight], physical health, and mental health) among US high school students. METHODS: Data were from the Centers for Disease Control and Prevention's 2005 national Youth Risk Behavior Survey, a cross-sectional paper-and-pencil survey collected from a representative sample of public and private high school students (grades 9 through 12) in the United States. RESULTS: Significantly more students with physical disabilities or long-term health problems than without described their health as fair or poor and reported being in a physical fight, being forced to have sexual intercourse, feeling sad or hopeless, seriously considering and attempting suicide, cigarette smoking, using alcohol and marijuana, engaging in sexual activity, using computers 3 or more hours per day, and being overweight (for all, p < or = .05). For none of the health risk behaviors analyzed were the rates significantly lower among students with physical disabilities or long-term health problems than among other students. CONCLUSIONS: Young people who live with physical disabilities or long-term health problems may be at greater risk for poor health outcomes. Public health and school health programs, with guidance from health care providers, need to work with these adolescents and their families to develop and implement appropriate interventions, with particular emphasis on promoting mental health. PMID- 18387025 TI - The varied circumstances prompting requests for emergency contraception at school based clinics. AB - BACKGROUND: Little is known about the circumstances that prompt teenagers to request emergency contraception (EC). This evaluation was designed to refine the EC clinical protocol and improve pregnancy prevention efforts in high school based clinics by analyzing information on EC use and subsequent contraception use of EC patients. METHODS: Sites included 5 clinics located at mainstream, racially diverse, and urban high schools. Nursing staff documented all EC-related visits during the 2002-2003 school year on a standardized form. These forms and additional information were collected from chart reviews. RESULTS: EC was requested in 113 instances involving 91 students and dispensed in all but 4 instances. The most frequently reported circumstance prompting an EC request was the use of no protection (37.2%), followed by a condom mishap (27.4%) and questionable protection from a hormonal method (23.9%). Anxiety despite adequate protection from a hormonal method (9.7%) or a condom (1.8%) accounted for the remaining cases. Although the clinical protocol included a follow-up appointment 2 weeks after receipt of EC, 39.5% of EC users failed to keep this appointment. The chart review revealed that hormonal contraception was eventually initiated following two thirds (68.0%) of the EC instances that involved either no protection or only a condom. CONCLUSIONS: Because young EC seekers vary considerably in terms of sexual experience, contraceptive use consistency, and pregnancy risk, individualized risk assessment and counseling are essential. The EC visit may be an optimal time to encourage initiation or reinstitution of hormonal methods of birth control. PMID- 18387027 TI - The lack of motor vehicle occupant restraint use in children arriving at school. AB - BACKGROUND: Motor vehicle crashes are the leading cause of mortality for children aged 4-14 in the United States. Many children are driven daily to school, increasing their exposure to potential injury, especially if they are not appropriately restrained. Observing the level of motor vehicle occupant restraint (MVOR) use upon school arrival could lead to valuable information for the development of an injury prevention program targeted to this population. This study compares the rate of MVOR use upon arrival to elementary schools with that at regional intersections. METHODS: One hundred thirty-five census tracts in the Denver, Colorado, area were identified and combined into 5 regions. Within each region, 1 controlled intersection and 1 elementary school were observed. Observations were conducted for 1 hour at each of these locations. RESULTS: At intersections, 618 children in private vehicles were observed. MVOR use rate was 71.2% (440/618). At schools, 665 children in private vehicles were observed. MVOR use was 24.8% (165/665). Children arriving at school were less likely to be restrained than those observed at intersections (relative risk: 0.35, 95% CI: 0.30-0.40). CONCLUSIONS: Children at elementary schools showed substantially lower MVOR use rates than those observed at controlled intersections. Further investigation is imperative to determine the reason behind the low use of MVOR in children being transported to school. Based on the reasons elicited, interventions emphasizing the importance of always using MVOR can be developed and tailored to meet the educational needs of parents transporting their children to school in private vehicles. PMID- 18387026 TI - Factors influencing teachers' implementation of an innovative tobacco prevention curriculum for multiethnic youth: Project SPLASH. AB - BACKGROUND: The effectiveness of school-based tobacco use prevention programs depends on proper implementation. This study examined factors associated with teachers' implementation of a smoking prevention curriculum in a cluster randomized trial called Project SPLASH (Smoking Prevention Launch Among Students in Hawaii). METHODS: A process evaluation was conducted and a cross-condition comparison used to examine whether teacher characteristics, teacher training, external facilitators and barriers, teacher attitudes, and curriculum attributes were associated with the dose of teacher implementation in the intervention and control arms of the study. Data were collected from a total of 62 middle school teachers in 20 public schools in Hawaii, during the 2000-2001 and 2001-2002 school years. Sources included teacher questionnaires and interviews. Chi-square test and t test revealed that implementation dose was related to teachers' disciplinary backgrounds and skills and student enjoyment of the curriculum. RESULTS: Content analysis, within case, and cross-case analyses of qualitative data revealed that implementing the curriculum in a year long class schedule and high teacher self-efficacy supported implementation, while high perceived curriculum complexity was associated with less complete implementation. CONCLUSIONS: The results have implications for research, school health promotion practice, and the implementation of evidence-based youth tobacco use prevention curricula. PMID- 18387028 TI - Perceived competence in addressing student substance abuse: a national survey of middle school counselors. AB - BACKGROUND: Student substance abuse is a serious concern for middle school personnel. School counselors are most likely to deliver mental health services, including substance abuse, in school settings. However, limited research is available on the perceived competence of middle school counselors for addressing student substance abuse concerns. The main purpose of this study was to determine how middle school counselors perceive their training in 9 competence areas related to student substance abuse. A secondary aim of this study was to identify which training areas counselors indicate as being most needed to address student substance abuse. METHODS: A survey study was conducted that included a national sample of 283 middle school counselors. Analysis of variance was used to determine differences between 9 competency areas rated by counselors. Further analysis was conducted to determine which training areas were most important to counselors. RESULTS: Counselors varied in their perceived competence depending on the specific area of student substance abuse; however, they were clearly able to identify the most important areas of training needed. CONCLUSIONS: Overall, the findings from this study indicate that middle school counselors require more training in the area of student substance abuse. Specifically, this study provides information on the most important areas of training as identified by counselors. PMID- 18387029 TI - Prevalence of students with symptoms of depression among high school students in a district of western Turkey: an epidemiological study. AB - BACKGROUND: To determine the factors affecting the prevalence of depression and also to present some pertinent comments concerning prevention of depression among high school students. This study was deemed important and relevant due to the increasing importance of depression among high school students. METHODS: A sample of students aged 14-19 years from the 6 high schools of 1 district of western Turkey were surveyed. The students selected were all attending the school during March and April 2006. The Beck Depression Inventory was used as a screening test. RESULTS: During the study, a total of 846 students completed the survey. Of the study group, 51.9% (439) were male and 48.1% (407) female, with an age average of 16.3 +/- 1.1 years. According to the scale, the prevalence of depression was 30.7% (n = 260), 22.6% for males (n = 99) and 39.6% for females (n = 161). The most depression was seen in males (22.6%), those with any kind of physical problem (37.3%), those with diseases necessitating the use of medication (51.1%), those with acne vulgaris (35.2%), and those having previously experienced any kind of problem (47.3%). CONCLUSIONS: These results highlight not only the need for students' parents and teachers to be well informed on the subject of depression in terms of students' health but also the need for more education programs to be aimed at students relating to the problems they may experience during the period of adolescence. Furthermore, these results show that students identified as depressed should be referred for an appropriate diagnosis to specialized psychiatry centers. PMID- 18387030 TI - An outbreak of itching in an elementary school-a case of mass psychogenic response. PMID- 18387033 TI - Linear spatio-temporal characterization of a UV microscope objective for nonlinear imaging and spectroscopy by using two-dimensional spectral interferometry. AB - Two-dimensional Fourier transform spectral interferometry is used to characterize the spatio-temporal aberrations of a UV microscope objective. The spatial and temporal profiles of a 420 nm, 38 fs pulse at the focus of a 0.32 NA UV objective are then deduced using a wave propagation code incorporating the measured aberrations. PMID- 18387032 TI - In situ detection of programmed cell death in Nicotiana tabacum leaves during senescence. AB - Leaf senescence is a highly regulated developmental process, during which cell constituents are dismantled in an ordered progression. The final death phase of senescence is a type of programmed cell death (PCD). We showed DNA fragmentation characteristic for PCD by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay not only in senescing leaves but also in young, still developing tobacco leaves. PMID- 18387034 TI - Quantifying immunogold labelling in transmission electron microscopy. AB - Gold particles labelling on ultrathin sections is extensively used for antigen localization in transmission electron microscopy. In establishing absolute or relative counts in tissue sections, it would be expedient to use stereologically based unbiased estimates for quantitative results. Nowadays, quantitative immunoelectron microscopy has achieved good and satisfactory results to test whether the gold labelling follows a non-random or a random pattern and then to draw statistical comparisons between cell subcompartments within a sample of cells or between experimental groups of cells. This brief informal review of literature focuses on the relative quantitative determinations of gold labelling of antigens as well as on the statistical distribution comparisons in transmission electron microscopy. PMID- 18387035 TI - Morphological effects of ceramide on DMPC/DHPC bicelles. AB - Freeze fracture electron microscopy and dynamic light scattering were applied to characterize the morphological changes of DMPC/DHPC bicellar systems induced by the addition of ceramides. The results demonstrate a tendency of the DMPC/DHPC aggregates to evolve forming elongated or tubular structures with the increase of the temperature. At 20 degrees C, low concentrations of ceramide promote the appearance of elongated structures with twisted zones. Higher concentrations of this lipid lead to the formation of liposomes along the elongated structures. The increase of the temperature to 40 degrees C induces the growth of the structures containing low concentrations of ceramide forming branched aggregates. In samples with high amounts of ceramide, the increase of temperature causes phase separation and the formation of a mixed system composed by liposomes and multilamellar tubules. The morphological effects induced by ceramides in this new membrane model give new insights for the role played by this lipid in biological membranes. PMID- 18387036 TI - Characterization of the polarization sensitivity anisotropy of a near-field probe using phase measurements. AB - Amplitude and phase measurements of the near-field generated by isolated subwavelength apertures in a gold film are presented. The near-field distribution of such a structure is complex and the measured signal strongly depends on the electric field components effectively detected by the experimental setup. By comparing this signal with 3D vectorial calculations we are able to determine which electric field components are effectively measured. The sensitivity of the phase distribution is key to this measurement. The proposed characterization technique should prove extremely useful to calibrate a Scanning near-field optical microscopy (SNOM) beforehand in order to retrieve quantitative information on the polarization of the field distribution under study. PMID- 18387037 TI - Fabrication of a versatile substrate for finding samples on the nanometer scale. AB - With increasing interest in nanometer scale studies, a common research issue is the need to use different analytical systems with a universal substrate to relocate objects on the nanometer scale. Our paper addresses this need. Using the delicate milling capability of a focused ion beam (FIB) system, a region of interest (ROI) on a sample is labelled via a milled reference grid. FIB technology allows for milling and deposition of material at the sub 20-nm level, in a similar user environment as a standard scanning electron microscope (SEM). Presently commercially available transmission electron microscope (TEM) grids have spacings on the order 100 mum on average; this technique can extend this dimension down to the submicrometre level. With a grid on the order of a few micrometres optical, FIBs, TEMs, scanning electron microscopes (SEMs), and atomic force microscopes (AFM) are able to image the ROI, without special chemical processes or conductive coatings required. To demonstrate, Au nanoparticles of approximately 25 nm in size were placed on a commercial Formvar- and carbon coated TEM grid and later milled with a grid pattern. Demonstration of this technique is also extended to bulk glass substrates for the purpose of sample location. This process is explained and demonstrated using all of the aforementioned analytical techniques. PMID- 18387038 TI - High-pass energy-filtered photoemission electron microscopy imaging of dopants in silicon. AB - Differently doped areas in silicon can show strong electron-optical contrast in dependence on the dopant concentration and surface conditions. Photoemission electron microscopy is a powerful surface-sensitive technique suitable for fast imaging of doping-induced contrast in semiconductors. We report on the observation of Si (100) samples with n- and p-type doped patterns (with the dopant concentration varied from 10(16) to 10(19) cm(-3)) on a p- and n-type substrate (doped to 10(15) cm(-3)), respectively. A high-pass energy filter of the entire image enabled us to obtain spectroscopic information, i.e. quantified photo threshold and related photoyield differences depending on the doping level. Measurements have confirmed the possibility of resolving areas at a high contrast even with the lowest dopant concentration when employing the energy filter. The influence of electron absorption phenomena on contrast formation is discussed. PMID- 18387039 TI - Spatial control of pa-GFP photoactivation in living cells. AB - Photoactivatable green fluorescent protein (paGFP) exhibits peculiar photo physical properties making it an invaluable tool for protein/cell tracking in living cells/organisms. paGFP is normally excited in the violet range (405 nm), with an emission peak centred at 520 nm. Absorption cross-section at 488 nm is low in the not-activated form. However, when irradiated with high-energy fluxes at 405 nm, the protein shows a dramatic change in its absorption spectra becoming efficiently excitable at 488 nm. Confocal microscopes allow to control activation in the focal plane. Unfortunately, irradiation extends to the entire illumination volume, making impracticable to limit the process in the 3D (three-dimensional) space. In order to confine the process, we used two advanced intrinsically 3D confined optical methods, namely: total internal reflection fluorescence (TIRF) and two-photon excitation fluorescence (2PE) microscopy. TIRF allows for spatially selected excitation of fluorescent molecules within a thin region at interfaces, i.e. cellular membranes. Optimization of the TIRF optical set-up allowed us to demonstrate photoactivation of paGFP fused to different membrane localizing proteins. Exploitation of the penetration depth showed that activation is efficiently 3D confined even if limited at the interface. 2PE microscopy overcomes both the extended excitation volume of the confocal case and the TIRF constraint of operating at interfaces, providing optical confinement at any focal plane in the specimen within subfemtoliter volumes. The presented results emphasize how photoactivation by non-linear excitation can provide a tool to increase contrast in widefield and confocal cellular imaging. PMID- 18387040 TI - Navigating transdermal diffusion with multiphoton fluorescence lifetime imaging. AB - We demonstrate the potential of fluorescence lifetime imaging by time-correlated single-photon counting as a method for monitoring the transdermal diffusion pathway and diffusion rate of pharmaceuticals in human skin. The current application relies on observing subtle changes in the fluorescence lifetime of the intrinsic fluorophores present in the intracellular region between corneocytes of the stratum corneum. We have comprehensively characterized the measured fluorescence lifetimes from intracorneocyte junctions in three skin section types (dermatomed skin, epidermal membranes and stratum corneum) revealing statistically significant differences of the short lifetime component between each of the types, which we attribute to the sample preparation and imaging method. We show using epidermal membrane sections that application of a drug/solvent formulation consisting of ethinyl estradiol and spectroscopic grade ethanol to the surface gives rise to a slight but statistically significant shortening of the fluorescence lifetime of the long-lived emitting species present in the sample, from approximately 2.8 ns to 2.5 ns. The method may be useful for future studies where the kinetics and pathways of a variety of applied formulations could be investigated. PMID- 18387041 TI - Starch-based backwards SHG for in situ MEFISTO pulse characterization in multiphoton microscopy. AB - We report a simple methodology to provide complete pulse characterization at the sample plane of a two-photon excited fluorescence (TPEF) microscope. This is achieved by using backward propagating second-harmonic generation (SHG) from starch granules. Without any modification to the microscope, SHG-autocorrelation traces were obtained by using a single starch granule that was placed alongside the biological specimen being imaged. A spectrally resolved SHG autocorrelation was acquired by placing a spectrometer at the output port of the microscope. Complete in situ pulse information is then directly retrieved in an analytical way using the measurement of electric filed by interferometric spectral trace observation (MEFISTO) technique. PMID- 18387042 TI - Two-dimensional dopant profiling with low-energy SEM. AB - The scanning electron microscope has proven itself efficient for determining dopant concentrations in semiconductors. Contrast between differently doped areas is observable in the secondary electron emission. Multiple studies have revealed quantitative relations between the image contrast and dopant concentration. However, intimate examination shows a low reproducibility of the contrast level for a particular local difference between the doping rates. Data about dynamic behaviour of the dopant contrast and its dependence on the status of the sample surface are presented. PMID- 18387043 TI - Study of the structure of the air and blood capillaries of the gas exchange tissue of the avian lung by serial section three-dimensional reconstruction. AB - We have previously reconstructed the gas exchange tissue of the adult muscovy duck, Cairina moschata using a method of manually aligning sections and tracing the contours of the components of the gas exchange tissue. This reconstruction method demonstrated that the air capillaries are comprised of an expanded globular part interconnected by narrow air channels. The blood capillaries completely surround the air capillaries forming an anastomosing meshwork of short segments. However, the resulting reconstruction was limited in scope because of the laborious process of tracing the profiles of each component through the sequence of micrographs. We have now reconstructed a larger proportion of the exchange tissue by using a cross-correlation based alignment strategy and have demonstrated that the staining intensity of each of the exchange tissue components is sufficiently different to allow them to be identified by simple filtering and thresholding. The resulting reconstructions sample a much larger proportion of the exchange tissue and demonstrate the heterogeneity of structures from different locations in the parabronchus. We have shown that a sheet-flow type arrangement of blood capillaries surrounds the infundibulum; this represents an unexpected functional convergence with the arrangement of blood capillaries surrounding the mammalian alveoli. It is feasible, using this reconstruction strategy, to analyse the exchange tissue of a large number of avian species in order to determine structural correlates of function. The resulting reconstructions could be analysed in order to determine the basis of the functional efficiency and rigidity of the avian lung. PMID- 18387045 TI - Dynamic volume change measurements of cereal materials by environmental scanning electron microscopy and videomicroscopy. AB - Moisture migration induced volume change in cereal materials was studied by environmental scanning electron microscopy, videomicroscopy and stereoscopy. It is shown that the in situ volume change can be monitored by the changes of projection area with the assumption of isotropic swelling/shrinkage. The projection area change from videomicroscopy matches well with the volume change by stereoscopy after dimensional normalization. The arbitrary volume values in between two relative humidity levels are available by interpolating the equilibrium volumes at beginning and ending. The sorption/desorption curves were fitted by the 'parallel exponential kinetics' model, which described two independent, parallel processes as 'slow' and 'fast'. In the low relative humidity range, sorption curves match well with the 'parallel exponential kinetics' model and the two parallel processes tend to be identical when relative humidity increases. The diffusivity of water moisture in a cracker, which was estimated by the half-equilibrium principle, has a strong dependence on relative humidity. PMID- 18387044 TI - Deciphering the distribution of organic components in brachiopod shells by confocal laser scanning microscopy. AB - Characterization of the nature and distribution of organic components is crucial to understand shell formation in marine invertebrates. Although several techniques can provide detailed information at high spatial resolution, few of them are non-destructive and informative in a larger structural context. We explore the use of confocal laser scanning microscopy (CLSM) to obtain a better understanding of the distribution of organic components in calcitic shells of brachiopods focusing on perforations (punctae) across the shell. Resulting intensities and patterns of fluorescence correspond well with the distribution of polysaccharides and proteins as reported in previous histological and biochemical studies. Confocal laser microscopy is, therefore, a useful tool to be combined with other techniques to improve our knowledge of biomineral structures in marine invertebrates. PMID- 18387046 TI - Automatic sampling for unbiased and efficient stereological estimation using the proportionator in biological studies. AB - Quantification of tissue properties is improved using the general proportionator sampling and estimation procedure: automatic image analysis and non-uniform sampling with probability proportional to size (PPS). The complete region of interest is partitioned into fields of view, and every field of view is given a weight (the size) proportional to the total amount of requested image analysis features in it. The fields of view sampled with known probabilities proportional to individual weight are the only ones seen by the observer who provides the correct count. Even though the image analysis and feature detection is clearly biased, the estimator is strictly unbiased. The proportionator is compared to the commonly applied sampling technique (systematic uniform random sampling in 2D space or so-called meander sampling) using three biological examples: estimating total number of granule cells in rat cerebellum, total number of orexin positive neurons in transgenic mice brain, and estimating the absolute area and the areal fraction of beta islet cells in dog pancreas. The proportionator was at least eight times more efficient (precision and time combined) than traditional computer controlled sampling. PMID- 18387047 TI - Replicate-based noise corrected correlation for accurate measurements of colocalization. AB - It is widely recognized that the accuracy of colocalization measurements is dependent upon the quality of the source images. We demonstrate that, as the image quality increases, the measured colocalization, using the Pearson and Spearman rank correlation coefficients, approaches the true colocalization asymptotically. This means that in practice it is difficult to obtain images of sufficient quality for accurate measurements. We introduce the replicate-based noise corrected correlation (RBNCC) which aligns the measured colocalization with the true colocalization: a noise measurement is made for each fluorophore from a pair of replicate images, the two noise measurements are combined to generate a correction factor which is applied to the measured colocalization between the two fluorophores. In consequence, accurate measurements can be obtained even with noisy images, making RBNCC especially attractive for live imaging. Even with images of apparently good quality we found an average discrepancy of about 20% between the measured and corrected colocalization. A case is made for using the Spearman rank coefficient instead of the Pearson coefficient to measure colocalization. PMID- 18387048 TI - Upgrading a microscope with a spiral phase plate. AB - We present the implementation of a spiral phase plate in a standard bright-field microscope to enhance the contrast of phase and amplitude samples. The method can be employed in all types of microscopy where standard phase contrast methods are applicable, for example, in bright-field transmission or reflection microscopy using an illumination source with partial spatial coherence. The spiral phase filter is placed into an accessible Fourier plane of the imaging path of the microscope. It is shown that this produces not only a strong contrast enhancement but in theory also improves the spatial resolution of the microscope for white light. A series of different set-ups for transmissive or reflective samples, including epi-illumination, are presented to demonstrate the practical range of applications of this contrasting method. A minute shift of the spiral phase plate out of the centre results in relief-like images that are similar to those obtained by differential interference contrast microscopy. A series of such relief-like images can be numerically processed to obtain quantitative phase and amplitude information of the sample. PMID- 18387049 TI - A statistical approach for intensity loss compensation of confocal microscopy images. AB - In this paper, a probabilistic technique for compensation of intensity loss in confocal microscopy images is presented. For single-colour-labelled specimen, confocal microscopy images are modelled as a mixture of two Gaussian probability distribution functions, one representing the background and another corresponding to the foreground. Images are segmented into foreground and background by applying Expectation Maximization algorithm to the mixture. Final intensity compensation is carried out by scaling and shifting the original intensities with the help of parameters estimated for the foreground. Since foreground is separated to calculate the compensation parameters, the method is effective even when image structure changes from frame to frame. As intensity decay function is not used, complexity associated with estimation of the intensity decay function parameters is eliminated. In addition, images can be compensated out of order, as only information from the reference image is required for the compensation of any image. These properties make our method an ideal tool for intensity compensation of confocal microscopy images that suffer intensity loss due to absorption/scattering of light as well as photobleaching and the image can change structure from optical/temporal section-to-section due to changes in the depth of specimen or due to a live specimen. The proposed method was tested with a number of confocal microscopy image stacks and results are presented to demonstrate the effectiveness of the method. PMID- 18387050 TI - An easily built diffuse illumination system effective at both very low and moderate magnifications, for observing in situ stained slides. AB - Effective study of in situ stained sections often requires illumination that is difficult to achieve with commonly used research microscopes. One must be able to switch quickly and conveniently from the very lowest to moderate magnifications. At all magnifications contrast due to light scatter must be minimized, so that the weak staining that signifies low gene expression can be observed reliably. For the lowest power objectives (e.g., 1.25x or 2x) many microscopes require that the condenser be removed to illuminate the full field of view. This is not only very inconvenient when switching magnifications, but without a condenser the low numerical aperture of the illuminating light beam results in unwanted contrast due to light scatter. We have devised a simple system that diffusely illuminates the full field of view of the lowest power objective (1.25x) and has high enough numerical aperture for use with the 25x and 40x objectives. A key feature is the use of a large diameter ring light (internal diameter 5.8 cm), placed on the microscope base, to illuminate a large diameter diffuser placed just below the slide. PMID- 18387051 TI - The unitary scale bar: human and machine readable. AB - A format is described for a scale bar that encodes the length represented within the structure of the bar itself, thereby removing the need for any supporting text. Although the 'unitary' scale bar has a conventional appearance it is also machine readable and therefore retains information about the scale even when the file format is changed. The format is based on the metre and is suitable for all terrestrial applications. PMID- 18387052 TI - Emerging health economics and outcomes research in the Asia-Pacific region. PMID- 18387053 TI - The association of body mass index with health-related quality of life: an exploratory study in a multiethnic Asian population. AB - OBJECTIVES: To evaluate the association between body mass index (BMI) and health related quality of life (HRQoL) in a multiethnic Asian population in Singapore, and to explore if the World Health Organization (WHO) recommendation of alternative BMI cutoffs for Asians could be further strengthened by evidence of higher risk of impaired HRQoL using these criteria. METHODS: Consenting English, Chinese, Malay and Tamil-speaking primary care patients (age >or= 21 years) were interviewed using English/their respective mother tongue versions of the EQ-5D/EQ VAS, Health Utilities Index (HUI2 & HUI3) and the SF-6D. We first evaluated the relationship between BMI and HRQoL (overall and individual attributes for each instrument) using multiple linear/logistic regression (where appropriate) to adjust for factors known to affect HRQoL. We next reorganized BMI into five categories (reflecting the differences in cutoffs between International/Asian classifications) and evaluated if median HRQoL scores were significantly different across these categories. RESULTS: Among 411 participants [response rate: 87%; median age: 51 years; obese: 19% (International); 33% (Asian)], after adjusting for sociodemographic and other factors, a tendency for underweight and obese subjects to report lower overall HRQoL scores was observed for most instruments. At the individual attribute level, obese subjects reported significantly lower HUI2 pain scores (regression coefficient: -0.035, P = 0.029) and greater odds of reporting problems for SF-6D role-limitations (odds ratio: 2.9, P = 0.005). Median overall HRQoL scores were not significantly different across the five BMI categories. CONCLUSION: Consistent with available studies, obese subjects reported worse HRQoL than normal-weight subjects. That underweight subjects also reported worse HRQoL is interesting and requires confirmation. HRQoL was similar in Asians using either WHO criteria. PMID- 18387054 TI - Economic implications of entecavir treatment in suppressing viral replication in chronic hepatitis B (CHB) patients in China from a perspective of the Chinese Social Security program. AB - OBJECTIVES: Of estimated 112 million persons infected with chronic hepatitis B (CHB) in China, 15% to 40% will eventually develop liver complications. Most patients do not actively seek antiviral agents for treatment due in part to lack of good understanding of the disease. Entecavir is a new therapeutic option for CHB patients and the purpose of this study was to evaluate the cost-effectiveness of entecavir treatment in China, based on projected clinical benefits from its superior viral suppression efficacy. METHODS: The analysis was based on the perspective of the Chinese Social Security program. Adjusted relative risks on the association between viral load (VL) and clinical end points (liver cirrhosis/hepatocellular carcinoma) were derived from a publication of a Taiwan CHB prospective cohort with 42,115 person-years of follow-up, and applied to patients enrolled in a randomized phase III trial in China. In this trial, hepatitis B virus (HBV) DNA (by polymerase chain reaction assay) was the key efficacy end point after 48 weeks of treatment with either entecavir or lamivudine monotherapy. Entecavir and lamivudine daily prices were assumed to be Renminbi Yuan (RMB) 40 and 16.71, respectively. Life expectancy tables were based on China vital statistics. Direct medical cost and utility scores for different phases of CHB were estimated from published China specific data, and costs were adjusted to 2006 values using the Chinese Consumer Price Index. Probabilistic sensitivity analyses were conducted to evaluate parameter uncertainty on event distribution and treatment failure rates beyond the trial period. RESULTS: A total of 519 subjects were enrolled in the study, comprising of 82% males, 87% HBeAg+, and a mean age of 30 years. Based on the efficacy measurement of the percentage of patients achieving HBV DNA <300 copies/ml at week 48, entecavir was superior to lamivudine (78.7% vs. 46.7%, respectively [P < 0.05]). In the base case, compared with lamivudine, 1 year of entecavir therapy gained 0.305 quality adjusted life year (QALY) at an incremental cost of 5368 RMB, with a 3% annual discount. Compared with lamivudine, using entecavir cost an incremental 17,590 RMB per QALY gained (95% CI 6333-56,407). CONCLUSIONS: Based on the results of this study, entecavir is likely to be cost-effective in treating hepatitis B patients in China based on the World Health Organization's recommended maximum willingness to pay threshold. PMID- 18387055 TI - System dynamic modeling: an alternative method for budgeting. AB - OBJECTIVES: To construct, validate, and simulate a system dynamic financial model and compare it against the conventional method. METHODS: The study was a cross sectional analysis of secondary data retrieved from the National Health Security Office (NHSO) in the fiscal year 2004. The sample consisted of all emergency patients who received emergency services outside their registered hospital catchments area. The dependent variable used was the amount of reimbursed money. Two types of model were constructed, namely, the system dynamic model using the STELLA software and the multiple linear regression model. The outputs of both methods were compared. RESULTS: The study covered 284,716 patients from various levels of providers. The system dynamic model had the capability of producing various types of outputs, for example, financial and graphical analyses. For the regression analysis, statistically significant predictors were composed of service types (outpatient or inpatient), operating procedures, length of stay, illness types (accident or not), hospital characteristics, age, and hospital location (adjusted R(2) = 0.74). The total budget arrived at from using the system dynamic model and regression model was US$12,159,614.38 and US$7,301,217.18, respectively, whereas the actual NHSO reimbursement cost was US$12,840,805.69. CONCLUSIONS: The study illustrated that the system dynamic model is a useful financial management tool, although it is not easy to construct. The model is not only more accurate in prediction but is also more capable of analyzing large and complex real-world situations than the conventional method. PMID- 18387056 TI - Issues in drug pricing, reimbursement, and access in China with references to other Asia-Pacific region. AB - OBJECTIVES: Pharmaceutical policies have become paramount in China and other countries of the Asia-Pacific region because of rapidly rising expenditures on drugs. The problems are especially acute in China because expenditures on drugs are typically so large. This article intends to review effects of the policy of drug expenditure containment with primary reference to China, and it proposes some measures to deal with rising pharmaceutical expenditures. METHODS: This article overviews the issues of pharmaceutical pricing, reimbursement, and access in China, and there are a number of policies or measures to control pharmaceutical expenditures. Nevertheless, the effect of those policies of containing drug expenditure is ambiguous so far, and some policies have negative impacts to the manufacturers, providers, and patients. Some underlying reasons are identified. First, the policy's focus on health-care costs is, to some extent, neglected. Second, the governance of the health sector, including pharmaceutical sector, needs to be improved by both the government and the market. RESULTS: This article proposes some suggestions to change policies in drug pricing, reimbursement, and access, and make policies more responsive to the main problem of rising health-care expenditures rather than that of pharmaceutical expenditures alone. CONCLUSIONS: The policy suggestions include those of setting the reasonable price for pharmaceuticals, instituting reasonable incentives for all health decision-makers to encourage efficient use of pharmaceuticals and other health resources, and making pharmaceutical markets more efficient, either in the demand or the supply side. PMID- 18387057 TI - Drug safety surveillance in China and other countries: a review and comparison. AB - OBJECTIVES: Drug safety and postmarketing surveillance have become important public health issues in China. This study reviews the relatively new drug safety surveillance system in China and compares it with the systems in the United States and Europe. METHODS: An extensive literature review was conducted in the following four areas: 1) the organizational structure of the State Food and Drug Administration (SFDA) in China; 2) the development of an adverse drug reaction (ADR) monitoring system in China; 3) regulatory issues related to drug safety in China; and 4) similarities and differences between drug safety surveillance in China and surveillance in the United States and Europe. RESULTS: The SFDA oversees an extensive network of drug safety "watchdogs," including the China National Center for ADR Monitoring and 32 regional centers throughout China. China's system has faced a number of recent challenges. It has had to respond quickly to the withdrawal of various high-profile drugs like Vioxx (rofecoxib) and Baycol (cerivastatin) from other markets. Together with China's Ministry of Health, the SFDA has faced several unique drug safety events. Three of those events, involving the injectable form of the heartleaf houttuyinia herb (Yu Xing Cao), Armillarisni A injections, and clindamycin glucose infusions (Xinfu), are discussed. The rapid development of drug safety surveillance in China is manifested in extensive organizational structure, development of large databases, and laws and regulations supporting drug safety. The two major laws are the China Drug Administration Law issued in February 2001 and the Regulation for the Administration of ADR Reporting and Monitoring issued in March 2004. The study also discusses and compares recent developments in drug safety surveillance in the United States and the European Union. These developments will most likely have implications for the Chinese system in the near future. CONCLUSIONS: While postmarketing surveillance guidelines are not yet available in China, we fully expect their eventual issuance after adaptation to the particular culture and clinical practices in China. PMID- 18387058 TI - Health-care systems and pharmacoeconomic research in Asia-Pacific region. PMID- 18387059 TI - Evidence-based medicine in China. PMID- 18387060 TI - Drug control and formulary management in Malaysia. PMID- 18387061 TI - Health-care delivery system and reimbursement policies in Pakistan. PMID- 18387062 TI - Evidence-based decision-making and health technology assessment in South Korea. PMID- 18387063 TI - Cost-utility analysis of intensive blood glucose control with metformin versus usual care in overweight type 2 diabetes mellitus patients in Beijing, P.R. China. AB - OBJECTIVE: The UKPDS 34 and 51 showed that intensive blood glucose control with metformin is cost-saving and increases life expectancy in overweight type 2 diabetic patients in the United Kingdom. Diabetes is becoming an important health problem in urban China. This study addresses the effects and costs of intensive blood glucose control in this setting, aimed at supporting decision-making on the allocation of scarce resources. METHODS: A decision analytic model was developed to estimate the costs and effectiveness of intensive blood glucose control in overweight type 2 diabetes patients in Beijing, compared with usual care in accordance with clinical practice. The analysis was carried out from a health care perspective. RESULTS: The base-case analysis (3% discount rate) shows that the average incremental costs of 11 years of intensive treatment with metformin are 126.6 K RMB (16.4 K US$) per quality-adjusted life year (QALY) gained. The incremental cost-effectiveness ratio (ICER) is sensitive to the costs of medication alternatives for metformin in the intensive treatment group and to the discount rate used (0%: 105.6 K RMB (13.7 K US$) per QALY gained; 5%: 171.0 K RMB (22.2 K US$) per QALY gained). After 20 and 30 years (lifetime) follow-up, the ICERs become increasingly favorable, 90.1 K RMB (11.7 K US$) and 74.3 K RMB (9.6 K US$), respectively. The ICER is most sensitive to the costs of medication alternatives for metformin in the intensive treatment group, and to the discount rate. CONCLUSIONS: Interpretation of the findings depends on the maximum willingness to pay for a QALY in China, which has not officially been defined. If this would be three times the gross domestic product per capita, a value that has been suggested in the literature, lifetime intensive blood glucose control is likely to be cost-effective. Our findings differ from the UKPDS studies and emphasize that generalizing the results of studies across countries requires considerable adaptation to the local context. PMID- 18387064 TI - Validity, feasibility and acceptability of time trade-off and standard gamble assessments in health valuation studies: a study in a multiethnic Asian population in Singapore. AB - OBJECTIVES: To assess the validity, feasibility and acceptability of standard gamble (SG) and time trade-off (TTO) assessments in a multiethnic Asian population. METHODS: Through in-depth interviews performed among Chinese, Malay, and Indian Singaporeans (education >or= 6 years), we assessed validity of SG/TTO methods for eliciting health preferences by hypothesizing that 1) SG/TTO scores for three hypothetical health states (HS) would exhibit ranked order (decreasing scores with worse HS); and 2) more subjects would rate the most severe HS as worse than dead. Subjects also evaluated feasibility and acceptability of SG/TTO using a 10-point visual analog scale (VAS) and open-ended questions. Ratings were compared using Kruskal-Wallis, Wilcoxon signed-rank tests or tests of proportions. RESULTS: VALIDITY: In 62 subjects (90% response rate), as hypothesized, SG and TTO scores exhibited ranked order with increasing HS severity (SG: 0.85, 0.08, -19.00; TTO: 0.85, 0.00, -0.18). More subjects rated the most severe HS as worse than dead (SG: 8%, 39%, 59%; TTO: 8%, 45% and 62%). FEASIBILITY: Subjects felt SG and TTO were easy to understand (median VAS scores: 8.0 vs. 8.0, P = 0.87) and to complete (8.0 vs. 8.0, P = 0.84). Acceptability: SG and TTO were well accepted, with TTO less so than SG (median [interquartile range] offensiveness: 2.0 [0, 4.0] vs. 2.0 [0, 3.0], P = 0.045). Overall, subjects did not have a clear preference for SG/TTO (50% vs. 45%, P = 0.70). CONCLUSIONS: This study suggests the validity, feasibility and acceptability of SG and TTO for population-based HS valuation studies in a multiethnic Asian population. PMID- 18387065 TI - Cost-effectiveness study comparing cefoperazone-sulbactam to a three-drug combination for treating intraabdominal infections in an Indian health-care setting. AB - OBJECTIVE: This article presents the methodology and results of the pharmacoeconomic analysis of the Magnex Against Standard COmbination Therapy study comparing cefoperazone-sulbactam (Magnex) versus ceftazidime+ amikacin+metronidazole, in the treatment of intra-abdominal infections. METHODS: This prospective, open label, phase IV study was conducted at 17 study sites in India and randomized subjects to receive either cefoperazone-sulbactam or the combination. Pharmacoeconomic analysis was included as a secondary objective and conducted in the clinical efficacy-evaluable (CEE) and the successfully treated patients. All comparisons between treatment groups were conducted using analysis of variance (ANOVA) or Wilcoxon Two-Sample tests. All costs were reported as Indian Rupee (INR) and actual unit costs collected in 2006 were used for the analyses [1 USD approximately 40 INR; 1 Euro approximately 56 INR]. RESULTS: In the CEE and the successfully treated subset of patients, the average cost of treatment was numerically lower in the cefoperazone-sulbactam arm (not statistically significant). The analyses found that the cost-effectiveness ratio (CER) for cefoperazone-sulbactam was INR 17,640.53 and that for the comparator group was INR 22,075.16. Additionally, the incremental CER results showed that the cost of treatment was INR 21,505.59 lower per additional successfully treated patient in the cefoperazone-sulbactam group. CONCLUSIONS: The present study was the first of its kind to be conducted in the "price sensitive" Indian health-care setting. Though study was not powered for the difference in average cost of treatments, there was a trend favoring cefoperazone sulbactam. The findings from this study should encourage further conduct of similar analyses and increase the knowledge regarding pharmacoeconomics in India. PMID- 18387066 TI - Comparison of the initial hospitalization costs between the patients treated with dobutamine and the patients treated with amrinone for acute decompensated heart failure in a Japanese institute. AB - OBJECTIVES: Phosphodiesterase (PDE) III inhibitor therapy is effective for treatment of acute decompensated heart failure (ADHF). Nevertheless, this drug is expensive than conventional inotropic agent dobutamine. We compared total medication costs of the patients treated with PDE III inhibitor amrinone therapy to that of the patients treated with conventional dobutamine therapy during initial hospitalization. METHODS: We analyzed 160 consecutive patients with ADHF admitted to our hospital. Shock, dehydration, severe infection, multiple organ failure, and mild heart failure (New York Heart Association class IIs) were not eligible for the study. Ninety-seven patients were divided into two groups: 1) DOB group treated with dobutamine therapy; and 2) AMR group treated with amrinone therapy. Total medication costs and cost for hospital room charge were calculated based on their usage during the initial hospitalization for each patient. Group comparison was done between the DOB and AMR groups. RESULTS: Length of stay was longer in the DOB group than in the AMR group. Mean calculated cost of intravenous drugs was higher in the DOB group (173,186 +/- 239,147 yen) than in the AMR group (63,145 +/- 47,223 yen, P < 0.05). Total medication costs were higher in the DOB group than in the AMR group. Cost for hospital room charge was higher in the DOB group than in the AMR group. CONCLUSIONS: In the treatment of ADHF, appropriate therapy even with expensive drugs makes total medication costs less expensive comparing with conventional therapy with cheaper drugs during initial hospitalization. PMID- 18387067 TI - Cost-effectiveness analysis of thiazolidinediones in uncontrolled type 2 diabetic patients receiving sulfonylureas and metformin in Thailand. AB - OBJECTIVE: The national essential drug committee in Thailand suggested that only one of thiazolidinediones be included in hospital formulary but little was know about their cost-effectiveness values. This study aims to determine an incremental cost-effectiveness ratio of pioglitazone 45 mg compared with rosiglitazone 8 mg in uncontrolled type 2 diabetic patients receiving sulfonylureas and metformin in Thailand. METHODS: A Markov diabetes model (Center for Outcome Research model) was used in this study. Baseline characteristics of patients were based on Thai diabetes registry project. Costs of diabetes were calculated mainly from Buddhachinaraj hospital. Nonspecific mortality rate and transition probabilities of death from renal replacement therapy were obtained from Thai sources. Clinical effectiveness of thiazolidinediones was retrieved from a meta-analysis. All analyses were based on the government hospital policymaker perspective. Both cost and outcomes were discounted with the rate of 3%. Base-case analyses were analyzed as incremental cost per quality-adjusted life year (QALY) gained. A series of sensitive analyses were performed. RESULTS: In base-case analysis, the pioglitazone group had a better clinical outcomes and higher lifetime costs. The incremental cost per QALY gained was 186,246 baht (US$ 5389). The acceptability curves showed that the probability of pioglitazone being cost-effective was 29% at the willingness to pay of one time of Thai gross domestic product per capita (GDP per capita). The effect of pioglitazone on %HbA1c decrease was the most sensitive to the final outcomes. CONCLUSIONS: Our findings showed that in type 2 diabetic patients who cannot control their blood glucose under the combination of sulfonylurea and metformin, the use of pioglitazone 45 mg fell in the cost-effective range recommended by World Health Organization (one to three times of GDP per capita) on average, compared to rosiglitazone 8 mg. Nevertheless, based on sensitivity analysis, its probability of being cost-effective was quite low. Hospital policymakers may consider our findings as part of information for the decision-making process. PMID- 18387068 TI - A difficult balancing act: policy actors' perspectives on using economic evaluation to inform health-care coverage decisions under the Universal Health Insurance Coverage scheme in Thailand. AB - OBJECTIVES: In Thailand, policymakers have come under increasing pressure to use economic evaluation to inform health-care resource allocation decisions, especially after the introduction of the Universal Health Insurance Coverage (UC) scheme. This article presents qualitative findings from research that assessed a range of policymakers' perspectives on the acceptability of using economic evaluation for the development of health-care benefit packages in Thailand. The policy analysis examined their opinions about existing decision-making processes for including health interventions in the UC benefit package, their understanding of health economic evaluation, and their attitudes, acceptance, and values relating to the use of the method. METHODS: Semistructured interviews were conducted with 36 policy actors who play a major role or have some input into health resource allocation decisions within the Thai health-care system. These included 14 senior policymakers at the national level, 5 hospital directors, 10 health professionals, and 7 academics. RESULTS AND CONCLUSIONS: Policy actors thought that economic evaluation information was relevant for decision-making because of the increasing need for rationing and more transparent criteria for making UC coverage decisions. Nevertheless, they raised several difficulties with using economic evaluation that would pose barriers to its introduction, including distrust in the method, conflicting philosophical positions and priorities compared to that of "health maximization," organizational allegiances, existing decision-making procedures that would be hard to change, and concerns about political pressure and acceptability. PMID- 18387069 TI - Applying the Expectancy-Value Model to understand health values. AB - OBJECTIVES: Expectancy-Value Model (EVM) is the most structured model in psychology to predict attitudes by measuring attitudinal attributes (AAs) and relevant external variables. Because health value could be categorized as attitude, we aimed to apply EVM to explore its usefulness in explaining variances in health values and investigate underlying factors. METHODS: Focus group discussion was carried out to identify the most common and significant AAs toward 5 different health states (coded as 11111, 11121, 21221, 32323, and 33333 in EuroQol Five-Dimension (EQ-5D) descriptive system). AAs were measured in a sum of multiplications of subjective probability (expectancy) and perceived value of attributes with 7-point Likert scales. Health values were measured using visual analog scales (VAS, range 0-1). External variables (age, sex, ethnicity, education, housing, marital status, and concurrent chronic diseases) were also incorporated into survey questionnaire distributed by convenience sampling among eligible respondents. Univariate analyses were used to identify external variables causing significant differences in VAS. Multiple linear regression model (MLR) and hierarchical regression model were used to investigate the explanatory power of AAs and possible significant external variable(s) separately or in combination, for each individual health state and a mixed scenario of five states, respectively. RESULTS: Four AAs were identified, namely, "worsening your quality of life in terms of health" (WQoL), "adding a burden to your family" (BTF), "making you less independent" (MLI) and "unable to work or study" (UWS). Data were analyzed based on 232 respondents (mean [SD] age: 27.7 [15.07] years, 49.1% female). Health values varied significantly across 5 health states, ranging from 0.12 (33333) to 0.97 (11111). With no significant external variables identified, EVM explained up to 62% of the variances in health values across 5 health states. The explanatory power of 4 AAs were found to be between 13% and 28% in separate MLR models (P < 0.05). When data were analyzed for each health state, variances in health values became small and explanatory power of EVM was reduced to a range between 8% and 23%. CONCLUSION: EVM was useful in explaining variances of health values and predicting important factors. Its power to explain small variances might be restricted due to limitations of 7-point Likert scale to measure AAs accurately. With further improvement and validation of a compatible continuous scale for more accurate measurement, EVM is expected to explain health values to a larger extent. PMID- 18387070 TI - Factors affecting health-care costs and hospitalizations among diabetic patients in Thai public hospitals. AB - OBJECTIVE: The study investigated the factors affecting health-care costs and hospitalizations among diabetic patients in Thai public hospitals. METHODS: A retrospective study was conducted by using administrative claims data obtained from diabetic patients during October 1, 2002 and September 30, 2003. Dependent variables were total health-care costs and the occurrence of hospitalizations. Independent variables included demographic factors, health-care utilisation, complications, comorbidities, and payment methods. Multivariate statistical analyses were applied. RESULTS: The results of this study suggested that demographic factors of patients (i.e., age and male sex), payment methods (i.e., capitation, fee-for-service, and out-of-pocket) were significantly associated with higher health-care costs and probability of hospitalization. Patients receiving treatment from teaching hospitals significantly consumed higher health care costs. In addition, the more health-care utilisation (i.e., occurrence of hospitalization, number of outpatient visit, and insulin utilization), the higher health-care costs the patients significantly had. Diabetic patients taking insulin had significantly higher health-care costs and risk of hospitalization. Furthermore, comorbidities (e.g., hypertension and cancer) and diabetes-related complications (e.g., nephropathy, neuropathy, retinopathy, coronary artery disease, cardiovascular disease, and peripheral vascular disease) were significantly associated with an increase in health-care costs and hospitalization. CONCLUSION: Factors affecting health-care costs and hospitalizations may help health-care providers intervene to improve patient management and possibly reduce health-care costs in the future. PMID- 18387071 TI - A cost function analysis of shigellosis in Thailand. AB - OBJECTIVE: The purpose of this study was to develop a cost function model to estimate the public treatment cost of shigellosis patients in Thailand. METHODS: This study is an incidence-based cost-of-illness analysis from a provider's perspective. The sample cases in this study were shigellosis patients residing in Kaengkhoi District, Saraburi Province, Thailand. All diarrhea patients who came to the health-care centers in Kaengkhoi District, Kaengkhoi District Hospital and Saraburi Regional Hospital during the period covering May 2002 to April 2003 were tested for Shigella spp. The sample for our study included all patients with culture that confirmed the presence of shigellosis. Public treatment cost was defined as the costs incurred by the health-care service facilities arising from individual cases. The cost was calculated based on the number of services that were utilized (clinic visits, hospitalization, pharmaceuticals, and laboratory investigations), as well as the unit cost of the services (material, labor and capital costs). The data were summarized using descriptive statistics. Furthermore, the stepwise multiple regressions were employed to create a cost function, and the uncertainty was tested by a one-way sensitivity analysis of varying discount rate, cost category, and drug prices. RESULTS: Cost estimates were based from 137 episodes of 130 patients. Ninety-four percent of them received treatment as outpatients. One-fifth of the episodes were children aged less than 5 years old. The average public treatment cost was US$8.65 per episode based on 2006 prices (95% CI, 4.79, and 12.51) (approximately US$1 = 38.084 Thai baht). The majority of the treatment cost (59.3%) was consumed by the hospitalized patients, though they only accounted for 5.8% of all episodes. The sensitivity analysis on the component of costs and drug prices showed a variation in the public treatment cost ranging from US$8.29 to US$9.38 (-4.20% and 8.43% of the base-case, respectively). The public treatment cost model has an adjusted R(2) of 0.788. The positive predictor variables were types of services (inpatient and outpatient), types of health-care facilities (health center, district hospital, regional hospital), and insurance schemes (civil servants medical benefit scheme, social security scheme and universal health coverage scheme). Treatment cost was estimated for various scenarios based on the fitted cost model. CONCLUSION: The average public treatment cost of shigellosis in Thailand was estimated in this study. Service types, health-care facilities, and insurance schemes were the predictors used to predict nearly 80% of the cost. The estimated cost based on the fitted model can be employed for hospital management and health care planning. PMID- 18387072 TI - A study on indirect and intangible costs for patients with knee osteoarthritis in Singapore. AB - OBJECTIVES: To estimate indirect costs through human capital approach and intangible costs through willingness-to-pay (WTP), and identify factors potentially affecting these costs in multiethnic Asian patients with knee osteoarthritis (OA). METHODS: Data were collected through face-to-face interviews among knee OA patients. Human capital approach was used to estimate indirect costs by multiplying: 1) days of absence from work because of OA, with average earnings per capita per day for working patients; or 2) productivity loss with the market price of housekeeping for retirees/homemakers. A closed-ended iterative bidding contingent valuation method was used to elicit willingness-to pay for a hypothetical cure of OA as a proxy for intangible costs. Mann-Whitney U or Kruskal-Wallis H-tests were performed in univariate analyzes, and linear regression in multivariate analyses. RESULTS: Indirect costs per year and intangible costs were estimated at US$1008 and US$1200, accounting for 2.8% and 3.3% of annual household income, respectively. The indirect costs were significantly higher for male or working patients, while intangible costs were higher for Chinese, working patients, with higher income, or worse global well being. CONCLUSION: This study demonstrated that eliciting indirect costs through human capital approach and intangible costs through WTP are acceptable and feasible in Asian patients with knee OA. Besides the direct costs, the indirect and intangible costs for the OA patients could be substantial. PMID- 18387073 TI - Cholesterol goal attainment in patients with coronary heart disease and elevated coronary risk: results of the Hong Kong hospital audit study. AB - OBJECTIVE: We sought to determine 1) long-term lipid-lowering treatment patterns; 2) cholesterol goal attainment rates and possible determinants of goal achievement; and 3) effects of cholesterol goal attainment on coronary events in hospitalized Hong Kong patients. METHODS: In this retrospective cohort analysis, records of two public Hong Kong hospitals were reviewed for 196 adults (69% with coronary heart disease (CHD) or CHD-risk equivalent) who received at least one lipid-lowering therapy during hospitalization. Low-density lipoprotein cholesterol (LDL-C) targets were <2.6 mmol/l (<100 mg/dL) for patients with CHD or CHD risk equivalents and <3.37 mmol/l (<130 mg/dL) for those without. RESULTS: Most participants were initiated on regimens of low to midequipotency doses and never had their regimens adjusted to higher potency. Approximately 44% of patients not at LDL-C at baseline failed to achieve goal during a median follow up of 1.9 years. Patients with higher coronary risk and/or LDL-C levels at baseline were less likely than their lower-risk counterparts to achieve goal; for each 1-mmol/l (38.7-mg/dL) increase in LDL-C at baseline, the likelihood of attaining goal declined by 64%. Patients achieving cholesterol goal had significantly longer cardiovascular event-free times. CONCLUSIONS: A total of 44% of Hong Kong patients not at LDL-C goals at baseline did not achieve them over 1.9 years. More effective and well-tolerated therapies, including adjunctive regimens (e.g., ezetimibe-statin, niacin-statin), may be necessary to enhance LDL C goal achievement and increase event-free time. PMID- 18387074 TI - The reliability and validity of a Korean translation of the BASDAI in Korean patients with ankylosing spondylitis. AB - OBJECTIVES: The objective of this study was to develop a Korean version of the original English version of BASDAI (Bath Ankylosing Spondylitis Disease Activity Index) and to evaluate its reliability and validity in Korean patients with Ankylosing Spondylitis (AS). METHODS: A total of 50 outpatients diagnosed as AS by the modified New York criteria participated. To develop a Korean version of the BASDAI, we followed rigorous international translation steps and evaluated reliability and validity by calculating Cronbach's alpha and correlation coefficients between BASDAI score and clinical parameters (e.g., ESR, CRP, modified Schober test index, finger-to-ground index, and Bath Ankylosing Spondylitis Radiologic Index Score, and bone mineral density). RESULTS: Cronbach's alpha (= 0.75) was acceptable. The distribution of item responses evaluated by the ceiling and floor effects showed appropriate proportions and a good discrimination with the Korean version of the BASDAI. The correlations among the mean BASDAI score and five scales for the convergent validity was significantly correlated with each other (all P-values < 0.01). The correlations between the BASDAI score and both erythrocyte sedimentation rate and C-reactive protein for the criterion validity were positively correlated (all P-values < 0.05). The results of this study showed that the Korean translation of the BASDAI is an efficient tool in terms of its reliability and validity for the measurement of the disease activity in patients with AS. CONCLUSIONS: The Korean version of the BASDAI could be used in clinical research to assess and evaluate the course of disease activity in Korean AS patients. PMID- 18387075 TI - Biotic interactions and macroevolution: extensions and mismatches across scales and levels. AB - Clade dynamics in the fossil record broadly fit expectations from the operation of competition, predation, and mutualism, but data from both modern and ancient systems suggest mismatches across scales and levels. Indirect effects, as when antagonistic or mutualistic interactions restrict geographic range and thereby elevate extinction risk, are probably widespread and may flow in both directions, as when species- or organismic-level factors increase extinction risk or speciation probabilities. Apparent contradictions across scales and levels have been neglected, including (1) the individualistic geographic shifts of species on centennial and millennial timescales versus evidence for fine-tuned coevolutionary relationships; (2) the extensive and dynamic networks of interactions faced by most species versus the evolution of costly enemy-specific defenses and finely attuned mutualisms; and (3) the macroevolutionary lags often seen between the origin and the diversification of a clade or an evolutionary novelty versus the rapid microevolution of advantageous phenotypes and the invasibility of most communities. Resolution of these and other cross-level tensions presumably hinges on how organismic interactions impinge on genetic population structures, geographic ranges, and the persistence of incipient species, but generalizations are not yet possible. Paleontological and neontological data are both incomplete and so the most powerful response to these problems will require novel integrative approaches. Promising research areas include more realistic approaches to modeling and empirical analysis of large scale diversity dynamics of ostensibly competing clades; spatial and phylogenetic dissections of clades involved in escalatory dynamics (where prey respond evolutionarily to a broad and shifting array of enemies); analyses of the short- versus long-term consequences of mutualistic symbioses; and fuller use of abundant natural experiments on the evolutionary impacts of ecosystem engineers. PMID- 18387076 TI - Consensus meeting on reporting glycated haemoglobin (HbA1c) and estimated average glucose (eAG) in the UK: report to the National Director for Diabetes, Department of Health. PMID- 18387077 TI - Matrix metalloproteinases and diabetic foot ulcers: the ratio of MMP-1 to TIMP-1 is a predictor of wound healing. AB - AIMS: Matrix metalloproteinases (MMPs) play a major role in wound healing: they can degrade all components of the extracellular matrix. In diabetic foot ulcers there is an excess of MMPs and a decrease of the tissue inhibitors of MMPs (TIMPs). This imbalance is probably one cause of impaired healing. However, little is known about changes in MMPs during wound healing. METHODS: Sixteen patients with neuropathic diabetic foot ulcers participated. Wound fluid was collected regularly during the 12-week follow-up period, for measurement of MMP 1, MMP-2, MMP-8, MMP-9 and TIMP-1. Results were analysed by the degree of wound healing: good healers (defined by a reduction of at least 82% in initial wound surface at 4 weeks) and poor healers (reduction of less than 82% in wound surface at 4 weeks). RESULTS: In good healers, levels of MMP-8 and -9 secreted by inflammatory cells decreased earlier. The initial levels of MMP-1 were similar in good and poor healers (P = 0.1) but rose significantly at week 2 in good healers (P = 0.039). There was a significant correlation between a high ratio of MMP 1/TIMP-1 and good healing (r = 0.65, P = 0.008). Receiver Operator Curve (ROC) analysis showed that an MMP-1/TIMP-1 ratio of 0.39 best predicted wound healing (sensitivity = 71%, specificity = 87.5%). CONCLUSIONS: A high level of MMP-1 seems essential to wound healing, while an excess of MMP-8 and -9 is deleterious, and could be a target for new topical treatments. The MMP-1/TIMP-1 ratio is a predictor of wound healing in diabetic foot ulcers. PMID- 18387078 TI - Long-term efficacy and safety of insulin detemir compared to Neutral Protamine Hagedorn insulin in patients with Type 1 diabetes using a treat-to-target basal bolus regimen with insulin aspart at meals: a 2-year, randomized, controlled trial. AB - AIMS: This 24-month, multi-national, open-label, parallel group trial investigated the long-term efficacy and safety of insulin detemir and Neutral Protamine Hagedorn insulin in combination with mealtime insulin aspart in patients with Type 1 diabetes using a treat-to-target concept. METHODS: Patients were randomized 2 : 1 to detemir (n = 331) or NPH (n = 166) groups. Basal insulin was initiated once daily (evening) and titrated individually based on self measured plasma glucose (PG) levels, aiming for pre-breakfast and pre-dinner targets < or = 6.0 mmol/l. A second basal morning dose could be added according to pre-defined criteria. RESULTS: After 24 months, superiority of glycated haemoglobin (HbA(1c)) was achieved with detemir compared to NPH (detemir 7.36%, NPH 7.58%, mean difference -0.22% points) [95% confidence interval (CI) -0.41 to 0.03%], with reductions of 0.94% and 0.72% points, respectively. Fasting PG (FPG(lab)) was also lower with detemir (detemir 8.35 mmol/l, NPH 9.43 mmol/l; P = 0.019). Twenty-two per cent of patients treated with detemir reached an HbA(1c) < or = 7.0% in the absence of confirmed hypoglycaemia during the last month of treatment vs. 13% on NPH (P = 0.019). Risk of major and nocturnal hypoglycaemia was 69% and 46% lower with detemir than with NPH (P < 0.001), respectively; patients treated with detemir gained less weight (detemir 1.7 kg, NPH 2.7 kg; P = 0.024). The overall safety profile was similar in the two groups and treatment with detemir did not result in any unexpected findings. CONCLUSIONS: Long-term treatment with the insulin analogues detemir + aspart was superior to NPH + aspart in reducing HbA(1c), with added benefits of less major and nocturnal hypoglycaemia and less weight gain. PMID- 18387079 TI - Continuous Subcutaneous Glucose Monitoring System in diabetic mothers during labour and postnatal glucose adaptation of their infants. AB - AIMS: To assess a new technique for continuous monitoring of glucose concentration during labour in diabetic mothers. A second objective was to study maternal glucose levels in relation to postnatal glucose adaptation and the need for intravenous (IV) glucose treatment in the newborn infant. METHODS: Fifteen pregnant women with insulin-treated diabetes mellitus participated in this prospective pilot study. To measure their glucose control during labour we used the Continuous Subcutaneous Glucose Monitoring System (CGMS; Medtronic, Minneapolis, MN, USA) to calculate the mean glucose concentration and the area under the curve (AUC) in the last 120 min before delivery. All infants of these women were transferred to the neonatal care unit for early oral feeding and blood glucose measurements up to 14 h after delivery. Infants received IV glucose if blood glucose values were repeatedly < 2.2 mmol/l. RESULTS: All women coped well with the CGMS monitoring. AUC 0-120 min before delivery, mean glucose concentration 0-120 min before delivery and cord plasma insulin level were all significantly associated with the need for IV glucose in the newborn children. CONCLUSIONS: In this study we found an association between maternal glucose concentrations during labour and postnatal glucose adaptation and need for IV glucose treatment in the infants. Online monitoring of glucose levels during delivery might help us to achieve maternal normoglycaemia and further reduce the risk of postnatal hypoglycaemia in the offspring. PMID- 18387080 TI - Prevalence of impaired awareness of hypoglycaemia in adults with Type 1 diabetes. AB - AIMS: Impaired awareness of hypoglycaemia (IAH) is thought to affect approximately 25% of people with Type 1 diabetes. While this estimate was based on retrospective information from patients in several small studies performed several years ago, validated methods of assessment have not been used in a large hospital clinic-based population to ascertain the prevalence in the present era. METHODS: Five hundred and eighteen people with Type 1 diabetes were recruited by random selection over a 2-year period. Participants completed a questionnaire documenting baseline characteristics and assessment of their awareness status using the method described by Gold et al. The number of episodes of severe hypoglycaemia they had experienced in the preceding year was recorded retrospectively. RESULTS: IAH was present in 19.5% of the cohort. Compared to those with normal awareness of hypoglycaemia, those with IAH were significantly older [mean +/- standard deviation (sd); 39.3 +/- 12.9 vs. 45.9 +/- 13.5 years, P < 0.001], had a longer duration of diabetes [median (interquartile range) 14 (8 22) vs. 23 (14-32) years, P < 0.001], and had a six-fold higher frequency of severe hypoglycaemia in the previous year [0.38 +/- 1.04 (25th-75th centile 0-0) vs. 2.36 +/- 4.81 (25th-75th centile 0-2) episodes per person, P < 0.001]. CONCLUSIONS: The present survey of a large hospital-based clinic population has confirmed that a significant proportion of people with Type 1 diabetes (19.5%) continue to have IAH. Despite improvements in insulin therapies, intensification of insulin regimens and innovative patient education, the prevalence of IAH remains high in Type 1 diabetes. PMID- 18387081 TI - Ethnic differences in plantar pressures in diabetic patients with peripheral neuropathy. AB - AIMS: To compare plantar foot pressures between Caucasian and Hispanic diabetic patients with peripheral neuropathy (PN) without a history of foot ulceration and between Caucasian and Hispanic non-diabetic individuals. METHODS: Forty-four Hispanic diabetic patients with PN (HDPN), 35 Caucasian diabetic patients with PN (CDPN), 41 non-diabetic Hispanic subjects and 33 non-diabetic Caucasian subjects participated. Total and regional peak plantar pressures (PPs) and pressure time integrals (PTIs) were assessed using the EMED-SF-4 plantar pressure system. RESULTS: Hispanic diabetic patients with PN had significantly lower peak PP than Caucasian diabetic patients with PN in the entire foot (552.4 +/- 227.9 vs. 810.1 +/- 274.6 kPa; P < 0.001), forefoot (464.1 +/- 222.6 vs. 699.6 +/- 323.1 kPa; P < 0.001), hindfoot (296.3.4 + 101.8 vs. 398.1 + 178.3 kPa; P < 0.01) and at the fifth metatarsal head (MTH5; 204.3 +/- 143.2 vs. 388.2 +/- 273.9 kPa; P < 0.001). The PTI in the entire foot, forefoot and MTH5 were also lower in HDPN than in CDPN. The ethnic differences between the diabetic groups with PN for the entire foot, forefoot and MTH5 remained significant after adjusting for the effect of age, gender, weight and duration of diabetes. There were no significant differences in peak PP and PTI among non-diabetic individuals, except for a lower peak PP at the MTH5 in Hispanic compared with Caucasian subjects. CONCLUSIONS: Despite a well-known higher incidence of foot complications in diabetic Hispanic subjects, dynamic plantar pressures are lower in Hispanic diabetic patients with PN when compared with their Caucasian counterparts, suggesting that differences in other risk factors exist between these two ethnic groups. PMID- 18387083 TI - Quantification of cardiac fibrosis by colour-subtractive computer-assisted image analysis. AB - 1. Quantification of fibrosis is a key parameter in the assessment of the severity of cardiovascular disease and efficacy of future candidate therapies. Computer-assisted methods are frequently used to assess cardiac fibrosis in several experimental models. A brief survey indicated that there is a clear dearth of literature outlining detailed methodologies for computer-based assessment of cardiac fibrosis. The purpose of the present study was to provide a reliable method for a systematic assessment of cardiac fibrosis. 2. We induced cardiac fibrosis by isoproterenol (ISO) infusion in adult CD1 male mice and quantified fibrosis using a recently developed colour-subtractive computer assisted image analysis (CS-CAIA) technique. Here, we provided a detailed description of our methodology to facilitate its wider use by other researchers. 3. We showed that the severity of ISO-induced cardiac fibrosis was similar in the apex, mid-ventricular ring and base of the adult CD1 mouse heart. In contrast with other species, such as rats and dogs, we found that uniform expression of beta(1)-adrenoceptors between different regions in CD1 mouse hearts correlated well with uniform induction of cardiac fibrosis. 4. A previous study found a negative correlation between levels of myocardial fibrosis and the degree of cardiac hypertrophy in ISO-treated Wistar rats. In contrast, we found a similar degree of cardiac fibrosis in our ISO-treated CD1 mice. 5. Our results suggest that CD1 mice are an ideal model system to study catecholamine-induced cardiac remodelling, as well as to screen candidate antifibrotic agents for future therapies. PMID- 18387086 TI - A study of the soluble complexes formed during calcium binding by soybean protein hydrolysates. AB - The soluble complexes formed between hydrolyzed soybean protein and calcium at pH 7.4 were investigated using dialysis, gel chromatography, and Fourier transform infrared spectrometry (FTIR). The results demonstrate that the amount of calcium bound was significantly different for soybean protein hydrolysates obtained using the proteases neutrase, flavourzyme, protease M, and pepsin. Maximum levels of calcium binding (66.9 mg/g) occurred with hydrolysates produced using protease M. Peptide fragments exhibiting high calcium binding capacity had molecular weights of either 14.4 kDa or 8 to 9 kDa, and the calcium binding capacity was linearly correlated with carboxyl group content (R(2)= 0.8204). FTIR experiments revealed that upon binding calcium, the amide I band underwent a shift to lower wave numbers. A wide, intense Ca-O absorption band also appeared between 400 and 100 cm(-1) in the far-infrared spectrum. The width and intensity of this band increased after treatment of samples with glutaminase. The amount of bound calcium was related to both the molecular weight of the peptides and to the carboxyl group content, and the most likely sites for calcium binding are the carboxyl groups of Asp and Glu. PMID- 18387084 TI - A role for benzamil-sensitive proteins of the central nervous system in the pathogenesis of salt-dependent hypertension. AB - 1. Although increasing evidence suggests that salt-sensitive hypertension is a disorder of the central nervous system (CNS), little is known about the critical proteins (e.g. ion channels or exchangers) that play a role in the pathogenesis of the disease. 2. Central pathways involved in the regulation of arterial pressure have been investigated. In addition, systems such as the renin angiotensin-aldosterone axis, initially characterized in the periphery, are present in the CNS and seem to play a role in the regulation of arterial pressure. 3. Central administration of amiloride, or its analogue benzamil hydrochloride, has been shown to attenuate several forms of salt-sensitive hypertension. In addition, intracerebroventricular (i.c.v.) benzamil effectively blocks pressor responses to acute osmotic stimuli, such as i.c.v. hypertonic saline. Amiloride or its analogues have been shown to interact with the brain renin-angiotensin-aldosterone system (RAAS) and to effect the expression of endogenous ouabain-like compounds. Alterations of brain RAAS function and/or endobain expression could play a role in the interaction between amiloride compounds and arterial pressure. Peripheral treatments with benzamil, even at higher doses than those given centrally, have little or no effect on arterial pressure. These data provide strong evidence that benzamil-sensitive proteins (BSPs) of the CNS play a role in cardiovascular responsiveness to sodium. 4. Mineralocorticoids have been linked to human hypertension; many patients with essential hypertension respond well to pharmacological agents antagonizing the mineralocorticoid receptor and certain genetic forms of hypertension are caused by chronically elevated levels of aldosterone. The deoxycorticosterone acetate (DOCA)-salt model of hypertension is a benzamil-sensitive model that incorporates several factors implicated in the aetiology of human disease, including mineralocorticoid action and increased dietary sodium. The DOCA-salt model is ideal for investigating the role of BSPs in the pathogenesis of hypertension, because mineralocorticoid action has been shown to modulate the activity of at least one benzamil-sensitive protein, namely the epithelial sodium channel. 5. Characterizing the BSPs involved in the pathogenesis of hypertension may provide a novel clinical target. Further studies are necessary to determine which BSPs are involved and where, in the nervous system, they are located. PMID- 18387087 TI - Effect of preservative agents on the respiration rate of minimally processed potato (Solanum tuberosum cv. Monalisa). AB - The shelf life of minimally processed potatoes (MPP) is limited by enzyme catalyzed browning reactions, with the increase in respiration being another factor that affects quality retention of this product. Sulfites are commonly used as effective preservative agents in minimally processing potatoes, but ascorbic acid and citric acid are considered natural sulfite substitutes and more accepted by consumers. The aim of this study was to study the effect of combinations of the preservative agents cited above (sodium metabisulfite 0.1% and 0.5%; citric acid 0.1% and 0.5%; ascorbic acid 0.5%) on the respiration rate of MPP (cv. Monalisa) processed at both ambient and refrigerated temperatures. The results have revealed that there is a significant effect of dipping treatment and temperature on respiration rate of MPP. Sodium metabisulfite (SM) reduces respiratory activity up to 0.8 mL/kg/h. The addition of either citric or ascorbic acid enhanced the effect of SM on the reduction of the respiration rate of MPP. The strongest effect (up to 3.3 mL/kg/h) was observed when a combination of all 3 agents at the higher concentrations was employed at a temperature of 18 degrees C. PMID- 18387088 TI - Effects of pork meat cut and packaging type on lipid oxidation and oxidative products during refrigerated storage (8 degrees C). AB - Lipid oxidation and oxidative products as affected by pork meat cut, packaging method, and storage time were evaluated during refrigerated storage. Pork belly had higher pH and thiobarbituric acid reactive substance (TBARS) values than pork loin, and aerobic-packaged belly had higher TBARS than vacuum-packaged counterparts. Loin had higher free fatty acid (FFA) values than belly, and increased FFA values were observed with increased storage time. Peroxide values increased up to 7 d and decreased thereafter. Volatile compounds such as alkanes, aldehydes, ketones, and alcohols with high volatility in belly were higher than those in loin. Nonanoic acid, ethyl ester in belly, and hexadecanoic acid in loin might be considered as indices of lipid oxidation. Overall, vacuum packaging was better than aerobic packaging to retard lipid oxidation and production of oxidative products, and loin was more sensitive to lipid oxidation than belly. PMID- 18387089 TI - Study of the biosynthesis of 1-octen-3-ol using a crude homogenate of Agaricus bisporus in a bioreactor. AB - 1-Octen-3-ol and 10-oxo-trans-8-decenoic acid are metabolites of the breakdown of linoleic acid (LA) by mushroom enzymes. These compounds can be produced in a bioreactor using a crude mushroom homogenate and the exogenous addition of LA and oxygen. The factors' duration of blending, mushroom-buffer ratio, effect of a surfactant, whole against partially clarified reaction broths, purity of LA, and utilization of stumps instead of whole mushrooms were studied for their effect on reaction yield using a 1-L bioreactor. The results showed the feasibility of using the more inexpensive 60%-pure LA instead of the 99%-pure LA even when a yield loss was involved. Waste stumps could be used instead of whole mushrooms with a yield decline of 26%. PMID- 18387090 TI - Properties of high-oleic palm oils derived by fractional crystallization. AB - High-oleic palm oil (HOPO) with an oleic acid content of 59.0% and an iodine value (IV) of 78.2 was crystallized in a 200-kg De Smet crystallizer with a predetermined cooling program and appropriate agitation. The slurry was then fractionated by means of dry fractionation at 4, 8, 10, 12, and 15 degrees C. The oil and the fractionated products were subjected to physical and chemical analyses, including fatty acid composition, triacylglycerol and diacylglycerol composition, solid fat content, cloud point, slip melting point, and cold stability test. Fractionation at 15 degrees C resulted in the highest olein yield but with minimal oleic acid content. Due to the enhanced unsaturation of the oil, fractionation at relatively lower crystallization temperature showed a considerable effect on fatty acid composition as well as triacylglycerol and diacylglycerol composition of liquid fractions compared to higher crystallization temperature. The olein and stearin fractionated at 4 degrees C had the best cold stability at 0 degrees C and sharper melting profile, respectively. PMID- 18387091 TI - Production of n-3 polyunsaturated fatty acid concentrate from sardine oil by immobilized Candida rugosa lipase. AB - This study was conducted to develop an immobilized-enzyme system to entrap lipase in chitosan-alginate-CaCl(2) beads for the purpose of concentrating n-3 polyunsaturated fatty acids (n-3 PUFAs) from sardine oil. Lipase was immobilized by an ionotropic gelatin method analyzed for characteristics. Optimum pH of immobilized lipase shifted from pH 7.0 to 6.0 and immobilized lipase showed higher stability against pH and temperature changes. Original sardine oil contained 38.1% n-3 PUFAs (25.2% 20:5n3 and 7.20% 22:6n3), and the concentration was significantly increased to 65.3% (40.2% 20:5n3 and 15.5% 22:6n3) with free lipase and to 64.8% (39.6% 20:5n3 and 15.3% 22:6n3) with immobilized lipase after 90 min of repeated hydrolysis. Fatty acid content of the free fatty acid (FFA) fraction of hydrolyzed oil showed that lipase preferably hydrolyzed 16:0, 16:1n7 and 18:0 accounting for 76.6% and 69.5% of total FFAs (after 1st and 2nd hydrolysis, respectively). This study shows that use of immobilized lipase systems for increasing n-3 PUFA concentration in sardine oil provides new processing opportunities for the industry. PMID- 18387092 TI - Pressurized liquid extraction of flavonoids from spinach. AB - Pressurized liquid extraction (PLE) using water and a 70:30 mixture of ethanol and water over the temperature range of 50 to 190 degrees C was used to extract flavonoids from dried spinach. The total phenolic content, antioxidant capacity, color, and browning indices of the extracts were also evaluated. PLE using a 70:30 mixture of ethanol and water was more effective than water in extracting flavonoids from spinach. Flavonoids were effectively extracted over the temperature band of 50 to 130 degrees C with water and 50 to 150 degrees C with ethanolic solvent. Levels of total phenolics and ORAC values increased with increasing extraction temperature, indicating that flavonoids were minor contributors to antioxidant capacity at elevated extraction temperatures. Browning of ethanolic extracts correlated highly with ORAC values over the temperature range of 50 to 190 degrees C, and the ORAC values of the large molecular weight fraction (> 1000 Da) increased linearly over the temperature range, indicating that Maillard polymers were the major contributors to antioxidant capacity. The results illustrate that PLE temperatures of < 130 degrees C for water or < 150 degrees C for ethanolic solvent may be used to extract flavonoids, followed by a high temperature (> 170 degrees C) extraction to generate antioxidant-rich moieties. PMID- 18387093 TI - Modification of granular corn starch with 4-alpha-glucanotransferase from Thermotoga maritima: effects on structural and physical properties. AB - Corn starch was converted using alpha-1,4-glucanotransferase from Thermotoga maritima (Tm alpha GT), a hyperthermophilic bacterium, without inducing gelatinization, and the structural changes and physical properties of the modified starches were investigated. Enzyme modification was induced at 65 degrees C for 8, 16, or 24 h, and the morphology of the modified starches was observed with light and scanning electron microscopy. Granule integrity was mostly maintained after enzyme treatment, although some granules were partially fragmented as evidenced by enlarged surface pores and some cracks. The modified starches had lower apparent amylose levels than raw starch. The molecular weights of amylose and amylopectin molecules in the treated starches were lower than those of raw starch, and the amount of branched molecules, which had much lower molecular weights, also increased in the treated starches. The chain-length distribution of amylopectin showed an increased number of shorter branched chains. The modified starches showed a wider melting temperature range and a lower melting enthalpy than that of raw starch. The X-ray diffraction pattern of the modified starches showed typical A-type starch peaks, but the relative crystallinities were lower than that of raw starch. The solubility and paste clarity of the modified starches were much higher than those of raw starch. The modified starch gels maintained their rigidity over the whole frequency range tested and showed thermoreversibility between 4 and 75 degrees C. These results suggest that Tm alpha GT can be used to produce granular corn starch, which contains amylose and amylopectin having lower molecular weights and a thermoreversible gelation property. PMID- 18387094 TI - Chemical and physical stability of protein and gum arabic-stabilized oil-in-water emulsions containing limonene. AB - An important flavor component of citrus oils is limonene. Since limonene is lipid soluble, it is often added to foods as an oil-in-water emulsion. However, limonene-containing oil-in-water emulsions are susceptible to both physical instability and oxidative degradation, leading to loss of aroma and formation of off-flavors. Proteins have been found to produce both oxidatively and physically stable emulsions containing triacylglycerols. The objective of this research was to determine if whey protein isolate (WPI) could protect limonene in oil-in-water emulsion droplets more effectively than gum arabic (GA). Limonene degradation and formation of the limonene oxidation products, limonene oxide and carvone, were less in the WPI- than GA-stabilized emulsions at both pHs 3.0 and 7.0. These data suggest that WPI was able to inhibit the oxidative deterioration of limonene in oil-in-water emulsions. The ability of WPI to decrease oxidative reactions could be due to the formation of a cationic emulsion droplet interface at pH 3.0, which can repel prooxidative metals, and/or the ability of amino acids in WPI to scavenge free radical and chelate prooxidative metals. PMID- 18387095 TI - Physicochemical properties of partially oxidized corn starch from bromide-free TEMPO-mediated reaction. AB - This study was conducted to determine the optimal temperature and time for the regiospecific oxidation of primary alcohol groups in corn starch with 2,2,6,6 tetramethyl-1-piperidinyl oxoammonium ion (TEMPO) and sodium hyphochlorite (NaOCl). The study also elucidated the molecular structure of fully oxidized corn starch (FOCS) prepared at optimum temperature and physicochemical properties of the partially (10%, 20%, and 30%) oxidized corn starches (POCS). The reaction time rapidly decreased up to 30 degrees C, and then gradually decreased. Selectivity, yield, and viscosity were drastically reduced at temperatures higher than 35 to 40 degrees C. Optimal oxidation temperature for the production of FOCS was determined as 35 degrees C. Regiospecific oxidation of the primary alcohol group without oxidation of the secondary alcohol group was confirmed in (13)C-NMR and IR spectra. Water-binding capacity, swelling power, solubility power, and transmittance of POCS increased as the degree of oxidation increased. Hardness, gumminess, and chewiness of corn starch gel containing POCS were not significantly different from those of native corn starch gel at 1-d storage, but the values of the starch gel containing POCS were smaller than those of the native starch gel after 1-d storage. However, springiness and cohesiveness did not differ significantly among the samples regardless of storage time. PMID- 18387096 TI - Discoloration and lipid deterioration of farmed giant catfish (Pangasianodon gigas) muscle during refrigerated storage. AB - Discoloration and lipid deterioration of farmed giant catfish (Pangasianodon gigas) muscle during 14 d refrigerated storage were investigated. Lipid deterioration, lipolysis, and lipid oxidation in both dorsal and ventral muscles increased as storage time increased. A progressive formation of primary lipid oxidation products monitored by the increase in conjugated dienes (CD) was observed (P < 0.05) and the increase in thiobarbituric reactive substances (TBARS), an index of secondary lipid oxidation products, was noticeable throughout the storage (P < 0.05). The pH of both dorsal and ventral muscles tended to increase as storage time continued (P < 0.05). A gradual increase in free fatty acid (FFA) formation was found within the first 10 d of refrigerated storage (P < 0.05), suggesting hydrolysis induced by lipases and phospholipases. However, a sharp decrease in FFA content was observed at the end of storage. Refrigerated storage also resulted in changes in redness index of both dorsal and ventral muscles. These changes were coincidental with the changes in metmyoglobin content. Therefore, the discoloration and lipid changes in giant catfish muscle during refrigerated storage depended on the muscle type and might be related to the difference in composition between dorsal and ventral muscles. PMID- 18387097 TI - Effect of heat on the distribution of fluorescently labeled plasminogen and plasminogen activators in bovine skim milk. AB - Differentially fluorescently labeled bovine plasminogen (PG-594) and human tissue and urokinase-type plasminogen activators (tPA-647 and uPA-546) were added to bovine skim milk to track the effect of heat on the location and concentration of these plasmin system components following acid precipitation or ultracentrifugation. In unheated milk, the majority (71.7% to 89.0%) of the added PG and PAs associated with casein micelles or acid curd, and PG-594 in the serum fraction was partially due to associations with nonsedimentable caseins. Heat treatment (85 degrees C for 16 s) significantly (P < 0.05) affected distribution of PG-594, tPA-647, and uPA-546, resulting in reduced concentrations of PG and PAs in the serum fractions and reciprocal increases in their levels in the nonsedimentable casein fractions. Overall, almost all of the added PG and PAs (95.9% to 97.5%) became associated with caseins following heat treatment. This is the 1st study to successfully use fluorescent labeling to quantify effects of heat on the location of plasmin components in skim milk. PMID- 18387098 TI - Biochemical and conformational changes of myosin purified from Pacific sardine at various pHs. AB - Biochemical and conformational changes of purified sardine myosin were investigated at various pHs. The purity of myosin, as determined by SDS-PAGE, was approximately 94.6%. One major band at 205 kDa, corresponding to myosin heavy chain, and 3 light chains at 31, 24, and 23 kDa were observed on the SDS-PAGE gel. The greatest myosin protein solubility was observed at pH 7 and remained constant up to pH 11. Sardine myosin showed no solubility at pHs 2.5 to 5.0. Three endothermic peaks were obtained for samples prepared at pHs 7 and 10, while no peaks were shown for pH 2 samples, indicating chemical denaturation of myosin occurred before thermal treatment. The greatest Ca(2+)-ATPase activity was observed at pH 7, while no activity was observed between pHs 2 and 5 and at pH 11. Total sulfhydryl content was not measured at pHs 2.5 to 4 while the greatest measure was obtained for samples at pH 5.5. Surface hydrophobicity was not detected from pHs 2.5 to 5.0; thereafter the content remained consistent through pH 11. Storage modulus, indicating the elastic element of myosin gels, was minimally affected at pH 2, indicating myosin was chemically denatured before the temperature sweep treatment. However, at pH 10, the thermal exposure of myosin, as evidenced by dynamic thermograms with deeper valleys at 40 to 60 degrees C, was noted, indicating myosin was not damaged by adjustment to pH 10 and therefore was still able to undergo thermal gelation. PMID- 18387099 TI - Measurement of the oxidation-reduction potential of cheddar cheese. AB - The objective of this study was to develop a method to measure the oxidation reduction (redox) potential of hard cheeses such as cheddar and to investigate the impact on this parameter of measurement temperature, and factors associated with electrochemical cell design such as distance between reference and working electrodes and depth into the cheese of the platinum electrodes. For this purpose, a novel, self-sealing, platinum working electrode was constructed which was thin and flexible enough to be inserted directly into the cheese sample. A calomel electrode was used as the reference electrode and the circuit was completed with a 3 M KCl salt bridge. The physical orientation of electrodes, such as distance between reference electrode and working electrode, had a substantial effect on equilibrium time for redox potential measurement. The time required for redox potential to reach equilibrium was 2 d in cheddar cheese and the optimum distance between the platinum and calomel electrodes was 2.5 cm. The fastest equilibration time was obtained when the working electrode was inserted 5 or 6 cm into the cheese. Temperature also had an important effect on redox potential. The shortest time to reach equilibrium of potential was at room temperature (20 degrees C), but it was not practical to keep cheese at this temperature for a period of 2 d. Therefore, redox measurement at 12 degrees C was recommended in spite of the longer equilibration time compared with room temperature. The results of this study suggest that the novel platinum working electrode allows reproducible measurement of the oxidation-reduction potential of cheddar cheese. PMID- 18387100 TI - Nitrogen-to-protein conversion factors for some cereal products in Japan. AB - To evaluate a practical method of determining more accurately conversion factors for calculating the protein contents of foods from the total nitrogen content, 19 cereal products found in Japan were analyzed for total nitrogen, amino acid nitrogen, and amide nitrogen, and then the nitrogen-to-protein conversion factors were calculated. The average conversion factors were 5.75 for rice, 5.81 for wheat, and 5.95 for others. These values, corresponding to the proportion of the amino acid residue to amino acid nitrogen recovered from 20 amino acids, were lower than the currently applied factors to these foods, except for wheat flour and amaranth. The use of this factor for estimating the protein content results in a considerable difference from the estimate based on amino acid residue concentrations, due to the wide variations in amino acid composition and to the presence of a significant level of nonprotein nitrogen. The distribution of the protein nitrogen recovered from the amino acids to total nitrogen averaged 93%. Adjusted conversion factors corresponding to the proportion of the amino acid residue to total nitrogen averaged 5.26 for rice, 5.47 for wheat, and 5.54 for other cereal products. Protein contents estimated using these factors are in good agreement with the contents defined as amino acid residues. PMID- 18387101 TI - Comparative structural, emulsifying, and biological properties of 2 major canola proteins, cruciferin and napin. AB - Canola is an economically important farm-gate crop in Canada. To further explore the potential of canola protein as value-added food and nutraceutical ingredients, a better understanding of fundamental properties of 2 major canola proteins is necessary. Two major protein components, cruciferin and napin, were isolated from defatted canola meal by Sephacryl S-300 gel filtration chromatography. SDS-PAGE showed that cruciferin consists of more than 10 polypeptides, and noncovalent links are more important than disulphide bonds in stabilizing the structural conformation. Napin consists of 2 polypeptides and is stabilized primarily by disulphide bonds. Purified cruciferin showed 1 major endothermic peak at 91 degrees C compared with that of 110 degrees C for napin. Emulsion prepared by cruciferin showed significant higher specific surface area and lower particle size than that of napin. The study indicated that the presence of napin could detrimentally affect the emulsion stability of canola protein isolates. Hydrolysates from cruciferin and napin showed potent angiotensin I converting enzyme inhibitory activity (IC(50): 0.035 and 0.029 mg/mL, respectively), but weaker than that of canola protein isolate hydrolysate (IC(50): 0.015 mg/mL). PMID- 18387102 TI - Mechanical properties of a Gelidium corneum edible film containing catechin and its application in sausages. AB - We prepared an edible Gelidium corneum (GC) film containing catechin and examined the microbial growth and quality change during storage of sausages packaged with the film. Incorporation of catechin in the film improved film tensile strength and water vapor permeability. The film's antimicrobial activity against Eschericha coli O157:H7 increased with increasing catechin concentrations and resulted in a decrease in the populations of the bacteria by 1.93 log CFU/g at 150 mg of catechin. For the sausage samples inoculated with E. coli O157:H7 and Listeria monocytogenes, the samples packed with the GC film showed a decrease in populations of E. coli O157:H7 and L. monocytogenes by 1.81 and 1.44 log CFU/g, respectively, compared to the control after 5 d of storage. In addition, the sausage samples packed with the GC film had lower degrees of lipid oxidation. The results suggest that sausages can be packed with GC film to extend shelf life. PMID- 18387103 TI - Carbon-centered radicals in isolated soy proteins. AB - Solid-state electron paramagnetic resonance (EPR) spectroscopy of commercial samples of isolated soy proteins (ISP) revealed a symmetrical free-radical signal typical of carbon-centered radicals (g= 2.005) ranging from 2.96 x 10(14) to 6.42 x 10(14) spins/g. The level of free radicals in ISP was 14 times greater than similar radicals in sodium caseinate, 29 times greater than egg albumin, and about 100 times greater levels than casein. Nine soy protein powdered drink mixes contained similar types of free radicals up to 4.10 x 10(15) spins/g of drink mix, or up to 6.4 times greater than the highest free-radical content found in commercial ISP. ISP samples prepared in the laboratory contained trapped radicals similar to the levels in commercial ISP samples. When ISP was hydrated in 2.3 mM sodium erythorbate or 8.3 mM L-cysteine, frozen and dried, the level of trapped free radicals increased by about 17- and 19-fold, respectively. The ESR spectrum of defatted soybean flakes contained overlapping signals from the primary free radical peak (g= 2.005) and a sextet pattern typical of manganese-II. The manganese signal was reduced in the laboratory ISP and very weak in the commercial ISP. PMID- 18387104 TI - Development and experimental validation of a frying model to estimate acrylamide levels in French fries. AB - In this study, a numerical model was developed to simulate frying of potato strips and estimate acrylamide levels in French fries. Heat and mass transfer parameters determined during frying of potato strips and the formation and degradation kinetic parameters of acrylamide obtained with a sugar-asparagine model system were incorporated within the model. The effect of reducing sugar content (0.3 to 2.15 g/100 g dry matter), strip thickness (8.5 x 8.5 mm and 10 x 10 mm), and frying time (3, 4, 5, and 6 min) and temperature (150, 170, and 190 degrees C) on resultant acrylamide level in French fries was investigated both numerically and experimentally. The model appeared to closely estimate the acrylamide contents, and thereby may potentially save considerable time, money, and effort during the stages of process design and optimization. PMID- 18387105 TI - Formulation of a cosurfactant-free O/W microemulsion using nonionic surfactant mixtures. AB - A cosurfactant-free O/W microemulsion composed of oil, a mixture of hydrophilic and hydrophobic surfactants, and water has been developed using food-grade components as a nutrient delivery system. We started our investigation to monitor the phase behavior of this system based on a hydrophilic surfactant (Tween 80). From a phase diagram, the weight ratio of 5.4:33.8:60.8 = oil:surfactant:water was selected as a combination ratio for the O/W microemulsion system. We also investigated the combination effect of different hydrophobic surfactants to Tween 80 on microemulsion formation. Use of hydrophobic surfactants with Tween 80 produced smaller droplets than Tween 80 alone. Rheological studies showed that all microemulsions followed shear-thinning behavior. The turbidity of microemulsions did not change after accelerated stability tests, indicating that this microemulsion system was stable under the given harsh conditions. When docosahexaenoic acid (DHA) oil was applied to this microemulsion system, the particle size and the turbidity were not significantly changed. Dilution with a different aqueous medium, either water or acidic fluid, did not significantly change the microemulsion turbidity. DHA oil incorporated in microemulsion showed higher oxidation stability than free DHA oil. PMID- 18387106 TI - A different perspective to study the effect of freeze, air, and osmotic drying on oil absorption during potato frying. AB - The objective of this article is to assess the effect of different dehydration pretreatments on oil absorption and illustrate how the different ways results can be reported may even drive to opposite conclusions. To do so, potato cylinders were blanched in hot water and dried until a moisture content of 62% (w.b.) by either freeze-drying, air drying, and osmotic drying with a sucrose solution or osmotic drying with a NaCl solution. Control (blanched) and dried potatoes were deep-fat fried at 170 degrees C for time periods between 1 and 5 min. Water removal and total oil uptake were determined and 2 oil fractions were distinguished: superficial oil and penetrated oil. Compared to the control, freeze dried samples increased oil uptake in 15.4% (d.b.) whereas air-dried samples reduced it in 11.2% (d.b.). Similarly, osmotic dehydrated samples showed a high reduction in oil uptake compared to the control (up to 27%[d.b.] when using a sucrose solution). However, this high decrease in oil absorption was attributed to the increase in solids content occurring during the osmotic dehydration process rather than a reduction in the amount of oil taken up. In fact, when the amount of oil absorbed per cylinder was determined, it was verified that oil uptake of osmotically dehydrated samples was even higher than the control, as opposed to what has been previously reported in the literature. These results highlight the importance of selecting an adequate basis to carry out comparisons properly. PMID- 18387107 TI - Cortical networks grown on microelectrode arrays as a biosensor for botulinum toxin. AB - Botulinum toxin (BoNT) is a potent neurotoxin produced by toxigenic strains of Clostridium botulinum. Botulinum toxin poses a major threat since it could be employed in a deliberate attack on the U.S. food supply. Furthermore, BoNT may be liberated in any insufficiently processed food containing a reduced oxygen atmosphere. Hence, rapid and reliable detection of BoNT in foods is necessary to reduce risks posed through food contamination. We present a BoNT biosensor employing living neural cultures grown in vitro on microelectrode arrays (MEAs). An MEA is a culture dish with a grid of electrodes embedded in its surface, enabling extracellular recording of action potentials of neural cultures grown over the array. Pharmaceutical grade BoNT A was applied to the media bath of mature cortical networks cultured on MEAs. Both spontaneous and evoked activities were monitored over 1 wk to quantify changes in the neural population produced by BoNT A. Introduction of BoNT A resulted in an increased duration and number of spikes in spontaneous and evoked bursts relative to control cultures. Increases were significant within 48 h of BoNT A dosage (P < 0.05). Application of BoNT A also induced unique oscillatory behavior within each burst that is reminiscent of early developmental activity patterns rather than the mature cultures used here. Three or more activity peaks were observed in 50% of the BoNT dosed cultures. Control cultures exhibited only a single activity peak. Thus activity of these cortical networks measured with MEAs could provide a valuable substrate for BoNT detection. PMID- 18387108 TI - Preparation of low-fat uptake doughnut by dry particle coating technique. AB - Soybean hull-wheat flour composites were prepared and used a doughnut formulation in order to decrease fat uptake during deep-fat frying. Soybean hulls were microparticulated by jet milling, and wheat flour was air-classified into coarse and fine fraction by an air classifying system. The coarse fraction of wheat flour was dry-coated with the microparticulated soybean hulls by a hybridization system. To investigate the effect on fat uptake, 4 different types of composites were prepared, which contained 0%, 1%, 5%, and 10% of soybean hulls. Doughnuts containing 1%, 5%, and 10% of microparticulated hulls decreased fat contents of 11.5%, 13.6%, and 35.8%, respectively. As soybean hulls content increased, hardness and crispiness increased. However, sensory evaluations demonstrated that there were no significant differences in appearance, flavor, crispiness, taste, and general liking (P < 0.05). Inner crust structures of doughnut showed slight reductions in cell size and improved cellular integrity with shrinkage in the cell membrane as the content of soybean hulls increased. These results show that microparticulated soybean hulls may form a protective layer during frying process, and this process could be used by the food industry for preparing doughnuts with reduced fat uptake. PMID- 18387109 TI - Effect of polymer surface modification on polymer-protein interaction via hydrophilic polymer grafting. AB - Surface modification of flat sheet ultrafiltration membranes, polyethersulfone (PES), was investigated to improve the hydrophilicity of the membrane surface thereby reducing adsorption of the proteins onto the membrane. Grafting of hydrophilic polymers onto UV/ozone-treated PES was used to improve the hydrophilicity of the commercial PES membranes. Hydrophilic polymers, that is, poly(vinyl alcohol) (PVA), polyethylene glycol (PEG), and chitosan, were employed to graft onto PES membrane surfaces because of their excellent hydrophilic property. The surfaces of modified PES membranes were characterized by contact angle measurement, FTIR, and AFM. The FTIR spectra indicated that PES membranes were successfully modified by grafting of the hydrophilic polymers. The modified PES membranes showed 20% to 50% reduction in contact angle measurements in comparison with those of the virgin PES membrane. The tapping mode AFM technique was employed to investigate the changes of surface topography, cross-section, and root mean square roughness of the modified PES membrane surfaces. The modified PES membranes showed elevated roughness (ranging from 7.0 to 25.7 nm) compared with that of the virgin PES membrane (2.1 nm). It is concluded that grafting of PVA, PEG, or chitosan onto UV/ozone-treated PES membranes increases hydrophilicity and lowers protein adsorption by 20% to 60% compared to the virgin PES membrane. Among the 3 hydrophilic polymers studied, PEG showed the most favorable result in terms of contact angle and protein adsorption. PMID- 18387110 TI - A soybean cultivar lacking lipoxygenase 2 and 3 has similar calcium bioavailability to a commercial variety despite higher calcium absorption inhibitors. AB - The aim of this study was to evaluate calcium bioavailability of a new soybean variety without 2 lipoxygenases with better taste and flavor than a commercial variety containing all 3 isozymes. Using the femur (45)Ca uptake method, calcium absorption from a new Brazilian variety, UFV-116, was compared to a common Brazilian variety, OCEPAR 19. Male Sprague-Dawley growing rats weighing 150 to 170 g (10/group) received test meals of whole fat soy flour prepared from UFV-116 or OCEPAR-19 seeds labeled with 10 microCi of (45)Ca. Femurs were removed after 48 h for determination of (45)Ca uptake. Calcium fractional absorption was equivalent between the 2 varieties. The higher oxalate:calcium molar ratio and the higher content of oxalate and phytate (P < 0.05) found in the UFV-116 variety did not affect calcium absorption. Therefore, the new variety is a comparable source of high bioavailable calcium. PMID- 18387111 TI - The degree of polymerization of inulin-like fructans affects cecal mucin and immunoglobulin A in rats. AB - Cecal amounts of mucin and immunoglobulin A (IgA) were examined through the cecal fermentation pattern in Wistar (WS) or Sprague-Dawley (SD) rats fed inulin-type fructans differing in their degree of polymerization (DP). The animals were fed a control diet or a diet containing one of the fructans with an average DP of 4, 8, 16, or 23, at 60 g/kg diet for 10 d. Cecal fermentation products substantially differed between WS and SD rats fed DP8 fructan, with short-chain fatty acids (SCFAs) as the major organic acids in the former but lactate predominating in the latter. Cecal fermentability of fructans in both strains generally decreased with increasing DP of fructans, and this was especially manifest in reduction of the amounts of lactate in DP16 and 23. In WS rats, cecal mucin and IgA were greater in all fructan groups than in the control group. In SD rats, cecal mucin was greater only in the DP8, 16, and 23 groups as compared to the control group, while IgA was greater in the DP4 and 8 groups. In both strains, cecal mucin correlated with the sum of cecal SCFAs, but not with lactate, succinate, or total organic acids. In contrast, only cecal lactate correlated with cecal IgA in both strains. The present study shows that the different fermentation patterns of fructans affect cecal mucin and IgA; mucin is likely to respond to cecal SCFA production, whereas IgA increases when fermentation occurs rapidly and lactate is a major fermentation product. PMID- 18387112 TI - Relationships between ultimate pH and microbial, chemical, and physical characteristics of vacuum-packaged pork loins. AB - This study evaluated the effect of ultimate pH (pHu) of pork on shelf life based upon microbial growth, drip loss, and oxidative rancidity (2-thiobarbituric acid [TBA] procedure) in vacuum-packaged loins stored at 4 degrees C. Glucose and lactate concentrations of the pork loins were also measured. Thirty-six pork loins (pH = 5.56 to 6.57) were collected at a commercial slaughter facility 1-d postslaughter. All pigs were from the same genetic line. Loins were grouped by pH (group: pH range): A: 5.55 to 5.70, B: 5.71 to 5.85, C: 5.86 to 6.00, D: 6.01 to 6.15, and E: > 6.16. They were analyzed at days 0, 6, 14, 24, and 34. For aerobic plate counts, groups A and B were significantly lower than C through E, while psychrotrophic or Enterobacteriaceae counts of groups A and A through C were significantly lower than groups B through E and D and E, respectively. Lactic acid bacteria counts were not significantly influenced by pHu. Group A had higher glucose concentrations than groups C through E and higher lactate concentrations than groups D through E on most sampling days. Group A had a higher TBA value than group E at days 0 and 34. Group A displayed greater drip loss than groups D and E at day 6 and groups B through D on days 24 and 34. Based on the microbial and drip loss results, a pork loin pHu of 5.8 to 5.9 appears to be optimum to provide a vacuum-packaged shelf life of at least 24 d with minimum drip loss. PMID- 18387113 TI - Influence of probiotic Lactobacillus acidophilus and L. helveticus on proteolysis, organic acid profiles, and ACE-inhibitory activity of cheddar cheeses ripened at 4, 8, and 12 degrees C. AB - The influence of adjunct bacteria on composition of cheeses, organic acid profiles, proteolysis, and ACE-inhibitory activity during ripening at 4, 8, and 12 degrees C for 24 wk was investigated. cheddar cheeses were made with starter lactococci (control), Lactobacillus acidophilus L10, and starter lactococci (L10), and L. acidophilus L10, L. helveticus H100, and starter lactococci (H100). The counts of L. acidophilus in L10 cheeses remained at >10(6) colony forming units (CFU)/g after 24 wk of ripening at 4, 8, and 12 degrees C. Concentrations of lactic, acetic, and propionic acids of the L10 and H100 cheeses were significantly higher than those of the control cheeses after 24 wk of ripening (P < 0.05). Proteolysis of the cheeses was improved as the ripening temperature increased. Water-soluble nitrogen, trichloroacetic acid soluble nitrogen, and phosphotungstic acid soluble nitrogen of L10 and H100 cheeses were significantly higher than those of the control cheeses (P < 0.05). Increase in ripening temperature from 4 degrees C to 8 and 12 degrees C increased the percentage of ACE inhibition. The IC(50) value among cheeses ripened at 4, 8, and 12 degrees C, however, was not significantly different (P > 0.05). Hence, probiotic L. acidophilus L10 can be added into cheddar cheeses to improve proteolysis and ACE inhibitory activity. PMID- 18387114 TI - Phytosterol effects on milk and yogurt microflora. AB - Phytosterols play a major role in functional foods. Their ability to reduce serum cholesterol in humans has been widely proven and they are now being added to various dairy based products. The present study investigated the potential antimicrobial activity of phytosterols in milk and their effect on yogurt starter cultures. A commercial phytosterol (0.26% to 1.8%, w/v) preparation (CPP) consisting of ss-sitosterol, campesterol, sitostanol, and campestanol had no effect on the standard plate count (SPC) and psychrotroph population in pasteurized milk stored at 4 degrees C. In addition, a challenge study employing Pseudomonas spp. in milk at 4 to 7 degrees C confirmed that the CPP was not antimicrobial. However, the addition of a dispersible CPP consisting of 0.72% phytosterol containing 0.02% to 0.03% sodium stearoyl lactylate (SSL) did appear to affect the SPC and psychrotrophic bacteria in refrigerated milk. The dispersible preparations did not, however, inhibit the growth of Pseudomonas. An investigation into the antimicrobial activity of SSL revealed that it alone had no effect on the SPC in milk. The CPP had no effect on growth and acid development by Lactobacillus bulgaricus and Streptococcus thermophilus during yogurt production at 33 degrees C and storage at 4 degrees C for 30 d. This is seen as a beneficial feature since growth and acid development by these organisms are crucial for yogurt quality. Saccaromyces cerevisiae and Aspergillus ochraceous added to yogurt as typical contaminants also were not inhibited. While the CCP was somewhat antimicrobial when formulated with dispersing agents, it otherwise had no antimicrobial activity. PMID- 18387115 TI - Biodegradable polylactic acid polymer with nisin for use in antimicrobial food packaging. AB - Biodegradable polylactic acid (PLA) polymer was evaluated for its application as a material for antimicrobial food packaging. PLA films were incorporated with nisin to for control of foodborne pathogens. Antimicrobial activity of PLA/nisin films against Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Enteritidis were evaluated in culture media and liquid foods (orange juice and liquid egg white). Scanned electron micrograph and confocal laser microscopy revealed that nisin particles were evenly distributed in PLA polymer matrix on the surface and inside of the PLA/nisin films. PLA/nisin significantly inhibited growth of L. monocytogenes in culture medium and liquid egg white. The greatest inhibition occurred at 24 h when the cell counts of L. monocytogenes in the PLA/nisin samples were 4.5 log CFU/mL less than the controls. PLA/nisin reduced the cell population of E. coli O157:H7 in orange juice from 7.5 to 3.5 log at 72 h whereas the control remained at about 6 log CFU/mL. PLA/nisin treatment resulted in a 2 log reduction of S. Enteritidis in liquid egg white at 24 degrees C. After 21 d at 4 degrees C the S. Enteritidis population from PLA/nisin treated liquid egg white (3.5 log CFU/mL) was significantly less than the control (6.8 log CFU/mL). E. coli O157:H7 in orange juice was more sensitive to PLA/nisin treatments than in culture medium. The results of this research demonstrated the retention of nisin activity when incorporated into the PLA polymer and its antimicrobial effectiveness against foodborne pathogens. The combination of a biopolymer and natural bacteriocin has potential for use in antimicrobial food packaging. PMID- 18387116 TI - Influence of autoinducer-2 (AI-2) and beef sample extracts on E. coli O157:H7 survival and gene expression of virulence genes yadK and hhA. AB - Bacterial cell-to-cell communication is mediated by autoinducer (AI) molecules such as AI-2 and has been reported to regulate gene expression in Escherichia coli O157:H7. We have previously shown that ground beef contains compounds that can inhibit sensing of AI-2 like activity. The hypothesis of this study was that AI-2 activity observed in conditioned medium (CM) will enhance E. coli O157:H7 survival and expression of virulence genes, whereas compounds inhibitory (such as those present in ground beef extracts) to AI-2 activity will negate these effects. E. coli O157:H7 luxS mutant strain VS 94 (incapable of synthesizing AI 2) was employed in these studies. The survival of this enteric bacterial pathogen as a function of AI-2 activity and the presence of AI-2 inhibitory compounds was studied at 4 degrees C. The number of survivors in the presence of AI-2 was significantly higher compared to the absence of AI-2, and the addition of ground beef extracts to conditioned medium negated the influence of AI-2 activity. Autoinducer AI-2 upregulated selected genes virulence genes (yadK, and hha), whereas the ground beef extract reversed the effect of AI-2 on the expression of the selected genes. PMID- 18387117 TI - Autoinducer-2 activity of gram-negative foodborne pathogenic bacteria and its influence on biofilm formation. AB - This study evaluated whether autoinducer-2 (AI-2) activity would be associated with biofilm formation by Salmonella and Escherichia coli O157:H7 strains on food contact surfaces. In study I, a Salmonella Typhimurium DT104 strain and an E. coli O157:H7 strain, both AI-2 positive, were individually inoculated into 50 mL of Luria-Bertani (LB) or LB + 0.5% glucose (LBG) broth, without or with stainless steel or polypropylene (Salmonella) coupons. At 0, 14 (Salmonella), 24, 48, and 72 h of storage (25 degrees C), cells in suspension and detached cells from the coupons, obtained by vortexing, were enumerated on tryptic soy agar. In study II, a Salmonella Thompson AI-2-positive strain and an AI-2-negative strain, and an E. coli O157:H7 AI-2-positive strain and an AI-2-negative strain were inoculated into LB broth with stainless steel coupons. Cells were enumerated as in study I. In both studies, AI-2 activity was determined in cell-free supernatants. Cell numbers of S. Typhimurium DT104 on biofilms were higher (P < 0.05) in LB than those in LBG, while the E. coli O157:H7 strain showed no difference (P>or= 0.05) in biofilm cell counts between LB and LBG after storage for 72 h. Both S. Typhimurium DT104 and E. coli O157:H7 strains produced higher (P < 0.05) AI-2 activity in LBG than LB cell suspensions. Cell counts of AI-2-positive and negative S. Thompson and E. coli O157:H7 strains were not different (P>or= 0.05) within suspensions or coupons (study II). The results indicated that, under the conditions of this study, AI-2 activity of the pathogen strains tested may not have a major influence on biofilm formation on food contact surfaces, which was similar between AI-2-positive and -negative strains. PMID- 18387118 TI - Fresh-cut lettuce in modified atmosphere packages stored at improper temperatures supports enterohemorrhagic E. coli isolates to survive gastric acid challenge. AB - Incidences of foodborne outbreaks involving enterohemorrhagic Escherichia coli strains with mutations in a key regulatory gene, rpoS, have been reported. Incentives, if any, for losing this regulatory function are not clear since the RpoS regulator is required for the expression of several environmental stress tolerance genes. RpoS also positively regulates 2 of the 3 acid-resistance systems of E. coli under aerobic growth conditions and enables the pathogen to survive gastric acid challenge. We selected 7 enterohemorrhagic E. coli isolates, 6 of which are known to carry defective rpoS gene, and then analyzed resistance to synthetic gastric juice after the strains were inoculated on fresh-cut lettuce and stored under modified atmosphere packaging (MAP) conditions. Subatmospheric oxygen partial pressures in MAP enabled all 6 rpoS-defective isolates to induce acid resistance over the 8-d storage period if the temperature was >or= 15 degrees C. No acid resistance was induced for MAP-stored lettuce left at temperatures or=1 month, 463 (85%; 95% CI 79-91; design effect 3.6) were classified as vaccinated. Amongst the study dogs, dogs aged 1-11 months were the higher risk of dogs not being vaccinated (OR = 8.2; 95% CI 4.3-15.6; P < 0.01). Almost two-thirds of the study dogs were allowed to roam freely throughout the day or in part. Community education efforts should address the importance of dog ownership and movement restriction, and the need to vaccinate young dogs. PMID- 18387139 TI - Chlamydophila psittaci infections in humans during an outbreak of psittacosis from poultry in Germany. AB - In 2005, an outbreak of severe respiratory disease in a mixed poultry flock that was infected with Chlamydophila (C.) psittaci led to dissemination of the infection to at least 100 small poultry farms in 11 districts of Central Germany. At the same time, a total of 24 persons in contact with poultry from one of the flocks reported flu-like symptoms to their physician, thus suggesting zoonotic transmission. Within 3 weeks, seven individuals had to be hospitalized, with three of them requiring intensive care. Analysis of ompA sequences from chlamydial isolates and directly from clinical samples revealed the presence of both genotype A and E/B of C. psittaci at the source of the outbreak and in contact flocks. Genotype A was also detected in the three severely ill patients. The findings of the present study demonstrate the high zoonotic potential of avian chlamydiae. To ensure speedy eradication of psittacosis in poultry flocks and effective treatment of infected humans, fast, sensitive and species-specific detection of the causative agent is essential, as well as close collaboration between regional public health services, attending physicians and the diagnostic laboratories involved. PMID- 18387140 TI - Human seroprevalence to Coxiella burnetii (Q fever) in Northern Ireland. AB - Despite the widespread prevalence of infection with Coxiella burnetii, there have been few large population-based studies examining the epidemiology of this infection. The aim of this study was to examine the distribution and determinants of C. burnetii past infection in Northern Ireland (NI). Coxiella burnetii phase II specific IgG antibodies were measured by enzyme-linked immunosorbent assay in stored serum from 2,394 randomly selected subjects, aged 12-64, who had participated in population-based surveys of cardiovascular risk factors performed in 1986 and 1987. The overall prevalence of C. burnetii antibody positivity was 12.8%. The prevalence of sero-positivity was slightly higher in males than that in females (14.3% versus 11.2%, P = 0.02). Sero-positivity was low in children (<10%), increasing to 19.5% and 16.4% in males and females, respectively, in the 25-34 age group and subsequently remaining fairly steady with increasing age. Sero-positivity among farmers, at 48.8%, was significantly higher than the general population. More sero-positive than sero-negative women had a history of a miscarriage or still-birth (19.5% versus 9.8%, P < 0.001). In conclusion, this study demonstrated a high prevalence of evidence of past C. burnetii infection in NI. Associations between past C. burnetii infection and age, sex, social class, occupation and reproductive history were seen. We estimate that 20% of Q fever infections in NI occur in farmers. PMID- 18387141 TI - Antimicrobial resistance of Salmonella enterica isolates from apparently healthy and clinically ill finishing pigs in Spain. AB - This study was the first conducted in Spain to evaluate the occurrence of antimicrobial resistance and multi-resistance in Salmonella isolates recovered from finishing pigs from Spanish swine farms distributed over the whole country. For this purpose, 290 Salmonella isolates recovered from apparently healthy finishing pigs in a farm-based cross-sectional study and 192 Salmonella isolates recovered from faecal samples of finishing pigs suffering from diarrhoea were investigated. Resistance to a panel of 17 antimicrobials was determined using a broth microdilution technique. Resistance was a common finding and was detected in 90.3% of the Salmonella isolates from apparently healthy finishing pigs and 95.3% of the Salmonella isolates from clinically diseased finishing pigs. Resistance was particularly high among isolates of serogroup B and serovars Typhimurium and its monophasic variant S. 4,5,12:i:-. Higher frequencies of resistance were found to tetracycline, sulphamethoxazole, streptomycin, spectinomycin, ampicillin, chloramphenicol and trimethoprim-sulphamethoxazole. Less than 10% of the isolates were resistant to amoxicillin/clavulanic acid, neomycin, cephalotin, apramycin and gentamicin. Resistance to ciprofloxacin, colistin and ceftiofur was rare (under 1%). Multi-resistance, defined as resistance to four or more drugs, was detected in more than 50% of the isolates. Although multi-resistance was particularly frequent among isolates of S. Typhimurium, it was also high among other serovars as Bredeney and the S. Typhimurium monophasic variant. 4,5,12:i:-. PMID- 18387142 TI - Prevalence of bacterial pathogens and their anti-microbial resistance in Tilapia and their pond water in Trinidad. AB - In Trinidad, Tilapia (Oreonchromis spp.) is one of the most important fresh water food fish and the number of farms has been increasing annually. A study was conducted in the local tilapia industry to determine the microbial quality of pond water, prevalence of bacterial pathogens and their anti-microbial resistance using the disk diffusion method. Seventy-five apparently healthy fish and 15 pond water samples from three of the four commercial tilapia fish farms in the country were processed. The 202 bacterial isolates recovered from fish slurry and 88 from water, belonged to 13 and 16 genera respectively. The predominant bacteria from fish slurry were Pseudomonas spp. (60.0%), Aeromonas spp. (44.0%), Plesiomonas (41.3%) and Chromobacterium (36.0%) (P < 0.05; chi(2)) compared with isolates from pond water where Bacillus spp. (80.0%), Staphylococcus spp., Alcaligenes spp. and Aeromonas spp. (60.0%) were most prevalent (P < 0.05; chi(2)). Using eight anti-microbial agents, to test bacteria from five genera (Aeromonas, Chromobacterium, Enterobacter, Plesiomonas and Pseudomonas), 168 (97.1%) of 173 bacterial isolates from fish slurry exhibited resistance to one or more anti microbial agents compared with 47 (90.4%) of 52 from water (P > 0.05; chi(2)). Resistance was high to ampicillin, 90.2% (158 of 173), erythromycin, 66.5% (115 of 173) and oxytetracycline, 52.6%, (91 of 173) but relatively low to chloramphenicol, 9.8% (17 of 173) and sulphamethoxazole/trimethoprim, 6.4% (11 of 173) (P < 0.05; chi(2)). For pond water isolates, the frequency of resistance across bacterial genera ranged from 75% (nine of 12) for Chromobacter spp. to 100% found amongst Enterobacter spp. (six of six), Plesiomonas spp. (nine of nine) and Pseudomonas spp. (eight of eight) (P < 0.05; chi(2)). Resistance was generally high to ampicillin, 78.8% (41 of 52), erythromycin, 51.9% (27 of 52) and oxytetracycline, 34.5% (18 of 52) but low to sulphamethoxazole/trimethoprim, 7.7% (four of 52) and norfloxacin, 3.8% (two of 52) (P < 0.05; chi(2)). It was concluded that the rather high prevalence of bacterial pathogens in tilapia along with their high prevalence of resistance to anti-microbial agents might pose therapeutic problems as well as health risk to consumers. The microbial presence and their anti-microbial resistance in the tilapia industry are being reported for the first time in the country. PMID- 18387143 TI - Distribution of virF/lcrF-positive Yersinia pseudotuberculosis serotype O:3 at farm level. AB - The distribution and persistence of pathogenic, virF/lcrF-positive Yersinia pseudotuberculosis were investigated in pigs and in the pig house environment during rearing to determine possible contamination routes of early infections. Based on Y. pseudotuberculosis-positive tonsils of slaughter pigs in our previous study, Y. pseudotuberculosis-positive animals were traced back to the farms. Eight farms were visited from 6-10 months later, and a total of 155 pooled and six individual faecal samples from pigs and 116 pooled environmental samples were collected for analysis by different culture methods. Four of the eight farms were found to be Y. pseudotuberculosis-positive. All positive faecal samples were obtained from fattening pigs, with prevalence varying from 5% to 71% on positive farms. Sows, boars and suckling piglets were Y. pseudotuberculosis-negative on all farms. Most Y. pseudotuberculosis-positive farms (three of four) were on a one-site production system, which had a higher prevalence of Y. pseudotuberculosis (5-26%) among fattening pigs than the all-in, all-out system (1-5%). All Y. pseudotuberculosis isolates belonged to serotype O:3 and carried the virF/lcrF gene on the virulence plasmid. Biotypes 2 and 3 were involved, the latter in one isolate and not being previously reported in pigs. Altogether 53 isolates from 16 positive samples were characterized with pulsed-field gel electrophoresis (PFGE). Using SpeI, NotI and XbaI enzymes, four, three and two different PFGE patterns were obtained respectively. A total of nine different genotypes were identified when the profiles of the enzymes were combined. The most common genotypes were gIV, found on three, and gXII, found on two of the four Y. pseudotuberculosis-positive farms. The same genotypes previously detected in pig tonsils were present in pig faeces from the same farm, indicating that some Y. pseudotuberculosis strains can persist in the pig house environment. PMID- 18387144 TI - Longitudinal study of Salmonella infection in Italian farrow-to-finish swine herds. AB - A longitudinal study of Salmonella enterica infection was carried out in five Italian farrow-to-finish swine herds previously known to be infected by Salmonella. Five litters were randomly selected from each herd and in each litter six piglets were randomly selected and individually identified. Thus, the study included 30 pigs from each farm. At weaning, individual blood samples were collected for serological examination from all selected piglets and on the same day from all sows in the farrowing unit. Piglets were bled again at approximately 60, 90, 150, 210 and 270 days of life whereas the last blood sample was collected at slaughtering. In one of the herds, in which the duration of productive cycle was about 12 months, the last blood samples were collected at 350 days of life. With the same time scheduling, five pen pooled faecal samples were collected from each herd for bacteriological examination. At slaughtering, mesenteric lymph nodes were collected from each ear-tagged pig. Sero-prevalence (cut off S/P ratio 0.25) in sows varied from 93.8% to 100%. In four herds, sero-prevalence in piglets showed a similar profile with complete decline of maternal antibodies at day 60 and clear sero-conversion between day 90 and day 150. In one herd, sero conversion was observed earlier and 56% of piglets were positive at day 90. The peak of sero-prevalence was observed between day 210 and day 270. Sero-prevalence at slaughtering varied from 66% to 100%. Salmonella was isolated from faecal samples in four of five herds. No Salmonella was isolated from mesenteric lymph nodes at slaughter in two of the herds. Culture prevalence from mesenteric lymph nodes in the other three herds ranged from 3.3% to 30%. This longitudinal study provides original information about epidemiological dynamics of Salmonella enterica infection in Italian swine herds in consideration of the unique extended fattening period typical of the Italian production. PMID- 18387145 TI - Suicide ideation among later elementary school-aged youth. AB - Suicide is extremely rare among persons under age 15 years old. Nationwide, it has been reported that 18% of students in grade 6 had thoughts of killing themselves. The Social Disintegration Model (SDM) summarizes intra-personal, interpersonal, peer network, physical and heath risk behaviour characteristics that may influence suicide ideation. As part of a larger study to test a family strengthening programme, 179 later elementary school children from two cities responded to 20 items about their participation in health risk behaviour. Sixteen youth indicated they had thought of killing themselves. Based on the SDM, these 16 youth were compared with the larger sample on measures of ways of coping; family communication, functioning and caring; school connectedness, pubertal development; and alcohol use and weapon carrying. Youth who responded positively to the 'thought of killing self' screening question, felt less connected to their school, used more internalizing behaviours and reported less cohesion, open communication, supervision and family caring than youth who answered no. Youth who thought of killing themselves may benefit from additional school support to feel more comfortable and connected at school. Community resources may help parents modify child-rearing behaviours. Building communication among parents and youth may prevent suicidal behaviour among young adolescents. PMID- 18387146 TI - Out-of-area placements in Scotland and people with learning disabilities: a preliminary population study. AB - People with learning disabilities have a different pattern of disease from the general population and high health needs that are frequently unidentified and unmet. Many require responses from general and specialist health services. A picture is emerging of some people with learning disabilities, often with complex care needs, moving from their home area on what is being termed, out-of-area placements, to receive specialist care. However, within the learning disability population, limited research has been undertaken and the impact on health services is unknown. Data were collected from health and social care providers to identify people with learning disabilities moving in and out of services across Scotland. Further data about the consequences and impact of out-of-area placements were gathered in one geographical area using focus group methodology. The results suggest that people with learning disabilities are moving in, out and across Scotland, often as a result of breakdown of local care arrangements or because of lack of specialist resources. Planning, service development and effective communication need to be in place to address the needs of this increasing and ageing population. PMID- 18387147 TI - Decision-making needs of patients with depression: a descriptive study. AB - The study's purpose was to explore the decision-making needs of patients considering treatment options for their depression. Semi-structured interviews were guided by the Ottawa Decision Support Framework. Of 94 participants, 67 were uncertain about their decision. Common decisions identified were whether or not to take medications, attend support groups, undergo electroconvulsive therapy, and location of care. Those feeling certain were more likely to have made a decision (RR 1.37; 95% CI: 1.05, 1.78). However, 40 patients who had 'made a decision' in the recent past were uncertain about their decision. Compared with those who were certain, the uncertain group felt less informed (2.65 vs. 1.64; P < 0.001), less supported (2.63 vs. 1.88; P < 0.001) and less clear about how they valued the benefits and risks of options (2.57 vs. 1.69; P < 0.001). Other influential factors included concerns about confidentiality, distress from depression, embarrassment, panic attacks and lack of energy. Few patients wanted to defer decision making to their physician (n = 8) or family (n = 1). To support decision making, participants identified the need for: discussions with their psychiatrist, nurse or family doctor; access to printed information; and information provided by health professionals and health societies. PMID- 18387148 TI - An investigation into the aspirations and experiences of newly appointed dual diagnosis workers. AB - This qualitative, exploratory study was designed to explore a sample of eight recently appointed dual diagnosis workers' (DDWs) perceptions of their new role and function in Northern Ireland (NI). A semi-structured interview was used and respondents were assured that their anonymity/rights would be protected. All of the narratives were shown to the respondents for their approval prior to going to press. The transcripts were analysed by using a tried and tested analytical framework. Seven key categories emerged from the findings relating to the DDWs perceptions of their: (1)understanding of the term dual diagnosis; (2) hopes; (3) fears; (4) support in their new role; (5) key clinical issues; (6) the positioning of the service; and (7) their overall role and function. This is a new and important area of work in NI. However, to date, no research has been carried out on the topic in the province. Consequently, the findings from this small study could go some way towards helping to shape the future direction of, and bring about some universality to the provision of the dual diagnosis service within different National Health Service Trusts in NI. Further research is required on this new and growing service as well as on the service users' perceptions of the care provided by DDWs. The study will be followed up on an annual basis for 3 years to provide longitudinal data. Generalization of findings requires caution because of the small sample size. PMID- 18387149 TI - Does the position of restraint of disturbed psychiatric patients have any association with staff and patient injuries? AB - The aim of this study was to examine the risk of injury among patients and staff following involvement in a restraint episode in relation to restraint position (standing, supine or prone) and other aspects of the pre-incident behaviour including perceived causation. Mixed effects logistic regression was used to estimate the relative odds of injury to staff or patient in a series of 680 restraint episodes involving 260 patients in an adult mental health service in England between 1999 and 2001. There was no statistically significant association between patient injury and restraint position in this sample, but a prone restraint position was weakly associated with staff injury. Staff injury was most likely when an actual assault had occurred prior to the incident. Patient injury was more likely when the patient had self-harmed, had been abusing substances and had used a weapon prior to the incident, and less likely when the patient was showing signs of frustration with their environment. The use of prone restraint may be weakly associated with an increased risk of injury to staff. However, other aspects of the incident are stronger predictors and should be considered when planning training for front line staff. PMID- 18387150 TI - Translating theory into practice: results of a 2-year trial for the LEAD programme. AB - This paper presents data for 2 years of a continuing study aimed to reduce the risk of first-time involvement by minority youth with the juvenile justice system. A quasi-experimental design was used to test a 14-week expressive art curriculum (LEAD: leadership, education, achievement and development) implemented in two rural communities. A total of 70 African American youth participated in the programme over a 2-year period. Pre- and post-test differences were examined for protective factors, behavioural self-control, self-esteem and resilience measures. When compared with the after-school programme (control group), youth in the LEAD programme in both communities had more dramatic increases in post-test scores following the intervention. The combined data from year 1 and year 2 provide positive findings in support of LEAD as a prevention programme for young offenders. The placement of the programme within an African American church in year 2 improved the processes of the LEAD programme and seemed to provide a better fit with the original design of the programme, highlighting the importance of the context, in which the programme was provided. PMID- 18387151 TI - Intimate partner violence as experienced by men. AB - The purpose of this qualitative, phenomenological study is to describe intimate partner violence as experienced by men and to formulate the common structure of meanings of experiences of men exposed to intimate partner violence. The data were gathered by open-ended interviews with 10 (n = 10) men. The participants had experiences in intimate partner violence. All the 10 men were interviewed twice and they were allowed to tell their experiences as they liked. The data were analysed by applying the method developed by Colaizzi. The experiences of men formulated three main themes describing the factors leading to intimate partner violence, being a victim of intimate partner violence and own violent behavior. The main themes are named as follows: (1) a burden on the pair relationship; (2) face to face with violence; and (3) from denying violence to striving towards the truth. The research results enhance the understanding of the experiences of men. The information may be useful in promoting women's, men's and families' health, developing work on violence and in the education of professionals in social and health care. The results may also help women to understand men's experiences in a pair relationship. PMID- 18387152 TI - Clinical supervision in cognitive behavioural psychotherapy: development of a model for mental health nursing through grounded theory. AB - This study focuses on the development of a cognitive behavioural model of supervision for mental health nurses. The study utilized a grounded theory approach with cognitive behavioural psychotherapy training course directors. The aim was to more fully understand cognitive behavioural supervision from the perspective of expert supervisors, and develop a model of supervision for mental health nurses who are also cognitive behavioural psychotherapists. For this purpose, 16 course directors were interviewed in-depth, with data analysis taking place after each interview. Through a process of inductive reasoning, core categories were identified from the participants themselves. The relationships between the categories are described. The findings are discussed in terms of a new model that can be used to underpin cognitive behavioural psychotherapy supervision in mental health nursing. PMID- 18387153 TI - Creating communities in cyberspace: pro-anorexia web sites and social capital. PMID- 18387154 TI - Time to wake up and smell the coffee: antipsychotic medication and schizophrenia. PMID- 18387157 TI - Therapeutic plasma exchange combined with immunomodulating agents in secondary progressive multiple sclerosis patients. AB - Multiple sclerosis (MS) is the most common cause of neurological disability. Therapeutic plasma exchange (TPE) has been used in the management of patients with MS with equivocal efficacy. With this work we would like to present our experience with 10 patients (seven male and three female, mean age 34 years [range 27-53 years]) with secondary progressive MS, who were treated with immunomodulating agents and who also underwent TPE. The patients' expanded disability status scale (EDSS) score of entry to the study varied from 5.0 to 6.5. One year before study entry all patients showed a marked deterioration (12 months before starting TPE they had been rated 4.5-5.5 on the EDSS). TPE was performed three times a week for two weeks and thereafter once a week, or once a month for the stable patients. The machine used was the Cobe Spectra and albumin 5% was the replacement fluid. Peripheral veins were used in nine patients and indwelling vascular access was required in one patient. Eighteen months later, patients stopped taking the immunomodulating agent therapy and continued only with TPE. No side-effects occurred during the TPE session. After 36 months of TPE therapy, five patients were stabilized in their disability, while two patients showed a minor progression of the disease (an additional 0.5 degree in disability as determined by the EDSS). No relapses occurred during TPE. Three patients stopped the therapy: one patient because of persistent nausea and two patients for reasons unrelated to TPE. Periodic TPE was associated with reduced accumulation of neurological deficits (as documented by EDSS) in patients with secondary progressive MS. PMID- 18387158 TI - Autoantigen estimation and simple screening assay against cardiodepressant autoantibodies in patients with dilated cardiomyopathy. AB - The objective of this study was to identify the cardiodepressant autoantibodies that could directly influence left ventricular ejection fraction (LVEF) in patients with dilated cardiomyopathy (DCM), as well as to establish a simple screening method for these antibodies. Not only acute hemodynamic but also chronic prognosis improvements were reported with immunoadsorption in some patients with DCM. Various antibodies determined by immunohistochemistry, immunoblotting, and enzyme-linked immunosorbent assay (beta1-adrenergic [beta1-] receptors, muscarinic M2-acetylcholine [M2-] receptors, troponin I, or Na-K ATPase) were measured in 104 patients with DCM. Cardiodepressant antibodies were also determined by ultrasonic echocardiography (UCG) of 18 day old chick embryos after adding the patients' purified immunoglobulin G, and the following clinical features were compared: age, gender, New York Heart Association class, LVEF, neurohumoral factors, arrhythmias, and other antibodies. We also checked the in vitro immunoadsorption effect against these cardiodepressant antibodies. Cardiodepressant antibodies were found in 63% of 104 patients with DCM and had no relation to other clinical parameters, except for some antibodies such as anti beta1-receptor antibodies (81% vs. 52%, P < 0.01), anti-M2-receptor antibodies (83% vs. 48%, P < 0.01), or anti-Na-K-ATPase antibodies (85% vs. 55%, P < 0.01). However, cardiodepressant antibodies were similarly found in patients with and without antibodies against troponin I (56% vs. 64%). The LVEF of chick embryos measured by UCG in the presence of patient serum was improved after in vitro immunoadsorption. The ex vivo system using chick embryos was able to determine cardiodepressant antibodies. By multivariate analysis, antibodies against beta1- or M2-receptors was a predictor of these autoantibodies. PMID- 18387159 TI - Dose determination of cinacalcet hydrochloride in Japanese hemodialysis patients with secondary hyperparathyroidism. AB - Cinacalcet hydrochloride is a calcimimetic agent that activates the calcium sensing receptor on the surface of parathyroid cells and inhibits parathyroid hormone (PTH) secretion. To manage secondary hyperparathyroidism, cinacalcet, which lowers PTH levels without increasing serum calcium, phosphorus and calcium phosphorus product (Ca x P) levels, may provide a new potential therapy. To identify the optimal starting dose of cinacalcet for Japanese hemodialysis patients with secondary hyperparathyroidism, this double-blind, placebo controlled, parallel, dose-finding study was conducted. One hundred and twenty Japanese hemodialysis patients with intact PTH levels greater than or equal to 300 pg/mL were randomized into four groups: placebo, and 12.5, 25 and 50 mg of cinacalcet. The treatment period was three weeks followed by a two-week follow-up observation period. Cinacalcet decreased serum intact PTH levels in a dose dependent manner, and also decreased serum calcium, phosphorus, Ca x P, tartrate resistant acid phosphatase and osteocalcin levels. The treatment with cinacalcet was generally well tolerated in this study. However, the incidence of treatment related adverse events, such as gastrointestinal disorders and hypocalcemia, and the rate of withdrawal from the study due to treatment-related adverse events were higher in the 50 mg dose group than in the other groups. On the basis of both efficacy and safety results, 25 mg has been identified as the optimal starting dose of cinacalcet for Japanese hemodialysis patients with secondary hyperparathyroidism. PMID- 18387160 TI - Comparison of sevelamer hydrochloride with colestimide, administered alone or in combination with calcium carbonate, in patients on hemodialysis. AB - Since hyperphosphatemia in hemodialysis patients can cause secondary hyperparathyroidism and promotes vascular calcification, serum phosphate (Pi) levels must be controlled by phosphate binders. Although sevelamer and colestimide are known as similar non-calcium, non-aluminum phosphate binders in hemodialysis patients, there are no studies that compare the effects of the two agents as either a monotherapy or in combination with calcium carbonate (CaCO3). We randomly allocated 62 hemodialysis patients with hyperphosphatemia to treatment with sevelamer (3.0 g/day) and colestimide (3.0 g/day). During the study, 35 subjects dropped out, leaving 13 in the sevelamer group and 14 in the colestimide group. After a 2-week CaCO3 washout, all subjects received the monotherapy for 4 weeks and then CaCO3 (3.0 g/day) was added for another 4 weeks. Serum corrected calcium levels tended to decrease in both groups during the washout period and monotherapy, but there was no significant difference between the two groups after the addition of CaCO3. Although the calcium x phosphorus product (Ca x P) in the two groups increased during the washout period, there was no significant change or difference between the two groups during monotherapy. However, the addition of CaCO3 significantly reduced serum Pi at Week 8 compared to that at Week 0 in both groups, and significantly lowered Ca x P only in the sevelamer group, but not in the colestimide group(.) In this short-term study, sevelamer and colestimide similarly ameliorated hyperphosphatemia, but the combination of sevelamer and CaCO3 was more effective than colestimide with CaCO3 in controlling the Ca x P product, and it may improve cardiovascular mortality in hemodialysis patients. PMID- 18387161 TI - Platelet (dys)function and plasma plasminogen levels in hemodialysis patients. AB - The factors contributing to platelet dysfunction in hemodialysis patients are still not completely known. We explored whether the fibrinolytic system influences platelet function in hemodialysis patients. We measured standard fibrinolytic parameters and markers of fibrinolysis/coagulation activation, and correlated them to platelet aggregation in 15 hemodialysis patients. Fifteen healthy age-matched volunteers served as controls. Hemodialysis patients had significantly decreased levels of plasminogen (0.76 [0.64-0.86] vs. 0.98 [0.87 1.08] rel, P < 0.001), and increased levels of fibrinogen (4.6 [3.9-5.5] vs. 4.0 [3.4-4.6] g/L, P < 0.05), whereas tissue-type plasminogen activator antigen and plasminogen activator inhibitor (PAI)-1 antigen and PAI activity were comparable to controls. Furthermore, elevated levels of markers of fibrinolysis/coagulation were found in hemodialysis patients: D-dimer (280 [170-460] vs. 135 [120-150] ng/mL, P < 0.01), prothrombin fragments 1 + 2 (1.7 [1.4-1.9] vs. 1.1 [1.0-1.2] nmol/L, P < 0.001), and thrombin-antithrombin complexes (5.2 [4.2-17.7] vs. 0 [0 4.2]microg/L, P < 0.01). The aggregation of platelets (induced by adenosine diphosphate) was slightly impaired in patients compared to controls (72 [43-79] vs. 83 [73-88]%, P = 0.08). Analysis showed that platelet aggregation positively correlated with plasminogen levels (r = 0.48, P < 0.01). No correlation with other fibrinolytic parameters or markers of activation was found. In hemodialysis patients platelet (dys)function appears to be associated with both the fibrinolytic and coagulation systems. We found that platelet aggregation significantly correlates with plasma plasminogen levels. This relation, which has not been hitherto described, seems to be causal and clinically important. Further exploration of this may help us to better understand the mechanisms of platelet dysfunction in hemodialysis patients. PMID- 18387162 TI - Intravenous alfacalcidol once weekly suppresses parathyroid hormone in hemodialysis patients. AB - Management of secondary hyperparathyroidism is difficult because of the interrelationship of parathyroid hormone, calcium and phosphorus. This study was carried out to assess the efficacy of intravenous administration of alfacalcidol once weekly versus twice weekly in patients with severe hyperparathyroidism. Twenty-one hemodialysis patients with intact parathyroid hormone >88 pmol/L were divided into two groups. Eleven patients (Group 1) were given a once-weekly alfacalcidol intravenously for 12 weeks. The starting dose was 4 microg which was increased or decreased by 1 microg per week. Ten patients (Group 2) were given twice-weekly alfacalcidol intravenously for 12 weeks. The starting dose was 2 microg twice weekly which was increased or decreased by 0.5 microg/dose. The dose was increased or decreased according to serum calcium and phosphorus levels. Serum calcium, phosphorus and alkaline phosphatase levels were measured weekly and intact parathyroid hormone every 4 weeks. Intact parathyroid hormone reduced significantly (P = 0.0001) from 128.12 +/- 35.42 pmol/L to 82.93 +/- 65.20 pmol/L and from 113.74 +/- 40.83 pmol/L to 64.24 +/- 35.17 pmol/L after 4 weeks in Groups 1 and 2, respectively. After 4 weeks alkaline phosphatase declined significantly (P = 0.0001) from 146.0 +/- 57.3 IU/L to 116.0 +/- 45.6 IU/L in Group 1 and from 139.0 +/- 45.1 IU/L to 116.6 +/- 38 IU/L in Group 2. There were no significant differences in serum levels of calcium, phosphorous or their product. Interestingly, an adenoma disappeared in one patient from Group 1, and out of two adenomas, one disappeared from another patient in the same group. These results indicate that intravenous alfacalcidol once weekly is safe and effective in suppressing high parathyroid hormone in hemodialysis patients. PMID- 18387163 TI - Persistence of hepatitis B vaccine immunity in hemodialysis patients. AB - Although the efficacy of hepatitis B vaccines in patients undergoing chronic hemodialysis (HD) treatment has been documented, the persistence of immunity in this population remains largely unknown. In this study we evaluated the persistence of hepatitis B vaccine immunity in HD patients. We followed 37 hepatitis B vaccinated HD patients (following a four-dose vaccination schedule of 40 mug injections intramuscularly in the deltoid muscle at 0, 1, 2, and 6 months) for up to one year to evaluate the persistence of immunity (as indicated by serum levels of hepatitis B surface antibody (anti-HBs) equal to or higher than 10 IU/L). One year after vaccination, 18.9% of patients had lost their anti-HBs (transient responders), while 81.1% still had detectable antibodies in the serum (persistent responders). From 81.1% of persistent responders 11.5% and 88.5% were weak and high responders, respectively. There was no significant difference between persistent and transient responders regarding age, sex, or nutritional factors. We did not find any factors that related to maintaining protective levels of anti-HBs in HD patients. It seems that an antibody titer above 100 IU/L following vaccination is necessary in order to maintain that level of antibody one year later. PMID- 18387164 TI - Assessment of plasma antioxidant status in hemodialysis patients. AB - The risk of atherosclerosis and cancer is high in patients on hemodialysis. A breakdown in the natural balance between the activity of the body's antioxidant system and the production of oxidizing agents is suggested to be involved. To investigate the oxidative stress status in Iranian hemodialytic patients, in this study we evaluated plasma vitamin E, malondialdehyde (MDA), reduced glutathione (GSH), and ferric reducing antioxidant power (FRAP) levels in these patients. Twenty-four hemodialytic patients and 24 control subjects (age and sex matched) were included in this study. Each patient was under dialysis, three times per week, four hours in each session. Before and after dialysis, blood was taken for biochemical measurements as well as oxidative stress tests. There was a significant decrease in FRAP and GSH levels after dialysis comparing to before treatment levels. MDA was increased by dialysis and vitamin E levels were less in dialytic patients, both before and after treatment, compared to controls. This study indicates that there is a significant level of oxidative stress in chronic renal patients and this stress is augmented by dialysis. Antioxidant therapy could be considered in these patients. PMID- 18387165 TI - Influence of intravenous drug abuse on native arteriovenous fistula thrombosis in chronic hemodialysis patients. AB - The influence of intravenous drug abuse (IVDA) on native arteriovenous fistula thrombosis (NAT) in chronic hemodialysis patients is unknown. We conducted a retrospective study of 123 chronic hemodialysis patients incarcerated in a male prison in Mid-Taiwan. All patients were dialyzed three times per week, 4 h per session. The development of NAT was compared in patients with a history of IVDA (42, 34.1%) and those without (81, 65.9%). A total of 36 patients experienced one or more NAT episodes in 6 years and 29 (80.6%) of them had a history of IVDA. In these 29 patients, 22 (75.9%) relied on repeated thrombectomy or thrombolysis therapy to maintain patent vascular access. In Kaplan-Meier survival analysis, patients with a history of IVDA were prone to develop NAT (log-rank, P < 0.001). The result of Cox regression suggested that a history of IVDA, independent of diabetes, was associated with the development of NAT. Patients with a history of IVDA, even after IVDA has stopped, were still prone to develop NAT and vascular access infection. PMID- 18387166 TI - Radial augmentation index is related to cardiovascular risk in hemodialysis patients. AB - Cardiovascular accidents related to atherosclerosis are the leading cause of death among hemodialysis patients, which makes continuous monitoring of their cardiovascular status crucial. Recently, a handy device for monitoring the augmentation index (AIx) in the radial artery was introduced in Japan, enabling the use of the AIx in addition to pulse wave velocity (PWV) in the management of hemodialysis patients. In this study the AIx, PWV, abdominal aortic calcification index (ACI), and left ventricular mass index (LVMI) were serially assessed in 108 hemodialysis patients. The radial AIx was monitored using a newly introduced tonometer (HEM-9010AI), and the interrelationships among the measured parameters and their contributions to the risk of cardiovascular accidents were evaluated. The radial AIx was significantly higher in hemodialysis patients than in healthy subjects (N = 50) and was well correlated with risk markers such as LVMI (r = 0.30, P = 0.019) and ACI (r = 0.38, P < 0.001), but not with PWV. Multiregression analysis showed that radial AIx was also significantly associated with LVMI, ACI and blood pressure; PWV was associated with other parameters such as age, blood pressure, and ACI. The AIx and ACI were both significantly increased in patients with cardiovascular complications. Although PWV was strongly increased in the hemodialysis patients, it failed to discriminate between these subgroups of high risk patients. The radial AIx is closely associated with aortic calcification, cardiac hypertrophy, and a history of cardiovascular accidents in hemodialysis patients, and could be a useful marker for management of these patients. PMID- 18387167 TI - Compositional and oxidative analysis in the saliva and serum of predialysis chronic kidney disease patients and end-stage renal failure patients on peritoneal dialysis. AB - Chronic renal failure (CRF) is often treated with peritoneal dialysis, although increased oxidative stress has been reported in such patients. The purpose of the current study was to analyze and compare oxidative stress and other compositional parameters in the saliva, serum and peritoneal dialytic fluid (PDF) of patients with chronic kidney disease (CKD), including predialysis CKD patients and end stage renal disease (ESRD) patients treated with peritoneal dialysis. Twenty three consenting patients participated in the current study. Saliva and serum samples collected from both groups and PDF from the dialysis patients were all examined for uric acid (UA), total antioxidant status, total protein and total albumin. The antioxidant enzyme peroxidase was examined both in saliva and serum, while the antioxidant enzyme superoxide dismutase (SOD) was examined solely in saliva. Various electrolytes were examined. Discrepancies were found between saliva and serum antioxidant status following peritoneal dialysis in ESRD patients. Oxidative stress was enhanced in the saliva but reduced in the serum. Significant changes in both oxidative-related and non-related parameters were demonstrated in saliva, serum and PDF. Salivary lactate dehydrogenase was substantially lower in the dialysis patients (by 92%, P = 0.02), as was the salivary UA concentration (by 22%, P = 0.05) and serum UA concentration (by 20%, P = 0.03). In contrast, salivary peroxidase and SOD were higher by 15% and 35%, respectively (P = 0.01), in these patients. We suggest monitoring salivary UA for assessing the baseline oral oxidative status of CRF and dialyzed patients. PMID- 18387168 TI - Clinical evaluation of pneumonia-associated rhabdomyolysis with acute renal failure. AB - This study aims to evaluate the clinical features, diagnosis, and treatment efficacy in patients with pneumonia-associated rhabdomyolysis and acute renal failure. The subjects included six patients who had presented with rhabdomyolysis and acute renal failure due to bacterial or viral pneumonia on admission to our university hospital and the Yokohama Social Insurance Central Hospital between 2004 and 2005. The causative organisms were identified as Legionella pneumophila (N = 1), Staphylococcus epidermidis (N = 2), Staphylococcus aureus (N = 1), and Unknown (N = 2). For anuric or oliguric patients (N = 4), a blood purification therapy was performed, while conservative therapy was administered to those with a normal urine volume (N = 2). The patient suffering from L. pneumophila pneumonia did not survive, while the other patients regained full kidney function. It is important to identify, evaluate, and treat patients with bacterial or viral pneumonia-associated rhabdomyolysis and acute renal failure. PMID- 18387169 TI - A report of two cases of Kawasaki disease treated with plasma exchange. AB - Kawasaki disease is a generalized vasculitis of unknown etiology that occurs predominantly in infants and young children. It is very important to prevent its cardiovascular manifestations, especially coronary artery lesions. Early treatment with intravenous immunoglobulin reduces cardiovascular sequelae, but some patients do not respond to this treatment, and they have a high incidence of coronary artery lesions. On the other hand, acute heart failure is rare in Kawasaki disease. We report on the cases of two patients with persistent fever and shock even after intravenous immunoglobulin therapy. In both cases, plasma exchange may have reduced the risk of coronary artery lesions and proved effective against acute heart failure with catecholamine-refractory shock; yet the mechanism of this improvement remains unclear. PMID- 18387170 TI - A case report on the role of plasma exchange in the management of a massive amlodipine besylate intoxication. AB - Therapeutic plasma exchange is a procedure used to remove pathologic substances from a patient's blood that has proven useful in some cases of drug overdose. Overdose by calcium channel blocker antihypertensive agents has been shown to be a cause of significant morbidity and can often times prove fatal. These agents cause systemic hypotension by inhibiting cell membrane calcium channels, which leads to a slowing of intracardiac electric conduction with consequent impairment of myocardial function and widespread vasodilation. Shock and metabolic acidosis result from the persistent hypotension. In high doses, calcium channel blocking agents cause insulin resistance. We describe the case of a previously healthy young woman who ingested a massive dose of amlodipine and was treated by therapeutic plasma exchange after non-responsiveness to conventional therapy. The case illustrates the need for utilization of therapeutic plasma exchange in the emergency management of certain cases of severe amlodipine overdose. PMID- 18387171 TI - A case report of refractory warm autoimmune hemolytic anemia treated with plasmapheresis and rituximab. AB - A 30-year-old male presented with severe, warm autoimmune hemolysis 17 months subsequent to a matched, unrelated peripheral hematopoietic stem cell transplant. The patient responded poorly to conventional therapy with steroids and immunoglobulin, prompting the initiation of rituximab. On account of persistent, severe hemolysis, therapeutic plasma exchange was employed as a bridge until the rituximab therapy became effective. Immediately following plasmapheresis, the patient demonstrated clinical improvement followed by attenuation of the hemolysis and improved reticulocytosis. The hemoglobin concentration and reticulocyte index demonstrated further improvement following subsequent doses of rituximab and continued following the cessation of plasmapheresis. This case suggests the utility of plasmapheresis and rituximab in severe, life-threatening cases of warm autoimmune hemolytic anemia refractory to conventional therapy. PMID- 18387172 TI - A case report of recurrent vascular access thrombosis in a hemodialysis patient reveals combined acquired and inherited thrombophilia. AB - Vascular access thrombosis represents a serious and unfortunately common problem in hemodialysis patients. Usually, but not always, this complication can be attributed to low access blood flow. However, there are some patients who experience thrombosis despite a well functioning vascular access. We describe the case of a 31-year-old Caucasian male, who was hemodialyzed via an arteriovenous fistula for two years due to Alport's syndrome. During this time period he had two episodes of vascular access thrombosis that destroyed two arteriovenous fistulas. Both fistulas were functioning well and the thrombosis events took place in days between the hemodialysis sessions. Thrombophilia was suspected and the relative investigation revealed high levels of factor VIII procoagulant, which is frequent in hemodialysis patients, and resistance to activated protein C. Polymerase chain reaction detected that the patient was heterozygous for factor V Leiden, which is quite common in general population. Thereafter, a new arteriovenous fistula was formed and the patient started oral anticoagulation therapy with warfarin. Now, three years after the last arteriovenous fistula formation, the patient is hemodialyzed without vascular access problems. In conclusion, evaluation of the coagulation cascade in hemodialysis patients with recurrent vascular access thrombosis is necessary. PMID- 18387173 TI - The RGS gene loco is essential for male reproductive system differentiation in Drosophila melanogaster. AB - BACKGROUND: The loco gene encodes several different isoforms of a regulator of G protein signalling. These different isoforms of LOCO are part of a pathway enabling cells to respond to external signals. LOCO is known to be required at various developmental stages including neuroblast division, glial cell formation and oogenesis. Less is known about LOCO and its involvement in male development therefore to gain further insight into the role of LOCO in development we carried out a genetic screen and analysed males with reduced fertility. RESULTS: We identified a number of lethal loco mutants and four semi-lethal lines, which generate males with reduced fertility. We have identified a fifth loco transcript and show that it is differentially expressed in developing pupae. We have characterised the expression pattern of all loco transcripts during pupal development in the adult testes, both in wild type and loco mutant strains. In addition we also show that there are various G-protein alpha subunits expressed in the testis all of which may be potential binding partners of LOCO. CONCLUSION: We propose that the male sterility in the new loco mutants result from a failure of accurate morphogenesis of the adult reproductive system during metamorphosis, we propose that this is due to a loss of expression of loco c3. Thus, we conclude that specific isoforms of loco are required for the differentiation of the male gonad and genital disc. PMID- 18387174 TI - A school-based randomized controlled trial to improve physical activity among Iranian high school girls. AB - BACKGROUND: Physical activity (PA) rates decline precipitously during the high school years and are consistently lower among adolescent girls than adolescent boys. Due to cultural barriers, this problem might be exacerbated in female Iranian adolescents. However, little intervention research has been conducted to try to increase PA participation rates with this population. Because PA interventions in schools have the potential to reach many children and adolescents, this study reports on PA intervention research conducted in all female Iranian high schools. METHODS: A randomized controlled trial was conducted to examine the effects of two six-month tailored interventions on potential determinants of PA and PA behavior. Students (N = 161) were randomly allocated to one of three conditions: an intervention based on Pender's Health Promotion model (HP), an intervention based on an integration of the health promotion model and selected constructs from the Transtheoretical model (THP), and a control group (CON). Measures were administered prior to the intervention, at post-intervention and at a six-month follow-up. RESULTS: Repeated measure ANOVAs showed a significant interaction between group and time for perceived benefits, self efficacy, interpersonal norms, social support, behavioral processes, and PA behavior, indicating that both intervention groups significantly improved across the 24-week intervention, whereas the control group did not. Participants in the THP group showed greater use of counter conditioning and stimulus control at post intervention and at follow-up. While there were no significant differences in PA between the HP and CON groups at follow-up, a significant difference was still found between the THP and the CON group. CONCLUSION: This study provides the first evidence of the effectiveness of a PA intervention based on Pender's HP model combined with selected aspects of the TTM on potential determinants to increase PA among Iranian high school girls. PMID- 18387175 TI - Caffeine blocks disruption of blood brain barrier in a rabbit model of Alzheimer's disease. AB - High levels of serum cholesterol and disruptions of the blood brain barrier (BBB) have all been implicated as underlying mechanisms in the pathogenesis of Alzheimer's disease. Results from studies conducted in animals and humans suggest that caffeine might be protective against Alzheimer's disease but by poorly understood mechanisms. Using rabbits fed a cholesterol-enriched diet, we tested our hypothesis that chronic ingestion of caffeine protects against high cholesterol diet-induced disruptions of the BBB. New Zealand rabbits were fed a 2% cholesterol-enriched diet, and 3 mg caffeine was administered daily in drinking water for 12 weeks. Total cholesterol and caffeine concentrations from blood were measured. Olfactory bulbs (and for some studies hippocampus and cerebral cortex as well) were evaluated for BBB leakage, BBB tight junction protein expression levels, activation of astrocytes, and microglia density using histological, immunostaining and immunoblotting techniques. We found that caffeine blocked high cholesterol diet-induced increases in extravasation of IgG and fibrinogen, increases in leakage of Evan's blue dye, decreases in levels of the tight junction proteins occludin and ZO-1, increases in astrocytes activation and microglia density where IgG extravasation was present. Chronic ingestion of caffeine protects against high cholesterol diet-induced increases in disruptions of the BBB, and caffeine and drugs similar to caffeine might be useful in the treatment of Alzheimer's disease. PMID- 18387176 TI - Mosquito larvicidal activities of Solanum villosum berry extract against the dengue vector Stegomyia aegypti. AB - BACKGROUND: Vector control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. Although several plants have been reported for mosquitocidal activity, only a few botanicals have moved from the laboratory to field use, because they are poorly characterized, in most cases active principals are not determined and most of the works are restricted to preliminary screening. Solanum villosum is a common weed distributed in many parts of India with medicinal properties, but the larvicidal activity of this plant has not been reported so far. METHODS: Aqueous and polar/non-polar solvent extract of fresh, mature, green berries of S. villosum was tested against Stegomyia aegypti, a common vector of dengue fever. A phytochemical analysis of chloroform:methanol extract was performed to search for the active toxic ingredient. The lethal concentration was determined (log probit analysis) and compared with Malathion. The chemical nature of the active substance was also evaluated following ultraviolet-visual (UV-Vis) and infrared (IR) analysis. RESULTS: In a 72 hour bioassay experiment with the aqueous extract, the highest mortality was recorded in 0.5% extract. When the mortality of different solvent extracts was compared, the maximum (p < 0.05) mortality was recorded at a concentration of 50 ppm of chloroform:methanol extract (1:1, v/v). The larvicidal activity was lower when compared with the chemical insecticide, Malathion (p < 0.05). Results of regression analysis revealed that the mortality rate (Y) was positively correlated with the period of exposure (X) and the log probit analysis (95% confidence level) recorded lowest value (5.97 ppm) at 72 hours of exposure. Phytochemical analysis of the chlororm:methanol extract reported the presence of many bioactive phytochemicals. Two toxic compounds were detected having Rf = 0.82 (70% and 73.33% mortality in 24 and 48 hours, respectively) and Rf = 0.95 (40% and 50% mortality in 24 and 48 hours, respectively). IR analysis provided preliminary information about the steroidal nature of the active ingredient. CONCLUSION: S. villosum offers promise as potential bio control agent against S. aegypti particularly in its markedly larvicidal effect. The extract or isolated bioactive phytochemical could be used in stagnant water bodies for the control of mosquitoes acting as vector for many communicable diseases. PMID- 18387177 TI - Killing cancer cells by targeted drug-carrying phage nanomedicines. AB - BACKGROUND: Systemic administration of chemotherapeutic agents, in addition to its anti-tumor benefits, results in indiscriminate drug distribution and severe toxicity. This shortcoming may be overcome by targeted drug-carrying platforms that ferry the drug to the tumor site while limiting exposure to non-target tissues and organs. RESULTS: We present a new form of targeted anti-cancer therapy in the form of targeted drug-carrying phage nanoparticles. Our approach is based on genetically-modified and chemically manipulated filamentous bacteriophages. The genetic manipulation endows the phages with the ability to display a host-specificity-conferring ligand. The phages are loaded with a large payload of a cytotoxic drug by chemical conjugation. In the presented examples we used anti ErbB2 and anti ERGR antibodies as targeting moieties, the drug hygromycin conjugated to the phages by a covalent amide bond, or the drug doxorubicin conjugated to genetically-engineered cathepsin-B sites on the phage coat. We show that targeting of phage nanomedicines via specific antibodies to receptors on cancer cell membranes results in endocytosis, intracellular degradation, and drug release, resulting in growth inhibition of the target cells in vitro with a potentiation factor of >1000 over the corresponding free drugs. CONCLUSION: The results of the proof-of concept study presented here reveal important features regarding the potential of filamentous phages to serve as drug delivery platform, on the affect of drug solubility or hydrophobicity on the target specificity of the platform and on the effect of drug release mechanism on the potency of the platform. These results define targeted drug-carrying filamentous phage nanoparticles as a unique type of antibody-drug conjugates. PMID- 18387179 TI - A case of spotted fever group rickettsiosis imported into the United Kingdom and treated with ciprofloxacin: a case report. AB - INTRODUCTION: Spotted fever group rickettsioses are an interesting group of infections, which are increasing in incidence worldwide. CASE PRESENTATION: Here we describe an imported case to the United Kingdom occurring in a patient who had recently visited Kruger National Park in South Africa - a highly endemic area for Rickettsia infections. Initial treatment with doxycycline failed but the patient made a prompt recovery after commencement of ciprofloxacin. CONCLUSION: This finding raises the possibility that there are resistant strains of Rickettsia present. PMID- 18387178 TI - Outbreak of haemolytic uraemic syndrome in Norway caused by stx2-positive Escherichia coli O103:H25 traced to cured mutton sausages. AB - BACKGROUND: On 20-21 February 2006, six cases of diarrhoea-associated haemolytic uraemic syndrome (HUS) were reported by paediatricians to the Norwegian Institute of Public Health. We initiated an investigation to identify the etiologic agent and determine the source of the outbreak in order to implement control measures. METHODS: A case was defined as a child with diarrhoea-associated HUS or any person with an infection with the outbreak strain of E. coli O103 (defined by the multi-locus variable number tandem repeats analysis (MLVA) profile) both with illness onset after January 1st 2006 in Norway. After initial hypotheses generating interviews, we performed a case-control study with the first fifteen cases and three controls for each case matched by age, sex and municipality. Suspected food items were sampled, and any E. coli O103 strains were typed by MLVA. RESULTS: Between 20 February and 6 April 2006, 17 cases were identified, of which 10 children developed HUS, including one fatal case. After pilot interviews, a matched case-control study was performed indicating an association between a traditional cured sausage (odds ratio 19.4 (95% CI: 2.4-156)) and STEC infection. E. coli O103:H25 identical to the outbreak strain defined by MLVA profile was found in the product and traced back to contaminated mutton. CONCLUSION: We report an outbreak caused by a rare STEC variant (O103:H25, stx2 positive). More than half of the diagnosed patients developed HUS, indicating that the causative organism is particularly virulent. Small ruminants continue to be important reservoirs for human-pathogen STEC. Improved slaughtering hygiene and good manufacturing practices for cured sausage products are needed to minimise the possibility of STEC surviving through the entire sausage production process. PMID- 18387180 TI - Can changes in malaria transmission intensity explain prolonged protection and contribute to high protective efficacy of intermittent preventive treatment for malaria in infants? AB - BACKGROUND: Intermittent preventive (or presumptive) treatment of infants (IPTi), the administration of a curative anti-malarial dose to infants whether or not they are known to be infected, is being considered as a new strategy for malaria control. Five of the six trials using sulphadoxine-pyrimethamine (SP) for IPTi showed protective efficacies (PEs) against clinical malaria ranging from 20.1 - 33.3% whilst one, the Ifakara study, showed a protective efficacy of 58.6%. MATERIALS AND METHODS: The possible mechanisms that could explain the differences in the reported PE of IPTi were examined by comparing output from a mathematical model to data from the six published IPTi trials. RESULTS: Under stable transmission, the PE of IPTi predicted by the model was comparable with the observed PEs in all but the Ifakara study (ratio of the mean predicted PE to that observed was 1.02, range 0.39 - 1.59). When a reduction in the incidence of infection during the study was included in the model, the predicted PE of IPTi increased and extended into the second year of life, as observed in the Ifakara study. CONCLUSION: A decrease in malaria transmission during the study period may explain part of the difference in observed PEs of IPTi between sites and the extended period of protection into the second year of life observed in the Ifakara study. This finding of continued benefit of interventions in settings of decreasing transmission may explain why rebound of clinical malaria was absent in the large scale trials of insecticide-treated bed nets. PMID- 18387181 TI - Appropriateness of antibiotic treatment in intravenous drug users, a retrospective analysis. AB - BACKGROUND: Infectious disease is often the reason for intravenous drug users being seen in a clinical setting. The objective of this study was to evaluate the appropriateness of treatment and outcomes for this patient population in a hospital setting. METHODS: Retrospective study of all intravenous drug users hospitalized for treatment of infectious diseases and seen by infectious diseases specialists 1/2001-12/2006 at a university hospital. Treatment was administered according to guidelines when possible or to alternative treatment program in case of patients for whom adherence to standard protocols was not possible. Outcomes were defined with respect to appropriateness of treatment, hospital readmission, relapse and mortality rates. For statistical analysis adjustment for multiple hospitalizations of individual patients was made by using a generalized estimating equation. RESULTS: The total number of hospitalizations for infectious diseases was 344 among 216 intravenous drug users. Skin and soft tissue infections (n = 129, 37.5% of hospitalizations), pneumonia (n = 75, 21.8%) and endocarditis (n = 54, 15.7%) were most prevalent. Multiple infections were present in 25%. Treatment was according to standard guidelines for 78.5%, according to an alternative recommended program for 11.3%, and not according to guidelines or by the infectious diseases specialist advice for 10.2% of hospitalizations. Psychiatric disorders had a significant negative impact on compliance (compliance problems in 19.8% of hospitalizations) in multiple logistic regression analysis (OR = 2.4, CI 1.1-5.1, p = 0.03). The overall readmission rate and relapse rate within 30 days was 13.7% and 3.8%, respectively. Both non-compliant patient behavior (OR = 3.7, CI 1.3-10.8, p = 0.02) and non-adherence to treatment guidelines (OR = 3.3, CI 1.1-9.7, p = 0.03) were associated with a significant increase in the relapse rate in univariate analysis. In 590 person-years of follow-up, 24.6% of the patients died: 6.4% died during hospitalization (1.2% infection-related) and 13.6% of patients died after discharge. CONCLUSION: Appropriate antibiotic therapy according to standard guidelines in hospitalized intravenous drug users is generally practicable and successful. In a minority alternative treatments may be indicated, although associated with a higher risk of relapse. PMID- 18387182 TI - Characterization and site-directed mutagenesis of Wzb, an O-phosphatase from Lactobacillus rhamnosus. AB - BACKGROUND: Reversible phosphorylation events within a polymerisation complex have been proposed to modulate capsular polysaccharide synthesis in Streptococcus pneumoniae. Similar phosphatase and kinase genes are present in the exopolysaccharide (EPS) biosynthesis loci of numerous lactic acid bacteria genomes. RESULTS: The protein sequence deduced from the wzb gene in Lactobacillus rhamnosus ATCC 9595 reveals four motifs of the polymerase and histidinol phosphatase (PHP) superfamily of prokaryotic O-phosphatases. Native and modified His-tag fusion Wzb proteins were purified from Escherichia coli cultures. Extracts showed phosphatase activity towards tyrosine-containing peptides. The purified fusion protein Wzb was active on p-nitrophenyl-phosphate (pNPP), with an optimal activity in presence of bovine serum albumin (BSA 1%) at pH 7.3 and a temperature of 75 degrees C. At 50 degrees C, residual activity decreased to 10 %. Copper ions were essential for phosphatase activity, which was significantly increased by addition of cobalt. Mutated fusion Wzb proteins exhibited reduced phosphatase activity on p-nitrophenyl-phosphate. However, one variant (C6S) showed close to 20% increase in phosphatase activity. CONCLUSION: These characteristics reveal significant differences with the manganese-dependent CpsB protein tyrosine phosphatase described for Streptococcus pneumoniae as well as with the polysaccharide-related phosphatases of Gram negative bacteria. PMID- 18387183 TI - Sex-specific incidence and temporal trends in solid tumours in young people from Northern England, 1968-2005. AB - BACKGROUND: This study examined sex-specific patterns and temporal trends in the incidence of solid tumours in the Northern Region of England from 1968 to 2005. This updates earlier analyses from the region where sex was not considered in depth. Sex-specific analyses were carried out to determine whether sex differences might provide clues to aetiology. METHODS: Details of 3576 cases, aged 0-24 years, were obtained from a specialist population-based cancer registry. There were 1843 males (886 aged 0-14 years and 957 aged 15-24 years) and 1733 females (791 aged 0-14 years and 942 aged 15-24 years). Age-standardized incidence rates (per million population) were calculated. Linear regression was used to analyze temporal trends in incidence and annual percentage changes were estimated. Analyses were stratified by sex and by age-group. RESULTS: There were marked differences in incidence patterns and trends between males and females and also between age-groups. For males central nervous system (CNS) tumours formed the largest proportion of under-15 cases and germ cell tumours was the largest group in the 15-24's, whilst for females CNS tumours dominated in the under-15's and carcinomas in the older group. For 0-14 year olds there were male-specific increases in the incidence of rhabdomyosarcoma (2.4% per annum; 95% CI: 0.2% 4.5%) and non-melanotic skin cancer (9.6%; 95% CI: 0.0%-19.2%) and female specific increases for sympathetic nervous system tumours (2.2%; 95% CI: 0.4% 3.9%), gonadal germ cell tumours (8.6%; 95% CI: 4.3%-12.9%) and non-gonadal germ cell tumours (5.4%; 95% CI: 2.8%-7.9%). For 15-24 year olds, there were male specific increases in gonadal germ cell tumours (1.9%; 95% CI: 0.3%-3.4%), non gonadal germ cell tumours (4.4%; 95% CI: 1.1%-7.7%) and non-melanotic skin cancer (4.7%; 95% CI: 0.5%-8.9%) and female-specific increases for osteosarcoma (3.5%; 95% CI: 0.5%-6.5%), thyroid cancer (2.8%; 95% CI: 0.1%-5.6%) and melanoma (4.6%; 95% CI: 2.2%-7.1%). CONCLUSION: This study has highlighted notable differences between the sexes in incidence patterns and trends for solid tumours. Some of these sex-specific differences could have been obscured if males and females had been analysed together. Furthermore, they suggest aetiological differences or differential susceptibility to environmental factors between males and females. PMID- 18387184 TI - Effects of intensive home visiting programs for older people with poor health status: a systematic review. AB - BACKGROUND: Home visiting programs have been developed aimed at improving the health and independent functioning of older people. Also, they intend to reduce hospital and nursing home admission and associated cost. A substantial number of studies have examined the effects of preventive home visiting programs on older people living in the community; the findings have been inconsistent. The objective of this review was to assess the effectiveness of intensive home visiting programs targeting older people with poor health or otherwise with functional impairments. METHODS: A search for literature was based on included trials from four reviews on the effectiveness of home visits published after 2000 and on a database search of Cinahl, the Cochrane Central Register of Controlled Trials, Embase, Medline and PsycINFO from 2001 onwards. We also manually searched reference lists from potentially relevant papers. Randomized controlled trials were included assessing the effectiveness of intervention programs consisting of at least four home visits per year, an intervention duration of 12 months or more, and targeting older people (aged 65 years and over) with poor health. Two reviewers independently abstracted data from full papers on program characteristics and outcome measures; they also evaluated the methodological quality. RESULTS: The search identified 844 abstracts; eight papers met the inclusion criteria. Seven trials were of sufficient methodological quality; none of the trials showed a significant favorable effect for the main analysis comparing the intervention group with the control group on mortality, health status, service use or cost. The inclusion of less-intensive intervention programs for frail older persons would not have exerted a great influence on the findings of our review. CONCLUSION: We conclude that home visiting programs appear not to be beneficial for older people with poor health within the health care setting of Western countries. PMID- 18387185 TI - Design of the Pacemaker REmote Follow-up Evaluation and Review (PREFER) trial to assess the clinical value of the remote pacemaker interrogation in the management of pacemaker patients. AB - BACKGROUND: Although pacemakers are primarily used for the treatment of bradycardia, diagnostic data available in current pacemakers allow them to be also used as sophisticated, continuous monitoring devices. Easy access to these stored data may assist clinicians in making diagnostic and therapeutic decisions sooner, thus avoiding potential long-term sequelae due to untreated clinical disorders. Internet-based remote device interrogation systems provide clinicians with frequent and complete access to stored data in pacemakers. In addition to monitoring device function, remote monitors may be a helpful tool in assisting physicians in the management of common arrhythmia disorders. METHODS: The Pacemaker REmote Follow-up Evaluation and Review (PREFER) trial is a prospective, randomized, parallel, unblinded, multicenter, open label clinical trial to determine the utility of remote pacemaker interrogation in the earlier diagnosis of clinically actionable events compared to the existing practice of transtelephonic monitoring. There have been 980 patients enrolled and randomized to receive pacemaker follow up with either remote interrogation using the Medtronic CareLink(R) Network (CareLink) versus the conventional method of transtelephonic monitoring (TTM) in addition to periodic in-person interrogation and programming evaluations. The purpose of this manuscript is to describe the design of the PREFER trial. The results, to be presented separately, will characterize the number of clinically actionable events as a result of pacemaker follow-up using remote interrogation instead of TTM. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00294645. PMID- 18387186 TI - Metals and kidney markers in adult offspring of endemic nephropathy patients and controls: a two-year follow-up study. AB - BACKGROUND: The etiology of Balkan Endemic Nephropathy, (BEN), a tubulointerstitial kidney disease, is unknown. Although this disease is endemic in rural areas of Bosnia, Bulgaria, Croatia, Romania, and Serbia, similar manifestations are reported to occur in other regions, for instance Tunisia and Sri Lanka. A number of explanations have been stated including lignites, aristolochic acid, ochratoxin A, metals, and metalloids. Etiologic claims are often based on one or a few studies without sound scientific evidence. In this systematic study, we tested whether exposures to metals (cadmium and lead) and metalloids (arsenic and selenium) are related to Balkan Endemic Nephropathy. METHODS: In 2003/04 we recruited 102 adults whose parents had BEN and who resided in one of three communities (Vratza, Bistretz, or Beli Izvor, Bulgaria). A control group comprised of 99 adults having non-BEN hospitalized parents was enrolled in the study during the same time. We conducted face-to-face interviews, ultrasound kidney measurements, and determined kidney function in two consecutive investigations (2003/04 and 2004/05). Metals and metalloids were measured in urine and blood samples. To assess the agreement between these consecutive measurements, we calculated intraclass correlation coefficients. Repeated measurement data were analyzed using mixed models. RESULTS: We found that cadmium and arsenic were associated with neither kidney size nor function. Lead had a significant but negligible effect on creatinine clearance. Selenium showed a weak but significant negative association with two of the four kidney parameters, namely creatinine clearance and beta2-microglobulin. It was positively related to kidney length. These associations were not restricted to the offspring of BEN patients. Adding credence to these findings are reports showing comparable kidney effects in animals exposed to selenium. CONCLUSION: The findings of this 2-year follow-up study indicate that metals and metalloids do not play a role in the etiology of Balkan Endemic Nephropathy. Against the assumption in the literature, selenium was not protective but a risk factor. Since comparable associations were observed in animals, future studies are needed to explore whether selenium may have adverse renal effects in humans. PMID- 18387187 TI - The impact of consent on observational research: a comparison of outcomes from consenters and non consenters to an observational study. AB - BACKGROUND: Public health benefits from research often rely on the use of data from personal medical records. When neither patient consent nor anonymisation is possible, the case for accessing such records for research purposes depends on an assessment of the probabilities of public benefit and individual harm. METHODS: In the late 1990s, we carried out an observational study which compared the care given to affluent and deprived women with breast cancer. Patient consent was not required at that time for review of medical records, but was obtained later in the process prior to participation in the questionnaire study. We have re analysed our original results to compare the whole sample with those who later provided consent. RESULTS: Two important findings emerged from the re-analysis of our data which if presented initially would have resulted in insufficient and inaccurate reporting. Firstly, the reduced dataset contains no information about women presenting with locally advanced or metastatic cancer and we would have been unable to demonstrate one of our initial key findings: namely a larger number of such women in the deprived group. Secondly, our re-analysis of the consented women shows that significantly more women from deprived areas (51 v 31%, p = 0.018) received radiotherapy compared to women from more affluent areas. Previously published data from the entire sample demonstrated no difference in radiotherapy treatment between the affluent and deprived groups. CONCLUSION: The risk benefit assessment made regarding the use of medical records without consent should include the benefits of obtaining research evidence based on 100% of the population and the possibility of inappropriate or insufficient findings if research is confined to consented populations. PMID- 18387188 TI - Validation and extension of an empirical Bayes method for SNP calling on Affymetrix microarrays. AB - Multiple algorithms have been developed for the purpose of calling single nucleotide polymorphisms (SNPs) from Affymetrix microarrays. We extend and validate the algorithm CRLMM, which incorporates HapMap information within an empirical Bayes framework. We find CRLMM to be more accurate than the Affymetrix default programs (BRLMM and Birdseed). Also, we tie our call confidence metric to percent accuracy. We intend that our validation datasets and methods, refered to as SNPaffycomp, serve as standard benchmarks for future SNP calling algorithms. PMID- 18387189 TI - Geocoding accuracy and the recovery of relationships between environmental exposures and health. AB - BACKGROUND: This research develops methods for determining the effect of geocoding quality on relationships between environmental exposures and health. The likelihood of detecting an existing relationship - statistical power - between measures of environmental exposures and health depends not only on the strength of the relationship but also on the level of positional accuracy and completeness of the geocodes from which the measures of environmental exposure are made. This paper summarizes the results of simulation studies conducted to examine the impact of inaccuracies of geocoded addresses generated by three types of geocoding processes: a) addresses located on orthophoto maps, b) addresses matched to TIGER files (U.S Census or their derivative street files); and, c) addresses from E-911 geocodes (developed by local authorities for emergency dispatch purposes). RESULTS: The simulated odds of disease using exposures modelled from the highest quality geocodes could be sufficiently recovered using other, more commonly used, geocoding processes such as TIGER and E-911; however, the strength of the odds relationship between disease exposures modelled at geocodes generally declined with decreasing geocoding accuracy. CONCLUSION: Although these specific results cannot be generalized to new situations, the methods used to determine the sensitivity of results can be used in new situations. Estimated measures of positional accuracy must be used in the interpretation of results of analyses that investigate relationships between health outcomes and exposures measured at residential locations. Analyses similar to those employed in this paper can be used to validate interpretation of results from empirical analyses that use geocoded locations with estimated measures of positional accuracy. PMID- 18387190 TI - The virtual haptic back: a simulation for training in palpatory diagnosis. AB - BACKGROUND: Models and simulations are finding increased roles in medical education. The Virtual Haptic Back (VHB) is a virtual reality simulation of the mechanical properties of the human back designed as an aid to teaching clinical palpatory diagnosis. METHODS: Eighty-nine first year medical students of the Ohio University College of Osteopathic Medicine carried out six, 15-minute practice sessions with the VHB, plus tests before and after the sessions in order to monitor progress in identifying regions of simulated abnormal tissue compliance. Students palpated with two digits, fingers or thumbs, by placing them in gimbaled thimbles at the ends of PHANToM 3.0(R) haptic interface arms. The interface simulated the contours and compliance of the back surface by the action of electric motors. The motors limited the compression of the virtual tissues induced by the palpating fingers, by generating counterforces. Users could see the position of their fingers with respect to the back on a video monitor just behind the plane of the haptic back. The abnormal region varied randomly among 12 locations between trials. During the practice sessions student users received immediate feedback following each trial, indicating either a correct choice or the actual location of the abnormality if an incorrect choice had been made. This allowed the user to feel the actual abnormality before going on to the next trial. Changes in accuracy, speed and Weber fraction across practice sessions were analyzed using a repeated measures analysis of variance. RESULTS: Students improved in accuracy and speed of diagnosis with practice. The smallest difference in simulated tissue compliance users were able to detect improved from 28% (SD = 9.5%) to 14% (SD = 4.4%) during the practice sessions while average detection time decreased from 39 (SD = 19.8) to 17 (SD = 11.7) seconds. When asked in anonymous evaluation questionnaires if they judged the VHB practice to be helpful to them in the clinical palpation and manual medicine laboratory, 41% said yes, 51% said maybe, and 8% said no. CONCLUSION: The VHB has potential value as a teaching aid for students in the initial phases of learning palpatory diagnosis. PMID- 18387191 TI - Fluorodeoxyglucose-positron emission tomography/computed tomography in the staging and evaluation of treatment response in a patient with Castleman's disease: a case report. AB - INTRODUCTION: Castleman's disease is a rare lymphatic polyclonal disorder that is characterised by unicentric or multicentric lymph node hyperplasia and non specific symptoms and signs including fever, asthenia, weight loss, enlarged liver and abnormally high blood levels of antibodies. CASE PRESENTATION: We present the case of a 74-year-old man with Castleman's disease. The disease was detected with a contrast-enhanced computed tomography (CT) scan and a fluorodeoxyglucose (FDG)-positron emission tomography (PET)/CT study; diagnosis was made with histopathology. After treatment with surgical excision followed by chemotherapy, the disease response was evaluated using both diagnostic techniques. However, only the PET study was able to identify the spread of the disease to the abdominal lymph nodes, which were both enlarged and normal size, and, after treatment, to evaluate the disease response. CONCLUSION: Based on the results of previous case reports and on those of the present study, it seems that Castleman's disease has a high glucose metabolic activity. Therefore, the use of PET can be considered appropriate in order to stage or restage the disease and to evaluate the response of the disease to treatment. PMID- 18387192 TI - A pathway sensor for genome-wide screens of intracellular proteolytic cleavage. AB - Protein cleavage is a central event in many regulated biological processes. We describe a system for detecting intracellular proteolysis based on non conventional secretion of Gaussia luciferase (GLUC). GLUC exits the cell without benefit of a secretory leader peptide, but can be anchored in the cell by fusion to beta-actin. By including protease cleavage sites between GLUC and beta-actin, proteolytic cleavage can be detected. Using this assay, we have identified regulators of autophagy, apoptosis and beta-actin cleavage. PMID- 18387194 TI - Thyroid cancer causing obstruction of the great veins in the neck. AB - BACKGROUND AND AIMS: To report our experience and review the literature of thyroid cancer obstructing the great veins in the neck, highlighting clinical aspects and response to treatment. METHODS: Clinical data were collected from the thyroid cancer register and from follow-up clinic visits of patients referred to the Thyroid Unit at the Royal Marsden Hospital. A Medline literature search was conducted between 1980 and 2007. RESULTS: Of 1448 patients with thyroid cancer on our cancer register and treated in our unit over the last 60 years, we identified five patients, four women and one man, aged 43 - 81 years with a median follow up of 28 (24-78) months in whom tumour had occluded the great veins in the neck. All patients underwent total thyroidectomy and all subsequently received ablative 131I with the exception of patient 3 whose post-operative isotope scan shown no significant 131I uptake. External beam radiotherapy to the neck and upper mediastinum was used for residual disease control in the 5 patients. The median survival was 28 months and the disease-free survival was 24 months. One patient remains asymptomatic but with disease 53 months after initial presentation. Survival in this small series is significantly better than that previously reported for this condition. CONCLUSION: A multimodality therapeutic approach comprising surgery, radioiodine and external beam radiotherapy may give the best results for patients in whom thyroid cancer is occluding the great veins. PMID- 18387193 TI - Spatial distribution of transcript changes in the maize primary root elongation zone at low water potential. AB - BACKGROUND: Previous work showed that the maize primary root adapts to low Psiw ( 1.6 MPa) by maintaining longitudinal expansion in the apical 3 mm (region 1), whereas in the adjacent 4 mm (region 2) longitudinal expansion reaches a maximum in well-watered roots but is progressively inhibited at low Psiw. To identify mechanisms that determine these responses to low Psiw, transcript expression was profiled in these regions of water-stressed and well-watered roots. In addition, comparison between region 2 of water-stressed roots and the zone of growth deceleration in well-watered roots (region 3) distinguished stress-responsive genes in region 2 from those involved in cell maturation. RESULTS: Responses of gene expression to water stress in regions 1 and 2 were largely distinct. The largest functional categories of differentially expressed transcripts were reactive oxygen species and carbon metabolism in region 1, and membrane transport in region 2. Transcripts controlling sucrose hydrolysis distinguished well watered and water-stressed states (invertase vs. sucrose synthase), and changes in expression of transcripts for starch synthesis indicated further alteration in carbon metabolism under water deficit. A role for inositols in the stress response was suggested, as was control of proline metabolism. Increased expression of transcripts for wall-loosening proteins in region 1, and for elements of ABA and ethylene signaling were also indicated in the response to water deficit. CONCLUSION: The analysis indicates that fundamentally different signaling and metabolic response mechanisms are involved in the response to water stress in different regions of the maize primary root elongation zone. PMID- 18387195 TI - A new classification system for bacterial Rieske non-heme iron aromatic ring hydroxylating oxygenases. AB - BACKGROUND: Rieske non-heme iron aromatic ring-hydroxylating oxygenases (RHOs) are multi-component enzyme systems that are remarkably diverse in bacteria isolated from diverse habitats. Since the first classification in 1990, there has been a need to devise a new classification scheme for these enzymes because many RHOs have been discovered, which do not belong to any group in the previous classification. Here, we present a scheme for classification of RHOs reflecting new sequence information and interactions between RHO enzyme components. RESULT: We have analyzed a total of 130 RHO enzymes in which 25 well-characterized RHO enzymes were used as standards to test our hypothesis for the proposed classification system. From the sequence analysis of electron transport chain (ETC) components of the standard RHOs, we extracted classification keys that reflect not only the phylogenetic affiliation within each component but also relationship among components. Oxygenase components of standard RHOs were phylogenetically classified into 10 groups with the classification keys derived from ETC components. This phylogenetic classification scheme was converted to a new systematic classification consisting of 5 distinct types. The new classification system was statistically examined to justify its stability. Type I represents two-component RHO systems that consist of an oxygenase and an FNRC type reductase. Type II contains other two-component RHO systems that consist of an oxygenase and an FNRN-type reductase. Type III represents a group of three component RHO systems that consist of an oxygenase, a [2Fe-2S]-type ferredoxin and an FNRN-type reductase. Type IV represents another three-component systems that consist of oxygenase, [2Fe-2S]-type ferredoxin and GR-type reductase. Type V represents another different three-component systems that consist of an oxygenase, a [3Fe-4S]-type ferredoxin and a GR-type reductase. CONCLUSION: The new classification system provides the following features. First, the new classification system analyzes RHO enzymes as a whole. RwithSecond, the new classification system is not static but responds dynamically to the growing pool of RHO enzymes. Third, our classification can be applied reliably to the classification of incomplete RHOs. Fourth, the classification has direct applicability to experimental work. Fifth, the system provides new insights into the evolution of RHO systems based on enzyme interaction. PMID- 18387196 TI - An integrated genetic, genomic and systems approach defines gene networks regulated by the interaction of light and carbon signaling pathways in Arabidopsis. AB - BACKGROUND: Light and carbon are two important interacting signals affecting plant growth and development. The mechanism(s) and/or genes involved in sensing and/or mediating the signaling pathways involving these interactions are unknown. This study integrates genetic, genomic and systems approaches to identify a genetically perturbed gene network that is regulated by the interaction of carbon and light signaling in Arabidopsis. RESULTS: Carbon and light insensitive (cli) mutants were isolated. Microarray data from cli186 is analyzed to identify the genes, biological processes and gene networks affected by the integration of light and carbon pathways. Analysis of this data reveals 966 genes regulated by light and/or carbon signaling in wild-type. In cli186, 216 of these light/carbon regulated genes are misregulated in response to light and/or carbon treatments where 78% are misregulated in response to light and carbon interactions. Analysis of the gene lists show that genes in the biological processes "energy" and "metabolism" are over-represented among the 966 genes regulated by carbon and/or light in wild-type, and the 216 misregulated genes in cli186. To understand connections among carbon and/or light regulated genes in wild-type and the misregulated genes in cli186, the microarray data is interpreted in the context of metabolic and regulatory networks. The network created from the 966 light/carbon regulated genes in wild-type, reveals that cli186 is affected in the light and/or carbon regulation of a network of 60 connected genes, including six transcription factors. One transcription factor, HAT22 appears to be a regulatory "hub" in the cli186 network as it shows regulatory connections linking a metabolic network of genes involved in "amino acid metabolism", "C compound/carbohydrate metabolism" and "glycolysis/gluconeogenesis". CONCLUSION: The global misregulation of gene networks controlled by light and carbon signaling in cli186 indicates that it represents one of the first Arabidopsis mutants isolated that is specifically disrupted in the integration of both carbon and light signals to control the regulation of metabolic, developmental and regulatory genes. The network analysis of misregulated genes suggests that CLI186 acts to integrate light and carbon signaling interactions and is a master regulator connecting the regulation of a host of downstream metabolic and regulatory processes. PMID- 18387198 TI - CUDA compatible GPU cards as efficient hardware accelerators for Smith-Waterman sequence alignment. AB - BACKGROUND: Searching for similarities in protein and DNA databases has become a routine procedure in Molecular Biology. The Smith-Waterman algorithm has been available for more than 25 years. It is based on a dynamic programming approach that explores all the possible alignments between two sequences; as a result it returns the optimal local alignment. Unfortunately, the computational cost is very high, requiring a number of operations proportional to the product of the length of two sequences. Furthermore, the exponential growth of protein and DNA databases makes the Smith-Waterman algorithm unrealistic for searching similarities in large sets of sequences. For these reasons heuristic approaches such as those implemented in FASTA and BLAST tend to be preferred, allowing faster execution times at the cost of reduced sensitivity. The main motivation of our work is to exploit the huge computational power of commonly available graphic cards, to develop high performance solutions for sequence alignment. RESULTS: In this paper we present what we believe is the fastest solution of the exact Smith Waterman algorithm running on commodity hardware. It is implemented in the recently released CUDA programming environment by NVidia. CUDA allows direct access to the hardware primitives of the last-generation Graphics Processing Units (GPU) G80. Speeds of more than 3.5 GCUPS (Giga Cell Updates Per Second) are achieved on a workstation running two GeForce 8800 GTX. Exhaustive tests have been done to compare our implementation to SSEARCH and BLAST, running on a 3 GHz Intel Pentium IV processor. Our solution was also compared to a recently published GPU implementation and to a Single Instruction Multiple Data (SIMD) solution. These tests show that our implementation performs from 2 to 30 times faster than any other previous attempt available on commodity hardware. CONCLUSIONS: The results show that graphic cards are now sufficiently advanced to be used as efficient hardware accelerators for sequence alignment. Their performance is better than any alternative available on commodity hardware platforms. The solution presented in this paper allows large scale alignments to be performed at low cost, using the exact Smith-Waterman algorithm instead of the largely adopted heuristic approaches. PMID- 18387199 TI - The MoVIN server for the analysis of protein interaction networks. AB - BACKGROUND: Protein-protein interactions are at the basis of most cellular processes and crucial for many bio-technological applications. During the last few years the development of high-throughput technologies has produced several large-scale protein-protein interaction data sets for various organisms. It is important to develop tools for dissecting their content and analyse the information they embed by data-integration and computational methods. RESULTS: Interactions can be mediated by the presence of specific features, such as motifs, surface patches and domains. The co-occurrence of these features on proteins interacting with the same protein can indicate mutually exclusive interactions and, therefore, can be used for inferring the involvement of the proteins in common biological processes. We present here a publicly available server that allows the user to investigate protein interaction data in light of other biological information, such as their sequences, presence of specific domains, process and component ontologies. The server can be effectively used to construct a high-confidence set of mutually exclusive interactions by identifying similar features in groups of proteins sharing a common interaction partner. As an example, we describe here the identification of common motifs, function, cellular localization and domains in different datasets of yeast interactions. CONCLUSIONS: The server can be used to analyse user-supplied datasets, it contains pre-processed data for four yeast Protein Protein interaction datasets and the results of their statistical analysis. These show that the presence of common motifs in proteins interacting with the same partner is a valuable source of information, it can be used to investigate the properties of the interacting proteins and provides information that can be effectively integrated with other sources. As more experimental interaction data become available, this tool will become more and more useful to gain a more detailed picture of the interactome. PMID- 18387200 TI - Time-course analysis of genome-wide gene expression data from hormone-responsive human breast cancer cells. AB - BACKGROUND: Microarray experiments enable simultaneous measurement of the expression levels of virtually all transcripts present in cells, thereby providing a 'molecular picture' of the cell state. On the other hand, the genomic responses to a pharmacological or hormonal stimulus are dynamic molecular processes, where time influences gene activity and expression. The potential use of the statistical analysis of microarray data in time series has not been fully exploited so far, due to the fact that only few methods are available which take into proper account temporal relationships between samples. RESULTS: We compared here four different methods to analyze data derived from a time course mRNA expression profiling experiment which consisted in the study of the effects of estrogen on hormone-responsive human breast cancer cells. Gene expression was monitored with the innovative Illumina BeadArray platform, which includes an average of 30-40 replicates for each probe sequence randomly distributed on the chip surface. We present and discuss the results obtained by applying to these datasets different statistical methods for serial gene expression analysis. The influence of the normalization algorithm applied on data and of different parameter or threshold choices for the selection of differentially expressed transcripts has also been evaluated. In most cases, the selection was found fairly robust with respect to changes in parameters and type of normalization. We then identified which genes showed an expression profile significantly affected by the hormonal treatment over time. The final list of differentially expressed genes underwent cluster analysis of functional type, to identify groups of genes with similar regulation dynamics. CONCLUSIONS: Several methods for processing time series gene expression data are presented, including evaluation of benefits and drawbacks of the different methods applied. The resulting protocol for data analysis was applied to characterization of the gene expression changes induced by estrogen in human breast cancer ZR-75.1 cells over an entire cell cycle. PMID- 18387201 TI - SNPLims: a data management system for genome wide association studies. AB - BACKGROUND: Recent progresses in genotyping technologies allow the generation high-density genetic maps using hundreds of thousands of genetic markers for each DNA sample. The availability of this large amount of genotypic data facilitates the whole genome search for genetic basis of diseases. We need a suitable information management system to efficiently manage the data flow produced by whole genome genotyping and to make it available for further analyses. RESULTS: We have developed an information system mainly devoted to the storage and management of SNP genotype data produced by the Illumina platform from the raw outputs of genotyping into a relational database. The relational database can be accessed in order to import any existing data and export user-defined formats compatible with many different genetic analysis programs. After calculating family-based or case-control association study data, the results can be imported in SNPLims. One of the main features is to allow the user to rapidly identify and annotate statistically relevant polymorphisms from the large volume of data analyzed. Results can be easily visualized either graphically or creating ASCII comma separated format output files, which can be used as input to further analyses. CONCLUSIONS: The proposed infrastructure allows to manage a relatively large amount of genotypes for each sample and an arbitrary number of samples and phenotypes. Moreover, it enables the users to control the quality of the data and to perform the most common screening analyses and identify genes that become "candidate" for the disease under consideration. PMID- 18387202 TI - Is plant mitochondrial RNA editing a source of phylogenetic incongruence? An answer from in silico and in vivo data sets. AB - BACKGROUND: In plant mitochondria, the post-transcriptional RNA editing process converts C to U at a number of specific sites of the mRNA sequence and usually restores phylogenetically conserved codons and the encoded amino acid residues. Sites undergoing RNA editing evolve at a higher rate than sites not modified by the process. As a result, editing sites strongly affect the evolution of plant mitochondrial genomes, representing an important source of sequence variability and potentially informative characters. To date no clear and convincing evidence has established whether or not editing sites really affect the topology of reconstructed phylogenetic trees. For this reason, we investigated here the effect of RNA editing on the tree building process of twenty different plant mitochondrial gene sequences and by means of computer simulations. RESULTS: Based on our simulation study we suggest that the editing 'noise' in tree topology inference is mainly manifested at the cDNA level. In particular, editing sites tend to confuse tree topologies when artificial genomic and cDNA sequences are generated shorter than 500 bp and with an editing percentage higher than 5.0%. Similar results have been also obtained with genuine plant mitochondrial genes. In this latter instance, indeed, the topology incongruence increases when the editing percentage goes up from about 3.0 to 14.0%. However, when the average gene length is higher than 1,000 bp (rps3, matR and atp1) no differences in the comparison between inferred genomic and cDNA topologies could be detected. CONCLUSIONS: Our findings by the here reported in silico and in vivo computer simulation system seem to strongly suggest that editing sites contribute in the generation of misleading phylogenetic trees if the analyzed mitochondrial gene sequence is highly edited (higher than 3.0%) and reduced in length (shorter than 500 bp). In the current lack of direct experimental evidence the results presented here encourage, thus, the use of genomic mitochondrial rather than cDNA sequences for reconstructing phylogenetic events in land plants. PMID- 18387203 TI - SWS: accessing SRS sites contents through Web Services. AB - BACKGROUND: Web Services and Workflow Management Systems can support creation and deployment of network systems, able to automate data analysis and retrieval processes in biomedical research. Web Services have been implemented at bioinformatics centres and workflow systems have been proposed for biological data analysis. New databanks are often developed by taking into account these technologies, but many existing databases do not allow a programmatic access. Only a fraction of available databanks can thus be queried through programmatic interfaces. SRS is a well know indexing and search engine for biomedical databanks offering public access to many databanks and analysis tools. Unfortunately, these data are not easily and efficiently accessible through Web Services. RESULTS: We have developed 'SRS by WS' (SWS), a tool that makes information available in SRS sites accessible through Web Services. Information on known sites is maintained in a database, srsdb. SWS consists in a suite of WS that can query both srsdb, for information on sites and databases, and SRS sites. SWS returns results in a text-only format and can be accessed through a WSDL compliant client. SWS enables interoperability between workflow systems and SRS implementations, by also managing access to alternative sites, in order to cope with network and maintenance problems, and selecting the most up-to-date among available systems. CONCLUSIONS: Development and implementation of Web Services, allowing to make a programmatic access to an exhaustive set of biomedical databases can significantly improve automation of in-silico analysis. SWS supports this activity by making biological databanks that are managed in public SRS sites available through a programmatic interface. PMID- 18387204 TI - FunClust: a web server for the identification of structural motifs in a set of non-homologous protein structures. AB - BACKGROUND: The occurrence of very similar structural motifs brought about by different parts of non homologous proteins is often indicative of a common function. Indeed, relatively small local structures can mediate binding to a common partner, be it a protein, a nucleic acid, a cofactor or a substrate. While it is relatively easy to identify short amino acid or nucleotide sequence motifs in a given set of proteins or genes, and many methods do exist for this purpose, much more challenging is the identification of common local substructures, especially if they are formed by non consecutive residues in the sequence. RESULTS: Here we describe a publicly available tool, able to identify common structural motifs shared by different non homologous proteins in an unsupervised mode. The motifs can be as short as three residues and need not to be contiguous or even present in the same order in the sequence. Users can submit a set of protein structures deemed or not to share a common function (e.g. they bind similar ligands, or share a common epitope). The server finds and lists structural motifs composed of three or more spatially well conserved residues shared by at least three of the submitted structures. The method uses a local structural comparison algorithm to identify subsets of similar amino acids between each pair of input protein chains and a clustering procedure to group similarities shared among different structure pairs. CONCLUSIONS: FunClust is fast, completely sequence independent, and does not need an a priori knowledge of the motif to be found. The output consists of a list of aligned structural matches displayed in both tabular and graphical form. We show here examples of its usefulness by searching for the largest common structural motifs in test sets of non homologous proteins and showing that the identified motifs correspond to a known common functional feature. PMID- 18387205 TI - CellExcite: an efficient simulation environment for excitable cells. AB - BACKGROUND: Brain, heart and skeletal muscle share similar properties of excitable tissue, featuring both discrete behavior (all-or-nothing response to electrical activation) and continuous behavior (recovery to rest follows a temporal path, determined by multiple competing ion flows). Classical mathematical models of excitable cells involve complex systems of nonlinear differential equations. Such models not only impair formal analysis but also impose high computational demands on simulations, especially in large-scale 2-D and 3-D cell networks. In this paper, we show that by choosing Hybrid Automata as the modeling formalism, it is possible to construct a more abstract model of excitable cells that preserves the properties of interest while reducing the computational effort, thereby admitting the possibility of formal analysis and efficient simulation. RESULTS: We have developed CellExcite, a sophisticated simulation environment for excitable-cell networks. CellExcite allows the user to sketch a tissue of excitable cells, plan the stimuli to be applied during simulation, and customize the diffusion model. CellExcite adopts Hybrid Automata (HA) as the computational model in order to efficiently capture both discrete and continuous excitable-cell behavior. CONCLUSIONS: The CellExcite simulation framework for multicellular HA arrays exhibits significantly improved computational efficiency in large-scale simulations, thus opening the possibility for formal analysis based on HA theory. A demo of CellExcite is available at http://www.cs.sunysb.edu/~eha/. PMID- 18387206 TI - Discovering multi-level structures in bio-molecular data through the Bernstein inequality. AB - BACKGROUND: The unsupervised discovery of structures (i.e. clusterings) underlying data is a central issue in several branches of bioinformatics. Methods based on the concept of stability have been recently proposed to assess the reliability of a clustering procedure and to estimate the "optimal" number of clusters in bio-molecular data. A major problem with stability-based methods is the detection of multi-level structures (e.g. hierarchical functional classes of genes), and the assessment of their statistical significance. In this context, a chi-square based statistical test of hypothesis has been proposed; however, to assure the correctness of this technique some assumptions about the distribution of the data are needed. RESULTS: To assess the statistical significance and to discover multi-level structures in bio-molecular data, a new method based on Bernstein's inequality is proposed. This approach makes no assumptions about the distribution of the data, thus assuring a reliable application to a large range of bioinformatics problems. Results with synthetic and DNA microarray data show the effectiveness of the proposed method. CONCLUSIONS: The Bernstein test, due to its loose assumptions, is more sensitive than the chi-square test to the detection of multiple structures simultaneously present in the data. Nevertheless it is less selective, that is subject to more false positives, but adding independence assumptions, a more selective variant of the Bernstein inequality based test is also presented. The proposed methods can be applied to discover multiple structures and to assess their significance in different types of bio molecular data. PMID- 18387207 TI - Gene expression profile of peripheral blood mononuclear cells in response to HIV VLPs stimulation. AB - BACKGROUND: Baculovirus-expressed HIV-1 Pr55gag Virus-Like Particles (HIV-VLPs) induce maturation and activation of monocyte-derived dendritic cells (MDDCs) with a production of Th1- and Th2-specific cytokines. RESULTS: The analysis of genomic transcriptional profile of MDDCs, obtained from normal healthy donors and activated by HIV-VLPs, show the modulation of genes involved in the morphological and functional changes characterizing the MDDCs activation and maturation. Similar data are obtained using peripheral blood mononuclear cells (PBMCs), without further selection, showing the feasibility of a direct and "simplified" experimental procedure. CONCLUSIONS: The results here described show that the maturation pattern induced by HIV-VLPs in ex vivo generated MDDCs, can be observed also in CD14-expressing freshly derived PBMCs, with the possible identification of genetic predictors of individual response to immunogens. PMID- 18387208 TI - A three-state prediction of single point mutations on protein stability changes. AB - BACKGROUND: A basic question of protein structural studies is to which extent mutations affect the stability. This question may be addressed starting from sequence and/or from structure. In proteomics and genomics studies prediction of protein stability free energy change (DeltaDeltaG) upon single point mutation may also help the annotation process. The experimental DeltaDeltaG values are affected by uncertainty as measured by standard deviations. Most of the DeltaDeltaG values are nearly zero (about 32% of the DeltaDeltaG data set ranges from -0.5 to 0.5 kcal/mole) and both the value and sign of DeltaDeltaG may be either positive or negative for the same mutation blurring the relationship among mutations and expected DeltaDeltaG value. In order to overcome this problem we describe a new predictor that discriminates between 3 mutation classes: destabilizing mutations (DeltaDeltaG<-1.0 kcal/mol), stabilizing mutations (DeltaDeltaG>1.0 kcal/mole) and neutral mutations (-1.0or= 18 years) in the 2003 Medical Expenditure Panel Survey (MEPS). Given the idiosyncrasies of the cost data distribution, a recently developed extended estimating equation (EEE) model was employed to identify the relationship between patient satisfaction and healthcare expenditure, after controlling for individual demographic covariates, co-morbidity profile, and functional and activity limitations. A series of sensitivity analyses were conducted, in addition, to verify the identified relationship. All statistics were adjusted using the proper sampling weight from the MEPS data. RESULTS: Average annual healthcare expenditures for 2003 ranged between $3923 and $6073 when grouped by patient satisfaction ratings with a mean value $4779 for all individuals who rated perceived satisfaction of their healthcare. We found that there is no statistically significant relationship between patient satisfaction and total healthcare expenditure (p = 0.60) and a non-monotonic relationship is not identified either. All sensitivity analyses results revealed a lack of relationship between patient satisfaction and healthcare expenditures. LIMITATIONS: Patient satisfaction might not reflect the quality of healthcare from an objective clinical standpoint. The identified cost satisfaction relationship may not be extrapolated to other quality indicators. Due to the cross-sectional study design, no causal relationship could be inferred between patient satisfaction and healthcare expenditure. CONCLUSIONS: Our study adds to the literature on health care cost and quality by suggesting that the improvement of patient satisfaction may not require additional health care spending. PMID- 18387220 TI - New insights into the role of vitamin D and calcium in osteoporosis management: an expert roundtable discussion. AB - BACKGROUND: Adequate vitamin D and calcium nutrition play a critical role in the maintenance of musculoskeletal health and are considered the first step in osteoporosis treatment. ROUNDTABLE DISCUSSION: In February 2008 Merck Sharp & Dohme sponsored a 2-day, evidence-based expert panel on the benefits of vitamin D for the patient with osteoporosis and the role of vitamin D in combination with antiresorptive therapy for the management of osteoporosis. One of the primary objectives of the meeting was to review new data on the optimal serum 25-hydroxy vitamin D [25(OH)D] levels. The symposium was attended by 29 researchers and clinicians from Europe and the Middle East. The discussion focused on optimizing vitamin D and calcium nutrition and reducing falls and fractures in osteoporotic patients. CONCLUSIONS: Current evidence and expert opinion suggests that optimal serum 25(OH)D concentrations should be at least 50 nmol/L (20 ng/mL) in all individuals. This implies a population mean close to 75 nmol/L (30 ng/mL). In order to achieve this level, vitamin D intake of at least 20 microg daily is required. There is a wider therapeutic window for vitamin D than previously believed, and doses of 800 IU per day, regardless of sun exposure, season or additional multivitamin use, appear to present little risk of toxicity. Apart from fracture and fall prevention, optimization of vitamin D status may also have additional general health benefits. Based on newly emerging data regarding calcium supplementation, and recommendations for increased vitamin D intake, the current recommendations for calcium intake in postmenopausal women may be unnecessarily high. In addition to vitamin D and calcium, treatment of patients with osteoporosis at high risk of fractures should also include pharmacologic agents with proven vertebral and non-vertebral fracture efficacy. PMID- 18387221 TI - Appraisal of danazol prophylaxis for hereditary angioedema. AB - Hereditary angioedema (HAE) is an autosomal dominant disease characterized by painful, recurrent attacks of inflammation affecting the hands, feet, face, abdomen, urogenital tract, and the larynx. The inflammation can be disfiguring, debilitating, quite painful, and, in the case of laryngeal attacks, life threatening. Attacks are frequently the source of unnecessary exploratory abdominal procedures, extended hospital stays affecting a patient's ability to retain employment, and severe compromise of the patient's quality of life. HAE is estimated to affect 10,000 people in the US and is caused by deficient or dysfunctional C1-inhibitor, a naturally occurring molecule that is known to inhibit kallikrein, bradykinin, and other serine proteases in plasma. The treatment and management of HAE have been hampered by the dearth of safe and effective therapies. In the United States, there are currently no approved therapies for the treatment of acute HAE attacks. Although prophylactic HAE therapies do exist, they are often viewed as suboptimal due to moderate degrees of efficacy and the existence of adverse effects associated with therapy. Danazol, an attenuated androgen, is the most commonly prescribed prophylaxis treatment for HAE in the United States. Although it has demonstrated moderate efficacy in the prevention of HAE attacks, danazol's side-effect profile can be problematic because there is a correlation between frequency and severity of adverse events and dosage and duration of therapy. PMID- 18387222 TI - Efficacy of levocetirizine compared with montelukast in subjects with ragweed induced seasonal allergic rhinitis in the Environmental Exposure Unit. AB - Levocetirizine dihydrochloride, a potent H1-receptor antagonist, and montelukast sodium, a selective leukotriene receptor antagonist, have been approved for the treatment of seasonal allergic rhinitis (SAR), but target two different pathways that cause SAR symptoms. The study objective was to compare the efficacy of levocetirizine (LCTZ), 5 mg, and montelukast (MLKT), 10 mg, in reducing SAR symptoms in ragweed-sensitive adults exposed to ragweed pollen in the Environmental Exposure Unit (EEU). This randomized, double-blind, placebo controlled, parallel-group study of 418 adult subjects with SAR to ragweed compared the efficacy of LCTZ, MLKT, and placebo administered once daily (11:00 A.M.) for 2 consecutive days in the EEU. There were three evaluation periods: period I, 0-5 hours after first dose; period II, 22.5-24 hours after first dose; and period III, 0-4.5 hours after second dose. The primary efficacy variable was the Major Symptom Complex (MSC) score (six symptoms) over period I. Both active drugs significantly improved the MSC score compared with placebo in all periods. The adjusted mean MSC score difference between LCTZ and MLKT was -0.93 (p = 0.100) in period I, -3.11 (p < 0.001) in period II, -2.42 (p < 0.001) in period III, and -1.88 (p < 0.001) over the total treatment period. The same trends were observed for the Total Symptom Complex score (10 symptoms) and most individual symptoms. Subject-reported global satisfaction was greater for LCTZ compared with MLKT and placebo. All treatments had a favorable safety profile. LCTZ, 5 mg, was more effective than MLKT, 10 mg, in subjects with SAR and had better subject reported global satisfaction. PMID- 18387223 TI - April focus. PMID- 18387224 TI - Managing the acutely ill child. PMID- 18387225 TI - Place of death: how much does it matter? The priority is to improve end-of-life care in all settings. PMID- 18387226 TI - Collaboration in primary care: the need to see the bigger picture. PMID- 18387227 TI - The Tooke report: good news for general practice? PMID- 18387228 TI - Characteristics of children consulting for cough, sore throat, or earache. AB - BACKGROUND: GPs are often consulted for respiratory tract symptoms in children. AIM: To explore characteristics of children, their parents, and their GPs that are correlated with consulting a GP for cough, sore throat, or earache. DESIGN OF STUDY: Second Dutch National Survey of General Practice (DNSGP-2) with a health interview and an additional questionnaire. SETTING: Children aged 0-17 years registered with 122 GPs in Dutch general practice. METHOD: Characteristics of patients and their GPs were derived from the DNSGP-2 health interview and a questionnaire, respectively. Characteristics of the illness symptoms and GP consultation were acquired by means of an additional questionnaire. Data were analysed using multivariate logistic regression. RESULTS: Of all children who completed the questionnaire, 550 reported cough, sore throat, or earache in the 2 weeks preceding the interview with 147 of them consulting their GP. Young children more frequently consulted the GP for respiratory symptoms, as did children with fever, longer duration of symptoms, those reporting their health to be 'poor to good', and living in an urban area. When parents were worried, and when a child or their parents were cued by someone else, the GP was also consulted more often. GP-related determinants were not associated with GP consultation by children. CONCLUSION: This study emphasises the importance of establishing the reasons behind children with respiratory tract symptoms consulting their GP. When GPs are aware of possible determinants of the decision to consult a GP, more appropriate advice and reassurance can be given regarding these respiratory symptoms, which are generally self-limiting. PMID- 18387229 TI - Interprofessional relationships and communication in primary palliative care: impact of the Gold Standards Framework. AB - BACKGROUND: High quality end-of-life care in the community is achieved with effective multidisciplinary teamwork, interprofessional communication between GPs and district nurses, and early referral of patients to district nurses. These aspects of palliative care are highlighted in the Gold Standards Framework, a programme recently established in UK primary care. AIM: To investigate the extent to which the framework influences interprofessional relationships and communication, and to compare GPs' and nurses' experiences. DESIGN OF STUDY: Qualitative interview case study. SETTING: Fifteen participating practices from three primary care trusts in England. METHOD: Thirty-eight semi-structured interviews were undertaken with GPs, district nurses, Macmillan nurses, and framework facilitators. RESULTS: Adoption of the framework often resulted in earlier referral of palliative care patients to district nurses. Multidisciplinary team meetings enabled communication for sharing knowledge, discussing management problems, and keeping colleagues informed; however, arranging and maintaining such meetings was often problematic. Nurses particularly valued formal meetings while GPs generally preferred informal ad hoc dialogue. GPs largely maintained control of the mode of multidisciplinary working. The best functioning teams used a mixture of formal and informal meetings with a relatively non-hierarchical working style. CONCLUSION: Implementing the framework enabled processes of communication associated with high quality palliative care in general practice, but there was marked variation in how this worked in individual teams. In general, hierarchical doctor-nurse relationships persisted. PMID- 18387230 TI - The medical history as a diagnostic technology. AB - The medical history is a powerful diagnostic technology. However, in seeking to establish an appropriate balance between the history and the other diagnostic modalities more explicit consideration must be given to the performance characteristics of the medical history. Building on recent work undertaken in the UK and elsewhere in Europe it is now feasible to develop a library of setting specific likelihood ratios and kappa statistics for key elements of the medical history. Of particular importance to those working in primary care, statistically adjusted combinations of information from the medical history can be generated; furnishing clinicians with likelihood ratios of significant magnitudes. It is suggested that developing a more rational approach to the use of the medical history could lead to improvements in diagnostic efficiency and effectiveness, with benefits for individual patient care in addition to the overall NHS budget. When diagnosis is viewed as a processing pathway founded on a robust medical history, it becomes clear than in some situations investigations may become unnecessary and, in other circumstances, their impact will be enhanced. PMID- 18387231 TI - Minor surgery in general practice: are sterilised gloves necessary? PMID- 18387232 TI - GP's attitudes to hypertension guidelines. PMID- 18387233 TI - Centralised pathology services. PMID- 18387234 TI - Connecting for health. PMID- 18387236 TI - CAM. PMID- 18387237 TI - Limitations of the summary care record. PMID- 18387238 TI - Advertisements. PMID- 18387239 TI - Spinal manipulation. PMID- 18387240 TI - Management of headache. PMID- 18387241 TI - Appraisals. PMID- 18387243 TI - Antibiotic-associated diarrhoea. PMID- 18387244 TI - Sexual problems in later life. PMID- 18387245 TI - Headache management in general practice. PMID- 18387246 TI - Earwax. PMID- 18387247 TI - An alarm bell. PMID- 18387248 TI - Government strategy on alcohol: time to revise legislation and implement wider interventions. PMID- 18387249 TI - Real life ethics: autonomy versus duty of care. PMID- 18387250 TI - The Terracotta Army. PMID- 18387252 TI - Existentialism and the Mars bar diet. PMID- 18387251 TI - Climate change... what can we do? PMID- 18387253 TI - A patient's diary: episode 16 -- patient group meeting. PMID- 18387254 TI - Top tips in 2 minutes. PMID- 18387255 TI - Stigma. PMID- 18387256 TI - Dr. Grumpy will see you now... PMID- 18387257 TI - Selected topics in the field of spinal cord research. PMID- 18387258 TI - Stem and progenitor cell therapies: recent progress for spinal cord injury repair. AB - Mechanical trauma to the spinal cord is often accompanied by irreversible tissue damage, limited endogenous repair and permanent loss of motor, sensory and autonomic function. The implantation of exogenous cells or the stimulation of endogenous cells, to repopulate and replace or to provide a conducive environment for repair, offers a promising therapeutic direction for overcoming the multitude of obstacles facing successful recovery from spinal cord injury. Although relatively new to the scene of cell based therapies for reparative medicine, stem cells and their progenitors have been labeled as the 'cell of the future' for revolutionizing the treatment of CNS injury and neurodegenerative disorders. The following review examines the different types of stem cells and their progenitors, their utility in experimental models of spinal cord injury and explores the outstanding issues that still need to be addressed before they move towards clinical implementation. PMID- 18387259 TI - Adaptive changes in the injured spinal cord and their role in promoting functional recovery. AB - Although axons in the injured spinal cord are unable to regenerate, a modest recovery can often be found in both patients and animal models. One concept touted as a possible mechanism behind this recovery involves rearrangements in the brain and spinal cord, often referred to as plasticity. In the following review, centered around works from our laboratory, we will discuss plasticity throughout the entire central nervous system (CNS) induced by spinal cord injuries (SCIs) with an emphasis on sprouting of descending spinal tracts. Because this sprouting occurs spontaneously, it not only lends itself as a recovery mechanism, but also opens potential treatment avenues to promote further functional recovery. As such, various recent examples of approaches to pharmacologically promote plasticity within the spinal cord are discussed. PMID- 18387260 TI - Detection of the bladder volume from the neural afferent activities in dogs: experimental results. AB - OBJECTIVE: We evaluate the bladder volume and pressure through recording the bladder afferent activity in the sacral nerve roots in acute experiments of paraplegic dogs. These measurements are intended to report the status of the bladder and to adjust the stimulation parameters of an implantable electric stimulator. METHODS: The extraction of neural information for feedback in functional electrical stimulation is limited by the poor signal to noise ratio (SNR) in the sacral nerve recordings. We propose to inject a very low amplitude sinusoidal current with high SNR to the bladder through the nerve using a tripolar cuff electrode wrapped around the S2 nerve root. The application of this current (0.4 microA peak to peak, 30 Hz) allows detecting bladder afferent activity in its amplitude and the tissues impedance of the nerve. Acute experiments in dogs were performed to evaluate the proposed method. In each dog, the bladder infusion with saline was carried out at both slow and high filling rates. At the same time, the changes in the amplitude of the sinusoidal output voltage V(OUT) were recorded through the cuff nerve electrode. RESULTS: The data obtained from 26 acute experiments using eight dogs demonstrate that the amplitude of the recorded sinusoidal voltage V(OUT) varies proportionally with the bladder pressure during the bladder filling with saline solution. It also demonstrates that the bladder volume can be estimated from the increasing amplitude of the recorded V(OUT). CONCLUSION: This study shows that the increase in the V(OUT) is proportionally related to the increase in bladder pressure. The difference between the recorded V(OUT) during the bladder filling and the baseline V(OUT) can be a useful indicator of the changes in the bladder volume. PMID- 18387261 TI - Phase-dependent modulation of cutaneous reflexes in tibialis anterior muscle during passive stepping. AB - The purpose of this study was to determine whether the cutaneous reflex elicited in the tibialis anterior (TA) muscle would be modulated in a phase-dependent manner while human subjects were passively stepping on a treadmill (treadmill stepping) or in the air (air stepping). The passive stepping was produced by a robotic gait trainer, Lokomat. The cutaneous reflexes following electric stimulation to the distal tibial nerve were recorded at ten different phases of a step cycle under the condition of tonic dorsiflexion [10% of maximum electromyography activity (EMGmax)]. Cutaneous reflex EMG responses with peak latencies of 70-120 ms [middle latency responses (MLR)] were then analysed. The results showed that there were no visible differences in the background EMG activities at the ten phases or two passive stepping conditions. During treadmill stepping, however, the magnitude of the facilitatory reflex responses between the late stance and the early swing phase was strongly enhanced, whereas no clear modulation of the MLR during air stepping was observed. These results suggest that the load-related afferent information plays a key role in the modulation of the cutaneous reflex during human walking. PMID- 18387262 TI - Foot control in incomplete SCI: distinction between paresis and dexterity. AB - OBJECTIVE: To complement the clinical assessment of motor impairment after incomplete spinal cord injury (iSCI) by introducing a test that reliably distinguishes between muscle weakness (paresis) and impairment of dexterity in a simple foot motor task. METHODS: Auditory-paced ankle dorsi- and plantarflexion, in a supine position, was studied in 30 controls (to establish control values and to test reliability) and in 16 iSCI patients (test validation). The subjects were instructed to initiate dorsi- and plantarflexion as accurately in timing and with the largest range of motion (ROM) possible. For each frequency, accuracy of timing, ROM, peak velocity of dorsi- and plantarflexion and a time quotient for changing from dorsi- to plantarflexion and vice versa were determined. In iSCI subjects, these parameters were related to clinical measures of paresis, spasticity and proprioception. RESULTS: The test parameters showed good to very good reliability. The iSCI subjects were able to follow the target frequency with high accuracy, while ROM and peak velocity for dorsi- and plantarflexion were significantly reduced. Furthermore, there was a strong correlation between ROM/peak velocities and motor scores within the iSCI patients. DISCUSSION: Repetitive foot dorsi- and plantarflexion enables a distinction to be made between muscle weakness and reduced dexterity as the underlying cause of affected foot control. This distinction between and quantification of these two movement components complements the existing clinical examination, and in follow-up works, the recovery of these components may provide further insight into the mechanisms underlying motor function improvement after iSCI. PMID- 18387263 TI - Voices from the community--the voice of an oral cancer patient. PMID- 18387264 TI - Voices from the community--experiences from the dental office: initiating oral cancer screening. PMID- 18387265 TI - Why should dentists screen for oral cancer? PMID- 18387267 TI - Oral cancer: just the facts. AB - Oral cancer screening should be an integral part of a clinician's routine. This article reviews facts about oral cancer that are relevant to screening. The relevance of some issues in a particular dental practice will vary with the patient composition of the practice. PMID- 18387268 TI - Evaluation of a suspicious oral mucosal lesion. AB - Dentists who encounter a change in the oral mucosa of a patient must decide whether the abnormality requires further investigation. In this paper, we describe a systematic approach to the assessment of oral mucosal conditions that are thought likely to be premalignant or an early cancer. These steps, which include a comprehensive history, step-by-step clinical examination (including use of adjunctive visual tools), diagnostic testing and formulation of diagnosis, are routinely used in clinics affiliated with the British Columbia Oral Cancer Prevention Program (BC OCPP) and are recommended for consideration by dentists for use in daily practice. PMID- 18387266 TI - New hope for an oral cancer solution: together we can make a difference. AB - Oral cancer is associated with high mortality and morbidity rates, largely as a result of late diagnosis. Although dental practitioners are trained to identify premalignant and malignant lesions, an organized system is needed to offer guidance and to improve access to experts in diagnosis and management of these lesions. In this article, we describe the many ways in which the British Columbia Oral Cancer Prevention Program (BC OCPP) is addressing this challenge: by linking community dental practices and referral centres, by creating partnerships between scientists and clinicians that already have resulted in new technologies to enhance early diagnosis, by involving a broad range of stakeholders to ensure population-based screening and by engaging in provincial, national and international outreach. PMID- 18387269 TI - Biopsy and histopathologic diagnosis of oral premalignant and malignant lesions. AB - Accurate diagnosis of premalignant or malignant oral lesions depends on the quality of the biopsy, adequate clinical information and correct interpretation of the biopsy results. The purpose of this paper is to review the procedures for obtaining appropriate biopsy samples, and the criteria for diagnosing and grading dysplasias. The World Health Organization's description of the architectural and cytologic epithelial changes that characterize dysplasia is detailed, and guidelines for following up patients with premalignant and malignant lesions are provided. The benefits of using the centralized services and expertise of the British Columbia Oral Biopsy Service are also reviewed. PMID- 18387270 TI - [Nanotechnologies: from information sciences to pharmacology]. AB - The considerable breakthroughs of the physics during the last 30 years allowed us to conceive components, devices or materials at the nanoscale level and to manipulate them. Applications are already envisaged in the field of the medical and pharmaceutical sciences. These nanotechnologies will be applied to the biology as well as the diagnosis, the therapeutics and the functional rehabilitation (nanomedicine). Consequences in the pharmaceutical research and development are also possible in a near future. PMID- 18387271 TI - [Fifty years old and more patients' attitudes towards and experiences of generic substitution of prescription medicines]. AB - OBJECTIVE: Assess 50 years old and over patients' attitudes towards and experiences of generic substitution of prescription medicines. A special focus on information on patient attitude to generic drugs provided by their general practitioners (GPs). METHODS: Prospective study of patients in 15 general practices and in 2 retirements home was surveyed using a self-questionnaire. RESULTS: Four hundred and forty patients were included. Four twenty eleven per cent of the patients stated that they knew of the difference between brand-name drugs and generics but only 57% knew it exactly in fact. Seventy seven per cent had received generics by their GPs; 45% reported to have received information from their physician. The study found that patients who report to have received information from their physician about generic substitution were more likely to have switch. Patient in retirement home more frequently refused substitution. Elderly patients (75 and more) were wrong with generics definition compared with others patients, and observed more adverse effects after switching (20% versus 9% -p=0.027); patients made more mistake using generics than brand-name drugs (15.5% versus 7%--p<0.005); two thirds of the patients (72%) were satisfied with switching, and 57% reported to want more information and 85% of them that it comes from their GPs. CONCLUSION: Most of the patients are satisfied with generics. GPs are in an ideal position to inform their patients adequately about the equivalence of brand-name and generic drugs. Patient education is the best way to use generics in the future. More efforts must be devoted to provide adequate information to patients and GPs. PMID- 18387272 TI - [The need for a better definition of therapeutic indications of carboxypeptidase in delayed elimination of methotrexate]. AB - OBJECTIVES: High dose methotrexate (HD-MTX) may cause nonhaematological and haematological toxicities. MTX overexposure may be subsequent to administration errors or to delayed renal elimination resulting from MTX nephrotoxicity. The rescue agent carboxypeptidase rapidly hydrolyses MTX to inactive metabolites. However, current criteria for carboxypeptidase use are not well defined. We retrospectively evaluated the number of patients how should have received the drug if the criteria were applied, but who didn't receive it, and analysed their toxicities. METHODS: Patients treated at our institution in 2004 and 2005 and presenting the following criteria: concentrations of MTX at H48 >or= 3 microM, or impaired renal function, were considered. Post-course tolerance was recorded and graded. Recovery of renal function was followed-up periodically up to the 3rd month following the end of treatment. RESULTS: Twenty courses over 301 (7%) and 18 patients over 120 exhibited at least one criterion. Grade 3-4 toxicity was observed in 30% of the courses, including 2 severe acute renal impairment. Renal function decreased in most patients but had recovered upon the 3rd month in all but 2 patients. CONCLUSION: Despite a higher rate of toxicity than in general population, most of the patients recovered from it. Considering the cost of this treatment, the criteria for its therapeutic use could be restricted to concentration of MTX at H48 superior at 10 microM, associated with renal impairment. PMID- 18387273 TI - [Critical of the additive model of the randomized controlled trial]. AB - Randomized, double-blind, placebo-controlled clinical trials are currently the best way to demonstrate the clinical effectiveness of drugs. Its methodology relies on the method of difference (John Stuart Mill), through which the observed difference between two groups (drug vs placebo) can be attributed to the pharmacological effect of the drug being tested. However, this additive model can be questioned in the event of statistical interactions between the pharmacological and the placebo effects. Evidence in different domains has shown that the placebo effect can influence the effect of the active principle. This article evaluates the methodological, clinical and epistemological consequences of this phenomenon. Topics treated include extrapolating results, accounting for heterogeneous results, demonstrating the existence of several factors in the placebo effect, the necessity to take these factors into account for given symptoms or pathologies, as well as the problem of the "specific" effect. PMID- 18387274 TI - [Palifermin in the preventive treatment of oral mucositis in autograft patients: data on efficacy and tolerance following marketing approval]. AB - BACKGROUND: Few data are available on the efficacy of and tolerance to palifermin in the preventive treatment of oral mucositis in autograft patients treated with conditioning protocols usually used in France. METHODS: Our retrospective study bears on the first five candidates for autograft who benefited from prophylactic treatment with palifermin (60 microg/kg/day) between December 2005 and March 2006. RESULTS: Despite the prophylactic treatment, 3 patients developed severe oral mucositis. Moreover, in 3 patients, palifermin was found to be responsible for the onset of severe toxidermia, which required, in 2 cases, interruption of the treatment. CONCLUSION: These results need to be confirmed by a larger study that would also make it possible to evaluate the impact of palifermin on morbi mortality in autograft patients in France, and to determine the safety profile of this drug. PMID- 18387275 TI - [Systemic BCG reactions after intravesical BCG therapy. A report of four cases]. AB - Local Bacillus Calmette-Guerin (BCG) immunotherapy is an effective and widely used treatment for superficial bladder carcinoma. Local side effects are frequent, whereas systemic side effects are rare, but more serious. We report four cases of systemic BCG reaction. Although uncommon, this infectious complication of BCG therapy should always be considered in the appropriate clinical setting. The best approach to minimize this complication is a strict compliance with precautions and a close and rigorous surveillance of this drug. PMID- 18387276 TI - [Study of four cases of poisoning by Atractylis gummifera L. in Morocco]. AB - Atractylis gummifera L., a poisonous thistle responsible for frequent and sometimes mortal poisonings is known in Mediterranean region and in occident as this plant is used in alternative medicine. The clinical and biological data of four cases of poisoning in Morocco by Atractylis gummifera L. reported and discussed in this study will have to help to review this poisoning in order to inform better and to take care of the poisoned persons. Also this study may contribute to prevent the use of this plant in traditional medicine. PMID- 18387277 TI - [First meeting of the French CEIP (centres d'evaluation et d'information sur la pharmacodependance). Assessment of the abuse and pharmacodependence potential during drug development]. AB - The French system for the evaluation of abuse and dependence created in 1990 was definitely implemented in 1999 with the decree no99-249 making in particular mandatory the reporting of all serious cases of abuse or dependence to psychoactive drugs. This decree was also important to define the role of each party (regulatory agencies, pharmaceutical companies, health professionals and the network of the regional Centres for Evaluation and Information of Pharmacodependence) for all marketed psychoactive drugs in France. The first meeting on pharmacodependence was organized during the last annual congress of the French Society of Therapeutic Pharmacology and Physiology (P2T) held in Toulouse in April 2007. The aim of this meeting was that the role of the French system for the evaluation of abuse and dependence during the different steps of drug approval and after marketing in the context of real life would be better known. The French approach includes classical data obtained from experimental and clinical trials, but also and mainly data obtained from the specific tools installed since the creation of the CEIP. PMID- 18387279 TI - Interactions of Haemophilus parasuis and its LOS with porcine brain microvascular endothelial cells. AB - Haemophilus parasuis is a swine pathogen that causes Glasser's disease, which is characterized by polyserositis and meningitis. The pathogenesis of the H. parasuis infection is poorly understood. To cause meningitis, H. parasuis has to cross the blood-brain barrier (BBB) to gain access to the central nervous system (CNS). We recently showed that H. parasuis adheres to and invades porcine brain microvascular endothelial cells (PBMEC). The aim of this study was to evaluate the role of H. parasuis lipooligosaccharide (LOS) in the adhesion to PBMEC and to determine if H. parasuis (and/or its LOS) is able to induce apoptosis and activation of PBMEC. Results showed that adhesion of H. parasuis to PBMEC was partially mediated by LOS. Moreover, H. parasuis induces caspase-3-mediated apoptosis of PBMEC in a time--and dose--dependent manner, but its LOS did not seem to be involved in such a process. Furthermore, H. parasuis and, to a lesser extent, its LOS, was able to induce the release of IL-8 and IL-6 by PBMEC. Field strains of H. parasuis serotypes 4 and 5 induced similar levels of these inflammatory mediators. Our data suggest that H. parasuis uses cellular adhesion, induction of apoptosis and up-regulation of inflammatory mediators as mechanisms to invade the CNS via the BBB, and that LOS would play a certain but limited role in such pathological process. PMID- 18387281 TI - Modulation of the expression of the FGFR2-IIIb and FGFR2-IIIc molecules in dermatofibroma. PMID- 18387282 TI - Complete maternal isodisomy causing reduction to homozygosity for a novel LAMB3 mutation in Herlitz junctional epidermolysis bullosa. PMID- 18387283 TI - High 8-dehydrocholesterol level in a typical case of Conradi-Hunermann-Happle syndrome with a novel H76Y missense mutation. PMID- 18387284 TI - A hand-held apparatus for "nose-only" exposure of mice to inhalable microparticles as a dry powder inhalation targeting lung and airway macrophages. AB - Microparticles containing isoniazid and rifabutin were aerosolised using a simple apparatus fabricated from a 15-ml centrifuge tube. The dose available for inhalation by rodents was determined by collecting microparticles emitted at the delivery port. The dose available for inhalation was proportional to durations of exposure ranging from 10 to 90 s (10.5-13.5 CV%) and the weight of powder taken for fluidization (10-50 mg, r2=0.982). The apparatus was then used to administer inhalations of microparticles to mice. Other groups of mice received free rifabutin orally, or by i.v. injection. Rifabutin was estimated in serum and tissues of dosed mice by HPLC. When approximately 20 mg of microparticles were loaded in the apparatus, approximately 2.5 mg were collected at the delivery port in 30 s of operation. Mice inhaled approximately 300 microg of the 2.5 mg emitted at the delivery port. Airway and lung macrophages of mice receiving inhalations for 30 s accumulated 0.38 microg of rifabutin, while the amount in blood serum of these mice was 0.62 microg. In mice receiving 83 microg rifabutin i.v. or orally, the intracellular amounts were 0.06 and 0.07 microg respectively, while the amounts in serum were 1.02 and 0.80 microg. These observations confirmed that inhalation of microparticles targeted airway and lung macrophages. PMID- 18387285 TI - Comparison of killer toxin-producing and non-producing strains of Filobasidium capsuligenum: proposal for two varieties. AB - The basidiomycetous yeast, Filobasidium capsuligenum, produces killer toxin against the opportunistic pathogen Cryptococcus neoformans. Not every strain isolated so far is able to produce the anti cryptococcal toxin. The aim of the present work was to study the relationship between the toxins and the toxin producing and non-producing isolates. The toxin was coded on chromosomal DNA in each producing strain as molecular analysis revealed. In addition, both the killing spectra and biochemical properties of the toxins proved to be identical, thus intraspecific variation in the toxin was not found. For molecular typing of the isolates, the D1/D2 region of 26S rDNA, partial sequences of internal transcribed spacer (ITS) regions, PCR fingerprinting RAPD and mtDNA-RFLP patterns were examinated. Phylogenetic analyses based on the different approaches showed that strains with the ability of killer-toxin production and those without it differ significantly and cluster into two distinct groups. The differences between the two groups and the similarity within them suggest the authority to separate the species into varieties. PMID- 18387287 TI - Leukotriene D4 receptor antagonist montelukast alleviates water avoidance stress induced degeneration of the gastrointestinal mucosa. AB - We investigated the role of montelukast (ML), a cysteinyl leukotriene-1 receptor antagonist, on the water avoidance stress (WAS)-induced degeneration of the rat gastric, ileal and colonic mucosa. One group of Wistar albino rats were exposed to chronic WAS (WAS group) 2h daily for 5 days. Another group was administered ML (10mg/kg; i.p.; WAS+ML group) following every WAS exposure for 5 days. Control rats were injected with the vehicle solution only. The stomach, ileum and colon were dissected and investigated for histopathological changes with a light microscope as well as for topographical changes with a scanning electron microscope. The levels of malondialdehyde (MDA, a biomarker of oxidative damage) and glutathione (GSH, a biomarker of protective oxidative injury) were also determined in all dissected tissues. In the WAS group, the stomach epithelium showed ulceration in some areas, dilatations of the gastric glands, degeneration of gastric glandular cells, and prominent congestion of the capillaries. In a similar fashion, degenerated epithelium and severe vascular congestions were observed in the ileum and colon. In all the tissues dense inflammatory cell infiltration and mast cell degranulation in mucosa were observed. The levels of MDA were significantly increased whereas those of GSH were significantly decreased in all test tissues in the WAS group compared to the control group. The morphology of gastric, ileal and colonic mucosa in WAS+ML group showed a significant amelioration showing a reduction in inflammatory cell infiltration and mast cell degranulation. Increased MDA and decreased GSH levels in the WAS group were also ameliorated with ML treatment. Based on the results, ML supplement seems attenuated inflammatory effects of WAS induction in gastrointestinal mucosa. PMID- 18387286 TI - The regulation of aggrecanase ADAMTS-4 expression in human Achilles tendon and tendon-derived cells. AB - Several members of the ADAMTS (A Disintegrin And Metalloproteinase with ThromboSpondin motifs) family have been identified as aggrecanases, whose substrates include versican, the principal large proteoglycan in the tendon extracellular matrix. We have characterized the expression of ADAMTS-4 in human Achilles tendon and tendon-derived cells. ADAMTS-4 mRNA levels were higher in ruptured tendon compared with normal tendon or chronic painful tendinopathy. In tissue extracts probed by Western blotting, mature ADAMTS-4 (68 kDa) was detected only in ruptured tendons, while processed ADAMTS-4 (53 kDa) was detected also in chronic painful tendinopathy and in normal tendon. In cultured Achilles tendon cells, transforming growth factor-beta (TGF-beta) stimulated ADAMTS-4 mRNA expression (typically 20-fold after 24 h), while interleukin-1 induced a smaller, shorter-term stimulation which synergised markedly with that induced by TGF-beta. Increased levels of immunoreactive proteins consistent with mature and processed forms of ADAMTS-4 were detected in TGF-beta-stimulated cells. ADAMTS-4 mRNA was expressed at higher levels by tendon cells in collagen gels than in monolayer cultures. In contrast, the expression of ADAMTS-1 and -5 mRNA was lower in collagen gels compared with monolayers, and these mRNA showed smaller or opposite responses to growth factors and cytokines compared with that of ADAMTS-4 mRNA. We conclude that both ADAMTS-4 mRNA and ADAMTS-4 protein processing may be differentially regulated in normal and damaged tendons and that both the matrix environment and growth factors such as TGF-beta are potentially important factors controlling ADAMTS aggrecanase activities in tendon pathology. PMID- 18387288 TI - The impact of municipal solid waste treatment methods on greenhouse gas emissions in Lahore, Pakistan. AB - The contribution of existing municipal solid waste management to emission of greenhouse gases and the alternative scenarios to reduce emissions were analyzed for Data Ganj Bukhsh Town (DGBT) in Lahore, Pakistan using the life cycle assessment methodology. DGBT has a population of 1,624,169 people living in 232,024 dwellings. Total waste generated is 500,000 tons per year with an average per capita rate of 0.84kg per day. Alternative scenarios were developed and evaluated according to the environmental, economic, and social atmosphere of the study area. Solid waste management options considered include the collection and transportation of waste, collection of recyclables with single and mixed material bank container systems (SMBCS, MMBCS), material recovery facilities (MRF), composting, biogasification and landfilling. A life cycle inventory (LCI) of the six scenarios along with the baseline scenario was completed; this helped to quantify the CO2 equivalents, emitted and avoided, for energy consumption, production, fuel consumption, and methane (CH4) emissions. LCI results showed that the contribution of the baseline scenario to the global warming potential as CO2 equivalents was a maximum of 838,116 tons. The sixth scenario had a maximum reduction of GHG emissions in terms of CO2 equivalents of -33,773 tons, but the most workable scenario for the current situation in the study area is scenario 5. It saves 25% in CO2 equivalents compared to the baseline scenario. PMID- 18387289 TI - Size-fractionation and characterization of landfill leachate and the improvement of Cu2+ adsorption capacity in soil and aged refuse. AB - Leachate was collected from an anaerobic lagoon at Shanghai Laogang refuse landfill, the largest landfill in China, and the sample was separated into six fractions using micro-filtration membranes, followed by ultra-filtration membranes. Several parameters of the samples were measured, including chemical oxygen demand (COD), total organic carbon (TOC), total solids (TS), pH, total phosphate (TP), total nitrogen (TN), fixed solids (FS), NH4+, orthophosphate, color, turbidity, and conductivity. These parameters were then quantitatively correlated with the molecular weight cutoff of the membrane used. Organic matter in the dissolved fraction (MW<1kDa) predominated in the leachate, accounting for 65% of TOC. Thermal infrared spectroscopy was used to characterize the filter residues. Asymmetric and symmetric stretching of methyl and methylene groups, and of functional groups containing nitrogen and oxygen atoms, were observed. In addition, the ability of two different samples to adsorb heavy metals was tested. Cu2+ was chosen as the representative heavy metal in this study, and the samples were soil; aged refuse, which had spent 8 years in a conventional sanitary landfill; and samples of soil and aged refuse treated for 48h with leachate in the ratio of 5g of sample per 50ml of leachate. Cu2+ uptake by the raw soil was approximately 4.60microg/g, while uptake by the leachate-contacted soil and leachate-contacted aged refuse were 5.66 and 5.11microg/g, respectively. These results show that the organic matter in the leachate enhanced the capacity of aqueous solutions to adsorb Cu2+. PMID- 18387290 TI - Telomerase catalyzed fluorescent probes for sensitive protein profiling based on one-dimensional microfluidic beads array. AB - A nucleic acid-based signal amplified method for multiple proteins detection based on one-dimensional beads array using telomerase catalyzed fluorescent probes has been developed in this paper. The biotin labeled fluorescent probes were synthesized by telomerase in homogeneous solution. Approximately 360-480 fluorescein molecules were inserted in each probe. The limit of detection for p53 protein is 1.1 pM (S/N=3) and a 3 orders of linear dynamic range is obtained. The sensitivity is nearly two orders higher than the two-site "sandwich" immunoassay using the same platform. Using this method, cellular p53 protein contents of as few as 85 CNE2 cells per mul sample can be determined specifically. The expression changes of p53, c-myc and beta-actin in CNE2 cells were further examined as a function of anti-cancer drug treatment, and the results are consistent with our previous reports. Compared with immuno-polymerase chain reaction and immuno-rolling circle amplification, this method is simple, fast, cheap and suitable for multi-protein analysis. The multiplexed proteins profiling of cellular samples should facilitate the new opportunities to the fundamental research of tumor development and progression, especially to the low abundant tumor-associated proteins analysis. PMID- 18387291 TI - Amperometric glucose biosensor based on single-walled carbon nanohorns. AB - The biosensing application of single-walled carbon nanohorns (SWCNHs) was demonstrated through fabrication of an amperometric glucose biosensor. The biosensor was constructed by encapsulating glucose oxidase in the Nafion-SWCNHs composite film. The cyclic voltammograms for glucose oxidase immobilized on the composite film displayed a pair of well-defined and nearly symmetric redox peaks with a formal potential of -0.453 V. The biosensor had good electrocatalytic activity toward oxidation of glucose. To decrease detection potential, ferrocene monocarboxylic acid was used as a redox mediator. The mediated glucose biosensor shows a linear range from 0 to 6.0 mM. The biosensor shows high sensitivity (1.06 microA/mM) and stability, and can avoid the commonly coexisted interference. Because of impressive properties of SWCNHs, such as high purity and high surface area, SWCNHs and their composites are expected to be promising material for biomolecular immobilization and biosensing applications. PMID- 18387292 TI - Amperometric tyrosinase biosensor based on Fe3O4 nanoparticles-chitosan nanocomposite. AB - A novel tyrosinase biosensor based on Fe(3)O(4) nanoparticles-chitosan nanocomposite has been developed for the detection of phenolic compounds. The large surface area of Fe(3)O(4) nanoparticles and the porous morphology of chitosan led to a high loading of enzyme and the entrapped enzyme could retain its bioactivity. The tyrosinase-Fe(3)O(4) nanoparticle-chitosan bionanocomposite film was characterized with atomic force microscopy and AC impedance spectra. The prepared biosensor was used to determine phenolic compounds by amperometric detection of the biocatalytically liberated quinone at -0.2V vs. saturated calomel electrode (SCE). The different parameters, including working potential, pH of supporting electrolyte and temperature that governs the analytical performance of the biosensor have been studied in detail and optimized. The biosensor was applied to detect catechol with a linear range of 8.3 x 10(-8) to 7.0 x 10(-5)mol L(-1), and the detection limit of 2.5 x 10(-8)mol L(-1). The tyrosinase biosensor exhibits good repeatability and stability. Such new tyrosinase biosensor shows great promise for rapid, simple, and cost-effective analysis of phenolic contaminants in environmental samples. The proposed strategy can be extended for the development of other enzyme-based biosensors. PMID- 18387293 TI - Selective detection of uric acid in the presence of ascorbic acid at physiological pH by using a beta-cyclodextrin modified copolymer of sulfanilic acid and N-acetylaniline. AB - A beta-cyclodextrin (CD) modified copolymer membrane of sulfanilic acid (p-ASA) and N-acetylaniline (SPNAANI) on glassy carbon electrode (GCE) was prepared and used to determine uric acid (UA) in the presence of a large excess of ascorbic acid (AA) by differential pulse voltammetry (DPV). The properties of the copolymer were characterized by X-ray photoelectron spectra (XPS) and Raman spectroscopy. The oxidation peaks of AA and UA were well separated at the composite membrane modified electrode in phosphate buffer solution (PBS, pH 7.4). A linear relationship between the peak current and the concentration of UA was obtained in the range from 1.0 x 10(-5) to 3.5 x 10(-4)mol L(-1), and the detection limit was 2.7 x 10(-6)mol L(-1) at a signal-to-noise ratio of 3. Two hundred and fifty-fold excess of AA did not interfere with the determination of UA. The application of the prepared electrode was demonstrated by measuring UA in human serum samples without any pretreatment, and the results were comparatively in agreement with the spectrometric clinical assay method. PMID- 18387294 TI - Towards a systems level analysis of health and nutrition. AB - Although theoretical systems analysis has been available for over half a century, the recent advent of omic high-throughput analytical platforms along with the integration of individual tools and technologies has given rise to the field of modern systems biology. Coupled with information technology, bioinformatics, knowledge management and powerful mathematical models, systems biology has opened up new vistas in our understanding of complex biological systems. Currently there are two distinct approaches that include the inductively driven computational systems biology (bottom-up approach) and the deductive data-driven top-down analysis. Such approaches offer enormous potential in the elucidation of disease as well as defining key pathways and networks involved in optimal human health and nutrition. The tools and technologies now available in systems biology analyses offer exciting opportunities to develop the emerging areas of personalized medicine and individual nutritional profiling. PMID- 18387295 TI - Nutrigenomics research for personalized nutrition and medicine. AB - Current nutritional and genetic epidemiological methods yield 'risk factors' on the basis of population studies. Risk factors, however, are statistical estimates of the percentage reduction in disease in the population if the risk were to be avoided or the gene variant were not present. These measures are often assumed to apply to individuals who are likely to differ in genetic make-up, lifestyle, and dietary patterns than to the individuals in the study population. Developing individual risk factors in light of the genetic diversity of human populations, the complexity of foods, culture and lifestyle, and the variety of metabolic processes that lead to health or disease is a significant challenge for personalizing dietary advice for healthy or individuals with chronic disease. PMID- 18387296 TI - Systematic review of the association between circulating interleukin-6 (IL-6) and cancer. AB - Our aim was to systematically review the epidemiologic evidence for an association of circulating interleukin-6 (IL-6), an inflammatory cytokine and cancer. We systematically searched electronic databases Embase, Medline and Web of Science for the studies of circulating IL-6 and any form of cancer. We identified and reviewed 189 discrete studies, consisting of 177 prevalent studies and three prospective studies. Cancer patients' IL-6 concentrations were higher than healthy controls' in most studies, but the results of investigations comparing IL-6 in cancer patients and individuals with benign diseases were less consistent. Due to the small number of prospective studies it is impossible to determine whether IL-6 is causally related to cancer. Large prospective studies of circulating IL-6 or studies using the functional variants of the IL-6 gene as instruments for circulating IL-6 concentrations would provide information on possible aetiological links between IL-6 and malignancy. PMID- 18387297 TI - Nitrate removal from groundwater using constructed wetlands under various hydraulic loading rates. AB - This study set up two flow-through pilot-scale constructed wetlands with the same size but various flow patterns (free water surface flow (FWS) and subsurface flow (SSF)) to receive a nitrate-contaminated groundwater. The effects of hydraulic loading rate (HLR) on nitrate removal as well as the difference in performance between the various types of wetlands were investigated. Nitrate removal rates of both wetlands increased with increasing HLR until a maximum value was reached. The maximum removal rates, occurred at HLR of 0.12 and 0.07 m d(-1), were 0.910 and 1.161 g N m(-2)d(-1) for the FWS and SSF wetland, respectively. After the maximum values were reached, further increasing HLR led to a considerable decrease in nitrate removal rate. Nitrate removal efficiencies remained high (>85%) and effluent nitrate concentrations always satisfied drinking water standard (<10mg NO3-NL(-1)) when HLR did not exceed 0.04 m d(-1) for both FWS and SSF wetlands. The first-order nitrate removal rate constant tends to decrease with increasing HLRs. The FWS wetland provided significantly higher (p<0.05) organic carbon in effluent than the SSF wetland, while the SSF wetland exhibited significantly (p<0.05) lower effluent DO than the FWS wetland. However, there was no significant difference (p>0.05) in nitrate removal performance between the two types of constructed wetlands in this study except in one trial operating at HLR of 0.06-0.07 m d(-1). PMID- 18387298 TI - Removal of free fatty acid in waste frying oil by esterification with methanol on zeolite catalysts. AB - The removal of free fatty acid (FFA) in waste frying oil by esterification with methanol was conducted using various zeolite catalysts. The ZSM-5 (MFI), mordenite (MOR), faujasite (FAU), beta (BEA) zeolites, and silicalite were employed with different Si/Al molar ratio in the reaction. The effects of acidic properties and pore structure of the zeolite catalysts were discussed relating to the conversion of the FFA. The MFI zeolite induced an improvement of the removal efficiency of FFA by cracking to the FFA in its pore structure due to its narrow pore mouth. The catalytic activity for FFA removal was lowered with decreasing of acid strength of the zeolites. The strong acid sites of zeolites induced the high conversion of FFA comparatively. The acid strength and pore structure of acidic zeolites affected the catalytic activity in FFA removal. PMID- 18387299 TI - Comparative study on application of T.lanuginosus SSBP xylanase and commercial xylanase on biobleaching of non wood pulps. AB - Biobleaching of three non wood kraft pulps (bagasse, rice straw and wheat straw) by Thermomyces lanuginosus SSBP xylanase and commercial xylanase (cartazyme sandoz), was studied in order to investigate their potential and effect on their various properties (reduction sugars, chlorine dioxide, kappa number, brightness and chromophores). In generally, xylanases released chromophores and reducing sugars and decreased kappa number of pulps. These samples gained over six brightness points over controls. Biobleaching of rice straw pulp with xylanase cartazyme (Sandoz) produced chlorine dioxide savings of up to 25% or 3.5-4 kg chlorine dioxide/ton pulp. PMID- 18387300 TI - Design and synthesis of morpholine derivatives. SAR for dual serotonin & noradrenaline reuptake inhibition. AB - Single enantiomer (SS) and (RR) 2-[(phenoxy)(phenyl)methyl]morpholine derivatives 5, 8-23 are inhibitors of monoamine reuptake. Target compounds were prepared using an enantioselective synthesis employing a highly specific enzyme-catalysed resolution of racemic n-butyl 4-benzylmorpholine-2-carboxylate (26) as the key step. Structure-activity relationships established that serotonin and noradrenaline reuptake inhibition are functions of stereochemistry and aryl/aryloxy ring substitution. Consequently, selective SRI, selective NRI and dual SNRIs were all identified. One of these compounds, a potent and selective dual SNRI, (SS)-5a was selected as a candidate for further pre-clinical evaluation. PMID- 18387301 TI - From hope and expectation to unexpected death after cardiac surgery. AB - OBJECTIVE: Relatives of patients undergoing cardiac surgery expect successful outcome but sometimes complications cause death. The aim was to interview relatives of patients who have died in connection with cardiac surgery and describe their experiences of information, reception and care. METHODS: Data were obtained from semi-structured interviews with 18 relatives of deceased patients and then analysed using qualitative content analysis. RESULTS: Two main groups emerged: "Analysing the situation" with the sub-groups: knowledge of cardiac disease, the road to operation, hope and despair, information and choice and "The thin thread of life" with the sub-groups, reception, life is over, care, death as a relief, cause of death and support. CONCLUSION: Most relatives were satisfied with the information and care in connection with the operation and at the end of life. However, some aspects such as inadequate pain control and transportation of critically ill patients to other wards and hospitals could be improved. One way is to introduce a co-ordinator in order to better support patients, next of kin and colleagues without experience of cardio-thoracic surgery who need help during the patients' way from diagnosis and acceptance of cardiac surgery and through the treatment and postoperative care. PMID- 18387302 TI - Isolated chorda tympani injury following petrous temporal fracture. AB - Petrous temporal fractures account for nearly a fifth of all skull base fractures but are associated with taste disturbance in only 2% of cases. Chorda tympani injuries almost invariably occur in association with other nerve injuries. We present a rare case of an isolated chorda tympani injury associated with a petrous temporal fracture. Our observations suggest that the gustatory fibres in this nerve are segregated into groups subserving different taste modalities. PMID- 18387303 TI - Aetiology and prognosis of encephalopathic patterns on electroencephalogram in a general hospital. AB - The purpose of this study was to investigate the frequency and clinical outcome of patients with encephalopathic electroencephalograms (EEGs) in a neurophysiology department based in a general hospital. We performed a retrospective review of all EEGs obtained during an 18-month period in a large tertiary referral hospital. The referral reasons for EEG, the diagnoses reached, and patient outcomes were reviewed according to EEG severity. One hundred and twenty-three patients with encephalopathic EEGs were reviewed. The most common referral reason found was for an assessment of a possible first-onset seizure. The most common diagnosis found was one of dementia or learning disability. Of patients who were followed-up for a median of 19 months, 20.7% had died. The mortality rate generally increased according to the severity of the encephalopathy on EEG. However, 21.4% of those patients with excessive theta activity only on EEG had died. This study highlights an increased mortality even in the apparently 'milder' degrees of EEG abnormalities. PMID- 18387304 TI - Structure-based discovery of a novel non-peptidic small molecular inhibitor of caspase-3. AB - Ac-DNLD-CHO is a novel caspase-3 specific peptide inhibitor that was rationally designed by our computational strategy. The specificity was shown to be due to the specific interaction of NLD moiety with the active site of caspase-3 on the basis of docking mode and site-directed mutagenesis analyses. Here, we computationally screened non-peptidic small molecular inhibitors of caspase-3 from our chemical library using a reliable pharmacophore derived from the specific binding mode of NLD. Through in vitro enzyme assay of the screened candidate compounds, we discovered a novel caspase-3 specific small molecular inhibitor, CS4566, which has a unique scaffold structure. The binding mode of CS4566 to caspase-3 mimics that of NLD, especially LD moiety. This represents a promising lead compound for creating non-peptidic pharmaceuticals for caspase mediated diseases, such as neurodegenerative disorders. PMID- 18387305 TI - Cross-section measurement for the 10B(n, alpha)7Li reaction at 4.0 and 5.0 MeV. AB - Cross-sections of the (10)B(n, alpha)(7)Li reaction were measured at En=4.0 and 5.0 MeV. A gridded ionization chamber (GIC) was used as charged particle detector. Neutrons were produced through the D(d, n)(3)He reaction with a deuterium gas target. Experiments were performed at the 4.5 MV Van de Graaff accelerator of Peking University. Cross-section data of the (238)U(n, f) reaction were employed as standard. The measured cross-sections of the (10)B(n, alpha)(7)Li reaction at 4.0 and 5.0 MeV are 211+/-17 and 169+/-14 mb, respectively, and they are compared with existing results of measurements and evaluations. PMID- 18387306 TI - Recurrent massive aneurysm of the ascending aorta after aortic root replacement with inclusion cylinder technique. PMID- 18387307 TI - Extra-thoracic hematoma after minor blunt chest injury. PMID- 18387308 TI - The F-techniques: advances in receptor protein studies. AB - Recent developments in advanced microscopy techniques, the so-called F techniques, including Forster resonance energy transfer, fluorescence correlation spectroscopy and fluorescence lifetime imaging, have led to a wide range of novel applications in biology. The F-techniques provide quantitative information on biomolecules and their interactions and give high spatial and temporal resolution. In particular, their application to receptor protein studies has led to new insights into receptor localization, oligomerization, activation and function in vivo. This review focuses on the application of the F-techniques to the study of receptor molecules and mechanisms in the last three years and provides information on new modalities that will further improve their applicability and widen the range of biological questions that can be addressed. PMID- 18387309 TI - Cytocidal effect of interleukin 1 (IL-1) on HeLa cells is mediated by both soluble and transmembrane tumor necrosis factor (TNF). AB - Interleukin 1 (IL-1) is a pleiotropic cytokine able to induce cytocidal effect. The aim of the presented work was to analyze the mechanism of IL-1-induced cytocidal effect in HeLa cells in the presence of cycloheximide (CHX). We found that the pattern of IL-1-induced cell death shares significant similarities with the effect of tumor necrosis factor (TNF) in these cells. Subsequently, we identified IL-1 cytotoxicity as an indirect effect. The supernatant collected from the cells treated with IL-1 and CHX showed toxic activity towards IL-1 resistant while TNF-sensitive A9 cells. Furthermore, antibodies neutralizing TNF blocked HeLa cell death induced by IL-1/CHX. TNF was then detected in HeLa cells by means of flow cytometry, fluorescence microscopy and ELISA of detergent soluble cell extracts. In the presence of an inhibitor of TNF sheddase (TACE), the cytotoxic effect of IL-1/CHX and the amount of TNF protein in detergent soluble cell extracts were enhanced. These results suggest that in response to interleukin 1/CHX, the amount of transmembrane TNF is increased. Taken together, we demonstrated that the mechanism of IL-1 cytotoxic activity in HeLa cells in the presence of CHX depends on the function of soluble and transmembrane TNF. PMID- 18387310 TI - Endogenous IL-18 in experimentally induced asthma affects cytokine serum levels but is irrelevant for clinical symptoms. AB - T cells and T cell derived cytokines are involved in the complex pathogenesis of asthma. The role of the cytokine IL-18 however, is not clearly defined so far. On the one hand side IL-18 induces Th1-type cytokines and thereby might counter regulate Th2-mediated allergic asthma. On the other hand IL-18 also bears pro inflammatory effects possibly enhancing experimental asthma. In order to elucidate the role of IL-18 in allergic pulmonary inflammation typical symptoms were compared after induction of experimental asthma in IL-18(-/-) and in wild type mice. Asthma was induced using ovalbumin (OVA) as allergen for sensitization and challenge. Sham sensitized and OVA challenged mice served as controls. Bronchoalveolar lavage-fluid cytology, leukocyte infiltration in lung tissues, serum levels of OVA-specific IgE and cytokines, and lung function were analyzed. Clear differences could be observed between control and asthmatic mice, both in wild type and IL-18(-/-) animals. Surprisingly, no differences were found between asthmatic wild type and IL-18(-/-) mice. Thus, in contrast to conflicting data in the literature IL-18 did not suppress or enhance the pulmonary allergic immune response in a murine experimental model of asthma. PMID- 18387311 TI - Charge assisted laser desorption/ionization mass spectrometry of droplets. AB - We propose and evaluate a new mechanism to account for analyte ion signal enhancement in ultraviolet-laser desorption mass spectrometry of droplets in the presence of corona ions. Our new insights are based on timing control of corona ion production, laser desorption, and peptide ion extraction achieved by a novel pulsed corona apparatus. We demonstrate that droplet charging rather than gas phase ion-neutral reactions is the major contributor to analyte ion generation from an electrically isolated droplet. Implications of the new mechanism, termed charge assisted laser desorption/ionization (CALDI), are discussed and contrasted with those of the laser desorption atmospheric pressure chemical ionization method (LD-APCI). It is also demonstrated that analyte ion generation in CALDI occurs with external electric fields about one order of magnitude lower than those needed for atmospheric pressure matrix assisted laser desorption/ionization or electrospray ionization of droplets. PMID- 18387312 TI - Recombinant human fibrinogen expressed in the yeast Pichia pastoris was assembled and biologically active. AB - Fibrinogen is a large plasma glycoprotein with a molecular mass of 340kDa that plays a critical role in the final stage of blood coagulation. Human plasma fibrinogen is a dimeric molecule comprising two sets of three different polypeptides (Aalpha, 66kDa; Bbeta, 55kDa; gamma, 48kDa). To express recombinant human fibrinogen in the methylotrophic yeast Pichia pastoris, we constructed an expression vector containing three individual fibrinogen chain cDNAs under the control of the mutated AOX2 (mAOX2) promoter. First, P. pastoris GTS115 was transformed with the vector, but the expressed recombinant fibrinogen suffered severe degradation by yeast-derived proteases under conventional nutrient culture conditions. Fibrinogen degradation was prevented by using the protease A deficient strain SMD1168 as a host strain and regulating the pH of the culture to between 5.5 and 7.0. Western blot analysis revealed that the Aalpha, Bbeta and gamma chains of recombinant fibrinogen were assembled and secreted as a complete molecule. The Bbeta chain of the recombinant fibrinogen was N-glycosylated but the Aalpha chain, as in plasma fibrinogen, was not. The gamma chains however were heterologous, one being N-glycosylated and the other not. The recombinant fibrinogen was capable of forming a thrombin-induced clot in the presence of factor XIIIa and both the glycosylated and the non-glycosylated gamma chains were involved in the formation of cross-linking fibrin. The present study indicates that the recombinant fibrinogen expressed in P. pastoris, although different from plasma fibrinogen in post-translational modification, is correctly assembled and biologically active. PMID- 18387314 TI - Innervation zone shift at different levels of isometric contraction in the biceps brachii muscle. AB - Experiments were carried out to examine whether innervation zone (IZ) location remains stable at different levels of isometric contraction in the biceps brachii muscle (BB), and to determine how the proximity of the IZ affects common surface electromyography (sEMG) parameters. Twelve subjects performed maximal (MVC) and submaximal voluntary isometric contractions at 10%, 20%, 30%, 40%, 50% and 75% of MVC. sEMG signals were recorded with a 13 rows x 5 columns grid of electrodes from the short head of BB. The IZ shifted in the proximal direction by up to 2.4 cm, depending upon the subject and electrode column. The mean shift of all the columns was 0.6+/-0.4 cm (10% vs. 100% MVC, P<0.001). This shift biased the average values of mean frequency (+21.8+/-9.9 Hz, P<0.001), root mean square ( 0.16+/-0.15 mV, P<0.05) and conduction velocity (-1.15+/-0.93 m/s, P<0.01) in the channels immediately proximal to the IZ. The shift in IZ could be explained by shortening of the muscle fibers, and thus lengthening of the (distal) tendon due to increasing force. These results underline the importance of individual investigation of IZ locations before the placement of sEMG electrodes, even in isometric contractions. PMID- 18387313 TI - Quantitative 3-D imaging of eukaryotic cells using soft X-ray tomography. AB - Imaging has long been one of the principal techniques used in biological and biomedical research. Indeed, the field of cell biology grew out of the first electron microscopy images of organelles in a cell. Since this landmark event, much work has been carried out to image and classify the organelles in eukaryotic cells using electron microscopy. Fluorescently labeled organelles can now be tracked in live cells, and recently, powerful light microscope techniques have pushed the limit of optical resolution to image single molecules. In this paper, we describe the use of soft X-ray tomography, a new tool for quantitative imaging of organelle structure and distribution in whole, fully hydrated eukaryotic Schizosaccharomyces pombe cells. In addition to imaging intact cells, soft X-ray tomography has the advantage of not requiring the use of any staining or fixation protocols--cells are simply transferred from their growth environment to a sample holder and immediately cryofixed. In this way the cells can be imaged in a near native state. Soft X-ray tomography is also capable of imaging relatively large numbers of cells in a short period of time, and is therefore a technique that has the potential to produce information on organelle morphology from statistically significant numbers of cells. PMID- 18387315 TI - Reticulate evolution in the natural range of the invasive wetland tree species Melaleuca quinquenervia. AB - The Melaleuca leucadendra complex (broad-leaf paperbarks; Myrtaceae) is a dominant component of the tropical and sub-tropical biota of Australia, particularly in wetlands of high conservation significance. In Florida and other parts of the Americas, however, one member of the group (Melaleuca quinquenervia) is a serious ecological and economic weed. Understanding the relationships and evolution of the group is integral to both conservation and biocontrol efforts. Although the complex is currently considered to include up to 14 species, there has been some concern over taxonomic boundaries within the complex because most species are circumscribed only by combinations of characters, each of which also occurs in other species. Here, DNA sequence data derived from the chloroplast and two nuclear regions are used to explore the relationships of M. quinquenervia. We find little evidence for clear species boundaries within the M. leucadendra complex in general, with regional sharing of chloroplast haplotypes across morphologically defined taxa, indicating asymmetrical introgression or retention of ancestral haplotypes (lineage sorting). Phylogenies were further confounded by the recovery of multiple copies of both nuclear regions sequenced (ITS and rpb2) from many individuals. There was no clear evidence of polyploidy or pseudogenes, but multiple duplications of rpb2 could not be ruled out. Parsimony networks of the nuclear ITS region show some clustering of haplotypes by morphospecies but there is also evidence of both hybridisation and recombination. Signals of introgression were also evident in rpb2, supporting an hypothesis of recent or ongoing gene flow between M. quinquenervia and other members of the M. leucadendra complex. Both relaxed and fixed molecular-clock dating estimate the introgression to have occurred sometime within the past seven million years (95% CI: 0.7-18). The New Caledonian population of M. quinquenervia appears to have been established by dispersal from Australia during this period. M. quinquenervia is found to have alleles closely related to multiple different morphotaxa within the M. leucadendra complex, suggesting considerable past introgression into this taxon from some other members of the M. leucadendra complex, and this has implications for biocontrol efforts. The M. leucadendra complex appears to reflect early to intermediate stages of speciation, possibly driven by different ecologies. PMID- 18387316 TI - Total shoulder arthroplasty using the superior approach: influence on glenoid loosening and superior migration in the long-term follow-up after Neer II prosthesis installation. AB - Glenoid component loosening and superior humeral translation are common after Neer II total shoulder arthroplasty using the anterior approach. To determine whether the superior approach reduced these complications, we retrospectively reviewed 20 shoulders in 16 patients. Both components were cemented. Patient satisfaction, unweighted Constant score, and imaging studies were evaluated at a mean of 3.5 years and at a mean of 11.1 years. Fourteen patients were satisfied or very satisfied. The mean unweighted Constant score improved from 25/100 preoperatively to 57/100 after 3.5 years and to 51/100 after 11.1 years. Pain relief contrasted with low strength. Radiolucent lines appeared around 95% of glenoid components and 20% of humeral stems. Computed tomography showed severe glenoid osteolysis in 3 of 13 shoulders. Humeral superior translation did not occur. This study confirms the glenoid component fixation issue. The superior approach may reduce the risk of humeral superior translation and radiologic glenoid component loosening. PMID- 18387317 TI - Shoulder joint position sense after thermal, open, and arthroscopic capsulorrhaphy for recurrent anterior instability. AB - Identifying the optimal surgical treatment for recurrent, anterior glenohumeral instability remains a challenge. Our purpose was to compare shoulder joint position sense among open, arthroscopic, and thermal capsulorrhaphy patients after repair of recurrent anterior instability and to compare these patients to healthy, control subjects. Sixty-seven adults (45 post-surgical patients, 22 controls) volunteered to participate in the study. We evaluated both the surgically repaired and contralateral shoulders of 45 capsulorrhaphy patients (28 men, 17 women) and compared their results with the normal bilateral shoulders of 22 age-matched controls (11 men, 11 women). Accuracy of joint position sense was quantified via passive reproduction of target positions set at 60% and 90% of each subject's maximum passive external rotation (ER(max)). We observed no significant differences in joint position sense between the repaired shoulders and the contralateral normal shoulders of all groups of capsulorrhaphy patients. Open and thermal capsulorrhaphy patients demonstrated significantly better (P or = 60 degrees C), the M0 value of bitumen at lower temperatures (<60 degrees C) can be corrected by Curie's Law. Consequently, some important petrophysical properties of bitumen, such as hydrogen index (HI), fluid content and viscosity were evaluated by using corrected T2. PMID- 18387324 TI - Role of adenomatous polyposis coli (APC) and microtubules in directional cell migration and neuronal polarization. AB - In response to extracellular signals during embryonic development, cells undergo directional movements to specific sites and establish proper connections to other cells to form organs and tissues. Cell extension and migration in the direction of extracellular cues is mediated by the actin and microtubule cytoskeletons, and recent results have shed new light on how these pathways are activated by neurotrophins, Wnt or extracellular matrix. These signals lead to modifications of microtubule-associated proteins (MAPs) and point to glycogen synthase kinase (GSK) 3beta as a key regulator of microtubule function during directional migration. This review will summarize these results and then focus on the role of microtubule-binding protein adenomatous polyposis coli (APC) in neuronal polarization and directed migration, and on its regulation by GSK3beta. PMID- 18387326 TI - Screening of persistent organic pollutants by QSPR classification models: a comparative study. AB - A Quantitative Structure-Property Relationships (QSPRs) study for the prediction of the environmental persistence of a set of 250 heterogeneous organic compounds is here presented. Three a priori defined classes of environmental persistence were generated, by Hierarchical Cluster Analysis, from the combination of half life data in air, water, soil and sediment available for all the studied compounds. QSPR classification models were successfully developed using different techniques (k-NN, CART and CP-ANN) and three interpretable theoretical molecular descriptors. Robust external validation was provided by statistical splitting and also on completely new data. The good performances of all these models were compared and their structural domains were analyzed. The analysis of the errors highlights a slight tendency of persistence overestimation, misclassifying chemicals from a lower to a higher class of persistence, in line with the precautionary principle. Finally, the reliability of the proposed QSPR models was verified further with new data from the literature. The structure-based classification models, applicable for the prediction of potential persistence of heterogeneous organic compounds, could be useful as preliminary support tools for the identification and prioritization of new potential POPs among already existing chemicals as well as "screening prior to synthesis" procedures to avoid the production, and consequent release into the environment, of new POPs. PMID- 18387327 TI - The role of thyroid hormones during the development of eye pigmentation in the Pacific bluefin tuna (Thunnus orientalis). AB - Thyroid hormones (THs) are essential for the embryonic and post-embryonic development of fish. We studied the role of THs during the early, post-embryonic, development of Pacific bluefin tuna. Embryos were treated with L-thyroxine (T(4)) or the anti-thyroid drug methimazole (MMI), and reared in microtitre plates for 3 days. Immersion in MMI, but not T(4), led to retardation of retinal pigment epithelium (RPE) pigmentation 3 days post-hatching (dph). Concurrent immersion in T(4) and MMI had no effect of RPE pigmentation. We also measured the expression of TRalphaA, TRalphaB, and TRbeta mRNA using real-time RT-PCR. Treatment with MMI significantly reduced TRbeta mRNA expression. Taken together these results suggest that the development of RPE pigmentation is mediated by TH, most likely via TRbeta. PMID- 18387328 TI - Beneficial effects of substrate reduction therapy in a mouse model of GM1 gangliosidosis. AB - GM1 gangliosidosis is an inherited neurodegenerative disorder caused by lysosomal beta-galactosidase deficiency, resulting in the storage of GM1 and GA1, primarily in the central nervous system. This disease typically afflicts infants and young children and there is currently no effective therapy. Substrate reduction therapy (SRT) could be of potential benefit. The imino sugars N-butyldeoxynojirimycin (NB DNJ, miglustat, Zavesca) and N-butyldeoxygalactonojirimycin (NB-DGJ) used for SRT inhibit glucosylceramide synthase (GlcCerS) that catalyses the first committed step in glycosphingolipid biosynthesis. We have compared the efficacy and tolerability of NB-DNJ and NB-DGJ in the beta-galactosidase knockout mouse. NB DGJ was better tolerated than NB-DNJ, due to intrinsic gastrointestinal tract dysfunction that was exacerbated by NB-DNJ. However, functional improvement was greatest with NB-DNJ treatment which may potentially be caused by novel anti inflammatory properties of NB-DNJ. PMID- 18387329 TI - Osteoclastic-like giant cell tumour in a cat. AB - An 11-year-old neutered male domestic shorthair cat was presented with an epulis. A hemispherical mass, 8mm in maximum diameter, without a peduncle and bright reddish in colour, was observed on the gingiva of the left mandible. Radiography failed to show any infiltrating osteolysis. The epulis was surgically removed via gingival incision around the margin to the depth of connective tissue layer. Histopathological examination indicated that the epulis contained a large number of multinucleated giant cells (MGCs) intermixed with mononuclear mesenchymal cells in a loose fibrovascular stroma. Mitotic cells were found, mainly in the centre of the mass. MGCs were stained positive by the tartrase resistant acid phosphatase (TRAP) staining, indicating osteoclasts activity. Immunohistochemical staining for proliferating-cell nuclear antigen (PCNA) was observed within the majority of mononucleated cells, whereas multinucleated cells did not stain. An osteoclast-like giant cell tumour was concluded in this case. The origin of epulis is likely from the periosteal tissue. The cat recovered uneventfully and no recurrence has been noted for 3 years thereafter. PMID- 18387330 TI - Cerebellar hemorrhage as an atypical complication of meningococcal meningitis. PMID- 18387331 TI - Ultrasound-assisted synthesis of pyrroles catalyzed by zirconium chloride under solvent-free conditions. AB - A straightforward synthesis of substituted pyrroles using zirconium chloride catalyzed modified Paal-Knorr method has been accomplished under ultrasound irradiation. Compared to known methods, this new method provides as easy access to various substituted pyrroles in good to excellent yields with short reaction times. PMID- 18387332 TI - Perceptions and use of passive intervertebral motion assessment of the spine: a survey among physiotherapists specializing in manual therapy. AB - Manual therapists commonly use passive intervertebral motion (PIVM) assessment within physical examination. Data describing the use and interpretation of this manual diagnostic procedure, as well as therapists' perception of related importance and confidence, are lacking. A survey was conducted among Dutch physiotherapists specializing in manual therapy (MT) using a 13-item, self administered, structured questionnaire. Three hundred and sixty-seven questionnaires were analysed. Response rate from the postal part of the survey was 56%. Dutch manual therapists most frequently apply passive segmental motion assessment to the cervical region and they prefer three-dimensionally coupled motions. They consider end-feel or, to a lesser extent, provocation of patient's pain as decisive for diagnostic conclusions. Respondents believe that these spinal motion tests are important for treatment decisions and are confident in their conclusions drawn from it. These perceptions were largely stable across subgroups of therapists with different gender, age, experience, and educational background. Weekly amount of work related to spinal disorders was positively associated with perceived importance and confidence. Reported use and interpretation of PIVM assessment and related perceptions could only partly be substantiated by evidence. Results from this survey will help researchers design studies better reflecting daily practice in MT. PMID- 18387333 TI - ADAM15 suppresses cell motility by driving integrin alpha5beta1 cell surface expression via Erk inactivation. AB - Human ADAM15 is unique among the A disintegrin and metalloprotease domain (ADAM) family because of the integrin binding motif Arg-Gly-Asp (RGD) within its disintegrin domain. Integrin alpha5beta1 has been reported to bind to ADAM15 in an RGD-dependent manner, but the biological significance of the interaction between ADAM15 and alpha5beta1 is unknown. To characterize the effects of ADAM15 on alpha5beta1-mediated cell adhesion and migration and elucidate the potential mechanism, CHO cells which express endogenous integrin alpha5beta1 were transfected with human ADAM15 cDNA. ADAM15 overexpression led to enhanced cell adhesion and decreased migration on fibronectin, which were suppressed by down regulation of integrin alpha5. Overexpression of ADAM15 not only increased the cell surface expression of integrin alpha5 but also resulted in a more clustered staining of alpha5 on cell surface, while the beta1 subunit remained unchanged. Unexpectedly, results from immunoprecipitation and immunofluorescence indicated that ADAM15 and alpha5beta1 integrin did not interact directly in CHO cells. We found that ADAM15 expression decreased the phosphorylation of Erk1/2. Consistently, down-regulation of Erk1/2 phosphorylation by MEK inhibitor PD98059 or siRNA against Erk1/2 enhanced the expression of alpha5 on cell surface. By using a B16F10 pulmonary metastasis model, we revealed that overexpression of ADAM15 significantly reduced the number of metastatic nodules on the lung. Taken together, this study reveals for the first time that ADAM15 could drive alpha5 integrin expression on cell surface via down-regulation of phosphorylated Erk1/2. This presents a novel mechanism by which ADAM15 regulates cell-matrix adhesion and migration. PMID- 18387334 TI - Induction of a cell-survival adaptive response in MRC-5 cells by hydroquinone. AB - Although it is known that some cell types exhibit an adaptive response to low levels of cytotoxic agents, its molecular mechanism is still unclear and it has yet to be established whether this is a universal phenomenon that occurs in all cell types in response to exposure to every chemical. Hydroquinone is a synthetically produced as well as naturally occurring chemical. Human exposure to hydroquinone is predominantly through diet, cigarette smoke and occupational contact. Here, we asked whether exposure of human lung embryonic MRC-5 fibroblasts to low doses of hydroquinone leads to a cell-survival adaptive response. We further examined the possible mechanisms of an adaptive response using proteomics. We found that exposure of MRC-5 cells to low levels of hydroquinone resulted in adaptation to further exposure to lethal doses of hydroquinone at the cell-survival level, measured using the alamarBlue assay, lactate dehydrogenase leakage assay and Annexin V-FITC/PI staining. To determine the polypeptide products involved in the adaptive response, two-dimensional electrophoresis combined with mass spectrometry was performed. Twenty-three protein spots were significantly changed during the adaptive response. Among them, 21 protein spots were identified by peptide mass fingerprinting and/or peptide sequence analysis by MALDI-TOF-TOF. The identified proteins included proteins involved in energy metabolism, protein folding, redox regulation, cell structure and cell signaling. Our data suggest that the hydroquinone-induced adaptive response is a complex process involving in a modulation of diverse cellular functions, and that the redox regulation might be a common mechanism during the adaptive response. PMID- 18387335 TI - From square to cube: ultrasound as a natural complement of neurophysiology. PMID- 18387336 TI - The ultrasonographic wrist-to-forearm median nerve area ratio in carpal tunnel syndrome. AB - OBJECTIVE: Peripheral nerve ultrasound is an emerging tool in the diagnosis of carpal tunnel syndrome (CTS). Although numerous publications have cited an increased median nerve area at the wrist to be the diagnostic of CTS, there has been considerable variability in the published normal values for this measurement. Our objective is to collect data on the wrist-to-forearm ratio (WFR) of median nerve area in patients with CTS and healthy controls. METHODS: Patients with electrodiagnostically proven CTS underwent ultrasonography of the median nerve at the wrist and forearm. The median nerve area was measured at these points and compared to values from asymptomatic volunteers. RESULTS: The WFR of median nerve area in asymptomatic volunteers was 1.0+/-0.1. The WFR in patients presenting with CTS was 2.1+/-0.5. CONCLUSIONS: The WFR in patients with CTS is elevated as compared to asymptomatic controls. A WFR of 1.4 gave 100% sensitivity for detecting patients with CTS while using only median nerve area at the wrist resulted in a sensitivity of 45-93%, depending on the cut-off value used. SIGNIFICANCE: The WFR of median nerve area promises to be a valid means of diagnosing CTS, and may be superior to measuring median nerve area at the wrist alone. PMID- 18387337 TI - Neuropathic symptoms and findings in women with Fabry disease. AB - OBJECTIVE: To examine the neurologic and neurophysiologic findings and neurologic symptoms in 12 women with Fabry disease and to study the relationship between the subjective symptoms and the findings on the various tests done. METHODS: Neurography, vibratory and thermal quantitative sensory testing (QST), skin biopsy for measuring intraepidermal nerve fiber density (IENFD). Heart rate variability (HRV) and sympathetic skin response (SSR) tests for detecting autonomic dysfunction, pain-, depression- and somatic symptom questionnaires and clinical examination. RESULTS: Only two women had no persistent symptoms or signs of polyneuropathy, 10 had symptoms of small fiber neuropathy. Neurological examination was normal in most patients. Five patients had decreased IENFD or thermal hypoesthesia in QST. In QST, Adelta-fiber function for innocuous cold was more often impaired than C-fiber function. Conventional nerve conduction studies were mostly normal. Carpal tunnel syndrome (CTS) incidence was increased, 25% had symptomatic CTS. CONCLUSIONS: Heterozygous women carrying the gene for Fabry disease have symptoms and findings of small-fiber polyneuropathy more often than has previously been considered. The prevalence of CTS is also increased. SIGNIFICANCE: While the clinical diagnosis of small-fiber neuropathy is difficult, the diagnostic yield can be increased using a combination of thermal QST and IENFD measurements. PMID- 18387338 TI - Difference in intracortical inhibition of the motor cortex between cortical myoclonus and focal hand dystonia. AB - OBJECTIVE: The short interval intracortical inhibition (SICI) of the motor cortex (M1) is reduced in both cortical myoclonus and focal hand dystonia. This reduction has been attributed to the dysfunction of GABAergic system within the motor cortex. However, the precise mechanisms underlying the reduction may not be entirely identical in these two disorders, being due to primary pathological involvement in M1 or secondary to functional changes outside M1. The aim of this study was to elucidate possible differences in intracortical inhibition between these two disorders. METHODS: Subjects were 11 patients with benign myoclonus epilepsy, 7 with focal hand dystonia, and 11 normal volunteers. We studied SICI using anterior-posterior (AP) directed and posterior-anterior (PA) directed induced currents in the brain. RESULTS: In both disorders, SICI with PA-directed currents was reduced as reported previously. In contrast, SICI studied with AP currents was normal in patients with focal hand dystonia, but reduced in patients with cortical myoclonus. CONCLUSIONS: The difference between the two disorders might reflect the underlying pathological difference. In cortical myoclonus, the inhibitory interneurons of the motor cortex are affected, whereas the same interneurons are intact in dystonia. The difference in SICI induced by AP and PA directed currents in dystonia may be explained by the following possibilities: the difference in composition of I-waves contributing to EMG generation and the difference in modulation of the interneuronal activity by voluntary contraction. These changes may be secondary to dysregulation of the motor cortex by the basal ganglia or related cortices in dystonia. SIGNIFICANCE: The SICI using AP directed currents together with the conventional SICI using PA directed currents was able to demonstrate some difference in the intrinsic circuits of M1 between myoclonus and focal hand dystonia. SICI using AP directed currents can provide additional information about the motor cortical excitability changes over those obtained by the previously reported methods. PMID- 18387339 TI - Muscle fiber conduction abnormalities in early diabetic polyneuropathy. AB - OBJECTIVE: Diabetic polyneuropathy (PNP) has been proposed to be a primary disorder of sensory nerves. At an early stage motor nerve conduction velocity (MNCV) and muscle strength remain preserved due to compensatory mechanisms (axonal sprouting, reinnervation). We evaluated the use of invasive muscle fiber conduction velocity (MFCV) measurements as a method to detect muscle fiber denervation atrophy, as an early sign of motor axonal loss in diabetes mellitus (DM). METHODS: Twelve selected male patients (8 type 1, 4 type 2; mean age 35.8 years, SD 10.6), without any sign of micro- or macroangiopathy, were studied by systematic clinical and neurophysiological testing including MFCV estimation. RESULTS: Hand-held dynamometry was normal in all subjects. There were no signs of recent denervation by concentric needle EMG in any of the patients. Sensory nerve conduction velocity (SNCV) was abnormal in 6 subjects, MFCV in 6 subjects (5 had also low SNCV). The ratio of fastest/slowest muscle fibers in MFCV was correlated to SNCV of sural nerve (-.59, p < .05), but not to MNCV. CONCLUSIONS: Half of the clinically asymptomatic DM subjects showed sensory involvement together with MFCV abnormalities, despite normal needle EMG and force. SIGNIFICANCE: MFCV estimation offers a sensitive method in detecting early signs of motor axonal dysfunction in DM. PMID- 18387340 TI - Skull thickness and magnitude of EEG alpha activity. AB - OBJECTIVE: Electroencephalographic (EEG) research on the physiological basis of individual differences in personality or intelligence commonly presumes that between-subjects differences of EEG alpha activity reflect individual differences in brain functions. However, non-functional sources of variance such as individual differences in skull thickness may significantly contribute to individual differences in the magnitude of EEG amplitudes. Aim of the present study was to assess the association between skull thickness and the magnitude of EEG alpha activity. METHODS: A 58-channel EEG was recorded from 49 subjects in resting states at three occasions of measurement each 5 weeks apart. Skull thickness was assessed with proton-weighted images of the head that were acquired with a 1.5 T magnetic resonance imaging (MRI) scanner. RESULTS: There was only a mediocre association between EEG alpha power at frontal, temporal, and parietal sites and the thickness of the underlying skull, with correlations ranging between r = -.36 and r = .10. CONCLUSIONS: This finding suggests that intracranial sources contribute much more variance to the surface EEG than do variations in skull thickness. SIGNIFICANCE: Skull thickness may be neglected as a potent source of error when individual differences in brain activity are indexed by the magnitude of EEG alpha activity. PMID- 18387341 TI - Prevalence and risk factors for obstructive sleep apnea in a multiethnic population of patients presenting for bariatric surgery in Singapore. AB - INTRODUCTION: Obesity is becoming more prevalent world wide. Bariatric surgery is one treatment option for patients with severe or morbid obesity. There have been few comprehensive studies examining prevalence and risk factors for obstructive sleep apnea (OSA) in the multiracial Singaporean bariatric surgery population. METHODS: We performed full polysomnography on 176 consecutive patients undergoing assessment for bariatric surgery. Questionnaires regarding snoring, the presence of witnessed apneas and the Epworth Sleepiness Scale (ESS) were administered. Anthropometric and demographic measurements include age, sex, race, body mass index (BMI) and neck circumference. RESULTS: The prevalence of OSA was 72%, and 49% of the 176 patients had an AHI >= 15. There was a male predominance of OSA (X(2) = 29.7; p<0.001). OSA subjects had larger neck circumference (43.9 +/- 4.5 vs. 39.4 cm +/- 3.3; p<0.001) and higher BMI (43.1 +/- 7.6 vs. 39.1 +/- 5.4 kg/m(2); p<0.001). The neck circumference (OR = 1.37; p<0.001), presence of snoring (OR = 8.25; p<0.001) and an ESS >10 (OR = 3.24; p = 0.03) were significant independent predictors of an AHI >= 15. A neck circumference of 43 cm had an 80% sensitivity and 83% specificity for predicting an AHI >= 15. CONCLUSIONS: OSA is common amongst Singaporeans undergoing evaluation for bariatric surgery, with a high prevalence of moderate and severe disease. An increased neck circumference is a strong independent predictor for an AHI >= 15, with a neck circumference of greater than 43 cm being a sensitive and specific predictor. Race was not found to be a risk factor. PMID- 18387342 TI - Comparing three morningness scales: age and gender effects, structure and cut-off criteria. AB - OBJECTIVE: To add to the validity of the French version of the Composite Scale of Morningness (CSM) by comparing its structure with that of the Morningness Eveningness Questionnaire (MEQ), the most widely used scale to measure circadian typology. Second, to compare the cut-off criteria used to transform the continuous scores into categorical chronotypes. Third, to further test the effects of age and gender on morningness scores. The rMEQ, a shortened version of the MEQ, is also considered. METHODS: Four hundred and fifty-six students served as volunteer subjects and filled the CSM and the MEQ. RESULTS: There was no effect of gender, and the CSM and MEQ scores correlated above (+) 0.90 in both genders. Regarding age, morningness was stable before age 35 and increased afterwards. We replicated the three-factor structure of the CSM previously reported in five different cultures. The MEQ is longer and counted a fourth factor while the first three factors were quite identical to those extracted from the CSM. This comparative study emphasizes the recurrent problem of cut-off scores: the available values for both instruments result in a very poor concordance of chronotypes. CONCLUSIONS: Belonging to the evening-type is regarded as a risk factor for sleep disorders and its positive predictive value should be correctly assessed. Hence, normative scores standardized in such a way they reflect the effects of age, gender and culture are needed for the total score and the factor scores. To this extent, T-scores (with a mean of 50 and standard deviation of 10) would be suitable, and normative tables for French subjects (N = 1598) are given as supplemental data. PMID- 18387343 TI - The diagnosis and management of chronic rheumatic heart disease--an Australian guideline. PMID- 18387345 TI - The NER protein Rad33 shows functional homology to human Centrin2 and is involved in modification of Rad4. AB - In the yeast Saccharomyces cerevisiae the Rad4-Rad23 complex is implicated in the initial damage recognition of the Nucleotide Excision Repair (NER) pathway. NER removes a variety of lesions via two subpathways: Transcription Coupled Repair (TCR) and Global Genome Repair (GGR). We previously showed that the new NER protein Rad33 is involved in both NER subpathways TCR and GGR. In the present study we show UV induced modification of Rad4 that is strongly increased in cells deleted for RAD33. Modification of Rad4 in rad33 cells does not require the incision reaction but is dependent on the TCR factor Rad26. The predicted structure of Rad33 shows resemblance to the Centrin homologue Cdc31. In human cells, Centrin2 binds to XPC and is involved in NER. We demonstrate that Rad4 binds Rad33 directly and via the same conserved amino acids required for the interaction of XPC with Centrin2. Disruption of the Rad4-Rad33 interaction is sufficient to enhance the modification of Rad4 and results in a repair defect similar to that of a rad33 mutant. The current study suggests that the role of Rad33 in the Rad4-Rad23 complex might have parallels with the role of Centrin2 in the XPC-HHR23B complex. PMID- 18387344 TI - Mutations to Ku reveal differences in human somatic cell lines. AB - NHEJ (non-homologous end joining) is the predominant mechanism for repairing DNA double-stranded breaks in human cells. One essential NHEJ factor is the Ku heterodimer, which is composed of Ku70 and Ku86. Here we have generated heterozygous loss-of-function mutations for each of these genes in two different human somatic cell lines, HCT116 and NALM-6, using gene targeting. Previous work had suggested that phenotypic differences might exist between the genes and/or between the cell lines. By providing a side-by-each comparison of the four cell lines, we demonstrate that there are indeed subtle differences between loss-of function mutations for Ku70 versus Ku86, which is accentuated by whether the mutations were derived in the HCT116 or NALM-6 genetic background. Overall, however, the phenotypes of the four lines are quite similar and they provide a compelling argument for the hypothesis that Ku loss-of-function mutations in human somatic cells result in demonstrable haploinsufficiencies. Collectively, these studies demonstrate the importance of proper biallelic expression of these genes for NHEJ and telomere maintenance and they provide insights into why these genes are uniquely essential for primates. PMID- 18387346 TI - Contraceptive practices in women with cystic fibrosis. AB - There is no literature regarding the contraception practices in female CF patients. A chart review identified 69 women with CF of whom two thirds were using contraception. Eleven different forms of contraception were being used with the oral contraceptive pill the preferred method. Despite theoretical concerns regarding efficacy and toxicity, the choices of contraception are similar to those of the general U.S. population. PMID- 18387347 TI - Source of human ventilatory chaos: lessons from switching controlled mechanical ventilation to inspiratory pressure support in critically ill patients. AB - Ventilatory flow measured at the airway opening in humans exhibits a complex dynamics that has the features of chaos. Currently available data point to a neural origin of this feature, but the role of respiratory mechanics has not been specifically assessed. In this aim, we studied 17 critically ill mechanically ventilated patients during a switch form an entirely machine-controlled assistance mode (assist-controlled ventilation ACV) to a patient-driven mode (inspiratory pressure support IPS). Breath-by-breath respiratory variability was assessed with the coefficient of variation of tidal volume, total cycle time, inspiratory time, expiratory time, mean inspiratory flow, duty cycle. The detection of chaos was performed with the noise titration technique. When present, chaos was characterized with numerical indexes (correlation dimension, irregularity; largest Lyapunov exponent, sensitivity to initial conditions). Expectedly, the coefficients of variations of the respiratory variables were higher during IPS than during ACV. During ACV, noise titration failed to detect nonlinearities in 12 patients who did not exhibit signs of spontaneous respiratory activity. This indicates that the mechanical properties of the respiratory system were not sufficient to produce ventilatory chaos in the presence of a nonlinear command (ventilator clock). A positive noise limit was found in the remaining 5 cases, but these patients exhibited signs of active expiratory control (highly variable expiratory time, respiratory frequency higher than the set frequency). A positive noise limit was also observed in 16/17 patients during IPS (p<0.001). These observations suggest that ventilatory chaos predominantly has a neural origin (intrinsic to the respiratory central pattern generators, resulting from their perturbation by respiratory afferents, or both), with little contribution of respiratory mechanics, if any. PMID- 18387348 TI - Layer-by-layer assembly of viral capsid for cell adhesion. AB - Cowpea mosaic virus (CPMV)-based thin films are biologically active for cell culture. Using layer-by-layer assembly of CPMV and poly(diallyldimethylammonium chloride), quantitatively scalable biomolecular surfaces were constructed, which were well characterized using quartz crystal microbalance, UV-vis and atomic force microscopy. The surface coverage of CPMV nanoparticles depended on the adsorption time and pH of the virus solution, with a greater amount of CPMV adsorption occurring near its isoelectric point. It was found that the adhesion and proliferation of NIH-3T3 fibroblasts can be controlled by the coverage of viral particles using this multilayer technique. PMID- 18387349 TI - Bone tissue engineering with premineralized silk scaffolds. AB - Silk fibroin biomaterials are being explored as novel protein-based systems for cell and tissue culture. In the present study, biomimetic growth of calcium phosphate on porous silk fibroin polymeric scaffolds was explored to generate organic/inorganic composites as scaffolds for bone tissue engineering. Aqueous derived silk fibroin scaffolds were prepared with the addition of polyaspartic acid during processing, followed by the controlled deposition of calcium phosphate by exposure to CaCl(2) and Na(2)HPO(4). These mineralized protein composite scaffolds were subsequently seeded with human bone marrow stem cells (hMSC) and cultured in vitro for 6 weeks under osteogenic conditions with or without BMP-2. The extent of osteoconductivity was assessed by cell numbers, alkaline phosphatase and calcium deposition, along with immunohistochemistry for bone-related outcomes. The results suggest increased osteoconductive outcomes with an increase in initial content of apatite and BMP-2 in the silk fibroin porous scaffolds. The premineralization of these highly porous silk fibroin protein scaffolds provided enhanced outcomes for the bone tissue engineering. PMID- 18387351 TI - Intermittent Fugu parathyroid hormone 1 (1-34) is an anabolic bone agent in young male rats and osteopenic ovariectomized rats. AB - Human parathyroid hormone (hPTH) is currently the only treatment for osteoporosis that forms new bone. Previously we described a fish equivalent, Fugu parathyroid hormone 1 (fPth1) which has hPTH-like biological activity in vitro despite fPth1(1-34) sharing only 53% identity with hPTH(1-34). Here we demonstrate the in vivo actions of fPth1(1-34) on bone. In study 1, young male rats were injected intermittently for 30 days with fPth1 [30 microg-1,000 microg/kg body weight (b.w.), (30fPth1-1,000fPth1)] or hPTH [30 microg-100 microg/kg b.w. (30hPTH 100hPTH)]. In proximal tibiae at low doses, the fPth1 was positively correlated with trabecular bone volume/total volume (TbBV/TV) while hPTH increased TbBV/TV, trabecular thickness (TbTh) and trabecular number (TbN). 500fPth1 and 1000fPth1 increased TbBV/TV, TbTh, TbN, mineral apposition rate (MAR) and bone formation rate/bone surface (BFR/BS) with a concomitant decrease in osteoclast surface and number. In study 2 ovariectomized (OVX), osteopenic rats and sham operated (SHAM) rats were injected intermittently with 500 microg/kg b.w. of fPth1 (500fPth1) for 11 weeks. 500fPth1 treatment resulted in increased TbBV/TV (151%) and TbTh (96%) in the proximal tibiae due to increased bone formation as assessed by BFR/BS (490%) and MAR (131%). The effect was restoration of TbBV/TV to SHAM levels without any effect on bone resorption. 500fPth1 also increased TbBV/TV and TbTh in the vertebrae (L6) and cortical thickness in the mid-femora increasing bone strength at these sites. fPth1 was similarly effective in SHAM rats. Notwithstanding the low amino acid sequence homology with hPTH (1-34), we have clearly established the efficacy of fPth1 (1-34) as an anabolic bone agent. PMID- 18387350 TI - Interaction between human umbilical vein endothelial cells and human osteoprogenitors triggers pleiotropic effect that may support osteoblastic function. AB - Osteogenesis occurs in striking interaction with angiogenesis. There is growing evidence that endothelial cells are involved in the modulation of osteoblast differentiation. We hypothesized that primary human umbilical vein endothelial cells (HUVEC) should be able to modulate primary human osteoprogenitors (HOP) function in an in vitro co-culture model. In a previous study we demonstrated that a 3 day to 3 week co-culture stimulates HOP differentiation markers such as Alkaline Phosphatase (ALP) activity and mineralization. In the present study we addressed the effects induced by the co-culture on HOP within the first 48 hours. As a prerequisite, we validated a method based on immuno-magnetic beads to separate HOP from HUVEC after co-culture. Reverse transcription-real time quantitative PCR studies demonstrated up-regulation of the ALP expression in the co-cultured HOP, confirming previous results. Surprisingly, down-regulation of runx2 and osteocalcin was also shown. Western blot analysis revealed co-culture induced down-regulation of Connexin43 expression in both cell types. Connexin43 function may be altered in co-cultured HOP as well. Stimulation of the cAMP pathway was able to counterbalance the effect of the co-culture on the ALP activity, but was not able to rescue runx2 mRNA level. Co-culture effect on HOP transcriptome was analyzed with GEArray cDNA microarray showing endothelial cells may also modulate HOP extracellular matrix production. In accordance with previous work, we propose endothelial cells may support initial osteoblastic proliferation but do not alter the ability of the osteoblasts to produce extracellular mineralizing matrix. PMID- 18387353 TI - Development of a one-step strip for the detection of triazophos residues in environmental samples. AB - Environmental and food safety issues now are recognized internationally, and pesticide residues play key roles as environment and food pollutants. It is crucial to develop methods for rapid determination of pesticide residues in environments and foods. A one-step strip based on nanocolloidal-gold-labeled monoclonal antibodies for detection of triazophos residue was developed. The nanocolloidal gold, with an average particle diameter of 25 nm (G25), was labeled to an antitriazophos monoclonal antibody. This conjugate was dispensed on the conjugate pad of a porous glass fiber. Ovalbumin hapten and goat anti-mouse IgG were dispensed on the nitrocellulose membrane and served as the test line (T line) and control line (C-line), respectively. After conditions optimization, the one-step strip was finally developed for the residue determination of triazophos. The limit of detection (LOD) of the strip was 4 ng/mL for standard. The detection was not affected by the pH of the liquid sample but low total ion concentration will induce illegible C-line and T-line. The LOD for spiked samples of soil and water was 5ng/mL, with run time of no more than 10min. PMID- 18387352 TI - A comparison of sugar indicators enables a universal high-throughput sugar-1 phosphate nucleotidyltransferase assay. AB - A systematic comparison of six sugar indicators for their sensitivity, specificity, cross-reactivity, and suitability in the context of crude lysates revealed para-hydroxybenzoic acid hydrazide (pHBH) to be best suited for application in a plate-based phosphatase-assisted universal sugar-1-phosphate nucleotidyltransferase assay. The addition of a general phosphatase to nucleotidyltransferase reaction aliquots enabled the conversion of remaining sugar-1-phosphate to free sugar, the concentration of which could be rapidly assessed via the pHBH assay. The assay was validated using the model glucose-1 phosphate thymidylyltransferase from Salmonella enterica (RmlA) and compared favorably with a previously reported HPLC assay. This coupled discontinuous assay is quantitative, high throughput, and robust; relies only on commercially available enzymes and reagents; does not require chromatography, specialized detectors (e.g., mass or evaporative light scattering detectors), or radioisotopes; and is capable of detecting less than 5 nmol of sugar-1-phosphate. It is anticipated that this high-throughput assay system will greatly facilitate nucleotidyltransferase mechanistic and directed evolution/engineering studies. PMID- 18387354 TI - Self-structure induction in single stranded poly(A) by small molecules: Studies on DNA intercalators, partial intercalators and groove binding molecules. AB - Self-structure induction in single stranded poly(A) has been one typical example of the various ways that could be used to modulate nucleic acid structural aspects through binding of small molecules. For the first time, the interaction between a series of small molecules and poly(A) has been investigated to understand the nature of the structural features in DNA binding small molecules that could be responsible for the formation of self-structure in single stranded poly(A) molecules. Classical intercalators like ethidium, coralyne, quinacrine and proflavine, partial intercalators like berberine and palmatine and classical minor groove binders like hoechst 33258 and DAPI have been chosen for this study. The binding of each of these molecules to poly(A) has been characterized by absorption spectral titration, job plot and isothermal titration calorimetry. Self-structure formation was monitored from circular dichroic melting, optical melting and differential scanning calorimetry. The results revealed that while all the intercalators studied induced self-structure formation, partial intercalators did not induce the same in poly(A). Of the two classical DNA minor groove binding molecules investigated, hoechst was effective in inducing self structure while DAPI was ineffective. Self-structure induction in poly(A) was observed to be directly linked to the cooperative binding of the molecules to poly(A) in that all the molecules that bound cooperatively induced self-structure in poly(A). Structural and thermodynamic aspects of the interaction leading to self-structure formation are described. PMID- 18387355 TI - Murine platelets are not regulated by O-linked beta-N-acetylglucosamine. AB - It is generally appreciated that platelets derived from diabetic patients display increased responsiveness to low levels of agonists. O-GlcNAcylation has been linked to hyperglycemia-related effects in other tissues; therefore we examined this modification in platelets to determine if O-GlcNAcylation affects platelet function. This post-translational modification consists of an N-acetylglucosamine attached to serine and/or threonine residues. We examined O-GlcNAc levels in platelets from a hyperglycemic murine model of Type I diabetes with known hypersensitivity to agonists and a Type II diabetes model (ob/ob) lacking detectable alterations in the aggregation profile. Neither model showed marked increases in protein O-GlcNAcylation. Treatment of platelets with multiple O GlcNAcase inhibitors led to O-GlcNAc accumulation on multiple platelet proteins. However, the inhibitor-induced accumulation of this modification does not correlate with any gross alterations in platelet aggregation. These data suggest that while the modification occurs in platelets, their activity is not globally sensitive to O-GlcNAc levels. PMID- 18387356 TI - The energetics of the primary proton transfer in bacteriorhodopsin revisited: it is a sequential light-induced charge separation after all. AB - The light-induced proton transport in bacteriorhodopsin has been considered as a model for other light-induced proton pumps. However, the exact nature of this process is still unclear. For example, it is not entirely clear what the driving force of the initial proton transfer is and, in particular, whether it reflects electrostatic forces or other effects. The present work simulates the primary proton transfer (PT) by a specialized combination of the EVB and the QCFF/PI methods. This combination allows us to obtain sufficient sampling and a quantitative free energy profile for the PT at different protein configurations. The calculated profiles provide new insight about energetics of the primary PT and its coupling to the protein conformational changes. Our finding confirms the tentative analysis of an earlier work (A. Warshel, Conversion of light energy to electrostatic energy in the proton pump of Halobacterium halobium, Photochem. Photobiol. 30 (1979) 285-290) and determines that the overall PT process is driven by the energetics of the charge separation between the Schiff base and its counterion Asp85. Apparently, the light-induced relaxation of the steric energy of the chromophore leads to an increase in the ion-pair distance, and this drives the PT process. Our use of the linear response approximation allows us to estimate the change in the protein conformational energy and provides the first computational description of the coupling between the protein structural changes and the PT process. It is also found that the PT is not driven by twist-modulated changes of the Schiff base's pKa, changes in the hydrogen bond directionality, or other non-electrostatic effects. Overall, based on a consistent use of structural information as the starting point for converging free energy calculations, we conclude that the primary event should be described as a light-induced formation of an unstable ground state, whose relaxation leads to charge separation and to the destabilization of the ion-pair state. This provides the driving force for the subsequent PT steps. PMID- 18387357 TI - GATA-4 modulates C/EBP-dependent transcriptional activation of acute phase protein genes. AB - C/EBP transcription factors are involved in the regulation of the intestinal epithelial cell response to inflammatory stimuli. GATA transcription factors modulate C/EBP-dependent transcriptional activation in various cell types. We thus determined whether GATA-4 whose expression is restricted to epithelial cells, modulate C/EBP transcriptional activity and C/EBP-dependent acute phase protein expression in intestinal epithelial cells. Interaction between C/EBPdelta and GATA-4 required both C/EBPdelta leucine zipper and basic DNA-binding domains, and the GATA-4 C-terminal region. C/EBP isoforms and GATA-4 synergistically activated the alpha-acid glycoprotein gene while GATA-4 repressed thiostatin and haptoglobin C/EBP-dependent transactivation. In GATA-4 expressing intestinal epithelial cells, GATA-4 led to decreased C/EBPbeta and C/EBPdelta protein levels, and decreased thiostatin expression in response to IL-1beta and dexamethasone. This correlated with decreased C/EBPdelta recruitment to the thiostatin promoter, as assessed by chromatin immunoprecipitation assays. In contrast, increased AGP expression in response to dexamethasone correlated with increased basal histone H4 acetylation. Thus, GATA-4 may be involved in the establishment of specific intestinal epithelial cell C/EBP-dependent and C/EBP independent transcriptional responses. PMID- 18387358 TI - Human chromosome 21-derived miRNAs are overexpressed in down syndrome brains and hearts. AB - Down syndrome (DS), or Trisomy 21, is the most common genetic cause of cognitive impairment and congenital heart defects in the human population. To date, the contribution of microRNAs (miRNAs) in DS has not been investigated. Bioinformatic analyses demonstrate that human chromosome 21 (Hsa21) harbors five miRNA genes; miR-99a, let-7c, miR-125b-2, miR-155, and miR-802. MiRNA expression profiling, miRNA RT-PCR, and miRNA in situ hybridization experiments demonstrate that these miRNAs are overexpressed in fetal brain and heart specimens from individuals with DS when compared with age- and sex-matched controls. We hypothesize that trisomic 21 gene dosage overexpression of Hsa21-derived miRNAs results in the decreased expression of specific target proteins and contribute, in part, to features of the neuronal and cardiac DS phenotype. Importantly, Hsa21-derived miRNAs may provide novel therapeutic targets in the treatment of individuals with DS. PMID- 18387359 TI - Uridine diphosphate (UDP) stimulates insulin secretion by activation of P2Y6 receptors. AB - We examined the transcriptional expression and functional effects of receptors for the extracellular pyrimidines uridine triphosphate (UTP) and uridine diphosphate (UDP), on insulin and glucagon secretion in isolated mouse pancreatic islets and purified beta-cells. Using real-time PCR, the UDP receptor P2Y(6) was found to be highly expressed in both whole islets and beta-cells purified by repeated counter-flow elutriation, whereas no mRNA expression for UTP receptors P2Y(4) and P2Y(2) could be detected. Functional in vitro experiments revealed that the P2Y(6) agonist UDPbetaS dose-dependently enhanced insulin and glucagon release during short-term incubation (1h), while P2Y(6) activation during a longer period (24h), selectively increased insulin release, especially at high glucose levels. The corresponding EC(50) value for UDPbetaS ranged from 3.2 x 10( 8)M to 1.6 x 10(-8)M for both glucose concentrations. The P2Y(6) antagonist MRS2578 inhibited the effects of UDPbetaS, supporting a P2Y(6) specific effect. In addition to negative RT-PCR results, the lack of response to UTPgammaS a selective P2Y(2/4) agonist further rule out the involvement of P2Y(2/4) receptors in the islet hormone release. Our results suggest a modulatory role for UDP via a functional active P2Y(6) receptor in the regulation of islet hormone release. PMID- 18387360 TI - Independent and incremental prognostic value of multimarker testing in acute dyspnea: results from the ProBNP Investigation of Dyspnea in the Emergency Department (PRIDE) study. AB - BACKGROUND: Acute dyspnea is common in the emergency department (ED) and is associated with mortality. Biomarkers may help stratify risk in this setting. METHODS: Among 577 dyspneic subjects we identified 5 candidate biomarkers with prognostic value: amino terminal B-type natriuretic peptide (NT-proBNP), C reactive protein (CRP), the interleukin family member ST2, hemoglobin and blood urea nitrogen (BUN); these were assessed using both receiver operating characteristic curve and Cox proportional hazards analyses. Results were validated in a population of dyspneic patients from a distinct cohort. RESULTS: At 1 y follow up, 93 (16.1%) patients had died. Independent predictive ability was established in an age-adjusted Cox model containing all markers: NT-proBNP (HR=1.89); CRP (HR=1.95); ST2 (HR=7.17); hemoglobin (HR=1.68); BUN (HR=2.06) (all P<.05). Following categorical assessment based on number of abnormal markers, the 1-y risk of death increased in a monotonic fashion with mortality rates of 0%, 2.0%, 7.8%, 22.3%, 29.3%, and 57.6% respectively; similar results were seen in the validation set. CONCLUSION: Simultaneous assessment of pathophysiologically diverse markers in acute dyspnea provides powerful, independent and incremental prognostic information. PMID- 18387361 TI - Prolidase activity in serum and pleural fluids in patients with tuberculous pleural effusion [correction of effussion]. AB - OBJECTIVES: Pleural tuberculosis, which is present in around 4% of all tuberculosis cases may resolve spontaneously or associated with progressive disease and a high recurrence rate. Recently upon exposed to cytokines and bacterial products, mesothelium has been shown to produce collagen that may be involved in pleural inflammatory responses. Prolidase is involved in the final stage of degradation in collagen catabolism. In this study we aimed to evaluate pleural fluid and serum prolidase activities in patients with tuberculous (TB) pleurisy and compared with those in non-tuberculous (non-TB) pleural effusions. DESIGN AND METHODS: 21 patients with tuberculous (TB) pleurisy (11 F/10 M), ages 35-52 (median 44) and 22 patients (10 F/12 M), ages 41-63 (median 52) with non tuberculous pleurisy included as non-tuberculous (non-TB) pleurisy group consecutively referred to our pulmonary clinic for evaluation. Serum and pleural prolidase activities in 21 TB and 22 non-TB pleurisy patients were analyzed by photometric method. RESULTS: Prolidase enzyme activities in serum and pleural fluids of TB group (1072+/-171 and 1392+/-215 U/L, respectively) were significantly higher than those values in non-TB group (787+/-144 and 943+/-174 U/L, respectively). Prolidase activities in pleural fluid were significantly higher than those in serum in both groups. There was a significant positive correlation between pleural and serum prolidase activities in TB group (r=0.579 and p=0.006) and in non-TB group (r=0.858 and p<0.001). In Receiver Operating Characteristic (ROC) analysis, sensitivity and specificity values were 86% and 82% for a cut-off value of 1130 U/L for pleural prolidase activity and were 81% and 82% for a cut-off value of 952 U/L for serum prolidase activity, respectively. CONCLUSION: In conclusion, there is an elevated pleural fluid and serum prolidase enzyme activity in patients with TB pleurisy compared with non-TB pleurisy group. The higher enzyme activities in TB group might reflect increased collagen turnover in those patients. PMID- 18387362 TI - Low diurnal variability of apolipoprotein A1, apolipoprotein B and apolipoprotein B/apolipoprotein A1 ratio during normal sleep and after an acute shift of sleep. AB - OBJECTIVE: The aim of this study was to study the diurnal variation of the cardiovascular risk markers apolipoprotein A1 and B and apo B/apo A1 ratio. DESIGN AND METHODS: We have studied the diurnal variation of apolipoprotein A1, apolipoprotein B and apo B/apo A1 ratio during night sleep and the day sleep conditions in seven healthy volunteers (age 22-32 yr). Samples were collected every hour to evaluate the effect of different sampling times on the test results. RESULTS: The lowest diurnal coefficient of variation (CV) was observed for the apo B/apo A1 ratio, which usually was below 2% but also apolipoprotein A1, apolipoprotein B showed low CV. There were no significant differences between nightsleep and daysleep for any of the studied markers. CONCLUSION: Even if there was a diurnal variation for these markers, the variation was very low. Thus, sampling does not have to be restricted to certain times of the day. PMID- 18387363 TI - Analysis of coenzyme Q10 in muscle and fibroblasts for the diagnosis of CoQ10 deficiency syndromes. AB - OBJECTIVES: To study CoQ(10) concentrations in muscle and fibroblast from 6 patients with a CoQ(10) deficiency syndrome. DESIGN AND METHODS: CoQ(10) was quantified by HPLC with electrochemical detection. RESULTS: Four out of the 6 cases showed muscle CoQ(10) deficiency plus a reduction of mitochondrial respiratory chain enzyme activities. All cases showed decreased CoQ(10) values in fibroblasts when compared with controls. CONCLUSIONS: Biochemical study of CoQ(10) in both muscle and fibroblasts seems advisable to demonstrate the deficiency in all patients. PMID- 18387364 TI - The mammalian Nek1 kinase is involved in primary cilium formation. AB - Recent studies implicate primary cilium (PC) proteins in the etiologies of various polycystic kidney diseases (PKD). NIMA-related kinases (NRKs) are conserved serine/threonine kinases, which are usually defined as 'mitotic kinases'. Murine mutants for the NRKs, nek1 (kat mice) suffer from PKD, suggesting that it may be involved in cilium control. We demonstrated herein that Nek1 is localized to basal body region and that Nek1 overexpression inhibits ciliogenesis in Madin-Darby canine kidney epithelial cells. The number of primary cilia is dramatically reduced in kat2J mouse embryonic fibroblasts culture. It is thus hypothesized that Nek1 links cell cycle progression and the PC cycle. PMID- 18387365 TI - Detection of cross-links between FtsH, YidC, HflK/C suggests a linked role for these proteins in quality control upon insertion of bacterial inner membrane proteins. AB - Little is known about the quality control of proteins upon integration in the inner membrane of Escherichia coli. Here, we demonstrate that YidC and FtsH are adjacent to a nascent, truncated membrane protein using in vitro photo cross linking. YidC plays a critical but poorly understood role in the biogenesis of E. coli inner membrane proteins (IMPs). FtsH functions as a membrane chaperone and protease. Furthermore, we show that FtsH and its modulator proteins HflK and HflC copurify with tagged YidC and, vice versa, that YidC copurifies with tagged FtsH. These results suggest that FtsH and YidC have a linked role in the quality control of IMPs. PMID- 18387366 TI - Mapping of interaction domains of putative telomere-binding proteins AtTRB1 and AtPOT1b from Arabidopsis thaliana. AB - We previously searched for interactions between plant telomere-binding proteins and found that AtTRB1, from the single-myb-histone (Smh) family, interacts with the Arabidopsis POT1-like-protein, AtPOT1b, involved in telomere capping. Here we identify domains responsible for that interaction. We also map domains in AtTRB1 responsible for interactions with other Smh-family-members. Our results show that the N-terminal OB-fold-domain of AtPOT1b mediates the interaction with AtTRB1. This domain is characteristic for POT1- proteins and is involved with binding the G-rich-strand of telomeric DNA. AtPOT1b also interacts with AtTRB2 and AtTRB3. The central histone-globular-domain of AtTRB1 is involved with binding to AtTRB2 and 3, as well as to AtPOT1b. AtTRB1-heterodimers with other Smh-family-members are more stable than AtTRB1-homodimers. Our results reveal interaction networks of plant telomeres. PMID- 18387367 TI - Parkin-co-regulated gene (PACRG) product interacts with tubulin and microtubules. AB - Parkin-co-regulated gene (PACRG) is a gene that shares a bidirectional promoter with Parkinson's disease-related Parkin/Park2 gene. Recently, the PACRG gene product was implicated in the function of flagella. However, its exact function remains unknown. Here, I assessed the interaction between PACRG and tubulin. Co sedimentation experiments revealed that PACRG directly binds to microtubules and alpha/beta-tubulin heterodimers with high affinity. Microscopic studies showed that PACRG bundles microtubules and forms branched aggregates with unpolymerized tubulin dimers. The amino acid sequence of the microtubule-binding region of PACRG is highly conserved among various organisms, suggesting that tubulin binding is a basic property of PACRG. PMID- 18387368 TI - Small heat shock protein Hsp27 protects myosin S1 from heat-induced aggregation, but not from thermal denaturation and ATPase inactivation. AB - We applied different methods, such as turbidity measurements, dynamic light scattering, differential scanning calorimetry and co-sedimentation assay, to analyze the interaction of small heat shock protein Hsp27 with isolated myosin head (myosin subfragment 1, S1) under heat-stress conditions. Upon heating at 43 degrees C, Hsp27 effectively suppresses S1 aggregation, and this effect is enhanced by mutations mimicking Hsp27 phosphorylation. However, Hsp27 was unable to prevent thermal unfolding of myosin heads and to maintain their ATPase activity under heat-shock conditions. PMID- 18387370 TI - Dolichol-phosphate mannose synthase: structure, function and regulation. AB - Glycosylation is the major modification of proteins, and alters their structures, functions and localizations. Glycosylation of secretory and surface proteins takes place in the endoplasmic reticulum and Golgi apparatus in eukaryotic cells and is classified into four modification pathways, namely N- and O-linked glycosylations, glycosylphosphatidylinositol (GPI)-anchor and C-mannosylation. These modifications are accomplished by sequential addition of single monosaccharides (O-linked glycosylation and C-mannosylation) or en bloc transfer of lipid-linked oligosaccharides (N-linked glycosylation and GPI) onto the proteins. The glycosyltransferases involved in these glycosylations are categorized into two classes based on the type of sugar donor, namely nucleotide sugars and dolichol-phosphate-sugars, in which the sugar moiety is mannose or glucose. The sugar transfer from dolichol-phosphate-sugars occurs exclusively on the luminal side of the endoplasmic reticulum and is utilized in all four glycosylation pathways. In this review, we focus on the biosynthesis of dolichol phosphate-mannose, and particularly on the mammalian enzyme complex involved in the reaction. PMID- 18387371 TI - Gender and age-dependent differences in the mitochondrial apoptogenic pathway in Alzheimer's disease. AB - Age-related mitochondrial oxidative stress is highly gender dependent. The aim of this study was to determine the role of gender in the mitochondrial contribution to neuronal apoptosis in Alzheimer's disease (AD). We used mitochondria isolated from brains of Wistar rats to study the toxicity of ss-amyloid peptide (Ass), and found that it increases mitochondrial peroxide production, nitration and oxidation of proteins, and release of cytochrome c. The toxic effects occurred in young males and in old females but not in young females, indicating their resistance to Ass. This resistance was abolished with age. These toxic effects of Ass were prevented by heme. Our findings provide a molecular mechanism for the contribution of Abeta to the mitochondrial dysfunction and oxidative stress seen in AD, as well as for the mitochondria-dependent pathway of apoptosis in AD. Gender and age-related differences seen in the development of AD can also be partially explained. PMID- 18387372 TI - Conformational analysis of the broad-spectrum antibacterial peptide, ranatuerin 2CSa: identification of a full length helix-turn-helix motif. AB - Design of clinically valuable antibacterial agents based upon naturally occurring peptides requires the use of spectroscopic methods, particularly NMR, to determine the three-dimensional structure of the native peptide so that analogues with improved therapeutic properties can be made. Ranatuerin-2CSa (GILSSFKGVAKGVAKDLAG KLLETLKCKITGC), first isolated from skin secretions of the Cascades frog, Rana cascadae, represents a promising candidate for drug development. The peptide shows potent growth inhibitory activity against Escherichia coli (MIC=5 microM) and Staphylococcus aureus (MIC=10 microM) but displays haemolytic activity against human erythrocytes (LC(50)=160 microM). The solution structure of ranatuerin-2CSa was investigated by proton NMR spectroscopy and molecular modelling. In aqueous solution, the peptide lacks secondary structure but, in a 2,2,2-trifluoroethanol (TFE-d(3))-H(2)O solvent mixture, the structure is characterised by a full length helix-turn-helix conformation between residues I(2)-L(21), L(22)-L(25) and K(26)-T(30) respectively. This structural information will facilitate the design of novel therapeutic agents based upon the ranatuerin-2CSa structure with improved antimicrobial potencies but decreased cytolytic activities against mammalian cells. PMID- 18387373 TI - Medical clinics of North America. Common gastrointestinal emergencies. Preface. PMID- 18387374 TI - Initial management of acute upper gastrointestinal bleeding: from initial evaluation up to gastrointestinal endoscopy. AB - Acute upper gastrointestinal bleeding is a relatively common, potentially life threatening medical emergency responsible for more than 300,000 hospital admissions and about 30,000 deaths per annum in America. The initial assessment focuses on bleeding activity, bleeding severity, hemodynamic compromise from the bleeding, and differentiating upper from lower gastrointestinal bleeding. The initial supportive therapy includes fluid resuscitation to reverse the hypovolemia, blood transfusions to replete the lost blood, respiratory support as necessary, and proton pump inhibitor therapy to stabilize mucosal blood clots and promote hemostasis. Esophagogastroduodenoscopy is the best test to determine the bleeding site and cause. PMID- 18387375 TI - Acute nonvariceal upper gastrointestinal bleeding: endoscopic diagnosis and therapy. AB - Acute upper gastrointestinal bleeding is a relatively common,potentially life threatening condition that causes more than 300,000 hospital admissions and about 30,000 deaths per annum in America. Esophagogastroduodenoscopy is the procedure of choice for the diagnosis and therapy of upper gastrointestinal bleeding lesions. Endoscopic therapy is indicated for lesions with high risk stigmata of recent hemorrhage, including active bleeding, oozing, a visible vessel, and possibly an adherent clot. Endoscopic therapies include injection therapy, such as epinephrine or sclerosant injection; ablative therapy, such as heater probe or argon plasma coagulation; and mechanical therapy, such as endoclips or endoscopic banding. Endoscopic therapy reduces the risk of rebleeding,the need for blood transfusions, the requirement for surgery, and patient morbidity. PMID- 18387376 TI - Portal hypertension and variceal hemorrhage. AB - Portal hypertension, a major hallmark of cirrhosis, is defined as a portal pressure gradient exceeding 5 mm Hg. In portal hypertension, porto-systemic collaterals decompress the portal circulation and give rise to varices. Successful management of portal hypertension and its complications requires knowledge of the underlying pathophysiology, the pertinent anatomy, and the natural history of the collateral circulation, particularly the gastroesophageal varices. PMID- 18387369 TI - Biosynthesis and function of polyacetylenes and allied natural products. AB - Polyacetylenic natural products are a substantial class of often unstable compounds containing a unique carbon-carbon triple bond functionality, that are intriguing for their wide variety of biochemical and ecological functions, economic potential, and surprising mode of biosynthesis. Isotopic tracer experiments between 1960 and 1990 demonstrated that the majority of these compounds are derived from fatty acid and polyketide precursors. During the past decade, research into the metabolism of polyacetylenes has swiftly advanced, driven by the cloning of the first genes responsible for polyacetylene biosynthesis in plants, moss, fungi, and actinomycetes and the initial characterization of the gene products. The current state of knowledge of the biochemistry and molecular genetics of polyacetylenic secondary metabolic pathways will be presented together with an up-to-date survey of new terrestrial and marine natural products, their known biological activities, and a discussion of their likely metabolic origins. PMID- 18387377 TI - Mechanical obstruction of the small bowel and colon. AB - Mechanical obstruction of the small bowel and colon is moderately common, accounting for several hundred thousand admissions per year in the United States. Patients generally present with abdominal pain, nausea and emesis, abdominal distention, and progressive obstipation. Clinical findings of high fever, localized severe abdominal tenderness, rebound tenderness, severe leukocytosis, or metabolic acidosis suggest possible complications of bowel necrosis, bowel perforation, or generalized peritonitis. Differentiation of total mechanical obstruction from partial mechanical obstruction and pseudo-obstruction is important because total mechanical obstruction is generally treated surgically,whereas the other two entities are usually treated medically. Mechanical obstruction is usually suggested by plain abdominal radiographs, and confirmed by small bowel follow through,abdominal CT, or CT enteroclysis. PMID- 18387378 TI - Gastrointestinal perforation and the acute abdomen. AB - The acute abdomen accounts for up to 40% of all emergency-surgical hospital admissions and is considered in the differential in the more than 7 million visits to the emergency department annually for abdominal pain in the United States. A large percentage of these cases are secondary to perforation or impending gastrointestinal perforation. Gastrointestinal perforation causes considerable mortality and usually requires emergency surgery.Rapid diagnosis and treatment of these conditions is essential to reduce the high morbidity and mortality of late-stage presentation. Successful treatment requires a thorough understanding of the anatomy, microbiology, and pathophysiology of this disease process and in-depth knowledge of the therapy, including resuscitation,antibiotics, source control, and physiologic support. PMID- 18387379 TI - Acute abdominal vascular emergencies. AB - Abdominal vascular emergencies are relatively uncommon, frequently catastrophic, and highly lethal. Despite improved understanding of the pathophysiology and natural history of these disorders, delays in diagnosis and treatment remain the most important factors contributing to the observed high mortality. A high index of clinical suspicion together with a sound understanding of the clinical presentation, natural history, and management of these disorders are critical to improving outcomes. This article focuses on abdominal vascular emergencies presenting with acute visceral ischemia or catastrophic intra-abdominal hemorrhage. PMID- 18387380 TI - Adynamic ileus and acute colonic pseudo-obstruction. AB - Ileus and colonic pseudo-obstruction cause functional obstruction of intestinal transit, without mechanical obstruction, because of uncoordinated or attenuated intestinal muscle contractions. Ileus usually arises from an exaggerated intestinal reaction to abdominal surgery that is often exacerbated by numerous other conditions. Colonic pseudo-obstruction is induced by numerous metabolic disorders, drugs that inhibit intestinal motility, severe illnesses, and extensive surgery. It presents with massive colonic dilatation with variable, moderate small bowel dilatation. Both conditions are initially treated with supportive measures that include intravenous rehydration, correction of electrolyte abnormalities, discontinuation of antikinetic drugs, and treatment of other contributing disorders. Specific therapies for colonic pseudo-obstruction include neostigmine (an anticholinesterase) for pharmacologic colonic decompression and colonoscopic decompression. PMID- 18387381 TI - Severe complications of inflammatory bowel disease. AB - Patients who have inflammatory bowel disease occasionally develop severe complications or emergency situations that require expert and expedited medical care, including toxic colitis, fistulas, abdominal abscesses, malignancy, primary sclerosing cholangitis, and pouchitis. Morbidity and mortality rates of Crohn's disease and ulcerative colitis are increased over the expected rates in the unaffected population. Knowledge of the presenting features, natural history, and treatment of these complications should to lead to early and effective therapy and better outcomes. PMID- 18387382 TI - Endoscopic mucosal resection of colonic lesions: current applications and future prospects. AB - The introduction of submucosal fluid injection has remarkably extended the range of endoscopically resectable polyps. The limiting factor for endoscopic resection is not polyp size, but polyp depth. Endoscopic ultrasound is a useful adjunctive diagnostic tool to assess the depth of invasion. The success of are section ultimately depends on pathologic confirmation of a benign nature of this lesion or of a cancer limited to the mucosa. Selected well-differentiated cancers without lymphovascular invasion of the superficial submucosa can be successfully resected endoscopically. PMID- 18387383 TI - What every graduating medical student should know about urology: the stakeholder viewpoint. AB - OBJECTIVES: What should every medical student know about urology upon graduating from medical school? To answer this question, we conducted a survey of key stakeholders in an effective medical student education in urology. METHODS: Directors of the generalist residencies in the United States (emergency medicine, family medicine, internal medicine, and pediatrics), directors of urology residencies, medical student educators in urology, and applicants to the 2006 urology residency match were invited to complete an online survey. Participants were asked to select the 5 most important topics to be included in a core urology curriculum for all medical students. RESULTS: Among 1859 stakeholders surveyed, response rates by stakeholder cohort ranged from 67% to 26% (overall 44%). There was marked homogeneity among participant groups as to what respondents considered to be the most important topics for a core urology curriculum for medical students. Based on aggregate data, the 8 most commonly cited topics included urinary stone disease (75% of respondents), hematuria (65%), urinary tract infections in adults (53%), benign prostatic hyperplasia (52%), urinary incontinence (45%), prostate cancer (45%), screening with prostate-specific antigen (33%), and testis torsion (24%). CONCLUSIONS: This survey has identified the most important urology topics about which medical students should learn before graduation. Work is currently under way under the aegis of the American Urological Association to develop materials to standardize student education in these core topics across the United States. PMID- 18387384 TI - Do "rapid" PSA assays reduce anxiety and stress of prostate cancer patients undergoing regular review? A prospective evaluation. AB - OBJECTIVES: Waiting for prostate-specific antigen (PSA) results may create anxiety for patients. Recently developed "rapid" PSA assays have become available, which achieve laboratory sensitivity and specificity. The manufacturers claim these assays will help to reduce anxiety associated with PSA testing. The aim of this study was to test the hypothesis that rapid received PSA results reduce patient anxiety. METHODS: One hundred eighty-eight (n = 188) patients participated in a prospective randomized study. After obtaining informed consent, 67 patients were assigned to receive PSA results within 15 minutes of the blood sample being drawn, facilitating a discussion with the physician while in-clinic. One hundred twenty-one (n = 121) patients were assigned to receive their results within 1 to 4 days by telephone. Patients completed a baseline questionnaire about PSA testing and a follow-up questionnaire after they had received their PSA result by either method. RESULTS: There were no significant differences in patient demographics between the "rapid" and "delayed" PSA test groups. Baseline measurements of stress and anxiety were low and not significantly different between the groups. Receiving a rapid PSA result did not significantly reduce stress and anxiety compared with a delayed result. However, 89% of patients receiving a rapid result would elect to have this method again. CONCLUSIONS: The rapid PSA test did not prove to alleviate stress or anxiety associated with receiving results. However, the cohort assigned to the rapid PSA test would prefer to have their results rapidly to facilitate discussion regarding their future management. PMID- 18387385 TI - Staging saturation biopsy in patients with prostate cancer on active surveillance protocol. AB - OBJECTIVES: One option for the management of low-grade, low-stage prostate cancer is to delay or forego treatment unless evidence of an increased risk of disease progression exists. Accurate assessment of the disease extent and aggressiveness is necessary to determine the candidates for active surveillance (AS). Office based saturation prostate biopsy (SB) provides more accurate staging than traditional biopsy; therefore, we studied its role in patients on an AS protocol. METHODS: Our database identified 52 men with prostate cancer treated with AS from July 2000 to May 2007. The records were reviewed to determine the role of SB in determining the need for definitive therapy. RESULTS: The patients had a median age of 69 years (range 51 to 83) and median prostate-specific antigen (PSA) level of 5.1 ng/mL (range 0.5 to 47). Patients underwent subsequent staging 20-core SB a median of 9 months (range 1 to 20 months) after diagnosis. The disease of 20 patients (38%) was upstaged as defined by an increase in Gleason score or increased disease volume, leading to a recommendation for active treatment. Patients with disease upstaging had had significantly fewer cores taken at the initial diagnostic biopsy (11% with 20 cores or more compared with 55% with fewer than 20 cores, P = 0.002). CONCLUSIONS: SB might lead to a more accurate assessment of the extent and grade of disease in men with prostate cancer on an AS protocol than traditional biopsy. In our series, more than one half of patients who pursue an AS protocol delayed or avoided local therapy. No patient developed clinical metastasis, but long-term surveillance is required. PMID- 18387386 TI - Role of laparoscopic stone surgery. AB - OBJECTIVES: To define the role of laparoscopic stone surgery in current urologic practice. METHODS: All cases of laparoscopic surgery for renal calculus disease performed at a single institution during a 2-year period were reviewed. A total of 449 stone-removing procedures were performed by one of us (D.G.A.) during this interval, of which 5 were laparoscopic (1.1%). Of the five laparoscopic procedures, three were pyelolithotomy/pyeloplasty, one was laparoscopic-assisted percutaneous nephrostolithotomy, and one was caliceal diverticular stone removal. RESULTS: All 5 patients were rendered stone free, and no patient had perioperative complications. CONCLUSIONS: A laparoscopic approach is rarely needed in patients requiring stone removal. However, it can be a safe and effective approach in the management of select patients with complex stone disease. PMID- 18387387 TI - Shock wave lithotripsy for renal stones is not associated with hypertension and diabetes mellitus. AB - OBJECTIVES: To clarify the influences of shock wave lithotripsy (SWL) treatments for renal and ureteropelvic junction stones on new onsets of hypertension and diabetes mellitus (DM). METHODS: We compared the new onsets of hypertension and DM after SWL in renal and ureteral stone groups. The renal stone group consisted of 772 patients treated with SWL, who had a possibility of renal and pancreatic shock wave damage. The ureteral stone group consisted of 505 patients treated with SWL, who were unlikely to have suffered SWL damage in the kidney and pancreas areas. Both treatment groups received SWL between 1984 and 1994. RESULTS: The rates of new onset of hypertension in the renal stone and ureteral stone groups were 22.8% and 20.0% in men and 23.1% and 20.5% in women, respectively. The rates of new onset of DM in the renal stone and ureteral stone groups were 7.4% and 11.0% in men and 8.7% and 8.7% in women, respectively. There was no significant difference in the new onsets of hypertension and DM between renal and ureteral stone groups depending on each age decade or sex. Treatment for renal stone was not a significant risk factor for new onsets of hypertension and DM by logistic regression analysis. CONCLUSIONS: Our findings suggest that SWL treatment for renal and UPJ stones might not be associated with new onset of hypertension or DM. PMID- 18387389 TI - Trainees do not negatively impact the institutional learning curve for robotic prostatectomy as characterized by operative time, estimated blood loss, and positive surgical margin rate. AB - OBJECTIVES: We evaluated the learning curves and perioperative outcomes of an experienced laparoscopic surgeon and his trainees to assess our structured teaching program. METHODS: We retrieved 383 patients undergoing robot-assisted laparoscopic prostatectomy (RALP) from our database. Trainees completed a structured teaching program and were categorized as early (days 0 to 232), mid (days 566 to 797), and late (days 825 to 1218) according to the time period in which they were working with the mentor. We compared operative times, estimated blood loss (EBL), and positive surgical margin (PSM) rates between the trainees and the mentor (Mann-Whitney and Chi-square test). Association of EBL, body mass index (BMI), and prostate weight with operative time was evaluated in multivariate linear regression analysis. RESULTS: Median operative times of the early, mid, and late trainees (258, 220, and 200 minutes) significantly decreased and were similar to the corresponding senior surgeon's (254, 242, and 180 minutes). Operative times decreased with lower BMI, EBL, and prostate weight (P = 0.006, P <0.001, and P <0.001, respectively). Overall, EBL (150 mL vs. 150 mL, P = 0.215) and PSM rates (20% vs. 18.6%, P = 0.741) did not differ between the mentor and the trainees. CONCLUSIONS: A structured teaching program for RALP is effective and trainees are able to adopt the increased efficiency and skills of their mentor. Lower BMI, EBL, and prostate weight were associated with shorter operative times. Trainees performing the procedure did not negatively affect EBL and positive surgical margin rate. PMID- 18387391 TI - Urinary epidermal growth factor and interleukin-6 levels in patients with painful bladder syndrome/interstitial cystitis treated with cyclosporine or pentosan polysulfate sodium. AB - OBJECTIVES: To measure urinary epidermal growth factor (EGF) and interleukin-6 (IL-6) levels in painful bladder syndrome/interstitial cystitis (PBS/IC) patients randomized to either cyclosporine A (CyA) or pentosan polysulfate sodium (PPS) treatment for 6 months and compare the results with treatment response. METHODS: Urine samples from PBS/IC patients were collected before (n = 37) and after treatment (n = 34). The concentration of EGF was determined by an in-house immunofluorometric assay and that of IL-6 by the Immulite assay. The clinical response to treatment was determined by the subjective global response analysis (GRA, scale 1 to 6). Participants who reported categories 5 (much better) or 6 (cured) were considered responders. RESULTS: According to GRA, 72% of CyA patients and 16% of PPS patient responded to their treatment (P <0.001). In the CyA group, post-treatment urinary EGF levels were significantly reduced (from 35 +/- 15.8 to 28.3 +/- 17.9 ng/mg creatinine; P <0.034), whereas the urinary IL-6 levels were not affected by CyA or PPS treatment in the whole group. In older patients (aged more than 52 years) who had higher IL-6 levels and longer duration of disease, these were reduced by successful CyA therapy. CONCLUSIONS: Treatment of PBS/IC patients with CyA resulted in significantly decreased urinary levels of EGF. Interleukin-6 levels did not change significantly in all treated patients after either CyA or PPS treatment, but in older patients the levels were reduced after CyA treatment. PMID- 18387392 TI - Characterization of a clinical cohort of 87 women with interstitial cystitis/painful bladder syndrome. AB - OBJECTIVE: To provide a characterization of a cohort of women with interstitial cystitis/painful bladder syndrome (IC/PBS) by describing their historical and clinical characteristics. This was reported with the National Institutes of Health chronic prostatitis cohort, but a literature review did not reveal a similar study for women with IC/PBS. METHODS: A total of 87 women with IC/PBS were referred to the Beaumont Women's Initiative for Pelvic Pain and Sexual Health program. A certified nurse practitioner took a comprehensive history and performed a pelvic exam for each. Data were analyzed using descriptive statistics to describe this cohort. RESULTS: Most women experienced constant pain for 5 or more years (mean Visual Analog Scale = 5 out of 10). A total of 94.2% had levator pain. More than 50% had vulvar pain with exam. More than half reported a history of abuse, often in more than one life stage. A total of 28% had cesarean births and 76% had a history of miscarriage, stillbirth, or abortion. Women averaged 4 lifetime pelvic surgeries, and 48% had hysterectomies, two-thirds of which were done before IC/PBS diagnosis. Premenstrual women reported pain throughout the menstrual cycle. As many as 12% had chlamydia previously, which was higher than the national average. Common comorbidities were pelvic pain (93%), allergies (86%), and sexual dysfunction (72%). CONCLUSIONS: This population of women with unrelieved chronic pain, frequency, and urgency is in desperate need of care. Researchers should continue to search for the etiology, prevention, and treatment interventions that are effective in dealing with IC/PBS. It may be most therapeutic to develop a multimodal plan of care that includes physical therapy, oral and intravesical therapies, neuromodulation, and cognitive-behavioral therapies. PMID- 18387393 TI - Acute effect of posterior tibial nerve stimulation on neurogenic detrusor overactivity in patients with multiple sclerosis: urodynamic study. AB - OBJECTIVES: The aim of this study was to determine the acute urodynamic effects of posterior tibial nerve stimulation (PTNS) in patients with neurogenic detrusor overactivity (NDO) caused by multiple sclerosis (MS). METHODS: A total of 29 patients of whom there were 12 (41.3%) men and 17 (58.6%) women with NDO, mean age 46.5 +/- 8.5 years (range 29 to 55 years) enrolled in the study. Mean duration of MS was 8.80 +/- 3.6 years; duration of urinary disorder was 4.3 +/- 1.8 years. The expanded disability status scale (EDSS) of the study was detected 4.8 +/- 1.9. Urodynamic studies before and during PTNS were performed. Electrical stimulation was applied unilaterally from the medial malleolus and posterior to the edge of the tibia by using charge-compensated 200 microsecond pulses with a pulse rate of 20 Hz. Mean first involuntary detrusor contractions and mean maximum cystometric capacity before and during PTNS were compared. RESULTS: Mean first involuntary detrusor contraction on standard cystometry was 138.34 +/- 6.36 mL (60 to 225 mL), whereas it was 230.48 +/- 8.89 mL (145 to 375 mL) during PTNS. Mean maximum cystometric capacity on standard cystometry was 193.93 +/- 9.90 mL (110 to 304 mL), whereas it was 286.48 +/- 9.09 mL (221 to 376 mL) during stimulation. The improvements in the first involuntary detrusor contraction and maximum cystometric capacity were statistically significant during stimulation (P <0.001). The difference of mean first involuntary detrusor contraction volume and mean maximum cystometric capacity at baseline and after PTNS was statistically significant (P <0.001). CONCLUSIONS: These results have demonstrated the objective effect of acute PTNS on urodynamic parameters. PTNS is effective to suppress detrusor overactivity in MS patients. PMID- 18387394 TI - Late failures after laparoscopic pyeloplasty. AB - OBJECTIVES: We present the results of our series of 65 consecutive laparoscopic pyeloplasties, which include some unexpected late failures. METHODS: We retrospectively reviewed the perioperative course of 65 consecutive patients who underwent transperitoneal laparoscopic pyeloplasty from August 1996 through July 2005 at our institution. RESULTS: Of the obstructions, 47 and 18 were primary and secondary, respectively. Laparoscopic pyeloplasty was successfully performed on 64 patients. Of the patients, 42 (65%) had crossing vessel(s) found intraoperatively. We performed dismembered pyeloplasty for 50 (77%), Y-V plasty for 12 (18%) and Fengerplasty for 3 patients (5%). Mean laparoscopic operative time was 218 minutes, not including a mean of 40 minutes for stent placement and repositioning. The median hospital stay was 2 days. All 9 major complications (14%) pertained to urinary leakage/ureteral obstruction. Of the 60 patients with follow-up, 7 (12%) failed within 1 year of surgery. An additional 3 patients who appeared cured at initial follow-up presented with late recurrences, at 2, 2.5, and 6 years postoperatively. The overall success rate at 1 year was 88%, with a long-term success rate of 83%, and was not related to type of obstruction or repair, or urinary leakage/ureteral obstruction. CONCLUSIONS: Contrary to findings in other series, 30% of our failures occurred 2 or more years after pyeloplasty. All failures, however, were symptomatic, such that radiographic follow-up after 1 year in patients with symptoms preoperatively may not be necessary. PMID- 18387397 TI - Randomized, double-blind, crossover trial of sildenafil in men with mild to moderate erectile dysfunction: efficacy at 8 and 12 hours postdose. AB - OBJECTIVES: To clarify the period of responsiveness to sildenafil. METHODS: Under a double-blind protocol, men with mild to moderate erectile dysfunction (International Index of Erectile Function [IIEF] Erectile Function domain score, 11 to 25) were randomized to sildenafil (100 mg) or placebo and attempted intercourse 8 hours (range, 7 to 9 hours) postdose (first 4-week phase) and 12 hours (11 to 13 hours) postdose (second 4-week phase after treatment crossover). The primary outcome was the per-patient proportion (PPP; least squares means [95% confidence interval]) of affirmative responses to the Sexual Encounter Profile question 3 (SEP3: "Did your erection last long enough for you to have successful intercourse?"). RESULTS: For sildenafil (n = 174) versus placebo (n = 177), baseline values were similar but the PPP of successful intercourse attempts increased to 76% (69% to 82%) versus 50% (43% to 57%) in phase 1 (odds ratio [OR] = 3.2) and 79% (72% to 85%) versus 52% (44% to 60%) in phase 2 (OR = 3.5), and the PPP of Erection Hardness Score 4 erections (completely hard and fully rigid) was 41% (34% to 48%) versus 10% (7% to 15%) in phase 1 (OR = 6.2) and 44% (37% to 51%) versus 17% (12% to 23%) in phase 2 (OR = 4.0). Thus, at 12 hours, the odds of successful intercourse tripled and of a completely hard erection quadrupled. The sildenafil group achieved greater (P <0.001) PPP of successful penetration (SEP2), satisfaction with erection hardness (SEP4), and satisfaction with the sexual experience (SEP5); improvement in IIEF domain scores; and treatment satisfaction on the Erectile Dysfunction Inventory of Treatment Satisfaction. CONCLUSIONS: In men with mild to moderate ED, responsiveness to sildenafil may persist much longer than 4 hours. PMID- 18387398 TI - Subjective patient-reported experiences after surgery for Peyronie's disease: corporeal plication versus plaque incision with vein graft. AB - OBJECTIVES: To compare patient-perceived outcomes of corporeal plication to plaque incision with saphenous vein grafting for the correction of Peyronie's disease. METHODS: Patients with stable Peyronie's disease deemed to be good operative candidates for both tunical plication and plaque incision with saphenous vein graft were counseled on both procedures and chose which operation they would undergo. At 1 year, the records were reviewed and the patients were contacted. The variables included age, operative time, and outcome ("satisfactorily straight," loss of rigidity, loss of sensation, new use of erectile aids, ability to have intercourse, palpable nodules, erectile pain, penile shortening, and being "completely satisfied"). RESULTS: Of the 67 patients, 35 underwent tunical plication and 32 underwent plaque incision with vein grafting. No differences were present in patient age between the two groups. The average operative time was shorter for the plication group (P = 0.0001). No differences were found regarding satisfactory straightness (P = 0.13), satisfaction with the operation (P = 0.71), new use of erectile aids (P = 0.06), erectile pain (P = 0.12), or subjective penile shortening (P = 0.41). Patients who underwent plaque incision with grafting were more likely to experience loss of rigidity (P = 0.03), inability to have intercourse (P = 0.05), and loss of sensation (P = 0.0045). Patients who underwent plication were more likely to experience palpable nodules (P = 0.03). CONCLUSIONS: The results of our study have shown that both procedures are effective surgical options for the correction of Peyronie's disease. Plication is a simple procedure with less morbidity. Shortening is a common complaint, regardless of the type of operation done. PMID- 18387399 TI - Effects of statin treatment on serum sex steroids levels and autonomic and erectile function. AB - OBJECTIVES: To investigate the effect of statin treatment on serum sex steroid levels, heart rate variability, erectile function, and libido in patients with hyperlipidemia. METHODS: A total of 74 patients (mean age 44.7 +/- 7.1 years) with hyperlipidemia were enrolled into this study. After a cardiac examination, the serum lipid levels were measured, and the 24-hour Holter monitoring, heart rate variability, and autonomic test results were also evaluated. Erectile function was assessed using the International Index of Erectile Function (IIEF) questionnaire. Later, atorvastatin 40 mg/day was initiated in all patients and used for the subsequent 12 months. All diagnostic tests (cardiac, biochemical, and autonomic and the IIEF questionnaire) were performed again at 6 and 12 months of follow-up. RESULTS: A statistically significant decrease was found in the serum lipid levels at 6 months (P <0.05). In contrast, the average IIEF scores (24.7 +/- 6.4 at baseline) had increased to 25.0 +/- 4.9 and 26.1 +/- 5.9 at 6 and 12 months of follow-up, respectively. Although the parasympathetic activities increased and sympathetic activities decreased with atorvastatin treatment, these changes were not statistically significant (P >0.05). In paired comparison, significant differences were found among the IIEF scores of the three periods (P = 0.013). The difference was more evident after 6 months of treatment (IIEF(1vs2) = 0.475; IIEF(1vs3) = 0.027; IIEF(2vs3) = 0.012). CONCLUSIONS: Although improvement in the lipid profile occurred early during the statin treatment, restoration of erectile function appeared later, which could be attributed to the restoration of endothelial functions by lowered serum lipid levels. PMID- 18387400 TI - Urinary cytology and quantitative BTA and UBC tests in surveillance of patients with pTapT1 bladder urothelial carcinoma. AB - OBJECTIVES: To compare results of urinary cytology, quantitative detection of human complement factor H-related protein (BTA TRAK), and urinary fragments of cytokeratins 8 and 18 (UBC IRMA) with the recurrence status in patients with pTapT1 bladder cancer and to define the possible role of these methods in a surveillance protocol. METHODS: We collected urine from 88 consecutive patients with primary pTapT1 tumors before the first transurethral resection (TURB) and before each follow-up cystoscopy. In all samples urinary cytology and quantitative BTA and UBC tests were performed. We compared results with recurrence status and with tumor characteristics in the case of recurrence. We constructed receiver operator characteristic (ROC) curves for quantitative methods. In addition, we evaluated individual cutoffs based on pretreatment levels. RESULTS: During the mean follow-up of 16.96 months, we performed 313 cystoscopies, 93 of which were positive in 51 patients. The sensitivity and specificity of cytology, BTA, and UBC were 19.8% and 99%, 53.8% and 83.9%, and 12.1% and 97.2%, respectively. The sensitivity of pTis detection was 66.6%, 0%, and 100%, respectively. With cutoffs set to a sensitivity of 90%, the specificity of BTA and UBC dropped to 24.8% and 20.4%, respectively. Individually calculated cutoffs did not provide a significant benefit. CONCLUSIONS: Because of high specificity and sensitivity in pTis detection, urinary cytology fulfills requirements for an adjunctive method to cystoscopy. Quantitative BTA and UBC tests have a low sensitivity in the detection of bladder cancer recurrence and cannot be used routinely to reduce the number of cystoscopies during follow-up. PMID- 18387401 TI - Relationship of biochemical outcome to percentage of positive biopsies in men with clinically localized prostate cancer treated with permanent interstitial brachytherapy. AB - OBJECTIVES: Recent studies have demonstrated that the percentage of positive prostate needle biopsies (PPBs) is an independent predictor of biochemical failure (BF) after radical prostatectomy and external beam radiotherapy. Few studies have confirmed this phenomenon in patients treated with permanent interstitial brachytherapy. We conducted an analysis to determine the clinical utility of the PPBs in predicting BF after permanent interstitial brachytherapy for patients with clinically localized prostate cancer. METHODS: The clinical and pathologic data from 245 hormone-naive patients with clinical Stage T1c-T2cNxM0 prostate cancer who had undergone permanent prostate brachytherapy from 1992 to 2002 at our institution were retrospectively evaluated. The PPBs, preimplant prostate-specific antigen, and biopsy Gleason score were evaluated as predictors of BF after brachytherapy. The risk of BF was calculated using logistic regression models. The time to BF was assessed using Cox proportional hazard models and Kaplan-Meier estimates. RESULTS: The median PPBs in our study cohort was 27%. The risk of BF was 10.9% for those with a PPBs less than the median versus 26.2% for those with a PPBs greater than the median. A PPBs greater than the median was an independent predictor of BF on multivariate analysis (relative risk 2.3, 95% confidence interval 1.21 to 4.39, P = 0.011). In the time-to failure models, the PPBs, prostate-specific antigen level, and Gleason score were significantly associated with BF. CONCLUSIONS: Our data suggest that the PPBs is significantly associated with BF in patients undergoing permanent interstitial brachytherapy. The PPBs, along with the pretreatment prostate-specific antigen level and Gleason biopsy score, is a useful preoperative parameter in predicting for BF and the time to failure for patients treated with interstitial brachytherapy. PMID- 18387402 TI - Renal hemangiopericytoma: case report and review of the literature. AB - Hemangiopericytoma is an uncommon perivascular tumor that occurs most frequently in the pelvis, head and neck, and meninges; it is extremely rare in the kidney. We report a kidney hemangiopericytoma in a 43-year-old woman who was treated with a nephron-sparing surgery and review the literature. PMID- 18387403 TI - Re: Desai MR, Ganpule AP, Gupta R, et al: Outcome of renal transplantation with multiple versus single renal arteries after laparoscopic live donor nephrectomy: a comparative study (Urology 69: 824-827, 2007). PMID- 18387405 TI - Multivitamin supplements and cataract prevention. PMID- 18387406 TI - A randomized, double-masked, placebo-controlled clinical trial of multivitamin supplementation for age-related lens opacities. Clinical trial of nutritional supplements and age-related cataract report no. 3. AB - OBJECTIVE: To evaluate the effect of a multivitamin/mineral supplement on development or progression of age-related lens opacities. DESIGN: Randomized, double-masked, single center, placebo-controlled clinical trial. PARTICIPANTS: One thousand twenty participants, 55 to 75 years old and with early or no cataract, were randomly assigned to a daily tablet of a multivitamin/mineral formulation or a placebo. METHODS: Baseline and annual lens photographs were graded for severity of lens opacities according to a modification of the Age Related Eye Disease Study system for classifying cataracts. MAIN OUTCOME MEASURES: The primary outcome was a prespecified increase from baseline in nuclear, cortical, or posterior subcapsular cataract (PSC) opacity grades or cataract surgery. Secondary outcomes included an increase in type-specific opacity grades, cataract surgery, and visual acuity (VA) loss from baseline > or =15 letters. RESULTS: Participants were observed for an average of 9.0+/-2.4 years. There was a decrease in total lens events in participants assigned to the multivitamin/mineral formulation compared with those assigned to the placebo (hazard ratio [HR], 0.82; 95% confidence interval [CI], 0.68-0.98; P = 0.03). Nuclear events were significantly less common (HR, 0.66; 95% CI, 0.50-0.88; P = 0.004) and PSC events significantly more common (HR, 2.00; 95% CI, 1.35-2.98; P<0.001) in participants taking the multivitamin/mineral formulation than in those assigned to the placebo. No statistically significant treatment effects were seen for cortical opacities, moderate VA loss, or cataract surgery. CONCLUSIONS: Lens events were less common in participants who took the multivitamin/mineral formulation, but treatment had opposite effects on the development or progression of nuclear and PSC opacities, the 2 most visually important opacity subtypes. PMID- 18387410 TI - Intravenous immunoglobulins and mucous membrane pemphigoid. PMID- 18387409 TI - Correlation of ophthalmic examination with carrier status in females potentially harboring a severe Norrie disease gene mutation. AB - PURPOSE: To correlate ophthalmic findings with carrier status for a severe Norrie disease (ND) gene mutation (C95F). DESIGN: Prospective interventional case series. PARTICIPANTS: Six potential carriers and 1 obligate carrier from a family harboring the mutation. METHODS: An ophthalmologist blind to the pedigree performed a full ophthalmic examination for the 7 asymptomatic family members. A peripheral blood sample was collected from each for ND gene sequencing. MAIN OUTCOME MEASURES: Ophthalmic examination findings (with attention to the presence or absence of retinal findings) and results of ND gene sequencing. RESULTS: Three carriers were identified by molecular genetics, and all 3 of them had peripheral retinal abnormality. However, 3 of the 4 genetically identified noncarriers also exhibited peripheral retinal abnormality. Two of these noncarriers with retinal findings were the offspring of a confirmed noncarrier. The genetically identified noncarrier with a normal peripheral retinal examination was the daughter of an obligate carrier. CONCLUSIONS: The presence of peripheral retinal changes was not useful for carrier prediction in a family harboring ND. There are likely additional loci responsible for phenotypic expression. PMID- 18387411 TI - Anesthesia for phacotrabeculectomy. PMID- 18387407 TI - The effect of donor age on corneal transplantation outcome results of the cornea donor study. AB - OBJECTIVE: To determine whether graft survival over a 5-year follow-up period using corneal tissue from donors older than 65 is similar to graft survival using corneas from younger donors. DESIGN: Multicenter prospective, double-masked, controlled clinical trial. PARTICIPANTS: One thousand ninety subjects undergoing corneal transplantation for a moderate-risk condition (principally Fuchs' dystrophy or pseudophakic corneal edema); 11 subjects with ineligible diagnoses were not included. METHODS: Forty-three participating eye banks provided corneas from donors in the age range of 12 to 75 with endothelial cell densities of 2300 to 3300 cells/mm(2), using a random approach without respect to recipient factors. The 105 participating surgeons at 80 sites were masked to information about the donor cornea including donor age. Surgery and postoperative care were performed according to the surgeons' usual routines. Subjects were observed for 5 years. MAIN OUTCOME MEASURES: Graft failure, defined as a regraft or a cloudy cornea that was sufficiently opaque as to compromise vision for a minimum of 3 consecutive months. RESULTS: The 5-year cumulative probability of graft survival was 86% in both the <66.0 donor age group and the >/=66.0 donor age group (difference = 0%, upper limit of 1-sided 95% confidence interval = 4%). In a statistical model with donor age as a continuous variable, there was no significant relationship between donor age and outcome (P = 0.11). Three graft failures were due to primary donor failure, 8 to uncorrectable refractive error, 48 to graft rejection, 46 to endothelial decompensation (23 of which had a prior, resolved episode of probable or definite graft rejection), and 30 to other causes. Distributions of the causes of graft failure did not differ between donor age groups. CONCLUSIONS: Five-year graft survivals for cornea transplants at moderate risk for failure are similar using corneas from donors >/= 66.0 years and donors < 66.0. Surgeons and patients now have evidence that corneas comparable in quality to those used in this study from donors through age 75 are suitable for transplantation. PMID- 18387412 TI - Artisan phakic intraocular lens. PMID- 18387408 TI - Donor age and corneal endothelial cell loss 5 years after successful corneal transplantation. Specular microscopy ancillary study results. AB - OBJECTIVE: To determine whether endothelial cell loss 5 years after successful corneal transplantation is related to the age of the donor. DESIGN: Multicenter, prospective, double-masked clinical trial. PARTICIPANTS: Three hundred forty seven subjects participating in the Cornea Donor Study who had not experienced graft failure 5 years after corneal transplantation for a moderate-risk condition (principally Fuchs' dystrophy or pseudophakic corneal edema). TESTING: Specular microscopic images of donor corneas obtained before surgery and postoperatively at 6 months, 12 months, and then annually through 5 years were submitted to a central reading center to measure endothelial cell density (ECD). MAIN OUTCOME MEASURE: Endothelial cell density at 5 years. RESULTS: At 5 years, there was a substantial decrease in ECD from baseline for all donor ages. Subjects who received a cornea from a donor 12 to 65 years old experienced a median cell loss of 69% in the study eye, resulting in a 5-year median ECD of 824 cells/mm(2) (interquartile range, 613-1342), whereas subjects who received a cornea from a donor 66 to 75 years old experienced a cell loss of 75%, resulting in a median 5 year ECD of 654 cells/mm(2) (interquartile range, 538-986) (P [adjusted for baseline ECD] = 0.04). Statistically, there was a weak negative association between ECD and donor age analyzed as a continuous variable (r [adjusted for baseline ECD] = -0.19; 95% confidence interval, -0.29 to -0.08). CONCLUSIONS: Endothelial cell loss is substantial in the 5 years after corneal transplantation. There is a slight association between cell loss and donor age. This finding emphasizes the importance of longer-term follow-up of this cohort to determine if this relationship affects graft survival. PMID- 18387414 TI - Refraction after IOL exchange. PMID- 18387416 TI - Talc retinopathy. PMID- 18387418 TI - Foveal depression and albinism. PMID- 18387420 TI - Latitude and uveal melanoma. PMID- 18387421 TI - Lacrimal surgery. PMID- 18387423 TI - [Risk of infection and gastrointestinal endoscopy]. PMID- 18387424 TI - [An uncommon use of argon plasma coagulation: endoscopic transection of self expanding metallic stents]. AB - Migration, distal malposition or fracture of self-expanding metallic biliary or esophageal stents rarely occur but may lead to stent dysfunction, mucosal trauma secondary to impaction or friction on the digestive wall. We report our experience of using argon plasma to endoscopically cut three metallic biliary and esophageal self-expanding stents to restore permeability or to treat impaction of the stent after migration, malposition or fracture. PMID- 18387425 TI - [Splenic trauma: an unusual complication of colonoscopy with polypectomy]. AB - Nowadays, colonoscopy has become an invaluable tool in the diagnosis and treatment of diseases of the colon and rectum. Colonoscopy is still an invasive exam with several complications. The most common complications are perforation and bleeding, which occur in up to 1% after diagnostic colonoscopy and 3% of patients undergoing therapeutic colonoscopy. Less common complications include pneumothorax, pneumomediastinum, colonic volvulus, hernia incarceration, retroperitoneal abscess and mesenteric tear. Splenic rupture is a rare and potentially lethal colonoscopic complication with less than 45 cases reported in the world. The overall incidence is 0.004%. Mechanisms of injury and available treatment options remain discussed. We present a case of splenic rupture after colonoscopy with polypectomy in a 73-year-old woman managed first with nonoperative treatment and nine days later with surgical treatment. As the indications for colonoscopy expand, including the introduction of mass screening for colorectal cancer, endoscopists should be increasingly aware of that life threatening complication after colonoscopy and know the emergency treatment. PMID- 18387426 TI - A double blind randomized controlled trial of a probiotic combination in 100 patients with irritable bowel syndrome. AB - OBJECTIVES: The purpose of this study was to evaluate the effects of a probiotic combination on symptoms in patients with irritable bowel syndrome (IBS). METHODS: We investigated the efficiency of a probiotic dietary supplement, containing four strains of lactic acid bacteria, on symptoms of IBS. One hundred and sixteen patients with IBS fulfilling the Rome II criteria were randomized in a parallel group, double-blind study to receive a placebo or a probiotic combination (1 x 10(10) cfu once daily) for four weeks. The symptoms that were monitored weekly included discomfort, abdominal pain, and stool frequency and quality. Quality of life was assessed before and at the end of the treatment using the SF36 and FDD quality-of-life questionnaires. RESULTS: One hundred subjects completed the study (48 probiotic combination, 52 placebo). The probiotic combination was not superior to the placebo in relieving symptoms of IBS (42.6 versus 42.3% improvement). However, the decrease of abdominal pain between the first and the fourth week of treatment was significantly higher in probiotic treated patients ( 41.9 versus -24.2%, P=0.048). Interesting findings from the IBS sub-groups were also observed such as a lower pain score at end point in patients with alternating bowel habits (P=0.023) and an increase of stool frequency in the constipated sub-group from the first week of probiotic treatment (P=0.043). CONCLUSIONS: The probiotic combination was not significantly superior to the placebo in relieving symptoms of IBS. Despite the apparent high placebo response, interesting findings from IBS sub-groups were observed in the field of abdominal pain and stool frequency. PMID- 18387427 TI - [Retrobulbar optic neuritis during infliximab treatment for ulcerative colitis]. PMID- 18387428 TI - [A rare complication of immunosuppression: digestive aspergillosis]. PMID- 18387429 TI - [Ectopic sebaceous glands in the esophagus]. AB - Ectopic sebaceous glands have been detected in many tissues of ectodermal origin, but their presence in the esophagus remain, until now, a very rare anomaly. We report two new cases diagnosed by endoscopic biopsy. PMID- 18387430 TI - [Endocrine tumor arising in heterotopic gastric pancreas]. AB - We report the case of a 49-year-old caucasian woman, in whom an endocrine tumor arising in gastric heterotopic pancreas was diagnosed. The patient was treated surgically with a gastric wedge resection. Heterotopic pancreas is a benign anatomic condition, probably widely underdiagnosed because usually asymptomatic. The malignant transformation of aberrant pancreas is very rare and almost always in adenocarcinoma. The endocrine tumors developed in heterotopic pancreas are exceedingly rare. Of our knowledge, only four cases have been published and only one case in the gastric location similar to this reported case. PMID- 18387431 TI - [Ectopic pancreas: an exceptional cause of lower intestinal bleeding with hemorrhagic shock]. PMID- 18387432 TI - Genomics: the next step to elucidate the etiology of calcific aortic valve stenosis. AB - With the current shift toward an older population, calcific aortic valve stenosis (AVS) is likely to become a major societal and economic burden. For many years, AVS was regarded as a degenerative and nonmodifiable process. However, molecular studies unanimously demonstrated that AVS is an actively regulated disorder with several potential therapeutic targets. Many factors are predicted to cause AVS, and an important genetic predisposition is anticipated. In this review, we describe candidate genes and signaling pathways identified by genetic research and incorporate this new knowledge into a more comprehensive picture of factors involved in the pathogenesis of AVS. We also emphasize the need for additional studies to elucidate its complete genetic architecture. Recent advances in genomic research offer a remarkable opportunity to investigate AVS at the most fundamental level. The benefits of these new approaches can be observed in many complex diseases, but the field of AVS is trailing behind. We discuss the future utility of these new genomic approaches to improve our understanding of AVS and to refine the management of patients in terms of diagnosis, prevention, and treatment. PMID- 18387433 TI - Community-based provision of statin and aspirin after the detection of coronary artery calcium within a community-based screening cohort. AB - OBJECTIVES: We examined the association of coronary artery calcium (CAC) detected on a screening exam with subsequent statin and aspirin usage in a healthy male screening cohort. BACKGROUND: Whether the presence of CAC, an independent predictor of coronary heart disease outcomes, alters clinical management, such as the use of preventive medications, is unknown. METHODS: Men (n = 1,640) ages 40 to 50 years (mean 42 years) were screened for coronary heart disease risk factors and CAC. The CAC scores and risk factors were reported to patients, and results were made available in the electronic medical record; however, medications were not prescribed or recommended by the study. During up to 6 years of subsequent annual structured telephone follow-up, we observed the community-based initiation and persistence of aspirin and statin therapy. RESULTS: A progressive increase in the incidence of pharmacotherapy was noted over time such that those with CAC were 3 times more likely to receive a statin (48.5% vs. 15.5%, p < 0.001) and also significantly more likely to receive aspirin (53.0% vs. 32.3%; p < 0.001) than those without CAC. In multivariable models controlling for National Cholesterol Education Program risk variables and baseline medication use, CAC was strongly and independently associated with use of either statin (odds ratio [OR] 3.53; 95% confidence interval [CI] 2.66 to 4.69), aspirin (OR 3.05; 95% CI 2.30 to 4.05) or both (OR 6.97; 95% CI 4.81 to 10.10). CONCLUSIONS: In this prospective cohort, the presence of coronary calcification was associated with an independent 3-fold greater likelihood of statin and aspirin usage. PMID- 18387434 TI - Reversibility of cardiac abnormalities in morbidly obese adolescents. AB - OBJECTIVES: The purpose of this study was to evaluate changes in cardiac geometry, systolic and diastolic function before and after weight loss in morbidly obese adolescents. BACKGROUND: Cardiac abnormalities are present in morbidly obese adolescents; however, it is unclear if they are reversible with weight loss. METHODS: Data from 38 adolescents (13 to 19 years; 29 females, 9 males, 33 Caucasians, 5 African Americans) were evaluated before and after bariatric surgery. Left ventricular mass (LVM), left ventricular (LV) geometry, systolic and diastolic function were assessed by echocardiography. Mean follow up was 10 +/- 3 months. RESULTS: Weight and body mass index decreased post operatively (mean weight loss 59 +/- 15 kg, pre-operative body mass index 60 +/- 9 kg/m(2) vs. follow-up 40 +/- 8 kg/m(2), p < 0.0001). Change in LVM index (54 +/ 13 g/m(2.7) to 42 +/- 10 g/m(2.7), p < 0.0001) correlated with weight loss (r = 0.41, p = 0.01). Prevalence of concentric left ventricular hypertrophy (LVH) improved from 28% at pre-operative to only 3% at follow up (p = 0.007), and normal LV geometry improved from 36% to 79% at follow up (p = 0.009). Diastolic function also improved (mitral E/Ea lateral 7.7 +/- 2.3 at pre-operative vs. 6.3 +/- 1.6 at post-operative, p = 0.003). In addition, rate-pressure product improved suggesting decreased cardiac workload (p < 0.001). CONCLUSIONS: Elevated LVM index, concentric LVH, altered diastolic function, and cardiac workload significantly improve following surgically induced weight loss in morbidly obese adolescents. Large weight loss due to bariatric surgery improves predictors of future cardiovascular morbidity in these young people. PMID- 18387435 TI - Reduced nicotinamide adenine dinucleotide phosphate oxidase-derived superoxide and vascular endothelial dysfunction in human heart failure. AB - OBJECTIVES: We investigated the role of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in endothelial dysfunction in human heart failure. BACKGROUND: Vascular endothelial dysfunction in human heart failure contributes to increased tone, exercise limitation, and dysregulation of venous capacitance and vascular volume. The NADPH oxidases (Nox) are an important source of oxidative stress, but their role in the endothelial dysfunction of human heart failure remains unknown. METHODS: Endothelium-dependent and -independent vasorelaxation were assessed in saphenous vein segments obtained from consecutive patients with heart failure (n = 19) or normal left ventricular function (control; n = 35) undergoing coronary artery bypass graft. Saphenous vein superoxide production was measured by lucigenin-enhanced chemiluminescence and messenger ribonucleic acid expression of relevant transcripts quantified by real time polymerase chain reaction. RESULTS: Heart failure patients had significantly worse endothelial function than control subjects (15.2 +/- 3% vs. 40.5 +/- 8.4% relative relaxation; p < 0.05), elevated C-reactive protein (CRP) levels (8.6 +/- 2.7 mg/l vs. 2.6 +/- 0.4 mg/l; p < 0.05), over 2-fold higher NADPH-dependent superoxide generation (p < 0.05), and significantly higher expression of the Nox4 isoform and regulatory subunit p67phox. Superoxide levels were positively correlated with New York Heart Association functional class (r = 0.684; p < 0.05) and CRP (r = 0.501; p < 0.005; n = 32). CONCLUSIONS: Venous endothelial dysfunction in human heart failure is associated with increased Nox-derived superoxide generation. Inflammatory mechanisms may be involved in the increased reactive oxygen species generation. PMID- 18387436 TI - Inappropriate implantable cardioverter-defibrillator shocks in MADIT II: frequency, mechanisms, predictors, and survival impact. AB - OBJECTIVES: This study sought to identify the incidence and outcome related to inappropriate implantable cardioverter-defibrillator (ICD) shocks, that is, those for nonventricular arrhythmias. BACKGROUND: The MADIT (Multicenter Automatic Defibrillator Implantation Trial) II showed that prophylactic ICD implantation improves survival in post-myocardial infarction patients with reduced ejection fraction. Inappropriate ICD shocks are common adverse consequences that may impair quality of life. METHODS: Stored ICD electrograms from all shock episodes were adjudicated centrally. An inappropriate shock episode was defined as an episode during which 1 or more inappropriate shocks occurred; another inappropriate ICD episode occurring within 5 min was not counted. Programmed parameters for patients with and without inappropriate shocks were compared. RESULTS: One or more inappropriate shocks occurred in 83 (11.5%) of the 719 MADIT II ICD patients. Inappropriate shock episodes constituted 184 of the 590 total shock episodes (31.2%). Smoking, prior atrial fibrillation, diastolic hypertension, and antecedent appropriate shock predicted inappropriate shock occurrence. Atrial fibrillation was the most common trigger for inappropriate shock (44%), followed by supraventricular tachycardia (36%), and then abnormal sensing (20%). The stability detection algorithm was programmed less frequently in patients receiving inappropriate shocks (17% vs. 36%, p = 0.030), whereas other programming parameters did not differ significantly from those without inappropriate shocks. Importantly, patients with inappropriate shocks had a greater likelihood of all-cause mortality in follow-up (hazard ratio 2.29, p = 0.025). CONCLUSIONS: Inappropriate ICD shocks occurred commonly in the MADIT II study, and were associated with increased risk of all-cause mortality. PMID- 18387437 TI - Implantable cardioverter-defibrillator shocks: a double-edged sword? PMID- 18387438 TI - Occurrence and frequency of arrhythmias in hypertrophic cardiomyopathy in relation to delayed enhancement on cardiovascular magnetic resonance. AB - OBJECTIVES: Our aim was to determine whether myocardial fibrosis, detected by cardiovascular magnetic resonance (CMR), represents an arrhythmogenic substrate in hypertrophic cardiomyopathy (HCM). BACKGROUND: Myocardial fibrosis is identified frequently in HCM; however, the clinical significance of this finding is uncertain. METHODS: We studied prevalence and frequency of tachyarrhythmias on 24-h ambulatory Holter electrocardiogram (ECG) with regard to delayed enhancement (DE) on contrast-enhanced CMR in 177 HCM patients (age 41 +/- 16 yrs; 95% asymptomatic or mildly symptomatic). RESULTS: Premature ventricular contractions (PVCs), couplets, and nonsustained ventricular tachycardia (NSVT) were more common in patients with DE than those without DE (PVCs: 89% vs. 72%; couplets: 40% vs. 17%; NSVT: 28% vs. 4%; p < 0.0001 to 0.007). Patients with DE also had greater numbers of PVCs (202 +/- 655 vs. 116 +/- 435), couplets (1.9 +/- 5 vs. 1.2 +/- 10), and NSVT runs (0.4 +/- 0.8 vs. 0.06 +/- 0.4) than non-DE patients (all p < 0.0001); DE was an independent predictor of NSVT (relative risk 7.3, 95% confidence interval 2.6 to 20.4; p < 0.0001). However, extent (%) of DE was similar in patients with and without PVCs (8.2% vs. 9.1%; p = 0.93), couplets (8.5% vs. 8.4%; p = 0.99), or NSVT (8.3% vs. 8.5%; p = 0.35). CONCLUSIONS: In this large HCM cohort with no or only mild symptoms, myocardial fibrosis detected by CMR was associated with greater likelihood and increased frequency of ventricular tachyarrhythmias (including NSVT) on ambulatory Holter ECG. Therefore, contrast-enhanced CMR identifies HCM patients with increased susceptibility to ventricular tachyarrhythmias. PMID- 18387439 TI - Cardiovascular magnetic resonance for risk stratification of arrhythmia in hypertrophic cardiomyopathy. PMID- 18387440 TI - Pulse wave velocity is an independent predictor of the longitudinal increase in systolic blood pressure and of incident hypertension in the Baltimore Longitudinal Study of Aging. AB - OBJECTIVES: This study sought to evaluate whether pulse wave velocity (PWV), a noninvasive index of arterial stiffness, is a predictor of the longitudinal changes in systolic blood pressure (SBP) and of incident hypertension. BACKGROUND: Although arterial stiffness is believed to underlie, in part, the age associated changes in SBP, particularly at older ages, few longitudinal studies in humans have examined the relationship between arterial stiffness and blood pressure. METHODS: Pulse wave velocity was measured at baseline in 449 normotensive or untreated hypertensive volunteers (age 53 +/- 17 years). Repeated measurements of blood pressure were performed during an average follow-up of 4.9 +/- 2.5 years. RESULTS: After adjusting for covariates including age, body mass index, and mean arterial pressure, linear mixed effects regression models showed that PWV was an independent determinant of the longitudinal increase in SBP (p = 0.003 for the interaction term with time). In a subset of 306 subjects who were normotensive at baseline, hypertension developed in 105 (34%) during a median follow-up of 4.3 years (range 2 to 12 years). By stepwise Cox proportional hazards models, PWV was an independent predictor of incident hypertension (hazard ratio 1.10 per 1 m/s increase in PWV, 95% confidence interval 1.00 to 1.30, p = 0.03) in individuals with a follow-up duration greater than the median. CONCLUSIONS: Pulse wave velocity is an independent predictor of the longitudinal increase in SBP and of incident hypertension. This suggests that PWV could help identify normotensive individuals who should be targeted for the implementation of interventions aimed at preventing or delaying the progression of subclinical arterial stiffening and the onset of hypertension. PMID- 18387441 TI - Aging-related defects are associated with adverse cardiac remodeling in a mouse model of reperfused myocardial infarction. AB - OBJECTIVES: The purpose of this study was to study aging-associated alterations in the inflammatory and reparative response after myocardial infarction (MI) and their involvement in adverse post-infarction remodeling of the senescent heart. BACKGROUND: Advanced age is a predictor of death and ventricular dilation in patients with MI; however, the cellular mechanisms responsible for increased remodeling of the infarcted senescent heart remain poorly understood. METHODS: Histomorphometric, molecular, and echocardiographic end points were compared between young and senescent mice undergoing reperfused infarction protocols. The response of young and senescent mouse cardiac fibroblasts to transforming growth factor (TGF)-beta stimulation was examined. RESULTS: Senescence was associated with decreased and delayed neutrophil and macrophage infiltration, markedly reduced cytokine and chemokine expression in the infarcted myocardium, and impaired phagocytosis of dead cardiomyocytes. Reduced inflammation in senescent mouse infarcts was followed by decreased myofibroblast density and markedly diminished collagen deposition in the scar. The healing defects in senescent animals were associated with enhanced dilative and hypertrophic remodeling and worse systolic dysfunction. Fibroblasts isolated from senescent mouse hearts showed a blunted response to TGF-beta1. CONCLUSIONS: Although young mice exhibit a robust post-infarction inflammatory response and form dense collagenous scars, senescent mice show suppressed inflammation, delayed granulation tissue formation, and markedly reduced collagen deposition. These defects might contribute to adverse remodeling. These observations suggest that caution is necessary when attempting to therapeutically target the post-infarction inflammatory response in patients with reperfused MI. The injurious potential of inflammatory mediators might have been overstated, owing to extrapolation of experimental findings from young animals to older human patients. PMID- 18387442 TI - Aging mouse hearts are refractory to infarct size reduction with post conditioning. AB - OBJECTIVES: Our aim was to establish whether the efficacy of post-conditioning is maintained in aging hearts. BACKGROUND: Post-conditioning, or relief of myocardial ischemia in a stuttered manner, has been shown to reduce infarct size, in part because of up-regulation of survival kinases (extracellular-signal regulated kinase [ERK] 1/2 or PI3-kinase/Akt) during the early min of reperfusion. All of these data have, however, been obtained in adult populations; the question of whether post-conditioning-induced cardioprotection is maintained in aging cohorts is unknown. METHODS: Isolated buffer-perfused hearts were obtained from 3- to 4-month-old (adult) and 20- to 24-month-old C57BL/6J mice and subjected to 30 min of ischemia. For each cohort, hearts were randomized to receive standard, abrupt (control) reperfusion, or were post-conditioned with 3 or 6 10-s cycles of stuttered reflow. Primary end points were infarct size, cardiac expression of phospho-Akt, phospho-mitogen-activated protein kinase kinase 1/2 and phospho-ERK 1/2, and expression of mitogen-activated protein kinase-phosphatase-1 (MKP-1: phosphatase purported to play a primary role in ERK dephosphorylation). RESULTS: In adult mouse hearts, post-conditioning significantly reduced infarct size via up-regulation of ERK (but not Akt) signaling. In contrast, in the 2-year-old cohort, post-conditioning failed to limit necrosis, possibly a consequence of the deficit in ERK phosphorylation and increased MKP-1 expression seen in old hearts. Indeed, infusion of sodium orthovanadate, a nonspecific MKP inhibitor, attenuated MKP-1 expression and restored the post-conditioned phenotype in old hearts. CONCLUSIONS: Old mouse hearts are refractory to infarct size reduction with post-conditioning, possibly because of an age-associated increase in MKP-1 and resultant deficit in ERK phosphorylation. PMID- 18387443 TI - Aging and defective healing, adverse remodeling, and blunted post-conditioning in the reperfused wounded heart. PMID- 18387444 TI - Adjusted clopidogrel loading doses according to vasodilator-stimulated phosphoprotein phosphorylation index decrease rate of major adverse cardiovascular events in patients with clopidogrel resistance: a multicenter randomized prospective study. AB - OBJECTIVES: This study evaluates the clinical impact of adjusting the loading dose of clopidogrel according to vasodilator-stimulated phosphoprotein (VASP) index in patients with clopidogrel resistance undergoing percutaneous coronary intervention (PCI). BACKGROUND: Clopidogrel resistance plays a key role in ischemic recurrence after PCI. In vitro tests of clopidogrel resistance can accurately predict major adverse cardiac events after PCI. METHODS: In this prospective, randomized, multicenter study, clopidogrel resistance was defined as a VASP index of more than 50% after a 600-mg loading dose. Patients with clopidogrel resistance undergoing coronary stenting were randomized to a control group or to the VASP-guided group, in which patients received additional bolus clopidogrel to decrease the VASP index below 50%. RESULTS: A total of 162 patients were included. The control (n = 84) and VASP-guided groups (n = 78) had similar demographic, clinical, and biological characteristics. In the VASP-guided group, dose adjustment was efficient in 67 patients (86%) and VASP index was significantly decreased (from 69.3 +/- 10 to 37.6 +/- 13.8; p < 0.001). Eight major adverse cardiac events (5%) were recorded during the 1-month follow-up, with a significantly lower rate in the VASP-guided group compared with the control group (0% vs. 10%; p = 0.007). There was no difference in the rate of major and minor bleeding (5% vs. 4%; p = 1). CONCLUSIONS: This is the first study to suggest that adjusting the clopidogrel loading dose according to platelet monitoring using the VASP index is safe and may significantly improve the clinical outcome after PCI in patients with clopidogrel resistance despite a first 600-mg loading dose. PMID- 18387445 TI - Will measuring vasodilator-stimulated phosphoprotein phosphorylation help us optimize the loading dose of clopidogrel? PMID- 18387446 TI - Insights into (the interstitium of) degenerative aortic valve disease. PMID- 18387448 TI - Vascular closure devices: begin with the end in mind. PMID- 18387451 TI - Overview of pain management in older persons. AB - The purpose of this article is to situate the social and demographic context of pain management in older adults. It summarizes representative literature on the age and sex composition of pain among older adults and considers sources of assessment bias that likely lead to the conflicting descriptions of prevalence. It also describes treatment options that are both available and acceptable to older adults. PMID- 18387452 TI - Pain perception in the elderly patient. AB - In the elderly patient, the barriers to effective treatment of pain are substantial. Even the perception of pain may differ from that in those of less advanced years. Of course, many other factors impinge on the presence of, and treatment of, pain in elderly patients. Issues of physical accessibility to treatment, cost of drugs, the presence of coexisting illness, the use of concomitant medication, and even the ability to understand the complaints of the patient who has cognitive impairment are only some of those factors that contribute to the complexity of the situation. PMID- 18387453 TI - Assessment of pain in the elderly adult. AB - The goals of a clinical assessment for pain in the elderly adult may be similar to those established for younger patients; however, unique characteristics of aging make this assessment more challenging for clinicians. The overarching goal of pain assessment in the elderly is to provide successful pain management. This article provides the clinician with the foundation to perform a successful pain assessment for older adults who are able to communicate by self-report. This provides a comprehensive base on which to build a relevant plan of care. PMID- 18387454 TI - Assessment of pain in the nonverbal or cognitively impaired older adult. AB - The inability of nonverbal older adults to communicate pain represents a major barrier to pain assessment and treatment. This article focuses on nonverbal older adult populations with dementia, delirium, and severe critical illness. A comprehensive approach to pain assessment is advocated encompassing multiple sources of information. Selected behavioral tools for nonverbal pain assessment are critiqued. Although there are tools with promise, there is currently no standardized behavioral tool that may be recommended for broad adoption in clinical practice and continued concerted effort to this end is needed. PMID- 18387455 TI - Special issues and concerns in the evaluation of older adults who have pain. AB - Older patients who have pain present unique challenges for clinicians. On the one hand, care must be taken to treat the pain aggressively while avoiding hampering the patient with excessive side effects, such as drowsiness, nausea and vomiting, and constipation. On the other hand, the clinician must be aware of the growing problem of prescription drug abuse and assess whether or not the patient or his or her family is at risk. Indeed, the concern for assessment is not solely centered on the patient but also extends to the family and extended support network, which may or may not have the patient's best interests at heart when it comes to pain medications. Supposing that addiction and abuse are solely the purview of the young is no longer acceptable, and we have the burden of assessing for problematic behavior while also trying to convince some patients that they would benefit from pain medicine. PMID- 18387456 TI - Pharmacotherapy of pain in older adults. AB - Pain is a universal part of being human, and yet, there is ample evidence that many people from all backgrounds, stages of life, and levels of health care experience receive less than optimal treatment of their pain. This article reviews the pharmacotherapy of pain in older adults, with a focus on salicylates, nonsteroidal anti-inflammatory drugs, and opioids. PMID- 18387457 TI - Topical analgesic agents. AB - Pain processing and transmission are achieved by a complex interaction of pathways and processes. Those parts of the process with peripheral representation may be amenable to therapeutic intervention by systemic administration to achieve a peripheral effect or by local application, including local topical administration to the skin overlying the painful area. Advantages include high level of patient acceptance, ease of administration, avoidance of systemic side effects, and reduced drug-drug interactions. Those drugs with topical analgesic effects include those with specific topical analgesic indication and others in which no such indication exists but that may offer a chance of pain therapy at reduced risk. PMID- 18387458 TI - Role of rehabilitation medicine in the management of pain in older adults. AB - Pain management may play an important role in contributing to optimal quality of life in the elderly population. Pain lowers overall quality of life in part by decreasing function and by amplifying the psychologicic stress of aging. A comprehensive, multidisciplinary approach to pain management, with preservation and restoration of function in older adults, is the cornerstone of an effective pain management program. PMID- 18387459 TI - Behavioral approaches to pain management in the elderly. AB - Pain is a complex phenomenon, influenced by many individual and external factors, and may be experienced differently with age. The detrimental health and social effects of chronic pain are well known. Age-related disorders, such as dementia, may interfere with the communication of pain. Health care provider bias and cultural expectations also may be barriers to the recognition and management of pain in the elderly. A multidisciplinary and multimodal approach in older adults is essential to effective assessment and management. Behavioral approaches to pain should be considered and incorporated into treatment where appropriate. PMID- 18387460 TI - Interventional techniques for back pain. AB - Most Americans will be afflicted by some form of spine-related pain in their lifetime. In older patients, the most frequent source of back pain is lumbar spinal stenosis or vertebral compression fracture. Although most back pain is self-limited, some patients will require interventional techniques. This article reviews minimally invasive techniques for treating back pain, lumbosacral radicular pain, lumbar spinal stenosis, and compression fractures. PMID- 18387461 TI - Lumbar spinal stenosis in older adults: current understanding and future directions. AB - Lumbar spinal stenosis is a common cause of low back pain in the elderly. Surgical and nonsurgical treatments have been used with varying success. This article aims to characterize the growing unmet need for the treatment of neurogenic intermittent claudication in the elderly population with lumbar spinal stenosis and reviews the current understanding of this condition with an eye toward framing a research agenda for nonsurgical treatments. PMID- 18387465 TI - Sentinel lymph node biopsy is successful and accurate in male breast carcinoma. AB - BACKGROUND: Men and women with breast cancer have similar risks of morbidity related to axillary lymph node dissection (ALND). Sentinel lymph node (SLN) biopsy minimizes this risk. We report results from the largest series of SLN biopsies for male breast cancer and compare this experience with that of female counterparts treated concurrently. STUDY DESIGN: The Memorial Sloan-Kettering Cancer Center SLN biopsy database showed that 7,315 SLN biopsy procedures were performed for primary breast cancer from September 1996 to July 2005. Of these, 78 (1.0%) procedures were performed in men. Followup data were obtained from medical record review. RESULTS: SLN biopsy was successful in 76 of 78 (97%) patients. Negative SLNs were found in 39 of 76 (51%) patients. In 3 (8%) patients with negative SLNs, a positive non-SLN was found, identified by intraoperative palpation. Positive SLNs were found in 37 of 76 (49%) patients. In 22 of 37 (59%), node positivity was determined intraoperatively, prompting immediate ALND. In 15 of 37 (41%) patients with positive SLNs, node positivity was determined postoperatively. Of these 15, 9 (60%) underwent completion ALND. In the 2 of 78 (3%) patients with failed SLN biopsy procedures, ALND was performed and yielded positive nodes. At a median followup of 28 months (range 5 to 96 months), there were no axillary recurrences. Compared with their female counterparts, men with breast cancer had larger tumors and were more likely to have positive nodes. CONCLUSIONS: SLN biopsy is successful and accurate in male breast cancer patients. Although a larger proportion of men have positive nodes, for men with negative nodes, SLN biopsy may reduce morbidity related to ALND. PMID- 18387464 TI - Do variations in provider discussions explain socioeconomic disparities in postmastectomy breast reconstruction? AB - BACKGROUND: The use of postmastectomy reconstruction varies with socioeconomic status, but the etiology of these variations is not understood. We investigated whether these differences reflect variations in the rate or qualitative aspects of the provider's discussion of reconstruction as an option. STUDY DESIGN: Data were collected through chart review and patient survey for stages I to III breast cancer patients during the National Initiative on Cancer Care Quality. Multivariable logistic regression was used to identify predictors of reconstruction and discussion of reconstruction as an option. Predictors of not receiving reconstruction despite a documented discussion were also determined. RESULTS: There were 253 of 626 patients who received reconstruction (40.4%). Younger, more educated Caucasian women who were not overweight or receiving postmastectomy radiation were more likely to receive reconstruction. Patients who were younger, more educated, and not receiving postmastectomy radiation were more likely to have a documented discussion of reconstruction. If a discussion was documented, patients who were older, Hispanic, not born in the US, and received postmastectomy radiation were less likely to receive reconstruction. The greatest predictor of reconstruction was medical record documentation of a discussion about reconstruction. CONCLUSIONS: We observed disparities in the likelihood of reconstruction that were at least partially explained by differences in the likelihood that reconstruction was discussed. But there were also differences in the likelihood of reconstruction based on age, race, and radiation once discussions occurred. Efforts to increase and improve discussions about reconstruction may decrease disparities for this procedure. PMID- 18387466 TI - What constitutes a "high-volume" hospital for pancreatic resection? AB - BACKGROUND: Annual institution resection volume has been proposed for defining centers of excellence, with various cut-offs for defining "high-volume" centers used. This study aimed to define an objective, evidence-based operative volume threshold associated with improved postoperative outcomes after pancreatic resection. STUDY DESIGN: This retrospective analysis of patients who underwent pancreatic resection in the Nationwide Inpatient Sample, a 20% representative sample of patients in the US between 1998 and 2003, was performed using multivariable logistic regression. Different models of annual hospital resection volume were analyzed and the goodness of fit of each "high-volume" model to postoperative mortality was compared through use of the pseudo r(2). RESULTS: Based on analysis of 7,558 patients who underwent pancreatic resection, median annual institution resection volume was 15 (range 1 to 254), and overall in hospital mortality was 7.6%. The best model of "high-volume" centers was an annual institution resection volume of 19 or more, as determined by goodness of fit (r(2) of 5.29%). But there was little difference in data variance explained between this best model and other "high-volume" models. The model without any volume variable had a goodness-of-fit r(2) of 3.57%, suggesting that volume explains less than 2% of data variance in perioperative death after pancreatic resection. CONCLUSIONS: Very little difference was observed in the explanatory powers of models of "high-volume" centers. Although volume has an important impact on mortality, volume cut-off is necessary but insufficient for defining centers of excellence. Volume appears to function as an imperfect surrogate for other variables, which may better define centers of excellence. PMID- 18387467 TI - Predictive factors of deep abdominal complications after operation for hydatid cyst of the liver: 15 years of experience with 672 patients. AB - BACKGROUND: Operations are the mainstay of liver hydatid cyst (LHC) treatment. Operations are still associated with high morbidity and mortality because of specific postoperative complications (bile leaks, bilomas, deep bleeding, and deep suppurations) and deep abdominal complications (DAC). The aim of this study was to identify the predictive factors of DAC after LHC operation. STUDY DESIGN: We conducted a retrospective study of 672 patients with LHC treated at the Surgery Department "A" at Ibn Sina University Hospital, Rabat, Morocco. Specific morbidity (DAC) and 30 variables were assessed. Univariate and multivariate logistic regression were performed to identify predictive factors for DAC. An associated risk scoring system was developed. RESULTS: Six hundred sixty-four patients underwent operations. Mortality rate was 0.8% (n = 5) and DAC rate was 18.4% (n = 121). Five independent predictive factors of DAC after LHC operation were retained, ie, presence of cyst preoperative complications (odds ratio [OR] = 3.10; 95% CI, 1.85 to 5.17), 3 or more cysts in the liver (OR = 2.55; 95% CI, 1.42 to 4.59), thick pericyst (OR = 2.59; 95% CI, 1.27 to 5.29), biliary fistula (OR = 2.27; 95% CI, 1.38 to 3.72), and capitonnage alone as residual cavity management (OR = 2.23; 95% CI, 1.12 to 4.44). Multivariate model showed a good fit. Discriminating ability of the model was fair. In theoretical risk, scores ranged from 0 to 5. When the score was 2 or more, sensitivity of the scoring model was 80.3%, specificity was 58.5%, positive predictive value was 30.3%, and negative predictive value was 93%. CONCLUSIONS: Identification of these five factors will allow more appropriate therapeutic care after LHC operation. PMID- 18387468 TI - Comparison of generic versus specific quality-of-life scales for mesh hernia repairs. AB - BACKGROUND: With the use of mesh shown to considerably reduce recurrence rates for hernia repair and the subsequent improvement in clinical outcomes, focus has now been placed on quality-of-life outcomes in patients undergoing these repairs, specifically, as they relate to the mesh prosthesis. Traditionally, quality of life after hernia surgery, like many other medical conditions, has been tested using the generic SF-36 survey. The SF-36 quality-of-life survey, although well studied and validated, may not be ideal for patients undergoing hernia repairs. We propose a new quality-of-life survey, the Carolinas Comfort Scale (CCS), pertaining specifically to patients undergoing hernia repair with mesh; our goal was to test the validity and reliability of this survey. STUDY DESIGN: The CCS questionnaire was mailed to 1,048 patients to assess its acceptability, responsiveness, and psychometric properties. The survey sample included patients who were at least 6 months out after hernia repair with mesh. Patients were asked to fill out the CCS and the generic SF-36 questionnaires, four questions comparing the two surveys, and their overall satisfaction relating to their hernia repair and mesh. RESULTS: The reliability of the CCS was confirmed by Cronbach's alpha coefficient (0.97). Test-retest validity was supported by the correlation found between two different administrations of the CCS; both Spearman's correlation coefficient and the kappa coefficient were important for each question of the CCS. Assessment of its discriminant validity showed that both the mean and median scores for satisfied patients were considerably lower than those for dissatisfied patients. Concurrent validity was demonstrated by the marked correlations found between the CCS and SF-36 questionnaire scales. When comparing the two surveys, 72% of patients preferred the CCS questionnaire, 80% believed it was easier to understand, 66% thought it was more reflective of their condition, and 69% said they would rather fill it out over the SF-36. CONCLUSIONS: The CCS better assesses quality of life and satisfaction of patients who have undergone surgical hernia repair than the generic SF-36. PMID- 18387469 TI - Postoperative hyperbilirubinemia is an independent predictor of longterm outcomes after cardiopulmonary bypass. AB - BACKGROUND: Two decade-old studies of cardiopulmonary bypass (CPB) patients documented a 25% to 35% incidence of postoperative hyperbilirubinemia, associated with increased in-hospital morbidity and mortality. Longterm consequences of this complication are unknown. STUDY DESIGN: Medical records of CPB patients were reviewed. Mortality was ascertained through the National Death Index. Proportional hazards determined important factors in post-CPB survival. Logistic regression delineated predictors of hyperbilirubinemia. Kaplan-Meier and Mantel Cox log-rank survival analyses compared hyperbilirubinemia groups. RESULTS: Bilirubin levels were followed in 826 (59.7%) patients. Bilirubin was normal in 570 (69.0%) patients (group 1), it was 1.4 to 2.8 mg/dL in 184 (22.3%) patients (group 2), and it exceeded 2.8 mg/dL in 72 (8.7%) patients (group 3). Elevated bilirubin was associated with decreased body mass index, congestive heart failure, heparin before operation, postoperative transfusion requirement, bleeding, and renal failure. In-hospital mortality was 4.3% in group 2 and 25.0% in group 3, compared with 0.9% in group 1 (p<0.001). Two-year crude survival was 95.8% in group 1, 84.8% in group 2, and 62.5% in group 3 (p<0.001). Multivariable predictors of longterm mortality were older age, history of stroke, emergency operation, increased duration of cardiopulmonary bypass, respiratory failure, and elevated bilirubin. Compared with survival in group 1, there was a 1.7-fold decrease in group 2 2-year survival (95% CI 0.9 to 3.0; p=0.09) and a 3.8-fold decrease in group 3 survival (95% CI 2.0 to 7.2; p<0.001). CONCLUSIONS: Postoperative bilirubin elevation in CPB patients is common and deadly. The predictive power of hyperbilirubinemia is similar to that of respiratory failure. The cause of postbypass hyperbilirubinemia is unknown and is probably multifactorial. Additional prospective studies are warranted. PMID- 18387470 TI - Emergency laparoscopic management of perforated sigmoid diverticulitis: a promising alternative to more radical procedures. AB - BACKGROUND: Classic emergency surgical management of complicated perforated sigmoid diverticulitis is based on the principle of a two-stage operation, with colon resection and temporary stoma (Hartmann's procedure). But the later second stage operation can be technically difficult and can be associated with a significant morbidity rate. We argue that laparoscopy may be beneficial in such patients with peritonitis in terms of operative results and could facilitate later surgical management. STUDY DESIGN: We studied all consecutive patients with perforated sigmoid diverticulitis requiring emergency surgery between January 2000 and December 2004. RESULTS: Twenty-four patients underwent emergency laparoscopic management for perforated sigmoid diverticulitis. Nineteen patients (80%) were found to have a purulent or fecal diffuse peritonitis. No conversion and colostomy were necessary. The overall morbidity rate was 8%; 2 patients with pelvic abscesses required radiologic drainage. The median hospital stay was 12 days (range 7 to 35 days). Prophylactic sigmoid resection was performed by laparoscopy in all patients, with a conversion rate of 16%. CONCLUSIONS: Laparoscopic treatment of generalized peritonitis secondary to diverticulitis is feasible and safe and may be a promising alternative to more radical surgery in selected patients, avoiding fecal diversion and allowing a delayed elective laparoscopic sigmoid resection. PMID- 18387471 TI - Age-related differences in diagnosis and morbidity of intestinal malrotation. AB - BACKGROUND: Intestinal malrotation in adulthood may present with a variety of chronic symptoms. Surgical intervention frequently leads to other complications in these patients. We hypothesized that the chronic nature of malrotation in adults could cause a delay in diagnosis and increased perioperative complications. STUDY DESIGN: All patients diagnosed with intestinal malrotation from July 2002 through July 2006 were included. IRB approval was obtained. Outcomes in patients less than 16 years of age were compared with outcomes from those older than 16. Presenting symptoms, initial diagnosis, results of imaging data, and time to diagnosis were evaluated. Surgical management, resulting complications, and rate of reoperation were analyzed. RESULTS: Twenty-four patients with intestinal malrotation were identified (age range, 10 days to 89 years old; 10 adults, 14 children). Seventy percent of adults experienced chronic symptoms for 6 months or more before the diagnosis of malrotation was made (children, 14%, p = 0.017). No patients in the adult group were initially diagnosed with malrotation, although 57% of children were correctly diagnosed at the time of presentation of symptoms (p=0.006). Postoperative complications occurred in 60% of adults, but in only 29% of children, though this did not reach significance (p=0.211). Forty percent of adult patients required reoperation (p=0.020). CONCLUSIONS: Intestinal malrotation in adults is often associated with a delay in diagnosis and increased morbidity. Enhanced awareness of this entity in adults may enhance patient counseling and improve therapeutic outcomes in these patients. PMID- 18387472 TI - Causes of early mortality after laparoscopic adjustable gastric banding. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) is 1 of the most used surgical procedures to treat obesity, with > 120,000 bands positioned around the world. LAGB is considered a safe and technically simple procedure, but it has many longterm complications, such as erosion, bleeding that requires reoperations, and sometimes even death of the patient. STUDY DESIGN: This article reviews the deaths after LAGB reported in the literature. Eight hundred eighty articles were reviewed from 1994 to May 2006, using the PubMed Web site and French, English, and Australian health Web sites. RESULTS: Data from 9,682 patients were collected from 24 articles and 48 (0.51%) deaths were reported. Twenty-eight patients died within 30 days of the operation, including 3 intraoperative deaths; 16 patients died 30 or more days after operation; and in 4 patients, the timing of death was not stated in the articles. Pulmonary embolism was the most frequent cause of death, in 11 (22.9%) patients, followed by myocardial infarction in 6 (12.5%) patients. Nine (18.7%) deaths were directly related to band positioning. Longterm mortality rates from LAGB, or reoperations from inadequate weight loss, could not be assessed. CONCLUSIONS: Although technically simple, LAGB has both short- and longterm complications that can be fatal for patients. We recommend surgeons consider perioperative pulmonary embolism prophylaxis and preoperative coronary evaluations on all patients. PMID- 18387473 TI - Physician reimbursement for general surgical procedures in the last century: 1906 2006. AB - BACKGROUND: The last century has seen revolutionary changes in health-care delivery and treatment of surgical diseases. Equally dramatic has been the changes in health-care economics, including the creation of the Medicare and Medicaid programs in 1965. To better characterize the impact to surgical billing, we have undertaken an analysis of physician fees for common general surgical procedures during the past century. STUDY DESIGN: Five common general surgical procedures were analyzed--inguinal herniorrhaphy, appendectomy, cholecystectomy, thyroidectomy, and repair of abdominal aortic aneurysm. Data for physician fees and reimbursement were obtained from the Illinois Medical Blue Book, the University of Iowa Department of Surgery, and the Centers for Medicare and Medicaid Services. The Consumer Price Index was used to convert all data to 2006 dollars. RESULTS: Trends for billed charges of common surgical procedures during the last century have decreased by an average annual rate of approximately 3.5%. Reimbursement during the past 15 years demonstrates continued decreases for repair of abdominal aortic aneurysm (4.7%), thyroidectomy (3.3%), and cholecystectomy (2.9%), and reimbursement for inguinal herniorrhaphy has been flat and for appendectomy has increased slightly (0.9%). The combined annual decrease in reimbursement for the group of 5 surgical procedures during the past 15 years is 1.4%. By comparison, the average annual change in the price of an automobile during the past century has seen a 1.25% increase. Interestingly, milk prices have had a similar trend as surgical procedures with an average annual decline of 1.5% during the last 15 years, which has been attributed to government support (creating a surplus) and increased productivity. CONCLUSIONS: There has been a substantial decline in payments for general surgical procedures during the past century. The influences of government regulations and increased productivity are likely to continue to apply downward pressure on surgical reimbursement. PMID- 18387474 TI - Workforce projections for hepato-pancreato-biliary surgery. AB - BACKGROUND: We wanted to determine whether current levels of fellowship training in hepato-pancreato-biliary (HPB) surgery will be sufficient to meet demand for HPB procedures in 2020. STUDY DESIGN: The Nationwide Inpatient Sample database for 1988 through 2003 was used to construct projections for the number of HPB procedures that will be performed in the US each year through 2020. Available workforce literature was also used to generate estimates of current and future supply of HPB subspecialists. These demand and supply estimates were then used to construct sensitivity analyses of the mean number of HPB procedures per subspecialist in 2020, depending on the degree of regionalization that exists and the number of fellowship-trained subspecialists needed to train each year to meet projected demand for HPB procedures in 2020. RESULTS: An estimated 16,800 HPB procedures will be performed in 2020, representing a 25% increase during the next 15 years. We estimate that 28 fellowship-trained HPB subspecialists will enter the workforce each year. If half of all HPB procedures are performed by HPB subspecialists in 2020, then the average subspecialist will perform only 14 such procedures that year. If high-volume HPB surgery is defined as 40 procedures per year, and 50% of HPB procedures are performed by high-volume surgeons in 2020, then only 15 fellows need to be trained in HPB surgery each year to meet demand in 2020. CONCLUSIONS: Current levels of fellowship training will result in an excess of HPB subspecialists in 2020. PMID- 18387475 TI - Correlation of multidetector CT findings with splenic arteriography and surgery: prospective study in 392 patients. AB - BACKGROUND: To determine the accuracy of contrast-enhanced multidetector CT (MDCT) in demonstrating splenic vascular injury based on results of splenic angiography and operation. STUDY DESIGN: This institutional review board-approved study included 392 hemodynamically stable blunt trauma patients whose admission MDCTs demonstrated splenic injury. Images were assessed for parenchymal injury grade, hemoperitoneum volume, and evidence of bleeding and nonbleeding splenic vascular injury. Splenic arteriography was performed for high splenic injury grade and splenic vascular injury. Medical records were reviewed to determine arteriographic interpretation, surgery indications and findings, outcomes, and demographics. Sensitivity, specificity, predictive values, and accuracy of MDCT in detecting vascular injury were calculated based on results of arteriography and operation. RESULTS: Splenic vascular injury was seen in 22% of patients (86 of 392) on MDCT. Presence of a vascular injury correlated with the CT-based parenchymal splenic injury grade (p < 0.0001). Active splenic bleeding was associated with subsequent clinical deterioration (p < 0.0001). Overall, MDCT had a sensitivity of 76% (76 of 100); specificity of 90% (95 of 106); negative and positive predictive values of 80% (95 of 119) and 87% (76 of 87), respectively; and accuracy of 83% (171 of 206) in detecting vascular injury compared with reference standards. The success rate of nonoperative management was 96%. CONCLUSIONS: MDCT provides valuable information to direct initial clinical management of patients with blunt splenic trauma by demonstrating both active bleeding and nonbleeding vascular injuries. Not all vascular injuries are detected on MDCT, and splenic angiography is still indicated for high-grade parenchymal injury. PMID- 18387476 TI - Outcomes of immediate vertical rectus abdominis myocutaneous flap reconstruction for irradiated abdominoperineal resection defects. AB - BACKGROUND: Perineal wound complications after chemoradiotherapy and abdominoperineal resection (APR) for anorectal cancer occur in up to 60% of patients, including perineal abscess and wound dehiscence. Vertical rectus abdominis myocutaneous (VRAM) flaps have been used in an attempt to reduce these complications by obliterating the noncollapsible dead space with vascularized tissue and closing the perineal skin defect with nonirradiated flap skin. Many surgeons are reluctant to use VRAM flaps unless primary closure is not possible. STUDY DESIGN: All patients who underwent chemoradiotherapy and APR during a 12 year period at the University of Texas MD Anderson Cancer Center were retrospectively reviewed. Patient, tumor, and treatment characteristics and surgical complications and outcomes were compared between patients who underwent VRAM flap reconstruction of wounds that could have been closed primarily (flap group, n = 35) and those who had primary closure of the perineal wound (control group, n = 76). RESULTS: Overall, there were no significant differences in the incidence of perineal wound complications between the groups; the flap group had a significantly lower incidence of perineal abscess (9% versus 37%, p = 0.002), major perineal wound dehiscence (9% versus 30%, p = 0.014), and drainage procedures required for perineal/pelvic fluid collections (3% versus 25%, p = 0.003) than the control group had. Despite flap harvest and the need for donor site closure in the flap group, there was no significant difference in abdominal wall complications between groups during the study's mean patient followup of 3.8 years. CONCLUSIONS: VRAM flap reconstruction of irradiated APR defects reduces major perineal wound complications without increasing early abdominal wall complications. Strong consideration should be given to immediate VRAM flap reconstruction after chemoradiation and APR. PMID- 18387477 TI - Neoadjuvant chemotherapy in invasive breast cancer results in a lower axillary lymph node count. AB - BACKGROUND: It is essential to have the highest level of confidence in axillary staging assessment. Many surgeons and pathologists believe that fewer lymph nodes are present in axillary dissection specimens of women treated by neoadjuvant chemotherapy. Consequently, the purpose of this study was to compare the lymph node counts of axillary dissection specimens from patients having received neoadjuvant chemotherapy with those of patients treated with primary operation. STUDY DESIGN: A retrospective analysis of a prospective database from our institution identified 283 women with invasive breast cancer who underwent level I and II axillary lymph node dissections. Women from the neoadjuvant chemotherapy group (n=107) were compared with those from the primary surgery group (n=176). The total number of lymph nodes harvested was considered as a continuous variable, but also dichotomized into two categories (< 10 and >or=10). Its correlation with the different variables was analyzed. RESULTS: The median number of lymph nodes retrieved in the neoadjuvant chemotherapy group was 10.0 (range 0 to 38) compared with 12.5 (range 0 to 30) in the control group (p=0.002). There were also significantly more patients with fewer than 10 lymph nodes recovered in the neoadjuvant group (45 versus 28%, p=0.007). Logistic regression showed that neoadjuvant chemotherapy was the only factor associated with retrieval of fewer than 10 lymph nodes. CONCLUSIONS: This study suggests that administration of neoadjuvant chemotherapy to breast cancer patients results in a reduced number of lymph nodes retrieved in the axillary dissection specimens. PMID- 18387478 TI - Current role of therapeutic laparoscopy and thoracoscopy in the management of malignancy: a review of trends from a tertiary care cancer center. PMID- 18387479 TI - Scar treatments: preclinical and clinical studies. PMID- 18387480 TI - Current progress in keloid research and treatment. PMID- 18387481 TI - Patient-care-related telephone communication between general surgery residents and attending surgeons. PMID- 18387482 TI - Correlation between intraoperative colonoscopy and virtual colonoscopy. PMID- 18387483 TI - Crossed fused renal ectopia with calculi. PMID- 18387484 TI - An improved technique for esophagojejunostomy after total gastrectomy with a novel anvil grasping forceps. PMID- 18387485 TI - Laparoscopic right hepatectomy: original technique and results. PMID- 18387486 TI - Near-infrared spectroscopy: potential clinical benefits in surgery. PMID- 18387487 TI - Knee-jerk dogma. PMID- 18387490 TI - Are you the consultant's consultant? PMID- 18387491 TI - Pattern recognition in pediatric and adolescent gynecology--a case for formal education. PMID- 18387492 TI - Comments for "Pattern recognition in pediatric and adolescent gynecology, a case for formal education". PMID- 18387495 TI - Risk of post-endoscopic retrograde cholangiopancreatography pancreatitis and ways to prevent it: old myths, a current need? The case of allopurinol. PMID- 18387496 TI - A new paradigm for increasing use of open-access screening colonoscopy. PMID- 18387497 TI - Persistent heartburn in a patient on proton-pump inhibitor. PMID- 18387499 TI - Obesity is an independent risk factor for hepatocellular carcinoma development in chronic hepatitis C patients. AB - BACKGROUND & AIMS: It is not fully elucidated whether obesity enhances hepatocarcinogenesis in patients with chronic hepatitis C. The aim of this study was to investigate the relationship between body weight and risk of hepatocarcinogenesis in chronic hepatitis C patients. METHODS: We enrolled 1431 patients with chronic hepatitis C who visited our liver clinic between 1994 and 2004, excluding those with hepatocellular carcinoma (HCC) at their visit or with a previous history of HCC. They were divided into 4 groups according to body mass index (BMI): underweight (< or =18.5 kg/m(2), N = 112); normal (18.5 to less than 25 kg/m(2), N = 1023); overweight (25 to less than 30 kg/m(2), N = 265); and obese (>30 kg/m(2), N = 31). We assessed the impact of obesity on the hepatocarcinogenesis adjusted by multivariate Cox proportional hazard regression with other risk factors found significant in univariate analysis. RESULTS: During the follow-up period (mean, 6.1 y), HCC developed in 340 patients, showing cumulative incidence rates of 10.5%, 19.7%, and 36.8% at 3, 5, and 10 years, respectively. The incidence differed significantly among the BMI groups (P = .007). Adjusting for other significant factors, overweight and obesity were shown to be an independent risk factor of HCC, with a hazard ratio of 1.86 (95% confidence interval, 1.09-3.16; P = .022) and 3.10 (95% confidence interval, 1.41 6.81; P = .005) as compared with the underweight patients. CONCLUSIONS: The risk of HCC in patients with chronic hepatitis C increases in proportion to BMI in a wide range of its values, from underweight to obese. PMID- 18387498 TI - Hepatitis C and non-Hodgkin lymphoma among 4784 cases and 6269 controls from the International Lymphoma Epidemiology Consortium. AB - BACKGROUND & AIMS: Increasing evidence points towards a role of hepatitis C virus (HCV) infection in causing malignant lymphomas. We pooled case-control study data to provide robust estimates of the risk of non-Hodgkin's lymphoma (NHL) subtypes after HCV infection. METHODS: The analysis included 7 member studies from the International Lymphoma Epidemiology Consortium (InterLymph) based in Europe, North America, and Australia. Adult cases of NHL (n = 4784) were diagnosed between 1988 and 2004 and controls (n = 6269) were matched by age, sex, and study center. All studies used third-generation enzyme-linked immunosorbent assays to test for antibodies against HCV in serum samples. Participants who were human immunodeficiency virus positive or were organ-transplant recipients were excluded. RESULTS: HCV infection was detected in 172 NHL cases (3.60%) and in 169 (2.70%) controls (odds ratio [OR], 1.78; 95% confidence interval [CI], 1.40 2.25). In subtype-specific analyses, HCV prevalence was associated with marginal zone lymphoma (OR, 2.47; 95% CI, 1.44-4.23), diffuse large B-cell lymphoma (OR, 2.24; 95% CI, 1.68-2.99), and lymphoplasmacytic lymphoma (OR, 2.57; 95% CI, 1.14 5.79). Notably, risk estimates were not increased for follicular lymphoma (OR, 1.02; 95% CI, 0.65-1.60). CONCLUSIONS: These results confirm the association between HCV infection and NHL and specific B-NHL subtypes (diffuse large B-cell lymphoma, marginal zone lymphoma, and lymphoplasmacytic lymphoma). PMID- 18387501 TI - Precut sphincterotomy based on papillary characteristics. PMID- 18387502 TI - Protective effect of Misgurnus anguillicaudatus polysaccharide on immunological liver injury in mice. AB - The modulation influence of Misgurnus anguillicaudatus polysaccharide on the expression of nitric oxide synthase (NOS), B cell lymphoma/leukemia-2 (Bcl-2, hepatocyte apoptosis inhibitor) and Bcl-2 associated X protein (Bax, hepatocyte apoptosis promoter) in mice's liver with immunological hepatic injury was studied. Immunological hepatic injury was induced by lipopolysaccharide (LPS ip, 0.2 mg kg(-1)) in bacillus calmette-guerin (BCG ip, 0.15 g kg(-1), once, before 7 days) primed mice. The mice were treated with M. anguillicaudatus polysaccharides (MAP) at doses of 30 mg kg(-1), 60 mg kg(-1), respectively, ig, once a day, and sacrificed on the 8th day after ip LPS for 4 h. In comparison to the normal mice, the nitric oxide production, serum alanine aminotransferase (sALT) and serum glutathione s-transferase (sGST) levels were increased significantly, iNOS and Bax expression were up-regulated by 16.5 times (P<0.001 vs. normal animal group) and 0.43 times (P<0.05, vs. normal animal group) respectively, cNOS expression was not apparently changed, and no Bcl-2 expression was found in immunological hepatic injury mice. The M. anguillicaudatus polysaccharide (30 mg kg(-1)) could reduce sALT, sGST and nitric oxide production levels (vs. BCG-LPS model control group) by 25.1%, 42.6% and 17.8% respectively, and the expression of iNOS and Bax was decreased (vs. BCG-LPS model control group) by 80.3% and 38.4%, while the expression of cNOS and Bcl-2 increased (vs. BCG-LPS model control group) by 58.7% and 352%, respectively. PMID- 18387503 TI - Regulation of aminopeptidase N (EC 3.4.11.2; APN; CD13) on the HL-60 cell line by TGF-beta(1). AB - Membrane-bound peptidases interfere with cellular growth, differentiation, activation and death by fine-tuning local concentrations of various signaling peptides such as the growth factors, hormones, chemokines and cytokines. We examined the effects of anti-inflammatory cytokine transforming growth factor beta(1) (TGF-beta(1)) on the expression and activity of aminopeptidase N (APN), an ectoenzyme processing several signaling peptides. Myelo-monocytic HL-60 cell line having high basal APN activity corresponding to the membrane CD13 marker served as a model. Regulation of CD13/APN was assayed at the levels of mRNA and at the membrane marker CD13. Functional properties of CD13/APN were examined by measuring the enzyme activity, and the signal transduction ability, followed as Ca(++) mobilization triggered by APN-blocking WM-15 antibody. TGF-beta(1) at physiological concentrations (0.16 to 2.5 ng/mL) increased expression of CD13 both at mRNA and membrane protein level in a time- and concentration-dependent manner. Transcriptional activation of CD13 by TGF-beta(1) is suggested as actinomycin-D, an inhibitor of RNA synthesis, abrogated the TGF-beta(1)-induced up-regulation of CD13. Increased membrane CD13 expression was associated with an increase of its enzyme (APN) activity and with a decrease of its signal transduction ability. Anti-inflammatory cytokine TGF-beta(1) counteracted the effects of pro-inflammatory cytokine IFN-gamma on membrane CD13 expression in a time- and concentration-dependent fashion, suggesting a cytokine-regulated role of CD13/APN in inflammation. This is the first report on regulation of CD13/APN expression by TGF-beta(1) on immature cells of myelo-monocytic origin. As obtained with physiological concentrations of TGF-beta(1) these findings may be relevant for cytokine-regulated CD13/APN expression on mature myeloid cells in the course of inflammation. PMID- 18387504 TI - L.E.A.P.S. heteroconjugate is able to prevent and treat experimental autoimmune myocarditis by altering trafficking of autoaggressive cells to the heart. AB - We evaluated the efficacy of the Ligand Epitope Antigen Presentation System (L.E.A.P.S.trade mark) in preventing or treating experimental autoimmune myocarditis (EAM) in A/J mice. L.E.A.P.S. (here, J-My-1) is a conjugate of the myocarditogenic peptide of cardiac myosin MyHCalpha(334-352) (My-1) and J peptide, derived from the sequence of human beta-2 microglobulin. Remarkably, early prophylactic (J-My-1 injected on days -14 and -7 before EAM induction), late prophylactic (J-My-1 injected on days 0, 7, 14, and 21), and therapeutic (J My-1 injected on days 7, 14, and 21 or 10, 17 and 24) administration of J-My-1 significantly decreased the incidence and severity of EAM. However, extended therapeutic treatment was associated with anaphylaxis and death, corresponding with global immune activation associated with J-My-1 treatment. In J-My1-treated animals, we observed expanded numbers of activated CD69+ and CD44+ CD4+ and CD8+ T cells in the spleens. J-My-1 treatment also increased the proportion of CD11c+ dendritic cells in spleens and induced strong production of anti-J-My-1 specific antibodies. J-My-1 injections resulted in decreased levels of chemokines MIP 1alpha and IP-10 in hearts. We propose that J-My-1 treatment interferes with trafficking of autoaggressive immune cells to the heart. PMID- 18387505 TI - Opioids modulate constitutive B-lymphocyte secretion. AB - The opioid system plays a major role in immunomodulation, while its action on cells of the immune system may be opioid receptor-mediated or not. Opioid effects on B-lymphocytes are considered as indirect, attributed to an interplay between distinct cell populations. The aim of the present study was to investigate whether opioid agonists (morphine, alpha(S1)-casomorphin and ethylketocyclazocine) may have a direct action on the secretion of antibodies and cytokines by multiple myeloma-derived cell lines and normal CD19+ B-lymphocytes. Our results show that opioids modulate antibody and cytokine secretion by multiple myeloma cells in a cell line-dependent and opioid receptor-independent manner, while they decrease antibody secretion by normal B-lymphocytes. Furthermore, they decrease the proliferation rate of multiple myeloma cells through opioid receptor activation. Our data suggest two different mechanisms of action of opioids, mediated by different signaling pathways: an early non-opioid receptor-related effect, modulating the constitutive immunoglobulin and cytokine secretion, and a long-term receptor-mediated action on cell growth. These data suggest a further opioid implication in the control of humoral immunity. PMID- 18387506 TI - Chemical modification of iGb3 increases IFN-gamma production by hepatic NKT cells. AB - Isoglobotrihexosylceramide (iGb3) has been identified as an endogenous ligand recognized by NKT cells; however, it is a weak agonist compared to the exogenous alpha-galactosylceramide. Modification of the structure of iGb3 might improve its stimulatory activity. In this study, we assessed the stimulating activity of chemically-modified iGb3 analogues on murine hepatic NKT cells. We analyzed the percentage of IFN-gamma- or IL-4-producing cells in hepatic iNKT cell population and found that two chemically-modified iGb3 analogues, especially 4'''-dh-iGb3, induced significantly greater intracellular IFN-gamma+ NKT cells in liver by flow cytometry. In vivo experiments also showed that 4-HO-iGb3 and 4'''-dh-iGb3 are selectively strong inducer for rapid serum IFN-gamma production compared with unmodified iGb3. Comparing the structure of iGb3 and its two iGb3 analogues, 4-HO iGb3 has an extra hydroxy group on C4, suggesting that the additional hydroxy group of phytosphingosine might augment the stability of the CD1d/glycoceramide complex forming and thereby possibly promote IFN-gamma producing. By further modifying the polysaccharide of glycolipid as did in 4'''-dh-iGb3, we found that 4'''-dh-iGb3 elicited more Th1-biased responses than iGb3 and 4-HO-iGb3. This modification might more strongly strengthen the affinity of the TCR/glycoceramide complex and ultimately polarize iNKT cells to release more Th1 cytokines. Our data suggests that a combination modification on both polysaccharide and sphingosine chain of iGb3 elicits preferential Th1-biased responses. PMID- 18387507 TI - 17beta-Estradiol enhances expression of inflammatory cytokines and inducible nitric oxide synthase in mouse contact hypersensitivity. AB - The effects of 17beta-estradiol (E2) on the expression of cytokines, inducible nitric oxide synthase (iNOS), and cyclooxygenase (COX)-2 in mouse contact hypersensitivity (CHS) were examined. Three week old female mice were ovariectomized, administered 3.2 microg of E2 subcutaneously, the mice sensitized by application of 4-ethoxymethylene-2-phenyl-2-oxazolin-5-one (OXA) on their backs, and CHS induced by applying OXA to the auricles. E2 significantly increased mRNA expression of interferon (IFN)-gamma and interleukin (IL)-10 in the auricle at 6 and 24 h after allergy elicitation in the ear, respectively, when compared to untreated controls. Although there was no effect of E2 on the expression of IL-4 and COX-2 at any time, the expression of iNOS mRNA was increased by E2 treatment at 48 h after elicitation. E2 also enhanced the expression of tumor necrosis factor (TNF)-alpha and IL-1beta. Histological evaluation revealed that E2 promoted edema of the auricle dermis. The hyperplasia of the epidermis was suppressed by E2 and the cell infiltration observed after elicitation was not altered by E2. These results suggest that E2 enhances the expression of IFN-gamma, TNF-alpha, and IL-1beta to augment the edema of auricle dermis in mouse CHS. PMID- 18387508 TI - Immunomodulatory and radioprotective effects of lignans derived from fresh nutmeg mace (Myristica fragrans) in mammalian splenocytes. AB - Recently, the lignans present in the aqueous extract of fresh nutmeg mace (aril of the fruit of Myristica fragrans) were shown to possess antioxidant properties in cell free systems and protected PUC18 plasmid against radiation-induced DNA damage. The present report describes the immunomodulatory and radiomodifying properties of lignans present in the aqueous extract of fresh nutmeg mace in mammalian splenocytes. These macelignans (ML) inhibited the proliferation of splenocytes in response to polyclonal T cell mitogen concanavalin A (Con A). This inhibition of proliferation was due to cell cycle arrest in G1 phase and augmentation of apoptosis as shown by increase in pre G1 cells. The increase in activation induced cell death by ML was dose dependent. It was found to inhibit the transcription of IL-2 and IL-4 genes in response to Con A. The production of IL-2, IL-4 and IFN-gamma cytokines was significantly inhibited by ML in Con A stimulated lymphocytes in a dose dependent manner. ML protected splenocytes against radiation-induced intracellular ROS production in a dose dependent manner. ML was not cytotoxic towards lymphocytes. On the contrary, it significantly inhibited the radiation-induced DNA damage in splenocytes as indicated by decrease in DNA fragmentation. To our knowledge, this is the first report showing the antioxidant, radioprotective and immunomodulatory effects of lignans in mammalian cells. PMID- 18387509 TI - Hesperidin, hesperidin methyl chalone and phellopterin from Poncirus trifoliata (Rutaceae) differentially regulate the expression of adhesion molecules in tumor necrosis factor-alpha-stimulated human umbilical vein endothelial cells. AB - The fruits of Poncirus trifoliata (L.) are widely used in Oriental medicine to treat allergic inflammation. Recently, several active compounds including hesperidin, hesperidin methyl chalone and phellopterin from P. trifoliata (Rutaceae) were isolated and characterized. The goal of this study was to investigate the differential effect of hesperidin, hesperidin methyl chalone and phellopterin derived from P. trifoliata (Rutaceae) on the induction of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by TNF-alpha and the possible molecular mechanisms by which they differentially regulate ICAM-1 and VCAM-1 expressions. Stimulation of human umbilical vein endothelial cells (HUVECs) with TNF-alpha resulted in the increase of ICAM-1 and VCAM-1 expressions, while pretreatment with the three components completely inhibited VCAM-1 expression in a dose-dependent manner but had no effect on ICAM-1 expression. All three compounds failed to block TNF-alpha induced phosphorylation of ERK1/2, which is involved in regulating ICAM-1 production by TNF-alpha. Furthermore, they efficiently inhibited the phosphorylation of Akt and PKC, suggesting that Akt or PKC pathways are an important target by which these compounds regulate TNF-alpha-induced VCAM-1 but not ICAM-1. Additionally, treatment with these chemicals also inhibited U937 monocyte adhesion to HUVECs stimulated with TNF-alpha. Interestingly, the inhibitory effect of hesperidin, hesperidin methyl chalone and phellopterin on monocyte adhesion to HUVECs was recapitulated by transfecting cells with VCAM-1 siRNA. Taken together, hesperidin, hesperidin methyl chalone and phellopterin reduce TNF-alpha-induced VCAM-1 expression through regulation of the Akt and PKC pathway, which contributes to inhibit the adhesion of monocytes to endothelium. PMID- 18387510 TI - Effects of three anti-TNF-alpha drugs: etanercept, infliximab and pirfenidone on release of TNF-alpha in medium and TNF-alpha associated with the cell in vitro. AB - Tumor necrosis factor-alpha (TNF-alpha) is a vital component of the inflammatory process and its aberrant over-expression has been linked to numerous disease states. New treatment strategies have sought to reduce circulating TNF-alpha, either with neutralizing anti-TNF-alpha binding proteins such as etanercept or via drugs that inhibit de novo TNF-alpha synthesis like pirfenidone. In the present study, we examined the effects of both classes of drugs on secreted and cell-associated TNF-alpha produced by THP-1 cells. All of the tested drugs significantly reduced secreted levels of bioactive TNF-alpha following stimulation with LPS as measured by bioassay. However, etanercept-treated cells had approximately six-fold higher levels of cell-associated TNF-alpha compared with that of the LPS-alone treatment group. Surprisingly, LPS+infliximab treated cells did not increase cell-associated TNF-alpha relative to the LPS-alone treatment. Pirfenidone significantly reduced both secreted and cell-associated TNF-alpha levels. These drug-related differences in cell-associated TNF-alpha may have broad implications in the future for the therapeutic uses of anti-TNF-alpha drugs in the management of TNF-alpha diseases. PMID- 18387511 TI - Immunomodulatory effects of curcumin: in-vivo. AB - Curcumin specifically exhibits cytostatic and cytotoxic effects against tumors of multiple origin. Previously we have demonstrated apoptotic activity of curcumin against tumor cells with no effect on normal cells in-vitro. Many anti-cancer drugs exhibit deleterious effects on immune cells, which restrict their wide use in-vivo. In the present study, we have evaluated the effect of curcumin on the major functions of T cells, natural killer cells, macrophages and on total splenocytes in-vivo, which insight the role of curcumin on their broad effector functions. This study demonstrates that prolonged curcumin-injections (i.p.) do not impair the cytotoxic function of natural killer cells, the generation of reactive oxygen species and nitric oxide from macrophages and the levels of Th1 regulatory cytokines remained unaltered. Interestingly, curcumin-injections enhanced the mitogen and antigen induced proliferation potential of T cells. We have also evaluated immunomodulatory effects of curcumin in ascites-bearing animals. This study strengthens our belief that curcumin is a safe and useful immunomodulator for the immune system. PMID- 18387512 TI - Changes in HLA-DR expression, cytokine production and coagulation following endotoxin infusion in healthy human volunteers. AB - The infusion of a low dose of endotoxin into healthy subjects triggers a complex inflammatory response but the intricacies of which, despite extensive research, are still being unraveled. Nine healthy male volunteers received a dose of 30 Units endotoxin/kg bodyweight as an intravenous bolus. Following endotoxin infusion the concentration of TNF-alpha in their serum rapidly increased within 30 min, peaked after 1-2 h and returned to baseline by 4 h. This corresponded to a similarly rapid increase in anti-inflammatory soluble TNF receptor (sTNFR) levels, which remained elevated for up to 48 h. Increased levels of other cytokines were measured, including IL-6, IL-8, G-CSF, IL-1ra and IL-10. However, these cytokines lagged behind that of TNF-alpha and remained elevated for up to 8 h. Endotoxin injection resulted in complex changes in HLA-DR expression, a marker of monocyte activation state. Initially, following a lag of 2-4 h, HLA-DR expression decreased with a nadir at 8 h, followed by an increase in expression above baseline at 22 h. HLA-DR levels returned to baseline 48 h post-endotoxin challenge. This was in contrast to endotoxin-induced changes in white blood cell (WBC) numbers, which dropped rapidly (at 2-3 h) while HLA-DR levels were stable and then peaked during the nadir in HLA-DR expression (8 h). Furthermore, endotoxin injection caused activation of both fibrinolytic and coagulation pathways. Thus, endotoxin infusion results in complex changes in HLA-DR expression, production of pro- and anti-inflammatory cytokines and activation of coagulation. PMID- 18387513 TI - Modulation of adjuvant arthritis in the rat by 2-methoxyestradiol: an effect independent of an anti-angiogenic action. AB - Angiogenesis is a prominent feature in rheumatoid arthritis. 2-methoxyestradiol (2ME2) inhibits endothelial cell proliferation, and angiogenesis in vivo. We evaluated the effect of 2ME2 in rats with adjuvant arthritis (AA), an autoimmune T-cell-dependent polyarticular arthritis induced by immunization with Mycobacterium organisms. Rats were immunized with Mycobacterium butyricum and arthritis was assessed clinically, by radiolabeled blood neutrophil (PMNL) migration to joints and by histology. Treatment with 2ME2 (30 mg/kg/d or 100 mg/kg/d) from day 6 post-immunization inhibited arthritis severity on day 14 (vehicle clinical score=11.2; 2ME2 groups=7-8, p<0.05). When treatment was delayed until signs of clinical arthritis on day 10 post-immunization, 2ME2 treatment still inhibited arthritis severity. PMNL migration to the joints was significantly inhibited (by 35-40%; p<0.01) by early 2ME2 treatment (day 6-14). Treatment with 2ME2 inhibited PMNL migration to dermal inflammation induced by TNF-alpha but not by LPS or C5a. Joint histology revealed decrease in leukocyte infiltration and especially in cartilage damage. However, synovial vascularity was not affected by 2ME2 treatment. The marked splenomegaly, splenitis and lymphoid hyperplasia associated with AA were prevented by 2ME2 therapy. Furthermore, the ex vivo proliferative response to mycobacterial antigen (PPD) of lymphocytes from 2ME2-treated rats with AA was markedly diminished, although response to mitogens was unaffected. Thus 2ME2 has anti-arthritic properties with a disease-modifying action, separate from its anti-angiogenic properties. The selective inhibition of TNF-alpha-induced leukocyte recruitment, lymphoid hyperplasia and attenuated recall response to antigen suggests both immunomodulatory and anti-inflammatory actions of 2ME2. PMID- 18387514 TI - Immunomodulatory activity of novel adjuvant formulations based on Montanide ISA oil-based adjuvants and peptidoglycan monomer. AB - The aim of the study was to directly compare the potential of Montanide ISA720 and ISA206 oil-based adjuvant formulations on the induction of Th1/Th2-type of immune response, and to compare their effect to Complete Freund's adjuvant (CFA), a well known Th1 inducer. IgG isotype profiles (IgG1/IgG2a ratios) and specific cytokine secretion (IFN-gamma and IL-4) as specific markers of Th1/Th2-type of immune response were monitored in experimentally immunised mice using ovalbumin (OVA) as an antigen. Specifically, we wanted to evaluate whether the incorporation of immunostimulating peptidoglycan monomer (PGM) into two oil-based adjuvants (ISA720(PGM) and ISA206(PGM)) influences their capability on Th1/Th2 type of immune response switching. The experiments were carried out using two genetically different inbred strains of mice, i.e. CBA and NIH/OlaHsd mice, respectively. We found significant differences in immune responses related to the genetic background of the two mice strains used in the study. In both mice strains, ISA720 formulations had similar effect to the positive control, CFA, and induced the switch towards Th1-type of immune response specific for OVA. However, ISA206 formulations were less effective in inducing the switch towards Th1 in CBA mice, while in NIH/OlaHsd mice promoted the switch towards Th2-type of immune response. PMID- 18387515 TI - Continued inhalation of lidocaine suppresses antigen-induced airway hyperreactivity and airway inflammation in ovalbumin-sensitized guinea pigs. AB - It is unclear whether inhaled lidocaine is effective against airway hyperreactivity and inflammation in asthma. The aim of this study was to investigate the effects of inhaled lidocaine on airway hyperreactivity and inflammation. Airway reactivity to inhaled histamine, cellular composition of bronchoalveolar lavage (BAL) fluid, plasma substance P (SP), and isolated lung tissue were evaluated in ovalbumin (OVA)-sensitized guinea pigs 7 days after OVA challenge. The effects of inhaled lidocaine on this model were also evaluated. Treatment with lidocaine was administered in two fashions: as single inhalation or inhalation bid for 7 consecutive days, for comparison with a saline-inhaled control group. Airway hyperreactivity to histamine, increase in number of total cells and increased proportion of eosinophils in BAL fluid, and marked eosinophil infiltration in airway walls were noted even 7 days after OVA challenge in the control group. Plasma SP level was also significantly increased. Although treatment with single lidocaine inhalation did not affect airway hyperreactivity, continued inhalation (bid for 7 days) attenuated airway hyperreactivity. Continued, but not single, inhalation of lidocaine also suppressed infiltration of eosinophils in BAL fluid and in airway walls. In addition, plasma SP levels were significantly reduced by continued but not by single inhalation. It appears possible that lidocaine when inhaled suppresses eosinophilic inflammation of the airway and SP-induced neurogenic inflammation, leading to alleviation of airway hyperreactivity. PMID- 18387516 TI - A comparative study on cannabidiol-induced apoptosis in murine thymocytes and EL 4 thymoma cells. AB - It has been shown that leukemia and glioma cells are sensitive to cannabidiol (CBD)-induced apoptosis, whereas primary monocytes and glia cells are relatively insensitive. In the current study, the cellular events and sensitivity to CBD induced apoptosis between murine thymocytes and EL-4 thymoma cells were compared. Cannabidiol markedly induced apoptosis in a time- and concentration-related manner in both cells. The efficacy of CBD to induce apoptosis was comparable between the 2 types of T cells, whereas CBD induced apoptosis in thymocytes with a slightly greater potency than in EL4 cells. Time-course analyses revealed CBD mediated apoptosis occurred earlier in EL-4 cells than that in thymocytes. An increased level of cellular reactive oxygen species (ROS) was detected in both cells with the peak response at 2 h post CBD treatment. Concordantly, CBD triggered a gradual diminishment in the cellular thiols. The presence of N-acetyl L-cysteine (NAC), a precursor of glutathione, markedly attenuated the induction of apoptosis, and restored the diminished levels of cellular thiols. The results demonstrated that both thymocytes and EL-4 thymoma cells were susceptible to CBD induced apoptosis and that ROS played a critical role in the apoptosis induction. PMID- 18387517 TI - Emodin accelerates osteoblast differentiation through phosphatidylinositol 3 kinase activation and bone morphogenetic protein-2 gene expression. AB - Emodin is a naturally occurring anthraquinone present in the roots and bark of numerous plants of the genus Rhamnus. Here, we identified emodin as one of compounds activating the mRNA expression of bone morphogenetic protein (BMP)-2 in the differentiation process of mouse osteoblastic MC3T3-E1 subclone 4 cells. Without any effect on cell growth, the low concentration (up to 5 microM) of emodin highly induced the mRNA expression of BMP-2, the expression of alkaline phosphatase (an early marker of osteoblast differentiation), and the mineralization. Interestingly, emodin induced the activation of phosphatidylinositol 3-kinase (PI3K), Akt and mitogen-activated protein (MAP) kinases, but those inductions by emodin were completely inhibited by the PI3K inhibitor, LY294002, suggesting that the up-regulation of BMP-2 by emodin could be mediated through the activation of both Akt and MAP kinases by activating PI3K. Additionally, emodin-induced activation of NF-kappaB suggested that NF kappaB might be required for its anabolic activity in part. In conclusion, the use of natural compounds with anabolic activity such as emodin could have a beneficial effect on bone health and this kind of studies further elucidate the pharmacological roles of natural compounds in the prevention of osteoporosis and provide the initiative in the early drug discovery and development for osteoporosis. PMID- 18387518 TI - Inhibition of activated responses in dendritic cells exposed to lipopolysaccharide and lipoteichoic acid by diarylheptanoid oregonin. AB - Antigen-presenting dendritic cells may play an important role in the pathogenesis of inflammatory skin diseases, including atopic dermatitis. Oregonin is demonstrated to have anti-inflammatory and anti-oxidant effects. The present study was designed to assess the effect of oregonin against stimulated responses in dendritic cells of mouse bone marrow and spleen. Dendritic cells exposed to lipopolysaccharide, lipoteichoic acid and IL-1beta exhibited increase in the production of IL-12 p70 and TNF-alpha, increase in the formation of reactive oxygen species and nitric oxide, and elevation of intracellular Ca2+ levels. Treatment of oregonin attenuated the microbial product- or IL-1beta-stimulated responses in dendritic cells in a dose-dependent manner. Oregonin revealed a significant inhibitory effect on the production of cytokine, the formation of reactive oxygen species and nitric oxide, and the change in intracellular Ca2+ levels in dendritic cells of bone marrow and spleen. The results show that oregonin seems to attenuate the stimulated cell responses, including cytokine production, in dendritic cells exposed to microbial products and IL-1beta. The findings suggest that oregonin may exert an inhibitory effect against the dendritic cell-mediated immune response. PMID- 18387519 TI - Establishing the phenotype in novel acute and chronic murine models of allergic asthma. AB - Allergic asthma is a chronic disease of the airways, with superimposed acute inflammatory episodes which correspond to exacerbations of asthma. Two novel models of allergic asthma have been developed in mice receiving the same allergen sensitisation, but with acute or chronic allergen exposures, the latter to mimic the human situation more closely. Ovalbumin-sensitised mice were challenged by ovalbumin inhalation twice on the same day for the acute model, and 18 times over a period of 6 weeks for the chronic model. Lung function was monitored in conscious, unrestrained mice immediately after the last challenge for up to 12 h. Airway responsiveness to inhaled methacholine and serum antibody levels were determined 24 h after challenge. Bronchoalveolar inflammatory cell recruitment was determined at 2 or 24 h. Acute and chronically treated mice had similar early and late asthmatic responses peaking at 2 h and 7-8 h, respectively. IgE and IgG antibody levels, compared with naive mice, and eosinophil infiltration, compared with naive and saline challenge, were elevated. Airway hyperresponsiveness to methacholine was observed 24 h after challenge in both models. The acute model had higher levels of eosinophilia, whereas the chronic model showed hyperresponsiveness to lower doses of methacholine and had higher levels of total IgE and ovalbumin-specific IgG antibodies. Both novel murine models of allergic asthma bear a close resemblance to human asthma, each offering particular advantages for studying the mechanisms underlying asthma and for evaluating existing and novel therapeutic agents. PMID- 18387520 TI - Induction of nitric oxide synthases in primary human cultured mast cells by IgE and proinflammatory cytokines. AB - The mast cells have been suggested to be a cellular source of nitric oxide (NO) which level is increased in the pathogenesis of asthma. However, isoforms of the NO generating enzyme, nitric oxide synthase (NOS), in primary human mast cells have not been defined due to the lack of a suitable model. We hence examined directly the expression of NOS mRNA and proteins in primary human cultured mast cells (HCMC). Mature HCMC were cultured from CD34+ progenitors isolated from buffy coat preparations and were subjected to IgE sensitisation, IgE receptor mediated activation and cytokines induced stimulation. While expression of NOS mRNA was detected by conventional reverse transcription-polymerase chain reaction (RT-PCR) and quantitatively analyzed with real-time RT-PCR, expression of NOS proteins was detected by immunostaining. In non-stimulated HCMC incubated in medium alone, expressions of NOS were not detected. While overnight incubation of HCMC with IgE significantly increased the expression of NOS2 and NOS3, only NOS2 expression was up-regulated after overnight incubation with a mixture of TNF alpha, IFN-gamma and IL-1beta. Cross-linking of IgE with anti-IgE further increased NOS2 expression with a concomitant decrease in NOS3 expression. NOS1 was not detected in all treatments. In conclusion, we have shown for the first time that NOS2 and NOS3 expressions are induced in primary human mast cells following appropriate stimulations. Comparisons between the differential expressions of NOS isoforms in HCMC to the changes in NOS expressions in asthma models suggest that the mast cell is a source of NO in asthmatic airways. PMID- 18387521 TI - Pharmacodynamic monitoring of the conversion from mycophenolate mofetil to enteric-coated mycophenolate sodium in stable kidney-allograft recipients. AB - BACKGROUND: The formulations of mycophenolic acid, i.e., mycophenolate mofetil (MMF) and enteric-coated mycophenolate sodium (EC-MPS), seem to have different pharmacokinetic profiles. The aim of this study was to compare the effects MMF and EC-MPS on T-cell proliferation, T-cell activation, T-cell function, and lymphocyte subsets. CLINICAL STUDY AND METHODS: Ten stable kidney-transplant patients on standard maintenance therapy of tacrolimus and MMF (1 g/d), with or without steroids, were converted from MMF to EC-MPS at equivalent dose (720 mg/d). Tacrolimus and steroid doses remained unchanged before, and at 1, 2, 3, and 6 months (M) after conversion. Intra T-lymphocyte cytokines IL-2 and TNF alpha, lymphocyte-activation surface markers (CD25 and CD71), T-cell proliferation (PCNA+ PI(high)), total lymphocyte count, as well as lymphocytes subsets (CD2, CD3, CD4, CD8, CD19, NK cells) were measured by flow cytometry before conversion and at M1, M2, M3, and M6. RESULTS: We found no significant differences of MMF versus EC-MPS on lymphocyte function. T-cell proliferation and T-cell activation (CD25 and CD71 expression), but not cytokine expression (TNF alpha and IL-2), showed a trend to increase after conversion from MMF to EC-MPS. Total lymphocyte, CD2, CD3, CD4, CD8, and NK cells counts were not significantly modified. CONCLUSION: This study revealed a trend to a lower immunosuppression with EC-MPS as compared to MMF in stable renal transplant patients. PMID- 18387525 TI - Of mice and men - and lopsided birds. AB - The article by Zucca and Sovrano (2008, this issue) represents part of a new wave of studies of lateralization in nonhuman species. This work is often in conflict with earlier studies of human cerebral asymmetry and handedness, and the associated claim that these asymmetries are uniquely human, and perhaps even a result of the "speciation event" that led to modern humans. It is now apparent that there are close parallels between human and nonhuman asymmetries, suggesting that they have ancient roots. I argue that asymmetries must be seen in the context of a bilaterally symmetrical body plan, and that there is a balance to be struck between the adaptive advantages of symmetry and asymmetry. In human evolution, systematic asymmetries were incorporated into activities that probably are unique to our species, but the precursors of these asymmetries are increasingly evident in other species, including frogs, fish, birds, and mammals especially primates. PMID- 18387526 TI - Seasonal anisotropy in handedness. AB - The preference for using either the left or the right hand has been linked to important human characteristics such as language lateralisation within the cerebral hemispheres, and evidence has been reported that the proportions of different types of handedness may be influenced by factors such as levels of maternal hormones and anxiety. Under such influences, it is possible in principle that distributions of handedness provide evidence of seasonal anisotropy, that is, variation in the direction of handedness for births in different parts of the year. The results of a number of studies are compared here, and shown to provide evidence of a significant tendency for the incidence of left-handed people to be higher among those born in the spring and ensuing months (March-July in the northern hemisphere) than among those born in the remainder of the year, at least among the male population. PMID- 18387527 TI - Animal lateralization and social recognition: quails use their left visual hemifield when approaching a companion and their right visual hemifield when approaching a stranger. AB - Quails were tested for leftward and rightward turning preferences in a detour task. When facing a mirror located behind a barrier composed of vertical bars, quails showed a striking population-level preference for turning leftward. In order to check whether the asymmetry reflected a motor or a sensory (i.e. visual hemifield) bias, in a second experiment quails were reared in pairs and then tested in the detour task with a familiar (companion) or an unfamiliar (stranger) conspecific as a target. Quails turned leftward when viewing the stranger, but they turned rightward when viewing the companion. These findings are discussed in relation to current evidence for brain lateralization in response to social stimuli in non-human animals. PMID- 18387528 TI - Differential processing of hierarchical visual stimuli in young and older healthy adults: implications for pathology. AB - Hierarchical figures in which large (global) forms are constructed from smaller (local) forms (Navon, 1977) have proved valuable in studies of perceptual organisation and hemispheric specialisation in both healthy volunteers and a wide range of neurological and psychiatric patients. In studies using Navon figures, normal young adults typically identify global forms faster than local forms. When the global and local forms are incongruent (e.g., a large E made of smaller Rs), global forms often interfere with local form identification more than vice versa. In two conditions on the same subjects, we contrasted the performance of young (mean age 22 years) and older (mean age 58 years) healthy volunteers on global and local processing. In the directed attention task, subjects were instructed to detect a target letter that occurred at the prespecified local or global level. The young subjects showed, as expected, faster reaction times (RTs) to detect global targets. In contrast, the older subjects showed significantly faster RTs to the local targets. Likewise, in a divided attention task, in which subjects were instructed to detect a target letter that could occur at either the local or the global level, the young adults were slightly quicker to detect the global targets and the older subjects were significantly quicker to detect the local targets. Error rates were generally low and there was no significant speed/accuracy trade-off in either condition. The observed local precedence effects in healthy older subjects were unexpected and are discussed in reference to previous work on differential hemispheric aging. That work has suggested that the left hemisphere is preferentially biased toward local processing and ages relatively slowly while the right hemisphere is biased toward global processing and ages relatively quickly. The implications of such putative differential aging for the interpretation of pathological local/global processing in neurological and psychiatric diseases are also emphasised. PMID- 18387529 TI - Functional retrograde amnesia: a multiple case study. AB - Functional retrograde amnesia (RA) is a rare pathology and has been rarely studied in detail across different patients. We extensively examined five functional RA patients and compared their neuropsychological profile including anterograde and retrograde memory performance, executive functions, emotional processing, and formally assessed psychiatric symptoms. Across patients, neuropsychological deficits beyond RA were most consistently seen in executive functions and attention suggesting that these dysfunctions contribute to the remote memory deficit. In a majority of the patients, problems in social cognition and emotional behaviour were reflected in Theory of Mind deficits and accompanying psychiatric symptoms. Aberrances in a measure of social desirability were detected, pointing to repressive tendencies in three out of the five patients. Future studies of functional RA patients may investigate more specifically which frontal-lobe associated (dys-) functions contribute to the memory retrieval deficit. Moreover, studying more closely the interaction between social cognition, repressive personality style and memory inhibition in this disease seems worthwhile pursuing. PMID- 18387530 TI - Visual search for facial expressions of emotion is less affected in simultanagnosia. AB - Evidence in healthy human subjects has suggested that angry faces may be enhanced during spatial processing, perhaps even "popping-out" of a crowd. These contentions have remained controversial, but two recent reports in patients suffering from unilateral spatial neglect have lent some support to these views, suggesting that emotional faces capture attention more efficiently than neutral stimuli in the neglected field. Here, we investigate this phenomenon in a patient suffering from severe Balint's syndrome and consequent simultanagnosia. Using a visual search paradigm, we studied differences in the detection of angry, happy and neutral faces, as well as non-emotional stimuli. Results revealed that emotionally expressive faces, in particular anger, were detected more efficiently than other stimuli. These findings corroborate claims that facial expressions of emotion constitute a specific category of stimuli that attract attention more effectively, and are processed prior to attentional engagement. PMID- 18387531 TI - Cerebellar cognitive affective syndrome without global mental retardation in two relatives with Gillespie syndrome. AB - Although previous studies of Gillespie syndrome have systematically reported a generalized delay of cognitive development (mental retardation or oligophrenia), psychometric data to substantiate this view are strikingly absent. In the present study two first degree relatives (mother and daughter) with Gillespie syndrome were neuropsychologically investigated. Aside from a marked asymmetry in the Wechsler-IQ profile, consisting of significantly better results on the verbal [Verbal IQ (VIQ)] than on the nonverbal part [Performance IQ (PIQ)] of the test, cognitive and behavioral assessments revealed a pattern of abnormalities that closely resembles the "cerebellar cognitive and affective syndrome" (CeCAS) (Schmahmann and Sherman, 1998). Aside from prefrontal dysexecutive dysfunctions such as disturbed cognitive planning and set-shifting, parietal lobe involvement was reflected by impaired visuo-spatial memory and visuo-spatial disorganization in constructional tasks. Within the linguistic domain involvement of the prefrontal and temporal language regions was indicated by impaired letter fluency, incidences of agrammatism, apraxia of speech and disrupted language dynamics. With regard to mood and behavior, a number of personality and affective characteristics were found that are typically associated with prefrontal lobe damage and dysfunction of limbic related regions in the cingulate and parahippocampal gyri. Disinhibited symptoms characterized behavior and affect of the mother while the daughter displayed a variety of inhibited symptoms. As a result, behavioral and cognitive findings in these patients do not support the prevailing view of a global mental retardation as a cardinal feature of Gillespie syndrome but primarily reflect cerebellar induced neurobehavioral dysfunctions following disruption of the cerebrocerebellar anatomical circuitry. PMID- 18387532 TI - Impaired search for orientation but not color in hemi-spatial neglect. AB - Patients with hemi-spatial neglect have trouble finding targets defined by a conjunction of visual features. The problem is widely believed to stem from a high-level deficit in attentional deployment, which in turn has led to disagreement over whether the detection of basic features is also disrupted. If one assumes that the detection of salient visual features can be based on the output of spared 'preattentive' processes (Treisman and Gelade, 1980), then feature detection should remain intact. However, if one assumes that all forms of detection require at least a modicum of focused attention (Duncan and Humphreys, 1992), then all forms of search will be disrupted to some degree. Here we measured the detection of feature targets that were defined by either a unique color or orientation. Comparable detection rates were observed in non-neglected space, which indicated that both forms of search placed similar demands on attention. For either of the above accounts to be true, the two targets should therefore be detected with equal efficiency in the neglected field. We found that while the detection rate for color was normal in four of our five patients, all showed an increased reaction time and/or error rate for orientation. This result points to a selective deficit in orientation discrimination, and implies that neglect disrupts specific feature representations. That is, the effects of neglect on visual search are not only attentional but also perceptual. PMID- 18387533 TI - Season of birth and parental age in right, mixed and left handers. AB - Bulgarian secondary school students (264 right handers, 246 mixed handers and 360 left handers) were investigated concerning subject's month of birth and concerning his mother's and father's age at his birth. Neither the month of birth nor any group of months turned out to be significantly associated with handedness. Similarly, no association between parents' age and handedness was found. PMID- 18387534 TI - The ability to assess muscular force in asymmetrical Parkinson's disease. AB - We tested the ability of eight Parkinson's disease (PD) patients with clearly asymmetrical right-sided motor signs and eight control subjects to assess different levels of muscular forces. In Experiment 1, subjects had first to produce a target-force with one hand (the reference hand) with the assistance of visual feedback, and then match that force with the other hand (the matching hand) without any visual feedback. In Experiment 2, they had to produce a target force with one hand and then estimate it by attributing a numerical value. In Experiment 1, the results showed that PD patients could normally reach the target forces with the more affected left hand but they were impaired in inter-manual force transfer. They were also impaired, in Experiment 2, in estimating forces produced by their more affected hand. Our findings suggest that PD patients present a deficit in sensing motor effort. Effort awareness might be mediated by the basal ganglia. PMID- 18387535 TI - Wheel-chair driving improvement following visuo-manual prism adaptation. AB - Prism adaptation has been used for several years to improve several aspects of unilateral neglect. Parameters ranging from the classical neuropsychological tests to mental imagery or to tactile extinction have been successfully ameliorated following a brief period of adaptation to wedge prisms shifting the visual field to the right. However the potential therapeutic implications of this technique depend on the investigation of more functional and ecological parameters. Here we describe a patient with left hemiplegia and unilateral neglect who was impaired during wheel-chair navigation in the clinical unit. Following a brief adaptation period, this patient showed a sudden improvement of wheel-chair driving as well as of classical tests. The potential implications of prism adaptation for the rehabilitation of unilateral neglect are highlighted by the long duration of improvement obtained after a single adaptation session. PMID- 18387536 TI - Handedness assessment in studies of seasonal anisotropy. PMID- 18387537 TI - Are results from different techniques mutually exclusive in the study of how the brain processes visual search? PMID- 18387538 TI - Tartuferi's retina. PMID- 18387539 TI - The neural correlates of visual mental imagery: an ongoing debate. AB - The functional mechanisms and the neural correlates of visual mental imagery (the faculty whereby we can use our "mind's eye" to visualize objects in their absence) are at the centre of a lively debate in cognitive neuroscience. Neurocognitive models have proposed a functional equivalence between visual perception and visual mental imagery, which would be subserved by common neural substrates, such as the retinotopic areas in the occipital lobe. However, brain damaged patients may demonstrate either impaired imagery and preserved perception, such as the classical Charcot and Bernard case and the patients described by Moro et al. (2008, this issue), or the opposite pattern of performance, consisting of preserved imagery and impaired perception. This double dissociation provides a strong challenge to models postulating a functional and anatomical equivalence of perception and imagery, and suggests that these functions have partly distinct neural correlates. PMID- 18387540 TI - Selective deficit of mental visual imagery with intact primary visual cortex and visual perception. AB - There is a vigorous debate as to whether visual perception and imagery share the same neuronal networks, whether the primary visual cortex is necessarily involved in visual imagery, and whether visual imagery functions are lateralized in the brain. Two patients with brain damage from closed head injury were submitted to tests of mental imagery in the visual, tactile, auditory, gustatory, olfactory and motor domains, as well as to an extensive testing of cognitive functions. A computerized mapping procedure was used to localize the site and to assess the extent of the lesions. One patient showed pure visual mental imagery deficits in the absence of imagery deficits in other sensory domains as well as in the motor domain, while the other patient showed both visual and tactile imagery deficits. Perceptual, language, and memory deficits were conspicuously absent. Computerized analysis of the lesions showed a massive involvement of the left temporal lobe in both patients and a bilateral parietal lesion in one patient. In both patients the calcarine cortex with the primary visual area was bilaterally intact. Our study indicates that: (i) visual imagery deficits can occur independently from deficits of visual perception; (ii) visual imagery deficits can occur when the primary visual cortex is intact and (iii) the left temporal lobe plays an important role in visual mental imagery. PMID- 18387541 TI - Strategic aspects of young, healthy older adults', and Alzheimer patients' arithmetic performance. AB - Forty young adults, 40 healthy older adults, and 23 probable AD patients were asked to solve simple subtraction problems (e.g., 9-3; 14-9) in a choice condition and in a no-choice condition. Participants could choose between retrieval and non-retrieval strategies on each problem in the choice condition and were required to use retrieval on all problems in the no-choice condition. Results showed that arithmetic performance and strategy use were influenced by problem, participant, and strategy characteristics. Age-related differences were found in strategy use and strategy execution. Dementia-related differences were found in strategy execution, but not in strategy selection. AD patients had poorer performance (i.e., larger response times and percent of errors) than age related controls, with especially low accuracy under no-choice condition. The findings have implications for our understanding of aging effects in arithmetic, strategic variations in Alzheimer's patients, and sources of cognitive decline during early stages of Alzheimer's disease (AD). PMID- 18387542 TI - Generation of co-speech gestures based on spatial imagery from the right hemisphere: evidence from split-brain patients. AB - It has been claimed that the linguistically dominant (left) hemisphere is obligatorily involved in production of spontaneous speech-accompanying gestures (Kimura, 1973a, 1973b; Lavergne and Kimura, 1987). We examined this claim for the gestures that are based on spatial imagery: iconic gestures with observer viewpoint (McNeill, 1992) and abstract deictic gestures (McNeill, et al. 1993). We observed gesture production in three patients with complete section of the corpus callosum in commissurotomy or callosotomy (two with left-hemisphere language, and one with bilaterally represented language) and nine healthy control participants. All three patients produced spatial-imagery gestures with the left hand as well as with the right-hand. However, unlike healthy controls and the split-brain patient with bilaterally represented language, the two patients with left-hemispheric language dominance coordinated speech and spatial-imagery gestures more poorly in the left-hand than in the right-hand. It is concluded that the linguistically non-dominant (right) hemisphere alone can generate co speech gestures based on spatial imagery, just as the left-hemisphere can. PMID- 18387543 TI - Generalised script sequencing deficits following frontal lobe lesions. AB - Neuropsychological investigations have consistently shown that frontal cortices are relevant in processing temporal and sequential features of actions. However, one of the main theoretical issues that has been discussed in the last 25 years is whether these brain areas store some abstract representations of actions or, conversely, act upon action representations stored within other posterior associative cortices. We administered to 19 patients with frontal lobe lesions and 19 normal controls, script sequencing and generating tasks concerning actions, natural events and "epoques" (ordered events such as the days of the week). The main findings from frontal lobe patients were a generalised sequencing deficit concerning actions and natural phenomena (and not what we labelled "epoques") with almost intact ability to verbally generate, from the long-term memory, scripts' sequences. These findings are discussed within two of the main theoretical frameworks on frontal lobes: the "processing" perspective of the Supervisory Attentional System, SAS (Norman and Shallice, 1986) and the "representational" one represented by the Structure Event Complex (SEC) theory (Grafman, 2002). PMID- 18387544 TI - Recollection of public events in healthy people: a latent-variable stochastic approach to disentangling retrieval and storage. AB - Recollection of media-mediated past events was examined in 96 healthy participants to investigate the interaction between the age of the subject and the "age" of memories. The results provided evidence that people older than 75 years recall recent events significantly worse than remote ones. Younger participants (47-60 years old) showed the reverse pattern. The implementation of a Markov chains latent-variable stochastic model suggested that reduced efficiency of retrieval rather than storage processes accounts for these results. The findings were interpreted with reference to models of memory trace consolidation, assuming that memory for past public events is dependent on hippocampal structures. PMID- 18387545 TI - Cut the coda: early fluency intervals predict diagnoses. AB - The aim of this study was threefold: (i) to clarify whether letter and category fluency tap different cognitive abilities; (ii) to make diagnostic comparisons and predictions using temporally resolved fluency data; (iii) to challenge and test the widely made assumption that 1-min sum scores are the fluency test measure of choice in the diagnosis of dementia. Scores from six 10-sec intervals of letter and category fluency tests were obtained from 240 participants including cognitive levels ranging from mild subjective cognitive complaints to Alzheimer's disease. Factor analysis revealed clearly separate factors corresponding to letter and category fluency. Category fluency was markedly impaired in Alzheimer's disease but not in Mild Cognitive Impairment. Only scores from relatively early intervals predicted Alzheimer's disease and Mild Cognitive Impairment. The conclusions are (i) letter and category fluency are different tests, category fluency being the best diagnostic predictor; (ii) it would be possible to administer category fluency tests only for 30 sec, because after this point the necessary differential diagnostic information about the patient's word fluency capacity has already been gathered. PMID- 18387546 TI - Retrograde memory after unilateral stroke. AB - Over the past two decades, there has been a growing interest in understanding the neural underpinnings of memory of the past. Numerous patients with retrograde amnesia after acute brain damage have been described, but often the causative lesions are bilateral and/or fairly diffuse and one question that has arisen is whether a unilateral lesion is sufficient to cause retrograde memory impairment. In addition, the impact of lesion side and site on the material specificity and temporal extent of retrograde memory deficits has remained unclear. We set out to investigate these issues by comparing 20 patients who had recently had a unilateral stroke that involved (but was not necessarily limited to) either the frontal or temporal lobe to a group of 10 matched normal control subjects on tests of memory of events and semantic details from the autobiographical and public domains. Results indicated that a unilateral lesion was sufficient to cause significant retrograde memory impairment, with right-sided lesions affecting recall of autobiographical events more than left-sided lesions. The memory deficits in these patients were most often relatively mild, but temporally pervasive rather than characterised by a traditional temporal gradient. Furthermore, memory of events (both autobiographical and public) was impaired in patients who had had a stroke that included the hippocampus, but not in those whose strokes spared this region. Finding that patients with mesial temporal lesions had difficulty remembering details related to public events, even when offered recognition choices, raises the possibility that part of their memory storage network (and not just their retrieval abilities) was compromised. PMID- 18387547 TI - Cerebral blood flow regulation during cognitive tasks: effects of healthy aging. AB - Aging is associated with frontal subcortical microangiopathy and executive cognitive dysfunction, suggesting that elderly individuals may have impaired metabolic activation of cerebral blood flow to the frontal lobes. We used transcranial Doppler (TCD) ultrasound to examine the cerebral blood flow response to executive control and visual tasks in the anterior and posterior cerebral circulations and to determine the effects of healthy aging on cerebral blood flow regulation during cognitive tasks. Continuous simultaneous anterior cerebral artery (ACA) and posterior cerebral artery (PCA) blood flow velocities (BFVs) and mean arterial pressure (MAP) were measured in response to word stem completion (WSC) and a visual search (VS) task in 29 healthy subjects (14 young, 30+/-1.5 years; 15 old, 74+/-1.4 years). We found that: (1) ACA and PCA blood flow velocities are both significantly increased during WSC and VS cognitive tasks, (2) ACA and PCA activations were task specific in our young volunteers, with ACA>PCA BFV during the WSC task and PCA>ACA BFV during the VS task, (3) while healthy elderly subjects also had PCA>ACA BFV during the VS task, they did not have ACA>PCA activation during the WSC task, and (4) healthy elderly subjects tend to have overall greater increases in BFV during both cognitive tasks. We conclude that TCD can be used to monitor cerebrovascular hemodynamics during the performance of cognitive tasks. Our data suggest that there is differential blood flow increase in the ACA and PCA in young versus elderly subjects during cognitive tasks. PMID- 18387548 TI - Variability in cognitive presentation of Alzheimer's disease. AB - The aim of the present study was to explore the nature and prevalence of phenotypic variations in Alzheimer's disease (AD). Neuropsychological profiles of a large cross-sectional cohort of patients with a clinical diagnosis of the disease were examined. All tests distinguished the AD group from controls confirming their sensitivity to the presence of early AD. Factor analysis of test scores revealed five factors, reflecting the discrete cognitive domains of memory, language, perceptuospatial abilities, executive skills, and praxis. Cluster analysis revealed distinct performance profiles that could not be accounted for by disease severity. Some patients showed an accentuation of memory impairment relative to other domains, whereas others showed relative sparing. Cognitive deficits other than memory were the salient presenting feature in a relatively high proportion of patients. A subset of the cohort (22%) showed grossly disproportionate impairments in one cognitive domain. The findings emphasise variability in presentation and indicate that distinct phenotypic variations appear to lie on a continuum rather than representing discrete forms of disease. PMID- 18387549 TI - Spatial attentional bias in normal people: object or viewer-centered. AB - When bisecting radial lines, normal subjects often have a distal bias. To help in explaining this systematic bias, it has been proposed that normal people have an attentional bias to the top of the lines (object centered hypothesis) or toward stimuli falling in the lower portion of the retina (retinotopic hypotheses) or to distal peripersonal space (body centered hypotheses). The primary aim of this study is to test these hypotheses by having normal subjects bisect radial lines, placed in a clockwise distribution in the transverse plane, below eye level. Our results demonstrate that the perception of the midpoint progressively changes as a function of the body centered orientation of the lines, with subjects demonstrating a progressively greater distal bias as they approached the midsagittal plane or radial condition. These findings provide support for the postulate that distal bias on radial lines bisections is body centered. PMID- 18387550 TI - Line bisection as an early method to assess homonymous hemianopia. AB - Horizontal line bisection task is a common clinical task well known to most neuropsychologists. Typically, patients with visuospatial neglect show a reliable ipsilesional deviation in the bisection of long lines. Less well known in the English literature is the typical line bisection error observed in hemianopic patients who show the opposite deviation. In fact, this contralesional deviation in bisection was well known in the old German scientific literature. In 1894, more than 110 years ago, the German physician Dr. D. Axenfeld published a short case report about line bisection as a "simple method to diagnose hemianopia". His paper is one (if not the first) historical report, describing the "typical hemianopic line bisection error". At the time of its publication, it was a very popular paper in the German scientific community frequently cited by subsequent researchers. Between 1900 and 1920, Axenfeld's observation motivated several further studies using bisection by well-known researchers such as Best, Liepmann, Wilbrand, Poppelreuter and Fuchs. Surprisingly, most of today's clinical and cognitive studies use experimental modifications of line bisection in neglect patients and healthy subjects, often without realizing that this task was originally devised for the assessment of hemianopic patients. Consequently, the hemianopic line bisection error was "neglected" for many decades until its recent "rediscovery". The present paper has three aims. First, Axenfeld's classical report is translated. Second, interpretations arising from early bisection studies (around 1900-1930) in hemianopic patients are summarized and framed within contemporary science. Finally, we attempt to explain why this formerly well-known clinical phenomenon was forgotten later for nearly a century. PMID- 18387551 TI - Cajal's retina. PMID- 18387553 TI - A reappraisal of person recognition and identification. AB - In the last years an increasing number of cases (Gainotti et al., 2008, this issue) have been reported in whom difficulty to recognise and identify familiar people occurs in everyday multimodal settings, differently from unimodal face specific impairments (i.e., prosopagnosia). A reappraisal of current person processing models is presented in order to account for such deficits as well as for the common slips of recognition occurring in healthy subjects. The model we propose is based upon three main modifications of current models, namely: (1) the role of PINs as stores of multimodal perceptual knowledge; (2) the richness of perceptual nuances characterizing PINs of most familiar people; (3) the PINs' addressing of Exemplar Semantics by a provisional Gestalt guessing and an analytical check, to be negotiated whenever a conflict arises. A single case report of Capgras delusion is presented as a crossmodal person processing disorder in everyday settings for whom the proposed model allows a cognitive interpretation. PMID- 18387554 TI - Cross-modal recognition disorders for persons and other unique entities in a patient with right fronto-temporal degeneration. AB - We describe a patient (CD), with a right fronto-temporal degeneration, who showed massive defects in the recognition of familiar people and severe behavioural disorders. CD scored in the normal range on tests of episodic memory, attention and visual-spatial abilities, and obtained mildly abnormal scores on naming and executive functions tests. CD was then studied and matched with a left brain damaged patient (IG), comparable to her as for education, naming score and general cognitive impairment, on several tasks, exploring recognition of familiar people and of other instances of 'unique entities'. On specific tasks of face recognition, she obtained normal results on perceptual tests, but highly pathological scores on mnesic-associative tasks. A similar defect was found when identification was based on the person's voice or on a verbal definition. The cross-modal nature of CD's disorder was confirmed by results of a test, in which person-specific information available from photographs and from names was directly compared. In order to evaluate if CD's recognition disorder: (a) was the consequence of a general semantic defect, (b) was specific for people, or (c) also concerned other instances of 'unique entities', we matched her capacity to name and recognize the pictures of items belonging to various categories of knowledge with those concerning famous monuments and famous people. CD identified items belonging to semantic categories much better than those considered as 'unique entities' and, within the latter, obtained slightly better results with famous monuments than with famous persons. MRI showed a bilateral atrophy of the antero-inferior parts of the temporal lobes, more pronounced in the right side. About 2 years after the onset of the symptomatology, CD became untestable, due to the development of a severe motor neuron disease. PMID- 18387555 TI - Categorical and coordinate spatial representations within object-location memory. AB - An important aspect of spatial memory is the ability to remember the positions of objects around us. There is evidence that spatial information can be represented in different ways, involving a coordinate representation (fine-grained, metric information) and a categorical representation (above/below, right/left relations). The current study is aimed at investigating possible lateralization effects for categorical and coordinate information when encoding position information alone and when integrating position information and object information in memory. Twenty-five patients who had suffered from a stroke and 36 healthy controls were tested with different tests assessing categorical and coordinate position memory, and categorical and coordinate object-to-position memory. The identity task that was used by (Laeng, 1994) was included as well as a control task for measuring lateralization effect for categorical and coordinate information. Moreover, object-recognition and visuo-spatial perception were assessed. The results showed that processing categorical and coordinate spatial information were impaired by a lesion in the left and right hemisphere, respectively. No lateralization effects were found when spatial information had to be integrated with object information. These results bear on the functional components of object-location memory and their underlying hemispheric basis. PMID- 18387556 TI - Task-dependent and task-independent neurovascular responses to syntactic processing. AB - The neural basis for syntactic processing was studied using event-related fMRI to determine the locations of BOLD signal increases in the contrast of syntactically complex sentences with center-embedded, object-extracted relative clauses and syntactically simple sentences with right-branching, subject-extracted relative clauses in a group of 15 participants in three tasks. In a sentence verification task, participants saw a target sentence in one of these two syntactic forms, followed by a probe in a simple active form, and determined whether the probe expressed a proposition in the target. In a plausibility judgment task, participants determined whether a sentence in one of these two syntactic forms was plausible or implausible. Finally, in a non-word detection task, participants determined whether a sentence in one of these two syntactic forms contained only real words or a non-word. BOLD signal associated with the syntactic contrast increased in the left posterior inferior frontal gyrus in non-word detection and in a widespread set of areas in the other two tasks. We conclude that the BOLD activity in the left posterior inferior frontal gyrus reflects syntactic processing independent of concurrent cognitive operations and the more widespread areas of activation reflect the use of strategies and the use of the products of syntactic processing to accomplish tasks. PMID- 18387557 TI - The representation of homophones: more evidence from the remediation of anomia. AB - This paper compares two theoretical positions regarding the mental representation of homophones: first, that homophones have one phonological word form but two grammatical representations (lemmas, e.g., Levelt et al., 1999; Dell, 1990), or second, that they have two separate phonological word forms (e.g., Caramazza et al., 2001). The adequacy of these two theoretical accounts for explaining the pattern of generalisation obtained in the treatment of homophone naming in aphasia is investigated. Two single cases are presented, where phonological treatment techniques are used to improve word retrieval. Treatment comprised picture naming of one member of a homophone pair using a phonological cueing hierarchy. A significant improvement in word retrieval was found for both the treated and the untreated homophones, while there was no improvement for phonologically and semantically related controls. It is argued that the data support a shared representation for homophones at the word form level. However, current theories cannot explain the pattern of generalisation found without the addition of a mechanism for repetition priming (e.g., suggested by Wheeldon and Monsell, 1992) and feedback between word form and lemmas to explain the results. PMID- 18387558 TI - Script representation in patients with Alzheimer's disease. AB - We examined script representation in 26 patients with Alzheimer's disease (AD) compared to 31 healthy elderly subjects (HE). Participants were asked to sort cards describing actions belonging to eight scripts according to the script to which they belonged and according to their order of execution. Each script included actions which were low in centrality and distinctiveness (non-central actions and non-distinctive actions--NCA & NDA), and which were high in centrality (central actions--CA), distinctiveness (distinctive actions--DA), centrality and distinctiveness (central actions and distinctive action--CA & DA). These actions were presented in three conditions. In the first condition (scripts with headers--SH), the 43 actions belonging to three different scripts were given with each script header written on separate cards. The second condition (scripts without headers--SwH) used 46 actions belonging to three other scripts, but no script header was provided. In the third condition (scripts with distractor header--SDH), the 28 actions belonging to two other scripts were given with each script header and a distractor header written on separate cards. The results showed that performance of subjects with AD was significantly lower in all conditions. Overall, AD patients made significantly more sequencing errors than HE subjects. AD patients also committed significantly more sorting errors than HE subjects for all types of actions (NCA & NDA, CA, DA, CA & DA). These data are consistent with the view that AD produces impairment of both the syntactic and semantic dimensions of script representation. PMID- 18387559 TI - Intrusions in story recall: when over-learned information interferes with episodic memory recall. Evidence from Alzheimer's disease. AB - Patients with Alzheimer's disease (AD) suffer from distortions of memory. Among such distortions, intrusions in memory tests are frequently observed. In this study we describe the performance of a group of mild AD patients and a group of normal controls on the recall of three different types of stories: a previously unknown story, a well-known fairy-tale (Cinderella), and a modified well-known fairy-tale (Little Red Riding Hood is not eaten by the wolf). The aim of our study was to test the hypothesis that in patients who tend to produce intrusions, over-learned information interferes with episodic recall, i.e., the retrieval of specific, unique past episodes. AD patients produced significantly more intrusions in the recall of the modified fairy-tale compared to the recall of the two other stories. Intrusions in the recall of the modified fairy-tale always consisted of elements of the original version of the story. We suggest that in AD patients intrusions may be traced back to the interference of strongly represented, over-learned information in episodic memory recall. PMID- 18387560 TI - Phonological recoding, visual short-term store and the effect of unattended speech: evidence from a case of slowly progressive anarthria. AB - Phonological processing and short-term memory were investigated in a patient with slowly progressive anarthria. The patient, who had an auditory-verbal span in the lower unimpaired range, showed preserved phonological similarity and word length effects with auditory presentation. These phonological effects of immediate retention were absent with visual input. The patient was also unable to perform phonological judgments on written verbal material. Under unattended speech interference her visual span improved, with an increase of visual confusion errors. In the light of a model including a main auditory-verbal storage component (the phonological short-term input store, STS), and a rehearsal process, that, after phonological recoding, conveys visually presented verbal material to the phonological STS, the patient's pattern of impairment is interpreted as a selective deficit in the process of phonological recoding. This prevents visual-verbal material from accessing the phonological STS, that is, therefore, insulated from visual inputs. PMID- 18387561 TI - A case of selective impairment of encyclopaedic numerical knowledge or 'when December 25th is no longer Christmas day, but '20+5' is still 25'. AB - This study investigates encyclopaedic numerical knowledge in a patient with a presumed left temporal dysfunction, associated with temporal lobe epilepsy. Encyclopaedic numbers are those used as nominal labels (such as in 'British Broadcasting Corporation - BBC 1' or 'Levis 501') to express familiar or historical dates (e.g., our birthday or the French revolution, 1789) and to indicate other general or autobiographical numerical information (e.g., Personal Identification numbers - PINs, post-codes, telephone numbers). We showed a dissociation between impaired processing of encyclopaedic numbers and preserved processing of non-encyclopaedic numbers (e.g., the larger between 54 and 65 or the result of '6 x 9'). This dissociation complements the existing data showing the reverse pattern of performance, namely an advantage for encyclopaedic compared to non-encyclopaedic numbers. These data add important information on an aspect of numerical processing that has not yet been systematically explored and reinforce the distinction between numerical and non-numerical knowledge in the semantic system. PMID- 18387562 TI - The role of the prefrontal cortex in sentence comprehension: an rTMS study. AB - Using repetitive transcranial magnetic stimulation (rTMS), we investigated the role of the left and right dorsolateral prefrontal cortex (DLPFC) in sentence comprehension. Subjects were required to judge which of the two pictures correctly matched the meaning of active and passive semantically reversible sentences (subject-verb-object); the incorrect picture did not match the sentence in term of lexical items (semantic task) or agent-patient structure (syntactic task). The subjects performed the task while a series of magnetic stimuli were applied to the left or right DLPFC. When rTMS was applied to the left DLPFC, the subjects' performance was delayed only for the semantic task, while rTMS applied to the right DLPFC slowed the processing of syntactic information. The results of this experiment provide direct evidence of a double dissociation between the rTMS effects and the type of task, which may reflect a differential hemispheric involvement of working memory resources during sentence comprehension. PMID- 18387563 TI - Automatic memory processes in normal ageing and Alzheimer's disease. AB - This study examined the contribution of automatic and controlled uses of memory to stem completion in young, middle-aged and older adults, and compared these data with a study involving patients with Alzheimer's disease (AD) who performed the same task (Hudson and Robertson, 2007). In an inclusion task participants aimed to complete three-letter word stems with a previously studied word, in an exclusion task the aim was to avoid using studied words to complete stems. Performances under inclusion and exclusion conditions were contrasted to obtain estimates of controlled and automatic memory processes using process-dissociation calculations (Jacoby, 1991). An age-related decline, evident from middle age was observed for the estimate of controlled processing, whereas the estimate of automatic processing remained invariant across the age groups. This pattern stands in contrast to what is observed in AD, where both controlled and automatic processes have been shown to be impaired. Therefore, the impairment in memory processing on stem completion that is found in AD is qualitatively different from that observed in normal ageing. PMID- 18387564 TI - Incomplete ipsilesional hallucinations in a patient with neglect. PMID- 18387565 TI - Numbers, space, and action - from finger counting to the mental number line and beyond. PMID- 18387566 TI - Mind the gap between both hands: evidence for internal finger-based number representations in children's mental calculation. AB - At a certain stage of development, virtually all children use some kind of external finger-based number representation. However, only little is known about how internal traces of this early external representation may still influence calculation even when finger calculation ceases to be an efficient tool in mental calculation. In the present study, we provide evidence for a disproportionate number of split-five errors (i.e., errors with a difference of +/-5 from the correct result) in mental addition and subtraction (e.g., 18 - 7 = 6). We will argue that such errors may have different origins. For complex problems and initially also for simple problems they are due to failure to keep track of 'full hands' in counting or calculation procedures. However, for simple addition problems split-five errors may later also be caused by mistakes in directly retrieving the result from declarative memory. In general, the present results are interpreted in terms of a transient use of mental finger patterns - in particular the whole hand pattern - in children's mental calculation. PMID- 18387567 TI - Does finger training increase young children's numerical performance? AB - Butterworth (1999) suggested that fingers are important in representing numerosities. Furthermore, scores on a finger gnosis test are a better predictor of numerical performance up to 3 years later than intellectual measures (Marinthe et al., 2001; Noel, 2005). We hypothesised that training in finger differentiation would increase finger gnosis and might also improve numerical performance. Accordingly, 47 first-grade children were selected and divided into 3 groups: children with poor finger gnosis who followed the finger differentiation training programme (G1), a control-intervention who were trained in story comprehension (G2), and a group with high finger gnosis scores who just continued with normal school lessons (G3). The finger training consisted of 2 weekly sessions of half an hour each, for 8 weeks. Before the training period, children in G3 performed better in finger gnosis and enumeration than children in the two other groups. After the training period this pattern remained for the children in G2 and G3, but the children in G1 were significantly better than those in G2 at finger gnosis, representation of numerosities with fingers, and quantification tasks; they also tended to be better at the processing of Arabic digits. These results indicate that improving finger gnosis in young children is possible and that it can provide a useful support to learning mathematics. Such an approach could be particularly appropriate for children with a developmental Gerstmann syndrome. Theoretically, these results are important because they suggest a functional link between finger gnosis and number skills. PMID- 18387569 TI - Finger counting habits modulate spatial-numerical associations. AB - This study explored the contribution of finger counting habits to the association of numbers with space (the SNARC effect). First, a questionnaire study indicated that two-thirds of 445 adults started counting on their left hand, regardless of their handedness. Secondly, a group of 53 "left-starters" but not a group of 47 "right-starters" showed a SNARC effect in a parity task. A significant difference in the strength of the effect between groups suggests that finger counting habits indeed contribute to the association between numbers and space in adults. PMID- 18387568 TI - A developmental fMRI study of nonsymbolic numerical and spatial processing. AB - This functional magnetic resonance imaging (fMRI) study systematically investigates whether there is a neurofunctional overlap of nonsymbolic numerical and spatial cognition in (intra)parietal regions in children and adults. The study also explores the association between finger use and (nonsymbolic) number processing across development. Twenty-four healthy individuals (12 children, 12 adults) were asked to make nonsymbolic numerical and spatial (experimental tasks) as well as color discriminations (control task). Using identical stimulus material across the three tasks disentangled nonsymbolic number representations from general attentional mechanisms, visual-spatial processing and response selection requirements. In both age groups, behavioral distance effects were obtained upon processing numerical (but not spatial and/or color) stimuli. Baseline imaging effects revealed age-dependent, partly overlapping activations of nonsymbolic numerical and spatial processing in the right posterior superior parietal lobe (PSPL) in adults only. Interestingly, regions more activated in children relative to adults were centred on bilateral supramarginal gyrus (SMG) and lateral portions of the anterior intraparietal sulcus (IPS), further extending to adjacent right post- and precentral gyrus, the latter of which has been reported to support grasping previously (Simon et al., 2002). Overall, our results are first evidence for an age-dependent neurofunctional link between areas supporting finger use and nonsymbolic number processing and furthermore, might be suggestive of a special role of fingers for the development of number magnitude representations and early arithmetic. PMID- 18387570 TI - On the relationship between handedness and hand-digit mapping in finger counting. AB - The purpose of the current investigation was to evaluate hand-digit mapping in finger counting in French children and adults and whether handedness might constraint hand-digit mapping. To this aim, hand-digit mapping used when counting from one to ten by means of fingers, together with performance-based and preference-based measures of handedness, were evaluated in French individuals of four different age groups (4-5 years old, 6-7 years old, 10-11 years old and 24 47 years old). Irrespective of the age group, analyses revealed a strong tendency to use first the right hand to count from one to five and then the left hand to count from six to ten. In addition, a significant interaction between hand-digit mapping and hand preference was found, with participants who used first their right hand to count reporting higher right-hand preference in unimanual activities. These findings are discussed in light of recent studies assuming a link between finger-counting habits and numerical processing. PMID- 18387571 TI - Digits affect actions: the SNARC effect and response selection. AB - The SNARC (spatial-numerical association of response codes) effect refers to the finding that processing digits can modulate response times, with low digits facilitating left responses and high digits facilitating right responses. Recent evidence indicates that the locus of this effect is in the response selection stage. To examine this possibility, we presented participants with low or high digits and then asked them to make one of two keypresses (left or right) - whichever one they felt more comfortable making. The results showed that low digits biased the selection of left keypresses while high digits biased the selection of right keypresses. Thus, the SNARC effect not only affects how fast responses can be initiated but also affects what responses will be selected. PMID- 18387572 TI - Sensory and cognitive processes of shifts of spatial attention induced by numbers: an ERP study. AB - The relationship between space and number has become a focus of intensive investigation (Hubbard et al., 2005; Walsh, 2003). The present paper aims to explore the nature of attentional shifts induced by the perception of irrelevant numbers as it was shown by Fischer et al. (2003). We measured the event related potentials induced by the perception of visual lateralized targets cued by numbers that differed in their magnitude. Congruent trials were defined as those where a target presented in the Right Visual Field (RVF) followed a large number and those where a target presented in the Left Visual Field (LVF) followed a small number. Numbers generate a modulation of evoked potentials on targets as soon as 80 msec after the presentation of the target: congruency of the target determined the amplitude on perceptual P100 and cognitive P300 in both sides of presentation of the target. Although a typical distribution of the components was found, effects of congruency were distributed around anterior and Centro-Parietal sites. Due to the functional properties of the mentioned components, the present data suggest that, in fact, perception of numbers does affect the location of attention to external space. Moreover, the distribution of the congruency effect signals so that the representational nature of numbers makes a difference with respect to the stimuli classically used in cueing studies of visual attention to location. The role of top-down control generated by numbers is discussed. PMID- 18387573 TI - Time course of number magnitude interference during grasping. AB - In the present study, we recorded the kinematics of grasping movements in order to measure the possible interference caused by digits printed on the visible face of the objects to grasp. The aim of this approach was to test the hypothesis that digit magnitude processing shares common mechanisms with object size estimate during grasping. In the first stages of reaching, grip aperture was found to be larger consequent to the presentation of digits with a high value rather than a low one. The effect of digit magnitude on grip aperture was more pronounced for large objects. As the hand got closer to the object, the influence of digit magnitude decreased and grip aperture progressively reflected the actual size of the object. We concluded that number magnitude may interact with grip aperture while programming the grasping movements. PMID- 18387574 TI - Spatial representations of numbers in children and their connection with calculation abilities. AB - Evidence for a connection between number and space processing comes from behavioural, patient, and brain imaging data, but only a few studies have addressed this issue in children. We asked children (n=118) at the age of 8-9 years to decide which one of the two numerical distances in a visually presented number triplet was numerically larger. Numerical and spatial distances were manipulated independently, resulting in congruent, neutral, and incongruent conditions. The spatial distances between the numbers clearly affected the comparison of numerical distances: reactions times were faster and error rates smaller for congruent than for incongruent trials. These findings are in line with the assumption of a spatial layout of mental number representations in third graders. Correlations between the size of the congruity effect and calculation abilities were found to be differently marked for girls and boys: a positive correlation was found for boys, while a marginally negative correlation was obtained for girls. PMID- 18387575 TI - Impairments of the mental number line for two-digit numbers in neglect. AB - Humans represent numbers along a left-to-right oriented Mental Number Line (MNL). Neglect patients seem to neglect the left part of the MNL, namely the smaller numbers within a given numerical interval. However, until now all studies examining numerical representation have focussed on single-digit numbers or two digit numbers smaller than 50. In this study, the full range of two-digit numbers was assessed in neglect patients and two control groups. Participants were presented with number triplets (e.g., 10_13_18) and asked whether or not the central number is also the arithmetical middle of the interval. The factors manipulated were decade crossing (e.g., 22_25_28 vs 25_28_31), distance to the arithmetical middle (e.g., 18_19_32 vs 18_24_32), and, most importantly, whether the central number was smaller or larger than the arithmetical middle (e.g., 11_12_19 vs 11_18_19). Neglect patients differed from controls in that they benefited less when the middle number was smaller than the arithmetical middle of the interval. Neglect patients thus seem to have particular problems when accessing the left side of numerical intervals, also when adjusted to two-digit numbers. Such an impaired magnitude representation in neglect seems to have detrimental effects on two-digit number processing as the helpful spatial metric of magnitude cannot be properly activated. PMID- 18387576 TI - Basic numerical processing in left intraparietal sulcus (IPS) acalculia. AB - We report a case of a patient, AD, who suffered from acalculia following an infarct restricted to the left IPS (intraparietal sulcus). AD showed deficits in an arithmetic battery and in basic numerical processing tasks. Compared to matched controls, AD showed a larger distance effect in numerical comparisons, an abnormal size congruity indicated by a lack of facilitation by irrelevant numerical values in physical comparisons, a deficiency in counting in comparison tasks, and a deficiency in subitizing in number naming tasks and comparison tasks. This suggests that the underlying deficit in AD is a difficulty in the perception and manipulation of quantity. Moreover, it indicates the essential role of the IPS in basic numerical processing. PMID- 18387577 TI - Lost in number space after right brain damage: a neural signature of representational neglect. AB - The human brain represents numbers along a mental number line, whose spatial nature was confirmed by studies of patients with visuospatial neglect. Here we describe a neural signature of neglect for the left "number space" by using a task that does not require manipulation of numbers. Patients were asked to discriminate an infrequent ("one" or "nine") from a frequent spoken number word ("five"). P3b brain waves, elicited by infrequent stimuli and indexing cognitive processing, were delayed to targets on the left of the number line ("one") compared to targets on the right ("nine"). The delay of P3b is thus a neural signature of the disorder of representational space. PMID- 18387578 TI - Horizontal spatial representations of time: evidence for the STEARC effect. AB - It is well known that stimuli such as numerals (small vs large) and auditory pitches (low vs high) have spatial characteristics, and that responses to such stimuli are biased by the mental representation of their magnitude. Walsh (2003) has argued that any spatially and action-coded magnitude will yield a relationship between magnitude and space. Here we investigated the spatial representation of 'time' using speeded responses to the onset timing (early vs late) of a probe stimulus following periodic auditory clicks. Participants pressed one of the two response keys depending on whether the timing of a given probe was earlier or later than expected based on the preceding clicks. The results showed that left-side responses to early onset timing were faster than those to late onset timing, whereas right-side responses to late onsets were faster than those to early onsets when the response keys were aligned horizontally. Such a time-response congruity effect was not observed with the vertical alignment of responses. These results suggest that time is represented from left to right along the horizontal axis in space. The existence of a 'mental time line' in space and the spatial-temporal association of response codes (STEARC) effect are discussed. PMID- 18387580 TI - Mental representation: what can pitch tell us about the distance effect? AB - Reaction time (RT) profiles for comparing magnitudes (e.g., numbers, physical sizes) are similar - the larger the difference between the compared stimuli, the shorter the RT (distance effect). Nevertheless, it is unclear whether such correspondence is due to similar, two-dimensional, linear mental representations of magnitudes. In contrast, pitch perception has a more complex, two-dimensional, helical representation. This study examined whether comparisons of music pitches are similar to other magnitude response functions. Experiment 1 employed a comparison task, resulting in an RT profile identical to that obtained when comparing other magnitudes. In contrast, Experiment 2 employed a discrimination task, resulting in RTs that matched the helical representation and were dissociated from the classical distance effect. Experiment 3 replicated the results of Experiment 1 using a comparison task with different stimuli and intervals. These findings imply that the distance effect under comparison tasks might reflect a general sensorimotor transformation, rather than mental representation per se. PMID- 18387579 TI - Dissociation of numerosity and duration processing in the left intraparietal sulcus: a transcranial magnetic stimulation study. AB - A possible dissociation of duration and numerosity processing was tested in an off-line repetitive transcranial magnetic stimulation (rTMS) design. Participants had to compare the numerosity of flashed dot sequences or the duration of single dot displays before and after 15 min of 1 Hz rTMS over one of three sites (the left or right intraparietal sulcus (IPS), or the vertex chosen as a control site). Compared to the control site, performance was only slowed down for the numerosity comparison task after the left IPS stimulation, whereas it was not affected for the duration comparison task for any of the parietal sites. These results show that the parietal area critically involved in numerosity processing is not involved in duration processing, revealing at least one cerebral site where duration and numerosity comparison processes dissociate. PMID- 18387581 TI - Selection, preparation, and monitoring: current approaches to studying the neural control of action. PMID- 18387582 TI - Acquisition of a new bimanual coordination pattern modulates the cerebral activations elicited by an intrinsic pattern: an fMRI study. AB - Intensive practice of a new complex motor skill results in progressive improvement of performance. This induces neuroplastic changes, reflecting the transition from attention-demanding to more automatic performance throughout the learning. In the present fMRI study, learning-related cerebral activation changes during the acquisition of a new complex bimanual coordination pattern were examined, i.e., the 90 degrees out-of-phase pattern (90Phi). Furthermore, we investigated whether practice of this new pattern influenced the neural correlates associated with performance of a preferred intrinsic pattern. Twelve young healthy subjects were intensively trained on the 90Phi task, and underwent two fMRI scanning sessions in early (PRE) and late (POST) learning. Scanning sessions included performance of the trained 90Phi pattern, as well as the nontrained intrinsic in-phase pattern (InPhi). Kinematics registered during training and scanning experiments showed that the new 90Phi pattern was acquired successfully, resulting in learning-related brain activation changes. Activation decreases were observed in the right prefrontal cortex (DLPFC and dorsal premotor), in the right middle temporal and occipital cortices and in the posterior cerebellum. Conversely, increases were found in the basal ganglia and hippocampus. Interestingly, activity elicited by the InPhi task also evidenced within-subjects PRE/POST differences (although kinematics InPhi performance was equivalent in both sessions). In particular, the learning-related decreases found for the 90Phi pattern in the cerebellum, the occipital and temporal gyri were similarly observed for the intrinsic InPhi pattern. Moreover, InPhi performance induced PRE/POST increases of activity in the left superior frontal gyrus. Our fMRI results suggest that intensive practice of a new complex coordination pattern impacted, at least temporarily, on the neural correlates of preferred intrinsic coordination patterns. Additional neural recruitment might reflect increased mental effort to prevent negative transfer from the learned mode onto the intrinsic coordination mode. PMID- 18387583 TI - Motor imagery: a window into the mechanisms and alterations of the motor system. AB - Motor imagery is a widely used paradigm for the study of cognitive aspects of action control, both in the healthy and the pathological brain. In this paper we review how motor imagery research has advanced our knowledge of behavioral and neural aspects of action control, both in healthy subjects and clinical populations. Furthermore, we will illustrate how motor imagery can provide new insights in a poorly understood psychopathological condition: conversion paralysis (CP). We measured behavioral and cerebral responses with functional magnetic resonance imaging (fMRI) in seven CP patients with a lateralized paresis of the arm as they imagined moving the affected or the unaffected hand. Imagined actions were either implicitly induced by the task requirements, or explicitly instructed through verbal instructions. We previously showed that implicitly induced motor imagery of the affected limb leads to larger ventromedial prefrontal responses compared to motor imagery of the unaffected limb. We interpreted this effect in terms of greater self-monitoring of actions during motor imagery of the affected limb. Here, we report new data in support of this interpretation: inducing self-monitoring of actions of both the affected and the unaffected limb (by means of explicitly cued motor imagery) abolishes the activation difference between the affected and the unaffected hand in the ventromedial prefrontal cortex. Our results show that although implicit and explicit motor imagery both entail motor simulations, they differ in terms of the amount of action monitoring they induce. The increased self-monitoring evoked by explicit motor imagery can have profound cerebral consequences in a psychopathological condition. PMID- 18387584 TI - Delay-related cerebral activity and motor preparation. AB - Flexible goal-oriented behavior requires the ability to carry information across temporal delays. This ability is associated with sustained neural firing. In cognitive terms, this ability has often been associated with the maintenance of sensory material online, as during short-term memory tasks, or with the retention of a motor code, as during movement preparation tasks. The general issue addressed in this paper is whether short-term storage of sensory information and preparation of motor responses rely on different anatomical substrates. We used functional magnetic resonance imaging (fMRI) to measure sustained and time varying delay-related cerebral activity evoked during performance of a delay non match to sample (DNMS) task, where task contingencies rather than explicit instructions ensured that either sensory or motor representations were used to cross the delay period on each trial. This approach allowed us to distinguish sensory from motor characteristics of delay-related activity evoked by task contingencies, rather than differences in the control of short-term storage driven by verbal instructions. Holding sensory material online evoked both sustained and time-varying delay-related activity in prefrontal regions, whereas movement preparation evoked delay-related responses in precentral areas. Intraparietal cortex was sensitive to the presence of memoranda, but indifferent to the type of information that was retained in memory. Our findings indicate that short-term storage of sensory information and preparation of motor responses rely on partially segregated cerebral circuits. In the frontal lobe, these circuits are organized along a rostro-caudal dimension, corresponding to the sensory or motor nature of the stored material. PMID- 18387585 TI - Dissociations between motor-related EEG measures in a cued movement sequence task. AB - Different aspects of preparation, especially processes related to knowing what to prepare versus applying that foreknowledge effectively, may be reflected in different types of brain activity, e.g., the lateralized readiness potential (LRP), beta-band event-related desynchronization and phase locking. In a previous study in which subjects had to switch between response hands, dissociations were found between types of preparatory hand-related lateralization in evoked potentials, amplitude and phase locking (Gladwin et al., 2006) knowing what task set to switch to and effectively preparing that task set affected the different measures of lateralization. Similarly to how, in task switching, stimuli and responses must be correctly related to each other, in the preparation of movement sequences relations must be specified concerning motor processes. Similar dissociations as found in the task switching data might then be found in a cued movement sequence task. This possibility was explored by precueing elements of a two-movement sequence involving the left and right index fingers, and comparing various measures of electroencephalogram activity. Cues could specify the full sequence, either the first or the second element, or neither element. Knowing the first element was sufficient to lateralize the pattern of phase locking, but effects were found in the LRP and lateralized amplitude only when the full sequence was known. It seems likely that subjects only fully prepared the first response when they had full knowledge of the sequence so that the dissociation may be closely related to that found for task switching. Thus, the present data would appear to agree with previous results that couple response-foreknowledge with phase locking and the transformation of that foreknowledge into effective changes in component processes with evoked potential shifts. The results further underscore the general importance of considering different types of brain activity: depolarization, desynchronization and phase locking all appear to be involved in different aspects of cognitive control. PMID- 18387586 TI - Switching between colors and shapes on the basis of positive and negative feedback: an fMRI and EEG study on feedback-based learning. AB - A crucial element of testing hypotheses about rules for behavior is the use of performance feedback. In this study, we used fMRI and EEG to test the role of medial prefrontal cortex (PFC) and dorsolateral (DL) PFC in hypothesis testing using a modified intradimensional/extradimensional rule shift task. Eighteen adults were asked to infer rules about color or shape on the basis of positive and negative feedback in sets of two trials. Half of the trials involved color-to color or shape-to-shape trials (intradimensional switches; ID) and the other half involved color-to-shape or shape-to-color trials (extradimensional switches; ED). Participants performed the task in separate fMRI and EEG sessions. ED trials were associated with reduced accuracy relative to ID trials. In addition, accuracy was reduced and response latencies increased following negative relative to positive feedback. Negative feedback resulted in increased activation in medial PFC and DLPFC, but more so for ED than ID shifts. Reduced accuracy following negative feedback correlated with increased activation in DLPFC, and increased response latencies following negative feedback correlated with increased activation in medial PFC. Additionally, around 250msec following negative performance feedback participants showed a feedback-related negative scalp potential, but this potential did not differ between ID and ED shifts. These results indicate that both medial PFC and DLPFC signal the need for performance adjustment, and both regions are sensitive to the increased demands of set shifting in hypothesis testing. PMID- 18387587 TI - Dorsal anterior cingulate cortex integrates reinforcement history to guide voluntary behavior. AB - Two competing types of theory have been proposed about the function of dorsal anterior cingulate cortex (dACC): evaluative theories hold that dACC monitors ongoing behavior to detect errors or conflict, whereas response selection theories hold that dACC is directly involved in the decision making process. In particular, one response selection theory proposes that dACC utilizes reward prediction error signals carried by the midbrain dopamine system to decide which of several competing motor control systems should be given control over the motor system (Holroyd and Coles, 2002). The theory further proposes that the impact of these dopamine signals on dACC determines the amplitude of a component of the event-related brain potential called the error-related negativity (ERN). In the present study, we applied this theory to a decision making problem that requires participants to select between two response options in which an erroneous choice is not clearly defined. Rather, the reward received for a particular response evolves in relation to the individual's previous behavior. We adapted a computational model associated with the theory to simulate human performance and the ERN in the task, and tested the predictions of the model against empirical ERP data. Our results indicate that ERN amplitude reflects the subjective value attributed by each participant to their response options as derived from their recent reward history. This finding is consistent with the position that dACC integrates the recent history of reinforcements to guide voluntary choice behavior, as opposed to evaluating behaviors per se. PMID- 18387588 TI - A hand and a field effect in on-line motor control in unilateral optic ataxia. AB - Patients with bilateral optic ataxia fail to show rapid perturbation-induced corrections during manual aiming movements. Based on this, it has been proposed that this pathology results from a disruption of processes of on-line motor control in the posterior parietal cortex (PPC). Here, we show that on-line motor control performance in a patient with unilateral optic ataxia is similar to that of pointing towards stationary targets in peripheral vision, showing the same combination of hand and field effects. We also show that in the patient, manual correction towards his ataxic field was possible only when a preceding saccade (100msec earlier) rapidly provides foveal information about the new target location. In control subjects, manual correction was often, but not necessarily preceded by a saccade. These results allow us to put forward a model of visuo manual transformation, which involves updating of the reach plan based on the target-eye error, and rely upon two dissociated spatial representations (of the hand and of the target, respectively) within the PPC. PMID- 18387589 TI - Action monitoring in major depressive disorder with psychomotor retardation. AB - Major depressive disorder (MDD) is characterized by disturbances of mood and affect, but also by a distinct pattern of psychomotor and cognitive deficits such as motor retardation and impaired executive functioning. An important aspect of executive functioning is performance monitoring, i.e., a continuous checking whether intended action goals have been reached and whether correction of the applied strategy is necessary. A well-known marker for action monitoring is the error negativity (Ne) or error-related negativity (ERN), an event-related potential (ERP) component generated in the anterior cingulate cortex (ACC) following erroneous responses. To date, Ne/ERN amplitudes have been investigated in moderately depressed patients only. The present study is the first to investigate action monitoring in severely depressed patients (mean Hamilton score=28.4). In addition, the patients' psychomotor performance was assessed to see whether there is a relationship between action monitoring and psychomotor retardation. Behavioural and ERP measurements were obtained during performance on a speeded two-choice reaction task in 26 patients with MDD and 25 healthy, matched controls. Psychomotor performance measures were speed of simple movements in various psychomotor tasks and the score on the Salpetriere retardation rating scale (SRRS). Relative to the controls, the patients' behavioural results revealed a similar, but slower performance pattern. Overall between-group differences were demonstrated for the error positivity (Pe) amplitudes, but not for the Ne/ERN amplitudes. However, correlations of the Ne/ERN amplitude with several psychomotor variables were strong. In the depressed patients taking benzodiazepines an additional attenuation of Ne/ERN amplitudes was observed. Only severely depressed patients manifesting retardation showed impeded action monitoring. The correlations between action monitoring and psychomotor performance indicate that in MDD these two processes are highly interdependent, both being deregulated. Moreover, the same network of brain regions is likely to be implicated in both processes. PMID- 18387590 TI - Fast responders have blinders on: ERP correlates of response inhibition in competition. AB - Recent studies have demonstrated that individuals acting in a social context form shared representations, resulting in incorporating another person's action plan into their own. The present study investigated the extent to which shared representations are formed in a competitive task. Specifically, it was tested whether in competition the process of response inhibition is affected by explicit knowledge of another's task. Event-related potential (ERP) correlates of response inhibition were measured while pairs of participants competed with each other on a speeded go/no-go task. Participants were instructed to always try to respond faster than their direct competitor. No-go stimuli requiring an inhibitory response of the other person as well (compatible action) or no-go stimuli to which the other person should respond (incompatible action) were directly compared. Behavioral performance measures and response inhibition, as reflected in the no-go P3, were decreased on incompatible actions compared to compatible ones. Interestingly, both the behavioral and the ERP effects were caused by the slow responding and thus unsuccessful competitors. These findings indicate that shared representations are formed in competitive tasks, but differently for successful and unsuccessful competitors. Only the slow responders are impeded by incompatible actions. The present study therefore demonstrates that the formation of shared representations is not a fully automatic process. People can differ in the extent to which they incorporate the other's action plan into their own and this may be closely related to successful performance in competitive action. PMID- 18387591 TI - Behavioral and cortical mechanisms for spatial coding and action planning. AB - There is considerable evidence that the encoding of intended actions in visual space is represented in dynamic, gaze-centered maps, such that each eye movement requires an internal updating of these representations. Here, we review results from our own experiments on human subjects that test the additional geometric constraints to the dynamic updating of these spatial maps during whole-body motion. Subsequently, we summarize evidence and present new analyses of how these spatial signals may be integrated with motor effector signals in order to generate the appropriate commands for action. Finally, we discuss neuroimaging experiments suggesting that the posterior parietal cortex and the dorsal premotor cortex play selective roles in this process. PMID- 18387592 TI - Imaging causal interactions during sensorimotor processing. AB - We review three ways in which transcranial magnetic stimulation (TMS) has been used to investigate causal interactions between different brain areas: (1) combined with functional imaging; (2) combined with EEG; and (3) applied successively over two brain areas. We discuss the theoretical advantages of each technique, illustrated by examples from the literature, and highlight the potential of these approaches to provide novel insights into the neural bases of action control. PMID- 18387593 TI - Muscle imaging: mapping responses to transcranial magnetic stimulation with high density surface electromyography. AB - Representations of different body parts or muscles in the human primary motor cortex overlap extensively. At the effector level, most muscles are surrounded by and overlap with several neighbours as well. This hampers the assessment of excitability in individual muscles with transcranial magnetic stimulation (TMS), even if so-called "focal" stimulating coils are used. Here we used a novel mapping paradigm based on high-density surface electromyography (HD-sEMG) to investigate the spatial selectivity of TMS in the forearm musculature. In addition, we tested the hypothesis that selective stimulation can be improved by a voluntary background contraction of the target muscle. We mapped and compared the topographies of motor evoked potential (MEP) amplitudes during rest and during background contractions of two forearm muscles (extensor carpi radialis and extensor digitorum communis). The MEP topographies were also compared to the amplitude topography of voluntary EMG. The results indicate that under many conditions a large proportion of the MEP activity recorded at the surface originated from the target muscle's neighbours. There was a systematic relationship between TMS intensity and the topographic distribution of MEP responses during voluntary contraction. With increasing stimulus intensity, the MEP topography deviated increasingly more from the topography of voluntary EMG. We conclude that when standard EMG montages are used, the recorded MEPs are not necessarily evoked in the target muscle alone. Stimulation during a voluntary background contraction of the target muscle may enhance the selectivity of TMS. It however remains essential to use stimulus intensities as low as possible, to minimize the contribution of surrounding non-target muscles to the MEP. PMID- 18387594 TI - CC2D2A, encoding a coiled-coil and C2 domain protein, causes autosomal-recessive mental retardation with retinitis pigmentosa. AB - Autosomal-recessive inheritance is believed to be relatively common in mental retardation (MR), although only four genes for nonsyndromic autosomal-recessive mental retardation (ARMR) have been reported. In this study, we ascertained a consanguineous Pakistani family with ARMR in four living individuals from three branches of the family, plus an additional affected individual later identified as a phenocopy. Retinitis pigmentosa was present in affected individuals, but no other features suggestive of a syndromic form of MR were found. We used Affymetrix 500K microarrays to perform homozygosity mapping and identified a homozygous and haploidentical region of 11.2 Mb on chromosome 4p15.33-p15.2. Linkage analysis across this region produced a maximum two-point LOD score of 3.59. We sequenced genes within the critical region and identified a homozygous splice-site mutation segregating in the family, within a coiled-coil and C2 domain-containing gene, CC2D2A. This mutation leads to the skipping of exon 19, resulting in a frameshift and a truncated protein lacking the C2 domain. Conservation analysis for CC2D2A suggests a functional domain near the C terminus as well as the C2 domain. Preliminary functional studies of CC2D2A suggest a possible role in Ca(2+)-dependent signal transduction. Identifying the function of CC2D2A, and a possible common pathway with CC2D1A, in correct neuronal development and functioning may help identify possible therapeutic targets for MR. PMID- 18387596 TI - Retrograde projections to a discrete apneic site in the midline medulla oblongata of the rat. AB - We have recently described a discrete region of the medullary raphe nuclei, termed the midline apneic site (MAS) that produces profound apnea upon chemical stimulation. The aim of the present study was to identify brain regions that innervate the MAS. The MAS was functionally identified and then the retrograde tracer, cholera toxin B subunit was injected in Sprague Dawley rats (n = 5). The MAS received projections of varying intensity from a number of brain regions previously associated with thermoregulation, fear, nociception, cardiovascular regulation and respiration. These include: the medial preoptic nucleus; median and lateral preoptic area; medial division of the bed nucleus of stria terminalis; paraventricular nucleus; central nucleus of the amygdala; dorsal hypothalamic area/dorsomedial hypothalamus; lateral hypothalamic area; lateral, ventrolateral and dorsomedial divisions of the periaqueductal grey; dorsal raphe nuclei; parabrachial nuclei; Kolliker-Fuse nucleus; intertrigeminal region; rostral ventrolateral medulla; lateral parafacial region; and the ventral respiratory group. The intermingling of functionally distinct cell groups in the raphe probably explains the large diversity of projections as found in other tracing studies of the raphe although the possibility remains that the MAS may integrate signals from multiple sites. The connection between the intertrigeminal region of the pons and the MAS indicates the possibility that this pathway participates in airway protective reflexes such as the Hering-Breuer and diving reflexes. One previously undescribed region that we have termed the lateral parafacial region was consistently labeled. The role of this region is yet to be determined. PMID- 18387597 TI - TAT-PAX6 protein transduction in neural progenitor cells: a novel approach for generation of dopaminergic neurones in vitro. AB - Neural stem cells (NSCs) have the potential to be used for the treatment of Parkinson's disease (PD), as they can be expanded, manipulated and differentiated in vitro to generate dopaminergic neurones which are suitable for transplantation. Since NSCs have a tendency to follow an astrocytic lineage after differentiation in vitro, researchers are investigating ways to induce a neuronal phenotype in these cells. In this study, the human immunodeficiency virus 1 (HIV 1) transactivator of transcription (TAT) protein transduction domain (PTD) system was used in an attempt to promote neuronal differentiation in rodent NSCs. A fusion protein that incorporated both the TAT PTD and the Pax6 protein (a determinant of neurogenesis) was created and added to the differentiation phase of embryonic day (E) 12 rat ventral mesencephalic (VM) neurosphere cultures. Subsequently, application of dopaminergic growth factors (GFs) was used in an attempt to induce the newly-generated neuronal progenitors to adopt a dopaminergic phenotype. In addition, a technique involving the differentiation of intact neurospheres (instead of the differentiation of neurosphere-derived dissociated cells) was investigated for its ability to promote dopaminergic neurogenesis. Immunocytochemical analysis of the differentiated neurosphere cultures indicated that both of these techniques had a significant effect on the emergence of dopaminergic neurones. Moreover, upon combination of these techniques, a further increase in dopaminergic neuronal generation was observed. Based on the findings of the present study, it is clear that NSCs are greatly influenced by their environment and that optimised in vitro conditions can support the potential of these cells to differentiate into dopaminergic neurones. PMID- 18387595 TI - On the replication of genetic associations: timing can be everything! AB - The failure of researchers to replicate genetic-association findings is most commonly attributed to insufficient statistical power, population stratification, or various forms of between-study heterogeneity or environmental influences.(1) Here, we illustrate another potential cause for nonreplications that has so far not received much attention in the literature. We illustrate that the strength of a genetic effect can vary by age, causing "age-varying associations." If not taken into account during the design and the analysis of a study, age-varying genetic associations can cause nonreplication. By using the 100K SNP scan of the Framingham Heart Study, we identified an age-varying association between a SNP in ROBO1 and obesity and hypothesized an age-gene interaction. This finding was followed up in eight independent samples comprising 13,584 individuals. The association was replicated in five of the eight studies, showing an age-dependent relationship (one-sided combined p = 3.92 x 10(-9), combined p value from pediatric cohorts = 2.21 x 10(-8), combined p value from adult cohorts = 0.00422). Furthermore, this study illustrates that it is difficult for cross sectional study designs to detect age-varying associations. If the specifics of age- or time-varying genetic effects are not considered in the selection of both the follow-up samples and in the statistical analysis, important genetic associations may be missed. PMID- 18387598 TI - Neurogenesis and neuronal commitment following ischemia in a new mouse model for neonatal stroke. AB - Stroke in the neonatal brain is an important cause of neurologic morbidity. To characterize the dynamics of neural progenitor cell proliferation and maturation after survival delays in the neonatal brain following ischemia, we utilized unilateral carotid ligation alone to produce infarcts in postnatal day 12 CD1 mice. We investigated the neurogenesis derived from the sub-ventricular zone and the sub-granular zone of the dentate gyrus subsequent to injury. Newly produced cells were labeled by bromodeoxyuridine at approximately 1 week (P18-20) after the insult by 5 i.p. injections (each 50 mg/kg). Subsequent migration and differentiation of the newborn cells was investigated at postnatal day 40 by immunohistochemistry for molecular neuronal and glial cell-lineage markers and BrdU incorporation. Cresyl violet stain demonstrated massive loss of neurons in the ipsilateral septal hippocampus in the CA3 and CA1 regions associated with atrophy. Total counts of new cells were significantly lowered not only in the ipsilateral injured but also the contralateral uninjured hippocampi and correlated with the lesion induced atrophy. Bilateral percent neuronal commitments in the dentate gyri however, were not significantly different from control. New cell densities in the neocortex and striatum increased bilaterally after neonatal stroke. The predominantly non-neuronal commitment of the SVZ derived new cells was similar to the percentage of non-neuronal commitment in controls. In conclusion, neurogenesis occurring at 1 week after neonatal ischemia in the model maintained cell-lineage commitment patterns similar to sham controls. However, the total number of hippocampal SGZ-derived new neurons was reduced bilaterally; in contrast, the SVZ-derived neurogenesis was amplified. PMID- 18387599 TI - Dose-response study of chronic alcohol induced changes in sleep patterns in rats. AB - The goal of the present study was to determine an optimum exposure regimen for alterations in sleep induced by chronic alcohol treatments in rats. We used two different exposure routes (alcohol in water and alcohol in liquid diet at two different doses in each regimen (6% and 12% alcohol in water and 3% and 6% alcohol in liquid diet)). All treatments were for 6 weeks. We found the effects of the 6% and 12% in water and 3% in liquid diet to be very similar; all three produced increases in slow-wave sleep (SWS) only in the dark period with no changes in rapid-eye-movement sleep (REMS). On the other hand 6% alcohol in liquid diet caused much more dramatic changes, with alterations in both SWS and REMS in both the dark and light periods. These animals spent less time in SWS and REMS during the light period but more time in SWS and REMS in the dark period. Additionally, the variation of slow-wave amplitude (SWA) across day and night in this group of alcoholic animals is blunted with the loss of the peak of SWA at the beginning of light onset compared to the other groups. We conclude that future alcohol treatment regimens used to investigate the effects of alcohol on sleep in adult rats should use an exposure protocol of at least 6 weeks with 6% alcohol in liquid diet. PMID- 18387600 TI - Neuregulin blocks synaptic strengthening after epileptiform activity in the rat hippocampus. AB - Synaptic strengthening produced by epileptiform activity may contribute to seizure progression and cognitive impairment in epilepsy. Agents that limit this form of plasticity may have therapeutic benefit. Neuregulin is an endogenous growth factor that is released at synapses in an activity dependent manner and can suppress long term potentiation (LTP). Alterations in neuregulin signaling have been associated with schizophrenia. A role for neuregulin in epilepsy has not been explored. We used field potential recordings to examine the role of neuregulin in regulating synaptic strengthening following epileptiform activity in hippocampal slices. Neuregulin had no effect on basal synaptic transmission, isolated NMDA field potentials or GABAergic inhibition on CA1 pyramidal neurons. However, it reversed LTP at CA1 synapses. Brief exposure to 10 mM potassium chloride produced epileptiform bursting and potentiation of CA1 synapses and suppressed the subsequent induction of LTP. Neuregulin reversed high K(+)-induced synaptic strengthening, enabling LTP induction after neuregulin washout. In this manner neuregulin preserved the dynamic range of synaptic responses and plasticity after epileptiform activity. These results indicate that LTP and high K(+)-induced synaptic strengthening share a common neuregulin-sensitive mechanism. By opposing synaptic strengthening caused by epileptiform activity, we suggest that neuregulin may reduce the generation and spread of seizures as well as memory deficits associated with epilepsy. PMID- 18387601 TI - A developmental ERP study of verbal and non-verbal semantic processing. AB - To clarify how different the processing of verbal information is from the processing of meaningful non-verbal information, the present study characterized the developmental changes in neural responses to words and environmental sounds from pre-adolescence (7-9 years) through adolescence (12-14 years) to adulthood (18-25 years). Children and adults' behavioral and electrophysiological responses (the N400 effect of event-related potentials) were compared during the processing of words and environmental sounds presented in semantically matching and mismatching picture contexts. Behavioral accuracy of picture-sound matching improved until adulthood, while reaction time measures leveled out by age 12. No major electrophysiological changes in the N400 effect were observed between pre adolescence and adolescence. When compared to adults, children demonstrated significant maturational changes including longer latencies and larger amplitudes of the N400 effect. Interestingly, these developmental differences were driven by stimulus type: the Environmental Sound N400 effect decreased in latency from adolescence to adulthood, while no age effects were observed in response to Words. Thus, while the semantic processing of single words is well established by 7 years of age, the processing of environmental sounds continues to improve throughout development. PMID- 18387602 TI - Beta-catenin is required for the establishment of vegetal embryonic fates in the nemertean, Cerebratulus lacteus. AB - Downstream components of the canonical Wnt signaling pathway that result in the nuclear localization of beta-catenin are involved in diverse developmental processes including the formation of the mesendoderm, the regulation of axial properties and asymmetric cell divisions in a wide array of metazoans. The nemertean worm, Cerebratulus lacteus, represents a member of the understudied lophotrochozoan clade that exhibits a highly stereotyped spiral cleavage program in which ectodermal, endodermal, and mesodermal origins are known from intracellular fate mapping studies. Here, the embryonic distribution of beta catenin protein was studied using injection of synthetic mRNA, encoding GFP tagged beta-catenin, into fertilized eggs. During the early cleavage stages beta catenin was destabilized/degraded in animal hemisphere blastomeres and became localized to the nuclei of the four vegetal-most cells at the 64-cell stage, which give rise to definitive larval and adult endoderm. Functional assays indicate that beta-catenin plays a key role in the development of the endoderm. Morpholino knockdown of endogenous beta-catenin, as confirmed by Western analysis, resulted in the failure to gastrulate, absence of the gut and an animalized phenotype in the resulting larvae, including the formation of ectopic (anterior) apical organ tissue with elongated apical tuft cilia and no indications of dorsoventral polarity. Similarly, over-expression of the cytoplasmic domain of cadherin or a beta-catenin-engrailed repressor fusion construct prevented endoderm formation and generated the same animalized phenotype. Injections of mRNA encoding either a stabilized, constitutively activated form of beta-catenin or a dominant negative form of GSK3-beta converted all or nearly all cells into endodermal fates expressing gut-specific esterase. Thus, beta-catenin appears to be both necessary and sufficient to promote endoderm formation in C. lacteus, consistent with its role in endoderm and endomesoderm formation in anthozoan cnidarians, ascidians, and echinoderms. Consistent with the results of other studies, beta-catenin may be viewed as playing a role in the development of posterior/vegetal larval fates (i.e., endoderm) in C. lacteus. However, unlike the case found in polychaete annelid and soil nematode embryos, there is no evidence for a role of beta-catenin in regulating cell fates and asymmetric cell divisions along the entire anterior posterior axis. PMID- 18387604 TI - Therapeutic effect of magnesium lithospermate B on neointimal formation after balloon-induced vascular injury. AB - Vascular smooth muscle cell (VSMC) proliferation and migration in response to platelet-derived growth factor (PDGF) play an important role in the development of atherosclerosis and restenosis. Recent evidence indicates that PDGF increases intracellular levels of reactive oxygen species in VSMCs and that both PDGF induced VSMC proliferation and migration are reactive oxygen species-dependent. Danshen is a representative oriental medicine used for the treatment of vascular disease. Previously, we reported that magnesium lithospermate B, an active component of Danshen, is a potent antioxidant. Thus we investigated the therapeutic potential of magnesium lithospermate B in neointimal formation after carotid artery injury in rats along with its effects on the PDGF signaling pathway for stimulating VSMC proliferation and migration in vitro. PDGF is dimeric glycoprotein composed of two A or two B chains. In this study, we used PDGF-BB, which is one of the isoforms of PDGF (i.e., PDGF-AA, PDGF-BB, and PDGF AB). Our results demonstrated that magnesium lithospermate B directly scavenged reactive oxygen species in a xanthine/xanthine oxidase system and reduced PDGF-BB induced intracellular reactive oxygen species generation in VSMCs. In a rat carotid artery balloon injury model, magnesium lithospermate B treatment (10 mg/kg/day, i.p) showed a significant effect on the prevention of neointimal formation compared with vehicle treatment. In cultured VSMCs, magnesium lithospermate B significantly attenuated PDGF-BB-induced cell proliferation and migration as measured by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-tetrazolium bromide (MTT) assay and transwell migration assays, respectively. Further, magnesium lithospermate B inhibited PDGF-BB-induced phosphorylation of phospatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathways by scavenging reactive oxygen species. Together, these data indicated that magnesium lithospermate B, a potent reactive oxygen species scavenger, prevented both injury-induced neointimal formation in vivo and PDGF-BB-induced VSMC proliferation and migration in vitro, suggesting that magnesium lithospermate B may be a promising agent to prevent atherosclerosis and restenosis following angioplasty. PMID- 18387605 TI - The action of extracellular NAD+ in the liver of healthy and tumor-bearing rats: model analysis of the tumor-induced modified response. AB - The chronic inflammatory state induced by cancer is expected to affect the actions of extracellular NAD(+) in the liver because these are largely mediated by eicosanoids. Under this assumption the present work was planned to investigate the influence of the Walker-256 tumor on the action of extracellular NAD(+) on metabolism and hemodynamics in the perfused rat liver. The experiments were done with livers from healthy and tumor-bearing rats with measurements of gluconeogenesis from lactate, pyruvate production, oxygen consumption and portal pressure. A model describing the biphasic effects of NAD(+) was proposed as an auxiliary worktool for interpretation. The Walker-256 tumor modified the responses of metabolism to extracellular NAD(+) by delaying the peak of maximal responses and by prolonging the inhibitory effects. The transient increase in portal perfusion pressure caused by NAD(+) was enhanced and delayed. The model was constructed assuming the mediation of a down-regulator (inhibition), an up regulator (stimulation) and receptor dessensitization. Analysis suggested that the productions of both the down- and up-regulators were substantially increased and delayed in time in the tumor-bearing condition. Since the regulators are probably eicosanoids, this analysis is consistent with the increased capacity of producing these agents in the chronic inflammatory state induced by cancer. PMID- 18387603 TI - The As and Ds of stress: metabolic, morphological and behavioral consequences. AB - Unlike responses to acute stressful events that are protective and adaptive in nature, chronic stress elicits neurochemical, neuroanatomical and cellular changes that may have deleterious consequences upon higher brain functioning. For example, while exposure to acute stress facilitates memory formation and consolidation, chronic stress or chronic exposure to stress levels of glucocorticoids impairs cognitive performance. Chronic stress or glucocorticoid exposure, as well as impairments in hypothalamic-pituitary-adrenal (HPA) axis function are proposed to participate in the etiology and progression of neurological disorders such as depressive illness, anxiety disorders and post traumatic stress disorder (PTSD). HPA axis dysfunction, impaired stress responses and elevated basal levels of glucocorticoids are also hallmark features of experimental models of type 1 and type 2 diabetes, as well as diabetic subjects in poor glycemic control. Such results suggest that stress and glucocorticoids contribute to the neurological complications observed in diabetes patients. Interestingly, many of the hyperglycemia mediated changes in the brain are similar to those observed in depressive illness patients and in experimental models of chronic stress. Such results suggest that common mechanisms may be involved in the development of the neurological complications associated with Anxiety, Depressive illness and Diabetes: the As and Ds of stress. The aim of the current review will be to discuss the mechanisms through which limbic structures such as the hippocampus and amygdala respond and adapt to the deleterious consequences of chronic stress and hyperglycemia. PMID- 18387607 TI - Recombinant VSV G proteins reveal a novel raft-dependent endocytic pathway in resorbing osteoclasts. AB - Transcytotic membrane flow delivers degraded bone fragments from the ruffled border to the functional secretory domain, FSD, in bone resorbing osteoclasts. Here we show that there is also a FSD-to-ruffled border trafficking pathway that compensates for the membrane loss during the matrix uptake process and that rafts are essential for this ruffled border-targeted endosomal pathway. Replacing the cytoplasmic tail of the vesicular stomatitis virus G protein with that of CD4 resulted in partial insolubility in Triton X-100 and retargeting from the peripheral non-bone facing plasma membrane to the FSD. Recombinant G proteins were subsequently endosytosed and delivered from the FSD to the peripheral fusion zone of the ruffled border, which were both rich in lipid rafts as suggested by viral protein transport analysis and visualizing the rafts with fluorescent recombinant cholera toxin. Cholesterol depletion by methyl-beta-cyclodextrin impaired the ruffled border-targeted vesicle trafficking pathway and inhibited bone resorption dose-dependently as quantified by measuring the CTX and TRACP 5b secreted to the culture medium and by measuring the resorbed area visualized with a bi-phasic labeling method using sulpho-NHS-biotin and WGA-lectin. Thus, rafts are vital for membrane recycling from the FSD to the late endosomal/lysosomal ruffled border and bone resorption. PMID- 18387606 TI - PGAM5 tethers a ternary complex containing Keap1 and Nrf2 to mitochondria. AB - Eukaryote cells balance production of reactive oxygen species (ROS) with levels of anti-oxidant enzyme activity to maintain cellular redox homeostasis. Mitochondria are a major source of ROS, while many anti-oxidant genes are regulated by the Nrf2 transcription factor. Keap1, a redox-regulated substrate adaptor for a cullin-based ubiquitin ligase, targets Nrf2 for proteosome-mediated degradation and represses Nrf2-dependent gene expression. We have previously identified a member of the phosphoglycerate mutase family, PGAM5, as a Keap1 binding protein. In this report, we demonstrate that PGAM5 is targeted to the outer membrane of mitochondria by an N-terminal mitochondrial-localization sequence. Furthermore, we provide evidence that PGAM5 forms a ternary complex containing both Keap1 and Nrf2, in which the dimeric Keap1 protein simultaneously binds both PGAM5 and Nrf2 through their conserved E(S/T)GE motifs. Knockdown of either Keap1 or PGAM5 activates Nrf2-dependent gene expression. We suggest that this ternary complex provides a molecular framework for understanding how nuclear anti-oxidant gene expression is regulated in response to changes in mitochondrial function(s). PMID- 18387608 TI - NK and CD4+ T cell changes in blood after seizures in temporal lobe epilepsy. AB - Immunological phenomena may affect the course of focal epilepsy. We analyzed prospectively the pre- and postictal distribution of leukocyte subsets in epileptic patients. METHODS: Twenty-two patients (age 36.6+/-10.8 years, 50% men) with temporal lobe epilepsy were included. Distribution of leukocyte subsets and serum levels of epinephrine were measured in peripheral blood immediately and 24 h after seizures and compared to baseline values. RESULTS: In the immediate postictal state (10+/-6 min), we observed a significant relative increase of total leukocytes (42%, p=0.0004), neutrophil leukocytes (55%, p=0.0007), total lymphocytes (45%, p=0.0019), natural killer (NK) cells (104%, p=0.0017), and epinephrine (454%, p=0.0014). CD4(+) T cells decreased by 13% (p=0.0113). These postictal changes remained significant considering only complex partial seizures (n=17). The alterations were more pronounced in patients with hippocampal sclerosis. Treatment with valproic acid (VPA) was accompanied by a greater postictal decrease of CD4(+) T cells (25% compared to 5% in patients without VPA, p=0.041) while treatment with levetiracetam (LEV) correlated with a low postictal increase of NK-like T cells (4% versus 41%, p=0.016). Twenty-four hours after the seizures the alterations had resolved. CONCLUSION: Profound postictal changes in the immune cell composition of the peripheral blood may have been mediated by epinephrine release. The greater immune response in patients with mesial temporal lobe epilepsy due to hippocampal sclerosis may reflect a close relationship between mesial temporal structures and the sympathetic nerve system. VPA and LEV may have an impact on seizure induced immunological changes. PMID- 18387609 TI - Trypanosoma cruzi: effects of social stress in Calomys callosus a natural reservoir of infection. AB - Social environment can represent a major source of stress affecting cortisol and/or corticosterone levels, thereby altering the immune response. We have investigated the effects of social isolation on the development of Trypanosoma cruzi infection in female Calomys callosus, a natural reservoir of this protozoan parasite. Animals were divided in groups of five animals each. The animals of one group were kept together in a single cage. In a second group, four females were kept together in a cage with one male. In the final group, five individuals were kept isolated in private cages. The isolated animals showed body weight reduction, decreased numbers of peritoneal macrophages, lower global leucocytes counts, smaller lytic antibody percentage and a significantly higher level of blood parasites compared to the other animals. Their behavior was also altered. They were more aggressive than grouped females, or females exposed to the presence of a male. These results suggest that isolation creates a distinct social behavior in which immunity is impaired and pathogenesis is enhanced. PMID- 18387611 TI - Day length and estradiol affect same-sex affiliative behavior in the female meadow vole. AB - Non-sexual social bonding between adult mammals is poorly understood, despite its importance in many species. Female meadow voles are territorial and nest alone in long summer day lengths when circulating estradiol concentrations are high, but cohabit in groups in short winter photoperiods when estradiol secretion is low. The influence of day length and estradiol on same-sex huddling behavior was assessed in adult female pairs housed together in long day lengths (LDs) or short day lengths (SDs) from weaning. The behavior of intact, ovariectomized, and estradiol-treated ovariectomized females from each photoperiod was assessed during 3 h partner-preference tests. Intact SD voles, unlike intact LD voles, spent the majority of the test in proximity to their cage-mates. Estradiol treatment of SD voles significantly reduced time spent huddling with the partner. Neither ovariectomy nor estradiol treatment significantly affected the amount of time LD females spent in contact with their partners. Low estradiol availability is therefore a necessary but not sufficient condition for maintenance of high levels of huddling. These results establish that ovarian hormones interact with photoperiod to affect same-sex social behavior. PMID- 18387612 TI - Perinatal factors associated with the neurologic impairment of children born preterm. AB - OBJECTIVE: To determine the perinatal risk factors of long-term neurologic impairment for preterm infants. METHODS: A case-control study was conducted with 60 neurologically impaired and 60 healthy children, all born prematurely. RESULTS: There was no relation between neurologic impairment and maternal pregnancy complications or prenatal steroid administration, bacteriologic content of cervical smear, fetal presentation, fetal heart rate, or mode of delivery. Cerebral palsy was associated with early neurologic signs, perinatal asphyxia, neonatal septicemia, abnormal brain ultrasound findings, prolonged interval between rupture of membranes and delivery, and multiple placental lesions. Children with minimal cerebral dysfunction were more frequently first born. Multiple placental lesions, neonatal septicemia, abnormal brain ultrasound findings, and perinatal asphyxia were independently correlated with long-term neurologic impairment. CONCLUSION: Perinatal infection, perinatal asphyxia, and abnormal brain ultrasound findings seem to be risk factors for cerebral palsy whereas primigravidity seems to be correlated with minimal cerebral dysfunction. PMID- 18387613 TI - Priorities for women's health from the Global Burden of Disease study. AB - Women's health is increasingly recognized as a global health priority. It is essential to address not only sexual and reproductive health, but also other health issues that occur throughout life. This paper uses the 2005 estimates of morbidity and mortality from the WHO Global Burden of Disease study and a review of the literature to identify key areas of concern for women by age group and world region. The leading causes of death in women aged between 15 and 44 years include infectious diseases such as HIV/AIDS, tuberculosis, maternal health conditions, and injuries. The leading causes of death in women aged 45 years and older include cardiovascular diseases, chronic obstructive pulmonary disease, and other noncommunicable conditions. Neuropsychiatric and sensory disorders are a major cause of disability for both age groups. An understanding of the key regional and age-specific priorities for women's health will facilitate the development of appropriate interventions and policies to reduce disease burden. PMID- 18387610 TI - Androgen regulation of axon growth and neurite extension in motoneurons. AB - Androgens act on the CNS to affect motor function through interaction with a widespread distribution of intracellular androgen receptors (AR). This review highlights our work on androgens and process outgrowth in motoneurons, both in vitro and in vivo. The actions of androgens on motoneurons involve the generation of novel neuronal interactions that are mediated by the induction of androgen dependent neurite or axonal outgrowth. Here, we summarize the experimental evidence for the androgenic regulation of the extension and regeneration of motoneuron neurites in vitro using cultured immortalized motoneurons, and axons in vivo using the hamster facial nerve crush paradigm. We place particular emphasis on the relevance of these effects to SBMA and peripheral nerve injuries. PMID- 18387614 TI - Cross-bridge induced force enhancement? AB - When a muscle is stretched while activated, its steady-state isometric force following stretch is greater than the corresponding purely isometric force. This so-called residual force enhancement (RFE) has been observed for half a century, yet its mechanism remains unknown. Recent experiments suggest that RFE is not caused by non-uniformities in sarcomere lengths, as had been assumed for a long time, and cannot be explained primarily with increases in passive force, but is directly related to the kinetics of the cross-bridge cycle. Specifically, it has been suggested that stretching an attached cross-bridge increases its dwell time and duty ratio; therefore, the proportion of attached cross-bridges in a muscle would be increased by stretch, thereby causing RFE. A three bead laser trap setup was used for testing single cross-bridge (myosin II) interactions with actin. Upon attachment of a cross-bridge, a stretch or shortening of the cross-bridge was applied with a force of about 1.0 pN. The hypothesis that stretching a single cross-bridge increases its dwell time and duty ratio was rejected. However, stretching caused an increase in the average steady-state force per cross-bridge (3.4+/-0.4 pN; n=433) compared to shortening (1.9+/-0.3 pN; n=689). Therefore, based on the results of this study, RFE cannot be explained by an increased duty ratio and the associated increase in proportion of attached cross-bridges, but might be associated with an increased force per cross-bridge. PMID- 18387615 TI - Estimation of the effective static moment arms of the tendons in the index finger extensor mechanism. AB - A novel technique to estimate the contribution of finger extensor tendons to joint moment generation was proposed. Effective static moment arms (ESMAs), which represent the net effects of the tendon force on joint moments in static finger postures, were estimated for the 4 degrees of freedom (DOFs) in the index finger. Specifically, the ESMAs for the five tendons contributing to the finger extensor apparatus were estimated by directly correlating the applied tendon force to the measured resultant joint moments in cadaveric hand specimens. Repeated measures analysis of variance revealed that the finger posture, specifically interphalangeal joint angles, had significant effects on the measured ESMA values in 7 out of 20 conditions (four DOFs for each of the five muscles). Extensor digitorum communis and extensor indicis proprius tendons were found to have greater MCP ESMA values when IP joints are flexed, whereas abduction ESMAs of all muscles except extensor digitorum profundus were mainly affected by MCP flexion. The ESMAs were generally smaller than the moment arms estimated in previous studies that employed kinematic measurement techniques. Tendon force distribution within the extensor hood and dissipation into adjacent structures are believed to contribute to the joint moment reductions, which result in smaller ESMA values. PMID- 18387616 TI - Experimental probing and modeling of key sorbent-solute interactions of norephedrine enantiomers with polysaccharide-based chiral stationary phases. AB - The key interactions of a chiral solute, norephedrine or 2-amino-1-phenyl-1 propanol (PPA), with three commercially important polysaccharide-based chiral stationary phases, amylose Tris(3,5-dimethylphenylcarbamate) (ADMPC), amylose Tris((S)-alpha-methylbenzylcarbamate) (ASMBC) and cellulose Tris(3,5 dimethylphenylcarbamate) (CDMPC) are studied in detail using different experimental techniques and molecular simulations. The HPLC retention factors of the enantiomers of PPA in n-hexane/2-propanol (90/10, v/v) at 298 K vary significantly with these sorbents. The enantioselectivities of -PPA versus +PPA are 2.4, 1.0, and 0.8 (reversal in the elution order), respectively. The observed changes in the wavenumbers and the intensities of the amide bands of these polymers in the attenuated total reflection-infrared spectroscopy (ATR-IR) spectra upon absorption of each enantiomer are different. The IR wavenumbers, and the H-bonding interaction energies of the polymer side chains with each enantiomer (polymer-solute) in four different binding configurations are estimated and ranked using the density functional theory (the DFT/B3LYP/6 311+g(d,p) level of theory). X-ray diffraction (XRD) results show that the polymer crystallinity increases significantly upon absorption of each enantiomer. The helical pitches and the inter-rod packing for these polymers are inferred from the XRD results and incorporated into the molecular dynamics (MD) simulations. The elution orders predicted for the enantiomers of PPA using MD simulations of the polymer-PPA binary systems are consistent with the chromatography results. The enantioselectivity observed in ADMPC is hypothesized to be due to having three simultaneous interactions (two H-bond and one pi-pi) of the polymer with -PPA versus two interactions (one H-bond and one pi-pi) with +PPA. PMID- 18387617 TI - Direct deposition of gold nanoparticles onto polymer beads and glucose oxidation with H2O2. AB - Gold nanoparticles (Au NPs) were deposited directly from aqueous solution of diethylenediaminegold(III) complex onto polymer beads commercially available, such as poly(methyl methacrylate) (PMMA), polystyrene (PS), and polyaniline (PANI) without surface modification. The dropwise addition of NaBH4 to reduce Au(III) was found to be very effective to obtain small Au0 NPs with a narrow size distribution except for PANI. The catalytic performance of Au NPs deposited on polymer beads for H2O2 decomposition and glucose oxidation with H2O2 were more significantly affected by the kinds of polymer supports than by the size of Au NPs. The equimolar oxidation of glucose with H2O2 could be operated by controlling the decomposition rate of H2O2 over Au/PMMA. PMID- 18387618 TI - Modeling the surface charge evolution of spherical nanoparticles by considering dielectric discontinuity effects at the solid/electrolyte solution interface. AB - It is well known that the electrostatic repulsions between charges on neighboring sites decrease the effective charge at the surface of a charged nanoparticle (NP). However, the situation is more complex close to a dielectric discontinuity, since charged sites are interacting not only with their neighbors but also with their own image charges and the image charges of all neighbors. Titrating site positions, solution ionic concentration, dielectric discontinuity effects, and surface charge variations with pH are investigated here using a grand canonical Monte Carlo method. A Tanford and Kirkwood approach is used to calculate the interaction potentials between the discrete charged sites. Homogeneous, heterogeneous, and patch site distributions are considered to reproduce the various titrating site distributions at the solid/solution interface of spherical NPs. By considering Coulomb, salt, and image charges effects, results show that for different ionic concentrations, modifications of the dielectric constant of NPs having homogeneous and heterogeneous site distributions have little effect on their charging process. Thus, the reaction field, due to the presence of image charges, fully counterbalances the Coulomb interactions. This is not the case for patch distributions, where Coulomb interactions are not completely counterbalanced by the reaction field. Application of the present model to pyrogenic silica is also performed and comparison is made with published experimental data of titration curves at various ionic concentrations. PMID- 18387619 TI - Synergistic effects in mixtures of oppositely charged surfactants as calculated from the Poisson-Boltzmann theory: a comparison between theoretical predictions and experiments. AB - Critical micelle concentrations in mixtures of an anionic surfactant and a cationic amphiphilic drug have been investigated using a model-independent procedure to quantify observed synergistic effects. Experimental results were compared with a theory based on the Poisson-Boltzmann mean field approximation of a charged interface with a diffuse layer of counterions. Explicit expressions for the activity coefficients from which the critical micelle concentration can be calculated and quantitatively predicted have been derived and excellent agreement between experimental data and theory was obtained. As a result, we demonstrate that it is possible to rationalize and predict the magnitude of synergism in mixtures of oppositely charged surfactants in the presence of added salt. PMID- 18387620 TI - Thermodynamics of naphthalene sorption to organoclays: role of surfactant packing density. AB - Temperature dependence of naphthalene sorption to four organoclays with different surfactant (CTMA+) packing densities was examined. The results showed that both DeltaH o and DeltaS o increase generally with CTMA+ packing density. For organoclays with a low CTMA(+) packing density, the sorption process is driven by both the enthalpy term (DeltaH(o)) and the entropy term (-T DeltaS o), with values ranging from -4.7 to -7.5 kJ mol(-1) and -15.9 to -20.8 kJ mol(-1), respectively. As the CTMA+ packing density increases, the sorption process is driven by the entropy term (from -29.2 to -65.0 kJ mol(-1)) while it is opposed by the enthalpy term (from 7.9 to 40.5 kJ mol(-1)). These results indicate that the enthalpy demand for cavity formation within the surfactant aggregates and the mixing entropy of solute with surfactant aggregates both increase with the surfactant packing density. This means that the surfactant aggregates will form various organic phases as their packing density varies. Controlling the surfactant aggregates within an intermediate packing density range can improve the sorption capacities of the organoclays. PMID- 18387621 TI - One-step synthesis and functionalization of hydroxyl-decorated magnetite nanoparticles. AB - Magnetite nanoparticles covered by a layer of omega-hydroxycarboxylic acid were synthesized in one step by high-temperature decomposition of iron(III) omega hydroxycarboxylates in tri- and tetra-ethylene glycol. The nanoparticles were characterized by TEM, XRD, IR, XPS and NMR techniques in order to show that they comprise a crystalline magnetite core and actually bear on the outer surface terminal hydroxy groups. The latter ones are convenient "handles" for further functionalization as opposed to the chemically-inert aliphatic chains which cover conventionally synthesized nanoparticles. This was shown by several examples in which the hydroxy groups on the nanoparticle surface were easily transformed in other functional groups or reacted with other molecules. For instance, the hydroxyl-decorated nanoparticles were made water soluble by esterification with a PEGylated acetic acid. The reactive behavior of the surfactant monolayer was monitored by degrading the nanoparticles with aqueous acid and isolating the surfactant for NMR characterization. In general, the reactivity of the terminal hydroxyl groups on the nanoparticle surface parallels that observed in the free surfactants. The reported hydroxyl-decorated magnetite nanoparticles can be thus considered as pro-functional nanoparticles, i.e., a convenient starting material to functionalized magnetic nanoparticles. PMID- 18387622 TI - Aluminum hydroxide microstructural units in gelled media aged, or nonaged, with alcohol and water. AB - Aluminum hydroxide gelled media dispersed and then aged in alcohol (ES) or in water (WS) were prepared with a solid phase concentration equal to 0.45% v/v. The media were characterized through rheological measurements and detailed analysis. Xerogels were obtained through a designed drying process at 8 degrees C under vacuum for 24 h, followed by a heating treatment at 105 degrees C and a pressure of 10(-3) Pa for 2 h. The xerogel microstructure was carefully investigated using N2 adsorption-desorption methods at 78 K. Rheology data on ES and WS colloidal dispersions were compared with data computed from adsorption isotherms of corresponding xerogels, in order to infer information about particles organization in the dispersions. On this ground, a model for microstructural units dispersed in the liquid phase has been derived. The beneficial action of alcohol in giving xerogel powders with higher specific surface area and higher porosity has been explained on the basis that alcohol produces changes in the corresponding dispersed microstructural units. The effect of the aging time in alcohol and in water was evaluated, and the evolution of all microstructural parameters was described. PMID- 18387623 TI - Fabrication of highly insulating tethered bilayer lipid membrane using yeast cell membrane fractions for measuring ion channel activity. AB - A tethered bilayer lipid membrane (tBLM) was fabricated on a gold electrode using 1,2-dipalmitoyl-sn-glycero-phosphothioethanol as a tethering lipid and the membrane fractions of Saccharomyces pombe yeast cells to deposit the upper leaflet. The membrane fractions were characterized using transmission electron microscopy and dynamic light scattering and found to be similar in size to small unilamellar vesicles of synthetic lipids. The dynamics of membrane-fraction deposition and rupture on the tethering-lipid layer were measured using quartz crystal microgravimetry. The electrochemical properties of the resulting tBLM were characterized using electrical impedance spectroscopy and cyclic voltammetry. The tBLM's electrical resistance was greater than 1 MOmegacm(2), suggesting a defect-free membrane. The suitability of tBLM produced using membrane fractions for measuring ion-channel activities was shown by a decrease in membrane resistance from 1.6 to 0.43 MOmegacm(2) following addition of gramicidin. The use of membrane fractions to form high-quality tBLM on gold electrodes suggests a new approach to characterize membrane proteins, in which the upper leaflet of the tBLM is deposited, and overexpressed membrane proteins are incorporated, in a single step. This approach would be especially useful for proteins whose activity is lost or altered during extraction, purification, and reconstitution, or whose activities are strongly influenced by the lipid composition of the bilayer. PMID- 18387624 TI - Side-chain-controlled H- and J-aggregation of amphiphilic porphyrins in CTAB micelles. AB - The aggregation behavior of a series of amphiphilic 4-hydroxyphenyl porphyrins with one (P1), two (P2) and three (P3) hexadecyl side chains in cetyltrimethylammonium bromide (CTAB) micelles has been studied by means of UV vis and fluorescence spectra. It was found that the number of hexadecyl side chains not only controls the H- and J-aggregation of the porphyrins in CTAB micelles, but also influences the aggregation concentration and tendency. With increasing porphyrin concentration, P1 and P2 form H-aggregates in CTAB micelles, while P3 forms J-aggregates. Porphyrins with more hexadecyl side chains tend to form aggregates more easily and at lower concentrations in CTAB micellar solutions. PMID- 18387625 TI - Nanoporosities and catalytic activities of Pd-tailored single wall carbon nanohorns. AB - The nanoporosities and catalytic activities of Pd nanoparticles dispersed on single wall carbon nanohorns (Pd-SWCNHs) and oxidized single wall carbon nanohorns (Pd-ox-SWCNHs) were examined. A transmission electron microscopy (TEM) observation indicated that Pd nanoparticles of 2-3 nm size were highly dispersed on both the SWCNHs. X-ray photoelectron spectra and N2 adsorption isotherms at 77 K illustrated the differences in the deposition process mechanisms of the Pd SWCNHs and Pd-ox-SWCNHs; the deposition process depended on the surface functional groups. The supercritical H2 adsorption isotherms at 77 K suggested the relationships between the interaction of Pd-SWCNHs and Pd-ox-SWCNHs with H2 and the catalytic activities for a water formation reaction in a gas phase at 273 or 298 K. The catalytic activity measurement and TEM observation of the catalysts after the reactions demonstrated that the Pd-SWCNHs and Pd-ox-SWCNHs are promising catalysts. PMID- 18387626 TI - Experimental chronic wasting disease (CWD) in the ferret. AB - Chronic wasting disease (CWD), a prion disease of North American deer, elk and moose, affects both free-ranging and captive cervids. The potential host range for CWD remains uncertain. The susceptibility of the ferret to CWD was examined experimentally by administering infectious brain material by the intracerebral (IC) or oral (PO) route. Between 15 and 20 months after IC inoculation, ferrets developed neurological signs consistent with prion disease, including polyphagia, somnolence, piloerection, lordosis and ataxia. Upon first sub-passage of ferret adapted CWD, the incubation period decreased to 5 months. Spongiform change in the neuropil was most marked in the basal ganglia, thalamus, midbrain and pons. The deposition of PrP(CWD) was granular and was occasionally closely associated with, or localized within, neurons. There were no plaque-like or perivascular PrP aggregates as seen in CWD-infected cervids. In western blots, the PrP(CWD) glycoform profile resembled that of CWD in deer, typified by a dominant diglycosylated glycoform. CWD disease in ferrets followed IC but not PO inoculation, even after 31 months of observation. These findings indicate that CWD-infected ferrets share microscopical and biochemical features of CWD in cervids, but appear to be relatively resistant to oral infection by primary CWD inoculum of deer origin. PMID- 18387627 TI - Sexual dimorphism in the attachment ability of the Colorado potato beetle Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) to rough substrates. AB - Many representatives of the beetle family Chrysomelidae exhibit a distinctive sexual dimorphism in the structure of adhesive tarsal setae. The present study demonstrates the influence of surface roughness on the friction force of Leptinotarsa decemlineata males and females. The maximum friction force of individual beetles was measured on epoxy resin surfaces (smooth and with asperities ranging from 0.3 to 12.0 microm) using a centrifugal force tester. On the smooth surface, no considerable differences between males and females were found, whereas on rough surfaces, females attached significantly (up to two times) stronger than males. Clawless beetles generated lower forces than intact ones, but demonstrated similar differences between males and females. The results indicate that the female adhesive system has its main functional trait in a stronger specialisation to rough plant surfaces whereas the adhesive system of males possess a certain trade-off between attachment to rough plant surfaces during locomotion on vegetation and to the smooth surface of the female elytra, while mating. PMID- 18387628 TI - Isolation of cockroach Phe-Gly-Leu-amide allatostatins from the termite Reticulitermes flavipes and their effect on juvenile hormone synthesis. AB - Immunoreactivity to cockroach Diploptera punctata allatostatin-7 (Dippu AST-7) has been demonstrated previously in axons innervating the corpora allata of the termite Reticulitermes flavipes. This peptide and Dippu AST-11 inhibited juvenile hormone (JH) synthesis by corpora allata (CA) of brachypterous neotenic reproductives (secondary reproductives) of termites. The present study shows that R. flavipes CA are also inhibited by Dippu AST-2, AST-5, AST-8, and AST-9 at approximately the same rank order of potency as demonstrated in D. punctata. Another allatostatin from Periplaneta americana (Peram AST-12) also inhibits JH synthesis by R. flavipes CA. Sensitivity to the allatostatins is higher in glands with low rates of JH synthesis than in those with relatively high JH synthetic rates as has been demonstrated in CA from male and female secondary reproductives as well as in those from non-egg-laying and egg-laying females. The identical inhibitory effects of R. flavipes brain extract on CA from both D. punctata and R. flavipes and the isolation and identification of five cockroach allatostatins (Dippu AST-1, AST-2, AST-5, AST-8, and Peram AST-12) from termite brain extract reflect the close relationship between cockroaches and termites. PMID- 18387629 TI - Effect of storage temperature on survival and infectivity of Steinernema rarum (OLI strain) (Rhabditida: Steinernematidae). AB - Nematode strains of the entomopathogenic family Steinernematidae differ in their ability to infect insects at different temperatures. Survival and infectivity of infective juveniles (IJs) of Steinernema rarum (OLI) were studied after their storage at 23+/-2 degrees C and at 5+/-1 degrees C. Survival at 23+/-2 degrees C was always above 95%. At 5+/-1 degrees C, survival decreased at week 5, but infectivity did the same after week 2. Unlike other steinernematids, both infectivity and survival of IJs would be higher for S. rarum (OLI) when stored at 23+/-2 degrees C. PMID- 18387630 TI - Mutation that dramatically alters rat titin isoform expression and cardiomyocyte passive tension. AB - Titin is a very large alternatively spliced protein that performs multiple functions in heart and skeletal muscles. A rat strain is described with an autosomal dominant mutation that alters the isoform expression of titin. While wild type animals go through a developmental program where the 3.0 MDa N2B becomes the major isoform expressed by two to three weeks after birth (approximately 85%), the appearance of the N2B is markedly delayed in heterozygotes and never reaches more than 50% of the titin in the adult. Homozygote mutants express a giant titin of the N2BA isoform type (3.9 MDa) that persists as the primary titin species through ages of more than one and a half years. The mutation does not affect the isoform switching of troponin T, a protein that is also alternatively spliced with developmental changes. The basis for the apparently greater size of the giant titin in homozygous mutants was not determined, but the additional length was not due to inclusion of sequence from larger numbers of PEVK exons or the Novex III exon. Passive tension measurements using isolated cardiomyocytes from homozygous mutants showed that cells could be stretched to sarcomere lengths greater than 4 mum without breakage. This novel rat model should be useful for exploring the potential role of titin in the Frank Starling relationship and mechano-sensing/signaling mechanisms. PMID- 18387631 TI - NMR spectroscopy of the ligand-binding core of ionotropic glutamate receptor 2 bound to 5-substituted willardiine partial agonists. AB - Glutamate receptors mediate neuronal intercommunication in the central nervous system by coupling extracellular neurotransmitter-receptor interactions to ion channel conductivity. To gain insight into structural and dynamical factors that underlie this coupling, solution NMR experiments were performed on the bilobed ligand-binding core of glutamate receptor 2 in complexes with a set of willardiine partial agonists. These agonists are valuable for studying structure function relationships because their 5-position substituent size is correlated with ligand efficacy and extent of receptor desensitization, whereas the substituent electronegativity is correlated with ligand potency. NMR results show that the protein backbone amide chemical shift deviations correlate mainly with efficacy and extent of desensitization. Pronounced deviations occur at specific residues in the ligand-binding site and in the two helical segments that join the lobes by a disulfide bond. Experiments detecting conformational exchange show that micro- to millisecond timescale motions also occur near the disulfide bond and vary largely with efficacy and extent of desensitization. These results thus identify regions displaying structural and dynamical dissimilarity arising from differences in ligand-protein interactions and lobe closure that may play a critical role in receptor response. Furthermore, measures of line broadening and conformational exchange for a portion of the ligand-binding site correlate with ligand EC(50) data. These results do not have any correlate in the currently available crystal structures and thus provide a novel view of ligand-binding events that may be associated with agonist potency differences. PMID- 18387632 TI - Crystal structure of the polyextremophilic alpha-amylase AmyB from Halothermothrix orenii: details of a productive enzyme-substrate complex and an N domain with a role in binding raw starch. AB - The gene for a membrane-bound, halophilic, and thermostable alpha-amylase, AmyB, from Halothermothrix orenii was cloned and sequenced. The crystal structure shows that, in addition to the typical domain organization of family 13 glycoside hydrolases, AmyB carries an additional N-terminal domain (N domain) that forms a large groove--the N-C groove--some 30 A away from the active site. The structure of AmyB with the inhibitor acarbose at 1.35 A resolution shows that a nonasaccharide has been synthesized through successive transglycosylation reactions of acarbose. Unexpectedly, in a complex of wild-type AmyB with alpha cyclodextrin and maltoheptaose at 2.2 A resolution, a maltotetraose molecule is bound in subsites -1 to +3, spanning the cleavage point at -1/+1, with the -1 glucosyl residue present as a (2)S(o) skew boat. This wild-type AmyB complex was obtained in the presence of a large excess of substrate, a condition under which it is possible to capture Michaelis complexes, which may explain the observed binding across -1/+1 and ring distortion. We observe three methionine side chains that serve as "binding platforms" for glucosyl rings in AmyB, a seemingly rare occurrence in carbohydrate-binding proteins. The structures and results from the biochemical characterization of AmyB and AmyB lacking the N domain show that the N domain increases binding of the enzyme to raw starch. Furthermore, theoretical modeling suggests that the N-C groove can accommodate, spatially and chemically, large substrates such as A-starch. PMID- 18387633 TI - Invasion of tumorigenic HT1080 cells is impeded by blocking or downregulating the 37-kDa/67-kDa laminin receptor. AB - The 37-kDa/67-kDa laminin receptor precursor/laminin receptor (LRP/LR) acting as a receptor for prions and viruses is overexpressed in various cancer cell lines, and their metastatic potential correlates with LRP/LR levels. We analyzed the tumorigenic fibrosarcoma cell line HT1080 regarding 37-kDa/67-kDa LRP/LR levels and its invasive potential. Compared to the less invasive embryonic fibroblast cell line NIH3T3, the tumorigenic HT1080 cells display approximately 1.6-fold higher cell-surface levels of LRP/LR. We show that anti-LRP/LR tools interfere with the invasive potential of HT1080 cells. Anti-LRP/LR single-chain variable fragment antibody (scFv) iS18 generated by chain shuffling from parental scFv S18 and its full-length version immunoglobulin G1-iS18 reduced the invasive potential of HT1080 cells significantly by 37% and 38%, respectively. HT1080 cells transfected with lentiviral plasmids expressing small interfering RNAs directed against LRP mRNA showed reduced LRP levels by approximately 44%, concomitant with a significant decrease in the invasive potential by approximately 37%. The polysulfated glycans HM2602 and pentosan polysulfate (SP-54), both capable of blocking LRP/LR, reduced the invasive potential by 20% and 35%, respectively. Adhesion of HT1080 cells to laminin-1 was significantly impeded by scFv iS18 and immunoglobulin G1-iS18 by 60% and 68%, respectively, and by SP-54 and HM2602 by 80%, suggesting that the reduced invasive capacity achieved by these tools is due to the perturbation of the LRP/LR-laminin interaction on the cell surface. Our in vitro data suggest that reagents directed against LRP/LR or LRP mRNA such as antibodies, polysulfated glycans, or small interfering RNAs, previously shown to encompass an anti-prion activity by blocking or downregulating the prion receptor LRP/LR, might also be potential cancer therapeutics blocking metastasis by interfering with the LRP/LR-laminin interaction in neoplastic tissues. PMID- 18387634 TI - Crystallographic studies on multiple conformational states of active-site loops in pyrrolysyl-tRNA synthetase. AB - Pyrrolysine, a lysine derivative with a bulky pyrroline ring, is the "22nd" genetically encoded amino acid. In the present study, the carboxy-terminal catalytic fragment of Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS) was analyzed by X-ray crystallography and site-directed mutagenesis. The catalytic fragment ligated tRNA(Pyl) with pyrrolysine nearly as efficiently as the full length PylRS. We determined the crystal structures of the PylRS catalytic fragment in the substrate-free, ATP analogue (AMPPNP)-bound, and AMPPNP/pyrrolysine-bound forms, and compared them with the previously-reported PylRS structures. The ordering loop and the motif-2 loop undergo conformational changes from the "open" states to the "closed" states upon AMPPNP binding. On the other hand, the beta 7-beta 8 hairpin exhibits multiple conformational states, the open, intermediate (beta 7-open/beta 8-open and beta 7-closed/beta 8-open), and closed states, which are not induced upon substrate binding. The PylRS structures with a docked tRNA suggest that the active-site pocket can accommodate the CCA terminus of tRNA when the motif-2 loop is in the closed state and the beta 7-beta 8 hairpin is in the open or intermediate state. The entrance of the active-site pocket is nearly closed in the closed state of the beta 7-beta 8 hairpin, which may protect the pyrrolysyladenylate intermediate in the absence of tRNA(Pyl). Moreover, a structure-based mutational analysis revealed that hydrophobic residues in the amino acid-binding tunnel are important for accommodating the pyrrolysine side chain and that Asn346 is essential for anchoring the side-chain carbonyl and alpha-amino groups of pyrrolysine. In addition, a docking model of PylRS with tRNA was constructed based on the aspartyl-tRNA synthetase/tRNA structure, and was confirmed by a mutational analysis. PMID- 18387635 TI - Hyperglycemia and cognitive outcome after ischemic stroke. AB - BACKGROUND: Post-stroke hyperglycemia (HG) is associated with poor physical recovery, in particular in patients with cortical stroke. We tested whether HG is also associated with cognitive impairment after ischemic stroke. METHODS: We recruited patients from a prospective consecutive cohort with a first-ever supratentorial infarct. Neuropsychological examination included abstract reasoning, verbal memory, visual memory, visual perception and construction, language, and executive functioning. We related HG (glucose >7.0 mmol/L) to cognition and functional outcome (modified Barthel Index) at baseline and after 6 10 months, and to neurological deficit (National Institutes of Health Stroke Scale) and infarct size at baseline. In additional analyses cortical and subcortical infarcts were considered separately. RESULTS: Of 113 patients, 43 had HG (38%) and 55 had cortical infarcts (49%). Follow-up was obtained from 76 patients (68%). In the acute phase, in patients with cortical infarcts HG was associated with impaired executive function (B=-0.65; 95% confidence limits (CL): -1.3-0.00; p<0.05), larger lesion size (p<0.01), and more severe neurological deficits (p<0.01). These associations were not observed in patients with subcortical infarcts and the association between HG and cognitive functioning at follow-up was not significant in either group. CONCLUSIONS: In first-ever ischemic stroke, HG was not associated with impaired cognition after 6-10 months. In the acute phase of stroke HG was associated with impaired executive function, but only in patients with cortical infarcts. PMID- 18387636 TI - Contribution of slowly inactivating potassium current to delayed firing of action potentials in NG108-15 neuronal cells: experimental and theoretical studies. AB - The properties of slowly inactivating delayed-rectifier K+ current (I(K)(dr)) were investigated in NG108-15 neuronal cells differentiated with long-term exposure to dibutyryl cyclic AMP. Slowly inactivating I(K)(dr) could be elicited by prolonged depolarizations from -50 to +50 mV. These outward K+ currents were found to decay at potentials above -20 mV, and the decay became faster with greater depolarization. Cell exposure to aconitine resulted in the reduction of I(K)(dr) amplitude along with an accelerated decay of current inactivation. Under current-clamp recordings, a delay in the initiation of action potentials (APs) in response to prolonged current stimuli was observed in these cells. Application of aconitine shortened the AP initiation in combination with an increase in both width of spike discharge and firing frequency. The computer model, in which state dependent inactivation of I(K)(dr) was incorporated, was also implemented to predict the firing behavior present in NG108-15 cells. As the inactivation rate constant of I(K)(dr) was elevated, the firing frequency was progressively increased along with a shortening of the latency for AP appearance. Our theoretical work and the experimental results led us to propose a pivotal role of slowly inactivating I(K)(dr) in delayed firing of APs in NG108-15 cells. The results also suggest that aconitine modulation of I(K)(dr) gating is an important molecular mechanism through which it can contribute to neuronal firing. PMID- 18387637 TI - Chemical preconditioning effect of 3-nitropropionic acid in anesthetized rat heart. AB - Short ischemic episodes increase tolerance against subsequent severe ischemia in the heart. Nitropropionate (3-NP), an irreversible inhibitor of succinic dehydrogenase of the mitochondrial complex II, was shown to induce protective effect against ischemic brain injury. The aim of this study was to investigate the possible protective effect of 3-NP on regional ischemia in preconditioned rat heart in vivo. Hearts were assigned into three groups: first, in order to induce ischemic preconditioning (IP) 5 min ischemia separated by 10 min reperfusion protocol was used; second, non-preconditioned group was used as control; and third, 3-NP (20 mg/kg, i.p.) was injected 3 h before the surgical procedure in order to induce chemical preconditioning. In all these groups, 30 min regional ischemia was followed by 60 min reperfusion. Infarct size, bax expression, number of ventricular ectopic beats (VEB), duration of ventricular tachycardia (VT) and ventricular fibrillation (VF) were significantly decreased in ischemic preconditioning and 3-NP pretreatment groups, whereas bcl-2 values were not markedly changed in these groups during occlusion period. These results showed that in the anesthetized rat heart 3-NP induced chemical preconditioning by decreasing infarct size, number of VEB, duration of VT and VF. Protective effect is associated with via decreased production of bax protein expression. PMID- 18387638 TI - Neuroprotection achieved in the ischaemic rat cortex with L-kynurenine sulphate. AB - L-kynurenine is a metabolic precursor of kynurenic acid, which is one of the few known endogenous N-methyl-D-aspartate receptor inhibitors. In contrast with kynurenic acid, L-kynurenine is transported across the blood-brain barrier, and it may therefore come into consideration as a therapeutic agent in certain neurobiological disorders, e.g. ischaemia-induced events. The present study evaluated the effect of L-kynurenine administration (300 mg/kg i.p.) on the global ischaemic brain cortex both pre- and post-ischemic intervention. The statistical evaluation revealed that L-kynurenine administration beneficially decreased the number of neurones injured per mm(2) in the cortex, not only in the pre-treated animals, but also in those which received L-kynurenine after the ischaemic insult. It is concluded that even the post-traumatic administration of L-kynurenine may be of substantial therapeutic benefit in the treatment of global brain ischaemia. This is the first histological proof of the neuroprotective effect achieved by the post-traumatic administration of L-kynurenine in the global ischaemic cortex. PMID- 18387639 TI - A simple explanation for the low impact of border control as a countermeasure to the spread of an infectious disease. AB - A simple model for the effect of border control or travel restrictions is proposed. It can be used to predict the corresponding results in quite complex disease spread models and has the advantage of providing easy qualitative understanding of the effects of this kind of intervention. PMID- 18387640 TI - Keeping the goal in mind: prefrontal contributions to spatial navigation. PMID- 18387641 TI - Changing pattern of premature mortality burden over 6 years of rapid growth of the economy in suburban south-west China: 1998-2003. AB - BACKGROUND: This study was conducted in Kunming, the capital of Yunnan, a poor province in south-west China experiencing rapid economic growth. The study examined the short-term trend in premature mortality burden from common causes of death in a suburban region between 1998 and 2003. METHODS: Years of life lost (YLL) per 1000 population and mortality rate per 100,000 population were calculated from medical death certificates, and broken down by cause of death, sex and year without age weighting but with a discounting rate of 3%. RESULTS: Non-communicable diseases contributed over 80% of all causes of YLL, with a slightly increasing trend. The combined rate for communicable, maternal, prenatal and nutritional deficiencies declined from 4.7 to 2.4 per 1000 population. Remarkably, declining trends in YLL were also seen for chronic obstructive pulmonary disease, drug use and road traffic accidents, whereas increasing trends were seen for ischaemic heart disease (IHD) and liver cancer (males). The YLL rate for stroke, self-inflicted injuries, lung cancer and stomach cancer fluctuated over time. CONCLUSIONS: The region should focus on further control of IHD and liver cancer. PMID- 18387642 TI - Efficacy of toxic sugar baits against adult cistern-dwelling Anopheles claviger. AB - Sugar baits were used for mosquito control in cisterns at the building complex of Mar Saba Monastery in the Judean Hills, Israel. These cisterns provide fresh water for the monastery and are breeding and resting sites for Anopheles claviger (Meigen). The baits, including sucrose, fruit juice, dye marker and oral insecticide, were presented at the entrance of the cisterns and caused a drastic decline of A. claviger numbers in the neighbourhood. Pre-treatment catches of 16.8+/-4.0 to 33+/-8.1 females per trap decreased to between 2.5+/-1.3 and 0.3+/ 0.6 females per trap six nights after treatment. Numbers of males were initially 9.2+/-1.7 to 21.0+/-4.8 specimens per trap and decreased to between 1.0+/-0.4 and 0.5+/-0.2 per trap post treatment. In the control area, where baits without toxin were presented in cisterns, there were only small fluctuations in the mosquito population. Non-poisonous bait marked >95% of the mosquitoes exiting from a cistern in the control area. Following treatment, the number of human-landing mosquitoes decreased by more than ten-fold in the experimental area, whilst in the control area there was no significant decrease. PMID- 18387643 TI - Cervical cancer in Bangladesh: community perceptions of cervical cancer and cervical cancer screening. AB - We investigated the awareness of, and the attitude towards screening for, cervical cancer in Bangladesh. We performed a qualitative study using focus group discussions (FGD). The framework approach to qualitative analysis was used. The study was performed in the catchment areas of Addin Hospital, Jessore, Southern Bangladesh (peri-urban) and LAMB hospital, Parbatipur, North West Bangladesh (rural). A total of 220 men, women and adolescents participated in 28 FGDs. Awareness of cervical cancer was widespread. Knowledge about causes was often inadequate. The perceived consequences of cervical cancer were numerous and awareness of the need for cervical cancer treatment was present. Barriers to accessing care include: low priority for seeking help for symptoms, limited availability of health services and cost. Most women were unaware of the possibility of screening via speculum examination, which was considered acceptable to women (and men), as long as the examination was done by a female healthcare provider in an environment with sufficient privacy. In conclusion, adequate gynaecological services are not available or accessible for most women in rural and peri-urban Bangladesh. However, awareness of cervical cancer is widespread. Screening for cervical cancer in these communities is acceptable if done by a female healthcare provider under conditions with sufficient privacy. PMID- 18387644 TI - Synthesis of highly pure oxyphytosterols and (oxy)phytosterol esters Part I. Regioselective hydrogenation of stigmasterol: an easy access to oxyphytosterols. AB - The synthesis of several oxyphytosterols is described starting from stigmasterol, the key step being the regioselective hydrogenation of the 22-23 double bond of the latter. PMID- 18387645 TI - JWA is required for arsenic trioxide induced apoptosis in HeLa and MCF-7 cells via reactive oxygen species and mitochondria linked signal pathway. AB - Arsenic trioxide, emerging as a standard therapy for refractory acute promyelocytic leukemia, induces apoptosis in a variety of malignant cell lines. JWA, a novel retinoic acid-inducible gene, is known to be involved in apoptosis induced by various agents, for example, 12-O-tetradecanoylphorbol 13-acetate, N-4 hydroxy-phenyl-retinamide and arsenic trioxide. However, the molecular mechanisms underlying how JWA gene is functionally involved in apoptosis remain largely unknown. Herein, our studies demonstrated that treatment of arsenic trioxide produced apoptosis in HeLa and MCF-7 cells in a dose-dependent manner and paralleled with increased JWA expression. JWA expression was dependent upon generation of intracellular reactive oxygen species induced by arsenic trioxide. Knockdown of JWA attenuated arsenic trioxide induced apoptosis, and was accompanied by significantly reduced activity of caspase-9, enhanced Bad phosphorylation and inhibited MEK1/2, ERK1/2 and JNK phosphorylations. Arsenic trioxide induced loss of mitochondrial transmembrane potential was JWA-dependent. These findings suggest that JWA may serve as a pro-apoptotic molecule to mediate arsenic trioxide triggered apoptosis via a reactive oxygen species and mitochondria-associated signal pathway. PMID- 18387647 TI - Propofol pretreatment attenuates LPS-induced granulocyte-macrophage colony stimulating factor production in cultured hepatocytes by suppressing MAPK/ERK activity and NF-kappaB translocation. AB - Propofol (PPF), a widely used intravenous anesthetic for induction and maintenance of anesthesia during surgeries, was found to possess suppressive effect on host immunity. This study aimed at investigating whether PPF plays a modulatory role in the lipopolysaccharide (LPS)-induced inflammatory cytokine expression in a cell line of rat hepatocytes. Morphological observation and viability assay showed that PPF exhibits no cytotoxicity at concentrations up to 300 microM after 48 h incubation. Pretreatment with 100 microM PPF for 24 h prior to LPS stimulation was performed to investigate the modulatory effect on LPS induced inflammatory gene production. The results of semi-quantitative RT-PCR demonstrated that PPF pretreatment significantly suppressed the LPS-induced toll like receptor (TLR)-4, CD14, tumor necrosis factor (TNF)-alpha, and granulocyte macrophage colony-stimulating factor (GM-CSF) gene expression. Western blotting analysis showed that PPF pretreatment potentiated the LPS-induced TLR-4 downregulation. Flow cytometrical analysis revealed that PPF pretreatment showed no modulatory effect on the LPS-upregulated CD14 expression on hepatocytes. In addition, PPF pretreatment attenuated the phosphorylation of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and IkappaBalpha, as well as the nuclear translocation of NF-kappaB primed by LPS. Moreover, addition of PD98059, a MAPK kinase inhibitor, significantly suppressed the LPS induced NF-kappaB nuclear translocation and GM-CSF production, suggesting that the PPF-attenuated GM-CSF production in hepatocytes may be attributed to its suppressive effect on MAPK/ERK signaling pathway. In conclusion, PPF as an anesthetic may clinically benefit those patients who are vulnerable to sepsis by alleviating sepsis-related inflammatory response in livers. PMID- 18387646 TI - Chlordecone altered hepatic disposition of [14C]cholesterol and plasma cholesterol distribution but not SR-BI or ABCG8 proteins in livers of C57BL/6 mice. AB - Organochlorine (OC) insecticides continue to occur in tissues of humans and wildlife throughout the world although they were banned in the United States a few decades ago. Low doses of the OC insecticide chlordecone (CD) alter hepatic disposition of lipophilic xenobiotics and perturb lipid homeostasis in rainbow trout, mice and rats. CD pretreatment altered tissue and hepatic subcellular distribution of exogenous [(14)C]cholesterol (CH) equivalents 4 and 16 h after a bolus intraperitoneal (ip) injection of 5 ml corn oil/kg that contained 10 mg CH/kg. CD pretreatment altered tissue distribution of exogenously administered [(14)C]CH by decreased hepatic and renal accumulation, and increased biliary excretion up to 300%. Biliary excretion of polar [(14)C]CH metabolites was not altered by CD. CD pretreatment decreased subcellular distribution of [(14)C]CH equivalents in hepatic cytosol and microsomes and lipoprotein-rich fraction-to homogenate ratio. CD pretreatment increased the ratio of [(14)C]CH equivalents in high density lipoprotein (HDL) to that in plasma and reduced [(14)C]CH equivalents in the non-HDL fraction 4 h after a bolus lipid dose. CD pretreatment increased plasma non-HDL total CH by 80% 4 h after a bolus lipid dose. Scavenger receptor class B type I (SR-BI) and ATP-binding cassette transporter G8 (ABCG8) proteins were quantified by western blotting in hepatic membranes from control and CD treated mice. Liver membrane contents of SR-BI and ABCG8 proteins were unchanged by CD pretreatment. The data demonstrated that a single dose of CD altered CH homeostasis and lipoprotein metabolism. PMID- 18387648 TI - Inhibition of lung tumor colonization and cell migration with the disintegrin crotatroxin 2 isolated from the venom of Crotalus atrox. AB - Disintegrins are low molecular weight proteins (4-15 kDa) with an RGD binding region at their binding loop. Disintegrin and disintegrin-like proteins are found in the venom of four families of snakes: Atractaspididae, Elapidae, Viperidae, and Colubridae. This report describes the biological activity of a disintegrin, crotatroxin 2, isolated by a three-step chromatography procedure from the venom of the Western diamondback rattlesnake (Crotalus atrox). The intact molecular mass for crotatroxin 2 was 7.384 kDa and 71 amino acids. Crotatroxin 2 inhibited human whole blood platelet aggregation with an IC(50) of 17.5 nM, inhibited cell (66.3p) migration by 63%, and inhibited experimental lung tumor colonization in BALB/c mice at 1000 microg/kg. Our data suggest that while crotatroxin 2 inhibits platelet aggregation, cancer cell migration, and lung tumor colonization, it is done via different integrins. PMID- 18387649 TI - Analysis of filtration characteristics in submerged microfiltration for drinking water treatment. AB - Hollow fiber membranes have been widely employed for water and wastewater treatments. Nevertheless, understanding the filtration characteristics of hollow fiber membranes is complicated by the axial distributions of transmembrane pressure (TMP) and flux, which are key factors for both fouling control and module design. In this study, model equations to account for different fouling mechanisms were derived to analyze the performance of submerged hollow fiber systems with different conditions in terms of feed water characteristics and membrane material. A series of experiments with synthetic feed and raw water were carried out using hydrophilic and hydrophobic membrane modules. The model successfully fits the experimental results for synthetic feed as well as raw water. The major fouling mechanisms for filtration of raw water using hydrophilic and hydrophobic membranes are identified as cake formation and standard blocking, respectively. The model calculations indicate that the distributions of flux and cake (fouling) resistance are sensitive to the fiber length of the membrane. PMID- 18387650 TI - Uptake of polycyclic aromatic hydrocarbons by Trifolium pretense L. from water in the presence of a nonionic surfactant. AB - A greenhouse study examined plant uptake of phenanthrene and pyrene, as representatives of polycyclic aromatic hydrocarbons (PAHs), from an aqueous solution containing a nonionic surfactant Tween 80. The uptake was conducted with 1.0 mg l(-1) phenanthrene and 0.12 mg l(-1) pyrene under a wide range of Tween 80 concentrations (0-105.6 mg l(-1)). Tween 80 at the test concentrations did not show any apparent phytotoxity toward the growth of red clover (Trifolium pretense L.). At concentrations generally lower than 13.2 mg l(-1), Tween 80 enhanced the plant uptake based on the concentrations and PCFs (plant concentration factors) of these two PAHs. When present at higher concentrations, Tween 80 inhibited the uptake of both PAH compounds by the tested plant. The maximal plant uptake was observed at 6.6 mg l(-1) Tween 80, in which PAH concentrations and PCFs were 18 115% higher than those in Tween 80-free controls. The total mass removal (off take) of phenanthrene and pyrene by root or shoot increased initially and decreased thereafter with the increase in Tween 80 concentrations. Although shoot biomass was evidently larger than root, the off-take was much higher in root than shoot because of the larger root concentrations of these chemicals. Results from this study show promises for the potential efficacy of enhanced phytoremediation in PAH contaminated sites using surfactant amendment. PMID- 18387651 TI - Hominin-carnivore interactions during the Chinese Early Paleolithic: taphonomic perspectives from Xujiayao. AB - The ability of archaic Homo sapiens to survive in more northerly latitudes was contingent on securing a regular source of animal fat and protein. We present a taphonomic study that examines how successful these hominins were at acquiring these food sources during the latter part of the Early Paleolithic in Northeast Asia. This study focuses on the long bone midshaft surface modifications observed on the faunal remains from Xujiayao, a middle-late Pleistocene open-air site located at 40 degrees latitude in the western Nihewan Basin, northern China. The faunal assemblage is dominated by equid remains. Analysis of the percussion, tooth, and cut mark frequencies on the long bone midshafts demonstrates that the Xujiayao hominins had primary access to high utility (meat-bearing and marrow rich) long bones. Investigation of the dual-patterned (tooth-marked and butchery marked) bone fragments suggests that hominins were under little pressure from competing carnivores to abandon their kills. The lack of significant differences between the size of fragments with only percussion-marks and those with only tooth-marks supports these findings. Fragmentation ratios indicate that forelimbs were more intensively processed than hind limbs. Based on the water rounding and abrasion data, the Xujiayao assemblage is likely of autochthonous origin. Since the age of Xujiayao is still in question, we can only conclude that archaic Homo sapiens were successful predators of large game in Northeast Asia some time during the latter part of the Early Paleolithic. PMID- 18387652 TI - Coevolution of telomerase activity and body mass in mammals: from mice to beavers. AB - Telomerase is repressed in the majority of human somatic tissues. As a result human somatic cells undergo replicative senescence, which plays an important role in suppressing tumorigenesis, and at the same time contributes to the process of aging. Repression of somatic telomerase activity is not a universal phenomenon among mammals. Mice, for example, express telomerase in somatic tissues, and mouse cells are immortal when cultured at physiological oxygen concentration. What is the status of telomerase in other animals, beyond human and laboratory mouse, and why do some species evolve repression of telomerase activity while others do not? Here we discuss the data on telomere biology in various mammalian species, and a recent study of telomerase activity in a large collection of wild rodent species, which showed that telomerase activity coevolves with body mass, but not lifespan. Large rodents repress telomerase activity, while small rodents maintain high levels of telomerase activity in somatic cells. We discuss a model that large body mass presents an increased cancer risk, which drives the evolution of telomerase suppression and replicative senescence. PMID- 18387653 TI - Exposure of mother-child and postpartum woman-infant pairs to DDT and its metabolites in Tianjin, China. AB - 1,1,1-trichloro-2,2'-bis(4-chlorophenyl)ethane (DDT) exhibits its long persistence in the environment, unusual bioaccumulation, effects on wildlife, and the possibility of long-term adverse effects on human health, especially reproductive toxicity. Despite the prohibition of most persistent organochlorine pesticides in China, the presence of organochlorine residue, including DDT, has been widely indicated in environmental substance. However, scarce information is available about accumulative levels of DDT in human tissues in China. To evaluate levels of DDT and its potential effects on women and children's health in a Chinese pesticide-exposed area, we recruited 50 pairs of mother-child and 50 postpartum women, and determined the levels of total DDT and its four main metabolites (p,p'-DDE, p,p'-DDT, p,p'-DDD, o,p'-DDT) in venous blood, breast milk and umbilical blood cord by gas chromatography. Accordingly, data on reproductive outcomes of mothers and postpartum women and healthy status of children and infants were gathered through a questionnaire and medical examinations. Furthermore, we also assayed the DDT levels of some environmental samples (soil, food, milk, et al.). The levels of DDT in children's blood were higher than that in the women's. As compared to breast milk, the umbilical blood cord and the ventral fat individually demonstrated a significantly lower and higher level of DDT in the postpartum women. DDT was lower in milk and crucian carp than in the soil near the chemical plant. p,p'-DDT and p,p'-DDE were the main metabolites of DDT. Our findings suggested the cumulative effect of DDT in human body in Tianjin, China. PMID- 18387654 TI - Superficial venous thrombosis: prevalence of common genetic risk factors and their role on spreading to deep veins. AB - INTRODUCTION: Superficial venous thrombosis (SVT) has been considered for a long time a limited clinical condition with a low importance, but this approach has changed in recent years, when several studies demonstrated spreading to deep veins occurring from 7.3 to 44%, with high prevalence of pulmonary embolism. MATERIALS AND METHODS: To evaluate the prevalence of genetic risk factors for VTE in patients suffering from SVT on both normal and varicose vein, and to understand their role on spreading to deep veins, we studied 107 patients with SVT, without other risk factors. Ultrasound examination was performed, and the presence of FV Leiden, Prothrombin G20210A mutation, and MTHFR C677T mutation was researched. RESULTS: In the patients where SVT occurred in normal veins, the presence of FV Leiden was 26.3% of the non-spreading and 60% of the spreading to deep veins SVT; Prothrombin mutation was found in 7.9% of the former case and in 20% of the latter; MTHFR C677T mutation was found respectively in 23.7% and 40%. In the patients with SVT on varicose veins, the presence of these factors was less evident (6.7%, 4.4% and 6.7% respectively), but their prevalence was considerably higher (35.7%, 7.4% and 21.4% respectively) in SVT spreading to deep veins than in non-spreading. CONCLUSIONS: Our data demonstrate the high prevalence of these mutations, especially FV Leiden and associations, in patients with SVT on normal veins and their role in the progression to deep vein system. PMID- 18387655 TI - Improved fibrinolytic activity during exercise may be an effect of the adipocyte derived hormones leptin and adiponectin. AB - INTRODUCTION: Physical activity is associated with improved fibrinolytic activity and reduced risk for cardiovascular disease. High levels of leptin and low levels of adiponectin, both adipocyte-derived hormones, or adipokines, are related to dysfibrinolysis and risk for cardiovascular disease. In this study, we explored if improved fibrinolytic activity during exercise could be linked to changes in leptin and adiponectin levels. MATERIALS AND METHODS: Twenty healthy men (mean age 36 years) participated in a 14-day long skiing expedition in the Swedish mountains. They were randomly assigned to either a 40% or a 30% fat-based diet. Anthropometry, lipids, fibrinolytic activity (PAI-1 activity, tPA activity and mass) and adipokines (leptin and adiponectin) were measured before, during and six weeks after the expedition. RESULTS: PAI-1 activity and circulating levels of leptin decreased whereas levels of adiponectin increased during exercise. The fall in PAI-1 activity showed a strong linear association with changes in leptin and adiponectin levels (p = 0.001 and p < 0.001, respectively). Changes in leptin and adiponectin levels were independent of decreasing waist circumference. However, the association between anthropometric measures and adipokines changed considerably during the expedition. Adiponectin was weakly and negatively associated with BMI at baseline. In contrast, there was a strong positive association between adiponectin and BMI after two weeks of exercise, whereas the association between leptin and BMI became less pronounced. In addition, increasing leptin and decreasing adiponectin levels were associated with increasing PAI-1 activity during the six weeks following the expedition. After six weeks of normal activity, fibrinolytic activity and hormone levels returned towards baseline levels. CONCLUSION: Heavy exercise induced improved fibrinolytic activity, which was associated independently with changes in circulating levels of the adipocyte-derived hormones leptin and adiponectin. Improved fibrinolytic activity (and reduced risk for cardiovascular disease) related to physical activity could possibly be mediated by leptin and adiponectin. PMID- 18387656 TI - The long-term prognostic value of multiple biomarkers following a myocardial infarction. AB - INTRODUCTION: We assessed the long-term prognostic value of multiple cardiac biomarkers after an acute myocardial infarction (MI) in order to evaluate a multimarker approach to risk stratification. MATERIAL AND METHODS: Blood samples from 298 patients hospitalized with a myocardial infarction were subsequently tested for NT-proBNP, hsCRP, MMP-9, PAPP-A, MPO, sCD40L and FM. RESULTS: During the median follow-up period of 45 months, 83 patients suffered at least one TnT- positive event. In the unadjusted analysis NT-proBNP predicted future ACS or cardiac death with a hazard ratio (HR) of 1.83 (95% confidence interval (CI), 1.17-2.87, p=0.009) in Q4 as compared to the three lower quartiles (Q1-3). However, NT-proBNP was dependent on chronic heart failure and HDL-cholesterol in the stepwise multivariable model, with a hazard ratio (HR) in Q4 of 1.38 (95% CI, 0.82-2.33, p=0.229). The other biomarkers were not found to be related to the primary event following the index MI. CONCLUSION: In a patient population consisting of 298 subjects hospitalized with a MI, a multimarker approach with NT proBNP, hsCRP, MMP-9, PAPP-A, MPO, sCD40L and FM rendered no additional prognostic information beyond conventionally used stratification tools in the acute phase. However, this does not preclude clinical valuable prognostic information by a biomarker such as NT-proBNP. PMID- 18387657 TI - Platelet-stimulating effects of oxidized LDL are not attributable to toxic properties of the lipoproteins. AB - One prominent feature of oxidized LDL (OxLDL) is their ability to activate human platelets and effects of OxLDL on platelet function have been shown to depend on the chemical mechanisms that form the basis for the oxidation process. In this regard, the possibility that the observed platelet-stimulating properties of OxLDL might be a direct consequence of cytotoxic effects mediated by these lipoproteins merits further investigation, as experimental strategies to overcome the toxic effects of OxLDL towards a variety of different cell types did not yield conclusive results. In the present work, we show that copper-oxidized LDL mediate severe toxic effects towards a macrophage cell line (decrease in both the number of adherent cells and the amount of incorporated tritiated thymidine, induction of apoptosis and subsequent loss of membrane integrity)--effects that are presumably attributable to products emerging from lipid peroxidation. When added to resting human platelets, copper oxidized LDL stimulate platelets but are not able to trigger an aggregation response on their own. In contrast, hypochlorite-oxidized LDL are able to trigger platelet aggregation, but do not mediate toxic effects towards nucleated cells. Even in the absence of exogenous antioxidants, these lipoproteins mediate cytostatic effects but do not negatively affect cell viability. As a conclusion, platelet-activating effects of oxidatively modified LDL are not attributable to toxic properties of the lipoproteins and this finding might expand possibilities for therapeutical intervention. PMID- 18387658 TI - Impact of body mass index on biochemical recurrence rates after radical prostatectomy: an analysis utilizing propensity score matching. AB - OBJECTIVES: To investigate the significance of body mass index (BMI) as an independent predictor of biochemical recurrence in men treated with surgery for clinically localized adenocarcinoma of the prostate. METHODS: A total of 1877 obese patients who underwent radical prostatectomy were matched to overweight and normal-weight patients in a 1:1 ratio on the basis of propensity scores. This resulted in an overall study population of 5631 men. Clinicopathologic characteristics and biochemical recurrence outcomes after surgery were compared between the three BMI cohorts. RESULTS: Normal-weight patients exhibited lower grade disease compared with overweight and obese patients (P = 0.021). Lower BMI was also significantly associated with lower rates of positive surgical margins (P <0.001) and extraprostatic extension (P <0.001). Body mass index was not associated with lymph node involvement (P = 0.226) or seminal vesicle invasion (P = 0.142). Body mass index, age, biopsy Gleason score, preoperative prostate specific antigen level, and clinical tumor stage were independent predictors of biochemical recurrence (P <0.001). CONCLUSIONS: Propensity score-based matched analyses indicate that higher BMI is associated with adverse pathologic findings and is a strong independent predictor of biochemical recurrence after radical prostatectomy. These results support the hypothesis that inherent differences may exist in the biological properties of prostate cancer in obese men compared with normal-weight men. Therefore, BMI is an important criterion to consider during subsequent decision making and counseling of patients with prostate cancer. PMID- 18387659 TI - Correlation between duration of varicocele and apoptosis in testicular tissue in an experimental model. AB - OBJECTIVES: To experimentally demonstrate the negative effect of a waiting period for treatment in patients with varicocele. METHODS: A varicocele group of 30 rats and a sham group of 10 were used. All rats in the varicocele groups underwent ligation in the left testicle proximal to the site at which the internal spermatic vein drains into the renal vein to constrict the vein by 50%. In the sham group, the renal vein was dissected but not ligated. One month later, the first 15 rats in the experimental group and the first 5 rats in the sham group underwent laparotomy and bilateral orchiectomy. At 2 months postoperatively, the same procedures were performed on the second group of rats (15 varicocele rats and 5 sham rats). The apoptotic index was evaluated by group and interval after varicocele or sham operation. For apoptosis detection, modification of in situ end labeling was used. RESULTS: At 1 and 2 months after creation of the varicocele or sham operation, the apoptosis rate in the left varicocele group was significantly greater than in the sham group (P <0.001) or in the right testicles (P <0.001). Although an increase in the apoptosis rate was seen in the second month compared with the first month, the difference was not statistically significant (P = 0.875). The histopathologic finding of atrophy was more significant in the second month than in the first month in the varicocele group. CONCLUSIONS: The results of our study have indicated that the increase in apoptosis at the end of the first month in the varicocele group was significant and that this apoptosis continued with time. Because the histopathologic injury, which is parallel to apoptosis, increased, we believe that treatment in the early phase of varicocele is important for prevention of probable injury. PMID- 18387660 TI - Expression of chicken ovalbumin upstream promoter-transcription factor I (COUP TFI) in bladder transitional cell carcinoma. AB - OBJECTIVES: An analysis of differentially expressed genes (DEGs) between bladder transitional cell carcinoma (TCC) and the surrounding urothelium to help identify what lies behind the mechanism of multifocal tumor development has not yet been performed. We sought to find a new DEG related to the development of bladder TCC. METHODS: Thirty-nine bladder TCC tissues paired with normal-appearing urothelium tissues obtained from the same patient were used as subjects. Initially, we compared the messenger RNA (mRNA) profiles between normal-appearing urothelium and TCC tissue of 1 patient by using annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR) and selective amplification of family members (SAFM) PCR to identify potential DEGs. To validate the results of the ACP data, reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on those of all 39 patients. RESULTS: Among the several DEGs discovered in the ACP data, 1 DEG was chosen as the candidate for the RT-PCR, that is present or markedly upregulated in normal-appearing urothelial tissue compared with TCC tissue. Gene sequence searching revealed that this DEG is chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI). Downregulation of COUP-TFI mRNA expression in TCC tissue compared to normal-appearing urothelium tissue of the same patient, irrespective of tumor stage and grade, was confirmed by RT-PCR in 39 patients. CONCLUSIONS: Our results suggest that the loss of COUP-TFI may play a role in the transition from normal epithelium to TCC. Further characterization of the COUP-TFI gene is expected to give us informations about bladder TCC tumorigenesis. PMID- 18387663 TI - Vesicovaginal fistula formation in patients with Stage IVA cervical carcinoma. PMID- 18387661 TI - Effect of DNA methylation on identification of aggressive prostate cancer. AB - OBJECTIVES: Biochemical (prostate-specific antigen) recurrence of prostate cancer after radical prostatectomy remains a major problem. Better biomarkers are needed to identify high-risk patients. DNA methylation of promoter regions leads to gene silencing in many cancers. In this study, we assessed the effect of DNA methylation on the identification of recurrent prostate cancer. METHODS: We studied the methylation status of 15 pre-specified genes using methylation specific polymerase chain reaction on tissue samples from 151 patients with localized prostate cancer and at least 5 years of follow-up after prostatectomy. RESULTS: On multivariate logistic regression analysis, a high Gleason score and involvement of the capsule, lymph nodes, seminal vesicles, or surgical margin were associated with an increased risk of biochemical recurrence. Methylation of CDH13 by itself (odds ratio 5.50, 95% confidence interval [CI] 1.34 to 22.67; P = 0.02) or combined with methylation of ASC (odds ratio 5.64, 95% CI 1.47 to 21.7; P = 0.01) was also associated with an increased risk of biochemical recurrence. The presence of methylation of ASC and/or CDH13 yielded a sensitivity of 72.3% (95% CI 57% to 84.4%) and negative predictive value of 79% (95% CI 66.8% to 88.3%), similar to the weighted risk of recurrence (determined from the lymph node status, seminal vesicle status, surgical margin status, and postoperative Gleason score), a powerful clinicopathologic prognostic score. However, 34% (95% CI 21% to 49%) of the patients with recurrence were identified by the methylation profile of ASC and CDH13 rather than the weighted risk of recurrence. CONCLUSIONS: The results of our study have shown that methylation of CDH13 alone or combined with methylation of ASC is independently associated with an increased risk of biochemical recurrence after radical prostatectomy even considering the weighted risk of recurrence score. These findings should be validated in an independent, larger cohort of patients with prostate cancer who have undergone radical prostatectomy. PMID- 18387664 TI - P16 expression in relation to human papillomavirus in liquid-based cervical smears. AB - OBJECTIVE: At present, a simple and reliable cervical cancer screening test remains to be perfected. As overexpression of the protein p16 is correlated with the presence of high-risk HPV in malignant cervical lesions, this protein has been proposed as a surrogate marker of high-risk HPV infection in cervical cancer screening. Since high-risk viral DNA integration is necessary for neoplastic progression, we aimed to examine the expression of p16 in relation to the physical status of HPV (integrated or episomal) on liquid-based cervical smears. METHODS: For each of the 241 liquid-based cervical smear included in our study, we realized a Pap test. Residual cells were processed for in situ hybridization with mucosal HPV DNA probes and for immunocytochemistry with an anti-p16 antibody. Integrated or episomal copies of HPV DNA were detected as dotted or diffuse signals, respectively. RESULTS: In high-grade intraepithelial lesions, both the integrated form of high-risk HPV and overexpression of p16 were detected. However, we observed the presence of some p16-positive/HPV-negative normal and ASCUS smears. Moreover, some p16-negative ASCUS smears and low-grade intraepithelial lesions harbored episomal high-risk HPV. CONCLUSION: If p16 was used as a surrogate marker of high-risk HPV infection, some women would be scored negative in spite of the presence of high-risk HPV. These women are more likely to undergo cancer progression, but no follow-up would be proposed to them in that screening pathway. A possible compromise for the triage of abnormal cervical smears should be a combination of both HPV and p16 testing. PMID- 18387665 TI - Early embryo development in the elephant assessed by serial ultrasound examinations. AB - The elephant has an extraordinary long pregnancy, lasting 21 months. However, knowledge on embryo development is limited. To date, only single morphological observations of elephant embryo development associated with placentation are available, all lacking correlation to gestational age. The present study describes morphological characteristics of early embryo development in the elephant with exact biometric staging. Six pregnancies in five Asian and one African elephants with known conception dates were followed by 2D and 3D ultrasound, covering the embryonic period from ovulation to day 116 post ovulation. The embryonic vesicle was earliest observed was on day 50 p.o. The proper embryo was not detected until day 62 p.o. Embryonic heartbeat was first observed on day 71 p.o. The allantois, which became visible as a single sacculation on day 71 p.o. was subdivided in four compartments on day 76 p.o. By day 95 p.o., head, rump, front and hind legs were clearly distinguished. Between days 95 and 103 p.o. the choriovitelline placenta was replaced by the chorioallantoic placenta. A physiological midgut herniation was transiently present between days 95 and 116 p.o. On the basis of the late appearance of the embryonic vesicle, delayed implantation in the elephant is discussed. The study provides a coherent description of elephant embryonic development, formation of the extraembryonic organs and their role in placenta formation, all of which are of interest for both comparative evolutionary studies and the improvement of assisted reproduction techniques. PMID- 18387666 TI - Risk of myocardial infarction and nucleoside analogues. PMID- 18387668 TI - Abacavir and the potential risk of myocardial infarction. PMID- 18387669 TI - Placental anti-oxidant gene polymorphisms, enzyme activity, and oxidative stress in preeclampsia. AB - The etiology and pathophysiology of preeclampsia are not fully understood. However, oxidative stress has been strongly linked to the occurrence of this multi-system disease. This has led to many theories of the pathogenesis of preeclampsia involving placental oxidative stress. In this study, we hypothesized that polymorphisms of anti-oxidant genes in the placental tissue contributed to susceptibility to preeclampsia. Polymorphisms in copper/zinc superoxide dismutase (CuZn-SOD), manganese superoxide dismutase (MnSOD), glutathione-S-transferase M1 (GSTM1), and glutathione-S-transferase T1 (GSTT1) in the umbilical cord tissue were assayed by polymerase chain reaction (PCR) in 23 nulliparous preeclampsia cases and 32 nulliparous normotensive controls. Corresponding enzyme activity levels and an oxidative stress biomarker (8-isoprostane) of the placental tissue were also measured. In addition, maternal plasma 8-isoprostane levels were also determined. Our results showed that no significant differences in polymorphism frequency of the tested genes, enzyme activity levels or 8-isoprostane levels in the placental tissue were detected between the cases and controls. However, maternal plasma 8-isoprostane level was significantly higher in the cases than in the controls (105.8 vs. 27.9 pg/ml, p=0.03). In conclusion, our study showed that polymorphisms of CuZn-SOD, MnSOD, GSTM1 and GSTT1 in the placental tissue were not associated with preeclampsia. PMID- 18387670 TI - Altered uterine perfusion is involved in fetal outcome of diabetic rats. AB - Maternal diabetes affects the development of the offspring by altering the uterine environment. We aimed to investigate the extent to which the blood flow (measured as Tissue Perfusion Units; TPU) to implantation sites and the expression of developmentally important genes in the offspring are affected by maternal diabetes. We measured mRNA levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), Bcl-2 associated X protein (Bax), B cell lymphoma protein (Bcl-2), tumor suppressor protein-53 (p53), paired box protein-3 (Pax-3) and vascular endothelial growth factor-A (Vegf-A). Moreover, we studied the effect on uterine blood flow (TPU) and the expression of the genes exerted by embryonic maldevelopment (malformation or resorption). Streptozotocin induced diabetic (D) and non-diabetic (N) pregnant rats were used in the study. Blood flow (TPU) to implantation sites was measured by a laser Doppler flow meter, and gene expression was analyzed by RT-PCR. Maternal diabetes caused increased blood flow (TPU) to implantation sites compared with normal pregnancy. Furthermore, implantation sites of D rats containing malformed offspring showed impaired growth and decreased blood flow (TPU) compared with their littermates at all gestational days. Resorbed offspring from both N and D rats displayed increased blood flow (TPU) compared with their non-resorbed littermates. Moreover, we found that maternal diabetes causes decreased expression of genes involved in the oxidative stress defense system (CuZnSOD in non-malformed D11 embryos, MnSOD at all gestational time points, ECSOD and Gpx-1 at GD11-GD15, CAT and Gpx-2 at GD15), decreased expression of Pax-3 at GD11, and increased expression of Vegf-A at all gestational time points. We conclude that both maternal metabolism and embryonic developmental state affect the blood flow (TPU) to the implantation site. Maternal diabetes causes decreased expression of anti oxidative enzymes and enhanced angiogenesis in the offspring in rats. PMID- 18387671 TI - Cord compression may rapidly influence the expression of placental angiogenic genes in pre-eclampsia. AB - Gene expression studies have demonstrated the altered expression level of placental angiogenesis related genes in severe pre-eclampsia (PE). In cord compression, the transportation of oxygen from the placenta to the fetus is blocked, and it is speculated that during blockade the originally hypoxic placenta may become hyperoxic. We compared the placental gene expression profiles of one pre-eclamptic patient with cord compression (the index patient) to the profiles of patients with PE and those of normal pregnancy controls (including one woman with cord compression). The gene expression of the cord compression PE patient resembled that observed in the normal pregnancies. We hypothesize that umbilical blockade may in a short period of time lead to placental hyperoxia, which in turn has an effect on angiogenic gene expression profile. PMID- 18387672 TI - Neurotoxicity of MPTP to migrating neuroblasts: studies in acute and subacute mouse models of Parkinson's disease. AB - The acute or subacute administration of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) has been widely used in C57BL/6 mice to develop models of Parkinson's disease (PD). The loss of dopaminergic neurons is suggested to be mediated by a mechanism of nonapoptotic cell death or by apoptosis. In recent years, the notion that the neurotoxicity of MPTP is restricted to dopaminergic neurons in the substantia nigra (SN) has been challenged. Here, we provide evidence of rapid cell death in the subventricular zone (SVZ) and rostral migratory stream (RMS) in the adult C57BL/6 mouse brain in response to acute or subacute treatment with MPTP. Significant terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) of fragmented DNA was observed at 24 h (or 1 day) after the last injection in the acute model or after the first injection in the subacute model. Ultrastructural analysis confirmed that dying cells displayed an apoptotic morphology. Using a double labeling method, we demonstrated that the phenotype of the cells undergoing apoptosis is that of migrating neuroblasts. This is further supported by evidence of a subsequent loss of migrating neuroblasts. The results raise the possibility that migrating neuroblasts in the SVZ and RMS may be more vulnerable to MPTP than nigrostriatal dopaminergic neurons in the SN, and the death of migrating neuroblasts may be a primary event in the mouse model of PD. Furthermore, our data suggests that the death and subsequent loss of migrating neuroblasts in the acute or subacute model probably lead to a decreased potential for neurogenesis to some extent. PMID- 18387673 TI - Expressed emotion, perceived criticism and 10-year outcome of depression. AB - Outcome studies of patients suffering from depression indicate a high relapse rate and a tendency towards chronicity. A family atmosphere characterised by expressed emotion (EE) and perceived criticism (PC) was reported to be a robust predictor of outcome in affective disorders. The aim of the study was to analyze the association between EE, PC and long-term outcome of depression. Fifty inpatients with major depression were followed up 1, 2 and 10 years after discharge from the hospital. After 10 years, 26 patients (56.5%) had at least one recurrence. Spousal EE, assessed with the Five-Minute Speech Sample (FMSS), and the Perceived Criticism Index (PCI) were not significant prognostic factors for depression outcome. The significance of EE and PC for the long-term course of depression has to be questioned. The findings indicate a need to examine factors that may partly mediate the effect of EE and PC on the outcome of depression. PMID- 18387674 TI - Gastric Helicobacter species as a cause of feline gastric lymphoma: a viable hypothesis. AB - Gastric Helicobacter spp. are associated with chronic inflammation and neoplastic transformation in humans as well as domestic and laboratory species. The present study examined the association of Helicobacter heilmannii (Hhe) infection in pet cats with feline gastric mucosa associated lymphoid tissue (MALT) lymphoma. Tissues were collected via gastric biopsy or at necropsy from 47 pet cats with clinical signs of gastrointestinal disease, including vomiting and inappetance, and classified as gastritis (14/47), lymphoma (31/37), or normal (2/47). Tissues positive for argyrophilic organisms with Warthin-Starry stain (29/47) were assessed by fluorescent in situ hybridization (FISH) for the presence of Hhe strains 1-4 as well as with a fifth probe that detected Helicobacter salomonis, Helicobacter bizzozeronii, or Helicobacter felis. A significant association of positive Warthin-Starry status with Hhe infection was found in cases of sick cats (22/29; p<0.05 by Chi-square; chi(2)=7.034). Interestingly, a significant association between Hhe status and a diagnosis of lymphoblastic or lymphocytic lymphoma was observed as well in a subset of 24 Warthin-Starry positive lymphoma cases: of lymphoblastic lymphoma cases, 13/17 were positive for Hhe (p<0.05; chi(2)=4.854). Hhe strains 2 and 4 were most commonly found (18/29 and 17/29, respectively) among sick cats, although a higher than expected number of cats was also positive for Hhe1, which initial reports have described as rare in cats and common in humans. The association found between a positive Hhe status with the presence of feline gastric lymphoma, especially lymphoblastic lymphoma, argues for the need to conduct prospective studies to better identify the frequency and strain distribution of Hhe infection in both healthy and clinically ill cats, particularly those cats with gastric lymphoma. PMID- 18387675 TI - Development and in vitro characterization of canine CD40-Ig. AB - We recently reported that blockade of the CD40-CD154 ligand interaction with the cross-reacting mouse anti-human CD154 antibody, 5c8, together with donor-specific transfusion led to enhanced but not completely successful engraftment in a canine model of DLA-identical marrow transplantation after 100cGy total body irradiation (TBI). In order to improve the transplantation outcomes, we sought to develop a canine-specific reagent. To that end, we fused the extracellular domain of the canine CD40 with a mouse IgG2a Fc tail and tested the immunosuppressive effectiveness of the fusion protein in mixed leukocyte reactions. The extracellular domain of canine CD40 was fused with the Fc portion of mouse IgG2a in a pcDNA3.1+vector. Dhfr-deficient CHO cells were co-transfected with the CD40 Ig vector and a dhfr-containing vector. Stable, high producing clones were selected under increasing methotrexate concentrations. The fusion protein was purified, tested in mixed leukocyte reactions, and its immunosuppressive effect compared to that of the anti-CD154 antibody 5c8. The transfected cell line produced a CD40-Ig dimer whose identity was confirmed by mass spectroscopy. The purified canine CD40-Ig blocked mixed leukocyte reactions at a concentration of 1nM, which was 10 times more effective than the anti-CD154 antibody. Canine CD40 Ig is more immunosuppressive than the anti-human CD154 antibody 5c8 in canine mixed leukocyte reactions and may be more effective in vivo in a model of marrow transplantation. PMID- 18387676 TI - Juvenile nasopharyngeal angiofibroma stages I and II: a comparative study of surgical approaches. AB - OBJECTIVE: The aim of this study was to compare different surgical options used for removal of stages I and II juvenile nasopharyngeal angiofibromas (JNAs). Treatment morbidity was evaluated through blood loss, surgery duration, postoperative hospitalization and outcome. Moreover, an effort was made to explore the role and limits of endoscopic surgery. MATERIALS AND METHODS: 20 male patients (mean age 14.9 years) were treated for JNA using three different surgical approaches in the Department of Otolaryngology of the University of Athens between May 1998 and January 2007. 9 patients were managed using endoscopic approach, while 5 were treated through midfacial degloving. A transpalatal approach was performed in remaining 6 patients. Preoperative angiography with embolization was performed in all 9 patients who underwent endoscopic removal and in 3 patients treated by midfacial degloving technique. RESULTS: Findings demonstrated that endoscopic approach, assisted by preoperative embolization, lead to less intraoperative blood loss, shorter duration of surgical procedure, shorter length of hospital stay and no complications, compared with the conventional techniques. CONCLUSIONS: Our data suggest that with proper patient selection, endoscopic resection of stages I and II JNA, when it is performed after embolization of the feeding vessels, is remarkably bloodless and precise and may be preferable to traditional open approaches. PMID- 18387677 TI - Management of infant ranula. AB - OBJECTIVE: Many surgical techniques to manage ranulas have been described in the literature. Ranula of infant patients was rare. Few studies have described the approach toward management in infant patients. METHODS: Eleven infant patients were treated for intraoral ranulas. The methods of treatment included aspiration of mucus, marsupialization and excision of the ranula and the ipsilateral sublingual gland. All cases were performed aspiration of mucus and observed for 6 months; and the marsupialization were recommended if the ranula recurred; the surgical resection of ipsilateral sublingual gland were performed if the ranula recurred when infant patients was about 1-year-old. These patients were followed up at least 24 months. RESULTS: Age of presentation ranged from 2 days to 3 months. There were six females (54.55%) and five males (45.45%). All cases presented simple (introral) ranula. Excision ranula with sublingual gland was performed on seven patients (63.64%) while marsupialization was performed on two patients (18.18%) and two patients (18.18%) were aspiration of the mucus of ranula and no recurrence. There were no recurrent lesions in all cases. CONCLUSION: Conservative treatment of infant ranula maybe includes observation for 6 months for spontaneous resolution. The methods for observation is performed the aspiration of mucus and marsupialization. The resection of ipsilateral sublingual gland is recommended if ranula recurred for infant patients about 1 year-old. We believe that it is safe that the submandibular duct and complete sublingual nerve are dissected before the sublingual gland is removed. PMID- 18387678 TI - Viral nucleic acid stabilization by RNA extraction reagent. AB - In the collection of field materials to test for the presence of arboviruses, samples must be appropriately maintained to detect arboviral nucleic acids. In austere field conditions this is often difficult to achieve because, during routine specimen processing, storage, and shipping viral RNA degradation could result in detection failure. RNA extraction reagents, while used commonly for their intended purpose of stabilizing RNA during the extraction process, have not been assessed fully for their potential to stabilize RNA before extraction. The potential for virus stabilization at varying temperatures and periods of time remains unknown. Accordingly, the ability of buffer AVL (Qiagen, Valencia, CA), an RNA extraction reagent, to stabilize viral suspensions of dengue, Venezuelan equine encephalitis and Rift Valley fever viruses was evaluated. The ability of buffer AVL to stabilize each viral suspension was examined at 32, 20, 4, and -20 degrees C. RNA in samples placed in buffer AVL was stable for at least 48h at 32 degrees C and refrigerating samples prolonged stabilization. Additionally, placing the sample/buffer AVL mixture at either 4 or -20 degrees C stabilized samples for at least 35 days. When combined with the ability of buffer AVL to inactivate viral samples, this provides the ability to collect and handle potentially infectious samples in a safe way that also provides sample stabilization. PMID- 18387679 TI - A rapid, high-throughput vaccinia virus neutralization assay for testing smallpox vaccine efficacy based on detection of green fluorescent protein. AB - Virus neutralization remains a vital tool in assessment of vaccine efficacy for smallpox in the absence of animal smallpox models. In this regard, development of a rapid, sensitive, and high-throughput vaccinia neutralization assay has been sought for evaluating alternative smallpox vaccines, use in bridging studies, as well as understanding the effects of anti-viral immunotherapeutic regimes. The most frequently used method of measuring vaccinia virus neutralization by plaque reduction is time, labor, and material intensive, and therefore limiting in its utility for large scale, high-throughput analysis. Recent advances provide alternative methods that are less labor intensive and higher throughput but with limitations in reagents needed and ease of use. An innovative neutralization assay is described based on a modified Western Reserve vaccinia vector expressing green fluorescent protein (WR-GFP) and an adherent cell monolayer in multi-well plate format. The assay is quick, accurate, provides a large dynamic range and is well suited for large-scale vaccination studies using standard adherent cell lines. PMID- 18387680 TI - Responses of chronically contaminated biofilms to short pulses of diuron. An experimental study simulating flooding events in a small river. AB - An experimental study was undertaken to highlight the potential ecotoxicological impact of the herbicide diuron on biofilms during flooding events in a small river (Morcille) in the Beaujolais vineyard area (France). We investigated the responses of chronically contaminated biofilms exposed to short-term pulses (3 h) of diuron. Biofilms were grown in indoor microcosms that were either non contaminated or exposed to low-level chronic contamination, and not exposed, or exposed to single or double pulses of two environmental concentrations (7 and 14 microg L(-1)) of diuron. Exposure to pollution and its impact on biofilms were assessed by measuring pesticide concentrations in biofilms, biomass parameters (chl a, AFDW), community structure (using 18S and 16S rDNA gene analysis by DGGE, and HPLC pigment analysis to target eukaryotes, bacteria and photoautotrophs, respectively) and by performing a physiological test. Control biofilms displayed very low diuron concentrations, whereas the herbicide was found in the contaminated biofilms. Nevertheless, diuron concentrations were not higher in the pulsed biofilms than in the non-pulsed ones. AFDW and chl ain vivo fluorescence increased in both microcosms during the experiment and biomass was higher in chronically exposed biofilms than in control ones. The impact on biomass was higher for the control double-pulsed biofilms than for the non-pulsed ones. Carbon incorporation by the chronically exposed biofilms was greater during the first 28 days of growth than during the first 28 days of growth in the control biofilms. Both single and double pulses inhibited carbon incorporation of all biofilm communities, especially of the control ones. Short-term inhibition of photosynthesis was never significantly different in exposed and non-exposed biofilms. Few differences in the pigment structure were found between chronically exposed and control biofilms, but pulses impacted on the pigment structure of all biofilm communities. Bacterial structural differences were observed between single-pulsed and non-pulsed biofilms, but not between double-pulsed and non pulsed biofilms. The different pulses affected the eukaryotic community structures of the control biofilms, but not of the chronically exposed ones. Unlike the bacterial communities, the control eukaryotic communities were structurally different from the chronically exposed ones. This preliminary experimental study indicates that exposure to environmental concentrations of diuron and other agricultural contaminants and further exposure to diuron can have measurable effects on small river biofilm communities. The effects of a pulsed acute exposure to diuron on biofilms depended on whether the biofilms had previously been exposed to the same stressors or not. PMID- 18387681 TI - Lymphangiogenesis promotes inflammation and neointimal hyperplasia after adventitia removal in the rat carotid artery. AB - Lymphatic vessels exist in adventitia in the atherosclerotic coronary artery and play an important role in the inflammatory and immune response. After adventitia removal, the carotid wall of rat model showed significantly increased ratio of intimal to medial area (I/M ratio), the number of adventitial lymphatic vessels (Ad-LV) and microvessels (Ad-MV), and macrophage index and expression of VEGF-C, VEGFR-3, PDGF-B and PDGFR-beta. The I/M ratio was significantly correlated with Ad-LV and macrophage index but not Ad-MV. These results suggest that adventitial lymphangiogenesis is stimulated by growth factors released by inflammatory cells in vasculature after adventitia removal, and these neogenetic lymph vessels in turn promote intimal inflammation and hyperplasia, probably via delivery and activation of inflammatory cells. PMID- 18387682 TI - Maternal employment, breastfeeding, and health: evidence from maternity leave mandates. AB - Public health agencies around the world have renewed efforts to increase the incidence and duration of breastfeeding. Maternity leave mandates present an economic policy that could help achieve these goals. We study their efficacy, focusing on a significant increase in maternity leave mandates in Canada. We find very large increases in mothers' time away from work post-birth and in the attainment of critical breastfeeding duration thresholds. We also look for impacts of the reform on self-reported indicators of maternal and child health captured in our data. For most indicators we find no effect. PMID- 18387683 TI - A split-protein sensor for studying protein-protein interaction in mycobacteria. AB - The study of protein function in living cells is an essential complement to genomics, yet method development does not always keep pace with sequencing. Experimental techniques for the genus mycobacteria are relatively underdeveloped, though seventeen genomes have been sequenced. "Split-Trp" is a split-protein sensor used to detect protein-protein interactions in tryptophan auxotrophic Saccharomyces cerevisiae, but the principles behind the sensor should allow it to function in a broad range of microbial hosts. Here we introduce Split-Trp to Escherichia coli and Mycobacterium smegmatis and demonstrate that this system is a simple assay for protein interaction in both organisms. PMID- 18387667 TI - Use of nucleoside reverse transcriptase inhibitors and risk of myocardial infarction in HIV-infected patients enrolled in the D:A:D study: a multi-cohort collaboration. AB - BACKGROUND: Whether nucleoside reverse transcriptase inhibitors increase the risk of myocardial infarction in HIV-infected individuals is unclear. Our aim was to explore whether exposure to such drugs was associated with an excess risk of myocardial infarction in a large, prospective observational cohort of HIV infected patients. METHODS: We used Poisson regression models to quantify the relation between cumulative, recent (currently or within the preceding 6 months), and past use of zidovudine, didanosine, stavudine, lamivudine, and abacavir and development of myocardial infarction in 33 347 patients enrolled in the D:A:D study. We adjusted for cardiovascular risk factors that are unlikely to be affected by antiretroviral therapy, cohort, calendar year, and use of other antiretrovirals. FINDINGS: Over 157,912 person-years, 517 patients had a myocardial infarction. We found no associations between the rate of myocardial infarction and cumulative or recent use of zidovudine, stavudine, or lamivudine. By contrast, recent-but not cumulative-use of abacavir or didanosine was associated with an increased rate of myocardial infarction (compared with those with no recent use of the drugs, relative rate 1.90, 95% CI 1.47-2.45 [p=0.0001] with abacavir and 1.49, 1.14-1.95 [p=0.003] with didanosine); rates were not significantly increased in those who stopped these drugs more than 6 months previously compared with those who had never received these drugs. After adjustment for predicted 10-year risk of coronary heart disease, recent use of both didanosine and abacavir remained associated with increased rates of myocardial infarction (1.49, 1.14-1.95 [p=0.004] with didanosine; 1.89, 1.47-2.45 [p=0.0001] with abacavir). INTERPRETATION: There exists an increased risk of myocardial infarction in patients exposed to abacavir and didanosine within the preceding 6 months. The excess risk does not seem to be explained by underlying established cardiovascular risk factors and was not present beyond 6 months after drug cessation. PMID- 18387684 TI - Site-directed, Ligase-Independent Mutagenesis (SLIM) for highly efficient mutagenesis of plasmids greater than 8kb. AB - Modifying the Site-directed, Ligase-Independent Mutagenesis (SLIM) protocol from a single reaction mode to a two-reaction mode enables highly efficient mutagenesis of plasmid constructs that exceed 8kb. This modified approach reduces the complexity of the PCR step and is optimised for generation of heteroduplexes from long PCR products. The two-reaction mode SLIM has 92% efficiency. PMID- 18387685 TI - Multi-method approach indicates no presence of sub-lethally injured Listeria monocytogenes cells after mild heat treatment. AB - Application of mild inactivation treatments follows an increasing trend in the food industry and is often combined with sub-optimal intrinsic product conditions to ensure appropriate level of microbial safety. Listeria monocytogenes was subjected to mild heat treatment (20 min at 60 degrees C) and subsequently exposed to various mild preservation conditions based on increased NaCl concentration and decreased pH. Recovery and resuscitation of L. monocytogenes cells were studied using various methods. Using 12-fold Most Probable Number (MPN) method no difference in the amount of recovered cells under adverse conditions was noted between heat-treated and non-treated L. monocytogenes cells. Time-to-detection method using on-line OD measurements showed that heat-treated L. monocytogenes cells reached detection limit faster in acidified media and NaCl supplemented media in comparison with non-heated control cells. Flow cytometry (FCM) analysis using 5-6-carboxyfluorescein diacetate (cFDA) and propidium iodide (PI) staining showed presence of low numbers of viable cells. Overall, there was no indication of sub-lethal injury in L. monocytogenes cells after mild heat treatment. PMID- 18387686 TI - Comparative examination and validation of ELISA test systems for Salmonella typhimurium diagnosis of slaughtering pigs. AB - The most frequently isolated Salmonella serotype from pork in Germany is S. typhimurium, especially phagetype DT 104. The monitoring programs on Salmonella in swine are based on enzyme-linked immunoadsorbent assay (ELISA) detecting antibodies in serum or meat juice. These serological results are used to classify swine herds in three categories to assess the hygienic status of farm regarding Salmonella infection in pigs. The object of this study was the comparative evaluation of four indirect Salmonella ELISA tests approved in Germany to detect Salmonella typhimurium infection of swine. Three tests (A-C) are based on LPS antigen and directed against specific IgG-antibodies. The fourth test (D) bases on a whole-cell-lysate antigen and discriminates between Salmonella specific IgA , IgM- and IgG-antibodies. In a longitudinal study sixteen 6 weeks old weaning pigs were orally infected with S. typhimurium DT 104. During an observation period of 138d clinical and bacteriological parameters were monitored and serum samples obtained at regular intervals as well as meat juice samples taken at slaughter were examined by the respective ELISA systems. Study results reveal that all tested ELISA systems are able to detect S. typhimurium infection in pigs in both sample matrices, blood serum and meat juice whereas test D showed the highest sensitivity to detect Salmonella antibodies in pigs. The sensitivity to detect Salmonella antibodies varied between tests A and C according to the used cut-off (test specific cut-off vs. recommended surveillance cut-off) resulting in a change of seroprevalence and hence may influence the Salmonella status of the farm. PMID- 18387687 TI - Cloning and subcellular localization of the phosphoprotein and nucleocapsid proteins of Potato yellow dwarf virus, type species of the genus Nucleorhabdovirus. AB - We have cloned and characterized mRNAs corresponding to the phosphoprotein (P) and nucleocapsid (N) genes of the sanguinolenta strain of Potato yellow dwarf virus (PYDV). The P and N messenger RNAs both begin with a common AAACA pentanucleotide and are 1546nt and 962nt in length, and capable of encoding 52kDa and 31kDa proteins, respectively. The N mRNA contains a 12nt 5' non-translated sequence (NTS) and a 83nt 3'-NTS. Similarly, the P mRNA has a 19nt 5'-NTS and a 125nt 3'-NTS. Primary structure analyses revealed three potential phosphorylation sites in the P protein and six in the N protein. Despite a lack of predictable nuclear localization signals (NLSs) in either protein, transient expression of the P and N proteins in N. benthamiana showed that both proteins are targeted exclusively to nuclei. Phylogenetic analyses showed that PYDV is most closely related to Maize mosaic virus and Taro vein chlorosis virus, which also lack predictable NLSs in their N proteins. The present data further distinguish PYDV from SYNV and suggest that, together, these viruses serve to provide a more comprehensive view of rhabdovirus cell biology, which can be studied in a common host plant. PMID- 18387688 TI - Liver regeneration and recanalization time course following reversible portal vein embolization. AB - BACKGROUND/AIMS: Permanent portal vein embolization (PVE) is a widely practised technique. The use of an absorbable material would be safer in clinical situations in which the embolized liver is not resected. We evaluated the efficiency of reversible PVE in terms of liver regeneration and analyzed the precise time course of portal recanalization. METHODS: Nine monkeys underwent PVE of the left and right anterior portal branches using powdered absorbable material. Repeated portograms were carried out until complete revascularization of the embolized liver. Hepatocyte proliferation rates were assessed by BrdU incorporation. Liver segment volumes were determined by CT scans performed before embolization, then 1 month and 1 year after embolization. RESULTS: Reversible PVE induced significant hepatocyte proliferation in the non-embolized segments (13.5+/-1.0%, 10.5+/-0.8% and 9.1+/-2.0% of cells on days 3, 5 and 7, respectively). One month after the embolization, the non-embolized liver volume had increased from 38.4+/-1.3% to 54.8+/-0.5% of total liver volume. Proximal and complete revascularization occurred 6-8 and 12-16 days, respectively. CONCLUSIONS: Reversible PVE efficiently induces liver regeneration. The use of absorbable material avoids long-term liver scarring. Such material may be suitable for several clinical indications, including cell transplantation. PMID- 18387689 TI - Comparing methods for full versus single technology appraisal: a case study of docetaxel and paclitaxel for early breast cancer. AB - OBJECTIVE: The aim of this study is to compare and contrast the reports of the rapid National Institute for Health and Clinical Excellence (NICE) single technology (STA) process with the full Health Technology Assessment Report (TAR) for the same technologies (docetaxel and paclitaxel for the adjuvant treatment of early breast cancer). METHODS: Documentary analysis was used to compare three reports. A data extraction form was developed to record information on the decision problem addressed in each report, literature searching methods, the clinical effectiveness review, cost-effectiveness analysis, conclusions and issues of concern. RESULTS: The decision problems and search strategies for the STAs differed from those reported in the TAR. Not all trials of clinical effectiveness included in the TAR were reported by manufacturers. The methods and assumptions used to assess the cost-effectiveness of the technologies were generally similar between reports, although only one common health economic comparison was presented across the three reports. The TAR supports the STA results for docetaxel but suggested that the manufacturer's cost-effectiveness estimates for paclitaxel were highly optimistic. CONCLUSIONS: Although the STA process provides the opportunity for rapid appraisal of new technologies, there remains uncertainty concerning the extent to which STAs adequately address the specific decision problem under consideration. PMID- 18387690 TI - The impact of the introduction of market incentives on occupational health services and occupational health professionals: experiences from The Netherlands. AB - OBJECTIVES: This paper describes the reform of the regulations on safety and health in the Netherlands towards a more competitive market and its impact on occupational health services (OHSs) and the health professionals over the period 1994-2005. Aims are to identify the crucial factors that bring about the intended effects (such as lower disability rates) and to evaluate the outcomes from the perspective of the occupational health professional. The paper contributes to the discussion of how the professionals could define and contain their professional identity and credibility in competitive circumstances. METHODS: Open interviews were completed with 12 key persons and secondary analyses were made on documents and various monitors. RESULTS: The reform changed the OHS safety market fundamentally. OHSs were transformed from medium sized regional units into business organizations mostly operating on a national level. Private insurance companies became key players. Only after the development of an effective social infrastructure, however, intended effects (lower absenteeism and disability) occurred. Occupational health professionals were initially opposed but by redefining their professional domain and identity, they finally succeeded in gaining negotiating power in order to preserve and develop expertise and professional integrity. CONCLUSIONS: The effectiveness of the introduction of market incentives depends strongly on their social embeddedness. Health professionals should adapt their strategy to the conditions of the competitive market, in order to preserve a credible and professional identity. PMID- 18387691 TI - Inflammatory cytokines increase extracellular procathepsin D in permanent and primary endothelial cell cultures. AB - The protease cathepsin D (Cath D) and its proteolytically inactive proform, procathepsin D (ProCath D), turned out to be multifunctional within and outside the cell. Elevated levels of ProCath D occur in malignant tumors and in organs under chronic inflammation. One important source for this increase of ProCath D might be endothelial cells. Here we examined the expression of Cath D in the human endothelial cell line EA.hy 926 and in primary endothelial cells isolated from human umbilical cord veins (HUVEC). After serum-free incubation with or without human interferon-gamma (hIFN-gamma) and/or human tumor necrosis factor alpha (hTNF-alpha) immature and mature Cath D forms were examined in cell extracts and in cell-conditioned medium concentrates by Western blotting. Lysates of EA.hy 926 cells as well as of HUVEC contained active Cath D as two-chain form, but only negligible amounts of ProCath D and Cath D intermediates. Yet both endothelial cell cultures accumulated ProCath D in their conditioned media in the absence of any stimulus. The treatment with hIFN-gamma and/or hTNF-alpha had little effect on intracellular levels of Cath D, whereas the cytokine stimulation increased the extracellular presence of ProCath D in both endothelial cell cultures. The extracellular increase of ProCath D was not related to induction of apoptosis, as validated by cleaved caspase-3 in cell lysates. Acidification of cytokine-treated media converted ProCath D into Cath D, which was associated with cathepsin-like activity using a fluorogenic substrate-linked assay. We conclude, in vitro, endothelial cells are a cytokine-dependent source for extracellular ProCath D. PMID- 18387692 TI - A volunteer feeding assistance program can improve dietary intakes of elderly patients--a pilot study. AB - Malnutrition is prevalent in elderly hospitalized patients and has been associated with longer lengths of stay (LOS), higher rates of complications and increased hospital costs. Feeding assistance has traditionally been the role of nurses, however with an ageing population and an ever-increasing workload there may not be sufficient time to ensure the nutritional care of all patients. A program in which trained volunteers assist, socialize and feed nutritionally vulnerable patients at lunch on weekdays has been initiated in a major suburban hospital in Sydney. The pilot study reported here aimed to evaluate the lunchtime assistance program in terms of dietary intakes by comparing data from weekdays (with volunteers) and that from weekends (no volunteers). Nine patients (mean age+/-S.D.: 89+/-4.6 years) participated in the study. Observations and weighed plate waste were recorded for each patient at lunch on two weekdays and the following two weekend days. When volunteers were present, the average protein intake increased by 10.1g at lunch (p<0.05) and 10.7 g over the whole day (p<0.05). There was also a trend to increased energy intake. Observations indicated that the volunteers, when compared to the nurses, socialized more with patients, encouraged them to eat more often and spent more time feeding them. Trialing volunteer assistance in a larger study would be useful. PMID- 18387693 TI - Healthcare workers' knowledge of the carriage of meticillin-resistant Staphylococcus aureus by patients: the first step in controlling spread. PMID- 18387694 TI - Preventing infections associated with the insertion of central venous catheters: are we getting it right? PMID- 18387696 TI - Opinion on Millar et al., 'Are meticillin-resistant Staphylococcus aureus bloodstream infection targets fair to those with other types of healthcare associated infection or cost-effective?'. PMID- 18387695 TI - Impact of preoperative screening for meticillin-resistant Staphylococcus aureus by real-time polymerase chain reaction in patients undergoing cardiac surgery. AB - We report a significant reduction in the number of surgical site infections (SSIs) due to meticillin-resistant Staphylococcus aureus (MRSA) in patients undergoing cardiac surgery after the introduction of preoperative screening using a same-day polymerase chain reaction (PCR) test. This was an observational cohort study set in a cardiac surgery unit based in southwest England. We studied 1462 patients admitted for cardiac surgery between October 2004 and September 2006. The IDI MRSA PCR test was used preoperatively to screen 765 patients between October 2005 and September 2006. Patients identified as carriers were treated with nasal mupirocin ointment and topical triclosan for five days, with single dose teicoplanin instead of flucloxacillin as perioperative antibiotic prophylaxis. The rate of SSI following cardiac surgery in this group was compared to 697 patients who underwent surgery without screening between October 2004 and September 2005. After introduction of PCR screening, the overall rate of SSI fell from 3.30% to 2.22% with a significant reduction in the rate of MRSA infections (relative risk reduction: 0.77; 95% confidence interval: 0.056-0.95). PCR screening combined with suppression of MRSA at the time of cardiac surgery is feasible in routine clinical practice and is associated with a significant reduction in subsequent MRSA SSIs. PMID- 18387697 TI - Community-associated meticillin-resistant Staphylococcus aureus in injecting drug users and the homeless in south London. PMID- 18387698 TI - Changes in the cost and management of emergency department on-call coverage: evidence from a longitudinal statewide survey. AB - STUDY OBJECTIVE: We measure changes in the prevalence and magnitude of stipends and other payments for taking emergency call during a 2-year period for hospitals in Oregon and evaluate the ways in which hospitals are limiting services and assessing policy options. METHODS: This was a longitudinal, standardized, e-mail based survey of chief executive officers from all hospitals with emergency departments (EDs) in Oregon (N=56). The first wave was conducted in the summer of 2005; a follow-up survey was conducted in summer 2006. Hospitals reported on-call payments made to 8 selected specialties. RESULTS: Among 56 Oregon hospitals with EDs, 43 responded to our survey in both 2005 and 2006, representing a 77% response rate. Among 54 specialties receiving stipends in 2006, the average stipend was $18,324. Total annual stipend payments increased by 84%, from an average of $227,000 per hospital in 2005 to $487,000 per hospital in 2006. In Oregon, between 2004 and 2006, 67% of hospitals lost the ability to provide coverage for at least 1 specialty on a 24-hour, 7-day-a-week basis. Approximately half of hospitals (49%) manage this lack of coverage by transferring patients to other hospitals on a case-by-case, ad hoc basis. CONCLUSION: The cost of maintaining on-call coverage is increasing in Oregon, raising concerns about hospital financing and a degradation of the emergency services. There has not been a systematic response to on-call shortages, with patient transfers primarily managed in an ad hoc, case-by-case basis. PMID- 18387699 TI - Forecasting emergency department crowding: a discrete event simulation. AB - STUDY OBJECTIVE: To develop a discrete event simulation of emergency department (ED) patient flow for the purpose of forecasting near-future operating conditions and to validate the forecasts with several measures of ED crowding. METHODS: We developed a discrete event simulation of patient flow with evidence from the literature. Development was purely theoretical, whereas validation involved patient data from an academic ED. The model inputs and outputs, respectively, are 6-variable descriptions of every present and future patient in the ED. We validated the model by using a sliding-window design, ensuring separation of fitting and validation data in time series. We sampled consecutive 10-minute observations during 2006 (n=52,560). The outcome measures--all forecast 2, 4, 6, and 8 hours into the future from each observation--were the waiting count, waiting time, occupancy level, length of stay, boarding count, boarding time, and ambulance diversion. Forecasting performance was assessed with Pearson's correlation, residual summary statistics, and area under the receiver operating characteristic curve. RESULTS: The correlations between crowding forecasts and actual outcomes started high and decreased gradually up to 8 hours into the future (lowest Pearson's r for waiting count=0.56; waiting time=0.49; occupancy level=0.78; length of stay=0.86; boarding count=0.79; boarding time=0.80). The residual means were unbiased for all outcomes except the boarding time. The discriminatory power for ambulance diversion remained consistently high up to 8 hours into the future (lowest area under the receiver operating characteristic curve=0.86). CONCLUSION: By modeling patient flow, rather than operational summary variables, our simulation forecasts several measures of near-future ED crowding, with various degrees of good performance. PMID- 18387700 TI - Out-of-hospital continuous positive airway pressure ventilation versus usual care in acute respiratory failure: a randomized controlled trial. AB - STUDY OBJECTIVE: Continuous positive airway pressure ventilation (CPAP) in appropriately selected patients with acute respiratory failure has been shown to reduce the need for tracheal intubation in hospital. Despite several case series, the effectiveness of out-of-hospital CPAP has not been rigorously studied. We performed a prospective, randomized, nonblinded, controlled trial to determine whether patients in severe respiratory distress treated with CPAP in the out-of hospital setting have lower overall tracheal intubation rates than those treated with usual care. METHODS: Out-of-hospital patients in severe respiratory distress, with failing respiratory efforts, were eligible for the study. The study was approved under exception to informed consent guidelines. Patients were randomized to receive either usual care, including conventional medications plus oxygen by facemask, bag-valve-mask ventilation, or tracheal intubation, or conventional medications plus out-of-hospital CPAP. The primary outcome was need for tracheal intubation during the out-of-hospital/hospital episode of care. Mortality and length of stay were secondary outcomes of interest. RESULTS: In total, 71 patients were enrolled into the study, with 1 patient in each group lost to follow-up after refusing full consent. There were no important differences in baseline physiologic parameters, out-of-hospital scene times, or emergency department diagnosis between groups. In the usual care group, 17 of 34 (50%) patients were intubated versus 7 of 35 (20%) in the CPAP group (unadjusted odds ratio [OR] 0.25; 95% confidence interval [CI] 0.09 to 0.73; adjusted OR 0.16; 95% CI 0.04 to 0.7; number needed to treat 3; 95% CI 2 to 12). Mortality was 12 of 34 (35.3%) in the usual care versus 5 of 35 (14.3%) in the CPAP group (unadjusted OR 0.3; 95% CI 0.09 to 0.99). CONCLUSION: Paramedics can be trained to use CPAP for patients in severe respiratory failure. There was an absolute reduction in tracheal intubation rate of 30% and an absolute reduction in mortality of 21% in appropriately selected out-of-hospital patients who received CPAP instead of usual care. Larger, multicenter studies are recommended to confirm this observed benefit seen in this relatively small trial. PMID- 18387701 TI - Advanced modeling strategies in emergency medicine research: is the pendulum swinging? PMID- 18387702 TI - Recurrence of primary headache disorders after emergency department discharge: frequency and predictors of poor pain and functional outcomes. AB - STUDY OBJECTIVE: We determine the frequency of moderate or severe headache during the first 24 hours after an emergency department (ED) visit for a primary headache disorder (such as migraine or tension-type headache), determine the burden of headache during the 3 months after the ED visit, and identify predictors of poor pain and functional outcomes after ED discharge for each of these periods. METHODS: In this prospective cohort study, we enrolled headache patients during their initial ED visit, interviewed them by using a standardized questionnaire, and followed them by telephone 24 hours and 3 months after ED discharge. Two emergency physicians classified all headaches according to criteria established by the International Headache Society, using a valid questionnaire and a reproducible technique. RESULTS: During an 18-month period, we enrolled 309 primary headache disorder patients in the cohort. The most common primary headache diagnoses assigned to patients were migraine, tension-type headache, and unclassifiable recurrent headache disorder. We successfully obtained follow-up in 94% of patients 24 hours after ED discharge and in 94% 3 months after ED discharge. Moderate or severe headache was present within 24 hours of ED discharge in 31% (95% confidence interval [CI] 25% to 38%) of migraine patients, 19% (95% CI 9% to 36%) of tension-type headache patients, and 27% (95% CI 18% to 38%) of the unclassifiable headache patients. Multiple functionally impairing headaches occurred during the 3 months after ED discharge in 37% of migraine patients (95% CI 30% to 44%), 38% of tension-type headache patients (95% CI 23% to 54%), and 26% of the unclassifiable headache patients (95% CI 17% to 37). After multivariate adjustment, independent predictors of poor 24-hour outcomes were severe baseline pain, baseline nausea, screening positive for depression, and longer duration of headache; the independent predictor of poor 3-month outcomes was Medicaid insurance. CONCLUSION: Regardless of type of primary headache disorder, ED headache patients frequently experience pain and functional impairment during the hours and months after discharge. PMID- 18387703 TI - Risk stratification of community-acquired pneumonia: what does all of this mean? PMID- 18387704 TI - Delayed defibrillation caused by unexpected ECG artifact. AB - Incorrect lead selection and unexpected ECG artifact during an attempted resuscitation after inhospital cardiac arrest resulted in undetected lack of cardiac monitoring for approximately 13 minutes. The patient was finally countershocked and regained a spontaneous pulse but was determined to have experienced profound neurologic damage and died shortly after being extubated. This type of failure may be common, particularly with older monitor/defibrillators. Caregivers, health care organizations, and device manufacturers should be aware of this potential problem and institute preventive measures. PMID- 18387705 TI - Thinking the unthinkable. PMID- 18387706 TI - Evidence-based emergency medicine/critically appraised topic. Is fluid therapy associated with cerebral edema in children with diabetic ketoacidosis? AB - STUDY OBJECTIVE: Diabetic ketoacidosis is the most common cause of morbidity and mortality in children with type I diabetes mellitus, and cerebral edema is the leading cause of pediatric diabetic ketoacidosis death. Excessive intravenous fluid administration has been implicated as a cause of cerebral edema. We perform an evidence-based emergency medicine review assessing the association of intravenous fluid hydration and cerebral edema. METHODS: We searched MEDLINE and EMBASE for comparative studies. Because of the low incidence of cerebral edema, we included observational studies in our review. We sought studies including patients younger than 18 years and with diabetic ketoacidosis. We defined cerebral edema, using clinical, radiographic, pathologic criteria, or treatment for intracranial pressure. RESULTS: Three studies met criteria for this review, of which 2 used fluid volume/body weight per unit of time, and the third measured cumulative total volume during hourly periods. The first 2 studies showed no statistically significant association between intravenous fluid administration and cerebral edema. The third study showed a significant association (odds ratio 6.55; 95% confidence interval 1.38 to 30.97) between the total volume (uncorrected for body weight) of infused fluid and the risk of cerebral edema. CONCLUSION: Accepting the inherent limitations of observational studies, we found a lack of strong or consistent results implicating rate or volume of fluid administration as a precipitant cause of cerebral edema in patients with diabetic ketoacidosis. PMID- 18387707 TI - Medical response to a radiologic/nuclear event: integrated plan from the Office of the Assistant Secretary for Preparedness and Response, Department of Health and Human Services. AB - The end of the Cold War led to a reduced concern for a major nuclear event. However, the current threats from terrorism make a radiologic (dispersal or use of radioactive material) or nuclear (improvised nuclear device) event a possibility. The specter and enormousness of the catastrophe resulting from a state-sponsored nuclear attack and a sense of nihilism about the effectiveness of a response were such that there had been limited civilian medical response planning. Although the consequences of a radiologic dispersal device are substantial, and the detonation of a modest-sized (10 kiloton) improvised nuclear device is catastrophic, it is both possible and imperative that a medical response be planned. To meet this need, the Office of the Assistant Secretary for Preparedness and Response in the Department of Health and Human Services, in collaboration within government and with nongovernment partners, has developed a scientifically based comprehensive planning framework and Web-based "just-in time" medical response information called Radiation Event Medical Management (available at http://www.remm.nlm.gov). The response plan includes (1) underpinnings from basic radiation biology, (2) tailored medical responses, (3) delivery of medical countermeasures for postevent mitigation and treatment, (4) referral to expert centers for acute treatment, and (5) long-term follow-up. Although continuing to evolve and increase in scope and capacity, current response planning is sufficiently mature that planners and responders should be aware of the basic premises, tools, and resources available. An effective response will require coordination, communication, and cooperation at an unprecedented level. The logic behind and components of this response are presented to allow for active collaboration among emergency planners and responders and federal, state, local, and tribal governments. PMID- 18387708 TI - Ovariectomy increases neuronal amyloid-beta binding alcohol dehydrogenase level in the mouse hippocampus. AB - Ovarian hormone decline after menopause may influence cognitive performance and increase the risk for Alzheimer's disease (AD) in women. Amyloid-beta peptide (Abeta) has been proposed to be the primary cause of AD. In this study, we examined whether ovariectomy (OVX) could affect the levels of cofactors Abeta binding alcohol dehydrogenase (ABAD) and receptor for advanced glycation endproducts (RAGE), which have been reported to potentiate Abeta-mediated neuronal perturbation, in mouse hippocampus, correlating with estrogen and Abeta levels. Female ICR mice were randomly divided into ovariectomized or sham operated groups, and biochemical analyses were carried out at 5 weeks after the operation. OVX for 5 weeks significantly decreased hippocampal 17beta-estradiol level, while it tended to reduce the hormone level in serum, compared with the sham-operated control. In contrast, OVX did not affect hippocampal Abeta(1-40) level, although it significantly increased serum Abeta(1-40) level. Furthermore, we demonstrated that OVX increased hippocampal ABAD level in neurons, but not astrocytes, while it did not affect RAGE level. These findings suggest that the expression of neuronal ABAD depends on estrogen level in the hippocampus and the increase in serum Abeta and hippocampal ABAD induced by ovarian hormone decline may be associated with pre-stage of memory deficit in postmenopausal women and Abeta-mediated AD pathology. PMID- 18387709 TI - Whole genome analysis in a consanguineous family with early onset Alzheimer's disease. AB - Early-onset Alzheimer's disease (EOAD) is a clinically and genetically heterogeneous condition in which the typical features appear significantly earlier in life (before 65 years). Mutations in three genes (PSEN1, PSEN2, and APP) have been identified in autosomal dominant forms of EOAD. However, in about 50% of Mendelian cases and in most of the sporadic EOAD patients, no mutations have been found. We present clinical characteristics of an Israeli family comprising two affected siblings with EOAD born to neurologically healthy parents who were first cousins (both parents died after 90 years old). Sequence analysis of PSEN1, PSEN2, APP, TAU, PGRN, and PRNP failed to reveal any mutations in the affected siblings. Because the disease in this family is consistent with an autosomal recessive mode of inheritance we identified all homozygous regions identical by descent (IBD) in both siblings, by high-density SNP genotyping. We provide here the first catalog of autozygosity in EOAD and suggest that the regions identified are excellent candidate loci for a recessive genetic lesion causing this disease. PMID- 18387710 TI - Synthesis and antimicrobial activities of a new series of 4-S-[4(1)-amino-5(1) oxo-6(1)-substituted benzyl-4(1),5(1)-dihydro-1(1),2(1),4(1)-triazin-3 yl]mercaptoacetyl-3-arylsydnones. AB - The synthesis of some 4-S-(4(1)-amino-5(1)-oxo-6(1)-substituted benzyl-4(1),5(1) dihydro-1(1),2(1),4(1)-triazin-3-yl)mercaptoacetyl-3-arylsydnones by the reaction of 3-aryl-4-bromoacetylsydnones with 6-substituted-4-amino-3-mercapto-1,2,4 triazin-5-ones is described. The IR, (1)H NMR, mass spectra and elemental analysis characterized the newly synthesized compounds. The synthesized compounds were screened for their antimicrobial activity. All the compounds showed higher activity than that of standard drug during antimicrobial studies and the activity was comparable with the standard drug for antifungal activity. PMID- 18387711 TI - Lanostane-type triterpenoids from the sclerotia of Inonotus obliquus possessing anti-tumor promoting activity. AB - Two new lanostane-type triterpenoids, 1 and 2 besides two known lanostane-type triterpenoids, 3 and 4 were isolated from the sclerotia of Inonotus obliquus. Their structures were determined to be lanosta-8,23E-diene-3beta,22R,25-triol (1) and lanosta-7:9(11),23E-triene-3beta,22R,25-triol (2) by spectral data. These compounds were tested for their anti-tumor-promoting activity using a short-term in vitro assay for EBV-EA activation induced by TPA. Compounds 1, 2 and 4 were stronger than the positive control, oleanolic acid. The most abundant compound 4 was investigated for the inhibitory effect in a two-stage carcinogenesis test on mouse skin using DMBA as an initiator and TPA as a promoter. Compound 4 was found to exhibit the potent anti-tumor promoting activity in the in vivo carcinogenesis test. PMID- 18387712 TI - [From inappropriateness of hospital use to an irrelevant medicine]. PMID- 18387713 TI - [Hypocomplementemic urticarial vasculitis]. AB - Hypocomplementemic urticarial vasculitis is a rare disorder characterized by the presence of C1q precipitins associated with a syndrome of urticarial vasculitis, arthralgias, ocular inflammation and obstructive-lung disease. We report the case of a 48-year-old woman with hypocomplementemic urticarial vasculitis. Because of dependance to corticosteroids, cyclophosphamide-pulse therapy was started and resulted in significant clinical improvement. Mycophenolate mofetil was effective as maintenance therapy and resulted in complete resolution of rash, arthralgias, arthritis and uveitis, but had no effect on the obstructive-lung disease. PMID- 18387714 TI - [Munchausen syndrome with forgery on biologic results. A case report]. AB - Munchausen syndrome is a disorder defined by the following: acute factitious symptoms leading to inappropriate investigation and therapy, a restless journey from hospital to hospital and autobiographical falsification. We report here a 20 year-old woman who presented at our hospital consultation of internal medicine with laboratory-test results suggesting the diagnosis of leukemia. A new complete blood cells count and a medullogram by sternal puncture did not show any abnormality. Comparative examination of laboratory-test sheets lead to the diagnosis of Munchausen syndrome as some results had been falsified. With unlimited access to information through internet and word or image processing softwares, laboratory results have become easy to falsify nowadays, particularly for patients with Munchausen syndrome, who may then be quite difficult to diagnose accurately in the context of medical consultation. PMID- 18387715 TI - [Muscle lymphoma: a case report]. AB - Skeletal muscle involvement is uncommon in lymphoma, occurring in less than 1.5% of patients. We report the original case of a 61-year-old man who presented with pseudotumoral muscle lesions of the lower limbs, revealing non-Hodgkin T-cell lymphoma. In our patient, magnetic resonance imaging (MRI) was useful in clearly revealing the detailed anatomic extent of muscle change; indeed, MRI showed muscle enhancement after intravenous administration of gadolinium on T1-weighted images as well as high-signal intensity on T2-weighted images. Moreover, MRI was helpful in guiding the optimal site for muscle biopsy. PMID- 18387716 TI - [Abdominal pain]. PMID- 18387717 TI - [Flexible intensive insulin therapy: a change in mentalities]. PMID- 18387718 TI - Diagnosis and treatment of (disease-related) in-hospital malnutrition: the performance of medical and nursing staff. AB - BACKGROUND & AIMS: Malnutrition continues to be an important problem in health care which is still under recognized and underrated in developed countries. This study aims to describe current practice in diagnosing and treating malnutrition by medical doctors, medical students and nurses prior, during and after hospitalisation. METHODS: Prospective analysis of current practice in assessing nutritional status and prescribing treatment by medical and nursing staff in a cohort of hospitalised patients from the general medical wards of the VU University Medical Center, Amsterdam. Comparison of objective identification of malnutrition by an independent observer with subjective identification by the medical and nursing staff. Quantification of diagnosing, treating and communicating malnutrition before, during and following hospital stay by medical doctors, medical students and nurses by evaluating the written information in medical and nursing charts, and referral and discharge letters. RESULTS: Three hundred and ninety-five women and men, aged 19-96 years, were included from June to September 2005. The prevalence of malnutrition was 31.9%. Nutritional information was not mentioned in written referrals. Medical doctors performed nutritional assessment in 15.3%, medical students in 52.8%, and nurses in 29.9% of their patients. Medical doctors were the most capable of differentiating between malnourished and well-nourished patients as a basis for undertaking nutritional assessment, although this was still inadequate. Little nutritional intervention was applied during hospital stay. Information on nutritional status was lacking in most discharge letters. Nutritional follow-up was appointed in 1.2%. CONCLUSIONS: Nutritional assessment and intervention were not sufficiently applied by any professional at any stage of the pre-, actual and post hospitalisation period. PMID- 18387720 TI - A nasal vaccine comprising B-subunit derivative of Shiga toxin 2 for cross protection against Shiga toxin types 1 and 2. AB - Enterohemorrhagic Escherichia coli (EHEC) produces Stx1 and Stx2 causing severe diseases. Their B-subunits (StxBs) exhibit low immunogenicity and the anti-StxB antibodies neutralizing both Stxs has not been prepared yet. By intranasal vaccination with His-tagged-StxB (Stx1B-His or Stx2B-His) plus a mutant heat labile enterotoxin (mLT) in mice, their serum and lung fluid reacted with appropriate StxB. Mice vaccinated with Stx2B-His plus mLT had antibodies reacting Stx1B and showed the resistance to toxemia of Stx1 and Stx2. This is the first demonstration to get anti-Stx2B serum neutralizing both Stxs. These suggest that the nasal vaccination with Stx2B-His and mLT is effective for preventing toxemias by EHEC. PMID- 18387719 TI - Phase 2a trial of 0, 1, and 3 month and 0, 7, and 28 day immunization schedules of malaria vaccine RTS,S/AS02 in malaria-naive adults at the Walter Reed Army Institute of Research. AB - BACKGROUND: Immunization with RTS,S/AS02 consistently protects some vaccinees against malaria infection in experimental challenges and in field trials. A brief immunization schedule against falciparum malaria would be compatible with the Expanded Programme on Immunization, or in combination with other prevention measures, interrupt epidemic malaria or protect individuals upon sudden travel to an endemic area. METHODS: We conducted an open label, Phase 2a trial of two different full dose schedules of RTS,S/AS02 in 40 healthy malaria-naive adults. Cohort 1 (n=20) was immunized on a 0, 1, and 3 month schedule and Cohort 2 (n=20) on a 0, 7, and 28 day schedule. Three weeks later, 38 vaccinees and 12 unimmunized infectivity controls underwent malaria challenge. RESULTS: Both regimens had a good safety and tolerability profile. Peak GMCs of antibody to the circumsporozoite protein (CSP) were similar in Cohort 1 (78 microg/mL; 95% CI: 45 134) and Cohort 2 (65 microg/mL; 95% CI: 40-104). Vaccine efficacy for Cohort 1 was 45% (95% CI: 18-62%) and for Cohort 2, 39% (95% CI: 11-56%). Protected volunteers had a higher GMC of anti-CSP antibody (114 microg/mL) than did volunteers with a 2-day delay (70 microg/mL) or no delay (30 microg/mL) in the time to onset of parasitemia (Kruskal-Wallis, p=0.019). A trend was seen for higher CSP-specific IFN-gamma responses in PBMC from protected volunteers only in Cohort 1, but not in Cohort 2, for ex vivo and for cultured ELISPOT assays. CONCLUSION: In malaria-naive adults, the efficacy of three-dose RTS,S/AS02 regimens on either a 0, 1, and 3 month schedule or an abbreviated 0, 7, and 28 day schedule was not discernibly different from two previously reported trials of two-dose regimens given at 0, 1 month that conferred 47% (95% CI: -19 to 76%) protection and in another trial 42% (95% CI: 5-63%). A strong association of CSP specific antibody with protection against malaria challenge is observed and confirms similar observations made in other studies. Subsequent trials of adjuvanted RTS,S in African children and infants on a 0, 1, and 2 month schedule have demonstrated a favorable safety and efficacy profile. PMID- 18387722 TI - Carcinoma of the midline dorsum of the tongue. AB - We present a case of an 80 year old female who had previously been diagnosed with lichen planus of the tongue dorsum some 10 years previously. Due to a change in the appearance of the lesion a biopsy revealed squamous cell carcinoma of the tongue. We discuss this rare diagnosis in conjunction with the possibility of an association of squamous cell carcinoma and lichen planus. PMID- 18387721 TI - Protective effects of recombinant staphylococcal enterotoxin type C mutant vaccine against experimental bovine infection by a strain of Staphylococcus aureus isolated from subclinical mastitis in dairy cattle. AB - Staphylococcus aureus is one of the main etiological agents of bovine mastitis; however, antibiotics that are effective against bovine strains of S. aureus are not currently available. Staphylococcal enterotoxin type C (SEC), a superantigen, is the enterotoxin most frequently expressed by bovine strains of S. aureus and one of immunogenic determinants. The purpose of this study was to evaluate the protective effectiveness of recombinant SEC mutant vaccine (MastaVactrade mark) against experimentally induced bovine infection. Three representative SEC secreting strains were selected from 9 candidate isolates that showed various intensities of pathogenicity on mice and inoculated into 5 lactating dairy cattle at a concentration of 50-5.0x10(8) CFU per quarter. The optimal experimental bovine subclinical mastitis model was produced by inoculation with 50 CFU of S. aureus 409 per quarter, a level which was not lethal to mice. After the experimental model was determined, other 3 cattle were intramuscularly administered three doses of vaccine at day 0, at 2 wks and at 6 wks. Nine quarters of 3 vaccinated cattle and 8 quarters of 3 control cattle were then challenged with S. aureus 409. An SEC-specific ELISA test conducted at 4 wks post immunization confirmed the presence of a high antibody titer against SEC in all vaccinated cattle. The somatic cell counts from the vaccinated group remained relatively low, whereas those of control group increased significantly after challenge with S. aureus. After challenge, S. aureus was not isolated from any cattle in the vaccinated group, whereas it was isolated from 75% of the cattle in the control group. These results indicate that recombinant SEC mutant vaccine had a protective effect against S. aureus intramammary infection in lactating cattle. PMID- 18387723 TI - The interface between research and policy: experience from South Africa. AB - Increasing attention has been paid in recent years to efforts to strengthen the impact of research on policy in low- and middle-income countries. However, the processes by which such research might have policy impact remain a subject of debate. This paper presents an analysis of the research/policy interface, drawing on the experiences of two South African health policy and systems research (HPSR) units and one specific study which traced the development and implementation of three areas of health care financing policy change and debate between 1994 and 1999. The analysis is based primarily on the authors' own experiences and has been developed through a deliberate process of reflection. It suggests, first, that it is important to acknowledge the conceptual and symbolic uses and impacts of research--perhaps, particularly in relation to the system-oriented work of HPSR groups. These uses may not be verifiable by specific changes in policy and practice but are important contributions to the policy environment and do filter into policy-makers' understandings and actions. Second, achieving any form of impact on policy is linked to the attention researchers pay to the context in which the research is undertaken, the nature and credibility of the research; and the importance of nesting any single project in a broader programme of engagement with the policy environment that builds trust in the researchers. PMID- 18387724 TI - Risperidone-induced double incontinence. PMID- 18387725 TI - [Nipple and areola reconstruction by tattooing, "F" and "Z" flaps]. AB - Nipple and areola reconstruction is very important in the evaluation of the quality of breast reconstruction. It can be done during the primary or secondary breast reconstruction or later. We have performed the techniques of nipple reconstruction routinely since 1992. Under local anesthesia during a second operative time or general anesthesia during breast reconstruction, the local "F" and "Z" skin flaps and tattooing grant a quality result in the wound and the long term projection. They are easily reproduced, rapid and as there is no graft the choice of the incisions grants a good tolerance. Complications are rare and it is always possible to use other techniques in case of poor result. We also present the main techniques of nipple and areola reconstruction with their advantages and limits. PMID- 18387726 TI - [Partial reconstruction after conservative treatment for breast cancer: classification of sequelae and treatment options]. AB - Most patients presenting with breast cancer are treated by breast conserving treatment (BCT). Some of these patients present with poor cosmetic results and ask for partial breast reconstruction. These reconstructions following BCT are presenting more frequently to plastic surgeons as a difficult management problem. We have defined and published a classification of the different cosmetic sequelae (CS) after BCT into three types. This classification helps to analyse these complex deformities aggravated by radiotherapy. Furthermore, our classification helps to choose between the different surgical techniques and propose the optimal option for their surgical correction. Our initial publications reported 35 and 85 patients: we have currently operated more than 150 cases of CS after BCT. Type-1 CS are defined by an asymmetry between the two breasts, with no distortion or deformity of the radiated breast. Type-2 CS are those with an obvious breast deformity, that can be corrected with a partial reconstruction of the breast. Type-3 CS are those with such a deformity that only a mastectomy with total reconstruction of the breast can be performed. Most of the patients present with type-2 CS, but are reluctant to undergo what they feel is a major reconstructive procedure: in a initial prospective series of 85 patients operated for CS after BCT, 48 (56.5%) had type-1 CS, 33 patients (38.8%) type-2 CS and four patients (4.7%) type-3 CS. Type-1 patients should be managed essentially by contralateral symmetrizing procedures. One should limit any surgery on the radiated breast, as a mammoplasty or an augmentation is at high risk of complications. Type-2 is the most difficult to manage and requires all the surgical armamentarium of breast reconstructive surgery. The insetting of a myocutaneous flap is often necessary and autologous fat grafting is a promising tool in selected cases. Type-3 CS requires mastectomy and immediate reconstruction with a myocutaneous flap. The major development though in the past 10 years has been the development of oncoplastic techniques at the time of the original tumour removal, in order to avoid most of type 2 and type 3 deformities. This paper reaffirms the validity of the Cosmetic Sequelae classification as a simple, practical guide for breast reconstructive surgeons. It discusses the various choices of reconstructive procedures available, the importance of "preventing" these CS and defining the role of the plastic surgeon in the management of these patients. PMID- 18387727 TI - Inhibition of glycogen synthase kinase-3 reduces L-DOPA-induced neurotoxicity. AB - The neurotoxicity of l-3,4-dihydroxyphenylalanine (L-DOPA), used for the treatment of Parkinson's disease, remains controversial. Although there are many reports suggesting that long-term treatment of L-DOPA causes neuronal death, an increasing body of recent evidence has proposed that L-DOPA might be neuroprotective rather than neurotoxic. We investigated the effect of L-DOPA on neuronally differentiated PC12 (nPC12) cells by treating cells with various concentrations of L-DOPA for 24h. We also studied whether glycogen synthase kinase (GSK)-3 activation is related to L-DOPA-induced neurotoxicity by simultaneously treating cells with several concentrations of L-DOPA and a GSK-3 inhibitor for 24h. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay, trypan blue staining, cell counting kit-8, and DAPI staining all showed that L-DOPA decreased nPC12 cell viability at high concentrations. In addition, 100 microM L-DOPA treatment significantly increased the activity of GSK 3 and death signals including cytochrome c, activated caspase-3 and cleaved PARP, and decreased survival signals including heat shock transcription factor-1 in a concentration-dependent manner. Treatment with GSK-3 inhibitor VIII or lithium chloride prevented L-DOPA-induced cell death. Together, these results suggest that L-DOPA induces neuronal cell death at high concentrations and that the neurotoxic effect of L-DOPA might be mediated in part by GSK-3 activation. PMID- 18387728 TI - Comparative study in desensitizing efficacy using the GaAlAs laser and dentin bonding agent. AB - OBJECTIVE: To compare the clinical efficacy of the GaAlAs laser and dentin bonding agent in treating dentin hypersensitivity. METHODS: One hundred and forty teeth from 70 patients diagnosed with dentin hypersensitivity, were divided into two groups: In group I, teeth were irradiated with 30 mW GaAlAs laser for 1 min and in group II, teeth were applied with dentin bonding agent. Sensitivity was assessed by tactile and thermal tests, measured with the criteria proposed by Uchida at baseline and after treatment; immediately, at 15 and at 30 days. RESULTS: The reduction of dentin hypersensitivity was observed in both treatments using the GaAlAs laser and dentin bonding agent. However, a greater reduction was observed over time up to day 15 and no significant change was observed between days 15 and 30. Statistically significant differences in the level of dentin hypersensitivity between bonding agent and the GaAlAs laser were found at each observation period (p < 0.05). CONCLUSION: The GaAlAs laser had less desensitizing efficacy compared with dentin bonding agent. However, the desensitizing effect of the GaAlAs laser still could be considered an effective therapy for treating dentin hypersensitivity. PMID- 18387729 TI - An unusual case of clitoromegaly. PMID- 18387730 TI - Land cover mapping of wetland areas in an agricultural landscape using SAR and Landsat imagery. AB - Saline wetlands in the Monegros Desert, NE Spain, are situated in an agricultural landscape which is undergoing significant changes. Agricultural intensification in recent decades and current installation of new irrigation systems threaten these valuable habitats, set to be included in the Natura2000 network. Their preservation and successful management depend on the information available regarding the transformation of surrounding areas. When soil and vegetation maps at adequate scale are not available, remote sensing is an alternative means to obtain needed data. We have used SAR data, taking advantage of the soil surface characteristics perceived in SAR images. The objective of this work is to explore the capability of multitemporal SAR data to characterize the land covers of these wetlands and their environment. We have developed specific contextual classifications which take into account the statistical properties of the radar distribution. Moreover, we tested the contribution of radar in Landsat classification. PMID- 18387731 TI - Laser fibre deterioration and loss of power output during photo-selective 80-w potassium-titanyl-phosphate laser vaporisation of the prostate. AB - BACKGROUND: The potassium-titanyl-phosphate (KTP) laser technique for photo selective vaporisation of the prostate (PVP) has been regularly improved over the last decade. Nonetheless, decreasing efficiency of tissue vaporisation during the course of the operation and macroscopic alterations of the laser fibre's tip are regularly observed and seem to affect the outcome of this procedure. OBJECTIVE: To investigate the course of power output and to determine the type and extent of fibre deterioration during PVP. DESIGN, SETTING, AND PARTICIPANTS: Forty laser fibres were investigated during PVP in 35 consecutive patients with prostatic bladder outflow obstruction between January 2007 and August 2007 in a university hospital. INTERVENTION: All patients underwent PVP performed by three different surgeons using the 80-W KTP laser. MEASUREMENTS: Power output was measured at the beginning and regularly throughout PVP and throughout in vitro vaporisation without fibre-tissue contact. Microscopic documentation of the fibre tip was performed after the procedure. RESULTS AND LIMITATIONS: Carbonisation and melting of the fibre tip was regularly visible and appeared to be more pronounced as more energy was applied. Additionally, 90% of the fibres showed a significant decrease of power output during PVP, resulting in an end-of-lifespan (ie, 275-kilojoule) median power output of 20% of the initial value. Final median power output after in vitro vaporisation was 83% of the starting value. The extent of the structural and functional changes might only be valid for the operative technique performed in this investigation. CONCLUSIONS: Fibre deterioration caused significant reduction of power output during PVP. This finding is an explanation for the often observed decreasing efficiency of tissue ablation and may also be responsible for some of the typical drawbacks and complications of PVP. Hence, improvements in fibre quality are necessary to advance the efficiency of this technique. PMID- 18387732 TI - Editorial comment on: Laser fibre deterioration and loss of power output during photo-selective 80-W potassium-titanyl-phosphate laser vaporisation of the prostate. PMID- 18387733 TI - The impact of neuroendoscopy in the emergency setting--a retrospective study of imaging, intraoperative findings, and surgical outcome in 55 patients. AB - OBJECTIVES: To assess whether neuroendoscopy is an improved and effective diagnostic as well as therapeutic tool in the emergency setting compared with neuroimaging studies. PATIENTS AND METHODS: Sixty-two preoperative computed tomography (CT) and magnetic resonance (MR) scans from 55 adult patients admitted in the emergency ward were compared with equal number of neuroendoscopy observations by independent observers, who included cases of raised intracranial pressure related to hydrocephalus, shunt dysfunction, intracranial cysts, brain tumours, subdural effusions, etc. RESULTS: According to the results obtained, three groups were formed. Postoperative findings matched CT/MR images in 38.7% of all the procedures (Group 1). In 25.8%, neuroimages were different from endoscopy findings, nevertheless the differences did not justify any modification in the original surgical approach (Group 2). Finally, differences between endoscopical and preoperative studies in 34.48% of the procedures did justify modifications in the surgical plan (Group 3). Extensive fibrosis within the ventricular system associated with poor anatomic landmark visibility was found to be the main cause of surgical modification in the majority of these cases. CONCLUSIONS: Usefulness of neuroendoscopy to increase the accuracy of intraoperative diagnosis, and therefore modify the surgical procedure in an emergency setting, is stressed. PMID- 18387734 TI - Geochemical modelling of arsenic and selenium leaching in alkaline water treatment sludge from the production of non-ferrous metals. AB - Geochemical modelling of leaching of oxyanion forming elements such as arsenic (As) and selenium (Se) is frequently not successful. A consistent thermodynamic dataset of As and Se was therefore composed, not only including precipitation, but also adsorption and solid solution, and was applied to the pH-dependent leaching behaviour of As and Se in an alkaline residue with a pH 11.1 from the lime treatment of sulphuric acid wastewaters from the production of non-ferrous metals. The As and Se content ranged up to 6.7 wt% and 0.29 wt%, respectively and speciation analysis showed that 96.3% of As occured as arsenate whereas Se speciation comprised 79% selenate and 21.0% selenite. XRD and SEM/EDX analysis showed that arsenate occurred as rauenthalite (Ca(3)(AsO(4))(2).10H(2)O), associated with gypsum, the most important mineral. Arsenate and arsenite concentrations were only slightly below equilibrium with rauenthalite and calciumarsenite (CaHAsO(3)), respectively and consideration of adsorption and solid solution only marginally improved model predictions. Selenate (Se(VI)) and selenite (Se(IV)), on the other hand, were far from equilibrium with their corresponding calcium metalate. The application of solid solutions and adsorption of Se(VI) and Se(IV) oxyanions with gypsum, calcite and ettringite significantly improved model predictions but missing thermodynamic data and especially the lack of a comprehensive model for solid solution and surface exchange with calcite and ettringite still hampered efficient modelling. PMID- 18387735 TI - Malachite green adsorption by rattan sawdust: isotherm, kinetic and mechanism modeling. AB - In this work, the adsorption of malachite green (MG) on rattan sawdust (RSD) was studied at 30 degrees C. The results indicated that RSD can be used as a low-cost adsorbent for the removal of MG dye from aqueous solutions. Equilibrium data were analyzed by two isotherms, namely the Freundlich isotherm and the Langmuir isotherm. The best fit to the data was obtained with the Langmuir isotherm. The monolayer adsorption capacity of RSD was found to be 62.71 mg/g. The adsorption kinetics can be predicted by the pseudo-first-order model. The mechanism of adsorption was also studied. It was found that for a short time period the rate of adsorption is controlled by film diffusion. However, at longer adsorption times, pore-diffusion controls the rate of adsorption. The amount adsorbed on the outer surface was estimated from the time where film-diffusion stops controlling the adsorption rate. PMID- 18387736 TI - Simultaneous adsorption of phenol and cadmium on amphoteric modified soil. AB - Surface modification is an effective way to enhance adsorption of pollutants by soil. In this study, we investigated the individual adsorption of cadmium ion (Cd(2+)) and phenol and also in combination by the clay layer of a loessial soil treated with the amphoteric modifier, duodalkylbetaine (BS-12). Three levels of BS-12 modification were compared in this experiment: (1) unmodified soil (CK), (2) modification with an amount of BS-12 equivalent to 50% of the soil's CEC (50BS) and (3) modification with an amount of BS-12 equivalent to 100% of the soil's CEC (100BS). Cd(2+) adsorption was 0.92-1.70 times higher in the amphoteric modified soil compared to unmodified soil. Adsorption isotherms for Cd(2+) displayed a L1-type shape. Phenol adsorption was 1.25-4.35 times higher in the amphoteric modified soil compared to the unmodified control. The adsorption isotherms of phenol on amphoteric modified soils were generally linear, but changed to L1-type isotherms for modified soil in the Cd(2+)+phenol treatment at 40 degrees C. The results clearly showed that amphoteric modified soil had the ability to simultaneously adsorb Cd(2+) and phenol. Cd(2+) adsorption by the amphoteric modified soil was related to the initial concentration of Cd(2+) in the supernatant. Cd(2+) adsorption in the 100BS treatment exceeded adsorption in the 50BS treatment when Cd(2+) initial concentrations were higher than approximate 200 microg mL(-1). Phenol adsorption by modified soils decreased in the order: 100BS>50BS>CK and was primarily determined by the surface hydrophobicity of the soil. For the unmodified soil, total adsorption in the Cd(2+)+phenol treatment was slightly lower compared to treatments that contained only Cd(2+) or phenol. This indicated an antagonistic effect between the adsorption of Cd(2+) and phenol, which was reduced after amphoteric modification. A comparison of temperature effects on Cd(2+) and phenol adsorption indicated that Cd(2+) was both physically and chemically adsorbed by the amphoteric modified soil, but phenol was primarily adsorbed physically. PMID- 18387737 TI - Color removal from textile dyebath effluents in a zeolite fixed bed reactor: determination of optimum process conditions using Taguchi method. AB - Taguchi method was applied as an experimental design to determine optimum conditions for color removal from textile dyebath house effluents in a zeolite fixed bed reactor. After the parameters were determined to treat real textile wastewater, adsorption experiments were carried out. The breakthrough curves for adsorption studies were constructed under different conditions by plotting the normalized effluent color intensity (C/C(0)) versus time (min) or bed volumes (BV). The chosen experimental parameters and their ranges are: HTAB concentration (C(htab)), 1-7.5 gL(-1); HTAB feeding flowrate (Q(htab)), 0.015-0.075 L min(-1); textile wastewater flowrate (Q(dye)), 0.025-0.050 L min(-1) and zeolite bed height (H(bed)), 25-50 cm, respectively. Mixed orthogonal array L(16) (4(2)x2(2)) for experimental plan and the larger the better response category were selected to determine the optimum conditions. The optimum conditions were found to be as follows: HTAB concentration (C(htab))=1g L(-1), HTAB feeding flowrate (Q(htab))=0.015 L min(-1), textile wastewater flowrate (Q(dye))=0.025 L min(-1) and bed height (H(bed))=50 cm. Under these conditions, the treated wastewater volume reached a maximum while the bed volumes (BV) were about 217. While HTAB concentration, gL(-1) (A); zeolite bed height, cm (D) and wastewater flowrate, L min(-1) (C) were found to be significant parameters, respectively, whereas, HTAB flowrate, L min(-1) (B) was found to be an insignificant parameter. PMID- 18387739 TI - The chemical and biological characteristics of coke-oven wastewater by ozonation. AB - A bench-scale bubble column reactor was used to investigate the biological and chemical characteristics of coke-oven wastewater after ozonation treatment through the examination of selected parameters. Color and thiocyanate could be removed almost entirely; however, organic matter and cyanide could not, due to the inadequate oxidation ability of ozone to remove ozonated byproducts under given experimental conditions. The removal of cyanide and total organic carbon were pH-dependent and were found to be efficient under neutral to alkaline conditions. The removal rate for thiocyanate was about five times that of cyanide. The ozone consumption ratio approached to about 1 at the early stage of ozonation (time <20 min), indicating that easily degraded matter was present, and mostly ozone was used to oxidize the pollutants. As ozonation progressed, the consumption ratio decreased to 0.2, and TOC removal (eta(TOC)) increased to 30%, indicating that easily degraded pollutants were degraded almost entirely. The effect of ozonation on the subsequent biological treatment unit (i.e., activated sludge process) was determined by observing the ratio of 5-day biological oxygen demand to chemical oxygen demand (BOD(5)/COD) and the specific oxygen utilization rate (SOUR). The results indicated that the contribution of ozonation to inhibition reduction was very significant but limited to the enhancement of biodegradation. The operation for ozonation of coke-oven wastewater was feasible under neutral condition and short ozone contact time in order to achieve better performance and cost savings. PMID- 18387738 TI - In situ stabilization of chromium(VI) in polluted soils using organic ligands: the role of galacturonic, glucuronic and alginic acids. AB - Laboratory batch sorption and column experiments were performed to investigate the role of organic ligands such as galacturonic, glucuronic and alginic acids (main constituents of bacterial exopolymeric substances (EPS)) on Cr(VI) uptake and transport in heterogeneous subsurface media. Our batch sorption experiments demonstrate the addition of galacturonic, glucuronic and alginic acids to soils enhances Cr(VI) uptake by soil at pH values <7.7 depending on the concentration of the ligand and pH used. The enhanced Cr(VI) uptake at pH values <7.7 may be explained through either the catalytic reduction of Cr(VI) to Cr(III) by the surface-bound organic matter/Fe oxides and/or the dissolved metal ions (e.g., Fe(III)) from the soil. On the other hand, organic ligands have no or little effect on Cr(VI) uptake under highly alkaline pH conditions since the catalytic Cr(VI) reduction decreases with increasing pH. Similarly, the results from column experiments show that, depending on the concentration of organic ligands, the Cr(VI) breakthrough curves were significantly retarded relative to the organic acid-free systems at pH 7.6. A significant portion of Cr(VI) initially added to the feed solution was not readily recoverable in the effluent, indicating Cr(VI) reduction in columns, most probably catalyzed by surface-bound metal-oxides (e.g., Fe oxides) or dissolved metal ions such as Fe(II; III). The overall results suggest that EPS constituents such as glucuronic, galacturonic and alginic acids may play a significant role on Cr(VI) stabilization in subsurface systems under acidic to slightly alkaline pH conditions. PMID- 18387740 TI - Serotonin receptor 1A-1019C/G variant: impact on antidepressant pharmacoresponse in melancholic depression? AB - Previous studies on the effects of serotonin receptor 1A (5-HT1A) gene variation on treatment response in depression revealed inconsistent results with studies pointing towards a detrimental influence of the 5-HT1A-1019G allele on antidepressant treatment response, while others did not discern any involvement of 5-HT1A variants. In order to further delineate the impact of 5-HT1A gene variation on pharmacoresponse in depression over 6 weeks of antidepressant treatment, the influence of the 5-HT1A-1019C/G (rs6295) polymorphism was investigated in 340 Caucasian patients with a Major Depressive Episode (DSM-IV) with particular attention to the subtype of depression (major depression and melancholic depression). Antidepressant treatment response across 5-HT1A-1019C/G genotype groups showed no differences in either Major Depressive Episode or major depression between genotype groups, whereas stratification for the melancholic subtype of depression revealed a significantly worse treatment response as conferred by the -1019CC genotype (p=0.02). The poorer treatment response in melancholic depression could first be detected in week 2 (p=0.03), continuing until week 6 and showing a maximum effect in week 3 (p=0.01). The present study adds to the clarification of the role of 5-HT1A variation in treatment response in major depression by providing preliminary support for poor treatment response mediated by the 5-HT1A-1019C allele repressing 5-HT1A activity specifically in the melancholic subtype of depression. PMID- 18387741 TI - Association study of a functional catechol-O-methyltransferase polymorphism and executive function in elderly males without dementia. AB - Cognitive function in older people is a major factor influencing quality of life. The catechol-O-methyltransferase (COMT) gene, which is essential in the metabolic degradation of prefrontal dopamine, has been considered as a leading candidate gene in the variation in cognitive performance. The aim of this study was to investigate the effect of a functional COMT (Val158Met) polymorphism on several cognition domains in a relatively homogeneous population consisting of elderly Chinese males without dementia. Six neuropsychological measurements, including Spatial Span Forward and Backward, Digit Span Forward and Backward, and Trail Making Test-A and -B, were assessed in 161 aged males. It was found that the Met/Met carriers showed a better performance than the Val/Met and Val/Val subjects on the Digit Span Forward (a measure of general attention; p=0.017, after correction for education level) test, but not on the other cognitive tests. These findings suggest that the COMT Val158Met genotype may contribute to differences in normal cognitive aging, particularly in the area of general attention. PMID- 18387742 TI - Cerebral metabolic effects of fluoxetine, fluvoxamine, paroxetine and sertraline in the conscious rat. AB - The selective serotonin reuptake inhibitors (SSRI) exert a wide range of neurochemical and therapeutic activities. To investigate the neural effectors of SSRIs, we measured the regional cerebral metabolic rates for glucose (rCMRglc) in 56 brain regions of Fischer-344 rats 30 min after intraperitoneal injection of 0.4, 4 or 40 mg/kg of fluoxetine or fluvoxamine or after 4 mg/kg of paroxetine or sertraline. Both shared and drug-specific effects were detected. While all four SSRIs similarly reduced rCMRglc in a network of subcortical brain regions including the amygdala, locus coeruleus, basal ganglia and hypothalamic paraventricular nuclei, fluvoxamine, paroxetine and sertraline reduced rCMRglc also in the hippocampus and sertraline in the lateral habenula. The topography and the relation to dose of rCMRglc reductions by SSRIs differ from those of other classes of antidepressants, thus suggesting that SSRIs may specifically modulate brain areas involved in the physiological responses to stress. PMID- 18387743 TI - A novel technique for identification of Ascaris suum cohorts in pigs. AB - The objective of the present study was to develop a fast, cheap and reliable technique for identifying different cohorts of the swine parasite, Ascaris suum. A polymerase chain reaction linked restriction fragment length polymorphism (PCR RFLP) technique on mt-DNA was used to identify unique haplotypes of four gravid A. suum females on agarose gels after eggs were recovered from each of the worms. Each of four pigs was inoculated with 2000 embryonated eggs originating from one of the four identified Ascaris haplotypes, respectively. Ascaris larvae were isolated from the small intestine at day 14 post-infection using an agar technique. Single larvae from each pig were transferred to 96-well PCR plates and a simple DNA extraction using a worm lysis buffer was carried out and followed by the PCR-RFLP analysis. More than 100 larvae from each of the four pigs were analysed and all were found to have the same haplotype as the parental female. We conclude that unique haplotypes of female A. suum and offspring can be identified by means of PCR-RFLP on mt-DNA and suggest that this method can be used in future research on Ascaris population biology using cohorts with distinct mt-DNA profile. PMID- 18387744 TI - Infectivity of Hepatozoon americanum cystozoites for a dog. AB - Hepatozoon americanum cystozoites from experimentally infected, laboratory-raised rodents were fed to a Hepatozoon-free dog. Gamonts were detected by examination of blood smear 42 and 56 days post-exposure. PCR analysis of blood was positive for the 18S rRNA Hepatozoon gene on days gamonts were demonstrated. Meronts were detected histologically in a skeletal muscle biopsy 90 days after ingestion of cystozoites. Sequencing confirmed that the parasite in the dog was H. americanum. Xenodiagnosis was conducted by replete feeding of Ambylomma maculatum larvae on the dog; 40 days after detachment, sporulated oocysts were recovered from recently molted nymphs. PMID- 18387746 TI - Genomic analyses of Haemonchus contortus infection in sheep: abomasal fistulation and two Haemonchus strains do not substantially confound host gene expression in microarrays. AB - To determine whether fistulation and differing strains of Haemonchus contortus complicate genome analysis of the host response to infection, two pilot experiments examined parasite development and gene expression in the abomasal mucosa of parasitised sheep. No significant differentially-expressed genes were detected in a comparison between ivermectin-susceptible McMaster and ivermectin resistant CAVR strains of H. contortus. This demonstrated that the sheep response was not significantly altered by the ivermectin-resistance status of the parasite. However, sheep infected with McMaster strain had a significantly lower proportion of larvae and a higher mean FEC at post-mortem than sheep infected with CAVR, suggesting that McMaster larvae advance to patency faster than CAVR larvae. Abomasal fistulation resulted in significant upregulation of three genes and significant downregulation of two genes. Fistulated sheep had significantly lower FEC than the other groups but the proportion of larvae at post-mortem was not significantly different to other groups infected with the same strain (CAVR). Hence fistulation does not alter establishment of the CAVR isolate, but may slow its progression to patency. The observation that different H. contortus strains and abomasal fistulation induced minimal changes in mucosal gene expression validated the design of a subsequent experiment (manuscript in preparation) where sequential biopsies taken during infection were analysed by microarray to describe the molecular responses which inhibit larval establishment. PMID- 18387745 TI - Detection of Assemblage A, Giardia duodenalis and Eimeria spp. in alpacas on two Maryland farms. AB - Sixty-one fecal samples were collected from adult alpacas and crias (ages 10 weeks to 10 years) on two farms in central Maryland. The farms raised both suri (silky-haired) and huacaya (crimpy-haired) breeds. Females and crias were housed together on pasture, whereas older/breeding males were maintained on separate pastures. Samples were subjected to a density gradient centrifugation protocol to concentrate parasites and remove fecal debris and were examined by immuno fluorescent and differential interference contrast microscopy. Oocysts of Eimeria spp. were noted in 14 fecal samples, 6 on MD-1 and 8 on MD-2. Based on oocyst morphometrics two species of Eimeria were present: E. punoensis (19.2 microm x 16.5 microm) and E. alpacae (23.7 microm x 19.5 microm). Five animals shed exclusively E. punoensis, seven shed exclusively E. alpacae, and two had mixed infections. The Eimeria infections were not associated with obvious clinical signs. To determine the presence of Cryptosporidium and Giardia species and genotypes, DNA was extracted from feces and subjected to PCR utilizing specific primers for the ssu-rRNA gene for both parasites. All PCR positive samples were further analyzed by DNA sequencing to identify the species or genotypes that were present. Assemblage A, G. duodenalis was detected in fecal samples from two alpacas on MD-1 and in one alpaca on MD-2. Assemblage E, G. duodenalis and Cryptosporidium spp. were not detected on either farm. Although the prevalence on these two farms was low, alpacas can harbor zoonotic G. duodenalis, and this should be borne in mind by persons interacting with the animals. PMID- 18387747 TI - Prevalence of anti-T. canis antibodies in stray dogs in Mexico City. AB - Toxocara canis is a common intestinal helminth found in dogs. In humans, it is a cause of Visceral Larva Migrans (VLM), a zoonosis rarely studied in Mexico. The aim of this study is to examine, by means of the indirect haemaglutination test (IHAT), the prevalence of antibodies of T. canis in the serum of stray dogs in Mexico City. METHODS AND MATERIALS: 141 stray dog serum samples from three different districts of the city were analyzed: Iztacalco (49), Iztapalapa (49) and Coyoacan (43). In each location three study groups were formed. Group I with 35 dogs (less than a year old), Group II with 91 dogs (ages 113% nucleotide divergence) led to specify the classification of this virus family. Phylogenetic analysis of partial sequences of V9-related isolates combined with Erythrovirus sequences in sequences banks indicates an organization into three well-individualized genotypes. Analysis of the nearly full-length genome sequences show an ancient separation between the three genotypes lineages. Genotype 3 (the most ancient lineage) could have originated in Africa. The functional regions of major proteins are conserved in the three genotypes. The frequency of these genotypes is various according to studies. Genotype 1 is predominant, except in Ghana where all the described isolates were genotype 3. A prospective French study performed between 1999 and 2001 indicated that genotypes 2 and 3 viruses circulated with a significant frequency (10%). Pathogenic properties might not differ according to the genotype. PMID- 18387750 TI - A unifying multiple sclerosis etiology linking virus infection, sunlight, and vitamin D, through viral interleukin-10. AB - Multiple sclerosis (MS) is a neurodegenerative disease of uncertain etiology. In MS, neurodegeneration is thought to be secondary to autoimmune-mediated damage. However, no cohesive explanation yet exists as to how environmental factors interact to induce a neurodegenerative autoimmune response. Insufficient sunlight exposure and chronic viral infections have been proposed as unrelated environmental risk factors for MS. We suggest that these risk factors may act synergistically to enable the pathogenic autoimmune response. Low ultraviolet light (UVL) exposure depletes vitamin D3 stores, and low vitamin D3 levels correlate strongly with high MS risk. The central nervous system converts vitamin D3 into 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D3), a biologically active hormone with anti-inflammatory and neuro-protective functions that depend on IL-10 producing regulatory lymphocytes. Herpesvirus infection also correlates with MS risk. Some herpesviruses like Epstein-Barr virus produce an IL-10-like cytokine termed vIL-10. We hypothesize that vIL-10 may induce a dysfunction of IL-10 producing regulatory lymphocytes, thereby undermining the protective functions of sunlight, vitamin D3, and 1,25-(OH)2 D3. The vIL-10 could elicit a host immune response capable of neutralizing or depleting IL-10, or the vIL-10 could compete with IL-10 but fail to perform an essential IL-10 function. In either case, the lack of sunlight exposure and the herpes virus infection might synergize to induce a defect in IL-10-producing regulatory lymphocyte function that undermines self-tolerance mechanisms and enables a pathogenic autoimmune response to neural proteins. PMID- 18387749 TI - Prepartying promotes heightened risk in the college environment: an event-level report. AB - Due to the emergence of research literature examining the prepartying behavior of college students, the present study examines students' varying drinking rates, blood alcohol levels (BALs), and alcohol-related consequences during two drinking events - one involving prepartying and one devoid of prepartying. Two-hundred and thirty-eight student drinkers completed an online drinking assessment detailing their two most recent drinking occasions involving and not involving prepartying. Participants responded to a series of questions regarding quantities consumed on the drinking day and occurrence of alcohol-related consequences. While men did not differ in drinking or estimated BALs, between the two drinking days, female participants drank significantly more drinks and reached higher BALs on the prepartying drinking day. Both males and females reported increased experience of alcohol-related consequences on the prepartying drinking day. In analyzing the prepartying drinking day specifically, we found that while men drink more alcohol during prepartying, but both men and women reached similar BALs during the event. Also, amount consumed during prepartying related to further drinking throughout the evening. It appears that prepartying may influence women to reach comparable levels of intoxication and alcohol-related consequences as their male peers. Quick drinking during prepartying may raise BALs and lead to alcohol-related consequences particularly for female students. PMID- 18387752 TI - [Clinical manifestations, complications and treatment of brucellosis: 45-patient study]. AB - The aim of this study was to evaluate the clinical, laboratory findings and therapeutic features of patients with brucellosis. The diagnosis was made by clinical findings, automated blood culture, serology (Rose Bengal plate agglutination test, standard tube agglutination (Wright) and immunofluorerescence). The susceptibility of 13 strains was tested in vitro. The base sequence was determined for four strains. Forty-five cases were collected (31 acute and 14 sub-acute). Contamination was digestive in 62%. Symptoms of patients were fever (93%), sweating (82%), arthralgia (78%) and splenomegaly (51%). Elevated erythrocyte sedimentation rate was determined in 80%, leukopenia in 49% and anaemia in 37% of cases. Blood cultures were positives in 39% of cases. The four sequenced strains were identified as Brucella melitensis biovar abortus. Six strains were resistant to sufomethoxazol-trimetoprim (54%). In 93% of cases, the treatment was associated rifampicin and doxycyclin. One patient died. No relapse was reported. PMID- 18387754 TI - Identification of conserved microRNAs and their target genes in tomato (Lycopersicon esculentum). AB - MicroRNAs (miRNAs) are a class of non-coding RNAs that have important gene regulation roles in various organisms. To date, a total of 1279 plant miRNAs have been deposited in the miRNA miRBase database (Release 10.1). Many of them are conserved during the evolution of land plants suggesting that the well-conserved miRNAs may also retain homologous target interactions. Recently, little is known about the experimental or computational identification of conserved miRNAs and their target genes in tomato. Here, using a computational homology search approach, 21 conserved miRNAs were detected in the Expressed Sequence Tags (EST) and Genomic Survey Sequence (GSS) databases. Following this, 57 potential target genes were predicted by searching the mRNA database. Most of the target mRNAs appeared to be involved in plant growth and development. Our findings verified that the well-conserved tomato miRNAs have retained homologous target interactions amongst divergent plant species. Some miRNAs express diverse combinations in different cell types and have been shown to regulate cell specific target genes coordinately. We believe that the targeting propensity for genes in different biological processes can be explained largely by their protein connectivity. PMID- 18387753 TI - Quantitative detection of bcr-abl transcripts in chronic myeloid leukemia. AB - The optimal management of malignant haematological disorders depend on the degree of tumor load reduction after therapy. Chronic myeloid leukemia constitutes a clinical model for molecular detection and therapy surveillance of malignant disease since this entity was the first leukemia shown to be associated with a specific bcr-abl fusion gene in the patient's leukemia cells. Molecular monitoring of bcr-abl transcript levels by real-time quantitative PCR is increasingly used to assess treatment response in patients with chronic myeloid leukemia (CML). This has become particularly relevant in the era of imatinib therapy when residual levels of leukaemia usually fall below the level of detection by bone marrow cytogenetic analysis. We monitored bcr-abl transcript levels by quantitative real time PCR in 50 tunisian patients treated with imatinib for chronic myeloid leukemia in chronic phase for a median of 29 months (3-60) after they started imatinib. PMID- 18387755 TI - Highly pathogenic avian influenza virus subtype H5N1 in ducks in the Northern part of Cameroon. AB - Highly pathogenic avian influenza (HPAI) virus was first detected in Cameroon in February 2006. Analysis of NA sequences of the virus demonstrated that it is closely related to the H5N1 isolates from Northern Nigeria, Sudan and Ivory Coast, suggesting a common virus ancestor. PMID- 18387756 TI - Tonsils--place of botulinum toxin production: results of routine laboratory diagnosis in farm animals. AB - Since a case of a veterinarian was reported, who was likely to be infected/intoxicated by Clostridium botulinum during the handling of a diseased animal, tonsils in animals were tested for botulinum neurotoxin and bacterial forms of neurotoxic Clostridium botulinum during routine botulism laboratory examinations including standard samples (intestinal tract and liver) from 48 cattle, 11 horses, and 14 goats. Ten out of 60 samples from tonsils contained free botulinum toxin, and 12 out of 59 were positive for live toxin producing bacteria. In 32 out of 162 intestinal samples toxin was detected. Toxin producing bacteria were found in 37 samples. Eight of 56 liver samples contained free toxin, and 15 out of 43 toxigenic bacteria. Samples from 10 slaughter pigs were all negative, whereas from slaughter cattle tonsils had a high incidence of toxin (7 of 10) or toxigenic bacteria (2 of 8). The results are discussed in the context of effects on animal health and botulism as zoonosis. PMID- 18387757 TI - Concomitant infection of cattle with the vaccine strain Anaplasma marginale ss centrale and field strains of A. marginale. AB - Bovine anaplasmosis, caused by Anaplasma marginale, the intraerythrocytic rickettsia, is controlled by vaccination with live Anaplasma marginale ss centrale (A. centrale), a subspecies of relatively low pathogenicity. We have experimentally demonstrated that an animal primarily infected with A. marginale, or with the related vaccine subspecies A. centrale can be infected with the heterologous subspecies, and carries both bacteria. The co-infection was detected in experimentally cross-infected calves for up to 3 months after the last inoculation with the heterologous subspecies. The occurrence of characteristic cyclic rickettsemia of A. centrale and A. marginale was observed by examination of Giemsa-stained blood smears, or by the presence of specific rickettsial DNA confirmed in PCR assays based on specific msp1a and msp4 for A. marginale, and on specifically designed msp3 and msp4 primers for A. centrale. Sequence analysis of msp4-specific fragments for each subspecies revealed the presence of dual infection in both calves on days 30 and 60 after cross-inoculation with the heterologous Anaplasma subspecies. The experimental cross-infection of calves clearly demonstrated that the concept of "infection exclusion" does not apply to Anaplasma infection in cattle; as there was no infection exclusion of A. marginale in A. centrale-infected cattle, and vice versa. The present results confirmed our previous findings that cattle grazing in an anaplasmosis-endemic field were subject to concomitant infection with both the vaccine A. centrale and the field A. marginale strains. PMID- 18387758 TI - Molecular detection and characterization of unclassified bovine enteric caliciviruses in South Korea. AB - The unclassified bovine enteric calicivirus (BEC) is a new bovine enteric calicivirus that is different from bovine norovirus, and causes diarrhea and pathologies in the small intestine of calves. This virus includes Nebraska (NB)- and Newbury agent 1 (NA1)-like strains. The prevalence of this BEC and its genetic characterization has only been reported in the UK and the USA. This study examined the prevalence and genetic diversity of these BECs in diarrheic calves in South Korea. Among a total of 645 diarrheic fecal specimens obtained from 629 cattle herds, these unclassified BECs were detected in 59 (9.1%) diarrheic fecal samples from 57 herds (9.3%) by either RT-PCR or nested PCR. Sequence and phylogenetic analyses of the partial RdRp gene showed that all the Korean BECs clustered together and were closely related to the NB-like viruses (80.9-88.1% nucleotide and 84.5-98.4% amino acid) but not to the NA1-like viruses (75.8-78.4% nucleotide and 79.7-82.8% amino acid). Although these viruses could not be classified into NA1- and NB-like viruses from the sequence and phylogenetic data of the entire capsid gene, all the Korean BECs clustered together on a branch separate from the other known BECs. These results show that these BEC infections are endemic in diarrheic calves in South Korea. The infecting strains are genetically closer to the NB-like viruses but have a distinct evolutionary pathway. PMID- 18387759 TI - Ultrasonographic-guided retrieval and in vitro maturation of eland (Taurotragus oryx) and bongo (Tragelaphus eurycerus isaaci) antelope oocytes. AB - The limited availability of gametes is a major factor hindering the development and application of assisted reproductive technologies (ART) in large non-domestic ungulates. This is partly due to the small number of captive animals and handling difficulties associated with procedures for gamete recovery. In the present study, results are reported of multi-year studies on ovarian stimulation and oocyte retrieval by ultrasonographic-guided transvaginal follicular aspiration and subsequent in vitro maturation (IVM) in eland and bongo antelopes. All procedures were conducted on sedated females handled in a hydraulic chute without inducing general anesthesia. Five estrous synchronization/ovarian stimulation protocols were evaluated and data are presented on 73 and 15 procedures in eland and bongo, respectively. Repeating procedures (< or =once/month) on the same female did not affect ovarian response or number oocytes recovered in either species. Eland females, but not the ovarian stimulation treatment, affected ovarian response. Ovarian stimulation treatment affected oocyte recovery rate in eland, but not in bongo. In both species, ovarian hormone stimulation treatment affected the distribution of follicles by size and the status of expansion of the cumulus cell investment of oocytes, but not the frequency of metaphase II oocytes during IVM. The timing of extrusion of the first polar body during IVM was more synchronous in bongo than in eland oocytes. It is concluded that Transvaginal oocyte retrieval (TVOR) can be safely and repeatedly applied in gonadotropin treated eland and bongo females to recover oocytes that can mature in vitro. The methods described for the present study can be adapted to improve the availability of non-domestic ungulate oocytes for basic and applied studies. PMID- 18387760 TI - Morphometric analysis of the urethra of castrated female rats treated with tamoxifen. AB - OBJECTIVES: The aim of this study was to evaluate the effects of tamoxifen on the weight and thickness of the urethral epithelium of castrated female rats. METHODS: Forty castrated adult female Wistar-Hannover rats were randomly divided into two groups: Group I (n=20) in which the animals received only the vehicle (propylene glycol) and Group II (n=20) in which the rats received tamoxifen 250microg/day by gavage. After 30 days of treatment, all animals were sacrificed and the urethra was immediately removed for weighing. Next, the urethra was divided into the proximal and distal segments, which were fixed in 10% formaldehyde and submitted to routine histological techniques for morphometric study. The data were analyzed using the weighted minimum mean-square error method and Student's t-test for two independent samples (p<0.05). RESULTS: There was a significant increase in the mean weight of the urethra in the rats of Group II compared to the control group, 32.0+/-2.0mg and 22.0+/-1.6mg, respectively (p<0.001). The mean thickness of the distal urethral epithelium of the animals treated with tamoxifen was significantly greater than that of the control group, 42.8+/-2.0microm and 36.6+/-1.5microm, respectively (p<0.001). There was no statistically significant difference between the two groups with respect to the epithelial thickness of the proximal urethra (p=0.514). CONCLUSION: Treating castrated adult rats with 250microg/day of tamoxifen for 30 days may increase the weight of the urethra and the thickness of the distal urethral epithelium. PMID- 18387762 TI - Where there is no PET/CT. AB - Technology assessment should touch upon four areas; utility, target population, cost and alternatives. Hybrid Positron Emission Tomography/Computed Tomography (PET/CT) is a promising modality for oncologic imaging. However, reservations about its accessibility and affordability prevail. Magnetic Resonance Imaging (MRI) is an alternative with certain key advantages that can potentially prove equivalent if not better than PET/CT. We have highlighted the value of comparative studies between PET/CT and whole body MRI. Diffusion Weighted sequences may aid in better diagnosis of malignancies on MRI. The 'Eye of Medicine' should not have a restricted vision even where there is no PET/CT. PMID- 18387763 TI - Carpal tunnel syndrome in Parkinson's disease. PMID- 18387764 TI - Sonography and electrodiagnosis in carpal tunnel syndrome diagnosis, an analysis of the literature. PMID- 18387761 TI - Effects of San-Huang-Xie-Xin-Tang on U46619-induced increase in pulmonary arterial blood pressure. AB - ETHNOPHARMACOLOGICAL RELEVANCE: San-Huang-Xie-Xin-Tang (SHXT), composed of Coptidis rhizoma, Scutellariae radix and Rhei rhizoma, is traditionally used to treat hypertension. AIM OF THE STUDY: Our aim was to investigate the pharmacology effect of SHXT on a thromboxane A(2) analogue U46619-induced increase in pulmonary hypertension and protein expression in primary pulmonary smooth muscle cells (PASMCs). MATERIALS AND METHODS: Arterial blood pressure and isometric tension in the aorta and pulmonary artery of rats were measured by pressure and force transducers, respectively. Protein expressions on PASMCs were detected by Western blotting. RESULTS: SHXT significantly attenuated U46619-induced increase in arterial blood pressure. The inhibitory effect of SHXT on pulmonary arterial pressure was greater than systemic arterial pressure in U46619 treated rats. Similarly, the inhibitory effect of SHXT on U46619-induced vasoconstriction in rat pulmonary arterial rings was greater than that in aortic rings. In U46619 treated PASMCs, SHXT down-regulated expression of phosphodiesterase type 5 (PDE5), Rho-kinase (ROCK) II, cyclooxygenase-2 (COX-2) and up-regulated expression of soluble guanylyl cyclase (sGC) alpha(1) and sGCbeta(1). CONCLUSIONS: SHXT attenuated U46619-induced increase in systemic and pulmonary arterial blood pressure. Inhibition of PDE5, ROCK-II, COX-2 and stimulation of sGC may play important roles in the cardiovascular effects of SHXT. PMID- 18387765 TI - Where there is no PET/CT. PMID- 18387766 TI - MRI of the rotator cuff and internal derangement. AB - Disease to the rotator cuff is the most common cause of shoulder pain and dysfunction in adults. This group of muscles performs multiple functions and is often stressed during various activities. The anatomy and physiology of the rotator cuff is complex and interconnected to other muscle groups in the shoulder. One must take the anatomic status of the rotator cuff tendons into account when planning the treatment of the rotator cuff injury. Diagnostic imaging of the rotator cuff, performed by MRI, provides valuable information about the nature of the injury. In this article, we will review the various types and causes of rotator cuff injuries, normal MR anatomy, function, patho-anatomy, and the biomechanics of the rotator cuff. We will also review shoulder impingement syndromes. PMID- 18387767 TI - Identification of major constituents in the traditional Chinese medicine "QI-SHEN YI-QI" dropping pill by high-performance liquid chromatography coupled with diode array detection-electrospray ionization tandem mass spectrometry. AB - An HPLC-DAD-ESI-MS(n) method was developed for simultaneous analysis of major chemical constituents in "QI-SHEN-YI-QI" dropping pill, a traditional Chinese medicine (TCM) widely used for treating cardiovascular diseases. The chromatographic separation was performed on an intertsil ODS-3 C(18) column (4.6 mm x 250 mm, 5 microm), whilst water with 0.05% acetic acid and acetonitrile were used as mobile phase. On the basis of the characteristic UV absorption profile, the information of molecular weight, and structure provided by ESI-MS(n), 31 constituents derived from Astragalus membranaceus, Radix Salviae Miltiorrhizae, and Panax notoginseng, were detected and 20 of them were identified in this study. The proposed method contributes to the quality control of "QI-SHEN-YI-QI" dropping pill. PMID- 18387768 TI - Liquid chromatography tandem mass spectrometry assay for the simultaneous determination of venlafaxine and O-desmethylvenlafaxine in human plasma and its application to a bioequivalence study. AB - A rapid, simple and sensitive liquid chromatography tandem mass spectrometry (LC MS/MS) assay for simultaneous determination of venlafaxine (VEN) and its active metabolite, O-desmethylvenlafaxine (ODV) in human plasma was developed using nadolol as internal standard (IS). The analytes and IS were extracted from 200 microl aliquots of human plasma via protein precipitation using 0.43% formic acid in acetonitrile and separated on a Hypurity cyano (50 mm x 4.6 mm, 5 microm) column. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique, operating in multiple reaction monitoring (MRM) and positive ion mode. The precursor to product ion transitions monitored for VEN, ODV and IS were m/z 278.3-->58.1, 264.3-->58.1 and 310.4- >254.1, respectively. The total chromatographic runtime was 3 min with retention time for VEN, ODV and IS at 1.93, 1.50 and 1.29 min, respectively. The method was fully validated for its sensitivity, accuracy and precision, linearity, recovery, matrix effect, dilution integrity and stability studies. The linear dynamic range of 2.0-500 ng/ml was established for both VEN and ODV with mean correlation coefficient (r), 0.9994 and 0.9990, respectively. The intra-batch and inter-batch precision (%CV) in three validation batches across five concentration levels (LLOQ, LQC, MQC, HQC and ULOQ) was less than 12.6% for both the analytes. The accuracy determined at these levels was within -9.8 to +3.9% in terms of %bias. The method was successfully applied to a bioequivalence study of 150 mg venlafaxine extended release capsule formulation in 22 healthy Indian male subjects under fed condition. PMID- 18387769 TI - Detection of Lipitor counterfeits: a comparison of NIR and Raman spectroscopy in combination with chemometrics. AB - Research has been carried on the feasibility of near infrared (NIR) and Raman spectroscopy as rapid screening methods to discriminate between genuine and counterfeits of the cholesterol-lowering medicine Lipitor. Classification, based on partial least squares discriminant analysis (PLS-DA) models, appears to be successful for both spectroscopic techniques, irrespective of whether atorvastatine or lovastatine has been used as the active pharmaceutical ingredient (API). The discriminative power of the NIR model, in particular, largely relies on the spectral differences of the tablet matrix. This is due to the relative large sample volume that is probed with NIR and the strong spectroscopic activity of the excipients. PLS-DA models based on NIR or Raman spectra can also be applied to distinguish between atorvastatine and lovastatine as the API used in the counterfeits tested in this study. A disadvantage of Raman microscopy for this type of analysis is that it is primarily a surface technique. As a consequence spectra of the coating and the tablet core might differ. Besides, spectra may change with the position of the laser in case the sample is inhomogeneous. However, the robustness of the PLS-DA models turned out to be sufficiently large to allow a reliable discrimination. Principal component analysis (PCA) of the spectra revealed that the conditions, at which tablets have been stored, affect the NIR data. This effect is attributed to the adsorption of water from the atmosphere after unpacking from the blister. It implies that storage conditions should be taken into account when the NIR technique is used for discriminating purposes. However, in this study both models based on NIR spectra and Raman data enabled reliable discrimination between genuine and counterfeited Lipitor tablets, regardless of their storage conditions. PMID- 18387770 TI - Microbial production of dihydroxyacetone. AB - Dihydroxyacetone is extensively used in cosmetic industry as an artificial suntan besides having clinical and biological applications. Thus, it is important to meet the commercial demand of dihydroxyacetone at an economical and qualitative level. Microbial route of production is found to be more favorable for dihydroxyacetone as compared to chemical methods. This review gives detailed information about the microbial route of dihydroxyacetone production. Till date the microorganism which is most utilized for dihydroxyacetone production is Gluconobacter oxydans. Some limitations associated with dihydroxyacetone production by G. oxydans like substrate inhibition, product inhibition and oxygen limitation are discussed here. Various fermentation modes and culture conditions have been tried for their ability to overcome these limitations. It has been found that fed-batch mode of fermentation provides a better yield as compared to batch mode for dihydroxyacetone production. Two-stage repeated fed-batch mode of fermentation has been found to be the most optimized mode. Immobilization has also been recognized as a much better alternative for fermentation since it avoids the problem of substrate and product inhibition to a greater extent. Although these methods have increased the dihydroxyacetone production to a prominent level yet the production has not reached the level required to meet the commercial demand. One looks for future prospects of developing recombinant microbial method for dihydoxyacetone production. PMID- 18387771 TI - Post-insult minocycline treatment attenuates hypoxia-ischemia-induced neuroinflammation and white matter injury in the neonatal rat: a comparison of two different dose regimens. AB - An increase in the number of activated microglia in the brain is a key feature of neuroinflammation after a hypoxic-ischemic insult to the preterm neonate and can contribute to white matter injury in the brain. Minocycline is a potent inhibitor of microglia and may have a role as a neuroprotective agent that ameliorates brain injury after hypoxia-ischemia in neonatal animal models. However to date large doses, pre-insult administration and short periods of treatment after hypoxia-ischemia have mostly been investigated in animal models making it difficult to translate minocycline's potential applicability to protect the human preterm neonatal brain exposed to hypoxia-ischemia. We investigated whether repeated doses of minocycline can minimize white matter injury and neuroinflammation one week after hypoxia-ischemia (right carotid artery ligation and 30 min 6% O(2)) in the post-natal day 3 rat pup. Two dosage regimens of minocycline were administered for one week; a high dose of 45 mg/kg 2h after hypoxia-ischemia then 22.5 mg/kg daily or a low dose 22.5 mg/kg 2h after hypoxia ischemia then 10 mg/kg. Post-natal day 3 hypoxia-ischemia significantly reduced myelin content, numbers of O1- and O4-positive oligodendrocyte progenitor cells and increased activated microglia one week later on post-natal day 10. The low dose minocycline regimen was as effective as the high dose in ameliorating neuroinflammation after post-natal day 3 hypoxia-ischemia. However only the high dose regimen significantly attenuated reductions in O1- and O4-positive oligodendrocyte progenitor cells and myelin content. The low dose only significantly attenuated the reduction in O1-positive oligodendrocyte cell counts. Repeated, daily, post-insult treatment with minocycline abolished neuroinflammation and may provide neuroprotection to white matter for up to one week after hypoxia-ischemia in a rodent preterm model. The present findings suggest the potential clinical relevance of a repeated, daily minocycline treatment strategy, administered after a hypoxia-ischemia insult, as a therapeutic intervention for hypoxia-ischemia-affected preterm neonates. PMID- 18387772 TI - International collaboration in shared decision-making: the International Shared Decision Making (ISDM) conference history and prospects. AB - OBJECTIVE: Analyze the role of collaboration in the International Shared Decision Making organization (ISDM). METHODS: Case study of the seven year history of ISDM as a professional network. RESULTS: The International Shared Decision Making meeting Conference (ISDM) has held four biennial meetings since its inception in 2001. It is a freestanding professional meeting, with no permanent institutional support and no formal governance structure. In both its history and its prospects, collaboration among attendees has been pivotal to its growth. It both attracts and holds its "members" through the strength of the relationships formed during and between meetings. Exchanges in ISDM are informational, collegial, and indirectly economic. CONCLUSION: ISDM's future rests on keeping all three functions healthy. It must maintain a focus on putting the shared decision making work first through becoming a "worknet". Technologies that promote shared decision making can then be assembled and developed that support transformation of health care. PRACTICE IMPLICATIONS: Innovative professional organizations need to develop their work through deliberate development of networking techniques to move innovation into practice. PMID- 18387773 TI - Meeting the health literacy needs of immigrant populations. AB - OBJECTIVE: Immigrant populations are vulnerable to serious health disparities, with many immigrants experiencing significantly worse health outcomes, such as higher rates of morbidity and mortality, than other segments of society. Immigrants disproportionately suffer from heart attacks, cancer, diabetes, strokes, HIV/AIDS, and many other serious diseases. These health risks demand effective health communication to help immigrants recognize, minimize, and respond effectively to potential health problems. Yet, while the need for effective communication about health risks is particularly acute, it is also tremendously complicated to communicate effectively with these vulnerable populations. METHODS: A literature review using online databases was performed. RESULTS: Immigrants often have significant language and health literacy difficulties, which are further exacerbated by cultural barriers and economic challenges to accessing and making sense of relevant health information. CONCLUSION: This paper examined the challenges to communicating relevant information about health risks to vulnerable immigrant populations and suggested specific communication strategies for effectively reaching and influencing these groups of people to reduce health disparities and promote public health. PRACTICE IMPLICATIONS: Communication interventions to educate vulnerable populations need to be strategic and evidence-based. It is important for health educators to adopt culturally sensitive communication practices to reach and influence vulnerable populations. Community participative communication interventions are a valuable strategy for integrating consumers' perspectives into health education efforts and building community commitment to health communication interventions. PMID- 18387774 TI - Women's preferences for their gynecologist or obstetrician. AB - OBJECTIVE: The objective of this study was to evaluate women's preferences in selecting their gynecologist or obstetrician. METHODS: This cross-sectional study was performed at the Women's Health Center "Netka", Tel Aviv over a 1-month period in 2006 and included 287 participants. Women were asked to complete an anonymous questionnaire. RESULTS: Most women (60.3%) reported that the gender of their gynecologist or obstetrician was not an important consideration when choosing a gynecologist or an obstetrician. The major determinants in their choice of a gynecologist or an obstetrician included professionalism (98.9%), courtesy (96.6%) and board certification (92%). The rating of the two most important factors in their choice revealed the following order: professionalism (45.3%), courtesy (25.8%), board certification (10.8%), availability (10.1%), comprehension (6.5%) and communication (1.5%). CONCLUSION: Israeli women's model of choice of their gynecologist involved physicians' professionalism and courtesy while availability and physician's gender was significantly less important. PRACTICE IMPLICATIONS: The presented data may help health providers during patients' education and counseling along with facilitating better understanding of patients' needs. PMID- 18387776 TI - Sclerotherapy after embolization of draining vein: a safe treatment method for venous malformations. AB - BACKGROUND: Treatment of congenital venous malformations poses a major clinical challenge. Great successes have been achieved with ethanol sclerotherapy in most lesions; however, severe complications are more likely to occur when more ethanol is used. OBJECTIVES: This study evaluated the safety and efficacy of a new sclerotherapy treatment that uses absolute alcohol and bleomycin A5. We evaluated our experience to present a safe treatment method for venous malformations. METHODS: The study population comprised 201 patients. Before treatment, lesions were categorized according to magnetic resonance imaging features: 120 patients had limited venous malformations, and 81 had infiltrating venous malformations. Percutaneous sclerotherapy was performed by direct injection of absolute alcohol and bleomycin A5 under fluoroscopy. The mean dose of the two medicines was 3.4 mL and 3.7 mg, respectively. We investigated 592 sclerotherapy sessions. A retrospective study was performed of medical records and color photographs and magnetic resonance imaging before and after treatment The mean follow-up was 29 months (range, 12-65 months). RESULTS: After embolization of the drainage vein with ethanol, a subsequent sclerosis therapy with bleomycin A5 was judged beneficial in 196 of 201 patients. Among the 196 responders, 56 showed disappearance of their chief symptoms, 42 showed improvement to nearly normal, and 62 showed marked improvement. The best response to our treatment was among the patients with limited venous malformations, with 114 (95%) showing at least marked improvement. Complications were tissue necrosis in 6 sessions, peripheral nerve palsy in 5, pigmentation in 10, blistering in 5, and 87 sessions resulted in transient side effects related to bleomycin A5, such as fever and gastrointestinal irritation. All were self-limited. No major complications such as cardiopulmonary collapse or pulmonary fibrosis were observed. CONCLUSION: Percutaneous sclerotherapy of venous malformations using absolute ethanol and bleomycin A5 is safe and effective. The simplicity, speed, and safety of sclerotherapy, combined with the quality and stability of the outcome achieved with our novel procedure, may make the technique the choice for treatment of venous malformations. PMID- 18387775 TI - Renal perfusion with venous blood extends the permissible suprarenal clamp time in abdominal aortic surgery. AB - OBJECTIVE: Suprarenal clamping in abdominal aortic surgery is associated with a significant risk of postoperative renal failure, general morbidity, and mortality. Arterial access for temporary shunting in these patients is difficult, and arterial perfusion techniques are typically complex. This study evaluated if renal perfusion with venous blood using a minimal pump setup and intermediate level heparinization prevents a decline in postoperative renal function in patients requiring suprarenal clamping for aortic reconstructive surgery. METHODS: Renal perfusion was achieved using a roller pump supplied with venous blood from a central venous catheter under medium-level heparinization to feed two perfusion balloon catheters. The calculated glomerular filtration rate was observed for 10 postoperative days and compared with the rate in patients with suprarenal clamping without renal perfusion. RESULTS: From 2001 to 2007, 158 patients underwent surgical reconstruction involving the pararenal aorta through a midline abdominal incision. Renal perfusion was started in 2006 and was always attempted if suprarenal clamping was anticipated preoperatively. Twenty-six patients received renal perfusion, and 132 also requiring suprarenal clamping did not. Of the latter, 109 were included in the control group. Five patients died or= 2 cm had an improved RFS from 80% to 88%, p=0.02. CONCLUSION: The increased use of adjuvant therapy was associated with an improved outcome in node positive patients. For node negative patients with good prognostic features the evidence of benefit was marginal. PMID- 18387778 TI - [Right colectomy with regional anaesthesia and hypnosedation in ASA 3 patient]. AB - In high-risk anaesthetic patients, the choice of a preoperative anaesthesia remains a difficult challenge before performing a heavy surgery such as colon excision. Nowadays, hypnosedation may be considered as an additional anaesthetic technique given to be associated with local or regional anaesthesia, in order to permit more surgery possibilities in high risk patients. PMID- 18387779 TI - [Treatment of bipolar disorders]. AB - Bipolar disorder is a hereditary, chronic, recurrent, and underdiagnosed disease. It involves mood dysregulation, associated with multiple comorbidities and residual symptoms between episodes. Prognosis worsens with delay of diagnosis and treatment. Neurological studies are essential to the understanding of this disorder. Cerebral lesions and progressive cognitive disorders have been shown. A well-conducted treatment strategy is fundamental. PMID- 18387780 TI - Genes regulating the serotonin metabolic pathway in the brain stem and their role in the etiopathogenesis of the sudden infant death syndrome. AB - Genotypes and allelic frequencies of TPH2, 5-HTTLPR, the 5-HTT (SLC6A4) intron 2 variable-number tandem repeat (VNTR) region, and the MAOA VNTR region were determined in brain-stem samples of 20 "genuine" SIDS cases and compared with results obtained from 150 healthy controls. The SNP G1463A responsible for 80% functionality loss of TPH2 (tryptophan hydroxylase 2) was not detected, neither in SIDS infants nor in the controls. In contrast, a strict relation was found between the 5-HTTLPR genotype and its allelic frequencies with SIDS cases. The L/L genotype and the long allele (L) of the promoter region of the serotonin transporter were significantly associated (likelihood ratio (LR) test, p<0.001) with the syndrome (L/L, 60% SIDS vs 14% controls; L, 80% SIDS vs 42.6% controls). Polymorphisms of the intron 2 VNTR of the same gene showed a trend for significant differences between genotypes 10/10 and 12/12 (LR test, p=0.068), with the L-12 haplotype being almost twofold in SIDS (44.5%) with respect to controls (23.4%). Differences were even higher considering the genotype combination L/L-12/12 (20% SIDS vs 2.6%), and variations among categories were statistically highly significant (p<0.001). Although additional differences were observed in the frequency of the MAOA (monoamine oxidase A) VNTR genotype 3R/3R between SIDS and controls (respectively 15% vs 26%), the results were not supported by statistical significance. Molecular polymorphisms are discussed considering their functional role in regulating serotonin synthesis (TPH2), neuronal reuptake (5-HTTLPR and 5-HTT intron 2), and catabolism (MAOA) in the nervous system of Italian SIDS infants. Comparisons are made with previous data obtained in different ethnic groups. PMID- 18387781 TI - Validation of the Finnvasc score in infrainguinal percutaneous transluminal angioplasty for critical lower limb ischemia. AB - The aim of the present study was to validate the Finnvasc score for prediction of immediate outcome after infrainguinal percutaneous transluminal angioplasty (PTA) for critical lower limb ischemia (CLI). Our registry included prospective data on 512 patients who underwent isolated infrainguinal PTA revascularization procedures for CLI. The Finnvasc score herein evaluated was calculated by assigning one point each to diabetes, coronary artery disease, foot gangrene, and urgent operation. Early mortality and major limb amputation rates after PTA revascularization were 2.5% and 12.3%, respectively. Seventy-two patients (14.1%) died and/or had lower limb amputation. Diabetes (p = 0.001), foot gangrene (p = 0.047), urgent operation (p < 0.0001), and preoperative renal failure (p = 0.001) were independent predictors of postoperative mortality and/or major limb amputation. Finnvasc score was predictive of major lower limb amputation (p = 0.003), mortality (p < 0.0001), and mortality and/or major amputation (p < 0.0001) after PTA. Mortality, major lower limb amputation, and combined end point rates in patients with a Finnvasc score of 3-4 were 12.8%, 25.6%, and 35.9%, respectively. The Finnvasc score is a simple risk scoring method which can be useful to estimate the risk of immediate postprocedural mortality and/or major lower limb amputation also in patients undergoing infrainguinal PTA for CLI. PMID- 18387783 TI - Effects of weekly exposure to anatoxin-a and nicotine on operant performance of rats. AB - This study examined the effects of acute and weekly administration of anatoxin-a and nicotine on operant performance. Anatoxin-a is a potent nicotinic receptor agonist produced by cyanobacteria, which are found in fresh waters throughout the world. Anatoxin-a is a potential human health hazard and has been responsible for numerous deaths of wildlife, livestock and domestic animals. Remarkably little is known, however, about the effects of anatoxin-a on behavior. Nicotine, the psychomotor stimulant in tobacco, has many well-documented behavioral effects, which often diminish (i.e. tolerance develops) when it is given daily. Male Long Evans rats initially were trained to respond under a multiple variable-ratio 30 response variable-interval 60-s (mult VR-30 VI 60-s) schedule of food reinforcement. They were then divided into 12 groups of 8 that received four weekly subcutaneous injections of anatoxin-a (0.05-0.2 mg/kg), nicotine (0.125 1.8 mg/kg), or vehicle 5-min prior to testing. When initially administered, each compound decreased response rates and reinforcement rates in both components of the multiple schedule. Substantial tolerance developed to the disruptive effects of nicotine with weekly administration. Tolerance also developed to the effects of anatoxin-a, although to a lesser degree; the highest dose severely decreased performance with little evidence of recovery. In conjunction with prior findings, these results suggest the behavioral effects of anatoxin-a and nicotine are similar, but not identical, and that relatively infrequent (episodic) administration can produce tolerance. PMID- 18387784 TI - Generalized fuzzy clustering for segmentation of multi-spectral magnetic resonance images. AB - An integrated approach for multi-spectral segmentation of MR images is presented. This method is based on the fuzzy c-means (FCM) and includes bias field correction and contextual constraints over spatial intensity distribution and accounts for the non-spherical cluster's shape in the feature space. The bias field is modeled as a linear combination of smooth polynomial basis functions for fast computation in the clustering iterations. Regularization terms for the neighborhood continuity of intensity are added into the FCM cost functions. To reduce the computational complexity, the contextual regularizations are separated from the clustering iterations. Since the feature space is not isotropic, distance measure adopted in Gustafson-Kessel (G-K) algorithm is used instead of the Euclidean distance, to account for the non-spherical shape of the clusters in the feature space. These algorithms are quantitatively evaluated on MR brain images using the similarity measures. PMID- 18387785 TI - To (TGF)beta or not to (TGF)beta: fine-tuning of Smad signaling via post translational modifications. AB - Smad proteins are key signal transducers for the TGF-beta superfamily and are frequently inactivated in human cancers, yet the molecular basis of how their levels and activities are regulated remains unclear. Recent progress, discussed herein, illustrates the critical roles of Smad post-translational modifications in the cellular outcome to TGF-beta signaling. PMID- 18387786 TI - The Early Childhood Epilepsy Severity Scale (E-Chess). AB - PURPOSE: We have developed the Early Childhood Epilepsy Severity Scale (E-Chess) to quantify the severity of epilepsy in infants and young children with tuberous sclerosis as an aid to the evaluation of treatment efficacy and the investigation of the influence of epilepsy severity on development. METHODS: Twenty infants aged 11-36 months with a diagnosis of tuberous sclerosis participated in the study. From the literature, six potential measures of epilepsy severity were identified: time period over which seizures occurred; seizure frequency; number of seizure types; occurrence and duration of status epilepticus; number of anticonvulsant medications used; response to treatment. The variables were given a score, usually from 0 to 3, a higher score indicating greater severity. For each child, these variables were scored over consecutive 1 year time periods by three independent raters. We employed restricted and nonrestricted factor analytic models to identify the latent structure of the six items. RESULTS: The six severity items had a unidimensional structure. All severity indicators loaded highly on the latent epilepsy severity factor (>0.77), with the exception of the status indicator which had a poor loading (<0.40) and was excluded from further analyses. Goodness of fit indices were all well within the acceptable criteria for model fit. The E-Chess score at 12 months was significantly predictive of scores at 24 and 36 months. CONCLUSIONS: A single continuous latent variable accounts for the variation in five of the six epilepsy severity indicators under study. These form the Early Childhood Epilepsy Severity Scale. The predictive validity of the E-Chess was satisfactory. The E-Chess provides an epilepsy severity score that can be easily used to assess epilepsy severity in tuberous sclerosis and would merit evaluation in other early onset childhood epilepsies. PMID- 18387789 TI - Improved understanding of the effect of food on drug absorption and bioavailability for lipophilic compounds using an intestinal pig perfusion model. AB - The purpose of this study was to investigate the relative importance of mechanisms behind the effect of food on the intestinal absorption and bioavailability for low solubility compounds by applying a porcine single-pass perfusion model. Nanoparticle suspensions of the model compounds, danazol and cyclosporine were perfused through the jejunum in isotonic fluid alone (control) and isotonic fluid with a P-glycoprotein (P-gp) inhibitor (verapamil) or dietary and endogenous lipids added. The drugs were also administered as saturated solutions in the isotonic fluid containing lipids. Administration of cyclosporine together with verapamil increased the absorption compared to the control (1.6 times) suggesting an effect on jejunal permeability. However, addition of dietary lipids to the media led to a 50% reduction in the absorption of cyclosporine indicating lack of major effects by P-gp inhibition by lipids in vivo. The absorption of danazol was increased (2.6 times) when administered as a nanosuspension in lipid containing media compared to the control, but decreased (60%) when administered as a solution in the same media. This shows how important dissolution of the drug nanoparticles is in drug absorption. The difference in the effect of lipids in the absorption of cyclosporine and danazol when administered as nanosuspensions may be due to different distribution to the colloidal structures present in the media, thereby rendering the drugs' different diffusion rates in the perfused segment. In conclusion, solubilisation seems to be a more important factor than P-gp inhibition as an explanation for the food drug interaction observed for several low solubility drugs. In addition, the partition into different colloidal structures seems to play a major role in the dissolution and absorption of poorly soluble drugs. PMID- 18387787 TI - Indicators of developmental deviance in individuals at risk for schizophrenia. AB - Schizophrenia is generally conceptualized as a neurodevelopmental disorder. In order to examine psychometrically-identified individuals at risk for schizophrenia in terms of indicators of developmental deviance, we examined digit ratios, nailfold plexus visibility, and dermatoglyphic features in young adults with elevated scores on the Social Anhedonia Scale. These individuals were compared to an age-matched control group. The two groups did not differ in terms of their digit ratios, though across both groups, the males had significantly lower 2D:4D ratios than the females. The socially anhedonic group had a significantly higher prevalence of nailfold plexus visibility. Males reporting excessive social anhedonia exhibited significantly lower a-b ridge counts than controls, though the two groups did not differ in terms of finger ridge counts. Study findings indicate that relationships exist between some indicators of nonspecific developmental injury and negative schizotypy, especially in males. PMID- 18387790 TI - Combining fulvestrant (Faslodex) with continued oestrogen suppression in endocrine-sensitive advanced breast cancer: the SoFEA trial. PMID- 18387788 TI - The psychotomimetic states inventory (PSI): measuring psychotic-type experiences from ketamine and cannabis. AB - BACKGROUND: This study reports a new measure of psychotomimetic states in the context of cannabis and ketamine use. The Psychotomimetic States Inventory (PSI) has sub-scales of Delusory Thinking, Perceptual Distortions, Cognitive Disorganization, Anhedonia, Mania and Paranoia. METHODS: The PSI was administered in two independent group, repeated measures designs: an experimental study of ketamine and a naturalistic study of cannabis. RESULTS: Both cannabis and ketamine produced reliable increases in ratings of psychotomimetic state effects across several sub-scales. CONCLUSIONS: The PSI is a potentially useful measure of the nature and extent of the phenomenological effects of psychotropic drugs in schizophrenia-related research. PMID- 18387796 TI - F2-isoprostane, inflammation, cardiac function and oxygenation in the endotoxaemic pig. AB - Prostaglandins are profoundly involved in endotoxaemic shock. Twenty pigs were given endotoxin at various doses (0.063-16 microg kg(-1) h(-1)). Three non endotoxaemic pigs served as controls. Two eicosanoids were measured in plasma (8 iso-PGF(2alpha), a free radical-mediated lipid peroxidation product, and 15-keto dihydro-PGF(2alpha) a major metabolite of COX activity) and evaluated against the pathophysiological responses that occur during endotoxaemic shock. Endotoxin mediates an increase in both 8-iso-PGF(2alpha) and 15-keto-dihydro-PGF(2alpha). An increase in the endotoxin dose induced significant log-linear responses in 8 iso-PGF(2alpha) and 15-keto-dihydro-PGF(2alpha). Oxidative injury correlated to the TNF-alpha, IL-6, reductions in cardiac performance and to oxygen delivery and utilisation. COX-mediated inflammatory responses correlated to TNF-alpha, IL-6 and to reductions in arterial oxygen tension. Thus, oxidative injury and COX mediated inflammation play a central role in the manifestation of endotoxaemic shock. Furthermore, formation of these eicosanoids on endotoxin-mediated alterations in pulmonary hypertension, oxygen delivery and oxygen utilisation seems to be independent of the administered endotoxin dose. PMID- 18387792 TI - Treatment of oesophageal squamous cell carcinoma in a patient with Fanconi anaemia. PMID- 18387794 TI - Development of the movement of the epiglottis in infant and juvenile pigs. AB - Although backward folding of the epiglottis is one of the signal events of the mammalian adult swallow, the epiglottis does not fold during the infant swallow. How this functional change occurs is unknown, but we hypothesize that a change in swallow mechanism occurs with maturation, prior to weaning. Using videofluoroscopy, we found three characteristic patterns of swallowing movement at different ages in the pig: an infant swallow, a transitional swallow and a post-weaning (juvenile or adult) swallow. In animals of all ages, the dorsal region of the epiglottis and larynx was held in an intranarial position by a muscular sphincter formed by the palatopharyngeal arch. In the infant swallow, increasing pressure in the oropharynx forced a liquid bolus through the piriform recesses on either side of a relatively stationary epiglottis into the esophagus. As the infant matured, the palatopharyngeal arch and the soft palate elevated at the beginning of the swallow, so exposing a larger area of the epiglottis to bolus pressure. In transitional swallows, the epiglottis was tilted backward relatively slowly by a combination of bolus pressure and squeezing of the epiglottis by closure of the palatopharyngeal sphincter. The bolus, however, traveled alongside but never over the tip of the epiglottis. In the juvenile swallow, the bolus always passed over the tip of the epiglottis. The tilting of the epiglottis resulted from several factors, including the action of the palatopharyngeal sphincter, higher bolus pressure exerted on the epiglottis and the allometry of increased size. In both transitional and juvenile swallows, the subsequent relaxation of the palatopharyngeal sphincter released the epiglottis, which sprang back to its original intranarial position. PMID- 18387800 TI - Aerobic granulation under the combined hydraulic and loading selection pressures. AB - Two SBR reactors were set up to investigate the feasibility of aerobic granulation under the combined selection pressures of hydraulic shear force and substrate loading. Aerobic granulation was studied at superficial upflow air velocity of 3.2 and 2.4 cm/s under an organic loading rate (OLR) range of 6.0 15.0 kg COD/m3d. Good reactor performance and well granule characteristics were achieved in a wide OLR range from 6.0 high up to 15.0 kg COD/m3d at 3.2 cm/s. While under the velocity of 2.4 cm/s, stable operation was limited in the OLR range of 6.0-9.0 kg COD/m3d and failed to operate with granule deterioration under further higher OLRs. The optimal combination of hydrodynamic shear force and loading selection pressure was demonstrated to be an important factor that influence aerobic granulation and govern the granule characteristics and reactor performance. PMID- 18387797 TI - Effect of disease duration on dose-response of inhaled budesonide in asthma. AB - BACKGROUND: Inhaled corticosteroids (ICS) represent first-line treatment in persistent asthma with clinical studies showing benefits of initiating therapy early. Whether treatment should be started with a high or low dose remains controversial. We investigated the importance of disease duration on the response to the starting dose of the ICS, budesonide, in asthma patients not previously treated with ICS. METHODS: Forty patients with newly detected asthma (symptoms for <12 months) and 41 patients with established asthma (mean duration 5.2 years, range 2-11) were randomized (double-blind, parallel-group) to treatment with budesonide Turbuhaler 100 or 400microg twice daily or placebo for 12 weeks. RESULTS: For morning peak expiratory flow (mPEF), all four budesonide treatments resulted in statistically significant improvements from baseline and, after 12 weeks, the changes in all four groups were statistically significantly greater than placebo. In patients receiving early treatment, no significant differences were seen between budesonide doses. In patients with established symptoms, 800 mg/day [corrected] improved mPEF significantly more than 200 mg/day [corrected] The 200 mg/day [corrected] dose in the early treatment group improved mPEF significantly more than in the delayed treatment group. Changes in forced expiratory volume in 1s (FEV(1)), the concentration of inhaled histamine causing a 20% drop in FEV(1), and use of as-needed medication behaved in very similar ways to mPEF. Asthma symptoms were reduced in all budesonide groups without a difference between doses. CONCLUSION: In patients with newly detected asthma treated early the initial ICS dose is not important. In contrast, in patients with symptoms for a longer duration a high starting dose improves airway function and hyperresponsiveness significantly better than a low dose. PMID- 18387801 TI - Long-term productivity of lowland and upland switchgrass cytotypes as affected by cutting frequency. AB - A considerable number of studies has been conducted on switchgrass (Panicum virgatum L.) as a bioresource for energy over the last few years. Nonetheless, some important issues concerning the agro-technique are still open. This research examines the long-term total dry matter yield (TDM) and ash content of two lowland (L) and two upland (U) switchgrass cytotypes, as affected by one or two cut system, under southern EU climatic conditions (44 degrees 33' N). Overall, L produced higher TDM than U (on average 14.9 and 11.7 Mg ha(-1), respectively); two-cut system allowed to produce higher biomass yields (especially in U) than single harvest during the two first years, but it also drastically reduced plant vigour and productivity of all cytotypes in the following two years. Moreover, under two-cut system almost total seasonal biomass derived from the early harvest, while the second cut slightly contributed to the total seasonal biomass, nor it appeared to offset the additional harvest costs. Biomass quality was also significantly affected by cutting frequency, with two-cut system leading to a considerably higher ash content of biomass. Therefore, it is perceived that two cut system is not worthwhile with U and L cytotypes as bioresource for energy production under southern EU conditions. PMID- 18387802 TI - Biobleaching of wheat straw-rich soda pulp with alkalophilic laccase from gamma proteobacterium JB: optimization of process parameters using response surface methodology. AB - An alkalophilic laccase from gamma-proteobacterium JB was applied to wheat straw rich soda pulp to check its bleaching potential by using response surface methodology based on central composite design. The design was employed by selecting laccase units, ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) concentration and pH as model factors. The results of second order factorial design experiments showed that all three independent variables had significant effect on brightness and kappa number of laccase-treated pulp. Optimum conditions for biobleaching of pulp with laccase preparation (specific activity, 65 nkat mg(-1) protein) were 20 nkat g(-1) of pulp, 2mM ABTS and pH 8.0 which enhanced brightness by 5.89% and reduced kappa number by 21.1% within 4h of incubation at 55 degrees C, without further alkaline extraction of pulp. Tear index (8%) and burst index (18%) also improved for laccase-treated pulp as compared to control raw pulp. Treatment of chemically (CEH1H2) bleached pulp with laccase showed significant effect on release of chromophores, hydrophobic and reducing compounds. Laccase-prebleaching of raw pulp reduced the use of hypochlorite by 10% to achieve brightness of resultant hand sheets similar to the fully chemically bleached pulp. PMID- 18387793 TI - The glycemic index of bread and biscuits is markedly reduced by the addition of a proprietary fiber mixture to the ingredients. AB - BACKGROUND AND AIM: Low glycemic index (GI) foods are associated with improved prevention and control of metabolic and cardiovascular diseases (e.g. diabetes and myocardial infarction), even if the impact of their consumption within mixed meals is difficult to predict. Since the availability of wheat based products maintaining taste and texture of traditional ones, but with a low GI, is of relevant nutritional interest, the aim of this study was to assess the effects of a specific proprietary fiber mix, added to the wheat flour used for biscuits and bread preparation, on their GI. METHODS AND RESULTS: Fifteen healthy non-diabetic volunteers ate on different days a portion equivalent to 75 g of available carbohydrates, of fiber enriched bread, traditional bread, fiber enriched biscuits and traditional biscuits, or a solution of 75 g of glucose in water. The glycemic index of each product was calculated by relating the area under their glycemic curve to that of glucose. The areas under the glycemic curves of fiber enriched bread and biscuits were lower than those obtained with the equivalent control food. Consequently, a marked reduction of their GI of 21% and 41% for bread and biscuits, respectively, was observed. CONCLUSION: The fiber mix added to the flour used in the preparation of biscuits and bread markedly reduces their GI. A similar effect could be expected in other oven-baked foods produced using the same fiber supplementation. PMID- 18387803 TI - A description of the characteristics of patients with non-ST elevation acute coronary syndromes admitted to different settings in the 1990s. AB - BACKGROUND: Little is known about the characteristics of patients who were admitted to the coronary care unit (CCU) in the 1990s with non-ST elevation acute coronary syndromes (ACS) compared with those admitted to medical and cardiology wards in hospitals in the United Kingdom (UK). AIM: To understand if there were systematic differences in the characteristics of patients who were admitted to UK critical care units (CCU), intermediate care environments (cardiology wards) or generalist wards (acute medical ward) in an observational study carried out in the 1990s. METHODS: This paper is based on a secondary analysis of PRAIS (UK), a prospective, observational, multi-centred study which recruited 1038 patients with non-ST elevation ACS. This analysis compares the characteristics of 860 of these patients dependent on whether they were cared for in a coronary care unit, acute medical ward or cardiology ward. RESULTS: The results showed that patients admitted to CCU were more likely to be younger, male, have a history of diabetes or have ST depression on their ECG. There was very little difference in other risk factors or prior concomitant therapy. Interestingly there were no systematic differences in treatments or outcomes other than would be expected by chance, although there were trends to higher rates of MI and heart failure in the CCU group. CONCLUSION: Our analysis shows that the main drivers of admission to CCU in the 1990s were ST depression and MI-both indicators of high risk, but older age and female gender seemed to decrease the likelihood of admission to CCU. Criteria for admission to specific specialist and non-specialist care environments should be standardised and the use of risk scores could be an important way forward. PMID- 18387798 TI - Plant biotechnology--predictive, green and quantitative. PMID- 18387791 TI - Radiofrequency ablation for locally recurrent colon carcinoma. PMID- 18387805 TI - Thick tellurium electrodeposition on nickel-coated copper substrate for 124I production. AB - Tellurium electrodeposition on a nickel-coated copper substrate was investigated for production of iodine-124. The electrodeposition experiments were carried out by the alkali plating baths. The optimum conditions of the electrodeposition of tellurium were as follows: 6 g l(-1) tellurium, pH=10, DC current density of ca. 8.55 mA cm(-2) and room temperature. PMID- 18387795 TI - Vertical distribution and phylogenetic composition of bacteria in the Eastern Tropical North Pacific Ocean. AB - The vertical community structure of bacteria along a depth profile in the Eastern Tropical North Pacific Ocean (13 degrees N, 104 degrees W) was studied by flow cytometry measurement and 16S rRNA gene clone libraries analysis. Picoeukaryotes and Synechococcus peaked at 30 m and decreased sharply below 50 m, while Prochlorococcus peaked at both 30 and 100 m layers and disappeared below 200 m. Heterotrophic bacteria peaked above shallow thermocline and decreased along the depth profile. Sequences of total 322 clones from four clone libraries (10, 100, 1000, and 3000 m) clustered into nine major lineages. gamma-Proteobacteria dominated all the depths and occupied almost the whole bacterial community at the 3000 m. alpha-Proteobacteria was abundant throughout the water column except near the sea bottom, and delta-Proteobacteria peaked at the 1000 m depth. Cyanobacteria were primarily limited to the photic zone, and the genetic diversity of Prochlorococcus showed a good correlation with niche adaptation. The appearance of the Cytophaga-Flexibacter-Bacteroides (CFB) group did not show a clear relationship with depth. Actinobacteria were found both in the photic zone and in deep water. Planctomyetes, Acidobacteria, and Verrucomicrobia were present as minor groups and more dominant in the deeper layers of water. PMID- 18387807 TI - Observation of beta-ray spectra after penetrating absorbing materials. AB - beta-Ray spectra after penetrating absorbing materials of various thicknesses were observed by the use of a scintillation-type beta-ray spectrometer equipped with a flat NE-102 plastic scintillator of 5mm thickness for sources of (60)Co, (90)Sr-(90)Y, (137)Cs, (147)Pm and (204)Tl. Although the spectra changed rapidly with increasing absorber thickness, the average beta-ray energy was kept nearly constant for a wider range. These results are consistent in that the beta-ray absorption curve becomes quasi-linear in a semi-logarithmic plot. Spectra including scattered beta-rays from several materials placed behind the source were also measured for (137)Cs and (204)Tl. It may be concluded that mean energy measurements by the use of beta-ray spectrometer of this kind is useful for the identification of nuclides in radiation protection purposes even in worse source conditions. PMID- 18387808 TI - A performance comparison of nine selected liquid scintillation cocktails. AB - Sample load capacity, sample compatibility, influence of sample load on (3)H/(14)C counting efficiency, background count rate, figure of merit, quench resistance, and sample stability for nine selected, commercially available, liquid scintillation cocktails are presented in this study. The cocktails tested are Ecoscint A, Insta-Gel Plus, OptiPhase Hisafe 3, OptiPhase Trisafe, Ready Gel, SafeScint 1:1, Ultima Gold, Ultima Gold LLT, and Ultima Gold XR. For the data acquisition a Packard TriCarb Model 1900CA and a Quantulus 1220 liquid scintillation counter is used. All samples are prepared in either 20mL low potassium, borosilicate glass vials or 20mL high density, polyethylene vials. The aim of the full comparison study is to determine a single cocktail that best suits all measurement requirements of the liquid scintillation laboratory at SCK.CEN for the determination of low levels of radioactivity in biological and environmental samples. The results published in this report are intended to help other laboratories in selecting a cocktail fit for their purposes or to set up a similar study. PMID- 18387806 TI - Experimental measurements and Monte Carlo calculations of dosimetric parameters of the IRA1-103Pd brachytherapy source. AB - This work presents a brachytherapy source having (103)Pd adsorbed onto a cylindrical silver rod that has been developed by Agricultural, Medical and Industrial Research School for permanent implant applications. Dosimetric characteristics (dose-rate constant, radial dose function, anisotropy function and anisotropy factor) of this source were experimentally and theoretically determined in terms of the updated AAPM Task Group 43 (TG-43U1) recommendations. Measurements were performed using TLD-GR200A circular chip dosimeters using standard methods employing thermoluminescent dosimeters in a Perspex phantom. Precision machined bores in the phantom located dosimeters and source in a reproducible fixed geometry providing for transverse-axis and angular dose profiles over a range of distances from 0.5 to 5 cm. The Monte Carlo N-Particle (MCNP) code, version 4C was used to evaluate the dose-rate distributions around this model (103)Pd source in water and Perspex phantoms. The Monte Carlo calculated dose-rate constant of the IRA1-(103)Pd source in water was found equal to Lambda=0.669 cGy/h/U with approximate uncertainties of +/-0.1%. The anisotropy function, F(r, theta), and the radial dose function, g(L)(r), of the IRA1-(103)Pd source were also measured in Perspex phantom and calculated in both Perspex and liquid water phantom. PMID- 18387799 TI - Brick by brick: metabolism and tumor cell growth. AB - Tumor cells display increased metabolic autonomy in comparison to non-transformed cells, taking up nutrients and metabolizing them in pathways that support growth and proliferation. Classical work in tumor cell metabolism focused on bioenergetics, particularly enhanced glycolysis and suppressed oxidative phosphorylation (the 'Warburg effect'). But the biosynthetic activities required to create daughter cells are equally important for tumor growth, and recent studies are now bringing these pathways into focus. In this review, we discuss how tumor cells achieve high rates of nucleotide and fatty acid synthesis, how oncogenes and tumor suppressors influence these activities, and how glutamine metabolism enables macromolecular synthesis in proliferating cells. PMID- 18387804 TI - Design, synthesis and trypanocidal activity of lead compounds based on inhibitors of parasite glycolysis. AB - The glycolytic pathway has been considered a potential drug target against the parasitic protozoan species of Trypanosoma and Leishmania. We report the design and the synthesis of inhibitors targeted against Trypanosoma brucei phosphofructokinase (PFK) and Leishmania mexicana pyruvate kinase (PyK). Stepwise library synthesis and inhibitor design from a rational starting point identified furanose sugar amino amides as a novel class of inhibitors for both enzymes with IC(50) values of 23microM and 26microM against PFK and PyK, respectively. Trypanocidal activity also showed potency in the low micromolar range and confirms these inhibitors as promising candidates for the development towards the design of anti-trypanosomal drugs. PMID- 18387809 TI - Comparative study of two drying techniques used in radioactive source preparation: freeze-drying and evaporation using hot dry nitrogen jets. AB - Quantitative solid sources are used widely in the field of radionuclide metrology. With the aim to improve the detection efficiency for electrons and x rays, a comparative study between two source drying techniques has been undertaken at LNE-Laboratoire National Henri Becquerel (LNE-LNHB, France). In this paper, freeze-drying using commercial equipment is compared with a system of drying using hot jets of nitrogen developed at Institute for Reference Materials and Measurements (IRMM, Belgium). In order to characterize the influence of self absorption, the detection efficiencies for (51)Cr sources have been measured by coincidence counting and photon spectrometry. PMID- 18387810 TI - A novel technique for the effective concentration of 99mTc from a large alumina column loaded with low specific-activity (n,gamma)-produced 99Mo. AB - Molybdenum-99 of low-to-medium specific activity, 7.4-14.8 GBq/g, obtained by the (98)Mo(n,gamma)(99)Mo reaction in research reactors requires a large alumina column to adsorb the molybdenum since the capacity of alumina to adsorb Mo is limited (20 mg Mo/g of alumina). The large size of alumina column, in turn, requires large eluant volumes to elute the (99m)Tc, and the radioactive concentration (RAC) of the pertechnetate becomes unacceptably low for radiopharmaceutical applications. This study describes a method for obtaining high RAC pertechnetate solutions from a column generator loaded with (99)Mo of low specific activity. Molybdenum-99 of specific activity, 7.4 GBq/g, was absorbed on an alumina column [60 g, 70 mm, (H) x 35 mm (diam.)]. The (99m)TcO(4)(-), eluted with 60 ml normal saline (0.15N NaCl), was applied to a 12 mm (H) x 1 mm (diam.) column containing 15 mg of the strong anion exchanger Dowex 1 x 8. The retained pertechnetate was eluted with 5 ml of 0.5 mM tetrabutylammonium bromide (TBAB) solution in methylene chloride. The organic solvent was subsequently removed under vacuum and the dry residue was reconstituted with the desired volume of normal saline. All the operations were carried out in a closed cycle with sterile connecting tubes and multi-way stop cocks. The concentrated pertechnetate obtained contained about 90% of the total saline eluate and had a pH of 6-7, a radiochemical purity (RCP) consistently >99%, the (99)Mo breakthrough was around 10(-4)%. Colorimetric spot tests (limit tests) for Al and Mo showed <10 ppm in the final (99m)TcO(4)(-) solution. The RCP of the labelled compounds prepared using the pertechnetate obtained and five different radiopharmaceutical kits were >95%. The procedure may be equally applicable for the effective concentration of (188)ReO(4)(-) from (188)W-(188)Re generators. PMID- 18387811 TI - Homer2 and Homer3 interact with amyloid precursor protein and inhibit Abeta production. AB - The study of Amyloid Precursor Protein (APP) processing has been the focus of considerable interest, since it leads to Abeta peptide generation, the main constituent of neuritic plaques found in brains of Alzheimer's disease patients. Therefore, the identification of novel APP binding partners that regulate Abeta peptide production represents a pharmaceutical target aiming at reducing Alphabeta pathology. In this study, we provide evidence that Homer2 and Homer3 but not Homer1 proteins interact specifically with APP. Their expression inhibits APP processing and reduces secretion of Abeta peptides. In addition, they decrease the levels of cell surface APP and inhibit maturation of APP and beta secretase (BACE1). The effects of Homer2 and Homer3 on APP trafficking to the cell surface and/or on APP and BACE1 maturation could be part of the mechanism by which the expression of these proteins leads to the significant reduction of Abeta peptide production. PMID- 18387814 TI - Traumatic brain injury: an integrated clinical case presentation and literature review part II: the continuum of care. AB - The following paper continues the presentation of a case scenario outlining the assessment, interventions and outcome of a person who sustained multiple trauma with a focus on traumatic brain injury (TBI). Part I explored assessment and initial management of the patient from pre-hospital care through to the emergency department and operating theatre. Part II describes the intensive care period as an integral component of the continuum of care. Key issues in the case are presented sequentially with relevant theory integrated and applied to the clinical case throughout the discussion with a focus on the complex physiological, psychological, and spiritual needs of the patient and their family. PMID- 18387815 TI - Traumatic brain injury: an integrated clinical case presentation and literature review. Part I: assessment and initial management. AB - Holistic nursing care of critically ill patients continues to be a challenge for all levels of critical care clinicians. Patients with multi-system dysfunction in particular, present complicated clinical challenges that demand care based on sound knowledge and understanding of physiological, psychosocial and spiritual needs. Experiential learning through exposure to a range of patient presentations enables incremental development of professional practice and excellence in nursing care. Case study learning enhances understanding through application of theory to practice in complex clinical presentations. This two-part paper outlines the assessment, interventions and outcome of a person who sustained multiple trauma including severe traumatic brain injury (TBI). Part I explores assessment and initial management from pre-hospital care through to the Emergency Department (ED) and operating theatre. Part II describes the intensive care period as an integral component of the continuum of care. Key issues in the case are presented sequentially with relevant literature integrated and applied to clinical progress, focussing on the complex physiological, psychosocial, spiritual and environmental needs of the patient and his family. The purpose of the paper is to therefore provide a comprehensive learning resource for critical care nurses, particularly for those beginning their practice. PMID- 18387813 TI - Errors in preparation and administration of intravenous medications in the intensive care unit of a teaching hospital: an observational study. AB - OBJECTIVE: To determine the frequency of medication errors that occurred during the preparation and administration of IV drugs in an intensive care unit. SETTING: The study was conducted in a 12-bed intensive care unit of one of the largest teaching hospitals in Tehran. DESIGN: Data were collected over 16 randomly selected days at different medication round times, between July and September 2006. A trained observer accompanied nurses during intravenous (IV) drug rounds. Medication errors were recorded during the observation times of IV drug administration and preparation. Drugs with the highest rate of use in the intensive care unit (ICU) were selected. Details of the process of preparation and administration of the selected drugs were compared to an informed checklist which was prepared using reference books and manufacturers' instructions. RESULTS: We observed a total of 524 preparations and administrations. The calculated number of opportunities for error was 4040. The number of errors identified were 380/4040 (9.4%). Of those, 33.6% were related to the preparation process and 66.4% to the administration process. The most common type of error (43.4%) was the injection of bolus doses faster than the recommended rate. Amikacin was involved in the highest rate of error (11%) among all the selected medications. It was found that the IV rounds conducted at 9:a.m. had the highest rate of error (19.8%). No significant correlation was found between the rate of error and the nurses' age, sex, qualification, work experience, marital status, and type of working contract (permanent or temporary). CONCLUSIONS: Since our system is devoid of a well-organized reporting system, errors are not detected and consequently not prevented. Administrators need to take the initiative of developing systems that guarantee safe medication administration. PMID- 18387812 TI - A seizure-induced gain-of-function in BK channels is associated with elevated firing activity in neocortical pyramidal neurons. AB - A heritable gain-of-function in BK channel activity has been associated with spontaneous seizures in both rodents and humans. We find that chemoconvulsant induced seizures induce a gain-of-function in BK channel current that is associated with abnormal, elevated network excitability. Action potential half width, evoked firing rate, and spontaneous network activity in vitro were all altered 24 h following picrotoxin-induced seizures in layer 2/3 pyramidal cells in the neocortex of young mice (P13-P16). Action potential half-width and firing output could be normalized to control values by application of BK channel antagonists in vitro. Thus, both inherited and acquired BK channel gain-of functions are linked to abnormal excitability. Because BK channel antagonists can reduce elevated firing activity in neocortical neurons, BK channels might serve as a new target for anticonvulsant therapy. PMID- 18387818 TI - Neuroendocrine cultured cells counteract persistent prion infection by down regulation of PrPc. AB - Cell models for prion diseases are mainly of neuronal origin. However, the pathological isoform PrP(Sc) of cellular prion protein (PrP(c)) and prion infectivity are found in a variety of extraneural tissues in prion diseases. Although many cell types are not able to propagate PrP(Sc), little is known about cellular mechanism counteracting prion infection. It is desirable to identify neuronal or non-neuronal cell models that restrict PrP(Sc) generation or propagate PrP(Sc) only transiently. Neuroendocrine cells are derived from tumours forming the interface between endocrine and nervous system. We investigated the susceptibility of such murine cell lines to prion infection, which were in principle able to transiently propagate PrP(Sc). Surprisingly and in contrast to neuronal cells prion infection was abrogated by rapid and PrP(Sc)-specific down regulation of PrP(c) expression upon exposure to prion-infected material. Cell lines described here provide novel models for studying PrP(c) regulation and intrinsic cellular defence mechanisms upon prion exposure. PMID- 18387819 TI - Cloning, expression, purification and characterization of a DsbA-like protein from Wolbachia pipientis. AB - Wolbachia pipientis are obligate endosymbionts that infect a wide range of insect and other arthropod species. They act as reproductive parasites by manipulating the host reproduction machinery to enhance their own transmission. This unusual phenotype is thought to be a consequence of the actions of secreted Wolbachia proteins that are likely to contain disulfide bonds to stabilize the protein structure. In bacteria, the introduction or isomerization of disulfide bonds in proteins is catalyzed by Dsb proteins. The Wolbachia genome encodes two proteins, alpha-DsbA1 and alpha-DsbA2, that might catalyze these steps. In this work we focussed on the 234 residue protein alpha-DsbA1; the gene was cloned and expressed in Escherichia coli, the protein was purified and its identity confirmed by mass spectrometry. The sequence identity of alpha-DsbA1 for both dithiol oxidants (E. coli DsbA, 12%) and disulfide isomerases (E. coli DsbC, 14%) is similar. We therefore sought to establish whether alpha-DsbA1 is an oxidant or an isomerase based on functional activity. The purified alpha-DsbA1 was active in an oxidoreductase assay but had little isomerase activity, indicating that alpha DsbA1 is DsbA-like rather than DsbC-like. This work represents the first successful example of the characterization of a recombinant Wolbachia protein. Purified alpha-DsbA1 will now be used in further functional studies to identify protein substrates that could help explain the molecular basis for the unusual Wolbachia phenotypes, and in structural studies to explore its relationship to other disulfide oxidoreductase proteins. PMID- 18387817 TI - Antiangiogenic effects of flavonoids and chalcones. AB - Angiogenesis, the development of new blood vessels from the existing vasculature, is essential in normal developmental processes. Uncontrolled angiogenesis is a major contributor to a number of disease states such as inflammatory disorders, obesity, asthma, diabetes, cirrhosis, multiple sclerosis, endometriosis, AIDS, bacterial infections and autoimmune disease. It is also considered a key step in tumour growth, invasion, and metastasis. Angiogenesis is required for proper nourishment and removal of metabolic wastes from tumour sites. Therefore, modulation of angiogenesis is considered as therapeutic strategies of great importance for human health. Numerous bioactive plant compounds are recently tested for their antiangiogenic potential. Among the most frequently studied are polyphenols present in fruits and vegetables. Plant polyphenols inhibit angiogenesis and metastasis through regulation of multiple signalling pathways. Specifically, flavonoids and chalcones regulate expression of VEGF, matrix metalloproteinases (MMPs), EGFR and inhibit NFkappaB, PI3-K/Akt, ERK1/2 signalling pathways, thereby causing strong antiangiogenic effects. This review focuses on the antiangiogenic properties of flavonoids and chalcones and examines underlying mechanisms. PMID- 18387820 TI - The enigma of the haemogram "left-shift" in periwinkles infected with trematodes. AB - To make further progress in the understanding of digenean immune evasive tactics in their snail host, we compared the haemopoietic parameters and haemocyte functional potencies in the prosobranch Littorina littorea, which were either healthy or infected with echinostome trematode Himasthla elongata. The haemocyte concentration in the circulation of infected individuals was significantly higher than in uninfected ones, 3300 microl(-1) and 1882 microl(-1), respectively. Intense haemopoiesis in haemolymph of infected snails was evidenced by 4-fold higher BrdU incorporation into nuclei of haemocytes as well as elevated level of cyclin D expression in these cells. Evident skewing of the haemocyte population toward a higher frequency of immature, undifferentiated haemocytes in infected L. littorea was found. Haemocytes in infected snails had a much lower functional potency in production of reactive oxygen intermediates (ROI) and cell-mediated cytotoxicity. Correlation analysis shows that both cytotoxic and ROI generation values were significantly and negatively correlated with proportion of juvenile cells in circulation. Experimental injection of H. elongata excretory/secretory products modulated haemopoiesis toward increasing a juvenile cells proportion by 7th day post-injection. This can be considered a haemogram "left-shift" by analogy to that seen in the human neutrophil compartment, when more immature bandforms are found in blood during acute inflammation or bone marrow disorders. We hypothesize that echinostomatide trematodes may interfere in the normal neuroendocrine management of haemopoiesis in the host and cause a haemopoietic signal to initiate multiplication to near neoplastic levels. PMID- 18387816 TI - Peptide-targeted radionuclide therapy for melanoma. AB - Melanocortin-1 receptor (MC1-R) and melanin are two attractive melanoma-specific targets for peptide-targeted radionuclide therapy for melanoma. Radiolabeled peptides targeting MC1-R/melanin can selectively and specifically target cytotoxic radiation generated from therapeutic radionuclides to melanoma cells for cell killing, while sparing the normal tissues and organs. This review highlights the recent advances of peptide-targeted radionuclide therapy of melanoma targeting MC1-R and melanin. The promising therapeutic efficacies of 188Re-(Arg(11))CCMSH (188Re-[Cys(3,4,10), D-Phe(7),Arg(11)]-alpha-MSH(3-13)), 177Lu- and 212Pb-labeled DOTA-Re(Arg(11))CCMSH (1,4,7,10-tetraazacyclododecane 1,4,7,10-tetraacetic acid-[ReO-(Cys(3,4,10), D-Phe(7), Arg(11))]-alpha-MSH(3-13)) and 188Re-HYNIC-4B4 (188Re-hydrazinonicotinamide-Tyr-Glu-Arg-Lys-Phe-Trp-His-Gly Arg-His) in preclinical melanoma-bearing models demonstrate an optimistic outlook for peptide-targeted radionuclide therapy for melanoma. Peptide-targeted radionuclide therapy for melanoma will likely contribute in an adjuvant setting, once the primary tumor has been surgically removed, to treat metastatic deposits and for treatment of end-stage disease. The lack of effective treatments for metastatic melanoma and end-stage disease underscores the necessity to develop and implement new treatment strategies, such as peptide-targeted radionuclide therapy. PMID- 18387821 TI - Diffusion-based spatial priors for functional magnetic resonance images. AB - We recently outlined a Bayesian scheme for analyzing fMRI data using diffusion based spatial priors [Harrison, L.M., Penny, W., Ashburner, J., Trujillo-Barreto, N., Friston, K.J., 2007. Diffusion-based spatial priors for imaging. NeuroImage 38, 677-695]. The current paper continues this theme, applying it to a single subject functional magnetic resonance imaging (fMRI) study of the auditory system. We show that spatial priors on functional activations, based on diffusion, can be formulated in terms of the eigenmodes of a graph Laplacian. This allows one to discard eigenmodes with small eigenvalues, to provide a computationally efficient scheme. Furthermore, this formulation shows that diffusion-based priors are a generalization of conventional Laplacian priors [Penny, W.D., Trujillo-Barreto, N.J., Friston, K.J., 2005. Bayesian fMRI time series analysis with spatial priors. NeuroImage 24, 350-362]. Finally, we show how diffusion-based priors are a special case of Gaussian process models that can be inverted using classical covariance component estimation techniques like restricted maximum likelihood [Patterson, H.D., Thompson, R., 1974. Maximum likelihood estimation of components of variance. Paper presented at: 8th International Biometrics Conference (Constanta, Romania)]. The convention in SPM is to smooth data with a fixed isotropic Gaussian kernel before inverting a mass univariate statistical model. This entails the strong assumption that data are generated smoothly throughout the brain. However, there is no way to determine if this assumption is supported by the data, because data are smoothed before statistical modeling. In contrast, if a spatial prior is used, smoothness is estimated given non-smoothed data. Explicit spatial priors enable formal model comparison of different prior assumptions, e.g., that data are generated from a stationary (i.e., fixed throughout the brain) or non-stationary spatial process. Indeed, for the auditory data we provide strong evidence for a non-stationary process, which concurs with a qualitative comparison of predicted activations at the boundary of functionally selective regions. PMID- 18387822 TI - Optimization of diffusion spectrum imaging and q-ball imaging on clinical MRI system. AB - Mapping complex crossing fibers using diffusion MRI techniques requires adequate angular precision and accuracy. Beyond diffusion tensor imaging (DTI), high angular resolution sampling schemes such as diffusion spectrum imaging (DSI) and q-ball imaging (QBI) were proposed to resolve crossing fibers. These schemes require hundreds of data approximately five to ten times more than DTI, offsetting their clinical feasibility. To facilitate its clinical application, optimum values of highest diffusion sensitivity (bmax) must be investigated under the constraint of scan time and gradient performance. In this study, simulation of human data sets and a following verification experiment were performed to investigate the optimum bmax of DSI and QBI. Four sampling schemes, two with high sampling number, i.e., DSI515 and QBI493, and two with low sampling number, i.e., DSI203 and QBI253, were compared. Deviation angle and angular dispersion were used to evaluate the precision and accuracy among different bmax of each scheme. The results indicated that the optimum bmax was a trade-off between SNR and angular resolution. At their own optimum bmax, the reduced sampling schemes yielded angular precision and accuracy comparable to the high sampling schemes. On our current 3 T system, the optimum bmax (s/mm2) were 6500 for DSI515, 4000 for DSI203, 3000 for QBI493 and 2500 for QBI253. DSI was incrementally more accurate than QBI, but required a greater demand for gradient performance. In conclusion, our systematic study of optimum bmax in different sampling schemes and the consideration derived wherein could be helpful to determine optimum sampling schemes in other MRI systems. PMID- 18387824 TI - Connectivity modulation of early visual processing areas during covert and overt tracking tasks. AB - The brain regions for pursuit and saccadic eye movement processing are well known. There is, however, little knowledge about the interaction between these areas during voluntary eye movements. With 8 subjects, we investigated the dynamics of cortical areas involved in control of saccadic and smooth pursuit eye movements. We explored the connectivity between V1, hMT+, and LIP. Additionally, we explored the effects caused by shifting covert attention between pursuit and saccade targets. We modeled 15 plausible models, selecting the best one using a new group comparison approach for DCM models. Effective connectivity from V1 to hMT+ was shown to depend on whether subjects attended covertly or overtly to the targets. Comparing active tracking tasks resulted in effects in accordance with current theories of the eye movement processing system. PMID- 18387826 TI - Cortical thickness measured from MRI in the YAC128 mouse model of Huntington's disease. AB - A recent study found differences in localised regions of the cortex between the YAC128 mouse model of Huntington's Disease (HD) and wild-type mice. There are, however, few tools to automatically examine shape differences in the cortices of mice. This paper describes an algorithm for automatically measuring cortical thickness across the entire cortex from MRI of fixed mouse brain specimens. An analysis of the variance of the method showed that, on average, a 50 microm (0.05 mm) localised difference in cortical thickness can be measured using MR scans. Applying these methods to 8-month-old YAC128 mouse model mice representing an early stage of HD, we found an increase in cortical thickness in the sensorimotor cortex, and also revealed regions wherein decreasing striatal volume correlated with increasing cortical thickness, indicating a potential compensatory response. PMID- 18387823 TI - Modeling motor connectivity using TMS/PET and structural equation modeling. AB - Structural equation modeling (SEM) was applied to positron emission tomographic (PET) images acquired during transcranial magnetic stimulation (TMS) of the primary motor cortex (M1(hand)). TMS was applied across a range of intensities, and responses both at the stimulation site and remotely connected brain regions covaried with stimulus intensity. Regions of interest (ROIs) were identified through an activation likelihood estimation (ALE) meta-analysis of TMS studies. That these ROIs represented the network engaged by motor planning and execution was confirmed by an ALE meta-analysis of finger movement studies. Rather than postulate connections in the form of an a priori model (confirmatory approach), effective connectivity models were developed using a model-generating strategy based on improving tentatively specified models. This strategy exploited the experimentally imposed causal relations: (1) that response variations were caused by stimulation variations, (2) that stimulation was unidirectionally applied to the M1(hand) region, and (3) that remote effects must be caused, either directly or indirectly, by the M1(hand) excitation. The path model thus derived exhibited an exceptional level of goodness (chi(2)=22.150, df=38, P=0.981, TLI=1.0). The regions and connections derived were in good agreement with the known anatomy of the human and primate motor system. The model-generating SEM strategy thus proved highly effective and successfully identified a complex set of causal relationships of motor connectivity. PMID- 18387825 TI - High spatial resolution increases the specificity of block-design BOLD fMRI studies of overt vowel production. AB - Functional MRI (fMRI) studies of tasks involving orofacial motion, such as speech, are prone to problems related to motion-induced magnetic field variations. Orofacial motion perturbs the static magnetic field, leading to signal changes that correlate with the task and corrupt activation maps with false positives or signal loss. These motion-induced signal changes represent a contraindication for the implementation of fMRI to study the neurophysiology of orofacial motion. An fMRI experiment of a structured, non-semantic vowel production task was performed using four different voxel volumes and three different slice orientations in an attempt to find a set of acquisition parameters leading to activation maps with maximum specificity. Results indicate that the use of small voxel volumes (2 x 2 x 3 mm(3)) yielded a significantly higher percentage of true positive activation compared to the use of larger voxel volumes. Slice orientation did not have as great an impact as spatial resolution, although coronal slices appeared superior at high spatial resolutions. Furthermore, it was found that combining the strategy of high spatial resolution with an optimum task duration and post-processing methods for separating true and false positives greatly improved the specificity of single-subject, block-design fMRI studies of structured, overt vowel production. PMID- 18387827 TI - Abnormal P300 in people with high risk of developing psychosis. AB - BACKGROUND: Individuals with an "at-risk mental state" (or "prodromal" symptoms) have a 20-40% chance of developing psychosis; however it is difficult to predict which of them will become ill on the basis of their clinical symptoms alone. We examined whether neurophysiological markers could help to identify those who are particularly vulnerable. METHOD: 35 cases meeting PACE criteria for the at-risk mental state (ARMS) and 57 controls performed an auditory oddball task whilst their electroencephalogram was recorded. The latency and amplitude of the P300 and N100 waves were compared between groups using linear regression. RESULTS: The P300 amplitude was significantly reduced in the ARMS group [8.6+/-6.4 microvolt] compared to controls [12.7+/-5.8 microvolt] (p<0.01). There were no group differences in P300 latency or in the amplitude and latency of the N100. Of the at-risk subjects that were followed up, seven (21%) developed psychosis. CONCLUSION: Reduction in the amplitude of the P300 is associated with an increased vulnerability to psychosis. Neurophysiological and other biological markers may be of use to predict clinical outcomes in populations at high risk. PMID- 18387829 TI - Heat shock protein 72 associated with CD44v6 in human colonic adenocarcinoma. AB - CD44v6 is a splice variant of CD44 (CD44v), probably promoting cancer cell adherence to vascular endothelium and base membranes and enhancing the invasion and metastasis of colonic carcinomas. Heat shock protein 72 (HSP72) as a molecular chaperone has been confirmed to be overexpressed in epithelial carcinoma cells. There may be a possible association between the expression of HSP72 and CD44v6 during the growth and progression of colonic carcinoma cells. The aim of the study was to investigate the interaction between heat shock protein 72 and CD44v6 in human colonic carcinomas. The localization of HSP72 and CD44v6 in human colonic carcinomas was determined by immunohistochemistry and confocal laser microscopy. The interaction between HSP72 and CD44v6 in colonic carcinoma cells was analyzed by immunoprecipitation and Western immunoblots. Our results revealed that colonic carcinoma synchronously co-expressed higher levels of HSP72 and CD44v6 than that in adjacent normal colonic tissues. HSP72 and CD44v6 were mainly immunolocalized in the cytoplasm, and also immunolabelled on the cell membrane. Based on immunoprecipitation and Western immunoblots, we found that HSP72 was associated with CD44v6 precursor fragments in human colonic carcinoma cells. The interaction between HSP72 and CD44v6 in human colonic carcinoma cells may contribute to study the pathogenesis and immunotherapy of colonic carcinoma. PMID- 18387828 TI - Relationship between trabecular bone structure and articular cartilage morphology and relaxation times in early OA of the knee joint using parallel MRI at 3 T. AB - OBJECTIVE: To evaluate trabecular bone structure in relationship with cartilage parameters in distal femur and proximal tibia of the human knee at 3Tesla (3T) using high-resolution magnetic resonance imaging (MRI) with parallel imaging. METHOD: Sixteen healthy controls and 16 patients with mild osteoarthritis (OA) were studied using a 3T magnetic resonance (MR) scanner and an eight-channel phased-array knee coil. Axial 3D GeneRalized Autocalibrating Partially Parallel Acquisition (GRAPPA)-based phase cycled Fast Imaging Employing Steady State Acquisition (FIESTA-c) images were acquired in order to quantify the trabecular bone structure. For assessing cartilage morphology (thickness, volume), sagittal high-resolution 3D spoiled gradient echo (SPGR) images were acquired. In a subset of the subjects, sagittal images were acquired for measuring T1rho and T2 relaxation times, using 3D T1rho and T2 mapping techniques. RESULTS: Good measurement reproducibility was observed for bone parameters, the coefficients of variations (CVs) ranging from 1.8% for trabecular number (app. Tb.N) to 5.5% for trabecular separation (app. Tb.Sp). Significant differences between control and OA groups were found for bone volume fraction bone volume over total volume (app. BV/TV) and app. Tb.Sp in all compartments. Significantly increased values in T1rho and T2 were demonstrated in OA patients compared with controls at the femur, but not at the tibia. T1rho was negatively correlated with app. BV/TV, app. Tb.N and app. Tb.Sp both at the medial femoral condyle (MFC) and lateral tibia (LT), while T2 was only correlated at the LT. Also, medial tibia (MT) T1rho was negatively correlated with app. BV/TV (R(2)=-0.49, P<0.05) and app. Tb.N (R(2)=-0.42, P<0.05) from the opposite side of lateral femoral condyle (LFC). Significant correlations were found between trabecular bone parameters and cartilage thickness and normalized volume, mainly at LT, tibia (T) and femur (F). CONCLUSION: At this early stage of OA, an overall decrease in bone structure parameters and an increase in cartilage parameters (T1rho, T2) were noticed in patients. Trabecular bone structure correlated with articular cartilage parameters suggesting that loss of mineralized bone is associated with cartilage degeneration. PMID- 18387830 TI - Expression of glutamyl aminopeptidase by osteogenic induction in rat bone marrow stromal cells. AB - Glutamyl aminopeptidase (GluAP, EC 3.4.11.7, ENPEP) is a 130-kDa homodimeric zinc metallopeptidase which specifically cleaves the N-terminal glutamate or aspartate residue of peptidic substrates such as cholecystokinin-8 or angiotensin (Ang) II, in vitro. We used a DNA microarray hybridization (Genechip Rat Expression Array 230A, Affymetrix Inc., Santa Clara, CA, USA) to demonstrate that GluAP was upregulated in osteogenic induced rat bone marrow stromal cells (BMSCs). To compare the expression of GluAP in the osteogenic differentiation and non osteogenic differentiation of rat BMSCs in vitro, the cells were osteogenic induced in vitro. We also performed an MTT assay, alkaline phosphatase assay, alizarin red staining, and an immunohistochemical analysis to determine the osteogenic differentiation of BMSCs. The expression of GluAP was examined by real time polymerase chain reaction (PCR). The real-time PCR results showed that GluAP was upregulated in osteogenic differentiated BMSCs in vitro, suggesting that GluAP may be correlated with the osteogenic differentiation of BMSCs. PMID- 18387831 TI - Regulatory T cell-derived interleukin-10 limits inflammation at environmental interfaces. AB - The regulatory T (Treg) cells restrain immune responses through suppressor function elaboration that is dependent upon expression of the transcription factor Foxp3. Despite a critical role for Treg cells in maintaining lympho myeloid homeostasis, it remains unclear whether a single mechanism or multiple mechanisms of Treg cell-mediated suppression are operating in vivo and how redundant such mechanisms might be. Here we addressed these questions by examining the role of the immunomodulatory cytokine IL-10 in Treg cell-mediated suppression. Analyses of mice in which the Treg cell-specific ablation of a conditional IL-10 allele was induced by Cre recombinase knocked into the Foxp3 gene locus showed that although IL-10 production by Treg cells was not required for the control of systemic autoimmunity, it was essential for keeping immune responses in check at environmental interfaces such as the colon and lungs. Our study suggests that Treg cells utilize multiple means to limit immune responses. Furthermore, these mechanisms are likely to be nonredundant, in that a distinct suppressor mechanism most likely plays a prominent and identifiable role at a particular tissue and inflammatory setting. PMID- 18387832 TI - Constitutive crosspresentation of tissue antigens by dendritic cells controls CD8+ T cell tolerance in vivo. AB - Immature dendritic cells (DCs) sample tissue-specific antigens (TSAs) and process them for "crosspresentation" via major histocompatibility complex (MHC) class I and II molecules. Findings with adoptively transferred T cell receptor (TCR) transgenic CD8+ T cells in transgenic mice expressing model TSA indicate that this process contributes to tolerance induction of CD8+ T cells, a phenomenon termed "crosstolerance." However, up to now it has been unknown whether "crosstolerance" can also control autoimmune T cells specific for physiological nontransgenic TSA. Here, we showed that a DC-specific deficiency in uptake of apoptotic material inhibits crosspresentation in vivo. This defect allowed the accumulation of fully functional autoreactive CD8+ T cells that could be activated for autoimmune attack in peripheral lymphoid organs. Thus, our data demonstrate the importance of crosstolerance induction by DCs as a vital instrument for controlling self-reactive T cells from the peripheral repertoire and preventing autoimmune disease. PMID- 18387834 TI - Lack of the GTPase RHO-4 in Neurospora crassa causes a reduction in numbers and aberrant stabilization of microtubules at hyphal tips. AB - The multinucleate hyphae of the filamentous ascomycete fungus Neurospora crassa grow by polarized hyphal tip extension. Both the actin and microtubule cytoskeleton are required for maximum hyphal extension, in addition to other vital processes. Previously, we have shown that the monomeric GTPase encoded by the N. crassa rho-4 locus is required for actin ring formation during the process of septation; rho-4 mutants lack septa. However, other phenotypic aspects of the rho-4 mutant, such as slow growth and cytoplasmic bleeding, led us to examine the hypothesis that the microtubule (MT) cytoskeleton of the rho-4 mutant was affected in morphology and dynamics. Unlike a wild-type strain, the rho-4 mutant had few MTs and these few MTs originated from nuclear spindle pole bodies. rho-4 mutants and rho-4 strains containing a GTP-locked (activated) rho-4 allele showed a reduction in numbers of cytoplasmic MTs and microtubule stabilization at hyphal tips. Strains containing a GDP-biased (negative) allele of rho-4 showed normal numbers of MTs and minor effects on microtubule stabilization. An examination of nuclear dynamics revealed that rho-4 mutants have large, and often, stretched or broken nuclei. These observations indicate that RHO-4 plays important roles in regulating both the actin and MT cytoskeleton, which are essential for optimal hyphal tip growth and in nuclear distribution and morphology. PMID- 18387833 TI - The PI-3kinase pathway in hematopoietic stem cells and leukemia-initiating cells: a mechanistic difference between normal and cancer stem cells. AB - The identification of cancer stem cells in leukemia, breast, brain, colon, and other cancers suggests that many tumors are maintained by stem cells in much the same way as normal tissues are maintained. Because cancer stem cells share remarkable phenotypic and functional similarities with normal stem cells, it may be difficult to identify therapeutic approaches to kill cancer stem cells without killing the normal stem cells in the same tissue. Yet in certain tissues, like the hematopoietic system and gut epithelium, this will be critical as regenerative capacity in these tissues is acutely required for life. Components of the PI-3kinase pathway, including Akt, mTor and FoxO are critical regulators of both normal stem cell function and tumorigenesis. Intriguingly, inactivation of some pathway components, like Pten, has opposite effects on normal hematopoietic stem cells (HSCs) and leukemia-initiating cells. This raises the possibility that drugs targeting this pathway could be more effective at eliminating cancer stem cells while being less toxic against normal stem cells. PMID- 18387835 TI - Sam So Eum, a herb extract, as the remedy for allergen-induced asthma in mice. AB - We studied administering Sam So Eum (SSE) as a herbal medicine to treat asthma in mice and we discussed the mechanism of restoring the immuno-modulating cytokines such as IL-10 and IFN-gamma. The mice treated with SSE did not show any significant variation in their body weight and they looked very similar to the controlled ones. The SSE-treated mice showed reduced levels of airway responsiveness to methacholine, and these levels were initially elevated by the induction of asthma compared to the control group. The SSE elevated production level of IFN-gamma, which was down-regulated upon induction of asthma. This result implies that SSE can change the Th1/Th2 ratio through Th1-skewing reactions, and that SSE can decrease airway hyperresponsiveness by changing the Th1/Th2 ratio. The treatment with SSE also restored the IL-10 level to that of the naive condition. This means that SSE reduced the airway inflammation through this pathway. The ovalbumin (OVA)-specific antibody (total Ig) production in the serum was also decreased upon SSE treatment. Prednisolone (PD) was used as positive control. The effectiveness of SSE was almost the same as that of PD. These results suggest the possibility of using SSE for the treatment of patients with asthma, and its therapeutic efficacy involves restoring the IFN-gamma and IL 10 levels. PMID- 18387836 TI - Changes of gill and hemocyte-related bio-indicators during long term maintenance of the vent mussel Bathymodiolus azoricus held in aquaria at atmospheric pressure. AB - The deep-sea hydrothermal vent mussel Bathymodiolus azoricus has been the subject of several studies aimed at understanding the physiological adaptations that vent animals have developed in order to cope with the particular physical and chemical conditions of hydrothermal environments. In spite of reports describing successful procedures to maintain vent mussels under laboratory conditions at atmospheric pressure, few studies have described the mussel's physiological state after a long period in aquaria. In the present study, we investigate changes in mucocytes and hemocytes in B. azoricus over the course of several months after deep-sea retrieval. The visualization of granules of mucopolysaccharide or glycoprotein was made possible through their inherent auto-fluorescent property and the Alcian blue-Periodic Acid Schiff staining method. The density and distribution of droplets of mucus-like granules was observed at the ventral end of lamellae during acclimatization period. The mucus-like granules were greatly reduced after 3 months and nearly absent after 6 months of aquarium conditions. Additionally, we examined the depletion of endosymbiont bacteria from gill tissues, which typically occurs within a few weeks in sea water under laboratory conditions. The physiological state of B. azoricus after 6 months of acclimatization was also examined by means of phagocytosis assays using hemocytes. Hemocytes from mussels held in aquaria up to 6 months were still capable of phagocytosis but to a lesser extent when compared to the number of ingested yeast particles per phagocytic hemocytes from freshly collected vent mussels. We suggest that the changes in gill mucopolysaccharides and hemocyte glycoproteins, the endosymbiont abundance in gill tissues and phagocytosis are useful health criteria to assess long term maintenance of B. azoricus in aquaria. Furthermore, the laboratory set up to which vent mussels were acclimatized is an applicable system to study physiological reactions such as hemocyte immunocompetence even in the absence of the high hydrostatic pressure found at deep-sea vent sites. PMID- 18387837 TI - Isolation of lactoferrin from milk of different species: calorimetric and antimicrobial studies. AB - Lactoferrin (LF) is an iron-binding glycoprotein found in different biological fluids of mammals and in neutrophils. It has been proposed to be involved in many functions, including protection from pathogens. In this work, purification of lactoferrin using an ion-exchange chromatography (SP-Sepharose) was attempted for the milk of the following animals: sheep (Ovis aries), goat (Capra hircus), camel (Camelus bactrianus), alpaca (Lama pacos), elephant (Elephas maximus) and grey seal (Halichoerus grypus), as well as human (Homo sapiens). Lactoferrin was identified in all the milks apart from that from grey seal. The thermal stability of the purified lactoferrins, in their native and iron-saturated forms, was studied by differential scanning calorimetry (DSC). Maximum temperature, onset temperature and enthalpy change of denaturation were higher when lactoferrins were saturated with iron than in their native form, indicating an increase in the stability of the protein structure upon iron-binding. Human lactoferrin was found to be the most heat-resistant and the other lactoferrins presented different degrees of thermoresistance, that of elephant being the least resistant. The antimicrobial activity of the different isolated lactoferrins was investigated against Escherichia coli 0157:H7. The minimal inhibitory concentrations (MICs) were determined by measuring the absorbance at 620 nm. The minimum bactericidal concentrations (MBCs) were also measured and it was found that camel lactoferrin was the most active lactoferrin against E. coli 0157:H7, whereas alpaca and human lactoferrins were the least active. PMID- 18387838 TI - Single exon deletions in the PAH gene in Polish PKU-patients. PMID- 18387840 TI - Convenient ultrasound mediated synthesis of substituted pyrazolones under solvent free conditions. AB - Smooth condensation of hydrazine derivatives with various beta-keto esters was performed under solvent-free conditions by using ultrasound irradiation to facilitate the formation of pyrazolone derivatives in good to excellent amounts within very short time periods. PMID- 18387839 TI - Effects of an oral superoxide dismutase enzyme supplementation on indices of oxidative stress, proviral load, and CD4:CD8 ratios in asymptomatic FIV-infected cats. AB - This study was designed to test the effect of antioxidant supplementation on feline immunodeficiency virus (FIV)-infected felines. Six acutely FIV-infected cats (> or =16 weeks post-inoculation) were given a propriety oral superoxide dismutase (SOD) supplement (Oxstrin; Nutramax Laboratories) for 30 days. Following supplementation, the erythrocyte SOD enzyme concentration was significantly greater in the supplemented FIV-infected group than the uninfected control group or the unsupplemented FIV-infected group. The CD4+ to CD8+ ratio increased significantly (0.66-0.88) in the SOD supplemented FIV-infected cats but not in the unsupplemented FIV-infected cats. Proviral load and reduced glutathione (GSH) levels in leukocyte cell types did not change significantly following supplementation. Antioxidant supplementation resulted in an increase in SOD levels, confirming the oral bioavailability of the compound in FIV-infected cats. This result warrants further investigation with trials of antioxidant therapy in FIV-infected cats that are showing clinical manifestations of their disease, as well as in other feline patients where oxidative stress likely contributes to disease pathogenesis, such as diabetes mellitus and chronic renal failure. PMID- 18387841 TI - Bellini CM, et al. "Loss in mechanical contact of cementless acetabular prostheses due to postoperative weight bearing: a biomechanical model" [Med. Eng. Phys. 29(2) (2007) 175-181]. PMID- 18387842 TI - Greater motor improvement in right hemibody Parkinson's patients after dopaminergic medications. AB - Due to motor and neuropathological asymmetries, Parkinson's disease (PD) patients with right hemibody onset (RHO) of symptoms may experience greater benefit from dopaminergic enhancing medications, relative to patients with left hemibody onset (LHO). We investigated this possibility by measuring UPDRS scores of 20 PD patients with LHO and 13 patients with RHO, both prior and subsequent to taking dopaminergic enhancing medications. Following treatment, the motor score from the UPDRS improved for both groups of patients. However, PD patients with RHO experienced greater improvement in motor functioning following administration of dopaminergic enhancing medications than those with LHO. PMID- 18387843 TI - Lack of evidence for association of a parkin promoter polymorphism with early onset Parkinson's disease in a Chinese population. AB - Mutations in parkin are a common cause of early-onset autosomal recessive Parkinson's disease (PD). A single nucleotide polymorphism in the parkin promoter (rs9347683, -258T/G) has been reported to be associated with PD and shown to functionally affect gene transcription in luciferase reporter assays. In addition, homozygosity for the minor allele of rs9347683 may significantly reduce the age of onset of PD. We investigated the polymorphism in a cohort of cases with early-onset PD previously excluded for mutations in PD associated loci. We did not observe any differences in allele or genotype frequencies between the cases and the controls and there was no evidence for an effect on age of disease onset. Our results do not support a role for this variant in early-onset PD. PMID- 18387845 TI - Is MutaMouse insensitive to clastogens? AB - Several studies suggest that MutaMouse is insensitive to clastogens, including the accompanying paper by Mahabir et al., which describes a study with bleomycin, camptothecin, m-AMSA (4'-(9-acridinylamino)-methanesulfon-m-anisidide) and its ortho-analogue, o-AMSA (4'-(9-acridinylamino)-methanesulfon-o-anisidide). Only camptothecin was clastogenic in MutaMouse and none of these four compounds induced mutations at the lacZ locus. However, to improve exposure, dose range finding studies were performed in CD2F1 mice, the parental strain of MutaMouse. Male CD2F1 mice (n=3) were treated with bleomycin (25-100 mg/kg bw, p.o. and i.p.), camptothecin (1-10 mg/kg bw p.o.) and m-AMSA (10-50mg/kg bw p.o. and 1-5 mg/kg bw i.p.) for 5 days and blood was sampled on day 3 and/or day 6 for analysis by flow cytometry to determine % MN-RETs. Camptothecin (1 mg/kg bw, day 6) induced a 3.6-fold increase in % MN-RET (P<0.05) but was toxic at higher doses. All day-3 camptothecin samples were positive (P<0.05). Bleomycin was negative when administered p.o. but positive at all doses on both days when given i.p. (P<0.05) whereas m-AMSA was negative when given i.p. or orally. Based on these results, male MutaMouse mice (5 per group) were dosed daily with bleomycin (50 mg/kg bw) for 5 days or with camptothecin (5 mg/kg bw) for 2 days. Peripheral blood was sampled 24 h after the final dose in each group and tissues were sampled 37 days later. Both compounds induced significant increases in % MN-RET, but only bleomycin induced a significant increase in MF (6-fold in liver, 4.5 fold in kidney and 2-fold in lung) compared with the untreated control. These studies support the view that MutaMouse is insensitive to compounds where the genotoxic mechanism of action is predominantly clastogenesis, but demonstrates that the peripheral blood micronucleus test is a useful adjunct to the transgenic gene-mutation assay. PMID- 18387846 TI - Detecting genotoxic effects of potential clastogens: an in vivo study using the transgenic lacZ plasmid and the MutaMouse model. AB - In the present paper the capacity of the pUR288 plasmid mouse model and the MutaMouse model to detect the clastogens bleomycin, m-AMSA, o-AMSA and camptothecin, was investigated. Ethylnitrosourea (ENU) served as a positive control, methylcellulose as a negative control. Only bleomycin induced a slight but significant increase in lacZ mutant frequency (MF) in bone marrow of pUR288 plasmid mice. Exposure to the other compounds did not result in an increase in the MF in bone marrow and liver in both mouse models. For the MutaMouse this result was expected, for the plasmid mouse an increase in MF after clastogen exposure was expected. The positive control ENU induced statistically significant increases in MF compared with the negative control in both models and in both tissues analyzed. Hybridisation of DNA of mutant colonies derived from plasmid mice with labelled total mouse DNA (Hybridisation Assay) demonstrated an increase in the percentage of colonies hybridised with total mouse DNA as compared with the negative control, which suggests that there was indeed a biological response associated with treatment. The latter results indicate that the plasmid mouse assay may still be a promising model for the detection of clastogens. PMID- 18387847 TI - Two-dimensional near-infrared correlation spectroscopy study the methanol in acidic pH region. AB - The effect of pH on the structure of methanol was investigated by FT-NIR (Fourier transform near-infrared) spectroscopy and generalized two-dimensional (2D) correlation spectroscopy. pH perturbed 2D correlation spectra are calculated for the spectra in the 5,500-4,000, 7,500-5,500 cm(-1) regions at different pH values. We observed that the stretching of CH(3) was shifted because of the direct interaction of the CH(3) group of methanol with the OH group of water, the change of free OH is more sensitive to pH than the cyclic dimmer and CH.O. PMID- 18387844 TI - New insights into transcriptional regulation by H-NS. AB - H-NS, a nucleoid-associated DNA-binding protein of enteric bacteria, was discovered 35 years ago and subsequently found to exert widespread and highly pleiotropic effects on gene regulation. H-NS binds to high-affinity sites and spreads along adjacent AT-rich DNA to silence transcription. Preferential binding to sequences with higher AT-content than the resident genome allows H-NS to repress the expression of foreign DNA in a process known as 'xenogeneic silencing.' Counter-silencing by a variety of mechanisms facilitates the evolutionary acquisition of horizontally transferred genes and their integration into pre-existing regulatory networks. This review will highlight recent insights into the mechanism and biological importance of H-NS-DNA interactions. PMID- 18387848 TI - Factors involved in sleep satisfaction in the elderly. AB - BACKGROUND: In healthy aged subjects, subjective complaints of poor sleep are not as frequent as one would expect from the notable objective age-related sleep impairments. This discrepancy could depend on the sleep characteristics they require to feel satisfied about their sleep, which could be different from younger subjects. In order to verify this hypothesis, our study aims to identify changes in sleep satisfaction predictors as a function of age. METHODS: Sleep features, both quantitative (sleep latency, sleep duration, frequency and duration of night-time awakenings) and qualitative (calmness of sleep, ease of falling asleep, satisfaction with sleep, ease of awakening, freshness after awakening and sufficient sleep), as well as afternoon nap habits were investigated throughout a pre-arranged interview in healthy individuals: 117 elderly subjects (47 men, 70 women, 65-99 years) and 120 young adults (48 men, 72 women, 19-28 years). RESULTS: Quantitative sleep features worsen with age, whereas qualitative ones are equivalent or even improved in the aged group; only "calmness of sleep" decreases with age. Afternoon nap habits increase in elderly subjects, but they do not appear to be linked to sleep satisfaction. Predictors of sleep satisfaction differ between age groups: freshness after awakening but not frequency of night-time awakenings were relevant for elderly subjects, whereas the opposite was observed in young adults. CONCLUSIONS: These findings point out that sleep satisfaction is preserved in healthy elderly individuals despite the worsening of quantitative night-time sleep features, reflected by the increase of afternoon naps. This discrepancy could be explained by changes in sleep satisfaction determinants, towards which a spontaneous adaptive adjustment is likely to occur in aged subjects. In contrast with young adults, elderly subjects heed the freshness perceived after the awakening and pay little attention to frequent night-time awakenings. PMID- 18387851 TI - Towards gene therapy for Parkinson's disease. PMID- 18387850 TI - Safety and tolerability of intraputaminal delivery of CERE-120 (adeno-associated virus serotype 2-neurturin) to patients with idiopathic Parkinson's disease: an open-label, phase I trial. AB - BACKGROUND: There is an urgent need for therapies that slow or reverse the progression of Parkinson's disease (PD). Neurotrophic factors can improve the function of degenerating neurons and protect against further neurodegeneration, and gene transfer might be a means to deliver effectively these factors to the brain. The aim of this study was to assess the safety, tolerability, and potential efficacy of gene delivery of the neurotrophic factor neurturin. METHODS: In this phase I, open-label clinical trial, 12 patients aged 35-75 years with a diagnosis of PD for at least 5 years in accordance with the UK Brain Bank Criteria received bilateral, stereotactic, intraputaminal injections of adeno associated virus serotype 2-neurturin (CERE-120). The first six patients received doses of 1.3x10(11) vector genomes (vg)/patient, and the next six patients received 5.4x10(11) vg/patient. This trial is registered with ClinicalTrials.gov, number NCT00252850. FINDINGS: The procedure was well tolerated. Extensive safety monitoring in all patients revealed no clinically significant adverse events at 1 year. Several secondary measures of motor function showed improvement at 1 year; for example, a mean improvement in the off-medication motor subscore of the Unified Parkinson's Disease Rating Scale (UPDRS) of 14 points (SD 8; p=0.000121 [36% mean increase; p=0.000123]) and a mean increase of 2.3 h (2; 25% group mean increase; p=0.0250) in on time without troublesome dyskinesia were seen. Improvements in several secondary measures were not significant, including the timed walking test in the off condition (p=0.053), the Purdue pegboard test of hand dexterity (p=0.318), the reduction in off time (p=0.105), and the activities of daily living subscore (part II) of the UPDRS (p=0.080). (18)F-levodopa-uptake PET did not change after treatment with either dose of CERE-120. INTERPRETATION: The initial data support the safety, tolerability, and potential efficacy of CERE 120 as a possible treatment for PD; however, these results must be viewed as preliminary until data from blinded, controlled clinical trials are available. FUNDING: Ceregene; Michael J Fox Foundation for Parkinson's Research. PMID- 18387849 TI - Measuring malaria endemicity from intense to interrupted transmission. AB - The quantification of malaria transmission for the classification of malaria risk has long been a concern for epidemiologists. During the era of the Global Malaria Eradication Programme, measurements of malaria endemicity were institutionalised by their incorporation into rules outlining defined action points for malaria control programmes. We review the historical development of these indices and their contemporary relevance. This is at a time when many malaria-endemic countries are scaling-up their malaria control activities and reconsidering their prospects for elimination. These considerations are also important to an international community that has recently been challenged to revaluate the prospects for malaria eradication. PMID- 18387852 TI - Co-infusion of donor bone marrow with host mesenchymal stem cells treats GVHD and promotes vascularized skin allograft survival in rats. AB - We investigated the effect of autologous mesenchymal stem cells (MSC) on multiple unmodified donor bone marrow (BM) infusions and vascularized skin graft outcome. BM-derived rat MSC were examined for phenotype and function. MSC/MSC-conditioned medium suppressed IFN-gamma production by T cells and modified DC function. Infusions of MSC with one-time BM improved vascularized skin graft survival, while with one-two-times BM reversed graft versus host disease (GVHD). Mixed chimerism was enhanced in recipients given two-four-times BM with MSC infusions. Interestingly, four-times BM infusions with MSC delayed GVHD onset, reduced host tissue damage and enhanced vascularized skin allograft survival compared to four times BM alone. These data demonstrate that, the co-infusion of MSC with unmodified BM limit the toxicity of allogeneic BM transplantation, enhance mixed chimerism and improve vascularized skin graft survival. These findings provide insights for the development of autologous MSC-based BM transplantation and prevention of graft rejection or treatment of autoimmunity. PMID- 18387854 TI - Hypoalgesic effect of the transcutaneous electrical nerve stimulation following inguinal herniorrhaphy: a randomized, controlled trial. AB - We investigated the effect of transcutaneous electrical nerve stimulation (TENS) for inguinal herniorrhaphy postoperative pain control in a prospective, randomized, double-blinded, placebo-controlled study. Forty patients undergoing unilateral inguinal herniorrhaphy with an epidural anesthetic technique were randomly allocated to receive either active TENS or placebo TENS. Postoperative pain was evaluated using a standard 10-point numeric rating scale (NRS). Analgesic requirements were also recorded. TENS (100 Hz, strong but comfortable sensory intensity) was applied for 30 minutes through 4 electrodes placed around the incision twice, 2 and 4 hours after surgery. Pain was assessed before and after each application of TENS and 8 and 24 hours after surgery. In the group treated with active TENS, pain intensity was significantly lower 2 hours (P = .028), 4 hours (P = .022), 8 hours (P = .006), and 24 hours (P = .001) after the surgery when compared with the group that received placebo TENS. Active TENS also decreased analgesic requirements in the postoperative period when compared with placebo TENS (P = .001). TENS is thus beneficial for postoperative pain relief after inguinal herniorrhaphy; it has no observable side effects, and the pain reducing effect continued for at least 24 hours. Consequently, the routine use of TENS after inguinal herniorrhaphy is recommended. PERSPECTIVE: This study presents the hypoalgesic effect of high-frequency TENS for postoperative pain after inguinal herniorrhaphy. This may reinforce findings from basic science showing an opioid-like effect provided by TENS, given that high-frequency TENS has been shown to activate delta-opioid receptors. PMID- 18387853 TI - Affective analgesia following muscarinic activation of the ventral tegmental area in rats. AB - Cholinergic stimulation of dopamine neurons in the ventral tegmental area (VTA) underlies activation of the brain reward circuitry. Activation of this circuit is proposed to preferentially suppress the affective reaction to noxious stimulation. Vocalization afterdischarges (VADs) are a validated model of the affective response of rats to noxious tail shock. The antinociceptive action of the acetylcholine agonist carbachol microinjected into the VTA on VAD threshold was compared with its effect on the thresholds of other tail shock-elicited responses (VDS, vocalizations during shock; SMR, spinal motor reflexes). Whereas VADs are organized within the forebrain, VDSs and SMRs are organized at medullary and spinal levels of the neuraxis, respectively. Carbachol (1 microg, 2 microg, and 4 microg) injected into VTA produced dose-dependent increases in VAD and VDS thresholds, although increases in VAD threshold were significantly greater than increases in VDS threshold. Administration of carbachol into VTA failed to elevate SMR threshold. Elevations in vocalization thresholds produced by intra VTA carbachol were reversed in a dose-dependent manner by local administration of the muscarinic receptor antagonist atropine sulfate (30 microg and 60 microg). These results provide the first demonstration of the involvement of the VTA in muscarinic-induced suppression of pain affect. PERSPECTIVE: Cholinergic activation of the brain reward circuit produced a preferential suppression of rats' affective reaction to noxious stimulation. The neurobiology that relates reinforcement to suppression of pain affect may provide insights into new treatments for pain and its associated affective disorders. PMID- 18387856 TI - The analgesic effect of paeoniflorin on neonatal maternal separation-induced visceral hyperalgesia in rats. AB - Paeoniflorin (PF) is one of the principle active ingredients of the root of Paeonia lactiflora Pall (family Ranunculaceae), a Chinese herb traditionally used to relieve pain, especially visceral pain. The present study aimed to investigate both the effect of PF on neonatal maternal separation-induced visceral hyperalgesia in rats and the mechanism by which such effect is exerted. A dose dependent analgesic effect was produced by PF (45, 90, 180, and 360 mg/kg i.p.). Centrally administered PF (4.5 mg/kg i.c.v) also produced a significant analgesic effect. The analgesic effect of PF (45 mg/kg i.p.) was maximal at 30 minutes after administration. Furthermore, it was found that nor-binaltorphimine (nor BNI, 3 mg/kg i.p.), dl-alpha-methyltyrosine (alpha-AMPT, 250 mg/kg i.p.), and yohimbine (3 mg/kg i.p.) could block the analgesic effect of PF (45 mg/kg i.p.). Time course determination of PF in brain nuclei showed that the maximal concentration of PF was 30 minutes after intraperitoneal administration of PF (180 mg/kg) in cerebral nuclei, involving the amygdala, hypothalamus, thalamus, and cortex. These data indicate that PF has an analgesic effect on visceral pain in rats with neonatal maternal separation and that this effect may be mediated by kappa-opioid receptors and alpha(2)-adrenoceptors in the central nervous system. PERSPECTIVE: This study demonstrates that PF has an analgesic effect on pain in visceral hyperalgesic rats. These results suggest that PF might be potentially useful in clinical therapy for irritable bowel syndrome as a pharmacological agent in alleviating visceral pain. PMID- 18387855 TI - Intrathecal rosiglitazone acts at peroxisome proliferator-activated receptor gamma to rapidly inhibit neuropathic pain in rats. AB - In this report, we demonstrate the transcription, expression, and DNA-binding properties of the peroxisome proliferator-activated receptor (PPAR)-gamma subtype of the peroxisome proliferator-activated nuclear receptor family to the spinal cord with real-time PCR, Western blot, and electrophoretic mobility shift assay. To test the hypothesis that activation of spinal PPAR-gamma decreases nerve injury-induced allodynia, we intrathecally administered PPAR-gamma agonists and/or antagonists in rats after transection of the tibial and common peroneal branches of the sciatic nerve. Single injection of either a natural (15-deoxy prostaglandin J2, 15d-PGJ2) or synthetic (rosiglitazone) PPAR-gamma agonist dose dependently decreased mechanical and cold hypersensitivity. These effects were maximal at a dose of 100 microg and peaked at approximately 60 minutes after injection, a rapid time course suggestive of transcription-independent mechanisms of action. Concurrent administration of a PPAR-gamma antagonist (bisphenol A diglycidyl ether, BADGE) reversed the effects of 15d-PGJ2 and rosiglitazone, further indicating a receptor-mediated effect. In animals without nerve injury, rosiglitazone did not alter motor coordination, von Frey threshold, or withdrawal response to a cool stimulus. Intraperitoneal and intracerebroventricular administration of PPAR-gamma agonists (100 microg) did not decrease mechanical and cold hypersensitivity, arguing against effects subsequent to diffusion from the intrathecal space. We conclude that ligand-induced activation of spinal PPAR gamma rapidly reverses nerve injury-induced mechanical allodynia. New or currently available drugs targeted at spinal PPAR-gamma may yield important therapeutic effects for the management of neuropathic pain. PERSPECTIVE: PPAR gamma receptor agonists such as rosiglitazone and pioglitazone are approved as insulin sensitizers by the United States Food and Drug Administration. We demonstrate PPAR-gamma expression in the spinal cord and report that activation of these receptors inhibits allodynia. BBB-permeant PPAR-gamma agonists may yield important therapeutic effects for the management of neuropathic pain. PMID- 18387858 TI - Simultaneous determination of prednisolone, prednisone, cortisol, and cortisone in plasma by GC-MS: estimating unbound prednisolone concentration in patients with nephrotic syndrome during oral prednisolone therapy. AB - Individual variability of the pharmacokinetics of prednisolone based on the unbound concentration in plasma is of significant clinical consideration. The unbound concentrations of prednisolone were measured in 10 patients with nephrotic syndrome, two patients with systemic lupus erythematosus, and one patient with dermatomyositis by examining protein bindings of prednisolone on one or more occasions during prednisolone treatment. In this study, plasma concentrations of prednisolone, prednisone, cortisol, and cortisone were simultaneously analyzed by GC-MS by using stable isotope-labeled internal standards. Equilibrium dialysis was employed to accurately estimate the unbound fractions of prednisolone in plasma. The unbound fraction of prednisolone changed depending on plasma total prednisolone concentration and plasma albumin concentration. The unbound fraction of prednisolone (Y) is calculated: Y=( 0.0101x' + 0.0736) x + 10.23, where x' is the plasma albumin concentration and x is the total prednisolone concentration. The estimated concentrations of unbound prednisolone by using the above equation were in good agreement with the measured concentrations of unbound prednisolone. Since the protein binding of prednisolone did not change in the presence of prednisone (114.0 ng/ml), it appeared that prednisone produced from the therapeutic dose of prednisolone did not affect the unbound fraction of prednisolone. PMID- 18387859 TI - Eosinophilic ascites. PMID- 18387860 TI - In patients with ulcerative colitis, adsorptive depletion of granulocytes and monocytes impacts mucosal level of neutrophils and clinically is most effective in steroid naive patients. AB - BACKGROUND: The aetiology of ulcerative colitis is inadequately understood, and drug therapy has been empirical rather than based on sound understanding of disease aetiology. This has been a major factor for refractoriness and adverse drug effects as additional complications. However, ulcerative colitis by its very nature is exacerbated and perpetuated by inflammatory cytokines, which are released by peripheral granulocytes and monocytes as well. Additionally, active ulcerative colitis is often associated with elevated peripheral granulocytes and monocytes with activation behaviour and are found in vast numbers within the colonic mucosa. Hence, from the clinicopathologic viewpoint, granulocytes and monocytes are appropriate targets for therapy in ulcerative colitis. Based on this thinking, an Adacolumn has been developed for depleting excess granulocytes and monocytes by adsorption. METHODS: By colonoscopy, biopsy and histology, we investigated the impact of granulocyte and monocyte adsorption (GMA) on the mucosal level of granulocytes and monocytes in patients with active ulcerative colitis. Forty-five patients (26 steroid naive and 19 steroid-dependent), mean age 44.7 yr, were included. Twenty patients had total colitis and 25 had left sided colitis. Each patient was given up to 11 GMA sessions over 12 weeks. No patient received additional medications within 4 weeks (steroid) to 8 weeks (other immunosuppressants) prior to entry or during the GMA course. Colonoscopy together with biopsy was done at entry and within 2 weeks after the last GMA session. RESULTS: At entry, the mean clinical activity index was 12.6; range 10 16. A total of 400 colonic biopsies were examined, which revealed massive infiltration of the colonic mucosa by granulocytes, and GMA was associated with striking reduction of granulocytes in the mucosa. At week 12, 33 of 45 patients (73.3%, P<0.01) had achieved clinical remission (the mean clinical activity index stearate (740) > palmitate (590) > oleate (83) > linoleate (65). In general, cardioprotective FAs (e.g., oleate) increased expression of genes promoting FA oxidation to a greater extent than cardiotoxic FAs (e.g., palmitate), whereas the latter induced markers of endoplasmic reticulum and oxidative stress. Subsequent RT-PCR analysis revealed distinct time and concentration-dependent effects of these FA species, in a gene-specific manner. For example, stearate- and palmitate-mediated ucp3 induction tended to be transient (i.e., initial high induction, followed by subsequent repression), whereas oleate-mediated induction was sustained. These findings may provide insight into why diets high in unsaturated FAs (e.g., oleate) are cardioprotective, whereas diets rich in saturated FAs (e.g., palmitate) are not. PMID- 18387888 TI - NHS should bring in measures to reduce its carbon footprint, BMA says. PMID- 18387889 TI - HIV study yields unexpected preliminary efficacy results. PMID- 18387890 TI - FDA approves levoleucovorin. PMID- 18387893 TI - Government says patient safety improved 1% annually for 2000-05. PMID- 18387894 TI - Conversion to insulin devices in the inpatient setting. PMID- 18387895 TI - Caution in prescribing antidepressants for patients with cancer. PMID- 18387896 TI - Billing and reimbursement for intrathecal opiate therapy. PMID- 18387897 TI - Patient assessment tool for pharmacists. PMID- 18387898 TI - Current and future treatment modalities for Clostridium difficile-associated disease. AB - PURPOSE: Current and future treatment modalities for Clostridium difficile associated disease (CDAD) are reviewed. SUMMARY: C. difficile, an anaerobic, spore-forming, gram-positive [corrected] rod, is the enteric pathogen most frequently identified in patients with antibiotic-associated, nosocomially acquired diarrhea. Infection can lead to severe gastrointestinal illness which can develop into pseudomembranous colitis. Recent outbreaks in North America involved more virulent C. difficile strains, more severe infections, and more complicated treatment courses. Because of the potential for increased toxin associated damage due to increased toxin exposure time, CDAD treatment often involves cessation of the inciting antibiotic, C. difficile-targeted antibiotic therapy, electrolyte normalization, fluid replacement, and antimotility agent avoidance. First-line therapy for CDAD is treatment with the antibiotic metronidazole. Vancomycin is often used in more severe cases and for treatment resistant organisms. Treatment regimens may also include probiotics, bile-acid sequestrants, and, in limited cases, intravenous immunoglobulin (IVIG). Alternative treatments for refractory and persistent CDAD include intracolonic vancomycin, nitazoxanide, rifaximin, IVIG, and probiotics. Several target proteins have been proposed for C. difficile vaccine production, including the flagellar cap protein FliD, flagellin FLiC, a cell wall protein (Cwp) (comprising amino- and carboxyl-terminal domains), a protease Cwp 84, and toxins A and B. Rarely, pseudomembranous colitis, a severe complication of CDAD, must be treated through surgical intervention. CONCLUSION: CDAD is a major concern for health care systems and clinicians. New diagnostic tests with increased sensitivity for detecting CDAD with a short turnaround time are necessary for early treatment and prevention. Continued research for more effective treatments and vaccine development for CDAD is also needed. PMID- 18387899 TI - Prazosin for treatment of nightmares related to posttraumatic stress disorder. AB - PURPOSE: The efficacy of prazosin for the treatment of posttraumatic stress disorder (PTSD)-related nightmares is reviewed. SUMMARY: PTSD is an anxiety disorder that can occur after experiencing or witnessing a life-threatening event, such as military combat, natural disasters, terrorist attacks, serious accidents, or violent personal assaults. The event that induced PTSD is often relived through nightmares or flashbacks. Sleep disturbances affect approximately 70% of patients with PTSD. Several medications have been evaluated for reducing PTSD-related nightmares, with limited success. Prazosin is a centrally and peripherally acting alpha(1)-adrenergic antagonist whose mechanism of action, favorable adverse-effect profile, and low cost make it a promising agent for the treatment of PTSD. To date, two case reports, two chart reviews, three open-label trials, and two placebo-controlled trials have been published documenting the efficacy and safety of prazosin in the treatment of PTSD-related nightmares. Therapy with prazosin resulted in a reduction in nightmares in patients with both combat- and noncombat-related trauma. A therapeutic benefit occurred with prazosin dosages as low as 1 mg daily, and suppression of nightmare symptoms occurred within one week of prazosin initiation. The most frequently reported adverse event was orthostatic hypotension. The variability in the populations studied (e.g., combat, noncombat, recent traumatic experiences) leaves additional unanswered questions that must be addressed in large, randomized, controlled trials. CONCLUSION: Prazosin appears to be a promising and well-tolerated agent for the management of PTSD-related nightmares. Further well-designed trials are warranted to establish its place in the treatment of PTSD. PMID- 18387900 TI - Improving pain management for pediatric patients undergoing nonurgent painful procedures. AB - PURPOSE: The development of a topical analgesia protocol to improve pain management for pediatric patients undergoing nonurgent painful procedures is described. SUMMARY: Leadership from the departments of pediatrics, neonatology, obstetrics and gynecology, nursing, pharmacy, child life, and phlebotomy were chosen to develop and implement a new protocol for topical analgesia use for nonurgent painful procedures in pediatric patients. A review of the published literature on pain management in neonates, infants, children, and adolescents led to the replacement of lidocaine 2.5%-prilocaine 2.5% with liposomal lidocaine 4% topical cream on the formulary. In addition to topical analgesia, psychological and physical methods that enable children to cope successfully with anxiety provoking and painful experiences were included as part of the education portion of implementation. Child life staff educated other staff, patients, and their parents on pain management techniques, including deep breathing, imagery, and the use of distraction tools. The protocol was transcribed onto preprinted prescriber order forms, which were made available to all pediatric inpatient units, the pediatric emergency department, and the pediatric ambulatory care clinic. A separate form was developed for neonatology. Data from before and after protocol implementation were collected and assessed. Only pediatric patients admitted to inpatient units or seen in ambulatory care clinics were included in the evaluation. The percentage of patients undergoing nonurgent painful procedures treated with topical analgesia or dorsal penile block for circumcisions rose from 2% (preimplementation of protocol) to 92% (postimplementation) (p < 0.0001, chi square). CONCLUSION: A multidisciplinary approach to protocol development and implementation significantly increased compliance to a topical analgesia protocol for pediatric patients undergoing nonurgent painful procedures in a community medical center. PMID- 18387901 TI - Trial results: a fight for better patient care. PMID- 18387902 TI - NOTES: because we can or because we should? PMID- 18387903 TI - Takotsubo cardiomyopathy. PMID- 18387904 TI - Reversing motor vehicle injuries: a preventable tragedy. PMID- 18387905 TI - Non-human primate research: whither now? PMID- 18387906 TI - 'Acopia' and 'social admission' are not diagnoses: why older people deserve better. PMID- 18387907 TI - Bufale Spotting, part 9: assessing an economic evaluation. PMID- 18387908 TI - The conventional autopsy in modern medicine. AB - In many countries, including the UK, where relatives' consent is required, clinical autopsy rates (i.e. autopsies other than those required by law) have been declining since the 1950s. In the UK, even in teaching hospitals, the clinical autopsy rate has fallen to only 10% of deaths or less. At this rate of decline, clinical autopsies - and the pathologists who perform them - face extinction. The future practice of medicine will be blind to the many adverse consequences of clinical actions or omissions. The reasons for this decline are manifold and these have to be addressed if autopsy is to stand a chance of survival. The future of autopsy lies in promoting public support for autopsies, in some cases adapting the autopsy to address specific questions, thus making more effective use of information from autopsies. Only by ensuring that the next generation of doctors have experienced the powerful educational benefit of examining the body after death will the importance of autopsy to modern medicine be understood. PMID- 18387909 TI - Future career plans of a cohort of senior doctors working in the National Health Service. AB - OBJECTIVE: To report on the future career plans of senior doctors working in the NHS. DESIGN: Postal questionnaires. SETTING: All doctors who qualified in 1977 from all UK medical schools. MAIN OUTCOME MEASURES: Future plans and whether participants had any unmet needs for advice on how to put their future plans into effect. RESULTS: 25% definitely intended to continue with their current employment on the same basis until they retired; 75% hoped for change. A reduction in working hours was the most commonly desired change; but a substantial percentage also wanted changes in job content. 50% of respondents intended definitely (17%) or probably (33%) to work in the NHS to their normal retirement age; and 37% definitely (20%) or probably (17%) intended to retire early. 48% had made plans, in addition to the standard pension, to facilitate early retirement. The main factors given for considering early retirement were family reasons and wanting more time for leisure, a desire to maintain good health, excessive pressure of work, and disillusionment with NHS changes. A reduction in workload would be the greatest inducement to stay. 31% of respondents reported that they had unmet needs for advice about their future plans. Of these, about half were needs for advice about planning for retirement. CONCLUSIONS: Many senior NHS doctors would like to reduce their working hours. Less than a quarter definitely intend to work in the NHS to normal retirement age. Even for senior doctors, advice on career development is needed. PMID- 18387910 TI - Career destinations, job satisfaction and views of the UK medical qualifiers of 1977. AB - OBJECTIVE: To study the career destinations, job satisfaction and views of UK trained senior doctors. DESIGN: Postal questionnaire. SETTING: All doctors who qualified from all UK medical schools in 1977; and Department of Health employment data. MAIN OUTCOME MEASURES: Career destinations of medical qualifiers from 1977. RESULTS: 72% responded to the questionnaire. Using all available evidence, including that on non-responders, 76% of the cohort, comprising 77% of the men and 74% of the women, were working in the NHS 27 years after qualification. Approximately 18% were in medical jobs either overseas or outside the NHS. Of respondents in the NHS, 89% of men and 51% of women had full-time contracts. NHS doctors rated their job satisfaction highly, with a median score of 19.5 on a scale from 5 (very low satisfaction) to 25 (very high satisfaction). Satisfaction with time off for leisure was much lower, with a median score of 4.6 on a scale from 1 (low) to 10 (high). Of those in the NHS, 67% agreed that they worked longer hours than they thought they should; and 40% agreed that their working conditions were satisfactory. CONCLUSIONS: 27 years after qualification, the percentage of women who were working in the NHS was similar to that of men. Although these senior doctors had high levels of satisfaction with the content of their jobs, they were not so satisfied with their working hours and working conditions. Our results can be used as benchmarks, against which the career pathways and satisfaction levels of more recently qualified doctors can be compared. PMID- 18387911 TI - Splenic trauma during abdominal wall liposuction: a case report. AB - A 35-year-old woman collapsed 18 hours after undergoing abdominal wall liposuction. Abdominal CT scan revealed a punctured spleen. She underwent an emergency splenectomy and made an uneventful recovery. PMID- 18387912 TI - Jules Gavarret's Principes Generaux de Statistique Medicale. PMID- 18387913 TI - An academy of medical educators. PMID- 18387915 TI - Innate immunity to tropical theileriosis. AB - The intracellular protozoan parasite Theileria annulata causes a severe, and often fatal, disease of pure and cross-bred cattle in tropical and subtropical countries. The present review refers to the importance of innate immunity as far as it is known to date in this infectious disease. Specifically, macrophages and the mediators produced by these cells are outlined. In addition, the latest findings concerning cattle breed differences in susceptibility to T. annulata infection in relation to macrophage activation are discussed. PMID- 18387916 TI - Folimycin (concanamycin A) inhibits LPS-induced nitric oxide production and reduces surface localization of TLR4 in murine macrophages. AB - Lipopolysaccharide (LPS) is a major cell wall component of Gram-negative bacteria and signals through a receptor complex which consists of TLR4, MD-2 and CD14. LPS signaling in macrophages induces the production of many pro-inflammatory molecules, including nitric oxide (NO). In this study, we have shown that folimycin, a macrolide antibiotic and a specific inhibitor of vacuolar ATPase (V ATPase), inhibits LPS-induced NO production, but not TNFalpha production, in murine elicited peritoneal macrophages. However, folimycin did not affect interferon-gamma induced NO production. LPS-induced iNOS mRNA and protein expression and NF-kappaB activation were also inhibited by folimycin. Interestingly, folimycin-treated cells showed reduced surface expression of TLR4 molecules and dilated Golgi apparatus. These findings suggest that folimycin, by inhibiting V-ATPases, alters intra-Golgi pH, which in turn causes defective processing and reduced surface expression of TLR4 reducing the strength of LPS signaling in murine macrophages. PMID- 18387918 TI - Structural investigations into the interaction of hemoglobin and part structures with bacterial endotoxins. AB - An understanding of details of the interaction mechanisms of bacterial endotoxins (lipopolysaccharide, LPS) with the oxygen transport protein hemoglobin is still lacking, despite its high biological relevance. Here, a biophysical investigation into the endotoxin:hemoglobin interaction is presented which comprises the use of various rough mutant LPS as well as free lipid A; in addition to the complete hemoglobin molecule from fetal sheep extract, also the partial structure alpha chain and the heme-free sample are studied. The investigations comprise the determination of the gel-to-liquid crystalline phase behaviour of the acyl chains of LPS, the ultrastructure (type of aggregate structure and morphology) of the endotoxins, and the incorporation of the hemoglobins into artificial immune cell membranes and into LPS. Our data suggest a model for the interaction between Hb and LPS in which hemoglobins do not react strongly with the hydrophilic or with the hydrophobic moiety of LPS, but with the complete endotoxin aggregate. Hb is able to incorporate into LPS with the longitudinal direction parallel to the lipid A double-layer. Although this does not lead to a strong disturbance of the LPS acyl chain packing, the change of the curvature leads to a slightly conical molecular shape with a change of the three-dimensional arrangement from unilamellar into cubic LPS aggregates. Our previous results show that cubic LPS structures exhibit strong endotoxic activity. The property of Hb on the physical state of LPS described here may explain the observation of an increase in LPS mediating endotoxicity due to the action of Hb. PMID- 18387917 TI - Structure of a novel lipid A obtained from the lipopolysaccharide of Caulobacter crescentus. AB - Caulobacter crescentus CB15 is a dimorphic bacterium that is best known as a prokaryotic model for cell development. However, it is also being exploited in biotechnology, where the crystalline surface (S-layer) protein secretion system has been adapted for heterologous protein display or secretion. Because the S layer attaches to the cell surface via lipopolysaccharide (LPS) and since the LPS represents a potential endotoxin contaminant of recombinant proteins, the lipid A component was examined in detail. LPS was acid hydrolyzed to obtain crude lipid A, which was methylated and purified by HPLC. HPLC peak fractions were analyzed by mass spectrometry and nuclear magnetic resonance spectroscopy. The structure of the major lipid A of C. crescentus comprised the tetrasaccharide backbone alpha-D-GalpA-(1-->4)-beta-D-DAG-(1-->6)-alpha-D-DAG-(1-->1)-alpha-D-GalpA substituted with six fatty acids, and a molecular mass of 1875 (GalpA, galactopyranuronic acid; DAG, 2,3-diamino-2,3-dideoxyglucopyranose). No phosphate residues were detected. The major lipid A component had 12:0[3-O[Delta(5)-12:1(3 OH)]] and 12:0[3-O(Delta(5)-12:1)] fatty acyl chains at either the 3'- or the 2' positions of the distal subunit DAG B, and 12:0(3-OH) and 12:0[3,6-(OH)( 2)] fatty acyl chains at 3- and 2- positions of the reducing end subunit DAG A, respectively. In addition, several other variations in the structure were observed. The LPS was evaluated for TNF-alpha inducing activity and consistent with its unusual lipid A structure (relative to that of enteric bacteria), the activity was reduced by greater than 100-fold as compared to Escherichia coli ReLPS. This and other evidence suggests the potential application of this lipid A as a vaccine adjuvant or the suitability of Caulobacter displaying antigens for formulation of whole cell vaccines. PMID- 18387919 TI - Clearance of bacterial lipopolysaccharides and lipid A by the liver and the role of argininosuccinate synthase. AB - The liver is thought to be involved in the systemic clearance and detoxification of lipopolysaccharide (LPS). Argininosuccinate synthase (AS), a liver cytosolic urea cycle enzyme, has been found to bind to and inactivate LPS and lipid A. To elucidate the participation of AS in the clearance of LPS by liver and hepatocytes, we investigated the correlation between AS content and the removal of lipid A and LPS in vivo and in vitro, tracing levels of biological activity. A hepatotoxic model in which mice were injected with CCl(4) revealed a significant reduction in lipid A clearance along with liver failure on day 1; total body clearance was changed to 0.534 ml/min from 1.42 ml/min. AS content in liver concomitantly decreased to about half and AS leaked to blood at about 6 microg/ml. Total body clearance of i.v. injected AS was estimated at 0.083 ml/min, which predicted about 24-h leakage of AS after CCl(4) injection. The treatment also reduced the clearance of R-type LPSs to a lesser degree the larger its polysaccharide portion. S-type LPS, which has a large O-antigen polysaccharide, exhibited enhancement of clearance on CCl(4) treatment. When pretreated in vitro with AS and injected into normal mice, lipid A and R-type LPS showed a similar pattern of clearance of residual activities to the untreated forms, but S-type LPS exhibited enhancement of clearance. Comparison between different strains of mice revealed a correlation of AS content in liver and lipid A clearance, where the higher AS strain C3H/He mice showed a more rapid clearance than the lower AS strains C57BL/6 and BALB/c. Primary spheroid cultures of hepatocytes treated with 0.1 microM dexamethasone and 1 microM glucagon showed about a 2-fold increase in AS amount and a more rapid clearance of LPS from culture medium than untreated cells. These results suggest that AS in hepatocytes may be involved in the process of lipid A and LPS clearance and the extracellular leakage of AS may also participate in the systemic detoxification. PMID- 18387920 TI - The cover. Flora. PMID- 18387921 TI - A piece of my mind. Someone else's sister. PMID- 18387922 TI - HIV prevention studies yield mixed results. PMID- 18387923 TI - Global report highlights tobacco use, offers countermeasures for nations. PMID- 18387924 TI - Grapefruit compound battles hepatitis C. PMID- 18387925 TI - Effect of aspirin use on thiazolidinediones and cardiovascular events. PMID- 18387926 TI - Sedation with dexmedetomidine vs lorazepam in mechanically ventilated patients. PMID- 18387927 TI - Sedation with dexmedetomidine vs lorazepam in mechanically ventilated patients. PMID- 18387928 TI - Sedation with dexmedetomidine vs lorazepam in mechanically ventilated patients. PMID- 18387929 TI - Bone mineral density and high-trauma fractures. PMID- 18387930 TI - Genetically guided choices for chemotherapy. PMID- 18387932 TI - Gene expression signatures, clinicopathological features, and individualized therapy in breast cancer. AB - CONTEXT: Gene expression profiling may be useful for prognostic and therapeutic strategies in breast carcinoma. OBJECTIVES: To demonstrate the value in integrating genomic information with clinical and pathological risk factors, to refine prognosis, and to improve therapeutic strategies for early stage breast cancer. DESIGN, SETTING, AND PATIENTS: Retrospective study of patients with early stage breast carcinoma who were candidates for adjuvant chemotherapy; 964 clinically annotated breast tumor samples (573 in the initial discovery set and 391 in the validation cohort) with corresponding microarray data were used. All patients were assigned relapse risk scores based on their respective clinicopathological features. Signatures representing oncogenic pathway activation and tumor biology/microenvironment status were applied to these samples to obtain patterns of deregulation that correspond with relapse risk scores to refine prognosis with the clinicopathological prognostic model alone. Predictors of chemotherapeutic response were also applied to further characterize clinically relevant heterogeneity in early stage breast cancer. MAIN OUTCOME MEASURES: Gene expression signatures and clinicopathological variables in early stage breast cancer to determine a refined estimation of relapse-free survival and sensitivity to chemotherapy. RESULTS: In the initial data set of 573 patients, prognostically significant clusters representing patterns of oncogenic pathway activation and tumor biology/microenvironment states were identified within the low-risk (log-rank P = .004), intermediate-risk (log-rank P = .01), and high-risk (log-rank P = .003) model cohorts, representing clinically important genomic subphenotypes of breast cancer. As an example, in the low-risk cohort, of 6 prognostically significant clusters, patients in cluster 4 had an inferior relapse-free survival vs patients in cluster 1 (log-rank P = .004) and cluster 5 (log-rank P = .03). Median relapse-free survival for patients in cluster 4 was 16 months less than for patients in cluster 1 (95% CI, 7.5-24.5 months) and 19 months less than for patients in cluster 5 (95% CI, 10.5-27.5 months). Multivariate analyses confirmed the independent prognostic value of the genomic clusters (low risk, P = .05; high risk, P = .02). The reproducibility and validity of these patterns of pathway deregulation in predicting relapse risk was established using related but not identical clusters in the independent validation cohort. The prognostic clinicogenomic clusters also have unique sensitivity patterns to commonly used cytotoxic therapies. CONCLUSIONS: These results provide preliminary evidence that incorporation of gene expression signatures into clinical risk stratification can refine prognosis. Prospective studies are needed to determine the value of this approach for individualizing therapeutic strategies. PMID- 18387933 TI - 21st-century primary care: new physician roles need new payment models. PMID- 18387934 TI - Newly approved does not always mean new and improved. PMID- 18387935 TI - The hope and fear of rimonabant. PMID- 18387936 TI - Use of gene signatures to improve risk estimation in cancer. PMID- 18387931 TI - Effect of rimonabant on progression of atherosclerosis in patients with abdominal obesity and coronary artery disease: the STRADIVARIUS randomized controlled trial. AB - CONTEXT: Abdominal obesity is associated with metabolic abnormalities and increased risk of atherosclerotic cardiovascular disease. However, no obesity management strategy has demonstrated the ability to slow progression of coronary disease. OBJECTIVE: To determine whether weight loss and metabolic effects of the selective cannabinoid type 1 receptor antagonist rimonabant reduces progression of coronary disease in patients with abdominal obesity and the metabolic syndrome. DESIGN, SETTING, AND PATIENTS: Randomized, double-blinded, placebo controlled, 2-group, parallel-group trial (enrollment December 2004-December 2005) comparing rimonabant with placebo in 839 patients at 112 centers in North America, Europe, and Australia. INTERVENTIONS: Patients received dietary counseling, were randomized to receive rimonabant (20 mg daily) or matching placebo, and underwent coronary intravascular ultrasonography at baseline (n = 839) and study completion (n = 676). MAIN OUTCOME MEASURES: The primary efficacy parameter was change in percent atheroma volume (PAV); the secondary efficacy parameter was change in normalized total atheroma volume (TAV). RESULTS: In the rimonabant vs placebo groups, PAV (95% confidence interval [CI]) increased 0.25% (-0.04% to 0.54%) vs 0.51% (0.22% to 0.80%) (P = .22), respectively, and TAV decreased 2.2 mm3 (-4.09 to -0.24) vs an increase of 0.88 mm3 (-1.03 to 2.79) (P = .03). In the rimonabant vs placebo groups, imputing results based on baseline characteristics for patients not completing the trial, PAV increased 0.25% ( 0.04% to 0.55%) vs 0.57% (0.29% to 0.84%) (P = .13), and TAV decreased 1.95 mm3 ( 3.8 to -0.10) vs an increase of 1.19 mm3 (-0.73 to 3.12) (P = .02). Rimonabant treated patients had a larger reduction in body weight (4.3 kg [-5.1 to -3.5] vs 0.5 kg [-1.3 to 0.3]) and greater decrease in waist circumference (4.5 cm [-5.4 to -3.7] vs 1.0 cm [-1.9 to -0.2]) (P < .001 for both comparisons). In the rimonabant vs placebo groups, high-density lipoprotein cholesterol levels increased 5.8 mg/dL (4.9 to 6.8) (22.4%) vs 1.8 mg/dL (0.9 to 2.7) (6.9%) (P < .001), and median triglyceride levels decreased 24.8 mg/dL (-35.4 to -17.3) (20.5%) vs 8.9 mg/dL (-14.2 to -1.8) (6.2%) (P < .001). Rimonabant-treated patients had greater decreases in high-sensitivity C-reactive protein (1.3 mg/dL [-1.7 to -1.2] [50.3%] vs 0.9 mg/dL [-1.4 to -0.5] [30.9%]) and less increase in glycated hemoglobin levels (0.11% [0.02% to 0.20%] vs 0.40% [0.31% to 0.49%]) (P < .001 for both comparisons). Psychiatric adverse effects were more common in the rimonabant group (43.4% vs 28.4%, P < .001). CONCLUSIONS: After 18 months of treatment, the study failed to show an effect for rimonabant on disease progression for the primary end point (PAV) but showed a favorable effect on the secondary end point (TAV). Determining whether rimonabant is useful in management of coronary disease will require additional imaging and outcomes trials, which are currently under way. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00124332. PMID- 18387937 TI - JAMA patient page. Pancreatitis. PMID- 18387938 TI - Anti-serum albumin domain antibodies for extending the half-lives of short lived drugs. AB - We have used phage display to isolate a range of human domain antibodies (dAbs) that bind to mouse, rat and/or human serum albumin (SA) and can be expressed at very high levels in bacterial, yeast or mammalian cell culture. In contrast to non-SA-binding dAbs, which have terminal half-lives of less than 45 min, the half lives of these 12 kDa 'AlbudAbs' can match the half-life of SA itself. To demonstrate the use of AlbudAbs for extending the half-lives of therapeutic drugs, we created a fusion of the interleukin-1 receptor antagonist (IL-1ra) with an AlbudAb. Soluble IL-1ra is potent inhibitor of IL-1 signalling that is approved for the treatment of rheumatoid arthritis but has a relatively short in vivo half-life. Here we show that although the AlbudAb/IL-1ra fusion has a similar in vitro potency, its in vivo efficacy can be dramatically improved due to its extended serum half-life. AlbudAbs could potentially be used to generate a range of long half-life versions of many different drugs in order to improve their dosing regimen and/or clinical effect. PMID- 18387939 TI - Transcatheter closure of a large abdominal aortic pseudoaneurysm with a septal occluder device. PMID- 18387940 TI - Management and 6-month outcomes in elderly and very elderly patients with high risk non-ST-elevation acute coronary syndromes: The Global Registry of Acute Coronary Events. AB - AIMS: To test the hypothesis that increasing age in patients presenting with high risk non-ST-segment elevation acute coronary syndromes (NSTE-ACS) does not adversely influence the benefit of an early invasive strategy on major adverse events at 6 months. METHODS AND RESULTS: We report clinical outcomes in young (<70), elderly (70-80), and very elderly (>80 years) patients with high-risk NSTE ACS enrolled in GRACE between 1999 and 2006. Six month data were available in 18 466 patients (27% elderly, 16% very elderly). Elderly and very elderly patients were less likely to receive evidence-based treatments at discharge and had a longer hospital stay (6 vs. 5 days). Angiography was performed more frequently in younger patients (67 vs. 33% in very elderly, 55% in elderly; P < 0.0001). Multiple logistic regression analysis confirmed the benefit of revascularization on the primary study endpoint (6-month stroke, death, myocardial infarction) in young [odds ratio (OR) 0.69, 95% confidence interval (CI) 0.56-0.86], elderly (0.60, 0.47-0.76), and very elderly (0.72, 0.54-0.95) patients. Revascularization was associated with reductions in 6-month mortality (OR 0.52, 95% CI 0.37-0.72 in young; 0.38, 0.26-0.54 in elderly; 0.68, 0.49-0.95 in very elderly). Stroke risk in hospital or at 6 months was not increased by revascularization. CONCLUSION: Following presentation with high-risk NSTE-ACS, an evidence-based approach to management was noted less frequently with advancing patient age. Angiography, in particular, was less likely to be undertaken. Revascularization, however, when performed, was associated with significant benefits at 6 months, independent of age, and did not increase risk of stroke. PMID- 18387941 TI - Assembly of heterohexameric trypanosome hexokinases reveals that hexokinase 2 is a regulable enzyme. AB - Glycolysis is essential to Trypanosoma brucei, the protozoan parasite that causes African sleeping sickness in humans and nagana in cattle. Hexokinase (HK), the first enzyme in glycolysis, catalyzes the phosphorylation of glucose to form glucose 6-phosphate. T. brucei harbors two HKs that are 98% identical at the amino acid level, T. brucei hexokinase 1 (TbHK1) and TbHK2. Recombinant TbHK1 (rTbHK1) has HK activity, whereas rTbHK2 does not. Unlike other eukaryotic HKs, TbHK1 is not subject to inhibition by ADP and glucose 6-phosphate. However, TbHK1 is inhibited by myristate, a critical fatty acid in T. brucei biology. We report here that rTbHKs, similar to authentic TbHK, form oligomers. Myristate dissociated these assemblies when incubated with either ATP or glucose. Furthermore, oligomer disruption was reversible by removal of myristate. Mixing of rTbHK1 and rTbHK2 monomers followed by reassembly yielded enzyme with an approximately 3-fold increase in specific activity compared with similarly treated rTbHK1 alone. Surprisingly, reassembly of rTbHK2 with an inactive rTbHK1 variant yielded an active HK, revealing for the first time that rTbHK2 is competent for HK activity. Finally, pyrophosphate inhibits active reassembled rTbHK2 oligomers but not oligomeric rTbHK1, suggesting that the two enzymes have distinct regulatory mechanisms. PMID- 18387942 TI - Disrupted RabGAP function of the p85 subunit of phosphatidylinositol 3-kinase results in cell transformation. AB - Rab proteins regulate vesicle fusion events during the endocytosis, recycling, and degradation of activated receptor tyrosine kinases. The p85alpha subunit of phosphatidylinositol 3-kinase has GTPase-activating protein activity toward Rab5 and Rab4, an activity severely reduced by a single point mutation (p85-R274A). Expression of p85-R274A resulted in increased platelet-derived growth factor receptor (PDGFR) activation and downstream signaling (Akt and MAPK) and in decreased PDGFR degradation. We now report that the biological consequences of p85-R274A expression cause cellular transformation as determined by the following: aberrant morphological phenotype, loss of contact inhibition, growth in soft agar, and tumor formation in nude mice. Immunohistochemistry shows that the tumors contain activated PDGFR and high levels of activated Akt. Coexpression of a dominant negative Rab5-S34N mutant attenuated these transformed properties. Our results demonstrate that disruption of the RabGAP function of p85alpha due to a single point mutation (R274A) is sufficient to cause cellular transformation via a phosphatidylinositol 3-kinase-independent mechanism partially reversed by Rab5-S34N expression. This critical new role for p85 in the regulation of Rab function suggests a novel role for p85 in controlling receptor signaling and trafficking through its effects on Rab GTPases. PMID- 18387944 TI - Structural and functional insights into the molecular mechanisms responsible for the regulation of pyruvate dehydrogenase kinase 2. AB - PDHK2 is a mitochondrial protein kinase that phosphorylates pyruvate dehydrogenase complex, thereby down-regulating the oxidation of pyruvate. Here, we present the crystal structure of PDHK2 bound to the inner lipoyl-bearing domain of dihydrolipoamide transacetylase (L2) determined with or without bound adenylyl imidodiphosphate. Both structures reveal a PDHK2 dimer complexed with two L2 domains. Comparison with apo-PDHK2 shows that L2 binding causes rearrangements in PDHK2 structure that affect the L2- and E1-binding sites. Significant differences are found between PDHK2 and PDHK3 with respect to the structure of their lipoyllysine-binding cavities, providing the first structural support to a number of studies showing that these isozymes are markedly different with respect to their affinity for the L2 domain. Both structures display a novel type II potassium-binding site located on the PDHK2 interface with the L2 domain. Binding of potassium ion at this site rigidifies the interface and appears to be critical in determining the strength of L2 binding. Evidence is also presented that potassium ions are indispensable for the cross-talk between the nucleotide- and L2-binding sites of PDHK2. The latter is believed to be essential for the movement of PDHK2 along the surface of the transacetylase scaffold. PMID- 18387943 TI - Phorbol 12-myristate 13-acetate-dependent protein kinase C delta-Tyr311 phosphorylation in cardiomyocyte caveolae. AB - Protein kinase Cdelta (PKCdelta) activation is generally attributed to lipid cofactor-dependent allosteric activation mechanisms at membranes. However, recent studies indicate that PKCdelta also is dynamically regulated through tyrosine phosphorylation in H(2)O(2)- and phorbol 12-myristate 13-acetate (PMA)-treated cardiomyocytes. H(2)O(2) activates Src and related Src-family kinases (SFKs), which function as dual PKCdelta-Tyr(311) and -Tyr(332) kinases in vitro and contribute to H(2)O(2)-dependent PKCdelta-Tyr(311)/Tyr(332) phosphorylation in cardiomyocytes and in mouse embryo fibroblasts. H(2)O(2)-dependent PKCdelta Tyr(311)/Tyr(332) phosphorylation is defective in SYF cells (deficient in SFKs) and restored by Src re-expression. PMA also promotes PKCdelta-Tyr(311) phosphorylation, but this is not associated with SFK activation or PKCdelta Tyr(332) phosphorylation. Rather, PMA increases PKCdelta-Tyr(311) phosphorylation by delivering PKCdelta to SFK-enriched caveolae. Cyclodextrin treatment disrupts caveolae and blocks PMA-dependent PKCdelta-Tyr(311) phosphorylation, without blocking H(2)O(2)-dependent PKCdelta-Tyr(311) phosphorylation. The enzyme that acts as a PKCdelta-Tyr(311) kinase without increasing PKCdelta phosphorylation at Tyr(332) in PMA-treated cardiomyocytes is uncertain. Although in vitro kinase assays implicate c-Abl as a selective PKCdelta-Tyr(311) kinase, PMA-dependent PKCdelta-Tyr(311) phosphorylation persists in cardiomyocytes treated with the c Abl inhibitor ST1571 and c-Abl is not detected in caveolae; these results effectively exclude a c-Abl-dependent process. Finally, we show that 1,2-dioleoyl sn-glycerol mimics the effect of PMA to drive PKCdelta to caveolae and increase PKCdelta-Tyr(311) phosphorylation, whereas G protein-coupled receptor agonists such as norepinephrine and endothelin-1 do not. These results suggest that norepinephrine and endothelin-1 increase 1,2-dioleoyl-sn-glycerol accumulation and activate PKCdelta exclusively in non-caveolae membranes. Collectively, these results identify stimulus-specific PKCdelta localization and tyrosine phosphorylation mechanisms that could be targeted for therapeutic advantage. PMID- 18387945 TI - Identification and functional analysis of phosphorylated tyrosine residues within EphA2 receptor tyrosine kinase. AB - EphA2 is a member of the Eph family of receptor tyrosine kinases. EphA2 mediates cell-cell communication and plays critical roles in a number of physiological and pathologic responses. We have previously shown that EphA2 is a key regulator of tumor angiogenesis and that tyrosine phosphorylation regulates EphA2 signaling. To understand the role of EphA2 phosphorylation, we have mapped phosphorylated tyrosines within the intracellular region of EphA2 by a combination of mass spectrometry analysis and phosphopeptide mapping using two-dimensional chromatography in conjunction with site-directed mutagenesis. The function of these phosphorylated tyrosine residues was assessed by mutational analysis using EphA2-null endothelial cells reconstituted with EphA2 tyrosine-to-phenylalanine or tyrosine-to-glutamic acid substitution mutants. Phosphorylated Tyr(587) and Tyr(593) bind to Vav2 and Vav3 guanine nucleotide exchange factors, whereas Tyr(P)(734) binds to the p85 regulatory subunit of phosphatidylinositol 3-kinase. Mutations that uncouple EphA2 with Vav guanine nucleotide exchange factors or p85 are defective in Rac1 activation and cell migration. Finally, EphA2 mutations in the juxtamembrane region (Y587F, Y593F, Y587E/Y593E), kinase domain (Y734F), or SAM domain (Y929F) inhibited ephrin-A1-induced vascular assembly. In addition, EphA2-null endothelial cells reconstituted with these mutants were unable to incorporate into tumor vasculature, suggesting a critical role of these phosphorylation tyrosine residues in transducing EphA2 signaling in vascular endothelial cells during tumor angiogenesis. PMID- 18387946 TI - Regiospecific O-methylation of naphthoic acids catalyzed by NcsB1, an O methyltransferase involved in the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin. AB - Neocarzinostatin, a clinical anticancer drug, is the archetypal member of the chromoprotein family of enediyne antitumor antibiotics that are composed of a nonprotein chromophore and an apoprotein. The neocarzinostatin chromophore consists of a nine-membered enediyne core, a deoxyaminosugar, and a naphthoic acid moiety. We have previously cloned and sequenced the neocarzinostatin biosynthetic gene cluster and proposed that the biosynthesis of the naphthoic acid moiety and its incorporation into the neocarzinostatin chromophore are catalyzed by five enzymes NcsB, NcsB1, NcsB2, NcsB3, and NcsB4. Here we report the biochemical characterization of NcsB1, unveiling that: (i) NcsB1 is an S adenosyl-L-methionine-dependent O-methyltransferase; (ii) NcsB1 catalyzes regiospecific methylation at the 7-hydroxy group of its native substrate, 2,7 dihydroxy-5-methyl-1-naphthoic acid; (iii) NcsB1 also recognizes other dihydroxynaphthoic acids as substrates and catalyzes regiospecific O-methylation; and (iv) the carboxylate and its ortho-hydroxy groups of the substrate appear to be crucial for NcsB1 substrate recognition and binding, and O-methylation takes place only at the free hydroxy group of these dihydroxynaphthoic acids. These findings establish that NcsB1 catalyzes the third step in the biosynthesis of the naphthoic acid moiety of the neocarzinostatin chromophore and further support the early proposal for the biosynthesis of the naphthoic acid and its incorporation into the neocarzinostatin chromophore with free naphthoic acids serving as intermediates. NcsB1 represents another opportunity that can now be exploited to produce novel neocarzinostatin analogs by engineering neocarzinostatin biosynthesis or applying directed biosynthesis strategies. PMID- 18387947 TI - Transforming growth factor beta up-regulates cysteine-rich protein 2 in vascular smooth muscle cells via activating transcription factor 2. AB - CRP2 (cysteine-rich protein) is a vascular smooth muscle cell (VSMC)-expressed LIM-only protein. CRP2 associates with the actin cytoskeleton and interacts with transcription factors in the nucleus to mediate smooth muscle cell gene expression. Using Csrp2 (gene symbol of the mouse CRP2 gene)-deficient mice, we previously demonstrated that an absence of CRP2 enhances VSMC migration and increases neointima formation following arterial injury. Despite its importance in vascular injury, the molecular mechanisms controlling CRP2 expression in VSMC are largely unknown. Transforming growth factor beta (TGFbeta), a key factor present in the vessel wall in the early phases of arterial response to injury, plays an important role in modulating lesion formation. Because both CRP2 and TGFbeta are mediators of VSMC responses, we examined the possibility that TGFbeta might regulate CRP2 expression. TGFbeta significantly induced CRP2 mRNA and protein expression in VSMCs. Promoter analysis identified a conserved cAMP responsive element (CRE)-like site (TAACGTCA) in the Csrp2 promoter that was critical for basal promoter activity and response to TGFbeta. Gel mobility shift assays revealed that mainly ATF2 bound to this CRE-like element, and mutation of the CRE sequences abolished binding. TGFbeta enhanced the activation of ATF2, leading to increased phospho-ATF2 levels within the DNA-protein complexes. Furthermore, ATF2-transactivated Csrp2 promoter activity and TGFbeta enhanced this activation. In addition, a phosphorylation-negative ATF2 mutant construct decreased basal and TGFbeta-mediated Csrp2 promoter activity. Our results show for the first time in VSMC that TGFbeta activates ATF2 phosphorylation and Csrp2 gene expression via a CRE promoter element. PMID- 18387948 TI - PATE gene clusters code for multiple, secreted TFP/Ly-6/uPAR proteins that are expressed in reproductive and neuron-rich tissues and possess neuromodulatory activity. AB - We report here syntenic loci in humans and mice incorporating gene clusters coding for secreted proteins each comprising 10 cysteine residues. These conform to three-fingered protein/Ly-6/urokinase-type plasminogen activator receptor (uPAR) domains that shape three-fingered proteins (TFPs). The founding gene is PATE, expressed primarily in prostate and less in testis. We have identified additional human PATE-like genes (PATE-M, PATE-DJ, and PATE-B) that co-localize with the PATE locus, code for novel secreted PATE-like proteins, and show selective expression in prostate and/or testis. Anti-PATE-B-specific antibodies demonstrated the presence of PATE-B in the region of the sperm acrosome and at high levels on malignant prostatic epithelial cells. The syntenic mouse Pate-like locus encompasses 14 active genes coding for secreted proteins, which are all, except for Pate-P and Pate-Q, expressed primarily in prostate and/or testis. Pate P and Pate-Q are expressed solely in placental tissue. Castration up-regulates prostate expression of mouse Pate-B and Pate-E, whereas testosterone ablates this induced expression. The sequence similarity between TFP/Ly-6/uPAR proteins that modulate activity of nicotinic acetylcholine receptors and the PATE (Pate)-like proteins stimulated us to see whether these proteins possess analogous activity. Pharmacological studies showed significant modulation of the nicotinic acetylcholines by the PATE-B, Pate-C, and Pate-P proteins. In concert with these findings, certain PATE (Pate)-like genes were extensively expressed in neuron rich tissues. Taken together, our findings indicate that in addition to participation of the PATE (Pate)-like genes in functions related to fertility and reproduction, some of them likely act as important modulators of neural transmission. PMID- 18387949 TI - Insulin effects on cardiac Na+/Ca2+ exchanger activity: role of the cytoplasmic regulatory loop. AB - Insulin can alter myocardial contractility, in part through an effect on the cardiac sarcolemmal Na(+)/Ca(2+) exchanger (NCX), but little is known about its mechanism of action. The large cytoplasmic domain (f-loop) of NCX is required for regulation by various intracellular factors, and we have shown previously that residues 562-679 are determinants of NCX inhibition by exchanger inhibitory peptide (XIP). Here we show that the same f-loop deletion eliminates the enhancement of NCX current by insulin, and we examine the signal pathways involved in the insulin response. NCX current (I(NCX)) was measured in freshly isolated or cultured (up to 48 h) adult guinea pig myocytes and in myocytes expressing canine NCX1.1 with the 562-679 f-loop deletion (NCX-(Delta562-679)) via adenoviral gene transfer. I(NCX) was recorded by whole-cell patch clamp as the Ni(2+)-sensitive current at 37 degrees C with intracellular Ca(2+) buffered. Insulin (1 microm) increased I(NCX) (at +80 mV) by 110 and 83% in fresh and cultured myocytes, respectively, whereas in myocytes expressing NCX-(Delta562 679) the response was eliminated (with 100 microm XIP included to suppress any native guinea pig I(NCX)). The insulin effect on I(NCX) was not inhibited by wortmannin, a nitric-oxide synthase inhibitor, or disruption of caveolae but was blocked by chelerythrine, implicating protein kinase C, but not phosphatidylinositol-3-kinase, in the mechanism. The insulin effect was also not additive with phosphatidylinositol-4,5-bisphosphate-induced activation of I(NCX). The finding that the 562-670 f-loop domain is implicated in both XIP and receptor mediated modulation of NCX highlights its important role in acute physiological or pathophysiological regulation of Ca(2+) balance in the heart. PMID- 18387950 TI - Structural signatures and membrane helix 4 in GLUT1: inferences from human blood brain glucose transport mutants. AB - Exon IV of SLC2A1, a multiple facilitator superfamily (MFS) transporter gene, is particularly susceptible to mutations that cause GLUT1 deficiency syndrome, a human encephalopathy that results from decreased glucose flux through the blood brain barrier. Genotyping of 100 patients revealed that in a third of them who harbor missense mutations in the GLUT1 transporter, transmembrane domain 4 (TM4), encoded by SLC2A1 exon IV, contains mutant residues that have the periodicity of one face of a kinked alpha-helix. Arg-126, located at the amino terminus of TM4, is the locus for most of the mutations followed by other arginine and glycine residues located elsewhere in the transporter but conserved among MFS proteins. The Arg-126 mutants were constructed and assayed for protein expression, targeting, and transport capacity in Xenopus oocytes. The role of charge at position 126, as well as its accessibility, was investigated in R126H by determining its activity as a function of extracellular pH. The results indicate that intracellular charges at the MFS TM2-3 and TM8-9 signature loops and flanking TMs 3, 5, and 6 are critical for the structure of GLUT1 as are TM glycines and that TM4, located at the catalytic core of MFS proteins, forms a helix that surfaces into the extracellular solution where another proton facilitates transport. PMID- 18387951 TI - An essential ligand-binding domain in the membrane receptor for retinol-binding protein revealed by large-scale mutagenesis and a human polymorphism. AB - Plasma retinol-binding protein (RBP), the principal carrier of vitamin A in the blood, delivers vitamin A from liver, the site of storage, to distant organs that need vitamin A, such as the eye, brain, placenta, and testis. STRA6 is a high affinity membrane receptor for RBP and mediates vitamin A uptake in these target organs. STRA6 is a 74-kDa multi-transmembrane domain protein that represents a new class of membrane transport protein. In this study, we used an unbiased strategy by analyzing >900 random mutants of STRA6 to study its structure and function, and we identified an essential RBP-binding domain in STRA6. Mutations in any of the three essential residues in this domain can almost completely abolish binding of STRA6 to RBP and its vitamin A uptake activity from holo-RBP without affecting its cell surface expression. We have also functionally characterized the mutations in human STRA6 that cause severe birth defects as well as several human polymorphisms. All STRA6 mutants associated with severe birth defects have largely abolished vitamin A uptake activity, consistent with the severe clinical phenotypes. In addition, we have identified a human polymorphism that significantly reduces the vitamin A uptake activity of STRA6. Interestingly, the residue affected by this polymorphism is located in the RBP binding domain we identified, and the polymorphism causes decreased vitamin A uptake by reducing RBP binding. This study identifies an essential functional domain in STRA6 and a human polymorphism in this domain that leads to reduced vitamin A uptake activity. PMID- 18387952 TI - Structure-based functional study reveals multiple roles of transmembrane segment IX and loop VIII-IX in NhaA Na+/H+ antiporter of Escherichia coli at physiological pH. AB - The three-dimensional crystal structure of Escherichia coli NhaA determined at pH 4 provided the first structural insights into the mechanism of antiport and pH regulation of a Na(+)/H(+) antiporter. However, because NhaA is activated at physiological pH (pH 6.5-8.5), many questions pertaining to the active state of NhaA have remained open including the structural and physiological roles of helix IX and its loop VIII-IX. Here we studied this NhaA segment (Glu(241)-Phe(267)) by structure-based biochemical approaches at physiological pH. Cysteine-scanning mutagenesis identified new mutations affecting the pH dependence of NhaA, suggesting their contribution to the "pH sensor." Furthermore mutation F267C reduced the H(+)/Na(+) stoichiometry of the antiporter, and F267C/F344C inactivated the antiporter activity. Tests of accessibility to [2 (trimethylammonium)ethyl]methanethiosulfonate bromide, a membrane-impermeant positively charged SH reagent with a width similar to the diameter of hydrated Na(+), suggested that at physiological pH the cytoplasmic cation funnel is more accessible than at acidic pH. Assaying intermolecular cross-linking in situ between single Cys replacement mutants uncovered the NhaA dimer interface at the cytoplasmic side of the membrane; between Leu(255) and the cytoplasm, many Cys replacements cross-link with various cross-linkers spanning different distances (10-18 A) implying a flexible interface. L255C formed intermolecular S-S bonds, cross-linked only with a 5-A cross-linker, and when chemically modified caused an alkaline shift of 1 pH unit in the pH dependence of NhaA and a 6-fold increase in the apparent K(m) for Na(+) of the exchange activity suggesting a rigid point in the dimer interface critical for NhaA activity and pH regulation. PMID- 18387953 TI - The essential function of Tim12 in vivo is ensured by the assembly interactions of its C-terminal domain. AB - The small Tims chaperone hydrophobic precursors across the mitochondrial intermembrane space. Tim9 and Tim10 form the soluble TIM10 complex that binds precursors exiting from the outer membrane. Tim12 functions downstream, as the only small Tim peripherally attached on the inner membrane. We show that Tim12 has an intrinsic affinity for inner mitochondrial membrane lipids, in contrast to the other small Tims. We find that the C-terminal end of Tim12 is essential in vivo. Its deletion crucially abolishes assembly of Tim12 in complexes with the other Tims. The N-terminal end contains targeting information and also mediates direct binding of Tim12 to the transmembrane segments of the carrier substrates. These results provide a molecular basis for the concept that the essential role of Tim12 relies on its unique assembly properties that allow this subunit to bridge the soluble and membrane-embedded translocases in the carrier import pathway. PMID- 18387954 TI - PTP1B regulates cortactin tyrosine phosphorylation by targeting Tyr446. AB - The emergence of protein-tyrosine phosphatase 1B (PTP1B) as a potential drug target for treatment of diabetes, obesity, and cancer underlies the importance of understanding its full range of cellular functions. Here, we have identified cortactin, a central regulator of actin cytoskeletal dynamics, as a substrate of PTP1B. A trapping mutant of PTP1B binds cortactin at the phosphorylation site Tyr(446), the regulation and function of which have not previously been characterized. We show that phosphorylation of cortactin Tyr(446) is induced by hyperosmolarity and potentiates apoptotic signaling during prolonged hyperosmotic stress. This study advances the importance of Tyr(446) in the regulation of cortactin and provides a potential mechanism to explain the effects of PTP1B on processes including cell adhesion, migration, and tumorigenesis. PMID- 18387955 TI - Gamma-aminobutyric acid (GABA) and pentobarbital induce different conformational rearrangements in the GABA A receptor alpha1 and beta2 pre-M1 regions. AB - Gamma-aminobutyric acid (GABA) binding to GABA(A) receptors (GABA(A)Rs) triggers conformational movements in the alpha(1) and beta(2) pre-M1 regions that are associated with channel gating. At high concentrations, the barbiturate pentobarbital opens GABA(A)R channels with similar conductances as GABA, suggesting that their open state structures are alike. Little, however, is known about the structural rearrangements induced by barbiturates. Here, we examined whether pentobarbital activation triggers movements in the GABA(A)R pre-M1 regions. Alpha(1)beta(2) GABA(A)Rs containing cysteine substitutions in the pre M1 alpha(1) (K219C, K221C) and beta(2) (K213C, K215C) subunits were expressed in Xenopus oocytes and analyzed using two-electrode voltage clamp. The cysteine substitutions had little to no effect on GABA and pentobarbital EC(50) values. Tethering chemically diverse thiol-reactive methanethiosulfonate reagents onto alpha(1)K219C and alpha(1)K221C affected GABA- and pentobarbital-activated currents differently, suggesting that the pre-M1 structural elements important for GABA and pentobarbital current activation are distinct. Moreover, pentobarbital altered the rates of cysteine modification by methanethiosulfonate reagents differently than GABA. For alpha(1)K221Cbeta(2) receptors, pentobarbital decreased the rate of cysteine modification whereas GABA had no effect. For alpha(1)beta(2)K215C receptors, pentobarbital had no effect whereas GABA increased the modification rate. The competitive GABA antagonist SR-95531 and a low, non-activating concentration of pentobarbital did not alter their modification rates, suggesting that the GABA- and pentobarbital-mediated changes in rates reflect gating movements. Overall, the data indicate that the pre-M1 region is involved in both GABA- and pentobarbital-mediated gating transitions. Pentobarbital, however, triggers different movements in this region than GABA, suggesting their activation mechanisms differ. PMID- 18387956 TI - Regulation of Gic2 localization and function by phosphatidylinositol 4,5 bisphosphate during the establishment of cell polarity in budding yeast. AB - Establishment of cell polarity is important for a wide range of biological processes, from asymmetric cell growth in budding yeast to neurite formation in neurons. In the yeast Saccharomyces cerevisiae, the small GTPase Cdc42 controls polarized actin organization and exocytosis toward the bud. Gic2, a Cdc42 effector, is targeted to the bud tip and plays an important role in early bud formation. The GTP-bound Cdc42 interacts with Gic2 through the Cdc42/Rac interactive binding domain located at the N terminus of Gic2 and activates Gic2 during bud emergence. Here we identify a polybasic region in Gic2 adjacent to the Cdc42/Rac interactive binding domain that directly interacts with phosphatidylinositol 4,5-bisphosphate in the plasma membrane. We demonstrate that this interaction is necessary for the polarized localization of Gic2 to the bud tip and is important for the function of Gic2 in cell polarization. We propose that phosphatidylinositol 4,5-bisphosphate and Cdc42 act in concert to regulate polarized localization and function of Gic2 during polarized cell growth in the budding yeast. PMID- 18387957 TI - Differential modulation of Akt/glycogen synthase kinase-3beta pathway regulates apoptotic and cytoprotective signaling responses. AB - We have previously reported that specific dopamine agonists mediate protection against apoptosis induced by oxidative stress by activating the D2 receptor coupled phosphoinositide 3-kinase (PI-3K)/Akt pathway. In the present study we examined the downstream effectors of PI-3K/Akt signaling and their role in cell death after oxidative stress and protection provided by ropinirole, a D2 receptor agonist in PC12 cells and primary cultures of dopamine neurons. Ropinirole treatment was associated with rapid translocation and phosphorylation of the PI 3K substrate Akt and phosphorylation of Akt substrates. One of these Akt downstream substrates was identified as the pro-apoptotic factor glycogen synthase kinase-3beta (GSK-3beta). Ropinirole-induced protection was associated with phosphorylation of GSK-3beta (inactivation). In contrast, inhibition of PI 3K blocked the phosphorylation of Akt and GSK-3beta (activation) and prevented the protection mediated by ropinirole. Suppression of Akt with specific short hairpin RNA in normal PC12 cells caused cell death, which was associated with reduced phosphorylation of GSK-3beta and reduced levels of beta-catenin, a transcriptional activator that is regulated by GSK-3beta. Knock-out of GSK-3beta expression with a short hairpin RNA alone was itself sufficient to cause cell death. We further demonstrated that oxidative stress induced by hydrogen peroxide (H2O2) dephosphorylates Akt and GSK-3beta, increases GSK-3beta activity, and promotes an interaction with beta-catenin and its degradation. Inhibition of GSK 3beta activity by inhibitor VIII protects cells from H2O2 similar to ropinirole. These results indicate that GSK-3beta downstream of Akt plays a critical role in cell death and survival in these models. PMID- 18387958 TI - The hyaluronan receptor for endocytosis mediates hyaluronan-dependent signal transduction via extracellular signal-regulated kinases. AB - The hyaluronan (HA) receptor for endocytosis (HARE) mediates the endocytotic clearance of HA and other glycosaminoglycans from lymph and blood. Two isoforms of human HARE, 315- and 190-kDa, are highly expressed in sinusoidal endothelial cells of liver, lymph node, and spleen; HARE is also in specialized cells in the eye, heart, brain, and kidney. Here we determined whether HA binding to HARE initiates intracellular signaling in Flp-In 293 cells stably expressing either the 315- and 190-kDa HARE or the 190-kDa HARE alone. HARE was co immunoprecipitated with extracellular signal-regulated kinase 1 and 2 (ERK1/2), c Jun N-terminal protein kinase (JNK), and p38 members of the mitogen-activated protein kinase signaling cascade. ERK phosphorylation increased in a dose- and time-dependent manner when HA was added to cells expressing full-length or 190 kDa HARE, but not cells with vector-only or a HARE(DeltaLink) construct with greatly decreased ( approximately 90%) HA uptake. HA did not induce phosphorylation of JNK or p38. A maximum increase in phospho-ERK1/2 occurred within 30 min at 5 mug/ml HA, and the response was dampened at >20 mug/ml HA. HA binding did not increase the level of HARE-ERK complexes, but did increase HARE phosphorylation. These findings demonstrate a novel functional response, when HARE binds HA, that leads to activation of ERK1/2, important mediators of intracellular signal transduction. PMID- 18387959 TI - Structural characterization of the primary O-antigenic polysaccharide of the Rhizobium leguminosarum 3841 lipopolysaccharide and identification of a new 3 acetimidoylamino-3-deoxyhexuronic acid glycosyl component: a unique O-methylated glycan of uniform size, containing 6-deoxy-3-O-methyl-D-talose, n acetylquinovosamine, and rhizoaminuronic acid (3-acetimidoylamino-3-deoxy-D-gluco hexuronic acid). AB - Rhizobium are Gram-negative bacteria that survive intracellularly, within host membrane-derived plant cell compartments called symbiosomes. Within the symbiosomes the bacteria differentiate to bacteroids, the active form that carries out nitrogen fixation. The progression from free-living bacteria to bacteroid is characterized by physiological and morphological changes at the bacterial surface, a phase shift with an altered array of cell surface glycoconjugates. Lipopolysaccharides undergo structural changes upon differentiation from the free living to the bacteroid (intracellular) form. The array of carbohydrate structures carried on lipopolysaccharides confer resistance to plant defense mechanisms and may serve as signals that trigger the plant to allow the infection to proceed. We have determined the structure of the major O polysaccharide (OPS) isolated from free living Rhizobium leguminosarum 3841, a symbiont of Pisum sativum, using chemical methods, mass spectrometry, and NMR spectroscopy analysis. The OPS is composed of several unusual glycosyl residues, including 6-deoxy-3-O-methyl-d-talose and 2-acetamido-2deoxy-l-quinovosamine. In addition, a new glycosyl residue, 3-acetimidoylamino-3-deoxy-d-gluco-hexuronic acid was identified and characterized, a novel hexosaminuronic acid that does not have an amino group at the 2-position. The OPS is composed of three to four tetrasaccharide repeating units of -->4)-beta-dGlcp3NAmA-(1-->4)-[2-O-Ac-3-O-Me alpha-d-6dTalp-(1-->3)]-alpha-l-Fucp-(1-->3)-alpha-l-QuipNAc-(1-->. The unique 3 amino hexuronate residue, rhizoaminuronic acid, is an attractive candidate for selective inhibition of OPS synthesis. PMID- 18387960 TI - The impact of a decline in fecundity and of pregnancy postponement on final number of children and demand for assisted reproduction technology. AB - BACKGROUND: Over the past decades, the proportion of couples who resort to infertility treatment has tremendously increased, and fertility (the final number of children) sharply declined. We explored the roles of two potential causes of these trends: a temporal decline in the couples' fecundability and a postponement of age at initiation of pregnancy attempts. METHODS: We conducted a Monte-Carlo simulation for the reproductive history of 100,000 women based on the fertility and socio-demographic characteristics of the 1968 birth cohort in France. Declines in fecundability of various amplitudes have been implemented, as well as increases in the distribution of age at initiation of pregnancy attempts. RESULTS: A decline in fecundability by 15% implied a decrease in fertility by 4%, and an increase in the proportion of couples eligible for infertility treatments by 73%. An increase in the mean age at initiation of first pregnancy attempt by 2.5 years from 25 years entailed a decrease by 5% in fertility and an increase by 32% in the proportion of couples eligible for infertility treatments. CONCLUSION: A relatively important decrease in fecundability and an increase by 2.5 years in age at first pregnancy attempt are likely to have only a limited impact on fertility. However, they may have a large impact on the proportion of involuntarily infertile couples, likely to resort to assisted reproduction techniques. PMID- 18387961 TI - Relevance of anti-Mullerian hormone measurement in a routine IVF program. AB - BACKGROUND: Diminished ovarian reserve has become a major cause of infertility. Anti-Mullerian hormone (AMH) seems to be a promising candidate to assess ovarian reserve and predict the response to controlled ovarian hyperstimulation (COH). This prospective study was conducted to evaluate the relevance of AMH in a routine IVF program. METHODS: Three hundred and sixteen patients were prospectively enrolled to enter their first IVF/ICSI-cycle. Age, FSH-, inhibin B- and AMH-levels and their predictive values for ovarian response and clinical pregnancy rate were compared by discriminant analyses. RESULTS: A total of 132 oocyte retrievals were performed. A calculated cut-off level < or =1.26 ng/ml AMH alone detected poor responders (< or =4 oocytes) with a sensitivity of 97%, and there was a 98% correct prediction of normal response in COH if levels were above this threshold. With levels <0.5 ng/ml, a correct prediction of very poor response (< or =2 oocytes) was possible in 88% of cases. Levels of AMH > or =0.5 ng/ml were not significantly correlated with clinical pregnancy rates. CONCLUSIONS: AMH is a predictor of ovarian response and suitable for screening. Levels < or =1.26 ng/ml are highly predictive of reduced ovarian reserve and should be confirmed by a second line antral follicle count. Measurement of AMH supports clinical decisions, but alone it is not a suitable predictor of IVF success. PMID- 18387962 TI - Congenital anomalies in twins: a register-based study. AB - BACKGROUND: The risk of congenital anomalies in twins is higher than in singletons, but it is less well reported in relation to chorionicity. The aim of this study was to describe the prevalence of congenital anomalies in twin pregnancies by chorionicity and by major subtype and compare the rates with those in singletons. METHODS: The study population included 2329 twin pregnancies (4658 twins) and 147,655 singletons delivered in the Northeast of England during 1998 2002. Data were obtained from the population-based Northern Multiple Pregnancy Register and Northern Congenital Abnormality Survey. RESULTS: The rate of congenital anomalies in twins was 405.8 per 10,000 twins versus 238.2 per 10,000 singletons [rate ratios (RR) = 1.7, 95% confidence interval (CI) 1.5-2.0]. In twins with known chorionicity (84.8% of all twins), the prevalence of congenital anomalies in monochorionic (MC) twins (633.6 per 10,000) was nearly twice that in dichorionic (343.7 per 10,000; RR = 1.8, 95% CI 1.3-2.5). There was an increased rate of congenital anomalies in twin compared with singleton pregnancies for all major types of anomalies, except chromosomal abnormalities. CONCLUSIONS: This study using high quality, population-based data on multiple pregnancies and congenital anomalies found that twins, particularly MC twins, have a higher risk of congenital anomalies than singletons. PMID- 18387963 TI - A systematic review of randomized controlled trials examining psychological interventions for needle-related procedural pain and distress in children and adolescents: an abbreviated cochrane review. AB - OBJECTIVE: To report the results of a systematic review of randomized controlled trials (RCTs) of psychological interventions for children and adolescents undergoing needle-related procedures. METHODS: A variety of cognitive-behavioral psychological interventions for managing procedural pain and distress in children and adolescents between 2 and 19 years of age were examined. Outcome measures included pain and distress as assessed by self-report, observer report, behavioral/observational measures, and physiological correlates. RESULTS: Twenty eight trials met the criteria for inclusion in the review and provided the data necessary for pooling the results. Together, the trials included 1,039 participants in treatment conditions and 951 in control conditions. The largest effect sizes for treatment improvement over control conditions were found for distraction, combined cognitive-behavioral interventions, and hypnosis, with promising but limited evidence for several other psychological interventions. CONCLUSIONS: Recommendations for conducting future RCTs are provided, and particular attention to the quality of trial design and reporting is highlighted. PMID- 18387964 TI - Exon 4 of the human cytomegalovirus (CMV) major immediate-early gene as a target for CMV real-time PCR. PMID- 18387965 TI - Endophthalmitis: potential benefits of repeated intravitreal injections of antibiotics. PMID- 18387968 TI - Functional definition of the mutation cluster region of adenomatous polyposis coli in colorectal tumours. AB - The mutation cluster region (MCR) of adenomatous polyposis coli (APC) is located within the central part of the open reading frame, overlapping with the region encoding the 20 amino acid repeats (20R) that are beta-catenin-binding sites. Each mutation in the MCR leads to the synthesis of a truncated APC product expressed in a colorectal tumour. The MCR extends from the 3' border of the first 20R coding region to approximately the middle of the third 20R coding region, reflecting both positive and negative selections of the N- and C-terminal halves of the APC protein in colon cancer cells, respectively. In contrast, the second 20R escapes selection and can be either included or excluded from the truncated APC products found in colon cancer cells. To specify the functional outcome of the selection of the mutations, we investigated the beta-catenin binding capacity of the first three 20R in N-terminal APC fragments. We found in co immunoprecipitation and intracellular co-localization experiments that the second 20R is lacking any beta-catenin binding activity. Similarly, we also show that the tumour-associated truncations abolish the interaction of beta-catenin with the third 20R. Thus, our data provide a functional definition of the MCR: the APC fragments typical of colon cancer are selected for the presence of a single functional 20R, the first one, and are therefore equivalent relative to beta catenin binding. PMID- 18387969 TI - In tropical lowland rain forests monocots have tougher leaves than dicots, and include a new kind of tough leaf. AB - BACKGROUND AND AIMS: There has been little previous work on the toughness of the laminae of monocots in tropical lowland rain forest (TLRF) despite the potential importance of greater toughness in inhibiting herbivory by invertebrates. Of 15 monocot families with >100 species in TLRF, eight have notably high densities of fibres in the lamina so that high values for toughness are expected. METHODS: In north-eastern Australia punch strength was determined with a penetrometer for both immature leaves (approx. 30 % final area on average) and fully expanded, fully toughened leaves. In Singapore and Panama, fracture toughness was determined with an automated scissors apparatus using fully toughened leaves only. KEY RESULTS: In Australia punch strength was, on average, 7x greater in shade-tolerant monocots than in neighbouring dicots at the immature stage, and 3x greater at the mature stage. In Singapore, shade-tolerant monocots had, on average, 1.3x higher values for fracture toughness than neighbouring dicots. In Panama, both shade-tolerant and gap-demanding monocots were tested; they did not differ in fracture toughness. The monocots had markedly higher values than the dicots whether shade-tolerant or gap-demanding species were considered. CONCLUSIONS: It is predicted that monocots will be found to experience lower rates of herbivory by invertebrates than dicots. The tough monocot leaves include both stiff leaves containing relatively little water at saturation (e.g. palms), and leaves which lack stiffness, are rich in water at saturation and roll readily during dry weather or even in bright sun around midday (e.g. gingers, heliconias and marants). Monocot leaves also show that it is possible for leaves to be notably tough throughout the expansion phase of development, something never recorded for dicots. The need to broaden the botanist's mental picture of a 'tough leaf' is emphasized. PMID- 18387970 TI - Plant growth modelling and applications: the increasing importance of plant architecture in growth models. AB - BACKGROUND: Modelling plant growth allows us to test hypotheses and carry out virtual experiments concerning plant growth processes that could otherwise take years in field conditions. The visualization of growth simulations allows us to see directly and vividly the outcome of a given model and provides us with an instructive tool useful for agronomists and foresters, as well as for teaching. Functional-structural (FS) plant growth models are nowadays particularly important for integrating biological processes with environmental conditions in 3 D virtual plants, and provide the basis for more advanced research in plant sciences. SCOPE: In this viewpoint paper, we ask the following questions. Are we modelling the correct processes that drive plant growth, and is growth driven mostly by sink or source activity? In current models, is the importance of soil resources (nutrients, water, temperature and their interaction with meristematic activity) considered adequately? Do classic models account for architectural adjustment as well as integrating the fundamental principles of development? Whilst answering these questions with the available data in the literature, we put forward the opinion that plant architecture and sink activity must be pushed to the centre of plant growth models. In natural conditions, sinks will more often drive growth than source activity, because sink activity is often controlled by finite soil resources or developmental constraints. PMA06: This viewpoint paper also serves as an introduction to this Special Issue devoted to plant growth modelling, which includes new research covering areas stretching from cell growth to biomechanics. All papers were presented at the Second International Symposium on Plant Growth Modeling, Simulation, Visualization and Applications (PMA06), held in Beijing, China, from 13-17 November, 2006. Although a large number of papers are devoted to FS models of agricultural and forest crop species, physiological and genetic processes have recently been included and point the way to a new direction in plant modelling research. PMID- 18387971 TI - Allocation to reproduction and relative reproductive costs in two species of dioecious Anacardiaceae with contrasting phenology. AB - BACKGROUND AND AIMS: The cost of reproduction in dioecious plants is often female biased. However, several studies have reported no difference in costs of reproduction between the sexes. In this study, the relative reproductive allocation and costs at the shoot and whole-plant levels were examined in woody dioecious Rhus javanica and R. trichocarpa, in order to examine differences between types of phenophase (i.e. physiological stage of development). METHODS: Male and female Rhus javanica and R. trichocarpa were sampled and the reproductive and vegetative allocation of the shoot were estimated by harvesting reproductive current-year shoots during flowering and fruiting. Measurements were made of the number of reproductive and total current-year shoots per whole plant, and of the basal area increment (BAI). The numbers of reproductive and total current-year shoots per 1-year-old shoot were counted in order to examine the costs in the following year at the shoot level. KEY RESULTS: A female-biased annual reproductive allocation was found; however, the ratio of reproductive current-year shoots per tree and the BAI did not differ between sexes in Rhus javanica and R. trichocarpa. The percentage of 1-year-old shoots with at least one reproductive current-year shoot was significantly male-biased in R. trichocarpa, but not in R. javanica, indicating that there was a relative cost at the shoot level only in R. trichocarpa. The female-biased leaf mass per shoot, an indicator of compensation for costs, was only found in R. javanica. CONCLUSIONS: Relative reproductive costs at the shoot level were detected in Rhus trichocarpa, which has simultaneous leafing and flowering, but not in R. javanica, which has leafing followed by flowering. However, the costs for the whole-plant level were diminished in both species. The results suggest that the phenophase type may produce the different costs for R. javanica and R. trichocarpa through the development of a compensation mechanism. PMID- 18387972 TI - Monocot leaves are eaten less than dicot leaves in tropical lowland rain forests: correlations with toughness and leaf presentation. AB - BACKGROUND AND AIMS: In tropical lowland rain forest (TLRF) the leaves of most monocots differ from those of most dicots in two ways that may reduce attack by herbivores. Firstly, they are tougher. Secondly, the immature leaves are tightly folded or rolled until 50-100 % of their final length. It was hypothesized that (a) losses of leaf area to herbivorous invertebrates are generally greatest during leaf expansion and smaller for monocots than for dicots, and (b) where losses after expansion are appreciable any difference between monocots and dicots then is smaller than that found during expansion. METHODS: At six sites on four continents, estimates were made of lamina area loss from the four most recently mature leaves of focal monocots and of the nearest dicot shoot. Measurements of leaf mass per unit area, and the concentrations of water and nitrogen were made for many of the species. In Panama, the losses from monocots (palms) and dicots were also measured after placing fully expanded palm leaflets and whole dicot leaves on trails of leaf-cutter ants. KEY RESULTS: At five of six sites monocots experienced significantly smaller leaf area loss than dicots. The results were not explicable in terms of leaf mass per unit area, or concentrations of water or nitrogen. At only one site was the increase in loss from first to fourth mature leaf significant (also large and the same in monocots and dicots), but the losses sustained during expansion were much smaller in the monocots. In the leaf-cutter ant experiment, losses were much smaller for palms than for dicots. CONCLUSIONS: The relationship between toughness and herbivory is complex; despite the negative findings of some recent authors for dicots we hypothesize that either greater toughness or late folding can protect monocot leaves against herbivorous insects in tropical lowland rain forest, and that the relative importance varies widely with species. The difficulties of establishing unequivocally the roles of leaf toughness and leaf folding or rolling in a given case are discussed. PMID- 18387973 TI - High outcrossing in the annual colonizing species Ambrosia artemisiifolia (Asteraceae). AB - BACKGROUND AND AIMS: Variation in mating patterns may be particularly evident in colonizing species because they commonly experience wide variation in plant density. Here, the role of density for the mating system of Ambrosia artemisiifolia (common ragweed), a wind-pollinated annual colonizing species previously reported as self-compatible, is explored. METHODS: The effect of population density on the proportion of self- and cross-fertilized seeds was examined using allozyme markers and experimental arrays conducted over two seasons in the field. Also the reproductive success of isolated plants located in diverse habitats was measured. The potential occurrence of a physiological mechanism preventing self-fertilization, i.e. self-incompatibility, following controlled self- and cross-pollinations in the glasshouse was examined. KEY RESULTS: Outcrossing rates estimated using allozyme markers were uniformly high, regardless of the spacing between plants. However, when single plants were isolated from congeners they set few seeds. Observations of pollen-tube growth and seed set following controlled pollinations demonstrated that plants of A. artemisiifolia possess a strong self-incompatibility mechanism, contrary to earlier reports and assumptions. CONCLUSIONS: The maintenance of high outcrossing rates in colonizing populations of A. artemisiifolia is likely to be facilitated by the prodigious production of wind-borne pollen, high seed production and extended seed dormancy. PMID- 18387974 TI - HIV risks associated with incarceration among injection drug users: implications for prison-based public health strategies. AB - BACKGROUND: Recent policy announcements in Canada and the United States may potentially affect the risk environment for HIV transmission among incarcerated injection drug users (IDU). We sought to evaluate the potential impact of incarceration on HIV risk behaviour among the IDU enrolled in a prospective cohort study. METHODS: We examined patterns of incarceration among 1247 IDU participants enrolled in a 6-year prospective cohort study in Vancouver, Canada, and tested for potential associations between HIV risk behaviour and incarceration. Correlates of incarceration were identified using generalized estimating equations (GEE). RESULTS: At baseline, factors significantly associated with incarceration included daily injection heroin and injection cocaine use and inconsistent condom use with casual sexual partners. In a GEE analysis, factors independently associated with incarceration included: used syringe borrowing (adjusted odds ratio [AOR] = 1.36; [95% CI: 1.16-1.60]), used syringe lending (AOR = 1.31; [95% CI: 1.12-1.55]) and inconsistent condom use with casual sexual partners (AOR = 1.16; [1.02-1.33]). All variables P < 0.05. CONCLUSION: In our study, incarceration was independently associated with HIV transmission and acquisition behaviours. These findings suggest that increased rates of incarceration of IDU may be associated with increased HIV transmission among this group. PMID- 18387975 TI - The Italian AICE-Genetics hemophilia A database: results and correlation with clinical phenotype. AB - BACKGROUND: The high mutational heterogeneity of hemophilia A is a challenge for the provision of genetic services. We plan to identify the mutation in patients with hemophilia A in order to create a confidential national database of mutations for the optimization of genetic services in Italy. DESIGN AND METHODS: The factor VIII gene (F8) was analyzed in 1296 unrelated patients with hemophilia A using screening methods for intron 22 and 1 inversions and rare mutations (denaturing high performance liquid chromatography, conformation sensitive gel electrophoresis) and/or direct sequencing. RESULTS: F8 mutations were identified in 874 (89%), 146 (89%), and 133 (94%) families with severe, moderate, or mild hemophilia A, respectively. Mutations predicting a null allele were responsible for 80%, 15%, and less than 1% of cases of severe, moderate, or mild hemophilia A, respectively. About 40% of missense and nonsense mutations occurred at a CpG site, arginines being most frequently affected. Of the small deletions or insertions, 29% occurred at one of two stretches of adenines, codons 1191-1194 (8As) and 1439-1441 (9As). Overall, these "hotspots" accounted for 31% of the point mutations in the patients with hemophilia A. Inhibitors developed in 22% of the patients with severe hemophilia A, 8% of those with moderate disease and in 4% of patients with mild hemophilia A. Patients who had severe hemophilia A and mutations predicting a null allele developed inhibitors more frequently (22 to 67%) than patients with missense mutations (5%). CONCLUSIONS: We report a wide spectrum of mutations in a large national database. The type of mutation was a strong predictor of the clinical phenotype. This database is expected to considerably improve the genetic counselling and medical care of families with hemophilia A in Italy. PMID- 18387976 TI - Follow-up of healthy donors receiving granulocyte colony-stimulating factor for peripheral blood progenitor cell mobilization and collection. Results of the Spanish Donor Registry. AB - BACKGROUND: Information about the long-term follow-up and safety of granulocyte colony-stimulating factor administration to healthy donors is limited. The aims of this study were to analyze the side effects of granulocyte colony-stimulating factor administration in donors included in a Spanish Registry of hematopoietic stem cell donors and to determine the long-term outcome of these donors. DESIGN AND METHODS: The Spanish National Donor Registry was developed to record the short- and long-term results of granulocyte colony-stimulating factor administration to mobilize peripheral blood progenitor cells in normal donors. To date, 1436 donors (771 males, 665 females) with a median age of 37 years (range, 1 to 74 years) have been registered. Granulocyte colony-stimulating factor was the only cytokine administered. A baseline investigation was performed in every donor before granulocyte colony-stimulating factor administration and follow-up investigations (controls) were planned at 4 weeks and annually thereafter for up to 5 years after the mobilization. RESULTS: At least one of the scheduled controls was performed in 736 donors, while 320 donors have been followed for 2 years or more. The peripheral white blood cell count decreased significantly from 6.8 x 10(9)/L at baseline to 5.9 x 10(9)/L at 4 weeks after leukapheresis (p<0.0001) and remained at values lower than those observed premobilization until 2 years after mobilization. In contrast, hemoglobin concentration and platelet count returned to normal values within 1 year after mobilization. Bone pain (90%) and headache (33%) were the most frequently reported granulocyte colony stimulating factor-related side effects. Five patients (0.68%) were diagnosed as having solid tumors (lung cancer in two patients and thyroid carcinoma, choroid melanoma, and colon carcinoma in one patient each) between 10 and 64 months after administration of granulocyte colony-stimulating factor. No hematologic malignancies have been reported. CONCLUSIONS: The clinical side effects of granulocyte colony-stimulating factor administration in healthy donors are generally mild. Changes in blood counts were minimal and mainly affected white blood cell counts, which returned to normal values within 2 years after granulocyte-colony stimulating factor administration. No patient developed a hematologic malignancy. A larger number of donors and longer follow-up are needed to determine the safety of granulocyte colony-stimulating factor administration definitively. PMID- 18387977 TI - Selective influences in the expressed immunoglobulin heavy and light chain gene repertoire in hairy cell leukemia. AB - BACKGROUND: We previously reported ongoing mutational and isotype switch events in the immunoglobulin (Ig) heavy chain (H) locus in hairy cell leukemia. Those analyses raised questions on the incidence and type of selective influences occurring on the tumor B-cell receptor of hairy cell leukemia. DESIGN AND METHODS: To further investigate this issue, we examined the full IGH and kappa and lambda light chains (IGkappa and IGlambda) variable and constant region transcripts expressed in a large cohort of patients with hairy cell leukemia (n=88). RESULTS: Multiple IgH isotypes were expressed in 46/56 (82%) cases of hairy cell leukemia. Comparison of tumor with normal B-cell repertoires revealed preferential usage of IGHV3-21, IGHV3-30 and IGHV3-33 in hairy cell leukemia (p=0.001, p=0.003 and p=0.001, respectively). Light chain analysis demonstrated preferential Igl use with an inverted IGk:IGl ratio (0.7:1) and universal usage of IGLJ3. Analysis of LCDR3 junctions revealed highly homologous motifs in 40% of IGL. Parallel analysis of IGH and IGL showed selective pairing of IGHV3-21/30/33 segments to specific LCDR3-J3 subsets (p=0.008). Of 40 cases of hairy cell leukemia, 38 had mutated IGHV and/or IGK/LV, with variations in 13/13 cloned cases, while two had 100% unmutated IGHV and IGK/LV. CONCLUSIONS: Overall, biased IGV usage, preference for Iglambda with universal IGLJ3 usage and a high incidence of LCDR3 homologous motifs suggest selective influences on the B-cell receptor of hairy cell leukemia. Ongoing mutations and isotype switching suggest that influences occur on the tumor B-cell receptor at ectopic sites. PMID- 18387979 TI - Simultaneous genotyping of coagulation factor XI type II and type III mutations by multiplex real-time polymerase chain reaction to determine their prevalence in healthy and factor XI-deficient Italians. AB - BACKGROUND: Factor XI deficiency is a rare autosomal recessive coagulopathy, which is, however, common among Ashkenazi Jews, in whom the so-called type II (E117X) and type III (F283L) mutations account for 98% of alleles. In non-Jewish populations, a higher level of allelic heterogeneity has been reported. However, the type II mutation was found in individuals from England, Portugal, and Italy, and haplotype analysis confirmed its Jewish origin. The aims of this study were to develop a rapid and accurate assay for the simultaneous detection of type II/type III mutations and to determine the frequency of these mutations in a large Italian population of healthy individuals and in a cohort of factor XI deficient Italian patients. DESIGN AND METHODS: Type II and III mutations were detected using a newly developed multiplex four-color real-time polymerase chain reaction assay. Haplotype analysis was performed by either DNA sequencing or fragment-length analysis. RESULTS: Both type II and type III mutations were found among 3879 healthy Italians with an allele frequency of 0.00064 and 0.00051, respectively. Among the 31 analyzed factor XI-deficient patients, the type II mutation was found in three individuals in the homozygous state and in eight individuals in the heterozygous state (one compound heterozygote type II/III). Haplotype analysis revealed the Jewish origin of both mutations. CONCLUSIONS: The newly developed assay is highly specific and reliable (0.02% false positives); and offers a useful means for the molecular diagnosis of factor XI deficiency. Type II and III mutations are present in the Italian population and should be searched for first in factor XI-deficient patients. PMID- 18387978 TI - AB0 blood group and risk of venous or arterial thrombosis in carriers of factor V Leiden or prothrombin G20210A polymorphisms. AB - BACKGROUND: Routine analyses for thrombophilia include determination of the presence of factor V Leiden and prothrombin 20210A polymorphisms. However, the usefulness of these determinations is controversial and the clinical benefit remains questioned because of the moderate risk of associated thrombosis in carriers. In the search for clusters of thrombotic risk factors to estimate individual risk better, we studied the effect of AB0 blood group, a highly prevalent factor with mild prothrombotic features, on the risk and severity of venous and arterial thromboses in carriers of these polymorphisms. DESIGN AND METHODS: We genotyped the AB0 blood group in 981 carriers of factor V Leiden or prothrombin 20210A polymorphisms. In order to avoid the over-representation of a particular genotype and to suppress confounding factors, we included only non related heterozygous carriers without additional genetic risk factors. We studied 609 patients with venous thromboembolism (287 with factor V Leiden, and 322 with prothrombin 20210A), 174 patients with myocardial infarction (78 with factor V Leiden, and 96 with prothrombin 20210A), and 198 controls (96 with factor V Leiden, and 102 with prothrombin 20210A). RESULTS: Non-OO blood group did not increase the risk of myocardial infarction in carriers of factor V Leiden or prothrombin 20210A. However, non-OO blood group contributed significantly to the expression of venous thrombosis associated with both factor V Leiden (OR: 1.76; 95%CI: 1.06-2.91) and prothrombin 20210A (OR: 2.17; 95%CI: 1.33-3.53). Exclusion of A(2)A(2) and A(2)O from the non-00 blood group (because factor VIII-von Willebrand factor levels are similar in these and the 00 blood group) increased the thrombotic risk. Finally, non-OO blood group was associated with an earlier onset in symptomatic carriers of these polymorphisms. CONCLUSIONS: Our study suggests that non-OO blood group increases the risk and severity of venous thrombosis in carriers of prothrombotic polymorphisms. Thus, AB0 phenotyping or genotyping analyses may be valuable components in assessing future thrombophilic risk profiles and might have implications for the policy of thrombosis prophylaxis and treatment. PMID- 18387980 TI - Near infrared spectroscopy (NIRS) in children. AB - Near infrared spectroscopy (NIRS) is a noninvasive method for the in vivo monitoring of tissue oxygenation. Originally used predominantly to assess cerebral oxygenation, NIRS has gained widespread popularity in many clinical settings in all age groups. Changes in regional tissue oxygenation as detected by NIRS may reflect the delicate balance between oxygen delivery and consumption in more than one organ system. However, more studies are required to establish the ability of NIRS monitoring to improve patient outcome. This review provides a comprehensive description of NIRS in children. PMID- 18387981 TI - Regional anesthesia in elderly patients undergoing carotid surgery: report of a case series. AB - BACKGROUND: This study evaluates the short-term results in patients more than 75 years of age undergoing carotid endarterectomy at a single institution. METHODS: Between June 2004 and June 2007, carotid endarterectomy operations were performed in 123 patients. A total of 70 patients had regional anesthesia. The data for all patients were retrospectively reviewed. Regional anesthesia and selective shunting was performed in all patients. RESULTS: In 6 patients, a shunt was required. Primary closure of the carotid artery was performed in 22 patients and patch angioplasty was used in the remainder. There were no postoperative neurological complications. One patient died due to myocardial infarction. CONCLUSIONS: Carotid endarterectomy with regional anesthesia can be performed safely in the elderly population with low mortality and morbidity. Regional anesthesia may have advantages over general anesthesia and could potentially aid in avoiding complications related to shunt use. PMID- 18387982 TI - Prevalence of thrombophilic mutations and ACE I/D polymorphism in Turkish ischemic stroke patients. AB - The aim of this study was to evaluate the prevalence of factor V Leiden (FVL), prothrombin G20210A, methylenetetrahydrofolate reductase (MTHFR) C677T gene mutations, and angiotensin-converting enzyme (ACE) I/D polymorphism in ischemic stroke (IS) patients. A total of 162 Turkish IS patients were included and analyzed according to stroke subtype by the TOAST classification. Their genotype data were compared with those of the control group, representing the healthy population, using the chi(2) test. The frequency of FVL heterozygocity was 12.3% in this series-higher than that in the normal population (9.8%; statistically insignificant, P = .478). The frequency of the ACE D/D genotype in all stroke patients and those with stroke of undetermined etiology was higher than that in our population (52.5% and 59.2%, respectively, vs 39.3%; statistically significant, P = .034, P = .020). Our results may suggest that ACE D/D genotype is a risk factor for IS, particularly in those with stroke of undetermined etiology in the Turkish population. PMID- 18387983 TI - Prevalence of platelet dysfunction and abnormal coagulation: results of a population-based study. AB - The prevalence of impairments in the hemostatic process is unknown in acutely ill people. Data on hemostasis (PFA 100) and the coagulation cascade of 1015 people are presented here, establishing a cohort of unselected emergency patients in a population-based approach. A high prevalence of reduced platelet function (38%) was found, which was more frequent than expected. In contrast, there was a lower prevalence (20%) of abnormal plasmatic coagulation, which was almost always explained by medication, whereas medication could not predict abnormal platelet function. Moreover, a history of disproportionate bleeding did not correlate well with abnormal platelet or coagulation factor function and could not substitute for a screening in this setting. The effect of acetylsalicylic acid (ASA) on PFA closure time was frequently missing (34%), indicating a considerable prevalence of ASA nonresponse among the study population. These data should be applicable in similar settings. The high prevalence of unexpectedly abnormal platelet function in acute illness as well as the high prevalence of possible ASA nonresponders suggests a functional platelet assay to be effective in screening certain subpopulations of emergency patients. PMID- 18387984 TI - Myocardial infarction in a 28-year-old thalassemia intermedia patient. AB - A 28-year-old Lebanese thalassemia intermedia (TI) patient with homozygous IVS1 110 mutation sustained atypical chest pain of 1 day's duration. The EKG reading revealed ST segment elevation in the chest leads V(1) to V(5). Coronary angiography showed 2 plaques in the left anterior descending coronary artery. He underwent subsequent angioplasty with stenting of the left anterior descending coronary artery. An extensive thrombophilia profile was negative. He was started on medication, and his medical condition improved and chest pain ceased. This is the first case report of myocardial infarction in a TI patient among thalassemics. We propose that such cases will emerge more frequently as our population ages, keeping in mind a possible thrombotic mechanism. PMID- 18387985 TI - Methodological issues in genetic association studies of inherited thrombophilia: original report of recent practice. AB - The aims of this article are to evaluate the methodological quality of genetic association studies on the inherited thrombophilia published during 2003 to 2005, to identify the most common mistakes made by authors of those studies, and to examine if overall quality of the article correlates with the quality of the journal. Articles were evaluated by 2 independent reviewers using the checklist of 16 items. A total of 58 eligible studies were identified. Average total score was 7.59 +/- 1.96. Total article score did not correlate with the journal impact factor (r = 0.3971; 95% confidence interval [CI], 0.1547-0.5944, P = .002). Total score did not differ across years (P = .624). Finally, it is concluded that methodological quality of genetic association studies is not optimal, and it does not depend on the quality of the journal. Journals should adopt methodological criteria for reporting the genetic association studies, and editors should encourage authors to strictly adhere to those criteria. PMID- 18387986 TI - Macroscopic findings at appendicectomy are unreliable: implications for laparoscopy and malignant conditions of the appendix. AB - BACKGROUND: At laparoscopic appendicectomy, some surgeons leave a macroscopically normal appendix in situ. To assess if this is safe practice, the authors wanted to identify the correlation between macroscopic findings at appendicectomy and pathologists' assessment with regard to the inflamed and the neoplastic. METHOD: Operative cases and histological findings over a 3-year period were identified. RESULTS: The authors obtained 876/972 notes (90.1%). There was 100% congruity between surgeons and pathologists when perforation, abscess, or gangrene of the appendix was noted (n = 235). Where inflammation was the sole recorded finding, 8.3% of cases (37/445) were histologically normal; 33.1% (47/139) of appendices perceived to be normal at appendicectomy demonstrated histological signs of inflammation. Of 16 neoplastic lesions only 3 were identified at the time of surgery. CONCLUSIONS: Surgeons' ability to diagnose a normal appendix is poor as is the ability to detect neoplastic lesions. At laparoscopy, to investigate acute abdominal pain a macroscopically normal appendix should be removed. PMID- 18387987 TI - Parietal cell (oncocytic) adenocarcinoma of the stomach in a female patient: superficial spreading and extensive nodal involvement. AB - A case of parietal cell (oncocytic) adenocarcinoma in a 62-year-old woman is reported. This rare entity has 26 reported cases to date and is usually related to male gender and localized disease with favorable prognosis. In the present case, the unusual finding of parietal cell (oncocytic) adenocarcinoma and extensive nodal metastasis in a woman is reported. PMID- 18387988 TI - Prognostic significance of micropapillary pattern in pure mucinous carcinoma of the breast. AB - Breast carcinoma with micropapillary architecture is associated with aggressive behavior. Similar micropapillary pattern in pure mucinous carcinoma has been noticed and has been shown to convey poor prognosis. In this study 17 cases of pure mucinous carcinoma of the breast seen during a 10-year period have been reviewed, with special reference to micropapillary pattern. Diffuse micropapillary pattern was seen in 6 of 17 cases of mucinous carcinoma of the breast and demonstrated reverse polarity immunostaining pattern with "Epithelial Membrane Antigen." In all cases, the tumor cells showed grade I morphology, and no lymph node metastases were noticed. All the tumors except 1 expressed strong estrogen and progesterone receptor expression, however, all the cases were negative for Her-2/neu expression. In this present study, mucinous carcinomas with micropapillary pattern showed a low nuclear grade, higher incidence of hormone receptor positivity, and lower incidence of Her-2/neu similar to mucinous carcinomas without micropapillary pattern, thus explaining their indolent behavior. PMID- 18387989 TI - Review article: relationship of human papillomavirus with papillary squamous cell carcinoma of the upper aerodigestive tract: a review. AB - The purpose of this review is to evaluate case reports of papillary squamous cell carcinoma (PSCC) of the upper aerodigestive tract (UADT) to assess its relationship with human papillomavirus (HPV). The medical literature was searched for case reports of this condition. A total of 115 cases of PSCC were found that described the condition in sufficient detail. HPV detection was performed in only 22 of the 115 cases of PSCC (19%), and 11 of the 22 cases (50%) are related to this virus. The majority of cases related to HPV are produced by low-risk HPV type 6 followed by high-risk HPV type 16. Today, the association of HPV with PSCCs seems unclear because in the majority of patients tests were not performed for the detection of the HPV. This association should be clearly established to make a correct diagnosis and propose the best therapeutic strategies, such as new vaccines. PMID- 18387990 TI - Isolated tumor cells and circulating CK20 mRNA in pN0 colorectal cancer patients. AB - Micrometastases in lymph nodes and blood may provide important prognostic information. In this study, cytokeratin 20 (CK20) positive cells in lymph nodes and circulating CK20 mRNA were studied using 57 paraffin-embedded lymph node specimens and blood from 24 patients with pN0 colorectal cancer (CRC), respectively. Results showed that 29 out of 56 (52%) lymph node specimens had CK20-positive cells (range: 1-35). Follow-up of the patients for 12 months indicated that 4 patients (7%) had CRC metastases to liver, lung, and bone. In addition, 8 out of 24 (33%) samples had at least 2-fold circulating CK20 mRNA expression higher than the pooled normal sample. This study provides evidence that CK20-positive cells were found in the lymph nodes and differentially expressed circulating CK20 mRNA was also detected in the blood from patients with pN0 CRC. Long-term follow-up is necessary to study their prognostic use in patients with non-metastatic CRC. PMID- 18387991 TI - Granular cell tumor of the parotid gland: an exceptionally rare occurrence. AB - The authors report on a case of a granular cell tumor of the parotid gland, which constitutes an extremely rare neoplasm. The infiltrative growth of the tumor may pose the question of malignancy, especially on frozen section. The objective of this report was to function as a reminder of this entity for every pathologist that deals with a biopsy from a lesion of the parotid gland, and furthermore, to keep the unwary away from a possible diagnostic pitfall. A meticulous search of the literature revealed only 4 similar cases. PMID- 18387992 TI - Primary synovial sarcoma of the prostate: report of 2 cases and literature review. AB - Two cases of primary prostatic synovial sarcoma presenting as a prostatic mass are presented in patients aged 44 and 46 years. Histologically, both tumors were mainly composed of uniform spindle cells forming interlacing fascicles. Clusters of immature epithelioid cells were also observed among the spindle cells in case 1. Immunohistochemically, the tumor cells of both cases were strongly positive for vimentin, bcl-2, CD99, and E-cadherin, as well as focally positive for cytokeratin. However, they were negative for prostate-specific antigen, S-100 protein, CD34, CD117, muscle-specific actin, desmin, and calretinin. The presence of an SYT-SSX gene fusion resulting from t(X;18) was demonstrated from paraffin blocks by reverse transcriptase polymerase chain reaction in both cases. To the authors' knowledge, these represent the fifth and sixth reported cases of prostatic synovial sarcoma. Accurate diagnosis depends on morphologic and immunohistochemical examination and proper molecular analysis. PMID- 18387993 TI - Merkel cell carcinoma arising in the ear canal. AB - A case of rare tumor, Merkel cell carcinoma, located in the ear canal of a 25 year-old woman is presented. A polypoid tumor mass was extirpated, and tympanoplasty was done at the first operation, whereas at the second operation, all the bones of the ear canal were removed. Epitympanum and cavum were filled with tumor, and the tumor mass was removed in toto. The histopathology and immunohistochemical staining characteristics of tumor confirmed the presence of Merkel cell tumor. Postoperatively, radiation therapy to the tumor bed was completed. There was no clinical or radiographic evidence of recurrence or metastasis of Merkel cell tumor for 3 years. PMID- 18387994 TI - CD30-positive T-cell lymphoproliferative disorder of the oral mucosa--an indolent lesion: report of 4 cases. AB - Four cases of CD30-positive T-cell lymphoproliferative disorder (CD30+ LPD) of the oral mucosa are described. This article aims to draw attention to this entity and to emphasize its usual benign clinical behavior despite its resemblance to T cell lymphoma. All the patients were adults. Three of the lesions were on the dorsal surface of the tongue and 1 affected the buccal mucosa. All biopsies showed a dense lymphoid infiltrate composed of CD30+ atypical T cells with a polymorphous infiltrate in the background, which included eosinophils. In 1 case, monoclonal T-cell expansion was detected by molecular techniques. Three cases tested for Epstein-Barr virus were all negative. It is concluded that primary CD30+ T-cell LPD of the oral mucosa can be regarded as the oral counterpart of cutaneous CD30+ LPD such as lymphomatoid papulosis or anaplastic large cell lymphoma. Recognition of the condition is important to avoid overtreatment. PMID- 18387996 TI - The potentially lethal nature of bronchial casts: plastic bronchitis. AB - The gross and microscopic findings from 2 fatal cases of plastic bronchitis (PB) in children, a rare entity characterized by the formation of large mucous casts in the bronchial tree, are presented. These casts differ from ordinary mucus because of their increased cohesiveness and consistency, resulting in solid structures that model the respiratory airway tree. PB usually presents as a complication of underlying diseases, which determine the prognosis of the afflicted patients. Conditions commonly associated with PB include congenital cardiopathies, lymph vessel malformations, asthma, and sickle cell disease. The first case is a typical example of PB following surgical treatment of a cyanotic cardiopathy, with very characteristic and illustrative morphology. The second case describes the fatal course of PB in a patient with sickle cell disease, which has never been reported to the best of the authors' knowledge; in this case, an overlooked association with asthma could have contributed to the fatal outcome. PMID- 18387995 TI - Cystic lesion mimicking intraductal papillary mucinous tumor arising in heterotopic pancreas of the stomach and synchronous intraductal papillary mucinous adenocarcinoma of the pancreas. AB - This article presents the study of a 66-year-old man with an asymptomatic pancreatic mass detected incidentally 4 months earlier. A magnetic resonance imaging scan revealed 2 distinct cystic masses in the pancreas and the gastric antrum. Microscopically, the pancreatic lesion showed dilated cysts containing papillary structures lined by mucinous epithelium, which showed a loss of polarity, an increased nucleus-to-cytoplasm ratio, a prominent nucleolus, and high proliferation on immunostaining for Ki-67. The gastric lesion was composed of heterotopic pancreatic tissue surrounding a large dilated cyst that was lined with mucinous epithelium and contained a few intraluminal papillae. PMID- 18387997 TI - Primary hepatic lymphoma of mucosa-associated lymphoid tissue type: a case report with cytogenetic study. AB - Primary hepatic lymphoma of mucosa-associated lymphoid tissue type is extremely rare. Only 38 cases have been reported to date. A case of a 59-year-old man with Helicobacter pylori-resistant gastric ulcers and Buerger disease who was followed up since 1999 is reported. A 2-cm hepatic nodule was incidentally found during partial gastrectomy and corresponded to mucosa-associated lymphoid tissue-type lymphoma without underlying liver disease. Molecular studies showed a clonal immunoglobulin heavy-chain gene rearrangement. Investigations for the mucosa associated lymphoid tissue lymphoma-associated translocations t(11;18) and t(14;18), as well as the t(3;14)(q27;q32), were negative, whereas trisomy 3 and trisomy 18 were detected. PMID- 18387998 TI - Renal cell carcinoma with sarcomatoid features and peritumoral sarcoid-like granulomatous reaction: report of a case and review of the literature. AB - Granulomatous inflammation with multinucleated giant cells is observed in various infectious and noninfectious diseases. It has been found in association with malignant tumors and designated sarcoid-like reaction. The distinction between a tumor-related granulomatous reaction and a true sarcoidosis can be a problematic issue. A case of renal cell carcinoma with sarcomatoid features (Fuhrman nuclear grade IV) with an extensive peritumoral sarcoid-like reaction and a critical review of the few cases of this association described in the literature have been reported, and the problematic clinical and pathological assessments of such lesions are discussed. PMID- 18387999 TI - Proficiency-based laparoscopic simulator training leads to improved operating room skill that is resistant to decay. AB - The aim of this study was to assess skill retention in the operating room following completion of a proficiency-based laparoscopic skills curriculum. Novices (n = 15) were randomized to a control and a training group that practiced to proficiency on the Fundamentals of Laparoscopic Surgery suturing model. The performance of both groups was assessed on the simulator and on a live porcine laparoscopic Nissen fundoplication model at training completion (posttest) and 5 months later (retention test). Training to proficiency required 4.7 +/-1.2 hours and 41 +/- 10 repetitions. Trained participants outperformed controls, and their performance deteriorated slightly between posttests and retention tests on the simulator (505 +/- 22 vs 462 +/- 50, respectively; P < .05) but not in operating room (263 +/- 138 vs 279 +/- 88, respectively; P = .38). Proficiency-based simulator training results in durable improvement in operative skill of trainees even in the absence of practice for up to 5 months. Minute simulator performance changes do not translate to the operating room. PMID- 18388000 TI - A high-definition exoscope system for neurosurgery and other microsurgical disciplines: preliminary report. AB - An 8-mm diameter rigid lens telescope with a focal distance of 20 cm was developed for open microsurgery. The telescope was attached to a 3-chip high definition digital camera and then to a high-definition monitor. A pneumatic scope holder permitted repositioning. The optical quality of the device was compared with the operating microscope with a step wedge and 1-mm grid paper. Craniotomies and microsurgical dissections with the telescope system (high definition exoscope system) were performed in a live pig model. The high definition exoscope system provided image quality that rivaled the operating microscope even at high magnification. The system was easy to manipulate and comfortable during neurosurgical operations. The lack of stereopsis was a relative drawback of the system but was compensated for with repeated procedures. Overall, this prototype telescope-based system rivals the operating microscope optical quality and field of view. With further refinement, this system could have widespread application in many microsurgical disciplines. PMID- 18388001 TI - Association of demographic and treatment variables in long-term colon cancer survival. AB - The purpose of this study is to examine demographic and treatment variables because they relate to 5-year survival in colon cancer. The study design is analysis of 174 471 patients with colon and rectosigmoid cancer as reported to the American College of Surgeons National Cancer Data Base. Factors associated with a reduced risk of mortality included female gender (hazard ratio = 0.89; 95% confidence interval, 0.87-0.90), education status (hazard ratio = 0.87; 95% confidence interval, 0.85-0.89), increased number of lymph nodes resected (compared with <8, 8-12: hazard ratio = 0.90; 95% confidence interval, 0.89-0.92; >12: hazard ratio = 0.79; 95% confidence interval, 0.77-0.80), and addition of chemotherapy (hazard ratio = 0.69; 95% CI, 0.68-0.71). African American race (hazard ratio = 1.14; 95% confidence interval, 1.11-1.18) and increasing age correlated with an increased hazard risk (61-75 years: hazard ratio = 1.26; 95% confidence interval, 1.23-1.29; >or=76 years: hazard ratio = 2.15; 95% confidence interval, 2.09-2.21, compared with age <60 years). Survival in colon cancer is significantly impacted by patient's age, race, gender, and education status but not by income or area of residence. PMID- 18388002 TI - Body esteem improves after bariatric surgery. AB - Body esteem is an issue for the morbidly obese. Although the primary goal of bariatric surgery is to improve, cure, and prevent medical comorbidities, the psychological aspect of bariatric surgery is just as important. Few studies have investigated the body esteem of patients after laparoscopic gastric bypass. This investigation tested the hypothesis that body esteem improves after bariatric surgery. Preoperative and postoperative patients were asked to fill out an institutional review board-exempted survey that included the Body-Esteem Scale for Adolescents and Adults (BESAA). The subscales include Appearance, Weight, and Attribution. Postoperative patients were told to fill the BESAA as they felt currently and as they felt before surgery. They felt that they had better scores currently than before surgery. Preoperative patients had worse scores than postoperative patients. As in many medical issues, body esteem improves after bariatric surgery. When discussing its benefits, psychological aspects of body esteem should be touted as well. PMID- 18388003 TI - An unlikely pioneer in laparoscopy: Benjamin Henry Orndoff, MD. PMID- 18388004 TI - Commentary on "Laser and radiofrequency endovenous ablation of venous reflux". PMID- 18388005 TI - Commentary on "Carotid artery stenting in the SPACE and EVA-3S trials: analysis and update". PMID- 18388006 TI - Commentary on "Sclerotherapy: a truly minimally invasive technique". PMID- 18388007 TI - Commentary on "Limb ischemia: medical therapy trial update". PMID- 18388008 TI - Laser and radiofrequency endovenous ablation of venous reflux. AB - Endovenous modalities to treat superficial venous reflux of the lower extremities have revolutionized management of patients with varicose veins. Laser and radiofrequency probes have both found their way into the arsenal of physicians treating venous reflux. Although both offer distinct advantages and minor drawbacks, they each offer the convenience of in-office treatment, faster recovery, and improved safety over traditional surgical procedures. This article will briefly discuss the technique, treatment results, and potential complications associated with each procedure. PMID- 18388009 TI - Endovascular management of deep venous thrombosis. AB - The past decade has witnessed new developments for the treatment of deep venous thrombosis (DVT) as well as more information about the virulent nature of DVT over the long term. Symptoms of pain, edema, skin changes, and/or ulceration can affect upwards of 70% of individuals to some degree. Studies have determined that early intervention of thrombus removal may help prevent postthrombotic syndrome in a significant number of patients. Several devices now specifically combine mechanical or ultrasound energy with chemical thrombolysis. These devices include the Trellis-8, Angiojet Power Pulse System, and the Ekos Endowave. Other important aspects central to successful endovascular removal of DVT include using ultrasound guidance for access, understanding venous anatomy and physiology in relation to endovascular techniques, knowing when to perform venous interventions, and using intravascular ultrasound. Endovascular removal of DVT is increasingly becoming the standard of care, particularly that affecting the iliofemoral segments. PMID- 18388010 TI - Commentary on "Kissing stent reconstruction of the aortoiliac bifurcation". PMID- 18388011 TI - Commentary on "Embolic protection in infrainguinal interventions". PMID- 18388012 TI - Kissing stent reconstruction of the aortoiliac bifurcation. AB - Kissing stent reconstruction of the aortoiliac bifurcation is a widely used technique for the management of aortoiliac occlusive disease involving the aortic bifurcation or proximal common iliac arteries. New advances in delivery systems and stent design have enabled better anatomic results with kissing stenting. Long term patency is generally excellent, although several factors may adversely affect patency and should be taken into account when devising the stenting configuration and selecting the device to be used. Geometric variables related to individual aortic anatomy and disease pattern (patient dependent) and stenting configuration (operator dependent) may have an impact on long-term patency. Kissing stent aortoiliac reconstruction is effective and durable, even with complex aortoiliac disease and long-segment occlusions. In our experience, TASC (TransAtlantic Inter-Society Consensus) C and D class disease is not necessarily a contraindication to aortoiliac stent reconstruction, especially in poor operative risk patients. Most restenoses are amenable to endovascular treatment, with excellent long-term assisted patency. PMID- 18388013 TI - Diagnosis and management of venous ulcers. AB - Venous ulceration of the lower extremities is a common and often disabling condition. Venous ulcers are the result of a chronic inflammatory condition caused by persistent venous hypertension. Therapy is directed at counteracting the chronic inflammation in the tissues and at decreasing ambulatory venous hypertension in the area. Compression therapy helps decrease the venous hypertension and aids healing. Topical agents may be used to help decrease the bacterial load in the wound, provide a moist healing environment for dry wounds, or absorb the exudate in wounds with a lot of drainage. Pharmacological adjuncts, such as pentoxifylline or flavanoids, may help counteract the chronic inflammation in the ulcerated area. Interventions to decrease the ambulatory venous hypertension can help patients with either active or healed ulcers. Ablation of incompetent superficial truncal veins and/or perforating veins using radiofrequency ablation, endovenous laser ablation, or foam sclerotherapy can speed ulcer healing and prevent recurrence. PMID- 18388014 TI - Commentary: Golledge J, et al. The outcome of endovascular repair of small abdominal aortic aneurysms. Ann Surg. 2007; 245:326-333. PMID- 18388015 TI - Commentary: Tsai TT, Fattori R, Trimarchi S, et al. Long-term survival in patients presenting with type B acute aortic dissection: insights from the International Registry of Acute Aortic Dissection. Circulation. 2006;114: 2226 2231. PMID- 18388016 TI - Preventable complications of carotid stenting. AB - With the development of equipment specifically designed for carotid artery stenting (CAS) and a national effort to train specialists, CAS has blossomed into a viable alternative to carotid endarterectomy (CEA) in certain circumstances. Not without risks, stroke is a well-recognized complication of CAS; yet a variety of other unique problems can complicate an expected successful outcome. This review covers some of the more common problems and provides potential solutions to preventable situations that may occur in the periprocedural period. The primary objective of this review is to provide a heightened awareness of both anticipated and unanticipated risks of the procedure, with the hope of avoiding recurring problems. PMID- 18388017 TI - Carotid artery stenting in the SPACE and EVA-3S trials: analysis and update. AB - Carotid artery stenting is an alternative to carotid endarterectomy. Although the stroke and mortality rates after stenting are low, it is yet to be proved that stenting is superior or equal to endarterectomy in low-risk symptomatic and asymptomatic patients. A summary of the results of 2 recent trials comparing carotid stenting with carotid endarterectomy, SPACE, and EVA-3S, is presented herein. Both trials failed to prove noninferiority of carotid stenting compared with carotid endarterectomy. PMID- 18388018 TI - Embolic protection in infrainguinal interventions. AB - The benefits of the use of embolic protection devices (EPDs) in saphenous vein coronary artery bypass grafts and carotid arteries have been shown, but the utility of their application during infrainguinal endovascular interventions is somewhat unclear. Patients with specific anatomical features or lesion characteristics, or patients undergoing specific types of endovascular interventions may benefit from the off-label use of EPDs, but this has yet to be determined. This report will examine the current literature related to the use of EPDs in infrainguinal endovascular interventions and attempt to identify the patients who would most likely benefit from their use. In addition, the setting in which EPDs have been used at one institution is briefly described. This discussion will serve as a general guideline for the use of EPDs and act as an impetus for the development of future clinical trials to help elucidate the patients who will truly benefit from EPDs use when undergoing lower extremity endovascular revascularization procedures. PMID- 18388019 TI - The use of stem cells in the treatment of inoperable limb ischemia. AB - Autologous stem cell-based therapy offers many advantages over other nonsurgical treatments of peripheral arterial disease. Transplant stem cells are a renewable source of multiple cytokines and growth factors. Autologous adult stem cells are not rejected and do not form teratomas. Early preliminary results indicate that adult stem cell therapy is safe and effective in patients with critical limb ischemia. PMID- 18388020 TI - Limb ischemia: medical therapy trial update. AB - Peripheral arterial disease has a significant impact on the quality of a patient's life. Pain, fear of limb loss, increased inactivity, and poor lifestyle choices such as continued smoking further debilitate these patients. Surgical and endovascular procedures have affected this outcome for many but carry the added risk of procedural complications and mortality for this group of high-risk individuals. Risk factor management and medical therapy carry little risk and can also improve functional outcome for many of these patients. Despite this, there are currently only two FDA-approved medications for the treatment of claudication. This review discusses published data on the impact of walking distance with investigational pharmacotherapy. PMID- 18388021 TI - Commentary on "Preventable complications of carotid stenting". PMID- 18388022 TI - Commentary on "Branding of vascular surgery". PMID- 18388023 TI - Branding of vascular surgery. AB - The Society for Vascular Surgery surveyed primary care physicians (PCPs) to understand how PCPs make referral decisions for their patients with peripheral vascular disease. Responses were received from 250 PCPs in 44 states. More than 80% of the respondents characterized their experiences with vascular surgeons as positive or very positive. PCPs perceive that vascular surgeons perform "invasive" procedures and refer patients with the most severe vascular disease to vascular surgeons but were more than twice as likely to refer patients to cardiologists, believing they are better able to perform minimally invasive procedures. Nevertheless, PCPs are receptive to the notion of increasing referrals to vascular surgeons. A successful branding campaign will require considerable education of referring physicians about the totality of traditional vascular and endovascular care increasingly provided by the contemporary vascular surgical practice and will be most effective at the local grassroots level. PMID- 18388024 TI - Commentary: Dubinsky RM, Lai SM. Mortality from combined carotid endarterectomy and coronary artery bypass surgery in the US. Neurology. 2007;68:195-197. AB - available at http://www.neurology.org/cgi/ content/abstract/68/3/195. PMID- 18388025 TI - Antiplatelet therapy for vascular interventions. AB - The role of platelets and the importance of platelet inhibition are well established in patients with vascular disease. When physicians perform revascularization procedures in these patients, who typically have the most advanced atherosclerotic disease, they should be aware of both the generic benefits of platelet inhibition on patients' long-term health as well as the specific benefits relative to operative or endovascular revascularization. Platelet inhibition has not been well studied in the aortoiliac system, renal vascular bed, or the mesenteric arterial bed. Peripheral vascular beds that have been studied are predominantly the carotid circulation and the infrainguinal vasculature. This article reviews the basic mechanisms of platelet inhibition and suggests antiplatelet strategies for vascular interventions, including carotid endarterectomy, carotid angioplasty and stenting, infrainguinal bypass, and peripheral angioplasty and stenting. PMID- 18388026 TI - Commentary on "Diagnosis and management of venous ulcers". PMID- 18388027 TI - Effects of atrial fibrillation on arterial stiffness in patients with hypertension. AB - Arterial stiffness is significantly correlated with cardiovascular risk in patients with hypertension. Although arterial stiffness increases with age and other vascular risk factors, the effect of heart rhythm on arterial stiffness is uncertain. The aim of this study was to determine whether the presence of atrial fibrillation is associated with arterial stiffness, as determined by heart to femoral pulse wave velocity, in patients with hypertension. Heart to femoral pulse wave velocity was measured, and comprehensive transthoracic echocardiography was performed in 68 subjects (35 subjects with sinus rhythm, 33 subjects with atrial fibrillation). Potential determinants of heart to femoral pulse wave velocity were identified by univariate and multivariate analyses. Individuals with atrial fibrillation had higher heart to femoral pulse wave velocity than those with sinus rhythm (1028 +/- 222 vs 923 +/- 110 cm/s, P = .03). Age, the presence of atrial fibrillation, systolic blood pressure, and pulse pressure were found to be significantly correlated with heart to femoral pulse wave velocity. After adjusting for possible con-founders, multivariate analysis identified systolic blood pressure (P = .003) and the presence of atrial fibrillation (P = .007) as independent determinants of heart to femoral pulse wave velocity. The presence of atrial fibrillation was significantly correlated with a higher pulse wave velocity, independently of age or blood pressure in patients with hypertension. PMID- 18388028 TI - Bilateral carotid and bilateral vertebral artery dissection following facial massage. AB - A 50-year-old woman underwent facial massage. After 13 days, she experienced left retro-orbital pain, ptosis, and miosis. Magnetic resonance imaging (MRI) showed stenotic dissection of bilateral cervical internal carotid and vertebral arteries. The intracranial vasculature was intact. She was treated conservatively with long-term oral anticoagulation and remains asymptomatic 18 months later. PMID- 18388029 TI - QT dispersion and left ventricular hypertrophy in elderly hypertensive and normotensive patients. AB - Inhomogeneity of ventricular repolarization as detected by QT dispersion may be a potential leading mechanism of sudden death in hypertensive and normotensive (age related) left ventricular hypertrophy. Aim of this study was to investigate QT dispersion, ventricular arrhythmias, and left ventricular mass index in elderly hypertensive and normotensive patients. Study population consisted of 60 consecutive patients (sex: 34 men/26 women; age: 63 +/- 11 years) with essential arterial hypertension and 48 age and sex-matched control subjects (24 men/24 women; 64 +/- 16 years). Measurements included QTc dispersion, ventricular arrhythmias, and left ventricular hypertrophy. Hypertensive patients had greater left ventricular mass index (P = .006) and higher QTc dispersion (P = .004) than controls. Left ventricular hypertrophy was diagnosed in 57 (31 men/26 women) of all subjects. These patients had higher blood pressure (P < .05), Lown's score (P < .001), and QTc dispersion (P < .001). QTc dispersion and Lown's score were independent predictors of left ventricular mass index (P < .001). Conclusively, QTc dispersion is a strong indicator of left ventricular mass index and might be used in risk stratification of hypertensive and normotensive elderly patients. PMID- 18388030 TI - A retrospective analysis of hospitalized patients with documented deep-venous thrombosis and their risk of pulmonary embolism. AB - The question of when and how intensively patients with deep-venous thrombosis (DVT) can ambulate remains scarcely underlined. The authors evaluated the evolution of DVT by comparing bed rest and mobilization and using all variables potentially relevant to a risk/benefit evaluation in 252 patients. The end points were the progression of the thrombotic disease and the incidence of pulmonary embolism (PE) at 30 days. Immobilized patients had a higher incidence of events (hazard ratio 4.39; P < .0001). At multivariate analysis, immobilization (hazard ratio 2.41; P = .026) and the lack of leg compression (hazard ratio 4.58; P = .009) were the only independent predictors of the clinical end point. This retrospective analysis raises the question as to whether all patients with confirmed DVT should ambulate and receive an adequate leg compression. A prospective study is needed to determine whether immobilization of such patients actually increases their risk of developing pulmonary embolism. PMID- 18388031 TI - Effects of cilostazol and pentoxifylline on forearm reactive hyperemia response, lipid profile, oxidative stress, and inflammatory markers in patients with intermittent claudication. AB - Peripheral arterial disease may lead to lower limb claudication and increased risk of systemic vascular dysfunction. In this article, the authors have investigated the peripheral vascular dysfunction evaluating forearm blood flow using venous occlusion plethysmography, lipid profile, and C-reactive protein in 60 patients with moderate intermittent claudication treated during 20 weeks with placebo (n = 16), cilostazol (200 mg/d; n = 17), or pentoxifylline (1200 mg/d; n = 15) in a randomized double-blinded clinical trial, taking into account smoking. Forearm blood flow after reactive hyperemia response (FBF(h) ) or oral nitroglycerine spray to evaluate endothelial-dependent and endothelial independent vasodilation, respectively, pain-free and maximal walking distance, levels of C-reactive protein, triglycerides, cholesterol, low-density lipoprotein, and high-density lipoprotein-cholesterol in plasma were determined. The results showed that there was an improvement in the high-density lipoprotein cholesterol, pain-free and maximal walking distance, and FBF(h) independent of treatment in nonsmoking patients. Cilostazol increased high-density lipoprotein cholesterol level, maximal walking distance, and FBF(h), whereas pentoxifylline reduced C-reactive protein level and increased maximal walking distance in total and nonsmoking groups. No treatment was effective in smokers. PMID- 18388032 TI - The right atrial thrombus: the sword of Damocles with real risk of massive pulmonary embolism. AB - Cases of 6 patients admitted at the intensive care unit for massive pulmonary embolism are reported. All patients presented with dyspnea, tachypnea, and tachycardia, and 4 were hypotensive and had syncope. Lung ventilation/ perfusion scans revealed perfusion defects in 4 patients. Transthoracic echocardiography (TTE) demonstrated acute cor pulmonale. It also revealed mobile right atrial thrombi in 5 patients, adherent thrombus in the right atrium in 1 patient and patent foramen ovale in 4 patients. Thrombolytic therapy was initiated in 4 patients, and 2 patients received heparin infusion only. Effects of thrombolysis were monitored using bedside TTE during the first 24 hours and in follow-up. The outcome of 4 patients who received thrombolytic therapy was good whereas other 2 patients, who received only heparin, died. Thrombotic mass disappeared 8 to 12 hours after initiation of therapy, and 10 weeks after discharge TTE showed normalized right ventricle dimensions and function in all 4 patients. PMID- 18388033 TI - A-wave acceleration: a new Doppler echocardiographic index for evaluation of left ventricular diastolic dysfunction in elderly patients. AB - Age alters Doppler indexes of left ventricular diastolic performance. Thus, the Doppler detection of left ventricular diastolic dysfunction in the elderly is difficult. The reliability of Doppler indexes in detecting left ventricular diastolic dysfunction in the elderly patients with cardiac diseases known to affect diastolic function were evaluated. Diastolic function using pulsed Doppler in 6 groups of 10 subjects each: elderly normal, young normal, and elderly with hypertrophic cardiomyopathy, aortic stenosis, coronary artery disease, and dilated cardiomyopathy was tested. The comparison of elderly normal with young normal showed that A-wave acceleration did not change significantly. Comparing elderly normal to elderly with diseases showed that all tested indexes except A wave acceleration failed to separate normal elderly from diseased elderly. A-wave acceleration appears to be a useful index that can help in diagnosis of left ventricular diastolic dysfunction in elderly patients independent of age effects. PMID- 18388034 TI - Hierarchical analysis of cardiovascular risk factors in relation to the development of acute coronary syndromes, in different parts of Greece: the CARDIO2000 study. AB - During 2000 to 2002, 700 men (59 +/- 10 years) and 148 women (65 +/- 9 years) patients with first event of an ACS were randomly selected from cardiology clinics of Greek regions. Afterwards, 1078 population-based, age-matched and sex matched controls were randomly selected from the same hospitals. The frequency ratio between men and women in the case series of patients was about 4:1, in both south and north Greek areas. Hierarchical classification analysis showed that for north Greek areas family history of coronary heart disease, hypercholesterolemia, hypertension, diabetes (explained variability 35%), and less significantly, dietary habits, smoking, body mass index, and physical activity status (explained variability 4%) were associated with the development of ACS, whereas for south Greek areas hypercholesterolemia, family history of coronary heart disease, diabetes, smoking, hypertension, dietary habits, physical activity (explained variability 34%), and less significantly body mass index (explained variability <1%), were associated with the development of the disease. PMID- 18388035 TI - Potential pitfalls of renal angiography: a case of atypical fibromuscular dysplasia. AB - Although angiography remains the gold standard for the diagnosis of renal artery stenosis, renovascular disease can at times present with unusual patterns that may be difficult to detect. The authors present a case in which an initial renal angiogram failed to identify the presence of severe disease involving both a main and accessory renal artery. Repeat angiography coupled with the use of adjunctive catheter-based techniques including translesional pressure gradient determination and intravascular ultrasound with virtual histology imaging revealed the presence of atypical fibromuscular dysplasia that was treated with good clinical results. The case highlights the importance of performing careful and complete renal angiography, including imaging of smaller accessory renal arteries, and describes several readily available catheter-based techniques that can be useful in elucidating the physiological significance and etiology of renal artery stenosis. PMID- 18388037 TI - Association between protein-bound sialic acid and high-sensitivity C-reactive protein in essential hypertension: a possible indication of underlying cardiovascular risk. AB - The aim of this study was to examine the possible alteration in the levels of C reactive protein, protein-bound sialic acid, and other lipid risk factors in newly diagnosed essential hypertensive subjects. In all, 56 hypertensive and 33 normotensive male subjects were enrolled in the study. Lipid profile, C-reactive protein, apolipoprotein-B, and protein-bound sialic acid were estimated in both the groups. Total cholesterol, triglyceride, low-density lipoprotein-cholesterol, C-reactive protein, apolipoprotein-B, and protein-bound sialic acid were significantly increased in patients with essential hypertension. Correlation analysis revealed a significant association between the protein-bound sialic acid with mean arterial pressure, C-reactive protein, and low-density lipoprotein cholesterol. The findings of the present study suggest that in essential hypertension there is an association between protein-bound sialic acid and C reactive protein, which reflects the clustering of cardiovascular risk factors in these patients. PMID- 18388036 TI - Ceruloplasmin is a better predictor of the long-term prognosis compared with fibrinogen, CRP, and IL-6 in patients with severe unstable angina. AB - BACKGROUND: We investigated the time course and prognostic value of fibrinogen (Fib), C-reactive protein (CRP), interleukin-6 (IL-6), and ceruloplasmin (CP) in patients with severe unstable angina. METHODS: All 4 substances were measured on admission and after 6, 12, 24, 48, and 72 hours, and after 7 days and 6 months in 40 patients with Braunwald's classification class IIIB unstable angina. RESULTS: All recorded substances increased significantly; 15 patients had cardiovascular events during hospitalization and 11 patients during follow-up. The time course and the mean values of Fib, CRP, and IL-6 were similar in patients with and without complications both during hospitalization and follow-up. However, CP levels from 6 hours until 6 months were significantly higher in patients with complications during follow-up (P < .05). CONCLUSIONS: Fib, CRP, IL-6, and CP levels alter in patients with severe unstable angina. However, only CP levels were related to 12-month follow-up prognosis. PMID- 18388038 TI - Effects of policosanol (10 mg/d) versus aspirin (100 mg/d) in patients with intermittent claudication: a 10-week, randomized, comparative study. AB - Antiplatelet therapy, including aspirin, is recommended to lower the vascular risk in patients with intermittent claudication. Policosanol has increased walking distances in these patients, probably because of its antiplatelet effects, but the effect of shorter treatment has not been studied. This double blind study compared the effects of policosanol 10 mg/d and aspirin 100 mg/d for 10 weeks on walking distances in claudicants. Thirty-nine patients were randomized to policosanol or aspirin. Walking distances on a treadmill were assessed before and after treatment. Policosanol significantly increased the initial and absolute claudication distances, while aspirin changed neither variable. Policosanol, not aspirin, significantly lowered serum low-density lipoprotein-cholesterol and total cholesterol while raising high-density lipoprotein-cholesterol. In conclusion, treatments of policosanol, not aspirin, for 10 weeks significantly increased walking distances, but modestly, in contrast with previous results. Therefore, the duration of treatments at the doses tested was too short for meaningful effects on the treadmill test. PMID- 18388040 TI - Leiomyosarcoma of the inferior vena cava: a case report. AB - Leiomyosarcoma of the inferior vena cava is an extremely rare tumor that is characterized by a poor prognosis and nonspecific symptoms, a fact that may delay the diagnosis for several years. The only therapeutic modality proven to prolong the survival of patients is total surgical resection of the tumor. In this study, the authors report the case of a 50-year-old patient with a diagnosis of leiomyosarcoma of the inferior vena cava, affecting the middle and distal thirds, who was submitted to surgical treatment. PMID- 18388039 TI - Thienopyridine therapy influences late outcome after coronary stent implantation. AB - Clinical significance of resistance to aspirin and thienopyridine therapy is poorly defined. The authors aimed to evaluate whether more effective antiplatelet therapy is associated with better outcome in patients on dual-antiplatelet treatment. Using optical aggregometer, maximal platelet aggregation values were measured with induction of adenosine diphosphate, collagen, and adrenaline 30 +/- 5 days after coronary stent implantation in 134 patients. Markers of platelet activation were also analyzed with fluorescent immunoassay in 57 patients. After 10 months of follow-up, 33 patients reached the composite endpoint of cardiovascular death, myocardial infarction, and revascularisation. Adenosine diphosphate-induced maximal aggregation values were in significant relationship with the development of major adverse cardiac events (P < .01). Level of soluble P-selectin proved to be an independent risk factor of adverse outcome (P < .05). As efficacy of thienopyridine therapy showed significant relation with clinical outcome, the authors conclude that interindividual variability in response to adenosine diphosphate-receptor antagonists may be of substantial clinical importance. PMID- 18388041 TI - Assessment of early changes in the segmental functions of the left and the right ventricles after biventricular pacing in heart failure: a study with tissue Doppler imaging. AB - Tissue Doppler imaging allows assessment of systolic and diastolic regional ventricular function. The aim of this study was to assess early changes in regional systolic and diastolic functions and differences in transition time to contraction between the ventricles after cardiac resynchronization therapy. Fourteen patients were included, who underwent echocardiography before and 1 month after resynchronization. The difference between transition time to contraction of left and right ventricles decreased to 24.4 +/- 10.7 milliseconds from 65.3 +/- 18.2 milliseconds after resynchronization therapy (P = .001). There was a significant relation between the decrease in difference between transition time and increase in ejection fraction (r = 0.80, P = .002). Early or late diastolic myocardial motion increased in 7 segments of left and 2 segments of right ventricles. Systolic myocardial motion increased in 7 segments of left and in all segments of right ventricles. Resynchronization therapy improved systolic and diastolic functions in both ventricles. The difference between transition time to contraction of ventricles might be helpful in estimating optimal resynchronization. PMID- 18388042 TI - Life-threatening pacemaker dysfunction associated with therapeutic radiation: a case report. AB - Reports about pacemaker (PM) dysfunction during irradiation (IR) are very rare, which is because of the extensive protective mechanisms that exist in these devices against electromagnetic interference (EMI). We report a case in which one of the most clinically relevant type of PM malfunctions, a runaway PM, occurred during radiation in a 76-year-old woman who was treated for inoperable esophageal cancer with a course of photon IR. The estimated IR dose of 0.11 Gy was the lowest in vivo dose ever reported. So a direct radiation effect as cause for this malfunction appears to be improbable. It could be concluded that the PM dysfunction was most likely induced by EMI during radiotherapy. The real reason of the device's software failure remains unclear. PMID- 18388043 TI - The roles of oxidized low-density lipoprotein and interleukin-6 levels in acute atherothrombotic and lacunar ischemic stroke. AB - The role of circulating, oxidized low-density lipoprotein and interleukin-6 levels in acute ischemic stroke considering the primary-vessel disease was investigated. The study consisted of 28 patients with acute ischemic stroke and 23 control subjects. Patients were subdivided into large-vessel (n = 12) and small-vessel (n =16) disease stroke groups according to the Trial of Org 10172 in Acute Stroke Treatment (TOAST) criteria. The means of oxidized low-density lipoprotein and interleukin-6 levels of patients with acute ischemic stroke were higher than controls (P < .01, P < .05). Mean oxidized low-density lipoprotein level was higher in the large-vessel disease group than in the small-vessel disease group (P < .01). The mean of inteleukin-6 levels was higher in the small vessel disease group (P < .01). The results of the present study showed that oxidative stress promotes large-vessel disease rather than small-vessel disease stroke, and inflammation may play important an role in the development of small vessel disease stroke. PMID- 18388044 TI - Chronic effort-induced angina as presentation of a totally occluded left main coronary artery: a case report and review. AB - In this report, the case of a 40-year-old patient with chronic total occlusion of the left main coronary artery is discussed. Total occlusion of the left main coronary artery is a rare finding at coronary angiography. When present, patients most often have extensive collateral circulation from the right coronary artery. The mainstay of treatment is surgical with coronary artery bypass grafting. PMID- 18388045 TI - Late angiographic stent thrombosis: the LAST straw for drug-eluting stents? AB - The long-term patency advantage of drug-eluting stents represents a quantum leap in the percutaneous treatment of ischemic heart disease. Although initial landmark trials demonstrated equivalent safety to bare-metal stents, subsequent follow-up analyses have suggested a slight late thrombotic risk. This widely publicized issue poses major public health implications for the medical and lay communities. However, available data indicate that this late risk is counterbalanced by the dramatic drug-eluting stent reduction in target lesion revascularizations, resulting in equivalent overall major adverse cardiovascular event rates than that of the bare-metal stents The recent Food and Drug Administration's guidelines regarding these devices are delineated in detail. Specifically, compliance with dual antiplatelet therapy (minimum 1 year) is of paramount consideration in patient selection for drug-eluting stents. Moreover, when deployed for "on-label" indications, they provide significant restenosis advantage, with a slight late thrombotic risk but without overall increase in death or infarction. PMID- 18388046 TI - Myocardial perfusion in patients with a totally occluded left anterior descending coronary artery reinjected by a normal right coronary artery: the role of collateral circulation. AB - In this article, myocardial perfusion in patients with a totally occluded left anterior descending artery reinjected by a normal right coronary artery is assessed using stress single photon emission computed tomography (SPECT). In all, 20 patients, with a totally occluded left anterior descending artery reinjected by normal right coronary artery, underwent myocardial single photon emission computed tomography imaging within 60 days of angiography. All patients had abnormal perfusion single photon emission computed tomography results and 70% had reversible defects. Perfusion defects at rest were present in 75% of patients, with perinecrotic residual ischemia in 45% of patients whereas for 30% of patients, no viable myocardium was detected in the collateral-dependent segments. In all, 25% of patients had no resting perfusion defects but all are presented with stress-induced ischemia. Collaterals are not protective against stress induced ischemia, but they can preserve myocardial viability. This conclusion is highly supported by the presence of residual ischemia in the collateral-dependent segments. PMID- 18388047 TI - Acute posttraumatic ischemia of the limbs: algodystrophy or related syndrome? A prospective study on a series of 25 patients. AB - The aim of this study was to present clinical and paraclinical features of a rarely studied vascular syndrome that usually occurs after a minor traumatic injury. The prospective study of 25 patients identified a group of young subjects, mainly women, generally with history of vascular hyperactivity to cold, showing disseminated pain in a limb always combined with vasospasm, consecutive to a minor traumatic injury in general, and simulating acute ischemia of a limb. The patients' arterial and microcirculatory flows were restored after warming, which proved that they were suffering from intense, reversible but pathological vasospasm, because of its duration, diffusion, intensity, and disproportion as to the triggering event. The symptoms may recur, but they respond to physiotherapy. Patients' follow-up showed that microcirculatory function remains abnormal sometime after the initial episode with, in particular, an exacerbation of the venoarteriolar reflex. PMID- 18388048 TI - Acute pancreatitis mimicking acute inferior myocardial infarction. AB - A 56-year-old man presented with acute pancreatitis and electrocardiographic abnormalities, suggesting acute inferior myocardial infarction. An emergent coronary angiogram showed normal coronary arteries. The clinical significance and therapeutic implications of this rare finding are discussed. PMID- 18388049 TI - Traumatic dissection of the left circumflex artery after a fall from height. AB - The authors present the case of a healthy 40-year-old man who developed a myocardial infarction because of traumatic dissection of the left circumflex coronary artery following a fall from height. To our knowledge, this is the first time that a traumatic lesion of the left circumflex artery is being reported after a fall. PMID- 18388050 TI - Primary myxofibrosarcoma of the left atrium: case report and review of the literature. AB - A 63-year-old woman with progressive dyspnea underwent transthoracic echocardiography and was found to have a large multilobed mass in the left atrium that was attached to lateral wall. On inspection during surgery, the tumor was found to infiltrate the posterior mitral annulus and leaflet. The patient underwent surgical resection of the tumor and mitral valve replacement. Histologic and cytochemical evaluation confirmed that the tumor was a myxofibrosarcoma. Despite chemotherapy, the tumor recurred and the patient died 3 months after surgery. PMID- 18388051 TI - Ephedrine-induced acute myocardial infarction in a young athlete: a case of thrombus management. AB - A case of acute myocardial infarction in a young athlete provoked by ephedrine abuse has been described in this study. An intracoronary thrombus found in the left anterior descending coronary artery at urgent angiography was successfully removed using the Pronto (Vascular Solutions, Minneapolis, Minnesota) aspiration catheter. The intravascular ultrasound examination performed thereafter showed a nonobstructive atherosclerotic plaque in the culprit artery; there was no evidence whatsoever of possible plaque disruption. The result of percutaneous coronary intervention was satisfactory, and no stent implantation was needed. The patient experienced no adverse events until his outpatient visit 3 months later. PMID- 18388052 TI - Tako-tsubo-like syndrome with atypical clinical presentation: case report and literature review. AB - A 78-year-old woman presented herself at emergency for the appearance of severe dyspnea. An electrocardiogram showed signs of inferior and anterior necrosis, and laboratory tests showed a small increase of myocardial cytonecrosis enzymes. An echocardiogram detected a reduction of global systolic function (ejection fraction [EF] approximately 40%) as well as akinesia of the apex, interventricular septum middle segments, and anterior and anterolateral walls, with basal segments showing compensatory hyperkinesia. The coronarography showed a coronary tree substantially free from significant lesions. The patient was transferred to the cardiology unit of our hospital. Based on the contained increase of the cardiac enzymes, the absence of coronary lesions, and the presence of typical echocardiography alterations, we suspected a Tako-tsubo-like syndrome. On deeper anamnestic investigation, an event of strong emotional stress emerged preceding the hospital admission that confirmed the pathology, even though it is atypical to see clinical presentation a long time after a stressful event. PMID- 18388053 TI - The effects of different beta-blockers on left-ventricular volume and function after primary coronary stenting in acute myocardial infarction. AB - The beneficial effect of beta-blockers is reported in myocardial infarction (MI). This study compared the effects of propranolol and carvedilol on left-ventricular (LV) volume and function after acute MI. Serial echocardiographic studies were performed on acute MI patients who were treated with primary coronary stenting and medical treatment, including propranolol or carvedilol. Determinants of the changes in LV volume and function were identified by regression analyses. At 6 months, there was no significant difference in change in the LV end-diastolic volume between patients receiving 2 beta-blockers. The increase of LV ejection fraction was greater in patients receiving carvedilol. On multivariate analyses, it was found that the kind of beta-blocker had no significant influence on the changes in LV volume or ejection fraction, whereas gender and baseline LV ejection fraction were predictive of change in LV ejection fraction. Compared with propranolol, carvedilol did not have a significant benefit on LV remodeling or function after primary coronary stenting in acute MI. PMID- 18388054 TI - The optimal time of elective percutaneous coronary intervention for stable patients after ST elevation myocardial infarction. AB - OBJECTIVE: To find the optimal time (early: < or =3 days; late: >3 days) for revascularization in ST elevation myocardial infarction (STEMI) patients in the subacute phase. METHODS: Ninety-nine STEMI patients who were admitted to Gazi University Faculty of Medicine between 2000 and 2004 were enrolled into this study. Patients were divided into 2 groups according to time from the beginning of symptoms to the percutaneous coronary intervention. Coronary angiograms before and after the revascularization were evaluated using the quantitative coronary angiogram technique. RESULTS: 45 early (group I) and 54 late (group II) revascularized patients were evaluated. There were no significant differences between the 2 groups regarding demographic properties, thrombus score, success of the procedure, quantitative angiographic parameters, and clinical results of the procedure. CONCLUSIONS: Waiting for the development of stable phase in STEMI to apply PCI has no obvious benefit for angiographic and clinical results. PMID- 18388055 TI - Dynamic nutcracker-like compression of saphenous vein graft by adjacent stents. AB - Systolic compression of coronary flow is a rare angiographic finding and usually results from intramyocardial bridging. In this article, the first reported case of iatrogenic dynamic saphenous vein graft compression by adjacent stents is presented. The cyclic movements of the stents resemble the action of a nutcracker with the involved segment at the fulcrum. Previous angiography 4 months prior showed no evidence of such flow impairment, thereby excluding intramyocardial bridging as a potential etiology. Progressive scarring and fibrosis may result in adhesion of a noncoronary conduit to a stationary mediastinal structure. Thus, conceptually, such vessels would be more prone to systolic compression and torsion, as they do not move with the cardiac cycle. Although the stent deployment may be beneficial in some scenarios, complications of vessel rupture and stent collapse can occur. The pathophysiology, natural history, and treatment of dynamic vessel compression are briefly reviewed. PMID- 18388056 TI - Photoplethysmographic assessment of hemodynamic variations using pulsatile tissue blood volume. AB - Sympathetic responses to provocative tests have shown to provide an early prognosis of abnormalities in the human autonomic nervous system. Photoplethysmographic signal characteristics have been studied to identify vascular diseases. However, knowledge about the pulse-added arterial volume of the photoplethysmographic waveforms during these clinical investigations is limited. In all, 16 normotensive adults (9 men) were recruited to perform 3 provocative test (2 postural changes and 1 resistive breathing) activities with photoplethysmographic signals being monitored on the upper and lower peripheries using customized devices. In all, 4 parameters derived from the photoplethysmographic waveforms that related to the pulsatile tissue blood volume changes were then assessed. The results obtained showed that amplitude-related parameters during these activities showed significant changes (>9.63%; P < .05). However, comparison of pulse-added arterial volume showed insignificant changes (<5.56%; P > .05) for all test settings. Hence, the findings herein suggest that there is clinical potential in using this aspect of the photoplethysmographic waveform. PMID- 18388057 TI - Elevated osteopontin levels in patients with peripheral arterial disease. AB - This study was carried out to compare concentrations of osteopontin (OPN) and osteoprotegerin (OPG) in peripheral arterial disease (PAD). The study population consisted of 200 consecutive subjects in whom both OPN/OPG and ankle-brachial index were measured. It was found that OPN levels, but not OPG levels, were significantly more increased in patients with PAD than those without PAD. Serum OPN levels were significantly lower in subjects with angiotensin converting enzyme inhibitors or angiotensin II receptor blockers than those without these agents. In this study, it has been demonstrated for the first time that serum OPN levels are related to PAD. Inhibition of renin- angiotensin system could decrease OPN levels and prevent the progression of PAD. PMID- 18388058 TI - Ostial lesions of left main and right coronary arteries: demographic and angiographic features. AB - In 258 patients with left main tract disease, the atherosclerotic risk factors were compared between patients with ostial and nonostial lesions of the left main coronary artery. Also, it was done for patients with ostial right coronary artery. Women were more likely to have ostial left main coronary artery and/or ostial right coronary artery. A multivariate logistic regression analysis revealed that the female sex (odds ratio: 2.336) and hypertriglyceridemia (odds ratio: 1.004) were independent risk factors of ostial left main coronary artery lesion. For ostial right coronary artery lesion, the female sex and family history of coronary artery disease were independent predictors. Ostial left main coronary artery and right coronary artery lesions were strongly correlated. The demographic and clinical profiles of ostial stenosis suggest that this group may represent a distinct entity, different from the more common atherosclerotic left main trunk stenosis (LMTD). The female sex and serum triglyceride level can be considered as independent predictors of ostial left main tract disease. PMID- 18388059 TI - Management of primary pulmonary artery sarcoma: experience of a single center. AB - Primary sarcoma of the pulmonary artery (PSPA) is extremely rare. Many cases are misdiagnosed as pulmonary arterial hypertension (PAH) because of chronic thromboembolic disease (CTD). Four cases of PSPA with the initial misdiagnosis are reported. The presence of a unique mass in the main pulmonary artery or proximal branches, rapidly progressive dyspnea, and constitutional symptoms should raise the suspicion of PSPA. The pathological diagnosis is usually confirmed during surgery, which is done along with adjuvant chemotherapy, the treatment of choice. PMID- 18388060 TI - Flow-mediated arterial dilation in primary antiphospholipid syndrome. AB - OBJECTIVE: To investigate precocious alterations in the artery wall of patients with primary antiphospholipid syndrome (APS). METHODS: We evaluated flow-mediated dilation (FMD) of the brachial artery and intima-media thickness (IMT) of carotid arteries in 16 patients and 16 healthy controls matched for age, gender, and other cardiovascular risk factors. RESULTS: FMD of the brachial artery was significantly lower in patients than in controls (6.3 +/- 5.2% vs 18.2 +/- 2.7; P < .005). IMT was similar in the 2 groups. FMD was significantly reduced in patients with anticardiolipin antibodies IgM. CONCLUSIONS: APS correlates closely with precocious atherosclerosis, and the correlation with a type of anticardiolipin antibody may be predictive of more accelerated atherosclerosis. PMID- 18388061 TI - Secondary prevention of arterial disease in very elderly people: results from a prospective registry (FRENA). AB - There is little information on the effectiveness of secondary prevention interventions in very elderly patients. In this article, the incidence of major cardiovascular events during a 12-month follow-up period in a series of consecutive patients with coronary, cerebrovascular, or peripheral artery disease is analyzed. As of October 2006, 1264 patients had been enrolled. Of these, 324 (26%) were >or=75 years of age. Their incidence rate of 22 events per 100 patient years (95% CI, 17-28) was over 2-fold the 7.9 (95% CI, 6.2-10) found in those <75 years of age. Among them, only chronic heart failure and diabetes were independently associated with an increased risk for major events, whereas the use of angiotensin II antagonists was associated with a lower risk. Patients >or=75 years of age had an over 2-fold higher incidence of major cardiovascular events. The use of angiotensin II antagonists was associated with a lower risk. PMID- 18388062 TI - Acute myocardial infarction in a young man using anabolic steroids. AB - Anabolic-androgenic steroids are used worldwide to help athletes gain muscle mass and strength. Their use and abuse is associated with numerous side effects, including acute myocardial infarction (MI). We report a case of MI in a young 31 year-old bodybuilder. Because of the serious cardiovascular complications of anabolic steroids, physicians should be aware of their abuse and consequences. PMID- 18388063 TI - Lipid-lowering therapy is related to inflammatory markers and 3-year mortality in patients with critical limb ischemia. AB - To evaluate relationships between lipid-lowering therapy, inflammation, and 3 year mortality in critical limb ischemia (CLI), 259 consecutive CLI patients underwent evaluation of medication, tumor necrosis factor-alpha, interleukin-6 (IL-6), neopterin, high-sensitivity C-reactive protein (hs-CRP), 8-epi-PGF(2 alpha), and endothelin-1. Mortality was assessed after 3 years. Sixty-one patients (24%) were on lipid-lowering therapy and 59 patients (97%) on statins. Patients on lipid-lowering therapy were younger and showed lower low-density lipoprotein cholesterol, hs-CRP, and IL-6 levels than patients without therapy. Three-year survival was higher among patients on lipid-lowering therapy. In logistic regression, the effect of lipid-lowering therapy on 3-year survival was significant with inflammatory markers entered into the model one by one but disappeared when all inflammatory markers were entered into the model together. In conclusion, hs-CRP and IL-6 levels were lower and 3-year survival was higher in CLI patients on lipid-lowering therapy. PMID- 18388064 TI - Coronary collateral circulation: any effect on P-wave dispersion? AB - Coronary collateral circulation determines the severity of ischemic myocardial damage. Increased P-wave dispersion is an independent predictor for atrial fibrillation. Consistent evidence is little about the relation between coronary collateral circulation and arrhythmia risk. In this article, the effect of coronary collateral circulation on P-wave dispersion was evaluated. Collateral grade and P-wave dispersion were ascertained in 100 patients with >or=85% diameter stenoses in left anterior descending or right coronary arteries. Left ventricular function score was also determined in all patients. Coronary collateral circulation was absent in 32 patients, whereas 68 patients had coronary collateral circulation. Patients with collateral grade >or=1 had greater left ventricular function score than did patients with collateral grade 0 (P = .048). However, there was no significant difference between P-wave dispersion of patients with and without coronary collateral circulation (P = .45). The presence of coronary collateral circulation failed to exert a beneficial decreasing effect on P-wave dispersion. PMID- 18388065 TI - Assessment of left ventricular functions in patients with isolated coronary artery ectasia by conventional and tissue Doppler imaging. AB - The authors sought to determine left ventricular functions by conventional and tissue Doppler imaging in patients with isolated coronary artery ectasia and controls. Peak early (E) and late (A) mitral inflow velocity, E/A ratio, E deceleration time, and isovolumetric relaxation time were obtained. Peak systolic velocity (Sm), diastolic early (Em), and late (Am) velocities were measured by tissue Doppler imaging. Interventricular septum velocities, including peak systolic (Ss), diastolic early (Es), and late (As) velocities, were recorded. Peak early (E) velocity, E/A ratio, and E deceleration time were different in both groups. Isovolumetric relaxation time was prolonged in patients with coronary artery ectasia than controls. Em and Em/Am ratio were lower in patients with coronary artery ectasia than controls. Diastolic early and Es/As velocities were lower in patients with coronary artery ectasia compared with controls. The authors showed that mitral inflow-lateral annulus and interventricular septum velocities were lower in patients with coronary artery ectasia than controls indicating left ventricular diastolic dysfunction. PMID- 18388066 TI - Simultaneous presentations of deep vein thrombosis and cerebral sinus thrombosis in a case of primary antiphospholipid syndrome. AB - Antiphospholipid syndrome is characterized with arterial and venous thrombosis. In this article, a 38-year-old man presented with headache and swelling of his left limb, which had lasted for 2 months. Duplex ultrasonography showed thrombosis of the left femoral vein. Cerebral magnetic resonance imaging also showed cerebral sinus thrombosis. Serological examination showed that antiphospholipid syndrome was the underlying disease condition in this patient. Despite adequate anticoagulant therapy, deep vein thrombosis and cerebral sinus thrombosis exacerbated, resulting in inferior vena cava occlusion, papilloedema, and abducent nerve paralysis. Optic canal decompression and cistern-peritoneal shunt operation were performed, following which his neurological symptoms were relieved. The occurrence of cerebral sinus thrombosis as initial presentation of antiphospholipid syndrome is extremely rare and remains a diagnostic challenge. Although the clinical presentation is highly variable, the diagnosis should be considered in patients with antiphospholipid syndrome presenting with recent unusual headaches. Improved diagnosis and treatment strategy may ultimately improve the clinical outcome of these patients. PMID- 18388067 TI - Relation of serum uric acid levels with the presence and severity of angiographic coronary artery disease. AB - The aim of this study is to investigate the association between uric acid level and severity of coronary artery disease. Consecutive 495 patients with coronary artery disease and 356 individuals with normal coronary angiograms were included in the study. Severity of coronary artery disease was evaluated using the Gensini score index. For both groups, conventional risk factors, the levels of uric acid, and other biochemical markers were assessed. The mean uric acid levels of the patient group were significantly higher than those of the control group (P = .002). Mean uric acid levels were higher in men than in women (P < .001). In the coronary artery disease group, there was no significant correlation between uric acid levels and Gensini score in both sexes. Thus, it can be concluded that uric acid level is associated with the presence but not with the severity of coronary artery disease. PMID- 18388068 TI - Risk stratification of apolipoprotein B, apolipoprotein A1, and apolipoprotein B/AI ratio on the prevalence of the metabolic syndrome: the ATTICA study. AB - We investigated the association of apolipoproteins AI and B in relation to the prevalence of metabolic syndrome in a random sample of cardiovascular disease- free adults from the ATTICA study (1,514 men, aged 18-87 y; 1,528 women, aged 18 89 y). Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III criteria. The prevalence of metabolic syndrome was 25% in men and 15% in women (P < .001). Using the area under the Receiver Operation Characteristic curve, apolipoprotein B/AI was the best diagnostic marker of metabolic syndrome, the optimal discriminating cut-off value of this ratio was 0.72 (sensitivity 74%, specificity 67%), and individuals with apolipoprotein B/AI ratio greater than 0.74 had 3.29 times higher odds of having metabolic syndrome (95% confidence interval: 2.56-4.21) after adjusting for potential confounders. PMID- 18388069 TI - Relationships between vascular indexes and atherosclerotic risk factors in patients with type 2 diabetes mellitus. AB - The relationships of cardio-ankle vascular index, (a new indicator of arterial stiffness), common carotid artery intima-media thickness, and ankle-brachial pressure index with atherosclerotic risk factors were compared in patients with type 2 diabetes. There were significant correlations between each pair of the above vascular indexes. Both cardio-ankle vascular index and intima-media thickness showed significant correlations with age, systolic blood pressure, pulse pressure, and serum total cholesterol. Ankle-brachial pressure index showed a significant negative correlation with pulse pressure but not with systolic blood pressure and total cholesterol. Logistic regression analysis showed that cardio-ankle vascular index and intima-media thickness were associated with systolic blood pressure and pulse pressure independently of age. These results suggest that there are significant associations among the 3 vascular indexes and that systolic blood pressure and pulse pressure are major, age-independent determinants of cardio-ankle vascular index and common carotid artery intima media thickness in patients with diabetes. PMID- 18388070 TI - Splenic vein thrombosis following abdominal compression and vibration: a case report. AB - The authors describe a 46-year-old man with isolated splenic vein thrombosis (SVT) that developed after concrete drilling, which caused significant compression and vibration of the abdominal wall. Methyltetrahydrofolate reductase (MTHFR) deficiency-related hyperhomocysteinemia, which was subsequently diagnosed, predisposed him to this event. To the best of the knowledge of authors, this is the first report of isolated splenic vein thrombosis caused by compression and vibration of the abdomen. Relevant aspects of isolated post traumatic splenic vein thrombosis in this context are discussed. PMID- 18388071 TI - Gigantic bronchial artery aneurysm treated with transcatheter arterial embolization: a case report. AB - Bronchial artery aneurysm (BAA) is a rare condition. Rupture of BAA can cause critical hemorrhage, and intervention for BAA is thus recommended. A 69-year-old woman presented with BAA 70 mm in diameter in the right hilum of the lung. Transcatheter arterial embolization for afferent arteries of the BAA was performed and the BAA has subsequently been shrinking as observed by radiography. We present this case and a brief review of management of BAA based on the literature. PMID- 18388072 TI - Endothelial dysfunction in chronic obstructive pulmonary disease. AB - BACKGROUND: Cardiovascular diseases are prevalent in people with chronic obstructive pulmonary disease (COPD). We hypothesized that endothelial dysfunction could be a marker of the proatherogen status in COPD. METHODS AND RESULTS: We measured endothelial dysfunction by flow-mediated dilation (FMD) and after sublingual administration of nitroglycerin (nitrate-mediated dilation: NMD) in 44 COPD patients and 48 controls. Compared with controls COPD patients had worse mean FMD (5.4% vs 8.2%, P < .001) and NMD (12.0% vs 13.9%, P = .007). FMD was inversely related to FEV1/VC ratio (r = -0.327, P = .030). The negative association between COPD and FMD was confirmed after correction for potential confounders in a multiple linear regression model (beta = -0.019, P = .002). In the same model NMD (beta = 0.396, P < .001) was positively associated with FMD. CONCLUSIONS: Endothelial-dependent and, to a lesser extent, endothelial independent dilations are significantly impaired in COPD, and the impairment is proportional to the severity of bronchial obstruction. PMID- 18388073 TI - Association between lower-extremity function and arterial compliance in older adults. AB - The purpose was to identify the association between lower-extremity function and arterial compliance in older men and women. Participants included 46 healthy men (n = 18) and women (n = 28) 60 years of age or older. Lower-extremity functional performance was assessed by the summary performance score (SPS) that includes tests of 5 timed repeated chair rises, standing balance, and 4-meter walking velocity. Arterial compliance and arterial pressure were analyzed through pulsewave analysis. Small arterial compliance (3.74 +/- 2.14; mean +/- SD) was related (r = 0.34, P = .028) to SPS (11.09 +/- 1.19) after adjusting for body surface area, hyperlipidemia, and hypertension. Systolic blood pressure (138 +/- 14) also was related to SPS (r = -0.314, P = .040). These results suggest diminished lower-extremity function is associated with decreased small arterial compliance and elevated arterial pressure in older men and women. PMID- 18388075 TI - Progression of nonculprit plaque stenosis following successful percutaneous intervention. AB - Despite percutaneous intervention after an acute coronary syndrome, patients remain at high risk for recurrent events in the first year. Prior studies have shown that a plaque rupture can occur not only at a single culprit lesion site but also in other atherosclerotic plaques throughout the coronary vasculature in patients with stable angina, silent myocardial ischemia, and during acute coronary syndromes. A 71-year-old man who presented with exertional angina and who had a successful stent in a culprit right coronary artery is described in this article. After 4 weeks, he represented with accelerated angina. A prior lesion in the obtuse marginal artery, remote from the site of the index lesion, had progressed from a 30% to 90% stenosis. This case report demonstrates the systemic nature of acute coronary syndromes, highlights the inherent instability of coronary artery disease, and supports the notion of aggressive secondary prevention in these patients. PMID- 18388074 TI - Catastrophic antiphospholipid syndrome presenting with multiorgan failure and gangrenous lesions of the skin. AB - A 38-year-old woman with a 20-year history of systemic lupus erythematosus and positive anticardiolipin antibodies developed anuria and hypotension. 20 days before, she had discontinued 25 mg prednisone, but not warfarin, on medical advice. 3 days before admission, she developed extensive necrosis and blisters involving both arms and legs and a multiorgan failure. She improved after immunosuppressive and antibiotic therapy, 3 sessions of dialysis and 5 sessions of plasmapheresis. It was decided that she could be discharged after 45 days. Her skin lesions were complicated by several abscesses requiring surgical drainage and finally healed almost completely within 9 months. Catastrophic antiphospholipid syndrome is a distinctly rare dramatic condition characterized by widespread thrombosis of small vessels, which in the present case was likely triggered by the abrupt withdrawal of steroid therapy. It should be borne in mind in cases of multiorgan failure, which does not recognize a well-defined etiology. PMID- 18388076 TI - Spontaneous coronary artery dissection mimicking acute aortic dissection. AB - Spontaneous coronary artery dissection is a rare cause of acute coronary syndrome or sudden cardiac death, occurring predominantly in the female sex without cardiovascular risk factors. Although the etiology and pathogenesis remain uncertain, hypertension has not appeared to be one of the most important factors. This case report describes a patient with spontaneous coronary artery dissection in proximal right coronary artery who presented the signs and symptoms of acute aortic dissection. The authors postulate that hypertension may have been an important factor for the pathogenesis of the spontaneous coronary artery dissection. PMID- 18388077 TI - Late presentation of a lead perforation as a complication of permanent pacemaker insertion. AB - Lead perforation is a less-recognized delayed complication of device implantation. Delay in recognition may prove fatal. Predictors of postimplantation pericardial effusion, which serves as a marker of perforation, include concomitant use of transvenous pacemaker, steroid use within 7 days, and older age. The authors report a case of an 86-year-old patient who presented with a lead perforation 16 months following the original pacemaker insertion. Following surgical repair with sternotomy, a new ventricular lead was placed via a transvenous approach at the right ventricular septum. A higher clinical suspicion should be maintained in the elderly in whom perforation occurs more frequently, and consideration should be given to implanting the lead in sites other than the right ventricular apex to minimize the risk of this late complication. PMID- 18388078 TI - "Perilous pearl"--papillary fibroelastoma of aortic valve: a case report and literature review. AB - Papillary fibroelastomas of the heart valves are benign, slow-growing, rare tumors of the heart. These lesions are primarily responsible for embolic events that can clinically manifest with neurological and cardiovascular symptoms. Early diagnosis is very important, as surgical excision of these tumors can prevent cerebrovascular and cardiovascular complications. The case of a 60-year-old man who presented with a neurological deficit caused by a papillary fibroelastoma of the noncoronary cusp of the aortic valve is described. Diagnosis was made by transesophageal echocardiogram, and the tumor was resected surgically. PMID- 18388079 TI - Percutaneous thrombin injection for treatment of iatrogenic femoral artery pseudoaneurysms: a case for caution. AB - The case of a patient who developed a femoral artery pseudoaneurysm following cardiac catheterization is described. After 2 failed attempts of ultrasound guided compression repair, the patient underwent percutaneous thrombin injection with, beside of complete closure of the pseudoaneurysm, a severe limb ischemia due to distal thrombin migration with consecutive clot formation finally resulting in thigh amputation of the affected leg. Indications, advantages, and disadvantages of various options for the treatment of iatrogenic femoral artery pseudoaneurysms (vascular surgery, ultrasound-guided compression, percutaneous thrombin injection, and other nonsurgical treatment modalities), as well as risk factors for distal migration of liquid thrombin after percutaneous injection, are discussed in this report. PMID- 18388080 TI - Fate of incidental, asymptomatic lesions discovered during percutaneous coronary intervention. AB - The authors aimed to determine the incidence and angiographic features associated with plaque progression requiring nontarget lesion percutaneous coronary intervention after culprit lesion percutaneous coronary intervention. Of 945 consecutive percutaneous coronary interventions reviewed, 100 patients who required nontarget lesion percutaneous coronary intervention in the following year of the index percutaneous coronary intervention were found and compared with 100 consecutive patients who did not require nontarget lesion percutaneous coronary intervention. Patients with restenosis were excluded. Incidence of clinical plaque progression leading to additional nontarget lesion percutaneous coronary intervention in the year after an index percutaneous coronary intervention was found to be 10.5%. In multivariable logistic regression analyses, the predictors of plaque progression were multivessel disease, unstable angina pectoris, diabetes mellitus, prior percutaneous coronary intervention, and lack of statin use. Initially, lesions that lead to repercutaneous coronary intervention were mostly nonsignificant. Clinical presentation of plaque progression was mostly acute coronary syndrome. Results emphasize the need for further study to refine the methods to identify potentially vulnerable but clinically silent plaques. PMID- 18388081 TI - Age-related differences in arterial compliance are independent of body mass index. AB - The aim of this study is to examine the effect of age on large and small arterial compliance in apparently healthy adults, and to determine whether this effect is independent of body mass index. The subjects consisted of 126 men and women, aged 45 years and above. The subjects rested supine while pulse contour analysis was measured from the radial artery to evaluate large and small arterial compliance. Large (12.0 +/- 4.2 mL/mm Hg x 10, mean +/- standard deviation) and small (3.3 +/ 1.9 mL/mm Hg x 100) arterial compliance were lower in the oldest group (P = .007, P = .002, respectively) compared with the youngest group (15.2 +/- 4.6 mL/mm Hg x 10 and 5.0 +/- 2.5 mL/mm Hg x 100, respectively). After adjusting for body mass index, large and small arterial compliance remained lower in the oldest group. Large and small arterial compliance are decreased with advancing age, independent of body mass index. PMID- 18388082 TI - Relationship between collateral circulation and successful myocardial reperfusion in acute myocardial infarction: a subanalysis of the PREMIAR trial. AB - The aim of this study was to determine whether the presence of collateral circulation had a beneficial effect following primary angioplasty. In all, 114 patients who underwent primary angioplasty were included. Patients with collateral circulation had lower basal ST-segment deviation (P = .004), white cell count ( P = .001), peak creatine kinase (P = .001), and regional wall motion score values (P = .03) than patients without collateral circulation. After the procedure, the group with collaterals was associated with higher rates of normal myocardial blush, complete ST resolution, and shorter time to stable ST-recovery. Multivariable logistic analysis identified the presence of collateral circulation as independent predictor of normal myocardial blush (adjusted odds ratio = 3.98, 95% confidence interval, 1.12-14.09; P = .033) and rapid reperfusion (time to stable ST-segment recovery <7 minutes, adjusted odds ratio = 4.0, 95% confidence interval, 1.57-10.20; P = .004). The presence of collateral circulation has a protective effect on infarct size, resulting in faster reperfusion. PMID- 18388083 TI - Apical hypertrophic cardiomyopathy mimicking acute coronary syndrome: a case report and review of the literature. AB - Apical hypertrophic cardiomyopathy is a form of hypertrophic cardiomyopathy localized to the left ventricular apex. It is common in Japanese and other Asian populations, where it is generally considered relatively benign. However, its presence has also been recognized, though less commonly, in non-Asian patients. In these patients, the electrocardiographic changes and symptoms associated with apical hypertrophic cardiomyopathy often mimic acute coronary syndromes. Invasive or noninvasive evaluation of the left ventricular cavity confirms the diagnosis, with the "ace-of-spades" sign on left ventriculography being pathognomonic. Its prognosis is relatively benign in terms of cardiovascular mortality; however, morbid sequelae, such as diastolic dysfunction, left atrial enlargement, apical thrombi, ventricular aneurysms, and myocardial infarction, are not uncommon. The authors present a case of apical hypertrophic cardiomyopathy in a Caucasian patient who presented with findings suggestive of acute coronary syndrome and review the literature on apical hypertrophic cardiomyopathy. PMID- 18388085 TI - Endovascular embolization of intracranial aneurysms. AB - Intracranial aneurysm (ICA) is a common condition but with a high mortality rate when rupture occurs. The treatment of ruptured or unruptured ICA, especially with an endovascular approach, has been evolving rapidly. The current generally accepted opinion suggests that endovascular embolization is an effective technique for preventing the recurrence of aneurysm rupture, but the rebleeding rate after endovascular embolization is found to be higher than that after surgical clipping. In addition, long-term follow-up data are required for the evaluation of the effectiveness of endovascular treatment in unruptured ICA. This review presents the current understanding of ICA, the selection of optimal treatment approaches, and in particular, the advances in endovascular embolization in the treatment of ICA, including embolic materials, therapeutic and assisting techniques, long-term effectiveness, and limitations. PMID- 18388084 TI - Age and blood pressure levels modify the functional properties of central but not peripheral arteries. AB - The effect of age and blood pressure on the carotid and the radial artery distensibilities was investigated. Patients referred to the outpatient clinic of the Department of Internal Medicine and Geriatric Medicine were asked to participate in the study. The carotid and radial artery distensibility coefficients were measured. Linear regression analyses were performed to investigate the associations between determinants and arterial distensibility. The mean age of the participants was 72.3 years, and 41.5% were men. Carotid distensibility decreased with age in adjusted models (beta = -0.317; 95% confidence interval [CI], -0.241, -0.055), whereas the radial distensibility did not decrease. Levels of systolic blood pressure and mean arterial pressure were associated with decreasing levels of carotid distensibility, whereas the diastolic blood pressure and pulse pressure were not associated (beta = -0.571; 95% CI, -0.404, -0.007; beta = -0.410; 95% CI, -0.308, -0.101, respectively). In conclusion, age and blood pressure levels are associated with the distensibility of the central arteries but not with that of the peripheral arteries. PMID- 18388086 TI - Imaging investigation of a giant splenic artery aneurysm. AB - Splenic artery aneurysms represent approximately 60% of visceral arterial aneurysms. Their incidence at autopsy is 0.02 - 2 % in all age groups and rises to 10.4% above the age of 60. Although splenic artery aneurysms are the most common visceral aneurysms, giant splenic artery aneurysms >10 cm in diameter have rarely been reported. A recent review of the literature came up with 12 true giant splenic artery aneurysms >10 cm. A case of 12-cm splenic artery aneurysms is presented in this study and the imaging findings are described. PMID- 18388087 TI - Surrogate markers for atherosclerosis in overweight subjects with atherogenic dyslipidemia: the GEMS project. AB - Metabolic syndrome is a constellation of major risk factors for cardiovascular disease. In affected individuals with this syndrome, the independent contribution of low high-density lipoprotein-cholesterol and increased triglyceride levels to the development of atherosclerosis remains to be clarified. We assessed the relationship between these 2 parameters and several surrogate markers for atherosclerosis. One hundred and twenty overweight cases, defined as having high density lipoprotein-cholesterol (or=75 percentile) were compared with 120 discordant overweight controls defined on lipid values (high density lipoprotein-cholesterol >or=50 percentile and triglycerides 32 mg/L of triclosan, respectively, for inhibition. The genotype of fabI did not correlate with triclosan MIC. Artificial overexpression and mutagenesis of fabI in SL1344 each resulted in low-level triclosan resistance, indicating that FabI alone does not mediate high-level triclosan resistance in Salmonella Typhimurium. Active efflux of triclosan via AcrAB-TolC confers intrinsic resistance to triclosan as inactivation of acrB and tolC in wild-type strains and the triclosan-resistant mutants led to large decreases in triclosan resistance, which were reversed by complementation. Exemplars of each phenotype were evaluated for fitness in vivo; no fitness cost was seen and mutants colonized and persisted in chickens throughout a 28 day competitive index experiment. CONCLUSIONS: These data show that triclosan resistance can occur via distinct pathways in salmonella and that mutants selected after single exposure to triclosan are fit enough to compete with wild type strains. PMID- 18388113 TI - Severe hypokalaemia caused by flucloxacillin. PMID- 18388114 TI - Maternal occupational exposures and risk of spontaneous abortion in veterinary practice. AB - OBJECTIVES: To examine the relationship between occupational exposures and spontaneous abortion in female veterinarians. METHODS: The Health Risks of Australian Veterinarians project (HRAV) was a questionnaire-based survey of all graduates from Australian veterinary schools from 1960 to 2000. Of 5748 eligible veterinarians sent the questionnaires, 2800 replied including 1197 females (42.8%). The response rate was 59% of women veterinarians eligible to participate. The pregnancy of women was defined as the unit of analysis. We restricted analyses to pregnancies of those women who reported being employed when the pregnancy began and were working only in clinical practice. Of 1355 pregnancies, 940 were eligible for the final analysis. Self-reported occupational exposures to anaesthetic gases, x rays, pesticides and long working hours in relation to spontaneous abortion were examined. RESULTS: In a multiple logistic regression controlling for 12 potential confounders, there was a more than twofold significant increase (OR 2.49, 95% CI 1.02 to 6.04) in the risk of spontaneous abortion in women exposed to unscavenged anaesthetic gases for > or =1 h per week. Veterinarians who reported performing more than five radiographic examinations per week had a statistically significant elevated risk of spontaneous abortion compared to those who performed five or less (OR 1.82, 95% CI 1.17 to 2.82). There was also approximately a twofold significant increased risk of spontaneous abortion in women who used pesticides at work (OR 1.88, 95% CI 1.18 to 3.00). CONCLUSION: Female veterinarians, particularly those of childbearing age, should be fully informed of the possible reproductive effects of unscavenged anaesthetic gases, ionising radiation and pesticide exposure and reduce their exposure by using protective devices when they are planning to become pregnant and during pregnancy. PMID- 18388115 TI - Psychosocial work characteristics and anxiety and depressive disorders in midlife: the effects of prior psychological distress. AB - OBJECTIVES: The association between work stressors and adult psychiatric diagnoses may be biased by prior psychological distress influencing perception of work or selection into unfavourable work. This study examines the extent to which the association between work stressors and adult psychiatric diagnoses is explained by associations with earlier psychological distress and whether childhood and early adulthood psychological distress influences reported midlife work characteristics. METHODS: Follow-up at 45 years of age of 8243 participants in paid employment from the 1958 British Birth Cohort. Karasek's work characteristics and psychiatric diagnoses (Revised Clinical Interview Schedule) were measured at 45 years. Childhood internalising and externalising problems were measured at 7, 11 and 16 and malaise at 23 and 33 years. RESULTS: Internalising behaviours in childhood and early adult psychological distress predicted adverse work characteristics in mid-adulthood. High job demands (women: relative risk (RR) = 1.75, 95% CI 1.2 to 2.5; men: RR = 4.99, 95% CI 2.5 to 10.1), low decision latitude (RR = 1.46, 95% CI 1.1 to 1.9), high job strain (OR = 1.88, 95% CI 1.5 to 2.4), low work social support (RR = 1.97, 95% CI 1.5 to 2.6) and high job insecurity (OR = 1.86, 95% CI 1.4 to 2.4) were associated with mid-adulthood diagnoses. The association between work stressors and mid-adulthood diagnoses remained after adjustment for internalising behaviours and malaise at 23 and 33 years although diminished slightly in magnitude (eg, adjusted RR for support = 1.82, 95% CI 1.4 to 2.4; job strain OR = 1.78, 95% CI 1.4 to 2.3). CONCLUSIONS: Childhood and early adulthood psychological distress predict work characteristics in mid-adulthood but do not explain the associations of work characteristics with depressive episode and generalised anxiety disorder in midlife. Work stressors are an important source of preventable psychiatric diagnoses in midlife. Psychological distress may influence selection into less advantaged occupations with poorer working conditions that may increase the risk of future depressive and anxiety disorders. PMID- 18388116 TI - Pioglitazone attenuates diabetic nephropathy through an anti-inflammatory mechanism in type 2 diabetic rats. AB - BACKGROUND: Peroxisome proliferator-activated receptors (PPARs) are nuclear transcription factors that play a role in insulin sensitivity, lipid metabolism and inflammation. However, the effects of PPARgamma agonist on renal inflammation have not been fully examined in type 2 diabetic nephropathy. METHODS: In the present study, we investigated the effect and molecular mechanism of the PPARgamma agonist, pioglitazone, on the progression of diabetic nephropathy in type 2 diabetic rats. Inflammatory markers including NF-kappaB, MCP-1 and pro fibrotic cytokines were determined by RT-PCR, western blot, immunohistochemical staining and EMSA. In addition, to evaluate the direct anti-inflammatory effect of PPARgamma agonist, we performed an in vitro study using mesangial cells. RESULTS: Treatment of OLETF rats with pioglitazone improved insulin sensitivity and kidney/body weight, but had a little effect on blood pressure. Pioglitazone treatment markedly reduced urinary albumin and MCP-1 excretion, and ameliorated glomerulosclerosis. In cDNA microarray analysis using renal cortical tissues, several inflammatory and profibrotic genes were significantly down-regulated by pioglitazone including NF-kappaB, CCL2, TGFbeta1, PAI-1 and VEGF. In renal tissues, pioglitazone treatment significantly reduced macrophage infiltration and NF-kappaB activation in association with a decrease in type IV collagen, PAI-1, and TGFbeta1 expression. In cultured mesangial cells, pioglitazone-activated endogenous PPARgamma transcriptional activity and abolished high glucose-induced collagen production. In addition, pioglitazone treatment also markedly suppressed high glucose-induced MCP-1 synthesis and NF-kappaB activation. CONCLUSIONS: These data suggest that pioglitazone not only improves insulin resistance, glycaemic control and lipid profile, but also ameliorates renal injury through an anti inflammatory mechanism in type 2 diabetic rats. PMID- 18388117 TI - Metformin-associated lactic acidosis in type 2 diabetes mellitus: incidence and presentation in common clinical practice. PMID- 18388118 TI - Successful re-challenge with darbepoetin in a patient with rHu-EPO-induced pure red cell aplasia refractory to immunosuppressive drugs. PMID- 18388119 TI - A formula to predict corrected calcium in haemodialysis patients. AB - BACKGROUND: The conventional calcium correction formula (corrected total calcium (mmol/L) = TCa (mmol/L) + 0.02 [40 (g/L) - albumin (g/L)]) is broadly applied for the estimation of serum calcium in haemodialysis (HD) patients, despite the fact that it was not derived or validated in a HD population. A novel formula was derived and validated for corrected serum calcium in HD patients. METHODS: Total calcium (TCa), ionized calcium (iCa(2+)), magnesium, phosphate, albumin and bicarbonate were collected from 60 HD patients to derive the formula. A validation set of 237 stable HD patients was then examined, and subjects were classified as hyper-, hypo- and normocalcaemic based on the iCa(2+). Agreement of the new formula was calculated with iCa(2+) as the gold standard, using the intraclass correlation coefficient (ICC). This was compared to the agreement between iCa(2+) and the following: uncorrected total serum calcium (TCa), the conventional correction formula, the Orrell formula and the Clase formula. RESULTS: Using multiple linear regression the following formula was derived: corrected total calcium (mmol/L) = TCa (mmol/L) + 0.01 [30 (g/L) - albumin (g/L)]. The new formula had superior agreement compared to all of the other formulae. There was a statistically significant greater agreement between the new formula and the iCa(2+) as compared to the conventional formula (P < 0.01). However, the new formula did not significantly outperform the Orrell formula, the Clase formula or Total calcium. CONCLUSIONS: The use of our simple new formula should enable more appropriate decision making compared to the conventional formula in the highly complex HD population. PMID- 18388120 TI - S-cystatin C formulae or combination of s-cystatin C and s-creatinine formulae do not improve prediction of GFR. PMID- 18388121 TI - Coronary artery bypass surgery and acute kidney injury--impact of the off-pump technique. AB - BACKGROUND: Acute kidney injury (AKI) is a serious and frequent complication after coronary artery bypass grafting (CABG). Cardiopulmonary bypass (CPB) was identified as a major AKI risk factor after CABG. Our aim was to assess the impact of the off-pump coronary artery bypass (OPCAB) compared to the on-pump coronary artery bypass (ONCAB) technique on the rate and severity of AKI, while taking other risk factors for AKI into account. METHODS: An observational study of 201 consecutive adult patients was conducted; 100 were operated by the OPCAB and 101 by the ONCAB technique. All patients in each group were operated by a single, experienced surgeon. Fifteen pre-, intra- and postoperative variables that were repeatedly identified in previous studies as independent AKI risk factors were included in this analysis. AKI was defined as an increase of serum creatinine >/=50% or >/=0.3 mg/dL within 48 h and AKI severity was classified, according to current AKIN definitions. RESULTS: Significantly fewer OPCAB patients developed AKI compared to ONCAB (14.0 versus 27.7%; P = 0.03). OPCAB was associated with milder stages of AKI, whereas ONCAB patients had more severe AKI. Congestive heart failure and chronic kidney disease were independent risk factors for AKI. The OPCAB technique for CABG was identified as the only independent factor associated with lower incidence of AKI. CONCLUSIONS: Using current AKI definitions and classifications, the OPCAB technique for CABG, which avoids CPB; was associated with a significantly lower rate and less severe AKI compared to ONCAB. The OPCAB technique was identified as the only modifiable and potentially protective factor against postoperative AKI. PMID- 18388122 TI - Calcium load during administration of calcium carbonate or sevelamer in individuals with normal renal function. AB - BACKGROUND: Under steady-state conditions urinary calcium (Ca) excretion corresponds to the Ca load in healthy subjects. However, in chronic haemodialysis patients reliable data on Ca load are not available. In these patients Ca containing phosphate binders are suspected to play a role in the progression of arteriosclerosis via increased but not quantified Ca load. The present study evaluated the effect of calcium carbonate (CC) and sevelamer hydrochloride (SEV), a calcium-free phosphate binder, on serum Ca and urinary Ca excretion in healthy individuals. METHODS: Twelve healthy male individuals were included in a monocentre, randomized, single-blind, placebo-controlled, three-way crossover phase I study. Concurrently with their meals, participants received 4 x 2 tablets of SEV (800 mg), CC (500 mg) or placebo for 6 days with 1-week washout between the treatment periods. During the study, weekly blood samples were taken and 24-h urine was collected each day for measurement of calcium, magnesium, phosphorus, chloride and iPTH. RESULTS: Mean daily urinary phosphorus excretion was significantly lower in subjects undergoing SEV treatment compared to those taking placebo (P < 0.001). Mean daily total urinary excretion of calcium was significantly higher in CC-treated participants compared to those receiving placebo (P < 0.001). Mean 24-h calcium excretion during the 6 treatment days was 6.60 +/- 2.62 mmol [265 +/- 105 mg] (CC) versus 5.15 +/- 2.16 mmol [206 +/- 87 mg] (SEV) versus 4.95 +/- 1.63 mmol [198 +/- 65 mg] (Placebo). Taking into account nutritional calcium intake estimated from dietary records fractional calcium absorption was 8.7% (CC), 13.3% (placebo) and 14.8% (sevelamer). CONCLUSION: Intake of calcium carbonate compared to placebo in contrast to sevelamer in healthy individuals was associated with increased total urinary calcium excretion indicating an increased calcium load due to increased intestinal calcium absorption. PMID- 18388123 TI - Free water transport, small pore transport and the osmotic pressure gradient three-pore model of peritoneal transport. PMID- 18388124 TI - Impact of subclinical carotid atherosclerosis on incident chronic kidney disease in the elderly. AB - BACKGROUND: Cardiovascular disease (CVD) is increased in persons with chronic kidney disease (CKD); however, no prospective studies have examined carotid intima-media thickness (CIMT) as a risk factor for CKD. METHODS: A total of 2751 participants who were in the Intervention Project on Cerebrovascular Diseases and Dementia in the community of Ebersberg, Bavaria study and had normal baseline kidney function composed the study cohort. Measures of kidney function were estimated glomerular filtration rate (eGFR) using the Cockcroft-Gault formula in mL/min/1.73 m(2). The main outcome measure was incident CKD defined as an eGFR < 60 mL/min/1.73 m(2) at study year 2 among those with an eGFR >or= 60 mL/min/1.73 m(2) at baseline. Multivariate Cox regression models were used to assess the association between CIMT and incident CKD. RESULTS: In multivariate analysis, there was a significant and graded association in eGFR decline, -8 +/- 21, -10 +/ 22, -11 +/- 12 and -15 +/- 11 mL/min/1.73 m(2) for the baseline CIMT quartiles 1 (<0.66 mm), 2 (0.66-0.77 mm), 3 (0.77-0.88 mm) and 4 (>0.88 mm), respectively (P for trend: 0.01), during the 2-year follow-up period. Subjects with a baseline CIMT in the fourth quartile developed a significant decrease in eGFR as compared with subjects with a baseline CIMT in the first quartile (P < 0.01). After adjustment for confounding factors, the baseline CIMT remained a predictor for the occurrence of CKD [hazard ratio, 1.17; 95% confidence interval (CI), 1.08 1.30]. CONCLUSIONS: Increases in CIMT, as measured non-invasively by ultrasonography, are directly associated with an increased risk of CKD in elderly individuals. PMID- 18388125 TI - Identification of the coiled-coil domains of Enterococcus faecalis DivIVA that mediate oligomerization and their importance for biological function. AB - Bacillus subtilis (Bs) DivIVA comprises coiled-coil structures and self associates forming a 10-12 mer complex in vitro. Using bioinformatic approaches, we determined that Enterococcus faecalis (Ef) DivIVA comprises four coiled-coil domains, one at the N-terminus, the second and the third in the central region of the protein and the fourth at the C-terminus. We determined that DivIVA(Ef) self interacts and forms a 10-12 multimeric complex. Point mutations or deletions of the central regions predicted bioinformatically to disrupt the coiled-coil structures either eliminated or weakened DivIVA(Ef) self-interaction and reduced oligomerization. Mutations disrupting the N- and C-terminal coiled-coils of DivIVA(Ef) did not affect DivIVA(Ef) oligomerization. The introduction of DivIVA(Ef) mutations to both the N-terminal and the central coiled-coil domains were lethal unless rescued by expressing wild-type DivIVA(Ef) in trans. E. faecalis cells expressing these mutations displayed aberrant cell morphology, indicating disruption of the normal cell division phenotype. The results in E. faecalis also indicate that both the N-terminal and the central coiled-coil structures of DivIVA(Ef) are indispensable for proper biological function. Overexpression of wild-type DivIVA(Ef) in both rod-shaped and round Escherichia coli cells resulted in morphological changes, while the overexpression of DivIVA(Ef) mutations failed to induce such alterations. PMID- 18388127 TI - Phosphoproteomic analysis of the mouse brain cytosol reveals a predominance of protein phosphorylation in regions of intrinsic sequence disorder. AB - We analyzed the mouse forebrain cytosolic phosphoproteome using sequential (protein and peptide) IMAC purifications, enzymatic dephosphorylation, and targeted tandem mass spectrometry analysis strategies. In total, using complementary phosphoenrichment and LC-MS/MS strategies, 512 phosphorylation sites on 540 non-redundant phosphopeptides from 162 cytosolic phosphoproteins were characterized. Analysis of protein domains and amino acid sequence composition of this data set of cytosolic phosphoproteins revealed that it is significantly enriched in intrinsic sequence disorder, and this enrichment is associated with both cellular location and phosphorylation status. The majority of phosphorylation sites found by MS were located outside of structural protein domains (97%) but were mostly located in regions of intrinsic sequence disorder (86%). 368 phosphorylation sites were located in long regions of disorder (over 40 amino acids long), and 94% of proteins contained at least one such long region of disorder. In addition, we found that 58 phosphorylation sites in this data set occur in 14-3-3 binding consensus motifs, linear motifs that are associated with unstructured regions in proteins. These results demonstrate that in this data set protein phosphorylation is significantly depleted in protein domains and significantly enriched in disordered protein sequences and that enrichment of intrinsic sequence disorder may be a common feature of phosphoproteomes. This supports the hypothesis that disordered regions in proteins allow kinases, phosphatases, and phosphorylation-dependent binding proteins to gain access to target sequences to regulate local protein conformation and activity. PMID- 18388126 TI - Use of an immunoaffinity-mass spectrometry-based approach for the quantification of protein biomarkers from serum samples of lung cancer patients. AB - It is a challenging task to verify and quantify potential biomarkers expressed at elevated levels in sera from cancer patients. An immunoaffinity-mass spectrometry based approach has been developed using antibodies to enrich proteins of interest from sera followed by mass spectrometry-based quantification. Antibodies specific to the protein of interest were immobilized to hydrazide resin via the carbohydrate moiety on the Fc region of the antibody. Captured proteins were eluted, reduced, alkylated, and digested with trypsin. Peptides were analyzed by LC coupled with multiple reaction monitoring approach, and quantification was achieved by the addition of stable isotope-labeled (heavy) standard peptides. Using this methodology, we were able to achieve a linear response from 15 to 250 ng/ml for carcinoembryonic antigen (CEA), a known tumor biomarker. Moreover we observed elevated levels of CEA in sera samples from lung cancer patients that to our knowledge is the first time that circulating CEA has been detected by mass spectrometry-based analysis. This approach was further applied to potential protein biomarkers discovered from tumor cell lines and tumor tissues. A linear response was obtained from a multiplex spiking experiment in normal human sera for secretory leukocyte peptidase inhibitor (4-500 ng/ml), tissue factor pathway inhibitor (TFPI) (42-1000 ng/ml), tissue factor pathway inhibitor 2 (TFPI2) (2 250 ng/ml), and metalloproteinase inhibitor 1 (TIMP1) (430-1000 ng/ml). A replicate experiment for a single concentration value yielded a relative coefficient of variation better than 11% for TFPI, secretory leukocyte peptidase inhibitor, and TFPI2. The expression level of the proteins in lung cancer patient sera was assayed by an immunoaffinity-multiple reaction monitoring method, and the results were comparable with those obtained from ELISA. This immunoaffinity mass spectrometry-based quantification approach thus provides a specific and accurate assay for verifying the expression of potential biomarkers in patient serum samples especially for those proteins for which the necessary reagents for ELISA development are unavailable. PMID- 18388128 TI - The merest Logomachy: The 1868 Norwich discussion of aphasia by Hughlings Jackson and Broca. AB - This article reconsiders the events that took place at the 1868 meeting of the British Association for the Advancement of Science (BA) in Norwich. Paul Broca and John Hughlings Jackson were invited to speak on the new and controversial subject of aphasia. Over the ensuing decades, there have been repeated references made to a debate between Broca and Jackson. This meeting has been identified as a turning point in favour of Broca's position on the cerebral localization of language. A return to original sources from key witnesses reveals that the opinion of the British practitioners was generally against Broca's views. Close examination of contemporaneous materials suggests that no public debate between Jackson and Broca occurred. However, the public discussion after Broca's presentation records notable concerns over both theoretical issues of localization of function and the status of exceptional clinical cases. A significant stage in the development of current views on the organization of language in the brain is revealed in the accounts of the BA meeting in August 1868 and successive responses to these events in the British press over a period of years. PMID- 18388129 TI - The poly(A) site sequence in HDV RNA alters both extent and rate of self-cleavage of the antigenomic ribozyme. AB - The ribozyme self-cleavage site in the antigenomic sequence of hepatitis delta virus (HDV) RNA is 33-nt downstream of the poly(A) site for the delta antigen mRNA. An HDV antigenomic ribozyme precursor RNA that included the upstream poly(A) processing site was used to test the hypothesis that nonribozyme sequence near the poly(A) site could affect ribozyme activity. Relative to ribozyme precursor without the extra upstream sequences, the kinetic profile for self cleavage of the longer precursor was altered in two ways. First, only half of the precursor RNA self-cleaved. The cleaved fraction could be increased or decreased with mutations in the upstream sequence. These mutations, which were predicted to alter the relative stability of competing secondary structures within the precursor, changed the distribution of alternative RNA structures that are resolved in native-gel electrophoresis. Second, the active fraction cleaved with an observed rate constant that was higher than that of the ribozyme without the upstream sequences. Moreover, the higher rate constants occurred at lower, near physiological, divalent metal ion concentrations (1-2 mM). Modulation of ribozyme activity, through competing alternative structures, could be part of a mechanism that allows a biologically significant choice between maturation of the mRNA and processing of replication intermediates. PMID- 18388130 TI - Formation of the double helix: a mutational study. AB - To investigate the mechanisms by which oligonucleotides hybridize to target molecules, the binding of two oligodeoxynucleotide probes to RNA targets was measured over a broad range of temperatures. Mutations were then scanned across each DNA/RNA hybrid to map, at single base resolution, sequences important for hybridization. Despite being unrelated in sequence, each hybrid formed by a similar mechanism. In the absence of secondary structure, two stretches of bases, termed nucleation regions, cooperated with one another by a looping mechanism to nucleate hybridization. Mutations inside each nucleation region strongly decreased hybridization rates, even at temperatures well below the melting temperature (T(m)) of the hybridized duplex. Surprisingly, nucleation regions were detected in a RNA target but not a corresponding DNA target. When either nucleation region was sequestered in secondary structure, the hybridization rate fell and the mechanism of hybridization changed. Single-stranded bases within the nucleation region of the probe and target first collided to form a double helix. If sufficiently G + C rich, the double helix then propagated throughout the oligonucleotide by a strand invasion process. On the basis of these results, general mechanisms for the hybridization of oligonucleotides to complementary and mutant targets are proposed. PMID- 18388131 TI - Transcriptional activation of the tumor suppressor and differentiation gene S100A2 by a novel p63-binding site. AB - S100A2 is generally found expressed in the epidermis and was recently shown to play a crucial role in the differentiation of keratinocytes. Also known as CaN19, S100A2 was identified as a potential tumor suppressor. Expression of S100A2 is upregulated by p53. The proteins p63 and p73 are related to p53 and are expressed as several splice variants with partially overlapping tasks but also functions different from p53. It had been shown that p63 proteins with mutations in their DNA-binding domain cause severe phenotypes in man as autosomal dominantly inherited disease including EEC, AEC, SHFM, LMS and ADULT syndromes. Here we show that S100A2 is a transcriptional target of p63/p73 family members, particularly the p63 splice variant TAp63gamma. The regulation is mediated by a novel transcriptional element in the S100A2 promoter which is bound by TAp63gamma but not by p53. Mutant p63 proteins derived from EEC and ADULT syndrome patients cannot activate S100A2 transcription whereas SHFM-related mutants still can stimulate the S100A2 promoter. Consistent with a function in tumor suppression S100A2 expression is stimulated upon DNA damage. After doxorubicin treatment p63gamma proteins are recruited to the S100A2 promoter in vivo. This may indicate a function of the p63-dependent S100A2 regulation in tumor suppression. PMID- 18388132 TI - A shared RNA-binding site in the Pet54 protein is required for translational activation and group I intron splicing in yeast mitochondria. AB - The Pet54p protein is an archetypical example of a dual functioning ('moonlighting') protein: it is required for translational activation of the COX3 mRNA and splicing of the aI5beta group I intron in the COX1 pre-mRNA in Saccharomyces cerevisiae mitochondria (mt). Genetic and biochemical analyses in yeast are consistent with Pet54p forming a complex with other translational activators that, in an unknown way, associates with the 5' untranslated leader (UTL) of COX3 mRNA. Likewise, genetic analysis suggests that Pet54p along with another distinct set of proteins facilitate splicing of the aI5beta intron, but the function of Pet54 is, also, obscure. In particular, it remains unknown whether Pet54p is a primary RNA-binding protein that specifically recognizes the 5' UTL and intron RNAs or whether its functional specificity is governed in other ways. Using recombinant protein, we show that Pet54p binds with high specificity and affinity to the aI5beta intron and facilitates exon ligation in vitro. In addition, Pet54p binds with similar affinity to the COX3 5' UTL RNA. Competition experiments show that the COX3 5'UTL and aI5beta intron RNAs bind to the same or overlapping surface on Pet54p. Delineation of the Pet54p-binding sites by RNA deletions and RNase footprinting show that Pet54p binds across a similar length sequence in both RNAs. Alignment of the sequences shows significant (56%) similarity and overlap between the binding sites. Given that its role in splicing is likely an acquired function, these data support a model in which Pet54p's splicing function may have resulted from a fortuitous association with the aI5beta intron. This association may have lead to the selection of Pet54p variants that increased the efficiency of aI5beta splicing and provided a possible means to coregulate COX1 and COX3 expression. PMID- 18388133 TI - Community health workers and the response to HIV/AIDS in South Africa: tensions and prospects. AB - After a decline in enthusiasm for national community health worker (CHW) programmes in the 1980s, these have re-emerged globally, particularly in the context of HIV. This paper examines the case of South Africa, where there has been rapid growth of a range of lay workers (home-based carers, lay counsellors, DOT supporters etc.) principally in response to an expansion in budgets and programmes for HIV, most recently the rollout of antiretroviral therapy (ART). In 2004, the term community health worker was introduced as the umbrella concept for all the community/lay workers in the health sector, and a national CHW Policy Framework was adopted. We summarize the key features of the emerging national CHW programme in South Africa, which include amongst others, their integration into a national public works programme and the use of non-governmental organizations as intermediaries. We then report on experiences in one Province, Free State. Over a period of 2 years (2004--06), we made serial visits on three occasions to the first 16 primary health care facilities in this Province providing comprehensive HIV services, including ART. At each of these visits, we did inventories of CHW numbers and training, and on two occasions conducted facility-based group interviews with CHWs (involving a total of 231 and 182 participants, respectively). We also interviewed clinic nurses tasked with supervising CHWs. From this evaluation we concluded that there is a significant CHW presence in the South African health system. This infrastructure, however, shares many of the managerial challenges (stability, recognition, volunteer vs. worker, relationships with professionals) associated with previous national CHW programmes, and we discuss prospects for sustainability in the light of the new policy context. PMID- 18388134 TI - Coincidence detection of convergent perforant path and mossy fibre inputs by CA3 interneurons. AB - We performed whole-cell recordings from CA3 s. radiatum (R) and s. lacunosum moleculare (L-M) interneurons in hippocampal slices to examine the temporal aspects of summation of converging perforant path (PP) and mossy fibre (MF) inputs. PP EPSPs were evoked from the s. lacunosum-moleculare in area CA1. MF EPSPs were evoked from the medial extent of the suprapyramidal blade of the dentate gyrus. Summation was strongly supralinear when examining PP EPSP with MF EPSP in a heterosynaptic pair at the 10 ms ISI, and linear to sublinear at longer ISIs. This pattern of nonlinearities suggests that R and L-M interneurons act as coincidence detectors for input from PP and MF. Summation at all ISIs was linear in voltage clamp mode demonstrating that nonlinearities were generated by postsynaptic voltage-dependent conductances. Supralinearity was not detected when the first EPSP in the pair was replaced by a simulated EPSP injected into the soma, suggesting that the conductances underlying the EPSP boosting were located in distal dendrites. Supralinearity was selectively eliminated with either Ni2+ (30 microm), mibefradil (10 microm) or nimodipine (15 microm), but was unaffected by QX-314. This pharmacological profile indicates that supralinearity is due to recruitment of dendritic T-type Ca2+channels by the first subthreshold EPSP in the pair. Results with the hyperpolarization-activated (Ih) channel blocker ZD 7288 (50 microm) revealed that Ih restricted the time course of supralinearity for coincidently summed EPSPs, and promoted linear to sublinear summation for asynchronous EPSPs. We conclude that coincidence detection results from the counterbalanced activation of T-type Ca2+ channels and inactivation of Ih. PMID- 18388135 TI - Reduced vascular tetrahydrobiopterin (BH4) and endothelial function with ageing: is it time for a chronic BH4 supplementation trial in middle-aged and older adults? PMID- 18388136 TI - Cytoplasmic calcium oscillations and store-operated calcium influx. AB - Intracellular calcium oscillations have fascinated scientists for decades. They provide an important cellular signal which, unlike most signalling mechanisms, is digitally encoded. While it is generally agreed that oscillations most frequently arise from cyclical release and re-uptake of intracellularly stored calcium, it is becoming increasingly clear that influx of calcium across the plasma membrane also plays a critical role in their maintenance and even in delivering their signal to the correct cellular locus. In this review we will discuss the role played by Ca(2+) entry mechanisms in Ca(2+) oscillations, and approaches to understanding the molecular nature of this Ca(2+) entry pathway. PMID- 18388138 TI - Melatonin potentiates rod signals to ON type bipolar cells in fish retina. AB - Melatonin is involved in regulation of a variety of physiological functions through activation of specific G-protein coupled receptors. However, the neuromodulatory role of melatonin, released from photoreceptors in the retina, is poorly understood. Here we show that melatonin enhances the sensitivity of the rod signal pathway by potentiating signal transfer from rod photoreceptors to ON bipolar cells (Rod-ON-BCs). Whole-cell patch-clamp recordings showed that melatonin induced a sustained inward current from Rod-ON-BCs, through activation of the melatonin MT2 receptor, which was identified as one mediated by a cGMP dependent cation channel. Consistent with this, melatonin was found, using immunocytochemistry, to increase intracellular cGMP levels, which was identified due to an inhibition of phosphodiesterase. Physiologically, melatonin potentiated responses of Rod-ON-BCs to simulated light flashes (brief puffs of CPPG, an mGluR6 antagonist, in the presence of l-AP4, an mGluR6 agonist), which was mediated by cGMP-dependent kinase, and increased the amplitude of the scotopic electroretinographic b-wave, a reflection of Rod-ON-BC activity. These results suggest that melatonin, being at a higher level at night, may improve the signal/noise ratio for rod signals in the outer retina by enhancing signal transfer from rods to BCs. PMID- 18388137 TI - Fluid flow induces mechanosensitive ATP release, calcium signalling and Cl- transport in biliary epithelial cells through a PKCzeta-dependent pathway. AB - ATP in bile is a potent secretogogue, stimulating cholangiocyte Cl- and fluid secretion via binding to membrane P2 receptors, though the physiological stimuli involved in biliary ATP release are unknown. The goal of the present studies was to determine the potential role of fluid flow in biliary ATP release and secretion. In both human Mz-Cha-1 biliary cells and normal rat cholangiocyte monolayers, exposure to flow increased relative ATP release which was proportional to the shear stress. In parallel studies, shear was associated with an increase in [Ca2+]i and membrane Cl- permeability, which were both dependent on extracellular ATP and P2 receptor stimulation. Flow-stimulated ATP release was dependent on [Ca2+]i, exhibited desensitization with repetitive stimulation, and was regulated by PKCzeta. In conclusion, both human and rat biliary cells exhibit flow-stimulated, PKCzeta-dependent, ATP release, increases in [Ca2+]i and Cl- secretion. The finding that fluid flow can regulate membrane transport suggests that mechanosensitive ATP release may be a key regulator of biliary secretion and an important target to modulate bile flow in the treatment of cholestatic liver diseases. PMID- 18388139 TI - Preoptic mechanism for cold-defensive responses to skin cooling. AB - We recently identified a somatosensory pathway that transmits temperature information from the skin to a median subregion of the preoptic area (POA), a thermoregulatory centre. Here, we investigated in vivo the local neuronal circuit in the rat POA that processes the thermosensory information and outputs thermoregulatory effector signals. Skin cooling-evoked increases in sympathetic thermogenesis in brown adipose tissue, in metabolism and in heart rate were reversed by inhibition of neurons in the median preoptic nucleus (MnPO). Glutamatergic stimulation or disinhibition of MnPO neurons evoked thermogenic, metabolic and cardiac responses that mimicked the cold-defensive responses to skin cooling and were reversed by antagonizing GABA(A) receptors in the medial preoptic area (MPO), which is thought to contain neurons providing thermoregulatory output to effectors. These results suggest that GABA inhibition of output neurons in the MPO by MnPO neurons that are activated by cool sensory signals from the skin is a core thermoregulatory mechanism within the POA that is essential for the feedforward defence of body temperature against cold challenges in the environment. PMID- 18388141 TI - GeneTrack--a genomic data processing and visualization framework. AB - MOTIVATION: High-throughput 'ChIP-chip' and 'ChIP-seq' methodologies generate sufficiently large data sets that analysis poses significant informatics challenges, particularly for research groups with modest computational support. To address this challenge, we devised a software platform for storing, analyzing and visualizing high resolution genome-wide binding data. GeneTrack automates several steps of a typical data processing pipeline, including smoothing and peak detection, and facilitates dissemination of the results via the web. Our software is freely available via the Google Project Hosting environment at http://genetrack.googlecode.com PMID- 18388142 TI - MTMDAT: Automated analysis and visualization of mass spectrometry data for tertiary and quaternary structure probing of proteins. AB - In structural biology and -genomics, nuclear magnetic resonance (NMR) spectroscopy and crystallography are the methods of choice, but sample requirements can be hard to fulfil. Valuable structural information can also be obtained by using a combination of limited proteolysis and mass spectrometry, providing not only knowledge of how to improve sample conditions for crystallization trials or NMR spectrosopy by gaining insight into subdomain identities but also probing tertiary and quaternary structure, folding and stability, ligand binding, protein interactions and the location of post translational modifications. For high-throughput studies and larger proteins, however, this experimentally fast and easy approach produces considerable amounts of data, which until now has made the evaluation exceedingly laborious if at all manually possible. MTMDAT, equipped with a browser-like graphical user interface, accelerates this evaluation manifold by automated peak picking, assignment, data processing and visualization. AVAILABILITY: MTMDAT can be downloaded from the following page: http://www.cms.liu.se/chemistry/molbiotech/maria_sunnerhagens_group/mtmdat by clicking on the corresponding links (windows- or unix-based) together with the manual and example files. The program is free for academic/non-commercial purposes only. PMID- 18388143 TI - Prediction of the translocon-mediated membrane insertion free energies of protein sequences. AB - MOTIVATION: Helical membrane proteins (HMPs) play crucial roles in a variety of cellular processes. Unlike water-soluble proteins, HMPs need not only to fold but also get inserted into the membrane to be fully functional. This process of membrane insertion is mediated by the translocon complex. Thus, it is of great interest to develop computational methods for predicting the translocon-mediated membrane insertion free energies of protein sequences. RESULT: We have developed Membrane Insertion (MINS), a novel sequence-based computational method for predicting the membrane insertion free energies of protein sequences. A benchmark test gives a correlation coefficient of 0.74 between predicted and observed free energies for 357 known cases, which corresponds to a mean unsigned error of 0.41 kcal/mol. These results are significantly better than those obtained by traditional hydropathy analysis. Moreover, the ability of MINS to reasonably predict membrane insertion free energies of protein sequences allows for effective identification of transmembrane (TM) segments. Subsequently, MINS was applied to predict the membrane insertion free energies of 316 TM segments found in known structures. An in-depth analysis of the predicted free energies reveals a number of interesting findings about the biogenesis and structural stability of HMPs. AVAILABILITY: A web server for MINS is available at http://service.bioinformatik.uni-saarland.de/mins PMID- 18388140 TI - Contribution of voltage-gated sodium channels to the b-wave of the mammalian flash electroretinogram. AB - Voltage-gated sodium channels (Na(v) channels) in retinal neurons are known to contribute to the mammalian flash electroretinogram (ERG) via activity of third order retinal neurons, i.e. amacrine and ganglion cells. This study investigated the effects of tetrodotoxin (TTX) blockade of Na(v) channels on the b-wave, an ERG wave that originates mainly from activity of second-order retinal neurons. ERGs were recorded from anaesthetized Brown Norway rats in response to brief full field flashes presented over a range of stimulus energies, under dark-adapted conditions and in the presence of steady mesopic and photopic backgrounds. Recordings were made before and after intravitreal injection of TTX (approximately 3 microm) alone, 3-6 weeks after optic nerve transection (ONTx) to induce ganglion cell degeneration, or in combination with an ionotropic glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 200 microm) to block light-evoked activity of inner retinal, horizontal and OFF bipolar cells, or with the glutamate agonist N-methyl-D-aspartate (NMDA, 100-200 microm) to reduce light-evoked inner retinal activity. TTX reduced ERG amplitudes measured at fixed times corresponding to b-wave time to peak. Effects of TTX were seen under all background conditions, but were greatest for mesopic backgrounds. In dark-adapted retina, b-wave amplitudes were reduced only when very low stimulus energies affecting the inner retina, or very high stimulus energies were used. Loss of ganglion cells following ONTx did not affect b-wave amplitudes, and injection of TTX in eyes with ONTx reduced b-wave amplitudes by the same amount for each background condition as occurred when ganglion cells were intact, thereby eliminating a ganglion cell role in the TTX effects. Isolation of cone driven responses by presenting test flashes after cessation of a rod-saturating conditioning flash indicated that the TTX effects were primarily on cone circuits contributing to the mixed rod-cone ERG. NMDA significantly reduced only the additional effects of TTX on the mixed rod-cone ERG observed under mesopic conditions, implicating inner retinal involvement in those effects. After pharmacological blockade with CNQX, TTX still reduced b-wave amplitudes in cone isolated ERGs indicating Na(v) channels in ON cone bipolar cells themselves augment b-wave amplitude and sensitivity. This augmentation was largest under dark-adapted conditions, and decreased with increasing background illumination, indicating effects of background illumination on Na(v) channel function. These findings indicate that activation of Na(v) channels in ON cone bipolar cells affects the b-wave of the rat ERG and must be considered when analysing results of ERG studies of retinal function. PMID- 18388144 TI - ASPicDB: a database resource for alternative splicing analysis. AB - MOTIVATION: Alternative splicing has recently emerged as a key mechanism responsible for the expansion of transcriptome and proteome complexity in human and other organisms. Although several online resources devoted to alternative splicing analysis are available they may suffer from limitations related both to the computational methodologies adopted and to the extent of the annotations they provide that prevent the full exploitation of the available data. Furthermore, current resources provide limited query and download facilities. RESULTS: ASPicDB is a database designed to provide access to reliable annotations of the alternative splicing pattern of human genes and to the functional annotation of predicted splicing isoforms. Splice-site detection and full-length transcript modeling have been carried out by a genome-wide application of the ASPic algorithm, based on the multiple alignments of gene-related transcripts (typically a Unigene cluster) to the genomic sequence, a strategy that greatly improves prediction accuracy compared to methods based on independent and progressive alignments. Enhanced query and download facilities for annotations and sequences allow users to select and extract specific sets of data related to genes, transcripts and introns fulfilling a combination of user-defined criteria. Several tabular and graphical views of the results are presented, providing a comprehensive assessment of the functional implication of alternative splicing in the gene set under investigation. ASPicDB, which is regularly updated on a monthly basis, also includes information on tissue-specific splicing patterns of normal and cancer cells, based on available EST sequences and their library source annotation. AVAILABILITY: www.caspur.it/ASPicDB PMID- 18388145 TI - Autoinflammatory diseases: an update of clinical and genetic aspects. AB - To review clinical manifestations and genetic features of the autoinflammatory diseases, a group of rare, genetically defined diseases which have been newly grouped into a coherent whole. We performed a literature review using the keywords 'periodic fever syndrome', 'autoinflammatory disease' and 'therapy'. All relevant original and review articles in English were reviewed. A case report of each autoinflammatory disease was excerpted from the literature and presented. This review summarizes the clinical manifestations, genetic analysis and therapy of FMF, TNF-alpha receptor-associated periodic fever syndrome, hyperimmunoglobulinaemia D periodic fever syndrome and cryopyrin-associated periodic fever syndrome. These diseases have periodic fever, are hereditary and recurrent, with elevated acute-phase reactants. Differentiating features of these disorders are tabulated. Autoinflammatory diseases have some communalities in their presentation although they represent a relatively diverse group of genetically associated diseases. PMID- 18388146 TI - Altered frequency and migration capacity of CD4+CD25+ regulatory T cells in systemic lupus erythematosus. AB - OBJECTIVES: To determine the frequency and chemokine receptor-related migratory capacity of CD4(+)CD25(+) regulatory T cells (Tregs) and their association with clinical parameters in patients with SLE. METHODS: The expression of CD4, CD25, FoxP3 and CCR4 was examined with flow cytometry after staining with fluorescence conjugated antibodies in 20 patients with SLE, 20 patients with RA and 21 age- and sex-matched healthy controls. For analysis of migration capacity in 24-well chemotaxis chambers, sorted cells were stimulated with ligands of CCR4, CCL17 and CCL22 and analysed with FACScan. Correlations between the number of Tregs and CCR4(+) Treg cells and clinical parameters were analysed. RESULTS: The numbers of Tregs(bright) and CCR4(+) Tregs(bright) were significantly decreased in the patients with SLE compared with healthy controls. The number of Tregs(bright) was negatively correlated with the levels of anti-dsDNA antibody and the number of CCR4(+) Tregs(bright) had a positive correlation with the levels of C(3). Percentage of migrated Tregs(bright) by CCL17 or CCL22 was significantly decreased in the patients with SLE compared with healthy controls. CONCLUSIONS: These results suggest that altered frequency of Tregs and CCR4(+) Tregs(bright) and decreased migratory capacity of Tregs might be involved in the pathogenesis of SLE and indicate that targeting the Tregs can be a new therapeutic strategy in SLE. PMID- 18388147 TI - A new paradigm for musculoskeletal clinical trials in the UK: the Arthritis Research Campaign (ARC) Clinical Studies Groups initiative. AB - In October 2007, the UK Arthritis Research Campaign (ARC) launched a new approach for the support of clinical trials and related research in the UK. The initiative depends on a partnership between ARC, the UK Clinical Research Network (UKCRN) and the pharmaceutical and related industry. The aim is to develop nationally agreed strategic plans for intervention research for the major musculoskeletal disorders. These will range from testing experimental therapies to novel approaches/ways of using existing interventions, taking advantage of the opportunities afforded for the enhanced support for clinical trials promised by the establishment of local research networks within the National Institute for Health Research (NIHR) Comprehensive Clinical Research Network (CCRN). The initiative encourages greater collaboration with industry with a move to enhance industrial support for research strategies prioritized by the key stakeholders of health care professionals and patients. PMID- 18388148 TI - Messenger ribonucleic acid expression profile in peripheral blood cells from RA patients following treatment with an anti-TNF-alpha monoclonal antibody, infliximab. AB - OBJECTIVES: We monitored the mRNA expression profiles of peripheral blood cells during treatment with a TNF-alpha inhibitor, infliximab, in patients with RA. Using a DNA microarray analysis, we demonstrated a unique set of genes, with distinct baseline and post-treatment changes in expression between responders and non-responders to infliximab treatment. METHODS: Using a customized low-density cDNA microarray with 747 genes and a reliable data collection system, we monitored the mRNA expression profiles of whole blood cells from 18 RA patients before and after the infusion of infliximab for up to 22 weeks. The clinical response to treatment with infliximab was determined using the ACR response criteria, the disease activity score of 28 joints (DAS28), and individual clinical parameters. The patients were classified as responders or non-responders based on their ACR50% response at 22 weeks. RESULTS: Approximately 15% of the total genes were found to exhibit a >1.5-fold change, compared with their reference values, at one or more time points during the 22 weeks of infliximab therapy. The expression of inflammatory genes, such as IFN-related genes, was strongly correlated with the serum level of CRP and the DAS28. The increased expression of inflammatory genes in responders was normalized within 2 weeks and then remained at a normal level during the treatment period. In contrast, in the non-responders, the elevated expression at baseline, although it was significantly decreased at 2 weeks, returned to the baseline level after 14 weeks. In addition to inflammatory genes, we identified several groups of genes with distinct differences in expression between the responders and non responders. CONCLUSIONS: Our results suggest that a customized low-density microarray is useful for monitoring mRNA expression profiles in peripheral blood cells, enabling us to identify a unique set of genes with differentially regulated expressions in responders and non-responders to a TNF inhibitor among patients with RA. PMID- 18388149 TI - Developmental dependence on NurRE and EboxNeuro for expression of pituitary proopiomelanocortin. AB - Cell-specific expression of the pituitary proopiomelanocortin (POMC) gene depends on the combinatorial action of a large number of DNA-binding transcription factors (TFs). These include general and cell-restricted factors, as well as factors that act as effectors of signaling pathways. We have previously defined in the distal POMC promoter a composite regulatory element that contains targets for basic helix-loop-helix TFs conferring cell specificity and for NGFI-B orphan nuclear receptors that are responsive to CRH signaling and to glucocorticoid negative feedback. These factors act on neighboring regulatory elements, the Ebox(Neuro) and NurRE, respectively. Currently, the Ebox(Neuro) is thought to be the target of NeuroD1 during fetal development, but this factor may not account for activity in the adult pituitary; it is also unknown whether the NurRE and NGFI-B-related factors are active before establishment of the hypothalamic pituitary portal system. In order to assess the importance of these regulatory elements and their cognate TFs throughout pituitary organogenesis and in the adult, we have assessed the activity of mutant POMC promoters in transgenic mice throughout development. These experiments indicate that the Ebox(Neuro) and cognate basic helix-loop-helix factors are required throughout development and in the adult gland, beyond expression of NeuroD1. Similarly, the data reveal sustained importance of the NurRE and its cognate factors throughout pituitary development. These data contrast the sustained dependence throughout development on the same regulatory elements with the highly dynamic patterns of TF expression and the modulation of their activity in response to signaling pathways. PMID- 18388151 TI - Commentary: Personality and health inequality: inconclusive evidence for an indirect hypothesis. PMID- 18388150 TI - Estrogen receptor-alpha hinge-region lysines 302 and 303 regulate receptor degradation by the proteasome. AB - Cellular levels of estrogen receptor-alpha (ERalpha) protein are regulated primarily by the ubiquitin-proteasome pathway. Dynamic interactions between ERalpha and the protein degradation machinery facilitate the down-regulation process by targeting receptor lysine residues for polyubiquitination. To date, the lysines that control receptor degradation have not been identified. Two receptor lysines, K302 and K303, located in the hinge-region of ERalpha, serve multiple regulatory functions, and we examined whether these might also regulate receptor polyubiquitination, turnover, and receptor-protein interactions. We used ERalpha-negative breast cancer C4-12 cells to generate cells stably expressing wild-type (wt)ERalpha or ERalpha with lysine-to-alanine substitutions at K302 and K303 (ERalpha-AA). In the unliganded state, ERalpha-AA displayed rapid polyubiquitination and enhanced basal turnover, as compared with wtERalpha, due to its elevated association with the ubiquitin ligase carboxy terminus of Hsc70 interacting protein (CHIP) and the proteasome-associated cochaperone Bag1. Treatment of C4-12 cells with either 17beta-estradiol (E2) or the pure antiestrogen ICI 182,780 (ICI) induced rapid degradation of wtERalpha via the ubiquitin-proteasome pathway; however, in the presence of these ligands, ERalpha AA was less efficiently degraded. Furthermore, ERalpha-AA was resistant to ICI induced polyubiquitination, suggesting that these lysines are polyubiquitinated in response to the antiestrogen and demonstrate a novel role for these two lysines in the mechanism of action of ICI-induced receptor down-regulation. The reduced stability of ERalpha-AA in the unliganded state and the increased stability of ERalpha-AA in the liganded state were concordant with reporter gene assays demonstrating that ERalpha-AA has lower basal activity but higher E2 inducibility than wtERalpha. These data provide the first evidence that K302/303 protect ERalpha from basal degradation and are necessary for efficient E2- and ICI-induced turnover in breast cancer cells. PMID- 18388152 TI - Body mass index, weight change and mortality risk in a prospective study in India. AB - BACKGROUND: Although the detrimental effect of overweight and obesity has been extensively reported in Western populations, little is known on the association between body weight, weight change and mortality in Asian populations whose weight distribution and mortality differ considerably from the West. METHODS: A cohort of 75 868 subjects aged 35 years and above, participants of the Trivandrum Oral Cancer Study-a cluster-randomized controlled trial originally implemented to evaluate the efficacy of visual inspection on oral cancer, in Kerala State, South India-were followed up from 1995 to 2004. Weight and height were measured both at baseline and in 3.5-year follow-up surveys. Early years of follow-up were excluded from the analyses. Relative risks of overall death and cause-specific death were estimated according to the body mass index (BMI) category of the WHO Asian population definitions, and to weight changes between two surveys. RESULTS: Low BMI was a predictor of mortality, while high BMI was not. Mortality risks in men adjusted for age, smoking habits and other potential confounders, as compared with a BMI 18.5-22.9 kg/m(2), were 1.26 (95% CI 1.03-1.55) for BMI < 16 kg/m(2); 1.16 (1.03-1.32) for BMI = 16-18.4 kg/m(2); 0.95 (0.81-1.12) for BMI = 23-24.9 kg/m(2); 0.85 (0.69-1.05) for BMI = 25-27.4 kg/m(2); and 0.89 (0.65-1.21) for BMI >/= 27.5 kg/m(2). Similar findings were observed in women. BMI was not associated with deaths from cancer, cardiovascular and cerebrovascular diseases, and diabetes. A low BMI (<16 kg/m(2)) was associated with increased deaths from chronic respiratory diseases. Smoking and socio-economical status did modify the association. A moderate weight gain of 4-10% between the two surveys was associated with decreased risk of death, while moderate and severe weight loss were predictive factors of death. Similar results were observed in both men and women. CONCLUSIONS: Among this Indian rural population, mild to severe leanness (BMI < 16 kg/m(2)) and weight loss were important determinants of mortality, especially from chronic respiratory diseases, while overweight and above (BMI > 23 kg/m(2)) did not show any detrimental effect. PMID- 18388154 TI - An integrated care pathway improves quality of life in Primary Biliary Cirrhosis. AB - BACKGROUND: Clinical management of the chronic autoimmune liver disease, Primary Biliary Cirrhosis (PBC) involves addressing the underlying liver disease and a range of symptoms independent of liver disease severity. We have formally explored how these two perspectives of chronic disease management can be combined into a clinic consultation and impact upon quality of life (QOL) in PBC. AIMS: To develop and implement the first Integrated Care Pathway (ICP) for the management of liver disease progression and symptom management in PBC. METHODS: Process mapping of current practice by a multidisciplinary group developed a flowchart of care from which the clinical record evolved. Symptom assessment is incorporated into the PBC ICP (QOL; PBC-40, autonomic symptoms; Orthostatic Grading Scale, daytime sleepiness; Epworth Sleepiness Scale). All patients were considered who attended clinic between July 2005 and June 2006. Symptom assessment was repeated after 1 year in those participating in the initial clinic cohort. RESULTS: The PBC ICP was successfully introduced into our clinical environment with high levels of patient satisfaction. A total of 225 PBC patients attended over 12 months. Initial QOL assessments were in 195 (87%). Five patients died (3%). Repeat assessment 1 year later occurred in 149 subjects (149/190; 78%). All symptom domains improved after ICP implementation with significant improvements in those with moderate and severe symptoms in all PBC-40 symptom domains (P < 0.02). In those with severe fatigue (n = 38) symptom improvement was even more dramatic (P = 0.002). CONCLUSION: ICP implementation delivers evidence-based care, leads to improvements in QOL coupled with high levels of patient satisfaction. PMID- 18388153 TI - Bone mineral density of the tarsals and metatarsals with reloading. AB - BACKGROUND AND PURPOSE: Bone mineral density (BMD) decreases rapidly with prolonged non-weight bearing. Maximizing the BMD response to reloading activities after NWB is critical to minimizing fracture risk. METHODS: for measuring individual tarsal and metatarsal BMD have not been available. This case report describes tarsal and metatarsal BMD with a reloading program, as revealed by quantitative computed tomography (QCT). CASE DESCRIPTION: A 24-year-old woman was non-weight bearing for 6 weeks after right talocrural arthroscopy. Tarsal and metatarsal BMD were measured with QCT 9 weeks (before reloading) and 32 weeks (after reloading) after surgery. A 26-week progressive reloading program was completed. Change scores were calculated for BMD before reloading and BMD after reloading for the total foot (average of all tarsals and metatarsals), tarsals, metatarsals, bones of the medial column (calcaneus, navicular, cuneiforms 1 and 2, and metatarsal 1), and bones of the lateral column (calcaneus, cuboid, cuneiform 3, and metatarsals 2-5). The percent differences in BMD between the involved side and the uninvolved side were calculated. OUTCOMES: Before reloading, BMD of the involved total foot was 9% lower than that on the uninvolved side. After reloading, BMD increased 22% and 21% for the total foot, 16% and 14% for the tarsals, 29% and 30% for the metatarsals, 14% and 15% for the medial column bones, and 28% and 26% for the lateral column bones on the involved and uninvolved sides, respectively. After reloading, BMD of the involved total foot remained 8% lower than that on the uninvolved side. DISCUSSION: The increase in BMD with reloading was not uniform across all pedal bones; the metatarsals showed a greater increase than the tarsals, and the lateral column bones showed a greater increase than the medial column bones. PMID- 18388155 TI - Right renal artery stenosis complicating Takayasu disease. PMID- 18388156 TI - Rituximab inhibits structural joint damage in patients with rheumatoid arthritis with an inadequate response to tumour necrosis factor inhibitor therapies. AB - OBJECTIVE: To determine if treatment with a B cell-targeted therapy can inhibit the progression of structural joint damage in patients with rheumatoid arthritis (RA), exhibiting an inadequate response to tumour necrosis factor (TNF) inhibitors. METHODS: In this phase III study, patients with an inadequate response to a TNF inhibitor and receiving methotrexate were randomised to rituximab or placebo. Radiographs were obtained at baseline, week 24 and week 56 after randomisation. Patients with an inadequate response to their randomised therapy could receive rescue medication from week 16. From week 24, eligible patients from both treatment arms could receive open-label rituximab. Patients were analysed according to their original treatment group. Radiographs were scored using the Genant-modified Sharp method. The primary radiographic endpoint was change in total Genant-modified Sharp score at week 56. RESULTS: Rituximab treatment caused significant reduction in joint damage progression compared with placebo. The mean change from baseline in the total Genant-modified Sharp score at week 56 was significantly lower for patients treated with rituximab than for patients treated with placebo (1.00 vs 2.31; p = 0.005), and was supported by changes in erosion score (0.59 and 1.32 for rituximab plus methotrexate vs placebo plus methotrexate, respectively; p = 0.011) and joint space narrowing score (0.41 and 0.99, respectively; p<0.001). CONCLUSIONS: This study provides the first evidence that a B cell-targeted therapy-rituximab-can significantly inhibit the progression of structural joint damage in patients with RA with long standing, active and treatment-resistant disease. PMID- 18388157 TI - The role of the shared epitope in arthralgia with anti-cyclic citrullinated peptide antibodies (anti-CCP), and its effect on anti-CCP levels. AB - OBJECTIVES: Patients presenting with both arthralgia and antibodies to cyclic citrullinated peptide (anti-CCP) have an increased risk of developing rheumatoid arthritis (RA). To further characterise this patient group and shed more light on its relationship with clinically manifest early arthritis and established RA, an immunogenetic and serological analysis was performed. METHODS: In a group of 111 patients with anti-CCP-positive arthralgia, anti-CCP levels and shared epitope (SE) status were determined. Data were compared with 125 and 128 patients with anti-CCP-positive early arthritis and established RA respectively. RESULTS: In patients with anti-CCP-positive arthralgia, the frequency of SE allele positivity is significantly lower when compared with anti-CCP-positive early arthritis and established RA (58% vs 80%, and 58% vs 92%, respectively, both p<0.001). Median anti-CCP levels were higher in the group of patients with SE-positive arthralgia compared with the group of patients with SE-negative arthralgia (p = 0.02). Median anti-CCP levels were similar in the groups of patients with SE-positive arthralgia and arthritis. CONCLUSIONS: The lower frequency of SE positivity in patients with arthralgia compared with patients with RA indicates that, compared with patients who were SE positive, patients who were SE negative as a group go through a longer arthralgia phase, or alternatively have a lower risk for transition from anti-CCP positive arthralgia to RA. Furthermore, the present results suggest that in this early stage the effect of the SE on disease risk may be mediated through higher anti-CCP levels. PMID- 18388158 TI - EULAR points to consider for conducting clinical trials in systemic lupus erythematosus. AB - OBJECTIVE: Systemic lupus erythematosus (SLE) is a complex multi-organ disease, characterised by relapses and remissions. DESIGN: ng a high-quality randomised controlled trial poses many challenges. We have developed evidenced-based recommendations for points to consider in conducting clinical trials in patients with SLE. METHODS: The EULAR Task Force on SLE comprised 19 specialists and a clinical epidemiologist. Initially, the evidence for clinical trial end-points in SLE was evaluated and this has been reported separately. A consensus approach was developed by the SLE Task Force in formulating recommendations for points to consider when conducting clinical trials in SLE. RESULTS: The literature review revealed that most outcome measures used in phase 2/3 trials in SLE have not actually been validated in clinical trials, although other forms of validation have been undertaken. The final recommendations for points to consider for conducting clinical trials in SLE address the following areas: study design, eligibility criteria, outcome measures including adverse events, concomitant therapies for SLE and its complications. CONCLUSIONS: Recommendations for points to consider when conducting clinical trials in SLE were developed using an evidence-based approach followed by expert consensus. The recommendations should be disseminated, implemented and then reviewed in detail and revised using an evidence-based approach in about 5 years, by which time there will be further evidence to consider from current clinical trials. PMID- 18388159 TI - Abnormal expression of the genes involved in cytokine networks and mitochondrial function in systemic juvenile idiopathic arthritis identified by DNA microarray analysis. AB - OBJECTIVES: Systemic juvenile idiopathic arthritis (sJIA) is a rheumatic disease in childhood characterised by systemic symptoms and a relatively poor prognosis. Peripheral leukocytes are thought to play a pathological role in sJIA although the exact cause of the disease is still obscure. In this study, we aimed to clarify cellular functional abnormalities in sJIA. METHODS: We analysed the gene expression profile in peripheral leukocytes from 51 patients with sJIA, 6 patients with polyarticular type JIA (polyJIA) and 8 healthy children utilising DNA microarrays. Gene ontology analysis and network analysis were performed on the genes differentially expressed in sJIA to clarify the cellular functional abnormalities. RESULT: A total of 3491 genes were differentially expressed in patients with sJIA compared to healthy individuals. They were functionally categorised mainly into a defence response group and a metabolism group according to gene ontology, suggesting the possible abnormalities in these functions. In the defence response group, molecules predominantly constituting interferon (IFN)gamma and tumour necrosis factor (TNF) network cascades were upregulated. In the metabolism group, oxidative phosphorylation-related genes were downregulated, suggesting a mitochondrial disorder. Expression of mitochondrial DNA-encoded genes including cytochrome c oxidase subunit 1(MT-CO1) and MT-CO2 were suppressed in patients with sJIA but not in patients with polyJIA or healthy children. However, nuclear DNA-encoded cytochrome c oxidases were intact. CONCLUSION: Our findings suggest that sJIA is not only an immunological disease but also a metabolic disease involving mitochondria disorder. PMID- 18388162 TI - Chocolate-induced prolonged angiooedema in an elderly patient. AB - This report presents a 93-year-old woman with prolonged angioedema associated with diabetic chocolate and chronic angiotensin-converting enzyme inhibitor use. PMID- 18388161 TI - Acute respiratory failure in the elderly: diagnosis and prognosis. AB - Acute respiratory failure (ARF) in patients over 65 years is common in emergency departments (EDs) and is one of the key symptoms of congestive heart failure (CHF) and respiratory disorders. Searches were conducted in MEDLINE for published studies in the English language between January 1980 and August 2007, using 'acute dyspnea', 'acute respiratory failure (ARF)', 'heart failure', 'pneumonia', 'pulmonary embolism (PE)' keywords and selecting articles concerning patients aged 65 or over. The age-related structural changes of the respiratory system, their consequences in clinical assessment and the pathophysiology of ARF are reviewed. CHF is the most common cause of ARF in the elderly. Inappropriate diagnosis that is frequent and inappropriate treatments in ED are associated with adverse outcomes. B-type natriuretic peptides (BNPs) help to determine an accurate diagnosis of CHF. We should consider non-invasive ventilation (NIV) in elderly patients hospitalised with CHF or acidotic chronic obstructive pulmonary disease (COPD) who do not improve with medical treatment. Further studies on ARF in elderly patients are warranted. PMID- 18388160 TI - MRI bone oedema is the strongest predictor of subsequent radiographic progression in early rheumatoid arthritis. Results from a 2-year randomised controlled trial (CIMESTRA). AB - OBJECTIVE: To identify predictors of radiographic progression in a 2-year randomised, double-blind, clinical study (CIMESTRA) of patients with early rheumatoid arthritis (RA). METHODS: Patients with early RA (n = 130) were treated with methotrexate, intra-articular betamethasone and ciclosporin/placebo ciclosporin. Baseline magnetic resonance imaging (MRI) of the wrist (wrist-only group, n = 130) or MRI of wrist and metacarpophalangeal (MCP) joints (wrist+MCP group, n = 89) (OMERACT RAMRIS), x-ray examination of hands, wrists and forefeet (Sharp/van der Heijde Score (TSS)), Disease Activity Score (DAS28), anti-cyclic citrullinated peptide antibodies (anti-CCP), HLA-DRB1-shared epitope (SE) and smoking status were assessed. Multiple regression analysis was performed with delta-TSS (0-2 years) as dependent variable and baseline DAS28, TSS, MRI bone oedema score, MRI synovitis score, MRI erosion score, anti-CCP, smoking, SE, age and gender as explanatory variables. RESULTS: Baseline values: median DAS28 5.6 (range 2.4-8.0); anti-CCP positive 61%; radiographic erosions 56%. At 2 years: DAS28 2.0 (0.5-5.7), in DAS remission: 56%, radiographic progression 26% (wrist+MCP group, similar for wrist-only group). MRI bone oedema score was the only independent predictor of delta-TSS (wrist+MCP group: coefficient = 0.75 (95% CI 0.55 to 0.94), p<0.001; wrist-only group: coefficient = 0.59 (95% CI 0.40 to 0.77), p<0.001). Bone oedema score explained 41% of the variation in the progression of TSS (wrist+MCP group), 25% in wrist-only group (Pearson's r = 0.64 and r = 0.50, respectively). Results were confirmed by sensitivity analyses. CONCLUSION: In a randomised controlled trial aiming at remission in patients with early RA, baseline RAMRIS MRI bone oedema score of MCP and wrist joints (and of wrist only) was the strongest independent predictor of radiographic progression in hands, wrists and forefeet after 2 years. MRI synovitis score, MRI erosion score, DAS28, anti-CCP, SE, smoking, age and gender were not independent risk factors. TRIAL REGISTRATION NUMBER: NCT00209859. PMID- 18388163 TI - Nursing ethics. PMID- 18388164 TI - The wounded healer - what has the concept to offer nursing? PMID- 18388165 TI - A concept development of 'being sensitive' in nursing. AB - 'Being sensitive' in nursing was explored using Schwartz-Barcott and Kim's hybrid model of concept development, producing a tentative definition of the concept. Three phases were employed: theoretical, empirical/fieldwork and analytical. An exploration of the literature identified where the common idea of ;being sensitive' as a nurse was embedded and demonstrated that a theoretical development of this fundamental aspect of nursing was absent. The empirical phase was conducted using semistructured interviews with nine expert palliative care and cancer nurses. This method was particularly useful for the exploration of this concept because of its firm grounding in practical example. A definition of what the concept ;being sensitive' means in nursing, and subsequent clarification of ;being insensitive', have been posed from the research process undertaken. The essential nature of this concept being integral to nursing practice is emphasized. Potential implications for the development of nursing practice through teaching of this concept were identified. PMID- 18388166 TI - Nurses' moral sensitivity and hospital ethical climate: a literature review. AB - Increased technological and pharmacological interventions in patient care when patient outcomes are uncertain have been linked to the escalation in moral and ethical dilemmas experienced by health care providers in acute care settings. Health care research has shown that facilities that are able to attract and retain nursing staff in a competitive environment and provide high quality care have the capacity for nurses to process and resolve moral and ethical dilemmas. This article reports on the findings of a systematic review of the empirical literature (1980 - February 2007) on the effects of unresolved moral distress and poor ethical climate on nurse turnover. Articles were sought to answer the review question: Does unresolved moral distress and a poor organizational ethical climate increase nurse turnover? Nine articles met the criteria of the review process. Although the prevailing sentiment was that poor ethical climate and moral distress caused staff turnover, definitive answers to the review question remain elusive because there are limited data that confidently support this statement. PMID- 18388167 TI - Learning, decisions and transformation in critical care nursing practice. AB - Critical care nurses are key providers in a high acuity environment. This qualitative research study explored ethical decision making in a critical care practice setting. Fifteen critical care nurses with varying experience and education levels were purposively sampled to assure the representativeness of the data. The theoretical concepts of experiential learning, perspective transformation, reflection-in-action and principle-based ethics were used as a framework for eliciting information from the participants. A new model of focused reflection in ethical decision making was developed. Findings showed that having a role model or mentor to help guide the ethical decision-making process was critical for focused ethical discourse and the decision making. PMID- 18388168 TI - Nursing leaders' experiences with the ethical dimensions of nursing education. AB - This pilot study explores four nursing leaders' experiences with the ethical dimensions of leadership in education. Gathering and interpreting such data of experience fosters greater understanding of the nature of moral leadership as it is lived in nursing education. A phenomenological approach was used to collect and analyze the data. The results revealed four major themes: integrity, justice, wrestling with decisions in the light of consequences, and the power of information. These themes clarify the values that direct these leaders' actions as they mediate community needs common to educational and health care institutions. PMID- 18388169 TI - Nursing students' perceptions of self-determination in elderly people. AB - The purpose of this study was to compare nursing students' perceptions of self determination in elderly patients before and after clinical training in long term care institutions as a part of their course in gerontological nursing. A pre- post-test design was employed. The data were collected by questionnaires completed by students at one nurse education organization college in Finland (pre test n +/- 120, response rate 95%; post-test n +/- 115, response rate 91%). The students' perceptions of elderly patients' self-determination were more positive after their clinical training period concerning to what extent elderly patients are able to control their treatment and what kind of support they received from nurses to exercise their self-determination. The students' perceptions remained stable concerning how important self-determination is to elderly patients, and how willing and knowledgeable they are about using their self-determination. Ethics teaching, together with high quality clinical training placements, should be assured early during nursing studies. PMID- 18388170 TI - Autonomy and advocacy in perinatal nursing practice. AB - Advocacy has been positioned as an ideal within the practice of nursing, with national guidelines and professional standards obliging nurses to respect patients' autonomous choices and to act as their advocates. However, the meaning of advocacy and autonomy is not well defined or understood, leading to uncertainty regarding what is required, expected and feasible for nurses in clinical practice. In this article, a feminist ethics perspective is used to examine how moral responsibilities are enacted in the perinatal nurse-patient relationship and to explore the interaction between the various threads that influence, and are in turn affected by, this relationship. This perspective allows for consideration of contextual and relational factors that impact on the way perinatal nursing care is given and received, and provides a framework for exploring the ways in which patient autonomy, advocacy and choice are experienced by childbearing women and their nurses during labour and birth. PMID- 18388171 TI - Nurses as guests or professionals in home health care. AB - The aim of this study was to explore and interpret the diverse subject of positions, or roles, that nurses construct when caring for patients in their own home. Ten interviews were analysed and interpreted using discourse analysis. The findings show that these nurses working in home care constructed two positions: ;guest' and ;professional'. They had to make a choice between these positions because it was impossible to be both at the same time. An ethics of care and an ethics of justice were present in these positions, both of which create diverse ethical appeals, that is, implicit demands to perform according to a guest or to a professional norm. PMID- 18388172 TI - Achieving care and social justice for people with dementia. AB - This article draws on two studies that have used an ethic of care analysis to explore lay, nursing and social work care for people with dementia. It discusses the political as well as the practice application of ethic of care principles and highlights the necessity to understand both what people do and the meanings with which such practices are imbued in order to identify ;good care' and the relationship between this and social justice. Examples of care for people with dementia are discussed by reference to core principles of an ethic of care: attentiveness, responsibility, competence, responsiveness and trust. These illustrate the potential for the development of a shared language within which different disciplines, lay carers and people with dementia can communicate about how needs could best be met in complex and difficult circumstances. PMID- 18388173 TI - Exposing nursing students to the marketing methods of pharmaceutical companies. AB - There is a strong association between reliance on the promotional activities of pharmaceutical companies and a generally less appropriate use of prescription drugs. Pharmaceutical companies direct some of their promotion towards health workers who do not have the authority to prescribe medicines, such as nurses in certain countries. The aim of this study was to determine the impact that exposure to the marketing methods of pharmaceutical companies has on judgments made by nursing students about health worker-pharmaceutical company relationships. A cross-sectional survey was carried out with 442 nursing students in Istanbul, Turkey. The exposure of students to the marketing methods of pharmaceutical companies, whether it be indirectly through observation or directly by first-hand experience, increases the probability that students will adopt rationales that underlie affirmative judgments of health worker pharmaceutical company relationships. Based on the pervasiveness and ability of drug promotion to influence the perceptions of students, it is imperative that attempts be made to reduce its negative impact. PMID- 18388174 TI - Virginia Tilden: interviewed by Anne Davis. PMID- 18388175 TI - Dosemeter readings and effective dose to the cardiologist with protective clothing in a simulated interventional procedure. AB - A personal dosemeter issued for individual monitoring is calibrated in terms of personal dose equivalent, usually H(P)(10). In general it yields a reasonable estimate of effective dose (E) when the exposed person does not wear protective clothing. In interventional cardiology, however, a lead equivalent apron is worn and often a thyroid collar. A correction factor will then be necessary to convert a dosemeter reading to E. To explore this factor an interventional cardiology procedure is simulated based on exposure conditions typical for a modern hospital in the BENELUX area. The dose to the cardiologist is investigated using Monte Carlo simulation of radiation transport. It is concluded that a personal dosemeter may best be worn outside the apron at a central position high on the chest for least dependence on the beam direction. It will overestimate E by roughly a factor of 20 (apron and thyroid collar of 0.25 mm Pb). PMID- 18388176 TI - Narcolepsy in Southern Chinese patients: clinical characteristics, HLA typing and seasonality of birth. AB - OBJECTIVE: To report clinical characteristics, human leukocyte antigen (HLA) typing and seasonality of birth of a series of 54 Southern Chinese patients suffering from narcolepsy. METHODS: All subjects underwent detailed medical and psychiatric interviews and a standardised nocturnal polysomnogram followed by a daytime Multiple Sleep Latency Test. Each subject also completed a set of sleep questionnaires. HLA typing was performed in 91% of subjects. RESULTS: A total of 78% and 22% of patients were diagnosed with suffering from cataplectic and non cataplectic narcolepsy, respectively. The majority (n = 47, 87%) of patients were referred to our sleep clinic for excessive daytime sleepiness (EDS). The cataplectic narcolepsy differed from non-cataplectic narcolepsy by having more rapid eye movement (REM)-related clinical symptoms (more sleep paralysis and sleep-related hallucination) and sleep disturbances (shorter REM latency), as well as tighter association with HLA DQB1*0602. A bi-modal peak pattern was observed at 11 and 39 years old. A similar bi-modal pattern also occurred for EDS and cataplexy. Excess winter births were observed for this series of patients. 81% of patients with cataplectic narcolepsy were DQB1*0602-positive. There were no differences between early- and late-onset cases in the association with positive DQB1*0602 (71.4% vs 60%). Narcolepsy had prominent pernicious effects on various social, academic, family and mental aspects in our patients. CONCLUSIONS: In our Southern Chinese narcolepsy series, bi-modal peak pattern of age of onset, excess winter birth and tight association of HLA DQB1*0602 with cataplectic narcolepsy were found. PMID- 18388177 TI - Effect of plasma protein binding on in vitro-in vivo correlation of biliary excretion of drugs evaluated by sandwich-cultured rat hepatocytes. AB - In the present study, we examined in vitro biliary clearance of several compounds in sandwich-cultured rat hepatocytes (SCRH) and compared it with that observed in vivo in rats; the effect of plasma protein binding on in vitro-in vivo correlation of biliary excretion was also assessed. The in vitro biliary excretion was determined by differential cumulative uptake of compounds in SCRH preincubated in the presence and absence of Ca(2+)/Mg(2+). The cumulative uptake study of radiolabeled substrates revealed that the function of canalicular efflux transporters such as bile salt export pump, multidrug resistance-associated protein 2, breast cancer resistance protein, and multidrug resistance 1 was adequately maintained in SCRH. Unlabeled test compounds, pravastatin, rosuvastatin, valsartan, cefmetazole, and cefoperazone exhibited varying degrees of in vitro biliary excretion in the cumulative uptake study using SCRH. In vivo biliary excretions of these compounds were measured in common bile duct cannulated rats. Whereas their biliary excretion ratios were all more than 60% of the dose, the in vivo intrinsic biliary clearances varied from 10.5 to 1787.2 ml/min/kg. The in vitro intrinsic biliary clearances of test compounds were well correlated with their corresponding in vivo intrinsic clearances calculated on the basis of the plasma unbound concentration (r(2) = 0.984), whereas less correlation was observed when they were calculated on the basis of plasma total concentration (r(2) = 0.217). These results indicate that SCRH is a useful in vitro model for predicting in vivo intrinsic biliary clearance in rats. In addition, for an accurate prediction, it is necessary to evaluate the in vivo intrinsic biliary clearance based on plasma unbound concentration but not total concentration. PMID- 18388179 TI - Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors. AB - Human embryonic stem cells (hESCs) could potentially represent an alternative source for blood transfusion therapies and a promising tool for studying the ontogeny of hematopoiesis. When we cultured hESCs on either C3H10T1/2 or OP-9 cells to facilitate hematopoiesis, we found that exogenous administration of vascular endothelial growth factor promoted the emergence of sac-like structures, which we named embryonic stem cell-derived sacs (ES-sacs). These ES-sacs consisted of multiple cysts demarcated by cellular monolayers that retained some of the properties of endothelial cells. The spherical cells inside ES-sacs expressed primarily CD34, along with VE-cadherin, CD31, CD41a, and CD45, and were able to form hematopoietic colonies in semisolid culture and to differentiate into mature megakaryocytes by day 24 in the presence of thrombopoietin. Apparently, ES-sacs provide a suitable environment for hematopoietic progenitors. Relatively large numbers of mature megakaryocytes could be induced from the hematopoietic progenitors within ES-sacs, which were then able to release platelets that displayed integrin alpha IIb beta 3 activation and spreading in response to ADP or thrombin. This novel protocol thus provides a means of generating platelets from hESCs, which could serve as the basis for efficient production of platelets for clinical transfusion and studies of thrombopoiesis. PMID- 18388178 TI - Clinical significance of minimal residual disease in childhood acute lymphoblastic leukemia and its relationship to other prognostic factors: a Children's Oncology Group study. AB - Minimal residual disease (MRD) is an important predictor of relapse in acute lymphoblastic leukemia (ALL), but its relationship to other prognostic variables has not been fully assessed. The Children's Oncology Group studied the prognostic impact of MRD measured by flow cytometry in the peripheral blood at day 8, and in end-induction (day 29) and end-consolidation marrows in 2143 children with precursor B-cell ALL (B-ALL). The presence of MRD in day-8 blood and day-29 marrow MRD was associated with shorter event-free survival (EFS) in all risk groups; even patients with 0.01% to 0.1% day-29 MRD had poor outcome compared with patients negative for MRD patients (59% +/- 5% vs 88% +/- 1% 5-year EFS). Presence of good prognostic markers TEL-AML1 or trisomies of chromosomes 4 and 10 still provided additional prognostic information, but not in National Cancer Institute high-risk (NCI HR) patients who were MRD(+). The few patients with detectable MRD at end of consolidation fared especially poorly, with only a 43% plus or minus 7% 5-year EFS. Day-29 marrow MRD was the most important prognostic variable in multi-variate analysis. The 12% of patients with all favorable risk factors, including NCI risk group, genetics, and absence of days 8 and 29 MRD, had a 97% plus or minus 1% 5-year EFS with nonintensive therapy. These studies are registered at www.clinicaltrials.gov as NCT00005585, NCT00005596, and NCT00005603. PMID- 18388180 TI - Efficient transduction of pigtailed macaque hematopoietic repopulating cells with HIV-based lentiviral vectors. AB - Lentiviral vectors are attractive for hematopoietic stem cell (HSC) gene therapy because they do not require mitosis for nuclear entry, they efficiently transduce hematopoietic repopulating cells, and self-inactivating (SIN) designs can be produced at high titer. Experiments to evaluate HIV-derived lentiviral vectors in nonhuman primates prior to clinical trials have been hampered by low transduction frequencies due in part to host restriction by TRIM5alpha. We have established conditions for efficient transduction of pigtailed macaque (Macaca nemestrina) long-term repopulating cells using VSV-G-pseudotyped HIV-based lentiviral vectors. Stable, long-term, high-level gene marking was observed in 3 macaques using relatively low MOIs (5-10) in a 48-hour ex vivo transduction protocol. All animals studied had rapid neutrophil engraftment with a median of 10.3 days to a count greater than 0.5 x 10(9)/L (500/microL). Expression was detected in all lineages, with long-term marking levels in granulocytes at approximately 20% to 30%, and in lymphocytes at approximately 12% to 23%. All animals had polyclonal engraftment as determined by analysis of vector integration sites. These data suggest that lentiviral vectors should be highly effective for HSC gene therapy, particularly for diseases in which maintaining the engraftment potential of stem cells using short-term ex vivo transduction protocols is critical. PMID- 18388181 TI - Leukemogenic mechanisms and targets of a NUP98/HHEX fusion in acute myeloid leukemia. AB - We have studied a patient with acute myeloid leukemia (AML) and t(10;11)(q23;p15) as the sole cytogenetic abnormality. Molecular analysis revealed a translocation involving nucleoporin 98 (NUP98) fused to the DNA-binding domain of the hematopoietically expressed homeobox gene (HHEX). Expression of NUP98/HHEX in murine bone marrow cells leads to aberrant self-renewal and a block in normal differentiation that depends on the integrity of the NUP98 GFLG repeats and the HHEX homeodomain. Transplantation of bone marrow cells expressing NUP98/HHEX leads to transplantable acute leukemia characterized by extensive infiltration of leukemic blasts expressing myeloid markers (Gr1(+)) as well as markers of the B cell lineage (B220(+)). A latency period of 9 months and its clonal character suggest that NUP98/HHEX is necessary but not sufficient for disease induction. Expression of EGFP-NUP98/HHEX fusions showed a highly similar nuclear localization pattern as for other NUP98/homeodomain fusions, such as NUP98/HOXA9. Comparative gene expression profiling in primary bone marrow cells provided evidence for the presence of common targets in cells expressing NUP98/HOXA9 or NUP98/HHEX. Some of these genes (Hoxa5, Hoxa9, Flt3) are deregulated in NUP98/HHEX-induced murine leukemia as well as in human blasts carrying this fusion and might represent bona fide therapeutic targets. PMID- 18388182 TI - CD4+ CD25+ Treg cells inhibit human memory gammadelta T cells to produce IFN gamma in response to M tuberculosis antigen ESAT-6. AB - Gammadelta T cells play an important role in innate immunity against infections; however, the regulation of these cells remains largely unknown. In the present study, we show that ESAT-6, an antigen of Mycobacterium tuberculosis, induces IFN gamma secretion by human gammadelta T cells. In addition, ESAT-6 also induces the activation and proliferation of gammadelta T cells. Phenotypic analysis indicates that IFN-gamma-producing gammadelta T cells are mainly effector memory cells with the surface phenotype of CD45RA(-)CD62L(-)CCR7(-). These results were further confirmed by the fact that naive gammadelta T cells from cord blood did not produce IFN-gamma in response to ESAT-6. Further studies indicated that stimulation with ESAT-6 directly induced purified gammadelta T cells to produce IFN-gamma, independent of both antigen-presenting cells and CD4(+) T cells. Unexpectedly, depletion of CD4(+) T cells markedly enhanced IFN-gamma production by gammadelta T cells, indicating that CD4(+) T cells regulate the response of gammadelta T cells. Importantly, CD4(+)CD25(+) T regulatory (Treg) cells but not CD4(+)CD25(-) T cells significantly inhibited IFN-gamma production by gammadelta T cells. Taken together, these data demonstrate for the first time that Treg cells can play an important role in the regulation of immune responses of antigen specific human memory gammadelta T cells. PMID- 18388183 TI - The novel protein kinase C isoforms -delta and -epsilon modulate caerulein induced zymogen activation in pancreatic acinar cells. AB - Isoforms of protein kinase C (PKC) have been shown to modulate some cellular responses such as pathological secretion and generation of inflammatory mediators during acute pancreatitis (AP). We propose that PKC also participates in premature zymogen activation within the pancreatic acinar cell, a key event in the initiation of AP. This hypothesis was examined in in vivo and cellular models of caerulein-induced AP using PKC activators and inhibitors. Phorbol ester, 12-O tetradecanoylphorbol-13-acetate (TPA, 200 nM), a known activator of PKC, enhanced zymogen activation at both 0.1 nM and 100 nM caerulein, concentrations which mimic physiological and supraphysiological effects of the hormone cholecystokinin, respectively, in preparations of pancreatic acinar cells. Isoform-specific PKC inhibitors for PKC-delta and PKC-epsilon reduced supraphysiological caerulein-induced zymogen activation. Using a cell-free reconstitution system, we showed that inhibition of PKC-delta and -epsilon, reduced zymogen activation in both zymogen granule-enriched and microsomal fractions. In dispersed acinar cells, 100 nM caerulein stimulation caused PKC delta and -epsilon isoform translocation to microsomal membranes using cell fractionation and immunoblot analysis. PKC translocation was confirmed with in vivo studies and immunofluorescence microscopy in pancreatic tissues from rats treated with or without 100 nM caerulein. PKC-epsilon redistributed from an apical to a supranuclear region following caerulein administration. The signal for PKC-epsilon overlapped with granule membrane protein, GRAMP-92, an endosomal/lysosomal marker, in a supranuclear region where zymogen activation takes place. These results indicate that PKC-delta and -epsilon isoforms translocate to specific acinar cell compartments and modulate zymogen activation. PMID- 18388184 TI - Cellular cross talk in the small intestinal mucosa: postnatal lymphocytic immigration elicits a specific epithelial transcriptional response. AB - During the early postnatal period lymphocytes migrate into the mouse small intestine. Migrating infiltrative lymphocytes have the potential to affect the epithelial cells via secreted cytokines. Such cross talk can result in the elicitation of an epithelial transcriptional response. Knowledge about such physiological cross talk between the immune system and the epithelium in the postnatal small intestinal mucosa is lacking. We have investigated the transcriptome changes occurring in the postnatal mouse small intestine using DNA microarray technology, immunocytochemistry, and quantitative real-time RT-PCR analysis. The DNA microarray data were analyzed bioinformatically by using a combination of projections to latent structures analysis and functional annotation analysis. The results show that infiltrating lymphocytes appear in the mouse small intestine in the late postweaning period and give rise to distinct changes in the epithelial transcriptome. Of particular interest is the expression of three genes encoding a mucin (Muc4), a mucinlike protein (16000D21Rik), and ATP citrate lyase (Acly). All three genes were shown to be expressed by the epithelium and to be upregulated in response to lymphocytic migration into the small intestinal mucosa. PMID- 18388185 TI - Gene expression in human NAFLD. AB - Despite the high prevalence of nonalcoholic fatty liver disease (NAFLD), little is known of its pathogenesis based on study of human liver samples. By the use of Affymetrix GeneChips (17,601 genes), we investigated gene expression in the human liver of subjects with extreme steatosis due to NAFLD without histological signs of inflammation (liver fat 66.0 +/- 6.8%) and in subjects with low liver fat content (6.4 +/- 2.7%). The data were analyzed by using sequence-based reannotation of Affymetrix probes and a robust model-based normalization method. We identified genes involved in hepatic glucose and lipid metabolism, insulin signaling, inflammation, coagulation, and cell adhesion to be significantly associated with liver fat content. In addition, genes involved in ceramide signaling (MAP2K4) and metabolism (UGCG) were found to be positively associated with liver fat content. Genes involved in lipid metabolism (PLIN, ACADM), fatty acid transport (FABP4, CD36), amino acid catabolism (BCAT1), and inflammation (CCL2) were validated by real-time PCR and were found to be upregulated in subjects with high liver fat content. The data show that multiple changes in gene expression characterize simple steatosis. PMID- 18388186 TI - PI3K/Akt activation is critical for early hepatic regeneration after partial hepatectomy. AB - Hepatic resection is associated with rapid proliferation and regeneration of the remnant liver. Phosphatidylinositol 3-kinase (PI3K), composed of a p85alpha regulatory and a p110alpha catalytic subunit, participates in multiple cellular processes, including cell growth and survival; however, the role of PI3K in liver regeneration has not been clearly delineated. In this study, we used the potent PI3K inhibitor wortmannin and small interfering RNA (siRNA) targeting the p85alpha and p110alpha subunits to determine whether total or selective PI3K inhibition would abrogate the proliferative response of the liver after partial hepatectomy in mice. Hepatic resection is associated with an induction in PI3K activity; total PI3K blockade with wortmannin and selective inhibition of p85alpha or p110alpha with siRNA resulted in a significant decrease in hepatocyte proliferation, especially at the earliest time points. Fewer macrophages and Kupffer cells were present in the regenerating liver of mice treated with wortmannin or siRNA to p85alpha or p110alpha, as reflected by a paucity of F4/80 positive cells. Additionally, PI3K inhibition led to an aberrant architecture in the regenerating hepatocytes characterized by vacuolization, lipid deposition, and glycogen accumulation; these changes were not noted in the sham livers. Our data demonstrate that PI3K/Akt pathway activation plays a critical role in the early regenerative response of the liver after resection; inhibition of this pathway markedly abrogates the normal hepatic regenerative response, most likely by inhibiting macrophage infiltration and cytokine elaboration and thus hepatocyte priming for replication. PMID- 18388187 TI - Functional characterization, localization, and molecular identification of rabbit intestinal N-amino acid transporter. AB - We have characterized the Na-glutamine cotransporter in the rabbit intestinal crypt cell brush border membrane vesicles (BBMV). Substrate specificity experiments showed that crypt cell glutamine uptake is mediated by system N. Real time PCR experiments showed that SN2 (SLC38A5) mRNA is more abundant in crypt cells compared with SN1 (SLC38A3), indicating that SN2 is the major glutamine transporter present in the apical membrane of the crypt cells. SN2 cDNA was obtained by screening a rabbit intestinal cDNA library with human SN1 used as probe. Rabbit SN2 cDNA encompassed a 473-amino-acid-long open reading frame. SN2 protein displayed 87% identity and 91% similarity to human SN2. Functional characterization studies of rabbit SN2 were performed by using vaccinia virus mediated transient expression system. Substrate specificity of the cloned transporter was identical to that of SN2 described in the literature and matched well with substrate specificity experiments performed using crypt cell BBMV. Cloned rabbit SN2, analogous to its human counterpart, is Li(+) tolerant. Hill coefficient for Li(+) activation of rabbit SN2-mediated uptake was 1. Taken together, functional data from the crypt cell BBMV and the cloned SN2 cDNA indicate that the crypt cell glutamine transport is most likely mediated by SN2. PMID- 18388188 TI - Cyclic AMP accelerates calcium waves in pancreatic acinar cells. AB - Cytosolic Ca(2+) (Ca(i)(2+)) flux within the pancreatic acinar cell is important both physiologically and pathologically. We examined the role of cAMP in shaping the apical-to-basal Ca(2+) wave generated by the Ca(2+)-activating agonist carbachol. We hypothesized that cAMP modulates intra-acinar Ca(2+) channel opening by affecting either cAMP-dependent protein kinase (PKA) or exchange protein directly activated by cAMP (Epac). Isolated pancreatic acinar cells from rats were stimulated with carbachol (1 muM) with or without vasoactive intestinal polypeptide (VIP) or 8-bromo-cAMP (8-Br-cAMP), and then Ca(i)(2+) was monitored by confocal laser-scanning microscopy. The apical-to-basal carbachol (1 muM) stimulated Ca(2+) wave was 8.63 +/- 0.68 microm/s; it increased to 19.66 +/- 2.22 microm/s (*P < 0.0005) with VIP (100 nM), and similar increases were observed with 8-Br-cAMP (100 microM). The Ca(2+) rise time after carbachol stimulation was reduced in both regions but to a greater degree in the basal. Lag time and maximal Ca(2+) elevation were not significantly affected by cAMP. The effect of cAMP on Ca(2+) waves also did not appear to depend on extracellular Ca(2+). However, the ryanodine receptor (RyR) inhibitor dantrolene (100 microM) reduced the cAMP-enhancement of wave speed. It was also reduced by the PKA inhibitor PKI (1 microM). 8-(4-chloro-phenylthio)-2'-O-Me-cAMP, a specific agonist of Epac, caused a similar increase as 8-Br-cAMP or VIP. These data suggest that cAMP accelerates the speed of the Ca(2+) wave in pancreatic acinar cells. A likely target of this modulation is the RyR, and these effects are mediated independently by PKA and Epac pathways. PMID- 18388189 TI - In adrenal glomerulosa cells, angiotensin II inhibits proliferation by interfering with fibronectin-integrin signaling. AB - Angiotensin II (Ang II), through the Ang II type 1 receptor subtype, inhibits basal proliferation of adrenal glomerulosa cells by inducing the disruption of actin stress fiber organization. This effect is observed in cells cultured on plastic or on fibronectin. The aim of the present study was to investigate how Ang II may interfere with extracellular matrix/integrin signaling. In cells treated for 3 d with echistatin (EC) (a snake-venom RGD-containing protein that abolishes fibronectin binding to alpha(5)beta(1) or alpha(v)beta(3) integrins), basal proliferation decreased by 38%, whereas Ang II was unable to abolish basal proliferation. In cells grown on fibronectin, Ang II decreased binding of paxillin to focal adhesions and, similarly to EC, induced a rapid dephosphorylation of paxillin (1 min), followed by an increase after 15 min. Fibronectin enhanced RhoA/B and Rac activation induced by Ang II, an effect abolished by EC. Under basal conditions, paxillin was more readily associated with RhoA/B than with Rac. Stimulation with Ang II induced a transient decrease in RhoA/B-associated paxillin (after 5 min), with a return to basal levels after 10 min, while increasing Rac-associated paxillin. Finally, results reveal that glomerulosa cells are able to synthesize and secrete fibronectin, a process by which cells can stimulate their own proliferative activity when cultured on plastic. Together, these results suggest that Ang II acts at the level of integrin-paxillin complexes to disrupt the well- developed microfilament network, a condition necessary for the inhibition of cell proliferation and initiation of steroidogenesis. PMID- 18388190 TI - Relaxin ameliorates fibrosis in experimental diabetic cardiomyopathy. AB - Fibrosis (extracellular matrix accumulation) is the final end point in diabetic cardiomyopathy. The current study evaluated the therapeutic effects of the antifibrotic hormone relaxin (RLX) in streptozotocin-treated transgenic mRen-2 rats, which undergo pathological and functional features similar to human diabetes. Twelve-week-old hyperglycemic mRen-2 rats, normoglycemic control rats, and animals treated with recombinant human gene-2 (H2) RLX from wk 10-12 were assessed for various measures of left ventricular (LV) fibrosis, hemodynamics, and function, while the mechanism of RLX's actions was also determined. Hyperglycemic mRen-2 rats had increased LV collagen concentration (fibrosis) and gelatinase activity (all P < 0.05 vs. controls) but equivalent levels of interstitial collagenase and tissue inhibitor of metalloproteinase-1 to that measured in control rats. The increased LV fibrosis associated with diabetic animals led to significant alterations in the E/A wave ratio and E-wave deceleration time (both P < 0.05 vs. controls) in the absence of blood pressure changes, reflective of myocardial stiffness and LV diastolic dysfunction. H2-RLX treatment of diabetic rats led to significant decreases in interstitial and total LV collagen deposition (both P < 0.05 vs. diabetic group), resulting in decreased myocardial stiffness and improved LV diastolic function, without affecting nondiabetic animals. The protective effects of H2-RLX in diabetic rats were associated with a reduction in mesenchymal cell differentiation and tissue inhibitor of metalloproteinase-1 expression in addition to a promotion of extracellular matrix-degrading matrix metalloproteinase-13 (all P < 0.05 vs. diabetic group) but were independent of blood pressure regulation. These findings demonstrate that RLX is an antifibrotic with rapid-occurring efficacy and may represent a novel therapy for the treatment of diabetes. PMID- 18388191 TI - Identification of key amino acid residues in a thyrotropin receptor monoclonal antibody epitope provides insight into its inverse agonist and antagonist properties. AB - CS-17 is a murine monoclonal antibody to the human TSH receptor (TSHR) with both inverse agonist and antagonist properties. Thus, in the absence of ligand, CS-17 reduces constitutive TSHR cAMP generation and also competes for TSH binding to the receptor. The present data indicate that for both of these functions, the monovalent CS-17 Fab (50 kDa) behaves identically to the intact, divalent IgG molecule (150 kDa). The surprising observation that CS-17 competes for TSH binding to the human but not porcine TSHR enabled identification of a number of amino acids in its epitope. Replacement of only three human TSHR residues (Y195, Q235, and S243) with the homologous porcine TSHR residues totally abolishes CS-17 binding as detected by flow cytometry. TSH binding is unaffected. Of these residues, Y195 is most important, with Q235 and S243 contributing to CS-17 binding to a much lesser degree. The functional effects of CS-17 IgG and Fab on constitutive cAMP generation by porcinized human TSHR confirm the CS-17 binding data. The location of TSHR amino acid residues Y195, Q235, and S243 deduced from the crystal structure of the FSH receptor leucine-rich domain provides valuable insight into the CS-17 and TSH binding sites. Whereas hormone ligands bind primarily to the concave surface of the leucine-rich domains, a major portion of the CS-17 epitope lies on the opposite convex surface with a minor component in close proximity to known TSH binding residues. PMID- 18388192 TI - Contributions of specificity protein-1 and steroidogenic factor 1 to Adcy4 expression in Y1 mouse adrenal cells. AB - The type 4 adenylyl cyclase, Adcy4, is the least abundant of five different adenylyl cyclase isoforms expressed in the Y1 mouse adrenocortical cell line and is deficient in a Y1 mutant with impaired steroidogenic factor 1 (SF1) activity. This study examines the contributions of SF1 and other DNA promoter/regulatory elements to Adcy4 expression in the Y1 cell line and its derivative Adcy4 deficient mutant. Primer extension and in silico analyses indicate that Adcy4 transcription initiates from multiple sites just downstream of a GC-rich sequence. Luciferase reporter gene assays identify a 124-bp sequence, situated 19 bp upstream of the major transcription start site and highly conserved among several mammalian species, as the major determinant of Adcy4 expression in Y1 cells and as a site of compromised activity in the Adcy4-deficient mutant. EMSAs using competitor nucleotides and specific antibodies indicate that this conserved region contains three specificity protein (Sp)-1/Sp3-binding sites and one SF1 binding site. As determined by site-specific mutagenesis, the 5'-most Sp1/Sp3 site enhances promoter activity, whereas the middle Sp1/Sp3 and SF1 sites each repress Adcy4 promoter activity. In the Adcy4-deficient mutant, mutating the SF1 site restores Adcy4 promoter activity and knocking down SF1 with small interfering RNAs increases Adcy4 expression, confirming the contribution of SF1 to the mutant phenotype. These studies demonstrate roles for Sp1/Sp3 and SF1 in Adcy4 expression in Y1 cells and establish a repressor function for SF1 in certain promoter contexts. PMID- 18388193 TI - Epithelial sodium channel is a key mediator of growth hormone-induced sodium retention in acromegaly. AB - Acromegalic patients present with volume expansion and arterial hypertension, but the renal sites and molecular mechanisms of direct antinatriuretic action of GH remain unclear. Here, we show that acromegalic GC rats, which are chronically exposed to very high levels of GH, exhibited a decrease of furosemide-induced natriuresis and an increase of amiloride-stimulated natriuresis compared with controls. Enhanced Na(+),K(+)-ATPase activity and altered proteolytic maturation of epithelial sodium channel (ENaC) subunits in the cortical collecting ducts (CCDs) of GC rats provided additional evidence for an increased sodium reabsorption in the late distal nephron under chronic GH excess. In vitro experiments on KC3AC1 cells, a murine CCD cell model, revealed the expression of functional GH receptors and IGF-I receptors coupled to activation of Janus kinase 2/signal transducer and activator of transcription 5, ERK, and AKT signaling pathways. That GH directly controls sodium reabsorption in CCD cells is supported by: 1) stimulation of transepithelial sodium transport inhibited by GH receptor antagonist pegvisomant; 2) induction of alpha-ENaC mRNA expression; and 3) identification of signal transducer and activator of transcription 5 binding to a response element located in the alpha-ENaC promoter, indicative of the transcriptional regulation of alpha-ENaC by GH. Our findings provide the first evidence that GH, in concert with IGF-I, stimulates ENaC-mediated sodium transport in the late distal nephron, accounting for the pathogenesis of sodium retention in acromegaly. PMID- 18388194 TI - Oxidative stress stimulates testicular orphan receptor 4 through forkhead transcription factor forkhead box O3a. AB - Early studies reveal that testicular orphan nuclear receptor 4 (TR4) modulates signaling pathways that control various cell functions. However, how TR4 activity is regulated without the involvement of specific ligand(s) remains unclear. Here we identify a daf-16 family protein-binding element (DBE; 5'-TGTTTAC-3') in the TR4 promoter that can be recognized by the forkhead transcriptional factor FOXO3a, a key stress-responsive factor, through which TR4 gene expression is activated. The interaction between DBE and FOXO3a was confirmed using EMSA and chromatin immunoprecipitation assays. Activation of FOXO3a by oxidative stress and phosphatidylinositol 3-kinase inhibitor induced TR4 expression; in contrast, suppression of FOXO3a by small interfering RNA can reduce oxidative stress induced TR4 expression. The biological consequence of the FOXO3a-induced TR4 by oxidative stress is to protect against stress-induced cell death in which cells with reduced FOXO3a are less resistant to oxidative stress, and addition of functional TR4 can increase stress resistance. These results suggest that this new identified oxidative stress-FOXO3a-TR4 pathway is a fundamentally important mechanism regulating stress resistance and cell survival. PMID- 18388196 TI - Low-frequency electro-acupuncture and physical exercise improve metabolic disturbances and modulate gene expression in adipose tissue in rats with dihydrotestosterone-induced polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a complex endocrine and metabolic disorder associated with ovulatory dysfunction, hyperandrogenism, abdominal obesity, and insulin resistance. Pharmacotherapy is often unsatisfactory. This study evaluates the effects of low-frequency electro-acupuncture (EA) and physical exercise on metabolic disturbances and adipose tissue mRNA expression of selected genes in a rat PCOS model characterized by insulin resistance and adiposity. Dihydrotestosterone (inducing PCOS) or vehicle (control) was administrated continuously, beginning before puberty. At age 10 wk, PCOS rats were randomly divided into three groups; PCOS, PCOS EA, and PCOS exercise. PCOS EA rats received 2-Hz EA (evoking muscle twitches) three times/wk during 4-5 wk. PCOS exercise rats had free access to a running wheel for 4-5 wk. EA and exercise improved insulin sensitivity, measured by clamp, in PCOS rats. Exercise also reduced adiposity, visceral adipocyte size, and plasma leptin. EA increased plasma IGF-I. Real-time RT-PCR revealed increased expression of leptin and IL-6 and decreased expression of uncoupling protein 2 in visceral adipose tissue of PCOS rats compared with controls. EA restored the expression of leptin and uncoupling protein 2, whereas exercise normalized adipose tissue leptin and IL-6 expression in PCOS rats. Thus, EA and exercise ameliorate insulin resistance in rats with PCOS. This effect may involve regulation of adipose tissue metabolism and production because EA and exercise each partly restore divergent adipose tissue gene expression associated with insulin resistance, obesity, and inflammation. In contrast to exercise, EA improves insulin sensitivity and modulates adipose tissue gene expression without influencing adipose tissue mass and cellularity. PMID- 18388195 TI - Estrogen elicits dorsal root ganglion axon sprouting via a renin-angiotensin system. AB - Many painful conditions occur more frequently in women, and estrogen is a predisposing factor. Estrogen may contribute to some pain syndromes by enhancing axon outgrowth by sensory dorsal root ganglion (DRG) neurons. The objective of the present study was to define mechanisms by which estrogen elicits axon sprouting. The estrogen receptor-alpha agonist propyl pyrazole triol induced neurite outgrowth from cultured neonatal DRG neurons, whereas the estrogen receptor-beta agonist diarylpropionitrile was ineffective. 17beta-Estradiol (E2) elicited sprouting from peripherin-positive unmyelinated neurons, but not larger NF200-positive myelinated neurons. Microarray analysis showed that E2 up regulates angiotensin II (ANGII) receptor type 2 (AT2) mRNA in vitro, and studies in adult rats confirmed increased DRG mRNA and protein in vivo. AT2 plays a central role in E2-induced axon sprouting because AT2 blockade by PD123,319 eliminated estrogen-mediated sprouting in vitro. We assessed whether AT2 may be responding to locally synthesized ANGII. DRG from adult rats expressed mRNA for renin, angiotensinogen, and angiotensin converting enzyme (ACE), and protein products were present and occasionally colocalized within neurons and other DRG cells. We determined if locally synthesized ANGII plays a role in estrogen mediated sprouting by blocking its formation using the ACE inhibitor enalapril. ACE inhibition prevented estrogen-induced neuritogenesis. These findings support the hypothesis that estrogen promotes DRG nociceptor axon sprouting by up regulating the AT2 receptor, and that locally synthesized ANGII can induce axon formation. Therefore, estrogen may contribute to some pain syndromes by enhancing the pro-neuritogenic effects of AT2 activation by ANGII. PMID- 18388197 TI - Autoimmune hypophysitis of SJL mice: clinical insights from a new animal model. AB - Autoimmune hypophysitis (AH) is a rare but increasingly recognized disease of the pituitary gland. Its autoantigens are unknown, and the management is difficult because it is often misdiagnosed as a nonsecreting adenoma. By immunizing female SJL/J mice with mouse pituitary extracts, we established a new mouse model of experimental AH. Immunized mice developed severe lymphocytic infiltration in the anterior pituitary that closely mimicked the human pathology. In the early phase of experimental AH, the pituitary enlarged, consistent with the compression symptoms reported by hypophysitis patients at presentation. In the florid phase, adrenal insufficiency and pituitary antibodies developed, in strong correlation with the pituitary pathology. In the late phase, hypothyroidism ensued, and the pituitary gland became atrophic. Using immune sera as probes in a two-dimensional immunoblotting screen followed by mass spectrometry, we identified several proteins that could function as pituitary autoantigens. These findings provide new insights into the pathogenesis of AH, and establish a platform for developing novel diagnostic biomarkers and therapeutics. PMID- 18388198 TI - Inhibition of vascular endothelial growth factor in the primate ovary up regulates hypoxia-inducible factor-1alpha in the follicle and corpus luteum. AB - Vascular endothelial growth factor (VEGF)-dependent angiogenesis is crucial for follicular growth, and corpus luteum formation and function, in the primate ovary. In the ovary VEGF can be hormonally regulated, but in other systems, the main regulator of VEGF expression is hypoxia. We hypothesized that hypoxia was involved in the regulation of angiogenesis in the cycling ovary. We therefore used immunohistochemistry to localize hypoxia-inducible factor (HIF)-1alpha in the marmoset ovary across the ovarian cycle. We also investigated the effect of VEGF inhibition, using VEGF Trap (aflibercept), on HIF-1alpha localization during the follicular and luteal phases of the cycle. Finally, we studied the effect of chorionic gonadotropin stimulation of the corpus luteum during early pregnancy. Nuclear HIF-1alpha staining was largely absent from normally growing preantral and antral follicles. However, there was marked up-regulation of nuclear HIF 1alpha in the granulosa cells at ovulation that persisted into the early corpus luteum. Mature corpora lutea and those collected during early pregnancy had minimal nuclear HIF-1alpha staining. The inhibition of VEGF in the mid-luteal stage resulted in a time-dependent up-regulation of luteal nuclear HIF-1alpha staining (P < 0.05). There was never any nuclear HIF-1alpha in the theca cells of the follicle, but VEGF Trap treatment during the follicular (P < 0.001) or luteal (P < 0.001) phase increased the proportion of antral follicles with nuclear HIF 1alpha staining in the granulosa cells. These results indicate that HIF-1alpha is up-regulated after vascular inhibition, using VEGF Trap, in the follicle and corpus luteum. However, it is also acutely up-regulated during ovulation. This suggests a role for HIF-1alpha in both hypoxic and hormonal regulation of ovarian VEGF expression in vivo. PMID- 18388199 TI - New enzymes involved in the mechanism of action of epidermal growth factor in a clonal strain of Leydig tumor cells. AB - The studies presented herein were designed to investigate the effect of mouse epidermal growth factor (mEGF) on arachidonic acid (AA) release in a clonal strain of cultured murine Leydig cells (designed MA-10). In MA-10 cells, mEGF promotes AA release and metabolism to lipoxygenated products to induce the steroidogenic acute regulatory (StAR) protein. However, the mechanism by which mEGF releases AA in these cells is not totally elucidated. We show that mEGF produces an increment in the mitochondrial AA content in a short-term incubation (30 min). This AA is released by the action of a mitochondrial acyl-CoA thioesterase (Acot2), as demonstrated in experiments in which Acot2 was down or overexpressed. This AA in turn regulates the StAR protein expression, indirect evidence of its metabolism to lipoxygenated products. We also show that mEGF induces the expression (mRNA and protein) of Acot2 and an acyl-CoA synthetase that provides the substrate, arachidonyl-CoA, to Acot2. This effect is also observed in another steroidogenic cell line, the adrenocortical Y1 cells. Taken together, our results show that: 1) mEGF can induce the generation of AA in a specific compartment of the cells, i.e. the mitochondria; 2) mEGF can up-regulate acyl-CoA synthetase and Acot2 mRNA and protein levels; and 3) mEGF-stimulated intramitochondrial AA release leads to StAR protein induction. PMID- 18388200 TI - Reduced plasma high-density lipoprotein cholesterol in hyperthyroid mice coincides with decreased hepatic adenosine 5'-triphosphate-binding cassette transporter 1 expression. AB - The aim of the study was to investigate the influence of severe hyperthyroidism on plasma high-density lipoprotein cholesterol (HDL-C). Recently, it was shown in mice that increasing doses of T(3) up-regulate hepatic expression of scavenger receptor class B, type I, resulting in increased clearance of plasma HDL-C. Here, we show that severe hyperthyroidism in mice did not affect hepatic expression of scavenger receptor class B, type I, but reduced hepatic expression of ATP-binding cassette transporter 1, accompanied by a 40% reduction of HDL-C. The sterol content of bile, liver, and feces was markedly increased, accompanied by up regulation of hepatic cholesterol 7alpha-hydroxylase, and ATP-binding cassette transporter 5, which is known to promote biliary sterol secretion upon dimerization with ATP-binding cassette transporter 8. Both control and hyperthyroid mice exerted identical plasma clearance of iv injected [(3)H]HDL-C, supporting the view that severe hyperthyroidism does not affect HDL-C clearance but, rather, its formation via hepatic ATP-binding cassette transporter 1. PMID- 18388201 TI - The lattice as allosteric effector: structural studies of alphabeta- and gamma tubulin clarify the role of GTP in microtubule assembly. AB - GTP-dependent microtubule polymerization dynamics are required for cell division and are accompanied by domain rearrangements in the polymerizing subunit, alphabeta-tubulin. Two opposing models describe the role of GTP and its relationship to conformational change in alphabeta-tubulin. The allosteric model posits that unpolymerized alphabeta-tubulin adopts a more polymerization competent conformation upon GTP binding. The lattice model posits that conformational changes occur only upon recruitment into the growing lattice. Published data support a lattice model, but are largely indirect and so the allosteric model has prevailed. We present two independent solution probes of the conformation of alphabeta-tubulin, the 2.3 A crystal structure of gamma-tubulin bound to GDP, and kinetic simulations to interpret the functional consequences of the structural data. These results (with our previous gamma-tubulin:GTPgammaS structure) support the lattice model by demonstrating that major domain rearrangements do not occur in eukaryotic tubulins in response to GTP binding, and that the unpolymerized conformation of alphabeta-tubulin differs significantly from the polymerized one. Thus, geometric constraints of lateral self-assembly must drive alphabeta-tubulin conformational changes, whereas GTP plays a secondary role to tune the strength of longitudinal contacts within the microtubule lattice. alphabeta-Tubulin behaves like a bent spring, resisting straightening until forced to do so by GTP-mediated interactions with the growing microtubule. Kinetic simulations demonstrate that resistance to straightening opposes microtubule initiation by specifically destabilizing early assembly intermediates that are especially sensitive to the strength of lateral interactions. These data provide new insights into the molecular origins of dynamic microtubule behavior. PMID- 18388203 TI - Crystal structure and functional analysis of tetracenomycin ARO/CYC: implications for cyclization specificity of aromatic polyketides. AB - Polyketides are a class of natural products with highly diverse chemical structures and pharmaceutical activities. Polyketide cyclization, promoted by the aromatase/cyclase (ARO/CYC), helps diversify aromatic polyketides. How the ARO/CYC promotes highly specific cyclization is not well understood because of the lack of a first-ring ARO/CYC structure. The 1.9 A crystal structure of Tcm ARO/CYC reveals that the enzyme belongs to the Bet v1-like superfamily (or STAR domain family) with a helix-grip fold, and contains a highly conserved interior pocket. Docking, mutagenesis, and an in vivo assay show that the size, shape, and composition of the pocket are important to orient and specifically fold the polyketide chain for C9-C14 first-ring and C7-C16 second-ring cyclizations. Two pocket residues, R69 and Y35, were found to be essential for promoting first- and second-ring cyclization specificity. Different pocket residue mutations affected the polyketide product distribution. A mechanism is proposed based on the structure-mutation-docking results. These results strongly suggest that the regiospecific cyclizations of the first two rings and subsequent aromatizations take place in the interior pocket. The chemical insights gleaned from this work pave the foundation toward defining the molecular rules for the ARO/CYC cyclization specificity, whose rational control will be important for future endeavors in the engineered biosynthesis of novel anticancer and antibiotic aromatic polyketides. PMID- 18388204 TI - Predictors of blood pressure fall with continuous positive airway pressure (CPAP) treatment of obstructive sleep apnoea (OSA). AB - BACKGROUND: Obstructive sleep apnoea (OSA) is associated with high cardiovascular morbidity and mortality. Randomised controlled trials have shown that, on average, treatment of OSA with continuous positive airway pressure (CPAP) reduces blood pressure (BP) by 3-5 mm Hg, although with considerable variation between individuals. No predictors of the change in BP with CPAP have been convincingly identified. This prospective study aimed to determine predictors of BP change, which might provide an insight into the aetiology of the raised BP seen in untreated OSA. METHODS: Eighty-six patients with daytime hypersomnolence warranting treatment with CPAP were recruited. 24 h mean BP (24 hMBP), subjective sleepiness, fasting venous blood samples and anthropometric measurements were assessed at baseline and after 6 months of CPAP treatment. RESULTS: The mean (SD) 24 hMBP fell at 6 months from 101.0 (10.3) mm Hg to 96.1 (9.1) mm Hg (change 4.92 mm Hg (95% CI -2.8 to -7.1)). The Epworth Sleepiness Score (ESS) fell from a median of 16 (IQR 12-18) to 4 (2-7) with a mean fall of 9.7 (95% CI 8.6 to 10.8). Several factors correlated with the fall in 24 hMBP but, after allowing for the baseline 24 hMBP, only the fall in ESS and the body mass index (BMI) remained significant independent predictors (p = 0.006 and 0.007, respectively). There was also a correlation between the fall in 24 hMBP and the fall in pulse rate (r = 0.44, p<0.001). Baseline severity of OSA, overnight hypoxia, caffeine intake or being on antihypertensive drugs were not independent predictors of a fall in 24 hMBP. CONCLUSION: Improvement in hypersomnolence and the BMI are independent correlates of the fall in 24 hMBP following CPAP therapy. Markers of initial OSA severity did not predict the fall in 24 hMBP. This suggests that sleep fragmentation and its effects may be more important than hypoxia in the pathogenesis of the hypertension associated with human sleep apnoea. PMID- 18388205 TI - Ambulatory blood pressure in children with obstructive sleep apnoea: a community based study. AB - BACKGROUND: Childhood obstructive sleep apnoea (OSA) is increasingly being recognised. Its effects on blood pressure (BP) elevation and hypertension are still controversial. OBJECTIVE: To evaluate the association between OSA and ambulatory BP in children. METHODS: Children aged 6-13 years from randomly selected schools were invited to undergo overnight sleep study and ambulatory BP monitoring after completing a validated OSA questionnaire. OSA was diagnosed if the obstructive apnoea-hypopnoea index (AHI) was >1, and normal controls had AHI <1 and snoring <3 nights per week. Children with OSA were subdivided into a mild group (AHI 1-5) and moderate to severe group (AHI >5). RESULTS: 306 subjects had valid sleep and daytime BP data. Children with OSA had significantly higher BP than normal healthy children during both sleep and wakefulness. BP levels increased with the severity of OSA, and children with moderate to severe disease (AHI >5) were at significantly higher risk for nocturnal systolic (OR 3.9 (95% CI 1.4 to 10.5)) and diastolic (OR 3.3 (95% CI 1.4 to 8.1)) hypertension. Multiple linear regression revealed a significant association between oxygen desaturation index and AHI with daytime and nocturnal BP, respectively, independent of obesity. CONCLUSIONS: OSA was associated with elevated daytime and nocturnal BP, and is an independent predictor of nocturnal hypertension. This has important clinical implications as childhood elevated BP predicts future cardiovascular risks. Future studies should examine the effect of therapy for OSA on changes in BP. PMID- 18388202 TI - Oxygen-regulated isoforms of cytochrome c oxidase have differential effects on its nitric oxide production and on hypoxic signaling. AB - Recently, it has been reported that mitochondria possess a novel pathway for nitric oxide (NO) synthesis. This pathway is induced when cells experience hypoxia, is nitrite (NO(2)(-))-dependent, is independent of NO synthases, and is catalyzed by cytochrome c oxidase (Cco). It has been proposed that this mitochondrially produced NO is a component of hypoxic signaling and the induction of nuclear hypoxic genes. In this study, we examine the NO(2)(-)-dependent NO production in yeast engineered to contain alternative isoforms, Va or Vb, of Cco subunit V. Previous studies have shown that these isoforms have differential effects on oxygen reduction by Cco, and that their genes (COX5a and COX5b, respectively) are inversely regulated by oxygen. Here, we find that the Vb isozyme has a higher turnover rate for NO production than the Va isozyme and that the Vb isozyme produces NO at much higher oxygen concentrations than the Va isozyme. We have also found that the hypoxic genes CYC7 and OLE1 are induced to higher levels in a strain carrying the Vb isozyme than in a strain carrying the Va isozyme. Together, these results demonstrate that the subunit V isoforms have differential effects on NO(2)(-)-dependent NO production by Cco and provide further support for a role of Cco in hypoxic signaling. These findings also suggest a positive feedback mechanism in which mitochondrially produced NO induces expression of COX5b, whose protein product then functions to enhance the ability of Cco to produce NO in hypoxic/anoxic cells. PMID- 18388206 TI - Incidence and predictive factors of lower respiratory tract infections among the very elderly in the general population. The Leiden 85-plus Study. AB - OBJECTIVES: To target preventive strategies in old age, which of the very elderly are predisposed to developing lower respiratory tract infections was investigated. DESIGN: Prospective observational follow-up study. SETTING: General population. PARTICIPANTS: Unselected cohort of 587 participants aged 85 years in Leiden, The Netherlands. MEASUREMENTS: As reported in the literature, predictive factors were selected and assessed at baseline. During a 5 year follow-up period, information on the development of lower respiratory tract infections was obtained from general practitioners or nursing home physicians. Associations between predictive factors were analysed with Cox regression, and population attributable risks were calculated. RESULTS: The incidence of lower respiratory tract infections among persons aged 85-90 years was 94 (95% CI 80-108) per 1000 person years. After multivariate analysis, history of chronic obstructive pulmonary disease (COPD), smoking, oral glucocorticosteroid use, severe cognitive impairment, history of stroke and declined functional status remained independently associated with the occurrence of lower respiratory tract infections. Smoking was the greatest contributor with a population attributable risk of 32%. CONCLUSION: In the very old, smoking, COPD, stroke and declined functional status were associated with the occurrence of lower respiratory tract infections and provide a means of targeting patients at risk of severe health complications. PMID- 18388207 TI - Re: Spinal epidural hemangiomas: various types of MR imaging features with histopathologic correlation. PMID- 18388208 TI - Response to the commentary "how do we spin wingspan?". PMID- 18388209 TI - Re: Turk et al and the "how do we spin wingspan?" commentary. PMID- 18388210 TI - Endovascular treatment of giant serpentine aneurysms. AB - Giant serpentine aneurysms are fusiform partially thrombosed aneurysms with a separate outflow tract to normal distal cerebral vessels. Three patients with giant serpentine aneurysms of the anterior and middle cerebral arteries were treated with endovascular occlusion of the aneurysmal lumen with coils or glue after balloon test occlusion of the involved vessel. In all 3 patients, leptomeningeal collateral circulation was sufficient to prevent distal ischemia. PMID- 18388211 TI - MR imaging-based decision in thrombolytic therapy for stroke on awakening: report of 2 cases. AB - Patients with stroke on awakening are denied the potential benefit of thrombolysis on the grounds that the onset time is unknown. Relying on clinical and MR imaging to indicate the most appropriate treatment could be more rational. We report 2 cases of stroke with unknown onset time. In both cases, anamnesis and MR imaging indicated that we might still be within 6 hours from stroke onset, with salvageable tissue. Arterial recanalization was successfully performed in both cases. PMID- 18388212 TI - Can MR imaging diagnose adult-onset Alexander disease? AB - BACKGROUND AND PURPOSE: In recent years, the discovery that mutations in the glial fibrillary acidic protein gene (GFAP) were responsible for Alexander disease (AD) brought recognition of adult cases. The purpose of this study was to demonstrate that MR imaging allows identification of cases of AD with adult onset (AOAD), which are remarkably different from infantile cases. MATERIALS AND METHODS: In this retrospective study, brain and spinal cord MR imaging studies of 11 patients with AOAD (7 men, 4 women; age range, 26-64 years; mean age, 43.6 years), all but 1 genetically confirmed, were reviewed. Diffusion and spectroscopic investigations were available in 6 patients each. RESULTS: Atrophy and changes in signal intensity in the medulla oblongata and upper cervical spinal cord were present in 11 of 11 cases and were the diagnostic features of AOAD. Minimal to moderate supratentorial periventricular abnormalities were seen in 8 patients but were absent in the 3 oldest patients. In these patients, postcontrast enhancement was also absent. Mean diffusivity was not altered except in abnormal white matter (WM). Increase in myo-inositol (mIns) was also restricted to abnormal periventricular WM. CONCLUSIONS: Awareness of the MR pattern described allows an effective selection of the patients who need genetic investigations for the GFAP gene. This MR pattern even led to identification of asymptomatic cases and should be regarded as highly characteristic of AOAD. PMID- 18388213 TI - The frontal intersinus septal air cell: a new hypothesis of its origin. AB - BACKGROUND AND PURPOSE: Air cells are often seen within the frontal intersinus septum. These cells have traditionally been thought to arise from displaced ethmoid cells from the frontal recess. This study explores the possibility that such cells may actually be diverticula from the frontal sinuses themselves and not of a direct ethmoid origin. MATERIALS AND METHODS: A prospective study of 200 consecutive CT scans in the coronal and axial planes was performed on patients without a history of recent trauma. The images were interpreted independently by a radiologist and an otolaryngologist. The CT studies were evaluated for the presence of a central intersinus septal air cell. If such a cell was identified, it was further classified as either being completely isolated from both frontal sinuses by a bony rim or as a communicating diverticulum from one of the frontal sinuses. If a central cell was present, it was also assessed for how much of the height of the intersinus septum it involved (lower one-half or full height). RESULTS: There was a complete concordance of the results between the 2 observers. An intersinus septal air cell was seen in 61 (30.5%) of the 200 cases, and 85.3% of these cells were clearly seen to communicate anteromedially with either one of the frontal sinuses or both frontal sinuses (3 cases). In 9 (4.5%) of the 200 cases, the central cell had no demonstrable connection to either frontal sinus. Of the 61 cases with a central cell, 55 (90.16%) of the cells occupied the full height of the septum, and 6 (9.84%) only involved the lower half of the septum. CONCLUSION: Contrary to the present convention that frontal intersinus septal cells originate as displaced ethmoid cells from the frontal recess, we found that most such cells are actually diverticula from the frontal sinuses themselves. PMID- 18388214 TI - Interval change in size of venous pouch canine bifurcation aneurysms over a 10 month period. AB - BACKGROUND AND PURPOSE: The natural history study of experimental aneurysms is important for the evaluation of new endovascular occlusion devices. Our purpose was to evaluate the natural history of experimental venous pouch bifurcation aneurysms in mongrel dogs up to a 10-month period. MATERIALS AND METHODS: Serial digital subtraction angiography was performed in 5 bifurcation aneurysms 1, 4, 7, and 10 months after surgical creation. Aneurysm dimensions, including height, width, and neck diameter, and animal body weights were measured. Comparisons of each parameter were performed using the Friedman test and the paired Wilcoxon signed-rank test. RESULTS: Four of 5 aneurysms were patent during a 10-month follow-up period. One aneurysm was regarded as a partially thrombosed aneurysm at 1 month, though the extent of partial thrombosis lessened at 10 months. Bifurcation aneurysms progressively increased in size (aneurysm height, width, and neck diameter) during the first several months. CONCLUSION: If this experimental model is used to evaluate new endovascular devices for cerebral aneurysm treatment, investigators should be aware of early progressive aneurysm enlargement. PMID- 18388215 TI - Patient skin dose during neuroembolization by multiple-point measurement using a radiosensitive indicator. AB - BACKGROUND AND PURPOSE: Although neuroembolization has recently spread quickly, sufficient attention has not been focused on the associated radiation exposure. The purpose of this research was to evaluate the patient's entrance skin dose (ESD) during neuroembolizations in 6 institutions. MATERIALS AND METHODS: This study was approved by all of the 6 institutional review boards, and all of the patients gave informed consent. This study included a total of 103 consecutive neuroembolizations in the 6 institutions. Patient ESDs during the procedures were evaluated by using caps that had 44 radiosensitive indicators adherent to the surface. The patient ESDs were calculated from the color difference of the indicators. To check for effects on the scalp, clinical follow-up was performed at 1-2 days, 2 weeks, and 3 months after the procedure. RESULTS: The averages of total fluoroscopic time, total number of digital subtraction angiography frames, and dose area product were 67.1 +/- 41.6 minutes, 883 +/- 626, and 257 +/- 150 Gy x cm(2), respectively. The average maximum ESD for each patient was 1.9 +/- 1.1 Gy (range, 0.4-5.6 Gy; median, 1.5 Gy). The average maximum ESDs of each institution ranged from 1.0 to 2.4 Gy. Epilation was observed in 6 patients. CONCLUSIONS: The maximum ESDs during neuroembolizations exceed the thresholds for radiation skin injuries in some cases. PMID- 18388216 TI - Performance evaluation of radiologists with artificial neural network for differential diagnosis of intra-axial cerebral tumors on MR images. AB - BACKGROUND AND PURPOSE: Previous studies have suggested that use of an artificial neural network (ANN) system is beneficial for radiological diagnosis. Our purposes in this study were to construct an ANN for the differential diagnosis of intra-axial cerebral tumors on MR images and to evaluate the effect of ANN outputs on radiologists' diagnostic performance. MATERIALS AND METHODS: We collected MR images of 126 patients with intra-axial cerebral tumors (58 high grade gliomas, 37 low-grade gliomas, 19 metastatic tumors, and 12 malignant lymphomas). We constructed a single 3-layer feed-forward ANN with a Levenberg Marquardt algorithm. The ANN was designed to differentiate among 4 categories of tumors (high-grade gliomas, low-grade gliomas, metastases, and malignant lymphomas) with use of 2 clinical parameters and 13 radiologic findings in MR images. Subjective ratings for the 13 radiologic findings were provided independently by 2 attending radiologists. All 126 cases were used for training and testing of the ANN based on a leave-one-out-by-case method. In the observer test, MR images were viewed by 9 radiologists, first without and then with ANN outputs. Each radiologist's performance was evaluated through a receiver operating characteristic (ROC) analysis on a continuous rating scale. RESULTS: The averaged area under the ROC curve for ANN alone was 0.949. The diagnostic performance of the 9 radiologists increased from 0.899 to 0.946 (P < .001) when they used ANN outputs. CONCLUSIONS: The ANN can provide useful output as a second opinion to improve radiologists' diagnostic performance in the differential diagnosis of intra-axial cerebral tumors seen on MR imaging. PMID- 18388217 TI - State of the art: 3T imaging of the membranous labyrinth. AB - This article reviews the anatomy of the membranous labyrinth and demonstrates the ability of high-resolution MR imaging at 3T to visualize the neurosensory epithelium by using the latest fast spin-echo techniques. PMID- 18388218 TI - Metabolic assessment of gliomas using 11C-methionine, [18F] fluorodeoxyglucose, and 11C-choline positron-emission tomography. AB - BACKGROUND AND PURPOSE: Positron-emission tomography (PET) is a useful tool in oncology. The aim of this study was to assess the metabolic activity of gliomas using (11)C-methionine (MET), [(18)F] fluorodeoxyglucose (FDG), and (11)C-choline (CHO) PET and to explore the correlation between the metabolic activity and histopathologic features. MATERIALS AND METHODS: PET examinations were performed for 95 primary gliomas (37 grade II, 37 grade III, and 21 grade IV). We measured the tumor/normal brain uptake ratio (T/N ratio) on each PET and investigated the correlations among the tracer uptake, tumor grade, tumor type, and tumor proliferation activity. In addition, we compared the ease of visual evaluation for tumor detection. RESULTS: All 3 of the tracers showed positive correlations with astrocytic tumor (AT) grades (II/IV and III/IV). The MET T/N ratio of oligodendroglial tumors (OTs) was significantly higher than that of ATs of the same grade. The CHO T/N ratio showed a significant positive correlation with histopathologic grade in OTs. Tumor grade and type influenced MET uptake only. MET T/N ratios of more than 2.0 were seen in 87% of all of the gliomas. All of the tracers showed significantly positive correlations with Mib-1 labeling index in ATs but not in OTs and oligoastrocytic tumors. CONCLUSION: MET PET appears to be useful in evaluating grade, type, and proliferative activity of ATs. CHO PET may be useful in evaluating the potential malignancy of OTs. In terms of visual evaluation of tumor localization, MET PET is superior to FDG and CHO PET in all of the gliomas, due to its straightforward detection of "hot lesions". PMID- 18388219 TI - Prevalence and evolution of intracranial hemorrhage in asymptomatic term infants. AB - BACKGROUND AND PURPOSE: Subdural hemorrhage (SDH) is often associated with infants experiencing nonaccidental injury (NAI). A study of the appearance and natural evolution of these birth-related hemorrhages, particularly SDH, is important in the forensic evaluation of NAI. The purpose of this study was to determine the normal incidence, size, distribution, and natural history of SDH in asymptomatic term neonates as detected by sonography (US) and MR imaging within 72 hours of birth. MATERIALS AND METHODS: Birth history, delivery method, duration of each stage of labor, pharmaceutic augmentation, and complications during delivery as well as postnatal physical examination were recorded. Brain MR imaging and US were performed on 101 asymptomatic term infants at 3-7 days, 2 weeks, 1 month, and 3 months. Clinical follow-up at 24 months was recorded. RESULTS: Forty-six neonates had SDH by MR imaging within 72 hours of delivery. SDH was seen in both vaginal and cesarean deliveries. All neonates were asymptomatic, with normal findings on physical examination. All 46 had supratentorial SDH seen in the posterior cranium. Twenty (43%) also had infratentorial SDH. US detected 11 of the 20 (55%) infratentorial SDHs and no supratentorial SDH. Most SDHs present at birth were or =65 years. CONCLUSIONS: Burn prevention efforts are geared towards increasing smoke alarm ownership and improving maintenance of functional status. Results suggest that the selective use of photoelectric alarms by fire injury prevention programs or consumers may provide longer-term protection in similar populations. Designing smoke alarms that minimize nuisance alarming may also result in longer term functionality. PMID- 18388227 TI - National study of physical and sexual assault among women with disabilities. AB - OBJECTIVE: To examine the association between the level of disability impairment and physical and sexual assault in a sample of US women at least 18 years of age. DESIGN, SETTING AND PARTICIPANTS: Retrospective longitudinal study of 6273 non institutionalized US women from 8000 women participating in the 1995-1996 National Violence Against Women (NVAW) Survey. MAIN OUTCOME MEASURE: Women's experiences of physical and sexual assault in the 12 months before the NVAW interview. RESULTS: Most women reported having no disability (n = 5008, 79.8%) and/or not experiencing an assault in the year before their interview (n = 6018, 95.9%). Less than 5% (n = 280) reported having a disability that severely limited daily activities, and 15.7% (n = 985) reported having a disability that moderately limited activities. Less than 4% (n = 218) of the women reported a physical-only assault, and less than 1% (n = 37) reported being sexually assaulted. Women with severe disability impairments were four times more likely to be sexually assaulted than women with no reported disabilities (RR = 4.0, 95% CI 1.5 to 10.6). Little difference in the risk of sexual assault was found between women with moderate disability impairments and those reporting no disabilities (RR = 1.0, 95% CI 0.3 to 2.8). Women with severe (RR = 1.6, 95% CI 0.9 to 3.0) and moderate (RR = 1.2, 95% CI 0.8 to 1.9) disability impairments were at greater risk, although not quite significantly so, of physical-only assault than were women without a disability. CONCLUSION: The findings suggest that women with disabilities that severely limit activities of daily living are at increased risk of sexual assault. PMID- 18388228 TI - Using online databases to find journal articles on injury prevention and safety promotion research: key journals and the databases that index them. AB - OBJECTIVES: (1) To determine the trends of the number of journals that publish injury prevention and safety promotion (IPSP) articles and the number of articles published each year from 1900 through 2006; (2) to determine the coverage of key IPSP journals in widely used literature databases. METHODS: Journals were identified through a structured procedure that included hand-searching selected classification categories of two major listings of periodicals that publish four or more IPSP-relevant articles per year. Article relevance was assessed using the inclusion criteria for SafetyLit (a database of scholarly literature selected for its relevance to the IPSP field). Each identified journal was hand-searched from its first issue, and IPSP-relevant articles were added to the SafetyLit archive database. A MySQL database was used to perform basic queries and statistical summary analyses for journals published 1900-2006. The number of IPSP journals and journal articles was plotted for these years to identify publication trends. The publication year range of each IPSP journal was compared with the coverage, if any, of the journals in each of the five databases. RESULTS: Of the 17 839 journals assessed, 597 met the threshold for inclusion in this study. This amounts to thousands of IPSP-related articles published each year: 6100 in 2006. Only 160 (26.8%) of the journals are included in all five databases, and none have all publication years included in all of the databases. CONCLUSIONS: Some key IPSP journals are completely excluded from some of the databases. Thus, a search in a single database may miss key information from one of the many disciplines that publish IPSP-relevant information. PMID- 18388229 TI - Clustered and missing data in the US National Trauma Data Bank: implications for analysis. AB - BACKGROUND: Injury researchers are increasingly using the US National Trauma Data Bank (NTDB). However, there are some methodological issues that might threaten the validity of studies that use this database for injury research. METHODS: Two methodological issues were evaluated: clustering of patients within trauma centers and missing data. To illustrate how these issues might affect the results of a study, the following four analytical approaches that evaluated the association between patients' blood alcohol concentration (BAC) in the emergency department (ED), patients' resource utilization, and ED or hospital disposition were compared: (A) deleting subjects with missing BAC and ignoring clustering of patients within trauma centers; (B) deleting subjects with missing BAC while taking into account clustering; (C) using imputed values for patients' BAC and ignoring the clustering issue; (D) using the imputed data while taking into account clustering. RESULTS: Adjustment for clustering of patients within trauma centers increased the CIs in models B and D. The results of the analyses based on imputed data showed that estimates based on complete case analysis were biased. For example, the odds ratio for the use of a head CT scan fell from 1.84 (95% CI 1.49 to 2.28) in approach B to 1.26 (95% CI 0.98 to 1.64) in approach D. CONCLUSIONS: Excluding patients with missing values for BAC in studies that evaluate the association between this variable and patients' resource utilization and ED or hospital disposition, using the NTDB, led to biased estimates. Furthermore, ignoring the clustering design led to artificially narrow CIs. PMID- 18388230 TI - Examining the burden of work-related hospitalized injuries: definitional issues. AB - BACKGROUND: Identifying work-related injuries from hospitalization data is not straightforward. How a work-related injury, either acute or non-acute, is defined will affect injury enumeration, injury burden estimates, and subsequent priority setting for prevention activity. AIM: To examine the effect of different case identification criteria on the number and rate of hospitalized work-related injuries and to identify the type of pathologies that are recorded for work related admissions that have an external cause code. METHOD: Identified work related hospitalization admissions from 1 July 2000 to 30 June 2005 were analysed according to different injury case-selection criteria (ie, a principal diagnosis in the ICD-10-AM range S00-T98 and/or an external cause code in the ICD-10-AM range V01-Y98) and different admission restrictions (ie, exclusion of: transfers and type changes; re-admissions within 28 days; or day-only admissions). The principal diagnosis recorded for work-related admissions that had an external cause code were examined. RESULTS: The incidence of identified work-related hospitalized injuries varied widely, from 289 to 611 per 100,000 workers depending on the identification criteria adopted. The exclusion of day-only stays reduced the identified work-related injury admissions by about one-third for each definitional condition. Where only an external cause code was used to identify an injury, the principal diagnoses codes were predominantly injuries, poisoning, and certain other consequences of external causes (74.6%) and diseases of the musculoskeletal system and connective tissue (15.5%). CONCLUSIONS: Case-selection criteria adopted to identify work-related hospitalized injuries should be carefully considered. It is recommended that inclusion of certain musculoskeletal conditions that are likely to arise from repetitive minor trauma over time (ie non-acute injuries) should be considered in calculating the estimate of the burden of all work-related hospitalized injuries. PMID- 18388231 TI - Feasibility of standardized injury surveillance and reporting: a comparison of data from four Asian nations. AB - BACKGROUND: To address the increasing number of injuries in developing countries, the World Health Organization (WHO) encourages the establishment of hospital based surveillance systems and systematic data collection. Although a computerized system is preferable in terms of efficiency, many developing countries have difficulty accessing the appropriate resources. OBJECTIVES: To assess the possibility of comparing and sharing data among countries, and then to discuss the possibility of establishing an international data management system through the internet. METHODS: A point-by-point comparison of data directories from injury surveillance systems in Thailand, Cambodia, Sri Lanka, and Japan was conducted using guidelines published by WHO as the standard. RESULTS: Thailand, Cambodia, and Sri Lanka used data items that are comparable to each other and to WHO guidelines, with few, readily amenable, differences. The Japanese system used quite different data items. CONCLUSIONS: Data comparability suggests the feasibility of a global data management system that can store data from various countries. Such a system, if made accessible over the internet, would benefit resource-constrained countries by providing them with a ready-made framework to implement a surveillance system at low cost. PMID- 18388232 TI - Effectiveness of multifaceted fall-prevention programs for the elderly in residential care. AB - BACKGROUND: Unintentional falls are particularly prevalent among older people and constitute a public health concern. Not much is known about the implications of multifaceted intervention programs implemented in residential care settings. OBJECTIVES: To evaluate the effectiveness of multifaceted intervention programs in reducing the number of falls, fallers, recurrent fallers, and injurious falls among older people living in residential care facilities. SEARCH STRATEGY: Comprehensive searches of Medline, PubMed, and EMBASE up to July 2007, the cited literature lists of each included study, and the internet engines Google Scholar, Yahoo, and Dogpile were performed to identify eligible studies. SELECTION CRITERIA: Eligible studies for this review were those that had randomized, controlled trials with adequate follow-up study components in their design. Studies that included elderly people in residential care who participated in multifaceted falls-prevention programs were included. DATA COLLECTION AND ANALYSIS: Two authors independently extracted the necessary data. Studies were assessed for quality by the criteria of Downs and Black. The results of the included studies have been reviewed narratively. MAIN RESULTS: From 21 articles potentially relevant to the topic, five studies met the inclusion criteria and all were reasonably well conducted. Three reported significant reductions in the number of recurrent fallers, two reported significant reductions in the number of falls, and one reported significant reductions in the number of fallers. One other reported a reduction in the number of injurious falls in those who received the multifaceted prevention program compared with the control group. However, the analyses of this specific study were not based on intent-to-treat, so the effect of intervention on the number of injurious falls remains inconclusive. No study reported on adverse events, costs, or sustainability of the interventions. CONCLUSIONS: Multifaceted programs that encompass a wide range of intervention strategies have shown some evidence of efficacy. However, more well-designed research is required that assesses effects on injurious falls, quality of life, cost-effectiveness, and sustainability. PMID- 18388233 TI - Zero blood alcohol concentration limits for drivers under 21: lessons from Canada. AB - Graduated licensing programs (GLPs) that include zero or low blood alcohol concentration (BAC) restrictions have proven to be a popular and effective measure for improving traffic safety among young people. However, a major drawback of such programs, at least in Canada, is that the BAC restriction is lifted on completion of the GLP, which typically occurs around the age of 18 or 19. This corresponds to the legal drinking age in Canada, a time when alcohol consumption and rates of binge drinking increase. It is not surprising, then, that 18-20 year-old drivers are dramatically overrepresented in alcohol-related deaths and injuries. One way to address this problem is to raise the legal drinking age, as has occurred in the United States. In jurisdictions, like Canada, that are unlikely to raise the legal drinking age, other measures are necessary to separate drinking from driving among 18-20 year-olds. This article recommends that the zero BAC restrictions be extended beyond the completion of the GLP, until drivers reach the age of 21. The scientific evidence for such a measure is reviewed, and the growing government support for enacting such BAC limits in Canada is described. PMID- 18388234 TI - Injury control: an opportunity for China. AB - Injury control is an important health issue in China, but has been less well explored than in other countries. To encourage health policy makers to give greater attention to injury control, this report highlights the great damage caused by injuries and the neglected status of injury control in China. China's situation and the experiences and lessons from industrialized countries, especially the USA, are summarized. Finally, two opportunities to improve injury control in China are identified: establishment of a mechanism for multi department coordination and collaboration; and improvement of injury data surveillance. PMID- 18388235 TI - Unintentional pediatric submersion-injury-related hospitalizations in the United States, 2003. AB - The objective of this study was to examine the demographic characteristics and hospital resource utilization of submersion-injury-related hospitalizations among persons < or =20 years of age in the USA in 2003. All 1475 pediatric submersion injury-related hospital discharges in the Kids' Inpatient Database were identified by ICD-9-CM diagnosis code or external cause of injury code. These cases represent an estimated 2490 pediatric submersion-injury-related hospitalizations nationwide. Inpatient costs for these estimated hospitalizations were approximately $10 million. The overall pediatric submersion-injury-related rate of hospitalization was 3.0 per 100,000 persons. Children aged 0-4 years had the highest rate of hospitalization (7.7 per 100,000 persons). Children with permanent submersion-injury-related morbidity accounted for 5.8% of hospital admissions and 37.3% of hospital costs in our study, and children with submersion injury-related in-hospital death accounted for 11.6% of hospital admissions and 20.0% of hospital costs in our study. Prevention of submersion injury using focused, proven strategies deserves increased attention. PMID- 18388237 TI - Searching for studies for inclusion in Cochrane Systematic Reviews on injury prevention. PMID- 18388238 TI - Celebrating 15 years of global networking with the International Society of Child & Adolescent Injury Prevention. PMID- 18388239 TI - New injury and violence research website open to the world. PMID- 18388240 TI - New manual for estimating the economic costs of injuries due to interpersonal and self-directed violence. PMID- 18388243 TI - M3 muscarinic acetylcholine receptor-mediated signaling is regulated by distinct mechanisms. AB - We have used RNA interference previously to demonstrate that G protein-coupled receptor kinase 2 (GRK2) regulates endogenously expressed H1 histamine receptor in human embryonic kidney 293 cells. In this report, we investigate the regulation of endogenously expressed M(3) muscarinic acetylcholine receptor (M(3) mAChR). We show that knockdown of GRK2, GRK3, or GRK6, but not GRK5, significantly increased carbachol-mediated calcium mobilization. Stable expression of wild-type GRK2 or a kinase-dead mutant (GRK2-K220R) reduced calcium mobilization after receptor activation, whereas GRK2 mutants defective in Galpha(q) binding (GRK2-D110A, GRK2-R106A, and GRK2-R106A/K220R) had no effect on calcium signaling, suggesting that GRK2 primarily regulates G(q) after M(3) mAChR activation. The knockdown of arrestin-2 or arrestin-3 also significantly increased carbachol-mediated calcium mobilization. Knockdown of GRK2 and the arrestins also significantly enhanced carbachol-mediated activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), whereas prolonged ERK1/2 activation was only observed with GRK2 or arrestin-3 knockdown. We also investigated the role of casein kinase-1alpha (CK1alpha) and found that knockdown of CK1alpha increased calcium mobilization but not ERK activation. In summary, our data suggest that multiple proteins dynamically regulate M(3) mAChR-mediated calcium signaling, whereas GRK2 and arrestin-3 play the primary role in regulating ERK activation. PMID- 18388242 TI - Attenuation of experimental autoimmune hepatitis by exogenous and endogenous cannabinoids: involvement of regulatory T cells. AB - Immune-mediated liver diseases including autoimmune and viral hepatitis are a major health problem worldwide. Natural cannabinoids such as Delta(9) tetrahydrocannabinol (THC) effectively modulate immune cell function, and they have shown therapeutic potential in treating inflammatory diseases. We investigated the effects of THC in a murine model of concanavalin A (ConA) induced hepatitis. Intraperitoneal administration of THC after ConA challenge inhibited hepatitis as shown by significant decrease in liver enzymes and reduced liver tissue injury. Furthermore, THC treatment resulted in significant suppression of crucial inflammatory cytokines in ConA-induced hepatitis. It is noteworthy that THC treatment in ConA-injected mice led to significant increase in absolute number of Forkhead helix transcription factor p3+ T regulatory cells in liver. We were surprised to find that select cannabinoid receptor (CB1 or CB2) agonists were not able to block hepatitis either independently or in combination. However, CB1/CB2 mixed agonists were able to efficiently attenuate hepatitis similar to THC. The modulatory effect of THC in ConA-induced hepatitis was reversed by both CB1 and CB2 antagonists. We also observed that endogenous cannabinoid anandamide was able to reduce hepatitis by suppressing cytokine levels. In addition, deficiency or inhibition of endocannabinoid hydrolyzing enzyme fatty acid amide hydrolase (FAAH), which leads to increased levels of endogenous cannabinoids, resulted in decreased liver injury upon ConA challenge. Our data demonstrate that targeting cannabinoid receptors using exogenous or endogenous cannabinoids and use of FAAH inhibitors may constitute novel therapeutic modalities to treat immune-mediated liver inflammation. PMID- 18388245 TI - Beer and the respiratory muscles: the adverse effects of ascites. PMID- 18388244 TI - Roles of cytosolic phospholipase A2 and Src kinase in the early action of 2,3,7,8 tetrachlorodibenzo-p-dioxin through a nongenomic pathway in MCF10A cells. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, or dioxin) is known to induce rapid inflammatory cellular responses through the mechanism that has not yet been fully elucidated. In this report, we show that in MCF10A cells, an immortalized, normal mammary epithelial cell line, TCDD rapidly activates the enzymatic activity of cytosolic phospholipase A2 (cPLA2) as at-tested to by arachidonic acid release within 15 min, followed by activation of Src kinase and induction of several inflammation markers. Such an action of TCDD is clearly blocked by methylarachidonyl fluorophosphonate, a specific inhibitor of cPLA2, short interfering RNA against cPLA2, and several calcium signaling blockers, indicating that this action of TCDD is mediated by calcium-triggered activation of cPLA2. This action of TCDD is quite different from the classic action of TCDD to induce cytochrome P450 1A1 (CYP1A1) because blocking this newly identified pathway did not affect the induction of CYP1A1. Moreover, this newly identified pathway was found to depend only on aryl hydrocarbon receptor but not on aryl hydrocarbon receptor nuclear translocator. Together, these findings support the model that the early action of TCDD to induce rapid inflammatory responses is carried out through a characteristic "nongenomic" pathway, which is clearly different from the conventional model of action of TCDD through the "genomic" pathway. PMID- 18388247 TI - Effects of the laryngeal jet on nano- and microparticle transport and deposition in an approximate model of the upper tracheobronchial airways. AB - The extent to which laryngeal-induced flow features penetrate into the upper tracheobronchial (TB) airways and their related impact on particle transport and deposition are not well understood. The objective of this study was to evaluate the effects of including the laryngeal jet on the behavior and fate of inhaled aerosols in an approximate model of the upper TB region. The upper TB model was based on a simplified numerical reproduction of a replica cast geometry used in previous in vitro deposition experiments that extended to the sixth respiratory generation along some paths. Simulations with and without an approximate larynx were performed. Particle sizes ranging from 2.5 nm to 12 mum were considered using a well-tested Lagrangian tracking model. The model larynx was observed to significantly affect flow dynamics, including a laryngeal jet skewed toward the right wall of the trachea and a significant reverse flow in the left region of the trachea. Inclusion of the laryngeal model increased the tracheal deposition of nano- and micrometer particles by factors ranging from 2 to 10 and significantly reduced deposition in the first three bronchi of the model. Considering localized conditions, inclusion of the laryngeal approximation decreased deposition at the main carina and produced a maximum in local surface deposition density in the lobar-to-segmental bifurcations (G2-G3) for both 40-nm and 4-microm aerosols. These findings corroborate previous experiments and highlight the need to include a laryngeal representation in future computational and in vitro models of the TB region. PMID- 18388246 TI - Effect of long-term high-altitude hypoxia on fetal pulmonary vascular contractility. AB - Hypoxia in the fetus and/or newborn is associated with an increased risk of pulmonary hypertension. The present study tested the hypothesis that long-term high-altitude hypoxemia differentially regulates contractility of fetal pulmonary arteries (PA) and veins (PV) mediated by differences in endothelial NO synthase (eNOS). PA and PV were isolated from near-term fetuses of pregnant ewes maintained at sea level (300 m) or high altitude of 3,801 m for 110 days (arterial Po(2) of 60 Torr). Hypoxia had no effect on the medial wall thickness of pulmonary vessels and did not alter KCl-induced contractions. In PA, hypoxia significantly increased norepinephrine (NE)-induced contractions, which were not affected by eNOS inhibitor N(G)-nitro-l-arginine (l-NNA). In PV, hypoxia had no effect on NE-induced contractions in the absence of l-NNA. l-NNA significantly increased NE-induced contractions in both control and hypoxic PV. In the presence of l-NNA, NE-induced contractions of PV were significantly decreased in hypoxic lambs compared with normoxic animals. Acetylcholine caused relaxations of PV but not PA, and hypoxia significantly decreased both pD(2) and the maximal response of acetylcholine-induced relaxation in PV. Additionally, hypoxia significantly decreased the maximal response of sodium nitroprusside-induced relaxations of both PA and PV. eNOS was detected in the endothelium of both PA and PV, and eNOS protein levels were significantly higher in PV than in PA in normoxic lambs. Hypoxia had no significant effect on eNOS levels in either PA or PV. The results demonstrate heterogeneity of fetal pulmonary arteries and veins in response to long-term high-altitude hypoxia and suggest a likely common mechanism downstream of NO in fetal pulmonary vessel response to chronic hypoxia in utero. PMID- 18388248 TI - Sildenafil acts on the central nervous system increasing sympathetic activity. AB - Sildenafil induces vasodilation and is used for treating erectile dysfunction. Although its influence on resting heart function appears to be minimal, recent studies suggest that sildenafil can increase sympathetic activity. We therefore tested whether sildenafil injected into the central nervous system alters the autonomic control of the cardiovascular system in conscious rats. The effect of sildenafil citrate injected into the lateral cerebral ventricle was evaluated in conscious rats by means of the recording of lumbar sympathetic nerve activity (LSNA), spectral analysis of systolic arterial pressure and heart rate variability, spontaneous baroreflex sensitivity, and baroreflex control of LSNA. Intracerebroventricular (ICV, 100 microg /5 microl) administration of sildenafil caused remarkable tachycardia without significant change in basal arterial pressure and was associated with a conspicuous increase (47 +/- 14%) in LSNA. Spectral analysis demonstrated that systolic arterial pressure oscillations in the low frequency (LF) range were increased (from 6.3 +/- 1.5 to 12.8 +/- 3.8 mmHg(2)), whereas the high frequency (HF) range was not affected by ICV administration of sildenafil. Sildenafil increased pulse interval oscillations at LF and decreased them at HF. The LF-HF ratio increased from 0.04 +/- 0.01 to 0.17 +/- 0.06. Spontaneous baroreflex sensitivity measured by the sequence method and the baroreflex relationship between mean arterial pressure and LSNA were not affected by ICV administration of sildenafil. In conclusion, sildenafil elicited an increase in sympathetic nerve activity that is not baroreflex mediated, suggesting that this drug is able to elicit an autonomic imbalance of central origin. This finding may have implications for understanding the cardiovascular outcomes associated with the clinical use of this drug. PMID- 18388249 TI - Cytokine responses to acute and chronic exercise in multiple sclerosis. AB - Regular exercise reduces functional loss associated with multiple sclerosis (MS). However, the impact of exercise on inflammatory mediators associated with disease activity remains relatively unexplored. The purpose of this study was to determine whether ambulatory MS subjects would respond similarly to aerobic cycle training compared with matched controls on circulating immune variables, interleukin (IL)-6, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma. Eleven MS and 11 non-MS control subjects (8 women and 3 men in both groups) matched in age, height, body mass, body fat, and peak O(2) uptake completed the study. Subjects completed 30 min of cycle ergometry at 60% of peak O(2) uptake, 3 day/wk for 8 wk. Plasma cytokine concentrations were determined before and after exercise at weeks 0, 4, and 8. MS and control subjects showed a similar cytokine responses to exercise. IL-6 at rest tended to decrease (P = 0.08) with training in both groups. Resting plasma TNF-alpha tended to be higher in MS compared with controls throughout the study (P = 0.08). MS subjects showed elevated resting TNF alpha in MS at the end of the 8-wk program (P = 0.04), whereas resting TNF-alpha remained unchanged in controls (P > 0.05). Resting plasma IFN-gamma at rest was elevated in MS subjects (P = 0.008) and unchanged in controls at the end of the intervention (P > 0.05). The response of plasma IL-6, TNF-alpha, and IFN-gamma after a single bout of exercise was similar between MS and control subjects (P > 0.05). Additional research to understand the impact of exercise on immune variables in MS is warranted. PMID- 18388250 TI - Oral tolerance attenuates changes in in vitro lung tissue mechanics and extracellular matrix remodeling induced by chronic allergic inflammation in guinea pigs. AB - Recent studies emphasize the presence of alveolar tissue inflammation in asthma. Immunotherapy has been considered a possible therapeutic strategy for asthma, and its effect on lung tissue had not been previously investigated. Measurements of lung tissue resistance and elastance were obtained before and after both ovalbumin and acetylcholine challenges. Using morphometry, we assessed eosinophil and smooth muscle cell density, as well as collagen and elastic fiber content, in lung tissue from guinea pigs with chronic pulmonary allergic inflammation. Animals received seven inhalations of ovalbumin (1-5 mg/ml; OVA group) or saline (SAL group) during 4 wk. Oral tolerance (OT) was induced by offering ad libitum ovalbumin 2% in sterile drinking water starting with the 1st inhalation (OT1 group) or after the 4th (OT2 group). The ovalbumin-exposed animals presented an increase in baseline and in postchallenge resistance and elastance related to baseline, eosinophil density, and collagen and elastic fiber content in lung tissue compared with controls. Baseline and post-ovalbumin and acetylcholine elastance and resistance, eosinophil density, and collagen and elastic fiber content were attenuated in OT1 and OT2 groups compared with the OVA group. Our results show that inducing oral tolerance attenuates lung tissue mechanics, as well as eosinophilic inflammation and extracellular matrix remodeling induced by chronic inflammation. PMID- 18388251 TI - Discriminating between children with ADHD and classmates using peer variables. AB - OBJECTIVE: Impaired peer relationships have long been recognized as one of the major functional problems of children with ADHD, but no specific guidelines on clinical levels of impairment in this domain exist. METHOD: This study used Receiver Operating Characteristics methodology to determine what aspects of peer functioning best discriminate between children with ADHD and their classmates. Optimal cutoffs indicative of clinical levels of impairment associated with ADHD diagnosis were determined for all variables. The participants were 165 children with AD/HD who were part of the Multimodal Treatment Study of Children With ADHD and their 1,298 classmates. RESULTS: Variables that best discriminated between children with ADHD and their classmates included peer rejection and negative imbalance between given and received liking ratings (i.e., children with ADHD liked others more than they were liked). CONCLUSION: Peer rejection and negative imbalance show most promise for identifying clinically significant levels of peer relationship impairment in children with ADHD. (J. of Att. Dis. 2009; 12(4) 372 380). PMID- 18388252 TI - On positive psychological outcomes: what helps groups with a history of conflict to forgive and reconcile with each other? AB - Three studies examined the roles of traditional and novel social psychological variables involved in intergroup forgiveness. Study 1 (N = 480) revealed that among the pro-Pinochet and the anti-Pinochet groups in Chile, forgiveness was predicted by ingroup identity (negatively), common ingroup identity (positively), empathy and trust (positively), and competitive victimhood (the subjective sense of having suffered more than the outgroup, negatively). Political ideology (Right vs. Left) moderated the relationship between empathy and forgiveness, trust and forgiveness, and between the latter and competitive victimhood. Study 2 (N = 309), set in the Northern Irish conflict between Protestants and Catholics, provided a replication and extension of Study 1. Finally, Study 3 (N = 155/108) examined the longitudinal relationship between forgiveness and reconciliation in Northern Ireland, revealing that forgiveness predicted reconciliation intentions. The reverse direction of this relationship was also marginally significant. Results are discussed in terms of their theoretical and practical implications. PMID- 18388253 TI - Neurophysiological measures of involuntary and voluntary attention allocation and dispositional differences in need for cognition. AB - Need for cognition (NFC) refers to stable individual differences in the intrinsic motivation to engage in and enjoy effortful cognitive endeavors and has been a useful predictor of dispositional differences in information processing. Although cognitive resource allocation conceptualized as cognitive effort is assumed to be the key mediator of NFC-specific processing, to date no research has systematically addressed its underpinnings. Using a neurocognitive paradigm and recording event-related potentials associated with bottom-up and top-down-driven aspects of attention, the present research contributes to filling this gap. In Study 1, high-NFC individuals showed larger P3a amplitudes to contextually novel events, indicating greater involuntary (automatic) attention allocation. This effect was replicated in Study 2, where NFC also was positively correlated with the P3b to target stimuli, indicating voluntary (controlled) processes of attention allocation. Thus, our findings provide first evidence for neurophysiological correlates of NFC and can improve the understanding of NFC specific processing. PMID- 18388254 TI - Determinants of task performance and invested effort: a need for closure by relative cognitive capacity interaction analysis. AB - This study examines the simultaneous effects of need for closure (NFC) and relative cognitive capacity on invested effort and task performance within the integrative analysis framework using behavioral data. Two main results were obtained. First, the authors revealed a significant interaction effect between relative cognitive capacity (manipulated through task difficulty) and NFC (manipulated through time pressure, noise, and fear of invalidity as well as assessed by an individual differences measure) on effort investment. Second, contrary to dispositional NFC, manipulations yielded a "dual effect" because they negatively affected task performance as well as invested effort. The latter result was interpreted as an indication that noise and time pressure manipulations also tax cognitive resources. The two main findings are discussed and the authors go further into the divergences between dispositional and manipulated NFC. PMID- 18388255 TI - On the immediate consequences of intergroup categorization: activation of approach and avoidance motor behavior toward ingroup and outgroup members. AB - The present research argues that intergroup categorization has immediate behavioral consequences. Specifically, intergroup categorization is hypothesized to prepare the organism to respond differently to ingroup and outgroup members so that approach-like motor movements should be faster toward ingroup- versus outgroup-related stimuli. In contrast, avoidance-like behavior should be facilitated when reacting to outgroup versus ingroup members. Studies 1 and 2 test the basic hypothesis in relation to ethnic, national, age, and political categorization. Study 3 uses a minimal group paradigm to test the hypothesis in relation to newly formed groups. Across these experiments, participants were generally faster in performing approach-like motor movements toward ingroup members or avoidance behaviors toward outgroup members. The evolutionary function and the cognitive underpinnings of this state of "physical readiness" to approach ingroup and avoid outgroup members are discussed. PMID- 18388256 TI - Patients' perception of the informed consent process for neurooncology clinical trials. AB - The informed consent process is a cornerstone of modern medical research. This study was conducted to explore the process in the context of neurooncology clinical trials. Qualitative methodology and analysis were used on open-ended, face-to-face interviews conducted with 21 patients. Six comprehensive themes emerged: (1) general understanding of the objectives and purpose of clinical trials was good, (2) recall of risks was low, (3) patients did not believe that their care would be compromised by forgoing the clinical trial, (4) patients felt participation was voluntary and free of coercion, (5) patients would not have withdrawn from the trial in the event of complications, and (6) patients were satisfied with the informed consent process. Informed consent is a dynamic process; when appropriately executed, it can be a powerful safeguard protecting patient autonomy. If sufficient time is allowed to deliberate participation and ample opportunity is provided for information sharing and disclosure, researchers can be confident that participants are knowledgeable about the trial and aware of their rights. PMID- 18388257 TI - Simvastatin inhibits Staphylococcus aureus host cell invasion through modulation of isoprenoid intermediates. AB - Patients on a statin regimen have a decreased risk of death due to bacterial sepsis. We have found that protection by simvastatin includes the inhibition of host cell invasion by Staphylococcus aureus, the most common etiologic agent of sepsis. Inhibition was due in part to depletion of isoprenoid intermediates within the cholesterol biosynthesis pathway and led to the cytosolic accumulation of the small GTPases CDC42, Rac, and RhoB. Actin stress fiber disassembly required for host invasion was attenuated by simvastatin and by the inhibition of phosphoinositide 3-kinase (PI3K) activity. PI3K relies on coupling to prenylated proteins, such as this subset of small GTPases, for access to membrane-bound phosphoinositide to mediate stress fiber disassembly. Therefore, we examined whether simvastatin restricts PI3K cellular localization. In response to simvastatin, the PI3K isoform p85, coupled to these small-GTPases, was sequestered within the cytosol. From these findings, we propose a mechanism whereby simvastatin restricts p85 localization, inhibiting the actin dynamics required for bacterial endocytosis. This approach may provide the basis for protection at the level of the host in invasive infections by S. aureus. PMID- 18388258 TI - Spiro[imidazo[1,2-a]pyridine-3,2-indan]-2(3H)-one (ZSET1446/ST101) treatment rescues olfactory bulbectomy-induced memory impairment by activating Ca2+/calmodulin kinase II and protein kinase C in mouse hippocampus. AB - Olfactory bulbectomy (OBX) in mice elicits impaired memory and cognitive functions. Here, we found that chronic oral administration of spiro[imidazo[1,2 a]pyridine-3,2-indan]-2(3H)-one (ZSET1446/ST101) (0.1-1 mg/kg/day), a novel cognitive enhancer, significantly improved memory deficits as assessed by Y-maze and novel object recognition tasks in OBX mice. Immunostaining of cholinergic neurons in the medial septum by using an anti-choline acetyltransferase antibody indicated that chronic ZSET1446 treatment did not rescue cholinergic neurons. However, chronic treatment significantly restored OBX-induced decreases both in calcium/calmodulin-dependent protein kinase II (CaMKII) and protein kinase C (PKC) phosphorylation without improving decreased extracellular signal-regulated kinase phosphorylation in the hippocampal CA1 region. Consistent with enhanced CaMKII and PKC phosphorylation, ZSET1446 treatment improved glutamate receptor 1 (Ser-831) phosphorylation in the hippocampal CA1 region. ZSET1446 treatment also significantly rescued impaired long-term potentiation (LTP) in the hippocampal CA1 region of OBX mice. Taken together, the cognition-enhancing effect of ZSET1446 is probably mediated in part by stimulation of CaMKII and PKC activities, which in turn rescue impaired hippocampal LTP in OBX mice. PMID- 18388259 TI - Fine structure constant defines visual transparency of graphene. AB - There are few phenomena in condensed matter physics that are defined only by the fundamental constants and do not depend on material parameters. Examples are the resistivity quantum, h/e2 (h is Planck's constant and e the electron charge), that appears in a variety of transport experiments and the magnetic flux quantum, h/e, playing an important role in the physics of superconductivity. By and large, sophisticated facilities and special measurement conditions are required to observe any of these phenomena. We show that the opacity of suspended graphene is defined solely by the fine structure constant, a = e2/hc feminine 1/137 (where c is the speed of light), the parameter that describes coupling between light and relativistic electrons and that is traditionally associated with quantum electrodynamics rather than materials science. Despite being only one atom thick, graphene is found to absorb a significant (pa = 2.3%) fraction of incident white light, a consequence of graphene's unique electronic structure. PMID- 18388260 TI - ROS-generating mitochondrial DNA mutations can regulate tumor cell metastasis. AB - Mutations in mitochondrial DNA (mtDNA) occur at high frequency in human tumors, but whether these mutations alter tumor cell behavior has been unclear. We used cytoplasmic hybrid (cybrid) technology to replace the endogenous mtDNA in a mouse tumor cell line that was poorly metastatic with mtDNA from a cell line that was highly metastatic, and vice versa. Using assays of metastasis in mice, we found that the recipient tumor cells acquired the metastatic potential of the transferred mtDNA. The mtDNA conferring high metastatic potential contained G13997A and 13885insC mutations in the gene encoding NADH (reduced form of nicotinamide adenine dinucleotide) dehydrogenase subunit 6 (ND6). These mutations produced a deficiency in respiratory complex I activity and were associated with overproduction of reactive oxygen species (ROS). Pretreatment of the highly metastatic tumor cells with ROS scavengers suppressed their metastatic potential in mice. These results indicate that mtDNA mutations can contribute to tumor progression by enhancing the metastatic potential of tumor cells. PMID- 18388262 TI - The promise of cancer research. PMID- 18388261 TI - DNA from pre-Clovis human coprolites in Oregon, North America. AB - The timing of the first human migration into the Americas and its relation to the appearance of the Clovis technological complex in North America at about 11,000 to 10,800 radiocarbon years before the present (14C years B.P.) remains contentious. We establish that humans were present at Paisley 5 Mile Point Caves, in south-central Oregon, by 12,300 14C years B.P., through the recovery of human mitochondrial DNA (mtDNA) from coprolites, directly dated by accelerator mass spectrometry. The mtDNA corresponds to Native American founding haplogroups A2 and B2. The dates of the coprolites are >1000 14C years earlier than currently accepted dates for the Clovis complex. PMID- 18388263 TI - AIDS research. Review of vaccine failure prompts a return to basics. PMID- 18388264 TI - Space science. NASA's Stern quits over Mars exploration plans. PMID- 18388265 TI - Microbiology. Germs take a bite out of antibiotics. PMID- 18388266 TI - Astronomy. China's LAMOST Observatory prepares for the ultimate test. PMID- 18388267 TI - Defense research. U.S. asked to bolster ties with China. PMID- 18388268 TI - Archaeology: DNA from fossil feces breaks Clovis barrier. PMID- 18388269 TI - Cancer biology. All in the stroma: cancer's Cosa Nostra. PMID- 18388270 TI - American Physical Society meeting. Magnetic measurements hint at toastier superconductivity. PMID- 18388271 TI - American Physical Society meeting. Laser plays chemical matchmaker. PMID- 18388272 TI - American Physical Society meeting. Squeeze play makes solid helium flow. PMID- 18388273 TI - Conservation biology. Puzzling over a Steller whodunit. PMID- 18388274 TI - Conserving top predators in ecosystems. PMID- 18388275 TI - The role of fisheries-induced evolution. PMID- 18388276 TI - Tips for NIH. PMID- 18388277 TI - Public health. A case study of personalized medicine. PMID- 18388278 TI - Biochemistry. Toward understanding self-splicing. PMID- 18388279 TI - Climate. Blooms like it hot. PMID- 18388280 TI - Development. Deconstructing pluripotency. PMID- 18388281 TI - Cell signaling. Tel2 finally tells one story. PMID- 18388282 TI - Astronomy. Small-scale observations tell a cosmological story. PMID- 18388283 TI - Music theory. Creating musical variation. PMID- 18388284 TI - Stochasticity and cell fate. AB - Fundamental to living cells is the capacity to differentiate into subtypes with specialized attributes. Understanding the way cells acquire their fates is a major challenge in developmental biology. How cells adopt a particular fate is usually thought of as being deterministic, and in the large majority of cases it is. That is, cells acquire their fate by virtue of their lineage or their proximity to an inductive signal from another cell. In some cases, however, and in organisms ranging from bacteria to humans, cells choose one or another pathway of differentiation stochastically, without apparent regard to environment or history. Stochasticity has important mechanistic requirements. We speculate on why stochasticity is advantageous-and even critical in some circumstances-to the individual, the colony, or the species. PMID- 18388285 TI - Bats limit insects in a neotropical agroforestry system. AB - Exclosure experiments have demonstrated the effects of bird predation on arthropods. In a Mexican coffee plantation, we excluded foliage-gleaning bird and bat predators from coffee plants. Effects of bats and birds were additive. In the dry season, birds reduced arthropods in coffee plants by 30%; birds and bats together reduced arthropods by 46%. In the wet season, bats reduced arthropods by 84%, whereas birds reduced them by only 58%. We conclude that previous "bird" exclosure experiments may have systematically underestimated the effects of bats. PMID- 18388286 TI - Bats limit arthropods and herbivory in a tropical forest. AB - Previous exclosure studies measuring the top-down control of arthropod abundance and herbivory combined the effects of birds and bats. We experimentally partitioned bird predation from bat predation in a lowland tropical forest in Panama and measured the direct effects (arthropod abundance) and indirect effects (herbivory). The exclusion of birds and bats each directly increased arthropod abundance on plants: Bird-exclosed plants contained 65% more, and bat-exclosed plants 153% more, arthropods than controls. Birds and bats also indirectly increased herbivory: Bird-exclosed plants suffered 67% more, and bat-exclosed plants 209% more, herbivory than controls. We conclude that bats have dramatic ecological effects that were previously overlooked. PMID- 18388287 TI - Aztec arithmetic revisited: land-area algorithms and Acolhua congruence arithmetic. AB - Acolhua-Aztec land records depicting areas and side dimensions of agricultural fields provide insight into Aztec arithmetic. Hypothesizing that recorded areas resulted from indigenous calculation, in a study of sample quadrilateral fields we found that 60% of the area values could be reproduced exactly by computation. In remaining cases, discrepancies between computed and recorded areas were consistently small, suggesting use of an unknown indigenous arithmetic. In revisiting the research, we discovered evidence for the use of congruence principles, based on proportions between the standard linear Acolhua measure and their units of shorter length. This procedure substitutes for computation with fractions and is labeled "Acolhua congruence arithmetic." The findings also clarify variance between Acolhua and Tenochca linear units, long an issue in understanding Aztec metrology. PMID- 18388288 TI - Crystal structure of a self-spliced group II intron. AB - Group II introns are self-splicing ribozymes that catalyze their own excision from precursor transcripts and insertion into new genetic locations. Here we report the crystal structure of an intact, self-spliced group II intron from Oceanobacillus iheyensis at 3.1 angstrom resolution. An extensive network of tertiary interactions facilitates the ordered packing of intron subdomains around a ribozyme core that includes catalytic domain V. The bulge of domain V adopts an unusual helical structure that is located adjacent to a major groove triple helix (catalytic triplex). The bulge and catalytic triplex jointly coordinate two divalent metal ions in a configuration that is consistent with a two-metal ion mechanism for catalysis. Structural and functional analogies support the hypothesis that group II introns and the spliceosome share a common ancestor. PMID- 18388289 TI - Revealing magnetic interactions from single-atom magnetization curves. AB - The miniaturization of magnetic devices toward the limit of single atoms calls for appropriate tools to study their magnetic properties. We demonstrate the ability to measure magnetization curves of individual magnetic atoms adsorbed on a nonmagnetic metallic substrate with use of a scanning tunneling microscope with a spin-polarized tip. We can map out low-energy magnetic interactions on the atomic scale as evidenced by the oscillating indirect exchange between a Co adatom and a nanowire on Pt(111). These results are important for the understanding of variations that are found in the magnetic properties of apparently identical adatoms because of different local environments. PMID- 18388290 TI - The roles of subsurface carbon and hydrogen in palladium-catalyzed alkyne hydrogenation. AB - Alkynes can be selectively hydrogenated into alkenes on solid palladium catalysts. This process requires a strong modification of the near-surface region of palladium, in which carbon (from fragmented feed molecules) occupies interstitial lattice sites. In situ x-ray photoelectron spectroscopic measurements under reaction conditions indicated that much less carbon was dissolved in palladium during unselective, total hydrogenation. Additional studies of hydrogen content using in situ prompt gamma activation analysis, which allowed us to follow the hydrogen content of palladium during catalysis, indicated that unselective hydrogenation proceeds on hydrogen-saturated beta hydride, whereas selective hydrogenation was only possible after decoupling bulk properties from the surface events. Thus, the population of subsurface sites of palladium, by either hydrogen or carbon, governs the hydrogenation events on the surface. PMID- 18388291 TI - The electrical conductivity of post-perovskite in Earth's D'' layer. AB - Recent discovery of a phase transition from perovskite to post-perovskite suggests that the physical properties of Earth's lowermost mantle, called the D'' layer, may be different from those of the overlying mantle. We report that the electrical conductivity of (Mg0.9Fe0.1)SiO3 post-perovskite is >10(2) siemens per meter and does not vary greatly with temperature at the conditions of the D'' layer. A post-perovskite layer above the core-mantle boundary would, by electromagnetic coupling, enhance the exchange of angular momentum between the fluid core and the solid mantle, which can explain the observed changes in the length of a day on decadal time scales. Heterogeneity in the conductivity of the lowermost mantle is likely to depend on changes in chemistry of the boundary region, not fluctuations in temperature. PMID- 18388292 TI - Bacteria subsisting on antibiotics. AB - Antibiotics are a crucial line of defense against bacterial infections. Nevertheless, several antibiotics are natural products of microorganisms that have as yet poorly appreciated ecological roles in the wider environment. We isolated hundreds of soil bacteria with the capacity to grow on antibiotics as a sole carbon source. Of 18 antibiotics tested, representing eight major classes of natural and synthetic origin, 13 to 17 supported the growth of clonal bacteria from each of 11 diverse soils. Bacteria subsisting on antibiotics are surprisingly phylogenetically diverse, and many are closely related to human pathogens. Furthermore, each antibiotic-consuming isolate was resistant to multiple antibiotics at clinically relevant concentrations. This phenomenon suggests that this unappreciated reservoir of antibiotic-resistance determinants can contribute to the increasing levels of multiple antibiotic resistance in pathogenic bacteria. PMID- 18388293 TI - Reversible compartmentalization of de novo purine biosynthetic complexes in living cells. AB - Purines are synthesized de novo in 10 chemical steps that are catalyzed by six enzymes in eukaryotes. Studies in vitro have provided little evidence of anticipated protein-protein interactions that would enable substrate channeling and regulation of the metabolic flux. We applied fluorescence microscopy to HeLa cells and discovered that all six enzymes colocalize to form clusters in the cellular cytoplasm. The association and dissociation of these enzyme clusters can be regulated dynamically, by either changing the purine levels of or adding exogenous agents to the culture media. Collectively, the data provide strong evidence for the formation of a multi-enzyme complex, the "purinosome," to carry out de novo purine biosynthesis in cells. PMID- 18388295 TI - Entrainment of neuronal oscillations as a mechanism of attentional selection. AB - Whereas gamma-band neuronal oscillations clearly appear integral to visual attention, the role of lower-frequency oscillations is still being debated. Mounting evidence indicates that a key functional property of these oscillations is the rhythmic shifting of excitability in local neuronal ensembles. Here, we show that when attended stimuli are in a rhythmic stream, delta-band oscillations in the primary visual cortex entrain to the rhythm of the stream, resulting in increased response gain for task-relevant events and decreased reaction times. Because of hierarchical cross-frequency coupling, delta phase also determines momentary power in higher-frequency activity. These instrumental functions of low frequency oscillations support a conceptual framework that integrates numerous earlier findings. PMID- 18388294 TI - Single-molecule DNA sequencing of a viral genome. AB - The full promise of human genomics will be realized only when the genomes of thousands of individuals can be sequenced for comparative analysis. A reference sequence enables the use of short read length. We report an amplification-free method for determining the nucleotide sequence of more than 280,000 individual DNA molecules simultaneously. A DNA polymerase adds labeled nucleotides to surface-immobilized primer-template duplexes in stepwise fashion, and the asynchronous growth of individual DNA molecules was monitored by fluorescence imaging. Read lengths of >25 bases and equivalent phred software program quality scores approaching 30 were achieved. We used this method to sequence the M13 virus to an average depth of >150x and with 100% coverage; thus, we resequenced the M13 genome with high-sensitivity mutation detection. This demonstrates a strategy for high-throughput low-cost resequencing. PMID- 18388296 TI - Episodic-like memory in rats: is it based on when or how long ago? AB - Recent experiments with rats suggest that they show episodic-like or what-where when memory for a preferred food found on a radial maze. Although memory for when a salient event occurred suggests that rats can mentally travel in time to a moment in the past, an alternative possibility is that they remember how long ago the food was found. Three groups of rats were tested for memory of previously encountered food. The different groups could use only the cues of when, how long ago, or when + how long ago. Only the cue of how long ago food was encountered was used successfully. These results suggest that episodic-like memory in rats is qualitatively different from human episodic memory. PMID- 18388298 TI - Antiviral NK cell responses in HIV infection: I. NK cell receptor genes as determinants of HIV resistance and progression to AIDS. AB - NK cells play an important role in controlling viral infections. They can kill virus-infected cells directly as well as indirectly via antibody-dependent, cell mediated cytotoxicity. They need no prior sensitization and expansion for this killing. NK cells are also considered as important regulators of antiviral immune responses. They do so by secreting a multitude of soluble mediators and by directly interacting with other immune cells, e.g., dendritic cells. NK cells do not possess a single well-defined receptor to recognize antigens on target cells. Instead, they express an array of inhibitory and activating receptors and coreceptors, which bind to their cognate ligands expressed on the surface of target cells. These ligands include classical and nonclassical MHC class I antigens, MHC-like proteins, and a variety of other self- and virus-derived molecules. They may be expressed constitutively and/or de novo on the surface of virus-infected cells. NK cell receptors (NKRs) of the killer-cell Ig-like receptor (KIR) family, like their MHC class I ligands, are highly polymorphic. Several recent studies suggest that epistatic interactions between certain KIR and MHC class I genes may determine innate resistance of the host to viral infections, including HIV. In the first part of this review article, we provide an overview of the current state of knowledge of NK cell immunobiology and describe how NKR genes, alone and in combination with HLA genes, may determine genetic resistance/susceptibilty to HIV infection and the development of AIDS in humans. PMID- 18388297 TI - CD44 mobilization in allogeneic dendritic cell-T cell immunological synapse plays a key role in T cell activation. AB - CD44 is involved in several biological processes owing to its dual role as a cell adhesion and signaling molecule. In an allogeneic dendritic cell (DC)-T cell interaction model, we show here that CD44 gets clustered at the contact between T cells with mature but not immature DCs. Also, CD44 colocalized with lipid rafts at the immunological synapse (IS). Using DCs or T cells derived from CD44 deficient mice, we observed that the presence of CD44 on DCs and T cells is important for the formation of DC-T cell tight conjugates. However, deficiency of CD44 on DCs but not T cells affected the functional IS, as indicated by decreased phosphotyrosine and protein kinase C-theta enrichment at the synapse. Also, CD44 deficient DCs induced significantly decreased proliferation as well as IL-2 and IFN-gamma production from allogeneic T cells. The polarization of CD44 at the synapse was also noted in an antigen (OVA)-specific, syngeneic DC-T cell interaction using OVA-specific T cells derived from OT-II mice. It was believed that large molecules such as CD44 were excluded from the IS. Results presented here show for the first time that CD44 is recruited to the IS during allogeneic DC and T cell interactions and plays an important role in subsequent T cell activation. PMID- 18388299 TI - Antiviral NK cell responses in HIV infection: II. viral strategies for evasion and lessons for immunotherapy and vaccination. AB - As is the case in other viral infections, humans respond to HIV infection by activating their NK cells. However, the virus uses several strategies to neutralize and evade the host's NK cell responses. Consequently, it is not surprising that NK cell functions become compromised in HIV-infected individuals in early stages of the infection. The compromised NK cell functions also adversely affect several aspects of the host's antiviral adaptive immune responses. Researchers have made significant progress in understanding how HIV counters NK cell responses of the host. This knowledge has opened new avenues for immunotherapy and vaccination against this infection. In the first part of this review article, we gave an overview of our current knowledge of NK cell biology and discussed how the genes encoding NK cell receptors and their ligands determine innate genetic resistance/susceptibilty of humans against HIV infections and AIDS. In this second part, we discuss NK cell responses, viral strategies to counter these responses, and finally, their implications for anti HIV immunotherapy and vaccination. PMID- 18388300 TI - Diminishment of alpha-MSH anti-inflammatory activity in MC1r siRNA-transfected RAW264.7 macrophages. AB - The neuropeptide alpha-melanocyte-stimulating hormone (alpha-MSH) is a powerful suppressor of inflammation mediated by macrophages, which express at least two receptors, melanocortin 1 and 3 receptors (MC1r and MC3r) that bind alpha-MSH. Albeit, the anti-inflammatory activity of alpha-MSH has been well documented in macrophages, the mechanisms of alpha-MSH activity in macrophages are not clearly understood. This study is to investigate which of the MCr expressed on macrophages is associated with the immunosuppressive activities of alpha-MSH on LPS-stimulated macrophages. To address this question, we transfected RAW264.7 macrophage cells with MC1r small interfering (si)RNA, which specifically targets mouse MC1r mRNA. The diminution of MC1r mRNA expression was 82% at 24 h and 67% at 48 h after transfection. There was a significant loss in alpha-MSH suppression of NO generation and TNF-alpha production by MC1r siRNA-transfected macrophages stimulated with LPS. There was an equally diminished alpha-MSH suppression of LPS stimulated intracellular activation of NF-kappaB and p38 phosphorylation. In addition, the diminishment of MC1r expression by siRNA transfection had no influence on MC3r expression and function in the macrophages. These findings demonstrate that alpha-MSH suppression of LPS-induced inflammatory activity in macrophages requires expression of MC1r. The results imply that although all of the MCr are G-coupled proteins, they may not necessarily function through the same intracellular pathways in macrophages. PMID- 18388301 TI - Human bone marrow adipocytes block granulopoiesis through neuropilin-1-induced granulocyte colony-stimulating factor inhibition. AB - Adipocytes are part of hematopoietic microenvironment, even though up to now in humans, their role in hematopoiesis is still questioned. We have previously shown that accumulation of fat cells in femoral bone marrow (BM) coincides with increased expression of neuropilin-1 (NP-1), while it is weakly expressed in hematopoietic iliac crest BM. Starting from this observation, we postulated that adipocytes might exert a negative effect on hematopoiesis mediated through NP-1. To test this hypothesis, we set up BM adipocytes differentiated into fibroblast like fat cells (FLFC), which share the major characteristics of primitive unilocular fat cells, as an experimental model. As expected, FLFCs constitutively produced macrophage colony stimulating factor and induced CD34(+) differentiation into macrophages independently of cell-to-cell contact. By contrast, granulopoiesis was hampered by cell-to-cell contact but could be restored in transwell culture conditions, together with granulocyte colony stimulating factor production. Both functions were also recovered when FLFCs cultured in contact with CD34(+) cells were treated with an antibody neutralizing NP-1, which proved its critical implication in contact inhibition. An inflammatory cytokine such as interleukin-1 beta or dexamethasone modulates FLFC properties to restore granulopoiesis. Our data provide the first evidence that primary adipocytes exert regulatory functions during hematopoiesis that might be implicated in some pathological processes. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388302 TI - Neuropeptide Y promotes neurogenesis in murine subventricular zone. AB - Stem cells of the subventricular zone (SVZ) represent a reliable source of neurons for cell replacement. Neuropeptide Y (NPY) promotes neurogenesis in the hippocampal subgranular layer and the olfactory epithelium and may be useful for the stimulation of SVZ dynamic in brain repair purposes. We describe that NPY promotes SVZ neurogenesis. NPY (1 microM) treatments increased proliferation at 48 hours and neuronal differentiation at 7 days in SVZ cell cultures. NPY proneurogenic properties are mediated via the Y1 receptor. Accordingly, Y1 receptor is a major active NPY receptor in the mouse SVZ, as shown by functional autoradiography. Moreover, short exposure to NPY increased immunoreactivity for the phosphorylated form of extracellular signal-regulated kinase 1/2 in the nucleus, compatible with a trigger for proliferation, whereas 6 hours of treatment amplified the phosphorylated form of c-Jun-NH(2)-terminal kinase signal in growing axons, consistent with axonogenesis. NPY, as a promoter of SVZ neurogenesis, is a crucial factor for future development of cell-based brain therapy. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388303 TI - Assessment of stromal-derived inducing activity in the generation of dopaminergic neurons from human embryonic stem cells. AB - Producing dopaminergic (DA) neurons is a major goal of human embryonic stem cell (hESC) research. DA neurons can be differentiated from hESC by coculture with the mouse PA6 stromal cell line; this differentiation-inducing effect is termed stromal-derived inducing activity (SDIA). The molecular and biochemical nature of SDIA is, however, unknown. Various studies have suggested that SDIA involves either a fixation-resistant component located on the PA6 cell surface or factors secreted into the medium by PA6 cells. To address this question, hESC were cocultured with PA6 cells for 12 days and then further differentiated with sonic hedgehog homolog, fibroblast growth factor-8, and glial cell line-derived neurotrophic factor. After 18 days, 34% of cells were tyrosine hydroxylase (TH)+. When PA6 cells were fixed or irradiated, the number of TH+ cells was decreased by threefold, whereas mitomycin-c treatment of feeder cells decreased the number of TH+ cells by 32%. The neural-inducing effect of PA6 cells, as monitored by beta III-tubulin expression, was minimally affected by mitomycin-c treatment or fixation but was decreased 50% by irradiation. Medium conditioned by PA6 cells was ineffective in differentiating TH+ cells when used alone. Conditioned medium combined with heparin and/or fixed PA6 cells produced TH+ cell differentiation, although less effectively than PA6 cell coculture. Thus, PA6 cell surface activity is required for neural differentiation of hESC, but secreted factors are required for the specific DA neuron-inducing effect. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388304 TI - The neuronal differentiation potential of Ldb1-null mutant embryonic stem cells is dependent on extrinsic influences. AB - LIM-domain binding protein 1 (Ldb1) is a multiadaptor protein that mediates the action of transcription factors, including LIM-homeodomain proteins. To elucidate the functional role of Ldb1 in the neuronal differentiation of embryonic stem (ES) cells, we have generated Ldb1-null mutant (Ldb1-/-) ES cells and examined neuronal differentiation potentials in vitro using two different neuronal differentiation protocols. When subjected to a five-stage protocol that recapitulates in vivo conditions of neuronal differentiation, wild-type ES cells differentiated into a wide spectrum of neuronal cell types. However, Ldb1-/- ES cells did not differentiate into neuronal cells; instead, they differentiated into sarcomeric alpha-actinin-positive muscle cells. In contrast, when an adherent monolayer culture procedure (which is based on the default mechanism of neural induction and eliminates environmental influences) was applied, both wild type and Ldb1-/- ES cells differentiated into MAP2-positive mature neurons. Comparison of the results obtained when two different neuronal differentiation protocols were used suggests that Ldb1-/- ES cells have an innate potential to differentiate into neuronal cells, but this potential can be inhibited by environmental influences. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388305 TI - Concise review: adult multipotent stromal cells and cancer: risk or benefit? AB - This review focuses on the interaction between multipotent stromal cells (MSCs) and carcinoma and the possible use of MSCs in cell-based anticancer therapies. MSCs are present in multiple tissues and are defined as cells displaying the ability to differentiate in multiple lineages, including chondrocytes, osteoblasts, and adipocytes. Recent evidence also suggests that they could play a role in the progression of carcinogenesis and that MSCs could migrate toward primary tumors and metastatic sites. It is possible that MSCs could also be involved in the early stages of carcinogenesis through spontaneous transformation. In addition, it is thought that MSCs can modulate tumor growth and metastasis, although this issue remains controversial and not well understood. The immunosuppressive properties and proangiogenic properties of MSCs account, at least in part, for their effects on cancer development. On the other hand, cancer cells also have the ability to enhance MSC migration. This complex dialog between MSCs and cancer cells is certainly critical for the outcome of tumor development. Interestingly, several studies have shown that MSCs engineered to express antitumor factors could be an innovative choice as a cell-mediated gene therapy to counteract tumor growth. More evidence will be needed to understand how MSCs positively or negatively modulate carcinogenesis and to evaluate the safety of MSC use in cell-mediated gene strategies. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388306 TI - Regulation of self-renewal and pluripotency by Sox2 in human embryonic stem cells. AB - Human embryonic stem (hES) cells, derived from blastocysts, are capable of unlimited self-renewal and differentiation into all cell lineages of the body. Because of their pluripotent nature, hES cells are valuable tools for understanding human development and advancing the field of regenerative medicine. However, one key to harnessing the therapeutic power of hES cells for biomedical applications begins with determining how these cells maintain their pluripotent and undifferentiated state. Studies in mice have implicated three factors in regulating pluripotency in embryonic stem cells, Oct4, Nanog, and Sox2. However, significant differences in growth regulation between mouse embryonic stem and hES cells have been identified, suggesting a need to determine when and how factors work in hES cells. To date, the transcription factors Oct4 and Nanog have been identified as critical regulators of stem cell fate by functional studies in hES cells. To determine the role of Sox2 in maintaining hES cell pluripotency and self-renewal, we used RNA interference to specifically knock down Sox2 gene expression. Reduction of Sox2 expression in hES cells results in loss of the undifferentiated stem cell state, as indicated by a change in cell morphology, altered stem cell marker expression, and increased expression of trophectoderm markers. In addition, knockdown of Sox2 results in reduced expression of several key stem cell factors, including Oct4 and Nanog, linking these three factors together in a pluripotent regulatory network. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388308 TI - Local delivery of granulocyte colony stimulating factor-mobilized CD34-positive progenitor cells using bioscaffold for modality of unhealing bone fracture. AB - We recently reported that i.v. transplantation of adult human circulating CD34+ cells, an endothelial/hematopoietic progenitor-enriched cell population, contributes to fracture healing through the enhancement of vasculogenesis and osteogenesis. However, the scarcity of CD34+ cells in the adult human is a critical issue for the future clinical application of this method. To overcome this issue, we assessed in vitro and in vivo capacity of granulocyte colony stimulating factor-mobilized peripheral blood (GM-PB) human CD34+ cells for vasculogenesis and osteogenesis. First, we confirmed the differentiation capability of GM-PB CD34+ cells into osteoblasts in vitro. Second, local transplantation of GM-PB CD34+ cells on atelocollagen scaffold was performed in nude rats in a model of unhealing fractures. Immunostaining for human leukocyte antigen-ABC of tissue samples 1 week after fracture and cell therapy showed the superior incorporation after local transplantation compared with systemic infusion. Third, the effects of local transplantation of 10(5) (Hi), 10(4) (Mid), or 10(3) (Lo) doses of GM-PB CD34+ cells or phosphate-buffered saline (PBS) on fracture healing were compared. Extrinsic vasculogenic and osteogenic differentiation of GM-PB CD34+ cells, enhancement of the intrinsic angio osteogenesis by recipient cells, augmentation of blood flow recovery at the fracture sites, and radiological and histological confirmation of fracture healing were observed only in the Hi and Mid groups but not in the Lo and PBS groups. These results strongly suggest that local transplantation of GM-PB CD34+ cells with atelocollagen scaffold is a feasible strategy for therapeutic vasculogenesis and osteogenesis needed for fracture healing. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388307 TI - Embryonic stem cell-derived Pitx3-enhanced green fluorescent protein midbrain dopamine neurons survive enrichment by fluorescence-activated cell sorting and function in an animal model of Parkinson's disease. AB - Both fetal ventral mesencephalic (VM) and embryonic stem (ES) cell-derived dopamine neurons have been used successfully to correct behavioral responses in animal models of Parkinson's disease. However, grafts derived from fetal VM cells or from ES cells contain multiple cell types, and the majority of these cells are not dopamine neurons. Isolation of ES cell-derived dopamine neurons and subsequent transplantation would both elucidate the capacity of these neurons to provide functional input and also further explore an efficient and safer use of ES cells for the treatment of Parkinson's disease. Toward this goal, we used a Pitx3-enhanced green fluorescent protein (Pitx3-eGFP) knock-in mouse blastocyst derived embryonic stem (mES) cell line and fluorescence-activated cell sorting (FACS) to select and purify midbrain dopamine neurons. Initially, the dopaminergic marker profile of intact Pitx3-eGFP mES cultures was evaluated after differentiation in vitro. eGFP expression overlapped closely with that of Pitx3, Nurr1, Engrailed-1, Lmx1a, tyrosine hydroxylase (TH), l-aromatic amino acid decarboxylase (AADC), and vesicular monoamine transporter 2 (VMAT2), demonstrating that these cells were of a midbrain dopamine neuron character. Furthermore, postmitotic Pitx3-eGFP(+) dopamine neurons, which constituted 2%-5% of all live cells in the culture after dissociation, could be highly enriched to >90% purity by FACS, and these isolated neurons were viable, extended neurites, and maintained a dopaminergic profile in vitro. Transplantation to 6 hydroxydopamine-lesioned rats showed that an enriched dopaminergic population could survive and restore both amphetamine- and apomorphine-induced functions, and the grafts contained large numbers of midbrain dopamine neurons, which innervated the host striatum. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18388309 TI - Cell polarity and cancer--cell and tissue polarity as a non-canonical tumor suppressor. AB - Correct establishment and maintenance of cell polarity is required for the development and homeostasis of all metazoans. Cell-polarity mechanisms are responsible not only for the diversification of cell shapes but also for regulation of the asymmetric cell divisions of stem cells that are crucial for their correct self-renewal and differentiation. Disruption of cell polarity is a hallmark of cancer. Furthermore, recent evidence indicates that loss of cell polarity is intimately involved in cancer: several crucial cell-polarity proteins are known proto-oncogenes or tumor suppressors, basic mechanisms of cell polarity are often targeted by oncogenic signaling pathways, and deregulation of asymmetric cell divisions of stem or progenitor cells may be responsible for abnormal self-renewal and differentiation of cancer stem cells. Data from in vivo and three-dimensional (3D) cell-culture models demonstrate that tissue organization attenuates the phenotypic outcome of oncogenic signaling. We suggest that polarized 3D tissue organization uses cell-cell and cell-substratum adhesion structures to reinforce and maintain the cell polarity of pre-cancerous cells. In this model, polarized 3D tissue organization functions as a non-canonical tumor suppressor that prevents the manifestation of neoplastic features in mutant cells and, ultimately, suppresses tumor development and progression. PMID- 18388310 TI - CCAAT/enhancer binding protein beta deficiency provides cerebral protection following excitotoxic injury. AB - The CCAAT/enhancer-binding protein beta (C/EBPbeta, also known as CEBPB) was first identified as a regulator of differentiation and inflammatory processes in adipose tissue and liver. Although C/EBPbeta was initially implicated in synaptic plasticity, its function in the brain remains largely unknown. We have previously shown that C/EBPbeta regulates the expression of genes involved in inflammatory processes and brain injury. Here, we have demonstrated that the expression of C/EBPbeta is notably increased in the hippocampus in a murine model of excitotoxicity. Mice lacking C/EBPbeta showed a reduced inflammatory response after kainic acid injection, and exhibited a dramatic reduction in pyramidal cell loss in the CA1 and CA3 subfields of the hippocampus. These data reveal an essential function for C/EBPbeta in the pathways leading to excitotoxicity mediated damage and suggest that inhibitors of this transcription factor should be evaluated as possible neuroprotective therapeutic agents. PMID- 18388311 TI - An alternatively spliced isoform of PECAM-1 is expressed at high levels in human and murine tissues, and suggests a novel role for the C-terminus of PECAM-1 in cytoprotective signaling. AB - The Ig-ITIM family member PECAM-1 is expressed in vascular and endothelial cells, and its functions include suppression of mitochondria-dependent apoptosis. Previous studies have identified distinct PECAM-1 cytoplasmic domain splice variants at the mRNA, but not protein, level. Several relatively abundant mRNA isoforms lack exon 15 (Delta15) and would theoretically encode a protein with a truncated cytoplasmic domain and a unique C-terminal sequence. Using a novel rabbit polyclonal antibody that specifically recognizes Delta15 PECAM-1, we found that the Delta15 PECAM-1 isoform was expressed in human tissues, including brain, testes and ovary. This isoform was also expressed on the cell surface of human platelets, human umbilical vein endothelial cells (HUVECs) and the Jurkat T-cell leukemia, human erythroleukemia (HEL) and U937 histiocytic lymphoma cell lines. Furthermore, murine platelets and lung lysates demonstrated abundant amounts of exon-15-deficient PECAM-1. Functional studies revealed that Delta15 PECAM-1 retains both its homophilic binding capacity and its ability to signal by means of its immunoreceptor tyrosine-based inhibitory motif (ITIM) domains. Delta15 PECAM-1 was unable, however, to protect against apoptosis induced by overexpression of Bax or treatment with the chemotherapy agent etoposide. These studies suggest a novel role for the PECAM-1 C-terminus in cytoprotective signaling and highlight a need for further characterization of expression of PECAM-1 isoforms in normal and malignant tissues. PMID- 18388313 TI - SNX9 regulates tubular invagination of the plasma membrane through interaction with actin cytoskeleton and dynamin 2. AB - Dynamic membrane remodeling during intracellular trafficking is controlled by the intricate interplay between lipids and proteins. BAR domains are modules that participate in endocytic processes by binding and deforming the lipid bilayer. Sorting nexin 9 (SNX9), which functions in clathrin-mediated endocytosis, contains a BAR domain, however, the properties of this domain are not well understood. Here we show that SNX9 shares many properties with other BAR domain containing proteins, such as amphiphysin and endophilin. SNX9 is able to deform the plasma membrane, as well as liposomes, into narrow tubules and recruit N-WASP and dynamin 2 to these tubules via its SH3 domain. SNX9-induced tubulation is antagonized by N-WASP and dynamin 2 while it is enhanced by perturbation of actin dynamics. However, SNX9 also has several unique properties. The tubulating activity requires the BAR and PX domains, as well as the low-complexity (LC) domain, which binds the Arp2/3 complex. SNX9 also binds to PtdIns(4)P-5-kinases via its PX domain and its tubulating activity is regulated by phosphoinositides. In addition, the kinase activity of PtdIns(4)P-5-kinases is stimulated by interaction with SNX9, suggesting a positive feedback interaction between SNX9 and PtdIns(4)P-5-kinases. These results suggest that SNX9 functions in the coordination of membrane remodeling and fission via interactions with actin regulating proteins, endocytic proteins and PtdIns(4,5)P2-metabolizing enzymes. PMID- 18388312 TI - Recycling of IRAP from the plasma membrane back to the insulin-responsive compartment requires the Q-SNARE syntaxin 6 but not the GGA clathrin adaptors. AB - Insulin recruits two transmembrane proteins, GLUT4 and IRAP, to the plasma membrane of muscle cells and adipocytes. The subcellular trafficking and localization of GLUT4, and to a lesser extent IRAP, have been intensely studied, yet the molecular mechanisms responsible for their insulin-responsive compartmentalization remain unknown. Herein we have investigated the endocytosis and recycling of IRAP from the cell surface back to the insulin-responsive compartment (IRC). Our results show that a key dileucine motif at position 76,77 (LL76,77), although required for the initial biosynthetic entry of IRAP into the IRC, is dispensable for entry into the IRC via the endosomal system. Indeed, we found that an AA76,77 mutant of IRAP is fully capable of undergoing endocytosis and is correctly routed back to the IRC. To verify that the AA76,77 mutant enters the bona fide IRC, we show that the internalized IRAP-AA76,77 construct is sequestered in an IRC that is insensitive to brefeldin A yet sensitive to a dominant-interfering mutant of AS160 (AS160-4P). In addition, we show that the GGA clathrin adaptors are not required for the re-entry of IRAP from the cell surface back into the IRC, whereas the Q-SNARE syntaxin 6 is required for this process. PMID- 18388314 TI - Internalization of LDL-receptor superfamily yolk-protein receptors during mosquito oogenesis involves transcriptional regulation of PTB-domain adaptors. AB - In the anautogenous disease vector mosquitoes Anopheles gambiae and Aedes aegypti, egg development is nutritionally controlled. A blood meal permits further maturation of developmentally repressed previtellogenic egg chambers. This entails massive storage of extraovarian yolk precursors by the oocyte, which occurs through a burst of clathrin-mediated endocytosis. Yolk precursors are concentrated at clathrin-coated structures on the oolemma by two endocytic receptors, the vitellogenin and lipophorin receptors. Both these mosquito receptors are members of the low-density-lipoprotein-receptor superfamily that contain FxNPxY-type internalization signals. In mammals, this tyrosine-based signal is not decoded by the endocytic AP-2 adaptor complex directly. Instead, two functionally redundant phosphotyrosine-binding domain adaptors, Disabled 2 and the autosomal recessive hypercholesterolemia protein (ARH) manage the internalization of the FxNPxY sorting signal. Here, we report that a mosquito ARH like protein, which we designate trephin, possess similar functional properties to the orthologous vertebrate proteins despite engaging AP-2 in an atypical manner, and that mRNA expression in the egg chamber is strongly upregulated shortly following a blood meal. Temporally regulated trephin transcription and translation suggests a mechanism for controlling yolk uptake when vitellogenin and lipophorin receptors are expressed and clathrin coats operate in previtellogenic ovaries. PMID- 18388315 TI - Influence of irofulven, a transcription-coupled repair-specific antitumor agent, on RNA polymerase activity, stability and dynamics in living mammalian cells. AB - Transcription-coupled repair (TCR) plays a key role in the repair of DNA lesions induced by bulky adducts and is initiated when the elongating RNA polymerase II (Pol II) stalls at DNA lesions. This is accompanied by alterations in Pol II activity and stability. We have previously shown that the monofunctional adducts formed by irofulven (6-hydroxymethylacylfulvene) are exclusively recognized by TCR, without involvement of global genome repair (GGR), making irofulven a unique tool to characterize TCR-associated processes in vivo. Here, we characterize the influence of irofulven on Pol II activity, stability and mobility in living mammalian cells. Our results demonstrate that irofulven induces specific inhibition of nucleoplasmic RNA synthesis, an important decrease of Pol II mobility, coupled to the accumulation of initiating polymerase and a time dependent loss of the engaged enzyme, associated with its polyubiquitylation. Both proteasome-mediated degradation of the stalled polymerase and new protein synthesis are necessary to allow Pol II recycling into preinitiating complexes. Together, our findings provide novel insights into the subsequent fate of the stalled RNA polymerase II and demonstrate the essential role of the recycling process for transcriptional reinitiation and viability of mammalian cells. PMID- 18388317 TI - Ypt1p is essential for retrograde Golgi-ER transport and for Golgi maintenance in S. cerevisiae. AB - The small GTPase Ypt1p of the Rab family is required for docking of ER-derived transport vesicles with the Golgi prior to fusion. However, the identity of the Rab protein that mediates docking of Golgi-derived COPI vesicles with the ER in retrograde transport remains elusive. Here, we show that in yeast Ypt1p is essential for retrograde transport from the Golgi to the ER. Retrieval of gpalphaF-HDEL (glycolylated pro-alpha-factor with an HDEL tag at the C-terminus) was blocked in Deltaypt1/SLY1-20 membranes at the restrictive temperature in vitro. Moreover, Ypt1p and the ER-resident t-SNARE Ufe1p interact genetically and biochemically, indicating a role for Ypt1p in consumption of COPI vesicles at the ER. Ypt1p is also essential for the maintenance of the morphology and the protein composition of the Golgi. Interestingly, the concentrations of the Golgi enzymes Anp1p and Mnn1p, the cargo protein Emp47p and the v-SNARE Sec22p were all substantially reduced in Golgi from a Deltaypt1/SLY1-20 strain as compared with wild-type Golgi, while the concentration of Arf1p and of coatomer were mildly affected. Finally, COPI vesicles generated from Deltaypt1/SLY1-20 Golgi membranes in vitro were depleted of Emp47p and Sec22p. These data demonstrate that Ypt1p plays an essential role in retrograde transport from the Golgi to the ER. PMID- 18388316 TI - Mob4 plays a role in spindle focusing in Drosophila S2 cells. AB - The characteristic bipolar shape of the mitotic spindle is produced by the focusing of the minus ends of microtubules at the spindle poles. The focus is maintained by the centrosome, a microtubule-nucleating organelle, as well as by proteins that are capable of focusing kinetochore fibers (K fibers) even in the absence of a centrosome. Here, we have performed a small-scale RNA interference (RNAi) screen of known or suspected pole-related proteins in Drosophila S2 cells. An unexpected outcome of this screen was the finding that one of the four Drosophila Mob proteins (a family of kinase regulators) plays a role in spindle pole organization. Time-lapse microscopy of mitotic cells depleted of Drosophila Mob4 by RNAi revealed that the K fibers splay apart and do not maintain their focus either in the presence or absence of functional centrosomes. The Mob4 RNAi phenotype most closely resembles that observed after depletion of the protein encoded by abnormal spindle (Asp), although Asp localization is not substantially affected by Mob4 RNAi. Expression of a Drosophila Mob4-GFP fusion protein revealed its localization to the nucleus in interphase and to spindle poles and kinetochores during mitosis. We propose that Mob4 in Drosophila controls a mitotic kinase that in turn regulates downstream target proteins involved in K fiber focusing at the poles. PMID- 18388318 TI - WASP and SCAR have distinct roles in activating the Arp2/3 complex during myoblast fusion. AB - Myoblast fusion takes place in two steps in mammals and in Drosophila. First, founder cells (FCs) and fusion-competent myoblasts (FCMs) fuse to form a trinucleated precursor, which then recruits further FCMs. This process depends on the formation of the fusion-restricted myogenic-adhesive structure (FuRMAS), which contains filamentous actin (F-actin) plugs at the sites of cell contact. Fusion relies on the HEM2 (NAP1) homolog Kette, as well as Blow and WASP, a member of the Wiskott-Aldrich-syndrome protein family. Here, we show the identification and characterization of schwachling--a new Arp3-null allele. Ultrastructural analyses demonstrate that Arp3 schwachling mutants can form a fusion pore, but fail to integrate the fusing FCM. Double-mutant experiments revealed that fusion is blocked completely in Arp3 and wasp double mutants, suggesting the involvement of a further F-actin regulator. Indeed, double-mutant analyses with scar/WAVE and with the WASP-interacting partner vrp1 (sltr, wip)/WIP show that the F-actin regulator scar also controls F-actin formation during myoblast fusion. Furthermore, the synergistic phenotype observed in Arp3 wasp and in scar vrp1 double mutants suggests that WASP and SCAR have distinct roles in controlling F-actin formation. From these findings we derived a new model for actin regulation during myoblast fusion. PMID- 18388319 TI - Actin-based propulsive forces and myosin-II-based contractile forces in migrating Dictyostelium cells. AB - It has been suggested that myosin II exerts traction forces at the posterior ends and retracting pseudopodia of migrating cells, but there is no direct evidence. Here, using a combination of total internal reflection fluorescence (TIRF) microscopy and force microscopy with a high spatial resolution of approximately 400 nm, we simultaneously recorded GFP-myosin II dynamics and traction forces under migrating Dictyostelium cells. Accumulation of filamentous myosin II and a subsequent increase in traction forces were detected in pseudopodia just before retraction. In the case of motorless myosin II, traction forces did not increase after accumulation, suggesting that the source of the retraction force is the motor activity of accumulated myosin II. Simultaneous recording of F-actin and traction forces revealed that traction forces were exerted under spot-like regions where F-actin accumulated. Cells migrated in a direction counter to the sum of the force vectors exerted at each spot, suggesting that the stress spots act as scaffolds to transmit the propulsive forces at the leading edge generated by actin polymerization. PMID- 18388320 TI - Dynamic regulation of ubiquitylation and deubiquitylation at the central spindle during cytokinesis. AB - During cytokinesis, the central spindle, a bundle of interdigitated anti-parallel microtubules between separating chromosomes, recruits various cytokinetic regulator proteins to the cleavage region. Here, we show that the level of protein ubiquitylation is strikingly and transiently elevated in Aurora B kinase positive double-band regions of the central spindle during cytokinesis. Two deubiquitylating enzymes UBPY and AMSH, which act on endosomes in interphase, were also recruited to the cleavage region. Whereas UBPY was detected only in the final stage of cytokinesis at the midbody, AMSH localized to a ring structure surrounding the mitotic kinesin MKLP1-positive region of the central spindle and midbody throughout cytokinesis. Depletion of cellular UBPY or AMSH led to defects in cytokinesis. VAMP8, a v-SNARE required for vesicle fusion in cytokinesis, localized to the central spindle region positive for ubiquitylated proteins, and underwent ubiquitylation and deubiquitylation by both UBPY and AMSH. Our results thus implicate the ubiquitylation/deubiquitylation of proteins including VAMP8 in cytokinesis. PMID- 18388321 TI - Centrosome separation driven by actin-microfilaments during mitosis is mediated by centrosome-associated tyrosine-phosphorylated cortactin. AB - The regulation of protein tyrosine phosphorylation is an important aspect during the cell cycle. From G2-M transition to mitotic anaphase, phosphorylation of Tyr421, Tyr466 and Tyr482 of cortactin, an actin-filament associated protein, is dramatically induced. The phosphorylated cortactin is almost exclusively associated with centrosomes or spindle poles during mitosis. At G2-M transition prior to the breakdown of the nuclear envelope, two duplicated centrosomes migrate towards opposite ends of the nucleus to form the spindle poles. This centrosome-separation process and also the start of mitosis are inhibited or delayed by the depolymerization of actin filaments. Also inhibited is the separation of centrosomes when a truncated form of cortactin is expressed, whose C-terminus contains the tyrosine phosphorylation region but lacks the actin binding domains. We introduced mutations at the tyrosine phosphorylation sites in the truncated C-terminus of cortactin and found that the C-terminus could no longer interfere with centrosome separation process. Our study shows that, cortactin phosphorylated at Tyr421, Tyr466 and Tyr482 mediates the actin-filament driven centrosome separation at G2-M transition by providing a bridge between the centrosome and actin-filaments. PMID- 18388322 TI - Adrenergic regulation of cardiac contractility does not involve phosphorylation of the cardiac ryanodine receptor at serine 2808. AB - The sympathetic nervous system is a critical regulator of cardiac function (heart rate and contractility) in health and disease. Sympathetic nervous system agonists bind to adrenergic receptors that are known to activate protein kinase A, which phosphorylates target proteins and enhances cardiac performance. Recently, it has been proposed that protein kinase A-mediated phosphorylation of the cardiac ryanodine receptor (the Ca(2+) release channel of the sarcoplasmic reticulum at a single residue, Ser2808) is a critical component of sympathetic nervous system regulation of cardiac function. This is a highly controversial hypothesis that has not been confirmed by several independent laboratories. The present study used a genetically modified mouse in which Ser2808 was replaced by alanine (S2808A) to prevent phosphorylation at this site. The effects of isoproterenol (a sympathetic agonist) on ventricular performance were compared in wild-type and S2808A hearts, both in vivo and in isolated hearts. Isoproterenol effects on L-type Ca(2+) current (I(CaL)), sarcoplasmic reticulum Ca(2+) release, and excitation-contraction coupling gain were also measured. Our results showed that isoproterenol caused significant increases in cardiac function, both in vivo and in isolated hearts, and there were no differences in these contractile effects in wild-type and S2808A hearts. Isoproterenol increased I(CaL), the amplitude of the Ca(2+) transient and excitation-contraction coupling gain, but, again, there were no significant differences between wild-type and S2808A myocytes. These results show that protein kinase A phosphorylation of ryanodine receptor Ser2808 does not have a major role in sympathetic nervous system regulation of normal cardiac function. PMID- 18388323 TI - Proteomic and metabolomic analysis of smooth muscle cells derived from the arterial media and adventitial progenitors of apolipoprotein E-deficient mice. AB - We have recently demonstrated that stem cell antigen 1-positive (Sca-1(+)) progenitors exist in the vascular adventitia of apolipoprotein E-deficient (apoE( /-)) mice and contribute to smooth muscle cell (SMC) accumulation in vein graft atherosclerosis. Using a combined proteomic and metabolomic approach, we now characterize these local progenitors, which participate in the formation of native atherosclerotic lesions in chow-fed apoE(-/-) mice. Unlike Sca-1(+) progenitors from embryonic stem cells, the resident Sca-1(+) stem cell population from the vasculature acquired a mature aortic SMC phenotype after platelet derived growth factor-BB stimulation. It shared proteomic and metabolomic characteristics of apoE(-/-) SMCs, which were clearly distinct from wild-type SMCs under normoxic and hypoxic conditions. Among the differentially expressed proteins were key enzymes in glucose metabolism, resulting in faster glucose consumption and a compensatory reduction in baseline interleukin-6 secretion. The latter was associated with a marked upregulation of insulin-like growth factor binding proteins (IGFBPs) 3 and 6. Notably, reconstitution of interleukin-6 to levels measured in the conditioned medium of wild-type SMCs attenuated the elevated IGFBP expression in apoE(-/-) SMCs and their vascular progenitors. This coregulation of apoE, interleukin-6, and IGFBPs was replicated in wild-type SMCs from hypercholesterolemic mice and confirmed by silencing apoE expression in SMCs from normocholesterolemic mice. In summary, we provide evidence that Sca-1(+) progenitors contribute to native atherosclerosis in apoE(-/-) mice, that apoE deficiency and hypercholesterolemia alter progenitor cell behavior, and that inflammatory cytokines such as interleukin-6 act as metabolic regulators in SMCs of hyperlipidemic mice. PMID- 18388324 TI - Long-term doxycycline is more effective than atenolol to prevent thoracic aortic aneurysm in marfan syndrome through the inhibition of matrix metalloproteinase-2 and -9. AB - Beta-blockers, eg, atenolol, are the cornerstone therapy for thoracic aortic aneurysm (TAA) in patients with Marfan syndrome; however, continued aortic dilatation has been reported. We have demonstrated that matrix metalloproteinase (MMP)-2 and -9 were upregulated during progression of TAA in Marfan syndrome, accompanied with degenerated elastic fibers and vasomotor dysfunction. We hypothesized that doxycycline, a nonspecific inhibitor of MMPs, would ameliorate TAA by attenuating elastic fiber degeneration and improving vasomotor function. A well-characterized mouse model of Marfan syndrome (Fbn1(C1039G/+)) was used. Mice were untreated (n=40), given doxycycline (0.24 g/L, n=30), or given atenolol (0.5 g/L, n=30) in drinking water at 6 weeks of age. The Fbn1(+/+) mice served as control (n=40). At 3, 6, and 9 months, aortic segments from the ascending, arch, and descending portions were used to obtain the "average" value of the whole thoracic aorta. TAA was prevented in the doxycycline group, whereas mild aneurysm was evident in the atenolol group. Doxycycline improved elastic fiber integrity, normalized aortic stiffness, and prevented vessel weakening. The impairment of vasocontraction and endothelium-dependent relaxation in the untreated and atenolol groups were improved by doxycycline. The upregulation of transforming growth factor-beta in the Marfan aorta was suppressed by doxycycline. Doxycycline augmented expression ratios of tissue inhibitors of MMP to MMPs. Intraperitoneally injected neutralizing antibodies against MMP-2 and -9 yielded similar effects to doxycycline. We concluded that long-term treatment with doxycycline, through the inhibition of MMP-2 and -9, is more effective than atenolol in preventing TAA in Marfan syndrome by preserving elastic fiber integrity, normalizing vasomotor function, and reducing transforming growth factor-beta activation. PMID- 18388325 TI - IP3 constricts cerebral arteries via IP3 receptor-mediated TRPC3 channel activation and independently of sarcoplasmic reticulum Ca2+ release. AB - Vasoconstrictors that bind to phospholipase C-coupled receptors elevate inositol 1,4,5-trisphosphate (IP(3)). IP(3) is generally considered to elevate intracellular Ca(2+) concentration ([Ca(2+)](i)) in arterial myocytes and induce vasoconstriction via a single mechanism: by activating sarcoplasmic reticulum (SR)-localized IP(3) receptors, leading to intracellular Ca(2+) release. We show that IP(3) also stimulates vasoconstriction via a SR Ca(2+) release-independent mechanism. In isolated cerebral artery myocytes and arteries in which SR Ca(2+) was depleted to abolish Ca(2+) release (measured using D1ER, a fluorescence resonance energy transfer-based SR Ca(2+) indicator), IP(3) activated 15 pS sarcolemmal cation channels, generated a whole-cell cation current (I(Cat)) caused by Na(+) influx, induced membrane depolarization, elevated [Ca(2+)](i), and stimulated vasoconstriction. The IP(3)-induced I(Cat) and [Ca(2+)](i) elevation were attenuated by cation channel (Gd(3+), 2-APB) and IP(3) receptor (xestospongin C, heparin, 2-APB) blockers. TRPC3 (canonical transient receptor potential 3) channel knockdown with short hairpin RNA and diltiazem and nimodipine, voltage-dependent Ca(2+) channel blockers, reduced the SR Ca(2+) release-independent, IP(3)-induced [Ca(2+)](i) elevation and vasoconstriction. In pressurized arteries, SR Ca(2+) depletion did not alter IP(3)-induced constriction at 20 mm Hg but reduced IP(3)-induced constriction by approximately 39% at 60 mm Hg. [Ca(2+)](i) elevations and constrictions induced by endothelin 1, a phospholipase C-coupled receptor agonist, were both attenuated by TRPC3 knockdown and xestospongin C in SR Ca(2+)-depleted arteries. In summary, we describe a novel mechanism of IP(3)-induced vasoconstriction that does not occur as a result of SR Ca(2+) release but because of IP(3) receptor-dependent I(Cat) activation that requires TRPC3 channels. The resulting membrane depolarization activates voltage-dependent Ca(2+) channels, leading to a myocyte [Ca(2+)](i) elevation, and vasoconstriction. PMID- 18388326 TI - An acyltransferase controls the generation of hematopoietic and endothelial lineages in zebrafish. AB - Hematopoietic and endothelial cells develop from a common progenitor, the hemangioblast, or directly from mesodermal cells. The molecular pathway that regulates the specification of both cell lineages remains elusive. Here, we show that a lysocardiolipin acyltransferase, lycat, is critical for the establishment of both hematopoietic and endothelial lineages. We isolated lycat from the deletion interval of cloche, a zebrafish mutant that has dramatically reduced hematopoietic and endothelial cell lineages. Reduction of lycat mRNA levels in wild-type zebrafish embryos decreases both endothelial and hematopoietic lineages. Lycat mRNA rescues blood lineages in zebrafish cloche mutant embryos. E165R and G166L mutations in the highly conserved catalytic domain in lycat abolish its function in zebrafish hematopoiesis. Epistasis analysis supports that lycat acts upstream of scl and etsrp in zebrafish hemangioblast development. These data indicate that lycat is the earliest known player in the generation of both endothelial and hematopoietic lineages. PMID- 18388327 TI - A novel RANTES antagonist prevents progression of established atherosclerotic lesions in mice. AB - BACKGROUND: Atherosclerosis is a chronic inflammatory disease that represents the primary cause of death through coronary disease and stroke. Chemokines are known to play a crucial role in this disease by recruiting inflammatory leukocytes to the endothelium. Recently, the chemokine variant [44AANA47]-RANTES was shown to impair inflammatory cell recruitment in vivo by interfering with heparin binding and oligomerization. METHODS AND RESULTS: In this study we report that curative treatment with [44AANA47]-RANTES limits atherosclerotic plaque formation in LDLr /- mice. This was associated with reduced infiltration of T cells and macrophages and reduced production of matrix metalloproteinase (MMP)-9. By contrast, the relative smooth muscle cell and collagen content was increased, indicating a more stable plaque phenotype. In addition, we provide evidence for direct inhibition of leukocyte recruitment into aortic root lesions, attenuated leukocyte rolling and arrest in mesenteric vessels, as well as a reduced proinflammatory response following Con A stimulation in vitro. CONCLUSIONS: Interference with chemokine oligomerization and chemokine/heparin interactions is a powerful novel approach that inhibits progression of established atherosclerosis in mice. By inhibiting leukocyte recruitment into plaques, [44AANA47]-RANTES mediates a less inflammatory plaque phenotype and thus reduced systemic inflammatory state. PMID- 18388328 TI - Regulation of endogenous apolipoprotein E secretion by macrophages. AB - Apolipoprotein E has critical roles in the protection against atherosclerosis and is understood to follow the classical constitutive secretion pathway. Recent studies have indicated that the secretion of apoE from macrophages is a regulated process of unexpected complexity. Cholesterol acceptors such as apolipoprotein A I, high density lipoprotein, and phospholipid vesicles can stimulate apoE secretion. The ATP binding cassette transporter ABCA1 is involved in basal apoE secretion and in lipidating apoE-containing particles secreted by macrophages. However, the stimulation of apoE secretion by apoA-I is ABCA1-independent, indicating the existence of both ABCA1-dependent and -independent pathways of apoE secretion. The release of apoE under basal conditions is also regulated, requiring intact protein kinase A activity, intracellular calcium, and an intact microtubular network. Mathematical modeling of apoE turnover indicates that whereas some pools of apoE are committed to either secretion or degradation, other pools can be diverted from degradation toward secretion. Targeted inhibition or stimulation of specific apoE trafficking pathways will provide unique opportunities to regulate the biology of this important molecule. PMID- 18388329 TI - Aliskiren-binding increases the half life of renin and prorenin in rat aortic vascular smooth muscle cells. AB - OBJECTIVE: Renin inhibition with aliskiren has been reported to cause a greater rise in renin than other types of renin-angiotensin system blockade, thereby potentially leading to angiotensin generation or stimulation of the human (pro)renin receptor (h(P)RR). Here we studied whether this rise in renin is attributable to an aliskiren-induced change in the prorenin conformation, allowing its detection in renin assays, or a change in renin/prorenin clearance. We also investigated whether aliskiren affects (pro)renin binding to its receptors, using rat aortic vascular smooth muscle cells (VSMCs) overexpressing the h(P)RR. METHODS AND RESULTS: A 48-hour incubation with aliskiren at 4 degrees C converted the prorenin conformation from "closed" to "open," thus allowing its recognition in active site-directed renin assays. VSMCs accumulated (pro)renin through binding to mannose 6-phosphate receptors (M6PRs) and h(P)RRs. Aliskiren did not affect binding at 4 degrees C. At 37 degrees C, aliskiren increased (pro)renin accumulation up to 40-fold, and M6PR blockade prevented this. Aliskiren increased the intracellular half life of prorenin 2 to 3 times. CONCLUSIONS: Aliskiren allows the detection of prorenin as renin, and decreases renin/prorenin clearance. Both phenomena may contribute to the "renin" surge during aliskiren treatment, but because they depend on aliskiren binding, they will not result in angiotensin generation. Aliskiren does not affect (pro)renin binding to its receptors. PMID- 18388330 TI - CCN1 knockdown suppresses neointimal hyperplasia in a rat artery balloon injury model. AB - OBJECTIVE: CCN1 (Cyr61) is an extracellular matrix-associated protein involved in cell proliferation and survival. CCN1 is bound to vascular smooth muscle cells (VSMCs) via integrins and is expressed in VSMCs in atherosclerotic lesions, suggesting involvement in the regulation of vascular smooth muscle cell (VSMC) proliferation and atherosclerosis. We hypothesized that knockdown of CCN1 may inhibit VSMC proliferation and suppress neointimal hyperplasia. METHODS AND RESULTS: We examined the effect of the knockdown of CCN1 using rat cultured VSMCs and a rat balloon injury model. CCN1 stimulated adhesion and migration of VSMCs in a dose-dependent manner, and this was blocked by an antibody for integrin alpha(6)beta(1). Moreover, knockdown of endogenous CCN1 by lentiviral delivery of siRNA significantly inhibited proliferation of VSMCs and the uptake of 5-bromo-2' deoxyuridine (BrdU). Replenishment with recombinant CCN1 reversed the effect of siRNA knockdown. Interestingly, knockdown of CCN1 significantly suppressed neointimal hyperplasia in a rat carotid artery balloon injury model at days 14 and 28 after injury. Gene transfer of CCN1 to smooth muscle reversed the effect of CCN1 knockdown on neointimal formation. These results suggest that endogenous CCN1 regulates proliferation of VSMCs and neointimal hyperplasia. CONCLUSIONS: Inhibition of CCN1 may provide a promising strategy for the prevention of restenosis after vascular interventions. PMID- 18388331 TI - Microvascular injury, thrombosis, inflammation, and apoptosis in the pathogenesis of heatstroke: a study in baboon model. AB - OBJECTIVE: Severe heatstroke is a leading cause of morbidity and mortality during heat waves. The pathogenesis of tissue injury, organ failure, and death in heatstroke is not well understood. METHODS AND RESULT: We investigated the pathways of heatstroke-induced tissue injury and cell death in anesthetized baboons (Papio hamadyras) subjected to environmental heat stress until core temperature attained 42.5 degrees C (moderate heatstroke; n = 3) or onset of severe heatstroke (n = 4) signaled by a fall in systolic blood pressure to < 90 mm Hg and rise in core temperature to 43.1+/-0.1 degrees C. Three sham-heated animals served as controls. Light and electron microscopy revealed widespread hemorrhage and thrombosis, transmural migration of leukocytes, and microvascular endothelium injury in severe heatstroke. Immunohistology and ultrastructural analysis demonstrated increased staining of endothelial von Willebrand factor (vWF), tissue factor (TF), and endothelial leukocyte-platelet interaction. Extensive apoptosis was noted in spleen, gut, and lung, and in hematopoeitic cells populating these organs. Double-labeling studies colocalized active caspase 3 and TF with apoptotic cells. Findings in sham-heated animals were unremarkable. CONCLUSIONS: These data suggested that microvascular injury, thrombosis, inflammation, and apoptosis may play an important role in the pathogenesis of heatstroke injury. PMID- 18388333 TI - Correlation between carotid intraplaque hemorrhage and clinical symptoms. PMID- 18388332 TI - Leukocyte telomere length and carotid artery intimal medial thickness: the Framingham Heart Study. AB - BACKGROUND AND PURPOSE: Leukocyte telomere length (LTL) is relatively short in individuals who have evidence of cardiovascular disease. The purpose of this study was to examine the link between LTL and the predisposition to atherosclerosis, as determined by carotid artery intimal medial thickness (IMT) in participants of the Framingham Offspring Study. METHODS AND RESULTS: LTL was assayed by the mean length of the terminal restriction fragments and carotid artery IMT by B-mode ultrasonography in 1062 individuals (496 men, 566 women) aged 33 to 86 years. In the whole sample, there was a significant association of age-and sex-adjusted LTL with internal carotid artery IMT (ICA-IMT) (r = -0.07, P = 0.02). In sex-stratified analysis, this association remained significant for men (r = -0.11, P = 0.02) but not for women (r = -0.04, P = 0.36). After further adjustment for cigarette smoking and BMI, a borderline significant association persisted in men (P = 0.06). In secondary analysis, the age-adjusted LTL was significantly (and negatively) associated with ICA-IMT (r = -0.28, p = 0.0006) in obese (BMI > 30 kg/m(2)) men but not in nonobese (BMI < or = 30 kg/m(2)) men. In addition, age-adjusted LTL was significantly shorter in men (6.89+/-0.02 kb) than women (7.01+/-0.02 kb; P < 0.0009) and in current cigarette smokers (6.87+/-0.05 kb) than never smokers (6.99+/-0.03 kb; P = 0.0006). Although there was no significant association of LTL with common carotid artery-IMT or with carotid artery stenosis, there was a significant inverse association of LTL with common carotid artery IMT in obese men. CONCLUSIONS: In obese men, shortened LTL is a powerful marker of increased carotid IMT. Given the public health impact of atherosclerosis and in particular the current epidemic of obesity, the associations noted in obese men warrant further confirmation. PMID- 18388334 TI - Detecting paroxysmal atrial fibrillation after ischemic stroke and transient ischemic attack: if you don't look, you won't find. PMID- 18388335 TI - Interventional management of stroke. PMID- 18388336 TI - Regional differences in diabetes as a possible contributor to the geographic disparity in stroke mortality: the REasons for Geographic And Racial Differences in Stroke Study. AB - BACKGROUND AND PURPOSE: Diabetes and hypertension impart approximately the same increased relative risk for stroke, although hypertension has a larger population attributable risk because of its higher population prevalence. With a growing epidemic of obesity and associated increasing prevalence of diabetes that disproportionately impacts the southeastern Stroke Belt states, any potential contribution of diabetes to the geographic disparity in stroke mortality will only increase. METHODS: Racial and geographic differences in diabetes prevalence and diabetes awareness, treatment, and control were assessed in the REasons for Geographic And Racial Differences in Stroke study, a national population-based cohort of black and white participants older than 45 years of age. At the time of this report, 21 959 had been enrolled. RESULTS: The odds of diabetes were significantly increased in both white and black residents of the stroke buckle (OR, 1.26; [1.10, 1.44]; OR, 1.45 [1.26, 1.66], respectively) and Stroke Belt (OR, 1.22; [1.09, 1.36]; OR, 1.13 [1.02, 1.26]) compared to the rest of the United States. In the buckle, regional differences were not fully mediated and remained significant when controlling for socioeconomic status and risk factors. Addition of hypertension to the models did not reduce the magnitude of the associations. There were no significant differences by region with regard to awareness, treatment, or control for either race. CONCLUSIONS: These analyses support a possible role of regional variation in the prevalence of diabetes as, in part, an explanation for the regional variation in stroke mortality but fail to support the potential for a contribution of regional differences in diabetes management. PMID- 18388337 TI - Natural history of dural arteriovenous shunts. AB - BACKGROUND AND PURPOSE: Dural arteriovenous shunts with cortical venous reflux or drainage may cause neurological symptoms and death with or without intracranial hemorrhage. Present knowledge about the natural history of these lesions is limited, however. We investigated the incidences of intracranial hemorrhage, progressive dementia syndrome, and death in patients diagnosed in our neurovascular center. METHODS: We evaluated the records of 85 patients with dural arteriovenous shunts with cortical venous drainage or reflux hospitalized in our institution from 1978 to 2007. The annual incidences of intracranial hemorrhage, progressive dementia syndrome, and death were calculated. RESULTS: Fifty-three patients did not have an intracranial hemorrhage as the presenting event. One of these patients bled after diagnosis. Thirty-two patients had an intracranial hemorrhage as the presenting event. Three patients bled after diagnosis. One of these patients died. Apart from deficits caused by hemorrhage, no patient reported adverse neurological symptoms. In patients presenting with an intracranial hemorrhage the annual risk for hemorrhage is approximately 7.4% and in those not presenting with a hemorrhage it is approximately 1.5%. CONCLUSIONS: The risk of intracranial hemorrhage from a dural arteriovenous shunt with cortical venous drainage is most likely smaller than previously proposed. Presentation with hemorrhage is a risk factor for hemorrhage. The risks of developing neurological symptoms not related to hemorrhage are also less than previously reported. PMID- 18388338 TI - Recanalization of an acute middle cerebral artery occlusion using a self expanding, reconstrainable, intracranial microstent as a temporary endovascular bypass. AB - BACKGROUND AND PURPOSE: Although self-expanding intracranial microstents have been used to treat acute middle cerebral artery (MCA) stroke, there are disadvantages associated with placing a permanent endovascular implant. We describe a technique in which a reconstrainable stent was used to provide a temporary endovascular bypass to achieve MCA recanalization without permanent stent implantation. METHODS: A 55-year-old male presented with acute onset left hemiplegia (National Institutes of Health Stroke Score (NIHSS) of 20. Angiography showed an occluded right cervical internal carotid artery (ICA), a patent anterior communicating artery (ACOMM), and embolic occlusion of the right middle cerebral artery (MCA), M1 segment. RESULTS: Working through a 6F guide-catheter positioned in the left cervical ICA, an SL-10 microcatheter, and 0.014-inch Synchro-2 microwire were manipulated across the anterior communicating artery and into the right M1 segment occlusion. 5 mg of abciximab and 3 mg tPA were infused directly into the thrombus through the microcatheter. Mechanical thrombolysis using the microwire and microcatheter was ineffective in achieving any recanalization. An Enterprise stent (4x22 mm) was delivered across the occlusion site and partially unconstrained. The unconstrained portion of the stent expanded and acted as a temporary bypass, to circumferentially displace and structurally disrupt the M1 thrombus, producing immediate revascularization of the right territory MCA. After approximately 20 minutes, the Enterprise stent was reconstrained and removed. Final angiography demonstrated excellent filling of the right M1 and distal MCA branches. The patient improved to an NIHSS of 7, regaining movement of his left upper and lower extremities. CONCLUSIONS: The temporary endovascular bypass technique yielded immediate and durable revascularization of an acutely occluded middle cerebral artery without the disadvantages associated with the placement of a permanent endovascular stent. PMID- 18388341 TI - Dual antithrombotic therapy increases severe bleeding events in patients with stroke and cardiovascular disease: a prospective, multicenter, observational study. AB - BACKGROUND AND PURPOSE: We sought to determine the incidence and severity of bleeding events in patients with stroke and cardiovascular diseases who were taking oral antithrombotic agents in Japan, where the incidence of hemorrhagic stroke is higher than in Western countries. METHODS: A prospective, multicenter, observational study was conducted; 4009 patients who were taking oral antithrombotic agents for stroke and cardiovascular diseases were enrolled. The patients were classified into 4 groups according to their antithrombotic treatment: the single antiplatelet agent group (47.2%); the dual antiplatelet agent group (8.7%); the warfarin group (32.4%); and the warfarin plus antiplatelet agent group (11.7%). The primary end point was life-threatening or major bleeding according to the MATCH trial definition. RESULTS: During a median follow-up of 19 months, there were 57 life-threatening and 51 major bleeding events, including 31 intracranial hemorrhages. The annual incidence of the primary end point was 1.21% in the single antiplatelet agent group, 2.00% in the dual antiplatelet agent group, 2.06% in the warfarin group, and 3.56% in the warfarin plus antiplatelet agent group (P<0.001). After adjustment for baseline characteristics, adding an antiplatelet agent to warfarin increased the risk of the primary end point (relative risk=1.76; 95% CI, 1.05 to 2.95), and adding another antiplatelet agent to single antiplatelet agent therapy increased the secondary end point of any bleeding, including minor events (relative risk=1.37; 95% CI, 1.07 to 1.76). CONCLUSIONS: The incidence of bleeding events during antithrombotic therapy in Japan was similar to that reported for Western countries, although the trials used different study designs. Dual antithrombotic therapy was independently related to an increased risk of bleeding events. PMID- 18388342 TI - Outpatient practice patterns after stroke hospitalization among neurologists. AB - BACKGROUND AND PURPOSE: Care after stroke hospitalization can provide several opportunities to optimize vascular risk reduction. However, not much is known about poststroke practice patterns among neurologists. Such knowledge may help direct specific efforts to improve the impact of practicing neurologists on clinical outcomes after stroke. METHODS: A survey soliciting information on processes of care in the outpatient setting after recent hospitalization for ischemic stroke or transient ischemic attack was mailed to a random sample of 833 US and Canadian neurologist-members of the American Academy of Neurology. RESULTS: A total of 475 (57%) responses were received. Practice demographics of survey responders and nonresponders were largely similar. Fourteen percent of respondents identified themselves as vascular neurologists. Overall, respondents reported frequently checking for medication adherence and counseling patients on lifestyle modification. However, neurologists reported screening more frequently for diabetes, hypertension, and dyslipidemia than actually treating these conditions (all P<0.0001) Vascular neurologists were more likely than general neurologists to screen for hypertension (97% versus 86%, P=0.016), dyslipidemia (94% versus 68%, P<0.001), diabetes (89% versus 62%, P<0.001), and sleep apnea (94% versus 79%, P=0.007) as well as to treat hypertension (71% versus 45%, P<0.001), dyslipidemia (82% versus 50%, P<0.001), diabetes (45% versus 21%, P<0.001), and current smoking (77% versus 59%, P=0.005). Neurologists with mostly government-insured and uninsured patients were significantly more likely to engage in vascular risk reduction treatment than neurologists with mostly commercially insured patients. CONCLUSIONS: Self-reported rates of screening and treatment of major vascular risk factors by most neurologists after stroke hospitalization are substantial but not universal. Bridging knowledge gaps or adopting a systematic management approach in coordination with primary care physicians could help optimize poststroke care. PMID- 18388343 TI - Granulocyte-macrophage colony-stimulating factor enhances leptomeningeal collateral growth induced by common carotid artery occlusion. AB - BACKGROUND AND PURPOSE: Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been reported to accelerate collateral growth (arteriogenesis) at the circle of Willis in rat brain. However, the effect of GM-CSF on leptomeningeal collateral growth has not been established. We examined the effect of unilateral common carotid artery (CCA) occlusion and GM-CSF treatment on leptomeningeal collateral growth in mice. METHODS: Adult mice were subjected to unilateral CCA occlusion or sham surgery followed by an alternate-day regimen of GM-CSF (20 microg/kg) or saline injection. On day 7, latex perfusion was performed in 1 set of mice to visualize the leptomeningeal vessels, and the number of Mac-2(+) monocytes/macrophages on the dorsal surface of the brain was counted. In another set of mice, on day 7, permanent ipsilateral middle cerebral artery (MCA) occlusion was performed, and infarct volume was measured. RESULTS: Leptomeningeal collateral growth was observed after CCA occlusion, and that was enhanced by GM CSF treatment. An increase in the number of Mac-2(+) cells on the surface of the brain occurred after CCA occlusion and was enhanced by GM-CSF treatment. Seven days after CCA occlusion, GM-CSF treatment decreased the infarct size attributable to subsequent MCA occlusion. CONCLUSIONS: After CCA occlusion, GM CSF treatment enhanced leptomeningeal collateral growth and decreased the infarct size after MCA occlusion in mice. PMID- 18388340 TI - Sarpogrelate-Aspirin Comparative Clinical Study for Efficacy and Safety in Secondary Prevention of Cerebral Infarction (S-ACCESS): A randomized, double blind, aspirin-controlled trial. AB - BACKGROUND AND PURPOSE: The antiplatelet agent sarpogrelate is a selective inhibitor of 5-hydroxytryptamine receptors. The purpose of this study was to compare the efficacy and safety of sarpogrelate with those of aspirin in Japanese ischemic stroke patients. METHODS: In total, 1510 patients with recent cerebral infarction (1 week to 6 months after onset) were randomly assigned to receive either sarpogrelate (100 mg TID) or aspirin (81 mg/d). Mean follow-up period was 1.59 years. The primary efficacy end point was recurrence of cerebral infarction. Clusters of serious vascular events (stroke, acute coronary syndrome, or vascular event-related death) were selected as secondary end points. The aim of the primary efficacy analysis was to demonstrate the noninferiority of sarpogrelate with respect to aspirin, with the criterion that the upper limit of the 95% CI of the hazard ratio (sarpogrelate vs aspirin) for recurrence of cerebral infarction should not exceed 1.33. RESULTS: Cerebral infarction recurred in 72 patients (6.09%/y) in the sarpogrelate group and in 58 (4.86%/y) in the aspirin group (hazard ratio=1.25; 95% CI, 0.89 to 1.77; P=0.19). A serious vascular event occurred in 90 (7.61%/y) and in 85 (7.12%/y) patients, respectively (hazard ratio=1.07; 95% CI, 0.80 to 1.44; P=0.65). The overall incidences of bleeding events were 89 (11.9%) and 131 (17.3%), respectively (P<0.01). CONCLUSIONS: Sarpogrelate was not noninferior to aspirin for prevention of recurrence of cerebral infarction. Bleeding events were significantly fewer with sarpogrelate than aspirin. The effect of aspirin in Japanese patients was similar to that in Western studies. PMID- 18388344 TI - Importance of in-hospital initiation of therapies and therapeutic inertia in secondary stroke prevention: IMplementation of Prevention After a Cerebrovascular evenT (IMPACT) Study. AB - BACKGROUND AND PURPOSE: Many patients do not receive prevention consistent with recommendations after stroke, but the relative importance of patient- and physician-related factors is uncertain. METHODS: We prospectively assessed factors associated with blood pressure (BP) <140/90 mm Hg and low-density lipoprotein (LDL) cholesterol <1 g/L in a collaborative cohort of 240 consecutive patients experiencing stroke/transient ischemic attack (Rankin <4; /=140/90 mm Hg, approximately 40% received either no treatment or one drug only, and treatment was reinforced in 20% of them only. Results were similar at 12 months with no improvement in the rate of control of risk factors. CONCLUSIONS: Therapeutic inertia is an important impediment to achieve BP and LDL control goals after stroke, even in fairly motivated/adherent patients. In hospital initiation of preventive therapies could improve quality of secondary stroke prevention in the long term. PMID- 18388346 TI - New brain lesions after carotid stenting versus carotid endarterectomy: a systematic review of the literature. AB - BACKGROUND AND PURPOSE: Against the background of a relatively low rate of clinical events during carotid angioplasty and stenting (CAS) or carotid endarterectomy (CEA), diffusion-weighted imaging (DWI) is increasingly being used to compare the incidence of new ischemic lesions after both procedures. In addition, DWI may also provide a means of defining the role of different CAS techniques on this adverse outcome. Therefore, we performed a PubMed search and systematically analyzed all peer-reviewed studies published between January 1990 and June 2007 reporting on the occurrence of new DWI lesions after CAS or CEA. Summary of Review- In 32 studies comprising 1363 CAS and 754 CEA procedures, the incidence of any new DWI lesion was significantly higher after CAS (37%) than after CEA (10%) (P<0.01). Similar results were obtained in a meta-analysis focusing on those studies directly comparing the incidence of new DWI lesions after either CEA or CAS (OR, 6.1; 95% CI, 4.19 to 8.87; P<0.01). The use of cerebral protection devices (33% vs 45% without; P<0.01) and closed-cell designed stents during CAS (31% vs 51% with open-cell stents; P<0.01), as well as selective versus routine shunt usage during CEA (6% vs 16%; P<0.01) significantly reduced the incidence of new ipsilateral DWI lesions. CONCLUSIONS: New DWI lesions occur more frequently after CAS than after CEA. However, technical advances mainly in the field of endovascular therapy potentially reduce the incidence of these adverse ischemic events. In this scenario, DWI appears to be an ideal tool to compare and further improve both techniques. PMID- 18388345 TI - MRI findings in the painful poststroke shoulder. AB - BACKGROUND AND PURPOSE: We describe the structural abnormalities in the painful shoulder of stroke survivors and their relationships to clinical characteristics. Method- Eighty-nine chronic stroke survivors with poststroke shoulder pain underwent T1- and T2-weighted multiplanar, multisequence MRI of the painful paretic shoulder. All scans were reviewed by one radiologist for the following abnormalities: rotator cuff, biceps and deltoid tears, tendinopathies and atrophy, subacromial bursa fluid, labral ligamentous complex abnormalities, and acromioclavicular capsular hypertrophy. Clinical variables included subject demographics, stroke characteristics, and the Brief Pain Inventory Questions 12. The relationship between MRI findings and clinical characteristics was assessed through logistic regression. RESULTS: Thirty-five percent of subjects exhibited a tear of at least one rotator cuff, biceps or deltoid muscle. Fifty-three percent of subjects exhibited tendinopathy of at least one rotator cuff, bicep or deltoid muscle. The prevalence of rotator cuff tears increased with age. However, rotator cuff tears and rotator cuff and deltoid tendinopathies were not related to severity of poststroke shoulder pain. In approximately 20% of cases, rotator cuff and deltoid muscles exhibited evidence of atrophy. Atrophy was associated with reduced motor strength and reduced severity of shoulder pain. CONCLUSIONS: Rotator cuff tears and rotator cuff and deltoid tendinopathies are highly prevalent in poststroke shoulder pain. However, their relationship to shoulder pain is uncertain. Atrophy is less common but is associated with less severe shoulder pain. PMID- 18388347 TI - Ubiquitin-mediated stress response in the spinal cord after transient ischemia. AB - BACKGROUND AND PURPOSE: Vulnerability of motor neurons in the spinal cord against ischemia is considered to play an important role in the development of delayed paraplegia after surgery of the thoracic aorta. However, the reasons for such vulnerability are not fully understood. Recently, the ubiquitin system has been reported to participate in neuronal cell death. In the present study, we investigated the expression of ubiquitin system molecules and discussed the relationship between the vulnerability and the ubiquitin system after transient ischemia in the spinal cord. METHODS: Fifteen minutes of spinal cord ischemia in rabbits was applied with the use of a balloon catheter. In this model, the spinal motor neuron shows selectively delayed neuronal death, whereas other spinal neurons such as interneurons survive. Immunohistochemical analysis and Western blotting for ubiquitin system molecules, ubiquitin, deubiquitylating enzyme (ubiquitin carboxy-terminal hydrolase 1), and ubiquitin-ligase parkin were examined. RESULTS: In cytoplasm, ubiquitin and ubiquitin carboxy-terminal hydrolase 1 were strongly induced both in interneuron and motor neuron at the early stage of reperfusion, but the sustained expression was observed only in motor neuron. Parkin was induced strongly at 3 hours after the reperfusion, but the immunoreactivity returned to the sham control level at 6 hours in both neurons. In the nuclei, ubiquitin, ubiquitin carboxy-terminal hydrolase 1, and parkin were strongly induced in interneuron, whereas no upregulation of these proteins was observed in motor neuron. CONCLUSIONS: These results indicate that the vulnerability of motor neuron of the spinal cord might be partially attributed to the different response in ubiquitin-mediated stress response after transient ischemia. PMID- 18388348 TI - Geographic prevalence and multilevel determination of community-level factors associated with herpes simplex virus type 2 infection in Chennai, India. AB - Herpes simplex virus type 2 (HSV-2) is one of the most prevalent sexually transmitted infections, and it increases the risk of transmission of human immunodeficiency virus type 1 at least twofold. Individual-level factors are insufficient to explain geographic and population variation in HSV-2, suggesting the need to identify ecologic factors. The authors sought to determine the geographic prevalence and community-level factors associated with HSV-2 after controlling for individual-level factors among slums in Chennai, India. From March to June 2001, participants aged 18-40 years voluntarily completed a survey and were tested for HSV-2. Community characteristics were assessed through interviews with key informants and other secondary data sources. Multilevel nonlinear analysis was conducted. Eighty-five percent of eligible persons completed the survey; of these, 98% underwent HSV-2 testing, producing a final sample of 1,275. Participants were of Tamil ethnicity, were predominantly female and married, and were on average 30 years old. Fifteen percent were infected with HSV-2, and there was significant variation in HSV-2 prevalence among communities. After controlling for individual-level factors, the authors identified community level factors, including socioeconomic status and the presence of injection drug users, that were independently associated with HSV-2 and explained 11% of the variance in prevalence. Future studies are needed to test mechanisms through which these community-level factors may be operating. PMID- 18388349 TI - Src as a potential therapeutic target in non-small-cell lung cancer. AB - Lung cancer is the most common cause of cancer-related death, with non-small-cell lung cancer (NSCLC) accounting for 80%-85% of all cases. Although survival rates are reasonably good for patients diagnosed with very early disease, the majority of patients present with advanced disease. For these patients, palliation and improvements in quality of life are the primary goals of therapy. Although chemotherapeutic agents remain the cornerstone of first-line therapy, these agents have limited use in patients who have relapsed and have metastatic disease. Therefore, new strategies are required to improve survival and quality of life in this setting. With the substantial advances in our understanding of tumour biology, it has been possible to identify signalling pathways involved in mediating tumour growth and progression. These pathways offer targets for new biological agents such as small molecule inhibitors and monoclonal antibodies. One such target is Src, a tyrosine kinase that is involved in multiple aspects of tumorigenesis including proliferation, migration and angiogenesis. Increased levels of Src expression have been found in a range of cancers, especially breast, colorectal, prostate and lung. Preliminary preclinical data and pharmacodynamic data suggest that Src inhibition is a viable therapeutic option in the treatment of advanced NSCLC. PMID- 18388350 TI - Listening to musical rhythms recruits motor regions of the brain. AB - Perception and actions can be tightly coupled; but does a perceptual event dissociated from action processes still engage the motor system? We conducted 2 functional magnetic resonance imaging studies involving rhythm perception and production to address this question. In experiment 1, on each trial subjects 1st listened in anticipation of tapping, and then tapped along with musical rhythms. Recruitment of the supplementary motor area, mid-premotor cortex (PMC), and cerebellum was observed during listen with anticipation. To test whether this activation was related to motor planning or rehearsal, in experiment 2 subjects naively listened to rhythms without foreknowledge that they would later tap along with them. Yet, the same motor regions were engaged despite no action-perception connection. In contrast, the ventral PMC was only recruited during action and action-coupled perceptual processes, whereas the dorsal part was only sensitive to the selection of actions based on higher-order rules of temporal organization. These functional dissociations shed light on the nature of action-perception processes and suggest an inherent link between auditory and motor systems in the context of rhythm. PMID- 18388351 TI - Zoledronic acid is effective against experimental malignant pleural effusion. AB - RATIONALE: Aminobiphosphonates, such as zoledronic acid (ZA), exert potent indirect antitumor effects and are currently being tested against human solid tumors. The antitumor actions of aminobiphosphonates, including angiostasis, are relevant to the pathogenesis of malignant pleural effusion (MPE), but no study has addressed the efficacy of these compounds against malignant pleural disease. OBJECTIVES: Here we hypothesized that treatment of immunocompetent mice with ZA would halt tumor progression in a mouse model of adenocarcinoma-induced MPE. METHODS: To induce MPE in mice, Lewis lung carcinoma cells were delivered directly into the pleural space. Subsequently, animals were treated with ZA in both a prevention and a regression protocol. MEASUREMENTS AND MAIN RESULTS: ZA treatment resulted in significant reductions in pleural fluid accumulation and tumor dissemination, while it significantly prolonged survival. These effects of ZA were linked to enhanced apoptosis of pleural tumor cells, decreased formation of new vessels in pleural tumors, and reduced pleural vascular permeability. In addition, ZA was able to inhibit the recruitment of mononuclear cells to pleural tumors, with concomitant reductions in matrix metalloproteinase-9 release into the pleural space. Finally, ZA limited the expression of proinflammatory and angiogenic mediators, as well as the activity of small GTP proteins Ras and RhoA, in tumor cells in vivo and in vitro. CONCLUSIONS: ZA is effective against experimental MPE, suggesting that this intervention should be considered for testing in clinical trials. PMID- 18388352 TI - Tuberculosis outbreaks predicted by characteristics of first patients in a DNA fingerprint cluster. AB - RATIONALE: Some clusters of patients who have Mycobacterium tuberculosis isolates with identical DNA fingerprint patterns grow faster than others. It is unclear what predictors determine cluster growth. OBJECTIVES: To assess whether the development of a tuberculosis (TB) outbreak can be predicted by the characteristics of its first two patients. METHODS: Demographic and clinical data of all culture-confirmed patients with TB in the Netherlands from 1993 through 2004 were combined with DNA fingerprint data. Clusters were restricted to cluster episodes of 2 years to only detect newly arising clusters. Characteristics of the first two patients were compared between small (2-4 cases) and large (5 or more cases) cluster episodes. MEASUREMENTS AND MAIN RESULTS: Of 5,454 clustered cases, 1,756 (32%) were part of a cluster episode of 2 years. Of 622 cluster episodes, 54 (9%) were large and 568 (91%) were small episodes. Independent predictors for large cluster episodes were as follows: less than 3 months' time between the diagnosis of the first two patients, one or both patients were young (<35 yr), both patients lived in an urban area, and both patients came from sub-Saharan Africa. CONCLUSIONS: In the Netherlands, patients in new cluster episodes should be screened for these risk factors. When the risk pattern applies, targeted interventions (e.g., intensified contact investigation) should be considered to prevent further cluster expansion. PMID- 18388353 TI - Idiopathic nonspecific interstitial pneumonia: report of an American Thoracic Society project. AB - RATIONALE: The 2002 American Thoracic Society/European Respiratory Society classification of idiopathic interstitial pneumonias identified nonspecific interstitial pneumonia (NSIP) as a provisional diagnosis. Concern was expressed that NSIP was a "wastebasket" category, difficult to distinguish from other idiopathic interstitial pneumonias. OBJECTIVES: The following questions were addressed: (1) Is idiopathic NSIP a distinct entity? 2) If so, what are its clinical, radiologic and pathologic characteristics? (3) What is the role of radiology and pathology in establishing the diagnosis? (4) To make a diagnosis of idiopathic NSIP, what other disorders need to be excluded and how should this be done? METHODS: Investigators who had previously reported cases of idiopathic NSIP were invited to submit cases for review (n = 305). After initial review, cases with complete clinical, radiologic, and pathologic information (n = 193) were reviewed in a series of workshops. MEASUREMENTS AND MAIN RESULTS: Sixty-seven cases were identified as NSIP. Mean age was 52 years, 67% were women, 69% were never-smokers, and 46% were from Asian countries. The most common symptoms were dyspnea (96%) and cough (87%); 69% had restriction. By high-resolution computed tomography, the lower lung zones were predominantly involved in 92% of cases; 46% had a peripheral distribution; 47% were diffuse. Most showed a reticular pattern (87%) with traction bronchiectasis (82%) and volume loss (77%). Lung biopsies showed uniform thickening of alveolar walls with a spectrum of cellular to fibrosing patterns. Five-year survival was 82.3%. CONCLUSIONS: Idiopathic NSIP is a distinct clinical entity that occurs mostly in middle-aged women who are never smokers. The prognosis of NSIP is very good. PMID- 18388354 TI - Stimulation of lung innate immunity protects against lethal pneumococcal pneumonia in mice. AB - RATIONALE: The lungs are a common site of serious infection in both healthy and immunocompromised subjects, and the most likely route of delivery of a bioterror agent. Since the airway epithelium shows great structural plasticity in response to inflammatory stimuli, we hypothesized it might also show functional plasticity. OBJECTIVES: To test the inducibility of lung defenses against bacterial challenge. METHODS: Mice were treated with an aerosolized lysate of ultraviolet-killed nontypeable (unencapsulated) Haemophilus influenzae (NTHi), then challenged with a lethal dose of live Streptococcus pneumoniae (Spn) delivered by aerosol. MEASUREMENTS AND MAIN RESULTS: Treatment with the NTHi lysate induced complete protection against challenge with a lethal dose of Spn if treatment preceded challenge by 4 to 24 hours. Lesser levels of protection occurred at shorter (83% at 2 h) and longer (83% at 48-72 h) intervals between treatment and challenge. There was also some protection when treatment was given 2 hours after challenge (survival increased from 14 to 57%), but not 24 hours after challenge. Protection did not depend on recruited neutrophils or resident mast cells and alveolar macrophages. Protection was specific to the airway route of infection, correlated in magnitude and time with rapid bacterial killing within the lungs, and was associated with increases of multiple antimicrobial polypeptides in lung lining fluid. CONCLUSIONS: We infer that protection derives from stimulation of local innate immune mechanisms, and that activated lung epithelium is the most likely cellular effector of this response. Augmentation of innate antimicrobial defenses of the lungs might have therapeutic value. PMID- 18388355 TI - Hepatotoxicity of pyrazinamide: cohort and case-control analyses. AB - RATIONALE: Relatively little is known about the hepatotoxicity of pyrazinamide. OBJECTIVES: We compared continuation-phase regimens incorporating pyrazinamide, isoniazid, and/or rifampin with those containing isoniazid and rifampin to evaluate the hepatotoxicity of pyrazinamide. METHODS: Cohort and nested case control analyses were conducted on a cohort of 3,007 patients with active tuberculosis (TB) managed at government chest clinics under a TB control program with treatment started from January 1 through June 30, 2001. Cases included all patients with probable hepatotoxicity from 12 or more weeks after starting treatment. Hepatotoxicity was considered probable when serum alanine transaminase exceeded three times the upper limit of normal. Each case was matched by sex and age with three control subjects selected randomly from the rest of the cohort. Treatment regimens of cases within 4 weeks preceding hepatotoxicity were compared with those of matched control subjects in comparable periods relative to the date of commencing treatment. MEASUREMENTS AND MAIN RESULTS: Hepatotoxicity occurred in 150 (5.0%) patients at any time including 48 (1.6%) cases. From 12 or more weeks after starting treatment, the estimated risk of hepatotoxicity was 2.6% for regimens incorporating pyrazinamide, isoniazid, and/or rifampin, and 0.8% for standard regimens containing isoniazid and rifampin. Multivariable conditional logistic analysis showed a significant association between hepatotoxicity and, respectively, hepatitis B, previous hepatotoxicity, and treatment regimens. The adjusted odds ratio (95% confidence interval) of hepatotoxicity for regimens incorporating pyrazinamide, isoniazid, and/or rifampin relative to standard regimens was 2.8 (1.4-5.9). CONCLUSIONS: Adding pyrazinamide to isoniazid and rifampin increases the risk of hepatotoxicity appreciably. PMID- 18388356 TI - Acetazolamide for Monge's disease: efficiency and tolerance of 6-month treatment. AB - RATIONALE: Monge's disease is characterized by an excessive erythrocytosis, frequently associated with pulmonary hypertension, in high-altitude dwellers. It has a considerable impact on public health in high-altitude regions. A preliminary study demonstrated the efficiency of acetazolamide (Acz) (250 mg/d for 3 wk) in reducing serum erythropoietin and hematocrit. OBJECTIVES: Evaluate the efficacy and tolerance of a 6-month treatment with 250 mg Acz that could be chronically implemented and its effects on pulmonary artery pressure and cardiac function. METHODS: A two-phase study was performed in patients (hematocrit > or = 63%) from Cerro de Pasco, Peru (4,300 m). First phase: a double-blind, placebo controlled study in 55 patients who received a single dose of either 250 mg Acz (n = 40) or placebo (n = 15) by daily oral administration for 12 weeks. Second phase (open label): after a 4-week washout period, all patients received 250 mg Acz for 12 weeks. Hematocrit, blood gases, clinical outcome, and pulmonary artery circulation were evaluated. MEASUREMENTS AND MAIN RESULTS: First phase: Acz decreased by 44% the number of polycythemic subjects (P = 0.02), decreased hematocrit from 69 to 64% (P < 0.001), and increased arterial O(2) pressure from 42 to 45 mm Hg (P < 0.001). No severe adverse effect or hypokalemia was recorded. The second phase reproduced the effects observed during the first phase, without cumulative effects on hematocrit. A 4-week washout restored basal hematocrit. Only patients who received Acz for 6 months showed a clear reduction in pulmonary vascular resistance. CONCLUSIONS: Acz reduces erythrocytosis and improves pulmonary circulation in Monge's disease without adverse effects. Its implementation as a chronic treatment for this disease appears efficient and safe. PMID- 18388358 TI - Incidence and prognosis of sustained arrhythmias in critically ill patients. AB - RATIONALE: Sustained arrhythmias are common in postoperative and cardiac intensive care units (ICUs), but their incidence and prognosis in general ICUs have never been reported. OBJECTIVES: To estimate the incidence and prognosis of sustained arrhythmias in a general ICU population. METHODS: Prospective, multicenter, 1-month inception cohort study. MEASUREMENTS AND MAIN RESULTS: A total of 1,341 patients were included: 12% (163/1,341) had sustained arrhythmias, including 8% (113/1,341) and 2% (30/1,341) with supraventricular and ventricular arrhythmias, respectively, and 2% (30/1,341) with conduction abnormalities. In hospital death rates were 17% (205/1,178) in patients without arrhythmia and 29% (33/113) in patients with supraventricular arrhythmias (odds ratio [OR], 1.95; 95% confidence interval [CI], 1.27-3.01), 73% (22/30) in patients with ventricular arrhythmias (OR, 13.20; 95% CI, 5.79-30.10), and 60% (18/30) in patients with conduction abnormalities (OR, 7.46; 95% CI, 3.52-15.82). Neurological sequel rates were 6% (55/973) in arrhythmia-free survivors and 15% (12/80) in survivors with supraventricular arrhythmias (OR, 2.92; 95% CI, 1.45 5.89), 38% (3/8) in survivors with ventricular arrhythmias (OR, 7.53; 95% CI, 1.60-35.50), and 17% (2/12) in survivors with conduction abnormalities (OR, 8.77; 95% CI, 1.65-46.57). After adjusting for prognosis factors and propensity scores, ventricular arrhythmias still increased mortality (OR, 3.53; 95% CI, 1.19-10.42) but supraventricular arrhythmias and conduction abnormalities did not. CONCLUSIONS: Sustained arrhythmias are observed in 12% of patients admitted to general ICUs. Ventricular arrhythmias increase the risk of death. PMID- 18388357 TI - Endotoxin augments myeloid dendritic cell influx into the airways in patients with allergic asthma. AB - RATIONALE: Epidemiologic studies have shown that exacerbation of asthma is modulated by environmental endotoxin. High levels of endotoxin are associated with asthma symptoms and the current use of asthma medication. However, the underlying mechanisms by which endotoxin modulates asthma are not completely understood. OBJECTIVES: The aim of the study was to test whether endotoxin enhances the response of individuals with allergic asthma to allergen, and to determine if this interaction is associated with increased numbers of antigen presenting cells in the airways. METHODS: Seventeen subjects with mild allergic asthma underwent segmental challenge with allergen, endotoxin, and the combination of both in three different lung segments via bronchoscopy. The cellular influx including monocytes, myeloid dendritic cells (mDCs), and plasmacytoid dendritic cells (pDCs), as well as the level of cytokines, were assessed in bronchoalveolar lavage fluid obtained 24 hours after segmental challenge. Monocytes, mDCs, and pDCs were isolated and their capacity to induce T cell proliferation was determined. MEASUREMENTS AND MAIN RESULTS: Endotoxin enhanced the cellular response to allergen. The combination of allergen and endotoxin resulted in increased numbers of total cells, lymphocytes, neutrophils, eosinophils, monocytes, and mDCs, as well as increased levels of lipopolysaccharide-binding protein, IL-1alpha, IL-6, and tumor necrosis factor alpha in the bronchoalveolar lavage fluid compared with allergen alone. Isolated mDCs but not pDCs induced a strong T cell proliferation in vitro. CONCLUSIONS: Endotoxin augments the allergic inflammation in the lungs of individuals with asthma, and induces an enhanced influx of monocytes and functionally active antigen-presenting mDCs into the respiratory tract. PMID- 18388359 TI - Up-regulation of the glutathione S-transferase system in human liver by polycyclic aromatic hydrocarbons; comparison with rat liver and lung. AB - The cytosolic glutathione S-transferases (GSTs) comprise a pivotal enzyme system protecting the cell from electrophilic compounds. It plays a major role in the detoxication of the primary and dihydrodiol epoxides of polycyclic aromatic hydrocarbons (PAHs), so that modulation of this enzyme system by PAHs will impact on their carcinogenic activity. The potential of six structurally diverse PAHs, namely benzo[a]pyrene (B[a]P), fluoranthene, benzo[b]fluoranthene (B[b]F), dibenzo[a,l]pyrene, dibenzo[a,h]anthracene (D[a,h]A) and 1-methhylphenanthrene, to modulate hepatic GST activity was investigated in human precision-cut slices and compared to rat slices, a species frequently used in long-term carcinogenicity studies; changes were monitored at the activity, using three different substrates, protein and mRNA levels. When activity was monitored using the alpha-class selective 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, B[b]F was the only PAH that caused an increase in activity, which was accompanied by a rise in the Ya immunoreacting band. In rat slices, in addition to B[b]F, B[a]P and D[a,h]A also enhanced activity, being paralleled with increased levels of the Ya immunoreacting band. In the rat, all PAHs elevated mRNA levels. In both human and rat liver slices, only B[b]F enhanced activity when 1-chloro-2,4-dinitrobenzene (CDNB) served as substrate. To investigate tissue differences, similar studies were undertaken in precision-cut rat lung slices, incubated with PAHs under identical conditions, using CDNB, as this was the only substrate for which activity could be detected; none of the PAHs studied stimulated activity. It is concluded that some PAHs have the potential to induce GST activity in human liver tissue and that species and tissue differences exist in the induction of this enzyme system in the rat. However, the extent of induction of GST activity is very modest compared with the effect these compounds have on CYP1 expression, the family responsible for their bioactivation, and it is unlikely to compensate for the enhanced production of reactive intermediates. PMID- 18388360 TI - The sensitivity and specificity of thermometry and plethysmography in the assessment of hand-arm vibration syndrome. AB - BACKGROUND: Finger plethysmography and thermometry are objective measures used to assess the vascular aspect of hand-arm vibration syndrome (HAVS). Research to date shows poor correlation between these tests and Stockholm Workshop Scale (SWS) vascular stage. Clinicians, researchers and compensation boards require objective means to diagnose and quantify HAVS. AIMS: To define the specificity and sensitivity of thermometry and plethysmography using the SWS as the reference criterion. A secondary goal was to consider cut points for the tests optimizing sensitivity and specificity. METHODS: A cross-sectional analysis was conducted on HAVS patients seen at an occupational medicine specialty clinic. Plethysmography and thermometry were analyzed using SWS vascular stage as the outcome variable. Logistic regression controlled for age, smoking and time since last vibration exposure and use of vasoactive medications. The sensitivity and specificity of the combined tests were calculated using varying cut points. RESULTS: A total of 139 patients consented to participate in the study. Plethysmography stage 1 or greater showed the highest sensitivity (sensitivity 94% and specificity 15%). Specificity was optimized combining plethysmography stage 3 and thermometry stage 3 (specificity 98% and sensitivity 23%). Maximal diagnostic accuracy was achieved by plethysmography alone setting the criteria for a positive test as being stage 1 or greater (70%). CONCLUSIONS: Neither plethysmography nor thermometry either alone or in combination demonstrated sufficient sensitivity and specificity to serve as an objective correlate for SWS vascular stage. All combinations of plethysmography and thermometry showed a lower specificity than sensitivity indicating that the SWS may be less sensitive in detecting vascular pathology than the objective tests. PMID- 18388361 TI - Management of autoimmune urticaria. PMID- 18388362 TI - Cosmetic dermatology versus cosmetology: a misnomer in need of urgent correction. PMID- 18388363 TI - Psoriasiform dermatoses. AB - Psoriasiform reaction pattern is a commonly encountered denominator in a wide variety of unrelated disorders. It may be a reaction to either the internal or the external environmental, allergic, infective, parasitic, bacterial, fungal, viral and/or malignant stimuli. The degree of evolution of such a pattern and its significance vary according to the dermatosis. The age of the skin lesions may also influence the histopathological presentation and its clinico histopathological disparity can often bewilder an expert. However, such a situation warrants more astute and sustained observations to unveil the exact underlying condition(s). Thus, psoriasiform dermatoses should only be an initial caption until an exact dermatological disorder is defined. There has been greater number of instances of psoriasiform drug eruptions where a confirmation of the diagnosis can be achieved after their remission by doing a provocation test. Similarly, such instances have also been on the rise in HIV/AIDS-affected individuals all over the world. Besides mycosis fungoides and Hodgkin's disease, several unrelated malignancies have been preceded or accompanied by psoriasiform skin eruptions. PMID- 18388364 TI - A study of allergen-specific IgE antibodies in Indian patients of atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic, relapsing, pruritic dermatitis frequently associated with the hyperproduction of IgE to various allergens. Identification of these allergens is possible by various laboratory investigations. AIM: The present study was designed to assess these allergen specific antibodies in the diagnosis of AD in the Indian context. METHODS: This prospective study comprised 50 patients of AD. The diagnosis was made clinically after satisfying Hanifin and Rajka's criteria. Serum IgE levels were estimated and specific IgE antibodies were measured for 20 food allergens and aeroallergens. RESULTS: Serum IgE was elevated in 88% of the patients. The highest elevation of mean IgE levels was seen in the 10-20 years age group. Sixty five percent of the children under the age of ten years were positive to one or more food allergens. Food allergens were more often positive in the < or = 10 years age group and specific antibodies to inhalants were seen more frequently in the older age groups. Specific antibodies to apples were found in all age groups. CONCLUSION: Antibodies against apples and hazelnuts were the more commonly seen specific antibodies in children. Incidence of positivity was much higher in children when compared to earlier studies. Identification of food allergens can be an important factor in the diagnosis of AD in children in India. Positivity to inhalant allergens in the older age groups was lower in this study. The allergen profile with regard to inhalants in Indian patients was similar to that of earlier studies. PMID- 18388365 TI - Chronic idiopathic urticaria: comparison of clinical features with positive autologous serum skin test. AB - BACKGROUND: Chronic idiopathic urticaria (CIU), in its extremely severe form, can pose a therapeutic challenge to the treating physician. It has been noted that in one third of such patients, autoantibodies against the IgE receptor are seen and such patients have more severe and unremitting urticaria. AIM: To compare clinical features of autoimmune urticaria with those of other CIU patients. METHODS: We conducted a prospective study in an attempt to correlate the clinical features with autoantibodies, indirectly detected via the autologous serum skin test (ASST), which is the simplest and the best in vivo clinical test for detection of basophil histamine-releasing activity. DISCUSSION: Out of 100 patients with chronic idiopathic urticaria, 34 showed a positive reaction to the autologous serum skin test and it was found that the frequency and severity of attacks was higher in these patients. CONCLUSION: ASST may be used as a simple and cost-effective test for the classification of chronic urticaria, which has proven to be a therapeutic challenge to the treating physician. PMID- 18388366 TI - Autologous serum therapy in chronic urticaria: old wine in a new bottle. AB - BACKGROUND: Chronic urticaria (CU) is one of the most challenging and frustrating therapeutic problems faced by a dermatologist. A recent demonstration of abnormal type 1 reactions to intradermal autologous serum injections in some CU patients has led to the characterization of a new subgroup of "autoimmune chronic urticaria". This has rekindled interest in the age-old practice of autologous blood injections as a theoretically sound treatment option in these patients. AIMS: To evaluate the efficacy of repeated autologous serum injections (ASIs) in patients with recalcitrant chronic urticaria. METHODS: A cohort of 62 (32 females) CU patients with a positive autologous serum skin test (ASST) (group 1) was prospectively analyzed for the efficacy of nine consecutive weekly autologous serum injections with a postintervention follow-up of 12 weeks. Another group of 13 (seven females) CU patients with negative ASST (group 2) was also treated similarly. In both groups, six separate parameters of disease severity and activity were recorded. RESULTS: Demographic and disease variables were comparable in both groups. The mean duration of disease was 1.9 +/- 0.3 years (range = 3 months to 32 years) in group 1 and 1.5 +/- 0.2 years (range = 3 months to 10 years) in group 2. In the ASST (+) group, 35.5% patients were completely asymptomatic at the end of the follow-up while an additional 24.2% were markedly improved. In the ASST (-) group, these figures were 23 and 23% respectively. The intergroup difference for complete subsidence was statistically significant (P < 0.05). In both groups, the most marked reduction was seen in pruritus and antihistamine use scores followed by the size and frequency of the wheals. CONCLUSION: Autologous serum therapy is effective in a significant proportion of ASST (+) patients with CU. A smaller but still substantial number of ASST (-) patients also benefited from this treatment. PMID- 18388367 TI - Use of patch testing for identifying allergen causing chronic urticaria. AB - BACKGROUND AND AIMS: The aim of this present study was to evaluate the role of patch testing for the etiological diagnosis of chronic urticaria (CU) by using the Indian standard battery of patch test allergens approved by Contact and Occupational Dermatitis Forum of India (CODFI). METHODS: A total of 57 cases with chronic urticaria were tested with the Indian standard battery of allergens. All those cases that showed allergy to patch test allergens were advised to avoid contact with the allergen(s) to whom they had allergy; they were also advised to avoid/restrict allergens in the diet. This avoidance/restriction was advised for a period of six weeks. During this period, clinical improvement of each patient was evaluated and recorded at weekly intervals. RESULTS: Out of the 57 cases of CU, 11 patients showed positive reactions to one or more patch test allergens. Nine out of eleven showed complete disappearance of CU by 2-3 weeks on avoidance of the allergen and this improvement continued till the end of six weeks. The remaining two cases showed partial recovery from CU during the same period. CONCLUSION: Patch testing is a safe, simple and inexpensive alternative that can be used for the etiological diagnosis of chronic urticaria before undertaking expensive investigations. PMID- 18388368 TI - Vitiligoid lichen sclerosus: a reappraisal. AB - BACKGROUND: Many case studies of lichen sclerosus (LS) have reported an association of vitiligo. However, such an association is not reported from larger case studies of vitiligo, which happens to be a common disease. Autoimmune etiology suspected in both LS and vitiligo has been considered as the reason for their association in some patients. It has also been suggested that lichenoid inflammation in LS may trigger an autoimmune reaction against melanocytes. AIMS: To test this association, we reviewed clinical and histological features of 266 cases of vitiligo and 74 cases of LS in a concurrent study of both diseases. METHODS: All outpatients seen in our department between 2003 and 2006 and who were diagnosed as having LS or vitiligo on the basis of clinical and pathologic features were included in the study. RESULTS: Vitiligoid lesions were seen along with stereotypical LS lesions in three patients but all the three lesions had histological features of LS. Oral/genital areas were affected in 57 out of the 74 LS cases and of those, 15 were initially suspected to have vitiligo. These cases with a clinical appearance of vitiligo and histological features of LS were considered as 'vitiligoid LS', a superficial variant proposed by J. M. Borda in 1968. Association of LS was not observed in the 266 cases of vitiligo. CONCLUSION: Exclusive oral/genital depigmentation is a common problem and histological evaluation is essential to differentiate vitiligoid LS from true vitiligo. The association of vitiligo with LS may have been documented due to the clinical misdiagnosis of vitiligoid LS lesions as vitiligo as histological investigations were not undertaken in any of the reported cases. PMID- 18388369 TI - Activated charcoal and baking soda to reduce odor associated with extensive blistering disorders. AB - BACKGROUND: Skin disease leading to extensive blistering and loss of skin is associated with a characteristic smell. Odor can cause physiologic disturbances such as increase in heart rate and respiratory rate. It can also cause nausea and vomiting and is disturbing to bystanders. AIMS: To test odor reducing capability of activated charcoal. METHODS: In this blinded experimental study we used putrefied amniotic membrane to produce odor and studied the effectiveness of activated charcoal and soda-bi-carbonate to reduce odor. RESULTS: Statistical analysis with Kruskal Wall's Chi Square Test and Man Whitney U test showed significant reduction of odor using activated charcoal by itself or along with soda-bi-carbonate. CONCLUSION: We recommend the usage of activated charcoal with/without soda bicarbonate as an inexpensive practical measure to reduce foul odor associated with extensive skin loss. PMID- 18388370 TI - Nevus of Ota: a series of 15 cases. AB - BACKGROUND: The nevus of Ota is a dermal nevus characterized by bluish pigmentation in the distribution of the first and the second division of the trigeminal nerve. AIM: Our aim was to study the cutaneous and extracutaneous manifestations of the nevus of Ota. METHODS: A total of 15 cases were included in our study. A detailed history, clinical examination along with direct ophthalmoscopy and otoscopy were done for all the cases. RESULTS: Most of the patients (60%) had lesions at birth and the majority (86.7 %) were females. Five (33.3%) patients belonged to Tanino class II. Combined dermal and ocular involvement was observed in 60% of the cases. CONCLUSION: Tanino class II was the most common type observed in our studies. A few rarer associations such as nevus of Ito and hemangioma were also noted in our patients. PMID- 18388371 TI - Premature ovarian failure due to cyclophosphamide: a report of four cases in dermatology practice. AB - Immunosuppressant drugs like cyclophosphamide are used in the treatment of a variety of skin disorders. Though it is a very useful drug, it has some serious side-effects. Prolonged amenorrhea due to premature ovarian failure leading to infertility is one of the serious side-effects of cyclophosphamide. Four cases of cyclophosphamide-induced premature ovarian failure are presented. Two patients of scleroderma, one patient of pemphigus and one patient of hypersensitivity vasculitis developed amenorrhea due to premature ovarian failure leading to infertility after receiving cyclophosphamide 50 mg o.d. for eight months to one year. The ages of these patients ranged from 28-38 years. All these patients had good improvement of their disease with cyclophosphamide. These patients did not experience any other side-effects and their routine blood and urine tests were normal. There were no spontaneous menses during the follow-up period of one to two years. Because of the serious risk of developing premature ovarian failure, cyclophosphamide should be avoided in those patients where the family is not complete. PMID- 18388372 TI - Hand, foot and mouth disease in Nagpur. AB - Hand, foot and mouth disease (HFMD) is a viral infection of children caused by Coxsackie virus-A16, a type of enterovirus closely related with the virus that causes herpangina. Although seen worldwide, it is not common in India. Hand, foot and mouth disease is sporadically reported from India as a mild illness. This report describes four cases of HFMD from Nagpur, Central India, seen between September 2005 and April 2006. All patients presented with a mild febrile prodrome followed by the appearance of aphthous-like oral ulcers and vesicular lesions on the hands and feet. All cases were clinically diagnosed. Coxsackie virus A16 was isolated from the serum of one of the patients. All the patients were in the age group of 3-5 years from different schools. It was a mild illness and all the four patients recovered without any complication. There were no secondary cases in the families. PMID- 18388373 TI - Non-familial multiple keratoacanthomas in a 70 year-old long-term non-progressor HIV-seropositive man. AB - We describe here multiple keratoacanthomas in an Human Immunodeficiency Virus (HIV)-seropositive 70 year-old man. The patient had multiple epithelial tumors of the skin showing rapid growth, histopathological features of a keratoacanthoma and a conspicuous tendency toward spontaneous remission. A diagnosis of nonfamilial multiple keratoacanthoma was established. The patient had a CD4 count of 633 cells/microL. The HIV disease in our patient was of a nonprogressive nature with CCR5-positive T cells. PMID- 18388374 TI - Late onset isotretinoin resistant acne conglobata in a patient with acromegaly. AB - A 55 year-old male presented with multiple pus-discharging abscesses and sinuses and mutilating scarring on the gluteal region and back prevalent for the last ten years with exacerbations and remissions. Physical examination revealed acromegaly with frontal bossing, prognathism, a barrel chest and acral hypertrophy. Dermatological examination revealed cutis verticis gyrata, thick eyelids, a large triangular nose, a thickened lower lip, macroglossia, widely spaced teeth and widened skin pores with wet and oily skin. Hair was fine and nails were flat and wide. There were multiple inflammatory papules, tender nodules, draining sinuses, and grouped, polyporous comedones as well as multiple and extensive depressed and keloidal scars localized predominantly over the gluteal region with a few scattered lesions over the back. A computed tomography (CT) scan showed widened sella turcica. His basal fasting growth hormone (GH) levels were markedly raised (230 ng/mL; normal 1-5 ng/mL) while the prolactin levels were moderately raised (87 ng/mL; normal 2-5 ng/mL). These findings were consistent with a diagnosis of acromegaly. The patient was put on antibiotics, nonsteroidal antiinflammatory drugs and isotretinoin at a dose of 1 mg/kg/day, which was increased to 1.5 mg/kg/day. Except for an initial mildly beneficial response, the skin lesions were largely resistant to high doses of isotretinoin at the end of four months. PMID- 18388375 TI - Familial dyskeratotic comedones. AB - Familial dyskeratotic comedones (FDC) is a rare autosomal dominant inherited condition, characterized by widespread, symmetrically scattered, comedone-like, hyperkeratotic papules, which are cosmetically unappealing. These lesions appear around puberty and show a predilection to involve the trunk, arms and face. The lesions are asymptomatic and gradually worsen with time. Histology shows invagination of the epidermis with a lamellar keratinous plug and focal evidence of dyskeratosis. This condition is generally refractory to therapy. We report here two cases with this rare disorder who had a strong familial history of the same disorder. PMID- 18388376 TI - Nasal NK/T cell lymphoma presenting as a lethal midline granuloma. AB - Nasal NK/T cell lymphomas are aggressive, locally destructive, midfacial, necrotizing lesions. The nonspecific clinical symptoms constitute a major stumbling block in the early diagnosis and management of these lymphomas. We report here a case of probable nasal NK/T cell lymphoma in an apparently healthy male that progressed rapidly in a short span of time and was managed subsequently with chemotherapy and external beam irradiation with which the lesion regressed. PMID- 18388377 TI - Childhood sclerodermatomyositis with generalized morphea. AB - Systemic sclerosis (SS) and dermatomyositis (DM) are both multisystem disorders and share some common clinical features. We report here an 11 year-old girl whose disease showed a changing clinical pattern from juvenile systemic sclerosis (JSS) to slowly progressing juvenile dermatomyositis (JDM) and had associated generalized morphea. Serological studies revealed antinuclear antibodies (ANA) with a speckled pattern. Topoisomerase-I (Scl-70), U1 RNP (ribonucleoprotein), anti-Ro, anti-La and anti Jo-1 antibody tests were negative. Electromyography (EMG) was suggestive of primary muscle disease and histopathological findings indicated scleroderma. The patient fulfilled the American College Rheumatology (ACR) diagnostic criteria for JSS as well as Bohan and Peter criteria for JDM separately and hence, was diagnosed to have sclerodermatomyositis (SDM). Mixed connective tissue disease (MCTD) and antisynthetase antibody syndrome (ASS) which share same clinical features with SS and DM were excluded by immunological studies. PMID- 18388378 TI - Subcutaneous panniculitis-like T-cell cutaneous lymphoma. AB - Subcutaneous panniculitis-like T cell lymphoma (SPTCL) is a rare cytotoxic T-cell lymphoma classified in the World Health Organization-European Organization for Research and Treatment of Cancer (WHO-EORTC) classification as a unique extranodal lymphoma with characteristic by T cell receptor (TCR) gene rearrangement. We report here a case of SPTCL in a 22 year-old woman who had presented with variably sized multiple nodules on both her legs. Initial differential diagnoses considered were panniculitis and lupus panniculitis. The histopathology showed a predominantly subcutaneous lobular infiltrate with atypical lymphocytes, karyorrhexis and rimming of adipocytes by lymphoid cells. Immunohistochemistry showed CD4-, CD8+, CD56- T-cell phenotype. Although TCR rearrangement studies were not done, the above T-cell phenotype and sparing of epidermis and dermis suggested the possibility of an SPTCL alpha/beta type. The patient received five cycles of a cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) regimen which resulted in the regression in her skin lesions and constitutional symptoms. PMID- 18388379 TI - Peutz-Jeghers syndrome with prominent palmoplantar pigmentation. PMID- 18388380 TI - Using a submersible pump to clean large areas of the body with antiseptics. PMID- 18388381 TI - Stratum corneum findings as clues to histological diagnosis of pityriasis lichenoides chronica. PMID- 18388382 TI - Omalizumab in severe chronic urticaria. PMID- 18388383 TI - Hypothesis: the potential utility of topical eflornithine against cutaneous leishmaniasis. PMID- 18388384 TI - Nodular melanoma in a skin graft site scar. PMID- 18388385 TI - Palatal involvement in lepromatous leprosy. PMID- 18388386 TI - Unilateral nevoid telangiectasia with no estrogen and progesterone receptors in a pediatric patient. PMID- 18388387 TI - Eruptive lichen planus in a child with celiac disease. PMID- 18388388 TI - Xerosis and pityriasis alba-like changes associated with zonisamide. PMID- 18388390 TI - Vitiligo, psoriasis and imiquimod: fitting all into the same pathway. PMID- 18388389 TI - Treatment of actinomycetoma with combination of rifampicin and co-trimoxazole. PMID- 18388391 TI - Multiple dermatofibromas on face treated with carbon dioxide laser: the importance of laser parameters. PMID- 18388392 TI - Alopecia areata progressing to totalis/universalis in non-insulin dependent diabetes mellitus (type II): failure of dexamethasone-cyclophosphamide pulse therapy. PMID- 18388393 TI - Subungual exostosis. PMID- 18388394 TI - Clinicohistopathological correlation of leprosy. PMID- 18388395 TI - Dermatographism. PMID- 18388396 TI - Mycophenolate mofetil. PMID- 18388397 TI - Multiple papules on the vulva. PMID- 18388398 TI - Functional analysis of alpha5beta1 integrin and lipid rafts in invasion of epithelial cells by Porphyromonas gingivalis using fluorescent beads coated with bacterial membrane vesicles. AB - Porphyromonas gingivalis, a periodontal pathogen, was previously suggested to exploit alpha5beta1 integrin and lipid rafts to invade host cells. However, it is unknown if the functional roles of these host components are distinct from one another during bacterial invasion. In the present study, we analyzed the mechanisms underlying P. gingivalis invasion, using fluorescent beads coated with bacterial membrane vesicles (MV beads). Cholesterol depletion reagents including methyl-beta-cyclodextrin (MbetaCD) drastically inhibited the entry of MV beads into epithelial cells, while they were less effective on bead adhesion to the cells. Bead entry was also abolished in CHO cells deficient in sphingolipids, components of lipid rafts, whereas adhesion was negligibly influenced. Following MbetaCD treatment, downstream events leading to actin polymerization were abolished; however, alpha5beta1 integrin was recruited to beads attached to the cell surface. Dominant-negative Rho GTPase Rac1 abolished cellular engulfment of the beads, whereas dominant-negative Cdc42 did not. Following cellular interaction with the beads, Rac1 was found to be translocated to the lipid rafts fraction, which was inhibited by MbetaCD. These results suggest that alpha5beta1 integrin, independent of lipid rafts, promotes P. gingivalis adhesion to epithelial cells, while the subsequent uptake process requires lipid raft components for actin organization, with Rho GTPase Rac1. PMID- 18388399 TI - Dynein-dependent movement of autophagosomes mediates efficient encounters with lysosomes. AB - Autophagy is a membrane trafficking pathway that carries cytosolic components to the lysosome for degradation. During this process, the autophagosome, a double membraned organelle, is generated de novo, sequesters cytoplasmic proteins and organelles, and delivers them to lysosomes. However, the mechanism by which autophagosomes are targeted to lysosomes has not been determined. Here, we observed the real-time behavior of microtubule-associated protein light chain 3 (LC3), which localizes to autophagosomes, and showed that autophagosomes move in a microtubule- and dynein-dynactin motor complex-dependent manner. After formation, autophagosomes show a rapid vectorial movement in the direction of the centrosome, where lysosomes are usually concentrated. Microinjection of antibodies against LC3 inhibited this movement; furthermore, using FRAP, we showed that anti-LC3 antibody injection caused a defect in targeting of autophagosomes to lysosomes. Collectively, our data demonstrate the functional significance of autophagosome movement that enables effective delivery from the cytosol to lysosomes. PMID- 18388400 TI - Comparative study on the vitamin C contents of the food legume seeds. AB - We found that dehydrated legume seeds (6 genera, 19 species and cultivated varieties) contained considerable amounts of vitamin C (VC). The average value of total VC content per 100 g of dry weight in dehydrated seeds varied from 0.24 mg (kidney beans) to 4.14 mg (green peas). Yard beans showed highest values among all legumes examined here in the both dehydrated and rehydrated forms (3.19 and 10.8 mg, respectively). By soaking for 16 h in the dark at 20(o)C, total VC contents of black grams and mung beans increased to 3.1- and 4.5-fold, respectively. However, three varieties of green peas (Hakuryu, Kurumeyutaka, and Nankaimidori) significantly lost their VC during the same soaking treatment. Total VC content of a rehydrated and cooked mung beans was higher than that of a dehydrated form. Appreciable amounts of total VC were detected in the immature seeds of six different genera such as yard beans, kidney beans, broad beans, green peas, soybeans and peanuts. Except for mung beans, 70-100% of VC in dehydrated seeds of adzuki beans, broad beans, green peas, black soybeans, and soybeans was lost by boiling. Total VC and L-ascorbic acid in mung beans, green peas, broad beans, black soybeans, and adzuki beans remained even after boiling, suggesting that it is possible to obtain VC from the cooked forms of these legume seeds. PMID- 18388401 TI - The direct precursor of the pyrimidine moiety of thiamin is not urocanic acid but histidine in Saccharomyces cerevisiae. AB - The biosynthetic route of the pyrimidine moiety of thiamin is different in prokaryotes and eukaryotes. In prokaryotes, the pyrimidine moiety is synthesized from aminoimidazole ribonucleotide, an intermediate of purine biosynthesis, while in eukaryotes, we have reported that the N-1, C-2, and N-3 atoms of the imidazole ring of histidine are incorporated into N-3, C-4, and the amino group attached to the C-4 atoms of the pyrimidine moiety, respectively, as a unit; the rest of the atoms of the pyrimidine moiety originate from pyridoxine as a unit. It has been reported that urocanic acid, the deaminated compound of histidine, is the direct precursor of the pyrimidine moiety. In the present report, we have investigated whether histidine or urocanic acid is the direct precursor of the pyrimidine moiety in Saccharomyces cerevisiae, using tracer experiments with 1) (13)C formate and urocanic acid, 2) (15)N-NH(4)Cl and urocanic acid, 3) (15)N-NH(4)Cl and histidine, and 4) (13)C-histidine and urocanic acid. The GC-MS analysis revealed that the incorporation of the (15)N atom of (15)NH(4)Cl was not affected by the presence of urocanic acid, although it was affected by histidine, and the incorporation of (13)C-histidine was not affected by the presence of urocanic acid. These results confirm that histidine is the direct precursor of the pyrimidine moiety of thiamin in S. cerevisiae. PMID- 18388402 TI - Beneficial effects of combined administration of alendronate and alfacalcidol on cancellous bone mass of the tibia in orchidectomized rats: a bone histomorphometry study. AB - The purpose of the present study was to examine the effects of combined administration of alendronate (ALN) and alfacalcidol (ALF) on the cancellous and cortical bone mass of the tibia in orchidectomized rats. Fifty male Sprague Dawley rats, 3 mo of age, were randomized by the stratified weight method into five groups: age-matched control, orchidectomy, and orchidectomy with administration of ALN (2.5 mug/kg, s.c., 5 times a week), ALF (0.05 microg/kg, p.o., 5 times a week), or ALN+ALF. The total experimental period was 12 wk. Orchidectomy reduced the cancellous bone mass of the proximal tibial metaphysis and maturation-related cortical bone gain of the tibial diaphysis as a result of increased trabecular bone resorption and decreased periosteal bone formation and also increased endocortical bone erosion and formation. ALN suppressed trabecular bone resorption and endocortical bone erosion and formation and increased periosteal bone formation, while ALF increased the number of osteoblasts and suppressed trabecular bone resorption and markedly increased periosteal and endocortical bone formation. Thus, both ALN and ALF prevented the orchidectomy induced reduction in the cancellous bone mass and maturation-related cortical bone gain. Combined administration of ALN and ALF increased the cancellous bone mass as compared with the values observed in age-matched controls by causing more marked suppression of trabecular bone resorption. The present study showed the beneficial effects of combined administration of ALN and ALF on the cancellous bone mass of the tibia in orchidectomized rats. PMID- 18388403 TI - Determination of individual vitamin B(6) compounds based on enzymatic conversion to 4-pyridoxolactone. AB - A determination method for individual natural vitamin B(6) compounds was developed. The vitamin B(6) compounds were specifically converted into 4 pyridoxolactone (PAL), a highly fluorescent compound, through a combination of enzymatic reactions and HCl-hydrolysis. PAL was then determined by HPLC. Pyridoxal was completely oxidized to PAL with pyridoxal 4-dehydrogenase (PLDH). Pyridoxine and pyridoxamine were totally converted into PAL through a coupling reaction involving pyridoxine 4-oxidase and PLDH, and one involving pyridoxamine pyruvate aminotransferase and PLDH, respectively. The 5'-phosphate forms and pyridoxine-beta-glucoside were hydrolyzed with HCl, and then determined as their free forms. Pyridoxine 5'-phosphate and pyridoxine-beta-glucoside were not separately determined here. Three food samples were analyzed by this method. PMID- 18388404 TI - Development of a simple food frequency questionnaire to estimate intakes of calcium and other nutrients for the prevention and management of osteoporosis. AB - There have been no simple methods to estimate dietary nutrient intakes for the prevention and management of osteoporosis. The aim of this study was to develop and validate a new, simple food frequency questionnaire (FFQ) for dietary intake of calcium and other nutrients relevant to the bone health of adult Japanese women. We developed a 28-item FFQ. To validate this, 208 and 72 adult women aged between 18 and 69 y were recruited for testing reliability and reproducibility, respectively. In the 208 women, moderate-to-high Spearman's correlation coefficients between our FFQ and the conventional diet record method were found in intakes of calcium (r=0.668), sodium chloride (NaCl) (r=0.475), vitamin A (r=0.501), vitamin D (r=0.413), vitamin K (r=0.649), and energy (r=0.471). In the 72 women, coefficients of variance of the four repeated measurements of intakes throughout a year were 14.1% for calcium, 7.3% for NaCl, 21.2% for vitamin A, 13.6% for vitamin D, 36.8% for vitamin K, and 9.6% for energy. In conclusion, the FFQ we developed is a useful tool to evaluate the intake of dietary calcium of adult Japanese women. Although it can also measure intakes of dietary vitamin A, vitamin D, vitamin K, NaCl, and energy, further improvement is needed to measure intakes of these nutrients and energy. PMID- 18388405 TI - In vitro diffusion barriers of the mouse jejunum in Ussing chambers. AB - Epithelial cells and their intercellular junctions play a primary role in the intestinal mucosal diffusion barrier to the invasion of noxious agents. Our previous study has shown that the mouse jejunum, when incubated in Ussing chambers for 4 h, exhibited morphological deterioration of the villi with denudation of the epithelia while it retained cAMP-induced potential difference (PD), suggesting that some subepithelial tissues had taken part in the barrier functions when the primary epithelial barrier on the villous surface was broken. To further characterize the barrier function of the jejunum in Ussing chambers, we measured the unidirectional lucifer yellow flux (J(LY)), which represents the permeation of a medium-sized anion, the transmural electrical conductance (G(t)), which reflects the permeation of electrolytes, mainly NaCl, and forskolin-induced PD. The values of J(LY), G(t) and forskolin-induced PD were not affected by removing the muscularis propria, suggesting that this tissue did not substantially contribute to the barrier. In addition, the values of J(LY), G(t) and forskolin-induced PD were not correlated with the degree of denudation of epithelial cells on the mucosal surface, supporting the notion that the main barrier function is constituted by the subepithelial tissues rather than the epithelium. Loosening the tissue (thereby approximating the free diffusion condition) by removing Ca(2+) from the bathing solution increased both J(LY) and G(t), with the former being larger than the latter. In conclusion, the mucosa propria and/or the submucosa constitute a diffusion barrier that restricted the permeation of lucifer yellow more tightly than NaCl in the injured jejunum incubated in Ussing chambers. PMID- 18388406 TI - Inter-modal comparisons of acute energy expenditure during perceptually based exercise in obese adults. AB - Previous studies have suggested that if exercise intensity is established by perceived effort, the metabolic demand varies among exercise machines and the treadmill optimizes energy expenditure (EE). However, these studies have been completed utilizing young people with normal body fat percentages. Therefore, the purpose of this study was to assess whether there was a difference in acute EE when obese people used different exercise modes at a self-selected intensity (ratings of perceived exertion 11-12) commonly recommended for overweight individuals. Twelve obese subjects (7 male; 5 female; BMI>29 kg/m(2)), aged 37-71 y completed two familiarization trials on four machines: treadmill (TM), stationary cycle (C), body trec elliptical arm/leg (BT), and airdyne (AD). On separate days, subjects then completed a 15 min trial on each machine at a self selected intensity corresponding to a target RPE of 11-12 on the Borg 15-point scale. Machine order was randomly assigned, and subjects were blinded to the workload throughout each trial. Workload was self-adjusted during the first 5 min and then remained stable for the rest of the trial. Physiological data were obtained during the last 5 min of each trial. The BT produced the highest rate of EE among exercise machines and C the lowest. These results suggest that perceptually-based exercise prescriptions are not reliable across modes typically found in a fitness center environment, and that weight-bearing arm/leg exercise optimizes EE during self-selected exercise of moderate intensity in obese subjects. PMID- 18388407 TI - Fish-oil high-fat diet intake of dams after day 5 of pregnancy and during lactation guards against excessive fat consumption of their weaning pups. AB - To investigate the influence of parental fat intake on preferential fat intake by pups after weaning, two groups of dams in study 1 were fed either a low-fat diet (LFD) or a lard high-fat diet (HFD) and those in study 2 were fed either a LFD or a fish-oil HFD after day 5 of pregnancy and during lactation. In study 1, when pups were placed on a self-selection regimen of the LFD and the lard HFD within the first week after weaning, the ratio of the lard HFD intake [lard HFD intake (g)/total intake (g)] by pups of both groups was about 70%. Although pups nursed by dams fed the lard HFD continued to eat the same ratio of the lard HFD, the ratio for pups nursed by dams fed the LFD gradually decreased to 20% in week 3 after weaning. In study 2, when pups were placed on a self-selection regimen of the LFD and the fish-oil HFD after weaning, the ratio of the fish-oil HFD intake in both groups of pups nursed by dams fed the LFD and the fish-oil HFD was about 20% for 3 wk after weaning. In studies 1 and 2, although no significant difference in dietary intake or body weight of dams and pups was observed among all groups through the experimental period, perirenal fat tissue weight of dams fed the lard HFD was higher than that of dams fed the LFD. These findings indicate that (1) fat preference of weaning pups nursed by dams fed the lard HFD is higher than that of weaning pups nursed by dams fed the LFD, and (2) intake of dam's fish-oil HFD diet guards against pups' intake of excessive fat. PMID- 18388408 TI - Zinc bioavailability is improved by the micronised dispersion of zinc oxide with the addition of L-histidine in zinc-deficient rats. AB - Zinc fortification of milk or soft drinks is usually used to combat zinc deficiencies in developing countries. Water-soluble zinc compounds, such as zinc sulfate or zinc citrate, are better absorbed but have an unacceptable taste. A micronised, dispersible zinc oxide (MDZnO), which does not have such a problem concerning taste, had higher solubility compared to ZnO (zinc oxide) in an artificial gastric solution. MDZnO was tested for its bioavailability using zinc deficient Wistar rats. Prior to the experiment, rats were fed zinc-deficient diet for 3 wk and were orally administered control (distilled water) or zinc solutions (ZnO, ZnO+L-histidine (His), MDZnO, MDZnO+His, 1 mg zinc/kg or 3.2 mg His/kg body weight). Compared to ZnO, MDZnO showed a lag in peak time and a lengthy period of continued high plasma zinc concentration after the single oral administration of zinc compounds. Addition of His to MDZnO elevated serum zinc concentration. Serum zinc concentration (area under the curve) in rats administered MDZnO with His was significantly higher than in rats administered distilled water (p<0.05). Liver zinc level was significantly higher in rats administered MDZnO with His compared with control rats (p<0.05), although the level was not affected by the administration with ZnO alone, ZnO+His, or MDZnO alone. In conclusion, the solubility of ZnO was elevated by the micronised dispersion tecnique and an in vivo study using zinc-deficient rats confirmed that its bioavailability was significantly improved compared to ZnO and the coadministration of His additively enhanced the bioavailability of MDZnO. PMID- 18388409 TI - Lipid peroxidation potential and antioxidants in the heart tissue of beta-alanine or taurine-treated old rats. AB - The aim of this study was to investigate the effect of the changes of taurine levels in the hearts of old rats on endogenous malondialdehyde (MDA) and diene conjugate (DC) levels and ascorbic acid (AA)- and NADPH-induced lipid peroxidation as well as non-enzymatic (glutathione, vitamin E and vitamin C) and enzymatic antioxidants (superoxide dismutase, glutathione peroxidase and glutathione transferase). Two groups of old (22 mo) rats were treated with beta alanine (3%, w/v; in drinking water), a taurine depleting agent, or taurine (2% w/v; in drinking water) for 6 wk. Significant decreases were observed in taurine contents of hearts in old rats as compared to young (5 mo) rats. We found that MDA and DC levels and AA- and NADPH-induced lipid peroxidation increased, but non enzymatic and enzymatic antioxidants did not alter in heart homogenates of aged rats. beta-Alanine administration resulted in significant decreases in heart taurine levels of old rats. This treatment did not cause further increases in MDA or DC levels or changes in antioxidants. However, AA- and NADPH-induced lipid peroxidation was higher than that of old rats. Taurine treatment caused significant increases in heart taurine levels of old rats. This treatment was found to decrease endogenous MDA and DC levels without affecting the antioxidant system in the heart homogenates of aged rats. AA- and NADPH-induced lipid peroxidation was also reduced in old rats when given taurine, although not statistically significantly. Our results indicate that the changes in heart taurine levels may influence the susceptibility of heart tissue to lipid peroxidation in aged rats and that taurine supplementation has protective effects on age-dependent oxidative stress in heart tissue. PMID- 18388410 TI - Effects of high magnesium intake on bone mineral status and lipid metabolism in rats. AB - Effects of magnesium (Mg)-intake on mineral levels in bone, blood and various tissues, bone density and plasma lipids were examined in 4-wk-old, female Wistar rats fed diets of five different Mg-contents of 1/10, 1 (standard diet (Mg 47 mg/100 g diet): control group), 2, 5 and 10-fold of the standard content for 4 wk. In rats fed the 1/10-fold Mg diet, the body weight gain declined with a statistical significance compared to control rats, while no significant differences were observed in rats fed on the 2-, 5-, and 10-fold Mg diets. The imbalance of mineral concentrations in the rats fed 1/10 Mg diet was induced, but in the rats fed high Mg diets were not induced except in tibia. Mg concentration in tibia of rats fed the 10-fold Mg diet significantly increased compared to the control group. All groups (1/10-fold, 2-, 5-, and 10-fold Mg groups) showed no significant changes in tibial mineral levels, except Mg in the 10-fold group, compared to the control group. With increase in Mg-intake, decline of plasma lipid parameters such as cholesterol, triglycerides and phospholipids are induced. Based on these results, it was suggested that a higher Mg-intake (around 5-fold of the standard Mg-content) might have a prophylactic potential against the onset of hyperlipidemia and cardiovascular disease by reducing lipid indices, and Mg decline in bone. PMID- 18388411 TI - The growth hormone affects the brain protein synthesis rate in hypophysectomized aged rats. AB - The purpose of this study was to determine whether the growth hormone (GH) affects the rate of brain protein synthesis in hypophysectomized aged rats. Experiments were conducted on three groups of 24-wk-old male rats: group 1 were hypophysectomized to reduce the level of plasma GH, group 2 were hypophysectomized and treated with GH and group 3 were sham-operated controls. The fractional rates of protein synthesis in the brains of hypophysectomized rats with GH were significantly greater than those in hypophysectomized rats without GH. In the cerebral cortex and cerebellum, the RNA activity [g protein synthesized/(g RNA.d)] significantly correlated with the fractional rate of protein synthesis (r>0.88, p<0.001). The RNA concentration (mg RNA/ g protein) was also related to the fractional rate of protein synthesis in these organs (r>0.56, p<0.05). The results suggest that the treatment of GH to hypophysectomized aged rats is likely to increase the rate of protein synthesis in the brain, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis. PMID- 18388412 TI - Chestnut astringent skin extract, an alpha-amylase inhibitor, retards carbohydrate absorption in rats and humans. AB - Inhibitors of carbohydrate-hydrolyzing enzyme play an important role to control postprandial blood glucose levels. In this paper, we investigated the effect of an ethanol extract from chestnut astringent skin (CAS) on alpha-amylase. Chestnut astringent skin extract strongly inhibited human and porcine pancreatic alpha amylase. We also investigated the effect of CAS extract on carbohydrate absorption in rats and humans. Oral administration of CAS extract to normal rats fed corn starch (2 g/kg body weight), significantly suppressed the increase of blood glucose levels after starch loading in a dose-dependent manner. The effective dose of CAS extract required to achieve 20 and 40% suppression of the rise in blood glucose level was estimated to be 40 and 155 mg/kg body weight, respectively. Chestnut astringent skin extract also suppressed the rise in plasma insulin level and the fall in plasma non-esterified fatty acid level. In the type 2 diabetic rat model, CAS extract significantly suppressed the rise in blood glucose level after starch loading in a dose-dependent manner. Chestnut astringent skin extract also suppressed the rise in plasma glucose level after boiled rice loading in a dose-dependent manner in humans. The amount of CAS extract required to achieve 11 and 23% suppression in the rise in plasma glucose level was 300 and 600 mg/person, respectively. These results suggest that CAS extract retards absorption of carbohydrate and reduces post-prandial hyperglycemia. PMID- 18388413 TI - Effect of green tea on volatile sulfur compounds in mouth air. AB - Many food products are claimed to be effective in controlling halitosis. Halitosis is caused mainly by volatile sulfur compounds (VSCs) such as H(2)S and CH(3)SH produced in the oral cavity. Oral microorganisms degrade proteinaceous substrates to cysteine and methionine, which are then converted to VSCs. Most treatments for halitosis focus on controlling the number of microorganisms in the oral cavity. Since tea polyphenols have been shown to have antimicrobial and deodorant effects, we have investigated whether green tea powder reduces VSCs in mouth air, and compared its effectiveness with that of other foods which are claimed to control halitosis. Immediately after administering the products, green tea showed the largest reduction in concentration of both H(2)S and CH(3)SH gases, especially CH(3)SH which also demonstrated a better correlation with odor strength than H(2)S; however, no reduction was observed at 1, 2 and 3 h after administration. Chewing gum, mints and parsley-seed oil product did not reduce the concentration of VSCs in mouth air at any time. Toothpaste, mints and green tea strongly inhibited VSCs production in a saliva-putrefaction system, but chewing gum and parsley-seed oil product could not inhibit saliva putrefaction. Toothpaste and green tea also demonstrated strong deodorant activities in vitro, but no significant deodorant activity of mints, chewing gum or parsley-seed oil product were observed. We concluded that green tea was very effective in reducing oral malodor temporarily because of its disinfectant and deodorant activities, whereas other foods were not effective. PMID- 18388414 TI - Short-term effects of glucosyl hesperidin and hesperetin on blood pressure and vascular endothelial function in spontaneously hypertensive rats. AB - Glucosyl hesperidin (G-hesperidin) is a water-soluble derivative of hesperidin. In the present study, the short-term effects of G-hesperidin and hesperetin, a putative metabolite of G-hesperidin, in spontaneously hypertensive rats (SHR) and in normotensive Wistar-Kyoto rats (WKY) were investigated. Single oral administration of G-hesperidin (10 to 50 mg/kg) induced a dose-dependent reduction in systolic blood pressure (SBP) in SHR, but had no effects in WKY. Intraperitoneal injection of hesperetin (50 mg/kg) into SHR also caused a significant reduction in SBP. The depressor effect was significantly inhibited by a nitric oxide synthase inhibitor, N(G)-nitro-L-arginine methyl ester. Moreover, hesperetin (10(-5) M) enhanced endothelium-dependent relaxation induced by acetylcholine, but had no effect on endothelium-independent relaxation induced by sodium nitroprusside in isolated aortas from SHR. These data suggest that the hypotensive effect of hesperetin in SHR is associated with nitric oxide-mediated vasodilation. Therefore, this effect may be involved in the mechanisms by which G hesperidin lowers blood pressure in hypertension. PMID- 18388415 TI - Protective effect of lipoic acid against methylglyoxal-induced oxidative stress in LLC-PK(1) cells. AB - Methylglyoxal (MG), a reactive dicarbonyl compound, is a metabolic byproduct of glycolysis often found at high levels in blood from diabetic patients. The effect of lipoic acid on MG-induced oxidative stress was investigated using LLC-PK(1) renal tubular epithelial cells, which are susceptible to oxidative stress. MG (500 microM) treatment induced LLC-PK(1) cell death to nearly 50% compared with non-treated control cells, but lipoic acid significantly inhibited the MG-induced cytotoxicity in a concentration-dependent manner. In addition, lipoic acid treatment dose-dependently reduced the intracellular reactive oxygen species level increased by 500 microM MG. The nitric oxide level was also increased by 500 microM MG treatment, but it was significantly inhibited by lipoic acid. Furthermore, lipoic acid treatment at 50 microM inhibited the nuclear translocation of nuclear factor-kappa B induced by MG treatment in LLC-PK(1) cells. These findings indicate that lipoic acid has potential as a therapeutic agent against the development of diabetic complications related to MG-induced oxidative stress in diabetes. PMID- 18388416 TI - Tanning can be an alternative source of vitamin d in high risk populations. PMID- 18388417 TI - Prognosis of canine patients with nasal tumors according to modified clinical stages based on computed tomography: a retrospective study. AB - To evaluate the efficacy of clinical staging based on computed tomography (CT) imaging over the World Health Organization (WHO) staging system based on radiography for nasal tumors in dogs, a retrospective study was conducted. This study used 112 dogs that had nasal tumors; they had undergone radiography and CT and had been histologically confirmed as having nasal tumors. Among 112 dogs, 85 (75.9%) were diagnosed as adenocarcinoma. Then they were analyzed for survival time according to each staging system. More than 70% of the patients with adenocarcinoma were classified as having WHO stage III. The patients classified under WHO stage II tended to survive longer than those classified under WHO stage III. Dogs classified under WHO stage III were further grouped into CT stages III and IV, and CT stage III patients had a significantly longer survival time than CT stage IV patients. In addition, patients treated with a combination of surgery and radiation had a significantly longer survival time than the patients who did not receive any treatment in CT stage III. On the other hand, different treatment modalities did not show a significant difference in the survival time of CT stage IV dogs. The results suggest that WHO stage III dogs may have various levels of tumor progression, indicating that the CT staging system may be more accurate than the WHO staging system. PMID- 18388418 TI - Removal of 65Zn from mouse body by isotopic dilution and by DTPA chelation. AB - Isotopic dilution with stable zinc and chelation with DTPA are recommended for removal of radioactive zinc from the body; however, it is unclear which method is more effective. In the present study, the efficacies of these methods were compared in order to determine which treatment should be selected in case of internal contamination with radioactive zinc. Intraperitoneal administration of stable zinc dose-dependently removed 65Zn from the mouse body. However, the dose could not be increased above 3 mg/kg due to its toxicity. Oral administration of zinc was less effective than intraperitoneal administration at the same dose. Our results suggest that the recommended dose of stable zinc (2-3 mg/kg, p.o.) has little efficacy. The efficacies of Ca-DTPA and Zn-DTPA were strongly dependent on the elapsed time after 65Zn exposure. Zn-DTPA was more effective than Ca-DTPA, and its recommended dose (30 micromol/kg) significantly removed 65Zn. Therefore, chelation therapy with Zn-DTPA should be started as soon as possible after internal contamination with radiozinc. PMID- 18388419 TI - A macroscopic examination of M. biceps femoris and M. gluteus maximus in the orangutan. AB - The musculature of the hip and thigh in the orangutan has been described previously. Anatomically, there are various descriptions among primates in those structures, in particular, the relationship between M. biceps femoris and M. gluteus maximus, their derivatives, and the muscle segment. However, a detailed innervation system to this ischiofemoral part has not been described, thus there is still uncertainty as to with which muscle it is associated. In this analysis, we examined the gross anatomy of the hip and thigh muscles of the orangutan and chimpanzee, including their innervation. Also, a comparison was made with documented data of other primates. As a result of these observations, it was found that the ischiofemoral part in the orangutan is innervated by the same sciatic nerve branch (the common peroneal nerve) as the long head of M. biceps femoris, but not by the same nerve as M. gluteus maximus. Therefore, the ischiofemoral part is appropriately considered as a part of the long head of M. biceps femoris. It appears that this morphologic feature is an adjustment to the arboreal life of the orangutan. The development of the flexor complex of the thigh is necessary for this arboreal adaptation, resulting in a unique musculature of M. biceps femoris in the orangutan. PMID- 18388420 TI - Antibiotic furazolidone induces CYP1A but not CYP2E1 subfamily in rat liver. AB - Furazolidone (FZ), one of the nitrofuran fungicides, is used as a veterinary medicine in the Middle and Far Eastern countries. In this study, FZ (125 mg/kg) was administered orally to Wistar rats for 3 days. Results of the Ames test using the S-9 fraction of rats treated with FZ showed a significant increase in the number of revertant colonies. Western blot analysis of hepatic CYP isozymes induced by FZ, revealed a remarkable induction of CYP1A1 apoprotein, but CYP1A2 and CYP2E1 apoproteins were not altered. In addition, the expression of CYP1A1 mRNA level in rats treated with FZ by RT-PCR was significantly enhanced by FZ treatment. We concluded that FZ is apparently mutagenic and induces transcription of the CYP1A1 isozyme, which metabolically activates numerous promutagens, in hepatocytes. PMID- 18388421 TI - Vitellogenin, a biomarker for environmental estrogenic pollution, of Reeves' pond turtles: analysis of similarity for its amino acid sequence and cognate mRNA expression after exposure to estrogen. AB - Vitellogenin (VTG), a biomarker for environmental estrogenic pollution, can be detected in the bloodstream of oviparous animals before morphological and functional abnormalities appear due to exposure to environmental estrogens. Reports observing VTG in turtles have been limited. We therefore cloned and sequenced a partial cDNA of VTG in Reeves' pond turtle, Chinemys reevesii. The cloned cDNA fragment possessed the start codon and 2,229 bp, encoding 743 amino acid residues. A sequence of deduced amino acid from the cDNA did not contain a high serine content, such as that which exists in phosvitin. Two N-glycosylation sites were found in the sequence. The sequence was compared to those of two birds (chicken and herring gull), one amphibian (Xenopus), and five fishes (carp, zebrafish, eel, haddock, and red seabream). The C. reevesii VTG was similar to that of herring gull (78%, value of positives), chicken (76%), Xenopus (69%), eel (63%), red seabream (62%), haddock (62%), carp (62%), and zebrafish (61%). The phylogenetic tree showed that C. reevesii VTG existed between the amphibian and birds, and it was present far from fish VTGs. A reverse transcription-polymerase chain reaction method was employed to detect the mRNA expression of the C. reevesii VTG through the use of primers designed from our sequence. The VTG mRNA expression (292 bp) was proven in the total RNA extraction from the liver of the juvenile turtles which were treated with estradiol-17beta. The information herein would be useful for ecotoxicological studies using freshwater turtles and these findings are expected to contribute positively towards wildlife conservation. PMID- 18388422 TI - Ultrastructural study on the epithelial responses against attachment of indigenous bacteria to epithelial membranes in peyer's patches of rat small intestine. AB - The ultrastructure of epithelial responses against the membrane adhesion of indigenous bacteria was investigated in the follicle-associated epithelium (FAE) of rat small intestine. The most frequent adherence of the various morphological types of bacteria to the epithelial membranes was found at the apex of the FAE. The attachment sites were deeply invaginated, and their bottoms were deformed into a sharp cone shape. Four layers with different electron densities were formed just beneath the apical membranes by microfilaments which surrounded the invaginations. The electron density of each layer was gradually decreased as being apart from the invaginations. The extremities of some bacteria in the invaginations were deformed into sharpened shapes. The cell walls of the extremities of the bacteria were occasionally dissolved in the invaginations, and their cytoplasms were slightly swollen with low electron densities. In some invaginations, the attached bacteria were eliminated to leave their fragments such as filamentous debris and a part of cell walls. Finally these remnants disappeared completely. When the bacterial colonies existed in the middle region of the FAE, the attachment of bacteria resulted in the engulfment of bacteria by M cells. The degenerated bacteria whose cytoplasmic matrices were separated into high electron dense materials and cleared materials were occasionally engulfed by ordinary microvillous columnar epithelial cells or goblet cells throughout the FAE. These findings suggest that the epithelial cells reject the attachment of live indigenous bacteria and that the M cells absorb indigenous bacteria in rat Peyer's patches. PMID- 18388423 TI - Efficacy of intravaginal progesterone administration as an additional treatment on two types of timed AI protocols in a commercial herd of Holstein heifers. AB - Reproductive performance of two types of timed artificial insemination (TAI) protocols with or without intravaginal progesterone insert (CIDR) was investigated in a commercial herd of Holstein heifers. A total of seventy-four heifers with 14.4 months of age were allocated to two groups; Ovsynch (n=44) and estradiol benzoate (EB) used Heatsynch (EB-Heatsynch, n=30), and each group was additionally divided into two subgroups with CIDR insertion from day 0 to 7 (n=36) and without CIDR group (n=38). Blood was collected for progesterone (P4) analysis and ovarian finding was monitored with ultrasonography. Heifers in CIDR treated group resulted in higher pregnancy rate as compared to No-CIDR-treated group (63.9% vs 21.1%, P<0.01). Heifers with functional corpus luteum (CL) on day 0 resulted in significantly higher pregnancy rate in CIDR-treated group than No CIDR-treated group (day 0: 67.9% vs 13.0%, P<0.01). CIDR insertion suppressed the intermediate ovulation during the first 7 days and the period from the second GnRH or EB administration to TAI as compared to No-CIDR-treated group (first 7 days: 33.3% vs. 52.6%; P<0.05, before TAI: 11.1% vs. 37.0%; P<0.05). In conclusion, the selected TAI protocols with CIDR provided acceptable pregnancy rate and contributed to the economical improvement by shortening the average age of first calving approximately for 2.5 months as compared to the previous management without TAI protocols. PMID- 18388424 TI - Distribution of the lingual tonsils of cattle designated as specified risk materials. AB - The tonsils of cattle, including palatine tonsils, pharyngeal tonsils, tubal tonsils and lingual tonsils, are designated as specified risk materials (SRM). However, the detailed distribution of lingual tonsils in cattle is unknown. We therefore histologically examined their distribution in 198 tongue specimens from cattle. The examinations confirmed that the presence of lingual tonsils was limited to the tissue of the lamina propria on the dorsal and lateral aspects of the tongue, not reaching the muscular layer below. More than 90% of the lingual tonsils were located between the distribution center of the vallate papillae and the radix linguae (root of the tongue). However, they were also found in the area extending from the lingual torus to the rostral-most vallate papilla in an individual, suggesting that the complete removal of the lingual tonsils requires elimination of the lamina propria extending from the lingual torus to the radix linguae. PMID- 18388425 TI - Mechanisms responsible for reduced cardiotoxicity of mitoxantrone compared to doxorubicin examined in isolated guinea-pig heart preparations. AB - We reported previously that doxorubicin, an anticancer agent that has an anthracycline structure, alters Ca2+ releasing and uptake mechanisms in the sarcoplasmic reticulum of myocardial cells. These effects of doxorubicin are apparently related to its cardiotoxicity. Mitoxantrone is a similar anticancer agent with an anthracenedion structure that has been shown to be significantly less cardiotoxic. In the present study, the effects of mitoxantrone on the functions of the sarcoplasmic reticulum were examined in isolated muscle preparations obtained from the guinea-pig heart. In electrically-stimulated left atrial muscle preparations, incubation in vitro for 4 hr with 30 or 100 microM mitoxantrone significantly prolonged the time to the peak of twitch tension, markedly increased the developed tension observed at lower stimulation frequencies, thereby attenuating the slope of positive force-frequency relationships, and increased the postrest contraction observed after a 60-sec quiescent period. In myocytes isolated from ventricular muscles, 30 microM mitoxantrone increased the peak and the size of intracellular Ca2+ concentrations ([Ca2+] i), and prolonged the time to peak [Ca2+]i. In skinned muscle fiber preparations obtained from the left ventricular muscle, 30 muM mitoxantrone significantly increased the caffeine-induced contraction without affecting the Ca2+ sensitivity of contractile proteins. These results suggest that mitoxantrone enhances Ca2+ release from the sarcoplasmic reticulum in isolated atrial muscle preparations obtained from the guinea-pig heart. Apparent enhancement of the sarcoplasmic reticulum functions, in contrast to anthracyclines that has been shown to suppress these functions, seems to explain the relative lack of marked cardiotoxicity of mitoxantrone. PMID- 18388426 TI - Immunohistochemical distribution of inwardly rectifying K+ channels in the medulla oblongata of the rat. AB - The inwardly rectifying K+ channels, Kir1.1, Kir2.3 and Kir4.1-Kir5.1, are the candidate chemosensory molecules for CO2/H+. We determined the mRNA expression and immunohistochemical localization of these channels in the medulla oblongata of the rat. RT-PCR analysis revealed mRNAs of Kir1.1, Kir2.3, Kir4.1 and Kir5.1 were detected in the medulla. The immunoreactivities for Kir1.1, Kir2.3, Kir4.1, and Kir5.1 were observed in the medulla, and immunolabeling pattern was varied by the subunit. Immunoreactivities for Kir1.1 and Kir2.3 were observed in the nerve cell bodies and glial cells both in the chemosensory areas [nucleus tractus solitarius (NTS), nucleus raphe obscurus (RO), pre-Botzinger complex (PreBotC)] and non-chemosensory area [hypoglossal nucleus (XII), inferior olive nucleus (IO)]. Kir4.1 immunoreactivity was observed in the glial cells and neuropil, especially in XII and IO. Kir5.1 immunoreactivity was observed in the nerve cell bodies in the XII, RO, and PreBotC, but not in the NTS or IO. In the NTS, a dense network of varicose nerve fibers showed immunoreactivity for Kir5.1. Our findings suggest that Kir channels may not act specific to the central chemoreception, but regulate the ionic properties of cellular membranes in various neurons and glial cells. PMID- 18388427 TI - Differences in lipid peroxidation and Cu,Zn-superoxide dismutase in the hippocampal CA1 region between adult and aged dogs. AB - Reactive oxygen species have been long associated with oxidative stress relevant to many pathological damages. In brain, 4-hydroxy-2E-nonenal (HNE), a major cytotoxic end product of lipid peroxidation, is produced. In contrast, superoxide dismutase (SOD), one of the major antioxidant enzymes, protects neurons from oxidative stress. The aim of this study is to observe differences in the distribution of HNE and Cu,Zn-superoxide dismutase (SOD1) in the hippocampal CA1 region of adult (2-3 years of age) and aged (10-12 years of age) dogs. The HNE immunoreactivity and protein level in the CA1 region were significantly high in the aged dogs compared to those in the adult dogs. SOD1 immunoreactivity and its protein level were also higher in the aged dogs than those in the adult dogs. However, there were not significant differences in NeuN (a neuron-specific soluble nuclear antigen) immunoreactivity in CA1 neurons between the adult and aged dogs. These differences may be associated with oxidative stress in aged dogs compared to that in adult dogs. PMID- 18388428 TI - Long-term clinical evaluation of mitral valve replacement with porcine bioprosthetic valves in dogs. AB - This study evaluated the long-term clinical performance of newly developed porcine bioprosthetic valves cross-linked with glutaraldehyde and polyepoxy compound for mitral valve replacement (MVR) in dogs. Five beagle dogs underwent MVR using the porcine bioprosthetic valves during cardiopulmonary bypass. Antithrombotic drugs were administered only for one month after MVR. Six months after MVR, transvalvular regurgitation was not observed in all dogs, paravalvular leakage was seen only in one dog. Twelve months after MVR, mild transvalvular regurgitations were observed in two dogs. Although diastolic atrioventricular pressure gradient was increased gradually, no significant differences were observed. Pressure half-time and valve area were within normal ranges as the bioprosthetic value. There was no clinical symptom of the thrombosis and the thrombogenesis was not observed in the porcine bioprosthetic valve and the annulus in all dogs for twelve months after MVR. The clinical findings suggest that antithrombogenicity of the valves were maintained, though the duability might not be enough in the long-term period. PMID- 18388429 TI - Flow cytometric analysis of canine umbilical cord blood lymphocytes. AB - Lymphocyte subsets in canine umbilical cord blood were flow cytometrically analyzed and compared with those of the dams' peripheral blood. The proportion of CD3+ T lymphocytes, CD21+CD3- B lymphocytes, and CD3-CD21- non-T non-B lymphocytes in umbilical cord blood was 52.9%, 30.4%, and 16.7%, respectively. T lymphocyte/B lymphocyte ratio was significantly lower in the umbilical cord blood than in the dams' peripheral blood (2.1 +/- 1.4 versus 11.0 +/- 8.1, P < 0.001). In contrast, CD4+ lymphocyte/CD8+ lymphocyte ratio was significantly higher in the umbilical cord blood than in the dams' peripheral blood (7.6 +/- 2.2 versus 1.8 +/- 0.6, P<0.001). These findings clarified the phenotypic characters of canine umbilical cord blood lymphocytes. PMID- 18388430 TI - Polypoid eosinophilic cystitis with pseudosarcomatous proliferative tissue in a dog. AB - A dog presented with hematuria, and two small polypoid masses were detected in the urinary bladder. Histopathologically, the masses were located in the mucosal or submucosal layer. That tissue consisted of a random proliferation of spindle shaped, round and pleomorphic cells with single or multiple large atypical nuclei and abundant cytoplasm, and eosinophil infiltration. These large cells were confirmed by immunohistochemical staining as fibroblasts, myofibroblasts and macrophages. Mitotic figure was rarely seen. These masses were diagnosed as eosinophilic polypoid cystitis with pseudosarcomatous proliferative tissue, since they consisted of a wide variety of cells and showed low growth activity. PMID- 18388431 TI - Clinical and pathological aspects of hemophilia a in Japanese Brown cattle. AB - A coagulopathy with subcutaneous bleeding and muscular or peritracheal/periesophageal bleeding occurred in two male Japanese Brown calves of the same dam. One of the affected calves died three days after the onset of bleeding and the other survived normally until being slaughtered despite once suffering from subcutaneous hematoma. Hemostatic tests of the latter case showed prolonged activated partial thromboplastin time (APTT), and severely reduced factor VIII activity. In addition, von Willebrand factor activity, determined by the human platelet aggregation test, was within the normal range; therefore, the calf was diagnosed with hemophilia A. These are the first bovine cases of hemophilia A definitely diagnosed clinicopathologically. PMID- 18388433 TI - Histopathological comparison of pulmonary artery lesions between raccoon dogs (Nyctereutes procyonoides) and domestic dogs experimentally infected with Dirofilaria immitis. AB - Five raccoon dogs (Nyctereutes procyonoides) and two domestic dogs (Canis familiaris) were subcutaneously infected with 100 infective larvae (L3) of Dirofilaria immitis. Two and five worms, respectively, were collected from two of three raccoon dogs. Villous endarteritis was found in the raccoon dog with five worms and two dogs at 116 days after infection. The number of recovered worms in the raccoon dogs was significantly smaller than that of the domestic dogs having 22 and 29 worms, while histopathological features and the severity of the lesions in the raccoon dogs were similar to those in the domestic dogs. The vascular lesions in two chronically-infected raccoon dogs turned into much severe at 565 and 590 days after inoculation. PMID- 18388432 TI - Pedigree analysis of factor XI deficiency in Japanese black cattle. AB - Using a DNA-based diagnostic test for factor XI deficiency in Japanese black cattle, we surveyed 123 cattle (42 sires and 81 dams) in Gifu and Hyogo prefectures, and calculated gene frequencies. In sires, we drew up the pedigree network of the cattle with the factor XI deficiency. Results showed that the mutated allele of factor XI deficiency was retroactive in at least 6 or more generations of sires. Frequencies of the mutant gene were higher at 26.4% in total, and at 33.3% in sires. All 7 cattle with the homozygote of mutated allele were clinically normal, and showed no bleeding episodes. The mutated allele of factor XI deficiency might be widespread among Japanese black cattle. PMID- 18388434 TI - Immunohistochemical distribution of viral antigens in pigs naturally infected with porcine teschovirus. AB - A distribution of porcine teschovirus (PTV) antigens in pigs naturally infected with PTV is presented using the method of immunohistochemical examination. In the nervous system, PTV antigens were found in the cytoplasm of neuronal cells and glial cells distributed in the spinal ventral horn and brain stem, and also in the cytoplasm of ganglion cells in the spinal ganglion. No antigens were seen in the cerebral hemisphere. In the nervous system, the distribution of PTV antigens was consistent with lesions characteristic of nonsuppurative encephalomyelitis. In the other examined organ, PTV antigens were observed in bronchiolar epithelial cells in the lung, hepatocytes in the liver, epithelial cells in the tonsils and the myenteric nerve plexus in the small and large intestine. PMID- 18388435 TI - Relative quantification of canine CD56 mRNA expression by real-time polymerase chain reaction method in normal tissues and activated lymphocytes. AB - Real-time PCR was optimized for the quantification of canine CD56 mRNA expression. This study was conducted to easily quantify canine CD56 expression and to identify its expression in normal tissues, peripheral blood mononuclear cells and activated lymphocytes in dogs. This assay revealed the highest level of CD56 mRNA expression in the normal canine brain, followed by the lung, kidney and liver. CD56 mRNA expression level in peripheral blood mononuclear cells was considerably lower; among activated lymphocytes in vitro, CD56 mRNA expression was increased. PMID- 18388436 TI - Trans-endothelial and trans-epithelial transfer of lactoferrin into the brain through BBB and BCSFB in adult rats. AB - The transportation of intravenously administered bovine lactoferrin (bLF) into the cerebrospinal fluid (CSF) was immunohistochemically investigated in adult rats. Administered bLF was detected in the vesicular membranes of endothelial cells in cerebral blood vessels 10 min after the infusion. Numerous immunoreactive small vesicles were also detected at the ependymal cells in the choroid plexus. Moreover, the bLF concentration in the CSF was significantly increased at 1-2 hr after the intravenous infusion of bLF (10 or 30 mg/kg). These findings clearly demonstrate that LF is possibly transported into the brain matter even in adult animals. PMID- 18388437 TI - Aldosterone-, corticosterone- and cortisol-secreting adrenocortical carcinoma in a dog: case report. AB - A 12-year-old, intact female beagle exhibited symptoms of polyuria-polydipsia and hyperorexia for two months. Blood tests showed elevated asparate aminotransferase, alanine aminotransferase, alkaline phosphatase and creatine kinase levels, as well as marked hypokalemia. The results of adrenocorticotropic hormone stimulation test showed elevated cortisol, aldosterone and corticosterone concentrations. Abdominal ultrasonography confirmed a mass in the left adrenal gland. Masses were also seen in the liver and caudal vena cava. Diagnosis was a tumor of the adrenal cortex with metastases. Trilostane administration was initiated. The dog initially showed improved demeanor as a result of regulating hormone secretion. However, after 88 days, the dog weakened rapidly, before dying on the 117th day. Pathological findings confirmed a diagnosis of adrenocortical carcinoma. PMID- 18388438 TI - Effects of active egg white product/ Clostridium butyricum Miyairi 588 additive on peripheral leukocyte populations in periparturient dairy cows. AB - The leukocyte populations of periparturient dairy cows were analyzed after administration of active egg white/Clostridium butyricum Miyairi additive. Sixty eight Holstein milking cows were divided into 3 groups. Group A was administered active egg white product (AEWP)/Clostridium butyricum Miyairi 588 (Miyairi 588) additive (n=23). Group B was administered Miyairi 588 only (n=23), and Group C was the control group (n=22). The challenged groups were administered 100 g of AEWP + Miyairi 588, or Miyairi 588 alone, daily for 60 days from 1 month before until 1 month after paturition. Blood samples were collected from all groups three times (1 month before, 1 week after and 1 month after parturition) for analysis of the peripheral leukocyte population. The results showed significantly higher numbers of CD4+ cells in Group A compared with Group C 1 week after paturition. AEWP/Miyairi 588 additive may enhance the number of CD4+ T cells in periparturient dairy cows. PMID- 18388439 TI - [Development from autoimmune pancreatitis to systemic disease]. PMID- 18388440 TI - [Diagnosis of autoimmune pancreatitis with special reference to comparison of diagnostic criteria among Japan, Korea and United States]. PMID- 18388441 TI - [Immunological aspects of autoimmune pancreatitis]. PMID- 18388442 TI - [Characteristic findings of sclerosing cholangitis with autoimmune pancreatitis]. PMID- 18388443 TI - [New development about autoimmune pancreatitis]. PMID- 18388444 TI - [A case of gastric carcinoid with type A gastritis]. AB - A 40-year-old man was admitted to our hospital for treatment of gastric carcinoid of 7-mm in diameter associated with type A gastritis. Endoscopic mucosal resection was performed and histological examination revealed a gastric carcinoid with submucosal invasion (2500microm). Three months after the resection, the other carcinoid of 3-mm lesion was found, but no recurrence of resected portion. Neither local recurrence nor distant metastasis has been observed during 1-year follow-up period. These findings revealed that gastric carcinoid with type A gastritis, when smaller than 1-cm in diameter, could be cured with endoscopic procedure. PMID- 18388445 TI - [A case of gastrointestinal AL amyloisosis requiring emergency surgery]. AB - A 69-year-old man tarry stools received emergency endoscopy. Which revealed a solitary submucosal tumor about 15mm in diameter with a central ulcer, exhibiting woozing bleeding in the duodenal second portion. Endoscopic hemostasis was unsuccessful so emergency surgery was performed. Histological examination revealed amyloid with A-lambda immunoreactivity. Usually, this type of amyloidosis appears multiple submucosal masses. However this case presented as a single nodule at initial examination. We should keep in mind the potential of encountering this atypical form of amyloidosis. PMID- 18388446 TI - [A case of polypoid type ganglioneuroma of the decsending colon]. AB - A 32-year-old woman came to our hospital with purpose of careful examination for anemia. Colonoscopy was revealed a solitary protrusion with irregular shape covered with red and discolored mucosa of the descending colon. Surface of this lesion was smooth, which had flexibility in dynamic study with infusion of the air and water. Additionally, multiple ulcer scars was recognized on this lesion. After dye-spraying using by crystal-violet, no neoplastic pits was detected on the lesion except for asteroidal pits and pattern of pinecone on reddish protrusions. Endoscopic ultrasonography demonstrated a thickened low echoic layer (from 2nd to upper half of the 3rd layer) and anechoic structure in higher reddish part of the lesion. Diagnostic EMR was performed with no complication. Histological examination was revealed a spindle cells and ganglion cells in much fiber which was positive for immunostaining of S-100 ptotein. Hyperplastic glands were seen with no neoplastic change in reddish protrusion. Therefore, Diagnosis of polypoid ganglioneuroma of the descending colon was made. To a rare thing, our case was the 16th reported case of ganglioneuroma in large intestine without neurofibroma-1 or multiple endocrine neoplasm. PMID- 18388447 TI - [A case of focal nodular hyperplasia presenting corona enhancement on single level dynamic CT during hepatic arteriography]. AB - A hepatic nodule was detected in segment 5/6 on abdominal US study in a 28 year old male. The nodule was 7cm in diameter and the early phase of contrasted US, CT and MRI images showed spoke-wheel like vessels radiating from the center. No defect images were observed on postvascular phase contrasted US and SPIO MRI, which indicated the presence of Kupffer cells in the nodule. The nodule was diagnosed as a focal nodular hyperplasia (FNH) based on histological findings. The late phase of single level dynamic CT during hepatic arteriography (CTHA) showed corona enhancement of the nodule, which is considered to be characteristic of hypervascular metastatic liver tumors, hyperplastic nodules and HCCs. In our case, the drainage flow from the nodule may have been visualized as corona enhancement via the pathway from the sinusoid in the nodular periphery to the one in the adjacent and contiguous parenchyma. PMID- 18388448 TI - [A case of double common bile duct with gastric ulcer]. AB - A 51-year-old man visited the Sasebo General Hospital because of a niche and a ductal lesion with reflux of barium in the lesser curvature of the gastric body in the upper gastrointestinal series of an annual medical checkup. Endoscopic examination showed an ulcer and a depressed lesion draining yellowish serous liquid in the stomach. Abdominal CT scan and MRCP revealed the ductal structure reached from the lesser curvature of the gastric body to the left lobe of the liver besides the ordinary pancreatobiliary system. ERCP showed normal common bile duct, gall bladder, and right intrahepatic bile duct, while the fistulography using cannula through the gastric depressed lesion revealed an accessory bile duct communicating to the left intrahepatic bile duct. The both biliary systems were completely separated, resulting in the diagnosis of double common bile duct of type IIIa. PMID- 18388449 TI - [A case report of anaphylactoid purpura with acute abdominal pain secondary to trauma caused by traffic accident]. AB - A 17-year-old man was admitted to our hospital with multiple fractures resulting from traffic accident. After treatment of fractures, his general status was improved. However, one month after traffic accident, he suffered severe pain in the epigastrium. Ultrasonography and computed tomography showed thickening of the intestinal wall in the duodenum, ileum, and ascending colon. Nineteen days after the onset of abdominal pain, small hemorrhagic spots appeared on both of the lower legs. Subsequently, he developed proteinuria and hematuria. Purpura nephritis was diagnosed in biopsy specimens of the kidney. Anaphylactoid purpura associated with traffic accident is very rare and it is difficult to diagnose without skin and renal symptoms. PMID- 18388450 TI - [A case of splenic inflammatory pseudotumor]. AB - A 51-year-old woman who had undergone gastrectomy for advanced gastric cancer was found to have a splenic tumor during the postoperative clinical observation. Abdominal computed tomography (CT) demonstrated solitary splenic tumor 15mm in diameter with delayed contrast enhancement. Abdominal ultrasonography (US) revealed low echoic mass with enhancement at vascular and perfusion image. We performed splenectomy to exclude the possibility of the metastatic tumor. The tumor was histopathologically diagnosed as inflammatory pseudotumor because of the presence of acidophilic fiber proliferation, hyalinized tissue and infiltration of lymphocytes and plasma cells. PMID- 18388451 TI - [A case of jejunal perforation in gallstone ileus]. AB - Gallstone ileus is a rare but important cause of small bowel obstruction in the geriatric population. A 65-year-old man with a twenty year history of cholecystolithiasis was admitted to our hospital with abdominal pain and vomiting. Physical exams showed abdominal defence and rebound tenderness. A plain abdominal X-ray suggested a small bowel obstruction and pneumobilia. CT scan revealed a 2.5-cm gallstone at the jejunum and air in the biliary tree. The patient underwent a emergency laparotomy based on a diagnosis of panperitonitis with a perforation associated with gallstone ileus. Operative findings revealed a jejunal perforation and a impacted stone on the anal side of perforation. Enterolithotomy and jejunal resection were performed with cholecystectomy and repairment of the cholecystoduodenal fistula. PMID- 18388452 TI - [Effects of nanoparticles on humans]. AB - Methods and end-points used in assessing the hazards of fibrous materials, whose hazards have been assessed, are often employed in studies on the hazards of nanoparticles. Since a possible route of exposure to nanoparticles in work environments is inhalation, data from animal inhalation and intratracheal instillation studies are thought to provide useful insights for assessing the hazards of such particles. Persistent inflammation and fibrosis in the chronic phase and their related factors are particularly important indicators of lung injury. On the other hand, there are some differences in the results of hazard assessment studies due mainly to a lack of detailed physiochemical characterizations of the nanoparticles evaluated. Dispersion properties in the chambers just before exposure in inhalation studies and those in the suspension to be injected in intratracheal instillation studies need to be determined. The number of hazard assessment studies in which characterization of nanoparticles is conducted, while small at present, is increasing and this could consequently yield reliable hazard assessment and risk assessment. PMID- 18388453 TI - [Perceived health and its correlation with depression and subjective symptoms of nurses and care workers]. AB - The objective of this study was to identify psychological and physical health characteristics of nurses and care workers employed at a psychiatric hospital and to characterize depressive symptoms by means of correlation analysis. Subjects were 89 nurses and 78 care workers employed at a psychiatric and rehabilitation hospital in Gunma Prefecture, Japan. They responded to a structured questionnaire, the Total Health Index (THI). Mean scores of the four THI scales, Vague Complaints, Respiratory, Depression and Neurotics, of all the 167 study subjects were significantly higher than those of three reference groups. The smoking rate was almost two times higher than that of adult Japanese women. These results may be attributed to psychological stress these workers suffer in their daily human service. However, we found little difference in THI scores based on shift work or responsibilities (nursing vs. care taking). Our findings suggest that it is necessary to provide these workers some support for stress management. THI scores that correlated most strongly with the Depression score were Mental Instability, Irregularity of Life, Irritability, and Vague Complaints (correlation coefficients were 0.75, 0.54, 0.53, and 0.52, respectively). THI scores that correlated most weakly with the Depression score were Digestive, Respiratory, and Mouth and Anal (correlation coefficients were 0.35, 0.34, and 0.32, respectively). Depression in our subjects appeared to be related exclusively to vague psychological symptoms rather than organ-specific symptoms. PMID- 18388454 TI - sanN encoding a dehydrogenase is essential for Nikkomycin biosynthesis in Streptomyces ansochromogenes. AB - Nikkomycins are a group of peptidyl nucleoside antibiotics with potent fungicidal, insecticidal, and acaricidal activities. sanN was cloned from the partial genomic library of Streptomyces ansochromogenes 7100. Gene disruption and complementation analysis demonstrated that sanN is essential for nikkomycin biosynthesis in S. ansochromogenes. Primer extension assay indicated that sanN is transcribed from two promoters (sanN-P1 and sanN-P2), and sanN-P2 plays a more important role in nikkomycin biosynthesis. Purified recombinant SanN acts as a dehydrogenase to convert benzoate-CoA to benzaldehyde in a random-order mechanism in vitro, with respective Kcat/Km values of 3.8 mM-1s-1 and 12.0 mM-1s-1 toward benzoate-CoA and NADH, suggesting that SanN catalyzes the formation of picolinaldehyde during biosynthesis of nikkomycin X and Z components in the wild type stain. These data would facilitate us to understand the biosynthetic pathway of nikkomycins and to consider the combinatorial synthesis of novel antibiotic derivatives. PMID- 18388455 TI - Functional analysis of a gene encoding endoglucanase that belongs to glycosyl hydrolase family 12 from the brown-rot basidiomycete Fomitopsis palustris. AB - The brown-rot basidiomycete Fomitopsis palustris is known to degrade crystalline cellulose (Avicel) and produce three major cellulases, exoglucanases, endoglucanases, and beta- glucosidases. A gene encoding endoglucanase, designated as cel12, was cloned from total RNA prepared from F. palustris grown at the expense of Avicel. The gene encoding Cel12 has an open reading frame of 732 bp, encoding a putative protein of 244 amino acid residues with a putative signal peptide residing at the first 18 amino acid residues of the N-terminus of the protein. Sequence analysis of Cel12 identified three consensus regions, which are highly conserved among fungal cellulases belonging to GH family 12. However, a cellulose-binding domain was not found in Cel12, like other GH family 12 fungal cellulases. Northern blot analysis showed a dramatic increase of cel12 mRNA levels in F. palustris cells cultivated on Avicel from the early to late stages of growth and the maintenance of a high level of expression in the late stage, suggesting that Cel12 takes a significant part in endoglucanase activity throughout the growth of F. palustris. Adventitious expression of cel12 in the yeast Pichia pastoris successfully produced the recombinant protein that exhibited endoglucanase activity with carboxymethyl cellulose, but not with crystalline cellulose, suggesting that the enzyme is not a processive endoglucanase unlike two other endoglucanases previously identified in F. palustris. PMID- 18388456 TI - Cloning, expression, and characterization of protease-resistant xylanase from Streptomyces fradiae var. k11. AB - The gene SfXyn10, which encodes a protease-resistant xylanase, was isolated using colony PCR screening from a genomic library of a feather-degrading bacterial strain Streptomyces fradiae var. k11. The full-length gene consists of 1,437 bp and encodes 479 amino acids, which includes 41 residues of a putative signal peptide at its N terminus. The amino acid sequence shares the highest similarity (80%) to the endo-1,4-beta-xylanase from Streptomyces coelicolor A3, which belongs to the glycoside hydrolase family 10. The gene fragment encoding the mature xylanase was expressed in Escherichia coli BL21 (DE3). The recombinant protein was purified to homogeneity by acetone precipitation and anion-exchange chromatography, and subsequently characterized. The optimal pH and temperature for the purified recombinant enzyme were 7.8 and 60 degrees , respectively. The enzyme showed stability over a pH range of 4-10. The kinetic values on oat spelt xylan and birchwood xylan substrates were also determined. The enzyme activity was enhanced by Fe2+ and strongly inhibited by Hg2+ and SDS. The enzyme also showed resistance to neutral and alkaline proteases. Therefore, these characteristics suggest that SfXyn10 could be an important candidate for protease resistant mechanistic research and has potential applications in the food industry, cotton scouring, and improving animal nutrition. PMID- 18388457 TI - Functional effects of increased copy number of the gene encoding proclavaminate amidino hydrolase on clavulanic acid production in Streptomyces clavuligerus ATCC 27064. AB - The effect of increasing levels of proclavaminate amidino hydrolase (Pah) on the rate of clavulanic acid production in Streptomyces clavuligerus ATCC 27064 was evaluated by knock-in a gene (pah2) encoding Pah. A strain (SMF5703) harboring a multicopy plasmid containing the pah2 gene showed significantly retarded cell growth and reduced clavulanic acid production, possibly attributable to the deleterious effects of the multicopy plasmid. In contrast, a strain (SMF5704) carrying a single additional copy of pah2 introduced into chromosome via an integrative plasmid showed enhanced production of clavulanic acid and increased levels of pah2 transcripts. Analysis of transcripts of other genes involved in the clavulanic acid biosynthetic pathway revealed a pattern similar to that seen in the parent. From these results, it appears that clavulanic acid production can be enhanced by duplication of pah2 through integration of a second copy of the gene into chromosome. However, increasing the copy number of only one gene, such as pah2, does not affect the expression of other pathway genes, and so only modest improvements in clavulanic acid production can be expected. Flux controlled by Pah did increase when the copy number of pah2 was doubled, suggesting that under these growth conditions, Pah levels may be a limiting factor regulating the rate of clavulanic acid biosynthesis in S. clavuligerus. PMID- 18388458 TI - Expression and characterization of polyketide synthase module involved in the late step of cephabacin biosynthesis from Lysobacter lactamgenus. AB - The cephabacins produced by Lysobacter lactamgenus are beta-lactam antibiotics composed of a cephem nucleus, an acetate residue, and an oligopeptide side chain. In order to understand the precise implication of the polyketide synthase (PKS) module in the biosynthesis of cephabacin, the genes for its core domains, beta ketoacyl synthase (KS), acyltransferase (AT), and acyl carrier protein (ACP), were amplified and cloned into the pET-32b(+) expression vector. The sfp gene encoding a protein that can modify apo-ACP to its active holo-form was also amplified. The recombinant KS, AT, apo-ACP, and Sfp overproduced in the form of His6-tagged fusion proteins in E. coli BL21(DE3) were purified by nickel-affinity chromatography. Formation of stable peptidyl-S-KS was observed by in vitro acylation of the KS domain with the substrate [L-Ala-L-Ala-LAla- L-3H-Arg] tetrapeptide-S-N-acetylcysteamine, which is the evidence for the selective recognition of tetrapeptide produced by nonribosomal peptide synthetase (NRPS) in the NRPS/ PKS hybrid. In order to confirm whether malonyl CoA is the extender unit for acetylation of the peptidyl moiety, the AT domain, ACP domain, and Sfp protein were treated with 14C-malonyl-CoA. The results clearly show that the AT domain is able to recognize the extender unit and decarboxylatively acetylated for the elongation of the tetrapeptide. However, the transfer of the activated acetyl group to the ACP domain was not observed, probably attributed to the improper capability of Sfp to activate apo-ACP to the holo-ACP form. PMID- 18388459 TI - Enhanced enzyme activities of inclusion bodies of recombinant beta-galactosidase via the addition of inducer analog after L-arabinose induction in the araBAD promoter system of Escherichia coli. AB - We observed that an inclusion body (IB) of recombinant beta-galactosidase that was produced by the araBAD promoter system in Escherichia coli (E. coli) showed enzyme activity. In order to improve its activity, the lowering of the transcription rate of the beta-galactosidase structural gene was attempted through competition between an inducer (L-arabinose) and an inducer analog (D fucose). In the deep-well microtiter plate culture and lab-scale fermentor culture, it was demonstrated that the addition of D-fucose caused an improvement in specific beta-galactosidase production, although beta-galactosidase was produced as an IB. In particular, the addition of D-fucose after induction led to an increase in the specific activity of beta-galactosidase IB. Finally, we confirmed that the addition of D-fucose after induction caused changes in the structure of beta-galactosidase IB, with higher enzyme activity. Based on these results, we expect that an improved enzyme IB will be used as a biocatalyst of the enzyme bioprocess, because an enzyme IB can be purified easily and has physical durability. PMID- 18388460 TI - Thermostable beta-glycosidase-CBD fusion protein for biochemical analysis of cotton scouring efficiency. AB - Multidomain proteins for the biochemical analysis of the scouring efficiency of cotton fabrics were constructed by the fusion of a reporter moiety in the N terminal and the cellulose binding domain (CBD) in the C-terminal. Based on the specific binding of the CBD of Cellulomonas fimi exoglucanase (Cex) to crystalline cellulose (Avicel), the reporter protein is guided to the cellulose fibers that are increasingly exposed as the scouring process proceeds. Among the tested reporter proteins, a thermostable beta-glycosidase (BglA) from Thermus caldophilus was found to be most appropriate, showing a higher applicability and stability than GFP, DsRed, or a tetrameric beta-glucuronidase (GUS) from Escherichia coli, which were precipitated more seriously during the expression and purification steps. When cotton fabrics with different scouring levels were treated with the BglA-CBD and incubated with X-Gal as the chromogenic substrate, an indigo color became visible within 2 h, and the color depth changed according to the conditions and extent of the scouring. PMID- 18388461 TI - Molecular cloning and characterization of a bile salt hydrolase from Lactobacillus acidophilus PF01. AB - Phenotypic screening for bile salt hydrolase (BSH) activity was performed on Lactobacillus acidophilus PF01 isolated from piglet feces. A gene encoding BSH was identified and cloned from the genomic library of L. acidophilus PF01. The bsh gene and surrounding regions were characterized by nucleotide sequence analysis and were found to contain a single open reading frame (ORF) of 951 nucleotides encoding a 316 amino acid protein. The potential bsh promoter region was located upstream of the start codon. The protein deduced from the complete ORF had high similarity with other BSHs, and four amino acid motifs located around the active site, FGRNXD, AGLNF, VLTNXP, and GXGXGXXGXPGD, were highly conserved. The bsh gene was cloned into the pET21b expression vector and expressed in Escherichia coli BLR(DE3) by induction with 0.1mM of isopropylthiogalactopyranoside. The BSH enzyme was purified with apparent homogeneity using a Ni2+-NTA agarose column and characterized. The overexpressed recombinant BSH enzyme of L. acidophilus PF01 exhibited hydrolase activity against tauroconjugated bile salts, but not glycoconjugated bile salts. It showed the highest activity against taurocholic acid. The maximum BSH activity occurred at approximately 40oC. The enzyme maintained approximately 70% of its maximum activity even at 60 degrees , whereas its activity rapidly decreased at below 37 degrees . The optimum pH was 6, and BSH activity was rapidly inactivated below pH 5 and above pH 7. PMID- 18388462 TI - Purification and characterization of branching specificity of a novel extracellular amylolytic enzyme from marine hyperthermophilic Rhodothermus marinus. AB - An extracellular enzyme (RMEBE) possessing alpha- (1-->4)-(1-->6)-transferring activity was purified to homogeneity from Rhodothermus marinus by combination of ammonium sulfate precipitation, Q-Sepharose ion-exchange, and Superdex- 200 gel filtration chromatographies, and preparative native polyacrylamide gel electrophoresis. The purified enzyme had an optimum pH of 6.0 and was highly thermostable with a maximal activity at 80 degrees . Its half-life was determined to be 73.7 and 16.7 min at 80 and 85 degrees , respectively. The enzyme was also halophilic and highly halotolerant up to about 2 M NaCl, with a maximal activity at 0.5M. The substrate specificity of RMEBE suggested that it possesses partial characteristics of both glucan branching enzyme and neopullulanase. RMEBE clearly produced branched glucans from amylose, with partial alpha-(1-->4)-hydrolysis of amylose and starch. At the same time, it hydrolyzed pullulan partly to panose, and exhibited alpha-(1-->4)-(1-->6)-transferase activity for small maltooligosaccharides, producing disproportionated alpha-(1-->6)-branched maltooligosaccharides. The enzyme preferred maltopentaose and maltohexaose to smaller maltooligosaccharides for production of longer branched products. Thus, the results suggest that RMEBE might be applied for production of branched oligosaccharides from small maltodextrins at high temperature or even at high salinity. PMID- 18388463 TI - Development of magnetically separable immobilized lipase by using cellulose derivatives and their application in enantioselective esterification of ibuprofen. AB - Highly active, stable, and magnetically separable immobilized enzymes were developed using carboxymethyl cellulose (CMC) and diethylaminoethyl cellulose DEAE-C; hereafter designated "DEAE" as supporting materials. Iron oxide nanoparticles penetrated the micropores of the supporting materials, rendering them magnetically separable. Lipase (LP) was immobilized on the surface of the supporting materials by using cross-linked enzyme aggregation (CLEA) by glutaraldehyde. The activity of enzyme aggregates coated on DEAE was approximately 2 times higher than that of enzyme aggregates coated on CMC. This is explained by the fact that enzyme aggregates with amine residues are more efficient than those with carboxyl residues. After a 96-h enantioselective ibuprofen esterification reaction, 6% ibuprofen propyl ester was produced from the racemic mixture of ibuprofen by using DEAE-LP, and 2.8% using CMC-LP. PMID- 18388464 TI - Analysis of the dimerization of human CD99 using bimolecular fluorescence complementation technique. AB - Two isoforms of human CD99 have been identified, but only heterotypic interaction between the isomers was recently demonstrated. In this study, we performed bimolecular fluorescence complementation analysis to further characterize the interaction in vivo. Upon transiently transfecting plasmids expressing either of the two isoforms fused with yellow fluorescent protein (YFP) fragments, all the YFPtagged CD99 molecules were properly localized on cell surfaces, and formed fluorescent dimers. Interestingly, however, unlike the previous report, the homodimers formed as efficiently as the heterodimer via their extracellular domains, implying its distinct regulatory role through modulating the complex profile. PMID- 18388465 TI - Comparative biodegradation of HDPE and LDPE using an indigenously developed microbial consortium. AB - A variety of bacterial strains were isolated from waste disposal sites of Uttaranchal, India, and some from artificially developed soil beds containing maleic anhydride, glucose, and small pieces of polyethylene. Primary screening of isolates was done based on their ability to utilize high- and low-density polyethylenes (HDPE/LDPE) as a primary carbon source. Thereafter, a consortium was developed using potential strains. Furthermore, a biodegradation assay was carried out in 500-ml flasks containing minimal broth (250 ml) and HDPE/ LDPE at 5 mg/ml concentration. After incubation for two weeks, degraded samples were recovered through filtration and subsequent evaporation. Fourier transform infrared spectroscopy (FTIR) and simultaneous thermogravimetric-differential thermogravimetry-differential thermal analysis TG-DTG-DTA) were used to analyze these samples. Results showed that consortium-treated HDPE (considered to be more inert relative to LDPE) was degraded to a greater extent 22.41% weight loss) in comparison with LDPE (21.70% weight loss), whereas, in the case of untreated samples, weight loss was more for LDPE than HDPE (4.5% and 2.5%, respectively) at 400 degrees . Therefore, this study suggests that polyethylene could be degraded by utilizing microbial consortia in an eco-friendly manner. PMID- 18388466 TI - Selection of newly isolated mushroom strains for tolerance and biosorption of zinc in vitro. AB - Nine newly isolated mushroom strains were tested to assess both their zinc tolerance and potential for zinc removal from an aqueous solution. Four strains of ectomycorrhizal fungi, namely Clavariadelphus truncatus (T 192), Rhizopogon roseolus (T 21), Lepista nuda (T 373), and Tricholoma equestre (T 174), along with five strains of white rot fungi, Lenzites betulina (S 2), Trametes hirsuta (T 587), Ganoderma spp. (T 99), Polyporus arcularius (T 438), and Ganoderma carnosum (M 88), were investigated using zinc-amended solid and liquid media. Their biosorption properties were also determined. The colony diameter and dry weight were used as tolerance indices for fungal growth. C. truncatus and T. equestre were not strongly inhibited at the highest concentrations of (225 mg/l) zinc in solid media. The most tolerant four strains with solid media, C. truncatus, G. carnosum, T. hirsuta, and T. equestre, were then chosen for tolerance tests in liquid media. An ectomycorrhizal strain, C. truncatus, was also detected as the most tolerant strain in liquid media. However, the metal tolerant strains demonstrated weak activity in the biosorption studies. In contrast, the highest biosorption activity was presented by a more sensitive strain, G. carnosum. In addition, seven different biosorbent types from G. carnosum (M 88) were compared for their Zn (II) biosorption in batch experiments. PMID- 18388467 TI - Characterization of growth-supporting factors produced by Geobacillus toebii for the commensal thermophile Symbiobacterium toebii. AB - Symbiobacterium toebii is a commensal symbiotic thermophile that cannot grow without support from a partner bacterium. We investigated the properties of Symbiobacterium growth-supporting factors (SGSFs) produced by the partner bacterium Geobacillus toebii. SGSFs occurred in both the cell-free extract (CFE) and culture supernatant of G. toebii and might comprise multifarious materials because of their different biological properties. The heavy SGSF contained in the cytosolic component exhibited heat- and proteinase-sensitive proteinaceous properties and had a molecular mass of >50 kDa. In contrast, the light SGSF contained in the extracellular component exhibited heat-stable, proteinase resistant, nonprotein properties and had a molecular mass of <10 kDa. Under morphological examination using light microscopy, S. toebii cultured with the culture supernatant of G. toebii was filamentous, whereas S. toebii cultured with the CFE of G. toebii was rod-shaped. These results strongly suggest that the SGSFs produced by G. toebii comprise two or more types that differ in their growth-supporting mechanisms, although all support the growth of S. toebii. Upon the examination of the distribution of SGSFs in other bacteria, both cytosolic and extracellular components of Geobacillus kaustophilus, Escherichia coli, and Bacillus subtilis had detectable growth-supporting effects for S. toebii, indicating that common SGSF materials are widely present in various bacterial strains. PMID- 18388468 TI - Chemical composition and antimicrobial activity of essential oil from cones of Pinus koraiensis. AB - The essential oil from the cones of Pinus koraiensis was prepared after removing the seeds, and its chemical composition analyzed using gas chromatography-mass spectrometry (GC-MS). Hydrodistillation of the P. koraiensis cones yielded 1.07% (v/w) of essential oil, which was almost three times the amount of essential oil extracted from the needles of the same plant. Moreover, the antimicrobial activities of the oil against the growth of Gram-positive bacteria, Gram-negative bacteria, and fungi were evaluated using the agar disc diffusion method and broth microdilution method. Eighty-seven components, comprising about 96.8% of the total oil, were identified. The most abundant oil components were limonene (27.90%), alpha-pinene (23.89%), beta-pinene (12.02%), 3-carene (4.95%), beta myrcene (4.53%), isolongifolene (3.35%), (-)-bornyl acetate (2.02%), caryophyllene (1.71%), and camphene (1.54%). The essential oil was confirmed to have significant antimicrobial activities, especially against pathogenic fungal strains such as Candida glabrata YFCC 062 and Cryptococcus neoformans B 42419. Therefore, the present results indicate that the essential oil from the cones of Pinus koraiensis can be used in various ways as a nontoxic and environmentally friendly disinfectant. PMID- 18388469 TI - Evaluation of biomolecular interactions of sulfated polysaccharide isolated from Grateloupia filicina on blood coagulation factors. AB - An edible marine red alga, Grateloupia filicina, collected from Jeju Island of Korea was hydrolyzed by cheap food-grade carbohydrases (Viscozyme, Celluclast, AMG, Termamyl, and Ultraflo) to investigate its anticoagulant activity. Among the tested enzymatic extracts of G. filicina, a Termamyl extract showed the highest anticoagulant activity. Anion-exchange chromatography on DEAE-cellulose and gelpermeation chromatography on Sepharose-4B were used to purify the active polysaccharide from the crude polysaccharide fraction of G. filicina. The purified sulfated polysaccharide (0.42 sulfate/total sugar) showed approximately 1,357 kDa molecular mass and was comprised mainly of galactose (98%) and 1-2% of glucose. The sample showed potential anticoagulant activity on activated partial thromboplastin time (APTT) and thrombin time (TT) assays. The purified G. filicina anticoagulant (GFA) inhibited the coagulation factor X (92%), factor II (82%), and factor VII (68%) of the coagulation cascade, and the molecular interaction (protein-polysaccharide) was highly enhanced in the presence of ATIII (antithrombin III). The dissociation constant of polysaccharide towards serine proteins decreased in the order of FXa (58.9 nM) >FIIa (74.6 nM) >FVII (109.3 nM). The low/less cytotoxicity of the polysaccharide benefits its use in the pharmaceutical industry; however, further studies that would help us to elucidate the mechanism of its activity are needed. PMID- 18388470 TI - Apoptosis of human hepatocarcinoma (HepG2) and neuroblastoma (SKN-SH) cells induced by polysaccharides-peptide complexes produced by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala. AB - Three different polysaccharide-peptide complexes (PPC, named as Fr-I, Fr-II, and Fr-III) were produced by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala, and their anticancer activities were investigated in human hepatocarcinoma (HepG2) and neuroblastoma (SK-N-SH) cells. The highest inhibitory effects of PPC on both HepG2 and SK-N-SH cells were achieved with Fr I, whereas Fr-III with low molecular mass showed lower inhibition effects. Interestingly, the inhibitory effects of the three fractions were increased after protease digestion, suggesting that the inhibitory effects resulted mainly from the carbohydrate moiety, at least in the case of Fr-II and Fr-III, of PPC. The results of DNA fragmentation in PPC-induced apoptotic cells were confirmed by both DNA ladder assay and comet assay. Our investigation also showed that PPC induced apoptosis of both cancer cells was associated with intracellular events including DNA fragmentation, activation of caspase-3, and modulation of Bcl-2 and Bax. We conclude that PPC has potential as a novel therapeutic agent for the treatment of both HepG2 and SK-N-SH cancer cells without any cytotoxicity against normal cells. PMID- 18388471 TI - Soraphinol C, a new free-radical scavenger from Sorangium cellulosum. AB - A new compound named soraphinol C (1) was isolated from myxobacteria Sorangium cellulosum KM1001 a soil isolate, together with a structurally related known compound, 4-hydroxysattabacin (2). These compounds were isolated by silica gel column chromatography and recycling preparative HPLC, consecutively. The structures of the compounds were determined on the basis of combined spectroscopic analyses. Compounds 1 and 2 exhibited antioxidant activity as a radical scavenger in the experiment using a hydrophilic free-radical initiator, 2,2'-azobis(2-amidinopropane)dihydrochloride with ORAC values of 0.956 and 0.617, respectively. PMID- 18388472 TI - Sensitization of the apoptotic effect of gamma-irradiation in genistein pretreated CaSki cervical cancer cells. AB - Radiotherapy is currently applied in the treatment of human cancers. We studied whether genistein would enhance the radiosensitivity and explored its precise molecular mechanism in cervical cancer cells. After co-treatment with genistein and irradiation, the viability, cell cycle analysis, and apoptosis signaling cascades were elucidated in CaSki cells. The viability was decreased by co treatment with genistein and irradiation compared with irradiation treatment alone. Treatment with only gamma-irradiation led to cell cycle arrest at the G1 phase. On the other hand, co-treatment with genistein and gamma-irradiation caused a decrease in the G1 phase and a concomitant increase up to 56% in the number of G2 phase. In addition, cotreatment increased the expression of p53 and p21, and Cdc2- tyr-15-p, supporting the occurrence of G2/M arrest. In general, apoptosis signaling cascades were activated by the following events: release of cytochrome c, upregulation of Bax, downregulation of Bcl-2, and activation of caspase-3 and -8 in the treatment of genistein and irradiation. Apparently, co treatment downregulated the transcripts of E6*I, E6*II, and E7. Genistein also stimulated irradiation-induced intracellular reactive oxygene, species (ROS) production, and co-treatment-induced apoptosis was inhibited by the antioxidant N acetylcysteine, suggesting that apoptosis has occurred through the increase in ROS by genistein and gamma-irradiation in cervical cancer cells. Gamma irradiation increased cyclooxygenase-1 (COX-2) expression, whereas the combination with genistein and gamma-irradiation almost completely prevented irradiation-induced COX-2 expression and PGE2 production. Co-treatment with genistein and gamma-irradiation inhibited proliferation through G2/M arrest and induced apoptosis via ROS modulation in the CaSki cancer cells. PMID- 18388473 TI - Carbon and energy balances of glucose fermentation with hydrogenproducing bacterium Citrobacter amalonaticus Y19. AB - For the newly isolated H2-producing chemoheterotrophic bacterium Citrobacter amalonaticus Y19, anaerobic glucose metabolism was studied in batch cultivation at varying initial glucose concentrations (3.5- 9.5 g/l). The carbon-mass and energy balances were determined and utilized to analyze the carbon metabolic pathways network. The analyses revealed (a) variable production of major metabolites (H2, ethanol, acetate, lactate, CO2, and cell mass) depending on initial glucose levels; (b) influence of NADH regeneration on the production of acetate, lactate, and ethanol; and (c) influence of the molar production of ATP on the production of biomass. The results reported in this paper suggest how the carbon metabolic pathway(s) should be designed for optimal H2 production, especially at high glucose concentrations, such as by blocking the carbon flux via lactate dehydrogenase from the pyruvate node. PMID- 18388474 TI - Enhanced biomass and gamma-linolenic acid production of mutant strain Arthrospira platensis. AB - A mutant of Arthrospira platensis PCC 9108, strain M9108, obtained by mutagenesis with UV treatment, was able to mixotrophically grow in an SOT medium containing 40 g of glucose/l. The biomass and specific growth rate of strain M9108 (4.10 g/l and 0.70/d) were 1.9-fold and 1.4-fold higher, respectively, than those of the wild type (2.21 g/l and 0.58/d) under mixotrophic culture condition. In addition, when compared with the wild type, the content of gamma- linolenic acid (GLA) in the mutant was increased when glucose concentration was increased. Compared with the wild type, the GLA content of the mutant was 2-fold higher in autotrophic culture and about 3-fold higher in mixotrophic culture. Thus, the mutant appears to possess more efficient facility to assimilate and metabolize glucose and to produce more GLA than its wild-type strain. PMID- 18388475 TI - Production of ethanol directly from potato starch by mixed culture of Saccharomyces cerevisiae and Aspergillus niger using electrochemical bioreactor. AB - When cultivated aerobically, Aspergillus niger hyphae produced extracellular glucoamylase, which catalyzes the saccharification of unliquified potato starch into glucose, but not when grown under anaerobic conditions. The Km and Vmax of the extracellular glucoamylase were 652.3 mg starch l-1 and 253.3 mg glucose l-1 min-1, respectively. In mixed culture of A. niger and Saccharomyces cerevisiae, oxygen had a negative influence on the alcohol fermentation of yeast, but activated fungal growth. Therefore, oxygen is a critical factor for ethanol production in the mixed culture, and its generation through electrolysis of water in an electrochemical bioreactor needs to be optimized for ethanol production from starch by coculture of fungal hyphae and yeast cells. By applying pulsed electric fields (PEF) into the electrochemical bioreactor, ethanol production from starch improved significantly: Ethanol produced from 50 g potato starch l-1 by a mixed culture of A. niger and S. cerevisiae was about 5 g l-1 in a conventional bioreactor, but was 9 g l-1 in 5 volts of PEF and about 19 g l-1 in 4 volts of PEF for 5 days. PMID- 18388476 TI - R-stereoselective amidase from Rhodococcus erythropolis No. 7 acting on 4-chloro 3-hydroxybutyramide. AB - Ethyl (S)-4-chloro-3-hydroxybutyrate is an intermediate for the synthesis of Atorvastatin, a chiral drug used for hypercholesterolemia. A Rhodococcus erythropolis strain (No. 7) able to convert 4-chloro-3-hydroxybutyronitrile into 4-chloro-3-hydroxybutyric acid has recently been isolated from soil. This activity has been regarded as having been caused by the successive actions of the nitrile hydratase and amidase. In this instance, the corresponding amidase gene was cloned from the R. erythropolis strain and expressed in Escherichia coli cells. A soluble active form of amidase enzyme was obtained at 18 degrees . The Ni column-purified recombinant amidase was found to have a specific activity of 3.89 U/mg toward the substrate isobutyramide. The amidase was found to exhibit a higher degree of activity when used with midchain substrates than with short chain ones. Put differently, amongst the various amides tested, isobutyramide and butyramide were found to be hydrolyzed the most rapidly. In addition to amidase activity, the enzyme was found to exhibit acyltransferase activity when hydroxyl amine was present. This dual activity has also been observed in other enzymes belonging to the same amidase group (E.C. 3.5.1.4). Moreover, the purified enzyme was proven to be able to enantioselectively hydrolyze 4-chloro-3 hydroxybutyramide into the corresponding acid. The e.e. value was measured to be 52% when the conversion yield was 57%. Although this e.e. value is low for direct commercial use, molecular evolution could eventually result in this amidase being used as a biocatalyst for the production of ethyl (S)-4-chloro-3-hydroxybutyrate. PMID- 18388477 TI - Factors influencing the production of water-soluble endopolysaccharides and exopolysaccharides from Lentinus lepideus and their effects on immune cytokine production. AB - An efficient method to produce water-soluble polysaccharides from Lentinus lepideus is described. The productivity of both endopolysaccharides (PPS) and exopolysaccharides (EPS) was compared under various culture conditions. The effect of treating their own PPS and EPS on immune cytokine production was also studied in relation to culture factors. High yield production of EPS required moderate culture temperature (25 degrees ) as well as long culture period (16-20 days). In contrast, PPS production required high culture temperature (30 degrees ) and short culture period ( days). Most of the carbon sources did not affect polysaccharides and mycelial production except for sucrose. Immune cytokine levels in the EPS treatment varied among carbon sources or culture periods. PPS did not appear to affect much on the production of cytokines, regardless of the culturing factors, except for the culture period. These results suggest that the optimal culture conditions for L. lepideus vary according to culture purposes, and different culture conditions should be used for different targets including mycelial biomass, EPS, and PPS. Whereas the immunomodulating activity of EPS appeared to be affected by culture conditions in L. lepideus, that of PPS did not. PMID- 18388478 TI - Antimicrobial (BN/PE) film combined with modified atmosphere packaging extends the shelf life of minimally processed fresh-cut iceberg lettuce. AB - This study was conducted to investigate the effect of modified atmosphere packaging (MAP) in combination with BN/PE film on the shelf life and quality of fresh-cut iceberg lettuce during cold storage. The total mesophilic population in the sample packed in BN/PE film under MAP conditions was dramatically reduced in comparison with that of PE film, PE film under MAP conditions, and BN/PE film. The O2 concentration in the BN/PE film under MAP conditions decreased slightly as the storage period progressed. The coloration of the iceberg lettuce progressed the slowest when it was packaged in BN/PE film under MAP conditions, followed by BN/PE film, PE film, and PE film under MAP conditions. The shelf life of fresh cut iceberg lettuce packaged in the BN/PE film under MAP conditions was extended by more than 2 days at 10 degrees as compared with that of the BN/PE film in which the extension effect was more than 2 days longer than that of PE, PET, and OPP films. PMID- 18388479 TI - Hydrolysis of isoflavone glucosides in soymilk fermented with single or mixed cultures of Lactobacillus paraplantarum KM, Weissella sp. 33, and Enterococcus faecium 35 isolated from humans. AB - Lactobacillus paraplantarum KM (Lp), Weissella sp. 33 (Ws), and Enterococcus faecium 35 (Ef) were used in single (Lp, Ws, Ef) or mixed cultures (Lp+Ws, Lp+Ef, Ws+Ef) for soymilk fermentation (37 degrees C, 12 h). After 12 h, the cell numbers, pH, and TA of soymilk were 7.4x108 -6.0x109 CFU/ ml, 3.8-4.5, and 0.59 0.70%, respectively. Changes in the contents of glycitin and genistin in soymilk fermented with Ef were not significant (p<0.05). The contents of isoflavone glucosides in soymilk fermented with the other cultures decreased significantly with an increase of aglycone contents (p<0.05). It corresponded well with a sharp increase in beta- glucosidase activity during fermentation. About 92-100% of the daidzin and 98-100% of the genistin in soymilk were converted to corresponding aglycones by Lp, Ws, or Lp+Ef within 12 h. PMID- 18388480 TI - Evaluation of a chromogenic medium supplemented with glucose for detecting Enterobacter sakazakii. AB - A commercial chromogenic agar medium (DFI) was supplemented with glucose (mDFI) to enhance the specificity of Enterobacter sakazakii (E. sakazakii) detection. Escherichia vulneris (E. vulneris), a putative false-positive strain on the DFI medium, produces alpha-glucosidase. The enzyme alpha- glucosidase hydrolyzes a substrate, 5-bromo-4-chloro-3- indolyl-alpha,D-glucopyranoside (XalphaGlc), producing green colonies. E. sakazakii strains produced green colonies on both DFI and mDFI agar, whereas E. vulneris produced green colonies on DFI agar but small white colonies on mDFI agar. E. sakazakii and E. vulneris were also readily differentiated by colony color when the mixed culture of the two strains was plated on mDFI agar and incubated for 24 h at 37 degrees C. The results indicate that the selectivity of the commercial chromogenic agar medium could be improved by a simple supplementation with glucose. PMID- 18388481 TI - Analysis of natural recombination in porcine endogenous retrovirus envelope genes. AB - Human tropic Porcine Endogenous Retroviruses (PERVs) are the major concern in zoonosis for xenotransplantation because PERVs cannot be eliminated by specific pathogen-free breeding. Recently, a PERV A/C recombinant with PERV-C bearing PERV A gp70 showed a higher infectivity (approximately 500-fold) to human cells than PERV-A. Additionally, the chance of recombination between PERVs and HERVs is frequently stated as another risk of xenografting. Overcoming zoonotic barriers in xenotransplantation is more complicated by recombination. To achieve successful xenotransplantation, studies on the recombination in PERVs are important. Here, we cloned and sequenced proviral PERV env sequences from pig gDNAs to analyze natural recombination. The envelope is the most important element in retroviruses as a pivotal determinant of host tropisms. As a result, a total of 164 PERV envelope genes were cloned from pigs (four conventional pigs and two miniature pigs). Distribution analysis and recombination analysis of PERVs were performed. Among them, five A/B recombinant clones were identified. Based on our analysis, we determined the minimum natural recombination frequency among PERVs to be 3%. Although a functional recombinant envelope clone was not found, our data evidently show that the recombination event among PERVs may occur naturally in pigs with a rather high possibility. PMID- 18388482 TI - Protective immunity induced by systemic and mucosal delivery of DNA vaccine expressing glycoprotein B of pseudorabies virus. AB - A murine model immunized by systemic and mucosal delivery of plasmid DNA vaccine expressing glycoprotein B (pCIgB) of pseudorabies virus (PrV) was used to evaluate both the nature of the induced immunity and protection against a virulent virus. With regard to systemic delivery, the intramuscular (i.m.) immunization with pCIgB induced strong PrV-specific IgG responses in serum but was inefficient in generating a mucosal IgA response. Mucosal delivery through intranasal (i.n.) immunization of pCIgB induced both systemic and mucosal immunity at the distal mucosal site. However, the levels of systemic immunity induced by i.n. immunization were less than those induced by i.m. immunization. Moreover, i.n. genetic transfer of pCIgB appeared to induce Th2-biased immunity compared with systemic delivery, as judged by the ratio of PrV-specific IgG isotypes and Th1- and Th2-type cytokines produced by stimulated T cells. Moreover, the immunity induced by i.n. immunization did not provide effective protection against i.n. challenge of a virulent PrV strain, whereas i.m. immunization produced resistance to viral infection. Therefore, although i.n. immunization was a useful route for inducing mucosal immunity at the virus entry site, i.n. immunization did not provide effective protection against the lethal infection of PrV. PMID- 18388483 TI - Role of CAGE, a novel cancer/testis antigen, in various cellular processes, including tumorigenesis, cytolytic T lymphocyte induction, and cell motility. AB - A cancer-associated antigen gene (CAGE) was identified by serological analysis of a recombinant cDNA expression library (SEREX). The gene was identified by screening cDNA expression libraries of human testis and gastric cancer cell lines with sera from patients with gastric cancer. CAGE was found to contain a D-E-A-D box domain and encodes a putative protein of 630 amino acids with possible helicase activity. The CAGE gene is widely expressed in various cancer tissues and cancer cell lines. Demethylation plays a role in the activation of CAGE in certain cancer cell lines where the gene is not expressed. The functional roles of CAGE in tumorigenesis, the molecular mechanisms of CAGE expression, and cell motility are also discussed. PMID- 18388484 TI - Ribosomal catalysis: the evolution of mechanistic concepts for peptide bond formation and peptidyl-tRNA hydrolysis. AB - Over time the mechanistic concepts to describe the two principal chemical reactions that are catalyzed by the ribosome, peptide bond formation and peptidyl tRNA hydrolysis, have undergone dramatic changes. While the initial models were based on a ribosomal protein-based mechanism, evidence for a direct functional contribution of ribosomal RNA for catalysis has accumulated over the past years. The presentation of high resolution crystallographic structures of the large ribosomal subunit at the beginning of the new millennium dramatically increased our molecular insight into the organization of the active center and finally placed the ribosome amongst the list of RNA enzymes. Combined with elaborate biochemical and biophysical approaches the translation field has made significant progress in understanding mechanistic details of ribosomal catalysis. While it seems that the mechanism of ribosome-catalyzed peptidyl-tRNA hydrolysis is just emerging, the knowledge on transpeptidation is already very advanced. It has been realized that intricate interactions between ribosomal RNA and the transfer RNA substrate are crucial for proton shuttling that is required for efficient amide bond formation. PMID- 18388485 TI - Riboswitch folding: one at a time and step by step. AB - Riboswitches are mRNAs that regulate gene expression upon specific binding of a small metabolite (vitamin cofactors, nucleobases, amino acids, carbohydrates or even metal ions). Riboswitches must fold into very intricate 3D structures to accomplish their function. Single-molecule studies are very powerful techniques that allow the characterization and elucidation of complex folding paths as well as the identification of intermediate conformational states that an RNA must traverse to acquire its native, functional structure. We review some recent reports on the folding of the adenine-binding riboswitch as studied by fluorescence resonance energy transfer (FRET) and force-measuring optical tweezers (FMOT) techniques. PMID- 18388486 TI - NMR-spectroscopic characterization of phosphodiester bond cleavage catalyzed by the minimal hammerhead ribozyme. AB - In order to relate the conformational dynamics of the hammerhead ribozyme to its biological function the cleavage reaction catalyzed by the hammerhead ribozyme was monitored by time-resolved nuclear magnetic resonance (NMR) spectroscopy. For this purpose, the two nucleosides around the scissile phosphodiester bond were selectively (13)C labelled in multi-step organic syntheses starting from uniformly (13)C-labelled glucose. The phosphoamidites were incorporated using phosphoamidite chemistry in the hammerhead substrate strand. In addition, the 2' OH group on the 5'-side of the hammerhead substrate strand was labelled with a photolabile protecting group. This labelling strategy enabled a detailed characterisation of the nucleotides around the scissile phosphodiester bond in the ground state conformation of the hammerhead ribozyme in the absence and presence of Mg(2+) ions as well as of the product state. Photochemical induction of the reaction in situ was further characterized by time-resolved NMR spectroscopy. The detailed structural and dynamic investigations revealed that the conformation of the hammerhead ribozyme is significantly affected by addition of Mg(2+) leading to an ensemble of conformations where dynamic transitions between energetically similar conformations occur on the ms-timescale in the presence of Mg(2+). The dynamic transitions are localized around the catalytic core. Cleavage from this ensemble cannot be described by mono-exponential kinetics but follows bi-exponential kinetics. A model is described to take into account these experimental data. PMID- 18388487 TI - Alternative splicing and disease. AB - Splicing is a molecular mechanism, by which introns are removed from an mRNA precursor and exons are ligated to form a mature mRNA. Mutations that cause defects in the splicing mechanism are known to be responsible for many diseases, including cystic fibrosis and familial dysautonomia. If mutations that cause defects in splicing are responsible for such severe deleterious phenotypic differences, it is possible that mutations in splicing are also responsible for mildly deleterious phenotypic differences. Although deleterious mutations are rapidly eliminated from the population by purifying selection, the selection against mild deleterious effects is not as strong. Since mildly deleterious mutations have a chance of surviving natural selection, we might be mistakenly referring to these mutations as neutral variation between individuals. Splicing has also been shown to be seriously affected in cancer. Examination of cancerous tissues revealed alterations in expression levels of genes involved in mRNA processing and also a slight reduction in the level of exon skipping--the most common form of alternative splicing in humans. This implies that defects in genes involved in the regulation of splicing in cancerous tissues affect the delicate regulation of the inclusion level of alternatively skipped exons, shifting their mode of splicing back to constitutive. It may be that splicing silencers play a more prominent role in alternative splicing regulation than previously anticipated. PMID- 18388488 TI - Collaborative vaccine development: partnering pays. AB - Vaccine development, supported by infusions of public and private venture capital, is re-entering a golden age as one of the fastest growing sectors in the life-sciences industry. Demand is driven by great unmet need in underdeveloped countries, increased resistance to current treatments, bioterrorism, and for prevention indications in travelers, pediatric, and adult diseases. Production systems are becoming less reliant on processes such as egg-based manufacturing, while new processes can help to optimize vaccines. Expeditious development hinges on efficient study conduct, which is greatly enhanced through research partnerships with specialized contract research organizations (CROs) that are licensed and knowledgeable in the intricacies of immunology and with the technologic and scientific foundation to support changing timelines and strategies inherent to vaccine development. The CRO often brings a more objective assessment for probability of success and may offer alternative development pathways. Vaccine developers are afforded more flexibility and are free to focus on innovation and internal core competencies. Functions readily outsourced to a competent partner include animal model development, safety and efficacy studies, immunotoxicity and immunogenicity, dose response studies, and stability and potency testing. These functions capitalize on the CRO partner's regulatory and scientific talent and expertise, and reduce infrastructure expenses for the vaccine developer. Successful partnerships result in development efficiencies, elimination or reduced redundancies, and improved time to market. Keys to success include honest communications, transparency, and flexibility. PMID- 18388489 TI - How do ncRNAs guide chromatin-modifying complexes to specific locations within the nucleus? AB - Transcriptome analyses have led to the realization that eukaryotic cells make a large number of noncoding RNAs (ncRNAs). It appears that some of these are involved in guiding chromatin-modifying complexes to specific locations within the nucleus. How such ncRNAs function is largely unknown but various models have been proposed. Here we briefly discuss the evidence supporting two such models; that ncRNAs function by annealing either with nascent transcripts or with homologous DNA sequences. We then review a third model that is based on our recent work on the role of the noncoding roX RNAs in the localization of the MSL complex to sites on the X chromosome in Drosophila. Our results suggest that the MSL1 and MSL2 proteins bind to chromatin but it is the incorporation of the roX RNAs into the complex that somehow alters the binding specificity of the MSL1/MSL2 proteins to recognize sites on the X chromosome. PMID- 18388490 TI - Evolution of type-specific immunoassays to evaluate the functional immune response to Gardasil: a vaccine for human papillomavirus types 16, 18, 6 and 11. AB - Epidemiological studies and clinical trials of vaccines depend on the accurate measurement of antibodies within the polyclonal response to infection or vaccination. The assay currently used to measure the antibody response to vaccination with GARDASIL [Quadrivalent Human Papillomavirus (Types 6, 11, 16, 18) Recombinant Vaccine]--a quadrivalent vaccine used against human papillomavirus (HPV) types 6, 11, 16, and 18--is a competitive Luminex assay (cLIA) that uses multiplex technology to detect type-specific neutralizing antibodies against all four HPV types in a single serum sample. Here we describe how the cLIA was developed, as well as how the monoclonal antibodies (mAbs), used as competitors in the assay, were characterized. An enzyme-linked immunosorbent assay (ELISA) was used to screen eight previously-identified mAbs for their ability to bind to HPV virus-like particles (VLPs) in a type-specific and conformation-dependent manner. Four of these mAbs, H6.M48, K11.B2, H16.V5, and H18.J4, met our specifications and were shown to have the potential to neutralize HPV infection in hemagglutination inhibition and pseudovirus neutralization assays. The competitive immunoassay format was able to distinguish type-specific antibodies in the sera of nonhuman primates vaccinated with HPV VLPs, whereas a traditional direct-bind ELISA could not. In addition, the serum antibodies measured by the competitive assay are known to be neutralizing, whereas the ELISA does not distinguish neutralizing and nonneutralizing antibodies in a serum sample. By detecting antibodies to neutralizing epitopes, the competitive assay both demonstrates sero-conversion and provides a potential functional link between sero-conversion and protective immunity in response to vaccination with GARDASIL. PMID- 18388491 TI - Two distinct domains of Bruno bind specifically to the oskar mRNA. AB - Selective deployment of Oskar protein at the posterior pole of the Drosophila oocyte relies on localization of oskar mRNA, combined with translational regulation to ensure that only the localized mRNA produces protein. The Bruno protein binds to Bruno Response Elements (BREs) in the oskar mRNA, and prevents translation of unlocalized oskar mRNA. Bruno contains three copies of the RNA Recognition Motif (RRM), a protein motif that often binds directly to RNA. Either of two nonoverlapping parts of Bruno--RRMs 1 and 2, and RRM 3 and 42 flanking amino acids--can bind specifically to BRE-containing RNA, but both domains are required for maximal binding. When expressed in Drosophila ovaries, Bruno proteins with a single RNA binding domain mutated have reduced repressive activity, while mutation of both binding domains largely eliminates this activity. Notably, the same proteins expressed as fusions to GFP accumulate in nuclei, with the most severe mislocalization occurring when both RNA binding domains are mutated. A similar mislocalization of endogenous Bruno occurs when mRNA export is blocked. Thus, Bruno shuttles between the nucleus and cytoplasm, and may first bind oskar mRNA in the nucleus. PMID- 18388492 TI - Engineered riboswitches: overview, problems and trends. AB - The first conditional gene expression system which employed a small molecule binding aptamer was developed several years before the discovery of natural riboswitches. With the discovery of riboswitches it became obvious that nature uses exactly the same principal of direct RNA-ligand interaction to regulate gene expression in a highly efficient, precise and fast way. In the last decade, further engineered riboswitches have been developed to control gene expression in different organisms. The successful development of new engineered riboswitches, however, is not only dependent on an innovative design but also necessitates a two step process: first, an in vitro selection which results in aptamers with high affinity binding to a desired ligand and second, a subsequent screen to identify RNAs with a desired functionality within cells. This review will give an overview of recent reports of engineered riboswitches, highlight recent developments and point out trends and problems in the field. PMID- 18388493 TI - Detecting phosphorylation-dependent interactions with the C-terminal domain of RNA polymerase II subunit Rpb1p using a yeast two-hybrid assay. AB - Rpb1p, the largest subunit of S. cerevisiae RNA polymerase II, contains a repetitive structure called the C-terminal domain (CTD). The CTD serves as a scaffold for the regulated association and dissociation of more than a hundred proteins involved in RNA synthesis. Phosphorylation of two serine residues (Ser(2) and Ser(5)) in the repeating units of the CTD change dynamically during the pre-initiation, initiation, elongation and termination of transcription to control the binding and release of transcriptional components. A modification of the well established yeast two-hybrid assay for protein-protein interactions is described that detects interactions between phosphorylated forms of the CTD and proteins whose interactions with the CTD depend on phosphorylation. The efficacy of the approach was established by first showing that two-hybrid fusions containing the CTD are phosphorylated at Ser(2) and Ser(5) residues. Interactions between the CTD and three known CTD-binding proteins were analyzed. The results suggest that the modified two-hybrid system accurately assays CTD-binding and provides a new and convenient assay for CTD-binding proteins. PMID- 18388494 TI - Meningococcal B vaccine development and evaluation of efficacy. AB - In this commentary we stress that serum bactericidal activity is now internationally accepted as correlate of protection for evaluating new vaccines for epidemic and endemic meningococcal B control, and that the standardization of the assay and the choice of representative strains for each of the vaccine candidates are critical. PMID- 18388495 TI - Isolation of small RNA-binding proteins from E. coli: evidence for frequent interaction of RNAs with RNA polymerase. AB - Bacterial small RNAs (sRNAs) are non-coding RNAs that regulate gene expression enabling cells to adapt to various growth conditions. Assuming that most RNAs require proteins to exert their activities, we purified and identified sRNA binding factors via affinity chromatography and mass spectrometry. We consistently obtained RNA polymerase betasubunit, host factor Hfq and ribosomal protein S1 as sRNA-binding proteins in addition to several other factors. Most importantly, we observed that RNA polymerase not only binds several sRNAs but also reacts with them, both cleaving and extending the RNAs at their 3' ends. The fact that the RNA polymerase reacts with sRNAs maps their interaction site to the active centre cleft of the enzyme and shows that it takes RNAs as template to perform RNA-dependent RNA polymerase activity. We further performed genomic SELEX to isolate RNA polymerase-binding RNAs and obtained a large number of E. coli sequences binding with high affinity to this enzyme. In vivo binding of some of the RNAs to the RNA polymerase was confirmed via co-immunoprecipitation in cell extracts prepared from different growth conditions. Our observations show that RNA polymerase is able to bind and react with many different RNAs and we suggest that RNAs are involved in transcriptional regulation more frequently than anticipated. PMID- 18388496 TI - Corn trypsin inhibitor in fluorogenic thrombin-generation measurements is only necessary at low tissue factor concentrations and influences the relationship between factor VIII coagulant activity and thrombogram parameters. AB - The fluorogenic calibrated automated thrombin-generation assay is influenced by contact pathway activation in platelet-rich and platelet-poor plasma. This influence lessens with increasing tissue factor (TF) concentrations and is inhibited by corn trypsin inhibitor (CTI). CTI is expensive and at what TF concentration its influence becomes irrelevant is unclear. Spiking of factor VIII (FVIII)-depleted plasma with FVIII, in samples without CTI, shows a plateau of thrombin generation at low normal FVIII levels. Given the association with thrombosis at high levels, a continuing increase in thrombin generation would be expected. We studied the effect of CTI on this relation by spiking experiments up to 4.8 IU/ml at 1 pmol/l TF and compared the influence of CTI at 1 and 5 pmol/l in platelet-poor plasma. CTI significantly influences thrombin generation in platelet-poor plasma at 1 pmol/l TF (difference of means for endogenous thrombin potential of 232.5 nmol/l per min, P<0.0001) and peak of 48 nmol/l (P<0.0001)) but not at 5 pmol/l. Spiking experiments without CTI confirm the hyperbolic relation between FVIII coagulant activity (FVIII:C) and endogenous thrombin potential with a plateau at 0.70-1.40 IU/ml. With CTI, a near-linear response up to 1.0 IU/ml was found with a plateau at 2.4-4.8 IU/ml. For peak thrombin, no plateau was reached with CTI. The present study confirms and extends previous data on CTI and the relationship between FVIII:C and thrombin generation. CTI is not necessary at 5 pmol/l TF, and thrombin generation remains dependent on FVIII:C up to 4.8 IU/ml at 1 pmol/l with CTI. Higher levels than previously thought may be needed to normalize thrombin generation. PMID- 18388497 TI - Persistent factor VIII-dependent factor X activation on endothelial cells is independent of von Willebrand factor. AB - Endothelial cells are able to support the activation of coagulation factor X by activated factor IX in the presence of its cofactor, factor VIII. We have previously reported that this reaction is persistent on endothelial cells, but transient on activated platelets and phospholipid vesicles when activated factor X (Xa) is used as activator of factor VIII. Aim of the present study was to explore the influence of von Willebrand factor and that of the factor VIII activator, either factor Xa or thrombin, on the decay of factor X activation on the endothelial cell surface. Kinetics of factor X activation on human umbilical vein endothelial cells was compared with that on phospholipid vesicles employing purified coagulation factors from plasma as well as recombinant factor VIII variants. Employing factor Xa as factor VIII activator, rate constants for decay of membrane-bound factor X activation were consistently low on endothelial cells (0.02 min) as compared with phospholipid vesicles (0.2 min). Activation of factor VIII by thrombin resulted in two-fold increased decay rates. In the presence of excess of von Willebrand factor over factor VIII, decay rates were not significantly changed. Factor VIII variants with and without a Tyr to Phe substitution, which abolishes high-affinity binding to von Willebrand factor, displayed the same factor X activation decay kinetics. Although previous studies have shown that von Willebrand factor modulates factor VIII activation and stabilisation, this apparently does not affect the progression of factor X activation at the endothelium. PMID- 18388498 TI - Screening of mutations of hemophilia A in 40 Italian patients: a novel G-to-A mutation in intron 10 of the F8 gene as a putative cause of mild hemophilia A in southern Italy. AB - Hemophilia A is an X-linked bleeding disorder caused by widespread mutations in the human coagulation factor 8 gene. We have searched for mutations in factor 8 gene DNAs from 40 unrelated Italian patients with hemophilia A. All patients came from the same region (Calabria) and were followed-up at the same hemophilia center. Of the 40 patients, 20 (50%) had severe hemophilia A, 19 (47.5%) had moderate hemophilia A, and one (2.5%) had mild hemophilia A. All patients were first screened for the common intron 22 and intron 1 inversions. Inversion negative samples were screened for point mutations by direct sequencing of all coding regions and intron-exon boundaries of the factor 8 gene. Mutations previously reported as causative of hemophilia A were identified in 14 of the 40 patients. These included five (12.5%) intron 22 inversions, one (2.5%) small deletion, one (2.5%) small insertion and seven (17.5%) point mutations. In all patients with moderate and mild hemophilia A, a nucleotide change in the c.1538 18G>A in intron 10, not reported in the HAMSTeRS factor 8 mutation database (http://europium.csc.mrc.ac.uk/), was found. The G-to-A change predicts the appearance of a new acceptor splice site. We have also demonstrated that all patients share a common haplotype, suggesting that the mutation probably occurred in a single ancestor. In conclusion, we suggest that the c.1538-18G>A transition can be the putative mutation, which probably occurred in a common ancestor and then spread in neighbours, in patients with moderate-mild hemophilia A investigated in the present study. PMID- 18388499 TI - Early intervention with antithrombin III therapy to prevent progression of hepatic venoocclusive disease. AB - Venoocclusive disease (VOD) is the most frequent cause of early nonrelapse mortality among patients receiving high-dose chemoradiotherapy and hematopoietic stem cell transplantation. Endothelial injury of sinusoids and hepatic veins following chemotherapy is considered the initial event in the development of VOD. Activation of the coagulation cascade and inflammatory processes following endothelial injury results in a hypercoagulable state and a localized consumption of the natural anticoagulants, antithrombin III, protein C and protein S. The resultant coagulopathy can lead to multiorgan dysfunction and death. The objective was to retrospectively study the largest series of patients that has received antithrombin III for the treatment of VOD following hematopoietic stem cell transplantation. A total of 48 patients were diagnosed with VOD post hematopoietic stem cell transplantation (median age, 39 years; range, 1-69 years); 38 of the 48 received a nonradiation-based conditioning regimen and 21 of 48 received a transplant from an unrelated donor. Treatment was primarily directed at early intervention rather than prophylactic therapy to correct the antithrombin III deficiency associated with VOD. We attempted to achieve antithrombin III levels greater than 120%. There was no significant treatment related morbidity. The overall 100-day mortality for the treatment cohort was 17%, with 10% for the mild/moderate group and 39% for the severe group, respectively. In conclusion, the encouraging results of this study suggest that this antithrombin III treatment should be further considered in patients with severe VOD. PMID- 18388500 TI - The use of rituximab as an adjuvant for immune tolerance therapy in a hemophilia B boy with inhibitor and anaphylaxis to factor IX concentrate. AB - We describe a 10-year-old severe hemophilia B boy with a stop codon mutation of exon 2 in the factor IX gene who developed high inhibitor of 70 Bethesda units (BU) from 12 months of age after exposure to prothrombin complex concentrate for 14 days. The inhibitor spontaneously disappeared within 3 months. The patient, however, exhibited anaphylactic reaction to the administration of prothrombin complex concentrate and factor IX concentrate at ages 15 and 23 months, respectively. Although recombinant activated factor VII was alternatively given, he suffered from progressive hemophilic arthropathy. At the age of 10 years, the boy underwent desensitization to factor IX concentrate and could tolerate factor IX concentrate of 40 U/kg administered on day 9 of desensitization. Unfortunately, the inhibitor of 16 BU was detected on day 6 and rapidly increased to 180 BU on day 9 of desensitization. Rituximab 375 mg/m2 per week was therefore immediately initiated on day 10 and a total of four doses were given. The inhibitor gradually decreased to 21.5 BU after the fourth dose of rituximab. The daily factor IX concentrate administration of 40 U/kg was continued for 1 month and decreased to three times per week for another month, and then to once to twice per week for the remaining 14 months of desensitization. The patient was able to attend regular school and the most recent inhibitor ranged from 4.4 to 10 BU. No proteinuria or alteration of renal function was found. In conclusion, rituximab is a helpful adjuvant to immune tolerance therapy in a hemophilia B boy with inhibitor and anaphylaxis to factor IX concentrate. PMID- 18388501 TI - The value of rotation thromboelastometry to monitor disturbed perioperative haemostasis and bleeding risk in patients with cardiopulmonary bypass. AB - Rotation thromboelastometry (ROTEM) performed on whole-blood samples provides information on the contribution of fibrinogen and platelets to clot formation. Such measurements are believed superior to classical plasma coagulation measurements as a means of monitoring disturbed haemostasis. On-pump cardiac surgery is associated with high bleeding risk. The study objective was to obtain information on the frequency of abnormal values of ROTEM variables and to assess their value in estimating bleeding risk in such patients. We studied 150 patients undergoing elective on-pump cardiac surgery. We found a significant surgery induced decrease in haemostatic potential, with more abnormal ROTEM values in intrinsically activated coagulation (up to 50%) than in extrinsically activated coagulation (up to 27%) or the maximum clot firmness in FIBTEM (10%), a test measuring the contribution of fibrinogen. All ROTEM variables tend to normalize within 14-18 h postoperatively. Best positive predictive values and specificity for a postoperative blood loss above 600 ml were found for the clot formation time in extrinsically activated coagulation (71%/94%) and the maximum clot firmness in FIBTEM (73%/95%); these values were superior to the activated partial thromboplastin time or prothrombin time (56%/72% and 43%/5%, respectively). There was no relation between preoperative or early postoperative ROTEM values and intraoperative bleeding. ROTEM recorded a benefit of administration of platelet concentrates or fresh-frozen plasma, particularly when given postoperatively, on haemostasis. In contrast, intraoperative administration of red blood cells impaired haemostasis. ROTEM can provide a more detailed diagnostic basis enabling a focused therapy to cardiac surgery patients with high bleeding risk. PMID- 18388502 TI - Whole blood coagulation measured by modified thrombelastography (ROTEM) is impaired in infants with congenital heart diseases. AB - Patients with congenital heart disease (CHD) often do have a variety of coagulation abnormalities that results in bleeding diathesis. Our study aimed to determine the impact of cyanosis and CHD on modified thrombelastography parameters, compared with children without CHD. Preoperative blood samples were taken for TEM analyses from a total of 51 infants scheduled for surgery. The following groups were examined: normal patients without CHD, acyanotic patients with acyanotic CHD, and cyanotic patients with CHD and with preoperative hemoglobin values higher than 15 g dl(-1). Mean values of all patient groups as well as all individual values of normal patients were within their normal ranges. Within these limits, however, clots were significantly inferior in cyanotic patients (worse mean values of eight out of 10 measured TEM parameters representing the intrinsic, extrinsic, and plasmatic pathways of coagulation) and in acyanotic patients (two out of 10 TEM parameters). Individually, pathological TEM parameters were found in seven (41%) cyanotic patients (P=0.003; vs. normal patients) and in three (17%) acyanotic patients (P=0.01). More than one abnormal TEM coagulation parameter was found in four patients, all of them cyanotic patients. Hyperfibrinolysis was detected in one patient, a cyanotic patient. The present investigation confirms previous findings that in patients with CHD the heart defect itself compromises coagulation monitored with TEM, but in addition, we demonstrate that cyanosis and/or polycythemia exert the essential negative impact on hemostasis. Preoperative hyperfibrinolysis detected with TEM seems to play no important role. PMID- 18388503 TI - Antithrombotic prophylaxis during pregnancy in women with deficiency of natural anticoagulants. AB - Prevalence of obstetric complications in women with deficiency of natural anticoagulants is difficult, because these defects are very rare in general population. Furthermore, the available data on prophylactic intervention in such individuals to decrease the obstetric risk are also very limited. To evaluate in a setting of women with rare hereditary thrombophilias, deficiency of antithrombin, protein C or protein S whether thromboprophylaxis during pregnancy could affect foeto-maternal outcome. Retrospective cohort study in two Italian thrombosis centres: 32 women with an established hereditary deficiency of natural anticoagulants antithrombin, protein C or protein S were enrolled because of a history of unexplained foetal losses with or without venous thromboembolism. Overall, information on the management of 103 pregnancies was obtained and the outcome in the absence or in the presence of enoxaparin treatment was recorded. Live births were recorded in 18 women (56.2%). Ten women had early foetal losses, 16 intrauterine foetal death and six both. Eight pregnancies were treated with enoxaparin. Seven out of eight treated pregnancies [87.5%, 95% confidence interval (CI) 52.9-97.7] and 27 out of 95 not-treated pregnancies (28.4%, 95% CI 20.3-38.1) resulted in the delivery of a live newborn (Fisher exact test: P=0.002), with a risk of foetal loss in untreated pregnancies 3.1 times (95% CI 1.7-3.5) higher than in treated ones. In this setting of patients with rare causes of thrombophilia, antithrombotic prophylaxis during pregnancy improves foeto-maternal outcome. PMID- 18388504 TI - ABO blood group distribution and major cardiovascular risk factors in patients with acute myocardial infarction. AB - We aimed to investigate whether there is an association between ABO blood groups, cardiovascular risk factors and myocardial infarction (MI) in a Turkish cohort. Four hundred and seventy-six patients with acute ST elevation MI (mean age 56.7+/ 11.7; 80% men) and 203 age and sex matched healthy subjects were enrolled in the study. ABO blood group distribution of patients was compared with control group. Furthermore, in each ABO blood group, frequency of major cardiac risk factors was determined to find any correlation between blood groups and cardiovascular risk factors. The distribution of ABO blood groups in patients versus control group was A in 43.1 versus 44.3%, B in 15.1 versus 15.3%, AB in 10.7 versus 12.3% and O in 31.1 versus 28.1% (P>0.05 for all). ABO blood group distribution of both patients and control group was concordant with the official data from general Turkish population. The frequency of cardiovascular risk factors was similar in patients with different blood groups; however, the patients with blood group A were younger (P=0.004) and coronary artery disease detection age was lower (P=0.001) than those with the other blood groups. The distribution of ABO blood groups in patients with MI was quite similar to that in control group and that of general Turkish population, which supports the idea that ABO blood group might not be significantly associated with the development of MI. Association of ABO blood group distribution with cardiovascular risk factors, coronary artery disease and MI needs to be clarified with multicenter, prospective and large scale studies. PMID- 18388505 TI - Major clinical vascular events and aspirin-resistance status as determined by the PFA-100 method among patients with stable coronary artery disease: a prospective study. AB - Aspirin inhibits platelet activation and reduces major vascular events in patients with stable coronary artery disease. The extent of platelet inhibition, denoted as aspirin resistance, however, is not always sufficient. A correlation between aspirin resistance as measured by aggregometry and adverse clinical events has been demonstrated. The point-of-care platelet function analyzer PFA 100 is usually used to detect aspirin resistance, but the relation between PFA 100 results and the vascular prognosis is not assessed. We prospectively enrolled 97 patients with stable coronary artery disease who were on aspirin (160 mg per day since 1 month or longer). Aspirin resistance was measured by the PFA-100 analyzer. Median follow-up was 2.5 years and the primary outcome was the composite of death, myocardial infarction, and ischemic cerebral infarction or acute limb ischemia. In our study, 29 patients (29.9%) showed resistance to aspirin, with a higher percentage of female patients (38 vs. 15%; P=0.01). During the follow-up, aspirin resistance was not associated with an increased risk of death, myocardial infarction, or ischemic vascular event compared with the aspirin-sensitive patients (17 vs. 13%; P>0.60). In this cohort of stable coronary artery disease, patients on aspirin dose of 160 mg per day, the aspirin resistance status based on the PFA-100 results is not associated with a significant increase in major vascular clinical events. PMID- 18388506 TI - Four novel FXI gene mutations in three factor XI- deficient patients. AB - Hereditary factor XI deficiency is a mild bleeding disorder, which is highly prevalent among Ashkenazi Jews, but has been reported in all populations. In Ashkenazi Jews, two factor XI gene mutations Glu 117X (type II) and Phe283Leu (type III) are particularly common. In other ethnic groups, factor XI deficiency is a rare bleeding disorder and is related to a variety of mutations throughout the factor XI gene. Three cases of quantitative factor XI deficiency in relation with four novel missense mutations are reported: a compound heterozygosity for two novel mutations (Ala 181 Val and Ala 412 Thr) with a severe factor XI deficiency and two missense mutations (His 388 Pro and Trp 407 Cys) in heterozygous patients with partial factor XI deficiency. PMID- 18388507 TI - Acute myocardial infarction, ischemic cerebrovascular disease and variceal bleeding due to portal vein thrombosis in a patient with hereditary thrombophilia. AB - We report on a 43-year-old female patient with multiple thrombotic risk factors who, in a few months, developed acute myocardial infarction, an ischemic cerebrovascular event and variceal bleeding due to portal vein thrombosis. The factor V Leiden mutation was carried in heterozygous form, homocysteine was elevated at 19.6 micromol/l, and methylenetetrahydrofolate reductase C677T mutation was carried in homozygous form. Moderately increased plasma homocysteine level and a reduced protein S activity were evident. Anticardiolipin IgG antibodies were mildly positive. We conclude that the presence of multiple genetic and environmental risk factors greatly amplifies the risk of clinical thrombotic events. PMID- 18388508 TI - Afibrinogenemia resulting from homozygous nonsense mutation in A alpha chain gene associated with multiple thrombotic episodes. AB - Congenital afibrinogenemia is a rare disorder characterized by the absence in circulating fibrinogen, a hexamer composed of two sets of three polypeptides (Aalpha, Bbeta and gamma). Although predisposition to thrombosis is a well known feature of dysfibrinogenemia, the relatively frequent thrombotic manifestations seen in congenital afibrinogenemia are puzzling. We herein report a mutational analysis of a young afibrinogenemic man from Turkey with multiple thrombo-embolic events involving both arteries and veins. Purified DNAs of the propositus was used for amplification by polymerase chain reaction of all the exons of the A subunit gene with primers allowing the analysis of the intron-exon boundaries. Analysis of the genes coding for the three fibrinogen chains of the propositus found a homozygous G to A transition in the exon 5 of the A alpha chain gene (g.g4277a; access number gi458553). The TGG to TGA codon change predicts a homozygous W315X in the A alpha chain (p.W334X when referring to the translation initiation codon). Both parents and his brother were found to carry this heterozygous mutation. This is the first report of a patient homozygous for this rare mutation associated with afibrinogenemia. Our patient was free of known risk factors as well as diseases associated with thrombosis including atherosclerosis, vasculitis, Buerger's disease, and it seems therefore probable that afibrinogenemia itself might have contributed to both arterial and venous thrombosis. PMID- 18388511 TI - 'Life cycle in modern rheumatology' matters: new drugs require new therapeutic strategies and innovative diagnostics. PMID- 18388509 TI - Evaluation of the changes in hemostatic parameters induced by etoricoxib in rat model. AB - The effects of etoricoxib on blood coagulation parameters were evaluated, along with acetylsalicylic acid, in male Wistar rats. Blood samples were collected at the end of the first, second, third and fourth weeks of the administration period and the plasma concentrations of etoricoxib determined by liquid chromatography tandem mass spectrometry; the samples were also used for the analysis of the hematological parameters. There were no significant changes in the platelet count and fibrinogen levels, and a decrease by 1.9% of the prothrombin time was detected at the third week. The activated partial thromboplastin time assay showed a nonsignificant shortening. The antiactivated factor X and antiactivated factor II activities showed reductions lower than 2.8 and 1.3%, respectively. These findings and the comparison with acetylsalicylic acid can be helpful to support the benefit-to-risk profile, contributing to the safe therapeutic use. PMID- 18388512 TI - Update on the use of analgesics versus nonsteroidal anti-inflammatory drugs in rheumatic disorders: risks and benefits. AB - PURPOSE OF REVIEW: In the last 2 years, there have been numerous publications on the safety of nonsteroidal anti-inflammatory drugs and cyclo-oxygenase-2 inhibitors. An evaluation of the potential risks and benefits of other analgesics has also followed. In this time of greater analysis of analgesic use, this review seeks to present the most recent evidence. RECENT FINDINGS: Concerns of potential hepatotoxicity of therapeutic doses of paracetamol have been highlighted in the last 18 months. The efficacy and risks of long-term opioid use have also been reevaluated. The debate over nonsteroidal anti-inflammatory drug and cyclo oxygenase-2 inhibitor safety continues. SUMMARY: Recent evidence has prompted a reassessment of the safety of paracetamol in certain groups of patients. Further clarification on the risks of nonsteroidal anti-inflammatory drug and cyclo oxygenase-2 therapy for individuals is covered. Their use, increased cardiovascular risk and long-term implications need to be evaluated. PMID- 18388513 TI - Pain management in fibromyalgia. AB - PURPOSE OF REVIEW: Pain is the primary presenting symptom in the vast majority of inflammatory and noninflammatory rheumatic diseases. Patients tell us that improved pain relief is a principal concern. Many pain complaints respond incompletely to the treatment of the primary rheumatic disorder and pain syndromes such as fibromyalgia do not respond to traditional analgesic medications. Therefore, proper management requires consideration of additional medications for symptomatic relief. This review addresses newer strategies for the treatment of pain in patients with fibromyalgia that may be also useful in patients with other rheumatic diseases. RECENT FINDINGS: New medications have been developed with a better understanding of chronic pain mechanisms that principally address pain neurobiology at the levels of the spinal cord and the brain. Clinical studies demonstrate the effectiveness of the alpha-2-delta ligands (gabapentin and pregabalin) and the norepinephrine/serotonin reuptake inhibitors (duloxetine and milnacipran) in fibromyalgia. SUMMARY: Patients with chronic pain, best classified as fibromyalgia, either primary or in association with other rheumatic disorders, may experience benefit from new therapies targeting central pain mechanisms. PMID- 18388514 TI - Disease-modifying antirheumatic drugs other than methotrexate in rheumatoid arthritis and seronegative arthritis. AB - PURPOSE OF REVIEW: To outline recent research findings with nonmethotrexate disease-modifying antirheumatic drugs in rheumatoid arthritis and seronegative arthritis spanning systematic reviews, randomized controlled trials, observational clinical practice trials and assessments of adverse effects. RECENT FINDINGS: Systematic reviews show no important differences between methotrexate, leflunomide and sulfasalazine monotherapies; early disease-modifying antirheumatic drug therapy reduces erosive progression. Observational studies show that nonmethotrexate disease-modifying antirheumatic drugs are widely prescribed; their usage has increased in the biologic era. A systemic review also showed patients who failed monotherapy benefited from disease-modifying antirheumatic drug combinations without excess toxicity. Randomized controlled trials of intensive initial disease-modifying antirheumatic drug combinations showed they reduce synovitis and erosive damage, especially when used with steroids. The subsequent sequence of disease-modifying antirheumatic drugs and the value of changing disease-modifying antirheumatic drug monotherapies or stepping-up to combination disease-modifying antirheumatic drugs are, however, unresolved. The adverse risks of nonmethotrexate disease-modifying antirheumatic drugs have been evaluated, including infections and lung disease; patient-related risks seem more important than drug-related risks, though several disease modifying antirheumatic drugs increase both types of adverse reactions. Two limitations of nonmethotrexate disease-modifying antirheumatic drugs are reduced impact on comorbidities like cardiovascular disease and reduced patient and clinician preferences for these treatments. SUMMARY: Nonmethotrexate disease modifying antirheumatic drugs are effective, relatively well tolerated and widely used. Their role in intensive treatment strategies in early rheumatoid arthritis appears of crucial importance. PMID- 18388515 TI - Is there a future for small molecule drugs in the treatment of rheumatic diseases? AB - PURPOSE OF REVIEW: In this review, we outline the landscape of recent developments regarding small molecule compounds for the treatment of inflammatory disorders by discussing drug candidates currently in the pipeline. We also stress the fact that novel techniques are available to evaluate the safety of new therapeutics at an early stage of development. RECENT FINDINGS: Regulation of signal transduction has evolved into an important field of drug research, and small molecule inhibitors of a number of pathways are tested as new anti inflammatory agents. For rheumatic diseases, specific Jak3 and Syk inhibitors are, so far, the most successful compounds due to their good efficacy, representing a significant advantage over p38 mitogen-activated protein kinase inhibitors. Additional benefit in the treatment of inflammatory diseases may be provided by targeting CD80, IL-12/IL-23, AP-1 transcription factor and receptors modulating cellular activation like chemokine receptors, Toll-like receptors and adenosine A3 receptor. SUMMARY: There is a big hope that novel small molecule drugs, which are rationally designed, based on scientific advancements and biotechnological improvements, will achieve or even exceed efficacy of protein drugs. Thereby, new therapeutic alternatives would be given, and chances for improved outcomes in the care of rheumatic patients provided. PMID- 18388516 TI - New approaches of B-cell-directed therapy: beyond rituximab. AB - PURPOSE OF REVIEW: This study reviews therapeutic approaches of direct and indirect B-cell targeting in autoimmune diseases and their impact on protective immunity. RECENT FINDINGS: Beyond recent clinical experiences with rituximab as B cell-depleting agent, other biologicals targeting CD20, such as ocrelizumab, ofatumumab, hA20, and TRU-015 mainly deplete B cells and are under clinical investigation in different entities. Moreover, anti-CD22 targeting as another approach that has been studied in clinical trials showed a modest depletion, but inhibition of B-cell activation. More indirect innovative B-cell-affecting therapies comprise blockade of cytokines, such as B-cell-activating factor (BAFF/BLyS), APRIL, and their receptors as well as blockade of costimulation. Although decreases of immunoglobulin levels were seen, so far no major increases in infections were reported. SUMMARY: The value of certain B-cell-depletion therapies as well as other therapies modulating B-cell functions needs to be further delineated, especially in the therapeutic regimen of rheumatoid arthritis, specific collagen vascular diseases and vasculitis. Long-term observations of protective immunity are also needed to further evaluate the rate of infections. PMID- 18388517 TI - At the horizon of innovative therapy in rheumatology: new biologic agents. AB - PURPOSE OF REVIEW: To review the rational and the results regarding the use of novel biologic agents in inflammatory rheumatic diseases. RECENT FINDINGS: Recent findings show that excessive IL-1 processing and release contribute to different rheumatic conditions, including periodic fever syndromes, systemic-onset juvenile idiopathic arthritis, adult Still's disease, and crystal-induced arthritis. Preliminary results indicate that administration of IL-1 receptor antagonist and other IL-1 inhibitors improves these conditions. IL-6 also plays a major role in the control of inflammatory responses. Several clinical trials have shown that inhibition of IL-6 by a monoclonal antibody against its receptor is efficacious in rheumatoid arthritis and systemic-onset juvenile idiopathic arthritis patients. Accumulating evidence indicates that other cytokines, including IL-15, IL-18, and IL-21 may also play an important role in rheumatoid arthritis. Several signaling pathways involved in the immune and inflammatory responses may also constitute novel targets. Preliminary data on an agent targeting the Janus kinase/Signal transducer and activators of transcription pathway are encouraging. SUMMARY: Beyond tumor necrosis factor alpha targeting, the use of inhibitors against other cytokines and cytokine-induced intracellular responses is leading to a promising therapy in the future. PMID- 18388518 TI - Management of chronic low back pain in 2007-2008. AB - PURPOSE OF REVIEW: To report and critically appraise the current state of managing chronic (and subacute) low back pain as reflected in recently published guidelines and results from latest trials. RECENT FINDINGS: Although occurrence rates of and social benefits for back pain are apparently on the decline (at least in Germany), it is still one of the most frequent and costly health disorders in Western countries. Ten percent of the adult German population suffers from chronic disabling back pain. For this group, a plethora of interventions is recommended. The most recent guideline from the US alone enumerates eight different treatments based on 'moderate-quality evidence'. Latest (often high-quality) trials from 2006 to 2007 strengthen the evidence base, only modestly, owing to their conflicting and/or confusing results. SUMMARY: Recommendations for the management of chronic disabling nonspecific back pain have made little progress over the past 10 years. The field still suffers from a 'strong weakness' due to a generally weak body of evidence. Progress can be expected when the effects of an intervention can be reliably calibrated against comparators of known (in)effectiveness. At present, an individualized clinical approach seems most appropriate. Patients and doctors are free to choose, according to their experiences and preferences, from recommended treatments. PMID- 18388519 TI - Does physical therapy still have a place in the treatment of ankylosing spondylitis? AB - PURPOSE OF REVIEW: To review studies of various physical therapy programs in ankylosing spondylitis and identify their benefits and potential indications in the treatment of this disease. RECENT FINDINGS: Various exercise and physical therapy programs have been evaluated in clinical studies. Home exercise programs have been shown to improve symptoms, mobility, function and overall quality of life. Formal physical therapy under the supervision of a physical therapist has been shown to improve posture, fitness, mobility, function and mood. Water therapy may improve symptoms, function and overall sense of health. Inpatient rehabilitation may provide rapid short-term improvement in pain and stiffness, mobility, function and quality of life for patients with severe active disease. SUMMARY: Despite the advances in the pharmacological therapy of ankylosing spondylitis, physical therapy remains an essential part of the management plan. Even though data are not sufficient to determine which specific physical therapy program should be recommended, physicians should implement such nonpharmacological therapy as part of a comprehensive management strategy for this disease. All patients should receive instructions on proper posture and home exercises and be encouraged to perform water exercises if they can. Formal physical therapy and, in most severe cases, inpatient rehabilitation may be of benefit to select patients with ankylosing spondylitis. PMID- 18388520 TI - Innovative treatment strategies for patients with rheumatoid arthritis. AB - PURPOSE OF REVIEW: The present review provides an update on novel treatment strategies striving for remission in patients with recent onset of rheumatoid arthritis. RECENT FINDINGS: As early treatment is crucial to achieve optimal results, identifying patients with rheumatoid arthritis early is imperative to achieve clinical remission. Patients with early arthritis who will progress to rheumatoid arthritis can be identified, and treating these patients can postpone the diagnosis of rheumatoid arthritis and retard the progression of structural damage. The best way to achieve remission is by adjusting treatments at regular intervals using predetermined response criteria. Specific treatments to rapidly induce remission include disease modifying antirheumatic drugs combinations, especially combined with glucocorticoids or tumor necrosis factor antagonists. The prediction of joint damage progression, or the response to specific drugs is not yet accurately possible. The early institution of tumor necrosis factor antagonists followed by discontinuation leads to sustained clinical benefit. SUMMARY: Early treatment of patients with rheumatoid arthritis with strategies aiming at remission results in the best outcomes. Until the prediction of a severe disease course and treatment response becomes possible, a promising strategy would be to rapidly induce remission using an effective combination of drugs followed by tapering and discontinuation. Tumor necrosis factor antagonists have proven to be highly effective in this approach. PMID- 18388521 TI - Immunization of patients with rheumatoid arthritis with antitumor necrosis factor alpha therapy and methotrexate. AB - PURPOSE OF REVIEW: The aim of this study is to highlight the recent findings on the use of methotrexate and/or TNFalpha-blockers in adult patients with rheumatoid arthritis and their effects on the immune response to various vaccines. RECENT FINDINGS: Regarding influenza vaccination, methotrexate monotherapy is not associated with a decreased response, whereas the use of etanercept and infliximab in combination with methotrexate may cause lower titers and lower response rates. Concerning pneumococcal vaccination, methotrexate seems to impair responsiveness. The concomitant use of adalimumab and methotrexate is also associated with decreased response, whereas the concomitant use of etanercept or infliximab seems not to have an effect on response rates. As immunological pathways seem to play a major role, T-cell-dependent pneumococcal vaccines are designed to achieve higher response rates and protective titers. SUMMARY: Patients with rheumatic disorders are more likely to develop preventable infectious diseases, which underlines the importance of adequate immunoprotective titers. Several studies have shown that the combination of methotrexate and certain TNFalpha-blockers are affecting the responsiveness to vaccines. Further findings indicate that the response also depends on what type of vaccine is used. PMID- 18388522 TI - Antibodies to citrullinated proteins in arthritis: pathology and promise. AB - PURPOSE OF REVIEW: The purpose of this review is to describe how the current knowledge of antibodies to citrullinated protein antigens in rheumatoid arthritis and related conditions emerged; to discuss the diagnostic and prognostic value associated with antibodies to citrullinated protein antigens as a biomarker; and most importantly for this review, to discuss the potential pathogenetic significance of these antibodies. RECENT FINDINGS: Antibodies to citrullinated protein antigens have evolved from being mainly a diagnostic marker, to being recognized as something that can help us understand fundamental etiologic and pathogenetic features of rheumatoid arthritis. Fundamental in this context is the finding that rheumatoid arthritis can be divided into two distinct subsets by means of presence or absence of antibodies to citrullinated protein antigens. Thus, several genetic as well as environmental risk factors differ between these two variants of rheumatoid arthritis. From analysis of these genetic and environmental risk factors, new testable hypotheses have been produced concerning triggering of antibodies to citrullinated protein antigens, and potential pathogenicity of antibodies to citrullinated protein antigens and accompanying immune reactions. SUMMARY: The implications of the findings are that antibodies to citrullinated protein antigens can be used for early and precise diagnosis of a subset of rheumatoid arthritis with worse prognosis than other polyarthritides, and that a new basis is formed for etiologic and pathogenetic studies of antibodies to citrullinated protein antigens-positive rheumatoid arthritis. PMID- 18388523 TI - Activity assessments in rheumatoid arthritis. AB - PURPOSE OF REVIEW: To describe the importance of assessing disease activity in general, aiming for significant improvement particularly low disease activity and remission, and the value of employing simplified instruments toward this end in rheumatoid arthritis RECENT FINDINGS: Various instruments have either been newly developed, validated or assessed in the recent two years. Additional insights relate to the frequency of attainment of low disease activity and remission in clinical trials and clinical practice, as well as to therapeutic strategies, which involve comprehensive and tight evaluation of disease activity in the adaptation of therapy, including biologicals. All studies assessing these instruments reveal that simplified scores perform at least similar compared with more complex indices, and often better, in the evaluation of disease activity and response to treatment. SUMMARY: Simplified indices can be routinely used in clinical practice and trials and adaptation of therapy on the basis of tight control of disease activity will lead to improved outcomes of rheumatoid arthritis. PMID- 18388524 TI - Biologic therapy for early rheumatoid arthritis: the latest evidence. AB - PURPOSE OF REVIEW: To describe current therapeutic trials with biologic agents for early rheumatoid arthritis, analyzing clinical and radiographic outcomes. RECENT FINDINGS: The use of tumor necrosis factor-alpha inhibitors in combination with disease-modifying antirheumatic drugs early after the diagnosis of aggressive rheumatoid arthritis seems to provide increased clinical benefit over methotrexate or tumor necrosis factor-alpha inhibitors as monotherapy, with better outcomes in terms of faster and more extensive clinical improvement. There also seems to be an increased likelihood of low-disease activity in some cases even after tapering therapy. Control of radiographic progression appears to be most effective among early rheumatoid arthritis patients treated with combination tumor necrosis factor-alpha inhibitors and methotrexate, although radiographic outcomes are better with tumor necrosis factor-alpha inhibitor monotherapy than with methotrexate alone. SUMMARY: The addition of antitumor necrosis factor-alpha agents to traditional disease-modifying antirheumatic drugs in early rheumatoid arthritis is a novel strategy which follows the principle of early and aggressive therapeutic intervention. Results from recent trials show greater levels of disease control. The impact on long-term safety and cost-efficacy are factors which will need to be better characterized over time. PMID- 18388525 TI - Association of infections and tuberculosis with antitumor necrosis factor alpha therapy. AB - PURPOSE OF REVIEW: Rheumatoid arthritis patients have higher risk for infections due to comorbidities, underlying immunosuppression and use of glucocorticoids and disease modifying antirheumatic drugs. The association between treatment with antitumor necrosis factor alpha agents and serious infections, including opportunistic infections such as tuberculosis, in rheumatoid arthritis patients remains controversial. We present recent literature on this topic with a focus on clinical applications of this new data. RECENT FINDINGS: Prospective cohort studies and population-based registries have described the incidence and risk of serious infections in large rheumatoid arthritis patient populations of antitumor necrosis factor alpha users. Although some studies have suggested a one and one half to two-fold increased risk, especially immediately after initiating the treatment, not all have shown an elevated risk for serious bacterial infections or tuberculosis. SUMMARY: Although antitumor necrosis factor alpha agents may be independent risk factors for infections there is an absolute low rate of infection in those treated with these agents (approximately 5 per 100 patient years). Screening for latent tuberculosis with tuberculin skin testing is effective, and compliance with the recommendations for preventing this disease in recipients of antitumor necrosis factor alpha agents has partially decreased the risk of infections. Clinical suspicion toward developing infection in those being treated with antitumor necrosis factor alpha agents, particularly earlier in the treatment course, is important for effective management of patients. PMID- 18388527 TI - Malignancy and biologic therapy in rheumatoid arthritis. AB - PURPOSE OF REVIEW: Owing to the complex functions of the inflammatory response systems--potentially or clearly of importance in human carcinogenesis--that biological therapies interfere with uncertainty regarding their safety profile for malignancy is more or less expected. This uncertainty has been further sparked by the apparent discordance between trial data and observational studies of anti-TNF agents, and the methodological challenges inherent in addressing the safety profile of new drugs for delayed and multifactorial events like cancer. RECENT FINDINGS: This review provides a summary of the pattern of cancer seen in patients with rheumatoid arthritis not treated with biologics, and the currently published data on cancer risk following treatment with biologics in patients with rheumatoid arthritis, primarily anti-TNF therapy. SUMMARY: Published data currently do not exclude clinically important increased risks, nor do they refute beneficial effects. As per definition, much of the currently available safety data from trials or clinical practice do not capture the impact of either any effect that biological therapy might have on early events in carcinogenesis, or of sustained exposure to biologics. Beyond the risk of de-novo cancer development, several other clinically important aspects of cancer safety remain to be addressed, including issues of prognosis, progression, and relapse. PMID- 18388526 TI - Biologics and heart failure in rheumatoid arthritis: are we any wiser? AB - PURPOSE OF REVIEW: To summarize the recent literature concerning the role of TNF alpha in heart failure, epidemiology of heart failure in rheumatoid arthritis and risk of heart failure associated with biologic disease-modifying antirheumatic drugs in rheumatoid arthritis. RECENT FINDINGS: TNF-alpha has been implicated in the pathogenesis of heart failure. It has direct deleterious effects on the myocardium in the setting of acute injury or chronic heart failure. In animal models, TNF-alpha is important in cardiac remodeling, leading to cardiac dysfunction following acute injury. Both incident and worsening heart failure have been reported in patients with rheumatoid arthritis who are treated with anti-TNF-alpha therapy. Recent cohort studies, however, have shown no increased risk and, in some, a protective effect on the risk of heart failure. Certain traditional cardiovascular risk factors have a relatively lesser contribution to cardiovascular morbidity and mortality in patients with rheumatoid arthritis, suggesting that disease-related perturbations of the cytokine network may contribute to the excess risk of heart failure in these patients. SUMMARY: Overall mortality in rheumatoid arthritis has remained stagnant despite advances in rheumatoid arthritis and heart failure management and improved cardiovascular mortality in the general population. Heart failure prevalence is increased in patients with rheumatoid arthritis and leads to greater mortality. Despite current expert consensus contraindicating the use of anti-TNF-alpha agents in patients with moderate to severe heart failure, epidemiological studies in rheumatoid arthritis have not consistently substantiated this association. PMID- 18388528 TI - Lung disease in rheumatoid arthritis. AB - PURPOSE OF REVIEW: To examine the role of lung disease in rheumatoid arthritis from a clinical, epidemiologic, pathophysiologic, and therapeutic perspective. RECENT FINDINGS: Lung disease in rheumatoid arthritis is pleomorphic and has a marked adverse impact on the morbidity and premature mortality of patients with this disease. Recent advances in the understanding of the pathophysiology of lung disease associated with rheumatoid arthritis reveal it to be characterized by more active cellular infiltrates of both T cells and B cells, as well as other immunologically active cells, including mast cells, than many of the other forms of interstitial lung disease. Satisfactory treatment is lacking; available biologic response modifiers have been reported to have both beneficial and adverse effects on the lung. Newer approaches targeting cellular immunologic dysfunction including T-cell-directed and B-cell-directed therapies hold the promise of reducing lung damage related to the underlying disease. SUMMARY: Lung disease in rheumatoid arthritis is a heterogeneous and oftentimes serious condition, with a profound impact on patient wellbeing and survival. Advances in the understanding of its etiology and targeted application of available, as well as development of new, more specific therapeutics will be of benefit to patients with rheumatoid arthritis who are suffering from lung disease. PMID- 18388529 TI - Vasculitis in Behcet's syndrome: evidence-based review. PMID- 18388530 TI - Exercise therapy and orthotic devices in rheumatoid arthritis: evidence-based review. PMID- 18388531 TI - Bibliography. Current world literature. Clinical therapeutics. PMID- 18388532 TI - Healthcare law, ethics, and regulation. PMID- 18388533 TI - Legal briefs. PMID- 18388534 TI - Conversations in ethics. PMID- 18388535 TI - Advanced practice registered nurse educational programs and regulation: a need for increased communication. AB - Communication is essential between nursing education program and regulation to ensure that graduates are prepared to meet licensure eligibility requirements. Not only should educational programs be aware of their own state requirements but they should also be knowledgeable of the variations of eligibility requirements among the states. Formal and informal meetings between educators and regulators are encouraged. PMID- 18388536 TI - To hold her hand: family presence during patient resuscitation. AB - Family presence at the bedside during resuscitation is an important component of the patient's care. Many families report feeling that their presence at such a time is helpful to both them and the patient. Some studies suggest that family presence may reduce the chance of legal action regarding the patient's outcome because it decreases the mystery surrounding the level of effort undertaken to save the patient's life. However, many facilities are reluctant to allow family presence during resuscitation typically because of the belief that family presence will somehow disrupt the providers' ability to conduct the resuscitation. This article explores the background behind this issue and the studies done to date on family presence and makes suggestions for adopting policies allowing family presence during resuscitation. PMID- 18388537 TI - Do psychiatric advance directives protect autonomy? AB - A psychiatric advance directive (PAD) is a legal resource that allows a person to document his or her wishes about receiving mental health services in the event that he or she is unable to provide consent at a future time. Advantages for completing a PAD include promoting the person's autonomy and authentic self, minimizing harms, and promoting beneficence. However, more research is needed to describe the positive and negative consequences associated with completing and implementing a PAD in the clinical setting. PMID- 18388540 TI - Surgery of the human cerebrum. Endoscopy. Selected bibliography. PMID- 18388539 TI - The case manager as expert witness. PMID- 18388541 TI - Surgery of the human cerebrum. Infections and infestations. Selected bibliography. PMID- 18388542 TI - Surgery of the human cerebrum. Epilepsy. Selected bibliography. PMID- 18388544 TI - How secure is your practice? PMID- 18388543 TI - Teamwork results in new practice standards. PMID- 18388545 TI - Restless leg syndrome: iron important. PMID- 18388547 TI - Treatment avenues for diabetic neuropathy. PMID- 18388548 TI - Current acceptance of the HPV vaccine. PMID- 18388550 TI - Little arms, big league injuries. PMID- 18388552 TI - ADHD treatment and the risk of substance abuse. PMID- 18388553 TI - Exposing barriers to asthma care in Hispanic children. PMID- 18388554 TI - New techniques in carotid stenting. PMID- 18388556 TI - Gait analysis on split-belt force treadmills: validation of an instrument. AB - OBJECTIVE: Gait analysis (GA) is usually performed during overground walking, with ground reactions being recorded through force platforms embedded in the floor. Body kinematics and surface electromyography (EMG) are also recorded. GA on treadmills resting on force sensors (GAFT) is rather uncommon and has been criticized on various grounds (less reliability of force records; biomechanical differences in walking kinematics and dynamics). On the other hand, the treadmill allows very fast recording of data from many successive strides, at known and constant average speeds. The goals of this study were (a) to validate a system for GAFT based on a commercially available "split-belt" force treadmill, (b) to provide evidence that data can be compared with established norms from overground GA, and (c) to lend support to the potential clinical validity of the GAFT approach. DESIGN: The treadmill adopted here allows subjects to walk on two parallel independent treadmills. Each of them is mounted on four 3D force sensors. Eight healthy adults (four women; ages 22-35 yrs) were tested on the treadmill, each at the average walking speed he or she adopted overground (average across subjects: 1.35 +/- 0.05 m x sec(-1)). Ground reactions, hip, knee, and ankle sagittal rotations, torques, power, and surface EMG from four thigh and leg muscles were recorded simultaneously. RESULTS: Results were compared with those relating to a sample of 40 healthy adults (20 women; manufacturer's data, replicating published data) walking on floor-embedded force platforms (age 20-40 yrs; speed 1.33 +/- 0.06 m x sec(-1)). An 8% shorter stride length was adopted on the treadmill. All of the other results matched those obtainable overground. CONCLUSIONS: The results suggest that GAFT performed on the tested treadmill is a promising method of GA in a clinical setting. PMID- 18388557 TI - Amplitude ratio of ulnar sensory nerve action potentials in segmental conduction study: reference values in healthy subjects and diagnostic usefulness in patients with ulnar neuropathy at the elbow. AB - OBJECTIVE: To determine normal values for the amplitude ratio of sensory nerve action potential (SNAP) from an ulnar sensory segmental nerve conduction study, and to the evaluate usefulness in the diagnosis of mild-degree ulnar neuropathy at the elbow (UNE). DESIGN: Segmental sensory conduction study of the ulnar nerve was performed in 71 healthy subjects. Peak latency and baseline to peak amplitudes were measured. The amplitude ratio of below-elbow to wrist (BE/W) stimulations and above-elbow (AE) to below-elbow stimulations (BE) were calculated. Normal cutoff values were obtained by subtracting 2 SD from the mean value, and these values were applied to 22 symptomatic UNE cases. The amplitude ratio was also obtained in six subjects with C8 radiculopathy. RESULTS: The amplitude ratios of BE/W and AE/BE were 0.61 +/- 0.08 and 0.82 +/- 0.08, respectively. The cutoff value of BE/W was 0.45, and that of AE/BE was 0.65. The amplitude ratio of BE/W showed a weak correlation to the length of the forearm segment (r = -0.25, P < 0.05). Five of the 22 UNE patients revealed only reduced amplitude ratios of SNAP across the lesion, whereas all the patients with C8 radiculopathy showed normal amplitude ratios of ulnar SNAP. CONCLUSIONS: The amplitude ratio of ulnar SNAPs may be useful in the diagnosis of mild ulnar neuropathy with only sensory symptoms and normal segmental motor conduction. PMID- 18388558 TI - Electrodiagnostic medicine skills competency in physical medicine and rehabilitation residents: a method for development and assessment. AB - This project sought to create an educational module including evaluation methodology to instruct physical medicine and rehabilitation (PM&R) residents in electrodiagnostic evaluation of patients with neuromuscular problems, and to verify acquired competencies in those electrodiagnostic skills through objective evaluation methodology. Sixteen residents were trained by board-certified neuromuscular and electrodiagnostic medicine physicians through technical training, lectures, and review of self-assessment examination (SAE) concepts from the American Academy of Physical Medicine & Rehabilitation syllabus provided in the Archives of Physical Medicine and Rehabilitation. After delivery of the educational module, knowledge acquisition and skill attainment were measured in (1) clinical skill in diagnostic procedures via a procedure checklist, (2) diagnosis and ability to design a patient-care management plan via chart simulated recall (CSR) exams, (3) physician/patient interaction via patient surveys, (4) physician/staff interaction via 360-degree global ratings, and (5) ability to write a comprehensive patient-care report and to document a patient care management plan in accordance with Medicare guidelines via written patient reports. Assessment tools developed for this program address the basic competencies outlined by the Accreditation Council for Graduate Medical Education (ACGME). To test the success of the standardized educational module, data were collected on an ongoing basis. Objective measures compared resident SAE scores in electrodiagnostics (EDX) before and after institution of the comprehensive EDX competency module in a PM&R residency program. Fifteen of 16 residents (94%) successfully demonstrated proficiency in every segment of the evaluation element of the educational module by the end of their PGY-4 electrodiagnostic rotation. The resident who did not initially pass underwent remedial coursework and passed on the second attempt. Furthermore, the residents' proficiency as demonstrated by the evaluation after implementation of the standardized educational module positively correlated to an increase in resident SAE scores in EDX compared with resident scores before implementation of the educational module. Resident proficiency in EDX medicine skills and knowledge was objectively verified after completion of the standardized educational module. Validation of the assessment tools is evidenced by collected data correlating with significantly improved SAE scores and American Association of Neuromuscular and Electrodiagnostic Medicine (AANEM) exam scores, as outlined in the result section. In addition, the clinical development tool (procedure checklist) was validated by residents being individually observed performing skills and deemed competent by an AANEM certified physician. The standardized educational module and evaluation methodology provide a potential framework for the definition of baseline competency in the clinical skill area of EDX. PMID- 18388559 TI - Internal iliac artery pseudoaneurysm: an unusual cause of sciatica and lumbosacral plexopathy. AB - This report describes an unusual case of lumbosacral plexopathy resulting from internal iliac artery pseudoaneurysm. A 50-yr-old woman presented with multiple penetrating trauma to the thorax, abdomen, and left buttock. Several weeks after the injury, severe sciatica and motor dysfunction developed in her left leg. Progressively worsening pain was followed by left foot drop. An electrodiagnostic evaluation suggested a lower lumbosacral plexopathy. Magnetic resonance imaging showed a hematoma extending adjacent to the left lumbosacral plexus. Computerized tomographic angiography revealed a left internal iliac artery pseudoaneurysm. The pseudoaneurysm was excised surgically. In the early postoperative period, the patient reported total relief of pain. Her motor function recovered gradually over several weeks; however, some residual weakness of foot dorsiflexion persisted. With this case presentation, we underscore that an arterial pseudoaneurysm should be remembered as an etiologic possibility of lumbosacral plexopathy and sciatica, especially in patients with history of iatrogenic or accidental trauma. An increased awareness of this rare cause of sciatica and lumbosacral plexopathy may enable early intervention alternatives. PMID- 18388560 TI - Ten-year epidemiological study of pediatric burns in Canada. AB - The aim of this study was to report on the temporal trends, incidence rates, demographic, and external-cause data for all burn injury related deaths and hospital admissions among children Canadian aged 0 to 19 years for the years 1994 to 2003. Statistics Canada and Canadian Institute of Health Information data were used to describe burn injury related deaths and hospital admission trends in children aged 0 to 19 years who were residents of Canada (1994-2003). Population estimates were derived from census data provided by Statistics Canada. During the 10-year period, 494 children died and 10,229 were admitted to a Canadian hospital because of a burn-related injury. Males and children aged less than 5 years of age were at the highest risk of injury, with children aged 1 to 5 years at the highest risk of death. Scalds represented the major etiological factor contributing to thermal injuries accounting for 50% of all hospital admissions. Temporal trends indicate a significant a significant decline in burn injuries across all age groups during the period 1994 to 2003. There has been a clear reduction in the number of patients with burn injury requiring hospital admission. This trend indicates success in safety initiative to prevent burn injuries as well as in improvements in the treatments of burn and hospital admission procedures. Nonetheless, burn injury remains a serious threat to the well-being of the Canadian pediatric population. PMID- 18388561 TI - Important considerations for the burn therapist. PMID- 18388562 TI - Ecthyma gangrenosum following toxic epidermal necrolysis syndrome in a 3-year-old boy-a survivable series of events. AB - Toxic epidermal necrolysis syndrome (TENS) is a severe but rare skin reaction leading to epidermal desquamation of greater than 30% of the TBSA. It is most commonly precipitated by the administration of medication. Frequent complications of this syndrome include local wound infections, respiratory, mucocutaneous, and ocular complications. Ecthyma gangrenosum (EG) is a rare disease characterized by a milliary seeding of the cutaneous tissue with Gram-negative bacteria; it is most commonly seen in immunocompromised individuals. Here we report a 3-year-old boy who developed EG subsequent to TENS. Although he had a complicated and prolonged hospital course, he survived these series of events. To our knowledge, this is the first reported case of TENS/EG in the pediatric population, and the first report of survivability following these illnesses. PMID- 18388563 TI - Carbon monoxide-induced status epilepticus in an adult. PMID- 18388564 TI - Self-immolation in Iran. AB - Deliberate self-inflicted burn is rare in high-income countries, but is reported more frequently in low- and middle-income countries, especially in Asia and Africa. Rates in Iran are among the highest in the world, with up to 71% of committed suicides conducted via self-immolation in some regions. The objective of this study was to identify the epidemiologic features and factors of self immolation in Iran to aid in development of effective intervention programs. In a review study, two national databases were analyzed to identify demographic, geographic, cultural, economic, and health-related aspects of self-immolation that may vary across regions of Iran. Demographic information revealed that 27% of suicide cases in Iran were via self-immolation. Of those, 71% were female. The mean age was 29 years. Geographical features of self-immolation indicated that the self-immolation rate was higher in rural areas and in provinces that border the country. Provinces that were involved more intensively in postwar problems feature higher rates of self-immolation. People of Kurdish ethnicity were more likely to engage in self-immolation. Unemployment was a risk factor for self immolation, while mental disorders and lack of access to health and treatment facilities did not play an important role for increasing the rate of self immolation. Overall, this study demonstrates that self-immolation is a significant public health problem in some parts of Iran. This study clearly and consistently shows that self-immolation is a complex phenomenon with multiple causes. Various intervention options are discussed to increase coping skills on the individual and community levels. During the long-term, programs and strategies should focus on "macrosocial-based" interventions. PMID- 18388565 TI - Assault and substance abuse characterize burn injuries in homeless patients. AB - The homeless are at an increased risk for traumatic injury, but little is known about the injury etiology and outcome of homeless persons who sustain burn injuries. In this study, we analyze patient and injury characteristics of homeless persons admitted to a regional burn center. This is a retrospective cohort study of patients admitted to our burn center between 1994 and 2005. A total of 3700 adult patients were admitted during the study period and, of these, 72 (1.9%) were homeless. The cohort of homeless patients was compared with domiciled adult patients admitted during the same time period, analyzing baseline patient and injury characteristics and injury outcomes. Overall, homeless patients had more extensive burn injuries than domiciled patients (17.8% vs 11.2%TBSA, P < .001) and overall longer lengths of hospital stay (22 vs 12 days, P < .001). The homeless population also had significantly higher rates of alcohol (80.6% vs 12.8%, P < .001) and drug abuse (59.4% vs 12.8%, P < .001), history of mental illness (45.2% vs 11.0%, P < .001), and injury by assault (13.9% vs 2.0%, P < .001). Homeless patients tended to have more severe injuries; higher rates of substance abuse and mental illness; increased incidence of assault by burning; and longer lengths of hospital stay. Hospitalization of a homeless patient following injury may provide a unique opportunity to address co-occurring substance abuse and mental illness and approach injury prevention to improve patients' outcomes and reduce injury recidivism. PMID- 18388566 TI - Profound effects of burn and ethanol on proinflammatory cytokines of the reproductive axis in the male mouse. AB - Thermal injury is often associated with previous ethanol exposure, and close to 50% of patients admitted to a burn unit have a potentially high blood ethanol level. Cellular mechanisms by which ethanol and/or burn affect the hypothalamic pituitary-gonadal (HPG) axis are not entirely understood. However, it is known that the proinflammatory cytokines, tumor necrosis factor-alpha, interleukin 1beta, and interleukin-6 influence negatively on the endocrine functions of the HPG. We report a time course study (6, 12, 24, and 48 hours) of the effects of ethanol, burn, or the combination of burn/ethanol on proinflammatory cytokines of the hypothalamus, pituitary and testes of male C57Bl/6 mice. We found that there were highly significant increases in each of these cytokines caused by ethanol, burn, and burn/ethanol compared with sham/vehicle (P < .001). This was true in hypothalamus, pituitary, and testes. Because these cytokines generally reduce reproductive function, it may be that proinflammatory cytokines of HPG axis mediate the deleterious effects of burn and/or ethanol on mammalian reproduction. PMID- 18388568 TI - Invasive Vibrio cholerae infection following burn injury. AB - Vibrio cholerae is a pathogen predominantly appreciated for its potential to produce life-threatening watery diarrhea, usually without invasive disease. However, nonepidemic forms, which are present worldwide, may have a severe invasive presentation, especially among those with liver disease or other immunocompromised states. We present a case of invasive infection (pulmonary, wound, and bacteremia) by nonepidemic V. cholerae, in a soldier that sustained burn injury in Iraq. Multiple factors, to include burn injury and water exposure, likely contributed to this presentation. A brief discussion of the pertinent literature is included. PMID- 18388567 TI - Inhibition of protein tyrosine phosphatases prevents mesenteric lymph node T-cell suppression following alcohol intoxication and burn injury. AB - Previously, we have shown that acute alcohol (EtOH) intoxication before burn injury potentiates the suppression of mesenteric lymph node T-cell effector responses. Moreover, the suppression in T-cell was accompanied with a decrease in p-38 and extracellular-signal-regulated kinase (ERK) activation. This study examined the role of protein tyrosine phosphatases (PTP) in suppressed T-cell p 38, ERK, and cytokine production after EtOH intoxication and burn injury. A blood EtOH level of approximately 100 mg/dl in male rats (approximately 250 g) was achieved by gavaging animals with 5 ml of 20% EtOH suspension 4 hours before burn or sham injury (approximately 12.5% or 25% total body surface area [TBSA]). One day after injury, rats were killed and mesenteric lymph node T-cell cytokine (IL 2/IFN-gamma) production, p-38, and ERK activation were measured. As compared with shams, there was a significant decrease in T-cell cytokine production after 25% and not 12.5% TBSA burn injury. However, T-cell IL-2/IFN-gamma levels were significantly decreased in rats receiving a combined insult of EtOH and burn injury regardless of the percentage of burn area. Furthermore, we found a significant decrease in p-38 and ERK-1/2 phosphorylation in T-cells of rats receiving a combined insult of EtOH and 12.5% TBSA burn compared with shams. Treatment of cells with PTP inhibitor pervanadate (10 muM) prevented T-cell p 38/ERK suppression. The suppression in IL-2/IFN-gamma production was also attenuated in T-cells cultured in the presence of pervanadate. These findings suggest that an increase in PTP activity may contribute to T-cell suppression after EtOH intoxication and burn injury. PMID- 18388569 TI - Forked finger fillet flap for segmental extensor tendon coverage of an adjacent digit. AB - Finger fillet flaps have been used to treat a variety of complex hand deformities providing stable soft tissue coverage and preventing pathologic contractures. Fillet flaps have not been reported in the coverage of segmental extensor tendon deficit in an adjacent digit. A 20-year-old man involved in a motor vehicle crash sustaining a 30% total body surface area burn, primarily to left arm and bilateral lower extremities. In particular, his left index finger extensor mechanism was disrupted with exposure of the proximal interphalangeal (PIP) joint. In addition, the middle finger had a segmental exposure of the extensor tendon. The nonfunctional index finger was sacrificed to provide coverage, via a forked fillet finger flap, of the exposed extensor tendon at the PIP and metacarpophalangeal (MCP) joints. Total active motion of left fingers at 12 months consisted of the third digit 0 to 86 degrees at the MCP joint, 0 to 88 degrees at the PIP joint and 0 to 33 degrees at the distal interphalangeal joint. Gross grip strength improved to 26 lb at 1 year follow-up. Adequate soft tissue coverage of extensor tendons can be challenging in the traumatic hand. With this novel approach of a forked finger fillet flap we were able to provide adequate soft tissue coverage of exposed tendons and joints improving the patient's strength and active range of motion especially in the middle finger. Prevention of postburn boutonniere deformity was an additional benefit. PMID- 18388570 TI - Copper sulphate burns to the hands, a complication of traditional medicine. AB - An 11-year-old girl of Bedouin origin presented with infected burns on the dorsum of both her hands. The burns had an unusual configuration in the form of spirals, and child abuse was suspected. The family reported that the burns had been caused by a treatment, for head aches, dispensed by a traditional healer, which involved applying a blue fluid on the hands. Chemical analysis of the implicated substance identified it as copper sulfate. The local and systemic effects of copper sulfate are reviewed as well as traditional healing methods that may lead to burn injuries. When clinicians encounter bizarre patterns of injury they must consider besides the possibility of abuse also, more unusual etiologies. PMID- 18388572 TI - WITHDRAWN: The Etiology of Bacteremia or Pneumonia as a Prognostic Factor for Death in Burn Patients, After a 10-Day in Intensive Care Unit. PMID- 18388571 TI - Hypocalcemia, hypomagnesemia, and hypokalemia following hydrofluoric acid chemical injury. AB - Dermal exposure to hydrofluoric acid could potentially result in severe serum calcium and magnesium depletion induced by binding with fluoride anion. This report describes the case of a 48-year-old man who developed hypocalcemia and hypomagnesemia accompanied by hypokalemia-an interesting finding-following a chemical injury with exposure to 70% hydrofluoric acid. Successful treatment included administration of calcium gluconate and magnesium both intravenously and topically. PMID- 18388573 TI - Warding off burn injuries, warding off database fishing expeditions: the ABA burn prevention committee takes a turn with a glimmer from the national burn repository. PMID- 18388574 TI - The expectations and experiences of children attending Burn Camps: a qualitative study. AB - The use of evaluation forms has increased with the steadily growing interest in child-centered research. The purpose of this study is 2-fold: 1) to examine the benefits of attending the National Burn Camp, a therapeutic recreation program set up in Belgium for burn-injured children aimed at alleviating the impact of their burns, and 2) to evaluate the camp program from the standpoint of the children, their parents, and the staff members to make improvements to the program. Evaluation forms were gathered from 19 children (12-18 years), parents (N = 14), and staff members (N = 20) who agreed to fill out open-ended questions at different times. Evaluation data demonstrated evidence of increased self confidence, improved coping skills, developed social skills, new relationships, and a sense of achievement at having taken part in a therapeutic recreation program. Children genuinely benefit psychologically, according to the participants themselves, their parents, and staff members. Evaluation forms can provide additional and more profound explanatory information on the wishes and demands of the participants, their parents, and staff members. Moreover, they are of great importance when it comes to adapting and improving camp programs. PMID- 18388575 TI - HSP70 inhibits burn serum-induced apoptosis of cardiomyocytes via mitochondrial and membrane death receptor pathways. AB - The objective is to determine if Hsp70 is involved in the mechanism by which serum from burn rats induces apoptosis of cardiomyocytes. Cardiomyocytes in primary culture were assigned to the following groups: normal controls, normal rat serum, burn serum, burn serum plus empty vector control, and burn serum plus Hsp70 transfection. Each group, except normal controls, was cultured with the corresponding serum for 2 hours. The empty vector controls and the Hsp70 groups were transfected, respectively, with the empty vector or pcDNA3.1-Hsp70 recombinant plasmid 36 hours previously. Hsp70 protein levels were assessed by Western blot. Apoptotic cells were counted after Hoechst 33258 staining. Apoptosis of cardiomyocytes was also detected by the presence of "ladder" bands in agarose gels following electrophoresis of extracted DNA. Activation of caspase 3, -8, and -9 and Bid cleavage were assessed by Western blot. Hsp70 overexpression inhibited burn serum-induced apoptosis of cardiomyocytes; and burn serum-induced activation of caspases-3, -8, and -9; and Bid cleavage into tBid. Hsp70 inhibits burn serum-induced apoptosis by interfering with death receptor and mitochondrial pathways. PMID- 18388576 TI - Quantitative measurement of hypertrophic scar: interrater reliability and concurrent validity. AB - Research into the pathophysiology and treatment of hypertrophic scar (HSc) remains limited by the heterogeneity of scar and the imprecision with which its severity is measured. The objective of this study was to test the interrater reliability and concurrent validity of the Cutometer measurement of elasticity, the Mexameter measurement of erythema and pigmentation, and total thickness measure of the DermaScan C relative to the modified Vancouver Scar Scale (mVSS) in patient-matched normal skin, normal scar, and HSc. Three independent investigators evaluated 128 sites (severe HSc, moderate or mild HSc, donor site, and normal skin) on 32 burn survivors using all of the above measurement tools. The intraclass correlation coefficient, which was used to measure interrater reliability, reflects the inherent amount of error in the measure and is considered acceptable when it is >0.75. Interrater reliability of the totals of the height, pliability, and vascularity subscales of the mVSS fell below the acceptable limit ( congruent with0.50). The individual subscales of the mVSS fell well below the acceptable level (< or =0.3). The Cutometer reading of elasticity provided acceptable reliability (>0.89) for each study site with the exception of severe scar. Mexameter and DermaScan C reliability measurements were acceptable for all sites (>0.82). Concurrent validity correlations with the mVSS were significant except for the comparison of the mVSS pliability subscale and the Cutometer maximum deformation measure comparison in severe scar. In conclusion, the Mexameter and DermaScan C measurements of scar color and thickness of all sites, as well as the Cutometer measurement of elasticity in all but the most severe scars shows high interrater reliability. Their significant concurrent validity with the mVSS confirms that these tools are measuring the same traits as the mVSS, and in a more objective way. PMID- 18388577 TI - Quantitative measurement of hypertrophic scar: intrarater reliability, sensitivity, and specificity. AB - The comparison of scar evaluation over time requires measurement tools with acceptable intrarater reliability and the ability to discriminate skin characteristics of interest. The objective of this study was to evaluate the intrarater reliability and sensitivity and specificity of the Cutometer, the Mexameter, and the DermaScan C relative to the modified Vancouver Scar Scale (mVSS) in patient-matched normal skin, normal scar (donor sites), and hypertrophic scar (HSc). A single investigator evaluated four tissue types (severe HSc, less severe HSc, donor site, and normal skin) in 30 burn survivors with all four measurement tools. The intraclass correlation coefficient (ICC) for the Cutometer was acceptable (> or =0.75) for the maximum deformation measure for the donor site and normal skin (>0.78) but was below the acceptable range for the HSc sites and all other parameters. The ICC for the Mexameter erythema (>0.75) and melanin index (>0.89) and the DermaScan C total thickness measurement (>0.82) were acceptable for all sites. The ICC for the total of the height, pliability, and vascularity subscales of the mVSS was acceptable (0.81) for normal scar but below the acceptable range for the scar sites. The DermaScan C was clearly able to discriminate HSc from normal scar and normal skin based on the total thickness measure. The Cutometer was less discriminating but was still able to discriminate HSc from normal scar and normal skin. The Mexameter erythema index was not a good discriminator of HSc and normal scar. Receiver operating characteristic curves were generated to establish the best cutoff point for the DermaScan C total thickness and the Cutometer maximum deformation, which were 2.034 and 0.387 mm, respectively. This study showed that although the Cutometer, the DermaScan C, and the Mexameter have measurement properties that make them attractive substitutes for the mVSS, caution must be used when interpreting results since the Cutometer has a ceiling effect when measuring rigid tissue such as HSc and the Mexameter erythema index does not discriminate normal scar from HSc. PMID- 18388578 TI - Related factors in burn children. Epidemiological study of the burn unit at the "Magdalena de las Salinas" Traumatology Hospital. AB - We carried out a study of 1025 children. The most frequent cause of burns was hot liquids (77.7%). About 95.13% of the cases happened in the patients' homes. The cases of death and the most severe burns were due to accidents that happened in the kitchen and in the bathroom. About 74.8% of the patients were less than 5 years old; the riskiest factor was age: the younger the child, the greater the risk. The average burns extension was 11.9% with a minimum of 1% and maximum of 90%. Mortality was 0.87%; the patients who died were less than 3 years old. DL-50 was of 75.6%. Complications were presented in 10% and the most frequent was local infection in 71 cases. The most severe injuries were those that had taken place by immersion in hot liquids and fire. PMID- 18388579 TI - Influence of injury characteristics and payer status on burn treatment location in Washington state. AB - The provision of optimal burn care is a resource-intensive endeavor. The American Burn Association has developed criteria to help guide the decision to refer a patient to a burn center for definitive injury care. The purpose of this study was to compare the patient and injury characteristics of patients admitted to the single verified burn center in Washington State with those treated at other facilities in the state. We performed a retrospective review of all patients admitted to a hospital with a burn injury in Washington State from 1987 to 2005 using the state's discharge database (Comprehensive Hospital Abstract Reporting System). Patient and injury factors of patients admitted to the state's single verified burn center or at other hospitals were compared. Multivariate poisson regression was used to calculate the relative risk of injury and patient factors that were significantly associated with admission to the verified burn center. From 1987 to 2005, a total of 16,531 patients were admitted to a Washington State hospital after burn injury. Of these patients, 8624 (52.2%) were treated definitively at the University of Washington Burn Center. Patients treated at this verified center had larger overall burn size (7.4% vs 4.5% TBSA, P < .001), higher percent full-thickness burn (4.3% vs 1.2%, P < .001), and higher rates of inhalation injury (2.3% vs 1.5%, P = .005). Uninsured status (relative risk = 1.46, 95% confidence interval = 1.4-1.5) was also significantly associated with treatment at the verified burn center. Injury severity and payer status were both found to be independent predictors of treatment at the single verified burn center in Washington. PMID- 18388580 TI - A prospective analysis of trash, brush, and grass burning behaviors. AB - Burn injuries sustained during residential trash, brush, and grass burning cause significant morbidity and mortality in rural areas. To further prevention efforts, we surveyed individuals who incurred injuries from residential burning. Thirty-six individuals injured while burning trash, brush, or grass from June 2003 through September 2005 were asked to respond to a self-administered written survey. Injury related questions revealed that the majority of those injured were burning brush (21 of 35, 60.0%) in an open space (19 of 35, 54.2%) with the addition of accelerants (27 of 36, 75%). Survey questions regarding usual burning practices revealed almost two-thirds burned either brush or a mixture of brush and trash (23 of 36, 63.9%). Eighty percent of those who were injured desired to change their behavior (25 of 35, 80%). Approximately two-thirds would consider asking for help with burning if it were provided (22 of 34, 64.7%). Our survey shows that acceptable alternatives to burning varied depending on the material that was burned. As the majority of respondents usually burned brush or a mixture of brush and trash, an acceptable trash removal system should also include brush pickup. As residential burning continues presently, injury prevention efforts are essential and should focus on the misuse of gasoline, uniform safety standards for gasoline cans, and dissemination of safe burning practices. PMID- 18388581 TI - A clarion to recommit and reaffirm burn rehabilitation. AB - Burn rehabilitation has been a part of burn care and treatment for many years. Yet, despite of its longevity, the rehabilitation outcome of patients with severe burns is less than optimal and appears to have leveled off. Patient survival from burn injury is at an all-time high. Burn rehabilitation must progress to the point where physical outcomes parallel survival statistics in terms of improved patient well-being. This position article is a treatise on burn rehabilitation and the state of burn rehabilitation patient outcomes. It describes burn rehabilitation interventions in brief and why a need is felt to bring this issue to the forefront. The article discusses areas for change and the challenges facing burn rehabilitation. Finally, the relegation and acceptance of this responsibility are addressed. PMID- 18388582 TI - A population-based questionnaire study on the prevalence and epidemiology of burn patients in Denizli, Turkey. AB - Epidemiological investigations of burn patients help health services to identify the need for burn care and to plan burn care facilities. Various epidemiological studies on burn injury have been conducted, but they have usually covered major burn victims who required hospitalization or minor burn victims who were treated at health care centers. Few studies have included burn victims who were not admitted to any health care center. Through a population-based questionnaire, we studied the epidemiology of the entire population of burn victims, including those who did not seek medical attention. Thus, we identified the actual frequency of hospital admission and the frequency of sequelae. In total, 1068 persons from the city of Denizli, Turkey were questioned about burns in the last 10 years and the burn prevalence was found to be 12.6%. Only 33.3% of the burn victims were treated at a health care center; thus, 66.7% of the victims were treated at home with traditional burn wound care methods. Etiologic factors such as the age and sex of each victim, the cause and anatomic location of the burn, and location of the victim when burned were all investigated. The overall percentage of sequelae was 31.3%; of these, 89.5% were cosmetic problems, 7.9% were functional impairments, and 2.9% were physiological problems. The low percentage of health care center admissions and high incidence of sequelae suggest the need for professional burn care centers that can be easily reached by burn victims. PMID- 18388586 TI - Preparedness and response: a collaborative exercise. PMID- 18388587 TI - World Health Organization. PMID- 18388588 TI - Department of Health and Human Services. PMID- 18388589 TI - Department of Homeland Security. PMID- 18388590 TI - American Medical Association. PMID- 18388591 TI - American Nurses Association. PMID- 18388592 TI - American Red Cross. PMID- 18388593 TI - American Public Health Association. PMID- 18388594 TI - Pandemic and all-hazards preparedness act. PMID- 18388595 TI - Adverse event related to smallpox vaccination. PMID- 18388596 TI - Function of the global correspondent. PMID- 18388597 TI - Excess mortality in the aftermath of Hurricane Katrina: a preliminary report. AB - BACKGROUND: Reports that death notices in the Times-Picayune, the New Orleans daily newspaper, increased dramatically in 2006 prompted local health officials to determine whether death notice surveillance could serve as a valid alternative means to confirm suspicions of excess mortality requiring immediate preventive actions and intervention. METHODS: Monthly totals of death notices from the Times Picayune were used to obtain frequency and proportion of deaths from January to June 2006. To validate this methodology the authors compared 2002 to 2003 monthly death frequency and proportions between death notices and top 10 causes of death from state vital statistics. RESULTS: A significant (47%) increase in proportion of deaths was seen compared with the known baseline population. From January to June 2006, there were on average 1317 deaths notices per month for a mortality rate of 91.37 deaths per 100,000 population, compared with a 2002-2004 average of 924 deaths per month for a mortality rate of 62.17 deaths per 100,000 population. Differences between 2002 and 2003 death notices and top 10 causes of death were insignificant and had high correlation. DISCUSSION: Death notices from local daily newspaper sources may serve as an alternative source of mortality information. Problems with delayed reporting, timely analysis, and interoperability between state and local health departments may be solved by the implementation of electronic death registration. PMID- 18388598 TI - Characteristics of physician relocation following Hurricane Katrina. AB - INTRODUCTION: On August 29, 2005, Hurricane Katrina made landfall along the US Gulf Coast, resulting in the evacuation of >1.5 million people, including nearly 6000 physicians. This article examines the relocation patterns of physicians following the storm, determines the impact that the disaster had on their lives and practices, and identifies lessons learned. METHODS: An Internet-based survey was conducted among licensed physicians reporting addresses within Federal Emergency Management Agency-designated disaster zones in Louisiana and Mississippi. Descriptive data analysis was used to describe respondent characteristics. Multivariate logistic regression was performed to identify the factors associated with physician nonreturn to original practice. For those remaining relocated out of state, bivariate analysis with chi or Fisher exact test was used to determine factors associated with plans to return to original practice. RESULTS: A total of 312 eligible responses were collected. Among disaster zone respondents, 85.6% lived in Louisiana and 14.4% resided in Mississippi before the hurricane struck. By spring 2006, 75.6% (n = 236) of the respondents had returned to their original homes, whereas 24.4% (n = 76) remained displaced. Factors associated with nonreturn to original employment included family or general medicine practice (OR 0.42, 95% CI 0.17-1.04; P = .059) and severe or complete damage to the workplace (OR 0.24, 95% CI 0.13-0.42; P < .001). CONCLUSIONS: A sizeable proportion of physicians remain displaced after Hurricane Katrina, along with a lasting decrease in the number of physicians serving in the areas affected by the disaster. Programs designed to address identified physician needs in the aftermath of the storm may give confidence to displaced physicians to return. PMID- 18388599 TI - Preventing fatalities in building bombings: what can we learn from the Oklahoma City bombing? AB - BACKGROUND: Bombings are an increasing threat to the public's health. Descriptive studies of blast injuries have been published, but these injuries have not been studied using analytical epidemiological methods. This study assesses factors associated with fatality risk among individuals exposed to the 1995 Oklahoma City bombing. METHODS: Retrospective case-control analysis using multivariable logistic regression. Odds ratios (OR) of fatality are calculated among occupants of the Alfred P. Murrah Federal Building on April 19, 1995. RESULTS: Of the 348 occupants exposed, 163 (46.8%) were fatally injured. Fatality risk was greatest in the collapsed region of the building (adjusted OR 176.7, 95% confidence interval [CI] 65.9-474.2). Age > or =40 was also associated with a significantly increased risk of fatality (OR 3.7, 95% CI 1.4-9.8). Among people found in the noncollapsed region of the building, employees' status compared to a visitor's or child's status was protective (OR 0.13, 95% CI 0.01-1.3). CONCLUSIONS: Structural collapse is the most important risk factor for fatality in a building bombing. Progressive collapse may be prevented through more supportive building design. Protection of vulnerable building occupants can be improved by placement of relevant facilities in more structurally reinforced areas. Regular evacuation training of personnel and clear egress routes may also reduce fatality in a building bombing. PMID- 18388601 TI - Prevalence of and sex disparities in posttraumatic stress disorder in an internally displaced Sri Lankan population 6 months after the 2004 Tsunami. AB - BACKGROUND: When the 2004 Indian Ocean tsunami suddenly hit unsuspecting coastal populations in Sri Lanka, it inflicted unprecedented devastation including 35,000 deaths and 500,000 people displaced. Evaluating the psychological impact of this natural disaster provides valuable insights into planning interventions and disaster preparedness. METHODS: A cross-sectional survey was conducted among 264 adult males and females > or =16 years old living in temporary shelters housing tsunami survivors at 6 months. Interviewer-administered structured interviews were conducted to measure posttraumatic stress disorder (PTSD) and its risk factors. RESULTS: The participation rate was 97%. Of the subjects, 56% met criteria for symptoms of PTSD, with females at 64% and males at 42%. Females had at least twice the risk of experiencing PTSD (odds ratio [OR] 2.27, 95% confidence interval [CI] 1.37-3.76). This sex difference persisted after adjusting for age, marital status, being a parent, loss of family members, amount of social support, education level, and level of depression (OR 2.14, 95% CI 1.21 3.80). Depression was significantly associated with PTSD (OR 7.19, 95% CI 3.83 13.52). CONCLUSIONS: In this directly affected population, a majority met criteria for PTSD, indicating a significant long-term public health burden. The findings also confirm that females are at much higher risk for PTSD than males, suggesting that special mental health efforts should be targeted at women exposed to trauma. PMID- 18388603 TI - Voluntarism in the wake of Hurricane Katrina: the uniform emergency volunteer health practitioners act. PMID- 18388604 TI - Lifeboat ethics: considerations in the discharge of inpatients for the creation of hospital surge capacity. PMID- 18388605 TI - Public health disaster research: surveying the field, defining its future. PMID- 18388606 TI - Collateral damage. PMID- 18388607 TI - Is overtriage associated with increased mortality? Insights from a simulation model of mass casualty trauma care. AB - PURPOSE: To examine the relationship between overtriage and critical mortality after a mass casualty incident (MCI) using a simulation model of trauma system response. METHODS: We created a discrete event simulation model of trauma system management of MCIs involving individual patient triage and treatment. Model variables include triage performance, treatment capability, treatment time, and time-dependent mortality of critically injured patients. We model triage as a variable selection process applied to a hypothetical population of critically and noncritically injured patients. Treatment capability is represented by staffed emergency department trauma bays with associated staffed operating rooms that are recycled after each use. We estimated critical and noncritical patient treatment times and time-dependent mortality rates from the trauma literature. RESULTS: In this simulation model, overtriage, the proportion of noncritical patients among all of those labeled as critical, has a positive, negative, or variable association with critical mortality depending on its etiology (ie, related to changes in triage sensitivity or to changes in the prevalence and total number of critical patients). In all of the modeled scenarios, the ratio of critical patients to treatment capability has a greater impact on critical mortality than overtriage level or time-dependent mortality assumption. CONCLUSIONS: Increasing overtriage may have positive, negative, or mixed effects on critical mortality in this trauma system simulation model. These results, which contrast with prior analyses describing a positive linear relationship between overtriage and mortality, highlight the need for alternative metrics to describe trauma system response after MCIs. We explore using the relative number of critical patients to available and staffed treatment units, or the critical surge to capability ratio, which exhibits a consistent and nonlinear association with critical mortality in this model. PMID- 18388608 TI - Special issue to present complexities inherent in a response to a devastating mass casualty incident. PMID- 18388609 TI - Mass murder in a university setting: analysis of the medical examiner's response. AB - BACKGROUND: Seung-Hui Cho, a student at Virginia Polytechnic Institute and State University (Virginia Tech), shot and killed 33 students and faculty, including himself, on the morning of April 16, 2007. METHODS: A retrospective review of the medical examiner system response to this multiple fatality event was undertaken to identify which procedures were and were not effective. Case records, spreadsheets, telephone call logs, notes, and after-action interviews of staff were reviewed and analyzed. Recommendations were developed to improve the management of the multiple components of a high-profile multiple fatality event. RESULTS: One autopsy took place on Monday, April 16, 12 on Tuesday, April 17, and 20 on Wednesday, April 18. Pathologists archived the biopsies of major organs in formalin. Slides were made of entrance wounds that exhibited residues. Blood for alcohols was collected from victims. Blood for alcohols, acid, base, and neutral drugs was collected from Cho for analysis. CONCLUSIONS: Standard forensic pathology procedures worked and timely postmortem examinations were completed. The victim identification component of the family assistance center must be established and staffed at the time of the initial response. Public information officers need training in morgue and medicolegal death management and in ways to effectively communicate with different audiences about multiple fatality management procedures. PMID- 18388610 TI - A call for standardized training. PMID- 18388611 TI - Immediate behavioral health response to the Virginia Tech shootings. PMID- 18388612 TI - Building resistance, resilience, and recovery in the wake of school and workplace violence. AB - Incidents of school and workplace violence are rare but devastating events that can result in significant psychological consequences in communities. The majority of people in the United States will experience some type of traumatic event in their lifetime, but most of them will have no disruption or only transient disruption in functioning. They are either resistant to the development of symptoms or resilient, able to bounce back quickly. By enhancing resistance and promoting resilience, even fewer individuals may develop mental disorders. This article takes a closer look at the concepts of resistance, resilience, and recovery and the need for research on interventions that promote them, in the hope of applying the concepts and interventions to schools and the workplace. PMID- 18388613 TI - Preventing tragedy: balancing physicians' ethical obligations to patients and the public. AB - The recent shootings at Virginia Polytechnic Institute and State University (Virginia Tech) suggest that an increased reliance upon the medical community to support public health violence prevention efforts may be warranted. As physicians are called upon to support these efforts, they must effectively balance their obligations to promote public safety with their traditional obligations to promote the best interests of their individual patients. To meet these concurrent ethical obligations, physicians' participation in public health violence prevention should seek to improve public safety without compromising the care of patients or exposing individuals to undue harm. Physicians should, therefore, report to the appropriate authorities those patients who are at risk of committing violent acts toward the public, but should only disclose the minimal amount of information that is necessary to protect the public. Moreover, physicians should also recommend the separation of violent individuals from the community at large when necessary to improve public safety while advocating for the provision of appropriate treatment measures to improve the patients' well being. PMID- 18388614 TI - Safer schools in an age of mass violence: back to the basics of public health. PMID- 18388615 TI - Protecting the public's health following the Virginia Tech tragedy: issues of law and policy. AB - Assessing legal responsibility in the aftermath of the April 2007 tragedy at Virginia Polytechnic Institute and State University (Virginia Tech) is inevitable. Beyond assigning blame, law- and policymakers should examine ways to protect the public from future incidences of gun violence on campuses and other settings. Although no combination of legal responses may fully deter individuals who are intent on causing significant harm, select legal reforms have the potential to prevent future acts of gun violence. These reforms include considering more restrictive gun laws nationally, reporting individuals with known mental impairments that may endanger themselves or others to federal or state databases, and refining laws that limit institutions from acting in advance to address prospectively dangerous people. Each of these reforms has the potential to reduce acts of gun violence to improve the public's health, but also implicates individual rights and interests. PMID- 18388616 TI - State of security at US colleges and universities: a national stakeholder assessment and recommendations. AB - In 2004 the US Department of Justice, Office of Community Oriented Policing Services, sponsored a National Summit on Campus Public Safety. The summit brought together various stakeholders including campus police and security officials, local police chiefs, college and university faculty and administrators, federal officials, students and parents, and community leaders to address the issues and complexities of campus safety. Delegates to the summit identified key issues in campus safety and security, which included establishing a national center on campus safety, balancing traditional open environments with the need to secure vulnerable sites, improving coordination with state and local police, reducing internal fragmentation, elevating professionalism, and increasing eligibility of campus police and security agencies to compete for federal law enforcement funds. Focus on "active shooters" on campus, resulting from the Virginia Tech incident, should not diminish attention placed on the broader, more prevalent safety and security issues facing the nation's educational campuses. Recommendations resulting from the summit called for establishing a national agenda on campus safety, formation of a national center on campus public safety, and increased opportunity for campus police and security agencies to compete for federal and state funds. PMID- 18388617 TI - Developing a consensus framework for an effective and efficient disaster response health system: a national call to action. AB - Eighteen national organizations, representing medicine, dentistry, nursing, hospital systems, public health, and emergency medical services, have worked together to create a framework for a national and regional disaster response health system that is scalable, multidisciplinary, and seamless, and based on an all-hazards approach. In July 2005 and June 2006 the American Medical Association (AMA) and the American Public Health Association (APHA) convened the AMA/APHA Linkages Leadership Summit, with funding from the Centers for Disease Control and Prevention under the Terrorism Injuries: Information Dissemination and Exchange (TIIDE) program. As cofacilitators, James J. James, MD, DrPH, MHA, director of the AMA Center for Public Health Preparedness and Disaster Response, and Georges Benjamin, MD, FACP, FACEP(E), APHA executive director, met with leaders from 16 national medical, dental, hospital, nursing, hospital systems, public health, and emergency medical services organizations in Chicago (2005) and New Orleans (2006) to deliberate the deficiencies in the medical and public health disaster response system and the lack of necessary linkages between key components of this system: the health care, emergency medical services, and public health sectors. The goal was to reach consensus on a set of overarching recommendations to improve and sustain health system preparedness and to combine each organization's advocacy expertise and experience to promote a shared policy agenda. The full summit report contains 53 consensus-based recommendations, which will serve as the framework for a coordinated national agenda for strengthening health system preparedness for terrorism and other disasters. The 9 most overarching critical recommendations from the report are highlighted here. Although the summit report presents important perspectives on the subject of preparedness for public health emergencies, we must understand that preparedness is a process and that these recommendations must be reviewed and refined continually over time. PMID- 18388618 TI - Role of tactical EMS in support of public safety and the public health response to a hostile mass casualty incident. PMID- 18388619 TI - Active shooters on college campuses: conflicting advice, roles of the individual and first responder, and the need to maintain perspective. PMID- 18388620 TI - A nation prepared: inspiration in the face of tragedy. PMID- 18388621 TI - Lessons from the response to the Virginia Tech shootings. PMID- 18388622 TI - Regional health system response to the Virginia Tech mass casualty incident. AB - BACKGROUND: On April 16, 2007 a mass shooting occurred on the campus of Virginia Polytechnic Institute and State University (Virginia Tech). Due to both distance and weather, air transport of the injured directly to a level 1 trauma center was not possible. The injured received all of their care or were initially stabilized at 3 primary hospitals that either had a level 3 trauma center designation or no trauma center designation. METHODS: This article is a retrospective analysis of the regional health system (prehospital, hospital, regional hospital emergency operations center, and public health local and state) response. Data records from all of the regional responding emergency medical services, hospitals, and coordinating services were reviewed and analyzed. Records for all 26 patients were reviewed and analyzed using triage designations, injury severity scores (ISS), and critical mortality. RESULTS: Twenty-five of the 26 patients were triaged in the field. Excluding 1 patient (asthma), the average ISS for victims presenting was 8.2. Twelve patients had an ISS of > or = 9, and 5 had an ISS score of > or = 15. Ten of the 26 patients (38%) required urgent intervention and surgery in the first 24 hours. The overall regional health system mortality of victims received was 3.8% (1 death [excluding 1 dead on arrival {DOA}]/ 26 victims from scene). The regional health system critical mortality rate (excluding 1 victim who was DOA) was 20% (1/5). DISCUSSION: The outcomes of the Virginia Tech mass casualty incident, as evidenced by the low overall regional health system mortality of victims received at 3.8% (1/26) and low critical mortality rate (excluding 1 victim who was DOA) of 20%, coupled with a need to treat a significant amount of moderately injured victims 46% (12/26 with ISS > or = 9) gives credence to the successful response. The successful response occurred as a consequence of regional collaborative planning, training, and exercising, which resulted not only in increased expertise and improved communications but also in essential relationships and a sense of trust forged among all of the responders. PMID- 18388624 TI - Re: Prediction modeling to determine the adequacy of medical response to urban nuclear attack. PMID- 18388625 TI - The greatest danger. PMID- 18388626 TI - Tornadoes and disaster management: not just in Kansas, Dorothy. PMID- 18388627 TI - Dilemmas of neglected humanitarian crises. PMID- 18388628 TI - Successful collaboration of medicine and public health. PMID- 18388629 TI - Mature and well-thought out plan. PMID- 18388630 TI - Increased collective ability to apply research findings to emergency preparedness and response. PMID- 18388631 TI - Practitioner liability protections approved. PMID- 18388632 TI - Pandemic influenza community preparedness planning: Second National Congress on Health System Readiness. PMID- 18388633 TI - World update. PMID- 18388634 TI - Prediction modeling to determine the adequacy of medical response to urban nuclear attack. AB - BACKGROUND: Government reports have persistently indicated the intent of terrorists and hostile nations to acquire and "weaponize" nuclear materials for deliberate attack on a major US metropolitan city. METHODS: A modeling analysis of the effects of 20- and 550-kiloton nuclear detonations on the 2 major metropolitan centers of Los Angeles and Houston is presented with a focus on thermal casualties. Brode's work as modified by Binninger was used to calculate thermal fluence, using thermal fractions. The EM-1 and WE programs were used to calculate blast effects. Fallout radiation was calculated using the Defense Threat Reduction Agency's Hazard Prediction and Assessment Capability V404SP4 with "urban effects" turned on. The ESRI ArcView program calculated affected populations from 2000 US Census block-level data for areas affected by thermal effects. RESULTS: The population affected by a 550-kiloton nuclear weapon detonated in Los Angeles and Houston is staggering: surviving thermal casualties are estimated at 185,000 and 59,000, respectively. Even the 20-kiloton detonations in Los Angeles and Houston are significant: the numbers of surviving thermal casualties requiring care exceed 28,000 and 10,000, respectively. CONCLUSIONS: The surviving health care community postdetonation would be faced with an unprecedented burden of care for thermal casualties. A great expansion of personnel involved in emergency burn care response is critical. Bold, new approaches such as regionalization and predetermined medical air transport need to be considered. PMID- 18388635 TI - Regional Health System Response to the 2007 Greensburg, Kansas, EF5 Tornado. AB - BACKGROUND: On May 4, 2007 an EF5 tornado hit the rural community of Greensburg, KS, destroying 95% of the town and resulting in 12 fatalities. METHODS: Data was requested from the emergency medical services units that initially responded and the regional hospitals that received people injured in the tornado within 24 hours following the tornado. Requested data included patient age and sex, and injury severity score or ICD-9 codes. Critical mortality, or the number of deaths of critically injured patients, was also calculated. RESULTS: The extensive damage caused by the tornado effectively destroyed the infrastructure of the community and created enormous challenges for emergency medical services responders, who were unable to record any triage data. Area hospitals treated 90 patients, who had an average injury severity score of 6.4. Age was found to be related to injury severity, but no relationship between sex and injury severity was found. Critical mortality was found to be 18% for this event. CONCLUSIONS: Injury severity score has seldom been used to analyze natural disasters, especially tornadoes, although such analysis is helpful for understanding the magnitude of the disaster, comparing to other disasters, and preparing for future incidents. Advanced warning and personal preparedness are important factors in reducing tornado-related injuries and deaths. PMID- 18388636 TI - Assessing the integration of health center and community emergency preparedness and response planning. AB - BACKGROUND: To assess the state of health center integration into community preparedness, we undertook a national study of linkages between health centers and the emergency preparedness and response planning initiatives in their communities. The key objectives of this project were to gain a better understanding of existing linkages in a nationally representative sample of health centers, and identify health center demographic and experience factors that were associated with strong linkages. METHODS: The objectives of the study were to gain a baseline understanding of existing health center linkages to community emergency preparedness and response systems and to identify factors that were associated with strong linkages. A 60-item questionnaire was mailed to the population of health centers supported by the Health Resources and Services Administration's Bureau of Primary Health Care in February 2005. Results were aggregated and a chi square analysis identified factors associated with stronger linkages. RESULTS: Overall performance on study-defined indicators of strong linkages was low: 34% had completed a hazard vulnerability analysis in collaboration with the community emergency management agency, 30% had their role documented in the community plan, and 24% participated in community-wide exercises. Stronger linkages were associated with experience responding to a disaster and a perception of high risk for experiencing a disaster. CONCLUSIONS: The potential for health centers to participate in an integrated response is not fully realized, and their absence from community-based planning leaves an already vulnerable population at greater risk. Community planners should be encouraged to include health centers in planning and response and centers should receive more targeted resources for community integration. PMID- 18388637 TI - Postexposure immunization and prophylaxis of bloodborne pathogens following a traumatic explosive event: preliminary recommendations. AB - BACKGROUND: No definitive guidelines have been established in the United States for postexposure immunization and prophylaxis (PEP) to hepatitis B and C viruses (HBV, HCV) and human immunodeficiency virus (HIV) in the event of a traumatic explosive event. METHODS: The American Medical Association's Center for Public Health Preparedness and Disaster Response assembled a US-Israeli panel of experts, including representatives from disaster medicine, trauma surgery, occupational health, and infectious disease to determine guidelines for adult and pediatric victims following a traumatic explosive event. The panel reviewed the existing Israeli and United Kingdom protocols, previously published Centers for Disease Control and Prevention guidance on occupational and nonoccupational exposures to HBV, HCV, and HIV, before reaching consensus on preliminary guidelines for the United States. RESULTS: These guidelines recommend an age appropriate dose and schedule for HBV PEP for individuals presenting from the scene with nonintact skin or mucous membrane exposure, and they also consider HCV and HIV testing in individuals presenting with possible nonintact skin or mucous membrane exposure. The guidelines do not recommend PEP for individuals presenting from the scene with possible superficial skin exposure. CONCLUSIONS: These recommendations offer PEP guidance for bloodborne pathogens and are limited in scope. These recommendations do not address general wound PEP such as tetanus or the need for antibiotics. It is hoped that these guidelines will fill an urgent gap in preparedness until definitive, comprehensive guidelines from the Centers for Disease Control and Prevention are published. PMID- 18388638 TI - Measuring humanitarian emergencies. AB - The ability to monitor assistance, define humanitarian needs, and approach equity in the distribution of assistance has lagged behind the world's growing commitment to responding to humanitarian emergencies. This article highlights relevant data sources to elucidate elements of an operational definition of humanitarian need. New and refined measures are proposed to assist in assessing the level of need among affected populations. An original measure that combines data on conflict and disasters to summarize the cumulative magnitude of 4 types of humanitarian threats is presented. PMID- 18388639 TI - The new International Health Regulations: considerations for global public health surveillance. AB - Global public health surveillance is critical for the identification and prevention of emerging and reemerging infectious diseases. The World Health Organization recently released revised International Health Regulations (IHR) that serve as global legislation and provide guidelines for surveillance systems. The IHR aim to identify and prevent spread of these infectious diseases; however, there are some practical challenges that limit the usability of these regulations. IHR requires Member States to build necessary infrastructure for global surveillance, which may not be possible in underdeveloped countries. A large degree of freedom is given to each individual government and therefore different levels of reporting are common, with substantial emphasis on passive reporting. The IHR need to be enforceable and enforced without impinging on government autonomy or human rights. Unstable governments and developing countries require increased assistance in setting up and maintaining surveillance systems. This article addresses some challenges and potential solutions to the ability of national governments to adhere to the global health surveillance requirements detailed in the IHR. The authors review some practical challenges such as inadequate surveillance and reporting infrastructure, and legal enforcement and maintenance of individual human rights. PMID- 18388640 TI - Botulism: cause, effects, diagnosis, clinical and laboratory identification, and treatment modalities. AB - Botulism is a neuroparalytic disease caused by neurotoxins produced by the bacteria Clostridium botulinum. Botulinum neurotoxins (BoNTs) are among the most potent naturally occurring toxins and are a category A biological threat agent. The 7 toxin serotypes of BoNTs (serotypes A-G) have different toxicities, act through 3 different intracellular protein targets, and exhibit different durations of effect. Botulism may follow ingestion of food contaminated with BoNT, from toxin production of C botulinum present in the intestine or wounds, or from inhalation of aerosolized toxin. Intoxication classically presents as an acute, symmetrical, descending flaccid paralysis. Early diagnosis is important because antitoxin therapy is most effective when administered early. Confirmatory testing of botulism with BoNT assays or C botulinum cultures is time-consuming, and may be insensitive in the diagnosis of inhalational botulism and in as many as 32% of food-borne botulism cases. Therefore, the decision to initiate botulinum antitoxin therapy is primarily based on symptoms and physical examination findings that are consistent with botulism, with support of epidemiological history and electrophysiological testing. Modern clinical practice and antitoxin treatment has reduced botulism mortality rates from approximately 60% to < or =10%. The pentavalent botulinum toxoid is an investigational product and has been used for more than 45 years in at-risk laboratory workers to protect against toxin serotypes A to E. Due to declining immunogenicity and potency of the pentavalent botulinum toxoid, novel vaccine candidates are being developed. PMID- 18388641 TI - Definition and functions of health unified command and emergency operations centers for large-scale bioevent disasters within the existing ICS. AB - The incident command system provides an organizational structure at the agency, discipline, or jurisdiction level for effectively coordinating response and recovery efforts during most conventional disasters. This structure does not have the capacity or capability to manage the complexities of a large-scale health related disaster, especially a pandemic, in which unprecedented decisions at every level (eg, surveillance, triage protocols, surge capacity, isolation, quarantine, health care staffing, deployment) are necessary to investigate, control, and prevent transmission of disease. Emerging concepts supporting a unified decision-making, coordination, and resource management system through a health-specific emergency operations center are addressed and the potential structure, function, roles, and responsibilities are described, including comparisons across countries with similar incident command systems. PMID- 18388642 TI - Linking public health and the emergency care community: 7 model communities. AB - Public health and the emergency care community must work together to effectively achieve a state of community-wide disaster preparedness. The identification of model communities with good working relationships between their emergency care community and public health agencies may provide useful information on establishing and strengthening relationships in other communities. Seven model communities were identified: Boston, Massachusetts; Clark County, Nevada; Eau Claire, Wisconsin; Erie County, New York; Louisville, Kentucky; Livingston County, New York; and Monroe County, New York. This article describes these communities and provides a summary of common findings. Specifically, we recommend that communities foster respectful working relationships between agency leaders, hold regular face-to-face meetings, educate each other on their expertise and roles during a disaster, develop response plans together, work together on a day to-day basis, identify and encourage a leader to facilitate these relationships, and share resources. PMID- 18388646 TI - Re: Letter from a physician whose practice was devastated by Hurricane Katrina. PMID- 18388647 TI - Is overtriage associated with increased mortality? The evidence says "yes". PMID- 18388649 TI - Allocating mechanical ventilators during mass respiratory failure: Kudos to New York State, but more work to be done. PMID- 18388650 TI - Disaster medicine: what's the reality? PMID- 18388651 TI - New presidential directive targets academic approaches to disasters. PMID- 18388652 TI - 2007 San Diego County wildfire response: a lesson in disaster preparedness collaboration. PMID- 18388653 TI - World update. PMID- 18388654 TI - Allocation of ventilators in a public health disaster. AB - BACKGROUND: In a public health emergency, many more patients could require mechanical ventilators than can be accommodated. METHODS: To plan for such a crisis, the New York State Department of Health and the New York State Task Force on Life and the Law convened a workgroup to develop ethical and clinical guidelines for ventilator triage. RESULTS: The workgroup crafted an ethical framework including the following components: duty to care, duty to steward resources, duty to plan, distributive justice, and transparency. Incorporating the ethical framework, the clinical guidelines propose both withholding and withdrawing ventilators from patients with the highest probability of mortality to benefit patients with the highest likelihood of survival. Triage scores derive from the sepsis-related organ failure assessment system, which assigns points based on function in 6 basic medical domains. Triage may not be implemented by a facility without clear permission from public health authorities. CONCLUSIONS: New York State released the draft guidelines for public comment, allowing for revision to reflect both community values and medical innovation. This ventilator triage system represents a radical shift from ordinary standards of care, and may serve as a model for allocating other scarce resources in disasters. PMID- 18388655 TI - Chronic disease and related conditions at emergency treatment facilities in the New Orleans area after Hurricane Katrina. AB - BACKGROUND: Disaster preparations usually focus on preventing injury and infectious disease. However, people with chronic disease and related conditions (CDRCs), including obstetric/gynecological conditions, may be vulnerable to disruptions caused by disasters. METHODS: We used surveillance data collected after Hurricane Katrina to characterize the burden of visits for CDRCs at emergency treatment facilities (eg, hospitals, disaster medical assistance teams, military aid stations). In 6 parishes in and around New Orleans, health care providers at 29 emergency treatment facilities completed a standardized questionnaire for injury and illness surveillance from September 8 through October 22, 2005. RESULTS: Of 21,673 health care visits, 58.0% were for illness (24.3% CDRCs, 75.7% non-CDRCs), 29.1% for injury, 7.2% for medication refills, and 5.7% for routine or follow-up care. The proportion of visits for CDRCs increased with age. Among men presenting with CDRCs, the most common illnesses were cardiovascular disease (36.8%), chronic lower-respiratory disease (12.3%), and diabetes/glucose abnormalities (7.7%). Among women presenting with CDRCs, the most common were cardiovascular disease (29.2%), obstetric/gynecological conditions (18.2%), and chronic lower-respiratory disease (12.0%). Subsequent hospitalization occurred among 28.7% of people presenting with CDRCs versus 10.9% of those with non-CDRCs and 3.8% of those with injury. CONCLUSIONS: Our data illustrate the importance of including CDRCs as a part of emergency response planning. PMID- 18388656 TI - Trial of prophylactic inhaled steroids to prevent or reduce pulmonary function decline, pulmonary symptoms, and airway hyperreactivity in firefighters at the world trade center site. AB - BACKGROUND: Inhaled corticosteroids (ICS) are the most effective anti inflammatory treatment for asthmatics. This trial evaluated the effects of prophylactic ICS in firefighters exposed to the World Trade Center disaster. METHODS: Inhaled budesonide via a dry powder inhaler (Pulmicort Turbuhaler, AstraZeneca, Wilmington, DE) was offered on-site to New York City firefighters between September 18 and 25, 2001. One to 2 years later, firefighters (n = 64) who completed 4 weeks of daily ICS treatment were evaluated and compared with an age- and exposure-matched comparison group (n = 72) who did not use ICS. RESULTS: When spirometry results at the final visit were compared with those from the weeks following the 9/11 disaster, the treatment group had a greater increase in forced vital capacity (P = .009) and possibly a slower decline in forced expiratory volume at 1 second (P = .11), as well as a greater improvement in perceived well-being as assessed by the St George's Respiratory Questionnaire (P < .01). There was no difference in airway hyperreactivity and no evidence of adverse effects from ICS. CONCLUSIONS: Because the potential for hazardous exposures is great at many disasters, disease prevention programs based on environmental controls and respiratory protection are warranted immediately. Our results suggest that, pending further study with a larger sample, prophylactic ICS should be considered, along with respiratory protection, to minimize possible lung insult. PMID- 18388657 TI - Implementing the cities readiness initiative: lessons learned from Boston. AB - The federally funded Cities Readiness Initiative (CRI) requires seamless federal, state, and local public health coordination to provide antibiotics to an entire city population within 48 hours of an aerosolized release of anthrax. We document practical lessons learned from the development and implementation of the Boston CRI plan. Key themes center on heightened emphasis on security, a new mass protection model of dispensing, neighborhood-centric clinic site selection, online training of Medical Reserve Corps volunteers, and the testing of operations through drills and exercises. Sharing such lessons can build national preparedness. PMID- 18388658 TI - Addressing gaps in health care sector legal preparedness for public health emergencies. AB - Health care providers and their legal counsel play pivotal roles in preparing for and responding to public health emergencies. Lawyers representing hospitals, health systems, and other health care provider components are being called upon to answer complex legal questions regarding public health preparedness issues that most providers have not previously faced. Many of these issues are legal issues with which public health officials should be familiar, and that can serve as a starting point for cross-sector legal preparedness planning involving both the public health and health care communities. This article examines legal issues that health care providers face in preparing for public health emergencies, and steps that providers, their legal counsel, and others can take to address those issues and to strengthen community preparedness. PMID- 18388659 TI - A consensus-based educational framework and competency set for the discipline of disaster medicine and public health preparedness. AB - BACKGROUND: Various organizations and universities have developed competencies for health professionals and other emergency responders. Little effort has been devoted to the integration of these competencies across health specialties and professions. The American Medical Association Center for Public Health Preparedness and Disaster Response convened an expert working group (EWG) to review extant competencies and achieve consensus on an educational framework and competency set from which educators could devise learning objectives and curricula tailored to fit the needs of all health professionals in a disaster. METHODS: The EWG conducted a systematic review of peer-reviewed and non-peer reviewed published literature. In addition, after-action reports from Hurricane Katrina and relevant publications recommended by EWG members and other subject matter experts were reviewed for congruencies and gaps. Consensus was ensured through a 3-stage Delphi process. RESULTS: The EWG process developed a new educational framework for disaster medicine and public health preparedness based on consensus identification of 7 core learning domains, 19 core competencies, and 73 specific competencies targeted at 3 broad health personnel categories. CONCLUSIONS: The competencies can be applied to a wide range of health professionals who are expected to perform at different levels (informed worker/student, practitioner, leader) according to experience, professional role, level of education, or job function. Although these competencies strongly reflect lessons learned following the health system response to Hurricane Katrina, it must be understood that preparedness is a process, and that these competencies must be reviewed continually and refined over time. PMID- 18388661 TI - Viagra and wound healing: the NO connection. PMID- 18388665 TI - 2008 wound-related coding changes. PMID- 18388666 TI - Does the proximity of an amputation, length of time between foot ulcer development and amputation, or glycemic control at the time of amputation affect the mortality rate of people with diabetes who undergo an amputation? AB - OBJECTIVES: The main purpose of the authors' research was to compare the 3- and 5 year mortality rates of diabetic patients who have undergone a lower-extremity amputation, evaluating the proximity of the amputation in 3 groups (toe and foot amputation, BKA, and AKA), the timing of the amputation (within or after 2 years of the onset of the foot ulcer), and the effect of glycemic control at the time of amputation, regardless of the level of amputation. METHODS AND DESIGN: The subjects were 80 male inpatients at Illiana Veterans Health Care System who had diabetes, diabetic neuropathy, peripheral vascular disease, and a diabetic foot ulcer that resulted in an amputation. Of the 80 patients, 29 had a toe amputation, 30 had a BKA, and 21 had an AKA. The mean age in all 3 groups was 68.5 years+/-7.2 years. The authors used the hemoglobin A1C (Hgb A1C) level to assess glycemic control. MAIN OUTCOME MEASURES: Several chi tests were used to compare the 3- and 5-year mortality rates among the amputation groups. An Hgb A1C level of 8% or less defined good glycemic control; an Hgb A1C level of more than 8% defined poor glycemic control. A chi test was used to compare glycemic control, the level of amputation, and the mortality rate. A chi test was also used to evaluate the length of time between ulcer formation and amputation, the level of amputation, and the mortality. All statistics were done using SPSS 10.0 student version. CONCLUSIONS: The research revealed a statistically significant difference in mortality among the 3 groups at 3 and 5 years. No statistically significant difference in mortality in patients with good glycemic control and patients with bad glycemic control was found. At 3 years, a statistically significant difference in mortality existed between patients who had an amputation within 2 years of ulcer formation and those who had an amputation after 2 years. At 5 years, no statistically significant difference existed between these 2 groups. PMID- 18388667 TI - Multisite Web-based training in using the Braden Scale to predict pressure sore risk. AB - OBJECTIVE: To evaluate the effect of a Web-based Braden Scale training module on nurses' knowledge of pressure-ulcer risk assessment and prevention. DESIGN: Pre experimental, posttest-only design. SETTING: Web-based learning environment. PARTICIPANTS: Registered nurses (N=1391) working at 3 medical centers in the Midwest. MAIN OUTCOME MEASURES: Primary outcomes of interest were reliability and competence associated with using the Braden Scale for pressure-ulcer risk assessment. Secondary outcomes of interest focused on program evaluation, specifically nurses' perceptions of program adequacy and ease of use. MAIN RESULTS: After training, nurses correctly rated Braden Scale level of risk 82.6% of the time. Numeric ratings for Braden subscales were generally more reliable when case-study data indicated extreme risk levels (generally not at-risk level, high-risk level, and very high level) than when data indicated midlevels of risk (mild-risk level and moderate-risk level). Nurses' knowledge of appropriate risk based preventive interventions was high, but correlated poorly with the ability to correctly assign numeric ratings to Braden subscales. CONCLUSION: Web-based training alone may not ensure reliable, competent estimates of pressure-ulcer risk for patients at all risk levels. Other strategies, such as clinical practice with expert supervision, should be considered. Further research is needed to clarify the links between scoring Braden subscales correctly and selecting appropriate risk-based preventive interventions. PMID- 18388668 TI - A new era of pressure ulcer accountability in acute care. AB - PURPOSE: To familiarize wound care practitioners with the 5 Million Lives Campaign's pressure ulcer prevention intervention. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in wound care. OBJECTIVES: After reading this article and taking this test, the reader should be able to: 1. List organizational and national initiatives that address pressure ulcer prevention. 2. Identify characteristics of pressure ulcers. 3. Identify 6 strategies that help prevent pressure ulcers. PMID- 18388670 TI - Managing tissue loads. PMID- 18388672 TI - Neuroscience. PMID- 18388673 TI - Treatment of intracranial hypertension. AB - PURPOSE OF REVIEW: The review provides key points and recent advances regarding the treatments of intracranial hypertension as a consequence of traumatic brain injury. The review is based on the pathophysiology of brain edema and draws on the current literature as well as clinical bedside experience. RECENT FINDINGS: The review will cite baseline literature and discuss emerging data on cerebral perfusion pressure, sedation, hypothermia, osmotherapy and albumin as treatments of intracranial hypertension in traumatic brain-injured patients. SUMMARY: One of the key issues is to consider that traumatic brain injury is more likely a syndrome than a disease. In particular, the presence or absence of a high contusional volume could influence the treatments to be implemented. The use of osmotherapy and/or high cerebral perfusion pressure should be restricted to patients without major contusions. Some physiopathological, experimental and clinical data, however, show that corticosteroids and albumin--therapies that have been proven deleterious if administered systematically--are worth reconsidering for this subgroup of patients. The current Pitie-Salpetriere algorithm, where treatments are stratified according to their potential side effects, will be added at the end of the review as an example of an integrated strategy. PMID- 18388674 TI - Biomarkers of primary and evolving damage in traumatic and ischemic brain injury: diagnosis, prognosis, probing mechanisms, and therapeutic decision making. AB - PURPOSE OF REVIEW: Emerging data suggest that biomarkers of brain injury have potential utility as diagnostic, prognostic, and therapeutic adjuncts in the setting of traumatic and ischemic brain injury. Two approaches are being used, namely, assessing markers of structural damage and quantifying mediators of the cellular, biochemical, or molecular cascades in secondary injury or repair. Novel proteomic, multiplex, and lipidomic methods are also being applied. RECENT FINDINGS: Biochemical markers of neuronal, glial, and axonal damage such as neuron-specific enolase, S100B, and myelin basic protein, respectively, are readily detectable in biological samples such as serum or cerebrospinal fluid and are being studied in patients with ischemic and traumatic brain injury. In addition, a number of studies have demonstrated that novel tools to assess simultaneously multiple biomarkers can provide unique insight such as details on specific molecular participants in cell death cascades, inflammation, or oxidative stress. SUMMARY: Multifaceted cellular, biochemical, and molecular monitoring of proteins and lipids is logical as an adjunct to guiding therapies and improving outcomes in traumatic and ischemic brain injury and we appear to be on the verge of a breakthrough with the use of these markers as diagnostic, prognostic, and monitoring adjuncts, in neurointensive care. PMID- 18388675 TI - Prehospital intubation of brain-injured patients. AB - PURPOSE OF REVIEW: The article reviews the evidence for and against early intubation for brain-injured patients. RECENT FINDINGS: Although theoretical advantages to early intubation include airway protection, improved oxygenation, and control of ventilation, recent clinical data document an association between early intubation and mortality. This likely reflects some degree of selection bias when considering intubation without the use of neuromuscular blocking agents. Paramedic use of these drugs, however, has also been associated with higher mortality. Subgroup analysis from the San Diego Paramedic Rapid Sequence Intubation Trial and outcome data following air medical intubation suggest that suboptimal performance of intubation and subsequent ventilation may play an important role in determining the procedure's potential benefit or harm. In addition, refining patient selection criteria for early intubation may result in better outcomes. SUMMARY: Available evidence is inconclusive regarding the role for early intubation following acute brain injury. Suboptimal performance of intubation and subsequent ventilation may offset potential benefits of the procedure. In addition, studies to better define the patient population benefiting from early intubation will help avoid exposing other patients to a potentially dangerous procedure. PMID- 18388676 TI - Venous thromboembolism prophylaxis and treatment in patients with acute stroke and traumatic brain injury. AB - PURPOSE OF REVIEW: Patients with acute stroke and traumatic brain injury are at risk to develop venous thromboembolism. This review analyzes the available literature to propose guidelines for the prevention and treatment of venous thromboembolism in these groups of patients. RECENT FINDINGS: In acute ischemic stroke, low-dose low-molecular-weight heparin has the best benefit-risk ratio to prevent venous thromboembolism. Patients with primary intracerebral hemorrhage and traumatic brain injury should receive intermittent pneumatic compression, followed by low-dose low-molecular-weight heparin or unfractioned heparin 3-4 days after stroke onset or 24 h after injury or surgery, respectively, and after cessation of bleeding. Concerning treatment, in patients with deep-vein thrombosis lower doses of heparin are indicated to prevent pulmonary embolism, and a vena cava filter should be considered. In patients with pulmonary embolism, treatment could be more aggressive, because of a high mortality risk. SUMMARY: Adequate prevention of venous thromboembolism with intermittent pneumatic compression or pharmacological prophylaxis is important. The best treatment of venous thromboembolism remains unclear. In case of pulmonary embolism, more aggressive treatment is warranted. PMID- 18388677 TI - Optimal hemoglobin concentration in patients with subarachnoid hemorrhage, acute ischemic stroke and traumatic brain injury. AB - PURPOSE OF REVIEW: The review outlines recent clinical and experimental studies regarding the effects of red blood-cell transfusion on clinical outcome in neurocritical patients, including patients with subarachnoid hemorrhage, acute ischemic stroke and traumatic brain injury. Optimal hemoglobin transfusion trigger and the role of other transfusion indicators for neurocritical patients are discussed. RECENT FINDINGS: Acute anemia (hemoglobin levels near 7 g/dl) is well tolerated by healthy subjects, but extreme anemia might negatively affect clinical outcome of neurocritical patients. Conversely, high hemoglobin levels, attained by means other than red blood-cell transfusion, improve clinical outcome, whereas red blood-cell transfusion is associated with poorer clinical outcome (mortality, length of stay and disability) in patients presenting subarachnoid hemorrhage, acute ischemic stroke and traumatic brain injury. Studies defining the optimal hemoglobin concentration in neurocritical patients are lacking, but a restrictive transfusion policy seems to be safe and is often recommended. In the near future, signals coming from the brain, such as brain tissue oxygen tension and regional cerebral oxygen saturation, might potentially be developed into transfusion triggers. SUMMARY: Both severe anemia and red blood cell transfusion may negatively influence clinical outcome in neurocritical patients. Acceptance of low hemoglobin concentrations may be justified by avoiding negative transfusion effects. No evidence-based transfusion trigger in neurocritical patients can be recommended. PMID- 18388678 TI - Adrenal insufficiency following traumatic brain injury in adults. AB - PURPOSE OF REVIEW: Hypoadrenalism occurs in approximately 25% of patients soon after traumatic brain injury. Neurosurgeons or critical care physicians should be prepared to diagnose and treat this and other related hormonal deficiencies. RECENT FINDINGS: The severity of traumatic brain injury, location of basilar skull fractures and edema or hemorrhage within the hypothalamic-pituitary axis appear correlated with secondary adrenal failure. Primary hypoadrenalism also may occur due to injury-related systemic inflammation. Hypotension requiring vasoactive drug support, hyponatremia and hypoglycemia may be corresponding clinical signs. Evaluation of either primary or secondary hypoadrenalism should include measurement of basal and post-adrenocorticotropin stimulation cortisol blood concentrations. If the basal cortisol is under 15 microg/dl or increases by over 9 microg/dl after stimulation treatment should be considered. Intravenous hydrocortisone at 50-100 mg every 8 h or by continuous infusion is usually sufficient but may be supplemented with a mineralocorticoid if hyponatremia persists. All patients sustaining severe traumatic brain injury should be tested for endocrine failure (adrenal, thyroid and growth hormone) 3 months after injury. SUMMARY: Adrenal gland failure or the inability to produce adrenocorticotropin and other pituitary hormones may occur early after traumatic brain injury. Acute treatment of either cause of hypoadrenalism may correct associated hypotension, hypoglycemia, or hyponatremia. PMID- 18388679 TI - Hyperoxia: good or bad for the injured brain? AB - PURPOSE OF REVIEW: For decades it was assumed that cerebral ischemia was a major cause of secondary brain injury in traumatic brain injury, and management focused on improving cerebral perfusion and blood flow. Following the observation of mitochondrial dysfunction in traumatic brain injury and the widespread use of brain tissue oxygen tension (P(br)O(2) monitoring, however, recent work has focused on the use of hyperoxia to reduce the impact of traumatic brain injury. RECENT FINDINGS: Previous work on normobaric hyperoxia utilized very indirect measures of cerebral oxygen metabolism (intracranial pressure, brain oxygen tension and microdialysis) as outcome variables. Interpretation of these measures is controversial, making it difficult to determine the impact of hyperoxia. A recent study, however, utilized positron emission tomography to study the impact of hyperoxia on patients with acute severe traumatic brain injury and found no improvement on cerebral metabolic rate for oxygen with this intervention. SUMMARY: Despite suggestive data from microdialysis studies, direct measurement of the ability of the brain to utilize oxygen indicates that hyperoxia does not increase oxygen utilization. This, combined with the real risk of oxygen toxicity, suggests that routine clinical use is not appropriate at this time and should await appropriate prospective outcome studies. PMID- 18388680 TI - Acute pancreatitis. AB - PURPOSE OF REVIEW: To review advances over the last year in the prevention, diagnosis and management of acute pancreatitis. RECENT FINDINGS: Obesity is an independent risk factor for severity in acute pancreatitis, and heavy alcohol consumption for the development of necrosis in severe acute pancreatitis. Biochemical markers have been further tested, including carbohydrate-deficient transferrin for the diagnosis of alcohol-induced acute pancreatitis, urinary trypsinogen-2 as a diagnostic marker for acute pancreatitis, and interleukin-6 and procalcitonin as markers of disease severity. A new, simple stratification system, the 'panc 3 score', has been described. There are conflicting data on the use of antibiotic prophylaxis in acute necrotizing pancreatitis, and on the chemoprevention of postendoscopic retrograde cholangiopancreatography pancreatitis. Enteral feeding is established as standard practice early in the management of acute pancreatitis of all aetiologies; probiotics and other compounds may also play a role. SUMMARY: Over the last year, there have been further innovations in the risk stratification and management of acute pancreatitis. Unresolved issues include chemoprevention of endoscopic retrograde cholangiopancreatography-induced acute pancreatitis, the indications for antibiotic prophylaxis in severe acute pancreatitis and nutritional supplementation with probiotics and synbiotics. PMID- 18388681 TI - Intensive care management of acute liver failure. AB - PURPOSE OF REVIEW: The mortality of acute liver failure remains unacceptably high and liver transplantation is the only effective treatment available to date. This review focuses on new research developments in the field and aims to provide a pragmatic organ-based treatment approach for liver failure patients requiring intensive care support. RECENT FINDINGS: The pathophysiological basis for cerebral edema formation in acute liver failure continued to be the focus of various investigations. In-vivo observations confirmed the link between ammonia, cerebral glutamine content and intracranial hypertension. The role of arterial ammonia as an important prognostic indicator formed the basis of prospective, observational studies. Reduced monocytic HLA-DR expression linked acute liver failure with poor prognosis, and the cerebral effects and side effects of vasoactive therapy with terlipressin were investigated with two studies showing contradictory results. SUMMARY: Despite increased knowledge of the pathophysiological events leading to organ dysfunction in acute liver failure, supportive treatment options remain limited in their efficacy and largely noncurative. PMID- 18388682 TI - Management of sepsis in patients with liver failure. AB - PURPOSE OF REVIEW: Sepsis constitutes the most common cause of death in the ICU. Liver dysfunction is manifested among previously normal subjects with sepsis but even more so in populations with preexisting liver disease. Managing these patients is more challenging. We will review recent literature in sepsis and liver disease, and their bedside application. RECENT FINDINGS: At the cellular chemical level, studies showed that platelet aggregation and neutrophil activation occur before and are independent of microcirculatory changes which are apparent in all animal septic models. At the clinical level, early goal-directed therapy, euglycemia, low tidal volume ventilation, and early and appropriately dosed renal replacement therapy among others are all tools to improve sepsis survival. Acknowledgement of liver disease as an immunocompromised host, and identification and treatment of complications can positively change the outcome of sepsis in liver disease. SUMMARY: Much has been advanced in the field of sepsis management. Understanding the pathophysiology of liver dysfunction and decompensation of a diseased liver incites questions for future research. Early goal-directed therapy, lactate clearance, glycemic control, low volume ventilation strategies, nutrition, adrenal insufficiency, renal dysfunction, hepatorenal syndrome prevention and treatment are some of the issues in the management of sepsis, with or without liver disease, that are relevant in this review. PMID- 18388683 TI - Etiologies of acute liver failure. AB - PURPOSE OF REVIEW: Acute liver failure remains a unique syndrome that results from massive loss of hepatocytes or hepatocyte function due to a variety of etiologies: viruses, drugs, toxins and other genetic and autoimmune conditions. The resulting clinical picture is remarkably similar across the different etiologies, reflecting common patterns of response of the innate immune system and the resulting inflammatory response. This review highlights developments in the delineation of etiologies of this relatively rare condition. RECENT FINDINGS: The contribution of different etiologies to the overall picture varies by country. Europe and North America demonstrate a large proportion of cases due to acetaminophen and to idiosyncratic drug reactions, whereas reports from emerging countries in Asia and Africa are more likely to feature viral illnesses, particularly hepatitis B and E. Recent studies have suggested ever more exotic etiologies, new drugs and viruses. SUMMARY: A careful search for the cause of acute liver failure in each case is of value in determining whether there are specific antidotes available and what the prognosis might be. PMID- 18388684 TI - Outcome of patients with cirrhosis admitted to intensive care. AB - PURPOSE OF REVIEW: The incidence of cirrhosis is increasing exponentially and is associated with significant morbidity and mortality. This cirrhotic population is prone to infection, which is a frequent precipitant for the development of organ dysfunction; a syndrome often referred to as 'acute-on-chronic' liver failure. Historically, the perception of cirrhosis with organ dysfunction as having a poor prognosis has led to invariably iniquitous access to intensive care. Data to support this view, however, are lacking. RECENT FINDINGS: Acute variceal bleeding is associated with markedly improved survival and warrants organ support in intensive care. Survival correlates directly with the number of organs failing, with sepsis and multiorgan failure resulting in over 90% mortality. The requirement for renal replacement therapy confers a poor prognosis in patients not suitable for liver transplantation. SUMMARY: Admission to intensive care for many patients with cirrhosis is not futile, particularly for those with single organ dysfunction and acute variceal bleeding. It can be extremely challenging to manage patients with organ dysfunction and encephalopathy in a ward environment, and these patients frequently require, and indeed benefit from, augmented levels of care in high-dependency and intensive care environments. PMID- 18388685 TI - Liver support devices. AB - PURPOSE OF REVIEW: Liver support devices are used either as a bridge to liver transplantation or liver recovery in patients with acute or acute-on-chronic liver failure. The review analyzes the recent literature and asks if the current enthusiasm for these devices is justified. RECENT FINDINGS: Many liver support devices exist and are discussed. Clinical data on artificial devices are rapidly emerging, especially on the molecular adsorbents recirculating system, and fractionated plasma separation and adsorption (Prometheus). While hepatic encephalopathy is improved by the molecular adsorbents recirculating system and probably Prometheus too, neither system has been shown to improve survival. Less clinical data exist for bioartificial support devices. These may use human hepatocytes, such as the extracorporeal liver assist device, although most devices use porcine hepatocytes, such as HepatAssist. SUMMARY: Enthusiasm in liver support devices is justified as many nonrandomized studies have suggested some biochemical and clinical benefits. The results of several ongoing multicenter randomized controlled trials are anxiously awaited. Meanwhile, because mortality without liver transplantation remains high despite the use of liver support devices, these devices should only be used in the research setting or by experts proficient in their use and as a bridge to liver transplantation rather than liver recovery. PMID- 18388686 TI - Bibliography. Current world literature. Neuroscience. PMID- 18388689 TI - Single gene contributions: genetic variants of peroxisome proliferator-activated receptor (isoforms alpha, beta/delta and gamma) and mechanisms of dyslipidemias. AB - PURPOSE OF REVIEW: Polymorphisms in peroxisome proliferator-activated receptor isoforms may be among the most important single-gene contributors to dyslipidemias, insulin resistance, and maturity-onset diabetes. RECENT FINDINGS: Familial partial lipodystrophy is a rare but characteristic phenotype associated with carriers of peroxisome proliferator-activated receptor-gamma missense mutations. Mutant receptors are transcriptionally defective, exhibit aberrant affinity for co-regulator molecules, and can exert dominant-negative or haplo insufficiency effects on normal peroxisome proliferator-activated receptor-gamma function. The P12A variant of isoform gamma is estimated to reduce diabetes risk by 19% in many populations, and has a large attributable risk because of high prevalence of the normal allele. Variants L162V and V227A of isoform alpha (common in white and Oriental populations, respectively) are associated with sexually dimorphic perturbations of lipid metabolism and cardiovascular risk. Polymorphisms in isoforms alpha and beta/delta are reported to influence lipid and glucose utilization. Apart from lipodystrophic syndromes, metabolic and cardiovascular risk in peroxisome proliferator-activated receptor variants is apparently modulated by dietary and exercise interventions, and interactions with polymorphisms in other genetic loci. SUMMARY: Polymorphisms in peroxisome proliferator-activated receptors are critical susceptibility risk factors for dyslipidemias and diabetes. They provide attractive targets for gene-environment interventions to reduce the burden of metabolic disease. PMID- 18388690 TI - Progress in the genetics of common obesity: size matters. AB - PURPOSE OF REVIEW: Over the past two decades serious efforts has been invested in the search for genes that predispose to common obesity, but progress has been slow and success limited. Genome-wide association, however, has revived optimism. Here we review recent advances in the field of obesity genetics and discuss the most important findings of candidate gene, genome-wide linkage studies and genome wide association studies. We conclude by speculating about the way forward in the near future. RECENT FINDINGS: Although large-scale candidate gene studies have placed MC4R more firmly on the human obesity map, the major breakthrough in obesity genetics was the discovery of FTO through genome-wide association. Variants located in the first intron of FTO were unequivocally associated with a 1.67-fold increased risk for obesity and a 0.40-0.66 kg/m2 increase in body mass index. SUMMARY: Genome-wide association promises to enhance greatly our understanding of the genetic basis of common obesity, although candidate gene studies will remain a valuable approach because they allow more detailed analyses of biologically relevant candidates. A key factor contributing to continued success lies in large-scale data integration through international collaboration, which will provide the sample sizes required to identify genetic association with conclusive evidence. PMID- 18388691 TI - Defining the spectrum of alleles that contribute to blood lipid concentrations in humans. AB - PURPOSE OF REVIEW: Recently, genome-wide genetic screening of common DNA sequence variants has proven a successful approach to identify novel genetic contributors to complex traits. This review summarizes recent genome-wide association studies for lipid phenotypes, and evaluates the next steps needed to obtain a full picture of genotype-phenotype correlation and apply these findings to inform clinical practice. RECENT FINDINGS: So far, genome-wide association studies have defined at least 19 genomic regions that contain common DNA single nucleotide polymorphisms associated with LDL cholesterol, HDL cholesterol and/or triglycerides. Of these, eight represent novel loci in humans, whereas 11 genes have been previously implicated in lipoprotein metabolism. Many of the same loci with common variants have already been shown to lead to monogenic lipid disorders in humans and/or mice, suggesting that a spectrum of common and rare alleles at each validated locus contributes to blood lipid concentrations. SUMMARY: At least 19 loci harbor common variations that contribute to blood lipid concentrations in humans. Larger scale genome-wide association studies should identify additional loci, and sequencing of these loci should pinpoint all relevant alleles. With a full catalog of DNA polymorphisms in hand, a panel of lipid-related variants can be studied to provide clinical risk stratification and targeting of therapeutic interventions. PMID- 18388692 TI - Cardiovascular disease risk prediction using genetic information (gene scores): is it really informative? AB - PURPOSE OF REVIEW: DNA-based tests for assessment of genetic predisposition to coronary heart disease need to provide information over and above that of conventional risk factors. The efficacy of selected 'candidate' gene loci in risk algorithms, to improve the predictive accuracy for coronary heart disease, remains to be demonstrated. RECENT FINDINGS: Although many candidate genes for coronary heart disease have been tested, the optimal set of risk genotypes has yet to be identified. There is only a relatively modest risk to be expected in association with any single genotype, published estimates are in the range of 1.12-1.73. Thus the risk associated with any one genotype is modest, but, in combination, selected genotypes may be associated with a clinically significant risk. Since the allele frequency for many of these variants is high, many individuals will carry several 'risk alleles'. A small number of selected single nucleotide polymorphisms should complement the conventional risk factors to identify high-risk individuals in whom correction of 'modifiable risk factors' through lifestyle interventions or medication would be most beneficial. SUMMARY: As our understanding of how genetic variation impacts on common diseases advances, the novel loci identified by genome-wide association scans associated with disease risk will rapidly improve these risk algorithms. PMID- 18388693 TI - Common statistical issues in genome-wide association studies: a review on power, data quality control, genotype calling and population structure. AB - PURPOSE OF REVIEW: Genetic association studies which survey the entire genome have become a common design for uncovering the genetic basis of common diseases, including lipid-related traits. Such studies have identified several novel loci which influence blood lipids. The present review highlights the statistical challenges associated with such large-scale genetic studies and discusses the available methodological strategies for handling these issues. RECENT FINDINGS: The successful analysis of genome-wide data assayed on commercial genotyping arrays depends on careful exploration of the data. Unaccounted sample failures, genotyping errors and population structure can introduce misleading signals that mimic genuine association. Careful interpretation of useful summary statistics and graphical data displays can minimize the extent of false associations that need to be followed up in replication or fine-mapping experiments. SUMMARY: Recently published genome-wide studies are beginning to yield valuable insights into the importance of well designed methodological and statistical techniques for sensible interpretation of the plethora of genetic data generated. PMID- 18388694 TI - Family study designs in the age of genome-wide association studies: experience from the Framingham Heart Study. AB - PURPOSE OF REVIEW: The past year has seen the publication of many genome-wide association studies, most of which are case-control studies. These publications are at the forefront of current research into the examination of genetic effects for numerous diseases, including diabetes, heart disease and cancer. Over the past 25 years the tour de force of genetics research has been in family studies, using segregation, linkage and association analyses. Are these approaches now passe? Here we discuss the role of family studies in modern genetics research, using results from the Framingham Heart Study as examples. RECENT FINDINGS: Family studies permit both linkage and association analyses. Importantly, family based association tests that consider transmission of genetic variants within a family provide important information on the genetic etiology of disease traits and avoid the potential of false-positive findings due to population substructure. SUMMARY: Family-based study designs continue to contribute much to the modern era of genome-wide association studies. PMID- 18388695 TI - 'Deep phenotyping': characterizing populations in the era of genomics and systems biology. AB - PURPOSE OF REVIEW: Large-scale genomic studies establish genotype-phenotype associations, but they use phenotypes that represent current views of disease. There is an opportunity to enhance our understanding of genotype-phenotype associations by extending phenotypes into much greater detail ('deep phenotyping'). RECENT FINDINGS: We should engage in deep phenotyping for the following reasons. First, the current emphasis on clinical outcomes, although necessary for the advancement of clinical medicine, is not sufficient. Second, analytical and biological variance embedded in traditional phenotypes dilutes statistical power and strength of association. Finally, even relatively precise phenotypes may vary in terms of underlying pathophysiology across an individual's life history. Deep phenotyping focuses on the biological relevance of pathways and metabolic flux, increasing the 'granularity' of phenotypes. SUMMARY: Focus on medical phenotypes is critical, but long-term interests require additional studies that illuminate underlying biology. Deep phenotyping is less likely to yield dramatic changes in current medical practice but it offers an opportunity to gain scientific insight in an incremental manner and to make progress in redefining clinical outcomes with greater precision. It is expensive, and debate is needed to determine when and how it should be applied. PMID- 18388696 TI - Why study gene-environment interactions? AB - PURPOSE OF REVIEW: We examine the reasons for investigating gene-environment interactions and address recent reports evaluating interactions between genes and environmental modulators in relation to cardiovascular disease and its common risk factors. RECENT FINDINGS: Studies focusing on smoking, physical activity, and alcohol and coffee consumption are observational and include relatively large sample sizes. They tend to examine single genes, however, and fail to address interactions with other genes and other correlated environmental factors. Studies examining gene-diet interactions include both observational and interventional designs. These studies are smaller, especially those including dietary interventions. Among the reported gene-diet interactions, it is important to highlight the strengthened position of APOA5 as a major gene that is involved in triglyceride metabolism and modulated by dietary factors, and the identification of APOA2 as a modulator of food intake and obesity risk. SUMMARY: The study of gene-environment interactions is an active and much needed area of research. Although technical barriers of genetic studies are rapidly being overcome, inclusion of comprehensive and reliable environmental information represents a significant shortcoming of genetics studies. Progress in this area requires inclusion of larger populations but also more comprehensive, standardized, and precise approaches to capturing environmental information. PMID- 18388697 TI - Bibliography. Current world literature. Genetics and molecular biology. PMID- 18388698 TI - Nutrition and metabolism: alpha-tocopherol to prevent cardiovascular disease- deficiency, dose, delivery, and developing context. PMID- 18388699 TI - Genetics and molecular biology: proprotein convertase subtilisin/kexin type 9 and LDL receptor--an intriguing story. PMID- 18388700 TI - Lipid metabolism: recent progress in defining the contributions of cholesterol transporters to cholesterol efflux in vitro and in vivo. PMID- 18388701 TI - Atherosclerosis: cell biology and lipoproteins. PMID- 18388702 TI - Therapy and clinical trials. PMID- 18388703 TI - Patient care, professionalism, and relations with industry: editorial from journal editors. PMID- 18388704 TI - How safe is the operative treatment of Gartland type 2 supracondylar humerus fractures in children? AB - Whereas operative treatment of supracondylar fractures is now standard of care for Gartland type 3 supracondylar humerus fractures in children, the treatment of type 2 fractures remains somewhat controversial. The purpose of this article was to examine the safety and efficacy of closed reduction and pinning of type 2 supracondylar humerus fractures in children. METHODS: We performed a retrospective review of 189 type 2 supracondylar humerus fractures operatively treated at one tertiary care children's hospital from 2000 to 2006. Data were acquired from a review of radiographs and clinical notes. RESULTS: We found no intraoperative surgical or anesthetic complications in our series. None of our cases lost reduction after closed reduction and percutaneous pinning. There were 4 pin tract infections (2.1%) in our series: 3 were treated with antibiotics, and 1 needed irrigation and debridement in the operating room. This was the only patient who required reoperation for any reason. CONCLUSIONS: In this study, the largest reported series of type 2 supracondylar humerus fractures in children, we found an extremely low rate of complications after closed reduction and percutaneous pinning; secondary operations were also uncommon (0.5%). Our series demonstrates a high probability of satisfactory outcome after operative treatment of type 2 supracondylar fractures compared with previous studies of children treated by closed reduction without pinning. LEVEL OF EVIDENCE: Therapeutic study, level 4 (case series [no or historical control group]). PMID- 18388705 TI - Fractures of the distal humeral metaphyseal-diaphyseal junction in children. AB - BACKGROUND: To our knowledge, there is no report in the English-language literature of metaphyseal-diaphyseal junction fractures of the distal humerus in children. The purpose of this study was to review our experience with this uncommon fracture. METHODS: Between 1998 and 2004, 422 displaced supracondylar humerus fractures underwent operative reduction and fixation at our institution. A retrospective review of medical records and radiographs revealed that 14 (3.3%) of these fractures occurred at the metaphyseal-diaphyseal junction just proximal to the olecranon fossa. In 8 patients, the fracture line was oblique (group A), and in 6 patients, the fracture line was transverse (group B). RESULTS: Average age at the time of fracture was 4.9 years (range, 1.5-10 years). All patients were treated by closed reduction and Kirschner wire fixation and had at least 1 year follow-up. In group A, operative time for reduction and fixation was significantly increased in comparison to the 408 remaining supracondylar humerus fractures. However, the clinical course in group A was uncomplicated, and no loss of fixation at follow-up was noted. The operative time in group B was even longer. These fractures were more problematic as loss of fixation occurred in 5 of the 6 patients, 4 occurring in the sagittal plane. In addition, multiple complications arose in group B including reoperation, cubitus varus, pin migration, and prolonged loss of motion. CONCLUSION: Metaphyseal-diaphyseal junction fractures of the distal humerus in children are rare but can be problematic. The transverse fracture pattern requires additional attention in the operating room with optimal pin fixation. Close postoperative follow-up is necessary. The oblique fracture pattern, while requiring increased time in the operating room for reduction and fixation, is typically stable with the usual fixation used for supracondylar humerus fractures. In summary, metaphyseal diaphyseal junction fractures of the distal humerus are uncommon elbow fractures in children that should be differentiated from the more common supracondylar humerus fracture for optimal outcomes. LEVEL OF EVIDENCE: Therapeutic level 4 (case series). PMID- 18388706 TI - Displaced olecranon fractures in children: a biomechanical analysis of fixation methods. AB - BACKGROUND: Wire and suture methods have been used to stabilize pediatric olecranon fractures. This study (1) compared differences in simulated intraoperative compression during fracture reduction, (2) evaluated articular surface compression during cyclic loading of the tension band, and (3) compared fracture stabilization after cyclic physiologic loading at low/high levels. METHODS: Identical olecranon fractures were created in 10 synthetic ulnae and randomized to suture or wire fixation. Compression after fixation and compression during cyclic loading between 10 and 50 N was measured with a load cell at the articular surface and compared with a 1-way analysis of variance (p < 0.05). Twenty-four fractured synthetic ulna were randomly assigned to wire or suture tension band constructs and low- or high-loading groups. The low-load group (12 ulnae) cycled loading from 3 to 10 N for 100 cycles followed by a failure test. The high-load group (12 ulnae) experienced 10 to 100 N before failure testing. Fracture separation (mm) and failure load (N) were compared using a 2-way analysis of variance (p < 0.05). Ten synthetic ulnae were randomized to wire/suture groups and cyclically loaded between 10 to 50 N while measuring loads across the fracture using a load cell. Correlation data were statistically compared with a Fisher transformation and z test (p < 0.05). RESULTS: Residual compression was statistically greater for wire compared with suture. There was no difference in fracture displacement between groups during low loads. Suture had significantly greater displacement compared with wire at high loads. Failure loads were significantly greater for wire at both load settings. Wires transmitted forces across the joint surface more readily than sutures. CONCLUSIONS: Suture tension bands had lower ultimate failure loads and less compression at the fracture site. However, if low loads are expected or if the fracture is reduced easily, the suture tension band may be an appropriate alternative to wire fixation. CLINICAL RELEVANCE: Perhaps, in small children or when using casts in bigger children, a bioabsorbable suture may be used for fracture stabilization avoiding the need for extensive surgery to remove the fixation material. PMID- 18388707 TI - Elastic stable intramedullary nailing of tibial shaft fractures in children. AB - Most pediatric tibia shaft fractures are amenable to nonoperative treatment with satisfying results, yet surgical stabilization is necessary in certain cases. The purpose of our study was to determine the effectiveness and the complications associated with elastic stable intramedullary nailing in severe pediatric tibial fractures. We retrospectively reviewed 24 tibia shaft fractures in 24 patients that were treated operatively by elastic stable intramedullary nailing between 1997 and 2005 at our institution. Extensive review of all charts and radiographic data was completed at Cincinnati Children's Hospital Medical Center. There were 8 closed and 16 open fractures. The average union time for all tibia fractures was 20.4 weeks. The average union time for closed and open fractures was 21.5 and 20.2 weeks, respectively. Complications include 2 (8%) neurovascular, 2 (8%) infections, 2 (8%) malunions, and 1 (4%) leg-length discrepancy. Although complications do exist, elastic stable intramedullary nailing of pediatric tibia shaft fractures using Nancy nails is an effective treatment option. PMID- 18388708 TI - Biomechanical comparison of four different fixation techniques for pediatric tibial eminence avulsion fractures. AB - BACKGROUND: Several different methods have been used to repair tibial eminence avulsion fractures. It is not clear which is the best stabilization method. The purpose of this study was to compare the biomechanical stability of tibial eminence avulsion fractures using suture, resorbable screw, resorbable nail, and metal screw techniques. METHODS: Sixteen immature bovine knees were dissected leaving just the anterior cruciate ligament. A fracture was created using a curved osteotome, The knees were randomly stabilized with either 2 single-armed #2 Ethibond sutures, 3 bioabsorbable nails, a single resorbable screw, or a single metal screw. Femurs were tested with the knee flexed to 35 degrees to simulate anterior tibial translation. Tests involved loading between 5 N and 150 N for 200 cycles, then a tensile failure test at 0.5 mm/sec. Cyclic fragment deformation, initial fragment stiffness, and failure load were compared using a 1 way analysis of variance (p < 0.05). RESULTS: There were no significant mechanical differences across groups. The variability in performance was much greater for both the suture and resorbable screw repairs. Both sutures and resorbable screw constructs resulted in a deformation that was 1 mm greater than that of the resorbable nails or a metal screw. CONCLUSIONS: Increased fracture separation for sutures and resorbable screw groups indicates a potential loss in reduction during cyclic, physiologic loads. Each group could withstand up to 85 lb of tensile force before failure, but it is unlikely that this force would occur with incidental loads during the early rehabilitation period. CLINICAL RELEVANCE: There was not a clear biomechanical advantage to performing any particular fixation method in this study. This suggests that the surgeon can use their clinical judgment and experience to determine the fixation technique. PMID- 18388709 TI - Mobility in Hurler syndrome. AB - Hurler syndrome is an autosomal recessive metabolic storage disease. Hematopoietic stem cell transplant increases life expectancy, but the effects on associated musculoskeletal abnormalities remains unclear, and long-term data are limited. We detail the follow-up of 23 patients at a mean of 8.5 years after successful hematopoietic stem cell transplant. All patients underwent clinical examination at an annual multidisciplinary clinic. Serial radiological studies were reviewed to assess development and management of hip dysplasia and genu valgum. All patients demonstrated characteristic acetabular dysplasia and failure of ossification of the superolateral femoral head. Eight patients underwent bilateral pelvic and femoral derotation (mean age at surgery, 4.4 years); 4 patients had pelvic osteotomy only. Mean preoperative acetabular angle was 34 degrees. Genu valgum of variable severity due to failure of ossification of the lateral aspect of the proximal tibial metaphysis was observed early, and 6 patients underwent medial epiphyseal stapling, decreasing tibiofemoral angle by a mean of 8 degrees. Clinically, all patients were independently mobile, with restriction of internal hip rotation being the most significant clinical finding. Valgus knees and pronated feet were a typical finding. This cohort represents one of the largest available for study, and ongoing review will clarify the progression of musculoskeletal problems and determine the effectiveness of orthopaedic intervention. PMID- 18388710 TI - Valgus osteotomy: a solution for late presentation of hinge abduction in Legg Calve-Perthes disease. AB - BACKGROUND: This is a study to establish the role of surgery in late presenting hinge abduction following Legg-Calve-Perthes disease. METHODS: A prospective study was carried out involving 15 patients who underwent valgus femoral osteotomy for hinge abduction related to Perthes disease between 1995 and 2000. Surgery was performed by the senior author. There were 2 females and 13 males; in 6 cases, the left hip was affected, and in 9, the right. There were no cases of bilateral Perthes disease. The mean age at the time of surgery was 17 years and 9 months with an age range of 11 years to 32 years and 6 months. All patients were Stulberg grade 3 or 4 preoperatively. RESULTS: The mean follow-up was 78 months (range, 61-95 months). The mean Harris hip score (HHS) preoperatively was 48 with a range of 29 to 75. At a mean of 22 months postoperatively, the HHS improved to a mean of 89 (range, 62-100) points (P < 0.01). There was no subsequent significant alteration in the HHS from the time of initial follow-up to final follow-up at 6.5 years. CONCLUSION: This study illustrates the role of valgus femoral osteotomy in patients presenting late with hinge abduction, demonstrating that the initially early improvement in patient function stabilizes and does not alter at 6 years. LEVEL OF EVIDENCE: Prospective study. Level 1. PMID- 18388711 TI - The incidence of occult dysplasia of the contralateral hip in children with unilateral hip dysplasia. AB - BACKGROUND: The purpose of this study is to determine the incidence of contralateral hip dysplasia in children with unilateral hip dysplasia. METHODS: We reviewed all patients with developmental dysplasia of the hip (DDH) who were evaluated at our institution until skeletal maturity (at least age 13 years). In the contralateral ("normal") hip, radiographic measurements of the acetabular angle >46 degrees, center edge angle <20 degrees, and migration index >20% were considered abnormal. RESULTS: Of the 43 patients in the study, 18 (42%) were diagnosed with bilateral hip dysplasia as an infant or toddler; 8 on the initial visit, and 10 after repeated evaluations. Twenty-five patients had no evidence of contralateral hip dysplasia, even on repeated examinations. Of this group, 10 of 25 (40%) had mild acetabular dysplasia at maturity (mean center edge angle, 31 degrees; acetabular angle, 43 degrees; and migration index, 14%). No child in this group was diagnosed with significant hip dysplasia at skeletal maturity that required treatment to date. DISCUSSION: The results of this study indicated that mild acetabular dysplasia occurs at maturity in the contralateral hip in up to 40% of patients diagnosed with unilateral DDH, even when the contralateral hip was carefully evaluated as a child. The significance of mild acetabular dysplasia is not well defined, but intermittent evaluation of the contralateral hip in children with DDH would seem prudent. LEVEL OF EVIDENCE: IV. PMID- 18388712 TI - An anatomical study of the pediatric intercondylar notch. AB - BACKGROUND: Anterior cruciate ligament reconstructions in skeletally immature patients are becoming more commonplace. Complications of growth disturbance remain a major concern, especially at the distal femoral physis, and are often attributed to technical errors. A review of the literature revealed limited anatomical data of the skeletally immature intercondylar notch to guide surgical technique. METHODS: We studied 103 preserved femora aged 3 to 20 years, with 33 of these aged 7 to 15 years. The distance between the "resident's ridge" (lateral intercondylar ridge) and the "over-the-top" position (OTP) was measured at the 11 , 10-, and 9-o'clock positions in the right femora, and the 1-, 2-, and 3-o'clock positions in the left femora. The angles between the femoral surface, distal femoral physis, and femoral shaft were measured in the coronal and sagittal planes using a goniometer. RESULTS: Femora in the 13- to 15-year-old subgroup tended to have a more clearly defined resident's ridge than femora in the younger subgroups. The space between resident's ridge and the OTP was on average greater than 8 mm at all 3 positions in the 13- to 15-year-old subgroup, and greater than 7 mm at all 3 positions in the 10- to 12-year-old subgroup, with more space available with a more peripheral starting point. The average angles between the femoral surface and physis in the sagittal and coronal planes were 47 degrees and 36 degrees in the 13- to 15-year-old subgroup, and 58 degrees and 28 degrees in the 10- to 12-year-old subgroup. The average angles between the distal femoral shaft and physis in the sagittal and coronal planes were -9 degrees and 7 degrees in the 13- to 15-year-old subgroup, and -6 degrees and 7 degrees in the 10- to 12 year-old subgroup. Analysis of females versus males demonstrated larger distances between resident's ridge and the OTP in males, but no differences in the angular measurements. CONCLUSIONS: In younger patients, the resident's ridge is less commonly present and less clearly defined. The femoral tunnel can usually be drilled behind the resident's ridge at the 11-o'clock/1-o'clock position, although in females, the ridge is more likely to be included in the drill hole. In both males and females, the tunnel should be aimed posterior and medial with respect to the perpendicular of the femoral surface and anterior and medial with respect to the shaft to drill through the physis with less obliquity, and to provide a tunnel deep enough to avoid spanning the physis with bone or hardware. PMID- 18388713 TI - Ligamentous laxity of the knee during childhood and adolescence. AB - BACKGROUND: There is little age- and sex-specific objective data concerning anterior knee translation in school-aged children. We aimed to establish age- and sex-specific baseline data for anterior knee translation of the knee in girls and in boys in grades 5 to 12. We also investigated whether a correlation exists between anterior knee translation and global joint laxity in girls in grades 5 to 12. METHODS: A sample of convenience from a local school was used. Fifteen boys and 15 girls were randomly chosen from each grade, 5 to 12, to undergo anterior translation measurements with the KT2000 knee ligament arthrometer at 15, 20, and 30 lb of force. In addition, Beighton scores and bone age were determined. RESULTS: Anterior knee translation decreased with age in both girls and boys. Girls had greater anterior knee translation and end-point compliance than age matched boys. In girls, there was a positive correlation between increased global joint laxity and increased anterior knee translation at 20 and 30 lb. However, hyperextension of the knee by itself was not a predictor of greater anterior translation. CONCLUSIONS: Anterior translation of the knee showed age and sex variation in school-aged children. These data are important in establishing a foundation of baseline data to understand sex differences in ligament development and function and to help guide injury prevention strategies. LEVEL OF EVIDENCE: Level II diagnostic study. PMID- 18388714 TI - Hemiepiphysiodesis for the correction of angular deformity about the knee. AB - BACKGROUND: It is generally accepted that the presence of angular deformity about the knee that persists into the preadolescent years will not correct spontaneously. The goal of this study was to review our experience with hemiepiphysiodesis and physeal stapling for the correction of angular deformity to establish the efficacy of these procedures and to determine their indications and the rate of correction. METHODS: A retrospective review of 48 patients undergoing hemiepiphysiodesis or physeal stapling about the knee was performed. Patients were included if they had at least 2 years of documented follow-up without an osteotomy. RESULTS: The average follow-up was 30.4 months (range, 24 52 months). Genu varum was identified in 33 patients, genu valgum in 14 patients, and a windswept deformity in 1. Of the 52 limbs with genu varum, 35 were secondary to Blount disease. The group with genu varum unrelated to Blount disease presented a change in the mechanical axis from a mean of -32 degrees at surgery to a mean of - 13 degrees. The patients with Blount disease presented a change in the mechanical axis from a mean of -19 degrees at surgery to a mean of 16 degrees. The group with genu valgum presented a change in the mechanical axis from a mean of 20 degrees at surgery to a mean of 2 degrees. CONCLUSIONS: Hemiepiphysiodesis is an effective means for correcting angular deformity about the knee in skeletally immature patients with both genu valgum and genu varum, so long as the genu varum is unrelated to Blount disease. Hemiepiphysiodesis is effective in preventing the progression of deformity in patients with Blount disease. LEVEL OF EVIDENCE: Therapeutic-Level IV. PMID- 18388715 TI - Analysis of the pediatric outcomes data collection instrument in ambulatory children with cerebral palsy using confirmatory factor analysis and item response theory methods. AB - BACKGROUND: Traditional use of the Pediatric Outcomes Data Collection Instrument (PODCI) assumes that all items have the same structure, are measuring the intended constructs, and assess the right levels of function to show change after orthopaedic or neurological intervention. Item response theory (IRT) methods can statistically account for inherent differences in PODCI item characteristics and thus reveal attributes of the measure important to effectiveness research. Our study uses IRT methods to determine whether PODCI items fit the projected dimensional structure of the PODCI, assess function on each dimension at the right level for a population of ambulatory children with cerebral palsy (CP), and reveal changes after intervention in this population. METHODS: Proxy-reported PODCI questionnaires for 570 ambulatory children with CP were randomly divided into 2 groups for model creation and model testing using exploratory and then confirmatory factor analysis. The resulting model was compared with the projected dimensional structure, tested for fit of individual items, and examined for gaps and ceiling effects. Response changes at 1 year were compared between those with (n = 91) and without (n = 284) surgical intervention using paired t tests. RESULTS: Factor analysis reduced the projected dimensions from 5 to 4 for this population, resulting in dimensions for mobility, upper extremity function (UEF), comfort and general health, and self-worth. All but 3 items fit their respective dimensions; ceiling effects were noted in 3 dimensions. Responses showed changes in the comfort and general health, mobility, and UEF dimensions in those who had surgery; in those children who did not have surgery, only the UEF responses changed. CONCLUSIONS: The PODCI can show change after intervention when data are analyzed using IRT methods. Ceiling effects in 3 dimensions may limit the amount of change the PODCI can show in a population of ambulatory children with CP. LEVEL OF EVIDENCE: Level II. This was a retrospective investigation of a diagnostic tool, the PODCI, using a randomized cross-sectional design for model development, and a case-control design to assess sensitivity to change. PMID- 18388717 TI - Effect of intrathecal baclofen on dystonia in children with cerebral palsy and the use of functional scales. AB - BACKGROUND: The literature available shows that spasticity in patients with cerebral palsy (CP) is reduced by intrathecal baclofen (ITB) treatment, and various studies suggest that this treatment can also be used in dystonic patients. The aim of the present study was to evaluate dystonia treated with ITB in children with CP. METHODS: Data of 19 patients affected by dystonia as an outcome of infant CP in patients belonging to level V of Gross Motor Function Classification System treated with ITB were collected. The mean age at implant is 8.49 years.The patients are assessed using Barry-Albright Scale and Burke-Fahn Marsden Scale before treatment and at 3, 6, and 12 months postimplant. RESULTS: The results showed a statistically significant improvement (P < 0.001) in the total scores of the 2 scales after just 3 months of treatment, an improvement that was maintained for the 1-year period of follow-up. The results also revealed a reduction in dystonias, an improvement in posture, and an easing of the task of the caregivers in managing the patient as a result of treatment with ITB. CONCLUSIONS: In patients belonging to level V of Gross Motor Function Classification System and treated with ITB, a decrease in frequency and severity of dystonia is observed. This improvement eases caregiver in patient management. LEVEL OF EVIDENCE: Therapeutic Level IV. PMID- 18388716 TI - Concurrent and discriminant validity of Spanish language instruments for measuring functional health status. AB - BACKGROUND: Questionnaires translated into languages other than English are often not validated to the same extent as the English versions. This study examined the concurrent and discriminant validity of selected domains related to physical function from Spanish language versions of the Child Health Questionnaire (CHQ), Pediatric Outcomes Data Collection Instrument (PODCI), and Pediatric Evaluation and Disability Inventory (PEDI). METHODS: Concurrent validity was examined in 93 children with cerebral palsy by correlating questionnaire domain scores with Gross Motor Function Measure and Gillette Functional Assessment Questionnaire walking scale scores. Discriminant validity with respect to Gross Motor Function Classification System (GMFCS) level was examined using analysis of variance and nonparametric discriminant analysis. RESULTS: Concurrent validity was demonstrated for 3 domains from the PEDI (Mobility functional skills, tau = 0.62; Mobility caregiver assistance, tau = 0.46-0.55; and Self-care functional skills, tau = 0.30-0.36), 3 domains from the PODCI (Sports and physical function, tau = 0.48-0.51; Transfer and basic mobility, tau = 0.48-0.51; and Upper extremity physical function, tau = 0.28), and 1 domain from the CHQ (Physical function, tau = 0.31-0.36). Discriminant validity was demonstrated for the same domains based on significant decreases in domain scores with increasing GMFCS level. Discriminant validity was highest for the PODCI, which correctly classified 98% (91/93) of subjects into the correct GMFCS level when all 3 domains were considered. CONCLUSIONS: For the first time, concurrent validity and discriminant validity have been demonstrated for the physical function domains of Spanish language versions of the PODCI, PEDI, and CHQ questionnaires. PODCI and PEDI had the highest concurrent validity, and PODCI had the best discriminant ability. CLINICAL RELEVANCE: It is important to examine the validity of instruments when they have been translated from English into other languages. This importance will only increase as the population of non-English-speaking patients expands. PMID- 18388718 TI - Postural characteristics of adolescents with idiopathic scoliosis. AB - BACKGROUND: The objectives of this study were to compare the postural characteristics of idiopathic scoliosis (IS) patients with different types of spinal curvature and to compare a motion capture and a sequential digitization technique to estimate the postural characteristics of the IS patients. METHODS: A total of 57 IS patients underwent a radiological, clinical, and postural geometric evaluations in an upright standing position as part of their regular follow-up. The posteroanterior radiograph of the trunk was used to measure the amplitude of spinal curvature. The postural evaluation was performed using a motion capture and sequential digitization technique providing the necessary 3 dimensional positions of anatomical landmarks. These landmarks were used to calculate postural parameters defining the position and orientation of the pelvis, shoulders, and shoulder blades. These measurements included lateral shift and angular measures of rotation and tilt. RESULTS: Significant differences in the frontal and transverse planes were found between the right thoracic (RTh), left thoracolumbar, and RTh-left lumbar (RThLL) patients. These characteristics were most prominent in the transverse plane for RTh patients, in the frontal plane for left thoracolumbar patients, and in the frontal and transverse planes for the RThLL lumbar patients. A strong positive intraclass correlation was also found between the parameters estimated with the motion capture system and with the sequential digitization system. CONCLUSIONS: Unique postural characteristics that are related to the type of the spinal curvature are evident in adolescents who have IS. Further work is necessary to assess how these measures may be used to monitor the progression of the spinal deformity. CLINICAL RELEVANCE: Quantifying the postural alignment of IS patients using surface anthropometric landmarks provides an opportunity to characterize the unique postural attributes that accompany each type of spinal curvature. PMID- 18388719 TI - Intravertebral deformation in idiopathic scoliosis: a transverse plane computer tomographic study. AB - BACKGROUND: The scoliotic vertebrae are submitted to (1) the displacement in the 3-dimensional space and (2) the bone remodeling, which results in a 3-dimensional intrinsic vertebral deformation. Both phenomena are most expressed inside the apical zone of the curve and can be measured in a computer tomographic (CT) scan. A comparative study of CT thoracic scans in scoliotic and normal children was performed to provide a better description of the altered anatomy with respect to patomechanism of scoliosis. METHODS: Twenty-three scoliotic girls, aged 14.3 +/- 2.1 years, a case of a right thoracic curve with a Cobb angle of 60.6 +/- 19.3 degrees, and 24 controls, free of spinal deformity, sex- and age-matched, underwent CT examination of the thorax at the level of Th8 to Th9 vertebra. The rotation angle of the apical vertebra and the sagittal to coronal rib cage diameters ratio were measured. The intravertebral deformation was assessed by measuring the angles between the axis of the whole vertebra and the axes of the spinous or transverse processes. RESULTS: The ratio of sagittal to coronal chest diameter was reduced in scoliosis patients (P < 0.001) and correlated with the Cobb angle. The angle between the axis of vertebra and the spinous process increased (P = 0.008), and its value was positively correlated with the rotation angle of the vertebra (r = 0.78, P < 0.05); however, the rotation was oriented clockwise, whereas the spinous process deviation was counterclockwise. The angle between the spinous and the transverse process revealed greater values on the concave side (P < 0.001), whereas the transverse processes were not deviated from the axis of vertebra (P = 0.469). CONCLUSIONS: A constant pattern, previously not described, of the alteration of morphology of the apical vertebra due to the intravertebral bone remodeling was identified. The intravertebral deformation accompanied the displacement of the vertebra with a linear correlation; however, the 2 phenomena were developing in the opposite directions. CLINICAL RELEVANCE: Level III, cross-sectional study. PMID- 18388720 TI - Diagnostic modalities for the evaluation of pediatric back pain: a prospective study. AB - The commonly taught premise that pediatric back pain frequently has an underlying diagnosis has been recently challenged. Previous studies have suggested that up to 84% of children with low back pain have associated serious diagnoses. Children with back pain, therefore, have frequently undergone exhaustive diagnostic testing. There have been few prospective studies, however, about the diagnosis rate and appropriate diagnostic methods for back pain in children. This study prospectively examines the rate of diagnosis for pediatric back pain and the value of various diagnostic studies for this problem. METHODS: All patients presenting to our institution with a chief complaint of back pain were evaluated for the study. Inclusion criteria consisted of age younger than 18 years, no previous back surgery, no previous diagnosis given, and duration of pain longer than 3 months. Seventy-three patients were enrolled in the study, and an algorithm was created for diagnostic evaluation. The algorithm incorporated commonly used diagnostic techniques including radiographs, magnetic resonance imaging, computed tomography, bone scan, and laboratory studies. The end point was considered to be either (1) a definitive diagnosis or (2) no diagnosis and no symptomatic or clinical changes during a 2-year period. RESULTS: Fifty-seven patients (78.1%) ended with no diagnosis. Of the remaining 16, 9 were diagnosed with spondylolysis with or without spondylolisthesis. Three other patients had abnormal laboratory values but no definitive diagnosis. Other diagnoses included Scheuermann disease (n = 2), osteoid osteoma (n = 1), and a herniated disk (n = 1). CONCLUSIONS: This investigation is the largest prospective study of diagnostic modalities in pediatric back pain to date. Contrary to most of the previously published data, most of our patients ended the study with no definitive diagnosis. In addition, the most of the diagnoses were made at initial physical examination or via initial plain radiographs. No diagnoses were missed using our algorithm. These results suggest that pediatric back pain frequently does not carry a definitive diagnosis and that exhaustive diagnostic protocols may not be necessary for this problem. LEVEL OF EVIDENCE: Prospective study; Level 2 clinical evidence. PMID- 18388721 TI - Friedreich's ataxia and scoliosis: the experience at two institutions. AB - PURPOSE: Friedreich's ataxia is a genetically transmitted, progressive spinocerebellar degenerative disease characterized by ataxia. The purpose of this study is to evaluate the demographics, progression, nonoperative, and operative treatment of spinal deformities in patients with Friedreich's ataxia at 2 tertiary pediatric orthopaedic hospitals. METHODS: After institutional review board approval, chart review of Friedreich's ataxia patients identified those having scoliosis. Demographic data, length of follow-up, brace treatment, operative treatment, and complications were determined. Radiographic review was also performed. RESULTS: Seventy-seven patients were identified as having Friedreich's ataxia, of which 49 (63%) were diagnosed with scoliosis. Twenty seven were male; 22 were female. Mean age at diagnosis of scoliosis was 12.8 years (4.9-20 years). Mean follow-up was 3.7 years (0-13 years). There were 16 (33%) double major curves, with 8 (22%) of the thoracic curves being left sided. Hyperkyphosis was present in 12 (24.5%).Twenty-four (49%) of patients progressed > or =6 degrees. Using a chi-square analysis, there was no association, with a curve magnitude of 10 degrees before the age of 10 years and progression of the curve (P = 0.4386). Ten (20%) patients were treated in braces, with average progression in brace of 15 (0-44) degrees. Sixteen (33%) patients were treated with spinal fusion (15 posterior spinal fusion and 1 anterior spinal fusion). Thirteen (81%) of 16 patients who underwent operative intervention were wheelchair dependent. Somatosensory evoked potentials monitoring was attempted in 11 patients but was effective in only 1. Immediate postoperative correction averaged 49% in the thoracic spine (24%-87%) and 51% in the lumbar spine (26% 82%). This correction decreased to 39% in the thoracic (-22% to 85 %) and 30% in the lumbar spine (-35% to 82%) at final follow-up. The average postoperative follow-up was 3.6 years (2-6.5). One patient (6.2%) developed an infection and was the only patient who underwent reoperation. CONCLUSIONS: Scoliosis in Friedreich's ataxia is common (63%). Curve patterns are variable and do not necessarily resemble idiopathic curves. Although few patients were braced, results were poor. Fusion using modern segmental constructs was effective in creating substantial intraoperative correction and maintaining correction postoperatively. SSEP monitoring was usually ineffective, so preparation for a wake-up test is recommended. SIGNIFICANCE: Patients with Friedreich's ataxia need to be carefully screened for scoliosis and counseled about the high rate of surgical fusion. Using modern implants, correction can be achieved and maintained. PMID- 18388722 TI - Health-related quality of life in children with thoracic insufficiency syndrome. AB - BACKGROUND: The traditional techniques to treat thoracic insufficiency syndrome (TIS) are not able to stabilize or improve chest wall size or pulmonary function while allowing spine growth. To this end, vertical expandable prosthetic titanium rib (VEPTR) was specifically designed to treat TIS by allowing growth of the thoracic cavity and control/correction of spine deformity. The purpose of this study was to determine quality of life (QOL) of children with TIS and its impact on their parents before and after implantation of the VEPTR and also compare these results to those of healthy children. METHODS: As part of the original multicenter evaluation of the VEPTR, a Child Health Questionnaire (CHQ) was collected preoperatively on 45 patients who were subsequently treated with expansion thoracoplasty using the VEPTR. The average age was 8.2 +/- 2.6 years, and the parent form of the CHQ was filled out by the primary caretaker. Patients were divided into 3 diagnostic categories: rib fusion (n = 15), hypoplastic thorax syndromes (n = 17), and progressive spinal deformity (n = 13). RESULTS: There were significant differences between the study patients and healthy children in physical domains. Compared with parents of healthy children, parents of children with TIS experienced more limitations on their time and emotional lives due to their children's health problems. There were no significant differences in CHQ before and after the surgery except for a significant decrease in the self-esteem among a subgroup of patients with hypoplastic thorax syndromes. There were no significant differences in postoperative QOL between patients who had VEPTR-related complications and patients who did not have the complications. CONCLUSIONS: The children with TIS had lower physical scores and higher caregiver burden scores than healthy children. However, the scores in psychosocial domains were similar to those in healthy children. Our study demonstrated that QOL of children and burden of care in their parents remained the same after VEPTR instrumentation. Children's QOL seemed to be not affected by whether they had VEPTR-related complications or not. PMID- 18388723 TI - Efficacy of spinal instrumentation and fusion in the prevention of postlaminectomy spinal deformity in children with intramedullary spinal cord tumors. AB - Postlaminectomy spinal deformity is a frequent problem after resection of a pediatric spinal cord tumor. However, the use of spinal fusion done at the time of resection in preventing the development of deformity is unknown. The purpose of our study was to assess the effectiveness of single-stage laminectomy, spinal cord decompression, and fusion for the prevention of postlaminectomy spinal deformity in children with intramedullary spinal cord tumors. Clinical charts and radiographs of 33 children with spinal cord tumors were reviewed for treatment description, preoperative spinal alignment, postoperative spinal alignment, and subsequent medical and surgical treatment. Spinal deformity was graded by predefined criteria. Significant spinal deformity developed in 10 of 17 children with laminectomy only, 3 of 4 children with laminoplasty, and 3 of 12 children with spinal fusion (2/6 with instrumentation, 1/6 with in situ fusion). Excluding patients that progressed to paraplegia, 9 of 16 patients treated with resection without fusion compared with 1 of 10 patients treated with resection and fusion developed a spinal deformity (P = 0.04). Among skeletally mature children that did not progress to paraplegia, 9 of 15 treated with resection without fusion compared with 1 of 7 patients treated with resection and fusion developed a deformity (P = 0.05). Removal of greater than 4 laminae (P = 0.03) was found to be associated with the development of postresection spinal deformity. PMID- 18388724 TI - Treatment of clubfoot with the Ponseti method: a comparison of casting materials. AB - BACKGROUND: Popular initial treatment for congenital clubfoot includes the use of serial manipulations and casting as described by Ponseti et al. Plaster of Paris and semirigid fiberglass are 2 materials commonly used for casting. To our knowledge, no study to date has compared the clinical results of these 2 materials. The objective of this randomized prospective study was to compare the effectiveness of these materials in the initial management of clubfoot. METHODS: All clubfeet presenting to the 2 senior authors' outpatient clinics over a 15 month period were offered enrollment. Patients were randomly assigned for treatment with either plaster or semirigid fiberglass casts. The severity of the clubfoot deformity was documented using the scoring system devised by Dimeglio et al. Serial casts were applied according to the technique described by Ponseti et al. At the completion of nonsurgical treatment, the final clubfoot severity was documented. RESULTS: A total of 42 clubfeet in 34 patients were enrolled in the study. After exclusion of 3 patients, 13 patients (16 feet) received fiberglass, and 18 patients (23 feet) received plaster casts. The mean baseline severity scores of the 2 groups were not significantly different. The mean final severity score was significantly higher in the feet treated with fiberglass than those treated with plaster (6.4 vs 4.1; P = 0.037). There was a trend toward higher scores for cast tolerance, durability, and parent satisfaction in the fiberglass group, but this did not reach significance. CONCLUSIONS: This study supports the use of plaster casting with the Ponseti technique. The use of plaster casts resulted in a statistically lower Dimeglio-Bensahel score at the completion of serial casting. There was a trend toward higher patient satisfaction in the fiberglass-treated group. Whether this difference has an effect on long-term outcomes and recurrence remains to be studied. LEVEL OF EVIDENCE: Level II. Nonblinded randomized controlled prospective study. PMID- 18388725 TI - Dynamic pedobarography for children: use of the center of pressure progression. AB - BACKGROUND: Although pedobarography has been widely used in quantitative clinical gait analysis for children, the collection, processing, analysis, and interpretation of the data vary widely. In most cases in children, foot dysfunction during gait is primarily a consequence of skeletal segmental malalignment, which can be characterized by the location and duration of the center of pressure progression (COPP) relative to the foot. This study determines the validity and reliability of a technique using the COPP and establishes a normative database for the COPP in children. STUDY DESIGN: Prospective case series to evaluate a diagnostic test. METHODS: Simultaneous pedobarograph and kinematic data collection was performed on 23 children (46 feet) who were neurologically healthy. The validity of the COPP technique was determined by comparing the pedobarograph- and kinematic-based determinations of the orientation of the longitudinal (or long) axis of the foot, an essential component of the COPP approach. Intrarater and interrater reliability for the pedobarograph-based technique were determined by comparing repeated measures of the long axis of the foot from 4 analysts. Normative data for the location and duration of the COPP were generated from this cohort of neurologically healthy children. RESULTS: The mean difference for the long axis of the foot between the pedobarograph- and kinematic-based methods was 2.3 degrees (P < 0.001). The mean difference between first and second determinations of the long axis of the foot by the same analyst was 1.0 degrees (P < 0.001; correlation coefficient, 0.975). The mean difference between the 4 analysts' determinations of the long axis of the foot was 1.9 degrees (P < 0.001; correlation coefficient, 0.969). The normal COPP is located under the heel segment for 23.7% of stance, under the midfoot segment for 28.7% of stance, and under the forefoot segment for 47.5% of stance. CONCLUSIONS: This study establishes clinically acceptable validity and reliability for the pedobarograph COPP technique and determines the location and duration of the COPP in a cohort of neurologically healthy children. CLINICAL RELEVANCE: This standardized approach to the determination of foot loading patterns, based upon normative data, should facilitate the characterization of abnormal foot loading patterns, clinical decision making, and the assessment of outcome after a variety of interventions. PMID- 18388726 TI - Comparison of pediatric outcomes data collection instrument scores and range of motion before and after shoulder tendon transfers for children with brachial plexus birth palsy. AB - BACKGROUND: Children with brachial plexus birth palsy (BPBP) may undergo shoulder external rotation tendon transfers (ERTT) to improve function. In terms of outcome measurements, and according to the World Health Organization International Classification of Functioning, Disability, and Health model, ERTT reduces impairment as measured by range of motion (ROM), but has not been shown to improve activity and participation. Furthermore, correlation between these outcome measures has not been established. The Pediatric Outcomes Data Collection Instrument (PODCI) is a well-validated musculoskeletal health questionnaire that addresses activity and participation components of function. The aim of this study is to determine whether PODCI scores improve after ERTT, and whether this improvement correlates with improvement in ROM. METHODS: A total of 23 children with a mean age of 6.3 years (range, 4.4-12.8 years) with BPBP and standard indications for ERTT underwent preoperative and 1-year postoperative shoulder ROM measurements, and parental completion of the PODCI. Change in ROM was compared with change in PODCI scores to determine if these were correlated. RESULTS: Average range of active shoulder abduction improved 35 degrees (P < 0.001), and average range of active external rotation improved 41 degrees (P < 0.001). The PODCI scores for Upper Extremity Function, Sports Function, and Global Function improved (12 points [P < 0.001], 4 points [P = 0.04], and 6 points [P = 0.001], respectively). Improvement in ROM did not correlate with improvement in PODCI scores. However, postoperative peak active abduction correlated strongly with postoperative PODCI scores for Upper Extremity Function, and Global Function (rs = 0.712 [P < 0.001], rs = 0.735 [P < 0.001], respectively), and moderately with Transfers and Basic Mobility and Sports Function scores (rs = 0.496 [P=0.016], rs = 0.449 [P = 0.032], respectively). CONCLUSIONS: For children with BPBP, ERTT is associated with reduced impairment and improved activity and participation. Maximum postoperative abduction is positively associated with PODCI scores, but change in ROM is not. Further study is needed to determine if ceiling effects or other factors account for the lack of correlation between these outcome measures. LEVEL OF EVIDENCE: Level of evidence IV, case series. PMID- 18388727 TI - Image-guided laser ablation of osteoid osteoma in pediatric patients. AB - BACKGROUND: We aimed to evaluate the effectiveness of image-guided laser ablation for the treatment of osteoid osteomas in pediatric patients. METHODS: From June 1994 to June 2006, 68 patients aged 3 to 18 years (mean age, 12.1 +/- 4.2 years) with a diagnosis of osteoid osteoma underwent laser ablation in our department. Among them, 3 patients had prior unsuccessful attempt of surgical resection. Procedures were performed under spinal or general anesthesia with combined computed tomographic and fluoroscopic guidance. An 18-gauge needle was placed in the nidus after bone drilling when required, and an optical fiber was inserted through it. An average energy of 1200 J was delivered using an infrared diode laser generator. Patients were discharged the next day and followed up with assessment of pain, medication intakes, and potential complications. Clinical success was defined by complete pain relief without need for medication intake. RESULTS: Technical success was achieved in all but 1 of our first patients, which was subsequently treated by percutaneous resection. Overall clinical success was 98%. Primary clinical success was obtained in 60 patients (88%). In 7 patients, recurrence of symptoms at 4 to 27 months from the initial procedure was managed by repeat laser ablation, resulting in secondary success (10%). No complications related to the procedure were observed. Follow-up ranged 16 to 146 months (mean, 83 months). CONCLUSIONS: Currently, image-guided laser ablation is a safe and effective procedure that can be proposed as the mainstay treatment of osteoid osteomas in pediatrics. LEVEL OF EVIDENCE: Therapeutic case series study level IV. PMID- 18388728 TI - Recreational terminal devices for children with upper extremity amputations. AB - Children with upper extremity amputations are particularly challenged in sports and other recreational activities. Many find that a prosthesis is of no benefit in their routine daily activities. Although a number of recreational terminal devices are available for prosthetic modification, it is unclear how effective they are at improving a child's performance in sports or hobbies. METHODS: We evaluated the usefulness of 15 recreational terminal devices for upper extremity prostheses in 11 children by chart review and patient survey. All patients had functional elbows, and all but one was a unilateral amputee. All patients were instructed in their use by a prosthetist, and 3 patients received occupational therapy. RESULTS: Nine of 15 recreational terminal devices prescribed improved performance. However, they primarily were the adaptations for weight lifting and violin bows, where their use was obligatory for participation. The wholesale costs for the terminal devices ranged from $120 to $957, and many patients also required the construction of a prosthesis just for the activity. Only 4 of the 9 successful recreational terminal devices were still in use at average follow-up of 3.9 years because patients had lost interest in the activity or had designed something that worked better. CONCLUSIONS: Recreational terminal devices available for weight lifting and violin bows uniformly improved the child's ability to pursue those activities. These prosthetic adaptations are expensive and must be weighed individually by the family in light of children's fleeting interest in individual hobbies. LEVELS OF EVIDENCE: Level IV. PMID- 18388729 TI - Nucleus accumbens activation mediates the influence of reward cues on financial risk taking. AB - In functional magnetic resonance imaging research, nucleus accumbens (NAcc) activation spontaneously increases before financial risk taking. As anticipation of diverse rewards can increase NAcc activation, even incidental reward cues may influence financial risk taking. Using event-related functional magnetic resonance imaging, we predicted and found that anticipation of viewing rewarding stimuli (erotic pictures for 15 heterosexual men) increased financial risk taking, and that this effect was partially mediated by increases in NAcc activation. These results are consistent with the notion that incidental reward cues influence financial risk taking by altering anticipatory affect, and so identify a neuropsychological mechanism that may underlie effective emotional appeals in financial, marketing, and political domains. PMID- 18388730 TI - Monoamine oxidase A-uVNTR genotype affects limbic brain activity in response to affective facial stimuli. AB - Monoamine oxidase A (MAOA) enzymatically degrades biogenic amines such as norepinephrine, dopamine, and serotonin, and plays a key role in the regulation of their neurotransmission. Allelic variations at the MAOA locus have been implicated in the neurobiology of aggression and impulsivity. We investigated the possible relationship between the MAOA-upstream variable number of tandem repeats (uVNTR) polymorphism and brain responses to negative facial stimuli, using functional magnetic resonance imaging (fMRI). We found a significant association between a low activity allele of MAOA-uVNTR and neural activation to negative facial stimuli. In the sad condition, participants with the low activity allele showed greater brain activity in the left amygdala. In the angry condition, participants with the low activity allele showed greater brain activity in the right anterior cingulate cortex and hippocampus. Our results suggest that MAOA uVNTR polymorphism can affect activation of limbic regions, elicited by negative emotional stimuli. PMID- 18388731 TI - Rit contributes to neurite outgrowth triggered by the alpha subunit of Go. AB - Heterotrimeric GTP-binding protein transduce signals initiated by a variety of hormones and neurotransmitters. Go, a member of the Go/Gi family, is the most abundant heterotrimeric GTP-binding protein in nervous tissues and has been implicated in neuronal differentiation. The mechanism by which Go modulates neuronal differentiation has not been, however, fully elucidated. Here, we identified small GTPase Rit as an interacting partner of the alpha-subunit of Go (Goalpha). The biochemical characterizations of Goalpha::Rit interaction revealed that Rit is a candidate downstream effector for Goalpha. Furthermore, dominant negative Rit inhibited Goalpha-induced neurite outgrowth and Erk phosphorylation in Neuro2a cells. These results suggest that Rit may be involved in the signaling pathway for Goalpha-mediated neuronal differentiation. PMID- 18388732 TI - Individual differences in disgust imagery: a functional magnetic resonance imaging study. AB - Functional magnetic resonance imaging studies have examined neural correlates of disgust imagery, but have never taken into account the moderating effects of personality traits. Twenty-four women first viewed and subsequently visualized pictures with disgust-inducing and happiness-inducing content. Relative to the picture perception, disgust, and happiness imagery provoked activation of the insula, anterior cingulate cortex, and parietal cortex. Trait disgust was negatively correlated with localized brain activation (e.g. insula, amygdala, parietal cortex, anterior cingulate cortex) during disgust imagery. This study provides first evidence that disgust propensity is associated with brain activation during imagery of repulsive scenes. PMID- 18388733 TI - Che-1 enhances cyclin-dependent kinase 5 expression and interacts with the active kinase-complex. AB - Che-1 is a nuclear protein involved in the regulation of gene transcription and cell proliferation. It has also been shown to localize to the cytoplasm of postmitotic neuronal cells, where it is able to interact with the microtubule associated protein tau. Cyclin-dependent kinase 5 (Cdk5) is a postmitotic proline directed serine/threonine kinase that hyperphosphorylates tau under pathological conditions. We observed that Che-1 overexpression induces Cdk5 expression both at the mRNA and protein levels. Furthermore, we show that Che-1 directly interacts with Cdk5 protein in vivo. Cdk5/Che-1 complex formation does not compete with Cdk5/p35 interaction, thus Che-1 is able to bind the active kinase complex. Finally, we demonstrated that Che-1 is itself a Cdk5 substrate. PMID- 18388734 TI - Effect of menstrual cycle on resting brain metabolism in female rhesus monkeys. AB - Little is known about the effects of the menstrual cycle on brain activity in primates. Here, we use 18F-fluorodeoxyglucose positron emission tomography to monitor changes in resting brain glucose metabolism across the menstrual cycle in female rhesus monkeys. Results showed greater activity in right lateral orbitofrontal cortex, a region involved in processing negatively valenced emotional stimuli, in the follicular compared with luteal phase. Estradiol levels were negatively correlated with activity in cortical and brainstem regions involved in emotional processing, and positively correlated with activity in areas involved in cognitive control and emotion regulation. In summary, the data suggest that in primates, fluctuations of ovarian hormones across the menstrual cycle influence activity in brain areas involved in emotion and its regulation. PMID- 18388735 TI - Aging hippocampus and amygdala. AB - Earlier studies suggest that the anterior hippocampus may show resilience to age associated volume loss. This study compared high-resolution magnetic resonance images obtained from younger (n=28; age range: 22-50 years) and older (n=39; age range: 65-84 years) healthy right-handed individuals to determine whether age related volume changes varied between the hippocampal head, body and tail. Volumetric reductions were progressively more severe from hippocampal head to tail. Amygdala volume differences were intermediate in size. Although limited by the cross-sectional design, these data suggest that hippocampal subregions show a gradient of volume reduction in healthy aging that contrasts with the preferential reduction of anterior hippocampal volumes in Alzheimer's and Parkinson's diseases. PMID- 18388736 TI - Chronic exposure to low-intensity magnetic field improves acquisition and maintenance of memory. AB - Although past research has suggested that acute exposure to extremely low frequency magnetic field (ELF MF) impairs learning and memory function, data on chronic exposure remain scarce. In this study, we examined the changes in spatial learning and memory by the Morris water maze test after 4 weeks of daily exposure of rats to a 50-Hz magnetic field of 2 mT for either 1 or 4 h. We found that chronic exposure to ELF MF reduced the latency to find the hidden platform and improved long-term memory of former location of platform without affecting the short-term memory and motor activity. These findings for the first time indicate that chronic exposure to ELF MF exerts a positive effect on the acquisition and maintenance of spatial memory. PMID- 18388737 TI - Irrelevant visual stimuli improve auditory task performance. AB - Multisensory behavioral benefits generally occur when one modality provides improved or disambiguating information to another. Here, we show benefits when no information is apparently provided. Participants performed an auditory frequency discrimination task in which auditory stimuli were paired with uninformative visual stimuli. Visual-auditory stimulus onset asynchrony was varied between -10 ms (sound first) to 80 ms without compromising perceptual simultaneity. In most stimulus onset asynchrony conditions, response times to audiovisual pairs were significantly shorter than auditory-alone controls. This suggests a general processing advantage for multisensory stimuli over unisensory stimuli, even when only one modality is informative. Response times were shortest with an auditory delay of 65 ms, indicating an audiovisual 'perceptual optimum' that may be related to processing simultaneity. PMID- 18388738 TI - Megalin deficiency induces critical changes in mouse spinal cord development. AB - Low density lipoprotein receptor-related protein, megalin, is a multifunctional lipoproptein receptor expressed by absorptive epithelia for endocytosis of numerous ligands. Megalin is widely expressed during embryonic life and is essential for development of the nervous system as evidenced by severe forebrain abnormalities in megalin (-/-). Here, we investigated the influence of megalin deficiency on prenatal spinal cord development in mice. In contrast to wild-type mice, cells expressing Olig2 and NG2, that is, oligodendroglial precursor cells, are absent from embryonic stage E16 in megalin (-/-) mice. At the end of prenatal development, there is a failure in vertebral development, and the number of astrocytes are markedly reduced in megalin (-/-) mice. These findings indicate that megalin is essential in astro-oligodendroglial interactions during development of the spinal cord. PMID- 18388739 TI - Are otolithic inputs interpreted better in figure skaters? AB - The aim of this study was to investigate whether figure skaters, as individuals who experience intense vestibular stimulation, presented modification of the otolith-ocular reflex. The reflexes of 12 figure skaters were assessed using off vertical axis rotation (OVAR). Horizontal otolith-ocular reflex during OVAR is characterized by two parameters: the eye velocity horizontal modulation, assumed to compensate for perceived lateral linear translation, and the bias, assumed to compensate for the perceived rotation. We observed that skaters presented smaller amplitude of modulation and truly compensatory bias compared with control participants. Thus, the otolithic signal during OVAR seems to be interpreted more as rotation and less as translation or inclination in figure skaters. PMID- 18388740 TI - Auditory sensory memory in 2-year-old children: an event-related potential study. AB - Auditory sensory memory is assumed to play an important role in cognitive development, but little is known about it in young children. The aim of this study was to estimate the duration of auditory sensory memory in 2-year-old children. We recorded the mismatch negativity in response to tone stimuli presented with different interstimulus intervals. Our findings suggest that in 2 year-old children the memory representation of the standard tone remains in the sensory memory store for at least 1 s but for less than 2 s. Recording the mismatch negativity with stimuli presented at various interstimulus intervals seems to be a useful method for studying the relationship between auditory sensory memory and normal and disturbed cognitive development. PMID- 18388741 TI - Cognitive conflict in audiovisual integration: an event-related potential study. AB - This study used event-related potentials (ERPs) to investigate the electrophysiological correlates of cognitive conflict in audiovisual integration during an audiovisual task. ERP analyses revealed: (i) the anterior N1 and P1 were elicited in both matched and mismatched conditions and (ii) audiovisual mismatched answers elicited a more negative ERP deflection at 490 ms (N490) than matched answers. Dipole analysis of the difference wave (mismatched minus matched) localized the generator of the N490 to the posterior cingulate cortex, which may be involved in the control and modulation of conflict processing of Chinese characters when visual and auditory information is mismatched. PMID- 18388742 TI - Infants' attention is biased by emotional expressions and eye gaze direction. AB - This study investigates infants' processing of emotional expressions in combination with referential eye gaze cues. In experiment 1, 7-month-old infants' neural responses to fearful and neutral faces, which were looking at a novel object, were assessed. Infants' attention, as indexed by the negative central component of the event-related potential, was enhanced when the adult gazed at the object with a fearful expression compared with a neutral expression. In experiment 2, no effect of emotion on amplitude of the negative central was found when the face directed eye gaze at the infant and away from the object. We conclude that by 7 months, infants use emotional expressions in triadic person object-person contexts to detect threat in the environment. PMID- 18388743 TI - Expertise and own-race bias in face processing: an event-related potential study. AB - Humans remember own-race faces more accurately than other-race faces (own-race bias). This effect is reduced by expertise with other-race faces. This study examined event-related potentials to own-race and other-race faces in a group of experts to other-race faces and a nonexpert control group. Both groups exhibited own-race recognition biases. Other-race faces elicited more negative and delayed N170 components, suggesting enhanced configural processing of own-race compared with other-race faces. Moreover, an increased P2 (approximately 210-240 ms) was observed for own-race faces. At right occipitotemporal regions of the experts only this P2 effect was found to be absent. These findings demonstrate an influence of early, presumably presemantic processes on the own-race bias, which are mediated by perceptual expertise with other-race faces. PMID- 18388744 TI - Methadone: does it really have low efficacy at micro-opioid receptors? AB - There is confusion in the literature concerning the relative agonist efficacy of methadone at micro-opioid receptors (MOPrs). Here, we confirm that methadone is a full agonist in guanosine 5'-O-[gamma-thio]triphosphate (GTPgammaS) binding studies. Methadone, however, seems to have low efficacy in studies of MOPr activation of G-protein-gated potassium (GIRK) channels, but this is because it directly inhibits the GIRK channels. Methadone also inhibits alpha2-adrenoceptor activated GIRK channels. Methadone is not a specific GIRK channel blocker. It also inhibits small conductance Ca2+-activated K+ (SK2) channels. We conclude that methadone is a full agonist at MOPrs that, as we and others have shown, induces MOPr desensitization and internalization. PMID- 18388745 TI - Development of chloride homeostasis in albino and pigmented rat visual cortex neurons. AB - Albinism has a profound effect on visual development and visual function. Pharmacologically significant alterations of the two most important chloride transporters--KCC2 (outward transporter) and NKCC1 (inward transporter)- functions were found in albino visual cortex neurons, comprising a higher NKCC1 and a lower KCC2 action. In this study, we compare the early postnatal development of the reversal potential of gamma-aminobutyric acidAR-mediated currents in visual cortex neurons of albino and pigmented rats. At birth we found no differences. At the time of eye opening (second week postnatally) the reversal potential of gamma-aminobutyric acidAR-mediated currents is 15 mV more positive and intracellular Cl- concentration is higher in visual cortex neurons of albinos than of pigmented rats. PMID- 18388746 TI - Tremor varies as a function of the temporal regularity of deep brain stimulation. AB - The frequency of stimulation is one of the primary factors determining the effectiveness of deep brain stimulation (DBS) in relieving tremor. DBS efficacy, however, may depend not only on the average frequency of stimulation, but also on the temporal pattern of stimulation. We conducted intraoperative measurements of the effect of temporally irregular DBS (nonconstant interpulse intervals) on tremor. As the coefficient of variation of irregular high frequency DBS trains increased, they became less effective at reducing tremor (mixed effects regression model, P<0.04). These data provide evidence that the effects of DBS are dependent not only on the average frequency of DBS, but also on the regularity of the temporal spacing of DBS pulses. PMID- 18388747 TI - The train is off the track. PMID- 18388748 TI - Minimally invasive plate osteosynthesis of distal radius fractures using a pronator sparing approach. AB - Volar locking plates for fractures of the distal radius are a common option of care in many centers. Currently, these plates are placed through a standard approach, such as the flexor carpi radialis interval. However, in our opinion, the use of conventional plate application techniques is associated with more soft tissue dissection than is necessary for these new plates. This may contribute to postoperative pain and also to scarring, which may impede the range of motion. To avoid this, dorsal plates have been developed that can be inserted percutaneously. Historically, dorsal plating of the distal radius has been associated with increased morbidity relative to volar plates due to soft tissue depth. This article discusses a single surgeon's technique for insertion of volar locking plates through minimal incisions with sparing of the soft tissues. PMID- 18388749 TI - Augmented (Massachusetts General Hospital) Becker technique combined with static splinting in extensor tendons repairs zones III to VI: functional outcome at three months. AB - PURPOSE: The purpose of this study was to investigate the value of the augmented Massachusetts General Hospital (MGH) Becker extensor tendon repair, a strong 4 strand technique that allows mobilization of repaired tendons after only 3 weeks of postoperative static splinting. METHODS: In the prospective study, we used the augmented Becker (MGH) suturing technique with 4-0 Ethilon. Eighteen patients had extensor tendon lacerations repaired with augmented Becker (MGH) technique. Results were compared with the uninjured hand and statistically evaluated. Postoperatively patients were immobilized 3 weeks in a volar splint and then fully mobilized with physiotherapy. Three months postoperatively, all patients had final assessment for total active motions of fingers; extensor lag; range of motion for the metacarpophalangeal, proximal interphalangeal, and distal interphalangeal joints; and pulp-to-palm distance. RESULTS: Eighteen patients with 20 tendon lacerations were included in the study. At the final assessment, average total active motion of fingers was 268 degrees without a statistically significant difference between this and the uninjured side (P = 0.0584). The average pulp-to-palm distance was 0 cm. There were no wound complications or ruptures of repaired tendons. CONCLUSIONS: Augmented (MGH) Becker technique is an adequate 4-strand extensor tendon repair technique that allows mobilization of patients after only 3 weeks of static splinting postoperatively. Injured fingers can then be safely mobilized with expected return of function at 3 months postoperatively. PMID- 18388750 TI - Surgical technique of pedicled bipolar pectoralis major transfer for reconstruction of elbow flexion in brachial plexus palsy. AB - In Erb (C5-C6) type of palsy due to traumatic brachial plexus injury, function is dependent upon the ability of the arm to maneuver and stabilize the hand for action. Complete loss of power of elbow flexion after brachial plexus injuries is a severe disability. If there is control of the shoulder, elbow flexion is of major importance. Various types of procedures have been described to recover or reanimate elbow flexion for chronic or failed upper type paralysis, including free muscle transfer, unipolar and bipolar pedicled muscle transfer, and various tendon transfers. In this article, we describe the surgical technique of bipolar pectoralis major transfer and our experience with this type of transfer, wherein we prefer to transfer all components of the pectoralis major muscle including the sternocostal and clavicular origins and humeral insertion without a fascial graft by the inclusion of a strip of the rectus abdominis fascia along with the flap. PMID- 18388751 TI - Nerve transfers in the hand and upper extremity surgery. AB - Modern nerve-to-nerve transfers represent one of the greatest advances in peripheral nerve surgery. Lessons of tendon transfers have taught that nerves to specific musculotendinous units are expendable, and greater understanding of peripheral nerve topography has revealed redundant fascicles in peripheral nerves. Transfer of these redundant or expendable nerves to recipient nerves close to the end organ allows for earlier reinnervation and preservation of those musculotendinous units. Such nerve transfers provide significantly better treatment options in many cases of nerve injury where previous outcomes were expected to be poor, such as with proximal injuries, long nerve gaps, or unavailability of the proximal injured segment. This article will review current nerve transfers in the hand and upper extremity. PMID- 18388752 TI - Pronating radius osteotomy for supination deformity in children with obstetric brachial plexus palsy. AB - PURPOSE: In obstetric brachial plexus lesions, muscle imbalance caused by active supinator muscles and paralyzed pronator muscles can result in a supination position of the wrist, which, apart from cosmesis, may interfere with function. METHODS: In this retrospective study, we describe the results of a pronating radius osteotomy for supination deformity of the hand in children with an obstetric brachial plexus lesion. RESULTS: After a mean follow-up of 23 months, all 8 patients (mean age, 9.4 years; range, 4-13 years), operated between 1998 and 2006, had improved functionally and aesthetically. CONCLUSIONS: All patients had improved functionally and aesthetically. PMID- 18388753 TI - Arthroscopic hemitrapeziectomy with tendon interposition for arthritis at the first carpometacarpal joint. AB - The first carpometacarpal (CMC) joint, also referred to as trapeziometacarpal joint, is the area of the hand most commonly symptomatic of osteoarthritis. Although there are a variety of surgical techniques that treat this condition, this article focuses on the technical aspects of arthroscopic hemitrapeziectomy with tendon interposition. Furthermore, this study evaluated the use of arthroscopy to treat CMC arthritis, with the expectation that an arthroscopic procedure would lead to low morbidity, quick recovery of function, rapid resolution of pain, and satisfactory results in patients' strength, range of motion, and pain relief. Early outcomes data indicate that all patients experienced statistically significant improvement in their pain scale rating at a mean of 11 months after the operation. All patients were satisfied with the outcome of their surgery. All patients would choose to have this surgery again. This study supports arthroscopic hemitrapeziectomy with tendon interposition as a safe and effective treatment for CMC arthritis. PMID- 18388754 TI - Minimally invasive intramedullary rod fixation of multiple metacarpal shaft fractures. AB - Metacarpal shaft fractures tend to shorten and angulate. This tendency is accentuated with the fracture of multiple metacarpals. A variety of methods for treatment have been described. The purpose of this study is to present the results of treatment in patients with multiple metacarpal shaft fractures, treated in a minimally invasive manner, with an intramedullary rod device. PMID- 18388755 TI - The reverse shoulder prosthesis--surgical technique. AB - In this article, the surgical technique of the reverse shoulder prosthesis is described in detail. Indications for the procedure, difficulties encountered during surgery, and potential postoperative complications and their solutions are discussed. PMID- 18388756 TI - Fat-covered vein graft. AB - Vein graft is widely used in peripheral vascular surgery. Many free flaps and replantation failures are the result of technical problems in performing the anastomoses. A remarkable number of these technical errors are caused by tension or improper placement of the surgical union. Use of a vein graft may prevent tensioning and give us a more accessible repair site. However, the use of a graft not only doubles the repair side, but may also increase the risk of circulatory disturbance. Many technical, mechanical, and histological factors were cited as factors in the formation of thrombus. The use of a vein graft with a fatty tissue cover may prevent many of the potential risk factors. PMID- 18388757 TI - Treatment of complete acromioclavicular separations with a modified Weaver and Dunn technique. AB - The purpose of this article is to describe the indications, operative technique, and postoperative rehabilitation protocol for treatment of complete acromioclavicular separations. A modified Weaver and Dunn technique is described, with a detailed harvesting procedure of the coracoacromial ligament, along with a wafer of bone for anatomical reconstruction of the coracoclavicular ligaments. A suture anchor is used to provide temporary stability to the acromioclavicular joint while the transferred coracoacromial ligament heals in the new position. PMID- 18388758 TI - Flag dressings for pediatric upper limb surgery. PMID- 18388759 TI - Radioscapholunate arthrodesis for advanced degenerative radiocarpal osteoarthritis. PMID- 18388760 TI - New insights into the comorbidity between ADHD and major depression in adolescent and young adult females. AB - OBJECTIVE: The main aim of this study was to evaluate the association between attention-deficit/hyperactivity disorder (ADHD) and major depression (MD) in adolescent and young adult females. METHOD: Subjects were females with (n = 140) and without (n = 122) ADHD ascertained from pediatric and psychiatric settings. Subjects were followed prospectively for 5 years into adolescence and young adulthood and reassessed in multiple nonoverlapping domains including psychiatric, cognitive, interpersonal, family, and educational functioning. RESULTS: Females with ADHD had a 2.5 times higher risk for MD at adolescent follow-up compared with control females, adjusting for psychiatric comorbidity. MD in females with ADHD was associated with an earlier age at onset, greater than twice the duration, more severe depression-associated impairment, a higher rate of suicidality, and a greater likelihood of requiring psychiatric hospitalization than MD in control girls. Parental MD and proband mania were significant predictors of MD among females with ADHD, independently of other predictors. CONCLUSIONS: MD emerging in the context of ADHD in females is an impairing and severe comorbidity worthy of further clinical and scientific considerations. PMID- 18388761 TI - Depression in men in the postnatal period and later child psychopathology: a population cohort study. AB - OBJECTIVE: Postnatal depression in women is associated with adverse effects on both maternal health and children's development. It is unclear whether depression in men at this time poses comparable risks. The present study set out to assess the association between depression in men in the postnatal period and later psychiatric disorders in their children and to investigate predisposing factors for depression in men following childbirth. METHOD: A population-based cohort of 10,975 fathers and their children from the Avon Longitudinal Study of Parents and Children (ALSPAC) was recruited in the prenatal period and followed for 7 years. Paternal depressive symptoms were assessed with the Edinburgh Postnatal Depression Scale and later child psychiatric disorder (DSM-IV) with the Development and Well-Being Assessment. RESULTS: Depression in fathers in the postnatal period was significantly associated with psychiatric disorder in their children 7 years later (adjusted OR 1.72, 95% CI 1.07-2.77), most notably oppositional defiant/conduct disorders (adjusted OR 1.94, 95% CI 1.04-3.61), after adjusting for maternal depression and paternal educational level. A history of severe depression and high prenatal symptom scores for depression and anxiety were the strongest predictors of paternal depression in the postnatal period. CONCLUSIONS: Depression in fathers in the postnatal period is associated with later psychiatric disorders in their children, independently of maternal postnatal depression. Further research into the risks associated with paternal psychopathology is required because this could represent an important opportunity for public health intervention. PMID- 18388762 TI - Attentional bias for emotional faces in children with generalized anxiety disorder. AB - OBJECTIVE: To examine attentional bias for angry and happy faces in 7- to 12-year old children with generalized anxiety disorder (GAD; n = 23) and nonanxious controls (n = 25). METHOD: Children completed a visual probe task in which pairs of face stimuli were displayed for 500 milliseconds and were replaced by a visual probe in the spatial location of one of the faces. RESULTS: Severely anxious children with GAD showed an attentional bias toward both angry and happy faces. Children with GAD with a milder level of anxiety and nonanxious controls did not show an attentional bias toward emotional faces. Moreover, within the GAD group, attentional bias for angry faces was associated with increased anxiety severity and the presence of social phobia. CONCLUSIONS: Biased attention toward threat as a function of increased severity in pediatric GAD may reflect differing threat appraisal processes or emotion regulation strategies. PMID- 18388763 TI - Father-child transmission of antisocial behavior: the moderating role of father's presence in the home. AB - OBJECTIVE: To demonstrate an environmental effect of being raised by an antisocial father and to test whether the transmission of antisocial behavior from father to child is moderated by the father's presence in the home. METHOD: A community sample of male and female 11- and 17-year-old twins and their biological parents participating in the Minnesota Twin Family Study was used. A series of hierarchical linear regression models was used to examine the relationship between father antisociality and his children's externalizing psychopathology and to determine whether the father's time spent in the home moderated this relationship. Models controlled for the child's sex. RESULTS: A significant main effect of both father's antisociality and father's presence on the children's externalizing psychopathology was found: Children born to antisocial fathers evidenced higher rates of externalizing behavior, and children raised without their biological father in the home exhibited more externalizing behaviors. The interaction was also significant such that the association between father and child antisociality was stronger when the father was present for a longer period of the child's life. Furthermore, when fathers show high levels of antisociality, fathers' presence appears to have deleterious rather than beneficial effects on child behavior. CONCLUSIONS: The present results suggest the transmission of antisociality from father to child is at least partially environmentally moderated. PMID- 18388764 TI - Neuropsychological function in adolescent girls with conduct disorder. AB - OBJECTIVES: Conduct disorder (CD) in adolescent girls is a significant problem, but few data exist on the neuropsychological function of girls with this disorder. We investigated whether girls with CD have poorer neuropsychological function than girls without any psychiatric disorder, whether these differences remained significant after adjusting for demographics, and whether psychiatric comorbidity, age at onset of CD symptoms, or aggressive CD behaviors were differentially associated with neuropsychological function within the CD group. METHOD: Ninety-three girls, ages 15 to 17 years, from the community (52 CD; 41 without any disorder [normal controls]) received a neuropsychological battery examining motor/laterality, general intelligence, language, visuospatial, visual motor, executive function, and academic achievement domains. RESULTS: Girls with CD had lower general intelligence and poorer performance on visuospatial, executive function, and academic achievement domains. After adjusting for demographic factors, scores in the CD group were worse for general intelligence and in the visuospatial and academic achievement domains. Comorbid substance use disorder was negatively correlated with motor/laterality; no other intra-CD group comparisons were statistically significant. CONCLUSIONS: Girls with CD had deficits in several domains of neuropsychological function. Possible explanations for the findings and clinical implications are discussed. PMID- 18388765 TI - Predictors and moderators of outcome in family-based treatment for adolescent bulimia nervosa. AB - OBJECTIVE: To explore the predictors and moderators of treatment outcome for adolescents with bulimia nervosa (BN) who participated in family-based treatment or individual supportive psychotherapy. METHOD: Data derived from a randomized controlled trial (n = 80) of family-based treatment of BN and supportive psychotherapy were used to explore possible predictors and moderators of treatment outcome. RESULTS: Participants with less severe Eating Disorder Examination eating concerns at baseline were more likely to have remitted (abstained from binge eating and purging) after treatment (odds ratio [OR] 0.47; p <.01) and follow-up (OR 0.53; p <.01), regardless of the treatment that they received. Participants with lower baseline depression scores were more likely to have partial remission (no longer meeting study entry criteria) after treatment (OR 0.93; p <.01), whereas those with fewer binge-eating/purging episodes at baseline were more likely to have partial remission at follow-up (OR 0.98; p <.05). In terms of moderators, participants with less severe eating disorder psychopathology (Eating Disorder Examination global score), receiving FBT-BN, were more likely to meet criteria for partial remission at follow-up (OR 0.44; p <.05). CONCLUSIONS: Lower eating concerns are the best predictor of remission for adolescents with BN, and family-based treatment of BN may be most effective in those cases with low levels of eating disorder psychopathology. PMID- 18388766 TI - Change in child psychopathology with improvement in parental depression: a systematic review. AB - OBJECTIVE: To systematically review current research evidence of associations between improvement in parents' depression and their children's psychopathology. METHOD: Relevant studies were identified using PsycINFO (1806-2007) and Medline (1950-2007). The search terms used were "depression," "postpartum depression," "treatment," "mothers," "fathers," "parents," "offspring," "mother child relations," and "father child relations." The reference sections of identified articles were also examined for additional relevant articles. Open and controlled clinical trials and observational studies of depressed parents that also included psychological and behavioral assessments of offspring 18 years of age or younger were included in the review. RESULTS: Ten studies meeting broad criteria for inclusion were reviewed. These studies varied considerably in sample, treatment, assessment, and analysis. Based on the few studies, there is some evidence of associations between successful treatment of parents' depression and improvement in children's symptoms and functioning, but treatment of postpartum depression may not be sufficient for improving cognitive development, attachment, and temperament in infants and toddlers. CONCLUSIONS: Due to the public health implications of the findings, further study of the effects of improvement in parental depression on child psychopathology is warranted. These studies need to examine the precise relation between parental and child symptoms, the differential effect of parents' treatment with psychotherapy versus medication, the effect of fathers' as well as mothers' symptomatic improvement on children, and mediators and moderators of the relation between parental improvement and child psychopathology. PMID- 18388767 TI - Reduced eye gaze explains "fear blindness" in childhood psychopathic traits. AB - OBJECTIVE: Damage to the amygdala produces deficits in the ability to recognize fear due to attentional neglect of other people's eyes. Interestingly, children with high psychopathic traits also show problems recognizing fear; however, the reasons for this are not known. This study tested whether psychopathic traits are associated with reduced attention to the eye region of other people's faces. METHOD: Adolescent males (N = 100; age mean 12.4 years, SD 2.2) were stratified by psychopathic traits and assessed using a Tobii eye tracker to measure primacy, number, and duration of fixations to the eye and mouth regions of emotional faces presented via the UNSW Facial Emotion Task. RESULTS: High psychopathic traits predicted poor fear recognition (1.21 versus 1.35; p < .05) and lower number (1.85 versus 2.51; p < .001) and duration (375 versus 620 ms; p < .001) of eye fixations, and fewer first foci to the eye region (1.01 versus 2.01; p < .001). There were no differences in gaze indices to the mouth region. All indices of gaze to the eye region correlated positively with accurate recognition of fear for the high psychopathy group, especially the number of times that subjects looked at the eyes first (r = .50; p < .01). CONCLUSIONS: Attention to other people's eyes is reduced in young people with high psychopathic traits, thus accounting for their problems with fear recognition, and is consistent with amygdala dysfunction failing to promote attention to emotional salience in the environment. PMID- 18388768 TI - Change in adolescents' internalizing symptomatology as a function of sex and the timing of maternal depressive symptomatology. AB - OBJECTIVE: The purpose of this exploratory study was to examine change in internalizing symptoms from late childhood (age 10) into mid-adolescence (age 15) in a nationally representative sample of Canadian children. The roles of a child's sex, maternal depressive symptoms in late childhood, and their interactions were investigated. METHOD: The sample was derived from the National Longitudinal Survey of Children and Youth. Mothers reported on their own depressive symptoms and children reported on their own internalizing symptoms at three time (T) points (T1: 1994/1995; T2: 1996/1997; T3: 1998/1999). Change in children's internalizing symptoms was investigated using multiple regression. RESULTS: Girls increased and boys decreased in their internalizing symptoms from T1 to T3. The effect of maternal depressive symptoms at T1 was moderated by sex and remained significant after controlling for maternal depressive symptoms at T2 and T3, with more adverse effects in girls. CONCLUSIONS: The internalizing symptoms of girls increased from childhood to adolescence, whereas those for boys decreased. Female children exposed to maternal depressive symptoms T1 continued to show negative effects 4 years later. PMID- 18388769 TI - Is the canine fossa puncture approach really necessary for the severely diseased maxillary sinus during endoscopic sinus surgery? AB - OBJECTIVES: To evaluate the necessity of canine fossa puncture (CFP) by comparing the symptom scores and postoperative computed tomography (CT) findings between patients with severe maxillary sinus disease who underwent CFP and those who underwent maxillary sinus clearance through a middle meatal antrostomy (MMA). STUDY DESIGN: A prospective, randomized study. MATERIALS AND METHODS: Fourteen patients met the inclusion criteria for each of the CFP and MMA groups. In both groups, all diseased sinuses were addressed in the same manner using the same surgical techniques; the only difference was management of the maxillary sinus. Most of the patients completed the Sinonasal Outcome Test 20 (SNOT-20) and visual analogue scales (VAS) for the six main symptoms preoperatively and 3, 6, and 12 months postoperatively. The Lund-Mackay scores for the maxillary sinus and for all sinuses were calculated from the preoperative CT scan and another scan taken 12 months postoperatively. The mucosal thickening as a percentage of the total volume of the maxillary sinus was also evaluated on the postoperative CT scans, and complications related to both procedures were investigated. RESULTS: Twenty four patients completed the follow-up, questionnaires, and postoperative CT scans and were included in the analysis: 11 CFP patients and 13 MMA patients. All of the patients had chronic rhinosinusitis with nasal polyposis (NP). There were no significantdifferences in polyp extent or Lund-Mackay score for the maxillary sinus and for all sinuses on the pre- and postoperative CT scans between the groups. The volume of mucosal thickening also did not differ significantly between the groups on the postoperative CT scans. The SNOT-20 and VAS scores improved significantly 3, 6, and 12 months after the procedure in both groups. However, there were no significant differences between the two groups except for the VAS for postnasal drip 3 months postoperatively, which was better in the CFP group. Six of 11 patients in the CFP group experienced one or more complications after the procedure, although all of the symptoms resolved spontaneously within 3 months. In the MMA group, three patients had bleeding from the branches of the sphenopalatine artery during widening of the ostium, which was controlled intraoperatively with suction cauterization. CONCLUSIONS: We could not find any benefits of the CFP procedure over the conventional MMA method in the present study. Although CFP is a useful method for removing severe mucosal disease that cannot be reached through the MMA, it does not guarantee a better subjective or objective surgical outcome in patients who have accompanying NP. PMID- 18388770 TI - The effect of topical doxycycline in the prevention of experimental tympanosclerosis. AB - OBJECTIVES: The aim of this study is to determine the effectiveness of topical doxycycline used in the process of experimental myringosclerosis and tympanosclerosis. STUDY DESIGN: A prospective experimental animal study. METHODS: Experimental tympanosclerosis was accomplished in 25 healthy adult guinea pigs by inoculation with 2.5 x 10(7) colony-forming units of type-3 Streptococcus pneumoniae microorganisms followed by bilateral myringotomy. While the animals' right ears received a topical doxycycline treatment daily, their left ears were left untreated and used as controls. Otomicroscopic examination was carried out weekly and healing tympanic membranes were remyringotomized. After a 6-week follow-up, the temporal bones of 24 of 25 animals were removed and light microscopy examination was done regarding tympanic membrane myringosclerosis and middle ear mucosal sclerosis. RESULTS: Myringosclerosis was noticed to a lesser extent in the doxycycline-treated group when compared to the untreated control group. Light microscopy evaluation revealed a difference in the area and thickness of the sclerotic plaques of myringosclerosis of the tympanic membranes in the doxycycline-treated group and the control group, being significantly smaller and thinner in the treated group (P < .001, P < .04, respectively). Similarly, the area and thickness of the sclerotic plaques in the middle ear mucosa were significantly smaller and thinner in the doxycycline treated group (P < .001, P < .03). CONCLUSION: This study demonstrated that the potent matrix metallo-proteinase inhibitor doxycycline plays a preventive role in the development of experimentally induced tympanosclerosis. PMID- 18388771 TI - Variations in posttonsillectomy hemorrhage rates are scale invariant. AB - BACKGROUND: Scale invariance is a property of scientific laws or objects that change in a prescribed fashion if measurements are scaled, and is often represented by a power-law relationship. Power laws suggest that events of a large magnitude will be rare, while small events will be much more common, and that a simple mathematical law relates "severity" with frequency. Scale invariance has been demonstrated in scientific fields including physics, social science, and economics. The authors use the complication of a posttonsillectomy hemorrhage to test whether this property is a feature of surgical complications. METHODS: Non-identifiable data were obtained regarding posttonsillectomy hemorrhage and subcategorized by calendar month, and the percentage rate of posttonsillectomy hemorrhage was calculated. The data were then transformed using a logarithmic function. This transformed data were plotted and a linear regression analysis was performed. RESULTS: The 13-year period studied included 6,381 tonsillectomy procedures. The logarithm of the frequency of a given rate range of posttonsillectomy hemorrhage (y) was linearly related to the logarithm of the geometric mean of the rate range (x). The best-fit straight line was y = 1.3996x + 2.0624 with R2 = 0.851, n = 10, r = 0.922, and P < .001. CONCLUSIONS: The authors found that the incidence of posttonsillectomy hemorrhage is scale invariant. The practical implication is that the observation of rare incidences of large hemorrhage rates may not be due to a unique circumstance or a particular operative fault. To reduce the incidence of extreme rates of postoperative hemorrhage, a review of the entire process of tonsillectomy would be required. Scale-invariance analysis may represent a novel tool that should be considered when reviewing surgical complications. PMID- 18388772 TI - Developing a rabbit model of rhinogenic chronic rhinosinusitis. AB - OBJECTIVE: The purpose of this study was to develop a rabbit model of rhinogenic chronic rhinosinusitis (CRS). METHODS: New Zealand white rabbits were used and divided into two groups. In group A rabbits, a piece of Merocel (Medtronic-Xomed, Jacksonville, FL) was inserted into one nasal cavity and the other was left undisturbed as control. In group B rabbits, 1 microg phorbol 12-myristate 13 acetate (PMA) was injected into bilateral nasal lateral walls and then a piece of Merocel (Medtronic-Xomed) was inserted into one nasal cavity. At week 2, the Merocel (Medtronic-Xomed) was removed, and computed tomography (CT), nasoendoscopy, and cultures were performed. All examinations were repeated at week 12. Rabbits that had purulent discharge in nasal cavities and sinuses opacification shown in CT scans were diagnosed as having rhinosinusitis. Rabbits with CRS were randomly allocated to receive intravenous ceftriaxone (50 mg/kg/day) for 28 days or nothing. All rabbits with CRS received CT scans, nasoendoscopy, and cultures at week 16. RESULTS: At week 12, CRS had developed in two controlled nasal cavities, six nasal cavities inserted with Merocel (Medtronic-Xomed), six nasal cavities injected with phorbol 12-myristate 13 acetate (PMA), and seven both PMA-injected and Merocel- (Medtronic-Xomed) inserted nasal cavities. Seven of nine treated CRS sides were clear of opacification after treatment. All non-treated CRS sides had persistent diseases at week 16. There was a significant difference in the CRS incidence (P = .00043) and culture rates (P = .027) between treated and non-treated CRS nasal cavities. CONCLUSIONS: Our study developed a rabbit model of rhinogenic CRS. This model is easily performed and is reversible by treatment. PMID- 18388773 TI - Staging and survival analysis for nonsquamous cell carcinomas of the larynx. AB - OBJECTIVE: Determine staging and comparative survival for nonsquamous cell carcinoma (SCC) of the larynx. STUDY DESIGN: Cross-sectional population analysis. METHODS: : Cases of non-SCC supraglottic, glottic, and subglottic cancers were extracted from the Surveillance, Epidemiology and End Results database (1988 2003) and the staging distribution computed. For each site, Kaplan-Meier survivals were compared according to histology, T-stage, and N-stage. Survival for non-SCC cancers for each site were compared among histologies, T-stage, and N stage as well as with age/sex/stage-matched SCC cancers to determine differences in survival. RESULTS: One hundred forty supraglottic non-SCC cases were identified (67% male, mean age 62.0 yr) consisting of 25 neuroendocrine, 25 small cell, and 22 lymphomas (among others), with mean survivals of 44.8, 52.1, and 80.5 months, respectively. Survival was not significantly different according to histology (log-rank, P = .440) but was significantly different according to T stage (P = .029) and N-stage (P < .001). Seventy-eight glottic non-SCC cases were identified (83% male, mean age 66.1 yr) consisting of 32 spindle cell, 13 sarcoma, and 10 small cell (among others), with mean survivals of 114.6, 89.8, and 39.2 months, respectively. Survival was not significantly different according to histology (P = .056) but was significantly different according to T-stage (P = .002) and N-stage (P = .022). Small numbers (26) precluded subglottic analysis. For both supraglottic and glottic sites including all histologies, survival was similar for matched non-SCC and SCC cases (P = .510 and P = .930, respectively). CONCLUSIONS: Non-SCCs are more common in the supraglottic larynx. T-stage and N stage influence survival more than histology alone for these cancers. PMID- 18388774 TI - Cartilage graft laryngotracheoplasty for anterior subglottic stenoses. PMID- 18388775 TI - Characterizing the oculoauriculofrontonasal syndrome. AB - Human dysmorphology syndromes are frequently defined by characteristic abnormalities in facial morphogenesis. Two such well recognized syndromes are the oculoauriculovertebral spectrum (OAVS) and frontonasal dysplasia (FND). OAVS is diagnosed on the basis of the presence of typical facial features which can include microtia, preauricular tags, hemifacial microsomia, lateral face clefting, epibulbar dermoids, and upper palpebral colobomata. FND is characterized by ocular hypertelorism, nasal clefting, and anterior cranium bifidum occultum. After the first patient was described with features of both OAVS and FND, at least a further 25 patients presenting the 'oculoauriculofrontonasal syndrome' (OAFNS) have been reported. We report on four more patients with OAFNS and review their features, together with those of the other patients reported in the medical literature. We suggest that, statistically, OAFNS is more likely to be a sporadically occurring condition rather than an inherited autosomal recessive trait, as previously suggested. We cannot, however, definitively exclude the possibility of autosomal dominant transmission. Considering the question of whether OAFNS is a part of OAVS, FND, or a distinct clinical entity, we conclude that, for the time being, OAFNS should be considered to be a distinct syndrome, to further our understanding of the aetiology of these conditions. PMID- 18388776 TI - Ocular colobomata, polydactyly, cleft palate and panhypopituitarism: a new syndrome. AB - We describe a male patient with ocular colobomata, cleft palate, polydactyly, panhypopituitarism and possible craniosynostosis, whom we have followed for 30 years. Although there are some similarities to other documented syndromes, this previously unreported combination of features appears to constitute a new syndrome. PMID- 18388777 TI - Familial multiple ventricular extrasystoles, short stature, craniofacial abnormalities and digital hypoplasia: a further case of Stoll syndrome? AB - We report two brothers, their mother and a maternal cousin who had a distinctive facial phenotype, mild brachydactyly and prominence of the interphalangeal joints. One brother and the mother also had multiple ventricular extrasystoles. Six other relatives in four generations were probably affected on the basis of history and family photographs. We also report a further individual from a different family with a similar facial phenotype, Pierre-Robin sequence, tapering fingers and multiple ventricular extrasystoles. These families have some similarities to those reported by Stoll et al. in a single family, showing dominant inheritance. Our patients would seem to have the same or a related condition. PMID- 18388778 TI - Dyssegmental dysplasia in a South African neonate. AB - A female neonate born to nonconsanguineous Zulu parents had dyssegmental dysplasia, Silverman-Handmaker type. This condition has not previously been reported from the continent of Africa. She died at the age of 4 months following the development of pneumonia complicated by an unexplained anaemia. PMID- 18388779 TI - De Barsy syndrome: a review of the phenotype. AB - De Barsy syndrome is a rare, autosomal recessive syndrome characterised by a progeria-like appearance with distinctive facial features and cutis laxa. Ophthalmological, orthopaedic and neurological abnormalities are also typically present. The syndrome was first described by de Barsy et al. in 1967 and since that time approximately 27 further cases have been reported worldwide. We present a case that demonstrates the typical clinical and histological features of de Barsy syndrome. A female infant, the second child of first-cousin parents from a multiply consanguineous family of Pakistani origin, presented at birth with growth retardation, cutis laxa and a progeria-like appearance. She had thin, overlapping fingers and adducted thumbs, blue sclerae, cloudy corneas and myopia. She has failed to thrive and has marked developmental delay and abnormal athetoid movements. During the first year of life she developed pectus excavatum and her facial appearance became more aged. To our knowledge there are no previous reports of de Barsy syndrome in individuals of Pakistani origin. PMID- 18388780 TI - Congenital swan neck deformity of fingers with syndactyly. AB - Congenital swan neck deformity of the fingers is an uncommon congenital disorder. It is sporadic and not associated with other malformations. We report a case of congenital swan neck deformity of the fingers in a 16-year-old boy, with associated ulnar deviation of the fingers, bilateral simian creases and soft tissue syndactyly. PMID- 18388781 TI - The fifth female patient with Myhre syndrome: further delineation. AB - We report the fifth female patient with Myhre syndrome (MS) and review the literature. She is a 13-year-old Mexican girl whose phenotype fulfills all the clinical and radiological criteria reported in MS such as typical facies, short stature, limited joint mobility, and short hands and feet. The physical habitus of MS is described and a square body shape is clearly distinguished in all cases. As the typical muscular build is present mainly in male patients, the MS muscular appearance may be hormonally influenced. PMID- 18388782 TI - The Walker-Warburg phenotype and genotype among Irish Travellers. PMID- 18388783 TI - Facial asymmetry associated with small and large intestinal atresia, and ipsilateral malformations of eye, skin, and extremities. PMID- 18388784 TI - Neuronal migration disorders, agenesis of corpus callosum, preauricular skin tag and bilateral morning glory syndrome in a term newborn infant. PMID- 18388785 TI - Functional pulmonary atresia in a patient with neonatal Marfan syndrome caused by a c.3602G>A mutation in exon 29 of the FBN1 gene. AB - Neonatal Marfan syndrome is a severe form of the syndrome mostly caused by de novo mutations in the fibrillin-1 gene. We report a newborn with neonatal Marfan syndrome and functional pulmonary atresia who died from congestive heart failure on postnatal day 22 despite treatment. He had a mutation in exon 29 of the fibrillin-1 gene at position c.3602G>A. Functional pulmonary atresia may be a life-threatening cardiovascular manifestation of neonatal Marfan syndrome. PMID- 18388786 TI - Caudal appendage in focal dermal hypoplasia (Goltz syndrome). PMID- 18388787 TI - Prune belly syndrome: expanding the phenotype. PMID- 18388788 TI - Persistent cloaca associated with a duplicated left leg: a novel disorganization like syndrome. PMID- 18388789 TI - Goldenhar syndrome with duodenal atresia: a new finding. PMID- 18388790 TI - Congenital microgastria in association with Pierre-Robin sequence. PMID- 18388791 TI - Mandibulofacial dysostosis, acral anomalies and frontonasal dysplasia: a new form of acrofacial dysostosis. AB - We describe a stillborn female with acrofacial dysostosis and frontonasal dysplasia. She had protrusion of the forehead, with marked hypertelorism and absence of the nose but with the rhinencephalon present. Autopsy showed wide cranial sutures, severe hydrocephalus with separation of the right and left hemispheres of the brain, preservation of the olfactory bulb and first and second cranial nerves. The child also had small kidneys bilaterally, rectal atresia and an absent anus with rectovaginal fistula. These clinical findings suggest a new form of acrofacial dysostosis. PMID- 18388792 TI - Oral teratoma, dextrocardia, and congenital heart defect: a nonrandom association or serendipity? PMID- 18388793 TI - Spondylocostal dysostosis associated with diaphragmatic hernia and neural tube defects. PMID- 18388794 TI - Septo-optic dysplasia with a transverse limb defect. PMID- 18388796 TI - B-cell-targeted treatment for multiple sclerosis: mechanism of action and clinical data. AB - Strategies for treating autoimmune disorders are increasingly employing targeted therapies rather than non-specific, multitargeted treatments. Accumulating evidence on the involvement of B lymphocytes in the pathophysiology of autoimmune demyelinating disease has led to a renewed interest in B cells as potential therapeutic targets. In particular, antigen presentation between B cells and T cells, increased trafficking of B cells across the blood-brain barrier, and autoantibodies produced by plasma cells may contribute to the pathophysiology of autoimmune disorders such as multiple sclerosis. Several B-cell-targeted, depletion therapies are currently in development, including rituximab, epratuzumab, diphtheria toxin-single chain Fv (DC2219), belimumab, atacicept, abatacept, and abetimus sodium. Of these agents, only rituximab and abatacept have been evaluated in multiple sclerosis patients. Preliminary results of a phase II trial of rituximab in multiple sclerosis suggest that rituximab is well tolerated and significantly reduces the number of gadolinium enhancing lesions over 24 weeks of treatment. Results of an exploratory analysis suggest the potential promise of abatacept 10 mg/kg for multiple sclerosis. It is expected that future clinical trials will establish a role for B-cell-targeted therapies in the treatment of multiple sclerosis and other autoimmune neurological diseases. This article describes the mechanism of action behind B-cell-targeted depletion therapies in development and reviews available clinical data. PMID- 18388800 TI - The natural history of B cells. AB - Multiple sclerosis is a common neurological disorder that represents a significant source of disability. B cells have recently emerged as a novel therapeutic target for multiple sclerosis. The natural development of B cells is characterized by an antigen-independent phase that occurs in the bone marrow and an antigen-dependent phase that takes place in the peripheral lymphoid tissue. The stage of B-cell development can be identified by the presence of specific cell surface markers. Checkpoints are in place to prevent self-reactive B cells from further development and activation. Some self-reactive B cells are able to escape these checkpoints, resulting in a loss of tolerance. B cells may contribute to systemic autoimmunity and the development of autoimmune disease via cytokine production, antigen presentation, and complement activation. In addition, B cells may trigger autoimmune disease via molecular mimicry, which occurs when a single B-cell receptor recognizes both a non-self antigen molecule and a self-molecule. Accumulating data suggest that ectopic proliferation of B cells in the central nervous system may also play a role. Further research is needed to elucidate the pathology of B cells and their role in central nervous system autoimmune diseases, including multiple sclerosis. PMID- 18388801 TI - The role of B cells in multiple sclerosis: rationale for B-cell-targeted therapies. AB - Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system with no clear etiology. Until recently, most studies have emphasized the role of T cells in the pathogenesis of multiple sclerosis. Data suggesting that B cells play a role in the pathogenesis of multiple sclerosis have been accumulating for the past five decades, demonstrating that the cerebrospinal fluid and central nervous system tissues of multiple sclerosis patients contain B cells, plasma cells, antibodies, and immunoglobulins. Data suggest that B cells are involved in antigen capture and presentation to T cells, cytokine production, antibody secretion, demyelination, tissue damage, and remyelination in multiple sclerosis. These advances in the understanding of B cell and antibody roles in the pathophysiology of multiple sclerosis provide a strong rationale for B-cell-targeted therapies. PMID- 18388803 TI - Hypertensive emergencies in pregnancies in underresourced countries. PMID- 18388804 TI - Editorial summary of symposium on hypertensive disorders of pregnancy. AB - PURPOSE OF REVIEW: Hypertensive disorders of pregnancy, particularly the preeclampsia/eclampsia syndrome, remain the leading causes of worldwide pregnancy related maternal and neonatal mortality and morbidity. This group of conditions are a 'riddle wrapped in a mystery inside an enigma' to quote Winston Churchill. We are fortunate to have contributions from leading clinical experts who have devoted many years of their professional careers attempting to solve this conundrum. RECENT FINDINGS: Dr Jack Moodley has provided us with a perspective on clinical management in underresourced countries. Referral to experts, aggressive treatment of hypertension and use of magnesium sulfate improves care. Dr Shennan focuses on the assessment of risk, close antenatal surveillance and timely delivery. Dr Uzan continues to champion the use of aspirin for prevention of preeclampsia, even though the evidence is contradictory. Dr Sibai addresses the lack of evidence for calcium, vitamin C and E in prevention of preeclampsia. Dr Von Dadelszen is developing a new paradigm for the classification of these disorders and emphasizes the importance of evidence-based intervention. SUMMARY: Evidence suggests that treatment of severe hypertension, seizure prophylaxis with magnesium sulfate, and management by experienced healthcare professionals will improve maternal, fetal and neonatal outcomes. Well designed studies will lead to evidence-based improvement in caring for mothers and babies worldwide. PMID- 18388805 TI - Hypertensive disorders of pregnancy: a UK-based perspective. PMID- 18388806 TI - Hypertensive disorders of pregnancy: the United States perspective. PMID- 18388807 TI - Hypertensive disorders of pregnancy: future perspectives. A French point of view. PMID- 18388808 TI - What matters in preeclampsia are the associated adverse outcomes: the view from Canada. PMID- 18388809 TI - Patient choice cesarean delivery: ethical issues. AB - PURPOSE OF REVIEW: We have recently identified three salient questions within the patient choice cesarean delivery controversy. First, is performing cesarean delivery on maternal request consistent with good professional medial practice? Second, how should physicians respond to or counsel patients who request patient choice cesarean delivery? Third, should patient choice cesarean delivery be routinely offered to all pregnant women? RECENT FINDINGS: In a well informed patient, performing a cesarean delivery on maternal request is medically and ethically acceptable. Physicians, as patient advocates and promoters of overall health and welfare of their patients, however, should, in the absence of an accepted medical indication, recommend against medically unindicated cesarean delivery. While we believe that current evidence supports a physician's decision to accede to an informed patient's request for such a delivery, it does not follow that obstetricians should routinely offer elective cesareans to all patients. SUMMARY: When a patient makes a request for an elective cesarean delivery, obstetricians, in their capacity as patient advocate, must help guide their patient through the labyrinth of detailed medical information toward a decision that respects both the patient's autonomy and the physician's obligation to optimize the health of both the mother and the newborn. PMID- 18388810 TI - Perinatal infections and fetal/neonatal brain injury. AB - PURPOSE OF REVIEW: Fetal and newborn brain injury causes lifelong morbidity for the survivors, with high emotional costs to the individual and the family plus a heavy economic burden for society. This is timely and relevant. Techniques are now available to prevent, detect, and treat those central nervous injuries that result from infection. If instituted, these would have beneficial results for both newborn survival and morbidity. RECENT FINDINGS: The new developments and prevention of adverse impacts of maternal infections due to Rubella, Group B Streptococcus, cytomegalovirus, toxoplasmosis, and chorioamnionitis are discussed. SUMMARY: Rubella immunization has been a success in the USA with virtual elimination of wild virus infection. Broader screening policies need to be instituted to lower the morbidities associated with cytomegalovirus and toxoplasmosis. Alternatives to maternal antibiotic prophylaxis in newborn Group B Streptococcus infection need to be evaluated and implemented when proven effective. The most beneficial treatment protocol for chorioamnionitis needs to be determined by prospective study and then instituted. To date, the combination of maternal antibiotics and steroids appears most promising. PMID- 18388811 TI - Fetal growth restriction: a modern approach. AB - PURPOSE OF REVIEW: Fetal growth restriction is a complicated perinatal condition, with multiple causes. It shares common pathophysiologies with other important disorders, such as preeclampsia and abruption. As a group, these conditions associated with ischemic placental disease are responsible for a large percentage of indicated preterm births. The ability to accurately predict, diagnose and manage these pregnancies has significant and far-reaching implications, including potential effects on long-term adult health. RECENT FINDINGS: Placental ischemia is the most common cause of fetal growth restriction. Alterations in placental development are being linked to various angiogenic mediators, which may be of future use in early risk-determination. Until then, the use of ultrasound to accurately diagnose fetal growth restriction and time delivery is the mainstay of management. Research in this area has revealed some commonalities in the deterioration of the growth restricted fetus, but has also indicated that not every affected fetus will follow the same progression in Doppler and other wellbeing parameters. Most importantly, gestational age at delivery is consistently being documented as a critical factor in perinatal morbidity and mortality. SUMMARY: Fetal growth restriction is a late manifestation of early abnormal placental development. Once abnormal Doppler velocimetry is present, surveillance and timing of delivery should be based on the antepartum test results and on the gestational age. PMID- 18388812 TI - Prenatal origins of adult disease. AB - PURPOSE OF REVIEW: Human epidemiological and animal studies show that many chronic adult conditions have their antecedents in compromised fetal and early postnatal development. Developmental programming is defined as the response by the developing mammalian organism to a specific challenge during a critical time window that alters the trajectory of development with resulting persistent effects on phenotype. Mammals pass more biological milestones before birth than any other time in their lives. Each individual's phenotype is influenced by the developmental environment as much as their genes. A better understanding is required of gene-environment interactions leading to adult disease. RECENT FINDINGS: During development, there are critical periods of vulnerability to suboptimal conditions when programming may permanently modify disease susceptibility. Programming involves structural changes in important organs; altered cell number, imbalance in distribution of different cell types within the organ, and altered blood supply or receptor numbers. Compensatory efforts by the fetus may carry a price. Effects of programming may pass across generations by mechanisms that do not necessarily involve structural gene changes. Programming often has different effects in males and females. SUMMARY: Developmental programming shows that epigenetic factors play major roles in development of phenotype and predisposition to disease in later life. PMID- 18388813 TI - Screening for fetal abnormalities with ultrasound. AB - PURPOSE OF REVIEW: This review serves to explore literature published during the past year that has focussed on sonographic screening for fetal abnormalities. RECENT FINDINGS: The evolving nature of screening for fetal aneuploidy has continued to feature strongly in recent literature. There is growing interest in early detailed ultrasound of fetal anatomy at the time of nuchal translucency thickness assessment. Demonstration of tricuspid regurgitation and ductus venosus assessment have been shown to be beneficial additional second-line tests for aneuploidy screening in high-risk patients. Frontomaxillary facial angle assessment has been explored as a new marker for aneuploidy. The limitations of first-trimester nasal bone measurement were reiterated while its measurement has been shown to be beneficial in the second trimester, especially when calculated with multiples of the median. As regards second-trimester screening, studies have explored the effectiveness of congenital heart-defect screening and the efficiency of the genetic sonogram for trisomy 21. The role of nuchal translucency discordance in monochorionic twins has also been readdressed. SUMMARY: Screening for fetal abnormalities continues to evolve with the introduction of novel techniques and the further refinement of previously proposed screening tools. How these modalities are implemented into routine clinical practice remains to be seen. PMID- 18388814 TI - Noninvasive prenatal diagnosis: current practice and future perspectives. AB - PURPOSE OF REVIEW: To review recent developments in the noninvasive determination of fetal genetic loci via the use of fetal cells or cell-free nucleic acids in maternal blood, and provide an overview of the possibilities for future clinical applications. RECENT FINDINGS: Noninvasive prenatal fetal sex or Rhesus D status determination via the analysis of cell-free fetal DNA is now offered by a number of European centres. The detection of fetal loci not completely disparate from maternal ones, such as point mutations, however, is more complex due to the preponderance of maternal cell-free DNA. Promising approaches to overcome this drawback include specialized PCR protocols employing peptide-nucleic acid clamps, mass spectrometry or the enrichment of fetal cell-free DNA sequences by size fractionation. The discovery of cell-free fetal mRNA has opened up the exciting possibility of noninvasive detection of Down's syndrome, as well as examination of fetal gene expression profiles. SUMMARY: The noninvasive determination of Mendelian disorders such as thalassemia or cystic fibrosis, and Down's syndrome may soon become a clinical reality. Noninvasive fetal profiling technologies could lead to the development of a new generation of highly specific tools for the detection of pregnancies at risk for preeclampsia or preterm labour. PMID- 18388815 TI - Fetal health assessment using prenatal diagnostic techniques. AB - PURPOSE OF REVIEW: The assessment of fetal health has occupied our professional attention for many years. As the development of technologies for prenatal diagnostic techniques has progressed, applications of such technologies have aided in the overall assessment of fetal well-being. RECENT FINDINGS: Fetal heart rate monitoring remains the main form of fetal assessment for high-risk pregnancies. The additional assessments afforded by the analysis of ST and T-wave changes of the fetal electrocardiogram hold promise for improving the predictive value of fetal heart-rate assessments. Ultrasound has been invaluable for assessment of fetal anatomy, and the use of Doppler ultrasound has given insight into fetal cardiovascular responses to such conditions as intrauterine growth restriction and fetal anemia caused by red blood cell alloimmunization. SUMMARY: As prenatal diagnostic techniques evolve, their application to the assessment of fetal well-being offers insight into pathophysiologic mechanisms of fetal disease, innovative tools to assess fetal health, and fertile ground for future investigation. PMID- 18388816 TI - Genetic screening and counseling. AB - PURPOSE OF REVIEW: Recent advances in genetic technology have substantial implications for prenatal screening and diagnostic testing. The past year has also seen important changes in recommendations surrounding the genetic counseling that occurs in the provision of such testing. RECENT FINDINGS: Multiple screening tests for single gene disorders, chromosomal abnormalities, and structural birth defects are now routinely offered to all pregnant women. Ethnicity-based screening for single gene disorders includes Tay Sachs disease, cystic fibrosis, and hemoglobinopathies. Recent discussions have involved, not only additional disorders that warrant screening, but a re-evaluation of the paradigm of selecting disorders for population-based screening. Testing for chromosomal abnormalities has seen the introduction of first-trimester screening, as well as strategies to improve detection through sequential testing. Changes in recommendations for screening compared with diagnostic testing, and a move away from maternal age-based dichotomizing of testing, have had major implications for provision of genetic counseling by providers of prenatal care. SUMMARY: Advances in genetic testing have resulted in tremendous benefits to patients, and challenges to providers. New approaches to education and counseling are needed to assure that all patients receive a complete and balanced review of their prenatal genetic-testing options. PMID- 18388817 TI - Chorionic villus sampling and amniocentesis in 2008. AB - PURPOSE OF REVIEW: Over the past decades there have been wide discrepancies between quoted risks of diagnostic procedures (chorionic villus sampling and amniocentesis) yet little properly controlled and randomized data to back often dogmatic assertions. Here, we review the historical and current literature to determine realistic estimates. RECENT FINDINGS: Several papers this past year have addressed in cohort studies and meta-analyses composite risks for both chorionic villus sampling and amniocentesis. The studies have had varying degrees of reliability and likely reproducibility. SUMMARY: Despite one outlier paper, which had major methodological flaws, the consensus of the modern literature is that in experienced hands there is little to no differences between the procedure risks of amniocentesis and chorionic villus sampling. The latter, however, is clearly harder to learn and has a steeper learning curve. PMID- 18388818 TI - Operative fetoscopy in complicated monochorionic twins: current status and future direction. AB - PURPOSE OF REVIEW: The aim of this article is to review the prenatal diagnosis and treatment of complicated monochorionic twins, with emphasis on surgical management via operative fetoscopy. RECENT FINDINGS: Ultrasound is effective in the diagnosis and surveillance of several conditions that complicate monochorionic twins, such as twin-twin transfusion syndrome, twin reversed arterial perfusion sequence, selective intrauterine growth restriction, and discordant anomalous twins. Ultrasound is also useful in identifying risk factors for poor perinatal outcomes and identifying those pregnancies that may benefit from fetal intervention. Operative fetoscopy has emerged as a useful tool in the surgical management of complicated monochorionic twins. Recent findings support the use of laser therapy for the treatment of mid-trimester twin-twin transfusion syndrome at experienced centers. The reduction in hemodynamic stress associated with definitive treatment of twin-twin transfusion syndrome by laser surgery translates to improved short and long-term perinatal outcomes. Pregnancies complicated by twin reversed arterial perfusion sequence that meet surgical criteria benefit from umbilical cord occlusion. A prospective randomized trial regarding the role of laser therapy for selective intrauterine growth restriction is ongoing. SUMMARY: Operative fetoscopy has an important role in the management of complicated monochorionic twins. PMID- 18388819 TI - Timing of injury in the fetus and neonate. AB - PURPOSE OF REVIEW: The aim of this article is to assess evidence for the timing of brain injury associated with clinical encephalopathy in the fetus and newborn. RECENT FINDINGS: Older studies found strong epidemiological links between antenatal factors, including exposure to infection, and acute encephalopathy and later cerebral palsy. In contrast, recent cohort studies using early magnetic resonance imaging and spectroscopy, near-infrared spectroscopy and electroencephalogram monitoring suggest that the majority of brain injury in infants with evidence of encephalopathy occurs in the immediate perinatal period and conversely that it is relatively uncommon for chronic, antepartum injury to present with symptoms at birth. A key experimental study showed that mild infection can lead to long-lasting sensitization of the brain to subsequent mild hypoxia-ischemia. This provides a plausible mechanistic link between some antenatal complications and acute perinatal injury. Consistent with this, randomized controlled trials have demonstrated that therapeutic hypothermia can significantly improve outcome of neonatal encephalopathy at term. Electroencephalogram monitoring seems to be the most promising modality to identify infants who may benefit from potential neuroprotective treatments, but more research is needed to refine its use. SUMMARY: There is now strong evidence that brain injury commonly occurs in the immediate perinatal period, and so may be potentially treatable. PMID- 18388820 TI - Transcervical sterilization. PMID- 18388822 TI - Bibliography. Current world literature. Model systems. PMID- 18388823 TI - Retail-based clinics a viable resource for primary care. PMID- 18388824 TI - Update on the DNP degree. PMID- 18388825 TI - New asthma guidelines encourage more activity and a better night's sleep. PMID- 18388826 TI - Hallucinations during lamotrigine treatment. PMID- 18388829 TI - Pain in the older adult. PMID- 18388831 TI - Community-associated MRSA: coming to a patient near you? PMID- 18388833 TI - Acute abdominal pain: a diagnostic challenge. PMID- 18388834 TI - Improving the quality of nurse practitioner education: the case of Botswana. PMID- 18388837 TI - Access to clinically-detailed patient information: a fundamental element for improving the efficiency and quality of healthcare. PMID- 18388836 TI - Risk-adjusting hospital inpatient mortality using automated inpatient, outpatient, and laboratory databases. AB - OBJECTIVES: To develop a risk-adjustment methodology that maximizes the use of automated physiology and diagnosis data from the time period preceding hospitalization. DESIGN: : Retrospective cohort study using split-validation and logistic regression. SETTING: Seventeen hospitals in a large integrated health care delivery system. SUBJECTS: Patients (n = 259,699) hospitalized between January 2002 and June 2005. MAIN OUTCOME MEASURES: Inpatient and 30-day mortality. RESULTS: Inpatient mortality was 3.50%; 30-day mortality was 4.06%. We tested logistic regression models in a randomly chosen derivation dataset consisting of 50% of the records and applied their coefficients to the validation dataset. The final model included sex, age, admission type, admission diagnosis, a Laboratory-based Acute Physiology Score (LAPS), and a COmorbidity Point Score (COPS). The LAPS integrates information from 14 laboratory tests obtained in the 24 hours preceding hospitalization into a single continuous variable. Using Diagnostic Cost Groups software, we categorized patients as having up to 40 different comorbidities based on outpatient and inpatient data from the 12 months preceding hospitalization. The COPS integrates information regarding these 41 comorbidities into a single continuous variable. Our best model for inpatient mortality had a c statistic of 0.88 in the validation dataset, whereas the c statistic for 30-day mortality was 0.86; both models had excellent calibration. Physiologic data accounted for a substantial proportion of the model's predictive ability. CONCLUSION: Efforts to support improvement of hospital outcomes can take advantage of risk-adjustment methods based on automated physiology and diagnosis data that are not confounded by information obtained after hospital admission. PMID- 18388838 TI - Provider-level effects on psychiatric inpatient length of stay for youth with mental health and substance abuse disorders. AB - OBJECTIVE: Previous research on inpatient care for children and adolescents with emotional or behavioral problems indicates that patient-level factors predict length of stay (LOS) poorly. This analysis examines whether patient-level factors are poor predictors of LOS, because LOS is primarily determined by facilities rather patients. STUDY DESIGN: This study uses Tennessee Medicaid claims data from 1996 to 2001. The data include information on 14,162 observations related to 8400 patients (age 12-21) from 163 hospitals. We estimate log LOS using a cross classified model. Covariates include admission-level characteristics (age, diagnosis, qualification for Medicaid, year), patient-level characteristics (gender, race), and facility characteristics (facility type). PRINCIPLE FINDINGS: Our results suggest that variation in LOS is attributable to facility-level factors (51%), time-invariance patient-level factors (5%), factors that vary across admissions (42%), and a correlation between patient-level and facility level factors (5%). CONCLUSIONS: About half of the variation in LOS is explained by facility-level factors. Given the vulnerable nature of youth who are in need of inpatient psychiatric care, it may be particularly important to monitor provider-level processes and outcomes. Measuring facility or provider level quality is complicated because of difficulties in adjusting for case-mix severity across providers. The methodology presented here represents a general framework that can be widely used in health services research. Potential applications include broadening models of utilization to simultaneously include patient, provider, geographic and community level variations, as well as provider profiling. PMID- 18388839 TI - Evaluation of a patient activation and empowerment intervention in mental health care. AB - BACKGROUND: Evidence suggests that minority populations have lower levels of attendance and retention in mental health care than non-Latino whites. Patient activation and empowerment interventions may be effective in increasing minority patients' attendance and retention. OBJECTIVES: This study developed and evaluated a patient self-reported activation and empowerment strategy in mental health care. RESEARCH DESIGN: The Right Question Project-Mental Health (RQP-MH) trainings consisted of 3 individual sessions using a pre/post test comparison group design with patients from 2 community mental health clinics. The RQP-MH intervention taught participants to identify questions that would help them consider their role, process and reasons behind a decision; and empowerment strategies to better manage their care. SUBJECTS: A total of 231 participated, completing at least the pretest interview (n = 141 intervention site, 90 comparison site). MEASURES: Four main outcomes were linked to the intervention: changes in self-reported patient activation; changes in self-reported patient empowerment; treatment attendance; and retention in treatment. RESULTS: Findings show that intervention participants were over twice as likely to be retained in treatment and over 3 times more likely than comparison participants to have scheduled at least 1 visit during the 6-month follow-up period. Similarly, intervention participants demonstrated 29% more attendance to scheduled visits than comparison patients. There was no evidence of an effect on self-reported patient empowerment, only on self-reported patient activation. CONCLUSIONS: Results demonstrate the intervention's potential to increase self-reported patient activation, retention, and attendance in mental health care for minority populations. By facilitating patient-provider communication, the RQP-MH intervention may help minorities effectively participate in mental health care. PMID- 18388840 TI - The influence of depressive symptoms on clinician-patient communication among patients with type 2 diabetes. AB - BACKGROUND: Depression is associated with poor chronic illness outcomes, but it is unknown whether depression influences the quality of communication during the clinical encounter. We investigated whether diabetes patients with depressive symptoms, compared with those without depressive symptoms, report worse clinician patient communication, and which domains of communication are most affected. METHODS: We surveyed 231 ethnically diverse, English-speaking patients with diabetes to ascertain their experiences of communication with their primary care clinician. We selected measures from the interpersonal processes of care (IPC) instrument to assess communication and dichotomized the 7 subscales into "optimal" or "suboptimal" communication. We used the Clinical Epidemiologic Services for Depression (CES-D 10) to categorize patients as having no (CES-D 10 score <10), mild (CES-D 10 score 10-14), or severe (CES-D 10 score >14) depressive symptoms. We used multivariable logistic regression to evaluate the relationship between depressive symptoms and communication subscales. RESULTS: Thirty-five percent of subjects reported severe depressive symptoms. Compared with those with no depressive symptoms, the presence of severe depressive symptoms was independently associated with suboptimal communication in 4 of 7 subscales: elicitation of patient problems, concerns, and expectations (adjusted odds ratio [AOR], 2.94; 95% confidence interval [CI], 1.14-7.61); explanations of condition (AOR, 3.79; 95% CI, 1.41-10.21); empowerment (AOR, 2.98; 95% CI, 1.35 6.58); and decision-making (AOR, 2.56; 95% CI, 1.14-5.78). CONCLUSIONS: Diabetes patients with severe depressive symptoms are more likely than those without depressive symptoms to report suboptimal clinician-patient communication across multiple domains of communication, especially those that involve more interactive and "patient-centered" communication. Further investigation of this relationship may uncover explanatory mechanisms and help guide interventions for improving care for both conditions. PMID- 18388841 TI - Validation and standardization of the Generalized Anxiety Disorder Screener (GAD 7) in the general population. AB - BACKGROUND: The 7-item Generalized Anxiety Disorder Scale (GAD-7) is a practical self-report anxiety questionnaire that proved valid in primary care. However, the GAD-7 was not yet validated in the general population and thus far, normative data are not available. OBJECTIVES: To investigate reliability, construct validity, and factorial validity of the GAD-7 in the general population and to generate normative data. RESEARCH DESIGN: Nationally representative face-to-face household survey conducted in Germany between May 5 and June 8, 2006. SUBJECTS: Five thousand thirty subjects (53.6% female) with a mean age (SD) of 48.4 (18.0) years. MEASURES: The survey questionnaire included the GAD-7, the 2-item depression module from the Patient Health Questionnaire (PHQ-2), the Rosenberg Self-Esteem Scale, and demographic characteristics. RESULTS: Confirmatory factor analyses substantiated the 1-dimensional structure of the GAD-7 and its factorial invariance for gender and age. Internal consistency was identical across all subgroups (alpha = 0.89). Intercorrelations with the PHQ-2 and the Rosenberg Self Esteem Scale were r = 0.64 (P < 0.001) and r = -0.43 (P < 0.001), respectively. As expected, women had significantly higher mean (SD) GAD-7 anxiety scores compared with men [3.2 (3.5) vs. 2.7 (3.2); P < 0.001]. Normative data for the GAD-7 were generated for both genders and different age levels. Approximately 5% of subjects had GAD-7 scores of 10 or greater, and 1% had GAD-7 scores of 15 or greater. CONCLUSIONS: Evidence supports reliability and validity of the GAD-7 as a measure of anxiety in the general population. The normative data provided in this study can be used to compare a subject's GAD-7 score with those determined from a general population reference group. PMID- 18388843 TI - Medical care costs one year after identification of hyperglycemia below the threshold for diabetes. AB - OBJECTIVE: To estimate the resource utilization and medical costs of patients with impaired fasting glucose (IFG), impaired glucose tolerance (IGT), or both, in a real-world clinical setting. METHODS: We used fasting and random glucose test results and a previously validated predictive equation to identify glycemic status in 26,111 nondiabetic patients, assigning them to categories of normoglycemia, isolated IFG (I-IFG), isolated IGT (I-IGT), or IFG with possible IGT (IFG/IGT). We then calculated and compared mean annual medical resource utilization and age/sex-adjusted costs over the ensuing 12-month period. RESULTS: I-IGT patients incurred significantly greater age- and sex-adjusted total costs in the observation year compared with normoglycemic and I-IFG patients (both comparisons, P < 0.001). IFG/IGT patients also had significantly greater age- and sex-adjusted total costs in the observation year compared with normoglycemic and I-IFG patients (P < 0.001, both comparisons). In both cases, the differences were driven by significantly greater inpatient costs-20.3% of patients with I-IGT and 17.1% with IFG/IGT were hospitalized during the observation year, whereas approximately 12% of normoglycemic and I-IFG patients had an admission (all comparisons, P < 0.001). CONCLUSIONS: Abnormal glucose tolerance, in particular, IGT, is associated with excess medical care costs relative to normoglycemia. Preventing progression to diabetes, when costs are known to be dramatically greater, would likely provide substantial economic benefit. More research is needed to determine the prevalence of hyperglycemia-related complications at elevated glucose levels below the diabetic threshold and the associated costs of those complications. PMID- 18388844 TI - The association between patient satisfaction and quality of life in Chinese lung and liver cancer patients. AB - BACKGROUND: Patient satisfaction has been accorded increased importance in evaluating the quality of health care. However, only a few studies have examined factors associated with patient satisfaction in cancer populations, with no data currently available in Chinese oncology populations. OBJECTIVES: We sought to examine the longitudinal course of the relationship between patient satisfaction and quality of life (QoL) in Chinese cancer patients. METHODS: A total of 253 liver cancer patients and 334 lung cancer patients were assessed during their first outpatient visit (baseline) and at 2 follow-up interviews (FU1 and FU2). QoL was measured by the Chinese version of the Functional Assessment of Cancer Therapy-General Scale [FACT-G (Ch)]. Patient satisfaction was assessed by the Nine-Item Chinese Patient Satisfaction Questionnaire (ChPSQ-9) and the cognitive subscale of the Medical Interview Satisfaction Scale (MISS-Cog). Linear mixed effects (LME) models were fitted to identify predictors of patient satisfaction and QoL. RESULTS: Results of LME analyses showed no significant between-group differences in psychosocial, satisfaction, and QoL measures over time (all P > 0.05). Age [beta = -0.001, 95% confidence interval (CI): -0.03 to 0.00, P < 0.05] and optimism (beta = 0.20, 95% CI: 0.10 to 0.30, P < 0.001) predicted MISS-Cog and ChPSQ-9, respectively. After adjusting for sociodemographic and psychosocial variables, only ChPSQ-9 (beta = 0.12, 95% CI: 0.06 to 0.18, P < 0.001) predicted QoL scores. No significant association between MISS-Cog and QoL was found. CONCLUSIONS: General emotional support from health professionals (ChPSQ-9) was a more effective predictor of QoL than was a measure of informational support (MISS Cog) among these patients. Because expectations of clinical care likely vary by culture, and nature and stage of disease, measures of patient satisfaction that address both informational and care expectations are important. PMID- 18388842 TI - Variation in treatment preferences and care goals among older patients with diabetes and their physicians. AB - BACKGROUND: Older persons with diabetes are heterogeneous with respect to life expectancy and frailty, and new guidelines recommend individualizing care. OBJECTIVES: (1) To describe variation in the preferences of older patients with diabetes regarding aggressiveness of glycemic control and avoiding diabetic complications. (2) To determine correlates of patient preferences and physician treatment goals. (3) To assess whether physicians' goals were consistent with their patients' preferences. RESEARCH DESIGN: Cross-sectional surveys and chart reviews in urban academic clinics. SUBJECTS: : Four hundred seventy-three patients with diabetes age 65 or older and 64 physicians. MEASURES: Patient preferences (utilities on a scale from 0 to 1, 0 = death, 1 = perfect health) for diabetic complications and intensity of treatment, and physician target treatment goals and ratings of aggressiveness of approach. RESULTS: Eighty percent of the patients were African American, 63% were women, average age was 73.7 +/- 5.9 years and 26% expected to live 5 years or less. Patient preferences/utilities showed significant variation: blindness 0.39 (SD, 0.32), lower leg amputation 0.45 (0.34), conventional treatment 0.76 (0.27), and intensive insulin treatment 0.64 (0.32). Physicians' hemoglobin A1c goal was < or =7% in 69% of patients. Greater estimated patient life expectancy was consistently associated with higher patient utilities and was associated with physicians' willingness to use aggressive treatments. Physicians' treatment goals and approaches were associated with patients' utilities for treatment. CONCLUSIONS: Older patients vary greatly in their preferences regarding diabetic complications and treatments. Acknowledging patient preferences, along with life goals and prognostic data, may improve quality of treatment decisions. PMID- 18388845 TI - Measuring clinically significant chemotherapy-related toxicities using Medicare claims from Cancer and Leukemia Group B (CALGB) trial participants. AB - BACKGROUND: Because the elderly are underrepresented on clinical trials, physicians have few sources of information to estimate the risks (ie, toxicities) and benefits of chemotherapy administration to the elderly. OBJECTIVE: Our goal was to determine whether the standard measures of toxicity used in clinical trials could be captured from observational Medicare claims data. METHODS: We identified 175 elderly clinical trial patients treated on 2 Cancer and Leukemia Group B (CALGB) trials (9344, adjuvant breast study and 9730, advanced lung cancer study) and merged participants' CALGB data with their Medicare data. From CALGB data, we identified the most frequent Extended Clinical Toxicity Critieria grade III/IV toxicities. We reviewed diagnostic and procedure codes from Medicare manuals, developed algorithms to measure the toxicities, and then finalized the algorithms after empiric review of patients' codes. We compared results of Medicare algorithms to gold standard CALGB toxicity information to calculate test characteristics. RESULTS: CALGB data documented that 15 grade III/IV chemotherapy related toxicities occurred in > or =3% of the 175 patients: white blood cell, hemoglobin, platelets, anorexia, nausea, vomiting, diarrhea, stomatitis, sensory neuropathy, motor neuropathy, motor or sensory neuropathy, dyspnea, hyperglycemia, infection, and malaise. Vomiting was the only toxicity identified by the Medicare-based algorithm with a sensitivity, specificity, and area under the receiver operator curve of > or =80%. CONCLUSIONS: The results of this preliminary study suggest that Medicare diagnostic and procedure codes may be of only limited value in measuring clinically significant chemotherapy-related toxicities in elderly Medicare beneficiaries. Future research includes confirming these findings in a larger and more diverse sample. PMID- 18388846 TI - Dual use of Veterans Affairs services and use of recommended ambulatory care. AB - BACKGROUND: Use of more than one health care system to obtain care is common among adults receiving care within the Veterans Affairs (VA) medical system. It is not known what effect using care from multiple sources has on the quality of care patients receive. OBJECTIVES: To examine whether use of recommended ambulatory care services differs between exclusive and dual VA users. METHODS: Cross-sectional analysis of the 2004 Behavior Risk Factor Surveillance System, a nationally-representative survey of community-dwelling adults aged 18 years or older. Our outcome measures were self-reported use of 18 recommended services for cancer prevention, cardiovascular risk reduction, diabetes management, and infectious disease prevention. We used multivariable logistic regression to examine the association between exclusive and dual VA use and use of recommended ambulatory services. RESULTS: There were 3470 exclusive VA users and 4523 dual VA users. Dual users were significantly more likely to be older and white, have higher incomes, have graduated from college, and be insured when compared with exclusive VA users. In unadjusted analyses, dual users received higher rates of recommended services. After adjustment for patient characteristics, use of recommended services was largely similar among exclusive and dual VA users. Exclusive VA users reported 14% greater use of breast cancer screening and 10% greater use of cholesterol monitoring among patients with hypercholesterolemia, and 6% lower use of prostate cancer screening and 7% lower use of influenza vaccination. CONCLUSIONS: After adjustment for patient characteristics, exclusive and dual VA users reported similar rates of recommended ambulatory service use. PMID- 18388847 TI - Assessing the quality of transitional care: further applications of the care transitions measure. AB - BACKGROUND: The Institute of Medicine has identified care transitions as a priority area for performance measurement. OBJECTIVES: To examine the performance of the Care Transitions Measure (CTM) in more diverse populations and to introduce a 3-item CTM. RESEARCH DESIGN: Cross-sectional study with purposive sampling of traditionally underserved populations. Confirmatory factor analyses, internal consistency reliability analyses, and differential item function tests were performed to explore the stability and performance of the 15-item CTM. Regression assessed the ability of the 3-item CTM to predict the 15-item CTM total score. Analysis of variance tests were conducted to explore CTM performance in different populations with respect to health and demographics. SUBJECTS: A total of 225 patients age 18-90 who were hospitalized in the past 12 months and were African American, Hispanic American, or rural-dwelling. MEASURES: CTM-15, CTM-3, age, gender, education, and health status. RESULTS: Mplus confirmatory factor analysis supported the CTM-15 factor structure in more diverse population (Comparative Fit Index [CFI] = 0.954). The 3-item CTM explained 88% of the variance in the 15-item CTM score. Differential item function analysis did not reveal any differential item difficulty by age, gender, education, self-rated health, or group (African American, Hispanic American, and rural-dwelling). CONCLUSIONS: Following endorsement by National Quality Forum, findings support use of the CTM in national public reporting efforts. The 3-item CTM closely approximates the 15-item instrument and may be attractive to purchasers and health care organizations that want to assess quality in this area while minimizing cost and response burden. PMID- 18388848 TI - Does having a regular primary care clinician improve quality of preventive care for young children? AB - OBJECTIVE: This study examines whether having a regular clinician for preventive care is associated with quality of care for young children, as measured by interpersonal quality ratings and content of anticipatory guidance. DATA SOURCE: The National Survey of Early Childhood Health (NSECH), a nationally representative parent survey of health care quality for 2068 young US children fielded by the National Center for Health Statistics (NCHS). STUDY DESIGN: Bivariate and multivariate analyses evaluate associations between having a regular clinician for well child care and interpersonal quality, the content of anticipatory guidance, and timely access to care. PRINCIPAL FINDINGS: In bivariate analysis, parents of children with a regular clinician for preventive care reported slightly higher interpersonal quality (69 vs. 65 on a 0-100 scale, P = 0.01). Content of anticipatory guidance received was slightly greater for children with a regular clinician (82 vs. 80 on a 0-100 scale, P = 0.03). In bivariate analysis, a regular clinician was associated with interpersonal quality only among African American and Hispanic children. In multivariate analyses, controlling for factors that could independently influence self-reports of experiences with care, interpersonal quality but not anticipatory guidance content was higher for children with a regular clinician. CONCLUSIONS: Having a regular primary care clinician is embraced in pediatrics, although team care among physicians is also widely practiced. For young children, having a regular clinician is associated with modest gains in interpersonal quality and no differences in content of anticipatory guidance. The benefit of having a regular clinician may primarily occur in interpersonal quality for subgroups of young children. PMID- 18388849 TI - The role of hospital profit status in pediatric spleen injury management. AB - BACKGROUND: Evidence suggests that over 90% of pediatric splenic injuries can be successfully managed nonoperatively. Although pediatric hospitals have rapidly adopted nonoperative management, variability exists among other types of hospitals. OBJECTIVES: We tested 2 a priori hypotheses: (1) spleen-injured children are more likely to receive splenectomy in for-profit hospitals than in not-for-profit hospitals; and (2) hospital charges for spleen cases are greater in for-profit hospital than in not-for-profit hospitals. RESEARCH DESIGN: Multivariable regression was performed with data from the Kid's Inpatient Database (KID) for years 2000 and 2003, controlling for patient and hospital characteristics. Children (0-18 years) hospitalized with a blunt traumatic (noniatrogenic) spleen injury in any of the states participating in KID (N = 5061), including adult and pediatric hospitals. Main outcome measures were splenectomy and hospital charges. RESULTS: A total of 756 children (14.9%) received splenectomies within 1 day of arrival. Splenectomy was found to be more likely among children treated at for-profit hospitals [odds ratio (OR), 1.75; 95% confidence interval (CI), 1.14-2.67] than among children treated in not-for profit general hospitals. Splenectomies were much less common in children's hospitals (OR, 0.14; 95% CI, 0.05-0.41) than in not-for-profit general hospitals. Hospital charges for all spleen-injured children (regardless of treatment) were significantly greater in for-profit hospitals than in not-for-profit hospitals. CONCLUSIONS: For-profit hospitals seem to be trailing not-for-profit hospitals in the adoption of spleen-conserving management practices. The cost of caring for a child with a splenic injury also seems greater at for-profit hospitals, regardless of management path (ie, splenectomy vs. nonoperative management). PMID- 18388850 TI - Have HMOs broadened their hospital networks?: Changes in HMO hospital networks in California, 1999-2003. AB - OBJECTIVE: This study uses empirical data to study changes in the structure and use of HMO hospital provider networks in California. STUDY DESIGN: Data from California HMOs are used to test whether there have been structural changes in HMO size, geographic coverage, hospital network design, and patient channeling for inpatient care over the period 1999-2003. METHODS: Three different measures of HMO network breadth, access, and channeling were analyzed between 1999 and 2003. Actual patient admission data linked to health plan code variables are used to identify inpatient hospital discharges covered by each HMO in California and to which hospital each HMO sends its patients in each year between 1999 and 2003. RESULTS: Despite consolidation in the total number of HMOs, the share of all hospital admissions accounted for by HMOs remained substantial. In terms of network breadth, there were minimal changes over time in the percent of available hospitals included in HMO networks. There was a slight increase in distance traveled for HMO' patient who were admitted, the opposite of what would be expected if networks were being broadened. Finally, channeling, as measured by the concentration of a payers' patients within its network hospitals did not change significantly. CONCLUSIONS: We found little evidence that there have been systematic changes in either the structure or use of HMO hospital networks in California between 1999 and 2003, suggesting that these factors played a limited role in explaining the return of growth in health care costs. PMID- 18388851 TI - [The two faces of reactive oxygen species]. AB - Oxidative stress has been implicated in playing a crucial role in aging and in the pathogeneses of a number of diseases, including neurodegenerative disorders such as Alzheimer's disease. Oxidative stress occurs due to an imbalance in prooxidant and antioxidant levels. Reactive oxygen species (ROS) are highly reactive and may modify and inactivate proteins, lipids, DNA, and RNA and induce cellular dysfunctions. To prevent free radical-induced cellular damage, the organism has developed a defense mechanism, the antioxidative system. This system includes antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSHPx), and glutathione reductase (GSSGR) and low molecular antioxidants such as glutathion and plasma proteins. Glutathion plays a key role in maintaining the physiological balance between prooxidants and antioxidants. Plasma proteins can inhibit ROS generation and lipid peroxidation by chelating free transition metals. The major exogenous antioxidants are vitamins E, C, and A. PMID- 18388852 TI - [Cancer chemopreventive agents: glucosinolates and their decomposition products in white cabbage (Brassica oleracea var. capitata)]. AB - A number of recent epidemiological studies have indicated that high intake of white cabbage may be associated with a lower risk of neoplastic diseases such as cancer of the pancreas, breast, prostate, stomach, and lungs. The anticarcinogenic activity is related to the presence of biologically active components in this vegetable. The chemopreventive effects of cabbage may be connected with modulation of the activity of phase I and II detoxification enzymes and other mechanisms triggered by glucosinolates and products of their decomposition, which are formed as a result of hydrolysis catalyzed by the enzyme myrosinase. The products of glucosinolate decomposition influence a number of cellular processes through the regulation of transcription factor levels, signaling pathways, the cell cycle, and apoptosis. The beneficial activities and especially the chemopreventive effects of the compounds present in cabbage point to the necessity of formulating scientifically based dietary recommendations enabling the optimal exploitation of this vegetable in health protection. PMID- 18388853 TI - Neurological and psychiatric disorders in thyroid dysfunctions. The role of nuclear medicine: SPECT and PET imaging. AB - Thyroid dysfunctions may be accompanied by numerous neurological and psychiatric disorders. The most known is cognitive impairment and depression in hypothyroid patients, as well as an increased risk of cerebrovascular accidents. A separate, although a rare entity, is Hashimoto's encephalopathy. In hyperthyroidism there is an increased incidence of psychiatric disorders, including apathetic hyperthyroidism and hyperthyroid dementia. Functional imaging of cerebral blood flow and metabolism helped establish both global and/or regional decrease of both cerebral blood flow and metabolism in hypothyroidism, particularly in regions mediating attention, motor speed and visuospatial processing. Hypothyroid dementia may be mediated by neurocircuitry different from that in major depression. Less is known on flow/metabolism changes in hyperthyroidism. Global blood flow may be slightly increased, with regional deficits of blood flow, particular in hyperthyroid dementia. As presented above radionuclide functional imaging showed some metabolic patterns in thyroid dysfunctions, but still many issues remain unresolved. In particular little is known about the underlying pathology of cognitive impairment and depression in hypothyroidism, which may differ from ones in euthyroid patients. Also little is known about the reversibility of changes in cerebral blood flow following thyroid replacement therapy. In hyperthyroid patients functional imaging might contribute to elucidate the background of apathetic hyperthyroidism and potential different background of psychiatric complications. PMID- 18388854 TI - Role of PET in medullary thyroid carcinoma. AB - In the diagnostic assessment of medullary thyroid carcinoma (MTC), nuclear medicine imaging provides its contribution mainly in the postoperative work-up to detect residual or recurrent tumor. With respect to scintigraphy with gamma emitter radiopharmaceuticals, positron emission tomography (PET) offers interesting perspectives owing to its higher image quality, spatial resolution and speed. Moreover, the recent developments of hybrid machines allow to obtain images that simultaneously hold both anatomic (computed tomography) and functional (PET) information with great impact on diagnostic efficacy. (18)F fluoro-deoxyglucose ((18)F-FDG) is the most frequently used PET tracer in oncology. Preliminary reports of FDG-PET in MTC patients show encouraging results with a higher sensitivity in detecting local recurrent and metastatic disease when compared with single photon emission tracers. However, (18)F-FDG uptake depends on lesion size and to some extent on the grade of differentiation and biologic aggressiveness of the tumor; so FDG-PET seems useful mainly in patients with very high calcitonin levels and high progression rate. Like other neuroendocrine tumors, MTC is characterized by the presence of amine uptake mechanism and/or peptide receptors at the cell membrane allowing the clinical use of specific radiopharmaceuticals that reflect the different metabolic pathways of MTC, and in particular the synthesis, storage and release of hormones ((18)F dihydroxyphenilalanine, (18)F-DOPA and (18)F-fluorodopamine, (18)F-FDA) and the expression of receptors ((68)Ga-labeled somatostatin analogs). These tracers are currently under investigation and will further improve the diagnostic approach of MTC. PMID- 18388855 TI - Nuclear medicine in endocrinology. PMID- 18388856 TI - Activation of TRPP2 through mDia1-dependent voltage gating. AB - The TRPP2 cation channel is directly responsible for approximately 15% of all cases of autosomal dominant polycystic kidney disease. However, the mechanisms underlying fundamental properties of TRPP2 regulation, such as channel gating and activation, are unknown. We have shown that TRPP2 was activated by EGF and physically interacted with the mammalian diaphanous-related formin 1 (mDia1), a downstream effector of RhoA. Now, we show that mDia1 regulates TRPP2 by specifically blocking its activity at negative but not positive potentials. The voltage-dependent unblock of TRPP2 by mDia1 at positive potentials is mediated through RhoA-induced molecular switching of mDia1 from its autoinhibited state at negative potentials to its activated state at positive potentials. Under physiological resting potentials, EGF activates TRPP2 by releasing the mDia1 dependent block through the activation of RhoA. Our data reveal a new role of mDia1 in the regulation of ion channels and suggest a molecular basis for the voltage-dependent gating of TRP channels. PMID- 18388857 TI - Motor step size and ATP coupling efficiency of the dsDNA translocase EcoR124I. AB - The Type I restriction-modification enzyme EcoR124I is an archetypical helicase based dsDNA translocase that moves unidirectionally along the 3'-5' strand of intact duplex DNA. Using a combination of ensemble and single-molecule measurements, we provide estimates of two physicochemical constants that are fundamental to a full description of motor protein activity-the ATP coupling efficiency (the number of ATP consumed per base pair) and the step size (the number of base pairs transported per motor step). Our data indicate that EcoR124I makes small steps along the DNA of 1 bp in length with 1 ATP consumed per step, but with some uncoupling of the ATPase and translocase cycles occurring so that the average number of ATP consumed per base pair slightly exceeds unity. Our observations form a framework for understanding energy coupling in a great many other motors that translocate along dsDNA rather than ssDNA. PMID- 18388858 TI - Arabidopsis COP1 shapes the temporal pattern of CO accumulation conferring a photoperiodic flowering response. AB - The transcriptional regulator CONSTANS (CO) promotes flowering of Arabidopsis under long summer days (LDs) but not under short winter days (SDs). Post translational regulation of CO is crucial for this response by stabilizing the protein at the end of a LD, whereas promoting its degradation throughout the night under LD and SD. We show that mutations in CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1), a component of a ubiquitin ligase, cause extreme early flowering under SDs, and that this is largely dependent on CO activity. Furthermore, transcription of the CO target gene FT is increased in cop1 mutants and decreased in plants overexpressing COP1 in phloem companion cells. COP1 and CO interact in vivo and in vitro through the C-terminal region of CO. COP1 promotes CO degradation mainly in the dark, so that in cop1 mutants CO protein but not CO mRNA abundance is dramatically increased during the night. However, in the morning CO degradation occurs independently of COP1 by a phytochrome B-dependent mechanism. Thus, COP1 contributes to day length perception by reducing the abundance of CO during the night and thereby delaying flowering under SDs. PMID- 18388859 TI - Akt- and Foxo1-interacting WD-repeat-FYVE protein promotes adipogenesis. AB - We have previously identified a protein, consisting of seven WD-repeats, forming a putative beta-propeller, and an FYVE domain, ProF, which is highly expressed in 3T3-L1 cells, a cell line that can be differentiated into adipocytes. We recently found ProF to interact with the kinases Akt and protein kinase Czeta. Here we demonstrate that ProF is a positive regulator of adipogenesis. Knockdown of ProF by RNA interference leads to decreased adipocyte differentiation. This is shown by reduced lipid accumulation, decreased expression of the differentiation markers PPARgamma and C/EBPalpha, and reduced glucose uptake in differentiated cells. Furthermore, ProF overexpression leads to increased adipogenesis. ProF binds to the transcription factor Foxo1 (Forkhead box O1), a negative regulator of insulin action and adipogenesis, and facilitates the phosphorylation and thus inactivation of Foxo1 by Akt. Additionally, dominant-negative Foxo1 restores adipogenesis in ProF knockdown cells. Thus, ProF modulates Foxo1 phosphorylation by Akt, promoting adipocyte differentiation. Furthermore, ProF might be involved in metabolic disorders such as diabetes. PMID- 18388860 TI - Mechanism of intercellular molecular exchange in heterocyst-forming cyanobacteria. AB - Heterocyst-forming filamentous cyanobacteria are true multicellular prokaryotes, in which heterocysts and vegetative cells have complementary metabolism and are mutually dependent. The mechanism for metabolite exchange between cells has remained unclear. To gain insight into the mechanism and kinetics of metabolite exchange, we introduced calcein, a 623-Da fluorophore, into the Anabaena cytoplasm. We used fluorescence recovery after photobleaching to quantify rapid diffusion of this molecule between the cytoplasms of all the cells in the filament. This indicates nonspecific intercellular channels allowing the movement of molecules from cytoplasm to cytoplasm. We quantify rates of molecular exchange as filaments adapt to diazotrophic growth. Exchange among vegetative cells becomes faster as filaments differentiate, becoming considerably faster than exchange with heterocysts. Slower exchange is probably a price paid to maintain a microaerobic environment in the heterocyst. We show that the slower exchange is partly due to the presence of cyanophycin polar nodules in heterocysts. The phenotype of a null mutant identifies FraG (SepJ), a membrane protein localised at the cell-cell interface, as a strong candidate for the channel-forming protein. PMID- 18388861 TI - Mus81 is essential for sister chromatid recombination at broken replication forks. AB - Recombination is essential for the recovery of stalled/collapsed replication forks and therefore for the maintenance of genomic stability. The situation becomes critical when the replication fork collides with an unrepaired single strand break and converts it into a one-ended double-strand break. We show in fission yeast that a unique broken replication fork requires the homologous recombination (HR) enzymes for cell viability. Two structure-specific heterodimeric endonucleases participate in two different resolution pathways. Mus81/Eme1 is essential when the sister chromatid is used for repair; conversely, Swi9/Swi10 is essential when an ectopic sequence is used for repair. Consequently, the utilization of these two HR modes of resolution mainly relies on the ratio of unique and repeated sequences present in various eukaryotic genomes. We also provide molecular evidence for sister recombination intermediates. These findings demonstrate that Mus81/Eme1 is the dedicated endonuclease that resolves sister chromatid recombination intermediates during the repair of broken replication forks. PMID- 18388862 TI - Functioning of the dimeric GABA(B) receptor extracellular domain revealed by glycan wedge scanning. AB - The G-protein-coupled receptor (GPCR) activated by the neurotransmitter GABA is made up of two subunits, GABA(B1) and GABA(B2). GABA(B1) binds agonists, whereas GABA(B2) is required for trafficking GABA(B1) to the cell surface, increasing agonist affinity to GABA(B1), and activating associated G proteins. These subunits each comprise two domains, a Venus flytrap domain (VFT) and a heptahelical transmembrane domain (7TM). How agonist binding to the GABA(B1) VFT leads to GABA(B2) 7TM activation remains unknown. Here, we used a glycan wedge scanning approach to investigate how the GABA(B) VFT dimer controls receptor activity. We first identified the dimerization interface using a bioinformatics approach and then showed that introducing an N-glycan at this interface prevents the association of the two subunits and abolishes all activities of GABA(B2), including agonist activation of the G protein. We also identified a second region in the VFT where insertion of an N-glycan does not prevent dimerization, but blocks agonist activation of the receptor. These data provide new insight into the function of this prototypical GPCR and demonstrate that a change in the dimerization interface is required for receptor activation. PMID- 18388863 TI - A synthetic lethal siRNA screen identifying genes mediating sensitivity to a PARP inhibitor. AB - Inhibitors of poly (ADP-ribose)-polymerase-1 (PARP) are highly lethal to cells with deficiencies in BRCA1, BRCA2 or other components of the homologous recombination pathway. This has led to PARP inhibitors entering clinical trials as a potential therapy for cancer in carriers of BRCA1 and BRCA2 mutations. To discover new determinants of sensitivity to these drugs, we performed a PARP inhibitor synthetic lethal short interfering RNA (siRNA) screen. We identified a number of kinases whose silencing strongly sensitised to PARP inhibitor, including cyclin-dependent kinase 5 (CDK5), MAPK12, PLK3, PNKP, STK22c and STK36. How CDK5 silencing mediates sensitivity was investigated. Previously, CDK5 has been suggested to be active only in a neuronal context, but here we show that CDK5 is required in non-neuronal cells for the DNA-damage response and, in particular, intra-S and G(2)/M cell-cycle checkpoints. These results highlight the potential of synthetic lethal siRNA screens with chemical inhibitors to define new determinants of sensitivity and potential therapeutic targets. PMID- 18388865 TI - The unanswered question: what is the determinant of S-1 pharmacokinetics? PMID- 18388864 TI - The FoxO3a gene is a key negative target of canonical Notch signalling in the keratinocyte UVB response. AB - Notch signalling has an important role in skin homeostasis, promoting keratinocyte differentiation and suppressing tumorigenesis. Here we show that this pathway also has an essential anti-apoptotic function in the keratinocyte UVB response. Notch1 expression and activity are significantly induced, in a p53 dependent manner, by UVB exposure of primary keratinocytes as well as intact epidermis of both mouse and human origin. The apoptotic response to UVB is increased by deletion of the Notch1 gene or down-modulation of Notch signalling by pharmacological inhibition or genetic suppression of 'canonical' Notch/CSL/MAML1-dependent transcription. Conversely, Notch activation protects keratinocytes against apoptosis through a mechanism that is not linked to Notch induced cell cycle withdrawal or NF-kappaB activation. Rather, transcription of FoxO3a, a key pro-apoptotic gene, is under direct negative control of Notch/HERP transcription in keratinocytes, and upregulation of this gene accounts for the increased susceptibility to UVB of cells with suppressed Notch signalling. Thus, the canonical Notch/HERP pathway functions as a protective anti-apoptotic mechanism in keratinocytes through negative control of FoxO3a expression. PMID- 18388866 TI - Emerging evidence of the impact of kidney disease on drug metabolism and transport. AB - Several lines of emerging evidence indicate that kidney disease differentially affects uptake and efflux transporters and metabolic enzymes in the liver and gastrointestinal (GI) tract, and uremic toxins have been implicated as the cause. In patients with kidney disease, even drugs that are eliminated by nonrenal transport and metabolism could lead to important unintended consequences if they are administered without dose adjustment for reduced renal function. This is particularly so in the case of drugs with narrow therapeutic windows and may translate into clinically significant variations in exposure and response. PMID- 18388867 TI - Optical molecular imaging: from single cell to patient. AB - Optical imaging is an emerging field with a wide range of biomedical research and clinical applications, both current and future. It comprises several classes of techniques that are capable of providing information at the molecular, cellular, tissue, and whole-animal levels. These techniques match well with emerging genomic and proteomic technologies that enable development of optical "probes," as well as with nanotechnologies for multifunctional imaging and drug delivery. These advances have enormous potential to accelerate drug discovery/development by providing predictive information on mechanisms of action and biological responses. PMID- 18388868 TI - Toward personalized therapy for smoking cessation: a randomized placebo controlled trial of bupropion. PMID- 18388870 TI - Inflammation and altered drug clearance in cancer: transcriptional repression of a human CYP3A4 transgene in tumor-bearing mice. AB - A tumor-associated inflammatory response has recently been found to contribute to the considerable interindividual variability in cytotoxic drug clearance seen in cancer patients. Circulating inflammatory markers, such as C-reactive protein (CRP) and interleukin-6 (IL-6), correlate with excessive drug toxicity caused by reduced CYP3A4-mediated metabolism. This article outlines the use of a transgenic mouse model of human CYP3A4 regulation to demonstrate that extrahepatic tumors elicit an inflammatory response, leading to transcriptional repression of the CYP3A4 gene as well as of other drug clearance pathways. PMID- 18388871 TI - Genetic influences in the variation in renal clearance of nicotine and cotinine. AB - Nicotine and its proximate metabolite cotinine are eliminated in part by renal clearance. These compounds are filtered, secreted, and reabsorbed, and the resultant renal clearances are quite variable among individuals and are highly influenced by urine pH. In this study of 139 pairs of twins, we have estimated the genetic and environmental contributions to total renal clearance and net secretory/reabsorptive clearance of nicotine and cotinine. At uncontrolled urine pH both nicotine and cotinine undergo net reabsorption. Additive genetic factors were not important contributors to the variation in total renal clearance of nicotine but played a relatively more substantial role in accounting for the variation in total renal clearance of cotinine (43% of variance). Variations in glomerular filtration rate and the net secretory/reabsorptive clearance of nicotine and cotinine were largely influenced by nonadditive genetic influences (41.5-61% of variance). Earlier research has shown that renal secretory clearance of drugs can be highly heritable, presumably related to genetic variation in transporters. Our study suggests that the renal clearance of drugs that undergo extensive renal reabsorption can be substantially influenced by nonadditive genetic and/or shared environmental factors. PMID- 18388872 TI - Mathematics for understanding disease. AB - The application of mathematical models to reflect the organization and activity of biological systems can be viewed as a continuum of purpose. The far left of the continuum is solely the prediction of biological parameter values, wherein an understanding of the underlying biological processes is irrelevant to the purpose. At the far right of the continuum are mathematical models, the purposes of which are a precise understanding of those biological processes. No models in present use fall at either end of the continuum. Without question, however, the emphasis in regards to purpose has been on prediction, e.g., clinical trial simulation and empirical disease progression modeling. Clearly the model that ultimately incorporates a universal understanding of biological organization will also precisely predict biological events, giving the continuum the logical form of a tautology. Currently that goal lies at an immeasurable distance. Nonetheless, the motive here is to urge movement in the direction of that goal. The distance traveled toward understanding naturally depends upon the nature of the scientific question posed with respect to comprehending and/or predicting a particular disease process. A move toward mathematical models implies a move away from static empirical modeling and toward models that focus on systems biology, wherein modeling entails the systematic study of the complex pattern of organization inherent in biological systems. PMID- 18388873 TI - Development of translational pharmacokinetic-pharmacodynamic models. AB - Contemporary models in the field of pharmacokinetic-pharmacodynamic (PK-PD) modeling often incorporate the fundamental principles of capacity limitation and operation of turnover processes to describe the time course of pharmacological effects in mechanistic terms. This permits the identification of drug- and system specific factors that govern drug responses. There is considerable interest in utilizing mechanism-based PK-PD models in translational pharmacology, whereby in silico, in vitro, and preclinical data may be effectively coupled with relevant models to streamline the discovery and development of new therapeutic agents. These translational PK-PD models form the subject of this review. PMID- 18388874 TI - Lazarus's gate: challenges and potential of epigenetic reprogramming of somatic cells. AB - The past year has seen tremendous advances in epigenetic reprogramming of somatic cells. Direct genesis of pluripotent stem cells, in contrast to earlier somatic cell nuclear transfer (SCNT) techniques, removes significant ethical and regulatory concerns regarding the utilization of human oocytes and zygotes, and represents a significant step toward the development of nonxenogeneic production methods. While significant technical hurdles remain, this and related technologies are enabling new approaches toward clinical treatments, basic research and diagnostics, and drug evaluation. PMID- 18388875 TI - Mice as clinically relevant models for the study of cytochrome P450-dependent metabolism. AB - The cytochrome P450 (CYP) gene superfamily comprises a large group of hemoproteins with diverse functions in steroid, lipid, and xenobiotic metabolism. The human genome is estimated to contain 57 genes that encode functional CYP proteins, a number of which are important for the metabolism of foreign chemicals, including carcinogens and most therapeutic drugs. Given that metabolic interactions are a major source of adverse drug interactions, a comprehensive understanding of CYP function is critically important for the development and safe clinical application of drugs. While some cross-species genetic conservation of CYPs exists, drug metabolism can differ between humans and other mammalian species. The development of humanized mice that replicate many aspects of human drug metabolism has provided invaluable experimental models that circumvent this limitation to a considerable degree. This brief review focuses on the value and limitations of mouse models for the study of drug metabolism in humans. PMID- 18388877 TI - The effect of gemfibrozil on repaglinide pharmacokinetics persists for at least 12 h after the dose: evidence for mechanism-based inhibition of CYP2C8 in vivo. AB - Repaglinide is metabolized by cytochrome P450 (CYP) 2C8 and 3A4. Gemfibrozil has the effect of increasing the area under the concentration-time curve (AUC) of repaglinide eightfold. We studied the effect of dosing interval on the extent of the gemfibrozil-repaglinide interaction. In a randomized five-phase crossover study, 10 healthy volunteers ingested 0.25 mg repaglinide, with or without gemfibrozil pretreatment. Plasma repaglinide, gemfibrozil, their metabolites, and blood glucose were measured. When the last dose of 600 mg gemfibrozil was ingested simultaneously with repaglinide, or 3, 6, or 12 h before, it increased the AUC(0-infinity) of repaglinide 7.0-, 6.5-, 6.2- and 5.0-fold, respectively (P < 0.001). The peak repaglinide concentration increased approximately twofold (P < 0.001), and the half-life was prolonged from 1.2 h to 2-3 h (P < 0.001) during all the gemfibrozil phases. The drug interaction effects persisted at least 12 h after gemfibrozil was administered, although plasma gemfibrozil and gemfibrozil 1 O-beta-glucuronide concentrations were only 5 and 10% of their peak values, respectively. The long-lasting interaction is likely caused by mechanism-based inhibition of CYP2C8 by gemfibrozil glucuronide. PMID- 18388880 TI - Development of procedures for early screening of smoking cessation medications in humans. AB - Candidate medications for smoking cessation may be screened more efficiently if initial evaluations in humans combine the practical advantages of laboratory studies with the clinical validity of clinical trials, such as by increasing participants' "quit motivation" during brief testing. We manipulated "intrinsic" quit motivation by recruiting smokers who either did intend to quit soon ("treatment seekers," N = 47) or did not ("nonseekers," N = 93), and "extrinsic" quit motivation by providing or not providing reinforcement for abstinence ($12/day). All the subjects smoked as they would usually do during weeks 1 and 3, and tried to quit during weeks 2 and 4 using either a nicotine patch (21 mg) or a placebo patch, in accordance with the crossover design of the study. The nicotine patch increased abstinence in treatment seekers but not in nonseekers. Reinforcement had a main effect on abstinence but did not moderate the effects of the nicotine patch or treatment-seeking status. Intrinsic, but not extrinsic, quit motivation of participants may enhance the validity of brief tests of medication efficacy for smoking cessation. PMID- 18388879 TI - Effect of APOE genotype on lipid levels in patients with coronary heart disease during a 3-week inpatient rehabilitation program. AB - It has been suggested that the apolipoprotein E (APOE) genotype modifies the effect of dietary and pharmacological interventions for lowering lipid levels. We wanted to determine whether APOE genotyping information would be useful in making lipid-lowering treatment decisions in clinical practice. We included 981 patients with coronary heart disease (CHD) enrolled in an inpatient 3-week standardized rehabilitation program. Of these, 555 (57%) patients received continued statin therapy and 232 (24%) patients received newly initiated statin therapy. Dietary intervention was part of the program only for 194 (20%) patients. Total cholesterol (TC) and low-density lipoprotein cholesterol (LDLC) levels decreased in all the groups of patients during rehabilitation. The decreases were less pronounced among the APOE E2 carriers. However, the observed variation among the groups with respect to reduction of lipid levels was accounted for mainly by the initial lipid levels (30-47%) and only marginally on the APOE genotype (1%) . We therefore found no evidence that APOE genotyping will be useful in guiding dietary or pharmacological lipid-lowering treatment decisions. PMID- 18388881 TI - Models for plasma glucose, HbA1c, and hemoglobin interrelationships in patients with type 2 diabetes following tesaglitazar treatment. AB - Pharmacokinetic (PK) pharmacodynamic (PD) modeling was applied to understand and quantitate the interplay between tesaglitazar (a peroxisome proliferator activated receptor alpha/gamma agonist) exposure, fasting plasma glucose (FPG), hemoglobin (Hb), and glycosylated hemoglobin (HbA1c) in type 2 diabetic patients. Data originated from a 12-week dose-ranging study with tesaglitazar. The primary objective was to develop a mechanism-based PD model for the FPG-HbA1c relationship. The secondary objective was to investigate possible mechanisms for the tesaglitazar effect on Hb. Following initiation of tesaglitazar therapy, time to new FPG steady state was approximately 9 weeks, and tesaglitazar potency in females was twice that in males. The model included aging of red blood cells (RBCs) using a transit compartment approach. The RBC life span was estimated to 135 days. The transformation from RBC to HbA1c was modeled as an FPG-dependent process. The model indicated that the tesaglitazar effect on Hb was caused by hemodilution of RBCs. PMID- 18388882 TI - Integrating family medicine and pharmacy to advance primary care therapeutics. AB - The prevalence of suboptimal prescribing of medications is well documented. Patients are often undertreated or not offered therapeutic treatments that are likely to confer benefit. As a result, drug-related hospital admissions are common and often preventable. Improvements to the health-care system are clearly needed in order to maximize the benefits that can be derived from medications. Many countries are changing their primary health-care systems to improve the quality of health-care delivery. One main transformation is the use of multidisciplinary care teams to provide care in a coordinated manner often from the same location or by using the common medical record of the patients. It has been demonstrated that pharmacists can improve prescribing, reduce health-care utilization and medication costs, and contribute to clinical improvements in many chronic medical conditions, such as cardiovascular disease, diabetes, and psychiatric illness. However, the effect of integrating a pharmacist providing general services into a primary care group has not been extensively studied. The Integrating Family Medicine and Pharmacy to Advance Primary Care Therapeutics (IMPACT) project was designed to provide a real-world demonstration of the feasibility of integrating the pharmacist into primary care office practice. This article provides a description of the IMPACT project participants; the IMPACT practice model and the concepts incorporated in its development; some initial results from the program evaluation; sustainability of the model; and some reflections on the implementation of the practice model. PMID- 18388884 TI - Application of software design principles and debugging methods to an analgesia prescription reduces risk of severe injury from medical use of opioids. AB - A prescription is a health-care program implemented by a physician or other qualified practitioner in the form of instructions that govern the plan of care for an individual patient. Although the algorithmic nature of prescriptions is axiomatic, this insight has not been applied systematically to medication safety. We used software design principles and debugging methods to create a "Patient oriented Prescription for Analgesia" (POPA), assessed the rate and extent of adoption of POPA by physicians, and conducted a statistical process control clinical trial and a subsidiary cohort analysis to evaluate whether POPA would reduce the rate of severe and fatal opioid-associated adverse drug events (ADEs). We conducted the study in a population of 153,260 hospitalized adults, 50,576 (33%) of whom received parenteral opioids. Hospitalwide, the use of POPA increased to 62% of opioid prescriptions (diffusion half-life = 98 days), while opioid-associated severe/fatal ADEs fell from an initial peak of seven per month to zero per month during the final 6 months (P < 0.0016) of the study. In the nested orthopedics subcohort, the use of POPA increased the practice of recording pain scores (94% vs. 72%, P < 0.00001) and the use of adjuvant analgesics (95% vs. 40%, P < 0.00001) and resulted in fewer opioid-associated severe ADEs than routine patient-controlled analgesia (PCA) (0% vs. 2.7%, number needed to treat (NNT) = 35, P < 0.015). The widespread diffusion of POPA was associated with a substantial hospitalwide decline in opioid-associated severe/fatal ADEs. PMID- 18388885 TI - Early and accurate prediction of Peg-IFNs/ribavirin therapy outcome in the individual patient with chronic hepatitis C by modeling the dynamics of the infected cells. AB - A novel biomathematical model that analyzes the combined alanine transaminase (ALT) and viral-load kinetics during the first month of pegylated interferon (Peg IFN) plus ribavirin (RBV) therapy was successfully applied in 90 of 97 (93%) chronic hepatitis C patients in order to compute the number of infected cells at the end of therapy (I(eot)). The I(eot) indices were lower in sustained virological responders than in relapsers (RELs) and nonresponders (NRs) (median values: 31 vs. 2,190 vs. 1,090,000; P < 0.001), and were independently associated with treatment outcomes (P = 0.003). A threshold of 250 I(eot) was shown to identify sustained virological response (SVR) with high positive predictive value (93%) and good diagnostic accuracy (81%). The time taken to attain 250 I(eot) ranged from 3 to 11 months in patients with hepatitis C virus (HCV) genotypes 2 or 3 and from 3 to 18 months in those with HCV genotypes 1 or 4. Overall, the duration of therapy would have been 49 months less than that suggested by the most recent algorithms based on a rapid virological response (RVR) at week 4. PMID- 18388886 TI - Mass spectrometry-based proteomics: a useful tool for biomarker discovery? AB - A biomarker is defined as a biological substance (i.e., protein, metabolite, specific post-translational modification) that can be used to detect a disease, measure its progression or the effects of a treatment. Importantly, a biomarker should be readily accessible (i.e., present within body fluids); it must also provide sufficient sensitivity and specificity to accurately distinguish between true positives, false positives, and false negatives. Even more importantly, detection of the biomarker should provide clinical benefits to the patient (i.e., improved survival and/or quality of life). Due to recent technical advances in biomolecular mass spectrometry, a great deal of effort has gone into the discovery of biomarkers at an international level. In this commentary we set forth our views on how mass spectrometry (MS) could be applied to the discovery of elusive biomarkers (Figure 1). PMID- 18388887 TI - Insulin increases tristetraprolin and decreases VEGF gene expression in mouse 3T3 L1 adipocytes. AB - OBJECTIVES: Tristetraprolin (TTP) family proteins (TTP/ZFP36; ZFP36L1, ZFP36L2, ZFP36L3) destabilize adenylate uridylate-rich element-containing mRNAs encoding cytokines, such as tumor necrosis factor (TNF) and vascular endothelial growth factor (VEGF). Little is known about the expression and insulin regulation of TTP and related genes in adipocytes. We analyzed the relative abundance of TTP family mRNAs in 3T3-L1 adipocytes compared to RAW264.7 macrophages and investigated insulin effects on the expression of 43 genes in 3T3-L1 adipocytes. METHODS AND PROCEDURES: Insulin was added to mouse 3T3-L1 adipocytes. Relative abundance of mRNA levels was determined by quantitative real-time PCR. TTP and ZFP36L1 proteins were detected by immunoblotting. RESULTS: Zfp36l1 and Zfp36l2 genes were expressed at eight- to tenfold higher than Ttp in adipocytes. Zfp36l3 mRNA was detected at approximately 1% of Ttp mRNA levels in adipocytes and its low level expression was confirmed in RAW cells. Insulin at 10 and 100 nmol/l increased Ttp mRNA levels by five- to sevenfold, but decreased those of Zfp36l3 by 40% in adipocytes after a 30-min treatment. Immunoblotting showed that insulin induced TTP but did not affect ZFP36L1 protein levels in adipocytes. Insulin decreased mRNA levels of Vegf and a number of other genes in adipocytes. DISCUSSION: Insulin induced Ttp mRNA and protein expression and decreased Vegf mRNA levels in adipocytes. Zfp36l3 mRNA was detected, for the first time, in cells other than mouse placenta and extraembryonic tissues. This study established a basis for the investigation of TTP and VEGF genes in the regulation of obesity and suggested that Vegf mRNA may be a target of TTP in fat cells. PMID- 18388889 TI - Interaction between PPARgamma2 variants and gender on the modulation of body weight. AB - Conflicting results have been reported regarding the effect of the peroxisome proliferator-activated receptor-gamma-2 (PPARgamma2) Pro12Ala polymorphism, (singly or in combination with the silent C1431T polymorphism) on BMI. Gender based dimorphism has been evidenced for genes that affect BMI, but few and conflicting data are available regarding PPARgamma2. We sought to investigate whether the Pro12Ala interacts with gender in modulating BMI in 566 nondiabetic unrelated white subjects (men:women = 211:355, age 36.59 +/- 11.85; BMI 25.36 +/- 4.53). In the whole study population, BMI, fasting glucose and insulin levels, and lipid profile were similar in Ala12 carriers (i.e., XA) and Pro/Pro homozygous subjects. Among the men, but not among the women, X/Ala individuals showed higher BMI (25.9 +/- 3.6 vs. 28.2 +/- 4.9, P = 0.006) and risk of obesity (odds ratio = 2.85, 95% confidence interval = 1.07-7.62). A significant gene gender interaction in modulating BMI was observed (P = 0.039). Among the men, but not among the women, those carrying Ala-T haplotype (i.e., containing both Ala12 and T1431 variants) showed the highest BMI (haplo-score = 3.72, P = 0.0014). Our data indicate that in whites from Italy the PPARgamma2 Pro12Ala polymorphism interacts with gender in modulating BMI, thereby replicating some, but not all, earlier data obtained in different populations. Whether the PPARgamma2-gender interaction is a general phenomenon across different populations, is still an open question, the answer to which requires additional, specifically designed, studies. PMID- 18388888 TI - Weight loss therapy improves pancreatic endocrine function in obese older adults. AB - OBJECTIVE: Obesity and aging increase the risk of type 2 diabetes (T2D). We evaluated whether weight loss therapy improves pancreatic endocrine function and insulin sensitivity in obese older adults. METHODS AND PROCEDURES: Twenty-four obese (BMI: 38 +/- 2 kg/m(2)) older (age: 70 +/- 2 years) adults completed a 6 month randomized, controlled trial. Participants were randomized to diet and exercise (treatment group) or no therapy (control group). beta-Cell function (assessed using the C-peptide minimal model), alpha-cell function (assessed by the glucagon response to an oral glucose load), insulin sensitivity (assessed using the glucose minimal model), and insulin clearance rate were evaluated using a 5-h modified oral glucose tolerance test. RESULTS: Body weight decreased in the treatment group, but did not change in the control group (-9 +/- 1% vs. 0 +/- 1%; P < 0.001). Insulin sensitivity doubled in the treatment group and did not change in the control group (116 +/- 49% vs. -11 +/- 13%; P < 0.05). Even though indices of beta-cell responsivity to glucose did not change (P > 0.05), the disposition index (DI), which adjusts beta-cell insulin response to changes in insulin sensitivity, improved in the treatment group compared with the control group (100 +/- 47% vs. -22 +/- 9%; P < 0.05). The glucagon response decreased in the treatment but not in the control group (-5 +/- 2% vs. 4 +/- 4%; P < 0.05). Insulin secretion rate did not change (P > 0.05), but insulin clearance rate increased (51 +/- 25%; P < 0.05), resulting in lower plasma insulin concentrations. DISCUSSION: Weight loss therapy concomitantly improves beta-cell function, lowers plasma glucagon concentrations, and improves insulin action in obese older adults. These metabolic effects are likely to reduce the risk of developing T2D in this population. PMID- 18388890 TI - Prevalence and characteristics of energy underreporting in African-American girls. AB - OBJECTIVE: To determine the frequency and characteristics of energy intake underreporting in African-American preadolescent girls as part of the Girls health Enrichment Multi-site Studies (GEMS). METHODS AND PROCEDURES: Energy intake was summarized using the Nutrition Data System for Research software and computed as a 3-day average of 24-h dietary recalls. Physical activity was assessed by an accelerometer, basal metabolic rate (BMR) was estimated using the World Health Organization's prediction equation, and underreporting of caloric intake was based on the Goldberg equation. RESULTS: Using a conservative criterion for determining energy underreporting, we classified 54.8% of the girls as underreporters; 45.2% were classified as plausible reporters. Factors related to underreporting included higher BMI (beta = -0.506, P < or = 0.001), older age (beta = -0.159, P = 0.001), greater unhealthy eating behaviors (beta = -0.118, P = 0.025), and higher self-efficacy for diet (beta = -0.098, P = 0.033). DISCUSSION: Underreporting of dietary intake, specifically energy, is common in African-American preadolescent girls and can be partially explained by weight status and psychosocial variables. The extent of dietary underreporting in specific and high-risk populations is largely unknown and could be evaluated by routinely including a report of such an index in future research studies. PMID- 18388891 TI - High-fat diet alters PP2A, TC10, and CIP4 expression in visceral adipose tissue of rats. AB - OBJECTIVE: The aim of this study was to investigate a possible link between high fat diet (HFD)-induced obesity and the expression of protein phosphatase 2A (PP2A) and Cdc42-interacting protein 4 (CIP4) proteins, potential downstream components of the IRS/PI3K/AKT and CAP/Cbl/TC10 pathway, respectively, in the visceral adipose tissue. METHODS AND PROCEDURES: Twenty male Sprague-Dawley rats were randomly divided into two groups and were given either HFD or the normal diet (ND) for 8 weeks. The HFD-induced changes in the expression of the epididymal adipose tissue genes involved in the insulin-signaling pathways were evaluated using real-time reverse-transcription PCR and western blot analysis. RESULTS: The exposure of rats to HFD for 8 weeks resulted in a significant increase in the expression of PP2A at both the transcriptional and translational levels, along with a marked reduction in the levels of phosphorylated AKT and insulin receptor substrate-1 (IRS-1) in the cytosol of visceral adipocytes, compared with the ND rats. Besides, there were significant HFD-induced decreases in the mRNA and protein levels of CIP4 and TC10 in the adipose tissue of rats. DISCUSSION: These data suggest that HFD might have a relevance to insulin resistance by increasing the expression of PP2A, an inhibitor of AKT activity in the phosphatidylinositol 3-kinase (PI3K)/AKT pathway, and also by suppressing the expression of TC10 and CIP4, downstream effectors of the Cbl/CAP/TC10 insulin signaling cascade in the visceral adipose tissue. PMID- 18388892 TI - Mechanism by which BMI influences leisure-time physical activity behavior. AB - OBJECTIVE: The objective of this prospective study was to clarify the mechanism by which BMI influences leisure-time physical activity. This was achieved in accordance with the assumptions underlying the Theory of Planned Behavior (TPB), considered as one of the most useful theories to predict behavior adoption. METHODS AND PROCEDURES: At baseline, a sample of 1,530 respondents completed a short questionnaire to measure intention and perceived behavioral control (PBC), the two proximal determinants of behavior of TPB. Past behavior, sociodemographic variables, and weight and height were also assessed. The dependent variable, leisure-time physical activity was assessed 3 months later. RESULTS: Hierarchical multiple regression analyses revealed that BMI is a direct predictor of future leisure-time physical activity, not mediated by the variables of TPB. Additional hierarchical analyses indicated that BMI was not a moderator of the intention behavior and PBC-behavior relationships. DISCUSSION: The results of this study suggest that high BMI is a significant negative determinant of leisure-time physical activity. This observation reinforces the importance of preventing weight gain as a health promotion strategy for avoiding a sedentary lifestyle. PMID- 18388893 TI - Chromium alleviates glucose intolerance, insulin resistance, and hepatic ER stress in obese mice. AB - OBJECTIVE: Chromium has gained popularity as a nutritional supplement for diabetic patients. This study evaluated the effect of chronic administration of a chromium complex of D-phenylalanine (Cr(D-phe)(3)) on glucose and insulin tolerance in obese mice. The study tested the hypothesis that Cr(D-phe)(3) suppresses endoplasmic reticulum (ER) stress and insulin resistance in these animals. METHODS AND PROCEDURES: C57BL lean and ob/ob obese mice were randomly divided to orally receive vehicle or Cr(D-phe)(3) (3.8 mug of elemental chromium/kg/day) for 6 months. Insulin sensitivity was evaluated by glucose and insulin tolerance tests. Protein levels of phosphorylated pancreatic ER kinase (PERK), alpha subunit of translation initiation factor 2 (eIF2alpha) and inositol requiring enzyme-1 (IRE-1), p-c-Jun, and insulin receptor substrate-1 (IRS-1) phosphoserine-307 were assessed by western blotting. In vitro ER stress was induced by treating cultured muscle cells with thapsigargin in the presence or absence of Cr(D-phe)(3). RESULTS: ob/ob mice showed poor glucose and insulin tolerance compared to the lean controls, which was attenuated by Cr(D-phe)(3). Markers of insulin resistance (phospho-c-Jun and IRS-1 phosphoserine) and ER stress (p-PERK, p-IRE-1, p-eIF2alpha), which were elevated in ob/ob mice, were attenuated following Cr(D-phe)(3) treatment. Chromium treatment was also associated with a reduction in liver triglyceride levels and lipid accumulation. In cultured myotubes, Cr(D-phe)(3) attenuated ER stress induced by thapsigargin. DISCUSSION: Oral Cr(D-phe)(3) treatment reduces glucose intolerance, insulin resistance, and hepatic ER stress in obese, insulin-resistant mice. PMID- 18388894 TI - Association between BMI and metabolic syndrome and adenomatous colonic polyps in Korean men. AB - Obesity and insulin resistance are associated with the risk of colon cancer. Adenomatous colonic polyps are precancerous lesions of colon cancer. We investigated whether BMI and the metabolic syndrome are associated with the presence of adenomatous colonic polyps in Korean men. Anthropometric measurements, metabolic risk factors, and colonoscopic pathologic findings were assessed in 1,898 men who underwent routine colonoscopy at the Health Promotion Center of Asan Medical Center in 2005. The modified National Cholesterol Education Program Adult Treatment Panel III (NCEP-ATP III) and International Diabetes Federation (IDF) criteria were used for the definition of the metabolic syndrome. Multiple logistic regression analysis was used to evaluate the association between BMI and the metabolic syndrome and adenomatous polyps. Compared with men in the 1st quintile of the BMI, the adjusted odds ratio (OR) and 95% confidence interval (CI) for adenomatous polyps in men in the 2nd, 3rd, 4th, and 5th quintiles of the BMI were 1.55 (1.10-2.19), 1.57 (1.10-2.24), 1.94 (1.34-2.81), and 1.99 (1.31-3.01), respectively (P for trend <0.0001). Men with triglycerides (TGs) > or = 150 mg/dl were significantly more likely to have adenomatous polyps than were men with TG <150 mg/dl (OR 1.29; 95% CI 1.03-1.62). As a function of the number of metabolic risk factors, the ORs for adenomatous polyps were 1.41 (1.03-1.93), 1.52 (1.08-2.12), 1.46 (1.01-2.12), and 1.77 (1.08 2.90) for 1, 2, 3, and > or = 4 risk factors, respectively (P for trend <0.05). Adenomatous colonic polyps were significantly associated with increased BMI levels. Subjects with even one component of the metabolic syndrome had a significantly higher risk for developing adenomatous polyps compared to those subjects without any component in Korean men. PMID- 18388895 TI - Motor skill performance and physical activity in preschool children. AB - Children with better-developed motor skills may find it easier to be active and engage in more physical activity (PA) than those with less-developed motor skills. The purpose of this study was to examine the relationship between motor skill performance and PA in preschool children. Participants were 80 three- and 118 four-year-old children. The Children's Activity and Movement in Preschool Study (CHAMPS) Motor Skill Protocol was used to assess process characteristics of six locomotor and six object control skills; scores were categorized as locomotor, object control, and total. The actigraph accelerometer was used to measure PA; data were expressed as percent of time spent in sedentary, light, moderate-to-vigorous PA (MVPA), and vigorous PA (VPA). Children in the highest tertile for total score spent significantly more time in MVPA (13.4% vs. 12.8% vs. 11.4%) and VPA (5% vs. 4.6% vs. 3.8%) than children in middle and lowest tertiles. Children in the highest tertile of locomotor scores spent significantly less time in sedentary activity than children in other tertiles and significantly more time in MVPA (13.4% vs. 11.6%) and VPA (4.9% vs. 3.8%) than children in the lowest tertile. There were no differences among tertiles for object control scores. Children with poorer motor skill performance were less active than children with better-developed motor skills. This relationship between motor skill performance and PA could be important to the health of children, particularly in obesity prevention. Clinicians should work with parents to monitor motor skills and to encourage children to engage in activities that promote motor skill performance. PMID- 18388896 TI - Elevated soluble lectin-like oxidized LDL receptor-1 (sLOX-1) levels in obese postmenopausal women. AB - We investigated the association between soluble lectin-like oxidized low-density lipoprotein receptor-1 (sLOX-1) levels and obesity in older women. Fifty-one postmenopausal women (10 lean, 22 overweight, and 19 obese) were included in this small retrospective analysis. Plasma sLOX-1 levels were measured using a chemiluminescent enzyme-linked immunoassay. Plasma levels of sLOX-1 were significantly higher in obese women (55.33 +/- 4.49 pg/ml) compared to lean (30.91 +/- 6.19 pg/ml, P = 0.002) and overweight women (38.31 +/- 4.18 pg/ml, P = 0.017). Plasma sLOX-1 levels were positively associated with body weight, BMI, total body fat, and trunk fat. The relationship between sLOX-1 and BMI was attenuated after adjustment for age, hormone replacement therapy, and body fat. In conclusion, obese women have higher sLOX-1 levels, which may reflect increased LOX-1 expression in adipose tissue. PMID- 18388897 TI - Short-term training effects on diastolic function in obese persons with the metabolic syndrome. AB - The aim of this study was to determine the effects of a short-term high-intensity exercise program on diastolic function and glucose tolerance in obese individuals with and without metabolic syndrome (MetSyn). Obese men and women (BMI > 30 kg/m(2); 39-60 years) with and without the MetSyn (MetSyn 13; non-MetSyn 18) underwent exercise training consisting of 10 consecutive days of treadmill walking for 1 h/day at 70-75% of peak aerobic capacity. Subjects performed pre- and post-training testing for aerobic capacity, glucose tolerance (2-h meal test), and standard echocardiography. Aerobic capacity improved for both groups (non-MetSyn 24.0 +/- 1.6 ml/kg/min vs. 25.1 +/- 1.5 ml/kg/min; MetSyn 25.2 +/- 1.8 ml/kg/min vs. 26.2 +/- 1.7 ml/kg/min, P < 0.05). Glucose area under the curve (AUC) improved in the MetSyn group (1,017 +/- 58 pmol/l/min vs. 883 +/- 75 pmol/l/min, P < 0.05) with no change for the non-MetSyn group (685 +/- 54 pmol/l/min vs. 695 +/- 70 pmol/l/min). Isovolumic relaxation time (IVRT) improved in the MetSyn group (97 +/- 6 ms vs. 80 +/- 5 ms, P < 0.05), and remained normal in the non-MetSyn group (82 +/- 6 ms vs. 86 +/- 5 ms). No changes in other diastolic parameters were observed. The overall reduction in IVRT was correlated with a decrease in diastolic blood pressure (DBP) (r = 0.45, P < 0.05), but not with changes in glucose tolerance. Body weight did not change with training in either group. A 10-day high-intensity exercise program improved diastolic function and glucose tolerance in the group with MetSyn. The reduction in IVRT in MetSyn was associated with a fall in blood pressure. These data suggest that it may be possible to reverse early parameters of diastolic dysfunction in MetSyn with a high-intensity exercise program. PMID- 18388898 TI - INS VNTR is not associated with childhood obesity in 1,023 families: a family based study. AB - Previous studies have described genetic associations of the insulin gene variable number tandem repeat (INS VNTR) variant with childhood obesity and associated phenotypes. We aimed to assess the contribution of INS VNTR genotypes to childhood obesity and variance of insulin resistance, insulin secretion, and birth weight using family-based design. Participants were either French or German whites. We used transmission disequilibrium tests (TDTs) for assessing binary traits and quantitative pedigree disequilibrium tests for assessing continuous traits. In contrast to previous findings, we did not observe any familial association with childhood obesity (T = 50%, P = 0.77) in the 1,023 families tested. In French obese children, INS VNTR did not associate with fasting insulin levels (P = 0.23) and class I allele showed only borderline association with increased insulin secretion index at 30 min (P = 0.03). INS VNTR did not associate with birth weight in obese children (P = 0.98) and TDT analyses in 350 French families with history of low birth weight (LBW) showed no association with this condition (P = 0.92). In summary, our study, the largest performed so far, does not support the previously reported associations between INS VNTR and childhood obesity, insulin resistance, or birth weight, and does not suggest any major role for this variant in modulating these traits. PMID- 18388899 TI - Trends in overweight and obesity in Chinese adults: between 1991 and 1999-2000. AB - OBJECTIVE: The aim of this study was to evaluate trends in BMI and the prevalence of overweight (BMI > or = 25 kg/m(2)) and obesity (BMI > or = 30 kg/m(2)) between 1991 and 1999-2000 among Chinese adults. METHODS AND PROCEDURES: In this study, two population-based samples of Chinese adults aged between 45 and 79 years (n = 7,858 during each period), and comparable in the distributions of age, gender, degree of urbanization, and region (North/South) were used. Height and weight were measured using identical procedures at each period, and BMI was calculated as weight (in kilogram) divided by height (in square meter). RESULTS: From 1991 to 1999-2000, the mean BMI increased from 21.8 to 23.4 kg/m(2) among men and from 21.8 to 23.5 kg/m(2) among women (each P < 0.001). Among men, the prevalence of overweight and obesity increased from 9.6 and 0.6%, respectively, in 1991 to 20.0 and 3.0%, respectively, in 1999-2000 (each P < 0.001). Among women, the prevalence of overweight and obesity increased from 14.5 and 1.8%, respectively, in 1991 to 26.5 and 5.2%, respectively, in 1999-2000 (each P < 0.001). The prevalence of overweight and obesity increased in all age groups, in rural and urban areas, and in North and South China, with greater relative increases in obesity among older age groups, South China, and rural areas (P interaction < 0.05). DISCUSSION: Overweight and obesity increased tremendously during the 1990s in China. These data underscore the need for national programs in weight maintenance and reduction, to prevent obesity-related outcomes in China. PMID- 18388901 TI - Impact of social support intensity on walking in the severely obese: a randomized clinical trial. AB - OBJECTIVE: There are few established methods for promoting physical activity (PA) in the severely obese. Because social support is a potential method for promoting PA, we compared mean steps/day during 18 weeks in severely obese outpatients receiving either standard support (SS) or added support (AS). METHODS AND PROCEDURES: Eighty severely obese outpatients from an obesity clinic were invited; 66 provided written consent, 55 were randomized, and 42 were included in final analyses (9 men, 33 women; age 44.4 +/- 13.1 years; BMI 41.9 +/- 5.5 kg/m(2)). All participants received a pedometer and a walking promotion booklet. In addition to SS, the AS group received ten 2-h group counseling sessions aimed at increasing weekly accumulated steps, every second week during the study. Each participant was asked to complete a 7-day walking diary every second week (10 observations). RESULTS: Baseline steps/day was 6,912 for the AS group and 5,311 for the SS group (P = 0.023). Data at 18 weeks showed that the AS group recorded 10,136 steps/day and the SS group 6,118 steps/day (P = 0.024). There was no allocation x time interaction (P = 0.46). During the follow-up period as a whole, the AS group recorded 1,794 more steps/day than the SS group (P = 0.0074). DISCUSSION: The AS group recorded more steps/day than the SS group, reaching a mean level of approximately 10,000 steps/day. However, the nonsignificant interaction between allocation x time suggests that this difference was present already at baseline and did not increase during follow-up. PMID- 18388900 TI - Depot-specific regulation of the conversion of cortisone to cortisol in human adipose tissue. AB - OBJECTIVE: Our main objective was to compare the regulation of cortisol production within omental (Om) and abdominal subcutaneous (Abd sc) human adipose tissue. METHODS AND PROCEDURES: Om and Abd sc adipose tissue were obtained at surgery from subjects with a wide range of BMI. Hydroxysteroid dehydrogenase (HSD) activity ((3)H-cortisone and (3)H-cortisol interconversion) and expression were measured before and after organ culture with insulin and/or dexamethasone. RESULTS: Type 1 HSD (HSD1) mRNA and reductase activity were mainly expressed within adipocytes and tightly correlated with adipocyte size within both depots. There was no depot difference in HSD1 expression or reductase activity, while cortisol inactivation and HSD2 mRNA expression (expressed in stromal cells) were higher in Om suggesting higher cortisol turnover in this depot. Culture with insulin decreased HSD reductase activity in both depots. Culture with dexamethasone plus insulin compared to insulin alone increased HSD reductase activity only in the Om depot. This depot-specific increase in reductase activity could not be explained by an alteration in HSD1 mRNA or protein, which was paradoxically decreased. However, in Om only, hexose-6-phosphate dehydrogenase (H6PDH) mRNA levels were increased by culture with dexamethasone plus insulin compared to insulin alone, suggesting that higher nicotinamide adenine dinucleotide phosphate-oxidase (NADPH) production within the endoplasmic reticulum (ER) contributed to the higher HSD reductase activity. DISCUSSION: We conclude that in the presence of insulin, glucocorticoids cause a depot-specific increase in the activation of cortisone within Om adipose tissue, and that this mechanism may contribute to adipocyte hypertrophy and visceral obesity. PMID- 18388902 TI - Eating meals irregularly: a novel environmental risk factor for the metabolic syndrome. AB - BACKGROUND: Skipping meals is a common practice in our current society; however, it is not clear whether eating meals regularly is associated with the metabolic syndrome. OBJECTIVE: Our aim was to assess the association of eating meals regularly with parameters of the metabolic syndrome and insulin resistance in a representative population-based cohort of 60-year-old men and women. METHODS AND PROCEDURES: A population-based cross-sectional study of 3,607 individuals (1,686 men and 1,921 women), aged 60 years, was conducted in Stockholm County, Sweden. Medical history, socioeconomic factors, and lifestyle data were collected by a questionnaire and a medical examination, which included laboratory tests. RESULTS: Of the subjects who were regular eaters, 20% fulfilled the criteria for the metabolic syndrome vs. 27% of subjects who were irregular eaters (P < 0.0001). The adjusted odds ratio (OR) for having the greatest number of components of the metabolic syndrome in subjects who were regular eaters was 0.27 (95% confidence interval (CI), 0.13-0.54) using subjects who did not fulfill any criteria for the metabolic syndrome as a reference group. Eating meals regularly was also inversely related to insulin resistance (OR, 0.68 (95% CI, 0.48-0.97)) and to gamma-glutamyl transferase (OR, 0.52 (95% CI, 0.33-83)) after full adjustment. DISCUSSION: Eating meals regularly is inversely associated to the metabolic syndrome, insulin resistance and (high) serum concentrations of gamma glutamyl transferase. These findings suggest that eating meals irregularly may be part of several potential environmental risk factors that are associated with the metabolic syndrome and may have future implications in giving dietary advice to prevent and/or treat the syndrome. PMID- 18388903 TI - Obesity-related changes in high-density lipoprotein metabolism. AB - Obesity is associated with a 3-or-more-fold increase in the risk of fatal and nonfatal myocardial infarction (1,2,3,4,5,6). The American Heart Association has reclassified obesity as a major, modifiable risk factor for coronary heart disease (7). The increased prevalence of premature coronary heart disease in obesity is attributed to multiple factors (8,9,10). A principal contributor to this serious morbidity is the alterations in plasma lipid and lipoprotein levels. The dyslipidemia of obesity is commonly manifested as high plasma triglyceride levels, low high-density lipoprotein cholesterol (HDLc), and normal low-density lipoprotein cholesterol (LDLc) with preponderance of small dense LDL particles (7,8,9,10). However, there is a considerable heterogeneity of plasma lipid profile in overweight and obese people. The precise cause of this heterogeneity is not entirely clear but has been partly attributed to the degree of visceral adiposity and insulin resistance. The emergence of glucose intolerance or a genetic predisposition to familial combined hyperlipidemia will further modify the plasma lipid phenotype in obese people (11,12,13,14,15). PMID- 18388904 TI - Cleaning up the EPA's act. PMID- 18388905 TI - INO80 meets a fork in the road. PMID- 18388906 TI - Think you know how miRNAs work? Think again. PMID- 18388907 TI - No SIRT6. PMID- 18388909 TI - Lentiviral vector-mediated SERCA2 gene transfer protects against heart failure and left ventricular remodeling after myocardial infarction in rats. AB - Reduced expression of the SERCA2 gene impairs the calcium-handling and contractile functions of the heart. We developed an SERCA2 gene transfer system using lentiviral vectors, and examined the long-term effect of SERCA2 gene transfer in the rat ischemic heart failure model. A lentiviral vector containing the SERCA2 gene was infused into a rat heart by hypothermic intracoronary delivery 2 weeks after myocardial infarction (MI). The transduction efficiency was approximately 40%. Six months after transduction, echocardiogram and pressure volume measurements revealed that the SERCA2 gene transfer had significantly protected against left ventricular (LV) dilation, and had improved systolic and diastolic function, resulting in reduction in mortality rates. The brain natriuretic peptide mRNA level showed a significantly decrease and the phosphorylation level of serine residue of phospholamban (PLN) showed an increase in the Lenti-SERCA2-transduced heart. Further, DNA microarray analysis disclosed that SERCA2 gene transfer had increased cardioprotective gene expression and lowered the expression of genes that are known to exacerbate heart failure. The SERCA2 gene was successfully integrated into the host heart, induced favorable molecular remodeling, prevented LV geometrical remodeling, and improved the survival rate. These results suggest that a strategy to compensate for reduced SERCA2 gene expression by lentiviral vectors serves as a positive inotropic, lucitropic, and cardioprotective therapy for post-MI heart failure. PMID- 18388910 TI - Delivery of AAV-IGF-1 to the CNS extends survival in ALS mice through modification of aberrant glial cell activity. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of the motor system. Recent work in rodent models of ALS has shown that insulin-like growth factor-1 (IGF-1) slows disease progression when delivered at disease onset. However, IGF-1's mechanism of action along the neuromuscular axis remains unclear. In this study, symptomatic ALS mice received IGF-1 through stereotaxic injection of an IGF-1-expressing viral vector to the deep cerebellar nuclei (DCN), a region of the cerebellum with extensive brain stem and spinal cord connections. We found that delivery of IGF-1 to the central nervous system (CNS) reduced ALS neuropathology, improved muscle strength, and significantly extended life span in ALS mice. To explore the mechanism of action of IGF-1, we used a newly developed in vitro model of ALS. We demonstrate that IGF-1 is potently neuroprotective and attenuates glial cell-mediated release of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO). Our results show that delivering IGF-1 to the CNS is sufficient to delay disease progression in a mouse model of familial ALS and demonstrate for the first time that IGF-1 attenuates the pathological activity of non-neuronal cells that contribute to disease progression. Our findings highlight an innovative approach for delivering IGF-1 to the CNS. PMID- 18388911 TI - Heterologous boosting of recombinant adenoviral prime immunization with a novel vesicular stomatitis virus-vectored tuberculosis vaccine. AB - Pulmonary tuberculosis (TB) remains a serious health problem worldwide. Effective vaccination strategies are needed. We report the development of a novel TB vaccine using vesicular stomatitis virus (VSV) as a viral vector system to express Ag85A. VSVAg85A was shown to be immunogenic when given to mice by either an intranasal or an intramuscular (i.m.) route. Although distinct T-cell profiles resulted from both routes of immunization, only intranasal delivery generated a mucosal T-cell response that was protective upon pulmonary Mycobacterium tuberculosis (M.tb) challenge. While this protection manifested at an early time point after immunization, it was not sustained. The potential of VSVAg85A to be used as a mucosal booster for parenteral priming by an adenoviral TB vaccine expressing Ag85A (AdAg85A) was investigated. VSVAg85A immunization markedly boosted antigen-specific T-cell responses in the airway lumen while also augmenting immune activation in the systemic compartment, after AdAg85A priming. This translated into significantly better protective efficacy against pulmonary challenge with M.tb than either vaccine used alone. Our study therefore suggests that VSV as a vector system is a promising candidate to be used in a heterologous viral prime-boost immunization regimen against intracellular bacterial infection. PMID- 18388912 TI - Trichostatin A and oncolytic HSV combination therapy shows enhanced antitumoral and antiangiogenic effects. AB - Oncolytic herpes simplex viruses (HSVs) possess direct oncolytic and antiangiogenic activities and are promising anticancer agents, but their efficacy, when used as single agents, leaves room for improvement. We investigated whether combination therapy of HSV with histone deacetylase inhibitor trichostatin A (TSA), an agent that also targets cancer cells and tumor vasculature, would result in enhanced efficacy. In vitro, TSA and G47Delta showed strong synergy of action against proliferating endothelial cells, varying degrees of synergistic action against most cancer cell lines, but no effect in quiescent, normal endothelial and prostate epithelial cells. Synergy is dependent on viral replication; however, it is not dependent on the dosing sequence of TSA and G47Delta, viral genetic alterations, infectivity, or replication kinetics of G47Delta. Using an isogenic cell system, we found that a high level of cellular cyclin D1 is also critically important for the interaction. Normal cells with low cyclin D1 levels were not subjected to toxicity by either agent. In tumor cells and proliferating endothelial cells, the combination treatment enhanced the inhibition of cyclin D1 and vascular endothelial growth factor (VEGF). Concurrent systemic TSA and intratumoral G47Delta administration resulted in enhanced antiangiogenesis and enhanced antitumoral efficacy in animal models. Therefore, combination treatment with TSA and oncolytic HSV provides a novel approach to cancer therapy. PMID- 18388913 TI - Effect of late-stage therapy on disease progression in AAV-mediated rescue of photoreceptor cells in the retinoschisin-deficient mouse. AB - Proof-of-concept for a successful adeno-associated virus serotype 5 (AAV5) mediated gene therapy in X-linked juvenile retinoschisis (XLRS) has been demonstrated in an established mouse model for this condition. The initial studies concentrated on early time-points of treatment. In this study, we aimed to explore the consequences of single subretinal injections administered at various stages of more advanced disease. By electroretinogram (ERG), functional improvement in treated versus untreated eyes is found to be significant in retinoschisin-deficient mice injected at the time-points of 15 days (P15), 1 month (PM1), and 2 months (PM2) after birth. In mice treated at 7 months after birth (PM7), an age previously shown to exhibit advanced retinal disease, ERG responses reveal no beneficial effects of vector treatment. Generally, functional rescue is paralleled by sustained retinoschisin expression and significant photoreceptor survival relative to untreated eyes. Quantitative measures of photoreceptors and peanut agglutinin-labeled ribbon synapses demonstrate rescue effects even in mice injected as late as PM7. Taken together, AAV5-mediated gene replacement is beneficial in slowing disease progression in murine XLRS. In addition, we show the effectiveness of rescue efforts even if treatment is delayed until advanced signs of disease have developed. Human XLRS patients might benefit from these findings, which suggest that the effectiveness of treatment appears not to be restricted to the early stages of the disease, and that treatment may prove to be valuable even when administered at more advanced stages. PMID- 18388914 TI - Extended transgene expression from a nonintegrating adenoviral vector containing retroviral elements. AB - We studied the effects of specific retroviral elements in a first-generation serotype 5 adenoviral (Ad5) vector, AdLTR(2)EF1alpha-hEPO. This vector contains 858 base pair (bp) [251-bp envelope sequence plus 607-bp long-terminal repeat (LTR)] from Moloney murine leukemia virus (MoMLV), upstream of the human elongation factor-1alpha (EF1alpha) promoter and human erythropoietin (hEPO) cDNA, with the LTR sequence downstream of the polyadenylation signal. We compared expression of AdLTR(2)EF1alpha-hEPO with corresponding expressions of two conventional Ad5 vectors, AdEF1alpha-hEPO and AdCMV-hEPO, in vivo in submandibular glands in rats. Both the conventional vectors yielded low serum hEPO levels by day 7, and little change in hematocrits. In contrast, after receiving AdLTR(2)EF1alpha-hEPO, the rats showed elevated hEPO levels and hematocrits for 1-3 months. In vitro studies showed that the integration efficiencies of all the vectors were similar (approximately 10(-3)). Approximately 0.1% of the vector genomes were present 1 year after delivery in the case of each of the three vectors, primarily as intact linear double-strand DNA. The unique results seen with AdLTR(2)EF1alpha-hEPO are partly because of LTR enhancer activity. However, other cis-acting activity, which is not immunomodulatory but nevertheless influences promoter methylation, appears to be involved. A vector such as AdLTR(2)EF1alpha-hEPO may prove useful in clinical applications in which extended, but not "permanent," transgene expression is desirable. PMID- 18388915 TI - Use of a U16 snoRNA-containing ribozyme library to identify ribozyme targets in HIV-1. AB - Hammerhead ribozymes have been shown to silence human immunodeficiency virus-1 (HIV-1) gene expression by site-specific cleavage of viral mRNA. The two major factors that determine whether ribozymes will be effective for post transcriptional gene silencing are colocalization of the ribozyme and the target RNAs, and the choice of an appropriate target site on the mRNA. An effective screening strategy for potential targets on the viral genome is the use of ribozyme libraries in cell culture. Capitalizing on previous findings that HIV-1 and ribozymes can be colocalized in the nucleolus, we created a novel hammerhead ribozyme library by inserting hammerhead ribozymes with fully randomized stems 1 and 2 into the body of the U16 small nucleolar RNA (snoRNA). Following three rounds of cotransfection with an HIV-1 proviral DNA harboring the herpes simplex virus thymidine kinase (HSV-TK) gene, we selected for gancyclovir-resistant cells and identified a ribozyme sequence that could potentially target both the U5 and gag genes of HIV-1 regions on the HIV-1 genome through partial homologies with these targets. When the ribozymes were converted to full complementarity with the targets, they provided potent inhibition of HIV-1 replication in cell culture. These results provide a novel approach for identifying ribozyme targets in HIV-1. PMID- 18388916 TI - Efficient oncogene silencing and metastasis inhibition via systemic delivery of siRNA. AB - The selective delivery of small interfering RNA (siRNA) to metastatic tumors remains a challenging task. We have developed a nanoparticle (NP) formulation composed of siRNA, a carrier DNA, a polycationic peptide, and cationic liposomes. The NP was obtained by a self-assembling process, followed by surface modification with a polyethylene glycol (PEG)-conjugated ligand, anisamide. The NP was PEGylated and a ligand was presented to target sigma receptor-expressing murine melanoma cells, B16F10. The lung metastasis model was established by intravenous (i.v.) injection of the B16F10 cells into C57BL/6 mice. A mixture of siRNA against MDM2, c-myc, and vascular endothelial growth factor (VEGF) co formulated in the targeted NP caused simultaneous silencing of each of the oncogenes in the metastatic nodules. Two consecutive i.v. injections of siRNA in the targeted NP significantly reduced the lung metastasis (approximately 70-80%) at a relatively low dose (0.45 mg/kg), whereas free siRNA and the nontargeted NP showed little effect. This targeted NP formulation significantly prolonged the mean survival time of the animals by 30% as compared to the untreated controls. At the therapeutic dose, the targeted NP showed little local and systemic immunotoxicity and did not decrease the body weight or damage the major organs. PMID- 18388917 TI - AAV vector-mediated RNAi of mutant huntingtin expression is neuroprotective in a novel genetic rat model of Huntington's disease. AB - We report the characterization of a new rapid-onset model of Huntington's disease (HD) generated by adeno-associated virus (AAV) vector-mediated gene transfer of N terminal huntingtin (htt) constructs into the rat striatum. Expression of exon 1 of mutant htt containing 70 CAG repeats rapidly led to neuropathological features associated with HD. In addition, we report novel data relating to neuronal transduction of AAV vectors that modulated the phenotype observed in this model. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that AAV vector-mediated expression in the striatum increased by >100-fold as compared to the endogenous htt level. Moreover, AAV vectors exhibited nonuniform transduction patterns in striatal neuronal populations, as well as axonal transport leading to transduction and neuronal cell death in the globus pallidus and substantia nigra (SN). These findings may inform future studies that utilize AAV vectors for neurodegenerative disease modeling. Further, RNA interference (RNAi) of mutant htt expression mediated by virus vector delivery of short hairpin RNAs (shRNAs) ameliorates early-stage disease phenotypes in transgenic mouse models of HD. However, it has not been reported whether shRNA-mediated knockdown of mutant htt expression is neuroprotective. AAV-shRNA was shown to mediate a dramatic knockdown of HD70 expression, preventing striatal neurodegeneration and concomitant motor behavioral impairment. These results provide further support for the use of AAV vector-mediated RNAi as a therapeutic strategy for HD. PMID- 18388918 TI - Efficacy and safety of the oncolytic herpes simplex virus rRp450 alone and combined with cyclophosphamide. AB - Oncolytic herpes simplex virus (oHSV) mutants are under development as anticancer therapeutics. One such vector, rRp450, is ICP6-deleted and expresses a prodrug enzyme for cyclophosphamide (CPA) (rat CYP2B1). Little is known about rRp450's toxicity profile, especially in combination with CPA. We tested rRp450/CPA for antitumor efficacy in an aggressive human xenograft sarcoma model, measured virus production in primary cells, and tested rRp450/CPA for safety in immunocompetent mice. CPA enhanced the antitumor efficacy of rRp450. Relative to wild-type HSV-1, rRp450 replication was attenuated approximately 10,000-fold in human primary hepatocytes, differentiated primary foreskin keratinocytes, and primary Schwann cells. In vivo, intravenous and intracranial (IC) rRp450 injection at the strength of 10(8) plaque-forming units (pfu) alone or followed 24 hours later by intraperitoneal (IP) CPA was well tolerated and had no significant effect clinically on blood counts or chemistries. By contrast, intravenous KOS was found to be uniformly neurotoxic at 10(5) and fatal at 10(6) pfu, and IC virus was fatal in most mice at 10(4) pfu. Low levels of virus DNA were detected in some organs following intravenous and IC virus injection, but were not significantly altered by CPA. HSV replication was not detected in reactivation studies of isolated organs. Our findings suggest rRp450/CPA is safe and warrants further study as a potential combination in anticancer therapeutics. PMID- 18388919 TI - Systemic osteoprotegerin gene therapy restores tumor-induced bone loss in a therapeutic model of breast cancer bone metastasis. AB - Enhanced production of receptor activator of nuclear factor-kappaB ligand (RANKL) and its binding to RANK on the osteoclasts have been associated with osteolysis in breast cancer bone metastasis. Osteoprotegerin (OPG) is a decoy receptor that prevents RANKL-RANK interaction. This study determined the effects of sustained expression of OPG using a recombinant adeno-associated viral (rAAV) vector in mouse model of osteolytic breast cancer. Bone metastasis was established by intracardiac injection of the human breast cancer cell line MDA-MB-435. Following this, mice were administered a one-time intramuscular injection of rAAV encoding either OPG.Fc (OPG) or green fluorescent protein (GFP). Mice were killed 1 month later and the effects of therapy on tumor growth and bone remodeling were evaluated. Bioluminescence imaging showed significant reduction of tumor growth in bone of OPG.Fc-treated mice. Micro-computed tomography (microCT) analysis and histomorphometry of the tibia indicated significant protection of trabecular and cortical bones after OPG.Fc therapy. Despite the prevention of bone loss and tumor growth in bone, OPG.Fc therapy failed to provide long-term survival. OPG.Fc treated mice developed more bone than age-matched normal mice, indicating a requirement for regulated transgene expression. Results of this study indicate the potential of rAAV-OPG therapy for reducing morbidity and mortality in breast cancer patients with osteolytic bone damage. PMID- 18388921 TI - Improvement of migratory defects in a murine model of Wiskott-Aldrich syndrome gene therapy. AB - Wiskott-Aldrich syndrome (WAS) is an X-linked hematological disease characterized by immunodeficiency, eczema, and thrombocytopaenia, and shows promise for treatment with hematopoietic stem cell gene therapy. The immunopathology of WAS is attributable at least in part to defects of cell migration and localization as a result of chemotactic, adhesive, and chemokinetic defects. Whereas previous studies using either gammaretroviral or lentiviral vectors have demonstrated variable correction of T-cell proliferation and dendritic cell (DC) cytoarchitecture, we have used a lentiviral vector expressing an eGFP-WASp fusion protein to test the potential for restoration of cell migratory defects. Multilineage expression of the fusion transgene was present for up to 10 months after primary engraftment, and also in secondary recipients analyzed after a further 9 months. Transduced bone marrow-derived dendritic cells (BMDCs) demonstrated recovery of podosome numbers and turnover, while B cells, BMDCs, and Langerhans cells (LCs) exhibited enhanced chemotactic responses to specific stimuli. As an indication of functionality in vivo, splenic marginal zone B cells and a cutaneous contact hypersensitivity (CHS) response to dinitrofluorobenzene (DNFB) were both partially restored. These proof of principle experiments demonstrate that WAS protein (WASp) transgene expression can be successfully maintained long term in primary and secondary recipients, and that it is associated with a significant repair of migratory defects. PMID- 18388920 TI - Therapeutic molecular targeting of 15-lipoxygenase-1 in colon cancer. AB - Molecular targeting for apoptosis induction is being developed for better treatment of cancer. Downregulation of 15-lipoxygenase-1 (15-LOX-1) is linked to colorectal tumorigenesis. Re-expression of 15-LOX-1 in cancer cells by pharmaceutical agents induces apoptosis. Antitumorigenic agents can also induce apoptosis via other molecular targets. Whether restoring 15-LOX-1 expression in cancer cells is therapeutically sufficient to inhibit colonic tumorigenesis remains unknown. We tested this question using an adenoviral delivery system to express 15-LOX-1 in in vitro and in vivo models of colon cancer. We found that (i) the adenoviral vector 5/3 fiber modification enhanced 15-LOX-1 gene transduction in various colorectal cancer cell lines, (ii) the adenoviral vector delivery restored 15-LOX-1 expression and enzymatic activity to therapeutic levels in colon cancer cell lines, and (iii) 15-LOX-1 expression downregulated the expression of the antiapoptotic proteins X-linked inhibitor of apoptosis protein (XIAP) and BcL-XL, activated caspase-3, triggered apoptosis, and inhibited cancer cell survival in vitro and the growth of colon cancer xenografts in vivo. Thus, selective molecular targeting of 15-LOX-1 expression is sufficient to re-establish apoptosis in colon cancer cells and inhibit tumorigenesis. These data provide the rationale for further development of therapeutic strategies to target 15-LOX-1 molecularly for treating colonic tumorigenesis. PMID- 18388922 TI - Subretinal delivery of recombinant AAV serotype 8 vector in dogs results in gene transfer to neurons in the brain. AB - Recombinant adeno-associated virus (rAAV) vectors are among the most efficient gene delivery vehicles for gene transfer to the retina. This study evaluates the behavior of the rAAV8 serotype vector with regard to intraocular delivery in rats and dogs. Subretinal delivery of an AAV2/8.gfp vector results in efficient gene transfer in the retinal pigment epithelium (RPE), the photoreceptors and, surprisingly, in the cells of the inner nuclear layer as well as in ganglion cells. Most importantly, in dogs, gene transfer also occurred distal to the injection site in neurons of the lateral geniculate nucleus of the brain. Because green fluorescent protein (GFP) was detected along the visual pathway within the brain, we analyzed total DNA extracted from various brain slices using PCR. Vector sequences were detected in many parts of the brain, but chiefly in the contralateral hemisphere. PMID- 18388923 TI - Administering plasmid DNA encoding tumor vessel-anchored IFN-alpha for localizing gene product within or into tumors. AB - Tumor-targeted gene delivery has been intensively studied in the field of gene therapy, but no attention has been given to targeting the therapeutic gene products, which are transcribed and translated from the injected genes, into tumors. Targeting immune stimulatory gene products into tumors is the key to triggering tumor-specific CD8(+) T-cell responses and reducing systemic toxicity. To target the gene products generated from the injected genes into tumors, genes encoding the tumor-targeted fusion gene product were generated and administered locally and systemically via electroporation. As anticipated, administration of a therapeutic gene encoding IFN-alpha and the tumor vessel-targeted peptide CDGRC fusion gene product minimizes the leakage of immunostimulatory cytokine from tumors into the blood circulation, increases the infiltration of CD8(+) T cells into tumors, induces a high magnitude of cytotoxic T-cell lysis (CTL) activity, and reduces tumor vessel density. As a result, tumor growth was more significantly inhibited by administering the IFN-alpha-CDGRC gene than by administering the wild-type IFN-alpha gene. The same result was obtained with the systemic administration of the tumor-targeted IFN-alpha gene. This gene product based tumor-targeted gene therapy approach could complement any other tumor targeted gene delivery method for improving tumor-targeting efficiency. PMID- 18388924 TI - Reduced pathology and improved behavioral performance in Alzheimer's disease mice vaccinated with HSV amplicons expressing amyloid-beta and interleukin-4. AB - Immunotherapeutics designed to dissolve existing amyloid plaques or to interrupt amyloid-beta (Abeta) accumulation may be feasible for treatment and/or prevention of Alzheimer's disease (AD). "Shaping" the immune responses elicited against Abeta is requisite toward generating an efficacious and safe outcome; this can be achieved by minimizing the possibility of deleterious inflammatory reactions in the brain as observed in clinical testing of Abeta peptide/adjuvant-based modalities. Herpes simplex virus (HSV)-based amplicons can coexpress multiple antigens and/or immunomodulatory genes due to their large genetic size capacity, thereby facilitating antigen-specific immune response shaping. We have constructed an amplicon (HSV(IE)Abeta(CMV)IL-4) that co-delivers Abeta(1-42) with interleukin-4 (IL-4), a cytokine that promotes the generation of Th2-like T-cell responses, which are favored in the setting of AD immunotherapy. Triple transgenic AD (3xTg-AD) mice, which progressively develop both amyloid and neurofibrillary tangle pathology, were vaccinated thrice with HSV(IE)Abeta(CMV)IL 4, or a set of control amplicon vectors. Increased Th2-related, Abeta-specific antibodies, improved learning and functioning of memory, and prevention of AD related amyloid and tau pathological progression were observed significantly more in the HSV(IE)Abeta(CMV)IL-4 vaccinated mice as compared to the other experimental groups. Our study underscores the potential of Abeta immunotherapy for AD and highlights the potency of amplicons in facilitating the immune response modulation to a disease-relevant antigen. PMID- 18388925 TI - A receptor-targeted nanocomplex vector system optimized for respiratory gene transfer. AB - Synthetic vectors for cystic fibrosis (CF) gene therapy are required that efficiently and safely transfect airway epithelial cells, rather than alveolar epithelial cells or macrophages, and that are nonimmunogenic, thus allowing for repeated delivery. We have compared several vector systems against these criteria including GL67, polyethylenimine (PEI) 22 and 25 kd and two new, synthetic vector formulations, comprising a cationic, receptor-targeting peptide K(16)GACSERSMNFCG (E), and the cationic liposomes (L) DHDTMA/DOPE or DOSEP3/DOPE. The lipid and peptide formulations self assemble into receptor-targeted nanocomplexes (RTNs) LED-1 and LED-2, respectively, on mixing with plasmid (D). LED-1 transfected airway epithelium efficiently, while LED-2 and GL67 preferentially transfected alveolar cells. PEI transfected airway epithelial cells with high efficiency, but was more toxic to the mice than the other formulations. On repeat dosing, LED-1 was equally as effective as the single dose, while GL67 was 30% less effective and PEI 22 kd displayed a 90% reduction of efficiency on repeated delivery. LED-1 thus was the only formulation that fulfilled the criteria for a CF gene therapy vector while GL67 and LED-2 may be appropriate for other respiratory diseases. Opportunities for PEI depend on a solution to its toxicity problems. LED-1 formulations were stable to nebulization, the most appropriate delivery method for CF. PMID- 18388926 TI - The host response to adenovirus, helper-dependent adenovirus, and adeno associated virus in mouse liver. AB - Understanding host responses to viral gene therapy vectors is necessary for the development of safe and efficacious in vivo gene transfer agents. We describe the use of high-density spotted complementary DNA microarrays in monitoring the in vivo host transcriptional responses in mouse liver upon administration of either a "first-generation"adenoviral (Ad) vector, a helper-dependent "gutless" adenoviral (HD) vector, or an adeno-associated viral (AAV) vector containing human factor IX (hFIX) expression cassettes. Since HD and AAV do not contain any viral genes, they allow us to assess the host response to the viral capsid and packaged nonviral DNA in whole animals. Comparison of the host response to Ad and HD helps assess the importance of leaky adenoviral gene expression. While all three vectors induced characteristic temporally sequenced programs of gene expression, the gene expression programs induced by the Ad and HD adenovirus vectors were remarkably similar, including the induction of a prominent type I interferon (IFN)-dependent cluster within 6 hours of administration. In contrast, the AAV-based vector caused far fewer alterations of host-gene expression. Our results indicate that recognition of the Ad capsid or double-stranded DNA (of nonviral origin) in the vector elicits a robust type I IFN response that is, however, not elicited by AAV-derived vector transduction. PMID- 18388927 TI - HIV TAT peptide modifies the distribution of DNA nanolipoparticles following convection-enhanced delivery. AB - We evaluated gene transfer using PEGylated bioresponsive nanolipid particles (NLPs) containing plasmid DNA administered by convection-enhanced delivery (CED) into orthotopically implanted U87-MG tumors in rat brain. We hypothesized that attachment of the human immunodeficiency virus trans-acting transcriptional activator peptide (TATp) to pH-sensitive, reduction-sensitive NLPs would increase gene transfer. TATp was attached either directly to a phospholipid (TATp-lipid) or via a 2-kd polyethylene glycol (PEG) to a lipid (TATp-PEG-lipid). Incorporation of 0.3 mol% TATp-PEG into pH-sensitive NLPs improved transfection 100,000-fold compared to NLPs in culture. In the brain or implanted tumors, the TATp-PEG restricted NLP convection to regions adjacent to the infusion catheter. Gene transfer in the brain from TATp-PEG NLPs, measured by green fluorescent protein (GFP) expression, was substantially greater than from NLPs adjacent to the catheter. Gene transfer using TATp-PEG NLPs, measured by luciferase expression, was 8-12-fold greater than from a 1,2-dioleoyl-3-trimethylammonium propane/cholesterol cationic lipoplex but 13-27-fold less than from the NLPs. Brain luciferase expression was localized in perivascular macrophages. Thus a cationic ligand, such as the TATp-PEG-lipid, can dramatically increase gene expression in culture, in the normal brain, and in implanted tumors; however, restriction of NLP distribution to the vicinity of the infusion catheter reduces the absolute level of gene transfer. PMID- 18388928 TI - Intron splicing-mediated expression of AAV Rep and Cap genes and production of AAV vectors in insect cells. AB - An artificial intron containing the insect cell polyhedrin (polh) promoter was designed, constructed, and inserted into the adeno-associated virus (AAV) Rep and Cap coding sequences to express the Rep and Cap proteins, respectively. The artificial intron was spliced out and full-length Rep78 or VP1 proteins were expressed from the insect promoters located upstream of their respective AUG start codons. The polh promoter located inside the artificial intron was functional, expressed the Rep52 or VP2/VP3 proteins located downstream of the artificial intron, and overlapped with the Rep78 or VP1 proteins. This is the first report that an artificial intron containing an insect cell promoter can be inserted into a coding sequence to express genes with overlapping open-reading frames (ORFs). A method was also established for AAV vector production in insect cells with these intron-containing Rep and Cap coding sequences, and the vectors produced thereby were infectious. These intron-containing AAV Rep and Cap coding sequences were very stable in recombinant baculoviruses and showed no apparent loss of protein expression even after five consecutive amplifications of the plaque-purified recombinant baculoviruses. This newly established AAV production method should prove to be a useful tool for large-scale AAV vector production. PMID- 18388929 TI - Therapeutic angiogenesis with intramuscular NV1FGF improves amputation-free survival in patients with critical limb ischemia. AB - This study evaluated the efficacy and safety of intramuscular administration of NV1FGF, a plasmid-based angiogenic gene delivery system for local expression of fibroblast growth factor 1 (FGF-1), versus placebo, in patients with critical limb ischemia (CLI). In a double-blind, randomized, placebo-controlled, European, multinational study, 125 patients in whom revascularization was not considered to be a suitable option, presenting with nonhealing ulcer(s), were randomized to receive eight intramuscular injections of placebo or 2.5 ml of NV1FGF at 0.2 mg/ml on days 1, 15, 30, and 45 (total 16 mg: 4 x 4 mg). The primary end point was occurrence of complete healing of at least one ulcer in the treated limb at week 25. Secondary end points included ankle brachial index (ABI), amputation, and death. There were 107 patients eligible for evaluation. Improvements in ulcer healing were similar for use of NV1FGF (19.6%) and placebo (14.3%; P = 0.514). However, the use of NV1FGF significantly reduced (by twofold) the risk of all amputations [hazard ratio (HR) 0.498; P = 0.015] and major amputations (HR 0.371; P = 0.015). Furthermore, there was a trend for reduced risk of death with the use of NV1FGF (HR 0.460; P = 0.105). The adverse event incidence was high, and similar between the groups. In patients with CLI, plasmid-based NV1FGF gene transfer was well tolerated, and resulted in a significantly reduced risk of major amputation when compared with placebo. PMID- 18388930 TI - Intralesional adenovirus-mediated interleukin-2 gene transfer for advanced solid cancers and melanoma. AB - Numerous preclinical and clinical studies have shown that interleukin-2 (IL-2) induces regression of metastatic tumors. We have conducted a phase I/II, multicenter, open-label, dose-escalating study to evaluate the safety, efficacy, and biological effects of repeated intratumoral injections of adenovirus-IL-2 (TG1024) in patients with advanced solid tumors and melanoma. Thirty five patients (twenty-five with metastatic melanoma and ten with other solid tumors) were treated in eight successive cohorts at dose levels ranging from 3 x 10(8) to 3 x 10(11) viral particles (vp). Intratumoral TG1024 injections in combination with dacarbazine (DTIC) were tested in metastatic melanoma in one cohort. No clinical responses were observed at doses below 3 x 10(11) vp. Six local objective responses were recorded in patients receiving 3 x 10(11) vp per treatment [five in metastatic melanoma and one in metastatic squamous cell carcinoma (SCC) of the skin], of which two were complete responses (CRs). Most of the common side effects were injection site reactions and flu-like syndrome. TG1024 dose intensification across cohorts resulted in increased serum IL-2 levels after the injection. Intratumoral TG1024 injection induced pronounced inflammation of the treated lesion, with predominant CD8(+), TIA+ lymphocytic infiltrate. Our results show that intratumoral injections of TG1024 are safe and well tolerated. The clinical activity of TG1024 observed in this study warrants further investigations. PMID- 18388931 TI - Double-blinded, Placebo-controlled plasmid GHRH trial for cancer-associated anemia in dogs. AB - The use of growth hormone releasing hormone (GHRH) plasmid-based therapy to treat companion dogs with spontaneous malignancies and anemia receiving a cancer specific treatment was examined in a double-blinded, placebo-controlled trial. The dogs (age 10.5 +/- 2.5 years, weight 24.9 +/- 12.9 kg) received a single 0.35 mg dose of plasmid or placebo intramuscularly (i.m.), followed by electroporation (EP), and were analyzed for up to 120 days. The response rate was defined as > or = 5% increase above the nadir in the red blood cell (RBC), hemoglobin (Hb), and hematocrit (Ht) levels. Plasmid-treated dogs had at least a 7% increase in all three parameters. The initial response rates for the plasmid-treated dogs were 40.6 and 35.5%, respectively on days 40 and 60, which increased to 54.2% on day 90. Although the response rate reduced to 47.1% by day 120, it was still 22.1% higher than in the control dogs. Post-hoc analysis of the GHRH-treated group showed that responder dogs survived 84% longer, 178 +/- 26 days post-treatment, while nonresponders and controls survived for 95 +/- 16 and 97 +/- 31 days post treatment, respectively. The quality of life, defined by 10 different parameters, dramatically improved with treatment. Overall, the possibility of a GHRH plasmid based therapy for anemia in cancer-afflicted subjects is important enough to deserve further investigation. PMID- 18388932 TI - Lentiviral vectors bearing the cardiac promoter of the Na+-Ca2+ exchanger report cardiogenic differentiation in stem cells. AB - Cardiosphere-derived resident cardiac stem cells (CDCs) are readily isolated from adult hearts and confer functional benefit in animal models of heart failure. To study cardiogenic differentiation in CDCs, we developed a method to genetically label and selectively enrich for cells that have acquired a cardiac phenotype. Lentiviral vectors achieved significantly higher transduction efficiencies in CDCs than any of the nine adeno-associated viral (AAV) serotypes tested. To define the most suitable vector system for reporting cardiogenic differentiation, we compared the cell specificity of five commonly-used cardiac-specific promoters in the context of lentiviral vectors. The promoter of the cardiac sodium-calcium exchanger (NCX1) conveyed the highest degree of cardiac specificity, as assessed by transducing seven cell types with each vector and measuring fluorescence intensity by flow cytometry. NCX1-GFP-positive CDC subpopulations, demonstrating prolonged expression of a variety of cardiac markers, could be isolated and expanded in vitro. Finally, we used chemical biology to validate that lentiviral vectors bearing the cardiac NCX1-promoter can serve as a highly accurate biosensor of cardiogenic small molecules in stem cells. The ability to accurately report cardiac fate and selectively enrich for cardiomyocytes and their precursors has important implications for drug discovery and the development of cell-based therapies. PMID- 18388933 TI - Intra-amniotic delivery of CFTR-expressing adenovirus does not reverse cystic fibrosis phenotype in inbred CFTR-knockout mice. AB - Due to its early onset and severe prognosis, cystic fibrosis (CF) has been suggested as a candidate disease for in utero gene therapy. In 1997, a study was published claiming that to how transient prenatal expression of CF transmembrane conductance regulator (CFTR) from an in utero-injected adenovirus vector could achieve permanent reversal of the CF intestinal pathology in adult CF knockout mice, despite the loss of CFTR transgene expression by birth. This would imply that the underlying cause of CF is a prenatal defect for which lifelong cure can be achieved by transient prenatal expression of CFTR. Despite criticism at the time of publication, no independent verification of this contentious finding has been published so far. This is vital for the development of future therapeutic strategies as it may determine whether CF gene therapy should be performed prenatally or postnatally. We therefore reinvestigated this finding with an identical adenoviral vector and a knockout CF mouse line (Cftr(tmlCam)) with a completely inbred genetic background to eliminate any effects due to genetic variation. After delivery of the CFTR-expressing adenovirus to the fetal mouse, both vector DNA and transgenic CFTR expression were detected in treated animals postpartum but statistically no significant difference in survival was observed between the Cftr(-/-) mice treated with the CFTR-adenovirus and those treated with the control vector. PMID- 18388934 TI - Adenovirus-mediated in utero expression of CFTR does not improve survival of CFTR knockout mice. AB - Gene therapy is being investigated in the treatment of lung-related aspects of the genetic disease, Cystic fibrosis (CF). Clinical studies have demonstrated CF transmembrane conductance regulator (CFTR) expression in the airways of adults with CF using a variety of gene transfer agents. In utero gene therapy is an alternative approach that facilitates vector transduction of rapidly expanding populations of target cells while avoiding immune recognition of the vector. In CF, in utero gene transfer could potentially delay the onset of disease symptoms in childhood and compensate for the role, if any, that CFTR plays in the developing organs. Previously published studies have suggested that transient expression of CFTR in utero was sufficient to rescue the fatal intestinal defect in S489X Cftr(tm1Unc)/Cftr(tm1Unc) knockout mice. We replicated these studies using an identical CFTR-expressing adenoviral vector and CF mouse strain in sufficiently large numbers to provide robust Kaplan-Meier survival data. Although each step of the procedure was carefully controlled and vector-specific CFTR expression was confirmed in the fetal organs after treatment, there was statistically no significant improvement in the survival of mice treated in utero with AdCFTR, compared with contemporaneous control animals. PMID- 18388935 TI - Remission of diabetes by beta-cell regeneration in diabetic mice treated with a recombinant adenovirus expressing betacellulin. AB - Type 1 diabetes results from destruction of the majority of the pancreatic beta cells by beta cell-specific autoimmune responses; therefore, expansion of the beta-cell mass in vivo is a possible approach to its treatment. Betacellulin (BTC) is known to promote beta-cell growth and differentiation. We investigated whether transient, constitutive expression, and secretion of BTC would regenerate sufficient numbers of pancreatic beta cells to restore normoglycemia in diabetic animals. We constructed a recombinant adenoviral vector (rAd-BTC) containing the cytomegalovirus promoter/enhancer, beta-globin chimeric intron, and albumin leader sequence to facilitate secretion, followed by BTC (1-80) complementary DNA (cDNA) encoding mature BTC. A single intravenous (i.v.) administration of rAd-BTC resulted in complete remission of streptozotocin (STZ)-induced diabetes within 2 weeks in mice. The mice remained normoglycemic for >100 days; glucose tolerance tests showed kinetics similar to normal, nondiabetic mice. Pancreatic insulin content, beta-cell mass, and serum insulin levels in rAd-BTC-treated mice were significantly higher than in the controls. Treatment of autoimmune diabetic mice with rAd-BTC in combination with an immune suppressor resulted in remission of diabetes. We conclude that transient expression of BTC by rAd-BTC administration results in prolonged remission of diabetes in mice, by the regeneration of sufficient numbers of beta cells in the pancreas. PMID- 18388937 TI - The prion organotypic slice culture assay--POSCA. AB - Methods enabling prion replication ex vivo are important for advancing prion science. However, few such technologies exist and many prion strains are intractable with them. Here, we describe a prion organotypic slice culture assay (POSCA), which allows for prion amplification and titration ex vivo under conditions that closely resemble intracerebral infection. Organotypic slices are incubated with infectious inoculum as free-floating sections, washed and cultured for up to 8 weeks. Slice cultures are a rich source of protein or RNA and allow for stringent comparisons between uninfected and prion-infected samples generated from the same mouse. Thirty-five days after contact with prions, cerebellar slices have amplified PrP(Sc) quantitatively similar to that seen in vivo, but accelerated fivefold. The POSCA detects replication of specific prion strains from disparate sources, including bovines and ovines, with variable efficiency. The culture procedure and prion infection can be performed in 8 h. PMID- 18388938 TI - MicroRNA expression profiling using microarrays. AB - Microarray technology is a powerful high-throughput tool capable of monitoring the expression of thousands of small noncoding RNAs at once within tens of samples processed in parallel in a single experiment. To conduct a genome-wide analysis of miRNA expression of normal and disease samples, such as cancer, and to distinguish expression signatures associated with diagnosis, prognosis and therapeutic interventions, we have developed a unique miRNA microarray assay on a CodeLink platform. The miRNA array consists of 4,104 probes printed in duplicate. This array can simultaneously profile more than 1,500 mature miRNAs and their corresponding precursors from 474 human and 373 mouse miRNA genes. The full protocol details of the miRNA microarray assay developed by our group are described here, including miRNA oligo probe design, array fabrication and miRNA target preparation (by reverse transcription of total RNA), target-probe hybridization on array, signal detection and data analysis. The assay is simple, can be easily standardized and allows the reproducible profiling of up to 24 total RNA samples within 24 h. PMID- 18388939 TI - Displacement of protein-bound aptamers with small molecules screened by fluorescence polarization. AB - Small molecule inhibitors of proteins are invaluable tools in research and as starting points for drug development. However, their screening can be tedious, as most screening methods have to be tailored to the corresponding drug target. Here, we describe a detailed protocol for a modular and generally applicable assay for the identification of small organic compounds that displace an aptamer complexed to its target protein. The method relies on fluorescence-labeled aptamers and the increase of fluorescence polarization upon their binding to the target protein. The assay has high Z'-factors, making it compatible with high throughput screening. It allows easy automation, making fluorescence readout the time-limiting step. As aptamers can be generated for virtually any protein target, the assay allows identification of small molecule inhibitors for targets or individual protein domains for which no functional screen is available. We provide the step-by-step protocol to screen for antagonists of the cytohesin class of small guanosine exchange factors. PMID- 18388940 TI - Visualization of protein interactions in living Caenorhabditis elegans using bimolecular fluorescence complementation analysis. AB - The bimolecular fluorescence complementation (BiFC) assay is a powerful tool for visualizing and identifying protein interactions in living cells. This assay is based on the principle of protein-fragment complementation, using two nonfluorescent fragments derived from fluorescent proteins. When two fragments are brought together in living cells by tethering each to one of a pair of interacting proteins, fluorescence is restored. Here, we provide a protocol for a Venus-based BiFC assay to visualize protein interactions in the living nematode, Caenorhabditis elegans. We discuss how to design appropriate C. elegans BiFC cloning vectors to enable visualization of protein interactions using either inducible heat shock promoters or native promoters; transform the constructs into worms by microinjection; and analyze and interpret the resulting data. When expression of BiFC fusion proteins is induced by heat shock, the fluorescent signals can be visualized as early as 30 min after induction and last for 24 h in transgenic animals. The entire procedure takes 2-3 weeks to complete. PMID- 18388941 TI - MRI of postmortem specimens of endangered species for comparative brain anatomy. AB - Many of the 5,500 threatened species of vertebrates found worldwide are highly protected and generally unavailable for scientific investigation. Here we describe a noninvasive protocol to visualize the structure and size of brain in postmortem specimens. We demonstrate its utility by examining four endangered species of kiwi (Apteryx spp.). Frozen specimens are thawed and imaged using MRI, revealing internal details of brain structure. External brain morphology and an estimate of brain volume can be reliably obtained by creating 3D models. This method has facilitated a comparison of brain structure in the different kiwi species, one of which is on the brink of extinction. This new approach has the potential to extend our knowledge of brain structure to species that have until now been outside the reach of anatomical investigation. PMID- 18388942 TI - Culture of Drosophila S2 cells and their use for RNAi-mediated loss-of-function studies and immunofluorescence microscopy. AB - Cultured Drosophila cell lines have become an increasingly popular model system for cell biological and functional genomic studies. One of the most commonly used lines, S2 cells, is particularly useful as it is easy to grow and maintain in the lab, is highly susceptible to gene inhibition using RNAi and is well suited to high-resolution light microscopic assays. Here, we provide protocols for the routine culture and RNAi treatment of S2 cells and methods to prepare these cells for fluorescence microscopy. Using these techniques, loss-of-function experiments may be performed after 4-7 d of RNAi-mediated protein depletion. PMID- 18388943 TI - Protocol for the induction of arthritis in C57BL/6 mice. AB - Collagen-induced arthritis is a well-validated, but strain-dependent mouse model of rheumatoid arthritis, with H-2(q) and H-2(r) strains showing the greatest degree of susceptibility. This protocol describes the induction of arthritis in the C57BL/6 strain (H-2(b)), which forms the genetic background of the majority of genetically modified strains. This protocol involves purification of type II collagen from chicken sternums, immunization of mice, clinical assessment of arthritis and analysis of T- and B-cell responses to type II collagen. Key aspects of the protocol are the need to use chicken collagen for immunization and the importance of avoiding aggressive behavior in males. The incidence of arthritis varies from 50 to 80% and is milder than the classical collagen-induced arthritis model. This procedure takes approximately 3 months to complete. PMID- 18388944 TI - Colocalization of fluorescent markers in confocal microscope images of plant cells. AB - This protocol describes the steps needed to perform quantitative statistical colocalization on two-color confocal images, specifically of plant cells. The procedure includes a calibration test to check the chromatic alignment of the confocal microscope. A software tool is provided to calculate the Pearson and Spearman correlation coefficients ('Pearson-Spearman correlation colocalization' ImageJ plug-in) across regions of interest within the image. Steps are included to help the user practice using the software. The result is a quantitative estimate of the amount of colocalization in the images. Manual masking takes about 1-15 min per image, depending on the detail required, and calculating the correlation coefficients is almost instantaneous. Examples of suitable dyes for such two-color colocalization include Oregon Green or Alexa Fluor 488 dyes in the green range (excited with 488-nm laser line) and Alexa Fluor 555 dye in the red range (excited with 543-nm laser line). PMID- 18388945 TI - Quantitative bioluminescence imaging of tumor-targeting bacteria in living animals. AB - We describe a protocol for imaging bacterial luciferase (Lux)-expressing bacteria in small living animals. In this protocol, light emitted by Lux-expressing bacteria is detected and monitored by a cooled charge-coupled device detector. When these bacteria are administered to animals, it provides a potentially valuable approach to generate sensitive whole-body images with extremely low background. This imaging technology should enable the real-time monitoring of bacterial migration into both primary and metastatic tumors in several different mouse tumor models at a strong quantification power. PMID- 18388946 TI - Detecting ligands and dissecting nuclear receptor-signaling pathways using recombinant strains of the yeast Saccharomyces cerevisiae. AB - This is a general protocol for the identification of natural and xenobiotic ligands of metazoan nuclear receptors (NRs) expressed in yeast. Yeast engineered to express an NR and a response element-driven reporter gene provide a system to detect and quantify ligand-dependent transcriptional activity. Such assays allow researchers to measure different types of ligands and determine dose-dependent activation of NRs. This methodology can also be used to examine the components of signal transduction pathways when conducted with mutant or engineered yeast strains expressing additional proteins or having alternate DNA response elements. This assay typically takes 2-3 d to complete, but most of this time entails cell growth rather than 'hands on' time. PMID- 18388947 TI - Solid-phase synthesis of pseudo-complementary peptide nucleic acids. AB - Pseudo-complementary peptide nucleic acid (pcPNA) is a DNA analog in which modified DNA bases 2,6-diaminopurine (D) and 2-thiouracil (U(s)) 'decorate' a poly[N-(2-aminoethyl)glycine] backbone, together with guanine (G) and cytosine (C). One of the most significant characteristics of pcPNA is its ability to effect double-duplex invasion of predetermined DNA sites inducing various changes in the biological and the physicochemical properties of the DNA. This protocol describes solid-phase synthesis of pcPNA. The monomers for G and C are commercially available, but the monomers for D and U(s) need to be synthesized (or can be ordered to custom synthesis companies). Otherwise, the procedure is the same as that employed for Boc-strategy synthesis of conventional PNA. This protocol, if the synthesis of D and U(s) monomers is not factored in, takes approximately 7 d to complete. PMID- 18388948 TI - Artificial restriction DNA cutter for site-selective scission of double-stranded DNA with tunable scission site and specificity. AB - The artificial restriction DNA cutter (ARCUT) method to cut double-stranded DNA at designated sites has been developed. The strategy at the base of this approach, which does not rely on restriction enzymes, is comprised of two stages: (i) two strands of pseudo-complementary peptide nucleic acid (pcPNA) anneal with DNA to form 'hot spots' for scission, and (ii) the Ce(IV)/EDTA complex acts as catalytic molecular scissors. The scission fragments, obtained by hydrolyzing target phosphodiester linkages, can be connected with foreign DNA using DNA ligase. The location of the scission site and the site-specificity are almost freely tunable, and there is no limitation to the size of DNA substrate. This protocol, which does not include the synthesis of pcPNA strands, takes approximately 10 d to complete. The synthesis and purification of the pcPNA, which are covered by a related protocol by the same authors, takes an additional 7 d, but pcPNA can also be ordered from custom synthesis companies if necessary. PMID- 18388949 TI - Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation. AB - Synchronization of cells and nuclei is a powerful technique for the exact study of regulatory mechanisms and for understanding cell cycle events. Counterflow centrifugal elutriation is a biophysical cell separation technique in which cell size and sedimentation density differences of living cells are exploited to isolate subpopulations in various stages of cell cycle. Here, a protocol is described for the separation of phase-enriched subpopulations from exponentially growing Chinese hamster ovary cells at high-resolution power of elutriation. The efficiency of elutriation is confirmed by measuring the DNA content fluorimetrically and by flow cytometry. The resolution power of elutriation is demonstrated by the ability to fractionate nuclei of murine pre-B cells. The installation and elutriation by collecting 16-30 synchronized fractions, including particle size analysis, can be achieved in 4-5 h. PMID- 18388950 TI - Three-dimensional lithographically defined organotypic tissue arrays for quantitative analysis of morphogenesis and neoplastic progression. AB - Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen gels. Epithelial cells are seeded into the cavities and covered with a second layer of collagen. The cells reorganize into hollow tissues corresponding to the geometry of the cavities. Patterned tissue arrays can be produced in 3-4 h and will undergo morphogenesis over the following 1-3 d. The protocol can easily be adapted to study a variety of tissues and aspects of normal and neoplastic development. PMID- 18388951 TI - NMR: prediction of molecular alignment from structure using the PALES software. AB - Orientational restraints such as residual dipolar couplings promise to overcome many of the problems that traditionally limited liquid-state nuclear magnetic resonance spectroscopy. Recently, we developed methods to predict a molecular alignment tensor and thus residual dipolar couplings for a given molecular structure. This provides many new opportunities for the study of the structure and dynamics of proteins, nucleic acids, oligosaccharides and small molecules. This protocol details the use of the software PALES (Prediction of AlignmEnt from Structure) for prediction of an alignment tensor from a known three-dimensional (3D) coordinate file of a solute. The method is applicable to alignment of molecules in many neutral and charged orienting media and takes into account the molecular shape and 3D charge distribution of the molecule. PMID- 18388952 TI - Facile synthesis of mono-6-amino-6-deoxy-alpha-, beta-, gamma-cyclodextrin hydrochlorides for molecular recognition, chiral separation and drug delivery. AB - We describe a protocol for the synthesis of mono-6-amino-6-deoxy-cyclodextrin hydrochloride (CD-NH3Cl), applicable to alpha-, beta- and gamma-cyclodextrin. These structurally simplest, highly water-soluble cationic cyclodextrins can be widely used in molecular recognition, chiral separation and drug delivery studies. Starting from commercially available chemicals, CD-NH3Cl is synthesized in four steps: (i) selective tosylation of cyclodextrin by the use of p toluenesulfonyl chloride to afford mono-6-(p-toluenesulfonyl)-6-deoxy cyclodextrin (Ts-CD); (ii) azide substitution of Ts-CD with sodium azide to afford mono-6-azido-6-deoxy-cyclodextrin (CD-N3); (iii) reduction of CD-N3 with triphenylphospine followed by hydrolysis to prepare mono-6-amino-6-deoxy cyclodextrin (CD-NH2); and (iv) treatment of CD-NH2 with hydrochloric acid to afford the titled CD-NH3Cl with good yield. The overall protocol requires approximately 2 weeks. PMID- 18388953 TI - Chromatin immunoprecipitation (ChIP) coupled to detection by quantitative real time PCR to study transcription factor binding to DNA in Caenorhabditis elegans. AB - In order to determine how signaling pathways differentially regulate gene expression, it is necessary to identify the interactions between transcription factors (TFs) and their cognate cis-regulatory DNA elements. Here, we have outlined a chromatin immunoprecipitation (ChIP) protocol for use in whole Caenorhabditis elegans extracts. We discuss optimization of the procedure, including growth and harvesting of the worms, formaldehyde fixation, TF immunoprecipitation and analysis of bound sequences through real-time PCR. It takes approximately 10-12 d to obtain the worm culture for ChIP; the ChIP procedure is spaced out over a period of 2.5 d with two overnight incubations. PMID- 18388954 TI - A protocol describing the principles of cis-regulatory analysis in the sea urchin. AB - cis-Regulatory analysis (CRA) is the precise identification of the cis-acting genomic sequences regulating gene transcription. As such, CRA provides essential mechanistic insight into key biological processes such as development. The first phase of this protocol involves identification of a large (approximately 100 kb) clone of genomic material surrounding the gene of interest and use of this clone to establish a reliable and unambiguous reporter assay. In the second phase, phylogenetic footprinting is used to identify candidate regulatory modules; these genomic sequences are then recursively tested for reporter activity. In the final phase, potential transcription factor binding sites are identified and disrupted in reporter constructs for individual testing. The strengths of this method reflect the use of large clones containing all relevant genomic regulatory sequences to establish a reporter assay with high fidelity. Given these foundational elements, predicted or suspected regulatory inputs can be rigorously tested and novel regulatory inputs identified. Although the expected time line varies greatly with the depth of information required, results may be obtained in as little as 4-6 months, but more detailed analyses will require several years to complete. PMID- 18388955 TI - Cardiac tissue engineering using perfusion bioreactor systems. AB - This protocol describes tissue engineering of synchronously contractile cardiac constructs by culturing cardiac cell populations on porous scaffolds (in some cases with an array of channels) and bioreactors with perfusion of culture medium (in some cases supplemented with an oxygen carrier). The overall approach is 'biomimetic' in nature as it tends to provide in vivo-like oxygen supply to cultured cells and thereby overcome inherent limitations of diffusional transport in conventional culture systems. In order to mimic the capillary network, cells are cultured on channeled elastomer scaffolds that are perfused with culture medium that can contain oxygen carriers. The overall protocol takes 2-4 weeks, including assembly of the perfusion systems, preparation of scaffolds, cell seeding and cultivation, and on-line and end-point assessment methods. This model is well suited for a wide range of cardiac tissue engineering applications, including the use of human stem cells, and high-fidelity models for biological research. PMID- 18388956 TI - A protocol for expression of foreign genes in chloroplasts. AB - Several major costs associated with the production of biopharmaceuticals or vaccines in fermentation-based systems could be minimized by using plant chloroplasts as bioreactors, which facilitates rapid scale-up. Oral delivery of chloroplast-derived therapeutic proteins through plant cells eliminates expensive purification steps, low temperature storage, transportation and sterile injections for their delivery. Chloroplast transformation technology (CTT) has also been successfully used to engineer valuable agronomic traits and for the production of industrial enzymes and biomaterials. Here, we provide a detailed protocol for the construction of chloroplast expression and integration vectors, selection and regeneration of transformants, evaluation of transgene integration and inheritance, confirmation of transgene expression and extraction, and quantitation and purification of foreign proteins. Integration of appropriate transgenes into chloroplast genomes and the resulting high levels of functional protein expression can be achieved in approximately 6 months in lettuce and tobacco. CTT is eco-friendly because transgenes are maternally inherited in most crop plants. PMID- 18388957 TI - NAD(P)H quinone oxidoreductase 1 inhibits the proteasomal degradation of the tumour suppressor p33(ING1b). AB - The tumour suppressor p33(ING1b) ((ING1b) for inhibitor of growth family, member 1b) is important in cellular stress responses, including cell-cycle arrest, apoptosis, chromatin remodelling and DNA repair; however, its degradation pathway is still unknown. Recently, we showed that genotoxic stress induces p33(ING1b) phosphorylation at Ser 126, and abolishment of Ser 126 phosphorylation markedly shortened its half-life. Therefore, we suggest that Ser 126 phosphorylation modulates the interaction of p33(ING1b) with its degradation machinery, stabilizing this protein. Combining the use of inhibitors of the main degradation pathways in the nucleus (proteasome and calpains), partial isolation of the proteasome complex, and in vitro interaction and degradation assays, we set out to determine the degradation mechanism of p33(ING1b). We found that p33(ING1b) is degraded in the 20S proteasome and that NAD(P)H quinone oxidoreductase 1 (NQO1), an oxidoreductase previously shown to modulate the degradation of p53 in the 20S proteasome, inhibits the degradation of p33(ING1b). Furthermore, ultraviolet irradiation induces p33(ING1b) phosphorylation at Ser 126, which, in turn, facilitates its interaction with NQO1. PMID- 18388958 TI - Intravitreal bevacizumab for macular edema secondary to retinal macroaneurysm. PMID- 18388959 TI - Should we anticipate intraoperative floppy iris syndrome (IFIS) even with very short history of tamsulosin? PMID- 18388960 TI - Tilted optic discs: The Beijing Eye Study. AB - PURPOSE: To determine the prevalence of tilted optic discs and their associations with ocular and general parameters. METHODS: The Beijing Eye Study included 4439 subjects out of 5324 subjects invited to participate (response rate 83.4%) with an age of 40+ years. The present investigation consisted of 4324 (97.4%) subjects for whom readable fundus photographs of at least one eye were available. The main outcome parameter was the presence of tilted optic discs defined as small discs with an oblique orientation and oval disc shape without signs of pathology in eyes without high myopia (defined as >-8 D). RESULTS: Tilted optic discs were detected in 31 eyes (0.36; 95% confidence interval: 0.23, 0.49%) of 23 patients (16 women). Tilted discs were associated with myopia (-6.59+/-0.68 D vs -0.60+/ 0.02 D, P<0.001), astigmatism (1.55+/-0.24 D vs 0.59+/-0.01 D, P<0.001), lower best corrected visual acuity (P<0.001), visual field defects (P<0.001), and small optic disc area (P<0.001). CONCLUSIONS: Tilted optic discs are present in about four out of 1000 eyes of adult Chinese in Northern China. They are associated with medium myopia, astigmatism, decreased visual acuity, and visual field defects. PMID- 18388961 TI - The evolving role of vascular endothelial growth factor inhibitors in the treatment of neovascular age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of blindness among the ageing population. The introduction of molecular inhibitors of vascular endothelial growth factor (VEGF), such as pegaptanib, ranibizumab, and bevacizumab, as treatments for exudative AMD has provided new hope for affected patients and has transformed the practices of retina specialists. Phase III clinical trials have demonstrated the efficacy and safety of monthly ranibizumab for the preservation as well as improvement of visual acuity in patients with exudative AMD. Ongoing trials are evaluating the effectiveness of different dosing regimens, monitoring strategies, and combination therapies to determine the optimal niche for this new class of drugs in AMD management. Based on emerging evidence, most clinicians are now adopting a variable VEGF inhibitor dosing strategy guided by serial diagnostic re-evaluation by optical coherence tomography. Some are also finding benefit through the addition of photodynamic therapy and steroids to the treatment regimen. The results of current and upcoming trials systematically addressing these issues are expected to establish new guidelines for the management of AMD. Indeed, a new paradigm may emerge wherein numerous modular therapeutic modalities are administered in customized combinations based on specific clinical and diagnostic findings. PMID- 18388963 TI - Surgery for bilateral macular holes. AB - OBJECTIVE: To report on bilateral sequential macular hole repair. DESIGN: Retrospective case series report. PARTICIPANTS: Sixteen eyes of eight patients with reduced visual acuity (VA) and metamorphopsia due to bilateral macular hole. INTERVENTION: Patients underwent vitrectomy surgery for bilateral macular hole. Seven patients had both eyes operated on consecutively at the same operating session. One patient had surgery to the fellow eye the next day. Strict facedown posturing was undertaken for 1 week by all patients. MAIN OUTCOME MEASURES: Closure of macular hole; final VA; complications. RESULTS: In 15 (94%) eyes the macular holes closed with resolution of symptoms; the mean VA at discharge was 6/15; two patients had reduced VA due to age-related macular degeneration (6/36, 6/18); five eyes had entry site tears; there was one case of lens touch with elevated intraocular pressure (IOP); in one (6%) patient the macular hole reopened after later cataract surgery (VA 6/24). CONCLUSIONS: Surgical closure of bilateral macular holes at the same operating session has distinct advantages but also considerable disadvantages. Case selection is paramount-patients require careful preoperative counselling and postoperative support, especially during the posturing week. PMID- 18388965 TI - Diagnostic nerve biopsy: a case history. PMID- 18388964 TI - Phase I trial of a pathotropic retroviral vector expressing a cytocidal cyclin G1 construct (Rexin-G) in patients with advanced pancreatic cancer. AB - Rexin-G is a pathotropic retroviral vector displaying a von Willebrand factor targeting motif and expressing a dominant negative cyclin G1 gene. We undertook a phase I trial of intravenous (i.v.) administration of Rexin-G in patients with gemcitabine refractory, metastatic pancreatic adenocarcinoma. Twelve patients were treated. Dose escalation was performed from a dose of 1 x 10(11) colony forming units (CFU) per cycle to 6 x 10(11) CFU per cycle. The treatment was well tolerated. One dose-limiting toxicity (DLT) at dose level 2 (1.5 x 10(11) CFU per cycle) was observed, consisting of grade 3 transaminitis. There was no detection of replication-competent virus in patients' peripheral blood mononuclear cells (PBMCs) or viral integration in DNA obtained from PBMCs, and no development of neutralizing antibodies. No evidence of antitumor activity was observed. The best objective response was progressive disease in 11 of the 12 study patients, while 1 patient showed radiographically stable disease with clinical deterioration and increase in the CA19.9 tumor marker. Median time to progression was 32 days. The median duration of survival of the study patients was 3.5 months from treatment initiation. Rexin-G is well tolerated in doses up to 6 x 10(11) CFU in patients with recurrent pancreatic cancer, but there was no evidence of clinical antitumor activity. PMID- 18388966 TI - Pathogenic potential of a collagenase gene from Aeromonas veronii. AB - The role of collagenase as a mechanism of bacterial pathogenicity in some pathogenic bacteria has been reported. The information on the role of collagenase in Aeromonas spp. pathogenesis is scant. In the present study, a mutant Aeromonas veronii RY001 that is deficient in the putative collagenase gene acg was constructed and compared with the wild-type strain for virulence factors. Bacterial cells and cell-free extracellular products of the mutant had significantly less collagenolytic activity, but there were not significant differences in caseinolytic, gelatinolytic, and elastolytic activities. Adhesion and invasion abilities of the mutant strain on epithelioma papillosum of carp cells was only 56% of that of the wild-type strain, and the cytotoxicity of the mutant strain to epithelioma papillosum of carp cells was only 42% of that of the wild-type strain. The LD50 values of the wild-type strain were determined as 1.6 x 10(6) and 3.5 x 10(5) cfu in goldfish and mice, respectively, whereas the mutant RY001 strain showed slightly higher values (i.e., 2.8 x 10(6) and 1.4 x 10(6) cfu in goldfish and mice, respectively). These results indicated the involvement of the collagenase gene in the pathogenesis of A. veronii. PMID- 18388967 TI - Effect of temperature on Brettanomyces bruxellensis: metabolic and kinetic aspects. AB - The effect of temperatures ranging from 15 to 35 degrees C on a culture of Brettanomyces bruxellensis was investigated in regards to thermodynamics, metabolism, and kinetics. In this temperature range, we observed an increase in growth and production rates. The growth behavior was well represented using the Arrhenius model, and an apparent activation energy of 16.61 kcal/mol was estimated. A stuck fermentation was observed at 35 degrees C as represented by high cell death. The carbon balance established that temperature had no effect on repartition of the glucose consumption between biomass and products. Hence, the same biomass concentration was obtained for all temperatures, except at 35 degrees C. Moreover, using logistic and Luedeking-Piret models, we demonstrated that production rates of ethanol and acetic acid were partially growth associated. Parameters associated with growth (alpha eth and alpha aa) remained constant with changing temperature, whereas, parameters associated with the population (beta eth and beta aa) varied. Optimal values were obtained at 32 degrees C for ethanol and at 25 degrees C for acetic acid. PMID- 18388968 TI - Comparative study of 7 fluorescent pseudomonad clinical isolates. AB - There is some debate about the potential survival of Pseudomonas fluorescens at temperatures above 37 degrees C and its consequences for infectious potential, owing to the heterogeneity of clinical strains. Seven clinical strains growing at 37 degrees C or more were submitted for polyphasic identification; 2 were identified as Pseudomonas mosselii and 4 were precisely characterized as P. fluorescens bv. I or II. The binding indexes on glial cells of the strains identified as P. fluorescens bv. I and P. mosselii were compared with that of a reference psychrotrophic strain, P. fluorescens MF37 (bv. V). Clinical P. fluorescens had a similar adherence potential range than strain MF37. Conversely, the binding indexes for P. mosselii strains were 3 times greater than that for strain MF37. These data, and those obtained by comparing the cytotoxic activities of P. fluorescens clinical strains, suggest the existence of different virulence mechanisms, leading either to a low infectious form or to a microorganism with cytotoxic activity in the same range as that of P. mosselii or even Pseudomonas aeruginosa. PMID- 18388969 TI - Differentiation of Sclerotinia minor depends on thiol redox state and oxidative stress. AB - Sclerotial differentiation in Sclerotinia minor is associated with oxidative stress and thiol redox state. The significance of oxidative stress to sclerotial differentiation was revealed by the higher oxidative stress of S. minor compared with a nonsclerotiogenic counterpart. The effect of thiol redox state on sclerotial differentiation was shown by the antioxidant action of the thiol (-SH) group of N-acetylcysteine and cysteine and by an unknown (not antioxidant) role of glutathione (GSH) on S. minor. The nonantioxidant role of GSH was indicated by the differentiation-inhibiting and differentiation-noninhibiting actions of the GSH biosynthesis inhibitor L-buthionine-S,R-sulfoximine and the GSH biosynthesis inducer L-2-oxo-thiazolidine-4-carboxylate, respectively, and by the increase of oxidative stress they caused during the transition from the undifferentiated to differentiated state of S. minor. Moreover, N-acetylcysteine can be used as a potent nontoxic fungicide against this phytopathogenic fungus by acting as a growth-inhibiting cytotoxic oxidant and by sustaining the fungus in the undifferentiated hyphal stage, which is vulnerable to degradation by soil microorganisms. PMID- 18388970 TI - Fibronectin enhances Campylobacter fetus interaction with extracellular matrix components and INT 407 cells. AB - Campylobacter fetus is a recognized pathogen of cattle and sheep that can also infect humans. No adhesins specific for C. fetus have to date been identified; however, bacterial attachment is essential to establish an infecting population. Scanning electron microscopy revealed C. fetus attachment to the serosal surface of human colonic biopsy explants, a location consistent with the presence of the extracellular matrix (ECM). To determine whether the ECM mediated C. fetus adherence, 7 C. fetus strains were assessed in a solid-phase binding assay for their ability to bind to immobilized ECM components. Of the ECM components assayed, adherence to fibronectin was noted for all strains. Attachment to ECM components was neither correlated with S-layer expression nor with cell-surface hydrophobicity. Ligand immunoblots, however, identified the S-layer protein as a major site of fibronectin binding, and modified ECM binding assays revealed that soluble fibronectin significantly enhanced the attachment of S-layer-expressing C. fetus strains to other ECM components. Soluble fibronectin also increased C. fetus adherence to INT 407 cells. This adherence was inhibited when INT 407 cells were incubated with synthetic peptides containing an RGD sequence, indicating that integrin receptors were involved in fibronectin-mediated attachment. Together, this data suggests that C. fetus can bind to immobilized fibronectin and use soluble fibronectin to enhance attachment to other ECM components and intestinal epithelial cells. In vivo, fibronectin would promote bacterial adherence, thereby, contributing to the initial interaction of C. fetus with mucosal and submucosal surfaces. PMID- 18388971 TI - Chromosomal gfp labelling of Pseudomonas aeruginosa using a mini-Tn7 transposon: application for studies of bacteria-host interactions. AB - Analysis of bacterial interactions with host cells using multiple techniques is essential for studies on microbial pathogenesis and for the development of new antimicrobial therapies. Pseudomonas aeruginosa is an important opportunistic pathogen that can cause severe, often life-threatening pulmonary infections in individuals with impaired host defense mechanisms. Using a mini-Tn7 transposon delivery system, we have chromosomally labelled the strain P. aeruginosa PAK with a green fluorescent protein gene (gfp) and tested PAKgfp as a research tool for studies of bacteria-host interactions. We were able to reliably and rapidly measure the interactions of PAKgfp with A549 human lung epithelial cells by using flow cytometry, a fluorometric microplate reader-based assay, and fluorescence microscopy. With these analytical tools, we have demonstrated the adhesion of PAKgfp to the extracellular matrix protein fibronectin and the involvement of fibronectin in PAKgfp-A549 cell interactions. PAKgfp can be successfully used to explore the effects of various pharmacological compounds on P. aeruginosa - host cell interactions in both in vitro and in vivo systems, with potentially important medical applications. PMID- 18388972 TI - Identification of Streptococcus pneumoniae genes specifically induced in mouse lung tissues. AB - To identify Streptococcus pneumoniae genes expressed specifically during infections, a selection system based on the in vivo expression technology (IVET) was established. galU, which is critical for capsular polysaccharide biosynthesis, and lacZY encoding beta-galactosidase were employed as dual reporter genes to screen in-vivo-induced (ivi) genes of S. pneumoniae. The galU deficient mutant of S. pneumoniae is incapable of utilizing galactose, thus failing to synthesize capsular polysaccharide, and therefore loses its ability to survive in the host. A promoter-trap library was constructed in S. pneumoniae, which was used to infect BALB/c mice in an intranostril model. Those strains recovered from lung tissue of mice and exhibiting a white colony phenotype on tryptic soy agar containing X-gal (5-bromo-4-chloro-3-indolyl-beta-D galactopyranoside) were collected and identificated. A total of 15 unique sequences were obtained through in vivo screening. The ivi genes of S. pneumoniae are involved in many processes, such as colonization and adherence, energy metabolism, nutrient substance transport, transcription regulation, DNA metabolism, and cell wall synthesis. There are some hypothetical proteins whose functions are not clear. This novel IVET is a useful tool for identifying ivi genes in S. pneumoniae. PMID- 18388973 TI - Repeated batch cultivation of the hydrocarbon-degrading, micro-algal strain Prototheca zopfii RND16 immobilized in polyurethane foam. AB - This study reports on the stability of the cells of a heterotrophic green micro algal strain Prototheca zopfii RND16 immobilized in polyurethane foam (PUF) cubes during degradation of mixed hydrocarbon substrate, which was composed of n alkanes and polycyclic aromatic hydrocarbons (PAHs), in 5 successive cycles of repeated batch cultivation at 30 degrees C. Both RND16 cells and mixed hydrocarbon substrate components had been entrapped in PUF cubes through cultivation. PUF-immobilized RND16 degraded n-alkanes almost completely, whereas the strain hardly degraded PAHs in PUFs, rather they accumulated in the matrices. It is noteworthy that this result is strikingly different from that of the free living cell culture, where RND16 reduced concentrations of both n-alkanes and PAHs. However, PAHs accumulation in the PUFs did not impair the performance of the immobilized alga to utilize n-alkanes. These results suggest that the PUFs harboring RND16 cells could be used repeatedly for selective retrieval of PAHs from oil-polluted waters after preferential biodegradation of n-alkanes by algae. PMID- 18388974 TI - Cloning of the ATP sulphurylase gene of Schizosaccharomyces pombe by functional complementation. AB - The ATP sulphurylase gene of Schizosaccharomyces pombe has been cloned by complementation of cysteine auxotrophy of a selenate-resistant mutant, which supposedly had a defect in ATP sulphurylase. A sulphate nonutilizing (cysteine auxotrophic) and selenate-resistant mutant of S. pombe was transformed with a wild-type S. pombe genomic library and sulphate-utilizing clones were isolated. The open reading frame encoding the ATP sulphurylase enzyme was found to be responsible for the restoration of sulphate assimilation. Transformants became as sensitive for selenate as the wild-type strain and produced a comparable amount of ATP sulphurylase as the prototrophic strains. The cloned ATP sulphurylase gene (sua1) proved to be an efficient selection marker in an ARS vector, when different isogenic or nonisogenic S. pombe selenate-resistant mutants were used as cloning hosts. Complementation of sua1- mutations by sua1-bearing multicopy vectors functions as a useful dual positive and negative selection marker. The cloned sua1 gene also complemented the met3 (ATP sulphurylase deficient) mutation in Saccharomyces cerevisiae. PMID- 18388975 TI - Properties of the reversible nonoxidative vanillate/4-hydroxybenzoate decarboxylase from Bacillus subtilis. AB - Bacillus subtilis (ATCC 6051) reversibly decarboxylates vanillate and 4 hydroxybenzoate under both aerobic and anoxic conditions. Thus, we have identified on the basis of gene sequence homology with Sedimentibacter hydroxybenzoicus and Streptomyces sp. strain D7, a putative B. subtilis hydroxybenzoate decarboxylase. The native form of this enzyme is encoded by 3 genes yclBCD (GI Sequence Identification Nos.: 2632649, 2632650, 2632651) that we have renamed during this research as bsdBCD to align with existing nomenclature. The bsdD gene is reported in the database to be 690 bp; however, our sequence analysis revealed that the size of this gene is in fact 228 bp, an observation that results in a shortening of YclD (i.e., BsdD) from 229 to 75 aa. The corresponding bsdBCD genes were cloned into Escherichia coli, and the heterologously expressed enzyme was assayed for activity. The decarboxylase exhibited a narrow substrate range, with only 2 of the tested substrates, vanillate (Kmapp = 4 mmol.L-1) and 4-hydroxybenzoate (Kmapp = ~1 mmol.L-1), being decarboxylated. The recombinant enzyme had properties similar to that of the native enzyme in respect to specific activity, kinetic properties, bidirectional decarboxylase-carboxylase activity, oxygen insensitivity, and substrate specificity. PMID- 18388976 TI - Comparison of treatment efficacy and stability of microbial populations between raw and anaerobically treated liquid pig manure, using PCR-DGGE and 16S sequencing. AB - The effects of adding an adapted inoculum to liquid pig manure (LPM) prior to anaerobic digestion were evaluated by standard analytical methods. In parallel, the phylogenetic diversity of the microbial community of raw and anaerobically digested pig manure was studied by both denaturing gradient gel electrophoresis (DGGE) and sequence analysis of 16S rRNA fragments amplified by polymerase chain reaction. Gas production, volative fatty acid production, removal of soluble chemical oxygen demand, and removal of volatile soluble solids were measured on raw and on inoculated liquid pig manure subjected to anaerobic digestion. DGGE profiles of 16S rRNA genes were used to compare the major elements of the bacterial community composition in raw LPM with those present under various incubation conditions. Major bands were excised and sequenced to gain insight into the identities of the bacterial populations from LPM treated under different conditions. The results show that the addition of an adapted inoculum did not have a major impact on the conversion of pig manure into soluble organic matter and did not significantly change the microbial populations present during anaerobic digestion of LPM. Bacterial composition also indicated that Clostridium species are important constituents of the LPM community. PMID- 18388977 TI - The influence of acetylsalicylic acid on oxylipin migration in Cryptococcus neoformans var. neoformans UOFS Y-1378. AB - In this paper we report the influence of acetylsalicylic acid on oxylipin migration in Cryptococcus neoformans var. neoformans UOFS Y-1378, previously isolated from human bone lesion. Transmission electron microscopy suggests that osmiophilic material originates in mitochondria and is deposited inside the yeast cell wall, from which it is excreted into the environment, along capsule protuberances, or through capsule detachments. Previous studies using immunogold labeling indicate that these osmiophilic layers contain 3-hydroxy oxylipins. In this study, the addition of acetylsalicylic acid (an inhibitor of mitochondrial function) in increasing amounts to the cells abrogated the migration of osmiophilic material, as well as capsule detachment from cell walls, and hence, oxylipin excretion. Consequently, we hypothesize that 3-hydroxy oxylipins are produced in mitochondria, probably via incomplete beta-oxidation or fatty acid synthesis, from which they are deposited inside the cell wall and excreted through tubular protuberances attached to the surrounding capsules and (or) through detachment of these oxylipin-containing capsules. PMID- 18388978 TI - Plesiomonas shigelloides hugZ encodes an iron-regulated heme binding protein required for heme iron utilization. AB - Plesiomonas shigelloides is an intestinal pathogen that uses heme as an iron source. The P. shigelloides heme utilization system consists of 10 genes, 7 of which permit heme transport and 3 of which are associated with utilization of heme as an iron source once it is inside the cell. The goal of this study was to examine hugZ, 1 of the 3 genes associated with utilization of heme iron. DPH8, a hugZ mutant, failed to grow to full cell density in media containing heme as the iron source, indicating that hugZ is required for heme iron utilization. Western blots using antibodies against Vibrio cholerae HutZ to detect the P. shigelloides HugZ indicated that hugZ encodes an iron-regulated cytoplasmic protein, which is absent in DPH8. A heme affinity bead assay performed on soluble protein fractions from P. shigelloides DPH8/pHUG24.5 (pHUG24.5 encodes hugZ) indicated that HugZ binds heme. Heme utilization was restored in DPH8 by hox1, which encodes the alpha-heme oxygenase from Synechocystis sp. strain PCC6803. However, HugZ did not exhibit alpha-heme oxygenase activity in an assay that detects the conversion of heme to the bilin functional group present in phycobiliproteins. These results do not rule out that HugZ exhibits another type of heme oxygenase activity not detected in the assay. PMID- 18388979 TI - Ultrastructural localization of heavy metals in the extraradical mycelium and spores of the arbuscular mycorrhizal fungus Glomus intraradices. AB - Arbuscular mycorrhizal fungi, obligate symbionts of most plant species, are able to accumulate heavy metals, thereby, protecting plants from metal toxicity. In this study, the ultrastructural localization of Zn, Cu, and Cd in the extraradical mycelium and spores of the arbuscular mycorrhizal fungus Glomus intraradices grown in monoxenic cultures was investigated. Zinc, Cu, or Cd was applied to the extraradical mycelium to final concentrations of 7.5, 5.0, or 0.45 mmol/L, respectively. Samples were collected at time 0, 8 h, and 7 days after metal application and were prepared for rapid freezing and freeze substitution. Metal content in different subcellular locations (wall, cytoplasm, and vacuoles), both in hyphae and spores, was determined by energy-dispersive X-ray spectroscopy. In all treatments and fungal structures analysed, heavy metals accumulated mainly in the fungal cell wall and in the vacuoles, while minor changes in metal concentrations were detected in the cytoplasm. Incorporation of Zn into the fungus occurred during the first 8 h after metal addition with no subsequent accumulation. On the other hand, Cu steadily accumulated in the spore vacuoles over time, whereas Cd steadily accumulated in the hyphal vacuoles. These results suggest that binding of metals to the cell walls and compartmentalization in vacuoles may be essential mechanisms for metal detoxification. PMID- 18388980 TI - Distribution of 3-hydroxy oxylipins and acetylsalicylic acid sensitivity in Cryptococcus species. AB - Using a well tested antibody specific for 3-hydroxy oxylipins, we mapped the presence of these oxylipins in selected Cryptococcus (Filobasidiella) species. Immunofluorescence microscopy studies revealed that these compounds are deposited on cell wall surfaces, appendages, and collarettes. In vitro studies revealed that growth of Cryptococcus species was inhibited by acetylsalicylic acid (which is known to inhibit mitochondrial function, including the production of 3-hydroxy oxylipins) at concentrations as low as 1 mmol/L. The results suggest that acetylsalicylic acid is effective in controlling the growth of tested pathogens, probably by targeting their mitochondria. This study further expands the known function of this anti-inflammatory drug as anti-fungal agent. PMID- 18388981 TI - Identification and characterization of an alternative oxidase in the entomopathogenic fungus Metarhizium anisopliae. AB - Mitochondria of Metarhizium anisopliae contain an alternative oxidase (AOX), which reduces oxygen to water by accepting electrons directly from ubiquinol. AOX activity is demonstrated in situ as a constitutive enzyme. Greatest activity of AOX appears at the beginning and at the end of the fungal developmental cycle, germination of aerial conidia and the formation of submerged conidia, respectively. Changes in nutritional conditions, e.g., the presence of host insect cuticle or nutrient starvation had no effect on the induction of AOX activity. Antimycin A, an electron transport chain inhibitor, induced AOX activity. Cloning of the AOX DNA and the alignment of the deduced amino acid sequence of a segment of the AOX gene from M. anisopliae shows structural similarities with other AOX sequences with differing levels of variation when compared with homologous sequences from plants, yeasts, and filamentous fungi. Alternative oxidase in entomopathogenic fungi may have a positive contribution to ecological fitness. PMID- 18388982 TI - Transcriptional regulation of ACC deaminase gene expression in Pseudomonas putida UW4. AB - One of the major mechanisms that plant growth-promoting bacteria use to facilitate plant growth is through the lowering of plant ethylene levels by the bacterial enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Many of the bacterial ACC deaminase genes (acdS) that have been examined to date are under the transcriptional control of a leucine-responsive regulatory protein, Lrp, encoded by acdR and referred to here as AcdR. The work presented here is focused on how AcdR and the newly discovered AcdB protein from Pseudomonas putida UW4 are involved in the regulation of acdS expression. First, the results of gel retardation experiments showed that AcdR binds to the acdS regulatory region, and this binding activity in vitro is not affected by the addition of 2 mmol x L-1 ACC but can be eliminated by addition of 20 microg x mL-1 leucine. Second, a potential regulatory protein, AcdB, involved in the regulation of acdS expression, was identified through both yeast 2-hybrid screen and coimmunoprecipitation based on its ability to bind to AcdR; subsequently, its binding to the acdS regulatory region in the presence of ACC was shown by gel retardation experiments. The data are interpreted in terms of a model in which AcdR and AcdB co-regulate the expression of the acdS gene. PMID- 18388983 TI - Gene expression of the arsenic resistance operon in Chromobacterium violaceum ATCC 12472. AB - Chromobacterium violaceum ATCC 12472 presents an arsRCB-type operon, which is involved in arsenic resistance. The regulating protein of this resistance system (ArsR) does not have the small conserved site (ELCVDCL) to link to the metalloid, as observed in Escherichia coli, and is thus considered to be an atypical ArsR protein, like that observed in Acidithiobacillus ferrooxidans. In the present study, the gene expression profile of the ars operon under induction at different concentrations of arsenite - As(III) - was obtained via real-time PCR (TaqMan), by correlating the threshold cycle (Ct) values of induced and uninduced (control) samples. Through linear regression analysis (R2 = 0.9926), the gene expression profile of the ars operon showed clearly that the 0.125 micromol/L concentration of As(III) was sufficient to provoke a 4-fold increase in the resistance system, and a further increase in concentration resulted in an increase of up to 53-fold in transcription rates. The relation between resistance and induction of the ars operon indicates that the increased resistance to As(III) is associated with the increase in the number of transcripts. PMID- 18388984 TI - Large chromosomal inversion correlated with spectinomycin resistance in Lactococcus lactis subsp. lactis bv. diacetylactis S50. AB - A large chromosomal inversion that confers resistance to high concentrations of the antibiotic spectinomycin in Lactococcus lactis subsp. lactis bv. diacetylactis S50 was identified by pulsed field gel electrophoresis. The same type of inversion was identified in 4 independent experiments and in 4 different derivatives of strain S50, indicating the same position and the same mechanism of recombination as a response to antibiotic selective pressure in all derivatives. An analysis of ribosomal operons in strain S50 and mutants revealed that ribosomal operons are not endpoints of the recombination. Spectinomycin-resistant mutants appeared in a population of S50 derivatives at a high frequency of 2 x 10(-7). These spectinomycin-resistant mutants were not able to compete successfully with the wild-type strain during 25 generations (48 h) of co-culture in vitro, indicating that inversion had a significant fitness cost. Results demonstrate that as a mechanism of genome plasticity, inversion can be directly involved in one-step development of the adaptation to a high concentration of spectinomycin. PMID- 18388985 TI - Apoptosis induced by lipid-associated membrane proteins from Mycoplasma penetrans is mediated by nuclear factor kappaB activation in mouse macrophage. AB - Mycoplasma penetrans was shown to be involved in alteration of several eukaryotical cells functions and a causative agent in urogenital infectious diseases. Lipid-associated membrane proteins (LAMPs) may be responsible for the pathogenicity of some mycoplamas. In this study, we investigated whether M. penetrans LAMPs have pathogenic potential by inducing apoptosis in mouse macrophages. As analyzed by annexin-V - fluorescein isothiocyanate staining, significant early- and late-stage apoptosis was induced in M. penetrans LAMPs challenged mouse macrophages. And agarose gel electrophoresis of the DNA of M. penetrans LAMPs-challenged cells revealed a ladder-like pattern of migration of DNA indicative of apoptosis. The possible molecular mechanisms responsible for the induction of apoptosis were also investigated by characterizing the activation of nuclear transcription factor kappaB (NFkappaB). NFkappaB was activated and translocated into the nucleus in mouse macrophages stimulated by M. penetrans LAMPs. The activation of NFkappaB and M. penetrans LAMPs-induced apoptosis in mouse macrophages was partially inhibited by the NFkappaB-specific inhibitor pyrrolidine dithiocarbamate. Thus, this study demonstrates that M. penetrans LAMPs may be an important etiological factor owing to their ability to induce apoptosis in mouse macrophages, which is probably mediated through the activation of NFkappaB. PMID- 18388986 TI - A simple restriction fragment PCR approach for discrimination of humanpathogenic Old World animal Orthopoxvirus species. AB - There are reliable polymerase chain reaction assays available for exclusion of Variola virus from other poxviruses. However, the discrimination of humanpathogenic animal Orthopoxviridae is more challenging because of the high genomic conservation. Based on the variability of the A36R gene, we describe a simple 20 min PCR assay followed by a 1 h digest with 3 different restriction enzymes. This assay enables rapid discrimination between Cowpox virus and Monkeypox virus and discrimination of the most prevalent members of the Vaccinia virus and Camelpox virus. The test was orthopoxvirus specificand did not react with parapox (Orf) virus. Moreover, the amplified fragments were also well suited for additional genotyping by direct DNA sequencing. PMID- 18388987 TI - Hydroxamate siderophores produced by Streptomyces acidiscabies E13 bind nickel and promote growth in cowpea (Vigna unguiculata L.) under nickel stress. AB - The siderophore-producing ability of nickel-resistant Streptomyces acidiscabies E13 and the role of the elicited siderophores in promoting plant growth under iron and nickel stress are described. Siderophore assays indicated that S. acidiscabies E13 can produce siderophores. Electrospray ionization mass spectrometry (ESI-MS) revealed that the bacterium simultaneously produces 3 different hydroxamate siderophores. ESI-MS showed that in addition to iron, all 3 siderophores can bind nickel. In vitro plant growth tests were conducted with cowpea (Vigna unguiculata) in the presence and absence of the elicited siderophores. Culture filtrates containing hydroxamate siderophores significantly increased cowpea height and biomass, irrespective of the iron status of the plants, under nickel stress. The presence of reduced iron was found to be high in siderophore-containing treatments in the presence of nickel. Measurements of iron and nickel contents of cowpea roots and shoots indicated that the siderophore mediated plant growth promotion reported here involves the simultaneous inhibition of nickel uptake and solubilization and supply of iron to plants. We conclude that hydroxamate siderophores contained in culture filtrates of S. acidiscabies E13 promoted cowpea growth under nickel contamination by binding iron and nickel, thus playing a dual role of sourcing iron for plant use and protecting against nickel toxicity. PMID- 18388988 TI - Diversity of predominant endophytic bacteria in European deciduous and coniferous trees. AB - The diversity of endophytic bacteria residing in root, stem, and leaf tissues was examined in coniferous and deciduous tree species, Scots pine (Pinus sylvestris L.), silver birch (Betula pendula Roth), and rowan (Sorbus aucuparia L.). Using cultivation-dependent and -independent analyses, the bacterial communities were observed to be significantly different in the belowground (roots and rhizosphere) and aboveground (leaves and stems) samples of the respective host trees. No significant differences, with respect to the different tree species, were observed in the associated communities. Predominant cultivable endophytes isolated included bacteria closely related to Bacillus subtilis, Bacillus licheniformis, Paenibacillus spp., and Acinetobacter calcoaceticus. Comparisons of the most abundant cultivable bacteria in the rhizosphere and root samples suggested that root endophytic bacteria may be in residence through processes of selection or active colonization rather than by passive diffusion from the rhizosphere. PMID- 18388989 TI - Properties of a non collagen-degrading Bacillus subtilis keratinase. AB - Bacillus subtilis S14 produces a keratinase (KerS14) with non collagen-degrading activity. Indeed, this is the first keratinase described so far that does not have any detectable effect on collagen, which is a crucial property for an enzyme intended to be used in skin dehairing. Because of its importance as an industrial tanning enzyme, we report the biochemical characterization of KerS14. This protein exhibited an apparent molecular mass of 27 kDa, a pI of 6.5, and an optimum pH in the range of 8.0-9.0. The enzyme's activity was stimulated by Mn2+ (7.7-fold), Ca2+ (6.1-fold), Mg2+ (4.9-fold), and Co2+ (4.0-fold) but was inhibited by Cu2+ and Zn2+. Using p-nitroanilide and methylcoumarine derivatized peptides, we observed that KerS14 prefered Arg at subsite P1, small amino acid residues at subsite P2, and Gln or Glu at subsite P3. KerS14 presented higher keratin degradation specificity than other commercial proteases. Its high keratinolytic activity and the absence of virtually any activity against collagen remark the biotechnological potential of this enzyme to be used at larger scales in tannery dehairing processes. PMID- 18388990 TI - Microarray analysis of Streptococcus pneumoniae gene expression changes to human lung epithelial cells. AB - Streptococcus pneumoniae infection starts from the respiratory tract where interaction with host epithelial cells occurs. To gain more insights on pneumococcal pathogenesis, an oligonucleotide (oligo)-based microarray was used to investigate gene expression changes of one serotype 3 encapsulated pathogenic S. pneumoniae strain 82 and one unencapsulated avirulent S. pneumoniae strain R6 upon exposure to human lung epithelial cells (A549) for 1 and 3 h, respectively. We observed that genes associated with many functional categories were differentially regulated in strain 82, such as genes in pathogenesis, cell envelope, transcription, translation, transport, metabolism, and unknown functions. In contrast, few genes were changed in strain R6 except for genes in ribonucleotide biosynthesis and unknown functions. Quantitative real-time PCR analysis confirmed the microarray results for most of the genes tested. To further characterize functions of the selected genes, knockout mutants were constructed in strain R6. We demonstrated that 2 genetic loci, SP_2170 (AdcB, zinc ABC transporter) and SP_0157 (hypothetical protein), were involved in adherence to A549 cells. These data suggest that divergent gene expression changes occur in S. pneumoniae pathogenic and avirulent strains during interaction with human lung epithelial cells. Some of those genes are involved in pneumococcal pathogenesis. PMID- 18388992 TI - Phylogenetic and symbiotic characterization of rhizobial bacteria nodulating Argyrolobium uniflorum in Tunisian arid soils. AB - Forty-two bacterial isolates from root nodules of Argyrolobium uniflorum growing in the arid areas of Tunisia were characterized by phenotypic features, RFLP, and sequencing of PCR-amplified 16S rRNA genes. The isolates were found to be phenotypically diverse. The majority of the isolates tolerated 3% NaCl and grew at temperatures up to 40 degrees C. Phylogenetically, the new isolates were grouped in the genera Sinorhizobium (27), Rhizobium (13), and Agrobacterium (2). Except for the 2 Agrobacterium isolates, all strains induced nodulation on Argyrolobium uniflorum, but the number of nodules and nitrogen fixation efficiency varied among them. Sinorhizobium sp. strains STM 4034, STM 4036, and STM 4039, forming the most effective symbiosis, are potential candidates for inoculants in revegetalisation programs. PMID- 18388991 TI - Recovery of useful traits from isolates inhabiting an agricultural soil cultivated with herbicide-resistant poplars. AB - The aim of this study was to investigate the culturable bacteria living in soil cultivated with Basta-tolerant transgenic white poplars (Populus alba L. 'Villafranca'). Plate Count Agar medium containing phosphinothricin, the active component of Basta, was used to isolate the herbicide-resistant bacteria (HRB). No significant changes in the size of the soil microbial flora following herbicide treatment were observed. The characterization of HRB isolates by 16S rDNA-based taxonomy revealed a predominance of Pseudomonas and Bacillus species. The screening carried out on soil samples allowed for the recovery of isolates with useful properties for biotechnological and agronomical purposes, particularly in relation to root development. Among the tested isolates, only HRB 1b, HRB-1c, and HRB-7 showed remarkable swarming ability, a valuable trait supporting the beneficial plant-microbe interactions. HRB-1c was also characterized by consistent production of indoleacetic acid (17.8 +/- 0.09 microg x mL-1 x (OD600 unit)-1), and it was able to stimulate the in vitro growth of Villafranca explants. Since novel tools are constantly required to enhance productivity of perennial species and to expand their use for practical purposes, the availability of bacteria that support tree growth, such as the HRB-1c isolate, represents a significant advantage. PMID- 18388993 TI - Diversity and dynamics of lactobacilli populations during ripening of RDO Camembert cheese. AB - The diversity and dynamics of Lactobacillus populations in traditional raw milk Camembert cheese were monitored throughout the manufacturing process in 3 dairies. Culture-dependent analysis was carried out on isolates grown on acidified de Man - Rogosa - Sharpe agar and Lactobacillus anaerobic de Man Rogosa Sharpe agar supplemented with vancomycin and bromocresol green media. The isolates were identified by polymerase chain reaction - temperature gradient gel electrophoresis (PCR-TGGE) and (or) species-specific PCR and (or) sequencing, and Lactobacillus paracasei and Lactobacillus plantarum isolates were characterized by pulsed field gel electrophoresis (PFGE). Milk and cheese were subjected to culture-independent analysis by PCR-TGGE. Presumed lactobacilli were detected by plate counts throughout the ripening process. However, molecular analysis of total DNA and DNA of isolates failed to detect Lactobacillus spp. in certain cases. The dominant species in the 3 dairies was L. paracasei. PFGE analysis revealed 21 different profiles among 39 L. paracasei isolates. Lactobacillus plantarum was the second most isolated species, but it occurred nearly exclusively in one dairy. The other species isolated were Lactobacillus parabuchneri, Lactobacillus fermentum, Lactobacillus acidophilus, Lactobacillus helveticus, a Lactobacillus psittaci/delbrueckii subsp. bulgaricus/gallinarum/crispatus group, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, Lactobacillus brevis, Lactobacillus kefiri, and Lactobacillus perolens. Lactobacilli diversity at the strain level was high. Dynamics varied among dairies, and each cheese exhibited a specific picture of species and strains. PMID- 18388994 TI - Immunochemical studies and genetic background of two Neisseria meningitidis isolates expressing unusual capsule polysaccharide antigens with specificities of both serogroup Y and W135. AB - We described 2 unusual Neisseria meningitidis strains isolated from epidemiologically unrelated invasive meningococcal disease cases in Ontario, Canada. Both isolates have features typical of serogroup Y N. meningitidis: are of serotype 2c, are of the multi-locus sequence types typical of the serogroup Y strains in Canada, and are genotyped as serogroup Y based on a previously described PCR-ELISA method that detects the serogroup-Y-specific siaD gene. However, both strains were poly-agglutinable in both anti-Y and anti-W135 antisera. Further studies on 1 of these 2 isolates showed the presence of glucose and galactose as well as sialic acids in its purified capsular polysaccharide, suggesting the presence of both serogroup Y and serogroup W135 polysaccharides. Rabbit antisera produced to this strain contained antibodies to both purified serogroup Y and serogroup W135 capsular polysaccharides. Absorption experiments with either serogroup Y or serogroup W135 bacteria confirmed the presence of antibodies to these 2 different polysaccharides. DNA sequencing of the cps operon from both isolates revealed a siaD gene with 99.7% homology to the published siaD sequence from a serogroup Y strain but with 3 point mutations that all resulted in amino acid changes. How these strains may affect results of routine surveillance, PCR diagnosis, and immuno-protection by vaccination are discussed. PMID- 18388995 TI - Effect of an ntrC mutation on amino acid or urea utilization and on nitrogenase switch-off in Herbaspirillum seropedicae. AB - Herbaspirillum seropedicae is a nitrogen-fixing bacterium that grows well with ammonium chloride or sodium nitrate as alternative single nitrogen sources but that grows more slowly with L-alanine, L-serine, L-proline, or urea. The ntrC mutant strain DCP286A was able to utilize only ammonium or urea of these nitrogen sources. The addition of 1 mmol.L-1 ammonium chloride to the nitrogen-fixing wild type strain inhibited nitrogenase activity rapidly and completely. Urea was a less effective inhibitor; approximately 20% of nitrogenase activity remained 40 min after the addition of 1 mmol x L-1 urea. The effect of the ntrC mutation on nitrogenase inhibition (switch-off) was studied in strain DCP286A containing the constitutively expressed gene nifA of H. seropedicae. In this strain, nitrogenase inhibition by ammonium was completely abolished, but the addition of urea produced a reduction in nitrogenase activity similar to that of the wild-type strain. The results suggest that the NtrC protein is required for assimilation of nitrate and the tested amino acids by H. seropedicae. Furthermore, NtrC is also necessary for ammonium-induced switch-off of nitrogenase but is not involved in the mechanism of nitrogenase switch-off by urea. PMID- 18388996 TI - attG and attC mutations of Agrobacterium tumefaciens are dominant negative mutations that block attachment and virulence. AB - The cryptic plasmid (pAT) of Agrobacterium tumefaciens was not required for virulence or attachment to plant surfaces. However, mutations in the attC and attG genes located on pAT caused the bacteria to become avirulent and non attaching on tomato, carrot, and Bryophyllum daigremontiana. This was the case whether the mutation was in the copy of the genes located on pAT or whether it was carried in a second copy of the attA-G operon located on a plasmid in cells that contained a wild-type copy of pAT. Thus attC and attG mutations are dominant negative mutations. The mechanism by which these mutations block attachment and virulence is unknown. PMID- 18388997 TI - Isolation, selection, and characterization of beneficial rhizobacteria from pea, lentil, and chickpea grown in western Canada. AB - The use of beneficial soil microorganisms as agricultural inputs for improved crop production requires selection of rhizosphere-competent microorganisms with plant growth-promoting attributes. A collection of 563 bacteria originating from the roots of pea, lentil, and chickpea grown in Saskatchewan was screened for several plant growth-promoting traits, for suppression of legume fungal pathogens, and for plant growth promotion. Siderophore production was detected in 427 isolates (76%), amino-cyclopropane-1-carboxylic acid (ACC) deaminase activity in 29 isolates (5%), and indole production in 38 isolates (7%). Twenty-six isolates (5%) suppressed the growth of Pythium sp. strain p88-p3, 40 isolates (7%) suppressed the growth of Fusarium avenaceum, and 53 isolates (9%) suppressed the growth of Rhizoctonia solani CKP7. Seventeen isolates (3%) promoted canola root elongation in a growth pouch assay, and of these, 4 isolates promoted the growth of lentil and one isolate promoted the growth of pea. Fatty acid profile analysis and 16S rRNA sequencing of smaller subsets of the isolates that were positive for the plant growth-promotion traits tested showed that 39%-42% were members of the Pseudomonadaceae and 36%-42% of the Enterobacteriaceae families. Several of these isolates may have potential for development as biofertilizers or biopesticides for western Canadian legume crops. PMID- 18388998 TI - Production of cercosporin toxin by the phytopathogenic Cercospora fungi is affected by diverse environmental signals. AB - Cercosporin is a polyketide phytotoxin produced by many phytopathogenic Cercospora spp. We investigated environmental signals that have elaborate control of cercosporin production. Light is the most critical factor for cercosporin production. Cercospora nicotianae accumulated substantial quantities of cercosporin only when grown on a particular potato dextrose agar under light but produced little cercosporin on other brands of potato dextrose agar or media with defined ingredients. In addition to light regulation, numerous factors including salts, buffers, and ions markedly affected cercosporin production. By contrast, pH had little effect on cercosporin production. Depletion or alteration of the carbon or nitrogen sources also affected cercosporin production. Production of cercosporin was elevated to varying levels by metal ions, such as cobalt, ferric, manganese, and zinc. Significant differences in cercosporin production were observed among various Cercospora species. Further, regulation of cercosporin production by phosphate buffer, ammonium, LiCl, but not metal ions appeared to occur at transcriptional levels. Expression of the genes involved in cercosporin biosynthesis and regulation decreased markedly and was closely concomitant with the amounts of cercosporin reduced as the fungus was grown on medium containing phosphate, LiCl, ammonium, or dimethyl sulfoxide. The results reveal the complexity of cercosporin production at the physiological and genetic levels. A model delineating regulatory controls of cercosporin biosynthesis is proposed and discussed. PMID- 18388999 TI - Diversity and distribution of the prokaryotic community in near-surface permafrost sediments in the Tianshan Mountains, China. AB - The community structures and diversity of bacteria and archaea were investigated at 4 depths (1.5, 2.0, 2.5, and 3.0 m) in permafrost sediments in the Tianshan Mountains, using denaturing gradient gel electrophoresis of 16S rRNA gene amplified by polymerase chain reaction. Phylogenetic analysis of the dominant bands sequenced revealed the presence of rich diversity of bacteria, which could be related to the Proteobacteria, Actinobacteria, Acidobacteria, Gemmatimonadetes, Bacteroidetes, Firmicutes, and Chloroflexi. The Proteobacteria, consisting of the alpha, beta, gamma and epsilon subdivisions, were clearly the dominant group at all depths studied. Archaeal diversity was relatively low and archaeal 16S rRNA gene sequences were grouped into 3 phylogenetic clusters within the 2 kingdoms Euryarchaeota and Crenarchaeota. Within the Euryarchaeota, methanogen-related group II was most abundant at shallow depth (1.5 m), whereas halobacterium-related group I dominated at greater depths. The low-temperature Crenarchaeota group was detected only at 2.5 and 3.0 m. Specific-depth distribution of methanogen-related Euryarchaeota group II and denitrifying bacteria of the genus Pseudomonas dominated at 1.5 m depth, accompanied by a distinct peak in the ratio of NH4-N to NO3/NO2-N, implying the potential capacity of these organisms in near-surface permafrost to release the greenhouse gases N2O and CH4. PMID- 18389000 TI - Structural elucidation of lipopolysaccharide core oligosaccharides from lic1 and lic1/lic2 mutants of Haemophilus influenzae type b strain Eagan. AB - The structures of lipopolysaccharides (LPSs) of lic1 and lic1/lic2 mutants from Haemophilus influenzae type b strain Eagan (RM153) were investigated using methylation analysis, electrospray ionization - mass spectrometry, and nuclear magnetic resonance spectroscopy on O-deacylated, O- and N-deacylated core oligosaccharide (OS); and deacylated, dephosphorylated, and terminally reduced samples. The backbone OS derived from the major LPS glycoforms were determined to consist of the inner-core triheptosyl unit, L-alpha-D-Hepp-(1-2)-L-alpha-D-Hepp (1-3)-L-alpha-D-Hepp-(1-, common to all H. influenzae strains investigated to date that is linked to the lipid A region of the molecule via a Kdo residue to which beta-D-Glcp and beta-D-Galp residues are attached in 1,4 and 1,2 linkages to the proximal (HepI) and distal (HepIII) heptose residues, respectively. It was found that the lic1 mutant predominately elaborates the Hex4 LPS glycoforms previously identified in the parent strain where a beta-D-Glcp-(1-4)-alpha-D-Glcp unit is linked in a 1,3 linkage to the central heptose (HepII) of the triheptosyl moiety. The lic1 locus consists of 4 genes (lic1A to lic1D) in a single transcriptional unit that directs phase variable expression of phosphocholine. The lic1A gene is phased off in the RM153 isolate of strain Eagan. LPS from the double mutant, lic1/lic2 had a similar structure to that of lic1 mutant except that there was no chain extension from the central heptose in the inner core (HepII). The lic2 locus consists of 4 genes (lic2A to lic2D). Our structural data were consistent with the proposed function of lic2C, providing the first definitive evidence for its role as the glycosyltransferase required for chain initiation from HepII. The presence of an O-acetyl group at O-3 of the distal heptose (HepIII) was elucidated by 1H NMR on the mild acid liberated core OS samples. PMID- 18389001 TI - Ca2+-dependent induction of conidiation in submerged cultures of Trichoderma viride. AB - The presence of Ca2+ (up to 0.1 mol/L) in the cultivation media was found to induce the formation of conidia in submerged mycelia of Trichoderma viride in a concentration-dependent manner. Ca2+ dramatically stimulated conidiation after 70 h of cultivation. The effect was present in the dark, and illumination stimulated it only marginally. Low (less than 100 micromol/L) Ca2+ concentrations induced the formation of chlamydospores. Sr2+ could substitute Ca2+ in conidiogenesis with lower efficiency (almost 2 orders of magnitude), while the efficiency of Mg2+, Mn2+, or Ba2+ was lower by almost 3 orders of magnitude. Our results demonstrate that mycelial Ca2+ homeostasis has powerful effects on the conidiation and mycelial morphogenesis in T. viride, and they suggest that there is an additional mechanism of conidiation in addition to those induced by light and starvation. PMID- 18389002 TI - Short-term temporal dynamics of yeast abundance on the tall fescue phylloplane. AB - Six replicate trials were conducted to determine the short-term temporal dynamics and the effects of foliar applications of nutrients on the phylloplane yeast community of tall fescue (Festuca arundinacea Schreb.). In each trial, 2% sucrose + 0.5% yeast extract solution or sterile deionized water (control) was applied to the experiment plots. Twelve hours post-treatment (at 0600 hours), leaf samples were collected and yeast colony-forming units (cfu) were enumerated by dilution plating. This process was repeated at 1200, 1800, and 2400 hours in each trial. Significant differences were observed between the number of yeast cfu and the time at which the samples were collected. On average, the number of yeast cfu recovered was significantly less at 1800 hours and significantly greatest at 2400 hours when compared with all other sampling times. Averaged over all time intervals, we observed a trend of increased yeast abundance in turf treated with the nutrient solution compared with control treatments. In a separate investigation, atmospheric yeast abundance above the canopy of tall fescue was assessed in the morning (0900) and in the afternoon (1500) using a Thermo Andersen single stage viable particle sampler. In 5 of the 6 trials of this experiment, atmospheric yeast abundance was significantly greater in the morning than in the afternoon. Results suggest the following colonization model: phylloplane yeasts on tall fescue reproduce during the late evening and early morning, stabilize during the late morning and early afternoon through exchange of immigrants and emigrants, and decline during the late afternoon and (or) early evening. PMID- 18389003 TI - Pretreatment of lignocellulosic material with fungi capable of higher lignin degradation and lower carbohydrate degradation improves substrate acid hydrolysis and the eventual conversion to ethanol. AB - Phanerochaete chrysosporium, Pycnoporus cinnabarinus,and fungal isolates RCK-1 and RCK-3 were tested for their lignin degradation abilities when grown on wheat straw (WS) and Prosopis juliflora (PJ) under solid-state cultivation conditions. Fungal isolate RCK-1 degraded more lignin in WS (12.26% and 22.64%) and PJ (19.30% and 21.97%) and less holocellulose in WS (6.27% and 9.39%) and PJ (3.01% and 4.58%) after 10 and 20 days, respectively, than other fungi tested. Phanerochaete chrysosporium caused higher substrate mass loss and degraded more of holocellulosic content (WS: 55.67%; PJ: 48.89%) than lignin (WS: 18.89%; PJ: 20.20%) after 20 days. The fungal pretreatment of WS and PJ with a high-lignin degrading and low-holocellulose-degrading fungus (fungal isolate RCK-1) for 10 days resulted in (i) reduction in acid load for hydrolysis of structural polysaccharides (from 3.5% to 2.5% in WS and from 4.5% to 2.5% in PJ), (ii) an increase in the release of fermentable sugars (from 30.27 to 40.82 g L(-1) in WS and from 18.18 to 26.00 g L(-1) in PJ), and (iii) a reduction in fermentation inhibitors (total phenolics) in acid hydrolysate of WS (from 1.31 to 0.63 g L( 1)) and PJ (from 2.05 to 0.80 g L(-1)). Ethanol yield and volumetric productivity from RCK-1-treated WS (0.48 g g(-1) and 0.54 g L(-1) h(-1), respectively) and PJ (0.46 g g(-1) and 0.33 g L(-1) h(-1), respectively) were higher than untreated WS (0.36 g g(-1) and 0.30 g L(-1) h(-1), respectively) and untreated PJ (0.42 g g( 1) and 0.21 g L(-1) h(-1), respectively). PMID- 18389004 TI - Properties of an alginate-degrading Flavobacterium sp. strain LXA isolated from rotting algae from coastal China. AB - A novel bacterium exhibiting alginolytic activity was isolated from rotten algae. The alginate-degrading activity was detected in the culture supernatant by measuring the decrease in alginate viscosity or the increase in reducing sugars. Basic characterization showed that it was gram negative, rod shaped, yellow pigmented, and positive for oxidase and catalase, with a DNA G+C content of 35.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that this strain is related to members of the genus Flavobacterium. Sequence similarity values with their nearest phylogenetic neighbours ranged from 95.9% to 96.7%. Genotypic results, together with phenotypic characteristics, differentiated this species from related Flavobacterium organisms with validly published names, which suggests that the organism should be a new species of the genus Flavobacterium tentatively named as Flavobacterium sp. strain LXA. PMID- 18389005 TI - Identification of novel horA-harbouring bacteria capable of spoiling beer. AB - An ATP-binding cassette (ABC) multi-drug resistance (MDR) gene was found in 4 Gram-positive bacterial isolates of environmental origin and found capable of spoiling beer. The bacteria isolated were Bacillus cereus, Bacillus licheniformis, Paenibacillus humicus, and Staphylococcus epidermidis; all of which were previously unappreciated as beer-spoilage bacteria. The MDR gene found in these bacteria has less than 37% similarity to known ABC MDR proteins described for Bacillus and Staphylococcus, and this is the first finding of an ABC MDR gene in the genus Paenibacillus. The sequenced region of the gene was translated and compared phylogenetically with the closest GenBank matches of the respective species and the closest GenBank matches overall. The ABC MDR proteins from these isolates were found to cluster among known sequences of HorA, sharing 99.5% identity within the sequenced region. In the beer-spoilage-associated genera Lactobacillus and Pediococcus, the presence of the MDR gene horA correlates with the ability to grow in beer. As the unique horA-harbouring isolates described here are capable of growing in beer, it is likely that the presence of the horA gene likewise confers hop resistance to these organisms. PMID- 18389007 TI - Semiselective isolation of the ethanol-imbibing sooty mould Baudoinia of distillery aging warehouses. AB - Baudoinia compniacensis is a darkly pigmented microfungus that grows conspicuously on environmental surfaces around warehouses where alcoholic spirits are stored in wooden casks. This fungus has long been ignored because its primary isolation is very difficult. The present study describes a new semiselective isolation medium for this fungus based on the use of ethanol as a sole carbon source and low levels of nitrogen and trace elements. PMID- 18389006 TI - Occurrence of herpes simplex virus 1 and three periodontal bacteria in patients with chronic periodontitis and necrotic pulp. AB - Viral and bacterial associations appear to be implicated in the development of periodontal infections. Little information is available describing the periodontopathic agents in root canals with necrotic pulp. In this study, the occurrence and the combinations among herpes simplex virus type 1 (HSV-1) and Dialister pneumosintes, Tannerella forsythia, and Treponema denticola in patients with chronic periodontitis and necrotic pulp were evaluated. Clinical samples from healthy subjects and patients with periodontal or pulp infections were analyzed using a nested polymerase chain reaction PCR to detect HSV and PCR to detect the 3 periodontal bacteria. The presence of Tannerella forsythia and Treponema denticola was observed in healthy, periodontitis, and necrotic pulp patients. HSV was observed in periodontitis and necrotic pulp patients, and no healthy subject harbored D. pneumosintes or HSV. The occurrence of Tannerella forsythia was not statistically significant in patients with necrotic pulp (P = 0.704). Periodontal bacteria were observed varying from 10.3% to 20.7% in periodontitis and necrotic pulp patients. The presence of Treponema denticola - HSV association was predominant in patients showing necrotic pulp (24.1%); however, HSV alone was observed in one patient with periodontitis and in another patient with necrotic pulp. The presence of double association among bacteria or bacteria - HSV could indicate a role in both periodontitis and necrotic pulp, and Tannerella forsythia - Treponema denticola - HSV and Tannerella forsythia - D. pneumosintes - Treponema denticola - HSV associations might be important in periodontitis. PMID- 18389008 TI - Pyomelanin is produced by Shewanella algae BrY and affected by exogenous iron. AB - Melanin production by Shewanella algae BrY occurred during late- and (or) post exponential growth in lactate basal salts liquid medium supplemented with tyrosine or phenylalanine. The antioxidant ascorbate inhibited melanin production but not production of the melanin precursor homogentisic acid. In the absence of ascorbate, melanin production was inhibited by the 4-hydroxyphenylpyruvate dioxygenase inhibitor sulcotrione and by concentrations of Fe >or= 0.38 mmol L( 1). These data support the hypothesis that pigment production by S. algae BrY was a result of the conversion of tyrosine or phenylalanine to homogentisic acid, which was excreted, auto-oxidized, and self-polymerized to form pyomelanin. Pyomelanin production by S. algae BrY may play an important role in the biogeochemical cycling of Fe in the environment. PMID- 18389009 TI - Nicotinamide and 1-methylnicotinamide reduce homocysteine neurotoxicity in primary cultures of rat cerebellar granule cells. AB - Nicotinamide is an important cofactor in many metabolic pathways and a known neuroprotective substance, while its methylated product, 1-methylnicotinamide, is a suspected neurotoxin. Homocysteine is a risk factor in Alzheimer's disease and neurodegeneration, causing inhibition of methylation processes and inducing excitotoxicity. In this study, using primary cultures of rat cerebellar granule cells and propidium iodide staining, we investigated the neurotoxicity of nicotinamide and 1-methylnicotinamide, and their neuroprotective potential in acute and sub-acute homocysteine neurotoxicity. Our results demonstrated that nicotinamide and 1-methylnicotinamide applied for 24 h to cultures at concentrations of up to 25 mM had no effect on neuronal viability. Moreover, nicotinamide at concentrations of 5-20 mM and 1-methylnicotinamide at 1-10 mM applied to cells 24 h before, and for 24 h after an acute 30 min application of 25 mM D,L homocysteine, reduced neuronal damage. 1-Methylnicotinamide at concentrations of 250 and 500 muM showed neuroprotective activity during a sub acute 24-h exposure to 2.5 mM D,L-homocysteine, while 5 and 25 mM nicotinamide also evoked neuroprotection. These findings do not support suggestions that 1 methylnicotinamide may act as an endogenous neurotoxic agent; rather, they indicate the neuroprotective ability of nicotinamide and 1-methylnicotinamide in homocysteine neurotoxicity. The exact mechanisms of this neuroprotection are unclear and require further investigation. PMID- 18389010 TI - Modulation of Ca2+ channel current by mu opioid receptors in prefrontal cortex pyramidal neurons in rats. AB - Our work assesses the effects of mu opioid receptor activation on high-threshold Ca2+/Ba2+ currents in freshly dispersed pyramidal neurons of the medial prefrontal cortex in rats. Application of the specific mu receptor agonist (D Ala2+, N-Me-Phe4+, Gly5+-ol)-enkephalin (DAMGO) at 1 microM decreased Ca2+ current amplitudes from 0.72 to 0.49 nA. The effect was abolished by naloxone and omega-Conotoxin GVIA. Inhibition was not abolished by strong depolarisation of the cell membrane. In addition, a macroscopic Ba2+ current recorded in cell attached configuration was inhibited when DAMGO was applied outside the patch pipette. An adenylyl cyclase inhibitor (SQ 22536) and a protein kinase A inhibitor (H-89) decreased Ca2+ current amplitude. Moreover, the inhibitory effect of mu opioid receptors on Ca2+ currents required the activation of protein kinase A. We conclude that activation of mu opioid receptors in medial prefrontal cortex pyramidal neurons inhibits N type Ca2+ channel currents, and that protein kinase A is involved in this transduction pathway. PMID- 18389011 TI - Inhibition of neuronal nitric oxide synthase prevents iron-induced cerebellar Purkinje cell loss in the rat. AB - Iron plays an important role in maintaining normal brain function. However, in many neurodegenerative diseases abnormal iron accumulation in specific brain regions has been consistently reported. In this study, we investigated the neurotoxic effect of the intracerebroventricularly injected iron on the cerebellar Purkinje cells in the rat and the role of nitric oxide (NO) in this process. The role of NO in rats administered iron (FeCl36H2O) was examined with the use of a donor of NO, L-arginine (L-Arg) and a central selective inhibitor of NO synthase, 7-nitroindazole (7-NI). For this reason, rats were divided into 5 groups: control, iron-injected, iron plus L-Arg, iron plus 7-NI, and iron plus L Arg plus 7-NI. Means (value +/- standard deviation) of the total numbers of Purkinje cells in the cerebellum were estimated as 337 +/- 23, 209 +/- 16, 167 +/ 19, 305 +/- 26, and 265 +/- 14 thousands in the control, iron, iron plus L-Arg, iron plus 7-NI, and iron plus L-Arg plus 7-NI groups, respectively. Iron treatment alone and the combination of iron and L-Arg caused a significant reduction in the total number of cerebellar Purkinje cells. Therefore, L-Arg increased the Purkinje cell loss induced by treatment with iron. These data show that inhibition of the neuronal NOS by 7-NI can prevent some of the deleterious effects of iron on cerebellar Purkinje cells. Presence of L-arginine decreased the neuroprotective effect of 7-NI. PMID- 18389012 TI - Modulation of radiation-induced biochemical changes in cerebrum of Swiss albino mice by Grewia Asiatica. AB - The present study evaluates the possible radioprotective effect of Grewia asiatica fruit (rich in anthocyanin, carotenes, vitamin C, etc.) pulp extract (GAE) on cerebrum of Swiss albino mice exposed to 5 Gy gamma radiation. For this, healthy mice from an inbred colony were divided into four groups: (1) Control (vehicle treated) (2) GAE treated - mice in this group were orally supplemented with GAE (700 m/kg. b.w./day) once daily for fifteen consecutive days, (3) Vehicle treated irradiated mice, and (4) GAE + Irradiated - Mice in this group received distilled water orally equivalent to GAE (700 m/kg. b.w/day) for fifteen days consecutively. Mice were sacrificed at various intervals viz. 1-30 days. Radiation-induced augmentation in the levels of lipid peroxidation of mice cerebrum was significantly ameliorated by GAE pretreatment. Radiation-induced depletion in the level of glutathione and protein was prevented significantly by GAE administration. PMID- 18389013 TI - Tubulovesicular structures are present in brains of hamsters infected with the Echigo-1 strain of Creutzfeldt-Jakob disease agent. AB - Tubulovesicular structures (particles; TVS) are virion-like particles 25-30 nm in diameter found by thin-section electron microscopy in brains of all prion diseases including scrapie, Creutzfeldt-Jakob disease (CJD) fatal familial insomnia (FFI) and Gerstmann-Straussler-Scheineker disease (GSS) as well as in cell cultures infected with TSE agents. TVS are regarded as a disease-specific ultrastructural marker for TSEs and, by those not completely satisfied with the prion hypothesis, they are even considered to be a possible candidate for the infectious TSE agent itself. A caveat regarding that interpretation stemmed from previous failures to find TVS by electron microscopic studies of tissues from animals infected with the Echigo-1 strain of CJD agent. We now report detecting TVS in brains of hamsters infected with that strain of CJD agent, albeit with a very low frequency. PMID- 18389014 TI - Differential endocrine response in rams to intracerebroventricular infusion of genistein. AB - The intracerebroventricular infusions of genistein (total 40 mug) were made in male sheep (November) to test its influence on melatonin, growth hormone (GH) and luteinizing hormone (LH) secretion. The analysis of the results encompassed 3 similar periods: before the infusion (afternoon hours) the first (evening hours) and the second (night hours) halves of the treatment. The night plasma concentration of melatonin in genistein-infused rams was significantly lower than that noted during the respective period in vehicle-infused rams. Plasma GH concentration increased significantly in both vehicle- and genistein-infused rams during the night hours, as compared with the concentrations noted during the afternoon and evening, however, genistein significantly stimulated the amplitude of GH pulses in these latter. The LH concentration was significantly lower during the second part of genistein treatment, than in vehicle-infused rams. The frequency and amplitude of LH pulses clearly tended to decrease following genistein infusion. In conclusion, genistein, acting at the central nervous system level in sexually active rams is able to reduce the secretion of melatonin and LH and has also a slight stimulatory effect on the amplitude of GH pulses. PMID- 18389015 TI - Sway ratio - a new measure for quantifying postural stability. AB - In the search of a reliable postural stability index, two sway time series: the center-of-mass (COM) and the center-of-foot pressure (COP) were recorded simultaneously in elderly subjects standing quiet with eyes open and with eyes closed. From a battery of commonly use sway measures, only the anteroposterior COM and the COP path lengths proved their high sensitivity and discriminative power to the imposed vision conditions. Based upon these indices, a new measure - sway ratio (SR) - was computed, as the COP-to-COM path length ratio. The measure can easily distinguish vision vs. no vision in the elderly. The SR can be successfully accessed base upon the COP signal only. In contrast to traditional sway indices, the SR as a relative measure is insensitive to the length of sampled record and to the signal sampling frequency. Its magnitude can be interpreted as an average amount of balance controlling motor activity that coincides with a unit COM displacement. The SR is recommended as a reliable measure that allows for assessment of postural stability. PMID- 18389016 TI - Nonverbal deficits in explicit and implicit memory of Parkinson's disease patients. AB - This study examined verbal and nonverbal aspects of explicit and implicit memory in a sample of 19 Parkinson's disease (PD) patients and 21 control subjects. For implicit memory evaluation, we used a Mirror Reading (MR) task employing verbal material as well as a nonverbal Serial Reaction Time (SRT) task. For explicit memory measurement we applied a word pairs task (verbal) and pairs of a Japanese ideograms task (nonverbal). The PD patients displayed impairments in the nonverbal tasks only, namely, in the SRT task and the pairs of Japanese ideograms task. No correlation between Wisconsin Card Sorting Test (WCST) scores and the results of tasks in which PD patients displayed deficits (SRT and pairs of Japanese ideograms) were discovered. Interestingly, such a correlation was found in the case of MR and words pairs tasks, which did not distinguish PD patients from control group. PMID- 18389017 TI - Cognitive subtypes of dyslexia. AB - Different theories conceptualise dyslexia as either a phonological, attentional, auditory, magnocellular, or automatisation deficit. Such heterogeneity suggests the existence of yet unrecognised subtypes of dyslexics suffering from distinguishable deficits. The purpose of the study was to identify cognitive subtypes of dyslexia. Out of 642 children screened for reading ability 49 dyslexics and 48 controls were tested for phonological awareness, auditory discrimination, motion detection, visual attention, and rhythm imitation. A combined cluster and discriminant analysis approach revealed three clusters of dyslexics with different cognitive deficits. Compared to reading-unimpaired children cluster no. 1 had worse phonological awareness; cluster no. 2 had higher attentional costs; cluster no. 3 performed worse in the phonological, auditory, and magnocellular tasks. These results indicate that dyslexia may result from distinct cognitive impairments. As a consequence, prevention and remediation programmes should be specifically targeted for the individual child's deficit pattern. PMID- 18389018 TI - Behavioral manifestations of brain plasticity in blind and low-vision individuals. AB - Tactile sensitivity enhancement (TSE) observed in blind people is probably a result of intensified tactile training. Although many researchers consider TSE in the blind to be an example of use-dependent plasticity, it is unclear whether the effects of training (Braille reading) are specific, i.e. restricted to the trained function and hand, or if they are more general. To examine this issue further, blind Braille readers, low-vision subjects (Braille readers and non Braille readers) and sighted controls were tested in two tasks: a texture task resembling the Braille system and a dissimilar groove orientation task. Braille readers, both blind and those with low vision, performed better in both tasks than low-vision non-Braille readers or sighted controls. However, the difference was significant only for the blind (more experienced) Braille readers. In the groove orientation task, the positive influence of training was detectable irrespective of the hand used in the test, but in the coarse texture task this influence was limited to the hand trained in Braille. Thus, it appears that tactile training is of significance in TSE but its effects are, to a large extent, task- and hand-specific. PMID- 18389019 TI - Cortical evoked responses to magnetic stimulation of macaque's abdominal wall in sleep-wake cycle. AB - EEG and eye movements (magnetic search coil method) were recorded in sleep and wakefulness in a monkey (Macaca fascicularis) while the animal was sitting in a primate chair. Single pulse magnetic stimulation was applied to the monkey's abdominal wall using a circular coil and a Magstim 200 stimulator. Magnetic stimuli did not wake the sleeping animal, and being applied during slow wave sleep evoked clear responses in EEG with a latency of 80-100 ms. These responses disappeared during wakefulness and rapid eye movement sleep. Control experiments confirmed that these responses were not caused by the acoustic clicks produced by the magnetic coil. Results of this study further confirm that during sleep, signals from visceral organs reach the cortical areas which in wakefulness process exteroceptive sensory information. This observation indicates that magnetic stimulation may be a useful tool for researching neural connectivity reorganization within the sleep-wake cycle. PMID- 18389020 TI - Mismatch negativity in methamphetamine dependence: a pilot study. AB - The objective of this study was to verify hypothesised changes in event related potentials (visual mismatch negativity, vMMN) in 17 subjects dependent on methamphetamine (MAMP) compared to age and gender matched 17 healthy volunteers. We found a significant correlation between vMMN and duration of methamphetamine abuse (Spearman correlation coefficient r=0.54-0.78; P<.05). The positive correlation indicates drop of originally more negative response to deviant stimulus, what may indicate a pre-attentive processing enhancement in the first years of MAMP abuse with its decease later on. Accordingly, post-hoc analysis revealed significantly stronger vMMN in patients with length of MAMP abuse shorter than 5 years than in paired controls. There were no such differences in abusers with the length of abuse longer than 5 years. The results show that the visual processing on the pre-attentional level can be influenced by long-term MAMP abuse, what can be specifically assessed by vMMN. PMID- 18389021 TI - Transmission of information during Continuous Attention Test. AB - The Short-Time Directed Transfer Function (SDTF) is an estimator based on a multivariate autoregressive model which has proved to be successful in ERP experiments, e.g. those connected with motor action and its imagination. The aim of this study is the evaluation of the performance of SDTF in the cognitive experiment. We have applied SDTF for the estimation of the pattern of EEG signal transmissions during a Continuous Attention Test (CAT). Time-frequency patterns of propagation were estimated for two experimental conditions. Statistical procedures based on thin-plate spline model were used for estimation of significant changes in respect to the reference epoch. The repeatability of the results for a subject and across the subjects were investigated. The effect of prolonged transmission in the gamma band from the prefrontal electrodes found in all subjects was explained by the active inhibition in the case when a subject had to sustain from performing the action. PMID- 18389022 TI - The tubulovesicular structures - the ultrastructural hallmark for all prion diseases. AB - Tubulovesicular structures (particles - TVS) are the only ultrastructural marker for all prion diseases as seen by thin-section electron microscopy as opposed to "negative-staining" techniques. TVS are spheres or short rods of approximately 27 nm in diameter. That size of TVS is also the size of filter cut-off of infectivity as judged from the ultrafiltration studies and the size of the smallest infectious unit as recently estimated. TVS have been found in all naturally occurring and experimentally induced prion diseases, including variant Creutzfeldt-Jakob disease and human familial TSEs - fatal familial insomnia and Gerstmann-Straussler-Scheinker disease. In longitudinal studies, the number of neuronal processes containing TVS correlates roughly with the incubation period and with infectivity. Hence, they are readily found in hamsters infected with the 263K strain of scrapie but it is very difficult to find them in human TSEs where titer is lower. The composition of TVS is unknown but they are not composed of PrP. Their consistent presence in all TSEs suggests the unexplained role at least of TSE pathogenesis. PMID- 18389023 TI - [Editorial choices and publication profile]. PMID- 18389024 TI - [Abortion pill--a medication of women's choice]. PMID- 18389025 TI - [Medical progress with economic headache]. PMID- 18389026 TI - [Appropriate use of laboratory tests--medical aspects]. AB - BACKGROUND: There has been a large increase in the use and costs of laboratory tests during recent years. Several reports have indicated excessive and inappropriate use. The purpose of this study was to assess the use of public laboratory services within clinical chemistry in two Norwegian health regions. MATERIAL AND METHODS: Production statistics for 2004 were obtained through a questionnaire sent to all public clinical chemistry hospital laboratories in northern and western Norway. Additional detailed production statistics were obtained from Haukeland University Hospital for 2002-04. RESULTS: We observed differences in the absolute frequency of requested tests and a marked variation in relative ratios (ratio between related tests) between the laboratories in northern and western Norway. Data from Haukeland University Hospital showed a mean increase of 12% (range: -24-54%) in the number of ordered tests between 2002 04. INTERPRETATION: There are no known differences in morbidity between the northern and western health regions that can explain the observed variations in the use of laboratory tests. Our observations indicate a need for a thorough investigation of current utilisation of laboratory tests. Initiatives should be taken on a national basis to improve appropriate use. PMID- 18389027 TI - [More adequate use of laboratory services--economical aspects]. AB - BACKGROUND: There has been a substantial increase in reimbursement for outpatient laboratory services in recent years. This article gives an overview of the use of such laboratory services and discusses measures for improved efficiency. MATERIAL AND METHODS: We have analysed reimbursement to the specialist health care for the period 2002-04. RESULTS: In the period 2002-04 the reimbursement to public laboratories increased by 42%. There has been a substantial growth in all reimbursements groups, especially for "General analysis for molecular biological methods" (236%). Reimbursement to private laboratories have increased by 24%, similar to the increase in general clinical chemistry. There has been a trend towards using the expensive reimbursement groups more often, especially for public laboratories. There is substantial variation in the use of laboratory services between the health regions. DISCUSSION: Our results indicate that the growth and variation in the use of laboratory services cannot be explained by deteriorated health or more illness in the population. The variation indicates that the use of laboratory services is not optimal. Establishment of a national laboratory system within specialized health care may contribute to a more knowledge-based use of laboratory services. PMID- 18389028 TI - [Resection of symptomatic intraspinal meningiomas]. AB - BACKGROUND: Meningioma is one of the most common primary intraspinal tumours in adults. Most patients with this condition develop myelopathy or radiculopathy. The treatment of choice for these tumours is surgery, although few data are published on neurological outcome after surgery for intraspinal meningiomas. Patients are increasingly asking about the prognosis before they consent to surgery. This study aims at investigating operative morbidity and neurological outcome in a continuous surgical series in Norway. MATERIAL AND METHODS: The material comprised all 28 patients treated for histologically verified intraspinal meningioma at Rikshospitalet in the time period 1999 to 2003. All patients underwent new radiological and clinical assessments for study purposes. RESULTS AND INTERPRETATION: We found that 96.5% of the patients were neurologically improved or unchanged after surgery. There was no recurrence, nor mortality. These findings match earlier reports in the literature. Morbidity was at 18%, this was higher than in other studies due to our wide definition of surgical morbidity. We conclude that surgery can be performed with excellent outcome, whilst the morbidity and mortality remain low. PMID- 18389029 TI - [Use of cannabinoids in palliative medicine]. AB - BACKGROUND: Several countries use drugs containing cannabinoids for symptom management in patients that need palliative treatment. MATERIAL AND METHOD: Articles were retrieved through a literature search for randomized controlled studies of cannabinoids used to treat cancer-related symptoms such as pain, nausea and appetite. RESULTS: Evidence for clinical effect is limited. Some randomized studies have shown a treatment effect of cannabinoids in neuropathic pain. Cannabinoid-containing drugs can have a moderate anti-emetic effect and can stimulate appetite in some subgroups of patients. INTERPRETATION: Many questions about clinical efficacy and side effects have still not been answered for cannabinoid-containing drugs and there is not sufficient evidence available from relevant randomised controlled trials to base a decision on whether or not to permit such drugs in palliative care in Norway. It is however likely that drugs containing cannabinoids can help to control certain symptoms in some patients. PMID- 18389030 TI - [Maintenance treatment of chronic obstructive pulmonary disease]. AB - BACKGROUND: Treatment guidelines for chronic obstructive pulmonary disease (COPD) recommend that disease severity should be decisive for the choice of drug. MATERIAL AND METHODS: Medical records for patients with COPD were reviewed and an overview was made of the different drugs used for maintenance treatment. We examined the correspondence between drug classes used and guideline recommendations. RESULTS: Glucocorticosteroids and long-acting bronchodilators are used more frequently for maintenance treatment than anticholinergic drugs and short-acting bronchodilators. The severity of COPD does not influence the prescription of glucocorticosteroides and long-acting bronchodilators. INTERPRETATION: The physician's first choice of drug seems to continue as maintenance treatment. The results suggest that glucocorticosteroids are used more frequently than recommended by the COPD treatment guidelines. PMID- 18389031 TI - [Drugs to reduce transfusion needs during surgery]. AB - Blood components are a limited and expensive resource, and transfusions may cause serious side effects. Drug treatment is an option to reduce the need for transfusions related to surgery. Tranexamic acid reduces transfusion requirements after total hip and knee arthroplasty, and after various cardiac surgical procedures. Desmopressin does not reduce the need for transfusions after surgery in patients with normal preoperative hemostasis. Treatment with recombinant factor VIIa may be considered in patients with massive hemorrhage caused by blunt trauma, post-partum hemorrhage, cardiac surgery, and in uncontrolled bleeding in surgery in general. PMID- 18389032 TI - [The "idea polyclinic"--from research to society's benefit]. PMID- 18389033 TI - [The generic list--a drug-political tool]. PMID- 18389034 TI - [How can the use of laboratories be directed?]. PMID- 18389036 TI - [Mental retardation]. PMID- 18389037 TI - [Fat intake and cardiovascular health--are we wrongly informed?]. PMID- 18389040 TI - [Ultrasound--a good alternative to MR in shoulder problems]. PMID- 18389041 TI - [Individual food stocks as disaster planning]. PMID- 18389044 TI - [Nursing homes and medical treatment]. PMID- 18389046 TI - [Systematic ethical work in intensive care units is beneficial]. PMID- 18389047 TI - [End-of-life care--reports from Helsetilsynet]. PMID- 18389049 TI - [Women's health in Gambia]. PMID- 18389056 TI - A systematic assessment of MHC class II peptide binding predictions and evaluation of a consensus approach. AB - The identification of MHC class II restricted peptide epitopes is an important goal in immunological research. A number of computational tools have been developed for this purpose, but there is a lack of large-scale systematic evaluation of their performance. Herein, we used a comprehensive dataset consisting of more than 10,000 previously unpublished MHC-peptide binding affinities, 29 peptide/MHC crystal structures, and 664 peptides experimentally tested for CD4+ T cell responses to systematically evaluate the performances of publicly available MHC class II binding prediction tools. While in selected instances the best tools were associated with AUC values up to 0.86, in general, class II predictions did not perform as well as historically noted for class I predictions. It appears that the ability of MHC class II molecules to bind variable length peptides, which requires the correct assignment of peptide binding cores, is a critical factor limiting the performance of existing prediction tools. To improve performance, we implemented a consensus prediction approach that combines methods with top performances. We show that this consensus approach achieved best overall performance. Finally, we make the large datasets used publicly available as a benchmark to facilitate further development of MHC class II binding peptide prediction methods. PMID- 18389057 TI - The statistical neuroanatomy of frontal networks in the macaque. AB - We were interested in gaining insight into the functional properties of frontal networks based upon their anatomical inputs. We took a neuroinformatics approach, carrying out maximum likelihood hierarchical cluster analysis on 25 frontal cortical areas based upon their anatomical connections, with 68 input areas representing exterosensory, chemosensory, motor, limbic, and other frontal inputs. The analysis revealed a set of statistically robust clusters. We used these clusters to divide the frontal areas into 5 groups, including ventral lateral, ventral-medial, dorsal-medial, dorsal-lateral, and caudal-orbital groups. Each of these groups was defined by a unique set of inputs. This organization provides insight into the differential roles of each group of areas and suggests a gradient by which orbital and ventral-medial areas may be responsible for decision-making processes based on emotion and primary reinforcers, and lateral frontal areas are more involved in integrating affective and rational information into a common framework. PMID- 18389058 TI - Deciphering interplay between Salmonella invasion effectors. AB - Bacterial pathogens have evolved a specialized type III secretion system (T3SS) to translocate virulence effector proteins directly into eukaryotic target cells. Salmonellae deploy effectors that trigger localized actin reorganization to force their own entry into non-phagocytic host cells. Six effectors (SipC, SipA, SopE/2, SopB, SptP) can individually manipulate actin dynamics at the plasma membrane, which acts as a 'signaling hub' during Salmonella invasion. The extent of crosstalk between these spatially coincident effectors remains unknown. Here we describe trans and cisbinary entry effector interplay (BENEFIT) screens that systematically examine functional associations between effectors following their delivery into the host cell. The results reveal extensive ordered synergistic and antagonistic relationships and their relative potency, and illuminate an unexpectedly sophisticated signaling network evolved through longstanding pathogen-host interaction. PMID- 18389059 TI - Evolution and control of imprinted FWA genes in the genus Arabidopsis. AB - A central question in genomic imprinting is how a specific sequence is recognized as the target for epigenetic marking. In both mammals and plants, imprinted genes are often associated with tandem repeats and transposon-related sequences, but the role of these elements in epigenetic gene silencing remains elusive. FWA is an imprinted gene in Arabidopsis thaliana expressed specifically in the female gametophyte and endosperm. Tissue-specific and imprinted expression of FWA depends on DNA methylation in the FWA promoter, which is comprised of two direct repeats containing a sequence related to a SINE retroelement. Methylation of this element causes epigenetic silencing, but it is not known whether the methylation is targeted to the SINE-related sequence itself or the direct repeat structure is also necessary. Here we show that the repeat structure in the FWA promoter is highly diverse in species within the genus Arabidopsis. Four independent tandem repeat formation events were found in three closely related species. Another related species, A. halleri, did not have a tandem repeat in the FWA promoter. Unexpectedly, even in this species, FWA expression was imprinted and the FWA promoter was methylated. In addition, our expression analysis of FWA gene in vegetative tissues revealed high frequency of intra-specific variation in the expression level. In conclusion, we show that the tandem repeat structure is dispensable for the epigenetic silencing of the FWA gene. Rather, SINE-related sequence is sufficient for imprinting, vegetative silencing, and targeting of DNA methylation. Frequent independent tandem repeat formation events in the FWA promoter led us to propose that they may be a consequence, rather than cause, of the epigenetic control. The possible significance of epigenetic variation in reproductive strategies during evolution is also discussed. PMID- 18389060 TI - A novel secretion pathway of Salmonella enterica acts as an antivirulence modulator during salmonellosis. AB - Salmonella spp. are Gram-negative enteropathogenic bacteria that infect a variety of vertebrate hosts. Like any other living organism, protein secretion is a fundamental process essential for various aspects of Salmonella biology. Herein we report the identification and characterization of a horizontally acquired, autonomous and previously unreported secretion pathway. In Salmonella enterica serovar Typhimurium, this novel secretion pathway is encoded by STM1669 and STM1668, designated zirT and zirS, respectively. We show that ZirT is localized to the bacterial outer membrane, expected to adopt a compact beta-barrel conformation, and functions as a translocator for ZirS. ZirS is an exoprotein, which is secreted into the extracellular environment in a ZirT-dependent manner. The ZirTS secretion pathway was found to share several important features with two-partner secretion (TPS) systems and members of the intimin/invasin family of adhesions. We show that zirTS expression is affected by zinc; and that in vivo, induction of zirT occurs distinctively in Salmonella colonizing the small intestine, but not in systemic sites. Additionally, strong expression of zirT takes place in Salmonella shed in fecal pellets during acute and persistent infections of mice. Inactivation of ZirTS results in a hypervirulence phenotype of Salmonella during oral infection of mice. Cumulatively, these results indicate that the ZirTS pathway plays a unique role as an antivirulence modulator during systemic disease and is involved in fine-tuning a host-pathogen balance during salmonellosis. PMID- 18389061 TI - Tobacco mosaic virus movement protein enhances the spread of RNA silencing. AB - Eukaryotic cells restrain the activity of foreign genetic elements, including viruses, through RNA silencing. Although viruses encode suppressors of silencing to support their propagation, viruses may also exploit silencing to regulate host gene expression or to control the level of their accumulation and thus to reduce damage to the host. RNA silencing in plants propagates from cell to cell and systemically via a sequence-specific signal. Since the signal spreads between cells through plasmodesmata like the viruses themselves, virus-encoded plasmodesmata-manipulating movement proteins (MP) may have a central role in compatible virus:host interactions by suppressing or enhancing the spread of the signal. Here, we have addressed the propagation of GFP silencing in the presence and absence of MP and MP mutants. We show that the protein enhances the spread of silencing. Small RNA analysis indicates that MP does not enhance the silencing pathway but rather enhances the transport of the signal through plasmodesmata. The ability to enhance the spread of silencing is maintained by certain MP mutants that can move between cells but which have defects in subcellular localization and do not support the spread of viral RNA. Using MP expressing and non-expressing virus mutants with a disabled silencing suppressing function, we provide evidence indicating that viral MP contributes to anti-viral silencing during infection. Our results suggest a role of MP in controlling virus propagation in the infected host by supporting the spread of silencing signal. This activity of MP involves only a subset of its properties implicated in the spread of viral RNA. PMID- 18389062 TI - The MHV68 M2 protein drives IL-10 dependent B cell proliferation and differentiation. AB - Murine gammaherpesvirus 68 (MHV68) establishes long-term latency in memory B cells similar to the human gammaherpesvirus Epstein Barr Virus (EBV). EBV encodes an interleukin-10 (IL-10) homolog and modulates cellular IL-10 expression; however, the role of IL-10 in the establishment and/or maintenance of chronic EBV infection remains unclear. Notably, MHV68 does not encode an IL-10 homolog, but virus infection has been shown to result in elevated serum IL-10 levels in wild type mice, and IL-10 deficiency results in decreased establishment of virus latency. Here we show that a unique MHV68 latency-associated gene product, the M2 protein, is required for the elevated serum IL-10 levels observed at 2 weeks post infection. Furthermore, M2 protein expression in primary murine B cells drives high level IL-10 expression along with increased secretion of IL-2, IL-6, and MIP 1alpha. M2 expression was also shown to significantly augment LPS driven survival and proliferation of primary murine B cells. The latter was dependent on IL-10 expression as demonstrated by the failure of IL10-/- B cells to proliferate in response to M2 protein expression and rescue of M2-associated proliferation by addition of recombinant murine IL-10. M2 protein expression in primary B cells also led to upregulated surface expression of the high affinity IL-2 receptor (CD25) and the activation marker GL7, along with down-regulated surface expression of B220, MHC II, and sIgD. The cells retained CD19 and sIgG expression, suggesting differentiation to a pre-plasma memory B cell phenotype. These observations are consistent with previous analyses of M2-null MHV68 mutants that have suggested a role for the M2 protein in expansion and differentiation of MHV68 latently infected B cells-perhaps facilitating the establishment of virus latency in memory B cells. Thus, while the M2 protein is unique to MHV68, analysis of M2 function has revealed an important role for IL-10 in MHV68 pathogenesis-identifying a strategy that appears to be conserved between at least EBV and MHV68. PMID- 18389064 TI - Genome-wide prediction of SH2 domain targets using structural information and the FoldX algorithm. AB - Current experiments likely cover only a fraction of all protein-protein interactions. Here, we developed a method to predict SH2-mediated protein-protein interactions using the structure of SH2-phosphopeptide complexes and the FoldX algorithm. We show that our approach performs similarly to experimentally derived consensus sequences and substitution matrices at predicting known in vitro and in vivo targets of SH2 domains. We use our method to provide a set of high confidence interactions for human SH2 domains with known structure filtered on secondary structure and phosphorylation state. We validated the predictions using literature-derived SH2 interactions and a probabilistic score obtained from a naive Bayes integration of information on coexpression, conservation of the interaction in other species, shared interaction partners, and functions. We show how our predictions lead to a new hypothesis for the role of SH2 domains in signaling. PMID- 18389063 TI - Stimulation of chitin synthesis rescues Candida albicans from echinocandins. AB - Echinocandins are a new generation of novel antifungal agent that inhibit cell wall beta(1,3)-glucan synthesis and are normally cidal for the human pathogen Candida albicans. Treatment of C. albicans with low levels of echinocandins stimulated chitin synthase (CHS) gene expression, increased Chs activity, elevated chitin content and reduced efficacy of these drugs. Elevation of chitin synthesis was mediated via the PKC, HOG, and Ca(2+)-calcineurin signalling pathways. Stimulation of Chs2p and Chs8p by activators of these pathways enabled cells to survive otherwise lethal concentrations of echinocandins, even in the absence of Chs3p and the normally essential Chs1p, which synthesize the chitinous septal ring and primary septum of the fungus. Under such conditions, a novel proximally offset septum was synthesized that restored the capacity for cell division, sustained the viability of the cell, and abrogated morphological and growth defects associated with echinocandin treatment and the chs mutations. These findings anticipate potential resistance mechanisms to echinocandins. However, echinocandins and chitin synthase inhibitors synergized strongly, highlighting the potential for combination therapies with greatly enhanced cidal activity. PMID- 18389065 TI - The nature of the chemical bond in linear three-body systems: from i3- to mixed chalcogen/halogen and trichalcogen moieties. AB - The 3 centre-4 electrons (3c-4e) and the donor/acceptor or charge-transfer models for the description of the chemical bond in linear three-body systems, such as I(3) (-) and related electron-rich (22 shell electrons) systems, are comparatively discussed on the grounds of structural data from a search of the Cambridge Structural Database (CSD). Both models account for a total bond order of 1 in these systems, and while the former fits better symmetric systems, the latter describes better strongly asymmetric situations. The 3c-4e MO scheme shows that any linear system formed by three aligned closed-shell species (24 shell electrons overall) has reason to exist provided that two electrons are removed from it to afford a 22 shell electrons three-body system: all combinations of three closed-shell halides and/or chalcogenides are considered here. A survey of the literature shows that most of these three-body systems exist. With some exceptions, their structural features vary continuously from the symmetric situation showing two equal bonds to very asymmetric situations in which one bond approaches to the value corresponding to a single bond and the second one to the sum of the van der Waals radii of the involved atoms. This indicates that the potential energy surface of these three-body systems is fairly flat, and that the chemical surrounding of the chalcogen/halogen atoms can play an important role in freezing different structural situations; this is well documented for the I(3) ( ) anion. The existence of correlations between the two bond distances and more importantly the linearity observed for all these systems, independently on the degree of their asymmetry, support the state of hypervalency of the central atom. PMID- 18389066 TI - Dimensionality and dynamics in the behavior of C. elegans. AB - A major challenge in analyzing animal behavior is to discover some underlying simplicity in complex motor actions. Here, we show that the space of shapes adopted by the nematode Caenorhabditis elegans is low dimensional, with just four dimensions accounting for 95% of the shape variance. These dimensions provide a quantitative description of worm behavior, and we partially reconstruct "equations of motion" for the dynamics in this space. These dynamics have multiple attractors, and we find that the worm visits these in a rapid and almost completely deterministic response to weak thermal stimuli. Stimulus-dependent correlations among the different modes suggest that one can generate more reliable behaviors by synchronizing stimuli to the state of the worm in shape space. We confirm this prediction, effectively "steering" the worm in real time. PMID- 18389067 TI - An empirical bayesian method for detecting differentially expressed genes using EST data. AB - Detection of differentially expressed genes from expressed sequence tags (ESTs) data has received much attention. An empirical Bayesian method is introduced in which gene expression patterns are estimated and used to define detection statistics. Significantly differentially expressed genes can be declared given detection statistics. Simulation is done to evaluate the performance of proposed method. Two real applications are studied. PMID- 18389068 TI - Feasibility Study and Design of a Wearable System-on-a-Chip Pulse Radar for Contactless Cardiopulmonary Monitoring. AB - A new system-on-a-chip radar sensor for next-generation wearable wireless interface applied to the human health care and safeguard is presented. The system overview is provided and the feasibility study of the radar sensor is presented. In detail, the overall system consists of a radar sensor for detecting the heart and breath rates and a low-power IEEE 802.15.4 ZigBee radio interface, which provides a wireless data link with remote data acquisition and control units. In particular, the pulse radar exploits 3.1-10.6 GHz ultra-wideband signals which allow a significant reduction of the transceiver complexity and then of its power consumption. The operating principle of the radar for the cardiopulmonary monitoring is highlighted and the results of the system analysis are reported. Moreover, the results obtained from the building-blocks design, the channel measurement, and the ultra-wideband antenna realization are reported. PMID- 18389069 TI - Modified vertical rectus abdominis musculocutaneous flap for limb salvage procedures in proximal lower limb musculoskeletal sarcomas. AB - Introduction and aim. Management of complicated wounds after tumor extipiration of pelvic and proximal lower limb musculoskeletal sarcoma represents an essential component in the outcome of these patients. The authors present modified vertical rectus abdominis musculocutaneous (VRAM) flap techniques to reconstruct extensive defects after debridment of these complicated wounds. Material and Methods. Over a period of 4 years (2002-2005), 5 men and 2 women were managed. Median age was 21 years (range 15-49). The patients were managed for complicated lower trunk, groin, and upper thigh wounds after resection of three pelvic chondrosarcomas as well as two pelvic and two proximal femur osteosarcomas. The modifications included a VRAM flap with lateral and tongue-like extension design of the skin paddle (5 cases) or a delayed extended VRAM flap (2 cases). Results. All flaps showed complete survival and healing with no ischemic events providing stable coverage. All patients were ambulant with good limb functions in terms of walking and gait after adequate rehabilitation, 2 needed support with crutches. Conclusion. The modified VRAM flaps offer reliable reconstructive tools for coverage of complex groin and thigh defects by providing larger well-vascularized soft tissue with acceptable donor site. PMID- 18389071 TI - PPARs/RXRs in Cardiovascular Physiology and Disease. PMID- 18389072 TI - Metal based drugs restyled and resumed. PMID- 18389073 TI - A robotic voice simulator and the interactive training for hearing-impaired people. AB - A talking and singing robot which adaptively learns the vocalization skill by means of an auditory feedback learning algorithm is being developed. The robot consists of motor-controlled vocal organs such as vocal cords, a vocal tract and a nasal cavity to generate a natural voice imitating a human vocalization. In this study, the robot is applied to the training system of speech articulation for the hearing-impaired, because the robot is able to reproduce their vocalization and to teach them how it is to be improved to generate clear speech. The paper briefly introduces the mechanical construction of the robot and how it autonomously acquires the vocalization skill in the auditory feedback learning by listening to human speech. Then the training system is described, together with the evaluation of the speech training by auditory impaired people. PMID- 18389070 TI - Regulation of FoxP3 regulatory T cells and Th17 cells by retinoids. AB - Vitamin A has both positive and negative regulatory functions in the immune system. While vitamin A is required for normal formation of immune cells and epithelial cell barriers, vitamin A deficiency can lead to increased inflammatory responses and tissue damage. The mechanism with which vitamin A and its metabolites such as retinoids negatively regulate inflammatory responses has not been clearly defined. Recently, it has been established that retinoids promote the generation of immune-suppressive FoxP3+ regulatory T cells while they suppress the T cell differentiation into inflammatory Th17 cells in the periphery such as intestine. These novel functions of retinoids provide a potentially important immune regulatory mechanism. In this review, we discuss the functions of retinoids in the development of the FoxP3+ cells and Th17 cells, the phenotype and functions of retinoid-induced FoxP3+ T cells, and the impact of retinoid induced FoxP3+ T cells on the immune tolerance. PMID- 18389074 TI - Online preconcentration and determination of trace amounts of zinc in nature waters. AB - A simple, sensitive, reliable and flexible flow injection spectrophotometric method is proposed for on-line preconcentration and determination of trace amounts of zinc in water. At the presence of Tween-80 in pH 9.3 buffer solutions, the shade of color of Zn (II)-PAN complex is in a linear relation to the zinc amount at the point of the maximum absorption peak of 560 nm. The optimal experimental conditions, including reaction conditions and preconcentration conditions, had been obtained. The linear range of the proposed method was between 2.0 and 360 mug L(-1) and the detection limit was 0.42 mug L(-1). The relative standard deviation was 3.55% and 2.14% for 5.0 mug L(-1) and 50 mug L( 1) of zinc standard solution (n = 8). The method had been successfully applied to zinc determination in water samples and the analytical results were satisfactory. PMID- 18389076 TI - Influence of C-peptide on glucose utilisation. AB - During the recent years, multiple studies demonstrated that C-peptide is not an inert peptide, but exerts important physiological effects. C-peptide binds to cell membranes, stimulates the Na,K-ATPase and the endothelial nitric oxide (NO) synthase. Moreover, there is evidence that C-peptide decreases glomerular hyperfiltration and increases glucose utilisation. Nevertheless, there is still limited knowledge concerning mechanisms leading to an increased glucose utilisation either in rats or in humans. The aim of this paper is to give an overview over the published studies regarding C-peptide and glucose metabolism from in vitro studies to longer lasting studies in humans. PMID- 18389075 TI - The NF-kappaB inhibitor curcumin blocks sepsis-induced muscle proteolysis. AB - We tested the hypothesis that treatment of rats with curcumin prevents sepsis induced muscle protein degradation. In addition, we determined the influence of curcumin on different proteolytic pathways that are activated in septic muscle (i.e., ubiquitin-proteasome-, calpain-, and cathepsin L-dependent proteolysis) and examined the role of NF-kappaB and p38/MAP kinase inactivation in curcumin induced inhibition of muscle protein breakdown. Rats were made septic by cecal ligation and puncture or were sham-operated. Groups of rats were treated with three intraperitoneal doses (600 mg/kg) of curcumin or corresponding volumes of solvent. Protein breakdown rates were measured as release of tyrosine from incubated extensor digitorum longus muscles. Treatment with curcumin prevented sepsis-induced increase in muscle protein breakdown. Surprisingly, the upregulated expression of the ubiquitin ligases atrogin-1 and MuRF1 was not influenced by curcumin. When muscles from septic rats were treated with curcumin in vitro, proteasome-, calpain-, and cathepsin L-dependent protein breakdown rates were reduced, and nuclear NF-kappaB/p65 expression and activity as well as levels of phosphorylated (activated) p38 were decreased. Results suggest that sepsis-induced muscle proteolysis can be blocked by curcumin and that this effect may, at least in part, be caused by inhibited NF-kappaB and p38 activities. The results also suggest that there is not an absolute correlation between changes in muscle protein breakdown rates and changes in atrogin-1 and MuRF1 expression during treatment of muscle wasting. PMID- 18389077 TI - Evolution of a TRIM5-CypA splice isoform in old world monkeys. AB - The TRIM family proteins share a conserved arrangement of three adjacent domains, an N-terminal RING domain, followed by one or two B-boxes and a coiled-coil, which constitutes the tripartite-motif for which the family is named. However, the C-termini of TRIM proteins vary, and include at least nine evolutionarily distinct, unrelated protein domains. Antiviral restriction factor TRIM5alpha has a C-terminal B30.2/SPRY domain, which is the major determinant of viral target specificity. Here, we describe the evolution of a cyclophilin-A encoding exon downstream of the TRIM5 locus of Asian macaques. Alternative splicing gives rise to chimeric transcripts encoding the TRIM motif fused to a C-terminal CypA domain (TRIM5-CypA). We detected TRIM5-CypA chimeric transcripts in primary lymphocytes from two macaque species. These were derived in part from a CypA pseudogene in the TRIM5 locus, which is distinct from the previously described CypA insertion in TRIM5 of owl monkeys. The CypA insertion is linked to a mutation in the 3' splice site upstream of exon 7, which may prevent or reduce expression of the alpha-isoform. All pig-tailed macaques (M. nemestrina) screened were homozygous for the CypA insertion. In contrast, the CypA-containing allele was present in 17% (17/101) of rhesus macaques (M. mulatta). The block to HIV-1 infection in lymphocytes from animals bearing the TRIM5-CypA allele was weaker than that in cells from wild type animals. HIV-1 infectivity remained significantly lower than SIV infectivity, but was not rescued by treatment with cyclosporine A. Thus, unlike owl monkey TRIMCyp, expression of the macaque TRIM5-CypA isoform does not result in increased restriction of HIV-1. Despite its distinct evolutionary origin, Macaca TRIM5-CypA has a similar domain arrangement and shares approximately 80% amino-acid identity with the TRIMCyp protein of owl monkeys. The independent appearance of TRIM5-CypA chimeras in two primate lineages constitutes a remarkable example of convergent evolution. Based on the presence of the CypA insertion in separate macaque lineages, and its absence from sooty mangabeys, we estimate that the Macaca TRIM5-CypA variant appeared 5-10 million years ago in a common ancestor of the Asian macaques. Whether the formation of novel genes through alternative splicing has played a wider role in the evolution of the TRIM family remains to be investigated. PMID- 18389078 TI - Interleukin-6 is crucial for recall of influenza-specific memory CD4 T cells. AB - Currently, our understanding of mechanisms underlying cell-mediated immunity and particularly of mechanisms that promote robust T cell memory to respiratory viruses is incomplete. Interleukin (IL)-6 has recently re-emerged as an important regulator of T cell proliferation and survival. Since IL-6 is abundant following infection with influenza virus, we analyzed virus-specific T cell activity in both wild type and IL-6 deficient mice. Studies outlined herein highlight a novel role for IL-6 in the development of T cell memory to influenza virus. Specifically, we find that CD4+ but not CD8+ T cell memory is critically dependent upon IL-6. This effect of IL-6 includes its ability to suppress CD4+CD25+ regulatory T cells (Treg). We demonstrate that influenza-induced IL-6 limits the activity of virus-specific Tregs, thereby facilitating the activity of virus-specific memory CD4+ T cells. These experiments reveal a critical role for IL-6 in ensuring, within the timeframe of an acute infection with a cytopathic virus, that antigen-specific Tregs have no opportunity to down-modulate the immune response, thereby favoring pathogen clearance and survival of the host. PMID- 18389079 TI - The interferon response inhibits HIV particle production by induction of TRIM22. AB - Treatment of human cells with Type 1 interferons restricts HIV replication. Here we report that the tripartite motif protein TRIM22 is a key mediator. We used transcriptional profiling to identify cellular genes that were induced by interferon treatment and identified TRIM22 as one of the most strongly up regulated genes. We confirmed, as in previous studies, that TRIM22 over expression inhibited HIV replication. To assess the role of TRIM22 expressed under natural inducing conditions, we compared the effects of interferon in cells depleted for TRIM22 using RNAi and found that HIV particle release was significantly increased in the knockdown, implying that TRIM22 acts as a natural antiviral effector. Further studies showed that TRIM22 inhibited budding of virus like particles containing Gag only, indicating that Gag was the target of TRIM22. TRIM22 did not block the release of MLV or EIAV Gag particles. Inhibition was associated with diffuse cytoplasmic staining of HIV Gag rather than accumulation at the plasma membrane, suggesting TRIM22 disrupts proper trafficking. Mutational analyses of TRIM22 showed that the catalytic amino acids Cys15 and Cys18 of the RING domain are required for TRIM22 antiviral activity. These data disclose a pathway by which Type 1 interferons obstruct HIV replication. PMID- 18389080 TI - Adenylyl cyclase alpha and cAMP signaling mediate Plasmodium sporozoite apical regulated exocytosis and hepatocyte infection. AB - Malaria starts with the infection of the liver of the host by Plasmodium sporozoites, the parasite form transmitted by infected mosquitoes. Sporozoites migrate through several hepatocytes by breaching their plasma membranes before finally infecting one with the formation of an internalization vacuole. Migration through host cells induces apical regulated exocytosis in sporozoites. Here we show that apical regulated exocytosis is induced by increases in cAMP in sporozoites of rodent (P. yoelii and P. berghei) and human (P. falciparum) Plasmodium species. We have generated P. berghei parasites deficient in adenylyl cyclase alpha (ACalpha), a gene containing regions with high homology to adenylyl cyclases. PbACalpha-deficient sporozoites do not exocytose in response to migration through host cells and present more than 50% impaired hepatocyte infectivity in vivo. These effects are specific to ACalpha, as re-introduction of ACalpha in deficient parasites resulted in complete recovery of exocytosis and infection. Our findings indicate that ACalpha and increases in cAMP levels are required for sporozoite apical regulated exocytosis, which is involved in sporozoite infection of hepatocytes. PMID- 18389081 TI - An RND-type efflux system in Borrelia burgdorferi is involved in virulence and resistance to antimicrobial compounds. AB - Borrelia burgdorferi is remarkable for its ability to thrive in widely different environments due to its ability to infect various organisms. In comparison to enteric Gram-negative bacteria, these spirochetes have only a few transmembrane proteins some of which are thought to play a role in solute and nutrient uptake and excretion of toxic substances. Here, we have identified an outer membrane protein, BesC, which is part of a putative export system comprising the components BesA, BesB and BesC. We show that BesC, a TolC homolog, forms channels in planar lipid bilayers and is involved in antibiotic resistance. A besC knockout was unable to establish infection in mice, signifying the importance of this outer membrane channel in the mammalian host. The biophysical properties of BesC could be explained by a model based on the channel-tunnel structure. We have also generated a structural model of the efflux apparatus showing the putative spatial orientation of BesC with respect to the AcrAB homologs BesAB. We believe that our findings will be helpful in unraveling the pathogenic mechanisms of borreliae as well as in developing novel therapeutic agents aiming to block the function of this secretion apparatus. PMID- 18389084 TI - Beyond PrP9res) type 1/type 2 dichotomy in Creutzfeldt-Jakob disease. AB - Sporadic Creutzfeldt-Jakob disease (sCJD) cases are currently subclassified according to the methionine/valine polymorphism at codon 129 of the PRNP gene and the proteinase K (PK) digested abnormal prion protein (PrP(res)) identified on Western blotting (type 1 or type 2). These biochemically distinct PrP(res) types have been considered to represent potential distinct prion strains. However, since cases of CJD show co-occurrence of type 1 and type 2 PrP(res) in the brain, the basis of this classification system and its relationship to agent strain are under discussion. Different brain areas from 41 sCJD and 12 iatrogenic CJD (iCJD) cases were investigated, using Western blotting for PrP(res) and two other biochemical assays reflecting the behaviour of the disease-associated form of the prion protein (PrP(Sc)) under variable PK digestion conditions. In 30% of cases, both type 1 and type 2 PrP(res) were identified. Despite this, the other two biochemical assays found that PrP(Sc) from an individual patient demonstrated uniform biochemical properties. Moreover, in sCJD, four distinct biochemical PrP(Sc) subgroups were identified that correlated with the current sCJD clinico pathological classification. In iCJD, four similar biochemical clusters were observed, but these did not correlate to any particular PRNP 129 polymorphism or western blot PrP(res) pattern. The identification of four different PrP(Sc) biochemical subgroups in sCJD and iCJD, irrespective of the PRNP polymorphism at codon 129 and the PrP(res) isoform provides an alternative biochemical definition of PrP(Sc) diversity and new insight in the perception of Human TSE agents variability. PMID- 18389082 TI - Hepatocyte permissiveness to Plasmodium infection is conveyed by a short and structurally conserved region of the CD81 large extracellular domain. AB - Invasion of hepatocytes by Plasmodium sporozoites is a prerequisite for establishment of a malaria infection, and thus represents an attractive target for anti-malarial interventions. Still, the molecular mechanisms underlying sporozoite invasion are largely unknown. We have previously reported that the tetraspanin CD81, a known receptor for the hepatitis C virus (HCV), is required on hepatocytes for infection by sporozoites of several Plasmodium species. Here we have characterized CD81 molecular determinants required for infection of hepatocytic cells by P. yoelii sporozoites. Using CD9/CD81 chimeras, we have identified in CD81 a 21 amino acid stretch located in a domain structurally conserved in the large extracellular loop of tetraspanins, which is sufficient in an otherwise CD9 background to confer susceptibility to P. yoelii infection. By site-directed mutagenesis, we have demonstrated the key role of a solvent-exposed region around residue D137 within this domain. A mAb that requires this region for optimal binding did not block infection, in contrast to other CD81 mAbs. This study has uncovered a new functionally important region of CD81, independent of HCV E2 envelope protein binding domain, and further suggests that CD81 may not interact directly with a parasite ligand during Plasmodium infection, but instead may regulate the function of a yet unknown partner protein. PMID- 18389085 TI - Drug- and region-specific effects of protracted antidepressant and cocaine treatment on the content of melatonin MT(1) and MT(2) receptor mRNA in the mouse brain. AB - OBJECTIVES: In the mammalian brain, G protein-coupled MT(1) and MT(2) melatonin receptors may be involved in Alzheimer's pathology, long-term potentiation, depression, and in the behavioral effects of psychoactive drugs. These drugs; e.g. antidepressants and drugs of abuse, are typically used over long periods of time and may alter neuroplasticity and gene expression. We hypothesized that such antidepressant- and cocaine-altered expression of melatonin receptor mRNA may occur in the hippocampus and striatum. METHODOLOGY: Male C3H/HeJ mice were treated with the antidepressants fluoxetine, desipramine, and clomipramine, with the psychostimulant cocaine, and with a vehicle either a single time or once a day for 14 days. Brain samples were collected 24 h after the last injection and the content of MT(1) and MT(2) mRNA was assayed. RESULTS: A single drug injection did not alter the MT(1) and MT(2) mRNA content. In the hippocampus, protracted treatment with antidepressants increased the amount of MT(1) mRNA (with the exception of fluoxetine) but decreased MT(2) mRNA content; cocaine did not produce any alterations. In the striatum, antidepressants produced the opposite effect on MT(1) mRNA content; they decreased it. They did not significantly alter striatal MT(2) mRNA (we observed a nonsignificant trend to a decrease). Cocaine also decreased striatal MT(1) mRNA content without affecting MT(2) mRNA. CONCLUSION: These results suggest that drug- and region-specific alterations of MT(1)/MT(2) mRNA produced by protracted antidepressants and cocaine treatment may alter MT1/MT2 expression and contribute to long-term neuroplastic effects of these drugs. PMID- 18389086 TI - Articulatory characteristics of Hungarian 'transparent' vowels. AB - Using a combination of magnetometry and ultrasound, we examined the articulatory characteristics of the so-called 'transparent' vowels [i], [i], and [e] in Hungarian vowel harmony. Phonologically, transparent vowels are front, but they can be followed by either front or back suffixes. However, a finer look reveals an underlying phonetic coherence in two respects. First, transparent vowels in back harmony contexts show a less advanced (more retracted) tongue body posture than phonemically identical vowels in front harmony contexts: e.g. [i] in buli val is less advanced than [i] in bili-vel. Second, transparent vowels in monosyllabic stems selecting back suffixes are also less advanced than phonemically identical vowels in stems selecting front suffixes: e.g. [i] in ir, taking back suffixes, compared to [i] of hir, taking front suffixes, is less advanced when these stems are produced in bare form (no suffixes). We thus argue that the phonetic degree of tongue body horizontal position correlates with the phonological alternation in suffixes. A hypothesis that emerges from this work is that a plausible phonetic basis for transparency can be found in quantal characteristics of the relation between articulation and acoustics of transparent vowels. More broadly, the proposal is that the phonology of transparent vowels is better understood when their phonological patterning is studied together with their articulatory and acoustic characteristics. PMID- 18389087 TI - Rarity of Somatic Mutation and Frequency of Normal Sequence Variation Detected in Sporadic Colon Adenocarcinoma Using High-Throughput cDNA Sequencing. AB - We performed high-throughput cDNA sequencing in colorectal adenocarcinoma and matching normal colorectal epithelium. All six hundred three genes in the UCSC database that were expressed in colon cancers and contained open reading frames of 1000 nucleotides or less were selected for study (total basepairs/bp, 366,686). 304,350 of these 366,686 bp (83.0%) were amplified and sequenced successfully. Seventy-eight sequence variants present in germline (i.e. normal) as well as matching somatic (i.e. tumor) DNA were discovered, yielding a frequency of 1 variant per 3,902 bp. Fifty-one of these sequence variants were homozygous (26 synonymous, 25 non-synonymous), while 27 were heterozygous (11 synonymous, 16 non-synonymous). Cancer tissue contained only one sequence-altered allele of the gene ATP50, which was present heterozygously alongside the wild type allele in matching normal epithelium. Despite this relatively large number of bp and genes sequenced, no somatic mutations unique to tumor were found. High throughput cDNA sequencing is a practical approach for detecting novel sequence variations and alterations in human tumors, such as those of the colon. PMID- 18389088 TI - Retapamulin: a new topical antibiotic for the treatment of uncomplicated skin infections. AB - Retapamulin is the first agent in the new pleuromutilin class of antibacterials to become commercially available for clinical use in humans. Retapamulin acts as a potent inhibitor of bacterial protein synthesis and has a unique mode of antibiotic action. To date, retapamulin has not demonstrated any clinically relevant, target-specific crossresistance with other antibiotic classes, and has shown a low potential for resistance selection in vitro. In preclinical studies, retapamulin demonstrated pronounced in vitro activity against staphylococcal, streptococcal and anaerobic Gram-positive clinical isolates associated with skin and skin structure infections. Clinical pharmacology studies showed low systemic exposure with topical use of retapamulin, and a favorable tolerability profile. In clinical efficacy trials involving pediatric and adult patients who received retapamulin twice daily for five days, retapamulin was highly effective in the treatment of impetigo, secondarily infected traumatic lesions and secondarily infected dermatitis. Further, the clinical efficacy and safety profile of retapamulin was comparable to that of commonly used oral and topical antibiotics. Retapamulin was also clinically effective against isolates resistant to existing therapies. As a 1% ointment, retapamulin has been approved in the United States for the treatment of impetigo and in Europe for the shortterm treatment of impetigo and infected small lacerations, abrasions and sutured wounds. PMID- 18389089 TI - Atazanavir/ritonavir: a review of its use in HIV therapy. AB - Recommendations for a highly active antiretroviral therapy (HAART) in either pretreated patients or symptomatic patients with an AIDS-defining event are based on a combination of three or more agents from different antiretroviral classes including two nucleoside reverse transcriptase inhibitors with at least one protease inhibitor. The majority of currently available protease inhibitors are coadministered with low-dose ritonavir as a pharmacoenhancer that significantly increases protease inhibitor plasma concentrations. Atazanavir is a highly active azapeptide inhibitor of the HIV protease. It was the first, and to date the only, protease inhibitor designed to be applied once daily (q.d.) and is expected to overcome the problems of earlier agents of this class of drugs, such as unfavorable adverse events like hyperlipidemia, diarrhea and lipodystrophy. Atazanavir, formerly known as BMS-232632, can be dosed either at 400 mg q.d. without a pharmacoenhancer as first-line HIV therapy or combined with ritonavir as atazanavir/ritonavir 300/100 mg q.d. for therapy-experienced patients. The pharmacoenhancing effect of ritonavir on atazanavir resulted in a potent, clinically effective and well-tolerated antiretroviral drug with high plasma concentrations and a sufficient genetic barrier to viral resistance. Nevertheless, noninferiority to lopinavir/ritonavir-containing HAART could not be shown when atazanavir was given unboosted in pretreated patients in the AI424-043 study. When atazanavir was boosted with low-dose ritonavir its efficacy was comparable to that of lopinavir/ritonavir in non-naive patients (AI424-045 study). Additionally, specific side effects were identified during clinical practice, such as an increased rate of patients with jaundice, and, more recently, genetic risk factors causing hyperbilirubinemia. Atazanavir inhibits glucuronyltransferase, an enzyme responsible for the metabolism of bilirubin in liver, thus increasing unconjugated bilirubin levels in blood. However, atazanavir itself also enhances plasma concentrations of other coadministered HIV 1 protease inhibitors, so that its use as a combination partner in boosted double protease inhibitor combinations, with or without the addition of nucleoside reverse transcriptase inhibitors, is being evaluated. Unboosted atazanavir is approved for first-line HIV therapy in adults in the United States, and atazanavir/ritonavir is recommended for the second-line therapy of HIV-1 infection in adult HIV-1-infected patients in the United States and the European Union. More recently, data from the CASTLE study (AI424-138) have been reported at the 15th Conference on Retroviruses and Opportunistic Infections by Molina et al., where boosted atazanavir-containing HAART was compared to a regimen with lopinavir/ritonavir in therapy-naive patients. PMID- 18389091 TI - An evidence-based review updating the various treatment and management approaches to serotonin reuptake inhibitor-associated sexual dysfunction. AB - Although most patients (80-90%) will respond to the first or second antidepressant prescribed, major depression disease management outcomes data are poor. Serotonin reuptake inhibitor (SRI) antidepressant-associated sexual dysfunction, which is reported to occur in 40-70% of patients on these agents, is a major factor for treatment noncompliance, treatment failure and costly disease management outcomes. Up to 90% of patients with treatment-emergent sexual dysfunction will discontinue their prescribed medication prematurely. Despite several thousand published reports on treatment modalities based on heuristic post hoc hypotheses of central serotonin inhibition and those involving agonist, antagonist, partial agonist, switching, augmentation and waiting management approaches, no evidence-based data are available to support those treatment modalities, leaving patients exposed to random pharmacology. The emergence of new approaches based on novel signaling to treat sexual dysfunction, which demonstrate efficacy for selective type 5 phosphodiesterase inhibitor treatment of SRI antidepressant-associated sexual dysfunction, offers an opportunity for an evidence-based re-evaluation of the comparative effectiveness of various management approaches to SRI antidepressant-associated sexual dysfunction. PMID- 18389090 TI - Drospirenone/ethinyl estradiol. AB - Drospirenone 3 mg/ethinyl estradiol 20 microg (24/4) is a new unique oral contraceptive formulation that combines in a novel dosing regimen the lowest dosage of ethinyl estradiol commonly used today with drospirenone, an innovative progestin. Drospirenone is a compound closely resembling progesterone, but with the antimineralocorticoid and antiandrogenic properties of a related therapeutic agent, the diuretic, antihypertensive and androgen receptor antagonist, 17alpha spironolactone. The prolongation of hormonally active pills in the monthly drospirenone/ethinyl estradiol cycle from 21 days to 24 days, followed by 4 days of inactive pills, is an interesting variant of the recently developed extended pill regimens (1). Recent contraceptive research has focused on improving side effect profiles and providing noncontraceptive health and lifestyle advantages. Many of these benefits are now supported with evidence-based medicine (2). Most available oral contraceptives improve cycle regularity, menstrual pain, excessive menstrual flow and acne. However, weight gain, bloating, food cravings, breast tenderness and mood alterations (especially irritability and depression and the complex of affective, behavioral and somatic symptoms of premenstrual syndrome [PMS] and the severe form of PMS, premenstrual dysphoric disorder [PMDD]) are not generally improved with the traditional oral contraceptive formulations (3). Drospirenone/ethinyl estradiol 24/4 is currently the only hormonally based contraceptive regimen with large, randomized, controlled trials demonstrating efficacy for PMDD. It has received U.S. Food and Drug Administration (FDA) indications not only for the prevention of pregnancy but also for PMDD and for moderate acne vulgaris in women who choose oral contraception for birth control (4, 5). PMID- 18389092 TI - Gender-dependent differences in the response of human adipocytes to leptin lipolytic action. AB - Lipid turnover and deposition is under the control of nutritional, developmental, metabolic and neuroendocrine influences. The aim of this investigation was to study whether there is a gender-dependent difference in the effect of leptin in lipolytic activity of freshly isolated human fat cells. The lipolytic rate was assessed through the glycerol release after incubation with leptin at different concentrations ranging from 10(-6) to 10(-12) M in isolated adipocytes obtained from male and female omental adipose tissue. The relative ED(50) concentration of leptin was found to be 4.42 x 10(-10) M in adipocytes from men and 3.08 x 10(-12) M in fat cells from women. The presence of leptin at concentrations of 10(-12) to 10(-6) M in the incubation medium of isolated fat cells significantly increased glycerol release when compared with basal levels, being always higher (8-15%) in female adipocytes when compared with male specimens for all tested concentrations when expressed on a fat content relative basis. This study demonstrates gender differences regarding the lipolytic actions of leptin, which may help the pharmacological application of leptin-based therapies. PMID- 18389093 TI - Radioprotective effects of chlorogenic acid against mortality induced by gamma irradiation in mice. AB - The radioprotective effects of the naturally occurring compound chlorogenic acid have been investigated against mortality induced by gamma-irradiation in mice. Chlorogenic acid was administrated at single doses of 100, 200 and 400 mg/kg 1 or 24 h prior to lethal dose of gamma-irradiation (8.5 Gy). At 30 days after treatment, the percentage of survival in each group was as follows: control, 20%; 100 mg/kg, 20% and 15%; 200 mg/kg, 45% and 15%; 400 mg/kg, 25% and 35% for 1 h and 24 h treatment prior gamma-irradiation, respectively. Survival rate was statistically increased in animals treated with this agent at 200 mg/kg at 1 h prior to irradiation as compared with the irradiation only group. Other doses of chlorogenic acid have not showed any enhanced survival when it was administrated at 1 or 24 h before irradiation. Chlorogenic acid exhibited concentration dependent activity on 1,1-diphenyl-2-picrylhydrazyl free radical to show strong antioxidant activity. It appeared that chlorogenic acid with antioxidant activity reduced mortality induced by gamma-irradiation. PMID- 18389094 TI - Initiation of apoptosis by photodynamic therapy using a novel positively charged and water-soluble near infra-red photosensitizer and white light irradiation. AB - Our aim was to investigate the photophysical and photodynamic properties of a new, water-soluble positively charged and chemically stable photosensitizer: tetrahydroporphyrin tetratosylat (THPTS). Absorption, fluorescence and (1)H NMR spectra and the intracellular distribution of THPTS were measured. The apoptosis in choroidal melanoma cells was measured using cell death detection ELISA and caspase-8 activity assay. THPTS-PDT efficiency was studied in Balb/c mice bearing C26 colon carcinoma. Subcutaneously located tumors were irradiated with a white light source at a fluence rate of 100 mW/cm(2). THPTS was administrated 3 h before illumination. The tumoricidal effect was examined 24 h after THPTS-PDT by vital staining with 0.4-ml 1% Evans blue solution, intraperitoneally injected to each mouse. THPTS showed a strong absorption band at 760 nm. Its purity, measured by (1)H NMR, is better than 99%. At 24-h incubation period, CLSM revealed THPTS fluorescence in mitochondria and cell nucleus. THPTS possesses no toxic effect in preincubated CM cells without irradiation, and THPTS-PDT causes efficient apoptosis. THPTS-PDT using white light irradiation at a dose of 480 J/cm(2) caused necrosis with a depth of 8 mm in subcutaneously located C26 colon carcinoma in Balb/c-mice. In accordance with the present results, the THPTS seems to be of interest for further in vivo investigations with broad-band white light sources. PMID- 18389095 TI - Effects of Persea americana Mill (Lauraceae) ["Avocado"] ethanolic leaf extract on blood glucose and kidney function in streptozotocin-induced diabetic rats and on kidney cell lines of the proximal (LLCPK1) and distal tubules (MDBK). AB - Extracts of Persea americana Mill (Lauraceae) ("Avocado") have been traditionally used to treat hypertension and diabetes mellitus. Accordingly, we studied the hypoglycaemic and renal function effects of P. americana leaf ethanolic extracts (PAE) in STZ-induced diabetic rats. Oral glucose tolerance responses to various doses of PAE were monitored in fasted rats following a glucose load. Rats treated with deionized water or standard hypoglycaemic drugs acted as untreated and treated positive controls, respectively. Acute renal effects of PAE were investigated in anesthetized rats challenged with 0.077 M NaCl after a 3.5-h equilibration for 4 h comprising 1 h control, 1.5 h treatment and 1.5 h recovery periods. PAE was added to the infusate during the treatment period. Hepatic glycogen concentration was measured after 6 weeks of daily treatment with PAE. PAE induced dose-dependent hypoglycaemic responses in STZ-induced diabetic rats while subchronic PAE treatment additionally increased hepatic glycogen concentrations. Acute PAE infusion decreased urine flow and electrolyte excretion rates, whilst subchronic treatment reduced plasma creatinine and urea concentrations. These results indicate not only the basis of the ethnomedicinal use of P. americana leaf extract in diabetes management, but also of need for further studies to identify and evaluate the safety of PAE's bioactive compounds. PMID- 18389096 TI - Study of paracetamol 1-g oral solution bioavailability. AB - The aim of this work was to assess paracetamol bioavailability after administering 1 g in oral solution. Eighteen healthy volunteers were selected for this open-label study. A total of 15.4 ml of Gelocatil Oral Solution (Laboratorios Gelos, S.L.), corresponding to 1 g of paracetamol, were administered to fasting subjects. Blood samples were collected at 0 min, 10 min, 20 min, 30 min, 45 min, 1 h, 1.5 h, 2 h, 3 h, 4 h, 6 h, 8 h, 10 h and 12 h. Paracetamol plasma concentrations were determined by reverse-phase high performance liquid chromatography. The study was conducted without deviations from protocol. Pharmacokinetic data from 18 subjects were allowed for estimating fast and high-paracetamol bioavailability: t(max) 20 min (10-45) [median (range)], C(max) 24. 3 mg/l (6.5) [mean (standard deviation)], AUC(0-t) 64.0 mg h/l (16.1) and AUC(0-00) 68.1 mg h/l (17.9). These results are comparable to those described for Gelocatil Oral Solution given at a 650 mg dose and for immediate release Gelocatil 650 mg tablets. Absorption speed was very fast, similar to that described for other oral-solution formulations, which provides an immediate onset of pain and fever relief. The results of this study show suitable bioavailability for 1 g Gelocatil Oral Solution, with fast-absorption speed that provides an immediate onset of pain and fever relief. PMID- 18389097 TI - Apoptotic mechanisms involved in neurodegenerative diseases: experimental and therapeutic approaches. AB - Alzheimer's disease (AD) and Parkinson's disease (PD) are two of the most significant neurodegenerative disorders in the developed world. However, although these diseases were described almost a century ago, the molecular mechanisms that lead to the neuronal cell death associated with these diseases are not yet clear, and vigorous research efforts have failed to identify effective treatment options. In the present review, we evaluate the potential mechanisms underlying apoptosis and neuronal death in neurodegenerative disorders. A role for mitochondria in the release of proapoptotic proteins, such as cytochrome c and apoptosis-inducing factor (AIF) etc., is discussed along with key processes involving oxidative stress and activation of glutamate receptors. We also deliberate the implication of DNA damage, primarily p53 induction and reentry in the cell cycle. Finally, we postulate that multitargeting therapies comprising antioxidants, cell cycle inhibitors and modulating agents of COX-2 or c-JUN kinase pathways could be suitable strategies to prevent or delay the process of neuronal cell death in neurodegenerative disorders. Thus, the aim of this review is to discuss the pathways involved in the pathogenesis of neurodegenerative diseases such as AD, PD and Huntington's disease (HD). Furthermore, current and future pharmacotherapeutics will be considered. PMID- 18389098 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials are a guide to the most recent clinical trials in current literature and congresses. The data in the following tables has been retrieved from the Clinical Trials Knowledge Area of Prouse Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: 101M, 3F8; Abatacept, ABT-263, Adalimumab, AG-7352, Agatolimod sodium, Alfimeprase, Aliskiren fumarate, Alvimopan hydrate, Aminolevulinic acid hexyl ester, Ammonium tetrathiomolybdate, Anakinra, Aripiprazole, AS-1404, AT-9283, Atomoxetine hydrochloride, AVE-1642, AVE-9633, Axitinib, AZD-0530; Becocalcidiol, Belotecan hydrochloride, Bevacizumab, BG-9928, BIBF-1120, BMS-275183, Bortezomib, Bosentan; Catumaxomab, Cetuximab, CHR-2797, Ciclesonide, Clevidipine, Cypher, Cytarabine/daunorubicin; Darifenacin hydrobromide, Darunavir, Denosumab, Desvenlafaxine succinate, Disufenton sodium, Duloxetine hydrochloride, Dutasteride; Eculizumab, Efalizumab, Eicosapentaenoic acid/docosahexaenoic acid, Eplerenone, Epratuzumab, Erlotinib hydrochloride, Escitalopram oxalate, Ethynylcytidine, Etravirine, Everolimus, Ezetimibe; Fulvestrant; Garenoxacin mesilate, Gefitinib, Gestodene; HI-164, Hydralazine hydrochloride/isosorbide dinitrate; Icatibant acetate, ICX-RHY, Idraparinux sodium, Indacaterol, Ispronicline, Ivabradine hydrochloride, Ixabepilone; KB-2115, KW-2449; L-791515, Lapatinib ditosylate, LGD-4665, Licofelone, Liposomal doxorubicin, Lisdexamfetamine mesilate, Lumiracoxib; Methoxy polyethylene glycol-epoetin-beta, Miglustat, Mipomersen sodium, Mitumprotimut-T, MK-0822A, MK-0974; Nelarabine; Obatoclax mesylate, Olmesartan medoxomil, Olmesartan medoxomil/hydrochlorothiazide; Paliperidone, Palonosetron hydrochloride, Panitumumab, Pegfilgrastim, Peginterferon alfa-2a, Pemetrexed disodium, Perospirone hydrochloride, Pertuzumab, Pimecrolimus, Pitrakinra, Pixantrone maleate, Posaconazole, Pregabalin; Quercetin; RALGA, Raltegravir potassium, Ranelic acid distrontium salt, rhHistone 1.3, Rimonabant, Rivaroxaban, Rosuvastatin calcium, RTS,S/SBAS2; Satraplatin, SNDX-275, Sodium butyrate, Solifenacin succinate, Sorafenib, SU-14813, Sunitinib malate; Tadalafil, Tafenoquine succinate, Tamatinib fosdium, Taxus, Telbivudine, Telmisartan/hydrochlorothiazide, Temsirolimus, Tiotropium bromide, Tipranavir, Tocilizumab, Trabectedin, Tramadol hydrochloride/acetaminophen; Ulipristal acetate, Uracil, Ursodeoxycholyltaurine; Valdecoxib, Vardenafil hydrochloride hydrate, Varenicline tartrate, Vildagliptin, Vinflunine, Vitespen, Vorinostat; ZK EPO, Zoledronic acid monohydrate. PMID- 18389099 TI - The antiviral activity of Toll-like receptor 7 and 7/8 agonists. AB - Imiquimod 5% cream is approved for the topical treatment of external anogenital warts caused by human papillomavirus (HPV) and for the skin cancer conditions superficial basal cell carcinoma and actinic keratosis. This drug is the first approved topically active Toll-like receptor (TLR) 7 agonist. Imiquimod activates innate immune cells to produce interferon-a and other cytokines. The induced cytokine cascade, in combination with effects in enhancing antigen presentation, also promotes an antigen-specific T helper type 1 cell-mediated immune response. This immune-based mechanism provides activity against a number of viruses and other intracellular pathogens. Imiquimod was effective topically in clinical studies for HPV but caused mixed results for Molluscum contagiosum, and herpes simplex virus (HSV). Activity against several other viruses were reported in case reports or patient series involving "off-label" usage of imiquimod, while others were evaluated only in preclinical models. Resiquimod, a more potent investigational analogue of imiquimod with mixed TLR7/8 agonist activity, was evaluated in clinical studies topically for the treatment of HSV and systemically for hepatitis C virus also with mixed success. This review focuses on the mechanism of action and antiviral usage reported for the TLR7 agonist imiquimod, the TLR7/8 agonist resiquimod and related imidazoquinoline analogues. PMID- 18389101 TI - Angiogenesis: the new potential target for the therapy of psoriasis? AB - Angiogenesis is a hallmark of chronic inflammation such as psoriasis. Unraveling the pathogenesis of psoriasis shows that several proangiogenic mediators are activated and highly expressed during psoriasis. Vascular endothelial growth factor, hypoxia- inducible factor, tumor necrosis factor, interleukin-8 and angiopoietins are considered to be the main players responsible for the strong vessel formation in psoriasis. The proangiogenic milieu in the skin seems to result from a proinflammatory immune response initiated by T helper cells. Interestingly, several small molecules as well as modern biologics used for systemic therapy of psoriasis have been shown to provide not only immune regulatory effects but also influence endothelial cell biology. Thus, direct targeting of angiogenesis may not only help to understand psoriasis pathogenesis but also to develop new strategies to treat psoriasis with therapeutics that halt the angiogenesis required for the inflammatory disease. PMID- 18389102 TI - The urokinase plasminogen activator system in cancer: a putative therapeutic target? AB - The urokinase plasminogen activator (uPA) system is an endogenous proteolytic cascade which can be actively subverted by the neoplastic process to facilitate progression and metastasis. Abundant experimental and clinical evidence supports such a role and elevated levels of components of the uPA system are strong negative prognosis indicators in a wide variety of tumor types. Collectively this data makes the uPA system an attractive option for targeted intervention. This review examines the role of the uPA system in tumor invasion and metastasis and assess the various therapeutic modalities developed to selectively exploit this system. PMID- 18389100 TI - The role of sphingosine-1-phosphate and its receptors in asthma. AB - Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite that plays important roles in allergic responses, including asthma and anaphylaxis, the incidence of which is rising worldwide especially in industrialized urban populations. In this review, we will discuss how S1P is formed and released, and how it acts at many cellular levels, including mast cells, the airway epithelium, airway smooth muscle and many immune cells. Since the actions of S1P on all of these cells could exacerbate allergic responses, the proteins that synthesize, release and respond to S1P offer plausible targets for a new generation of antiinflammatory therapeutics. PMID- 18389103 TI - Molecule of the month. Elesclomol and obatoclax mesylate. PMID- 18389104 TI - Advances in antiretroviral therapy. AB - The 4th International AIDS Society (IAS) Conference on HIV Pathogenesis, Treatment and Prevention (IAS 2007), was held in Sydney, Australia, from July 22 25, 2007. The data presented at IAS 2007 were categorized into three tracks: Basic science; clinical research, treatment and care; and biomedical prevention. This article focuses on presentations at the conference that directly dealt with antiretroviral therapy. PMID- 18389105 TI - Polymer assisted deposition. AB - Polymer assisted deposition (PAD) is a chemical solution route to high quality thin films of metal oxides. This technique employs metal ions coordinated to polymers as the film precursor. The use of polymer bound metals has several advantages. The polymer controls the viscosity and binds metal ions, resulting in a homogeneous distribution of metal precursors in the solution and the formation of uniform metal oxide films. The nature of the metal oxide deposition is dominated by bottom-up growth, leading to ready formation of crack-free epitaxial metal oxides and the ability to coat nanofeatured substrates in a conformal fashion. PMID- 18389106 TI - A three-dimensional metal-organic framework with a distorted Kagome related layer showing canted antiferromagnetic behaviour. AB - Three-dimensional [Mn(3){C(6)H(3)(COO)(3)}(2)] comprising two-dimensional -Mn-O Mn- distorted Kagome layers, formed by two geometrically different hexa coordinated Mn(2+) ions, show less frustration and spin canted long range ordering possibly due to antisymmetric exchange. PMID- 18389107 TI - Anion induced and inhibited circumrotation of a [2]catenane. AB - The first example of a catenane capable of performing circumrotation via an anion switching methodology is described; of particular interest is a conformational locking mechanism which results from chloride coordination in the catenane binding cavity. PMID- 18389108 TI - Fabrication of optically active flexible polymer films with embedded chain-like arrays of silver nanoparticles. AB - We report a straightforward method for preparation of freestanding transparent polymer film containing 2D silver nanoparticle arrays and possessing polarization sensitive optical properties. PMID- 18389109 TI - Generation of a solid Bronsted acid site in a chiral framework. AB - Protonation of chiral porous materials introduces a Bronsted acid centre, the structure of which is unique to the heterogeneous phase requiring pore wall confinement for stable isolation. PMID- 18389110 TI - Direct electrochemistry and electrocatalytic mechanism of evolved Escherichia coli cells in microbial fuel cells. AB - E. coli cells evolved under electrochemical tension in a microbial fuel cell possess direct electrochemical behavior due to the excretion of hydroquinone derivatives through a highly permeable outer membrane, and their catalyzed fuel cell demonstrates excellent performance. PMID- 18389112 TI - Controllable preparation of Zn(II) coordination polymers: unusual solvothermal formation of a LiGe-type framework directed by in situ S-S coupling of 5-(4 pyridyl)-1H-1,2,4-triazole-3-thiol. AB - Controlled assembly of Zn(II) with a versatile ligand 5-(4-pyridyl)-1H-1,2,4 triazole-3-thiol (Hptt) in conventional or solvothermal condition affords two distinct coordination frameworks, in which the former shows a unique 3-D pseudo polyrotaxane architecture with 1-D water tapes penetrating the 2-D Zn-ptt layers, whereas the later is generated from in situ S-S coupling of Hptt and has a very rare 2-fold interpenetrating LiGe (lig) topology. PMID- 18389111 TI - Highly active and stereoselective zirconium and hafnium alkoxide initiators for solvent-free ring-opening polymerization of rac-lactide. AB - Under solvent-free conditions (at 130 degrees C), zirconium and hafnium amine tris(phenolate) alkoxides are extremely active, well-controlled, single-site initiators for the ring-opening polymerization of rac-lactide, yielding highly heterotactic polylactide. PMID- 18389113 TI - New supramolecular isomers with 2D 4(4) square-grid and 3D 6(5).8 frameworks in a one-pot synthesis; reversible solvent uptake and intriguing luminescence properties. AB - Two supramolecular isomers of [Ni(4-bpd)(2)(NCS)(2)] (4-bpd = 1,4-bis(4-pyridyl) 2,3-diaza-1,3-butadiene) with 2D 4(4) square-grid and 3D 6(5).8 frameworks are co crystallized in a one-pot reaction, both of which exhibit interesting luminescence properties and reversible adsorption-desorption with respect to guest solvents. PMID- 18389114 TI - Conjugated polyelectrolyte-DNA complexes for multi-color and one-tube SNP genotyping assays. AB - The complexes of a cationic conjugated polymer with DNA are designed as new platforms for homogeneous, sensitive and facile fluorescence assays for SNP genotyping, which interface with single-base extension, multi-step FRET and optical amplification properties of conjugated polymers. PMID- 18389115 TI - Electronic structure of the mononuclear Ag(ii) complex [Ag([18]aneS(4)O(2))](2+) ([18]aneS(4)O(2) = 1,10-dioxa-4,7,13,16-tetrathiacyclooctadecane). AB - The structure of [Ag([18]aneS(4)O(2))](PF(6))(2).CH(2)Cl(2) shows a highly unusual and unexpected boat conformation for the macrocycle with square-planar S(4)-coordination at the formal Ag(ii) centre and the two ether O-centres lying on the same side of the S(4) plane; the SOMO in [Ag([18]aneS(4)O(2))](2+) possesses 22.7% Ag 4d(xy) character, as determined by multi-frequency EPR spectroscopy and supported by DFT calculations. PMID- 18389116 TI - Site-specific immobilization of CMP-sialic acid synthetase on magnetic nanoparticles and its use in the synthesis of CMP-sialic acid. AB - Through the native chemical ligation, CMP-sialic acid synthetase (CSS) was site specifically immobilized on magnetic nanoparticles and presented excellent enzymatic performance. PMID- 18389117 TI - An EPR method for measuring the rate of distribution of organic substrates between cyclodextrin, micelles and water. AB - The combined use of selected nitroxides and EPR spectroscopy has been proved to be suitable for studying the partitioning rate of a given substrate in cyclodextrin-micelle systems. PMID- 18389118 TI - Synthesis and characterization of tris(2-pyridylthio)methanido Zn complex with a Zn-C bond and DFT calculation of its one-electron oxidized species. AB - Tris(2-pyridylthio)methane (tptmH) reacts with ZnCl(2) producing the Zn-C containing complex of [ZnCl(tptm)], whose cyclic voltammogram shows an irreversible oxidation peak at 0.2 V vs. E(0')(Fc(+/0)). DFT calculations suggested that 1e(-) oxidation should occur at the tptm ligand resulting in the cleavage of the Zn-C bond, leading to decomposition of the complex. PMID- 18389119 TI - Inkjet fabrication of hydrogel microarrays using in situ nanolitre-scale polymerisation. AB - Polymer hydrogel microarrays were fabricated by inkjet printing of monomers and initiator, allowing up to 1800 individual polymer features to be printed on a single glass slide. PMID- 18389120 TI - Click-functionalization of conducting poly(3,4-ethylenedioxythiophene) (PEDOT). AB - Efficient post-functionalization of conductive polymer films was achieved by Cu(+)-catalyzed "click"-cycloaddition of novel poly(azidomethyl-EDOT) and various functionalized terminal alkynes under mild heterogeneous conditions with high conversion efficiencies. PMID- 18389121 TI - Molecular folding screen: folding and unfolding of 1,8-anthrylene-ethynylene oligomers by photochemical cycloaddition and thermal cycloreversion. AB - Molecular folding screens consisting of anthracene plates and acetylene linkers stereoselectively fold into a zigzag form by [4 + 4]photocycloaddition, and unfold by thermal cycloreversion. PMID- 18389122 TI - pi-Conjugated macrocycles from thiophenes and benzenes. AB - pi-Conjugated macrocycles consisting of thiophenes and benzenes exhibit benzenoid features for 4npi macrocycles, whereas (4n + 2)pi macrocycles are annulenoid due to rapid interconversion between quinoid and Kekule canonical forms in the benzene units. PMID- 18389123 TI - UV-induced immobilization of tethered zirconocenes on H-terminated silicon surfaces. AB - A tethered ethylenebis(indenyl) zirconocene was covalently immobilized on H terminated Si(111) surfaces using UV-mediated alkene hydrosilylation, thus making possible the development of structured catalytic surfaces with highly controlled properties. PMID- 18389124 TI - Cu-catalyzed regioselective carbomagnesiation of dienes and enynes with sec- and tert-alkyl Grignard reagents. AB - The carbomagnesiation of dienes and enynes with sec- and tert-alkyl Grignard reagents has been achieved by using copper salts as catalysts. PMID- 18389125 TI - Diastereoselective diaza-Cope rearrangement reaction. AB - Steric effect is used to obtain a highly diastereoselective rearrangement reaction. PMID- 18389126 TI - STEM characterization on silica nanowires with new mesopore structures by space confined self-assembly within nano-scale channels. AB - "Critical" channel diameters were found (below which space confinement takes effect, leading to more uniform and ordered mesopore structures) in the study of evaporation-induced co-assembly of triblock-copolymer (P123) and silica molecular precursors (TEOS, tetraethyl orthosilicate) by employing channels in anodized aluminum oxide (AAO, 13-200 nm channel diameter) and in track-etched polycarbonate (EPC, 10-80 nm channel diameter) and for the first time we have observed a new mesopore structure (i.e., packed hollow spheres) in silica nanowires formed in AAO channels with diameters from 30 to 80 nm. PMID- 18389127 TI - Versatile and efficient functionalisation of multiallylic dendronised polymers: can dense packing be reached? AB - Surface modification of a multiallylic dendronised polymer was performed using hydrosilylation, hydroboration or radical addition of thiols to cover the polymer with various functional moieties; surface congestion is expected to occur when bulky groups are grafted. PMID- 18389128 TI - Photoisomerization of the green fluorescence protein chromophore and the meta- and para-amino analogues. AB - The Z --> E photoisomerization and fluorescence quantum yields for the wild-type green fluorescence protein (GFP) chromophore (p-HBDI) and its meta- and para amino analogues (m-ABDI and p-ABDI) in aprotic solvents (hexane, THF, and acetonitrile) and protic solvents (methanol and 10-20% H(2)O in THF) are reported. The dramatic decrease in the quantum yields on going from aprotic to protic solvents indicates the important role of solvent-solute hydrogen bonding in the nonradiative decay pathways. The enhanced fluorescence of m-ABDI is also discussed. PMID- 18389129 TI - Nickeladihydrofuran. Key intermediate for nickel-catalyzed reaction of alkyne and aldehyde. AB - The formation of a nickeladihydrofuran by oxidative cyclization of an alkyne and an aldehyde with nickel(0) has been demonstrated; the transformation of the nickeladihydrofuran into an enone by decomposition, a lactone by carbonylation and an allylic alcohol by treatment with ZnMe(2) suggests that nickeladihydrofuran is an important key intermediate in a variety of catalytic reactions. PMID- 18389130 TI - New palladium-catalyzed aerobic oxidative cleavage and cyclization of N-aryl peptide derivatives. AB - Oxidative cleavage and cyclization cascades of N-aryl peptides have been achieved under palladium catalysis with air as the sole stoichiometric oxidant. PMID- 18389131 TI - Indium tribromide-promoted arene-terminated epoxy olefin cyclization. AB - An arene-terminated epoxy olefin cyclization was promoted by a water-tolerant Lewis acid to give tri- and tetracyclic 3beta-hydroxy terpenoids and steroid derivatives in 57 and 37% yields, respectively, per new formed ring up to 75%. PMID- 18389132 TI - Sulfur isotope fractionation during incorporation of sulfur nucleophiles into organic compounds. AB - (34)S enrichment is shown to occur during sulfurization reactions and for the first time conclusively attributed to an isotope equilibrium effect rather than selective addition of (34)S enriched nucleophiles. PMID- 18389133 TI - Supramolecular activation in triggered cascade inversion. AB - An unexpected activation effect from combinations of anionic reagent and amine base resulted in dramatic rate enhancements in multiple carbohydrate cascade inversion. PMID- 18389134 TI - Editorial: Synthesis themed issue. AB - With this issue, NPR fully embraces the molecular and structural biology revolution by showcasing leading work in natural product synthesis, with emphasis on those areas allied to the chemical biology agenda. PMID- 18389136 TI - Yatakemycin: total synthesis, DNA alkylation, and biological properties. AB - DNA-binding small molecules are an important source of anticancer therapeutics that display a diverse array of mechanisms of action. Synthetic studies on the new DNA-alkylating natural product yatakemycin, detailed in this Highlight, have served to reassign its structure, assign the absolute stereochemistry, and provide access to yatakemycin and a series of structural analogues for biological evaluation. Studies on the DNA alkylation properties of (+)-and ent-(-) yatakemycin and related analogues have demonstrated the enhanced DNA alkylation properties of this class of agents and provided insight into their interaction with DNA. PMID- 18389137 TI - The synthetic challenge of diazonamide A, a macrocyclic indole bis-oxazole marine natural product. AB - The complex structure of the marine metabolic diazonamide A comprises a dichlorinated indole bis-oxazole heteroaromatic fragment, and a [b]-fused dihydrobenzofuran-dihydroindole unit containing an animal carbon, all incorporated within a strained double macrocyclic array. This review details the synthetic studies on this fascinating natural product starting from early studies on the original structure (1991-2001), through the synthesis of the originally proposed structure and the subsequent structural revision, to the eventual successful syntheses of the natural product itself. Throughout we focus on the innovative ways in which synthetic chemists have approached the challenges posed by this natural product. PMID- 18389138 TI - Recent advances in biomimetic natural product synthesis. AB - This review highlights some of the most elegant and instructive biomimetic syntheses of natural products over the last few years, providing an updated overview of this area of research. PMID- 18389139 TI - The chemistry of marine furanocembranoids, pseudopteranes, gersolanes, and related natural products. AB - An overview of the chemistry and biology of the diterpene natural products known as the furanocembranoids, pseudopteranes, and gersolanes is provided; 85 references are cited. PMID- 18389140 TI - Recent advances in the total synthesis of pharmaceutically relevant diterpenes. AB - A detailed overview is presented of recent total synthesis approaches to four different diterpenoids--the ginkgolides, tonantzitlolone, ingenol and eleutherobin. Due to their structural complexity, all pose major challenges for total synthesis, guaranteeing diverse strategic tours de force and a rich synthetic chemistry. Additionally, all four diterpenoids are of major relevance because of their biological (target) or pharmaceutical properties. PMID- 18389141 TI - Development of practical syntheses of the marine anticancer agents discodermolide and dictyostatin. AB - Initially isolated in trace quantities from deep-sea sponges, the structurally related polyketides discodermolide and dictyostatin share the same microtubule stabilizing antimitotic mechanism as Taxol. Discodermolide has been the focus of intense research activity in order to develop a practical supply route, and these efforts ultimately allowed its large-scale synthesis and the initiation of clinical trials as a novel anticancer drug. Similarly, the re-isolation and synthesis of dictyostatin continues to stimulate the biological and chemical communities in their quest for the development of new chemotherapeutic agents. This comprehensive review chronicles the synthetic endeavours undertaken over the last 15 years towards the development and realization of practical chemical syntheses of discodermolide and, more recently, dictyostatin, focusing on the methods and strategies employed for achieving overall stereocontrol and key fragment unions, as well as the design and synthesis of novel hybrid structures. PMID- 18389142 TI - Pyranonaphthoquinones--isolation, biological activity and synthesis. AB - A review of the isolation, biological activity and synthesis of pyranonaphthoquinones and closely related compounds is provided. PMID- 18389143 TI - Convergent strategies for the total synthesis of polycyclic ether marine metabolites. AB - Marine polycyclic ether natural products continue to fascinate chemists and biologists due to their exceptionally large and complex molecular architectures and potent and diverse biological activities. Tremendous progress has been made over the past decade toward the total synthesis of marine polycyclic ether natural products. In this area, a convergent strategy for assembling small fragments into an entire molecule always plays a key role in successful total synthesis. This review describes our efforts to develop convergent strategies for the synthesis of polycyclic ethers and their application to the total synthesis of gambierol, gymnocin-A, and brevenal, and to the partial synthesis of the central part of ciguatoxins and the nonacyclic polyether skeleton of gambieric acids. PMID- 18389144 TI - ALA and its clinical impact, from bench to bedside. AB - ALA-induced protoporphyrin IX (PpIX) is used for fluorescence diagnosis (ALA-FD) and for fluorescence-guided resection of both (pre)malignant and non-malignant diseases. ALA is also applied in photodynamic therapy (ALA-PDT) of superficial (pre)malignant lesions in dermatology, urology, neurosurgery, otorhinolaryngology, gynecology and gastroenterology. Today, ALA is approved as Levulan for actinic keratoses, the ALA-methyl ester Metvix for actinic keratoses and basal cell carcinoma, the ALA-hexyl ester Hexvix for the diagnosis of bladder cancer and Gliolan for malignant glioma. The use of ALA for PDT and FD was established around 25 years ago, with most of the fundamental knowledge gained at the "bench" and implemented at the "bedside" due to the diligence of a few researchers within the first 10 years of research. After 1993 ALA research was taken up by many groups. For patient treatment, several factors are relevant. Administered mainly in a topical or oral form, ALA penetrates tissue in a sub optimal way, which is currently improved by special techniques and the use of ALA esters. PpIX accumulation is elevated in many malignant tissues, several tissue abnormalities, and in mucosa. It is also found at elevated levels in macrophages, dendritic cells and activated lymphocytes. Following sufficient PpIX accumulation in the target cells, irradiation is carried out which may be accompanied by a burning sensation at the treatment site. Due to a saturation process of PpIX formation and rapid photobleaching during irradiation the risk of overtreatment is relatively low. Pharmacokinetical studies have demonstrated a low systemic photosensitivity and excretion of PpIX via natural routes. PMID- 18389145 TI - In vivo and in vitro characterisation of a protoporphyrin IX-cyclic RGD peptide conjugate for use in photodynamic therapy. AB - Increasing treatment specificity is one of the major aims of cancer research. Photodynamic therapy is a clinically proven treatment for some cancers and certain other diseases. Photosensitisers generally have little intrinsic selectivity for tumours and any accumulation is dependent upon the type of tumour involved. Increasing tumour selective accumulation could improve the efficacy of PDT and reduce any risk of side effects caused by photosensitiser accumulation in non-target tissue. In order to target photosensitisers to tumours, a cyclic peptide, cRGDfK (arginine-glycine-aspartic acid-phenylalanine-lysine) has been synthesised using solid phase peptide chemistry and conjugated to the porphyrin photosensitiser, protoporphyrin IX. The arginine-glycine-aspartic acid (RGD) motif has been shown to specifically bind alphavbeta3 integrins, heterodimeric glycoproteins upregulated on the surface of proliferating endothelial cells such as those in tumour neovasculature. This study reports the synthesis, in vitro and in vivo characterisation of this novel compound and compares its properties to the free photosensitiser. The individual components in our system, protoporphyrin IX and cRGDfK retain their respective photodynamic and integrin binding activity following the coupling step and produce a conjugate of high purity. The PpIX:cRGDfK conjugate is shown to be a good photosensitiser in vitro in the integrin positive human SiHa cell line and in vivo in a mouse CaNT tumour model. Moreover, pharmacokinetic analysis of PpIX:cRGDfK treated mice shows significant retention and accumulation of photosensitiser in tumour tissue with higher tumour : normal tissue ratios than the free photosensitiser. However, although the conjugate shows this higher accumulation and improved tumour : non-target tissue ratios, the overall in vivo PDT effect, between dose-light intervals of 0 and 6 h, is not significantly better than for free protoporphyrin IX This is possibly due to differences in the target environment or in the subcellular localisation of the compounds. PMID- 18389146 TI - Photophysical properties of dibenzofluorescein and the presence of its tautomers or prototropic forms in organic solvents. AB - The UV/Vis absorption and fluorescence properties of dibenzofluorescein (DBFL) in organic solvents were measured and used to shed light on the possible presence of its tautomers or various prototropic forms. DBFL in aprotic solvents mainly exists in two tautomeric forms, viz. quinoid and lactone, but neither are efficiently fluorescent. In protic solvents, such as methanol and ethanol, both the monoanion and neutral quinoid are present and showed the highest fluorescence quantum yield. In contrast, DBFL is fully dissociated to the monoanion and dianion in deionized water. PMID- 18389147 TI - Microwave discharge electrodeless lamps (MDEL). III. A novel tungsten-triggered MDEL device emitting VUV and UVC radiation for use in wastewater treatment. AB - Exposure to low doses of the xenoestrogen bisphenol A (BPA) and to the hormonal 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide, an environmental endocrine disruptor, can have serious health consequences such as the induction of mammary gland ductal hyperplasias and carcinoma (LaChapelle et al., Reprod. Toxicol., 2007, 23, 20; Murray et al., Reprod. Toxicol., 2007, 23, 383). To the extent that these toxins are present in wastewaters (Donald et al., Sci. Total Environ. 1999, 231, 173; Brotons et al., Environ. Health Perspect. 1994, 103, 608; Olea et al., Environ. Health Perspect. 1996, 104, 298; Biles et al., J. Agric. Food Chem. 1997, 45, 3541; Markey et al., J. Steroid Biochem. Mol. Biol., 2003, 83, 235), we examined their oxidative destruction in aqueous media by a novel light source. A tungsten-triggered microwave discharge electrodeless lamp (W-MDEL) was fabricated for possible use in wastewater treatment using vacuum UV-transparent quartz in which a tungsten trigger, also embedded in quartz, was attached to the MDEL to aid in the self-ignition of the lamp on irradiation at low microwave power levels. The quantity of mercury gas in the W-MDEL was optimized by monitoring the continuous radiation and peak intensities of the emitted light in the vacuum UV (VUV) and UVC regions. The usefulness of the W-MDEL device was assessed through the degradation of 2,4-D and BPA in air-equilibrated aqueous media and in oxygen saturated aqueous media. Enhanced degradation of these two xenoestrogenic toxins was achieved by increasing the number of W-MDEL devices while keeping constant the microwave radiation feeding each W-MDEL lamp. This novel lamp provides an additional light source in the photooxidation of environmental contaminants without the need for a metal-oxide photocatalyst. Under our conditions, process dynamics using the W-MDEL light source are greater than with the more conventional photochemical methods that employ low-pressure Hg arc electrode lamps in synthetic quartz to degrade these two toxic contaminants. PMID- 18389148 TI - UVB sensitivity and cyclobutane pyrimidine dimer (CPD) photolyase genotypes in cultivated and wild rice species. AB - We investigated the UVB-sensitivity in 12 rice strains belonging to two cultivated species (O. sativa and O. glaberrima) and three wild species (O. barthii, O. meridionalis and O. rufipogon) of rice possessing the AA genome, while focusing on the CPD photolyase activity and the genotypes of CPD photolyase. Although the UVB sensitivity, CPD photolyase activity, and CPD photolyase genotype varied widely among these rice species, the sensitivity to UVB radiation depended on the activity of the CPD photolyase, regardless of grass shape, habitat, or species. The rice strains examined here clearly divided into three groups based on the CPD photolyase activity, and the activity of the strains greatly depended on amino acid residues at positions 126 and 296, with the exception of the W1299 strain (O. meridionalis). The amino acid residues 126 and 296 of CPD photolyase in Sasanishiki strain (O. sativa), which showed higher enzymatic activity and more resistance to UVB, were glutamine (Gln) and Gln, respectively. An amino acid change at position 126 from Gln to arginine ("Nori" type) in the photolyase led to a reduction of enzymatic activity. Additionally, an amino acid change at position 296 from Gln to histidine led to a further reduction in activity. The activity of the W1299 strain, which possesses a "Nori" type CPD photolyase, was the highest among the strains examined here, and was similar to that of the Sasanishiki. The CPD photolyase of the W1299 contains ten amino acid substitutions, compared to Sasanishiki. The alterations in amino acid residues in the W1299 CPD photolyase compensated for the reduction in activity caused by the amino acid substitutions at positions 126. Knowledge of the activity of different CPD photolyase genotypes will be useful in developing improved rice cultivars. PMID- 18389149 TI - Transformation of phenolic compounds upon UVA irradiation of anthraquinone-2 sulfonate. AB - This paper studies the transformation of phenol and of 3,5-dichlorophenol (3,5 DCP) upon UVA irradiation of anthraquinone-2-sulfonate (AQ2S). Light-excited AQ2S is able to oxidise phenol, 3,5-DCP, and AQ2S. Transformation reactions do not proceed at a significant extent in the absence of molecular oxygen, in which case recombination reactions of initially formed oxidised and reduced radical species (and/or radical ions) would yield back the initial substrates. AQ2S hydroxyderivatives are the main transformation intermediates, while the phenoxyl radicals arising upon oxidation of phenol and of 3,5-DCP react with the substrates to yield dihydroxybiphenyls and phenoxyphenols. Very small amounts of catechol and of 3,5-dichlorocatechol were observed, indicating a possible minor role of the hydroxyl radicals in the reactivity of the system. Interesting results from an environmental point of view are the formation of 2 hydroxydibenzofuran from phenol and of various tetrachlorinated dihydroxybiphenyls and phenoxyphenols from 3,5-DCP, suggesting that quinone photochemistry can be an important pathway for the formation of hazardous secondary pollutants in the environment. PMID- 18389150 TI - Photoreactivity of indirubin derivatives. AB - Twenty-nine analogs of indirubin, an isomer of indigo, have been synthesized to optimize its promising kinase inhibitory scaffold. These compounds being also pigmented, have been tested for their photoreactivity. Absorption maxima were between 485 nm and 560 nm. Addition of fetal calf serum induced fluorescence and time dependent absorption modifications. Appropriate illumination induced Reactive Oxygen Species (ROS) production for nineteen compounds out of twenty nine. The relationship between fluorescence and ROS production is discussed. Six compounds showed an important toxicity on F98 cells, a murine glioma cell line. Three of these were found to be also phototoxic, as four other non-toxic compounds. All but one phototoxic compounds were detected as ROS producers by in vitro tests. Photoreactivity assessment is important to anticipate adverse reactions for compounds that might be clinically developed. The experimental assay was found to be the only way to evaluate the photoreactivity of this family of compounds since no predictive criteria on structures could be found. Combining the vascular tumor growth inhibition induced by kinase inhibitors with the massive local blood flow arrest following photodynamic treatment may be an efficient anti-cancer strategy. These data could orientate further syntheses of either non-photoreactive compounds or compounds displaying both kinase inhibitory activity and strong phototoxicity. PMID- 18389151 TI - Photocatalysis by tetraphenylporphyrin of the decomposition of chloroform. AB - Irradiation of solutions of tetraphenylporphyrin (H2TPP) in chloroform causes decomposition of the chloroform at UV wavelengths higher than those that decompose chloroform directly. The catalytic cycle involves photooxidation of the porphyrin followed by thermal reduction. Photocatalysis continues after H2TPP has been completely protonated by the HCl produced during decomposition, and the rate of HCl production accelerates as a second pathway, in which CCl3OOH oxidizes the porphyrin, becomes important. PMID- 18389152 TI - Hydrophobicity, topography in membranes and photosensitization of silicon phthalocyanines with axial ligands of varying lengths. AB - Six amphiphilic silicon phthalocyanines (SiPc's) axially linked to a dimethylated amino alkyl group of varying length have been examined for their potential suitability as photosensitizers for photodynamic therapy (PDT). This group of molecules was chosen because the length of the axial ligand might place the chromophoric part of the molecule at different vertical depths in the membrane and possibly affect the extent of membrane localized damage caused by singlet oxygen. We tested the relative penetration depth of the SiPc groups in the membrane by the extent to which their fluorescence was quenched by external iodide ions. We also measured singlet oxygen quantum yields of the SiPc's in a liposome membrane, using the fluorescent target for singlet oxygen, 9,10 dimethylanthracene. The hydrophobicity parameters, LogP, were calculated and were also measured. Some correlation was found between them and Kb's, the binding constants for liposomes. The effect of the axial ligand's length is less striking than in similar cases with hematoporphyrins and protoporphyrins. We link this smaller effect with a bending of the linker chain that enables, sterically, a better positioning of the sensitizer molecules within the ordered lipid layer structure. PMID- 18389153 TI - Rapid determination of the damage to photosynthesis caused by salt and osmotic stresses using delayed fluorescence of chloroplasts. AB - Chloroplasts are one of the most susceptible systems to salt and osmotic stresses. Based on quantitative measurements of delayed fluorescence (DF) of the chloroplasts, we have investigated the damage to photosynthesis caused by these two kinds of stresses in Arabidopsis seedlings by using a custom-built multi channel biosensor. Results showed that the DF intensity and net photosynthesis rate (Pn) decreased in a similar way with increasing NaCl or sorbitol concentration. Incubation of the seedlings in 200 mM NaCl induced a rapid and reversible decline and subsequent slow and irreversible loss in both the DF intensity and Pn. The rapid decline was dominantly related to osmotic stress, whereas the slow declines in the DF intensity and Pn were specific to ionic stress and could be reversed to a similar extent by a Na+-channel blocker. The DF intensity and Pn also exhibited a similar response to irradiation light under NaCl or sorbitol stress. All results indicated that the DF intensity correlated well with Pn under salt and osmotic stresses. We thus conclude that DF is an excellent marker for detecting the damage to photosynthesis caused by these two stresses. The mechanism of the correlation between the DF intensity and Pn under salt and osmotic stresses was also analyzed in theory and investigated with experiments by measuring intercellular CO2 concetration (Ci), stomatal conductance (Gs), chlorophyll fluorescence parameter, and chlorophyll content. This proposed DF technique holds the potential to be a useful means for analyzing the dynamics of salt and osmotic stresses in vivo and elucidating the mechanism by which plants respond to stress. PMID- 18389154 TI - Temperature influence on the energy of nonspecific and specific interactions taking place between 4-aminophthalimide (4-AP) and homogeneous solvents. AB - Temperature induced spectral shifts of the 4-aminophthalimide (4-AP) emission spectra have been measured and compared to the predictions of the McRae solvent induced shift theory (J. Phys. Chem., 1957, 61, 562-572). Three moderately polar chloroalkanes selected as nonspecifically interacting media, and six hydrogen accepting or/and electron pair donating solvents have been used as the media in which the temperature influence on 4-AP-solvent interactions has been studied in the range of 180-320 K. Using the ab initio determined 4-AP ground state dipole moment and fitting appropriate expression originating from the mentioned theory to the shifts found in the chloroalkanes it has been possible to estimate the 4 AP excited state dipole moment, the probe excited state Onsager radius and its gas phase emission spectrum position. Using these values the thermochromic shifts of 4-AP emission spectra in hydrogen bond forming solvents have been predicted and compared to the experimental one. Temperature has been found to have different impact on the changes, upon excitation of the probe, in the mean values of the energies of different hydrogen bonds formed by 4-AP with solvents molecules. PMID- 18389155 TI - Oxygen uptake induced by electron transfer from donors to the triplet state of methylene blue and xanthene dyes in air-saturated aqueous solution. AB - The effects of oxygen in the photolysis of rose bengal, eosin, erythrosin and methylene blue were studied in the presence of formate and electron donors, such as ascorbic acid, aromatic amino acids or aliphatic amines, e.g. triethylamine (TEA). The overall reaction is conversion of oxygen via the hydroperoxyl/superoxide ion radical into hydrogen peroxide. The quantum yield of oxygen uptake (Phi(-O2)) increases with the donor concentration. The photoinduced formation of H2O2 is initiated by quenching of the triplet state of the dye by the donor and subsequent reactions of both the dye and donor radicals with oxygen. For methylene blue and the xanthene dyes in the presence of 10 mM ascorbic acid or 0.1 M TEA Phi(-O2)=0.07-0.25. The spectral and kinetic properties of the specific dye transients, including the radicals involved and the pH and concentration dependences, are discussed. PMID- 18389156 TI - [Prescription tips: ready-made dermatologic agents in individual prescriptions]. PMID- 18389157 TI - [Atopic dermatitis and stress? How do emotions come into skin?]. AB - It is widely accepted, that stress can induce or exacerbate atopic dermatitis. The physiological mechanisms that mediate this negative influence of stress on atopic dermatitis are not clearly understood. This topic has been actively investigated in recent years focusing on neuroimmunological, psychoendocrinological studies and examination of integrity and function of skin barrier under stress. Different neuropeptides and neurotrophins seem to play an important role in stress-induced neurogenic inflammation and connection of nervous and immune system. Mast cells play a key role in the development of inflammatory reaction to stress. Skin barrier is altered by stress by means of increased cortisol level. Thereby lamellar body secretion is decreased and epidermal expression of antimicrobial peptides (beta-defensin and cathelicidin) is down-regulated. We review recent investigations in this field. PMID- 18389158 TI - [Diagnosis of contact allergy. Why in vitro test systems?]. PMID- 18389159 TI - Changes in the sediment trace metal contamination after the commissioning of a municipal wastewater treatment plant in the Souss estuary (South Morocco). AB - Monthly samples of sediment were collected from the Souss estuary, to determine the changes which have occurred in the concentrations of ten metals between 2001 when the low estuary was receiving the wastewaters from Agadir city and 2003 after stopping the sewage dumping. The annual contents of Cu, Fe, Zn (in 2001) and those of Cr, Cu, Fe, Mn, Ni, Pb, and Zn (in 2003) showed significant correlations with those of Al, indicating that fluctuations of their concentrations were only due to those of the aluminosilicate fraction of the clayey sediment. After normalization of these metallic concentrations in relation to those of Al, the contents of the different metals showed insignificant correlations between both years, except for Zn which significantly dropped in 2003 and Cu which increased in 2003. The observed metallic fluctuations are more influenced by the natural conditions of the estuary than the anthropogenic ones. PMID- 18389160 TI - Effect of light on the degradation of two neonicotinoids viz acetamiprid and thiacloprid in soil. AB - Persistence of two neonicotinoids viz thiacloprid and acetamiprid in soil as affected by UV and sunlight exposure was studied. Treated soil was placed in petri-plates, brought to field capacity moisture and then exposed to UV and sunlight. Dissipation for both the pesticides followed monophasic first order kinetics under sunlight, however under UV-light biphasic dissipation was recorded. Residues of acetamiprid and thiacloprid in soil dissipated with half lives of 11.1 and 12.8 days under UV light and 25.1 and 19.1 days under sunlight, respectively. Residues of both the neonicotinoids dissipated quickly under UV light as compared to sunlight. Exposure of thin film of acetamiprid and thiacloprid to UV and sunlight revealed that acetamiprid is more photo labile than thiacloprid. More than 95% acetamiprid dissipated within 24 h as compared to approximately 70% dissipation observed for thiacloprid. PMID- 18389161 TI - Genotoxic effects and LC50 value of NaOCl on Orthrias angorae (Steindachner 1897). AB - Studies show that different organisms used as bio-indicators have indicated several genotoxic and mutagenic effects of disinfected waters. In this study, the 96 h LC(50 )mean value of NaOCl for Orthrias angorae was calculated to be 0.5509 mg/L. The results showed that NaOCl is highly toxic to O. angorae specimens. Statistical analysis demonstrated a significant increase in micronuclei after the induction of 0.5 mg/L NaOCl concentration after 36 h. The same increase has been reported for 0.37 and 0.5 mg/L NaOCl concentrations after 72 h. Even though the MN frequency of 0.37 mg/L was similar after 36 and 72 h, only 72 h micronuclei frequency was statistically significant. The 72 h MN frequency of the negative control group was smaller than 36 h MN frequency of the negative control group. This discrepancy has led to 72 h MN frequency being statistically significant. MN frequency of 0.25 mg/L NaOCl concentration was insignificant when compared to negative test groups. The benzene treatment also caused a significant increase (p < 0.01) in the frequency of micronucleated erythrocytes. PMID- 18389162 TI - Occurrence and downslope mobilization of quaternary herbicide residues in vineyard-devoted soils. AB - The aim of this study was to examine the fate of quaternary herbicides in soil deposits derived from erosion of vineyard-devoted soils. Herbicide occurrence in the crop soils was due to the farmer application inputs. Special attention should be paid to the presence of paraquat (PQ) in the studied vineyard-devoted soils. The major factor governing the sorption of PQ was the solid state organic fraction with the clay mineral content also making a significant contribution. The mobilization and transport of quats-enriched soil particles may generate downslope and downstream environmental problems that mainly affect detritivore ecology. PMID- 18389163 TI - Aqueous solubility, n-octanol-water partition coefficient, and sorption of five selective serotonin reuptake inhibitors to sediments and soils. AB - Aqueous solubilities (S (w)) and n-octanol-water partition coefficients (K (ow)) of five selective serotonin reuptake inhibitors (SSRIs) were measured and sorption to two sediments and three soils with organic matter contents ranging from 0.16% to 1.77% and pH ranging between 5.0 and 7.8 was investigated using a batch equilibrium method. SSRIs had high S (w) (3,022-15,460 mg/l) and relatively low log K (ow) (1.12-1.39). Sorption isotherms followed the Freundlich equation. All SSRIs had sorption capacities of greater than 91% except fluvoxamine with a minimum capacity of 73%. Organic matter contents partly affected sorption, however no correlation between sorption characteristics and cation exchange capacity (CEC) or clay content was observed for any SSRI or adsorbent. Values of K (f), K (d), and log K (oc) ranged from 39 to 18,342, from 60 to 42,579, and 3.35 to 6.02 for the SSRIs. SSRIs likely exhibit mixed mechanisms of sorption such as ionic binding in addition to hydrophobic interactions. PMID- 18389164 TI - Fungal bioremediation of creosote-treated wood: a laboratory scale study on creosote components degradation by Pleurotus ostreatus mycelium. AB - A bioremediation system for creosote-treated wood is proposed, based on the detoxifying capability of Pleurotus ostreatus, a ligninolythic fungus. Non sterilized chipped contaminated wood was mixed at various ratios with wheat straw on which Pleurotus mycelia was grown. At 1:2 initial ratio contaminated wood:wheat straw, chemical analyses demonstrated an almost complete degradation of creosote oil components after 44 days, also confirmed by a significant reduction of ecotoxicity. Lower ratios, i.e. higher amount of contaminated wood, lower system efficiency, although a better creosote degradation was obtained by a stepped up wood addition. PMID- 18389165 TI - Revival of an old method with new techniques: balloon aortic valvuloplasty of the calcified aortic stenosis in the elderly. AB - Balloon aortic valvuloplasty (BAV), introduced since almost 20 years, has experienced a revival for its use in the treatment of elderly patients with severe calcified aortic stenosis that are associated with high operative risk and co-morbidities. This is due to the introduction of new balloon catheters and techniques. This study reports about 75 such cases performed within the past 28 months. The mean age of our patient group was 78 +/- 7 years (median = 80 years). Risk calculation with the EuroSCORE demonstrated an average value of 24.4 +/- 19.5%. BAV was performed along with burst pacing to reduce transvalvular blood flow for stabilization of the balloon catheter until blood pressure dropped to less than 50 mmHg. BAV was performed in 72 patients with a procedural success rate of 73%. There was a decrease of 31 mmHg peak-to-peak gradient across the aortic valve from 63 +/- 35 to 32 +/- 22 mmHg (P < 0.0001). Mean gradient was reduced from 51 +/- 24 to 27 +/- 15 mmHg (P < 0.0001). Aortic valve area increased by 49% from 0.84 +/- 0.33 to 1.25 +/- 0.45 cm(2) (P < 0.0001). Serious adverse events (SAE) occurred in 17% of the 75 BAV procedures. Follow-up revealed a significant improvement in 6-month and 1-year survival. The improved technology of BAV makes this technique attractive for elderly patients who are at high operative risk or in cases where valve replacement was refused for any reason. PMID- 18389166 TI - Spasticity treatment with botulinum toxins. AB - Spasticity is a physiological consequence of an insult to the brain or spinal cord, which can lead to life-threatening, disabling and costly consequences. This typically occurs following stroke, brain injury, spinal cord injury, multiple sclerosis and other disabling neurological diseases and cerebral palsy. It is but one feature of the upper motor neurone syndrome and there have been considerable developments in its management through new drugs and technology. The sole indication for treating spasticity is when it is causing harm and interferes with active or passive functioning. Successful treatment strategies have now been developed and there is good evidence of treatment effectiveness. Treatment is essentially aimed at reducing abnormal sensory inputs, which have an impact on excessive and uncontrolled alpha-motor neuron activity. Attending to the physical characteristics of muscle shortening is the basis of spasticity management. All pharmacological interventions are adjunctive to a programme of physical intervention and there is a good evidence base for this in relation to botulinum toxin treatment. Management therefore centres around the development of a formal treatment plan is important to document the intended outcomes, which should be written and agreed upon with the patient. Anti-spastic drugs treat spasticity. They do not treat contractures and they will not make hemiplegic limbs function, unless the patient's function is impeded by the spasticity. The management of spasticity is physical and all pharmacological interventions are adjunctive to that. This article therefore deals with the principles of management of spasticity and treatment with botulinum toxin. It covers treatment planning, patient assessment, goal setting and covers the range of available treatments. It also describes how botulinum toxin works, the evidence for its use in spasticity management and practical aspects of treatment, such as muscle location, the injection procedure and post-injection care. Finally, there is a word on the organisation of services. The contribution of botulinum toxin to spasticity management is now well recognised. The trick in clinical management is to use it intelligently and to know when and when not to use it. It is a useful short-term means of improving patients' function and the distressing features of spasticity following an insult to the central nervous system. This is usually against the background of a long-term condition, for which a long-term management strategy is required. Botulinum toxin provides a window of opportunity to improve the outcomes from physical management of the focal and multi-focal problems of spasticity. PMID- 18389167 TI - The complete nucleotide sequence and genome organization of bean yellow disorder virus, a new member of the genus Crinivirus. PMID- 18389168 TI - Determination of Nepsilon-(carboxymethyl)lysine in food systems by ultra performance liquid chromatography-mass spectrometry. AB - We report the use of ultra pressure liquid chromatography (UPLC), coupled to a tandem mass spectrometer operated in multiple reaction monitoring mode to determine the advanced glycation endproduct, Nepsilon-(carboxymethyl)lysine (CML). The procedure was applied to acid hydrolyzates of protein isolated from a range of foods (milks processed at different temperatures, butter, cheese, infant formulae, bread, raw and cooked minced beef and olive oil). Highest levels of CML were determined in white bread crust (15.2 +/- 0.63 mmol/mol Lys), wholemeal bread crust (13.1 +/- 0.61 mmol/mol Lys) and evaporated full-fat milk (4.86 +/- 0.77 mmol/mol Lys). Lowest levels of CML were measured in raw minced beef beef (0.03 +/- 002 mmol/mol Lys), raw full-fat cow's milk (0.08 +/- 0.03 mmol/mol Lys) and pasteurized skimmed cow's milk (0.09 +/- 0.002 mmol/mol Lys). CML could not be detected in olive oil. PMID- 18389169 TI - Binding of brilliant red compound to lysozyme: insights into the enzyme toxicity of water-soluble aromatic chemicals. AB - The non-covalent interaction of brilliant red (BR) with lysozyme was investigated by the UV spectrometry, circular dichroism (CD) and isothermal titration calorimetry (ITC). The thermodynamic characterization of the interaction was performed and the assembly complexes were formed: lysozyme(BR)(17) at pH 2.03, lysozyme(BR)(15) at pH 3.25 and lysozyme(BR)(12) at pH 4.35, which corresponded to the physiological acidities. The ionic interaction induces a combination of multiple non-covalent bonds including hydrogen bond, hydrophobic interaction and van der Waals force. The two-step binding model of BR was found, in which one or two BR molecules entered the hydrophobic intracavity of lysozyme and the others bound to the hydrophilic outer surface of lysozyme. Moreover, BR binding resulted in change of the lysozyme conformation and inhibition of the lysozyme activity. The possible binding site and type of BR and the conformational transition of lysozyme were speculated and illustrated. This work provided a useful approach for study on enzyme toxicity of aromatic azo chemicals. PMID- 18389170 TI - Changes in free amino acids in the brain during embryonic development in layer and broiler chickens. AB - Developmental changes in the levels of the excitatory amino acids L-glutamate (Glu) and L-Aspartate (Asp) and inhibitory amino acids glycine (Gly) and gamma amino butyric acid (GABA), as well as taurine and its related amino acids L methionine (Met), L-cysteine (Cys) and L-serine (Ser) in the brain and pectoralis muscle at various embryonic stages and hatch in broiler and layer type chickens were determined. Brain concentrations of Asp, GABA and taurine were higher than those in the muscle, but the difference in the two types was small. The concentrations of the precursors of taurine including Met, Cys and Ser were lower than that of taurine. In conclusion, the synthesis of some amino acids and their metabolites such as Asp, GABA and taurine in the chick embryo is very high in order to support brain development. PMID- 18389171 TI - Local anaesthesia as a risk factor for recurrence after groin hernia repair. AB - BACKGROUND: The literature is inconclusive regarding the effect of local infiltration anaesthesia on the risk of recurrence after groin hernia repair. METHODS: Based upon the Danish Hernia Database, reoperations after a primary Lichtenstein mesh repair were assessed in 43,123 patients between 1998 and 2005. Data were analysed in relation to type of inguinal hernia and whether repaired in public hospitals or by private hernia surgeons. RESULTS: The nationwide data showed no overall difference in reoperation rate (about 3.5% after five years), whether performed in local or general/regional anaesthesia. Reoperation rates were about 50% higher when performed for a direct hernia than for an indirect hernia. The use of local anaesthesia was followed by higher reoperation rates compared with general or regional anaesthesia after repair of a direct but not an indirect hernia in hospitals. However, reoperation rates after both direct and indirect hernia repairs were lower among private hernia surgeons, with uniform use of local anaesthesia compared with reoperations following primary surgery in hospitals. CONCLUSION: These extensive nationwide data suggest that surgical experience and hernia type may be important factors for reoperation, and that it is independent of the type of anaesthesia. When performed in general hospitals, local anaesthesia may be a risk factor for recurrence after primary repair of a direct hernia. PMID- 18389172 TI - Taxonomic status of Monotropastrum humile, with special reference to M. humile var. glaberrimum (Ericaceae, Monotropoideae). AB - Taxonomic treatment of the achlorophyllous monotropoid plant Monotropastrum humile is still unclear and confusing because of the lack of detailed morphological analyses and molecular phylogeny. In particular, the taxonomic status of a glabrous variety, M. humile var. glaberrimum, is under debate. Our detailed examination of the morphological characteristics of living plants revealed that M. humile var. glaberrimum can be easily distinguished from the putative conspecific taxon M. humile var. humile by characteristics not previously recognized, namely the shape and color of the floral disc. Most morphological features characterizing Cheilotheca were also found in M. humile var. glaberrimum. Moreover, there was considerable nucleotide differentiation in the internal transcribed spacer (ITS)2 sequences of M. humile var. humile and var. glaberrimum. Molecular analysis of the phylogenetic relationship of M. humile var. humile, var. glaberrimum, and other monotropoids using ITS2 sequences showed that two varieties of M. humile formed a monophyletic clade with a member of a different genus, Monotropa L., but obvious phylogenetic relationships among these three taxa were not obtained. Thus we conclude that Monotropastrum humile var. glaberrimum should be treated as a distinct species. However, the generic affiliation of M. humile var. glaberrimum could not be determined because of its intermediate character state combination and the insufficient characterization of related species. We strongly suggest that Monotropastrum as a whole needs re evaluation. PMID- 18389173 TI - Biomass production, photosynthesis, and leaf water relations of Spartina alterniflora under moderate water stress. AB - The perennial smooth cordgrass, Spartina alterniflora, has been successfully introduced in salty ecosystems for revegetation or agricultural use. However, it remains unclear whether it can be introduced in arid ecosystems. The aim of this study was to investigate the physiological response of this species to water deficiency in a climate-controlled greenhouse. The experiment consisted of two levels of irrigation modes, 100 and 50% field capacities (FC). Although growth, photosynthesis, and stomatal conductance of plants with 50% FC were reduced at 90 days from the start of the experiment, all of the plants survived. The water stressed plants exhibited osmotic adjustment and an increase in the maximum elastic modulus that is assumed to be effective to enhance the driving force for water extraction from the soil with small leaf water loss. An increase in the water use efficiency was also found in the water-stressed plants, which could contribute to the maintenance of leaf water status under drought conditions. It can be concluded that S. alterniflora has the capacity to maintain leaf water status and thus survive in arid environment. PMID- 18389174 TI - Long term follow-up of a membranous glomerulopathy as late complication of chronic graft-versus-host disease following allogeneic stem cell transplantation. PMID- 18389175 TI - Actual status of AIDS-related lymphoma management in Japan. PMID- 18389176 TI - A retrospective study of the safety of BCNU wafers with concurrent temozolomide and radiotherapy and adjuvant temozolomide for newly diagnosed glioblastoma patients. AB - Despite aggressive therapy, most patients with glioblastoma multiforme (GBM) die within 2 years of diagnosis. The efficacy and safety of carmustine (BCNU) wafers followed by radiotherapy have been demonstrated in patients with malignant glioma. However, there is a reluctance to recommend them for newly diagnosed GBM patients due to the potential toxicity of BCNU wafers combined with temozolomide (TMZ) chemotherapy and radiotherapy. The purpose of this study was to assess the safety of BCNU wafers implanted at initial surgery, followed by concurrent TMZ and radiotherapy, and then adjuvant TMZ for the treatment of newly diagnosed GBM. We conducted a retrospective analysis of clinic and hospital records of 21 newly diagnosed GBM patients who received multimodal therapy at Florida Hospital Cancer Institute from January 2003 to December 2005. Three of 21 patients had grade 3 toxicities (two with cerebritis, one with psychosis). Grade 4 toxicities were not observed. Median overall survival was 17 months, median progression-free survival was 8.5 months, and 2-year survival was 39%. Multimodal treatment with surgery, BCNU wafers, radiotherapy, and TMZ did not result in a notable increase in significant toxicities. Survival outcomes were comparable to those in other studies in which patients were treated with concurrent TMZ and radiotherapy followed by adjuvant TMZ. Thus, the implantation of BCNU wafers prior to TMZ and radiotherapy appears safe in newly diagnosed GBM patients. PMID- 18389177 TI - Time course of imaging changes of GBM during extended bevacizumab treatment. AB - Glioblastoma multiforme (GBM) are morphologically heterogeneous tumors, with varying amounts of necrosis, and edema. Previous studies have shown that treatments incorporating the VEGF antibody bevacizumab can reduce edema and tumor burden in GBM. Additionally it has been suggested that bevacizumab regimen treatment reduces the percent of tumoral necrosis. Therefore we sought to (1) determine the time course of change in necrosis, tumor, and edema volume in patients who respond to bevacizumab regimen treatment and (2) determine if GBM that progress following a response to bevacizumab regimen treatment are morphologically different from their appearance at prior tumor progression. Therefore, we retrospectively assessed tumor, necrosis, and edema volumes on MRI scans from 15 patients with recurrent GBM who responded to bevacizumab regimen treatment, and had extended (>7 month) follow-up. We found that the median time to best tumor response was 158 days (range, 16-261, SD = 63). The median best response was 72.1% reduction in tumor volume and 72.8% reduction in peritumoral edema. Most tumors (77.8%) showed resolution of necrotic areas. The relative reduction of edema and necrosis was sustained, even in patients (n = 7) who developed tumor progression. Thus the mean ratio of edema-to-tumor volume at progression on bevacizumab regimen treatment was 38.4% lower than that for the same tumors seen on progression scans following prior chemotherapy. The percentage of necrotic tumor also was diminished following progression on bevacizumab regimen treatment. These findings illustrate the time course of changes in edema and tumor volume with prolonged bevacizumab regimen treatment, and support the conclusion that the morphology of recurrent GBM following bevacizumab regimen therapy is distinct from that on other chemotherapy. PMID- 18389178 TI - Management of chronic musculoskeletal pain in the elderly: opinions on oral medication use. AB - The use of oral medication in the treatment of chronic musculoskeletal pain in the elderly requires careful selection of drugs to control pain with consideration for both the physiological state and the presence of disease(s). Recent advances have improved the understanding of biomolecular mechanisms of chronic pain. These include the production of powerful pro-inflammatory cytokines by glial and microglial cells, which then lead to activation of major pain pathways from the periphery through the dorsal horn and supra-spinal pathways to the somatosensory and other higher cortical centres. This has allowed better recognition for intervention with anti-inflammatory agents to control cytokine production (e. g. prednisolone, triamcinolone and other brain-penetrating corticosteroids). Advances in understanding of chronic pain have lead to recognition of neuronal PX2 puringergic receptors as potential sites for drugs to control pain by more selective actions. Pain control in the elderly involves extensive use of analgesics, among them the non-steroidal anti-inflammatory drugs (NSAIDs), paracetamol (acetaminophen), and various narcotics. Each of these has its drawbacks, mostly related to potential toxicities. Attempts to reduce the serious gastro-intestinal (GI) adverse effects of the NSAIDs by the introduction of the highly selective COX-2 inhibitors (coxibs) have only had limited benefit in reducing these untoward actions. Moreover, the risks of serious cardiovascular (CV) and renal side-effects, though statistically infrequent, are none the less of major concern. Cardio-renal effects have been attributed to some (e. g. diclofenac), but not all (e. g. naproxen) conventional NSAIDs. Here we make recommendations for a selection of certain NSAIDs to be used for pain therapy in the elderly in consideration of their relative safety and pharmacokinetics. While newer formulations of narcotics have given some advance in pain control, the application of this group of drugs requires close supervision in the elderly, especially those with cognitive decline, since drug actions on the central and peripheral nervous systems (CNS, PNS) can result in significant adverse effects of these agents (e. g. constipation, drowsiness, respiratory and cardiovascular decline). Improvements in the safety and effectiveness of musculoskeletal pain therapy in the elderly can only be achieved by identification and frequent re evaluation of the cause of the chronic pain and the impact on the patient's general medical state. PMID- 18389179 TI - Persistent parasites and immunologic memory in cutaneous leishmaniasis: implications for vaccine designs and vaccination strategies. AB - Despite a plethora of publications on the murine model of cutaneous leishmaniasis and their contribution to our understanding of the factors that regulate the development of CD4+ T cell immunity in vivo, there is still no effective vaccine against the human disease. While recovery from natural or experimental infection with Leishmania major, the causative agent of human cutaneous leishmaniasis, results in persistence of parasites at the primary infection site and the development of long-lasting immunity to reinfection, vaccination with killed parasites or recombinant proteins induces only short-term protection. The reasons for the difference in protective immunity following recovery from live infection and vaccination with heat-killed parasites are not known. This may in part be related to persistence of live parasites following healing of primary cutaneous lesions, because complete clearance of parasites leads to rapid loss of infection induced immunity. Recent reports indicate that in addition to persistent parasites, IL-10-producing natural regulatory T cells may also play critical roles in the maintenance and loss of infection-induced immunity. This review focuses on current understanding of the factors that regulate the development, maintenance and loss of anti-Leishmania memory responses and highlights the role of persistent parasites and regulatory T cells in this process. Understanding these factors is crucial for designing effective vaccines and vaccination strategies against cutaneous leishmaniasis. PMID- 18389180 TI - Therapeutic hypothermia for acute liver failure: toward a randomized, controlled trial in patients with advanced hepatic encephalopathy. AB - Acute liver failure (ALF), the abrupt loss of liver function in a patient without previous liver disease, remains a highly mortal condition. Patients with ALF often succumb to their liver injury after the development of cerebral edema, resulting in intracranial hypertension and brain herniation. While the management of cerebral edema in ALF always includes the administration of osmotically active agents, osmotherapy often reduces intracranial pressure (ICP) insufficiently, such that herniation may be delayed but not prevented. Therapeutic hypothermia, the intentional reduction of body core temperature, has been increasingly used to treat cerebral edema in patients with traumatic and hypoxic brain injury. Data in animal models of ALF also suggest that hypothermia is effective in the prevention and treatment of cerebral edema, and case reports in humans have suggested that hypothermia is an effective bridge to orthotopic liver transplantation. A randomized, controlled trial comparing the management of ALF patients under normothermic and hypothermic conditions is a logical extension of these preliminary observations. Herein, we consider the many difficulties which will be encountered in the design of such a trial in patients with ALF at high risk of developing cerebral edema. PMID- 18389181 TI - Treatment of a basilar artery occlusion with intra-arterial thrombolysis in a 3 year-old girl. AB - INTRODUCTION: Posterior circulation stroke attributable to basilar artery thrombosis is a rare entity and is difficult to diagnose in young children. Given the high rate of disability and mortality associated with the condition, endovascular revascularization may be a potential treatment strategy. CASE: We describe a 3-year-old girl who presented with a decline in consciousness associated with tonic and clonic movements of her extremities thought to be secondary to a seizure. She was found to have a basilar artery occlusion 18 h from last known to be normal and was successfully treated with intra-arterial alteplase. At 6 month follow up the child has made substantial improvements and is in preschool with difficulties with ambulating longer distances. CONCLUSION: We report the youngest patient in the literature to date who was treated with intra-arterial thrombolysis for an acute basilar artery occlusion. PMID- 18389182 TI - Migration of embolization coils: our experience. PMID- 18389183 TI - Successful embolization of a renal artery pseudoaneurysm with arteriovenous fistula and extravasations using Onyx after partial nephrectomy for renal cell carcinoma. AB - Partial nephrectomy can be associated with vascular complications. Computed tomography (CT) with CT angiography is ideal for noninvasive imaging of this process. The treatment of choice is selective embolization. Successful transcatheter embolization of right renal subsegmental artery pseudoaneurysm with arteriovenous fistula and extravasations using Onyx was performed in a 66-year old woman with macrohematuria 12 days after partial nephrectomy for renal cell carcinoma. PMID- 18389184 TI - Endovascular repair of an actively hemorrhaging stab wound injury to the abdominal aorta. AB - Traumatic injury of the abdominal aorta is rare and potentially lethal (Yeh et al., J Vasc Surg 42(5):1007-1009, 2005; Chicos et al., Chirurgia (Bucur) 102(2):237-240, 2007) as it can result in major retroperitoneal hemorrhage, requiring an urgent open surgery. In case of concomitant bowel injury or other conditions of hostile abdomen, endovascular repair can be an alternative treatment. This case report deals with a 50-year-old man presenting at the emergency ward with three stab wounds: two in the abdomen and one in the chest. During explorative laparotomy, liver laceration and bowel perforation were repaired. One day later, abdominal CT-scan revealed an additional retroperitoneal hematoma associated with an aortic pseudoaneurysm, located anteriorly 3 cm above the aortic bifurcation. Because of the risk of graft infection, an endovascular repair of the aortic injury using a Gore excluder stent-graft was performed. Radiological and clinical follow-up revealed a gradual shrinkage of the pseudo aneurysm and no sign of graft infection at two years' follow-up. PMID- 18389186 TI - Percutaneous vertebroplasty and bone cement leakage: clinical experience with a new high-viscosity bone cement and delivery system for vertebral augmentation in benign and malignant compression fractures. AB - The aim of this study was to assess the feasibility of and venous leakage reduction in percutaneous vertebroplasty (PV) using a new high-viscosity bone cement (PMMA). PV has been used effectively for pain relief in osteoporotic and malignant vertebral fractures. Cement extrusion is a common problem and can lead to complications. Sixty patients (52 female; mean age, 72.2 +/- 7.2) suffering from osteoporosis (46), malignancy (12), and angiomas (2), divided into two groups (A and B), underwent PV on 190 vertebrae (86 dorsal, 104 lumbar). In Group A, PV with high-viscosity PMMA (Confidence, Disc-O-Tech, Israel) was used. This PMMA was injected by a proprietary delivery system, a hydraulic saline-filled screw injector. In Group B, a standard low-viscosity PMMA was used. Postprocedural CT was carried out to detect PMMA leakages and complications. Fisher's exact test and Wilcoxon rank test were used to assess significant differences (p < 0.05) in leakages and to evaluate the clinical outcome. PV was feasible, achieving good clinical outcome (p < 0.0001) without major complications. In Group A, postprocedural CT showed an asymptomatic leak in the venous structures of 8 of 98 (8.2%) treated vertebrae; a discoidal leak occurred in 6 of 98 (6.1%). In Group B, a venous leak was seen in 38 of 92 (41.3%) and a discoidal leak in 12 of 92 (13.0%). Reduction of venous leak obtained by high viscosity PMMA was highly significant (p < 0.0001), whereas this result was not significant (p = 0.14) related to the disc. The high-viscosity PMMA system is safe and effective for clinical use, allowing a significant reduction of extravasation rate and, thus, leakage-related complications. PMID- 18389185 TI - Combined interventional and surgical treatment for acute aortic type a dissection. AB - Surgical repair and endovascular stent-graft placement are both therapies for thoracic aortic dissection. A combination of these two approaches may be effective in patients with type A dissection. In this study, we evaluated the prognosis of this combined technique. From December 2003 to December 2006, 15 patients with type A dissection were admitted to our institute; clinical data were retrospectively reviewed. Follow-up was performed at discharge and approximately 12 months after operation. Endovascular stent-graft placement by interventional radiology and surgical repair for reconstruction of aortic arch was performed in all patients. Total arch replacement for distal arch aneurysm was carried out under deep hypothermia with circulatory arrest; antegrade selected cerebral perfusion was used for brain protection. Four patients concomitantly received a coronary artery bypass graft. Hospital mortality rate was 6.7%; the patient died of cerebral infarction. Neurological complications developed in two patients. Multi-detector-row computed tomography scans performed before discharge revealed complete thrombosis of the false lumen in six patients and partial thrombosis in eight patients. At the follow-up examination, complete thrombosis was found in another three patients, aortic rupture, endoleaks, or migration of the stent-graft was not observed and injuries of peripheral organs or anastomotic endoleaks did not occur. For patients with aortic type A dissection, combining intervention and surgical procedures is feasible, and complete or at least partial thrombosis of the false lumen in the descending aorta can be achieved. This combined approach simplified the surgical procedures and shortened the circulatory arrest time, minimizing the necessity for further aortic operation. PMID- 18389188 TI - Comparison of the clinical efficacy of flutamide and spironolactone plus ethinyloestradiol/cyproterone acetate in the treatment of hirsutism: a randomised controlled study. AB - INTRODUCTION: Hirsutism is commonly a consequence of ovarian androgen over production. Polycystic ovary syndrome (PCOS) or peripheral hypersensitivity to normal androgen circulating levels (idiopathic hirsutism) can be the underlying cause. Several drugs with anti-androgenic properties, such as cyproterone acetate (CPA), spironolactone and flutamide have been used to treat hirsutism, but the efficacy of these drugs has yet to be fully elucidated. The objective of this study was to compare the effectiveness of flutamide, and spironolactone plus a combination tablet of 2 mg CPA/35 microg ethinyloestradiol (EE) in the treatment of hirsutism. METHODS: A prospective randomised clinical study was conducted in a tertiary care hospital setting. Twenty-nine women with hirsutism as a consequence of PCOS or idiopathic hirsutism were randomly assigned to receive 250 mg/day flutamide alone or 100 mg/day spironolactone plus a combination tablet of 2 mg CPA/35 microg EE, for 6 months. Patients' hormonal and lipid profiles were evaluated. Hirsutism was graded according to the modified Ferriman-Gallwey (mF-G) score, and side effects were monitored. RESULTS: A significant decrease in mF-G scores was observed in the flutamide (from 11.2+/-3.3 to 7.6+/-4.0) and spironolactone plus CPA/EE (from 9.9+/-1.9 to 7.1+/-2.0) groups. However, there was no statistically significant difference between the two groups. After flutamide therapy, total cholesterol levels decreased significantly but no significant change was observed in any other lipid parameters or in the patients' hormone profiles. After spironolactone plus CPA/EE therapy, levels of luteinising hormone, total testosterone and free testosterone significantly decreased and triglyceride levels increased. No patients were found to have abnormal liver function test results. CONCLUSION: Flutamide and spironolactone plus CPA/EE are effective drugs in the treatment of hirsutism. PMID- 18389187 TI - Transcatheter arterial chemoembolization (TACE) with lipiodol to treat hepatocellular carcinoma: survey results from the TACE study group of Japan. AB - The purpose of this study was to retrospectively clarify the current status in Japan of TACE using Lipiodol together with anticancer agents to treat hepatocellular carcinoma (HCC). We retrospectively surveyed 4,659 (average annual total) procedures for HCC over the years 2002-2004 at 17 institutions included in the TACE Study Group of Japan. The survey included six questions that were related mainly to TACE and Lipiodol for HCC treatment. The most frequently applied among the 4,659 procedures at the 17 institutions were TACE (2,310; 50%) and local ablation (1,395; 30%). Five of the institutions applied 201-300 procedures and 4 applied 101-200. Lipiodol was used in "all procedures" and in "90% or more" at seven and nine institutions, respectively. Almost all institutions applied 4-6 (mean, 5) ml of Lipiodol during TACE to treat tumors 5 cm in diameter. In conclusion, this survey clarified that TACE using Lipiodol and anticancer agents is a popular option for HCC treatment in Japan. PMID- 18389189 TI - [Diary of a hospital evacuation. Discovery of a 5 hundredweight bomb from World War II]. AB - The discovery of an aircraft bomb from World War II made the complete evacuation of a tertiary care hospital with 629 beds and 17 specialist departments including a neonatal intensive care unit necessary. Some months before an alarm plan had been issued and a fire practice had been carried out which made it obvious to all concerned how important such measures are. Nevertheless, more room for improvement could be learned from the evacuation, in particular the rapid classification of the patients into categories and the fact that 20-30% of the patients needed stretcher-based transport for evacuation. PMID- 18389190 TI - Transient spinal myoclonus after spinal anaesthesia with bupivacaine in the perioperation period. PMID- 18389191 TI - [Cost of intensive care in a German hospital: cost-unit accounting based on the InEK matrix]. AB - OBJECTIVE: The aim of this study was to determine the actual cost per intensive care unit (ICU) day in Germany based on routine data from an electronic patient data management system as well as analysis of cost-driving factors. A differentiation between days with and without mechanical ventilation was performed. METHODS: On the ICU of a German focused-care hospital (896 beds, 12 anesthesiology ICU beds), cost per treatment day was calculated with or without mechanical ventilation from the perspective of the hospital. Costs were derived retrospectively with respect to the period between January and October 2006 by cost-unit accounting based on routine data collected from the ICU patients. Patients with a length of stay of at least 2 days on the ICU were included. Demographic, clinical and economical data were analyzed for patient characterization. RESULTS: Data of 407 patients (217 male and 190 female) were included in the analysis, of which 159 patients (100 male, 59 female) were completely or partially mechanically ventilated. The mean simplified acute physiology (SAPS) II score at the onset of ICU stay was 28.2. Average cost per ICU day was 1,265 EUR and costs for ICU days with and without mechanical ventilation amounted to 1,426 EUR and 1,145 EUR, respectively. Personnel costs (50%) showed the largest cost share followed by drugs plus medicinal products (18%) and infrastructure (16%). CONCLUSIONS: For the first time, a cost analysis of intensive care in Germany was performed with routine data based on the matrix of the institute for reimbursement in hospitals (InEK). The results revealed a higher resource use on the ICU than previously expected. The large share of personnel costs on the ICU was evident but is comparable to other medical departments in the hospital. The need for mechanical ventilation increases the daily costs of resources by approximately 25%. PMID- 18389192 TI - [Emergency medicine--medicine for an ageing society. A contribution to the context of emergency missions for elderly people]. AB - Due to fundamental demographic as well as social changes, the emergency medical services (EMS) have to respond to an increasing number of geriatric emergencies. By means of some typical case histories the practical problems arising in preclinical emergency medical intervention and the central role of context factors like social isolation, reduced mental capabilities and the resulting need for help are demonstrated. It is discussed how emergency medical services (EMS) can contribute to the problems of an ageing society beyond the scope of a system which is dedicated only to the individual. One possibility is the epidemiological analysis of geriatric emergencies, the accompanying context factors and the development of an adequate infrastructure which is adapted to the needs of the elderly. The EU project EMERGE is an example of how emergency medical expertise is utilized in an interdisciplinary cooperation. An automatically working system based on ambient sensor technology is developed for early detection and prevention of emergency situations in the home environment. Supportive technology ("assisted living") should enable the elderly to live a safe and self-determined life as long as possible. Integration of this additional information into the processes of Emergency Medical Services (EMS) is the logistic prerequisite to establish a social medical assistance tailored to the needs of an ageing society. PMID- 18389194 TI - ["Friendship on the border of science and research". Special colloquium on the occasion of the bestowal of the Otto Braun-Falco Stipendiums 2007]. PMID- 18389193 TI - [Fistulas and fissures. Part II: fissures]. AB - Fissures in ano are linear ulcers situated in the anal canal and extending from the dentate line to the margin of the anus. They cause pain and spasms. Diagnosis is made by the history alone. Local medical treatment might consist of topical 0.4% glycerol trinitrate or 2% calcium blocker. In case of therapy resistance, botulinum toxin injection into the internal sphincter is an effective but expensive alternative with encouraging results. If medical treatment fails, then operation has to be recommended. As lateral internal sphincterotomy represents poses a clear danger to continence, fissurectomy combined with the excision of skin tags and any anal papilla is now the operative treatment of choice. PMID- 18389195 TI - [Thrombocytopenia in a patient with coronary artery disease after percutaneous coronary intervention]. AB - Thrombocytopenia in patients with percutanous coronary intervention is a known complication of glycoprotein IIb/IIIa inhibitors. This can limit the application of these agents. Platelet count monitoring 2, 6, 12 and 24 hours after starting the treatment reveals most cases of acute thrombocytopenia. Side effects can be avoided by the early discontinuation of the glycoprotein IIb/IIIa antagonist treatment. A selective diagnostic approach by laboratory measures should exclude any confusion with heparin-induced thrombocytopenia and pseudo thrombocytopenia. PMID- 18389196 TI - [Hormone therapy and anti-aging: is there an indication?]. AB - The desire for a long life is deeply embedded in nearly all men. Fortunately life expectancy has remarkably increased over the past decades, on the other hand advancing age is frequently associated with a rise in morbidity. Above simply prolonging life there is a need to search for strategies to improve the quality of life in the elderly. Different substances to prevent premature aging, cancer and degenerative disorders appear to be promising candidates. Since it has been suggested that the decline of different hormones over the lifespan is closely related to the aging process replacement of these hormones may be a strategy against aging. Especially hormones like growth hormone, DHEA, testosterone and melatonin were considered as anti-aging agents. This review is focusing on the theoretical background and the previously known effects of different hormones to slow aging processes. Despite some promising results in a variety of studies conducted over the past years presently available data do not justify the broad use of hormones for anti-aging purposes. However, although no single hormone can be recognized as a 'rejuvenating' and life extending agent, some of their actions may be beneficial for the aging process. PMID- 18389197 TI - [Preformed plate-fixation system for type AO 13C3 distal humerus fractures: clinical experiences and treatment results taking access into account]. AB - In distal intraarticular humerus fractures primary stable osteosynthesis is essential for early mobilization of the elbow joint. Double-plate osteosynthesis techniques using different configurations are the gold standard. In the literature plate position is sometimes discussed controversially. In cases of distal humerus fractures (type AO 13C3) with metaphyseal comminution, as well as in elderly patients with poor bone quality, utilizing locking plates with angular stability was found to have increased structural properties with regard to primary fixation stability. The dorsal approach with osteotomy of the olecranon seems to be very effective in open reduction and internal fixation of this type of fracture. One new development is the anatomically preformed plate-fixation systems such as the LCP-System for distal humerus (Synthes). This study presents our first experience with this system in 11 cases of open reduction and internal fixation of type AO 13C3 distal humerus fractures. The system-specific features and intraoperative options were analyzed. PMID- 18389198 TI - [The delegation of medical services: is it a Trojan horse?]. AB - Delegation of medical treatment and responsibility from doctors to nonphysicians are being advocated more and more by public health politicians. The opinion of the German Union of Orthopaedic and Trauma Surgery is outlined. Definitions for treatment areas that cannot or may be delegated are presented. Physicians must be spared from overloading of administrative work. Delegating must not result in deterioration of patient care or destruction of the medical profession. Options and risks of delegation are discussed. PMID- 18389199 TI - [The problem of delegating medical activities: a balancing act between ensuring continuing education and relieving the workload in medical service]. PMID- 18389200 TI - [Accidental abdominal needlestick injury incurred while discarding a disposal container]. AB - Needlestick injuries routinely occur in everyday clinical practice. Adequate instruction of employees in health care and correct prophylaxis against exposure could conspicuously reduce the incidence. Successful prevention of chronic infectious diseases comprises strict vaccination plans and substantial knowledge of post-exposure prophylaxis. The introduction of self-securing cannulas and injection instruments represents an important technological advance. PMID- 18389201 TI - Hierarchy length in orphaned colonies of the ant Temnothorax nylanderi. AB - Workers of the ant Temnothorax nylanderi form dominance orders in orphaned colonies in which only one or a few top-ranking workers begin to produce males from unfertilized eggs. Between one and 11 individuals initiated 80% of all aggression in 14 queenless colonies. As predicted from inclusive fitness models (Molet M, van Baalen M, Monnin T, Insectes Soc 52:247-256, 2005), hierarchy length was found to first increase with colony size and then to level off at larger worker numbers. The frequency and skew of aggression decreased with increasing size, indicating that rank orders are less pronounced in larger colonies. PMID- 18389202 TI - [Thrombosis of developmental venous anomaly and consecutive venous infarction]. AB - Developmental venous anomalies (DVA) are congenital aberrations in the cerebral venous system reflecting failure of normal embryogenesis. They are considered the most common intracranial vascular malformation. Although they are generally regarded as benign entities, their clinical significance remains controversial- case reports do exist considering DVA to be the origin of thrombosis, cerebral ischemia, and hemorrhage. We report a 26-year-old patient suffering from thrombosis of a DVA. PMID- 18389203 TI - [Neurolymphomatosis. Subacute sensorimotor polyneuropathy as a first sign of non Hodgkin's B cell lymphoma]. AB - We report on a 65-year-old patient with subacute painful sensorimotor polyneuropathy with distal leg muscle palsy and initially presenting with bilateral leg edema. Electrophysiologic testing showed an axonal lesion pattern with acute denervation. Nerve biopsy demonstrated neurolymphomatosis as an initial manifestation of a non-Hodgkin's B cell lymphoma. PMID- 18389204 TI - [Pregnancy and epilepsy. Retrospective analysis of 118 patients]. AB - BACKGROUND: The aim of this study was to evaluate the treatment of women with epilepsy investigated prior to or during pregnancy. PATIENTS: One hundred eighteen women (median age 28.9 years) with idiopathic (n=43), cryptogenic (n=41), or symptomatic (n=29) epilepsy and appearing for special pregnancy planning advice at a German epileptology clinic were evaluated. All patients were investigated between 2002 and 2007. FINDINGS: Of the study patients, 69 were seen prior to pregnancy, 41 (59.4%) were on monotherapy with antiepileptic drugs (AED), and 22 (31.9%) were already on folic acid supplementation. A change in AED medication was recommended in 50 (72.5%). Ninety-three of the patients were seen during pregnancy, most often during the first trimenon (n=44). Fifty-one (55%) were on AED monotherapy, most often with lamotrigine (n=24) or valproate (n=13). During pregnancy, seizure frequency increased in 33 women (35.5%) and decreased in 14 (15%). CONCLUSION: Monotherapy with AED should be established if possible, and folic acid supplementation should be started prior to pregnancy. PMID- 18389205 TI - [Development of the evidence-based S3 guideline for diagnosis and therapy of bipolar disorders]. PMID- 18389207 TI - [The future of ESWL]. AB - With the introduction of the Dornier HM3 lithotripter, the successful history of extracorporeal shock wave lithotripsy (ESWL) for noninvasive treatment of urinary stones began about 25 years ago. The development of newer lithotripters has not been able to improve clinical efficacy because the shock wave parameters specifically responsible for stone disintegration or tissue trauma and pain induction have not yet been identified. Actual research in lithotripter technology deals with modification of the focal point. The evolution of endoscopic procedures, ureterorenoscopy, and percutaneous nephrolithotomy took longer but was more successful in terms of clinical efficacy. Nowadays, ESWL or an endoscopic procedure is offered as a reasonable option for most urinary stone cases. Therefore, economic aspects and the surgeon's expertise will become greater factors when a procedure is chosen. ESWL, with or without anaesthesia, will be an inherent part of future treatment modalities for urinary stones. PMID- 18389208 TI - Diversity and linkage disequilibrium analysis within a selected set of cultivated tomatoes. AB - Within the Dutch genomics initiative the "Centre for Biosystems Genomics" (CBSG) a major research effort is directed at the identification and unraveling of processes and mechanisms affecting fruit quality in tomato. The basis of this fruit quality program was a diverse set of 94 cultivated tomato cultivars, representing a wide spectrum of phenotypes for quality related traits. This paper describes a diversity study performed on these cultivars, using information of 882 AFLP markers, of which 304 markers had a known map position. The AFLP markers were scored as much as possible in a co-dominant fashion. We investigated genome distribution and coverage for the mapped markers and conclude that it proved difficult to arrive at a dense and uniformly distributed coverage of the genome with markers. Mapped markers and unmapped markers were used to investigate population structure. A clear substructure was observed which seemed to coincide with a grouping based on fruit size. Finally, we studied amount and decay of linkage disequilibrium (LD) along the chromosomes. LD was observed over considerable (genetic) distances. We discuss the feasibility of marker-trait association studies and conclude that the amount of genetic variation in our set of cultivars is limited, but that there exists scope for association studies. PMID- 18389209 TI - Molecular mapping of an apical branching gene of cultivated sunflower (Helianthus annuus L.). AB - Commercial hybrids of cultivated sunflower (Helianthus annuus L.) are obtained by crossing a cytoplasmic male sterile line (A-line) with a restorer pollinator (R line). The incorporation of a recessive branching trait to extend the pollination period of R-lines during hybrid seed production is laborious and time-consuming. By using target region polymorphism (TRAP) and bulked segregant analysis (BSA), we identified 15 TRAP markers linked to the b(1) (branching) locus in a population of 229 F(2) plants derived from a cross between nonbranched (HA 234) and branched (RHA 271) lines. TBr4-720 and TBr8-555 markers were linked to the b(1) gene in the coupling phase at 0.5 cM (0.004 recombination frequency). The Tbr20-297 and Tbr20-494 markers flanked the b(1) locus in the repulsion phase at genetic distances of 7.5 and 2.5 cM, respectively. Tbr19-395, also in the repulsion phase, mapped at 3.8 cM from the b(1) locus and on the opposite side of the marker Tbr20-297. The 8A1 and 15B3 restriction fragment length polymorphic (RFLP) markers of linkage group (LG) 16 of the RHA 271 x HA 234 cultivated sunflower map anchored the b(1) LG onto the RFLP map. Polymerase chain reaction (PCR)-based markers tightly linked to the recessive b(1) gene have been developed. Their identification and the incorporation of the LG containing the b(1) locus onto an RFLP map will be useful for marker-assisted selection (MAS) in breeding programs and provide the bases for map-based cloning of this gene. PMID- 18389210 TI - Increased osteoclastic activity in acute Charcot's osteoarthropathy: the role of receptor activator of nuclear factor-kappaB ligand. AB - AIMS/HYPOTHESIS: Our aims were to compare osteoclastic activity between patients with acute Charcot's osteoarthropathy and diabetic and healthy controls, and to determine the effect of the receptor activator of nuclear factor-kappaB ligand (RANKL) and its decoy receptor osteoprotegerin (OPG). METHODS: Peripheral blood monocytes isolated from nine diabetic Charcot patients, eight diabetic control and eight healthy control participants were cultured in the presence of macrophage-colony stimulating factor (M-CSF) alone, M-CSF and RANKL, and also M CSF and RANKL with excess concentrations of OPG. Osteoclast formation was assessed by expression of tartrate-resistant acid phosphatase on glass coverslips and resorption on dentine slices. RESULTS: In cultures with M-CSF, there was a significant increase in osteoclast formation in Charcot patients compared with healthy and diabetic control participants (p=0.008). A significant increase in bone resorption was also seen in the former, compared with healthy and diabetic control participants (p<0.0001). The addition of RANKL to the cultures with M-CSF led to marked increase in osteoclastic resorption in Charcot (from 0.264+/-0.06% to 41.6+/-8.1%, p<0.0001) and diabetic control (0.000+/-0.00% to 14.2+/-16.5%, p<0.0001) patients, and also in healthy control participants (0.004+/-0.01% to 10.5+/-1.9%, p<0.0001). Although the addition of OPG to cultures with M-CSF and RANKL led to a marked reduction of resorption in Charcot patients (41.6+/-8.1% to 5.9+/-2.4%, p=0.001), this suppression was not as complete as in diabetic control patients (14.2+/-16.5% to 0.45+/-0.31%, p=0.001) and in healthy control participants (from 10.5+/-1.9% to 0.00+/-0.00%, p<0.0001). CONCLUSIONS/INTERPRETATION: These results indicate that RANKL-mediated osteoclastic resorption occurs in acute Charcot's osteoarthropathy. However, the incomplete inhibition of RANKL after addition of OPG also suggests the existence of a RANKL-independent pathway. PMID- 18389211 TI - Urinary mRNA expression of ACE and ACE2 in human type 2 diabetic nephropathy. AB - AIMS/HYPOTHESIS: The interplay of ACE and type 2 ACE (ACE2) has been recognised as playing an important role in the tissue renin-angiotensin system within the kidney. In the present study, we measured urinary mRNA expression of ACE and ACE2 in patients with type 2 diabetic nephropathy. METHODS: We studied 50 patients with diabetic nephropathy: 26 were being treated by ACE inhibitor (ACEI) alone (ACEI group), the other 24 by ACEI and angiotensin-receptor blocker (ARB) (ACEI+ARB group). mRNA expression of ACE and ACE2 was measured by real-time quantitative RT-PCR at 0 and 12 weeks. All patients were then followed for 56 weeks. RESULTS: Proteinuria correlated significantly with urinary ACE (r=0.454, p=0.001) and ACE2 expression (r=0.651, p<0.001). Urinary ACE2 expression correlated with estimated GFR (r= -0.289, p=0.042). In the ACEI group, there was a significant inverse correlation between the rate of GFR decline and urinary ACE2 expression at baseline (r= -0.423, p=0.031) as well as at 12 weeks (r= 0.395, p=0.046). In contrast, there was no significant correlation between the rate of GFR decline and urinary ACE2 expression at baseline or at 12 weeks in the ACEI+ARB group. The rate of GFR decline did not correlate with the baseline urinary ACE expression of either group. CONCLUSION/INTERPRETATION: There was a relationship between urinary mRNA expression of ACE2 and the degree of proteinuria. The physiological implication and possibility of clinical application of quantifying urinary ACE2 expression require further study. PMID- 18389212 TI - N of 1 trials in diabetes: making individual therapeutic decisions. AB - N of 1 trials are single-subject, randomised, crossover studies, in which the patient serves as their own control to compare the efficacy of a treatment. They offer an alternative to large, randomised clinical trials, which have limited applicability, and empirical testing, which is arbitrary in nature. N of 1 trials are regarded as providing the highest strength of evidence for individual subject decisions. They rely upon the patient expressing preferences and making shared decisions for determining the future therapy, and may therefore lead to the successful implementation of lifestyle interventions. N of 1 trials are an optimal approach for making therapeutic decisions in chronic diseases like diabetes mellitus, where decision-making is often reliant upon arbitrary criteria and clinical judgement. PMID- 18389213 TI - Monitoring of glucose-regulated single insulin secretory granule movement by selective photoactivation. AB - AIMS/HYPOTHESIS: Fluorescence microscopy opens new perspectives for the analysis of insulin secretory granule movement. In this study, we examined whether recently developed photoactivatable/photoconvertible proteins are a useful tool for studying this process at the single granule level in insulin-secreting cells after glucose stimulation. METHODS: Plasmids were generated for expression of fusion proteins of the granule membrane phosphatase phogrin or the granule cargo protein neuropeptide Y (NPY) with the photoactivatable green fluorescent protein mutant A206K (PA-GFP-A206K), the photoconvertible protein Dendra2 and the fluorescent protein mCherry. Transfected insulin-secreting MIN6 cells were analysed by fluorescence microscopy. RESULTS: Point-resolved 405 nm light exposure during image acquisition of MIN6 cells transiently transfected with Phogrin-PA-GFP-A206K or NPY-PA-GFP-A206K as well as of stable MIN6-Phogrin Dendra2 cells resulted in selective visualisation of few granules by green or red fluorescence, respectively. Movement of these granules was analysed by an automated tracking method from confocal 3D image series. The high spatiotemporal resolution facilitated an elongated tracking of single granules. Interestingly, the track speed and track displacement of granules after 1 h starvation and subsequent glucose stimulation was lower in cells pre-cultured for 48 h at 3 mmol/l glucose than in cells pre-cultured at 25 mmol/l glucose. CONCLUSIONS/INTERPRETATION: Targeting of the granule membrane or its cargo with a photoactivatable/photoconvertible protein allows in-depth visualisation and tracking of single insulin granules in dependence upon glucose. This technique may also open the way to elucidating the regulation of granule movement velocity within the pancreatic beta cell with respect to secretory defects in type 2 diabetes. PMID- 18389214 TI - Reconstituted HDL infusion restores endothelial function in patients with type 2 diabetes mellitus. PMID- 18389216 TI - Can transcutaneous oxygen saturation be used to detect tissue hypoperfusion? PMID- 18389215 TI - The effect of different reference transducer positions on intra-abdominal pressure measurement: a multicenter analysis. AB - OBJECTIVE: To investigate the effect of different reference transducer positions on intra-abdominal pressure (IAP) measurement. Three reference levels were studied: the symphysis pubis; the phlebostatic axis; and the midaxillary line at the level of the iliac crest. DESIGN: Prospective cohort study. SETTING: The intensive care units of participating hospitals PATIENTS AND PARTICIPANTS: One hundred thirty-two critically ill patients at risk for intra-abdominal hypertension (IAH). INTERVENTIONS: In each patient, three sets of IAP measurements were obtained in the supine position, using the different reference levels. The IAP measurements obtained at the different reference levels were compared using a paired t-test and Bland-Altman statistics were calculated. MEASUREMENTS AND RESULTS: IAP(phlebostatic) (9.9 +/- 4.67 mmHg) and IAP(pubis) (8.4 +/- 4.60 mmHg) were significantly lower that IAP(midax) (12.2 +/- 4.66 mmHg; p < 0.0001 for both comparisons). The bias between the IAP(midax) and IAP(pubis) was 3.8 mmHg (95% CI 3.5-4.1) and 2.3 mmHg (95% CI 1.9-2.6) between the IAP(midax) and the IAP(phlebostatic). The precision was 3.03 and 3.40, respectively. CONCLUSIONS: In the supine position, IAP(midax) is higher than both IAP(phlebostatic) and IAP(pubis), differences found to be clinically significant; therefore, the symphysis pubis or phlebostatic axis reference lines are not interchangeable with the midaxillary level. PMID- 18389217 TI - Clinical research in critically ill patients: the situation in Italy. PMID- 18389218 TI - Understanding wasted/ineffective efforts in mechanically ventilated COPD patients using the Campbell diagram. PMID- 18389219 TI - Vacuum-assisted closure in the treatment of perineal necrotizing skin and soft tissue infections. PMID- 18389220 TI - Assessment of digoxin antibody use in patients with elevated serum digoxin following chronic or acute exposure. AB - OBJECTIVE: To evaluate the use of antidotal therapy in patients with an elevated digitalis concentration following chronic or acute exposure. DESIGN AND SETTING: Retrospective review of patient records over 2 years in 20 city hospitals in France. PATIENTS: Overall 838 patients with an elevated serum digitalis concentration (digoxin > 1.95ng /ml or digitoxin > 23ng /ml) were included in the study. Of these, 67 (8%) had received antidotal therapy with Fab fragments. MEASUREMENTS AND RESULTS: The relationships between previously reported prognostic criteria and use of antidotal therapy were investigated. We identified five independent factors that were associated with the use of antidotal therapy: acute overdose (OR 15.74), Fab fragment availability in the hospital (11.06), serum potassium (1.81), and heart rate (0.96). Mortality was significantly lower in Fab-treated (6%, 4/67) than untreated patients (15%, 117/770). CONCLUSIONS: Antidotal therapy is underused in patients with an elevated digitalis concentration especially in patients with chronic digitalis exposure. These patients in our series presented a higher mortality rate than patients with acute poisoning. Although they were older and tended to have a history of cardiac disease, they did not differ from patients with acute poisoning with regard to the main severity criteria and prognostic factors. The use of identical criteria for antidotal treatment after acute and chronic poisoning should help optimize outcomes. Fab fragment availability is insufficient in France but ranks only second after type of poisoning (acute or chronic) in the multivariate association with Fab treatment. PMID- 18389221 TI - Computer-assisted glucose control in critically ill patients. AB - OBJECTIVE: Intensive insulin therapy is associated with the risk of hypoglycemia and increased costs of material and personnel. We therefore evaluated the safety and efficiency of a computer-assisted glucose control protocol in a large population of critically ill patients. DESIGN AND SETTING: Observational cohort study in three intensive care units (32 beds) in a 1,300-bed university teaching hospital. PATIENTS: All 2,800 patients admitted to the surgical, neurosurgical, and cardiothoracic units; the study period started at each ICU after implementation of Glucose Regulation for Intensive Care Patients (GRIP), a freely available computer-assisted glucose control protocol. MEASUREMENTS AND RESULTS: We analysed compliance in relation to recommended insulin pump rates and glucose measurement frequency. Patients were on GRIP-ordered pump rates 97% of time. Median measurement time was 5min late (IQR 20min early to 34 min late). Hypoglycemia was uncommon (7% of patients for mild hypoglycemia, < 3.5mmol/l; 0.86% for severe hypoglycemia, < 2.2 mmol/l). Our predefined target range (4.0 7.5 mmol/l) was reached after a median of 5.6 h (IQR 0.2-11.8) and maintained for 89% (70-100%) of the remaining stay at the ICU. The number of measurements needed was 5.9 (4.8-7.3) per patient per day. In-hospital mortality was 10.1%. CONCLUSIONS: Our computer-assisted glucose control protocol provides safe and efficient glucose regulation in routine intensive care practice. A low rate of hypoglycemic episodes was achieved with a considerably lower number of glucose measurements than used in most other schemes. PMID- 18389222 TI - Intracranial self-administration of MDMA into the ventral striatum of the rat: differential roles of the nucleus accumbens shell, core, and olfactory tubercle. AB - RATIONALE: Behavioral and anatomical data suggest that the ventral striatum, consisting of the nucleus accumbens and olfactory tubercle, is functionally heterogeneous. Cocaine and D: -amphetamine appear to be more rewarding when administered into the medial olfactory tubercle or medial accumbens shell than into their lateral counterparts, including the accumbens core. OBJECTIVES: We sought to determine whether rats self-administer the popular recreational drug (+/-)-3,4-methylenedioxymethamphetamine (MDMA) into ventrostriatal subregions and whether the medial olfactory tubercle and medial accumbens shell mediate MDMA's positive reinforcing effects more effectively than their lateral counterparts. RESULTS: Rats receiving 30 mM MDMA into the medial olfactory tubercle, medial accumbens shell, or accumbens core, but not the lateral tubercle or lateral shell, showed higher self-administration rates than rats receiving vehicle. The medial shell supported more vigorous self-administration of MDMA at higher concentrations than the core or medial olfactory tubercle. In addition, intra medial shell MDMA self-administration was disrupted by co-administration of the D1 or D2 receptor antagonists SCH 23390 (1-3 mM) or raclopride (3-10 mM). CONCLUSIONS: Our data suggest that the ventral striatum is functionally heterogeneous. The medial accumbens shell appears to be more important than other ventrostriatal subregions in mediating the positive reinforcing effects of MDMA via both D1- and D2-type receptors. Together with previous data, our data also suggest that unidentified actions of MDMA interfere with the positive reinforcing effects of dopamine in the medial olfactory tubercle. PMID- 18389223 TI - Relationship between wine scores and visible-near-infrared spectra of Australian red wines. AB - Sensory analysis of wine involves the measurement, interpretation and understanding of human responses to the properties perceived by the senses such as sight, smell and taste. The sensory evaluation of wine is often carried out by wine judges, winemakers and technical staff, and allows characterization of the quality of the wine. However, this method is lengthy, expensive, and its results depend on panel training and the specific vocabulary used by the panel. A robust, rapid, unbiased and inexpensive method to assist in quality assessment purposes will therefore be beneficial for the modern wine industry. This study aims to investigate the relationship between sensory analysis, visible (VIS) and near infrared (NIR) spectroscopy to assess sensory properties of commercial Australian wine varieties. For the purposes of this study 118 red wine samples (Cabernet Sauvignon, Shiraz, Pinot Noir, Tempranillo, Nebbiolo and blends) graded by a panel of experienced tasters and scored according to the Australian wine show system were scanned in transmission in the VIS and NIR range (400-2,500 nm). Partial least squares regression models were developed between the overall score given by the judges and the combined VIS-NIR spectra, using full cross validation (leave-one-out method). The results showed that NIR spectroscopy was able to predict wine quality scores in red wine samples (R = 0.61 and standard error of prediction of 0.81). The practical implication of this study is that instrumental methods such as VIS-NIR spectroscopy can be used to complement sensory analysis and can facilitate the task at early stages of product development, making high throughput screening of novel products feasible or maintaining the consistency of the product. PMID- 18389224 TI - An antibody microarray, in multiwell plate format, for multiplex screening of foodborne pathogenic bacteria and biomolecules. AB - Intoxication and infection caused by foodborne pathogens are important problems worldwide, and screening tests for multiple pathogens are needed because foods may be contaminated with multiple pathogens and/or toxic metabolites. We developed a 96-well microplate, multiplex antibody microarray method to simultaneously capture and detect Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium (S. typhimurium), as well as a biomolecule (chicken immunoglobulin G or IgG employed as a proteinaceous toxin analog) in a single sample. Microarrayed spots of capture antibodies against the targeted analytes were printed within individual wells of streptavidin-coated polystyrene 96 multiwell microtiter plates and a sandwich assay with fluorescein- or Cy3-labeled reporter antibodies was used for detection. (Printing was achieved with a conventional microarray printing robot that was operated with custom-developed microplate arraying software.) Detection of the IgG was realized from ca. 5 to 25 ng/mL, and detection of E. coli O157:H7 and S. typhimurium was realized from ca. 10(6) to 10(9) and ca. 10(7) to 10(9) cells/mL, respectively. Multiplex detection of the two bacteria and the IgG in buffer and in culture-enriched ground beef filtrate was established with a total assay (including detection) time of ca. 2.5 h. Detection of S. typhimurium was largely unaffected by high concentrations of the other bacteria and IgG as well as the ground beef filtrate, whereas a small decrease in response was observed for E. coli O157:H7. The multiwell plate, multiplex antibody microarray platform developed here demonstrates a powerful approach for high-throughput screening of large numbers of food samples for multiple pathogens and toxins. PMID- 18389225 TI - NIES certified reference material for microcystins, hepatotoxic cyclic peptide toxins from cyanobacterial blooms in eutrophic water bodies. AB - A certified reference material (CRM) for microcystins has been prepared by the National Institute for Environmental Studies (NIES). Microcystins are hepatotoxic cyclic peptides produced by cyanobacteria in eutrophic water bodies. At least seven microcystin variants were found by HPLC analysis of the NIES CRM, of which [Dha(7)]microcystin-RR and -LR were the major microcystins present. Because of the lack of available standards we determined the total microcystin concentration in the CRM by the MMPB method, and elucidated the structures of the main individual microcystin variants following their isolation. Analyses of NMR and MS spectra indicated that the remaining minor variants in the CRM were [D-Asp(3), Dha(7)]microcystin-RR and -LR, and [Dha(7)]microcystin-YR, -ThTyrR, and -HilR. The CRM is valuable not only as a standard material for the quantitation of total microcystins but also for the identification of individual [Dha(7)]microcystin variants. PMID- 18389226 TI - CE-LIF determination of salivary cadaverine and lysine concentration ratio as an indicator of lysine decarboxylase enzyme activity. AB - Salivary bacteria produce the enzyme lysine decarboxylase which converts lysine to cadaverine. In the absence of appropriate oral hygiene, overgrowth of these bacteria depletes lysine. This may contribute to gingival inflammation, while cadaverine contributes to oral malodor. A selective and sensitive capillary electrophoresis method with laser-induced fluorescence detection has been developed for the determination of cadaverine and lysine in saliva, as an indicator of lysine decarboxylase enzyme activity. The diamino compounds were separated in acidic background electrolyte in their mono-labeled form after derivatization with 4-fluoro-7-nitrobenz-2-oxa-1,3-diazole (NBD-F). Linearity and reproducibility of the method in the range 1-50 MUmol L(-1) have been demonstrated using saliva samples. The method was applied for the measurement of cadaverine and lysine in the saliva of healthy volunteers with or without proper oral hygiene. In the absence of oral hygiene, the mol fraction of cadaverine to cadaverine plus lysine in saliva increased significantly (0.65 +/- 0.13 vs. 0.39 +/- 0.18, P < 0.001), indicating the presence of higher amount of bacterial lysine decarboxylase, that may contribute to periodontal diseases. PMID- 18389227 TI - Detection of low-abundance impurities in synthetic thyroid hormones by stationary phase optimized liquid chromatography-mass spectrometry. AB - The transfer of a gradient method to an isocratic or multistep gradient method employing stationary phase optimized liquid chromatography facilitated a reduction in analysis time by 50% and significantly improved the mass spectrometric detectability of impurities in synthetic thyroid hormones. Four column segments packed with different stationary phases were combined into a single chromatographic column, which allowed the separation and photometric as well as mass spectrometric detection of thyroid compounds in less than 30 min under isocratic- or step gradient elution conditions with 0.10% acetic acid/acetonitrile. Signal instability and baseline drift during detection by negative electrospray ionization time-of-flight mass spectrometry were minimized by optimizing the spray parameters for each individual elution step. This resulted in improved detectabilities and higher mass spectral quality, especially for low-abundance components in the sample mixture. The method was applied to the separation and detection of the low-abundance impurities formed upon the thermal stressing of a sample of synthetic levothyroxine. PMID- 18389228 TI - Cholesterol in women at high cardiovascular risk is less successfully treated than in corresponding men. The Skaraborg Hypertension and Diabetes Project. AB - OBJECTIVE: To assess the use of lipid-lowering therapy in patients with known coronary heart disease (CHD), cerebrovascular disease or diabetes in a community based population in Sweden considering expert recommendations. METHODS: A random sample of individuals aged > or =40 years who were surveyed in 1993-1994 were revisited 10 years later during 2003-2004 (n = 724). A clinical investigation focused on cardiovascular risk including serum total cholesterol. Information on medical history and current medication was collected in structured interviews. RESULTS: Eighty-two patients (11.3%) reported a history of CHD, including 51 men and 31 women. Fifty-three patients fulfilled criteria for treatment and most of them (85%) were on lipid-lowering therapy. A higher fraction of women were treated; however only 13% of them reached target cholesterol levels compared to 37% of the men (P < 0.001). Sixty-five subjects (9.0%) had diabetes and/or a previous stroke (29 men, 36 women) but no previous CHD. Patients with CHD were more likely to be treated compared to patients with diabetes and/or stroke but no CHD (85.0 vs. 28.5%, OR 6.0, 95% CI 2.2-16.9, P = 0.01). In a total of 79 participants (10.9%) who were on lipid-lowering therapy, women reached a total serum cholesterol level below 5.0 mmol/L less often than men (26.3 vs. 63.4%, P < 0.001). CONCLUSIONS: A considerable proportion of patients in primary care were untreated despite current guidelines on lipid-lowering therapy. Treatment outcome in women was less efficient compared with men. Strategies to improve pharmacological treatment in these patients should be developed. PMID- 18389229 TI - Biliary wall calcification in Langerhans cell histiocytosis: report of two cases. AB - Langerhans cell histiocytosis (LCH) is a disorder of unknown pathogenesis affecting one or more organs (unifocal or disseminated form) due to clonal proliferation of Langerhans cells. Liver involvement is more frequent in the disseminated form and the radiological findings of end-stage liver disease due to LCH are similar to those of sclerosing cholangitis. We present the multidetector CT findings in two children with LCH liver involvement and the unique finding of calcification of the biliary wall. PMID- 18389230 TI - Image filtering for two-photon deep imaging of lymphonodes. AB - Non-linear excitation microscopy is considered an ideal spectroscopic method for imaging thick tissues in vivo due to the reduced scattering of infrared radiation. Although imaging has been reported on brain neocortex at 600-800 microm of depth, much less uniform tissues, such as lymphonodes, are characterized by highly anisotropic light scattering that limits the penetration length. We show that the most severe limitation for deep imaging of lymphonodes appears to be the tissue scattering and the diffuse fluorescence emission of labeled cell (lymphocytes) in layers above the focusing plane. We report a study of the penetration depth of the infrared radiation in a model system and in ex vivo lymphonodes and discuss the possibility to apply Fourier filtering to the images in order to improve the observation depth. PMID- 18389231 TI - Spectroscopic and calorimetric studies on the interaction of human serum albumin with DPPC/PEG:2000-DPPE membranes. AB - Site specific spectroscopic techniques and differential scanning calorimetry were used to study human serum albumin (HSA) in the absence and in the presence of membranes composed of dipalmitoylphosphatidylcholine (DPPC) and poly(ethylene glycol:2000)-dipalmitoylphosphatidylethanolamine (PEG:2000-DPPE). Electron spin resonance (ESR) of a maleimide spin-label (5-MSL) covalently bound to the free sulfhydryl group at the unique cystein Cys-34 in domain I, intrinsic fluorescence of the single tryptophan Trp-214 in domain II, and extrinsic fluorescence of p nitrophenyl anthranilate conjugated with tyrosine Tyr-411 in domain III were employed to study HSA dispersions with or without polymer-grafted membranes. On adsorbing at the DPPC membrane surfaces, domain I assumes a more loosened conformation and partitioning of the spin-labelled protein between the aqueous phase and the interfacial region of lipid membranes is observed by ESR. Domain II and III undergo a local structural arrangement which leads Trp-214 and Tyr-411 to come closer and causes intrinsic fluorescence quenching. The influence of DPPC bilayers on HSA is characterized both by a decrease of the thermal unfolding enthalpy and by a slight increase of the transition temperature, Tt, of the protein. The lipid induced effects on HSA are progressively reduced on increasing the amounts of PEG:2000-DPPE mixed with DPPC from the mushroom regime to the brush regime. Primary protein adsorption at the lipid surfaces is abolished at 1 mol% of the polymer-lipid, whereas the secondary protein adsorption at the polymer-brush leads to a further increase of both transition enthalpy and Tt relative to the case of aqueous dispersions of HSA alone. PMID- 18389232 TI - Single locus typing of MHC class I and class II B loci in a population of red jungle fowl. AB - In species with duplicated major histocompatibility complex (MHC) genes, estimates of genetic variation often rely on multilocus measures of diversity. It is possible that such measures might not always detect more detailed patterns of selection at individual loci. Here, we describe a method that allows us to investigate classical MHC diversity in red jungle fowl (Gallus gallus), the wild ancestor of the domestic chicken, using a single locus approach. This is possible due to the well-characterised gene organisation of the 'minimal essential' MHC (BF/BL region) of the domestic chicken, which comprises two differentially expressed duplicated class I (BF) and two class II B (BLB) genes. Using a combination of reference strand-mediated conformation analysis, cloning and sequencing, we identify nine BF and ten BLB alleles in a captive population of jungle fowl. We show that six BF and five BLB alleles are from the more highly expressed locus of each gene, BF2 and BLB2, respectively. An excess of non synonymous substitutions across the jungle fowl BF/BL region suggests that diversifying selection has acted on this population. Importantly, single locus screening reveals that the strength of selection is greatest on the highly expressed BF2 locus. This is the first time that a population of red jungle fowl has been typed at the MHC region, laying the basis for further research into the underlying processes acting to maintain MHC diversity in this and other species. PMID- 18389233 TI - Effect of mixed organic substrate on alpha-tocopherol production by Euglena gracilis in photoheterotrophic culture. AB - Effects of organic carbon sources on cell growth and alpha-tocopherol productivity in wild and chloroplast-deficient W14ZUL strains of Euglena gracilis under photoheterotrophic culture were investigated. In both strains, the increase in cell growth was particularly high when glucose was added as the sole organic carbon source. On the other hand, alpha-tocopherol production per dry cell weight was enhanced by adding ethanol. Ethanol addition also increased the chlorophyll concentration in wild strain and mitochondria activity in W14ZUL strain. For effective alpha-tocopherol production, the effects of mixture of glucose and ethanol were investigated. The results showed that, when a mixture of glucose (6 g/l) and ethanol (4 g/l) was used, alpha-tocopherol productivity per culture broth was 3.89 x 10(-2) mg l(-1) h(-1), which was higher than the value obtained without addition of organic carbon source (0.92 x 10(-2) mg l(-1) h(-1)). In addition, under fed-batch cultivation using an internally illuminated photobioreactor, the alpha-tocopherol production per culture broth was 23.43 mg/l, giving a productivity of 16.27 x 10(-2) mg l(-1) h(-1). PMID- 18389234 TI - The tryptophan synthetase gene TRP1 of Nodulisporium sp.: molecular characterization and its relation to nodulisporic acid A production. AB - Nodulisporic acid A (NAA), an insecticidal indole diterpene, is produced by the fungus Nodulisporium sp. Since indole-3-glycerolphosphate is the precursor of the indole moiety of NAA, it is suggested that the activity of tryptophan synthetase may play a role in NAA biosynthesis. To investigate this hypothesis, the tryptophan synthetase gene TRP1 of Nodulisporium sp. was cloned and characterized. The gene consists of three introns of 146, 68, and 57 bp. The four exons encode a protein of 712 amino acids, the sequence of which is highly homologous to that of other fungal tryptophan synthetase proteins. The transcription initiation site was mapped 66 bp upstream to the ATG, and the polyA tail attachment site is 169 bp downstream to the translation stop codon. Replacement of the N-terminal half of the gene with a hygromycin selection marker yielded mutants with the tryptophan auxotroph/hygromycin-resistance (trp(-)/hyr) phenotype. The TRP1 mutants required a high concentration of tryptophan supplement in solid medium (10 mM) to sustain minimal growth and failed to produce NAA in the production medium (FFL-CAM) supplemented with high concentrations of tryptophan. PMID- 18389235 TI - Detection of recombinant Pseudomonas putida in the wheat rhizosphere by fluorescence in situ hybridization targeting mRNA and rRNA. AB - A method was developed to detect a specific strain of bacteria in wheat root rhizoplane using fluorescence in situ hybridization and confocal microscopy. Probes targeting both 23S rRNA and messenger RNA were used simultaneously to achieve detection of recombinant Pseudomonas putida (TOM20) expressing toluene o monooxygenase (tom) genes and synthetic phytochelatin (EC20). The probe specific to P. putida 23S rRNA sequences was labeled with Cy3 fluor, and the probe specific to the tom genes was labeled with Alexa647 fluor. Probe specificity was first determined, and hybridization temperature was optimized using three rhizosphere bacteria pure cultures as controls, along with the P. putida TOM20 strain. The probes were highly specific to the respective targets, with minimal non-specific binding. The recombinant strain was inoculated into wheat seedling rhizosphere. Colonization of P. putida TOM20 was confirmed by extraction of root biofilm and growth of colonies on selective agar medium. Confocal microscopy of hybridized root biofilm detected P. putida TOM20 cells emitting both Cy3 and Alexa647 fluorescence signals. PMID- 18389236 TI - [Benign epithelial odontogenic tumors]. AB - The group of benign epithelial odontogenic tumors consists of the four member types of the ameloblastoma family (solid/multicystic, extraosseous/peripheral, desmoplastic, unicystic), squamous odontogenic tumors, calcifying odontogenic tumors, adenomatoid odontogenic tumors, and keratocystic odontogenic tumors, the former "keratocysts" that were recently reclassified by the World Health Organization and are now regarded as tumors. The latter are by far the most frequent tumors in this group, followed by solid/multicystic ameloblastoma. Although the etiology of these lesions is still unknown, a close relationship to normal tooth development is obvious, which is partially imitated by some tumors. Despite some similarities to each other, at least in part, the biological behavior of these lesions is quite different, as are treatment modalities. The diagnosis is essentially based on localization (intraosseous vs. extraosseous/peripheral) and histology, whereupon the correlation of histological findings with radiographic morphology may be of additional diagnostic value. Because of the range of variation, immunohistochemical investigations are not helpful in diagnosing a particular case. PMID- 18389237 TI - [Thymic cyst with initial malignant transformation]. AB - We present the case of a 38-year-old woman with a thymus cyst with an incidental tiny adenocarcinoma. Adenocarcinomas and adenosquamous carcinomas of the thymus or in thymus cysts have seldom been reported. These cases were overt invasive carcinomas. Our case is the first report on an incipient adenocarcinoma in a thymus cyst. It demonstrates the importance of thorough histological examination of cystic changes in the thymus, not only for the diagnostic classification of the lesion but also to ensure that a possible malignant change is not overlooked. PMID- 18389238 TI - Associated thyreoiditis, myasthenia gravis, thymectomy, Chron's disease, and erythema nodosum: pathogenetic and clinical correlations, immune system involvement, and systemic infectious complications. AB - The case of a young woman suffering from multiple autoimmune-dysreactive disorders (including thyreoiditis, myasthenia gravis, thymectomy, Crohn's disease, and erythema nodosum), while undergoing steroideal therapy, was complicated by a severe infectious disorder (severe upper urinary tract infection). While the pathogenetic and clinical relationship between the different autoimmune-dysreactive complications is still unclear, and the supporting role of the frequent immunosuppressive treatment may add significantly to these risk factors, clinicians who are engaged in the management of these patients should be aware that multiple, concurrent or subsequent disorders might occur in these subjects, and also that severe infections might be of relevant concern. PMID- 18389239 TI - Massive gastrointestinal bleeding due to the rupture of arterial aneurysm in Behcet's disease: case report and literature review. AB - Massive gastrointestinal bleeding is a very rare manifestation of gastrointestinal Behcet's disease, mainly from the gastrointestinal mucosal lesions. We report herein the case of a 50-year-old man with intestinal Behcet's disease who suffered massive hemorrhage from ruptured arterial aneurysm. Colonoscopy demonstrated large amount of fresh blood in the entire colon, but we were not able to localize bleeding focus anywhere in the colon. Angiography was performed and it revealed that a small aneurysm on the right ileocolic artery with apparent extravasation of contrast material. A guiding catheter was inserted to a right ileocolic artery and superselective arterial embolization using microcoils was successful. Following this procedure, the gastrointestinal bleeding gradually subsided and completely stopped within a few days. He is now treating with prednisolone and sulfasalazine without recurrent bleeding until now. PMID- 18389240 TI - Effects of pulsed and sinusoid electromagnetic fields on human chondrocytes cultivated in a collagen matrix. AB - Although several effects of electromagnetic fields (EMFs) on articular cartilage have been reported in recent studies, the use of EMFs to treat osteoarthritis remains a matter of debate. In an in vitro study, human chondrocytes harvested from osteoarthritic knee joints were released from their surrounding matrix and transferred in defined concentration into a 3D matrix (type-I collagen gel). The cultivation, performed under standard conditions, lasted up to 14 days. During this time, treatment groups were continuously exposed to either sinusoid or pulsed electromagnetic fields (PEMFs). The PEMFs revealed the following characteristics: maximum magnetic flux density of 2 mT, frequency of the bursts of 16.7 Hz with each burst consisting of 20 pulses. Similarly, the sinusoid EMFs also induced a maximum flux density of 2 mT with a frequency of 50 Hz. Control groups consisting of equal number of samples were not exposed to EMF. Immunohistological examinations of formalin-fixed, paraffin-embedded samples revealed positive staining for type-II collagen and proteoglycans in the immediate pericellular region with no differences between the two different treatment groups and the control groups. With increasing cultivation time, both type-II collagen and aggrecan gene expression declined, but no significant differences in gene expression were found between the treatment and control groups. In conclusion, using our in vitro setting, we were unable to detect any effects of pulsed and sinusoidal magnetic fields on human adult osteoarthritic chondrocytes. PMID- 18389241 TI - Iodine contrast media doses equal-attenuating with gadolinium chelates at CT aortography may have less risk of contrast-induced nephropathy and no risk of nephrogenic systemic fibrosis in azotaemic patients! PMID- 18389242 TI - CT radiation dose in children: a survey to establish age-based diagnostic reference levels in Switzerland. AB - This work aimed at assessing the doses delivered in Switzerland to paediatric patients during computed tomography (CT) examinations of the brain, chest and abdomen, and at establishing diagnostic reference levels (DRLs) for various age groups. Forms were sent to the ten centres performing CT on children, addressing the demographics, the indication and the scanning parameters: number of series, kilovoltage, tube current, rotation time, reconstruction slice thickness and pitch, volume CT dose index (CTDI(vol)) and dose length product (DLP). Per age group, the proposed DRLs for brain, chest and abdomen are, respectively, in terms of CTDI(vol): 20, 30, 40, 60 mGy; 5, 8, 10, 12 mGy; 7, 9, 13, 16 mGy; and in terms of DLP: 270, 420, 560, 1,000 mGy cm; 110, 200, 220, 460 mGy cm; 130, 300, 380, 500 mGy cm. An optimisation process should be initiated to reduce the spread in dose recorded in this study. A major element of this process should be the use of DRLs. PMID- 18389243 TI - Bone marrow MR imaging as predictors of outcome in hemopoietic stem cell transplantation. AB - The purpose of this study is to investigate the role of femoral marrow MR imaging as predictor of outcome for hemopoietic stem cell transplantation (HSCT) in beta thalassemia major. MR imaging of the proximal femur, including T1- and T2 weighted spin echo and short-tau inversion recovery and in-phase and out-of-phase fast field echo images, was prospectively performed in 27 thalassemia major patients being prepared for HSCT. The area of red marrow and its percentage of the proximal femur were measured, and the presence of marrow hemosiderosis was assessed. Age-adjusted multivariate logistic regression was used to determine the relationship between red marrow area percentage and marrow hemosiderosis and HSCT outcome. Red area percentage were less in patients with successful (90.25 +/- 4.14%) compared to unsuccessful transplants (94.54% +/- 2.93%; p = 0.01). Red marrow area percentage correlated positively with duration of symptoms(r = 0.428, p = 0.026) and serum ferritin (r = 0.511, p = 0.006). In multivariate-adjusted logistic regression analyses, red marrow area percentage was significantly inversely associated with successful HSCT (OR = 1.383, 95% CI: 1.059-1.805, p = 0.005). Marrow hemosidersosis and duration of sympotms and serum ferritin were not associated with HSCT outcome(p = 0.174, 0.974, 0.762, respectively). Red marrow area percentage of proximal femur on MR imaging is a useful predictor of HSCT outcome. PMID- 18389244 TI - CT assessment of herniation pits: prevalence, characteristics, and potential association with morphological predictors of femoroacetabular impingement. AB - The purpose of this study was to evaluate whether there is a correlation between the presence of herniation pits (HPs) and morphological indicators of cam and pincer femoroacetabular impingement (FAI) based on computed tomography (CT) examinations. CT studies of the pelvis obtained from 200 patients were retrospectively analysed for the presence of HPs and morphological abnormalities of the femoral head and acetabulum. As an indicator for cam FAI, we used the angle alpha, describing the anterior femoral head-neck junction. As an indicator for pincer FAI, we measured the acetabular coverage and the acetabular orientation. Student's t-test was used for statistical analysis. HPs were identified in 85 of the 200 patients. HPs were predominantly found in the superior portion of the proximal anterior femoral neck; some were located in the inferior portion. The angle alpha was significantly larger by 10% in the group with HPs. A correlation between the presence of HPs and morphological indicators of pincer FAI was not found. In conclusion HPs are not only located in the superior portion of the proximal anterior femoral neck, but also in the inferior portion. There is an association between the presence of HPs and a high value of angle alpha. PMID- 18389245 TI - Pelvic floor muscle lesions at endoanal MR imaging in female patients with faecal incontinence. AB - To evaluate the frequency and spectrum of lesions of different pelvic floor muscles at endoanal MRI in women with severe faecal incontinence and to study their relation with incontinence severity and manometric findings. In 105 women MRI examinations were evaluated for internal anal sphincter (IAS), external anal sphincter (EAS), puborectal muscle (PM) and levator ani (LA) lesions. The relative contribution of lesions to differences in incontinence severity and manometric findings was studied. IAS (n = 59) and EAS (n = 61) defects were more common than PM (n = 23) and LA (n = 26) defects. PM and LA defects presented mainly with IAS and/or EAS defects (isolated n = 2 and n = 3). EAS atrophy (n = 73) was more common than IAS (n = 19), PM (n = 16) and LA (n = 9) atrophy and presented mainly isolated. PM and LA atrophy presented primarily with EAS atrophy (isolated n = 3 and n = 1). Patients with IAS and EAS lesions had a lower resting and squeeze pressure, respectively; no other associations were found. PM and LA lesions are relatively common in patients with severe faecal incontinence, but the majority of lesions are found in women who also have IAS and/or EAS lesions. Only an association between anal sphincter lesions and manometry was observed. PMID- 18389246 TI - Proton MR spectroscopy of cerebral gliomas at 3 T: spatial heterogeneity, and tumour grade and extent. AB - This study aimed to evaluate the usefulness of proton MR spectroscopic imaging ((1)H-MRSI) at 3 T in differentiating high- from low-grade gliomas, and tumour from necrosis, oedema or normal tissue. Forty-four patients with brain gliomas and four with meningiomas were retrospectively reviewed. The normalised metabolites choline (nCho), N-acetylaspartate (nNAA), creatine (nCr) and lactate/lipids (nLL), and the metabolite ratios Cho/NAA, NAA/Cr and Cho/Cr were calculated. Necrotic-appearing areas showed two spectroscopic patterns: "necrosis" with variable nCho and high nLL, and "cystic necrosis" with variable nLL or nonevident peaks. Peri-enhancing oedematous-appearing areas showed three spectroscopic patterns ("tumour" with abnormal Cho/NAA, "oedema" with normal Cho/NAA and "tumour/oedema" with normal nCho and abnormal Cho/NAA) in gliomas, and one ("oedema") in meningiomas. Peri-enhancing or peri-tumour normal-appearing areas showed two patterns ("infiltrated" with abnormal nCho and/or Cho/NAA and "normal" with normal spectra) in gliomas and one ("normal") in meningiomas. Discriminant analysis showed that classification accuracy between high- and low grade glioma masses was better with normalised metabolites or all parameters together than metabolite ratios and that among peri-enhancing areas was much better with normalised metabolites. The analysis of spatial distribution of normalised metabolites by 3-T (1)H-MRSI helps to discriminate among different tissues, offering information not available with conventional MRI. PMID- 18389247 TI - Cranial CT with 64-, 16-, 4- and single-slice CT systems-comparison of image quality and posterior fossa artifacts in routine brain imaging with standard protocols. AB - Posterior fossa artifacts constitute a characteristic limitation of cranial CT. To identify practical benefits and drawbacks of newer CT systems with reduced collimation in routine cranial imaging, we aimed to investigate image quality, posterior fossa artifacts and parenchymal delineation in non-enhanced CT (NECT) with 1-, 4-, 16- and 64-slice scanners using standard scan protocols. We prospectively enrolled 25 consecutive patients undergoing NECT on a 64-slice CT. Three groups with 25 patients having undergone NECT on 1-, 4- and 16-slice CT machines were matched regarding age and sex. Standard routine CT parameters were used on each CT system with helical acquisition in the posterior fossa; the parameters varied regarding collimation and radiation dose. Three blinded readers independently assessed the cases regarding image quality, infra- and supratentorial artifacts and delineation of brain parenchymal structures on a five-point ordinal scale. Reading orders were randomized. A proportional odds model that accounted for the correlated nature of the data was fit using generalized estimating equations. Posterior fossa artifacts were significantly reduced, and the delineation of infratentorial brain structures was significantly improved with the thinner collimation used for the newer CT systems (p<0.001). No significant differences were observed for midbrain structures (p>0.5). The thinner collimation available on modern CT systems leads to reduced posterior fossa artifacts and to a better delineation of brain parenchyma in the posterior fossa. PMID- 18389248 TI - CT colonography: computer-aided detection of morphologically flat T1 colonic carcinoma. AB - The purpose was to evaluate the ability of computer-aided detection (CAD) software to detect morphologically flat early colonic carcinoma using CT colonography (CTC). Twenty-four stage T1 colonic carcinomas endoscopically classified as flat (width over twice height) were accrued from patients undergoing staging CTC. Tumor location was annotated by three experienced radiologists in consensus aided by the endosocpic report. CAD software was then applied at three settings of sphericity (0, 0.75, and 1). Computer prompts were categorized as either true positive (overlapping tumour boundary) or false positive. True positives were subclassified as focal or non focal. The 24 cancers were endoscopically classified as type IIa (n=11) and type IIa+IIc (n=13). Mean size (range) was 27 mm (7-70 mm). CAD detected 20 (83.3%), 17 (70.8%), and 13 (54.1%) of the 24 cancers at filter settings of 0, 0.75, and 1, respectively with 3, 4, and 8 missed cancers of type IIa, respectively. The mean total number of false-positive CAD marks per patient at each filter setting was 36.5, 21.1, and 9.5, respectively, excluding polyps. At all settings, >96.1% of CAD true positives were classified as focal. CAD may be effective for the detection of morphologically flat cancer, although minimally raised laterally spreading tumors remain problematic. PMID- 18389249 TI - Endovascular coils as lung tumour markers in real-time tumour tracking stereotactic radiotherapy: preliminary results. AB - To evaluate the use of endovascular coils as markers for respiratory motion correction during high-dose stereotactic radiotherapy with the CyberKnife, an image-guided linear accelerator mounted on a robotic arm. Endovascular platinum embolisation coils were used to mark intrapulmonary lesions. The coils were placed in subsegmental pulmonary artery branches in close proximity to the target tumour. This procedure was attempted in 25 patients who were considered unsuitable candidates for standard transthoracic percutaneous insertion. Vascular coils (n = 87) were successfully inserted in 23 of 25 patients. Only minor complications were observed: haemoptysis during the procedure (one patient), development of pleural pain and fever on the day of procedure (one patient), and development of small infiltrative changes distal to the vascular coil (five patients). Fifty-seven coils (66% of total inserted number) could be used as tumour markers for delivery of biologically highly effective radiation doses with automated tracking during CyberKnife radiotherapy. Endovascular markers are safe and allow high-dose radiotherapy of lung tumours with CyberKnife, also in patients who are unsuitable candidates for standard transthoracic percutaneous marker insertion. PMID- 18389250 TI - Reproducibility over a 1-month period of 1H-MR spectroscopic imaging NAA/Cr ratios in clinically stable multiple sclerosis patients. AB - N-acetylaspartate/creatine (NAA/Cr) ratios, assessed with proton magnetic resonance spectroscopy, are increasingly used as a surrogate marker for axonal dysfunction and degeneration in multiple sclerosis (MS). The purpose of this study was to test short-time reproducibility of NAA/Cr ratios in patients with clinically stable MS. In 35 MS patients we analysed NAA/Cr ratios obtained with (1)H-MR spectroscopic imaging at the centrum semiovale either with lateral ventricles partially included (group 1; n=15) or more cranially with no ventricles included (group 2; n=20). To test short-term reproducibility of the NAA/Cr measurements, patients were scanned twice 4 weeks apart. We determined mean NAA/Cr and Cho/Cr ratios of 12 grey matter and 24 white matter voxels. Mean NAA/Cr ratios of both the white and grey matter did not change after 4 weeks. Overall 4-week reproducibility of the NAA/Cr ratio, expressed as coefficient of variation, was 4.8% for grey matter and 3.5% for white matter. Reproducibility of cranial scanning of the ventricles was slightly better than with cerebrospinal fluid included. Our study shows good short-term reproducibility of NAA/Cr ratio measurements in the centrum semiovale, which supports the reliability of this technique for longitudinal studies. PMID- 18389251 TI - Neuroradiologic findings in children with mitochondrial disorder: correlation with mitochondrial respiratory chain defects. AB - Mitochondrial disorders are a heterogeneous group of disorders affecting energy metabolism that can present at any age with a wide variety of clinical symptoms. We investigated brain magnetic resonance (MR) findings in 40 children with defects of the mitochondrial respiratory chain (MRC) complex and correlated them with the type of MRC defects. Enrolled were 40 children with MRC defects in biochemical enzyme assay of the muscle specimen. Twenty-one children were found to have classical syndromes of mitochondrial disorders and 19 children presented nonspecific mitochondrial encephalomyopathies. Their brain MR imaging findings were retrospectively reviewed and correlated with the biochemical defect in the MRC complex. Children with MRC defects showed various neuroradiologic features on brain MR imaging that resulted from a complex genetic background and a heterogeneous phenotype. Rapid progression of atrophy involving all structures of the brain with variable involvement of deep gray and white matter are the most frequent MR findings in children with MRC defects in both classical syndromes of mitochondrial disorder and nonspecific mitochondrial encephalomyopathies. The type of biochemical defect in the MRC complex enzyme did not correlate with brain MR findings in child patients. PMID- 18389252 TI - Percutaneous radiofrequency ablation of hepatocellular carcinoma: analysis of 80 patients treated with two consecutive sessions. AB - This study investigated the reasons for some patients requiring two consecutive sessions of percutaneous radiofrequency (RF) ablation of hepatocellular carcinoma (HCC). We reviewed our database of 1,179 patients (1,624 treatments) with HCCs treated by percutaneous ultrasound (US)-guided RF ablation over 6 years. We retrospectively evaluated 80 patients who required a second session after the first session. The medical records and follow-up CTs were studied. We assessed the reasons for the second session and the patient outcomes. A second session was required in 80 (4.8%) out of 1,642 treatments of percutaneous RF ablation for HCC. The reason for the second session included technical failure related to the patient or the procedure (n=26), technical failure due to residual (n=40), newly detected (n=11) or missed (n=3) tumors found at the immediate follow-up CT. All patients were retreated with a second RFA session the next day. Seventy-five (93%) of 80 patients achieved complete ablation after the second session. The remaining five patients were treated by TACE (n=1), additional RFA (as second treatment at next admission) (n=3), or were lost to follow-up (n=1). After 1 month follow-up, 72 patients (96%) showed complete ablation after the second session. The interventional oncologist should understand the technical reasons for a patient requiring a second session of RF ablation when providing treatment for HCCs and perform careful pre-procedural planning to minimize the need for multi-session procedures. PMID- 18389254 TI - Renal cell carcinoma. PMID- 18389253 TI - Breast MRI: guidelines from the European Society of Breast Imaging. PMID- 18389255 TI - Neuronal correlates of the visually elicited escape response of the crab Chasmagnathus upon seasonal variations, stimuli changes and perceptual alterations. AB - When confronted with predators, animals are forced to take crucial decisions such as the timing and manner of escape. In the case of the crab Chasmagnathus, cumulative evidence suggests that the escape response to a visual danger stimulus (VDS) can be accounted for by the response of a group of lobula giant (LG) neurons. To further investigate this hypothesis, we examined the relationship between behavioral and neuronal activities within a variety of experimental conditions that affected the level of escape. The intensity of the escape response to VDS was influenced by seasonal variations, changes in stimulus features, and whether the crab perceived stimuli monocularly or binocularly. These experimental conditions consistently affected the response of LG neurons in a way that closely matched the effects observed at the behavioral level. In other words, the intensity of the stimulus-elicited spike activity of LG neurons faithfully reflected the intensity of the escape response. These results support the idea that the LG neurons from the lobula of crabs are deeply involved in the decision for escaping from VDS. PMID- 18389256 TI - Representation of a mixture of pheromone and host plant odor by antennal lobe projection neurons of the silkmoth Bombyx mori. AB - Pheromone-source orientation behavior can be modified by coexisting plant volatiles. Some host plant volatiles enhance the pheromonal responses of olfactory receptor neurons and increase the sensitivity of orientation behavior in the Lepidoptera species. Although many electrophysiological studies have focused on the pheromonal response of olfactory interneurons, the response to the mixture of pheromone and plant odor is not yet known. Using the silkmoth, Bombyx mori, we investigated the physiology of interneurons in the antennal lobe (AL), the primary olfactory center in the insect brain, in response to a mixture of the primary pheromone component bombykol and cis-3-hexen-1-ol, a mulberry leaf volatile. Application of the mixture enhanced the pheromonal responses of projection neurons innervating the macroglomerular complex in the AL. In contrast, the mixture of pheromone and cis-3-hexen-1-ol had little influence on the responses of projection neurons innervating the ordinary glomeruli whereas other plant odors dynamically modified the response. Together this suggests moths can process plant odor information under conditions of simultaneous exposure to sex pheromone. PMID- 18389258 TI - Depressed skull fracture and epidural hematoma from head fixation with pins for craniotomy in children. AB - OBJECTIVE: A head fixation device with pins is commonly used for immobilization of the patient's head during craniotomy. The safety of head fixation devices in children has been discussed rarely in the literature. The purpose of this report is to review our experience with complications of head fixation with pins in children undergoing craniotomies and to review the literature on this subject. MATERIALS AND METHODS: The database of the Division of Neurosurgery was reviewed to identify children who had cranial complications related to the use of a pin head fixation device. The charts of these patients were reviewed retrospectively. RESULTS: Five of 766 children (0.65%) undergoing craniotomies with pin fixation of the head had depressed skull fractures and/or epidural hematomas from the pin fixation. Age ranged from 2.6 to 7.5 years; all fractures were temporal and occurred during posterior fossa craniotomies. CONCLUSIONS: Depressed skull fractures and associated epidural hematomas need to be considered as possible complications of pin fixation of the head for craniotomy in young children. PMID- 18389259 TI - Endoscopic treatment of suprasellar arachnoid cysts. AB - OBJECTIVE: Arachnoid cysts constitute 1% of all intracranial mass lesions not resulting from trauma. Suprasellar arachnoid cysts (SACs) are uncommon. Obstructive hydrocephalus is the most common cause of initial symptoms and occurs in almost 90% of the patients with suprasellar arachnoid cyst. We report on 17 patients with suprasellar arachnoid cyst who were treated with neuroendoscopic intervention. MATERIALS AND METHODS: Seventeen patients with SAC had been operated on between 1999 and 2007 in our institution. Five patients had previously undergone shunting procedures or craniotomy. Nine boys and eight girls ranged in age from 4 months to 17 years (mean 53.2 months). All patients had hydrocephalus. A wide ventriculocystostomy (VC) or a ventriculo-cysto cisternostomy (VCC) was done by using rigid neuroendoscopes. Psychometric evaluation was administered postoperatively, when possible. Follow-up of the patients ranged from 6 months to 7 years (mean 41.6 months). RESULTS: Of the 17 patients, 12 underwent endoscopic procedure as the primary surgery. VC only was performed in the first patient. In the remaining 16 patients, VCC was done. A slit-valve mechanism was observed in 14 patients. Three patients needed a VP shunt, despite a successful VCC. Three of five patients, younger than 1 year of age became shunt dependent and none of the patients older than 1 year of age needed shunting. In four patients presented with shunt malfunction, the shunts were removed after endoscopic surgery. Only one subject's total score was under the normal IQ limits but her subtests scores were heterogeneous. However, there was marked discrepancy among her subtests scores. CONCLUSION: Endoscopic surgery should be the first choice in the management of SACs. Neuroendoscopic VCC is successful in the majority of the cases. PMID- 18389260 TI - Fusions at the craniovertebral junction. AB - INTRODUCTION: The surgical management of craniovertebral junction instability in pediatric patients has unique challenges. While the indications for internal fixation in children are similar to those of adults, the data concerning techniques, complications, and outcomes of spinal instrumentation comes from experience with adult patients. Diminutive osseous and ligamentous structures and anatomical variations associated with syndromic craniovertebral abnormalities frequently complicates the approaches and limits the use of internal fixation in children. Cervical arthrodesis in the pediatric age group has the potential for limiting growth potential and causing secondary deformity. Recent advances in image analysis have enabled preoperative planning which is critical to evaluate the size of instrumentation and its relation to the patient's anatomy. Newer techniques have recently evolved and have been incorporated in the management of pediatric patients with requirement for craniocervical stabilization. MATERIALS AND METHODS: Over 750 craniovertebral junction fusions have been reviewed in children. The indications for atlantoaxial arthrodesis were: (a) absent odontoid process, dystopic os odontoideum, absent posterior arch of C1; (b) Morquio's syndrome, Goldenhar's syndrome, Conradi's syndrome, and spondyloepiphyseal dysplasia. The acquired abnormalities of trauma, postinfectious instability, and Down's syndrome completed the indication in children. The indications for occipitocervical fusion were: (a) anterior and posterior bifid C1 arches with instability, absent occipital condyles; b) severe reducible basilar invagination, unstable dystopic os odontoideum, and unilateral atlas assimilation; (c) acquired phenomenon with traumatic occipitocervical dislocation, complex craniovertebral junction fractures of C1 and C2, after transoral craniovertebral junction decompression, cranial settling in Down's syndrome and inflammatory disease such as Grisel's syndrome. Instability was seen in children with clivus chordoma and osteoblastoma. Atlantoaxial fusions were performed mainly with interlaminar rib graft fusion and more recently with the transarticular screw fixation in the older patient. In the teenager, lateral mass screws at C1 and rod fixation were made; C2 pars interarticular screw fixation and C2 pedicle screw fixation. A C2 translaminar screw fixation is described. Occipitocervical fusions were made utilizing rib grafts below the age of 6. A contoured loop fixation was made in children above the age of 7, and recently, rod and screw fixation was also utilized. RESULTS: Abnormal cervical spine growth was not seen in children who underwent craniocervical stabilization below the age of 5. The authors have reserved rigid instrumentation for children above the age of 10 years and dependent on the anatomy. PMID- 18389261 TI - Anatomy and biomechanics of normal craniovertebral junction (a) and biomechanics of stabilization (b). AB - INTRODUCTION: A knowledge of the bony configuration, ligamentous attachments, joint articulations, vascular supply, muscle function, and lymphatic drainage as well as the kinetic anatomy of the craniocervical junction is necessary to understand the etiology of abnormalities in this area and their treatment. RESULTS AND DISCUSSION: The craniovertebral junction (CVJ) is the most mobile of the upper cervical spine especially in children. It is uniquely adapted for stability and motion. The bony anatomy and the normal biomechanics of the CVJ in children are presented and subsequently the biomechanics of complex stabilization. Our review of more than 600 children who required stabilization is presented. PMID- 18389262 TI - Surgical approaches: postoperative care and complications "transoral transpalatopharyngeal approach to the craniocervical junction". AB - INTRODUCTION: The ventral approach to the craniocervical border has been described for decompression of irreducible extradural pathology. The procedures utilized encompass the transoropharyngeal and median mandibulotomy with glossotomy and the transpalatal procedures. This study was aimed to review the utility of the transoral-transpalatopharyngeal approach. CLINICAL MATERIALS AND METHODS: Seven hundred thirty-three patients underwent transpalatopharyngeal approach for decompression of the brain stem and cervicomedullary junction. Of these, 280 were children below the age of 16 years. The main indication was irreducible ventral pathology compressing the brain stem and cervicomedullary junction. Two hundred two children had irreducible basilar invagination, 28 had proatlas segmentation abnormalities, os odontoideum with a dystopic os odontoideum in 30, and spinal tumors in seven (chordoma, fibrous dysplasia, osteoblastoma). Seven patients with Down's syndrome and irreducible bony compression of the ventral cervicomedullary junction were seen. There were six other miscellaneous diagnoses. All children required craniocervical stabilization which was carried out under the same anesthetic as the transoral procedure. OPERATIVE PROCEDURE: The procedure entailed fiber-optic intubation. The patient was placed in cervical traction prior to the anterior procedure. The soft palate was split only in individuals with a short clivus with a high riding clivus odontoid articulation. Craniocervical stabilization was performed in the prone position under the same anesthetic. RESULTS: There was one retropharyngeal infection postoperatively. No cesium fluoride leaks were encountered. Velopalatine incompetence was seen in five children who already had preoperative brain stem dysfunction. Neurological recovery was the rule. Patients who had preoperative syringohydromyelia had resolution of the syrinx on postoperative magnetic resonance imaging. DISCUSSION: The author's technique is described. Since 1977, the procedure has been performed in 732 patients (280 children) and has evolved into a safe and direct approach to the ventral cervicomedullary junction with minimal morbidity and mortality. PMID- 18389264 TI - Short-term results of hip revisions with a curved cementless modular stem in association with the surgical approach. AB - INTRODUCTION: A prospective study was designed to test the hypothesis that short term results after hip revisions are in association with the surgical approach with lower clinical scores for the transfemoral approach. MATERIALS AND METHODS: A total of 120 hip revision operations with the modular cementless revision stem "Revitan Curved" (Zimmer GmbH, Winterthur, Switzerland), of which 42 replacements involved an endofemoral (posterolateral) approach and 78 implantations a transfemoral approach, were followed up over a period of at least 24 months. RESULTS: In the early post-operative stage, stems implanted transfemoral were associated with significantly lower Harris Hip Scores and a significantly more frequent appearance of Trendelenburg signs. The differences lessened at the end of the follow-up period. Within the group of transfemoral implantation, all six stems with a circular fixation zone measuring less than 3 cm had subsided and two of these had become loose; none of the stems with greater fixation zones exhibited these properties. In the case of the endofemoral implants, three stems exhibited sinking but there did not appear to be any relationship between this event and length of fixation zone. CONCLUSION: The surgical approach has an association with the short-time outcomes of hip revsions using cementless modular stems. Transfemoral implantation of the "Revitan curved" stem requires a fixation zone of at least 3 cm and a longer period of rehabilitation. PMID- 18389263 TI - Clinical, neuropathological and genotypic variability in SNCA A53T familial Parkinson's disease. Variability in familial Parkinson's disease. AB - Individuals with familial Parkinson's disease (PD) due to a monogenic defect can show considerable clinical and neuropathological variability. To identify factors underlying this variability, histopathological analysis was performed in two clinically different A53T alpha-synuclein heterozygotes from Family H, a multigenerational alpha-synuclein A53T kindred. To determine whether additional genetic factors could contribute to phenotypic variability, Family H and another multigenerational A53T kindred were analyzed for parkin polymorphisms. We identified a previously described variant in parkin exon 4 associated with increased PD risk (S167N). The two A53T heterozygotes had markedly different neuropathology and different parkin genotypes: A N167 homozygote had early onset rapidly progressive disease, early dementia, myoclonus and sleep disorder, while a S167 homozygote had late onset, slowly progressive disease and late dementia. Both had brainstem, cortical, and intraneuritic Lewy bodies (LB). The N167 individual had widespread cortical neurofibrillary degeneration, while the S167 individual had only medial temporal lobe neurofibrillary degeneration. The N167 individual had severe neuronal loss in CA2 associated with Lewy neurites (LN), while the S167 individual had severe neuronal loss in CA1 associated with TDP-43 immunoreactive neuronal inclusions. These findings implicate TDP-43 in the pathology of familial PD and suggest that parkin may act as a modifier of the A53T alpha-synuclein phenotype of familial PD. Furthermore, they suggest a mechanism by which a rare genetic variant that is associated with a minor increase of PD risk in the heterozygous state may, in the homozygous state, exacerbate a disease phenotype associated with a highly penetrant dominant allele. PMID- 18389265 TI - How to store autologous bone graft perioperatively: an in vitro study. AB - INTRODUCTION: Autologous bone graft is the golden standard for bone grafting, but little is known about the influence of various preservation techniques used during surgery immediately following harvest on the osseous structures and graft vitality. Several studies focussed on the optimal treatment of the bone during harvest and implantation, but only few examined the intraoperative storage conditions on the back table. The aim of our study was to examine the influence of various intraoperative preservation techniques on human cancellous bone at different points to optimize the storage during surgery. MATERIALS AND METHODS: Cancellous bone was harvested during hip arthroplasty and stored for 2 and 4 h under dry conditions, inside a swab moistened with saline solution or in saline solution, 5% glucose solution or culture medium. After the storage period, the bone was cultured and examined 7 days after the first cells grew out in one of these groups. Following the identification of the cells as osteoblast-like cells, the cultures were analysed by fluorescence staining, cell count and the XTT-test. RESULTS: Fluorescence staining revealed no avital cells in all groups. Dry storage of the bone led to significantly lower cell metabolism after 2 h compared to saline solution and 5% glucose solution. The same was true after 4 h dry storage compared to the moistened swab, and glucose and culture medium. Cell count was significantly lower after 2 h of dry storage compared to saline solution and culture medium. CONCLUSIONS: Perioperative storage of cancellous bone under dry conditions should be avoided. The bone graft should be stored in saline or 5% glucose solution or a moistened swab. PMID- 18389266 TI - Treatment of congenital scoliosis with single-level hemivertebrae. AB - INTRODUCTION: The natural history of congenital scoliosis with hemivertebrae is unpredictable and the management is also controversial. MATERIALS AND METHODS: Between 1986 and 2004, 22 patients (eight male and fourteen female, mean 19.3 years old) with single-level hemivertebrae related congenital scoliosis underwent non-operative or operative treatment at our institution with an average follow-up period of 8.8 years. RESULTS: Only a 5 degrees curve progression was noted in upper thoracic hemivertebrae after followed up 6 years. By one stage combined anterior hemivertebrae excision, posterior instrumentation, and arthrodesis, up to 61% curve correction can be achieved. Posterior instrumentation, correction and arthrodesis showed a 25% correction. The result of pain relief is promising in skeletal-matured patients. CONCLUSIONS: Surgical instrumentation, correction and arthrodesis showed good results. The optimal treatment of choice may differed from one to the other. PMID- 18389267 TI - Complex level alterations of the 2f (1)-f (2) distortion product due to hypoxia in the guinea pig. AB - It is controversially discussed inasmuch acute hearing disorders might originate from impaired cochlear circulation. Hypoxia-specific alterations of inner ear parameters measurable in patients with acute sensorineural hearing loss would therefore be of great interest. Aim of this study was to characterize hypoxia related alterations of the 2f (1)-f (2) distortion product. Nine guinea pigs were anaesthetized by i.m. administration of Midazolam, Medetomidin and Fentanyl. For introduction of hypoxia, the spontaneously breathing animals were offered a gas mixture of N(2)O and O(2) containing either 21 or 12-13% O(2). Distortion product otoacoustic emissions (DPOAEs) were continuously monitored at f (2) = 16 kHz; f (2)/f (1) = 1, 2; DP-definition = 2f (1)-f (2); L (1) = 65 dB and L (2) = 55 dB, while inhaled oxygen was switched from 21 to 12-13% and back. Oxygen saturation (SaO(2)) was continuously monitored. Data from an hypoxic interval were only used for further data processing if DPOAE levels were stable before and after hypoxia. Six hypoxic intervals in five animals fulfilled the stability criterion. During the hypoxic interval with the highest measured SaO(2) (75%), no alterations of DPOAE levels were observed. During the remaining five hypoxic intervals, when SaO(2) ranged between 57 and 70%, DPOAE levels were on average lower with an increased standard deviation compared to mean pre-hypoxic levels. Mean decrease correlated with the decrease of SaO(2 )(r = 0.90, P = 0.014). Alterations followed a characteristic time course-when hypoxia was started, DPOAE levels exhibited a short increase before they decreased and remarkably destabilized. After re-oxygenation DPOAE levels showed a pronounced level decrease, while SaO(2) already had recovered to pre-hypoxic values. After reaching a minimum, DPOAE levels slowly recovered to pre-hypoxic values. The decrease of DPOAE levels during hypoxia and the post-hypoxic level alterations have similarly been described by other authors before, while the distinct destabilization and transiently increased DPOAE levels have not been explicitly mentioned. A micromechanical mechanism that might explain a transient level increase and the post-hypoxic DPOAE level changes is discussed. PMID- 18389268 TI - The association of smoking, alcoholic consumption, betel quid chewing and oral cavity cancer: a cohort study. AB - We aimed to analyze the relationship between smoking, alcoholic consumption and betel quid chewing with oral cavity cancer. All male patients age > or =18 years who visited our clinic received an oral mucosal inspection. Basic data including personal habits were also obtained. A multivariate logistic regression model was utilized to determine relevant risk factors for developing oral cavity cancer. A total of 8,356 patients were enrolled in this study. Abnormal findings were found in 382 patients (4.6%). Two hundred and ninety-seven patients received biopsy and 191 patients were proven to have oral cavity cancer. The results of multivariate logistic regression showed that those who smoked, consumed alcohol and chewed betel quid on a regular basis were most likely to contract oral cancer (odds ratio: 39.66, 95% confidence interval: 26.04-60.38). Therefore, habitual cigarette smokers, alcohol consumers, and betel quid chewers have a higher risk of contracting oral cavity cancer and should receive oral mucosal screening regularly so potential oral cavity cancer can be detected as early as possible, which may result in better and improved survival of oral cancer patients. PMID- 18389269 TI - Microvascular decompression of cochleovestibular nerve. AB - The role of microvascular decompression (MVD) in the management of trigeminal neuralgia, hemifacial spasms and glossopharyngeal neuralgia is well-established. However, controversy persisted as to the use of MVD in cochleovestibular neurovascular compression syndrome. This report provides a review of all the published studies on MVD of the eighth (8th) nerve in alleviating cochleovestibular symptoms and presents three additional patients who underwent MVD of the eighth nerve for tinnitus or vertigo. Nineteen studies were identified. Five were case reports. The remaining have sample sizes ranging from 4 to 207 patients. Quantitative and qualitative reviews of all studies were performed, focusing on the selection criteria for surgery, efficacy and safety of the procedure. Selection criteria for surgery were variable. No standardised outcome measures were used and all studies rely on patient subjective assessment of surgical outcome. Nonetheless, the results suggest that MVD of the eighth nerve produces good outcome with low morbidity in selected cases. PMID- 18389270 TI - Intercomparison study on (152)Eu gamma ray and (36)Cl AMS measurements for development of the new Hiroshima-Nagasaki Atomic Bomb Dosimetry System 2002 (DS02). AB - In the process of developing a new dosimetry system for atomic bomb survivors in Hiroshima and Nagasaki (DS02), an intercomparison study between (152)Eu and (36)Cl measurements was proposed, to reconcile the discrepancy previously observed in the Hiroshima data between measurements and calculations of thermal neutron activation products. Nine granite samples, exposed to the atomic-bomb radiation in Hiroshima within 1,200 m of the hypocenter, as well as mixed standard solutions containing known amounts of europium and chlorine that were neutron-activated by a (252)Cf source, were used for the intercomparison. Gamma ray spectrometry for (152)Eu was carried out with ultra low-background Ge detectors at the Ogoya Underground Laboratory, Kanazawa University, while three laboratories participated in the (36)Cl measurement using accelerator mass spectrometry (AMS): The Technical University of Munich, Germany, the Lawrence Livermore National Laboratory, USA and the University of Tsukuba, Japan. Measured values for the mixed standard solutions showed good agreement among the participant laboratories. They also agreed well with activation calculations, using the neutron fluences monitored during the (252)Cf irradiation, and the corresponding activation cross-sections taken from the JENDL-3.3 library. The measured-to-calculated ratios obtained were 1.02 for (152)Eu and 0.91-1.02 for (36)Cl, respectively. Similarly, the results of the granite intercomparison indicated good agreement with the DS02 calculation for these samples. An average measured-to-calculated ratio of 0.98 was obtained for all granite intercomparison measurements. The so-called neutron discrepancy that was previously observed and that which included increasing measured-to-calculated ratios for thermal neutron activation products for increasing distances beyond 1,000 m from the hypocenter was not seen in the results of the intercomparison study. The previously claimed discrepancy could be explained by insufficient understanding of the measured data. PMID- 18389271 TI - An X-STR meiosis study in Kurds and Germans: allele frequencies and mutation rates. AB - X-linked short tandem repeats (X-STRs) play an important supplementary role in the field of forensic genetics, especially in deficiency cases. This paper presents population genetic data for the microsatellite markers DXS8378, DXS6800, DXS101, HPRTB, and DXS8377 in a German and a Kurdish population sample. Buccal swabs were obtained from 217 unrelated healthy German individuals (107 women and 110 men) from the area of Munster and 208 unrelated Kurdish individuals (103 women and 105 men), immigrants mainly from Northern Iraq. Additionally, more than 1,200 meiotic transfers (419 paternal and 819 maternal meioses) were investigated in the systems DXS6800, DXS101, and DXS8377. Five mutations were found in the system DXS8377. With the power of discrimination in females [PD(F)] ranging from 0.81 (DXS8378 in Kurds) to 0.99 (DXS8377 in Germans), the investigated X-STRs systems turned out to be highly informative in the two populations. PMID- 18389273 TI - Prevalence of crowded optic discs in adult Chinese. The Beijing Eye Study. AB - BACKGROUND: To determine the prevalence of crowded optic discs and their associations with ocular and general parameters. METHODS: The Beijing Eye Study included 4,439 subjects out of 5,324 subjects invited to participate (response rate 83.4%) with an age of 40+ years. The present investigation consisted of 8,594 eyes (96.8%) of 4,324 subjects (97.4%) for whom readable fundus photographs of at least one eye were available. The main outcome parameter was the presence of crowded optic discs, defined as small discs with an unsharp and slightly prominent disc border without signs of pathology. RESULTS: Crowded optic discs were detected in 265 (3.1 +/- 0.2%) eyes of 168 subjects (84 women). The prevalence rate was 3.8% +/- 0.3% (95% confidence interval: 3.2%, 4.4%) per subject. Crowded optic discs were significantly associated with age (P < 0.05%) and disc size (P < 0.001). They were statistically not associated with gender (P = 0.08), intraocular pressure (P = 0.06), refractive error (P = 0.17), visual field defects (P = 0.53), and best corrected visual acuity (P = 0.57). CONCLUSIONS: Crowded optic discs are present in about 38 out of 1,000 adult Chinese in Northern China, with small optic discs as the main associated factor. These data may be helpful for the assessment of a risk for nonarteritic anterior ischemic optic neuropathy. PMID- 18389272 TI - Refractive properties of the healthy human eye during acute hyperglycemia. AB - PURPOSE: To measure the refractive properties of the healthy human eye during acute hyperglycemia by means of Scheimpflug imaging and Hartmann-Shack aberrometry. METHODS: Acute hyperglycemia was induced in five healthy subjects (two males, three females, mean age +/-SD 24.8 years +/- 4.6) by means of an oral glucose tolerance test (OGTT) after subcutaneous somatostatin injection. Before and every 30 minutes after the OGTT, measurements with Scheimpflug imaging and Hartmann-Shack aberrometry were performed. The main outcome measures were the thickness and shape of the lens, and the ocular refractive error and higher order aberrations. The equivalent refractive index of the lens was calculated from these parameters. Measurements at baseline and during hyperglycemia were analyzed by means of Wilcoxon signed rank sum tests. RESULTS: During hyperglycemia (mean blood glucose level at baseline: 4.0 mmol/l; mean maximal blood glucose level: 18.4 mmol/l) no changes could be found in the refractive properties within the group. In one subject, a hyperopic shift (0.4 D) was observed, together with a more convex shape of the anterior lens surface and a decrease in the equivalent refractive index of the lens. CONCLUSIONS: This study shows that hyperglycemia generally does not cause changes in the refractive properties of the healthy eye. Nevertheless, in one subject a hyperopic shift accompanied by a change in shape and refractive index of the lens was measured. This finding could provide an explanation for the mechanism underlying the refractive changes that are often observed during hyperglycemia. PMID- 18389274 TI - Flap suture--a simple technique for the revision of hypotony maculopathy following trabeculectomy with mitomycin C. AB - BACKGROUND: A serious complication following trabeculectomy with mitomycin C (MMC) is intraocular hypotony with hypotony maculopathy, papilledema, flattening of the anterior chamber, corneal decompensation and a decrease in visual acuity. We describe a new simple surgical technique for the treatment of hypotony maculopathy following trabeculectomy with MMC. METHODS: In a prospective consecutive case study, 16 patients with hypotony maculopathy following trabeculectomy with MMC were included. Through the intact conjunctiva, additional 10.0 nylon sutures were placed through the scleral flaps into the adjacent sclera. Assessment of visual acuity, intraocular pressure (IOP) measurement and fundoscopy were performed on the 1st and 7th day postoperatively and 1, 3 and 6 months postoperatively. Photography of the bleb and the posterior pole and optical coherence tomography imaging of the posterior pole were performed. RESULTS: Before surgery (flap suture), the mean IOP was 2.8 mmHg (+/-1.1; range from 1 to 4 mmHg). It was 24.9 +/- 11.7 mmHg (range; 9-48 mmHg) on the 1st postoperative day, 15.7 +/-8.1 mmHg (range; 5-35 mmHg) on day 7, 11.1 +/-4.1 mmHg (range; 5-20 mmHg) 1 month after surgery, 9.3 +/-3.9 mmHg (range; 2-20 mmHg) after 3 months, and 9.6 +/-4.2 mmHg (range; 2-20 mmHg) after 6 months. The best corrected mean visual acuity was 20/50 before trabeculectomy and 20/160 before flap suture. It improved to 20/63 at 6 months postoperatively. Clinical signs of hypotony maculopathy disappeared in all patients. No bleb leakage was observed in any patient during follow-up. CONCLUSION: Resuturing the scleral flap through the intact conjunctiva is an effective and minimally invasive method to treat hypotony maculopathy following trabeculectomy with MMC. Thus, opening the conjunctiva can be avoided. PMID- 18389275 TI - Quadriceps muscle function characteristics in severely obese and nonobese adolescents. AB - The purpose of this cross-sectional study was to compare quadriceps muscle strength and fatigue between severely obese (body mass index 34 kg/m2) and nonobese adolescents. Maximal isokinetic torque and angle of peak torque as well as isometric torque at short (40 degrees of knee flexion) and long (80 degrees of knee flexion) muscle length were measured using an isokinetic dynamometer. Muscle fatigue was quantified as the percent torque loss during an isokinetic voluntary protocol and an electrical stimulation isometric protocol. Obese adolescents produced greater absolute isokinetic (+16%; P < 0.05) and isometric torque at short (+25%; P < 0.01) but not at long muscle length (P > 0.05) compared to their lean counterparts. The angle of peak torque was significantly lower in obese than in nonobese subjects (-11%; P < 0.05), i.e., obese produced their maximal strength at shorter muscle length. Isokinetic and isometric torque normalized to the fat-free mass were not significantly different between the two groups. No significant difference in voluntary and stimulated torque loss was observed between groups. Muscle strength per unit of fat-free mass and muscle fatigue were similar in the obese and nonobese adolescents tested in this study, therefore suggesting that obesity has little or no effect on quadriceps muscle function characteristics. On the other hand, it remains to be confirmed whether the observed quadriceps muscle length specificity contributes to the reduced functional capacity of obese adolescents during complex motor tasks involving deep knee flexion (squatting, kneeling). PMID- 18389276 TI - Functional involvement of Annexin-2 in cAMP induced AQP2 trafficking. AB - Annexin-2 is required for the apical transport in epithelial cells. In this study, we investigated the involvement of annexin-2 in cAMP-induced aquaporin-2 (AQP2) translocation to the apical membrane in renal cells. We found that the cAMP-elevating agent forskolin increased annexin-2 abundance in the plasma membrane enriched fraction with a parallel decrease in the soluble fraction. Interestingly, forskolin stimulation resulted in annexin-2 enrichment in lipid rafts, suggesting that hormonal stimulation might be responsible for a new configuration of membrane interacting proteins involved in the fusion of AQP2 vesicles to the apical plasma membrane. To investigate the functional involvement of annexin-2 in AQP2 exocytosis, the fusion process between purified AQP2 membrane vesicles and plasma membranes was reconstructed in vitro and monitored by a fluorescence assay. An N-terminal peptide that comprises 14 residues of annexin-2 and that includes the binding site for the calcium binding protein p11 strongly inhibited the fusion process. Preincubation of cells with this annexin-2 peptide also failed to increase the osmotic water permeability in the presence of forskolin in intact cells. Altogether, these data demonstrate that annexin-2 is required for cAMP-induced AQP2 exocytosis in renal cells. PMID- 18389277 TI - Voltage-gated calcium channels in chronic pain: emerging role of alternative splicing. AB - N- and T-type voltage-gated calcium channels are key established players in chronic pain. Current work suggests that alternative splicing of these channels constitutes an important aspect in the investigation of their roles in the pathogenesis of chronic pain. Recent N-type channel studies describe a nociceptor enriched alternatively spliced module responsible for voltage-independent G protein modulation and internalization, which is implicated in the control of distinct nociceptive pathways. On the contrary, although a large body of work has demonstrated that peripheral Cav 3.2-encoded T-type currents are involved in several types of chronic pain, little is known with respect to the expression of numerous newly discovered splice variants in specific pain pathways. The elucidation of the new layers of molecular complexity uncovered in N- and T-type channel splice variants and their respective locations and roles in different pain pathways will allow for the development of better therapeutic strategies for the treatment of chronic pain. PMID- 18389278 TI - The significance of lobular neoplasia on needle core biopsy of the breast. AB - The management of a core biopsy diagnosis of lobular neoplasia is controversial. Detailed radiological-pathological review of 47 patients with cores showing classical lobular neoplasia was performed (patients with pleomorphic lobular carcinoma in situ (LCIS) or associated risk lesions were considered separately). Immediate surgical excision in 25 patients showed invasive carcinoma in 7, ductal carcinoma in situ (DCIS) in 1 and pleomorphic LCIS in 1; radiological pathological review showed that the core biopsy missed a mass in 5, missed calcification in 2 and that calcification appeared adequately sampled in 2. Nineteen patients had follow-up of at least 2 years. Four patients developed malignancy at the site of the core biopsy (invasive carcinoma in three, DCIS in one); one carcinoma was mammographically occult, one patient had dense original mammograms and two had calcifications apparently adequately sampled by the core. In conclusion, most carcinomas identified at the site of core biopsy showing lobular neoplasia were the result of the core missing the radiological lesion, emphasising the importance of multidisciplinary review and investigation of any discordance. Some carcinomas were found after apparently adequate core biopsy, raising the question of whether excision biopsy should be considered after all core biopsy diagnoses of lobular neoplasia. PMID- 18389280 TI - Polymorphisms in the novel serotonin receptor subunit gene HTR3C show different risks for acute chemotherapy-induced vomiting after anthracycline chemotherapy. AB - The aim of this study was to correlate chemotherapy-induced nausea and vomiting (CINV) with commonly occurring single nucleotide polymorphisms (SNP) in the 5 hydroxytryptamine receptor 3 genes (HTR3). Women with breast cancer without previous chemotherapy were eligible for this prospective study. All patients received epirubicin, with or without cyclophosphamide, and preventive medication with ondansetron and dexamethasone. The patients documented every vomiting event on an hourly basis. Real-time polymerase chain reaction (PCR) analysis was performed for the following nonsynonymous SNPs: p.Y129S (HTR3B), p.K163N (HTR3C) and p.A405G (HTR3C). The overall proportion of patients (total n = 110) who reported vomiting in the first 24 h after chemotherapy was 31.8%. The variant genotype of K163N (HTR3C) was associated with vomiting, which occurred in 50.0% (P = 0.009). Polymorphisms in the HTR3C gene could serve as a predictive factor for CINV in patients undergoing moderately emetogenic chemotherapy. PMID- 18389281 TI - Field studies indicating reduced activity of ivermectin on small strongyles in horses on a farm in Central Kentucky. AB - Field studies (n=6) were completed on evaluation of activity of ivermectin (200 microg/kg) paste formulation against small strongyles in horses (foals, yearlings, and older animals) on a farm (Farm MC) in Central Kentucky in late 2006 and during 2007. A girth tape was used to estimate body weights which were then used to calculate the proper dose rate of ivermectin. The foals, yearlings, and some of the older horses were born and raised on the farm. However, most of the older horses which were not raised on the farm had been there for several years. The horse herd was given ivermectin exclusively, usually four times a year, since 1990. An exception was that during the foal's period of life fenbendazole, pyrantel pamoate, and oxibendazole were given occasionally besides ivermectin. Efficacy of drug activity was determined by pretreatment and posttreatment counts of strongyle eggs per gram of feces (EPGs). Culture of strongyle eggs in feces from some of the horses showed that only small strongyle larvae were present. The research included two studies (A and B) in foals (n=24) and four studies (C, D, E, and F) in yearlings (n=13) alone or with older horses (n=10). For each of the studies (B through F), there was a treated and a nontreated group. These groups were switched for each treatment, i.e., the treated group in one study was the nontreated group in the next study and vice versa. Eggs per gram of feces counts were determined at 1- or 2-week posttreatment intervals for 4 weeks for study A and 6 weeks for studies B through F. Also, for studies B, E, and F, counts of EPGs were done either two or three times during the third week posttreatment. The studies showed a similar posttreatment pattern of strongyle EPG counts beginning to return at about 4 weeks and increasing at 5 and 6 weeks posttreatment. Two horses in study E and one in study F had low EPG values toward the end of the third week posttreatment. The results of this ivermectin investigation showed that the strongyle EPG counts started returning about twice as quickly post-ivermectin-treatment of horses than when the drug was first marketed in the early 1980s. PMID- 18389279 TI - What's new in cystic fibrosis? From treating symptoms to correction of the basic defect. AB - Chronic relentless lung infection and pancreatic insufficiency are the cardinal features of cystic fibrosis (CF), a life-shortening autosomal recessive disease. Mutations in the 'cystic fibrosis transmembrane conductance regulator' (CFTR) are currently classified into five groups according to their repercussion on CFTR protein synthesis and its chloride channel function. Stop codon mutations (class I) result in a truncated nonfunctional CFTR, class II mutations consist of aberrantly folded CFTR protein that is degraded by the cell quality control system, while class III mutations lead to defective regulation of the CFTR protein and, consequently, the absence of CFTR function. These three classes usually lead to a classic CF phenotype with pancreatic insufficiency. CFTR mutations that lead to defective chloride conductance are grouped together in class IV. Class V mutations interfere with normal transcription, thereby reducing the amount of otherwise normal CFTR. These latter two classes are mostly associated with a milder expression of the disease. In the absence of CFTR function, unrestrained Na+ absorption and the failure of active Cl- secretion lead to a decreased airway surface liquid (ASL) volume and subsequent failure of normal mucociliary clearance. This review highlights recent therapeutic strategies that either target the underlying defect or the early steps in CF pathophysiology. To date, gene therapy has failed to demonstrate a clinical benefit after repeated administration. Mutation-specific chloride channel correction pharmacotherapy is currently being developed, an example of which is PTC124, a new chemical compound that selectively induces read-through of premature stop codons. However, clinical efficacy for most of the compounds still has to be proven in large clinical trials. The positive effect of nebulised hypertonic saline on mucociliary clearance is based on the restoration of ASL height. Recent advances in the current treatment of lung infection and inflammation are highlighted in this review. Lung transplantation should be considered in terminally ill patients, but the timing of the transplantation is crucial: transplanting too early shortens survival, while transplanting too late results in patients dying on the waiting list. PMID- 18389282 TI - Leishmaniasis treatment--a challenge that remains: a review. AB - Leishmaniasis is a disease caused by flagellate protozoan Leishmania spp. and represents an emergent illness with high morbidity and mortality in the tropics and subtropics. Since the discovery of the first drugs for Leishmaniasis treatment (i.e., pentavalent antimonials), until the current days, the search for substances with antileishmanial activity, without toxic effects, and able to overcome the emergence of drug resistant strains still remains as the current goal. This article reports the development of new chemotherapies through the rational design of new drugs, the use of products derived from microorganisms and plants, and treatments related to immunity as new alternatives for the chemotherapy of leishmaniasis. PMID- 18389283 TI - Outcome after renal transplantation. Part I: intellectual and motor performance. AB - Comprehensive information on neurodevelopmental outcome in children and adolescents with chronic kidney disease is still limited. Intellectual performance (IP) and motor performance (MP) were systematically assessed in 27 patients at a median age of 14.1 years (range 6.5-17) and 6 years (range 0.5 12.7) after renal transplantation (RTPL). IP was analyzed with the Wechsler Intelligence Scale for Children-III (WISC-III) in 25 patients and by the Kaufman Assessment Battery for Children in two patients. MP was evaluated by the Zurich Neuromotor Assessment. Median full- scale intelligent quotient (FSIQ) was 97 (range 49-133). Twenty-one patients had an FSIQ >or= 85 (i.e. >or= mean-1 standard deviation). The five patients with neurological comorbidity had a median FSIQ of 81 (range 49-101). Verbal IQ (VIQ) (median 104; range 50-146) was significantly (p < 0.01) higher than performance IQ (PIQ) (median 88; range 48 117). The PIQ was significantly lower compared with controls (p < 0.007), and patients scored significantly lower compared with controls in five of 11 subtests of the Wechsler Scale. All MP tasks were significantly (p < 0.01) lower than in controls, and also in children without neurological comorbidity. Socioeconomic status was significantly correlated with FSIQ (p = 0.03). IP after RTPL was within the normal range for the majority of children. PIQ was lower compared with VIQ, and MP was significantly affected in all children after RTPL. PMID- 18389284 TI - Childhood chronic kidney disease in a developing country. AB - We have retrospectively reviewed the records of children aged >1 month to 16 years who had been referred to the Department of Pediatrics of Prince of Songkla University's Faculty of Medicine, a tertiary referral center in Thailand, between 1982 and 2005 and subsequently diagnosed with chronic kidney disease (CKD). Our aim was to evaluate the prevalence and etiology of CKD in southern Thailand. There were 101 cases of CKD, with one case each diagnosed in 1988, 1989 and 1993, respectively, and 98 cases diagnosed between 1994 and 2005. These latter cases were divided into two 6-year periods: an early period (1994-1999), with 32 cases, and a later period (2000-2005), with 66 cases. The majority of this pediatric population with CKD were male (62/101, 61.4%). The etiologies of CKD were 35 cases of chronic glomerulonephritis (CGN) (34.7%), 29 of genitourinary tract (GU) anomalies (28.7%), nine of systemic lupus erythematosus (SLE) (8.9%), four malignancies (4.0%), four miscellaneous (4.0%) and 19 of unknown causes (18.8%). The patients were divided into age groups of <2 years (20 CKD patients), 2-6 years (15), >6-10 years (22), >10-13 years (20) and >13 years (24). The etiologies of CKD were significantly different in each age group, with GU anomalies and glomerulonephritis being the major causes of CKD in children aged 6 years (40/65, 61.5%), respectively. In conclusion, the incidence of CKD in our university hospital situation was not rare, with the prevalence doubling during the past 6 years, and the etiologies varying by age group. PMID- 18389285 TI - Spores of the mycorrhizal fungus Glomus mosseae host yeasts that solubilize phosphate and accumulate polyphosphates. AB - The present paper reports the presence of bacteria and yeasts tightly associated with spores of an isolate of Glomus mosseae. Healthy spores were surface disinfected by combining chloramine-T 5%, Tween-40, and cephalexin 2.5 g L(-1) (CTCf). Macerates of these spores were incubated on agar media, microorganisms were isolated, and two yeasts were characterized (EndoGm1, EndoGm11). Both yeasts were able to solubilize low-soluble P sources (Ca and Fe phosphates) and accumulate polyphosphates (polyPs). Sequence analysis of 18S ribosomal deoxyribonucleic acid showed that the yeasts belong to the genera Rhodotorula or Rhodosporidium (EndoGm1) and Cryptococcus (EndoGm11). Results from inoculation experiments showed an effect of the spore-associated yeasts on the root growth of rice, suggesting potential tripartite interactions with mycorrhizal fungi and plants. PMID- 18389286 TI - SAPHO syndrome with rapidly progressing destructive spondylitis: two cases treated surgically. AB - The authors present two cases of synovitis, acne, pustulosis, hyperostosis, osteitis (SAPHO) syndrome with rapidly progressing destructive spondylitis treated surgically. The spinal lesions in SAPHO syndrome generally have a good prognosis and rarely cause the structural destruction or neurological deterioration. Case 1: a 63-year-old female had palmoplantar pustulosis for 2 years. At first, she only felt a pain in the nape with no inducing factor. Two months later, she had incomplete quadriplegia (ASIA scale C). Magnetic resonance imaging showed destruction of C4-C7, kyphotic deformity, and severe compression of the spinal cord. Decompression and reconstruction surgery using anterior and posterior approach improved her paralysis. Case 2: a 69-year-old female complained of persistent back pain. Magnetic resonance imaging revealed spondylitis of T7-T9. Although there were no typical skin lesions, we diagnosed SAPHO syndrome by hyperostosis of the sternocostoclavicular joint and sacral joint. Destruction with kyphotic deformity of the spine progressed gradually for 3 months. Curettage and reconstruction surgery using thoracic endoscope relieved her pain and prevented the destruction of the spine. The histopathology of the specimen obtained surgically showed non-specific inflammation in both cases. Spondylitis in SAPHO syndrome may cause severe destruction and kyphotic deformity followed by paralysis. PMID- 18389287 TI - High glucose promotes collagen synthesis by cultured cells from rat cervical posterior longitudinal ligament via transforming growth factor-beta1. AB - Non-insulin-dependent diabetes is known as a risk factor of ossification of posterior longitudinal ligament, but the mechanism has not been well understood. We hypothesized that hyperglycemia, as a typical characteristic of diabetes, is closely associated with ligament hypertrophy in ossification of posterior longitudinal ligament. In this in vitro study, we investigated the effect of high glucose on collagen synthesis and transforming growth factor-beta1 (TGF-beta1) production using cells isolated from rat cervical posterior longitudinal ligament. The cells were subjected to high D-glucose concentration (25 mM) media for 4 days. Notable increases were observed in gene expression and protein synthesis of collagen types I, III in the cells. The increase was inhibited in the presence of anti-TGF-beta1 antibodies. Production of TGF-beta1 by the cells was also increased significantly by high glucose concentration. Exogenous application of TGF-beta1 was confirmed to increase collagen synthesis of the cells. These data suggested that high glucose could promote collagen synthesis in the posterior longitudinal ligament mainly via endogenous TGF-beta1, resulting in hypertrophy of the ligament. PMID- 18389288 TI - Quality of sleep in patients with chronic low back pain: a case-control study. AB - Animal experiments and studies in humans clearly show that the relation between pain (acute and chronic) and sleep quality is two-way: sleep disorders can increase pain, which in turn may cause sleep disorders. Sleep disorders and chronic low back pain are frequent health problems and it is unsurprising that the two can co-exist. This study was conducted to evaluate if sleep disorders and chronic pain associated are more frequently than one would expect. The objective of the study was to compare sleep quality in a population of patients with chronic low back pain and a control population. Sleep quality was assessed in 101 patients with chronic low back pain (CLBP) and in 97 sex- and age-matched healthy control subjects using the Pittsburgh Sleep Quality Index [PSQI; score from 0 (no disorder) to 21]. The French version of the Dallas Pain Questionnaire (DPQ) was used to assess the impact of low back pain on patients' quality of life. This impact was taken as nil in the healthy controls. The patients with CLBP and the controls were comparable in age, sex, and height, but mean bodyweight was higher in the CLBP group (70.3 +/- 14.5 vs. 61.8 +/- 11.4 kg; P < 0.05). The patients with CLBP were also more frequently on sick leave than the controls (32.3%; n = 31 vs. 0.0% n = 0; P < 0.001). Coffee, tea, and cola intakes were comparable in the two groups. Patients with CLBP had statistically higher scores in all items of the PSQI than the healthy controls. The mean PSQI was 4.7 +/- 3.2 for the healthy controls and 10.9 +/- 7.9 for the patients with CLBP (P < 0.0001). Sleep disorders were greater when the impact of CLBP on daily life (the four aspects of the DPQ) was greater [P < 0.0001]). The sleep of the patients with CLBP was significantly altered compared with that of the healthy controls, in proportion to the impact of low back pain on daily life. Our findings do not indicate whether sleep disorders are a cause or a consequence of CLBP. PMID- 18389289 TI - Relationship between the morphology of the atlanto-occipital joint and the radiographic results in patients with atlanto-axial subluxation due to rheumatoid arthritis. AB - The upper cervical spine is a common focus of destruction from rheumatoid arthritis (RA). Atlanto-axial subluxation (AAS) presents with marked frequency among patients with instability. However, there are occasional patients who show no motion between the occipital bone and atlas on a dynamic cervical radiograph in AAS patients. This study investigated the morphology of the atlanto-occipital joint (AOJ) in AAS patients due to RA using computed tomography, and examined the relationship between its morphology and other radiographic results. Twenty-six consecutive patients with AAS due to RA treated by surgery were reviewed. The subjects included 18 females and 8 males. The average patient age was 59.3 years. The mean duration of RA was 14.3 years. In all the patients, the AOJ was morphologically evaluated using sagittal reconstruction view on computed tomography before surgery. Moreover, the ADI value was investigated at the neutral and maximal flexion position, and atlanto-axial angle (AAA) at the neutral position in preoperative lateral cervical radiographs. The morphology of the AOJ on a CT sagittal reconstruction view was classified into three types as follows: a normal type which showed a maintenance of the joint space, a narrow type which showed a disappearance of the joint space and a fused type which showed the fusion of the AOJ. The pre-operative CT sagittal reconstruction image of the AOJ demonstrated a normal type bilaterally in six cases (Group A). In 15 cases (Group B), CT image demonstrated narrowing on at least one side of the AOJ. In five cases (Group C), CT images demonstrated fusion on at least one side of the AOJ. The average ADI value at the flexion position was 10.7 mm in Group A, 11.7 mm in Group B, and 12.6 mm in Group C. There was no significant difference among those groups. The average ADI value at the neutral position before surgery was 2.8 mm in Group A, 5.9 mm in Group B, and 10.4 mm in Group C. There was no significant difference between Group A and B (P > 0.105), and Groups B and C (P > 0.032), however, there was a significant difference between Groups A and C (P < 0.004). The average AAA value was 25.3 degrees in Group A, 19.3 degrees in Group B and 3.4 degrees in Group C. There was no significant difference between Groups A and B (P > 0.230), however, there was a significant difference between Groups A and C (P < 0.002), and Groups B and C (P < 0.007). This study showed that fusion or ankylosis of the AOJ induced an enlargement of the ADI and anterior inclination of the atlas in the neutral position, despite the fact that normal findings of AOJ showed a slight displacement of the atlas to axis in RA patients showing AAS involvement. This morphology may progress to SAS and VS due to AOJ after atlanto-axial arthrodesis. PMID- 18389290 TI - Intramedullary hemangioblastomas: timing of surgery, microsurgical technique and follow-up in 23 patients. AB - Hemangioblastomas are highly vascularised tumors of the central nervous system and account for 1.5-2.5% of all spinal cord tumors. Because of the rarity of these tumors, surgical experience is often limited and, therefore, treatment and indications for timing of surgery are discussed controversial. The authors reviewed their data of 23 consecutive patients with respect to timing of surgery, microsurgical technique, and follow-up. Clinical records of 23 consecutive patients with intramedullary hemangioblastomas who underwent first surgery in our department between 1990 and 2005 were reviewed. In three cases the tumors were localised at the craniocervical junction; four patients had a single tumor in the cervical spine, six patients multiple tumors in the cervical and thoracic spine, eight patients in the thoracic spine only, one patient in the conus region, and one patient had multiple tumors located in the thoracic and lumbar spine. In eight patients, a von-Hippel-Lindau disease (VHL) was associated. The neurological follow-up was evaluated according to the classification of McCormick. Operation was recommended to every symptomatic patient as early as possible. In asymptomatic patients with a sporadic tumor surgery was discussed for diagnostic purposes at any time. In VHL patients, surgery was recommended if tumor growth was observed on MRI in the next practicable time. All tumors were diagnosed by magnetic resonance imaging and in all cases but one a DSA was performed. All patients were treated microsurgically through a posterior approach. The tumors in the spinal cord were removed microsurgically through a partial hemilaminectomy (n = 1), a hemilaminectomy (n = 15), or laminectomy (n = 4) and at the craniocervical junction (n = 3) through a suboccipital craniotomy. During follow-up after 6 months, 18 patients remained neurologically stable (17 in McCormick grade I and 1 in McCormick grade II) and 5 patients recovered to a better status (3 from grade III to II, 2 from grade II to I). There was one complication with a CSF fistula and one recurrence/incomplete removal. Following the above-mentioned principles of microsurgical removal of intramedullary hemangioblastomas, operation is possible with a low procedure-related morbidity and can be recommended especially in VHL patients with progressive symptoms or tumor growth during follow-up. Patients without VHL most frequently require hemangioblastoma resection for diagnostic purposes and/or because symptoms prompted an imaging work-up that lead to the discovery of the tumor. PMID- 18389291 TI - Comparison of open versus percutaneous pedicle screw insertion in a sheep model. AB - Minimally invasive surgery has become more and more important for the treatment of traumatic spine fractures. Besides, some clinical studies, objective data regarding the possible lower damage to the surrounding tissue of the spine is still missing. Here we report a sheep model where we compared a percutaneous versus an open approach for dorsal instrumentation with pedicle screws to the spine. Twelve skeletally mature sheep underwent bilateral pedicle screw fixation at the L4-L6 level. Forty-eight pedicle screws were bilaterally inserted into the pedicles and connected with rods using either an open dorsal standard or a percutaneous approach. Operation time, blood flow, compartment pressure, radiation time, loss of blood, laboratory findings and EMG were evaluated to objectify possible advantages for the percutaneous operation technique. Loss of blood and the distribution of CK-MM as a marker for muscle damage were significantly lower in the percutaneous group. However, radiation time was significantly longer in the percutaneous group. Other parameters like compartment pressure, blood flow and also measurement of the EMG at different time points did not reveal significant differences. Based on the results we found in the present study, percutaneous screw insertion can bring moderate advantages but it should be noted that essential functional deficits to the muscle could not be detected. PMID- 18389292 TI - Association of bone morphogenetic protein-2 gene polymorphisms with susceptibility to ossification of the posterior longitudinal ligament of the spine and its severity in Chinese patients. AB - A case-control study was conducted to examine the association between two single nucleotide polymorphisms (SNPs) in exon 2 of the bone morphogenetic protein-2 gene (BMP-2) and ossification of the posterior longitudinal ligament (OPLL), and to investigate whether SNPs of the Ser37Ala (T/G) and the Ser87Ser (A/G) in the BMP-2 gene are associated with genetic susceptibility to OPLL and its severity in Chinese subjects. The Ser87Ser (A/G) SNP has been implicated in bone mineral density (BMD) and increases the risk of OA in women. The Ser37Ala (T/G) SNP is associated with BMD and the rate of bone loss in osteoporosis and osteoporosis fractures. A total of 57 OPLL patients and 135 non-OPLL controls were studied. Radiographs of the cervical spine were analyzed to determine the presence and the severity of OPLL. The association of two SNPs with the occurrence and the extent of OPLL were statistically evaluated. There was a significant association between the Ser37Ala (T/G) polymorphism and the occurrence of OPLL in the cervical spine. However, no significant association was found between the Ser37Ala (T/G) polymorphism and the more number of ossified cervical vertebrae in OPLL patients. There was a significant association between the Ser87Ser (A/G) polymorphism and the more number of ossified cervical vertebrae in OPLL patients. However, there was no statistical difference between the Ser87Ser (A/G) SNP and the occurrence of OPLL in the cervical spine. In addition, the Ser87Ser (A/G) polymorphism in male patients and in female patients showed no statistical difference between cases and controls. The present results demonstrate that BMP-2 Gene is not only a factor associated with the occurrence of OPLL, but also a factor related to more extensive OPLL. The "G" allele in the Ser37Ala (T/G) polymorphism is associated with the occurrence of OPLL, but not more extensive OPLL in the cervical spine. The "G" allele in the Ser87Ser (A/G) polymorphism promotes the extent of OPLL, whereas the "A" allele in the Ser87Ser (A/G) polymorphism restricts ectopic ossification in the cervical spine at least in Chinese subjects. PMID- 18389293 TI - Comments about "European guidelines for the diagnosis and treatment of pelvic girdle pain". PMID- 18389294 TI - Donor site morbidity following iliac crest bone harvesting for cervical fusion: a comparison between minimally invasive and open techniques. AB - We have studied the occurrence of donor site morbidity, cosmesis and overall satisfaction with graft procedure in 76 patients who had undergone iliac crest bone harvesting for anterior cervical discectomy and fusion (ACDF). Totally 24 patients underwent an open procedure and 52 a minimally invasive trephine harvesting method. Although our study demonstrated substantial donor site pain and its effect on ambulation in both groups, this was of limited duration. Two patients, one in each group, suffered long-term pain that was eventually resolved. Totally 8.3% of patients in the open group suffered minor complications and 11.5% in the trephine group. There were two cases of meralgia parasthetica. There were no major complications in either group. There was no statistically significant difference in morbidity between the open and trephine groups. There was a trend towards significance (P = 0.076) for pain at the donor site, with less pain reported by patients who underwent the trephine procedure for harvesting. PMID- 18389295 TI - Spinal synovial cysts: pathogenesis, diagnosis and surgical treatment in a series of seven cases and literature review. AB - This study is designed based on the retrospective analysis of patients treated in the Neurosurgical Department of two major hospitals and review of the literature. The aim of this study is to evaluate the efficacy of surgery and address controversial issues in the treatment of symptomatic lumbar intraspinal synovial cysts. Spinal juxtafacet cysts (synovial and ganglion cysts) are a rare cause of low back and radicular leg pain. Although the relevant reports in the international literature are increasing, the controversy about conservative versus surgical treatment and the need for concomitant fusion still exists. Data from seven patients (age range 58-69 years, mean age 61 years) with low back and radicular leg pain due to a lumbar facet joint cyst were retrospectively analyzed. Demographic data, cyst level, presence of concomitant local pathology, treatment and results of treatment were recorded. A follow-up of at least 6 months (range 6-48 months) was conducted and results were noted. All patients had back pain, while five also experienced unilateral radicular leg pain and one had bilateral leg pain. One patient had neurogenic claudication. MRI identified the cyst and highlighted underlying pathology in all cases. All patients underwent surgical cyst excision. No fusion was performed. Post-operatively, all patients showed a total resolution of symptoms with sustained benefit at final evaluation. Review of the literature revealed a trend towards surgery, as this is correlated to a more favorable outcome compared with conservative treatment. Fusion should be performed on a case-by-case basis only. Surgery is a safe and effective treatment choice in this increasingly appearing ailment. A prospective, randomized trial should clarify issues under debate. PMID- 18389297 TI - Development of a stable continuous flow immobilized enzyme reactor for the hydrolysis of inulin. AB - A 23.5-fold purified exoinulinase with a specific activity of 413 IU/mg and covalently immobilized on Duolite A568 has been used for the development of a continuous flow immobilized enzyme reactor for the hydrolysis of inulin. In a packed bed reactor containing 72 IU of exoinulinase from Kluyveromyces marxianus YS-1, inulin solution (5%, pH 5.5) with a flow rate of 4 mL/h was completely hydrolyzed at 55 degrees C. The reactor was run continuously for 75 days and its experimental half-life was 72 days under the optimized operational conditions. The volumetric productivity and fructose yield of the reactor were 44.5 g reducing sugars/L/h and 53.3 g/L, respectively. The hydrolyzed product was a mixture of fructose (95.8%) and glucose (4.2%) having an average fructose/glucose ratio of 24. An attempt has also been made to substitute pure inulin with raw Asparagus racemosus inulin to determine the operational stability of the developed reactor. The system remained operational only for 11 days, where 85.9% hydrolysis of raw inulin was achieved. PMID- 18389296 TI - The metal-binding features of the recombinant mussel Mytilus edulis MT-10-IV metallothionein. AB - In contrast with the paradigmatic mammalian metallothioneins (MTs), mollusc MT systems consist at least of a high-cadmium induced form, possibly involved in detoxification, and another isoform either constitutive or regulated by essential metals and probably associated with housekeeping metabolism. With the aim of providing a deeper characterization of the coordination features of a molluscan MT peptide of the latter kind, we have analyzed here the metal-binding abilities of the recombinant MeMT-10-IV isoform of Mytilus edulis (MeMT). Also, comparison with other MTs of this type has been undertaken. A synthetic complementary DNA was constructed, cloned and expressed into two Escherichia coli systems. Upon zinc coordination, MeMT folds in vivo into highly chiral and stable Zn(7) complexes, with an exceptional reluctance to fully substitute cadmium(II) and/or copper(I) for zinc(II). In vivo cadmium binding leads to homometallic Cd(7) complexes that structurally differ from any of the in vitro prepared Cd(7) complexes. Homometallic Cu-MeMT can only be obtained in vitro from Zn(7)-MeMT after a great molar excess of copper(I) has been added. In vivo, two different heterometallic Zn,Cu-MeMT complexes are recovered, which nicely correspond to two distinct stages of the in vitro zinc/copper replacement. These MeMT metal-binding features are consistent with a physiological role related to basal/housekeeping metal, mainly zinc, metabolism, and confirm the correspondence between the MeMT gene response pattern and the functional properties of the encoded protein. PMID- 18389298 TI - Seawater requirement for the production of lipoxazolidinones by marine actinomycete strain NPS8920. AB - A novel marine actinomycete strain NPS8920 produces a new class of 4 oxazolidinone antibiotics lipoxazolidinone A, B and C. Lipoxazolidinone A possesses good potency (1-2 microg/mL) against drug-resistant pathogens methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecium (VRE). Strain NPS8920 exhibits different morphologies in both agar and submerged cultures. The ability of strain NPS8920 to sporulate on saline-based agar media but not on deionized water-based agar medium supported that strain NPS8920 is a marine actinomycete. While strain NPS8920 does not require seawater for growth, the production of lipoxazolidinones by strain NPS8920 can only be detected in the seawater-based media. The optimal production of lipoxazolidinones was observed in the natural seawater-based medium. Strain NPS8920 produced 10-20% of lipoxazolidinones in the synthetic sea salt Instant Ocean-based medium and no production in the sodium chloride-based and deionized water-based media. PMID- 18389299 TI - Increased postprandial triglyceride-rich lipoprotein levels in elderly survivors of myocardial infarction. AB - Postprandial triglyceride-rich lipoproteins (TRL) levels are a predictor for coronary atherosclerosis. The aim of the study was to compare fasting high density lipoprotein (HDL) cholesterol, plasma lipoprotein lipase (LPL) activity, and postprandial TRL between elderly survivors of myocardial infarction (MI) and healthy controls. A case-control study was performed in 44 elderly patients 65-85 years of age with a previous history of MI and 43 age- and sex-matched healthy controls. Each participant underwent physical examination and was given a standard oral fat load with subsequent blood sampling over the next 8 h. Total and chylomicron triglycerides were assessed by area under the curve (AUC), incremental are under the curve (AUCi) and triglyceride response (TGR). Elderly MI patients had significantly lower postheparin LPL activity (87.4 +/- 36.9 mU/ml) (mean +/- 1 SD) than healthy controls (106.0 +/- 29.0 mU/ml) (P = 0.014). Decreased postheparin LPL activity was accompanied by significant increased and delayed clearance of postprandial TRL. Fasting HDL cholesterol was significantly lower in elderly MI patients than controls (1.45 +/- 0.32 and 1.66 +/- 0.47 mmol/l, respectively, P = 0.048). Multiple regression analysis revealed postheparin LPL activity as an independent predictor for postprandial TRL and fasting HDL cholesterol. Logistic regressions analysis revealed HDL cholesterol, triglycerides measured 2 h after the oral fat load, and postheparin LPL activity as independent predictors for MI. Our findings indicate that decreased fasting HDL cholesterol is associated with increased postprandial triglyceridemia which could be a target for life-style and therapeutic interventions in patients at risk for cardiovascular disease. PMID- 18389300 TI - [Analgesic intolerance (AI). Key position of ENT physicians for early detection of this condition]. AB - The clinical manifestation of analgesic intolerance (AI) is frequently associated with symptoms examined by ear-nose-throat (ENT) specialists. The prevalence of AI is reported in the literature to be 0.6-2.5%. Even though there are no concluding results concerning its pathogenesis, an altered arachidonic acid metabolism is most likely the underlying pathomechanism. The symptoms include chronic rhinosinusitis with nasal polyps, asthma, gastrointestinal ulcers, angioedema, and urticaria. Clinical reactions after ingestion of nonsteroidal anti inflammatory-drugs (NSAIDs) are often obvious in the progress of disease. In order to initiate early therapy and therefore prevent the progress of disease, the diagnosis of AI should occur before the complete picture of AI is obvious. Adaptive desensitization is currently the single causal therapy. Frequency of endonasal revision surgery is reduced after desensitization; severe asthma and reactions after ingestion of NSAIDs are avoided. ENT specialists are particularly in a key position for early detection of AI. PMID- 18389302 TI - Rethinking proteasome evolution: two novel bacterial proteasomes. AB - The proteasome is a multisubunit structure that degrades proteins. Protein degradation is an essential component of regulation because proteins can become misfolded, damaged, or unnecessary. Proteasomes and their homologues vary greatly in complexity: from HslV (heat shock locus v), which is encoded by 1 gene in bacteria, to the eukaryotic 20S proteasome, which is encoded by more than 14 genes. Despite this variation in complexity, all the proteasomes are composed of homologous subunits. We searched 238 complete bacterial genomes for structures related to the proteasome and found evidence of two novel groups of bacterial proteasomes. The first, which we name Anbu, is sparsely distributed among cyanobacteria and proteobacteria. We hypothesize that Anbu must be very ancient because of its distribution within the cyanobacteria, and that it has been lost in many more recent species. We also present evidence for a fourth type of bacterial proteasome found in a few beta-proteobacteria, which we call beta proteobacteria proteasome homologue (BPH). Sequence and structural analyses show that Anbu and BPH are both distinct from known bacterial proteasomes but have homologous structures. Anbu is encoded by one gene, so we postulate a duplication of Anbu created the 20S proteasome. Anbu's function appears to be related to transglutaminase activity, not the general stress response associated with HslV. We have found different combinations of Anbu, BPH, and HslV within these bacterial genomes, which raises questions about specialized protein degradation systems. PMID- 18389303 TI - Surgical procedure for reversal of nipple piercing. AB - Body piercing of the nipples for adornment with jewelry has increased worldwide. Many patients later regret their decision and request reversal along with a desire to maintain their ability to breast feed. The surgeon must remove the epithelial tunnel with minimum damage to adjacent ducts. The simple technique described uses the patient's jewelry post as a guide and a common disposable biopsy punch. Slid over the post, the punch excises the epithelial tunnel core with minimal harm to surrounding structures. PMID- 18389304 TI - Flector tissugel used to treat capsular contracture after breast augmentation surgery. AB - Capsular contracture constitutes the main postoperative complication after breast augmentation by implant placement. To date, no systemic treatment known allows for improvement that does not simultaneously put the patient at risk for secondary complications of a more general nature. Flector Tissugel is the sole locally active antiinflammatory patch. Its durable local antiinflammatory effect is associated only with a risk for rare and highly limited side effects. After approximately 3 weeks of treatment, a high frequency of change from capsular contracture Baker 2 or 3 to Baker 1 occurs, provided the application was started no later than 3 months after the onset of capsular contracture. PMID- 18389305 TI - Diffusion of policy discourse into rural spheres through co-management of state forestlands: two cases from West Java, Indonesia. AB - In the context of state forestland management in tropical regions, the implementation of a co-management approach has been widely advocated in order to include the voices of local people and accommodate their interests in management decision-making. Most co-management literatures, however, underestimate the significance of statutory authority held by state to control forestlands and resources. By clarifying the implications of state ownership of forestland, this article aims to critically examine co-management processes with reference to Foucault's notion of power and subject. Case studies were conducted at two co management pilot sites in Gunung Halimun-Salak National Park, West Java, Indonesia. Findings demonstrate that co-management processes actually materialize shared decision-making arrangements between state forest bureaucracy and rural people through the application of equity approaches, such as deliberation, negotiation, and experimentation. At the same time, these processes can also function to diffuse state policy discourse in rural spheres, which makes rural subjects who accept and practice the policy discourse. The research also reveals that the diffusion process is complex and does not necessarily make a durable subject unless they are pertinently organized. The results of this research indicate that co-management of state forestlands is a double-edged process for local people who risk becoming a proxy of state bureaucracy in the implementation of state policy. Proponents of co-management should, therefore, critically examine whether new institutional arrangements, which are developed through co management, truly reflect values and needs of local people and assist them to develop a pertinent subject to deal with it. PMID- 18389306 TI - Establishing green roof infrastructure through environmental policy instruments. AB - Traditional construction practices provide little opportunity for environmental remediation to occur in urban areas. As concerns for environmental improvement in urban areas become more prevalent, innovative practices which create ecosystem services and ecologically functional land cover in cities will be in higher demand. Green roofs are a prime example of one of these practices. The past decade has seen the North American green roof industry rapidly expand through international green roof conferences, demonstration sites, case studies, and scientific research. This study evaluates existing international and North American green roof policies at the federal, municipal, and community levels. Green roof policies fall into a number of general categories, including direct and indirect regulation, direct and indirect financial incentives, and funding of demonstration or research projects. Advantages and disadvantages of each category are discussed. Salient features and a list of prompting standards common to successfully implemented green roof strategies are then distilled from these existing policies. By combining these features with data collected from an experimental green roof site in Athens, Georgia, the planning and regulatory framework for widespread green roof infrastructure can be developed. The authors propose policy instruments be multi-faceted and spatially focused, and also propose the following recommendations: (1) Identification of green roof overlay zones with specifications for green roofs built in these zones. This spatial analysis is important for prioritizing areas of the jurisdiction where green roofs will most efficiently function; (2) Offer financial incentives in the form of density credits and stormwater utility fee credits to help overcome the barriers to entry of the new technology; (3) Construct demonstration projects and institutionalize a commitment greening roofs on publicly-owned buildings as an effective way of establishing an educated roofing industry and experienced installers for future green roof construction. PMID- 18389307 TI - Bio-reduction of selenite to elemental red selenium by Tetrathiobacter kashmirensis. AB - A bacterium that detoxifies selenite by reduction to insoluble elemental red selenium was isolated from soil. The strain showed an unusually high resistance to the toxic effects of selenite by growing in media containing 64 mM selenite. 16S rRNA gene sequence alignment identified the isolate as Tetrathiobacter kashmirensis. Fatty acid analysis and morphology confirmed the identification. The isolate reduced selenite to elemental selenium under aerobic conditions only. Native gel electrophoresis of cell-free extracts revealed a band, corresponding to a molecular weight of approximately 120 kDa, that reduced selenite. In culture, the strain did not reduce selenate; however, a soluble and inducible enzyme with a molecular weight of approximately 90 kDa that reduced both selenate and nitrate was present in cell-free extracts. This organism might be useful in bioreactors designed to remove selenite from contaminated water. PMID- 18389308 TI - Immobilization of alginate-encapsulated Bacillus thuringiensis var. israelensis containing different multivalent counterions for mosquito control. AB - Immobilized techniques have been used widely for the controlled release formulation of mosquitoes. Among the microbial formulations, polymeric matrices play an important role in the controlled release of microbial pesticide at rates sufficiently effective to kill mosquitoes in the field. The advantage of these matrices is that they enhance the stability of both spores and toxin against pH, temperature variations, and UV irradiation. The disadvantage of using calcium alginate beads is that they are unstable upon contact with phosphate of potassium or sodium ions rich in the mosquito habitats. To overcome these problems, attempts were made to encapsulate Bacillus thuringiensis var. israelensis within alginate by using different multivalent counterions, namely, calcium chloride, zinc sulfate, copper sulfate, cobalt chloride, and ferric chloride, and the beads formed were tested for its mosquito larvicidal activity. Among all the beads tested, zinc alginate beads resulted in maximum larvicidal activity of 98% (+/ 1.40 SE) against Culex quinquefasciatus IIIrd instar larvae and maximum spore count of 3.36 x 10(5) (+/-5291.50 SE) CFU/ml. Zinc alginate beads maintained their structure for up to 48 h when shaken vigorously on a rotary shaker at 180 rpm in the presence of 10 mM potassium phosphate buffer (pH 6.8 +/- 0.1). In conclusion, our results suggest that the use of zinc sulfate as counterions to encapsulate B. thuringiensis var. israelensis within alginate may be a potent mosquito control program in the habitats where more phosphate ions are present. PMID- 18389309 TI - Resistance of Deinococcus radiodurans to mutagenesis is facilitated by pentose phosphate pathway in the mutS1 mutant background. AB - MutS1 is a key protein involved in mismatch repair system for ensuring fidelity of replication and recombination in Deinococcus radiodurans. The zwf gene encodes glucose-6-phosphate dehydrogenase (G6PD) in the pentose phosphate (PP) pathway, which provides adequate metabolites as precursors of DNA repair. In this study, mutS1 and zwf were disrupted by homologous recombination. The zwf mutant (Deltazwf) and the zwf/mutS1 double mutant (Deltazwf/mutS1) were sensitive to ultraviolet (UV) light, H(2)O(2), and DNA cross-linking agent mitomycin C (MMC), whereas the mutS1 mutant (DeltamutS1) showed resistance to UV light, H(2)O(2) and MMC as the wild-type strain. Inactivation of mutS1 resulted in a 3.3-fold increase in frequency of spontaneous rifampicin-resistant mutagenesis and a 4.9 fold increment in integration efficiency of a donor point-mutation marker during bacterial transformation. Although inactivation of zwf had no obvious effect compared with the wild-type strain, dual disruption of zwf and mutS1 resulted in a 4.7-fold increase in mutation frequency and a 7.4-fold increase in integration efficiency. These results suggest that inactivation of the PP pathway decreases the resistance of D. radiodurans cells to DNA damaging agents and increases mutation frequency and integration efficiency in the mutS1 mutant background. PMID- 18389310 TI - Effects of ambroxol on alginate of mature Pseudomonas aeruginosa biofilms. AB - Biofilm-forming bacteria Pseudomonas aeruginosa is a common pathogen in mechanically ventilated newborns, which can cause life-threatening infections. Alginate of mucoid Pseudomonas aeruginosa biofilms is considered an important virulence factor which contributes to the resistance to antibiotics. Traditionally, ambroxol is widely used in newborns with lung problems as a mucolytic agent and antioxidant agent as well. And there are few studies that demonstrated the anti-biofilm activity of ambroxol. In this study, we found that ambroxol can affect the structure of mucoid Pseudomonas aeruginosa biofilms. Further, we found that ambroxol reduces the production of alginate, the expression of the important genes and the activity of key enzyme guanosine diphospho-D-mannose dehydrogenase (GDP-mannose dehydrogenase; GMD) which were involved in alginate biosynthesis. PMID- 18389311 TI - Functional analysis of the Campylobacter jejuni cj0183 and cj0588 genes. AB - The cj0183 and cj0588 genes identified in the Campylobacter jejuni NCTC 11168 genome encode proteins with amino acid sequences predicted to be homologous to other bacterial hemolysins. The Cj0183 protein exhibits homology to Brachyspira hyodysenteriae TlyC protein, whereas the cj0588 gene product is homologous to TlyA proteins Brachyspira hyodysenteriae, Helicobacter pylori, and Mycobacterium tuberculosis, which play a crucial role in bacterial virulence. The aim of our work was to examine the hemolytic activity and determine the role of cj0183- and cj0588-encoded proteins on the adherence of chosen C. jejuni strains to the Caco 2 cell line by constructing deletion mutants in the mentioned genes. We found out there is no difference in hemolytic activity between both mutants in gene cj0183 and cj0588 and the wild strains. However, Cj0588 protein but not Cj0183 is involved in adherence to the Caco-2 cells. PMID- 18389312 TI - Minimally invasive surgery for colorectal cancer: quality of life, body image, cosmesis, and functional results. AB - BACKGROUND: The aims of this cross-sectional study were to assess the long-term quality of life, the body image, and the cosmetic and functional results in patients who had laparoscopic-assisted for colorectal cancer. METHODS: Forty-two patients were enrolled in this study: 21 consecutive patients who had undergone laparoscopic-assisted colonic resection and 21 patients who had open colonic resection, selected according to stage, gender, age, cancer site, and type of resection. The patients answered four questionnaires about their quality of life, body image, functional, and cosmetic results. Nonparametric tests were used for statistical analysis. RESULTS: Postoperative hospital stay was shorter in patients who had laparoscopic-assisted resection. The cosmetic score was significantly better in the laparoscopic-assisted group than in the open group (p < 0.01). In spite of similar overall body image score, patients who had a laparoscopic-assisted resection reported a significantly better satisfaction with their own body (p = 0.05). Quality-of-life and functional results were similar in both groups. CONCLUSIONS: The cosmetic results and the consequent satisfaction with the body were significantly better after laparoscopic assisted resection compared to equivalent open procedure. These effects seemed to be temporary but they could help patients to accept the burden of surgery. PMID- 18389313 TI - Mechanical versus suture fixation of the port in adjustable gastric banding procedures: a prospective randomized blinded study. AB - BACKGROUND: Laparoscopic adjustable gastric band port placement can require 9-25% of total implantation time. Prolonged port fixation combined with extended wound manipulation results in an elevated incidence of port-related complications (infection, port tension, pain, rotation). This study compared port fixation using a novel applier to traditional fixation using nonresorbable sutures. METHODS: From Dec 2005 to March 2006, 50 patients were enrolled into a prospective randomized study. Intra- and postoperative outcomes of placing the Swedish Adjustable Gastric Band((R)) (SAGB) Realize with Velocity port and fixation device (group A) were compared to those of the SAGB with its titanium suture-fixation port (group B). Patient characteristics, port implantation duration, incision length, postoperative port-site pain [via visual analogue scale (VAS)], pain medication, and complications were recorded. Minimum follow-up was 12 months (three visits). Postoperative evaluation was performed by an investigator not involved in the surgery. RESULTS: Patient groups were comparable in age, sex, and body mass index. In group A versus group B, mean port implantation time was lower (53.5 +/- 13.1 versus 451.3 +/- 84.9 s, p < 0.0001), port-site skin incision was longer (mean 4.1 +/- 0.2 versus 3.1 +/- 0.2 cm, p < 0.0001), and postoperative opioid consumption was less (mean 1.3 versus 3.0 units, p < 0.01). Group A reported less mean pain on the VAS (3.2 versus 3.8, p < 0.01); 0/25 group A versus 3/25 group B patients required hypnotics. There were no discernible differences between groups in hospitalization (mean 2.9 days), nursing requirements, or pain at 3, 6, and 12 months. In group A, no patient required reoperation for a port complication; two port rotations occurred in group B (NS). CONCLUSION: Velocity mechanical port applier fixation is superior in efficiency to port suture fixation, safely increasing the speed to port implant from nearly 8 min to under 1 min and decreasing total operative time by 19%. Port applier fixation trends to reduction of major complications and statistically significantly lessens the minor complication of port-site pain. PMID- 18389314 TI - Use of a colonoscope for distal duodenal stent placement in patients with malignant obstruction. AB - BACKGROUND: Stent placement in the distal duodenum or proximal jejunum with a therapeutic gastroscope can be difficult, because of the reach of the endoscope, loop formation in the stomach, and flexibility of the gastroscope. The use of a colonoscope may overcome these problems. OBJECTIVE: To report our experience with distal duodenal stent placement in 16 patients using a colonoscope. METHODS: Multicenter, retrospective series of patients with a malignant obstruction at the level of the distal duodenum and proximal jejunum and treated by stent placement using a colonoscope. Main outcome measurements are technical success, ability to eat, complications, and survival. RESULTS: Stent placement was technically feasible in 93% (15/16) of patients. Food intake improved from a median gastric outlet obstruction scoring system (GOOSS) score of 1 (no oral intake) to 3 (soft solids) (p = 0.001). Severe complications were not observed. One patient had persistent obstructive symptoms presumably due to motility problems. Recurrent obstructive symptoms were caused by tissue/tumor ingrowth through the stent mesh [n = 6 (38%)] and stent occlusion by debris [n = 1 (6%)]. Reinterventions included additional stent placement [n = 5 (31%)], gastrojejunostomy [n = 2 (12%)], and endoscopic stent cleansing [n = 1 (6%)]. Median survival was 153 days. CONCLUSION: Duodenal stent placement can effectively and safely be performed using a colonoscope in patients with an obstruction at the level of the distal duodenum or proximal jejunum. A colonoscope has the advantage that it is long enough and offers good endoscopic stiffness, which avoids looping in the stomach. PMID- 18389315 TI - Delayed laparoscopic subtotal cholecystectomy in acute cholecystitis with severe fibrotic adhesions. AB - BACKGROUND: Conversion rate to open surgery is higher for patients with acute cholecystitis than in those without acute cholecystitis. We attempted to develop a laparoscopic subtotal cholecystectomy to decrease this conversion rate. METHODS: From 2000 to 2005, laparoscopic cholecystectomy for acute cholecystitis was performed in 60 patients (22 women, 38 men). Patients were divided into two groups: group A (2000 to 2002, n = 22) and group B (2003 to 2005, n = 38). When significant difficulty was encountered dissecting the gallbladder from its bed, we incised the gallbladder wall leaving the posterior wall and cauterizing the remnant mucosa (subtotal cholecystectomy, SC-1). When dissection of the gall bladder neck and triangle of Calot was difficult, the neck of the gallbladder was sutured despite clipping (SC-2). RESULTS: Mean duration from onset of symptoms to operation was 55.3 +/- 52.0 days. SC-1 was performed in 8 patients in group A and 18 patients in group B. SC-2 was performed in three patients in Group B. Conversion rate was 18.1% (4/22) in group A and 0% (0/38) in group B, compared to 0.4% (1/221) for patients without acute cholecystitis. No complications were associated with ablated gallbladder mucosa. CONCLUSION: Laparoscopic subtotal cholecystectomy offers safe and effective treatment for acute cholecystitis. The conversion rate in group B is decreased by avoiding hazardous dissection of the cystic duct. PMID- 18389316 TI - Clinical models are inaccurate in predicting bile duct stones in situ for patients with gallbladder. AB - BACKGROUND: The probability that a patient has common bile duct stones (CBDS) is a key factor in determining diagnostic and treatment strategies. This prospective cohort study evaluated the accuracy of clinical models in predicting CBDS for patients who will undergo cholecystectomy for lithiasis. METHODS: From October 2005 until September 2006, 335 consecutive patients with symptoms of gallstone disease underwent cholecystectomy. Statistical analysis was performed on prospective patient data obtained at the time of first presentation to the hospital. Demonstrable CBDS at the time of endoscopic retrograde cholangiopancreatography (ERCP) or intraoperative cholangiography (IOC) was considered the gold standard for the presence of CBDS. RESULTS: Common bile duct stones were demonstrated in 53 patients. For 35 patients, ERCP was performed, with successful stone clearance in 24 of 30 patients who had proven CBDS. In 29 patients, IOC showed CBDS, which were managed successfully via laparoscopic common bile duct exploration, with stone extraction at the time of cholecystectomy. Prospective validation of the existing model for CBDS resulted in a predictive accuracy rate of 73%. The new model showed a predictive accuracy rate of 79%. CONCLUSION: Clinical models are inaccurate in predicting CBDS in patients with cholelithiasis. Management strategies should be based on the local availability of therapeutic expertise. PMID- 18389317 TI - Correlation of CO2 pneumoperitoneal pressures between rodents and humans. AB - INTRODUCTION: Many studies have reported on the effects of pneumoperitoneum in a rat model, using a wide range of intra-abdominal pressures. The correlation between pneumoperitoneal pressures in rodents to pressures in humans has not been established. This study evaluates changes in various physiological parameters in different pneumoperitoneum pressures in the rat model with a comparison to known data in humans. MATERIALS AND METHODS: Three groups of eight Wistar rats each were anesthetized, tracheostomized, and mechanically ventilated with fixed tidal volume and respiratory rate. After a stabilization phase, CO(2) pneumoperitoneum was established to 12, 8, and 5 mmHg in the different groups. Changes in blood pressure, heart rate, peak ventilatory pressure, and end-tidal CO(2) (ETCO(2)) were recorded throughout the experiment. RESULTS: There were no significant changes in blood pressure and heart rate in all groups. No increase in ETCO(2) was demonstrated following induction of pneumoperitoneum in the 12 and 8 mmHg groups. A statistical significant increase in ETCO(2) occurred only in the 5 mmHg group (39.4 to 41.3 mmHg, p = 0.023). Ventilatory pressures increased after induction of pneumoperitoneum in all groups. The increase reached a maximal level in the 8 and 12 mmHg groups (from 3 to 12 mmHg) and was lower in the 5 mmHg group (from 3 to 7 mmHg) CONCLUSIONS: The neglected increase in ETCO(2) in pressures > or =8 mmHg, in the rat correlates to high pressures in humans (above 14-20 mmHg) when CO(2) diffusion through the peritoneum declines due to pressure occlusion of peritoneal capillaries. The maximal ventilatory pressures generated in the rat in intra-abdominal pressures > or =8 mmHg correlate to pressures, which are higher than the standard working pressures in humans. Thus, pneumoperitoneal pressures >8 mmHg in the rat do not simulate routine working pressures employed in humans. A pressure of 5 mmHg is optimal in a rat model to simulate laparoscopy in humans. PMID- 18389318 TI - Cytokine responses following laparoscopic or open pyeloplasty in children. AB - BACKGROUND: The present study evaluated the cytokine response in children following laparoscopic pyeloplasty (LP) or open pyeloplasty (OP). A series of cytokines were measured postoperatively, including interkin1-beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-alpha), and C-reactive protein (CRP). METHODS: A total of 31 patients, with an average age of 9.1 +/- 3.0 years (range 2.5-14 years) were studied. Fourteen patients underwent LP and 17 underwent OP. Blood serum concentrations of IL-1beta, IL-6, IL-8, IL-10, TNF-alpha, and CRP were measured via enzyme-linked immunosorbent assay (ELISA) before surgery as well as 4, 24, and 48 h following the operation. In addition, the procedure duration, hospital stay, incidence of wound infection, and the recurrence rate of stenosis in both groups were compared. RESULTS: Serum IL-6 and CRP concentrations were significantly elevated in both groups at 4, 24, and 48 h relative to preoperative levels. However, the rise in IL-6 and CRP in OP group was significantly more robust than in LP group. No significant changes were observed in serum levels of IL-1beta, IL-8, IL-10, or TNF-alpha in either group. The procedure duration was significantly longer for LP (193.6 +/- 74.7 min, range 120-360 min) versus OP (120.1 +/- 27.5 min, range 90-165 min, p < 0.05), but the hospital stay following LP was shorter (LP group: 5.3 +/- 1.1 days versus OP group: 9.3 +/- 2.1 days, p < 0.05). No severe complications were noted in either group, however, one child experienced wound infection following OP procedure. An incident of recurrent stenosis following the operation occurred in both groups. There was no postoperative morbidity or severe implications at 12 month follow-up in either group. CONCLUSIONS: Both OP and LP are safe and effective procedures for the treatment of ureteropelvic junction obstruction in the pediatric population. However, the shorter hospital stay and decreased cytokine response following LP indicates potential benefits over traditional invasive procedures. PMID- 18389319 TI - Incorporation of proficiency criteria for basic laparoscopic skills training: how does it work? AB - BACKGROUND: It is desirable that surgical trainees are proficient in basic laparoscopic motor skills (eye-hand coordination). The present study evaluated the use of predefined proficiency criteria on a basic virtual reality (VR) simulator in preparation for a laparoscopic course on animal models. METHODS: Twenty-eight surgical trainees who enrolled for a basic laparoscopic course were trained on a basic (VR) simulator until their performance met predefined criteria. Two different criteria were defined, based on the performance of experienced laparoscopic surgeons on the simulator. In the first group (n = 10), the criteria were set at the 75th percentile of the laparoscopic surgeons' performance on the simulator and in the second group, at the 50th percentile (n = 18). Training time and number of attempts needed until the performance criteria were met were measured. RESULTS: In the first group, training time needed to pass the test ranged from 29 to 77 min (median: 63 min) with a range of 43-90 attempts (median 61 attempts). In the second group, training time ranged from 38 to 180 min (median 80 min) with a range of 55-233 attempts (median 95 attempts). Experience with assisting or performing laparoscopic procedures varied widely and was not correlated with the training time and number of attempts needed to pass the criteria. CONCLUSIONS: The performance criteria for training laparoscopic motor skills on a (VR) simulator resulted in wide variation between surgical trainees in time and number of attempts needed to pass the criteria. This demands training courses with a flexible time span tailored to the individual level of the trainee. PMID- 18389320 TI - Diagnostic laparoscopy guidelines : This guideline was prepared by the SAGES Guidelines Committee and reviewed and approved by the Board of Governors of the Society of American Gastrointestinal and Endoscopic Surgeons (SAGES), November 2007. PMID- 18389322 TI - An interval >7 weeks between neoadjuvant therapy and surgery improves pathologic complete response and disease-free survival in patients with locally advanced rectal cancer. AB - BACKGROUND: We assessed whether the time interval between neoadjuvant therapy and surgery affects the operative and postoperative morbidity and mortality, the pathologic complete response (pCR) rate, and disease recurrence in locally advanced rectal cancer. METHODS: One-hundred and thirty-two patients with locally advanced low- and mid-rectal cancer underwent neoadjuvant chemoradiation followed by radical resection (October 2000 to December 2006). Data on the neoadjuvant regime, neoadjuvant-surgery interval, final pathology, type of operation, operative time, intraoperative blood transfusions, postoperative complications, length of hospital stay, disease recurrence, and mortality were reviewed. The patients were divided into two groups according to the neoadjuvant-surgery interval: 7 weeks (group B, n = 84). RESULTS: The groups were demographically comparable except for the group A patients being younger at operation. The median interval between chemoradiation and surgery was 56 days (range 13-173 days). Thirty-seven patients (28%) had a pCR and near pCR. Fifty three patients (40%) had complications. There was no in-hospital mortality. Surgery type, operative time, number of intraoperative blood transfusions, postoperative complications, and length of hospitalization were not influenced by the interval length. The pCR and near pCR rates were higher with longer interval: 17% in group A, 35% in group B (P = 0.03). Patients operated at an interval >7 weeks had significantly better disease-free survival (P = 0.05). CONCLUSIONS: A neoadjuvant-surgery interval >7 weeks was associated with higher rates of pCR and near pCR, decreased recurrence and improved disease-free survival. PMID- 18389321 TI - Radical resection after IORT-containing multimodality treatment is the most important determinant for outcome in patients treated for locally recurrent rectal cancer. AB - BACKGROUND: The optimal treatment for locally recurrent rectal cancer (LRRC) is still a matter of debate. This study assessed the outcome of LRRC patients treated with multimodality treatment, consisting of neoadjuvant radio (chemo-) therapy, extended resection, and intraoperative radiotherapy. METHODS: One hundred and forty-seven consecutive patients with LRRC who underwent treatment between 1994 and 2006 were studied. The prognostic values of patient-, tumor- and treatment-related characteristics were tested with uni- and multivariate analysis. RESULTS: Median overall survival was 28 months (range 0-146 months). Five-year overall, disease-free, and metastasis-free survival and local control (OS, DFS, MFS, and LC respectively) were 31.5%, 34.1%, 49.5% and 54.1% respectively. Radical resection (R0) was obtained in 84 patients (57.2%), microscopically irradical resection (R1) in 34 patients (23.1%), and macroscopically irradical resection (R2) in 29 patients (19.7%). For patients with a radical resection median OS was 59 months and the 5-year OS, DFS, MFS, and LC were 48.4%, 52.3%, 65.5% and 68.9%, respectively. Radical resection was significantly correlated with improved OS, DFS, and LC (P < 0.001). Patients who received re-irradiation or full-course radiotherapy survived significantly longer (P = 0.043) and longer without local recurrence (P = 0.038) or metastasis (P < 0.001) compared to patients who were not re-irradiated. CONCLUSIONS: Radical resection is the most significant predictor of improved survival in patients with LRRC. Neoadjuvant radio (chemo-) therapy is the best option in order to realize a radical resection. Re-irradiation is feasible in patients who already received irradiation as part of the primary rectal cancer treatment. PMID- 18389323 TI - Criteria in sentinel lymph nodes of melanoma patients that predict involvement of nonsentinel lymph nodes. AB - BACKGROUND: Previous studies described various criteria in sentinel lymph nodes (SLN) of melanoma patients that predict the involvement of further, nonsentinel lymph nodes (NSLN). Such criteria may facilitate the selection of patients who might benefit from a completion lymph node dissection (CLND). However, it is currently unclear which parameters are most important. METHODS: A total of 180 melanoma patients with positive SLNB and subsequent CLND were investigated. Histopathologic parameters in the SLN were systematically evaluated and compared with regard to NSLN positivity. Twenty-eight of these patients (16.0%) had positive NSLN. RESULTS: By univariate analysis several criteria with regard to tumor burden and location of melanoma cells in the SLN correlated with NSLN involvement, such as positivity by hematoxylin-eosin (H&E) staining (P < .001), largest diameter of clusters (P < .001), capsular involvement (P = .001), extranodal extension (P < .001), and tumor penetrative depth (P < .001). Multivariate analysis revealed three independent parameters: (1) positivity of the SLN by H&E staining (versus by immunohistochemistry alone), (2) relative tumor burden > 10% of total lymph node tissue, and (3) perinodal intralymphatic tumor. In 23 of 28 patients with positive NSLN the SLN was positive by H&E staining, in 15 of 28 patients the relative tumor burden was > 10%, and 13 of 28 showed perinodal intralymphatic tumor. In 5 of 28 patients with NSLN involvement, these three parameters were negative. CONCLUSIONS: Histopathologic examination of the SLN can identify patients at risk for NSLN positivity. PMID- 18389324 TI - Enhancing the informal curriculum of a medical school: a case study in organizational culture change. AB - BACKGROUND: Calls for organizational culture change are audible in many health care discourses today, including those focused on medical education, patient safety, service quality, and translational research. In spite of many efforts, traditional "top-down" approaches to changing culture and relational patterns in organizations often disappoint. OBJECTIVE: In an effort to better align our informal curriculum with our formal competency-based curriculum, Indiana University School of Medicine (IUSM) initiated a school-wide culture change project using an alternative, participatory approach that built on the interests, strengths, and values of IUSM individuals and microsystems. APPROACH: Employing a strategy of "emergent design," we began by gathering and presenting stories of IUSM's culture at its best to foster mindfulness of positive relational patterns already present in the IUSM environment. We then tracked and supported new initiatives stimulated by dissemination of the stories. RESULTS: The vision of a new IUSM culture combined with the initial narrative intervention have prompted significant unanticipated shifts in ordinary activities and behavior, including a redesigned admissions process, new relational practices at faculty meetings, student-initiated publications, and modifications of major administrative projects such as department chair performance reviews and mission-based management. Students' satisfaction with their educational experience rose sharply from historical patterns, and reflective narratives describe significant changes in the work and learning environment. CONCLUSIONS: This case study of emergent change in a medical school's informal curriculum illustrates the efficacy of novel approaches to organizational development. Large-scale change can be promoted with an emergent, non-prescriptive strategy, an appreciative perspective, and focused and sustained attention to everyday relational patterns. PMID- 18389325 TI - Duodenoduodenal intussusception. AB - Duodenoduodenal intussusception is a rare event which is usually caused by the presence of a tumor. We present a case of duodenoduodenal intussusception secondary to a large tubulovillous adenoma causing gastric outlet and biliary obstruction in a 50-year-old female. The imaging features on ultrasonography, CT, and MRI are described. PMID- 18389329 TI - Chemoprevention by white currant is mediated by the reduction of nuclear beta catenin and NF-kappaB levels in Min mice adenomas. AB - BACKGROUND: Berries are a good natural source of phenolic compounds and many berries or their compounds have been shown to be chemopreventive. White currant is an interesting berry, as it contains low levels of dominant berry phenolics such as ellagic acid, anthocyanins and other flavonoids. AIMS OF THE STUDY: To study if white currant is chemopreventive in an experimental model for intestinal tumorigenesis and further study the effects on beta-catenin and NF-kappaB signaling pathways. METHODS: Multiple intestinal neoplasia (Min) mice were fed an AIN-93G based control diet or a diet containing 10% freeze dried white currant (Ribes x pallidum) for 10 weeks. Cell signaling parameters were analysed from intestinal adenomas and surrounding mucosa by Western blotting and immunohistochemistry. RESULTS: The white currant diet reduced the number of adenomas from 81 (min-max 47-114) to 51 (36-84) in the total small intestine of Min mice (P<0.02). Most of the adenomas develop in the distal part of the small intestine, and in this area white currant reduced the number from 49 to 29.5 (P<0.01) and also the size of the adenomas from 0.88 mm to 0.70 mm (P<0.02). In the colon white currant increased the number of adenomas (0.3+/-0.6 vs. 0.8+/ 0.6, mean +/- SD, P<0.05), but did not affect the size. White currant reduced nuclear beta-catenin and NF-kappaB protein levels in the adenomas (P<0.05 and P<0.02, respectively). They were correlated with the size of adenomas (P<0.01). CONCLUSIONS: This study shows that white currant is effective in preventing cancer initiation and progression in the Min mouse. Whether the positive effects are due to its special phenolic composition needs to be studied in more detail. PMID- 18389330 TI - Effects of drug-eluting stents on systemic inflammatory response in patients with unstable angina pectoris undergoing percutaneous coronary intervention. AB - Inflammatory markers are elevated in acute coronary syndromes, and are also known to play a crucial role in the pathogenesis of neointimal proliferation and stent restenosis. Drug-eluting stents (DESs) have been shown to decrease stent restenosis in different studies. In this study, we aimed to investigate the effect of treatment with DESs on systemic inflammatory response in patients with unstable angina pectoris who underwent percutaneous coronary intervention (PCI). We compared plasma high-sensitivity C-reactive protein (hsCRP), human tumor necrosis factor alpha (Hu TNF-alpha), and interleukin 6 (IL-6) levels after DES (dexamethasone-eluting stent [DEXES], and sirolimuseluting stent [SES]) implantation with levels after bare metal stent (BMS) implantation. We performed PCI with a single stent in 90 patients (62 men; 59 +/- 9 years of age; n = 30 in the BMS group, n = 30 in the DEXES group, n = 30 in the SES group) who had acute coronary syndrome. Plasma hsCRP, Hu TNF-alpha, and IL-6 levels were determined before intervention and at 24 h, 48 h, and 1 week after PCI. The results were as follows. Plasma hsCRP levels at 48 h (11.19 +/- 4.54, 6.43 +/- 1.63 vs 6.23 +/- 2.69 mg/l, P = 0.001) after stent implantation were significantly higher in the BMS group than in the DES group; this effect persisted for 7 days (P = 0.001). Plasma Hu TNF-alpha levels at each time point were higher in the SES group than in the BMS and DEXES groups (P < 0.05). The time course of Hu TNF-alpha values was similar in all groups. Although IL-6 levels at baseline and at 24 and 48 h showed no statistically significant difference between the study groups, postprocedural values at 7 days were slightly statistically significant in the SES group (P = 0.045). Drug-eluting stents showed significantly lower plasma hsCRP levels after PCI compared with BMSs. This may reflect the potent effects of DESs on acute inflammatory reactions induced by PCI. PMID- 18389331 TI - Comparison of effect between nitrates and calcium channel antagonist on vascular function in patients with normal or mildly diseased coronary arteries. AB - The comparative long-term antianginal efficacy of long-acting nitrates versus calcium channel antagonists remains unclear. The goal of the present study was to compare the coronary endothelial cell function and coronary artery vasoconstriction between patients with normal or mildly diseased coronary arteries treated with long-acting nitrates or calcium channel antagonists. Forty two patients suspected to have angina pectoris and with normal or mildly diseased coronary arteries underwent Doppler flow study of the left anterior descending coronary artery. All patients were suspected to have angina pectoris and were receiving either long-acting nitrates (n = 18; Nitrates group) or calcium channel antagonists (n = 24; Ca-antagonists group) for at least 1 year. Vascular reactivity was assessed by intracoronary administration of papaverine, acetylcholine (Ach), and nitroglycerin using a Doppler guidewire. Segments that showed the greatest constrictive response to Ach were used for assessment of vasoconstriction. The percent increase in coronary blood flow (CBF) and coronary artery diameter (CAD) induced by Ach was significantly smaller in the Nitrates group than in the Ca-antagonists group (33% +/- 74% vs 83% +/- 77%, P < 0.05; -3% +/- 16% vs 11% +/- 12%, P < 0.01, respectively). The percent diameter reduction in the region of greatest constrictive response to Ach was significantly greater in the Nitrates group than in the Ca-antagonists group (44% +/- 39% vs 15% +/- 32%, P < 0.02). Long-term treatment with long-acting nitrates may produce less favorable effects on coronary endothelial function and the constrictive response to Ach when compared with long-acting calcium channel antagonists in patients with normal or mildly diseased coronary arteries. PMID- 18389332 TI - Synergistic effect of amlodipine and atorvastatin on blood pressure, left ventricular remodeling, and C-reactive protein in hypertensive patients with primary hypercholesterolemia. AB - Our aim in this study was to investigate the changes of serum high-sensitive C reactive protein (hs-CRP) and uric acid (UA), and evaluate the synergistic effect of amlodipine and atorvastatin on blood pressure and left ventricular remodeling in hypertensive patients with primary hypercholesterolemia. One hundred and twenty-six hypertensive patients with hypercholesterolemia were randomized into amlodipine group (10 mg/day, group A, n = 65) and amlodipine (10 mg/day) plus atorvastatin group (20 mg/day, group B, n = 61), treated for 4 months continuously. Serum concentrations of total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, hs-CRP, and UA were determined, and blood pressure of both groups was examined before and after treatment. Left ventricular posterior wall thickness and interventricular spectum thickness were measured by echocardiography, and left ventricular mass index (LVMI) was calculated. After 4-months of treatment with atorvastatin, serum concentrations of total cholesterol, low-density lipoprotein cholesterol, triglycerides, hs-CRP, and UA were significantly decreased in group B (P < 0.05, P < 0.01), while serum concentrations of high-density lipoprotein cholesterol was elevated (P < 0.05). Meanwhile, systolic blood pressure and diastolic blood pressure were reduced in both groups (P < 0.05), and blood pressure in group B was markedly lower than that in group A after treatment (P < 0.05). Compared with that before treatment, LVMI in both groups decreased (P < 0.05), to a significantly lower degree in group B than in group A (P < 0.05). Atorvastatin can decrease serum concentrations of hs-CRP and UA. The amlodipine-atorvastatin combination markedly reduces blood pressure and reverses left ventricular hypertrophy more than amlodipine monotherapy. The positive effect suggests that in hypertensive and hypercholesterolemic patients, the combination of amlodipine and atorvastatin could be the treatment of choice. PMID- 18389333 TI - Abdominal obesity and inflammation predicts hypertension among prehypertensive men and women: the ATTICA Study. AB - The aim of this work was to assess the 5-year incidence of hypertension and its predictors among prehypertensive adults. Under the context of the ATTICA Study, data from 1188 individuals, free of cardiovascular disease, but with defined high blood pressure levels (prehypertension) at baseline examination (during 2001 2002) were retrieved. In 2006, the 5-year follow-up of the study was performed, and 798 of the prehypertensive participants were allocated. In this work, incidence and determinants of developing hypertension were evaluated. The 5-year ageadjusted incidence of hypertension was 18.7% in men and 24.6% in women (P = 0.05); while almost one half of prehypertensive individuals at the age of 55-65 years developed hypertension, and approximately 6 out of 10 people over 65 years of age developed the disease. Multiple logistic regression analysis revealed that increased age (odds ratio [OR] per 1 year = 1.09, 95% confidence interval [CI] 1.07-1.12), male sex (OR = 0.40, 95% CI 0.21-0.68), high education status (OR per 1 year of school = 0.94, 95% CI 0.88-0.98), waist circumference (OR per 1 cm = 1.04, 95% CI 1.02-1.06) and C-reactive protein (OR per 1 mg/l = 1.12, 95% CI 1.05 1.20), were positively associated with the development of hypertension. Moreover, greater adherence to Mediterranean diet seems to protect only prehypertensive, with abdominal obesity patients prone to develop hypertension (OR = 0.94, 95% CI 0.90-0.98). Annual incidence of hypertension was roughly 4% in men and women. Older people, with low education, abdominal obesity, lower adherence to the Mediterranean diet, and increased inflammation, constitute a model of prehypertensive individuals that are prone to develop hypertension. PMID- 18389334 TI - The K121Q polymorphism of ENPP1 and peripheral arterial disease. AB - The K121Q variant of the ectonucleotide pyrophosphatase/ phosphodiesterase 1 (ENPP1) gene is associated with obesity, insulin resistance, and early myocardial infarction. Therefore, we hypothesized that the K121Q polymorphism might also be associated with an increased risk for peripheral arterial disease. Four hundred patients with peripheral arterial disease and 400 controls matched for sex and age (+/- 2 years) were genotyped cross-sectionally for the K121Q single nucleotide polymorphism of the ENPP1 gene. The frequency for the 121Q allele was 0.25 both in patients with peripheral arterial disease and in controls (P = 0.75). Subgroup analysis revealed association of the ENPP1 121QQ genotype with higher glycohemoglobin A1C levels (P = 0.001) and earlier onset of peripheral arterial disease (P = 0.003) in the cohort of nonsmokers. Whereas the K121Q genotype of the ENPP1 gene is not associated with presence of peripheral arterial disease in the whole Linz Peripheral Arterial Disease population, it is associated with type 2 diabetes mellitus and earlier onset of peripheral arterial disease in the subgroup of nonsmoking patients. PMID- 18389335 TI - The mild form of mucopolysaccharidosis type I (Scheie syndrome) is associated with increased ascending aortic stiffness. AB - Mucopolysaccharidosis type I (MPS IS) is a rare autosomal recessive disease caused by a deficiency of the lysosomal enzyme alpha-L-iduronidase, which is involved in the degradation of sulfated glycosaminoglycans (GAGs). The deficiency results in the intra-and pericellular accumulation of the GAGs heparan sulfate and dermatan sulfate. Eight adult patients with typical features of MPS IS aged 31.5 +/- 6.8 years (five men) were included and compared to age-and gender matched controls. With transthoracic echocardiography, cyclic ascending aortic diameter changes were measured and ascending aortic elastic properties were calculated to characterize aortic elasticity. In MPS IS patients, aortic stiffness index was significantly increased (23.1 +/- 10.4 vs 3.9 +/- 1.5, P < 0.001), while aortic distensibility was significantly decreased (1.6 +/- 0.8 vs 4.6 [corrected] +/- 1.9 cm(2)/dynes [corrected] 10(-6), P < 0.001) compared to age-and sex-matched controls. The results of the present study demonstrate that in addition to the known cardiac complications, MPS IS patients have an impairment of ascending aortic elasticity. Further follow-up studies are needed to examine arterial elasticity using other methods in this patient population, and to detect possible effects of enzyme replacement therapy. PMID- 18389336 TI - Functional implications of the right ventricular myocardial performance index in patients after surgical repair of tetralogy of Fallot. AB - The myocardial performance index (MPI) has been proposed to be a simple echocardiographic index of right ventricular (RV) function in patients after surgical repair of tetralogy of Fallot (TOF). However, its functional status remains to be clarified. The functional implications of RV MPI were determined by exploring its relationships with parameters of RV function as derived from cardiovascular magnetic resonance (CMR), and exercise capacity of postoperative TOF patients Thirty patients (11 males), aged 15.6 +/- 3.1 years, who have undergone surgical repair of TOF at 4.0 +/- 1.8 years, were studied. The RV and left ventricular (LV) MPIs determined using pulsed-wave Doppler echocardiography were related to CMR-derived RV and LV ejection fractions, and pulmonary regurgitant fraction and treadmill exercise testing parameters. Log RV MPI correlated positively with log LV MPI (r = 0.38, P = 0.037) and negatively with CMR-derived RV ejection fraction (r = -0.4, P = 0.028) and pulmonary regurgitant fraction (r = -0.4, P = 0.031). No significant correlations were found between LV MPI and any of the CMR parameters. Using receiver operated characteristics analysis, a cutoff value of 0.30 for RV MPI was found to have a sensitivity of 100% and specificity of 74% in predicting a RV ejection fraction <35%. Right ventricular, but not LV, MPI correlated inversely with exercise duration (r = 0.45, P = 0.013) and peak oxygen consumption (VO2 max) (r = -0.56, P = 0.001). Multivariate analysis identified RV MPI (beta = -0.6, P < 0.001), male sex (beta = 0.44, P = 0.01), and duration from surgery (beta = -0.30, P = 0.019) as significant determinants of VO2 max. Increased MPI is a reflection of reduced RV ejection fraction and exercise capacity in patients after TOF repair. PMID- 18389337 TI - Visualization of the azygos arch valves on multidetector-row computed tomography. AB - To evaluate the frequency and appearance of the azygos arch valves on chest examinations using multidetector-row computed tomography (MDCT), we retrospectively reviewed findings from 194 contrast-enhanced MDCT examinations of the chest. Rate of injection of 300 mgI/ml contrast materials was low (2.0 ml/s) and high (3.0 ml/s). Scanning delay was 80 s on examination on low-rate injection of contrast material and 20 s on high-rate injection of contrast material. The presence of residual contrast material in the azygos arch valves and reflux of contrast material into the azygos arch were recorded. The Cochran-Armitage trend test was used to compare the frequency of residual contrast material in the azygos arch valves and reflux of contrast material into the azygos arch in both groups. Of 92 examinations of high-rate injection of contrast material, 63 (68.5%) demonstrated residual contrast material in the azygos arch valves and 71 (77.2%) demonstrated reflux of contrast material into the azygos arch. A significantly higher frequency of reflux of contrast material into the azygos arch and residual contrast material in the azygos arch valves was seen in the high-rate injection group than in the low-rate injection group (P < 0.05). Residual contrast material in the azygos arch valves was demonstrated more frequently when contrast material was administered in the right side of the arm than in the left side of the arm (P < 0.05). Reflux of contrast material into the azygos arch was common in the high-injection-rate group and residual contrast material in the azygos arch valves was far more frequently seen in the high injection-rate group than in the low-injection-rate group on MDCT. PMID- 18389339 TI - Electron paramagnetic resonance investigation on modulatory effect of benidipine on membrane fluidity of erythrocytes in essential hypertension. AB - It has been shown that benidipine, a long-lasting calcium (Ca) channel blocker, may exert its protective effect against vascular disorders by increasing nitric oxide (NO) production. The purpose of the present study was to investigate whether orally administered benidipine might influence the membrane function in patients with essential hypertension. We measured the membrane fluidity of erythrocytes by using an electron paramagnetic resonance (EPR) and spin-labeling method. In the preliminary study using erythrocytes obtained from healthy volunteers, benidipine decreased the order parameter (S) for 5-nitroxide stearate (5-NS) and the peak height ratio (ho/h-1) for 16-NS in the EPR spectra in vitro. The finding indicated that benidipine increased the membrane fluidity and improved the microviscosity of erythrocytes. In addition, it was demonstrated that the effect of benidipine on membrane fluidity of erythrocytes was significantly potentiated by the NO-substrate, L-arginine. In the separate series of the study, we observed that orally administered benidipine for 4 weeks significantly increased the membrane fluidity of erythrocytes with a concomitant increase in plasma NO metabolite levels in hypertensive subjects. The results of the present study demonstrated that benidipine might increase the membrane fluidity and improve the microviscosity of erythrocytes both in vitro and in vivo, to some extent, by the NO-dependent mechanism. Furthermore, it is strongly suggested that orally administered benidipine might have a beneficial effect on the rheologic behavior of erythrocytes and the improvement of the microcirculation in hypertensive subjects. PMID- 18389338 TI - Beneficial effects of low doses of red wine consumption on perturbed shear stress induced atherogenesis. AB - Moderate wine intake is associated with a reduced risk of morbidity and mortality from cardiovascular disease. Atherosclerosis is enhanced in arterial segments exposed to disturbed flow. Perturbed shear stress increases also the endothelial expression of oxidation-sensitive responsive genes (such as ELK-1 and p-JUN). This study evaluates the effects of chronic consumption of red wine on perturbed shear stress-induced atherogenesis. Results indicated that chronic treatment with red wine significantly attenuated the activation of redox-sensitive genes (ELK-1 and p-JUN) and increased endothelial nitric oxide synthase (eNOS) expression (which was decreased by perturbed shear stress) in cultured human coronary endothelial cells (EC) and in atherosclerosis-prone areas of hypercholesterolemic mice. Oral administration of red wine to hypercholesterolemic mice reduced significantly the progression of atherosclerosis. Moreover, short-term supplementation with red wine to C57BL/6J mice significantly increased upregulation of aortic eNOS and SIRT1 expression induced by physical training. These findings establish that administration of low doses of red wine can attenuate the proatherogenic effects induced by perturbed shear stress in vitro and in vivo. This evidence may have implications for the prevention of atherosclerotic lesion progression and its clinical manifestations. PMID- 18389340 TI - Aberrant right coronary artery occlusion during the percutaneous pulmonary trunk stenting in a patient with tetralogy of Fallot. AB - Aberrant coronary arteries are frequently observed in patients presenting with Fallot's tetralogy (TOF). Before the complete surgical repair of the TOF, the percutaneously performed pulmonary trunk (PT) angioplasty is often performed in order to temporarily increase the pulmonary circulation, thus increasing the pulmonary vessel size, finally improving surgical outcome. This case reports a 12 year-old boy with a TOF insufficiently improved by surgical correction, in whom a PT angioplasty with stent implantation was complicated by an extrinsic compression of an aberrant right coronary artery (RCA) causing a myocardial ischemia with severe hypotension. The RCA, originating from the left anterior descending coronary artery, passed through the aortic root and the PT and was thus compressed by the PT-stent. Finally the RCA was successfully treated with standard coronary balloon angioplasty and stenting, improving myocardial perfusion and the hemodynamics of the patient, who finally died several days thereafter due to septic shock and massive pulmonary embolism. PMID- 18389341 TI - A novel COL3A1 gene mutation in patient with aortic dissected aneurysm and cervical artery dissections. AB - A considerable proportion of aortic aneurysms are shown to have genetic backgrounds. The COL3A1 gene, which encodes type III procollagen and causes Ehlers-Danlos syndrome (EDS) type IV, is one of the candidate genes associated with aortic aneurysms. The COL3A1 gene is also associated with cervical artery dissections (CAD) mostly as a part of manifestations of EDS type IV. We describe a 34-year-old Korean woman with both abdominal dissected aortic aneurysm and CAD accompanying atrial septal defect and multiple cysts in ovary and thyroid glands. She lacked cardinal manifestations of EDS type IV other than the vascular abnormalities, but molecular analyses of the COL3A1 gene confirmed a de novo heterozygous missense mutation that has not been reported before (c. 781G > A; Gly261Ser). This case suggested that the COL3A1 gene could be one of etiologically linked genes in isolated vasculopathies such as aortic dissected aneurysm or CAD, although being rare. PMID- 18389342 TI - The interaction of zinc oxide-based dental cements with aqueous solutions of potassium fluoride. AB - The ability of zinc oxide-based dental cements (zinc phosphate and zinc polycarboxylate) to take up fluoride from aqueous solution has been studied. Only zinc phosphate cement was found to take up any measurable fluoride after 5 h exposure to the solutions. The zinc oxide filler of the zinc phosphate also failed to take up fluoride from solution. The key interaction for this uptake was thus shown to involve the phosphate groups of the set cement. However, whether this took the form of phosphate/fluoride exchange, or the formation of oxyfluoro phosphate groups was not clear. Fluoride uptake followed radicaltime kinetics for about 2 h in some cases, but was generally better modelled by the Elovich equation, dq(t)/dt = alpha exp(-betaq(t)). Values for alpha varied from 3.80 to 2.48 x 10(4), and for beta from 7.19 x 10(-3) to 0.1946, though only beta showed any sort of trend, becoming smaller with increasing fluoride concentration. Fluoride was released from the zinc phosphate cements in processes that were diffusion based up to M(t)/M(infinity) of about 0.4. No further release occurred when specimens were placed in fresh volumes of deionised water. Only a fraction of the fluoride taken up was re-released, demonstrating that most of the fluoride taken up becomes irreversibly bound within the cement. PMID- 18389343 TI - Biodistribution of titanium dioxide from biologic compartments. AB - The layer of titanium dioxide (TiO(2)) of the implant is chronically exposed to the internal electrolyte milieu in the peri-implant biological compartment. Corrosion results from electrochemical attack and ensuing gradual degradation of the metallic materials and is thus of biological interest when these biomaterials are employed in clinical implantology. Herein we evaluated and compared the chronic effect and the biodistribution of TiO(2) administered subcutaneously or intraperitoneally. We propose that the compartmentalization of titanium in the area of subcutaneous injection would reproduce the biological compartment of the implant and its microenvironment from which metal ions could be released and migrate systemically. Potential TiO(2) deposits were identified and characterized in skin, liver and lung by histological and EDX analyses. After both treatments, the skin, liver, and lungs exhibited histological evidence of TiO(2) deposits. In order to characterize in situ macrophage-like cells, tissue sections were immunohistochemically stained for CD68. Tissue specimens from all organs assayed showed positive staining for anti-macrophage monoclonal antibody CD68 (PGM1). Despite the compartmentalization of titanium within nodular areas in rats treated subcutaneously, systemic migration occurred. We concluded that systemic migration of TiO(2) occurred regardless of the administration route. PMID- 18389344 TI - Preparation of hydroxyapatite spheres with an internal cavity as a scaffold for hard tissue regeneration. AB - Microparticulates are currently regarded as a useful matrix for the delivery of bioactive molecules and tissue cells. Herein, hydroxyapatite (HA) spherical microparticulates with an internal cavity were produced using an oil-in-water emulsion technique. The HA slurry in the organic solvent was formulated into spherical particles in a water bath containing a surfactant. Rapid evaporation of the solvent helped create a cavity within the microparticulates. The microparticulates were heat-treated at 1,200 degrees C to produce bioactive HA particles with a mean size of approximately 363 microm. Osteoblastic cells were observed to spread and grow favorably over the surface as well as within the cavity of the microparticulates. The currently developed HA microparticulates with an internal cavity are considered to be useful as a scaffolding matrix for bone tissue engineering and direct filling skeletal defects. PMID- 18389345 TI - Dental casting alloys behaviour during power toothbrushing with toothpastes with various abrasivities. Part I: wear behavior. AB - The purpose of this study was to evaluate the long term effect of abrasivity of toothpastes normally used over the surface and mechanical properties of dental casting alloys. Three dental casting alloys (Ni-Cr, Co-Cr, c.p. Ti) and one ceramic were chosen. Four specimens of each material were immersed in artificial saliva, brushed without or with one of four toothpastes of different Relative Dentine Abrasivity (RDA 50, 52, 80, and 114). An electric toothbrush with a load of 250 g was used for 420 min. Mass loss was determined by difference in weight, microhardness and surface roughness were also measured. Two-way ANOVA and non parametric tests were used to detect significant differences. Titanium specimens (478 microg/cm(2)) exhibited the most mass loss, whereas ceramic (282 microg/cm(2)) and Co-Cr (262 microg/cm(2)) exhibited the least. However, ceramic demonstrated the most volume loss (0.239 mm(3)). The abrasivity effect of the toothpaste correlated with the RDA values. Slight variations in microhardness were observed after toothbrushing and depended on the material but not on the toothpaste used. Material surfaces were slightly smoothed by toothbrushing but no significant differences were detected. Dental casting alloys and ceramic are susceptible to abrasion by brushing with an electric toothbrush depending on the RDA value of the toothpaste. Variations in microhardness and surface roughness were not clinically relevant. PMID- 18389346 TI - Dental casting alloys behaviour during power toothbrushing with toothpastes of various abrasivities. Part II: corrosion and ion release. AB - The purpose of this study was to evaluate the long term effect of abrasivity of toothpastes normally used over the corrosion behavior and ion release of the different dental casting alloys. Three dental casting alloys (Ni-Cr, Co-Cr and commercially pure Ti) were studied. Four specimens of each material were immersed, brushed without paste or brushed with one of four toothpastes of different Relative Dentine Abrasivity (RDA 50, 52, 80, and 114). An electric toothbrush with a load of 250 g was used for 420 min. Corrosion behavior was determined by means a potenciostat with high sensitivity and the ion release determined by Inductively Coupled Plasma-Mass Spectrometry. Two-way ANOVA and non parametric tests were used to detect significant differences. Titanium specimens exhibited the best corrosion behavior after and before the toothbrushed, being the worst of the three alloys the Cr-Ni. Titanium oxide produced spontaneously on the Ti surface is the main cause of the high corrosion resistance of the material. However, the eutectoid of the CrNi with chemical composition between different phases produces pitting on the phases boundaries with an important decrease of the corrosion resistance. Besides, the CrNi produces high values of the Ni and Cr release. Slight increment in roughness were observed after toothbrushing and depended on the material but not on the toothpaste used. The increase of the microhardness (residual stresses) provokes a decrease of the corrosion resistance and an increase of the ion release. PMID- 18389347 TI - A novel amphiphilic nano hydrogel using ketene based polyester with polyacrylamide for controlled drug delivery system. AB - A ketene based Low molecular weight polymer (LMKP) having ester functional group was prepared using glycine through surface initiated anionic polymerization. NMR, ATR-FTIR & SEC were used to characterize the LMKP. The LMKP and acrylamide (AAm) were co-polymerised in methyl ethyl ketone to yield semi-IPN nanohydrogels (NHG). Benzoyl peroxide (BPO) was used as an initiator and N,N-methylene bisacrylamide (MBA) as crosslinking agent. Formation of NHG was confirmed through frequency shift in LMKP and poly acrylamide (PAAm) in FTIR spectroscopy. Photon correlation spectroscopy reveals that the sizes of the NHG were in the range of 140-225 nm and Transmission Electron Micrograph (TEM) also confirms the nano dimension of NHG. Biocompatibility of the NHG was confirmed through the cytotoxicity analysis. The swelling and diffusion behaviour of NHG, prepared under various formulations, were evaluated. The swelling pattern of NHG was studied at different pH conditions. The drug delivery capacity of NHG was investigated using ciprofloxacin as a model drug. The drug release kinetics of NHG suggested their anomalous (non-fickian) behaviour. PMID- 18389348 TI - Preparation and antibacterial characteristic of water-insoluble antibacterial material QPEI/SiO2. AB - Polyethyleneimine (PEI) was grafted onto micron-sized silica gel particles via the coupling action of gamma-chloropropyl trimethoxy silane, and grafting particles PEI/SiO(2) was prepared. Then, polymeric reactions of two steps, tertiary amination reaction and quaterisation, were conducted for the grafted PEI. After quaterisation of the grafted polyethyleneimine, a composite particle, QPEI/SiO(2), was obtained. QPEI/SiO(2) is a water-insoluble antibacterial material. In this work, the antibacterial characteristic of QPEI/SiO(2) was mainly investigated using Escherichia coli (E. coli) as a disease-leading bacterium and adopting a colony count method. The effects of quaterisation degree of PEI and pH of the medium on the antibacterial ability of QPEI/SiO(2) were examined. The antibacterial mechanism of QPEI/SiO(2) was explored profoundly by using two enzyme activity determination methods, beta-D-galactosidase activity determination and TTC-dehydrogenase activity determination. The experimental results indicates that the water-insoluble antibacterial material QPEI/SiO(2) possesses strong antibacterial ability, for the bacterial suspension with a concentration of 10(9) CFU/ml, the antibacterial ratio of QPEI/SiO(2) can reach about 100% only with a dosage of 15 g/l and only for a contact time of 10 min. The main factors influencing the antibacterial ratio of QPEI/SiO(2) are the quaterisation degree of the grafted PEI and pH value of the medium. QPEI/SiO(2) with higher quaterisation degree has stronger antibacterial ability. In a certain range of pH value, the higher the pH value of the medium is, the stronger the antibacterial ability of QPEI/SiO(2 )is. Enzyme activity determination results show that the antibacterial mechanism of the water-insoluble antibacterial material QPEI/SiO(2) is based on a sterilization process and not only is a bacteriostasis action. PMID- 18389349 TI - An uncertain dominion: Irish psychiatry, methadone, and the treatment of opiate abuse. AB - This paper investigates some productive ambiguities around the medical administration of methadone in the Republic of Ireland. The tensions surrounding methadone maintenance therapy (MMT) are outlined, as well as the sociohistorical context in which a serious heroin addiction problem in Ireland developed. Irish psychiatry intervened in this situation, during a time of institutional change, debates concerning the nature of addiction, moral panics concerning heroin addiction in Irish society and the recent boom in the Irish economy, known popularly as the Celtic Tiger. A particular history of this sort illuminates how technologies like MMT become cosmopolitan, settling into, while changing, local contexts. PMID- 18389350 TI - The medical exclusion of an immigrant to the United States of America in the early twentieth century. The case of Cristina Imparato. AB - The medical inspection of immigrants arriving in the United States in the late nineteenth and early twentieth centuries was framed by a need to rapidly process large numbers of people. Scientific medicine, such as it was then, was subordinate to the existing immigration laws which reflected the influences of the powerful anti-immigration forces of eugenics and nativism. The line, or single-file queues in which immigrants were arranged, facilitated rapid processing. The split-second medical gaze was hailed by the then leadership of the U.S. Public Health Service as scientifically sound, based as it was on the alleged exceptional disease detection skills of examining public health physicians. In reality, this system was seriously flawed, led to numerous diagnostic errors, and was free of any form of public outcomes assessment or accountability. In time, trachoma became the principal focus of line physicians because of the belief that it could be easily detected, and those diagnosed with it summarily deported. However, the diagnosis of the early stages of this disease is far more complex since it must be differentiated from other forms of benign conjunctivitis. Described here is the case of Cristina Imparato, a 46-year-old immigrant who arrived in New York from Italy on September 27, 1910, and who was given a diagnosis of trachoma which resulted in her summary deportation three days later on September 30, 1910. Her case serves to illustrate the complex forces at work at that time around the issue of immigration. These included a need to meet the labor needs of expanding industries while responding to the immigration restriction demands of eugenics supporters and nativists and the call to protect the country from imported disease threats. PMID- 18389351 TI - Factors influencing HIV vaccine community engagement in the urban South. AB - The purpose of this exploratory study was to examine personal characteristics, socio-environmental conditions, and motivational factors that potentially influence HIV vaccine research community engagement. Specifically, the study identified predictive aspects that may aid in future community program development on HIV vaccine issues. A cross-sectional survey consisting of evaluative measures, demographics, social interaction, and health information seeking behaviors was conducted. Participants were a diverse group of 452 adults (>or=18 years) at HIV vaccine awareness-building and community education gatherings in Atlanta. The sample included large numbers of women (n=251) and minorities (n=224). In multivariate analysis, the overall logistic regression model was significant, with a resulting coefficient of determination (Nagelkerke R(2)) of .505. Highly significant factors included an excellent activity/event rating (log odds beta = 4.521, P< .001), White race (beta= -.835, P= .005), greater educational attainment (beta= .725, P= .011), travel distance (beta = 1.186, P= .002), and excellent perception of the study site (beta=2.131, P< .001). Subgroup analyses by gender and race revealed similar findings. These data demonstrate the importance of building a favorable study site image and gaining familiarity in the community to aid in the promotion of HIV vaccine research on an ongoing basis. PMID- 18389352 TI - Three maternal risk factors associated with elevated risk of postneonatal mortality among Alaska native population. AB - OBJECTIVE: Compared to non-Natives in Alaska, the Alaska Native population has a postneonatal mortality rate 2.3 times higher (95% CI 1.9, 2.7). The objective of the study was to identify variables that account for this elevated risk. METHODS: The dataset used included birth and death certificate records for all Alaska resident live births and infant deaths occurring during 1992-2004. Race was defined as Alaska Native or non-Native. The association between race and postneonatal mortality was examined using univariate, stratified and regression analyses. Variables were considered confounding if they resulted in a change of at least 10% in the odds ratio between race and postneonatal mortality when added to a bivariate model, or when removed from a multivariate model. RESULTS: In stratified analysis, race remained associated with postneonatal mortality within most categories of marital status, maternal education, maternal age, prenatal tobacco or alcohol use, prenatal care utilization, parity and residence. The odds ratio between race and postneonatal mortality was reduced to 1.3 (95% CI 1.0, 1.6) by controlling for education, a composite variable of marital status and the presence of father's name on the birth certificate, and prenatal tobacco or alcohol use. CONCLUSIONS: A small number of potentially modifiable factors explain most of the postneonatal mortality disparity between Alaska Natives and non-Natives, leaving a relatively small increase in risk. These findings suggest that by targeting Alaska Native women who display these characteristics, the postneonatal mortality gap may be reduced. PMID- 18389353 TI - Evaluation of epicardial microwave lesions in the pig model using an electroanatomic mapping system. AB - BACKGROUND: Surgical ablation techniques using microwave energy are an alternative to catheter-based ablation therapy in the treatment of atrial fibrillation. We investigate electrical conduction properties of linear lesions after epicardial ablation in a pig model by using an electroanatomic mapping system. METHODS: After medial sternotomy in eight pigs epicardial pacing wires were placed on the anterior and posterolateral wall of the right atrium. Endocardial electroanatomical mapping was performed during pacing from anterior and posterolateral. Epicardial ablation was implemented in between the superior and inferior caval vein using microwave energy. Mapping was repeated to demonstrate conduction block or gap across the ablation line. When conduction gap was present the ablation procedure was repeated. Hearts were removed for histologic evaluation. RESULTS: Conduction block was observed in two pigs after the first ablation and in two additional pigs after the second ablation. Conduction gap was present in three pigs after the second ablation. One animal died during the second ablation. Histologic evaluation showed more severe transmural cellular damage in lesions with conduction gap. Complete lesions with conduction block more often showed moderate alterations. CONCLUSIONS: Epicardial microwave ablation can induce bidirectional conduction block, which is believed to be essential for therapeutic efficacy. However, the intraoperative macroscopic visible lesion and histological findings are not reliable markers of bidirectional conduction block. The Carto-Qwik-Map-System is an effective system to evaluate epicardial microwave lesions and to locate conduction gap areas intraoperatively. This increased the success rate regarding acute bidirectional conduction block after ablation in our experimental setting. PMID- 18389354 TI - Inner happiness among Thai elderly. AB - This study, based on data collected in 2005 from Chai Nat province, examines the level of happiness of the Thai elderly population and its relationship to various external and internal factors. It was found that mean happiness was slightly above a feeling of "neutral." According to multiple regression analyses, external factors including economic hardship, living arrangements, functional ability, perceived social environment, and consumerism significantly influence the level of happiness. The strongest predictor of happiness is, however, the internal factor-that is, a feeling of relative poverty when compared to their neighbors. Controlling for demographic and all external factors, the respondents who do not feel poor show the highest level of happiness compared to those who feel as poor as or poorer than their neighbors. This is self-interpreted as a feeling of contentment with what one has, which has been influenced by Thai culture, which is pervaded by Buddhism. PMID- 18389355 TI - A general test for gene-environment interaction in sib pair-based association analysis of quantitative traits. AB - Several association studies support the hypothesis that genetic variants can modify the influence of environmental factors on behavioral outcomes, i.e., G x E interaction. The case-control design used in these studies is powerful, but population stratification with respect to allele frequencies can give rise to false positive or false negative associations. Stratification with respect to the environmental factors can lead to false positives or false negatives with respect to environmental main effects and G x E interaction effects as well. Here we present a model based on Fulker et al. (1999) and Purcell (2002) for the study of G x E interaction in family-based association designs, in which the effects of stratification can be controlled. Simulations illustrate the power to detect genetic and environmental main effects, and G x E interaction effects for the sib pair design. The power to detect interaction was studied in eight different situations, both with and without the presence of population stratification, and for categorical and continuous environmental factors. Results show that the power to detect genetic and environmental main effects, and G x E interaction effects, depends on the allele frequencies and the distribution of the environmental moderator. Admixture effects of realistic effect size lead only to very small stratification effects in the G x E component, so impractically large numbers of sib pairs are required to detect such stratification. PMID- 18389356 TI - Knowledge, attitudes and motivations among blood donors in Sao Paulo, Brazil. AB - Recruiting safe, volunteer blood donors requires understanding motivations for donating and knowledge and attitudes about HIV. We surveyed 1,600 persons presenting for blood donation at a large blood bank in Sao Paulo, Brazil using a self-administered, structured questionnaire, and classified motivations into three domains as well as categorizing persons by HIV test-seeking behavior. Motivations, in descending order, and their significant associations were: "altruism": female gender, volunteer donor and repeat donor status; "direct appeal": female gender, repeat donor status and age 21-50 years; "self-interest": male gender, age under 20 years, first-time donor status and lower education. HIV test-seekers were more likely to give incorrect answers regarding HIV risk behavior and blood donation and the ability of antibody testing to detect recent HIV infections. Altruism is the main motivator for blood donation in Brazil; other motivators were associated with specific demographic subgroups. HIV test seeking might be reduced by educational interventions. PMID- 18389357 TI - An empirical comparison of respondent-driven sampling, time location sampling, and snowball sampling for behavioral surveillance in men who have sex with men, Fortaleza, Brazil. AB - Obtaining samples of populations at risk for HIV challenges surveillance, prevention planning, and evaluation. Methods used include snowball sampling, time location sampling (TLS), and respondent-driven sampling (RDS). Few studies have made side-by-side comparisons to assess their relative advantages. We compared snowball, TLS, and RDS surveys of men who have sex with men (MSM) in Forteleza, Brazil, with a focus on the socio-economic status (SES) and risk behaviors of the samples to each other, to known AIDS cases and to the general population. RDS produced a sample with wider inclusion of lower SES than snowball sampling or TLS a finding of health significance given the majority of AIDS cases reported among MSM in the state were low SES. RDS also achieved the sample size faster and at lower cost. For reasons of inclusion and cost-efficiency, RDS is the sampling methodology of choice for HIV surveillance of MSM in Fortaleza. PMID- 18389358 TI - Accessing men who have sex with men through long-chain referral recruitment, Guangzhou, China. AB - Men who have sex with men (MSM) may account for an increasing proportion of China's HIV epidemic, but remain difficult to access for epidemiological studies due to high stigma. We compare the composition of two samples of MSM obtained in Guangzhou, China. The first survey, conducted in 2004, recruited MSM through convenience sampling. The second survey in 2006 used long-chain referral recruitment in the context of respondent-driven sampling. Compared to convenience sampling, the long-chain referral method included higher proportions of subgroups of MSM thought to be at elevated risk for HIV infection and more difficult to reach, including internal migrants and those engaging in commercial sex. Long chain referral also recruited more MSM who were under 25 years, unemployed, and had lower education. We conclude that long-chain referral recruitment will be more effective in tracking the leading edge of the epidemic among MSM in China than convenience sampling. PMID- 18389359 TI - Increasing HIV prevalence among drug users in western Guangdong province, China, 1995-2005. AB - Sentinel surveillance data from 1995 to 2005 for drug users in Guangdong province, China, showed an increasing prevalence of HIV in the West region while stabilizing in the East and Center. Several factors were significantly associated with HIV infection including gender, age, sharing needles, years injecting, engaging in commercial sex, and being part of the migrant population of Guangdong. Data help effectively prioritize and target HIV prevention efforts for drug users. PMID- 18389360 TI - Between tradition and change: condom use with primary sexual partners among Mexican migrants. AB - The frequency of male Mexico-US migration has been associated with increased HIV risk for sexual partners awaiting their return in Mexico. This study examined the association between sexual partner characteristics and condom use among a sample of 354 male migrants from two Mexican municipalities. Migrants were interviewed about their past year's sex practices. Results indicated that migrants were more likely to use condoms with their non-spousal partners, partners with less education than the migrant, and partners with higher employment status. Condom use was greater among younger migrants and residents of the more densely populated municipality. Findings suggest the coexistence of a traditional cultural orientation that does not support condom use and another one that does provided the sex partner is formally employed. Prevention programs must strengthen the structural conditions fostering greater equality between the sexes and adapt their approaches for different population density, age and partner types. PMID- 18389361 TI - Patterns and correlates of sexual activity and condom use behavior in persons 50 plus years of age living with HIV/AIDS. AB - This study characterized rates of sexual activity and identified psychosocial and behavioral correlates of sexual activity and condom use in a metropolitan sample of 290 HIV-infected adults 50-plus years of age. Thirty-eight percent of participants were sexually active in the past three months, 33% of whom had at least one occasion of anal or vaginal intercourse that was not condom protected. Rates and correlates of sexual activity and condom use differed between gay/bisexual men, heterosexual men, and heterosexual women. In the past three months, 72% of heterosexual men were sexually active compared to only 36% of gay/bisexual men and 21% of heterosexual women. However, among sexually active persons, only 27% of heterosexual men reported inconsistent condom use compared to 37% of gay/bisexual men and 35% of heterosexual women. As the number of older adults living with HIV/AIDS in the U.S. continues to increase, age-appropriate secondary risk-reduction interventions are urgently needed. PMID- 18389362 TI - Starting young: sexual initiation and HIV prevention in early adolescence. AB - The rising numbers of new HIV infections among young people ages 15-24 in many developing countries, especially among young women, signal an urgent need to identify and respond programmatically to behaviors and situations that contribute to the spread of HIV and other sexually transmitted infections in early adolescence. Quantitative and qualitative studies of the sexual knowledge and practices of adolescents age 14 and younger reveal that substantial numbers of boys and girls in many countries engage in unprotected heterosexual vaginal intercourse--by choice or coercion--before their 15th birthdays. Early initiation into male-male or male-female oral and/or anal sex is also documented in some populations. Educational, health, and social programs must reach 10-14-year-olds as well as older adolescents with the information, skills, services, and supplies (condoms, contraceptives) they need to negotiate their own protection from unwanted and/or unsafe sexual practices and to respect the rights of others. PMID- 18389363 TI - Development and psychometric assessment of a multidimensional measure of internalized HIV stigma in a sample of HIV-positive adults. AB - There is a need for a psychometrically sound measure of the stigma experienced by diverse persons living with HIV/AIDS (PLHA). The goal of this study was to develop and evaluate a multidimentional measure of internalized HIV stigma that captures stigma related to treatment and other aspects of the disease among sociodemographically diverse PLHA. We developed a 28-item measure of internalized HIV stigma composed of four scales based on previous qualitative work. Internal consistency reliability estimates in a sample of 202 PLHA was 0.93 for the overall measure, and exceeded 0.85 for three of the four stigma scales. Items discriminated well across scales, and correlations of the scales with shame, social support, and mental health supported construct validity. This measure should prove useful to investigators examining in the role of stigma in HIV treatment and health outcomes, and evaluating interventions designed to mitigate the impacts of stigma on PLHA. PMID- 18389364 TI - Antiretroviral therapy is associated with increased fertility desire, but not pregnancy or live birth, among HIV+ women in an early HIV treatment program in rural Uganda. AB - To assess the association between antiretroviral therapy (ART) and fertility history and desire among HIV-positive Ugandan women, we conducted a cross sectional study among HIV-positive Ugandan women aged 18-50 years who attended an HIV clinic at Mbarara University in western Uganda between November 1, 2005 and June 6, 2006. Of 538 women approached, 501 were enrolled. ART use was associated with increased odds of fertility desire (AOR 2.99, 95% CI 1.38-6.28), and decreased odds of pregnancy (AOR 0.56, 95% CI 0.33-0.95) and live birth (AOR 0.30, 95% CI 0.13-0.66). ART was associated with an increase in fertility desire, but was not associated with an increase in fertility. Additional studies will be needed to determine if this greater fertility desire among ART-treated women leads to an increase in fertility as ART use expands. PMID- 18389365 TI - Pulsed reduced dose-rate radiotherapy: a novel locoregional retreatment strategy for breast cancer recurrence in the previously irradiated chest wall, axilla, or supraclavicular region. AB - PURPOSE: Reirradiation of breast cancer locoregional recurrence (LRR) in the setting of prior post-mastectomy radiation poses a significant clinical challenge due to the high risk for severe toxicity. In an attempt to reduce these toxicities, we have developed pulsed reduced dose-rate radiotherapy (PRDR), a reirradiation technique in which a series of 0.2 Gy pulses separated by 3-min time intervals is delivered, creating an apparent dose rate of 0.0667 Gy/min. Here we describe our early experience with PRDR. PATIENTS AND METHODS: We reirradiated 17 patients with LRR breast cancer to the chest wall, axilla, or supraclavicular region using PRDR. The median prior radiation dose was 60 Gy. We delivered a median PRDR dose of 54 Gy (range 40-66 Gy) in 1.8-2.0 Gy per fraction. Eight patients received concomitant low dose capecitabine for radiosensitization. The median treatment volume was 2,084 cm(3) (range 843-7,881 cm(3)). RESULTS: At a median follow-up of 18 months (range 4-75 months) only 2 patients have had tumor failure in the treatment region. Estimated 2-year local control rate is 92%. Treatment was well tolerated with 4 patients experiencing grade 3 acute skin toxicity. Despite a median cumulative dose of 110 Gy (range 80 236 Gy), there has been only one grade 3 and one grade 4 late toxicity. CONCLUSIONS: With a median follow-up of 18 months, PRDR appears to be an effective method to reirradiate large volumes of previously irradiated tissue in selected patients with locoregional chest wall, axilla, and supraclavicular recurrences. PMID- 18389366 TI - Axillary recurrence after negative sentinel lymph node biopsy. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) has almost totally replaced axillary lymph node dissection as the first-line axillary procedure for node negative breast cancer. SLNB has a false-negative rate of 0-22%, and regional nodal recurrence is a major concern after SLNB. In this study, we assessed axillary recurrence and risk factors in breast cancer patients 40 months after negative SLNB. METHODS: Of 940 patients with node-negative breast cancer who underwent SLNB between December 2003 and January 2006 at Asan Medical Center, 720 were negative on SLNB, as determined using 99-m TC radiocolloid and subareolar injection technique. Of the 720 patients negative on SLNB, 174 underwent further axillary dissection, 253 underwent node sampling, and 293 received SLNB only. RESULTS: A mean of 2.1 SLNs was removed per patient. At a median follow-up of 40 months (range 24-49 months), recurrence in the axilla was observed in three patients, all of whom had undergone SLNB only; two of these patients also had recurrences in internal mammary lymph nodes. Tumors in all three patients were hormone-receptor negative, and two were c-erbb2 negative. CONCLUSION: The axillary recurrence rate was low in patients negative on SLNB. Negative hormone receptor status and high nuclear grade may be risk factors for regional nodal failure after SLNB. PMID- 18389367 TI - Lifestyle changes and breast cancer prognosis: a review. AB - OBJECTIVE: Because of its high incidence and improving survival, breast cancer is currently the most prevalent cancer in the world. We reviewed the published literature in our search for modifiable factors valuable as an adjuvant health measure to surgery, radio and systemic therapy for breast cancer. METHODS: We included material which was identified from computerised searches of PubMed (1966 to May, 2007). Published material was restricted to prospective cohort studies and randomised clinical trials. We reviewed the literature concerning the association between physical activity, smoking, weight gain after the diagnosis, diet and prognosis. RESULTS AND CONCLUSION: We conclude that life style changes following standard breast cancer therapy are highly recommended (although scientific evidence is still lacking for some of inconsistency regarding available data). They include physical activity, weight control, high consumption of fruit and vegetables, and a reduction of dietary fat intake. PMID- 18389368 TI - Predictors of follow-up exercise behavior 6 months after a randomized trial of exercise training during breast cancer chemotherapy. AB - PURPOSE: Exercise during breast cancer chemotherapy is beneficial but it needs to be maintained into survivorship to optimize long-term benefits. Here, we report the predictors of follow-up exercise behavior 6 months after a randomized exercise trial in breast cancer patients. METHODS: Breast cancer patients (N = 242) initiating adjuvant chemotherapy were randomly assigned to usual care (n = 82), supervised resistance exercise (n = 82), or supervised aerobic exercise (n = 78) for the duration of their chemotherapy. At baseline and postintervention, data were collected on demographic, medical, behavioral, fitness, psychosocial, and motivational variables. At 6-month follow-up, participants were mailed a questionnaire that assessed exercise behavior over the past 6 months and were categorized as either meeting both aerobic and resistance exercise guidelines, either exercise guideline, or neither exercise guideline. RESULTS: Two hundred one (83.1%) participants provided 6-month follow-up data with 85 (42.3%) meeting neither exercise guideline, 74 (36.8%) meeting either exercise guideline, and 42 (20.9%) meeting both exercise guidelines. In multivariate regression analysis, seven variables independently predicted the likelihood of meeting exercise guidelines at follow-up including higher pretrial exercise (beta = 0.23; P = 0.002), younger age (beta = -0.15; P = 0.028), breast conserving surgery (beta = 0.15; P = 0.033), strength improvements (beta = 0.15; P = 0.028), lower postintervention fatigue (beta = 0.13; P = 0.067), a more positive attitude (beta = 0.12; P = 0.086), and lower postintervention body mass index (beta = -0.11; P = 0.105). CONCLUSION: Exercise behavior 6 months after a randomized trial was predicted by a wide range of demographic, medical, behavioral, fitness, psychosocial, and motivational variables. These findings may help facilitate the uptake of exercise behavior during the transition from breast cancer patient to survivor. PMID- 18389369 TI - Polysaccharides as a protective response against microcystin-induced oxidative stress in Chlorella vulgaris and Scenedesmus quadricauda and their possible significance in the aquatic ecosystem. AB - Toxic cyanobacteria occur worldwide in aquatic ecosystem, and their toxins have adverse effects on most aquatic organisms. However, some species of green algae can grow and flourish at environmentally relevant concentrations of microcystins (MCYSTs). Therefore, the present study aimed to investigate the possible adaptive response of two representatives of green algae, Chlorella vulgaris and Scenedesmus quadricauda to these toxins. Growth and antioxidative biomarkers of these algae were studied over a 14-day exposure to different concentrations of pure microcystin-LR (MCYST-LR) and crude MCYSTs. Both pure and crude MCYSTs significantly decreased the growth of the two algae compared to control cultures during the first 3 days of incubation. Meanwhile, increases in glutathione-S transferase (GST), glutathione peroxidase (GPX) and lipid peroxidation, and decreases in glutathione (GSH) were also observed in toxin-treated cultures. All growth and biochemical variables were restored to control levels after 3 days of incubation and remained at levels near to those of control cultures during the remaining period of experiment. The changes in these variables correlated with polysaccharide contents of toxin-treated cultures, indicating the involvement of these polysaccharides in protecting the algal cells against MCYST-induced oxidative stress. The results of in vitro assay of antioxidant activity revealed that these polysaccharides had different activities, depending on their sulfate contents. This study provides an evidence for the first time that polysaccharides play a protective role in some microalgae against MCYST-induced oxidative stress. PMID- 18389370 TI - Clapper rails as indicators of mercury and PCB bioavailability in a Georgia saltmarsh system. AB - Clapper rails (Rallus longirostris) were used as an indicator species of estuarine marsh habitat quality because of their strong site fidelity and predictable diet consisting of mostly benthic organisms. Mercury (Hg) and the polychlorinated biphenyl (PCB) Aroclor 1268 concentrations were determined for sediments, crabs, as well as clapper rail adults and chicks collected from salt marshes associated with the LCP Superfund site in Brunswick, Georgia. Home ranges were established for adult rails, and sediment and crab samples were taken from each individual's range. The study was designed to minimize the spatial variability associated with trophic transfer studies by choosing an endpoint species with a potentially small home range and specifically sampling its foraging range. The mean home range for clapper rails was 1.2 ha with a median of 0.28 ha. Concentrations of Hg and Aroclor 1268 were shown to increase with each trophic level. Transfer factors between media followed the same pattern for both contaminants with the highest between fiddler crabs and clapper rail liver. Hg and PCB transfer factors were similar between sediment to fiddler crab and fiddler crab to muscle, however the PCB transfer factor from fiddler crabs to liver was over twice as large as for Hg. PCB congener profiles did not significantly differ between media types. PMID- 18389371 TI - Oxidative status of Matricaria chamomilla plants related to cadmium and copper uptake. AB - Cadmium (Cd) and copper (Cu) uptake by the plants of Matricaria chamomilla and relation to activities of guaiacol peroxidase (GPX, EC 1.11.1.7), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) up to 7 days of exposure to 3, 60 and 120 microM Cd or Cu was studied. Cd content in rosettes was ca. 10 fold higher in comparison to Cu while Cu was preferentially accumulated in the roots. In line with this observation, increase of CAT and GPX activity was similar in rosettes of Cd and Cu-treated plants, indicating non-redox active properties of Cd and low Cu accumulation. In the roots, Cu showed strong pro oxidant effect, as judged from extreme stimulation of CAT and GPX, followed by increase of hydrogen peroxide and malondialdehyde. However, GPX seemed to be more important for alleviation of oxidative stress (ca. 93-250-fold higher activity in 120 microM Cu-treated roots). Cd had substantially lower influences and stimulated GR activity more than Cu. Activities of hydrogen peroxide-scavenging enzymes in relation to its accumulation are also discussed. PMID- 18389372 TI - Clenching TMJs-loads increases in partial edentates: a 3D finite element study. AB - GOAL: This study tests the hypothesis of loading-dependence on the temporomandibular joint during clenching on the particular of experimentally partial edentate conditions. METHODOLOGY: A complete and detailed finite element model of the temporomandibular joint (TMJ) was used. The closing movement of the mouth was reproduced by contracting the closing muscles of the masticatory system. Electromyography (EMG) data were taken from 10 healthy, dentulate volunteers, both with and without intraoral appliances. The intraoral appliances served to mimic nine partially edentulate (PE) conditions for each volunteer. The EMG data were fed into the finite element model (FEM) for each condition and the loading of the joint was analyzed. RESULTS: The results obtained show that muscular activity decreases when the contact between teeth disappears. In particular, the numerical results showed that when there is no contact between the posterior teeth an overload of the joints appeared. Moreover, the existence of a unilateral unique molar induced asymmetric overloading in the TMJ disc without posterior contact. CONCLUSIONS: During clenching, a uniform distribution of the dental contact along the maxillar arches prevents the TMJ from overloading. In contrast, severe partial edentation seems to induce overloading of the TMJ with severity depending on the type of contact. PMID- 18389374 TI - Reducing youth exposure to alcohol ads: targeting public transit. AB - Underage drinking is a major public health problem. Youth drink more heavily than adults and are more vulnerable to the adverse effects of alcohol. Previous research has demonstrated the connection between alcohol advertising and underage drinking. Restricting outdoor advertising in general and transit ads in particular, represents an important opportunity to reduce youth exposure. To address this problem, the Marin Institute, an alcohol industry watchdog group in Northern California, conducted a survey of alcohol ads on San Francisco bus shelters. The survey received sufficient media attention to lead the billboard company, CBS Outdoor, into taking down the ads. Marin Institute also surveyed the 25 largest transit agencies; results showed that 75 percent of responding agencies currently have policies that ban alcohol advertising. However, as the experience in San Francisco demonstrated, having a policy on paper does not necessarily mean it is being followed. Communities must be diligent in holding accountable government officials, the alcohol industry, and the media companies through which advertising occurs. PMID- 18389373 TI - Radiotherapy of other sellar lesions. AB - Radiotherapy has been used as primary or adjuvant treatment in a number of non adenomatous masses arising from within the sella. It is particularly important in the therapeutic algorithm of craniopharyngiomas and meningiomas and has also been used in chordomas/chordosarcomas and less commonly, in other lesions. This review describes the place of irradiation in the management of these masses. PMID- 18389375 TI - Australia's new urbanists encourage walking. PMID- 18389376 TI - Provision and use of maternal health services among urban poor women in Kenya: what do we know and what can we do? AB - In sub-Saharan Africa, the unprecedented population growth that started in the second half of the twentieth century has evolved into unparalleled urbanization and an increasing proportion of urban dwellers living in slums and shanty towns, making it imperative to pay greater attention to the health problems of the urban poor. In particular, urgent efforts need to focus on maternal health. Despite the lack of reliable trend data on maternal mortality, some investigators now believe that progress in maternal health has been very slow in sub-Saharan Africa. This study uses a unique combination of health facility- and individual-level data collected in the slums of Nairobi, Kenya to: (1) describe the provision of obstetric care in the Nairobi informal settlements; (2) describe the patterns of antenatal and delivery care, notably in terms of timing, frequency, and quality of care; and (3) draw policy implications aimed at improving maternal health among the rapidly growing urban poor populations. It shows that the study area is deprived of public health services, a finding which supports the view that low income urban residents in developing countries face significant obstacles in accessing health care. This study also shows that despite the high prevalence of antenatal care (ANC), the proportion of women who made the recommended number of visits or who initiated the visit in the first trimester of pregnancy remains low compared to Nairobi as a whole and, more importantly, compared to rural populations. Bivariate analyses show that household wealth, education, parity, and place of residence were closely associated with frequency and timing of ANC and with place of delivery. Finally, there is a strong linkage between use of antenatal care and place of delivery. The findings of this study call for urgent attention by Kenya's Ministry of Health and local authorities to the void of quality health services in poor urban communities and the need to provide focused and sustained health education geared towards promoting use of obstetric services. PMID- 18389377 TI - The sweet potato sporamin promoter confers high-level phytase expression and improves organic phosphorus acquisition and tuber yield of transgenic potato. AB - The sweet potato sporamin promoter was used to control the expression in transgenic potato of the E. coli appA gene, which encodes a bifunctional enzyme exhibiting both acid phosphatase and phytase activities. The sporamin promoter was highly active in leaves, stems and different size tubers of transgenic potato, with levels of phytase expression ranging from 3.8 to 7.4% of total soluble proteins. Phytase expression levels in transgenic potato tubers were stable over several cycles of propagation. Field tests showed that tuber size, number and yield increased in transgenic potato. Improved phosphorus (P) acquisition when phytate was provided as a sole P source and enhanced microtuber formation in cultured transgenic potato seedlings when phytate was provided as an additional P source were observed, which may account for the increase in leaf chloroplast accumulation (important for photosynthesis) and tuber yield of field grown transgenic potato supplemented with organic fertilizers. Animal feeding tests indicated that the potato-produced phytase supplement was as effective as a commercially available microbial phytase in increasing the availability of phytate-P to weanling pigs. This study demonstrates that the sporamin promoter can effectively direct high-level recombinant protein expression in potato tubers. Moreover, overexpression of phytase in transgenic potato not only offers an ideal feed additive for improving phytate-P digestibility in monogastric animals but also improves tuber yield, enhances P acquisition from organic fertilizers, and has a potential for phytoremediation. PMID- 18389379 TI - Benthic foraminifera from polluted marine environment of Sulaibikhat Bay (Kuwait). AB - Quantitative analyses of recent benthic foraminiferal assemblages (living and dead) were carried out on the surface sediments of Sulaibikhat Bay. Marked contrast in foraminiferal assemblages between the shallow tidal mudflats and the deep tidal channel and their relation to the extent of pollution were observed. Cluster analysis of quantitative data on the distribution of foraminiferal tests revealed three assemblages that depend mainly on the intensity of pollution; (1) a highly polluted tidal flat assemblage, (2) normal (or less polluted) mud flat assemblage and, (3) tidal channel and subtidal assemblage. The highly polluted assemblage characterized by a drop in species densities (< 100 tests/20 cm(3) sediment) but with high average diversity (5.8 Yule-Simpson Index). The microfauna of the less polluted flat displays relatively lower diversity (4.6) but high density of tests (47.2% of the total picked tests). The most abundant species of this assemblage is Ammonia tepida, displays its maximum density in this assemblage. Ammonia tepida drops in density from 17.12% to 3.07% in the polluted assemblage. Tidal channel foraminiferal assemblages should normally display lower diversities than those of tidal flats, because tidal current in the channels tend to wash away most nutrient materials. However, this is not the case of the present study area which could be due to environmental setting of the Sulaibikhat Bay in which tidal currents bring in exceptionally high amounts of nutrients from Shatt Al-Arab Estuary and in which the tidal flats are strongly and adversely polluted. PMID- 18389380 TI - RNAi-mediated knock-down of gene mN6A1 reduces cell proliferation and decreases protein translation. AB - Methyltransferases play essential roles in modulating important cellular and metabolic processes. A mouse putative N6-DNA methyltransferase gene (GenBank No AY456393) is a novel gene named mN6amt1(mN6A1). To investigate its function in cell fate and protein translation, RNA interference (RNAi)-mediated knock-down method was established. Cell cycle analysis suggests that the cell proliferation decreases after RNAi with mN6A1. The expression plasmid of luciferase was used to detect protein translation, and the results showed that luciferase expression decreased after RNAi with mN6A1, whereas increased after over-expression of mN6A1 or/and eRF1. The binding between mN6A1 and eRF1 was identified by co immunoprecipitation and pull-down experiments. It might be suggested that mN6A1 participates in protein translation through interaction with eRF1. PMID- 18389378 TI - A prospective study of educational background and breast cancer among Japanese women. AB - OBJECTIVE: This prospective cohort study examined the association between educational level and breast cancer incidence in Japan. METHOD: A baseline survey was conducted between 1988 and 1990 among 110,792 residents of 45 areas, aged 40 79 years. Data were restricted to 24 areas where incidence registry data were available, and to subjects which provided information on educational level (32,646). The subjects were assigned to three groups according to their level of education (<16, 16-18, 18<). During 13 years of follow-up (328,931 person-year), 169 cases of breast cancer were newly diagnosed. RESULTS: Women with a high level of education had an increased risk of breast cancer (HR = 1.93, 95 percent confidence interval (95% CI): 1.18, 3.16, in women with the highest educational level) compared with women with the lowest educational level. Adjustment for lifestyle and reproductive factors did not substantially change the results. In addition, when analyses were stratified by age subgroups, the educational difference in breast cancer incidence was more evident among the younger than the elder subgroup. CONCLUSION: The present results suggested that cancer prevention strategies should recognize women with a higher educational level as a high risk group for breast cancer. PMID- 18389381 TI - Protective effects of N-acetylcysteine in isoproterenol-induced myocardium injury in rats. AB - Isoproterenol (ISO) has been found to cause severe injury in the myocardium. The aim of this study was to investigate the protective effects of N-acetylcysteine (NAC) on ISO-induced myocardial injury in rats and its underlying mechanisms. Fouty male Wistar rats were randomly divided into four groups: control, ISO, NAC, and ISO + NAC group. Myocardial histopathological observation were performed; The activities of creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) were examined; Myocardium TNF-alphaand IL-1beta gene expressions were examined by RT-PCR analysis; Myocardial expressions of TNF-alphaand IL 1betaproteins were observed by immunohistochemical assay and western blotting analysis. The myocardial injury induced by ISO was significantly reduced by the treatment of NAC as judged by the reduction of myocardial necrosis. Compared with ISO group, rats pre-injected with NAC showed a significant decrease in the activities of cardiac marker enzymes such as CK-MB and LDH in serum. NAC inhibits the pro-inflammatory factors expressions (TNF-alphaand IL-1beta) stimulated by ISO. In conclusion, NAC exerts significant cardio-protective effects against ISO induced myocardial injury in rats, likely regulating pro-inflammatory factors expressions. PMID- 18389382 TI - The spectrum of FMF mutations and genotypes in the referrals to molecular genetic laboratory at Kirikkale University in Turkey. AB - Familial Mediterranean Fever (FMF) is an autosomal recessive genetic disorder characterised by recurrent and self-limited abdominal pain, synovitis and pleuritis. MEFV gene mutations are responsible from the disease and its protein product, pyrin or marenostrin, plays an essential role in the regulation of the inflammatory reactions. MEFV gene contains 10 exons and most of the mutations have been found on the last exon. Up to date, 152 mutations and polymorpisms have been reported inwhere V726A, M694V, M694I, M680I and E148Q are the most common mutations. In this study, MEFV allele frequencies of 136 individuals (60 from Pediatry, 76 from Internal Medicine) have been evaluated, and compared with each other. Asymptomatic individuals with FMF family history (4 from Pediatry, 6 from Internal Medicine) were excluded from the analysis. The prominent mutations indicated in the Pediatry group are V726A, M694V and M680I (G/C) and with the allele frequency of 0.06, 0.05 and 0.04 respectively while they were E148Q, M694V, M680I (G/C) in the Internal Medicine group with the allele frequency of 0.12, 0.08 and 0.04. The E148Q mutation is significantly overrepresented in the adult referrals (P = 0.02). Mutation on both alleles was observed in only 12% of cases. Overall mutation frequency was low, seen in 26.2% (66/252). However, when only diagnosed patients were analyzed it is 72.7% (16/22). It is also interesting that 63% of individuals are female that there may be sex influence on FMF phenotype. PMID- 18389383 TI - Reliability and validity of childhood asthma control test in a population of Chinese asthmatic children. AB - BACKGROUND: No quick and easy assessment tool is presently available to measure asthma control and outcome in Chinese children. METHODS: We translated an established seven-item questionnaire into the Chinese Childhood Asthma Control Test (C-CACT) to measure asthma symptoms and their effect on daily function in 247 children aged 4-11 years enrolled from specialty clinics around Tainan. Subjects were classed as having either intermittent or persistent asthma and were evaluated at two outpatient visits 4-8 weeks apart (baseline and follow-up). RESULTS: C-CACT at baseline correlated significantly with physician evaluation scores (PES) except in predicted peak expiratory flow rate. There were significant differences for mean C-CACT score and mean PES between the intermittent group and persistent group. Internal consistency reliability of the survey was 0.741 at baseline and 0.759 at the follow-up visit. Test-retest reliability among 209 patients with the same specialist rating of asthma control at baseline and follow-up was r = 0.546 (P < 0.001). C-CACT scores of patients whose therapy needed to be stepped up was significantly lower than in those with unchanged or stepped-down therapy (F = 39.61, P < 0.001). CONCLUSIONS: C-CACT provides a reliable, valid, and responsive measure of asthma control in Chinese children. PMID- 18389385 TI - The factorial structure of the Chronic Liver Disease Questionnaire (CLDQ). AB - OBJECTIVE: The Chronic Liver Disease Questionnaire (CLDQ) is a disease-specific instrument designed to assess health-related quality of life in patients with chronic liver disease. The aim of this paper is to present the psychometric properties of a German version of this questionnaire. A special focus is placed on the analysis of the CLDQ's factorial structure. METHODS: Five hundred and twenty-four patients completed the CLDQ from May 1999 to October 2006. The results were subject to item analysis, reliability and validity assessments, and confirmatory and exploratory factor analysis. RESULTS: The distribution characteristics on the item and scale level were satisfactory. Internal consistency was good to excellent; retest reliability acceptable. Validity could be confirmed by characteristic subscale correlations with other quality-of-life scales. Confirmatory factor analysis could not sufficiently reproduce the original factor structure. Exploratory factor analysis suggested five out of six subscales of the original version and yielded a new subscale: leep." CONCLUSION: CLDQ's reliability and validity have been confirmed. In addition, the demonstrated practical administration of the questionnaire suggests that it should serve as a routine quality of life assessment of patients with chronic liver disease. PMID- 18389384 TI - Listen to their answers! Response behaviour in the measurement of physical and role functioning. AB - BACKGROUND: Quality of life (QoL) is considered to be an indispensable outcome measure of curative and palliative treatment. However, QoL research often yields findings that raise questions about what QoL measurement instruments actually assess and how the scores should be interpreted. OBJECTIVE: To investigate how patients interpret and respond to questions on the EORTC-QLQ-C30 over time and to find explanations to account for counterintuitive findings in QoL measurement. METHODS: Qualitative investigation was made of the response behaviour of small cell lung cancer patients (n = 23) in the measurement of QoL with the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC QLQ-C30). Focus was on physical functioning (PF, items 1 to 5), role functioning (RF, items 6 and 7), global health and QoL rating (GH/QOL, items 29 and 30). Interviews were held at four points: at the start of the chemotherapy, 4 weeks later, at the end, and 6 weeks after the end of chemotherapy. Patients were asked to 'think aloud' when filling in the questionnaire. RESULTS: Patients used various response strategies when answering questions about problems and limitations in functioning, which impacted the accuracy of the scale. Patients had scores suggesting they were less limited than they actually were by taking the wording of questions literally, by guessing their functioning in activities that they did not perform, and by ignoring or excluding certain activities that they could not perform. CONCLUSION: Terminally ill patients evaluate their functioning in terms of what they perceive to be normal under the circumstances. Their answers can be interpreted in terms of change in the appraisal process (Rapkin and Schwartz 2004; Health and Quality of Life Outcomes, 2, 14). More care should be taken in assessing the quality of a set of questions about physical and role functioning. PMID- 18389386 TI - Screening for urinary tract cancer with urine cytology in Lynch syndrome and familial colorectal cancer. AB - AIM: The aim of this study was to evaluate if Urine Cytology (UC) is an appropriate screening procedure for detecting urinary tract neoplasia at an early stage in persons at risk in Hereditary Non-Polyposis Colorectal Cancer families. METHOD: In the National Danish HNPCC-register persons at risk were identified in three categories of HNPCC-families (1) families harbouring a disease causing mutation in a Mismatch repair gene (MMR), (2) families fulfilling the Amsterdam I or II criteria and (3) families suspected of HNPCC. In total 3,411 persons were identified and traced in Patobank-the National Danish Pathology database. All UC and UTC (Urinary Tract Tumours) were listed and evaluated. RESULTS: 977 persons had a total of 1,868 screening procedures performed. Two of these procedures (0.1%) lead to diagnosis of an asymptomatic urothelial tumour. In ten times as many procedures (22 persons) UC lead to a false positive screening diagnosis. During the study period fourteen persons (1.4%) developed a UTC and five of these were interval tumours. The sensitivity of UC in diagnosing asymptomatic UTC in HNPCC patients was 29%. Twelve of the tumours were found in persons from families with a proven MMR-mutation and eleven out of these were MSH2 mutations (92%, 95% cl 62-100%). DISCUSSION: UC is not a proper method of screening for UTC in HNPCC. However, the study can not reveal if screening for UTC in special families ought to be recommended. Consequently, further studies needs to be performed in order to evaluate an appropriate screening programme. PMID- 18389387 TI - Cascade genetic testing for mismatch repair gene mutations. AB - Mismatch repair gene mutation carriers have a high risk of developing colorectal cancer, and can benefit from appropriate surveillance. A combined population based ascertainment cascade genetic testing approach provides a systematic and potentially effective strategy for identifying such carriers. We have developed a Markov Chain computer model system which simulates various factors influencing cascade genetic testing; including demographics, uptake, genetic epidemiology and family size. This was used to evaluate cascade genetic testing for mismatch repair gene mutations in theory and practice. Simulations focussed on the population of Scotland by way of illustration, and were based on a 20-year programme in which index cases were ascertained from colorectal cancer cases aged<55 years at onset. Results indicated that without practical barriers to cascade genetic testing, 545 (95% CI=522, 568) carriers could be identified; 42% of the population total. This comprised approximately 140 index cases, 302 asymptomatic relatives and 104 previously affected relatives. However, when realistic ascertainment and acceptance rates were used to inform simulations, only 257 (95% CI=246, 268) carriers, about 20% of the carrier population, were identifiable. Of these approximately 112 were index cases, 108 were asymptomatic relatives, and 37 were previously affected relatives. This contrast emphasises the importance of ascertainment and acceptance rates. Likewise the low number of index cases shows that case identification is a limiting factor. In the absence of robust data from epidemiological studies, these findings can inform decisions about the use of cascade genetic testing for mismatch repair gene mutations. PMID- 18389388 TI - Mutation spectrum in HNPCC in the Israeli population. AB - Hereditary non-polyposis colon cancer is caused by mutations in DNA mismatch repair genes. The mutation spectrum in the Israeli population is poorly documented except for the c.1906G>C Ashkenazi founder mutation in the hMSH2 gene. To report our experience in HNPCC screening, the mutations detected and the clinical features among a cohort of Israeli patients. Diagnostic work-up was done in a multi-step process guided by clinical and ethnic information. Tumors of suspected patients were tested for microsatellite instability and immunohistochemistry. Based on tumor analyses, we proceeded to mutation screening by DHPLC followed by sequence analysis and multiplex ligase dependent probe amplification. Ashkenazi Jews were first tested for the c.1906G>C founder mutation. Of the 240 families, 24, including Arabs and Jews from different ethnic origins, were tested positive. All tumors that lost expression of mismatch repair proteins also showed microsatellite instability. There was evidence for involvement of hMSH2 (15) hMLH1 (6) and hMSH6 (3) genes. Mutations were identified in 17/24 (71%) patients: 6 Ashkenazi families harbored the c.1906G>C mutation. Eleven other mutations (2 nonsense, 3 splice site and 6 small deletions) were detected. Three of the mutations are novel. No gross deletions or insertions were detected. This is the first report that characterizes the profile of HNPCC in a cohort of patients in Israel. Tumor testing indicated that the 3 main MMR genes are involved, and that mutation spectrum is broad. PMID- 18389389 TI - Differences in insulin sensitivity, pancreatic beta cell function and circulating adiponectin across glucose tolerance status in Thai obese and non-obese women. AB - Although adiponectin levels are associated with obesity and insulin insensitivity, the role of adiponectin in the progression to diabetes in non obese subjects is unclear. Therefore, 289 women aged 50-80 years without previous history of diabetes or impaired glucose tolerance (IGT) were studied. They were classified as normal glucose tolerance (NGT), IGT or diabetes based on WHO criteria. Insulin sensitivity (S) and beta cell function (B) indices were calculated using homeostasis model assessment (HOMA). In obese women with BMI > or = 25 kg/m(2) (n = 161), there were declines in HOMA-%S (P < 0.001), HOMA-%B (P < 0.05) and circulating adiponectin (P < 0.001) across glucose tolerance status. In non-obese women with BMI < 25 kg/m(2) (n = 128), there was no significant change in HOMA-%S in women with IGT and diabetes as compared to women with NGT. However, HOMA-%B (P < 0.05) and serum adiponectin levels (P < 0.001) were significantly decreased across glucose tolerance. Serum adiponectin levels were correlated to HOMA-%S in both obese and non-obese women while negative correlations between circulating adiponectin and HOMA-%B were demonstrated only in obese women. We have demonstrated in the present study the predominant role of beta cell dysfunction as compared to that of insulin resistance in the deterioration of glucose tolerance in non-obese women. Circulating adiponectin appears to be inversely related to beta cell dysfunction in addition to insulin resistance only in obese women. PMID- 18389390 TI - Influence of short- and long-term treadmill exercises on levels of ghrelin, obestatin and NPY in plasma and brain extraction of obese rats. AB - This study aims to clarify the effects of exercise on levels of appetite regulatory hormones in plasma and hypothalamus of obese rats. Diet-induced obese rats undergo short- (40 min) and long-term (40 min, 5 days/week for 8 weeks) exercises. The rats ran at a speed of 20 m/min on a 5 degrees slope treadmill. Rats undergoing short-term exercise were divided into C, E0, E1, E3, E12, and E24. Rats undergoing long-term exercise (LE) were compared to long-term control (LC). Concentrations of ghrelin, obestatin, and neuropeptide Y (NPY) were measured using radio immuno-assay. Expression of ghrelin receptor (GHSR-1a), putative obestatin receptor (GPR-39), and NPY in the hypothalamus was measured by quantitative RT-PCR. After short-term exercise, the plasma concentrations of ghrelin and obestatin were not changed, but NPY decreased. Ghrelin and obestatin in the hypothalamus decreased, and recovered 12 until 24 h. NPY increased and recovered after 24 h. Expression of GHSR-1a and NPY was not changed and GPR-39 was not observed. In LE, these changes are different in plasma and hypothalamus. It would be concluded appetite and body weight of obese rats are decreased by exercise through reduced level of ghrelin in the hypothalamus. Obestatin seems to have no effect in exercise-induced change in appetite. PMID- 18389391 TI - Alpha-1 antitrypsin: now available, but do we need it? AB - Severe alpha1-antitrypsin (AAT) deficiency is the best characterised genetic risk factor for the development of emphysema. AAT has a wide spectrum of antiprotease activity and its primary function is inhibition of neutrophil elastase in the lung. Smokers with this genetic defect develop severe impairment in their fifth to sixth decade of life. Intravenous administration of human AAT is well tolerated and has been shown to increase the levels of AAT in the alveolar lining fluid of individuals with this deficiency. In contrast to the proof of the biochemical effectiveness of augmentation treatment, the favourable clinical effect of AAT on pulmonary function, emphysema progression, morbidity and survival has not been persuasively demonstrated by prospective controlled clinical trials and remains controversial. PMID- 18389392 TI - Publication and non-publication of clinical trials: longitudinal study of applications submitted to a research ethics committee. AB - BACKGROUND: Not all clinical trials are published, which may distort the evidence that is available in the literature. We studied the publication rate of a cohort of clinical trials and identified factors associated with publication and nonpublication of results. METHODS: We analysed the protocols of randomized clinical trials of drug interventions submitted to the research ethics committee of University Hospital (Inselspital) Bern, Switzerland from 1988 to 1998. We identified full articles published up to 2006 by searching the Cochrane CENTRAL database (issue 02/2006) and by contacting investigators. We analyzed factors associated with the publication of trials using descriptive statistics and logistic regression models. RESULTS: 451 study protocols and 375 corresponding articles were analyzed. 233 protocols resulted in at least one publication, a publication rate of 52%. A total of 366 (81%) trials were commercially funded, 47 (10%) had non-commercial funding. 346 trials (77%) were multi-centre studies and 272 of these (79%) were international collaborations. In the adjusted logistic regression model non-commercial funding (Odds Ratio [OR] 2.42, 95% CI 1.14-5.17), multi-centre status (OR 2.09, 95% CI 1.03-4.24), international collaboration (OR 1.87, 95% CI 0.99-3.55) and a sample size above the median of 236 participants (OR 2.04, 95% CI 1.23-3.39) were associated with full publication. CONCLUSIONS: In this cohort of applications to an ethics committee in Switzerland, only about half of clinical drug trials were published. Large multi-centre trials with non commercial funding were more likely to be published than other trials, but most trials were funded by industry. PMID- 18389393 TI - Obesity in Switzerland: do estimates depend on how body mass index has been assessed? AB - BACKGROUND/AIM: In Switzerland monitoring of obesity in the general population is based on body mass index (BMI) derived from self-reported weight and height. This approach may lead to misclassification of obese subjects and misinterpretation of obesity prevalence and trends. In order to explore this potential bias, we compared studies with measured and self-reported data. METHODS: We analysed five studies based on measured BMI and five studies based on self-reported BMI, all of which were carried out in Switzerland between 1977 and 2004 and encompassed men and women aged 35-74 years. Obesity was defined as BMI>or=30 kg/m2. RESULTS: The prevalence of obesity was markedly higher (1.6 times) in studies with measured BMI in both sexes: 14.2% vs 8.8% in men and 12.5% vs 7.9% in women. These differences tended to increase with age in both sexes. However, a similar upward trend in the prevalence of obesity was observed with both methods (absolute increase per year in men and women respectively: 0.24% and 0.25% using measured BMI vs 0.17% and 0.20% using self-reported BMI). CONCLUSION: In Switzerland obesity prevalence in adults has clearly increased in the past three decades. Although the use of self-reported height and weight leads to a valid estimation of this increase, it results in a considerable underestimation of obesity prevalence rates in Switzerland. The type of assessment of height and weight should be taken into consideration when comparing prevalences of obesity between studies or regions or when using these prevalences to assess associated health risks or costs. PMID- 18389394 TI - Impact of an intervention to control risk associated with patient transfer. AB - QUESTION UNDER STUDY: Hospitals transferring patients retain responsibility until admission to the new health care facility. We define safe transfer conditions, based on appropriate risk assessment, and evaluate the impact of this strategy as implemented at our institution. METHODS: An algorithm defining transfer categories according to destination, equipment monitoring, and medication was developed and tested prospectively over 6 months. Conformity with algorithm criteria was assessed for every transfer and transfer category. After introduction of a transfer coordination centre with transfer nurses, the algorithm was implemented and the same survey was carried out over 1 year. RESULTS: Over the whole study period, the number of transfers increased by 40%, chiefly by ambulance from the emergency department to other hospitals and private clinics. Transfers to rehabilitation centres and nursing homes were reassigned to conventional vehicles. The percentage of patients requiring equipment during transfer, such as an intravenous line, decreased from 34% to 15%, while oxygen or i.v. drug requirement remained stable. The percentage of transfers considered below theoretical safety decreased from 6% to 4%, while 20% of transfers were considered safer than necessary. A substantial number of planned transfers could be "downgraded" by mutual agreement to a lower degree of supervision, and the system was stable on a short-term basis. CONCLUSION: A coordinated transfer system based on an algorithm determining transfer categories, developed on the basis of simple but valid medical and nursing criteria, reduced unnecessary ambulance transfers and treatment during transfer, and increased adequate supervision. PMID- 18389395 TI - New perspectives on subjective memory complaints. PMID- 18389396 TI - A study into the psychosocial determinants of perceived forgetfulness: implications for future interventions. AB - Many healthy individuals perceive themselves as forgetful and are interested in interventions to decrease their worries and increase their memory functioning. Educational interventions can be more effective when determinants are targeted that are known to predict perceived forgetfulness. In the present study, first, a broad range of determinants was selected from the literature and from experiences in clinical settings and, second, the most important determinants among the selected ones were identified with multivariate regression analyses. The study had a cross-sectional design. A sample of 300 healthy participants aged over 54 years filled in a self-report questionnaire. Findings indicated that low memory self-efficacy, high memory-related anxiety, negative attitude and high subjective norm (e.g. anticipating negative evaluations by important others) were the most important correlates of perceived forgetfulness. It is argued that future interventions should focus on the specified determinants to improve programme effectiveness in reducing subjective memory complaints. PMID- 18389397 TI - Memory complaints in healthy young and elderly adults: reliability of memory reporting. AB - The relationship between memory complaints and objective memory performance remains poorly understood, particularly in young and middle aged people. We studied the relationship between reports of memory complaints and objective memory performance, and the possibility of differentiating good and poor reporters across the lifespan based on concordance between reported abilities and objectively assessed performance. This cross-sectional study enrolled 292 healthy individuals, aged 18 to 87 years, able to perform common activities of daily living and without neurological or psychiatric conditions or systemic diseases likely to interfere with cognition. No correlation between memory complaints, as assessed by the Subjective Memory Complaints scale (SMC) score and the objective memory performance, evaluated by the long-delay free recall (LDFR) score of the California Verbal Learning Test (CVLT), was found, even when grouping the participants by decade. The SMC score was influenced by the presence of depressive symptoms. Participants who were more educated, female and younger tended to have a higher CVLT-LDFR score. Younger subjects were more likely to have good memory performance and report few memory complaints than older subjects. In conclusion, there are differences in the reliability of memory reporting across the lifespan, younger subjects being more likely to correctly report good memory than older subjects. PMID- 18389398 TI - The influence of lifetime depression on self-reported memory and cognitive problems: results from the National Comorbidity Survey-Replication. AB - The current study investigated the association between lifetime major depressive disorder (MDD) and self-reported memory and cognitive problems in a general population sample. The study was based on data from the National Comorbidity Survey-Replication (n = 5692). The relationship between lifetime MDD and self reported memory and cognitive problems was examined while controlling for other 1 year and lifetime psychiatric disorders. We found a lifetime history of depression, but no other lifetime psychiatric disorder, to be associated with self-reported memory and cognitive problems. We review the results in relation to theories regarding the comorbidity of depression and cognitive problems. A history of depression increases the likelihood that individuals will make negative evaluations of their memory and cognitive functioning. Though depression is highly comorbid with other disorders, our results are unique in demonstrating the specificity of depression in its association with self-reported memory and cognitive problems. The observed association between depression and self-reported memory and cognitive problems may reflect that depression increases the risk for cognitive decline or may reflect that individuals with a history of depression tend to rate their memory as poor, or both. PMID- 18389399 TI - Social networks and depressive symptoms among elderly women and men in Havana, Cuba. AB - OBJECTIVES: To examine the main and the stress-buffering effects of social networks on depressive symptoms among elderly Cuban men and women living in La Havana. METHOD: Information was gathered from a representative sample of the elderly population in Havana (n = 1905), as part of the SABE (Salud, Bienestary Enuejecimiento) study. Depressive symptoms were measured using the 15-item Geriatric Depression Scale. The structure and function of social networks were studied. Gender-specific multivariate logistic regressions were fitted to test the main (independent of stressors) and the stress-buffering effects (in the presence of financial strain or disabilities) on depressive symptoms. RESULTS: Social ties were associated with a lower prevalence of depressive symptoms in women and men independently of the presence of stressors. Women who were or had been married, lived in an extended family, and enjoyed balanced exchanges with relatives and children reported low prevalence of depressive symptoms. Men were less likely to report depressive symptoms if they were currently married, and did not live alone. Social networks buffered the effect of financial strain on depression, but not in the event of disability. CONCLUSION: In Cuba, networks centered on children and extended family were associated with low frequency of depressive symptoms, ruling contrary to common findings in developed societies. These living arrangements have an important role in buffering the impact of financial strain on depressive symptoms. PMID- 18389400 TI - Assessing the validity and reliability of the Pool Activity Level (PAL) Checklist for use with older people with dementia. AB - Activity is key to maintaining physical and mental health and well-being. However, as dementia affects the ability to engage in activity, care-givers can find it difficult to provide appropriate activities. The Pool Activity Level (PAL) Checklist guides the selection of appropriate, personally meaningful activities. The aim of this study was to assess the reliability and validity of the PAL Checklist when used with older people with dementia. A postal questionnaire sent to activity providers assessed content validity. Validity and reliability were measured in a sample of 60 older people with dementia. The questionnaire response rate was 83% (102/122). Most respondents felt no important items were missing. Seven of the nine activities were ranked as 'very important' or 'essential' by at least 77% of the sample, indicating very good content validity. Correlation with measures of cognition, severity of dementia and activity performance demonstrated strong concurrent validity. Inter-item correlation indicated strong construct validity. Cronbach's alpha coefficient measured internal consistency as excellent (0.95). All items achieved acceptable test-retest reliability, and the majority demonstrated acceptable inter-rater reliability. We conclude that the PAL Checklist demonstrates adequate validity and reliability when used with older people with dementia and appears a useful tool for a variety of care settings. PMID- 18389401 TI - Movement in mind: the relationship of exercise with cognitive status for older adults in the Swedish National Study on Aging and Care (SNAC). AB - The purpose of this study was to investigate the relationship of light and strenuous exercise, and self-reported change in exercise status, with different components of cognitive function, and gender differences in this relation, in a large, representative sample included in the Swedish National study on Aging and Care (SNAC). Eight-hundred-and-thirteen participants in age-cohorts from 60-96 years completed a wide range of cognitive function tests, the Mini Mental State Exam (MMSE) and survey questions concerning exercise behaviour and exercise change with light or strenuous intensity. ANCOVA, controlling for age, education, depression, functional status and co-morbidity, demonstrated a main effect for light exercise, but not for strenuous exercise, on five of the six cognitive tests and the MMSE, for men but nor for women. A negative change in exercise status was associated with lower MMSE scores for men but not for women. Individuals exercising with light intensity several times a week had the highest cognitive test and MMSE scores and the inactive group had the lowest scores. The results of the study may contribute to increased knowledge in the exercise-mental health relationship for elderly and spawn new research specifically on gender differences in this relation. PMID- 18389402 TI - Genetic and environmental influences on depressive symptoms by age and gender in African American twins. AB - Depression is typically considered relative to individuals and thought to originate from both biological and environmental factors. However, the environmental constraints and insults that African Americans experience likely influence the concordance by age and gender for depression scores among adult African American twins. Monozygotic (MZ) (n = 102) and Dizygotic (DZ) (n = 110) twins, age 25-88 years in the Carolina African American Twin Study of Aging were examined using an 11-item version of the CES-D measure of depressive symptomatology. Those participants with scores above nine were considered depressed. Overall, the MZ pairs had a higher concordance than the DZ pairs implying genetic influence. Both MZ and DZ males had higher concordances than either female zygosity groups. The difference between the concordance rates for MZ and DZ twin pairs was greater in males than females. By age group, the difference between the concordance rates for younger MZ and DZ twin pairs was much larger than for older pairs. The results suggest that, even though African Americans may be at risk for depression due to contextual/environmental factors, genetic influences remain important. PMID- 18389403 TI - Knowledge and skills of professional carers working with older people with depression. AB - OBJECTIVES: The current study was designed to evaluate the knowledge, skills and self-efficacy of care providers from the perspective of professionals working in the aged-care industry. METHOD: Participants were 21 professional carers, 10 General Practitioners and 7 aged-care managers. Focus groups, which involved the completion of a semi-structured interview related to knowledge, recognition, confidence, referral procedures and use of screening tools for the detection of depression, were conducted. RESULTS: The results showed that all groups of respondents recognised significant gap in the knowledge and awareness of depression among professional care staff working with older people in both the community and residential care-settings. Skills in the detection and monitoring of depression and the self-efficacy of these care staff were also seen to be a problem. DISCUSSION: The implications of these findings in terms of training programmes for professional carers working in the aged health care sector are discussed. PMID- 18389404 TI - Strain in nursing care of people with dementia: nurses' experience in Australia, Sweden and United Kingdom. AB - OBJECTIVES: The aim of this study was to explore nurses' experience of strain in dementia care. METHOD: Focus groups were held with 35 nurses in Sweden, Australia and UK, who care for people with dementia. The discussions were tape-recorded and analysed using qualitative content analysis. RESULTS: The nurses described the complexity of their situation and referred to environmental factors such as 'the system', community attitudes, other staff, residents' family members and also their own family. With regard to caring for people with dementia, three main sources of strain could be identified: Being unable to reach; Trying to protect; and Having to balance competing needs. CONCLUSION: The nurses' experience could be understood as a desire to do the best for the people in their care by trying to alleviate their suffering and enhance their quality of life. When they did not have the resources, opportunity or ability to do this, it caused strain. PMID- 18389405 TI - Screening for depression and assessing change in severity of depression. Is the Geriatric Depression Scale (30-, 15- and 8-item versions) useful for both purposes in nursing home patients? AB - The objectives of this study were to determine the ability of the 30-, 15- and 8 item versions of the GDS for screening and assessing change in severity of depression in nursing home patients. The GDS and the MADRS were administered to 350 elderly NH-patients by trained interviewers. The presence of major (MaD) or minor depression (MinD) was evaluated with the Schedules for Clinical Assessment in Neuropsychiatry. Receiver Operator Characteristic (ROC) curves of the GDS versions were performed to measure the ability to screen on depression. The ability to measure change in severity of depression was measured by differences in mean GDS-scores and mean MADRS-scores between patients with MaD, MinD and no depression, and expressed in terms of effect sizes. It was found that in ROC curves all three GDS-versions performed well. The MADRS showed larger effect sizes for the differences between MaD, MinD and no depression than the GDS versions. The effect sizes of the three GDS versions were comparable. We conclude that all three versions of the GDS can be used for screening on depression among NH-patients. The MADRS is superior to the GDS for assessment of (changes in) severity of depression, but the GDS also appears to be an acceptable instrument for this purpose and is less time-consuming. PMID- 18389406 TI - Response variability of salivary cortisol among older adults under psychological stress. AB - In this study, a quasi-experimental design with repeated measures was used to compare anxious (n = 129) and nonanxious (n = 186) older adults on the cortisol secretion rate attributable to an experimental stressor. Our results support the hypothesis that a first-order longitudinal factor model appropriately describes the cortisol concentration in three saliva samples collected at two experimental times. The model tested explained between 82.6 and 98.0% of the variance in cortisol concentration of the respondent's saliva samples at T1, and between 55.4% and 78.4% at T2. In the nonanxious group ((State-Trait Anxiety Inventory) STAI < 42), the magnitude of the experiment-related cortisol reactivity was, respectively 51 and 33% higher than the respondent's baseline reactivity level at both T1 and T2. By contrast, in the anxious group (STAI >or= 42), our results showed no significant gradient in the magnitude of the cortisol reactivity at either time. This result was interpreted to be in agreement with the helplessness reaction hypothesis. These results suggest that salivary cortisol is a valid measure that is sensitive to experimental stress, and may therefore, be useful in examining physiological response to stress. PMID- 18389407 TI - Knowledge of memory aging and Alzheimer's disease in college students and mental health professionals. AB - In this study, college students and mental health professionals completed the Knowledge of Memory Aging Questionnaire, Alzheimer's Disease Knowledge Test and the Fraboni Scale of Ageism before and after a lecture on normal and pathological memory issues in adulthood. Results confirmed that professionals were more knowledgeable about memory aging and Alzheimer's disease (AD) and less ageist than college students. Analyses of pre- and post-lecture response accuracy yielded comparable benefits in memory aging and AD knowledge for both groups. Correlation analyses provided modest evidence for the influence of ageist attitudes on the knowledge measures. Implications for memory education programs and psychology curriculum are considered. PMID- 18389408 TI - The impact of spirituality on health-related quality of life among Chinese older adults with vision impairment. AB - The goals of this study were to examine the impact of spirituality on both generic and vision-specific health-related quality of life (HRQOL) and the role of spirituality in the linkage between vision impairment and HRQOL outcomes among Chinese older adults with visual problems. A sample of 167 urban older adults with visual problems were interviewed using a structured questionnaire to assess self-reported visual functioning, general health, spirituality and both generic and vision-specific HRQOL. Objective visual function was clinically measured by ophthalmologists in terms of distance visual acuity. It was found in the study that spirituality was positively associated with both general physical health and general mental health, with a higher correlation with general mental health than that with general physical health. Moreover, spirituality was associated significantly with vision-specific HRQOL, including multiple domains. The results showed that spirituality played a mediating role in the linkage between vision impairment and general mental health. These results suggest that spirituality is significantly associated with multiple domains of HRQOL and may play an important role in the process of adaptation to age-related vision loss among Chinese older adults with vision impairment. PMID- 18389409 TI - Activities enjoyed by patients with dementia together with their spouses and psychological morbidity in carers. AB - Caring for a spouse with dementia is stressful and respite care is sometimes used to reduce this burden. Spouses may find some aspects of caring rewarding but the literature on positive aspects of caring is limited. To describe activities enjoyed by patients with dementia together with their spouses, and examine their relationship with psychological morbidity in carers. A convenience sample of 46 patients with mild to moderate dementia (91% with Alzheimer's disease, AD) and their spouses were interviewed at home. Spouses completed the Pleasant Events Schedule (PES-AD) to identify activities enjoyed by patients and spouses on their own and together. Psychological morbidity in spouses was assessed using the General Health Questionnaire (GHQ-12). Cognitive functions, and non-cognitive symptoms were also assessed in patients. Multiple regression analysis using age, Mini-Mental State Examination, Cornell Scale for Depression in Dementia, Revised Memory and Behaviour Problems (RMBP) checklist frequency, and PES-AD- together scores as independent variables found PES-AD-together and RMBP-frequency to be independent predictors of GHQ-12 scores in spouses, but the model could explain only 28% of variance. Facilitating activities that are enjoyed by both patients with dementia and spouses may be an alternative intervention strategy to reduce carer burden. PMID- 18389410 TI - The relationship between pre-fitting expectations and willingness to use hearing aids. AB - Hearing-aid use in the elderly population is problematic since not all people who would benefit from hearing aids actually obtain one. In this study, the relationship between pre-fitting expectations and willingness to use hearing aids is addressed. One hundred adult hearing aid candidates (mean age 68.6 years) were randomly recruited from eight different private ENT practices. The subjects completed a questionnaire comprising a number of different aspects of expectation and some additional variables potentially influencing willingness to use hearing aids. Based on the outcome of the questionnaire a linear regression model predicting motivation to use hearing aids was calculated and evaluated. Among the 11 parameters evaluated three contributed significantly to the model of willingness. The three predictor variables were expectations towards improvement of quality of life, stigmatization, and self-rated hearing ability. They accounted for about 55% of the variability in the data for willingness. Examination of a sub-sample three months after the survey had taken place revealed a distinct relationship between willingness and the decision to obtain or decline hearing aids. In conclusion, the results strongly encourage supporting positive expectations in order to motivate individuals with hearing impairment to use hearing systems. PMID- 18389411 TI - Questionnaire investigation of musicians' use of hearing protectors, self reported hearing disorders, and their experience of their working environment. AB - Musicians in symphony orchestras are exposed to harmful sound levels. Although research shows that industrial workers have a higher propensity to noise-induced hearing loss, musicians can also develop a hearing loss from noise exposure. Furthermore, musicians can suffer from tinnitus, hyperacusis, and distortion, among other hearing disorders, which can affect their work more severely than a hearing loss. This study investigated the use of hearing protectors, the prevalence of self-reported hearing disorders among musicians, and the importance of these hearing disorders to the musicians. The musicians at three Danish symphony orchestras were asked to complete a questionnaire on the topic. Results showed that Danish musicians are aware of the dangers of loud music, yet they rarely use hearing protectors and not always correctly; however, musicians with hearing disorders use hearing protectors more frequently. In addition, the musicians questioned suffered from different hearing disorders. Education is needed to change musicians' opinion of hearing conservation and hearing protectors. The education must be directed to both the musicians and the administration of the symphony orchestras. PMID- 18389412 TI - Measuring outcome from Vestibular Rehabilitation, Part I: Qualitative development of a new self-report measure. AB - Research suggests that Vestibular Rehabilitation (VR) is an effective treatment for dizziness, but there is currently no measure specifically designed to assess treatment outcome. A review of existing self-report measures of dizziness indicates that no measure has been designed for longitudinal application and all suffer from limitations which restrict their usefulness in measuring VR outcome. A need for a psychometrically robust patient-oriented measure of quality of life benefit from VR is identified. The aim of the present study was to explore dimensions relevant to VR with a view to developing a measure of outcome. Eighteen adults receiving VR participated in interviews about the quality of life impact of dizziness. Qualitative analysis revealed 64 themes describing self perceived quality of life impact. Themes were developed into potential questionnaire items and 35 were selected to represent the quality of life impact of dizziness in a prototype questionnaire. A quarter of items in the prototype questionnaire refer to issues not addressed by existing measures; the remaining items draw together issues covered by the range of questionnaires currently in use. PMID- 18389413 TI - Hearing preservation using combined monitoring of extra-tympanic electrocochleography and auditory brainstem responses during acoustic neuroma surgery. AB - The aim of this study was to evaluate the efficacy of an intraoperative monitoring hearing preservation strategy that includes simultaneous recordings of an auditory brainstem response (ABR) and non-invasive electrocochleography (ECochG). The combined ABR and tympanic membrane (TM) ECochG testing was performed in 74 patients undergoing acoustic neuroma (AN) surgery. In addition, EMG recordings were conducted to monitor the facial nerve function. Hearing was preserved in 19 of the 30 patients with residual hearing prior to surgery (63%), and facial nerve function was maintained in 89% of the patients. In most cases, the presence of both auditory brainstem and TM-ECochG responses at the end of surgery was associated with preservation of postoperative functional hearing; however, eight patients had a TM-ECochG response with a complete loss of the ABR, pointing to deafferentation of the auditory nerve. Tumour size and preoperative hearing thresholds significantly affected the postoperative hearing. The TM ECochG response yielded large reproducible responses, which, in some patients, was the only way to monitor the auditory function. This auditory monitoring approach offers a valuable auditory tool that helps to improve the hearing preservation during AN surgery. PMID- 18389414 TI - Learning effect observed for the speech reception threshold in interrupted noise with normal hearing listeners. AB - Traditionally, the speech reception threshold (SRT) for sentences in noise is measured in stationary speech-shaped noise; however, non-stationary masking noises are gradually becoming more common. A previous study by Rhebergen et al. (2006) suggested that a learning effect might be present for the SRT in interrupted noise but not in stationary noise. The current study tested if a stable SRT for female or male speech is reached by determining SRTs in listeners with normal hearing in either stationary or 8-Hz interrupted noise after five replications. Contrary to repeated SRT measurements in stationary noise, a significant improvement was observed for SRTs in interrupted noise with replication. For both speech materials, after five replications, the SRT in interrupted noise improved about 3 to 4 dB in comparison to the first SRT. Stable thresholds seem to be reached after about two replications. This experiment shows that there is a substantial learning effect present in SRT measurements with interrupted noise but not in stationary noise. With non-stationary noise, it is recommended to include a repeated measures design. PMID- 18389415 TI - Discrimination of release time constants in hearing-aid compressors. AB - We examined the ability of twenty-five hearing-impaired and eight normal-hearing listeners to discriminate between release time constants used for compression in hearing aids. The compressor was a standard three-channel system. The stimuli were normal and 'vocoded' sentences from a male and female database. In agreement with other studies looking at different outcomes, performance varied greatly across individuals. This variation was greater in hearing-impaired listeners, for whom the discriminability of a release time of 5 ms from one of 5000 ms (with the attack time fixed at 5 ms) ranged from chance to perfect. This variability was not significantly related to hearing impairment nor to individuals' compression ratios. PMID- 18389416 TI - Cochlear implantation in deaf children with associated disabilities: challenges and outcomes. AB - The issue of cochlear implantation in deaf children with associated disabilities is an emerging subject. Currently, there is no consensus on whether to implant children with multiple impairments; moreover, it may be difficult to evaluate these children with standard tests pre- or post-implantation. In addition, these children often have poor speech perception and language skills, making assessment more difficult. Despite these factors, these children often receive important benefits in daily life, with an overall improvement in quality of life. In the present study, post-implant outcomes of 23 profoundly deaf children with neuropsychiatric disorders were analysed, using objective measures of speech perception, and a questionnaire administered to the parents, aimed at evaluating the benefits in daily life after implantation. The results were quite variable, but overall positive, in terms of speech perception, communication abilities, and improvement in quality of life. The findings add an additional piece of evidence to support the effectiveness of cochlear implantation in these special cases. PMID- 18389417 TI - Tympanometry in rhesus monkeys: effects of aging and caloric restriction. AB - Caloric restriction is the only known method of increasing lifespan in laboratory animals. The present study was conducted as part of a larger investigation into the effect of caloric restriction on longevity of rhesus monkeys as a model for human aging. This study focused on the effects of caloric restriction and aging on measures of middle-ear function measured with tympanometry. Peak compensated static acoustic admittance (peak Y(tm)) tended to be reduced with aging. For tympanometric width (TW), the effect of age was significant with TW increasing with age. Males had a trend of narrower TW than females. A significant age by sex interaction indicated that TW for males stays relatively constant, whereas TW for females increases with age. The equivalent ear canal volume (V(ea)) was significantly larger in male monkeys than in female monkeys, and marginally larger for the control monkeys than for the caloric restricted monkeys. These results parallel many findings in middle-ear function in aging humans. Longitudinal studies are planned. PMID- 18389418 TI - Topical review: temporomandibular disorders in an integral otic symptom model. AB - The literature has closely observed otic symptoms (and other craniofacial complaints) in temporomandibular disorders; however, there is little evidence for an association between the two. This review tries to provide an integrated biological basis for otic symptoms in temporomandibular disorders from both anatomical and physiological points of view; it also attempts to enlarge the view of one of the ranges of central and peripheral mechanisms involved. The pathophysiology of common symptoms is integrated within different health specialties through basic science. This review is not based on a structured selection of randomized controlled trials; rather, it deals with perspectives of otic symptoms triggered or exacerbated by stomatognathic dynamics. PMID- 18389420 TI - Corticotropin-releasing factor (CRF) in stress and disease: a review of literature and treatment perspectives with special emphasis on psychiatric disorders. AB - The CRF family of neuropeptides and receptors is involved in a variety of stress responses, in the regulation of appetite, metabolic and inflammatory processes as well as intestinal movements. From a primarily psychiatric perspective, the present paper reviews the literature on its anatomy, physiology and its involvement in psychiatric, neurological and inflammatory diseases. Finally, recent developments in the pharmacological aspects of CRF in these diseases are reviewed. PMID- 18389421 TI - Long-term outcome of patients with dysthymia and panic disorder: a naturalistic 9 year follow-up study. AB - The highly prevalent psychiatric disorders dysthymia and panic disorder have often a chronic or recurrent course with superimposed major depression. The prominent comorbidity between these diagnoses constitutes a confounding factor in the study of long-term outcome. We performed a 9-year follow-up of 38 patients with "pure" diagnoses, i.e. without comorbid dysthymia and panic disorder, selected from two 2-year naturalistic treatment studies with psychotherapy and antidepressant medication. The aims of the present study were to investigate 1) the stability of change, and 2) the impact of comorbid personality disorders (PDs) on long-term outcome. Patients were reassessed with SCID-I and SCID-II interviews, SCL-90/BSI and a detailed, modified life-charting interview, investigating course and treatment over time. About 50% of patients showed substantial improvement, of whom about half were in remission. Comorbid PD was a negative prognostic factor independently of Axis I diagnosis. Although patients with panic disorder had a lower frequency of comorbid PD, later onset, shorter duration of illness and better outcome after the original studies, there was no difference in the long-term outcome. The less stable outcome among panic patients suggests that standard treatments are not resulting in enduring remission. In order to achieve remission, it is necessary to 1) address comorbid PDs, 2) perform careful assessments of all comorbid diagnoses, and 3) build routines for the follow-up and augmentation of treatments. PMID- 18389422 TI - Disentangling dysthymia from major depressive disorder in suicide attempters' suicidality, comorbidity and symptomatology. AB - Dysthymia and major depressive disorder (MDD) are both risk diagnoses for suicidal behaviour. The aim of the present study was to identify clinical differences between these disorders, with a special reference to dysthymia. We studied suicidal behaviour, comorbidity and psychiatric symptoms of inpatient suicide attempters with dysthymia and MDD. We used DSM III-R diagnostics, the Suicide Assessment Scale (SUAS) and the Comprehensive Psychopathological Rating Scale (CPRS), part of which is the Montgomery and Asberg Depression Rating Scale (MADRS). Suicide mortality, number of repeated suicide attempts, method of suicide attempt and comorbidity of Axis I did not differ between the groups. Dysthymia patients, however, suffered more than MDD patients from DSM-III-R Axis II diagnoses (above all cluster B). There was no significant difference in Axis III comorbidity. Total SUAS, CPRS and MADRS scores did not differ significantly between the groups. When studying separate SUAS and CPRS items in a multivariate analysis, the CPRS items "aches and pains", "increased speech flow", increased "agitation" and "less tendency to worrying over trifles" as well as young age remained independently associated with dysthymia. Dysthymia patients, who later committed suicide, more often reported increased "aches and pains" than those who did not commit suicide. In this small sample of suicide attempters, we conclude that dysthymia suicide attempters, more often than MDD patients, have a comorbidity with personality disorders, which combined with a picture of aches and pains, could be factors explaining their suicidality. PMID- 18389423 TI - Comparing psychogeriatric units to ordinary long-term care units - are there differences in case-mix or clinical symptoms? AB - This study is a comparative analysis of long-term psychogeriatric and mixed-care unit patient characteristics in nursing homes and hospitals in Helsinki. The role of the so-called psychogeriatric nursing homes is still under question and not well developed. The aims of the study were to identify and survey psychogeriatric wards and analyze whether these units differ from the rest of the long-term units/wards in terms of patients' characteristics. Participants studied were a total of 2828 elderly long-term care residents, of which 372 were living in long term psychogeriatric and 2456 in ordinary long-term care units. Data were drawn from the Resident Assessment Instrument (RAI) database and had been collected in the project "Benchmarking and implementation of RAI in Elderly Care in Finland". Scales and items based on Minimum Data Set were used for the comparisons. Resident groups in psychogeriatric units and in ordinary, mixed-client settings were clearly distinguishable. The psychogeriatric residents were younger, had more comorbidity as to psychiatric diseases, and had more often psychiatric symptoms and psychotropic medications. The residents in psychogeriatric units did not differ in cognitive and functional status from those in mixed-client units and had similar comorbidity as to somatic diseases. This study shows that psychiatric symptoms that need to be addressed are common in long-care facilities. The results emphasize the importance of recognition of the multiple care needs of elderly with severe mental symptoms or illness. PMID- 18389424 TI - Treatment of obsessive-compulsive disorder: personal follow-up of a 10-year material from an outpatient county clinic. AB - Individuals who suffer from obsessive-compulsive disorder (OCD) experience obsessive thoughts and/or urges to engage in compulsive behaviours. The condition causes severe discomfort and, in many cases, leads to serious impairment in social and work-related functioning. During the past decade, the Department of Psychiatry at Forde Central Hospital has systematically offered a combination of exposure with response prevention and antidepressant medication to these patients. In this paper, we describe our treatment model, including two case histories, and evaluate the outcome of the first 25 patients, who were included in this prospective study. They were evaluated at the onset, at the end of treatment and at follow-up, 1-10 years after treatment cessation. We employed the Yale-Brown Obsessive Compulsive Scale (Y-BOCS), Obsessive-Compulsive Checklist (OCC) and Beck Depression Inventory (BDI). The mean scores for obsessions, compulsions and depression were significantly reduced after treatment. Further improvement occurred during the follow-up period. Effect sizes were above 1.3 on all instruments. Eleven participants were gainfully employed prior to treatment, 14 at the end of treatment and 18 at follow-up. The study included few participants, and rates of improvement were not compared with a control group. We therefore cannot conclude that the positive changes were caused by the treatment, but lasting spontaneous remission among patients suffering from OCD is rare. The study shows that a county outpatient psychiatric unit can utilize methods that have previously been shown to be effective in highly specialized treatment facilities, with good results. PMID- 18389425 TI - Do changes on MCMI-II personality disorder scales in short-term psychotherapy reflect trait or state changes? AB - The Millon Clinical Multiaxial Inventory (MCMI) has become an important and commonly used instrument to assess personality functioning. Several studies report significant changes on MCMI personality disorder scales after psychological treatment. The aim of the study was to investigate whether pre-post treatment changes in 39-session psychodynamic group psychotherapy as measured with the MCMI reflect real personality change or primarily reflect symptomatic state changes. Pre-post-treatment design included 236 psychotherapy outpatients. Personality changes were measured on the MCMI-II and symptomatic state changes on the Symptom Check List 90-R (SCL-90-R). The MCMI Schizoid, Avoidant, Self defeating, and severe personality disorder scales revealed substantial changes, which could be predicted from changes on SCL-90-R global symptomatology (GSI) and on the SCL-90-R Depression scale. The MCMI Dependent personality score was the only MCMI personality scale showing significant change when the SCL-90-R Depression change score was included as a covariate. Splitting patients into those with and without personality disorders did not change the results. Observed changes on MCMI-II personality disorder scales in short-term psychotherapy reflect change in symptomatic state. The MCMI-II Base Rate cut-off points probably include too many patients, justifying the introduction of new scoring procedures in the MCMI-III. PMID- 18389426 TI - Climatic factors and bipolar affective disorder. AB - In bipolar disorder, the factors provoking a new episode are unknown. As a seasonal variation has been noticed, it has been suggested that weather conditions may play a role. The aim of the study was to elucidate whether meteorological parameters influence the development of new bipolar phases. A group of patients with at least three previous hospitalizations for bipolar disorder was examined every 3 months for up to 3 years. At each examination an evaluation of the affective phase was made according to the Hamilton Depression Scale (HAM-D(17)), and the Bech-Rafaelsen Mania Rating Scale (MAS). In the same period, daily recordings from the Danish Meteorological Institute were received. We found no correlations between onset of bipolar episodes [defined as MAS score of 11 or more (mania) and as HAM-D(17) score of 12 or more (depression)] and any meteorological parameters. We found a statistical significant correlation between mean HAM-D(17) scores and change in mean and maximum temperature, and non statistical significant correlations between mean MAS scores and rainfall plus atmospheric pressure, and non-statistical significant correlations between mean HAM-D(17) scores and hours of sunshine and cloudiness. Though meteorological factors may have an impact on triggering new episodes in bipolar patients, they do not constitute a dominant cause. PMID- 18389427 TI - Behaviour, attention and cognition in severe dyslexia. AB - Information about coexisting problems in dyslexia may be vital for diagnostic and remedial procedures. There are few studies in this area, and to our knowledge none for the students in focus in this study. They all have phonological deficits and severely impaired reading abilities, in spite of prolonged educational remediation. The aim was to assess if these students have more behavioural/emotional problems than normal reading students. A clinical sample of 70 students, 59 males and 11 females, were compared to a normal reading control group. The groups were pair-wise matched on age, gender, cognitive level, and whether they lived in rural or urban areas. Mean age for the two groups was 150 months, and mean IQ scores approximately 100. Information on behaviour/emotions was obtained from parents, teachers and participants by means of the Child Behavior Checklist, Teacher's Report Form and Youth Self Report. The dyslexia group showed significantly more problems in all areas than the controls. This was reported from all three groups of informants. Parents reported most problems, internalizing behaviour for more than 50% of the participants and total problem behaviour for nearly 45%. Significantly more attention problems were also reported for the dyslexia group from all informants. Parents reported that nearly half the group demonstrated attention problems. Both groups of participants had more problems reported for those with IQ scores of 100 or lower. Further research is needed in this area, but the results clearly indicate that identifying additional behavioural/emotional problems may be imperative for students with severe reading problems. PMID- 18389428 TI - The Adolescent Adjustment Profile (AAP) in comparisons of patients with obesity, phenylketonuria or neurobehavioural disorders. AB - Psychosocial development in children with chronic disease is a key issue in paediatrics. This study investigated whether psychosocial adjustment could be reliably assessed with the 42-item Adolescent Adjustment Profile (AAP) instrument. The study mainly focused on adjustment-to-obesity measurement, although it compared three patient groups with chronic conditions. All phenylketonuria (PKU) patients in Sweden between ages 9 and 18 and their parents and teachers were invited to participate. Patients with neurobehavioural syndromes and obesity were age- and gender-matched with PKU patients. Healthy children constituted a reference group. Psychosocial adjustment was measured using the AAP, which is a multi-informant questionnaire that contains four domains. Information concerning parents' socio-economic and civil status was requested separately. Respondents to the three questionnaires judged the PKU patients to be normal in all four domains. Patients with neurobehavioural syndromes demonstrated less competence and the most problems compared with the other three groups. According to the self-rating, the parent rating and the teacher rating questionnaires, obese patients had internalizing problems. The parent rating and the teacher rating questionnaire scored obese patients as having a lower work capacity than the reference group. Compared with the reference group, not only families with obese children but also families with children with neurobehavioural syndromes had significantly higher divorce rates. Obese patients were also investigated with the Strength and Difficulties Questionnaire (SDQ), another instrument that enables comparison between two measures of adjustment. The AAP had good psychometric properties; it was judged a useful instrument in research on adolescents with chronic diseases. PMID- 18389429 TI - Coercion in patients who at their first contact with the psychiatric services system were diagnosed within the schizophrenia-spectrum in Denmark. A register study. AB - The level of use of coercive measures in patients diagnosed with a schizophrenia spectrum disorder at their first contact with the psychiatric services system in Denmark is not known. The aim of the study was to investigate the level of use of coercive measures during first year of contact in this group of patients. Using the longitudinal national registers, the use of coercive measures for each individual was calculated in a 1-year period from 1 January 1999 to 31 December 2001, for patients in Denmark who at their first contact with the psychiatric services system were diagnosed within the schizophrenia-spectrum (F2 in ICD 10); 2222 patients were identified. The mean age was 30.7 years and 63.6% were males; 554 patients (24.9%) experienced use of coercive measures. A total of 222 patients (10.0%) were admitted involuntarily, 308 (13.9%) were detained. Seventy two patients (3.2%) were treated involuntarily with anti-psychotic medication and 158 (7.1%) with sedative medication. A total of 241 patients (10.8%) were restrained with a leather belt, and 84 patients (3.8%) with a leather belt and strap(s). The mean duration of involuntary admission was 30.3 days. The mean duration of restraint with a leather belt was 2.66 days. This study provides information about the level of use of coercive measures in first-episode schizophrenia-spectrum disorders in Denmark. PMID- 18389430 TI - Medical pluralism. PMID- 18389431 TI - The experience of low back pain in Iranian women: a focus group study. AB - Chronic low back pain (CLBP) often proves resistant to treatment because there are different biological, psychological, and social factors that can affect its treatment. In this study, we explored Iranian women's beliefs regarding the cause of their LBP. Findings are based on four focus groups (N = 24 participants) with LBP patients. Participants articulated their causation beliefs (Explanatory Models) about low back pain through three themes: idioms of distress: stress in women's lives; lack of knowledge resulted in high-risk behaviors; and organic/biomedical. Given the difficultly regarding diagnosis and treatment of CLBP, understanding patients' health beliefs, causation beliefs, and experiences are warranted. PMID- 18389432 TI - "Can it be that god does not remember me": a qualitative study on the psychological distress, suffering, and coping of Dominican women with chronic filarial lymphedema and elephantiasis of the leg. AB - The psychological states of Dominican women with chronic lymphedema and elephantiasis of the leg and the coping strategies they used to ameliorate the negative psychological effects of this condition were explored using modified precepts of grounded theory method. Qualitative data were gathered through in depth interviewing and focus group discussions held in the Dominican Republic. Thematic results found that compounding their physical disfigurement, functional limitations, and social losses were feelings of depression, embarrassment, social isolation, and despair. Adaptive problem solving and emotion-focused coping strategies that emerged during analysis also are discussed. It is recommended that management of psychological distress should be a significant component of lymphedema management programs in developing countries. PMID- 18389433 TI - A qualitative study examining psychosocial distress, coping, and social support across the stages and phases of epithelial ovarian cancer. AB - Ovarian cancer patients experience high levels of anxiety and depression, yet there is little research regarding coping and support of this population. In this study we examined the experiences of women during diagnosis and treatment via 30 semistructured interviews. The interviews were analyzed qualitatively, and five main themes were evident: (1) extreme blunting; (2) having a "forgotten cancer"; (3) traumatic surprise of diagnosis; (4) highs and lows of health care; and (5) support gap experienced postdiagnosis. Currently, there is no readily accessible psychosocial/educational information source for these patients. It is likely that a telephone intervention would be the most effective solution. PMID- 18389434 TI - "Coming to grips" with chemotherapy-induced premature menopause. AB - Chemotherapy for early stage breast cancer has significantly improved survival outcomes but is associated with ovarian toxicity, resulting in early menopause for many premenopausal women. A qualitative study was conducted that generated a grounded theory explaining how women carried on with life in response to breast cancer and menopause. My purpose in this article is to describe three distinct types of responses from women in that study: making the best of it, struggling and barely noticing. The degree of menopausal symptom distress and perceived level of preparation for the menopause experience had the greatest influence on the type of response. PMID- 18389435 TI - A different type of medicine: women's experiences with ophthalmic diseases in rural and urban Tamil Nadu, India. AB - I conducted a study to understand how urban and rural women conceptualized eye diseases in Tamil Nadu state(1). I chose to examine eye diseases because ailments such as cataracts, glaucoma, refractive error, and diabetic retinopathy rank among the most serious diseases of Indian women. I collected observational, survey, and interview data to compare women from rural and urban areas. In line with my expectations, rural women were more likely to rely upon non-medical means of treatment than their urban counterparts. Contrary to expectations, however, I found that urban and rural Tamil women's overall dependence on nonmedical means was not excessive. Based on the study's findings, it appears that both urban and rural women express a marked preference for both traditional and modern medical treatments - a phenomenon that is known as medical pluralism. Rural participants expressed a greater interest in medical pluralism than their urban counterparts. The results of the study lead me to believe that medical pluralism may be associated with the patient's geographic residence. I conclude this article with a discussion of the consequences of medical pluralism for women's health and overall societal development in Tamil Nadu. PMID- 18389436 TI - The meaning of women's experiences of living with multiple sclerosis. AB - We conducted a qualitative inquiry in order to describe the meaning of women's experiences of living with multiple sclerosis (MS). Multiple sclerosis is a chronic autoimmune disease of the central nervous system. The majority of persons living with MS are women. Living with MS has been described as difficult because of the uncertainty of the illness. Ten women with MS were interviewed and the interviews were analyzed with a phenomenological hermeneutic interpretation. In this study, we suggest that the meaning of living with MS for women can be understood as trying to maintain power and living with an unrecognizable body. The bodies of women with MS serve as hindrances in everyday life. Bodily changes evident to others impose feelings of being met in a different way, which can be understood as an expression of a violated dignity. At the same time, the women with MS struggle to protect their dignity. PMID- 18389437 TI - Unwrapping the box: dancing with the clients. AB - The authors describe the case of a girl suffering from anorexia nervosa who was regarded as a great challenge by her family therapist in Hong Kong. The metaphor of unwrapping a box was adopted to describe the therapeutic strategy of helping the girl express her inner feelings, and of helping the parents listen to their daughter describe her pain and suffering in the presence of the therapist. Health care professionals have to lead the dance, creating a space where their clients can share their secrets with their family and their therapist. PMID- 18389438 TI - In vitro effect of lycopene on cytokine production by human peripheral blood mononuclear cells. AB - There is evidence indicating that regular consumption of tomato products is associated with favorable immunomodulatory effects. In addition, tomato extracts have been shown to possess antioxidant, anticarcinogenic and antithrombotic activity in vitro. Since tomatoes are rich in carotenoids and particularly in lycopene--the pigment responsible for the red color of tomatoes--the present work was designed to examine the in vitro effect of lycopene on cytokine production by peripheral blood mononuclear cells (PBMC) from 15 healthy subjects. First, 2 x 10(6) PBMC suspended in 1 ml of conditioned medium were incubated over a period of 24 and 48 hours without or with the following concentrations of lycopene: 0.25, 0.5, 1.0, 2.0 and 4.0 microM. The production of the subsequent cytokines was evaluated: IL-1beta, IL-1ra, IL-2, IL-6 and IL-10, as well as TNFalpha and IFNgamma. Lycopene induced a dose-dependent increase in IL1beta, and TNFalpha production and a decrease in IL-2, IL-10 and IFNgamma secretion, whereas that of IL-6 and IL-1ra was not affected. It is concluded that understanding the role of lycopene in modulation of the immune system may promote decisions as for dietary supplementation of lycopene for reducing the risk of certain diseases. PMID- 18389439 TI - Effects of combined BCG and DHEA treatment in preventing the development of asthma. AB - Both BCG and dehydroepiandrosterone (DHEA) induce Th1 immune responses and suppress Th2 allergic reactions. To investigate whether the combination of BCG and DHEA has an additive effect on asthma prevention, BALB/c mice (n = 10 per group) were given an intraperitoneal injection of BCG at the beginning of sensitization, and fed mice chow containing DHEA throughout the study. In female mice, the combined administration of 2 x 10(4) CFUs BCG and 0.01% DHEA effectively suppressed the ovalbumin-induced increase in airway sensitivity to methacholine (56.5 vs. 8.2 mg/mL, p < 0.01), while BCG (13.9 mg/mL) or DHEA (17.9 mg/mL) alone did not. However, the addition of high dose (0.1%) DHEA decreased the efficacy of high dose (2 x 10(5) CFUs) BCG in suppressing the airway responsiveness and eosinophilia. In male mice, the treatments with BCG and/or DHEA were less effective, and the interferon-gamma/interleukin-4 ratio in the splenocyte supernatant was significantly higher and the ovalbumin-specific IgE concentration in the serum was significantly lower as compared to female mice. In conclusion, the combination of low doses of BCG and DHEA had an additive effect in suppressing the development of airway hypersensitivity. Androgens in males and DHEA overdose might reduce the efficacy of BCG. PMID- 18389440 TI - Autoimmune regulator initiates the expression of promiscuous genes in thymic epithelial cells. AB - The expression of peripheral antigens in the thymus, known as promiscuous gene expression, has been implicated in T cell tolerance and autoimmunity. Here we identified thymic epithelial cells (TECs) as the main cell type that expresses a diverse range of tissue-restricted antigens (TRAs). The TECs of a common autoimmune (non-obese diabetic [NOD]) mouse model express much lower levels of an autoimmune regulator (Aire) and TRAs than normal (Balb/c) TECs. Transfection of an Aire plasmid led to increased levels of TRA expression in cultured TECs from Balb/c and NOD mice; an increase that was enhanced by the presence of thymocytes. These data show that Aire initiates promiscuous gene expression in TECs, and that this function might be under thymocyte control. PMID- 18389441 TI - MHC-unrestricted lysis of MUC1-expressing cells by human peripheral blood mononuclear cells. AB - Many human adenocarcinomas can be killed in vitro by targeted cytotoxic T lymphocytes (CTL); however, major histocompatibility complex (MHC)-restrictions are typically required. The MUC1 antigen is common in many human adenocarcinomas, and is associated with a variable number of tandem repeats. It has been proposed that antigens with such repeated epitopes may be vulnerable to cytotoxic T lymphocyte killing without MHC-restriction. Therefore, it is possible that MUC1 expressing malignant cells may be killed by targeted cytotoxic T-lymphocyte in the absence of MHC-restriction. In this study, a human MUC1-expressing murine mammary carcinoma cell line was used to determine if cytotoxic T-lymphocyte killing of MUC1-expressing adenocarcinoma cells requires MHC-restriction. Specifically, MUC1-stimulated human mononuclear cells (M1SMC) were observed to kill human MUC1-transfected, MUC1-expressing murine mammary carcinoma cells, but not the mock-transfected, non-MUC1-expressing murine mammary carcinoma cells. Furthermore, the killing was blocked by antibody to MUC1, indicating MUC1 specific killing. In conclusion, cytotoxic T-lymphocyte killing of MUC1 expressing adenocarcinoma cells can be MHC-unrestricted. PMID- 18389442 TI - Estrogen modulates bone marrow-derived DCs in SLE murine model-(NZB x NZW) F1 female mice. AB - Dendritic cells (DCs) in the patient and animal models of systemic lupus erythematosus (SLE) are abnormal, but the detailed mechanism is unclear. Estrogen can modulate DCs in biological condition and estrogen concentration is related to the onset and development of SLE. So the control of estrogen on DCs might lead to the disorder of DCs. To prove the hypothesis, we detected the effects of 17beta estradiol (E2) on bone marrow (BM)-derived DCs in SLE murine model-(NZB x NZW) F1 (NZB/w F1) female mice before and after the disease onset. We found that E2 mainly enhanced the expression of surface molecule CD40, MHCII and the stimulation activity of immature DCs, but weakened the activity of mature DCs. E2 decreased the production of cytokines IL-6, IL-10, IL-12 and TNFalpha of DCs in young mice, but increased them in old mice. Tamoxifen could antagonize the E2 effect. E2 changed the expression of estrogen receptor-alpha (ER alpha) in DCs. The level of ER alpha in DCs of various old mice and the differentiation states varied. The results suggest that E2 can modulate the functions of BM-derived DCs in SLE pathology. The modulation is achieved by binding ER. The effects of E2 on DCs are different depending on the progression of SLE and cell differentiation status. This might be due to the difference of ER expression. PMID- 18389443 TI - The deposition of C5b-9 complexes and its precursors on E. coli J5 during complement activation enhances uptake and toxicities of gentamicin. AB - The complement system provides the host with protection against pathogenic agents and in some cases can result in damage to host tissue. However, the exact mechanism of how complement kills gram-negative bacteria in lysozyme-neutralized and or lysozyme-depleted serum is still under active investigation. In previous studies, it has been demonstrated that inner membrane damage by the membrane attack complex contributes to depolarization and the subsequent collapse of the membrane potential. In these studies we have shown that the membrane attack complex and its precursors provide additional protective effect by the enhanced uptake of antibiotics in the death of E. coli J5. Specifically, the deposition of C5b fragments from C6 neutralized Pooled Normal Human Serum (PNHS) and C5b6 complexes from C7 neutralized PNHS on E. coli J5 contribute to antibiotic uptake and killing. Since C5b and C5b6 do not form pores, we suggest that disturbances and or cracks in the outer membrane by the deposited complexes accelerates uptake of the antibiotics and enhanced killing of E. coli J5 employed in these studies. PMID- 18389444 TI - Detecting esophageal disease with second-generation capsule endoscopy: initial evaluation of the PillCam ESO 2. AB - BACKGROUND AND STUDY AIM: Esophageal capsule endoscopy (ECE) provides an alternative, minimally invasive modality for evaluating the esophagus. This study evaluates the performance and test characteristics of a second-generation esophageal capsule endoscope, the PillCam ESO 2. METHODS: Adults with known or suspected esophageal disease were included. Using the simplified ingestion procedure, each patient underwent capsule endoscopy with the PillCam ESO 2. Following ECE, esophagogastroduodenoscopy (EGD) was performed on the same day by an investigator who was blinded to the results of the ECE. In random order, capsule endoscopy videos were read and interpreted by the study investigator blinded to EGD results. RESULTS: 28 patients (19 men, 9 women; mean age 53.3 years) were included. In 82 % of the patients, at least 75 % of the Z line was visualized by the PillCam ESO 2. A per-lesion analysis demonstrated that the PillCam ESO 2 had definitive results in 30/43 lesions (69.8 %) and EGD in 29/43 (67.4 %), P value = 0.41. Compared with EGD for detecting suspected Barrett's esophagus and esophagitis, the PillCam ESO 2 had a sensitivity of 100 % and a specificity of 74 %, and a sensitivity of 80 % and a specificity of 87 %, respectively. The PillCam ESO 2 demonstrated 86 % agreement with EGD in describing the Z line (kappa statistic 0.68). The modified ingestion protocol provided excellent cleansing, with bubbles/saliva having no or only a minor effect on Z line images in 86 % of cases. CONCLUSIONS: The PillCam ESO 2 demonstrated excellent visualization of the Z line. Compared with standard EGD, the PillCam ESO 2 had good test characteristics with high rates of detection of suspected Barrett's esophagus and esophagitis. This study provides indirect validation of the simplified ingestion procedure. The PillCam ESO 2 acquires high quality esophageal images, performs safely, and should be able to replace the current PillCam ESO. PMID- 18389445 TI - Endoscopic mucosal resection for early gastric cancer: comparison of two modifications of the cap method. AB - BACKGROUND AND STUDY AIM: Endoscopic mucosal resection using a cap (EMR-C) is an established method for curative resection of early neoplastic lesions; prelooping of the snare may however be difficult and lead to imprecise resection. We therefore compared two modifications of the conventional technique using outer snare placement with an accessory channel in a prospective, nonrandomized study. PATIENTS AND METHODS: Between October 2004 and March 2007, 54 patients (men 37, women 17; mean age 71 years) underwent EMR. One method involved an internally retained snare (IRS) cap, with a fixed prelooped snare inside the cap; the other method used an externally guided snare (EGS) cap with the snare guided over an oblique cap. The main outcome parameters were specimen size, en bloc resection, and complications. RESULTS: There was no difference between use of the IRS and EGS cap methods in relation to specimen size (27.6 vs. 27.1 mm), or rates of en bloc resection (88.9 % vs. 83.3 %); only one perforation occurred, and this was in the EGS group. CONCLUSION: Both techniques appeared to provide similar efficacy, the inner rim of the IRS cap stabilizes aspiration of the lesion compared with the EGS cap that does not have it. PMID- 18389446 TI - Miss rate for colorectal neoplastic polyps: a prospective multicenter study of back-to-back video colonoscopies. AB - BACKGROUND AND STUDY AIM: Polyp miss rates during colonoscopy have been calculated in a few tandem or back-to-back colonoscopy studies. Our objective was to assess the adenoma miss rate while limiting technique or operator expertise biases, i. e. by performing a large multicenter study, with same-day back-to-back video colonoscopy, done by two different operators in randomized order and blinded to the other examination. PATIENTS AND METHODS: 294 patients at 11 centers were included. Among the 286 analyzable tandem colonoscopies, miss rates were calculated in both a lesion- and patient-based analysis. Each of these rates was determined for polyps overall, for adenomas, and then for lesions larger than 5 mm, and for advanced adenomas. Univariate and logistic regression analysis were performed to define independent variables associated with missed polyps or adenomas. RESULTS: The miss rates for polyps, adenomas, polyps > or = 5 mm, adenomas > or = 5 mm, and advanced adenomas were, respectively, 28 %, 20 %, 12 %, 9 % and 11 %. None of the masses with a carcinomatous (n = 3) or carcinoid component (n = 1) was missed. The specific lesion miss rates for patients with polyps and adenomas were respectively 36 % and 26 % but the corresponding rates were 23 % and 9.4 % when calculated for all 286 patients. The diameter (1-mm increments) and number of polyps (> or = 3) were independently associated with a lower polyp miss rate, whereas sessile or flat shape and left location were significantly associated with a higher miss rate. Adequacy of cleansing, presence of diverticula, and duration of withdrawal for the first procedure were not associated with adenoma miss rate. CONCLUSIONS: We confirm a significant miss rate for polyps or adenoma during colonoscopy. Detection of flat polyps is an issue that must be focused on to improve the quality of colonoscopy. PMID- 18389447 TI - Prospective audit of colonoscopy quality in Kent and Medway, UK. AB - AIM: To compare the quality of colonoscopy in the Kent and Medway Strategic Health Authority with national standards and previous audits. METHOD: A prospective 12-month audit of colonoscopy quality as assessed by number of procedures performed, total colonoscopy rates, sedation usage, and complications. Data were collected by 7 endoscopy units on 5905 colonoscopies performed by 62 colonoscopists. The endoscopy unit nurses, as opposed to the colonoscopists, verified that colonoscopy was total. RESULTS: Seven doctors stopped performing colonoscopy during the study period. Thirty-nine of 55 colonoscopists (71 %) achieved total colonoscopy in at least 90 % of cases; 12 (22 %) completed colonoscopy in 80 - 89 % of their cases and 4 (7 %) in 79 % or less of their cases. Seventy-nine percent of colonoscopists used sedation in accordance with British Society of Gastroenterology (BSG) guidelines. Only 22 of 55 (40 %) of colonoscopists performed more than 100 colonoscopies during the 12-month audit period. Reported complications were below expected levels. CONCLUSION: In our study almost one-third of colonoscopists did not achieve colonoscopy completion rates of at least 90%, and less than half performed more than 100 colonoscopies during the 12 month study. Adherence to quality standards appears to be inadequate. PMID- 18389448 TI - A prospective randomized trial of cannulation technique in ERCP: effects on technical success and post-ERCP pancreatitis. AB - BACKGROUND AND STUDY AIMS: Inadvertent injection of contrast agent into the pancreatic duct is believed to be an important contributor to pancreatitis occurring after endoscopic retrograde cholangiopancreatography (post-ERCP pancreatitis, PEP). Our aim was to examine whether primary deep biliary cannulation with a guide wire is associated with a lower rate of PEP than conventional contrast-assisted cannulation. PATIENTS AND METHODS: From August 2003 to April 2006 all patients with an intact papilla who were referred for ERCP were eligible. Patients with pancreatic or ampullary cancer were excluded. Patients were randomized to undergo sphincterotomy biliary cannulation using either contrast injection or a guide wire. The ERCP fellow attempted initially for 5 minutes. If unsuccessful, the consultant attempted for 5 minutes using the same technique, followed by crossover to the other technique in the same sequence and then needle-knife sphincterotomy where appropriate. Patients were assessed clinically after the procedure, then followed up with telephone interviews after 24 hours and 30 days, and serum amylase and lipase tests after 24 hours. RESULTS: Out of 1654 patients undergoing ERCP, 413 were included in the study. PEP occurred in 29/413 (7.0 %): 16 in the guide-wire arm, 13 in the contrast arm ( P = 0.48). The overall cannulation success rate was 97.3 %. Cannulation was successful without crossover in 323/413 patients (78.2 %): 167/202 (81.4 %) in the guide-wire arm and 156/211 (73.9 %) in the contrast arm ( P = 0.03). Multivariate analysis demonstrated female sex (OR = 2.7, P = 0.04), suspected sphincter of Oddi dysfunction (OR = 5.5, P = 0.01), and complete filling of the pancreatic duct with contrast agent (OR = 3.5, P = 0.02) to be independently associated with PEP. The risk of PEP increased incrementally with each attempt at the papilla (OR 1.4 per attempt, P = 0.04) to greater than 10 % after four or more attempts. CONCLUSIONS: The guide-wire technique improves the primary success rate for biliary cannulation during ERCP but does not reduce the incidence of PEP compared to the conventional contrast technique. The incidence of PEP increases incrementally with each attempt at the papilla. PMID- 18389449 TI - Endoscopic ultrasound-guided application of a new hybrid cryotherm probe in porcine pancreas: a preliminary study. AB - BACKGROUND AND STUDY AIMS: Open, laparoscopic, or percutaneous radiofrequency (RF) ablation of the pancreas is still dangerous, whereas endoscopic ultrasound (EUS)-guided ablation might reduce risk because it is less invasive and provides real-time monitoring. We aimed to demonstrate the feasibility of transluminal RF ablation and to evaluate the efficacy and safety of a new flexible bipolar ablation probe combining RF and cryotechnology. METHODS: 14 ablations were performed in 14 pigs. Energy input (16 W) and simultaneous cryogenic cooling with carbon dioxide (650 psi) were standardized. Application time range was 120 - 900 seconds. Ablation area was measured by EUS immediately after ablation (area T0), and before euthanasia (area T1). Macroscopic findings (area T2) and histological findings after necropsy served as gold standard. The interval from application to euthanasia was either 1 or 2 weeks. RESULTS: The correlation between EUS findings (area T1) and macroscopic appearance (area T2) was good ( R = 0.89). The correlation between the T2 ablation area and the application time showed a fitted ratio of 2.3 ( P < 0.0001) with a 1-week interval and 0.2 ( P = 0.01) with a 2 week interval. No pig died because of the procedure. Two pigs showed histochemical pancreatitis, which was clinically overt in one. Necropsy additionally revealed one burn to the gastric wall and four gut adhesions. CONCLUSIONS: Selective transluminal RF ablation of the pancreas under EUS control in a living pig model is feasible. The new flexible bipolar probe creates an ablation area with extent related to the duration of application, and with fewer complications than conventional RF ablation techniques. PMID- 18389450 TI - Detection of neoplasia at colonoscopy: what next? PMID- 18389451 TI - EUS-guided choledochoduodenostomy for palliative biliary drainage in patients with malignant biliary obstruction: results of long-term follow-up. AB - Five patients with obstructive jaundice caused by malignant periampullary biliary stenosis underwent EUS-guided choledochoduodenostomy (EUS-CDS) from the first portion of the duodenum using a convex echoendoscope and a needle knife. All the steps of the procedure including passage dilatation and the plastic stent placement were performed through the accessory channel of the echoendoscope over the guide wire. Stent insertion was technically successful in all five patients. The procedure was also clinically effective in relieving jaundice in all cases. One patient developed pneumoperitoneum, which resolved with conservative management. Stent exchange was successful in seven of eight attempts in patients with stent occlusion. One failure was due to tumor invasion to the choledochoduodenal fistula. Stent patency was maintained in the remaining patients throughout their survival period. The average stent patency was 211.8 days. EUS-CDS from the first portion of the duodenum appears to be feasible and safe in cases of obstructive jaundice caused by distal bile duct obstruction. PMID- 18389452 TI - Heller-type cardiomyotomy using NOTES. PMID- 18389454 TI - Emergence of "cat scratch colon" during a colonoscopy. PMID- 18389456 TI - [Moyamoya disease among children. An analysis of a series in the Western world and literature review]. AB - PATIENTS AND METHODS: A total of 12 patients with moyamoya disease or syndrome with a mean age of 6 years were analyzed in a retrospective fashion. RESULTS: Infarction was the most frequent presentation feature (8 out of 12 patients), 2 had epilepsy and 2 were incidental findings. Seven cases were classified as idiopathic while the other five were related to systemic illnesses. Diagnosis was initially made by magnetic resonance angiography in 9 cases, and conventional angiography in 3 cases. Mean follow-up is 5 years; six patients experienced clinical worsening of symptoms, while 6 cases remained clinically stable. However, all of them showed angiographic progression. Four patients underwent revascularization surgery. Two children died due to complications associated with moyamoya disease, and six have moderate handicaps. CONCLUSION: Due to the fact moyamoya disease is not an indolent disorder and readily progress to cause complications, surgical revascularization should always be considered in the management of these patients. PMID- 18389457 TI - [Endovascular treatment of carotid disease in patients at high surgical risk]. AB - AIMS: To analyse the perioperative morbi-mortality rates during the first 30 days following endovascular therapy in patients with carotid stenosis and important risk factors for surgery. We also sought to determine the survival rate, the percentage of patients that were stroke-free and the percentage of cases of restenosis during the follow-up period. PATIENTS AND METHODS: A retrospective study was conducted on a series of 100 patients with symptomatic > 50% and asymptomatic > 70% carotid stenosis, who underwent endovascular therapy between January 2000 and December 2006 because of the important risk factors they presented. Monitoring was clinical and was performed by means of carotid Doppler scanning. RESULTS: The mean age of the sample: 72.2 years (46-86). Perioperative morbi-mortality of the series: 6% (confidence interval, CI 95% = 0.8-11.2) and in the symptomatic stenosis group: 5.7% (CI 95% = 1.8-12.9). The mean follow-up time was 23.4 months (0-94). Total mortality of the series during the follow-up was 7% (CI 95% = 1.4-12.5). The probability of survival at 3 and 5 years is 93 and 89%, respectively. At one year, 98.9% of the patients remained stroke-free. Restenosis of the stent occurred in 5% (CI 95% = 1.6-11.2). The probability of restenosis not occurring was 96.75% at 6 months and 94% at 3 years. CONCLUSIONS: Stent angioplasty is an effective form of treatment in carotid stenosis in patients with important risk factors for surgery and it is therefore important to fulfil patient selection protocols and avoid perioperative complications. PMID- 18389458 TI - [Estimation of the prevalence, incidence, comorbidities and direct costs associated to stroke patients requiring care in an area of the Spanish population]. AB - AIM: To determinate the prevalence, incidence, co-morbidities and therapeutic objectives and costs of stroke among Spanish population. PATIENTS AND METHODS: A retrospective study was performed based on data from patients attended for stroke, aged > 30 years, from five Spanish primary care centres and two hospitals in 2006. Comparative group: patients without stroke. Main analysed variables were: age, sex, co-morbidity (cardiovascular/others), clinical parameters and direct costs (pharmacy, derivations, visits, emergencies, procurement, and hospitalisation). An ANCOVA analysis and logistic regression were used to fit the model. RESULTS: A 4.5% of 57.026 patients (n = 2.585; CI 95% = 4.3-4.7%) suffered stroke. The incidence of stroke was 220 new-cases/100.000 populations. Main differences between patients suffering stroke/control group were: age (72.5 vs. 53.5), men (58.2% vs. 44.6%), episodes/year (7,9 vs. 4,8), visits/year (15,8 vs. 8,1), p < 0,001. Stroke had an independent relation with age (OR = 1,4), male (OR = 2,3), diabetes (OR = 1,6), hypertension (OR = 1,5), smoking (OR = 1,5), alcohol (OR = 1,4), depression (OR = 1,4), dyslipidemia (OR = 1,3) and dementia (OR = 1,2). Some of the results were: systolic pressure (134.1 vs. 127.6 mmHg) and LDL cholesterol (116.4 vs. 126.2 mg/dL), in presence/absence of stroke, p < 0,001. The average of annual costs of stroke was 2,590.36 vs. 985.26 euros, p < 0.001. After the correction of the logistic model results did not change: 1,774.33 (CI 95% = 1,720.10-1.828.55) vs. 1,021.98 euros (CI 95% = 1,010.92-1,033.03), p < 0,001. All components of costs were higher in the stroke group. CONCLUSIONS: Patients that demanded assistance for stroke had a higher number of co morbidities and a higher total cost/patient/year. Therapeutic objectives could be improved, mainly in primary prevention of cardiovascular risk factors. PMID- 18389459 TI - [Gait analysis and quality of life evaluation after gait training in patients with spinal cord injury]. AB - INTRODUCTION: Over recent years it has become possible to retrain motor activity among patients with partial spinal cord injury, especially for walking. AIM. To assess the impact of gait training on a treadmill with body weight support, regarding temporospatial parameters and quality of life. PATIENTS AND METHODS: Twelve patients of both sexes were evaluated. They had been diagnosed with partial spinal cord injury of traumatic origin at least 12 months earlier. They were able to walk and their motor function below the level of the injury was partially preserved and classified as level C or D. After the initial evaluation, the participants were trained on a treadmill with body weight support, with two sessions per week lasting 30 minutes each, over a four-month period, thus totaling 30 sessions. The patients' temporospatial gait parameters and quality of life were analyzed before and after the training. RESULTS: There were improvements in all the temporospatial parameters evaluated (p < 0.0001), but no changes in quality of life were seen (p > 0.05). CONCLUSION: Treadmill training with body weight support among patients with spinal cord injury was effective in improving the temporospatial gait parameters, but without changing their quality of life. PMID- 18389460 TI - [Simultaneous suprasellar and pineal germinoma: a case report]. AB - INTRODUCTION: Tumours in the pineal region are rare (0.3-2.7%) and most of the ones that do occur are germ cell tumours, of which germinoma is the most frequent. They are chiefly located in the pineal gland, the next most common being site being the suprasellar region and the fourth ventricle, although simultaneous presentation in these locations is relatively rare (5-10% of cases). The main aim of this study is to review the differential diagnosis of neoplasias in the pineal region. CASE REPORT: Here we report the case of a 20-year-old male who was admitted to hospital due to irregularities affecting the field of vision, with clinical signs and symptoms of panhypopituitarism and normal levels of tumour markers in blood and cerebrospinal fluid. Magnetic resonance imaging of the craniospinal axis was performed and results showed the presence of two masses with significant enhancement, one located in the pineal region with a signal similar to that of grey matter and the other in the hypothalamus, which had a more heterogeneous aspect with cystic areas inside it. Imaging with paramagnetic contrast agents revealed dissemination with ependymal uptake in relation to the frontal horns and in the fourth ventricle; a possible simultaneous origin was also suggested in this latter location. CONCLUSIONS: Magnetic resonance imaging plays a decisive role in establishing the diagnosis and therapy plan. The imaging factors that must be taken into account are the origin of the mass, heterogeneity, contrast enhancement, presence of fat or calcium, a signal similar to that of grey matter and possible dissemination, above all into the subarachnoid space. The patient's medical record (age, sex or tumour markers) must also be taken into consideration. PMID- 18389461 TI - [The NEDICES cohort of the elderly. Methodology and main neurological findings]. AB - AIM: To summarise the methodological aspects and main findings of the NEDICES (Neurological Disorders in Central Spain) cohort study of the elderly population, with over 5000 participants. DEVELOPMENT: The article outlines the findings concerning the following chronic neurological diseases (CND): dementia, Parkinson's disease and Parkinsonisms, stroke and essential, or senile, tremor. The NEDICES study examined the health, mortality and a series of sociological aspects of the cohort that are not discussed here. The paper does describe, however, the objectives, methods, population and development, with the baseline (1994) and incidence (1997) cut-off points, and the main findings regarding the CND under study are discussed. CONCLUSIONS: The prevalence and incidence of the CND are comparable to those of other Spanish and European population-based studies, but with certain peculiarities, such as the fact that incidence of senile tremor is the highest of those reported in the literature and that this CND is associated with dementia. Over half the cases of Parkinson's disease were reported de novo with the study and Alzheimer's disease was associated with vascular risk factors. PMID- 18389462 TI - [The physiopathology and management of sexual dysfunction in epileptic patients]. AB - AIM: To review sexual dysfunction in epileptic patients, which is an aspect of the disease that is often deemed as being of little importance, but which exerts a decisive influence on the quality of life of these patients. DEVELOPMENT: The alterations in sexual functioning in epilepsy have a complex physiopathology, can be of different types and occur during seizures, the aura or in the intercritical periods. Their clinical expression depends on the sex and age of the patient. The severity and incidence of sexual dysfunction are influenced by the treatment being used, the psychosocial adjustment of the individual, and certain aspects of epilepsy such as the age at onset, time to progression, location of the focus, the type of seizures and the degree of control the patient has over them. The therapeutic strategy is based on adjusting or modifying the antiepileptic therapy and then treating the sexual dysfunction and/or possible hormonal upsets. CONCLUSIONS: Sexuality is a fundamental human right and as a health provider, the physician must take this aspect of the disease into account when dealing with epileptic patients by attempting to detect and characterise the disorder. The patient must be informed of the possible effects of epilepsy and its treatment on sexual functioning, and also the therapeutic options the physician considers to be best suited to the patient's particular case. He or she should also be encouraged to play an active role in making decisions on the matter and it is necessary to carry out developmental monitoring that takes into account the impact that improved sexual functioning is going to have on the patient's psychosocial and family adjustment. PMID- 18389463 TI - [Functional neuroimaging with SPECT in Parkinson's disease and Parkinsonisms]. AB - INTRODUCTION: The diagnosis of Parkinson's disease and the different Parkinsonian syndromes is currently determined by applying established clinical criteria. Today, different neuroimaging techniques, such as positron emission tomography and single-photon emission computerised tomography (SPECT), are a valuable aid to further our understanding of both the pathophysiology and the diagnosis of the different Parkinsonian pictures. AIM. To examine the usefulness of the different tools utilised to assess the integrity of the nigrostriatal system by visualising the pre-synaptic dopamine transporters and post-synaptic D2 receptors using SPECT techniques; we also sought to determine the role played by the alteration of the cardiac sympathetic innervation using meta-iodobenzylguanidine. DEVELOPMENT: We review the usefulness of the different nuclear medicine techniques for the diagnosis and differential diagnosis of the distinct clinical pictures that are accompanied by Parkinsonian symptoms, and we also discuss the possible future perspectives of said techniques. CONCLUSIONS: A combination of the different nuclear medicine techniques that have been reviewed here is useful in the study and characterisation of Parkinsonian syndromes. In the future they could be used to detect subjects who are possibly at risk of developing Parkinsonism and to be able to assess the effect of therapies that may have a potential modifying effect on the disease. PMID- 18389465 TI - [Menkes disease: its clinical, biochemical and molecular diagnosis]. PMID- 18389466 TI - [Lacunar stroke as the presenting symptom of Crohn's disease]. PMID- 18389467 TI - [Carotid dissection in a cocaine-abusing patient]. PMID- 18389468 TI - A potent tyrosinase activator from Radix Polygoni multiflori and its melanogenesis stimulatory effect in B16 melanoma cells. AB - Tyrosinase is a key enzyme in melanin biosynthesis. Activators of tyrosinase with stimulatory effects on melanogenesis are beneficial for the treatment of hypopigmentation diseases. In the present study, mushroom tyrosinase activity assay was performed to screen tyrosinase activators from traditional Chinese herbs. Four components from Radix Polygoni multiflori were tested. The most active compound, 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside (THSG), was found to be a significant tyrosinase activator. The maximal activation was 126% at a concentration of 75.0 microg/mL. The three anthraquinones slightly activated tyrosinase with effects in the range 7-31%. All the compounds were tested in B16 melanoma cells, the anthraquinones were found to inhibit cell proliferation at a concentration of 0.1-2.5 microg/mL, and THSG was found to be non-cytotoxic at a concentration of 0.1-12.5 microg/mL. THSG significantly increased the activity of murine tyrosinase and stimulated melanin biosynthesis in B16 melanoma cells. In conclusion, THSG is a potent tyrosinase activator and stimulator of melanogenesis with potential for the treatment of hypopigmentation disease. PMID- 18389469 TI - Isolation and identification of cytotoxic compounds from the wood of Pinus resinosa. AB - Methanol extracts of wood from Pinus resinosa were found to be selectively cytotoxic against human lung carcinoma cells, A549 (IC50 41 +/- 6 microg/mL), human colorectal adenocarcinoma cells, DLD-1 (IC50 47 +/- 4 microg/mL) in comparison with healthy cells, WS1 (IC50 130 +/- 11 microg/mL). Five known compounds were isolated and identified by 1H, 13C NMRspectroscopy and HR-ESI-MS mass spectrometry as, pinosylvin monomethyl ether (1), pinosylvin (2), pinosylvin dimethyl ether (3), pinobanksin (4) and (-)-norachelogenin (5). Compound 4 was isolated for the first time in P. resinosa. The cytotoxicity of compounds 1-5 was evaluated against A549, DLD-1 and WS1. Compound 1 exhibited the strongest cytotoxicity against both tumor cell lines and the healthy cell line with an IC50 of 25 +/- 4 microm for A549, 20 +/- 1 microm for DLD-1 and 34 +/- 3 microm for WS1. PMID- 18389470 TI - Evaluation of antioxidant and antimicrobial properties of Soymida febrifuga leaf extracts. AB - The present study was designed to evaluate the antioxidant and antimicrobial properties of hexane (LH), methanol (LM) and aqueous (LA) extracts of Soymida febrifuga (Maliaceae) leaves, which is a traditional folk medicine in India. No pharmacological evaluation of this plant (except antiplasmodial activity) has been reported to date. Antioxidant activity of different extracts was evaluated by DPPH free radical scavenging activity, taking total phenolic content (TPC) as an index. Antimicrobial activity was tested against six bacterial and five fungal strains using the agar hole diffusion method and the minimum inhibitory concentrations (MIC) and minimum microbicidal concentration (MMC) were determined for all the test organisms against the extracts. The results showed that the methanol and aqueous extracts of leaf had a higher antioxidant activity and total phenolic content than the hexane extract. The antioxidant activity and TPC of the extracts were highly correlated. Extracts also showed several degrees of antimicrobial activity against different microbes. The methanol extract was more potent against Aspergillus fumigatus and Candida tropicana. The lowest MIC values obtained for LM, LA and LH were 78, 156, 625 microg/mL against A. fumigatus, C. tropicana and C. albicans, respectively. Hence, this study confirms that Soymida febrifuga leaves possess potent antioxidant and antimicrobial activity. PMID- 18389471 TI - Isolation of linoleic and alpha-linolenic acids as COX-1 and -2 inhibitors in rose hip. AB - Rose hip has previously shown clinical efficacy in the treatment of osteoarthritis, and organic solvent extracts of rose hip have showed inhibition of cyclooxygenase-1 and -2. A petroleum ether extract of rose hip was fractioned by VLC on silica; on a C-18 column and by HPLC. Each step was COX-1/2 activity guided. The bioassay-guided fractionation led to the isolation of linoleic acid (the IC50 for COX-1 was 85 microm and 0.6 microM for COX-2) and alpha-linolenic acid (the IC50 for COX-1 was 52 microM and 12 microM for COX-2). The COX-2/COX-1 ratio was 0.007 for linoleic acid and 0.2 for alpha-linolenic acid. Linoleic acid and alpha-linolenic acid contribute to the COX-1 and -2 inhibitory activity of rose hip. PMID- 18389472 TI - Antimicrobial activity of various parts of Polyalthia longifolia var. pendula: isolation of active principles from the leaves and the berries. AB - Methanol extracts of leaves, stem, twigs, green berries, flowers, roots, root wood and root-bark of Polyalthia longifolia var. pendula, were tested for their antibacterial and antifungal potentials. Bioassay monitored isolation work on the methanol extract of leaves and berries which possess promising antibacterial activity led to the isolation of seven clerodane diterpenoids, 16(R and S) hydroxy-cleroda-3,13(14)Z-dien-15,16-olide (1), 16-oxo-cleroda-3,13(14)E-dien-15 oic acid (2), methyl-16-oxo-cleroda-3,13(14)E-dien-15-oate (3), 2-oxo-kolavenic acid (4), 16 (R and S)-hydroxy-cleroda-3,13(14)Z-dien-15,16-olide-2-one (5), (4- >2)-abeo-16(R and S)-hydroxy-cleroda-2,13(14)Z-dien-15,16-olide-3-al (6), 3beta,16alpha-dihydroxy-cleroda-4(18), 13(14)Z-dien-15,16-olide (7), while kolavenic acid (8) and solidagonal acid (9) were obtained from the root-wood. Diterpenoids 1 and 8 were also obtained from the root-bark. It is the first report of the isolation of 7 and 9 from this source, and clerodane 3 was obtained as a natural product for the first time. Clerodanes 1, 2, 5, 6 and 7 were found to be active antimicrobial agents with MIC values ranging between 7.8 and 500 microg/mL. Diterpenoid 1 emerged as the most active antimicrobial agent. The acetyl derivative (10) of 1 and the methyl derivative (3) of 2 were found to be less active than the parent compounds. A complex of allantoin was also obtained from the berries, which on hydrolysis furnished pure allantoin (11). PMID- 18389473 TI - Effect of SBD.4A--a defined multicomponent preparation of Angelica sinensis--in periodontal regeneration models. AB - Periodontitis is a major cause of tooth motility and loss, resulting in destruction of the supporting structures of the tooth, including periodontal ligaments and alveolar bone. Periodontal surgery can slow the progression of the disease, but is costly, invasive, limited by contraindications and technique sensitive. Recently, non-invasive pharmacological treatments using proteinaceous biologicals have become available. Here, for the first time, the bone regenerative capabilities of a non-proteinaceous biological--SBD.4A--a novel, stable multicomponent growth factor isolated from a medicinal plant Angelica sinensis are reported. SBD.4A was tested in osteoblast proliferation and differentiation systems, as well as in a fibroblast-secreted hyaluronic acid assay. Furthermore, SBD.4A was formulated in a slow release matrix and tested in the rat calvarial defect model. Apart from the previously reported strong stimulation of angiogenesis, fibroblast growth and collagen synthesis--the activities needed for periodontal regeneration--SBD.4A enhanced the deposition of hyaluronic acid and proliferation of osteoblasts in vitro, as well as bone regeneration in the rat calvarial defect model. Together, these results indicate the beneficial effect of SBD.4 on periodontal ligament and bone regeneration making the case for further development of this botanical growth factor. PMID- 18389474 TI - Pharmacological and immunological effects of individual herbs in the Food Allergy Herbal Formula-2 (FAHF-2) on peanut allergy. AB - It was previously shown that a Chinese herbal formula, Food Allergy Herbal Formula 2 (FAHF-2) composed of nine herbs, blocked peanut-induced anaphylaxis in a murine model. The current study was designed to investigate the pharmacological actions of individual herbs comprising FAHF-2 on peanut-induced anaphylactic reactions in a murine model of peanut allergy and to determine if all nine herbs are necessary to prevent an anaphylactic reaction, or if a simplified formula containing fewer herbs would be equally effective. Some individual herbs reduced peanut-induced anaphylactic symptoms but no single herb offered full protection from anaphylactic symptoms equivalent to FAHF-2. The herbs had highly variable effects on histamine release, as well as peanut-specific serum IgE and IgG2a levels. The herbs also had variable effects on IL-4, IL-5 and IFN-gamma levels. A simplified formula comprising the most efficacious tested individual herbs showed only partial efficacy and was not able to reproduce comparably the effects of FAHF-2, suggesting that component herbs of FAHF-2 may work synergistically to produce the curative therapeutic effects produced by the whole formula, which appears to be the best option for future clinical trials. PMID- 18389475 TI - Activation drives PD-1 expression during vaccine-specific proliferation and following lentiviral infection in macaques. AB - Recent data supports that increased expression of PD-1, a negative regulator of immune function, is associated with T cell exhaustion during chronic viral infection. However, PD-1 expression during acute infection and vaccination has not been studied in great detail in primates. Here, we examine PD-1 expression on CD3(+) T cells following DNA vaccination or lentiviral infection of macaques. Ex vivo peptide stimulation of PBMC from DNA-vaccinated uninfected macaques revealed a temporal increase in PD-1 expression in proliferating antigen-specific CD8(+) T cells. Following the initial increase, PD-1 expression steadily declined as proliferation continued, with a concomitant increase in IFN-gamma secretion. Subsequent examination of PD-1 expression on T cells from uninfected and lentivirus-infected non-vaccinated macaques revealed a significant increase in PD 1 expression with lentiviral infection, consistent with previous reports. PD-1 expression was highest on cells with activated memory and effector phenotypes. Despite their decreased telomere length, PD-1(hi) T cell populations do not appear to have statistically significant uncapped telomeres, typically indicative of proliferative exhaustion, suggesting a different mechanistic regulation of proliferation by PD-1. Our data indicate that PD-1 expression is increased as a result of T cell activation during a primary immune response as well as during persistent immune activation in macaques. PMID- 18389476 TI - Human Thy-1 induces secretion of matrix metalloproteinase-9 and CXCL8 from human neutrophils. AB - Neutrophils are the first cells arriving at sites of acute inflammation. On their way from blood to the site of inflammation, neutrophils have to adhere to endothelial cells (EC), to transverse the basement membrane and subsequently to travel through the interstitial matrix. Recently, we have shown that human Thy-1 is an alternate EC receptor for the leukocyte integrin Mac-1 that contributes to leukocyte recruitment to sites of inflammation, providing a new pathway for adhesion and transmigration of neutrophils. Here, we studied the effect of Thy-1 mediated adhesion on neutrophil functions. Binding of neutrophils to recombinant human Thy-1 stimulated the release of MMP-9 from neutrophils, resulting in their enhanced migration through collagen-IV and matrigel. Further, we showed that neutrophil interaction with Thy-1 stimulated secretion of CXCL8 and thus could support the attraction of additional neutrophils to inflammatory sites. Blocking experiments confirmed the pivotal roles of Thy-1 on activated dermal EC or fibroblasts and its counter receptor CD18 on neutrophils for the regulation of MMP-9 and CXCL8 release from neutrophils. Our results support the general concept that the function of 'adhesion molecules' in particular of human Thy-1, may not only be to provide mechanical support but also regulate neutrophil functions. PMID- 18389478 TI - Nanoparticles target distinct dendritic cell populations according to their size. AB - The efficiency of a vaccine largely depends on the appropriate targeting of the innate immune system, mainly through prolonged delivery of antigens and immunomodulatory substances to professional antigen-presenting cells in the lymphoid environment. Particulate antigens, such as virus-like particles (VLP) induce potent immune responses. However, little is known about the relative importance of direct drainage of free antigen to lymph nodes (LN) versus cellular transport and the impact of particle size on the process. Here, we show that nanoparticles traffic to the draining LN in a size-dependent manner. Whereas large particles (500-2000 nm) were mostly associated with dendritic cells (DC) from the injection site, small (20-200 nm) nanoparticles and VLP (30 nm) were also found in LN-resident DC and macrophages, suggesting free drainage of these particles to the LN. In vivo imaging studies in mice conditionally depleted of DC confirmed the capacity of small but not large particles to drain freely to the LN and demonstrated that DC are strictly required for transport of large particles from the injection site to the LN. These data provide evidence that particle size determines the mechanism of trafficking to the LN and show that only small nanoparticles can specifically target LN-resident cells. PMID- 18389477 TI - Interleukin-10-induced MARCH1 mediates intracellular sequestration of MHC class II in monocytes. AB - IL-10 is a potent anti-inflammatory cytokine interfering with antigen presentation by inducing the intracellular sequestration of MHC class II (MHC-II) molecules. Here we studied the contribution of membrane-associated RING-CH (MARCH) ubiquitin ligase family members to the IL-10-induced down-regulation of MHC-II molecules. We found that MARCH1 and MARCH8 proteins are the most potent family members for the down-regulation of MHC-II surface expression in transfected cells, but only MARCH1 mRNA expression is strongly induced by IL-10 in human primary monocytes. We detected mono- and poly-ubiquitinated forms of MHC II molecules both in IL-10-treated monocytes and in cells transfected with MARCH1. We also show direct interaction between MHC-II and MARCH1 molecules in co immunoprecipitation assays. Finally, we found that siRNA-mediated knockdown of MARCH1 reverses IL-10-induced MHC-II down-regulation in primary monocytes. Thus, the immunosuppressive effect of IL-10 on antigen presentation is mediated through induced expression of MARCH1. PMID- 18389479 TI - DiC14-amidine cationic liposomes stimulate myeloid dendritic cells through Toll like receptor 4. AB - DiC14-amidine cationic liposomes were recently shown to promote Th1 responses when mixed with allergen. To further define the mode of action of diC14-amidine as potential vaccine adjuvant, we characterized its effects on mouse and human myeloid dendritic cells (DC). First, we observed that, as compared with two other cationic liposomes, only diC14-amidine liposomes induced the production of IL 12p40 and TNF-alpha by mouse bone marrow-derived DC. DiC14-amidine liposomes also activated human DC, as shown by synthesis of IL-12p40 and TNF-alpha, accumulation of IL-6, IFN-beta and CXCL10 mRNA, and up-regulation of membrane expression of CD80 and CD86. DC stimulation by diC14-amidine liposomes was associated with activation of NF-kappaB, ERK1/2, JNK and p38 MAP kinases. Finally, we demonstrated in mouse and human cells that diC14-amidine liposomes use Toll-like receptor 4 to elicit both MyD88-dependent and Toll/IL-1R-containing adaptor inducing interferon IFN-beta (TRIF)-dependent responses. PMID- 18389480 TI - IL-12, IL-23, and IL-27 enhance human beta-defensin-2 production in human keratinocytes. AB - IL-12, IL-23, and IL-27, which are produced by APC, modulate innate and adaptive immunities. Human beta-defensin-2 (hBD-2) produced by epidermal keratinocytes promotes cutaneous antimicrobial defense and inflammation. We examined the in vitro effects of IL-12, IL-23, and IL-27 on hBD-2 production in human keratinocytes. IL-12, IL-23, and IL-27 enhanced IL-1beta-induced hBD-2 secretion and mRNA expression in keratinocytes. The stimulatory effects of IL-12, IL-23, and IL-27 were suppressed by antisense oligonucleotides against NF-kappaB p50 and p65. In addition, the effects of IL-12 and IL-27 were suppressed by antisense STAT3 and STAT1, respectively. All the three IL enhanced the basal and IL-1beta induced transcriptional activities of NF-kappaB, while IL-12 and IL-27 enhanced STAT3 and STAT1 activities, respectively. Further, IL-12, IL-23, and IL-27 promoted basal and IL-1beta-induced phosphorylation of IkappaBalpha. IL-12 and IL 23 tyrosine phosphorylated STAT3 and STAT1, respectively; IL-12, IL-23, and IL-27 tyrosine phosphorylated JAK2 and tyrosine kinase-2; and IL-27 tyrosine phosphorylated JAK1. These results suggest that IL-12, IL-23, and IL-27 may enhance IL-1beta-induced hBD-2 production in keratinocytes by activating NF kappaB. STAT3 and STAT1 are involved in the effects of IL-12 and IL-27, respectively. Thus, IL-12, IL-23, and IL-27 may promote cutaneous antimicrobial defense and inflammation via hBD-2. PMID- 18389481 TI - CD28 ligation in the absence of TCR promotes RelA/NF-kappaB recruitment and trans activation of the HIV-1 LTR. AB - CD28 is one of the most important co-stimulatory receptors necessary for full T lymphocyte activation. CD28 can act as a TCR-independent signalling unit by delivering specific signals which may induce HIV transcription and replication. However, the mechanisms by which CD28 regulates HIV expression remain largely unknown. Here we show that the TCR-independent CD28 signals lead to the trans activation of HIV-1 LTR in an NF-kappaB-dependent manner. In particular, we found that CD28 engagement by B7 induces the specific recruitment of RelA/NF-kappaB subunit to the HIV-1 LTR promoter both in vitro and in ex vivo infected cells. The results obtained by mutating specific tyrosine residues within the CD28 cytoplasmic tail as well as by using LY294002 inhibitory drug evidenced that the recruitment and activation of the phosphatidylinositol 3-kinase/Akt signalling pathway is crucial in mediating CD28-induced HIV transcription through RelA/NF kappaB. PMID- 18389483 TI - Antioxidant and cytoprotective compounds from Berberis vulgaris (barberry). AB - Activity-guided fractionation of an EtOAc-soluble partition of the MeOH extract from the root bark of Berberis vulgaris L. (barberry), using a hydroxyl radical scavenging assay, led to the isolation and identification of three phenolic compounds of a previously known structure, N-(p-trans-coumaroyl)tyramine, cannabisin G and (+/-)-lyoniresinol. Of these, cannabisin G and (+/-) lyoniresinol exhibited antioxidant activity in this bioassay. Furthermore, it was found that cannabisin G showed cytoprotective activity in cultured MCF-7 cells modulated by hydrogen peroxide. PMID- 18389484 TI - Cardioprotective effects and underlying mechanisms of oxymatrine against Ischemic myocardial injuries of rats. AB - Oxymatrine has been demonstrated to have a variety of pharmacological actions. Accumulating evidence indicates that oxymatrine may exert a protective effect on the cardiovascular system. The study was designed to explore the possible role of oxymatrine against myocardial ischemic damage and several related signaling pathways as potential mechanisms. The protective properties of oxymatrine were studied in a rat model of acute myocardial infarction due to permanent ligation of the left anterior descending coronary artery. The results showed that administration of oxymatrine relieved myocardial injuries during ischemia, and this was achieved by protecting cardiomyocytes from apoptotic death. The beneficial effects of oxymatrine were likely mediated by an inhibition of lipid peroxidation (MDA production) and an increase in endogenous antioxidant activity (SOD), activation of the survival signaling molecule (Bcl-2), and a reduction of apoptotic mediator (Fas) and intracellular Ca2+ overload. PMID- 18389485 TI - Tanshinone IIB, a primary active constituent from Salvia miltiorrhiza, exerts neuroprotective effect via inhibition of neuronal apoptosis in vitro. AB - Tanshinone IIB (TSB) is a major active constituent of the roots of Salvia miltiorrhiza (Danshen) widely used in the treatment of stroke and coronary heart disease in Asian countries. This study investigated the in vitro neuroprotective effects of TSB and the underlying mechanism. Co-treatment with TSB significantly inhibited the cytotoxicity and apoptosis of rat cortical neurons induced by staurosporine in a concentration-dependent manner. Consistently, TSB significantly reduced the DNA laddering caused by staurosporine in a concentration-dependent manner. TSB also suppressed the elevated Bax protein and decreased bcl-2 and caspase-3 proteins induced by staurosporine in rat cortical neurons. These findings indicated that TSB had a neuroprotective effect via inhibition of apoptosis. Further studies are warranted to investigate the role of other apoptosis-related signaling proteins and reperfusion-related mechanisms in the protective effect of TSB on neurons. PMID- 18389486 TI - Protective effect of Tinospora cordifolia, Phyllanthus emblica and their combination against antitubercular drugs induced hepatic damage: an experimental study. AB - This study investigated the hepatoprotective effect of two Indian medicinal plants Tinospora cordifolia (Tc), Phyllanthus emblica (Pe), and their combination, in a rat model of isoniazid, rifampicin and pyrazinamide induced hepatic damage. Hepatic damage was assessed using a composite score assigned to histopathological findings of degeneration, necrosis and fibrosis. The antituberculosis treatment (ATT), when given for 90 days, induced significant degeneration and necrosis (score: 7.5; p < 0.01 vs vehicle) associated with morphological changes. However, no change was found in the serum bilirubin and liver enzymes. Co-administration of silymarin (positive control, 50 mg/kg) with ATT protected against necrosis (score: 1.5; p < 0.001 vs ATT). Tc (100 mg/kg) showed a reduction in liver damage (score: 6.5), which was not statistically significant. On the other hand, Pe (300 mg/kg) prevented the necrotic changes to a significant extent (grade 1.0; p < 0.05; score [corrected] 5.5). Combination of Tc and Pe in their therapeutic doses (1:3) significantly prevented the necrosis (score: 3.5; p < 0.001 vs ATT). Similar effects were seen even when the doses were halved and were comparable to the silymarin group. Thus, this study proves the synergistic protective effects exerted by the combination of Tc and Pe when co-administered with ATT. PMID- 18389487 TI - Secretion of mouse growth hormone by transduced primary human keratinocytes: prospects for an animal model of cutaneous gene therapy. AB - BACKGROUND: Keratinocytes are a very attractive vehicle for ex vivo gene transfer and systemic delivery because proteins secreted by these cells may reach the circulation via a mechanism that mimics the natural process. METHODS: An efficient retroviral vector (LXSN) encoding the mouse growth hormone gene (mGH) was used to transduce primary human keratinocytes. Organotypic raft cultures were prepared with these genetically modified keratinocytes and were grafted onto immunodeficient dwarf mice (lit/scid). RESULTS: Transduced keratinocytes presented a high and stable in vitro secretion level of up to 11 microg mGH/10(6)cells/day. Conventional epidermal sheets made with these genetically modified keratinocytes, however, showed a drop in secretion rates of > 80% due to detachment of the epithelium from its substratum. Substitution of conventional grafting methodologies with organotypic raft cultures completely overcame this problem. The stable long-term grafting of such cultures onto lit/scid mice could be followed for more than 4 months, and a significant weight increase over the control group was observed in the first 40 days. Circulating mGH levels revealed a peak of 21 ng/ml just 1 h after grafting but, unfortunately, these levels rapidly fell to baseline values. CONCLUSIONS: mGH-secreting primary human keratinocytes presented the highest in vitro expression and peak circulatory levels reported to date for a form of GH with this type of cells. Together with previous data showing that excised implants can recover a remarkable fraction of their original in vitro mGH secretion efficiency in culture, the factors that might still hamper the success of this promising model of cutaneous gene therapy are discussed. PMID- 18389488 TI - Individualized growth assessment of fetal thigh circumference using three dimensional ultrasonography. AB - OBJECTIVES: To develop individualized growth assessment (IGA) standards for upper (ThC(u)) and middle (ThC(m)) fetal thigh circumferences using three-dimensional ultrasonography. METHODS: A prospective, longitudinal sonographic study of 30 fetuses was performed beginning at 18 weeks' menstrual age. Second-trimester sonographic parameters were measured from three-dimensional volume data to establish IGA standards. Normal infant growth outcomes were confirmed using modified Neonatal Growth Assessment Scores (m(3)NGAS(51)). ThC(u) and ThC(m) were studied in more detail. Rossavik growth model specification procedures, based on the slopes of the second-trimester growth curves, were developed for both ThC(u) and ThC(m). Third-trimester growth trajectories and birth measurements were subsequently predicted for these parameters. Percentage deviations during the third trimester and percentage differences at actual birth age were used to compare observed and predicted measurements. The 95% ranges for Growth Potential Realization Index (GPRI) values for both types of thigh circumference were determined. Values for m(3)NGAS(51) using GPRI(ThC(u)), GPRI(ThC(m)) and GPRI(ThC(o)) (original method) were compared. RESULTS: The 30 newborns had no postnatal evidence of abnormal growth. Two examiners demonstrated a satisfactory measurement bias of mean +/- SD 2.1 +/- 3.6 (95% limits of agreement,-4.9 to 9.1)% for ThC(m) and 3.3 +/- 4.1 (95% limits of agreement,-4.8 to 11.4)% for ThC(u). Rossavik functions fitted parameter trajectories well, with mean R(2) values of 99.5 +/- 0.4% for ThC(u) and 99.6 +/- 0.3% for ThC(m). By fixing coefficients k at their mean values, their respective fits did not change, and the variabilities of coefficients c and s were significantly reduced. For ThC(u), coefficient c was significantly related to the second-trimester slope (R(2)=98.6%), as was s to c(R(2)=91.0%). For ThC(m), coefficient c was significantly related to the second-trimester slope (R(2)=98.6%), as was s to c(R(2)=85.6%). Third-trimester growth trajectories, derived from second-trimester slopes for individual fetuses, had third-trimester deviations of 0.07 +/- 3.7% for ThC(u) and-0.04 +/- 3.7% for ThC(m). Percentage differences at birth age were 16.8 +/- 10.2% for ThC(u) and 8.9 +/- 9.5% for ThC(m). With correction for systematic overestimations, the mean GPRI values were 103.7 (95% range, 90-121)% for ThC(u) and 101.6 (95% range, 88-118)% for ThC(m). Corresponding mean +/- SD m(3)NGAS(51) values, using GPRI(ThC(u)), GPRI(ThC(m)) and GPRI(ThC(o)), were 203 +/- 11%, 201 +/- 10% and 200 +/- 9%, respectively. CONCLUSIONS: Fetal thigh circumference can be measured reliably and evaluated using standard IGA methods. Both ThC(u) and ThC(m) give similar results in the third trimester but neonatal thigh circumference predictions are improved by using ThC(m). Corresponding GPRI(ThC(m)) values are closer to the ideal value of 100% and can be used in m(3)NGAS(51) calculations for assessment of neonatal growth outcome. PMID- 18389489 TI - Antibothropic action of Casearia sylvestris Sw. (Flacourtiaceae) extracts. AB - Casearia sylvestris Sw., popularly known in Brazil as 'guacatonga', has been used as antitumor, antiseptic, antiulcer, local anaesthetic and healer in folk medicine. Snakebite envenomation by Bothrops jararacussu (Bjssu) constitutes a relevant public health hazard capable of inducing serious local damage in victims. This study examined the pharmacological action of apolar and polar C. sylvestris leaf extracts in reverting the neuromuscular blockade and myonecrosis, which is induced by Bjssu venom and its major toxin bothropstoxin-I on the mouse phrenic nerve-diaphragm preparations. The polar methanol extract (ME) was by far the most efficacious. ME not only prevented myonecrosis and abolished the blockade, but also increased ACh release. Such facilitation in neuromuscular transmission was observed with ME alone, but was accentuated in preparations incubated with ME plus venom or toxin. This established synergy opens an interesting point of investigation because the venom or toxin in contact with ME changes from a blocking to a facilitating effect. It is suggested that rutin, known to have potent antioxidant properties, and one of the components present in the ME, could have a role in the observed effects. Since commercial rutin did not reproduce the ME effects, it is likely that a rutin-containing phytocomplex is neutralizing the bothropic envenoming effects. PMID- 18389490 TI - The correlation between angiogenesis and abnormal expression of SERCA2a, phospholamban and the endothelin pathway in heart failure, and improvement by puerarin. AB - Emerging evidence indicates that angiogenesis may be a potential new target in treating heart failure (HF). It was hypothesized that a lack of angiogenesis would correlate with an abnormal expression of sarco/endoplasmic reticulum ATPase 2a (SERCA2a) and phospholamban (PLB), and the activated endothelin (ET) pathway and oxidative stress in HF. If this is the case, such normal changes could be reversed by puerarin. HF was produced by coronary artery ligation for 4 weeks in rats. The rats were divided into three groups: sham, HF untreated and HF + puerarin (120 mg/kg per day, i.p.). Hemodynamic and echocardiographic changes, angiogenesis, cardiac morphology, serum biochemistry, mRNA and proteins of the angiogenesis pathway, the ET pathway and redox were measured. In the HF rats, hemodynamic and echocardiographic abnormalities, cardiac remodeling and histological changes with features of cardiac failure were associated with a lack of the angiogenesis pathway, accompanied by oxidative stress, an up-regulated ET pathway and abnormal SERCA2a and PLB expressions in HF rats. Puerarin significantly promoted angiogenesis and reversed the above changes. In conclusion, the absence of the angiogenesis pathway correlated with abnormal expression of SERCA2a and PLB and an activated ET-ROS (reactive oxygen species) system in the affected myocardium. Puerarin promoted the angiogenesis pathway, improved myocardial microcirculation and down-regulated the ET system, resulting in a reversal of the abnormalities of expression of SERCA2a and PLB, and the cardiac performance in HF. PMID- 18389491 TI - Protective effects of dietary ginger (Zingiber officinales Rosc.) on lindane induced oxidative stress in rats. AB - The protective effect of dietary feeding of Zingiber officinales Rosc. (ginger) against lindane-induced oxidative stress was investigated in male albino rats. Oxidative stress was monitored by estimating the extent of lipid peroxidation, activities of the oxygen free radical (OFR) scavenging enzymes superoxide dismutase (SOD) and catalase (CAT) and the status of the glutathione redox cycle antioxidants. Lindane administration (30 mg/kg bw orally for 4 weeks) was associated with enhanced lipid peroxidation and compromised antioxidant defenses in rats fed a normal diet. Concomitant dietary feeding of ginger (1%w/w) significantly attenuated lindane-induced lipid peroxidation, accompanied by modulation of OFR scavenging enzymes as well as reduced glutathione (GSH) and the GSH dependent enzymes glutathione peroxidase (Gpx), glutathione reductase (GR) and glutathione-S-transferase (GST) in these rats. These findings suggest that a diet containing naturally occurring compounds is effective in exerting protective effects by modulating oxidative stress. PMID- 18389492 TI - Consequences of dichotomization. AB - Dichotomization is the transformation of a continuous outcome (response) to a binary outcome. This approach, while somewhat common, is harmful from the viewpoint of statistical estimation and hypothesis testing. We show that this leads to loss of information, which can be large. For normally distributed data, this loss in terms of Fisher's information is at least 1-2/pi (or 36%). In other words, 100 continuous observations are statistically equivalent to 158 dichotomized observations. The amount of information lost depends greatly on the prior choice of cut points, with the optimal cut point depending upon the unknown parameters. The loss of information leads to loss of power or conversely a sample size increase to maintain power. Only in certain cases, for instance, in estimating a value of the cumulative distribution function and when the assumed model is very different from the true model, can the use of dichotomized outcomes be considered a reasonable approach. PMID- 18389493 TI - Empirically predicted 13C NMR chemical shifts for 8-hydroxyflavone starting from 7,8,4'-trihydroxyflavone and from 7,8-dihydroxyflavone. AB - 8-Hydroxyflavone is not found in nature. While the (13)C chemical shifts of 8 hydroxyflavone have been reported previously, the observed (13)C chemical shifts were not assigned. A previously reported empirical predictive tool has been applied in reverse in order to deconvolute the (13)C chemical shifts for 8 hydroxyflavone from each of those of 7,8,4'-trihydroxyflavone and 7,8 dihydroxyflavone together with those of 7-hydroxyflavone, 4'-hydroxyflavone, and flavone. The two sets of calculated (13)C chemical shifts for 8-hydroxyflavone are in good agreement with each other in that the average absolute difference is 0.4 ppm. The previously reported but unassigned (13)C chemical shifts for 8 hydroxyflavone have been assigned by matching them with the averages of the two sets of calculated (13)C chemical shifts for 8-hydroxyflavone such that the minimum average absolute difference is 0.63 ppm. The assigned (13)C chemical shifts of 8-hydroxyflavone may be used, along with the (13)C chemical shifts of the remaining monohydroxyflavones, as part of a predictive tool to rapidly assess the (13)C NMR spectra of C8-hydroxylated flavonoids. PMID- 18389494 TI - Synthesis, complete 1H and 13C NMR assignment and crystal structure of novel epoxide derivatives of cytochalasin B. AB - Five novel epoxide derivatives of cytochalasin B were synthesized. Reaction of cytochalasin B with t-BHP and BuLi led to selective epoxidation of the C-21/22 double bond to give a single monoepoxide, while reaction with m-CPBA yielded two diepoxides. Reaction of the monoepoxide with m-CPBA yielded two triepoxides. The relative configurations of the epoxides were elucidated by analogy with the natural product by means of spectroscopic methods; full assignment of NMR signals was achieved, and the absolute configuration was confirmed by X-ray crystallography. PMID- 18389495 TI - Frequency-swept HMQC sequences for high-throughput NMR analysis. AB - We describe here new versions of the DEPT phase-encoded HMQC experiment that offer robust performance and improved sensitivity. The new sequences rely on frequency-swept proton and carbon pulses to minimize signal losses from miscalibrated pulses while providing 'J compensation' to optimize the signal strength over a range of heteronuclear coupling constants. By including both proton and carbon-swept pulses, the new sequences also offer an additional signal gain of roughly 10% over well-calibrated hard-pulse experiments. The new sequences also demonstrate that one can construct a sequence that incorporates both 90 degrees and 180 degrees frequency-swept pulses. Although individual pulses in the sequence cause severe phase roll, the phase roll can be eliminated by the proper choice of pulse lengths and sweep directions. PMID- 18389496 TI - Frequency-swept HSQC sequences for high-throughput NMR analysis. AB - This article describes new versions of the DEPT phase-edited heteronuclear single quantum correlation (HSQC) pulse sequence with sensitivity enhancement. The sequences incorporate frequency-swept carbon and proton pulses. The new experiments are inherently robust, well-suited for a high-throughput setting in which sample-to-sample variations may be ignored. The observed signal has the obvious benefit of sensitivity enhancement resulting from the preservation of two magnetization transfer pathways. The two pathways are maintained even in the version of the sequence in which all pulses are frequency-swept. There is an additional signal gain of roughly 10% that derives from the use of both proton and carbon frequency-swept pulses. Furthermore, the sequences use J compensation to provide optimal signal over a range of heteronuclear coupling constants. We demonstrate that the new sequences offer good sensitivity and perform well even when the NMR probe is deliberately mistuned. PMID- 18389497 TI - Rapamycin prevents epilepsy in a mouse model of tuberous sclerosis complex. AB - OBJECTIVE: Tuberous sclerosis complex (TSC) represents one of the most common genetic causes of epilepsy. TSC gene inactivation leads to hyperactivation of the mammalian target of rapamycin signaling pathway, raising the intriguing possibility that mammalian target of rapamycin inhibitors might be effective in preventing or treating epilepsy in patients with TSC. Mice with conditional inactivation of the Tsc1 gene primarily in glia (Tsc1(GFAP)CKO mice) develop glial proliferation, progressive epilepsy, and premature death. Here, we tested whether rapamycin could prevent or reverse epilepsy, as well as other cellular and molecular brain abnormalities in Tsc1(GFAP)CKO mice. METHODS: Tsc1(GFAP)CKO mice and littermate control animals were treated with rapamycin or vehicle starting at postnatal day 14 (early treatment) or 6 weeks of age (late treatment), corresponding to times before and after onset of neurological abnormalities in Tsc1(GFAP)CKO mice. Mice were monitored for seizures by serial video-electroencephalogram and for long-term survival. Brains were examined histologically for astrogliosis and neuronal organization. Expression of phospho S6 and other molecular markers correlating with epileptogenesis was measured by Western blotting. RESULTS: Early treatment with rapamycin prevented the development of epilepsy and premature death observed in vehicle-treated Tsc1(GFAP)CKO mice. Late treatment with rapamycin suppressed seizures and prolonged survival in Tsc1(GFAP)CKO mice that had already developed epilepsy. Correspondingly, rapamycin inhibited the abnormal activation of the mammalian target of rapamycin pathway, astrogliosis, and neuronal disorganization, and increased brain size in Tsc1(GFAP)CKO mice. INTERPRETATION: Rapamycin has strong efficacy for preventing seizures and prolonging survival in Tsc1(GFAP)CKO mice. PMID- 18389498 TI - Development and psychometric evaluation of an Arab version of the Family Peer Relationship Questionnaire. AB - Little is known about parent and peer relationships in immigrant adolescents due to a dearth of culturally sensitive measures. We adapted the Family Peer Relationship Questionnaire (FPRQ) for Arab immigrant mothers and their adolescents. Mothers and adolescents completed parallel versions of the Arab FPRQ and measures of child behavior problems, child social relations, and maternal depression. Data analyses included confirmatory factor analysis and concurrent validity. The proposed four-factor structure was replicated in both the mother and child forms. Scores from the Arab FPRQ subscales and the other measures provided evidence of concurrent and discriminant validity, suggesting that the Arab FPRQ is a viable assessment tool. PMID- 18389499 TI - Randomized clinical trial comparing day-care open haemorrhoidectomy under local versus general anaesthesia. AB - BACKGROUND: Day-care open haemorrhoidectomy under local anaesthesia (LH) may be the most cost-effective approach to haemorrhoidectomy. This prospective randomized trial compared outcome after LH from patients' and clinical perspectives with that after day-care open haemorrhoidectomy under general anaesthesia (GH). METHODS: Forty-one patients with third-degree haemorrhoids were randomized to LH (19) or GH (22). Patient demographics were comparable. A single haemorrhoid was excised in 15 patients, and two and three haemorrhoids in 13 each. Independent nurse-led assessment and clinical evaluation were carried out for 6 months. Outcome measures were mean and expected pain scores at 30, 60 and 90 min, then daily for 10 days, and satisfaction scores at 10 days, 6 weeks and 6 months. Secondary outcomes were journey time within the day-surgery unit and overall cost. RESULTS: Pain was worse following LH than GH at 90 min after surgery (P = 0.028), but pain scores on reaching home were similar. Maximum pain was experienced on day 3 after LH and on day 6 after GH. From day 1 onwards, daily pain scores were lower in the LH group, and there was a significant difference on day 8 (mean (95 per cent confidence interval) 3.61 (2.74 to 4.48) for LH versus 5.29 (4.12 to 6.45) for GH; P = 0.027). Mean pain over 10 days, expectation and satisfaction scores were similar in the two groups. LH had a shorter journey time and was less expensive than GH. CONCLUSION: LH has similar tolerance and clinical outcome to GH, and is associated with a shorter journey time and lower cost. REGISTRATION NUMBER: NCT00503269 (http://www.clinicaltrials.gov). PMID- 18389500 TI - Hearing in a diurnal, mute butterfly, Morpho peleides (Papilionoidea, Nymphalidae). AB - Butterflies use visual and chemical cues when interacting with their environment, but the role of hearing is poorly understood in these insects. Nymphalidae (brush footed) butterflies occur worldwide in almost all habitats and continents, and comprise more than 6,000 species. In many species a unique forewing structure- Vogel's organ--is thought to function as an ear. At present, however, there is little experimental evidence to support this hypothesis. We studied the functional organization of Vogel's organ in the common blue morpho butterfly, Morpho peleides, which represents the majority of Nymphalidae in that it is diurnal and does not produce sounds. Our results confirm that Vogel's organ possesses the morphological and physiological characteristics of a typical insect tympanal ear. The tympanum has an oval-shaped outer membrane and a convex inner membrane. Associated with the inner surface of the tympanum are three chordotonal organs, each containing 10-20 scolopidia. Extracellular recordings from the auditory nerve show that Vogel's organ is most sensitive to sounds between 2-4 kHz at median thresholds of 58 dB SPL. Most butterfly species that possess Vogel's organ are diurnal, and mute, so bat detection and conspecific communication can be ruled out as roles for hearing. We hypothesize that Vogel's organs in butterflies such as M. peleides have evolved to detect flight sounds of predatory birds. The evolution and taxonomic distribution of butterfly hearing organs are discussed. PMID- 18389501 TI - Pharmacogenetics of parkinsonism, rigidity, rest tremor, and bradykinesia in African-Caribbean inpatients: differences in association with dopamine and serotonin receptors. AB - We studied the association between polymorphisms of genes coding for dopamine D(2) (DRD2), dopamine D(3) (DRD3), serotonin 2(a) (HTR2A), and serotonin 2(c) (HTR2C) receptors and Antipsychotic-Induced Parkinsonism (AIP), rigidity, bradykinesia, and rest-tremor in African-Caribbeans treated with antipsychotics. Polymorphisms of DRD2 (-141CIns/Del, TaqIA, 957C > T), DRD3 (Ser9Gly), HTR2A ( 1438A > G, 102T > C, His452Tyr), and HTR2C (-759C > T, Cys23Ser) genes were determined according to standard protocols. The Unified Parkinson Disease Rating Scale was used for the measurement of AIP, rigidity, bradykinesia, and rest tremor. Chi-squared or Fisher's exact tests were applied for the association analyses. The t-test was applied for continuous data. Ninety nine males and 27 females met the inclusion criteria (Schizophr Res 1996, 19:195). In males, but not in females, there were significant associations between -141CDel-allele carriership (DRD2) and rigidity (Fisher's Exact Test: P = 0.021) and between 23Ser-allele carriership (HTR2C) and bradykinesia (P = 0.026, chi(2) = 5.0) or AIP (P = 0.008, chi(2) = 7.1). Rest-tremor was not associated with any of the polymorphisms studied. Analyses of the age, chlorpromazine equivalents, benztropine equivalents, the number of patients using anticholinergic medication, and the utilization patterns of the antipsychotic medication did not show statistically significant differences between patients with and without AIP, rigidity, bradykinesia, rest-tremor. Conducting the analysis without gender stratification did not affect our findings considerably, except for the association between bradykinesia and 23Ser-allele which failed to reach statistical significance in the total sample (P = 0.0646, chi(2) = 3.41). Since AIPs subsymptoms (rigidity, bradykinesia, and rest-tremor) may differ pharmacogenetically, our data strongly support symptom-specific analysis of AIP. However, further research is warranted to confirm our findings. PMID- 18389503 TI - Effect of surface decoration of CNTs on the interfacial interaction and microstructure of epoxy/MWNT nanocomposites. AB - Epoxy resin nanocomposites with different contents of multiwalled carbon nanotubes (MWNTs) are prepared. The interaction between MWNTs and the epoxy resin matrix and the microstructure and mechanical properties of the composites are systematically investigated by Fourier-transform infrared (FTIR) spectroscopy, transmission electron microscopy, scanning electron microscopy, and positron annihilation technology. FTIR spectra reveal that two kinds of hydrogen bonds exist at the interface for the nanocomposites modified by amine, one between the epoxy group on the side chain and the NH group, and the other between the epoxy group on the alicycle and the NH group. Compared to unmodified MWNT composites, the modified MWNT composites possess better mechanical properties, which are attributed to stronger interfacial interaction resulting from an efficient load transfer from matrix to MWNTs. Positron annihilation lifetime spectroscopy is used to characterize the microstructure of the epoxy/MWNT composites. The subtransition and glass transition temperatures are determined by finite-term positron lifetime analysis and the variation of the free-volume size as a function of temperature. Shifts of structure transition temperatures of the composites are observed with increasing MWNT weight content. Interestingly, the continuous lifetime analysis reveals the existence of two long-lived lifetime components above the glass transition temperature, which may be attributed to the formation of local ordered regions related to the packing density of chains. PMID- 18389502 TI - Mammary gland differentiation inversely correlates with GDF-8 expression. AB - GDF-8 is recognised as an inhibitor of muscle cell growth and differentiation. Although initially thought to be restricted to muscle cells it is now accepted that GDF-8 expression has a broader tissue distribution. We demonstrate GDF-8 expression in the mouse mammary gland, which is predominantly associated with epithelial cells and displays an inverse correlation to the differentiated state of the gland. Specifically, the highest GDF-8 mRNA levels correlate with periods of maximal ductal growth, diminish as pregnancy progressed and are down-regulated to minimal levels by the onset of lactation as the epithelium differentiates. A similar profile is observed for both GDF-8 protein processing and reflects Smad2/3 phosphorylation profile. However, in contrast to muscle cells, GDF-8 neither reduces proliferation nor induces p21 expression levels in mammary epithelial cells. These data implicate a role for GDF-8 in mammary epithelial cell differentiation and demonstrate that GDF-8 has cell-type specific activities. PMID- 18389504 TI - An iminoboronate construction set for subcomponent self-assembly. AB - Recently we have demonstrated a series of systems in which complex structures were created from simple amine and aldehyde subcomponents by copper(I)-templated imine bond formation. We describe herein the extension of this "subcomponent self assembly" concept to the generation of structures based upon the iminoboronate ester motif. Equimolar amounts of diol, amine, and 2-formylphenylboronic acid reacted by reversible B-O and C=N bond formation to generate iminoboronate esters, as has recently been reported by James et al. (Org. Lett. 2006, 8, 609 612). The extent of ester formation was shown to depend upon a number of factors. The exploration of these factors allowed rules and predictions to be formulated governing the self-assembly process. These rules allowed the construction of more complex structures containing multiple boron atoms, including a trigonal cage containing six boron centers, as well as pointing the way to the construction of yet more intricate architectures. The lability of the B-O and C=N bonds also allowed different diol and amine subcomponents to be substituted within these structures. Selection rules were also determined for these substitution reactions, allowing the products to be predicted based upon the electronic properties of the diols and diamines employed. These results thus demonstrate the generality of the subcomponent self-assembly methodology through its application to a new dynamic covalent system. PMID- 18389505 TI - Asymmetric total synthesis of PDIM A: a virulence factor of Mycobacterium tuberculosis. PMID- 18389506 TI - Variably isotactic poly(hydroxybutyrate) from racemic beta-butyrolactone: microstructure control by achiral chromium(III) salophen complexes. PMID- 18389507 TI - Yeast cells with an artificial mineral shell: protection and modification of living cells by biomimetic mineralization. PMID- 18389508 TI - Dynamics in the p38alpha MAP kinase-SB203580 complex observed by liquid-state NMR spectroscopy. PMID- 18389510 TI - Catalytic asymmetric C-Si bond formation to acyclic alpha,beta-unsaturated acceptors by Rh(I)-catalyzed conjugate silyl transfer using a Si-B linkage. PMID- 18389511 TI - Protein surface recognition: structural characterisation of cytochrome c porphyrin complexes. PMID- 18389509 TI - Isolation, structure elucidation, and biomimetic total synthesis of versicolamide B, and the isolation of antipodal (-)-stephacidin A and (+)-notoamide B from Aspergillus versicolor NRRL 35600. PMID- 18389512 TI - Theoretical study on the complexes of benzene with isoelectronic nitrogen containing heterocycles. AB - The pi-pi interactions between benzene and the aromatic nitrogen heterocycles pyridine, pyrimidine, 1,3,5-triazine, 1,2,3-triazine, 1,2,4,5-tetrazine, and 1,2,3,4,5-pentazine are systematically investigated. The T-shaped structures of all complexes studied exhibit a contraction of the C--H bond accompanied by a rather large blue shift (40-52 cm(-1)) of its stretching frequency, and they are almost isoenergetic with the corresponding displaced-parallel structures at reliable levels of theory. With increasing number of nitrogen atoms in the heterocycle, the geometries, frequencies, energies, percentage of s character at C, and the electron density in the C--H sigma antibonding orbital of the complexes all increase or decrease systematically. Decomposition analysis of the total binding energy showed that for all the complexes, the dispersion energy is the dominant attractive contribution, and a rather large attraction originating from electrostatic contribution is compensated by its exchange counterpart. PMID- 18389513 TI - Intrachain versus interchain electron transport in poly(fluorene-alt benzothiadiazole): a quantum-chemical insight. AB - Poly(9,9-di-n-octylfluorene-alt-benzothiadiazole) [F8BT], displays very different charge-transport properties for holes versus electrons when comparing annealed and pristine thin films and transport parallel (intrachain) and perpendicular (interchain) to the polymer axes. The present theoretical contribution focuses on the electron-transport properties of F8BT chains and compares the efficiency of intrachain versus interchain transport in the hopping regime. The theoretical results rationalize significantly lowered electron mobility in annealed F8BT thin films and the smaller mobility anisotropy (mu( parallel)/mu( perpendicular)) measured for electrons in aligned films (i.e. 5-7 compared to 10-15 for holes). PMID- 18389514 TI - Dipeptide derivatives of AZT: synthesis, chemical stability, activation in human plasma, hPEPT1 affinity, and antiviral activity. AB - 5'-O-Dipeptide ester prodrugs of antiviral zidovudine (AZT) were designed to target the human intestinal oligopeptide transporter, hPEPT1, and were evaluated for their stability at pH 7.4 in buffer and in human plasma, affinity toward hPEPT1, cytotoxicity, and antiretroviral activity. The dipeptide esters of AZT undergo cyclization in buffer at pH 7.4 to release the parent drug at a rate that depends on the size of the side chains of the peptide carrier; the prodrug is considerably more stable if bulky beta-branched amino acids such as Ile and Val are present, particularly as C-terminal residues. Incubation in human plasma showed that most of the dipeptide esters of AZT release the parent drug through two aminopeptidase-mediated pathways: 1) stepwise cleavage of each of the amino acids and 2) direct cleavage of the dipeptide-drug ester bond. However, the plasma hydrolysis of Gly-Gly-AZT and Phe-Gly-AZT showed only direct cleavage of the dipeptide-drug ester bond. Substrate half-lives in plasma were again remarkably high when hydrophobic beta-branched amino acids (Val, Ile) were present. The esters were also good substrates for the intestinal oligopeptide transporter hPEPT1 in vitro, with Val-Gly-AZT and Val-Ala-AZT presenting the highest affinity toward the transporter (IC(50): 0.20 and 0.15 mM, respectively). The AZT dipeptide esters were assayed against the IIIB and ROD strains of HIV, and their cytotoxicity was evaluated in MT-4 cells. The selectivity index of the prodrugs was two- to threefold higher than that of AZT for all compounds analyzed. These results point to the potential of dipeptide-based carriers for the development of effective antiviral drug-delivery systems. Val-Ala-AZT appears to combine chemical stability with good affinity for the hPEPT1 transporter and an improved cytotoxicity/antiretroviral index relative to AZT. PMID- 18389515 TI - Synthesis and anti-inflammatory activity evaluation of novel 7-alkoxy-1-amino-4,5 dihydro[1,2,4]triazole[4,3-a]quinolines. AB - In this study, a novel series of 7-alkoxy-1-amino-4,5-dihydro[1,2,4]triazole[4,3 a]quinolines were synthesized by using 6-hydroxy-3,4-dihydro-2(1H)-quinolone as the starting material. These compounds were evaluated for anti-inflammatory activity through monitoring their ability to inhibit xylene-induced ear edema in mice. Some of the tested compounds exhibited significant activity, and the compounds 5f (7-(benzyloxy)-4,5-dihydro[1,2,4]triazolo[4,3-a]quinolin-1-amine) and 5i (7-(p-chlorobenzyloxy)-4,5-dihydro[1,2,4]triazolo[4,3-a]quinolin-1-amine) showed the highest anti-inflammatory activity (52% and 58% inhibition, respectively, at 2 h pre-administration) which were comparable to or even slightly more potent than the reference drug ibuprofen (55%). Furthermore, the structure-activity relationship of these 1,2,4-triazole quinolines was demonstrated. PMID- 18389516 TI - Thalidomide as a multi-template for development of biologically active compounds. AB - Thalidomide is a teratogenic/hypnotic/sedative agent which elicits a wide range of pharmaceutical/biological activities. The diversity of its biological activities suggested that the drug might be useful as a multi-template for development of various kinds of biologically active compounds. We adopted two strategies for the structural development of thalidomide. The first was to develop the structure of the drug based on the target molecules to which thalidomide itself and/or its metabolites directly bind, or the assay systems in which thalidomide itself and/or its metabolites exhibit activity. Based on this strategy, tumor necrosis factor-alpha production-regulating agents, cyclooxygenase inhibitors, nitric oxide synthase inhibitors, histone deacetylase inhibitors, anti-angiogenic agents, and tubulin polymerization inhibitors have been created. The second was to develop the structure of thalidomide based on hypothetical target molecule(s)/biological response(s) which might be relevant to the pharmacological effects elicited by thalidomide. Based on this strategy, androgen antagonists, progesterone antagonists, cell differentiation inducers, aminopeptidase inhibitors, thymidine phosphorylase inhibitors, mu-calpain inhibitors, alpha-glucosidase inhibitors and nuclear liver X receptors (LXRs) antagonists have been created. Our structural development studies on thalidomide are reviewed focusing on recent development of tubulin polymerization inhibitors, alpha-glucosidase inhibitors, and nuclear liver X receptors antagonists. PMID- 18389517 TI - QSAR study on dual 5-HT1A and 5-HT1B antagonists: an insight into the structural requirement for antidepressant activity. AB - The 5-HT autoreceptors have received considerable attention as potential targets for the development of antidepressants. With the purpose of designing new chemical entities with enhanced antagonist potencies against 5-HT1A and 5-HT1B, a QSAR study carried out on thienopyrimidinone derivatives as antagonists of serotonin autoreceptors is presented. The developed models were validated by standard QSAR parameters and through a detailed structural analysis on how the QSARs reproduce and explain the differences in the experimentally known activity data. The developed models showed a good correlative and predictive ability having a squared cross validated correlation co-efficients (r2 cv) of 0.780 for 5 HT1A and 0.638 5-HT1B antagonism. The squared conventional correlation co efficients (r2) were found to be 0.824 for the 5-HT1A model and 0.745 for 5-HT1B antagonism. The study indicated that the 5-HT autoreceptor antagonistic activity exhibited by the series is largely explained by steric factors of substituents which underline the role of size and shape of thienopyrimidinones in making effective antagonist-autoreceptor interaction chemistry. A detailed comparative investigation was made between the two models and the insights gleaned from the study could be usefully employed to design dual antagonists with a much more enhanced potency and selectivity. PMID- 18389518 TI - Cytotoxic-antineoplastic derivatives of prenyl-1,2-naphthohydroquinone. AB - Several new prenyl-1,2-naphthohydroquinone derivatives have been prepared by chemical modifications of Diels-Alder products which were obtained from cycloaddition of alpha-myrcene to 1,2-benzoquinone and then evaluated in vitro for their cytotoxic activity against A-549 lung carcinoma, HT-29 colon carcinoma, and MB-231 breast adeno-carcinoma culture cells. Most of them exhibited GI50 values in the microM-concentration level. PMID- 18389519 TI - Supercritical fluid extraction of ecdysterone from the roots of Achyranthes bidentata BL. AB - Ecdysterone has been found in a great many plants and animals and has some valuable pharmaceutical properties. The present study was conducted to investigate optimal conditions for the extraction of the compound by supercritical fluid extraction from the roots of Achyranthes bidentata BL. An orthogonal array design (OAD), OA(9)(3(4)), was employed as a chemometric method for optimization of the extraction of ecdysterone from the herbal medicine. Four parameters, namely, pressure and temperature of the supercritical fluid, the dynamic extraction time, and the flow rate of dimethyl sulfoxide, were studied and optimized by a three-level OAD. Determinations of the extracts were performed by high-performance liquid chromatography. The effects of the parameters were studied using analysis of variance. The results shown that the yield of ecdysterone could be influenced by the four parameters to a similar degree. The yield for DMSO-modified supercritical CO(2) was in the range from 0.65 to 1.03 mg/g under the selected conditions. In comparison with methanol-modified supercritical CO(2 )and Soxhlet extraction, a higher yield was obtained when DMSO modified supercritical CO(2) was used. PMID- 18389520 TI - Effect of pressure drop on solute retention and column efficiency in supercritical fluid chromatography. Part 2: Modified carbon dioxide as mobile phase. AB - The effect of pressure drop on the performance of supercritical fluid chromatography systems using a modified mobile phase (carbon dioxide + ethanol) was studied. Experiments were performed on a Lichrospher-RP-18 column with phenanthrene as a solute. A wide range of back pressures (130 to 210 bar) and modifier concentrations (2 to 7% w/w) have been explored. Experiments yielding both small and large pressure drops were performed. From these experiments, parameters to describe pressure drop, retention, and column efficiency were extracted, and were used to simulate the dynamics of the chromatographic column. A good match between the experimentally measured and calculated values of pressure drop, retention times, and column efficiency was observed. At low back pressure and modifier composition, significant loss of column efficiency was observed. PMID- 18389521 TI - Ethnic differences in prevalence of bipolar disorder in Te Rau Hinengaro: the New Zealand Mental Health Survey. PMID- 18389522 TI - The use of xenograft models for the selection of cancer treatments with the EGFR as an example. AB - Mouse models of cancer have consistently been used to qualify new anti-cancer drugs for development of human clinical trials. The most used models are xenografts of human tumors grown subcutaneously in immunodeficient mice such as athymic (nude) or severe combined immune deficient (SCID) mice. However, the number of anti-cancer agents that fail in the clinic far outweighs those considered effective, suggesting that the selection procedure for progression of molecules into the clinic requires improvement. This has provoked considerable skepticism about the value of using such preclinical models. As a result, a shift has occurred towards developing and using spontaneous mouse tumor arising in transgenic and/or knockout mice engineered to recapitulate various genetic alterations thought to be causative of specific types of human cancers. Alternatively, the option has been to improve human tumor xenograft models by using orthotopic transplantation and, therefore, promotion of metastatic spread of the resultant 'primary' tumors. Here we review the value and the limitations of xenograft models and their role in developing new anti-cancer treatments. PMID- 18389523 TI - Platelet-activating factor, PAF acetylhydrolase, and anaphylaxis. PMID- 18389524 TI - Platelet-activating factor, PAF acetylhydrolase, and anaphylaxis. PMID- 18389525 TI - Patent foramen ovale and cryptogenic stroke. PMID- 18389526 TI - Patent foramen ovale and cryptogenic stroke. PMID- 18389527 TI - Patent foramen ovale and cryptogenic stroke. PMID- 18389528 TI - Adolescent overweight and coronary heart disease. PMID- 18389529 TI - Mayston comment. PMID- 18389530 TI - Problems in management of cerebral palsy in a developing country. PMID- 18389531 TI - Cochlear implants. PMID- 18389532 TI - The definition of cerebral palsy, April 2006. PMID- 18389533 TI - Biotech in Bangalore. PMID- 18389537 TI - More education is recommended before teaching. PMID- 18389543 TI - Influenza vaccine a matter of choice? PMID- 18389545 TI - Global warming questioned. PMID- 18389544 TI - Memorial project strengthens nurse's commitment. PMID- 18389546 TI - Effectiveness of physical therapy for low back pain. PMID- 18389548 TI - [Anorexia nervosa in men, an ignored reality]. PMID- 18389549 TI - [Refeeding syndrome in the treatment of anorexia]. PMID- 18389550 TI - [Visual plasticity in children]. PMID- 18389551 TI - [Vision disorders in children: what type of genetic counseling?]. PMID- 18389552 TI - [Optical equipment for children. Point of view of the optician]. PMID- 18389553 TI - [Rehabilitation of strabismus in children]. PMID- 18389554 TI - [Contact lenses in children]. PMID- 18389555 TI - [Refractive surgery in children]. PMID- 18389556 TI - Somalia is again polio-free. PMID- 18389557 TI - Validation of neonatal tetanus elimination in Zambia by lot quality-assurance cluster sampling. PMID- 18389558 TI - Modern sport, modernism and the cultural manifesto: de Coubertin's Revue Olympique. AB - This article studies the Modern Olympic Movement within the cultural framework of early twentieth-century aesthetics. By examining the first 13 years of the Revue Olympique, the bond between sport and ideals of beauty emerges as a powerful indicator of a broader socio-cultural discourse. Discourse analysis exposes the Revue Olympique as homogeneous and coherent device of praxis that located the Olympic Movement and the genre of sport festival it promoted among the dominant ideas about beauty, art and culture of this era. It functioned as a type of cultural manifesto for Pierre de Coubertin's international Olympic Movement. PMID- 18389559 TI - The agricultural labourer and the "hodge" stereotype, c. 1850-1914. PMID- 18389560 TI - Feminism in London, circa 1850-1914. PMID- 18389562 TI - Household and market in suffragette discourse, 1903-14. PMID- 18389563 TI - Population growth, poverty and foodgrain supply in India: the present trend and future prospect. PMID- 18389564 TI - Effectiveness of nitroblue tetrazolium (NBT) test. PMID- 18389565 TI - Information on IMIA. PMID- 18389566 TI - Asystole during electroconvulsive therapy in an elderly woman treated concomitantly with venlafaxine. PMID- 18389567 TI - Single ion channels in the slime mold Dictyostelium discoideum. AB - Single ion channels with different conductances and gating characteristics were observed in the plasma membrane of the slime mold Dictyostelium discoideum by means of the patch-clamp technique in the cell-attached mode. The predominant channel type shows outward current flow, probably carried by K+ ions. The slope conductance of this channel is 9 pS and its probability to be open increases with depolarization of the membrane. The channel is observed from 1 to 8 h after the beginning of starvation. PMID- 18389569 TI - Psychosocial factors in research on work conditions and health in Sweden. PMID- 18389568 TI - Broad overview of the history of Swedish occupational health research. PMID- 18389570 TI - International evaluation of Swedish work environment research. PMID- 18389571 TI - China's fertility transition through regional space: using GIS and census data for a spatial analysis of historical demography. PMID- 18389572 TI - Lessons from the evaluations of Swedish work environment and public health research. PMID- 18389573 TI - Preventing accidents and injuries in the MRI suite. PMID- 18389574 TI - [Presentation. Introduction]. PMID- 18389575 TI - Vaccination considered the only practical safeguard against bluetongue. PMID- 18389576 TI - Putting research into practice. PMID- 18389577 TI - [Certain biochemical alterations in the blood of patients with atherosclerosis obliterans of the lower limb arteries]. AB - The article reviews evidence regarding certain biochemical alterations in the blood of patients diagnosed with atherosclerosis obliterans of the lower limb arteries (AOLLA) before and after surgical treatment. Also analyzed herein are alterations in the levels of nitrogen oxide (as a marker of the endothelial function) and the activity of superoxide dismutase (as one of the key enzymes of the antioxidant system) in blood serum of patients with AOLLA. Statistically significant alterations in the above-mentioned parameters depending on the type of the operative treatment were revealed. PMID- 18389579 TI - The working-class smoker. PMID- 18389578 TI - [Definition, classification, rationale and treatment for PCl deficient knee]. PMID- 18389580 TI - A sisterhood of suffering. PMID- 18389581 TI - The myth of 'best in the world'. PMID- 18389582 TI - Hospitals as budget target. Bush's proposed budget has hospitals crying foul over a 'savings' plan that reduces Medicare, Medicaid funding by $200 billion. AB - President Bush's fiscal 2009 budget proposal comes equipped with $200 billion in Medicare and Medicaid savings. While the Bush administration deems the figure necessary to ensure the programs remain solvent, hospitals are crying foul. Michael Connelly, left, president and CEO of Catholic Healthcare Partners, said Bush's budget proposal "fundamentally missed the point." PMID- 18389583 TI - Judgment day for NCFE. Defense cites sluggish government reimbursement. PMID- 18389585 TI - Baylor wins NQF quality award. Corrigan: System 'stood out as an exemplary model for raising the bar'. PMID- 18389584 TI - Q: Who will follow the leader? A: Good question. Leadership survey reveals hospitals aren't doing enough to groom future top execs. PMID- 18389586 TI - Lessons from a debacle. Calif. reform defeat, the latest in the states, spells trouble for universal coverage. PMID- 18389587 TI - The industry responds on privacy. Coalition of healthcare companies is working on new protections for patient data. PMID- 18389588 TI - Trustees of the year 2008. 'We are going to do the job for them'. PMID- 18389589 TI - Trustees of the year 2008. Healing a hospital. PMID- 18389590 TI - More doubts over reporting. PMID- 18389591 TI - [Modern concept of gastroesofageal reflux disease: from Genvaal to Monreal]. PMID- 18389592 TI - [Clinical experimental ground of positive influence of hypotensive drugs on defect of the gastric mucosa in stomach ulcer]. PMID- 18389593 TI - [Effect of oral intake of feeding formulas on metabolic activity of the gut microflora in patients subjected to stomach and small intestine resection]. PMID- 18389594 TI - [Effect of ursodeoxycholic acid on the characteristics of lipid metabolism in cholelithiasis and gall-bladder cholesterosis]. AB - The article represents data demonstrating influence of ursodeoxicholic acid on the characteristics of lipidic interchange during cholesterin assosiated diseases of biliary tract. It has been shown that hypercholesterolemias occur in 64% of cases under cholelithiasis and in 56% of cases under gall-bladder cholesterosis. Slow rising of level of total cholesterol (within 5.3-6.1 mmole/l) is registered in 68% of cases under cholelithiasis and in 69% of cases under gall-bladder cholesterosis, higher hypercholesterolemia occurs rare and doesn't exceed 2-4% of cases. Disappearance of hypercholesterolemia after 3-month course of treatment with Ursosan has been registered in 45% of patients with cholelithiasis and in 49% of patients with gall-bladder cholesterosis. Other patients showed positive dynamics, and level of total cholesterol made up in general 5.6 mmoles/l under cholelithiasis and 5.4 mmoles/l uder gall-bladder cholesterosis. PMID- 18389595 TI - [Etiology of chronic diffuse liver damages]. AB - Patients with chronic diffuse liver damages have cases of chronic hepatitis associated with alcoholic liver disease being transformed to cirrhosis 2.1 times more often when having mostly equal frequency of chronic damage of viral and alcoholic origin on the stage of chronic hepatitis or in the case of pure viral damage against a background of significant prevalence (3 times) of viral etiology. Infection associated with HCV in patients with alcoholic liver cirrhosis doesn't significantly influence the course of disease, aftereffects and prognosis of patients. PMID- 18389596 TI - [Chronic HBeAg-negative viral gepatitis B: treatment is essential and possible]. PMID- 18389597 TI - [Effect of zinc deficiency on effectiveness of the treatment of liver encephalopathy in patients with liver cirrhoses]. AB - Zinc plays an important role in human physiologic processes being cofactor of many enzymes, it participates in numerous enzyme reactions. We have investigated 15 patients with liver cirrhoses of different etiology. With the rising of class of liver impairment and developing liver encephalopathy, level of zinc in blood drops. Treatment with diuretic therapy results in increase of discharge of zinc with urine and prevents recovery of its level in blood. PMID- 18389598 TI - [Functional renal reserve of patients with liver cirrhoses]. PMID- 18389599 TI - [Correction of portal hypertension by beta-adrenoblockers (atenolol and metoprolol) and inhibitors of ACE (lisinopril and enalapril) in liver cirrhosis]. PMID- 18389600 TI - [Pharmacological study of herbal preparation for complex treatment of intestine dysbacteriosis]. PMID- 18389601 TI - [Hypothalamic control of the myoelectric activity of the gastric antrum in rabbits during acute emotional stress]. AB - During the experiments on the rabbits immobilized, which were undergone one day food deprivation, the influence of electric irritation of the lateral hypothalamus "hunger center" in condition of the acute emotional stress, that appears under unreinforcement of the artificially amplified hunger motivated excitation with food, which is located on the close but not reachable distance from animals, has been studied. It has been proved that increase of the alimentary motivation intensity in the process of the double-side turn-by-turn electric stimulation of digestive hypothalamic centers and appearance of uneffective food intake behaviour leads to rapid change of the bimodal patterns of the basic electric rhythm of the stomach distribution on the three-modal type distribution, which is typical for the state of hunger in condition of two days' food deprivation and also to the appearance of ectopic focus of excitations in the antral stomach part, and finally--to the development of ulcer-erosive injuries in the stomach mucosa. PMID- 18389602 TI - [Experimental study of NSAID effect on prostaglandine contents]. PMID- 18389603 TI - [Optimization of the treatment of stomach ulcer in patients subjected to perforated gastroduodenal ulcer closure]. AB - The organized benchmark analysis result treatments 152 sick with taken in perforated gastroduodenal ulcers. In the main group (87 sick) in complex treatment are used methods of ozonotherapy. In group of the comparison (65 sick) used standard scheme of the treatment to peptic ulcer. Methods of ozonotherapy promote more early and more full recovering the structure gastroduodenal mucous and provide efficient eradication of Helicobacter pylori, reduces the time of the healing the ulcers and reduces the risk of the relapse ulcers. PMID- 18389604 TI - [The role of translocation of intestine bacterial autoflora and its toxic biomolecules in human pathology]. PMID- 18389605 TI - [New approach to the treatment and prophylaxis of NSAID-induced gastropathies in ambulatory practics]. PMID- 18389606 TI - [Treatment experience of patients with irritable bowel syndrome complicated by constipations with medicinal preparation softovak]. PMID- 18389607 TI - [Contemporary scopes of diagnostics of enterohepatic circulation of bile acids and profilaxys of cholecystolithiasis]. PMID- 18389608 TI - [Clinical course of Helicobacter pylori associated chronic gastritis in post eradicational period]. AB - Results of a prospective cohort study of Helicobacter pylori associated chronic gastritis in posteradicational period have been analyzed in the paper. Different versions and prognostic markers in posteradicational period were identified. Curation algorithm for a patient with chronic gastritis after antihelicobacter therapy was suggested. PMID- 18389609 TI - [The role of essential phospholipids in regulation of functional systems]. PMID- 18389610 TI - [Early prognosis of hormone refractoriness development in inflammatory intestinal diseases]. PMID- 18389611 TI - [Clinical pathogenetic characteristics of chronic gastritis associated with bile excreting system diseases]. PMID- 18389612 TI - [Sergei Petrovich Botkin--founder of physiological school in Russian medicine (to 175th anniversary of birthday)]. PMID- 18389613 TI - [Case study of acute toxic hepatitis associated with mushroom poisoning of infant baby]. PMID- 18389615 TI - [Circulating deoxyribonucleic acids in blood and their using in medical diagnostics]. AB - Circulating nucleic acids were discovered more then 30 years ago and were intensively investigated for the last ten years. The manuscript review the data regarding sources of circulating deoxyribonucleic acids in blood, factors influence on their circulation, like blood nucleases, and excretion of DNA from blood. Application of circulating DNA in practical medicine is also discussed. PMID- 18389614 TI - [Molecular-genetic basis of tapetoretinal degeneration]. AB - The review on problem tapetoretinal degeneration (TD) which represents serious enough and incurable disease revealed with frequency 1 : 3500-5000 in general population is presented. The most often reason of occurrence TD are mutations in RHO, RDS and RPE65 genes. The precise interrelation of pigmentary degenerations of a retina and mutations in genes RHO, RDS and RPE65 will allow to develop approaches of DNA--diagnostics of hereditary dystrophies of a retina so frequently meeting in clinical practice of the ordinary ophthalmologist, and also to pass at medical genetics consultation from probability estimations of risk of disease to unequivocal. Also the molecular analysis of genes changes in the providing correct functioning of photoreceptors and pigmentary epithelium of a retina and determining pathological changes at TD, will allow to approach to understanding of the physiological and pathological processes proceeding in a retina and by that will serve becoming and development pathogenic to caused therapy TD closer. PMID- 18389616 TI - [Nucleotide excision repair in mammalia: mechanism of a primary damage recognition]. AB - Nucleotide excision repair is one of the most important pathways of DNA repair in eukaryotic cells. Defects of this system lead to serious diseases including certain kinds of cancer. Nucleotide excision repair is able to remove a wide range of the structurally diverse DNA damages such as UV induced pyrimidine dimers, bulky chemical adducts arising under the action of carcinogenic compounds or chemotherapeutical drugs on cellular DNA. A broad substrate specificity of this repair pathway is related to the main intriguing question that is the mechanism of damage recognition by the protein complex in the context of the large excess of undamaged DNA. This review is detailed on the key stage of nucleotide excision repair--the recognition of a lesion in DNA, which is still most debated. We have considered the main models of a primary damage recognition and preincision complex formation that have been suggested by the leading groups in this field. Data presented allow to suggest the model of sequential loading of the proteins of reparative complex on damage DNA as the most reasonable. PMID- 18389617 TI - [Association of cytochrome P450 genes polymorphisms (CYP1A1 and CYP1A2) with the development of chronic obstructive pulmonary disease in Bashkortostan]. AB - To assess the role that polymorphisms of cytochrome P450 genes play in genetic predisposition to chronic obstructive pulmonary disease (COPD), the allele and genotype distributions of CYPIA1 (2455 A/G, 3801T/C) and CYP1A2 (-2464T/delT, 163C/A) genes were studied in Tatar and Russian COPD patients and in cases of healthy individuals (Russian, Tatar and Bashkir), residents of Bashkortostan. It was shown that the CYP1A1 and CYP1A2 genes haplotypes frequency distribution patterns do not differed between Tatars and Russians ethnic groups (chi2 = 0.973, df = 3, p = 1.00 and chi2 = 1.546, df = 3, p = 0.92, respectively). Analysis of the the CYP1A1 and CYP1A2 genes haplotypes revealed statistically significant differences in the haplotypes frequency distributions between Bashkirs versus Russians and Tatars (chi2 = 12.328, df= 3,p = 0.008; chi2 = 9.218, df=3, p = 0.034, respectively for CYP1A1 gene and (chi2 = 18.779, df=3, p = 0.0001, chi = 14.326, df=3, p = 0.003, respectively for CYP1A2 gene). The (-2467)delT allele and CYP1A2*1D haplotype of CYPIA2 gene was associated with higher risk of COPD in Tatar ethnic group (OR = 1.83, 95% CI 1.24-2.71, chi2 = 9.48, p = 0.003 and chi2 = 9.733, p = 0.0027, Pcor = 0.008; OR = 3.908, 95% CI 1.56-10.19, respectively). On the other hand the CYP1A2*1A haplotype had protective effect (chi2 = 6.319, p = 0.0127, Pcor = 0.038; OR = 0.6012, 95% CI 0.402-0.898). But at the same time we did not find any differences in the genotypes and haplotypes frequency distributions of the CYP1A2 gene within the patients and healthy groups in Russian ethnic group. We also did not find any association of CYP1A1 gene with COPD in ethnic groups of Bashkortostan. PMID- 18389618 TI - [Influence of polymorphism of immune defense modifier genes on celiac disease development and various clinical features of disease in Tomsk population]. AB - The results of investigation of influence of immune defense modifier genes polymorphism: IL1B (+3953A1/A2), IL1RN (VNTR), IL4A (3'-UTR G/C), IL4RA (I50V), IL12B (1188A/C) and VDR (F/f and B/b) on celiac disease development and various clinical features of disease are presented. The study was performed in 49 families with proband affected by celiac disease (139 people) and 129 unaffected controls of Russian ethnicity from Tomsk. No associations were shown between these alleles and celiac disease by case-control study. However, in family-based investigation, the association was detected for 3'-UTR G/C polymorphism of IL4 gene (p = 0.024). Furthermore, for this polymorphic variant the associations with atypical form of the disease was shown (p = 0.001), as well as with osteopenic (p = 0.039) and thyriopatic (p = 0.042) complications of celiac disease. Association with clinical course of the disease (typical form) was obtained for 150V polymorphism of IL4RA gene and for F/f polymorphism of VDR gene (p = 0.001 and p = 0.009, respectively). Thus, in this investigation it was detected that the associations with the studied phenotypes were found mainly for polymorphic variants of Th2-immunity genes. PMID- 18389619 TI - [Characterization and expression analysis of two cotton genes encoding putative UDP-Glycosyltransferases]. AB - UDP-Glycosyltransferases (UGT) are a large family of enzymes, which catalyze the transfer of a sugar from an activated sugar donor to an acceptor molecule. Both in plant and in mammalian, they are important in maintenance of cellular homeostasis. In this study, two genes (designated GhUGT1 and GhUGT2, respectively) encoding putative UGT were isolated from cotton fiber cDNA library. The deduced proteins contain the signature sequences of plant UGTs in the C terminal region. The GhUGT1 gene encodes a polypeptide of 457 amino acids, and displays homology at amino acid levels with the known glucosyltransferase genes. Sequence analysis revealed that the GhUGT2 merely encodes a small protein, as there is a nucleotide substitution that results in formation of a stop codon in its open-reading frame. Real-time RT-PCR analysis revealed that the expression of GhUGT1 is higher in the fast growth tissues, such as in fibers and roots. GhUGT2 has also higher expression in roots, but with lower expression levels in fibers and other tissues. The result also showed that the expression of GhUGT1 is higher than GhUGT2. Further study showed that GhUGT1 and GhUGT2 expressions are regulated under osmotic stress, suggesting they may be involved in plants responding to osmotic stress. PMID- 18389620 TI - [Cloning, nucleotide sequence and tissue expression profile of three novel porcine genes--RHOB, RHOG and PRAF1]. AB - The complete coding sequences of three porcine genes Rho-related GTP-binding proteins RHOB and RHOG and Prenylated Rab acceptor protein 1 (PRAF1) were amplified using the reverse transcriptase polymerase chain reaction based on the sequence information of the mouse or other mammals and referenced highly homologous pig ESTs. The nucleotide sequences of these three genes revealed that porcine RHOB gene encodes a protein of 196 amino acids that contains the conserved putative RhoA-like domain and has high homology with the RHOB precursor of human, rat and mouse (100%).The porcine RHOG gene encodes a protein of 191 amino acids that contains the conserved putative RhoG domain and has high homology with the RhoG precursor (RHOG) of human, mouse and Cricetus cricetus (98%). The porcine PRAF1 gene encodes a protein of 185 amino acids that contains the conserved putative PRA1 domain and has high homology with the PRAF1 of dog (97%), cattle (97%), human (96%), rat (95%) and mouse (95%). The tissue expression analysis indicated swine RHOB gene was moderately expressed in lung, weakly in fat, spleen, kidney, and almost not expressed in small intestine, large intestine, liver, muscle. The swine RHOG gene was over-expressed in small intestine, large intestine, liver, and muscle, moderately expressed in kidney, weakly in spleen, and almost not expressed in fat and lung. The swine PRAF1 gene was over-expressed in fat and spleen, moderately in lung and kidney, weakly in small intestine and large intestine, and almost not expressed in liver and muscle. PMID- 18389621 TI - [Sequences variation and classification of B-hordein genes in hull-less barley from Qinghai-Tibet Plateau]. AB - The goal of this study is to understand the evolution relationship of the members of B-hordein gene family in hull-less barley by analysis of their structure and to explore their utility in grain quality improvement. Six copies of B-hordein gene (Hn1-Hn3, Hn7-Hn9) were cloned from six hull-less barley cultivars collected from Qinghai-Tibet Plateau and molecularly characterized. Comparison of their predicted polypeptide sequences with the published suggested that they all share the same basic protein structures. In addition, we found that the C-terminal end sequences of all B-hordeins shared a similar feature. In the six clones and the other three published (Hn4, Hn5 and Hn6) from hull-less barley, Hn2 and Hn7 contained identical C-terminal end sequence DIMPVDFWH, Hn3, Hn4, Hn5, Hn8 and Hn9 also shared the common sequence DIMPPDFWH, which was similar to that of a B hordein reported previously. Both Hnl and Hn6 exhibited differences in their C terminal end sequences, and they clustered into different subgroups. The B hordeins with identical C-terminal end sequences were clustered into a same subgroup, so we believe that B-hordein gene subfamilies possibly can be classified on the basis of the conserved C-terminal end sequences of predicted polypeptide. Phylogenetic analysis also indicated that there is a relatively weak identity between our predicted B-hordeins and those reported from H. chilense and H. brevisubulatum. All of our nine predicted B-hordeins were clustered together and other B-hordeins formed another cluster. The possible use of these genes in relation to the barley quality is discussed. PMID- 18389622 TI - [Inactivation of the VHL gene in sporadic clear cell renal cancer]. AB - Renal cell carcinoma is the most common variant of the kidney cancer, which accounts approximately 75% patients with this disease. The majority of those tumors are characterized by inactivation of the VHL gene suppressor as a result of mutations, allelic deletions and/or methylation. We have conducted the complex molecular-genetic analysis of 64 samples obtained from patients with the clear cell renal cancer. VHL mutations were detected by single strand conformation polymorphism and subsequent sequencing, loss of heterozygosity was analyzed using two STR-markers, methylation was tested by methylsensitive polymerase chain reaction. All revealed variations were statistically analyzed in respect to the parameters of primary tumors in various groups of patients. Seventeen VHL somatic mutations were detected, 12 from which were described for the first time. Allelic deletions of VHL were found in 31.6%, and methylation--in 7.8% samples of the renal cancer. As a whole, VHL inactivating events were presented in 46.9% cases of disease, in 51.7% -among renal cancer patients with first stage. We have not observed any association of mutations, loss of heterozygosity and methylation with clinical-pathological parameters of disease. Results of this investigation specify for expediency of further studies of molecular genetics aberrations in the VHL gene. Perhaps, it would promote renal cancer molecular markers evaluation, for example, a determination of suppressor genes methylated in renal cancer. PMID- 18389623 TI - [Molecular characteristic of influenza virus A H5N1 Strains isolated from poultry in Kurgan Region in 2005]. AB - During the latter half of 2005 a widespread outbreak caused by influenza highly pathogenic H5N1 virus among wild and domestic birds occurred in Russia. As pathogenicity level is a polygenic feature and majority of individual genes of influenza A viruses contribute to pathogenicity of influenza viruses to birds, animals and humans. Nucleotide sequencing of the entire genome of influenza H5N1 virus isolates obtained in Kurgan region (Western Siberia) was performed. Structure of viral proteins was analyzed according to the predicted amino acid sequences. HA receptor-binding site of A/chicken/Kurgan/05/2005 and A/duck/Kurgan/08/2005 strains was typical for avian influenza viruses and contained Glu and Gly at positions 226 and 228, respectively. Structure of the cluster of positively charged amino acid residues at the cleavage site was identical for all isolates: QGERRRKKR. According to the data of neuraminidase structure analysis NA of the H5N1 isolates tested was suggested to belong to Z genotype. Amino acid residues typical for birds were revealed in 30 out of 32 positions of M1, M2, NP, PA and PB2 proteins determining host range specificity. One strain isolated in Kurgan contained lysine in position 627 of PB2 protein. Kurgan isolates was shown to have remantadine-sensitive genotype. Glutamic acid was found at position 92 of NS1 protein in both strains indicating virus resistance to interferon. Phylogenetic analyses allowed relating Kurgan isolates to subclade II of clade II of highly pathogenic H5N1 influenza viruses. PMID- 18389624 TI - [Comparative analysis of mitochondrial and nuclear DNA topoisomerase I from maize]. AB - We conducted a comparative study of the properties of topoisomerase I isolated from maize nuclei and mitochondria. We found that nuclear and mitochondrial enzymes possess different ability to bind single stranded DNA. Study of the enzyme activity dependence on Mg2+ demonstrated an absolute dependence of the mitochondrial topoisomerase activity. Contrary, nuclear enzyme activity was not absolutely dependent but stimulated by the magnesium cation. Mitochondrial topoisomerase formed covalent bond with the 5'-end of the cleaved DNA what is unique property of prokaryotic topoisomerase I. Nuclear enzyme bound covalently to the 3'-end like all eukaryotic topoisomerases I. The search through databases revealed genes which could encode mitochondrial topoisomerase I in the genomes of higher plants. Using both cDNA sequencing and in silico methods we demonstrated an existence of the ortholog gene in the maize genome. This gene shares significant homology with prokaryotic topoisomerase I genes that may explain differences in the properties of the mitochondrial and nuclear enzyme. Data obtained is of a significant interest both from the point of view of plant organelle evolution and mitochondrial genome expression mechanisms study. PMID- 18389625 TI - [Frequent allelic losses in tumor-associated stromal cells and tumor epitelium of prostate cancer]. AB - It has become increasingly clear that tumor microenvironment plays a critical role in carcinogenesis. Accumulation of genetic alterations is typical not only for cancer epithelial cells but tumor-associated fibroblasts as well. Tumor epithelia, tumor-associated stroma from prostatectomy specimens of patients with prostate cancer and cells from prostatic intraepithelial neoplasia (PIN) and adjacent stroma from males with PIN were isolated by using laser capture microdissection. Microsatellite allelotyping was evaluated using 4 highly polymorphic markers for chromosomal regions 8p22, 16q23-24 and 13q14. Incidences of alterations (loss of heterozygosity or allelic imbalance) were 48% for region 8p22, 72% for 16q23 and 37% for 13q14. The LOH frequencies in tumor-associated stroma cells were very similar. Alterations at chromosome 13q were significantly associated with advanced tumor stage, whereas AI at 16q was also associated with high Gleason score and lymph node metastasis. We find some incidences of allelic imbalance in premalignant lesions in epithelial (16-27%) and stromal (7-22%) components. Our results show that the frequencies of genetic aberrations are as high in stromal cells as in tumor cells. PMID- 18389627 TI - [Transcription factor YY1 participates in activation transcription of the human ribosomal protein L11 gene]. AB - Ribosomal protein L11 plays important role in ribosome, being involved in several steps in protein synthesis and also activates p53-dependent cell cycle arrest. Changes in the rpL11 levels might be implicated in cell cycle control and carcinogenesis. Therefore, the mechanism of regulation of rpL11 expression has increasing importance. Article presents research results of interaction of promotor elements of gene HRPL11 with proteins of nuclear extracts of cells of a various cell origin. Use oligonucleotide competitors containing known transcription factor-binding sites, and also polyclonal antibodies has shown, that transcription factor YY1 participates in regulation of a transcription of gene HRPL11 in all investigated cellular lines. Our data obtained from comparison of protein binding profiles using nuclear extracts from rapidly growth cells, normal cell lines and serum deprivation repressed cell allows us to consider of transcription factor YY1 as activator of HRPL11 gene transcription. PMID- 18389626 TI - [Reversal of multidrug resistance of gastric cancer cells by down-regulation of CIAPIN1 with CIAPIN1 siRNA]. AB - The overexpression of a new cytokine-induced apoptosis inhibitor 1 (CIAPIN1) gene has been shown previously to promote a multidrug resistant phenotype in gastric cancer cells through the upregulation of MDR1 and MRP1. In the present study, we constructed the siRNA eukaryotic expression vectors of CIAPIN1 and transfected them into SGC7901/VCR cells to examine whether the down regulation of CIAPIN1 increased cell sensitivity towards chemotherapeutic drugs. After transfection, the expression of CIAPIN1 was dramatically decreased in CIAPIN1 siRNA transfectants compared with that in parental cells and empty vector control cells. The down-regulation of CIAPIN1 significantly enhanced the sensitivity of SGC7901/VCR cells to vincristine (VCR), adriamycin (ADR) and etoposide (VP-16), but not to 5-fluorouracil (5-FU) and cisplatin (CDDP). Cell capacity to efflux adriamycin decreased markedly in CIAPIN1 siRNA transfectants, and correlation between CIAPIN1 down regulation and decreased MDR1 transcriptional activity were observed. CIAPIN1 siRNA could significantly down regulate the expression of Bcl 2, and up-regulate the expression of Bax, but not alter the expression of PTEN in gastric cancer cells. These observations suggested that the siRNA constructs of CIAPIN1 we obtained could effectively down-regulate the expression of CIAPIN1 and reverse the resistant phenotype of gastric cancer cells. The further study of the biological functions of CIAPIN1 may be helpful for understanding the mechanisms of multidrug resistance of gastric cancer and developing possible strategies to treat gastric cancer. PMID- 18389628 TI - [Start codon in the serine proteinase gene from Bacillus intermedius]. AB - The translation initiation site in the extracellular serine subtilisin-like proteinase gene from Bacillus intermedius (aprBi) (AN AY754946) secreting at the stationary growth phase was established. The analysis of aprBi open reading frame revealed three putative translation start sites (TTG, GTG, ATG). Using SignalP online freeware program we have determined the functional activity probability of each of them. To identify the translation start point the modified subtilisin like protease genes carrying nucleotide replacements in supposed start codons were developed using oligonucleotide-directed mutagenesis. We have investigated the expression of these genetic constructions in protease-deficient strain B. subtilis AJ73. According our results it was concluded that the translation in aprBi gene starts from GTG kodon. PMID- 18389629 TI - [Yeast chaperone Hspl04 regulates gene expression on the posttranscriptional level]. AB - Yeast chaperon Hsp104 is known as a protein which is able to dissociate aggregates of the heat damaged proteins and prion aggregates into smaller pieces or monomers. In our work the effects of Hsp104 on the PrP-GFP and GFP proteins have been analyzed. The PrP-GFP protein forms the high molecular weight aggregates, whereas GFP is unable to aggregate in yeast cell. We have shown that Hsp104 regulates the amount of PrP-GFP and GFP in yeast cells and direction of chaperone action depends on promoter controlling production of these proteins. The overproduction of Hsp104 increases the amount of PrP-GFP and GFP proteins when the corresponding genes are under control of CUP1 promoter. In contrast, the overproduction of Hsp104 decreases the amount of PrP-GFP and GFP is case of their expression under control of GPD promoter. The effects of Hspl04 are not related with any changes in mRNA content of the genes under investigation and with ability of the proteins to form aggregates. Thus, the functions of this chaperon are not restricted by dissociation of the protein aggregates. Our data show that Hsp104 regulates the gene expression on the posttranscriptional level. PMID- 18389630 TI - [Dimerization of short RNA fragments from avian leukosis virus as revealed with 2 aminopurine fluorescence]. AB - The dimerization of genomic retroviral RNA is well studied for several groups of viruses, the dimerization of human immunodeficiency (HIV) RNA being investigated in more detail. Regions of dimerization apparently involve the short sequences RNA which are directly responsible for the formation of two type dimers: kissing loop-loop (KD) and linear (LD). The 5'-end sequences from RNA avian viruses, where the dimers are basically formed, considerably differ from those of HIV. However, as it was described earlier, the mechanism of dimerization of RNA from human immunodeficiency and from avian leukosis viruses are identical. The fluorescence of adenine analogue 2-aminopurine (2-AP) incorporated into loop sequence of short fragments RNA ALV was used for analysis of dimers formation. Using the temperature dependence of fluorescence intensity 2-AP we have determined RNA melting temperature under various conditions for KD RNA ALV formed by two strands. Effects of magnesium and aminoglycoside antibiotic paromomycin on stabilization of kissing loop-loop dimer RNA have been studied. Under the experimental conditions KD RNA ALV was found to have the stability at the magnesium concentration higher than 1 mM and at paromomycin concentration higher than 2.5 mkM. PMID- 18389631 TI - [A simple kinetic model for dynein oscillatory activity]. AB - A kinetic model for dynein, a molecular motor, complexed with microtubule fragments, is considered. The model explains the experimental observations of oscillatory movements in surprisingly simple axonemal fragments perfused by the ATP solution. The model explains at first time the oscillatory dynein activity as a phenomenon induced by two dynein heads cooperative interaction in the axoneme. The oscillation form, frequency, and amplitude, observed for the model, are close to these experimental characteristics. Kinetic parameters, used in the model, are close to the known experimental parameters. PMID- 18389632 TI - [Orphan genes: function, evolution, composition]. AB - The origin of orphan genes and the function they perform remain an enigmatic problem that modern molecular biology has to solve. The recently developed PHOG database helped to shed light on some aspects of the evolution of these genes. Fast evolution is probably the main factor that shapes this evolutionary process. The speed of molecular evolution reflects the degree of gene conservation, though exceptions from this rule also contribute to the dynamic process of genome evolution during speciation events. The existence of the great number of orphan genes is an artifact of insufficient sequencing. If the DNA of all organisms living on Earth were sequences, then the number of orphan genes will be greatly reduced, giving the way to the genes specific for particular taxonomic groups. PMID- 18389633 TI - [Statistical analysis of DNA sequences nearby splicing sites]. AB - Recognition of coding regions within eukaryotic genomes is one of oldest but yet not solved problems of bioinformatics. New high-accuracy methods of splicing sites recognition are needed to solve this problem. A question of current interest is to identify specific features of nucleotide sequences nearby splicing sites and recognize sites in sequence context. We performed a statistical analysis of human genes fragment database and revealed some characteristics of nucleotide sequences in splicing sites neighborhood. Frequencies of all nucleotides and dinucleotides in splicing sites environment were computed and nucleotides and dinucleotides with extremely high?low occurrences were identified. Statistical information obtained in this work can be used in further development of the methods of splicing sites annotation and exon-intron structure recognition. PMID- 18389634 TI - [siRNAs with high specificity to the target: a systematic design by CRM algorithm]. AB - 'Off-target' silencing effect hinders the development of siRNA-based therapeutic and research applications. Common solution to this problem is an employment of the BLAST that may miss significant alignments or an exhaustive Smith-Waterman algorithm that is very time-consuming. We have developed a Comprehensive Redundancy Minimizer (CRM) approach for mapping all unique sequences ("targets") 9-to-15 nt in size within large sets of sequences (e.g. transcriptomes). CRM outputs a list of potential siRNA candidates for every transcript of the particular species. These candidates could be further analyzed by traditional "set-of-rules" types of siRNA designing tools. For human, 91% of transcripts are covered by candidate siRNAs with kernel targets of N = 15. We tested our approach on the collection of previously described experimentally assessed siRNAs and found that the correlation between efficacy and presence in CRM-approved set is significant (r = 0.215, p-value = 0.0001). An interactive database that contains a precompiled set of all human siRNA candidates with minimized redundancy is available at http://129.174.194.243. Application of the CRM-based filtering minimizes potential "off-target" silencing effects and could improve routine siRNA applications. PMID- 18389636 TI - [Potential signals for transcription termination of integrated HPV DNA: analysis in silico]. PMID- 18389635 TI - [Inhibition of agrobacterial oncogenes expression by means of antisense RNA]. AB - Plant's infection with soil bacteria Agrobacterium tumefaciens lead to tumour formation, so called crown galls. The reason of tumorigenesis is integration of agrobacterial genes for phytohormone synthesis auxins and cytokinins in plant genome, the most important of them are iaaM and ipt. Obtaining of transgenic plants able to inhibit these genes expression, creates conditions for producing of plants resistant to crown gall disease. With this purpose single and double transformants of tobacco plants with antisense copies of iaaM and ipt genes under the control of single and double promoters of 35S RNA of cauliflower mosaic virus (CaMV 35S and CaMV 35SS) were produced. Infection with virulentA. tumefaciens strains C58 (pTiC58) and A6 (pTiA6) of all types transgenic plants with antisense oncogenes copies showed essential but incomplete inhibition of these genes expression. After agrobacterial transformations of transgenic plants only "weakened" tumours of various morphology, able to regenerate the whole plants, were formed. The analysis data of inhibition of iaaM and ipt genes expression in formed tumour cells were presented. The results indicate perspective RNA interference strategy for producing of plants resistant to agrobacterial crown gall disease. PMID- 18389637 TI - [Redox modification and phosphorylation modulate activity of mitochondrial DNA binding protein complex which is specific for maize Coxi promo]. PMID- 18389638 TI - [Sequencing of 5'-region of Lngfr gene associated with stress-induced arterial hypertension in the ISIAH rats]. PMID- 18389639 TI - Reconciling clinical experience with evidence-based knowledge in suicide prevention policy and practice. PMID- 18389640 TI - The changing gender ratio in occurrence of deliberate self-harm across the lifecycle. AB - BACKGROUND: Overall gender ratios are often quoted in studies of deliberate self harm (DSH) patients, almost always with higher rates in females than males. Reporting a ratio across all ages may conceal important variations in the gender ratio across the life cycle. METHOD: Analysis was done of the gender ratio by age groups in rates of DSH in a consecutive sample of DSH patients presenting to a general hospital over a 10-year period. The patients were identified through a well-established monitoring system. RESULTS: The study sample included 2,189 female and 1,439 male patients. While the overall gender rate ratio was 1.5 females to each male, the ratio varied considerably by age group: 8:1 in 10-14 year-olds, 3.1:1 in 15-19-year-olds, 1.6:1 in 20-24-year-olds, approximately 1.3:1 in 25-49-year-olds, and 0.8:1 in people aged 50 years and over. CONCLUSION: Statements about overall gender ratios for DSH conceal important changes in the ratio across the life cycle. These changes probably reflect differences in development and problems faced in adolescence, changes in motivation for DSH with age, and the closer resemblance of DSH to suicide in older age groups. PMID- 18389642 TI - Self-reported suicidal behavior and attitudes toward suicide and suicide prevention among psychology students in Ghana, Uganda, and Norway. AB - Self-reported suicidal behavior and attitudes toward suicide in psychology students are reported and compared in Ghana, Uganda, and Norway. Small differences only were found in own suicidal behavior. However, experience of suicidal behavior in the surroundings was more common in Uganda than in Ghana and Norway. Although differences were found between the three countries in attitudes toward suicide, which emphasizes the need for culture-sensitive research and prevention, many of the differences were not as big as expected. The most pronounced difference was that the Norwegian students were more reluctant to take a stand on these questions compared to their African counterparts. Some differences were also found between the two African countries. The implications of the results for suicide prevention in Africa are discussed. PMID- 18389641 TI - Social modeling in the transmission of suicidality. AB - Evidence from twin, adoption, and family studies suggests that there is strong aggregation of suicidal behaviors in some families. By comparison, the role of social modeling through peers has yet to be convincingly established. This paper uses data from four large studies (the WHO/EURO Multicentre Study on Suicidal Behaviour, the WHO/SUPRE-MISS, the CASE study, and the Queensland Suicide Register) to compare the effects of exposure to fatal and nonfatal suicidal behavior in family members and nonfamilial associates on the subsequent suicidal behavior of male and female respondents of different ages. Across all studies, we found that prior suicidal behaviors among respondents' social groups were more important predictors of suicidal behavior in the respondents themselves than previous research had indicated. Community-based suicide attempters in the WHO SUPRE-MISS had higher rates of exposure to prior suicide in nonfamilial associates than in family members. In an adolescent population, exposure to prior fatal suicidal behavior did not predict deliberate self-harm when exposure to nonfatal suicidal behavior (both familial and social) were controlled for, but exposure to nonfatal suicidal behaviors in family and friends was predictive of deliberate self-harm and suicide ideation, even after controlling for exposure to fatal suicidal behavior. The potential impact of "containment" of information regarding suicidal behaviors as a prevention initiative is discussed, in light of information behavior principles of social marketing. PMID- 18389643 TI - Deliberate self-harm: characteristics of patients presenting to a tertiary care hospital in Karachi, Pakistan. AB - INTRODUCTION: Previous deliberate self-harm (DSH) is the strongest predictor of suicide. Although several studies exist in other countries, characteristics of DSH have not been well-studied in Pakistan. AIMS: To identify characteristics of deliberate self-harm in a hospital population, building on previous work carried out on this subject at this center. METHODS: 284 cases admitted to the Aga Khan University Hospital, Karachi, between January 1997 and August 2003 were studied by analyzing medical records. Information was collected regarding demographics, family history, personal history, suicidal ideation, current stressors, the act of DSH and its management. RESULTS: The most common age group was 21-25 years of age. The majority were females (60.3%). 95.1% of patients used self-poisoning as a method of deliberate self-harm. 47.3% used benzodiazepines for self-poisoning. Precipitating factors included conflict with family, marital problems, chronic illnesses, and unemployment. Comorbid psychiatric disorders were present in 76.4% patients. DISCUSSION: Despite the difference in socio-cultural background of our patients, many of our results were found to be consistent with Western data. Our findings suggest that problem-solving therapy, stress management and improved mental health care may be effective interventions in preventing DSH. Prescription of medications should be regulated. Further studies are suggested at the community level. PMID- 18389644 TI - The impact of suicide on the family. AB - Little research has examined the consequences of a suicide for social or family networks. Because suicide occurs within families, the focus on the aftermath of suicide within families is an important next step to determine exactly how to help survivors. In this article, we review and summarize the research on the impact of suicide on individuals within families and on family and social networks. We begin with a discussion of family changes following suicide. Next, we discuss the effects of suicide on social networks overall and responses of children and the elderly to a suicide in the family. Finally, we identify key issues that remain to be resolved in family survivor research and make recommendations for future studies. PMID- 18389645 TI - Surviving the suicides of significant others: a case study. AB - Psychological pain has been reported to be major contributing factor for completed suicide, and classical suicidology based its wisdom on this construct. In this paper we report the case of a patient whose family experienced a high number of suicides apparently due to an escalation of psychological pain in the family members after the successive suicides. We also briefly discuss the problem of helping survivors after the suicide of loved one. PMID- 18389646 TI - Suicide among indigenous peoples: the usefulness of the Human Relations Area Files. AB - The present note describes the resource for studying suicide in indigenous peoples provided by the Human Relations Area Files and describes three ways in which these files can illuminate suicidal behavior: testing theories of suicide, exploring the linguistics of suicide, and challenging myths about suicide. PMID- 18389647 TI - Remarks by Mary McAleese, President of Ireland made at the International Association of Suicide Prevention XXIV Biennial Conference, August 31, 2007, Irish National Events Centre, Killarney, Co. Kerry, Ireland. AB - Reducing suicide rates requires a collective, concerted effort from all groups in society: health, social services, other professionals, communities and community leaders, voluntary and statutory agencies and organizations, parents, friends, neighbors, and individuals. It also requires the careful nurturing of a culture in which people in psychological distress don't hesitate to seek help from family, friends, health professionals and community leaders, a culture that recognizes the signs and signals of distress and is willing to help, and that focuses early in life on developing good coping skills and avoiding harmful practices; in short, a sensitive culture that cherishes human courtesy and takes responsibility for our own mental health and the mental health of others. Suicide occurs in many diverse contexts but there are recurring patterns and elements that research is revealing to us. The more we are learning, the better we are able to devise strategies and take action. We already know, for example, that alcohol abuse plays a very significant role in suicide and features prominently in youth suicide. Research has also established that cannabis increases the risk of depression and psychosis and that it reduces normal inhibitions against suicide. Since marijuana is the illegal drug most used by those under25 years of age, a critical factor in suicide prevention must be to inform young people as to its danger and counter the erroneous impression that it is a benign substance. We know that bullying sometimes features in the story of suicide. Our young people need to know just how damaging, just how unacceptable, bullying is. Education and access to support structures - whether at work, school, or home - are an essential part of protecting the victim of bullying not just from the bully but from their own downward spiral into suicidal thoughts. A less discussed but nonetheless important issue worth raising is the issue of dealing with sexual identity. Although Ireland is making considerable progress in developing a culture of genuine equality, recognition, and acceptance of gay men and women, there is still an undercurrent of both bias and hostility which young gay people must find deeply hurtful and inhibiting. For them, homosexuality is a discovery, not a decision, and for many it is a discovery which is made against a backdrop where, within their immediate circle of family and friends as well as the wider society, they have long encountered antigay attitudes which will do little to help them deal openly and healthily with their own sexuality. PMID- 18389648 TI - Taking on the formidable middle. PMID- 18389649 TI - The forgotten flu. PMID- 18389650 TI - Research not just for research's sake. PMID- 18389651 TI - The 3Cs for recovery services. PMID- 18389652 TI - Learning from top performers. PMID- 18389653 TI - Considering the possibilities of multiple specimen drug testing. PMID- 18389654 TI - The handwriting was on the wall. PMID- 18389655 TI - A Rx to improve medication safety. PMID- 18389656 TI - Prescribers can't completely eliminate paper--yet. PMID- 18389657 TI - Are you ready for e-prescribing? PMID- 18389658 TI - Finding solutions to a gray future. PMID- 18389659 TI - Collaboration. PMID- 18389660 TI - Lusitropic effects of dobutamine in young and aged mice in vivo. AB - Aged individuals have impaired diastolic relaxation-lusitropic function. Dobutamine, a selective B1-adrenergic agonist, is used to augment systolic cardiac function at the termination of cardiopulmonary bypass (CPB). However, our question is whether dobutamine will also enhance the lusitropic function in the aged individual. The myocyte mechanism for the rate of ventricular relaxation is dependent on the velocity of calcium removal from the myocyte contractile elements by sarcoplasmic reticulum (SR) Ca(2+)-ATPase (SERCA2a), which is regulated by an inhibitory protein, phospholamban (PLB). Ventricular tissues harvested from young (4 month) and aged (20 months) mice were analyzed to compare the protein levels of SERCA2a and PLB with immunoblot and gene expression for PLB with reverse-transcriptase-polymerase chain reaction. The molecular analyses were compared with in vivo left ventricular function in the young and old mice before and during an intravenous infusion of dobutamine (5 microg/kg/min). The SERCA2a levels were not different between the groups; however, there was a 2-fold increase in PLB in the aged group compared with the young group (p < .05). The gene expression for PLB was increased by 5-fold in the aged group compared with the young group (p < .01). There were significant differences between the young and aged groups related to the lusitropic parameters, tau and dP/dt(min), and dobutamine infusion increased these parameters in the aged group to that of the young group. This report supports the concept that altered PLB levels correspond with the respective lusitropic function and that dobutamine administration in the aged group increased lusitropic function that was comparable with the young group. Because the patient population requiring CPB is aging, these data suggest that the use of dobutamine at the terminal phase of CPB is warranted to increase systolic and diastolic function. PMID- 18389661 TI - Cardiopulmonary bypass recommendations in adults: the northern New England experience. AB - Using a regional cardiopulmonary bypass (CPB) registry, we compared the practice of CPB at eight northern New England institutions to recently published recommendations. We examined CPB practice among 3597 adult patients undergoing isolated coronary artery bypass grafting surgery from January 2004 to June 2005. Registry variables were used to compare regional CPB practice to recommendations on topics of neurologic protection (pH management, avoidance of hyperthermia, minimizing return of pericardial suction blood, aortic assessment, arterial line filtration), maintenance of euglycemia, reduction of hemodilution, and attenuation of the inflammatory response. We report overall regional practice (regional minimum, maximum). All centers used alpha-stat pH management and arterial line filters. Avoidance of hyperthermia (temperature < 37degrees C) was achieved during 23.4% of procedures (regional minimum, 1.5%; maximum, 83.2%). Minimizing return of pericardial suction blood was achieved in 23.7% of cases (0.7%, 93.6%). Aortic assessment was performed during 45.7% of procedures (1.3%, 98.9%). Maintenance of euglycemia (< 200 mg/dL) was accomplished in 82.7% (57.1%, 97.9%) of cases. Hemodilution (hematocrit < 23% on CPB) was lower for men 32.4% (20.6%, 52.3%) than women 77.9% (64.7% 88.9%). Men were less likely to receive red blood cell transfusions in the operating room (11.0%; 1.8%, 20.9%) than women (54.6%; 30.1%, 70.6%). In an effort to attenuate the inflammatory response, surface coated circuits were used in 83.3% of procedures (8.8%, 100%). During this time, gaps existed between regional CPB practice and recently published recommendations. We continue to prospectively measure CPB practice relating to these recommendations to monitor and improve the care provided to our patients. PMID- 18389662 TI - Arterial line filters ranked for gaseous micro-emboli separation performance: an in vitro study. AB - Arterial line filters (ALFs) are arguably the most important component in the cardiopulmonary bypass circuit to protect the patient from gaseous macro- and micro-emboli (GME) originating in the perfusion circuit. The GME separating ability of 10 ALFs was ranked according to seven performance criteria. Ten ALFs rated between 20 and 43 microm were evaluated for flow resistance, the count, size, and volume of GME passed after a 10-mL room air bolus, and the ability to separate a high-count, 10- to 200-microm flowing distribution of GME. The Luna Innovations EDAC emboli detector was used to size, count, and sort GME. Three test trials were conducted for 3 each of the 10 filters. Performance criteria were correlated by regression analysis, statistically compared using analysis of variance, or ranked using non-parametric tests. Significance was set at 0.05. Weighting all seven test parameters equally, the most effective ALFs were the Cobe 21 and Gish 25-microm filters. The Pall LG-6 ranked more efficient than the Medtronic 20 and Dideco 27-microm filters. The Cobe 43, Terumo 40, Medtronic 38, Terumo 37, and Gish 40-microm filters were less effective as a group compared with the other filters. For the 10 filters, blood flow resistance was not correlated to rated pore size. Generally, the smaller the pore rating, the higher the GME separation ability rank, except for the leuko-reduction filter, which performed more effectively than other large pore filters. KEYWORDS: arterial line filter, gaseous microemboli, in vitro test. PMID- 18389663 TI - Use of leukocyte-depleting filters during cardiac surgery with cardiopulmonary bypass: a review. AB - Numerous researchers and clinicians have shown that cardiopulmonary bypass (CPB) plays a large role in the initiation of the systemic inflammatory response during cardiac surgery. The activation of leukocytes during this process has been implicated as one of the major contributors to multi-organ dysfunction experienced by some patients after cardiac surgery. Thus, in an attempt to attenuate the systemic inflammatory response and to reduce the amount of activated leukocytes from the systemic circulation during CPB, leukocyte depleting filters were developed in the early 1990s. Since the clinical introduction of these filters into the CPB circuit, several articles have been published evaluating the effectiveness of leukocyte filtration; however, the results have been conflicting. This article will review some of the most recent literature, approximately 40 papers published within the past 10 years, regarding the use of leukocyte-depleting filters during CPB and its effectiveness in preserving organ function. In addition, the effect of different filtration strategies and the effectiveness of the filter to attenuate the systemic inflammatory response in combination with other mechanical and pharmaceutical strategies will be reviewed. PMID- 18389664 TI - Factors that influence the ability to perform autologous priming. AB - The purpose of this study was to determine which factors impact the ability to perform autologous priming (AP) of the extracorporeal circuit. Second, the effects of differential AP on transfusion and volume requirements were evaluated. After institutional review board approval, demographic, operative, volumetric, and transfusion data were prospectively collected on 100 adult patients undergoing cardiopulmonary bypass (CPB). Two analyses were conducted: AP Taken and percent AP Given. For each analysis, three groups were created based on standard distribution. Group A included patients within less than mean--1 SD (< or = 500 mL AP Taken or > or = 90% AP Given back), group B included patients within mean +/- 1 SD (501-1299 mL AP Taken or 11%-89% AP Given back), and group C included patients greater than mean + 1 SD (> or = 1300 mL AP Taken or < or = 10% AP given back). Weight, pre-CPB hematocrit, clinical severity, and pre-CPB volume balance did not differ between the groups. Significant differences existed in AP Taken and percent AP Given between individual perfusionists. More AP was given back with higher urine output (group A: 846 +/- 700 mL, group B: 613 +/- 414 mL, group C: 384 +/- 272 mL; p = .004), more autotransfusion [group A: 0 (0,1300 mL), group B: 0 (0,500 mL), group C: 0 (0,250 mL); p = .008], and less AP Taken [group A: 800 (0,1300 mL), group B: 1000 (200,1600 mL), group C: 1000 (800,1600 mL); p = .001]. When more AP was taken, CPB hematocrit was higher (group A: 22.3% +/- 4.8%, group B: 25.6% +/- 4.7%, group C: 26.6% +/- 4.3%; p = .032), and fewer patients received red blood cells (group A: 64.3%, group B: 28.3%, group C:14.3%; p = .017). Some perfusionists were able to remove more AP before CPB. When more AP was taken, CPB hematocrit was higher, fewer patients received a transfusion, and less AP was given back. More AP was also given back with higher urine output and higher blood loss to the autotransfusion device. PMID- 18389665 TI - Non-activated autologous platelet-rich plasma for the prevention of inguinal wound-related complications after endovascular repair of abdominal aortic aneurysms. AB - The endovascular repair (EVAR) of abdominal aortic aneurysms (AAAs) usually involves the surgical exposure and catheterization of the femoral arteries. Several inguinal surgical wound-related complications have been reported postoperatively. The aim of this report was to evaluate the safety and efficacy of intraoperative application of autologous platelet-rich plasma (PRP) for the prevention of wound-related complications in AAA EVAR. The authors conducted a patient- and assessor-blinded controlled trial involving 100 subjects undergoing EVAR of an AAA. PRP was produced using an autologous platelet separator and was applied, without prior thrombin activation, in 50 patients eligible for inclusion. The results were compared with a control group of 50 patients who underwent AAA EVAR within the same time period. The primary outcome was the difference in postoperative hospital stay. Secondary outcomes included subjective assessment of wound healing and wound-related complications. Age, sex, and other comorbidities related to wound healing were not significantly different between cases and controls. One patient treated with PRP developed a unilateral wound infection with lymphorrhea, and two patients developed a bi-lateral superficial infection. Twelve patients within the control group developed a wound-related complication. The postoperative hospitalization was significantly lower in the PRP group. The overall surgical wound-related complications rate was also significantly lower in the PRP group. Application of non-thrombin-activated PRP seems to prevent major postoperative wound-related complications (p = .026) and shorten postoperative hospital stay duration after femoral artery exposure and catheterization for AAA EVAR (mean, 4.48 +/- 0.48 vs. 6.14 +/- 0.39 days). PMID- 18389666 TI - Evaluation of the i-STAT point-of-care analyzer in critically ill adult patients. AB - Point-of-care analyzers may benefit therapeutic decision making by reducing turn around-time for samples. This is especially true when biochemical parameters exceed the clinical reference range, in which acute and effective treatment is essential. We therefore evaluated the analytical performance of the i-STAT point of-care analyzer in two critically ill adult patient populations. During a 3 month period, 48 blood samples from patients undergoing cardiac surgery with cardiopulmonary bypass (CPB) and 42 blood samples from non-cardiac patients who needed intensive care treatment were analyzed on both the i-STAT analyzer (CPB and non-CPB mode, respectively) and our laboratory analyzers (RapidLab 865/Sysmex XE-2100 instrument). The agreement analysis for quantitative data was used to compare i-STAT to RapidLab for blood gas/electrolytes and for hematocrit with the Sysmex instrument. Point-of-care electrolytes and blood gases had constant deviation, except for pH, pO2, and hematocrit. A clear linear trend in deviation of i-STAT from RapidLab was noticed for pH during CPB (r = 0.32, p = .03) and for pO2 > 10 kPa during CPB (r = -0.59, p < .0001 when 10 .05) and last (24.5 +/- 5.9% vs. 25.1 + 5.7%, r = 0.9001, p > .05) bypass hematocrit. Using 65 mL/kg resulted in a larger difference between estimated and measured hematocrits for the first (25.6 +/- 4.5% vs. 24.5 +/- 6.2%, r = 0.6885, p = .030) and last (23.8 +/- 3.6% vs. 25.1 +/-5.7%, r = 0.5990, p = .001) bypass hematocrits. Compared with using 65 mL/kg for blood volume, the BVE allowed for a more precise estimated hematocrit during CPB. PMID- 18389668 TI - Use of a new implantable adjustable pulmonary artery banding device: a report of two patients. AB - Pulmonary artery banding (PAB) is a method for reducing pulmonary artery blood flow and pressure by tying a surgical band around the main pulmonary artery (MPA). Originally, the procedure was primarily performed to palliate intracardiac left to right shunts for ventricular septal defects (VSDs). Currently, the use of PAB has expanded to allow it to be used in ventricular training, prior to total corrective surgery. At the Rigshospitalet, Copenhagen, Denmark we have been trying a new device. The Flo-Watch PAB implant (EndoArt, Lausanne, Switzerland) is only currently available within certain European countries. The device is a mechanical PAB that allows noninvasive adjustment of the band post implantation, in an outpatient clinic. This has multiple advantages, not only in terms of convenience for the patient, and the reduced risk of multiple surgical interventions, but also from a financial standpoint to the health system. We would like to present our findings and comment on the use of the device in this report. PMID- 18389669 TI - On-line autotransfusion waste calculator. AB - Cell concentrating and washing techniques are widely accepted and believed to be beneficial to cardiac surgery patients. During cell processing, platelets, proteins, and clotting factors are wasted as the plasma is washed away by saline. Beneficial and costly plasma constituents are sacrificed for the sake of removing potentially harmful drugs, debris, and naturally activated cells and chemical mediators. An interactive Microsoft Excel spreadsheet was designed to input patient and autotransfusion system (ATS) reservoir blood values, processed centrifugal bowl data, and hospital allogeneic blood product concentration and cost information. The spreadsheet calculates the number of wasted platelets, grams of protein, and milligrams of fibrinogen. The calculator further estimates the number of units and cost of allogeneic blood products needed to replace the wasted blood components. The simulation allows for variable levels of platelet activation and protein removal during centrifugal cell processing. Specific case scenarios may be simulated with the calculator. If a known volume of residual extracorporeal circuit blood with a known hematocrit, platelet count, and protein concentration is diverted to the ATS reservoir to be processed and washed after bypass, the number of units of fresh frozen plasma, platelet packs, and albumin concentrate needed to replace the wasted proteins and platelets may be calculated. When typical end-bypass patient and blood bank product values are input, the cost to replace the wasted blood components in 1550 mL of residual circuit blood with allogeneic blood products is about US $2097. There are risks and costs associated with replacing the platelets, proteins, and clotting factors wasted during cell washing compared with other techniques such as whole blood ultrafiltration. PMID- 18389670 TI - Rupture of extra-corporeal circuit tubing during cardiopulmonary bypass. AB - Roller pumps are widely used for cardiopulmonary bypass in developing nations by virtue of proven safety during several years of institutional use and cost effectiveness. However, careful adjustment of roller occlusion is needed because they are known to cause spallation, tubing wear, and the occasional incident of rupture of tubing in the extracorporeal circuit. Rupture of polyvinylchloride tubing in the pump raceway during repair of a ventricular septal defect in a 4 year-old child is discussed. The event was managed by exclusion and replacement of the defective tubing during a short period of arrest. Use of an inappropriate boot pump and failure to detect its inclusion in the bypass circuit was a significant departure from protocol. However, because occlusion settings and duration of perfusion were within acceptable limits, a manufacturing flaw could also have contributed to tubing failure, and the event may or may not have been averted by the use of larger tubing. In conclusion, this incident reiterates the need for adherence to established protocol during assembly of the pump and draws attention to the fact that tubing integrity is not a guarantee and vigilance is warranted to handle its failure. PMID- 18389671 TI - Considerations for the use of enteral sedation in pediatric dentistry. AB - Treating an uncooperative, uncontrollable child can be unpleasant for all parties involved. Despite the dentist's best efforts to employ traditional techniques, the behavioral management of challenging pediatric dental patients often requires more than "tell, show, do". Consequently, pre-operative pharmacological intervention may be necessary. Enteral sedation may be the optimal adjunct for the dental treatment of such a challenging patient population. However it must be utilized with caution and is not an appropriate treatment modality for all. This paper will present various considerations for the safe, appropriate and effective use of enteral sedation in contemporary pediatric dentistry. With the strong demand for this service, properly trained practitioners can broaden their practice and provide a win-win scenario for themselves and their patients. PMID- 18389672 TI - Assessment of intranasal midazolam administration with a dose of 0.5 mg/kg in behavior management of uncooperative children. AB - AIM: Intranasal midazolam has been used to induce conscious sedation in children with negative and aggressive behavior The main goal of this study was to determine the effectiveness of intranasal administration of midazolam (with a dose of 0.5 mg/kg) in behavior management of uncooperative children. MATERIALS AND METHODS: Thirty healthy, difficult children of 3-5 years were evaluated. At the beginning of each session, ordinary techniques of behavior management to treat patients were applied. In cases of unsatisfactory responses, intranasal midazolam was immediately employed. To determine the efficacy of the drug, child behavior was evaluated before and after administration of midazolam using Houpt rating scale of general behavior. RESULTS: A statistically significant difference was demonstrated in the patients 'behavior before and after administration of intranasal midazolam. CONCLUSIONS: this drug is effective in sedation and reduction of the anxiety of children under treatment. PMID- 18389673 TI - Root surface characteristics of children teeth with periodontal diseases. AB - The present study examined the root surfaces of teeth from children with or without periodontal diseases and with or without systemic diseases. Light microscopy revealed that when compared to control teeth: teeth with chronic periodontitis had similar radicular histology; teeth from children with leukocyte adhesion deficiency, Down syndrome and aggressive periodontitis had narrower cementum areas; teeth from children with hypophosphatasia showed cementum aplasia. Cementum anomalies may facilitate the establishment and progress of periodontitis in children. PMID- 18389674 TI - Cytotoxicity, histopathological, microbiological and clinical aspects of an endodontic iodoform-based paste used in pediatric dentistry: a review. AB - This review aims at describing and comparing materials commonly used in root canal therapy, the cytotoxicity, histopathological, microbiological and clinical aspects ofa iodoform-based paste (Guedes-Pinto Paste-GPP) used in endodontic treatment of primary teeth. GPP has shown excellent biocompatibility to pulp fibroblasts and mild inflammatory reactions, having been well-tolerated by the periapical and connective tissues. Moreover, GPP bactericidal and bacteriostatic effects against many oral microorganisms were also demonstrated. Regarding clinical trials, the GPP technique has achieved success rates when considering clinical and radiographic examinations. In the face of all the above mentioned results, this paper would like to propose the use of this endodontic material as a root canal filling for primary teeth. PMID- 18389676 TI - Evaluation of residual dentin after conventional and chemomechanical caries removal using SEM. AB - The purpose of this in vitro study was to analyze the residual dentinal surfaces following caries removal using rotatory instruments and two chemomechanical methods (Papacarie and Carisolv), by scanning electron microscopy (SEM). Thirty primary incisors were divided into three groups, according to the caries removal method used, and their residual dentin was examined under SEM (15). After caries removal, 15 of these teeth were restored with Single Bond (3M) adhesive system and Z100 Filtek composite resin (3M). The tags of the replicas were observed under SEM. The chemomechanical caries removal methods (Papacarie and Carisolv) formed an amorphous layer, similar to the smear layer and few exposed dentinal tubules; the conventional caries removal method produced a smooth and regular dentinal surface, with typical smear layer and exposed dentinal tubules. All groups showed abundant tag formation. Scanning electron microscopy analysis revealed a difference between dentin treated with rotatory instruments and that treated with chemomechanical methods in spite of the occurrence of a similar tag formation in both groups. PMID- 18389675 TI - Effect of various mouthwashes on the levels of interleukin-2 and interferon-gamma in chronic gingivitis. AB - The aim of this double blind study was to evaluate the effect of various mouthwashes: Chlorhexidine, Essential oil, Azadirachta indica (Neem) extract, and Povidone iodine on gingival tissue interleukin-2 (IL-2) and interferon-gamma (IFN gamma) levels in patients with chronic gingivitis. A total of 8O patients (42 boys, 38 girls; mean age 16.0 +/- 1.8 years) were included in this study. Patients were randomly assigned into four groups of 20 each: Group I--Azadirachta indica (Neem) extract, Group II--Essential oil, Group III--Povidone iodine, and Group IV--Chlorhexidine. They were instructed to use these mouthwashes for two weeks. Plaque and gingival indices scores, and IL-2 and IFN-gamma levels in the gingival tissues were measured at baseline and after two weeks of mouthwash use. Results showed the reduction of plaque and gingival indices, and IL-2 and IFN gamma level with Chlorhexidine, Essential oil, and Povidone iodine, which were found to be statistically significant. Although Neem reduced the level of plaque and gingival indices, and IL-2 and IFN-gamma to a certain level, it was not statistically significant. Therefore, Chlorhexidine, Essential oil, and Povidone iodine mouthwashes can be used as an adjunct to oral prophylaxis in reducing pro inflammatory cytokines, IL-2 and IFN-gamma in patients with chronic gingivitis. PMID- 18389677 TI - Distribution of Streptococcus mutans and Streptococcus sobrinus in saliva of Mexican preschool caries-free and caries-active children by microbial and molecular (PCR) assays. AB - The aim of the study was to investigate the distribution of mutans streptococci (MS) infection of caries-free (CF) and caries-active (CA) preschool Mexican children by microbial and molecular assays. Eighty preschool children were divided into two groups, 40 CF and 40 CA children. Saliva samples were inoculated onto MSB to identify CFU and DNA extractions were tested by PCR. Our results indicated that there was no statistical difference (p > 0.05) between groups either in age, weight, height or sex. S. sobrinus was detected by PCR twice as much in the CA group, the difference being statistically significant (p < 0.05). dmfs index was positive correlated with S. mutans (r = 0.2941, p = 0.0081), S. sobrinus (r = 0.3384, p = 0.0021) and S. mutans-S. sobrinus (r = 0.3978, p = 0.0003). ANCOVA revealed that dmfs index had a significant effect on the distribution of CFU of S. mutans (p = 0.0118) and S. sobrinus (p = 0.03). When MSB was compared with PCR to identify MS, there was no statistical difference (p > 0.05). We conclude that S. mutans and S. sobrinus were isolated in higher numbers from CA children and those harbouring both bacteria had higher dmfs scores. PCR is a useful tool in molecular epidemiology for dental caries studies; it was effective in detecting and identifying MS from saliva in children. PMID- 18389678 TI - Microtensile bond strength of contemporary adhesives to primary enamel and dentin. AB - The purpose of this study was to assess bond strength of three self-etching and two total-etch adhesive systems bonded to primary tooth enamel and dentin. MATERIALS AND METHODS: Forty extracted primary human molars were selected and abraded in order to create flat buccal enamel and occlusal dentin surfaces. Teeth were assigned to one of the adhesive systems: Adper Scotch Bond Multi Purpose, Adper Single Bond 2, Adper Prompt L-Pop, Clearfil SE Bond and AdheSE. Immediately to adhesive application, a composite resin (Filtek Z250) block was built up. After 3 months of water storage, each sample was sequentially sectioned in order to obtain sticks with a square cross-sectional area of about 0.72 mm2. The specimens were fixed lengthways to a microtensile device and tested using a universal testing machine with a 50-N load cell at a crosshead speed of 0.5 mm/min. Microtensile bond strength values were recorded in MPa and compared by Analysis of Variance and the post hoc Tukey test (a = 0.05). RESULTS: In enamel, Clearfil SE Bond presented the highest values, followed by Adper Single Bond 2, AdheSE and Adper Scotch Bond Multi Purpose, without significant difference. The highest values in dentin were obtained with Adper Scotch Bond Multi Purpose and all other adhesives did not present significant different values from that, except Adper Prompt L-Pop that achieved the lowest bond strength in both substrates. Adper Scotch Bond Multi Purpose and Adper Single Bond 2 presented significantly lower values in enamel than in dentin although all other adhesives presented similar results in both substrates. CONCLUSIONS: contemporary adhesive systems present similar behaviors when bonded to primary teeth, with the exception of the one-step self-etching system; and self-etching systems can achieve bond strength values as good in enamel as in dentin of primary teeth. PMID- 18389679 TI - Pediatricians' perceptions of the use of sweetened medications related to oral health. AB - OBJECTIVE: To evaluate pediatricians' perceptions and attitudes about the use of liquid pediatric medicines and their relationship with dental caries and dental erosion. STUDY DESIGN: A cross sectional study was conducted. Data was collected by questionnaires handed out in hospitals, medical clinics and offices. A convenience sample of 104 pediatricians was obtained. RESULTS: Most respondents (80.8%) stated that pediatric medicines could be related with dental disorders. Dental caries (64.70%) and tooth discoloration (43.7%) were the most frequent mentioned alterations, while only 3 (4.2%) respondents pointed out dental erosion. A considerable number of respondents (62.50%) recognized the presence of fermentable carbohydrates as a contributing factor to tooth decay, however not all of them recommended oral hygiene after their consumption (50.80%). Besides, 48 respondents (46.20) also believed that pediatric medicines could cause dental wear. CONCLUSIONS: Pediatricians in this study did not perceive the correct relationship between the presence of acidity in medicines and dental erosion; however, most of them presented a reasonable awareness about the relationship between sugared pediatric medicines and dental caries. Besides, they were unaware about the need of recommending oral hygiene after medicines' use. PMID- 18389680 TI - A catalogue of anomalies and traits of the primary dentition of southern Chinese. AB - Variation in size, form and morphology of the teeth result in anomalies such as macrodontia, microdontia, hyperdontia, hypodontia, double tooth, taurodontism and dens in dente. While traits that may occur more commonly in certain ethnic groups may be considered to be specific to that population. The characteristics of these anomalies and traits are presented along with the prevalence figures for their occurrence in primary dentition of southern Chinese. PMID- 18389681 TI - Management of Blandin-Nuhn mucocele--a case report. AB - Mucoceles of the glands of Blandin and Nuhn are uncommon and mainly seen on the ventral surface of the tongie. This case report emphasizes the treatment approach of these mucoceles. This patient reported with a second recurrence following conventional treatment, so a novel method was tried by injecting an ultraflow rubber base impression material into the mucocele, after which a surgical excision was done. The present procedure showed a clear demarcated limit of the lesion making the surgical excision easier PMID- 18389682 TI - Oral surgery in a child with a prosthetic aortic valve and pulmonary artery stent at risk for thromboembolism. AB - Children with prosthetic cardiac valves and other invasive mechanical devices that direct blood flow require anticoagulant medication for prevention of thrombosis. Dental surgery for these children has historically consisted of decreasing and/or discontinuing the oral anticoagulant and instituting heparin therapy prior to the planned dental procedure, which can result in thromboembolism and increased morbidity and mortality. This case report demonstrates that oral anticoagulation need not be decreased or discontinued prior to extraction of multiple carious primary teeth in a child at risk for thromboembolism. PMID- 18389683 TI - Caries experience of leukemic children during intensive course of chemotherapy. AB - OBJECTIVE: To determine the incidence of dental caries in 64 leukemic children who received dental treatment before the initiation of chemotherapy and had intensive oral care during the course. DESIGN: Caries experience (deft/DMFT), white-spot lesions (enamel demineralization), premature primary molar extractions and carious first permanent molars were investigated before and after intensive therapy. SETTING: Pediatric Dental Care Unit and Department of Pediatric Hematology, Faculty of Medicine, Uludag University. RESULTS: The dental caries level was insignificantly higher following chemotherapy compared to the initial caries level. However the prevalence of white-spot lesions was significantly increased during the course. Caries level in 30 subjects (47%) remained stable. CONCLUSION: Caries level in leukemic children, who were not caries-free before chemotherapy, could be stabilized by caries prevention methods such as frequent topical fluoride applications and fissure sealants, intensive oral care and improved self-care practices. PMID- 18389684 TI - Assessment of head tilt in young children with unilateral posterior crossbite by video recording. AB - 14 children with unilateral posterior crossbites (PCB) participated in this study and 14 children with Angle's class I occlusion. Body posture analysis was made by a video recording technique. The results showed greater tilt in the angles of head tilt in PCB children when compared to neutral occlusion children. We also observed that head tilt followed the side of crossbite. Such results suggest that unilateral PCB could be related to the development of head tilt on the same side of the crossbite. PMID- 18389685 TI - The quad-helix appliance in the primary dentition--orthodontic and orthopedic measurements. AB - Changes in the midpalatal suture and the alterations in intercanine and intermolar widths with the use of the quad-helix appliance were evaluated. Study casts and occlusal radiographs from 10 patients with a mean age of 4 years and 10 months (SD 11 months) were analyzed. A statistically significant palatal suture widening was observed in all cases. The proportion of dental tipping accomplished with the treatment was greater than opening of the suture. PMID- 18389686 TI - Risk factors for caries in children from south-western Nigeria. AB - OBJECTIVE: The objective of this study was to understand the relationship between relevant socio-economic, biological and behavioral indicators and dental caries experience of Nigerian children. METHOD: A cross sectional study design was utilized. The study population consisted of children who visited two Child Dental Health Clinics in South Western Nigeria. Information collected include demographic data, feeding practices, dietary habit, use of medications and daily frequency of tooth brushing. The number of decayed, filled and missing teeth (dmft/DMFT) was noted for children with caries. A stepwise logistic regression analysis was carried to select the best predictor for the presence of caries and for high caries levels (a dmft/DMFT greater than 3). RESULTS: The only predictor for caries was age with the possibility of having caries being 0.89 (11% reduced odds) for every year increase in age (95% CI 0.82 to 0.97) p = 0.008. However, the predictor for a high caries level was age and gender; the odds of having a dmft/DMFT equal to or higher than 3 was 0.80 (20% reduced odds) for every year increase in age (95% CI 0.72 to 0.88) p < 0.001 and 0.57 (43% reduced odds) for every female child (95% CI 0.33 to 0.98) p= 0.04 compared to a male child. CONCLUSION: Children from this study population who consumed sugar once a day are at risk of developing caries. Also, male children whose daily consumption of sugar was equal to or greater than three times daily, were at risk for having a dmft/DMFT higher than 3. PMID- 18389687 TI - Growth and toxin production of proteolytic Clostridium botulinum in aseptically steamed rice products at pH 4.6 to 6.8, packed under modified atmosphere, using a deoxidant pack. AB - Demand for aseptically steamed rice products has been increasing rapidly in Japan over the past 10 years. In our previous study, we showed that proteolytic Clostridium botulinum produce toxins in steamed rice products packaged under a modified atmosphere of < or =0.3% oxygen. In the present study, we examined the effect of pH to control botulism risk in steamed rice products packaged under modified atmosphere (5% CO2 and 95% N2 as the balance) with the inclusion of a deoxidant pack to produce an oxygen concentration of < or =0.3%. A mixture of 10 strains of proteolytic C. botulinum (5 type A strains and 5 type B strains) was inoculated into steamed rice products at pH values between 4.6 and 6.8 prior to packaging. All samples were stored at 30 degrees C for 24 weeks. Samples at higher pH showed earlier starts of neurotoxin production. Neurotoxin was detected after 2 weeks of incubation in samples at pH 5.4 or above, whereas it took 4 weeks for the toxin to be detected in samples at pH 5.2 to 5.3 and 12 weeks in samples at pH 5.0 to 5.1. In samples at pH 4.9 or below, no toxin was detected during the experimental period. Apparent sample spoilage did not occur before C. botulinum produced neurotoxin in most of the samples. Based on these results, we conclude that aseptically steamed rice products must be packaged at pH 4.9 or below under modified atmosphere containing < or =0.3% oxygen, with the inclusion of a deoxidant pack. PMID- 18389688 TI - Decontamination of fluid milk containing Bacillus spores using commercial household products. AB - Although commercial sanitizers can inactivate bacterial spores in food processing environments, relatively little data exist as to the decontamination of products and surfaces by consumers using commercial household products. Should a large scale bioterrorism incident occur in which consumer food products were contaminated with a pathogenic sporeformer such as Bacillus anthracis, there may be a need to decontaminate these products before disposal as liquid or solid waste. Studies were conducted to test the efficacy of commercial household products for inactivating spores of Bacillus cereus (used as a surrogate for B. anthracis) in vitro and in fluid milk. Validation of the resistance of the B. cereus spores was confirmed with B. anthracis spores. Fifteen commercial products, designed as either disinfectants or sanitizers or as potential sanitizers, were purchased from retail markets. Products selected had one of the following active compounds: NaOCl, HCl, H2O2, acetic acid, quaternary ammonium compounds, ammonium hydroxide, citric acid, isopropanol, NaOH, or pine oil. Compounds were diluted in water (in vitro) or in 2% fat fluid milk, and spores were exposed for up to 6 h. Products containing hypochlorite were most effective against B. cereus spores. Products containing HCl or H2O2 also reduced significant numbers of spores but at a slower rate. The resistance of spores of surrogate B. cereus strains to chlorine-containing compounds was similar to that of B. anthracis spores. Therefore, several household products on the market may be used to decontaminate fluid milk or similar food products contaminated by spores of B. anthracis. PMID- 18389689 TI - Economics of reducing Campylobacter at different levels within the Belgian poultry meat chain. AB - Campylobacter infections pose a serious public health problem in Belgium. Poultry meat is most likely responsible for 40% of human campylobacteriosis cases in Belgium. On a yearly basis, consumption of poultry meat causes at least 22,000 campylobacteriosis cases, with a cost of illness of Euro 10.9 million. Several intervention measures have been proposed in literature, aiming to reduce the contamination of poultry meat and thus lead to significant reductions of human campylobacteriosis cases. This study aimed to evaluate the cost-benefit ratio, i.e., the ratio of reduced costs of illness on intervention costs of various intervention measures. These measures were selected by representatives from the poultry meat sector and experts in the field of poultry science. The selection comprised measures at the farm level (phage therapy), at the processing plant (spraying of carcasses with lactic acid or electrolyzed oxidizing water, crust freezing, or irradiation), and at the consumer level (improving kitchen hygiene and application of home freezing). Among these measures, the decontamination of carcasses with electrolyzed oxidizing water applied in the processing plant was the most efficient (17.66), followed by the use of lactic acid (4.06). In addition, phage therapy generated a positive cost-benefit ratio (2.54). Irradiation indicated the highest efficacy, but its cost-benefit ratio was rather low (0.31). There seems to be less gain by trying to improve food handling in the kitchen. The cost to reach consumers is large, while only a very limited fraction of the consumers is willing to change its behavior. The outcome of this study poses valuable information for future risk-management decisions in Belgium. PMID- 18389690 TI - Inhibition of Escherichia coli O157:H7 in ripening dry fermented sausage by ground yellow mustard. AB - Compounds generated by the enzymatic hydrolysis of glucosinolates naturally present in mustard powder are potently bactericidal against Escherichia coli O157:H7. Because E. coli O157:H7 can survive the dry fermented sausage manufacturing process, 2, 4, and 6% (wt/wt) nondeheated (hot) mustard powder or 6% (wt/wt) deheated (cold) mustard powder were added to dry sausage batter inoculated with E. coli O157:H7 at about 7 log CFU/g to evaluate the antimicrobial effectiveness of the powders. Reductions in E. coli O157:H7 populations, changes in pH and water activity (aw), effects on starter culture (Pediococcus pentosaceus and Staphylococcus carnosus) populations, and effects of mustard powder on sausage texture (shear) were monitored during ripening. Nondeheated mustard powder at 2, 4, and 6% in dry sausage (0.90 aw) resulted in significant reductions in E. coli O157:H7 (P < 0.05) of 3.4, 4.4, and 6.9 log CFU/g, respectively, within 30 days of drying. During fermentation and drying, mustard powder did not affect P. pentosaceus and S. carnosus activity in any of the treatments. Extension of drying to 36 and 48 days reduced E. coli O157:H7 by >5 log CFU/g in the 4 and 2% mustard powder treatments, respectively. The 6% deheated mustard powder treatment provided the most rapid reductions of E. coli O157:H7 (yielding <0.20 log CFU/g after 24 days) by an unknown mechanism and was the least detrimental (P < 0.05) to sausage texture. PMID- 18389691 TI - Effect of inoculum preparation procedure and storage time and temperature on the fate of Listeria monocytogenes on inoculated salami. AB - Although dry/semidry fermented sausages are characterized as being of low-to moderate risk for human listeriosis on a per-serving and per-annum basis, data are lacking relative to the fate of postprocessing Listeria monocytogenes contamination during storage of such products. This study evaluated the effect of inoculum preparation and storage conditions on the fate of L. monocytogenes on vacuum-packaged salami. Commercially produced salami was sliced and inoculated (4 +/- 1.3 log CFU/ cm2) with one of four types of inocula. All inocula consisted of the same 10-strain L. monocytogenes composite, cultivated as individual strains prior to mixing for inoculation. Active cultures of individual strains were prepared (30 degrees C, 24 h) in either tryptic soy broth (containing 0.25% glucose) plus 0.6% yeast extract (TSBYE), tryptic soy broth without glucose plus 0.6% yeast extract (TSBYE-G), TSBYE-G plus 1% glucose (TSBYE+G), or in TSBYE, and then habituated (7 degrees C, 72 h) in sterile salami homogenate (10% [wt/wt] with distilled water). Inoculated salami slices were vacuum packaged, stored at 4, 12, or 25 degrees C, and analyzed (three samples per treatment in each of two replicates) periodically for surviving bacterial counts. In general, pathogen levels decreased during storage and reached levels below the detection limit ( 0.4 log CFU/cm2) between 27 and 90 days of storage, depending on temperature of storage and inoculum type. Death rates (log CFU/cm2/day) were found to increase as storage temperature increased, with the exception of the acid-adapted (TSBYE+G) cells, which decreased more rapidly at 4 degrees C than at 12 or 25 degrees C. The habituated inoculum was inactivated at a faster rate than other inocula at 12 and 25 degrees C, but performed similarly to nonadapted (TSBYE-G) and partially acid-adapted (TSBYE) inocula at 4 degrees C. These data may be used to supplement existing information for use in future risk assessments. PMID- 18389692 TI - Antibacterial efficiency of Spanish Satureja montana essential oil against Listeria monocytogenes among natural flora in minced pork. AB - The purpose of this study was to examine the effectiveness of winter savory (Satureja montana) essential oil (EO) for control of growth and survival of experimentally inoculated Listeria monocytogenes serovar 4b (10(4) CFU/g) among natural flora in minced pork. EOs of French thyme (Thymus vulgaris F) and rosemary (Rosmarinus officinalis) cultivated in the same region of Aragon (northeastern Spain) were used as reference ingredients. The EOs obtained by hydrodistillation were added at concentrations of 0.25, 0.5, 1, and 2.5 microl/g (vol/wt), and the samples were kept at 4 degrees C in air for up to 7 days. The populations of L. monocytogenes and total viable bacteria were determined in the control and treated samples at 0, 1, 3, 5, and 7 days. Moderate activity of S. montana EO against L. monocytogenes was observed (at 2.5 microl/g, reductions of 0.27 log CFU/g by day 3 and 0.61 log CFU/g by day 7), with higher activity against aerobic flora. The greatest reduction in aerobic flora was on day 3 (at 2.5 microl/g) from 1.10 to 1.45 log CFU/g. S. montana EO was comparable to T. vulgaris F EO in listericidal activity, but R. officinalis EO was ineffective against the L. monocytogenes and aerobic flora in the minced meat model. The approximately 3-log reduction in aerobic flora with T. vulgaris F EO at 0.25 to 2.5 microl/g after 5 days of storage was the most significant reduction. Depending on sensory considerations, the addition of active EOs in combination with other preservation techniques for synergistic effects may provide alternatives to synthetic chemical preservatives. Suggestions on relationships between chemical composition and biological activities of EOs are outlined. PMID- 18389693 TI - Thermal inactivation D- and z-values of multidrug-resistant and non-multidrug resistant Salmonella serotypes and survival in ground beef exposed to consumer style cooking. AB - There has been speculation that multidrug-resistant (MDR) strains are generated by subtherapeutic antibiotic use in food animals and that such strains result in increased resistance to lethality by food processes such as heat and irradiation. The objective of this study was to evaluate the heat resistance of 20 strains, namely an MDR and a non-multidrug-resistant (NMDR) strain of each of 10 Salmonella serotypes isolated from cattle or cattle environments. MDR and NMDR Salmonella serotypes studied included Montevideo, Typhimurium, Anatum, Muenster, Newport, Mbandaka, Dublin, Reading, Agona, and Give. For phase I, stationary phase cultures of the strains were aliquoted into sterile capillary tubes and immersed in a temperature-controlled water bath at 55, 60, 65, and 70 degrees C for appropriate times. Survivor curves were plotted for each temperature, and a best-fit linear regression was derived for each temperature. D-values (decimal reduction times) and z-values (changes in temperature required to change the D values) were calculated for each strain. Although there was no overall significant difference in the heat resistance of MDR and NMDR serotypes, NMDR serotypes generally appeared to have slightly higher heat resistance than NMDR serotypes, especially at 55 and 60 degrees C. The highest relative heat resistance (highest z-values) was exhibited by Salmonella Anatum. Notably, the relative heat resistance of NMDR Salmonella Agona was similar to that of NMDR Salmonella Anatum and had the highest D-values at all four temperatures. For phase II, three serotypes (regardless of resistance profile) with the highest relative heat resistance and their drug-resistant counterparts were selected for thermal inactivation in ground beef patties cooked to endpoint temperatures. Salmonella Agona was able to survive in ground beef cooked to an internal temperature of 71 degrees C. Results of these studies suggest drug resistance does not affect the heat resistance of Salmonella and that serotype or strain is an important consideration in risk assessment of the pathogen with regard to survival at cooking temperatures. PMID- 18389694 TI - Effect of selected plant essential oils or their constituents and modified atmosphere packaging on the radiosensitivity of Escherichia coli O157:H7 and Salmonella typhi in ground beef. AB - Twenty-six different essential oils were tested for their efficiency to increase the relative radiosensitivity of Escherichia coli and Salmonella Typhi in medium fat ground beef (23% fat). Ground beef was inoculated with E. coli O157:H7 or Salmonella (10(6) CFU/g), and each essential oil or one of their main constituents was added separately at a concentration of 0.5% (wt/wt). Meat samples (10 g) were packed under air or under modified atmosphere and irradiated at doses from 0 to 1 kGy for the determination of the D10-value of E. coli O157:H7, and from 0 to 1.75 kGy for the determination of the D10-value of Salmonella Typhi. Depending on the compound tested, the relative radiation sensitivity increased from 1 to 3.57 for E. coli O157:H7 and from 1 to 3.26 for Salmonella Typhi. Addition of essential oils or their constituents before irradiation also reduced the irradiation dose needed to eliminate both pathogens. In the presence of Chinese cinnamon or Spanish oregano essential oils, the minimum doses required to eliminate the bacteria were reduced from 1.2 to 0.35 and from 1.4 to 0.5 for E. coli O157:H7 and Salmonella Typhi, respectively. Cinnamon, oregano, and mustard essential oils were the most effective radiosensitizers. PMID- 18389695 TI - Electrostatic forces control nonspecific virus attachment to lettuce. AB - Enteric viruses are key foodborne pathogens. The objective of this study was to compare the relative contributions of electrostatic and hydrophobic forces with the nonspecific attachment of virus to butterhead lettuce. The attachment of four viruses (echovirus 11, feline calicivirus [FCV], MS2, and phiX174) was studied. Three different conditions, namely (i) 1% Tween 80, (ii) 1 M NaCl, and (iii) 1% Tween 80 with 1 M NaCl, were investigated to determine the role of hydrophobic, electrostatic, and combined hydrophobic and electrostatic forces, respectively. Attachment above the pI of FCV and echovirus 11 was reduced or eliminated in the presence of NaCl, indicating an electrostatic interaction between the animal viruses and lettuce. The bacteriophage phiX174 was not significantly affected by any treatment, indicating a lack of electrostatic or hydrophobic interactions between the lettuce and phage phiX174. Overall, 1 M NaCl was the most effective treatment in desorbing viruses from the surface of lettuce at pH 7 and 8. The results imply that electrostatic forces play a major role in controlling virus adsorption to lettuce. The results indicate that 1 M NaCl solution would improve the recovery or elution of unenveloped viruses from lettuce. PMID- 18389696 TI - Potential sources of microbial contamination of satsuma mandarin fruit in Japan, from production through packing shed. AB - Potential sources of microbial contamination of satsuma mandarin fruit were investigated from production through the packing shed in the 2005 season. Microbial counts in the peel and flesh during the fruit development stage were below 2.4 log CFU/g for bacteria and 3 log CFU/g for fungi, except for the peel in August and September. In the field environment, the highest microbial counts were found in fallen leaves on the ground, followed by soil, organic fertilizer, and agricultural water. Only the pesticide solution collected in July was positive for Salmonella, while no verotoxin-producing Escherichia coli was detected from any of the samples. The bacterial and mold flora in the peel comprised phytopathogenic organisms such as bacteria genus Pantoea and mold genus Mycosphaerella and soilborne organisms such as bacteria genus Bacillus and mold genus Cladosporium, which were found in soil, fallen leaves, agricultural water, and cloth mulch throughout the production season. After fruit harvest and sorting, microbial counts of the peel increased, while those of the flesh remained below the lower limit of detection. Although some of the preharvest sources could also be postharvest sources, some packing shed equipment was assumed to be postharvest sources, because Bacillus cereus was not identified from the fruit in the production field but was detected on the peel after sorting and on equipment such as gloves, plastic harvest basket, and size sorter. These results suggest that using sanitizers for agricultural water and packing sheds to prevent cross-contamination would be useful in a good agricultural practices program of the satsuma mandarin in Japan. PMID- 18389697 TI - Comparative effect of direct-fed microbials on fecal shedding of Escherichia coli O157:H7 and Salmonella in naturally infected feedlot cattle. AB - The effect of direct-fed microbials (DFM) on fecal shedding of Escherichia coli O157:H7 and Salmonella in naturally infected feedlot cattle was evaluated in a clinical trial involving 138 feedlot steers. Following standard laboratory methods, fecal samples collected from steers were evaluated for change in the detectable levels of E. coli O157:H7 and Salmonella shed in feces after DFM treatment. Sampling of steers was carried out every 3 weeks for 84 days. A significant reduction (32%) in fecal shedding of E. coli O157:H7 (P < 0.001), but not Salmonella (P = 0.24), was observed among the treatment steers compared with the control group during finishing. The probability of recovery of E. coli O157:H7 from the feces of treated and control steers was 34.0 and 66.0%, respectively. Steers placed on DFM supplement were almost three times less likely to shed E. coli O157:H7 (odds ratio, 0.36; 95% confidence interval, 0.25 to 0.53; P < 0.001) in their feces as opposed to their control counterparts. The probability of recovery of Salmonella from the feces of the control (14.0%) and the treated (11.3%) steers was similar. However, the DFM significantly reduced probability of new infections with Salmonella among DFM-treated cattle compared with controls (nontreated ones). It appears that DFM as applied in our study are capable of significantly reducing fecal shedding of E. coli O157:H7 in naturally infected cattle but not Salmonella. The factors responsible for the observed difference in the effects of DFM on E. coli O157:H7 and Salmonella warrants further investigation. PMID- 18389698 TI - Low prevalence of Listeria monocytogenes in cull sows and pork. AB - The goal of this study was to determine the prevalence of Listeria monocytogenes in sows slaughtered at a single Midwestern plant on two occasions (trial 1, n = 179 sows; trial 2, n = 160 sows). Fecal samples collected antemortem (trial 1) as well as animal tissues, and carcass swabs collected at the abattoir (trials 1 and 2) were analyzed. Eight isolates of L. monocytogenes were recovered from five samples that represented 0.18% of the total samples (n = 2,775). In trial 1, L. monocytogenes was detected in a tonsil sample (0.6%; 1 positive of 181 tonsils), in a carcass (0.6%; 1 positive of 179 carcasses), which was sampled prior to the organic rinse, and in two chopped meat block samples (1.2%; 2 positive of 165 samples). In trial 2, L. monocytogenes was only detected in a single chopped meat block sample (0.15%; 1 positive of 688 total samples). These data indicate the low prevalence of L. monocytogenes in the cull sow. PMID- 18389699 TI - Are staff management practices and inspection risk ratings associated with foodborne disease outbreaks in the catering industry in England and Wales? AB - Despite structured enforcement of food hygiene requirements known to prevent foodborne disease outbreaks, catering businesses continue to be the most common setting for outbreaks in the United Kingdom. In a matched case control study of catering businesses, 148 businesses associated with outbreaks were compared with 148 control businesses. Hazard analysis critical control point systems and/or formal food hygiene training qualifications were not protective. Food hygiene inspection scores were not useful in predicting which catering businesses were associated with outbreaks. Businesses associated with outbreaks were more likely to be larger small and medium-sized enterprises (SMEs) or to serve Chinese cuisine and less likely to have the owner or manager working in the kitchen, but when size of the SME was taken into account these two differences were no longer significant. In larger businesses, case businesses were more likely to be hotels and were more commonly associated with viral foodborne outbreaks, but there was no explanation within the data for this association. PMID- 18389700 TI - Complete validation of a unique digestion assay to detect Trichinella larvae in horse meat demonstrates the reliability of this assay for meeting food safety and trade requirements. AB - A tissue digestion assay using a double separatory funnel procedure for the detection of Trichinella larvae in horse meat was validated for application in food safety programs and trade. The assay consisted of a pepsin-HCl digestion step to release larvae from muscle tissue and two sequential sedimentation steps in separatory funnels to recover and concentrate larvae for detection with a stereomicroscope. With defined critical control points, the assay was conducted within a quality assurance system compliant with International Organization for Standardization-International Electrotechnical Commission (ISO/IEC) 17025 guidelines. Samples used in the validation were obtained from horses experimentally infected with Trichinella spiralis to obtain a range of muscle larvae densities. One-, 5-, and 10-g samples of infected tissue were combined with 99, 95, and 90 g, respectively, of known negative horse tissue to create a 100-g sample for testing. Samples of 5 and 10 g were more likely to be positive than were 1-g samples when larval densities were less than three larvae per gram (lpg). This difference is important because ingested meat with 1 lpg is considered the threshold for clinical disease in humans. Using a 5-g sample size, all samples containing 1.3 to 2 lpg were detected, and 60 to 100% of samples with infected horse meat containing 0.1 to 0.7 lpg were detected. In this study, the double separatory funnel digestion assay was efficient and reliable for its intended use in food safety and trade. This procedure is the only digestion assay for Trichinella in horse meat that has been validated as consistent and effective at critical levels of sensitivity. PMID- 18389701 TI - Real-time PCR for quantitative detection of bovine tissues in food and feed. AB - A real-time PCR approach with the SYBR Green detection system has been developed for the quantitative detection of bovine tissues in food and feedstuffs. The method combines the use of bovine-specific primers, which amplify an 84-bp fragment of the mitochondrial 12S rRNA gene, and universal primers, which amplify a 140-bp fragment of the nuclear 18S rRNA gene from eukaryotic DNA. The 18S rRNA primers are used as endogenous controls for the total content of PCR-amplifiable DNA in the sample. The specificity of the primers was tested against 18 animal species, including mammals, birds, and fish, as well as 6 plant species. Analysis of experimental bovine tissues-oats mixtures demonstrated the suitability of the assay for the detection of bovine DNA in mixtures containing as low as 0.1% of bovine tissues. The performance of the method is not affected by severe heat treatment (up to 133 degrees C for 20 min at 300 kPa). The reported PCR assay could be very useful for detecting bovine-derived ingredients in raw and heat treated food and feedstuffs. PMID- 18389702 TI - Estimated compliance for removal of specified risk materials from 18 U.S. beef packing plants. AB - The removal of 18,345 specified risk materials was observed during audits of 18 U.S. beef processing facilities that, in total, account for over 90% of total U.S. beef slaughtered. Audited plants varied in capacity (280 to 6,000 head per day) and processed both "fed (young cattle)" and "nonfed (mature cows/bulls)" cattle. When all observations for removal of specified risk materials were combined from plants and adjusted for type of cattle processed, overall compliance with specified risk material removal regulations was 98.08%. A 100% compliance rate for removal of brains and distal ileums was recorded based on a total of 600 observations for removal of brains and a total of 2,400 observations for removal of distal ileums. Observations for removal of dorsal root ganglia were collected from 16 of the 18 plants, and overall compliance for dorsal root ganglia removal was 99.6% (4,783 of 4,800). Fifteen of the 16 plants were 100% compliant. For tonsils, data from 18 plants were collected, and tonsils were correctly removed from 92.8% (4,777 of 5,145) of tongues and heads. Data for spinal cord removal were collected from 18 plants, and the spinal cord was removed completely in line with U.S. Department of Agriculture-Food Safety and Inspection Service regulations for 99.43% of the observations. Based on the results of this study, packing plants have demonstrated that they are complying with regulations for removal of specified risk materials from beef meat products intended for human consumption greater than 98% of the time. To continue to assure food safety and consumer confidence, continued vigilance and provision of training programs for plant workers are essential. PMID- 18389703 TI - European interlaboratory trial regarding the official microscopic method for the detection of the presence of animal constituents in feedstuffs. AB - The bovine spongiform encephalopathy epidemic is thought to have occurred as a consequence of feeding prion-infected material to cattle. To avoid the risk of bovine spongiform encephalopathy diffusion, the European Commission (Directive 2003/126/EC) established an official method to detect the presence of animal derived constituents in feedstuffs, using microscopic examination. This method allows easy identification of bone fragments among other animal constituents. The analysis is based on morphological conformation of the fragments and their characterization (mainly of the shape of lacunae) to discriminate among mammalian, poultry, and fish tissues. The aim of this study was to assess the performances of nine European laboratories through a ring trial of the official microscopic method, and to calculate accuracy and reproducibility of the method. In general the reproducibility of the microscopic method was very good (kappa overall = 0.83), with a high sensitivity for all laboratories. Concerning the analysis on the different animal-derived constituents, the results show values of sensitivity with large variability between fish and poultry or mammal. It was generally more difficult to discriminate between mammalian and poultry tissues than fish tissue. PMID- 18389704 TI - Sulfamethazine water medication pharmacokinetics and contamination in a commercial pig production unit. AB - Sulfamethazine is often used to treat disease in the swine industry. Sulfamethazine is available as water or feed medication and historically (over the past 40 years) has been associated with residue violations in both the United States and Europe. Despite sulfamethazine's approval for use as a water medication, little research on the pharmacokinetics of the water formulation is available. Therefore, a pilot study was performed to determine the plasma levels of an approved sulfamethazine water medication. Plasma levels in pigs treated with an oral bolus (250 mg/kg), which is equivalent to the total drug consumed within a 24-h period, achieved therapeutic concentrations (50 microg/ml). Noncompartmental-based pharmacokinetic model parameters for clearance, half-life, and volume of distribution were consistent with previously published values in swine. However, the above treatment resulted in exposure of pen mates to sulfamethazine at levels currently above tolerance (0.1 ppm). Using a physiologically based pharmacokinetic model, the treatment dose simulation was compared with observed plasma levels of treated pigs. Flexibility of the physiologically based pharmacokinetic model also allowed simulation of control pig plasma levels to estimate contamination exposure. A simulated exposure to 0.15 mg/kg twice within approximately 8 h resulted in detectable levels of sulfamethazine in the control pigs. After initial exposure, a much lower dose of 0.059 mg/kg maintained the contamination levels above tolerance for at least 3 days. These results are of concern for producers and veterinarians, because in commercial farms, the entire barn is often treated,and environmental contamination could result in residues of an unknown duration. PMID- 18389705 TI - Detection of melamine using commercial enzyme-linked immunosorbent assay technology. AB - Recent cases of adulteration with melamine have led to the need for rapid and reliable screening methods. To meet this need, commercial enzyme-linked immunosorbent assay (ELISA) test kits for the detection of triazines were evaluated. The recently released Melamine Plate kit (Abraxis, Warminster, Pa.) displayed a limit of detection of 9 ng/ml for melamine in phosphate-buffered saline (PBS) and approximately 1 microg/ml for melamine added to dog food. An atrazine ELISA test kit produced by Abraxis required 0.2 mg/ml to generate a response more than four times the standard deviation from background. In contrast, with the EnviroGard Triazine Plate kit (Strategic Diagnostics, Inc., Newark, Del.), 1.5 mg/ml melamine in PBS generated a signal only one standard deviation from background, which was insufficient to define a limit of detection. Extraction based on dilution with 105 mM sodium phosphate/75 mM NaCl/2.5% nonfat milk/0.05% Tween 20 (UD) enabled detection of fivefold less melamine in dog food than did use of the procedure recommended by the manufacturer, which entailed extraction into 60% methanol, sonication, centrifugation, filtration, and further dilution into 10% methanol/PBS. Using the Abraxis Melamine ELISA, both extraction protocols yielded identical results with a dog food sample adulterated with melamine. The recovery of melamine spiked into gravy from dog food using UD was 74% +/- 4%. In conclusion, the recently released Abraxis ELISA for melamine proved to be a useful alternative to more cumbersome methods. PMID- 18389706 TI - Trial for quality control in mercury contents by using tail muscle of full-cycle cultured bluefin tuna (Thunnus orientalis). AB - Substantial amounts of mercury are usually present in tuna muscle, with levels in excess of 10 times the standard safety value present in some individuals. Inspection of individual fish for mercury content would be desirable but may not be cost-effective. In this study, we tried to establish a low-cost system for checking the mercury content of tuna by using a tail muscle that is usually discarded. The samples used in this experiment were bluefin tuna, cultured in the Fisheries Laboratory of Kinki University (Oshima Experimental Station, Wakayama, Japan). They were raised from eggs spawned in 2002. Ninety-eight individuals, weighing 22.3 to 61.6 kg, were selected between December 2004 and November 2005. In nine individuals, the mercury content of the tail was compared with that of the whole body. The total mercury level was measured using the reduction vaporizing atomic absorption method after acid digestion. Except for the front of the abdomen, where the mercury content was lower (0.490 ppm), the mercury content of other parts of the fish did not differ from that of the tail muscle (0.631 ppm). Therefore, the overall mercury concentration in bluefin tuna could be estimated to be almost the same and/or lower than that of the tail muscle. On the basis of these results, for 1 year we investigated the quantity of mercury in full-cycle cultured bluefin tuna that were shipped. The mercury concentration showed no increase irrespective of increases of body weight. PMID- 18389707 TI - Comparison of antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from humans and chicken carcasses in Poland. AB - Campylobacter-associated gastroenteritis remains an important cause of morbidity worldwide, and some evidence suggests that poultry is an important source of this foodborne infection in humans. This study was conducted to analyze the prevalence and genetic background of resistance of 149 Campylobacter jejuni and 54 Campylobacter coli strains isolated from broiler chicken carcasses and from stool samples of infected children in Poland from 2003 through 2005. Nearly all isolates were susceptible to macrolides and aminoglycosides. The highest resistance in both human and chicken strains was observed for ciprofloxacin (more than 40%), followed by ampicillin (13 to 21%), and tetracycline (8 to 29%). Resistance to ampicillin and tetracycline rose significantly between 2003 and 2005. Slight differences in resistance between human and chicken isolates indicate that although chicken meat is not the only source of Campylobacter infection in our population, it can be involved in the transmission of drug resistant Campylobacter strains to humans. PMID- 18389708 TI - Occurrence of Listeria and Enterobacteriaceae in domestic refrigerators. AB - Consumers' refrigeration practices have a significant impact on the safety and quality of foods. To determine the prevalence and the identity of microorganisms in domestic refrigerators, swab samples were taken from various locations in the refrigerators from 137 households in middle Tennessee. The swabs were inoculated into different media, and standard procedures were used to characterize the isolates. API 20E and API Listeria were used for identification of Enterobacteriaceae and Listeria spp., respectively. The Kirby-Bauer technique was used to test resistance of the isolates. Actual counts for aerobic and Enterobacteriaceae ranged from not detected to 8.53 and 8.39 log CFU per sample, respectively. Klebsiella pneumoniae (23.4%), Klebsiella oxytoca (6.8%), Klebsiella terrigena (4.0%), Enterobacter sakazakii (2.2%), and Yersinia enterocolitica (0.7%) were some of the bacteria of concern that were isolated from domestic refrigerators. Resistance to antibiotics was most common in erythromycin (39.9%), followed by ampicillin (33.8%), cefoxitin (12.8%), tetracycline (5%), streptomycin (4.0%), nalidixic acid (2.1%), kanamycin (1.4%), and colistin (0.7%). None of the isolates tested was resistant to ciprofloxacin or gentamycin. Listeria spp. were also detected in six refrigerators. These findings underline the need for greater consumer education regarding proper refrigerator cleaning and safe food handling practices. PMID- 18389709 TI - Bacteriological profile of raw, frozen chicken nuggets. AB - The bacteriological profile of raw, frozen chicken nuggets manufactured at a chicken processing facility in Queensland, Australia, was determined. Chicken nuggets are manufactured by grinding poultry, adding premixes to incorporate spices, forming the meat to the desired size and shape, applying a batter and breading, freezing, and packaging. A total of 300 frozen batches were analyzed for aerobic plate count, Escherichia coli, and Salmonella over a period of 4 years. The mean of the aerobic plate count was 5.4 log CFU/g, and counts at the 90th, 95th, and 99th percentiles were 5.7, 5.9, and 6.5 log CFU/g, respectively. The maximum number of bacteria detected was 6.6 log CFU/g. E. coli prevalence was 47%, and of the positive samples, the mean was 1.9 log CFU/g; counts at the 90th, 95th, and 99th percentiles were 2.3, 2.4, and 2.8 log CFU/g, respectively. The maximum number of E. coli was 2.9 log CFU/g. The Salmonella prevalence was 8.7%, and 57.7% of these isolates were typed as Salmonella subspecies II 4,12,[27]:b:[e,n,x] (Sofia), a low-virulence serotype well adapted to Australian poultry flocks. There was a significant relationship (P < 0.05) between season and both aerobic plate counts and E. coli counts, and no correlation between E. coli counts and Salmonella prevalence. This study provides valuable data on the bacteriological quality of raw, frozen chicken nuggets. PMID- 18389710 TI - Microbiological evaluation of stuffed mussels. AB - Stuffed mussel is a traditional food, sold by street vendors in coastal parts of Turkey and other Mediterranean countries. In the present study, the microbiological quality of not only the stuffing mixture, but also the outer surface of the stuffed mussels was evaluated for 1 year, and the effect of the ambient temperatures on the prevalence and the count levels of the microorganisms were evaluated. Fifty samples (750 stuffed mussels in total) were collected periodically, and microbiological analyses were performed by standard procedures for aerobic plate count, coliforms, fecal coliforms, Enterobacteriaceae, Staphylococcus aureus, Bacillus cereus, and Vibrio spp. Aerobic plate counts above 5 log CFU/g were obtained in 16 and 72% of stuffing mixture samples at high and low ambient temperatures, respectively, and average aerobic plate counts of outer surface samples at high and low ambient temperatures were 3.21 and 4.34 log CFU/ml, respectively. The prevalence and the count levels of coliforms, fecal coliforms, Enterobacteriaceae, and Vibrio spp. (except for the prevalence of Vibrio spp. in stuffing mixture samples) in the samples at high ambient temperatures were considerably higher compared with those at low ambient temperatures (P < 0.05). High frequencies of pathogens S. aureus and B. cereus were found in stuffing mixture samples at high ambient temperatures, with averages of 2.84 and 2.94 log CFU/g, respectively (P < 0.05). The result of this investigation indicates that stuffed mussels as a street food may constitute a potential health hazard, especially at high ambient temperatures, depending on contamination level and lack of sanitary practices, and therefore, handling practices should require more attention and improvement. PMID- 18389711 TI - Evaluation of various antimicrobial interventions for the reduction of Escherichia coli O157:H7 on bovine heads during processing. AB - The effectiveness of electrolyzed oxidizing water, FreshFx, hot water, DL-lactic acid, and ozonated water was determined using a model carcass spray-washing cabinet. A total of 140 beef heads obtained from a commercial processing line were inoculated with Escherichia coli O157:H7 on the cheek areas. Each head was exposed to a simulated preevisceration wash and then had antimicrobial wash treatments. Hot water, lactic acid, and FreshFx treatments reduced E. coli O157:H7 on inoculated beef heads by 1.72, 1.52, and 1.06 log CFU/cm2, respectively, relative to the simulated preevisceration wash. Electrolyzed oxidizing water and ozonated water reduced E. coli O157:H7 less than 0.50 log CFU/cm2. Hot water, lactic acid, and FreshFx could be used as decontamination washes for the reduction of E. coli O157:H7 on bovine head and cheek meat. PMID- 18389712 TI - Effect of acidified sodium chlorite, chlorine, and acidic electrolyzed water on Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes inoculated onto leafy greens. AB - Recent foodborne outbreaks implicating spinach and lettuce have increased consumer concerns regarding the safety of fresh produce. While the most common commercial antimicrobial intervention for fresh produce is wash water containing 50 to 200 ppm chlorine, this study compares the effectiveness of acidified sodium chlorite, chlorine, and acidic electrolyzed water for inactivating Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes inoculated onto leafy greens. Fresh mixed greens were left uninoculated or inoculated with approximately 6 log CFU/g of E. coli O157:H7, Salmonella, and L. monocytogenes and treated by immersion for 60 or 90 s in different wash solutions (1:150, wt/vol), including 50 ppm of chlorine solution acidified to pH 6.5, acidic electrolyzed water (pH 2.1 +/- 0.2, oxygen reduction potential of 1,100 mV, 30 to 35 ppm of free chlorine), and acidified sodium chlorite (1,200 ppm, pH 2.5). Samples were neutralized and homogenized. Bacterial survival was determined by standard spread plating on selective media. Each test case (organism x treatment x time) was replicated twice with five samples per replicate. There was no difference (P > or = 0.05) in the time of immersion on the antimicrobial effectiveness of the treatments. Furthermore, there was no difference (P > or = 0.05) in survival of the three organisms regardless of treatment or time. Acidified sodium chlorite, resulted in reductions in populations of 3 to 3.8 log CFU/g and was more effective than chlorinated water (2.1 to 2.8 log CFU/g reduction). These results provide the produce industry with important information to assist in selection of effective antimicrobial strategies. PMID- 18389713 TI - Use of UV light for the inactivation of Listeria monocytogenes and lactic acid bacteria species in recirculated chill brines. AB - Ready-to-eat meat products have been implicated in several foodborne listeriosis outbreaks. Microbial contamination of these products can occur after thermal processing when products are chilled in salt brines. The objective of this study was to evaluate UV radiation on the inactivation of Listeria monocytogenes and lactic acid bacteria in a model brine chiller system. Two concentrations of brine (7.9% [wt/wt] or 13.2% [wt/wt]) were inoculated with a approximately 6.0 log CFU/ml cocktail of L. monocytogenes or lactic acid bacteria and passed through a UV treatment system for 60 min. Three replications of each bacteria-and-brine combination were performed and resulted in at least a 4.5-log reduction in microbial numbers in all treated brines after exposure to UV light. Bacterial populations were significantly reduced after 5 min of exposure to UV light in the model brine chiller compared with the control, which received no UV light exposure (P < 0.05). The maximum rate of inactivation for both microorganisms in treated brines occurred between minutes 1 and 15 of UV exposure. Results indicate that in-line treatment of chill brines with UV light reduces the number of L. monocytogenes and lactic acid bacteria. PMID- 18389714 TI - Influence of peroxyacetic acid and nisin and coculture with Enterococcus faecium on Listeria monocytogenes biofilm formation. AB - Biofilm formation is a matter of concern in food industries because biofilms facilitate the survival of pathogenic bacteria such as Listeria monocytogenes, which may contaminate food-processing equipment and products. In this study, nisin and two Enterococcus faecium strains were evaluated for their effect on biofilm formation by L. monocytogenes cultured in brain heart infusion broth and on stainless steel coupons. Elimination of preformed L. monocytogenes biofilms by peroxyacetic acid also was tested. Adhesion control experiments were performed with pure cultures of L. monocytogenes after swab collection of adhered cells, which were then enumerated on PALCAM agar plates and visualized by scanning electron microscopy. Formation of a biofilm was recorded when the number of adhered cells was at least 10(3) CFU/cm2. When L. monocytogenes was cocultured with E. faecium bac-, the number of adhered L. monocytogenes cells was 2.5 log lower (P = 0.002) when initially compared with the control culture, but after 6 h of incubation a biofilm was again detected. However, in coculture on stainless steel coupons, E. faecium bac+ inhibited L. monocytogenes adherence and did not allow biofilm formation for up to 48 h (P < 0.001). In the presence of nisin or after treatment with peroxyacetic acid, bacterial growth was reduced (P < 0.001) up to 4.6 and 5.6 log CFU/cm2, respectively, when compared with L. monocytogenes cultures on untreated coupons. However, after these treatments, cells were still present, and after 24 h of incubation, a renewed biofilm was detected in L. monocytogenes cultures treated with nisin. Although all tested conditions reduced L. monocytogenes growth to some extent, only coculture with E. faecium bac+ efficiently reduced biofilm formation, suggesting a potential control strategy for this pathogen. PMID- 18389715 TI - Microbial growth and the effects of mild acidification and preservatives in refrigerated sweet potato puree. AB - Refrigerated sweet potato puree is a convenient form of sweet potato that can be used as an ingredient in formulated foods. The microbiology of refrigerated sweet potato puree during storage for up to 5 weeks was evaluated. Because the puree was made by comminuting steam-cooked sweet potatoes before refrigeration, no naturally occurring vegetative bacterial cells were detected during a 4-week period of refrigerated storage at 4 degrees C. However, if postprocessing microbial contamination of the puree were to occur, contaminating microorganisms such as Listeria monocytogenes could grow during refrigerated storage. The effects of acidification or the addition of potassium sorbate and sodium benzoate on a population of L. monocytogenes inoculated into refrigerated (4 degrees C) sweet potato puree were determined. Inoculation of the refrigerated puree with L. monocytogenes at 10(6) CFU/ml resulted in a 3-log increase after 3 weeks storage of nonsupplemented puree. Supplementation of the sweet potato puree with 0.06% (wt/vol) sorbic acid or benzoic acid plus mild acidification of the sweet potato puree with citric acid to pH 4.2 prevented growth of L. monocytogenes during storage at 4 degrees C. PMID- 18389716 TI - Efficacy of sanitizing treatments against Penicillium expansum inoculated on six varieties of apples. AB - The effectiveness of several wash treatments was evaluated against spores of Penicillium expansum inoculated on six varieties of apples (Red Delicious, Golden Supreme, Empire, Macintosh, Fuji, and Gala). The wash treatments were water, acidified water (pH 6.5), acidified sodium hypochlorite (pH 6.5), nonacidified sodium hypochlorite (pH 8.8, 9.3, and 9.7; 50, 100, and 200 ppm, respectively), and peracetic acid (50 and 80 ppm). Spores of P. expansum were dried on the surface of the apples for 2 h before exposure to the different sanitizer solutions. Each apple was submerged in 100 ml of each treatment solution for 30 s, and the number of spores remaining were recovered and enumerated. The efficacy of chlorine solutions was enhanced by decreasing the pH to 6.5 (up to 5-log reduction, depending on apple variety). Peracetic acid solutions (50 and 80 ppm) resulted in a reduction of less than 2 log spores per g and had the same efficacy (P < or = 0.05) as nonacidified chlorine solutions (50, 100, and 200 ppm). Control water solutions produced a reduction of 1.34 log spores per g. Chlorine solutions at pH 6.5 resulted in the largest reduction of P. expansum spores for all apple varieties tested. PMID- 18389717 TI - Development of an experimental model to assess the ability of Escherichia coli O157:H7-inoculated fecal pats to mimic a super shedder within a feedlot environment. AB - This study was conducted to develop an experimental model that could assess the ability of Escherichia coli O157:H7-inoculated fecal pats to mimic a super shedder (>10(4) CFU/g of feces) within a feedlot environment. The day before the study began, 48 steers that had been negative for E. coli O157:H7 in feces for three consecutive weeks were sorted into three treatment groups, with two replicate pens per treatment and 8 steers per pen. Steers within the pens (20.50 by 10.75 m) were exposed to control feces or feces inoculated with two levels of a mixture of five strains of nalidixic acid-resistant E. coli O157:H7 (low level, 10(2) CFU/g; high level, 10(5) CFU/g). Five 300-g fecal pats were introduced into the pens twice daily (10:00 a.m. and 2:30 p.m.) on days 0 through 6 and days 14 through 20. Pats were placed in the pen at random locations to mimic defecation of a steer within the pen. Fecal grab samples, hide swab samples (500-cm2 area of the rump), natural fecal pat samples (freshly voided), and rope samples (1.22-m long manila rope) where obtained at multiple times during the 49-day trial to evaluate the spread of nalidixic acid-resistant E. coli O157:H7 throughout the feedlot environment and among penmates. Immunomagnetic separation and selective media were used to detect E. coli O157:H7. Nalidixic acid-resistant E. coli O157:H7 was detected in 13 high-level treatment fecal grab samples, 7 high-level treatment hide swab samples, 1 low-level hide swab sample, and 2 high-level rope samples. For both fecal grab and hide swab samples, the overall prevalence of E. coli O157:H7 in the high-level group was greater (P < 0.01) than that for the pooled low-level and control groups. Addition of inoculated fecal pats to pens increased transmission of E. coli O157:H7 among penmates, but cattle that acquired E. coli O157:H7 shed the bacterium for only a short time at low levels. Transmission of E. coli O157:H7 from the feces of super shedders to naive penmates may contribute to the observed transient nature of shedding of E. coli O157:H7 among feedlot cattle. PMID- 18389718 TI - Sterility testing of a dispensing valve for aseptic function in food service applications. AB - Manual dispensing equipment for aseptically packaged beverages or foods requires refrigeration of the product package following breakage of the hermetic seal. The food service industry would benefit greatly by implementing dispensing equipment that maintains the sterility of products after continued use without the need for refrigeration. The purpose of this study was to evaluate the ability of a valve, designed to operate aseptically and dispense products with or without refrigeration, to maintain the sterility of products following breakage of the hermetic seal and continued use simulating that of food service. Plastic packages equipped with the "aseptic" dispensing valve in a bag-in-box (BIB) format were aseptically filled with enrichment media with and without the addition of 1% cornstarch to simulate high- and low-viscosity products, respectively. BIBs filled with media were left uninoculated or inoculated (10(4) CFU/ml) with Staphylococcus aureus or Aspergillus niger on the interior of the spout 1 cm from the opening to simulate consumer misuse. Uninoculated and inoculated BIBs were stored at 25 degrees C, and media were dispensed once to twice per day, every day for up to 30 days to simulate food service use. Dispensates were observed for turbidity (compared with controls) indicating growth in BIBs and, thus, breach of sterility. Growth of samples taken aseptically through the package wall was checked microbiologically every 5 days by standard plating techniques. There was no breach in sterility until day 25. At day 25, uninoculated BIB (1 of 45 samples positive for growth) containing high-viscosity media and BIB inoculated with S. aureus (1 of 45 samples positive for growth) containing low-viscosity media became turbid. Viscosity and type of organism did not appear to influence the ability of the valve to maintain the sterility for > or =20 days. The results of this study provide evidence that the dispensing valve tested can maintain the sterility of aseptically filled products following initial dispensation and continued use under unrefrigerated conditions. PMID- 18389719 TI - Characterization of a second ornithine decarboxylase isolated from Morganella morganii. AB - The genes involved in the putrescine formation by Morganella morganii were investigated because putrescine is an indicator of food process deterioration. We report here on the existence of a new gene for ornithine decarboxylase (ODC) in M. morganii. The sequenced 5,311-bp DNA region showed the presence of four complete and one partial open reading frame. Putative functions have been assigned to several gene products by sequence comparison with the proteins included in the databases. The third open reading frame (speC) encoded a 722 amino acid protein showing 70.9% identity to the M. morganii ODC previously characterized (SpeF). The speC gene has been expressed in Escherichia coli, resulting in ODC activity. The presence of a functional promoter (PspeC) located upstream of speC has been demonstrated. Quantitative real-time reverse transcription PCR assay was used to quantify expression of both M. morganii ODC encoding genes, speC and speF, under different growth conditions. This assay allows us to identify SpeF as the inducible M. morganii ODC, since it was highly expressed in the presence of ornithine. PMID- 18389720 TI - Molecular analysis of the first avian influenza H5N1 isolates from fowl in Romania. AB - Since the events of avian influenza (AI) caused by H5N1 subtype from Hong Kong (1997), the people worldwide have been confronted with new waves of epizootic influenza. In 2005 in Romania an unprecedent H5N1 epizootic occurred in domestic and wild birds. Therefore an immediate investigation by molecular approach of this highly pathogenic H5N1 strain was necessary. The virus isolation and the RNA extraction were performed in the Institute of Diagnosis and Animal Health while PCR and sequencing were carried out in Cantacuzino Institute. Herein we report the first evidence of H5N1 presence in Romanian fowls. The phylogenetic analysis of haemagglutinin and neuraminidase gene indicated a close relationship of Romanian strains to those from Siberia and China. The virological and molecular analysis of the first strains of avian virus from Romania confirmed the presence of H5N1 subtype, belonging to the genetic line Z. These results indicate that the avian virus from this genetic line is directly derived from the highly pathogenic viruses isolated in China and Russia in 2005. PMID- 18389721 TI - The viral bronchiolites diagnosis in children by PCR multiplex. AB - The aim of the study was to determine the etiology of the viral bronchiolites in children by using direct immunofluorescence test and 3 RT-PCR Multiplex (S.Bellau Pujol) The study was performed on 122 nasal inspirations collected from 3 weeks-6 month old children hospitalizated in the pediatrics service of CH Rouen. The results were that the majority (53%) of bronchiolites in children had like etiology RSV and a lot of these infections had double viral etiology (26% RSV+ Rhinovirus; 2,7% RSV+HMPV and 1% RSV+Coronavirus 229E). An important viral factor which gives bronchiolitis in children is HMPV (11%). We also find respiratory infections with triple viral etiology: RSV+Influenza A virus + Rhinovirus. PMID- 18389722 TI - Antibodies against HBV, HCV and HAV detected by using microparticle enzyme immunoassay in hepatitis outpatients. AB - The presence of the hepatitis B surface antigen (HBsAg), of the antibodies against HBc, HCV and HAV was determined in outpatients in the period September 2005 - December 2006. The serum samples were analyzed by using Enzyme Immunoassay microparticles (Abbott AxSYM System). At least one test was positive in 238 patients (15.4%) of the total of 1547 patients. Of the 238 positive subjects, in 130 positive subjects (54.6%) the existence of HBV infection could be ascertained based on the presence of HBsAg or of the antibodies against HBc or of their association; 83 patients (34.9%) presented antibodies against HCV and in other 12 patients the antibodies against HCV were associated with HBsAg or with antibodies against HBc, suggesting the coexistence of HCV and HBV infection. The antibodies against HCV and the associations between HCV and HBV were mostly detected in subjects with the diagnosis of cirrhosis, liver failure or chronic hepatitis. Of the 13 (5.46%) patients with antibodies against HAV, 6 patients presented the associations: in 2 cases antibodies anti-HAV with positive HBsAg, in 1 case antibodies anti-HAV and anti-HBc with positive HBsAg, in 2 cases antibodies anti HAV and anti-HBc and in 1 case antibodies anti-HAV and anti-HCV. PMID- 18389723 TI - The establishment of resistance phenotypes for bacteria isolated from outpatients in urine cultures. AB - From 1911 outpatients, who addressed a Timisoara private clinical laboratory, from January to December 2005, we collected 1,889 urine cultures, 431 being positive. Bacteria identification was generally done using morphological, cultural, biochemical characters and pathogenicity tests. Sensitivity testing to antimicrobial medical drugs was done by using the classical diffusion Kirby-Bauer method and the automatic analyzer Osiris, also. The main bacteria involved in the etiology of these infections were represented by Enterobacteriaceae, head of the list being Escherichia coli (81.21%), followed by Klebsiella pneumoniae (8.35%) and Proteus mirabilis (3.02%). We also isolated Gram positive cocci (in a much smaller proportion), mainly represented by Enterococcus faecalis (1.16%), Staphylococcus aureus (0.93%), Streptococcus agalactiae, and also Gram negative non-fermentative bacilli, such as Pseudomonas aeruginosa (0.93%) or Acinetobacter baumanii (0.23%). As soon as we performed the sensitivity tests, we divided them in resistance phenotypes: Most of the Enterobacteriaceae were integrated in the wild phenotype, followed by the penicillinase producing phenotype. An E. coli strain (0.29%) and 3 Klebsiella pneumoniae strains (8.33%) were integrated in the large spectrum, multidrug resistant, beta-lactamase producing phenotype, also associated with resistance to fluoroquinolones and aminoglycosides; Non fermentative bacilli did not present special resistance problems, the four Pseudomonas aeruginosa strains were integrated in the wild phenotype (secreting induced chromosomal cephalosporinase). As for Staphylococcus aureus it was identified a strain having fluoroquinolone resistance, two strains secreting penicillinase and having a K (Nm) phenotype and a strain secreting penicillinase only. Antibiotic resistance represents a major concern for patients, physicians, healthcare managers, and policymakers. The use of antibiotics is closely linked with the development of acquired antibiotic resistance. PMID- 18389724 TI - Virulence genotypes of Escherichia coli strains from bacteremia in relation to phylogeny. AB - Bacteremia is the principal way of dissemination of local infections to distant organs. Escherichia coli bacteremia is almost always clinically significant, suggesting an increased risk of developing sepsis syndrome. Fifty-one E. coli bloodstream human isolates were analyzed using PCR technique for several molecular markers associated with extraintestinal virulence, and their phylogenetic group assignment, taking into account the link between the phylogenetic background and the intrinsic virulence of this species. Sixteen virulence genotypes have been identified, the majority of the blood isolates carrying the association of two genes. The genes encoding type 1 fimbria and aerobactin had the highest prevalence. As a confirmation of other studies, the strains assigned to E. coli phylogenetic group B2 exhibited the highest concentration of virulence genes, and represented almost half of the clinical blood isolates. The multifactorial virulence of E. coli strains isolated from invasive infections reflects a phylogenetic inheritance, and supports the concept of ExPEC pathotype as a subset of E. coli population involved in human infectious diseases. The surveillance of geographical variation of E. coli pathogenic clones is useful for epidemiological analysis. PMID- 18389725 TI - Evaluation of related with age of IFN-gamma levels of treated leprosy patients and their healthy relatives compared with a control group. AB - OBJECTIVE: This study was to search how change with age of IFN-gamma (IFN-gamma) levels of leprosy patients who had received or still undergoing treatment, their healthy close relatives and healthy people. METHOD: From a total of 39 patients, 28 had received leprosy treatment and recovered with some defects [17 Lepromatous Leprosy(LL), 7 Borderline Lepromatous (BL), 3 Borderlien Tuberculoid (BT), 1 Tuberculoid form (TT)] and 11 (9 LL, 2 BL) were undergoing therapy in a hospital at that moment. Close relatives of the patients included 39 people living with treated patients in the same house and the control group consisted of 39 healthy people. Human IFN-gamma kits were used as a solid phase sandwich ELISA method. RESULTS: The IFN-gamma values of treated leprosy patients, therapy receiving patients, relatives group's and control group's were detected as 11-20 pg/mL, 11 18 pg/mL, 12-145 pg/mL and 12-65 pg/mL respectively. CONCLUSION: According to the statistical evaluation of these three groups, IFN-gamma levels of patients were detected lower than other two groups. According to the age, if IFN-gamma levels were compared, it was detected that significant relationship between relatives groups and interferon gamma levels. PMID- 18389726 TI - Immunity status against coxsackie viruses B in 25 patients with acute myocarditis. AB - Viruses are an important cause of myocarditis, particularly the enterovirus group B coxsackievirus. Viral infection may be suspected on the basis of history and presentation and can be proved by direct or serological identification of virus. Twenty-five patients were diagnosed with acute myocarditis and were investigated with a serologic test battery covering Coxsackie viruses group B types 1 to 5 at the National Reference Center for Enteroviruses in Cantacuzino Institute Bucharest, Romania. A possible Coxsakie B virus etiology could be documented in 11 from 25 cases with acute myocarditis and high titers against Coxsackie virus B type 2 (1 patient), type 3 (5 patients) and type 5 (in 4 patients) were detected. In one HIV positive patient (17 years old), a concomitant infection with Coxsackie virus B types 2 and 4 was detected. The earlier detection of enterovirus myocarditis could be followed by antiviral therapies with a potential therapeutic role. PMID- 18389727 TI - Heavy metals resistant plasmid-mediated utilization of solar by Pseudomonas aeruginosa AA301. AB - Solar-degrading bacteria, Pseudomonas aeruginosa strains, were isolated from Egyptian soil by Mineral Salt Medium (MSM) supplemented with Solar (motor fuel) from different oil-contaminated sites in Sohag province. The strain AA301 of Pseudomonas aeruginosa showed appreciable growth in MSM medium containing high concentrations of Solar ranging from 0.5 to 3% (v/v), with optimum concentration at 1.5%. Solar was used as a sole carbon source and a source of energy by the bacterium. The ability to degrade Solar was found to be associated with a single 60-kb plasmid designated pSOL15. The plasmid-cured variant, which was obtained by culturing in LB broth with kanamycin, lost the plasmid indicative the ability to degrade Solar must depend on this plasmid. The wild type isolate, Pseudomonas aeruginosa AA301 and transformant strain, have maximum growth (OD600 = approximately 2) on Solar, however the plasmid-cured variant did not have any significant growth on Solar. Moreover, resistance to a wide range of heavy metals such as Mn2+, Hg2+, Mg2+, Cd2+, Zn2+, and Ni2+ was also 60-kb plasmid-mediated. Therefore, the strain AA301 could be good candidate for remediation of some heavy metals and oil hydrocarbons in heavily polluted sites. PMID- 18389728 TI - The molecular characterization of poliovirus strains by the RT-PCR-RFLP assay and its use in the active surveillance for acute flaccid paralysis cases in Romania between 2001-2006. AB - Poliomyelitis, an acute disease of the the central nervous system can be controlled through the use of inactivated virus vaccine (IPV) or live attenuated vaccine (OPV). The goal of the Global Polio Eradication Initiative is to stop the global transmission of poliovirus. Our study is a retrospective and prospective study that was made because in 2002 was isolated from one acute flaccid paralysis AFP case and eight healthy contacts belonging to the same small socio-cultural group having a low vaccine coverage living in Babadag a town in Romania a serotype 1 vaccine-derived poliovirus (VDPV), recombinant S1/S2/S1. The 67 poliovirus strains isolated in Romania between 2001-2006 from acute flaccid paralysis cases (AFP) (n=20, age = 3 months - 10 years), facial paralysis cases (FP) (n=5, age = 3 months - 3.7 years) and healthy's AFP contacts (n=42, age = 2 months - 5.10 years) were molecular investigated to confirm the vaccine origin of these strains and for the detection of recombinant strains. The identification of these strains was achieved through reverse transcription (RT), polymerase chain reaction (PCR) and restriction fragment length polymorphisme assays (RFLP) applied to two sequences of the viral genome, which are located in VP1-2A (2870 nt - 3648 nt) and 3D (6086nt - 6516 nt) regions. For the strains investigated in VP1-2A region, the RFLP profils after digestion with 3 restriction enzymes (Rsal, Ddel, Hinf l) were Sabin-like only with one exception, VDPV strain. In 3D region after digestion with one, two or three enzymes the genomes of most poliovirus (PV) strains were found to be similar to the original vaccine Sabin strain. In 9 AFP cases the profils detected after RFLP in 3D region were S3/S1 (n=3); S3/52 (n=2); S2/S1 (n=2), S2/S1 + S2 (n=2). In 3 FP cases the profils were S3/S2 (n=2), 53/S1 (n=1). In 11 healthy contacts the RFLP profiles in 3D region were S3/S2 (n=6); S1/S1 +S3 (n=1); S3/S1 + S2 (n=1), S3/S2 + S3 (n=1), S2 + S3/S1 (n=1), S1/S2 (n=1). PMID- 18389729 TI - Diagnosis of toxoplasmosis in pregnancy: importance of immunoglobulin G avidity test. AB - The immunoglobulin G (IgG) avidity test has proved to be a highly useful test in the diagnosis of toxoplasmosis during pregnancy, especially in combination with conventional serological assays. Acute infections at the time of gestation predispose the offspring to the risk of congenital toxoplasmosis. The IgG avidity test was developed to differentiate between recent and more distant infection; this method is valuable in the situation in which a single serum sample is obtained in the first trimester of pregnancy. This paper describes the utility of IgG avidity test during pregnancy, and its role in ruling out, by a high avidity, a recently acquired infection. Testing for specific IgG avidity has been reported to be useful for confirmatory testing in patients who have positive IgG and IgM antibodies. PMID- 18389730 TI - Biochemical changes in rat testis induced in vitro by reactive oxygen species. AB - We report the effects of reactive oxygen species generated by ultraviolet-A radiation on some biochemical parameters specific for oxidative stress, in rat testis homogenates. Results show an increase in lipid peroxidation products under ultraviolet-A exposure, and suggest that the involved mechanism is typical for a radical-mediated chain reaction. The amount of SH groups also increases during irradiation, probably as a consequence of conformational changes in proteins. Electrophoresis results revealed protein pattern changes mainly in the low molecular weight domain. The catalytic activities of alkaline phosphatase and gamma-glutamil transpeptidase are modified under the oxidative conditions generated by reactive oxygen species. The changes of the enzymatic activities are UVA exposure time-dependent, suggesting that conformational modifications are responsible for enzymatic activities enhancement. PMID- 18389731 TI - Critical analysis of the influence of different parameters on the microtensile bond strength of adhesives to dentin. AB - PURPOSE: To evaluate how six different test parameters (TP) of the microtensile bond strength (microTBS) methodology influence the test outcome. MATERIALS AND METHODS: The microtensile bond strength of adhesives bonded to dentin was measured, and the resultant failure patterns and stress-time graphs were analyzed, varying the microTBS specimen-fixation mode (TP1), the microspecimen geometry (TP2), the microspecimen bond surface area (TP3), the remaining dentin thickness of the microspecimens (TP4), the procedure employed to glue the microspecimen to the microTBS jig (TP5), and the microTBS crosshead speed (TP6). RESULTS: The orientation of the microspecimens to the actual interface primarily influenced the resultant microTBS value and the eventual failure pattern. A notched jig consistently yielded significantly higher mciroTBS values than samples fixed onto a flat jig or following a "top-bottom" design (TP1). A higher bond strength, a lower standard deviation/coefficient of variation, a failure closer to the interface, and a more representative number of specimens per tooth were obtained for nontrimmed, square microspecimens (TP2). The most reproducible results were obtained using square specimens of 1 mm2 (TP3), which could also be most easily prepared. The remaining dentin thickness (TP4) was an influential factor as well, whereas the microspecimen gluing protocol (TP5) and the crosshead speed (TP6) appeared less influential. CONCLUSION: As several test parameters significantly influenced the microTBS outcome, sufficient attention should be paid to these methodology variables with regard to the microTBS-test reliability and reproducibility. PMID- 18389732 TI - Dentin bonding on different walls of a class II preparation. AB - PURPOSE: To evaluate the bond strength on different cavity walls of Class II preparations. Different bonding systems and the effect of thermomechanical cycling were investigated. MATERIALS AND METHODS: Human third molars received MOD preparations with dentin margins. Teeth were randomly assigned to 18 groups (n=5) according to the combination of cavity wall (axial, occlusal, and gingival), bonding system (Single Bond Plus, Clearfil SE Bond, and Adper Prompt) and the occurrence of thermomechanical cycling. Restorations were concluded with Filtek Z250 composite. Specimens were sectioned according to the respective cavity wall (4 slabs/restoration), and the adhesive interface was trimmed to an hourglass shape (1 mm2). Slabs were tested under tension, and failure mode was observed. Bond strength data were analyzed with three-way ANOVA/Tukey's test. RESULTS: Single Bond Plus and Clearfil SE Bond performed similarly under most experimental conditions. Single Bond Plus presented similar bond strength on the three cavity walls, regardless of the aging condition. Clearfil SE Bond exhibited significant differences among cavity walls: the occlusal wall showed higher means in both aging conditions. Non-aged gingival walls and aged axial and gingival walls yielded lower means. Non-aged Adper Prompt produced similar bond strengths on the three cavity walls. After thermomechanical cycling, the gingival wall showed lower means. CONCLUSION: The effect of cavity walls was dependent on the bonding system and thermomechanical cycling. Adper Prompt demonstrated bond strengths lower than Single Bond Plus or Clearfil SE Bond under most experimental conditions. PMID- 18389734 TI - Effect of evaporation of solvents from one-step, self-etching adhesives. AB - PURPOSE: To investigate whether and to what extent the bonding capacity of one step, self-etching adhesives is influenced by the degree to which solvent is evaporated. MATERIALS AND METHODS: Seven one-step, self-etching adhesives were tested (Adper Prompt L-Pop, Clearfil S3 Bond, Futurabond NR, G-Bond, Hybrid Bond, iBond, Xeno III). The variation in degree of evaporation was obtained by varying the duration of the air-blowing step. The duration required to immobilize the adhesive layer, as established in a pilot study, was used as control. Two experimental air-blowing durations, shorter (half the control duration) and longer (double the control duration) than the control duration, were chosen. The resin composite Herculite XRV was bonded to flat human dentin surfaces treated with one of the adhesives following manufacturer's instructions, except for the air-blowing duration after application. After being stored in water at 37 degrees C for 1 week, the bonded specimens were broken in shear. Failure modes were evaluated under stereomicroscope. RESULTS: Air-blowing duration and brand of adhesive both had an effect on shear bond strength. An interaction was found between adhesive and air-blowing duration. Some adhesives were insensitive to variations in air-drying duration, but in general, air-blowing durations shorter than the control duration produced lower shear bond strengths. Significant effects of adhesive and air-blowing duration were also detected in relation to failure mode. More adhesive failures were observed with shorter air-blowing durations. A significant negative correlation between number of adhesive failures and bond strength was found. CONCLUSION: On the basis of this in vitro study, it may be concluded that the one-step, self-etching adhesives evaluated were sensitive to degree of evaporation of the solvents. PMID- 18389733 TI - Influence of Er,Cr:YSGG laser treatment on the microtensile bond strength of adhesives to dentin. AB - PURPOSE: In light of the concept of minimally invasive dentistry, erbium lasers have been considered as an alternative technique to the use of diamond burs for cavity preparation. The purpose of this study was to assess the bonding effectiveness of adhesives to Er,Cr:YSGG laser-irradiated dentin using irradiation settings specific for cavity preparation. MATERIALS AND METHODS: Fifty-four midcoronal dentin surfaces, obtained from sound human molars, were irradiated with an Er,Cr:YSGG laser or prepared with a diamond bur using a high speed turbine. One etch-and-rinse (Optibond FL/Kerr) and three self-etching adhesives (Adper Prompt L-Pop/3M ESPE, Clearfil SE Bond/Kuraray, and Clearfil S3 Bond/Kuraray) were used to bond the composite to dentin. The microtensile bond strength (microTBS) was determined after 24 h of storage in water at 37 degrees C. The Kruskal-Wallis test was used to determine pairwise statistical differences (p < 0.05). Prepared dentin surfaces, adhesive interfaces, and failure patterns were analyzed using a stereomicroscope and Field-emission gun Scanning Electron Microscopy (Feg-SEM). RESULTS: Significantly lower microTBS was observed to laser irradiated than to bur-cut dentin (p < 0.05), irrespective of the adhesive employed. Feg-SEM photomicrographs of lased dentin revealed an imbricate patterned substrate and the presence of microcracks at the dentin surface. CONCLUSION: Morphological alterations produced by Er,Cr:YSGG laser-irradiation adversely influence the bonding effectiveness of adhesives to dentin. PMID- 18389735 TI - Masticatory fatigue, fracture resistance, and marginal discrepancy of ceramic partial crowns with and without coverage of compromised cusps. AB - PURPOSE: To evaluate preparation designs of compromised cusps and whether or not they influence masticatory fatigue, fracture resistance, and marginal discrepancy of ceramic partial-coverage restorations (PCRs) luted on mandibular molars. MATERIALS AND METHODS: Sixty-four caries-free molars were equally divided into four groups. Control group NP received no preparation (NP). Group B-IN received a basic inlay (IN) preparation with buccal (B) cusp conservation and occlusal reduction of both lingual cusps. Group B-ON was prepared in the same way, except buccal cusps were prepared with an angle of 45 degrees to the occlusal plane (buccal onlay). Group B-OV preparation was similar to group B-ON, but buccal cusps received a further shoulder preparation on the buccal aspect (buccal overlap). Forty-eight all-ceramic IPS e.max Press PCRs were fabricated and luted adhesively. Specimens underwent mouth-motion fatigue (1.2 million cycles, 1.6 Hz, 49 N) and 5500 thermal cycles (5 degrees C/55 degrees C). Fracture patterns were observed. Surviving specimens were loaded until fracture. Marginal discrepancies were examined. RESULTS: Only one specimen of group B-ON fractured during fatigue. Median fracture loads (N) [IQR = x(.25) - x(.75)]: group NP = 1604 N [1182-1851 N], group B-IN = 1307 N [1262-1587N], group B-ON = 1396 N [817-1750N], group B-OV = 1205 N [1096-1542N]. No significant differences in fracture resistance were found between restored molars and unprepared teeth (p _ 0.18). Different preparation designs showed no significant influence on PCR fracture resistance. Mouth-motion fatigue caused a significanty decrease of marginal accuracy in groups B-IN (p = 0.009) and B-ON (p = 0.008). Marginal discrepancy values of groups B-IN and B-OV were significantly different after fatigue (p = 0.045). CONCLUSION: Ceramic coverage of compromised cusps did not demonstrate an increase of fracture resistance after fatigue when compared to less invasive partial coverage restorations. However, enhanced exposure of restoration margins to occlusal wear could result in more extensive marginal discrepancies. PMID- 18389736 TI - Incisor compliance following operative procedures: a rapid 3-D finite element analysis using micro-CT data. AB - PURPOSE: New methods are available for the rapid generation of 3-D finite element models of dental structures and restorations. Validation of these methods are required. The aim of the present study is to utilize stereolithography and surface-driven automatic meshing to generate models of specific restorative conditions, and to examine these models under loading. The data generated are compared to existing experimental data in an attempt to validate the model. MATERIALS AND METHODS: An intact maxillary central incisor was digitized with a micro-CT scanner. Surface contours of enamel and dentin were fitted following tooth segmentation based on pixel density using an interactive medical image control system. Stereolithography (STL) files of enamel and dentin surfaces were then remeshed to reduce mesh density and imported in a rapid prototyping software, where Boolean operations were used to assure the interfacial mesh congruence (dentinoenamel junction) and simulate different tooth preparations (endodontic access, veneer, proximal, and Class III preparations) and restorations (Class III composites). The different parts were then imported in a finite element software package to create 3D solid models. A 50-N point load perpendicular to the tooth's long axis and centered on the incisal edge was applied either on the buccal or palatal surface. The surface strain was obtained from selected nodes corresponding to the location of the strain gauges in the validation experiments. RESULTS: The increase in crown flexure (compared to the unaltered tooth) ranged from near zero values (conservative endodontic access, removal of proximal enamel) to ca 10% (aggressive endodontic access, conservative Class III preparations), 23% and 34% (moderate and aggressive Class III preparations, respectively), and 91% (veneer preparation). Placement of Class III composite resin restorations resulted in 85% recovery of the original crown stiffness. 3D FEA data correlated well with existing experimental data. In two situations, smaller FEA strains were recorded compared to the experimental strains, perhaps due to enamel cracking under the strain gauges. This artefact was not simulated by the FEA models. CONCLUSION: Experimental data validated the FEA models. The described method can generate detailed three-dimensional finite element models of a maxillary central incisor with different cavities and restorative materials. This method is rapid and can readily be used for other medical (and dental) applications. PMID- 18389737 TI - Coupling of composite resin cements to quartz fiber posts: a comparison of industrial and chairside treatments of the post surface. PMID- 18389738 TI - Rehabilitation of advanced periodontal problems by using a combination of a glass fiber-reinforced composite resin bridge and splint. AB - Rehabilitation of masticatory ability in patients with reduced periodontal tissue support is a multiple challenge in dentistry. It has been shown that tooth mobility is reduced by a splint in place. Splints may be constructed of various materials. Development of fiber-reinforced composite materials offers a new and conservative approach for tooth replacement and stabilization. This case report describes the rehabilitation and 2-year follow-up of a patient with advanced periodontal problems by using a glass fiber-reinforced composite resin material. PMID- 18389739 TI - The $34 trillion problem. PMID- 18389740 TI - Why McCain has the best health-care plan. PMID- 18389741 TI - [Right hemicolectomy for colon cancer: a prospective randomised study comparing laparoscopic vs. open technique]. AB - Since 2001 we have conducted a prospective randomised study of right laparoscopic assisted hemicolectomy vs open right hemicolectomy for right colon cancer in order to assess the differences in intraoperative and postoperative results as well as oncological clearance. Thirty-three patients with right colon cancer received laparoscopic-assisted right hemicolectomy (LRH) and were compared with 33 patients who underwent open right hemicolectomy (ORH). We analysed morbidity and mortality, number of postoperative days of starving, postoperative days tolerating a liquid diet and a soft diet, duration of postoperative ileus, as well as the distance of the resection margin from the tumour (< 5 cm or >5 cm), and the number of lymph nodes found in the resected specimen. We also compared the length of operative time, blood loss, and any associated surgery. Morbidity occurred in 1 patient undergoing LRH (3.0%) as against 4 patients (12.1%) in the ORH group (p < 0.05). Postoperative ileus lasted 3.15 days (range: 3-5 days) in the LRH group vs 3.0 days (range: 1-4 days) in the ORH group. Median operative time was 251 min (range: 130-360 min) in the LRH group vs 222.9 min (range: 135 360 min) in the ORH group, while blood loss amounted to a median of 135 mi (range: 100-300 ml; SD +/- 42.9 mi) in the LRH group vs. 404.1 ml (range: 250 1000 ml; SD +/- 159.3 ml) in the ORH group (p <0.05). The distance of the resection margin from the tumour was more than 5 cm in both groups. In the LRH group a median of 12.7 lymph nodes were removed (range: 9-31; SD +/- 4.5) vs. 18 lymph nodes in the ORH group (range: 8-29; SD +/- 3.9) (p < 0.05). Associated surgery was performed in 15.1% of cases in both groups. In our experience LRH presents a statistically significant advantage in terms of morbidity and blood loss compared to ORH. Equivalent oncological clearance was obtained, fulfilling the stated criteria of 5 cm free resection margins and number of lymph nodes resected, though we removed fewer lymph nodes in LRH compared to ORH (p < 0.05). PMID- 18389742 TI - [Video-laparoscopic excision of pancreatic insulinoma. Experience with 3 cases]. AB - Laparoscopic treatment of lesions of the distal pancreas has gained favour worldwide in the last decade. The objective of this study was to analyze 3 cases of insulinoma successfully treated with the laparoscopic approach. From 2000 to 2007 in our institution 3 patients with insulinoma of the left pancreas were treated with a laparoscopic approach. The insulinoma was diagnosed by helical CT scan, Two cases were treated by left pancreatectomy and one by enucleation. The resections were achieved by laparoscopy with no conversion to laparotomy. There were no intraoperative complications. Average blood loss was 180 mi (range: 150 350). Mean operative time was 232 minutes (range: 225-240). Morbidity consisted in one mild pancreatic fistula after left pancreatectomy that was healed by conservative treatment after 24 days. The mean hospital stay was 13 days (range: 10-20). During the follow-up insulinoma symptoms have disappeared in all patients. This study confirms the feasibility of laparoscopic resection for insulinoma. Operative times were quite acceptable and the conversion rate was nil. Times to oral intake and walking were shorter than after open surgery, as was the mean postoperative hospital stay. PMID- 18389743 TI - [Minimally invasive adrenalectomy: transperitoneal vs. retroperitoneal approach]. AB - Laparoscopic adrenalectomy is now regarded as the procedure of choice for most adrenal glands presenting surgical pathology. The primary adrenal-specific contraindication to laparoscopic adrenalectomy today is the presence of a large adrenal mass with evidence of local infiltration or venous invasion. We used our multicentre experience to compare the transperitoneal (TLA) and retroperitoneal (RLA) minimally invasive approaches. In our study we found statistically significant differences between RLA and TLA in terms of duration of surgery (148 minuti vs. 112; p < 0.005), intra-operative blood loss (439 cc vs 333 p < 0.005; p < 0.005) and time of first oral intake (1.2 +/- 0.5 days vs 1.8 +/- 1.08 days; p < 0.005). The RLA approach is preferable in cases of previous abdominal surgery, but its learning curve is extremely steep. TLA access needs a less demanding learning curve and tends to be faster than RLA, where the working area is penalised by limited manoeuvring space. There is no clear preference between TLA and RLA in the literature. However, the experience of the surgeon still remains the most important variable when choosing between the two approaches. PMID- 18389744 TI - [Endovascular treatment (EVAR) in patients with abdominal aortic aneurysms and synchronous neoplasms]. AB - The association between abdominal aortic aneurysms (AAA) and cancer is becoming more and more frequent, giving rise to several questions regarding the therapeutic and surgical management strategies for both diseases. Endovascular aneurysm repair (EVAR) is the treatment of choice for complex and high-risk patients. In this study we reviewed our experience with patients concomitantly affected by any type of cancer and AAA treated by EVAR at our institution over the last six years. From April 2001 to July 2007, 497 AAA patients underwent open or endografting repair in the 1st Division of General Surgery--Service of Vascular Surgery of the University of Verona. In 53 cases (10.6%) an association with a solid neoplasm was found and 27 of these patients (50.9%) were treated by EVAR. Twenty patients underwent a two-stage approach, with EVAR performed first, while in 5 cases a one-stage approach was preferred on the basis of the general condition of the patients, the site of the tumour to be resected, the logistic possibilities and increased experience of the operators with EVAR. Two patients received chemotherapy after EVAR. There was no in-hospital mortality and four perioperative complications (14.8%) were registered. During a mean follow-up of 25.7 months (range: 2-64 months) 5 deaths occurred, 2 in the short term and 3 in the long term, none of which were related to AAA treatment. Three type-2 endoleaks occurred that sealed spontaneously and 62.9% of the treated aneurysms had a mean 20% decrease in diameter while the others presented no variations. In our experience, EVAR was a safe and effective treatment of AAA patients with concomitant malignancies with a relatively low procedure-related morbidity and no mortality. A simultaneous surgical approach can be achieved safely, performing EVAR as the first step without significant risks. Simultaneous treatment has the advantage of avoiding a second major procedure and eliminates the risk of aortic aneurysm rupture in the postoperative period or during chemotherapy in patients who are usually in poor general condition. Care must be taken with regard to the choice of the device to be used and the possible vascular complications of the visceral circulation. In our opinion, EVAR should be considered the treatment of choice in these patients, taking into account, however, that this treatment is not always feasible in all cases and that in patients with a normal life expectancy (tumour-cured) it may not always be the right choice. Thus, a multidisciplinary approach is necessary in the individual evaluation of these challenging patients in order to make the right decisions. PMID- 18389746 TI - [The thyroid pyramidal lobe: frequency, morphological features and related diseases]. AB - The Authors examine the features of thyroid pyramidal lobe in 1405 operated patients over the period from January 1992 to January 2006. Pyramidal lobe morphology and diseases are reported. The pyramidal lobe was detectable in 25.5% of cases, mostly in women, the primary disease invariably arising from the main gland. The Authors conclude that the pyramidal lobe is an embryological remnant which may be involved in diseases arising from the main gland. PMID- 18389745 TI - [Postoperative metabolic acidosis: use of three different fluid therapy models]. AB - Intraoperative fluid administration is considered an important factor in the management of metabolic acidosis following surgical procedures. The aim of this study was to compare three types of intraoperative infusional models in order to evaluate their effect on acid-base changes in the immediate postoperative period as calculated by both the Henderson-Hasselbach equation and the Stewart approach. Forty-seven patients undergoing left hemicolectomy were enrolled in the study and assigned randomly to receiving 0.9% saline alone (Group A, n=16), lactated Ringer's solution alone (Group B, n=16) or 0.9% saline and Ringer's solution, 1:1 ratio (Group C, n=15). Arterial blood samples were taken before operation (t0) and 30 min after extubation (t1) in order to measure the acid-base balance. The results showed a metabolic acidosis status in Group A patients, whereas Group B exhibited metabolic alkalosis only by means of the Stewart method. No difference was found in Group C between the time points t0 and t1 when using either the Henderson-Hasselbach equation or using the Stewart model. We conclude that saline solution in association with Ringer's solution (1:1 ratio) appears to be the most suitable form of intraoperative fluid management in order to guarantee a stable acid-base balance in selected surgical patients during the immediate postoperative period. PMID- 18389747 TI - [Therapeutic and diagnostic protocol for mild acute biliary pancreatitis: our experience and review of the literature]. AB - Through a critical review of the literature, the authors analyze and re-assess the current diagnostic and therapeutic algorithms used in the treatment of mild acute biliary pancreatitis, reporting their experience with 27 cases observed in the Policlinico Umberto I Emergency Department (Rome) over the period from March 2003 to May 2005. All patients were treated with the same diagnostic and therapeutic protocol: once the diagnosis of acute biliary pancreatitis had been made and the severity evaluated, patients presenting clinical or ultrasonographic signs of main biliary duct stones underwent ERCP within 72 hours of onset of symptoms. All patients then underwent a standard-technique laparoscopic cholecystectomy during the same hospital stay, and whenever ERCP had not been performed preoperatively, an intraoperative cholangiography was performed at the time of surgery. No intra- or postoperative complications were observed, with a mean hospital stay of 10.6 days (range: 5-25 days). PMID- 18389748 TI - [Our experience in the management of patients with Mirizzi syndrome]. AB - The authors report their experience in the management of patients with Mirizzi Syndrome (MS) admitted, over a period of 15 years, at the General Surgery of Emergency Department of Cardarelli Hospital, Naples, Italy. All patients were admitted and surgically treated in emergency save for one. Out of 12 patients, cholecystectomy was performed in 7 cases. In others 5 patients, with cholecystocholedochal fistula, cholecystectomy with positionig of T-Tube was performed in 4 cases (MS-II); finally, 1 patient with MS type III undewrwent choledochojejunostomy. According to literature, the diagnostic protocol included abdominal ultrasonography and CT scan of the abdomen for all patients; in one case, a cholangio-MRI was performed to clarify the diagnosis. The preoperative diagnosis is essential to reduce risk of iatrogenic injuries. The cholangio-MRI, used to this extent, clarifies the site of obstruction, shows the anatomy of the biliary tree and allows to make all the possible differential diagnoses in order to exclude the presence of biliary tumors before surgery. The intraoperative cholangiography remains mandatory to clarify the anatomy of the biliary tree. In the cases we have treated, ERCP was never performed. We believe that ERCP has limited indications and unsatisfactory outcomes for both diagnosis and treatment of MS. Pathological examination of the fresh-frozen surgical specimens was always performed intraoperatively to exclude the presence of concomitant cancer of the gallbladder. The traditional treatment of patients with MS is surgery, as confirmed by our experience. We perform cholecystectomy for MS type I and cholecystectomy with direct repair of the biliary fistula over aT tube for MS type II. Patients with MS type III usually undergo a tailored operation based on the intraoperative findings, while choledochojejunostomy is mandatory for patients with MS type IV. Laparoscopic surgery is indicated only for MS type I and II. It seems to carry a higher risk for the patient and we do not use this approach in the emergency settings. PMID- 18389749 TI - [Indications and limits of laparoscopic treatment for diverticular disease of the colon: personal experience]. AB - The aim of the study was to evaluate the role of laparoscopic surgery in diverticular disease of the colon, in the experience of a specialized centre. Sixty-seven patients were observed from November 2004 to March 2006 at the Robert Koch Krankenhaus of Gehrden (Hannover) with a diagnosis of acute diverticulitis, chronic diverticulitis and/or complications and submitted to elective or emergency surgery. The mean operating time was 171.5 minutes for the laparoscopic approach, and 142.7 minutes for open surgery. Return to normal bowel function occurred after 3.7 days for laparoscopy, as against 4.4 days for open surgery. Mean hospital stay was 9.8 days for the laparoscopic approach and 16.3 days for open surgery. Morbidity was 18.6% (8 cases) in the laparoscopic group and 25% (6 cases) in the open group. Mortality was 0%. Re-operation was necessary in 5 cases in the laparoscopic group (11.6%) and in 4 cases (16.6%) in the open group. Laparoscopy is an important innovation in the surgical treatment of diverticular disease. This approach should be assessed in relation to patient characteristics, medical history and clinical presentation. The advantages of laparoscopy are shorter postoperative hospital stay, less postoperative pain, earlier discharge, better cosmetic result, less blood loss and less peritoneal contamination. In the advanced stages of disease open surgery still remains very important. PMID- 18389750 TI - Effects of high and low ligation on survival in patients operated for colorectal cancer. AB - In spite of numerous studies on the subject, it is still unclear whether or not high ligation of the inferior mesenteric artery (at about 1 cm from its origin) improves the 5-year survival rate in patients operated on for colorectal cancer in comparison to low ligation (ligation below the origin of the left colic artery). From February 2000 to November 2001 40 patients with cancer of the colic segment between the descending sigmoid junction and the low rectum underwent surgical colorectal resection and low ligation of the inferior mesenteric artery. At the end of 5 years of observation we report a survival rate of 70% which is not very far from the value reported in the literature. In our study, the incidence of lymph-node metastases, inexistent in patients with T1 grading increases with the increase in the TNM T grading but does not depend on the location of the cancer. In our patients age below 65 years was a negative prognostic indicator because colorectal tumours in patients of that age are associated with a higher incidence of lymph-node metastases. On the basis of the data we obtained, it is also evident that the 5-year survival rate decreases in proportion to the increase in the distance of the lymph-node metastases from the mesenteric margin of the colon. In conclusion, in the treatment of cancers located between the descending sigmoid junction and the low rectum, we prefer to execute a low ligation of the inferior mesenteric artery because it exposes the patient to a lower risk of intra- and postoperative complications and also because several authors have demonstrated that high ligation with removal of lymph nodes at the origin of the artery for colorectal cancer does not improve the 5-year survival rate. PMID- 18389751 TI - [Recurrent varicose veins due to neovascularisation: can they be prevented?]. AB - The authors analyse the problems of recurrent varicose veins on the basis of their own experience. The 5-year incidence of recurrent varicose veins ranges from 25 to 40% after internal saphenectomy. The recurrences should be classified in relation to the site and pathogenetic mechanism. The authors assess the effectiveness of a prevention method for sapheno-femoral stump recurrences. The study population consisted of 80 patients undergoing internal saphenectomy. In 34 (Group A) a traditional safenectomy was performed with short stripping, while in 46 (Group B) closure of the ostium with a flap of pectineal fascia was performed in order to prevent recurrences caused by neovascularisation. In Group A the recurrence rate was 27.1% as against 14.8% in Group B. Recurrences originating from the sapheno-femoral stump are mainly caused not only by an inadequate echo colordoppler preoperative diagnosis and an inadequate or insufficient crossectomy, but also by neovascularisation. The pectineal flap method in our experience is a very simple, effective and complication-free technique to appreciably reduce recurrent varicose veins originating from the sapheno-femoral stump. PMID- 18389752 TI - [Perineo-ano-rectal injuries: clinical experience]. AB - Traumatic lesions involving the rectum, perineum and anus are infrequent but difficult to treat, requiring experience with trauma and colo-proctological surgery. The aim of the treatment is to repair the lesions and to minimise the early complications which are the main cause of failure and of late complications and disability. The most complicated lesions present problems concerning either the surgical strategy or the surgical timing, both of which are essential for a successful outcome. The Authors analyse their recent clinical experience with 7 patients with complex traumatic lesions involving the rectum, perineum and anus, excluding those of gynaecological/obstetric origin and those not involving the sphincter. They evaluated the clinical history, causes and types of lesions, as well as treatment, complications and outcomes. Five of the lesions were caused by impalement, one by an explosion and one by a motorboat propeller blade. Six of the patients (85.7%) were treated by direct primary repair and one (14.3%) by secondary repair after a previous colostomy. All 7 patients achieved complete recovery of the lesions. Only two cases (28.6%) of early complications and one case (14.3%) of persistent minimal sphincter dysfunction occurred. On the basis of these good results, the clinical experience and the literature, the Authors suggest that these perineo-ano-rectal lesions, though often complex, may often be cured by early surgery, confining colostomy only to particular cases. In addition to experience with trauma and the timing of colo-proctological surgery, a knowledge of all the available surgical options is mandatory to achieve the best results. PMID- 18389753 TI - [Iatrogenic pseudoaneurysms of the peripheral arteries]. AB - The aim of the study was to report our clinical experience with the surgical treatment of iatrogenic pseudoaneurysms of the peripheral arteries. The study is a retrospective review of 101 consecutive patients (52 males, 49 females, mean age 66.2 years, range 33-86), with iatrogenic pseudoaneurysms of the peripheral arteries, surgically treated in a vascular unit from October 1990 to June 2006. Duplex ultrasound scanning was employed to support the clinical findings. The surgical treatment consisted in direct closure with polypropylene sutures and, occasionally, patch angioplasty or bypass. Ultrasound compression was effective in one of 4 small aneurysms (< 2.5). No limb loss occurred. There were 4 wound complications (3.9%), one pulmonary embolism (0.99%), and 3 deaths (2.9%), 2 of which not related to vascular repair and one secondary to femoral endoarteritis and septic shock, unrelated to previous implantation of a percutaneous femoral closure device. Although iatrogenic pseudoaneurysms of the peripheral arteries are rarely observed in clinical practice, a significant number of peripheral artery complications may occur after cardiac catheterisation and coronary angioplasty. Failure of conservative treatment requires a traditional surgical repair. The results of our series included a significant mortality rate (2.9%), resulting from the severity of cardiac disease in 2 cases and from the vascular repair itself in one case (femoral endoarteritis). These results substantiate the common observation that patients who require surgery for an iatrogenic pseudoaneurysm are often affected by advanced cardiovascular disease and are liable to suffer the occurrence of complications, with a high risk of death. Therefore, any surgical treatment should be performed with strict adherence to sound vascular surgical principles. PMID- 18389754 TI - [A new accessory for the isostatic anal retractor]. AB - The authors present the characteristics and the advantages deriving from the use of a new accessory to be fitted on the isostatic anal retractor in order to further improve visualisation of the operative field during the treatment of pelvic and anal pathologies. The isostatic anal retractor consists of a system of rotating rings and valves which are positioned with surgical stitches on the patient's buttocks. The new accessory is characterised by one or more elastic systems that can be fixed to the retractor metal ring and attached by means of curved needles to flaps of anal mucosa, affording enhanced visibility of the operative field. A total of 25 patients--15 with mucosal prolapse, 5 with haemorrhoids and 5 with anal fistulas--were treated surgically for their respective pathologies using the isostatic anal retractor and the new accessory. None of the patients observed complained of complications related to the use of the anal retractor or the accessory. In 95% of patients submitted to the procedures using the new accessory, visualisation of the operative field was improved as compared to the use of the anal retractor alone. In 5 patients it was possible to use the new accessory advantageously applied only to the external ring of the retractor. Our operative team judged the use of new accessory to be satisfactory in all cases in which greater visibility of the operative field is required. Use of the new accessory use in association with the isostatic anal retractor proved efficacious for achieving better therapeutic results with the treatments performed, permitting greater operator autonomy and enhancing visualisation of the operative field with respect for sphincter function. PMID- 18389755 TI - Transcystic videolaparoscopy for choledocholithiasis with holmium: YAG laser lithotripsy. A case report. AB - Randomised prospective studies have shown that single-stage management of cholecysto-choledocholithiasis yields results equal or superior to sequential treatment, with a lower incidence of complications primarily associated with choledochotomy. We report the first case of transcystic Holmium:YAG laser lithotripsy using a flexible ureteroscope. The method was found to be a valuable aid in reducing the percentage of choledochotomies when calculi are too large to be retrieved from the common bile duct with normal graspers. PMID- 18389756 TI - [Proctological surgery: use of an absorbable haemostatic sponge]. AB - The aim of this study was to demonstrate the advantages and efficacy of an absorbable haemostatic sponge for use in coloproctology during surgery for the most frequent pathologies of the anal canal and perineal region. The haemostatic sponge is made of absorbent gelatin that is insoluble in water and is endowed with haemostatic power. It can be produced in various sizes and forms adaptable to different uses and presents no toxic properties. The authors tested the haemostatic sponge in a series of consecutive patients referred to 5 coloproctological centres where they were submitted to surgery for pathologies of the anal canal and perineal region. A total of 100 consecutive patients, 50 with a clinico-instrumental diagnosis of grade III-IV haemorrhoids, 20 with a diagnosis of chronic anal fissure, 20 with symptomatic rectocele and 10 with simple fistulas, underwent surgery for their respective pathologies. In all patients treated, at the end of the procedures, the haemostatic sponge was used, achieving optimal haemostatic control of surgical wounds with no infectious complications. Use of the haemostatic sponge proved easy and satisfactory as regards stability in the anal canal and control of haemostasis after surgery for the most frequent proctological pathologies. PMID- 18389757 TI - [Traumatic hernia of the abdominal wall: a case report]. AB - Traumatic hernia of the abdominal wall is a rare type of hernia. A high-energy, localised impact causes the disruption of the abdominal wall musculature without penetrating the skin. The most frequent cause of the trauma is the lower abdomen striking the end of bicycle handlebars in young adults and children. A high level of clinical suspicion is required for diagnosis because of the high rate of associated injuries. The authors report a case of handlebar hernia managed successfully by surgical repair. PMID- 18389758 TI - Laparoscopic resection of a large GIST of the stomach: is it preferable in elderly patients? A case report. AB - Gastrointestinal stromal tumour (GIST) of the stomach is extremely rare in the elderly. Surgical resection of the stomach by partial gastrectomy or wedge resection is the standard treatment. Today the resection can also be performed laparoscopically, especially in the case of small tumours as well as for larger GIST though there are unclearly defined oncological limits. The authors report the successful treatment of a large 7.5 cm GIST of the stomach by laparoscopic wedge resection in a 78-year-old patient. The GIST was almost entirely located intraperitoneally between the stomach and the spleen and could be radically resected with a minimal touch technique. The patient recovered promptly and manifested no recurrence at a 2-year follow-up. The authors focus on the main factors supporting the indication for laparoscopic resection of large gastric GIST, especially in the elderly. The surgical risk/benefit ratios of the different approaches, the surgeon's skills in laparoscopically respecting the rules of oncological surgery, and informed consent of the patient in relation to the limited scientific evidence concerning the main risk factors of recurrence are all important considerations. PMID- 18389759 TI - [Retroperitoneal giant schwannoma: a case report and review of the literature]. AB - Schwannomas are rare tumours that originate in the neural sheath and account for only a small percentage of all retroperitoneal tumours. They are usually solitary, circumscribed and encapsulated lesions eccentrically located on proximal nerves or spinal nerve roots. Presentation is typically varied and non specific, ranging from abdominal pain, an abdominal mass or an incidental finding. The preoperative diagnosis is difficult and laboratory tests are usually unremarkable. We report the case of a 66-year-old female presenting with abdominal pain in her left flank and with an ultrasonographic diagnosis of a left kidney mass. She was diagnosed as suffering from a giant retroperitoneal schwannoma after surgical exploration and complete excision. The role of CT scan and CT-guided needle biopsy is emphasised, in that ultrasonography and fine needle aspiration alone do not provide sufficient information regarding aetiology and malignancy. Radical surgical excision is curative but recurrences may occur. Careful follow-up is needed. PMID- 18389760 TI - [Aorto-oesophageal fistula due to an aortic endovascular prosthesis: case report and state of the art]. AB - Aorto-oesophageal fistula is a rare and often fatal condition due to pathologies of the aorta and oesophagus. Recently, a new aetiology for aorto-oesophageal fistulas has been detected, namely, decubitus of an aortic endovascular prosthesis positioned in the presence of aneurysms. The symptoms are those of Chiari's triad: (1) chest pain and/or dysphagia (2) haematemesis (3) massive haematemesis. If the patient is haemodynamically stable the gold standard diagnostic examination is a CT scan with contrast medium. Aorto-oesophageal fistulas are characterised by a rapidly worsening acute clinical presentation and high postoperative morbidity and mortality. The treatment of aorto-oesophageal fistula is divided into three progressive steps: (1) control of bleeding; (2) prevention of mediastinitis; (3) oesophageal repair. We report a case of a 59 year-old male patient with an aorto-oesophageal fistula due to the decubitus of an endovascular aortic prosthesis previously positioned for a traumatic aneurysm of the descending aorta. We controlled the bleeding in emergency with a Sengstaken-Blakemore tube. Since the cardiovascular surgeons excluded any intervention, we executed a bipolar oesophageal exclusion in our department of general surgery and subsequently positioned a self-expanding oesophageal prosthesis by a retrograde route. The patient survived for 7 months, the cause of death being septic shock. PMID- 18389761 TI - Cystic adventitial disease of the popliteal artery. A case report and review of the literature. AB - Cystic adventitial disease is a rare form of non-atherosclerotic stenosis and one of the main causes of claudication in young and middle-aged men. Approximately 200 case reports are available in the literature to date. It is generally located in the popliteal artery, although it may be found in other arteries and even in veins. The aetiology is still unclear: most authors believe that the cyst may originate from a synovial ganglion close to the adjacent hip joint capsule. Patients affected by adventitial cystic degeneration are often young male non smokers with intermittent calf claudication. The treatment of choice is surgical removal of the cyst or reconstruction with autologous vein or synthetic graft interposition. Percutaneous US-guided cystic aspiration is a recent easy and safe alternative method for treating the disease but may result in local recurrence. We report the case of a 51-year-old male patient with clinically intermittent claudication of the right leg. The arteriogram showed complete occlusion of the right proximal popliteal artery and no evidence of atherosclerotic disease in other vessels. The diagnosis was made at the time of surgery. Surgical exploration revealed a gelatinous material involving the popliteal artery. It was excised and evacuated and a segment of greater saphenous vein interposed. Ultrasound examination 12 months later showed graft patency and absence of local recurrence. PMID- 18389762 TI - Pleomorphic hyalinizing angiectatic tumour of the mesorectal soft tissue. A case report and review of the literatre. AB - Pleomorphic hyalinizing angiectatic tumours are rare stromal lesions histologically resembling both neurilemoma and malignant fibrous histiocytoma and occurring in the subcutaneous soft tissue of the lower and upper limbs and, less frequently, in the chest wall. The case reported here is one of 22 cases published in the medical literature and describes a pleomorphic hyalinizing angiectatic tumour which was localized in a body cavity and developed in the pelvis. The present report is the first of its kind to date. A 53-year-old asymptomatic woman was treated via an open laparotomy. The lesion arising from the left mesorectal tissue was entirely resected. The postoperative course was uneventful and the patient was discharged on postoperative day 3. The patient is still disease-free 58 months after the operation. A review of the literature shows that pleomorphic hyalinizing angiectatic tumours are locally aggressive. A 20% recurrence rate has also been observed in long-term follow-up. These patients should therefore be treated by wide local excision and require long-term surveillance. PMID- 18389763 TI - [Iatrogenic pathology--obscure issue in medical care and neglected problem in medical education]. PMID- 18389764 TI - [Iatrogeny and the elderly]. AB - Discussions about the impact of iatrogeny over general health were avoided and neglected for a very long time. It was brought to medical interest due to recent studies concerning therapeutic compliance, patient's medical education and acute awareness of ethical implications of medical activities. Even if iatrogeny was always present in medicine, we realized but recently the dimensions and the necessity of a coherent and systematic approach of this problem. Iatrogenic pathology in elderly population has even a bigger impact due to conjugation of two major demographic phenomena: an absolute and percentage increase of elderly population parallel with an increase of prevalence of iatrogenic pathology with age. That's why iatrogeny in elderly becomes a major health problem, unfortunately often ignored, neglected and even obscured inspite of well recognized impact over duration and quality of patient's life as well as over the cost of medical assistance. PMID- 18389765 TI - [Eosinophilic esophagitis--from a rare pediatric disease to the forefront of adult gastroenterology]. AB - Eosinophilic esophagitis (EE) is a disease whose presence has exploded in clinical practice. Because of its relative novelty, the epidemiology, pathogenesis, clinical manifestations, diagnosis and treatment of EE are constantly evolving. EE was first recognized as a disease predominantly of children, but the publications in the last few years describe it like an adult disease, too. It is not known if its frequency is truly increasing in an antigen underexposed society or the disease was previously not recognized. The diagnosis requires the histologic finding of more than 20 eosinophils per high powered field in esophageal squamous mucosa. The specific antigens that initiate EE are unknown, but both food and aeroallergens are involved. The mechanisms by which EE induces esophageal dis-motility, chronic inflammation with fibrosis, and stricture formation are still unclear. The mucosa of the patients affected by EE contains increased numbers of CD8 lymphocytes, tumor necrosis factor alpha, I1-5 and eotaxin. The main symptoms in adult are: dysphagia and food impaction. Endoscopy commonly reveals strictures, mucosal rings, linear furrowing, corrugation, and white plaques. The common treatment regimens in children and adults involve the ingestion of topical corticosteroids. PMID- 18389766 TI - [Differential diagnosis of febrile exanthema]. AB - Febrile exanthema (FE) is an extremely polymorphous clinical entity, frequently seen in daily clinical practice. FE is characterized by diffuse rash and fever. FE is classified in 4 types, depending on the primary skin lesion: macular, maculopapular, vesicular, and bullous exanthema. It is of infectious and non infectious cause. Among the infectious causes the most frequent is the viral one. FE may affect all ages, but especially children and young people. Usually, FE raises important issues of differential diagnosis, because its clinical and etiological complexity. To decide the most appropriate therapeutic and prophylactic measures for FE, it is important to know the clinical criteria and the specific diagnostic methods. PMID- 18389767 TI - [Image-guided diagnostic percutaneous puncture biopsy]. AB - Percutaneous aspiration and biopsy have an increasing diagnostic role. Ultrasound and CT are the imaging methods most frequently used for guiding, but fluoroscopy and MRI are also considered. The paper reviews the indications, contraindications, method of ultrasound-guided and CT-guided punctures, and the results of needle aspiration and biopsy for cerebral, head and neck, thoracic, abdominal and musculoskeletal lesions. PMID- 18389768 TI - [Clinical extraintestinal manifestations in patients with ulcerative colitis]. AB - Ulcerative colitis (UC) is a chronic disease clinically manifest either by bowel symptoms alone or extraintestinal symptoms. MATERIAL AND METHOD: Our prospective study included 635 patients with ulcerative colitis (334 males and 301 females, mean age 37.54 +/- 13.84, range 20-70 years). The presence of the common extraintestinal symptoms (ES) was analyzed. RESULTS: Of the 635 investigated patients, these symptoms were found in 83 (13%, 49 males and 34 females, mean age 41.6 +/- 13.95 range 21-70). Patients with ES suffered longer from UC on the average, that is 60.6 years. Most commonly ES involved the joints, 38 (45.8%) patients, hepatobiliary, 28 patients (33.7%), skin, 10 patients (12%) and eyes, 7 patients (8.4%). In 18% of the patients two or more ES were present. ES were clinically detectable after the intestinal symptoms in 81% patients. An increased tendency of ES to occur in patients with a more extensive disease was noticed. CONCLUSION: The prevalence of ES in the UC patients from NE Romania is in agreement with data from other countries. The number of ES supports the need for complex follow-up in these patients. PMID- 18389769 TI - [Difficulties in the treatment of pulmonary tuberculosis in children]. AB - The management of pulmonary tuberculosis in children in Romania is done according to ERS, WHO and IUATLD recommendations, Iasi area being included since 1998 in the pilot study. MATERIAL AND METHOD: We analyzed in a descriptive and retrospective study 254 children with pulmonary tuberculosis admitted to the Iasi Department of Pediatric Lung Diseases in the interval January 2000 - December 2005. The surveyed parameters included: treatment course, side effects of antituberculous drugs, and an assessment of the cases at the end of treatment. RESULTS: This analysis revealed: 98.8% were new cases, 47.2% had received four drug regimen for the first 2 months, 46.9% 3-drug regimen and 5.9% individualized treatment. Of the four-drug regimen 92.5% cases were treated with ethambutol. Of the 5.9% with individualized treatment 12 cases had tuberculosis and HIV/AIDS, out of which 2 with MDR tuberculosis, and 1 case with virus B hepatitis; all of them had fluoroquinolones added as well as a longer period of treatment. Side effects of antituberculous medication were encountered in 4.1%, most frequent being toxic hepatitis (2.5%). Patient evaluation at the end of treatment was done according to WHO recommendations: treatment was completed in 70.5% of cases, cured: 23.6% of the cases; relapses in 2.8%, deceased in 2.8% cases. CONCLUSIONS: Pulmonary tuberculosis treatment in children is difficult and must take into consideration their age and nutrition status as well as the side effects and drug combinations. PMID- 18389770 TI - [Age-related prognostic factors and treatment results for advanced non-small cell lung cancer (NSCLC)]. AB - There is much debate whether the choice of management for elderly patients with locally advanced, inoperable NSCLC with good performance status (PS) should be made entirely on terms of age. MATERIAL AND METHOD: We compared the results of chemotherapy (CT), radiotherapy (RT) and chemoradiotherapy (CT-RT) of the elderly with those of younger patients, and studied predictive factors for survival in these age groups. 231 patients with advanced NSCLC were included in this study, being grouped by age: > 65 years (148p, 64.1%) versus < 65 years old (83p, 35.9%). Univariate analysis and Cox regression models were used to assess significance of variables for prediction of survival. RESULTS: Mean overall survival (OS) for the whole group was 11.48 months (median 9.0 months); no significant difference in median OS was found between elderly and younger patients (11.55 vs. 11.65 months, p = 0.537). Univariate analysis revealed significant survival benefits in both age groups as per weight loss < 5%, absence of comorbidities, earlier clinical stage, high initial Hb levels, longer time-to progression (TTP) and good PS. Treatment type had different survival impact on the age groups; best median survival in patients > 65 years (14.0 months) was obtained by CT alone, while patients < 65 years benefited more (13.35 months) from sequential CT-RT. Logistic regression model identified 5 variables to be significant for survival in all patients: PS, extent of disease, hemoglobin (Hb), TTP and age. When applied to the elderly group, only 4 variables had predictive value: extent of disease, Hb, TTP and presence of comorbidities. Treatment toxicity did not differ significantly between age subsets, except for renal toxicity, which was greater in elderly patients. Age should not be a choice limiting item for the treatment of advanced NSCLC. CONCLUSION: An active therapeutic approach, such as chemotherapy, can be feasible, effective and well tolerated in selected elderly NSCLC patients with a good PS and no associated comorbidities. PMID- 18389771 TI - [Thyroid ultrasonography. Interobserver and intraobserver variations]. AB - Thyroid ultrasonography is a simple diagnostic method for assessing the prevalence of goitre in children, parameter useful in the estimation of iodine supply. The normal limits recommended by WHO and ICCIDD (following the ThyroMobil study) have been debated by studies which found smaller values, even in regions included in the ThyroMobil study. AIM: To assess the importance of the inter- and intra-observer differences in the determination of thyroid volume in children. MATERIAL AND METHODS: Thyroid volume in 30 children was estimated using the rotating ellipsoid formula by 3 different observers. One of them determined twice the volume in 25 children. RESULTS: Mean interobserver variations were 9.51 +/- 8.8%. More important differences were between observer 1 and 3. The differences between every diameter were between 12 and 16%, more important for the longitudinal one. The intraobserver differences were 6.29 +/- 6.12%. We did not find a relation between the value of the thyroid volume and the interobserver (r2 = 0.13, p > 0.05) or intraobserver (r2 = 0.15, p > 0.05) differences. CONCLUSIONS: Interobserver variation may be important, and can partially explain the disagreements on thyroid volume in children. Intraobserver differences, although present, are less calculated. It is important to establish standards for each geographical area and, as much as possible, by the same person. In order to obtain comparable data the techniques of the ultrasound volume evaluation should be harmonized. PMID- 18389772 TI - [Android obesity and breast cancer risk]. AB - The 1998 report on the occasion of the 4th anniversary of International obesity Task Force emphasized that overweight (BMI > 25) is also linked to the increasing incidence of some neoplasias--breast, colon, endometer, prostate or kidney. Carmichael and Bates showed an inverse relationship between BMI and perimenopausal breast cancer. MATERIAL AND METHOD: We studied a group of 110 patients divided in three subgroups--30 patients who had just been diagnosed with breast cancer; 50 patients with breast dysplasia diagnosed by mammography and mammary echography; 30 patients with no modifications on mammography or breast echography. Height, weight, abdominal and hip perimeter have been measured. All have performed complete lipidogramme as well as serum estradiol. RESULTS: 15 of the 30 breast cancer patients (50%) were obese with a WHR > 0.8 compared to 23 (46%) in the mastosis group and 20 (66%) in the reference group. The prevalence differences were not significant, but the association between abdominal obesity and increased serum estradiol is highly significant for group I compared to the other groups (p < 0.01). CONCLUSION: Our study confirms that android obesity and estrogens are linked to breast carcinogenesis promotion by synergic or common pathways. PMID- 18389773 TI - Role of multi-component functional foods in the complex treatment of patients with advanced breast cancer. AB - Cancer increases free radicals and that certain diets can influence oxidative stress. There is little evidence about the functional effect of multi-component functional foods on oxidative stress in breast cancer. MATERIAL AND METHODS: 34 female patients with histologically confirmed breast cancer, stages IIIB and IV were enrolled in a randomised controlled trial. All patients filled out the Quality of Life questionnaire FACT-B version 4, translated in Romanian and validated, obtained from Evanston North-Western Health Care, Illinois, USA. Seventeen patients, mean age 57.84 +/- 11.2, were put on a special diet consisting of 15 ml/day balsamic vinegar from apples and honey, with seabuckthorn berry rich in carotenoids, rosemary, sage and basil extracts, to be used in salads and vegetable soups, 150 g/day whole wheat bread with 2.5% of the nutraceutic mixture VITAPAN, 15 ml/day grape seed extract rich in polyphenols (gr. 1). Seventeen patients served as control (gr. 2). Parameters of the metabolic syndrome were measured: serum lipids, IR-HOMA insulin resistance index. Oxidative stress was measured by the FORT (free oxygen radical) test. Total hydro and liposoluble antioxidants (ACW, ACL) in serum were measured by chemo luminometry. All investigations were done at entry and after 3 months. RESULTS: Student's t test was applied to compare the means within group and between the 2 groups. The Physical well being subscale score of the QOL FACT-B version 4 questionnaire showed a significant difference between the 2 groups (p = 0.001). Also, functional well being subscale (p = 0.004), FACT-G (p = 0.003) and FACT-B (p = 0.002) showed significant differences on comparison. Breast Cancer Score (Additional Concerns) had a borderline significant difference (p = 0.057). Social/Family Well being subscale and Emotional well being subscale scores showed no significant difference. At inclusion, radical activity > 310 FORT units, relevant for increased oxidative stress were present in 95.1% cases. After 3 months radical activity > 310 FORT units were present in 52.8% cases in gr. 1. CONCLUSION: A proper design of foods enriched with bioactive compounds that can lead to significant effects on health represents a promising adjuvant treatment in patients with advanced breast cancer, due to its contribution in lowering the high oxidative stress present in these patients. PMID- 18389774 TI - [Utility of investigations by family physician of some biological parameters in chronic alcoholism]. AB - AIM: To assess some biological alterations induced by chronic alcohol consumption and its correlation with the ingestion period and severity of consumption. MATERIAL AND METHOD: The study was carried out on a series of 98 individuals (age range 20 to 80 years), followed up for alcohol consumption; 45 age-matched persons which did not consume alcohol served as controls. In both series the following parameters were assessed: blood count, mean erythrocyte volume (MEV), platelet count (PC), mean platelet volume (MPV), gamma-glutamyl-transferase (GGT), alanine and aspartate transaminases (ALT, AST). RESULTS: The male/female distribution was 0.98; the average amount of alcohol consumed the last five years: within accepted limits (66.32%), moderate (16.32%), increased (17.36%). The results (average values) revealed, compared to the controls, an increase in MEV from 90.312 to 97.125 I3, normal values for PC and MPV, higher levels of AST and ALT, especially in the heavy drinkers, and a significant increase of GGT level. CONCLUSIONS: Chronic alcohol consumption induces the appearance of important alterations of some biological parameters, in correlation with the consumption period and alcohol amount. A systematic assessment of all of these biological parameters, carried out by the family physician, may be a useful tool in the early detection of severe alterations caused by excessive alcohol consumption. PMID- 18389775 TI - [The significance of neuroimaging in the evaluation of the newborn with neonatal asphyxia]. AB - This is a retrospective study of 3069 newborns admitted to the Neonatology Department of the 1st Obstetrics and Gynaecology Clinic of Cluj-Napoca, Romania. MATERIAL AND METHOD: We considered 38 newborns with the diagnosis of neonatal asphyxia. The neuroimaging performed was transphontanellar ultrasonography, CT scan, MRI, depending on case. Post asphyxic syndrome evaluation, complete neurological examination, EEG and neuroimaging were performed. RESULTS: The incidence of neonatal asphyxia was 1.15%. Neurological signs were present in 63.15% of the cases. Neurological aspects of newborns were present in different grades of hypoxic-ischaemic encephalopathy. EEG signs were present in 33.3% preterm newborns and 50% newborns at term. Transphontanellar ultrasonography showed different grades of intraventricular hemorrhage, periventricular leukomalacia in 35%. CT-scan was performed in 3 cases. MRI evaluation was performed in 8 cases and showed different types of cerebral ischaemia and cerebral atrophy. Neuroimaging is a marker for the evolution of cerebral injury. CONCLUSION: The evaluation of the newborn with neonatal asphyxia must be complex, and include clinical data, EEG and neuroimaging. PMID- 18389776 TI - Multicystic mesothelioma--a rare case of ascites: case report. AB - We present the case of a 37-year-old male, admitted to our clinic with abdominal tenderness, right supraclavicular tumour, and ascites. The presence of ascites was incidentally reported 6 years before, but no other evaluation was done at that moment or during this period. Abdominal ultrasound and CT scan revealed moderate ascites, perivascular adenopathies, and multiple abdominal cystic lesions, while thoracic CT scan revealed the same lesions in mediastinum. Laboratory data were within normal limits, including the tumoral markers, and the tests for hydatid cysts. A biopsy from the right supraclavicular nodule was performed, and based on usual and immunohistochemical stains (calretinin, mesotheline, CK 5/6, CK 7, CK18 diffusely positive in mesothelial cells, and CEA M, bcl-2 and vimentin negative), suggested the diagnosis of mesothelioma. Based on these results, the diagnosis of "multicystic mesothelioma" was made. The patient was referred for surgery. PMID- 18389777 TI - [Adult onset Still's disease with normal level of serum ferritin]. AB - We present a case of an adult onset Still's disease: a 51 year old men presented with one month history of high spiking fever, asymmetric migratory polyarthritis and a previous history of pharyngitis. The diagnostic was based upon clinical criteria and laboratory findings, and necessitated the exclusion of infectious, neoplastic, and other "autoimmune" disease. The systemic involvement in our case induced us to comment therapy with corticosteroid. Patients with systemic disease have a favorable prognosis, with only rare serious complications from the disease (pericarditis, tamponade, diffuse intravascular coagulation, amyloidosis, hepatic disease, and respiratory failure) or the treatment (infections, gastrointestinal bleeding etc.). PMID- 18389778 TI - [Electrocardiographic changes in acute organophosphate poisoning]. AB - Cardiac complications often accompany poisoning with organophosphates. These may be serious and often fatal, being represented by cardiac arrhythmias, electrocardiographic abnormalities and conduction defects, as well as myocardial infarction, a rarely reported complication of acute pesticide poisoning. The extent and pathogenesis of cardiac toxicity from these compounds is not yet clearly defined. We report the case of a 57-year-old woman who presented to our emergency department with coma and acute non-cardiogenic pulmonary edema, as a result of organophosphates ingestion. She was resuscitated for asystole presented shortly after admission; prolonged QTc interval, ST-T changes, right bundle branch block, ventricular tachycardia were recorded. Finally she developed acute anteroseptal myocardial infarction and died despite serum cholinesterase normalization. We believe that admission in an intensive care unit, careful electrocardiographic and enzymatic monitoring of all patients is important for the diagnosis and treatment of cardiac complications of organophosphates poisoning. PMID- 18389779 TI - [Cardiac failure in the newborn. Diagnostic pitfalls]. AB - Echocardiography commonly represents the diagnostic clue in neonatal heart failure (HF). Congenital heart diseases are the most frequent causes of HF in this age group. Arterio-venous malformations are the most common noncardiac causes of HF. Normal cardiac structural findings on echocardiography require further investigations in order to exclude other causes of HF. We present three male patients admitted in the interval 2003-2007 with neonatal HF, systolic murmur, cardiomegaly, normal cardiac structure on echocardiography and intracranial bruit. All three cases were diagnosed with vein of Galen aneurysmal malformation (VGAM) by head ultrasound. According to age and malformation type, different presentation patterns were noticed: early neonatal intractable HF mimicking aortic coarctation, postnatal HF stabilized by drug treatment, and chronic HF in a VGAM with tendency to spontaneous regression. Both head ultrasound and cranial auscultation are mandatory in newborns or infants with no cardiac primary cause of HF. PMID- 18389780 TI - [Dermabrasion in two cases of tuberous sclerosis]. AB - The authors made an attempt at treating tuberous sclerosis (Bourneville's disease) by means of dermabrasion. The technique and postoperative course are described. The immediate results were very encouraging, but in time returned to an appearance close to the initial one. In conclusion, dermabrasion is not an appropriate treatment for tuberous sclerosis. PMID- 18389781 TI - [Echocardiographic evaluation of left ventricular diastolic dysfunction in patients with diabetes mellitus]. AB - The presence of diabetic cardiomyopathy, without any sign of hypertension, valvular or coronary artery disease, is related to diastolic dysfunction, an early sign of diabetic heart muscle disease. Doppler echocardiography evaluation should be performed for all patients with type 2 diabetes mellitus and microalbuminuria, in order to obtain a better management of this metabolic disease. PMID- 18389782 TI - [The ostomy support team. A reality for ostomates]. AB - Stoma is a Greek word meaning mouth or opening. There are many types of surgical stomas and they may be raised on many areas of the abdominal wall. A stoma may be temporary or permanent, may be needed in any age group and may be sited on any part of the abdomen. The specific digestive pathology that could have as result of the surgical management a stoma is represented by colon, rectal and anal cancer, diverticular disease of the colon and rectum, Crohn's disease, ischaemic bowel, volvulus, trauma, Hirschprung disease, imperforate anus, fecal incontinence. This paper aim is to asses the management of fecal stomas and the necessity of a trained ostomy support team. PMID- 18389783 TI - [Long-term results after surgery for colorectal adenocarcinoma, stage I-III. Problems of prognosis]. AB - Colorectal cancer is one of the leading causes of cancer-related death worldwide. STUDY DESIGN: Prospective study on 142 consecutively cases with stage I to III colorectal adenocarcinomas (TNM AJCC/UICC) in which patients underwent potentially curative surgery in one single public health service (1st Surgical Clinic Iasi, Romania) between 2004 and 2005. MATERIAL AND METHOD: The mean follow up was 23.26 +/- 9.78 months (range 2 to 42 months). There were 85 men (59.9%) and 57 women (40.1%) with mean age 63.38 +/- 11.84 years (range 28 to 88 years). The surgical procedures performed were the following: right colectomy (n = 54; 30%); transverse colectomy (n = 2; 1.4%); left colectomy (n = 19; 13.4%); segmental colon resection with anastomosis (n = 5 ; 3.5%); Hartmann procedure (n = 18; 12.7%); anterior rectal resection (n = 11; 7.7%) and abdominoperineal resection (n = 33; 23.2%). With regard to postoperative adjuvant therapy most patients were given chemotherapeutic agents such as 5-fluorouracil and folinic acid. The mean overall survival (months) and 42-months survival rates were calculated. The patients were censored in the survival calculation (Kaplan-Meier method) and Cox regression if they were alive at the endpoint of the follow-up. Some patients were censored because they were "lost to follow-up". Statistical significance is p < 0.05. RESULTS: The factors with a significant negative influence in overall survival and 42-months survival rates were: the age over 70 years, the emergency surgery related to cancer's complications, the advanced AJCC/ UICC stage, vascular invasion, perineural invasion, the recurrence of disease, the moderate and lower differentiated adenocarcinoma and incomplete or not performed chemotherapy. CONCLUSION: Even with a radical surgical approach the advanced stage of colorectal adenocarcinoma has a low prognostic, but some other factors have also a high significance in postoperative outcome. Related to other prognostic factors we performed a review of literature. PMID- 18389784 TI - [The impact of patient and tumor related prognostic factors on survival in non medullary differentiated thyroid cancer. A study of 125 cases]. AB - The aim of the study was to assess the impact of patient--(age) and tumor- related factors (size, extrathyroidal invasion, distant metastasis, multicentricity and lymphatic metastasis) on survival of patients with differentiated thyroid cancer (DTC). MATERIAL AND METHOD: A clinical retrospective study was carried out on a series of 125 patients operated for non medullary DTC in the IIIrd Surgical Unit, in the period 1990-2005. The disease specific survival (DS) was calculated using the Kaplan Meyer method and Cox regression univariate and multivariate analysis was used to assess the impact of prognostic factors on DS. RESULTS: The actual DS at 5, 10, and 15 yrs was 81.3%. Clinicopathological factors significant on univariate and multivariate regression were age over 45 yrs (p = .01), tumor size > 4 cm (p = .00), macroscopical extrathyroidal invasion (p = .000) and distant metastasis (p = .000). PMID- 18389785 TI - [The importance of diurnal pressure curve in evaluation of glaucoma patients]. AB - The assessment of intraocular pressure diurnal curve is not a routine examination. In some situations it might be useful. Determining real intraocular pressure peaks during the nictemeral rhythm can orient the diagnosis to better estimate medication efficiency. MATERIAL AND METHOD: We selected for this study 27 normal patients (52 eyes) and 29 glaucomatous patients (56 eyes). We chose to determine intraocular pressure every three hours, excluding the 3 a.m. in the night determination, thus obtaining a 7 values curve. RESULTS: We compared the data obtained with 5 values curve, excluding the 6 a.m. and 12 p.m. determinations. Analyzing data we observed that there was no statistical difference between pressure averages measured either 7 times or 5 times during the nictemeral rhythm In a small amount of cases, 4 eyes in the normal patients group and 3 eyes in the glaucomatous patients group, important data are excluded by the 5 values nictemeral curve. CONCLUSION: We believe that a 5 determinations pressure curve is sufficient enough to estimate correctly a glaucomatous patient regarding his intraocular pressure regimen. PMID- 18389786 TI - [Cervical metastatic adenopathy from the oropharynx cancer]. AB - Cervical metastatic adenopathy represents quite often the first symptom in many cancers of the upper respiratory and digestive pathways and might be the first symptom in 20-25% of the oropharyngeal cancers. MATERIAL AND METHOD: The study is based on 243 cases of oropharyngeal cancer admitted to the ENT Clinic, "Sf. Spiridon" Hospital Iasi within the last ten years. RESULTS: By analyzing the evolution of 243 cases of oropharyngeal cancer we conclude: because of the lack of symptoms, the alarming symptom is very often the cervical adenopathy so that the diagnosis is established in the late stages. On the other side, the abundance of the lymphatics of the pharynx make possible the metastatic adenopathy to show in the early stages of evolution of an oropharyngeal cancer. CONCLUSION: Radical neck dissection followed by radiotherapy is proved to be so far, the optimal treatment for cervical metastatic adenopathy. PMID- 18389787 TI - [Early contrast-enhanced MR imaging assessment of femoral head viability after femoral neck fracture]. AB - AIMS: To evaluate the use of MR imaging, before and after i.v. administration of Magnevist, for assessing the femoral head perfusion after femoral neck fracture. Evaluation of femoral head viability is important because the outcome of internal fixation is adversely affected by the development of capital osteonecrosis. METHODS: We performed MRI of the femoral head in 48 hours of injury, on 10 patients with femoral neck fracture. Five patients underwent MR imaging of the hip utilizing fat-suppressed (STIR) sequences and the others, T1-weighted spin echo sequences before and after i.v. contrast administration. MR findings were correlated with radiographic follow-up for at least 12 months. RESULTS: Radiographic follow-up showed femoral head osteonecrosis in two patients. On contrast-enhanced MR images of these patients, the femoral head did not enhance and was lower in signal intensity than were the enhancing femoral shaft and neck distal to the fracture and the enhancing femoral head on the unaffected side. In the patients with persistent perfusion, contrast-enhanced MR images showed a uniform increase in signal intensity in the femoral shaft and neck as well as the femoral head; the femoral head on the fractured side showed contrast enhancement similar to that on the healthy side. CONCLUSIONS: These preliminary results indicate that contrast-enhanced MR imaging may be useful for noninvasive evaluation of femoral head perfusion after femoral neck fracture. MR findings also may aid the clinician in deciding between joint-preserving therapy and hip arthroplasty. PMID- 18389788 TI - [Hemostatic hysterectomy in modern obstetrics]. AB - Haemostatic hysterectomies have been performed for the first time in the XIXth century to reduce the increased maternal mortality and associated morbidity. In Romania the main cause of death through direct obstetrical risk (DOR) is the haemorrhagic syndrome, with a value of 45% of the total number of deaths in the last 15 -16 years, a lot higher then the world average of 25% as evaluated by OMS. Among the deaths due to hemorrhagic syndrome 43% is held by antepartum haemorrhage (placenta praevia, utero-placental apoplexy) and 42% by postpartum haemorrhage. RESULTS: In the past 15 years (1990 - 2005) there have been studied 53,870 births, out of which only 60 have had haemostatic hysterectomy, representing 0.11% of the total number of births. The prevalence of haemostatic hysterectomy is only 1 in every 883 births as a result of surgical teams' efforts to preserve the uterus. The placental pathology (30%) is responsible for most of haemostatic hysterectomy indications. Out of 60 cases under study, none ended with maternal death, while 11 cases ended with fetal death (intrapartum or postpartum). CONCLUSIONS: Morbidity and maternal death can be reduced by effective prenatal care, by identifying high risk patients and by the possibility of caesarean section. Haemostatic hysterectomy remains, in essence, a life saving procedure! PMID- 18389789 TI - [Internal hernia--a rare cause of intestinal obstruction]. AB - Internal hernia is rare its frequency ranging between 0.6 and 5.8%. It results from the protrusion of one or more abdominal viscera (usually small bowel) through an intraperitoneal opening. The opening can be normal (e.g. Winslow foramen), congenital (paraduodenal fossa, ileocecal fossa), or abnormal anatomical entities (after trauma or surgery). The clinical diagnosis of internal hernia is difficult because of the lack of specific signs and symptoms. There is a 63.6% lifetime risk of strangulation and bowel ischemia. In such cases, computed tomography is essential in the preoperative diagnosis because of the high mortality rate (20%) (which justifies its costs). PMID- 18389790 TI - [Ovarian cyst and pregnancy. Conservative management and consecutive emergency cystectomy. Case report]. AB - The association of an ovarian cyst with pregnancy is relatively rare; it may result in serious maternal and fetal complications and its treatment is still controversial. We present a case in which the rarity is given by the association of an 11cm--serous right ovarian cyst (with signs of torsion) with a monofetal 6 gestation week--pregnancy, by the initial conservative management and by the subsequent laparotomy and cystectomy at 16 gestational weeks, required by the acute abdominal pain caused by the torted cyst. The approach was initially conservative followed by laparotomy and cystectomy at 16 gestation weeks, required by the acute abdominal pain caused by the torsive cyst. Pregnancy was, afterwards, uneventful and the patient delivered vaginally, at term, a 3200 g healthy girl. Management options of such a case are discussed. PMID- 18389791 TI - [Zinc involvements in the brain]. AB - Central nervous system (CNS) concentrates almost 10% of total zinc in the human body. Imbalances in zinc concentration are associated with numerous CNS diseases. Zinc deficiency is associated with nervous anorexia, major depression, cognitive impairment, and uncontrolled behavior. Our data reveal that plasma zinc concentration is decreased in major depression and it significantly increases following sertraline or amitriptyline treatment. Also, we found that ZnCl2 administration while inducing morphine-dependence in rats significantly decreases the symptoms of opioid-withdrawal syndrome. Recent data incriminate zinc deficit in the development of encephalopathy following severe impairment of hepatic function. On the other hand, zinc content of certain brain areas in Alzheimer disease is twice that in controls. Parkinson disease is also associated with higher zinc concentrations in the brain than normal. The ratio plasma zinc- other bivalent cations is also important for normal brain function. PMID- 18389792 TI - [Interactions between angiotensin II and theophylline on isolated rat bronchi]. AB - For more than half of century physicians are using theophylline for the treatment of obstructive pulmonary diseases. Because our previously results suggested the amplification of intrapulmonary renin angiotensin system (RAS) on ovalbumin (OVA) induced airway hyperresponsiveness we studied the interaction between theophylline and angiotensin II (Ang II) on normal versus sensitized rats. MATERIALS AND METHODS: we used main left bronchial rings mounted in wire myograph to assess the effects of Ang II and theophylline on airway smooth muscle. RESULTS: On both normal and OVA sensitized rats theophylline did not significantly modify either Ang II contractile effects or Ang II amplification of acetylcholine (ACh)-induced bronchoconstriction. On the other hand, on sensitized rat after antigen challenge, theophylline pretreatment reduced Ang II inhibition of terbutaline--induced relaxation of bronchial rings precontracted with ACh, increasing both EC50 and E(max) of terbutaline effects with 22.04 +/- 3.48% and 19.48 +/- 1.67%, respectively. CONCLUSION: These findings suggested that, in addition to bronchodilatory and antiinflammatory actions, theophylline could block some effects of intrapulmonary RAS activated in pathologically states as antigen sensitization and challenge. PMID- 18389793 TI - [FISH technique in aneuplodies prenatal diagnosis]. AB - Chromosomal disorders are severe and affect 0.9% of the newborns. In these conditions, prenatal diagnosis should be compulsory in every public medical system. MATERIAL AND METHOD: Our study is a retrospective analysis of pregnant women investigated by amniocentesis and FISH technique. RESULTS: We analyzed 233 samples collected between 2004 and 2007 at Iasi "Cuza-Voda" Obstetrics and Gynecology Hospital. The majority of cases were investigated between 16 and 24 weeks of pregnancy. Thirty-eight abnormal cases (16.30%) were identified: 17 cases with 21 trisomy, 16 cases with 18 trisomy, 3 cases with X monosomy, and 2 cases with 13 trisomy. The main reasons for amniocentesis were: advanced maternal age (12.5% abnormal cases) ultrasound abnormalities (26.15% abnormal cases), and biochemical abnormalities (7.14% abnormal cases). CONCLUSION: Our data are in agreement with other studies, and support our results. PMID- 18389794 TI - Benign gastric tumors. Anatomopathological aspects of the gastric wall. AB - Benign tumours of the stomach are quite rare and are discovered accidentally during routine endoscopy or necroscopy. They represent less than 20 per cent of gastric tumours, and their clinical picture consists in pain, bleeding and antropyloric stenosis. MATERIAL AND METHOD: The cases were studied retrospectively over a 10-year period (1995 - 2005) and consist of patients that underwent needle biopsies and surgery in the Timis County Hospital. RESULTS: Benign tumoural pathology was present in 73 cases, 43 (58.9%) in women, 30 (42.1%) in men. The age of the patients ranged between 36-88 years in women, and 31-87 years in men. The material for study consisted in gastric resection pieces and specimens of needle biopsy. In order to establish the histopathological diagnosis and to define the specific type of the damage, the first specimens were stained using morphological methods. Routine morphological investigation was carried out through Hematoxylin-Eosine staining, which was the standard technique used for all the cases. PMID- 18389795 TI - [MLPA technique--principles and use in practice]. AB - MLPA (Multiplex Ligation-dependent Probe Amplification) is a recently introduced method, based on PCR principle, useful for the detection of different genetic abnormalities (aneuploidies, gene deletions/duplications, subtelomeric rearrangements, methylation status etc). The technique is simple, reliable and cheap. We present this method to discuss its importance for a modern genetic service and to underline its multiple advantages. PMID- 18389796 TI - [Hospital management--performance measuring models]. AB - High performance of a hospital follows aims: reaction and needs of the community, integrity of health system, availability of hospital services for all population without difference of social, culture, religion or economic status. This paper reviews some experience in developing countries for understanding hospital performance and methods for measuring the quality of hospital management. All these methods are studied by manager and the staff of Hospital "Sf. Spiridon" Iasi, Romania for a strong medical and economic management. PMID- 18389797 TI - [Contributions to the knowledge of clinical and epidemiological features of essential arterial hypertension in Moldavia, Romania]. AB - Multi-factorial etiology of essential arterial hypertension (EAHT) can benefit of new data by studies which allow a better knowledge of risk factors and their involvement in the disease pathogenesis. Increase in the prevalence values of EAHT and, as a consequence, of its grave complications, represents the major objectives of prevention programmes. MATERIAL AND METHOD: Authors present a clinical epidemiological study on a sample of 932 patients, 441 men and 491 women, aged from 18 to over 65; 876 from urban area and 56 from rural. RESULTS: Familial antecedents with risk for AHT were assessed: EAHT (36.7%), ischemic cardiopathy (13.3%), myocardial infarct (13.7%), stroke (15%), diabetes mellitus (13.2%), obesity (27.5%), and dyslipidemia (31.8%), as well as body mass index (BMI)--34.9% overweight persons and 32.9% suffering from obesity; repartition on blood groups, behaviour type (A--67.8%), sedentary lifestyle (35.8%), stress (extra-familial--32.1%), active tobacco smoking (52.1%), excess coffee (> 3 cup daily--35.7%), alcohol (10.1%) and salt (31.0%) consumption. Paraclinical investigations included: blood pressure measurement, ECG, and echocardiography. Laboratory results showed hyperglycemia in 17.8% of cases, hypercholesterolemia- 70.0% (hypo-HDL--54.3%, hyper-LDL--63.0%), and hypertriglyceridemia in 70.7%. PMID- 18389798 TI - [Assessments of retro-prospective epidemiologic research results regarding morbidity with viral hepatitis A in North-Eastern Moldavia, Romania, between 1970 2006]. AB - Viral hepatitis A (VHA) maintains the first place among hepatitis with various etiologies, by high levels of morbidity, affecting mainly children and teenagers age groups and, sometimes, with major risks in adults and when over- and co infected with other hepatitis viruses. It also is involved in travellers' and nosocomial diseases. MATERIAL AND METHOD: The assessment of VHA morbidity data highlighted that VHA registered yearly values between 106.8 and 307.1% hundred thousand inhabitants, both at the national and Suceava County levels, during the period 1970-1995. RESULTS: An important decline (24.8-47.8% hundred thousand) was registered, between 1996 and 2006. In the Suceava County, after the epidemic peaks from '80-'90 decade, with morbidity of 379.1% hundred thousand in 1989 and 331.8% hundred thousand in 1990, inter-epidemic and epidemic periods with reduced amplitude were registered. In 2006, morbidity reached a low value of 15.0% hundred thousand. CONCLUSION: Our study of "comparative epidemiology", on reported cases and those confirmed by hospitalisation, in the Suceava County, highlighted that the later proportion was under 50%, with an increase during 2005 (> 90%). A significant decrease in multi-annual trend of VHA morbidity registered for reported cases but not for those hospitalised. PMID- 18389799 TI - Maternal death and orphans in Cluj County. AB - The communist government that led Romania before the 1989 Revolution practiced a demographic policy which claimed to protect the family and encouraged women to have minimum five children. MATERIAL AND METHODS: The complex implications of maternal death rate, as a negative demographic phenomenon, were investigated through a descriptive, observational retrospective study made in the Cluj County, Romania, between 1970 and 2004. The study covers all deceased mothers (n = 263 cases) recorded in Cluj County from 1970 to 2004. The data were collected from the primary evidence documents of the Obstetrics-Gynecology Clinics and were analyzed on socio-demographic variables, causes and consequences of death before and after 1990. RESULTS: Up to 1989, the majority of maternal deaths were caused by abortion and its complications. After the changes in the demographic policy in 1990, this cause of maternal death decreased dramatically, the first cause now being obstetrical risk. The age group most affected by mother death was married women between 20-39 years old, coming from the country side and with a low level of education. These deaths left 520 children orphan. CONCLUSIONS: The death of these mothers generated a considerable number of orphans, who were likely to become socially dependent. Since orphans have equal rights to life and social integration with other children, the society should become actively engaged in their upbringing, supervision and special education. PMID- 18389800 TI - [Assessment of the impact of psycho-social environment on toddlers' physical and neuropsychic development]. AB - Psycho-social environment is influencing directly toddler's development, with a major impact in the first three years of life. Family is the first social environment for the infant, and later for toddler, and depending to the quality of this relation will evolve the adult. OBJECTIVE: To assess the impact of psycho social environment on the toddler's development. MATERIAL AND METHOD: We have analyzed by using a questionnaire a number of 200 families, in order to assess their level of environmental risk and to correlate it with the toddlers' development. RESULTS AND DISCUSSIONS: The highest risk scores were obtained by families who sent the toddler in a foster home, followed by the families who use a day care center, and at last by the families where toddler remains in the family's home till he is 3 years old. 51.66% of the toddlers who goes in a day care center and 85% of the toddlers who are sent to foster home are exposed to a relative risk lower by 2.7 times to be retarded compared to the situation of remaining in the natural home. On the other hand, in family environment with a mild risk there are significant more disharmonic children than in the families with a lower risk score (p = 0.0452). The toddler's chances to evolve, by physical perspective, negatively in institutional environment are significantly higher if he comes from a family with mild risk score than in a family with a lower risk score (p = 0.0157). CONCLUSIONS: In every life environment where lives a toddler, confronted with specific problems, should take immediate action the general practitioner, the child development specialist and the environmental health expert, in a coordinate manner, to identify and correct all the aspects who may disturb the normal evolution of the toddler's growth and development. PMID- 18389801 TI - [In vitro activity of ertapenem against staphylococci]. AB - Ertapenem is a new carbapenem with an excellent activity against different Gram negative and Gram-positive bacteria. Its good antibacterial activity is closely related to high hydrolysis resistance by bacterial beta-lactamases. MATERIAL AND METHOD: We evaluated antimicrobial activity of ertapenem by comparison that of methicillin, gatifloxacin, cefepime and linezolid against 455 staphylococci collected during 2006-2007 period from healthy individuals. RESULTS: Minimum inhibitory concentrations were determined by agar dilution technique. Ertapenem has an activity comparable to the other tested agents. Resistance rate was 1% for all investigated strains. PMID- 18389802 TI - [Perio Type--the premises of a successful therapy]. AB - The Perio Type implants represent a current alternative for treatment for the implant-prosthetic rehabilitation, the design and the typological variety leading to a pertinent selection dictated by the particularity of the clinical case. The processed element method is eloquent for the biomechanical behavior of these types individualized implants on the clinical archetypes, crystals that have forwarded the clinical applications. The most often used implants from this category have been those from the x-class, characterized by a high degree of perio-integration. PMID- 18389803 TI - [Functional aspects of lower jaw dynamics--EMG study]. AB - The paper presents a study of muscular forces using EMG records for temporal and masseter muscles on different occlusion situations. Each muscle activity was investigated related to different occlusion situation in order to be able to determine correlations with possible diagnostic and treatment of dental system malfunctions. The results regarding the practical determination of symmetrical indexes and muscle activity indexes were compared with specific references. PMID- 18389805 TI - [CT exam and 2D and 3D reconstructions in facial tumors diagnosis]. AB - This study presents the CT imaging of a group of maxillofacial tumors that include benign tumors and primary and secondary malignant processes. MATERIAL AND METHOD: All patients were CT explored with and without intravenous contrast, using specific head and neck protocols. RESULTS: We studied the topography of the tumors, their impact on neighborhood regions and post-treatment results. The diagnostic of all patients had histological confirmation. In maxillofacial tumors and tumor-like diseases, 3D imaging was significantly superior to 2D axial imaging in showing the tumor shape and spatial relationships between tumors and surrounding structures. Two dimensional imaging and MPR imaging were excellent to reveal internal structure and pathological changes of tumors. 2D imaging and MPR imaging also achieved better results in showing tumors extended to soft tissues. PMID- 18389804 TI - [Considerations regarding a prediction method for third mandibular molar eruption]. AB - Third mandibular molar eruption is an unpredictable event. MATERIAL AND METHOD: We have studied 250 patients who required orthodontic treatment in the Department of Orthodontics and Dentofacial Orthopedics, University of Medicine and Pharmacy "Gr.T. Popa" Iasi. Orthopantomograms were traced for all patients at the beginning of the orthodontic treatment. Some patients presented first lower molar removal before orthodontic treatment. RESULTS: We evaluated the difference between posterior available space and third lower molar mesiodistal dimension in order to establish the predictable values for third molar eruption. Our results show that only 17% of lower third molars have 100% chances for eruption in non extraction sample, comparative with 65-70% in extraction group. 38% of third mandibular molars have a poor prognosis for eruption in non extraction group and only 14% for the pretreatment extraction group. PMID- 18389806 TI - Effects of Ocimum basilicum L. extract on experimental acute inflammation. AB - Our study investigated the effects of Ocimum basilicum L. tincture (1:10) in acute inflammation induced with turpentine oil (i.m. 0.6 ml/100 g b.w.) in Wistar male rats. MATERIAL AND METHODS: The results were compared with those from a positive control group with experimental inflammation and a group treated with diclofenac (30 mg/100 g b.w.). The effects were assessed by measuring total leukocyte count and differential leukocyte count expressed as a percentage, a test of in vitro phagocytosis, and the evaluation of nitric oxide synthesis by measuring the metabolites, nitrites and nitrates, and the co-product citrulline. RESULTS: Ocimum basilicum tincture significantly reduced the total leukocyte count, monocyte percentage, activation of circulating phagocytes, but had a slight inhibitory effect on NO synthesis. Compared to diclofenac, Ocimum basilicum tincture had a smaller inhibitory effect on all tested parameters. CONCLUSION: The tested Ocimum basilicum tincture has important anti-inflammatory effects on bone marrow acute phase response and a reduced one on NO synthesis. PMID- 18389807 TI - Polysaccharides fraction isolated from ginkgo biloba folium: immunopharmacological properties. AB - Many macromolecular compounds of vegetal polysaccharides type develop important immunostimulatory properties. MATERIALS AND METHODS: Crude macromolecular fraction was isolated from the leaves of Ginkgo biloba L. (codified as PGfol) harvested from Botanical Gardens of Iasi, Romania. The hot water extract was partially purified and submitted to chemical and immunopharmacological studies. The immunostimulatory activity of PGfol was further investigated by a series of tests on rats: phagocytosis capacity of PMN cells by NBT test, serum complement activity and determination of the activity of splenic T-lymphocytes. RESULTS: PGfol has a good immunostimulatory activity which is equal or higher to Levamisole (positive control). PMID- 18389808 TI - [Physico-chemical characterisation of Zn-flavonols complexes]. AB - The aim of flavonols complexation is to improve their specific activity. MATERIAL AND METHOD: The flavonols sources were Morus alba L. and Morus nigra L., zinc being the biological active ion. Physico-chemical characterization was made by thin-layer chromatography, UV-VIS spectrophotometry, IR spectrometry and polarography. RESULTS: The complexes results were within acceptability limits, but their obtaining is economically inefficient. PMID- 18389809 TI - [Study on an injectable biomimetic hydroxyapatite--atelocollagen composite]. AB - The paper presents a procedure for preparing mineralized collagen nanofibrils, mixed with deficient nanoparticulate calcium hydroxyapatite in a ratio close to that found in living bone structures. Such a composition might represent an injectable osteoconductive constituent, used in surgical kits for bone reconstruction. The FT-IR spectra of the prepared composite reveals a weak absorption at 871.82 cm(-1), indicating the nucleation of hydroxy-apatite at the acidic atelocollagen functions. More than that, a 45 cm(-1) ipsochromic shift of carboxylic band suggest a strong ionic interaction with hydroxyapatite. The X-Ray spectra show a high resemblance between the synthetic composite and the Fluka hydroxyapatite, but only for 2theta angles higher than 30 degrees. Below that value the protein presence induce crystalline disorders. PMID- 18389810 TI - [Optimisation of hip endoprosthesis election by using finite element analysis]. AB - The main aim of this study is to present an optimized method for election of hip endoprosthesis by using finite element analysis. MATERIAL AND METHODS: Four types of hip endoprosthesis with different design and fixation type were considered for this study. First step is three-dimensional reconstruction to one femur using CT sections; the second represent the hip endoprostheses components 3D reconstruction, virtual implantation of each endoprosthesis in femur, then finite element analysis. RESULTS: The calculated Von Mises stress as a result of finite element analysis indicates the optimum endoprosthesis by considering the stress on endoprosthetic nail and bone. PMID- 18389811 TI - [Selective separation of cinnamic and p-methoxycinnamic acids. The study on reactive extraction from mixture]. AB - Cinnamic and p-methoxycinnamic acids have been separated from their mixture by reactive extraction with Amberlite LA-2 dissolved in n-heptane. The efficiency and the selectivity of their separation is controlled by the pH-value and extractant concentration, the optimum conditions being the pH-value of aqueous phase of 2 and the extractant concentration into the organic phase of 10 g/l. PMID- 18389812 TI - [Scientific collaboration of the Society of Medicine and Natural Science in Jassy with prominent European scientists during the first decades of its existence]. AB - This article details the scientific collaboration of the Society of Medicine and Natural Science in Jassy with prominent European scientists during the first decades of its existence. The intensity of the scientific contacts of the Society of Medicine and Natural Science in Jassy arise from detailed analysis of the correspondence that outlasts time in the state archive of Jassy. 75% of this correspondence was written in German, and most of it was sent from the German Confederate or the Austro-Hungarian Monarchy. This influence and contribution of German science in Moldavian natural science development, was undoubtedly in the first half of the 19th century. This can be attributed to dr. Iacob Cihac, one of the founders of the Society, who was born in Aschaffenburg (Germany) and studied medicine in Heidelberg (Germany), before he moved to Moldavia. Based on the initiative of drs. Cihac and Zotta, and not least the financial support of a part of the Moldavian high class and the Moldavian government, the Society of Medicine and Natural Science in Jassy was founded in 1833. This became the first scientific society in the territory of modern Romania. Since the inception of the Moldavian Society of Medicine and Natural Science in Jassy, it has pushed the boundaries of a simple scientific society. This society provides an encyclopedic framework of most of the scientific subjects of the 19th century (medicine, pharmacy, natural science, agronomy, paleontology and geology). It played a major role during the democratization of the Moldavian education system, for example by founding a medical school teaching in the Romanian language in Jassy. The society survived and continued to maintain scientific activities during all the political changes in Moldavia during the 19th century, particularly the revolution of 1848 and the unification process of Romania. The influence and activity of the society in Jassy has continued to make a significant contribution to science and education development in Romania, until today. PMID- 18389813 TI - The riches of e-prescribing. Insurers will say, do pretty much anything to get physicians to prescribe medications electronically. Ever wonder why? AB - Insurers are leading the charge for e-prescribing, a technology that promises to save them big bucks. But they don't always agree on the best way to ensure adoption. WellPoint is sticking with a sponsorship-based model. "Unless you have the health plans pushing this, it will take a mandate from Congress or decades for e-prescribing to be adopted widely," says WellPoint's Charles Kennedy, left. PMID- 18389814 TI - Endorsing e-prescribing. FAH's Kahn says it helps reduce errors. PMID- 18389815 TI - Tying nurse quality to payment. CMS may link reporting on RN indicators to hikes. PMID- 18389816 TI - Cuomo's rant on rates. N.Y. attorney general to sue Ingenix, UnitedHealth. PMID- 18389817 TI - Stop blaming the patient. Penalizing workers for personal health decisions is off the-mark. PMID- 18389818 TI - Stop those rebate checks! To stimulate the economy, invest in healthcare, where the jobs are. PMID- 18389820 TI - Largest military hospitals. Ranked by number of total beds. PMID- 18389819 TI - The hope of HIPAA 2. Data stewardship must replace minimal compliance. PMID- 18389821 TI - How can case managers incorporate health promotion into their practices? PMID- 18389822 TI - Weight gain among women during smoking cessation: testing the effects of a multifaceted program. AB - Cigarette smoking continues to be the leading preventable cause of chronic illness in the United States. A high percentage of smokers attempt to quit each year; however, smoking cessation success rates are low. Adding an exercise program to traditional therapies can increase smoking cessation rates among women. An experimental design was used to test two hypotheses. The first was that women who engage in a multifaceted approach to smoking cessation that includes a structured exercise program, nicotine replacement therapy (NRT), and weekly counseling sessions will experience less weight gain during an 8-week smoking cessation program and at the 4-month follow-up session than women who engage in NRT and weekly counseling sessions alone. The second was that women who engage in a structured exercise program with NRT and weekly counseling sessions will have higher smoking cessation rates at the end of the 8-week smoking cessation program and at the 4-month follow-up session than women who engage in NRT and weekly counseling sessions alone. PMID- 18389823 TI - A measure of university employees' exposure to risk factors for work-related musculoskeletal disorders. AB - This study examined the computer workstations of 50 faculty and staff at a regional comprehensive university in the southeastern United States. Data were collected using a subjective questionnaire and the observational Computer Workstation Analysis tool. The researchers found no significant differences between participants' perceptions of their risk for work-related musculoskeletal disorders (WRMDs) and observations of workstations at the worksite. The researchers recommend developing an ergonomics program delivered by occupational health team members with knowledge and skill in WRMD prevention, identification, and treatment. PMID- 18389824 TI - Addressing environmental health Implications of mold exposure after major flooding. AB - Extensive water damage resulting from major flooding is often associated with mold growth if materials are not quickly and thoroughly dried. Exposure to fungal contamination can lead to several infectious and noninfectious health effects impacting the respiratory system, skin, and eyes. Adverse health effects can be categorized as infections, allergic or hypersensitivity reactions, or toxic irritant reactions. Workers and building occupants can minimize their exposure to mold by avoiding areas with excessive mold growth, using personal protective equipment, and implementing environmental controls. Occupational health professionals should encourage workers to seek health care if they experience any symptoms that may be linked to mold exposure. PMID- 18389825 TI - American Nurses Association's best practices in seasonal influenza immunization campaign. AB - The American Nurses Association (ANA) is concerned about the unacceptably low rates of seasonal influenza vaccination among registered nurses and other health care personnel. Despite well-documented evidence of the importance and benefits of influenza immunization of health care personnel, seasonal influenza vaccination rates remain below 50%. In response, ANA developed the 2006-2007 Best Practices in Seasonal Influenza Immunization Campaign to assist health care organizations in conducting successful seasonal influenza immunization programs. Best practices in the top recognized programs have been compiled to represent the successful elements in seasonal influenza vaccination programs that result in increased vaccination acceptance and accessibility among health care professionals. ANA developed a brochure, articles, and other tools reflecting the best practices to assist other organizations nationwide in planning their seasonal influenza immunization programs. PMID- 18389826 TI - Alleged solvent exposure and deviation from procedures. PMID- 18389827 TI - Navigating the OSHA website. PMID- 18389828 TI - Escaping from the silo? PMID- 18389829 TI - Exostectomy for chronic midfoot plantar ulcer in Charcot deformity. AB - Charcot midfoot ulcers are rare and very difficult to heal, with surgery being an option. This retrospective study assessed healing rates, complications, and the incidence of re-ulceration and other foot ulcer problems following exostectomies PMID- 18389830 TI - Use of autologous platelet-rich fibrin on hard-to-heal wounds. AB - This retrospective study found that use of autologous platelet-rich fibrin on a range of hard-to-heal wounds achieved full healing or a significant reduction in wound diameter with no adverse effects. Prospective studies are now needed PMID- 18389831 TI - Use of an ionic sheet hydrogel dressing on fungating wounds: two case studies. AB - Two patients with terminal cancer presented with painful, highly exuding and intensely malodorous wounds. In both cases, use of an ionic hydrogel controlled the malodour, absorbed the exudate and reduced pain, improving quality of life. PMID- 18389832 TI - An economic evaluation of VAC therapy compared with wound dressings in the treatment of diabetic foot ulcers. AB - OBJECTIVE: To determine the cost-effectiveness ofVacuum Assisted Closure (VAC) therapy (KCI Medical), based on a comparison with both traditional and advanced wound dressings, for the treatment of diabetic foot ulcers in the US. METHOD: A Markov model was designed to estimate the cost per amputation avoided and the cost per quality-adjusted life year (QALY) of VAC therapy, compared with both traditional and advanced dressings. Over a one-year period the Markov model simulated 1000 patients using transition probabilities obtained from the literature. The health states used in the model were: uninfected ulcer; infected ulcer; infected ulcer post-amputation; healed; healed post-amputation; amputation; and death. Patients initially treated with VAC switched to the advanced dressing after three months of treatment if their wound remained unhealed. Patients treated with traditional or advanced dressings were assumed to continue with their treatment for the full 12 months if they remained unhealed. RESULTS: The model results demonstrate improved healing rates (61% versus 59%), more QALYs (0.54 versus 0.53) and an overall lower cost of care ($52,830 versus $61,757 per person) for patients treated with VAC therapy compared with advanced dressings. Vacuum Assisted Closure was also shown to be a dominant intervention when compared with traditional dressings. CONCLUSION: The model results indicate thatVAC therapy is less costly and more effective than both traditional and advanced dressings. The results are robust to changes in key parameters, including the transition probabilities, the cost ofVAC therapy and the utility weights applied to health states. PMID- 18389833 TI - Topical negative pressure therapy for para-ileostomal ulceration in a patient with Behcet's disease. AB - BehCet's disease is a chronic, relapsing, inflammatory disorder. This case report describes how use of an improvised topical negative pressure (TNP) device in a patient with a non-healing para-ileostomal ulcer with Behcet's disease. PMID- 18389834 TI - A question of rigour. PMID- 18389835 TI - Stuck on the couch. PMID- 18389836 TI - Wanted: someone to play god. Science has given us childbirth miracles. Now we need laws to create some boundaries. PMID- 18389837 TI - Leadership-organizational culture relationship in nursing units of acute care hospitals. AB - The phenomena of leadership and organizational culture (OC) has been defined as the driving forces in the success or failure of an organization. Today, nurse managers must demonstrate leadership behaviors or styles that are appropriate for the constantly changing, complex, and turbulent health care delivery system. In this study, researchers explored the relationship between nurse managers' leadership styles and OC of nursing units within an acute care hospital that had achieved excellent organizational performance as demonstrated by a consistent increase in patient satisfaction ratings. The data from this study support that transformational and transactional contingent reward leaderships as nurse manager leadership styles that are associated with nursing unit OC that have the ability to balance the dynamics of flexibility and stability within their nursing units and are essential for maintaining organizational effectiveness. It is essential for first-line nursing leaders to acquire knowledge and skills on organizational cultural competence. PMID- 18389838 TI - The AACN Synergy Model for Patient Care: a nursing model as a Force of Magnetism. AB - One of the 14 Forces of Magnetism requires a health care organization to have a professional model of care. The eligibility requirements stipulate that this model must be utilized throughout the health care system and that the same philosophy must be used throughout the system. The American Association of Critical-Care Nurses Synergy Model for Patient Care describes nursing practice based on eight patient characteristics, and also describes eight nurse competencies. The core concept of the model is that the needs or characteristics of patients and families influence and drive the characteristics or competencies of nurses. Synergy results when the needs and characteristics of a patient, clinical unit, or system are matched with a nurse's competencies. The synergy model is an excellent framework to organize the work of patient care throughout the health care system. It can be used and applied in various ways and provides a comprehensive framework for assuring success in building a philosophy that supports the Forces of Magnetism. PMID- 18389839 TI - Building a nursing research fellowship in a community hospital. AB - Often, registered nurses feel that they are using "evidence" to guide practice, but in reality the sources of evidence are not based on research. The nursing leadership team at Northwest Community Hospital was committed to integrating evidence-based practice and nursing research into the clinical setting for RNs at all levels of the organization. The scope of the fellowship program enabled direct care RNs to receive protected release time for a 12-month period to develop a research proposal, implement the study, and disseminate the findings. The Nursing Research Fellowship Program was successful from the perspective of the Magnet appraisers who recognized the program as a Magnet exemplar, the chief executive officer, the CNE, members of the nursing leadership team, professional colleagues, the fellows and partners, and registered nurses at all levels in the organization. Nursing research is now part of the professional nursing culture. PMID- 18389840 TI - The nursing shortage continues as faculty shortage grows. AB - To combat the nursing shortage, efforts to promote nursing as a career have been successful. However, academic nursing institutions are not adequately prepared for this new influx of applicants. The lack of faculty to educate the growing demand for baccalaureate-prepared RNs directly impacts the nursing shortage. The nursing shortage thus directly impacts safe patient care. The main reasons for the lack of faculty to meet the demand for more nurses include the increased age of the current faculty and the declining number of years left to teach, expected increases in faculty retirements, less compensation for academic teaching than positions in clinical areas for master's-prepared nurses, and finally, not enough master's and doctoral-prepared nurses to fill the needed nurse educator positions It is in the best interest of the nursing profession to do what it does best by incorporating the nursing process to solve the faculty shortage and secure its future in order to protect the lives of patients. PMID- 18389841 TI - Cost-effective care a phone call away: a nurse-managed telephonic program for patients with chronic heart failure. AB - Health care providers and health care systems are challenged to find cost effective ways to address the costs associated with heart failure. A multidisciplinary team of nurses, physicians, pharmacists, and dieticians was assembled at New Hanover Regional Medical Center (Wilmington, NC) to develop strategies to decrease the readmission rate without compromising patient care. The team developed a disease management program that included comprehensive inpatient education, as well as an outpatient telephonic program to reinforce education after discharge. Goals were to reduce readmissions of patients with heart failure, to decrease the cost per case of each patient with chronic heart failure, and to reduce the length of stay for patients who were readmitted. The CHF Telephonic Program was extremely successful in meeting patient-focused and organizational goals related to readmissions, length of stay, and cost of care. PMID- 18389842 TI - Workload measurement in a community care program. AB - The delivery of services in the community is a expanding area of health care due to the rapid shift from institutional to community care. However, limited attention has been given to accurately predict or measure nursing resource utilization. A workload measurement tool developed by the community care program at WestView Health Centre (Stony Plain, Alberta, Canada) sought to address the unique challenges of determining workload while caring for clients in their own homes. The implementation of this tool facilitated improved understanding of work requirements associated with each client on caseload. The continued use of this tool will lead to refinement of case mix and professional utilization, and may show benefits in the future for novice nurse growth, staff retention, and satisfaction. PMID- 18389843 TI - Envisioning and implementing new directions for health care. AB - Across the entire health sector, federal analysts indicate that in 2006 approximately $2.1 trillion were spent on health care, over $7000 per person. With expenditures at this level, it is no surprise that people are asking whether the public receives value commensurate with the financial investment. While strides are being made on a number of quality of care fronts, given the financial resources being consumed, overall forward momentum is glacial. Educating the public is a particular challenge given amplified sound bites and attack ads designed to preserve the status quo. All of this underscores that nursing has a role not just in implementing new directions in health care but envisioning them as well. PMID- 18389844 TI - Preceptors for non-clinical employees: extending the value. AB - Although preceptors have been utilized for many years to orient nurses, non clinical employees have not had the benefit of having consistent, competent, standardized guidance during their orientation period. A preceptorship program was implemented in Ambulatory Care Services for non-clinical employees to improve the orientation experience and to make new employees feel valued. The preceptor program was designed to standardize the information given and skills taught, to increase competence in all job tasks, to assure completion of orientation documents, to reduce staff turnover and increase job satisfaction, and to reduce orientation costs to the organization. Many improvements have been realized related to these goals, including improved staff satisfaction and reduced turnover. PMID- 18389845 TI - Leadership and learning from the politicians. AB - As we observe, the political candidates, we have a wonderful laboratory to discover what works and what does not in engaging followers to buy into the leader's message. Relationships and the ability of the candidate and the leader to feel a sense of personal attachment are important keys to success. Leaders who create this sense of magnetism often can tell the compelling stories that connect with people in the organization. The successful leader must first make the commitment to practice evidence-based leadership by examining best practices and modeling her/his leadership practice after these models. PMID- 18389846 TI - Using a spreadsheet/table template for economic value added analysis. AB - Translating clinical research into practical applications that are cost effective has received significant attention as staff nurses attempt to expand new knowledge into an already complex daily workflow. spreadsheet/table template created in a word processing format can assist with setting up and carrying out the analysis of costs for comparing different approaches to routine activities. By encouraging nurses to take the initiative to examine parts of everyday nursing practice with an eye to cost analysis, significant contributions can be made to maximizing the bottom line. PMID- 18389847 TI - Next generation workforce. AB - The health care industry has become a very complex business. CQsts are rising and resources such as funding and human capital are diminishing. Human capital resources are about to reach true crisis proportions. The vital workforce we have counted on is expected to begin thinning as large numbers of Boomers retire. Not only does this deplete the workforce from a pure numbers perspective, but it also affects intellectual capital and institutional memory. Generational trends and characteristics have affected the workforce environment and will continue to do so as another generation continues to enter the workforce. Generation Y, also tagged Nexter, offers core values that can bring positive changes to the health care workforce. Technology continues to change at lightning speed. Embracing new technology and using it to refine the way we do business will help deliver success. Meaningful strategic plans are needed to change the model of business delivery and employee care in our future workforce. PMID- 18389848 TI - After the tempest. PMID- 18389849 TI - Everything matters to marketers. PMID- 18389850 TI - The new brand landscape. PMID- 18389851 TI - Marketing that pulls. PMID- 18389852 TI - Please stay on the line. PMID- 18389853 TI - Meet your new customers. PMID- 18389854 TI - Targeting or tailoring? PMID- 18389855 TI - Back to the future. PMID- 18389856 TI - 10 steps to reaching physicians. PMID- 18389857 TI - A vote for cultural competency. PMID- 18389858 TI - The real potential in proactive health management. PMID- 18389859 TI - [Postmortem dismemberment/mutilation--medicolegal and criminalistic evaluation of the autopsies performed by the Institute of Legal Medicine at the Hanover Medical School]. AB - Among all the autopsies performed by the Institute of Legal Medicine at the Hanover Medical School between 1978 and 1998. 16 cases of postmortem dismemberment/mutilation were evaluable under criminalistic and medicolegal aspects. In the present study, particular attention was paid to the psychosocial circumstances of the perpetrator's and victim's lives and also to the classification of the different types of postmortem dismemberment/mutilation illustrated by case reports. Apart from an unfavourable personal life situation at the time of the offence, essential predisposing factors were: poor integration in society and family, occupational problems, drug abuse and mental diseases. As the interactions between these factors differ in every single case and may be combined with other factors, the analysis of such homicides should always be based on the assessment of the individual case. PMID- 18389861 TI - [Death after anal "fisting"]. AB - A 45-year-old homeless woman was found dead at her usual sleeping place. Apart from traces of blood on the lower abdomen of the body, the police investigations did not produce any clues pointing to an unnatural death. At autopsy, it was found, however, that death had been caused by extensive disruptions of the intestine. After being confronted with the results, the sexual partner of the victim admitted manual anal penetration, but claimed that this had been done by mutual agreement. The court did not accept that statement and sentenced him to life imprisonment for murder. The frequency of such fatal outcomes of anal penetration, the relationship between the perpetrator and the victim and the special features at the scene are discussed. PMID- 18389860 TI - [Homicide-suicide by a single gunshot to the head]. AB - Apart from typical suicides, there are sometimes unusual deaths from gunshots to the head. The presented case is a double death from the autopsy material of the Institute of Legal Medicine in Berlin. The report deals with a murder-suicide of a man, who killed himself and his 2-year-old daughter by a single gunshot. After having administered a tranquilizer to the child, he held her head to his own and fired a contact shot to his right temple so that the bullet passed through both heads. PMID- 18389862 TI - [Suicidal gunshot to the nape from a small-bore rifle]. AB - A 55-year-old man was found dead on the bed lying on his side in a pool of blood with a bullet entrance hole in the nape. Behind his body, a semi-automatic rifle Remington Nylon, cal. .22 LR, was detected. As the gunshot entrance site was rather unusual for suicide, a forensic autopsy was performed, which showed a contact shot to the nape with the bullet path running upward to the left frontal area. The man had been treated with citalopram for delusional depression, so that a suicidal act seemed plausible, and the autopsy and criminalistic findings were also compatible with this assumption. A remarkable fact of the present case is that a long firearm had been used. Reports on suicidal shots to the nape are comparatively rare in the medicolegal literature and usually refer to pistols or revolvers. PMID- 18389863 TI - [Complex brain malformation as a rare cause of neonatal death]. AB - A dead female neonate was brought to a children's hospital by the mother. The MRI scan suggested a malformation of the brain. Because of the other circumstances of the case (the mother left the hospital unauthorizedly), a forensic autopsy was ordered, in the course of which the brain was removed while the head was totally immersed in water. This method, which was introduced by Prahlow et al., helps to obtain intact brain specimens without interfering with the necessary medicolegal preparations to determine whether the child was born alive. Neuropathological examinations classified the cerebral malformation as (lethal) lissencephaly. Further investigations showed that the mother had lived in Germany without a residence permit. PMID- 18389864 TI - [Unusual suicide by mechanical neck compression]. AB - A 36-year-old man was found dead with his neck being jammed between the hydraulic tail lift and the side wall of a truck. Based on the investigations, the technical facilities of the tail lift, the results of the postmortem examination and the autopsy it could be demonstrated that the victim, who suffered drom depression, had intentionally brought himself into this position and committed suicide by compression of the neck. The pathomorphological findings and the technical reconstruction of this unusual death are presented. PMID- 18389865 TI - [New therapeutic strategies to the advanced non small cell lung cancer]. AB - Lung cancer is an elevated mortality disease in spite of conventional therapy. Today it is possible to identify some molecules, as the epidermal growth factor receptor, involved in the earliest molecular alterations ofcancerogenesis. Here we show the inhibitors of the epidermal growth factor receptor as a therapeutic mean. PMID- 18389866 TI - [Advances in biologic therapy for inflammatory bowel disease]. AB - The medical management of inflammatory bowel disease has considerably changed thanks to the biologic agents coming. In this review a critical evaluation of controlled studies with biologic agents for the management of both Crohn's disease and ulcerative colitis is presented. The efficacy of these agents in moderate to severe ulcerative colitis and Crohn's disease has been one of the most important advances in the past decades. PMID- 18389867 TI - [Parents, children and parenting in cancer patients: a still poorly addressed issue in the global management of neoplastic diseases]. AB - More than 25% of cancer patients in western countries have less than 18 years old children. While increasing attention has been given to various psychosocial issues related to neoplastic disease, the impact of parental cancer on psychosocial 'functioning' of children and adolescents are still poorly explored. Similarly, the role of parenting concerns on quality of life, compliance to the disease and treatments and therapeutic choices are not sufficiently addressed. Usually, cancer patients are reluctant to openly inform their children about their disease. Such "protective" attitude may cause anxiety and psichological distress in children and affect the coping capability of the whole family. Lack of communication may increase the sense of sadness, grief and despair, experienced by children whose parents are ill and induce long-term psychological consequences. Parenthood, on the other side, carries additional concerns to cancer patients which may render disease management more challenging and painful. The oncology team must favour, through appropriate support programs, communication between patients and their children to ensure a better psychological outcome from a stressful situation deeply affecting quality of life of patients and their families. PMID- 18389868 TI - [Cost-effectiveness of oral ibandronate, i.v. zoledronic acid and i.v. generic pamidronate in the management of metastatic bone disease in breast cancer patients: two parallel studies]. AB - Two parallel cost-effectiveness studies comparing oral ibandronate and i.v. zoledronic acid and generic pamidronate have been performed. Two global economic models were adapted to the Italian National Health Service within the management of metastatic osteolysis in breast cancer patients undergoing chemotherapy and hormonal therapy. Total cost was respectively Euro 100 and Euro 418 lower per patient for ibandronate, vs. zoledronic acid and pamidronate in chemotherapy treated patients, and Euro 337 and Euro 419 lower in patients undergoing hormonal therapy; also, with a small increment in QALYs, ibandronate resulted as the dominant therapeutic option. PMID- 18389869 TI - [Usefulness of the follow-up in outpatients affected by liver cirrhosis]. AB - Since 2000 to 2004 we enrolled 80 patients affected by liver cirrhosis: after two year's follow-up in our Outpatients' Department we gained a survival rate of 23%. These data are remarkable because we took care of aged patients (mean age 70 years) with comorbidity (diabetes mellitus, gallstones, hypertension, etc.). PMID- 18389870 TI - [Acute asthma and coma in Emergency Department: a case report]. AB - Acute asthma is a common medical emergency. The goals of therapy are therefore to remove hypoxaemia, improve airflow limitation and prevent death. We describe the case of a 51-year-old woman with asthma admitted to Emergency Department after complaining acute respiratory failure and coma. The improvement of respiratory failure, after administration of high concentration of oxygen, beta-2 agonist and corticosteroids, wasn't associated at improvement of consciousness. The deterioration of blood gases dont justified coma. Having a suspicion the patient takes some benzodiazepines, we administered flumazenil and the patient improved adequately. PMID- 18389871 TI - [Aromatase inhibitors in advanced breast cancer]. AB - Benefits from hormonal therapy in patients with hormon-sensitive breast cancers are widely and well known. A great challenge for clinicians is represented by the possibility to use citoreductive primary hormonal therapy in patients with locally advanced breast carcinoma. Five-year treatment with tamoxifen has been for long time the gold standard in adjuvant settings. In post-menopausal women affected by breast cancer larger than 3 cm and expressing estrogen receptors (ER+), clinical trials have shown that few months treatment with aromatase inhibitors can downsizing tumors. This citoreduction is greater than the one obtained with tamoxifen and it makes conservative surgery possible in most part of cases. PMID- 18389872 TI - [Sydenham's chorea: diagnosis and treatment]. AB - Sydenham's chorea, the neurological manifestation of rheumatic fever, is the most common acquired chorea of childhood. This disorder is secondary to an autoimmune response against basal ganglia in the brain, induced by prior streptococcal infection. Chorea is a major criterion for the diagnosis of rheumatic fever and can also be the presenting feature of the disease. The incidence of rheumatic fever has been markedly decreased by improved socioeconomic conditions and increased antibiotic prescription. In the last two decades, however, this disease has reappeared and should be taken into account in the evaluation of children with movement disorders. In this paper, the author aims to present clinical, diagnostic and therapeutic findings of Sydenham's chorea patients. PMID- 18389873 TI - [Methodology of pharmacoeconomic studies in oncology]. AB - Aim of the paper is to outline the methodology of pharmacoeconomic studies dealing with therapeutical interventions. The specificities of the cost evaluation are described, highlighting that the perspective of the evaluation must be always defined. The main types of pharmacoeconomic analyses are discussed, particularly the cost-effectiveness analysis (CEA) and the cost utility analysis (CUA), and, in both cases, the importance of the incremental cost ratio is stressed. Moreover, sensitivity analysis is introduced as a tool to evaluate the stability of the results, when there is uncertainty in some estimation. Finally, the importance of pharmacoeconomic models is highlighted, although a great caution should be adopted in interpreting their results. PMID- 18389874 TI - [Oncological disease between economic ties and a health humanization policy]. AB - Since 1992 the Italian National Health Service has been characterized by the introduction of managerial concepts. The aim of this article is to analyze the content and impact of the implementation of management and the most significant changes in health care organization and the functioning of the system. The changing organization of health care requires the health care professional to deal with a variety of new demands and, in some cases, to alter his traditional professional practice: the respect of human dignity through empowerment. The general content involves pressures on the health care professional to adapt to the new corporate culture of managed care. At the same time, the expressed psychosocial needs of the cancer patient necessitates increased attention. PMID- 18389875 TI - [Possible exposure to rabies in anamnesis: rabies advice in the Netherlands]. AB - Anamnestic incidences of four patients have highlighted the potential risk ofexposure to rabies. The first patient was a 30-year-old woman who rescued a bat from the mouth of her dog; it bit her on the right wrist. In the Netherlands, bats may be infected with the Lyssa virus. The Preparedness and Response Unit (PRU) of the Centre for Infectious Disease Control (CIDC) advised human rabies immunoglobulin (HRIG) and a full vaccination programme. The second patient was a 37-year-old woman, who caught a 'sick' squirrel and was subsequently bitten on her left hand. The advice was not to use post exposure prophylaxis since rabies is not prevalent amongst squirrels in the Netherlands. The third patient, a 55 year-old man, was bitten on his right calf by a dog in Sri Lanka. He was treated with HRIG and given the full vaccination course. The fourth patient was a 14 month-old boy who was scratched on the face by a cat in Turkey. He immediately received the first vaccination and upon return to the Netherlands was treated with HRIG and the other vaccinations. All patients remained without symptoms. A structured approach for risk assessment of each potential rabies incident is possible. It requires balancing a number of criteria: the species of animal, the endemicity of rabies in a country, the observed health or vaccination status of an animal, whether the animal can be tested for rabies, if the exposure was provoked or unprovoked, the type of injury and its location on the body of the injured, and the time interval between administration of HRIG and vaccine. In the Netherlands all health care providers are expected to perform a proper risk assessment. They may seek advice from regional health departments (Municipal Health Services), who, in turn, can be assisted by the PRU. HRIG and vaccine are only provided by the National Vaccine Institute in Bilthoven. PMID- 18389876 TI - [Increasing pressure on physicians to speak or remain silent; recent legal developments with regard to duty of professional confidentiality and right of non disclosure]. AB - In principle, the provision of data to third parties, without the patient's consent, is not allowed. There are recent legal developments with regard to the professional confidentiality and right of non-disclosure of the physician. Three subjects are explored in depth here, i.e. the developments with regard to professional confidentiality in child abuse and the deceased, and with regard to the Electronic Patient File. Both jurisprudence and two recent bills (Youth Care Act and Social Security Number in Health Care Bill) show that pressure on physicians and institutions to breach the duty of professional confidentiality is increasing. Physicians and their organisations should focus more attention on these problems by education, protocolling and discussion with The Netherlands Health Care Inspectorate and the Public Prosecution Service. PMID- 18389877 TI - [Safe illegal abortion would make an important contribution to decreasing maternal mortality and female suffering]. AB - Unsafe abortion is a major public health concern in many developing countries, contributing to a substantial proportion of maternal deaths. Increased legal access to abortion services is associated with improvement in mortality and morbidity. Safe illegal abortion by means of drugs is another possible way to prevent this unnecessary harm to women. The Dutch government, however, is now proposing legal changes that will diminish the access to medical abortion by general practitioners. PMID- 18389878 TI - [No reason to start treatment with statins in patients with moderate to severe heart failure]. AB - The 'Controlled rosuvastatin multinational trial in heart failure' (CORONA) recently reported that treatment with 10 mg of rosuvastatin per day has no significant effect on primary endpoints cardiovascular death, non-fatal myocardial infarction and non-fatal stroke in older patients with ischaemic systolic heart failure, despite a reduction in LDL-cholesterol of 45% to a level of 2.0 mmol/l. High sensitivity C-reactive protein was also reduced by 37%. The number of hospitalizations for cardiovascular disorders was significantly reduced. There were no more side effects in the rosuvastatin group than in the placebo group, even though the population consisted of elderly people with comorbidity and at risk for adverse drug interactions due to polypharmacy. There were no signs of any pleiotropic effects of statins in patients with heart failure. There seems to be no indication to start treatment with statins in patients with moderate to severe heart failure. PMID- 18389879 TI - [Pheochromocytomas and paragangliomas: implications of new insights for diagnosis and treatment]. AB - The recent discovery of pathogenic mutations in genes encoding for succinate dehydrogenase subunits has led to the realization that pheochromocytomas and paragangliomas are much more often hereditary than was previously thought. Due to periodic surveillance of patients at enhanced genetic risk and a general increase in the frequency of abdominal imaging, an ever increasing proportion of the pheochromocytomas and paragangliomas is now detected preclinically, without the classic symptoms and signs. The diagnosis ofa pheochromocytoma or paraganglioma can be confirmed by measurement of the plasma levels and 24-hour urinary excretion of catecholamines, in combination with imaging. The therapeutic strategy will depend on the localisation of the pheochromocytoma or paraganglioma, its solitary or multiple presence, the absence or presence of excessive catecholamine production, and the gene involved. PMID- 18389880 TI - [The effect of hormone therapy on cognitive function in patients with breast cancer]. AB - Preclinical and clinical studies suggest that oestrogens have an important role in brain functioning and cognitive ability. Given that hormone therapies for breast cancer reduce oestrogen levels or block oestrogen receptors, it is conceivable that these agents also influence cognitive function. Several small studies have been conducted to address this issue, but many of them are methodologically insufficient. The negative effects of oophorectomy and luteinising hormone-releasing hormone (LHRH) analogues on verbal memory and working memory have been demonstrated the most consistently, albeit only in small studies. Anastrozole and tamoxifen also appear to exert some negative effect on cognition, but well-designed studies are lacking. No data are available on the influence of the aromatase inhibitors exemestane and letrozole on cognitive function. Raloxifene, a drug that has no obvious advantages over tamoxifen and will likely not be developed further for breast cancer treatment, has no negative influence on cognitive functioning. It remains unclear whether the observed effects are transient or permanent, and to what extent age, menopausal status and duration of therapy influence the severity of cognitive effects. PMID- 18389881 TI - [New drugs; natalizumab]. AB - The drug natalizumab represents a new pharmacological approach in the treatment ofvery active relapsing-remitting multiple sclerosis (MS). It is a humanised murine monoclonal antibody and binds to an integrin on the surface oflymphocytes, thereby preventing them from transmigrating across the endothelium and causing inflammation in the nervous tissue. The drug has been shown to decrease the occurrence of relapses and progression of MS. A few severe adverse effects (such as the viral progressive multifocal leuko-encephalopathy) have been reported, and its clinical and long-term effects are not fully known at present. Therefore, further research is required to determine the role of natalizumab in clinical practice. PMID- 18389882 TI - [Diagnostic image (363). A basketball player with skin abnormalities]. AB - A 14-year-old male fanatic basketball player with a good skill of dunking presented with skin abnormalities on the right side of his back due to sports related striae, stretch marks. PMID- 18389883 TI - [The neurovascular costoclavicular compression syndrome exists]. AB - The neurovascular costoclavicular compression syndrome is rare. Physicians generally have little experience with this syndrome. It is difficult to provide evidence based on sound clinical science that proves the existence of the syndrome in relation to diagnosis and treatment. Nevertheless there are reasons to assume that this syndrome exists on the basis of clinical experience, particularly that of vascular surgeons. PMID- 18389884 TI - [The neurovascular costoclavicular compression syndrome does not exist]. AB - The term 'neurovascular compression syndrome' refers to a heterogeneous group of signs and symptoms thought to be generated by the compression of arteries, veins or nerves, which is caused by the anatomical relationships of muscles, ligaments and bony structures in the thoracic outlet. Most cases are related to nerve compression; arterial and venous compression accounts for 1-3% of cases. The term 'neurovascular compression syndrome', which does not refer to a single entity, is confusing and should be avoided. Rather, a clinical problem thought to arise in the thoracic outlet should be described in terms of the underlying arterial, venous or nerve impairment. Indications for surgical intervention are limited: there is no evidence to suggest that patients who undergo surgery fare better than those who receive non-surgical treatment. PMID- 18389885 TI - [Good results of early medicinal pregnancy termination in a general practice on Curacao]. AB - OBJECTIVE: To evaluate the results of early medicinal pregnancy termination. DESIGN: Prospective, non-blinded, clinical study. METHOD: Between 2001 and 2005, 371 women presented to a medium-sized solo general practice in the centre of Willemstad, Curacao, to request termination of an undesired pregnancy. Ofthese women, 74 first indicated a preference for abortion via curettage, and the remaining 297 chose to be treated medicinally. In the first trimester of the pregnancy, they were given methotrexate i.m. and misoprostol vaginally 3-5 days later, or only misoprostol vaginally when methotrexate was unavailable. The result was evaluated by abdominal echography two days after the treatment. RESULTS: The result of the medicinal pregnancy termination could be determined in 271 of the 297 women. In 219 (81%) of these, the pregnancy had been terminated after a single treatment, and in 29 (11%) success was achieved after a second vaginal administration of misoprostol. The total success rate of the medicinal pregnancy termination was 92% (248/271). Curettage was required in 23 women (8%) after the medicinal treatment had failed. There was no significant difference in success rate between treatment with the methotrexate-misoprostol combination and treatment with misoprostol alone. CONCLUSION: Medicinal pregnancy termination with methotrexate and misoprostol, or with misoprostol alone, was a safe method with a relatively small risk of an ongoing pregnancy. It was a good alternative for surgical abortion. PMID- 18389886 TI - [Serious intoxication with hydroxychloroquine, with haemodynamic instability: a case report supporting treatment with diazepam]. AB - A 37-year-old woman was admitted to the emergency room because of an autointoxication with hydroxychloroquine, leading to haemodynamic instability. Treatment consisted of the rapid administration of intravenous diazepam, after which the hypotension recovered rapidly even though no vasoactive medication was given. Treatment with diazepam has been advised in the Netherlands for many years in case of severe hydroxychloroquine intoxication, despite the fact that convincing evidence for its use is lacking. On the basis of the experience with the relevant cases, the administration of diazepam, 2 mg/kg initially followed by 2 mg/kg/24 hours as a continuous infusion, should certainly be considered for supportive treatment in the ICU in case of severe haemodynamic instability. PMID- 18389887 TI - [Churg-Strauss syndrome in a patient with asthma treated with montelukast]. AB - A 75-year-old woman with a history of asthma, rhinitis and nasal polyps was admitted due to petechial lesions on the lower left leg and weakness of the right foot. Six weeks prior to admission, she had started treatment with montelukast 10 mg daily. Based on the asthma, eosinophilia, mononeuritis of the right leg and a skin biopsy showing small vessel vasculitis with eosinophilic granulocytes, the patient was diagnosed with Churg-Strauss syndrome (CSS). After consulting with the pulmonologist, montelukast therapy was discontinued and replaced with a combined preparation of a parasympatholytic and a P2-sympathomimetic. The patient was also given prednisone 60 mg daily, which resulted in prompt clinical improvement and resolution of the eosinophilia. Development of CSS has been associated with the use of montelukast and should be considered in patients with asthma who develop new symptoms, such as neuritis, vasculitis of the skin or pulmonary infiltrates with an increase in eosinophilia during montelukast therapy. In these patients, treatment with montelukast should be discontinued, although whether a causal relationship exists between montelukast and CSS continues to be debated in the literature. PMID- 18389888 TI - [Orphanet: a European database for rare diseases]. AB - Orphanet is a European initiative that aims to improve the management and treatment of rare diseases. It comprises a database dedicated to information on rare diseases and orphan drugs, and offers services adapted to the needs of patients and their families, health professionals, and researchers. The database can be accessed through the website (www.orpha.net) and has some interesting options for searching, for example research projects, support groups or searching by clinical signs. Health professionals are encouraged to add activities concerning rare diseases to the database. PMID- 18389889 TI - [A memorable patient history]. PMID- 18389890 TI - [Diagnostic image (354). A man with neck complaints after a skiing accident]. PMID- 18389891 TI - [Effectiveness of endarterectomy for symptomatic stenosis of the internal carotid artery; more risk factors important than only the severity of the stenosis]. PMID- 18389892 TI - [Lidocaine: local anaesthetic with systemic toxicity]. PMID- 18389893 TI - [No HIV test: a chance missed]. PMID- 18389894 TI - Darunavir: pharmacokinetics and drug interactions. AB - Darunavir (TMC114) is a new HIV protease inhibitor that has demonstrated substantial antiretroviral activity against wild-type HIV-1 virus and multidrug resistant strains. Darunavir inhibits and is primarily metabolized by cytochrome P450 3A (CYP3A) isoenzymes and is coadministered with low-dose ritonavir (darunavir/r); ritonavir is an inhibitor of CYP3A isoenzymes and pharmacologically enhances darunavir, resulting in increased plasma concentrations and allowing for a lower daily dose. The t1/2 (terminal elimination half-life) of darunavir is 15 h in the presence of ritonavir. An extensive darunavir/r drug-drug interaction programme has been undertaken, covering a wide range of therapeutic areas. Studies conducted in HIV-negative healthy volunteers and in HIV-infected patients show that the potential for interactions is well characterized and the interactions are manageable. For most drugs investigated, no dose adjustments of darunavir/r or the co-administered drug are required. This article reviews all the pharmacokinetic and drug-drug interaction studies conducted to date for darunavir/r, providing guidance on how to co-administer darunavir/r with many other antiretroviral or non-antiretroviral medications commonly used in HIV-infected individuals. PMID- 18389895 TI - Health-related quality of life and tolerability in treatment-experienced HIV-1 infected patients on tipranavir versus comparator regimens. AB - BACKGROUND: Antiretroviral therapy including tipranavir boosted with ritonavir (TPV/r) has shown superior viral suppression and immunological response compared with comparator ritonavir-boosted protease inhibitor (CPI/r) regimens in treatment-experienced HIV-1-infected patients. This study assesses the influence of adverse events (AEs) on health-related quality of life (HRQOL) and change in HRQOL in patients treated with TPV/r versus CPI/r regimens. METHODS: Changes in HRQOL over 48 weeks were assessed using Medical Outcomes Study HIV Health Survey (MOS-HIV) data combined from two randomized, open-label, Phase III studies (RESIST-1 and RESIST-2). Generalized estimating equations (GEE) were used to compare physical health and mental health summary scores and 10 subscale scores, and to compare scores of patients with and without AEs. To compare AE incidences in the two treatment groups, AEs were exposure-adjusted. RESULTS: There were 984 patients in the HRQOL analysis. AE occurrence and severity resulted in significantly lower MOS-HIV scores across both treatment arms (P<0.05). Overall incidence of AEs was higher in the CPI/r versus TPV/r group (562.8 versus 514.4 per 100 patient-exposure years); treatment-related AEs were more frequent in the TPV/r group (75.0 versus 56.6 per 100 patient-exposure years). HRQOL was maintained in patients on TPV/r over 48 weeks of treatment across all summary and subscale scores. Compared with CPI/r, TPV/r was associated with a significant but small (SD<0.2) improvement in pain scores (+4.8 points; P<0.05). CONCLUSIONS: HRQOL was maintained across both summary and all subscale scores from baseline to 48 weeks in the TPV/r and CPI/r treatment arms, despite the incidence of treatment-related AEs. PMID- 18389896 TI - Contribution of mitochondrial dysfunction and oxidative stress to cellular premature senescence induced by antiretroviral thymidine analogues. AB - OBJECTIVES: Treatment of HIV-infected patients is associated with early onset of aging-related comorbidities. Some of the adverse effects of antiretroviral therapy have been attributed to the mitochondrial toxicity of nucleoside reverse transcriptase inhibitors (NRTI), and it is of note that mitochondrial dysfunction and oxidative stress are involved in the aging processes. In this regard, we examined whether NRTIs could accelerate the senescence of cultured cells. METHODS: Human fibroblasts were exposed to NRTIs from culture passage 1 to 14. Cytochrome c-oxidase (COX) subunits 2 and 4, mitochondrial potential and mass, and reactive oxygen species (ROS) were quantified at each passage. Proliferation, cell-cycle arrest, senescence-associated beta-galactosidase activity, and morphology were assessed in parallel. Mitochondrial and senescence markers were assessed in cultured murine preadipocytes and in fat samples from lipodystrophic HIV-infected patients. RESULTS: Stavudine and zidovudine induced mitochondrial dysfunction and increased ROS levels in fibroblasts at early culture passages, while cell division gradually slowed. At passages 8-12, fibroblasts exposed to stavudine or zidovudine but not abacavir, didanosine, lamivudine and tenofovir were senescent, on the basis of p16(INK4) and p21(WAF-1) protein expression, cell morphology and senescence-associated-beta-galactosidase activity. Senescence markers and COX2 underexpression were also found in 3T3-F442A preadipocytes exposed for 7 weeks to stavudine or zidovudine, but not lamivudine, and in adipose tissue samples from lipodystrophic HIV-infected patients on antiretroviral regimens containing stavudine or zidovudine. CONCLUSIONS: Mitochondrial changes and oxidative damage could partly explain the premature senescence of fibroblasts and adipose cells induced by stavudine and zidovudine. This suggests that thymidine analogues might be involved in the early aging related diseases observed in some HIV-infected patients taking antiretroviral drugs. PMID- 18389897 TI - The prophylactic administration of a monoclonal antibody against human metapneumovirus attenuates viral disease and airways hyperresponsiveness in mice. AB - BACKGROUND: Human metapneumovirus (hMPV) is one of the most frequent causes of respiratory tract infections in children. Our objective was to assess the prophylactic benefit of a monoclonal antibody (mAb) against the hMPV fusion protein in a murine model. METHODS: BALB/c mice received one intramuscular injection of either 5 or 10 mg/kg of mAb 338 (MedImmune, Inc.) and were infected intranasally 24 h later with 1x10(8) TCID50 (50% tissue culture infectious dose) of hMPV. On days 5 and 42 post-infection, lung samples were collected for determination of viral titres and for histopathological studies. Pulmonary function was characterized by plethysmography. RESULTS: Mean lung viral titres were significantly lower in mice treated with 5 or 10 mg/kg of mAb 338 compared with infected controls on day 5 (283, 45.6 and 1.49x10(5) TCID50/g, respectively; P<0.05). Similarly, lung viral RNA copies were significantly reduced in treated mice on day 42 (292, 101 and 607 copies per 0.01 g of lungs for mice that received 5 mg/kg, 10 mg/kg or no mAb, respectively; P<0.05). Histopathological changes characterized by important alveolar and interstitial inflammation were less severe in treated mice on days 5 and 42 compared with control. Airways obstruction was also significantly reduced in both treated groups on days 5 and 42, but development of hyperresponsiveness following the acute phase of infection was only significantly reduced in 10 mg/kg treated mice. CONCLUSIONS: Prophylactic administration of mAb 338 attenuates acute and late consequences of hMPV disease in this mouse model. PMID- 18389898 TI - Early therapy in HIV-1-infected children: effect on HIV-1 dynamics and HIV-1 specific immune response. AB - BACKGROUND: Perinatal HIV-1 infection is acquired in the milieu of a developing immune system, leading to high levels of uncontrolled viral replication. Few data have been reported that address the viral dynamics and immunological response in infants who initiated aggressive antiretroviral therapy (ART) shortly after birth. METHODS: Six HIV-1-infected infants who started ART within 3 months of age were studied. The median followup was 61 months. Plasma HIV-1 RNA, cell associated HIV-1 DNA, unspliced and multiply spliced HIV-1 mRNAs, HIV-1 antibodies, and CD4+ and CD8+ T-cell subsets were assessed in sequential peripheral blood samples. HIV-1 cellular immune response was measured by EliSpot assay. RESULTS: All children showed a decline in plasma viraemia to undetectable levels. HIV-1 DNA persisted in four children, but only two of these had detectable HIV-1 mRNA. All viral parameters remained persistently negative in two children. Only two children produced HIV-1 antibodies, while the others, after having lost maternal antibodies, remained seronegative. No HIV-1 cellular immune response was observed in any child. Therapy interruption was performed in two children: one HIV-1-seropositive and one HIV-1-seronegative with persistently undetectable levels of all viral parameters. Rebound of HIV-1 plasma viraemia in the seronegative child was more rapid and higher than that observed in the seropositive child. CONCLUSIONS: Early ART treatment in infants modifies the natural course of infection by controlling HIV-1 replication and reducing viral load to below the threshold levels required for onset of HIV-1 immune response, but does not prevent the establishment of a reservoir of latently infected cells that precludes virus eradication. PMID- 18389899 TI - Sequential combination therapy leads to biochemical and histological improvement despite low ongoing intrahepatic hepatitis B virus replication. AB - BACKGROUND: We previously reported that 48 weeks of combination therapy with pegylated interferon-alpha2b (PEG-IFN-alpha2b) and adefovir dipivoxil (ADV) in patients with chronic hepatitis B led to marked decreases of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) (-2.4 log10 copies/ml). Combination therapy was followed by 96 weeks of ADV monotherapy. METHODS: Here, we report on the final outcome after 144 weeks of sequential antiviral treatment. Twenty-four patients were analysed, triplet liver biopsies (taken at baseline, week 48 and week 144) were available from 16 patients. RESULTS: At week 144, 12/15 patients positive for hepatitis B virus e antigen (HBeAg) had lost HBeAg, alanine transaminase (ALT) levels were normal in 23 patients (96%), median serum HBV DNA had decreased by -4.9 log10 copies/ml and was undetectable (<100 copies/ml) in 11/24 individuals (46%). Median total intrahepatic HBV DNA had decreased by -2.2 log. Although no further significant cccDNA changes occurred between week 48 and week 144, two years of ADV monotherapy proved capable of controlling cccDNA levels in most patients. Analysis of intrahepatic HBV DNA species demonstrated that combination therapy with PEG-IFN-alpha2b and ADV inhibited viral productivity by 99% and subsequent ADV monotherapy by 76%, respectively. Virus suppression to undetectability within the first 12 weeks of treatment was strongly associated with long-term virological response and HBeAg and hepatitis B virus surface antigen HBsAg seroconversion. Histological improvement was determined in 11/16 patients at week 144. Two patients developed ADV resistance during the third year of treatment. CONCLUSIONS: Reduction of intrahepatic viral load achieved after 48 weeks of combination therapy with PEG-IFN-alpha2b and ADV was maintained in the following 96 weeks of ADV monotherapy and translated into long-term clinical benefit for most of the treated patients. PMID- 18389900 TI - Effect of pioglitazone on HIV-1-related lipodystrophy: a randomized double-blind placebo-controlled trial (ANRS 113). AB - BACKGROUND: Although thiazolidinediones have been shown to increase subcutaneous fat in congenital lipodystrophy, rosiglitazone did not show convincing results in HIV lipoatrophy. We assess a potential specific effect of pioglitazone in this setting. METHODS: One-hundred and thirty HIV-1-infected adults with self-reported lipoatrophy confirmed by physical examination were randomized to receive pioglitazone 30 mg once daily (n=64) or placebo (n=66) for 48 weeks. Changes in limb fat between weeks 0 and 48 were measured using dual-energy Xray absorptiometry. Subcutaneous and visceral fat was measured by single-slice computed tomography; fasting plasma measurements of glucose, insulin and lipids levels were recorded. RESULTS: Limb fat increased by 0.38 kg in the pioglitazone group and 0.05 kg in the placebo group at week 48 (mean difference 0.33 kg, 95% confidence interval [CI] 0.10-0.56; P=0.051) by intention-to-treat analysis. In patients not receiving stavudine, an increase of 0.45 kg versus 0.04 kg was observed (mean difference, 0.40 kg, 95% CI 0.12-0.69; P=0.013), but this was not seen in patients on stavudine (n=36; P=0.404). Overall, there was no significant difference in subcutaneous abdominal fat or in visceral fat areas on computed tomography at L4 vertebra. The lipid profile was not significantly different at week 48 except for levels of high-density lipoprotein cholesterol, which was improved in the pioglitazone group (+0.08 mmol/l versus -0.08; P=0.005). CONCLUSIONS: Pioglitazone 30 mg once daily for 48 weeks improved limb fat atrophy in antiretroviral-treated HIV-1-infected patients, although clinical benefits were not perceived by the patients. Treatment did lead to a favourable lipid profile, however, suggesting that this thiazolidinedione should be considered in the context of HIV-related lipoatrophy. PMID- 18389901 TI - Self-reported non-adherence to antiretroviral therapy repeatedly assessed by two questions predicts treatment failure in virologically suppressed patients. AB - BACKGROUND: The aim of this study was to explore the predictive value of longitudinal self-reported adherence data on viral rebound. METHODS: Individuals in the Swiss HIV Cohort Study on combined antiretroviral therapy (cART) with RNA <50 copies/ml over the previous 3 months and who were interviewed about adherence at least once prior to 1 March 2007 were eligible. Adherence was defined in terms of missed doses of cART (0, 1, 2 or >2) in the previous 28 days. Viral rebound was defined as RNA >500 copies/ml. Cox regression models with time-independent and -dependent covariates were used to evaluate time to viral rebound. RESULTS: A total of 2,664 individuals and 15,530 visits were included. Across all visits, missing doses were reported as follows: 1 dose 14.7%, 2 doses 5.1%, >2 doses 3.8% taking <95% of doses 4.5% and missing > or =2 consecutive doses 3.2%. In total, 308 (11.6%) patients experienced viral rebound. After controlling for confounding variables, self-reported non-adherence remained significantly associated with the rate of occurrence of viral rebound (compared with zero missed doses: 1 dose, hazard ratio [HR] 1.03, 95% confidence interval [CI] 0.72-1.48; 2 doses, HR 2.17, 95% CI 1.46-3.25; >2 doses, HR 3.66, 95% CI 2.50-5.34). Several variables significantly associated with an increased risk of viral rebound irrespective of adherence were identified: being on a protease inhibitor or triple nucleoside regimen (compared with a non-nucleoside reverse transcriptase inhibitor), >5 previous cART regimens, seeing a less-experienced physician, taking co medication, and a shorter time virally suppressed. CONCLUSIONS: A simple self report adherence questionnaire repeatedly administered provides a sensitive measure of non-adherence that predicts viral rebound. PMID- 18389902 TI - Prospective cohort study of HIV post-exposure prophylaxis for sexual assault survivors. AB - BACKGROUND: There is a lack of standardized programs for HIV counselling and post exposure prophylaxis (PEP) in the setting of sexual assault. METHODS: We conducted an 18-month prospective cohort study assessing universal HIV counselling for all sexual assault survivors presenting to 18 Ontario Sexual Assault Treatment Centres. HIV PEP was universally offered to those at risk of HIV infection (high risk or unknown risk) presenting < or =72 h after the assault, using Combivir one pill and Kaletra three capsules twice a day for 28 days. Those who accepted HIV PEP were monitored via a schedule of frequent follow ups. The primary outcomes were acceptance and completion rates, and their predictors were determined using multivariable logistic regression. Adverse events (AE) were categorized using a standardized toxicity grading system. RESULTS: Of the 900 evaluable participants eligible for PEP, 798 (69 at high risk and 729 at unknown risk) were offered treatment. Acceptance rates were 66.7% (n=46) and 41.3% (n=301) for participants at high risk and unknown risk, respectively. Participants at high risk were 2.2 times more likely to accept PEP than those at unknown risk (adjusted odds ratio 2.2; 95% confidence interval 1.2 4.0; P=0.01). Overall, 23.9% high-risk (n=11) and 33.2% unknown-risk participants (n=100) completed PEP (P=0.20). Predictors of acceptance and completion included assault by a stranger and participant anxiety. AEs were common, with 77.1% of participants reporting grade 2-4 symptoms. CONCLUSION: A province-wide standardized program of universal HIV counselling and offering of PEP to sexual assault survivors with frequent follow up was successfully implemented and feasible. PMID- 18389904 TI - Antiretroviral-associated portal hypertension: a new clinical condition? Prevalence, predictors and outcome. AB - BACKGROUND: Cases of severe unexplained liver disease in HIV-infected individuals have recently been reported and are often associated with exposure to didanosine (ddl) and nodular regenerative hyperplasia. Herein, we examine the clinical outcome following ddl removal. METHODS: From 3,300 HIV-infected patients attending three clinics since 2004, all who exhibited persistently elevated aminotransferases and/or significant liver fibrosis in the absence of any known cause of liver damage were identified. RESULTS: Thirty-two individuals (prevalence approximately 1%) met the inclusion criteria - all were on antiretroviral therapy. Of these, 84% were male and 68% had acquired HIV through homosexual contact. Liver biopsy was performed in 12, of whom three showed nonspecific advanced liver fibrosis, two nodular regenerative hyperplasia and three showed only periportal fibrosis. On follow up, nine patients developed episodes of hepatic decompensation, mainly as a consequence of portal hypertension; in eight cases (25%) portal thrombosis was diagnosed. No association was found with plasma HIV RNA or CD4+ T-cell count. All patients but three had been exposed to ddl for a median of 44 months; removal of ddl in 27 was followed 12 months later by improvement in clinical and laboratory parameters in 13 (48%) patients. Finally, a trend towards liver fibrosis improvement was recognised using FibroScan. CONCLUSIONS: Idiopathic persistent liver enzyme elevations in HIV-infected individuals are often associated with cirrhotic and non-cirrhotic portal hypertension. Although this is a relatively rare condition, prolonged exposure to ddl seems to play a pathogenic role and removal of the drug is associated with clinical and laboratory improvements. PMID- 18389903 TI - Does treatment of hepatitis B virus (HBV) infection reduce hepatitis delta virus (HDV) replication in HIV-HBV-HDV-coinfected patients? AB - BACKGROUND: Hepatitis delta virus (HDV) has a unique replication process that requires coinfection with hepatitis B virus (HBV). Treatment is currently limited to interferon therapy. The role of potent nucleos(t)ide analogues active against HBV has not been well examined in chronic delta hepatitis (CDH). METHODS: HIV positive patients with CDH attending our hospital were identified and longitudinally studied. Serum HBV DNA, HDV RNA and HIV RNA, treatment regimens, and biochemical and serological markers were assessed at yearly intervals. Liver fibrosis was measured by transient elastography during the last 2 years. RESULTS: Sixteen patients were identified and treated with anti-HBV therapy (median time 6.1 years). The majority were male and previous intravenous drug users. Median baselines were: HDV RNA 7 log10 copies/ml, HIV RNA 1.7 log10 copies/ml, HBV DNA 1.1 log10 IU/ml and alanine aminotransferase (ALT) 98 IU/ml. A significant correlation was found between HDV RNA and HBV DNA (r=0.226, P=0.015), aspartate aminotransferase (r=0.430, P<0.0001), ALT (r=0.441, P<0.0001) and hepatitis B surface antigen (HBsAg) (r=0.557, P<0.0001). Overall, 13 patients showed a reduction in HDV viraemia and ALT levels, and three of them achieved undetectable HDV RNA and normal ALT levels. CONCLUSION: Patients undergoing successful anti HBV therapy with potent nucleos(t)ide analogues seem to indirectly benefit from suppression of HDV replication, albeit not very efficiently. Hypothetically, a significant and sustained reduction in serum HDV RNA might only be seen when a reduction in HBV covalently closed circular DNA or HBV surface antigen is achieved, which may require long periods of successful anti-HBV therapy. To our knowledge, this is the first evidence of the benefit of potent anti-HBV nucleos(t)ide analogue therapy in CDH. PMID- 18389905 TI - In vitro and in vivo protection against the highly pathogenic H5N1 influenza virus by an antisense phosphorothioate oligonucleotide. AB - BACKGROUND: Current vaccination strategies and antiviral drugs only provide limited protection against influenza virus infection. In this study, we investigated the use of a novel antisense oligonucleotide (named IV-AS), which is specific for the 5'-terminal conserved sequence found in all eight viral RNA segments of influenza A virus. METHODS: The activity of IV-AS was monitored both in vitro, in Madin-Darby canine kidney (MDCK) cells, and in vivo using a mouse model. IV-AS was given intranasally to H5N1-infected mice once daily for 6 days starting 6 h after infection. A three-base mismatch of IV-AS was used as a control. RESULTS: IV-AS inhibited influenza virus A induced cytopathic effects in MDCK cells with the 50% effective concentration (EC50) ranging from 2.2 to 4.4 microM. IV-AS was effective against H5N1 virus in preventing death, lessening weight reduction, inhibiting lung consolidation and reducing lung virus titres. Dosages of 40 and 60 mg/kg/day provided 40% and 60% survival rates and prolonged mean survival days in comparison with the infected control group (P<0.05). The lung index in mice treated with IV-AS, at a dose of 20, 40 or 60 mg/kg/day, had been inhibited on day 4 or 6 (P<0.05 or P<0.01); virus titres in lung had declined to 2.42, 1.51 and 1.54 log10 TCID50/g of lung, respectively, whereas the yields in the infected control mice were 6.00 log10 TCID50/g of lung. CONCLUSIONS: Our results suggest that the 5'-terminal conserved region of influenza A virus RNA segments can be targeted using antisense technology; therefore, IV-AS is a potential drug for prophylaxis and control of influenza virus infections. PMID- 18389906 TI - Gln151 of HIV-1 reverse transcriptase acts as a steric gate towards clinically relevant acyclic phosphonate nucleotide analogues. AB - BACKGROUND: In the treatment of HIV, the loose active site of the HIV-1 reverse transcriptase (RT) allows numerous nucleotide analogues to act as proviral DNA 'chain-terminators'. Acyclic nucleotide phosphonate analogues (ANPs) represent a particular class of nucleotide analogue that does not possess a ribose moiety. The structural basis for their substrate efficiency regarding viral DNA polymerases is poorly understood. METHODS: Pre-steady-state kinetics on HIV-1 RT together with molecular modelling, were used to evaluate the relative characteristics of both the initial binding and incorporation into DNA of three different ANP diphosphates with progressively increasing steric demands on the acyclic linker: adefovir-diphosphate (DP), tenofovir-DP, and cidofovir-DP. RESULTS: The increase of steric demand in ANPs induced a proportional loss of the binding affinity to wild-type HIV-1 RT (Kd cidofovir-DP>>Kd tenofovir-DP>Kd adefovir-DP approximately Kd dNTPs), consistent with the lack of HIV-1 inhibitory activity for cidofovir. We show that, starting from adefovir-DP, the steric constraints mainly map to Gln151, as its mutation to alanine provides cidofovir DP sensitivity. Interactions between the Gln151 residue and the methyl group of tenofovir-DP further increase with the mutation Gln151Met, resulting in a specific discrimination and low-level resistance to tenofovir-DP. This alteration is the result of a dual decrease in the binding affinity (Kd) and the catalytic rate (k(pol)) of incorporation of tenofovir-DP. By contrast, the tenofovir resistance mutation K65R induces a broad 'k(pol)-dependent' nonspecific discrimination towards the three ANPs. CONCLUSIONS: Overall, our results show that the efficiency of ANPs to compete against natural nucleotides as substrates for RT is determined by their close interaction with specific amino acids such as Gln151 within the RT active site. These results should help us to map and predict ANP sensitivity determinants in cellular and viral DNA polymerase active sites for which the understanding of different ANP sensitivity patterns are of medical importance. PMID- 18389907 TI - Passive immunization protects cynomolgus macaques against Puumala hantavirus challenge. AB - BACKGROUND: Hantaviruses cause two severe and often fatal human diseases: haemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). Presently, there is no effective prevention available for HFRS or HPS. Here, we studied the effect of passive immunization on the course of infection in cynomolgus macaques challenged with wild-type Puumala hantavirus (PUUV-wt). METHODS: A pool of serum drawn from previously PUUV-wt-infected monkeys was used for immunization; a pool of serum from the same monkeys that was obtained before infection was used as a control. Immunizations were administered 3 days before and 15 days after challenge with PUUV-wt. After challenge, monkeys were sampled once a week and analysed for PUUV-infection markers. RESULTS: All three monkeys treated with non-immune serum became positive for PUUV RNA in plasma and showed PUUV nucleocapsid-specific immunoglobin M (IgM) responses after challenge. In contrast, no PUUV RNA or anti-PUUV-specific IgM response was detected in the three passively immunized monkeys. As seen in PUUV-infected humans, the control monkeys showed a marked decrease in the amount of platelets and increased levels of creatinine, interleukin (1L)-6, IL-10, and tumour necrosis factor (TNF) after inoculation. In contrast, no marked changes in the amount of platelets were observed in the immunized monkeys and they did not show increased levels of creatinine, IL-6, IL-10 and TNF after virus challenge. CONCLUSION: The results show that passive immunization in monkeys, using serum from previously hantavirus infected monkeys, can induce sterile protection and protect against pathogenesis. Convalescent-phase antibodies may represent a potential therapy that can induce immediate protection against HFRS and HPS. PMID- 18389908 TI - Low rate of emergence of nevirapine and lamivudine resistance after post-partum interruption of a triple-drug regimen. AB - INTRODUCTION: Emergence of nevirapine (NVP) resistance may be a consequence of its use in monotherapy to prevent HIV mother-to-child transmission (MTCT). The aim of this study was to evaluate the emergence of strains resistant to NVP and lamivudine (3TC) after discontinuation of antiretroviral therapy (ART) with 3TC/zidovudine (ZDV)/NVP. METHODS: Twenty pregnant women (ART-naive or preexposed only to ZDV), to whom 3TC/ZDV/NVP was prescribed as MTCT prophylaxis, were studied. They received ART for a median of 4 months with median viral load (VL) at labour <50 copies/ml. Samples were collected between 1 and 15 months (median: 3 months) after ART interruption. Sequence-selective real-time PCR (SPCR), which quantifies minority viral populations containing K103N, Y181C and M184V mutations, and standard genotypic sequencing were assayed. RESULTS: No mutations associated with resistance to 3TC or NVP were found by standard population sequencing. Analysis of K103N by SPCR showed that 35% of the patients contained < or =0.1% of viruses carrying either the AAC or AAT mutations. For Y181C mutation, 10% of the patients contained <0.5% of viruses with TGT codon change. For M184V mutation, one patient contained 6.2% of virus with GTG mutation and 13 patients (65%) contained <0.9% of mutated viruses. Four women were re-exposed to 3TC/ZDV/NVP and achieved HIV VL <50 copies/ml. No perinatal transmission occurred in any of the 22 births. CONCLUSIONS: NVP associated with ZDV/3TC as a regimen to prevent MTCT may involve a low risk for the selection of antiretroviral-resistant strains and may not jeopardize the use of these same drugs for future treatment. PMID- 18389909 TI - Characterization of drug resistance in antiretroviral-treated patients infected with HIV-1 CRF02_AG and AGK subtypes in Mali and Burkina Faso. AB - BACKGROUND: In a multicentred cohort of patients on antiretroviral therapy (ART) in Burkina Faso and Mali, we analysed the prevalence of HIV drug resistance mutations in patients failing a modified directly observed therapy (mDOT) protocol. METHODS: Patients on ART >6 months and with viral load (VL) >500 copies/ml were enrolled in a mDOT protocol. Genotypic resistance testing was performed on pre- and post-mDOT plasma samples of patients who still had VL >500 copies/ml after mDOT. RESULTS: Eight hundred and one patients from seven sites participated in the study. One hundred and thirteen patients (14.1%) had VL >500 copies/ml. Most patients were treated with lamivudine along with zidovudine or stavudine and efavirenz or nevirapine. Genotypes were available for 46 patients. The predominant HIV-1 subtypes were CRFO2_AG in 26 (56.5%) and AGK/K/AK in 12 (26.1%) patients. The prevalence of drug resistance mutations by class were as follows for nucleoside reverse transcriptase inhibitors: 1841/V (82.6%), 215Y/F (32.6%), 219E/Q (19.6%), 70R (19.6%), 67N (21.7%), 41L (15.2%) and 151M(2.2%). For non-nucleoside reverse transcriptase inhibitors the prevalence was: 103N (50%) and 181C/I (19.6%). Phylogenetic analysis showed that, although the genetic distances were small among isolates, there was no clustering of a particular subtype in a specific region and that the high prevalence of AGK subtype in our drug-resistant population was not due to a circulating resistant strain. CONCLUSION: Although CRFO2_AG is the dominant clade in the Burkina Faso/Mali region, isolates with subtype K reverse transcriptase were frequent in our cohort. Drug resistance mutation pathways in subtype K reverse transcriptase need to be further evaluated in a larger cohort of non-B HIV-infected individuals. PMID- 18389910 TI - [Article 17 of the Medical Law and the nurse anesthetist: logical argument for increasing the number of anesthetists]. PMID- 18389912 TI - Toll-free number for reporting adverse events on labeling for human drug products. Interim final rule. AB - The Food and Drug Administration (FDA) is issuing an interim final rule to codify the provisions of the proposed rule entitled "Toll-Free Number for Reporting Adverse Events on Labeling for Human Drug Products" (69 FR 21778, April 22, 2004) (the toll-free number proposed rule or proposed rule) that, under the Food and Drug Administration Amendments Act of 2007 (FDAAA), became effective by operation of law on January 1, 2008. This interim final rule requires the addition of a statement on the labeling of certain human drug products for which an application is approved under the Federal Food, Drug, and Cosmetic Act (the act). The added statement includes a toll-free number and advises that the number is to be used only for reporting side effects and is not intended for medical advice (the side effects statement). As mandated by FDAAA, this interim final rule does not apply to over-the-counter drug products approved as new drugs under the act if the product packaging includes a manufacturer's or distributor's toll-free number for reporting complaints. PMID- 18389913 TI - Both praying and playing: "Muscular Christianity" and the YMCA in north-east county Durham. PMID- 18389915 TI - Medicare program; revisions to payment policies under the Physician Fee Schedule, and other part B payment policies for CY 2008; delay of the date of applicability of the revised anti-markup provisions for certain services furnished in certain locations (Sec. 414.50). Final rule. AB - This final rule delays until January 1, 2009 the applicability of the anti-markup provisions in Sec. 414.50, as revised at 72 FR 66222, except with respect to the technical component of a purchased diagnostic test and with respect to any anatomic pathology diagnostic testing services furnished in space that: Is utilized by a physician group practice as a "centralized building" (as defined at Sec. 411.351 of this chapter) for purposes of complying with the physician self referral rules; and does not qualify as a "same building" under Sec. 411.355(b)(2)(i) of this chapter. PMID- 18389914 TI - "Journeys into poverty kingdom": complete participation and the British vagrant, 1866-1914. PMID- 18389916 TI - Authorized sources of narcotic raw materials. Final rule. AB - The Drug Enforcement Administration (DEA) is amending the list of non-traditional countries authorized to export narcotic raw materials (NRM) to the United States by removing Yugoslavia and adding Spain. This rule provides DEA registered importers with another potential source from which to purchase NRM that are used in the production of controlled substances for medical purposes in the United States. PMID- 18389917 TI - Revisions to the requirements applicable to blood, blood components and source plasma; confirmation of effective date and technical amendment. Direct final rule; confirmation of effective date and technical amendment. AB - The Food and Drug Administration (FDA) is confirming the effective date of February 19, 2008, for the direct final rule that appeared in the Federal Register of August 16, 2007 (72 FR 45883). The direct final rule amends the biologics regulations by removing, revising, or updating specific regulations applicable to blood, blood components and Source Plasma to be more consistent with current practices in the blood industry and to remove unnecessary or outdated requirements. In addition, FDA is making technical amendments to the biologics regulations in response to comments received on the direct final rule. PMID- 18389918 TI - Medicare program; prior determination for certain items and services. Final rule. AB - This final rule establishes a process for Medicare contractors to provide eligible participating physicians and beneficiaries with a determination of coverage relating to medical necessity for certain physicians' services before the services are furnished. This rule is intended to afford the physician and beneficiary the opportunity to know the financial liability for a service before expenses are incurred. This final rule establishes reasonable limits on physicians' services for which a prior determination of coverage may be requested and discusses generally our plans for establishing the procedures by which those determinations may be obtained. This rule also responds to public comments on the August 30, 2005 proposed rule. PMID- 18389919 TI - Medicare program; Medicare secondary payer (MSP) amendments. final rule. AB - On February 24, 2006, we published an interim final rule with comment period in the Federal Register that implemented amendments to the Medicare Secondary Payer (MSP) provisions under Title III of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA). The MMA clarified the MSP provisions regarding the obligations of primary plans and primary payers, the nature of the insurance arrangements subject to the MSP rules, the circumstances under which Medicare may make conditional payments, and the obligations of primary payers to reimburse Medicare. In this final rule, we are finalizing several clarifications made to the MSP provisions. In addition, we are responding to public comments on the February 24, 2006 interim final rule with comment period that pertain to these MSP provisions. PMID- 18389920 TI - Medicaid program; health care-related taxes. AB - This final rule revises the collection threshold under the regulatory indirect guarantee hold harmless arrangement test to reflect the provisions of the Tax Relief and Health Care Act of 2006. When determining whether there is an indirect guarantee under the 2-prong test for portions of fiscal years beginning on or after January 1, 2008 and before October 1, 2011, the allowable amount that can be collected from a health care-related tax is reduced from 6 to 5.5 percent of net patient revenues received by the taxpayers. This final rule also clarifies the standard for determining the existence of a hold harmless arrangement under the positive correlation test, Medicaid payment test, and the guarantee test (with conforming changes to parallel provisions concerning hold harmless arrangements with respect to provider-related donations); codifies changes to permissible class of health care items or services related to managed care organizations as enacted by the Deficit Reduction Act of 2005; and, removes obsolete transition period regulatory language. PMID- 18389921 TI - Food labeling: health claims; soluble fiber from certain foods and risk of coronary heart disease. Interim final rule. AB - The Food and Drug Administration (FDA) is amending the health claim regulation entitled "Soluble fiber from certain foods and risk of coronary heart disease (CHD)" to add barley betafiber as an additional eligible source of beta-glucan soluble fiber. Barley betafiber is the ethanol precipitated soluble fraction of cellulase and alpha-amylase hydrolyzed whole grain barley flour. FDA is taking this action in response to a health claim petition submitted by Cargill, Inc. FDA previously concluded that there was significant scientific agreement that a claim characterizing the relationship between beta-glucan soluble fiber of certain whole oat and whole grain barley products and CHD risk is supported by the totality of publicly available scientific evidence. Based on the totality of publicly available scientific evidence, FDA now has concluded that in addition to certain whole oat and whole grain barley products, barley betafiber is also an appropriate source of beta-glucan soluble fiber. Therefore, FDA is amending the health claim regulation entitled "Soluble fiber from certain foods and risk of CHD" to include barley betafiber as another eligible source of beta-glucan soluble fiber. PMID- 18389922 TI - Considerations for inpatient versus outpatient treatment with antipsychotics. PMID- 18389923 TI - Can oral cancer occur in someone without known risk? PMID- 18389924 TI - Is the message clear? Talking with your patient about oral cancer screening. PMID- 18389925 TI - Drug-drug interactions with antipsychotics. PMID- 18389926 TI - What do I say to my patient who needs a biopsy? PMID- 18389927 TI - Acute and long-term adverse effects of antipsychotics. PMID- 18389928 TI - Impact of medical comorbidity on medication management of schizophrenia. PMID- 18389929 TI - Augmentation strategies in the treatment of major depressive disorder. Clinical considerations with atypical antipsychotic augmentation. PMID- 18389930 TI - ADHD in adults--a familiar disease with unfamiliar challenges. PMID- 18389931 TI - Case of the month. Bisphosphonate related osteochemonecrosis of the jaws. PMID- 18389932 TI - Oral cancer screening and prevention. PMID- 18389933 TI - This little one smiled, all the way home. PMID- 18389934 TI - Special needs get special attention. PMID- 18389935 TI - Imaging as a Biomarker: Standards for Change Measurements in Therapy workshop summary. AB - Biomarkers are biological indicators of disease or therapeutic effects that can be measured by in vivo biomedical/molecular imaging, as well as other in vitro or laboratory methods. Recent work has shown that biomedical imaging can provide an early indication of drug response by use of x-ray, computed tomography (CT), positron-emission tomography/CT (PET/CT), or magnetic resonance imaging (MRI). There are three primary sources of uncertainty in using imaging as a biomarker: 1) the biological variability, 2) the variability associated with the clinicians interpreting the images, and 3) the physical measurement variability associated with image data collection and analysis across the same or different imaging platforms. Although biological variability is a large source of error, the physical uncertainty often significantly reduces the robustness of the imaging methods and the clinical decision tools required for quantitative measurement of therapy response over time. Physical and biological measurement uncertainties may be addressed prior to designing a clinical trial and thus help in reducing the case size and cost of a clinical trial associated with a drug submission to the U.S. Food and Drug Administration (FDA). The National Institute of Standards and Technology (NIST) has been approached over the last few years by several industry and medical stakeholders to address the physical sources of measurement uncertainty. NIST's initial research discovered that the characterization of measurement uncertainty poses many complex metrology and standardization problems on a scale that appears to need significant collaboration across the different medical imaging stakeholders. Many of the issues are similar to other scientific domains that NIST has addressed as part of its mission to provide metrology standards to enhance the competitiveness of U.S. industries. To better assess the measurement and standards needs for using imaging as a biomarker, NIST engaged leading representatives from many of the different imaging societies, the imaging, pharmaceutical and e-health and other health care stakeholders, as well as other key federal agencies (the National Institutes of Health Institutes and Centers [NIH ICs], and FDA) to organize and conduct a United States Measurement System (USMS) workshop: http://usms.nist.gov/workshops. The workshop entitled Imaging as a Biomarker: Standards for Change Measurements in Therapy, was thus held on September 14-15, 2006, at NIST in Gaithersburg, Maryland. (Workshop agenda, presentations. and final workshop report will be available at http://usms.nist.gov/workshops/bioimaging.htm.) PMID- 18389936 TI - Salmonella: methods and protocols. PMID- 18389937 TI - Ammonia assessment from agriculture: U.S. status and needs. AB - Recent studies suggest that human activities accelerate the production of reactive nitrogen on a global scale. Increased nitrogen emissions may lead to environmental impacts including photochemical air pollution, reduced visibility, changes in biodiversity, and stratospheric ozone depletion. In the last 50 yr, emissions of ammonia (NH3), which is the most abundant form of reduced reactive nitrogen in the atmosphere, have significantly increased as a result of intensive agricultural management and greater livestock production in many developed countries. These agricultural production practices are increasingly subject to governmental regulations intended to protect air resources. It is therefore important that an accurate and robust agricultural emission factors database exist to provide valid scientific support of these regulations. This paper highlights some of the recent work that was presented at the 2006 Workshop on Agricultural Air Quality in Washington, D.C. regarding NH3 emissions estimates and emission factors from agricultural sources in the U.S. and Europe. In addition, several best management practices are explored as the scientific community attempts to maximize the beneficial use of reactive nitrogen while simultaneously minimizing negative environmental impacts. PMID- 18389938 TI - Nitrous oxide emissions from a Northern Great Plains soil as influenced by nitrogen management and cropping systems. AB - Field measurements of N2O emissions from soils are limited for cropping systems in the semiarid northern Great Plains (NGP). The objectives were to develop N2O emission-time profiles for cropping systems in the semiarid NGP, define important periods of loss, determine the impact of best management practices on N2O losses, and estimate direct N fertilizer-induced emissions (FIE). No-till (NT) wheat (Triticum Aestivum L.)-fallow, wheat-wheat, and wheat-pea (Pisum sativum), and conventional till (CT) wheat-fallow, all with three N regimes (200 and 100 kg N ha(-1) available N, unfertilized control); plus a perennial grass-alfalfa (Medicago sativa L.) system were sampled over 2 yr using vented chambers. Cumulative 2-yr N2O emissions were modest in contrast to reports from more humid regions. Greatest N2O flux activity occurred following urea-N fertilization (10 wk) and during freeze-thaw cycles. Together these periods comprised up to 84% of the 2-yr total. Nitrification was probably the dominant process responsible for N2O emissions during the post-N fertilization period, while denitrification was more important during freeze-thaw cycles. Cumulative 2-yr N2O-N losses from fertilized regimes were greater for wheat-wheat (1.31 kg N ha(-1)) than wheat fallow (CT and NT) (0.48 kg N ha(-1)), and wheat-pea (0.71 kg N ha(-1)) due to an additional N fertilization event. Cumulative losses from unfertilized cropping systems were not different from perennial grass-alfalfa (0.28 kg N ha(-1)). Tillage did not affect N2O losses for the wheat-fallow systems. Mean FIE level was equivalent to 0.26% of applied N, and considerably below the Intergovernmental Panel on Climate Change mean default value (1.25%). PMID- 18389941 TI - [Insatiable challenge for eradication of leprosy]. PMID- 18389942 TI - Listen to your heart. PMID- 18389943 TI - The HRT dilemma. PMID- 18389944 TI - Solving insomnia. Chronic sleep problems are worth treating. PMID- 18389945 TI - Gain against the pain. Fibromyalgia knowledge and treatment have improved. PMID- 18389946 TI - What to do about HPV? PMID- 18389947 TI - The heart after breast cancer. PMID- 18389948 TI - A nasty bug breaks out. PMID- 18389949 TI - For frogs, a digital detour. PMID- 18389950 TI - [Neuro-muscular apparatus state in lumbosacral radiculopathy in miners]. AB - Based on clinical and electrophysiologic studies, the authors analysed neuro muscular apparatus of "spinal center--periphery" axis for miners with radicular pain caused by occupational lumbosacral radiculopathy. Findings are that constantly irritated receptors in lumbar motor segment during occupational activities alter habitual motor stereotype and cause specific compensatory muscular reactions, rearrangement of motor activity in segmental apparatus. PMID- 18389951 TI - [Physiologic shiftwise evaluation of respiratory system changes in miners engaged into underground coal mining]. AB - Functional shiftwise evaluation of physiologic respiratory parameters was conducted in 166 miners. The conclusion was that each separate occupation has specific requirements. PMID- 18389952 TI - [Hygienic evaluation of work activities in employees engaged into open-cast mining and chromium-ore concentration]. AB - Despite contemporary equipment usage on mining enterprises, work conditions are influenced by some hazards. PMID- 18389953 TI - [Cellular and cytogenetic evaluation of manganese-containing dust in experiment]. AB - Experimental studies helped to define biochemical, cytogenetic, cytochemical, genetic changes under influence of manganese-containing dust with concentrations characteristic for workplaces of open-cast mining. Cellular effect was accumulation of destructed cells in lungs, liver, adrenals. Cytogenetic effect also included increase in chromosomal aberattions and aneuploidy frequency in bone marrow. PMID- 18389954 TI - [Hygienic evaluation of work conditions in shift open-cast mining work in North Kazakhstan]. AB - Shift open-cast development and mining take place in Shubarkul coal deposit situated in semidesert with sharply continental climate. Health state and work capacity of the miners are influenced by climate, occupational and social environmental hazards. PMID- 18389955 TI - [On cognitive functions dynamics during operator activities]. AB - The authors analysed influence of sensory emotional exertion on higher psychic functions during operator activities, in accordance with age and length of service, occupationally specific features. PMID- 18389956 TI - [Clinical and functional characteristics of broncho-pulmonary system under exposure to chrysotile-asbestos dust]. AB - Early changes under exposure to chrysotile-asbestos dust include light arterial hypoxemia, lower bronchial permeability of small bronchi. Early asbestosis and dust bronchitis manifest with impaired pulmonary ventilation of mixed type, that is associated with moderate arterial hypoxemia in dust bronchitis. PMID- 18389957 TI - [Endothelial dysfunction development in miners]. AB - The authors studied vascular endothelium state in coal miners. Findings are lower endothelium-dependent and endothelium-independent vasodilatation, dysbalance of humoral markers--that prove endothelial dysfunction development and remodelling of vascular wall. PMID- 18389958 TI - [Automated database in analysis of pneumoconiosis prevalence in coal miners]. AB - Using simulator of forecasting dust occupational diseases with automated information system, the authors analysed risk of pneumoconioses development in coal miners. PMID- 18389959 TI - [International scientific-practical conference "Contemporary issues in occupational health and occupational diseases", Karaganda, September 20-21, 2007]. PMID- 18389960 TI - Health reform in Papua New Guinea and the Pacific. PMID- 18389961 TI - Prioritization in the Papua New Guinea health sector: progress towards a health medium-term expenditure framework. PMID- 18389962 TI - Implementing health policy and hospital standards. PMID- 18389963 TI - Health sector reform and the Health Management Reform Project in Fiji. PMID- 18389964 TI - Health sector reform in the Pacific: a Fijian divisional perspective. PMID- 18389965 TI - Health financing: the Fijian experience. PMID- 18389966 TI - The experience of health sector reform in Tonga. PMID- 18389968 TI - The experience of reform in the Samoa Ministry of Health. PMID- 18389967 TI - The experience of health reform in Vanuatu. PMID- 18389969 TI - The epidemiology of malaria in the Papua New Guinea highlands: 4. Enga Province. AB - Of all Papua New Guinea provinces, Enga has the largest proportion of people living at altitudes that preclude malaria transmission. However, the first systematic surveys in 1979 showed that malaria was endemic in lower-lying valleys to the north and east of the province. A series of new surveys conducted in both wet and dry seasons showed that these areas remain the main malaria focus in Enga. However, over the last 25 years the risk of malarial infections has increased substantially in areas < 1200 m (from 10% to 37-41%). In these low lying areas people acquire substantial antimalarial immunity and most infections are asymptomatic. However, people in villages in these areas had significantly lower mean haemoglobin levels (13.0 vs 14.0 g/dl, p < 0.001) than in areas above 1200 m, where overall prevalence rates (0-9%) have not changed much. In areas between 1200 and 1600 m epidemics with parasite prevalence rates in excess of 20% have been found to occur. Malaria was a significant cause of febrile illness only in endemic areas or during outbreaks. Although rarely used, sleeping under a bednet was associated with a significant reduction in risk of malaria infection (adjusted OR = 0.45, p = 0.01). On the other hand, sleeping in garden houses away from the main villages increased the risk of malaria infection (adjusted OR = 1.6, p = 0.03). Malaria control in outlying, malarious areas of Enga province could therefore be based on the distribution of long-lasting impregnated bednets, while at the same time addressing the additional risks posed by the high mobility of many of these populations through targeted health education. PMID- 18389970 TI - What do final year medical students at the University of Papua New Guinea think of psychiatry? AB - This study was conducted in 2004 to determine whether there was any difference between final year medical students who had rotated in psychiatry and those who had not, in terms of their preference for psychiatry as a career and their attitudes towards mental illness. A self-rated questionnaire was given to all the final year medical students at the University of Papua New Guinea. The results showed that the medical students in general had a negative attitude towards psychiatry as a career option and, although they were accepting of the mentally ill in a professional setting as colleagues or patients, they had a negative attitude towards close social contact with them as neighbours or as in-laws. Several students believed mental illness could be caused by sorcery or by spending much time with the mentally ill. Most students believed mental illness could be treated by prayer, one in five believed in the effectiveness of traditional healers and one in five did not believe modern medicine could treat mental illness. Apart from a reduction in stigma and in prejudice against a mentally ill neighbour, there was no significant difference in attitude between students who had rotated in psychiatry and those who had not. There was no significant difference in attitude between male and female respondents. There were, however, significant differences in attitude between students who had a positive family history of mental illness and those who did not. It was concluded that psychiatry was an unpopular choice for specialization and that students' attitudes towards mental illness were influenced more by their cultural beliefs and their family history of mental illness than by their rotation in psychiatry. These external variables that are independent of their medical training need to be considered during undergraduate medical training in order to optimize the provision of health care to the mentally ill in Papua New Guinea. PMID- 18389971 TI - Community screening for high blood pressure among adults in urban and rural Papua New Guinea. AB - This is a cross-sectional study conducted intermittently in Port Moresby, the National Capital District of Papua New Guinea, from 1996 to 1997; Mt Obree in Central Province in October 2000; Upper Strickland River in April 2001; and the Balopa Islands in Manus Province in December 2002. The aim of the study was to determine the prevalence of high blood pressure and identify possible risk factors for hypertension in the 'healthy' population in Port Moresby and the three rural communities. There were 1491 subjects surveyed, 704 males and 787 females. Their ages ranged from 20 to 84 years. Just over 6% of subjects were aged 65 years and above. There were 205 (14%) smokers and 340 (23%) betelnut chewers. The Central (rural) subjects were generally younger with the lowest mean systolic and diastolic blood pressures and lowest body mass index (BMI) in both males and females (no overweight or obesity). In Central and Strickland the mean systolic (SBP) and diastolic (DBP) blood pressures were lower and remained the same in all age groups, then in females decreased with age after 55 years. The Manus (rural) subjects were older with higher mean systolic and diastolic blood pressures and higher mean BMI, surprisingly similar to the urban population of Port Moresby. The mean systolic blood pressures in Port Moresby and Manus increased with age in both sexes, while the mean diastolic pressure remained the same in all age groups in females and decreased after the age of 50 years. The prevalence of systolic hypertension among men and women was higher in Manus than in urban Port Moresby and, among the female subjects, Manus had the highest at 31%, while Central recorded the lowest for both males and females. The mean systolic blood pressures of betelnut chewers in Port Moresby, Manus and Central were lower (by 1-9 mmHg) but higher--in males only--in Strickland. The mean diastolic blood pressures of betelnut chewers were lower in all study sites. Both high BMI (overweight and obesity) and older age were significantly associated with high systolic blood pressure but betelnut chewing was significantly associated with lower mean SBP (p < 0.001), a protective effect against systolic hypertension. PMID- 18389972 TI - Thirty years of the Paediatric Standard Treatment Book. AB - The publication of the 8th edition of the Paediatric Standard Treatment Book 30 years after the first edition was introduced in 1975 provided an opportunity to examine the changes in the book's content and composition that have occurred over time. A detailed analysis of all editions of the book was made. The 8th edition is bigger, contains more clinical and guidance topics, and is undoubtedly more complex than the first. Health workers of different levels of training undoubtedly value the book, but there is evidence that it is frequently not used appropriately. The books form an important historical record of the changes in treatment of various clinical conditions that have been driven by alterations in antimicrobial susceptibility and by the emergence of evidence for efficacy. The current book is intricately linked with the Integrated Management of Childhood Illness and the treatments it contains are based on best evidence and practicality. Whilst there are challenges in ensuring that the information in the standard treatment book is accessible, practical and up to date, the book will continue to provide the basis for treatment of the common conditions presenting in children in the future. PMID- 18389973 TI - Nutritional study of the 1-4 year old population of the Lower Jimi Valley, Western Highlands Province, Papua New Guinea. AB - Malnutrition is a significant problem in parts of Papua New Guinea, including the Lower Jimi Valley. There is evidence of specific nutritional deficiencies, such as endemic cretinism, but this study focused on protein-energy malnutrition. It found that, in the 1-4 year old population of three remote villages in the Jimi, only 26% were nutritionally normal, 65% were stunted, 7% were wasted and 2% were stunted and wasted. With respect to weight for age, 16% were 80-100% weight for age, 77% were 60-80% and 7% were < 60%. These figures are considerably higher than those of the National Nutrition Survey of 1982-1983; although the Survey data originated from the entire Jimi District rather than only the villages in this study, these findings suggest that malnutrition is a significant and worsening problem in the Lower Jimi Valley. PMID- 18389974 TI - A qualitative study of beliefs about food relating to child nutrition in the Lower Jimi Valley. AB - A previous study that we carried out confirmed that malnutrition is a problem amongst the young child population in the Lower Jimi Valley. This study begins to explore the cultural and traditional food beliefs in order to target the interventions of health services in reducing childhood malnutrition. The study was undertaken during maternal and child health clinics conducted on a foot patrol from Koinambe Health Centre in the villages of Kompiai, Komengwai, Kupeng, Injim and Kwiama. Semi-structured interviews were carried out with small groups of women to uncover beliefs regarding nutrition and feeding during pregnancy, infancy and childhood. This study has highlighted practices that nutritionally disadvantage young children. The four most significant are late weaning, restriction of foods, infrequency of feeding and the low priority given to children during meal times. It is suggested that it is the use, control and distribution of food, rather than its shortage, that contribute most significantly to the problem of childhood malnutrition in this area. PMID- 18389975 TI - A clinical study of transdermal contraceptive patch in Thai adolescence women. AB - OBJECTIVE: To study cycle control, compliance and safety of a transdermal contraceptive patch in adolescent Thai women. MATERIAL AND METHOD: Fifty-eight healthy women were assigned to receive 3 cycles of contraceptive patch (ethinyl estradiol 20 microg and norelgestromin 150 microg/day). All participants aged 16 20 years were invited to participate from the family planning clinic at King Chulalongkorn Memorial Hospital. Data were collected on adverse effects, perceived advantages and disadvantages, body weight, blood pressure, patch detachments and compliance. Data were analyzed using mean, percentage and student's t-test. RESULTS: The participants' average age was 19.4 years, height 158.8 cm, weight 51.8 kg, BMI 20.8 Kg/m2. The most location of patch application was the abdomen and the most adverse event was breast tenderness (31.0%) followed by application site reaction, nausea vomiting and headache respectively. The breast symptom was mild in severity. The participants reported decrease in dysmenorrhea and shorter duration of bleeding. There were no significant changes in body weight and blood pressure. The improvement of their facial acne was reported. There were no pregnancies during use and the adhesion of the contraceptive patch is excellent. Partial patch detachment was reported in only 6.9%. No completed patch detachment was found. CONCLUSION: The present study found an overall positive impression of a new transdermal contraceptive patch. The good compliance and few side effects were demonstrated. The adhesive patch contraceptive was excellent. PMID- 18389976 TI - Characteristics of pregnancy with human immuno-deficiency virus (HIV) and perinatal transmission in Nakornping Hospital. AB - BACKGROUND: The HIV infected pregnancy delivered at Nakornping Hospital was common. To reduce and prevent HIV infection in pregnancy and perinatal transmission, the understanding of characteristics of HIV pregnancy and neonatal infective outcome were needed for proper strategy and policy making. OBJECTIVE: To study the characteristics of HIV pregnancy and neonatal infective outcome in a hospital in the northern part of Thailand. MATERIAL AND METHOD: This retrospective descriptive study was conducted at the Department of Obstetrics and Gynecology Unit, Nakornping Hospital, Chiang Mai Province, Thailand. Medical records of HIV infected pregnancy from the labor unit during October 2002 and September 2005 was scrutinized. The relevant data of the characteristics of HIV infected mother, pregnancy and neonatal infective outcome were collected and analyzed. RESULTS: There were 172 HIV infected mothers among 7,872 parturients (2.2%). The mean age was 28.2 years (range 14-44 years). Most of the cases were between 25-29 years (40%). 45 pregnancies (26%) delivered before 37 weeks. About half (50.6%) delivered via cesarean section. 22% of their newborn weighted below 2,500 grams. With antiviral regimen of Navirapine and Zidovudine for both mothers and their neonate the overall perinatal transmission rate was 4%. In mothers having ANC group the transmission rate was 3.2% compared to 11.7% in no ANC group. (X2 = 1.092 p = 0.296 Cl 0.04-1.4) RR of ANC group = 0.274 compare to no ANC. CONCLUSION: Many of HIV infected mothers were in the young age group. High preterm labor rate was observed. The no ANC group had about 4 folds infective neonate compared to the ANC group. No antiviral drug during pregnancy in no ANC group may be a factor. This information was vital for strategic ANC planning to prevent and reduce this problem. PMID- 18389977 TI - Effect of audit and feedback on the accuracy of diagnosis summary for gynecological conditions in Songklanagarind Hospital, Songkhla Province, Thailand. AB - OBJECTIVE: To assess the effect of an audit and feedback intervention on the accuracy of the diagnosis summary in the medical records of gynecologic patients in Songklanagarind Hospital. MATERIAL AND METHOD: An audit was conducted on the medical records of 468 gynecologic patients that visited between January 2006 and March 2007. They were divided into non-malignant and malignant conditions both before and after the planned intervention. RESULTS: The accuracy of the summary of the diagnosis for non-malignant conditions was better than malignant conditions. The correction to the principle diagnosis in medical records of patients with malignant conditions increased significantly from 16.7% to 42.9% (p < 0.001) after the audit and feedback. In medical records of non-malignant conditions, corrections to the principle diagnosis and any complications also significantly increased from 72.0% to 82.0% (p = 0.01) and 76.0% to 90.7% (p = 0.002), respectively. CONCLUSION: Audit and feedback is effective for improving the accuracy of the diagnosis summary for gynecological conditions but malignant conditions need more improvement strategies. PMID- 18389978 TI - Dietary calcium intake among rural Thais in Northeastern Thailand. AB - OBJECTIVES: The present study examined the amount and relative contribution of calcium from the habitual diet among rural Thais. MATERIAL AND METHOD: Calcium intake was assessed using 3-day food records and interviewer-administered quantitative food-frequency questionnaire, containing 73 food items. RESULTS: The authors recruited 436 healthy participants (181 men and 255 women), between 20 and 85 years of age. Averaged daily calcium intake among men and women were 378.6 and 265.6 mg, respectively. Sixty-seven percent of men and eighty seven percent of women had less dietary calcium intake than half of the recommended level (< 400 mg/day) whereas only 6 and 3% had an intake more than 800 mg/day. The major food sources of dietary calcium was glutinous rice (32 percent) followed by small animals with edible bones (31 percent) and fresh and fermented fish (20 percent). Dairy products and vegetables constituted only 8 and 5% of dietary calcium, respectively. CONCLUSION: The habitual diet among rural Northeast Thais does not meet the recommended calcium intake level. To promote more consumption of dairy products and locally-available calcium-rich foods would be beneficial to prevent osteoporosis among this population. PMID- 18389979 TI - Incidence and risk factors of nevirapine-associated severe hepatitis among HIV infected patients with CD4 cell counts less than 250 cells/microL. AB - OBJECTIVES: To determine incidence and risk factors of nevirapine (NVP) associated severe hepatitis that led to NVP discontinuation among HIV-infected patients with CD4 < 250 cells/microL. MATERIAL AND METHOD: A retrospective cohort study was conducted among antiretroviral-naive HIV-infected patients who had baseline CD4 < 250 cells/microL and were initiated NVP-based antiretroviral therapy (ART) between January 2003 and October 2005. All patients were categorized to group A: occurred clinical hepatitis and group B: did not occur clinical hepatitis. All were followed until 6 months after ART. RESULTS: There were 910 patients with a mean age of 35.4 years, 57% were males and median (IQR) CD4 cell count was 27 (9-80) cells/microL; contributing 5,006 person-months of observations. Ten (1.1%) patients were in group A and 900 (98.9%) patients were in group B. Incidence of clinical hepatitis was 2 per 1,000 person-months. Probabilities of clinical hepatitis at 0.5, 1, 2, 3 and 6 months after ART were 0.2%, 0.5%, 0.7%, 0.8% and 1.1%, respectively. By Cox regression analysis, baseline AST > or = 1.5 times of upper limit was associated with higher incidence of clinical hepatitis (p = 0.019, HR = 5.83, 95% CI = 1.33-25.51). CONCLUSION: Incidence of NVP-associated severe hepatitis that lead to NVP discontinuation among HIV-infected patients with baseline CD4 < 250 cells/microL is low. The higher baseline AST is also associated with a higher risk of severe hepatitis. PMID- 18389980 TI - Pulmonary arterial hypertension in Thai patients with systemic sclerosis. AB - OBJECTIVE: Study the clinical features of pulmonary arterial hypertension (PAH) in Thai patients with systemic sclerosis (SSc), and compare these features between those with limited (lc) SSc and diffuse (dc) SSc. MATERIAL AND METHOD: The medical records of SSc patients attending the Division of Rheumatology, Chiang Mai University were reviewed. PAH was defined by pulmonary arterial systolic pressure (PASP) > 35 mmHg, determined by Doppler echocardiography. RESULTS: Among 275 patients with SSc, 66 had Doppler echocardiography measurement. Thirty-nine patients (59.1%) had PAH. Among the PAH-SSc patients, 36 (92.3%) presented with dyspnea on exertion, and 37 (94.8%) were in a New York Heart Association functional class of II and III. Twenty-four of 39 patients (61.5%) had interstitial lung disease. Diffuse SSc patients had a significantly higher proportion of males, and shorter disease duration between SSc and PAH diagnosis than lcSSc patients. CONCLUSION: PAH was not uncommon in Thai patients with SSc. Interstitial lung disease might have been the cause associated with over half of these cases. Annual routine Doppler echocardiography screening for PAH in patients with SSc may detect preclinical PAH, and lead to early management and improved functional outcome. PMID- 18389981 TI - Estimation of the proper length of nasotracheal intubation by Chula formula. AB - OBJECTIVES: To assess whether Chula formula can predict the proper length of nasotracheal intubation. MATERIAL AND METHOD: This was a prospective study of 100 patients (50 males and 50 females) who underwent general anesthesia with nasotracheal intubation (number 8.0 for male and 7.0 for female) between December 2005 and March 2007. The Chula formula was used to calculate the length of endotracheal tube (ETT) at the right external naris = 9 + (body height/10) cm. After intubation, a fiberoptic bronchoscope was inserted into the ETT to measure the distance from the tip of ETT to the carina and the distance from the upper border of the cuff of ETT to the vocal cords. RESULTS: The mean length of nasotracheal tube at the right external naris calculated by Chula formula was 25.4 cm in males and 24.4 cm in females. The mean distance from the tip of ETT to the carina was 3.9 cm in males and 3.1 cm in females. The mean distance from the upper border of the cuff of ETT to the vocal cords was 2.6 cm in males and 3.0 cm in females. Ninety-three patients with upper border of the cuff of ETT had it placed at least 2 cm below the vocal cords, and in seven patients, the distance from the tip of ETT to the carina was found to be 1 to 1.9 cm. In every patient, it was found that the tip of ETT was placed at least 2 cm cephalad to the carina. CONCLUSION: In the present study, the Chula formula could be used to predict the proper length of nasotracheal intubation in 93 of 100 patients. PMID- 18389982 TI - Incidence and risk factors of hypotension and bradycardia after spinal anesthesia for cesarean section. AB - BACKGROUND: Hypotension or bradycardia after spinal anesthesia for cesarean section remain common and are serious complications. The current study evaluated factors associated to the incidences of hypotension or bradycardia in this context. MATERIAL AND METHOD: A prospective cross sectional study from November 1, 2004 to July 31, 2005 was conducted on 722 parturients undergoing cesarean section under spinal anesthesia. T-test and Chi-square test were used in univariate analysis to compare continuous data and categorical data respectively. Multivariate logistic regression was performed on the variables hypotension (systolic pressure decrease > 30% of baseline value) and bradycardia (heart rate < 60 bpm) p-value < 0.05 was considered significant. RESULTS: Incidence of hypotension and bradycardia were 52.6% and 2.5%. The probability of hypotension increased with estimated blood loss 500-1000 mL (odds ratio [OR] = 1.86; 95% CI 1.30-2.67, p = 0.001), estimated blood loss > 1000 mL (OR = 5.31; 95% CI 1.47 19.19, p = 0.011), and analgesia level > T4 (OR = 1.94; 95% CI 1.18-3.19, p = 0.009). Hypotension occurred despite left uterine displacement (OR = 1.56; 95% CI 1.11-2.19, p = 0.01). Risk factors associated with bradycardia were adding intrathecal morphine 0.2 mg (0.2 mL) (OR = 4.61; 95% CI 1.31-16.19, p = 0.017) to local anesthetics. CONCLUSION: The present results indicated that the incidence of hypotension after spinal anesthesia for cesarean section increased with amount of estimated blood loss > 500 mL and analgesic level > T4. Adding intrathecal morphine 0.2 mg (0.2 mL) to local anesthetics increased incidence of bradycardia. PMID- 18389983 TI - The Thai Anesthesia Incidents Study (THAI study) of ambulatory anesthesia: II. Anesthetic profiles and adverse events. AB - BACKGROUND: There is a continuing trend to have more elective surgery performed on an outpatient basis. OBJECTIVE: To determine anesthetic profiles and adverse events in practice of ambulatory anesthesia for elective surgery in different levels of hospitals across Thailand. MATERIAL AND METHOD: A prospective descriptive study was conducted in 20 hospitals comprising seven university, five regional four general and four district hospitals across Thailand. Consecutive patients undergoing anesthesia for elective surgery were included. The included patients, classified as outpatients, were selected and extracted for summary of the result by using descriptive statistics. RESULTS: The authors reported 7786 outpatients receiving anesthesia for elective surgery. The majority of patients were in ASA class 1 and 2 (96.2%) while the rest were in ASA class 3 (3.8%). Nearly 90% of the ASA class 3 patients were in university hospitals. The majority of patients (83.1%) did not receive premedication. Diazepam was used more frequently (11.5% vs. 0.1%) than other drugs. Noninvasive blood pressure monitoring and pulse oximetry were used in greater than 90%, while electrocardiogram (EKG) was used in 67.2% and end tidal CO2 in only 6.8%. The three most common anesthetic techniques were general anesthesia (including inhalation anesthetics), total intravenous anesthesia (TIVA), and monitored anesthesia care (MAC). Regional anesthesia was performed in 12% of cases. The three most common regional anesthetic techniques were brachial plexus block (7.1%), spinal anesthesia (2.21%), and other peripheral nerve blocks (2.06%). Propofol was the most common drug for induction. Succinylcholine was used for intubation in 8.8%. The three most common nondepolaring neuromuscular blocking agents were atracurium, cisatracurium, and vecuronium. Fentanyl was the most common drug used among opioids. The three most common volatile anesthetics were sevoflurane, halothane, and isolurane. Lidocaine was the most frequently used in 29.76% of cases, while bupivacaine in 7.9% and ropivacaine only in 0.05%. The majority of the events relating to respiratory system were hypoxia or oxygen desaturation (18:10,000), reintubation (2.6:10,000) and difficult intubation (2.6:10,000), pulmonary aspiration (2.6:10,000), and esophageal intubation (1.3:10,000). Other adverse events included awareness (1.3:10,000), suspected myocardial infarction or ischemia (1.3:10,000), and drug error (1.3:10,000). Five patients (0.06%) received unplanned hospital admission. No patients developed cardiac arrest or died. CONCLUSION: The incidence of major adverse events was low in ambulatory anesthesia for elective surgery when compared to the incidence in general surgical population. The majority of the events occurred in the respiratory system. The authors did not find any complications relating to regional anesthesia. Despite a low incidence of adverse events in ambulatory anesthesia, anesthesia personnel who are responsible for ambulatory anesthesia should have adequate knowledge and skills in selection and preparation of the patients. Therefore, a system of preanesthesia evaluation is very important. PMID- 18389984 TI - A retrospective study of bosentan in pulmonary arterial hypertension associated with congenital heart disease. AB - BACKGROUND: Pulmonary Arterial Hypertension (PAH) plays a significant role in morbidity and mortality of patients with congenital heart disease (CHD). Bosentan, a dual endothelin receptor antagonist has been approved for PAH patients with Eisenmenger physiology (EP). The authors retrospectively reviewed the efficacy and safety of bosentan in Thai PAH patients associated with CHD. MATERIAL AND METHOD: The study population was obtained from the databases of the CHD patients at Siriraj Hospital from October 2004 to April 2007 who received 6 months of bosentan treatment. Inclusion criteria are: CHD with Eisenmenger physiology (EP) or those with severe PAH after surgical repair or interventional cardiac catheterization. Clinical characteristics including the 6-- minute walk test (6MWT) distances, oxygen saturation (O2 sat), New York Heart Association (NYHA) functional class, and right ventricular systolic pressure (RVSP) at baseline were compared with those at 1, 3, and 6 months post bosentan treatment. Signs and symptoms of adverse events were also recorded. RESULTS: There were 11 patients from among those who fitted the inclusion criteria and whose records were examined. Their average age was 51.1 +/- 10.1 years old (13-61 years old). Patients were divided into 2 groups; Group A (6 patients) was PAH with EP and Group B (5 patients) was PAH post intervention. In group A, the 6MWT increased from 151 +/- 69 meters to 293 +/- 61 meters (p = 0.001) with the average increase of 38 +/- 61 meters. The 2O sat increased from 83 +/- 12.7% to 91.8 +/- 5.6% (p = 0.038) with an average increase of 1.4 +/- 0.07%. There was no significant change in right ventricular systolic pressure (RVSP). In group B, there was a trend in 6MWT improvement from 274 +/- 69 meters to 312 +/- 38 meters but this was not statistically different. There were improvements in the NYHA functional class in both groups. There was no significant increase in serum aminotransferase at the end of 6 months in each patient. CONCLUSION: There are benefits of bosentan for treatment of severe PAH in CHD, especially in patients with Eisenmenger physiology. Obvious benefits are an improvement of 6MWT and O2 sat. PMID- 18389985 TI - Comparative study between 2 cm limited quadriceps exposure minimal invasive surgery and conventional total knee arthroplasty in quadriceps function: prospective randomized controlled trial. AB - OBJECTIVE: To compare the recovery time and straight leg raising test after total knee replacement between 2 cm limited quadriceps exposure minimally invasive surgery total knee replacement (2cm Quad MIS TKR) and conventional total knee replacement. MATERIAL AND METHOD: The authors conducted a randomized controlled trial in 40 osteoarthritis knee patients in two groups. Total replacement was performed by the same surgeon and the same prosthesis was used in each group. The function of Quadriceps muscle (straight leg raising test) range of motion was evaluated postoperatively. RESULTS: The 40 patients enrolled in the present study followed the complete study. After surgery with 2 cm Quad MIS TKA, quadriceps function recovery time was faster than with conventional total knee replacement. The quadriceps function was evaluated by the straight leg raising test (SLRT) in sitting and lay down position and the time of ambulation. CONCLUSION: Operative treatment with 2 cm Quad MIS TKA improved recovery time in quadriceps function when compared with conventional total knee replacement. PMID- 18389986 TI - Pueraria mirifica, phytoestrogen-induced change in synaptophysin expression via estrogen receptor in rat hippocampal neuron. AB - OBJECTIVE: To examine Pueraria mirifica (Leguminosae) containing-phytoestrogen effect on synaptic density and involvement of estrogen receptor. MATERIAL AND METHOD: The level of synaptophysin, a presynaptic vesicle protein, was measured using Western blot analysis and immunocytochemistry in hippocampal primary cell cultures at 6 days in vitro. RESULTS: P. mirifica and 17beta-estradiol (0.1 microM) treatment for 4 days, but not for 2 days, significantly increased synaptophysin immunoreactivity and level of synaptophysin. P. mirifica up to 60 microg/ml resulted in a dose related increase in the level of synaptophysin immunoreactivity. The classical estrogen receptor antagonist, ICI 182 780, significantly blocked P. mirifica-induced increase in synaptophysin. CONCLUSION: P. mirifica-containing phytoestrogen affects synaptic density by inducing synaptophysin expression via estrogen receptor. PMID- 18389987 TI - A two-year experience of implementing 3 dimensional radiation therapy and intensity-modulated radiation therapy for 925 patients in King Chulalongkorn Memorial Hospital. AB - BACKGROUND AND OBJECTIVE: Three dimensional conformal radiation therapy (3D CRT) and intensity-modulated radiation therapy (IMRT) have been implemented at Department of Therapeutic Radiation and Oncology, King Chulalongkorn Memorial Hospital (KCMH) since July 2005. This is the first study in Thailand to evaluate the pattern of care and utilization of 3D CRT and IMRT for treatment in each individual cancer. MATERIAL AND METHOD: Between July 2005 and July 2007, 925 newly diagnosed cancer patients underwent IMRT or 3D CRT at KCMH. The authors retrospectively reviewed the experience and utilization of 3D CRT and IMRT for each disease site and region. RESULTS: There were 471 males and 454 females. There were 332 patients (35.9%) treated with IMRT. Among the 332 IMRT patients, there were 100, 32 and 27 nasopharyngeal, lung and prostate cancers, respectively. On the contrary, 593 patients (64.1%) were treated with 3D CRT. Among these, breast, cervix and lung cancers were the most common diseases. Except for head and neck as well as genitourinary cancer 3D CRT was still the main technique used in more than 60% of the patients at KCMH. CONCLUSION: 3D CRT and IMRT have been successfully implemented at KCMH for 2 years. Three dimensional conformal radiation therapy was still the main technique used in more than 60% of the patients at KCMH. Prospective studies evaluating tumor control and treatment sequelae are expected. PMID- 18389988 TI - An epidemiologic study of the Thai Stroke Rehabilitation Registry (TSRR): a multi center study. AB - OBJECTIVE: To perform the registry of stroke patients receiving the in-patient comprehensive rehabilitation program at main tertiary hospitals from March to December 2006. MATERIAL AND METHOD: Demographic data including medical history and pathology of stroke were recorded. All subjects received a comprehensive rehabilitation program until they reached their rehabilitation goals or discharge criteria. RESULTS: Three hundred twenty seven patients met the inclusion criteria. The mean age was 62 +/- 12 years, and 59% were males. Most of the patients were married (73.1%), lived in an urban area (62.1%), and had an education level of primary school or lower (58.7%). The median duration from onset to admission for rehabilitation was 24 days. The major medical history was hypertension (74.9%), followed by dyslipidemia (54.4%), diabetes mellitus (26.6%), and ischemic heart disease (18.0%). Fifty-one (15.6%) patients had a history of previous stroke. Cerebral infarction was found in 71.9%, including thrombosis (45.3%), lacuna infarction (15.3%), and emboli (8.0%) and 28.1% had hemorrhagic stroke. On admission, more than half (51.8%) had cognitive impairment and one-third (31.5%) had bowel-bladder problems. Almost all of the patients (99.4%) had family support. Either their spouse or siblings had undertaken the main caregiver role (46.5% and 40.4% respectively). However, more than 80% of the patients were discharged to their own homes or immediate family's house. CONCLUSION: This was the first multi-center registry of inpatient stroke rehabilitation in Thailand. It presented the epidemiologic aspects in order to become national data of stroke patients receiving medical rehabilitation services. PMID- 18389989 TI - Factors associated with vocational reintegration among the Thai lower limb amputees. AB - BACKGROUND: Vocational reintegration is an important goal in the rehabilitation process. The amputees had to take time to adjust themselves to their new condition after lower limb amputation and that might have an impact on their vocation. As yet, there has been no information regarding vocational reintegration among the lower limb amputees in Thailand. OBJECTIVES: To study the rate of vocational reintegration, factors associated with vocational reintegration among the lower limb amputees. MATERIAL AND METHOD: The postal survey with the questionnaires to 1,300 amputees who received the lower limb prostheses between 2001 and 2005 was conducted. The participants were divided into two groups namely the employed and unemployed groups. The Chi-Square and the Independent Sample T Test were used to compare the difference between the two groups. The multiple variables analysis by stepwise logistic regression was used to determine the associated factors. RESULTS: Three hundred and nine questionnaires were completed among 321 returned questionnaires (response rate 24.7%). Two hundred and forty seven males and 62 females with age ranging from 18 82 years old participated. The rate of vocational reintegration was 66.7%. Demographically, the employed group had less diabetes mellitus (p = 0.001), higher educational level (p = 0.004), were younger at the time of amputation (p < 0.001) and etiologies of amputation were blast injury from mines and congenital problems (p = 0.005). Prosthetic use and problems: the employed group used no gait aids (p < 0.001), had satisfactory to good wearing comfort (p = 0.005), wore prostheses longer hours per day (p < 0.001). The factors associated with vocational reintegration were etiologies of amputation from the blast injury from mines and congenital problems (OR 3.3), educational level from secondary school (OR 2.3), at least satisfactory to good wearing comfort (OR 1.16), and younger at the time of amputation (OR 0.97) respectively. CONCLUSION: This information can assist the rehabilitation personnel to encourage the vocational reintegration among the Thai lower limb amputees. PMID- 18389990 TI - Emphysematous pyelonephritis: a case report and review of literatures. AB - Emphysematous pyelonephritis (EPN) is a rare fulminating, gas-forming infection of the renal parenchyma. Diagnosis of EPN is difficult if it is based only on the history, physical examination and laboratory results. But the radiological evidence is valuable for diagnosis. CT scan is the most valuable method for diagnosis. This is a case report of EPN diagnosed by plain film of the abdomen and confirmed by non-contrast CT. The patient underwent nephrectomy and EPN was confirmed by pathological finding. Diagnostic investigation of this condition was reviewed. PMID- 18389991 TI - Including MIR of a primary bone leiomyosarcoma that radiologically mimics a giant cell tumor. AB - The authors present a case of a 42-year-old female who developed a leiomyosarcoma of the right proximal tibia that appeared radiologically similar to a giant cell tumor Histology revealed spindle cells running in whorl-like fashion with focal atypia and low mitotic figures. The immuno-stains revealed positive reactivity for alpha-smooth muscle (SMA), muscle actin and cytokeratin (AE1/AE3). The authors rendered a diagnosis of low-grade leiomyosarcoma of bone. The lesion was considered a primary lesion since the patient did not have other leiomyomatous tumors. The MRI showed hypo- to iso- signal intensity on T1-weighted imaging and heterogeneous intensity on T2-weighted imaging. This was likely due to admixed fibrotic tissue in the lesion. The tumor cells were not positive for Ebstein-Barr virus by in-situ hybridization as seen in leiomyomatous tumors in immunodeficiency patients. PMID- 18389992 TI - Rupture of unilateral twin tubal pregnancy. AB - A 32-year-old Thai woman presented with acute severe lower abdominal pain and 8 weeks of amenorrhea. The patient was hypotension, had anemia and signs of lower abdominal peritonitis. Initial diagnosis was tubal pregnancy with rupture. Intraoperatively, there were hemoperitoneum and two fetuses were found in the pelvis. Final diagnosis of ruptured unilateral twin tubal pregnancy was achieved. A right salpingectomy was done. There was no immediate complication. PMID- 18389993 TI - Support from a close female relative in labour: the ideal maternity nursing intervention in Thailand. AB - Childbirth is a significant event in the lives of women and their families. It is a critical time in the human development that transforms women into mothers. Women remember their childbirth for the rest of their lives. Thus, the quality of support that women receive during labour and delivery is important and nurses need to be concerned. Previously, women were delivered at homes; they received emotional support from female relatives. Now women give birth in hospitals; they are separated from their families. Although nurses are adept at providing physical and emotional support, they may have to care for several other women. Nurses sometimes may give support to an individual woman a low priority because they have various clinical responsibilities and paper work. It may create womens' experience emotional loneliness and deal with labour pain and in unfamiliar environment alone. These situations can contribute negative effects on childbirth outcomes. Having a close female relative to support a woman in labour can reduce maternal stress and anxiety and improve childbirth outcomes. The present article provides guidelines for including a close female relative on the labour unit to support a woman during labour and delivery. PMID- 18389994 TI - Burden of osteoporosis in Thailand. PMID- 18389995 TI - Science and society: a stem cell technology model. AB - Stem cell technology has been recognized as an emerging technology that could transform current supportive approach toward curing many chronic disorders and degenerative conditions. Regenerative medicine is the promising area of medical practice in the coming decade. However, stem cell technology also brings up controversial issues from the bioethical perspective such as the destruction of human embryos to derive embryonic stem cells or putting the egg donors at risk when retrieving oocytes used in somatic cell nuclear transfer technique. Recently, scientists have discovered a novel method to derive human embryonic stem cell-like cells (iPS; induced pluripotent stem cells) from human skin cells. This innovative approach would not only be a breakthrough discovery to advance the knowledge of stem cell research and the landmark for future stem cell-based therapy but will also provide viable solutions for social concerns on bioethical issues. PMID- 18389996 TI - The chemical and pharmacological importance of morphine analogues. AB - The object of this review is to summarize the efforts which resulted in the discovery of therapeutically useful morphine-like drugs. The search for new analgesics can be divided into three stages: (a) search for analgesics with high efficacy and reduced unwanted side-effects; (b) understanding of structure activity relationships; (c) studies on the mechanism of pain perception and its alleviation by investigation of the pharmacology of opioids. An immense body of literature has been produced on the syntheses of thousands of new compounds which resulted in the development of detailed structure-activity relationships. The physical and psychologic dependence of opioid analgesics also facilitated investigators to solve the problem of the separation of strong analgesia from addiction liability. In the past decades more mixed agonist-antagonist analgesics, pure antagonists devoid of agonist action and potent opioids like the 6,14-ethenomorphinan derivatives were developed. Naloxone, Naltrexone, Buprenorphine and Pentazocine are the outstanding representatives which are introduced into clinical therapy. PMID- 18389997 TI - Characteristics of the myogenic behaviour of arteries of the common European frog (Rana temporaria). AB - Mammalian small arteries exhibit pressure-dependent myogenic behaviour characterised by an active constriction in response to an increased transmural pressure or an active dilatation in response to a decreased transmural pressure. This study aimed to determine whether pressure-dependent myogenic responses are a functional feature of amphibian arteries. Mesenteric and skeletal muscle arteries from the common European frog (Rana temporaria) were cannulated at either end with two fine glass micropipettes in the chamber of an arteriograph. Arterial pressure-diameter relationships (5-40 mmHg) were determined in the presence and absence of Ca2+. All arteries dilated passively with increasing pressure in the absence of Ca2+. In the presence of Ca2+ proximal mesenteric branches and tibial artery branches dilated with increasing transmural pressure but tone (p < 0.05) was evident in both arteries. A clear myogenic response to a step increase or decrease in pressure was observed in small distal arteries (6 of 13 mesenteric and 7 of 10 sciatic branches) resulting in significantly (p < 0.05) narrower diameters in Ca2+ in the range 10-40 mmHg in mesenteric and 20-40 mmHg in sciatic arteries, respectively. The results demonstrate that arteries of an amphibian can generate spontaneous pressure-dependent tone. This is the first study to demonstrate myogenic contractile behaviour in arteries of nonmammalian origin. PMID- 18389998 TI - Effect of caloric restriction on nitric oxide production, ACE activity, and blood pressure regulation in rats. AB - High calorie intake and high weight gain is one of the worldwide health problems particularly in industrial and developed countries. The subjected individuals are at high risk for developing various disorders such as diabetes and particularly cardiovascular problems. It has been well established that life style modification plays an important role in reducing these problems, particularly weight reduction and caloric restriction (CR) as a non-pharmacological approach. This study sought to examine the possible effect of caloric restriction on nitric oxide production, ACE activity and blood pressure regulation in rat. Two groups of rats were selected as the control (C) and the CR group and a with standard and an every other day diet, respectively, for 4 weeks. At the end of study in the CR group systolic blood pressure was significantly decreased compared to controls. The serum NOx was significantly increased compared to the C group. The serum ACE activity was lower in the CR group. Therefore, it may be concluded that CR could reduce blood pressure by elevating NO production and lowering ACE activity. PMID- 18389999 TI - Prolonged melatonin administration in 6-month-old Sprague-Dawley rats: metabolic alterations. AB - The aim of this work was to evaluate the effect of prolonged melatonin administration on chosen metabolic and hormonal variables in male and female Sprague-Dawley rats. Melatonin was administered in tap water (4 microg/ml) daily from the 6th month of age. Rats were fed a standard type of diet ad libitum and were kept in a light regimen L:D--12:12h. The experiment was terminated after 12 weeks of melatonin administration. Melatonin decreased body mass during the whole experiment in females and from the 42nd day of the experiment in males. Relative heart muscle weight in females and absolute/relative thymus weight in males were increased after melatonin administration. Melatonin decreased glycaemia, heart muscle glycogen concentration in females and liver glycogen concentration in both sexes. Serum insulin concentration in males was decreased; serum corticosterone concentration was increased in both males and females. Serum triacylglycerol and heart muscle cholesterol concentration in females were decreased, however in males serum and heart muscle cholesterol concentration was increased. Liver phospholipid concentration in females was decreased and heart muscle phospholipid concentration in males was increased. Melatonin increased malondialdehyde concentration in heart muscle in males and in liver in both sexes. Melatonin induced prominent sex-dependent changes in both carbohydrate and lipid metabolism. PMID- 18390000 TI - Effect of acute maximal exercise on lymphocyte subgroups in type 1 diabetes. AB - The essential therapy of diabetes mellitus includes medical nutrition therapy (MNT), exercise and medical therapy. Exercise, besides its metabolic effects, has positive influence on the immune system, but some forms of exercise may cause trauma for muscle and skeletal systems, they may also support negative effects on the immune system. Nineteen type 1 diabetic patients (mean age 22.1 +/- 2.8 yrs), followed by Diabetes Outpatient Clinic and twenty age matched male control subjects were included into the study, to demonstrate the effects of maximal, acute exercise on the immune system. The exercise test was performed according to Bruce protocol on treadmill. In diabetic subjects, increased CD19 and CD23 expressions were observed before exercise. In both groups (diabetic/control) CD3, CD4 expressions and CD4/CD8 ratio were decreased following the exercise, however expression of natural killer (NK) cells increased. Compared to type 1 diabetic patients healthy subjects had longer acute exercise that caused the increased level of CD8 expression, however type 1 diabetic patients did not show any difference. These results indicate that submaximal aerobic exercise might be recommended for type 1 diabetics without any complications because of its positive reflection on metabolic control and no negative effects on the immune system. PMID- 18390001 TI - Evaluation of the in vitro effect of a Lantana camara extract on the labeling of blood constituents of rats with technetium-99m. AB - Blood constituents labeled with technetium-99m (99mTc) have been used in nuclear medicine procedures and drugs are capable to interfere on this labeling. Lantana camara (lantana) has medicinal properties and it has been used in folk medicine. The aim is to verify the effect of a lantana extract on the labeling of blood constituents with 99mTc. Blood of rats was incubated with extract, stannous chloride and 99mTc, as sodium pertechnetate. Plasma (P) and blood cells (BC) were isolated, also precipitated with trichloroacetic acid and soluble (SF) and insoluble fractions (IF) were separated. The % of radioactivity (%ATI) in these samples was calculated. Samples of labeled BC were washed and the %ATI maintained (%ATI-M) in the BC was determined. The results showed that lantana extract decreased significantly (p < 0.05) in the IF-P from 70.24 +/- 2.59 to 11.95 +/- 3.07. This effect was not observed in the BC and IF-BC. The BC-%ATI-M was significantly (p < 0.05) decreased in all concentrations tested when the BC was washed. This fact was not observed in the control. Substances present on the extract should have redoxi action decreasing the concentration of the stannous ion and this condition could justify the effect on the IF-P. The results about the BC-%ATI-M should indicate a possible effect on the transport of ions through the erythrocyte membrane. PMID- 18390002 TI - Altered insulin sensitivity, insulin secretion and lipid profile in non-diabetic prostate carcinoma. AB - Insulin can influence cancer risk through its effect on cell proliferation, differentiation and apoptosis. Although hyperinsulinemia is considered as a risk factor in the pathogenesis of various cancers, the data related to insulin sensitivity, insulin secretion and lipid profile is lacking in non-diabetic prostate carcinoma cases. The present study was undertaken to evaluate lipid profile parameters and insulin sensitivity and secretion using surrogate markers derived from the measurements of fasting glucose and fasting insulin. The study group comprises 27 prostate carcinoma cases and 27 controls having similar age. Fasting serum insulin, glucose and lipid profile parameters were estimated in both the groups. Insulin sensitivity was assessed by Homeostasis model assessment of insulin sensitivity and Quantitative insulin sensitivity check index. Insulin secretion was assessed by insulinogenic index. Fasting serum insulin, insulinogenic index and LDL-cholesterol were significantly increased (p < 0.05) and HOMA-IS, QUICKI and HDL-cholesterol was significantly decreased (p < 0.05) in carcinoma cases compared to controls. PSA level was significantly associated with fasting insulin (R2 = 0.150, beta = 0.387, p = 0.046) and QUICKI (R2 = 0.173, beta = -0.416, p = 0.031). Fasting insulin was significantly correlated with triglyceride (r = 0.404, p = 0.037) and HDL-cholesterol (r = -0.474, p = 0.013). The present study concludes that hyperinsulinemia associated with reduced insulin sensitivity may play a role in the pathogenesis of prostate carcinoma. PMID- 18390003 TI - Results of nutritional risk screening of patients admitted to hospital. AB - INTRODUCTION: To identify malnutrition, nutritional risk screening and assessment should be performed. A number of nutritional screening and assessment tools have been developed. MATERIALS AND METHODS: Authors monitored the nutritional state of 1266 patients using the MUST (Malnutrition Universal Screening Tool) method. Measurements and questioning were done at admission. The study was cross sectional, presenting a snapshot about nutritional risk of patients. RESULTS: In this study 21% of the patients had a BMI smaller than 20.29% patients admitted to hospital lost at least 5% of their body weight in the 3 to 6 months before the admission. Altogether in this survey 41% of the patients are considered to be endangered concerning malnutrition! There is a statistically visible difference at BMI values between the disease groups. DISCUSSION: If only body weight of patients is measured, the rate of malnutrition is about 20%. Using MUST the number of endangered patients is doubled. In these patients it is more likely that any complication would develop. Since there is no significant relation between the age and the risk group, MUST screening can be carried out on all patients. We intended to emphasize the necessity of nutritional risk assessment and nutritional therapy at all levels of health care. PMID- 18390005 TI - A role of the renal kallikrein-kinin system in the kidney. PMID- 18390004 TI - International society for pathophysiology--its short history. PMID- 18390006 TI - Systematic evidence-based literature review. PMID- 18390007 TI - Background and methodology of the older driver evidence-based systematic literature review. AB - In response to demands of health care and community-based systems, occupational therapy practitioners are eager to provide effective services that are client centered, supported by evidence, and delivered in an efficient and cost-effective manner. Findings from the scientific literature provide a source of legitimacy and authority for informing and guiding practice in all areas, including driving and community mobility with older adults. A systematic review examined studies designed to enhance the driving ability, performance, and safety of older adults. This article presents an overview of the methodology used to develop the focused questions related to the person, vehicle, infrastructure, and policy and community mobility; conduct the literature review; and establish quality control. It includes a summary of the evidence and implications of the review for clinical practice, education, and research to ultimately identify interventions effective in supporting older adult participation and engagement in occupation. PMID- 18390008 TI - Evidence-based and occupational perspective of effective interventions for older clients that remediate or support improved driving performance. AB - To assess the effectiveness of person-related interventions on driving ability in older adults, this literature review was completed as a part of the Evidence Based Literature Review Project of the American Occupational Therapy Association. Nineteen articles were incorporated into the systematic review and include interventions in the following areas: visual, cognitive, and motor; educational; passengers; and medical. The results provide inconclusive evidence for the use of interventions such as the Useful Field of View training, home exercise programs, and passenger interactions. Conclusive evidence shows that older adults respond positively to programs stressing self-awareness of driving skills and that some medical interventions affect the ability to drive. Despite limitations, the studies reviewed provide useful information that deserves further exploration. Reading the literature provides therapists with knowledge that might improve client care. Learning about cutting-edge interventions and educating peers and students about evidence-based interventions may lead to safer community mobility for older adults. PMID- 18390010 TI - Critical review and analysis of the impact of the physical infrastructure on the driving ability, performance, and safety of older adults. AB - Literature on the impact of physical infrastructure on older adult safe driving performance was reviewed in 2005 as part of the American Occupational Therapy Association's Evidence-Based Literature Review Project. Existing guidelines for driving environments, related to changes in visual, cognitive, and psychomotor abilities associated with the aging process (as published in the Highway Design Handbook for Older Drivers and Pedestrians, Federal Highway Administration, 2001), are exhaustive, but the authors made no attempt to critically assess the strength of the study design or level of evidence. In laboratory studies since 1999, the interventions lacked applicability to real-life driving environments. Further investigation of the effectiveness of best practice interventions and how the driving environment can better accommodate the needs of older drivers is needed. Occupational therapy interventions that focus on the occupation of driving and compensation and education strategies that allow older adults to drive safely as long as possible are included. PMID- 18390009 TI - Review of the evidence related to older adult community mobility and driver licensure policies. AB - A systematic review of literature related to the effects of drivers' license policies and community mobility programs on older adult participation was completed as a part of the American Occupational Therapy Association's Evidence Based Literature Review Project. The results revealed that states can achieve a goal of reducing traffic crashes, traffic violations, and traffic-related fatalities through relicensing policies requiring in-person renewal and vision testing as well as driving restrictions. Evidence suggests that failure to consider the continued community mobility needs of older adults can result in increasing fatalities among senior citizens using other modes of community mobility. This evidence-based review is informative for occupational therapy practice; however, it does not reveal the association between policy and community mobility interventions and improved engagement and participation. There is a role for occupational therapy in policy development and community mobility programming to promote engagement in occupation to support participation. PMID- 18390011 TI - Evidence-based perspective on the effect of automobile-related modifications on the driving ability, performance, and safety of older adults. AB - A systematic review of literature related to the impact of modifications made to automobiles on the driving ability, performance, and safety of older adults was completed as a part of the Evidence-Based Literature Review Project of the American Occupational Therapy Association. This review evaluated research on high tech options, such as advanced technology systems (Intelligent Transportation Systems) currently in active development by manufacturers and researchers and the effect of features such as the instrument panel and window tinting. Although the evidence related to Intelligent Transportation Systems is inconclusive, studies have indicated that older adults would use selected technology options. Aftermarket window tinting negatively affects older adults' driving performance, and no evidence demonstrates that hydrophobic window treatment improves driving performance. The implications for occupational therapy practice, research, and education also are discussed. PMID- 18390012 TI - Predicting fitness to drive using the visual recognition slide test (USyd). AB - OBJECTIVE: The authors examined the construct and predictive validity and internal reliability of the Visual Recognition Slide Test developed at the University of Sydney (VRST-USyd). METHOD: A historical cohort study using retrospective descriptive analysis of VRST-USyd scores and on-road driving performance for 838 drivers with impairments was conducted. RESULTS: Rasch analysis provided evidence for the construct validity and internal reliability of the VRST-USyd. Goodness-of-fit statistics for all items were acceptable. The test had high participant and item reliability indexes and separated the participants into four groups with varying levels of skill. Using a cutoff score of 95/164, the sensitivity of the test was 81%, and the specificity was 90%. However, when coupled with clinicians' judgment of participants' awareness of their driving performance during the on-road assessment, this score improved. CONCLUSION: There is evidence for reliability and construct and predictive validity of the VRST USyd. The measurement of awareness requires further research. PMID- 18390013 TI - Pilot study investigating infant vagal reactivity and visual behavior during object perception. AB - OBJECTIVE: The authors examined whether changes in vagal tone were related to infant visual attention during auditory and visual events paired (synchronous) and not paired (asynchronous) in time. They predicted that infants would demonstrate greater visual attention to the synchronous slideshow and that vagal tone would decrease with visual attention. METHOD: Nineteen infants, 3.5 months old, watched computer-generated synchronous or asynchronous slideshows of auditory and visual stimuli. Visual behavior and vagal tone data were collected. Vagal tone reflects physiological responses during attention or exposure to mild stressors. Repeated-measures analysis of variance examined differences in vagal tone across conditions. RESULTS: Visual behavior did not differ between the synchronous and asynchronous slideshow conditions. Vagal tone was significantly lower during the asynchronous slideshow. CONCLUSION: Infants may discriminate synchronous from asynchronous stimuli without changing visual behavior. Implications related to play with toys or objects are discussed. PMID- 18390014 TI - Direct observation as a means of assessing frequency of maladaptive behavior in youths with severe emotional and behavioral disorder. AB - OBJECTIVE: The purpose of this pilot study was to identify the incidence of maladaptive behavior in youths enrolled in community living and support training programs. Direct observation and partial-interval time sampling was used to determine occurrence of behaviors to develop effective strategies for group participation. METHOD: Participants were 30 youths ages 10 to 17 and diagnosed with behavioral disorders. Four observers recorded incidence of eight maladaptive behaviors during three group occupations using direct observation across time and setting, twice per week for 8 weeks. RESULTS: Intraclass correlation coefficients (ICCs) were used to describe the interrater reliability of four observers in recording frequency of maladaptive behaviors. Interrater reliability was high to moderate for the most commonly demonstrated behaviors (ICC = 0.83-0.89). Lower agreement was observed in behaviors that were less frequent, such as violent episodes and sexual inappropriateness (ICC = 0.66-0.68). CONCLUSION: Direct observation across time and setting demonstrated good-to-moderate interrater reliability in identifying frequency of behaviors that may interfere with group participation. Observation provides the opportunity to document occupation-based assessment without having to depend on youths' willingness and ability to answer questions. PMID- 18390015 TI - Descriptive study of occupational therapists' participation in early childhood transitions. AB - OBJECTIVE: The study describes the current role of occupational therapists in the transition of young children with special needs from early intervention to preschool and from preschool to kindergarten. METHOD: Questionnaires were mailed to a random sample of 500 pediatric occupational therapists who are members of the American Occupational Therapy Association's School System Special Interest Section. RESULTS: The response rate was 44.3%, with 173 usable surveys. Approximately 40% of therapists working in the early-intervention-to-preschool transition and 32% of therapists working in the preschool-to-kindergarten transition reported not participating fully in the process. The primary barrier to full participation was "not enough time." Evaluation was identified as the most frequently used strategy for participation. Only 23% of respondents reported receiving specialized transition training. CONCLUSIONS: Although limited by a small sample, the study provides an initial description of occupational therapists' participation in early childhood transitions. PMID- 18390017 TI - Stories we teach by: intersections among faculty biography, student formation, and instructional processes. AB - Becoming a professional who embodies certain dispositions is known as identity formation. Little research has explored what intentions educators hold for student identity formation or how such intentions influence their teaching. Nine faculty members (all female) in an occupation-centered curriculum were interviewed and observed over 8 weeks. Data were analyzed primarily through narrative writing and data matrixes. Ultimately, educators in this study taught who they were; each personified an implicit curriculum, which conveyed the kind of people students were to become and the ways of knowing they were to adopt through the culture, artifacts, rituals, and modes of communication within a program. In the formation of self-defining professionals who embody occupational therapy's ethos, equal attention is needed to the biographical experiences of faculty members, to thoughtful design of the implicit curriculum, and to the ways of knowing portrayed to students every day in the classroom. PMID- 18390016 TI - Revised Knox Preschool Play Scale: interrater agreement and construct validity. AB - OBJECTIVE: Interrater agreement and construct validity of the Revised Knox Preschool Play Scale (RKPPS) were examined. METHOD: Two separately trained raters evaluated 38 typically developing children, ages 36 to 72 months. For each child, the raters observed two 15-min free-play sessions. RESULTS: For the overall play age, the scores of the two raters were within 8 months of each other 86.8% of the time; for the 4 dimension scores, they were within 12 months of each other 91.7% to 100% of the time; and for the 12 category scores, they were within one age level of each other 81.8% to 100% of the time. Construct validity results showed a general match between the children's chronological ages and their overall play age scores. CONCLUSIONS: Findings suggest that two raters can score the RKPPS with some consistency and that scores on this measure progress developmentally, thus supporting its construct validity. PMID- 18390018 TI - Occupational therapy students' metaphors for helping. AB - Metaphors are powerful devices for eliciting images of practice. Exploring the metaphors of occupational therapy students provides educators with insight into students' prior knowledge and the constraints their ideas may present in practice. Metaphorical images of helping held by newly enrolled and Level II fieldwork students were examined. Responses to a structured, open-format questionnaire revealed that the two groups were in agreement about conceptualizations of helping. The findings suggest two overarching themes: (1) the importance of client-centered practice and (2) the inevitability of client autonomy and responsibility. These results imply that educators must prepare students to face the realities of practice: working with unmotivated clients and engaging them in meaningful occupations. Thus, a continued emphasis on client centered practice and the requisite listening skills for a therapeutic alliance are needed. Research should build on the insufficient knowledge of what happens to identity development in the transition from curricula to practice. PMID- 18390019 TI - State of the science on postacute rehabilitation: setting a research agenda and developing an evidence base for practice and public policy--an introduction. AB - Postacute rehabilitation care is a key component of the health care delivery system, yet we know little about the active ingredients of the rehabilitation process that produce the best outcomes. Well-designed research is needed to develop better measures for case-mix adjustment and outcomes of care. To advance research into rehabilitation effectiveness and support the development of evidence-based policies, we must develop new and improve existing measures of patient characteristics, treatment contents, and long-term outcomes. Critical research needs to include --Developing validated measures of rehabilitation interventions and case mix; --Standardizing PAC measures and timing of routine measurement for payment and quality assurance purposes across sites of care; - Examining differences in content and processes of care both within facilities of the same type and across the different types of facilities; --Identifying patient characteristics that vary by region such as rural and urban mix, cultural characteristics, and provider referral patterns; and --Implementing a strategic plan for effectiveness research that is characterized by collaboration among CMS, federal research funders, researchers, and care sites. PMID- 18390020 TI - A randomized controlled trial using patient navigation to increase colonoscopy screening among low-income minorities. AB - Organizational barriers play a key role in colorectal cancer (CRC) screening disparities in low-income minorities. This is a prospective, randomized trial to determine whether a patient navigator (PN) can help overcome the organizational barriers low-income minorities face in trying to obtain screening colonoscopy. Patients of average risk for CRC were referred by their primary care physician for screening colonoscopy. After the PN received the referral, patients were randomly assigned to either receive navigation (PN+) to screening colonoscopy or not receive navigation (PN-). We hypothesized that a PN would increase patient compliance with screening colonoscopy. A total of 21 patients were enrolled in the pilot study (PN+ = 13, PN- = 8); 54% of navigated patients completed screening colonoscopy versus 13% of nonnavigated patients (p = 0.058). Eighty-six percent of navigated patients had an excellent or very good colon prep; however, there was no difference in prep quality between groups ( p = 0.10). One-hundred percent of navigated patients were very satisfied with navigation services. A PN improves compliance with screening colonoscopy in low-income minorities. Larger studies are needed to evaluate what features of navigation are most effective in facilitating completion of screening colonoscopy. PMID- 18390021 TI - Evidence of acute pancreatitis in a patient with radioresistant-differentiated thyroid cancer. AB - Peroxisome proliferator-activated receptor (PPAR) agonists are sometimes used in patients with radioiodine-resistant differentiated thyroid cancers in order to effect further differentiation of the tumor and increase the chance of therapeutic success with subsequent doses of radioiodine. PPAR agonists are reportedly protective of the pancreas and have been proposed as agents that might be useful in the prevention of pancreatitis. In this report, we describe a patient treated with the PPAR agonist rosiglitazone for thyroid cancer who showed imaging abnormalities on PET scan and biochemical evidence of acute pancreatitis. Despite evidence of acute pancreatitis, the etiology of which is unclear, the patient remained asymptomatic. It is speculated that the lack of symptoms in this patient was due to the suppression by rosiglitazone of proinflammatory cytokines. PMID- 18390022 TI - Enrollment of racial and ethnic minorities in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. AB - BACKGROUND: Minority populations in the United States, especially blacks and Hispanics, are generally underrepresented among participants in clinical trials. Here, we report the experience of enrolling ethnic minorities in a large cancer screening trial. METHODS: The Prostate, Colorectal, Lung and Ovarian (PLCO) Cancer Screening Trial is a multicenter randomized trial designed to evaluate the effectiveness of screening for the PLCO cancers. Subjects were recruited at 10 U.S. centers between 1993 and 2001. One screening center had a major special recruitment effort for blacks and another center had a major special recruitment effort for Hispanics. RESULTS: Among almost 155,000 subjects enrolled in PLCO, minority enrollment was as follows: black (5.0%), Hispanic (1.8%) and Asian (3.6%). This compares to an age-eligible population in the combined catchment areas of the PLCO centers that was 14.0% black, 2.9% Hispanic and 5.4% Asian, and an age-eligible population across the U.S. that was 9.5% black, 6.5% Hispanic and 3.0% Asian. About half (45%) of Hispanics were recruited at the center with the special Hispanic recruitment effort. Seventy percent of blacks were recruited at two centers; the one with the major special recruitment effort and a center in Detroit whose catchment area was 20% black among age-eligibles. Blacks, Hispanics and (non-Hispanic) whites were all more highly educated, less likely to currently smoke and more likely to get regular exercise than their counterparts in the general population. CONCLUSION: Significant efforts were made to recruit racial/ ethnic minorities into PLCO, and these efforts resulted in enrollment levels that were comparable to those seen in many recent cancer screening or prevention trials. Blacks and Hispanics were nonetheless underrepresented in PLCO compared to their levels among age-eligibles in the overall U.S. population or in the aggregate PLCO catchment areas. PMID- 18390023 TI - Misestimation of peer tobacco use: understanding disparities in tobacco use. AB - Blacks experience disproportionately elevated rates of tobacco-related morbidity and mortality. Blacks experience delayed smoking initiation relative to other racial/ethnic groups, highlighting the importance of examining smoking correlates occurring in late adolescence/early adulthood. The current study reports data collected as part of an ongoing collaborative effort to assess alcohol and drug use on the campuses of historically black colleges and universities (HBCUs). Two thousand, two-hundred, seventy-seven African-American subjects, aged 20.3 +/- 3.9 (range 18-53), completed the CORE Alcohol and Drug survey and a brief demographic questionnaire. Results indicated that 90% of all subjects overestimated the rate of smoking among their peers. Overestimating was associated with a > 80% increase in the risk of smoking. These data highlight the need to correct misinformation regarding smoking norms among students at some HBCUs. PMID- 18390024 TI - Outcomes of a home-based environmental remediation for urban children with asthma. AB - CONTEXT: Increasing urban asthma prevalence and severity is found among minority, underserved populations. Improving asthma self-management includes home management of complex medical protocols as well as environmental trigger removal. OBJECTIVE: To study the effectiveness of a low-cost approach to improve control of asthma symptoms in an urban population through lay educators who promote a generalized approach to asthma trigger avoidance in the bedrooms of children with asthma. DESIGN AND PATIENTS: Prospective, randomized controlled trial with two arms: historical controls and matched controls for each subject. SUBJECTS: Two hundred-eighty-one patients living in an urban environment, randomized to receive home visits only (n = 128) or home visits with environmental remediation (n = 153). One-hundred-fifteen controls matched according to age, gender and ethnicity did not receive any intervention. INTERVENTIONS: In-home education visits covered asthma physiology, asthma trigger avoidance and asthma management. Environmental remediation was conducted together with the caregiver. OUTCOMES: Primary outcomes include length of hospital stay, number of emergency visits and number of sick visits. Secondary outcomes are symptom frequency, medication management and trigger reduction. RESULTS: Both intervention groups experienced reduction of hospitalizations, emergency room visits, sick visits and asthma symptoms. Both groups showed outcomes significantly superior to the matched control group. Intervention effectively reduced the presence of rodents and carpet in home and increased the use of mattress and pillow covers. CONCLUSIONS: This study shows that low-cost in-home education and environmental remediation improve outcomes for children with asthma. Lay educators can deliver effective asthma-specific education that results in improved asthma control. PMID- 18390025 TI - Sleep duration among black and white Americans: results of the National Health Interview Survey. AB - INTRODUCTION: Epidemiologic studies have shown the importance of habitual sleep duration as an index of health and mortality risks. However, little has been done to ascertain ethnic differences in sleep duration in a national sample. This study compares sleep duration in a sample of black and white participants in the National Health Interview Survey (NHIS). METHOD: Data were collected from 29,818 Americans (age range 18-85 years) who participated in the 2005 NHIS. The NHIS is a cross-sectional household interview survey that uses a multistage area probability design, thus permitting representative sampling of U.S. households. During face-to-face interviews conducted by trained interviewers from the U.S. Census Bureau, respondents provided demographic data and information about physician-diagnosed chronic conditions, estimated habitual sleep duration and functional capacity, and rated their mood. RESULTS: Fisher's exact test results indicated that blacks were less likely than whites to report sleeping 7 hours (23% vs. 30%; chi2 = 94, p < 0.0001). Blacks were more likely to experience both short sleep (< or = 5 hours) (12% vs. 8%, chi2 = 44, p < 0.0001) and long sleep (> or = 9 hours) (11% vs. 9%, chi2 = 23, p < 0.0001). Logistic regression analysis, adjusting for differences in sociodemographic factors, depression, functional capacity and medical illnesses, demonstrated that black ethnicity was a significant predictor of extreme sleep duration (Wald = 46, p < 0.0001; OR = 1.35, 95% CI: 1.24-1.47). DISCUSSION: Independent of several sociodemographic and medical factors, blacks had more prevalent short and long sleep durations, suggesting greater variation in habitual sleep time. Therefore, blacks might be at increased risks of developing medical conditions associated with short and long sleep. PMID- 18390026 TI - Recurrent headache and MRI findings in systemic lupus erythematosus. AB - BACKGROUND: Headache in patients with systemic lupus eryhtematosus (SLE) is considered a common neurological finding, although the relationship is unclear. Another obscure point is the relationship between headache and neuroradiologic findings in these patients. AIM: In this study, we aimed to evaluate the correlation between headache characteristics and intracranial lesions in SLE patients. METHODS AND RESULTS: Forty-eight SLE patients were chosen from those referred to our clinic depending on the American Collage of Rheumatology (ACR) criteria at the same time or after the diagnosis of SLE. Headache classification was done regarding the ICD-II criteria in the patients. Headache severity was assessed by visual analog scale (VAS), and subjects with VAS > or = 4 were included in the study. Patients were divided into two groups according to magnetic resonance imaging (MRI) findings: abnormal MRI (lesion positive) and normal MRI (lesion negative). On MRI, intracranial lesions were detected in 37.5% (n = 18) of the patients, and no lesion was found in 62.5% (n = 30). Headache characteristics were as tension type in 54.1% (n = 26) and migraine like in 39.6% (n = 19) of all patients. Imaging findings were mostly as periventricular and subcortical focal lesions, ranging from 3-22 mm in diameter. A significant correlation was found between abnormal MRI findings with advanced age and prolonged disease duration (p = 0.018, p = 0.016). CONCLUSIONS: As a conclusion, a detailed neurologic evaluation and radiologic investigation, if necessary, should be performed in SLE patients with prolonged disease and advanced age, regardless of headache characteristics. PMID- 18390027 TI - Migraine and Meniere's disease: two different phenomena with frequently observed concomitant occurrences. AB - BACKGROUND: Some variants of migraine could be indistinguishable from Meniere's disease, and this has prompted suggestions of possible association between the two disease entities. AIM AND OBJECTIVES: This study aims at determining the prevalence of migraine among Meniere's patients and a possible linkage between the two diseases in our environment. METHODOLOGY: A 10-year retrospective study of patients diagnosed with Meniere's disease using American Academy of Otorhinolaryngology criteria, including those that had associated migranous symptoms in accordance with the International Headache Society (IHS) diagnostic criteria for migraine, between 1996 and 2005. The prevalence of migraine in the Meniere's patients was also compared with lifetime prevalence recorded from a previous population-based study within the same setting. The statistical difference was tested with a Z nonparametric test (significance at P < 0.05). Epicalc2000 and SPSS 11 were used for the statistical analysis. RESULT: A total of 25 patients representing 0.22% of the 11,463 ear, nose and throat outpatients seen within the study period met the diagnostic criteria for Meniere's disease. There were nine (36%) males and 16 (94%) females. Their ages ranged 27-65 years, mean 45.25 years +/- 11.05. Eight (32%) met IHS criteria for migraine. There is a statistically significant difference between the prevalence of migraine in Meniere's patients and migraine in the overall population (32% vs. 5.3%, P = 0.000). CONCLUSION: The significant preponderance in the prevalence of migraine in Meniere's disease suggests a link between the two diseases. However, more molecular or genetic studies are needed to unveil this phenomenon. PMID- 18390028 TI - Ischemic duodenal ulcer, an unusual presentation of sickle cell disease. AB - Sickle cell disease is caused by molecular abnormalities in the formation of hemoglobin, leading to pain crisis from recurrent vascular occlusion by sickled hemoglobin. Impaired flow in the microvasculature can lead to ischemia, tissue infarction and ulceration. Abdominal pain, a common complaint in sickle cell disease, can be due to an uncommon etiology, ischemic duodenal ulceration. This is due to primary mucosal infarction caused by sickling, leading to poor healing of infarcted areas. Prompt endoscopic and/or urgent surgical intervention should be considered, particularly if anticoagulation is an issue, as proton pump inhibitor use is ineffective in healing this type of ulcer. PMID- 18390029 TI - Cholestasis secondary to panhypopituitarism in an infant. AB - Cholestasis occurring in infancy should be evaluated completely to exclude hepatic as well as endocrine or metabolic causes. A rapid diagnosis should be made to ensure that hepatic and neurologic complications are prevented. We describe a rare case of infant cholestasis due to panhypopituitarism resulting in hypoglycemia and liver biopsy findings consistent with bile duct paucity. Existing bile ducts were noted to be small, and electron microscopy demonstrated diminutive and atrophied biliary cells with a diminished bile duct lumen size. Hypoglycemia and cholestasis resolved with treatment of the underlying panhypopituitarism. Panhypopituitarism should be considered in any infant who presents with cholestasis, hypoglycemia, and other manifestations of pituitary malfunction. Growth hormone deficiency may affect bile duct formation as demonstrated in this patient. PMID- 18390030 TI - The Patient Physician Partnership solutions for a depleted physician workforce. PMID- 18390031 TI - The physiology of adiposity. AB - Normal energy homeostasis requires a balance between fat storage and energy utilization that is guaranteed by regulation of one billion fat cells which arguably constitute the body's largest endocrine unit. Such physiology is required to maintain normal adiposity which if depleted from under- or malnutrition results in lipodystrophy that causes hormonal, reproductive, and developmental abnormalities. Conversely, excess adiposity provides inflammatory secretagogues, particularly from central visceral fat depots that enhance insulin resistance, excessive fatty acids with lipotoxicity and hypertension that escalate atherosclerosis including coronary artery disease. This review describes normal adiposity for maintenance of normal body mass and the roles of adipocyte hormones and adipokines for normal regulation of energy storage and its utilization. Therefore, in this context, the roles of leptin, insulin, adiponectin, and lesser known acylation-stimulating protein, visfatin, and apelin are outlined. Further, adipocyte inflammatory secretagogues are outlined that affect diabetes mellitus 2 with insulin resistance,fatty acid lipotoxicity, dyslipidemia, and hypertension that contribute to the metabolic syndrome. These effects are opposed by adipocyte hormones adiponectin, acylation-stimulating protein, visfatin, and apelin that help maintain normal energy utilization. PMID- 18390032 TI - Non-epileptic seizures: a case report. PMID- 18390033 TI - New volunteer health professionals program to strengthen state's response to emergencies. PMID- 18390034 TI - Finding family. PMID- 18390035 TI - Giving advice on social networking sites. PMID- 18390036 TI - When patients cancel, you're on the line. PMID- 18390037 TI - Use marketing to draw in more patients. PMID- 18390038 TI - Reduce liability risk when treating ... disabled patients. PMID- 18390039 TI - Bond with patients when time is scarce. PMID- 18390040 TI - Why my cash-only practice failed. PMID- 18390041 TI - The best programs for your handheld. PMID- 18390042 TI - 'Payment by results tariff must change to reflect value of nursing'. PMID- 18390043 TI - Delivering change. AB - Lynne Leyshon is the driving force behind major changes to maternity services in Torbay. But hands-on midwifery is still her first love. PMID- 18390044 TI - Learning how to listen. AB - Michael Davidson describes his role in oncology and how he fosters expertise in the nursing team he leads, while Kim Manley analyses his methods. PMID- 18390045 TI - Unhealthy states. AB - A groundbreaking report has spelled out the personal and economic effects of chronic disease in the US. PMID- 18390046 TI - Time for nurses to lead. AB - A new focus on patient care is an opportunity for nursing leaders to take a pivotal role. PMID- 18390047 TI - Every little counts. PMID- 18390048 TI - The time isn't right. PMID- 18390049 TI - Maintaining an airway. AB - Patients who require resuscitation often have airway obstruction, usually secondary to loss of consciousness, however, sometimes airway obstruction may be the primary cause of cardiorespiratory arrest (Nolan et al 2005). Airway obstruction can be subtle and is often undetected. The nurse must be able to recognise and treat airway obstruction to help ensure adequate oxygenation. PMID- 18390050 TI - Nursing management of patients who are nil by mouth. AB - This article examines how to manage patients who are nil by mouth (NBM) and maintain optimal nutritional status. Pre-operative fasting and other reasons why patients might be NBM, methods of administering nutritional support and nursing considerations, such as mouth care, urine output, intravenous therapy and skin assessment are also discussed. The article provides an insight into the psychological effects on patients who are NBM and emphasises the importance of patient education. PMID- 18390051 TI - Nutrition and eating difficulties in hospitalised older adults. AB - This article reviews the relationship between ageing and nutrition. The vulnerability of older adults to nutritional decline may be both a cause and an effect of illness. Undernutrition impairs functional ability and quality of life and markedly increases the costs of care. Since the number of older people in the population is increasing rapidly, this is a matter of considerable concern. The problems that may arise following hospitalisation are discussed, showing the importance of effective nutritional screening following admission. Ways of enhancing food consumption are considered highlighting practical actions that may help to promote intake. Specific eating difficulties that impair intake are examined and suggestions provided as to how they can be overcome. The article shows how, with thought and a structured approach, nurses can ensure that appropriate action is taken to meet the individual nutritional needs of older adults. PMID- 18390053 TI - Application required. PMID- 18390052 TI - Anaphylaxis. PMID- 18390054 TI - Young minds. PMID- 18390055 TI - Speaking volumes. PMID- 18390056 TI - Intrauterine contraceptive device (IUCD): acceptability and effectiveness in a tertiary institution. AB - This was a retrospective study of 2648 clients who accepted intrauterine contraceptive devices (IUCDs) at the University of Benin Teaching Hospital between January 1990 and December 1996 and were analyzed for socio-demographic characteristics, side-effects, complications , reasons for discontinuation in relation to duration of use, age and parity. There were 6178 acceptors of family planning methods giving an IUCD's acceptance rate of 42.9%. The mean age and parity of the acceptors were 31.7 +/- 5.7 years and 4 respectively. Most of the clients were married and had formal education. Of the 859 (32.4%) clients who discontinued the use of method to plan for another pregnancy 799 (93%) were in the age range 20-34 years; while 756 (88%) of them were parity one to four. 294 (49%) out of the 600 (22.7%) continuous users already had 5 to 6 children. The continuation rate at the end of one year and two years respectively were 75.1% (1989) and 61.7% (1633). 3.8% (101) discontinued the use of the method because of heavy bleeding. Removal for pelvic inflammatory disease was 1.7% (44); out of which 0.2% (6) discontinued use in the first 3months of insertion. Expulsion rate was 2.5% (65). The first year expulsion rate was 2.1% (55); out of which 43.6% (24) occurred in the first 3months. Unexpected pregnancy occurred in 0.3% (9). In conclusion the IUCD method of family planning was found to be the most commonly chosen, highly effective with a high continuation rate at one year. The side effects and complications were minimal. PMID- 18390057 TI - Consultation-liaison psychiatry: the past and the present. AB - Consultation-liaison (C-L) psychiatry a subspecialty in psychiatry is referred to as the guardian of the psychological and holistic approach to patient. It has been undergoing rapid changes especially in the developed countries but its practice is strongly affected by stigma and negative attitude by professional colleagues. In this review, it is recommended that C-L psychiatry must not be restricted to conspicuous acute psychiatric disorders alone but must have impact across the whole of medical care. C-L psychiatrists ever than before must not only concentrate on managing psychotic patients but must strive to maintain intellectual and clinical leadership for the psychological system in its entirety. There is yet more to be done in the area of research and campaign against stigmatization. PMID- 18390058 TI - Influence of drug promotion on prescribing habits of doctors in a teaching hospital. AB - This descriptive cross-sectional survey was conducted at University of Ilorin Teaching Hospital to examine the influence of drug promotion by drug companies on the prescription habits of doctors in the hospital. Self-administered questionnaires were used to collect information from 137 doctors selected across all the clinical and laboratory departments using proportionate sampling. Majority (89.0%) of the doctors had attended drug promotion forum and were exposed to 64 different branded drugs within 6 months to this study. Fifty percent of the doctors had prescribed promoted drugs for the first time within 6 months to this study and over two-thirds agreed that drug promotion materials served as incentives to prescribe promoted drugs in preference to their alternatives. More than two-thirds of the doctors did not prescribe in generic names, thus making them susceptible to prescribing promoted branded drugs. Drug promotion by drug companies influence prescription habits of doctors in this teaching hospital. This finding though beneficial to the drug companies may not necessarily be cost-effective and to the benefit of the patients. Further studies and attention on this issue in developing countries is necessary with the ultimate aim of protecting the interest of patients in the face of rising cost of pharmaceuticals. PMID- 18390059 TI - Myelographic abnormalities of spinal cord and nerve roots lesions in conventional myelography. AB - The authors review the myelographic abnormalities of spinal lesions on 90 patients evaluated with conventional or fluoroscopy myelography between 1989 and 1995. The value of Conventional myelography in spinal lesions involving the spinal cord and nerve roots are assessed. The subjects include 49 (45.4%) males and 41 (45.6%) females with a mean age of 38.3 years. Majority of the cases 74(82.2%) had specific neurological symptoms while 16 (17.8%) presented with non specific neurological symptoms. On the whole 78 (86.7%) cases were successfully analysed, in which 58 (74%) cases had abnormal myelograms and 20 (25.5%) had normal myelograms. Extradural lesions was seen in 47 (81%) cases, while 6 (10.3%) cases and 2 (3.5%) cases showed Intramedullary and Intradural-extramedullary lesions respectively. 3 (5.2%) cases showed combined lesions. The most common anatomical site or location of abnormal myelograms was the Thoracic region with 31 (53.4%) cases, 16 (27.6%) cases in the lumbar, 10 (17.2%)and 1 (1.8%) cases in the cervical and sacral regions respectively. Conventional myelography can be successfully used to demonstrate myelographic abnormalities, including identification and determination of the extent of the anatomical location of spinal lesions. PMID- 18390060 TI - Erectile dysfunction in Nigerian hypertensives. AB - Erectile dysfunction (ED) is associated with hypertension and its treatment. It is known to impair and quality of life and compliance to treatment. This study excluded diseases which co-exist with hypertension as well as impair erectile function in their own right. This was to see how much of this problem could be ascribed solely to hypertension or its treatment. Thirty six untreated, newly diagnosed hypertensive males, 21 hypertensive males on thiazides and 37 non hypertensive controls were studied. Co-morbidities which on their own could cause ED were excluded. Erectile function, smoking and alcohol habits were documented. Patient groups were matched for age. Smoking rates were low. There was no significant difference in alcohol use among cases. Both hypertensive groups significantly had more ED than controls. Hypertensives on thiazides tended to have more ED than the untreated newly diagnosed group. ED is common in hypertensives even before treatment and tended to rise with thiazide use. Since it affects intimacy between couples adversely, impacting negatively on treatment outcome; it should be sought in all cases before treatment. Where present, appropriate management steps should be instituted to maintain quality of life and ensure compliance. PMID- 18390061 TI - Extended spectrum Beta-Lactamase-producing Klebsiella pneumoniae septicaemia outbreak in the Neonatal Intensive Care Unit of a tertiary hospital in Nigeria. AB - Between March and August 2002 a cluster of nosocomial septicaemia associated with Extended Spectrum beta lactamase (ESBL)-producing Klebsiella pneumoniae was observed in 11 neonates from the neonatal intensive care unit of a 200-bed tertiary hospital in Abuja, the Federal Capital territory of Nigeria. An investigation was conducted to identify the possible reservoirs and mode of transmission. Infection control measures and epidemiologic surveillance were executed. The environment was investigated by collecting and processing several swab samples for microbiological studies. Antibiogram tests and extended spectrum beta lactamase production test were performed on all K. pneumoniae isolates from both the environment and the patients, and all bacteraemic and environmental isolates of K. pneumoniae compared. A total of 30 K. pneumoniae isolates made up of 17 from the patients and 13 from the environment were analysed. An identical antibiogram was found in 24 isolates, which included all the 17 from the patients and the 7 from the hands of staff, sink and incubator surface in the NICU. Mortality rate from the outbreak was 36.4% and constituted 10.8% of all deaths in the unit in 2002. Overall mortality in the unit for 2002 was 28.9%. The outbreak significantly caused more deaths than usual in the unit. The nosocomial septicaemia was caused by a single ESBL-producing strain of K. pneumnoniae brought into the hospital by a neonate delivered and admitted from an external health institution. Sink and the incubator were also contaminated by the same strain. PMID- 18390062 TI - Reliability of physiological cost index measurements. AB - The reliability of physiological cost index (PCI) measurements under steady state, non-steady state and post-exercise conditions was evaluated. Thirty volunteers (15 male and 15 female) aged 20-30 years participated in the study. None of the volunteers had a history of smoking, respiratory disorder or locomotor abnormalities. Each subject walked at his/her preferred walking speed on an electronic treadmill (Weider TM 2000) and pulse rates (radial pulse) were monitored by the palpatory method. The test was performed twice with an interval of one week between tests. The gender effect on PCI values was evaluated using the independent t-test while the test-retest reliability was determined by the Pearson product-moment correlation method. Results showed no consistent gender difference in PCI. It also revealed a high test-retest reliability (r = 0.843 0.944) for the non-steady state PCI (NSSPCI), the steady state PCI (SSPCI) and post exercises PCI (PEPCI). Analysis of variance showed that the correlation coefficient were significantly different from zero (p < 0.001). It is concluded that PCI is an easy to-use, valid and reliable measure of energy expenditure and it is recommended as a useful tool for physiotherapist in the assessment and evaluation of functional performance. PMID- 18390063 TI - Infant and maternal deaths in rural south west Nigeria: a prospective study. AB - Baseline data on neonatal, infant and maternal deaths including factors associated with infant mortality in a rural community are needed to assess the progress being made towards achieving lower rates in Nigeria. In this community based prospective study, baseline data on births and deaths were collected as they occurred for 6 consecutive years and perinatal risk factors associated with these deaths identified. There were 972 live births in the study period. Maternal mortality ratio (MMR) for the period was 2160 per 100,000 and infant and neonatal mortality rates of 65.8 and 32.9 per 1000 live births were obtained. MMR was highest in mothers aged 40 years and above and lower in mothers 15-34 years. Of infants deaths, 18.8% occurred on the first day of life and 32.8% of deaths occurred within one week of birth. Malaria/fever (23.4%), LBW (17.2%), and Vaccine preventable diseases (neonatal tetanus and measles) (12.5%) were the commonest known causes of infant deaths. Perinatal risk factors for infant deaths included being first birth order (RR = 3.1, 2.1-4.7), birth outside the health care facility (RR = 2.5, 1.4-4.3), no attendant at delivery (RR = 2.5, 1.4-4.4); low weight at birth (RR = 2.46 1.01-5.9) and traditional birth attendants at delivery (RR = 1.7, 1.2-2.6). Babies born to fathers who were between the ages of 25-34 years had borderline protection (RR = 0.76, 0.6-1.01). Delivery and perinatal events have a significant impact on infant survival and more needs to be done to integrate infant survival with maternal survival strategies and this should be done at the primary care level. The community must also be educated and empowered to use the facilities for promotive, preventive and curative care. PMID- 18390064 TI - Non-gonococcal bacterial agents of conjunctivitis and their antibiotic susceptibility patterns in Ilorin, Nigeria. AB - Periodic bacteriological surveillance of purulent ocular discharge is a necessity. To determine the prevalent bacterial agents of purulent conjunctivitis and their antibiotic susceptibility patterns at the University of Ilorin Teaching Hospital (UITH), Ilorin, Nigeria. All bacterial isolates obtained during routine processing of conjunctival swab specimens were studied at the Department of Microbiology UITH between January and December 2004. The isolates were obtained after inoculation of the specimens onto blood, chocolate and MacConkey agar plates, incubated at 37 degrees C for 18-24hours under aerobic conditions except for chocolate agar plates which were kept in candle extinction jar. The isolates were identified using a combination of colony morphology, biochemical and/or serological tests as applicable. Antibiotic sensitivity testing was performed using the modified Kirby Bauer method. One hundred and four conjunctival swabs from 104 patients were analysed and 72 were bacteriologically positive, giving prevalence rate of 69.2%. The positive samples were from 42 males and 30 females with M:F ratio of 1.4:1. The age range was from 5 hours of life to 80 years. Well over half (61.1%) of the swabs were from neonatal age group. The isolated organisms included Staphylococcus aureus 43.1%, Klebsiella pneumoniae 22.2%, Pseudomonas aeruginosa 3.9%, Streptococcus pneumoniae 11.1%, and Escherichia coli 9.7%. Most of the isolates were sensitive to fluoroquinolones while erythromycin has the least bacterial coverage. The most common bacterial agent of purulent conjunctivitis in Ilorin is Staphylococcus aureus and fluoroquinolones have the best coverage for common bacterial agents of purulent conjunctivitis. PMID- 18390065 TI - Novel determination of nabumetone, a cox-2 inhibitor precursor via its 4-carboxyl 2,6-dinitrobenzene diazonium (CDNBD) derived AZO dye. AB - A novel colorimetric determination ofnabumetone in tablets has been developed. The assay is based on chemical derivatization (aromatic ring derivatization technique) using newly developed 4-carboxyl-2,6-dinitrobenzene diazonium (CDNBD) ion as the chromogenic derivatizing reagent and resultant formation of azo dye.Optimization studies established an optimal reaction time of 10 minutes at 30 degrees C after mixing the drug/reagent mixture in a vortex mixer for 10 sec. A new absorption maximum (e(max)) was found at 470 nm, which was selected as analytical wavelength. The assays were linear over 1-6 microg/ml of nabumetone and the optimal reaction required a 2:1 reagent/drug stoichiometric ratio. The developed method has a low limit of detection of 0.39 microg/ml, and is reproducible (1.81% RSD). It has been applied successfully to the assay of nabumetone tablets and is of equivalent accuracy (p > 0.05) with the official (B.P) HPLC method. The new method is simple, has the main advantage of employing a more affordable instrumentation and could find application in routine in process quality control of nabumetone tablets. PMID- 18390066 TI - Health related quality of life and its determinants in Nigerian breast cancer patients. AB - Breast cancer, the commonest malignancy in women, is now the most common cancer in Nigeria, but little is known about the quality of life (QOL) of Nigerian women breast cancer survivors and the predictive factors that may influence their QOL. Health-related QOL (HRQOL) was evaluated in 35 Nigerian women receiving radiotherapy for breast cancer at the University College Hospital, Ibadan by using the European Organization for Research and Treatment of Cancer (EORTC) quality of life questionnaire (QLQ - C30 version 3). Linear regression procedures were used to evaluate the association of HRQOL outcomes with medical and socio demographic factors. Physical functioning (PF), Emotional functioning (EF) and Cognitive functioning (CF) scores were above average (76.9 +/- 20.6, 61.9 +/- 30.3 and 60.0 +/- 32.1 respectively) while Role functioning (RF) and Social functioning (SF) scores were below average (46.2 +/- 36.6 and 40.9 +/- 42.8 respectively). High symptom scale scores were recorded for fatigue, pain and financial difficulties (52.7 +/- 32.8, 59.1 +/- 34.4 and 71.4 +/- 38.8 respectively). Age had no significant influence on any of the functional and symptom scale scores as well as the overall QOL. The overall QOL was significantly related to PF, CF and SF. It was also significantly and inversely related to the scores of fatigue, nausea and vomitng, pain, insomnia and financial difficulty. The findings of this study provide important information on QOL and their correlates among Nigerian women undergoing radiotherapy for breast cancer and are helpful in developing treatment strategies accordingly. PMID- 18390067 TI - Trends in clinical autopsy rate in a Nigerian tertiary hospital. AB - The benefits of autopsy in medical practice are undisputed. In spite of these benefits, autopsy rates worldwide are on a precipitous decline. This study aims to determine trends in autopsy rate in a Nigerian tertiary institution from 1984 2003.A retrospective study of the autopsies conducted between 1984 and 2003. The post mortem registers of the department of Pathology University College Hospital Ibadan were reviewed for records of autopsy over the two decades. A total of 30,899 deaths and 3385 autopsy requests were registered during the period of study, giving average annual deaths of 1626 and average annual autopsies of 178 respectively. Autopsy rates at the hospital declined from an average of 19% in 1984 to 3.6% in 2003. The decline affected all clinical departments and both sexes. The highest autopsy rate of 19.9% was in 1986, while the lowest rate of 3% was recorded in the year 2000. There has been a remarkable decline in hospital autopsy rate in the University College Hospital Ibadan over the last 2 decades. The decline affected both adult and pediatric patients, both sexes and all principal medical units. There is need for active involvement of all stake holders including clinicians, pathologists and family members of deceased patients in strategic measures aimed at reversing this trend. An investigation into the factors responsible for this decline with the view to improving the autopsy services in the hospital is also required. PMID- 18390068 TI - Audit of use of the partograph at the University College Hospital, Ibadan. AB - Study assessed documentation on the partograph and its influence on decision making at the University College Hospital (UCH), Ibadan. Partograph records of parturient during 2004 were retrospectively reviewed. Four hundred and forty-five women had partographic monitoring. High-risk patients were more likely to receive closer (quarter-hourly) monitoring than low-risk women (chi2 = 45.7, p < 0.0001). Documentation was high and not influenced by woman's risk or booking status. Descent of presenting part and liquor status were the least recorded parameters. When tracing crossed the alert line (31.2%) or reached the action line (10.1%), augmentation of labour was more often (but not statistically significant) resorted to than emergency Caesarean section. When tracing crossed the action line however, intervention was significantly more likely to be emergency Caesarean section than augmentation of labour (88.2% vs. 11.8%), chi2 = 5.3, p < 0.05. Intervention for inadequate uterine contractions would more likely be augmentation of labour than emergency Caesarean section (81.4% vs. 18.6%), chi2 = 3.9, p < 0.05. This decision was not significantly influenced by the risk status (chi2 = 0.003, p > 0.05). Outcome of labour was favourable for majority of low and high-risk women and their infants. The partograph is universally employed in monitoring of labour at UCH Ibadan. Its use significantly influences decision making and associated with positive labour outcome among low/high-risk parturient. It is recommended as the sine qua non tool for intra-partum monitoring in all health facilities in Nigeria to reduce maternal complications. PMID- 18390069 TI - Scintimammography screening for recurrent breast cancer in women. AB - Scintimammography (SM) is a nuclear medicine technique in which a tumour seeking radionuclide is used to localize or trace an active or residual tumour in breast tissue. It involves the injection ofa radiopharmaceutical into circulation and obtaining a scan of the breast tissue l0-15 min later. This procedure has been shown to be an accurate diagnostic tool in patients with symptomatic breast lesion with a non-diagnostic mammogram and for whom characterizing the lesion becomes important. This retrospective study was done using the case records of all 72 patients who had presented for scintimammography on follow -up for breast conservation surgery from January to August 2005. The result of scintimammography done using 99mTc tetrofosmin method was compared with mammograms and cytology reports when available. A total number of 52 patients, mean age 49.4 +/- 8.71 years (range 35-67) were analyzed. The commonest presentation was induration at site of surgery. In 43 patients without recurrent disease, scintimammography correctly characterized 42 while 8 out of the 10 patients with recurrent disease confirmed by histology reports were correctly identified The sensitivity of this study was 88.8%, while specificity was 95.4%. The positive predictive value was 80%, while the negative predictive value was 95.3%. Given the non-invasiveness of scintimammography and the high diagnostic index, it is a very useful tool in breast cancer diagnosis and followup. Furthermore, the radiation dose to the patient is lower than a chest radiograph and as such the fear of radiation comparatively is milder. PMID- 18390070 TI - Holoprosencephaly in Ibadan, Nigeria: a report of three cases and review of literature. AB - Holoprosencephaly is a rare congenital malformation resulting from failure of cleavage of midline structures of the forebrain and face. It affects 0.49-1.2 in 10,000 births around the world. The nature of this condition in African children is not well documented in literature. We present a cluster of three cases of holoprosencephaly with varying degrees of facial and intracranial malformations seen within a 6 month period at the University College Hospital, Ibadan, Nigeria. PMID- 18390071 TI - Clinical presentation of retinoblastoma: a tertiary hospital experience. PMID- 18390072 TI - In vitro fertilization is expensive: when should a couple be advised to stop trying with their own gametes and seek other options? Review of three cases. AB - PURPOSE: To describe refractory infertility cases that preserved many failed in vitro fertilization (IVF) cycles to achieve the goal of delivering a baby. METHODS: Case reports with a description of the various approaches and change in strategy that finally led to success. RESULTS: Factors causing repeated failures in these cases included adverse effects of the controlled ovarian hyperstimulation regimen on the uterine environment, failure to realize that the sperm from a male with a low hypo-osmotic swelling test can cause embryo implantation failure unless intracytoplasmic sperm injection is performed, and the discovery that sperm abnormalities rather than exclusively oocyte problems can lead to embryo fragmentation. CONCLUSIONS: It is imperative that infertility specialists individualize cases--especially ones that have failed several cycles- and stop using the same process that has repeatedly failed. They should stop and think if there are some less common but important factors they may have overlooked. Merely recommending donor oocytes without exploring other options is inappropriate. PMID- 18390073 TI - Pregnancy outcome following in vitro fertilization-embryo transfer (IVF-ET) in women of more advanced reproductive age with elevated serum follicle stimulating hormone (FSH) levels. AB - PURPOSE: To present data on the chances of pregnancy following in vitro fertilization embryo transfer, according to day 3 serum FSH and age groups in women > or = age 36. MATERIALS AND METHODS: Data were analyzed according to three age groups (36-39, 40-42, > or = 43) and five serum FSH ranges (< or = 10, 11-12, 13-14, 15-16, > or = 17). RESULTS: No live pregnancies were found in women aged > or = 40 with serum FSH > or = 15 mlU/ml but they were seen in women aged 36-39. Live deliveries were seen in women even > or = 43 with serum FSH 13-14 mlU/ml. CONCLUSIONS: The higher the serum FSH and the greater the age, the lower the chances of successful conception. However, reasonable pregnancy rates are found in women aged > or = 36 with serum FSH > or = 15 mlU/ml and a live delivered pregnancy rate of about 10% can occur even in women aged > or = 43 with mild FSH elevations. PMID- 18390074 TI - Difficulty of embryo-transfer (ET) and pregnancy rate based on the uterocervical angle. AB - When the angle formed by the uterine body and cervical axes (uterocervical angle) was less than 115 degrees, a catheter for embryo transfer could not be smoothly inserted into the uterine body, and so a hard catheter was used, which significantly reduced the pregnancy rate and implantation rate. When the uterocervical angle measured before embryo transfer by ultrasonography is less than 115 degrees, careful preparation, such as catheter selection for embryo transfer and the setting of a longer operation time, is necessary. PMID- 18390075 TI - Attitudes towards contraception in three different populations. AB - OBJECTIVE: To investigate contraceptive behavior of women belonging to three different ethnic and/or socioeconomic populations as well as to evaluate the main sources of information concerning contraception in each population. METHODS: 150 Muslim women living in Germany (group A), 120 Muslim women living in Thrace, Greece (group B) and 140 Christian Orthodox women living in Thrace, Greece were enrolled in the study. Attitudes concerning contraceptive practices were assessed by means of a questionnaire. Demographic and socioeconomic characteristics of each group were compared with the method of contraception used. Statistical analysis was performed using one-way analysis of variance (ANOVA), followed by Turkey's test, chi-square test and multiple logistic regression analysis. RESULTS: The contraceptive pill (41.7%), the condom (35.1%), periodic abstinence (24.4%) and interrupted coitus were the most common methods of contraception. The gynecologist (23.4%), the family consultant (12.0%) and the sexual partner (10.2%) were the most usual sources of information. The use of contraceptive pills was more frequent among Muslims from Germany and Christians from Greece (p < 0.001), while the use of condoms was more frequent among Christians from Greece (p = 0.019). The use of IUDs was more frequent among Muslims from Germany and Greece (p = 0.039). CONCLUSIONS: Our study results reveal that there are behavioral differences between race/ethnic groups and minorities regarding contraceptive practices, probably due to different cultural, socioeconomic and educational factors. PMID- 18390076 TI - Women's health measures in two North Carolina regions sampled from the Basic Automated Birth Yearbook (BABY) datasets: experimental findings, methodological limits and future directions. AB - PURPOSE: To compare selected characteristics in two North Carolina counties to document women's health services at the geographical extremes of the state. METHODS: Using aggregated 2004 data obtained from the North Carolina State Center for Health Statistics, obstetric and perinatal characteristics were experimentally analyzed for the westernmost and easternmost counties in North Carolina (Cherokee and Dare County, respectively). FINDINGS: During the experiment period, 489 infants were delivered in Dare County (population 33,518), while 259 births were recorded in Cherokee County (population 25,289). Prenatal care was established by most women in both counties by the second gestational month. Women in Cherokee County were younger and less educated at delivery than women in Dare County, and smoking prevalence was higher in Cherokee County than in Dare County (31.3% vs 12.9%; p <0.01). Cherokee County infants required assisted ventilation and other medical interventions more often than babies born in Dare County (p <0.01) yet significantly fewer cesarean deliveries were performed in Cherokee County than Dare County (25.5% vs 35.2%; p = 0.04). CONCLUSION: This pilot study showed a significantly higher rate of tobacco use, and lower maternal education level in Cherokee County was associated with a higher incidence of multiple maternal complications and neonatal interventions compared to Dare County. Interestingly, the cesarean delivery rate was lower in Cherokee County despite these factors. We found < 10% of babies born in the study regions required any neonatal intervention. Early and almost universal access to prenatal care did not appear to be a problem at either site. Our preliminary comparison identified important limitations in this government-sponsored dataset that rendered logistic regression analysis methodologically impossible. Changes in process could improve surveillance based on patient-level data and facilitate multivariate analysis. Specific interventions to optimize women's health services form the basis of future experimental research, including larger regional populations. PMID- 18390077 TI - Misoprostol and first trimester pregnancy termination. AB - OBJECTIVE: To investigate the efficacy of vaginal administration of 800 microg misoprostol as a single dose without performing post expulsion systematic curettage in first trimester pregnancy termination. METHOD: 113 women, aged 16 44, who requested first trimester pregnancy termination, received 800 microg of vaginal misoprostol. All examined women were divided into two groups depending on gestation age. The first group included of 67 women with up to nine weeks and the second of 46 with up to 12 weeks of pregnancy. RESULTS: Abortion occurred within 24 hours and was completed in 74.3% of the cases. The mean induction-abortion interval was 5.9 +/- 1.7 hours (median 5.5 hours). Side-effects were experienced by 24 women (21.2%). There was no significant difference between groups in the success rate, induction-abortion interval, number of previous deliveries and side effects. CONCLUSION: Misoprostol is an effective agent for first trimester medical termination. PMID- 18390078 TI - Association study between catechol-O-methyltransferase polymorphisms and uterine leiomyomas in a Japanese population. AB - PURPOSE: To investigate a possible association between uterine leiomyomas and catechol-O-methyltransferase (COMT) polymorphisms in a Japanese population. METHODS: We compared the allele frequencies and genotype distributions of the exon 4 NlaIII restriction site polymorphism (RSP), the P2 promoter HindIII RSP at -1217, and the exon 6 BglI RSP in the COMT gene in 250 leiomyoma cases and 182 controls using polymerase chain reaction-restriction fragment-length polymorphism analysis. RESULTS: No significant differences in allele frequencies and genotype distributions of the exon 4 NlaIII RSP, the P2 promoter HindIII RSP at -1217, and the exon 6 BglI RSP were found between uterine leiomyoma cases and controls. Moreover, no associations were noted between these three polymorphisms in COMT genes and leiomyoma size or a family history of uterine leiomyomas. CONCLUSION: COMT gene polymorphisms are unlikely to be associated with an increased risk of uterine leiomyomas in a Japanese population. PMID- 18390079 TI - Vaginal cysts: a common pathologic entity revisited. AB - PURPOSE: To further study the clinicopathological features of benign vaginal cysts. METHODS: We retrospectively studied all cases of benign vaginal cysts diagnosed in our laboratory over the last decade. Pathological findings were correlated with the clinical records of the patients and histochemistry results. RESULTS: Forty cases of benign vaginal cysts were retrieved. There were 12 cases of mullerian cysts (30.0%), 11 cases of Bartholin's duct cysts (27.5%), ten cases of epidermal inclusion cysts (25.0%), five cases of Gartner's duct cysts (12.5%), one endometrioid cyst (2.5%) and one unclassified cyst (2.5%). Patient age ranged from 20 to 75 years with a mean of 35 years, and a peak incidence between 31-40 years (13 cases, 32.5%). The majority of patients were asymptomatic (31 cases, 77.5%). The cyst type which was more frequently associated with symptoms was Bartholin's duct cyst. Most lesions were located in the left-lateral vaginal wall (13 cases, 32.50%). Mullerian cysts were lined by columnar endocervical-like or cuboidal epithelium, whereas Gartner's duct cysts were all lined by cuboidal epithelium. Epidermal inclusion cysts were lined by stratified non-keratinizing squamous epithelium. Bartholin's duct cysts were lined by transitional, mucin rich columnar or squamous epithelium and were frequently accompanied by inflammation. CONCLUSION: Benign vaginal cysts are in the majority of cases asymptomatic and are often incidentally discovered during gynecological examination for other purposes. The differential diagnosis between Mullerian and Gartner's duct cysts requires histochemical evaluation of epithelial mucin production. The pathogenesis of most types of vaginal cysts remains to be clarified. PMID- 18390080 TI - Frequency of rate of body temperature chart at mid cycle in pregnant women and the subsequent effect on pregnancy. AB - PURPOSE OF INVESTIGATION: To determine if changes in basal body temperature (BBT) during the ovuratory phase are related to subsequent effects on pregnancy. METHODS: BBT records from 216 pregnant women in a spontaneous cycle or a clomiphene citrate cycle during a recent 6-year period were studied. The last day of low phase (LDLP) and the number of days until high phase (NDHP) were determined for all subjects. RESULTS: In the spontaneous cycle group, medium cycle cases were most frequent and long-cycle cases were most frequent in the clomiphene cycle group. The NDHP ranged between one and three days in 82.8% of the subjects in the spontaneous cycle group and in 86.1% of the subjects in the clomiphene cycle group. CONCLUSIONS: Our findings demonstrate the importance of properly evaluating an NDPH of two or even three days in a BBT-based assessment of ovarian function in the ovulatory phase. PMID- 18390081 TI - Clinical symptoms and histopathological findings in subjects with adenomyosis uteri. AB - OBJECTIVE: The purpose of this study was to compare the clinical symptoms and histopathological findings in subjects with adenomyosis uteri. METHOD: Infiltration depth and spread of adenomyotic foci together with clinical symptoms and findings were compared in a total of 103 subjects who had undergone hysterectomy and were diagnosed with adenomyosis uteri through histopathological examinations. RESULTS: The spread of adenomyotic foci in myometrial tissues was observed to significantly increase as the depth of myometrial infiltration increased in subjects with adenomyosis (p < 0.05). It was observed that there was significantly higher myometrial infiltration depth in subjects with dysmenorrhea and severe anemia, and diffuse adenomyotic foci in subjects with menometrorrhagia (p < 0.05). CONCLUSION: Increased infiltration depth and spread of adenomyotic foci in myometrial tissues in subjects with adenomyosis uteri were studied. When clinical symptoms and findings in subjects with adenomyosis, such as dysmenorrhea, anemia and menometrorrhagia are compared with these histopathological findings, infiltration depth and spread of adenomyotic foci appear to determine the clinical severity of adenomyosis. PMID- 18390082 TI - Laparoscopic management of adnexal masses in postmenopause. AB - The study included 64 postmenopausal women with adnexal masses. The selection criteria included menopausal status, an ultrasound scan indicating a benign mass and serum levels of CA-125 below the cutoff (35 U/ml). The results of the study confirm that the removal of a cystic mass in postmenopausal patients with laparoscopic surgery is a more valid and acceptable alternative to traditional surgery. PMID- 18390083 TI - Continuous oral or intramuscular medroxyprogesterone acetate versus the levonorgestrel releasing intrauterine system in the treatment of perimenopausal menorrhagia: a randomized, prospective, controlled clinical trial in female smokers. AB - OBJECTIVE: To compare the efficacy of three progestin regimens in perimenopausal menorrhagia. DESIGN: One hundred thirty-two women with menorrhagia were included in this prospective, randomized, comparative trial. Women were randomized to three groups of 44 in each, either to get a single shot of depot medroxyprogesterone acetate, intramuscularly (Group 1), or medroxyprogesterone acetate in a daily dose of 5 mg orally (Group 2), or the levonorgestrel releasing intrauterine system (LNG-IUS) (Group 3). The Mann-Whitney U-test was applied to compare independent groups. RESULTS: Pictorial blood loss assessment chart (PBAC) score, the duration of bleeding and mean hemoglobin level were improved in all groups. Comparing the groups we noted that for the PBAC, there was no statistically significant difference between groups 1 and 2, while group 3 was superior to both groups 1 and 2 (p < 0.05 and p < 0.05, respectively). Mean duration of menstruation showed no differences among the groups. Hemoglobin levels were no statistically significant differences between groups 1 and 2, while group 3 was superior to both groups 1 and 2 (p < 0.05 and p < 0.05, respectively). CONCLUSION: The efficacies of oral and intramuscular medroxyprogesterone acetate in the treatment of menorrhagia were comparable each other, however, the efficacy of LNG-IUS was superior to both. PMID- 18390084 TI - Cervicovaginal fetal fibronectin (FFN) for prediction of preterm delivery in symptomatic cases: a prospective study. AB - OBJECTIVE: To assess the clinical value of cervicovaginal fetal fibronectin (FFN) in the prediction of preterm delivery (PTD) in women with signs and symptoms of preterm labor (PTL). METHOD: This investigation prospectively studied a cohort of a women with symptoms of PTL, between 24 and 37 weeks' gestation with < 3 cm of cervical dilatation and intact membranes. Cases were evaluated in terms of maternal demographic characteristics like age, body mass index, number of parities, previous PTL history, Bishop scores at admission, gestational age at delivery, mode of delivery, use of tocolytic or steroids, presence of histologic chorioamnionitis, neonatal outcomes and delivery before 34 weeks' gestation as well as within seven days of admission. RESULTS: A total number of 68 cases were included in the study. There were no statistically significant differences between positive and negative FFN groups in terms of maternal characteristics, mode of delivery and adverse neonatal outcomes. However, FFN + cases had higher Bishop scores on admission (3.4 +/- 1.2 vs 2.5 +/- 0.3, p = 0.03) and lower gestational age at delivery (33.4 +/- 3.1 weeks vs 36.8 +/- 2.1 weeks, p = 0.002). Likelihood ratio (LR) for positive results was 1.83 (95% CI: 1.61-2.26) for predicting birth before 34 weeks' gestation, with a corresponding negative LR of 0.62 (95% CI: 0.3-1.2). LR for positive results was 4.34 (95% CI: 3.65-5.12) for predicting birth within seven days of testing, with a corresponding negative LR of 0.3 (95% CI: 0.2-0.5). CONCLUSION: Based on the results of cervicovaginal FFN, positive tests represent an increased likelihood of PTD among women with symptoms of threatened preterm labor. PMID- 18390085 TI - A prospective randomized trial of labor induction with vaginal controlled-release dinoprostone inserts with or without oxytocin and misoprostol+oxytocin. AB - OBJECTIVE: This study was designed in an aim to compare the efficacies of three labor induction methods, dinoprostone (PGE2) vaginal insert with or without concomittant oxytocin and misoprostol (PGE1) combined with oxytocin infusion. METHODS: This was a prospective observational trial of nulliparous women undergoing labor induction from December 2006 to January 2007. Inclusion criteria were: gestational age between 36 to 42 weeks, singleton cephalic presentation of the fetus, intact membrane and unfavorable cervical Bishop score < 6, and absence of spontaneous uterine contractions. Participants were then randomly assigned to preinduction cervical ripening with a dinoprostone vaginal insert (10 mg) administered into the posterior fornix for a total of 12 hours without oxytocin (group I); with oxytocin (group II), and with misoprostol (50 microg) intravaginally in the posterior fornix with repeat dosing at 6-hour intervals with a maximum dose of four with oxytocin (group III). RESULTS: A total number of 106 women met the inclusion criteria without distribution for 19 cases in group I, 44 and 43 cases in groups II and III, respectively. There were no statistically significant differences in terms of the demographic characteristics, indication of labor induction, interval from-induction-to delivery, cardiotocographic abnormalities and neonatal outcomes and mode of deliveries among the three groups (p > 0.05). CONCLUSIONS: Three methods of labor induction were equally efficient in achieving succesful delivery without any maternal and fetal adverse outcomes. PMID- 18390086 TI - Ovarian endometriotic cyst in a patient with Ellis-van Creveld's syndrome: case report. AB - Ellis-van Creveld (EvC) syndrome, or chondroectodermal dysplasia, is a rare genetic disorder associated with chondrodysplasia, ectodermal dysplasia, polydactyly, and congenital cardiac malformations. The disorder is due to an autosomal-recessive mutation mapped to chromosome 4p16. It may occur with different phenotypes. The case of an ovarian endometriotic cyst in a patient suffering from EvC syndrome is reported. PMID- 18390087 TI - Anaesthetic burns. AB - Patients frequently complain of numbness after surgery via the Pfannensteil incision. The two cases in this report demonstrate clear evidence of impaired sensory perception following such an incision. While this is often regarded as a trivial complication, a brief warning to patients should help avoid such superficial burns. PMID- 18390088 TI - Conservative treatment of an early ectopic pregnancy in a cesarean scar with systemic methotrexate--case report. AB - PURPOSE OF INVESTIGATION: Pregnancy implanted in a cesarean scar is rare, and is a life-threatening condition due to high risk of uterine rupture, hemorrhage, hysterectomy, and maternal mortality. CASE REPORT: We describe a 26-year-old woman who presented with five weeks of amenorrhea and a serum hCG level of 10,440 mIU/ml. Transvaginal sonography revealed a gestational sac of 15 x 11 mm containing a yolk sac located in a previous cesarean scar. She was successfully treated conservatively with multi-dose methotrexate. No side-effects were encountered. The serum hCG levels were undetectable in 58 days. The patient had normal menstrual cycles afterwards. CONCLUSIONS: In the view of increasing cesarean rates, healthcare professionals should be aware of the possibility of a scar pregnancy and the potentially life threatening sequelae. Early diagnosis by transvaginal sonography can improve outcome and minimize the need for emergent surgery. Conservative treatment with systemic methotrexate is an effective option in selected patients. PMID- 18390089 TI - Congenital cystic adenomatoid lung malformation: report of two cases and literature review. AB - Congenital cystic adenomatoid malformations of the fetal lung (CCAM) are rare embryonic developmental abnormalities. They are considered as benign hamartomatous or dysplastic lung tumors characterized by overgrowth of the terminal respiratory bronchioles at the expense of the saccular spaces. A minority of cases may not be identified by prenatal imaging techniques and the pulmonary lesions are recognized postnatally. Two cases of congenital cystic adenomatoid malformation of the fetal lung diagnosed in our institution during the last four years are reported. The ultrasonographic and pathologic findings of these cases are discussed. PMID- 18390090 TI - Spontaneous heterotopic pregnancy leading to intrauterine abortion and subsequent ruptured ectopic pregnancy with a betahCG of 125 mlu/ml: a case report. AB - A case of heterotopic pregnancy after spontaneous conception with abortion of the intrauterine pregnancy and subsequent rupture of the ectopic pregnancy is presented. A 34-year-old woman at five weeks of gestation presented with a ruptured ectopic pregnancy after spontaneous abortion of an intrauterine pregnancy with mIU/ml beta3hCG as low as 125 mIU/ml. Close monitoring of betahCG and careful ultrasound examination together with patient consultation are extremely important in the management of heterotopic pregnancy, especially in cases of diagnostic uncertainness. PMID- 18390091 TI - Tubal ectopic pregnancy associated with an extraskeletal chondroma of the fallopian tube: case report. AB - Extraskeletal chondroma is a relatively uncommon benign soft tissue tumor, which usually occurs in the hands and feet. The tumor may also occur around the tendon, synovium, or joint capsule. Rare sites are the tongue, testes and liver. Chondroma of the fallopian tube is extremely rare, with only two reports in the English literature. We present the first reported case of extraskeletal chondroma of the fallopian tube causing transportation impairment of the fertilized ovum in a 32-year-old gravida 1, para 1 woman. PMID- 18390092 TI - Leiomyomatosis with multiple extrauterine pulmonary sites: an unusual case report. AB - The objective of the present study is to present a case of a 48-year-old woman with leiomyomatosis and multiple pulmonary metastases. The classification, pathophysiology, clinical signs, treatment and prognosis of this rare case are discussed. Leiomyomatosis is potentially life-threatening while patients with pulmonary metastases are usually asymptomatic and the condition is incidentally discovered. The treatment of leiomyomatosis is not standardized and many possible variations are under investigation. Still the prognosis is usually excellent. PMID- 18390093 TI - Treatment of canine blastomycotic osteomyelitis. PMID- 18390094 TI - The codes of practice. PMID- 18390095 TI - An ethicist's commentary on not telling the truth. PMID- 18390096 TI - Anthrax in Saskatchewan 2006: an outbreak overview. PMID- 18390097 TI - The effect of bovine viral diarrhea virus infections on health and performance of feedlot cattle. AB - The aim of this study was to investigate the effect of bovine viral diarrhea virus (BVDV) infections (unapparent acute infections and persistent infections) on the overall health and performance of feedlot cattle. Calves from 25 pens (7132 calves) were enrolled in the study. Overall and infectious disease mortality rates were significantly higher (P < 0.05) in pens categorized at arrival as positive for type I BVDV and lower in pens that were positive for type II BVDV than in negative pens. Mortality attributed to BVDV infection or enteritis was significantly more common (P < 0.05) in the pens containing persistently infected (PI) calves than in pens not containing PI calves (non-PI pens). There were no statistically detectable (P > or = 0.05) differences in morbidity, overall mortality, average daily gain, or the dry matter intake to gain ratio between PI and non-PI pens. Although type-I BVDV infections in feedlots appear to contribute to higher mortality rates, the presence of PI calves alone does not appear to have a strong impact on pen-level animal health and feedlot performance. PMID- 18390098 TI - Radiographic detection of thoracic lesions in adult cows: a retrospective study of 42 cases (1995-2002). AB - Medical records of 42 cows that underwent both thoracic radiographic and postmortem examinations within a period of 7 days were reviewed to develop an evaluation grid to interpret bovine thoracic radiographs and to determine the sensitivity and the specificity of thoracic radiographs for detection of thoracic lesions, based on postmortem examination. Most cows (64%) had clinical signs of respiratory disease, whereas 19% showed signs of cardiac problems. The sensitivity and specificity of radiographs for identifying cows with thoracic lesions were 94% and 50%, respectively. In this study, with a prevalence of thoracic lesions of 86%, the positive- and negative-predictive values were 92% and 57%, respectively. This study provides an evaluation grid that allows standardization of the reading of bovine thoracic radiographs and the identification of most thoracic lesions. Bovine thoracic radiographs are useful in detecting thoracic lesions in cows. PMID- 18390099 TI - Accuracy of pregnancy diagnosis in swine by ultrasonography. AB - Early and accurate diagnosis of nonpregnant sows and gilts has the potential to increase reproductive efficiency and the financial income in pig production by reducing non-productive days per sow per year. The objectives of this study were to compare the efficiency of pregnancy diagnosis between Doppler Echo+ and real time ultrasonography (RTU) and to compare the efficiency by using RTU at different days post-mating (days 17 to 24) under commercial conditions. In the 1st study, using crossbreed sows and gilts (n=107), pregnancy diagnoses were done with Doppler Echo+ and then with RTU. Between 28 and 65 days of gestation, Doppler Echo+ had 85% sensitivity and 32% specificity, and efficiency was 73%. In the 2nd study, sows (n=142) were scanned for pregnancy diagnosis between 17 and 24 d post-mating (PD1) and reconfirmed between 38 and 45 days of gestation (PD2). After 21 days of gestation, RTU had over 90% sensitivity and 45% specificity, and 70% efficiency. Accuracy between PD1-farrowing was 75.5% and between PD1 and PD2 was 80.6%. In the 3rd study, sows were diagnosed pregnant by RTU (n=151) at 17 to 24 days of gestation or A-mode ultrasound (n=172) at 28 to 30 days of gestation. There were no significant differences in conception rate (P > 0.09) and farrowing rate (P > 0.67) between both groups. Hence, there was no improvement in fertility and farrowing rate by using RTU instead of A-mode ultrasound under commercial conditions. PMID- 18390100 TI - Use of magnetic resonance imaging in the diagnosis of upper respiratory obstruction in a calf. AB - In a calf with dyspnea, a mass located dorsal to the pharynx was visualized by ultrasonography. Magnetic resonance imaging (MRI) revealed a severe enlargement of the left medial retropharyngeal lymph node, compatible with an abscess. This is the first reported case of MRI use in bovine upper respiratory disease. PMID- 18390101 TI - A possible canine tick-bite reaction to Ixodes muris. AB - An Airedale terrier became acutely ill following attachment of an Ixodes muris tick. Clinical signs waned within hours of tick removal, similar to a pattern previously documented in animals harboring I. muris. This supports the theory that I. muris can induce a noninfectious, severe inflammatory reaction in domestic animals. PMID- 18390102 TI - An unusual form of canine babesiosis. AB - An Akita Inu, living in Belgium, was presented with unusual clinical manifestations of acute babesiosis that included neurological signs and pancytopenia. Diagnosis was made by identifying Babesia canis in the blood smear. Neurological signs resolved after treatment with imidocarb diproprionate. Normalization of hematological abnormalities was gradual over 5 months. PMID- 18390103 TI - Obstructive urolithiasis, unilateral hydronephrosis, and probable nephrolithiasis in a 12-year-old Clydesdale gelding. AB - A 12-year-old Clydesdale gelding was presented for colic and dysuria. Obstructive urolithiasis and chronic renal disease were diagnosed via transurethral endoscopy and percutaneous ultrasonography. Nephroliths, hydronephrosis, and peri-ureteral fibrosis were present. Surgical intervention was declined and the gelding was managed medically with antibiotics and dietary modification. PMID- 18390104 TI - Isolated weaning technology: humane benefits and concerns in the production of pork. AB - The age at which piglets are separated from the sow has been identified as an animal welfare risk in current swine production systems. Animal welfare gains of a minimal disease state must be objectively balanced against the welfare costs of early weaning. Veterinary practitioners should continue to participate in the development and validation of humane animal production methods. PMID- 18390105 TI - Canine lymphosarcoma: overcoming diagnostic obstacles and introduction to the latest diagnostic techniques. PMID- 18390106 TI - Diagnostic ophthalmology. Squamous cell carcinoma. PMID- 18390107 TI - [Medicine in the year 2008: is there light visible at the end of the tunnel!]. PMID- 18390108 TI - [Celiac sprue]. PMID- 18390109 TI - [Atrial fibrillation--a cardiovascular epidemic]. PMID- 18390110 TI - [Conventional treatment of atrial fibrillation]. PMID- 18390111 TI - [The progress in diagnosis and treatment of fungal infections has been considerable over recent years. Yet it is not enough]. PMID- 18390112 TI - [Statin pharmacokinetics]. PMID- 18390113 TI - [Monitoring age-dependent effect of anticoagulation treatment in patients with atrial fibrillation]. AB - Oral anticoagulation treatment with dicumarol preparations (warfarin sodium) is the standard in patients with atrial fibrillation. The effect of treatment depends on many factors, especially in elderly patients. In the study, we assessed the effect of treatment in patients with atrial fibrillation hospitalized in our cardiology ward from 2004 to 2005, in the form of a telephone survey (who controlled the treatment--general practitioner or internist?, the last 2 INR results, complications). INR 2.0-3.5 is considered an efficient therapeutic range. The proportion of permanently correctly anticoagulated patients is approximately 47% across the whole age range, the hypothesis of lower efficiency of treatment in elderly patients does not apply (48% of efficiently anticoagulated patients younger than 75 years vs. 46% of older patients--however, the study does not include polymorbid patients who could not take warfarin at all!) The fact whether a patient is monitored by a general practitioner or an outpatient specialist does not make any difference (49% of anticoagulated patients monitored by a general practitioner vs. 52% of patients monitored by an internist). The percentage of severe complications is relatively low (3.4%). PMID- 18390114 TI - [Positive tests for serum anti-endomysium antibodies (AEA) and anti-jejunal antibodies (JAB) and histopathological diagnosis of celiac disease in children]. AB - Non-invasive examination methods are increasingly important in diagnosing celiac disease. New options for diagnosing celiac disease have been discovered in addition to the established biochemical, hematological and other methods as a result expansive progress ofclinical genetics and immunology. At the same time, detection of circulating auto-antibodies is becoming ever more frequent in clinical practice. As a result, many new, clinically highly heterogeneous cases of the disease have been diagnosed and consequently the prevalence of the disease in both child and adult population has grown. Detection of anti-endomysial antibodies (AEA), characteristic for their high sensitivity and specificity, plays an important role in diagnosing and monitoring celiac disease in pediatric practice. Nevertheless, histopathological diagnosis remains the critical tool for definitive diagnosis of the disease. The article refers to relations between the degree of positivity of AEA and JAB antibodies in the IgA class and the respective grade in the Marsh grading system. The objective of the study was to examine AEA and JAB antibodies and the histological picture of the duodenal mucosa in 20 children and adolescents with celiac disease aged from 2 to 18 years. The authors developed a semiquantitative scale of positivity of both the antibodies, which they compared trying to find a correlation between these and the histopathological picture of the duodenal mucosa. The authors point out the need of timely determination of AEA and t-TG (tissue transglutaminase) in patients whose anamnesis, clinical picture or laboratory results may be indicative of celiac disease. PMID- 18390115 TI - [Assessment of plasmatic cell proliferation and apoptosis indexes in the course of multiple myeloma]. AB - INTRODUCTION: The proliferation and apoptosis indexes are a significant prognostic factor in multiple myeloma (MM). The objective of the study was to assess the indexes in the course of MM from the point of view of the disease activity and potential clinical use. METHOD: A sample of 120 patients was studies, of which 49 at diagnosis, 36 in progression and 35 in remission. The proliferation and apoptosis indexes were assessed according to the achieved treatment response and were compared with subsequent progression in patients in remission. The proliferation potential was measured with the propidium-iodide index (PC-PI/CD138), apoptosis was measured with annexine-V (PC-AI/CD138), assessed by the method of flow cytometry. RESULTS: The sample of49 newly diagnosed patients recorded a decrease in PC-PI (M 2.9-2.1, p < 0.0001) and an increase in PC-AI (M 4.7-8.6, p < 0.0001) at remission. Patients without treatment response showed no difference in PC-PI (p = NS), while PC-AI decreased (M 6.2-3.9, p = 0.004). The sample of36 patients in MM progression with treatment response recorded a decrease in PC-PI (M 2.9-2.2, p < 0.0001) and an increase in PC-AI (M 4.6-8.8, p < 0.019). There was no difference in any of the indicators (p = NS) in cases with zero treatment response. 35 patients in remission with developing progression recorded an increase in PC-PI (M 2.3-2.7, p < 0.0001) and a decrease in PC-AI (M 8.3-4.2, p < 0.0001). CONCLUSION: The study has shown that monitoring PC-PI and PC-AI in the course of different phases of MM is beneficial from the point of view of the assessment of changes in the proliferation activity and the degree of apoptosis of myeloma cells. Monitoring the two parametres allows for timely prediction of disease progression. PMID- 18390116 TI - [Conventional treatment of atrial fibrillation in the Czech Republic managed by outpatient cardiologists. Overview of diagnostic and treatment procedures, pharmacological treatment and hospitalisation]. AB - OBJECTIVE: The aim of the study was to analyse general health data, diagnostic and therapeutic procedures, pharmacological treatment and hospitalization of patients with atrial fibrillation (AF) who did not undergo AF catheter ablation and were in the care of outpatient cardiologists. METHOD: Data concerning 306 patients (of which 94 women, aged 64 +/- 11 years) for the preceding 2 years were acquired through a questionnaire containing a set of standardized questions on a simple form sent out to outpatient cardiologists. RESULTS: AF was paroxysmal, persistent or permanent in 141 (46%), 77 (25%) or 88 (28%) patients, respectively. The higher the age, the lower the proportion of paroxysmal AF and the higher the proportion of the permanent form of AF. AF was asymptomatic in 122 (39%) of patients. The most frequent among cardiovascular diseases was hypertension, detected in 220 patients (72%), IHD was present in 83 patients (27%). The mean LV EF was 55 +/- 11% and was significantly lower in patients with permanent AF than in patients with paroxysmal AF (phi 51 +/- 13% vs. phi 58 +/- 9%, P < 0,001). The mean left atrium transversal diameter was 47 +/- 7 mm and was significantly higher in patients with permanent AF than in those with paroxysmal AF (50 +/- 8 mm vs. 44 +/- 6 mm, P < 0,001). 230 patients (75%) received anticoagulation treatment and 43 patients (14%) received antiaggregation treatment. 274 patients (90%) were taking antiarrhythmic drugs (AA); 93 patients were taking 1, 168 patients 2 and 13 patients 3 AA drugs. 167 patients (55%) underwent electrical cardioversion in 362 procedures, 106 patients (35%) underwent pharmacological cardioversion in 239 procedures. Coronarography was performed in 79 patients (26%) of which 59 (75%) had normal results for coronary arteries. Pacemaker due to concomitant sinus node dysfunction was implanted to 27 patients (9%). Ablation for concomitant atrial flutter of type I was performed in 42 patients (14%). AF and associated conditions caused 250 hospitalisations in 144 patients (47%). The average length of hospitalisation was 4.2 +/- 3.2 days. Cardioembolic event was the cause of hospitalisation of 25 patients (8%) out of 29 hospitalisations with the mean length of hospital stay 8.2 +/- 2.9 days. CONCLUSION: The study has shown, in the first place, very high standards of anticoagulation and antiarrhythmic treatment. It has also shown a relatively frequent indication for coronarography, pacemaker implant for relative sinus node dysfunction or ablation for concomitant atrial flutter of type I, i.e. intervention procedures with limited benefit for AF patients. PMID- 18390117 TI - [The use of galactomannan detection in diagnosing invasive aspergillosis in hemato-oncological patients]. AB - PREMISES AND OBJECTIVES: Timely diagnosis is of critical importance for the prognosis of invasive aspergilosis (IA) patients. Over recent years, IA detection of galactomannan using the ELISA method has assumed growing importance. The objective of the study was to analyse the usability of the method in current clinical practice of a hemato-oncological ward. PATIENTS AND METHODS: From May 2003 to October 2006, blood samples were taken from patients at IA risk to detect galactomannan (GM) in serum using the ELISA method. The patients who underwent the tests were classified by the probability of IA presence on the basis of the results of conventional diagnostic methods and section findings. RESULTS: A total of 11,360 serum samples from 911 adult patients were tested for GM presence. IA (probable/proven) was diagnosed in 42 (4.6%) of them. The rates of sensitivity, specificity, positive and negative predictive value of galactomannan detection for IA diagnosis in our ward were, respectively, 95.2%, 90.0%, 31.5% and 99.7%. The principal causes of the limited positive predictive value of the test were the high percentage of false-positive test results (mainly caused by concomitant administration of some penicillin antibiotics or Plasma-Lyte infusion solution), as well as the fact that a large percentage of patients we examined fell within the group of patients with hematological malignity with a very low prevalence of IA. CONCLUSION: GM detection in serum is associated with high sensitivity and excellent negative predictive value in IA diagnosis in hemato-oncological patients. Knowledge and elimination of possible causes of false-positive results as well as focusing the screening on patients at greatest risk of infection are necessary for an even better exploitation of the test. PMID- 18390118 TI - [Differentiated approach to prescription of physical activity in patients with left ventricular dysfunction]. AB - Correctly indicated physical exercise performed and controlled on a regular basis is an inseparable part of treatment and rehabilitation of patients with left ventricular dysfunction. In order to guarantee the best effect and safety of physical exercise, it is necessary to adopt a differential approach to its prescription to patients with different degrees of functional damage. In addition, a number of conditions should be fulfilled, among which, in the first place, the determination of functional classification of patients used in practice and described in the relevant literature (NYHA, AMA, Goldman, Weber). Physical exercise cannot be differentiated only with respect to the degree of dysfunction; other conditioning factors should be taken into consideration, too, among which the relative contraindication of physical strain, somatic condition, physical exercise anamnesis and others (i.e. sex, age, motivation, etc.), causing a high degree of patient heterogeneity. Also described are additional conditions for differentiation and correct application of physical training, which involve the selection of suitable types of exercise and their energetic demands, adequate intensity, frequency and duration; it is also important to determine the available effective and safe methods, programmes and means of training. The article contains examples of the above conditions, as well as classification of physical exercise into functional classes NYHA I-IV. In conclusion, the authors point out the necessity of differentiation of physical training and of cooperation of the cardiologist with the physiotherapist in its indication, implementation and monitoring. PMID- 18390119 TI - [Statin pharmacokinetics]. AB - Reducing high levels of plasmatic lipoids (LDL-cholesterol and triglycerides) is one of the most important steps in the prevention and treatment of cardiovascular diseases. In the majority of cases, treatment based on lifestyle changes (changes in dietary habits, more physical activity) is not sufficient and pharmacotherapy becomes necessary. Statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, are a well tolerated first-choice drug in patients with dyslipidemia. However, great variability of statin effects has been observed in different patients on the same therapy, and the cause clearly resides in different genetic characteristics of each individual, influencing the effect of therapy. The influence of different genetic variants has been described, but the control of response to hypolipidemic therapy is most likely subject to polygenic control. The analysis of multiple gene combinations may help detect the "hyper-" and "hypo " responders, i.e. individuals with a good response to treatment (allowing for starting with a lower dose of the drug), and those with an insufficient response to treatment (in whom statin shall not be the drug of first choice), or it may help detect the patients who are more likely to develop severe adverse events. Studies with different designs describe that for instance genes (and their variants) for cytochromes, apolipoprotein E and A1 and cholesterol 7alpha hydroxylase may be important genetic determinants of the effect of pharmacological treatment of dyslipidemia and play a role in the individualisation of treatment. PMID- 18390120 TI - [The treatment of Waldenstrom macroglobulinemia and the therapy of diseases caused by monoclonal IgM gammaglobulin]. AB - Waldenstrom macroglobulinemia is defined by the presence of IgM type monoclonal immunoglobulin and histologi cal prove of lymphoplasmocytary lymphoma in the bone marrow. Alkylating cytostatic drugs, chlorambucil or cyclofosfamide in monotherapy have been typically used for its treatment. Similarly to other lymphoproliferative diseases with a low degree of aggressiveness, purine analogues (fludarabine and 2-chlordeoxyadenosine) have a faster treatment response and a higher number of treatment responses, especially if administered in combination with alkylating drugs. The monoclonal antibody rituximab (R) also brings considerable improvement to patients suffering from the disease. It is good for cytopenic patients on monotherapy, and for non-cytopenic patients, in combination with other drugs, both with R-CHOP and with a purine analogue and an alkylating cytostatic drug. In recent years, new drugs for the treatment of this disease have been tested (bortezomib, thalidomid, revlimid and others). High-dose chemotherapy with autologous transplantation has a higher treatment response rate than classical chemotherapy. It is recommended in case of insufficient effect of conventional treatment or early relaps. Young patients with suitable profile and insufficient response to conventional treatment can also qualify for allogenic transplantation; graft response against the disease has been observed in almost all patients who underwent allogenic transplantation. However, treatment is often necessary also in diseases caused by IgM monoclonal immunoglobulin, irrespective of it being formed by a malignant or a benign clone, because the characteristics of IgM monoclonal immunoglobulin may endanger the patient's integrity. The article provides an overview of treatment options for Waldenstrom macroglobulinemia and diseases caused by monoclonal IgM. PMID- 18390121 TI - [Recommendations of Czech Rheumatological Society for the treatment of rheumatoid arthritis. Efficacy and treatment strategies]. AB - Rheumatoid arthritis (RA) is an autoimmune disease of unknown aetiology characterized by presence of chronic symmetric synovitis, which leads to the formation of joint erosions. Generally recommended method for activity assessment of RA is so called Disease Activity Score (DAS). In early RA when low disease activity is present with oligo- or monoarthritis antimalarials are drugs of choice, while sulfasalazine (SAS) is recommended in cases with medium activity without erosions. Initial treatment with methotrexate (MTX) or leflunomide (LEF) should be applied in a very active polyarthritis with a rapid development of erosions. MTX is often combined with other disease modifying drugs (DMARD) and the blockers of tumour necrosis factor alpha (TNF-alpha). LEF is to be administered to the patients in whom the other DMARD are contraindicated or not tolerated. In established RA with oligo- or monoarthritis with permanent low activity SAS is DMARD of choice. In cases with insufficient response and medium activity MTX is used and if it is inefficient LEF or combination of DMARD should be considered. In a very active disease with a rapid evolution of erosions high doses of MTX or LEF are recommended. When extraarticular symptoms of RA are present azathioprine is to be applied and in case of involvement of vital organs cyclophosphamide should be used. When DMARD are failing or contraindicated TNF alpha blockers are to be applied. When one TNF-alpha blocker is inefficient it should by replaced by another one from the same group or another biological should be used. For indication of biologicals the activity limit is DAS28 5.1 and the decrease of DAS28 more than 1.2 is an efficacy criterion. Nonsteroidal antirheumatic drugs are an important part in the management of RA, and also corticosteroids are often of used in oral or parenteral form. To the complex therapy of RA nonpharmacological means are usually implemented--different physical procedures and various surgeries. PMID- 18390122 TI - [Recommended diagnostic and therapeutic approach in arterial hypertension--2007 version. Recommendations of the Czech Hypertension Society]. PMID- 18390123 TI - On medicare in Nigeria again. PMID- 18390124 TI - Cardiac pacemaker treatment of heart block in Enugu a 5-year review. AB - BACKGROUND: Symptomatic heart block is a treatable cardiac cause of death which occurs globally. In Nigeria it is increasingly diagnosed and treated with permanent artificial cardiac pacemaker insertion and pulse generator implantation, sometimes after a period of misdiagnosis and inappropriate treatment. METHODS: Twenty-three patients who were diagnosed with symptomatic heart block and surgically treated with permanent artificial cardiac pacemaker in National Cardiothoracic Centre, Enugu, between April 2001 and March 2006 had their case notes retrospectively reviewed and information entered into a proforma. This was analyzed. Patients diagnosed with symptomatic heart block but not treated with artificial cardiac pacemaker insertion were excluded from the study. There were eight such patients who could not afford the cost of surgical treatment during the period under review. RESULTS: The mean age of the patients was 70 years and the commonest presentation was shortness of breath (100%). Hypertensive heart disease was present in 65% of the patients and a history of chronic chloroquine usage was positive in 73% of the patients. Predominant pretreatment pulse rate was in the range of 30-40 per minute (43%) while 21% of the patients had pulse rate below 30 per minute. These categories of patients commonly had Stoke-Adams syndrome. Sixty-seven per cent of the patients had predominantly systolic hypertension on admission and 16% had hypotension. Third degree heart block was present in 65% of the patients and 89% of all patients needed pre-pacing haemodynamic stabilization with positive inotropic/chronotropic drug(s). Treatment consisted of permanent endocardial pacing in 65% and epicardial pacing in 35% of the patients with equally good response in symptoms, haemodynamic parameters and electrocardiographic features. CONCLUSION: Permanent artificial cardiac pacing is, the reliable treatment of symptomatic heart block and should be included in the National Health Insurance Scheme list. PMID- 18390125 TI - Role of iloprost and bosentan in pulmonary arterial hypertension. AB - BACKGROUND: Pulmonary hypertension is a disorder that is perpetually fatal unless a tentative therapy is instituted. It can be described as a syndrome considering its clinical and pathophysiological manifestations. In this disorder, there is an imbalance within the vascular mediators which possibly arises due to pulmonary endothelial cell injury or dysfunction. Pharmacotherapy in PAH is aimed to reverse the imbalance present among the chemical mediators, offer relief to patients from symptoms and prolong their survival. In addition to other supportive measures, iloprost and bosentan form the cornerstone of treatment. Iloprost, a vasodilator and stable analogue of prostacyclin, confers great benefit through vasodilation, antiproliferative effects and inhibition of platelet aggregation. Bosentan, an oral non-specific endothelin-receptor antagonist with dual activity on both ETA and ETB receptors, has been shown to improve the patient's quality of life on the overall. METHOD: Review of relevant literature was conducted using manual library search and internet articles. The key words employed were pulmonary hypertension, prostacyclin, endothelin-receptor antagonist, hereditary haemorrhagic telangiectasia, iloprost and bosentan. The National Heart, Lung and Blood Institute website was also used in the course of this review. RESULTS: Several studies were able to outline the haemodynamic advantages of iloprost and bosentan in pulmonary arterial hypertension, as evident by improvement in six-minute walk test of patients treated with these agents. CONCLUSION: This review was able to outline the pharmacotherapeutic benefits and role played by inhaled iloprost (in addition to its stable nature and minimal adverse effects) and bosentan in the management of PAH. Several studies have shown that these agents improve the patient's quality of life on the overall considering their favourable effect on pulmonary haemodynamics, symptoms reduction and exercise tolerance. PMID- 18390126 TI - Managing chronic otitis media with cholesteatoma report of 223 patients seen in a 5-year period. AB - BACKGROUND: To assess the clinical features, diagnosis, and treatment of patients having chronic otitis media (COM) with cholesteatoma seen in the ENT/Head and Neck Surgery Clinic at the University Clinical Center, Prishtina, Kosovo. METHOD: A systematic review of the medical records of all patients admitted to the ENT clinic having COM with cholesteatoma between January 1999 and December 2003. RESULTS: During this period, 223 patients suffering from COM with cholesteatoma underwent surgical treatment. There were 123 (55%) male and 100 (45%) female patients. Their mean age was 30.7 years, ranging from youngest patient (7-year old girl) to oldest (73-year-old lady). Thirty-three patients developed complications: 17 patients extracranial complications (EC) and 16 intracranial complications (IC). Leading pathogens in this series were Proteus mirabilis in 52% of cases, Proteus vulgaris in 14% and Staphylococcus aureus in 12%. All patients underwent surgical treatment: 10 patients (4.5%) a canal-wall up (CWU) procedure and 213 (95.5%) a canal-wall down (CWD) procedure. No deaths occurred in this series. CONCLUSION: Because of possibility of developing life-threatening complications, patients with COM with cholestatoma must diagnosed in time and followed as soon as possible to the center that performs ear surgery, as only adequate treatment option for these patients. PMID- 18390127 TI - Booking status and caesarean section outcome in LAUTECH teaching Hospital, Osogbo. AB - INTRODUCTION: Antenatal care is concerned mainly with prevention, early diagnosis and treatment of general medical and pregnancy associated disorders. Quality of care varies across health facilities; the booking status of the women who deliver would have influence on the pregnancy outcome. METHOD: A comparative study of caesarean section outcome among the booked and unbooked mothers delivered at Ladoke Akintola University Teaching Hospital, Osogbo. RESULTS: Mean age of the women was 29.7 +/- 5.9 years. The booked mothers were older than the unbooked mothers. All the maternal deaths were among the unbooked mothers. There were higher rates of preterm birth, neonatal asphyxia and neonatal admissions to intensive care units among the babies delivered by unbooked mothers compared to the booked mothers. CONCLUSION: Unbooked women and their babies are at higher risks of caesarean section complications than the booked mothers. PMID- 18390128 TI - Awareness and acceptability of strategies for preventing mother to child transmission of HIV among antenatal clients in Calabar, Nigeria. AB - BACKGROUND: Mother to child transmission is the major route through which children below the age of 15 years acquire HIV infection. The most effective way to reduce childhood HIV infection is to prevent the infection in mothers and for already infected mothers use appropriate strategies to prevent transmission to their children. This study was conducted to determine the level of awareness and acceptability of strategies for preventing mother to child transmission of HIV. METHOD: Exploratory multi-centric descriptive study involving 400 antenatal attendees in Federal, State and a Private health facility was used. Interviewer administered questionnaire was the tool for data collection. RESULT: Majority of the respondents (94.7%) were aware of transmission of HIV from an infected mother to her child. Respondents were more aware of the use of antiretroviral drugs in pregnancy (63.2%) than they were of avoiding breastfeeding (58.5%) and Cesarean delivery (22.8%) as strategies for preventing mother to child transmission. They were also more likely to accept the use of antiretroviral drugs (78.2%) than they would avoid breastfeeding (69.0%) and accept Cesarean delivery (38.0%) for preventing mother to child transmission of HIV High educational status was significantly associated with a positive attitude to these strategies. CONCLUSION: There is need for more educational programs and social support to bridge the gap between the levels of awareness and acceptability of strategies for preventing mother to child transmission of HIV among the populace. PMID- 18390129 TI - Clandestine abortion in Port Harcourt: users' profile and motivation. AB - BACKGROUND: Most of the information on abortion in Nigeria are based on information from women hospitalized for abortion complications. However a lot of abortions, especially those carried out by medical doctors do not cause enough complications to register in hospital statistics. This study is to explore the profile of women that obtain these relatively safe abortions, and to ascertain their reasons for opting for an abortion. METHODS: A cross sectional study was carried out in five clinics located in various parts of Port Harcourt, Nigeria, known to provide abortion services. Data on all the women that sought abortion in the clinics between January and March 2007 were collected by the attending doctors. RESULTS: A total of 793 women sought abortion in the clinics within the three months study period, a daily average of 1.76. The clients had an average age of 23.73 years (s.d 7.3), and a large proportion were not yet married (72.01%), had no children (67.97%), were still in school or learning a trade (66.08%), and were having their first termination of pregnancy (68.22%). Clients that identified themselves as widowed, divorced or separated made up 17.78% of the total. The commonest reasons cited for the decision to terminate the pregnancy were that the women were not yet married (63.43%), and that they were still students or learning a trade (60.15%). Few women gave contraceptive failure (3.91%) or the fact that they did not want any more children (3.66%) as a reason for the abortion. CONCLUSIONS: Abortion on demand is still very common in Nigeria in spite of legal restrictions. Amending the abortion laws would assist in resolving this paradox. PMID- 18390130 TI - Congenital eye and adnexial anomalies in Kano, a five year review. AB - BACKGROUND: Knowledge of prevalence and types of congenital eye and adnexial anomalies is important as some of these anomalies can lead to childhood blindness. The aim of the study is to determine the types of congenital eye and adnexial anomalies seen in the eye clinic of Aminu Kano Teaching Hospital Kano, Nigeria between the years 2001 to 2005. To determine the proportion of children below the age of 10 years who presented with congenital eye and adnexial anomalies, the proportion of eye surgeries due to such anomalies, and proportion of these anomalies amenable to (surgical) intervention. METHODS: The clinic and theater registers were used to obtain the total number of children less than 10 years of age seen and those that had surgery during the review period. A list of those with congenital eye and adnexial anomalies was compiled. The case folders of patients with such anomalies were retrieved and information on age, sex, type of anomaly, laterality of the condition, and types of surgical intervention offered, was obtained. The data was manually analyzed. RESULTS: There were 4163 children seen in the eye clinic and 268 had surgery during the review period. There were 109 eyes of 69 patients with congenital eye and adnexial anomalies. The prevalence of such anomalies amongst children (<10 years of age) is 1.7% and surgery for congenital eye and adnexial anomalies accounted for 25.7% of eye surgeries in the study population. The male to female ratio is 2.3:1. In 40 patients, the anomaly was bilateral and it was unilateral in 29 patients. The commonest congenital anomalies are buphthalmos in 38%, cataracts in 35%, and naso lachrymal duct obstruction in 14%. Other less frequent anomalies are anophthalmia/micophthalmia, limbal dermoid cysts and aniridia. Eighty six percent of the patients had surgery to correct the anomaly. CONCLUSION: Most of the congenital anomalies seen in our hospital can lead to childhood blindness and vigilance by eye care providers is advised to ensure prompt identification and intervention. PMID- 18390131 TI - Determinants of cigarette smoking among senior secondary school students in a rural community of southwest Nigeria. AB - BACKGROUND: Smoking remains an increasing high-risk behaviour among the youth, despite its harmful effects on health. This study sought to find out the determinants of cigarette smoking among youth of a rural Nigerian community and suggested intervention measures which have potential for the control of smoking among in-school population. METHODS: It was a cross-sectional study carried out among 416 students selected through a multi-stage sampling procedure. Determinants of smoking among the study participants and their knowledge were assessed with the aid of a pre-tested structured questionnaire. RESULTS: The proportions of ever-smokers who could associate cigarette smoking with known health problems were generally low compared to the never-smokers. The mean knowledge score, 4.05 +/- 0.4, obtained by the ever-smokers was also lower than the mean score, 6.41 +/- 0.2, obtained by the never-smokers. This was found statistically significant (p < 0.05). Smoking behaviour was significantly associated with friends (p = 0.00518) and parents (p = 0.002856) who smoke, and with cigarette advertisement (p = 0.032989). CONCLUSION: Low level of knowledge, peer and parental influence as well as exposure to cigarette advertisement played significant roles in the adoption of smoking behaviour by the youth. Anti-smoking education and other prevention strategies targeted at the youth, their parents and the media are desirable. PMID- 18390132 TI - Diets rich in vegetables and physical activity are associated with a decreased risk of pregnancy induced hypertension among rural women from Kimpese, DR Congo. AB - BACKGROUND: To assess whether the frequency pf pregnancy-induced hypertension is low, and vegetables intake and physical activity are protective against pregnancy induced hypertension onset among rural women from Democratic republic of Congo. METHOD: This hospital-based and longitudinal study was carried out within the rural hospital of Kimpese, DR Congo were monitored from January 1st to March 31st 2003 on basis of demographic, diet, physical activity, anthropometry and blood pressure till the onset of types of pregnancy-induced hypertension and delivery. RESULTS: Out of 238 black pregnants, the incidence risk of arterial hypertension was 4.6% (n=11) whose 2.9% with Pre-Eclampsia and 1.7% with transient hypertension. Gestity, parity and birth weight of infants were significantly lower among hypertensive mothers, while positive family history and presence of oedemas were more elevated among hypertensive pregnants. The onset of Pre Eclampsia was higher within vendors and inactive women as well as among vegetarians (3.1%) than women with diet high in meat (9.7%). Pre Eclampsia occurred more (p<0.05) among pregnants with rare daily servings of vegetables (33.3%) than pregnants with 3 or more daily servings of vegetables (3.7%). Physical activity (RR=0.63 CI 95% 0.33 to 0.94) and = 3 daily servings of vegetables (RR=8.8 CI 95% 0.6 to 0.98) were significant (p<0.01) protective factors against Pre Eclampsia. Eclampsia was not observed. CONCLUSION: It is timely to promote diet rich in vegetables overweight reduction and physical activity among pregnants for the prevention of pregnancy-induced hypertension. PMID- 18390133 TI - Experience with tetanus in a tertiary hospital in south east Nigeria. AB - BACKGROUND: Tetanus has for long been an avoidable source of morbidity and mortality particularly in developing countries. The aim of this study was to review the experience with managing tetanus in a regional tertiary hospital in Nigeria. METHODS: A retrospective study of patients who were admitted with a clinical diagnosis of tetanus between January 1999 and December 2003 was done. Relevant data were extracted from the patients'case records. RESULTS: A total of 12 patients were identified within the time frame with a diagnosis of tetanus. There were more male than female patients (ratio 1.4:1) with city dwellers constituting a small majority. The mean age of the patients was 29.8 years. Lower limb injuries accounted for the portal of entry in 75% of cases while one case followed a practice of oral sex. All patients were managed in the general ward and no mortality was recorded despite the presence of dysautonomia in some patients. CONCLUSION: Despite the high rate of morbidity and mortality associated with tetanus in developing countries and despite some regional differences in presentation, careful clinical management even in resource poor countries can result in very good outcomes. PMID- 18390135 TI - Neutropenia and human immunodeficiency virus-1 infection: analysis of 43 cases. AB - BACKGROUND: Neutropenia has been reported in patients with early asymptomatic HIV infection as well as in those with more advance HIV-related immunodeficiency. As with other peripheral cytopenias in the setting of HIV infection, multiple aetiologies may be present either singly or in combination. The study aims to determine the prevalence of Neutropenia and the association with the level of deterioration of CD4+ T lymphocyte subset in antiretroviral naive HIV-1 infected patients. METHOD: Four hundred consecutive HIV-1 infected patients undergoing investigations for pre treatment and staging were recruited over a one year period, at the HIV subspecialty clinic of Ahmadu Bello University Teaching Hospital, Zaria. All the patients were confirmed HIV-1 infected, repeatedly reactive by ELIZA. White cell count was determined by standard manual method and CD4+ T cell enumeration by Dynal (Oslo Norway) manual method. RESULTS: Of the 400 patients studied, the prevalence of neutropenia was 17.5%; 4.5% and 13% amongst the asymptomatic and symptomatic group respectively. There is a signification positive correlation between absolute neutrophil count and CD4 + T lymphocyte count p<0.05. CONCLUSION: This study demonstrates that Neutropenia is relatively common in HIV infected patients and is associated with the severity of the immunological deterioration. PMID- 18390134 TI - Management of motorcycle limb trauma in a teaching hospital in south-western Nigeria. AB - BACKGROUND: To evaluate the management of limb injuries sustained from motorcycle accidents. METHOD: Consecutive 115 patients managed for limb injuries secondary to motorcycle trauma over a one-year period were prospectively studied. RESULTS: There were 115 patients out of which seventy-six (66.1%) sustained injuries to the lower limbs, 25 (21.7% to the upper limbs, while 14 (12.2 %) injured both the upper and the lower limbs together. Seventy-nine (68.7%) patients had fractures of which 24 (30.4%) were open. There was no organized pre-hospital care, a high referral rate and long injury-treatment intervals. Treatment was by closed manipulation and splintage with casts (57.6%), tractions (30.4%), and external fixation (5.4%); 18 patients had elective ORIF. The complication rate for cases was 71.3% while 4 (3.5%) patients died. CONCLUSION: Management of motorcycle injuries is still difficult in developing countries. Attention needs to be paid to provision of organized trauma care to improve outcomes. PMID- 18390136 TI - Outcome of directly observed therapy short course (DOTS) regimen in a rural community of the Nigerian Niger Delta. AB - BACKGROUND: [corrected] The objective of this retrospective study was to evaluate the outcome of directly observed therapy short course (DOTS) application in a Nigerian rural community. METHODS: A retrospective study of all the records of DOTS at the centre from January 2001 to December 2005 was compiled and features such as: age, gender, drugs used, and outcome of treatment (defaulted, cured, died, or developed multidrug resistant-TB) were considered. Also the different personnel and infrastructure at the centre for the programme were also assessed. Results were analysed using Epi Info 6 statistical software, and P values < 0.05 were considered significant. RESULTS: Two hundred and seventy four (274) cases of pulmonary TB were registered at the centre during the study period, consisting of 100 (36.5%) females and 174 (63.5%) males with a statistically significant gender difference (P<0.001). The age range with the highest number of pulmonary tuberculosis cases was 31-40 years (24.8%; n=68), and the age range with the lowest number was 71 years and above (1.1%; n=3). Treatment outcome showed that 84.7% (n=232) completed treatment with cure; 2.5% (n=7) developed multidrug resistance at completion of treatment; 5.5% (n=15) defaulted; 3.3% (n=9) died in the course of treatment, and treatment in 11 people was still ongoing. CONCLUSION: The outcome of DOTS in the present study was impressive, and the programme should be extended to other rural communities; however, more efforts should be made towards the tracing of defaulters. PMID- 18390137 TI - Pattern of contraceptives choice among the married women attending the family planning clinic of a tertiary health institution. AB - BACKGROUND: Rampart population growth is the most fundamental problem of our times. It affects adversely the advancement of nations and the wellbeing of all peoples. The fertility rate in Nigeria remains high at a national average of about 5.2 children per woman. When a woman effectively uses a modern method of contraceptive she is less likely to be exposed to the hazards of grand multiparity and also unlikely to resort to dangerous illegal abortion. METHOD: This is a retrospective study, a review of the records of the family planning clinic of Ladoke Akintola University of Technology Teaching Hospital, Osogbo was undertaken. These were new clients coming for the first time to the family planning clinic of the institution from January, 2001 to December 2006. RESULT: A total of 1355 married women were coming for the first time to the family planning clinic within the study period, and their age range between 18 and 51 years with a mean of 33.5+/-6.1 years. 170 (12.5%) were Para 1, Para 2-4 were 855 (63.1%), while Para 5 and above were 330 (24.4%). The clients source of information about family planning was: family planning clinic personnel 1039 (76.7%), media (print and electronic) 152 (11.2%), friends and relatives 127 (9.4%), and community health workers 37 (2.7%). Intrauterine contraceptive device was the most chosen method by the clients 1011 (74.6%), while condom was the least chosen 3 (0.2%); 264 (19.5%), and 77 (5.7%) clients chose injectables and pills, respectively. CONCLUSION: An increase in the level of knowledge on modern contraceptive methods through mass and interpersonal communications could be one of the key strategies to increase the utilization rate of modern contraceptive methods. PMID- 18390139 TI - Ruptured tubal pregnancy in Makurdi, north central Nigeria. AB - BACKGROUND: Ruptured tubal pregnancy remains a major cause of early foetal wastage and maternal mortality in Nigeria and other developing countries. This study was done to determine the incidence, predisposing factors, clinical presentation and management options of ruptured tubal pregnancy at the Federal Medical Centre Makurdi. METHODOLOGY: A retrospective descriptive study of 35 cases of ruptured tubal pregnancy managed at the Federal Medical Centre, Makurdi, between January 2004 and December 2006 was conducted. RESULTS: The incidence of ruptured tubal pregnancy was 0.87% of total births (1 in 114 deliveries) and accounted for 94.6% of all ectopic pregnancies. There was a rising trend in the incidence of ruptured tubal pregnancy from 0.65% in 2004 to 1.09% in 2006. Identified risk factors include previous pelvic infection and previous induced abortions. Abdominal pain (100%), vaginal bleeding (65.7%), dizziness (62.9%) and amenorrhoea (57.1%) were the commonest symptoms. The ampulla was the site of rupture in 27(77.1%) patients. All patients had laparotomy. Unilateral total salpingectomy was done in 31(88.5%) patients. There was no maternal death in the cases managed. CONCLUSION: The incidence of ruptured tubal pregnancy is relatively low at the Federal Medical Centre Makurdi. Health education of women in the reproductive age on safe sex and eradication of unsafe abortion and early treatment of pelvic infections will prove useful preventive measures. PMID- 18390138 TI - Prevalence of risk factors for diabetes mellitus in a non-diabetic population in Jos, Nigeria. AB - BACKGROUND: Diabetes mellitus (DM) is assuming epidemic proportions worldwide, but probably more so in the developing world. Identification of risk factors for the development of type 2 diabetes mellitus is a necessary step in planning prevention programmes for diabetes mellitus. The objective of this study was to determine the frequency of risk factors for type 2 DM among inhabitants of Jos, a northern city on the Nigerian Plateau. METHOD: A district in central Jos was randomly picked. Census of the district was carried out to record the names of all eligible residents from 250 households selected systematically. A questionnaire was administered by trained interviewers. Socio-demographic data, family history of diabetes, and data on work related physical activity were recorded. Height, weight and waist and hip circumferences were also measured. BMI (kg/m2) and waist-hip ration were calculated. RESULTS: Of 902 subjects (= 15years of age), 825 (91.5%) responded. The mean (SD) age of 400 males and 422 females were respectively 36.4 (15.2) and 39.9 (17.3) years. About 50% of the respondents were inactive. 435 (52.7%) were currently taking alcohol. Twenty nine (3.5%) of the subjects admitted to parental history of DM. 177 (21.4%) were either overweight or obese. 32% of males and 86% of females had abnormal waist circumferences (WCE). 96 (23.8%) males and 316(74.9%) females had abnormal WHR. BMI correlated strongly and significantly with WHR (r = 0.64, p < 0.001) and WCE (r = 0.72, p < 0.001). CONCLUSION: Inactivity alcohol usage and excess weight appear to be dominant risk factors for development of type 2 DM in this group of upland Nigerians. PMID- 18390140 TI - Socio demographic profile and presentations of patients with ruptured gravid uterus in Calabar Nigeria. AB - BACKGROUND: To study the Socio-demographic profiles and mode of clinical presentations of with uterine rupture in pregnancy in Calabar, Nigeria. METHOD: Medical records of 67 patients managed for ruptured gravid uterus over 10 years in Maternity section of the University of Calabar Teaching Hospital Calabar, Nigeria were reviewed. RESULTS: An incidence of 1 in 213 of all deliveries during the period was established. Majority (49.2%) were aged between 31 to 40 years and 43.3% had no formal education. Unemployed patients were 29.9%, 42.3% did not book for antenatal care while 32.8% were attended to by traditional birth attendants (TBA) and in the churches. The Commonest clinical presentations were fetal heart rate abnormalities (52.2%) and maternal collapse (46.3%). Majority (50.7%) resulted from neglected obstructed labour. CONCLUSION: Ruptured uterus is a problem of ignorance among women of low socioeconomic group with most of them having unskilled and substandard care during pregnancy and delivery. Improvement in Socio-economic conditions and modification of some harmful cultural practices against women generally will reduce the problem of rupture uterus in our society. PMID- 18390141 TI - The clinical features of HIV/AIDS at presentation at the Jos University Teaching Hospital. AB - BACKGROUND: The clinical characteristics and consequences of HIV-I infection observed in studies from developed countries cannot be generalized across the multiple virus subtypes that circulate in sub-Saharan Africa in general and Nigeria in particular. This study was therefore conducted to determine the commonest symptoms and signs at presentation in HIV-infected individuals at the Jos University Teaching Hospital. METHOD: This descriptive study was conducted at the antiretroviral clinic of the Jos University Teaching Hospital, Jos, between May and October 2004. Two hundred (200) newly diagnosed and laboratory confirmed adult cases of HIV infection without prior antiretroviral drug use were recruited after obtaining informed consent. Each qualified patient had a comprehensive history taken with emphasis on the clinical symptoms and detailed physical examination performed by the researchers. The data collected were analyzed using a multipurpose computer programme, Epi-info 2000 version 1.1.3 (Atlanta GA, USA). RESULTS: There were 86 (43.0%) males and 114 (57.0%) females whose mean ages were 39.0 +/- 7.8 and 32.0 +/- 8.1 years, respectively. The major symptoms in the study population were: weight loss (65.5%), fever (41.5%), chronic cough (38.5%), diarrhea (32.0%), pruritus (13.0%) and body rash (12.5%). The major signs were pallor (25.0%), oral thrush (20.5%), wasting (20.0%), lymphadenopathy (18.0%), dermatitis (16.0%), hyperpigmented nails (13.5%) and finger clubbing (8.5%). CONCLUSION: The symptoms and signs of HIV/AIDS obtained were similar to those obtained by other workers from different parts of the world; however, the findings of hyperpigmented nails and finger clubbing have not been frequently reported for other populations. PMID- 18390142 TI - The prevalence and correlates of hypertension in a Theological College in Africa. AB - BACKGROUND: The Theological College is a peculiar setting. This is due to the nature of the job, studies, attitudinal leaning (faith) and influence. There has been no known work to date done on the cardiovascular status in institutions of this nature-both in the country and in the continent of Africa. This formed the basis for evaluating the prevalence of hypertension and its correlates in one of the foremost theological colleges in Africa, the Trinity Theological College, Umuahia. METHOD: The entire constituent working-student population in the sandwich program of August 2004 at the Trinity Theological College, Umuahia, were recruited in the study. Their biodata, demographic distribution, anthropometry, pulse rate and blood pressure measurements were recorded under standard conditions. Their lifestyle habits were evaluated. Structured questionnaires were used with self administered screening done by medical doctors. RESULTS: Eighty five subjects, with mean age 43.7 (+/- 9.9) years, were screened. The prevalence of hypertension was found to be 28.3%. Only 8 subjects admitted being known hypertensives, out of whom 6 were on treatment. Fifty three percent (45) of them checked their blood pressure irregularly, while 28 (33%) never did. Twenty six respondents (31%) admitted taking extra table salt, while 39 (46%) never engaged in any form of exercise. CONCLUSION: The prevalence of hypertension in this community is higher than that of the general Nigerian population. There is a great need for adequate health education especially with regard to regularity of blood pressure check and lifestyle modification, in the Theological Colleges, given the influence they wield in the larger society. PMID- 18390143 TI - Tinnitus and the prevalence of co-morbid psychological stress. AB - BACKGROUND: Tinnitus is a symptom of unknown pathophysiology with few therapeutic measures and may present with co-morbid psychological stresses necessitating psychiatric treatment. This study aims at determining the prevalence of depression and anxiety in tinnitus sufferers in our environment. METHOD: This is a one year (April 2006 March 2007) prospective study of out-patients presenting with tinnitus to our Ear, Nose and Throat clinic who were administered the Hospital Anxiety and Depression Scale (HADS) questionnaire until the sample size was reached. RESULTS: Questionnaires were administered to one hundred and four patients with tinnitus, 92 patients filled theirs correctly and these were analyzed. There were 42 (45.7%) males and 50 (54.3%) females (Table 1) with an age range of 20 to 78 years. Six hundred and eighty seven patients presented with various otologic ailments in the study period, of which 104 (15.1%) patients had tinnitus. The overall prevalence of depression was 17.4%, higher in females (9.8%) than males (7.6%). The overall prevalence of anxiety was 22.8% with males having a higher prevalence (11.9%) than females (10.9%). Three (3.2) patients had both depression and anxiety Eighty three (90.2%) patients were in the active and productive age group with 13 patients (prevalence of 14.1%) having depression and 20 patients (prevalence of 21.7%) with anxiety. CONCLUSION: We recommend the screening or assessment for psychological distress in tinnitus sufferers so that patients can be adequately treated. PMID- 18390144 TI - Utilization of primary health care facilities: lessons from a rural community in southwest Nigeria. AB - BACKGROUND: This study assessed service/organisational factors and clients' perceptions that influenced utilisation of Primary Health Care (PHC) facilities in a rural community in Nigeria. METHOD: A cross-sectional household survey in the community as well as key-informant interviews of opinion leaders and health care providers and participant observations of health facilities and utilisation pattern was used to collect data. RESULTS: Forty-four percent of respondents to the survey who were ill in the preceding six months visited a PHC facility for treatment, while others relied on self-medication/self-treatment. Education was positively associated with utilisation of PHC services (P<0.05). Maternal and child health (45.4%), prompt attention (23.0%), and appropriate outpatient (20.5%) services attracted respondents to use PHC services. Poor education about when to seek care, poverty, perceived high cost of PHC services, lack of drugs and basic laboratory services, and a regular physician on site at the facility were identified as barriers to utilisation. CONCLUSION: We conclude that community perceptions of poor quality and inadequacy of available services was responsible for low use of PHC services. PMID- 18390145 TI - Indications for surgical removal of the eye in adults: a five-year review. AB - BACKGROUND: To determine the indications and procedures used for removing the eye in adults in our environment. METHOD: A retrospective study of destructive ocular surgeries carried out between January 1999 and December 2003. Theatre and case records of adult patients that underwent surgical removal of the eye over a five year period were reviewed. Clinical and histological diagnoses were obtained as well as type of surgery and personal data. Results were analysed using simple percentages. RESULTS: The most common indication for removing the eye in adults was due to trauma in 47 (45.6%) cases while evisceration was the most common surgery performed in 91 (88.3%) cases. Males were three times more likely to lose an eye than females. In 74.8% of cases, the loss of an eye was in young persons less than 50 years. CONCLUSION: The economically productive age group are more likely to lose an eye. The reason for the loss of an eye in most cases is largely avoidable. Lack of education on ways to prevent ocular injuries and improved access to eye care services requires urgent attention. PMID- 18390146 TI - Oesophageal variceal band ligation using a saeed six-shooter multiband ligator; experience at Jos University Teaching Hospital, Nigeria: case report. AB - BACKGROUND: Oesophageal varices are common sequelae of cirrhosis, which when they bleed can be difficult to control. Oesophageal variceal band ligation, being the best modality for controlling variceal bleeding has not been common practice in West Africa, a region recognized to have a high prevalence of liver diseases. We present a case of band ligation of the varices of a patient that presented to our hospital with variceal haemorrhage, secondary to liver cirrhosis. METHOD: We started by presenting a summary of the presentation of a 31 year old man at the Jos University Teaching Hospital and how we established the diagnosis of grade IV bleeding oesophageal varices. The subject had four bands applied to the oesophageal varices using a saeed six shooter oesophageal Multi-band ligator (North Carolina, USA) using a forward-viewing GIF P30 gastroscope (Tokyo, Japan) with the bands mounted on an Opti-vu barrel-shaped piece. He had a total of three sessions after which he was maintained on propranolol. RESULT: The last recheck endoscopy demonstrated obliterated varices after which he was maintained on propranolol. CONCLUSION: We present a case of successful variceal band ligation of a cirrhotic with extensive oesophageal varices presenting in a resource constraint medical setting. PMID- 18390147 TI - Systemic onset juvenile chronic arthritis (JCA) in a Nigerian boy--a case report. AB - BACKGROUND: Juvenile chronic arthritis (JCA) is a chronic arthritis affecting children below age of 16 years. The systemic onset subgroup is also known as Still's disease. There are several distinct subgroups. There is paucity of literature of this disease entity in our environment due to under diagnosis of the disease. METHOD: The case note of this patient was retrieved. He was managed for 2 years in the adult rheumatology clinic after being transferred from the Paediatric unit. Relevant literature was reviewed. RESULT: There was a good response to immunosuppressive agents and low dose prednisolone. He was back at School and able to play with his mates after long withdrawal from School. CONCLUSION: Prompt referral of such cases to specialist centers will go a long way in determining the outcome of such patients. Prognosis is better with early presentation and appropriate management. PMID- 18390148 TI - Ileosigmoid knotting--a case for primary anastomosis. AB - BACKGROUND: Resection of gangrenous/non-viable bowel followed by primary anastomoses is known to be a viable option for treatment of ileosigmoid knotting. We here report a case of ileosigmoid knot and examined the options for treatment. METHOD: A case report of a patient with ileosigmoid knotting and discussion of relevant literature for surgical treatment. RESULT: A 26 year old man presented to the accident and emergency unit with two weeks' history of fever and alternating diarrhea and constipation followed by features of intestinal obstruction and later, generalized peritonitis. Plain abdominal X-rays showed dilated loops of bowel and multiple air-fluid levels. The diagnosis of ileosigmoid knotting was missed preoperatively. At exploratory laparotomy, ileosigmoid knot was encountered with non-viable segments of the sigmoid colon and ileum. Resection of the non-viable bowel was carried out with primary anastomoses with good postoperative outcome. CONCLUSION: Primary anastomoses after resection of gangrenous bowel is a safe surgical treatment option in ileosigmoid knotting. PMID- 18390149 TI - Clinical photographic quiz in clinical microbiology. PMID- 18390150 TI - [Haemolytic uremic syndrome: why Argentina?]. PMID- 18390151 TI - [Multiplication of Brucella abortus and production of nitric oxide in two macrophage cell lines of different origin]. AB - Brucella abortus is a bacterium which causes abortions and infertility in cattle and undulant fever in humans. It multiplies intracellularly, evading the mechanisms of cellular death. Nitric oxide (NO) is important in the regulation of the immune response. In the present work, we studied the ability of three B. abortus strains to survive intracellularly in two macrophage cell lines. The bacterial multiplication in both cell lines was determined at two different times in UFC/ ml units. Moreover the inoculated cells were also observed under light field and fluorescence microscopy stained with Giemsa and acridine orange, respectively. The stain of both cellular lines showed similar results with respect to the UFC/ml determination. The presence of B. abortus was confirmed by electronic microscopy. In both macrophage cell lines inoculated with the rough strain RB51, the multiplication diminished and the level of NO was higher, compared with cells inoculated with smooth strains (S19 and 2308). These results suggest that the absence of O-chain of LPS probably affects the intracellular growth of B. abortus. PMID- 18390152 TI - Whole-cell protein profiles are useful for distinguishing enterococcal species recovered from clinical specimens. AB - Whole-cell protein analysis was performed for differentiating 150 enterococcal isolates to the species level, which had previously been identified by extended phenotypic conventional tests. Whole-cell protein profile (WCPP) showed a high degree of similarity within species and comparison between species revealed important differences in band profiles. All Enterococcus faecalis and Enterococcus faecium isolates were properly located into their corresponding species, regardless of their clinical source and susceptibility pattern. Moreover, WCPP allowed relocation of some isolates that had erroneously been identified by the usual conventional scheme (i.e. two atypical arginine-negative E. faecalis isolates). WCPP proved to be a simple method to ascertain the various enterococcal species, especially those other than E. faecalis, and may be a suitable tool for high-complexity or reference clinical laboratories. PMID- 18390153 TI - Biofilm formation by Stenotrophomonas maltophilia isolates from device-associated nosocomial infections. AB - Medical devices are often colonized by bacteria which may cause severe infections. The aim of this work was to evaluate biofilm formation by S. maltophilia isolates from device-associated nosocomial infections. The 13 local isolates exhibited different capacities of biofilm formation on hydrophilic and hydrophobic surfaces. All isolates formed strong biofilms in polystyrene microplates, while strong, moderate or weak biofilms were detected in borosilicate (BS) or polypropylene (PP) tubes. The proficiency of biofilm formation was better evaluated by the level of crystal violet staining expressed relative to the final culture density. The microscopic analysis of biofilms formed on glass coverslips revealed the presence of a matrix of exopolysaccharides and microcolonies typical of biofilm architecture. Isolates with increased adhesion to BS showed larger microcolonies. According to our results, twitching correlated well with attachment to the three abiotic surfaces tested, while swimming only showed a slight correlation with biofilm formation on PP. Poor correlation was observed between cell surface hydrophobicity and biofilm formation. One of the highest biofilm-producing isolates adhered to urethral catheters of different materials, and exhibited an increased resistance to oxidative stress, one of the common stresses encountered by bacteria during the infection process due to the immune response. PMID- 18390154 TI - Fungal peritonitis in patients on peritoneal dialysis: twenty five years of experience in a teaching hospital in Argentina. AB - Fungal peritonitis is a rare but serious complication of peritoneal dialysis. The aim of this study was to analyze peritonitis rates, associated factors, clinical course, microbiological aspects, therapeutic regimens, and outcome of patients with fungal peritonitis in the dialysis center of a teaching hospital over the last 25 years. A hundred and eighty three episodes of peritonitis were detected and microbiologically documented in 57 patients. Fungi were identified in eight episodes (4.37%) occurring in seven female patients. The fungal peritonitis rate was 0.06 episodes/patient-year. Gram and Giemsa stains were positive in five out of eight dialysate fluids. The causative microorganisms were: Candida albicans in five episodes, and Candida parapsilosis, Candida glabrata, and Neosartorya hiratsukae in the remaining three. Antibiotics were administered to all but one patient, within 3 months before fungal peritonitis was detected. All patients required hospitalization, and antifungal therapy was administered in all episodes. The Tenckhoff catheter was removed in seven out of eight fungal peritonitis. All patients recovered from the fungal episodes. In the group of patients studied, it is concluded that recent exposure to antibiotics and female sex, were strongly associated with the development of fungal peritonitis by yeasts. The peritonitis caused by the environmental filamentous fungus did not require antibiotic pressure. Direct microscopy of the dialysate pellet was extremely useful for the prompt management of the fungal episode. Fungal peritonitis preceded by multiple episodes of bacterial peritonitis always determined the definitive dropout of the patient from the peritoneal dialysis program. Patients with de novo yeast-related peritonitis could continue on the program. PMID- 18390155 TI - [Tinea nigra palmaris: a clinical case in Argentina]. AB - A clinical case of a female patient with a black spot on the palm of her left hand is presented. The infection was due to a black fungus identified as Hortaea werneckii, the aetiological agent of tinea nigra palmaris. This infection can be easily diagnosed and it is important to establish the differential diagnosis from other skin pathologies. Normally, the treatment has a successful outcome. In this case, the patient was treated with econazole locally applied during one month, with complete remission of the lesions. In conclusion, the early diagnosis of this disease is very important since the mycology procedures are fast and non invasive and cure is obtained with local treatment. PMID- 18390156 TI - Seasonal fluctuations in prevalence of dog intestinal parasites in public squares of Mar del Plata city, Argentina and its risk for humans. AB - The aim of this work is to analyze whether there is a seasonal prevalence of parasites in dog feces from public squares in Mar del Plata city, Argentina, and to evaluate the climatic conditions that promote the development of parasites and allow the transmission to people. The study was performed in 21 squares from June 2001 to May 2002. Samples were processed by the Willis technique. Differences in prevalence of parasites were examined for significance by the Chi-square test. Climatic data were obtained from the Library of the National Meteorological Service of Argentina. Total parasitic prevalence was higher in winter than in summer. The prevalence of Ancylostoma spp. was higher in the summer-autumn period. For Toxocara canis, the prevalence was higher in winter whereas for Trichuris vulpis, it was higher in winter, spring and summer. This work shows high prevalence of total parasites throughout the year. For Ancylostoma spp., summer and autumn might be the seasons with higher sanitary risk. On the other hand, T. canis could present the higher risk to people in winter and, T. vulpis, would be transmitted throughout the whole year. The seasonal variation in prevalence of dog parasites results in continuous exposure to people visiting the squares, not only Mar del Plata residents but also tourists from other regions of Argentina and the world, with at least one species of parasite with sanitary risk. PMID- 18390157 TI - [Insecticide proteic crystals from a native isolate of Bacillus thuringiensis]. PMID- 18390158 TI - [Leprosy and paracoccidioidomycosis]. PMID- 18390159 TI - [Antimicrobial resistance of Escherichia coli isolated from pigs in Argentina]. AB - Antimicrobial resistance of Escherichia coil isolated from pigs in Argentina. Sixty-nine Escherichia coli isolates from healthy pigs or with clinical signs non compatible with diarrhea caused by this microorganism, were studied. The purpose was to determine the resistance profile against antimicrobials frequently used in veterinary and human medicine. The agar diffusion method was used. High resistance percentages against antimicrobials used in swine farms such as ampicillin, streptomycin and tetracycline were observed, as well as against trimetoprim-sulfametoxazole and chloramphenicol, compounds that were stopped being used several years ago. Sixty two percent of isolates showed multidrug resistance. The results obtained in this work corroborate the hypothesis that the phenotypic distribution of resistance and possibly that of its genetic determinants, are directly influenced by the antimicrobial treatments used. PMID- 18390160 TI - [Rapid identification and susceptibility to killer toxins of yeasts isolated from non-systemic mycoses]. AB - Rapid identification and susceptibility to killer toxins of yeasts isolated from non-systemic mycoses. The use of quick and reliable yeast identification methods, as well as the development of new antifungal agents with more specific targets, will enable a more efficient treatment of mycoses. In the present work, a total of 53 clinical isolates obtained from non-systemic infections in Neuquen Hospitals and an ophthalmologic clinic in Buenos Aires during 2005, were identified by means of a rapid molecular method (ITS1-5.8S ADNr-ITS2 PCR-RFLP). Additionally, the killer susceptibility of the isolates was tested against reference and indigenous killer yeasts on plate tests. Eight yeast species were identified among the clinical isolates: Candida albicans (52%), Candida parapsilosis (17%), Candida tropicalis (10%), Candida krusei (5%), Candida glabrata (4%), Candida guilliermondii (4%), Kluyveromyces lactis (4%) and Saccharomyces cerevisiae (4%). Sixty-nine percent of the isolates corresponding to the predominant species (C. albicans) were related to vaginal infections. On the other hand, 61% of the yeasts associated with ocular infections were identified as C. parapsilosis. Two indigenous killer isolates DVMais5 and HCMeiss5, belonging to Pichia anomala and P. kluyveri respectively, exhibited the broadest killer spectrum against clinical isolates. PMID- 18390161 TI - Simplified prediction of Staphylococcus aureus growth in a cooked meat product exposed to changing environmental temperatures in warm climates. AB - In this work, a simplified method is used to estimate the growth of Staphylococcus aureus in a pasteurized meat product left for several hours at environmental temperatures (diurnal time) in warm climates of different cities in Argentina. Hourly temperature data for a warm January (the hottest month of the year) day, and literature data on the kinetics of S. aureus growth inoculated in a pasteurized meat product were used for calculations. As shown by results, if a cooked meat product is left exposed to environmental temperature at diurnal time, predictions made when using a constant temperature value (i.e. average daily) may not be accurate. Growth estimations in contaminated food left under ambient conditions during diurnal time, should consider the changing environmental temperature for correct results. PMID- 18390162 TI - [Bacterial diversity in a fluidized bed bioreactor (FBR) treating gasoline contaminated groundwater]. AB - Bacterial diversity in a fluidized bed bioreactor (FBR) treating gasoline contaminated groundwater. The main objective of this research project was to determine the bacterial diversity during the process of bioremediation of water contaminated with gasoline in a fluidized bed reactor at Mayaguez, PR. Isolation and characterization of bacterial populations from the bioremediation system was performed on R2A medium. Morphological tests included cellular and colonial shape and reaction to Gram coloration. Physiological properties were determined by using carbon utilization profiles (Biolog) and by the ability of axenic cultures to use gasoline as the sole carbon source. Molecular characterization was performed by BOX-PCR and 16S rDNA sequence analysis (ARDRA). From a total of 162 distinctive isolates, 75% were gram-negative bacilli, 19% gram-positive bacilli, 5% gram-negative cocci and 1% gram-positive cocci. The 162 axenic cultures corresponded to 90 different genetic groups; 53% of which included strains with growth in gasoline as sole carbon source. The 86 strains capable of growing in gasoline corresponded to 52 different amplification patterns in BOX-PCR; which were not metabolically related (Biolog system). The high degree of microbial diversity in the FBR allowed efficient and stable hydrocarbon removal throughout the operation of the system. PMID- 18390163 TI - The role of organizational factors in the adoption of healthcare information technology in Florida hospitals. AB - This study examines whether specific organizational characteristics, such as hospital size, geographic location (urban versus rural), system membership (stand alone versus system-affiliated), and tax status (for-profit versus non-profit), influence adoption of healthcare information technologies (HIT) in hospitals. We hypothesize the above organizational characteristics to be related to hospitals' adoption of clinical, administrative, and strategic HIT, as well as all HIT in general. Using survey data collected from 98 Florida hospitals, we demonstrate that hospital size, system membership, and tax status, but not geographic location, are systematically related to HIT adoption, and that such factors explain about 28-41% of the adoption variance. A mixed pattern of effects emerge for clinical, administrative, and strategic HIT. For instance, hospital size appears to be less relevant for administrative HIT, where its effect is compensated by those of system membership and tax status. Implications for future HIT research and practice are discussed. PMID- 18390164 TI - Using queuing theory to analyse the government's 4-H completion time target in accident and emergency departments. AB - This paper uses a queuing model to evaluate completion times in Accident and Emergency (A&E) departments in the light of the Government target of completing and discharging 98% of patients inside 4 h. It illustrates how flows though an A&E can be accurately represented as a queuing process, how outputs can be used to visualise and interpret the 4-h Government target in a simple way and how the model can be used to assess the practical achievability of A&E targets in the future. The paper finds that A&E targets have resulted in significant improvements in completion times and thus deal with a major source of complaint by users of the National Health Service in the U.K. It suggests that whilst some of this improvement is attributable to better management, some is also due to the way some patients in A&E are designated and therefore counted through the system. It finds for example that the current target would not have been possible without some form of patient re-designation or re-labelling taking place. Further it finds that the current target is so demanding that the integrity of reported performance is open to question. Related incentives and demand management issues resulting from the target are also briefly discussed. PMID- 18390165 TI - Optimizing sterilization logistics in hospitals. AB - This paper deals with the optimization of the flow of sterile instruments in hospitals which takes place between the sterilization department and the operating theatre. This topic is especially of interest in view of the current attempts of hospitals to cut cost by outsourcing sterilization tasks. Oftentimes, outsourcing implies placing the sterilization unit at a larger distance, hence introducing a longer logistic loop, which may result in lower instrument availability, and higher cost. This paper discusses the optimization problems that have to be solved when redesigning processes so as to improve material availability and reduce cost. We consider changing the logistic management principles, use of visibility information, and optimizing the composition of the nets of sterile materials. PMID- 18390166 TI - Modelling of errors in databases. AB - A lot of time and energy are expended assembling national databases containing information about health care processes and outcomes. Unfortunately, given the complexity of the data gathering procedures involved, errors occur. This inevitably leads to problems when it comes to the analysis of data from such sources. Indeed, sometimes it is very much a matter of faith that summary statistics represent a true reflection of the facts. On the assumption that one knows the rates at which different forms of errors occur, mathematical modelling methods can be used to obtain estimates of the effects of such errors on the estimates that would be derived for summary statistics associated with an erroneous data base. PMID- 18390167 TI - Stratified random sampling for estimating billing accuracy in health care systems. AB - This paper presents a stratified random sampling plan for estimating accuracy of bill processing performance for the health care bills submitted to third party payers in health care systems. Bill processing accuracy is estimated with two measures: percent accuracy and total dollar accuracy. Difficulties in constructing a sampling plan arise when the population strata structure is unknown, and when the two measures require different sampling schemes. To efficiently utilize sample resource, the sampling plan is designed to effectively estimate both measures from the same sample. The sampling plan features a simple but efficient strata construction method, called rectangular method, and two accuracy estimation methods, one for each measure. The sampling plan is tested on actual populations from an insurance company. Accuracy estimates obtained are then used to compare the rectangular method to other potential clustering methods for strata construction, and compare the accuracy estimation methods to other eligible methods. Computational study results show effectiveness of the proposed sampling plan. PMID- 18390168 TI - Understanding and reducing the medication delivery waste via systems mapping and analysis. AB - The enormous gaps in the knowledge required to understand medication errors and their related costs (or wastes) in all hospital settings have become a growing national concern. Such gaps are often the major reasons causing risk for patient safety and creating waste to the hospital. However, medication delivery system cannot be successfully improved and implemented without a clear understanding of various process flows running around the entire hospital system. This paper presents a systems mapping and analysis method to help understand and reduce the medication delivery waste. The effectiveness of our method is illustrated by a case study that we conducted for the medication delivery process at Bozeman Deaconess Hospital, MT. PMID- 18390169 TI - Is more better? An analysis of hospital outcomes and efficiency with a DEA model of output congestion. AB - This paper applies a new methodology to the study of hospital efficiency and quality of care. Using a data set of hospitals from several states, we jointly evaluate desirable hospital patient care output (e.g., patient stays) and the simultaneous undesirable output (e.g., risk-adjusted patient mortality) that occurs. With a DEA based approach under two different sets of assumptions, we are able to include multiple quality indicators as outputs. The results show that lower technical efficiency is associated with poorer risk-adjusted quality outcomes in the study hospitals. They are consistent with other studies linking poor quality outcomes to higher cost. PMID- 18390170 TI - Using the Statecharts paradigm for simulation of patient flow in surgical care. AB - Computer simulation of patient flow has been used extensively to assess the impacts of changes in the management of surgical care. However, little research is available on the utility of existing modeling techniques. The purpose of this paper is to examine the capacity of Statecharts, a system of graphical specification, for constructing a discrete-event simulation model of the perioperative process. The Statecharts specification paradigm was originally developed for representing reactive systems by extending the formalism of finite state machines through notions of hierarchy, parallelism, and event broadcasting. Hierarchy permits subordination between states so that one state may contain other states. Parallelism permits more than one state to be active at any given time. Broadcasting of events allows one state to detect changes in another state. In the context of the peri-operative process, hierarchy provides the means to describe steps within activities and to cluster related activities, parallelism provides the means to specify concurrent activities, and event broadcasting provides the means to trigger a series of actions in one activity according to transitions that occur in another activity. Combined with hierarchy and parallelism, event broadcasting offers a convenient way to describe the interaction of concurrent activities. We applied the Statecharts formalism to describe the progress of individual patients through surgical care as a series of asynchronous updates in patient records generated in reaction to events produced by parallel finite-state machines representing concurrent clinical and managerial activities. We conclude that Statecharts capture successfully the behavioral aspects of surgical care delivery by specifying permissible chronology of events, conditions, and actions. PMID- 18390171 TI - [An experimental study of the culture, isolation and biological characteristics of rat adipose tissue-derived stromal cells in vitro]. AB - OBJECTIVE: To establish a method for the isolation and culture of rat adipose tissue-derived stromal cells (ADSCs) and explore some biological characteristics of the acquired ADSCs. METHODS: Adipose tissues were isolated from the inguinal fat of SD rats. Primary ADSCs were obtained by the method of collagenase I digestion, inoculated, cultured in the Dulbecco modified Eagle medium with 10% fetal bovine serum, and subcultured at the right moment. The morphology and proliferation characteristics of the cells were observed under the inverted phase contrast microscope every day. Their growth curves were detected and experiments of freezing and resuscitation performed. The third passage ADSCs were induced into osteoblasts by osteogenic inducing fluid and into adipocytes by adipogenic inducing fluid. The osteogenic phenotypes were examined by Von Kossa staining and the adipocytes by Oil Red O staining. RESULTS: ADSCs were successfully obtained and cultured from the rat adipose tissue. They appeared fibroblast-like and could proliferate rapidly in vitro, the third passage having the most active proliferative ability. Calcium nodes characteristic of osteoblasts were observed in the ADSCs on Von Kossa staining after induction with dexamethasone, ascorbic acid and beta-sodium glycerophosphate, and red-stained fats characteristic of adipocytes were noted in the cytoplasm on Oil Red O staining after induction with IBMX, indomethacin and insulin. The ADSCs showed no significant decrease in their proliferation activity and capability of differentiating into diverse cell types after cryopreserved in liquid nitrogen for a month. CONCLUSION: A simple and effective method for the isolation and culture of rat ADSCs was successfully established. The ADSCs obtained could grow and proliferate rapidly in vitro, capable of differentiating into diverse cell types, easy to be preserved and promising to be seed cells for cell therapy and tissue engineering. The procedure of schizolysising erythrocytes with NH4Cl could be omitted in the isolation of the rat ADSCs and dexamethasone is not indispensable in the induction of ADSCs into adipocytes. PMID- 18390172 TI - [Development and clinical application of male human papillomavirus genotyping by membrane DNA chip]. AB - OBJECTIVE: To develop a new method for the detection of male human papillomavirus (HPV) genotypes and to investigate its clinical application value. METHODS: With computer assistance and based on the classical common primers MY09/11, modified PGMY09/11 with 23 HPV subtypes for PCR and Genbank data on HPV, we designed probes for the simultaneous detection of 18 high-risk subtypes (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 83 and MM4) and 5 low-risk subtypes (HPV-6, 11, 42, 43 and 44) and fixed them to the special membrane to make a DNA chip. A total of 112 male urethral samples were collected with swabs and studied for the clinical value. Meanwhile the single subtypes of HPV positive were sequenced and the standard samples detected for their sensitivity. RESULTS: Of the total number, 25 samples were found to be HPV positive, 13 single HPV infection and 12 multiple infection. Nine HPV gene subtypes were noted in the samples: 6, 11, 16, 18, 33, 35, 43, 56 and 73, with sensitivity up to 10 copies of HPV DNA. CONCLUSION: Human papillomavirus genotyping by the membrane DNA chip is applicable to the diagnosis of male HPV infection as well as to the related epidemic and etiological investigation. PMID- 18390173 TI - [Advanced glycation end products and their receptors elevate the activity of endothelin-1 in rat cavernosum]. AB - OBJECTIVE: To investigate the role of advanced glycation end products (AGEs) and their receptors (RAGE) in the pathogenesis of diabetic mellitus erectile dysfunction (DMED) and the effects of AGEs and RAGE on the activity of endothelin 1 (ET-1) in rat cavernosum. METHODS: Forty male Sprague-Dawley rats were taken at random to construct 2 groups of diabetes mellitus (DM) models of equal number, one given free access to water and the other administered aminoguanidine hydrochloride (DM + AG) in water at the dose of 1 g/L. Another 20 male SD rats were equally divided into a normal control and an AG control group. After 8 weeks, the cavernosum tissues were harvested from all groups of rats, part of the isolated penile tissues homogenated to detect the content of AGE-peptide (AGE-P) and the activity of ET-1, and the AGEs and RAGE in the rest of the penile tissues analyzed by immunohisto- chemical assay. RESULTS: Compared with the normal controls, the expressions of AGEs and RAGE, the content of AGE-P and the activity of ET-1 in the cavernosum tissues were significantly high in the DM group (P < 0.05), while the administration of AG to the DM rats reversed the above results. No significant difference was observed between the normal control and AG control groups in any of the data (P > 0.05). CONCLUSION: In DM conditions, the joint effect of AGEs and RAGE may elevate the activity of ET-1 in rat cavernosum and thus promote the development of DMED. PMID- 18390174 TI - [Curcumin inhibits the expression of vascular endothelial growth factor and androgen-independent prostate cancer cell line PC-3 in vitro]. AB - OBJECTIVE: To study the effects of curcumin on the expression of the vascular endothelial growth factor (VEGF) and androgen-independent prostate cancer cell line PC-3, and to explore its anticarcinogenic mechanism. METHODS: PC-3 cells were treated with curcumin at the concentration of 0, 6.25, 12.5, 25 and 50 micromol/L respectively. Then the cell activity was assayed by dyed rate of Typan blue and MTT at 12, 24, 36, 48, 72 and 96 hours, the cell cycle and morphological changes observed by flow cytometry (FCM) and electronic microscopy at 24 hours, the VEGF mRNA expression measured by semi-quantitative RT-PCR, and the secreting protein levels of VEGF in the supernatants determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The growth of PC-3 cells was suppressed obviously by curcumin in a dose- and time-dependent manner in vitro. There were significant differences in inhibition rate among different concentration and time groups (P < 0.01). Furthermore, curcumin arrested the cell cycle of PC-3 cells in the G2/M phase in a dose-dependent manner (P < 0.01). The percentages of apoptotic cells were significantly higher in different concentration groups than in the controls (P < 0.01). Apoptosis-associated morphological changes were observed in PC-3 cells at 24 hours, and a marked decline in the expression of VEGF was noted after the exposure to different concentrations of curcumin within 24 hours. CONCLUSION: Curcumin can suppress the growth of PC-3 cells, promote their apoptosis and arrest their cell cycle in the G2/M phase, and reduce the expression of VEGF mRNA and proteins, which may sever to explain its inhibitory effect on tumor and angiogenesis. PMID- 18390175 TI - [All-trans retinoic acid enhances bystander effect of suicide-gene therapy against androgen-unresponsive prostate cancer]. AB - OBJECTIVE: To investigate the enhancing effect of all-trans retinoic acid (ATRA) on the bystander effect of the herpes simplex virus thymidine kinase(HSV TK)/ganciclovir (GCV) against androgen unresponsive prostate cancer. METHODS: The bystander effect of the HSV-TK/GCV system was measured by methyl thiazolyl tetrazolium (MTT) assay on PC-3 cells before and after ATRA treatment. The growth and the histopathology of transplant tumors were observed in 4 groups of nude mice with prostate cancer. RESULTS: ATRA augmented significantly the bystander effect of the HSV-TK/GCV system by reducing TK positive PC-3 cells from 50% to 30% (P < 0.05). HSV-TK showed an inhibiting effect, while ATRA with the HSV TK/GCV system produced significant effect on prostate cancer 1 week earlier than the former (P < 0.05). CONCLUSION: ATRA can argument the in vivo and in vitro bystander effect of the HSV-TK/GCV system in the treatment of androgen unresponsive prostate cancer. PMID- 18390176 TI - [Effect of (CAG) n polymorphism of androgen receptor gene on hormonal male contraception]. AB - OBJECTIVE: To study (CAG) n polymorphism of the androgen receptor (AR) gene in responders and non-responders of male volunteers who received testosterone undecanoate intramuscular injection for contraception and to explore the effect of the polymorphism on hormonal male contraception. METHODS: Twenty-nine non responders and 34 responders were enrolled in this study as a test and a control group respectively. The numbers of CAG sequence repeats were determined by PCR and DNA sequencing, and the effect of (CAG) n polymorphism on hormonal male contraception was analyzed. RESULTS: The means of CAG repeats of the test and the control group were 23.62 and 22.97, with no significant difference in between (P > 0.05). The short CAG repeats (n < or = 22) constituted 51.7% in the test group and 50% in the control, while the long ones (n > 22) accounted for 48.3% and 50% , respectively. The short and the long group had a similar distribution. No association was found between CAG repeats and sperm concentration. With FSH > 0.2 IU/L, the probability of azoospermia in the long CAG repeat group was 1.5 times that of the short one. CONCLUSION: CAG repeats in the AR gene presented polymorphism in the subjects, with no significant difference between the responders and non-responders. Further investigation has yet to be performed into the relationship of hormonal male contraception with CAG repeats or other factors. PMID- 18390177 TI - [Diagnosis and treatment of non-specific granulomatous prostatitis: a report of 32 cases]. AB - OBJECTIVE: To discuss the diagnosis and treatment of non-specific granulomatous prostatitis (NSGP). METHODS: Thirty-two cases of NSGP were diagnosed by puncture biopsy under transrectal ultrasound (TRUS) and treated with antibiotics and other medicines from September, 2000 to May, 2006. RESULTS: Pathomorphologically, NSGP was basically characterized by granuloma with vessels or grand alveoli in the center. The mean follow-up was 24 months. Urination irritation and obstruction were improved. Q(max) was increased to 15.0-24.0 ml/s, and in 3 cases of urinary retention, to 12.0, 14.5 and 16.5 ml/s, respectively. Digital rectal examination (DRE) indicated a reduced size and softened texture of the prostate induration. PSA was decreased to 1.3-11.5 microg/L. Four cases experienced relapse but were cured after retreated. No prostate cancer was observed. CONCLUSION: NSGP can be definitely diagnosed by puncture biopsy under TRUS and effectively relieved by antibiotics with the alpha-receptor blocker. In case of serious obstruction complicated by urinary retention, transurethral electrotomy can be considered. PMID- 18390178 TI - [Value of transrectal ultrasonography in the diagnosis of midline prostatic cysts]. AB - OBJECTIVE: To assess the value of transrectal ultrasonography (TRUS) in the diagnosis of midline prostatic cysts. METHODS: We retrospectively analyzed the TRUS manifestations of 87 cases of midline prostatic cysts. RESULTS: Of the total number, 33 cases were diagnosed as Mullerian duct cysts, 21 cases ejaculatory duct cysts and the other 33 cases undifferentiated midline prostatic cysts; 19 cases had dilated seminal vesicles, 19 seminal vesicle agenesis, 9 seminal vesiculitis and 5 dilation of the ejaculatory duct. CONCLUSION: TRUS, convenient, sensitive, safe and non-invasive, is a desirable method for the diagnosis of midline prostatic cysts. PMID- 18390179 TI - [Effects of prepubertal exposure to diethylstilbestrol on testicular development and function of SD rats]. AB - OBJECTIVE: To investigate the effects of prepubertal exposure to diethylstilbestrol (DES) on the testicular development and function of Sprague Dawley (SD) rats. METHODS: Ninety 21-day-old male SD rats were randomly and equally divided into 4 experimental groups (Da, Db, Dc and Dd), which were injected with DES dissolved in corn oil at the dose of 0.01, 0.1, 1.0 and 10.0 microg/(kg x d) from postnatal day (PND) 22 to 35, and a control group (C), which received vehicle only. The testicular development of all the rats was observed, and their testes were harvested in the stages of late puberty (PND 50), sexual maturity (PND 64) and adulthood (PND 130) respectively to determine the weight and histological features of the testis and examine the quality of the sperm in the epididymal cauda of the PND 130 rats. RESULTS: The testis descent in the C, Da, Db, Dc and Dd groups occurred on PND 26.17 +/- 1.94, 26.83 +/- 1.47, 28.68 +/ 1.03, 33.50 +/- 1.87 and 41.50 +/- 2.74 respectively, significantly delayed in the Db, Dc and Dd groups compared with the C group (P < 0.05 or P < 0.01). On PND 50, the unilateral testis weights in the C, Da, Db, Dc and Dd groups were (1.38 +/- 0.01) g, (1.38 +/- 0.12) g, (1.30 +/- 0.14) g, (0.86 +/- 0.18) g and (0.73 +/ 0.27) g respectively, significantly less in the Dc and Dd groups than in the C group (P < 0.01). Compared with the C group, there was a slight decrease in the number of the cells in the epithelia of a few seminiferous tubules in the Db group on PND 50, maldevelopment of seminiferous tubules, reduced cell number in seminiferous epithelia, blocked spermatogenesis and aplasia of Leydig cells in the Dc and Dd groups in a dose-dependent manner. On PND 64, the unilateral testis weights in the C, Da, Db, Dc and Dd groups were (1.60 +/- 0. 06) g, (1.62 +/- 0.11) g, (1.58 +/- 0.08) g, (1.47 +/- 0.10) g and (0.99 +/- 0.37) g respectively, significantly less in the Dc and Dd groups than in the C group (P < 0.05 or P < 0.01), and the histological alteration of the testis in the Dc and Dd groups was similar to or less than that on PND 50. On PND 130, no statistic difference was observed either in unilateral testis weight or in the histological features of the testis between any experimental group and the control (P > 0.05). The sperm concentration in the epididymal cauda in the C, Da, Db, Dc and Dd groups were (73.00 +/- 16.90) x 10(6)/ml, (68.00 +/- 19.67) x 10(6)/ml, (68.67 +/- 12.15) x 10(6)/ml, (35.17 +/- 15.64) x 10(6)/ml and (19.13 +/- 5.17) x 10(6)/ml, significantly lower in the Dc and Dd groups than in the C group (P < 0.01). There was a significant decrease in sperm motility in the Dd group (P < 0.01), the percentage of grade a sperm in the Db, Dc and Dd groups (P < 0.05) and the percentage of grade b sperm in the Dd group (P < 0.01). CONCLUSION: Prepubertal exposure to low dose of DES (0.01 microg/[kg x d] x 14 d) does not significantly affect the testicular development and function of SD rats, while high dose (1.0 10.0 microg/[kg x d] x 14 d) has significant short- (PND 50 and 64) or long-term (PND 130) toxic effect, which increases with dose and decreases with age. The mechanism of the toxic effect involves the insults to the development and function of Leydig and Sertoli cells. PMID- 18390180 TI - [Clinical efficacy of combined L-carnitine and acetyl-L-carnitine on idiopathic asthenospermia]. AB - OBJECTIVE: To evaluate the clinical efficacy of combined L-carnitine and acetyl-L carnitine on idiopathic asthenospermia. METHODS: Thirty patients with idiopathic asthenospermia were treated by combined L-carnitine and acetyl-L-carnitine for 3 months after failure to respond to the integrated therapy of Chinese and Western medicine. Semen samples were obtained, analyzed and graded according to the WHO laboratory manual before and after the treatment. Twenty-five of the patients completed the whole study. RESULTS: Combined L-carnitine and acetyl-L-carnitine statistically improved the sperm vitality (24.89 +/- 12.28)%, grade a + b sperm motility (16.04 +/- 8.33)% and the total sperm count per ejaculate (76.79 +/- 43.14) x 10(6), with a significant difference from pre-treatment (P < 0.05). CONCLUSION: Combined L-carnitine and acetyl-L-carnitine has a supplementary effect in the treatment of idiopathic asthenospermia and improves the semen quality of the patient. PMID- 18390181 TI - [Urethroplasty with tunica vaginalis flap for treating hypospadias with hydrocele: a report of 9 cases]. AB - OBJECTIVE: To explore the treatment of hypospadias with hydrocele. METHODS: Nine patients with hypospadias complicated by hydrocele underwent urethroplasty with tunica vaginalis flap. RESULTS: Satisfactory results were achieved in 7 cases, all with a normal anatomic position of urethral meatus, perfect urination and no infection. Local infection and meatal stricture occurred in 1 case, which was restored to normal by urethral sounding. Scrotal urinary fistula developed in another. CONCLUSION: With minimum surgical trauma, conveniently available material and fast tissue repair, urethroplasty with tunica vaginalis flap is a desirable method for the treatment of hypospadias with hydrocele. PMID- 18390182 TI - [Relationship between RhoA/Rho-kinase signaling pathway and penile erection]. AB - Erectile dysfunction (ED) has been plaguing men for a long time and the incidence of this disease is as high as 52% among males aged between 40 and 70. Recent discovery has shown a connection between the RhoA/Rho-kinase signaling system and ED. This paper reviews the progress in the study of RhoA/Rho-kinase signaling pathway, expounds its mechanism in penile erection and provides a base for further research on the role of RhoA/Rho-kinase signaling pathway in penile erection. PMID- 18390183 TI - [Toxicity of acrylamide on male reproduction]. AB - Acrylamide is a common chemical material, extensively used in industry and scientific experiments. Recently, it has been reported that starchy food cooked at high temperature can produce acrylamide. Acrylamide monomer has several toxic effects and the extensive concern for its toxicity has arisen with the finding of acrylamide formation in some processed foods. Researches have shown that acrylamide monomer can cause reproductive toxicity, including toxic effects on male reproductive behavior, male reproductive endocrine function and spermatogenesis. The mechanisms may include the effects of acrylamide on Leydig cells, the formation of motor protein/ chromosomal/DNA alkylation and damage by oxidative stress. PMID- 18390184 TI - [A novel class of testis-specific small RNAs: piRNAs]. AB - Eukaryotes produce various types of 19-30 nt small RNAs, which act as guides to the regulation of gene expressions, such as mRNA degradation and translational repression. The Argonaute family members related to small RNA functions fall into 2 subfamilies. One is the AGO subfamily, whose 4 members distribute widely, confirmedly bind to miRNAs and siRNAs and inhibit the expression of target mRNAs through a pathway like RNA interference. The other is the PIWI subfamily, including PIWI, Aubgine (AUB) and AGO3, exclusively expressed in the testis. Recently, four research groups have isolated a new class of small RNAs from the mammalian testis, which interacts with the PIWI subfamily, hence named piwi interfering RNAs (piRNAs), and is suggestive of an important role in spermatogenesis. PMID- 18390185 TI - [Purification and clinical application of egg yolk immunoglobulins]. AB - IgY antibodies, also called egg yolk immunoglobulins, are the only immunoglobulins in egg yolk and transferred in the female from serum to egg yolk to confer passive immunity to embryos and neonates. Using hens instead of mammals as the immunization host brings a number of advantages: Eggs are cheap and readily available; antibody levels in yolks are high; IgY isolation is fast and simple; and what is more; IgY neither binds the rheumatoid factor nor reacts to the mammalian complement factor. All these differences make IgY technology more widely applicable, such as in the production of polyclonal antibodies against various antigens, immunodiagnostics and immunotherapy, and many medical areas in both animals and human. IgY also has a good prospect in human immunocontraception. PMID- 18390186 TI - [Epimedium alleviates chemotherapy-induced damage to the ultrastructure and function of rat epididymides]. AB - OBJECTIVE: To investigate the protective action of Epimedium against chemotherapy induced damage to rat epididymides. METHODS: Fifty 60-day-old male rats were divided into a control, a model and a treatment group. Procarbazine was injected into the abdominal cavity of the model rats at the dose of 30 mg/(kg x d). In addition to procarbazine, Epimedium was given intragastrically to the treatment group. The changes in the ultrastructure of the epididymis were observed after 10 and 20 days. RESULTS: Electron microscopy showed that the chemotherapy-induced damages to the epididymal epithelia mainly included cell swelling, local cavitation of mitochondria, tumor-like change in nucleoli, agglutination of marginal translocation of heterochromatin and cell apoptosis. The damage to the epithelial ultrastructure was slight in the treatment group as compared with the model rats. Chemotherapy significantly affected sperm concentration, sperm viability and sialic acid (SA), which were (15.59 +/- 4.01) x 10(6)/ml, (76.71 +/ 10.11)% and (19.38 +/- 9.34) g/mg prot in the model group in comparison with (10.63 +/- 3.82) x 10(6)/ml (P < 0.01), (60.03 +/- 7.54)% (P < 0.01) and (13.62 +/- 7.81) g/g prot (P < 0.05) in the control. Epimedium significantly increased sperm viability in the treatment group (60.03 +/- 7.54)% as compared with the model rats (69.90 +/- 12.58)% (P < 0.05). CONCLUSION: Epimedium can lessen chemotherapy-induced damage to the epididymis and protect the reproductive function of rats. PMID- 18390187 TI - [Effect of aurora A on carcinogenesis of human prostate cancer]. AB - OBJECTIVE: To investigate whether Aurora A is involved in prostate cancer carcinogenesis. METHODS: The expressions of Aurora A mRNA and protein in prostate cancer tissue and cell line were measured using RT-PCR, immunohistochemistry and Western blot analyses. Meanwhile, A DNAzyme targeting at Aurora A mRNA was performed to detect the inhibition effect of Aurora A in regulating the growth of prostate cancer cell line PC3. RESULTS: Aurora A expression was up-regulated in 91% prostate cancer tissue as well as in prostate cancer cell lines such as PC3, LNCaP and Du145. Furthermore, Aurora A expression suppressed by cell transduction of a DNAzyme targeting at Aurora A mRNA made the PC3 cells both arrest the cell cycle in G2/M phase and increase the cell apoptosis. CONCLUSIONS: These findings suggest that Aurora A up-regulation may be involved in carcinogenesis of prostate cancer, and that aurora A gene may be a valuable target for treatment of prostate cancer. PMID- 18390188 TI - Long-term coculture of spermatogonial stem cells on sertoli cells feeder layer in vitro. AB - OBJECTIVE: To construct a cell model of spermatogonial stem cells (SSCs) cocultured on Sertoli cells feeder layer in vitro, and study the proliferation characteristics of SSCs. METHODS: Sertoli cells and SSCs were separated from testes of 14-15 days and 6 days KM mice respectively by two-step enzyme digestion. SSCs were seeded on the Sertoli cells layer at 5 days in culture. The clones of SSCs on the sertoli cells layer were detected, and cast-off cells in culture medium were counted. RESULTS: SSCs began to proliferate and differentiate 24 hours after being cultured on the Sertoli cells layer, and there were a few paired (Ap) cell clones. With more time of culture, the number of Ap cell clones decreased gradually, meanwhile the number of aligned (Aal) cell clones increased, then Aal cell colonies retained stable quantity after 120 hours in culture, it could retain (51.2 +/- 5.8) days under the condition of the culture medium being changed every 4 or 5 days. CONCLUSION: SSCs can proliferate colonially on the feeder layer of Sertoli cells, and retain stable morphous and quantity. SSCs cultured on Sertoli cell feeder layer provide a cell model for studying SSCs biological behavior and interruptions of drugs or toxins on spermatogenesis in vitro. Coculture PMID- 18390189 TI - [Effect of siRNA inhibiting HPV16 E6 gene expression on proliferation and cell cycle of cervical cancer cell line CaSki]. AB - OBJECTIVE: To observe whether human papillomavirus 16 (HPV16) E6-specific small interfering RNAs (siRNAs) can be employed to inhibit the growth of cervical cancer cell line, and to investigate the associated mechanism. METHODS: RNAi was performed using synthetic small interfering RNAs transferred into CaSki cell line by lipofectamine. The cell growth curves, live cell ratio and inhibition ratio of cells were measured by using cell counting. At various time points of post transfection, the distributions of cell cycle, the expression levels of HPV16 E6, p53, p21 mRNA and proteins were detected by using flow cytometry (FCM) and real time quantitative reverse transcription-polymerase chain reaction (real-time RT PCR). RESULTS: The growth inhibition of E6 siRNA to CaSki cells was demonstrated after cells treated with E6 siRNA. No substantial G1 arrest was observed by FCM analysis. For 24 hours after cell transfection, the level of E6 mRNA was decreased by 20. 11 folds compared with control (P < 0.05). However, p53 and p21 mRNA levels appeared unaffected. 48 hours after cell transfection, the expression level of E6 protein was efficiently decreased, but the P53 and P21 protein levels increased in comparison. CONCLUSIONS: The inhibitory effect of HPV16 E6 siRNA to CaSki cell maybe due to specially and efficiently silence E6 mRNA expression, decrease the degradation of wild type P53 protein, and then recover the function activity of P53 protein. PMID- 18390190 TI - [The expressions and distribution characters of glutamate receptor subunits NR1 and GluR2 in the hippocampal formation of adult rats]. AB - OBJECTIVE: To investigate the expressions and distributions of N-methyl-D aspartic acid NMDA/ NR1 and alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid AMPA/GluR2 subunits in the hippocampal formation. METHODS: The immunofluorescence histochemical staining was made on paraformaldehyde fixed hippocampal tissue sections for then observed by confocal laser scanning microscopy. RESULTS: There were widely positive expressions and basically same distributions of NR1 and GluR2 in the hippocampal formation of adult rats, especially in pyramidal layer of CA1 and CA3 regions and in the interneurons of hilus fascia dentata. Positive expressions, which mainly existed in the cytomembrane and processes of the neurons, also distributed in molecular layer of CA1 and CA3 regions as well as in granule layer of dentate gyrus. However, the immunoreactive intensity of NMDA/NR1 was stronger than that of AMPA/GluR2 in molecular layer of CA3 area and in polymorphic layer of dentate gyrus. CONCLUSION: The distributions of NMDA/NR1 and AMPA/GluR2 in the hippocampal formation are basically same, this suggests that they may execute cooperatively some important tasks. PMID- 18390191 TI - [Soluble expression, purification of bone morphogenetic protein-2 fragment and its antibody preparation]. AB - The bone morphogenetic protein-2 (BMP-2) fragment (BMP-2omega, 606-846bp) cDNA was amplified from total RNA of SAOS-2 cells by using RT-PCR. The PCR product was then inserted into pET-28a (+) vector for constructing the expression plasmid that would be used to transform the host cell BL21(DE3). After IPTG inducing under different conditions, this BMP-2w protein could be expressed in high level as a soluble form, and purified by chelating column (Ni-NTA). Polyclonal antibody was made by immunizing mice with using purified protein, and the antiserum titer generated was 1: 6400 that was measured by ELISA. Western blot result showed that this antibody could bind to BMP-2 protein specifically. Above research result set up the basis for studying on the treatment of osteosarcoma. PMID- 18390192 TI - [The polymorphism of interferon gamma gene CA short tandem repeat is associated with aplastic anemia]. AB - OBJECTIVE: To explore the possible relationship between the polymorphism of CA short tandem repeat in first intron of interferon-gamma (IFN-gamma) gene and the susceptibility of aplastic anemia. METHODS: 54 patients, who were diagnosed as acquired aplastic anemia in West China Hospital of Sichuan University when it was from February 2006 to March 2007, and simultaneously 51 healthy adults were enrolled as normal control for this project. The polymerase chain reaction and polyacrylamide gel electrophoresis were used to assay the polymorphism of CA short tandem repeat. RESULTS: The frequency of the homozygous for 12-12 (CA) repeats or the single allele 12 (CA) repeats of the patients was 18.52% or 50.92% respectively, which was obviously higher than the control group's frequency which was 2.00% or 26.47% (P = 0.008, P < 0.001 respectively). The frequency of the homozygous for 12 (CA) repeats or the single allele 12 (CA) in acute aplastic anemia group was 12. 00% or 48.00%, while in the chronic aplastic anemia group that frequency was 24.14% or 53.45%, the statistic analysis showed there was no significant difference between those two groups (chi2 = 1.311, P = 0.252, chi2 = 0.319, P = 0.572). CONCLUSION: This study suggests that: IFN-gamma CA short tandem repeat polymorphism is associated with the susceptibility of aplastic anemia but has no relation to the severity of the aplastic anemia. PMID- 18390193 TI - [Isolation of multipotent mesenchymal stem cells from the tissue of umbilical cord for osteoblasts and adipocytes differentiation]. AB - OBJECTIVE: To explore the methods and conditions for isolating and proliferating multipotent mesenchymal stem cells (MSCs) from the tissue of umbilical cord, with an aim to induce osteogenic and adipogenic differentiation in vitro. METHOD: The cord was dissected along the long axis, with vessels pulled away and then sutured into a "loop". Collagenase solution was filled into the loop. Suspended cells were collected from the loop suspension after 6-8 hours and centrifuged. The cells were finally cultured in polystyrene dishes. The single cell-derived colonies were obtained and tested for their immunophenotype and osteoblast and lipoblast differentiations. RESULT: Adherent cells were obtained from the tissue of umbilical cord, which proliferated and formed single cell-derived colonies. The colonies presented matrix cells immunophenotype and differentiated into osteoblasts that produced mineralized matrices, which were stained by alizarin red and alkaline phosphatase. The colonies also differentiated into adipocytes that accumulated lipid vacuoles, which were demonstrated by the morphology and oil red stains. CONCLUSION: MSCs can be isolate from the tissue of umbilical cords and proliferate in vitro. The proliferated colonies show matrix cell immunophenotypes and can differentiate into osteoblasts and adipocytes. PMID- 18390194 TI - [Relation between beta-amyloid peptide-25-35 inducing cell cycle change and apoptosis of serum-deprived PC12 cell]. AB - OBJECTIVE: To study the relation between cell cycle change and apoptosis of serum deprived PC12 cell, which are induced by beta-amyloid peptide-25-35 (alpha beta(25-35)). METHODS: PC12 cells were synchronized by cultured in deprivation of serum for 24 h and treated with different concentration of alpha beta(25-35) (0 45 micromol/L) for another 24 h,and the cell survival rate was evaluated by MTT assay. The cell apoptosis was analyzed by Hoechst fluorescence staining and DNA agarose gel electrophoresis. The relation between cell cycle redistribution and apoptosis was analyzed by flow cytometry (FCM). RESULTS: alpha beta(25-35) decreased the survival rate of PC12 cells in dose-dependent manner. The typical apoptotic cells were showed by fluorescence staining when treated with 25 micromol/L alpha beta(25-35) for 24 h;the obvious DNA-Ladder was showed by DNA agarose electrophoresis. About 90% PC12 cells were found to arrest on G0/G1 by FCM being deprived serum. Treated with 25 micromol/L alpha beta(25-35) for 8, 16, 24 h, the percent of S phase cells was raised remarkably (P < 0.01) at 8 h, but the percent of S phase cells was declined gradually after treated for 16 h. Meanwhile the apoptotic rate was detected being increased obviously between 16 h and 24 h (P < 0.01), the obvious hypodiploid peak could be observed ahead of G0/G1 phase(Ap). CONCLUSION: alpha beta(25-35) decreases the survival rate of synchronized PC12 cell and induces the synchronized PC12 cells attempting to reenter cell cycle,which appear the apoptotic peak subsequently. This indicates that the cell apoptosis may be related to the abnormal cell cycle distribution induced by alpha beta(25-35), which means there may be a close relationship between cell cycle and apoptosis. PMID- 18390195 TI - [Role of MMP-9 and TIMP-1 in renal tubular epithelial-myofibroblast transdifferentiation]. AB - OBJECTIVE: To investigate the dynamic changes of MMP-9 and TIMP-1 expressions in rat unilateral ureteral obstruction (UUO) model, and to explore the role of MMP-9 and TIMP-1 in tubular epithelialmyofibroblast transdifferentiation. METHODS: Sixty-four male rats were randomly allocated to sham UUO group and UUO group. Rats were sacrificed respectively at 3, 7, 14 and 21 days after UUO or sham surgery. The expression levels of MMP-9, TIMP-1, alpha-smooth muscle actin (alpha SMA) and fibronectin (FN) were examined by immunohistochemistry staining at each time point. RESULTS: In UUO rats, the disruption of renal tubular basement membrane (TBM) in PAS staining was found at day 3 after operation. At postoperative day 14 and 21, the TBM was observed to be thickened and wrinkled with partial and extensive disruption. On day 3 after UUO, the expressions of alpha-SMA and FN in tubulointerstitium were obviously increased. On day 7, some tubular epithelial cells also expressed alpha-SMA. In UUO rats, the strong positive expression of MMP-9 in renal tubulointerstitium was found at day 3 after operation and the expression peaked in tubulointerstitium at day 7, then decreased gradually. (4) In UUO rats, TIMP-1 expression in renal tubulointerstitium was increased progressively starting from day 3 to day 21. (5) The expression of MMP-9 in renal tubulointerstitium was significantly associated with the positive area of alpha-SMA and the disruption of TBM. CONCLUSION: The expression levels of MMP-9 and TIMP-1 relate closely to the disruption of TBM. TBM structural and functional integrity depends on many intricate interactions between MMP-9 and TIMP-1, which may play an important role in tubular epithelialmyofibroblast transdifferentiation. PMID- 18390196 TI - [Brain structural differences of schizophrenia patients with and without family histories]. AB - OBJECTIVE: To identify the brain structural differences of schizophrenia patients and their parents between those with family history and those sporadic cases. METHODS: High resolution T1 weighted images were obtained using 3T MR from 10 familial patients and 10 sporadic patients matched with age, sex, years of education and duration of disease. All patients were in their first episode of care. Twenty unaffected parents of the patients (8 for familial cases, 12 for sporadic cases) and 20 normal controls were also examined. The images were preprocessed according to the optimized VBM protocol. Student t test was performed to test the differences of the gray matter density (GMD) of the patients and their parents between the groups with and without family histories, and between the patient groups and the normal controls. RESULTS: Compared with the sporadic group, significant reduce of GMD was observed in bilateral thalamus in both familial schizophrenia patients and their parents. Compared with normal controls, both familial and sporadic schizophrenia patients showed lower GMD in many areas including bilateral insula, right temporal lobe, right occipital lobe, left lenticula, right cerebellum and left rectal gyrus extending to anterior cingulated gyrus. Only familial patients showed lower GMD in the right thalamus. Familial parents also showed lower GMD in several areas including right insula extending to right temporal lobe and right parietal lobule. CONCLUSION: Familial schizophrenia is associated with genetically related structural abnormalities, especially in the bilateral thalamus. The brain structural differences can explain, at least in part, the symptom differences between familial and sporadic schizophrenia. PMID- 18390197 TI - [The human-mouse chimera nude mouse model set up from umbilical cord blood transplantation]. AB - OBJECTIVE: This study was to establish BALB/c murine model featured with the human-mouse chimeras from umbilical cord blood transplantation. METHODS: Thirty BALB/c nude mice were exposed to 350 cGy radiation under the sublethal condition. The nuclear cells from fresh umbilical cord blood were injected into the mice of experimental group via tail vein, in which the mice were further allocated to A, B and C sub-group with given 1.0 x 10(7), 2.0 x 10(7) and 3.0 x 10(7) nuclear cells per mouse respectively, and simultaneously the equal volume normal sodium (NS) was injected into the mice of control group. White blood cells from peripheral blood in experimental group and the control group were detected at week 1, 2, 3, 4 and 8 after transplantation. Human CD34+ and CD45+ from peripheral blood in experimental group were detected by using flow cytometry analysis on week 4, 6 and 8 after transplantation in order to be the human-mouse chimeras known. RESULTS: No difference in the number of white blood cells showed between experimental groups and control group before blood cell transplantation and after transplantation conducted on for 8 weeks (P > 0.05), but the number of white blood cells from experimental groups and control group wasn't totally same on week 1, 2, 3 and 4 after cell transplantation conducted (P < 0.05). As compared with pre-transplantation, the number of white blood cells for post transplantation at different time in experimental groups and control group was decreasing, and the lowest on 1 week, then going up and to the level of pre transplantation on 4 week in experimental groups and on 8 week in control group. CD34+ and CD45+ cells in peripheral blood for nude mice appeared on 4 week, but the total number was little. The number of CD34+, CD45+ cells in experimental group A was less than those in experimental group B and C at all the stage (P < 0.05), but there was no difference between experimental group B and C(P > 0.05). There was obvious correlation between CD34+ and CD45+ on 4 week (r = 0. 903, P < 0.05), but this correlation didn't appear on 6 and 8 week (P > 0.05), which might be related to the amount of sample. CONCLUSION: The BALB/c murine model can be successfully established with the human-mouse chimeras from umbilical cord blood transplantation. PMID- 18390198 TI - [Expression of Sema3A and NP-1 in spinal cord and spared dorsal root ganglion after partial dorsal root rhizotomy of cat]. AB - OBJECTIVE: To investigate the expression of semaphorin 3A (Sema3A) and neuropilin 1 (NP-1) in spinal cord and dorsal root ganglion after partial dorsal root rhizotomy. METHODS: 15 adult cats were used for this study and divided into 3 groups: normal control group, 7 d and 14 d postoperative groups (7Th day and 14th day groups) undergoing unilateral partial dorsal root rhizotomy. The L3, L5 and L6 segments of spinal cord and L6 dorsal root ganglia (DRG) in operated side were made into frozen sections. By immunohistochemistry ABC method, the sections of spinal cord were stained with specific Sema3A antibody, and L6 DRG were stained with NP-1 antibody. The mean optical density (OD) of Sema3A immunoreactivity in dorsal horn was measured and the number of NP-1 positive medium-small sized neurons in spared DRG was counted. RESULTS: After partial dorsal root rhizotomy, in L3 segment the expression of Sema3A decreased in 7th day group (0.25 +/- 0. 14) compared with that in normal group(0. 37 +/- 0.87) (P < 0.05), but kept the level along to 14th day group (0.27 +/- 0.09); in L5 segment, the expression of Sema3A decrea sed in 7th day group (0.26 +/- 0.11) (P < 0.05), and then recovered to normal level in 14th day group (0.33 +/- 0.09); in L6 segment, OD values in dorsal horn had no changes to all groups. The number of NP-1 positive medium small sized neurons in spared DRG (30.85 +/- 10.26) was decreased in 7th day group (P < 0.05), compared with that in normal group (45.06 +/- 12.47), while increased in 14th day group (P < 0.05). CONCLUSION: Changes in the expression of Sema3A in spinal cord and the expression of NP-1 in L6 DRG after partial root rhizotomy may be involved in collateral sprouting of spared root in superficial lamina. PMID- 18390199 TI - [Correlation between lymph-node metastasis and combined expression of Hpa and C erbB-2 of invasive ductal breast carcinoma]. AB - OBJECTIVE: This study is to learn the correlation between lymph-node metastasis and the combined expression of Heparanase (Hpa) and C-erbB-2. METHODS: The expressions of Hpa and C-erbB-2 were examined in 81 human invasive ductal breast carcinomas ( IDBC ) and 14 fibroadenomas by SP immunohistochemistry analysis. We analyzed the expressions of two factors in two groups, in which their correlation with lymph-node metastasis in IDBC was also studied. RESULTS: (1) Positive expressions of Hpa and C-erbB-2 in IDBC tissue were 59 (72.84%) and 29(35. 80%) respectively, in which the positive expression rates were significantly higher than those in fibroadenomas (P = 0.0000, 0.0177). (2) The expression of C-erbB-2 was correlated with Hpa expression (r = 0.668, P < 0.01). (3) When Hpa expression was combined with the status of C-erbB-2 expression, the lymph node metastasis with positive-Hpa and positive-C-erbB-2 expressions occurred significantly higher in patients with IDBC (all P < 0.01). CONCLUSION: Hpa may up-regulate the expression of C-erbB-2. Furthermore, as co-expression, Hpa and C-erbB-2 work together, which would take the possession of stronger role in the procedure of growth, invasion and metastasis of IDBC. PMID- 18390200 TI - [Study on genetic polymorphisms of P-selectins in Chinese Han of Chengdu and Thai populations]. AB - OBJECTIVE: To study the gene polymorphisms of position --2123 C/G,--1969 G/A,- 1817 T/C in promoter region and of Thr715Pro in exon thirteenth of P-selectin in the Chinese Han of Chengdu and Thai populations, and simultaneously to compare distributions of genotype and allelic frequencies of P-selectins among different races. Methods The genotypes and allele frequencies of the P-selectin base --2123 C/G,--1969 G/A,-1817 T/C and amino acid Thr715Pro were detected by polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) to 120 healthy Chinese Han of Chengdu and 110 Thai population. RESULTS: There were no significant differences in the genotype and allele distribution of--2123 C/G,- 1969 G/A,--1817 T/C polymorphisms for the P-selectin gene between Chinese Han of Chengdu and Thai populations (P > 0.05), in which compared with England and American, the distribution of P-selectin genotype and allele had significantly differences among ethnics (P < 0.001). No polymorphism of Thr715Pro was found in this study. Conclusion In Chinese Han of Chengdu and Thai populations the polymorphisms exist at base position--2123 C/G,--1969 G/A and --1817 T/C in promoter region of P-selectin. There are no significant differences in the genotype and allele distribution of the P-selectin gene polymorphisms between Chinese Han of Chengdu and Thai populations, but significantly different distribution of P-selectin gene polymorphisms occur among ethnics. PMID- 18390201 TI - [Genetic polymorphisms of five STR gene loci GATA30F04, GATA23B10, D18S847, GATA83B04 and GATA167A05 alleles in Chinese Han population of Chengdu area]. AB - OBJECTIVE: To get the preliminary genotype and allele frequency distributions of GATA30F04, GATA23B10, D18S847, GATA83B04 and GATA167A05 loci in Chinese Han population in Chengdu area, and to validate more short tandem repeat (STR) systems for forensic application. METHODS: The polymerase chain reaction (PCR) was used to this study. Five STRs (GATA30F04, GATA23B10, D18S847, GATA83B04 and GATA167A05) were amplified from DNA samples, which were extracted with Chelex-100 method from EDTA-blood of 100 unrelated individuals. The PCR products were analyzed by PAG vertical electrophoresis. RESULTS: No deviations from Hardy Weinberg equilibrium were observed. The expected heterozygosities observed were 0.694, 0.918, 0.836, 0.889 and 0.880 for GATA30F04, GATA23B10, D18S847, GATA83B04 and GATA167A05 respectively. The discriminating powers were 0.697, 0.901, 0.875, 0.900 and 0.891. CONCLUSION: The last four loci in this study are useful markers for genetics purposes in individual identification and paternity test. PMID- 18390202 TI - [Effect of Janus kinase inhibitor AG490 on invasion and metastasis of human breast cancer cells]. AB - OBJECTIVE: To investigate the effect of Janus kinase (JAK) inhibitor AG490 on invasion and metastasis of the human breast cancer cell MDA-MB-231, and to explore the regulating role of JAK-STAT3 signaling pathway when the breast cancer occurs to the invasiveness and metastasis. METHODS: The human breast cancer cell MDA-MB-231 was used as the research object, and AG490 was as Janus kinase inhibitor. The adhesion of MDA-MB-231 cell attaching to matrigel was measured with MTT assay. The invasion and metastasis potential were evaluated with transwell chamber. The P-STAT3 protein in cell was detected by Western-blot. RESULTS: Janus kinase inhibitor AG490 could make that the expression of P-STAT3 in human breast cancer cell MDA-MB-231 became weak, and that the abilities of adhesion, invasion and metastasis were also dropping down as compared with the control group (P < 0.01). CONCLUSION: JAK-STAT3 signaling pathway participates in regulating the invasion and metastasis of human breast cancer. Inhibiting the activation of JAK-STAT3 signaling pathway can suppress the invasion and metastasis of human breast cancer. PMID- 18390203 TI - [The change of glucagon-like peptide-1 and its effect on blood glucose metabolism after major surgery]. AB - OBJECTIVE: To investigate the change of glucagon-like peptide-1(GLP-1) and its effect on blood glucose metabolism after major surgery. METHODS: Eleven patients, who had undergone major surgical procedures in our Department of General Surgery, were studied on the day before surgery, the first, third, and fifth day after surgery. Then, 42 rats were allocated randomly into three groups. The rats in control, which had not undergone any operation, received an intravenous glucose load (0.5 g/kg glucose + normal saline). The rats in operated group, which had undergone about 65% liver resection, received a same glucose load on the first, third,and fifth postoperative day. And the rats in GLP-1 group, which had undergone same hepatectomy, received a same glucose load with GLP-1 (0.3 nmol/kg) on the first, third,and fifth day after surgery. All rats would be killed by abdominal aorta exsanguinated in five minutes after drugs were infused of which the bloods were collected for determination of glucose (glucose oxidase), insulin, glucagon, and GLP-1 (radioimmunoassays) at each time-point. RESULTS: There was an increasing postoperative plasma concentration of glucose, insulin, and glucagon on the first day (P < 0.01), but the plasma GLP-1 was just elevated on the third day (P < 0.05). Then, the plasma glucose concentration was significantly lowered after GLP-1 given to rats undergoing hepatectomy (P < 0.001), which might reach the glucose range in controls. Lowering of blood glucose was achieved by a significant rise of insulin secretion (P < 0.001) and a suppression of glucagon secretion (P < 0.05). CONCLUSION: As far as can be concluded on basis of our data from patients and rats, GLP-1 can be used to reduce the plasma glucose concentrations when it is in stress status after major surgery. PMID- 18390204 TI - [Damage mechanism of intermittent exposure to high concentrations of glucose to beta cell lines (HIT-T15 cell)]. AB - OBJECTIVE: To test the effect of intermittent exposure to high concentrations of glucose on HIT-T15 cells. METHODS: The HIT-T15 cells (pancreatic beta cell lines) were exposed to 16.7 mmol/L of glucose for two hours, at an interval of three hours. Such intermittent exposure was repeated three times a day. Otherwise, the cells were exposed to 5.5 mmol/L of glucose. The control groups included continuing exposure to 5.5 mmol/L of glucose (normal control group) and continuing exposure to 16.7 mmol/L of glucose. The impact of N-acetyl-L-cysteine (NAC) was also tested in the groups with intermittent and continuing exposure to 16. 7 mmol/L of glucose. The insulin was detected by radioimmunoassay (RIA). The mitochondrial membrane potential (MMP) was measured by laser scanning confocal microscope. The ATP levels of beta cells were examined by the luciferin luciferase kit. The Malonaldehyde (MDA) was measured through thiobarbituric acid reaction (TBA). The intracellular reactive oxygen species (ROS) was observed by flow cytometry. RESULTS: The HIT-T15 cells exposed to intermittent and continuing high glucose secreted [(3.13 +/- 1.11) microIU/mLD and [(5.18 +/- 0. 95) microIU/mL] of insulin, significantly less than the control [(9.33 +/- 0.62) microIU/mL, P < 0.05]. The MDA and ROS levels increased while the ATP and MMP levels decreased significantly in the cells exposed to intermittent and continuing high glucose (P < 0.05). The cells exposed to intermittent high glucose produced more ROS than the cells exposed to continuing high glucose (P < 0.05). CONCLUSION: Exposure of HIT-T15 to high glucose leads to oxidative stress of mitochondrial. Intermittent exposure causes more serious stress than continuing exposure. Such response might be a result of decreased MMP level and ATP contents. PMID- 18390205 TI - [Effect of simvastatin on endothelin-1 expression in endothelial cell cultured hypoxically]. AB - OBJECTIVE: To observe the role of simvasatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, in synthesis and excretion of endothelin-1 (ET-1) in endothelial cell cultured hypoxically. METHODS: Human umbilical vein endothelial cells were hypoxically cultured and treated with simvastatin by different concentrations (0, 1.0, 2.5, 5.0, 10.0 micromol/L) and different times (12, 24, 48 h). Mevalonate was used to intervent the effect of simvastatin. Reverse transcription-polymerase chain reaction (RT-PCR)and enzyme-linked immunoadsorbent assay (ELISA) were adopted to measure ET-1 mRNA and ET-1 in supernatant fluid of endothelial cell culture. RESULTS: (1) There were no changes of ET-1 mRNA and ET-1 expression after the hypoxically cultured endothelial cell were incubated with 1 MICROmol/L simvastatin, but ET-1 expression decreased without significant difference compared to control (0 micromol/L simvastatin) when interfered with 2.5 micromol/L simvastatin. The decreases of ET-1 mRNA and ET-1 expression became more obvious when expression were interfered by 5 micromol/Land 10 micromol/L simvastatin (P < 0.01). (2) ET-1 mRNA and ET-1 expression decreased at 12 h after the endothelial cells were incubated with 10 micromol/L simvastatin, which became more fewer at 24 h and reached the minimum expression at 48 h (P < 0.01). (3) The inhibition effect of simvastatin on ET-1 mRNA and ET-1 expression of endothelial cells could be prevented by mevalonate with concentration of 100 micromol/L. CONCLUSION: Simvastatin can inhibit ET-1 expression in endothelial cell cultured hypoxically. PMID- 18390206 TI - [Experimental study in renal protective effect of tranilast on rats with adriamycin nephropathy]. AB - OBJECTIVE: To investigate the effect and mechanism of tranilast on the adriamycin induced nephrotic syndrome of rats. METHODS: Twenty four rats were randomly divided into 4 groups: normal control group, model group, irbesartan treatment group and tranilast treatment group. The rats in normal control group were injected with normal saline via the tail vein. The rats in the other groups were uninephrectomized and injected with adriamycin 5 mg/kg via the tail vein one week later. Rats in model group underwent gavage to receive the sodium carboxymethylcellulose, rats in irbesartan treatment group to receive the irbesartan with 10 mg/kg x d, and rats in tranilast treatment group to receive the tranilast with 400 mg/kg. Rats were then sacrificed at the end of week 8. The body weight, 24 hours urinary protein, blood urea nitrogen (BUN) and serum creatinine (Scr) of each group rats were measured for the study. Renal pathological changes were evaluated. And immunohistochemistry was used to examine the expression of transforming growth factor beta1 (TGF-beta1) and tissue inhibitor of metalloproteinase 1 (TIMP-1). In situ hybridization was used to measure the expression of a-smooth muscle actin (alpha-SMA) mRNA in the kidney. RESULTS: Compared with the untreated nephrotic syndrome rats, the proteinuria and Scr of rats treated with tranilast were significantly reduced (P < 0.05); Compared with model group, the renal pathological changes of rats in tranilast treatment group were decreased, with glomerular sclerosis to be markedly lower; Tranilast could decrease the expression of TGF-beta1, TIMP-1 and alpha-SMA mRNA in the kidney of rats with adriamycin nephropathy. CONCLUSIONS: Tranilast has a renoprotective effect on adriamycin-induced nephrotic syndrome in rats, of which the mechanism may be related to that tranilast can depress the expression of TGF beta1, and TIMP-1 in the kidney, with result in decreasing the synthesis and secretion of extracellular matrix. And tranilast inhibits the transdifferentation of renal primary cells, regulates the synthesis and degradation system of extracellular matrix. PMID- 18390207 TI - [Coriaria lactone induces in vitro the overexpression of multidrug resistance P glycoprotein in astrocyte from rat brain]. AB - OBJECTIVE: The aim of this study was to explore the possible pharmacoresistant mechanism by developing a kind of astrocyte model induced by Coriaria lactone (CL). METHODS: Astrocyte cells from natal Sprague-Dawley rats (within 24 hours of birth) were cultured. Different volumes of CL (1 microL, 3 microL, 9 microL, 15 microL) were added to the cultures for 7, 14, 21 or 28 days. The expression of P glycoprotein (Pgp), the protein product of multidrug resistance 1 (MDR1), was investigated with immunocytochemistry and image analysis. RESULTS: CL induced the overexpression of Pgp in astrocytes in a time-dependent manner. The up-regulation of Pgp expression was detected to any concentration of CL from day 14 to 28 (P < 0.01). On the day 21, 28, the up-regulation of Pgp expression level was no dose dependent manner (P > 0.05). CONCLUSION: The study provides a ideal astrocyte model of pharmacoresistant epilepsy, which is induced by CL. The increased expressions of P-gp may involve in induction of the pharmacoresistance. PMID- 18390208 TI - [Effects of dl-3-n-butylphthalide on expression of VEGF and bFGF in rat brain with permanent focal cerebral ischemia]. AB - OBJECTIVE: To study the effects of dl-3n-butylphthalide (NBP) on the protein and mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in rats brain with permanent middle cerebral artery occlusion (MCAO). METHODS: The model of permanent MCAO was established by using the suture method of Longa, with which the nylon suture was used to make rat middle cerebral artery (MCA) blocked. Sham-operated rats (n=20) were prepared in similar fashion, but without doing the closed occlusion of the MCA. Operated rats were randomizely divided into model control and NBP groups (n= 20 for each). By intragastric administration, sham-operated and model control group rats were given vegetable oil 2 mL twice daily for 3 days, and also NBP group rats were given NBP 25 mg/kg twice daily for 3 days. The infarct volume and neurological deficit scores were determined by tetrazolium chloride (TTC) staining and Longa's score separately. The protein and mRNA of VEGF and bFGF were detected by immunohistochemistry and in situ hybridization. RESULTS: NBP markedly inhibited the neurological deficit and reduced the infarct volumes as compared to model control group (P < 0.05). NBP significantly upregulated VEGF and bFGF expression in both protein and mRNA levels in the peripheral infarct and hippocampus regions in contrast with sham-operated and model control groups (P < 0.05). In the infarct core, the protein and mRNA levels of VEGF and bFGF did not show significantly any difference (P > 0.05). CONCLUSION: NBP can significantly reduce neurological deficit and infarction volume, and therefore may have protective effect for cerebral ischemia through upregulating the expression of VEGF and bFGF. PMID- 18390209 TI - [Effects of preemptively injected intrathecal lornoxicam on behavior and c-fos protein expression of rat with formalin hurting]. AB - OBJECTIVE: The objective of this study was to investigate an antinociceptive effect of preemptive intrathecal lornoxicam on behavior and expression of c-Fos protein in the spinal cord of the formalin hurting rats. METHODS: Forty-two healthy male adult SD rats (200-250 g) were randomized into 7 groups. In the normal saline control group (group NS), 20 microL of normal saline was injected intrathecally, which was followed by a subcutaneous administration of 100 microL of normal saline into the hind plantar skin of rats 10 min later. In the formalin control group (group FOR), 20 microL of normal saline was injected intrathecally, and followed by rat intraplantar administration of 100 microL of 5% formalin. In the intrathecal lornoxicam experiment groups (groups L60-300), lornoxicam 60, 120, 180, 240, and 300 micdrog were administered intrathecally, respectively, and followed by rat intra-plantar administration of 100 microL of 5% formalin. The graded pain behavior was observed and the nociceptive behavior index (NBI) was calculated per 5 min during one hour immediately after the intra-plantar injection. At the 2-hour time point, all the rats were deeply anesthetized and perfused intracardially; and then, the spinal cord was removed for analysis of c Fos protein-like immunoreactive neurons (FLIN) at the dorsal horn in the L4-L6 segments. The Student-Newman-Keuls test was used to identify the differences between groups,and correlation analysis was used to demonstrate the relationship between rat behavior and c-Fos expression. RESULTS: Almost no pain response was observed in group NS. The typical biphasic pain response was observed in the formalin-injected groups. In phase I, no difference in the NBI was found between the formalin-injected groups except group L240 and group L300 in which NBI were significantly smaller than that in other formalin-injected groups. In phase II, the cumulative NBI in phase II decreased with the increase in lornoxicam dosage, but the difference was not significant when the dosage above 180 microg. (2) Both in the superficial and deep laminae at the dorsal horn, the amount of FLIN of group NS was much less than that of the remaining groups. The amount of FLIN decreased with the increase in the lornoxicam dosage, but the difference was not significant when the dosage above 180 microg in the superficial lamina and above 240 microg in the deep lamina. (3) Correlation analysis showed that the correlation coefficient between the sum of NBI in one hour and the expression of c-Fos in the superfacial or deep lamina of dorsal horn was positive. CONCLUSION: Pre-intrathacal lornoxicam can successfully inhibit nociceptive behavior and c Fos expression in spinal dorsal horn of formalin test rats. This effect is correlated with the dosage of lornoxicam and has ceiling phenomenon. PMID- 18390210 TI - [Inhibitive effect of local perfusion of tanshinone II A nanoparticles on MMP-2 secretion]. AB - OBJECTIVE: To test the effect of Tanshinone II A nanoparticles on the expression of MMP-2. METHODS: Thirty five male New Zealand white rabbits were randomly divided into three groups. In group A (15 rabbits), the carotid arteries of the rabbits were stripped. In group B (5 rabbits) and C (15 rabbits), nanoparticles and Tanshinone II A nanoparticles were perfused respectively in the denudated arteries of the rabbits. The neointimal areas and the IOD values of MMP-2 secretion were measured. RESULTS: A significant reduction of neointimal hyperplasia in group C was found compared to the other two groups in terms of the intimal area and the intima-media ratio. Group C had significant lower IOD values than group A. CONCLUSION: Local administration of nanoparticles with incorporated Tanshinone I A not only inhibits neointimal hyperplasia but also inhibits the expression of MMP-2 in stripped arteries. PMID- 18390211 TI - [Time-dependent effects of pentabrominatefl diphenyl ethers on thyroid hormones and histological structure of thyroids in rats]. AB - OBJECTIVE: To study the time-dependent effects of pentabrominated diphenyl ethers on thyroid hormones and the histological structure of thyroids in rats. METHODS: One hundred healthy 4-week-old SD rats were randomly divided into 10 groups. The rats in the experimental groups were orally administered with pentabrominated diphenyl ethers (BDE-99) at a dose of 60 mg/kg for 10, 15, 20, 30 and 40 days respectively, while the rats in the control groups received equal volumes of corn oil for the same periods of time. Serum FT4, FT3 and TSH were measured by radioimmunoassay. Sections of thyroids were dyed with hematoxylin-eosin (HE) to detect pathological alterations in thyroid tissues. RESULTS: Decreased serum FT4 and FT3 and increased TSR were detected in the BDE-99 treated rats at DiS and D20. No significant differences of serum hormones were found between the experimental and control groups at D10, D30 and D40. The thyroid follicle epithelium of the BDE-99 treated rats proliferated into multiple-layer structure at DiS. At D20, proliferative plaque formed in local areas. Solid buds and follicles secondary to the solid buds were detected at D30 and D40. CONCLUSION: BDE-99 leads to structural changes of thyroids, reduces serum FT4 and FT3 and increases TSR. The time-dependent effects are evident in a damage and compensate pattern. PMID- 18390212 TI - [The expression rule of proliferating cell nuclear antigen in rat thyroid of hyperplasia induced by propylthiouracil]. AB - OBJECTIVE: To study the expression rule of proliferating cell nuclear antigen (PCNA) in thyroid when the hyperplasia of male rat thyroid is induced by propylthiouracil (PTU). METHODS: PTU was administered to rats by gavage at a dose of 5.0 mg/kg B. W for 0,3,6,9 and 12 days respectively. All animals were sacrificed after the last dosage, and the expression of PCNA in thyroid was detected by immunohistochemistry and RT-PCR. RESULTS: Compared with the control group, the PCNA level in the thyroid of rats given PTU for 3 days significantly increased (P < 0.05),and reached the highest expression when it was 6 days, but then when it was 9 days and 12 days, the PCNA expression showed a descending tendency. CONCLUSION: PCNA has a expression rule in hyperplastic thyroid: expression increase first but then decrease. This rule confirms the histological change of thyroid in earlier stage. The experimental time of assessment test to thyroid hormone disruptors will be shortened to six days. PMID- 18390213 TI - [Impact of anxiety-depression symptoms on clinical recovery and serum TNF-alpha and IL-6 of patients with burns]. AB - OBJECTIVE: To investigate the impact of anxiety-depression symptoms on the healing of burn wounds and the serum tumor necrosis factor a (TNF-alpha) and interleukin-6 (IL-6) in the patients with moderate and severe burns. METHODS: The patients with moderate and severe burns were divided into anxiety-depression group (24 patients) and non-anxiety-depression group (27 patients)according to the scores of anxiety-depression scores three days after burn measured by Hamilton Rating Scale for Depression (HAMD) and Hamilton Rating Scale for Anxiety (HAMA). The routine therapy was applied for both groups for 30 days. Then the scores of anxiety and depression and the degrees of wound healing were examined. The serum TNF-alpha IL-6 were measured with enzyme linked immunosorbent assay (ELISA). RESULTS: No significant decrease of the scores of depression was found before and after treatments in the anxiety-depression group (P > 0.05). The days that were needed for healing the wounds and the serum TNF-alpha and IL-6 increased in the patients with depression compared with the control group (P < 0.05). CONCLUSION: There are anxious and depressive reactions to burns by the patients. The continuous negative emotion can induce the increase of serum TNF alpha and IL-6 and lead to delay of the healing of the burn wounds. PMID- 18390214 TI - [Effect of propofol aortic infusion on ischemia-reperfusion spinal cords of rabbits]. AB - OBJECTIVE: To reveal the mechanism of propofol aortic infusion in protecting ischemia-reperfusion spinal cords through detecting the IL-6 levels in the spinal cords and observing the neurological functioning. METHODS: Fifty six healthy New Zealand White rabbits were randomly allocated into three groups, 4 in the control group, 26 in the Saline infusion group and 26 in the Propofol infusion group. The spinal cord ischemia was induced by infrarenal aorta occlusion for 30 minutes. Propofol were infused through aorta distal to the occlude sites of the rabbits in the Propofol infusion group continuously with a pump at a rate of 50 mg/kg x 30 min. The same volume of Saline were infused in the same way and at the same rate to the rabbits in the Saline infusion group. The lumbar segments of 4-6 spinal cords were harvested and the IL-6 were examined 0 hour or 2 hours after reperfusion. The spinal cords of the rabbits in the control group were harvested right after anesthesia. Forty eight hours after reperfusion, the neurological functioning of the rabbits was assessed with the Tarlov scale system and the normal motor neurons of anterior horn of the lumbar segments of 4-6 spinal cords were counted. RESULTS: The IL-6 levels of the rabbits in the Saline infusion group were significant higher than those in the control group and the Propofol infusion group (P < 0.05). There was no significant difference in the IL-6 level between the Propofol infusion group and the control group (P > 0.05). A significant increase of IL-6 in the rabbits in the Saline infusion group and the Propofol infusion group 2 hours after reperfusion was observed (P < 0.05). The rabbits in the Propofol infusion group had less paraplegia (30%) and more normal neurons (8.4) than those in the Saline infusion group (80% and 1.9, respectively) (P < 0.05). CONCLUSION: Occlusion of aorta increases IL-6 in the injured spinal cords. Propofol aortic infusion can decrease the IL-6 level and improve the neurological functioning, which is perhaps associated with the inflammatory inhibition effect of Propofol. PMID- 18390215 TI - [Effects of connective tissue growth factor on angiogenesis of random skin flaps in rats]. AB - OBJECTIVE: To investigate the effects of connective tissue growth factor (CTGF) on angiogenensis of random skin flaps in rats. METHODS: Thirty-two SD rats were randomly divided into three CTGF groups (rCTGF 120 ng, rCTGF 160 ng, rCTGF 200 ng) and one control group. Ischemic random dorsal skin flaps (2 cm x 8 cm) were constructed on the backs of the rats. In the CTGF groups, connective tissue growth factor (CTGF) was injected subcutaneously to the rats in the skin flaps at the time of surgery. The same volume of 0.9% NaCl solution was injected to the rats in the control group. Ten days later, the amount of viable tissues within the flaps were examined by planimetry. Specimens from the random skin flaps were harvested for immunohistological and HE staining to examine the microvascular growth. RESULTS: The immunohistochemical staining and HE staining revealed that the three CTGF groups had more vessels than the control group (21.00 +/- 3.423, 27.38 +/- 3.114 and 37.38 +/- 3.583 vs. 13.88 +/- 1.959, P < 0.05). Similar results were found in the middle parts of the skin flaps (15.50 +/- 2.777, 24.63 +/- 3.335 and 28.63 +/- 4.984 vs. 9.50 +/- 1.927, P < 0.05). CONCLUSION: CTGF may enhance the survival of skin flaps in a dose-dependent manner, which is associated with an increase in angiogenesis of skin flaps. PMID- 18390216 TI - [Polymorphism of estrogen receptor alpha genes Pvu I, Xba I and TA repeat sequence and hypomenorrhea with unknown aetiology]. AB - OBJECTIVE: To study the roles of the polymorphism of the estrogen receptor genes in hypomenorrhea with unknown aetiology. METHODS: A case control study was carried out in south west of China, with 100 patients with hypomenorrhea in the case group and 100 eumenorrhea women in the control group. Molecular biology test was undertaken to test the restriction fragment length polymorphism (RFLP) of the first intron incision enzyme Pvu II, Xba I in ERa gene. Depuration, clone and sequence analysis was performed to the TA repeated sequence in the hypervariable region of estrogen receptor gene. The genotype distribution of ERa gene polymorphism was compard between the case and control groups. RESULTS: The patients with hypomennorrhea had higher P genotypic frequency (47.5%) than the control (30.5%), with an OR of 1. 810 (95% CI = 1.113-2.765, P = 0.012). The patients with hypomennorrhea had lower X genotypic frequency (20.5%) than the control (32.0%), with an OR of 0.641 (95% CI = 0.361-0. 898, P = 0.036). The patients with hypomennorrhea had higher frequency of TA13 allele (P = 0.006) and lower Frequency of TA15 allele frequency (P = 0.033) than the control. CONCLUSION: ERa gene polymorphism is associated with hypomenorrhea with unknown aetiology. P allele and TA13 allele may be risk factors, while X allele and TA15 allele may be protective factors. PMID- 18390217 TI - [Relationship between cardiovascular complication and inflammation, oxidative stress of patient with maintenance hemodialysis]. AB - OBJECTIVE: To evaluate and analyze relationships between states of inflammation, oxidative stress and complicatedly cardiovascular diseases in patients with maintenance hemodialysis (MHD). METHODS: The analytical method for xanthine oxidase was used to measure the serum superoxide dismutase (SOD), spectrophotometry was used to measure the serum MDA, GSHPx. The immune turbidimetry was used to measure the serum levels of C-reactive protein (CRP) in patients with chronic renal failure. RESULTS: Compared with normal control group, the uremia groups had lower serum GSHPx, SOD and ALB levels, but higher CRP and MDA levels (P < 0.01). The MHD patients exhibited more serious disturbance of inflammation and oxidative stress which were aggravated as MHD time extended. CRP and MDA were significantly increased but GSHPx,SOD and ALB decreased in patients with cardiovascular events compared with the patients without them (P < 0.01). The results of Pearson correlation analysis showed: CRP was correlated with MDA positively (r = 0.740, P < 0.01), while with GSHPx, SOD, ALB negatively (r = 0.684, -0.688, -0.924, P < 0.01). ALB was correlated with GSHPx, SOD positively (r = 0.744, 0.728, P < 0.01), while with MDA negatively (r = -0 779, P < 0.01). The results of Logistic regression analysis showed that CRP contributed to the predictability of incidence of CVD (regression coefficient b = -1.054, Wald chi square = 5.251, OR = 0.348, P < 0.05). CONCLUSION: MHD patients are in the states of inflammation and oxidative stress which are aggravated with the prolonging of hemodialysis time. Inflammation and oxidative stress may be the risk factors for occurrence of cardiovascular events. PMID- 18390218 TI - [Study on levels of angiotensin converting enzyme and C-reactive protein in peripheral blood of mitral valve replacement preoperative patients with atrial fibrillation]. AB - OBJECTIVE: To evaluate the levels of angiotensin converting enzyme and C-reactive protein in the peripheral blood of mitral valve replacement preoperative patients with atrial fibrillation (AF). METHODS: 24 atrial fibrillation patients, who were hospitalized for mitral valve replacement surgery, as study group (AF group) and 23 sinus rhythm patients as control group (SR group) were included in this study. The angiotensin converting enzyme and C-reactive protein in peripheral blood were quantified and compared in an attempt to find out whether the significant difference existed between the AF and SR groups. All patients undertook ultrasonic cardiography before operation. RESULTS: There was no significant difference in C-reactive protein between two groups (P > 0.05). The quantity of angiotensin converting enzyme in patients with AF was higher than that in patients with SR, which was statistically significant (P < 0.05). And also the left atrial dimension had a linear correlation with angiotensin converting enzyme. The obvious relation between angiotensin converting enzyme and left atrial dimension was seen in AF patients who need mitral valve replacement. CONCLUSIONS: This study demonstrates that the increase of the quantity of angiotensin converting enzyme in peripheral blood has relationship with atrial fibrillation of patients who need mitral valve replacement, and more research on the relations between AF and angiotensin converting enzyme would be needed. PMID- 18390219 TI - [Causes of rise of troponin after percutaneous coronary intervention and its clinical implication]. AB - OBJECTIVE: To analyse the clinical factors that contribute to the rise of the cardiac injury marker troponin after coronary intervention and the impact of the rise of troponin on the clinical outcomes. METHODS: Troponin I was measured after elective coronary intervention in 129 patients whose baseline levels of troponin were normal. The clinical outcomes of the patients were follow-up. RESULTS: The rise of troponin I was associated with side branch losses, flow impairments such as no flow or slow flow and diabetes. The incidence of myocardial infarction increased with the rise of troponin I. The angina onsets were more common and the length of stay were longer in the patients with the rise of troponin I than those without. CONCLUSION: The rise of troponin after coronary intervention is related to the complex coronary lesions and the complications of intervention procedures. To a certain extent, the level of troponin can predict the patients' outcomes. PMID- 18390220 TI - [Cytokines of splenocytes in mice by different vaccination with recombinant BCG Em14-3-3 vaccines against Echinococcus multilocularis]. AB - OBJECTIVE: To detect the changes of cytokines of splenocytes in mice immunized by recombinant BCG-Em14-3-3 vaccines against Echinococcus multilocularis (Em). METHODS: BALB/c mice were subcutaneously and intranasally vaccinated by the BCG Em14-3-3 vaccines respectively, with PBS as a control. The mice were challenged with Em protoscoleces eight weeks after the vaccinations. The spleens of the mice were collected 18 weeks after the infections. The splenocytes were separated and cultured with the stimulation of EmAg or ConA. The IL-2, IFN-gamma, TNF-alpha and IL-4 in the supernatants were measured with ELISA. RESULTS: Significant increase of IFN-gamma and TNF-alpha were found in the immunized mice. But the IL-4 was decreased after the vaccinations. The levels of TNF-alpha were higher in the intranasal vaccinated mice than in the subcutaneous vaccinated mice. CONCLUSIONS: TH1 response is induced in mice by rBCG-Em14-3-3 vaccines to fight against the challenge of Em protoscoleces. Intranasal vaccinations are preferable. PMID- 18390221 TI - [Detecting nicotine and its metabolite cotinine in human hair with capillary gas chromatography]. AB - OBJECTIVE: To develop a rapid method for determination of nicotine and its metabolite cotinine in human hair with capillary gas chromatography with flame ionization detector. METHODS: The hair sample was digested by 1.5 mol/L sodium hydroxide solution. The nicotine and cotinine in the hair sample were extracted with a mixed solvent of dichloromethane-methanol (3:1). Aliquot of the extraction solution was vaporized with nitrogen flow and then methanol was added to dissolve the analysts. The analysts were tested with capillary gas chromatography. RESULTS: The detection limits (signal to noise ratio of 3:1) were 4.3 ng/mL for nicotine and 10 ng/mL for cotinine, respectively. The spiked recoveries were 90.33%-113.1% for nicotine with relative standard deviation (RSD) of 2.1%-7.0% and 92.92%-117.4% for cotinine with relative standard deviation (RSD) of 4.4% 8.9%, respectively. CONCLUSION: The proposed method is sensitive, simple, easy and rapid. It can be easily extended to large scale applications in tobacco epidemiology studies. PMID- 18390223 TI - The practice of general surgery today requires growth and adaptability. PMID- 18390222 TI - [Determination and Weibull analysis of the strength of dental veneering porcelains]. AB - OBJECTIVE: To determine the strength of the veneering porcelain stored in artificial saliva. METHODS: Standard beams with a size of 3 mm x 2 mm x 20 mm made from the body, incisal, and translucent porcelains of SHOFU Vintage were stored in the artificial saliva for 30 days. Twenty samples from each material were tested for the strength by the 3-point flexural test at a crosshead speed of 0.5 mm. The data were analyzed by the two-parameter Weibull method. RESULTS: The strength, Weibull modulus (m) and characterization strength (male0) were (75.5 +/ 5.6) MPa, 17.3, and 77.6 MPa for Body porcelain, respectively; (73.6 +/- 5.0) MPa, 16.5, and 76.0 MPa for incisal porcelain, respectively; and (72.9 +/- 4.5) MPa, 16.1, and 75. 5 MPa for translucent porcelain, respectively. CONCLUSION: There are no differences in strength between the three dental porcelains. Weibull analysis describes the strength of the porcelains better than the average and standard deviation. PMID- 18390224 TI - Chartered value exchanges. PMID- 18390225 TI - The coming era of too few physicians. PMID- 18390226 TI - The impact of transparency on patient safety and liability. PMID- 18390227 TI - Risk management programs: a means to lower premiums. PMID- 18390228 TI - The value of teaching anatomy to medical students. PMID- 18390229 TI - A model for national health care. PMID- 18390230 TI - How to enhance interest in surgical specialties. PMID- 18390231 TI - Statistical match of the March 1996 Current Population Survey and the 1995 National Health Interview Survey. AB - OBJECTIVES: Statistical matching is a method used to combine two files when it is unlikely that individuals on one file are also on the other file. The objectives of this report are to document and evaluate statistical matches of the March 1996 Current Population Survey (CPS) and the 1995 National Health interview Survey (NHIS) and give recommendations for improving future matches. The CPS-NHIS match was motivated by the need for a data set with data on health measures and family resources for use in policy analyses. METHODS: Three statistical matches between the March 1996 CPS and the 1995 NHIS are described in this report. All three matches used person-level constrained matching with partitioning and a predictive mean matching algorithm to link records on the two files. For two of the matches, the CPS served as the Host file and the NHIS served as the Donor file; for the third match, the NHIS was the Host file and the CPS was the Donor file. RESULTS: The results suggest that the constrained predictive mean matches of the March 1996 CPS and the 1995 NHIS successfully combined some of the information on the two files, but that relationships among some Host and Donor variables on the matched file may be distorted. The evaluation of the matches suggested that the variables used to partition the Host and Donor files prior to matching and the variables involved in the predictive mean matching play an important role in determining whether relationships among variables on the matched file correctly represent relationships among those variables in the population. The evaluation also indicated that estimates for small subgroups may be especially subject to error. The results reinforce the need to proceed cautiously when exploring relationships among Host and Donor variables on a statistically matched file. PMID- 18390232 TI - The heart and soul... PMID- 18390233 TI - An update on the dermatoses of pregnancy. AB - CONTEXT: The dermatoses of pregnancy represent a group of pathologic skin disturbances that cause considerable distress to the gravid patient. It is important to understand the diagnosis, treatment, and implications of these disorders in order to avert unnecessary discomfort and complications. OBJECTIVES: To describe the specific dermatoses of pregnancy according to an established classification system. DATA SOURCES: Reports identified from PubMed and textbook searches using relevant terms pertaining to the stated questions. CONCLUSION: The dermatoses of pregnancy represent a unique group of disease processes caused or exacerbated by the gravid state. This includes gestational pemphigoid (GP), pruritic urticarial papules and plaques of pregnancy (PUPPP), prurigo of pregnancy (PP), intrahepatic cholestasis of pregnancy (ICP), and impetigo herpetiformis (IH). GP, ICP, and IH may result in serious complications, whereas PUPPP and PP are generally benign processes. Early recognition of these disorders will possibly reduce maternal and fetal morbidity and mortality. PMID- 18390234 TI - Do systemic corticosteroids improve acute outcomes in infants with RSV bronchiolitis? PMID- 18390235 TI - An update on breast cancer in Oklahoma and the dietary changes women make after diagnosis. AB - CONTEXT: The breast cancer (BC) death rate in Oklahoma exceeds that of the United States. Women commonly experience weight gain after diagnosis and soy isoflavones may interfere with effect of tamoxifen. OBJECTIVE: To assess dietary changes of BC survivors and to determine if the women desire information about diet from physicians. DESIGN, SETTING, AND PARTICIPANTS: Retrospective cross-sectional study of women BC survivors in clinic and community settings in central Oklahoma. MAIN OUTCOME MEASURES: Dietary changes, use of soy foods and supplements, physician advice about dietary changes. RESULTS: A total of 224 BC survivors were surveyed (81 in clinics and 143 at Susan G. Komen Race for the Cure). Median weight change was zero kg (IQR -0.23 to 4.55 kg). Mean weight change was 1.6 kg (95% CI -0.1 to 3.4 kg). Half the women said they had taken tamoxifen. Of those, 78% said they had never consumed or were eating the same amount of soy foods. Only 16% of women on tamoxifen ate more soy foods. Only 30% of women received any specific dietary advice from their physician, but over half said they would like more information. Most common dietary changes included decreased consumption of sugar, sweets, and regular soda, red meats, shortening, margarine, high fat dairy products, and alcohol. Women ate more olive oil, whole grains, vegetables, fruit, poultry, and low-fat dairy foods. PMID- 18390236 TI - A finite element analysis of two different dental implants: stress distribution in the prosthesis, abutment, implant, and supporting bone. AB - This study evaluates the influence of 2 commercially available dental implant systems on stress distribution in the prosthesis, abutment, implant, and supporting alveolar bone under simulated occlusal forces, employing a finite element analysis. The implants and abutments evaluated consisted of a stepped cylinder implant connected to a screw-retained, internal, hexagonal abutment (system 1) and a conical implant connected to a solid, internal, conical abutment (system 2). A porcelain-covered, silver-palladium alloy was used as a crown. In each case, a simulated, 100-N vertical load was applied to the buccal cusp. A finite element model was created based on the physical properties of each component, and the values of the von Mises stresses generated in the prosthesis, abutment, implant, and supporting alveolar bone were calculated. In the prostheses, the maximum von Mises stresses were concentrated at the points of load application in both systems, and they were greater in system 1 (148 N/mm2) than in system 2 (55 N/mm2). Stress was greater on the abutment of system 2 than of system 1 on both the buccal (342 N/mm2 x 294 N/mm2) and lingual (294 N/mm2 x 148 N/ mm2) faces. Stress in the cortical, alveolar bone crest was greater in system 1 than in system 2 (buccal: 99.5 N/mm2 x 55 N/mm2, lingual: 55 N/mm2 x 24.5 N/mm2, respectively). Within the limits of this investigation, the stepped cylinder implant connected to a screw-retained, internal hexagonal abutment produces greater stresses on the alveolar bone and prosthesis and lower stresses on the abutment complex. In contrast, the conical implant connected to a solid, internal, conical abutment furnishes lower stresses on the alveolar bone and prosthesis and greater stresses on the abutment. PMID- 18390237 TI - Measurement of the fatigue life of mini dental implants: a pilot study. AB - The fatigue life of mini or small-diameter dental implants is of particular interest because these implants are used to retain and support fixed and removable dental prostheses. The fatigue life of an implant depends on both the implant itself as well as on the physical properties of the bone. However, the capability to predict the fatigue life of a newly placed implant is currently inexistent. This pilot study represents the first step in developing such a methodology and focuses on the design of a cost-effective device to measure the fatigue life of a dental implant. In our measurements, the implant has been mounted in an essentially rigid support, but test specimens can also be bone mounted in vitro. Furthermore, we developed a finite element-based computer model capable of predicting the corresponding fatigue life. The finite element analysis was performed in ABAQUS, and the results predicted by the model correlated fairly well with our initial experimental results. Most of the 2-mm diameter implants fractured after more than a million cycles. PMID- 18390238 TI - Histologic analysis of human peri-implant bone in type 1 osteoporosis. AB - Osteoporosis is a disease that influences the quality of bone tissue. At present, osteoporosis represents a contraindication or a risk factor for osseointegration. The aim of this report was to evaluate the bone-to-implant contact of 2 loaded implants retrieved after prosthetic failure in a woman with type 1 osteoporosis. Histologically, only one implant was osseointegrated, and it appeared surrounded by healthy bone tissue. The bone-to-implant contact presented a mean of 51.25%. No foreign body reaction was found at the bone-to-implant contact, although epithelial downgrowth was observed at the interface. Data from this case report demonstrate that the peri-implant bone histology of the dental implant retrieved from an osteoporotic patient presented no alteration. However, the role of osteoporosis in the long-term success of dental implants needs further investigation. PMID- 18390239 TI - Bone response to modified titanium surface implants in nonhuman primates (Papio ursinus) and humans: histological evaluation. AB - The aim of this study is a comparative histological and histomorphometrical evaluation of the effect on early bone formation of 2 different implant surfaces: a machined and a new acid-etched implant surface (Leader, Milano, Italy). Ten screw-type microimplants were placed in 5 patients. Each patient received 2 microimplants (2 mm in diameter and 5 mm in length): 1 with a machined surface (control) and 1 with an acid-etched surface (test). The microimplants were retrieved after 60 days of healing with a 4-mm trephine bur and processed for histology. Moreover, 24 regular size implants--12 with a machined surface (control) and 12 with an acid-etched surface (test)--were placed in 2 adult nonhuman primates 3 months after the extraction of premolars and molars. Each animal received 3 machined implants (control) in the right hemimandible and 3 acid-etched implants (test) in the left hemimandible. The same animals received 3 control implants and 3 test implants in the rectus abdominis muscle. After 1 month, the implants were retrieved from the mandible and the rectus abdominis muscle and processed for histology. Histomorphometric evaluation demonstrated a higher bone-to-implant contact in the test implants compared with the controls in both primates (25.55% vs 15.8%) and humans (62% vs 45%). Moreover, in nonhuman primates after 1 month of healing, it was possible to observe a poor osseointegration in the control specimens, while newly formed bone in direct contact with test implants was evident. The rectus abdominis muscle specimens showed that the acid-etched surfaces can stimulate the formation and attachment of new connective and vascular tissues more than machined surfaces can. Implant surface geometry can speed up bone formation by the development of a special microenvironment that promotes angiogenesis. Long-term studies are needed to further test this new acid-etched implant surface. PMID- 18390240 TI - Analysis of a rapid, simple, and inexpensive technique used to obtain platelet rich plasma for use in clinical practice. AB - The use of platelet-rich plasma (PRP) has become more generally accepted, and implant dentists are using PRP more frequently to promote the healing of oral surgical and/or periodontal wounds. Critical elements of PRP are thought to be growth factors contained within the concentrated platelets. These growth factors are known to promote soft-tissue healing, angiogenesis and osteogenesis. We present a rapid, simple, and inexpensive methodology for preparing PRP using the Cliniseal centrifuge method. This study demonstrates that platelets are concentrated approximately 6-fold without altering platelet morphology. Further we demonstrate that key growth factors, platelet-derived growth factor BB (PDGF BB), transforming growth factor B (TGF-B1), vasculature endothelial growth factor (VEGF), and epidermal growth factor (EGF) are present in comparable or higher concentrations than those reported with the use of other techniques. Prolonged bench set time (>3 hours) after centrifugation resulted in decreased concentration of TGF-B1 but not decreased concentration of PDGF-BB, VEGF, or EGF. This study confirms the molecular aspects of PRP obtained using this inexpensive and efficient methodology. PMID- 18390241 TI - Avoiding injury to the inferior alveolar nerve by routine use of intraoperative radiographs during implant placement. AB - Injury to the inferior alveolar nerve during implant placement in the posterior atrophic mandible is a rare but serious complication. Although a preoperative computerized tomography scan can help determine the distance from the alveolar ridge to the nerve canal, variables such as magnification errors, ridge anatomy, and operator technique can increase the chance for complications. The routine use of intraoperative periapical radiographs during the drilling sequence is an inexpensive and reliable tool, allowing the operator to confidently adjust the direction and depth of the implant during placement. Most important, it helps avoid the risk of injury to the inferior alveolar nerve in cases in which there is limited vertical alveolar bone. Using this technique for 21 implants placed in the posterior atrophic mandible, with less than 10 mm of vertical bone to the inferior alveolar nerve canal, the authors observed no incidents of postoperative paresthesia. PMID- 18390242 TI - Five-year prospective clinical evaluation of highly crystalline HA MP-1-coated dental implants. AB - Contemporary plasma-sprayed hydroxylapatite (HA) coatings with high crystalline content are much more resistant to in vivo degradation than HA coatings of a decade ago but reportedly exhibit reduced wettability, which could potentially negatively affect tissue adhesion and long-term clinical outcome. The present prospective study was undertaken to determine if highly crystalline HA MP-1 coated implants could meet a minimum 5-year implant success rate standard of 85% in view of their previously reported decreased wettability. Study subjects were consecutive patients with 1 or more missing teeth in the maxillary and/or mandibular jaw who presented in 3 university dental clinics and 1 private dental practice. A total of 120 patients were treated per protocol and successfully restored with implant-supported prostheses. Four implants failed in 3 patients and were withdrawn from the study. There were no other irresolvable adverse events. Cumulative implant success at 5 years was 97% (n = 184 implants in 88 patients), which exceeded the 85% standard for implant success after 5 years of clinical function. PMID- 18390243 TI - An alternative method to treat a case with severe maxillary atrophy by the use of angled implants instead of complicated augmentation procedures: a case report. AB - Several treatment options with implants have been described for maxillary edentulous patients. Maxillary implant-supported overdentures have been shown to be a predictable, accepted treatment option for the edentulous maxilla. Patients with severe bone resorption present additional difficulties, and implant treatment in the atrophic maxilla represents a challenge. Anatomical limitations and patient desires in this case have forced the treatment to be 4 angulated implants supporting an upper overdenture. Since conventional single-retention mechanisms such as ball (O-ring), locator, or telescopes would transfer too much force to the implants, especially because of their angulation, an individual bar was fabricated. One-year follow-up of the case showed a stable peri-implant condition on bone as well as soft tissue level. Although further follow-up and higher case numbers will give more information about this treatment modality, the actual result is encouraging and can be recommended for similar cases. PMID- 18390244 TI - Immediate flapless implant placement and provisionalization: challenge for optimum esthetics and function: a case report. AB - This report presents a case of tooth extraction and immediate flapless implant placement followed by fabrication of transitional restoration. The tooth was extracted atraumatically with the use of a periotome followed by careful debridement. An osteotomy was performed up to 5 mm beyond the base of the socket and depth using the alveolar crest as a landmark, following a slightly palatal direction. The implant shoulder was inserted 3 mm below the cementoenamel junction of the adjacent tooth. The interproximal distance from the neighboring teeth was 3 mm. No membranes and/or grafts were used. Initial impressions were taken immediately after implant placement; 6 hours later a well-polished and slightly overcontoured (at the distal-mesial aspect) acrylic crown was fixed onto the implant. There were no contacts in the centric and lateral positions. Five months later, the occlusion was modified allowing slight contacts in the centric position for an additional 2 months. The final prosthetic restoration was placed 2 months later (7 months after surgery), consisting of a full ceramic crown cemented on a customized metal ceramic UCLA abutment. The technique maintained the integrity of hard and soft tissues and created a very favorable esthetic result. It also provided the patient with a transitional fixed restoration and reduced the time required for therapy completion. Because research on this field is limited, further investigation is required to support the results of this report, despite the promising clinical outcome. PMID- 18390245 TI - Fabrication of a retrievable cement- and screw-retained implant-supported zirconium fixed partial denture: a case report. AB - It can be difficult to achieve superior esthetics in implant-supported fixed partial dentures (FPDs). Zirconia ceramics with high flexural strength and esthetic can be treatment options for implant-supported FPDs. This article describes a simple and reliable method to fabricate a retrievable cemented implant-supported zirconia FPD. PMID- 18390246 TI - On the Lucentis controversy. PMID- 18390247 TI - Data briefing. Why budgets have always been a bugbear. PMID- 18390249 TI - Arches, peacocks and horses. PMID- 18390248 TI - Log in to the future. PMID- 18390250 TI - A national dental practice survey. AB - The very high correlations from the dentist surveys reinforce the contention that the practice of dentistry is similar across this country without regard for geographic region). The dental school responses provide validation for the perceptions of the dentists surveyed regarding the important content in dental practice. The complete practice analysis report is available at http:// www.wreb.org. PMID- 18390251 TI - Incidence of use of unregulated herbal products by patients in sampled Texas dental practices. PMID- 18390252 TI - Oral and maxillofacial pathology case of the month. Intraosseous schwannoma. PMID- 18390253 TI - If you were a patient in pain, wouldn't it be great if every month were pain awareness month? PMID- 18390254 TI - Differences in blood pressure values obtained with automated and manual methods in medical inpatients. AB - Measurement of blood pressure is done frequently in hospitalized patients with an automated, noninvasive blood pressure device. While manufacturers have made significant design improvements which may have improved the accuracy of these devices, limited research is available on their performance in clinical practice. The purpose of this study was to determine the accuracy of a contemporary automated, noninvasive blood pressure device in hospitalized medical patients. PMID- 18390255 TI - A review of the use of the health belief model for weight management. AB - Research found in the professional literature on the Health Belief Model and its use in weight management programs is reviewed. Within this model, the primary motivation to change is the level of perceived threat or risk of a specific condition (obesity), and the primary resource for change is self-efficacy or confidence to make the change to lose or maintain a healthy weight. PMID- 18390256 TI - Recognizing excellence: unit-based activities to support specialty nursing certification. AB - An acute care medicine unit identified and implemented strategies to encourage staff to become certified. In 1 year, the number of certified nurses increased 60%, the RN turnover rate decreased from 16.7% to 8.1%, the RN vacancy rate decreased from 11% to 4.73%, and the patient satisfaction score increased. The processes that contributed to those changes are described. PMID- 18390257 TI - Effectiveness of an intervention for disseminating Cochrane reviews to nurses. AB - Disseminating research findings to clinicians can be a challenging task. In this study, researchers tested the effectiveness of disseminating summaries of systematic reviews to clinicians. The findings suggest that these summaries are well-accepted by nurses and increase their knowledge of research findings. PMID- 18390258 TI - A day with Blake: hope on a medical-surgical unit. AB - All nurses care for patients who change their lives forever. Blake was one of those patients. Blake's story and the interventions his nurse (Amie) instituted illustrate a hope-promoting framework that medical-surgical nurses can use every day in practice. PMID- 18390259 TI - Self-care agency, health promoting lifestyle, and satisfaction with life in postmenopausal women who mall walk. AB - Walking as exercise is a self-care, health-promoting activity of many older adults. In this article, self-care agency, health-promoting lifestyle, and satisfaction with life in two groups of postmenopausal women--those who mall walk and those who do not engage in regular exercise--are compared. PMID- 18390260 TI - Deciphering the diagnostics of breast cancer. AB - Today's medical-surgical nurse working in the hospital, clinic, or community needs to understand the diverse diagnostic screening tools used to screen and evaluate breast cancer in the United States. Risk assessment models, mammography, ultrasound, breast MRI, genetic testing, and cancer prevention are discussed. PMID- 18390261 TI - Patient safety: the ethical imperative. PMID- 18390262 TI - Nurse practitioners and physician assistants: do you know the difference? PMID- 18390263 TI - Nursing students benefit your facility. PMID- 18390264 TI - ECG of the month. Electrocardiogram in a woman with rheumatic heart disease. Normal sinus rhythm, left atrial enlargement with intra-atrial block; left ventricular hypertrophy with repolarization abnormality. PMID- 18390265 TI - An immunosuppressed woman presenting with acute flaccid paralysis and progressive respiratory failure. PMID- 18390266 TI - Radiology case of the month. Neck pain after motor vehicle accident. Cervical acceleration-deceleration injury (Whiplash) resulting in muscle sprain best seen in MR examination. PMID- 18390267 TI - Hemangioendothelioma. PMID- 18390268 TI - Penetrating orbitocranial injury to the sella: case report and review of the literature. AB - A 39-year-old woman sustained a self-inflicted transorbital penetrating injury that resulted in direct pituitary injury with hypopituitarism and decreased vision in the opposite eye. Several hormone deficiencies were detected. Even though this patient did not develop any other complications from her injury (abscess, cerebrospinal fluid fistula or pseudoaneurysm), after two years of follow-up no recovery of pituitary or visual function has occurred. CONCLUSION: From our experience in this case, we propose a conservative multidisciplinary approach when dealing with this type of lesion. Broad spectrum antibiotic coverage and early detection and replacement of any hormone deficiency should be instituted. PMID- 18390269 TI - Creutzfeldt-Jakob disease in Louisiana, 1980-2006. PMID- 18390270 TI - Pancreas transplantation--a controversy in evolution. AB - Pancreas transplantation has been mired in controversy throughout its existence. Arguments have erupted regarding its actual indications, the way the surgical procedure should be performed, its benefits, and today, the concept of pancreas islet cell transplantation remains controversial as well. If diabetic patients had a choice between life long insulin therapy and a major operation with immunosuppression afterward, what would they choose? The answer may not be as easy as one thinks. Pancreas transplantation has come a long way. This manuscript discusses the history of pancreas transplantation, how the indications are starting to be defined, evolution of the surgical procedure, current success rates of this procedure, the current scenario of pancreas islet transplantation, and newer developing technologies. PMID- 18390271 TI - Acute necrotizing esophagitis: case report and review of the literature. AB - Acute necrotizing esophagitis (ANE) is a rare disease typically presenting as a diffuse blackening of the esophageal mucosa with necrosis on histologic examination. For this reason, ANE is often considered to be synonymous with'black esophagus'. Recently, however, cases of ANE have been reported with a "non-black" esophagus, and this is believed to be an earlier or later stage in the evolution of ANE to/from its black form. Here we present a case report of ANE that illustrates such an evolution in endoscopic appearance from non-black to black esophagus. We also provide a review of the literature to highlight what is currently known about this color change and other clinical aspects of this relatively uncommon esophageal disease. PMID- 18390272 TI - Time for change. PMID- 18390273 TI - [Feline paraneoplastic alopecia]. AB - A 14-year-old neutered female cat was presented for a second opinion regarding hair loss, anorexia, and lethargy. Examination revealed alopecia of the ventral chest, abdomen, axilla, and groin. The footpads were encrusted. The liver was enlarged and nodular. Investigation revealed an enlarged liver with multiple nodules, and small and atrophic hair follicles. Paraneoplastic alopecia due to hepatocellular carcinoma was diagnosed and subsequently confirmed postmortem. PMID- 18390275 TI - [Long teeth]. PMID- 18390274 TI - [Chemical and biological dangers to a horse veterinarian who becomes exposed]. PMID- 18390276 TI - [Horse Advisory Committee III--Infectious complications--digestion (part II)]. PMID- 18390277 TI - [Change in veterinary drug regulation]. PMID- 18390278 TI - [Parafilariasis, are we seeing it again in 2008?]. PMID- 18390279 TI - [Biochemical cause of muscular disease discovered]. PMID- 18390280 TI - [Highlights DSK]. PMID- 18390282 TI - How to evaluate tracking systems. An instrument of efficiency. PMID- 18390281 TI - Resolving to make patient safety a priority. Interview by Alan Joch. PMID- 18390283 TI - Seeing is believing: materials stance on price transparency. PMID- 18390284 TI - Communication between finance and materials counts. PMID- 18390285 TI - Up in arms over clean hands. AB - Hand hygiene has been proven to be one of the most effective, cost-efficient methods of controlling nosocomial infections. For that reason, the hand hygiene practices of health care workers are being scrutinized more than ever. This article focuses on some of the many questions health care workers have about hand hygiene practices. PMID- 18390286 TI - GS1 Healthcare US helps push data standards for industry. PMID- 18390287 TI - Smart pumps need OK from clinicians. PMID- 18390288 TI - Monitoring products within perioperative services is vital. PMID- 18390289 TI - Neighborhood selection and the social reproduction of concentrated racial inequality. AB - In this paper, we consider neighborhood selection as a social process central to the reproduction of racial inequality in neighborhood attainment. We formulate a multilevel model that decomposes multiple sources of stability and change in longitudinal trajectories of achieved neighborhood income among nearly 4000 Chicago families followed for up to seven years wherever they moved in the United States. Even after we adjust for a comprehensive set of fixed and time-varying covariates, racial inequality in neighborhood attainment is replicated by movers and stayers alike. We also study the emergent consequences of mobility pathways for neighborhood-level structure. The temporal sorting by individuals of different racial and ethnic groups combines to yield a structural pattern offlows between neighborhoods that generates virtually nonoverlapping income distributions and little exchange between minority and white areas. Selection and racially shaped hierarchies are thus mutually constituted and account for an apparent equilibrium of neighborhood inequality. PMID- 18390290 TI - Maternal work hours and adolescents' school outcomes among low-income families in four urban counties. AB - We examine how changes in maternal work hours affect adolescent children's school participation and performance outcomes using data from interviews in 1998 and 2001 with approximately 1700 women who, in May 1995, were welfare-reliant, single mothers of adolescents living in neighborhoods of concentrated poverty in Cuyahoga (Cleveland), Los Angeles, Miami-Dade, and Philadelphia counties. Analyses control for a broad array of mothers' characteristics, including their psychological and physical health, experiences with domestic violence and substance abuse, as well as unobserved time-invariant characteristics. In fixed effects models, we find unfavorable effects of increased maternal work hours on three of six outcomes: skipping school, performing above average, and parental contact about behavior problems. Adolescent-aged sons seem to be particularly sensitive to changes in mothers' hours of work. PMID- 18390291 TI - The neighborhood context of racial and ethnic disparities in arrest. AB - This study assesses the role of social context in explaining racial and ethnic disparities in arrest, with afocus on how distinct neighborhood contexts in which different racial and ethnic groups reside explain variations in criminal outcomes. To do so, I utilize a multilevel, longitudinal research design, combining individual-level data with contextual data from the Project on Human Development in Chicago Neighborhoods (PHDCN). Findings reveal that black youths face multiple layers of disadvantage relative to other racial and ethnic groups, and these layers work to create differences in arrest. At the family level, results show that disadvantages in the form of unstable family structures explain much of the disparities in arrest across race and ethnicity. At the neighborhood level, black youths tend to reside in areas with both significantly higher levels of concentrated poverty than other youths as well as lower levels of collective efficacy than white youths. Variations in neighborhood tolerance of deviance across groups explain little of the arrest disparities, yet tolerance of deviance does influence the frequency with which a crime ultimately ends in an arrest. Even after accounting for relevant demographic, family, and neighborhood-level predictors, substantial residual arrest differences remain between black youths and youths of other racial and ethnic groups. PMID- 18390292 TI - Immigrant residential segregation in U.S. metropolitan areas, 1990-2000. AB - This paper examines the extent of spatial assimilation among immigrants of different racial and ethnic origins. We use restricted data from the 1990 and 2000 censuses to calculate the levels of dissimilarity by race and Hispanic origin, nativity, and year of entry, and then run multivariate models to examine these relationships. The findings provide broad support for spatial assimilation theory. Foreign-born Hispanics, Asians, and blacks are more segregated from native-born non-Hispanic whites than are the U.S.-born of these groups. The patterns for Hispanics and Asians can be explained by the average characteristics of the foreign-born that are generally associated with higher levels of segregation, such as lower levels of income, English language ability, and home ownership. We also find that immigrants who have been in the United States for longer periods are generally less segregated than new arrivals, and once again, much of this difference can be attributed to the characteristics of immigrants. However, patterns also vary across groups. Levels of segregation are much higher for black immigrants than for Asian, Hispanic, and white immigrants. In addition, because black immigrants are, on average, of higher socioeconomic status than native-born blacks, such characteristics do not help explain their very high levels of segregation. PMID- 18390293 TI - Using subjective expectations to forecast longevity: do survey respondents know something we don't know? AB - Old-age mortality is notoriously difficult to predict because it requires not only an understanding of the process of senescence-which is influenced by genetic, environmental, and behavioral factors-but also a prediction of how these factors will evolve. In this paper I argue that individuals are uniquely qualified to predict their own mortality based on their own genetic background, as well as environmental and behavioral risk factors that are often known only to the individual. Given this private information, individuals form expectations about survival probabilities that may provide additional information to demographers and policymakers in their challenge to predict mortality. From expectations data from the 1992 Health and Retirement Study (HRS), I construct subjective, cohort life tables that are shown to predict the unusual direction of revisions to U.S. life expectancy by gender between 1992 and 2004: that is, for these cohorts, the Social Security Actuary (SSA) raised male life expectancy in 2004 and at the same lowered female life expectancy, narrowing the gender gap in longevity by 25% over this period. Further, although the subjective life expectancies for men appear to be roughly in line with the 2004 life tables, the subjective expectations of women suggest that female life expectancies estimated by the SSA might still be on the high side. PMID- 18390294 TI - Retrospective information on health status and its application for population health measures. AB - Healthy life expectancies are almost always calculated by using health data from cross-sectional surveys. This type of calculation is done partly because data from longitudinal surveys are not always available, and when they are available, they are collected at intervals that are longer than one year. In such cases, collecting health information retrospectively for the years skipped by the survey is useful. The main purpose of this paper is to show how retrospective health information can be used to estimate life expectancies in different health states. Healthy life expectancies are estimated with and without using data on retrospective health information, and the corresponding estimates are compared. The two sets of estimates are similar. We conclude that retrospectively assessed health information based on a one-year recall period can be used to estimate years of life in various health states and that estimates based on such information will closely approximate estimates based on concurrent health information. PMID- 18390295 TI - The quality of retrospective data on cohabitation. AB - We assess the quality of retrospective data on cohabitation by comparing data collected in four major U.S. family surveys: the National Survey of Families and Households and three rounds of the National Survey of Family Growth. We use event history analysis to analyze rates of entry into cohabitation in age-period-cohort segments captured by multiple surveys. We find consistent discrepancies among the four surveys. The pattern of differences suggests that cohabitation histories underestimate cohabitation rates in distant periods relative to rates estimated closer to the date of survey. We conclude with cautions regarding the use of retrospective data on cohabitation. PMID- 18390296 TI - A practical approach to using multiple-race response data: a bridging method for public-use microdata. AB - Revised federal policies require that multiple-race responses be allowed in all federal data collection efforts, but many researchers find the multitude of race categories and variables very difficult to use. Important comparability issues also interfere with using multiple-race data in analyses of multiple data sets and/or several points in time. These difficulties have, in effect, discouraged the use of the new data on race. We present a practical method for incorporating multiple-race respondents into analyses that use public-use microdata. Our method is a modification of the regression method developed by the National Center for Health Statistics (NCHS), which uses multiple-race respondents' specific combination of races, as well as other individual-level and contextual characteristics, to predict the respondents 'preferred single race. In this paper we (1) apply the NCHS-generated regression coefficients to public-use microdata with limited geographic information; and (2) provide a downloadable computer program with which researchers can apply this practical and preferable method for including multiple-race respondents in a wide variety of analyses. PMID- 18390297 TI - The multistate life table method: an application to contraceptive switching behavior. AB - In many demographic behaviors (e.g., those relating to marriage, contraception, migration, and health), people change among multiple statuses through time, sometimes leaving and then returning to the same status. Data on such behaviors are often collected in surveys as censored event histories. The multistate life table (MSLT) can be used to properly describe, in a single analysis, these complex transitions among multiple states measured in such data, but MSLT is rarely applied in the demographic literature because practical guidance is lacking on how to compute MSLTs with such data. We provide methods for computing MSLT quantities using censored event-history data: namely, transition intensities and probabilities, "state occupancy" probabilities and standard errors, average time spent in specified states, and average number of visits to specified states. Applying these methods to contraceptive use, we find high levels of switching back and forth, particularly between barrier methods and non-use, resulting in high rates of unintended pregnancy. PMID- 18390298 TI - The international child poverty gap: does demography matter? AB - According to the Luxembourg Income Study data, the U.S. child poverty rate is the second highest among 15 high-income nations. The present work reveals that 55% of all American children living in a household headed by a single female with no other adult present live in poverty-the highest rate for any of the five living arrangements in the 15 countries examined in this study. While previous analyses have focused on market forces and governmental redistribution across households, we question the contribution of demographic factors that place children in family structures with different poverty risks relative to other factors such as differential market opportunities and governmental benefits for adults caring for children in various living arrangements. Applying a classic demographic decomposition technique to the overall poverty gap, we find that the distributional effect of demographic behavior contributes little to the U.S. poverty gap with other nations (and none with respect to the United Kingdom). Overall differences in labor markets and welfare schemes best explain the U.S. child poverty gap, although for some countries, the gap is accentuated by the gradient of governmental transfers, and for most countries, by the gradient of market earnings across living arrangements. PMID- 18390299 TI - Cohort estimates of nonmarital fertility for U.S. women. AB - Historical trends in U.S. nonmarital fertility have been compiled almost exclusively from vital statistics on births. This paper complements this historical record by providing cohort estimates of nonmarital fertility for cohorts of U.S. women spanning approximately 50 years of cohort experience. Life table estimates using retrospective marital and fertility histories in the June 1980, 1985, 1990, and 1995 Current Population Surveys reveal nonnegligible levels of nonmarital fertility historically. For women born between 1925 and 1929, nearly 1 in 10 had at least one nonmarital birth by age 30. For women born between 1965 and 1969, more than 1 of 4 had one or more nonmarital births by age 30, with roughly 1 of5 white, 3 of 5 black, and 1 in 3 Hispanic women having at least one nonmarital birth by age 30. Life table estimates reveal a twofold increase between ages 20 and 30 in the percentage of women with at least one child outside of formal marriage for all cohorts of white and Hispanic women, and an increase of roughly two-thirds for all cohorts of black women. I also document qualitative differences in nonmarital fertility by race/ethnicity, with the percentage of nonmarital births following a divorce or marital separation for white women approximately twice that for black or Hispanic women. Finally, I introduce a new measure, the cohort nonmarital fertility ratio (CNMFR), which provides a cohort complement to the standard period nonmarital fertility ratio. Conservative estimates reveal a roughly threefold increase in the CNMFR for women born from 1925-1929 to 1950-1954 for both whites and blacks, despite substantially higher levels of nonmarital fertility among black women. Overall, these findings reveal surprisingly high levels of nonmarital fertility for women born since the 1920s and confirm that nonmarital fertility has become an increasingly substantial component of overall U.S. fertility. PMID- 18390300 TI - Moving and union dissolution. AB - This paper examines the effect of migration and residential mobility on union dissolution among married and cohabiting couples. Moving is a stressful life event, and a large, multidisciplinary literature has shown that family migration often benefits one partner (usually the man) more than the other Even so, no study to date has examined the possible impact of within-nation geographical mobility on union dissolution. We base our longitudinal analysis on retrospective event-history data from Austria. Our results show that couples who move frequently have a significantly higher risk of union dissolution, and we suggest a variety of mechanisms that may explain this. PMID- 18390301 TI - The quantity-quality trade-off of children in a developing country: identification using Chinese twins. AB - Testing the trade-off between child quantity and quality within a family is complicated by the endogeneity of family size. Using data from the Chinese Population Census, we examine the effect of family size on child educational attainment in China. We find a negative correlation between family size and child outcome, even after we control for the birth order effect. We then instrument family size by the exogenous variation that is induced by a twin birth and find a negative effect offamily size on children's education. We also find that the effect of family size is more evident in rural China, where the public education system is poor. Given that our estimates of the effect of having twins on nontwins at least provide the lower bound of the true effect of family size, these findings suggest a quantity-quality trade-off for children in developing countries. PMID- 18390303 TI - Delay-dependent criteria for global robust periodicity of uncertain switched recurrent neural networks with time-varying delay. AB - In this paper, we introduce some ideas of switched systems into the field of neural networks and a large class of switched recurrent neural networks (SRNNs) with time-varying structured uncertainties and time-varying delay is investigated. Some delay-dependent robust periodicity criteria guaranteeing the existence, uniqueness, and global asymptotic stability of periodic solution for all admissible parametric uncertainties are devised by taking the relationship between the terms in the Leibniz-Newton formula into account. Because free weighting matrices are used to express this relationship and the appropriate ones are selected by means of linear matrix inequalities (LMIs), the criteria are less conservative than existing ones reported in the literature for delayed neural networks with parameter uncertainties. Some examples are given to show that the proposed criteria are effective and are an improvement over previous ones. PMID- 18390302 TI - A Franco-British patients' and partners' satisfaction audit of breast reconstruction. AB - AIMS: To compare French and British patients' and partners' satisfaction with breast reconstruction after mastectomy and determine what factors affect it. METHODOLOGY: The study analysed validated questionnaires from 182 pairs (the woman and her partner), split equally between France and the UK. The patients underwent breast surgery at the Norfolk and Norwich University Hospital (NNUH) and at the Hopital Saint-Louis in Paris, France. All main surgical procedures were considered. RESULTS: Participants were satisfied with the outcome but satisfaction levels differed between France and the UK, British patients reporting higher satisfaction (P=0.029) because of a closer match between their expectation and the outcome. There were also statistically significant differences in: There was no difference in satisfaction levels between patients and partners who accurately reported on patients' views and whose own views matched patients'. Satisfaction was correlated to the level of information, pain management, recovery time, technique used, cosmetic results, intervention timing, but not to the independent assessment of the outcome. PMID- 18390304 TI - A one-layer recurrent neural network with a discontinuous hard-limiting activation function for quadratic programming. AB - In this paper, a one-layer recurrent neural network with a discontinuous hard limiting activation function is proposed for quadratic programming. This neural network is capable of solving a large class of quadratic programming problems. The state variables of the neural network are proven to be globally stable and the output variables are proven to be convergent to optimal solutions as long as the objective function is strictly convex on a set defined by the equality constraints. In addition, a sequential quadratic programming approach based on the proposed recurrent neural network is developed for general nonlinear programming. Simulation results on numerical examples and support vector machine (SVM) learning show the effectiveness and performance of the neural network. PMID- 18390305 TI - Locality-preserved maximum information projection. AB - Dimensionality reduction is usually involved in the domains of artificial intelligence and machine learning. Linear projection of features is of particular interest for dimensionality reduction since it is simple to calculate and analytically analyze. In this paper, we propose an essentially linear projection technique, called locality-preserved maximum information projection (LPMIP), to identify the underlying manifold structure of a data set. LPMIP considers both the within-locality and the between-locality in the processing of manifold learning. Equivalently, the goal of LPMIP is to preserve the local structure while maximize the out-of-locality (global) information of the samples simultaneously. Different from principal component analysis (PCA) that aims to preserve the global information and locality-preserving projections (LPPs) that is in favor of preserving the local structure of the data set, LPMIP seeks a tradeoff between the global and local structures, which is adjusted by a parameter alpha, so as to find a subspace that detects the intrinsic manifold structure for classification tasks. Computationally, by constructing the adjacency matrix, LPMIP is formulated as an eigenvalue problem. LPMIP yields orthogonal basis functions, and completely avoids the singularity problem as it exists in LPP. Further, we develop an efficient and stable LPMIP/QR algorithm for implementing LPMIP, especially, on high-dimensional data set. Theoretical analysis shows that conventional linear projection methods such as (weighted) PCA, maximum margin criterion (MMC), linear discriminant analysis (LDA), and LPP could be derived from the LPMIP framework by setting different graph models and constraints. Extensive experiments on face, digit, and facial expression recognition show the effectiveness of the proposed LPMIP method. PMID- 18390306 TI - Shared feature extraction for nearest neighbor face recognition. AB - In this paper, we propose a new supervised linear feature extraction technique for multiclass classification problems that is specially suited to the nearest neighbor classifier (NN). The problem of finding the optimal linear projection matrix is defined as a classification problem and the Adaboost algorithm is used to compute it in an iterative way. This strategy allows the introduction of a multitask learning (MTL) criterion in the method and results in a solution that makes no assumptions about the data distribution and that is specially appropriated to solve the small sample size problem. The performance of the method is illustrated by an application to the face recognition problem. The experiments show that the representation obtained following the multitask approach improves the classic feature extraction algorithms when using the NN classifier, especially when we have a few examples from each class. PMID- 18390307 TI - Complex ICA by negentropy maximization. AB - In this paper, we use complex analytic functions to achieve independent component analysis (ICA) by maximization of non-Gaussianity and introduce the complex maximization of non-Gaussianity (CMN) algorithm. We derive both a gradient descent and a quasi-Newton algorithm that use the full second-order statistics providing superior performance with circular and noncircular sources as compared to existing methods. We show the connection among ICA methods through maximization of non-Gaussianity, mutual information, and maximum likelihood (ML) for the complex case, and emphasize the importance of density matching for all three cases. Local stability conditions are derived for the CMN cost function that explicitly show the effects of noncircularity on convergence and demonstrated through simulation examples. PMID- 18390308 TI - Large-scale maximum margin discriminant analysis using core vector machines. AB - Large-margin methods, such as support vector machines (SVMs), have been very successful in classification problems. Recently, maximum margin discriminant analysis (MMDA) was proposed that extends the large-margin idea to feature extraction. It often outperforms traditional methods such as kernel principal component analysis (KPCA) and kernel Fisher discriminant analysis (KFD). However, as in the SVM, its time complexity is cubic in the number of training points m, and is thus computationally inefficient on massive data sets. In this paper, we propose an (1+epsilon)(2)-approximation algorithm for obtaining the MMDA features by extending the core vector machine. The resultant time complexity is only linear in m, while its space complexity is independent of m. Extensive comparisons with the original MMDA, KPCA, and KFD on a number of large data sets show that the proposed feature extractor can improve classification accuracy, and is also faster than these kernel-based methods by over an order of magnitude. PMID- 18390309 TI - DCT-Yager FNN: a novel Yager-based fuzzy neural network with the discrete clustering technique. AB - Earlier clustering techniques such as the modified learning vector quantization (MLVQ) and the fuzzy Kohonen partitioning (FKP) techniques have focused on the derivation of a certain set of parameters so as to define the fuzzy sets in terms of an algebraic function. The fuzzy membership functions thus generated are uniform, normal, and convex. Since any irregular training data is clustered into uniform fuzzy sets (Gaussian, triangular, or trapezoidal), the clustering may not be exact and some amount of information may be lost. In this paper, two clustering techniques using a Kohonen-like self-organizing neural network architecture, namely, the unsupervised discrete clustering technique (UDCT) and the supervised discrete clustering technique (SDCT), are proposed. The UDCT and SDCT algorithms reduce this data loss by introducing nonuniform, normal fuzzy sets that are not necessarily convex. The training data range is divided into discrete points at equal intervals, and the membership value corresponding to each discrete point is generated. Hence, the fuzzy sets obtained contain pairs of values, each pair corresponding to a discrete point and its membership grade. Thus, it can be argued that fuzzy membership functions generated using this kind of a discrete methodology provide a more accurate representation of the actual input data. This fact has been demonstrated by comparing the membership functions generated by the UDCT and SDCT algorithms against those generated by the MLVQ, FKP, and pseudofuzzy Kohonen partitioning (PFKP) algorithms. In addition to these clustering techniques, a novel pattern classifying network called the Yager fuzzy neural network (FNN) is proposed in this paper. This network corresponds completely to the Yager inference rule and exhibits remarkable generalization abilities. A modified version of the pseudo-outer product (POP)-Yager FNN called the modified Yager FNN is introduced that eliminates the drawbacks of the earlier network and yi- elds superior performance. Extensive experiments have been conducted to test the effectiveness of these two networks, using various clustering algorithms. It follows that the SDCT and UDCT clustering algorithms are particularly suited to networks based on the Yager inference rule. PMID- 18390310 TI - Real-time reconfigurable subthreshold CMOS perceptron. AB - In this paper, a new, real-time reconfigurable perceptron circuit element is presented. A six-transistor version used as a threshold gate, having a fan-in of three, producing adequate outputs for threshold of T =1, 2 and 3 is demonstrated by chip measurements. Subthreshold operation for supply voltages in the range of 100-350 mV is shown. The circuit performs competitively with a standard static complimentary metal-oxide-semiconductor (CMOS) implementation when maximum speed and energy delay product are taken into account, when used in a ring oscillator. Functionality per transistor is, to our knowledge, the highest reported for a variety of comparable circuits not based on floating gate techniques. Statistical simulations predict probabilities for making working circuits under mismatch and process variations. The simulations, in 120-nm CMOS, also support discussions regarding lower limits to supply voltage and redundancy. A brief discussion on how the circuit may be exploited as a basic building block for future defect tolerant mixed signal circuits, as well as neural networks, exploiting redundancy, is included. PMID- 18390311 TI - Relevance-based feature extraction for hyperspectral images. AB - Hyperspectral imagery affords researchers all discriminating details needed for fine delineation of many material classes. This delineation is essential for scientific research ranging from geologic to environmental impact studies. In a data mining scenario, one cannot blindly discard information because it can destroy discovery potential. In a supervised classification scenario, however, the preselection of classes presents one with an opportunity to extract a reduced set of meaningful features without degrading classification performance. Given the complex correlations found in hyperspectral data and the potentially large number of classes, meaningful feature extraction is a difficult task. We turn to the recent neural paradigm of generalized relevance learning vector quantization (GRLVQ) [B. Hammer and T. Villmann, Neural Networks vol. 15, pp. 1059-1068, 2002], which is based on, and substantially extends, learning vector quantization (LVQ) [T. Kohonen, Self-Organizing Maps, Berlin, Germany: Springer-Verlag, 2001] by learning relevant input dimensions while incorporating classification accuracy in the cost function. By addressing deficiencies in GRLVQ, we produce an improved version, GRLVQI, which is an effective analysis tool for high-dimensional data such as remotely sensed hyperspectral data. With an independent classifier, we show that the spectral features deemed relevant by our improved GRLVQI result in a better classification for a predefined set of surface materials than using all available spectral channels. PMID- 18390312 TI - Output feedback stabilization for time-delay nonlinear interconnected systems using neural networks. AB - In this paper, dynamic output feedback control problem is investigated for a class of nonlinear interconnected systems with time delays. Decentralized observer independent of the time delays is first designed. Then, we employ the bounds information of uncertain interconnections to construct the decentralized output feedback controller via backstepping design method. Based on Lyapunov stability theory, we show that the designed controller can render the closed-loop system asymptotically stable with the help of the changing supplying function idea. Furthermore, the corresponding decentralized control problem is considered under the case that the bounds of uncertain interconnections are not precisely known. By employing the neural network approximation theory, we construct the neural network output feedback controller with corresponding adaptive law. The resulting closed-loop system is stable in the sense of semiglobal boundedness. The observers and controllers constructed in this paper are independent of the time delays. Finally, simulations are done to verify the effectiveness of the theoretic results obtained. PMID- 18390313 TI - PSECMAC: a novel self-organizing multiresolution associative memory architecture. AB - The cerebellum constitutes a vital part of the human brain system that possesses the capability to model highly nonlinear physical dynamics. The cerebellar model articulation controller (CMAC) associative memory network is a computational model inspired by the neurophysiological properties of the cerebellum, and it has been widely used for control, optimization, and various pattern recognition tasks. However, the CMAC network's highly regularized computing structure often leads to the following: 1) a suboptimal modeling accuracy, 2) poor memory utilization, and 3) the generalization-accuracy dilemma. Previous attempts to address these shortcomings have limited success and the proposed solutions often introduce a high operational complexity to the CMAC network. This paper presents a novel neurophysiologically inspired associative memory architecture named pseudo-self-evolving CMAC (PSECMAC) that nonuniformly allocates its computing cells to overcome the architectural deficiencies encountered by the CMAC network. The nonuniform memory allocation scheme employed by the proposed PSECMAC network is inspired by the cerebellar experience-driven synaptic plasticity phenomenon observed in the cerebellum, where significantly higher densities of synaptic connections are located in the frequently accessed regions. In the PSECMAC network, this biological synaptic plasticity phenomenon is emulated by employing a data-driven adaptive memory quantization scheme that defines its computing structure. A neighborhood-based activation process is subsequently implemented to facilitate the learning and computation of the PSECMAC structure. The training stability of the PSECMAC network is theoretically assured by the proof of its learning convergence, which will be presented in this paper. The performance of the proposed network is subsequently benchmarked against the CMAC network and several representative CMAC variants on three real-life applications, namely, pricing of currency futures option, banking failure classification, and modeling of the glucose-insulin dynamics of the human glucose metabolic process. The experimental results have strongly demonstrated the effectiveness of the PSECMAC network in addressing the architectural deficiencies of the CMAC network by achieving significant improvements in the memory utilization, output accuracy as well as the generalization capability of the network. PMID- 18390314 TI - Adaptive importance sampling to accelerate training of a neural probabilistic language model. AB - Previous work on statistical language modeling has shown that it is possible to train a feedforward neural network to approximate probabilities over sequences of words, resulting in significant error reduction when compared to standard baseline models based on n-grams. However, training the neural network model with the maximum-likelihood criterion requires computations proportional to the number of words in the vocabulary. In this paper, we introduce adaptive importance sampling as a way to accelerate training of the model. The idea is to use an adaptive n-gram model to track the conditional distributions produced by the neural network. We show that a very significant speedup can be obtained on standard problems. PMID- 18390315 TI - A Note on the bias in SVMs for multiclassification. AB - During the usual SVM biclassification learning process, the bias is chosen a posteriori as the value halfway between separating hyperplanes. A note on different approaches on the calculation of the bias when SVM is used for multiclassification is provided and empirical experimentation is carried out which shows that the accuracy rate can be improved by using bias formulations, although no single formulation stands out as providing better performance. PMID- 18390316 TI - Further results on delay-dependent stability criteria of neural networks with time-varying delays. AB - In this brief paper, an augmented Lyapunov functional, which takes an integral term of state vector into account, is introduced. Owing to the functional, an improved delay-dependent asymptotic stability criterion for delayed neural networks (NNs) is derived in term of linear matrix inequalities (LMIs). It is shown that the obtained criterion can provide less conservative result than some existing ones. When linear fractional uncertainties appear in NNs, a new robust delay-dependent stability condition is also given. Numerical examples are given to demonstrate the applicability of the proposed approach. PMID- 18390317 TI - Simulation of patient specific cervical hip fracture surgery with a volume haptic interface. AB - The interest for surgery simulator systems with anatomical models generated from authentic patient data is growing as these systems evolve. With access to volumetric patient data, e.g., from a computer tomography scan, haptic and visual feedback can be created directly from this dataset. This opens the door for patient specific simulations. Hip fracture surgery is one area where simulator systems is useful to train new surgeons and plan operations. To simulate the drilling procedure in this type of surgery, a repositioning of the fractured bone into correct position is first needed. This requires a segmentation process in which the bone segments are identified and the position of the dislocated part is determined. The segmentation must be automatic to cope with the large amount of data from the computer tomography scan. This work presents the first steps in the development of a hip fracture surgery simulation with patient specific models. Visual and haptic feedback is generated from the computer tomography data by simulating fluoroscopic images and the drilling process. We also present an automatic segmentation method to identify the fractured bone and determine the dislocation. This segmentation method is based on nonrigid registration with the Morphon method. PMID- 18390318 TI - A mathematical model of respiratory and biothermal dynamics in brain hypothermia treatment. AB - Brain hypothermia treatment (BHT) requires proper mechanical ventilation and therapeutic cooling. The cooling strategy for BHT has been mainly discussed in the literature while little information is available on the respiratory management. We first developed a mathematical model that integrates the respiratory and biothermal dynamics to discuss the simultaneous managements of mechanical ventilation and therapeutic cooling. The effect of temperature on the linear approximations of hemoglobin-oxygen dissociation, together with temperature dependency of metabolism, is introduced during modeling to combine the respiratory system with the biothermal system. By comparing its transient behavior with published data, the model is verified qualitatively and then quantitatively. Second, model-based simulation of the current respiratory management in BHT suggests reduction of minute ventilation in reference to cooled brain temperature to stabilize the states of blood and brain oxygenation. Lastly, the relationship between cooling temperature and minute ventilation is approximated by a linear first-order transfer function of static gain 0.61min(-1) degrees C(-1) and time constant 8.9 h, which is used to develop a feedforward control to tune the mechanical ventilator in concert with temperature regulation of the cooling blanket. Discussion of the model encourages further studies that provide direct evidence from clinical experiments. PMID- 18390319 TI - Numerical design of experiment for sensitivity analysis--application to skin burn injury prediction. AB - Temperature evolution and skin burn process resulting from a laser radiation exposure are investigated in this paper. Transient temperature in skin is numerically estimated using a 1-D multilayered model based on Penne's equation. The degree of burn injury is numerically evaluated by using an Arrhenius-type function. Unfortunately, most of the mathematical model parameters are not well defined in literature. Thus, a sensitivity analysis has been performed in order to evaluate the effect of each parameters inaccuracy on temperature estimation and on burn injuries prediction (according to several authors' characterization). Investigated parameters uncertainties that crucially invalidate the thermal model are as follows: epidermis and dermis volumetric heat, extinction coefficient, and skin thickness of the affected area. Considering the damage prediction, the activation energy is a key parameter for the validation of an efficient predictive tool. PMID- 18390320 TI - Relations between the timing of the second heart sound and aortic blood pressure. AB - The second heart sound (S2) is triggered by an aortic valve closure as a result of the ventricular-arterial interaction of the cardiovascular system. The objective of this paper is to investigate the timing of S2 in response to the changes in hemodynamic parameters and its relation to aortic blood pressure (BP). An improved model of the left ventricular-arterial interaction was proposed based on the combination of the newly established pressure source model of the ventricle and the nonlinear pressure-dependent compliance model of the arterial system. The time delay from the onset of left ventricular pressure rise to the onset of S2 (RS2) was used to measure the timing of S2. The results revealed that RS2 bears a strong negative correlation with both systolic blood pressure and diastolic blood pressure under the effect of changing peripheral resistance, heart rate, and contractility. The results were further validated by a series of measurements of 16 normal subjects submitted to dynamic exercise. This study helps understand the relationship between the timing of S2 and aortic BP under various physiological and pathological conditions. PMID- 18390321 TI - Modeling cellular metabolism and energetics in skeletal muscle: large-scale parameter estimation and sensitivity analysis. AB - Skeletal muscle plays a major role in the regulation of whole-body energy metabolism during physiological stresses such as ischemia, hypoxia, and exercise. Current experimental techniques provide relatively little in vivo data on dynamic responses of metabolite concentrations and metabolic fluxes in skeletal muscle to such physiological stimuli. As a complementary approach to experimental measurements and as a framework for quantitatively analyzing available in vivo data, a physiologically based model of skeletal muscle cellular metabolism and energetics is developed. This model, which incorporates key transport and reaction processes, is based on dynamic mass balances of 30 chemical species in capillary (blood) and tissue (cell) domains. The reaction fluxes in the cellular domain are expressed in terms of a generalized Michaelis?Menten equation involving energy controller ratios ATP/ADP and ATP/ADP and NADH/NAD+ . This formalism introduces a large number of unknown parameters ( approximately 90). Estimating these parameters from in vivo sparse data and evaluating dynamic sensitivities of the model outputs with respect to these parameters is a challenging problem. Parameter estimation is accomplished using an efficient, nonlinear, constraint-based, optimization algorithm that minimizes differences between available experimental data and corresponding model outputs by explicitly utilizing equality constraints on resting fluxes and concentrations. With the estimated parameter values, the model is able to simulate dynamic responses to reduced blood flow (ischemia) of key metabolite concentrations and metabolic fluxes, both measured and nonmeasured. A general parameter sensitivity analysis is carried out to determine and characterize the parameters having the most and least effects on the measured outputs. PMID- 18390322 TI - An ECG signals compression method and its validation using NNs. AB - This paper presents a new algorithm for electrocardiogram (ECG) signal compression based on local extreme extraction, adaptive hysteretic filtering and Lempel-Ziv-Welch (LZW) coding. The algorithm has been verified using eight of the most frequent normal and pathological types of cardiac beats and an multi-layer perceptron (MLP) neural network trained with original cardiac patterns and tested with reconstructed ones. Aspects regarding the possibility of using the principal component analysis (PCA) to cardiac pattern classification have been investigated as well. A new compression measure called "quality score," which takes into account both the reconstruction errors and the compression ratio, is proposed. PMID- 18390323 TI - Truncated total least squares: a new regularization method for the solution of ECG inverse problems. AB - The reconstruction of epicardial potentials (EPs) from body surface potentials (BSPs) can be characterized as an ill-posed inverse problem which generally requires a regularized numerical solution. Two kinds of errors/noise: geometric errors and measurement errors exist in the ECG inverse problem and make the solution of such problem more difficulty. In particular, geometric errors will directly affect the calculation of transfer matrix A in the linear system equation AX = B. In this paper, we have applied the truncated total least squares (TTLS) method to reconstruct EPs from BSPs. This method accounts for the noise/errors on both sides of the system equation and treats geometric errors in a new fashion. The algorithm is tested using a realistically shaped heart-lung torso model with inhomogeneous conductivities. The h-adaptive boundary element method [h-BEM, a BEM mesh adaptation scheme which starts from preset meshes and then refines (adds/removes) grid with fixed order of interpolation function and prescribed numerical accuracy] is used for the forward modeling and the TTLS is applied for inverse solutions and its performance is also compared with conventional regularization approaches such as Tikhonov and truncated single value decomposition (TSVD) with zeroth-, first-, and second-order. The simulation results demonstrate that TTLS can obtain similar results in the situation of measurement noise only but performs better than Tikhonov and TSVD methods where geometric errors are involved, and that the zeroth-order regularization is the optimal choice for the ECG inverse problem. This investigation suggests that TTLS is able to robustly reconstruct EPs from BSPs and is a promising alternative method for the solution of ECG inverse problems. PMID- 18390324 TI - Respiratory sounds compression. AB - Recently, with the advances in digital signal processing, compression of biomedical signals has received great attention for telemedicine applications. In this paper, an adaptive transform coding-based method for compression of respiratory and swallowing sounds is proposed. Using special characteristics of respiratory sounds, the recorded signals are divided into stationary and nonstationary portions, and two different bit allocation methods (BAMs) are designed for each portion. The method was applied to the data of 12 subjects and its performance in terms of overall signal-to-noise ratio (SNR) values was calculated at different bit rates. The performance of different quantizers was also considered and the sensitivity of the quantizers to initial conditions has been alleviated. In addition, the fuzzy clustering method was examined for classifying the signal into different numbers of clusters and investigating the performance of the adaptive BAM with increasing the number of classes. Furthermore, the effects of assigning different numbers of bits for encoding stationary and nonstationary portions of the signal were studied. The adaptive BAM with variable number of bits was found to improve the SNR values of the fixed BAM by 5 dB. Last, the possibility of removing the training part for finding the parameters of adaptive BAMs for each individual was investigated. The results indicate that it is possible to use a predefined set of BAMs for all subjects and remove the training part completely. Moreover, the method is fast enough to be implemented for real-time application. PMID- 18390325 TI - Robust unsupervised detection of action potentials with probabilistic models. AB - We develop a robust and fully unsupervised algorithm for the detection of action potentials from extracellularly recorded data. Using the continuous wavelet transform allied to probabilistic mixture models and Bayesian probability theory, the detection of action potentials is posed as a model selection problem. Our technique provides a robust performance over a wide range of simulated conditions, and compares favorably to selected supervised and unsupervised detection techniques. PMID- 18390326 TI - Analysis and measurements of magnetic field exposures for healthcare workers in selected MR environments. AB - There are concerns about workers repeatedly exposed to magnetic fields exceeding regulatory limits with respect to modern magnetic resonance imaging (MRI). As a result, there is need for an ambulatory magnetic field dosimeter capable of measuring these fields in and around an MRI scanner in order to evaluate the regulatory guidelines and determine any underlying exposure risks. This study presents results of tri-axial measurements using an ambulatory magnetic field dosimeter worn by workers during normal working shifts. We recorded and analyzed magnetic field exposures in and around 1.5 T, 2 T, and 4 T magnets during routine patient procedures. The data was integrated and averaged over time and evaluated against the latest exposure standards. Time-varying magnetic fields occur when individuals move through spatially non-uniform static magnetic fields or during gradient-pulsed magnetic fields or a combination of both. Our previous numerical analysis shows that at certain positions surrounding the MRI scanner ends, such fields may induce current densities and electric fields that may exceed the relevant EU, ICNIRP, and IEEE standards. A high-speed acquisition version of the dosimeter measured gradient- pulsed fields at positions accessible by MRI workers near the scanner ends, and the results were evaluated and compared against the numerical simulations and the standards. Our measurements confirm that workers can be exposed to magnetic fields exceeding the guidelines at positions near the gradient coil ends during clinical imaging and a high degree of correlation exists with the numerical results. While the time-weighted average magnetic field exposures in 1.5 T, 2 T, and 4 T were all within the regulatory limits during static magnetic field measurements, the peak limits for the head can be exceeded in some circumstances. This study presents a small number of routine shifts of data that provide indicative results of magnetic field exposure in real situations. PMID- 18390327 TI - Designing the optical interface of a transcutaneous optical telemetry link. AB - Optical telemetry has long been an option for transcutaneous data transfer and has been used in various types of implanted systems. This telemetry modality and the efficiency of these optical links are becoming ever more important as higher bandwidth sources such as cortical recording arrays are being implemented in implanted systems. The design of the transmitter-skin-receiver interface (the "optical interface") is paramount to the operation of a transcutaneous optical telemetry link. This interface functions to achieve sufficient receiver signal power for data communication. This paper describes a mathematical analysis and supporting data that quantitatively describes the relationship between the primary interface design parameters. These parameters include the thickness of the skin through which the light is transmitted, the size of the integration area of the optics, the degree of transmitter-receiver misalignment, the efficiency of the optics system, and the emitter power. The particular combination of these parameters chosen for the hardware device will determine the receiver signal power and, therefore, the data quality for the link. This paper demonstrates some of the tradeoffs involved in the selection of these design parameters and provides suggestions for link design. This analysis may also be useful for transcutaneous optical powering systems. PMID- 18390328 TI - A telemetric pressure sensor system for biomedical applications. AB - A new implantable pressure sensor for long-term monitoring of intracranial pressure is presented. The sensor is powered by telemetry and can be interrogated wirelessly. A capacitive pressure transducer, whose capacitance is converted to a frequency-encoded signal by an application-specific integrated circuit (ASIC), senses the absolute pressure. The pressure-encoded signal, the ASIC input voltage, and onboard calibration parameters are transmitted to an external reading unit. The proposed novel packaging solution is designed for long-term stability and reliability of the sensor. The accuracy of sensor at body temperature is better than 2 mbar across a pressure range of 600-1200 mbar. The sensor is 13 mm in diameter and 4.5 mm in height. PMID- 18390329 TI - Analysis of cultured neuronal networks using intraburst firing characteristics. AB - It is an open question whether neuronal networks, cultured on multielectrode arrays, retain any capability to usefully process information (learning and memory). A necessary prerequisite for learning is that stimulation can induce lasting changes in the network. To observe these changes, one needs a method to describe the network in sufficient detail, while stable in normal circumstances. We analyzed the spontaneous bursting activity that is encountered in dissociated cultures of rat neocortical cells. Burst profiles (BPs) were made by estimating the instantaneous array-wide firing frequency. The shape of the BPs was found to be stable on a time scale of hours. Spatiotemporal detail is provided by analyzing the instantaneous firing frequency per electrode. The resulting phase profiles (PPs) were estimated by aligning BPs to their peak spiking rate over a period of 15 min. The PPs reveal a stable spatiotemporal pattern of activity during bursts over a period of several hours, making them useful for plasticity and learning studies. We also show that PPs can be used to estimate conditional firing probabilities. Doing so, yields an approach in which network bursting behavior and functional connectivity can be studied. PMID- 18390330 TI - Simulation studies of ultrashort, high-intensity electric pulse induced action potential block in whole-animal nerves. AB - A theoretical study of possible neuromuscular incapacitation based on the application of high-intensity, ultrashort electric pulses is presented. The analysis is applied to a rat, but the approach is general and can be extended to any whole-animal and applies for any arbitrary pulse waveform. It is hypothesized that repeatable and reversible action potential blocks in nerves can be attained based on the electroporation mechanism. Our numerical studies are based on the Hodgkin-Huxley distributed circuit representation of nerves, and incorporate a nodal analysis for the time-dependent and volumetric perturbing potentials and internal electric fields in whole animals. The predictions are compared to actual 600-ns experimental reports on rats and shown to be in very good agreement. Effective strength-duration plots for neuromuscular incapacitation are also generated. PMID- 18390331 TI - Optimal filter design to compute the mean of cardiovascular pressure signals. AB - The mean pressure is a term used to describe the baseline trend of physiological pressure signals that excludes fluctuations due to the cardiac cycle and, in some cases, the respiratory cycle. In many clinical applications and bedside monitoring devices, the mean pressure is estimated with a 3-8 s moving average. We suggest that the mean pressure is best defined in terms of its frequency domain properties. This definition makes it possible to determine solutions that are both optimal and practical. We demonstrate that established methods of optimal finite impulse response (FIR) filter design produce estimates of the mean pressure that are significantly more accurate than the moving average. These filters have no more computational cost, are less sensitive to artifact, have shorter delays, and greater sensitivity to acute events. PMID- 18390332 TI - Linear electrode arrays for stimulation and recording within cardiac tissue space constants. AB - In this paper, we document a fabrication process that yields linear arrays of rectangular platinum black electrodes spaced 25 mum apart with edge-to-edge separation of 20 microm. The spatial arrangement is therefore sufficiently fine to insure stimulation and recording within cardiac tissue space constants, as six electrodes with dimensions of either 5 x 100 microm2, 5 x 250 microm2, or 5 x 500 microm2 were positioned in a 130-microm2 span in the arrays. Despite the small electrode sizes and available surface areas, favorable impedance characteristics were identifed. Averages ranged from 111 kOmega to 146 kOmega at 0.5 Hz and from 14 kOmega 39 kOmega at 500 Hz. Differences in impedances between the electrode sizes tested were small. Potential differences (deltaphis) recorded using the two central electrodes during stimulation with combinations at separations of only 75 microm, 100 microm, and 125 microm had low signal noise. As a preliminary test of the use of these arrays for possible application to impedance measurements in cardiac tissue, the deltaphis recorded during stimulation were compared to deltaphis obtained from finite-difference simulations using an isotropic volume conductor model. Anticipated decays in deltaphi with widening electrode separation identified in those simulations matched the decays in the recorded deltaphis closely. These findings are significant because they suggest intracellular and interstitial microimpedance mesurements in heart experiments will be straightforward. PMID- 18390333 TI - New accelerometric method to discriminate between asymptomatic subjects and patients with medial knee osteoarthritis during 3-d gait. AB - This study presents a new method to estimate 3-D linear accelerations at tibial and femoral functional coordinate systems. The method combines the use of 3-D accelerometers, 3-D gyroscopes and reflective markers rigidly fixed on an exoskeleton and, a functional postural calibration method. Marker positions were tracked by a six-camera optoelectronic system (VICON 460, Oxford Metrics). The purpose of this study was to determine if this method could discriminate between medial osteoarthritic and asymptomatic knees during gait. Nine patients with osteoarthritic knees and nine asymptomatic control subjects were included in this study. Eighteen parameters representing maximal, minimal, and range of acceleration values were extracted during the loading and preswing to mid-swing phase periods, and were compared in both groups. Results show good discriminative capacity of the new method. Eight parameters were significantly different between both groups. The proposed method has the potential to be used in comprehending and monitoring gait strategy in patients with osteoarthritic knee. PMID- 18390334 TI - The effect of resultant force at the pushrim on shoulder kinetics during manual wheelchair propulsion: a simulation study. AB - The aim of this study was to determine, by simulation on real data, the effect of modifying the direction or effectiveness of a given force amplitude on the load sustained by the shoulder estimated by joint forces and moments. Kinematics and kinetics data were recorded on 14 manual wheelchair users 68.2+/-5.2 years for 10 s at sub-maximal speed (0.96-1.01 m/s). The simulation consisted in modifying force effectiveness at the pushrim while maintaining the same initial force amplitude. Shoulder kinetics were computed for simulated resultant forces from radial to tangent directions and also for initial force effectiveness. The results show that as the force was simulated tangent to the wheel, there was a significant increase in the average proximal and anterior shoulder joint forces. Also, significant increases in average internal rotation, flexion in the sagittal and horizontal plane moments were reported. Higher shoulder kinetics could accelerate the onset of fatigue and increase the risk of injury. A single-case analysis revealed an improvement window for force effectiveness ( approximately 10%) in which shoulder kinetics were not substantially increased. Our results provide useful information on what would happen to shoulder kinetics if we were able to teach manual wheelchair users to modify their force pattern at the pushrim. The results suggest that for an elderly population, it is not wise to aim at producing a mechanically optimal resultant force at the pushrim (i.e., tangent). Smaller increases of the initial force effectiveness would be preferable. PMID- 18390335 TI - Feasibility of sonophoretic delivery for effective skin optical clearing. AB - In an attempt to find a noninvasive way to improve the efficacy of skin optical clearing with topical application of optical clearing agents (OCA), we exploited the effect of ultrasound on optical clearing of intact skin in vitro Changes in optical transmittance and diffuse reflectance of the porcine skin under different treatment procedures were measured by a UV/Vis/NIR spectrophotometer. Clearing capability of glycerol or propylene glycol was much improved with the simultaneous application of ultrasound with a frequency of 1 MHz and a power of 0.75 W over a 3-cm probe. Light transmittance at 470 and 1276 nm increased by 112%-128% and 54%-65%, respectively. The most significant effect was seen where skin was treated with the combination of ultrasound and the chemical enhancer azone, resulting in a 2.3-fold increase of optical clearing at 1276 nm. We demonstrate the feasibility of using sonophoretic delivery methods to improve skin optical clearing with topical application of OCA. PMID- 18390336 TI - Anatomical optical coherence tomography for long-term, portable, quantitative endoscopy. AB - In this paper, we report on anatomical optical coherence tomography, a catheter based optical modality designed to provide quantitative sectional images of internal hollow organ anatomy over extended observational periods. We consider the design and performance of an instrument and its initial intended application in the human upper airway for the characterization of obstructive sleep apnea (OSA). Compared with current modalities, the technique uniquely combines quantitative imaging, bedside operation, and safety for use over extended periods of time with no cumulative dose limit. Our experiments show that the instrument is capable of imaging subjects during sleep, and that it can record dynamic changes in airway size and shape. PMID- 18390337 TI - The influence of porous silicon on axonal outgrowth in vitro. AB - xonal outgrowth on smooth and porous silicon surfaces was studied in organ culture. The pore size of the silicon substrata varied between 100 and 1500 nm. We found that axons preferred to grow and elongate on porous silicon surfaces only when pores of (150-500 nm) are available. PMID- 18390338 TI - Investigation of short-term changes in visual evoked potentials with windowed adaptive chirplet transform. AB - We propose a new application of the adaptive chirplet transform that involves partitioning signals into non-overlapping sequential segments. From these segments, the local time-frequency structures of the signal are estimated by using a four-parameter chirplet decomposition. Entitled the windowed adaptive chirplet transform (windowed ACT), this approach is applied to the analysis of visual evoked potentials (VEPs). It can provide a unified and compact representation of VEPs from the transient buildup to the steady-state portion with less computational cost than its non-windowed counterpart. This paper also details a method to select the optimal window length for signal segmentation. This approach will be useful for long-term signal monitoring as well as for signal feature extraction and data compression. PMID- 18390339 TI - The influence of CSF on EEG sensitivity distributions of multilayered head models. AB - We examined how the cerebrospinal fluid (CSF) affects the distribution of electroencephalogram (EEG) measurement sensitivity. We used concentric spheres and realistic head models to investigate the difference between computed tomography (CT) and magnetic resonance image (MRI) models that exclude the CSF layer. The cortical EEG sensitivity distributions support these phenomena and show that the CSF layer significantly influences them, thus identifying the importance of including the CSF layer inside the head volume conductor models. The results show that the highly conductive CSF channels the current, thus decreasing the maximum cortical current density relative to models that do not include the CSF. We found that the MRI and CT models yielded HSV results 20% and 45%, respectively, too small when compared with CSF-inclusive models. PMID- 18390340 TI - The dependence of spectral impedance on disc microelectrode radius. AB - As microelectrodes gain widespread use for electrochemical sensing, biopotential recording, and neural stimulation, it becomes important to understand the dependence of electrochemical impedance on microelectrode size. It has been shown mathematically that a disc electrode, coplanar in an insulating substrate and exposed to a conducting media, exhibits an inhomogeneous current distribution when a potential step is applied. This distribution is known as the primary distribution, and its derivation also yielded an analytic solution for electrical resistance of the conducting media (R(s)), between the disc surface and a distant ground, which is inversely proportional to disk radius [R(s) = 1/(4kappar), where kappa is media conductivity and r is disk radius]. The dependence of spectral impedance on microelectrode radius, however, has not been explored. We verify the analytical solution for resistance using high-frequency (100 kHz) electrochemical impedance data from microelectrodes of varying radius (11-325 microm). For all disc radii, as we approach a lower frequency (--> 10 Hz), we observe a transition from radial to area dependence (e.g., 1/r --> 1/r2). We hypothesize that this transition is driven by the fact that the derivation of the primary distribution ignores concentration gradients, but that these gradients cannot be ignored at lower frequencies. PMID- 18390341 TI - An optimized blockwise nonlocal means denoising filter for 3-D magnetic resonance images. AB - A critical issue in image restoration is the problem of noise removal while keeping the integrity of relevant image information. Denoising is a crucial step to increase image quality and to improve the performance of all the tasks needed for quantitative imaging analysis. The method proposed in this paper is based on a 3-D optimized blockwise version of the nonlocal (NL)-means filter (Buades, et al., 2005). The NL-means filter uses the redundancy of information in the image under study to remove the noise. The performance of the NL-means filter has been already demonstrated for 2-D images, but reducing the computational burden is a critical aspect to extend the method to 3-D images. To overcome this problem, we propose improvements to reduce the computational complexity. These different improvements allow to drastically divide the computational time while preserving the performances of the NL-means filter. A fully automated and optimized version of the NL-means filter is then presented. Our contributions to the NL-means filter are: 1) an automatic tuning of the smoothing parameter; 2) a selection of the most relevant voxels; 3) a blockwise implementation; and 4) a parallelized computation. Quantitative validation was carried out on synthetic datasets generated with BrainWeb (Collins, et al., 1998). The results show that our optimized NL-means filter outperforms the classical implementation of the NL means filter, as well as two other classical denoising methods [anisotropic diffusion (Perona and Malik, 1990)] and total variation minimization process (Rudin, et al., 1992) in terms of accuracy (measured by the peak signal-to-noise ratio) with low computation time. Finally, qualitative results on real data are presented . PMID- 18390343 TI - Quantification of thyroid volume using 3-D ultrasound imaging. AB - Ultrasound (US) is among the most popular diagnostic techniques today. It is non invasive, fast, comparably cheap, and does not require ionizing radiation. US is commonly used to examine the size, and structure of the thyroid gland. In clinical routine, thyroid imaging is usually performed by means of 2-D US. Conventional approaches for measuring the volume of the thyroid gland or its nodules may therefore be inaccurate due to the lack of 3-D information. This work reports a semi-automatic segmentation approach for the classification, and analysis of the thyroid gland based on 3-D US data. The images are scanned in 3 D, pre-processed, and segmented. Several pre-processing methods, and an extension of a commonly used geodesic active contour level set formulation are discussed in detail. The results obtained by this approach are compared to manual interactive segmentations by a medical expert in five representative patients. Our work proposes a novel framework for the volumetric quantification of thyroid gland lobes, which may also be expanded to other parenchymatous organs. PMID- 18390342 TI - Fluid registration of diffusion tensor images using information theory. AB - We apply an information-theoretic cost metric, the symmetrized Kullback-Leibler (sKL) divergence, or J-divergence, to fluid registration of diffusion tensor images. The difference between diffusion tensors is quantified based on the sKL divergence of their associated probability density functions (PDFs). Three dimensional DTI data from 34 subjects were fluidly registered to an optimized target image. To allow large image deformations but preserve image topology, we regularized the flow with a large-deformation diffeomorphic mapping based on the kinematics of a Navier-Stokes fluid. A driving force was developed to minimize the J-divergence between the deforming source and target diffusion functions, while reorienting the flowing tensors to preserve fiber topography. In initial experiments, we showed that the sKL-divergence based on full diffusion PDFs is adaptable to higher-order diffusion models, such as high angular resolution diffusion imaging (HARDI). The sKL-divergence was sensitive to subtle differences between two diffusivity profiles, showing promise for nonlinear registration applications and multisubject statistical analysis of HARDI data. PMID- 18390344 TI - Segmentation of pulmonary nodules in thoracic CT scans: a region growing approach. AB - This paper presents an efficient algorithm for segmenting different types of pulmonary nodules including high and low contrast nodules, nodules with vasculature attachment, and nodules in the close vicinity of the lung wall or diaphragm. The algorithm performs an adaptive sphericity oriented contrast region growing on the fuzzy connectivity map of the object of interest. This region growing is operated within a volumetric mask which is created by first applying a local adaptive segmentation algorithm that identifies foreground and background regions within a certain window size. The foreground objects are then filled to remove any holes, and a spatial connectivity map is generated to create a 3-D mask. The mask is then enlarged to contain the background while excluding unwanted foreground regions. Apart from generating a confined search volume, the mask is also used to estimate the parameters for the subsequent region growing, as well as for repositioning the seed point in order to ensure reproducibility. The method was run on 815 pulmonary nodules. By using randomly placed seed points, the approach was shown to be fully reproducible. As for acceptability, the segmentation results were visually inspected by a qualified radiologist to search for any gross miss-segmentation. 84% of the first results of the segmentation were accepted by the radiologist while for the remaining 16% nodules, alternative segmentation solutions that were provided by the method were selected. PMID- 18390345 TI - Segmenting lung fields in serial chest radiographs using both population-based and patient-specific shape statistics. AB - This paper presents a new deformable model using both population-based and patient-specific shape statistics to segment lung fields from serial chest radiographs. There are two novelties in the proposed deformable model. First, a modified scale invariant feature transform (SIFT) local descriptor, which is more distinctive than the general intensity and gradient features, is used to characterize the image features in the vicinity of each pixel. Second, the deformable contour is constrained by both population-based and patient-specific shape statistics, and it yields more robust and accurate segmentation of lung fields for serial chest radiographs. In particular, for segmenting the initial time-point images, the population-based shape statistics is used to constrain the deformable contour; as more subsequent images of the same patient are acquired, the patient-specific shape statistics online collected from the previous segmentation results gradually takes more roles. Thus, this patient-specific shape statistics is updated each time when a new segmentation result is obtained, and it is further used to refine the segmentation results of all the available time-point images. Experimental results show that the proposed method is more robust and accurate than other active shape models in segmenting the lung fields from serial chest radiographs. PMID- 18390346 TI - Brain anatomical structure segmentation by hybrid discriminative/generative models. AB - In this paper, a hybrid discriminative/generative model for brain anatomical structure segmentation is proposed. The learning aspect of the approach is emphasized. In the discriminative appearance models, various cues such as intensity and curvatures are combined to locally capture the complex appearances of different anatomical structures. A probabilistic boosting tree (PBT) framework is adopted to learn multiclass discriminative models that combine hundreds of features across different scales. On the generative model side, both global and local shape models are used to capture the shape information about each anatomical structure. The parameters to combine the discriminative appearance and generative shape models are also automatically learned. Thus, low-level and high level information is learned and integrated in a hybrid model. Segmentations are obtained by minimizing an energy function associated with the proposed hybrid model. Finally, a grid-face structure is designed to explicitly represent the 3-D region topology. This representation handles an arbitrary number of regions and facilitates fast surface evolution. Our system was trained and tested on a set of 3-D magnetic resonance imaging (MRI) volumes and the results obtained are encouraging. PMID- 18390347 TI - MRI-based automated computer classification of probable AD versus normal controls. AB - Automated computer classification (ACC) techniques are needed to facilitate physician's diagnosis of complex diseases in individual patients. We provide an example of ACC using computational techniques within the context of cross sectional analysis of magnetic resonance images (MRI) in neurodegenerative diseases, namely Alzheimer's dementia (AD). In this paper, the accuracy of our ACC methodology is assessed when presented with real life, imperfect data, i.e., cohorts of MRI with varying acquisition parameters and imaging quality. The comparative methodology uses the Jacobian determinants derived from dense deformation fields and scaled grey-level intensity from a selected volume of interest centered on the medial temporal lobe. The ACC performance is assessed in a series of leave-one-out experiments aimed at separating 75 probable AD and 75 age-matched normal controls. The resulting accuracy is 92% using a support vector machine classifier based on least squares optimization. Finally, it is shown in the Appendix that determinants and scaled grey-level intensity are appreciably more robust to varying parameters in validation studies using simulated data, when compared to raw intensities or grey/white matter volumes. The ability of cross-sectional MRI at detecting probable AD with high accuracy could have profound implications in the management of suspected AD candidates. PMID- 18390348 TI - Comparison of residence time estimation methods for radioimmunotherapy dosimetry and treatment planning--Monte Carlo simulation studies. AB - Estimating the residence times in tumor and normal organs is an essential part of treatment planning for radioimmunotherapy (RIT). This estimation is usually done using a conjugate view whole body scan time series and planar processing. This method has logistical and cost advantages compared to 3-D imaging methods such as Single photon emission computed tomography (SPECT), but, because it does not provide information about the 3-D distribution of activity, it is difficult to fully compensate for effects such as attenuation and background and overlapping activity. Incomplete compensation for these effects reduces the accuracy of the residence time estimates. In this work we compare residence times estimates obtained using planar methods to those from methods based on quantitative SPECT (QSPECT) reconstructions. We have previously developed QSPECT methods that provide compensation for attenuation, scatter, collimator-detector response, and partial volume effects. In this study we compared the use of residence time estimation methods using QSPECT to planar methods. The evaluation was done using the realistic NCAT phantom with organ time activities that model (111)In ibritumomab tiuxetan. Projection data were obtained using Monte Carlo simulations (MCS) that realistically model the image formation process including penetration and scatter in the collimator-detector system. These projection data were used to evaluate the accuracy of residence time estimation using a time series of QSPECT studies, a single QSPECT study combined with planar scans and the planar scans alone. The errors in the residence time estimates were 3.8%, 15%, and 2%-107% for the QSPECT, hybrid planar/QSPECT, and planar methods, respectively. The quantitative accuracy was worst for pure planar processing and best for pure QSPECT processing. Hybrid planar/QSPECT methods, where a single QSPECT study was combined with a series of planar scans, provided a large and statistically significant improvement in quantitative accuracy for most organs compared to the planar scans alone, even without sophisticated attention to background subtraction or thickness corrections in planar processing. These results indicate that hybrid planar/QSPECT methods are generally superior to pure planar methods and may be an acceptable alternative to performing a time series of QSPECT studies. PMID- 18390349 TI - Accurate localization of brain activity in presurgical FMRI by structure adaptive smoothing. AB - An important problem of the analysis of functional magnetic resonance imaging (fMRI) experiments is to achieve some noise reduction of the data without blurring the shape of the activation areas. As a novel solution to this problem, recently the propagation-separation (PS) approach has been proposed. PS is a structure adaptive smoothing method that adapts to different shapes of activation areas. In this paper, we demonstrate how this method results in a more accurate localization of brain activity. First, it is shown in numerical simulations that PS is superior over Gaussian smoothing with respect to the accurate description of the shape of activation clusters and results in less false detections. Second, in a study of 37 presurgical planning cases we found that PS and Gaussian smoothing often yield different results, and we present examples showing aspects of the superiority of PS as applied to presurgical planning. PMID- 18390350 TI - Accelerating the nonequispaced fast Fourier transform on commodity graphics hardware. AB - We present a fast parallel algorithm to compute the nonequispaced fast Fourier transform on commodity graphics hardware (the GPU). We focus particularly on a novel implementation of the convolution step in the transform as it was previously its most time consuming part. We describe the performance for two common sample distributions in medical imaging (radial and spiral trajectories), and for different convolution kernels as these parameters all influence the speed of the algorithm. The GPU-accelerated convolution is up to 85 times faster as our reference, the open source NFFT library on a state-of-the-art 64 bit CPU. The accuracy of the proposed GPU implementation was quantitatively evaluated at the various settings. To illustrate the applicability of the transform in medical imaging, in which it is also known as gridding, we look specifically at non Cartesian magnetic resonance imaging and reconstruct both a numerical phantom and an in vivo cardiac image. PMID- 18390351 TI - Prostate cancer spectral multifeature analysis using TRUS images. AB - This paper focuses on extracting and analyzing different spectral features from transrectal ultrasound (TRUS) images for prostate cancer recognition. First, the information about the images' frequency domain features and spatial domain features are combined using a Gabor filter and then integrated with the expert radiologist's information to identify the highly suspicious regions of interest (ROIs). The next stage of the proposed algorithm is to scan each identified region in order to generate the corresponding 1-D signal that represents each region. For each ROI, possible spectral feature sets are constructed using different new geometrical features extracted from the power spectrum density (PSD) of each region's signal. Next, a classifier-based algorithm for feature selection using particle swarm optimization (PSO) is adopted and used to select the optimal feature subset from the constructed feature sets. A new spectral feature set for the TRUS images using estimation of signal parameters via rotational invariance technique (ESPRIT) is also constructed, and its ability to represent tissue texture is compared to the PSD-based spectral feature sets using the support vector machines (SVMs) classifier. The accuracy obtained ranges from 72.2% to 94.4%, with the best accuracy achieved by the ESPRIT feature set. PMID- 18390352 TI - Geometric variability of the scoliotic spine using statistics on articulated shape models. AB - This paper introduces a method to analyze the variability of the spine shape and of the spine shape deformations using articulated shape models. The spine shape was expressed as a vector of relative poses between local coordinate systems of neighboring vertebrae. Spine shape deformations were then modeled by a vector of rigid transformations that transforms one spine shape into another. Because rigid transformations do not naturally belong to a vector space, conventional mean and covariance could not be applied. The Frechet mean and a generalized covariance were used instead. The spine shapes of a group of 295 scoliotic patients were quantitatively analyzed as well as the spine shape deformations associated with the Cotrel-Dubousset corrective surgery (33 patients), the Boston brace (39 patients), and the scoliosis progression without treatment (26 patients). The variability of intervertebral poses was found to be inhomogeneous (lumbar vertebrae were more variable than the thoracic ones) and anisotropic (with maximal rotational variability around the coronal axis and maximal translational variability along the axial direction). Finally, brace and surgery were found to have a significant effect on the Frechet mean and on the generalized covariance in specific spine regions where treatments modified the spine shape. PMID- 18390353 TI - Reproducibility of global and local reconstruction of three-dimensional micro computed tomography of iliac crest biopsies. AB - Variation in computed tomography (CT) image gray-scale and spatial geometry due to specimen orientation, magnification, voxel size, differences in X-ray photon energy and limited field-of-view during the scan, were evaluated in repeated micro-CT scans of iliac crest biopsies and test phantoms. Using the micro-CT scanner on beamline X2B at the Brookhaven National Laboratory's National Synchrotron Light Source, 3-D micro-CT images were generated. They consisted of up to 1024 x 2400(2), 4-microm cubic voxels, each with 16-bit gray-scale. We also reconstructed the images at 16-, 32-, and 48-microm voxel resolution. Scan data were reconstructed from the complete profiles using filtered back-projection and from truncated profiles using profile-extension and with a Local reconstruction algorithm. Three biopsies and one bone-like test phantom were each rescanned at three different times at annual intervals. For the full-data-set reconstructions, the reproducibility of the estimates of mineral content of bone at mean bone opacity value, was +/-28.8, i.e., 2.56%, in a 4-microm cubic voxel at the 95% confidence level. The reproducibility decreased with increased voxel size. The interscan difference in imaged bone volume ranged from 0.86 4-microm 0.64% at 4 microm voxel resolution, and 2.64 4-microm 2.48% at 48 microm. PMID- 18390354 TI - A discriminative approach for wavelet denoising. AB - This paper suggests a discriminative approach for wavelet denoising where a set of mapping functions (MFs) are applied to the transform coefficients in an attempt to produce a noise free image. As opposed to the descriptive approaches, modeling image or noise priors is not required here and the MFs are learned directly from an ensemble of example images using least-squares fitting. The suggested scheme generates a novel set of MFs that are essentially different from the traditional soft/hard thresholding in the over-complete case. These MFs are demonstrated to obtain comparable performance to the state-of-the-art denoising approaches. Additionally, this framework enables a seamless customization of the shrinkage operation to a new set of restoration problems that were not addressed previously with shrinkage techniques, such as deblurring, JPEG artifact removal, and various types of additive noise that are not necessarily Gaussian white noise. PMID- 18390355 TI - Maximum likelihood wavelet density estimation with applications to image and shape matching. AB - Density estimation for observational data plays an integral role in a broad spectrum of applications, e.g., statistical data analysis and information theoretic image registration. Of late, wavelet-based density estimators have gained in popularity due to their ability to approximate a large class of functions, adapting well to difficult situations such as when densities exhibit abrupt changes. The decision to work with wavelet density estimators brings along with it theoretical considerations (e.g., non-negativity, integrability) and empirical issues (e.g., computation of basis coefficients) that must be addressed in order to obtain a bona fide density. In this paper, we present a new method to accurately estimate a non-negative density which directly addresses many of the problems in practical wavelet density estimation. We cast the estimation procedure in a maximum likelihood framework which estimates the square root of the density radicalp, allowing us to obtain the natural non-negative density representation ( radicalp)(2). Analysis of this method will bring to light a remarkable theoretical connection with the Fisher information of the density and, consequently, lead to an efficient constrained optimization procedure to estimate the wavelet coefficients. We illustrate the effectiveness of the algorithm by evaluating its performance on mutual information-based image registration, shape point set alignment, and empirical comparisons to known densities. The present method is also compared to fixed and variable bandwidth kernel density estimators. PMID- 18390356 TI - Wavelet-based Bayesian image estimation: from marginal and bivariate prior models to multivariate prior models. AB - Prior models play an important role in the wavelet-based Bayesian image estimation problem. Although it is well known that a residual dependency structure always remains among natural image wavelet coefficients, only few multivariate prior models with a closed parametric form are available in the literature. In this paper, we develop new multivariate prior models that not only match well with the observed statistics of the wavelet coefficients of natural images, but also have a simple parametric form. These prior models are very effective for Bayesian image estimation and lead to an improved estimation performance over related earlier techniques. PMID- 18390357 TI - SURE-LET multichannel image denoising: interscale orthonormal wavelet thresholding. AB - We propose a vector/matrix extension of our denoising algorithm initially developed for grayscale images, in order to efficiently process multichannel (e.g., color) images. This work follows our recently published SURE-LET approach where the denoising algorithm is parameterized as a linear expansion of thresholds (LET) and optimized using Stein's unbiased risk estimate (SURE). The proposed wavelet thresholding function is pointwise and depends on the coefficients of same location in the other channels, as well as on their parents in the coarser wavelet subband. A nonredundant, orthonormal, wavelet transform is first applied to the noisy data, followed by the (subband-dependent) vector valued thresholding of individual multichannel wavelet coefficients which are finally brought back to the image domain by inverse wavelet transform. Extensive comparisons with the state-of-the-art multiresolution image denoising algorithms indicate that despite being nonredundant, our algorithm matches the quality of the best redundant approaches, while maintaining a high computational efficiency and a low CPU/memory consumption. An online Java demo illustrates these assertions. PMID- 18390358 TI - Nonrigid registration of 3-d multichannel microscopy images of cell nuclei. AB - We present an intensity-based nonrigid registration approach for the normalization of 3-D multichannel microscopy images of cell nuclei. A main problem with cell nuclei images is that the intensity structure of different nuclei differs very much; thus, an intensity-based registration scheme cannot be used directly. Instead, we first perform a segmentation of the images from the cell nucleus channel, smooth the resulting images by a Gaussian filter, and then apply an intensity-based registration algorithm. The obtained transformation is applied to the images from the nucleus channel as well as to the images from the other channels. To improve the convergence rate of the algorithm, we propose an adaptive step length optimization scheme and also employ a multiresolution scheme. Our approach has been successfully applied using 2-D cell-like synthetic images, 3-D phantom images as well as 3-D multichannel microscopy images representing different chromosome territories and gene regions. We also describe an extension of our approach, which is applied for the registration of 3D + t (4 D) image series of moving cell nuclei. PMID- 18390359 TI - Weighted adaptive lifting-based wavelet transform for image coding. AB - In this paper, a new weighted adaptive lifting (WAL)-based wavelet transform is presented. The proposed WAL approach is designed to solve the problems existing in the previous adaptive directional lifting (ADL) approach, such as mismatch between the predict and update steps, interpolation favoring only horizontal or vertical direction, and invariant interpolation filter coefficients for all images. The main contribution of the proposed approach consists of two parts: one is the improved weighted lifting, which maintains the consistency between the predict and update steps as far as possible and preserves the perfect reconstruction at the same time; another is the directional adaptive interpolation, which improves the orientation property of the interpolated image and adapts to statistical property of each image. Experimental results show that the proposed WAL-based wavelet transform for image coding outperforms the conventional lifting-based wavelet transform up to 3.06 dB in PSNR and significant improvement in subjective quality is also observed. Compared with the ADL-based wavelet transform, up to 1.22-dB improvement in PSNR is reported. PMID- 18390360 TI - Universal image compression using multiscale recurrent patterns with adaptive probability model. AB - In this work, we further develop the multidimensional multiscale parser (MMP) algorithm, a recently proposed universal lossy compression method which has been successfully applied to images as well as other types of data, as video and ECG signals. The MMP is based on approximate multiscale pattern matching, encoding blocks of an input signal using expanded and contracted versions of patterns stored in a dictionary. The dictionary is updated using expanded and contracted versions of concatenations of previously encoded blocks. This implies that MMP builds its own dictionary while the input data is being encoded, using segments of the input itself, which lends it a universal flavor. It presents a flexible structure, which allows for easily adding data-specific extensions to the base algorithm. Often, the signals to be encoded belong to a narrow class, as the one of smooth images. In these cases, one expects that some improvement can be achieved by introducing some knowledge about the source to be encoded. In this paper, we use the assumption about the smoothness of the source in order to create good context models for the probability of blocks in the dictionary. Such probability models are estimated by considering smoothness constraints around causal block boundaries. In addition, we refine the obtained probability models by also exploiting the existing knowledge about the original scale of the included blocks during the dictionary updating process. Simulation results have shown that these developments allow significant improvements over the original MMP for smooth images, while keeping its state-of-the-art performance for more complex, less smooth ones, thus improving MMP's universal character. PMID- 18390361 TI - Fast full-search equivalent template matching by enhanced bounded correlation. AB - We propose a novel algorithm, referred to as enhanced bounded correlation (EBC), that significantly reduces the number of computations required to carry out template matching based on normalized cross correlation (NCC) and yields exactly the same result as the full search algorithm. The algorithm relies on the concept of bounding the matching function: finding an efficiently computable upper bound of the NCC rapidly prunes those candidates that cannot provide a better NCC score with respect to the current best match. In this framework, we apply a succession of increasingly tighter upper bounding functions based on Cauchy-Schwarz inequality. Moreover, by including an online parameter prediction step into EBC, we obtain a parameter free algorithm that, in most cases, affords computational advantages very similar to those attainable by optimal offline parameter tuning. Experimental results show that the proposed algorithm can significantly accelerate a full-search equivalent template matching process and outperforms state-of-the-art methods. PMID- 18390362 TI - A fast thresholded landweber algorithm for wavelet-regularized multidimensional deconvolution. AB - We present a fast variational deconvolution algorithm that minimizes a quadratic data term subject to a regularization on the l(1)-norm of the wavelet coefficients of the solution. Previously available methods have essentially consisted in alternating between a Landweber iteration and a wavelet-domain soft thresholding operation. While having the advantage of simplicity, they are known to converge slowly. By expressing the cost functional in a Shannon wavelet basis, we are able to decompose the problem into a series of subband-dependent minimizations. In particular, this allows for larger (subband-dependent) step sizes and threshold levels than the previous method. This improves the convergence properties of the algorithm significantly. We demonstrate a speed-up of one order of magnitude in practical situations. This makes wavelet-regularized deconvolution more widely accessible, even for applications with a strong limitation on computational complexity. We present promising results in 3-D deconvolution microscopy, where the size of typical data sets does not permit more than a few tens of iterations. PMID- 18390363 TI - Deblurring using regularized locally adaptive kernel regression. AB - Kernel regression is an effective tool for a variety of image processing tasks such as denoising and interpolation [1]. In this paper, we extend the use of kernel regression for deblurring applications. In some earlier examples in the literature, such nonparametric deblurring was suboptimally performed in two sequential steps, namely denoising followed by deblurring. In contrast, our optimal solution jointly denoises and deblurs images. The proposed algorithm takes advantage of an effective and novel image prior that generalizes some of the most popular regularization techniques in the literature. Experimental results demonstrate the effectiveness of our method. PMID- 18390364 TI - GAFFE: a gaze-attentive fixation finding engine. AB - The ability to automatically detect visually interesting regions in images has many practical applications, especially in the design of active machine vision and automatic visual surveillance systems. Analysis of the statistics of image features at observers' gaze can provide insights into the mechanisms of fixation selection in humans. Using a foveated analysis framework, we studied the statistics of four low-level local image features: luminance, contrast, and bandpass outputs of both luminance and contrast, and discovered that image patches around human fixations had, on average, higher values of each of these features than image patches selected at random. Contrast-bandpass showed the greatest difference between human and random fixations, followed by luminance bandpass, RMS contrast, and luminance. Using these measurements, we present a new algorithm that selects image regions as likely candidates for fixation. These regions are shown to correlate well with fixations recorded from human observers. PMID- 18390365 TI - Topology preserving non-negative matrix factorization for face recognition. AB - In this paper, a novel topology preserving non-negative matrix factorization (TPNMF) method is proposed for face recognition. We derive the TPNMF model from original NMF algorithm by preserving local topology structure. The TPNMF is based on minimizing the constraint gradient distance in the high-dimensional space. Compared with L(2) distance, the gradient distance is able to reveal latent manifold structure of face patterns. By using TPNMF decomposition, the high dimensional face space is transformed into a local topology preserving subspace for face recognition. In comparison with PCA, LDA, and original NMF, which search only the Euclidean structure of face space, the proposed TPNMF finds an embedding that preserves local topology information, such as edges and texture. Theoretical analysis and derivation given also validate the property of TPNMF. Experimental results on three different databases, containing more than 12,000 face images under varying in lighting, facial expression, and pose, show that the proposed TPNMF approach provides a better representation of face patterns and achieves higher recognition rates than NMF. PMID- 18390366 TI - Snakuscules. AB - A snakuscule (a minuscule snake) is the simplest active contour that we were able to design while keeping the quintessence of traditional snakes: an energy term governed by the data, and a regularization term. Our construction is an area based snake, as opposed to curve-based snakes. It is parameterized by just two points, thus further easing requirements on the optimizer. Despite their ultimate simplicity, snakuscules retain enough versatility to be employed for solving various problems such as cell counting and segmentation of approximately circular features. In this paper, we detail the design process of a snakuscule and illustrate its usefulness through practical examples. We claim that our didactic intentions are well served by the simplicity of snakuscules. PMID- 18390367 TI - Activity modeling using event probability sequences. AB - Changes in motion properties of trajectories provide useful cues for modeling and recognizing human activities. We associate an event with significant changes that are localized in time and space, and represent activities as a sequence of such events. The localized nature of events allows for detection of subtle changes or anomalies in activities. In this paper, we present a probabilistic approach for representing events using the hidden Markov model (HMM) framework. Using trained HMMs for activities, an event probability sequence is computed for every motion trajectory in the training set. It reflects the probability of an event occurring at every time instant. Though the parameters of the trained HMMs depend on viewing direction, the event probability sequences are robust to changes in viewing direction. We describe sufficient conditions for the existence of view invariance. The usefulness of the proposed event representation is illustrated using activity recognition and anomaly detection. Experiments using the indoor University of Central Florida human action dataset, the Carnegie Mellon University Credo Intelligence, Inc., Motion Capture dataset, and the outdoor Transportation Security Administration airport tarmac surveillance dataset show encouraging results. PMID- 18390368 TI - Bayesian foreground and shadow detection in uncertain frame rate surveillance videos. AB - In in this paper, we propose a new model regarding foreground and shadow detection in video sequences. The model works without detailed a priori object shape information, and it is also appropriate for low and unstable frame rate video sources. Contribution is presented in three key issues: 1) we propose a novel adaptive shadow model, and show the improvements versus previous approaches in scenes with difficult lighting and coloring effects; 2) we give a novel description for the foreground based on spatial statistics of the neighboring pixel values, which enhances the detection of background or shadow-colored object parts; 3) we show how microstructure analysis can be used in the proposed framework as additional feature components improving the results. Finally, a Markov random field model is used to enhance the accuracy of the separation. We validate our method on outdoor and indoor sequences including real surveillance videos and well-known benchmark test sets. PMID- 18390369 TI - Customizing kernel functions for SVM-based hyperspectral image classification. AB - Previous research applying kernel methods such as support vector machines (SVMs) to hyperspectral image classification has achieved performance competitive with the best available algorithms. However, few efforts have been made to extend SVMs to cover the specific requirements of hyperspectral image classification, for example, by building tailor-made kernels. Observation of real-life spectral imagery from the AVIRIS hyperspectral sensor shows that the useful information for classification is not equally distributed across bands, which provides potential to enhance the SVM's performance through exploring different kernel functions. Spectrally weighted kernels are, therefore, proposed, and a set of particular weights is chosen by either optimizing an estimate of generalization error or evaluating each band's utility level. To assess the effectiveness of the proposed method, experiments are carried out on the publicly available 92AV3C dataset collected from the 220-dimensional AVIRIS hyperspectral sensor. Results indicate that the method is generally effective in improving performance: spectral weighting based on learning weights by gradient descent is found to be slightly better than an alternative method based on estimating "relevance" between band information and ground truth. PMID- 18390370 TI - Fast and robust generation of feature maps for region-based visual attention. AB - Visual attention is one of the important phenomena in biological vision which can be followed to achieve more efficiency, intelligence, and robustness in artificial vision systems. This paper investigates a region-based approach that performs pixel clustering prior to the processes of attention in contrast to late clustering as done by contemporary methods. The foundation steps of feature map construction for the region-based attention model are proposed here. The color contrast map is generated based upon the extended findings from the color theory, the symmetry map is constructed using a novel scanning-based method, and a new algorithm is proposed to compute a size contrast map as a formal feature channel. Eccentricity and orientation are computed using the moments of obtained regions and then saliency is evaluated using the rarity criteria. The efficient design of the proposed algorithms allows incorporating five feature channels while maintaining a processing rate of multiple frames per second. Another salient advantage over the existing techniques is the reusability of the salient regions in the high-level machine vision procedures due to preservation of their shapes and precise locations. The results indicate that the proposed model has the potential to efficiently integrate the phenomenon of attention into the main stream of machine vision and systems with restricted computing resources such as mobile robots can benefit from its advantages. PMID- 18390371 TI - A real-time algorithm for the approximation of level-set-based curve evolution. AB - In this paper, we present a complete and practical algorithm for the approximation of level-set-based curve evolution suitable for real-time implementation. In particular, we propose a two-cycle algorithm to approximate level-set-based curve evolution without the need of solving partial differential equations (PDEs). Our algorithm is applicable to a broad class of evolution speeds that can be viewed as composed of a data-dependent term and a curve smoothness regularization term. We achieve curve evolution corresponding to such evolution speeds by separating the evolution process into two different cycles: one cycle for the data-dependent term and a second cycle for the smoothness regularization. The smoothing term is derived from a Gaussian filtering process. In both cycles, the evolution is realized through a simple element switching mechanism between two linked lists, that implicitly represents the curve using an integer valued level-set function. By careful construction, all the key evolution steps require only integer operations. A consequence is that we obtain significant computation speedups compared to exact PDE-based approaches while obtaining excellent agreement with these methods for problems of practical engineering interest. In particular, the resulting algorithm is fast enough for use in real-time video processing applications, which we demonstrate through several image segmentation and video tracking experiments. PMID- 18390372 TI - A wavelet-laplace variational technique for image deconvolution and inpainting. AB - We construct a new variational method for blind deconvolution of images and inpainting, motivated by recent PDE-based techniques involving the Ginzburg Landau functional, but using more localized wavelet-based methods. We present results for both binary and grayscale images. Comparable speeds are achieved with better sharpness of edges in the reconstruction. PMID- 18390373 TI - Adaptive bilateral filter for sharpness enhancement and noise removal. AB - In this paper, we present the adaptive bilateral filter (ABF) for sharpness enhancement and noise removal. The ABF sharpens an image by increasing the slope of the edges without producing overshoot or undershoot. It is an approach to sharpness enhancement that is fundamentally different from the unsharp mask (USM). This new approach to slope restoration also differs significantly from previous slope restoration algorithms in that the ABF does not involve detection of edges or their orientation, or extraction of edge profiles. In the ABF, the edge slope is enhanced by transforming the histogram via a range filter with adaptive offset and width. The ABF is able to smooth the noise, while enhancing edges and textures in the image. The parameters of the ABF are optimized with a training procedure. ABF restored images are significantly sharper than those restored by the bilateral filter. Compared with an USM based sharpening method the optimal unsharp mask (OUM), ABF restored edges are as sharp as those rendered by the OUM, but without the halo artifacts that appear in the OUM restored image. In terms of noise removal, ABF also outperforms the bilateral filter and the OUM. We demonstrate that ABF works well for both natural images and text images. PMID- 18390374 TI - Reversible, fast, and high-quality grid conversions. AB - A new grid conversion method is proposed to resample between two 2-D periodic lattices with the same sampling density. The main feature of our approach is the symmetric reversibility, which means that when using the same algorithm for the converse operation, then the initial data is recovered exactly. To that purpose, we decompose the lattice conversion process into (at most) three successive shear operations. The translations along the shear directions are implemented by 1-D fractional delay operators, which revert to simple 1-D convolutions, with appropriate filters that yield the property of symmetric reversibility. We show that the method is fast and provides high-quality resampled images. Applications of our approach can be found in various settings, such as grid conversion between the hexagonal and the Cartesian lattice, or fast implementation of affine transformations such as rotations. PMID- 18390375 TI - A multistage motion vector processing method for motion-compensated frame interpolation. AB - In this paper, a novel, low-complexity motion vector processing algorithm at the decoder is proposed for motion-compensated frame interpolation or frame rate up conversion. We address the problems of having broken edges and deformed structures in an interpolated frame by hierarchically refining motion vectors on different block sizes. Our method explicitly considers the reliability of each received motion vector and has the capability of preserving the structure information. This is achieved by analyzing the distribution of residual energies and effectively merging blocks that have unreliable motion vectors. The motion vector reliability information is also used as a prior knowledge in motion vector refinement using a constrained vector median filter to avoid choosing identical unreliable one. We also propose using chrominance information in our method. Experimental results show that the proposed scheme has better visual quality and is also robust, even in video sequences with complex scenes and fast motion. PMID- 18390376 TI - RDTC optimized compression of image-based scene representations (Part I): modeling and theoretical analysis. AB - Rendering of virtual views in interactive streaming of compressed image-based scene representations requires random access to arbitrary parts of the reference image data. The degree of interframe dependencies exploited during encoding has an impact on the transmission and decoding time and, at the same time, delimits the (storage) rate-distortion (RD) tradeoff that can be achieved. In this work, we extend the classical RD optimization approach using hybrid video coding concepts to a tradeoff between the storage rate (R), distortion (D), transmission data rate (T), and decoding complexity (C). We present a theoretical model for this RDTC space with a focus on the decoding complexity and, in addition, the impact of client side caching on the RDTC measures is considered and evaluated. Experimental results qualitatively match those predicted by our theoretical models and show that an adaptation of the encoding process to scenario specific parameters like computational power of the receiver and channel throughput can significantly reduce the user-perceived delay or required storage for RDTC optimized streams compared to RD optimized or independently encoded scene representations. PMID- 18390377 TI - RDTC optimized compression of image-based scene representations (Part II): practical coding. AB - Interactive streaming of compressed image-based scene representations requires random access to the reference image data. The degree of interframe dependencies exploited during encoding has an impact on the transmission and decoding time and, at the same time, delimits the (storage) rate-distortion (RD) tradeoff that can be achieved. The transmission data rate and the decoding complexity at the client have received attention in the literature, but their incorporation into the optimization procedure for compression and streaming is missing. If scenario specific measures are considered, the traditional RD optimization can be extended to a tradeoff between the (storage) rate (R), distortion (D), transmission data rate (T), and decoding complexity (C). In the first part of this sequel of papers, we have theoretically analyzed the RDTC space for the compression of densely sampled image-based scene representations. In this second part, we consider practical RDTC optimization. We propose a modeling and encoding parameter selection procedure that allows us to adapt the compression to scenario specific properties. The impact of client side caching is considered and evaluated using an experimental testbed. Our results show a significant reduction of the user perceived delay, memory consumption or required minimum channel and storage bitrate for RDTC optimized streams compared to classical RD optimized or independently encoded scene representations. PMID- 18390378 TI - Algorithmic and architectural optimizations for computationally efficient particle filtering. AB - In this paper, we analyze the computational challenges in implementing particle filtering, especially to video sequences. Particle filtering is a technique used for filtering nonlinear dynamical systems driven by non-Gaussian noise processes. It has found widespread applications in detection, navigation, and tracking problems. Although, in general, particle filtering methods yield improved results, it is difficult to achieve real time performance. In this paper, we analyze the computational drawbacks of traditional particle filtering algorithms, and present a method for implementing the particle filter using the Independent Metropolis Hastings sampler, that is highly amenable to pipelined implementations and parallelization. We analyze the implementations of the proposed algorithm, and, in particular, concentrate on implementations that have minimum processing times. It is shown that the design parameters for the fastest implementation can be chosen by solving a set of convex programs. The proposed computational methodology was verified using a cluster of PCs for the application of visual tracking. We demonstrate a linear speed-up of the algorithm using the methodology proposed in the paper. PMID- 18390379 TI - A run-based two-scan labeling algorithm. AB - We present an efficient run-based two-scan algorithm for labeling connected components in a binary image. Unlike conventional label-equivalence-based algorithms, which resolve label equivalences between provisional labels, our algorithm resolves label equivalences between provisional label sets. At any time, all provisional labels that are assigned to a connected component are combined in a set, and the smallest label is used as the representative label. The corresponding relation of a provisional label and its representative label is recorded in a table. Whenever different connected components are found to be connected, all provisional label sets concerned with these connected components are merged together, and the smallest provisional label is taken as the representative label. When the first scan is finished, all provisional labels that were assigned to each connected component in the given image will have a unique representative label. During the second scan, we need only to replace each provisional label by its representative label. Experimental results on various types of images demonstrate that our algorithm outperforms all conventional labeling algorithms. PMID- 18390380 TI - Unified anomaly suppression and boundary extraction in laser radar range imagery based on a joint curve-evolution and expectation-maximization algorithm. AB - In this paper, we develop a new unified approach for laser radar range anomaly suppression, range profiling, and segmentation. This approach combines an object based hybrid scene model for representing the range distribution of the field and a statistical mixture model for the range data measurement noise. The image segmentation problem is formulated as a minimization problem which jointly estimates the target boundary together with the target region range variation and background range variation directly from the noisy and anomaly-filled range data. This formulation allows direct incorporation of prior information concerning the target boundary, target ranges, and background ranges into an optimal reconstruction process. Curve evolution techniques and a generalized expectation maximization algorithm are jointly employed as an efficient solver for minimizing the objective energy, resulting in a coupled pair of object and intensity optimization tasks. The method directly and optimally extracts the target boundary, avoiding a suboptimal two-step process involving image smoothing followed by boundary extraction. Experiments are presented demonstrating that the proposed approach is robust to anomalous pixels (missing data) and capable of producing accurate estimation of the target boundary and range values from noisy data. PMID- 18390381 TI - Self-repelling snakes for topology-preserving segmentation models. AB - The implicit framework of the level-set method has several advantages when tracking propagating fronts. Indeed, the evolving contour is embedded in a higher dimensional level-set function and its evolution can be phrased in terms of a Eulerian formulation. The ability of this intrinsic method to handle topological changes (merging and breaking) makes it useful in a wide range of applications (fluid mechanics, computer vision) and particularly in image segmentation, the main subject of this paper. Nevertheless, in some applications, this topological flexibility turns out to be undesirable: for instance, when the shape to be detected has a known topology, or when the resulting shape must be homeomorphic to the initial one. The necessity of designing topology-preserving processes arises in medical imaging, for example, in the human cortex reconstruction. It is known that the human cortex has a spherical topology so throughout the reconstruction process this topological feature must be preserved. Therefore, we propose in this paper a segmentation model based on an implicit level-set formulation and on the geodesic active contours, in which a topological constraint is enforced. PMID- 18390382 TI - Segmentation by fusion of histogram-based k-means clusters in different color spaces. AB - This paper presents a new, simple, and efficient segmentation approach, based on a fusion procedure which aims at combining several segmentation maps associated to simpler partition models in order to finally get a more reliable and accurate segmentation result. The different label fields to be fused in our application are given by the same and simple (K-means based) clustering technique on an input image expressed in different color spaces. Our fusion strategy aims at combining these segmentation maps with a final clustering procedure using as input features, the local histogram of the class labels, previously estimated and associated to each site and for all these initial partitions. This fusion framework remains simple to implement, fast, general enough to be applied to various computer vision applications (e.g., motion detection and segmentation), and has been successfully applied on the Berkeley image database. The experiments herein reported in this paper illustrate the potential of this approach compared to the state-of-the-art segmentation methods recently proposed in the literature. PMID- 18390383 TI - Using spanning graphs for efficient image registration. AB - We provide a detailed analysis of the use of minimal spanning graphs as an alignment method for registering multimodal images. This yields an efficient graph theoretic algorithm that, for the first time, jointly estimates both an alignment measure and a viable descent direction with respect to a parameterized class of spatial transformations. We also show how prior information about the interimage modality relationship from prealigned image pairs can be incorporated into the graph-based algorithm. A comparison of the graph theoretic alignment measure is provided with more traditional measures based on plug-in entropy estimators. This highlights previously unrecognized similarities between these two registration methods. Our analysis gives additional insight into the tradeoffs the graph-based algorithm is making and how these will manifest themselves in the registration algorithm's performance. PMID- 18390384 TI - Block-coordinate Gauss-Newton optimization and constrained monotone regression for image registration in the presence of outlier objects. AB - In this paper, we propose the block-coordinate Gauss- Newton/regression method in order to conduct a correlation-based registration considering the intensity difference between images in the presence of outlier objects. In the proposed method, the parameters are decomposed into two blocks, one of which is for the spatial registration and the other for the intensity compensation. The two blocks are sequentially updated by the Gauss-Newton update and the polynomial regression, respectively. Because of the separated blocks, we can perform a joint optimization with low computational complexity and high implementation flexibility. For example, we apply separately appropriate scaling techniques to the parameter blocks for a stable and fast convergence of the algorithm. Furthermore, we apply the constrained monotone regression with a robust outlier detection scheme for the intensity compensation block. From numerical results, it is shown that the proposed algorithm more effectively performs a correlation based registration considering the intensity difference alleviating the influence of the outlier objects compared to the traditional registration algorithms that perform the joint optimization. PMID- 18390385 TI - Infinite hidden Markov models for unusual-event detection in video. AB - We address the problem of unusual-event detection in a video sequence. Invariant subspace analysis (ISA) is used to extract features from the video, and the time evolving properties of these features are modeled via an infinite hidden Markov model (iHMM), which is trained using "normal"/"typical" video. The iHMM retains a full posterior density function on all model parameters, including the number of underlying HMM states. Anomalies (unusual events) are detected subsequently if a low likelihood is observed when associated sequential features are submitted to the trained iHMM. A hierarchical Dirichlet process framework is employed in the formulation of the iHMM. The evaluation of posterior distributions for the iHMM is achieved in two ways: via Markov chain Monte Carlo and using a variational Bayes formulation. Comparisons are made to modeling based on conventional maximum likelihood-based HMMs, as well as to Dirichlet-process-based Gaussian-mixture models. PMID- 18390386 TI - Continuous sampling in mutual-information registration. AB - Mutual information is a popular and widely used metric in retrospective image registration. This metric excels especially with multimodal data due to the minimal assumptions about the correspondence between the image intensities. In certain situations, the mutual-information metric is known to produce artifacts that rule out subsample registration accuracy. Various methods have been developed to mitigate these artifacts, including higher order kernels for smoother sampling of the metric. This study introduces a novel concept of continuous sampling to provide new insight into the mutual-information methods currently in use. In particular, the connection between the partial volume interpolation and the recently introduced higher order partial-volume-type kernels is revealed. PMID- 18390387 TI - Magnetic resonance imaging features of renal synovial sarcoma: a case report. AB - Primary renal synovial sarcoma (SS) was first described in 2000 by Argani, with only a few subsequent cases being reported in the English literature. Herein, we describe a case of a 52-year-old woman who presented with right flank pain. Magnetic resonance imaging revealed a 6-cm mass in the lower pole of the right kidney. T1 and T2 weighted imaging revealed a heterogeneous mass with triple sign. There was post-contrast enhancement. Imaging, histology and immunostaining together made the diagnosis of SS of the kidney. PMID- 18390388 TI - Ultrasound of malignant cervical lymph nodes. AB - Malignant lymph nodes in the neck include metastases and lymphoma. Cervical nodal metastases are common in patients with head and neck cancers, and their assessment is important as it affects treatment planning and prognosis. Neck nodes are also a common site of lymphomatous involvement and an accurate diagnosis is essential as its treatment differs from other causes of neck lymphadenopathy. On ultrasound, grey scale sonography helps to evaluate nodal morphology, whilst power Doppler sonography is used to assess the vascular pattern. Grey scale sonographic features that help to identify metastatic and lymphomatous lymph nodes include size, shape and internal architecture (loss of hilar architecture, presence of intranodal necrosis and calcification). Soft tissue oedema and nodal matting are additional grey scale features seen in tuberculous nodes or in nodes that have been previously irradiated. Power Doppler sonography evaluates the vascular pattern of nodes and helps to identify the malignant nodes. In addition, serial monitoring of nodal size and vascularity are useful features in the assessment of treatment response. PMID- 18390389 TI - Imaging for staging and management of thyroid cancer. AB - The management of thyroid cancer has been controversial and, as a result, the routine use of imaging in this disease, especially for pre-operative staging, has lagged behind other head and neck cancers. However, as more is known about the natural history of thyroid cancer, the role of imaging is becoming more established. This review focuses on how imaging now influences the staging and management of the primary cancer, nodal metastases and distant metastases. This is followed by a brief review of the role of imaging in planning post-operative radiotherapy and post-treatment surveillance. PMID- 18390391 TI - Warburg revisited: imaging tumour blood flow and metabolism. AB - In the 1930s, Otto Warburg reported that anaerobic metabolism of glucose is a fundamental property of all tumours, even in the presence of an adequate oxygen supply. He also demonstrated a relationship between the degree of anaerobic metabolism and tumour growth rate. Today, this phenomenon forms the basis of tumour imaging with fluorodeoxyglucose positron emission tomography (FDG-PET). More recently, Folkman has demonstrated that malignant growth and survival are also dependent on tumour vascularity which is increasingly evaluated in vivo using techniques such as contrast enhanced computed tomography or magnetic resonance imaging (MRI). Although it is reasonable to hypothesise that the metabolic requirements of tumours are mirrored by alterations in tumour haemodynamics, the relationship between tumour blood flow and metabolism is in fact complex. A well-developed tumour vascular supply is required to ensure a sufficient delivery of glucose and oxygen to support the metabolism essential for tumour growth. However, an inadequate vascularisation of tumour will result in hypoxia, a factor that is known to stimulate anaerobic metabolism of glucose. Thus, the balance between tumour blood flow and metabolism will be an important indicator of the biological status of a tumour and hence the tumour's likely progression and response to treatment. This article reviews the molecular biology of tumour vascularisation and metabolism, relating these processes to currently available imaging techniques while summarising the imaging studies that have compared tumour blood flow and metabolism. The potential for vascular metabolic imaging to assess tumour aggression and sub-classify treatment response is highlighted. PMID- 18390392 TI - New frontiers in pediatric oncologic imaging. AB - As imaging technologies advance, a paradigm shift is emerging in the assessment of tumor response to therapy. The traditional method of measuring tumor size may not reflect changes in tumor viability induced by chemotherapy and radiation therapy. Today's oncologists and radiologists seek objective methods for assessing tumor metabolism and blood flow, measures that provide earlier, more accurate information about treatment effects. Pediatric imaging presents unique challenges not encountered in adult imaging, including the need for sedation and consideration of the long-term effects of radiation exposure in a growing child. Therefore, the potential risks and benefits of new imaging approaches for monitoring anticancer treatment in children require careful consideration. Several new imaging techniques are currently under investigation for use in pediatric oncology. These include dynamic enhanced magnetic resonance imaging and quantitative contrast-enhanced ultrasonography for assessment of blood flow in solid tumors such as osteosarcoma and neuroblastoma, and nuclear imaging, including positron emission tomography-computed tomography, for assessment of pediatric musculoskeletal tumors and neuroblastoma. The potential value, relative advantages, and limitations of these new methods in monitoring anticancer therapy in children are discussed. PMID- 18390393 TI - A big step toward becoming an SCI journal. PMID- 18390390 TI - Predictive and prognostic value of FDG-PET. AB - The predictive and prognostic value of fluorodeoxyglucose (FDG)-positron emission tomography (PET) in non-small-cell lung carcinoma, colorectal carcinoma and lymphoma is discussed. The degree of FDG uptake is of prognostic value at initial presentation, after induction treatment prior to resection and in the case of relapse of non-small cell lung cancer (NSCLC). In locally advanced and advanced stages of NSCLC, FDG-PET has been shown to be predictive for clinical outcome at an early stage of treatment. In colorectal carcinoma, limited studies are available on the prognostic value of FDG-PET, however, the technique appears to have great potential in monitoring the success of local ablative therapies soon after intervention and in the prediction and evaluation of response to radiotherapy, systemic therapy, and combinations thereof. The prognostic value of end-of treatment FDG-PET for FDG-avid lymphomas has been established, and the next step is to define how to use this information to optimize patient outcome. In Hodgkin's lymphoma, FDG-PET has a high negative predictive value, however, histological confirmation of positive findings should be sought where possible. For non-Hodgkin's lymphoma, the opposite applies. The newly published standardized guidelines for interpretation formulates specific criteria for visual interpretation and for defining PET positivity in the liver, spleen, lung, bone marrow and small residual lesions. The introduction of these guidelines should reduce variability among studies. Interim PET offers a reliable method for early prediction of long-term remission, however it should only be performed in prospective randomized controlled trials. Many of the diagnostic and management questions considered in this review are relevant to other tumour types. Further research in this field is of great importance, since it may lead to a change in the therapeutic concept of cancer. The preliminary findings call for systematic inclusion of FDG-PET in therapeutic trials to adequately position FDG-PET in treatment time lines. PMID- 18390394 TI - Induction of hepatic glutathione S-transferase and UDP-glucuronosyltransferase activities by ketamine in rats. AB - BACKGROUND: Ketamine has been shown to induce rat cytochrome P-450 in a way similar to phenobarbital. However, whether ketamine is able to induce glutathione S-transferase (GST) and UDP-glucuronosyltransferase (UGT), two major phase II drug-metabolizing enzymes, remains unclear. The present study aimed to investigate the effect of ketamine on GST and UGT activities in rats. METHODS: In a dose-response study, male adult Wistar rats were treated with 10, 20, 40 or 80 mg/kg ketamine intraperitoneally twice daily for 4 days. Livers were removed 1 day after ketamine treatment and hepatic GST and UGT activities were determined. In a reversibility study, rats were treated with 80 mg/kg ketamine intraperitoneally twice daily for 4 days and killed 1, 2, 3 or 4 days after the last dose of ketamine. Livers were removed and hepatic GST and UGT activities were determined. RESULTS: The results of the dose-response study showed that treatment of rats with 10, 20, 40, or 80 mg/kg ketamine produced 19%, 20%, 18%, and 25% increases respectively in the catalytic activity of hepatic cytosolic GST, and 41%, 41%, 35%, and 38% increases respectively in the catalytic activity of microsomal UGT. The results of the reversibility study showed that the GST activities of the rats killed 1, 2, 3, or 4 days after ketamine treatment were 62%, 88%, 46% and 65% higher than the activity of the control group. The UGT activities of the rats killed 1, 2, 3, or 4 days after ketamine treatment were 56%, 53%, 54% and 72% higher than the activity of the control group. CONCLUSION: Ketamine is able to induce the activities of hepatic GST and UGT in rats. The induced GST and UGT activities persist for at least 4 days after cessation of ketamine. The results suggest the possibility of interactions of drugs related to phase II enzyme induction in chronic ketamine users. PMID- 18390395 TI - Hemodynamic responses to tracheal intubation with the Glidescope videolaryngoscope: a comparison of oral and nasal routes. AB - BACKGROUND: Although there have been numerous favorable reports on the uses of the GlideScope videolaryngoscope (GSVL) in oral and nasal intubations, no study has compared the hemodynamic responses to oral and nasal intubations with the GSVL in a single clinical trial. The purpose of this randomized clinical study was to determine whether there was a clinically relevant difference between the hemodynamic responses to oral and nasal intubations with the GSVL. METHODS: A total of 71 patients, ASA physical status I, aged 18-50 years, scheduled for elective plastic surgery under general anesthesia, were randomly allocated to the oral intubation group (OI group) and the nasal intubation group (NI group). Noninvasive blood pressure (BP) and heart rate (HR) were recorded before (baseline values) and after anesthesia induction (postinduction values), at intubation and every minute for a further 5 minutes. Maximum values of BP and HR during the observation were noted. The product of HR and systolic blood pressure (rate pressure product, RPP), the percent changes of systolic blood pressure (SBP) and HR relative to the baseline values at every measuring point, and the areas under the SBP and HR versus time curves were calculated. RESULTS: The intubation time was significantly longer in the NI group than in the OI group. The total incidence of difficulties encountered during laryngoscopy and intubation were higher in the OI group than in the NI group (29% vs. 6%, p < 0.05). BP at all measuring points, the maximum values of BP, the area under the SBP versus time curve, and the incidence of SBP percent increase more than 30% of baseline value did not differ significantly between the two groups. However, HR and RPP at intubation and their maximum values during the observation, the area under the HR versus time curve, and the incidence of HR percent increase more than 30% of baseline value were significantly higher in the OI group than in the NI group (p < 0.05). As compared with the NI group, the times required to reach the maximum values of SBP and HR were significantly shorter and the times required for recovery of SBP and HR to postinduction values were significantly longer in the OI group (p < 0.05). CONCLUSION: In anesthetized adult patients, oral and nasal intubations with the GSVL can result in a similar pressor response. However, the tachycardic response to nasal intubation using a GSVL is smaller and of shorter duration than that to oral intubation using a GSVL. PMID- 18390396 TI - Evaluation of the adequacy of general anesthesia in cesarean section by auditory evoked potential index: an observational study. AB - BACKGROUND: One of the hazards of general anesthesia for cesarean section (C/S) is the risk of intraoperative awareness with or without recall of the event. We examined the relationship between auditory evoked potential index (AAI), end tidal isoflurane concentration and hemodynamic parameters to evaluate the adequacy of our routine general anesthetic technique in C/S. METHODS: This study was carried out in 70 parturients undergoing elective C/S. A standardized anesthetic technique was applied, including thiopentone and suxamethonium for induction, and O2, N2O and isoflurane for maintenance of anesthesia. AAI, heart rate, blood pressure, end-tidal isoflurane concentration and any clinical signs of inadequate depth of anesthesia such as movement, lacrimation and coughing or jerking were continuously monitored and recorded at 17 fixed time points during surgery and anesthesia. RESULTS: Mean arterial pressure and heart rate increased at intubation, but did not show a positive correlation with the AAI values during this time. The AAI subsequently changed in the same manner as the hemodynamic parameters. There were median AAI values of 43, 37 and 27 respectively during laryngoscopy, skin incision and the time of delivery. An Emax pharmacodynamic model of AAI and end-tidal isoflurane concentration showed an r2 of 0.35 (95% CI, 0.35-0.53). According to AAI values, the majority of patients (range, 48.5-61%) had a light level of anesthesia in the time from laryngoscopy up to uterine curettage and after that they experienced an adequate (surgical or deep) level of anesthesia (range, 62.3-80%). CONCLUSION: Our current general anesthetic technique appears to provide an inadequate depth of anesthesia, especially before uterine closure as a milestone. Also, we found a moderate correlation between AAI and end-tidal isoflurane concentration and, to a lesser extent, with heart rate and mean blood pressure during isoflurane-N2O anesthesia for C/S. PMID- 18390397 TI - Factors associated with vomiting in orthopedic patients receiving patient controlled epidural analgesia. AB - BACKGROUND: Although patient-controlled epidural analgesia (PCEA) can effectively relieve postoperative pain in orthopedic patients, some adverse effects are still troublesome. We conducted this study to survey the possible risk factors related to vomiting induced by PCEA. METHODS: This retrospective study was conducted to review orthopedic patients receiving postoperative PCEA. The agent for PCEA was bupivacaine prepared as a 0.1% solution with added fentanyl (1 microg/mL). Patients' characteristics including demographic data and types of surgical procedures were collected. All patients were dichotomized into vomiting and non vomiting groups and subgroup comparisons were also performed. Stepwise logistic regression analyses were conducted to determine significant factors associated with vomiting in these patients. RESULTS: There were 320 patients (111 men, 209 women) included in the analysis. No significant differences in demographic data were noted between the groups except in sex distribution. Factors related to surgery, anesthesia and PCEA were similar between groups (p > 0.05 in all). The incidence of vomiting for orthopedic patients receiving PCEA was about 9.7% (12.4% for female, 4.5% for male). After stepwise model selection, we found female sex was the only risk factor of vomiting. The odds ratio of vomiting for female gender was 3 (95% confidence interval, 1.1-8.1). General anesthesia was not associated with vomiting in these patients. CONCLUSION: Our study demonstrated the risk factor associated with vomiting for orthopedic patients receiving PCEA was female sex. Other demographic variables and factors related to surgery or anesthesia did not have an influence on vomiting. PMID- 18390398 TI - Successful cardiopulmonary resuscitation in a morbidly obese patient with airway obstruction 10 days after tracheostomy. AB - Tracheostomy is often performed in patients requiring prolonged ventilatory support. Tracheostomy tube obstruction caused either by blood clots, mucous plugs, tube malposition and tissue granulation can lead to life-threatening complications. The risk of such complications is markedly increased in morbidly obese individuals. Here we report an incident in an 81-year-old, morbidly obese, male patient who sustained airway obstruction which resulted in cardiac arrest 10 days after tracheostomy. A 17-cm 10-ng blood clot in a tracheobronchial configuration was found to cause the obstruction. It was removed and the patient recovered after resuscitation. The etiology of the obstruction, specific management, and recommendations are discussed. PMID- 18390399 TI - Protection against large-volume regurgitated fluid aspiration by the ProSeal laryngeal mask airway. AB - The ProSeal laryngeal mask airway (PLMA) is designed to protect the airway from regurgitated fluid. However, successful channeling of large volumes of regurgitated fluid by PLMA is rarely reported. This case report states that a large volume of regurgitated fluid was successfully channeled by a PLMA. The patient was a healthy 43-year-old male, properly fasted, undergoing transurethral cystolithotomy under general anesthesia with a correctly-placed PLMA. Unexpectedly, a large volume of regurgitated fluid (> 500 mL) spurted out from the PLMA esophageal drainage tube 2 hours after surgery. Careful gastric decompression was performed and airway toilet showed no evidence of aspiration of gastric contents. An endotracheal tube was then placed in lieu of the PLMA for airway maintenance. By the end of surgery, 1300 mL of fluid was drained through the gastric tube. Extubation was smooth and the patient had an uneventful recovery. Further radiological examination revealed the existence of an unrecognized vesicorectal fistula, which allowed the cystoscopic irrigation fluid to escape to the GI tract, resulting in massive gastric regurgitation. The patient underwent fistula repair a week later and was discharged without further difficulty. This is a unique clinical case report to show PLMA can be effective in preventing aspiration when massive passive regurgitation occurs. Strategies in the management of massive regurgitation during PLMA use are discussed. PMID- 18390400 TI - Penetrating injury of the palatoglossal arch associated with use of the GlideScope videolaryngoscope in a flame burn patient. AB - The GlideScope is used to facilitate exposure of the larynx in both routine and difficult airways. A 38-year-old woman with a flame burn (second/third-degree, 40% total body surface area) and inhalation injury accompanied by acute respiratory failure under mechanical ventilation support presented for wound debridement and split thickness skin grafting. After the surgery, the endotracheal tube was reinserted successfully with a GlideScope because we anticipated a problematic airway. However, direct laryngoscopy revealed the endotracheal tube had pierced through the right palatoglossal arch. Because the consulting otolaryngologist confirmed there was no active bleeding, it was decided that no further management was needed for the wound; the patient was re intubated under direct laryngoscopy. Postoperatively, she was transferred to the intensive care unit of the burn center under mechanical ventilation. This is a report on a rare complication caused by tracheal intubation associated with the use of the GlideScope. We recommend manipulation of the GlideScope should be performed with care under vigilant surveillance to minimize injury to the oropharyngeal tissues when an endotracheal tube is advanced from the mouth to the pharynx. Potential complications should be always kept in mind when the GlideScope is used for intubation. PMID- 18390401 TI - Thoracic epidural anesthesia for a polymyositis patient undergoing awake mini thoracotomy and unroofing of a huge pulmonary bulla. AB - General anesthesia with one-lung ventilation is a conventional anesthetic strategy for most chest surgery, including resection of pulmonary bullae. However, this anesthetic management may cause alveolar barotrauma, hemodynamic instability, pulmonary atelectasis and long-term ventilator dependency. Here, we report a 64-year-old female with polymyositis and bronchiolitis obliterans organizing pneumonia who was scheduled for surgical intervention for a huge pulmonary bulla over the right upper lung. Under thoracic epidural anesthesia, with the patient maintaining clear consciousness and spontaneous breathing, a mini-thoracotomy was accomplished to unroof and partially resect the bulla. There were no perioperative complications, and the patient was satisfied with the anesthetic care. Pulmonary function tests and daily physical performance also improved postoperatively. PMID- 18390402 TI - Eclampsia following cesarean section with HELLP syndrome and multiple organ failure. AB - We present a rare case of postpartum eclampsia with overt acute heart and renal failure, in the absence of any precursive signs of preeclampsia. A 41-year-old parturient underwent elective cesarean section for the delivery of twins under spinal anesthesia. Prior to the procedure, preoperative laboratory examination revealed only traceable proteinuria but she had hypertension perioperatively. Approximately 8 hours after the cesarean section, she developed seizures, followed by evident acute heart and renal failure. The diagnosis of postpartum eclampsia with HELLP (hemolysis, elevated liver enzymes and low platelets) syndrome was established and she was admitted to the surgical intensive care unit for close care. Fortunately, the patient recovered fully and was discharged 26 days later. From this illustrative example, unexplainable and sustained hypertension following cesarean section should serve as a signal to warn the health care staff concerned about the possibility of impending life-threatening postpartum eclampsia. PMID- 18390403 TI - Intrapleural misplacement of a thoracic epidural catheter in an anesthetized patient. AB - Thoracic epidural analgesia provides adequate postoperative pain relief and favorable outcomes in major operations. However, a small number of devastating complications have been reported. Here we present a case of asymptomatic but potentially life-threatening intrapleural insertion of a thoracic epidural catheter intended for postoperative analgesia. A 39-year-old male diagnosed with esophageal carcinoma was scheduled for esophageal reconstruction. After induction of general anesthesia, a thoracic epidural catheter was inserted with a paramedian approach at the T8-9 interspace, using loss of resistance to ensure correct placement. The administration of a test dose of 2% lidocaine with epinephrine was unremarkable. After right thoracotomy, the epidural catheter was found in the right pleural cavity and was instantly removed. The patient underwent the operation smoothly and was discharged 10 days later without any sequelae. We recommend practitioners estimate the depth from the skin to the epidural space by computed tomography scan before operation and perform the placement of thoracic epidural catheter while the patient is awake to avoid accidental intrapleural misplacement. PMID- 18390404 TI - Cost analysis of three anesthetic regimens under auditory evoked potential monitoring in gynecologic laparoscopic surgery. PMID- 18390405 TI - In vitro activity of tigecycline, tetracycline and fluoroquinolones against Brucella melitensis. AB - Brucellosis is a major zoonotic disease of global importance. Forty years after active drug therapy was defined, there is no optimum antibiotic therapy for brucellosis due to relatively high rates of relapse and treatment failure. The aim of this study was to assess the in vitro susceptibility of 60 Brucella melitensis isolates obtained from blood and fluids to tigecycline, tetracycline and three quinolones (ciprofloxacin, levofloxacin, moxifloxacin). The minimum inhibitory concentrations (MICs) of these antibiotics were determined by E-test method. According to the MIC90, levofloxacin (MIC90 0.094 microg/mL) was found to be the most active agent, followed by tigecycline (MIC90 0.125 microg/mL), moxifloxacin (MIC90 0.125 microg/mL), ciprofloxacin (MIC 90 0.19 microg/mL) and tetracycline (MIC90 0.25 microg/mL). The results suggest that tigecycline can be a therapeutic alternative option for the treatment of brucellosis. PMID- 18390406 TI - Retrospective analysis of 105 cases with uterine sarcoma. AB - To evaluate the role of adjuvant therapy in survival and to identify important prognostic factors in uterine sarcoma. One hundred five patients with uterine sarcoma have been retrospectively researched to evaluate the results in this tumor group. 43.8% had leiomyosarcoma, 28.6% had endometrial stromal sarcoma and 27.6% had a malign Mullarian mixed tumor while the distribution according to the histological subgroups were found to be 42.6,16.2 and 41.2% in grade I, II and III tumors respectively. 38.1% of the patients had Radiotherapy, 18.1% had chemotherapy and 12.4% had chemoradiotherapy in addition to surgery. The distant metastases rate is 30% and the local recurrence is 16.2%. All the local recurrences and 90% of the distant metastases have occurred within the first two years. The disease free survival and overall survival rates at 3rd and 5th years are 54.46, 49.88, 54.63 and 51.09% all respectively. In our series, univariate analysis for overall survival demonstrated statistical significance for radical surgery, grade, stage, age, menopausal status and presence of RT in treatment modality, but; histology, number of mitosis, tumor size demonstrated no significance. Our data favors treatment for uterine sarcoma with radical surgery plus radiotherapy alone over 54 Gy or with chemotherapy. PMID- 18390407 TI - [The sodium pump could constitute a new target to combat glioblastomas]. AB - Malignant gliomas of which glioblastomas represent the ultimate grade of malignancy are characterized by dismal prognoses because malignant glioma cells present both important proliferation and neoangiogenesis processes and can actively migrate through the narrow extra-cellular spaces in the brain, often travelling relatively long distances, making them elusive targets for effective surgical management. Invasive malignant glioma cells show a decrease in their proliferation rates and a relative resistance to apoptosis (type I programmed cell death) as compared to the highly cellular centre of the tumour, and this may contribute to their resistance to conventional proapoptotic chemotherapy and radiotherapy. The multidisciplinary up to date treatment for glioblastoma patients combined maximal surgical removal of the tumor with postoperative radiotherapy and concomitant chemotherapy with temozolomide. Temozolomide is a proautophagic (type II programmed cell death) drug and can thus circumvent part of the glioblastoma resistance to apoptosis. Another way to potentially overcome apoptosis resistance is to decrease the migration of malignant glioma cells in the brain, which then should restore a level of sensitivity to proapoptotic drugs. The Na(+)/K(+)-ATPase or sodium pump is an ion transporter which in addition to exchanging cations, is also the ligand for cardenolides and is directly involved in the migration of cancer cells in general and of glioma cells in particular. We have shown that the alpha1 subunit of the sodium pump is highly expressed in glioma cells compared to normal brain tissues and we are the first to propose the alpha1 subunit of the sodium pump as a new target in the context of malignant glioma treatment. Using a novel cardenolide with unique structural features, which markedly inhibits sodium pump activity and binds to the alpha1 subunit, we have shown marked anti-proliferative and anti-migratory effects on human glioblastoma cells (and other cancer cell types). We have characterized at least partially the anti-cancer mechanism of action of the novel cardenolide. It is a ligand of the alpha1 subunit of the pump which impairs the proliferation and migration of glioblastoma cells by disorganizing the actin cytoskeleton and inducing severe autophagic process in glioblastoma cells. Collectively, these data suggests that the novel cardenolide is an attractive candidate for preclinical and clinical development, at least in the area of glioblastoma. This compound should reach phase I clinical trials in the summer of 2008. PMID- 18390408 TI - Hypoxic tumor cell radiosensitization: role of the iNOS/NO pathway. AB - Hypoxia is a common feature of the tumor microenvironment and a major cause of clinical radioresistance. During the last decades, several strategies to improve tumor oxygenation were developed such as breathing high oxygen content gas under hyperbaric conditions (3 atmosphere) and improving tumor perfusion by nicotinamide, in combination with carbogen breathing and accelerated radiotherapy to counteract tumor repopulation (ARCON). Other strategies to overcome hypoxia induced radioresistance are the use of hypoxic cell radiosensitizers, which mimic oxygen and enhance thereby radiation damage (e.g. the nitroimidazoles) and bioreductive drugs, which undergo intracellular reduction to form active cytotoxic species under low oxygen tension (e.g. mitomycin C and tirapazamine). A meta-analysis of all randomized trials in which some form of hypoxic modification was performed, showed an improved local control and survival, especially in cervix and head-and-neck cancer. Nevertheless, none of the discussed strategies are used in clinical routine because of feasibility and toxicity issues. We developed an alternative strategy that takes advantage of the microenvironment of solid tumors for tumor specific radiosensitization. The inducible isoform of nitric oxide synthase (iNOS) may be induced by bacterial LPS or its derivate lipid A, is expressed by a variety of solid tumors and generates NO at high rates inside tumor cells. This local production of NO results in efficient hypoxic tumor cell radiosensitization, at non-toxic extracellular concentrations of NO. In addition, iNOS is transcriptionally upregulated by hypoxia and proinflammatory cytokines such as interferon-gamma. Hence, we proposed the pro-inflammatory tumor infiltrate as a new target for radiosensitizing strategies and identified two mechanisms: First, tumor associated immune cells (macrophages, T/NK-cells) are a source of mediators that may induce the iNOS/NO pathway inside tumor cells. Second, tumor associated macrophages can produce high levels of NO that may radiosensitize bystander tumor cells. Our ongoing research is focused on combining immunostimulatory and radiosensitizing strategies. PMID- 18390409 TI - Does ionizing radiation stimulate cancer invasion and metastasis? AB - Radiotherapy (RT) is a form of local treatment used mainly for malignant tumors. Such tumors originate from mutated stem cells. During their development they do attract a variety of host cells, coined tumor-associated host cells. Malignant tumors are characterized by uncontrolled growth, invasion and metastasis, the latter being the major cause of death of patients, even when their primary tumor is under control. RT inhibits growth. There are, however, clinical data suggesting that, under some circumstances, it may stimulate metastasis. DNA is a target of ionizing radiation (IR), though not the only one. IR produces cascades of growth factors and chemokines; it activates molecules initiating multiple signaling pathways that modulate several cellular functions. We consider cancer as a network of ecosystems, including at least the founder primary tumor, the site of metastasis and the bone marrow. As these ecosystems are in continuous communication, it is not surprising that RT of the primary tumor influences metastasis. Indeed, experiments with cells in culture and with animal tumors have shown that IR stimulates invasion and metastasis and activates pro-invasive and prometastatic cellular activities through upregulation of key molecules. At certain doses and within certain time frames, IR enhances the activities of the tumor-associated host cells that support invasion and metastasis, namely: endothelial cells building new vessels; leucocytes and macrophages causing inflammation; myofibroblasts initiating desmoplasia; osteoblasts and osteoclasts establishing bone metastasis; nerve cells producing efferent growth- and invasion promoting molecules. Techniques such as spatially fractioned radiotherapy and hadron therapy may have different effects on metastasis. Taking into consideration the dose- and time-dependency of the IR-induced tumor-associated host cell reactions, these techniques, as well as the conventional ones, should be combined with repetitive biological imaging, reevaluation of planning and eventual replanning during the course of the treatment. PMID- 18390410 TI - Unraveling the biology of multiple myeloma disease: cancer stem cells, acquired intracellular changes and interactions with the surrounding micro-environment. AB - Cancerogenesis is believed to be a multistep process composed of different alterations that drive the transformation of normal human cells into highly malignant derivatives. These changes include self-sufficiency in growth signals, insensitivity to growth-inhibitory signals, the evasion of programmed cell death, a limitless replicative potential, sustained angiogenesis, and tissue invasion and metastasis. Some evidence suggests that these cancer cells are derived from immature, proliferative precursors that feed the cancer cell compartment. Both, precursors and cancer cells, intensively interact with their surroundings, commonly described as their micro-environment. The processes above are also important in the biology of multiple myeloma, a malignant bone marrow disorder consisting of monoclonal plasma cells accumulation. In the current review we describe the biology of this disease, stressing the origin of myeloma cells, their acquired genetic changes and interactions with their bone marrow microenvironment. PMID- 18390411 TI - [Cord blood: from bench to bedside]. AB - Since 1974, umbilical cord blood (CB) has been shown to contain haematopoietic stem cells similar to stem cells from the bone marrow. In 1988, E. Gluckman and her colleagues performed - successfully - the first familial CB transplantation and cured a 5 years old child suffering from Fanconi's anemia. Rapidly, CB banks were organised throughout in the world and thanks to this novel source of haematopoietic stem cells, we can now find a donor for 75 % of the patients requiring a "bone marrow" transplantation. The major benefit of CB as a source of hematopoietic stem cells is its easy access. CB also allows a more significant degree of HLA incompatibility and thus offers an opportunity of transplantation to ethnic minorities for whom no HLA identical donors are available. However, several studies have shown that the number of cells harvested in a CB was closely correlated with the engraftment post transplantation and today, a minimum of 3.7 x 10(7) mononucleated cells/kg is recommended. This required amount of cells is not always reached due to the small volume often harvested from a CB. Therefore, to apply CB transplantations to adults, different approaches are currently being investigated : coinfusion of haploidentical cells, mesenchymal cells, a second CB, or the addition of CB expanded ex-vivo. Among these approaches, double CB transplantation seems nowadays the most promising alternative and ongoing studies should soon inform us whether the duration of aplasia will be improved. PMID- 18390412 TI - Immunotherapy of hematological malignancies using dendritic cells. AB - The arsenal of therapeutic weapons against hematological malignancies is constantly growing. Unravelling the secrets of tumor immunobiology has allowed researchers to manipulate the immune system in order to stimulate tumor immunity or to bypass tumor-induced immunosuppression. An area of great interest is active specific immunotherapy where dendritic cell (DC)-based therapeutic vaccines for cancer have definitely grabbed the spotlight. DC are intensively investigated as cellular adjuvants to harness the immune system to fight off cancer by augmenting the number and effector functions of tumor-specific CD8+ cytotoxic T lymphocytes. In the present review we present a comprehensive synopsis and an update of the use of DC in hematological malignancies. In the future, more basic research as well as more clinical trials are warranted to fully establish the value of DC vaccination as an adjuvant therapy for modern hematological oncology. PMID- 18390413 TI - [General principles and first clinical trials of therapeutic vaccines against cancer]. AB - In this brief review we describe the general principles of the antigenicity of human tumor cells, which can be recognized by T lymphocytes and particularly by cytolytic T lymphocytes. This antigenicity of tumor cells lead to the development of therapeutic anticancer vaccines that should induce tumor regressions or prevent the development of metastases in the vaccinated patients. We provide some information on a set of small scale clinical trials that we have carried out with metastatic melanoma patients, and which have provided encouraging though limited clinical results. Detailed immunological analyses with some of these vaccinated patients showed strong anti-tumor T cell responses and suggested that the main limiting factor for clinical efficacy is a phenomenon of resistance of the tumor to T lymphocyte attack. Current research projects aim at elucidating the mechanisms of this resistance and to develop new vaccination strategies that circumvent this roadblock. PMID- 18390414 TI - [Has the time arrived for gene expression use at the bedside?]. AB - The microarray technology has enabled scientists to simultaneously investigate the expression of thousands of genes. Regarding breast cancer, this technology has provided a molecular classification into clinically relevant subtypes, new tools to predict disease outcome and response to treatment and new insights into carcinogenesis and metastatic progression pathways. Here we describe the state of the art of gene expression profiling for breast cancer, and we discuss the potential impact on breast cancer patient management considering its limits and promises. PMID- 18390415 TI - [HER2 and topoisomerase II alpha: useful clinical markers in breast cancer]. AB - HER-2 and potentially topoisomerase II alpha are clinically useful parameters in breast cancer. The gene status and protein expression are, for both markers, used as predictive markers in research or clinical practice. New guidelines for HER-2 testing from ASCO and CAP have been recently published and new definition for HER 2 gene amplification will probably decrease the impact of polysomy 17 on HER-2 FISH status evaluation. Different forms of HER-2 receptor (p95HER-2, p185HER-2) could be considered as prognostic or predictive markers in the future if preliminary results are confirmed. The topoisomerase II alpha case is more complex as the results of preclinical and clinical studies seem to be contradictory. The results of the clinical studies are so far, encouraging but need to be confirmed. Moreover studies should be designed to define which one of the protein or the gene or both should be evaluate as predictive markers of response to anthracyclines chemotherapy. More studies are needed today to consider this marker for daily practice. PMID- 18390416 TI - [The new generation of breast cancer clinical trials: the right drug for the right target]. AB - The principal mission of the Breast International Group (Big), Transbig, and the Breast European Adjuvant Study Team (Breast), all located at the Jules Bordet Institute in Brussels, is to accelerate and facilitate the initiation and conduct of large and difficult breast cancer clinical trials. This is made possible through the excellent network of research groups, scientists, physicians and many other collaborators deeply committed to academic clinical and translational research. A clear example of this collaboration is the HERA trial (Herceptin Adjuvant trastuzumab in HER2 positive early breast cancer) that contributed to a change of clinical practice and improved the prognosis for this particular patient population. In addition, there is an important new generation of adjuvant trials, for example, Mindact, which is evaluating the use of microarray technology in treatment decision making, and the Altto and Neo-Altto studies, which have just started and are evaluating lapatinib, a small tyrosine kinase molecule given either adjuvantly or neo-adjuvantly, alone, sequentially or in combination with trastuzumab, in patients with HER2 positive early breast cancer. The latter studies, with their strong translational research component, aim to determine which tumour profiles will best benefit from lapatinib as opposed to treatment with trastuzumab alone. PMID- 18390417 TI - [Targeted therapies in the treatment of non-small cell lung cancer]. AB - New targeted treatments offer an important opening towards the improvement of the results in the treatment of lung cancer. Currently two types of therapeutic targets are developed successfully in the treatment of advanced non-small cell lung cancer: tumour angiogenesis and growth factor receptors. Therapeutic drugs are small molecules inhibitors and the monoclonal antibodies. Bevacizumab is a monoclonal antibody against vascular endothelial growth factor receptor, VEGF, an important molecule in tumour angiogenesis. The combination of bevacizumab with first line chemotherapy increases the median survival of advanced NSCLC with a few weeks. At this moment no biomarker predicting efficacy that can help in the selection of patients is unfortunately available for this expensive treatment, although it is important to select patients based on specific contra-indications. The epidermal growth factor receptor (EGFR or HER1) is activated in NSCLC by several mechanisms. The small molecules erlotinib and gefitinib are targeting the intracellular kinase domain of the EGF receptor, thus inhibiting the signal transduction cascade. Erlotinib is currently registered and reimbursed in Belgium. The concomitant use of these small molecules with chemotherapy is ineffective in non-selected NSCLC patients. On the other hand, these molecules have great activity in patients with tumours having a constitutionally activated EGFR. Changes in the kinase domain of the receptor give rise to extremely high response rates and unexpectedly improved survival duration in patients with NSCLC. Currently studies are exploring whether erlotinib should be employed in the first line treatment of NSCLC. The FIELT study is a translational academic and multicentre phase II study in Belgium and Luxemburg addressing this issue. Specific mechanisms of resistance for these agents are gradually discovered and the new drugs being able to overcome these resistances are at the horizon. PMID- 18390418 TI - [Surgical management of colorectal liver metastases]. AB - The actual multidisciplinary treatment of patients suffering from colorectal liver metastases has benefited from major progresses both from safer and oncologically more aggressive liver resections and from more efficient chemotherapeutic agents. However, surgical resection remains still actually the only chance for potential cure and long-term survival. PMID- 18390420 TI - Image guided radiotherapy for prostate cancer. AB - The purpose of external beam radiotherapy is to sterilize malignant tumours and at the same time to avoid complications by radiation injury to the surrounding healthy tissues. Modern radiation techniques in recent years have allowed to safely escalate the dose by approximately 10% for the treatment of prostate cancer, resulting in a disease control that is nowadays comparable to surgery or permanent seed implant brachytherapy. Two recent technical developments have dramatically increased the precision of radiation dose delivery: conformal radiotherapy and image guided radiotherapy (IGRT). Conformal radiotherapy aims to shape the dose distribution to the shape of the target. At least equally important as conformality is the accurate spatial delivery of the conformal dose distribution to the target. Conventional patient positioning by skin drawings and lasers is an imprecise way to target the prostate within the pelvis. The need for adequate patient/target setup led in recent years to the development of a variety of solutions. They bear in common that setup is no longer guided by skin marks but by some imaging modality. An ideal IGRT system would allow for daily prostate imaging without possible introduction of errors due to image-acquisition itself, do so within a reasonable time frame, without the necessity for implanted radio opaque markers and preferentially without exposing the patient to radiation. A solution that combines all these features is inexistent so far. PMID- 18390419 TI - [Pancreatic cancer: ten years of systemic therapy]. AB - By today, systemic therapy for pancreatic cancer remains a huge challenge considering the last 10 years existing data. In 1997, gemcitabine emerged as a new reference treatment in advanced disease stage displaying a valuable clinical benefit rate but an overall survival limited to 6 months. In 2007, around 20 large phase III trials enrolling 7000 patients have evaluated a lot of new cytotoxic and biological compounds, but only a modest and poorly clinically relevant benefit in survival of 0.5 month was obtained, a result in sharp contrast to what it is observed in other types of cancer, where some substantial progress was noted over the last ten years. Considering the aggressive and multiresistant phenotype of this cancer, a different and more selective approach for development of new therapies has to be envisaged, starting with the deciphering of the biomolecular mechanism, first step towards the discovery of new therapeutic targets. By this way only, we could anticipate relevant advances and better exploitation and exploration of future targeted therapies. PMID- 18390421 TI - [Supportive care in cancer: concepts, achievements and challenges]. AB - The aim of supportive care in oncology is to treat the cancer related symptoms and to deal with the side effects of the treatments of the neoplastic disease. The goal of this article is to present a review of the current state of knowledge in this field by successively exposing the achievements of the last few years, the not yet solved problems and the challenges caused by the new therapeutics against cancer. This article will expose the achievements in the control of cancer related symptoms like cerebral metastases, compressive syndromes, denutrition, dyspnea, bone metastases, thromboembolic events and pain. The recent progress in the management of the side effects of chemotherapy were accomplished in treatment or prevention of mucositis, nausea, febrile neutropenia, anemia and cardiotoxicity of the anthracyclines. The unsolved problems in supportive care are alopecia, thrombocytopenia, cancer-related fatigue and cachexia. Finally, these last years saw the advent of many agents of molecular-targeted therapy in medical oncology which currently form part of the current clinical practice. These treatments have their own side effects, different from those of the cytotoxic, hormonal or immunotherapeutic agents. It is necessary to know these side effects and their management in order to provide the best quality of care to the patients who receive these treatments. PMID- 18390422 TI - [Considerations for normalisation of RT-qPCR in oncology]. AB - Gene expression analysis has many applications in the management of cancer, including diagnosis, prognosis, and therapeutic care. In this context, the reverse transcription quantitative polymerase chain reaction (RT-qPCR) has become the "gold standard" for mRNA quantification. However, this technique involves several critical steps such as RNA extraction, cDNA synthesis, quantitative PCR, and analysis, which all can be source of variation. To obtain biologically meaningful results, data normalisation is required to correct sample-to-sample variations that may be introduced during this multistage process. Normalisation can be carried out against a housekeeping gene, total RNA mass, or cell number. Careful choice of the normalization method is crucial, as any variation in the reference will introduce errors in the quantification of mRNA transcripts. By reviewing the different methods available and their related problems, the aim of this article is to provide recommendations for the set up of an appropriate normalisation strategy for RT-qPCR data in oncology. PMID- 18390423 TI - [Cellular energetic metabolism of cerebral tissue: metabolic characteristics of glial tumours]. AB - This review reports recent observations concerning specificities of the cellular energy metabolism in cerebral tissues that highlight on characteristics of that of glial tumours, such as the association of metabolic alterations aggressiveness of these tumours. Compared to normal cerebral tissue, glial tissue exhibits both a relative independence towards oxygen and substrate furnitures and thus vascularization, as well as the metabolic co-operation of neurons and glial cells within the tumour. Occurrence of a Warburg effect could explain such metabolic autonomy that might be associated to genetic changes observed in gliomas. Characteristics of the glycolytic metabolism within glioma tissue therefore may be novel land therapeutic approaches for the treatment of these tumours. PMID- 18390424 TI - [Biochemical exploration of energetic metabolism and oxidative stress in low grade gliomas: central and peripheral tumor tissue analysis]. AB - Gliomas represent 50% of primary brain tumors, and their prognosis remains poor despite the advances in diagnosis and therapeutic strategies. Low grade gliomas (LGG) are infiltrative tumors and they constantly undergo malignant transformation. Metabolic exploration of human gliomas in vivo, in animals and by using cell culture models showed important differences between tumor tissues and normal brain tissues, which can provide new markers for diagnosis, prognosis and therapeutic targets. In this study, energetic and oxidant metabolisms were explored in biopsy extracts of LGG obtained from the centre and the periphery of tumors. Metabolic pattern of these tumors was explored and the differences between the centre and the periphery pointed. Our study showed a metabolic heterogeneity between tumors, with hypermetabolic and hypometabolic profiles. Lactate to pyruvate ratio was>1, suggesting that the energy metabolism in LGG is glycolytic in nature, particularly in the centre of the tumors. Peripheral samples of tumors showed increased glucose consumption and cytochrome c oxidase activity. Lipid peroxidation and catalase activity were also increased in the periphery compared to the centre of tumors. A relationship between the main antioxidant and energy metabolism enzymes activities was observed, suggesting that periphery of tumors is more active metabolically and more resistant to free radical injury. PMID- 18390425 TI - [Oxidant stress and cardiovascular absolute risk in Tunisian type 2 diabetes]. AB - INTRODUCTION: cardiovascular diseases constitute the most hefty complications in diabetes. Absolute cardiovascular risk (ACVR) can be estimated by many equations that are continuously criticised. The aim is, in one hand, to evaluate ACVR in type 2 diabetes and, in the other hand, is to establish correlations between ACVR and oxidant-antioxidant status. MATERIAL AND METHODS: 183 type 2 diabetes and 200 controls were admitted. ACVR assessment was calculated following Laurier equation. Oxidant status was evaluated by the measure of homocysteine, hydrogen peroxide (H2O2) and LDL thiobarbituric reactive oxygen substances (LDL-TBARS). Antioxidant status was evaluated by the measure of superoxide dismutase (SOD) activity, glutathione peroxidase (GPx), total antioxidant status (TAS) vitamins A, E and zinc. Glycated haemoglobin (HbA1c) and microalbuminuria were assessed by turbidimetry. RESULTS: ninety percent of diabetes belonged to moderate and high ACVR groups. In diabetic men ACVR was doubled each elevation of 4 micromol/L homocysteine, of 50 micromol/L of H2O2 and of 20 mg/L of microalbuminuria. High risk ACVR group showed the lowest SOD activity, zincemia and the highest HbA1c. No significant difference was found in LDL-TBARS, TAS, GPx, vitamins A, E between the different ACVR groups. The strong relation between homocysteine and ACVR confirms homocysteine atherosclerotic role. Homocysteine auto-oxidation produces H2O2 leading to LDL-TBARS increase. Microalbuminuria-ACVR association verifies its vasculopathy predictor role. Urinary albumin leakage may be consequent to the hyperhomocysteinemia found in diabetes. CONCLUSION: homocysteine introduction in ACVR assessment equation may ameliorate this estimation. PMID- 18390426 TI - [Comparison and relevance of rheumatoid factors, antikeratin antibodies and anti cyclic citrullinated peptides antibodies in rheumatoid arthritis]. AB - OBJECTIVES: to evaluate specificity and sensibility of the rheumatoid factors (RF), the anti-cyclic citrullinated peptide antibodies (CCP) and the anti-keratin antibodies (AKA) according to the rheumatoid arthritis (RA) diagnosis; pathology other than RA with at least one of these marker positive; the significance of the flocculent fluorescence of the antibodies AKA by indirect immunofluorescence (IIF). METHOD: two hundred forty height patients were studied: 121 RA, 89 inflammatory rheumatisms, 23 non inflammatory rheumatisms, and 15 non rheumatic affections. The RF was investigated by nephelometry, the anti-CCP by immunofluorometry and the AKA by IIF on rat oesophagus. RESULTS: specificity and sensibility were respectively in a retrospective manner: 68% and 83% for the RF, 95% and 76% for the anti- CCP, 83% and 40% for the AKA during RA with evolution of less than one year. The rates of agreements were: RF versus CCP: 81%, RF versus AKA: 57%, CCP versus AKA: 73%. Twelve patients with pathologies different from RA have positive anti-CCP or AKA. Thirty three of the patients with anti-CCP level superior to 130 U/mL have flocculent AKA versus only 5% when the anti-CCP are lower than 130 U/mL. CONCLUSION: the RF and the anti-CCP are complementary in RA. Autoimmune and neoplasic pathologies are sometimes responsible for the positivity of the anti-CCP and the AKA. The flocculent aspect of AKA in IIF may be associated with raised concentrations of anti-CCP. PMID- 18390427 TI - [Haemolysis and turbidity influence on three analysis methods of quantitative determination of total and conjugated bilirubin on ADVIA 1650]. AB - Plasma bilirubin testing is crucial to prevent the occurrence of neonatal kernicterus. Haemolysis may occur during sampling and interfere with bilirubin determination. Moreover, lipidic infusions may induce plasma lipemia and also interfere with bilirubin measurement. We evaluated the interference of haemolysis and lipemia with three methods of total and direct bilirubin measurement adaptated on an Advia 1650 analyser (Siemens Medical Solutions Diagnostics) : Synermed (Sofibel), Bilirubin 2 (Siemens) and Bilirubin Auto FS (Diasys). The measurement of total bilirubin was little affected by haemolysis with all three methods. The Bilirubin 2 (Siemens) method was the less sensitive to haemolysis even at low bilirubin levels. The measurement of conjugated bilirubin was significantly altered by low heamoglobin concentrations for Bilirubin Auto FS(R) (30 microM or 0,192 g/100 mL haemoglobin) and for Synermed (60 microM or 0,484 g/100 mL haemoglobin). In marked contrast, we found no haemoglobin interference with the Direct Bilirubin 2 reagent which complied with the method validation criteria from the French Society for Biological Chemistry. The lipemia up to 2 g/L of Ivelip did not affect neither the measurement of total bilirubin for all three methods nor the measurement of conjugated bilirubin with the Diasys and Siemens reagents. However, we observed a strong interference starting at 0,5 g/L of Ivelip with the Synermed reagent. Our data suggest that both Siemens and Diasys methods allow to measure accurately total and conjugated bilirubin in hemolytic and lipemic samples, nevertheless, the Siemens methodology is less affected by these interferences. PMID- 18390428 TI - [Methods for microbiological control of textiles]. AB - Our laboratory is requested more and more by the establishments of health and the industrial laundries for the microbiological realization of control on the textile articles after completion. We checked the effectiveness of the techniques of the bacteriol prints or rodac and the bacterial extraction after maceration for the realization of these controls. The output of extraction of the bacteriol prints applied to woven material samples sterilized and then artificially contaminated by a titrated Staphylococcal suspension is lower than 1%. The technique recommended and largely used for the study in particular of the contamination of the blouses of doctor in the establishments of health thus does not appear to be relevant. We propose for the quality control of the linen a technique of maceration then extraction by the ultra sounds whose output is evaluated to 62% which requires to sacrifice the controled textile article. The choice of the bacteriological techniques as controls ultimate of a procedure must be carried out carefully to meet the needs for quality. Conclusions, when with the effectiveness of a process, deduced from results obtained by a bad method could be wrongfully reassuring. PMID- 18390429 TI - [Diabetic ketoacidosis with a fatal issue: is it a MODY3 (maturity-onset diabetes of the young type 3)?]. AB - Ketoacidotic coma is one of the possible diabetes mellitus first symptoms. It results from complete or relative lack of insuline and is often associated with type 1 diabetes. The authors report a case of a 45-years old woman with inaugural diabetes of which atypical features have motivated the study of MODY gene (maturity-onset diabetes of the young). Gly574ser polymorphism in the HNF-1alpha gene was found, in homozygous state, and the question of the responsibility of this polymorphism in this diabete is asked. PMID- 18390430 TI - [Lactobacillus rhamnosus septicemia in a diabetic patient associated with probiotic use: a case report]. AB - BACKGROUND: probiotic agents are increasingly used as over the counter drugs for the treatment of a variety of inflammatory and infectious conditions. The rationale of their use seems to restore a friendly bacterial flora in the gut. Complications of probiotic use, although rarely reported, can occur. OBSERVATION: we describe the case of a 54-year-old diabetic woman, who developed Lactobacillus rhamnosus septicemia while she was using probiotic oral treatment. Lactobacillus is a major component of probiotic agents. Her infection resolved after amoxicilline administration. DISCUSSION: this case highlights the complication of probiotic use, and perhaps the patient's predispositions to develop such complications. PMID- 18390431 TI - [Hemophagocytic syndrome and toxoplasmic primo-infection]. AB - Hemophagocytic syndrome (HPS) is a clinical entity that combines the clinical, biological and histological symptoms. The physiopathological mechanism involves the interaction between T lymphocytes/NK cells and macrophages, at the origin of an uncontrolled activation of the macrophages. The consequence is a hemophagocytosis extending to numerous organs, preferentially bone marrow. Clinical symptoms include cytopenia, fever unresponsive to antibiotics and multiple organ dysfunctions. Infections, lymphoproliferative disorders, cancers, systemic diseases are the most prevalent triggers or etiologies of HPS. Because of its high risk of mortality, HPS constitutes a diagnostic and therapeutic urgency. The search for an aetiology, in particular by serological testing, is essential because it conditions the treatment and thus the evolution of the disease. We report here the case of a 12 years-old boy presenting a HPS secondary to a toxoplasmic primo-infection. The objective of this work is to present the step of the biological diagnosis of HPS. Moreover, this observation allows the study of a very rare clinical presentation of toxoplasmic primo-infection, in an immunocompetant patient. PMID- 18390432 TI - [Adaptation of free and total plasma carnitine determination on the Dimension HM, X-Pand model (Dade Behring)]. AB - L-carnitine is a quaternary ammonium compound that facilitates long-chain fatty acids crossing through the mitochondrial membrane allowing their beta-oxidation. Human pathologies relatives to carnitine are mainly deficiencies, with myopathy and lipids metabolism disorders. The aim of this study was to validate the free and total carnitine plasmatic level measurement using the Dimension HM, X-Pand model analyzer (Dade Behring). Free carnitine was directly determined on plasma samples, deproteinized by using ultrafiltration, whereas total carnitine was measured after alkaline hydrolysis. Within-day and between-day imprecision were < 4,00 and 9,00 % respectively. The method is linear through 250,00 micromol/L and the limit of quantification is 3,00 micromol/L. Our method was correlated to tandem mass spectrometry (r = 0,956 for free and total carnitine). Recovery of free and total carnitine from spiked plasmas was > 99 %. On board stability of cartridge (5 days) and of calibration (at least 30 days) are in agreement with a method easily adaptable in routine laboratories. PMID- 18390433 TI - [Septicaemia with Capnocytophaga sputigena in a newborn child]. AB - We describe in this article a case of septicaemia with Capnocytophaga suptigena in a premature newborn child. The newborn child exhibited fever at birth, a light inflammatory syndrome and a respiratory failure. The germ was initially identified in the blood-culture of the newborn child by conventional techniques, and then confirmed by sequencing of the ARN 16S. After investigation, it was also found in the mother's vaginal sample. Capnocytophaga sputigena carried by the mother is certainly responsible for infection of the newborn child by ascending way. After antibiotherapy, both mother and child did not present aftereffects. This is the 16th case described in the literature; Capnocytophaga sputigena carried in vaginal area is responsible for preterm labor and for septicaemia associated to respiratory failure of the newborn child. It is thus important not to neglect this germ and to estimate its sensibility to antibiotics. PMID- 18390434 TI - [Sweat testing: review of technical requirements]. AB - The sweat test, a quantitative measurement of chloride in sweat, remains a key laboratory test to support the diagnosis of cystic fibrosis. However, because of its delicate execution, sweat test result should be interpreted with biological, clinical and genetic arguments. The following guidelines which we propose, were established in order to harmonize the practices of the sweat test. They are elaborated in a consensual way by biologists from cystic fibrosis reference centers and/or from the working group "Sweat Testing" of the National College of Biochemistry Hospital praticiens, according to the current state of knowledge on the subject, the experiment of the biologists and the recommendations established in the United States and in the United Kingdom. PMID- 18390435 TI - Antiinflammatory and analgesic activity of topical administration of Siegesbeckia pubescens. AB - Several topical formulations containing methanolic extract of Siegesbeckia pubescens was investigated for antiinflammatory and analgesic activities in rat. The effects were studied using carrageenan-induced edema and formalin testing. Piroxicam gel and methyl salicylate ointment were studied as positive control for antiinflammatory and analgesic activity, respectively. The edema inhibition of the preparations containing extract at the doses of 1-5% w/w were significantly different from the control group. The antiinflammatory effect of Siegesbeckia pubescens 4-5% w/w was similar to the effect of piroxicam gel 3 h after carrageenan injection. The analgesic activity of topical preparation with more than 4% w/w was observed in the late phase. The topical analgesic activity of the extract was less than the analgesic activity of methyl salicylate ointment. The results of the present study further confirm the use of Siegesbeckia pubescens traditionally for the treatment of painful inflammatory conditions and can be useful for the treatment of local inflammation. PMID- 18390436 TI - The effect of the ratio of two acrylic polymers on the in vitro release kinetics of ketoprofen from pellets prepared by extrusion and spheronisation technique. AB - The aim of this study was to investigate the effect of physico-chemical properties of the polymers on the release profile of ketoprofen from the pellets dosage form. Ammonio Methacrylate Copolymer Type A (Eudragit RL 30 D) & Ammonio Methacrylate Copolymer Type B (Eudragit RS 30 D) were used as release rate retarding polymers. The drug containing core pellets were prepared by extrusion spheronisation technique and subsequently coated with 15% (w/w) polymer load of the combination of Eudragit RL 30 D & Eudragit RS 30 D having ratio 1:0, 4:1, 3:2, 1:1, 2:3, 1:4, 0:1 respectively. Significant differences were found among the drug release profile from different formulations. It was revealed that Eudragit RL 30 D has the effect to increase the initial drug release more significantly where as Eudragit RS 30 D has the effect to minimize the initial drug release but increase the terminal drug release more significantly. In acid media about 50% drug was released from pellets coated only with Eudragit RL 30 D where as only 5% drug was released in case of Eudragit RS 30 D but maximum 10% drug was released from pellets when coated with the combination of Eudragit RL 30 D & Eudragit RS 30 D. In buffer media, evidence of burst release was observed for the pellets coated with Eudragit RL 30 D & Eudragit RS 30 D having ratio of 1:0, 4:1, 3:2 respectively. It was also observed that drug release increases sharply as well as the release best fit to the zero order release kinetics when pellets coated with 1:1 ratio of Eudragit RL 30 D & Eudragit RS and follows Higuchi's release kinetics when ratio was 1:0 & 3:2. The results generated in this study showed that proper selection of polymeric materials based on their physico chemical properties is important in designing sustained release pellets dosage form with suitable dissolution profile. PMID- 18390437 TI - To evaluate the role of diclofenac sodium on renal parenchyma of young albino rats. AB - To study the effects of commonly used non steroidal anti-inflammatory drug diclofenac sodium on the kidneys of young albino rats after therapeutic dose. Early clinical and pharmacological studied of oral and parental forms of diclofenac sodium began shortly after the drug was discovered. However there is scanty information about the toxicities of normal dose of diclofenac in young albino rats. The present study is therefore designed to observe the gross and microscopic changes in kidney following administration of this drug in young albino rats. For this experimental study 16 albino rats were used; they were divided into two groups, each comprising of 8 animals. Group-1 received normal saline 10ml/kg body weight while group-2 received diclofenac sodium 2mg/kg body weight for two weeks. Micrometry was done on kidney tissue for proximal and distal tubular count, their diameter and the no of cells in these tubules. Highly significant changes were observed both in proximal and distal tubules of kidney. Even in therapeutic doses, Diclofenac Sodium causes damage to kidney tubules in young albino rats. PMID- 18390438 TI - Preparation and evaluation of oxytetracycline hydrochloride microbeads for delayed release. AB - Oxytetracycline HCl microbeads were prepared with sodium alginate and pectin using ionic gelation method and evaluated for morphology, flow properties, drug content and in vitro drug release study. SEM confirmed spherical structure of microbeads with rough and porous surfaces and microbeads possessed average particle size range of 639.86 to 685.74 microm. In vitro drug release study was carried out in simulated gastric fluid (SGF) for first 2 h and simulated intestinal fluid (SIF) for next 6 h. Selected formulation was coated using enteric polymer cellulose acetate phthalate to minimize burst drug release along with delayed drug release in intestinal medium. PMID- 18390439 TI - Pharmacokinetics of calcium from calcium supplements in healthy volunteers. AB - Calcium supplementation is widely used in deficiency status and as an adjuvant in the treatment of osteoporosis. The objective of this study was to compare the oral bioavailability of calcium from tablets containing calcium fumarate to that of calcium gluconate. Twelve healthy volunteers participated in the study. Single dose, two-treatment, two-sequence-crossover, randomized design test methodology was applied. The tablets were prepared by direct compression and were subjected to tests: drug content, hardness, friability, disintegration time and in vitro dissolution studies. The preparations were compared using pharmacokinetic parameters such as the area under the plasma concentration - time curve AUC((0 11)), peak plasma concentration C(max), time to reach maximum plasma concentration T(max). No statistically significant difference was observed for any of the parameters, and the 90% confidence intervals calculated for the ratio of the logarithmically transformed AUC((0-11)) values of both formulations were within the bioequivalence limit of 0.80-1.25. It can be concluded that the two tablet preparations of calcium are likely to be bioequivalent. PMID- 18390440 TI - Formulation development and optimization of Ibuprofen tablets by direct compression method. AB - Ibuprofen is widely used as a prescription and non-prescription medicine. The aim of study is to prepare Ibuprofen tablets (200mg) using direct compression technique which is now days considered a cost effective and simple method of manufacturing. It is considered as an appropriate method for hygroscopic and thermolabile substances. In order to obtain the best, optimized product, nine different formulations were developed. Diluent (X1), disintegrant (X2) and lubricant (X3) were taken as independent variables. Weight variation (Y1), thickness (Y2), length and width (Y3), hardness (Y4), friability (Y5), disintegration (Y6), dissolution (Y7) and pharmaceutical assay (Y8) were studied as response variables. The results of all nine formulations were found within the acceptable limits conforming to those given in official compendia. However, F-6 was selected as an optimized product on the basis of high dissolution (99.05%) and Assay (100.04%). The variation of weight among the tablets of F-6 was least which showed best ratio of excipients in the formulation. Optimization has proven as an effective tool in product development. This is because no clear relationship exists between the variables. PMID- 18390441 TI - Development and validation of high performance liquid chromatography method for analysis of sibutramine hydrochloride and its impurity. AB - A simple, Precise, Rapid reproducible and selective reverse phase HPLC method has been developed for the estimation of Sibutramine Hydrochloride monohydrate and its Impurity in Bulk as well as Formulation. The analyte was resolved by using Mobile phase (Sodium Dihydrogen phosphate and Acetonitrile) at the flow rate of 1.0 Ml/Min. on Isocratic HPLC system consisting of Jasco Make UV visible Detector of model UV 1575 & Jasco make HPLC pump of model PU 1580. An ODS C- 8 RP Column (4.6mm ID, 250mm L, particle size 5 Micron, at wavelength of 230 nm. PMID- 18390442 TI - Biphasic effects of atorvastatin on inflammation. AB - Statins have been shown to exert 'pleiotropic effects' independent of their cholesterol lowering actions that include anti-inflammatory properties. We show in this study that atorvastatin dependent on the way of administration may exert anti- or pro- inflammation effects. Carrageenan-induced rat paw edema and mouse air-pouch as inflammatory models were used in this study. Animals were received statins orally prior to induction of inflammation by injection of carrageenan into rat paw or the pouch. The local effect of atorvastatin was determined by injection of the drug into the pouch. Oral administration of statins reduced both the maximal edema response and neutrophils infiltration in the inflammation zone. Lovastatin had the lowest and atorvastatin had the greatest effects. Also, in the mouse air-pouch model oral treatment by atorvastatin produced a very significant (p<0.0001) reduction in carrageenan-induced pouch leukocyte recruitment and exudates production. Concurrent administration of mevalonate reversed the anti inflammatory effect of atorvastatin. However, local injection of atorvastatin into the pouch induced a dose depend and significant increase in leukocyte recruitment into the pouch that was not reversed by co-administration of mevalonate. This study shows that atorvastatin dependent on the way of administration has both pro- and anti-inflammatory properties. Contrary to anti inflammatory effects, the pro-inflammatory responses are independent of HMG CoA reductase inhibition and can be mediated directly by atorvastatin. PMID- 18390443 TI - Comparative bioavailability and in vitro in vivo correlation of two sustained release brands of theophylline: tablets and pellets. AB - To investigate the influence of dosage forms on bioavailability, a randomized single-dose crossover study under fasting conditions was conducted using two commercially available sustained release products, Quibron SR tablets and Respro SR pellets filled Capsules containing 300mg theophylline. A group of 12 healthy, male human subjects participated in this study. Serial blood samples were collected at 0, 1, 2, 3, 4, 6, 8, 10, 12 and 24 h. Theophylline was measured by high-performance liquid chromatography while absorption profiles were derived using Wagner-Nelson equation. The bioavailability of Quibron SR tablets was compared with Respro-SR pellets filled Capsules 300 mg using pharmacokinetic parameters Cmax, Tmax, AUC(0-t), and AUC(0-alpha). In addition, the 90% confidence interval (CI) for the ratio of logarithmic transformed C(max) and AUC(0-alpha) was also used to determine bioequivalence. The T/R (test/reference) ratio of Quibron SR tablets was quite close to the prescribed limits of bioequivalence i.e. 80-125%. No statistically difference was observed between the log transformed AUC(0-agr;) (P=0.971) values as well as log transformed C(max) values (P=0.854) indicating bio-availability and the extent of absorption of two brands were comparable. Moreover, the value of correlation coefficients for % in vivo absorption versus % in vitro dissolution of the two products was calculated to be 0.9533 for Respro-SR capsule and 0.9789 for Quibron-SR tablets. PMID- 18390444 TI - m-CPP induced hypolocomotion does not interfere in the assessment of memory functions in rats. AB - Central serotonergic system plays a critical role in the regulation of memory processes in rats. Evidence suggests that a dysfunction of serotonergic system contributes to various pathological conditions. Among the multiple classes of serotonin (5-Hydroxytryptamine, 5-HT) receptors described in CNS, much attention has been devoted to the role of 5-HT2C receptor family on memory functions. A number of studies have shown that 5-HT2C receptor agonists impair memory function and also decreased locomotor activity of rats. The present study was designed to investigate the effect of different doses of 5-HT2C receptor agonist metachlorophenylpiperazine (mCPP) on locomotion and cognitive behavior in rats. Groups of adult male rats were injected mCPP intraperitoneally at doses of 1, 3 and 5 mg/kg. The learning and memory of rats were assessed by water maze (WM) and passive avoidance (PA) tests. Locomotor activity of rats was monitored by open field test. mCPP decreased locomotor activity of rats as reported earlier. A negative correlation between memory function and 5-HT2C receptor stimulation was observed in WM. Furthermore the administration of mCPP dose dependently impaired memory functions and the impairment of memory induced by mCPP was greatest at the highest dose. PA test was also performed in the present study to confirm that the decreased locomotor activity exhibited by mCPP injected rats did not affect the memory assessment in WM. Irrespective of hypolocomotion induced by mCPP, drug injected rats took less time to enter the punishable compartment which confirmed that the impairment in memory functions following mCPP was not due its effect on locomotion. It is suggested that 5-HT2C receptors might be involved in memory function probably mediating a suppressive or constraining action by decreasing dopamine levels. It can be therefore concluded that 5-HT2C receptors have a negative influence on memory function, which raises the possibility of using 5 HT2C receptor antagonists in the improvement of memory functions. PMID- 18390445 TI - The use of benzodiazepines in the aged patient: clinical and pharmacological considerations. AB - Benzodiazepines are widely used to treat anxiety and insomnia in elderly patients. The interest of this prescription is discussed in this article. The discussion is based on the pharmacological properties and adverse effects of benzodiazepines in the elderly subjects. The conclusions are that benzodiazepines should be rarely prescribed in elderly people; many patients treated by benzodiazepines should be withdrawn and other therapeutic strategies than benzodiazepines should be considered to treat anxiety and insomnia in the elderly patients. PMID- 18390446 TI - Simultaneous determination of multi drug components Theophylline, Etofylline, Guaiphenesine and Ambroxol Hydrochloride by validated RP-HPLC method in liquid dosage form. AB - The RP-HPLC (reverse phase high performance liquid chromatography) method was developed and validated for simultaneous determination of Multi drug components i.e., Theophylline, Etofylline, Guaiphenesine and Ambroxol Hydrochloride in a liquid dosage form. Chromatographic separation of the four drugs was performed on a Hypersil Phenyl BDS (25cmX4.6mm, 5mm). The mobile phase constituted of triethylamine pH 3.0 buffer: methanol (85:15) v/v was delivered at the flow rate 1.5 mL/min. Detection was performed at 235 nm. The peak purity of Theophylline, Etofylline, Guaiphenesine and Ambroxol Hydrochloride were 0.99970, 0.99979, 0.99986 and 0.99949 respectively. Calibration curves were linear with correlation coefficient between 0.99995 to 0.99997 over a concentration range of 5 to 37 microg/mL for Theophylline, 19 to 140 microg/mL for Etofylline, 20 to 149 microg/mL for Guaiphenesine and 6 to 45 microg/mL for Ambroxol hydrochloride. The relative standard deviation (RSD) was found < 2.0%. The percentage recovery was found between the range of 98.6% and 100.5% at three different levels. Robustness and ruggedness were performed and result found within the RSD of 2%. All the parameters of validation were found in the acceptance range of ICH guideline. PMID- 18390447 TI - Plant crude extracts could be the solution: extracts showing in vivo antitumorigenic activity. AB - Screening active compounds from plants lead to discover new medicinal drugs which have efficient protection and treatment roles against various diseases including cancer. In our study, extracts from different plants represent seeds of: Gossypium barbadense, Ricinus communis, Sesamum indicum, Nigella sativa, Vinca rosea and Melia azedarah; fruits of: Xanthium occidental; flowers of: Atriplex nummularia; barks of: Cinnamomum zeylanicum; latex of: Ficus carica and rhizomes of: Curcuma longa and Zingiber officinale were tested in vivo using three subsequent bioassays: the BST (Brine Shrimp Toxicity bioassay), AWD (Agar well diffusion antimicrobial bioassay) and AtPDT (Agrobacterium tumefaciens Potato Disc Tumor bioassay). AWD technique omitted any extracts have antimicrobial activities while BST omitted any extract did not has physiological activity and determined the various LC(50) of each plant extract. For the first time, using a range of concentrations in the AtPDT modified protocol allowed the detection of tumor promotion caused by extract represented by A. nummularia. Using cluster analysis leads to classifying the different plant extracts activities to six groups regarding to their toxicity, antitumor activities and both of them. The extracts from edible plants represent 50% of the first and the second group which have the highest antitumor activities represented in F. caraica (group 1) and C. longa (group 2) as well as the non-edible plant extracts of Gossypium barbadense and Ricinus communis. A comparison study between the edible and herbaceous plants different extracts for their antitumor activities was performed. We recommended using the modified protocols used in this study for investigating more plants and using crude plant extracts which have antitumor activities in cancer treatment. Edible plants, which show in vivo antitumor activities, are recommended as save sources for antitumor compounds. PMID- 18390448 TI - Electrolytes and NA(+)-K(+)-ATPase: potential risk factors for the development of diabetic nephropathy. AB - Diabetic nephropathy is the leading cause of death that affects more than 40% of diabetic patients. Its metabolic derangements are frequently accompanied with electrolyte imbalances. This study was aimed to evaluate the electrolyte homeostasis during the progression of diabetic nephropathy in various stages of developing nephropathy. Patients admitted in diabetic wards of various hospitals of Karachi were selected and divided into 4 groups with 50 individuals each. Group I (healthy normotensive, non-diabetics with normal renal functions as control). Group II (diabetic patients with normal blood pressure and renal functions). Group III (diabetic hypertensive patients without renal disease). Group IV (diabetic nephropathy patients with nephropathy). Their fasting blood samples were drawn and analyzed for the estimations of intra erythrocyte and serum electrolytes and NA(+)-K(+)-ATPase activity. Group II patients showed a significant increase in intra erythrocyte sodium, serum potassium and calcium levels where as intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium and magnesium were significantly decreased as compared to control. Group III showed a significant rise in intra erythrocyte sodium levels but intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium, calcium and magnesium were significantly lowered as compared to control. Group IV revealed a significant increase in intra erythrocyte sodium and significant decrease in intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium, calcium and magnesium levels as compared to control. The results suggest the progressive trends in electrolyte abnormalities in diabetes mellitus leading to end stage renal disease along with the abnormality of their chief transport mechanism. It points towards the potentiality of electrolytes disturbances as indicators for the progression of diabetic nephropathy and also beneficial in prognosis and treatment of the disease. PMID- 18390449 TI - Opium can differently alter blood glucose, sodium and potassium in male and female rats. AB - To determine the effects of opium on serum glucose, potassium and sodium in male and female Wistar rat, opium solution (60 mg/kg) injected intraperitoneally and the same volume of distilled water was used as control (7 rats in each group). Blood samples were collected at 0, 30, 60, 120, 240 and 360 minutes after injection from orbit cavity and the values of serum glucose, sodium (Na(+)) and potassium (K(+)) were measured. The data were then analyzed by the repeated measure ANOVA based on sex and case-control group. P < 0.05 considered as significant difference. Serum glucose increased significantly at 30, 60, 120 and 240 minutes after opium solution injection, in female rats compared to a control group. However, the male rats had this rise at 30, 60 and 120 minutes after opium solution injection compared to control group. While serum glucose in male rats was significantly higher than females at 30, 60 and 120 minutes, this value was higher in the female rats at 360 minutes. Therefore, serum glucose alterations following opium injection was significantly different in groups and in the sexes at different times. Sodium (Na(+)) rose at 60, 240 and 360 minutes significantly in all rats compared to control group. However, sodium alteration following opium injection was significantly different only between treated and control groups but sex-independent at all times. Potassium (K(+)) increased significantly at 60, 120, 240 and 360 minutes in male rats, compared to a control group. In female rats K(+) significantly raised at 30, 120, 240 and 360 minutes. Therefore, the alteration of K(+) in male and female rats was found time dependent and sex independent. According to our results, opium increased serum glucose in male and female rats differently, and it interferes with metabolic pathways differently on a gender dependent basis. Opium raised serum Na(+) and K(+), thus it interfere with water regulation and blood pressure via different mechanism. PMID- 18390450 TI - Clinical evaluation and monitoring of adverse effects for fixed multidose combination against single drug therapy in pulmonary tuberculosis patients. AB - To evaluate the clinical and therapeutic value of 4 and 3 drug fixed dose combinations verses single drug formulations to treat pulmonary tuberculosis patients. The occurrence of adverse effects was also monitored. A total of 293 patients having sputum positive pulmonary tuberculosis were enrolled (Male: 187 and Female: 106). Patients with renal, hepatic, diabetic, cardiac problem and pregnancy were excluded from study. Patients were randomly selected into three groups (A, B, C). Group A and B were given FDCs and group C was given single drug formulations. All patients received 4 drugs in the intensive phase and 3 drugs in the continuation phase. Group A showed the highest percentage of patients who achieved sputum conversion (98.9%). The numbers of days taken to achieve sputum conversion on average were the least for Group B (32 days). When comparing the adverse effects, the patients of Group C suffered the most, with 22 patients who vomited repeatedly, 3 complaining of itching, 2 with Jaundice and 1 dead. There was no significant difference in the efficacy among the three treatment regimens. However the side effects observed in all three groups strongly indicate that FDCs are safer for treating TB patients. There were no side effects in the continuation phase. PMID- 18390451 TI - Evaluation of antiaggressive activity of Eclipta alba in experimental animals. AB - The occurrence of aggression for prolonged periods of time could have a hazardous impact on the health and social wellbeing of the individual. Aggression can ensue due to exposure to an intimidating situation. Aggression is prominently seen when a disturbance occurs in the fine balance of neurotransmitters such as 5 hydroxytryptamine, gamma- aminobutyric acid, dopamine and their receptor subtypes. The present study investigated the ability of 100 and 200mg/kg of aqueous extract of Eclipta alba to circumvent aggression. Foot shock induced aggression and water competition test were utilized as models for screening of antiaggressive activity. Eclipta alba significantly minimized dominance (p<0.05) which is correlated to the level of aggression particularly with 200mg/kg in the water competition test. A tangible behavioral submission was observed with 100 and 200mg/kg and of Eclipta alba in the foot shock induced test. PMID- 18390452 TI - Searching for the heart of oncology nursing. PMID- 18390453 TI - A simple step to improve patient adherence with Zoladex. PMID- 18390454 TI - The shift of oncology inpatient care to outpatient care: the challenge of retaining expert oncology nurses. PMID- 18390455 TI - Basics of radiation treatment. PMID- 18390456 TI - The development of pericarditis following peripheral blood stem cell transplantation: a case report. PMID- 18390457 TI - Diagnosing gastric carcinoma. PMID- 18390458 TI - Adherence to oral endocrine therapy for breast cancer: a nursing perspective. AB - Many factors contribute to the success or failure of adjuvant therapy, but perhaps one of the most critical is treatment adherence. Patients do not adhere to prescribed therapy for a variety of reasons, and each one should be uncovered and addressed or managed accordingly to maintain therapeutic levels and give patients the best chance for positive outcomes. Nurses are well positioned to play a central role in promoting adherence by ensuring that patients understand the need for treatment and by identifying any underlying causes of nonadherence to therapy. When the issues are identified and addressed effectively, patients' chances for successful clinical outcomes are greatly improved. This article summarizes the issues surrounding adherence, paying specific attention to adjuvant endocrine therapy for breast cancer, and outlines strategies to reduce nonadherence that nurses can incorporate into clinical practice. PMID- 18390459 TI - The AIM Higher Initiative: new procedures implemented for assessment, information, and management of chemotherapy toxicities in community oncology clinics. AB - Chemotherapy-related toxicities are common and often undertreated in routine cancer care. Initiatives to improve toxicity management in practices may not always be effective. Quality improvement programs must engage multiple disciplines of the healthcare team and sustain efforts to institute and maintain procedures that address practice needs. The Assessment, Information, and Management (AIM) Higher Initiative, a quality improvement program undertaken at 15 community oncology practices, was initiated to improve the AIM of chemotherapy related toxicities in patients with cancer. AIM Higher focuses on improving five chemotherapy-related toxicities: neutropenia, anemia, depression and anxiety, diarrhea and constipation, and nausea and vomiting. Led by a nurse champion at each of the clinics, a variety of new procedures, processes, and tools were implemented to improve quality of care. Nurses and practice administrators can use the quality improvement processes to generate changes in procedures and practices. PMID- 18390460 TI - Evaluation and management of peripheral neuropathy in diabetic patients with cancer. AB - Recently, chemotherapy-induced peripheral neuropathy has received a great deal of attention. However, the interaction of diabetic neuropathy with potentially neurotoxic chemotherapy is far less understood. The incidence of type II diabetes has risen exponentially in the past two decades. In concert with the rise in type II diabetes, the number of individuals with diabetes who need chemotherapy for cancer also is expected to increase. Diabetic neuropathy and the neurotoxic effects of chemotherapy have a significant potential to cause functional disability. Diabetics may be most at risk for the effects of neurotoxic agents on peripheral nerve functioning, in addition to the other effects induced by chemotherapeutic agents. The purpose of this article is to review the evaluation, management, and clinical implications of peripheral neuropathy in patients with cancer and diabetes. PMID- 18390461 TI - Identifying the educational needs and concerns of newly diagnosed patients with breast cancer after surgery. AB - Many factors may interfere with the ability of women newly diagnosed with breast cancer to cope with treatment. Nurses should be aware of patients' needs during this critical time. The purpose of this study was to identify the educational needs and concerns of newly diagnosed patients with breast cancer after surgery. A phone interview with patients was conducted approximately one week after breast cancer surgery. A convenience sample of 200 patients was recruited from four hospitals within a large midwestern healthcare system. The subjects were interviewed by breast health specialists and navigators involved in their education and support and were asked to participate in a short interview during a surgical follow-up phone call. Study results indicate that fear of recurrence and anxiety regarding postoperative treatments accounted for more than 65% of the responses to the question "What concerns you most about your new diagnosis?" Emotional, social, and physical limitations were identified as most important in dealing with a breast cancer diagnosis. As breast health specialists and navigators, nurses can improve the quality of care for women with breast cancer and their families by providing additional support services and post-treatment information during the initial education and follow-up call. PMID- 18390462 TI - Intraperitoneal chemotherapy for women with ovarian cancer: nursing care and considerations. AB - Adjuvant treatment options for women with ovarian cancer following optimal surgical debulking traditionally have focused on IV taxane and/or platinum-based regimens. Combining intraperitoneal (IP) with IV therapy may offer a survival advantage over IV therapy alone in selected patients. The nursing care of women receiving IP chemotherapy involves unique assessment considerations, toxicity management, and patient teaching. Current IP chemotherapy administration guidelines are in various stages of development as the challenges of safe delivery to women with ovarian cancer undergo continued investigation. PMID- 18390463 TI - Extravasation of yttrium-90 ibritumomab tiuxetan: a case study. AB - Ibritumomab tiuxetan is a radiolabled monoclonal antibody that targets B-cell non Hodgkin lymphoma tumor cells. Yttrium-90 (Y-90), the radioisotope used to deliver the therapy, may act as a vesicant, potentially causing severe tissue damage if extravasation occurs. Particularly important to nurses is that Y-90 may not show signs or symptoms of extravasation until weeks or months after the IV injection. A case study of a man with B-cell non-Hodgkin lymphoma presented in this article shows the progression of the extravasation and steps taken to treat and eventually heal the extravasation site. Included is information about ibritumomab tiuxetan treatment, administration, eligibility criteria, and guideline recommendations for extravasation. PMID- 18390464 TI - An interdisciplinary consensus on managing skin reactions associated with human epidermal growth factor receptor inhibitors. AB - The use of human epidermal growth factor receptor (HER1/EGFR) inhibitors, such as erlotinib, cetuximab, and panitumumab, often is accompanied by the development of a characteristic spectrum of skin toxicities. Although these toxicities rarely are life threatening, they can cause physical and emotional distress for patients and caregivers. As a result, practitioners often withdraw the drug, potentially depriving patients of a beneficial clinical outcome. These reactions do not necessarily require any alteration in HER1/EGFR-inhibitor treatment and often are best addressed through symptomatic treatment. Although the evidence for using such therapies is limited, an interdisciplinary HER1/EGFR-inhibitor dermatologic toxicity forum was held in October 2006 to discuss the underlying mechanisms of these toxicities and evaluate commonly used therapeutic interventions. The result was a proposal for a simple, three-tiered grading system for skin toxicities related to HER1/EGFR inhibitors to be used in therapeutic decision making and as a framework for building a stepwise approach to intervention. PMID- 18390465 TI - Bioterrorism: class A agents and their potential presentations in immunocompromised patients. AB - A bioterrorism attack would be particularly challenging for medical professionals caring for patients with cancer who often have weakened immune systems. Knowledge of the class A agents and the potential variable presentations in immunocompromised patients is key to early recognition of an outbreak and prompt reporting. The purpose of this article is to present the class A agents: Bacillus anthracis (anthrax), botulinum toxin (botulism), variola virus (smallpox), Yersinia pestis (pneumonic plague), and Francisella tularensis (tularemia). The variable signs and symptoms that may be present in immunocompromised patients with cancer will be discussed with a focus on assessment and early recognition of an outbreak. The availability of vaccines and the implications for patients with cancer receiving these vaccines also will be discussed. PMID- 18390466 TI - Factors affecting the evolution of oncology nursing care. AB - Oncology nursing has evolved in response to population growth and changing demographics, changing regulatory requirements, decreasing lengths of inpatient hospital stays, and ongoing advances in cancer treatment, information, and biotechnology. Changes in societal perceptions of cancer and increased access to information have enabled patients to seek out knowledgeable and skilled oncology nurses. Nurses also play an ever-increasing role in rehabilitation as patients live longer with the effects of cancer and treatment. Significant outcomes achieved through nursing research include increased access to care and patient education; improved patient satisfaction, cost-effectiveness of health care, and treatment adherence; fewer hospital admissions; decreased lengths of stay; lower readmission rates; fewer emergency room visits; and lower healthcare costs. Oncology nurse researchers also have studied the economic burden of cancer treatment, limited employment options, and survivorship issues. The progress in professional oncology nursing parallels the progress in surgical, radiologic, biologic, medical, and genetic approaches to cancer treatment. The role and practice of the oncology nurse will continue to evolve in the coming decades as population demographics and healthcare systems change and new scientific and technologic discoveries are integrated into cancer care. PMID- 18390467 TI - Putting evidence into practice: evidence-based interventions for the prevention and management of constipation in patients with cancer. AB - Constipation is a major source of distress for patients with cancer, significantly affecting quality of life. It can be secondary to disease sequelae, side effects of treatment, or preexisting conditions. It often is unrecognized, underassessed, and ineffectively managed. Nurses play a key role in the prevention and management of constipation and need evidence-based interventions. This article summarizes the existing research evidence for constipation interventions and identifies gaps. Many of the strategies have been evaluated in nononcology populations; researchers should evaluate their effectiveness in oncology populations. PMID- 18390468 TI - Putting evidence into practice: evidence-based interventions for cancer-related dyspnea. AB - Despite the common occurrence of cancer-related dyspnea, a paucity of literature is available for review, especially research literature that reports interventions to control dyspnea. The Oncology Nursing Society's Putting Evidence Into Practice (PEP) initiative organized a team on nurses to examine the literature, rank the evidence, summarize the findings, and make recommendations for nursing practice to improve patient outcomes. Pharmacologic and nonpharmacologic agents have been used to treat dyspnea. Patients who received parenteral or oral immediate-release opioids demonstrated a benefit in the reduction of breathlessness; thus, parenteral or oral opioids are recommended for practice. Five interventions are listed in the effectiveness not established category and include extended-release morphine, midazolam plus morphine, nebulized opioids, the use of gas mixtures, and cognitive-behavioral therapy. This article critically examines the evidence, provides nurses with the best evidence for practice, and identifies gaps in the literature and opportunities for further research. PMID- 18390469 TI - Does milk cause cancer? PMID- 18390470 TI - Oncology care setting design and planning part I: Concepts for the oncology nurse that improve patient safety. PMID- 18390471 TI - Ensuring consistency among evidence-based practice interventions. PMID- 18390472 TI - Relaxation technique to ease dyspnea: a tool for oncology nurses. PMID- 18390473 TI - Activation of neurokinin-1 receptors during ozone inhalation contributes to epithelial injury and repair. AB - We investigated the importance of neurokinin (NK)-1 receptors in epithelial injury and repair and neutrophil function. Conscious Wistar rats were exposed to 1 ppm ozone or filtered air for 8 hours, followed by an 8-hour postexposure period. Before exposure, we administered either the NK-1 receptor antagonist, SR140333, or saline as a control. Ethidium homodimer was instilled into lungs as a marker of necrotic airway epithelial cells. After fixation, whole mounts of airway dissected lung lobes were immunostained for 5-bromo-2'-deoxyuridine, a marker of epithelial proliferation. Both ethidium homodimer and 5-bromo-2' deoxyuridine-positive epithelial cells were quantified in specific airway generations. Rats treated with the NK-1 receptor antagonist had significantly reduced epithelial injury and epithelial proliferation compared with control rats. Sections of terminal bronchioles showed no significant difference in the number of neutrophils in airways between groups. In addition, staining ozone exposed lung sections for active caspase 3 showed no apoptotic cells, but ethidium-positive cells colocalized with the orphan nuclear receptor, Nur77, a marker of nonapoptotic, programmed cell death mediated by the NK-1 receptor. An immortalized human airway epithelial cell line, human bronchial epithelial-1, showed no significant difference in the number of oxidant stress-positive cells during exposure to hydrogen peroxide and a range of SR140333 doses, demonstrating no antioxidant effect of the receptor antagonist. We conclude that activation of the NK-1 receptor during acute ozone inhalation contributes to epithelial injury and subsequent epithelial proliferation, a critical component of repair, but does not influence neutrophil emigration into airways. PMID- 18390474 TI - Urokinase expression by tumor suppressor protein p53: a novel role in mRNA turnover. AB - Lung carcinoma (H1299) cells deficient in p53 (p53(-/-)) express large amounts of urokinase-type plasminogen activator (uPA) protein and uPA mRNA, and exhibit slower degradation of uPA mRNA than that of p53-expressing nonmalignant Beas2B human airway epithelial cells. Expression of p53 protein in H1299 cells, upon transfection with p53 cDNA, suppressed basal as well as uPA-induced expression of uPA protein in both conditioned media and cell lysates, and decreased the level of steady-state uPA mRNA primarily due to increased uPA mRNA turnover. Inhibition of p53 expression by RNA silencing (SiRNA) in Beas2B cells enhanced basal and uPA mediated uPA protein and mRNA expression with stabilization of uPA mRNA. Purified p53 binds to the uPA mRNA 3' untranslated region (UTR) in a sequence-specific manner and endogenous uPA mRNA associates with p53 protein isolated from Beas2B cytosolic extracts. p53 binds to a 35-nucleotide uPA 3'UTR sequence and insertion of this sequence into beta-globin mRNA accelerates degradation of otherwise stable beta-globin mRNA. These observations confirm a new role for p53 as a uPA mRNA binding protein that down-regulates uPA mRNA stability and decreases cellular uPA expression. PMID- 18390476 TI - Cyclic strain-induced HSP27 phosphorylation modulates actin filaments in airway smooth muscle cells. AB - Mechanical stress (cyclic deformational strain) increases proteins of cytoskeletal and contractile domains in airway smooth muscle (ASM) cells in a manner that increases cell contractility. Here we studied the role of HSP27 in strain-induced microfilament formation and stability. Cultured ASM cells showed rapid phosphorylation of HSP27 upon cyclic strain within a few minutes that continued for 30 to 40 minutes. Such increases in HSP27 phosphorylation were abolished with SB 202190, a specific inhibitor of p38 mitogen-activated protein kinase (MAPK), but not by PD 98059 (an inhibitor of extracellular regulated kinase), GF109203X (an inhibitor of protein kinase C), or Y27632 (an inhibitor of Rho kinase). Direct activation of RhoA by GTPgammaS did not alter the level of HSP27 phosphorylation. Confocal microscopy revealed that cells pre-incubated with SB 202190, and/or Y27632 resulted in disorganization of stress fibers upon strain, unlike PD 98059 and GF 1092030X, suggesting that both p38 MAPK and Rho kinase were necessary for strain-induced microfilament formation. To determine the relationship between HSP27 and RhoA in strain-induced microfilament formation, cells were transfected with various isoforms of HSP27 and RhoA before strain. Co-expression of inactive HSP27 (3A-HSP27) with constitutively active EGF RhoA (RhoV14) caused diminution of microfilaments compared with constitutive active EGFP-RhoA (RhoV14) alone, suggesting that HSP27 is necessary for microfilament stability. Similarly, expression of phosphomimicking HSP27 (3D HSP27) was sufficient for retaining microfilament formation even when co expressed with the dominant-negative RhoA (EGFP-RhoN17). Thus, HSP27 activation is necessary for microfilament stability independently of RhoA activation. PMID- 18390475 TI - Hyaluronidase expression and activity is regulated by pro-inflammatory cytokines in human airway epithelial cells. AB - Hyaluronan (HA) is present at the apical surface of airway epithelium as a high molecular-weight polymer. Since HA depolymerization initiates a cascade of events that results in kinin generation and growth factor processing, in the present work we used primary cultures of human bronchial epithelial (HBE) cells grown at the air-liquid interface (ALI) to assess hyaluronidase (Hyal) activity by HA zymography, gene expression by quantitative real-time PCR, and localization by confocal microscopy. Because TNF-alpha and IL-1beta induce Hyals in other cells, we tested their effects on Hyals expression and activity. We found that Hyal-like activity is present in the apical and basolateral secretions from HBE cells where Hyals 1, 2, and 3 are expressed, and that IL-1beta acts synergistically with TNF alpha to increase gene expression and activity. Confocal microscopy showed that Hyals 1, 2, and 3 were localized intracellularly, while Hyal2 was also expressed at the apical pole associated with the plasma membrane, and in a soluble form on the apical secretions. Tissue sections from normal individuals and from individuals with asthma showed a Hyal distribution pattern similar to that observed on nontreated HBE cells or exposed to cytokines, respectively. In addition, increased expression and activity were observed in tracheal sections and in bronchoalveolar lavage (BAL) obtained from subjects with asthma when compared with normal lung donors and healthy volunteers. Our observations indicate that Hyal 1, 2, and 3 are expressed in airway epithelium and may operate in a coordinated fashion to depolymerize HA during inflammation associated with up-regulation of TNF-alpha and IL-1beta, such as allergen-induced asthmatic responses. PMID- 18390477 TI - The impact of the Austronesian expansion: evidence from mtDNA and Y chromosome diversity in the Admiralty Islands of Melanesia. AB - The genetic ancestry of Polynesians can be traced to both Asia and Melanesia, which presumably reflects admixture occurring between incoming Austronesians and resident non-Austronesians in Melanesia before the subsequent occupation of the greater Pacific; however, the genetic impact of the Austronesian expansion to Melanesia remains largely unknown. We therefore studied the diversity of nonrecombining Y chromosomal (NRY) and mitochondrial (mt) DNA in the Admiralty Islands, located north of mainland Papua New Guinea, and updated our previous data from Asia, Melanesia, and Polynesia with new NRY markers. The Admiralties are occupied today solely by Austronesian-speaking groups, but their human settlement history goes back 20,000 years prior to the arrival of Austronesians about 3,400 years ago. On the Admiralties, we found substantial mtDNA and NRY variation of both Austronesian and non-Austronesian origins, with higher frequencies of Asian mtDNA and Melanesian NRY haplogroups, similar to previous findings in Polynesia and perhaps as a consequence of Austronesian matrilocality. Thus, the Austronesian language replacement on the Admiralties (and elsewhere in Island Melanesia and coastal New Guinea) was accompanied by an incomplete genetic replacement that is more associated with mtDNA than with NRY diversity. These results provide further support for the "Slow Boat" model of Polynesian origins, according to which Polynesian ancestors originated from East Asia but genetically mixed with Melanesians before colonizing the Pacific. We also observed that non Austronesian groups of coastal New Guinea and Island Melanesia had significantly higher frequencies of Asian mtDNA haplogroups than of Asian NRY haplogroups, suggesting sex-biased admixture perhaps as a consequence of non-Austronesian patrilocality. We additionally found that the predominant NRY haplogroup of Asian origin in the Admiralties (O-M110) likely originated in Taiwan, thus providing the first direct Y chromosome evidence for a Taiwanese origin of the Austronesian expansion. Furthermore, we identified a NRY haplogroup (K-P79, also found on the Admiralties) in Polynesians that most likely arose in the Bismarck Archipelago, providing the first direct link between northern Island Melanesia and Polynesia. These results significantly advance our understanding of the impact of the Austronesian expansion and human history in the Pacific region. PMID- 18390478 TI - Positive selection and expression divergence following gene duplication in the sunflower CYCLOIDEA gene family. AB - Members of the CYCLOIDEA (CYC)/TEOSINTE-BRANCHED1 (TB1) group of transcription factors have been implicated in the evolution of zygomorphic (i.e., bilaterally symmetric) flowers in Antirrhinum and Lotus and the loss of branching phenotype during the domestication of maize. The composite inflorescences of sunflower (Helianthus annuus L. Asteraceae) contain both zygomorphic and actinomorphic (i.e., radially symmetric) florets (rays and disks, respectively), and the cultivated sunflower has evolved an unbranched phenotype in response to domestication from its highly branched wild progenitor; hence, genes related to CYC/TB1 are of great interest in this study system. We identified 10 members of the CYC/TB1 gene family in sunflower, which is more than found in any other species investigated to date. Phylogenetic analysis indicates that these genes occur in 3 distinct clades, consistent with previous research in other eudicot species. A combination of dating the duplication events and linkage mapping indicates that only some of the duplications were associated with polyploidization. Cosegregation between CYC-like genes and branching-related quantitative trait loci suggest a minor, if any, role for these genes in conferring differences in branching. However, the expression patterns of one gene suggest a possible role in the development of ray versus disk florets. Molecular evolutionary analyses reveal that residues in the conserved domains were the targets of positive selection following gene duplication. Taken together, these results indicate that gene duplication and functional divergence have played a major role in diversification of the sunflower CYC gene family. PMID- 18390479 TI - Nurse satisfaction and the implementation of minimum nurse staffing regulations. AB - In 1999, California passed the first legislation in the United States to establish minimum staffing levels for licensed nurses in hospitals. Implementation of the regulation began in 2004. This article examines whether nurses who work in hospitals in California have perceived improvements in their working conditions. A statewide sample survey of registered nurses is used, and the survey data are linked with regional data to learn whether changes in satisfaction are associated with the degree to which regional employers were expected to increase nurse staffing when the ratios were implemented. Nurse satisfaction improved between 2004 and 2006, particularly with the adequacy of RN staff, time for patient education, benefits, and clerical support. There was a significant increase in overall job satisfaction between 2004 and 2006. However, improvements in satisfaction with the adequacy of RN staff were not associated with the degree to which regional hospitals were expected to increase staffing. PMID- 18390480 TI - The use of long-term bioresorbable scaffolds for anterior cruciate ligament repair. AB - The absence of adequate options to restore full knee joint function through anterior cruciate ligament reconstruction prompts the need to develop new ligament replacement strategies. Recent focus within the ligament engineering field has been on the establishment of appropriate anterior cruciate ligament graft design requirements and evaluation methods. A range of biomaterials and graft constructions has been explored in an attempt to identify the optimal ligament replacement. Thorough and standardized evaluation methods are required throughout all phases of development, from initial in vitro bench screening through a large animal in vivo model. The initial positive clinical, gross pathologic, histologic, and mechanical results from a 12-month in vivo goat study demonstrate the potential of bioengineered ligament devices. PMID- 18390481 TI - Musculoskeletal desmoid tumors. AB - Desmoid tumors are benign tumors that exhibit varying degrees of local aggressiveness and diverse growth patterns. Magnetic resonance imaging remains the modality of choice for assessment of the nature and size of the soft-tissue lesion and involvement of surrounding structures. Treatment strategies include surgery, chemotherapy, hormonal therapy, and radiation therapy, either individually or in combination. Despite the benign nature of these tumors, multidisciplinary care is needed to provide combined treatment options. Chemotherapy in low doses is an excellent first-round treatment in any patient in whom contemplated local treatment may produce local morbidity and adjacent tissue injury. PMID- 18390482 TI - Treatment of periprosthetic humerus fractures associated with shoulder arthroplasty. AB - The incidence of periprosthetic humerus fracture associated with shoulder arthroplasty is approximately 0.6% to 3%. Fractures of the humerus occur most often intraoperatively and are more common during total shoulder arthroplasty than hemiarthroplasty because of difficulties in gaining access to the glenoid. Osteopenia, advanced age, female sex, and rheumatoid arthritis are medical comorbid factors that may contribute to humerus fractures and associated delayed healing and poorer function. When the humeral prosthetic component is loose or the fracture line overlaps the majority of the length of the prosthesis, revision with a long-stem implant should be considered. When the fracture overlaps the tip of the prosthesis and extends distally, open reduction and internal fixation is recommended. When the fracture is completely distal to the prosthesis and satisfactory alignment at the fracture site can be maintained with a fracture brace, then a trial of nonsurgical treatment is recommended. The primary goals of treatment are fracture union and pain relief. Loss of glenohumeral motion has limited the successful treatment of this challenging problem. PMID- 18390483 TI - Degenerative lumbar spondylolisthesis: trends in management. AB - Degenerative spondylolisthesis with spinal stenosis is common in elderly patients. When symptomatic, the resultant neurogenic claudication often leads to a diminished quality of life. A nonsurgical approach is an appropriate first step. Maximizing the chance of a solid arthrodesis improves the possibility of a successful long-term outcome. Treatment of this pathology has evolved over the past twenty years with the publication of numerous prospective randomized trials assessing the influence of fusion and instrumentation following decompression. Current prospective trials have evaluated the use of bone morphogenetic proteins as a substitute for autogenous bone graft. Recently, soft-tissue stabilization devices have been advocated as an alternative to fusion. Clinicians should critically evaluate these newer technologies and exercise caution regarding their use until controlled long-term trials are completed. PMID- 18390484 TI - Perioperative medical comorbidities in the orthopaedic patient. AB - Evaluation and management of medical comorbidities in the perioperative period can help improve surgical morbidity and mortality. Perioperative evaluation essentially is risk assessment and minimization. Patients undergoing orthopaedic treatment may benefit from temporizing measures to reduce systemic complications associated with some procedures. Patients at increased risk of cardiac ischemia should undergo risk stratification to determine possible perioperative interventions. Use of perioperative medications and/or consultation with specialists can help to address heart murmurs, bacterial endocarditis, prior stenting, heart failure, and hypertension. Patients with severe or unstable chronic obstructive pulmonary disease require the involvement of pulmonary care specialists. Renal failure can require nephrology consultation, particularly in cases of worsening renal function or urinary outflow obstruction. Hematologic considerations include bleeding and clotting. Prophylaxis should be used in patients with risk factors for peptic ulcer, as well as respiratory failure and hypotension. Nutritional status and liver disease also must be monitored and treated preoperatively. Orthopaedic diabetic patients should be placed on modified oral hypoglycemic or insulin regimens; recalcitrant cases merit consultation. Effective communication among all members of the patient's caregiving team is paramount. PMID- 18390485 TI - Congenital pseudarthrosis of the tibia. AB - Congenital pseudarthrosis of the tibia is characterized by anterolateral deformity of the tibia and shortening of the limb. Its etiology remains unclear. Although several classification systems have been proposed, none provides specific guidelines for management. Treatment remains challenging. The goal is to obtain and maintain union while minimizing deformity. The basic biologic considerations with surgical intervention include resection of the pseudarthrosis and bridging of the defect with stable fixation. Intramedullary stabilization, free vascularized fibula, and Ilizarov external fixation are among the most frequently used methods of treatment. In addition, bone morphogenetic protein recently has shown promise. Nevertheless, despite improvements in healing rates with congenital pseudarthrosis of the tibia, the potential for amputation in failed cases persists. PMID- 18390487 TI - Dead adipocytes, detected as crown-like structures, are prevalent in visceral fat depots of genetically obese mice. AB - Accumulation of visceral fat is a key phenomenon in the onset of obesity associated metabolic disorders. Macrophage infiltration induces chronic mild inflammation widely considered as a causative factor for insulin resistance and eventually diabetes. We previously showed that >90% of macrophages infiltrating the adipose tissue of obese animals and humans are arranged around dead adipocytes, forming characteristic crown-like structures (CLS). In this study we quantified CLS in visceral and subcutaneous depots from two strains of genetically obese mice, db/db and ob/ob. In both strains, CLS were prevalent in visceral compared with subcutaneous fat. Adipocyte size and CLS density exhibited a positive correlation both in visceral and in subcutaneous depots; however, the finding that adipocyte size was smallest and CLS density highest in visceral fat suggests a different susceptibility of visceral and subcutaneous adipocytes to death. Visceral fat CLS density was 3.4-fold greater in db/db than in ob/ob animals, which at the age at which our experimental strain was used are more prone to glucose metabolic disorders. PMID- 18390488 TI - Ethylene response factors in jasmonate signaling and defense response. PMID- 18390489 TI - Jasmonate signaling: toward an integrated view. PMID- 18390491 TI - Comparison of 3-dimensional obliquity and anisometric characteristics of anterior cruciate ligament graft positions using surgical navigation. AB - BACKGROUND: Surgical navigation allows continuous intraoperative monitoring of ACL graft anisometry and 3-dimensional obliquity. However, normative anisometry and obliquity measurements for different single-bundle anterior cruciate ligament graft positions are not well described. HYPOTHESIS: ACL Grafts placed in anteromedial and posterolateral bundle positions will have distinct anisometric profiles and 3-dimensional obliquities. A graft placed centrally in anterior cruciate ligament insertion sites will have different obliquity and anisometry than a conventional (single-bundle) graft extending from the tibia's posterolateral aspect to the femur's anteromedial aspect. STUDY DESIGN: Controlled laboratory study. METHODS: Five cadaveric knees were tested. A surgical navigation system was used to create 4 virtual graft positions in the anterior cruciate ligament footprint: (1) anteromedial, (2) posterolateral, (3) central, and (4) posterolateral tibia to anteromedial femur (conventional). Obliquity at various flexion angles and anisometry of each virtual graft's central fiber were determined. RESULTS: Anteromedial and posterolateral fibers are relatively parallel up to 30 degrees of flexion. At higher degrees of flexion, the anteromedial position is more oblique in the sagittal plane, while the posterolateral position is more oblique in the axial plane. The conventional single-bundle position is significantly more vertical than the central position in multiple planes throughout the range of motion. The anteromedial fiber is most isometric, while the posterolateral fiber is the least isometric at all flexion angles. There is no significant difference in the anisometry between the central or conventional positions at any flexion angle. The posterolateral, central, and conventional fibers were longest at full extension and slackened with progressive flexion. CONCLUSION: Anteromedial and posterolateral graft positions can be distinguished by sagittal and axial plane obliquity at flexion angles >30 degrees and by anisometry measurements. Conventional positioning produces a relatively vertical graft placement compared with the central position but has similar anisometry characteristics. Our data suggest that posterolateral, central, and conventional grafts should be fixed at or near full extension to avoid excessive tightening during motion. CLINICAL RELEVANCE: This study provides anisometry and 3-dimensional obliquity data for various graft positions using surgical navigation. The failure of single-bundle anterior cruciate ligament reconstruction to restore intact knee kinematics may be partly due to the relative vertical placement of conventional grafts compared with the central anterior cruciate ligament footprint position. PMID- 18390492 TI - Prevention of injuries among male soccer players: a prospective, randomized intervention study targeting players with previous injuries or reduced function. AB - BACKGROUND: This study was conducted to investigate whether the most common injuries in soccer could be prevented, and to determine if a simple questionnaire could identify players at increased risk. HYPOTHESIS: Introduction of targeted exercise programs to male soccer players with a history of previous injury or reduced function in the ankle, knee, hamstring, or groin will prevent injuries. STUDY DESIGN: Randomized controlled trial; Level of evidence, 2. METHODS: A total of 508 players representing 31 teams were included in the study. A questionnaire indicating previous injury and/or reduced function as inclusion criteria was used to divide the players into high-risk (HR) (76%) and low-risk (LR) groups. The HR players were randomized individually into an HR intervention group or HR control group. RESULTS: A total of 505 injuries were reported, sustained by 56% of the players. The total injury incidence was a mean of 3.2 (95% confidence interval [CI], 2.5-3.9) in the LR control group, 5.3 (95% CI, 4.6-6.0) in the HR control group (P = .0001 vs the LR control group), and 4.9 (95% CI, 4.3-5.6) in the HR intervention group (P = .50 vs the HR control group). For the main outcome measure, the sum of injuries to the ankle, knee, hamstring, and groin, there was also a significantly lower injury risk in the LR control group compared with the 2 other groups, but no difference between the HR intervention group and the HR control group. Compliance with the training programs in the HR intervention group was poor, with only 27.5% in the ankle group, 29.2% in the knee group, 21.1% in the hamstring group, and 19.4% in the groin defined as having carried out the minimum recommended training volume. CONCLUSION: The players with a significantly increased risk of injury were able to be identified through the use of a questionnaire, but player compliance with the training programs prescribed was low and any effect of the intervention on injury risk could not be detected. PMID- 18390494 TI - What is the placebo worth? PMID- 18390493 TI - Components of placebo effect: randomised controlled trial in patients with irritable bowel syndrome. AB - OBJECTIVE: To investigate whether placebo effects can experimentally be separated into the response to three components-assessment and observation, a therapeutic ritual (placebo treatment), and a supportive patient-practitioner relationship and then progressively combined to produce incremental clinical improvement in patients with irritable bowel syndrome. To assess the relative magnitude of these components. DESIGN: A six week single blind three arm randomised controlled trial. SETTING: Academic medical centre. PARTICIPANTS: 262 adults (76% women), mean (SD) age 39 (14), diagnosed by Rome II criteria for and with a score of > or =150 on the symptom severity scale. INTERVENTIONS: For three weeks either waiting list (observation), placebo acupuncture alone ("limited"), or placebo acupuncture with a patient-practitioner relationship augmented by warmth, attention, and confidence ("augmented"). At three weeks, half of the patients were randomly assigned to continue in their originally assigned group for an additional three weeks. MAIN OUTCOME MEASURES: Global improvement scale (range 1-7), adequate relief of symptoms, symptom severity score, and quality of life. RESULTS: At three weeks, scores on the global improvement scale were 3.8 (SD 1.0) v 4.3 (SD 1.4) v 5.0 (SD 1.3) for waiting list versus "limited" versus "augmented," respectively (P<0.001 for trend). The proportion of patients reporting adequate relief showed a similar pattern: 28% on waiting list, 44% in limited group, and 62% in augmented group (P<0.001 for trend). The same trend in response existed in symptom severity score (30 (63) v 42 (67) v 82 (89), P<0.001) and quality of life (3.6 (8.1) v 4.1 (9.4) v 9.3 (14.0), P<0.001). All pairwise comparisons between augmented and limited patient-practitioner relationship were significant: global improvement scale (P<0.001), adequate relief of symptoms (P<0.001), symptom severity score (P=0.007), quality of life (P=0.01). Results were similar at six week follow-up. CONCLUSION: Factors contributing to the placebo effect can be progressively combined in a manner resembling a graded dose escalation of component parts. Non-specific effects can produce statistically and clinically significant outcomes and the patient-practitioner relationship is the most robust component. TRIAL REGISTRATION: Clinical Trials NCT00065403. PMID- 18390495 TI - Universal RHD genotyping in fetuses. PMID- 18390496 TI - Effect of high throughput RHD typing of fetal DNA in maternal plasma on use of anti-RhD immunoglobulin in RhD negative pregnant women: prospective feasibility study. AB - OBJECTIVES: To assess the feasibility of applying a high throughput method, with an automated robotic technique, for predicting fetal RhD phenotype from fetal DNA in the plasma of RhD negative pregnant women to avoid unnecessary treatment with anti-RhD immunoglobulin. DESIGN: Prospective comparison of fetal RHD genotype determined from fetal DNA in maternal plasma with the serologically determined fetal RhD phenotype from cord blood. SETTING: Antenatal clinics and antenatal testing laboratories in the Midlands and north of England and an international blood group reference laboratory. PARTICIPANTS: Pregnant women of known gestation identified as RhD negative by an antenatal testing laboratory. Samples from 1997 women were taken at or before the 28 week antenatal visit. MAIN OUTCOME MEASURES: Detection rate of fetal RhD from maternal plasma, error rate, false positive rate, and the odds of being affected given a positive result. RESULTS: Serologically determined RhD phenotypes were obtained from 1869 cord blood samples. In 95.7% (n=1788) the correct fetal RhD phenotype was predicted by the genotyping tests. In 3.4% (n=64) results were either unobtainable or inconclusive. A false positive result was obtained in 0.8% (14 samples), probably because of unexpressed or weakly expressed fetal RHD genes. In only three samples (0.2%) were false negative results obtained. If these results had been applied as a guide to treatment, only 2% of the women would have received anti-RhD unnecessarily, compared with 38% without the genotyping. CONCLUSIONS: High throughput RHD genotyping of fetuses in all RhD negative women is feasible and would substantially reduce unnecessary administration of anti-RhD immunoglobulin to RhD negative pregnant women with an RhD negative fetus. PMID- 18390497 TI - Chemoprophylaxis in the prevention of leprosy. PMID- 18390498 TI - Effects of gender on performance in medicine. PMID- 18390499 TI - Follow-up of children who survive cancer. PMID- 18390500 TI - Report calls for shake up in management of sick notes: Incapacity, work, and benefits. PMID- 18390501 TI - Observational evidence for determining drug safety: Too high a pedestal. PMID- 18390502 TI - Helicobacter pylori test and treat versus proton pump inhibitor: Role of H pylori in dyspepsia. PMID- 18390503 TI - Doctors warn of premature move to polyclinics: Joined-up thinking? PMID- 18390504 TI - Do we neglect patients with multiple health problems?: Individual doctors do their best not to. PMID- 18390505 TI - A crisis of confidence: Vulnerability of healthcare staff. PMID- 18390506 TI - A crisis of confidence: Failing to explain basics can only undermine confidence. PMID- 18390507 TI - Trial participants need to be more representative of patients. PMID- 18390508 TI - Guide to diagnosing sexual abuse of children is published. PMID- 18390509 TI - Psychiatrist is struck off for wide ranging dishonesty and lack of insight. PMID- 18390510 TI - Wakefield tells GMC he was motivated by concern for autistic children. PMID- 18390512 TI - UN hears demands that 10% of spending on roads should go on safety measures. PMID- 18390513 TI - Academic freedom is at risk in dispute over Gardasil, lecturers say. PMID- 18390514 TI - New law puts NHS trusts at risk of charges of corporate manslaughter. PMID- 18390516 TI - Australian drug industry gives details of money spent courting doctors. PMID- 18390522 TI - An unlikely benefactor. PMID- 18390523 TI - Don't just blame the system. PMID- 18390524 TI - Are there too many female medical graduates? Yes. PMID- 18390525 TI - Are there too many female medical graduates? No. PMID- 18390526 TI - Rethinking global access to vaccines. PMID- 18390527 TI - Polymyalgia rheumatica. PMID- 18390528 TI - Prophylaxis against infective endocarditis: summary of NICE guidance. PMID- 18390529 TI - Commentary: Controversies in NICE guidance on infective endocarditis. PMID- 18390530 TI - Late hazards after repair of coarctation of the aorta. PMID- 18390535 TI - Application of the advanced radon diagnosis methods in the indoor building environment. AB - The health risk and dose calculations from exposure to indoor radon and its decay products are generally based on long-term integral measurements and standard ICRP recommendations. In this context, the results of assessments predicate more about human activities inside the building instead of a quality and an effectiveness of applied measures against the radon. The present paper is focused on a set of different measuring techniques and methods practically used for a classification of buildings regarding to the radon protection requirements. The fundamental quantitative and qualitative procedures of radon transport pathways and radon sources analysis, commonly named as radon diagnostic methods are performed in detail. PMID- 18390536 TI - Significance of QRS morphology in determining the prevalence of mechanical dyssynchrony in heart failure patients eligible for cardiac resynchronization: particular focus on patients with right bundle branch block with and without coexistent left-sided conduction defects. AB - AIMS: The aim of this study was to assess the significance of QRS morphology in determining the prevalence of mechanical dyssynchrony in heart failure (HF) patients considered eligible for cardiac resynchronization. METHODS AND RESULTS: A total of 200 consecutive HF patients (158 males, mean age 56 +/- 13.5 years) with standard indications for cardiac resynchronization therapy (CRT) were evaluated prospectively. The prevalence of an interventricular mechanical delay > or = 40 ms was lower in patients with pure right bundle branch block (RBBB) than that in those with RBBB plus left fascicular hemiblock (RBBB-LFH) and those with left bundle branch block (LBBB) (33 vs. 50 vs. 54%, P = 0.05). A maximal difference in peak myocardial systolic velocity among all 12 segments (Ts) > 100 ms was found in 63% of the patients with LBBB, whereas it was present in 31% of the patients with pure RBBB and in 42% of those with RBBB-LFH (P < 0.001). A standard deviation of Ts (Ts-SD) > 34 ms was present in 58% of the LBBB subjects, but in only 29% and 42% of the patients with pure RBBB and RBBB-LFH, respectively (P < 0.001). Intraventricular dyssynchrony, however, was not different in patients with pure RBBB and in those with RBBB-LFH in terms of maximal difference in Ts (P = 0.25) and Ts-SD (P = 0.17). CONCLUSIONS: Although LBBB was more often associated with intraventricular dyssynchrony, ECG sign of additional left ventricular (LV) conduction delay is not a helpful tool for the identification of intra-LV mechanical dyssynchrony in HF patients with RBBB who would benefit from CRT. PMID- 18390537 TI - Prolonged statin-associated reduction in neutrophil reactive oxygen species and angiotensin II type 1 receptor expression: 1-year follow-up. AB - AIMS: Our study investigated reactive oxygen species (ROS) generation and angiotensin II type 1 receptor (AT(1)-R) expression in primed polymorphonuclear leukocytes (PMNs) of dyslipidaemic subjects over prolonged statin treatment. METHODS AND RESULTS: Sixteen untreated dyslipidaemic subjects with moderately increased cardiovascular risk (National Cholesterol Education Program, Adult Treatment Panel III) were studied before and during long-term (1 year) simvastatin treatment. Neutrophils from dyslipidaemic subjects generated more ROS in comparison with cells from healthy control subjects. After 1 year of simvastatin treatment, ROS production (delta N-formyl-Met-Leu-Phe-induced generation and area under the curve) was significantly reduced. At baseline, AT1 R mRNA expression was also higher in dyslipidaemic subjects than in healthy controls and it was reduced after clinical treatment with simvastatin. In a subgroup of patients, a reduced angiotensin II-induced ROS generation was also observed upon clinical simvastatin treatment. Moreover, a direct effect of statin on the upregulated AT(1)-R expression was demonstrated in vitro in neutrophils of untreated dyslipidaemic subjects. CONCLUSION: A consistent reversion of pro inflammatory oxidative functional response and reduction of AT(1)-R expression in primed PMNs was observed in patients during long-term statin treatment. The AT1-R reduction over treatment may contribute to the normalization of dysregulated neutrophil activation which occurs in the pre-clinical phase of atherosclerosis. PMID- 18390538 TI - Description of a local cardiac adiponectin system and its deregulation in dilated cardiomyopathy. AB - AIMS: Despite recent advances in medical therapy, heart failure remains a leading cause for cardiovascular mortality, and its complex pathogenesis is incompletely understood. This study was performed to identify possible new therapeutic targets in dilated cardiomyopathy (DCM). METHODS AND RESULTS: Oligonucleotide microarray analysis was performed on endomyocardial biopsies (EMBs) from patients with early DCM (LVEDD > or = 55 mm, LVEF < or = 55%, n = 5) and control subjects (LVEDD < 55 mm, LVEF > 60%, no cardiac pathology, n = 4). Adiponectin, an adipocytokine involved in cellular metabolism, survival, and immunmodulation, was six-fold downregulated in DCM patients. Microarray data for adiponectin were confirmed by TaqMan-PCR (9.2-fold downregulation, control n= 9 vs. DCM n= 9, respectively, P < 0.05). Immunohistological analysis of EMBs showed significant downregulation of cardiac adiponectin protein expression independent of serum adiponectin (P = 0.36, ns) or serum TNFalpha concentrations (P = 0.46, ns). Neither the adiponectin receptor 1 (adipo-R1) nor adipo-R2 was deregulated in early DCM. Adiponectin mRNA and protein downregulation were confirmed in explanted hearts of patients with advanced DCM (LVEF < 25%, n= 8). In vitro, adiponectin incubation of neonatal rat ventricular myocytes led to activation of the pro-survival kinase PKB/Akt, increased eNOS-phosphorylation, and prevented stress-induced apoptosis of cardiomyocytes in an Akt-dependent manner. Moreover, inhibition of adiponectin secretion was accompanied by an increase in the expression of the cytokine and its receptors. CONCLUSION: These data indicate the existence of a local cardiac adiponectin system regulated independent of adiponectin and TNFalpha serum levels and its disturbance in cardiac pathology. The study suggests a role for adiponectin in the pathogenesis of DCM and implicates the adipocytokine as a possible future therapeutic target in DCM. PMID- 18390539 TI - Biogenesis of short intronic repeat 27-nucleotide small RNA from endothelial nitric-oxide synthase gene. AB - Endothelial nitric-oxide synthase (eNOS) is a constitutively expressed gene in endothelium that produces NO and is critical for vascular integrity. Previously, we reported that the 27-nucleotide (nt) repeat polymorphism in eNOS intron 4, a source of 27-nt small RNA, which inhibits eNOS expression, were associated with cardiovascular risk and expression of the eNOS gene. In the current study, we investigated the biogenesis of the intron 4-derived 27-nt small RNA. Using Northern blot, we showed that the eNOS-derived 27-nt short intronic repeat RNA (sir-RNA) expressed only in the eNOS expressing endothelial cells. Cells containing 10 x 27- or 5 x 27-nt repeats produced higher levels of 27nt sir-RNA and lower levels of eNOS mRNA than the cells with 4 x 27-nt repeats. The 27nt sir RNA was mostly present within the endothelial nuclei. When the splicing junctions of the 27-nt repeat containing intron 4 in the full-length eNOS cDNA vector were mutated, 27nt sir-RNA biogenesis was abolished. Suppression of Drosha or Dicer diminished the biogenesis of the 27nt sir-RNA. Our study suggests that the 27nt sir-RNA derived through eNOS pre-mRNA splicing may represent a new class of small RNA. The more eNOS is transcribed or higher number of the 27-nt repeats, the more 27nt sir-RNA is produced, which functions as a negative feedback self-regulator by specifically inhibiting the host gene eNOS expression. This novel molecular model may be responsible for quantitative differences between individuals carrying different numbers of the polymorphic repeats hence the cardiovascular risk. PMID- 18390540 TI - Brain-derived neurotrophic factor stimulates bone/cementum-related protein gene expression in cementoblasts. AB - Brain-derived neurotrophic factor (BDNF), recognized as essential in the developing nervous system, is involved in differentiation and proliferation in non-neuronal cells, such as endothelial cells, osteoblasts, and periodontal ligament cells. We have focused on the application of BDNF to the regeneration of periodontal tissue and indicated that BDNF promotes the regeneration of experimentally created periodontal defects. Cementoblasts form cementum, mineralized tissue, which is key to establishing a functional periodontium. The application of BDNF to the regeneration of periodontal tissue requires elucidation of the mechanism by which BDNF regulates the functions of cementoblasts. In this study, we examined how BDNF regulates the mRNA expression of bone/cementum-related proteins (alkaline phosphatase (ALP), osteopontin (OPN), and bone morphogenetic protein-2 (BMP-2)) in cultures of immortalized human cementoblast-like (HCEM) cells. BDNF elevated the mRNA levels of ALP, OPN, and BMP-2 in HCEM cells. Small interfering RNA (siRNA) for TRKB, a high affinity receptor of BDNF, siRNA for ELK-1, which is a downstream target of ERK1/2, and PD98059, an ERK inhibitor, obviated the increase in the mRNA levels. BDNF increased the levels of phosphorylated ERK1/2 and Elk-1, and the blocking of BDNF signaling by treatment with siRNA for TRKB and PD98059 suppressed the phosphorylation of ERK1/2 and Elk-1. Furthermore, BDNF increased the levels of phosphorylated c-Raf, which activates the ERK signaling pathway. These findings provide the first evidence that the TrkB-c-Raf-ERK1/2-Elk-1 signaling pathway is required for the BDNF-induced mRNA expression of ALP, OPN, and BMP-2 in HCEM cells. PMID- 18390541 TI - Biphasic regulation of HMG-CoA reductase expression and activity during wound healing and its functional role in the control of keratinocyte angiogenic and proliferative responses. AB - In this study, we determined the regulation and potential function of 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase (HMGR) during skin repair in mice. Upon skin injury, healthy mice exhibited a biphasic increase in HMGR expression and activity with elevated levels at days 3 and 13 post-wounding. In situ hybridization revealed wound margin keratinocytes as a cellular source of HMGR expression. In vitro experiments using cultured HaCaT keratinocytes uncovered epidermal growth factor (EGF), transforming growth factor (TGF)-alpha, and insulin as potent co-inducers of HMGR activity and vascular endothelial growth factor (VEGF) in the cells. Insulin-, but not EGF-mediated VEGF protein expression was functionally connected to co-induced HMGR activity, as simvastatin restrictively interfered only with insulin-induced translation of VEGF mRNA by inhibition of eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) phosphorylation. Functional ablation of insulin-induced sterol regulatory element binding protein (SREBP)-2 by siRNA abolished HMGR expression and insulin triggered VEGF protein release from keratinocytes. Simvastatin also blocked proliferation of cultured keratinocytes. The observed inhibitory effects of simvastatin on keratinocyte VEGF expression and proliferation could be reversed by mevalonate, the product of HMGR enzymatic activity. In accordance, simvastatin mediated inhibition of HMGR activity in acutely regenerating tissue of wounded mice was paralleled by a marked loss of VEGF protein expression and disturbances of normal proliferation processes in wound margin keratinocytes during skin repair. PMID- 18390542 TI - Partial rescue of the amelogenin null dental enamel phenotype. AB - The amelogenins are the most abundant secreted proteins in developing dental enamel. Enamel from amelogenin (Amelx) null mice is hypoplastic and disorganized, similar to that observed in X-linked forms of the human enamel defect amelogenesis imperfecta resulting from amelogenin gene mutations. Both transgenic strains that express the most abundant amelogenin (TgM180) have relatively normal enamel, but strains of mice that express a mutated amelogenin (TgP70T), which leads to amelogenesis imperfecta in humans, have heterogeneous enamel structures. When Amelx null (KO) mice were mated with transgenic mice that produce M180 (TgM180), the resultant TgM180KO offspring showed evidence of rescue in enamel thickness, mineral density, and volume in molar teeth. Rescue was not observed in the molars from the TgP70TKO mice. It was concluded that a single amelogenin protein was able to significantly rescue the KO phenotype and that one amino acid change abrogated this function during development. PMID- 18390544 TI - Structure of complex III with bound cytochrome c in reduced state and definition of a minimal core interface for electron transfer. AB - In cellular respiration, cytochrome c transfers electrons from cytochrome bc(1) complex (complex III) to cytochrome c oxidase by transiently binding to the membrane proteins. Here, we report the structure of isoform-1 cytochrome c bound to cytochrome bc(1) complex at 1.9 A resolution in reduced state. The dimer structure is asymmetric. Monovalent cytochrome c binding is correlated with conformational changes of the Rieske head domain and subunit QCR6p and with a higher number of interfacial water molecules bound to cytochrome c(1). Pronounced hydration and a "mobility mismatch" at the interface with disordered charged residues on the cytochrome c side are favorable for transient binding. Within the hydrophobic interface, a minimal core was identified by comparison with the novel structure of the complex with bound isoform-2 cytochrome c. Four core interactions encircle the heme cofactors surrounded by variable interactions. The core interface may be a feature to gain specificity for formation of the reactive complex. PMID- 18390543 TI - Deciphering the genetic bases of the structural diversity of phenolic glycolipids in strains of the Mycobacterium tuberculosis complex. AB - Phenolic glycolipids (PGL) play a major role in the virulence of mycobacteria, notably in strains of the Mycobacterium tuberculosis complex and in Mycobacterium leprae. The structure of the carbohydrate domain of these compounds is highly variable, and the genetic bases for these variations remain unknown. We demonstrated that the monoglycosylated PGL formed by Mycobacterium bovis differs from the triglycosylated PGL synthesized by M. tuberculosis (PGL-tb) because of the following two genetic defects: a frameshift mutation within the gene Rv2958c, encoding a glycosyltransferase involved in the transfer of the second rhamnosyl residue of the PGL-tb, and a deletion of a region that encompasses two genes, which encode a GDP-D-mannose 4,6-dehydratase and a GDP-4-keto-6-deoxy-D-mannose 3,5-epimerase/reductase, required for the formation of activated L-fucose. Expression of these three genes in M. bovis BCG allowed synthesis of PGL-tb in this recombinant strain. Additionally, we showed that all M. bovis, Mycobacterium microti, Mycobacterium pinnipedii, and some Mycobacterium africanum strains harbor the same frameshift mutation in their Rv2958c orthologs. Consistently, the structure of PGLs purified from M. africanum (harboring the Rv2958c mutation) and M. pinnipedii strains revealed that these compounds are monoglycosylated PGL. These findings explain the specificity of PGL-tb production by some strains of the M. tuberculosis complex and have important implications for our understanding of the evolution of this complex. PMID- 18390545 TI - Functional characterization of three G protein-coupled receptors for pigment dispersing factors in Caenorhabditis elegans. AB - Here, we report the identification, cloning, and functional characterization of three Caenorhabditis elegans G protein-coupled pigment dispersing factor (PDF) receptors, which we designated as Ce_PDFR-1a, -b, and -c. They represent three splice isoforms of the same gene (C13B9.4), which share a high degree of similarity with the Drosophila PDF receptor and are distantly related to the mammalian vasoactive intestinal peptide receptors (VPAC2) and calcitonin receptors. In a reverse pharmacological screen, three bioactive C. elegans neuropeptides, which were recently identified as the Drosophila PDF orthologues, were able to activate these receptors in a dose-dependent manner with nanomolar potency (isoforms a and b). Integrated green fluorescent protein reporter constructs reveal the expression of these PDF receptors in all body wall muscle cells and many head and tail neurons involved in the integration of environmental stimuli and the control of locomotion. Using a custom data analysis system, we demonstrate the involvement of this newly discovered neuropeptide signaling system in the regulation of locomotor behavior. Overexpression of PDF-2 phenocopies the locomotor defects of a PDF-1 null mutant, suggesting that they elicit opposite effects on locomotion through the identified PDF receptors. Our findings strengthen the hypothesis that the PDF signaling system, which imposes the circadian clock rhythm on behavior in Drosophila, has been functionally conserved throughout the protostomian evolutionary lineage. PMID- 18390546 TI - From oxygenase to sleep. PMID- 18390547 TI - Functional role of BLAP75 in BLM-topoisomerase IIIalpha-dependent holliday junction processing. AB - The BLAP75 protein combines with the BLM helicase and topoisomerase (Topo) IIIalpha to form an evolutionarily conserved complex, termed the BTB complex, that functions to regulate homologous recombination. BLAP75 binds DNA, associates with both BLM and Topo IIIalpha, and enhances the ability of the BLM-Topo IIIalpha pair to branch migrate the Holliday junction (HJ) or dissolve the double Holliday junction (dHJ) structure to yield non-crossover recombinants. Here we seek to understand the relevance of the biochemical attributes of BLAP75 in HJ processing. With the use of a series of BLAP75 protein fragments, we show that the evolutionarily conserved N-terminal third of BLAP75 mediates complex formation with BLM and Topo IIIalpha and that the DNA binding activity resides in the C-terminal third of this novel protein. Interestingly, the N-terminal third of BLAP75 is just as adept as the full-length protein in the promotion of dHJ dissolution and HJ unwinding by BLM-Topo IIIalpha. Thus, the BLAP75 DNA binding activity is dispensable for the ability of the BTB complex to process the HJ in vitro. Lastly, we show that a BLAP75 point mutant (K166A), defective in Topo IIIalpha interaction, is unable to promote dHJ dissolution and HJ unwinding by BLM-Topo IIIalpha. This result provides proof that the functional integrity of the BTB complex is contingent upon the interaction of BLAP75 with Topo IIIalpha. PMID- 18390548 TI - Molecular analysis of a 4-dimethylallyltryptophan synthase from Malbranchea aurantiaca. AB - Prenyltransferases are widely distributed in prokaryotes and eukaryotes and play critical roles in cell signaling, protein trafficking, and elaboration of complex molecules in secondary metabolism. Numerous prenylated natural products have been isolated from diverse microorganisms, including bacteria and fungi. These complex metabolites possess a wide range of biological activities, with some showing promise as medicinal agents. On the other hand, many prenylated secondary metabolites have been described as toxins such as ergot alkaloids that have potent psychotropic activity. We have characterized a new prenyltransferase isolated from genomic DNA of Malbranchea aurentiaca RRC1813. Enzyme specificity was investigated with a series of amino acid substrates revealing its function as a 4-dimethylallyltryptophan synthase. Polypeptide sequence alignment analysis showed that it groups with a new class of prenyltransferase enzymes that lack the typical (N/D)DXXD motif found in these polypeptides. MaPT activity was not dependent on a divalent cation cofactor, although it was reversibly inactivated by 5 mm EDTA. Analysis of kinetic parameters showed reduced enzyme efficiency upon simple modification of l-Trp. Moreover, d-Trp had 0.5% relative activity and functioned as a competitive inhibitor with a K(i) of 40.41 microm. Finally, Thr 105, Asp-179, Lys-189, and Lys-261 in MaPT were serially mutated, and the resulting lesions displayed low or complete loss of activity. This study provides a detailed characterization of a prenyltransferase in Malbranchea species, reveals two enzyme inhibitors, and through site-directed mutagenesis identified several key amino acid residues in catalysis, yielding new insights into this important yet understudied class of natural product biosynthetic enzymes. PMID- 18390549 TI - Structural and enzymatic analysis of MshA from Corynebacterium glutamicum: substrate-assisted catalysis. AB - The glycosyltransferase termed MshA catalyzes the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to 1-L-myo-inositol-1-phosphate in the first committed step of mycothiol biosynthesis. The structure of MshA from Corynebacterium glutamicum was determined both in the absence of substrates and in a complex with UDP and 1-L-myo-inositol-1-phosphate. MshA belongs to the GT-B structural family whose members have a two-domain structure with both domains exhibiting a Rossman-type fold. Binding of the donor sugar to the C-terminal domain produces a 97 degrees rotational reorientation of the N-terminal domain relative to the C-terminal domain, clamping down on UDP and generating the binding site for 1-L-myo-inositol-1-phosphate. The structure highlights the residues important in binding of UDP-N-acetylglucosamine and 1-L-myo-inositol-1 phosphate. Molecular models of the ternary complex suggest a mechanism in which the beta-phosphate of the substrate, UDP-N-acetylglucosamine, promotes the nucleophilic attack of the 3-hydroxyl group of 1-L-myo-inositol-1-phosphate while at the same time promoting the cleavage of the sugar nucleotide bond. PMID- 18390550 TI - LET-767 is required for the production of branched chain and long chain fatty acids in Caenorhabditis elegans. AB - LET-767 from Caenorhabditis elegans belongs to a family of short chain dehydrogenases/reductases and is homologous to 17beta-hydroxysterol dehydrogenases of type 3 and 3-ketoacyl-CoA reductases. Worms subjected to RNA interference (RNAi) of let-767 displayed multiple growth and developmental defects in the first generation and arrested in the second generation as L1 larvae. To determine the function of LET-767 in vivo, we exploited a biochemical complementation approach, in which let-767 (RNAi)-arrested larvae were rescued by feeding with compounds isolated from wild type worms. The arrest was only rescued by the addition of triacylglycerides extracted from worms but not from various natural sources, such as animal fats and plant oils. The mass spectrometric analyses showed alterations in the fatty acid content of triacylglycerides. Essential for the rescue were odd-numbered fatty acids with monomethyl branched chains. The rescue was improved when worms were additionally supplemented with long chain even-numbered fatty acids. Remarkably, let-767 completely rescued the yeast 3-ketoacyl-CoA reductase mutant (ybr159Delta). Because worm ceramides exclusively contain a monomethyl branched chain sphingoid base, we also investigated ceramides in let-767 (RNAi). Indeed, the amount of ceramides was greatly reduced, and unusual sphingoid bases were observed. Taken together, we conclude that LET-767 is a major 3-ketoacyl-CoA reductase in C. elegans required for the bulk production of monomethyl branched and long chain fatty acids, and the developmental arrest in let-767 (RNAi) worms is caused by the deficiency of the former. PMID- 18390551 TI - Function and structure of a prokaryotic formylglycine-generating enzyme. AB - Type I sulfatases require an unusual co- or post-translational modification for their activity in hydrolyzing sulfate esters. In eukaryotic sulfatases, an active site cysteine residue is oxidized to the aldehyde-containing C(alpha) formylglycine residue by the formylglycine-generating enzyme (FGE). The machinery responsible for sulfatase activation is poorly understood in prokaryotes. Here we describe the identification of a prokaryotic FGE from Mycobacterium tuberculosis. In addition, we solved the crystal structure of the Streptomyces coelicolor FGE homolog to 2.1 A resolution. The prokaryotic homolog exhibits remarkable structural similarity to human FGE, including the position of catalytic cysteine residues. Both biochemical and structural data indicate the presence of an oxidized cysteine modification in the active site that may be relevant to catalysis. In addition, we generated a mutant M. tuberculosis strain lacking FGE. Although global sulfatase activity was reduced in the mutant, a significant amount of residual sulfatase activity suggests the presence of FGE-independent sulfatases in this organism. PMID- 18390552 TI - Signaling properties of a non-metazoan Src kinase and the evolutionary history of Src negative regulation. AB - Choanoflagellates, unicellular organisms that are closely related to metazoans, possess cell adhesion and signaling proteins previously thought to be unique to animals, suggesting that these components may have played roles in the evolution of metazoan multicellularity. We have cloned, expressed, and purified the nonreceptor tyrosine kinase MbSrc1 from the choanoflagellate Monosiga brevicollis. The kinase has the same domain arrangement as mammalian Src kinases, and we find that the individual Src homology 3 (SH3), SH2, and catalytic domains have similar functions to their mammalian counterparts. In contrast to mammalian c-Src, the SH2 and catalytic domains of MbSrc1 do not appear to be functionally coupled. We cloned and expressed the M. brevicollis homolog of c-Src C-terminal kinase (MbCsk) and showed that it phosphorylates the C terminus of MbSrc1, yet this phosphorylation does not inhibit MbSrc to the same degree seen in the mammalian Src/Csk pair. Thus, Src autoinhibition likely evolved more recently within the metazoan lineage, and it may have played a role in the establishment of intercellular signaling in metazoans. PMID- 18390553 TI - Novel splice variants of rat CaV2.1 that lack much of the synaptic protein interaction site are expressed in neuroendocrine cells. AB - Voltage-gated Ca(2+) channels are responsible for the activation of the Ca(2+) influx that triggers exocytotic secretion. The synaptic protein interaction (synprint) site found in the II-III loop of Ca(V)2.1 and Ca(V)2.2 mediates a physical association with synaptic proteins that may be crucial for fast neurotransmission and axonal targeting. We report here the use of nested PCR to identify two novel splice variants of rat Ca(V)2.1 that lack much of the synprint site. Furthermore, we compare immunofluorescence data derived from antibodies directed against sequences in the Ca(V)2.1 synprint site and carboxyl terminus to show that channel variants lacking a portion of the synprint site are expressed in two types of neuroendocrine cells. Immunofluorescence data also suggest that such variants are properly targeted to neuroendocrine terminals. When expressed in a mammalian cell line, both splice variants yielded Ca(2+) currents, but the variant containing the larger of the two deletions displayed a reduced current density and a marked shift in the voltage dependence of inactivation. These results have important implications for Ca(V)2.1 function and for the mechanisms of Ca(V)2.1 targeting in neurons and neuroendocrine cells. PMID- 18390554 TI - Proton conductivity through the human TRPM7 channel and its molecular determinants. AB - TRPM7 is a divalent cation-permeable channel that is ubiquitously expressed. Recently, mouse TRPM7 has been shown to be sensitive to, and even permeable to, protons when heterologously expressed. Here we have demonstrated that human TRPM7 expressed either heterologously or endogenously also exhibits proton conductivity. The gene silencing of TRPM7 by small interfering RNA suppressed H+ currents in human cervical epithelial HeLa cells. In HEK293T cells transfected with human TRPM7, the inward proton conductance was suppressed by extracellular Mg2+ or Ca2+ with IC(50) values of 0.5 and 1.9 mm, respectively. Anomalous mole fraction behavior of H+ currents in the presence of Mg2+ or Ca2+ indicated that these divalent cations compete with protons for binding sites. Systematic mutation of negatively charged amino acid residues within the putative pore forming region of human TRPM7 into the neutral amino acid alanine was tested. E1047A resulted in non-functional channels, and D1054A abolished proton conductance, whereas E1052A and D1059A only partially reduced proton conductivity. Thus, it is concluded that Asp-1054 is an essential determinant of the proton conductivity, whereas Glu-1047 might be required for channel formation, and the remaining negatively charged amino acids in the pore region (Glu-1052 and Asp-1059) may play a facilitating role in the proton conductivity of human TRPM7. It is suggested that proton conductivity of endogenous human TRPM7 plays a role in physiologically/pathologically acidic situations. PMID- 18390555 TI - Identification of a novel chondroitin hydrolase in Caenorhabditis elegans. AB - Hyaluronidases have been postulated to be the enzyme acting at the initial step of chondroitin sulfate (CS) catabolism in vivo. Since chondroitin (Chn) but not hyaluronic acid (HA) has been detected in Caenorhabditis elegans, the nematode is a good model for elucidating the mechanism of the degradation of CS/Chn in vivo. Here we cloned the homolog of human hyaluronidase in C. elegans, T22C8.2. The Chn degrading activity in vitro was first demonstrated when it was expressed in COS-7 cells. The enzyme cleaved preferentially Chn. CS-A and CS-C were also depolymerized but to lesser extents, and HA was hardly degraded. In order of preference, the substrates ranked Chn >> CS-A > CS-C >> HA. The products of the degradation of Chn by the enzyme were characterized by anion-exchange high performance liquid chromatography and delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The structure of the major component in the digest was determined as GlcUAbeta1-3GalNAcbeta1 4GlcUAbeta1-3GalNAc, where GlcUA and GalNAc represent D-glucuronic acid and N acetyl-D-galactosamine, respectively, indicating that this enzyme is a Chn hydrolase, an endo-beta-galactosaminidase specific for Chn. Investigation of the effects of pH on the activity revealed the optimum pH of Chn hydrolase to be 6.0. Since Chn in C. elegans has been demonstrated to play critical roles in cell division, Chn hydrolase possibly regulates the function of Chn in vivo. This is the first demonstration of a Chn hydrolase in an animal. PMID- 18390556 TI - The reaction of alpha-synuclein with tyrosinase: possible implications for Parkinson disease. AB - Oxidative stress appears to be directly involved in the pathogenesis of Parkinson disease. Several different pathways have been identified for the production of oxidative stress conditions in nigral dopaminergic neurons, including a pathological accumulation of cytosolic dopamine with the subsequent production of toxic reactive oxygen species or the formation of highly reactive quinone species. On these premises, tyrosinase, a key copper enzyme known for its role in the synthesis of melanin in skin and hair, has been proposed to take part in the oxidative chemistry related to Parkinson disease. A study is herein presented of the in vitro reactivity of tyrosinase with alpha-synuclein, aimed at defining the molecular basis of their synergistic toxic effect. The results presented here indicate that, in conformity with the stringent specificity of tyrosinase, the exposed tyrosine side-chains are the reactive centers of alpha-synuclein. The reactivity of alpha-synuclein depends on whether it is free or membrane bound, and the chemical modifications on the tyrosinase-treated alpha-synuclein strongly influence its aggregation properties. On the basis of our results, we propose a cytotoxic model which includes a possible new toxic role for alpha-synuclein exacerbated by its direct chemical modification by tyrosinase. PMID- 18390557 TI - Two important changes and a reminder for authors of ASBMB publications: JBC, MCP, and JLR. PMID- 18390558 TI - Accelerated fractionation versus conventional fractionation radiation therapy for glottic cancer of T1-2N0M0 Phase III study: Japan Clinical Oncology Group study (JCOG 0701). AB - A randomized Phase III study was started in Japan to demonstrate the non inferiority of survival of accelerated fractionation radiation therapy (2.4 Gy/fr) with conventional fractionation radiation therapy (2 Gy/fr) in patients with T1-2N0M0 glottic cancer. This study began in September 2007, and a total of 360 patients will be accrued from 22 institutions within 4 years. The primary endpoint is 3-year progression-free survival (PFS). The secondary endpoints are overall survival, local progression-free survival, disease-free survival, survival with preserved voice function, complete response rate, proportion of treatment completion and adverse events. PMID- 18390559 TI - Intrauterine insemination with double ejaculate compared with single ejaculate in male factor infertility: a pilot study. AB - The aim of this study was to compare the efficiency of using a double ejaculate with the efficiency of using a single ejaculate for intrauterine insemination in male subfertility. Eligibility for the study was a total motile sperm count between 1 x 10(6) and 5 x 10(6) on postwash sperm analysis. Thirty-nine couples were randomized to the study group, while another 50 couples were randomized to serve as the control group. Males in the study group were asked to produce a second semen sample within 2 hours of the first sample on the day of insemination. In the study group, the mean total motile sperm count in the first sample was 3.83 x 10(6) (SD +/- 0.85 x 10(6); range 1.1-4.9) and 3.99 x 10(6) (SD +/- 0.72 x 10(6); range 0.9-4.4) in the control group. The mean total motile sperm count in the second sample was 3.52 x 10(6) (SD +/- 1.46 x 10(6); range 0.9 3.7) in the study group. The mean total motile sperm count in the final inseminate was 7.35 x 10(6) (SD +/- 1.90 x 10(6); range 2.9-10.6) in the study group. The difference in total motile sperm counts between the study and the control group was statistically significant (P < .001). There were 6 pregnancies in the study group, providing a pregnancy rate of 15.3%, whereas there were 5 pregnancies in the control group, representing a pregnancy rate of 10% (P = .44). We concluded that although it does not increase the pregnancy rate significantly, obtaining a second semen sample when the motile sperm yield of the first semen sample is 1 million to 5 million significantly increases the total motile sperm count in the final inseminate. PMID- 18390560 TI - Mutation screening and association study of the TSSK4 Gene in Chinese infertile men with impaired spermatogenesis. AB - The testis-specific serine/threonine kinase (TSSK) family is a specific kinase group with exclusive or dominant expression in testis and involvement in spermatogenesis and male infertility. TSSK4 is a newly identified member of the TSSK family. In order to investigate the possible relationships between variations, including mutations and polymorphisms of the TSSK4 gene and impaired spermatogenesis in humans, mutation screening of this gene in 372 patients with azoospermia or severe oligospermia and 220 controls was performed. In total, 4 novel single nucleotide changes including c.679G>A, c.987+108G>A, c.-155C>G and c.765C>A were discovered. The latter 2 variations were found only in patients, not in controls. Bioinformatics analysis suggested that allele A of c.765C>A could decrease the activity of pre-mRNA splicing of TSSK4. The frequency of allele A of c.679G>A was significantly higher in controls than in patients. On the contrary, allele A of c.987+108G>A was remarkably increased in patients compared with controls. Our investigation of TSSK4, a potentially important testicular gene, in Chinese infertile and control men identified the association of some single nucleotide polymorphisms in this gene with male infertility. PMID- 18390561 TI - A common protamine 1 promoter polymorphism (-190 C->A) correlates with abnormal sperm morphology and increased protamine P1/P2 ratio in infertile patients. AB - It is known that targeting the protamine 1 gene in mice leads to infertility, abnormal chromatin packaging, and abnormal sperm morphology. Because many infertile patients also have an abnormal sperm morphology and chromatin packaging, the human protamine 1 gene (PRM1) is an important candidate to screen for potential mutations. In this work, we have screened the PRM1 gene in search of potential mutations and determined the sperm morphology and the ratio between protamine 1 and protamine 2 (P1/P2 ratio). Direct sequencing of the PRM1 promoter led to the identification of a common single-nucleotide polymorphism (SNP; -190 C ->A). The -190 AA genotype was detected at a higher frequency (13.8%) in patients with markedly altered sperm morphology (A change was also consistently higher (.331) in infertile patients with a markedly altered morphology compared with population controls (.178; P < .01). Additionally, we have determined that the P1/P2 ratio is significantly increased in patients with the PRM1 -190 AA genotype compared with patients with the CA or CC genotypes (P = .006, Mann-Whitney). These findings indicate that the common PRM1 -190 C-->A polymorphism identified is associated with abnormal sperm head morphology and abnormal P1/P2 ratio in infertile patients. PMID- 18390562 TI - Differential effects of insular and ventromedial prefrontal cortex lesions on risky decision-making. AB - The ventromedial prefrontal cortex (vmPFC) and insular cortex are implicated in distributed neural circuitry that supports emotional decision-making. Previous studies of patients with vmPFC lesions have focused primarily on decision-making under uncertainty, when outcome probabilities are ambiguous (e.g. the Iowa Gambling Task). It remains unclear whether vmPFC is also necessary for decision making under risk, when outcome probabilities are explicit. It is not known whether the effect of insular damage is analogous to the effect of vmPFC damage, or whether these regions contribute differentially to choice behaviour. Four groups of participants were compared on the Cambridge Gamble Task, a well characterized measure of risky decision-making where outcome probabilities are presented explicitly, thus minimizing additional learning and working memory demands. Patients with focal, stable lesions to the vmPFC (n = 20) and the insular cortex (n = 13) were compared against healthy subjects (n = 41) and a group of lesion controls (n = 12) with damage predominantly affecting the dorsal and lateral frontal cortex. The vmPFC and insular cortex patients showed selective and distinctive disruptions of betting behaviour. VmPFC damage was associated with increased betting regardless of the odds of winning, consistent with a role of vmPFC in biasing healthy individuals towards conservative options under risk. In contrast, patients with insular cortex lesions failed to adjust their bets by the odds of winning, consistent with a role of the insular cortex in signalling the probability of aversive outcomes. The insular group attained a lower point score on the task and experienced more 'bankruptcies'. There were no group differences in probability judgement. These data confirm the necessary role of the vmPFC and insular regions in decision-making under risk. Poor decision making in clinical populations can arise via multiple routes, with functionally dissociable effects of vmPFC and insular cortex damage. PMID- 18390564 TI - Floristic relationships among vegetation types of new zealand and the southern andes: similarities and biogeographic implications. AB - BACKGROUND AND AIMS: Similarities between the floras of geographically comparable regions of New Zealand (NZ) and the southern Andes (SA) have interested biologists for over 150 years. The present work selects vegetation types that are physiognomically similar between the two regions, compares their floristic composition, assesses the environmental factors that characterize these matching vegetation types, and determines whether phylogenetic groups of ancestral versus modern origin are represented in different proportions in their floras, in the context of their biogeographic history. METHODS: Floristic relationships based on 369 genera of ten vegetation types present in both regions were investigated with correspondence analysis (CA) and ascending hierarchical clustering (AHC). The resulting ordination and classification were related to the environmental characteristics of the different vegetation types. The proportions of different phylogenetic groups between the regions (NZ, SA) were also compared, and between forest and non-forest communities. KEY RESULTS: Floristic similarities between NZ and SA tend to increase from forest to non-forest vegetation, and are highest in coastal vegetation and bog. The floras of NZ and SA also differ in their phylogenetic origin, NZ being characterized by an 'excess' of genera of basal origin, especially in forests. CONCLUSIONS: The relatively low similarities between forests of SA and NZ are related to the former being largely of in situ South American and Gondwanan origin, whereas the latter have been mostly reconstituted though transoceanic dispersal of propagules since the Oligocene. The greater similarities among non-forest plant communities of the two regions result from varied dispersal routes, including relatively recent transoceanic dispersal for coastal vegetation, possible dispersal via a still-vegetated Antarctica especially for bog plants, and independent immigration from Northern Hemisphere sources for many genera of alpine vegetation and grassland. PMID- 18390563 TI - Loss of ischaemic preconditioning in ovariectomized rat hearts: possible involvement of impaired protein kinase C epsilon phosphorylation. AB - AIMS: The aims of this study were to determine whether chronic oestrogen withdrawal influences the development of ischaemic preconditioning (IPC) in female hearts, to investigate the mechanism whereby IPC is impaired, and to assess whether direct activation of protein kinase C (PKC) can mimic IPC in female hearts with chronic oestrogen depletion. METHODS AND RESULTS: We performed Sham-operation (Sham) or bilateral ovariectomy on 16-week-old Sprague-Dawley female rats. Ovariectomized rats were randomized to subcutaneous implantation of 17beta-estradiol (OxE) or placebo (OxP) pellets. Four weeks later, isolated, perfused hearts were subjected to 30 min of ischaemia followed by 120 min of reperfusion with or without three cycles of 5 min ischaemia/5 min reperfusion. The cardioprotective effect of IPC was completely lost in the OxP group. Western immunoblots revealed that in the OxP group, IPC failed to translocate PKCepsilon to the membranous fraction and that phosphorylation of PKCepsilon (Ser(729)) and phosphoinositide-dependent kinase (PDK) 1 (Ser(241)) was impaired. Oestrogen replacement restored the IPC effect, the translocation and phosphorylation of PKCepsilon, and the phosphorylation of PDK1. In the OxP group, pre-treatment with a PKCepsilon selective activator peptide (Psi-epsilonRACK) mimicked the IPC effect. Pre-treatment with a phosphatidylinositol-3 kinase inhibitor before IPC abrogated the translocation and phosphorylation of PKCepsilon in the Sham group. CONCLUSIONS: The cardioprotective effect of IPC is lost in female hearts with chronic oestrogen withdrawal and this is due, at least in part, to impaired translocation and phosphorylation of PKCepsilon. Selective activation of PKCepsilon-mediated signalling can fully restore the IPC effect in a manner analogous to oestrogen replacement. PMID- 18390565 TI - No way in & no way out: a case of renal failure due to both pre- and post-renal obstruction. PMID- 18390566 TI - Body mass index, muscle and fat in chronic kidney disease: questions about survival. AB - The human body can be roughly divided into two major compartments, fat mass and lean body mass. Adipose tissue is now considered to be a highly active tissue and, in addition to storing calories as triglycerides, it also secretes a large variety of compounds, including cytokines, chemokines and hormone-like factors such as leptin, adiponectin and resistin. On the other hand, muscle plays a central role in whole-body protein metabolism by serving as the principal provider for amino acids to maintain protein synthesis in vital tissues and organs and by providing hepatic gluconeogenic precursors. Although not a good indicator of body composition, the Quetelet index, also called body mass index (BMI), is often used for practical reasons. It is well known that high BMI predicts mortality and cardiovascular disease (CVD) in the general population. However, observational reports in the dialysis population have suggested that obesity is associated with improved survival, a phenomenon that is not well understood and subject to controversies. This review describes the characteristics of BMI in the general population and in chronic kidney disease (CKD) patients, as well as the respective role of muscle, whole body fat and fat distribution towards mortality, with particular emphasis on patients with CKD. PMID- 18390567 TI - Clinical effectiveness and quality of life of conventional haemodialysis versus short daily haemodialysis: a systematic review. AB - BACKGROUND: End-stage renal disease is a troublesome health problem worldwide. The most usual renal replacement therapy is conventional haemodialysis (CHD), performed three times a week, 3.5-4 h per session. It has been proposed that this schedule is unphysiologic and that daily haemodialysis would be a more appropriate schedule. One of the variants of daily haemodialysis is the so-called short daily haemodialysis (SDHD), performed five to seven times per week, 1.5-3 h per session. The objective of this paper is to compare, through a systematic review, the clinical effectiveness and safety of SDHD versus CHD. METHODS: The following databases were searched: MEDLINE, EMBASE, NHS Centre for Reviews and Dissemination (HTA, DARE and NHS EED), Cochrane, ISI Web of Knowledge, IME and IBECS. Two independent reviewers decided which papers were to be included after applying inclusion and exclusion criteria. Any discrepancy was resolved by consensus. The quality of the included papers was measured using a quality scale developed for the purpose of this report. RESULTS: Seventeen original articles were included. There were no randomized controlled trials. SDHD seems to be more effective than conventional dialysis. Patients on daily haemodialysis seem to present less vascular access problems, better control of hypertension and in turn a reduction in the antihypertensive treatment, better quality of life, lower incidence of ventricular hypertrophy, lower consumption of rHuEPO due to the better control of anaemia and a reduction in the use of phosphate binders as a consequence of the better control of plasmatic phosphorous. CONCLUSIONS: SDHD might result in a better clinical effectiveness, mainly through a better control of the arterial tension and, therefore, a lower consumption of antihypertensive drugs, and a better quality of life than CHD. PMID- 18390568 TI - Transcriptome analysis of a cDNA library from adult human epididymis. AB - Mammalian Gene Collection (MGC) verified over 9,000 human full-ORF genes and FLJ Program reported 21,243 cDNAs of which 14,409 were unique ones and 5,416 seemed to be protein-coding. The pity is that epididymis cDNA library was missing in their sequencing target list. Epididymis is a very important male accessory sex organ for sperm maturation and storage. Fully differentiated spermatozoa left from testis acquire their motility and capacity for fertilization via interactions with the epididymal epithelium duct lumen during passage through this convoluted duct. Here, we report that 20,000 clones from a healthy male epididymis cDNA library have been sequenced. The sequencing data provided 8,234 known sequences and 650 unknown cDNA fragments. Hundred and six of 650 unknown cDNA clone inserts were randomly selected for fully sequencing. There were 25 unknown unique sequences and 19 released but unreported sequences came out. By northern blot analysis, four sequences randomly selected from the 19 released sequences with no known function showed positive mRNA signals in epididymis and testis. The signals for three of six from those unknown group showed as epididymis abundant in a region-specific manner but not in the testis and other tissues tested. All the sequencing data will be available on the website www.sdscli.com. PMID- 18390569 TI - Microvascular function is impaired in ankylosing spondylitis and improves after tumour necrosis factor alpha blockade. AB - OBJECTIVES: Ankylosing spondylitis (AS) is associated with increased cardiovascular morbidity and mortality. Microvascular function has been linked to several risk factors for cardiovascular disease. Inflammation in AS may cause microvascular dysfunction. To test this, we assessed microvascular function in (a) patients with AS compared to healthy controls and (b) patients with AS before and after 1 month of anti-tumour necrosis factor (TNF)alpha treatment with etanercept. METHODS: A total of 15 consecutive patients with AS, who were scheduled for etanercept treatment according to the Assessment in Ankylosing Spondylitis (ASAS) group guidelines, and 12 healthy controls matched for age and sex, were recruited. Endothelium-dependent and independent vasodilatation in skin were evaluated with laser Doppler fluxmetry after iontophoresis of acetylcholine and sodium nitroprusside, respectively. Videomicroscopy was used to measure recruitment of skin capillaries after arterial occlusion. RESULTS: Compared to healthy controls, patients with AS had impaired endothelium-dependent vasodilatation and capillary recruitment. Following anti-TNFalpha treatment, microvascular function improved significantly for endothelium-dependent vasodilatation (p = 0.03) and capillary recruitment (p = 0.006). A significant correlation was observed between changes in endothelium-dependent vasodilatation and changes in erythrocyte sedimentation rate (ESR) (r = -0.56; p = 0.03). CONCLUSION: Microvascular dysfunction is present in patients with AS with active disease, but improves as inflammation regresses after TNFalpha blockade. PMID- 18390570 TI - Reliability of ultrasonography in detecting shoulder disease in patients with rheumatoid arthritis. AB - OBJECTIVE: To assess the intra and interobserver reproducibility of musculoskeletal ultrasonography (US) among rheumatologists in detecting destructive and inflammatory shoulder abnormalities in patients with rheumatoid arthritis (RA) and to determine the overall agreement between US and MRI. METHODS: A total of 14 observers examined 5 patients in 2 rounds independently and blindly of each other. US results were compared with MRI. Overall agreement of all findings, of positive findings on MRI, as well as intra and interobserver reliabilities, were calculated. RESULTS: Overall agreement between US and MRI was seen in 79% with regard to humeral head erosions (HHE), in 64% with regard to posterior recess synovitis (PRS), in 31% with regard to axillary recess synovitis (ARS), in 64% with regard to bursitis, in 50% with regard to biceps tenosynovitis (BT), and in 84% for complete cuff tear (CCT). Intraobserver and interobserver kappa was 0.69 and 0.43 for HHE, 0.29 and 0.49 for PRS, 0.57 and 1.00 for ARS, 0.17 and 0.51 for bursitis, 0.17 and 0.46 for BT and 0.52 and 0.6 for CCT, respectively. The intraobserver and interobserver kappa for power Doppler (PD) was 0.90 and 0.70 for glenohumeral signals and 0.60 and 0.51 for bursal signals, respectively. CONCLUSIONS: US is a reliable imaging technique for most shoulder pathology in RA especially with regard to PD. Standardisation of scanning technique and definitions of particular lesions may further enhance the reliability of US investigation of the shoulder. PMID- 18390571 TI - Crystals of monosodium urate monohydrate enhance lipopolysaccharide-induced release of interleukin 1 beta by mononuclear cells through a caspase 1-mediated process. AB - OBJECTIVE: Recent studies suggest that crystals of monosodium urate (MSU), deposited in joints of patients with acute gouty arthritis, activate the NACHT domain, leucine-rich repeat and pyrin domain-containing protein (NALP)3 inflammasome. In the present study we have investigated whether production of proinflammatory cytokines by crystals was exacerbated during costimulation with Toll-like receptor (TLR) ligands. METHODS: Mononuclear cells of 22 healthy donors were stimulated by various concentrations of MSU crystals in the absence or presence of lipopolysaccharide (LPS), Pam3Cys and flagellin. Production of tumour necrosis factor alpha (TNFalpha), interleukin (IL)1 beta and IL6, as well as the intracellular concentrations of proIL1 beta were measured by ELISA. mRNA transcripts of TNFalpha and IL1 beta were assessed by real-time PCR. Stimulation experiments were also performed with peripheral blood mononuclear cells (PBMCs) of one patient carrying a NALP3 mutation. RESULTS: MSU induced a moderate release of IL1 beta and IL6, but not of TNFalpha. Urate crystals amplified IL1 beta production stimulated by the TLR4 ligand LPS, while no synergy was apparent for IL6 production. In addition, no synergy between urate crystals and Pam3Cys (TLR2 ligand) or flagellin (TLR5 ligand) was apparent. The synergy between urate crystals and LPS was directed at the level of the NALP3 inflammasome, as it was present only when active IL1 beta was measured, but not at the level of IL1 mRNA or proIL1 beta. The synergy between LPS and MSU crystals ceased to exist in the presence of a caspase 1 inhibitor. CONCLUSIONS: MSU crystals act in synergy with LPS for the induction of enhanced release of IL1 beta. Increased cleavage of proIL1 beta by urate-activated caspase 1 is proposed as the underlying mechanism. PMID- 18390572 TI - Investigating a catalytic mechanism of hyperthermophilic L-threonine dehydrogenase from Pyrococcus horikoshii. AB - Based on our first structural data of L-threonine dehydrogenase (TDH) of Pyrococcus horikoshii (PhTDH), we examined its catalytic mechanism. The structural analysis indicated that a catalytic zinc atom at the active centre of PhTDH is coordinated by four residues (Cys42, His67, Glu68 and Glu152) with low affinity. These residues are highly conserved in alcohol dehydrogenases (ADHs) and TDHs. Several PhTDH mutants were prepared with respect to Glu152 and other residues, relating to the proton relay system that is substantially a rate limiting step in ADH. It was found that the E152D mutant showed 3-fold higher turnover rate and reduced affinities toward L-threonine and NAD(+), compared to wild-type PhTDH. The kinetic analysis of Glu152 mutants indicated that the carboxyl group of Glu152 is important for expressing the catalytic activity. The results obtained from pH dependency of kinetic parameters suggested that Glu152 to Asp substitution causes the enhancement of deprotonation of His47 or ionization of zinc-bound water and threonine in the enzyme-NAD(+) complex. Furthermore, it was predicted that the access of threonine substrate to the enzyme-NAD(+) complex induces a large conformational change in the active domain of PhTDH. From these results, we propose here that the proton relay system works as a catalytic mechanism of PhTDH. PMID- 18390573 TI - Concept, strategy and realization of lectin-based glycan profiling. AB - Lectins are a diverse group of carbohydrate-binding proteins. Each lectin has its own specificity profile. It is believed that lectins exist in all living organisms that produce glycans. From a practical viewpoint, lectins have been used extensively in biochemical fields including proteomics due to their usefulness as detection and enrichment tools for specific glycans. Nevertheless, they have often been underestimated as probes, especially compared with antibodies, because of their low affinity and broad specificity. However, together with the concept of glycomics, such properties of lectins are now considered to be suitable for the task of 'profiling' in order to cover a wider range of ligands. Recently there has been rapid movement in the field of proteomics aimed at the investigation of glycan-related biomarkers. This is partly because of limitations of the present approach of simply following changes in protein-level expression, without paying sufficient attention to the fact and effects of glycosylation. The trend is reflected in the frequent use of lectins in the contexts of glycoprotein enrichment and glycan profiling. However, there are many aspects to be considered in using lectins, which differ considerably from antibodies. In this article, the author, as a developer of two unique methodologies, frontal affinity chromatography (FAC) and the lectin microarray, describes critical points concerning the use of lectins, together with the concept, strategy and means to achieve advances in these emerging glycan profiling technologies. PMID- 18390574 TI - Carbon fibre composites: integrated electrochemical sensors for wound management. AB - The applicability of employing a carbon fibre mesh as an electrochemical sensing substructure for assessing urate transformations within wound exudates is evaluated. Prototype sensor assemblies have been designed and their response characteristics towards uric acid and other common physiological components are detailed. Modification of the carbon fibre sensor through surface anodization and the application of cellulose acetate permselective barriers have been shown to lead to optimized responses and much greater sensitivity (1440% increase) and specificity. These could enable the accurate periodic monitoring of uric acid in wound fluid. The performance characteristics of the composite sensors in whole blood, serum and blister fluid have been investigated. PMID- 18390575 TI - Development and validation of a simple NAFLD clinical scoring system for identifying patients without advanced disease. AB - BACKGROUND: Clinical predictors of advanced non-alcoholic liver disease (NAFLD) are needed to guide diagnostic evaluation and treatment. METHODS: To better understand the demographics of NAFLD and risk factors for advanced disease, this study analysed 827 patients with NAFLD at two geographically separate tertiary medical centres. RESULTS: The cohort was 51% female and had a median body mass index (BMI) of 33 kg/m(2); 3% had a normal BMI. Common co-morbidities included hypertension (60%) and diabetes (35%); insulin resistance was present in 91% and advanced fibrosis in 24% of patients. When comparing patients with no fibrosis or mild fibrosis to those with advanced fibrosis, BMI > or = 28 kg/m(2), age > 50 years, and aspartate transaminase/alanine aminotransferase (AST/ALT) ratio > or = 0.8, a quantitative assessment check index (QUICKI) score < 0.294 (equivalent to homeostasis model assessment (HOMA) > 6.2) and the presence of diabetes mellitus (DM) were individually associated by univariate analysis with odds ratios (ORs) of > or = 2.4 for advanced fibrosis. Based on the results of forced entry logistic regression analysis, three variables were combined in a weighted sum (BMI > or = 28 = 1 point, AAR of > or = 0.8 = 2 points, DM = 1 point) to form an easily calculated composite score for predicting advanced fibrosis called the BARD score. A score of 2-4 was associated with an OR for advanced fibrosis of 17 (confidence interval 9.2 to 31.9) and a negative predictive value of 96%. CONCLUSIONS: Insulin resistance and its co-morbidities are often present in patients with NAFLD. An easily calculated score based on readily available clinical data can reliably exclude the presence of advanced fibrosis in these patients, particularly among non-diabetics. PMID- 18390576 TI - Using support vector machine combined with auto covariance to predict protein protein interactions from protein sequences. AB - Compared to the available protein sequences of different organisms, the number of revealed protein-protein interactions (PPIs) is still very limited. So many computational methods have been developed to facilitate the identification of novel PPIs. However, the methods only using the information of protein sequences are more universal than those that depend on some additional information or predictions about the proteins. In this article, a sequence-based method is proposed by combining a new feature representation using auto covariance (AC) and support vector machine (SVM). AC accounts for the interactions between residues a certain distance apart in the sequence, so this method adequately takes the neighbouring effect into account. When performed on the PPI data of yeast Saccharomyces cerevisiae, the method achieved a very promising prediction result. An independent data set of 11,474 yeast PPIs was used to evaluate this prediction model and the prediction accuracy is 88.09%. The performance of this method is superior to those of the existing sequence-based methods, so it can be a useful supplementary tool for future proteomics studies. The prediction software and all data sets used in this article are freely available at http://www.scucic.cn/Predict_PPI/index.htm. PMID- 18390577 TI - Transcriptional regulation of chondrogenesis by coactivator Tip60 via chromatin association with Sox9 and Sox5. AB - Sox9 is a transcription factor of the SRY family required for several steps of chondrogenesis. It activates the expression of various chondrocyte-specific genes, but the mechanisms and role of cofactors involved in Sox9-regulated gene transcription are not fully understood. Here, we report on the characterization of a Tat interactive protein-60 (Tip60) as Sox9-associated protein identified in a yeast two-hybrid screen. Both in vitro and in vivo assays confirmed the specificity of interactions between Sox9 and Tip60 including the existence of an endogenous complex containing both polypeptides in chondrocytes. Gel shift assays showed the presence of a complex containing Sox9, Tip60 and the DNA of an enhancer region of the Col2a1 promoter. Reporter assays using a Col2a1 promoter with multimerized Col2a1 Sox9-binding sites indicated that Tip60 enhanced the transcriptional activity of Sox9. A larger Col2a1 promoter showed that Tip60 increased the activity of this promoter in the presence of both Sox9 and Sox5. Ectopic expression of Sox9 and transient-cotransfection with Tip60 in COS7 cells showed a more diffuse subnuclear colocalization, suggesting changes in the chromatin structure. Chromatin immunoprecipitation assays showed that Tip60, Sox9 and Sox5 associated with the same Col2a1 enhancer region. Consistent with a role of Tip60 in chondrogenesis, addition of Tip60 siRNA to limb-bud micromass cultures delayed chondrocyte differention. Tip60 enhances acetylation of Sox9 mainly through K61, 253, 398 residues; however, the K61/253/398A mutant of Sox9 still exhibited enhanced transcriptional activity by Tip60. Our results support the hypothesis that Tip60 is a coactivator of Sox9 in chondrocytes. PMID- 18390578 TI - Computational screen for spliceosomal RNA genes aids in defining the phylogenetic distribution of major and minor spliceosomal components. AB - The RNA molecules of the spliceosome are critical for specificity and catalysis during splicing of eukaryotic pre-mRNA. In order to examine the evolution and phylogenetic distribution of these RNAs, we analyzed 149 eukaryotic genomes representing a broad range of phylogenetic groups. RNAs were predicted using high sensitivity local alignment methods and profile HMMs in combination with covariance models. The results provide the most comprehensive view so far of the phylogenetic distribution of spliceosomal RNAs. RNAs were predicted in many phylogenetic groups where these RNA were not previously reported. Examples are RNAs of the major (U2-type) spliceosome in all fungal lineages, in lower metazoa and many protozoa. We also identified the minor (U12-type) spliceosomal U11 and U6atac RNAs in Acanthamoeba castellanii, where U12 spliceosomal RNA as well as minor introns were reported recently. In addition, minor-spliceosome-specific RNAs were identified in a number of phylogenetic groups where previously such RNAs were not observed, including the nematode Trichinella spiralis, the slime mold Physarum polycephalum and the fungal lineages Zygomycota and Chytridiomycota. The detailed map of the distribution of the U12-type RNA genes supports an early origin of the minor spliceosome and points to a number of occasions during evolution where it was lost. PMID- 18390579 TI - A national longitudinal study of the association between hours of TV viewing and the trajectory of BMI growth among US children. AB - OBJECTIVES: To assess the association between hours of TV viewing and the trajectory of BMI growth from Kindergarten to Grade 5 among a national longitudinal cohort of 7,334 US children. METHODS: Multilevel growth curve modeling was used to estimate children's BMI growth trajectories as a function of hours of TV viewing over time while controlling for gender, race/ethnicity, SES, birth weight, and baseline age. RESULTS: Hours of TV viewing were significantly positively associated with the acceleration of BMI growth from Kindergarten to Grade 5. CONCLUSIONS: Hours spent watching TV may be contributing to the recent dramatic increase in the prevalence of overweight and obesity among children. PMID- 18390580 TI - Analysis of iTRAQ data using Mascot and Peaks quantification algorithms. AB - The field of proteomics has been developing rapidly toward quantification of proteins. Despite the variety of experimental techniques available for peptide and protein labelling, there are few commercially available analytical tools with the ability to interpret data from any mass spectrometer. In this study, we compare two software packages, Mascot and Peaks, for the analysis of iTRAQ data from ESI-Q/TOF mass spectrometry. In the case of a six-protein mixture combined in a known proportion, the output of the Peaks algorithm deviated from the correct result by 14% on average, while the error of the Mascot quantification was nearly 200%. When the software were used to analyse iTRAQ data from a complex protein sample, the quantification results agreed within 20% for only 26% of the quantified proteins, showing significant differences in the two quantification algorithms. This comparison and analysis revealed major intricacies in peptide and protein quantification that must be taken into consideration for software development. PMID- 18390581 TI - Rituximab use in everyday clinical practice as a first-line biologic therapy for the treatment of DMARD-resistant rheumatoid arthritis. AB - OBJECTIVES: This study assessed the utility of rituximab for the therapy of RA in a non-academic environment in a group of cases where anti-TNF was either not available or relatively contraindicated. METHODS: Thirty-nine patients with active RA who had failed at least one DMARD received two rituximab infusions 2 weeks apart. Seventeen patients received two 1000 mg doses, and 22 received the 500 mg regimen. The 28-joint disease activity index (DAS28) and European League against Rheumatism (EULAR) response criteria were recorded at baseline, 3, 6, 9 and 12 months. RF and ANA were recorded at baseline and at 6 and 12 months. RESULTS: There was a significant improvement in the DAS28 at all time points, and EULAR response was observed in 29 of 33 patients (87.9%) at 3 months, 25 of 33 patients (75.8%) at 6 months, 22 of 29 patients (75.9%) at 9 months and 23 of 30 patients (76.7) at 12 months. Improvement was also noted in CRP, and both RF and ANA were generally reduced after treatment. Patients who were on the higher regimen of two 1000 mg doses appeared to respond slightly better compared with the lower dose regimen. CONCLUSIONS: Rituximab is well tolerated in everyday clinical practice and may represent a good short-term treatment option where anti TNF therapy is either unavailable or relatively contraindicated. PMID- 18390582 TI - Is FDG-PET useful in the evaluation of steroid-resistant PMR patients? PMID- 18390583 TI - Heberden's nodes and what Heberden could not see: the pivotal role of ligaments in the pathogenesis of early nodal osteoarthritis and beyond. AB - Despite its relatively high prevalence, polyarticular nature, limited treatment options and recognized genetic contribution, the study of generalized OA (GOA) has lagged behind that of isolated knee OA. Whilst the pathogenesis of OA has been viewed in relation to either articular cartilage or bone disease, this article offers a viewpoint on why GOA may, in fact, be primarily a disorder of ligaments, and to a lesser extent tendon and joint capsule dysfunction. A relatively fast presentation of GOA, typically in the perimenopausal period, and its recognition on clinical grounds alone makes this type of OA potentially useful for pathogenic studies in OA, in general. The recent high-resolution MRI studies, microanatomical studies and animal models, in addition to established clinical and radiographic data that support this ligament-centric perspective of disease, are reviewed. The earliest structural abnormalities in GOA may be evident in ligaments and the ligament-associated 'enthesis organ', where degenerative changes are evident. Ligaments also influence the expression of joint damage including Heberden's node and joint erosion formation. Joint inflammation in a 'periarthritis' pattern is well recognized in GOA, and histological studies have shown that the ligament and capsule could represent the epicentre of such inflammatory changes. A perspective is also offered on how ligaments could play a pivotal role in OA in general; for example, the loss of joint space in knee OA due to meniscal extrusion could ultimately be related to derangement of the medial collateral ligament to which the meniscus is anchored. PMID- 18390585 TI - Evaluation of a new algorithm in classification of systemic vasculitis. AB - OBJECTIVE: Recently, a new consensus algorithm for classification of ANCA associated vasculitis (AAV) and PAN had been proposed by Watt et al. for epidemiological studies. In order to evaluate this algorithm, the current study used the algorithm to reclassify the patients with AAV and PAN in our centre. METHODS: Five hundred and fifty Chinese patients with primary systemic vasculitis diagnosed in our referral diagnostic centre during the past 10 years were retrospectively studied. Four hundred and ninety-three out of 550 were ANCA positive. We compared the new consensus algorithm and the 1994 Chapel Hill Consensus Conference (CHCC) definitions supplemented with surrogate parameters, in the same cohort of patients with primary systemic vasculitis. RESULTS: Applying the CHCC definitions with surrogate parameters, the diagnoses were Churg Strauss syndrome (CSS) (n = 0), WG (n = 127), microscopic polyangiitis (MPA) (n = 363), PAN (n = 4) and unclassified (n = 56). Using the new consensus algorithm, the diagnoses were CSS (n = 2), WG (n = 199), MPA (n = 329), PAN (n = 0) and unclassified (n = 20). CONCLUSIONS: Watts' algorithm was a useful method to classify patients into a single category, with less unclassified patients and without overlapping diagnosis, which allows their use in epidemiological studies. PMID- 18390584 TI - Increased IL-4+ CD8+ T cells in peripheral blood and autoreactive CD8+ T cell lines of patients with inflammatory arthritis. AB - OBJECTIVE: To measure the frequencies of IL-4+ CD8+ T cells from patients with AS and RA, and to assess their clinical relevance and properties. METHODS: Peripheral blood (PB) and clinical data were obtained from 37 AS, 36 RA patients and 37 healthy controls. We also generated IL-4-producing CD8+ T cell lines and clones by co-culture with autologous dendritic cells. Using flow cytometry, we evaluated intracellular cytokine expression by T cells following stimulation with PMA and calcium ionophore. The phenotype and ability of the IL-4-producing CD8+ T cell clones to suppress IFN-gamma production were examined. RESULTS: The percentages of IL-4+ CD8+ T cells were higher in PB of patients with AS and RA than controls (medians 0.90 and 0.84% vs 0.30%). In RA, patients with active inflammation had an increased percentage of IL-4+ CD8+ T cells. Higher frequencies of IL-4+ CD8+ T cells were also found in CD8+ T cell lines established from patients with arthritis. Interestingly, most IL-4+ CD8+ T cells produced TNF-alpha. Cloning the CD8+ T cell lines yielded more IL-4-producing clones from AS (23%) and RA patients (14%) than from controls (7%). The ability to suppress IFN-gamma production was observed in 56% (AS) and 85% (RA) of IL-4 producing clones. Suppressive IL-4+ CD8+ T cell clones from RA patients showed a similar regulatory phenotype to the clones previously isolated from AS patients. CONCLUSIONS: Expansion of IL-4+ CD8+ T cells, which may include precursors of a regulatory CD8+ T cell subset, may represent a general response to chronic joint inflammation. PMID- 18390586 TI - Abnormal digital neurovascular response to local heating in systemic sclerosis. AB - OBJECTIVES: To investigate neurovascular dysfunction using the axon reflex dependent hyperaemia (initial peak of skin local heating response) in fingers of patients with SSc or primary RP. METHODS: Ten healthy subjects were initially enrolled to compare axon reflex-dependent thermal hyperaemia between the finger and forearm cutaneous circulations. Then, 10 patients with primary RP and 16 patients with SSc participated in a similar protocol focusing on the finger circulation only. Lidocaine/prilocaine cream was applied for 1 h to produce local blockade of cutaneous sensory nerves. After lidocaine/prilocaine pre-treatment, laser Doppler probes were heated from skin temperature to 42 degrees C for 30 min, and 44 degrees C for 5 min to achieve maximal skin blood flow. Data were expressed as a percentage of maximal cutaneous vascular conductance. RESULTS: In healthy volunteers, we observed a significantly higher initial peak on the finger compared with the forearm, with both responses blunted following topical anaesthesia. In primary RP patients, we observed a decreased initial peak following lidocaine/prilocaine pre-treatment in the finger circulation [96.7% (33.4) vs 75.9% (29.5) with anaesthesia, P = 0.02]. In contrast, pre-treatment did not alter the initial peak in patients with SSc. A minute-by-minute analysis showed no delay of the initial peak. CONCLUSIONS: We show an abnormal digital neurovascular response to local heating in SSc. Thermal hyperaemia could be monitored as a clinical test for neurovascular function in SSc. Further studies are required to test whether the abnormal digital neurovascular response correlates to the degree of peripheral vascular involvement. PMID- 18390587 TI - Glucocorticoid treatment inhibits annexin-1 expression in rheumatoid arthritis CD4+ T cells. AB - OBJECTIVE: Annexin-1 (Anx-A1) has been recently shown to play a key role in T cell activation and to be highly expressed in T cells from RA patients. Here, we investigated the effects of glucocorticoids (GCs) on Anx-A1 expression in T cells in vitro and in vivo. METHODS: To evaluate the effects of dexamethasone (Dex) on Anx-A1 expression, human peripheral blood T cells were incubated with Dex and then analysed by real-time PCR and western blotting. Similar experiments were carried out in vivo by measuring Anx-A1 levels in T cells from patients with RA before and after administration of steroids. RESULTS: Incubation of T cells with Dex decreased Anx-A1 levels in a time-dependent fashion and almost abolished its expression after 12 h. Stimulation of T cells pre-incubated with Dex for 12 h with anti-CD3/CD28 led to significant reduction of IL-2 production. Addition of human recombinant Anx-A1 to Dex-treated cells reversed the inhibitory effects of the steroids on anti-CD3/CD28-induced IL-2 production. Treatment of RA patients with steroid decreased Anx-A1 expression in T cells. CONCLUSIONS: GCs suppress Anx-A1 expression in T cells in vitro and in vivo. These results provide evidence for a novel pathway by which steroids regulate the adaptive immune response and suggest that Anx-A1 may represent a target for the treatment of autoimmune diseases. PMID- 18390588 TI - Increased risk of myeloid leukaemia in patients with ankylosing spondylitis following treatment with radium-224. AB - OBJECTIVE: To investigate long-term health effects in AS patients treated with (224)Ra. METHODS: A prospective epidemiological study has been carried out on 1471 AS patients treated with repeated intravenous injections of (224)Ra between 1948 and 1975. These patients have been followed together with a control group of 1324 AS patients not treated with radioactive drugs and/or X-rays. Numbers of malignancies expected in a normal population were computed from German and Danish cancer registry data. RESULTS: After a mean follow-up time of 26 yrs in the exposed group or 25 yrs in the control group, causes of death have been ascertained for 1006 exposed patients and 1072 controls. In particular, 19 cases of leukaemia were observed in the exposure group (vs 6.8 cases expected, P < 0.001) compared to 12 cases of leukaemia in the control group (vs 7.5 cases expected). Further subclassification of the leukaemia cases demonstrated a high increase of myeloid leukaemia in the exposure group (11 cases observed vs 2.9 cases expected, P < 0.001), especially a high excess of acute myeloid leukaemias (7 cases observed vs 1.8 cases expected, P = 0.003), whereas in the controls the observed cases are within the expected range (4 myeloid leukaemias vs 3.1 cases expected). CONCLUSIONS: The enhanced leukaemia incidence in the exposed group is in line with results from experiments in mice injected with varying amounts of the bone-seeking alpha-emitter (224)Ra. In these studies, in animals exposed to lower doses of (224)Ra, i.e. at doses lower than those found to induce osteosarcomas, an increased risk of leukaemia was observed. PMID- 18390590 TI - Revised classification of lupus nephritis is valuable in predicting renal outcome with an indication of the proportion of glomeruli affected by chronic lesions. AB - OBJECTIVES: To determine if the International Society of Nephrology/Renal Pathology Society (ISN/RPS) 2003 classification of lupus nephritis (LN) is helpful in predicting renal outcome. METHODS: A total of 92 patients with LN who underwent renal biopsy in our hospital were re-classified according to the ISN/RPS 2003 criteria. RESULTS: The mean patient age was 36.8 yrs and the median observation period was 65 months. The relative frequency for each class was as follows: Class I (minimal mesangial LN) 0%, Class II (mesangial proliferative LN) 13%, Class III (focal LN) 17%, Class IV (diffuse LN) 60% and Class V (membranous LN) 10%. Within Class IV, diffuse segmental (Class IV-S) was 25% and diffuse global (Class IV-G) 75%. During the observation period, renal function was more likely to deteriorate in Class IV-G cases than in Class IV-S cases. Importantly, when Class IV-G was subdivided into cases involving active lesion alone [IV-G (A)] or chronic lesion [IV-G (A/C)], the majority of cases in IV-G (A) was nephrotic, but responded well to therapy. In contrast, renal function declined only in IV-G (A/C) cases. Patients with Class IV-G (A/C) had persistent proteinuria in spite of intensified therapies. Moreover, the higher proportion of chronic lesions was related with the deterioration of renal function. CONCLUSIONS: This study showed that in Class IV-G cases, renal outcome differed in the presence of chronicity. Chronicity could be a critical factor in predicting outcome. Thus, the revised classification of LN is clinically valuable in identifying different renal outcomes among patients with diffuse LN. PMID- 18390589 TI - Rheumatoid arthritis patients who smoke have a higher need for DMARDs and feel worse, but they do not have more joint damage than non-smokers of the same serological group. AB - OBJECTIVES: To investigate the influence of smoking on disease activity, drug need and radiographic joint damage in RF-positive and -negative patients with early RA. METHODS: Baseline and 3-yr follow-up data of 896 patients of an early RA cohort comprised clinical and radiographic parameters (Ratingen Score). Information about disease severity, treatment and smoking were obtained by questionnaires. Univariate and multivariate analyses were used to show the influence of smoking on drug use, ACR improvement and joint damage. Smokers and non-smokers were compared according to RF serology. RESULTS: Fifty per cent of the patients were never, 23% past and 27% current smokers. Current smokers were significantly more often RF-positive (71%) than past (66%) or never smokers (53%), but neither the RF-positive nor the RF-negative current smokers had higher 28-joint disease activity score (DAS28) or radiographic scores than never or past smokers. Within 3 yrs, current smokers had taken significantly more DMARD combinations or biologics. Non-smokers and those with <20 pack-years (PYs) had a 2-fold higher probability to reach ACR improvement than heavy smokers (>20 PYs). However, smokers did not differ in radiographic joint damage when compared with non-smokers of the same serological group. CONCLUSIONS: The higher use of DMARDs may indicate that smoking weakens the potency of anti-rheumatic drugs and/or is needed to control an otherwise higher disease activity. Since the risk of adverse events increases with the amount of drugs taken, this is another reason to persuade RA patients to quit smoking. PMID- 18390591 TI - Interaction of KNAT6 and KNAT2 with BREVIPEDICELLUS and PENNYWISE in Arabidopsis inflorescences. AB - The three amino acid loop extension (TALE) homeodomain superfamily, which comprises the KNOTTED-like and BEL1-like families, plays a critical role in regulating meristem activity. We previously demonstrated a function for KNAT6 (for KNOTTED-like from Arabidopsis thaliana 6) in shoot apical meristem and boundary maintenance during embryogenesis. KNAT2, the gene most closely related to KNAT6, does not play such a role. To investigate the contribution of KNAT6 and KNAT2 to inflorescence development, we examined their interactions with two TALE genes that regulate internode patterning, BREVIPEDICELLUS (BP) and PENNYWISE (PNY). Our data revealed distinct and overlapping interactions of KNAT6 and KNAT2 during inflorescence development. Removal of KNAT6 activity suppressed the pny phenotype and partially rescued the bp phenotype. Removal of KNAT2 activity had an effect only in the absence of both BP and KNAT6 or in the absence of both BP and PNY. Consistent with this, KNAT6 and KNAT2 expression patterns were enlarged in both bp and pny mutants. Thus, the defects seen in pny and bp are attributable mainly to the misexpression of KNAT6 and to a lesser extent of KNAT2. Hence, our data showed that BP and PNY restrict KNAT6 and KNAT2 expression to promote correct inflorescence development. This interaction was also revealed in the carpel. PMID- 18390592 TI - Phosducin-Like Protein 3 is required for microtubule-dependent steps of cell division but not for meristem growth in Arabidopsis. AB - Given the central role of cell division in meristems, one might expect meristem growth to be regulated by mitotic checkpoints, including checkpoints for correct microtubule function. Here, we studied the role of two close Phosducin-Like Protein 3 homologs from Arabidopsis thaliana (PLP3a and PLP3b) in the microtubule assembly pathway and determined the consequences of inhibiting PLP3a and PLP3b expression in the meristem. PLP3 function is essential in Arabidopsis: impairing PLP3a and PLP3b expression disrupted microtubule arrays and caused polyploidy, aneuploidy, defective cytokinesis, and disoriented cell growth. Consistent with a role in microtubule formation, PLP3a interacted with beta-tubulin in the yeast two-hybrid assay and, when overexpressed, increased resistance to drugs that inhibit tubulin polymerization. Inhibition of PLP3 function targeted to the meristem caused severe mitotic defects, but the cells carried on cycling through DNA replication and abortive cytokinesis. Thus, we showed that PLP3 is involved in microtubule formation in Arabidopsis and provided genetic evidence that cell viability and growth in the meristem are not subordinate to successful completion of microtubule-dependent steps of cell division. PMID- 18390593 TI - Conserved C-terminal motifs required for avirulence and suppression of cell death by Phytophthora sojae effector Avr1b. AB - The sequenced genomes of oomycete plant pathogens contain large superfamilies of effector proteins containing the protein translocation motif RXLR-dEER. However, the contributions of these effectors to pathogenicity remain poorly understood. Here, we show that the Phytophthora sojae effector protein Avr1b can contribute positively to virulence and can suppress programmed cell death (PCD) triggered by the mouse BAX protein in yeast, soybean (Glycine max), and Nicotiana benthamiana cells. We identify three conserved motifs (K, W, and Y) in the C terminus of the Avr1b protein and show that mutations in the conserved residues of the W and Y motifs reduce or abolish the ability of Avr1b to suppress PCD and also abolish the avirulence interaction of Avr1b with the Rps1b resistance gene in soybean. W and Y motifs are present in at least half of the identified oomycete RXLR-dEER effector candidates, and we show that three of these candidates also suppress PCD in soybean. Together, these results indicate that the W and Y motifs are critical for the interaction of Avr1b with host plant target proteins and support the hypothesis that these motifs are critical for the functions of the very large number of predicted oomycete effectors that contain them. PMID- 18390596 TI - Synthetic nanopores as a test case for ion channel theories: the anomalous mole fraction effect without single filing. AB - The predictions of a theory for the anomalous mole fraction effect (AMFE) are tested experimentally with synthetic nanopores in plastic. The negatively charged synthetic nanopores under consideration are highly cation selective and 50 A in diameter at their smallest point. These pores exhibit an AMFE in mixtures of Ca(2+) and monovalent cations. An AMFE occurs when the conductance through a pore is lower in a mixture of salts than in the pure salts at the same concentration. For ion channels, the textbook interpretation of the AMFE is that multiple ions move through the pore in coordinated, single-file motion. However, because the synthetic nanopores are so wide, their AMFE shows that single filing is not necessary for the AMFE. It is shown that the AMFE in the synthetic nanopores is explained by a theory of preferential ion selectivity. The unique properties of the synthetic nanopores allow us to experimentally confirm several predictions of this theory. These same properties make synthetic nanopores an interesting new platform to test theories of ion channel permeation and selectivity in general. PMID- 18390594 TI - Beta-AMYLASE4, a noncatalytic protein required for starch breakdown, acts upstream of three active beta-amylases in Arabidopsis chloroplasts. AB - This work investigated the roles of beta-amylases in the breakdown of leaf starch. Of the nine beta-amylase (BAM)-like proteins encoded in the Arabidopsis thaliana genome, at least four (BAM1, -2, -3, and -4) are chloroplastic. When expressed as recombinant proteins in Escherichia coli, BAM1, BAM2, and BAM3 had measurable beta-amylase activity but BAM4 did not. BAM4 has multiple amino acid substitutions relative to characterized beta-amylases, including one of the two catalytic residues. Modeling predicts major differences between the glucan binding site of BAM4 and those of active beta-amylases. Thus, BAM4 probably lost its catalytic capacity during evolution. Total beta-amylase activity was reduced in leaves of bam1 and bam3 mutants but not in bam2 and bam4 mutants. The bam3 mutant had elevated starch levels and lower nighttime maltose levels than the wild type, whereas bam1 did not. However, the bam1 bam3 double mutant had a more severe phenotype than bam3, suggesting functional overlap between the two proteins. Surprisingly, bam4 mutants had elevated starch levels. Introduction of the bam4 mutation into the bam3 and bam1 bam3 backgrounds further elevated the starch levels in both cases. These data suggest that BAM4 facilitates or regulates starch breakdown and operates independently of BAM1 and BAM3. Together, our findings are consistent with the proposal that beta-amylase is a major enzyme of starch breakdown in leaves, but they reveal unexpected complexity in terms of the specialization of protein function. PMID- 18390597 TI - Single-molecule force spectroscopy reveals a stepwise unfolding of Caenorhabditis elegans giant protein kinase domains. AB - Myofibril assembly and disassembly are complex processes that regulate overall muscle mass. Titin kinase has been implicated as an initiating catalyst in signaling pathways that ultimately result in myofibril growth. In titin, the kinase domain is in an ideal position to sense mechanical strain that occurs during muscle activity. The enzyme is negatively regulated by intramolecular interactions occurring between the kinase catalytic core and autoinhibitory/regulatory region. Molecular dynamics simulations suggest that human titin kinase acts as a force sensor. However, the precise mechanism(s) resulting in the conformational changes that relieve the kinase of this autoinhibition are unknown. Here we measured the mechanical properties of the kinase domain and flanking Ig/Fn domains of the Caenorhabditis elegans titin-like proteins twitchin and TTN-1 using single-molecule atomic force microscopy. Our results show that these kinase domains have significant mechanical resistance, unfolding at forces similar to those for Ig/Fn beta-sandwich domains (30-150 pN). Further, our atomic force microscopy data is consistent with molecular dynamic simulations, which show that these kinases unfold in a stepwise fashion, first an unwinding of the autoinhibitory region, followed by a two-step unfolding of the catalytic core. These data support the hypothesis that titin kinase may function as an effective force sensor. PMID- 18390598 TI - Equinatoxin II permeabilizing activity depends on the presence of sphingomyelin and lipid phase coexistence. AB - Equinatoxin II is a pore-forming protein of the actinoporin family. After membrane binding, it inserts its N-terminal alpha-helix and forms a protein/lipid pore. Equinatoxin II activity depends on the presence of sphingomyelin in the target membrane; however, the role of this specificity is unknown. On the other hand, sphingomyelin is considered an essential ingredient of lipid rafts and promotes liquid-ordered/liquid-disordered phase separation in model membranes that mimic raft composition. Here, we used giant unilamellar vesicles to simultaneously investigate the effect of sphingomyelin and phase separation on the membrane binding and permeabilizing activity of Equinatoxin II. Our results show that Equinatoxin II binds preferentially to the liquid-ordered phase over the liquid-disordered one and that it tends to concentrate at domain interfaces. In addition, sphingomyelin strongly enhances membrane binding of the toxin but is not sufficient for membrane permeabilization. Under the same experimental conditions, Equinatoxin II formed pores in giant unilamellar vesicles containing sphingomyelin only when liquid-ordered and -disordered phases coexisted. Our observations demonstrate the importance of phase boundaries for Equinatoxin II activity and suggest a double role of sphingomyelin as a specific receptor for the toxin and as a promoter of the membrane organization necessary for Equinatoxin II action. PMID- 18390599 TI - Dynamics and Cleavability at the alpha-cleavage site of APP(684-726) in different lipid environments. AB - The occurrence of late-onset Alzheimer's disease has been related to the lipid homeostasis. We tested whether the membrane lipid environment affects the dynamics and cleavability of a model peptide corresponding to the amino acid sequence 684-726 of the amyloid precursor protein APP reconstituted in liposomes. Solid-state NMR with (2)H-Ala(713), which is located within the putative transmembrane domain, suggested that the peptide observes less rotational motion in egg phosphatidylcholine (PhC) membranes than in dimyristoyl phosphatidylcholine (DMPC) bilayers above the main phase transition temperature T(c). The residue (15)N-Ala(692), which is in the vicinity of the alpha-cleavage site, i.e., Lys(687), showed less motion after reconstitution in distearoyl phosphatidylcholine liposomes ZZZ), which involves only small protein structural changes. PMID- 18390619 TI - Influence of lipid saturation grade and headgroup charge: a refined lung surfactant adsorption model. AB - Rapid adsorption of surfactant material to the air/liquid interface of the lung is essential for maintaining normal lung function. The detailed mechanism of this process, however, remains unclear. In this study, we elucidate the influence of lipid saturation grade and headgroup charge of surface layer lipids on surfactant protein (SP)-induced vesicle insertion into monolayers spread at the air/water interface of a film balance. We used dipalmitoylphosphatidlycholine (DPPC),1,2 dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG), 1-palmitoyl-2-oleoyl-sn-glycero 3-phosphocholine (POPC), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG) as monolayer lipids doped with either hydrophobic surfactant-specific protein SP-B or SP-C (0.2 and 0.4 mol %, respectively). Vesicles consisting of DPPC/DPPG (4:1, mol ratio) were injected into a stirred subphase to quantify adsorption kinetics. Based on kinetic film balance and fluorescence measurements, a refined model describing distinct steps of vesicle adsorption to surfactant monolayers is presented. First, in a protein-independent step, lipids from vesicles bridged to the interfacial film by Ca(2+) ions are inserted into defects of a disordered monolayer at low surface pressures. Second, in a SP-facilitated step, active material insertion involving an SP-B- or SP-C-induced flip-flop of lipids occurs at higher surface pressures. Negatively charged lipids obviously influence the threshold pressures at which this second protein-mediated adsorption mechanism takes place. PMID- 18390620 TI - More gating charges are needed to open a Shaker K+ channel than are needed to open an rBIIA Na+ channel. AB - This study presents what is, to our knowledge, a novel technique by means of which the ratio of the single gating charges of voltage-gated rat brain IIA (rBIIA) sodium and Shaker potassium ion channels was estimated. In the experiment, multiple tandems of enhanced green fluorescent protein were constructed and inserted into the C-terminals of Na(+) and K(+) ion channels. cRNA of Na(+) and K(+) ion channels was injected and expressed in Xenopus laevis oocytes. The two electrode voltage-clamp technique allowed us to determine the total gating charge of sodium and potassium ion channels, while a relative measure of the amount of expressed channels could be established on the basis of the quantification of the fluorescence intensity of membrane-bound channels marked by enhanced green fluorescent proteins. As a result, gating charge and fluorescence intensity were found to be positively correlated. A relative comparison of the single gating charges of voltage-gated sodium and potassium ion channels could thus be established: the ratio of the single gating charges of the Shaker potassium channel and the rBIIA sodium channel was found to be 2.5 +/- 0.4. Assuming the single channel gating charge of the Shaker K(+) channel to be approximately 13 elementary charges (well supported by other studies), this leads to approximately six elementary charges for the rBIIA sodium channel, which includes a fraction of gating charge that is missed during inactivation. PMID- 18390621 TI - Unique backbone-water interaction detected in sphingomyelin bilayers with 1H/31P and 1H/13C HETCOR MAS NMR spectroscopy. AB - Two-dimensional (1)H/(31)P dipolar heteronuclear correlation (HETCOR) magic-angle spinning nuclear magnetic resonance (NMR) is used to investigate the correlation of the lipid headgroup with various intra- and intermolecular proton environments. Cross-polarization NMR techniques involving (31)P have not been previously pursued to a great extent in lipid bilayers due to the long (1)H-(31)P distances and high degree of headgroup mobility that averages the dipolar coupling in the liquid crystalline phase. The results presented herein show that this approach is very promising and yields information not readily available with other experimental methods. Of particular interest is the detection of a unique lipid backbone-water intermolecular interaction in egg sphingomyelin (SM) that is not observed in lipids with glycerol backbones like phosphatidylcholines. This backbone-water interaction in SM is probed when a mixing period allowing magnetization exchange between different (1)H environments via the nuclear Overhauser effect (NOE) is included in the NMR pulse sequence. The molecular information provided by these (1)H/(31)P dipolar HETCOR experiments with NOE mixing differ from those previously obtained by conventional NOE spectroscopy and heteronuclear NOE spectroscopy NMR experiments. In addition, two-dimensional (1)H/(13)C INEPT HETCOR experiments with NOE mixing support the (1)H/(31)P dipolar HETCOR results and confirm the presence of a H(2)O environment that has nonvanishing dipolar interactions with the SM backbone. PMID- 18390622 TI - Ca2+-mobility in the sarcoplasmic reticulum of ventricular myocytes is low. AB - The sarcoplasmic reticulum (SR) in ventricular myocytes contains releasable Ca(2+) for activating cellular contraction. Recent measurements of intra-SR (luminal) Ca(2+) suggest a high diffusive Ca(2+)-mobility constant (D(CaSR)). This could help spatially to unify SR Ca(2+)-content ([Ca(2+)](SRT)) and standardize Ca(2+)-release throughout the cell. But measurements of localized depletions of luminal Ca(2+) (Ca(2+)-blinks), associated with local Ca(2+) release (Ca(2+)-sparks), suggest D(CaSR) may actually be low. Here we describe a novel method for measuring D(CaSR). Using a cytoplasmic Ca(2+)-fluorophore, we estimate regional [Ca(2+)](SRT) from localized, caffeine-induced SR Ca(2+) release. Caffeine microperfusion of one end of a guinea pig or rat myocyte diffusively empties the whole SR at a rate indicating D(CaSR) is 8-9 microm(2)/s, up to tenfold lower than previous estimates. Ignoring background SR Ca(2+) leakage in our measurement protocol produces an artifactually high D(CaSR) (>40 microm(2)/s), which may also explain the previous high values. Diffusion-reaction modeling suggests that a low D(CaSR) would be sufficient to support local SR Ca(2+)-signaling within sarcomeres during excitation-contraction coupling. Low D(CaSR) also implies that [Ca(2+)](SRT) may readily become spatially nonuniform, particularly under pathological conditions of spatially nonuniform Ca(2+) release. Local control of luminal Ca(2+), imposed by low D(CaSR), may complement the well-established local control of SR Ca(2+)-release by Ca(2+) channel/ryanodine receptor couplons. PMID- 18390623 TI - Liquid-liquid domains in bilayers detected by wide angle X-ray scattering. AB - Wide angle x-ray scattering (WAXS) from oriented lipid multilayers is used to examine liquid-ordered (Lo)/liquid-disordered (Ld) phase coexistence in the system 1,2-dioleoyl-sn-glycero-3-phosphocholine/1,2-dipalmitoyl-sn-glycero-3 phosphocholine/cholesterol (DOPC/DPPC/Chol), which is a model for the outer leaflet of the animal cell plasma membrane. Using the method of analysis developed in the accompanying work, we find that two orientational distributions are necessary to fit the WAXS data at lower temperatures, whereas only one distribution is needed at temperatures higher than the miscibility transition temperature, T(mix) = 25-35 degrees C (for 1:1 DOPC/DPPC with 15%, 20%, 25%, and 30% Chol). We propose that the necessity for two distributions is a criterion for coexistence of Lo domains with a high S(x-ray) order parameter and Ld domains with a lower order parameter. This criterion is capable of detecting coexistence of small domains or rafts that the conventional x-ray criterion of two lamellar D spacings may not. Our T(mix) values tend to be slightly larger than published NMR results and microscopy results when the fluorescence probe artifact is considered. This is consistent with the sensitivity of WAXS to very short time and length scales, which makes it more capable of detecting small, short-lived domains that are likely close to T(mix). PMID- 18390624 TI - Order parameters and areas in fluid-phase oriented lipid membranes using wide angle X-ray scattering. AB - We used wide angle x-ray scattering (WAXS) from stacks of oriented lipid bilayers to measure chain orientational order parameters and lipid areas in model membranes consisting of mixtures of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC)/cholesterol and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)/cholesterol in fluid phases. The addition of 40% cholesterol to either DOPC or DPPC changes the WAXS pattern due to an increase in acyl chain orientational order, which is one of the main properties distinguishing the cholesterol-rich liquid-ordered (Lo) phase from the liquid-disordered (Ld) phase. In contrast, powder x-ray data from multilamellar vesicles does not yield information about orientational order, and the scattering from the Lo and Ld phases looks similar. An analytical model to describe the relationship between the chain orientational distribution and WAXS data was used to obtain an average orientational order parameter, S(x-ray). When 40% cholesterol is added to either DOPC or DPPC, S(x-ray) more than doubles, consistent with previous NMR order parameter measurements. By combining information about the average chain orientation with the chain-chain correlation spacing, we extended a commonly used method for calculating areas for gel-phase lipids to fluid-phase lipids and obtained agreement to within 5% of literature values. PMID- 18390625 TI - Gating mechanisms of mechanosensitive channels of large conductance, II: systematic study of conformational transitions. AB - Part II of this study is based on the continuum mechanics-based molecular dynamics-decorated finite element method (MDeFEM) framework established in Part I. In Part II, the gating pathways of Escherichia coli-MscL channels under various basic deformation modes are simulated. Upon equibiaxial tension (which is verified to be the most effective mode for gating), the MDeFEM results agree well with both experiments and all-atom simulations in literature, as well as the analytical continuum models and elastic network models developed in Part I. Different levels of model sophistication and effects of structural motifs are explored in detail, where the importance of mechanical roles of transmembrane helices, cytoplasmic helices, and loops are discussed. The conformation transitions under complex membrane deformations are predicted, including bending, torsion, cooperativity, patch clamp, and indentation. Compared to atom-based molecular dynamics simulations and elastic network models, the MDeFEM framework is unusually well-suited for simulating complex deformations at large length scales. The versatile hierarchical framework can be further applied to simulate the gating transition of other mechanosensitive channels and other biological processes where mechanical perturbation is important. PMID- 18390626 TI - Gating mechanisms of mechanosensitive channels of large conductance, I: a continuum mechanics-based hierarchical framework. AB - A hierarchical simulation framework that integrates information from molecular dynamics (MD) simulations into a continuum model is established to study the mechanical response of mechanosensitive channel of large-conductance (MscL) using the finite element method (FEM). The proposed MD-decorated FEM (MDeFEM) approach is used to explore the detailed gating mechanisms of the MscL in Escherichia coli embedded in a palmitoyloleoylphosphatidylethanolamine lipid bilayer. In Part I of this study, the framework of MDeFEM is established. The transmembrane and cytoplasmic helices are taken to be elastic rods, the loops are modeled as springs, and the lipid bilayer is approximated by a three-layer sheet. The mechanical properties of the continuum components, as well as their interactions, are derived from molecular simulations based on atomic force fields. In addition, analytical closed-form continuum model and elastic network model are established to complement the MDeFEM approach and to capture the most essential features of gating. In Part II of this study, the detailed gating mechanisms of E. coli-MscL under various types of loading are presented and compared with experiments, structural model, and all-atom simulations, as well as the analytical models established in Part I. It is envisioned that such a hierarchical multiscale framework will find great value in the study of a variety of biological processes involving complex mechanical deformations such as muscle contraction and mechanotransduction. PMID- 18390627 TI - Measuring diffusion and binding kinetics by contact area FRAP. AB - The immunological synapse is a stable intercellular structure that specializes in substance and signal transfer from one immune cell to another. Its formation is regulated in part by the diffusion of adhesion and signaling molecules into, and their binding of countermolecules in the contact area. The stability of immunological synapses allows receptor-ligand interactions to approximate chemical equilibrium despite other dynamic aspects. We have developed a mathematical model that describes the coupled reaction-diffusion process in an established immunological synapse. In this study, we extend a previously described contact area fluorescence recovery after photobleaching (FRAP) experiment to test the validity of the model. The receptor binding activity and lateral mobility of fluorescently labeled, lipid-anchored ligands in the bilayer resulted in their accumulation, as revealed by a much higher fluorescence intensity inside the contact area than outside. After complete photobleaching of the synapse, fluorescence recovery requires ligands to dissociate and rebind, and to diffuse in and out of the contact area. Such a FRAP time course consequently provides information on reaction and diffusion, which can be extracted by fitting the model solution to the data. Surprisingly, reverse rates in the two dimensional contact area were at least 100-fold slower than in three-dimensional solution. As previously reported in immunological synapses, a significant nonrecoverable fraction of fluorescence was observed with one of two systems studied, suggesting some ligands either dissociated or diffused much more slowly compared with other ligands in the same synapse. The combined theory and experiment thus provides a new method for in situ measurements of kinetic rates, diffusion coefficients, and nonrecoverable fractions of interacting molecules in immunological synapses and other stable cell-bilayer junctions. PMID- 18390628 TI - A coupled diffusion-kinetics model for analysis of contact-area FRAP experiment. AB - Kinetic rates and binding affinity of receptor-ligand interactions are important determinants of cell adhesion. Measurements of these parameters in fluid phase using soluble molecules (i.e., three-dimensionial parameters) do not necessarily correlate with their counterparts measured when both binding partners are respectively anchored to two apposing surfaces (i.e., two-dimensional (2D) parameters). Moreover, 2D affinities measured by different methods can differ by orders of magnitude. Here we describe a coupled diffusion-reaction model for the fluorescence recovery after photobleaching experiment previously used to demonstrate the dynamics of adhesive bonds in the contact area. Applying the mathematical model to the contact area fluorescence recovery after photobleaching experiment enables in situ measurements of 2D kinetic rates of the adhesion molecules and their retarded diffusion in a stable contact area. The mathematical properties of the model are characterized in this article and its experimental validation will be presented in the companion article. PMID- 18390629 TI - Association between asthma and serum adiponectin concentration in women. AB - BACKGROUND: The association of murine asthma with adiposity may be mediated by adiponectin, an anti-inflammatory adipokine with reduced serum concentrations in obese subjects. A study was undertaken to examine whether the serum adiponectin concentration is associated with human asthma and whether it explains the association between adiposity and asthma, particularly in women and in premenopausal women. METHODS: A cross-sectional analysis was performed of 2890 eligible subjects at year 15 of the Coronary Artery Risk Development in Young Adults (CARDIA) cohort and its YALTA ancillary study who had either current asthma or never asthma at that evaluation. Obesity was defined as body mass index (BMI) >or=30 kg/m(2). Multivariable logistic regression analysis was performed with current asthma status as the dependent variable. RESULTS: Women, but not men, with current asthma had a lower mean unadjusted serum adiponectin concentration than those with never asthma (p<0.001; p for sex interaction <0.001). Similarly, current asthma was related to obesity only in women (OR 3.31, 95% CI 2.00 to 5.46, p for sex interaction = 0.004); this association was little affected by adjusting for serum adiponectin. The prevalence of current asthma in premenopausal women was reduced in the highest compared with the lowest tertile of serum adiponectin concentration (OR 0.46, 95% CI 0.26 to 0.84, p = 0.03), after adjusting for BMI. However, the interaction between serum adiponectin concentration and BMI category on current asthma status was not significant in premenopausal women or women overall. CONCLUSIONS: A high serum adiponectin concentration may protect against current asthma in premenopausal women but does not explain the association between asthma and adiposity. PMID- 18390630 TI - Isolated nocturnal desaturation in COPD: prevalence and impact on quality of life and sleep. AB - BACKGROUND AND AIMS: The clinical impact of nocturnal desaturation on health related quality of life (HRQL) and sleep in chronic obstructive pulmonary disease (COPD) has been little studied. The aim of this study was to evaluate the prevalence and clinical impact of nocturnal desaturation in a typical outpatient population with COPD. PATIENTS AND METHODS: Between 2002 and 2005, consecutive patients with COPD attending outpatient services at the study centre underwent resting oximetry if they were not on domiciliary oxygen therapy. If their resting saturations were less than 95%, overnight pulse oximetry was performed. Significant nocturnal desaturation was defined as spending more than 30% of at least one of two nights with a saturation of less than 90%. The Chronic Respiratory Questionnaire (CRQ) and Short Form 36 (SF36) were used to assess HRQL, and the Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Score (ESS) and Functional Outcomes of Sleep (FOSQ) questionnaires were used to assess sleep quality and daytime function. RESULTS: Of 1104 patients, 803 underwent resting oximetry and 79 had resting oxygen saturations of less than 95%. Of these, 59 agreed to undergo overnight oximetry (mean age 70 years, forced expiratory volume in 1 s 37.2% predicted, resting Po(2) on air 8.9 kPa). Significant nocturnal desaturation was seen in 29 (49.2%) of the 59 subjects. Assuming the less hypoxic patients do not have nocturnal desaturation, the prevalence of nocturnal desaturation in the whole clinic population could be estimated at 4.8%. There were no significant differences in CRQ, SF36, PSQI, ESS or FOSQ scores for desaturators compared with non-desaturators. CONCLUSION: Significant nocturnal desaturation was common in patients with COPD with resting saturations of less than 95%, but was estimated to have a prevalence of less than 5% in the whole outpatient population. Nocturnal desaturation was not associated with impairment of HRQL, sleep quality or daytime function. PMID- 18390631 TI - Effectiveness of third-generation chemotherapy on the survival of patients with advanced non-small cell lung cancer in Norway: a national study. AB - BACKGROUND: To investigate whether the introduction of modern third-generation chemotherapy was associated with survival benefits in a national population of patients with advanced non-small cell lung cancer (ANSCLC) and to explore geographical and temporary variations in the utilisation of chemotherapy. METHODS: All patients with ANSCLC in the Cancer Registry of Norway during 1994 2005 were included. Using sales of vinorelbine as an indicator for chemotherapy, annual county utilisation rates were calculated. Survival before and after the general introduction of vinorelbine and associations between survival and variations in utilisation in counties were investigated. In a subgroup, the predictors of having received chemotherapy were explored. RESULTS: Of 24 875 registered patients with lung cancer, 13 757 had ANSCLC. The annual utilisation of the indicator drug in Norway increased from 3.7 to 184.2 g (1998-2005). Median survival increased from 149 to176 days (p<0.001). The adjusted hazard ratio (HR) for a diagnosis after the introduction was 0.93 (95% CI 0.88 to 0.99). County utilisation rates of vinorelbine (increments of 100 mg/1000 inhabitants) were inversely associated with the risk of death (HR 0.84, 95% CI 0.73 to 0.98). County of residence predicted chemotherapy utilisation with odds ratios in the range 0.13 (95% CI 0.1 to 0.19) to 1.04 (95% CI 0.64 to 1.69), a county with traditionally high utilisation as reference. CONCLUSION: Utilisation of third generation chemotherapy was associated with slightly increased survival of patients with ANSCLC. Geographical and temporal differences in utilisation indicate variable quality of delivered care. PMID- 18390632 TI - Prediction equations for single breath diffusing capacity (Tlco) in a middle aged caucasian population. AB - BACKGROUND: There are many reference equations for the measurement of single breath carbon monoxide diffusing capacity of the lung (Tlco). However, the testing methodologies vary and there are no well documented studies that have developed reference equations for Tlco and alveolar volume (Va) in middle aged and older populations. AIMS: (1) Develop reference equations for Tlco in a middle aged population using the current American Thoracic Society/European Respiratory Society (ATS/ERS) guidelines; (2) compare the equations with those commonly used in laboratories around the world. METHODS: Healthy subjects (498 male and 474 female) aged 45-71 years were recruited as part of a larger epidemiological study. All participants completed a respiratory questionnaire and had spirometry and single breath Tlco (corrected for haemoglobin) measurements following ATS/ERS guidelines. RESULTS: Mean age was 58 years for males and 57 years for females. For males, factors that predicted Tlco were: height, age, agexheight interaction and being an ex-smoker. For females, factors that predicted Tlco were: height, age, weight and an agexheight interaction. CONCLUSION: We have described new prediction equations for Tlco in a middle aged population that require validation in other populations. PMID- 18390634 TI - Minimum clinically important improvement for the incremental shuttle walking test. AB - BACKGROUND: The incremental shuttle walking test (ISWT) is used to assess exercise capacity in patients with chronic obstructive pulmonary disease (COPD) and is employed as an outcome measure for pulmonary rehabilitation. This study was designed to establish the minimum clinically important difference for the ISWT. METHODS: 372 patients (205 men) performed an ISWT before and after a 7-week outpatient pulmonary rehabilitation programme. After completing the course, subjects were asked to identify, from a 5-point Likert scale, the perceived change in their exercise performance immediately upon completion of the ISWT. The scale ranged from "better" to "worse". RESULTS: The mean (SD) age was 69.4 (8.4) years, forced expiratory volume in 1 s (FEV(1)) 1.06 (0.53) l and FEV(1)/forced vital capacity (FVC) ratio 50.8 (18.1)%. The baseline shuttle walking test distance was 168.5 (114.6) m which increased to 234.7 (125.3) m after rehabilitation (mean difference 65.9 m (95% CI 58.9 to 72.9)). In subjects who felt their exercise tolerance was "slightly better" the mean improvement was 47.5 m (95% CI 38.6 to 56.5) compared with 78.7 m (95% CI 70.5 to 86.9) in those who reported that their exercise tolerance was "better" and 18.0 m (95% CI 4.5 to 31.5) in those who felt their exercise tolerance was "about the same". CONCLUSION: Two levels of improvement were identified. The minimum clinically important improvement for the ISWT is 47.5 m. In addition, patients were able to distinguish an additional benefit at 78.7 m. PMID- 18390633 TI - Analysis of cell cycle-related proteins in mediastinal lymph nodes of patients with N2-NSCLC obtained by EBUS-TBNA: relevance to chemotherapy response. AB - BACKGROUND: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is an accurate tool for lymph node staging of non-small cell lung cancer (NSCLC). Most patients with NSCLC require systemic chemotherapy during their treatment, with relatively poor responses. If the response to chemotherapy could be predicted, ideally at the time of the initial bronchoscopic examination, the therapeutic benefit could be maximised while limiting toxicity. A study was therefore undertaken to investigate the feasibility of EBUS-TBNA for obtaining tissue samples from mediastinal lymph nodes that can be used for immunohistochemical analysis, and to stratify patients with molecular-based pN2 NSCLC into chemo-responsive and chemoresistant subgroups who might benefit from tailoring of chemotherapy. METHODS: The expression of six cell cycle-related proteins (pRb, cyclin D1, p16(INK4A), p53, p21(Waf1), Ki-67) in mediastinal lymph node specimens obtained by EBUS-TBNA was investigated by immunohistochemistry in 36 patients with pN2-NSCLC. Their predictive role(s) in the response to platinum based chemotherapy was examined. RESULTS: Immunostaining was feasible in all studied specimens. Univariate analysis revealed that p53 and p21(Waf1) expressions were significantly related to the response to chemotherapy (p = 0.002 and p = 0.011, respectively). Multivariate logistic regression analysis revealed that only p53 overexpression was associated with a poor response to chemotherapy (p = 0.021). CONCLUSIONS: These results suggest that EBUS-TBNA is a feasible tool for obtaining mediastinal nodal tissue samples amenable for immunohistochemical analysis. Immunostaining of p53 in EBUS-TBNA-guided specimens may be useful in predicting the response to chemotherapy in patients with N2-NSCLC and helping in the selection of patients who might benefit from certain chemotherapeutic strategies. PMID- 18390635 TI - Continuous positive airway pressure improves vascular function in obstructive sleep apnoea/hypopnoea syndrome: a randomised controlled trial. AB - BACKGROUND: The obstructive sleep apnoea/hypopnoea syndrome (OSAHS) is associated with hypertension and increased cardiovascular risk, particularly when accompanied by marked nocturnal hypoxaemia. The mechanisms of these associations are unclear. We hypothesised that OSAHS combined with severe nocturnal hypoxaemia causes impaired vascular function that can be reversed by continuous positive airways pressure (CPAP) therapy. METHODS: We compared vascular function in two groups of patients with OSAHS: 27 with more than 20 4% desaturations/h (desaturator group) and 19 with no 4% and less than five 3% desaturations/h (non desaturator group). In a randomised, double blind, placebo controlled, crossover trial, the effect of 6 weeks of CPAP therapy on vascular function was determined in the desaturator group. In all studies, vascular function was assessed invasively by forearm venous occlusion plethysmography during intra-arterial infusion of endothelium dependent (acetylcholine 5-20 microg/min and substance P 2-8 pmol/min) and independent (sodium nitroprusside 2-8 microg/min) vasodilators. RESULTS: Compared with the non-desaturator group, patients with OSAHS and desaturations had reduced vasodilatation to all agonists (p = 0.007 for all). The apnoea/hypopnoea index and desaturation frequency were inversely related to peak vasodilatation with acetylcholine (r = -0.44, p = 0.002 and r = -0.43, p = 0.003) and sodium nitroprusside (r = -0.42, p = 0.009 and r = -0.37, p = 0.02). In comparison with placebo, CPAP therapy improved forearm blood flow to all vasodilators (p = 0.01). CONCLUSIONS: Patients with OSAHS and frequent nocturnal desaturations have impaired endothelial dependent and endothelial independent vasodilatation that is proportional to hypoxaemia and is improved by CPAP therapy. Impaired vascular function establishes an underlying mechanism for the adverse cardiovascular consequences of OSAHS. PMID- 18390636 TI - Versican and fibrillin-1 form a major hyaluronan-binding complex in the ciliary body. AB - PURPOSE: In this study, biochemistry, molecular biology, immunohistochemistry, and electron microscopy techniques were used to examine whether versican, which is known to bind fibrillin-1, interacts with fibrillin-1 in the ciliary body and vitreous, and whether the versican in this complex binds to hyaluronan. METHODS: The new polyclonal antibodies against the amino and carboxyl termini of versican were raised and characterized. The mRNA expression levels of versican and fibrillin-1 were analyzed by RT-PCR and real-time PCR, and their protein levels were evaluated by Western blot analysis and immunohistochemistry. Isolation of versican bound to fibrillin-1-containing microfibrils from ciliary bodies was performed by extraction studies. Slot-blot analyses and rotary shadowing electron microscopy were applied to identify versican associated with fibrillin-1 containing microfibrils after gel filtration chromatography and density gradient centrifugation. RESULTS: The newly prepared polyclonal antibodies recognized amino and carboxyl termini of chicken versican. Versican, principally V0 and V1, was found to be securely bound to fibrillin-1-containing microfibrils, forming a major hyaluronan-binding structure in the ciliary nonpigmented epithelium. In addition, Western blot analysis revealed two cleaved complexes, the carboxyl terminal end of versican bound to fibrillin microfibrils and the amino terminal end of versican bound to hyaluronan in the vitreous body. CONCLUSIONS: Fibrillin 1, versican, and hyaluronan form a unique complex in the ciliary nonpigmented epithelium, and two cleavage products of this complex were shown to exist in the vitreous body. This newly clarified fibrillin-versican-hyaluronan (FiVerHy) complex and its cleavage products may be indispensable for the physiological properties important to the ciliary body and vitreous. PMID- 18390637 TI - Autologous transplantation of RPE with partial-thickness choroid after mechanical debridement of Bruch membrane in the rabbit. AB - PURPOSE: An improved translocation technique for autologous retinal pigment epithelium (RPE) transplantation is presented. The graft consists of a sheet of a partial-thickness choroid with RPE attached. METHODS: Twenty-seven pigmented rabbits were used in this study. After mechanical debridement of Bruch membrane, partial-thickness RPE-choroid sheets were transplanted onto the subretinal space in 25 rabbits. The animals were examined by fundus photographs and fluorescein angiographs and were killed postoperatively at 1, 2, 4, 12, and 24 weeks. Eyecups containing the grafts were examined by light microscopy and immunohistochemistry. In addition, two partial-thickness RPE-choroid sheets were analyzed by transmission electron microscopy (TEM). RESULTS: TEM revealed that the partial thickness RPE-choroid graft consisted of retinal pigment epithelial cells, Bruch membrane, choriocapillaris, and ruptured middle vessels. The thickness of the graft was approximately 50 to 60 microm. Fluorescein angiography revealed neither fluorescein leakage nor staining in the graft at early or late phase. Light microscopy revealed that in 17 experiments in which the graft survived, the neural retina remained intact; however, in eight experiments with unsuccessful grafts, the neural retina degenerated. The surviving graft showed revascularization and monolayered retinal pigment epithelial cells. Furthermore, in sections in which the neural retina over the graft remained intact, all retinal pigment epithelial cells in the graft and rhodopsin in photoreceptor outer segments were positively labeled with anticellular retinaldehyde-binding protein antibodies and anti-opsin antibodies, respectively. CONCLUSIONS: A partial-thickness RPE-choroid graft showed improved integration with the host choroid and photoreceptors. This technique has the potential to be a treatment for age-related macular degeneration. PMID- 18390638 TI - In vivo three-dimensional imaging of neovascular age-related macular degeneration using optical frequency domain imaging at 1050 nm. AB - PURPOSE: To assess the application of optical frequency domain imaging (OFDI) at 1050 nm for the detection of choroidal neovascularization (CNV) in age-related macular degeneration (AMD) and its response to treatment. Three patients presenting with blurred vision and exudative AMD were imaged before and after anti-VEGF treatment with ranibizumab. METHODS: The patients were imaged with OFDI, a swept-source-based, high-speed optical coherence tomography (OCT) system developed at the Wellman Center for Photomedicine. A center wavelength of 1050 nm was used that has been demonstrated to provide better imaging of the deeper structures of the retina below the RPE, such as the choroidal vasculature. Three dimensional data sets were acquired in 2 to 4 seconds. RESULTS: En face images were compiled from cross-sectional OFDI data and correlated with color fundus photography (CF) and fluorescein angiograms (FAs). Cross-sectional images were coregistered with CF and FA to obtain depth-resolved information about CNV, CNV volume, retinal thickness, subretinal fluid volume and height of neurosensory detachment before and after treatment with ranibizumab. A band of reduced reflectivity below the RPE was identified in all three subjects that corresponded to areas of confirmed and suspected occult CNV on FA. After treatment, this band was reduced in volume in all patients. CONCLUSIONS: High-speed 3-D OFDI at 1050 nm is a promising technology for imaging the retina and choroid in neovascular AMD. The developed system at 1050 nm provides good contrast for occult (type 1) CNV and may have advantages compared with time domain and current state of the art spectral domain OCT systems (SD-OCT) at 850 nm. PMID- 18390639 TI - AMD-like lesions in the rat retina: a latent consequence of perinatal hemorrhage. AB - PURPOSE: Hemorrhaging is a commonplace event in the human retina around the time of birth. This study was conducted to examine the potential long-term sequelae of hemorrhaging in the eyes of rats that exhibited transient spontaneous microhemorrhages a few days after birth. METHODS: Retinas of Dark Agouti rats aged from day of birth to 2 years old were analyzed histologically, histochemically, and by immunocytochemistry. Fetal human retinas were also examined anatomically and histochemically for evidence of hemorrhages. RESULTS: Dark Agouti rats from our colony consistently exhibited spontaneous focal hemorrhages at the vitread surface of the retina between postnatal days 3 and 6. Erythrocytes were subsequently cleared by macrophages, which accumulated hemosiderin. These macrophages remained in focal patches in the inner retina for the duration of the study. For at least 6 months after the initial transient hemorrhages, the retinas exhibited no overt histologic damage. At approximately 8 to 9 months, photoreceptor degenerative changes were apparent in spatial register with the patches of macrophages in the inner retina. Additional events such as breakdown of Bruch's membrane, glial remodeling, neovascularization, ingress of RPE cells into the retina and accumulation of drusen-like autofluorescent structures were also observed in topographic register with macrophage-laden patches in aged animals. CONCLUSIONS: Microhemorrhages in the retina may initiate the formation of focal lesions, months or years after the initial insult. The lesions exhibit key features of AMD. These animals may represent a useful model for studying the potential basis of the pathogenesis of AMD. PMID- 18390640 TI - Effect of intravitreal administration of somatostatin and sst2 analogs on AMPA induced neurotoxicity in rat retina. AB - PURPOSE: The aim of the present study was to use an in vivo model of retinal excitotoxicity to investigate the neuroprotective effect of somatostatin (SRIF) ergic agents. METHODS: Adult Sprague-Dawley rats (weight range, 250-300 g) intravitreally received (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid hydrobromide (AMPA; 21, 42, 84 nmol/eye) or PBS (50 mM). Time-dependent responses were examined in animals that received AMPA (42 nmol/eye). Animals received AMPA (42 nmol) alone or in combination with SRIF (10(-5), 10(-4) M) or the sst-selective ligands lanreotide (sst(2),10(-5),10(-4) M), L-779976 (sst(2,)10(-6),10(-5), 10(-4) M), L-797591 (sst(1),10(-4) M), and L-803087 (sst(4),10(-4) M). Immunohistochemistry and TUNEL studies were used to examine retinal cell loss and protection. Immunochemistry, Western blot analysis, and radioimmunoassay assessed the viability of sst(2A) receptors and SRIF levels, respectively, in control and AMPA-treated tissue. RESULTS: AMPA (42 nmol) treatment resulted in total and major loss of ChAT and bNOS immunoreactivity, respectively, 24 hours after its administration. This loss was sustained up to 30 days for ChAT- and 8 days for bNOS-expressing amacrine cells. SRIF and the sst(2) receptors were not affected by AMPA. SRIF and the sst(2) analogs protected the retina from the AMPA insult in a dose-dependent manner, whereas activation of the sst(1) and sst(4) subtypes had no effect. TUNEL staining confirmed AMPA-induced retinal ischemia and L-779976 neuroprotection. CONCLUSIONS: These results demonstrate for the first time that SRIF and the sst(2) analogs, administered intravitreally, protect the retina from excitotoxicity. Further studies are essential to ascertain the therapeutic relevance of these results. PMID- 18390641 TI - Task-induced stress and motivation decrease foveation-period durations in infantile nystagmus syndrome. AB - PURPOSE: To investigate the effect of visual demand, task-related physiological stress, and motivation on the nystagmus waveform of 19 subjects with infantile nystagmus syndrome (INS). METHODS: Subjects viewed a Landolt C of varying orientation and size, and indicated its orientation via arrow keys on a keyboard. Mental arithmetic was performed in conjunction with the visual task. Subjects then underwent a reward-penalty paradigm. Eye movements and heart rates were recorded during all experiments. RESULTS: Task-related physiological stress and motivation were reflected in an increase in heart rate and led to an increase in the amplitude, frequency, and intensity of the nystagmus waveform and a decrease in foveation-period durations. Changes in heart rate did not correlate with changes in waveform parameters for all experiments. CONCLUSIONS: The results show, for the first time, the negative impact of task-induced stress and/or motivation on the characteristics of INS. This finding has important implications for individuals with INS, because stress may arise in everyday situations, such as driving or when undertaking an examination. PMID- 18390642 TI - Increased role of peripheral vision in self-induced motion in patients with age related macular degeneration. AB - PURPOSE: The contribution of peripheral vision in inducing self-motion (vection) was investigated in people with bilateral age-related macular degeneration (AMD). METHODS: Eleven patients with bilateral AMD and dense central scotomas with no islands of functional central retina and 12 age-matched control subjects were exposed to random-dot patterns projected on a large screen. The dots either moved from left to right, inducing linear vection, or rotated about the roll axis, inducing roll vection. Latency, total vection time, and objective and subjective measures of tilt were recorded. RESULTS: The patients with AMD experienced shorter latencies than did the age-matched control participants, but the total vection time in both conditions and tilt during roll vection were the same in both groups. There was a positive correlation between objective tilt and subjective measures of tilt in the AMD, but not in the age-matched control group. There was a negative relationship between absolute scotoma size and latency. CONCLUSIONS: Two main conclusions were drawn. First, the role of peripheral vision in inducing vection is enhanced in people with bilateral central vision loss. Second, people with bilateral AMD adapt successfully to a moving environment (they do not experience vection longer, nor do they tilt more) and are more aware of their postural position than are age-matched control subjects. PMID- 18390643 TI - Prevalence and associated risk factors of age-related macular degeneration in an elderly Chinese population in Taiwan: the Shihpai Eye Study. AB - PURPOSE: To assess the prevalence and associated risk factors of age-related macular degeneration (AMD) in an elderly Chinese population in Taiwan. METHODS: The Shihpai Eye Study was a survey of vision and ocular disease in an elderly Chinese population 65 years of age or older residing in Shihpai, Taipei, Taiwan. Of 2045 elderly residents randomly sampled from the household registration databank, 1361 (66.6%) underwent a detailed ophthalmic examination that included fundus color slides by fundus camera after pupil dilatation. Photographs were graded according to the Wisconsin Age-Related Maculopathy Grading System. RESULTS: Fundus photographs were available for 1105 (54.0% in the eligible, 81.2% in the ocular examined) participants. The 47 (4.3%) participants who had ungradable fundus images were older and had more lens opacity. Of the 1058 gradable photographs, the prevalence of early AMD was 9.2% (95% confidence interval [CI], 7.8-10.8); of late AMD, 1.9% (95% CI, 1.3-2.7); of soft drusen, 42.2% (95% CI, 39.7-44.8); of soft indistinct drusen, 4.1% (95% CI, 3.1-5.2); and of any pigmentary change, 8.6% (95% CI, 7.2-10.2). Age was the most significant factor associated with both early and late AMD. The prevalence of early AMD rose from 5.0% in the 65- to 69-year age group to 24.4% in those 80 years of age and older; and for late AMD, from 1.0% to 9.0%. Those who currently drank alcohol had a lower rate of early AMD than did the nondrinker (adjusted odd ratio 0.32, 95% CI: 0.11-0.93, P = 0.037). CONCLUSIONS: AMD is a common eye disease in the elderly Chinese people in Taiwan. The adjusted prevalence rate of exudative AMD is comparable to that in the Chinese people in the Multiethnic Study of Atherosclerosis (MESA) in the United States but is higher than in the Chinese people in the Beijing study in China. Further studies are needed to clarify the incidence and associated risk factors. PMID- 18390644 TI - Increasing incidence of ophthalmic lymphoma in Denmark from 1980 to 2005. AB - PURPOSE: To evaluate patient characteristics and incidence of ophthalmic lymphoma in Denmark during the period 1980 to 2005. METHODS: All patients in Denmark with a diagnosis of ophthalmic lymphoma during the period 1980 to 2005 were retrieved from three different population-based registries. Specimens from all patients were collected and reclassified according to the World Health Organization (WHO) classification system. Incidence rates were calculated by using Poisson regression models. RESULTS: A total of 228 patients with a histologically verified diagnosis of ophthalmic lymphoma were included. There was an equal distribution of males and females. The most frequent lymphoma subtype was extranodal marginal zone B-cell lymphoma (MALT [mucosa-associated lymphoid tissue] lymphoma, 55.5%) and most cases were located in the orbit (56.8%). High grade lymphoma subtypes were found more frequently in males than in females. Incidence rates were highly dependent on the patient's age. For all ages, a statistically significant annual average increase of 3.4% during the 26-year period was found. This increase was primarily due to a rise in the incidence of MALT lymphoma. CONCLUSIONS: In the Danish population ophthalmic lymphoma consists primarily of orbital MALT lymphoma. Although it is a rare disease in mostly elderly patients, the incidence of ophthalmic lymphoma is increasing at a rapid pace. PMID- 18390645 TI - Conjunctival and corneal pneumatic sensitivity is associated with signs and symptoms of ocular dryness. AB - PURPOSE: To investigate the relationships of dry eye symptoms and corneal and conjunctival sensitivity to pneumatic stimulation, tear film stability, and clinical ocular surface characteristics in symptomatic and asymptomatic subjects. METHODS: Ninety-seven subjects were enrolled and grouped by a questionnaire-based single score for symptoms of ocular dryness (none to trace, non-dry group; mild to severe, symptomatic group); 43 were symptomatic and 54 were non-dry. Corneal (K) and conjunctival (C) sensitivities were measured with a computer-controlled Belmonte pneumatic (room temperature) stimulus. Symptoms were assessed according to the Ocular Surface Disease Index (OSDI). Ocular surface staining with fluorescein (FL) and lissamine green (LG), noninvasive tear film break-up time (NIBUT), and the phenol red thread test (PRT) were assessed. RESULTS: The symptomatic group showed lower K and C thresholds (P < 0.01), greater corneal FL and conjunctival LG staining, and shorter NIBUT than did the non-dry eye group (all others P < 0.05). The OSDI scores were higher in the symptomatic group (P < 0.001). K and C thresholds and NIBUT were inversely correlated with the OSDI and corneal and conjunctival staining (all P < 0.05). The K and C threshold and NIBUT (all P < 0.01) correlated positively. Step-wise multiple regression analysis showed that ocular surface sensitivity and NIBUT were significant predictors of the OSDI. CONCLUSIONS: Ocular irritation assessed with the OSDI is associated with ocular surface hyperesthesia to cooling, corneal epitheliopathy, and tear film instability. Although cause and effect are unclear, the analysis showed that altered corneal and conjunctival sensory processing and tear film attributes are essential aspects of what characterizes dry eye. PMID- 18390646 TI - Comparison of aspects of smoking among the four histological types of lung cancer. AB - BACKGROUND: The magnitude of the link between cigarette smoking and lung cancer may vary by histological type. METHODS: We used polytomous logistic regression to evaluate whether aspects of smoking have different effects across four histological types in the Nurses' Health Study. RESULTS: From 1976 to 2002, we identified 1062 cases of lung cancer: squamous cell (n = 201), small cell (n = 236), adenocarcinoma (n = 543) and large cell carcinoma (n = 82), among 65 560 current or former smokers. Risk reduction after quitting ranged from an 8% reduction (relative risk (RR): 0.92, 95% CI 0.91 to 0.94) to a 17% reduction (RR: 0.83, 95% CI 0.80 to 0.86) per year for adenocarcinoma and small cell carcinoma, respectively, with a 9% reduction observed for large cell carcinoma and an 11% reduction observed for squamous cell carcinoma. The association of age at smoking initiation and former cigarette smoking was similar across types, while the association of smoking duration differed. The risk of adenocarcinoma increased by 6% per year of smoking, compared to 7% for large cell, 10% for squamous cell and 12% for small cell. The 6% difference between adenocarcinoma and small cell carcinoma is equivalent to a 3.2 to 9.7-fold increase in risk for 20 years of smoking. CONCLUSIONS: The effects of the number of cigarettes smoked per day and years since quitting smoking are different across the major types of lung cancer, which are fully appreciated at long durations of smoking and smoking cessation. Smoking prevention and cessation should continue to be the focus of public health efforts to reduce lung cancer incidence and mortality. PMID- 18390647 TI - The nuclear migration protein NUDF/LIS1 forms a complex with NUDC and BNFA at spindle pole bodies. AB - Nuclear migration depends on microtubules, the dynein motor complex, and regulatory components like LIS1 and NUDC. We sought to identify new binding partners of the fungal LIS1 homolog NUDF to clarify its function in dynein regulation. We therefore analyzed the association between NUDF and NUDC in Aspergillus nidulans. NUDF and NUDC directly interacted in yeast two-hybrid experiments via NUDF's WD40 domain. NUDC-green fluorescent protein (NUDC-GFP) was localized to immobile dots in the cytoplasm and at the hyphal cortex, some of which were spindle pole bodies (SPBs). We showed by bimolecular fluorescence complementation microscopy that NUDC directly interacted with NUDF at SPBs at different stages of the cell cycle. Applying tandem affinity purification, we isolated the NUDF-associated protein BNFA (for binding to NUDF). BNFA was dispensable for growth and for nuclear migration. GFP-BNFA fusions localized to SPBs at different stages of the cell cycle. This localization depended on NUDF, since the loss of NUDF resulted in the cytoplasmic accumulation of BNFA. BNFA did not bind to NUDC in a yeast two-hybrid assay. These results show that the conserved NUDF and NUDC proteins play a concerted role at SPBs at different stages of the cell cycle and that NUDF recruits additional proteins specifically to the dynein complex at SPBs. PMID- 18390648 TI - Molecular signals in the trafficking of Toxoplasma gondii protein MIC3 to the micronemes. AB - The protozoan parasite Toxoplasma gondii is equipped with a sophisticated secretory apparatus, including three distinct exocytic organelles, named micronemes, rhoptries, and dense granules. We have dissected the requirements for targeting the microneme protein MIC3, a key component of T. gondii infection. We have shown that MIC3 is processed in a post-Golgi compartment and that the MIC3 propeptide and epidermal growth factor (EGF) modules contain microneme-targeting information. The minimal requirement for microneme delivery is defined by the propeptide plus any one of the three EGF domains. We have demonstrated that the cleavage of the propeptide, the dimerization of MIC3, and the chitin binding-like sequence, which are crucial for host cell binding and virulence, are dispensable for proper targeting. Finally, we have shown that part of MIC3 is withheld in the secretory pathway in a cell cycle-dependent manner. PMID- 18390649 TI - Genomewide location analysis of Candida albicans Upc2p, a regulator of sterol metabolism and azole drug resistance. AB - Upc2p, a transcription factor of the zinc cluster family, is an important regulator of sterol biosynthesis and azole drug resistance in Candida albicans. To better understand Upc2p function in C. albicans, we used genomewide location profiling to identify the transcriptional targets of Upc2p in vivo. A triple hemagglutinin epitope, introduced at the C terminus of Upc2p, conferred a gain-of function effect on the fusion protein. Location profiling identified 202 bound promoters (P < 0.05). Overrepresented functional groups of genes whose promoters were bound by Upc2p included 12 genes involved in ergosterol biosynthesis (NCP1, ERG11, ERG2, and others), 18 genes encoding ribosomal subunits (RPS30, RPL32, RPL12, and others), 3 genes encoding drug transporters (CDR1, MDR1, and YOR1), 4 genes encoding transcription factors (INO2, ACE2, SUT1, and UPC2), and 6 genes involved in sulfur amino acid metabolism (MET6, SAM2, SAH1, and others). Bioinformatic analyses suggested that Upc2p binds to the DNA motif 5'-VNCGBDTR that includes the previously characterized Upc2p binding site 5'-TCGTATA. Northern blot analysis showed that increased binding correlates with increased expression for the analyzed Upc2p targets (ERG11, MDR1, CDR1, YOR1, SUT1, SMF12, and CBP1). The analysis of ERG11, MDR1, and CDR1 transcripts in wild-type and upc2Delta/upc2Delta strains grown under Upc2p-activating conditions (lovastatin treatment and hypoxia) showed that Upc2p regulates its targets in a complex manner, acting as an activator or as a repressor depending upon the target and the activating condition. Taken together, our results indicate that Upc2p is a key regulator of ergosterol metabolism. They also suggest that Upc2p may contribute to azole resistance by regulating the expression of drug efflux pump encoding genes in addition to ergosterol biosynthesis genes. PMID- 18390650 TI - Mutations in ampG or ampD affect peptidoglycan fragment release from Neisseria gonorrhoeae. AB - Neisseria gonorrhoeae releases peptidoglycan fragments during growth. The majority of the fragments released are peptidoglycan monomers, molecules known to increase pathogenesis through the induction of proinflammatory cytokines and responsible for the killing of ciliated epithelial cells. In other gram-negative bacteria such as Escherichia coli, these peptidoglycan fragments are efficiently degraded and recycled. Peptidoglycan fragments enter the cytoplasm from the periplasm via the AmpG permease. The amidase AmpD degrades peptidoglycan monomers by removing the disaccharide from the peptide. The disaccharide and the peptide are further degraded and are then used for new peptidoglycan synthesis or general metabolism. We examined the possibility that peptidoglycan fragment release by N. gonorrhoeae results from defects in peptidoglycan recycling. The deletion of ampG caused a large increase in peptidoglycan monomer release. Analysis of cytoplasmic material showed peptidoglycan fragments as recycling intermediates in the wild type strain but absent from the ampG mutant. An ampD deletion reduced the release of all peptidoglycan fragments and nearly eliminated the release of free disaccharide. The ampD mutant also showed a large buildup of peptidoglycan monomers in the cytoplasm. The introduction of an ampG mutation in the ampD background restored peptidoglycan fragment release, indicating that events in the cytoplasm (metabolic or transcriptional regulation) affect peptidoglycan fragment release. The ampD mutant showed increased metabolism of exogenously added free disaccharide derived from peptidoglycan. These results demonstrate that N. gonorrhoeae has an active peptidoglycan recycling pathway and can regulate peptidoglycan fragment metabolism, dependent on the intracellular concentration of peptidoglycan fragments. PMID- 18390651 TI - Norepinephrine mediates acquisition of transferrin-iron in Bordetella bronchiseptica. AB - Previous research demonstrated that the sympathoadrenal catecholamine norepinephrine could promote the growth of Bordetella bronchiseptica in iron restricted medium containing serum. In this study, norepinephrine was demonstrated to stimulate growth of this organism in the presence of partially iron-saturated transferrin but not lactoferrin. Although norepinephrine is known to induce transcription of the Bordetella bfeA enterobactin catechol xenosiderophore receptor gene, neither a bfeA mutant nor a bfeR regulator mutant was defective in growth responsiveness to norepinephrine. However, growth of a tonB mutant strain was not enhanced by norepinephrine, indicating that the response to this catecholamine was the result of high-affinity outer membrane transport. The B. bronchiseptica genome encodes a total of 19 known and predicted iron transport receptor genes, none of which, when mutated individually, were found to confer a defect in norepinephrine-mediated growth stimulation in the presence of transferrin. Labeling experiments demonstrated a TonB-dependent increase in cell-associated iron levels when bacteria grown in the presence of (55)Fe-transferrin were exposed to norepinephrine. In addition, TonB was required for maximum levels of cell-associated norepinephrine. Together, these results demonstrate that norepinephrine facilitates B. bronchiseptica iron acquisition from the iron carrier protein transferrin and this process may represent a mechanism by which some bacterial pathogens obtain this essential nutrient in the host environment. PMID- 18390652 TI - Identification of Escherichia coli YgaF as an L-2-hydroxyglutarate oxidase. AB - YgaF, a protein of previously unknown function in Escherichia coli, was shown to possess noncovalently bound flavin adenine dinucleotide and to exhibit L-2 hydroxyglutarate oxidase activity. The inability of anaerobic, reduced enzyme to reverse the reaction by reducing the product alpha-ketoglutaric acid is explained by the very high reduction potential (+19 mV) of the bound cofactor. The likely role of this enzyme in the cell is to recover alpha-ketoglutarate mistakenly reduced by other enzymes or formed during growth on propionate. On the basis of the identified function, we propose that this gene be renamed lhgO. PMID- 18390653 TI - EspA, an orphan hybrid histidine protein kinase, regulates the timing of expression of key developmental proteins of Myxococcus xanthus. AB - Myxococcus xanthus undergoes a complex starvation-induced developmental program that results in cells forming multicellular fruiting bodies by aggregating into mounds and then differentiating into spores. This developmental program requires at least 72 h and is mediated by a temporal cascade of gene regulators in response to intra- and extracellular signals. espA mutants, encoding an orphan hybrid histidine kinase, alter the timing of this developmental program, greatly accelerating developmental progression. Here, we characterized EspA and demonstrated that it autophosphorylates in vitro on the conserved histidine residue and then transfers the phosphoryl group to the conserved aspartate residue in the associated receiver domain. The conserved histidine and aspartate residues were both required for EspA function in vivo. Analysis of developmental gene expression and protein accumulation in espA mutants indicated that the expression of the A-signal-dependent spi gene was not affected but that the MrpC transcriptional regulator accumulated earlier, resulting in earlier expression of its target, the FruA transcriptional regulator. Early expression of FruA correlated with acceleration of both the aggregation and sporulation branches of the developmental program, as monitored by early methylation of the FrzCD chemosensory receptor and early expression of the sporulation-specific dev and Mxan_3227 (Omega7536) genes. These results show that EspA plays a key role in the timing of expression of genes necessary for progression of cells through the developmental program. PMID- 18390654 TI - Pseudomonas aeruginosa twitching motility-mediated chemotaxis towards phospholipids and fatty acids: specificity and metabolic requirements. AB - Pseudomonas aeruginosa demonstrates type IV pilus-mediated directional twitching motility up a gradient of phosphatidylethanolamine (PE). Only one of four extracellular phospholipases C of P. aeruginosa (i.e., PlcB), while not required for twitching motility per se, is required for twitching-mediated migration up a gradient of PE or phosphatidylcholine. Whether other lipid metabolism genes are associated with this behavior was assessed by analysis of transcription during twitching up a PE gradient in comparison to transcription during twitching in the absence of any externally applied phospholipid. Data support the hypothesis that PE is further degraded and that the long-chain fatty acid (LCFA) moieties of PE are completely metabolized via beta-oxidation and the glyoxylate shunt. It was discovered that P. aeruginosa exhibits twitching-mediated chemotaxis toward unsaturated LCFAs (e.g., oleic acid), but not saturated LCFAs (e.g., stearic acid) of corresponding lengths. Analysis of mutants that are deficient in glyoxylate shunt enzymes, specifically isocitrate lyase (DeltaaceA) and malate synthase (DeltaaceB), suggested that the complete metabolism of LCFAs through this pathway was required for the migration of P. aeruginosa up a gradient of PE or unsaturated LCFAs. At this point, our data suggested that this process should be classified as energy taxis. However, further evaluation of the ability of the DeltaaceA and DeltaaceB mutants to migrate up a gradient of PE or unsaturated LCFAs in the presence of an alternative energy source clearly indicated that metabolism of LCFAs for energy is not required for chemotaxis toward these compounds. PMID- 18390655 TI - Photoregulation of cellular morphology during complementary chromatic adaptation requires sensor-kinase-class protein RcaE in Fremyella diplosiphon. AB - We used wild-type UTEX481; SF33, a shortened-filament mutant strain that shows normal complementary chromatic adaptation pigmentation responses; and FdBk14, an RcaE-deficient strain that lacks light-dependent pigmentation responses, to investigate the molecular basis of the photoregulation of cellular morphology in the cyanobacterium Fremyella diplosiphon. Detailed microscopic and biochemical analyses indicate that RcaE is required for the photoregulation of cell and filament morphologies of F. diplosiphon in response to red and green light. PMID- 18390656 TI - Growth of Escherichia coli: significance of peptidoglycan degradation during elongation and septation. AB - We have found a striking difference between the modes of action of amdinocillin (mecillinam) and compound A22, both of which inhibit cell elongation. This was made possible by employment of a new method using an Escherichia coli peptidoglycan (PG)-recycling mutant, lacking ampD, to analyze PG degradation during cell elongation and septation. Using this method, we have found that A22, which is known to prevent MreB function, strongly inhibited PG synthesis during elongation. In contrast, treatment of elongating cells with amdinocillin, which inhibits penicillin-binding protein 2 (PBP2), allowed PG glycan synthesis to proceed at a nearly normal rate with concomitant rapid degradation of the new glycan strands. By treating cells with A22 to inhibit sidewall synthesis, the method could also be applied to study septum synthesis. To our surprise, over 30% of newly synthesized septal PG was degraded during septation. Thus, excess PG sufficient to form at least one additional pole was being synthesized and rapidly degraded during septation. We propose that during cell division, rapid removal of the excess PG serves to separate the new poles of the daughter cells. We have also employed this new method to demonstrate that PBP2 and RodA are required for the synthesis of glycan strands during elongation and that the periplasmic amidases that aid in cell separation are minor players, cleaving only one-sixth of the PG that is turned over by the lytic transglycosylases. PMID- 18390657 TI - Opr86 is essential for viability and is a potential candidate for a protective antigen against biofilm formation by Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is an opportunistic bacterial pathogen that is one of the most refractory to therapy when it forms biofilms in the airways of cystic fibrosis patients. To date, studies regarding the production of an immunogenic and protective antigen to inhibit biofilm formation by P. aeruginosa have been superficial. The previously uncharacterized outer membrane protein (OMP) Opr86 (PA3648) of P. aeruginosa is a member of the Omp85 family, of which homologs have been found in all gram-negative bacteria. Here we verify the availability of Opr86 as a protective antigen to inhibit biofilm formation by P. aeruginosa PAO1 and several other isolates. A mutant was constructed in which Opr86 expression could be switched on or off through a tac promoter-controlled opr86 gene. The result, consistent with previous Omp85 studies, showed that Opr86 is essential for viability and plays a role in OMP assembly. Depletion of Opr86 resulted in streptococci-like morphological changes and liberation of excess membrane vesicles. A polyclonal antibody against Opr86 which showed reactivity to PAO1 cells was obtained. The antibody inhibited biofilm formation by PAO1 and the other clinical strains tested. Closer examination of early attachment revealed that cells treated with the antibody were unable to attach to the surface. Our data suggest that Opr86 is a critical OMP and a potential candidate as a protective antigen against biofilm formation by P. aeruginosa. PMID- 18390658 TI - Insight from TonB hybrid proteins into the mechanism of iron transport through the outer membrane. AB - We created hybrid proteins to study the functions of TonB. We first fused the portion of Escherichia coli tonB that encodes the C-terminal 69 amino acids (amino acids 170 to 239) of TonB downstream from E. coli malE (MalE-TonB69C). Production of MalE-TonB69C in tonB(+) bacteria inhibited siderophore transport. After overexpression and purification of the fusion protein on an amylose column, we proteolytically released the TonB C terminus and characterized it. Fluorescence spectra positioned its sole tryptophan (W213) in a weakly polar site in the protein interior, shielded from quenchers. Affinity chromatography showed the binding of the TonB C-domain to other proteins: immobilized TonB-dependent (FepA and colicin B) and TonB-independent (FepADelta3-17, OmpA, and lysozyme) proteins adsorbed MalE-TonB69C, revealing a general affinity of the C terminus for other proteins. Additional constructions fused full-length TonB upstream or downstream of green fluorescent protein (GFP). TonB-GFP constructs had partial functionality but no fluorescence; GFP-TonB fusion proteins were functional and fluorescent. The activity of the latter constructs, which localized GFP in the cytoplasm and TonB in the cell envelope, indicate that the TonB N terminus remains in the inner membrane during its biological function. Finally, sequence analyses revealed homology in the TonB C terminus to E. coli YcfS, a proline-rich protein that contains the lysin (LysM) peptidoglycan-binding motif. LysM structural mimicry occurs in two positions of the dimeric TonB C-domain, and experiments confirmed that it physically binds to the murein sacculus. Together, these findings infer that the TonB N terminus remains associated with the inner membrane, while the downstream region bridges the cell envelope from the affinity of the C terminus for peptidoglycan. This architecture suggests a membrane surveillance model of action, in which TonB finds occupied receptor proteins by surveying the underside of peptidoglycan-associated outer membrane proteins. PMID- 18390659 TI - Characterization of a novel methanol dehydrogenase in representatives of Burkholderiales: implications for environmental detection of methylotrophy and evidence for convergent evolution. AB - Some members of Burkholderiales are able to grow on methanol but lack the genes (mxaFI) responsible for the well-characterized two-subunit pyrroloquinoline quinone-dependent quinoprotein methanol dehydrogenase that is widespread in methylotrophic Proteobacteria. Here, we characterized novel, mono-subunit enzymes responsible for methanol oxidation in four strains, Methyloversatilis universalis FAM5, Methylibium petroleiphilum PM1, and unclassified Burkholderiales strains RZ18-153 and FAM1. The enzyme from M. universalis FAM5 was partially purified and subjected to matrix-assisted laser desorption ionization-time of fight peptide mass fingerprinting. The resulting peptide spectrum was used to identify a gene candidate in the genome of M. petroleiphilum PM1 (mdh2) predicted to encode a type I alcohol dehydrogenase related to the characterized methanol dehydrogenase large subunits but at less than 35% amino acid identity. Homologs of mdh2 were amplified from M. universalis FAM5 and strains RZ18-153 and FAM1, and mutants lacking mdh2 were generated in three of the organisms. These mutants lost their ability to grow on methanol and ethanol, demonstrating that mdh2 is responsible for oxidation of both substrates. Our findings have implications for environmental detection of methylotrophy and indicate that this ability is widespread beyond populations possessing mxaF, the gene traditionally used as a genetic marker for environmental detection of methanol-oxidizing capability. Our findings also have implications for understanding the evolution of methanol oxidation, suggesting a convergence toward the enzymatic function for methanol oxidation in MxaF and Mdh2-type proteins. PMID- 18390660 TI - Cyclic AMP in mycobacteria: characterization and functional role of the Rv1647 ortholog in Mycobacterium smegmatis. AB - Mycobacterial genomes are endowed with many eukaryote-like nucleotide cyclase genes encoding proteins that can synthesize 3',5'-cyclic AMP (cAMP). However, the roles of cAMP and the need for such redundancy in terms of adenylyl cyclase genes remain unknown. We measured cAMP levels in Mycobacterium smegmatis during growth and under various stress conditions and report the first biochemical and functional characterization of the MSMEG_3780 adenylyl cyclase, whose orthologs in Mycobacterium tuberculosis (Rv1647) and Mycobacterium leprae (ML1399) have been recently characterized in vitro. MSMEG_3780 was important for producing cAMP levels in the logarithmic phase of growth, since the DeltaMSMEG_3780 strain showed lower intracellular cAMP levels at this stage of growth. cAMP levels decreased in wild-type M. smegmatis under conditions of acid stress but not in the DeltaMSMEG_3780 strain. This was correlated with a reduction in MSMEG_3780 promoter activity, indicating that the effect of the reduction in cAMP levels on acid stress was caused by a decrease in the transcription of MSMEG_3780. Complementation of the DeltaMSMEG_3780 strain with the genomic integration of MSMEG_3780 or the Rv1647 gene could restore cAMP levels during logarithmic growth. The Rv1647 promoter was also acid sensitive, emphasizing the biochemical and functional similarities in these two adenylyl cyclases. This study therefore represents the first detailed biochemical and functional analysis of an adenylyl cyclase that is important for maintaining cAMP levels in mycobacteria and underscores the subtle roles that these genes may play in the physiology of the organism. PMID- 18390661 TI - Insertion hot spot for horizontally acquired DNA within a bidirectional small-RNA locus in Salmonella enterica. AB - In Escherichia coli and Salmonella enterica, RyeA and RyeB RNAs are encoded on opposite DNA strands at the same locus. We present evidence indicating that the last 23 bp of the ryeB gene, corresponding to an internal portion of the ryeA gene, served repeatedly as the integration site for exogenous DNA during Salmonella evolution and still act as an attachment site for present-day bacteriophages. Interestingly, ryeA sequence and expression are modified upon lysogenization. PMID- 18390662 TI - The Methanothermobacter thermautotrophicus Cdc6-2 protein, the putative helicase loader, dissociates the minichromosome maintenance helicase. AB - The Cdc6-1 and -2 proteins from the archaeon Methanothermobacter thermautotrophicus were previously shown to bind the minichromosome maintenance (MCM) helicase. It is shown here that Cdc6-2 protein dissociates the MCM complex. This observation supports the hypothesis that the Cdc6-2 protein functions as a helicase loader. PMID- 18390663 TI - Differential transcriptional analysis of the cyanobacterium Cyanothece sp. strain ATCC 51142 during light-dark and continuous-light growth. AB - We analyzed the metabolic rhythms and differential gene expression in the unicellular, diazotrophic cyanobacterium Cyanothece sp. strain ATCC 51142 under N(2)-fixing conditions after a shift from normal 12-h light-12-h dark cycles to continuous light. We found that the mRNA levels of approximately 10% of the genes in the genome demonstrated circadian behavior during growth in free-running (continuous light) conditions. The genes for N(2) fixation displayed a strong circadian behavior, whereas photosynthesis and respiration genes were not as tightly regulated. One of our main objectives was to determine the strategies used by these cells to perform N(2) fixation under normal day-night conditions, as well as under the greater stress caused by continuous light. We determined that N(2) fixation cycled in continuous light but with a lower N(2) fixation activity. Glycogen degradation, respiration, and photosynthesis were also lower; nonetheless, O(2) evolution was about 50% of the normal peak. We also demonstrated that nifH (encoding the nitrogenase Fe protein), nifB, and nifX were strongly induced in continuous light; this is consistent with the role of these proteins during the assembly of the enzyme complex and suggested that the decreased N(2) fixation activity was due to protein-level regulation or inhibition. Many soluble electron carriers (e.g., ferredoxins), as well as redox carriers (e.g., thioredoxin and glutathione), were strongly induced during N(2) fixation in continuous light. We suggest that these carriers are required to enhance cyclic electron transport and phosphorylation for energy production and to maintain appropriate redox levels in the presence of elevated O(2), respectively. PMID- 18390664 TI - Functional and phylogenetic characterization of Vaginolysin, the human-specific cytolysin from Gardnerella vaginalis. AB - Pore-forming toxins are essential to the virulence of a wide variety of pathogenic bacteria. Gardnerella vaginalis is a bacterial species associated with bacterial vaginosis (BV) and its significant adverse sequelae, including preterm birth and acquisition of human immunodeficiency virus. G. vaginalis makes a protein toxin that generates host immune responses and has been hypothesized to be involved in the pathogenesis of BV. We demonstrate that G. vaginalis produces a toxin (vaginolysin [VLY]) that is a member of the cholesterol-dependent cytolysin (CDC) family, most closely related to intermedilysin from Streptococcus intermedius. Consistent with this predicted relationship, VLY lyses target cells in a species-specific manner, dependent upon the complement regulatory molecule CD59. In addition to causing erythrocyte lysis, VLY activates the conserved epithelial p38 mitogen-activated protein kinase pathway and induces interleukin-8 production by human epithelial cells. Transfection of human CD59 into nonsusceptible cells renders them sensitive to VLY-mediated lysis. In addition, a single amino acid substitution in the VLY undecapeptide [VLY(P480W)] generates a toxoid that does not form pores, and introduction of the analogous proline residue into another CDC, pneumolysin, significantly decreases its cytolytic activity. Further investigation of the mechanism of action of VLY may improve understanding of the functions of the CDC family as well as diagnosis and therapy for BV. PMID- 18390665 TI - Glycine betaine uptake by the ProXVWZ ABC transporter contributes to the ability of Mycobacterium tuberculosis to initiate growth in human macrophages. AB - Mycobacterium tuberculosis maintains a large genetic capacity necessary for growth in different environments during infection and survival upon aerosol transmission to new hosts. Screening for bacterial RNAs produced in response to host interactions produced candidate lists where we noted proXVWZ, annotated as encoding a putative glycine betaine or proline transporter. As high surface-to volume ratios make bacterial cells particularly vulnerable to changes in water availability, we investigated the contributions of this transporter to the ability of M. tuberculosis to colonize macrophages. An H37Rv proXVWZ mutant was impaired for initial survival and intracellular growth and exhibited reduced growth at elevated medium osmolarity. This defect could be complemented by restoring proXVWZ and was attributable to a failure to accumulate the compatible solute glycine betaine. We then demonstrated that ProXVWZ allows M. tuberculosis to obtain betaine from host macrophages and thereby contributes to early steps in colonizing this niche. PMID- 18390666 TI - Mobilization of the incQ plasmid R300B with a chromosomal conjugation system in Salmonella enterica serovar typhi. AB - Salmonella pathogenicity island 7 (SPI-7) in Salmonella enterica serovar Typhi appears to be related to other genomic islands. Evidence suggests that SPI-7 is susceptible to spontaneous circularization, loss, and transposition. Here, we demonstrate that a region within SPI-7 has the ability to mobilize the small incQ plasmid R300B. PMID- 18390667 TI - The Xenorhabdus nematophila nilABC genes confer the ability of Xenorhabdus spp. to colonize Steinernema carpocapsae nematodes. AB - Members of the Steinernema genus of nematodes are colonized mutualistically by members of the Xenorhabdus genus of bacteria. In nature, Steinernema carpocapsae nematodes are always found in association with Xenorhabdus nematophila bacteria. Thus, this interaction, like many microbe-host associations, appears to be species specific. X. nematophila requires the nilA, nilB, and nilC genes to colonize S. carpocapsae. In this work, we showed that of all the Xenorhabdus species examined, only X. nematophila has the nilA, nilB, and nilC genes. By exposing S. carpocapsae to other Xenorhabdus spp., we established that only X. nematophila is able to colonize S. carpocapsae; therefore, the S. carpocapsae-X. nematophila interaction is species specific. Further, we showed that introduction of the nilA, nilB, and nilC genes into other Xenorhabdus species enables them to colonize the same S. carpocapsae host tissue that is normally colonized by X. nematophila. Finally, sequence analysis supported the idea that the nil genes were horizontally acquired. Our findings indicate that a single genetic locus determines host specificity in this bacteria-animal mutualism and that host range expansion can occur through the acquisition of a small genetic element. PMID- 18390669 TI - MicroRNA-repressed mRNAs contain 40S but not 60S components. AB - MicroRNAs (miRNAs) are small noncoding RNAs that may target more than one-third of human genes, yet the mechanisms used by miRNAs to repress translation of target mRNAs are obscure. Using a recently described cell-free assay of miRNA function, we observe that miRNA-targeted mRNAs are enriched for 40S but not 60S ribosome components. Additionally, toeprinting analysis of miRNA-targeted mRNAs demonstrates that approximately 18 nt 3' relative to the initiating AUG are protected, consistent with 40S ribosome subunits positioned at the AUG codon. Our results suggest that miRNAs repress translation initiation by preventing 60S subunit joining to miRNA-targeted mRNAs. PMID- 18390668 TI - MicroRNA 29c is down-regulated in nasopharyngeal carcinomas, up-regulating mRNAs encoding extracellular matrix proteins. AB - Using highly sensitive microarray-based procedures, we identified eight microRNAs (miRNAs) showing robust differential expression between 31 laser-capture microdissected nasopharyngeal carcinomas (NPCs) and 10 normal healthy nasopharyngeal epithelial samples. In particular, miRNA mir-29c was expressed at one-fifth the levels in tumors as in normal epithelium. In NPC tumors, the lower mir-29c levels correlated with higher levels of multiple mRNAs whose 3' UTRs can bind mir-29c at target sequences conserved across many vertebrates. In cultured cells, introduction of mir-29c down-regulated these genes at the level of mRNA and inhibited expression of luciferase encoded by vectors having the 3' UTRs of these genes. Moreover, for each of several genes tested, mutating the mir-29c target sites in the 3' UTR abrogated mir-29c-induced inhibition of luciferase expression. Most of the mir-29c-targeted genes identified encode extracellular matrix proteins, including multiple collagens and laminin gamma1, that are associated with tumor cell invasiveness and metastatic potential, prominent characteristics of NPC. Thus, we identify eight miRNAs differentially expressed in NPC and demonstrate the involvement of one in regulating genes involved in metastasis. PMID- 18390670 TI - Evolutionary relationships of "Candidatus endobugula" bacterial symbionts and their Bugula bryozoan hosts. AB - Ribosomal gene sequences were obtained from bryozoans in the genus Bugula and their bacterial symbionts; analyses of host and symbiont phylogenetic trees did not support a history of strict cospeciation. Symbiont-derived compounds known to defend host larvae from predation were only detected in two out of four symbiotic Bugula species. PMID- 18390671 TI - Improvement of the glutaryl-7-aminocephalosporanic acid acylase activity of a bacterial gamma-glutamyltranspeptidase. AB - 7-Aminocephalosporanic acid (7-ACA) is an important material in the production of semisynthetic cephalosporins, which are the best-selling antibiotics worldwide. 7 ACA is produced from cephalosporin C via glutaryl-7-ACA (GL-7-ACA) by a bioconversion process using d-amino acid oxidase and cephalosporin acylase (or GL 7-ACA acylase). Previous studies demonstrated that a single amino acid substitution, D433N, provided GL-7-ACA acylase activity for gamma glutamyltranspeptidase (GGT) of Escherichia coli K-12. In this study, based on its three-dimensional structure, residues involved in substrate recognition of E. coli GGT were rationally mutagenized, and effective mutations were then combined. A novel screening method, activity staining followed by a GL-7-ACA acylase assay with whole cells, was developed, and it enabled us to obtain mutant enzymes with enhanced GL-7-ACA acylase activity. The best mutant enzyme for catalytic efficiency, with a k(cat)/K(m) value for GL-7-ACA almost 50-fold higher than that of the D433N enzyme, has three amino acid substitutions: D433N, Y444A, and G484A. We also suggest that GGT from Bacillus subtilis 168 can be another source of GL-7 ACA acylase for industrial applications. PMID- 18390672 TI - Characterization of the RND-type multidrug efflux pump MexAB-OprM of the plant pathogen Pseudomonas syringae. AB - In gram-negative bacteria, transporters belonging to the RND family are the transporters most relevant for resistance to antimicrobial compounds. In Pseudomonas aeruginosa, a clinically important pathogen, the RND-type pump MexAB OprM has been recognized as one of the major multidrug efflux systems. Here, homologues of MexAB-OprM in the plant pathogens Pseudomonas syringae pv. phaseolicola 1448A, P. syringae pv. syringae B728a, and P. syringae pv. tomato DC3000 were identified, and mexAB-oprM-deficient mutants were generated. Determination of MICs revealed that mutation of MexAB-OprM dramatically reduced the tolerance to a broad range of antimicrobials. Moreover, the ability of the mexAB-oprM-deficient mutants to multiply in planta was reduced. RNA dot blot hybridization revealed growth-dependent regulation of the mexAB-oprM operon in P. syringae; the expression of this operon was maximal in early exponential phase and decreased gradually during further growth. PMID- 18390673 TI - Factors controlling the distribution of archaeal tetraethers in terrestrial hot springs. AB - Glycerol dialkyl glycerol tetraethers (GDGTs) found in hot springs reflect the abundance and community structure of Archaea in these extreme environments. The relationships between GDGTs, archaeal communities, and physical or geochemical variables are underexamined to date and when reported often result in conflicting interpretations. Here, we examined profiles of GDGTs from pure cultures of Crenarchaeota and from terrestrial geothermal springs representing a wide distribution of locations, including Yellowstone National Park (United States), the Great Basin of Nevada and California (United States), Kamchatka (Russia), Tengchong thermal field (China), and Thailand. These samples had temperatures of 36.5 to 87 degrees C and pH values of 3.0 to 9.2. GDGT abundances also were determined for three soil samples adjacent to some of the hot springs. Principal component analysis identified four factors that accounted for most of the variance among nine individual GDGTs, temperature, and pH. Significant correlations were observed between pH and the GDGTs crenarchaeol and GDGT-4 (four cyclopentane rings, m/z 1,294); pH correlated positively with crenarchaeol and inversely with GDGT-4. Weaker correlations were observed between temperature and the four factors. Three of the four GDGTs used in the marine TEX(86) paleotemperature index (GDGT-1 to -3, but not crenarchaeol isomer) were associated with a single factor. No correlation was observed for GDGT-0 (acyclic caldarchaeol): it is effectively its own variable. The biosynthetic mechanisms and exact archaeal community structures leading to these relationships remain unknown. However, the data in general show promise for the continued development of GDGT lipid-based physiochemical proxies for archaeal evolution and for paleo ecology or paleoclimate studies. PMID- 18390674 TI - Responses of Mycobacterium tuberculosis hemoglobin promoters to in vitro and in vivo growth conditions. AB - The success of Mycobacterium tuberculosis as one of the dreaded human pathogens lies in its ability to utilize different defense mechanisms in response to the varied environmental challenges during the course of its intracellular infection, latency, and reactivation cycle. Truncated hemoglobins trHbN and trHbO are thought to play pivotal roles in the cellular metabolism of this organism during stress and hypoxia. To delineate the genetic regulation of the M. tuberculosis hemoglobins, transcriptional fusions of the promoters of the glbN and glbO genes with green fluorescent protein were constructed, and their responses were monitored in Mycobacterium smegmatis and M. tuberculosis H37Ra exposed to environmental stresses in vitro and in M. tuberculosis H37Ra after in vivo growth inside macrophages. The glbN promoter activity increased substantially during stationary phase and was nearly 3- to 3.5-fold higher than the activity of the glbO promoter, which remained more or less constant during different growth phases in M. smegmatis, as well as in M. tuberculosis H37Ra. In both mycobacterial hosts, the glbN promoter activity was induced 1.5- to 2-fold by the general nitrosative stress inducer, nitrite, as well as the NO releaser, sodium nitroprusside (SNP). The glbO promoter was more responsive to nitrite than to SNP, although the overall increase in its activity was much less than that of the glbN promoter. Additionally, the glbN promoter remained insensitive to the oxidative stress generated by H(2)O(2), but the glbO promoter activity increased nearly 1.5-fold under similar conditions, suggesting that the trHb gene promoters are regulated differently under nitrosative and oxidative stress conditions. In contrast, transition metal-induced hypoxia enhanced the activity of both the glbN and glbO promoters at all growth phases; the glbO promoter was induced approximately 2.3-fold, which was found to be the highest value for this promoter under all the conditions evaluated. Addition of iron along with nickel reversed the induction in both cases. Interestingly, a concentration-dependent decrease in the activity of both trHb gene promoters was observed when the levels of iron in the growth media were depleted by addition of an iron chelator. These results suggested that an iron/heme-containing oxygen sensor is involved in the modulation of the trHb gene promoter activities directly or indirectly in conjunction with other cellular factors. The modes of promoter regulation under different physiological conditions were found to be similar for the trHbs in both M. smegmatis and M. tuberculosis H37Ra, indicating that the promoters might be regulated by components that are common to the two systems. Confocal microscopy of THP-1 macrophages infected with M. tuberculosis carrying the trHb gene promoter fusions showed that there was a significant level of promoter activity during intracellular growth in macrophages. Time course evaluation of the promoter activity after various times up to 48 h by fluorescence-activated cell sorting analysis of the intracellular M. tuberculosis cells indicated that the glbN promoter was active at all time points assessed, whereas the activity of the glbO promoter remained at a steady-state level up to 24 h postinfection and increased approximately 2-fold after 48 h of infection. Thus, the overall regulation pattern of the M. tuberculosis trHb gene promoters correlates not only with the stresses that the tubercle bacillus is likely to encounter once it is in the macrophage environment but also with our current knowledge of their functions. The in vivo studies that demonstrated for the first time expression of trHbs during macrophage infection of M. tuberculosis strongly indicate that the hemoglobins are required, and thus important, during the intracellular phase of the bacterial cycle. The present study of transcriptional regulation of M. tuberculosis hemoglobins in vitro under various stress conditions and in vivo after macrophage infection supports the hypothesis that biosynthesis of both trHbs (trHbN and trHbO) in the native host is regulated via the environmental signals that the tubercle bacillus receives during macrophage infection and growth in its human host. PMID- 18390675 TI - Transcriptional analysis of L-methionine catabolism in the cheese-ripening yeast Yarrowia lipolytica in relation to volatile sulfur compound biosynthesis. AB - Yarrowia lipolytica is one of the yeasts most frequently isolated from the surface of ripened cheeses. In previous work, it has been shown that this yeast is able to convert L-methionine into various volatile sulfur compounds (VSCs) that may contribute to the typical flavors of several cheeses. In the present study, we show that Y. lipolytica does not assimilate lactate in the presence of L-methionine in a cheeselike medium. Nineteen presumptive genes associated with L methionine catabolism or pyruvate metabolism in Y. lipolytica were transcriptionally studied in relation to L-methionine degradation. The expression levels of the YlARO8 (YALI0E20977g), YlBAT1 (YALI0D01265g), and YlBAT2 (YALI0F19910g) genes (confirmed by real-time PCR experiments) were found to be strongly up-regulated by L-methionine, and a greater variety and larger amounts of VSCs, such as methanethiol and its autooxidation products (dimethyl disulfide and dimethyl trisulfide), were released in the medium when Y. lipolytica was grown in the presence of a high concentration of L-methionine. In contrast, other genes related to pyruvate metabolism were found to be down-regulated in the presence of L-methionine; two exceptions were the YlPDB1 (YALI0E27005g) and YlPDC6 (YALI0D06930g) genes, which encode a pyruvate dehydrogenase and a pyruvate decarboxylase, respectively. Both transcriptional and biochemical results corroborate the view that transamination is the first step of the enzymatic conversion of L-methionine to VSCs in Y. lipolytica and that the YlARO8, YlBAT1, and YlBAT2 genes could play a key role in this process. PMID- 18390676 TI - Complete genome sequence of Nitrosospira multiformis, an ammonia-oxidizing bacterium from the soil environment. AB - The complete genome of the ammonia-oxidizing bacterium Nitrosospira multiformis (ATCC 25196(T)) consists of a circular chromosome and three small plasmids totaling 3,234,309 bp and encoding 2,827 putative proteins. Of the 2,827 putative proteins, 2,026 proteins have predicted functions and 801 are without conserved functional domains, yet 747 of these have similarity to other predicted proteins in databases. Gene homologs from Nitrosomonas europaea and Nitrosomonas eutropha were the best match for 42% of the predicted genes in N. multiformis. The N. multiformis genome contains three nearly identical copies of amo and hao gene clusters as large repeats. The features of N. multiformis that distinguish it from N. europaea include the presence of gene clusters encoding urease and hydrogenase, a ribulose-bisphosphate carboxylase/oxygenase-encoding operon of distinctive structure and phylogeny, and a relatively small complement of genes related to Fe acquisition. Systems for synthesis of a pyoverdine-like siderophore and for acyl-homoserine lactone were unique to N. multiformis among the sequenced genomes of ammonia-oxidizing bacteria. Gene clusters encoding proteins associated with outer membrane and cell envelope functions, including transporters, porins, exopolysaccharide synthesis, capsule formation, and protein sorting/export, were abundant. Numerous sensory transduction and response regulator gene systems directed toward sensing of the extracellular environment are described. Gene clusters for glycogen, polyphosphate, and cyanophycin storage and utilization were identified, providing mechanisms for meeting energy requirements under substrate-limited conditions. The genome of N. multiformis encodes the core pathways for chemolithoautotrophy along with adaptations for surface growth and survival in soil environments. PMID- 18390677 TI - Detection and quantification of group C rotaviruses in communal sewage. AB - Group C rotaviruses have been recognized as a cause of acute gastroenteritis in humans, cattle, and swine, although the true epidemiologic and clinical importance of this virus in these hosts has not yet been fully established. A real-time PCR assay based on a broadly reactive primer pair was developed and used to quantitatively determine the viral load of group C rotaviruses in environmental samples. A total of 35 raw and 35 treated sewage samples collected at the same sampling time in four Hungarian sewage treatment plants during a survey in 2005 were tested for the presence of group C rotaviruses. The overall detection rates were 91% (32 of 35) for the influent and 57% (20 of 35) for the effluent samples. Molecular characterization of the amplified partial VP6 gene revealed the cocirculation of human and animal (i.e., bovine and porcine) strains that were easily distinguishable by melting curve analysis. Human strains yielded relatively high viral loads (mean, 1.2 x 10(7); median, 6.9 x 10(5) genome equivalents per liter influent sewage) and appeared to display seasonal activity over the study period, whereas animal strains appeared to circulate throughout the year at much lower average titers (bovine strains mean, 9.9 x 10(4); median, 3.0 x 10(4); porcine strains mean, 3.9 x 10(4); median, 3.1 x 10(4) genome equivalents per liter influent sewage). Our findings suggest that monitoring of communal sewage may provide a good surrogate for investigating the epidemiology and ecology of group C rotaviruses in humans and animals. PMID- 18390678 TI - Characterization and genomic analysis of phage asccphi28, a phage of the family Podoviridae infecting Lactococcus lactis. AB - Bacteriophage asccphi28 infects dairy fermentation strains of Lactococcus lactis. This report describes characterization of asccphi28 and its full genome sequence. Phage asccphi28 has a prolate head, whiskers, and a short tail (C2 morphotype). This morphology and DNA hybridization to L. lactis phage P369 DNA showed that asccphi28 belongs to the P034 phage species, a group rarely encountered in the dairy industry. The burst size of asccphi28 was found to be 121 +/- 18 PFU per infected bacterial cell after a latent period of 44 min. The linear genome (18,762 bp) contains 28 possible open reading frames (ORFs) comprising 90% of the total genome. The ORFs are arranged bidirectionally in recognizable functional modules. The genome contains 577 bp inverted terminal repeats (ITRs) and putatively eight promoters and four terminators. The presence of ITRs, a phage encoded DNA polymerase, and a terminal protein that binds to the DNA, along with BLAST and morphology data, show that asccphi28 more closely resembles streptococcal phage Cp-1 and the phi29-like phages that infect Bacillus subtilis than it resembles common lactococcal phages. The sequence of this phage is the first published sequence of a P034 species phage genome. PMID- 18390679 TI - High-yield production of secreted active proteins by the Pseudomonas aeruginosa type III secretion system. AB - The Escherichia coli system is the system of choice for recombinant protein production because it is possible to obtain a high protein yield in inexpensive media. The accumulation of protein in an insoluble form in inclusion bodies remains a major disadvantage. Use of the Pseudomonas aeruginosa type III secretion system can avoid this problem, allowing the production of soluble secreted proteins. PMID- 18390680 TI - Inactivation of Bacillus anthracis spores by a combination of biocides and heating under high-temperature short-time pasteurization conditions. AB - The milk supply is considered a primary route for a bioterrorism attack with Bacillus anthracis spores because typical high-temperature short-time (HTST) pasteurization conditions cannot inactivate spores. In the event of intentional contamination, an effective method to inactivate the spores in milk under HTST processing conditions is needed. This study was undertaken to identify combinations and concentrations of biocides that can inactivate B. anthracis spores at temperatures in the HTST range in less than 1 min. Hydrogen peroxide (HP), sodium hypochlorite (SH), and peroxyacetic acid (PA) were evaluated for their efficacy in inactivating spores of strains 7702, ANR-1, and 9131 in milk at 72, 80, and 85 degrees C using a sealed capillary tube technique. Strains ANR-1 and 9131 were more resistant to all of the biocide treatments than strain 7702. Addition of 1,260 ppm SH to milk reduced the number of viable spores of each strain by 6 log CFU/ml in less than 90 and 60 s at 72 and 80 degrees C, respectively. After neutralization, 1,260 ppm SH reduced the time necessary to inactivate 6 log CFU/ml (TTI6-log) at 80 degrees C to less than 20 s. Treatment of milk with 7,000 ppm HP resulted in a similar level of inactivation in 60 s. Combined treatment with 1,260 ppm SH and 1,800 ppm HP inactivated spores of all strains in less than 20 s at 80 degrees C. Mixing 15 ppm PA with milk containing 1,260 ppm SH resulted in TTI6-log of 25 and 12 s at 72 and 80 degrees C, respectively. TTI6-log of less than 20 s were also achieved at 80 degrees C by using two combinations of biocides: 250 ppm SH, 700 ppm HP, and 150 ppm PA; and 420 ppm SH (pH 7), 1,100 ppm HP, and 15 ppm PA. These results indicated that different combinations of biocides could consistently result in 6-log reductions in the number of B. anthracis spores in less than 1 min at temperatures in the HTST range. This information could be useful for developing more effective thermal treatment strategies which could be used in HTST milk plants to process contaminated milk for disposal and decontamination, as well as for potential protective measures. PMID- 18390681 TI - Characterization of replication and conjugation of Streptomyces circular plasmids pFP1 and pFP11 and their ability to propagate in linear mode with artificially attached telomeres. AB - Many Streptomyces species harbor circular plasmids (8 to 31 kb) as well as linear plasmids (12 to 1,700 kb). We report the characterization of two newly detected circular plasmids, pFP11 (35,139 bp) and pFP1 (39,360 bp). As on linear plasmids, their replication loci comprise repA genes and adjacent iterons, to which RepA proteins bind specifically in vitro. Plasmids containing the minimal iterons plus the repA locus of pFP11 were inherited extremely unstably; par and additional loci were required for stable inheritance. Surprisingly, plasmids containing replication loci from pFP11 or Streptomyces circular plasmid SCP2 but not from pFP1, SLP1, or pIJ101 propagated in a stable linear mode when the telomeres of a linear plasmid were attached. These results indicate bidirectional replication for pFP11 and SCP2. Both pFP11 and pFP1 contain, for plasmid transfer, a major functional traB gene (encoding a DNA translocase typical for Streptomyces plasmids) as well as, surprisingly, a putative traA gene (encoding a DNA nickase, characteristic of single-stranded DNA transfer of gram-negative plasmids), but this did not appear to be functional, at least in isolation. PMID- 18390682 TI - Bayesian-integrated microbial forensics. AB - In the aftermath of the 2001 anthrax letters, researchers have been exploring ways to predict the production environment of unknown-source microorganisms. Culture medium, presence of agar, culturing temperature, and drying method are just some of the broad spectrum of characteristics an investigator might like to infer. The effects of many of these factors on microorganisms are not well understood, but the complex way in which microbes interact with their environments suggests that numerous analytical techniques measuring different properties will eventually be needed for complete characterization. In this work, we present a Bayesian statistical framework for integrating disparate analytical measurements. We illustrate its application to the problem of characterizing the culture medium of Bacillus spores using three different mass spectral techniques. The results of our study suggest that integrating data in this way significantly improves the accuracy and robustness of the analyses. PMID- 18390683 TI - Dynamics of Legionella spp. and bacterial populations during the proliferation of L. pneumophila in a cooling tower facility. AB - The dynamics of Legionella spp. and of dominant bacteria were investigated in water from a cooling tower plant over a 9-month period which included several weeks when Legionella pneumophila proliferated. The structural diversity of both the bacteria and the Legionella spp. was monitored by a fingerprint technique, single-strand conformation polymorphism, and Legionella spp. and L. pneumophila were quantified by real-time quantitative PCR. The structure of the bacterial community did not change over time, but it was perturbed periodically by chemical treatment or biofilm detachment. In contrast, the structure of the Legionella sp. population changed in different periods, its dynamics at times showing stability but also a rapid major shift during the proliferation of L. pneumophila in July. The dynamics of the Legionella spp. and of dominant bacteria were not correlated. In particular, no change in the bacterial community structure was observed during the proliferation of L. pneumophila. Legionella spp. present in the cooling tower system were identified by cloning and sequencing of 16S rRNA genes. A high diversity of Legionella spp. was observed before proliferation, including L. lytica, L. fallonii, and other Legionella-like amoebal pathogen types, along with as-yet-undescribed species. During the proliferation of L. pneumophila, Legionella sp. diversity decreased significantly, L. fallonii and L. pneumophila being the main species recovered. PMID- 18390684 TI - Comparison of Campylobacter populations in wild geese with those in starlings and free-range poultry on the same farm. AB - Wild geese are a potential source of Campylobacter infection for humans and farm animals and have been implicated in at least two large waterborne disease outbreaks. There have been few investigations into the population biology of Campylobacter in geese, carriage rates are reported to vary (0 to 100%), and no genetic characterization of isolates has been performed. Fecal samples collected from wild geese in Oxfordshire, United Kingdom, were culture positive for C. jejuni (50.2%) and C. coli (0.3%). The C. jejuni (n = 166) isolates were characterized by using multilocus sequence typing and were compared with isolates collected from free-range broiler chickens and wild starlings sampled at the same location. A total of 38 STs, six clonal complexes, and 23 flaA SVR nucleotide STs were identified. The ST-21 and ST-45 complexes (5.4% of isolates) were the only complexes to be identified among isolates from the geese and the other bird species sampled in the same location. These clonal complexes were also identified among human disease isolates collected in the same health care region. The results indicate that large numbers of wild geese carry Campylobacter; however, there was limited mixing of Campylobacter populations among the different sources examined, and the host source could be predicted with high probability from the allelic profile of a C. jejuni isolate. In conclusion, genotypes of C. jejuni isolated from geese are highly host specific, and a comparison with isolates from Oxfordshire cases of human disease revealed that while geese cannot be excluded as a source of infection for humans and farm animals, their contribution is likely to be minor. PMID- 18390685 TI - Characterization of endogenous plasmids from Lactobacillus salivarius UCC118. AB - The genome of Lactobacillus salivarius UCC118 comprises a 1.83-Mb chromosome, a 242-kb megaplasmid (pMP118), and two smaller plasmids of 20 kb (pSF118-20) and 44 kb (pSF118-44). Annotation and bioinformatic analyses suggest that both of the smaller plasmids replicate by a theta replication mechanism. Furthermore, it appears that they are transmissible, although neither possesses a complete set of conjugation genes. Plasmid pSF118-20 encodes a toxin-antitoxin system composed of pemI and pemK homologs, and this plasmid could be cured when PemI was produced in trans. The minimal replicon of pSF118-20 was determined by deletion analysis. Shuttle vector derivatives of pSF118-20 were generated that included the replication region (pLS203) and the replication region plus mobilization genes (pLS208). The plasmid pLS203 was stably maintained without selection in Lactobacillus plantarum, Lactobacillus fermentum, and the pSF118-20-cured derivative strain of L. salivarius UCC118 (strain LS201). Cloning in pLS203 of genes encoding luciferase and green fluorescent protein, and expression from a constitutive L. salivarius promoter, demonstrated the utility of this vector for the expression of heterologous genes in Lactobacillus. This study thus expands the knowledge base and vector repertoire of probiotic lactobacilli. PMID- 18390686 TI - Effect of germ cell depletion on levels of specific mRNA transcripts in mouse Sertoli cells and Leydig cells. AB - It has been shown that testicular germ cell development is critically dependent upon somatic cell activity but, conversely, the extent to which germ cells normally regulate somatic cell function is less clear. This study was designed, therefore, to examine the effect of germ cell depletion on Sertoli cell and Leydig cell transcript levels. Mice were treated with busulphan to deplete the germ cell population and levels of mRNA transcripts encoding 26 Sertoli cell specific proteins and 6 Leydig cell proteins were measured by real-time PCR up to 50 days after treatment. Spermatogonia were lost from the testis between 5 and 10 days after treatment, while spermatocytes were depleted after 10 days and spermatids after 20 days. By 30 days after treatment, most tubules were devoid of germ cells. Circulating FSH and intratesticular testosterone were not significantly affected by treatment. Of the 26 Sertoli cell markers tested, 13 showed no change in transcript levels after busulphan treatment, 2 showed decreased levels, 9 showed increased levels and 2 showed a biphasic response. In 60% of cases, changes in transcript levels occurred after the loss of the spermatids. Levels of mRNA transcripts encoding Leydig cell-specific products related to steroidogenesis were unaffected by treatment. Results indicate (1) that germ cells play a major and widespread role in the regulation of Sertoli cell activity, (2) most changes in transcript levels are associated with the loss of spermatids and (3) Leydig cell steroidogenesis is largely unaffected by germ cell ablation. PMID- 18390687 TI - Two pathways for prostaglandin F2 alpha synthesis by the primate periovulatory follicle. AB - Prostaglandin E2 (PGE2) has been identified as a PG necessary for ovulation, but the ovulatory gonadotropin surge also increases PGF2 alpha levels in primate periovulatory follicles. To better understand the role of PGF2 alpha in ovulation, pathways utilized for PGF2 alpha synthesis by the primate follicle were examined. Monkeys were treated with gonadotropins to stimulate multiple follicular development; follicular aspirates and whole ovaries were removed before and at specific times after administration of an ovulatory dose of hCG to span the 40 h periovulatory interval. Human granulosa cells were also obtained (typically 34-36 h after hCG) from in vitro fertilization patients. PGF2 alpha can be synthesized from PGH2 via the aldo-keto reductase (AKR) 1C3. AKR1C3 mRNA and protein levels in monkey granulosa cells were low before hCG and peaked 24-36 h after hCG administration. Human granulosa cells converted PGD2 into 11 beta PGF2 alpha, confirming that these cells possess AKR1C3 activity. PGF2 alpha can also be synthesized from PGE2 via the enzymes AKR1C1 and AKR1C2. Monkey granulosa cell levels of AKR1C1/AKR1C2 mRNA was low 0-12 h, peaked at 24 h, and returned to low levels by 36 h after hCG administration. Human granulosa cell conversion of [(3)H]PGE2 into [(3)H]PGF2 alpha was reduced by an AKR1C2-selective inhibitor, supporting the concept that granulosa cells preferentially express AKR1C2 over AKR1C1. In summary, the ovulatory gonadotropin surge increases granulosa cell expression of AKR1C1/AKR1C2 and AKR1C3. Both of these enzyme activities are present in periovulatory granulosa cells. These data support the concept that follicular PGF2 alpha can be synthesized via two pathways during the periovulatory interval. PMID- 18390688 TI - Expression of the pluripotency marker UTF1 is restricted to a subpopulation of early A spermatogonia in rat testis. AB - The population of early A spermatogonia includes stem cells that possess spermatogonial stem cell properties. Recent reports suggest that these cells have the ability to regain pluripotent properties. Here, we show that expression of the pluripotency marker undifferentiated embryonic cell transcription factor 1 (UTF1) is restricted to distinct germ cells within the testis. In embryonic and neonatal testes, all gonocytes were found to strongly express UTF1. During further testicular development, expression of UTF1 was restricted to a subset of A spermatogonia and with the increase in age the number of cells expressing UTF1 decreased even more. Ultimately, in the adult rat testis, only a small subset of the A spermatogonia expressed UTF1. Remarkably, even in testes of vitamin A deficient rats, in which the early A spermatogonia (A(s), A(pr), and A(al)) are the only type of spermatogonia, only a subset of the spermatogonia expressed UTF1. In the adult rat testis, expression of UTF1 is restricted to a subpopulation of the ZBTB16 (PLZF)-positive early A spermatogonia. Furthermore, the observed distribution pattern of UTF1-expressing cells over the different stages of the cycle of the seminiferous epithelium suggests that the expression of UTF1 is restricted to those A(s), A(pr), and short chains of A(al) spermatogonia that are in the undifferentiated state and therefore maintain the ability to differentiate into A1 spermatogonia in the next round of the epithelial cycle or possibly even in other directions when they are taken out of their testicular niche. PMID- 18390690 TI - Regulation of endothelial proliferation by the renin-angiotensin system in human umbilical vein endothelial cells. AB - This study was performed in order to evaluate the role of angiotensin II in physiological angiogenesis. Human umbilical vein endothelial cells (HUVEC) were stained for angiotensin II type 1 receptor (AGTR1) immunocytochemically and for gene expression of renin-angiotensin system (RAS) components. The regulation of the angiogenesis-associated genes vascular endothelial growth factor (VEGF) and angiopoietins (ANGPT1 and ANGPT2) were studied using quantitative RT-PCR. Furthermore, we examined the effect of angiotensin II on the proliferation of HUVEC using Ki-67 as well as BrdU immunocytochemistry and investigated whether the administration of the AGTR1 blocker candesartan or the VEGF antagonist FLT1 Fc could suppress the observed angiotensin II-dependent proangiogenic effect. AGTR1 was expressed in HUVEC and the administration of angiotensin II significantly increased the gene expression of VEGF and decreased the gene expression of ANGPT1. Since the expression of ANGPT2 was not affected significantly the ratio of ANGPT1/ANGPT2 was decreased. In addition, a significantly increased endothelial cell proliferation was observed after stimulation with angiotensin II, which was suppressed by the simultaneous administration of candesartan or the VEGF antagonist FLT1-Fc. These results indicate the potential capacity of angiotensin II in influencing angiogenesis by the regulation of angiogenesis-associated genes via AGTR1. Since VEGF blockade opposed the effect of angiotensin II on cell proliferation, it is hypothesised that VEGF mediates the angiotensin II-dependent effect in concert with the changes in angiopoietin expression. This is the first report of the RAS on the regulation of angiogenesis-associated genes in physiology. PMID- 18390689 TI - Estrous cycle, pregnancy, and parity enhance performance of rats in object recognition or object placement tasks. AB - Ovarian hormone elevations are associated with enhanced learning/memory. During behavioral estrus or pregnancy, progestins, such as progesterone (P(4)) and its metabolite 5 alpha-pregnan-3 alpha-ol-20-one (3 alpha,5 alpha-THP), are elevated due, in part, to corpora luteal and placental secretion. During 'pseudopregnancy', the induction of corpora luteal functioning results in a hormonal milieu analogous to pregnancy, which ceases after about 12 days, due to the lack of placental formation. Multiparity is also associated with enhanced learning/memory, perhaps due to prior steroid exposure during pregnancy. Given evidence that progestins and/or parity may influence cognition, we investigated how natural alterations in the progestin milieu influence cognitive performance. In Experiment 1, virgin rats (nulliparous) or rats with two prior pregnancies (multiparous) were assessed on the object placement and recognition tasks, when in high-estrogen/P(4) (behavioral estrus) or low-estrogen/P(4) (diestrus) phases of the estrous cycle. In Experiment 2, primiparous or multiparous rats were tested in the object placement and recognition tasks when not pregnant, pseudopregnant, or pregnant (between gestational days (GDs) 6 and 12). In Experiment 3, pregnant primiparous or multiparous rats were assessed daily in the object placement or recognition tasks. Females in natural states associated with higher endogenous progestins (behavioral estrus, pregnancy, multiparity) outperformed rats in low progestin states (diestrus, non-pregnancy, nulliparity) on the object placement and recognition tasks. In earlier pregnancy, multiparous, compared with primiparous, rats had a lower corticosterone, but higher estrogen levels, concomitant with better object placement performance. From GD 13 until post partum, primiparous rats had higher 3 alpha,5 alpha-THP levels and improved object placement performance compared with multiparous rats. PMID- 18390691 TI - A single, mild, transient scrotal heat stress causes DNA damage, subfertility and impairs formation of blastocysts in mice. AB - Infertility represents a major clinical problem and 50% of cases are attributable to the male partner. Testicular function is temperature dependent, and in both man and mouse the position of the testes in the scrotum ensures that they are kept at between 2 and 8 degrees C below core body temperature. We used a mouse model to investigate the impact of a single, transient, mild, scrotal heat stress (38, 40 or 42 degrees C for 30 min) on testicular function, sperm DNA integrity and embryo survival. We detected temperature-dependent changes in testicular architecture, number of apoptotic cells and a significant reduction in testis weight 7 and 14 days after heat stress at 42 degrees C. We report for the first time that DNA strand breaks (gamma-H2AX-positive foci) were present in spermatocytes recovered from testes subjected to 40 or 42 degrees C. Fertility of heat-stressed males was tested 23-28 d after treatment (sperm at this time would have been spermatocytes at time of heating). Paternal heat stress at 42 degrees C resulted in reduced pregnancy rate, placental weight and litter size; pregnancies from the 40 degrees C group had increased resorptions at e14.5. Abnormalities in embryonic development were detected at e3.5 and in vitro fertilisation with sperm recovered 16 h or 23 d after scrotal stress at 42 degrees C revealed a block in development between the 4-cell and blastocyst stages. This study has provided evidence of temperature-dependent effects on germ cell DNA integrity and highlighted the importance of an intact paternal genome for normal embryo development. PMID- 18390692 TI - Presence, processing, and localization of mouse ADAM15 during sperm maturation and the role of its disintegrin domain during sperm-egg binding. AB - Successful fertilization requires gametes to complete several stages, beginning with maturation and transport along the male and female reproductive tracts and ending with the interaction between the sperm and the egg. This last step involves sperm-egg adhesion and membrane fusion. ADAMs (disintegrin and metalloprotease domain proteins) are a family of membrane-anchored glycoproteins that are thought to play diverse roles in cell-cell adhesion through their interaction with integrins. This study analyzes the presence, location, processing, and possible role of ADAM15 in mouse sperm. The presence of ADAM15 in mouse spermatozoa was detected by Western blotting, which revealed that ADAM15 is post-translationally processed, during epididymal sperm maturation and the acrosome reaction. The 35 kDa antigen present in the acrosome-reacted sperm is the last proteolytic product of the 110/75 kDa ADAM15 found in non-capacitated sperm. This 35 kDa protein contains the disintegrin domain. By indirect immunofluorescence, ADAM15 was identified in the acrosomal region and along the flagellum of mouse spermatozoa. In acrosome-reacted sperm, ADAM15 was lost from the acrosomal region, but remained diffusely distributed throughout the head and flagellum. Furthermore, the ADAM15 disintegrin domain (RPPTDDCDLPEF) partially inhibited fusion and almost completely inhibited sperm-oolemma adhesion. In conclusion, our data indicate that ADAM15 is present in the testis and in spermatozoa from the caput, corpus, and cauda epididymis, as well as in non capacitated and acrosome-reacted gametes. Results also indicate that ADAM15 is processed during epididymal maturation and acrosome reaction and that it may play a role during sperm-egg binding. PMID- 18390693 TI - Xenografting of sheep testis tissue and isolated cells as a model for preservation of genetic material from endangered ungulates. AB - Recovery of germ cells could be an option for preservation of the genetic pool of endangered animals. In immature males, xenografting of testis tissue provides the opportunity to recover sperm from these animals. In adult animals, xenografting has been less successful, but de novo morphogenesis of functional testis tissue from dissociated testis cells could be an alternative. To assess the potential use of these techniques in endangered bovid species, the domestic sheep was used as a model. Testes from 2-week-old lambs were grafted as tissue fragments or cell suspensions into nude mice. Grafts were recovered at 4, 8, 12 and 16 weeks post grafting. For isolated cells, two additional time points at 35 and 40 weeks after grafting were added. In addition, to analyse the possible effect of social stress among mice within a group on the development of the grafts, testis tissue grafts were recovered 13 weeks post grafting from mice housed individually and in groups. Complete spermatogenesis occurred in sheep testis xenografts at 12 weeks, similar to the situation in situ. Isolated sheep testis cells were able to reorganize and form functional testicular tissue de novo. Housing mice individually or in groups did not have any effect on the development of xenografts. Xenografting of testis tissue might be useful to obtain sperm from immature endangered ungulates that die prematurely. Testis tissue de novo morphogenesis from isolated cells could open interesting options to recover germ cells from mature males with impaired spermatogenesis. PMID- 18390694 TI - Large-scale analysis of gene clustering in bacteria. AB - An important strategy to study operons and their evolution is to investigate clustering of related genes across multiple bacterial genomes. Although existing algorithms are available that can identify gene clusters across two or more genomes, very few algorithms are efficient enough to study gene clusters across hundreds of genomes. We observe that a querying strategy can be used to analyze gene clusters across a large number of genomes and develop an efficient algorithm to identify all related clusters on a genome from a given query cluster. We use this algorithm to study gene clustering in 400 bacterial genomes by starting from a well-characterized list of operons in Escherichia coli K12 and perform comparative analysis of operon occurrences, gene orientations, and rearrangements both within and across clusters. We show that important biological insights can be obtained by comparing results across these categories. A software program implementing the algorithm (GCQuery) and supplementary data containing detailed results are available at http://faculty.cs.tamu.edu/shsze/gcquery. PMID- 18390695 TI - IL-17 and therapeutic kynurenines in pathogenic inflammation to fungi. AB - Largely viewed as proinflammatory, innate responses combine with adaptive immunity to generate the most effective form of antifungal resistance, and T cells exercise feedback control over diverse effects of inflammation on infection. Some degree of inflammation is required for protection, particularly in mucosal tissues, during the transitional response occurring between the rapid innate and slower adaptive response. However, progressive inflammation worsens disease and ultimately prevents pathogen eradication. IDO, tryptophan catabolites ("kynurenines"), and regulatory T cells help to tame overzealous and exaggerated inflammatory responses. In this context, IL-23 and the Th17 pathway, which down regulate tryptophan catabolism, may instead favor pathology and serve to accommodate the seemingly paradoxical association of chronic inflammation with fungal persistence. Recent data support a view in which IL-23/IL-17 antagonistic strategies, including the administration of synthetic kynurenines, could represent a new means of harnessing progressive or potentially harmful inflammation. PMID- 18390696 TI - Cutting edge: Broad expression of the FoxP3 locus in epithelial cells: a caution against early interpretation of fatal inflammatory diseases following in vivo depletion of FoxP3-expressing cells. AB - Dogma that the regulatory T cell (Treg) prevents catastrophic autoimmunity throughout the lifespan relies on the assumption that the FoxP3 locus is transcribed exclusively in Treg. To test the assumption, we used the Rag2(-/-) and the Rag2(-/-) mice with the Scurfy (sf) mutation (FoxP3(sf/Y) or FoxP3(sf/sf)) to evaluate FoxP3 expression outside of the lymphoid system. Immunohistochemistry and real-time PCR revealed FoxP3 expression in breast epithelial cells, lung respiratory epithelial cells, and prostate epithelial cells, although not in liver, heart, and intestine. The specificity of the assays was confirmed, as the signals were ablated by the Scurfy mutation of the FoxP3 gene. Using mice with a green fluorescence protein open reading frame knocked into the 3' untranslated region of the FoxP3 locus, we showed that the locus is transcribed broadly in epithelial cells of multiple organs. These results refute an essential underlying assumption of the dogma and question the specificity of FoxP3-based Treg depletion. PMID- 18390697 TI - Cutting edge: NKT cells constitutively express IL-23 receptor and RORgammat and rapidly produce IL-17 upon receptor ligation in an IL-6-independent fashion. AB - Th17 cells require IL-6 and TGFbeta for lineage commitment and IL-23 for maintenance. Unexpectedly, naive IL-6(-/-) splenocytes stimulated with anti-CD3 and IL-23 produced normal amounts of IL-17 during the first 24 h of culture. These rapid IL-6-independent IL-17 producers were identified as predominantly DX5(+) TCRbeta(+) NKT cells, and a comparable response could be found using the invariant NKT-specific ligand alpha-galactosylceramide. Human NKT cells also produced IL-17. NKT cells constitutively expressed IL-23R and RORgammat. Ligation of either TCR or IL-23R triggered IL-17 production and both together had a synergistic effect, suggesting independent but convergent pathways. IL-17 production was not restricted to a particular subset of NKT cells but they were NK1.1 negative. Importantly, in vivo administration of alpha-galactosylceramide triggered a rapid IL-17 response in the spleen. These data suggest an important biological role for innate IL-17 production by NKT cells that is rapid and precedes the adaptive IL-17 response. PMID- 18390698 TI - Cutting edge: Immunosuppressant as adjuvant for tolerogenic immunization. AB - Vaccination for autoimmune and alloimmune diseases has long been an attractive idea. Yet, there is no suitable adjuvant to forcefully steer the immune response toward tolerance. In this study we show that dexamethasone, a potent glucocorticoid immunosuppressant, can function as a tolerogenic adjuvant when applied together with peptide immunogen. BALB/c mice with pre-established delayed type hypersensitivity to hen OVA were immunized with an OVA-derived, MHC II restricted peptide (OVA(323-339)) in the presence of dexamethasone. The treatment caused long-term desensitization in treated animals to hen OVA via a dexamethasone-dependent tolerogenic mechanism that blocks maturation of dendritic cells and expands OVA(323-339)-specific CD4(+)CD25(+)Foxp3(+) regulatory T cells in vivo. Similar treatment of NOD mice using dexamethasone and an insulin derived, MHC II-restricted peptide (B:9-23) prevented predisposed spontaneous diabetes. Remarkably, in both models, dexamethasone-augmented immunization induced long-term persistent, Ag-specific regulatory T cells responsive to recall Ags. These results reveal for the first time the potential usefulness of immunosuppressants as tolerogenic adjuvants. PMID- 18390699 TI - Development of either split tolerance or robust tolerance along with humoral tolerance to donor and third-party alloantigens in nonmyeloablative mixed chimeras. AB - Hematopoietic chimerism is considered to generate robust allogeneic tolerance; however, tissue rejection by chimeras can occur. This "split tolerance" can result from immunity toward tissue-specific Ags not expressed by hematopoietic cells. Known to occur in chimeric recipients of skin grafts, it has not often been reported for other donor tissues. Because chimerism is viewed as a potential approach to induce islet transplantation tolerance, we generated mixed bone marrow chimerism in the tolerance-resistant NOD mouse and tested for split tolerance. An unusual multilevel split tolerance developed in NOD chimeras, but not chimeric B6 controls. NOD chimeras demonstrated persistent T cell chimerism but rejected other donor hematopoietic cells, including B cells. NOD chimeras also showed partial donor alloreactivity. Furthermore, NOD chimeras were split tolerant to donor skin transplants and even donor islet transplants, unlike control B6 chimeras. Surprisingly, islet rejection was not a result of autoimmunity, since NOD chimeras did not reject syngeneic islets. Split tolerance was linked to non-MHC genes of the NOD genetic background and was manifested recessively in F(1) studies. Also, NOD chimeras but not B6 chimeras could generate serum alloantibodies, although at greatly reduced levels compared with nonchimeric controls. Surprisingly, the alloantibody response was sufficiently cross-reactive that chimerism-induced humoral tolerance extended to third-party cells. These data identify split tolerance, generated by a tolerance-resistant genetic background, as an important new limitation to the chimerism approach. In contrast, the possibility of humoral tolerance to multiple donors is potentially beneficial. PMID- 18390701 TI - Differential regulation of human IL-7 receptor alpha expression by IL-7 and TCR signaling. AB - IL-7Ralpha is essential for the development and homeostatic maintenance of mature T cells. Studies in humans and mice have shown that IL-7Ralpha expression is reduced by its cognate cytokine, IL-7, and Ag, suggesting that active regulation of IL-7 responsiveness is necessary to balance T cell numbers. We show that IL-7- or TCR/CD28-mediated signaling induced a rapid down-regulation of IL-7Ralpha expression on naive T cells on the mRNA and protein level, with a mild (10-fold) or strong (50-fold) gene suppression, respectively. In both situations, the down regulation of IL-7Ralpha was blocked by cyclohexamide and actinomycin D, indicating the involvement of an active mechanism dependent on new transcription and protein synthesis. Upon IL-7 withdrawal, IL-7Ralpha mRNA and surface protein reappeared in a transcription-dependent manner within 7 h. Yet, IL-7Ralpha was hardly re-expressed during the same period after TCR/CD28-activation. Likewise, T cells that were activated through CMV in vivo did not re-express IL-7Ralpha after in vitro culture. Functionally, IL-7-induced down-regulation of IL-7Ralpha did not hinder the responsiveness of naive T cells to IL-7. Conversely, down regulation of IL-7Ralpha on TCR/CD28-activated cells limited IL-7 responsiveness. Strikingly, ectopic expression of IL-7Ralpha cells on TCR/CD28-activated cells conferred a selective advantage in the response to IL-7. In conclusion, our data show that IL-7- and TCR/CD28-mediated signaling differentially regulate IL 7Ralpha expression on human T cells with a transient and chronic effect, respectively. The stringent and active regulation of IL-7Ralpha may constitute a homeostatic mechanism to curtail unwarranted T cell expansion. PMID- 18390700 TI - Low-dose tolerance is mediated by the microfold cell ligand, reovirus protein sigma1. AB - Mucosal tolerance induction generally requires multiple or large Ag doses. Because microfold (M) cells have been implicated as being important for mucosal tolerance induction and because reovirus attachment protein sigma1 (psigma1) is capable of binding M cells, we postulated that targeting a model Ag to M cells via psigma1 could induce a state of unresponsiveness. Accordingly, a genetic fusion between OVA and the M cell ligand, reovirus psigma1, termed OVA-psigma1, was developed to enhance tolerogen uptake. When applied nasally, not parenterally, as little as a single dose of OVA-psigma1 failed to induce OVA specific Abs even in the presence of adjuvant. Moreover, the mice remained unresponsive to peripheral OVA challenge, unlike mice given multiple nasal OVA doses that rendered them responsive to OVA. The observed unresponsiveness to OVA psigma1 could be adoptively transferred using cervical lymph node CD4(+) T cells, which failed to undergo proliferative or delayed-type hypersensitivity responses in recipients. To discern the cytokines responsible as a mechanism for this unresponsiveness, restimulation assays revealed increased production of regulatory cytokines, IL-4, IL-10, and TGF-beta1, with greatly reduced IL-17 and IFN-gamma. The induced IL-10 was derived predominantly from FoxP3(+)CD25(+)CD4(+) T cells. No FoxP3(+)CD25(+)CD4(+) T cells were induced in OVA-psigma1-dosed IL-10 deficient (IL-10(-/-)) mice, and despite showing increased TGF-beta1 synthesis, these mice were responsive to OVA. These data demonstrate the feasibility of using psigma1 as a mucosal delivery platform specifically for low-dose tolerance induction. PMID- 18390702 TI - 1alpha,25-dihydroxyvitamin D3 potentiates the beneficial effects of allergen immunotherapy in a mouse model of allergic asthma: role for IL-10 and TGF-beta. AB - 1alpha,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), a potent inhibitor of NF kappaB expression, can prevent the maturation of dendritic cells in vitro leading to tolerogenic dendritic cells with increased potential to induce regulatory T cells. Herein, we investigated whether the combination of allergen immunotherapy with 1,25(OH)(2)D(3) potentiates the suppressive effects of immunotherapy and whether the immunoregulatory cytokines IL-10 and TGF-beta are involved in the effector phase. OVA-sensitized and challenged BALB/c mice displayed airway hyperresponsiveness (AHR) and increased serum OVA-specific IgE levels, bronchoalveolar lavage eosinophilia, and Th2 cytokine levels. In this model, the dose response of allergen immunotherapy 10 days before OVA inhalation challenge shows strong suppression of asthma manifestations at 1 mg of OVA, but partial suppression of bronchoalveolar lavage eosinophilia, IgE up-regulation, and no reduction of AHR at 100 microg. Interestingly, coadministration of 10 ng of 1,25(OH)(2)D(3) with 100 microg of OVA immunotherapy significantly inhibited AHR and potentiated the reduction of serum OVA-specific IgE levels, airway eosinophilia, and Th2-related cytokines concomitant with increased IL-10 levels in lung tissues and TGF-beta and OVA-specific IgA levels in serum. Similar effects on suboptimal immunotherapy were observed by inhibition of the NF-kappaB pathway using the selective IkappaB kinase 2 inhibitor PS-1145. The suppressive effects of this combined immunotherapy were partially reversed by treatment with mAb to either IL-10R or TGF-beta before OVA inhalation challenge but completely abrogated when both Abs were given. These data demonstrate that 1,25(OH)(2)D(3) potentiates the efficacy of immunotherapy and that the regulatory cytokines IL-10 and TGF-beta play a crucial role in the effector phase of this mouse model. PMID- 18390704 TI - Differential contributions of APC subsets to T cell activation in nonobese diabetic mice. AB - Despite the pivotal role of dendritic cells (DC) in shaping immunity, little is known about their functionality in type 1 diabetes. Moreover, due to the paucity of DC in vivo, functional studies have relied largely upon in vitro-expanded cells to elucidate type 1 diabetes-associated functional abnormalities. In this study, we provide a comprehensive analysis of the functional capabilities of in vivo-derived DC subsets from NOD mice by comparing DC to other NOD APC types and to DC from autoimmune-resistant strains. NOD DC closely resemble those from nonautoimmune strains with respect to costimulation and cytokine production. The exception is the CD8alpha(+)CD11b(-)DC subset which is numerically reduced in NOD spleens, but not in the pancreatic lymph nodes, while DC from both tissues produce little IL-12 in this strain. This defect results in unusual deferral toward macrophage-derived IL-12 in NOD mice; NOD macrophages produce aberrantly high IL-12 levels that can overcompensate for the DC defect in Th1 polarization. APC subset use for autoantigen presentation also differs in NOD mice. NOD B cells overshadow DC at activating islet-reactive T cells, whereas DC and B cells in NOD resistant mice are functionally comparable. Differential involvement of APC subsets in T cell activation and tolerance induction may prove to be a crucial factor in the selection and expansion of autoreactive T cells. PMID- 18390703 TI - Down-regulation of ICOS ligand by interaction with ICOS functions as a regulatory mechanism for immune responses. AB - Although it is well-known that the ICOS-ICOS ligand (ICOSL) costimulatory pathway is important for many immune responses, recent accumulated evidence suggests that dysregulation of this pathway may lead to and/or exaggerate autoimmune responses. ICOS is induced on the cell surface after T cell activation. Similarly, ICOSL is up-regulated on APCs by several mitogenic stimuli. However, the mechanism regulating expression of the ICOS-ICOSL pair, and the significance of controlling their expression for an appropriate immune response, is largely unknown. To gain a better understanding of the importance of fine control of the ICOS-ICOSL costimulatory pathway, we generated ICOS-transgenic (Tg) mice that have high constitutive expression of ICOS in all T cells. Using ICOS-Tg mice, we studied whether in vivo immune responses were affected. Unexpectedly, we first found that ICOS-Tg mice exhibited a phenotype resembling ICOS-deficient mice in their Ag specific Ab response, such as a defect in class switch recombination. Further examination revealed that ICOSL expression of APCs was significantly suppressed in ICOS-Tg mice. Interestingly, suppression of ICOSL was induced by interaction of ICOSL with ICOS, and it seemed to be regulated at the posttranscriptional level. The suppressive effect of the ICOS-ICOSL interaction overcame the positive effect of CD40 or B cell activation factor of the TNF family (BAFF) stimulation on ICOSL expression. Together, our studies demonstrate a novel mechanism for the regulation of ICOSL expression in vivo and suggest that the ICOS costimulatory pathway is subject to negative feedback regulation by ICOSL down-regulation in response to ICOS expression. PMID- 18390705 TI - Neutralizing type I IFN antibodies trigger an IFN-like response in endothelial cells. AB - Neutralizing Abs to type I IFNs are of therapeutic significance, i.e., are currently evaluated for the treatment of autoimmune diseases with pathogenic IFN alpha production such as for systemic lupus erythematosus. Unexpectedly, we observed that several neutralizing Abs reportedly known to counteract IFN-alpha or IFN-beta activity triggered an "IFN-like" response in quiescent primary human endothelial cells leading to activation of the transcription factor IFN stimulated gene factor 3 and the expression of IFN-responsive genes. Furthermore, these Abs were found to enhance rather than inhibit type I IFN signals, and the effect was also detectable for distinct other cell types such as PBMCs. The stimulatory capacity of anti-IFN-alpha/beta Abs was mediated by the constitutive autocrine production of "subthreshold" IFN levels, involved the type I IFNR and was dependent on the Fc Ab domain, as Fab or F(ab')(2) fragments potently inhibited IFN activity. We thus propose that a combined effect of IFN recognition by the Ab paratope and the concomitant engagement of the Fc domain may trigger an IFN signal via the respective type I IFNR, which accounts for the observed IFN like response to the neutralizing Abs. With respect to clinical applications, the finding may be of importance for the design of recombinant Abs vs Fab or F(ab')(2) fragments to efficiently counteract IFN activity without undesirable activating effects. PMID- 18390706 TI - Polarization of primary human monocytes by IFN-gamma induces chromatin changes and recruits RNA Pol II to the TNF-alpha promoter. AB - Monocyte polarization by IFN-gamma or IL-4 drives a complex series of cellular responses leading to increased intracellular killing (IFN-gamma) or enhanced healing (IL-4) among other functional responses. We studied the effect of IL-4 and IFN-gamma polarization on histone modifications at the TNF-alpha locus in human primary monocytes. IFN-gamma polarization markedly increased the expression of TNF-alpha, whereas IL-4 treatment decreased the expression. We found that IFN gamma alone increased histone H4 acetylation at the TNF-alpha promoter. The effect of IFN-gamma on TNF-alpha expression was durable upon cytokine washout and even repolarization with IL-4. Concordantly, IFN-gamma-mediated H4 acetylation was also durable. IFN-gamma recruited activating transcription factor-2 via p38 to the TNF-alpha promoter, but inhibition of p38 had minimal effect on H4 acetylation. In a novel finding, we found that IFN-gamma recruited RNA Pol II to the human TNF-alpha promoter via ERK signaling, but did so without initiating transcription, leading to a poised condition. These studies provide an important perspective on monocyte polarization. Polarization by IFN-gamma has a durable effect on TNF-alpha expression, and histone acetylation may provide a mechanism for persistence of the effect. PMID- 18390708 TI - The apoptotic pathway contributing to the deletion of naive CD8 T cells during the induction of peripheral tolerance to a cross-presented self-antigen. AB - The maintenance of T cell tolerance in the periphery proceeds through several mechanisms, including anergy, immuno-regulation, and deletion via apoptosis. We examined the mechanism underlying the induction of CD8 T cell peripheral tolerance to a self-Ag expressed on pancreatic islet beta-cells. Following adoptive transfer, Ag-specific clone 4 T cells underwent deletion independently of extrinsic death receptors, including Fas, TNFR1, or TNFR2. Additional experiments revealed that the induction of clone 4 T cell apoptosis during peripheral tolerance occurred via an intrinsic death pathway that could be inhibited by overexpression of Bcl-2 or targeted deletion of the proapoptotic molecule, Bim, thereby resulting in accumulation of activated clone 4 T cells. Over-expression of Bcl-2 in clone 4 T cells promoted the development of effector function and insulitis whereas Bim-/- clone 4 cells were not autoaggressive. Examination of the upstream molecular mechanisms contributing to clone 4 T cell apoptosis revealed that it proceeded in a p53, E2F1, and E2F2-independent manner. Taken together, these data reveal that initiation of clone 4 T cell apoptosis during the induction of peripheral tolerance to a cross-presented self-Ag occurs through a Bcl-2-sensitive and at least partially Bim-dependent mechanism. PMID- 18390707 TI - CD137 costimulation of CD8+ T cells confers resistance to suppression by virus induced regulatory T cells. AB - Chronic viral infections cause high levels of morbidity and mortality worldwide, making the development of effective therapies a high priority for improving human health. We have used mice infected with Friend virus as a model to study immunotherapeutic approaches to the cure of chronic retroviral infections. In chronic Friend virus infections CD4(+) T regulatory (Treg) cells suppress CD8(+) T cell effector functions critical for virus clearance. In this study, we demonstrate that immunotherapy with a combination of agonistic anti-CD137 Ab and virus-specific, TCR-transgenic CD8(+) T cells produced greater than 99% reductions of virus levels within 2 wk. In vitro studies indicated that the CD137 specific Ab rendered the CD8(+) T cells resistant to Treg cell-mediated suppression with no direct effect on the suppressive function of the Treg cells. By 2 weeks after transfer, the adoptively transferred CD8(+) T cells were lost, likely due to activation-induced cell death. The highly focused immunological pressure placed on the virus by the single specificity CD8(+) T cells led to the appearance of escape variants, indicating that broader epitope specificity will be required for long-term virus control. However, the results demonstrate a potent strategy to potentiate the function of CD8(+) T cells in the context of immunosuppressive Treg cells. PMID- 18390709 TI - Aging down-regulates the transcription factor E2A, activation-induced cytidine deaminase, and Ig class switch in human B cells. AB - Elderly humans have compromised humoral and cellular immune responses, which lead to reduced protection to infectious agents and to vaccines. Currently, available vaccines suboptimally protect the elderly population. The capacity to class switch the Ig H chain is critical to the effectiveness of humoral immune responses in mice and humans. We have previously shown in mice that the E2A encoded transcription factor E47, which regulates many B cell functions, is down regulated in old splenic B cells. This leads to a reduction in the activation induced cytidine deaminase (AID), which is known to induce class switch recombination and Ig somatic hypermutation. The old activated murine B cells also have less AID and less switched Abs. We have extended our study here to investigate whether aging also affects Ab production and E47 and AID expression in B cells isolated from the peripheral blood of human subjects (18-86 years). Our results obtained with activated CD19(+) B cells show that the expression of E47, AID, and Iggamma1 circle transcripts progressively decrease with age. We also show an age-related decline in the percentage of switch memory B cells (IgG(+)/IgA(+)), an increase in that of naive B cells (IgG(-)/IgA(-)/CD27(-)) for most individuals, and no decrease in that of IgM memory cells in peripheral blood, consistent with our data on the decrease seen in class switch recombination in vitro. Our results provide a possible molecular mechanism for a B cell intrinsic defect in the humoral immune response with aging and suggest avenues for improvement of vaccine response in elderly humans. PMID- 18390710 TI - MyD88-dependent pathway in T cells directly modulates the expansion of colitogenic CD4+ T cells in chronic colitis. AB - TLRs that mediate the recognition of pathogen-associated molecular patterns are widely expressed on/in cells of the innate immune system. However, recent findings demonstrate that certain TLRs are also expressed in conventional TCRalphabeta(+) T cells that are critically involved in the acquired immune system, suggesting that TLR ligands can directly modulate T cell function in addition to various innate immune cells. In this study, we report that in a murine model of chronic colitis induced in RAG-2(-/-) mice by adoptive transfer of CD4(+)CD45RB(high) T cells, both CD4(+)CD45RB(high) donor cells and the expanding colitogenic lamina propria CD4(+)CD44(high) memory cells expresses a wide variety of TLRs along with MyD88, a key adaptor molecule required for signal transduction through TLRs. Although RAG-2(-/-) mice transferred with MyD88(-/ )CD4(+)CD45RB(high) cells developed colitis, the severity was reduced with the delayed kinetics of clinical course, and the expansion of colitogenic CD4(+) T cells was significantly impaired as compared with control mice transferred with MyD88(+/+)CD4(+)CD45RB(high) cells. When RAG-2(-/-) mice were transferred with the same number of MyD88(+/+) (Ly5.1(+)) and MyD88(-/-) (Ly5.2(+)) CD4(+)CD45RB(high) cells, MyD88(-/-)CD4(+) T cells showed significantly lower proliferative responses assessed by in vivo CFSE division assay, and also lower expression of antiapoptotic Bcl-2/Bcl-x(L) molecules and less production of IFN gamma and IL-17, compared with the paired MyD88(+/+)CD4(+) T cells. Collectively, the MyD88-dependent pathway that controls TLR signaling in T cells may directly promote the proliferation and survival of colitogenic CD4(+) T cells to sustain chronic colitis. PMID- 18390711 TI - Derivation and fluidity of acutely induced dysfunctional CD8+ T cells. AB - Dysfunctional CD8(+) T (T(CD8(+))) cells lacking cytokine production have been identified in many viral infections, but their genesis is not well understood. Established results indicate that such cells could be either high avidity that enter a refractory state due to overstimulation or low avidity that are only partially stimulated. Using an acute, resolving infection model that results in rapid production of dysfunctional cells, we show that this IL2 unresponsive phenotype emerges from the low end of the avidity spectrum and is characterized by broad TCR usage and a reduced proliferation rate. Furthermore, the dysfunctional population is extremely fluid, being sustained by high Ag dose but virtually eliminated following low dose boosting. Together, these results suggest that persistence of dysfunctional cells generated in this manner depends upon continual exposure to high Ag levels and that such cells may ultimately predominate if functional cells become exhausted. PMID- 18390713 TI - The lymphopenic environment of CD132 (common gamma-chain)-deficient hosts elicits rapid homeostatic proliferation of naive T cells via IL-15. AB - Homeostatic proliferation for naive T cells is observed readily only under lymphopenic conditions in response to elevated levels of IL-7 and contact with self-MHC/peptide ligands. Homeostatic proliferation occurs at a slow pace and gradually induces the dividing cells to acquire characteristics of memory cells. We describe a novel type of homeostatic proliferation whereby naive T cells proliferate at a significantly faster rate, resembling the proliferation speed induced by foreign Ags, and the expanding cells rapidly differentiate into central memory cells. Remarkably, such rapid homeostatic proliferation is driven by a combination of IL-2 and IL-15, with IL-15 playing a bigger role, and applies for a wide repertoire of CD8(+) naive T cells, including many TCR-transgenic lines, even those that fail to undergo IL-7-driven homeostatic proliferation. Thus, naive T cells can be induced to undergo homeostatic proliferation of variable speed with a few members of the common gamma-chain (CD132) family of cytokines, the speed of proliferation depending on the levels of the particular cytokine involved. PMID- 18390712 TI - Formation of IL-7Ralphahigh and IL-7Ralphalow CD8 T cells during infection is regulated by the opposing functions of GABPalpha and Gfi-1. AB - IL-7 is essential for the survival of naive and memory T cells, and IL-7 receptor alpha-chain (IL-7Ralpha) expression is dynamically regulated in activated CD8 T cells during acute viral and bacterial infections. Most virus-specific CD8 T cells become IL-7Ralpha(low) and are relatively short-lived, but some escape IL 7Ralpha repression (referred to as IL-7Ralpha(high) memory precursor effector cells) and preferentially enter the memory CD8 T cell pool. How antiviral effector CD8 T cells regulate IL-7Ralpha expression in an "on and off" fashion remains to be characterized. During lymphocytic choriomeningitis virus infection, we found that opposing actions of the transcription factors GABPalpha (GA binding protein alpha) and Gfi-1 (growth factor independence 1) control IL-7Ralpha expression in effector CD8 T cells. Specifically, GABPalpha was required for IL 7Ralpha expression in memory precursor effector cells, and this correlated with hyperacetylation of the Il7ra promoter. In contrast, Gfi-1 was required for stable IL-7Ralpha repression in effector CD8 T cells and acted by antagonizing GABPalpha binding and recruiting histone deacetylase 1, which deacetylated the Il7ra promoter. Thus, Il7ra promoter acetylation and activity was dependent on the reciprocal binding of GABPalpha and Gfi-1, and these data provide a biochemical mechanism for the generation of stable IL-7Ralpha(high) and IL 7Ralpha(low) states in virus-specific effector CD8 T cells. PMID- 18390714 TI - Effector memory alphabeta T lymphocytes can express FcgammaRIIIa and mediate antibody-dependent cellular cytotoxicity. AB - Human memory T cells are comprised of distinct populations with different homing potential and effector functions: central memory T cells that mount recall responses to Ags in secondary lymphoid organs, and effector memory T cells that confer immediate protection in peripheral tissues. In the present study we demonstrate that a proportion of effector memory T cells express FcgammaRIIIa (CD16), are perforin positive, and directly mediate Ab-dependent cytotoxicity ex vivo. This particular alphabeta T lymphocyte subset has the morphology of large granular lymphocytes, increases proportionately in vivo during reactive lymphocytosis, and can be detected in vitro among EBV-specific T lymphocytes after stimulation with EBV Ags. Consequently, during a normal immune response, amplification of these effector memory T lymphocytes that are capable of Ab dependent cytotoxicity may have beneficial or harmful consequences depending on the presence of pathogen- or tissue-specific Abs, respectively. PMID- 18390715 TI - Sphingosine 1-phosphate regulates the egress of IgA plasmablasts from Peyer's patches for intestinal IgA responses. AB - It is well established that Peyer's patches (PPs) are sites for the differentiation of IgA plasma cell precursors, but molecular and cellular mechanisms in their trafficking remain to be elucidated. In this study, we show that alterations in type 1 sphingosine 1-phosphate (S1P) receptor expression during B cell differentiation in the PPs control the emigration of IgA plasma cell precursors. Type 1 S1P receptor expression decreased during the differentiation of IgM(+)B220(+) B cells to IgA(+)B220(+) B cells, but recovered on IgA(+)B220(-) plasmablasts for their emigration from the PPs. Thus, IgA(+)B220(-) plasmablasts migrated in response to S1P in vitro. Additionally, IgA(+) plasmablasts selectively accumulated in lymphatic regions of PPs when S1P mediated signaling was disrupted by FTY720 treatment. This accumulation of IgA(+) plasmablasts in the PPs led to their reduction in the intestinal lamina propria and simultaneous impairment of Ag-specific intestinal IgA production against orally administered Ag. These findings suggest that S1P regulates the retention and emigration of PP B cells and plays key roles in the induction of intestinal IgA production. PMID- 18390716 TI - Neural crest origin of perivascular mesenchyme in the adult thymus. AB - The endodermal epithelial thymus anlage develops in tight association with neural crest (NC)-derived mesenchyme. This epithelial-NC interaction is crucial for thymus development, but it is not known how NC supports thymus development or whether NC cells or their progeny make any significant contribution to the adult thymus. By nude mouse blastocyst complementation and by cell surface phenotype, we could previously separate thymus stroma into Foxn1-dependent epithelial cells and a Foxn1-independent mesenchymal cell population. These mesenchymal cells expressed vascular endothelial growth factor-A, and contributed to thymus vascularization. These data suggested a physical or functional association with thymic blood vessels, but the origin, location in the thymus, and function of these stromal cells remained unknown. Using a transgenic mouse expressing Cre recombinase in premigratory NC (Sox10-Cre), we have now fate-mapped the majority of these adult mesenchymal cells to a NC origin. NC-derived cells represent tightly vessel-associated pericytes that are sandwiched between endothelium and epithelium along the entire thymus vasculature. The ontogenetic, phenotypic, and positional definition of this distinct perivascular mesenchymal compartment provides a cellular basis for the role of NC in thymus development and possibly maintenance, and might be useful to address properties of the endothelial epithelial barrier in the adult thymus. PMID- 18390717 TI - Inter-strain tissue-infiltrating T cell responses to minor histocompatibility antigens involved in graft-versus-host disease as determined by Vbeta spectratype analysis. AB - Lethal graft-vs-host disease (GVHD) can be induced between MHC-matched murine strains expressing multiple minor histocompatibility Ag differences. In the B6 >BALB.B model, both CD4(+) and CD8(+) donor T cells can mediate lethal GVHD, whereas in the B6->CXB-2 model, only CD8(+) T cells are lethal. TCR Vbeta CDR3 size spectratyping was previously used to analyze CD8(+) and CD4(+) T cell responses in lethally irradiated BALB.B and CXB-2 recipients, which showed significant overlap in the reacting repertoires. However, CD4(+) T cells exhibited unique skewing of the Vbeta2 and 11 families in only BALB.B recipients. These Vbeta family reactivities were confirmed by immunohistochemical staining of lingual epithelial infiltrates, and by positive and negative selection Vbeta family transfer experiments for GVHD induction in BALB.B recipients. We have now extended these studies to examine the T cell repertoire responses involved in target tissue damage. Infiltrating B6 host-presensitized CD8(+) and CD4(+) T cells were isolated 8-10 days post-transplant from the spleens, intestines and livers of CXB-2 and BALB.B transplant recipients. For both T cell subsets, the results indicated overlapping tissue skewings between the recipients, also between the tissues sampled within the respective recipients as well as tissue specific responses unique to both the BALB.B and CXB-2 infiltrates. Most notably, the CD4(+) Vbeta 11(+) family was skewed in the intestines of BALB.B but not CXB 2 recipients. Taken together, these data suggest that there are likely to be target tissue-related anti-multiple minor histocompatibility Ag-specific responses in each of the strain recipients, which may also differ from those found in peripheral lymphoid organs. PMID- 18390718 TI - Induction of adaptive T regulatory cells that suppress the allergic response by coimmunization of DNA and protein vaccines. AB - Allergen-induced immediate hypersensitivity (AIH) is a health issue of significant concern. This robust inflammatory reaction is initiated by the allergen-specific T cell responsiveness. Severe lesion reactions on skin are consequential problem requiring medical treatment. Effective Ag-specific treatments or preventions are lacking. Using a rodent model of AIH induced by flea allergens, we first report that coimmunization of DNA and protein vaccines encoding the flea salivary specific Ag-1 ameliorated experimental AIH, including Ag-induced wheal formation, elevated T cell proliferation, and infiltration of lymphocytes and mast cells to the site of allergen challenge. The amelioration of AIH was directly related to the induction of a specific population of flea antigenic specific T cells exhibiting a CD4(+)CD25(-)FoxP3(+) phenotype, a characteristic of regulatory T (T(REG)) cells. These T(REG) cells expressing IL 10, IFN-gamma, and the transcriptional factor T-bet after Ag stimulation were driven by a tolerogenic MHC class II(+)/CD40(low) dendritic cell population that was induced by the coimmunization of DNA and protein vaccines. The tolerogenic dendritic cell could educate the naive T cells into CD4(+)CD25(-)FoxP3(+) T(REG) cells both in vitro and in vivo. The study identified phenomenon to induce an Ag specific tolerance via a defined Ag vaccinations and lead to the control of AIH. Exploitation of these cellular regulators and understanding their induction provides a basis for the possible development of novel therapies against allergic and related disorders in humans and animals. PMID- 18390719 TI - Zebrafish TRIF, a Golgi-localized protein, participates in IFN induction and NF kappaB activation. AB - The antiviral immune responses were triggered by the innate immune recognition of viral infection. The type I IFNs (IFN-beta and IFN-alpha) are the key cytokines produced upon viral infection and consequently link innate immunity with adaptive immunity. A main antiviral system in mammals is TRIF-dependent TLRs pathway, but the TRIF-independent RIG-I pathway, has also been discovered recently. In this manuscript, our study focuses on the functional characterization of zebrafish TRIF based on the comparison of its sequence and functional evolution from zebrafish to mammals. Our experimental results show that the full length cDNA of zebrafish TRIF cloned by RACE-PCR approach encodes a protein of 556 amino acids. Luciferase reporter assay confirms that zebrafish TRIF is able to induce the IFN promoter as well as activate NF-kappaB response promoter. The IFN induction function of zebrafish TRIF is abolished when Ala359 is mutated to Pro or His. Laser confocal microscopy shows that zebrafish TRIF is colocalized with a Golgi apparatus marker, implying its unique subcellular localization in Golgi apparatus. In zebrafish, the mRNA expression of molecules participating in RIG-I pathway are much more sensitive and specific to polyinosine-polycytidylic acid induction compared with those in TRIF-dependent antiviral pathway. The TRIF dependent TLR4 IFN induction signaling appears not to be functional in zebrafish, since IFN expression cannot be up-regulated by LPS. These two striking findings from de novo ligand induction experiments suggest a novel antiviral mechanism in zebrafish. PMID- 18390720 TI - The lymphotoxin pathway regulates Aire-independent expression of ectopic genes and chemokines in thymic stromal cells. AB - Medullary thymic epithelial cells (mTEC) play an important and unique role in central tolerance, expressing tissue-restricted Ags (TRA) which delete thymocytes autoreactive to peripheral organs. Since deficiencies in this cell type or activity can lead to devastating autoimmune diseases, it is important to understand the factors which regulate mTEC differentiation and function. Lymphotoxin (LT) ligands and the LTbetaR have been recently shown to be important regulators of mTEC biology; however, the precise role of this pathway in the thymus is not clear. In this study, we have investigated the impact of this signaling pathway in greater detail, focusing not only on mTEC but also on other thymic stromal cell subsets. LTbetaR expression was found in all TEC subsets, but the highest levels were detected in MTS-15(+) thymic fibroblasts. Rather than directing the expression of the autoimmune regulator Aire in mTEC, we found LTbetaR signals were important for TRA expression in a distinct population of mTEC characterized by low levels of MHC class II (mTEC(low)), as well as maintenance of MTS-15(+) fibroblasts. In addition, thymic stromal cell subsets from LT-deficient mice exhibit defects in chemokine production similar to that found in peripheral lymphoid organs of Lta(-/-) and Ltbr(-/-) mice. Thus, we propose a broader role for LTalpha1beta2-LTbetaR signaling in the maintenance of the thymic microenvironments, specifically by regulating TRA and chemokine expression in mTEC(low) for efficient induction of central tolerance. PMID- 18390721 TI - Release from regulatory T cell-mediated suppression during the onset of tissue specific autoimmunity is associated with elevated IL-21. AB - The activity of regulatory T cells (Treg) is widely accepted to play a central role in preventing pathogenic immune responses against self-Ags. However, it is not clear why such regulation breaks down during the onset of autoimmunity. We have studied self-Ag-specific Treg during the induction of spontaneous diabetes. Our data reveal a shift in the balance between regulatory and pathogenic islet reactive T cells in the pancreas-draining lymph nodes during disease onset. Treg function was not compromised during disease initiation, but instead conventional T cells showed reduced susceptibility to Treg-mediated suppression. Release from Treg suppression was associated with elevated levels of IL-21 in vivo, and provision of this cytokine abrogated Treg suppression in vitro and in vivo. These data suggest that immunological protection of a peripheral tissue by Treg can be subverted by IL-21, suggesting new strategies for intervention in autoimmunity. PMID- 18390722 TI - The adjuvants aluminum hydroxide and MF59 induce monocyte and granulocyte chemoattractants and enhance monocyte differentiation toward dendritic cells. AB - Aluminum hydroxide (alum) and the oil-in-water emulsion MF59 are widely used, safe and effective adjuvants, yet their mechanism of action is poorly understood. We assessed the effects of alum and MF59 on human immune cells and found that both induce secretion of chemokines, such as CCL2 (MCP-1), CCL3 (MIP-1alpha), CCL4 (MIP-1beta), and CXCL8 (IL-8), all involved in cell recruitment from blood into peripheral tissue. Alum appears to act mainly on macrophages and monocytes, whereas MF59 additionally targets granulocytes. Accordingly, monocytes and granulocytes migrate toward MF59-conditioned culture supernatants. In monocytes, both adjuvants lead to increased endocytosis, enhanced surface expression of MHC class II and CD86, and down-regulation of the monocyte marker CD14, which are all phenotypic changes consistent with a differentiation toward dendritic cells (DCs). When monocyte differentiation into DCs is induced by addition of cytokines, these adjuvants enhanced the acquisition of a mature DC phenotype and lead to an earlier and higher expression of MHC class II and CD86. In addition, MF59 induces further up-regulation of the maturation marker CD83 and the lymph node-homing receptor CCR7 on differentiating monocytes. Alum induces a similar but not identical pattern that clearly differs from the response to LPS. This model suggests a common adjuvant mechanism that is distinct from that mediated by danger signals. We conclude that during vaccination, adjuvants such as MF59 may increase recruitment of immune cells into the injection site, accelerate and enhance monocyte differentiation into DCs, augment Ag uptake, and facilitate migration of DCs into tissue-draining lymph nodes to prime adaptive immune responses. PMID- 18390723 TI - Pirk is a negative regulator of the Drosophila Imd pathway. AB - NF-kappaB transcription factors are involved in evolutionarily conserved signaling pathways controlling multiple cellular processes including apoptosis and immune and inflammatory responses. Immune response of the fruit fly Drosophila melanogaster to Gram-negative bacteria is primarily mediated via the Imd (immune deficiency) pathway, which closely resembles the mammalian TNFR signaling pathway. Instead of cytokines, the main outcome of Imd signaling is the production of antimicrobial peptides. The pathway activity is delicately regulated. Although many of the Imd pathway components are known, the mechanisms of negative regulation are more elusive. In this study we report that a previously uncharacterized gene, pirk, is highly induced upon Gram-negative bacterial infection in Drosophila in vitro and in vivo. pirk encodes a cytoplasmic protein that coimmunoprecipitates with Imd and the cytoplasmic tail of peptidoglycan recognition protein LC (PGRP-LC). RNA interference-mediated down regulation of Pirk caused Imd pathway hyperactivation upon infection with Gram negative bacteria, while overexpression of pirk reduced the Imd pathway response both in vitro and in vivo. Furthermore, pirk-overexpressing flies were more susceptible to Gram-negative bacterial infection than wild-type flies. We conclude that Pirk is a negative regulator of the Imd pathway. PMID- 18390724 TI - Abrogation of anti-retinal autoimmunity in IL-10 transgenic mice due to reduced T cell priming and inhibition of disease effector mechanisms. AB - Experimental autoimmune uveitis (EAU) induced by immunization of animals with retinal Ags is a model for human uveitis. The immunosuppressive cytokine IL-10 regulates EAU susceptibility and may be a factor in genetic resistance to EAU. To further elucidate the regulatory role of endogenous IL-10 in the mouse model of EAU, we examined transgenic (Tg) mice expressing IL-10 either in activated T cells (inducible) or in macrophages (constitutive). These IL-10-Tg mice and non Tg wild-type controls were immunized with a uveitogenic regimen of the retinal Ag interphotoreceptor retinoid-binding protein. Constitutive expression of IL-10 in macrophages abrogated disease and reduced Ag-specific immunological responses. These mice had detectable levels of IL-10 in sera and in ocular extracts. In contrast, expression of IL-10 in activated T cells only partially protected from EAU and marginally reduced Ag-specific responses. All IL-10-Tg lines showed suppression of Ag-specific effector cytokines. APC from Tg mice constitutively expressing IL-10 in macrophages exhibited decreased ability to prime naive T cells, however, Ag presentation to already primed T cells was not compromised. Importantly, IL-10-Tg mice that received interphotoreceptor retinoid-binding protein-specific uveitogenic T cells from wild-type donors were protected from EAU. We suggest that constitutively produced endogenous IL-10 ameliorates the development of EAU by suppressing de novo priming of Ag-specific T cells and inhibiting the recruitment and/or function of inflammatory leukocytes, rather than by inhibiting local Ag presentation within the eye. PMID- 18390725 TI - Equivalent specificity of peripheral blood and islet-infiltrating CD8+ T lymphocytes in spontaneously diabetic HLA-A2 transgenic NOD mice. AB - CD8(+) T cells play an important role in the initiation of insulitis and in the destructive stage leading to insulin-dependent diabetes mellitus. A string of recent studies has led to the identification of numerous HLA-A2-restricted epitopes derived from pancreatic beta cell Ags. It is hoped that assays detecting responses of patient PBMC to such epitopes might be instrumental for early diagnosis of beta cell-directed autoimmunity and for monitoring trials of immunointervention. However, it remains unclear whether the results of assays studying PBMC reflect responses of islet-infiltrating lymphocytes, and to what extent they correlate with disease risk and/or activity. We have used female and male humanized NOD mice expressing HLA-A2 in addition to murine MHC class I molecules to study spontaneous responses of islet-infiltrating blood, spleen, and lymph node lymphocytes of various age groups to a panel of 16 epitopes. Twelve of these are restricted by HLA-A2, have previously been shown to be recognized by patient CTL, and have identical sequences in human and murine autoantigens. Using an IFN-gamma ELISPOT assay, we find highly similar hierarchies of epitope immunodominance in the different T cell compartments, including peripheral blood and pancreatic islets. Moreover, we demonstrate that most of the epitopes eliciting dominant responses in humans display similar status in the mouse model. These results emphasize the potential of humanized mice as tools for studying spontaneous autoimmune CTL responses, and they provide a strong rationale for the development and use of assays monitoring responses of CD8(+) PBMC in human type 1 diabetes. PMID- 18390726 TI - Immune activation driven by CTLA-4 blockade augments viral replication at mucosal sites in simian immunodeficiency virus infection. AB - The importance of chronic immune activation in progression to AIDS has been inferred by correlative studies in HIV-infected individuals and in nonhuman primate models of SIV infection. Using the SIV(mac251) macaque model, we directly address the impact of immune activation by inhibiting CTLA-4, an immunoregulatory molecule expressed on activated T cells and a subset of regulatory T cells. We found that CTLA-4 blockade significantly increased T cell activation and viral replication in primary SIV(mac251) infection, particularly at mucosal sites, and increased IDO expression and activity. Accordingly, protracted treatment with anti-CTLA-4 Ab of macaques chronically infected with SIV(mac251) decreased responsiveness to antiretroviral therapy and abrogated the ability of therapeutic T cell vaccines to decrease viral set point. These data provide the first direct evidence that immune activation drives viral replication, and suggest caution in the use of therapeutic approaches for HIV infection in vivo that increase CD4(+) T cell proliferation. PMID- 18390727 TI - ICOS/ICOSL interaction is required for CD4+ invariant NKT cell function and homeostatic survival. AB - The development of airway hyperreactivity (AHR), a cardinal feature of asthma, requires the presence of invariant NKT (iNKT) cells. In a mouse model of asthma, we demonstrated that the induction of AHR required ICOS costimulation of iNKT cells. ICOS was highly expressed on both naive and activated iNKT cells, and expression of ICOS was greater on the CD4(+) iNKT than on CD4(-) iNKT cells. Furthermore, the number of CD4(+) iNKT cells was significantly lower in spleens and livers of ICOS(-/-) and ICOSL(-/-) mice, and the remaining iNKT cells in ICOS(-/-) mice were dysfunctional and failed to reconstitute AHR when adoptively transferred into iNKT cell-deficient Jalpha18(-/-) mice. In addition, direct activation of iNKT cells with alpha-GalCer, which induced AHR in wild-type mice, failed to induce AHR in ICOS(-/-) mice. The failure of ICOS(-/-) iNKT cells to induce AHR was due in part to an inability of the ICOS(-/-) iNKT cells to produce IL-4 and IL-13 on activation. Moreover, survival of wild-type iNKT cells transferred into ICOSL(-/-) mice was greatly reduced due to the induction of apoptosis. These results indicate that ICOS costimulation plays a major role in induction of AHR by iNKT cells and is required for CD4(+) iNKT cell function, homeostasis, and survival in the periphery. PMID- 18390728 TI - Nonoverlapping expression of IL10, IL12p40, and IFNgamma mRNA in the marginal zone and T cell zone of the spleen after antigenic stimulation. AB - The differentiation of CD4(+) T cells is regulated by cytokines locally within the compartments of secondary lymphoid organs during adaptive immune responses. Quantitative data about the expression of cytokine mRNAs within the T and B cell zones of lymphoid organs are lacking. In this study, we assessed the expression of multiple cytokine genes within the lymphoid compartments of the spleen of rats after two types of stimulation. First, the spleen was stimulated directly by a blood-derived Ag. Second, the spleen was stimulated indirectly by incoming lymphocytes that had been activated and released during a proceeding immune response at a distant tissue site. Using laser microdissection, we show that the expression of cytokine mRNAs was compartment specific, transient, and preceded cell proliferation after the direct antigenic stimulation. Surprisingly, the indirect stimulation by incoming activated lymphocytes induced similar cytokines in the T cell zone. However, the nonoverlapping expression was lost and IL10 appeared as the major cytokine in all compartments. Thus, tracking two types of immune activation without disturbing the integrity of structures reveals distinct and overlapping events in the compartments of the spleen. This information adds a new dimension to the understanding of immune responses in vivo. PMID- 18390729 TI - Unique effects of KIT D816V in BaF3 cells: induction of cluster formation, histamine synthesis, and early mast cell differentiation antigens. AB - Oncogenic tyrosine kinases (TK) usually convert growth factor-dependent cells to factor independence with autonomous proliferation. However, TK-driven neoplasms often are indolent and characterized by cell differentiation rather than proliferation. A prototype of an indolent TK-driven neoplasm is indolent systemic mastocytosis. We found that the D816V-mutated variant of KIT, a TK detectable in most patients with systemic mastocytosis, induces cluster formation and expression of several mast cell differentiation and adhesion Ags, including microphthalmia transcription factor, IL-4 receptor, histamine, CD63, and ICAM-1 in IL-3-dependent BaF3 cells. By contrast, wild-type KIT did not induce cluster formation or mast cell differentiation Ags. Additionally, KIT D816V, but not wild type KIT, induced STAT5 activation in BaF3 cells. However, despite these intriguing effects, KIT D816V did not convert BaF3 cells to factor-independent proliferation. Correspondingly, BaF3 cells with conditional expression of KIT D816V did not form tumors in nude mice. Together, the biologic effects of KIT D816V in BaF3 cells match strikingly with the clinical course of indolent systemic mastocytosis and with our recently established transgenic mouse model, in which KIT D816V induces indolent mast cell accumulations but usually does not induce a malignant mast cell disease. Based on all these results, it is hypothesized that KIT D816V as a single hit may be sufficient to cause indolent systemic mastocytosis, whereas additional defects may be required to induce aggressive mast cell disorders. PMID- 18390730 TI - The IL-1 family member 7b translocates to the nucleus and down-regulates proinflammatory cytokines. AB - The IL-1 family member 7b (IL-1F7b) is a novel homolog of the IL-1 cytokine family discovered by computational cloning. We have reported that IL-1F7b shares critical amino acid residues with IL-18 and binds the IL-18-binding protein; in doing so, IL-1F7b augments the inhibition of IFN-gamma by the IL-18-binding protein. IL-1F7b also binds IL-18Ralpha but neither induces signal nor acts as a receptor antagonist. Hence, the function of IL-1F7b remains unknown. In the present study, we analyzed the intracellular expression pattern of IL-1F7b. Using two variants of GFP fusion constructs of human IL-1F7b stably expressed in RAW macrophages, only the postcleavage mature form of the IL-1F7b precursor-but not the N-terminal propiece-specifically translocates to the nucleus following LPS stimulation. IL-1F7b, like IL-1beta, IL-18, and IL-33, is processed by caspase-1 to generate the mature cytokines. Therefore, we tested whether caspase-1-mediated cleavage of the IL-1F7b precursor is required for mature IL-1F7b to translocate actively into the nucleus. Indeed, a specific caspase-1 inhibitor markedly reduced nuclear entry of IL-1F7b. In stable transfectants of human IL-1F7b in RAW macrophages stimulated with LPS, levels of TNF-alpha, IL-1alpha, IL-6, as well as the chemokine MIP-2, were substantially reduced (72-98%) compared with LPS stimulated cells transfected with the empty plasmid. These results demonstrate that IL-1F7b translocates to the nucleus after caspase-1 processing and may act as a transcriptional modulator reducing the production of LPS-stimulated proinflammatory cytokines, consistent with IL-1F7b being an anti-inflammatory member of the IL-1 family. PMID- 18390731 TI - A conserved IFN-alpha receptor tyrosine motif directs the biological response to type I IFNs. AB - Mammalian type I IFNs (IFN-Is) mediate their potent biological activities through an evolutionarily conserved IFN-alpha receptor (IFNAR), consisting of IFNAR1 and IFNAR2. These two chains direct the rapid activation of two founding members of the STAT family of transcription factors, STAT1 and STAT2. To understand how IFN Is direct the recruitment and activation of STATs, a series of mutant murine IFNAR1 and IFNAR2 receptors were generated and evaluated in IFNAR1 and IFNAR2 knockout cells. These studies reveal that a single conserved IFNAR2 tyrosine, Y(510), plays a critical role in directing the IFN-I-dependent activation of STAT1 and STAT2, both in murine fibroblasts and macrophages. A second IFNAR2 tyrosine, Y(335), plays a more minor role. Likewise, Y(510) > Y(335) play a critical role in the induction of genes and antiviral activity traditionally associated with IFN-Is. PMID- 18390732 TI - Immunoproteomic identification and serological responses to novel Chlamydia pneumoniae antigens that are associated with persistent C. pneumoniae infections. AB - The controversial discussion about the role of Chlamydia pneumoniae in atherosclerosis cannot be solved without a reliable diagnosis that allows discrimination between past and persistent infections. Using a proteomic approach and immunoblotting with human sera, we identified 31 major C. pneumoniae Ags originating from 27 different C. pneumoniae proteins. More than half of the proteins represent Chlamydia Ags not described previously. Using a comparative analysis of spot reactivity Pmp6, OMP2, GroEL, DnaK, RpoA, EF-Tu, as well as CpB0704 and CpB0837, were found to be immunodominant. The comparison of Ab response patterns of sera from subjects with and without evidence for persisting C. pneumoniae, determined by multiple PCR analysis of PBMC and vasculatory samples, resulted in differential reactivity for 12 proteins, which is not reflected by reactivity of the sera in the microimmunofluorescence test, the current gold standard for serodiagnosis. Although reactivity of sera from PCR positive donors was increased toward RpoA, MOMP, YscC, Pmp10, PorB, Pmp21, GroEL, and Cpaf, the reactivity toward YscL, Rho, LCrE, and CpB0837 was decreased, reflecting the altered protein expression of persisting C. pneumoniae in vitro. Our data provide the first evidence of a unique Ab-response pattern associated with persistent C. pneumoniae infections, which is a prerequisite for the serological determination of persistently infected patients. PMID- 18390733 TI - Targeting the extracellular membrane-proximal domain of membrane-bound IgE by passive immunization blocks IgE synthesis in vivo. AB - The classical allergic reaction starts seconds or minutes after Ag contact and is committed by Abs produced by a special subset of B lymphocytes. These Abs belong to the IgE subclass and are responsible for Type I hyperreactivity reactions. Treatment of allergic diseases with humanized anti-IgE Abs leads primarily to a decrease of serum IgE levels. As a consequence, the number of high-affinity IgE receptors on mast cells and basophils decreases, leading to a lower excitability of the effector cells. The biological mechanism behind anti-IgE therapy remains partly speculative; however, it is likely that these Abs also interact with membrane IgE (mIgE) on B cells and possibly interfere with IgE production. In the present work, we raised a mouse mAb directed exclusively against the extracellular membrane-proximal domain of mIgE. The interaction between the monoclonal anti-mIgE Ab and mIgE induces receptor-mediated apoptosis in vitro. Passive immunization experiments lead to a block of newly synthesized specific IgEs during a parallel application of recombinant Bet v1a, the major birch pollen allergen. The decrease of allergen-specific serum IgE might be related to tolerance-inducing mechanisms stopping mIgE-displaying B cells in their proliferation and differentiation. PMID- 18390734 TI - The long isoform of cellular FLIP is essential for T lymphocyte proliferation through an NF-kappaB-independent pathway. AB - Although the long isoform of cellular FLIP (c-FLIP(L)) has been implicated in TCR mediated signaling, its role in T cell proliferation remains controversial. Some studies have demonstrated that overexpression of c-FLIP(L) promotes T cell proliferation and NF-kappaB activation, whereas others have reported that c FLIP(L) overexpression has no effect or even inhibits T cell proliferation. To establish the role of c-FLIP(L) in T lymphocyte proliferation, we have generated a conditional knockout mouse strain specifically lacking c-FLIP(L) in T lymphocytes. c-FLIP(L)(-/-) mice exhibit severely impaired effector T cell development after Listeria monocytogenes infection in vivo and c-FLIP(L) deficient T cells display defective TCR-mediated proliferation in vitro. However, c-FLIP(L)(-/-) T cells exhibit normal NF-kappaB activity upon TCR stimulation. These results demonstrate that c-FLIP(L) is essential for T lymphocyte proliferation through an NF-kappaB-independent pathway. PMID- 18390735 TI - Homeostatic intracellular-free Ca2+ is permissive for Rap1-mediated constitutive activation of alpha4 integrins on eosinophils. AB - Although much progress has been made in understanding the molecular mechanisms underlying agonist-induced "inside-out" activation of integrins, little is known about how basal levels of integrin function are maintained. This is particularly important for nonactivated eosinophils, where intermediate activation of alpha(4)beta(1) integrin supports recruitment to endothelial cells under flow conditions. Depletion of intracellular Ca(2+) and pharmacological inhibition of phospholipase C (but not other intracellular signaling molecules, including PI3K, ERK1/2, p38 MAPK, and tyrosine kinase activity) abrogated basal alpha(4) integrin activity in nonactivated eosinophils. Basal alpha(4) integrin activation was associated with activation of the small GTPase Rap1, a known regulator of agonist induced integrin function. Basal Rap activation was dependent upon phospholipase C, but not intracellular Ca(2+). However, depletion of intracellular Ca(2+) in CD34(+) hematopoietic progenitor cells abolished RapV12-mediated induction of alpha(4) integrin activity. Thus, residual Rap activity or constitutively active Rap activity in Ca(2+)-depleted cells is not sufficient to induce alpha(4) integrin activation. These data suggest that activation of functional alpha(4) integrin activity in resting eosinophils is mediated by Rap1 provided that the intracellular-free Ca(2+) is at a normal homeostatic concentration. PMID- 18390736 TI - A 9-centimorgan interval of chromosome 10 controls the T cell-dependent psoriasiform skin disease and arthritis in a murine psoriasis model. AB - Psoriasis is a complex genetic disease of unresolved pathogenesis with both heritable and environmental factors contributing to onset and severity. In addition to a disfiguring skin inflammation, approximately 10-40% of psoriasis patients suffer from destructive joint involvement. Previously, we reported that the CD18 hypomorphic PL/J mouse carrying a mutation resulting in reduced expression of the common chain of beta(2) integrins (CD11/CD18) spontaneously develops a skin disease that closely resembles human psoriasis. In contrast, the same mutation on C57BL/6J background did not demonstrate this phenotype. By a genome-wide linkage analysis, two major loci were identified as contributing to the development of psoriasiform dermatitis under the condition of low CD18 expression. Using a congenic approach, we now demonstrate that the introduction of a 9-centimorgan fragment of chromosome 10 derived from the PL/J strain into the disease-resistant CD18 hypomorphic C57BL/6J was promoting the development of psoriasiform skin disease and notably also arthritis. We therefore designated this locus psoriasiform skin disease-associated locus 1 (PSD1). High numbers of CD4(+) T cells and TNF-alpha producing macrophages were detected both in inflamed skin and joints in these congenic mice, with a complete resolution upon TNF-alpha inhibitor therapy or depletion of CD4(+) T cells. For the first time, we have identified a distinct genetic element that contributes to the T cell-dependent development of both psoriasiform skin disease and associated arthritis. This congenic model will be suitable to further investigations of genetic and molecular pathways that cause psoriasiform dermatitis and arthritis, and it may also be relevant for other autoimmune diseases. PMID- 18390737 TI - Chemokine and cytokine mediated loss of regulatory T cells in lymph nodes during pathogenic simian immunodeficiency virus infection. AB - Regulatory T cells (T(reg)) play key roles in immune regulation through multiple modes of suppression. The effects of HIV-1 infection on T(reg) levels in lymphoid tissues remain incompletely understood. To explore this issue, we have measured the levels of forkhead box protein 3 (FOXP3)-positive cells and associated immunomodulatory genes in a pathogenic simian immunodeficiency virus/macaque model and found that a loss of T(reg) in lymph nodes occurred following simian immunodeficiency virus infection. Changes in expression of the ligands for CXCR3, CCR4, and CCR7 and the cytokines TGF-beta and IL-2 were all linked to this loss of T(reg), which in turn was linked with increased levels of cellular activation. Our findings identify three mechanisms that likely contribute to SIV-driven loss of T(reg), including reduced levels of cytokines associated with T(reg) differentiation and altered expression of agonist and antagonist chemokines. The loss of T(reg) and the associated cellular activation in lymphoid tissues is consistent with the events in HIV-1-infected individuals and suggest that components of the T(reg) differentiation and trafficking network could be targets for therapeutic intervention. PMID- 18390738 TI - Uropathogenic Escherichia coli block MyD88-dependent and activate MyD88 independent signaling pathways in rat testicular cells. AB - Uropathogenic Escherichia coli (UPEC) is the most common etiological cause of urogenital tract infections and represents a considerable cause of immunological male infertility. We examined TLR 1-11 expression profiles in testicular cells and the functional response to infection with UPEC. All testicular cell types expressed mRNAs for at least two TLRs and, in particular, synthesis of TLR4 was induced in testicular macrophages (TM), Sertoli cells (SC), peritubular cells (PTC), and peritoneal macrophages (PM) after UPEC exposure. Even though MyD88 dependent pathways were activated as exemplified by phosphorylation of mitogen activated protein kinases in TM, SC, PTC, and PM and by the degradation of IkappaBalpha and the nuclear translocation of NF-kappaB in PTC and PM, treatment with UPEC did not result in secretion of the proinflammatory cytokines IL-1alpha, IL-6, and TNF-alpha in any of the investigated cells. Moreover, stimulated production of these cytokines by nonpathogenic commensal E. coli or LPS in PM was completely abolished after coincubation with UPEC. Instead, in SC, PTC, TM, and PM, UPEC exposure resulted in activation of MyD88-independent signaling as documented by nuclear transfer of IFN-related factor-3 and elevated expression of type I IFNs alpha and beta, IFN-gamma-inducible protein 10, MCP-1, and RANTES. We conclude that in this in vitro model UPEC can actively suppress MyD88-dependent signaling at different levels to prevent proinflammatory cytokine secretion by testicular cells. Thus, testicular innate immune defense is shifted to an antiviral-like MyD88-independent response. PMID- 18390740 TI - NK cells play a critical protective role in host defense against acute extracellular Staphylococcus aureus bacterial infection in the lung. AB - Staphylococcus aureus remains a common cause of nosocomial bacterial infections and are often antibiotic resistant. The role of NK cells and IL-15 and their relationship in host defense against extracellular bacterial pathogens including S. aureus remain unclear. We have undertaken several approaches to address this issue using wild type (WT), IL-15 gene knock-out (KO), and NK cell-depleted mouse models. Upon pulmonary staphylococcal infection WT mice had markedly increased activated NK cells, but not NKT or gammadelta T cells, in the airway lumen that correlated with IL-15 production in the airway and with alveolar macrophages. In vitro exposure to staphylococcal products and/or coculture with lung macrophages directly activated NK cells. In contrast, lung macrophages better phagocytosed S. aureus in the presence of NK cells. In sharp contrast to WT controls, IL-15 KO mice deficient in NK cells were found to be highly susceptible to pulmonary staphylococcal infection despite markedly increased neutrophils and macrophages in the lung. In further support of these findings, WT mice depleted of NK cells were similarly susceptible to staphylococcal infection while they remained fully capable of IL-15 production in the lung at levels similar to those of NK competent WT hosts. Our study thus identifies a critical role for NK cells in host defense against pulmonary extracellular bacterial infection and suggests that IL-15 is involved in this process via its indispensable effect on NK cells, but not other innate cells. These findings hold implication for the development of therapeutics in treating antibiotic-resistant S. aureus infection. PMID- 18390739 TI - Utilization of Fc receptors as a mucosal vaccine strategy against an intracellular bacterium, Francisella tularensis. AB - Numerous studies have demonstrated that targeting Ag to Fc receptors (FcR) on APCs can enhance humoral and cellular immunity. However, studies are lacking that examine both the use of FcR-targeting in generating immune protection against infectious agents and the use of FcRs in the induction of mucosal immunity. Francisella tularensis is a category A intracellular mucosal pathogen. Thus, intense efforts are underway to develop a vaccine against this organism. We hypothesized that protection against mucosal infection with F. tularensis would be significantly enhanced by targeting inactivated F. tularensis live vaccine strain (iFt) to FcRs at mucosal sites, via intranasal immunization with mAb-iFt complexes. These studies demonstrate for the first time that: 1) FcR-targeted immunogen enhances immunogen-specific IgA production and protection against subsequent infection in an IgA-dependent manner, 2) FcgammaR and neonatal FcR are crucial to this protection, and 3) inactivated F. tularensis, when targeted to FcRs, enhances protection against the highly virulent SchuS4 strain of F. tularensis, a category A biothreat agent. In summary, these studies show for the first time the use of FcRs as a highly effective vaccination strategy against a highly virulent mucosal intracellular pathogen. PMID- 18390741 TI - Down-modulation of TCR expression by Salmonella enterica serovar Typhimurium. AB - T cell-mediated adaptive immunity is required to help clear infection with the facultative intracellular bacterial pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium), yet development of T cell-mediated adaptive immunity to S. Typhimurium has been described as slow and inefficient. A key step in inducing T cell-mediated adaptive immunity is T cell priming; the activation, proliferation, and differentiation of naive T cells following initial encounter with Ag. We previously demonstrated that S. Typhimurium had a direct inhibitory effect on naive T cells from mouse, blocking their proliferation. In this study, we show that S. Typhimurium down-modulates expression of the TCR beta-chain, a molecule that is essential for Ag recognition and T cell function. Specifically, we demonstrate that reduced amounts of surface and intracellular TCR-beta protein and decreased levels of tcrbeta transcript are expressed by T cells cultured in the presence of S. Typhimurium. We further show that the down-modulation of TCR beta expression requires contact between S. Typhimurium and the T cells and that once contact occurs, a factor capable of reducing TCR-beta expression is secreted. These results provide new insight into the mechanism by which S. Typhimurium may inhibit T cell priming and avoid clearance by the adaptive immune system. PMID- 18390742 TI - Comparison of proteins expressed on secretory vesicle membranes and plasma membranes of human neutrophils. AB - Secretory vesicles are neutrophil intracellular storage granules formed by endocytosis. Understanding the functional consequences of secretory vesicle exocytosis requires knowledge of their membrane proteins. The current study was designed to use proteomic technologies to develop a more complete catalog of secretory vesicle membrane proteins and to compare the proteomes of secretory vesicle and plasma membranes. A total of 1118 proteins were identified, 573 (51%) were present only in plasma membrane-enriched fractions, 418 (37%) only in secretory vesicle-enriched membrane fractions, and 127 (11%) in both fractions. Gene Ontology categorized 373 of these proteins as integral membrane proteins. Proteins typically associated with other intracellular organelles, including nuclei, mitochondria, and ribosomes, were identified in both membrane fractions. Ingenuity Pathway Knowledge Base analysis determined that the majority of canonical and functional pathways were significantly associated with proteins from both plasma membrane-enriched and secretory vesicle-enriched fractions. There were, however, some canonical signaling pathways that involved proteins only from plasma membranes or secretory vesicles. In conclusion, a number of proteins were identified that may elucidate mechanisms and functional consequences of secretory vesicle exocytosis. The small number of common proteins suggests that the hypothesis that secretory vesicles are formed from plasma membranes by endocytosis requires more critical evaluation. PMID- 18390744 TI - HIV envelope suppresses CD4+ T cell activation independent of T regulatory cells. AB - We previously demonstrated that HIV envelope glycoprotein (Env), delivered in the form of a vaccine and expressed by dendritic cells or 293T cells, could suppress Ag-stimulated CD4(+) T cell proliferation. The mechanism remains to be identified but is dependent on CD4 and independent of coreceptor binding. Recently, CD4(+) regulatory T (Treg) cells were found to inhibit protective anti-HIV CD4(+) and CD8(+) T cell responses. However, the role of Tregs in HIV remains highly controversial. HIV Env is a potent immune inhibitory molecule that interacts with host CD4(+) cells, including Treg cells. Using an in vitro model, we investigated whether Treg cells are involved in Env-induced suppression of CD4(+) T cell proliferation, and whether Env directly affects the functional activity of Treg cells. Our data shows that exposure of human CD4(+) T cells to Env neither induced a higher frequency nor a more activated phenotype of Treg cells. Depletion of CD25(+) Treg cells from PBMC did not overcome the Env-induced suppression of CD4(+) T cell proliferation, demonstrating that CD25(+)FoxP3(+) Treg cells are not involved in Env-induced suppression of CD4(+) T cell proliferation. In addition, we extend our observation that similar to Env expressed on cells, Env present on virions also suppresses CD4(+) T cell proliferation. PMID- 18390743 TI - CD127 and CD25 expression defines CD4+ T cell subsets that are differentially depleted during HIV infection. AB - Decreased CD4(+) T cell counts are the best marker of disease progression during HIV infection. However, CD4(+) T cells are heterogeneous in phenotype and function, and it is unknown how preferential depletion of specific CD4(+) T cell subsets influences disease severity. CD4(+) T cells can be classified into three subsets by the expression of receptors for two T cell-tropic cytokines, IL-2 (CD25) and IL-7 (CD127). The CD127(+)CD25(low/-) subset includes IL-2-producing naive and central memory T cells; the CD127(-)CD25(-) subset includes mainly effector T cells expressing perforin and IFN-gamma; and the CD127(low)CD25(high) subset includes FoxP3-expressing regulatory T cells. Herein we investigated how the proportions of these T cell subsets are changed during HIV infection. When compared with healthy controls, HIV-infected patients show a relative increase in CD4(+)CD127(-)CD25(-) T cells that is related to an absolute decline of CD4(+)CD127(+)CD25(low/-) T cells. Interestingly, this expansion of CD4(+)CD127( ) T cells was not observed in naturally SIV-infected sooty mangabeys. The relative expansion of CD4(+)CD127(-)CD25(-) T cells correlated directly with the levels of total CD4(+) T cell depletion and immune activation. CD4(+)CD127( )CD25(-) T cells were not selectively resistant to HIV infection as levels of cell-associated virus were similar in all non-naive CD4(+) T cell subsets. These data indicate that, during HIV infection, specific changes in the fraction of CD4(+) T cells expressing CD25 and/or CD127 are associated with disease progression. Further studies will determine whether monitoring the three subsets of CD4(+) T cells defined based on the expression of CD25 and CD127 should be used in the clinical management of HIV-infected individuals. PMID- 18390745 TI - Protein kinase C-theta critically regulates the proliferation and survival of pathogen-specific T cells in murine listeriosis. AB - Protein kinase C-theta (PKC-theta) is essential for the activation of T cells in autoimmune disorders, but not in viral infections. To study the role of PKC-theta in bacterial infections, PKC-theta(-/-) and wild-type mice were infected with Listeria monocytogenes (LM). In primary and secondary listeriosis, the numbers of LM-specific CD8 and CD4 T cells were drastically reduced in PKC-theta(-/-) mice, resulting in increased CFUs in spleen and liver of both PKC-theta(-/-) C57BL/6 and BALB/c mice. Furthermore, immunization with peptide-loaded wild-type dendritic cells induced LM-specific CD4 and CD8 T cells in wild-type but not in PKC-theta(-/-) mice. In listeriosis, transfer of wild-type T cells into PKC theta(-/-) mice resulted in a normal control of Listeria, and, additionally, a selective expression of PKC-theta in LM-specific T cells was sufficient to drive a normal proliferation and survival of these T cells in LM-infected PKC-theta(-/ ) recipients, illustrating a cell-autonomous function of PKC-theta in LM-specific T cells. Conversely, adoptively transferred PKC-theta(-/-) T cells were partially rescued from cell death and proliferated in LM-infected wild-type recipients, demonstrating that a PKC-theta deficiency of LM-specific T cells can be partially compensated for by a wild-type environment. Additionally, in vitro experiments showed that only the addition of IL-2, but not an inhibition of caspase-3, induced proliferation and prevented death of PKC-theta(-/-) T cells stimulated with LM-infected wild-type dendritic cells, further demonstrating that the impaired proliferation and survival of PKC-theta(-/-) T cells in listeriosis is not intrinsically fixed and can be experimentally improved. PMID- 18390746 TI - Pneumocystis infection enhances antibody-mediated resistance to a subsequent influenza infection. AB - In contrast to the detrimental outcomes most often associated with the resolution of coinfections, the model presented here involving a localized Pneumocystis infection of the lung, followed 2 wk later by an influenza virus infection, results in a significant beneficial outcome for the host. In the week following the influenza infection, immunocompetent coinfected animals exhibited an accelerated rate of virus clearance, an accelerated appearance of higher influenza-specific neutralizing Ab titers in their serum and bronchoalveolar lavage fluid (BALF), significantly reduced inflammatory cytokine levels in their BALF, and reduced levels of morbidity relative to animals infected only with influenza virus. The beneficial outcome observed in coinfected immunocompetent animals was dependent on the ongoing resolution of a viable Pneumocystis infection. No differences in viral clearance were detected between coinfected and influenza-only-infected muMT mice or likewise for SCID mice. The accelerated anti influenza response did not appear to be associated with influenza-specific CD8 T cell-mediated responses or NK cell responses in the lung. Rather, the increased rate of viral clearance was due to the enhancement of the influenza-specific Ab response, which in turn was transiently dependent upon the resolution of the ongoing Pneumocystis infection. PMID- 18390747 TI - Molecular mechanisms of cytokine and chemokine release from eosinophils activated by IL-17A, IL-17F, and IL-23: implication for Th17 lymphocytes-mediated allergic inflammation. AB - IL-17A and IL-17F are members of the IL-17 family that play crucial roles in allergic inflammation. Recent studies reported that IL-17A and IL-17F production from a distinct Th lymphocyte subset, Th17, was specifically induced by IL-23, which was produced by dendritic cells and macrophages in response to microbial stimuli. The IL-23-IL-17 axis might therefore provide a link between infections and allergic diseases. In the present study, we investigated the effects of IL 17A, IL-17F, and IL-23, alone or in combination, on cytokine and chemokine release from eosinophils and the underlying intracellular mechanisms. Human eosinophils were found to constitutively express receptors for IL-17A, IL-17F, and IL-23 at the protein level. IL-17A, IL-17F, and IL-23 could induce the release of chemokines GRO-alpha/CXCL1, IL-8/CXCL8, and MIP-1beta/CCL4 from eosinophils, while IL-17F and IL-23 could also increase the production of proinflammatory cytokines IL-1beta and IL-6. Synergistic effects were observed in the combined treatment of IL-17F and IL-23 on the release of proinflammatory cytokines, and the effects were dose-dependently enhanced by IL-23, but not IL 17F. Further investigations showed that IL-17A, IL-17F, and IL-23 differentially activated the ERK, p38 MAPK, and NF-kappaB pathways. Moreover, inhibition of these pathways using selective inhibitors could significantly abolish the chemokine release induced by IL-17A, IL-17F, and IL-23 and the synergistic increases on IL-1beta and IL-6 production mediated by combined treatment of IL 17F and IL-23. Taken together, our findings provide insight for the Th17 lymphocyte-mediated activation of eosinophils via differential intracellular signaling cascades in allergic inflammation. PMID- 18390748 TI - The tyrosine kinase Pyk2 mediates lipopolysaccharide-induced IL-8 expression in human endothelial cells. AB - Secretion of proinflammatory cytokines by LPS activated endothelial cells contributes substantially to the pathogenesis of sepsis. However, the mechanism involved in this process is not well understood. In the present study, we determined the role of a nonreceptor proline-rich tyrosine kinase, Pyk2, in LPS induced IL-8 (CXCL8) production in endothelial cells. First, we observed a marked activation of Pyk2 in response to LPS. Furthermore, inhibition of Pyk2 activity in these cells by transduction with the catalytically inactive Pyk2 mutant, transfection with Pyk2-specific small interfering RNA, or treatment with Tyrphostin A9 significantly blocked LPS-induced IL-8 production. The supernatants of LPS-stimulated cells exhibiting attenuated Pyk2 activity blocked transendothelial neutrophil migration in comparison to the supernatants of LPS treated controls, thus confirming the inhibition of functional IL-8 production. Investigations into the molecular mechanism of this pathway revealed that LPS activates Pyk2 leading to IL-8 production through the TLR4. In addition, we identified the p38 MAPK pathway to be a critical step downstream of Pyk2 during LPS-induced IL-8 production. Taken together, these results demonstrate a novel role for Pyk2 in LPS-induced IL-8 production in endothelial cells. PMID- 18390749 TI - Kruppel-like factor 4 is essential for inflammatory monocyte differentiation in vivo. AB - Several members of the Kruppel-like factor (KLF) family of transcription factors play important roles in differentiation, survival, and trafficking of blood and immune cell types. We demonstrate in this study that hematopoietic cells from KLF4(-/-) fetal livers (FL) contained normal numbers of functional hematopoietic progenitor cells, were radioprotective, and performed as well as KLF4(+/+) cells in competitive repopulation assays. However, hematopoietic "KLF4(-/-) chimeras" generated by transplantation of KLF4(-/-) fetal livers cells into lethally irradiated wild-type mice completely lacked circulating inflammatory (CD115(+)Gr1(+)) monocytes, and had reduced numbers of resident (CD115(+)Gr1(-)) monocytes. Although the numbers and function of peritoneal macrophages were normal in KLF4(-/-) chimeras, bone marrow monocytic cells from KLF4(-/-) chimeras expressed lower levels of key trafficking molecules and were more apoptotic. Thus, our in vivo loss-of-function studies demonstrate that KLF4, previously shown to mediate proinflammatory signaling in human macrophages in vitro, is essential for differentiation of mouse inflammatory monocytes, and is involved in the differentiation of resident monocytes. In addition, inducible expression of KLF4 in the HL60 human acute myeloid leukemia cell line stimulated monocytic differentiation and enhanced 12-O-tetradecanoylphorbol 13-acetate induced macrophage differentiation, but blocked all-trans-retinoic acid induced granulocytic differentiation of HL60 cells. The inflammation-selective effects of loss-of-KLF4 and the gain-of-KLF4-induced monocytic differentiation in HL60 cells identify KLF4 as a key regulator of monocytic differentiation and a potential target for translational immune modulation. PMID- 18390750 TI - Mechanism of IL-1beta-induced increase in intestinal epithelial tight junction permeability. AB - The IL-1beta-induced increase in intestinal epithelial tight junction (TJ) permeability has been postulated to be an important mechanism contributing to intestinal inflammation of Crohn's disease and other inflammatory conditions of the gut. The intracellular and molecular mechanisms that mediate the IL-1beta induced increase in intestinal TJ permeability remain unclear. The purpose of this study was to elucidate the mechanisms that mediate the IL-1beta-induced increase in intestinal TJ permeability. Specifically, the role of myosin L chain kinase (MLCK) was investigated. IL-1beta caused a progressive increase in MLCK protein expression. The time course of IL-1beta-induced increase in MLCK level correlated linearly with increase in Caco-2 TJ permeability. Inhibition of the IL 1beta-induced increase in MLCK protein expression prevented the increase in Caco 2 TJ permeability. Inhibition of the IL-1beta-induced increase in MLCK activity also prevented the increase in Caco-2 TJ permeability. Additionally, knock-down of MLCK protein expression by small interference RNA prevented the IL-1beta induced increase in Caco-2 TJ permeability. The IL-1beta-induced increase in MLCK protein expression was preceded by an increase in MLCK mRNA expression. The IL 1beta-induced increase in MLCK mRNA transcription and subsequent increase in MLCK protein expression and Caco-2 TJ permeability was mediated by activation of NF kappaB. In conclusion, our data indicate that the IL-1beta increase in Caco-2 TJ permeability was mediated by an increase in MLCK expression and activity. Our findings also indicate that the IL-1beta-induced increase in MLCK protein expression and Caco-2 TJ permeability was mediated by an NF-kappaB-dependent increase in MLCK gene transcription. PMID- 18390752 TI - FcgammaRIIB deficiency leads to autoimmunity and a defective response to apoptosis in Mrl-MpJ mice. AB - Data suggests that modulation of FcgammaRIIB expression represents a significant risk factor for the development of autoimmunity. In this study, we investigated this notion in mice that possess genetics permissible for the development of autoimmunity. To this end, Mrl-MpJ Fcgr2b-/- mice were monitored for the development of autoreactivity. We found that FcgammaRIIB deficiency led to chronic B cell activation associated with increased germinal center and plasma cell accumulation in the spleen. Likewise, Mrl-MpJ Fcgr2b-/- mice exhibited significant serum IgG reactivity against DNA. We further analyzed the IgG isotype contribution to the anti-dsDNA response and found increases in all subtypes with the exception of IgG3. In particular, we found large increases in IgG1 and IgG2b autoreactivity correlating with significant increases in immune complex deposition and kidney pathology. Finally, we found dendritic cells derived from Mrl-MpJ Fcgr2b-/- mice greatly increased IL-12 expression upon coincubation with apoptotic thymocytes compared with wild-type controls. The results indicate that FcgammaRIIB is an important regulator of peripheral tolerance and attenuation of the inhibitory signal it provides enhances autoimmune disease on susceptible backgrounds. Additionally, the data indicates FcgammaRIIB function has a significant impact on APC activity, suggesting a prominent role in dendritic cell activity in response to interaction with particulate autoantigens. PMID- 18390751 TI - A novel proteolytic cascade generates an extracellular matrix-derived chemoattractant in chronic neutrophilic inflammation. AB - Chronic neutrophilic inflammation is a manifestation of a variety of lung diseases including cystic fibrosis (CF). There is increasing evidence that fragments of extracellular matrix proteins, such as collagen and elastin, play an important role in inflammatory cell recruitment to the lung in animal models of airway inflammation. Unfortunately, the association of these peptides with human disease and the identification of therapeutic targets directed toward these inflammatory pathways have remained elusive. In this study, we demonstrate that a novel extracellular matrix-derived neutrophil chemoattractant, proline-glycine proline (PGP), acts through CXC receptors 1 and 2 on neutrophils, similar to N acetylated proline-glycine-proline (N-alpha-PGP). We describe the specific multistep proteolytic pathway involved in PGP generation from collagen, involving matrix metalloproteases 8 and 9 and prolyl endopeptidase, a serine protease for which we identify a novel role in inflammation. PGP generation correlates closely with airway neutrophil counts after administration of proteases in vivo. Using CF as a model, we show that CF sputum has elevated levels of PGP peptides and that PGP levels decline during the course of CF inpatient therapy for acute pulmonary exacerbation, pointing to its role as a novel biomarker for this disease. Finally, we demonstrate that CF secretions are capable of generating PGP from collagen ex vivo and that this generation is significantly attenuated by the use of inhibitors directed toward matrix metalloprotease 8, matrix metalloprotease 9, or prolyl endopeptidase. These experiments highlight unique protease interactions with structural proteins regulating innate immunity and support a role for these peptides as novel biomarkers and therapeutic targets for chronic, neutrophilic lung diseases. PMID- 18390753 TI - CRTH2 plays an essential role in the pathophysiology of Cry j 1-induced pollinosis in mice. AB - PGD(2) is the major prostanoid produced during the acute phase of allergic reactions. Two PGD(2) receptors have been isolated, DP and CRTH2 (chemoattractant receptor-homologous molecule expressed on Th2 cells), but whether they participate in the pathophysiology of allergic diseases remains unclear. We investigated the role of CRTH2 in the initiation of allergic rhinitis in mice. First, we developed a novel murine model of pollinosis, a type of seasonal allergic rhinitis. Additionally, pathophysiological differences in the pollinosis were compared between wild-type and CRTH2 gene-deficient mice. An effect of treatment with ramatroban, a CRTH2/T-prostanoid receptor dual antagonist, was also determined. Repeated intranasal sensitization with Cry j 1, the major allergen of Cryptomeria japonica pollen, in the absence of adjuvants significantly exacerbated nasal hyperresponsive symptoms, Cry j 1-specific IgE and IgG1 production, nasal eosinophilia, and Cry j 1-induced in vitro production of IL-4 and IL-5 by submandibular lymph node cells. Additionally, CRTH2 mRNA in nasal mucosa was significantly elevated in Cry j 1-sensitized mice. Following repeated intranasal sensitization with Cry j 1, CRTH2 gene-deficient mice had significantly weaker Cry j 1-specific IgE/IgG1 production, nasal eosinophilia, and IL-4 production by submandibular lymph node cells than did wild-type mice. Similar results were found in mice treated with ramatroban. These results suggest that the PGD(2)-CRTH2 interaction is elevated following sensitization and plays a proinflammatory role in the pathophysiology of allergic rhinitis, especially pollinosis in mice. PMID- 18390754 TI - Rapid changes in microRNA-146a expression negatively regulate the IL-1beta induced inflammatory response in human lung alveolar epithelial cells. AB - Posttranscriptional regulation of gene expression by microRNAs (miRNAs) has been implicated in the regulation of chronic physiological and pathological responses. In this report, we demonstrate that changes in the expression of miRNAs can also regulate acute inflammatory responses in human lung alveolar epithelial cells. Thus, stimulation with IL-1beta results in a rapid time- and concentration dependent increase in miRNA-146a and, to a lesser extent, miRNA-146b expression, although these increases were only observed at high IL-1beta concentration. Examination of miRNA function by overexpression and inhibition showed that increased miRNA-146a expression negatively regulated the release of the proinflammatory chemokines IL-8 and RANTES. Subsequent examination of the mechanism demonstrated that the action of miRNA-146a was mediated at the translational level and not through the down-regulation of proteins involved in the IL-1beta signaling pathway or chemokine transcription or secretion. Overall, these studies indicate that rapid increase in miRNA-146a expression provides a novel mechanism for the negative regulation of severe inflammation during the innate immune response. PMID- 18390755 TI - Depletion of beta-arrestin-2 promotes tumor growth and angiogenesis in a murine model of lung cancer. AB - Arrestins are adaptor/scaffold proteins that complex with activated and phosphorylated G protein-coupled receptor to terminate G protein activation and signal transduction. These complexes also mediate downstream signaling, independently of G protein activation. We have previously shown that beta arrestin-2 (betaarr2) depletion promotes CXCR2-mediated cellular signaling, including angiogenesis and excisional wound closure. This study was designed to investigate the role of betaarr2 in tumorigenesis using a murine model of lung cancer. To that end, heterotopic murine Lewis lung cancer and tail vein metastasis tumor model systems in betaarr2-deficient mice (betaarr2(-/-)) and control littermates (betaarr2(+/+)) were used. betaarr2(-/-) mice exhibited a significant increase in Lewis lung cancer tumor growth and metastasis relative to betaarr2(+/+) mice. This correlated with decreased number of tumor-infiltrating lymphocytes but with elevated levels of the ELR(+) chemokines (CXCL1/keratinocyte derived chemokine and CXCL2/MIP-2), vascular endothelial growth factor, and microvessel density. NF-kappaB activity was also enhanced in betaarr2(-/-) mice, whereas hypoxia-inducible factor-1alpha expression was decreased. Inhibition of CXCR2 or NF-kappaB reduced tumor growth in both betaarr2(-/-) and betaarr2(+/+) mice. NF-kappaB inhibition also decreased ELR(+) chemokines and vascular endothelial growth factor expression. Altogether, the data suggest that betaarr2 modulates tumorigenesis by regulating inflammation and angiogenesis through activation of CXCR2 and NF-kappaB. PMID- 18390756 TI - Production of type VI collagen by human macrophages: a new dimension in macrophage functional heterogeneity. AB - Macrophages derived from human blood monocytes perform many tasks related to tissue injury and repair. The main effect of macrophages on the extracellular matrix is considered to be destructive in nature, because macrophages secrete metalloproteinases and ingest foreign material as part of the remodeling process that occurs in wound healing and other pathological conditions. However, macrophages also contribute to the extracellular matrix and hence to tissue stabilization both indirectly, by inducing other cells to proliferate and to release matrix components, and directly, by secreting components of the extracellular matrix such as fibronectin and type VIII collagen, as we have recently shown. We now report that monocytes and macrophages express virtually all known collagen and collagen-related mRNAs. Furthermore, macrophages secrete type VI collagen protein abundantly, depending upon their mode of activation, stage of differentiation, and cell density. The primary function of type VI collagen secreted by macrophages appears to be modulation of cell-cell and cell matrix interactions. We suggest that the production of type VI collagen is a marker for a nondestructive, matrix-conserving macrophage phenotype that could profoundly influence physiological and pathophysiological conditions in vivo. PMID- 18390757 TI - Heme oxygenase-1 is a critical regulator of nitric oxide production in enterohemorrhagic Escherichia coli-infected human enterocytes. AB - Enterohemorrhagic Escherichia coli (EHEC) are the causative agent of hemolytic uremic syndrome. In the first stage of the infection, EHEC interact with human enterocytes to modulate the innate immune response. Inducible NO synthase (iNOS) derived NO is a critical mediator of the inflammatory response of the infected intestinal mucosa. We therefore aimed to analyze the role of EHEC on iNOS induction in human epithelial cell lines. In this regard, we show that EHEC down regulate IFN-gamma-induced iNOS mRNA expression and NO production in Hct-8, Caco 2, and T84 cells. This inhibitory effect occurs through the decrease of STAT-1 activation. In parallel, we demonstrate that EHEC stimulate the rapid inducible expression of the gene hmox-1 that encodes for the enzyme heme oxygenase-1 (HO 1). Knock-down of hmox-1 gene expression by small interfering RNA or the blockade of HO-1 activity by zinc protoporphyrin IX abrogated the EHEC-dependent inhibition of STAT-1 activation and iNOS mRNA expression in activated human enterocytes. These results highlight a new strategy elaborated by EHEC to control the host innate immune response. PMID- 18390758 TI - Costimulation of dectin-1 and DC-SIGN triggers the arachidonic acid cascade in human monocyte-derived dendritic cells. AB - Inflammatory mediators derived from arachidonic acid (AA) alter the function of dendritic cells (DC), but data regarding their biosynthesis resulting from stimulation of opsonic and nonopsonic receptors are scarce. To address this issue, the production of eicosanoids by human monocyte-derived DC stimulated via receptors involved in Ag recognition was assessed. Activation of FcgammaR induced AA release, short-term, low-grade PG biosynthesis, and IL-10 production, whereas zymosan, which contains ligands of both the mannose receptor and the human beta glucan receptor dectin-1, induced a wider set of responses including cyclooxygenase 2 induction and biosynthesis of leukotriene C(4) and IL-12p70. The cytosolic phospholipase A(2) inhibitor pyrrolidine 1 completely inhibited AA release stimulated via all receptors, whereas the spleen tyrosine kinase (Syk) inhibitors piceatannol and R406 fully blocked AA release in response to immune complexes, but only partially blocked the effect of zymosan. Furthermore, anti dectin-1 mAb partially inhibited the response to zymosan, and this inhibition was enhanced by mAb against DC-specific ICAM-3-grabbing nonintegrin (SIGN). Immunoprecipitation of DC lysates showed coimmunoprecipitation of DC-SIGN and dectin-1, which was confirmed using Myc-dectin-1 and DC-SIGN constructs in HEK293 cells. These data reveal a robust metabolism of AA in human DC stimulated through both opsonic and nonopsonic receptors. The FcgammaR route depends on the ITAM/Syk/cytosolic phospholipase A(2) axis, whereas the response to zymosan involves the interaction with the C-type lectin receptors dectin-1 and DC-SIGN. These findings help explain the distinct functional properties of DC matured by immune complexes vs those matured by beta-glucans. PMID- 18390759 TI - Evidence for a selective migration of fetus-specific CD4+CD25bright regulatory T cells from the peripheral blood to the decidua in human pregnancy. AB - During pregnancy, the maternal immune system has to tolerate the persistence of fetal alloantigens. Many mechanisms contribute to the prevention of a destructive immune response mediated by maternal alloreactive lymphocytes directed against the allogeneic fetus. Murine studies suggest that CD4(+)CD25(+) T cells provide mechanisms of specific immune tolerance to fetal alloantigens during pregnancy. Previous studies by our group demonstrate that a significantly higher percentage of activated T cells and CD4(+)CD25(bright) T cells are present in decidual tissue in comparison with maternal peripheral blood in human pregnancy. In this study, we examined the phenotypic and functional properties of CD4(+)CD25(bright) T cells derived from maternal peripheral blood and decidual tissue. Depletion of CD4(+)CD25(bright) T cells from maternal peripheral blood demonstrates regulation to third party umbilical cord blood cells comparable to nonpregnant controls, whereas the suppressive capacity to umbilical cord blood cells of her own child is absent. Furthermore, maternal peripheral blood shows a reduced percentage of CD4(+)CD25(bright)FOXP3(+) and CD4(+)CD25(bright)HLA-DR(+) cells compared with peripheral blood of nonpregnant controls. In contrast, decidual lymphocyte isolates contain high percentages of CD4(+)CD25(bright) T cells with a regulatory phenotype that is able to down-regulate fetus-specific and fetus-nonspecific immune responses. These data suggest a preferential recruitment of fetus-specific regulatory T cells from maternal peripheral blood to the fetal-maternal interface, where they may contribute to the local regulation of fetus-specific responses. PMID- 18390760 TI - NK cells expressing a progesterone receptor are susceptible to progesterone induced apoptosis. AB - It has been proposed that progesterone (P4) induces the suppression of immune responses, particularly during pregnancy. However, knowledge about the mechanisms involved has remained largely elusive. We demonstrate herein that peripheral blood NK (PBNK) cells express both classical progesterone receptor (PR) isoforms and are specifically affected by the actions of P4 through two apparently independent mechanisms. Progesterone induces caspase-dependent PBNK cell death, which is reversed by two different anti-progestins, ZK 98.299 and RU 486, supporting the involvement of classical PR isoforms. It was suggested that CD56(bright)CD16(-) killer Ig-like receptor (KIR)(-) NK cells might represent precursor cells, which, upon activation, acquire the features of a more mature NK subset expressing KIR receptors. The present study demonstrates that PR expression seems to be restricted to more mature KIR(+) PBNK cells. The expression of PR had a functional counterpart in the suppressive effect of P4 on IL-12-induced IFN-gamma secretion. This cytokine suppression was mainly observed in KIR(+) PBNK cells, without affecting the high secretion of IFN-gamma by CD56(bright) PBNK cells. The lack of PR expression on CD56(bright)KIR(-) PBNK cells provides an additional phenotypic marker to test the idea that they might represent the PBNK precursors selectively recruited into the endometrium where they differentiate to become the uterine NK cells. Additionally, these findings may be relevant to NK cell function in viral immunity, human reproduction, and tumor immunity. PMID- 18390761 TI - Wikinomics, Steven Blair and the Zurich International Conference on Concussion in Sport. PMID- 18390762 TI - Human Tissue Act: implications for sports science. PMID- 18390768 TI - Effects of long-term training on neutrophil function in male university judoists. AB - OBJECTIVES: To clarify the effects of high-intensity and high-frequency long term/chronic training on neutrophil function and serum levels of myogenic enzymes in male university judoists. METHODS: The subjects were 24 male judoists who had stopped judo training for 6 months and then restarted their training. The following parameters were examined before and after a 2 h unified exercise loading (UEL) at the beginning of the restarted quotidian training (pre-training) and at 2 months, 4 months and 6 months thereafter: myogenic enzymes, neutrophil and leucocyte counts, and neutrophil phagocytic activity (PA) and oxidative burst activity as a measure of reactive oxygen species (ROS) production capability. RESULTS: Myogenic enzymes that were measured after UEL at all four points significantly increased except for creatine kinase at the 2-month point (p<0.01 in each) and neutrophil counts significantly increased after UEL at the pre training, 2-month and 4-month points (p<0.01 in each), but these changes became smaller from the 2-month point. PA significantly decreased after UEL at the pre training and 2-month points (p<0.01 in each), but no change was seen at the 4 month and 6-month points. On the other hand, no change in ROS production per cell after UEL was seen at the pre-training point, but it significantly increased after UEL at the 2-month, 4-month and 6-month points (p<0.01 in each). CONCLUSION: The changing rate of the levels of UEL-mediated myogenic enzymes, neutrophil mobilisation and neutrophil function was seen to decrease at the 2 month, 4-month and 6-month assessments, compared with the pre-training point: these may comprise at least some of the long-term training effects. PMID- 18390769 TI - Gender differences in plantar loading during three soccer-specific tasks. AB - OBJECTIVE: Examine the effect of gender on plantar loading during three football specific tasks. DESIGN: Thirty-four athletes (17 men, 17 women) ran an agility course five times while wearing the Nike Vitoria hard ground cleat. Plantar loading data were recorded during a side cut, a cross-over cut and a forward acceleration task using Pedar-X insoles. SETTING: Controlled laboratory study. PARTICIPANTS: No history of lower extremity injury in the past 6 months, no previous foot or ankle surgery, not currently wearing foot orthotics and play a cleated sport at least two times per week. MAIN OUTCOME MEASUREMENTS: Contact area, maximum force and the force-time integral (FTI) in the medial and lateral midfoot, medial, middle and lateral forefoot as well as the hallux. A univariate ANCOVA (alpha = 0.05) was performed on each dependent variable (covariate was course speed). RESULTS: Significant gender differences existed in the force and force-time integral beneath the lateral midfoot and forefoot during the cross over cut task as well as in the middle forefoot during the side cut task with the men demonstrating an increased force. No significant differences existed in the loading on the medial side of the foot during any tasks. CONCLUSIONS: The results of this study indicate that the increase in plantar loading on the lateral portion of the midfoot and forefoot in men could be one possible explanation for the increased incidence of fifth metatarsal stress fractures in men. Gender differences in loading patterns need to be considered when comparing different movements as well as different footwear conditions. PMID- 18390772 TI - Running economy in early and late maturing youth soccer players does not differ. AB - PURPOSE: The aim of this study was to investigate the influence of maturity on running economy in a population of young soccer players. METHODS: 13 boys (mean age 14.3 years) active in soccer were divided into two groups: 6 early and 7 late maturers. Anthropometrical characteristics, respiratory exchange ratio, heart rate and maximal oxygen uptake were measured. Running economy was assessed at three submaximal running speeds (8, 9.5 and 11 km/h). Allometric coefficients were calculated and used to diminish the effect of body mass. In addition, running style was analysed biomechanically (stride length and meaningful kinematic values). RESULTS: There was no significant difference in the running economy of early and late maturing soccer players, nor any significant differences in mass adjusted physiological values. Therefore physiological differences cannot explain why late maturers succeed in keeping up with early maturers. Late maturing boys take longer relative strides, and have more anteversion of the thigh at heel contact, a smaller knee-angle during swing-phase and a lower mass moment of inertia. CONCLUSION: Running style seems to be an important determinant in running economy of children. PMID- 18390774 TI - Functional assessment of knee joint position sense following anterior cruciate ligament reconstruction. AB - CONTEXT: The anterior cruciate ligament (ACL) has both mechanical and proprioceptive (sensory) functions. Knee-joint proprioception has been assessed using either reproduction of position or threshold to detect passive motion in non-weight-bearing positions. It has been suggested that a more functional and dynamic/active assessment of proprioception might clarify the effects of injury and reconstruction on the proprioceptive function of ACL at the knee joint. OBJECTIVE: To assess knee joint position sense (JPS) following ACL reconstruction in functional positions. SUBJECTS: Twelve male patients who had undergone ACL reconstruction and 12 male healthy control subjects participated in the study. METHODS AND MAIN OUTCOME MEASURES: JPS was evaluated by reproduction of the angles in weight-bearing position with limb movement into flexion and extension. The dominant knee of healthy subjects and both (reconstructed and uninjured) knees of the patients were tested. Absolute angular error was used as a dependent variable. RESULTS: There was no significant difference between the operated and uninjured knees of patients or between patients and healthy controls (p>or=0.05). These results were measured during two tasks of limb movement into flexion and extension. CONCLUSION: We found no evidence of impaired JPS in weight-bearing positions in subjects with ACL reconstruction at a mean follow-up of 11 months after surgery compared with subjects with healthy knees. PMID- 18390777 TI - The stump impingement reflex: a sign of anterior cruciate ligament rupture in the locked knee? PMID- 18390778 TI - Nutrition-focused evaluation and management of dysnatremias. AB - Alterations in sodium and water balance are commonplace and often occur concurrently. Dysnatremias (hyponatremia and hypernatremia) merely reflect a relative excess or deficit of plasma water in relation to plasma sodium. There is no predictable relationship between plasma sodium concentration, extracellular fluid volume, and total body sodium content. Evaluation and management of dysnatremias rely on an understanding of key terminology, normal body fluid composition and distribution, and regulation of water and sodium balance, combined with a directed history and physical and appropriate laboratory tests. Enteral and parenteral nutrition regimens can be modified to help restore normal plasma sodium concentrations. Response to treatment requires close monitoring to avoid potential complications associated with rapid correction of hyponatremia and hypernatremia. PMID- 18390779 TI - Diagnosis and treatment of simple acid-base disorders. AB - The ability to diagnose and treat acid-base disorders is an important component in the practice of the nutrition support clinician. A complete understanding of the basic principles of metabolic and respiratory disorders allows the practitioner to formulate educated decisions regarding fluids, parenteral nutrition salts, and the management of electrolytes. This review will discuss the diagnosis and treatment of common metabolic and respiratory disorders encountered in nutrition support practice. PMID- 18390780 TI - Iron deficiency anemia. AB - The most severe consequence of iron depletion is iron deficiency anemia (IDA), and it is still considered the most common nutrition deficiency worldwide. Although the etiology of IDA is multifaceted, it generally results when the iron demands by the body are not met by iron absorption, regardless of the reason. Individuals with IDA have inadequate intake, impaired absorption or transport, physiologic losses associated with chronological or reproductive age, or chronic blood loss secondary to disease. In adults, IDA can result in a wide variety of adverse outcomes including diminished work or exercise capacity, impaired thermoregulation, immune dysfunction, GI disturbances, and neurocognitive impairment. In addition, IDA concomitant with chronic kidney disease or congestive heart failure can worsen the outcome of both conditions. In this review, the prevalence of IDA related to confounding medical conditions will be described along with its diverse etiologies. Distinguishing IDA from anemia of chronic disease using hematologic measures is reviewed as well. In addition, current diagnostic strategies that are inclusive of clinical presentation, biochemical tests, and differential diagnosis will be outlined, followed by a discussion of treatment modalities and future research recommendations. PMID- 18390781 TI - Magnesium in hypertension, cardiovascular disease, metabolic syndrome, and other conditions: a review. AB - Magnesium plays a role in a number of chronic, disease-related conditions. This article reviews current pertinent literature on magnesium, focusing on hypertension and cardiovascular diseases and implications for relationships with diabetes and metabolic syndrome. A major role for magnesium is in the regulation of blood pressure. While data are not entirely consistent, it does appear that an inverse relationship between magnesium intake and blood pressure is strongest for magnesium obtained from food rather than that obtained via supplements. Hypertension associated with preeclampsia appears to be alleviated when magnesium is administered; in addition, women with adequate intakes of magnesium are less likely to be affected by preeclampsia than those with an inadequate intake. A role for magnesium in other cardiovascular diseases has been noted in that increased magnesium intake may improve serum lipid profiles. Dietary magnesium is also recommended to aid in the prevention of stroke and is important for skeletal growth and development. Magnesium may also play a role in the development of diabetes mellitus, obesity, and metabolic syndrome. There are data from some studies, such as the DASH and PREMIER studies, that suggest that lifestyle changes (including adequate magnesium intake) can benefit blood pressure control, promote weight loss, and improve chronic disease risk. PMID- 18390782 TI - The role of selenium in chronic disease. AB - Selenium functions as a part of proteins known as selenoproteins. Through these selenoproteins, selenium functions as a defensive mechanism for oxidative stress, for the regulation of thyroid hormone activity, and for the redox status of vitamin C and other molecules. In several of its roles, selenium functions as a dietary antioxidant and thus has been studied for its possible role in chronic diseases. This article reviews recent studies regarding selenium status or supplementation in hypertension, cardiovascular disease, cancer, and diabetes mellitus. A few studies regarding aging and mortality are also included. What can be ascertained from this current review is that the maintenance of adequate selenium nutriture and, at minimum, the prevention of a deficiency in selenium would be advisable for all individuals. In addition, the indiscriminant use of selenium supplements should be approached with caution until further randomized, controlled trials monitor the effects of such supplementation, especially on a long-term basis. PMID- 18390783 TI - Correlation of cholestasis with serum copper and whole-blood manganese levels in pediatric patients. AB - Many institutions reduce or eliminate copper (Cu) and manganese (Mn) in parenteral nutrition (PN) solutions when cholestasis develops. Little data exist to support this practice. Fifty-four subjects with known serum Cu, whole-blood Mn, and serum-conjugated bilirubin levels were evaluated in this prospective, observational study. Subjects ranged in weight from 760 g to 65.2 kg. Subjects weighing <25 kg received a daily parenteral dose of 20 microg/kg Cu and 5 microg/kg Mn. Subjects weighing > or =25 kg received a dose of 500 microg/d Cu and 150 microg/d Mn. Cholestasis was defined as a conjugated bilirubin level > or =2 mg/dL. Of the 54 subjects, 20 had cholestasis. Fifteen patients had elevated Cu levels, and 21 had high Mn levels. Seven of the subjects had both high Cu and high Mn levels. The regression model comparing cholestasis as a predictor of high, low, or normal Cu level was not significant (P = .9588). Cholestasis was not a significant predictor of high, low, or normal Mn levels (P = .6533). No correlation between Cu and Mn levels was found. The authors found no significant relationship between conjugated serum bilirubin levels > or =2.0 mg/dL, serum Cu, and whole-blood Mn levels. They found insufficient evidence to support the practice of dosing Mn from a Cu level or vice versa. They recommend obtaining Cu and Mn levels on all pediatric patients who develop cholestasis prior to adjusting parenteral doses and at regular intervals for all long-term PN patients. PMID- 18390784 TI - Death resulting from overzealous total parenteral nutrition: the refeeding syndrome revisited. AB - Commentary is provided on the pivotal paper by Weinsier and Krumdieck from 1981 describing 2 patients who developed profound and fatal refeeding syndrome following initiation of aggressive total parenteral nutrition. This classic description was among the first to describe the overwhelming cardiovascular and pulmonary manifestations that can accompany parenteral refeeding with carbohydrate in chronically malnourished patients. The syndrome has also been described with oral and enteral nutrition. One of the hallmarks of the syndrome is hypophosphatemia. Since 1981, dosing schemes for addressing hypophosphatemia have been refined. Other manifestations of the syndrome include other electrolyte abnormalities such as hypokalemia and hypomagnesemia, hyperglycemia, fluid and sodium retention, and neurologic and hematologic complications. Case reports of refeeding syndrome continue to be published, particularly in the anorexia nervosa population. Stressed, critically ill patients may be at risk of refeeding following short periods of fasting; hypophosphatemia is commonly encountered in this situation. It behooves the current nutrition support practitioner to keep in mind the types of patients at risk of refeeding syndrome and to approach refeeding of such patients with caution and careful monitoring. PMID- 18390785 TI - DPEJ tube placement prevents aspiration pneumonia in high-risk patients. AB - Percutaneous endoscopic gastrostomy (PEG) or PEG tube with transgastric jejunostomy tube (PEG-J) feeding has not been shown to decrease aspiration pneumonia. The aim of this study was to determine if direct percutaneous endoscopic jejunostomy (DPEJ) tube placement results in a decreased incidence of aspiration pneumonia in high-risk patients. The design was a retrospective review of all patients receiving DPEJ tube for aspiration pneumonia from 1999 to 2005. Demographics, incidence of aspiration pneumonia, and outcomes were collected and compared before and after the DPEJ placement. Eleven patients (4 women, 7 men) were identified; their mean age was 44.9 years (range, 18-94 years). The etiologies for recurrent aspiration pneumonia were neurologic disease (9), esophageal surgery (1), and severe debilitation (1). The mean follow-up was 20.9 months (range, 6-48 months). The patients' mean weight increased from 43.8 kg (range, 19-55 kg) to 48.3 kg (range, 30-65 kg) after placement (P < .001). The total number of documented aspiration pneumonia episodes for all patients decreased from 29 (mean, 3.64; range, 1-6) before DPEJ placement to 3 (mean, 0.27; range, 0-2) after DPEJ placement (P < .001). The mean number of aspiration pneumonia events per month prior to the DPEJ placement was 3.39 and postplacement was 0.42 (P < .001). DPEJ placement appears to decrease recurrent aspiration pneumonia in patients with history of aspiration pneumonia. PMID- 18390786 TI - Transnasal endoscopic placement of nasoenteric feeding tubes: outcomes and limitations in non-critically ill patients. AB - Transnasal endoscopic placement of nasoenteric tubes (NETs) has been demonstrated to be useful in the critical care setting, with limited data on its role in non critically ill patients. The authors collected data on consecutive patients from a non-critical care setting undergoing transnasal endoscopic NET placement. All NETs were endoscopically placed using a standard over-the-guidewire technique, and positions were confirmed with fluoroscopy. Patients were monitored until the removal of NETs or death. Twenty-two patients (median age = 62.5 years, 36.4% female) were referred for postpyloric feeding, with main indications of persistent gastrocutaneous fistula (n = 6), gastroparesis or gastric outlet obstruction (n = 5), duodenal stenosis (n = 6), acute pancreatitis (n = 4), and gastroesophageal reflux after surgery (n = 1). Postpyloric placement of NET was achieved in 19 of 22 (86.3%) patients, with 36.8% tube positions in the jejunum, 47.4% in the distal duodenum, and 15.8% in the second part of the duodenum. NET placement was least successful in cases with duodenal stenosis. NETs remained in situ for a median of 24 days (range, 2-94), with tube dislodgement (n = 3) and clogging (n = 5) as the main complications. NET feeding resulted in complete healing of gastrocutaneous fistulae in 5 of 6 patients and provision of total enteral nutrition in 3 of 4 cases of acute pancreatitis and 9 of 11 cases of gastroparesis or proximal duodenal obstruction. Transnasal endoscopy has a role in the placement of NET in non-critically ill patients requiring postpyloric feeding. However, there are some limitations, particularly in cases with altered duodenal anatomy. PMID- 18390787 TI - Review of American Society for Parenteral and Enteral Nutrition (ASPEN) Clinical Guidelines for Nutrition Support in Cancer Patients: nutrition screening and assessment. AB - It is clear that cancer patients develop complex nutrition issues. Nutrition support may or may not be indicated in these patients depending on individual patient characteristics. This review article, the first in a series of articles to examine the A.S.P.E.N. Guidelines for the Use of Parenteral and Enteral Nutrition in Adult and Pediatric Patients Cancer Guidelines, evaluates the evidence related to the use of nutrition screening and nutrition assessment in cancer patients. This first article will provide background concerning nutrition issues in cancer patients as well as discuss the role of nutrition screening and nutrition assessment in the care of cancer patients. The goal of this review is to enrich the discussion contained in the Clinical Guidelines, cite the primary literature more completely, and suggest updates to the guideline statements in light of subsequent published studies. Future articles will explore the guidelines related to nutrition support in oncology patients receiving anticancer therapies. PMID- 18390788 TI - Standards of practice for nutrition support pharmacists. PMID- 18390791 TI - Effect of oral amino acids on counterregulatory responses and cognitive function during insulin-induced hypoglycemia in nondiabetic and type 1 diabetic people. AB - OBJECTIVE: Amino acids stimulate glucagon responses to hypoglycemia and may be utilized by the brain. The aim of this study was to assess the responses to hypoglycemia in nondiabetic and type 1 diabetic subjects after ingestion of an amino acid mixture. RESEARCH DESIGN AND METHODS: Ten nondiabetic and 10 diabetic type 1 subjects were studied on three different occasions during intravenous insulin (2 mU . kg(-1) . min(-1)) plus variable glucose for 160 min. In two studies, clamped hypoglycemia (47 mg/dl plasma glucose for 40 min) was induced and either oral placebo or an amino acid mixture (42 g) was given at 30 min. In the third study, amino acids were given, but euglycemia was maintained. RESULTS: Plasma glucose and insulin were no different in the hypoglycemia studies with both placebo and amino acids (P > 0.2). After the amino acid mixture, plasma amino acid concentrations increased to levels observed after a mixed meal (2.4 +/ 0.13 vs. placebo study 1.7 +/- 0.1 mmol/l, P = 0.02). During clamped euglycemia, ingestion of amino acids resulted in transient increases in glucagon concentrations, which returned to basal by the end of the study. During clamped hypoglycemia, glucagon response was sustained and increased more in amino acid studies versus placebo in nondiabetic and diabetic subjects (P < 0.05), but other counter-regulatory hormones and total symptom score were not different. Beta-OH butyrate was less suppressed after amino acids (200 +/- 15 vs. 93 +/- 9 micromol/l, P = 0.01). Among the cognitive tests administered, the following indicated less deterioration after amino acids than placebo: Trail-Making part B, PASAT (Paced Auditory Serial Addition Test) (2 s), digit span forward, Stroop colored words, and verbal memory tests for nondiabetic subjects; and Trail-Making part B, digit span backward, and Stroop color tests for diabetic subjects. CONCLUSIONS: Oral amino acids improve cognitive function in response to hypoglycemia and enhance the response of glucagon in nondiabetic and diabetic subjects. PMID- 18390792 TI - Novel de novo mutation in sulfonylurea receptor 1 presenting as hyperinsulinism in infancy followed by overt diabetes in early adolescence. AB - OBJECTIVE: Congenital hyperinsulinism, usually associated with severe neonatal hypoglycemia, may progress to diabetes, typically during the 4th decade of life in nonpancreatectomized patients. We aimed to genotype the ATP-sensitive K(+) channel in a 10.5-year-old girl presenting with overt diabetes following hyperinsulinism in infancy. RESEARCH DESIGN AND METHODS: A female aged 10.5 years presented with new-onset, antibody-negative diabetes (A1C 10.6%). She was born large for gestational age (5 kg) to a nondiabetic mother and developed frequent hypoglycemic episodes, which persisted until age 3 years and responded initially to intravenous glucose and later to oral sweets. Currently, she is fully pubertal and obese (BMI 30.2 kg/m(2)), with a partially controlled convulsive disorder (since age 1 year) and poor school performance. Glucose levels were >11.1 mmol/l throughout 72 h of continuous glucose monitoring, with low insulin secretion during intravenous glucose tolerance testing. KCNJ11 and ABCC8 mutation analysis was performed, and the mutation identified was characterized in COSm6 cells. RESULTS: A novel, de novo heterozygous ABCC8 sulfonylurea receptor (SUR)1 mutation (R370S) was identified in the patient's DNA but not in that of either parent. Cotransfection of Kir6.2 and mutant SUR1 demonstrate that the mutated protein is expressed efficiently at the cell surface but fails to respond to MgADP, resulting in minimal channel activity. Interestingly, the heterozygous channel (WT:R370S) responded well to glibenclamide, a finding that lead to the successful initiation of sulfonylurea therapy. CONCLUSIONS: This new ABCC8 mutation is associated with neonatal hyperinsulinism progressing within 10 years to insulinopenic diabetes. Consistent with in vitro findings, the patient responded to sulfonylurea treatment. The mechanism causing the relatively rapid loss in beta-cell function is not clear, but it may involve mutation-induced increased beta-cell apoptosis related to increased metabolic demand. PMID- 18390793 TI - Serum heat shock protein 27 and diabetes complications in the EURODIAB prospective complications study: a novel circulating marker for diabetic neuropathy. AB - OBJECTIVE: Heat shock protein 27 (HSP27) is a member of the small heat shock protein family of proteins. HSP27 expression is enhanced in target tissues of diabetic microvascular complications, and changes in circulating serum HSP27 levels (sHSP27) have been reported in patients with macrovascular disease. We investigated whether sHSP27 levels were associated with micro- and macrovascular complications in type 1 diabetic patients. RESEARCH DESIGN AND METHODS: A cross sectional, nested, case-control study from the EURODIAB Prospective Complications Study of 531 type 1 diabetic patients was performed. Case subjects (n = 363) were defined as those with one or more complications of diabetes; control subjects (n = 168) were defined as those with no evidence of any complication. We measured sHSP27 levels and investigated their associations with diabetes complications. RESULTS: Mean sHSP27 levels were significantly higher in case subjects with distal symmetrical polyneuropathy (DSP) than in control subjects, even after adjustment for age and albumin excretion rate (AER) (785.9 vs. 574.7 pg/ml, P = 0.03). In logistic regression analysis, sHSP27 levels in the upper quartile were associated with a twofold increased odds ratio (OR) of DSP, independently of conventional risk factors, markers of inflammation, and AER (OR 2.41 [95% CI 1.11 5.24]). CONCLUSIONS: In this large cohort of type 1 diabetic subjects, we found an independent association between sHSP27 and DSP. This suggests that sHSP27 levels may be a novel marker for diabetic neuropathy. PMID- 18390794 TI - Voltage-gated ion channels in human pancreatic beta-cells: electrophysiological characterization and role in insulin secretion. AB - OBJECTIVE: To characterize the voltage-gated ion channels in human beta-cells from nondiabetic donors and their role in glucose-stimulated insulin release. RESEARCH DESIGN AND METHODS: Insulin release was measured from intact islets. Whole-cell patch-clamp experiments and measurements of cell capacitance were performed on isolated beta-cells. The ion channel complement was determined by quantitative PCR. RESULTS: Human beta-cells express two types of voltage-gated K(+) currents that flow through delayed rectifying (K(V)2.1/2.2) and large conductance Ca(2+)-activated K(+) (BK) channels. Blockade of BK channels (using iberiotoxin) increased action potential amplitude and enhanced insulin secretion by 70%, whereas inhibition of K(V)2.1/2.2 (with stromatoxin) was without stimulatory effect on electrical activity and secretion. Voltage-gated tetrodotoxin (TTX)-sensitive Na(+) currents (Na(V)1.6/1.7) contribute to the upstroke of action potentials. Inhibition of Na(+) currents with TTX reduced glucose-stimulated (6-20 mmol/l) insulin secretion by 55-70%. Human beta-cells are equipped with L- (Ca(V)1.3), P/Q- (Ca(V)2.1), and T- (Ca(V)3.2), but not N- or R-type Ca(2+) channels. Blockade of L-type channels abolished glucose stimulated insulin release, while inhibition of T- and P/Q-type Ca(2+) channels reduced glucose-induced (6 mmol/l) secretion by 60-70%. Membrane potential recordings suggest that L- and T-type Ca(2+) channels participate in action potential generation. Blockade of P/Q-type Ca(2+) channels suppressed exocytosis (measured as an increase in cell capacitance) by >80%, whereas inhibition of L type Ca(2+) channels only had a minor effect. CONCLUSIONS: Voltage-gated T-type and L-type Ca(2+) channels as well as Na(+) channels participate in glucose stimulated electrical activity and insulin secretion. Ca(2+)-activated BK channels are required for rapid membrane repolarization. Exocytosis of insulin containing granules is principally triggered by Ca(2+) influx through P/Q-type Ca(2+) channels. PMID- 18390795 TI - Tumor necrosis factor-alpha antagonism improves vasodilation during hyperinsulinemia in metabolic syndrome. AB - OBJECTIVE: Obesity is associated with chronic inflammation due to overproduction of proinflammatory cytokines, including tumor necrosis factor (TNF)-alpha. We assessed the effects of TNF-alpha neutralization by infliximab on vascular reactivity during hyperinsulinemia in obesity-related metabolic syndrome. RESEARCH DESIGN AND METHODS: Vascular responses to intra-arterial infusion of acetylcholine (ACh) and sodium nitroprusside (SNP) were assessed in patients with metabolic syndrome, before and after administration of infliximab. RESULTS: Patients had blunted vasodilator responses to ACh and SNP during hyperinsulinemia compared with control subjects; a potentiation of the responsiveness to both ACh and SNP, however, was observed in patients following infliximab. The antioxidant vitamin C improved the vasodilator response to ACh in patients with metabolic syndrome, but its effect was not further enhanced by concurrent administration of infliximab. CONCLUSIONS: TNF-alpha neutralization ameliorates vascular reactivity in metabolic syndrome during hyperinsulinemia, likely in relation to decreased oxidative stress, thereby suggesting an involvement of inflammatory cytokines in vascular dysfunction of these patients. PMID- 18390796 TI - Intake of fruit, vegetables, and fruit juices and risk of diabetes in women. AB - OBJECTIVE: The purpose of this study was to examine the association between fruit, vegetable, and fruit juice intake and development of type 2 diabetes. RESEARCH DESIGN AND METHODS: A total of 71,346 female nurses aged 38-63 years who were free of cardiovascular disease, cancer, and diabetes in 1984 were followed for 18 years, and dietary information was collected using a semiquantitative food frequency questionnaire every 4 years. Diagnosis of diabetes was self-reported. RESULTS: During follow-up, 4,529 cases of diabetes were documented, and the cumulative incidence of diabetes was 7.4%. An increase of three servings/day in total fruit and vegetable consumption was not associated with development of diabetes (multivariate-adjusted hazard ratio 0.99 [95% CI 0.94-1.05]), whereas the same increase in whole fruit consumption was associated with a lower hazard of diabetes (0.82 [0.72-0.94]). An increase of 1 serving/day in green leafy vegetable consumption was associated with a modestly lower hazard of diabetes (0.91 [0.84-0.98]), whereas the same change in fruit juice intake was associated with an increased hazard of diabetes (1.18 [1.10-1.26]). CONCLUSIONS: Consumption of green leafy vegetables and fruit was associated with a lower hazard of diabetes, whereas consumption of fruit juices may be associated with an increased hazard among women. PMID- 18390797 TI - Periodontal disease and incident type 2 diabetes: results from the First National Health and Nutrition Examination Survey and its epidemiologic follow-up study. AB - OBJECTIVE: Type 2 diabetes and periodontal disease are known to be associated, but the temporality of this relationship has not been firmly established. We investigated whether baseline periodontal disease independently predicts incident diabetes over two decades of follow-up. RESEARCH DESIGN AND METHODS: A total of 9,296 nondiabetic male and female National Health and Nutrition Examination Survey (NHANES I) participants aged 25-74 years who completed a baseline dental examination (1971-1976) and had at least one follow-up evaluation (1982-1992) were studied. We defined six categories of baseline periodontal disease using the periodontal index. Of 7,168 dentate participants, 47% had periodontal index = 0 (periodontally healthy); the remaining were classified into periodontal index quintiles. Incident diabetes was defined by 1) death certificate (ICD-9 code 250), 2) self-report of diabetes requiring pharmacological treatment, or 3) health care facility stay with diabetes discharge code. Multivariable logistic regression models assessed incident diabetes odds across increasing levels of periodontal index in comparison with periodontally healthy participants. RESULTS: The adjusted odds ratios (ORs) for incident diabetes in periodontal index categories 1 and 2 were not elevated, whereas the ORs in periodontal index categories 3 through 5 were 2.26 (95% CI 1.56-3.27), 1.71 (1.0-2.69), and 1.50 (0.99-2.27), respectively. The OR in edentulous participants was 1.30 (1.00 1.70). Dentate participants with advanced tooth loss had an OR of 1.70 (P < 0.05) relative to those with minimal tooth loss. CONCLUSIONS: Baseline periodontal disease is an independent predictor of incident diabetes in the nationally representative sample of NHANES I. PMID- 18390798 TI - Dietary behaviors predict glycemic control in youth with type 1 diabetes. AB - OBJECTIVE: To investigate the association between dietary adherence and glycemic control among youth with type 1 diabetes. RESEARCH DESIGN AND METHODS: We conducted a cross-sectional analysis of 119 youth aged 9-14 years (mean +/- SD 12.1 +/- 1.6 years) with diabetes duration > or =1 year (5.4 +/- 3.1 years). Dietary adherence was assessed using the Diabetes Self-Management Profile diet domain. Higher score defined greater dietary adherence. Glycemic control was determined by A1C. RESULTS: Dietary adherence score was inversely correlated with A1C (r = -0.36, P < 0.0001). In a multivariate model (R(2) = 0.34, P < 0.0001), dietary adherence (P = 0.004), pump use (P = 0.03), and caregiver education (P = 0.01) were associated with A1C. A1C of youth in the lowest (9.0%) tertile of diet score was higher than A1C of youth in the middle (8.1%, P = 0.004) and upper (8.4%, P = 0.06) tertiles. Dietary adherence uniquely explained 8% of the variance in A1C in the model. CONCLUSIONS: Greater dietary adherence was associated with lower A1C among youth with type 1 diabetes. PMID- 18390799 TI - Retinol-binding protein 4 and insulin resistance in polycystic ovary syndrome. AB - OBJECTIVE: Polycystic ovary syndrome (PCOS) is an insulin-resistant state with insulin resistance being an established therapeutic target; however, measurement of insulin resistance remains challenging. We aimed to 1) determine serum retinol binding protein 4 (RBP4) levels (purported to reflect insulin resistance) in women with PCOS and control subjects, 2) examine the relationship of RBP4 to conventional markers of insulin resistance, and 3) examine RBP4 changes with interventions modulating insulin resistance in overweight women with PCOS. RESEARCH DESIGN AND METHODS: At baseline, 38 overweight women (BMI >27 kg/m(2)) with PCOS and 17 weight-matched control subjects were compared. Women with PCOS were then randomly assigned to 6 months of a higher-dose oral contraceptive pill (OCP) (35 microg ethinyl estradiol/2 mg cyproterone acetate) or metformin (1 g b.i.d.). Outcome measures were insulin resistance (total insulin area under the curve) on an oral glucose tolerance test, RBP4, and metabolic/inflammatory markers. RESULTS: Overweight women with PCOS were more insulin resistant than control subjects, yet RBP4 levels were not different in women with PCOS versus those in control subjects (35.4 +/- 4.3 vs. 28.9 +/- 3.1 microg/ml, P = 0.36). RBP4 correlated with cholesterol and triglycerides but not with insulin resistance. Metformin improved insulin resistance by 35%, whereas the OCP worsened insulin resistance by 33%. However, RBP4 increased nonsignificantly in both groups (43.7 +/- 6.3 vs. 42.6 +/- 5.5 microg/ml, P = 0.92). CONCLUSIONS: Overweight women with PCOS were more insulin resistant than control subjects, but this finding was not reflected by RBP4 levels. RBP4 correlated with lipid levels but not with insulin resistance markers. RBP4 levels did not change when insulin resistance was reduced by metformin or increased by the OCP. These data suggest that RBP4 is not a useful marker of insulin resistance in PCOS but may reflect other metabolic features of this condition. PMID- 18390800 TI - Association between renal failure and foot ulcer or lower-extremity amputation in patients with diabetes. AB - OBJECTIVE: The objective of this study was to evaluate the association between foot ulcers (DFU) and lower-extremity amputation (LEA) and chronic kidney disease (CKD) in patients with diabetes. RESARCH DESIGN AND METHODS: This was a retrospective cohort study of individuals enrolled between 2002 and 2006 who were aged > or =35 years, had a history of diabetes, and were cared for in general practice. The physicians participated in The Health Information Network of the U.K. RESULTS: The presence of DFU or LEA and estimated glomerular filtration rate (eGFR) were evaluated in 90,617 individuals with a median time of observation of 2.4 years. Of these individuals 378 had an LEA and 2,619 had a DFU. CKD (eGFR <60 ml/min per 1.73 m(2)) was noted in 23,350 (26%) individuals in our cohort. For the development of DFU compared with our reference group (group 1 [eGFR > or =60 ml/min per 1.73 m(2)]), the hazard ratio (HR) for group 2 (eGFR > or =30 and <60 ml/min per 1.73 m(2)) was 1.85 (95% CI 1.71-2.01) and for group 3 (eGFR <30 ml/min per 1.73 m(2)) was 3.92 (3.23-4.75) (all P < 0.001). For LEA, the HR for group 2 was 2.08 (1.68-2.58) and for group 3 was 7.71 (5.29-11.26) (all P < 0.001). CONCLUSIONS: In this observational study, there is a strong association between stage of CKD and DFU or LEA that is probably not just related to the presence of peripheral arterial disease. Individuals with even moderate CKD (eGFR <60 ml/min per 1.73 m(2)) have an increased risk for DFU and LEA. PMID- 18390801 TI - Long-term outcome and disability of diabetic patients hospitalized for diabetic foot ulcers: a 6.5-year follow-up study. AB - OBJECTIVE: The long-term outcome and functional status of subjects hospitalized for diabetic foot ulcers have been poorly studied and thus are the topics of this study. RESEARCH DESIGN AND METHODS: Ninety-four consecutive diabetic subjects hospitalized for diabetic foot ulcers between January 1998 and December 2000 were prospectively followed for mean +/- SD 79.5 +/- 13.3 months. We calculated rates of primary healing, new ulcers, amputations, mortality, and disability and evaluated the global therapeutic success (GTS) of foot care management as defined by the association of primary healing without recurrence or disability at the end of follow-up. RESULTS: Follow-up was successful in 89 of 94 subjects (63 men and 31 women; age 63.7 +/- 10.8 years). Of these, 69 (77.5%) experienced primary healing without major amputation, 39 (43.8%) underwent amputation (24 minor and 15 major), and 46 died (51.7%), including 23 from cardiovascular events. Forty two of 69 patients who experienced primary healing (60.9%) had ulcer recurrence. At the end of the follow-up period, 25 patients (28.1%) were dependent and 40 subjects (44.9%) had achieved GTS. Multivariate analysis showed the role of age as an independent predictor of GTS (P < 0.05) and of impaired renal function/albuminuria as independent predictors of healing failure, first amputation, and mortality (P < 0.01). CONCLUSIONS: Despite a satisfactory initial healing rate, the global long-term outcome of patients hospitalized for diabetic foot ulcers was poor. Nephropathy appears to be an important predictor of long term outcome. Further studies are needed to establish recognized criteria for therapeutic success going beyond just the evaluation of healing rate in the management of diabetic foot ulcers. PMID- 18390802 TI - Plasma carotenoids and onset of dysglycemia in an elderly population: results of the Epidemiology of Vascular Ageing Study. AB - OBJECTIVE: The hypothesis of carotenoid having a preventive role in diabetes is suggested by their antioxidant properties. In this report, we investigated the relationship between baseline total plasma carotenoid levels and 9-year onset of dysglycemia (impaired fasting glucose or type 2 diabetes) in a healthy elderly population. RESEARCH DESIGN AND METHODS: The Epidemiology of Vascular Ageing Study is a 9-year longitudinal study including 1,389 volunteers aged 59-71 years. Fasting plasma glucose was measured at baseline and at 2, 4, and 9 years after inclusion. The relationship between plasma carotenoid at baseline and incidence of dysglycemia was determined by Cox proportional hazards regression analysis adjusting for potential confounders. RESULTS: At 9 years, 127 incident cases of dysglycemia had occurred. Risk of dysglycemia was significantly lower in participants with plasma carotenoid in the highest quartile (Q4) compared with participants in the lowest quartile (Q1) (Q4 vs. Q1: relative risk 0.26 [95% CI 0.14-0.49], P < 10(-4); Q3 vs. Q1: 0.55 [0.34-0.89], P = 0.01; and Q2 vs. Q1: 0.82 [0.51-1.31], P = 0.40). After controlling for sociodemographic variables, lifestyle habits, cardiovascular disease, blood pressure, BMI, and lipid profile, risk of dysglycemia remained significantly lower in participants in the highest quartile of total plasma carotenoid compared with participants in the lowest quartile (Q4 vs. Q1: 0.42 [0.22-0.82], P = 0.01; Q3 vs. Q1: 0.69 [0.41-1.15], P = 0.16; and Q2 vs. Q1: 0.80 [0.48-1.32], P = 0.38). CONCLUSIONS: This study prospectively confirms that plasma carotenoid levels have an independent relationship to onset of dysglycemia. PMID- 18390803 TI - Dietary patterns, insulin resistance, and incidence of type 2 diabetes in the Whitehall II Study. AB - OBJECTIVE: The aim of this study was to identify a dietary pattern associated with insulin resistance and investigate whether this pattern was prospectively associated with type 2 diabetes. RESEARCH DESIGN AND METHODS: Analysis was based on 7,339 participants of the Whitehall II study. Dietary intake was measured using a 127-item food frequency questionnaire. We used the reduced rank regression method to determine dietary patterns using the homeostasis model assessment of insulin resistance as the intermediate or response variable. The association between the dietary pattern identified and incidence of type 2 diabetes was investigated using Cox proportional hazard regression models. RESULTS: We identified a dietary pattern characterized by high consumption of low calorie/diet soft drinks, onions, sugar-sweetened beverages, burgers and sausages, crisps and other snacks, and white bread and low consumption of medium /high-fiber breakfast cereals, jam, French dressing/vinaigrette, and wholemeal bread. Higher dietary pattern scores were associated with increased risk of type 2 diabetes (hazard ratio for top quartile 2.95 [95% CI 2.19-3.97]; adjusted for age, sex, and energy misreporting). This relationship was attenuated after adjustment for ethnicity, employment grade, health behaviors (smoking, alcohol use, and physical activity) but remained significant after further adjustment for blood pressure and BMI (1.51 [1.10-2.09]). CONCLUSIONS: A dietary pattern associated with insulin resistance predicts type 2 diabetes risk after adjustment for a range of confounders. This study adds to the evidence that dietary patterns are an important risk factor for type 2 diabetes. PMID- 18390804 TI - PHYTOCHROME KINASE SUBSTRATE4 modulates phytochrome-mediated control of hypocotyl growth orientation. AB - Gravity and light are major factors shaping plant growth. Light perceived by phytochromes leads to seedling deetiolation, which includes the deviation from vertical hypocotyl growth and promotes hypocotyl phototropism. These light responses enhance survival of young seedlings during their emergence from the soil. The PHYTOCHROME KINASE SUBSTRATE (PKS) family is composed of four members in Arabidopsis (Arabidopsis thaliana): PKS1 to PKS4. Here we show that PKS4 is a negative regulator of both phytochrome A- and B-mediated inhibition of hypocotyl growth and promotion of cotyledon unfolding. Most prominently, pks4 mutants show abnormal phytochrome-modulated hypocotyl growth orientation. In dark-grown seedlings hypocotyls change from the original orientation defined by seed position to the upright orientation defined by gravity and light reduces the magnitude of this shift. In older seedlings with the hypocotyls already oriented by gravity, light promotes the deviation from vertical orientation. Based on the characterization of pks4 mutants we propose that PKS4 inhibits changes in growth orientation under red or far-red light. Our data suggest that in these light conditions PKS4 acts as an inhibitor of asymmetric growth. This hypothesis is supported by the phenotype of PKS4 overexpressers. Together with previous findings, these results indicate that the PKS family plays important functions during light-regulated tropic growth responses. PMID- 18390805 TI - Regulatory network of microRNA399 and PHO2 by systemic signaling. AB - Recently, we showed that microRNA399s (miR399s) control inorganic phosphate (Pi) homeostasis by regulating the expression of PHO2 encoding a ubiquitin-conjugating E2 enzyme 24. Arabidopsis (Arabidopsis thaliana) plants overexpressing miR399 or the pho2 mutant overaccumulate Pi in shoots. The association of Pi translocation and coexpression of miR399s and PHO2 in vascular tissues suggests their involvement in long-distance signaling. In this study, we used reciprocal grafting between wild-type and miR399-overexpressing transgenic plants to dissect the systemic roles of miR399 and PHO2. Arabidopsis rootstocks overexpressing miR399 showed high accumulation of Pi in the wild-type scions because of reduced PHO2 expression in the rootstocks. Although miR399 precursors or expression was not detected, we found a small but substantial amount of mature miR399 in the wild-type rootstocks grafted with transgenic scions, which indicates the movement of miR399 from shoots to roots. Suppression of PHO2 with miR399b or c was less efficient than that with miR399f. Of note, findings in grafted Arabidopsis were also discovered in grafted tobacco (Nicotiana benthamiana) plants. The analysis of the pho1 mutant provides additional support for systemic suppression of PHO2 by the movement of miR399 from Pi-depleted shoots to Pi-sufficient roots. We propose that the long-distance movement of miR399s from shoots to roots is crucial to enhance Pi uptake and translocation during the onset of Pi deficiency. Moreover, PHO2 small interfering RNAs mediated by the cleavage of miR399s may function to refine the suppression of PHO2. The regulation of miR399 and PHO2 via long-distance communication in response to Pi deficiency is discussed. PMID- 18390806 TI - LEUNIG_HOMOLOG and LEUNIG perform partially redundant functions during Arabidopsis embryo and floral development. AB - Transcription corepressors play important roles in animal and plant development. In Arabidopsis (Arabidopsis thaliana), LEUNIG (LUG) and LEUNIG_HOMOLOG (LUH) encode two highly homologous proteins that are similar to the animal and fungal Gro/Tup1-type corepressors. LUG was previously shown to form a putative corepressor complex with another protein, SEUSS (SEU), and to repress the transcription of AGAMOUS in floral organ identity specification. However, the function of LUH is completely unknown. Here, we show that single luh loss-of function mutants develop normal flowers, but lug; luh double mutants are embryo lethal, uncovering a previously unknown function of LUG and LUH in embryonic development. In addition, luh/+ enhances the floral phenotype of lug, revealing a minor role of LUH in flower development. Functional diversification between LUH and LUG is evidenced by the inability of 35S::LUH overexpression to rescue lug mutants and by the opposite expression trends of LUG and LUH in response to biotic and abiotic stresses. The luh-1 mutation does not enhance the defect of seu in flower development, but LUH could directly interact with SEU in yeast. We propose a model that explains the complex relationships among LUH, LUG, and SEU. As most eukaryotes have undergone at least one round of whole-genome duplication during evolution, gene duplication and functional diversification are important issues to consider in uncovering gene function. Our study provides important insights into the complexity in the relationship between two highly homologous paralogous genes. PMID- 18390807 TI - AtOSA1, a member of the Abc1-like family, as a new factor in cadmium and oxidative stress response. AB - The analysis of gene expression in Arabidopsis (Arabidopsis thaliana) using cDNA microarrays and reverse transcription-polymerase chain reaction showed that AtOSA1 (A. thaliana oxidative stress-related Abc1-like protein) transcript levels are influenced by Cd2+ treatment. The comparison of protein sequences revealed that AtOSA1 belongs to the family of Abc1 proteins. Up to now, Abc1-like proteins have been identified in prokaryotes and in the mitochondria of eukaryotes. AtOSA1 is the first member of this family to be localized in the chloroplasts. However, despite sharing homology to the mitochondrial ABC1 of Saccharomyces cerevisiae, AtOSA1 was not able to complement yeast strains deleted in the endogenous ABC1 gene, thereby suggesting different function between AtOSA1 and the yeast ABC1. The atosa1-1 and atosa1-2 T-DNA insertion mutants were more affected than wild type plants by Cd2+ and revealed an increased sensitivity toward oxidative stress (hydrogen peroxide) and high light. The mutants exhibited higher superoxide dismutase activities and differences in the expression of genes involved in the antioxidant pathway. In addition to the conserved Abc1 region in the AtOSA1 protein sequence, putative kinase domains were found. Protein kinase assays in gelo using myelin basic protein as a kinase substrate revealed that chloroplast envelope membrane fractions from the AtOSA1 mutant lacked a 70-kD phosphorylated protein compared to the wild type. Our data suggest that the chloroplast AtOSA1 protein is a new factor playing a role in the balance of oxidative stress. PMID- 18390809 TI - Update on the molecular diagnosis of endocrine tumors: toward -omics-based personalized healthcare? AB - Genetic advances in endocrine neoplasia provided the paradigm for the practice of clinical cancer genetics: germline RET mutations in multiple endocrine neoplasia type 2. In the last 14 yr, both genetics and -omics advances have occurred, almost exponentially in the last 5 yr. The time has come to reevaluate recent advances in genomic medicine's promise to revolutionize personalized healthcare in the context of endocrine neoplasias. This update focuses on two examples of endocrine neoplasias, those of the thyroid and of the adrenal, and discusses recent advances in germline and somatic genetics and genomics, as they relate to clinical application. PMID- 18390808 TI - p38 mitogen-activated protein kinase-dependent tumor necrosis factor-alpha converting enzyme is important for liver injury in hepatotoxic interaction between lipopolysaccharide and ranitidine. AB - Ranitidine (RAN) is one of the drugs associated with idiosyncratic adverse drug reactions (IADRs) in human patients. In rats, cotreatment with nontoxic doses of lipopolysaccharide (LPS) and RAN causes liver injury. This is a potential animal model for RAN-induced IADRs in humans. Previous studies showed that RAN augmented serum tumor necrosis factor (TNF)-alpha production and hepatic neutrophil activation after LPS treatment and that both TNF-alpha and neutrophils are crucial for the liver pathogenesis. We tested the hypothesis that p38 mitogen activated protein kinase activation is necessary for TNF-alpha production, neutrophil activation, and subsequent liver injury. LPS/RAN cotreatment caused more p38 activation compared with LPS alone. The p38 inhibitor SB 239063 [trans-1 (4-hydroxycyclohexyl)-4-(4-fluorophenyl)-5-(2-methoxypyridimidin-4-yl) imidazole] reduced liver injury in rats cotreated with LPS/RAN. This inhibitor also reduced neutrophil activation and attenuated hemostatic system activation. SB 239063 decreased serum TNF-alpha concentration after LPS/RAN treatment to the same level as LPS treatment. However, the inhibitor did not reduce TNF-alpha mRNA in liver, suggesting a post-transcriptional mode of action. This might occur through TNF alpha-converting enzyme (TACE), which cleaves pro-TNF-alpha into its active form. Indeed, a TACE inhibitor administered just before RAN treatment reduced serum TNF alpha protein. The TACE inhibitor also reduced liver injury and serum plasminogen activator inhibitor (PAI)-1. Furthermore, a PAI-1 inhibitor reduced neutrophil activation and liver injury after LPS/RAN treatment. In summary, RAN enhanced TNF alpha production after LPS treatment through augmented p38 activation, and this seems to occur through TACE. The prolonged TNF-alpha production enhanced PAI-1 production after RAN cotreatment, and this is important for the hepatotoxicity. PMID- 18390810 TI - Massive soft tissue calcifications and cinacalcet. PMID- 18390811 TI - Thyrotropin suppressive therapy in thyroid carcinoma: what are the targets? PMID- 18390812 TI - Do cardiovascular risk factors in polycystic ovarian syndrome result in more cardiovascular events? PMID- 18390813 TI - Thyroid hormone treatment to mend a broken heart. PMID- 18390814 TI - Reproductive hormones and skeletal health in young women. PMID- 18390815 TI - Whole-body dual-energy x-ray absorptiometry comes of age: bone structural measures and their physiological determinants in anorexia nervosa. PMID- 18390816 TI - Should we still use glucose-suppressed growth hormone levels for the evaluation of acromegaly? PMID- 18390817 TI - Caveolin-1: a new locus for human lipodystrophy. PMID- 18390818 TI - Test-retest reliability of the olfactory detection threshold test of the Sniffin' sticks. AB - The aim of the present study was to investigate the test-retest reliability of the olfactory detection threshold subtest of the Sniffin' Sticks test battery, if administered repeatedly on 4 time points. The detection threshold test was repeatedly conducted in 64 healthy subjects. On the first testing session, the threshold test was accomplished 3 times (T(1) = 0 min, T(2) = 35 min, and T(3) = 105 min), representing a short-term testing. A fourth threshold test was conducted on a second testing session (T(4) = 35.1 days after the first testing session), representing a long-term testing. The average scores for olfactory detection threshold for n-butanol did not differ significantly across the 4 points of time. The test-retest reliability (Pearson's r) between the 4 time points of threshold testing were in a range of 0.43-0.85 (P < 0.01). These results support the notion that the olfactory detection threshold test is a highly reliable method for repeated olfactory testing, even if the test is repeated more than once per day and over a long-term period. It is concluded that the olfactory detection threshold test of the Sniffin' Sticks is suitable for repeated testing during experimental or clinical studies. PMID- 18390819 TI - Angiotensin II type 1 receptor blocker preserves tolerance to ischemia reperfusion injury in Dahl salt-sensitive rat heart. AB - Oxidative stress is involved in the tolerance to ischemia-reperfusion (I/R) injury. Because angiotensin II type 1 receptor blockers (ARBs) inhibit oxidative stress, there is concern that ARBs abolish the tolerance to I/R injury. Dahl salt sensitive (DS) hypertensive and salt-resistant (DR) normotensive rats received an antioxidant, 2-mercaptopropionylglycine (MPG), or an ARB, losartan, for 7 days. Losartan and MPG significantly inhibited oxidative stress as determined by tissue malondialdehyde + 4-hydroxynoneal and increased expression of inducible nitric oxide synthase (iNOS) in the DS rat heart. However, losartan but not MPG activated endothelial nitric oxide synthase (eNOS) as assessed by phosphorylation of eNOS on Ser1177. Infarct size after 30-min left coronary artery occlusion followed by 2-h reperfusion was comparable between DS and DR rat hearts. Although MPG and losartan had no effect on infarct size in the DR rat heart, MPG but not losartan significantly increased infarct size in the DS rat heart. A selective iNOS inhibitor, 1400W, increased infarct size in the DS rat heart, but it had no effect on infarct size in the losartan-treated DS rat heart. However, a nonselective NOS inhibitor, Nomega-nitro-l-arginine methyl ester, increased infarct size in the losartan-treated DS rat heart. These results suggest that losartan preserves the tolerance to I/R injury by activating eNOS despite elimination of redox-sensitive upregulation of iNOS and iNOS-dependent cardioprotection in the DS rat heart. PMID- 18390820 TI - Mouse heart valve structure and function: echocardiographic and morphometric analyses from the fetus through the aged adult. AB - The purpose of this study is to provide standard echocardiographic and morphometric data for normal mouse valve structure and function from late fetal to aged adult stages. Cross-sectional, two-dimensional and Doppler transthoracic echocardiography was performed in C57BL6 mice anesthetized with 1% to 2% isoflurane at embryonic day 18.5 (late fetal), 10 days (neonate), 1 mo (juvenile), 2 mo (young adult), 9 mo (old adult), and 16 mo (aged adult). Normal annulus dimensions indexed to age or weight, and selected flow velocities, were established by echocardiography. After echocardiographic imaging, hearts were harvested and histological and morphometric analyses were performed. Morphometric analysis demonstrated a progressive valve thinning and elongation during the fetal and juvenile stages that plateaued during adult stages (ANOVA, P < 0.01); however, there was increased thickening of the hinge of the aortic valve with advanced age, reminiscent of human aortic valve sclerosis. There was no age related calcification. The results of this study provide comprehensive echocardiographic and morphometric data for normal mouse valve structure and function from late fetal to aged adult stages and should prove useful as a reference standard for future studies using mouse models of progressive valve disease. PMID- 18390821 TI - Impaired capsaicin-induced relaxation of coronary arteries in a porcine model of the metabolic syndrome. AB - Recent studies implicate channels of the transient receptor potential vanilloid family (e.g., TRPV1) in regulating vascular tone; however, little is known about these channels in the coronary circulation. Furthermore, it is unclear whether metabolic syndrome alters the function and/or expression of TRPV1. We tested the hypothesis that TRPV1 mediates coronary vasodilation through endothelium dependent mechanisms that are impaired by the metabolic syndrome. Studies were conducted on coronary arteries from lean and obese male Ossabaw miniature swine. In lean pigs, capsaicin, a TRPV1 agonist, relaxed arteries in a dose-dependent manner (EC50 = 116 +/- 41 nM). Capsaicin-induced relaxation was blocked by the TRPV1 antagonist capsazepine, endothelial denudation, inhibition of nitric oxide synthase, and K+ channel antagonists. Capsaicin-induced relaxation was impaired in rings from pigs with metabolic syndrome (91 +/- 4% vs. 51 +/- 10% relaxation at 100 microM). TRPV1 immunoreactivity was prominent in coronary endothelial cells. TRPV1 protein expression was decreased 40 +/- 11% in obese pigs. Capsaicin (100 microM) elicited divalent cation influx that was abolished in endothelial cells from obese pigs. These data indicate that TRPV1 channels are functionally expressed in the coronary circulation and mediate endothelium-dependent vasodilation through a mechanism involving nitric oxide and K+ channels. Impaired capsaicin-induced vasodilation in the metabolic syndrome is associated with decreased expression of TRPV1 and cation influx. PMID- 18390822 TI - Mast cell stabilization improves cardiac contractile function following hemorrhagic shock and resuscitation. AB - Hemorrhagic shock (HS) is associated with cardiac contractile dysfunction. Mast cell (MC) degranulation is hypothesized to mediate the cardiodepressant effect. Cardiac function was assessed after HS and resuscitation (HS/R) with the administration of the MC stabilizers to prevent MC degranulation. Anesthetized male Sprague-Dawley rats were randomized to sham-operated control or HS/R groups and underwent 60 min of HS followed by 2 h of resuscitated reperfusion. Animals in the HS/R groups were randomized to receive cromolyn (5 mg/kg), ketotifen (1 mg/kg), or saline 15 min before shock. Hearts were excised following HS or 2 h of reperfusion, and function was assessed on a Langendorff apparatus. A second group of randomized animals had serial blood samples taken to assess MC degranulation by quantifying levels of serum beta-hexosaminidase. Hearts were excised at 0 min (before HS) and following 60 min of HS (before resuscitation) for a histological evaluation of MC density and degranulation. In vivo MC stabilization using ketotifen and cromolyn improved cardiac peak systolic pressure (P < 0.05), contractility (P < 0.05), and relaxation (P < 0.05) compared with that of HS controls. Serum beta-hexosaminidase increased during HS/R and was inhibited by MC stabilization (P < 0.05). Degranulation was inhibited when assessed by histochemistry and immune fluorescence. The inhibition of MC degranulation can significantly improve cardiac function following HS/R. PMID- 18390823 TI - Surfactant protein D: not just for the lung anymore. PMID- 18390824 TI - The workings of a multicultural research team. AB - PURPOSE: Transcultural nurse researchers are exposed to the challenges of developing and maintaining a multiethnic team. With the example of a multicultural research study of family caregivers conducted in the Miami-Dade area, the authors guide the readers through steps of developing a culturally competent and effective team. DESIGN: Pointing out challenges and successes, the authors illustrate team processes and successful strategies relative to recruitment of qualified members, training and team maintenance, and evaluation of team effectiveness. METHOD: With relevant concepts from the literature applied to practical examples, the authors demonstrate how cultural team competence grows in a supportive work environment. PMID- 18390825 TI - Certified nursing assistants' explanatory models of nursing home resident depression. AB - In this study, the authors describe certified nursing assistants' (CNA) Explanatory Models (EMs) of depression and aspects of their EMs that may contribute to the underdetection of depression in nursing homes. Interviews with 18 CNAs working in two nursing homes are guided by Kleinman's Explanatory Models of Illness framework. Interview data are content analyzed and CNAs' descriptions of depression are compared to the MDS 2.0 Mood Screening criteria and to the DSM IV-TR diagnostic criteria for depression. The result is that the CNAs are unsure about the duration and normalcy of depression in residents. Although they have no formal training, CNAs feel responsible for observing for signs of depression and describe verbal and nonverbal ways of interacting when providing emotional care to residents. CNAs hold potential to improve the detection of depression and contribute to the emotional care of residents. Attention to staff knowledge deficits and facility barriers may enhance this potential. PMID- 18390827 TI - GSK-3/beta-catenin signaling axis in airway smooth muscle: role in mitogenic signaling. AB - Beta-catenin plays a dual role in cellular signaling by stabilizing cadherin mediated cell-cell contact and by regulating gene transcription associated with cell cycle progression. Nonetheless, its presence and function in airway smooth muscle have not been determined. We hypothesized a central role for beta-catenin in mitogenic signaling in airway smooth muscle in response to growth factor stimulation. Immunocytochemical and biochemical analysis revealed that human airway smooth muscle cells indeed express abundant beta-catenin, which was localized primarily to the plasma membrane in quiescent cells. Treatment of airway smooth muscle cells with PDGF or FBS induced sustained phosphorylation of glycogen synthase kinase-3 (GSK-3), a negative regulator in its unphosphorylated form that promotes beta-catenin degradation. GSK-3 phosphorylation was also increased in airway smooth muscle cells with a proliferative phenotype compared with quiescent airway smooth muscle cells with a mature phenotype. Parallel with the increase in GSK-3 phosphorylation, growth factor treatment induced an increased expression and nuclear presence of beta-catenin and activated promitogenic signaling in airway smooth muscle, including the phosphorylation of retinoblastoma protein, DNA synthesis ([(3)H]thymidine incorporation), and cell proliferation. Importantly, small interfering RNA knockdown of beta-catenin strongly reduced retinoblastoma protein phosphorylation, [(3)H]thymidine incorporation, and cell proliferation induced by PDGF and FBS. Collectively, these data reveal the existence of a GSK-3/beta-catenin signaling axis in airway smooth muscle that is regulated by growth factors and of central importance to mitogenic signaling. PMID- 18390828 TI - MMP-12 induces IL-8/CXCL8 secretion through EGFR and ERK1/2 activation in epithelial cells. AB - Macrophage metalloelastase (MMP-12) is described to be involved in pulmonary inflammatory response. To determine the mechanisms linking MMP-12 and inflammation, we examined the effect of recombinant human MMP-12 (rhMMP-12) catalytic domain on IL-8/CXCL8 production in cultured human airway epithelial (A549) cells. Stimulation with rhMMP-12 resulted in a concentration-dependent IL 8/CXCL8 synthesis 6 h later. Similar results were also observed in cultured BEAS 2B bronchial epithelial cells. In A549 cells, synthetic matrix metalloproteinase (MMP) inhibitors prevented rhMMP-12-induced IL-8/CXCL8 release. We further demonstrated that in A549 cells, rhMMP-12 induced transient, peaking at 5 min, activation of ERK1/2. Selective MEK inhibitors (U0126 and PD-98059) blocked both IL-8/CXCL8 release and ERK1/2 phosphorylation. IL-8/CXCL8 induction and ERK1/2 activation were preceded by EGF receptor (EGFR) tyrosine phosphorylation, within 2 min, and reduced by selective EGFR tyrosine kinase inhibitors (AG-1478 and PD168393) by a neutralizing EGFR antibody and by small interfering RNA oligonucleotides directed against EGFR, implicating EGFR activation. In addition, we observed an activation of c-Fos in A549 cells stimulated by rhMMP-12, dependent on ERK1/2. Using small interfering technique, we showed that c-Fos is involved in rhMMP-12-induced IL-8/CXCL8 production. From these results, we conclude that one mechanism, by which MMP-12 induces IL-8/CXCL8 release from the alveolar epithelium, is the EGFR/ERK1/2/activating protein-1 pathway. PMID- 18390826 TI - Structural glycobiology: a game of snakes and ladders. AB - Oligo- and polysaccharides are infamous for being extremely flexible molecules, populating a series of well-defined rotational isomeric states under physiological conditions. Characterization of this heterogeneous conformational ensemble has been a major obstacle impeding high-resolution structure determination of carbohydrates and acting as a bottleneck in the effort to understand the relationship between the carbohydrate structure and function. This challenge has compelled the field to develop and apply theoretical and experimental methods that can explore conformational ensembles by both capturing and deconvoluting the structural and dynamic properties of carbohydrates. This review focuses on computational approaches that have been successfully used in combination with experiment to detail the three-dimensional structure of carbohydrates in a solution and in a complex with proteins. In addition, emerging experimental techniques for three-dimensional structural characterization of carbohydrate-protein complexes and future challenges in the field of structural glycobiology are discussed. The review is divided into five sections: (1) The complexity and plasticity of carbohydrates, (2) Predicting carbohydrate-protein interactions, (3) Calculating relative and absolute binding free energies for carbohydrate-protein complexes, (4) Emerging and evolving techniques for experimental characterization of carbohydrate-protein structures, and (5) Current challenges in structural glycoscience. PMID- 18390829 TI - Expression of the proapoptotic protein Bax is reduced in bronchial mucous cells of asthmatic subjects. AB - The present studies were designed to determine whether our findings in mice showing that the Bcl-2-associated protein X (Bax), which plays a role in the resolution of allergen-induced mucous cell metaplasia, can be applied to asthma in humans. Immunostaining of autopsy tissues from mild and severe asthmatic subjects showed a significant reduction in the percentage of Bax-positive mucous cells compared with those from nonasthmatic controls. To exclude the possibility that postmortem changes may have affected Bax expression, Bax mRNA levels in airway epithelial cells obtained from nonsmoking asthmatic subjects were compared with those from nonasthmatic controls. Because the number of cells obtained by bronchial brushings is limited, we developed a robust preamplification procedure of cDNA before quantitative real-time PCR to allow detection of 100 gene targets from limited sample size, even when it was prepared from partially degraded RNA. cDNA was prepared by reverse transcription from RNA isolated from bronchial epithelial cells obtained by bronchial brushings from well-characterized subjects without lung disease and from subjects with mild asthma. Quantitative analysis showed that Bax mRNA levels were significantly reduced in samples obtained from asthma patients compared with nonasthma controls. Furthermore, Bax mRNA levels were reduced when primary airway epithelial cells from 10 individuals were treated in culture with the T helper 2 cytokine IL-13. These studies show that Bax expression is reduced in airway epithelial cells of even mild asthmatic subjects and suggest that restoring Bax expression may provide a clinical approach for restoring the normal numbers of epithelial cells and reduced mucous hypersecretion in asthma. PMID- 18390830 TI - Endoplasmic reticulum stress in alveolar epithelial cells is prominent in IPF: association with altered surfactant protein processing and herpesvirus infection. AB - Recent evidence suggests that dysfunctional type II alveolar epithelial cells (AECs) contribute to the pathogenesis of idiopathic pulmonary fibrosis (IPF). Based on the hypothesis that disease-causing mutations in surfactant protein C (SFTPC) provide an important paradigm for studying IPF, we investigated a potential mechanism of AEC dysfunction suggested to result from mutant SFTPC expression: induction of endoplasmic reticulum (ER) stress and the unfolded protein response (UPR). We evaluated biopsies from 23 IPF patients (including 3 family members with L188Q SFTPC mutations, 10 individuals with familial interstitial pneumonia without SFTPC mutations, and 10 individuals with sporadic IPF) and sections from 10 control lungs. After demonstrating UPR activation in cultured A549 cells expressing mutant SFTPC, we identified prominent expression of UPR markers in AECs in the lungs of patients with SFTPC mutation-associated fibrosis. In individuals with familial interstitial pneumonia without SFTPC mutations and patients with sporadic IPF, we also found UPR activation selectively in AECs lining areas of fibrotic remodeling. Because herpesviruses are found frequently in IPF lungs and can induce ER stress, we investigated expression of viral proteins in lung biopsies. Herpesvirus protein expression was found in AECs from 15/23 IPF patients and colocalized with UPR markers in AECs from these patients. ER stress and UPR activation are found in the alveolar epithelium in patients with IPF and could contribute to disease progression. Activation of these pathways may result from altered surfactant protein processing or chronic herpesvirus infection. PMID- 18390831 TI - RAGE: developmental expression and positive feedback regulation by Egr-1 during cigarette smoke exposure in pulmonary epithelial cells. AB - The receptor for advanced glycation end-products (RAGE) is a member of the immunoglobin superfamily of multiligand receptors. Following ligand binding, mechanisms associated with host defense, tissue remodeling, and inflammation are activated. RAGE is highly expressed in pulmonary epithelium transitioning from alveolar type (AT) II to ATI cells and is upregulated in the presence of ligand; however, the regulation and function of RAGE during development are less clear. Herein, immunohistochemistry demonstrated a temporal-spatial pattern of RAGE expression in pulmonary epithelial cells from embryonic day 17.5 to postnatal day 10. Cotransfection experiments revealed that the mouse RAGE promoter was activated by early growth response gene 1 (Egr-1) and inhibited by thyroid transcription factor-1 (TTF-1) via interaction with specific regulatory elements. A rat ATI cell line (R3/1) with endogenous RAGE expression also differentially regulated RAGE when transfected with TTF-1 or Egr-1. Because Egr-1 is markedly induced in pulmonary epithelial cells exposed to cigarette smoke extract (CSE; Reynolds PR, Hoidal JR. Am J Respir Cell Mol Biol 35: 314-319, 2006.), we sought to investigate RAGE induction by CSE. Employing RT-PCR and Western blotting, RAGE and common ligands (amphoterin and S100A12) were upregulated in epithelial (R3/1 and A549) and macrophage (RAW) cell lines following exposure to CSE. Immunostaining for RAGE in cells similarly exposed and in lungs from mice exposed to cigarette smoke for 6 mo revealed elevated RAGE expression in pulmonary epithelium. After the addition of glyoxylated BSA, an advanced glycation end product that binds RAGE, real-time RT-PCR detected a 200-fold increase in Egr-1. These results indicate that Egr-1 regulates RAGE expression during development and the likelihood of positive feedback involving Egr-1 and RAGE in cigarette smoke-related disease. PMID- 18390832 TI - Impact of the FcgammaII-receptor on quartz uptake and inflammatory response by alveolar macrophages. AB - The inflammatory response following particle inhalation is described as a key event in the development of lung diseases, e.g., fibrosis and cancer. The essential role of alveolar macrophages (AM) in the pathogenicity of particles through their functions in lung clearance and mediation of inflammation is well known. However, the molecular mechanisms and direct consequences of particle uptake are still unclear. Inhibition of different classic phagocytosis receptors by flow cytometry shows a reduction of the dose-dependent quartz particle (DQ12) uptake in the rat AM cell line NR8383. Thereby the strongest inhibitory effect was observed by blocking the FcgammaII-receptor (FcgammaII-R). Fluorescence immunocytochemistry, demonstrating FcgammaII-R clustering at particle binding sites as well as transmission electron microscopy, visualizing zippering mechanism-like morphological changes, confirmed the role of the FcgammaII-R in DQ12 phagocytosis. FcgammaII-R participation in DQ12 uptake was further strengthened by the quartz-induced activation of the Src-kinase Lyn, the phospho tyrosine kinases Syk (spleen tyrosine kinase) and PI3K (phosphatidylinositol 3 kinase), as shown by Western blotting. Activation of the small GTPases Rac1 and Cdc42, shown by immunoprecipitation, as well as inhibition of tyrosine kinases, GTPases, or Rac1 provided further support for the role of the FcgammaII-R. Consistent with the uptake results, FcgammaII-R activation with its specific ligand caused a similar generation of reactive oxygen species and TNF-alpha release as observed after treatment with DQ12. In conclusion, our results indicate a major role of FcgammaII-R and its downstream signaling cascade in the phagocytosis of quartz particles in AM as well as in the associated generation and release of inflammatory mediators. PMID- 18390833 TI - Pyrrolidine dithiocarbamate restores endothelial cell membrane integrity and attenuates monocrotaline-induced pulmonary artery hypertension. AB - Monocrotaline (MCT)-induced pulmonary artery hypertension (PAH) in rats is preceded by an inflammatory response, progressive endothelial cell membrane disruption, reduction in the expression of caveolin-1, and reciprocal activation of STAT3 (PY-STAT3). Superoxide and NF-kappaB have been implicated in PAH. To evaluate the role of caveolin-1, PY-STAT3 activation, and superoxide in PAH, MCT injected rats were treated daily with pyrrolidine dithiocarbamate (PDTC; starting on days 1, 3, and 14 x 2 wk), an inhibitor of inflammation and NF-kappaB activation. Hemodynamic data, the expression of inhibitory (I)-kappaBalpha, caveolin-1, and Tie2 (a membrane protein), activation of PY-STAT3 and NF-kappaB, and superoxide chemiluminescence were examined. Rats developed progressive PAH at 2 wk post-MCT. There was progressive reduction in the expression of caveolin-1, Tie2, and activation of PY-STAT3 in the lungs. Reduction in I-kappaBalpha expression was present at 2 and 4 wk post-MCT. Superoxide chemiluminescence and NF-kappaB activation were observed only at 2 wk post-MCT and both decreased by 4 wk post-MCT despite progressive PAH. PDTC (starting on days 1 and 3) rescued caveolin-1 and Tie2, reversed MCT-induced PY-STAT3 activation, and attenuated PAH. In addition, PDTC restored I-kappaBalpha expression and reduced superoxide chemiluminescence at 2 wk but did not inhibit NF-kappaB activation despite attenuation of PAH. PDTC had no effect on established PAH. Increased superoxide chemiluminescence and NF-kappaB activation appear to be a transient phenomenon in the MCT model. Thus the disruption of endothelial cell membrane integrity resulting in caveolin-1 loss and reciprocal activation of PY-STAT3 plays a key role in the MCT-induced PAH. PMID- 18390834 TI - The human zinc transporter SLC39A8 (Zip8) is critical in zinc-mediated cytoprotection in lung epithelia. AB - Zinc is an essential micronutrient and cytoprotectant involved in the host response to inflammatory stress. We tested whether zinc transporters, the critical regulators that maintain intracellular zinc concentrations, play a role in cell survival, particularly in lung epithelia, during inflammation. Initially, mRNA transcripts were quantitatively measured by RT-PCR for all known human zinc transporters, including 14 importers (SLC39A(1-14)) and 10 exporters (SLC30A(1 10)), in primary human lung epithelia obtained from multiple human donors and BEAS-2B cell cultures under baseline and TNF-alpha-stimulated conditions. While many zinc transporters were constitutively expressed, only SLC39A8 (Zip8) mRNA was strongly induced by TNF-alpha. Endogenous Zip8 protein was not routinely detected under baseline conditions. In sharp contrast, TNF-alpha induced the expression of a glycosylated protein that translocated to the plasma membrane and mitochondria. Increased Zip8 expression resulted in an increase in intracellular zinc content and coincided with cell survival in the presence of TNF-alpha. Inhibition of Zip8 expression using a short interfering RNA probe reduced cellular zinc content and impaired mitochondrial function in response to TNF alpha, resulting in loss of cell viability. These data are the first to characterize human Zip8 and remarkably demonstrate that upregulation of Zip8 is sufficient to protect lung epithelia against TNF-alpha-induced cytotoxicity. We conclude that Zip8 is unique, relative to other Zip proteins, by functioning as an essential zinc importer at the onset of inflammation, thereby facilitating cytoprotection within the lung. PMID- 18390835 TI - PPARbeta/delta agonist stimulates human lung carcinoma cell growth through inhibition of PTEN expression: the involvement of PI3K and NF-kappaB signals. AB - Recent studies suggest that activation of peroxisome proliferator-activated receptor beta/delta (PPARbeta/delta) promotes cancer cell survival. We previously demonstrated that a selective PPARbeta/delta agonist, GW501516, stimulated human non-small cell lung carcinoma (NSCLC) cell growth. Here, we explore the mechanisms responsible for this effect. We show that GW501516 decreased phosphate and tensin homolog deleted on chromosome 10 (PTEN), a tumor suppressor known to decrease cell growth and induce apoptosis. Activation of PPARbeta/delta and phosphatidylinositol 3-kinase (PI3K)/Akt signaling was associated with inhibition of PTEN. GW501516 increased NF-kappaB DNA binding activity and p65 protein expression through activation of PPARbeta/delta and PI3K/Akt signals and enhanced the physical interactions between PPARbeta/delta and p65 protein. Conversely, inhibition of PI3K and silencing of p65 by small RNA interference (siRNA) blocked the effect of GW501516 on PTEN expression and on NSCLC cell proliferation. GW501516 also inhibited IKBalpha protein expression. Silencing of IKBalpha enhanced the effect of GW501516 on PTEN protein expression and on cell proliferation. It also augmented the GW501516-induced complex formation of PPARbeta/delta and p65 proteins. Overexpression of PTEN suppressed NSCLC cell growth and eliminated the effect of GW501516 on phosphorylation of Akt. Together, our observations suggest that GW501516 induces the proliferation of NSCLC cells by inhibiting the expression of PTEN through activation of PPARbeta/delta, which stimulates PI3K/Akt and NF-kappaB signaling. Overexpression of PTEN overcomes this effect and unveils PPARbeta/delta and PTEN as potential therapeutic targets in NSCLC. PMID- 18390838 TI - Colour and translucency of tooth-coloured orthodontic brackets. AB - The objective of this study was to determine the reflected and transmitted colours and the diffuse light transmittance of tooth-coloured brackets. Four ceramic and four plastic brands were evaluated and five brackets of each brand were tested. Reflected colour and spectral reflectance of the labial surface of the brackets were measured according to the Commission Internationale de l'Eclairage (CIE) colour scale and transmitted colour and diffuse spectral transmittance measured with a spectrophotometer. One-way analyses of variance were performed for the reflected and transmitted colour co-ordinates (CIE L*, a*, and b*) and for light transmittance according to bracket brand. The range for CIE L* (lightness) was 36.2-50.3, for a* (red-green parameter) -1.3-3.8 and for b* (yellow-blue parameter) -2.9-11.2. All these colour co-ordinates were influenced by bracket brand (P < 0.05). Diffuse light transmittance was also influenced by bracket brand and ranged from 44.9 to 75.9 per cent (P < 0.05). Colour and transmittance varied by bracket brand. Variations in optical properties influenced the aesthetic performance of the brackets and the degree of cure of the adhesive that is possible through the brackets. Further studies on the clinical implications of colour matching of tooth-coloured brackets with teeth should now be performed. PMID- 18390836 TI - Core binding factors are necessary for natural killer cell development and cooperate with Notch signaling during T-cell specification. AB - CBFbeta is the non-DNA binding subunit of the core binding factors (CBFs). Mice with reduced CBFbeta levels display profound, early defects in T-cell but not B cell development. Here we show that CBFbeta is also required at very early stages of natural killer (NK)-cell development. We also demonstrate that T-cell development aborts during specification, as the expression of Gata3 and Tcf7, which encode key regulators of T lineage specification, is substantially reduced, as are functional thymic progenitors. Constitutively active Notch or IL-7 signaling cannot restore T-cell expansion or differentiation of CBFbeta insufficient cells, nor can overexpression of Runx1 or CBFbeta overcome a lack of Notch signaling. Therefore, the ability of the prethymic cell to respond appropriately to Notch is dependent on CBFbeta, and both signals converge to activate the T-cell developmental program. PMID- 18390837 TI - First clinical use of ofatumumab, a novel fully human anti-CD20 monoclonal antibody in relapsed or refractory follicular lymphoma: results of a phase 1/2 trial. AB - Ofatumumab is a unique monoclonal antibody that targets a distinct small loop epitope on the CD20 molecule. Preclinical data show that ofatumumab is active against B-cell lymphoma/chronic lymphocytic leukemia cells with low CD20-antigen density and high expression of complement inhibitory molecules. In a phase 1/2 trial evaluating safety and efficacy of ofatumumab in relapsed or refractory follicular non-Hodgkin lymphoma (FL) grade 1 or 2, 4 dose groups of 10 patients received 4 weekly infusions of 300, 500, 700, or 1000 mg. Patients had a median of 2 prior FL therapies and 13% had elevated lactate dehydrogenase. No safety concerns or maximum tolerated dose was identified. A total of 274 adverse events were reported; 190 were judged related to ofatumumab, most occurring on the first infusion day with Common Terminology Criteria grade 1 or 2. Eight related events were grade 3. Treatment caused immediate and profound B-cell depletion, and 65% of patients reverted to negative BCL2 status. Clinical response rates ranged from 20% to 63%. Median time to progression for all patients/responders was 8.8/32.6 months, and median duration of response was 29.9 months at a median/maximum follow-up of 9.2/38.6 months. Ofatumumab is currently being evaluated in patients with rituximab-refractory FL. This trial was registered at www.clinicaltrials.gov as #NCT00092274. PMID- 18390839 TI - Satisfaction with and psychological impact of immediate and deferred breast reconstruction. AB - BACKGROUND: The present work assesses the effect of immediate breast reconstruction (IBR), deferred breast reconstruction (DBR), and no breast reconstruction on the psychological impact. PATIENTS AND METHODS: Standard questionnaires were used to determine the psychological impact suffered by patients who underwent IBR, DBR and no reconstruction, their degree of satisfaction with the results achieved, and their postprocedure opinions regarding reconstruction options. RESULTS: A total of 526 women underwent mastectomy. The response rate to the questionnaires was 71.67%. A significantly greater proportion of the women who underwent no reconstruction suffered psychological problems than those who underwent reconstruction of some type (P = 0.01). Some 94.77% of the women who underwent IBR maintained a postprocedure preference for this option; in contrast, some 87.27% of the DBR and 56.14% of the no-reconstruction patients declared a postprocedure preference for IBR. In all, 63.49% of the women who underwent reconstruction were moderately very satisfied with the aesthetic results achieved, while only 22.80% of the no-reconstruction patients declared such satisfaction (P = 0.0001). CONCLUSIONS: The women who underwent no breast reconstruction suffered more emotional problems than those who underwent a reconstruction procedure. In general, all groups reported a postprocedure preference for IBR in their questionnaire answers. The aesthetic results achieved by IBR seem to be those best accepted. PMID- 18390840 TI - Analgesic drug use and risk of epithelial ovarian cancer. AB - Analgesic use may reduce ovarian cancer risk, possibly through antiinflammatory or antigonadotropic effects. The authors conducted a population-based, case control study in Washington State that included 812 women aged 35-74 years who were diagnosed with epithelial ovarian cancer between 2002 and 2005 and 1,313 controls. Use of analgesics, excluding use within the previous year, was assessed via in-person interviews. Logistic regression was used to calculate odds ratios and 95% confidence intervals. Overall, acetaminophen and aspirin were associated with weakly increased risks of ovarian cancer. These associations were stronger after more than 10 years of use (acetaminophen: odds ratio (OR) = 1.8, 95% confidence interval (CI): 1.3, 2.6; aspirin: OR = 1.6, 95% CI: 1.1, 2.2) and were present for indications of headache, menstrual pain, and other pain/injury. Reduced risk was observed among aspirin users who began regular use within the previous 5 years (OR = 0.6, 95% CI: 0.4, 1.0) or used this drug for prevention of heart disease (OR = 0.7, 95% CI: 0.5, 1.0). These results, in the context of prior findings, do not provide compelling evidence of a true increase in risk of ovarian cancer among women who use these drugs. However, they add to the weight of evidence that, in the aggregate, provides little support for the use of analgesic drugs as chemoprevention for this disease. PMID- 18390841 TI - Association between serum trans-monounsaturated fatty acids and breast cancer risk in the E3N-EPIC Study. AB - The authors assessed the association between serum phospholipid fatty acids as biomarkers of fatty acid intake and breast cancer risk among women in the E3N Study (1989-2002), the French component of the European Prospective Investigation into Cancer and Nutrition. During an average of 7 years of follow-up, 363 cases of incident invasive breast cancer were documented among 19,934 women who, at baseline (1995-1998), had completed a diet history questionnaire and provided serum samples. Controls were randomly matched to cases by age, menopausal status at blood collection, fasting status at blood collection, date, and collection center. Serum phospholipid fatty acid composition was assessed by gas chromatography. Adjusted odds ratios for risk of breast cancer with increasing levels of fatty acids were calculated using conditional logistic regression. An increased risk of breast cancer was associated with increasing levels of the trans-monounsaturated fatty acids palmitoleic acid and elaidic acid (highest quintile vs. lowest: odds ratio = 1.75, 95% confidence interval: 1.08, 2.83; p trend = 0.018). cis-Monounsaturated fatty acids were unrelated to breast cancer risk. A high serum level of trans-monounsaturated fatty acids, presumably reflecting a high intake of industrially processed foods, is probably one factor contributing to increased risk of invasive breast cancer in women. PMID- 18390842 TI - Factor structure and correlates of the dissociative experiences scale in a large offender sample. AB - The authors examined the psychometric properties, factor structure, and construct validity of the Dissociative Experiences Scale (DES) in a large offender sample (N = 1,515). Although the DES is widely used with community and clinical samples, minimal work has examined offender samples. Participants were administered self report and interview measures, and a subsample was followed longitudinally to determine criminal and violent recidivism. The DES exhibited good psychometric properties, but an identified three-factor structure was of questionable replicability. Moreover, the DES factors displayed no evidence of differential correlates. DES total scores were correlated with trauma-related variables even after controlling for negative affectivity. Total scores were related to measures of antisocial behavior and aggression but did not predict recidivism. These findings support the reliability and construct validity of the DES in offenders but raise questions regarding the clinical utility of the DES factor scores above and beyond that of the total score. PMID- 18390843 TI - Role of FoxO1 activation in MDR1 expression in adriamycin-resistant breast cancer cells. AB - The development of multidrug resistance 1 (MDR1) can be mediated by a number of different mechanisms but elevated gene expression of MDR1 (P-glycoprotein) has often been a major cause of chemoresistance in many cancer cells. Therefore, the present study aimed to investigate the role of forkhead box-containing protein, O subfamily (FoxO), transcription factors in regulating the MDR1 gene expression. The proximal promoter region of the human MDR1 contained a putative FoxO-binding site, which partially overlapped with the enhancer/enhancer-binding protein beta binding region. Gel shift and immunoblot analysis of subcellular fractions revealed that nuclear levels of FoxO1 and its DNA-binding activity were selectively enhanced in MCF-7/ADR cells, which was reversed by a FoxO1 antibody. Reporter gene assays showed that the transcription of MDR1 gene is stimulated by FoxO1 overexpression. Moreover, both MDR1 expression and doxorubicin resistance in MCF-7/ADR cells were reversed by FoxO1 small interfering RNA (siRNA). The MDR1 expression in MCF-7/ADR cells was also inhibited by insulin, a functional FoxO1 inactivator. In conclusion, FoxO1 is a novel transcriptional activator of MDR1 and is crucial for MDR1 induction in MCF-7/ADR cells. PMID- 18390844 TI - MDM2 and p53 polymorphisms are associated with the development of hepatocellular carcinoma in patients with chronic hepatitis B virus infection. AB - A single-nucleotide polymorphism (SNP) in the promoter region of MDM2, SNP 309, is associated with hepatocellular carcinoma (HCC) in patients with chronic hepatitis C virus infection. The effect of p53 codon 72 polymorphism Arg72Pro on HCC risk remains inconsistent. This study evaluated the association of MDM2 and p53 polymorphisms with the presence and early onset of HCC in Korean patients with chronic hepatitis B virus (HBV) infection. In total, 583 consecutive patients with chronic HBV infection were classified according to the presence (n = 287) or absence (n = 296) of HCC. The MDM2 SNP 309 and p53 Arg72Pro were genotyped using restriction fragment length polymorphism method. The MDM2 G/G and p53 Pro/Pro genotypes were more frequent in HCC group than in non-HCC group (P < 0.001 and P = 0.004, respectively). Multivariate analysis for the presence of HCC revealed that the odds ratio (OR) for MDM2 G/G over T/T was 4.89 (P < 0.001) and that of p53 Pro/Pro over Arg/Arg was 3.03 (P = 0.006). Combined MDM2 G/G and p53 Pro/Pro had a synergistic effect on HCC risk, with an OR of 20.78 (P < 0.001). The mean age of tumor onset in patients with MDM2 G/G genotype was 50.9 years compared with 55.1 with T/T genotype (P = 0.018) and that with p53 Pro/Pro was 49.7 years compared with 52.9 with Arg/Arg (P = 0.040). Thus, MDM2 SNP 309 and p53 Arg72Pro are associated with the early development of HCC in Korean patients with chronic HBV infection. PMID- 18390845 TI - A review of similarities between domain-specific determinants of four health behaviors among adolescents. AB - Schools are overloaded with health promotion programs that, altogether, focus on a broad array of behavioral domains, including substance abuse, sexuality and nutrition. Although the specific content of programs varies according to the domain focus, programs usually address similar concepts: knowledge, attitudinal beliefs, social influences and skills. This apparent conceptual overlap between behaviors and programs provides opportunities for a transfer-oriented approach which will stimulate students to apply the knowledge and skills they have learned in one domain (e.g. skills for resisting tobacco use) to other domains (e.g. alcohol, sex). A requirement for such an approach is that behaviors share at least some determinants. This review addresses this issue by examining similarities between domain-specific determinants of smoking, drinking, safe sex and healthy nutrition among adolescents. Recent empirical studies and reviews were examined. The results show that the following determinants are relevant to all four behaviors: beliefs about immediate gratification and social advantages, peer norms, peer and parental modeling and refusal self-efficacy. Several other determinants have been found to relate to at least two behaviors, e.g. health risk beliefs and parental norms. These results can be used for the development of a transfer-oriented school health promotion curriculum. PMID- 18390846 TI - FLC or not FLC: the other side of vernalization. AB - Vernalization is the promotion of the competence for flowering by long periods of low temperatures such as those typically experienced during winters. In Arabidopsis, the vernalization response is, to a large extent, mediated by the repression of the floral repressor FLC, and the stable epigenetic silencing of FLC after cold treatments is essential for vernalization. In addition to FLC, other vernalization targets exist in Arabidopsis. In grasses, vernalization seems to be entirely independent of FLC. Here, the current understanding of FLC independent branches of the vernalization pathway in Arabidopsis and vernalization without FLC in grasses is discussed. This review focuses on the role of AGL19, AGL24, and the MAF genes in Arabidopsis. Interestingly, vernalization acts through related molecular machineries on distinct targets. In particular, protein complexes similar to Drosophila Polycomb Repressive Complex 2 play a prominent role in establishing an epigenetic cellular memory for cold regulated expression states of AGL19 and FLC. Finally, the similar network topology of the apparently independently evolved vernalization pathways of grasses and Arabidopsis is discussed. PMID- 18390847 TI - Expression of the nuclear gene TaF(A)d is under mitochondrial retrograde regulation in anthers of male sterile wheat plants with timopheevii cytoplasm. AB - Alterations of mitochondrial-encoded subunits of the F(o)F(1)-ATP synthase are frequently associated with cytoplasmic male sterility (CMS) in plants; however, little is known about the relationship of the nuclear encoded subunits of this enzyme with CMS. In the present study, the full cDNA of the gene TaF(A)d that encodes the putative F(A)d subunit of the F(o)F(1)-ATP synthase was isolated from the wheat (Triticum aestivum) fertility restorer '2114' for timopheevii cytoplasm based CMS. The deduced 238 amino acid polypeptide is highly similar to its counterparts in dicots and other monocots but has low homology to its mammalian equivalents. TaF(A)d is a single copy gene in wheat and maps to the short arm of the group 6 chromosomes. Transient expression of the TaF(A)d-GFP fusion in onion epidermal cells demonstrated TaF(A)d's mitochondrial location. TaF(A)d was expressed abundantly in stem, leaf, anther, and ovary tissues of 2114. Nevertheless, its expression was repressed in anthers of CMS plants with timopheevii cytoplasm. Genic male sterility did not affect its expression in anthers. The expression of the nuclear gene encoding the 20 kDa subunit of F(o) was down-regulated in a manner similar to TaF(A)d in the T-CMS anthers while that of genes encoding the 6 kDa subunit of F(o) and the gamma subunit of F(1) was unaffected. These observations implied that TaF(A)d is under mitochondrial retrograde regulation in the anthers of CMS plants with timopheevii cytoplasm. PMID- 18390848 TI - Structure-function analysis of the NB-ARC domain of plant disease resistance proteins. AB - Resistance (R) proteins in plants are involved in pathogen recognition and subsequent activation of innate immune responses. Most resistance proteins contain a central nucleotide-binding domain. This so-called NB-ARC domain consists of three subdomains: NB, ARC1, and ARC2. The NB-ARC domain is a functional ATPase domain, and its nucleotide-binding state is proposed to regulate activity of the R protein. A highly conserved methionine-histidine aspartate (MHD) motif is present at the carboxy-terminus of ARC2. An extensive mutational analysis of the MHD motif in the R proteins I-2 and Mi-1 is reported. Several novel autoactivating mutations of the MHD invariant histidine and conserved aspartate were identified. The combination of MHD mutants with autoactivating hydrolysis mutants in the NB subdomain showed that the autoactivation phenotypes are not additive. This finding indicates an important regulatory role for the MHD motif in the control of R protein activity. To explain these observations, a three-dimensional model of the NB-ARC domain of I-2 was built, based on the APAF-1 template structure. The model was used to identify residues important for I-2 function. Substitution of the selected residues resulted in the expected distinct phenotypes. Based on the model, it is proposed that the MHD motif fulfils the same function as the sensor II motif found in AAA+ proteins (ATPases associated with diverse cellular activities)-co-ordination of the nucleotide and control of subdomain interactions. The presented 3D model provides a framework for the formulation of hypotheses on how mutations in the NB ARC exert their effects. PMID- 18390849 TI - Silencing of acidic pathogenesis-related PR-1 genes increases extracellular beta (1->3)-glucanase activity at the onset of tobacco defence reactions. AB - The class 1 pathogenesis-related (PR) proteins are thought to be involved in plant defence responses, but their molecular functions are unknown. The function of PR-1 was investigated in tobacco by generating stable PR-1a-silenced lines in which other acidic PR-1 genes (PR-1b and PR-1c) were silenced. Plants lacking extracellular PR-1s were more susceptible than wild-type plants to the oomycete Phytophthora parasitica but displayed unaffected systemic acquired resistance and developmental resistance to this pathogen. Treatment with salicylic acid up regulates the PR-1g gene, encoding a basic protein of the PR-1 family, in PR-1 deficient tobacco, indicating that PR-1 expression may repress that of PR-1g. This shows that acidic PR-1s are dispensable for expression of salicylic acid dependent acquired resistances against P. parasitica and may reveal a functional overlap in tobacco defence or a functional redundancy in the PR-1 gene family. The data also show that there is a specific increase in apoplastic beta-(1-->3) glucanase activity and a decrease in beta-(1-->3)-glucan deposition in PR-1 silenced lines following activation of defence reactions. Complementation of the silencing by apoplastic treatment with a recombinant PR-1a protein largely restores the wild-type beta-(1-->3)-glucanase activity and callose phenotype. Taken together with the immunolocalization of PR-1a to sites of beta-(1-->3) glucan deposition in wild-type plants, these results are indicative of a function for PR-1a in regulation of enzymatic activity of extracellular beta-(1-->3) glucanases. PMID- 18390850 TI - Structural, biochemical, and physiological characterization of photosynthesis in two C4 subspecies of Tecticornia indica and the C3 species Tecticornia pergranulata (Chenopodiaceae). AB - Among dicotyledon families, Chenopodiaceae has the most C(4) species and the greatest diversity in structural forms of C(4). In subfamily Salicornioideae, C(4) photosynthesis has, so far, only been found in the genus Halosarcia which is now included in the broadly circumscribed Tecticornia. Comparative anatomical, cytochemical, and physiological studies on these taxa, which have near-aphyllous photosynthetic shoots, show that T. pergranulata is C(3), and that two subspecies of T. indica (bidens and indica) are C(4) (Kranz-tecticornoid type). In T. pergranulata, the stems have two layers of chlorenchyma cells surrounding the centrally located water storage tissue. The two subspecies of T. indica have Kranz anatomy in reduced leaves and in the fleshy stem cortex. They are NAD-malic enzyme-type C(4) species, with mesophyll chloroplasts having reduced grana, characteristic of this subtype. The Kranz-tecticornoid-type anatomy is unique among C(4) types in the family in having groups of chlorenchymatous cells separated by a network of large colourless cells (which may provide mechanical support or optimize the distribution of radiation in the tissue), and in having peripheral vascular bundles with the phloem side facing the bundle sheath cells. Also, the bundle sheath cells have chloroplasts in a centrifugal position, which is atypical for C(4) dicots. Fluorescence analyses in fresh sections indicate that all non-lignified cell walls have ferulic acid, a cell wall cross-linker. Structural-functional relationships of C(4) photosynthesis in T. indica are discussed. Recent molecular studies show that the C(4) taxa in Tecticornia form a monophyletic group, with incorporation of the Australian endemic genera of Salicornioideae, including Halosarcia, Pachycornia, Sclerostegia, and Tegicornia, into Tecticornia. PMID- 18390851 TI - Selection on amino acid substitutions in Arabidopsis. AB - Studies of nucleotide diversity have found an excess of low-frequency amino acid polymorphisms segregating in Arabidopsis thaliana, suggesting a predominance of weak purifying selection acting on amino acid polymorphism in this inbreeding species. Here, we investigate levels of diversity and divergence at synonymous and nonsynonymous sites in 6 circumpolar populations of the outbreeding Arabidopsis lyrata and compare these results with A. thaliana, to test for differences in mutation and selection parameters across genes, populations, and species. We find that A. lyrata shows an excess of low-frequency nonsynonymous polymorphisms both within populations and species wide, consistent with weak purifying selection similar to the patterns observed in A. thaliana. Furthermore, nonsynonymous polymorphisms tend to be more restricted in their population distribution in A. lyrata, consistent with purifying selection preventing their geographic spread. Highly expressed genes show a reduced ratio of amino acid to synonymous change for both polymorphism and fixed differences, suggesting a general pattern of stronger purifying selection on high-expression proteins. PMID- 18390852 TI - Low ratios of sodium to potassium in the serum of 238 dogs. AB - Serum sodium:potassium (Na:K) ratios are often reported in biochemical studies of dogs, although their value has not been assessed. The aims of this study were to identify diseases associated with a low Na:K ratio in dogs and to compare their prevalence with the prevalence in dogs from the same referral hospital with normal Na:K ratios. A total of 238 dogs with a Na:K ratio less than 27 were identified from medical records. Sample contamination with edta was suspected in 74 cases (31 per cent) and these and two cases that had been supplemented with potassium were removed from the analysis. The remaining 162 cases and 147 control dogs were divided into five categories depending on the organ system affected. Among the cases there were significantly more in the endocrine category than among the control dogs. Hypoadrenocorticism was the most single common cause of a low Na:K ratio and affected 27 (16.7 per cent) of the cases. Other clinical problems associated with low Na:K ratios included different urogenital, cardiorespiratory and gastrointestinal diseases. PMID- 18390853 TI - Dog-human and dog-dog interactions of 260 dog-owning households in a community in Cheshire. AB - This study investigated the nature and frequency of the contacts that occur between dogs, and between dogs and people, by means of a questionnaire survey of 260 dog-owning households in a community in Cheshire, uk. The contacts were highly variable and were affected by the size, sex and age of the dog, individual dog behaviours, human behaviours and human preferences in the management of the dog. A number of situations were identified that may be important in relation to zoonoses, including sleeping areas, playing behaviours, greeting behaviours, food sources, walking, disposal of faeces, veterinary preventive treatment and general hygiene. PMID- 18390854 TI - Quantitative and qualitative assessments of intraoral lesions in 180 small herbivorous mammals. AB - Between 2002 and 2005 210 rabbits, 257 guinea pigs and 123 chinchillas were examined; oral disease was diagnosed in 38.1 per cent of the rabbits, 23.4 per cent of the guinea pigs and 32.5 per cent of the chinchillas. In the rabbits, the maxillary right P3, mandibular right P3, P4 and M1 and mandibular left P3, P4, and M1 were the teeth most frequently affected; in the guinea pigs, the mandibular right P4 and both the mandibular left incisor and P4 were most often affected; and in the chinchillas the maxillary right P4 and M1 and maxillary left P4 were most often affected. The incisors and first mandibular cheek teeth of the guinea pigs were significantly more often affected than those in the chinchillas or rabbits. Bilateral lesions of the maxillary cheek teeth were significantly more common in the chinchillas. The highest incidence of gingival hyperplasia was recorded in the rabbits. The incidence of gingival erosions and gingivitis were significantly higher in the chinchillas. PMID- 18390855 TI - Fatal necrotising fasciitis and myositis in a cat associated with Streptococcus canis. AB - Necrotising fasciitis and necrotising myositis are rare but serious life threatening conditions reported mainly in human beings and dogs. Most cases to date have been caused by beta-haemolytic streptococci of Lancefield groups A, B, C or G. Necrotising fasciitis has been reported only twice in cats and necrotising myositis has never been described. This paper describes a fatal case of necrotising fasciitis and myositis with pneumonia and septicaemia in a nine year-old cat. The cat had been undergoing treatment for a suspected tear of the cranial cruciate ligament, but on the seventh day of treatment it suddenly deteriorated and died. On postmortem examination, there was an area of hair loss from its left hindlimb and discoloration of the underlying fascia and biceps femoris muscle. Severe necrotising fasciitis and myositis, with numerous intralesional Gram-positive coccoid bacteria, was diagnosed histologically. Other findings included necrotising pneumonia, pleuritis, focal encephalitis, myocarditis and nephritis. Culture of the affected tissues yielded a pure, heavy growth of Streptococcus canis. PMID- 18390856 TI - Interferon-gamma test for feline tuberculosis. PMID- 18390857 TI - B cell lymphoma causing splenomegaly in a Sundevall's jird (Meriones crassus subspecies perpallidus). PMID- 18390859 TI - RCVS disciplinary procedures. PMID- 18390860 TI - Bluetongue virus antibodies in wild red deer in southern Belgium. PMID- 18390861 TI - Bluetongue vaccination roadshows. PMID- 18390862 TI - Psoroptic mange in cattle in south Wales. PMID- 18390864 TI - Use of human spironolactone in dogs with heart failure. PMID- 18390865 TI - Impact of functional limitations and medical comorbidity on subsequent weight changes and increased depressive symptoms in older adults. AB - OBJECTIVE: The primary goal of this study was to determine the effect of the onset of major medical comorbidity and functional decline on subsequent weight change and increased depressive symptoms. METHODS: The sample included a prospective cohort of 53 to 63 year olds (n = 10,150) enrolled in the Health and Retirement Study. Separate lagged covariate models for men and women were used to study the impact of functional decline and medical comorbidity on subsequent increases in depressive symptoms and weight change 2 years later. RESULTS: Functional decline and medical comorbidity were individual predictors of subsequent weight changes but not increased depressive symptoms. Most specific incident medical comorbidities or subtypes of functional decline predicted weight changes in both directions. DISCUSSION: The elevated risk of weight gain subsequent to functional decline or onset of medical comorbidities may require the receipt of preventive measures to reduce further weight-related complications. PMID- 18390867 TI - Sub-aortic mitral impact lesion depicted by delayed enhancement cardiac magnetic resonance imaging. PMID- 18390866 TI - Promoting the effective translation of the framework convention on tobacco control: a case study of challenges and opportunities for strategic communications in Mexico. AB - The World Health Organization Framework Convention on Tobacco Control (WHO-FCTC) promotes the implementation of best-practices tobacco control policies at a global scale. This article describes features of the sociocultural and political economic context of Mexico that pose challenges and opportunities to the effective translation of WHO-FCTC policies there. It also considers how strategic communication efforts may advance these policies by framing their arguments in ways that resonate with prevalent values, understandings, and concerns. A focus on a smoke-free policy illustrates barriers to policy compliance, including how similar issues have been overcome among Latino populations in California. Overall, this article aims to lay the foundation for comparative research from policy uptake to impact so that the scientific evidence base on tobacco control policies includes examination of how context moderates this process. PMID- 18390868 TI - Atherogenic dyslipidaemia but not total- and high-molecular weight adiponectin are associated with the prognostic outcome in patients with coronary heart disease. AB - AIMS: Adiponectin is closely related to atherogenic dyslipidaemia and may be a clinical important mediator of recurrent coronary heart disease (CHD). However, studies with emphasis on secondary disease prevention are rare. We report data from a prospective study investigating the prognostic value of adiponectin, its high-molecular weight (HMW) form, and of markers of lipid metabolism in patients after their first acute CHD event. METHODS AND RESULTS: We measured baseline total- and HMW-adiponectin in 1051 patients aged 30-70 years with incident CHD and a prospective follow-up was conducted [median: 56.6 months (interquartile range: 53.2; 57.5)]. During this period, 95 patients (incidence: 22.3/1000 patient years) experienced a secondary cardiovascular disease (CVD) event. After adjustment by Cox proportional hazard models, neither total- nor HMW-adiponectin was associated with secondary CVD events. In contrast, LDL-cholesterol and markers of atherogenic dyslipidaemia were independently associated with secondary CVD events (relative risk per unit increase: LDL-cholesterol: 1.54; 95%CI 1.18 2.01; P = 0.001, triglycerides: 1.58; 95%CI 1.31-1.90; P < 0.0001 and HDL cholesterol: 0.34; 95%CI 0.14-0.79; P = 0.01). CONCLUSION: Measurement of total- and HMW-adiponectin may add no significant value to risk stratifications in patients with incident CHD. In contrast, approaching atherogenic dyslipidaemia may represent a promising target in secondary prevention programs for high-risk patients. PMID- 18390870 TI - Osteosarcoma in left atrium treated with autotransplant. PMID- 18390869 TI - Long-term survival in patients undergoing cardiac resynchronization therapy: the importance of performing atrio-ventricular junction ablation in patients with permanent atrial fibrillation. AB - AIMS: To investigate the effects of cardiac resynchronization therapy (CRT) on survival in heart failure (HF) patients with permanent atrial fibrillation (AF) and the role of atrio-ventricular junction (AVJ) ablation in these patients. METHODS AND RESULTS: Data from 1285 consecutive patients implanted with CRT devices are presented: 1042 patients were in sinus rhythm (SR) and 243 (19%) in AF. Rate control in AF was achieved by either ablating the AVJ in 118 patients (AVJ-abl) or prescribing negative chronotropic drugs (AF-Drugs). Compared with SR, patients with AF were significantly older, more likely to be non-ischaemic, with higher ejection fraction, shorter QRS duration, and less often received ICD back-up. During a median follow-up of 34 months, 170/1042 patients in SR and 39/243 in AF died (mortality: 8.4 and 8.9 per 100 person-year, respectively). Adjusted hazard ratios were similar for all-cause and cardiac mortality [0.9 (0.57-1.42), P = 0.64 and 1.00 (0.60-1.66) P = 0.99, respectively]. Among AF patients, only 11/118 AVJ-abl patients died vs. 28/125 AF-Drugs patients (mortality: 4.3 and 15.2 per 100 person-year, respectively, P < 0.001). Adjusted hazard ratios of AVJ-abl vs. AF-Drugs was 0.26 [95% confidence interval (CI) 0.09 0.73, P = 0.010] for all-cause mortality, 0.31 (95% CI 0.10-0.99, P = 0.048) for cardiac mortality, and 0.15 (95% CI 0.03-0.70, P = 0.016) for HF mortality. CONCLUSION: Patients with HF and AF treated with CRT have similar mortality compared with patients in SR. In AF, AVJ ablation in addition to CRT significantly improves overall survival compared with CRT alone, primarily by reducing HF death. PMID- 18390871 TI - Sudden cardiac death due to acute right heart failure in a patient with arrhythmogenic right ventricular dysplasia and acute myocardial infarction. PMID- 18390872 TI - A rare cause of cardiac tumour: an Erdheim-Chester disease with cardiac involvement co-existing with an intracerebral Langerhans cell histiocytosis. PMID- 18390873 TI - Novel biomarkers--the long march from bench to bedside. PMID- 18390874 TI - Automatic R-wave and impedance testing with the modern patient alert system to reduce inappropriate implantable cardioverter defibrillator shocks due to lead fracture. AB - A 62-year-old man was afflicted with implantable cardioverter defibrillator (ICD) shocks during sinus rhythm. Stored ICD data revealed that sensing of noise due to fracture of the ventricular lead triggered the delivery of shocks. Since the lead fracture developed suddenly, it is suggested that close, early attention should be paid to the potential of such events during follow-up of ICD leads. PMID- 18390875 TI - Long-term endurance sport practice increases the incidence of lone atrial fibrillation in men: a follow-up study. AB - AIMS: The aim of this study is to determine the incidence of lone atrial fibrillation (LAF) in males according to sport practice and to identify possible clinical markers related to LAF among marathon runners. METHODS AND RESULTS: A retrospective cohort study was designed. A group of marathon runners (n = 252) and a population-based sample of sedentary men (n = 305) recruited in 1990-92 and 1994-96, respectively, were contacted in 2002-03 and invited to attend an outpatient clinic to identify suggestive symptoms of having experienced an arrhythmia requiring medical attention. In those with suggestive symptoms of atrial fibrillation, medical records were reviewed. Finally, LAF was diagnosed on the basis of the presence of atrial fibrillation in an electrocardiographic recording. In the group of marathon runners, an echocardiogram was performed at inclusion and at the end of the study. The annual incidence rate of LAF among marathon runners and sedentary men was 0.43/100 and 0.11/100, respectively. Endurance sport practice was associated with a higher risk of incident LAF in the multivariate age- and blood pressure-adjusted Cox regression models (hazard ratio = 8.80; 95% confidence interval: 1.26-61.29). In the group of marathon runners, left atrial inferosuperior diameter and left atrial volume were both associated with a higher risk of incident LAF. CONCLUSION: Long-term endurance sport practice is associated with a higher risk of symptomatic LAF in men. This risk is associated with a larger left atrial inferosuperior diameter and volume in physically active subjects. PMID- 18390876 TI - Simple primary structure, complex turnover regulation and multiple roles of hyaluronan. AB - Hyaluronan is a major macromolecular polysaccharide component of the extra cellular matrix that confers structural frameworks for cells. Despite its relatively simple chemical composition, hyaluronan mediates many other important functional aspects including signalling activity during embryonic morphogenesis, cellular regeneration and wound healing. Abnormalities in hyaluronan metabolism have been implicated in many diseases, such as inflammatory disorders, cardiovascular diseases and cancer. To date, it has become increasingly clear that hyaluronan production in vertebrates is tightly regulated by three hyaluronan synthases and that hyaluronan catabolism is regulated by an enzymatic degradation reaction involving several hyaluronidases. Together, these discoveries have provided key insights into the physiological roles of hyaluronan and a deeper understanding of the mechanisms underlying altered hyaluronan turnover in diseases. The central aim of this review article is therefore to highlight the multiple roles of hyaluronan in physiological and pathological states via its complex turnover regulation. PMID- 18390877 TI - The nucleocapsid protein of SARS-CoV induces transcription of hfgl2 prothrombinase gene dependent on C/EBP alpha. AB - Fibrin deposition was universal in the lungs of SARS patients and fgl2 prothrombinase gene, a novel procoagulant, was demonstrated to express highly in a clinically relevant SARS model. To investigate whether and which structural protein of SARS-CoV induced transcription of hfgl2 prothrombinase gene, three eukaryotic expression plasmids expressing nucleocapsid protein (N), membrane protein (M) and spike protein 2 (S2) of SARS-CoV were co-transfected with hfgl2 promoter luciferase-reporter plasmids and beta-galactosidase plasmid in CHO cells, respectively. M, N and S2 protein of SARS-CoV were detected by western blotting and immunohistochemistry analysis. Further assays demonstrated that expression of hfgl2 gene was related with N protein, but not with M or S2 protein in THP-1 cells and Vero cells. N protein significantly induced functional procoagulant activity in comparison with control group. Luciferase assay showed that N protein of SARS-CoV could activate the transcription of hfgl2 promoter compared with the pcDNA3.1 empty vector. Site-directed mutagenesis and EMSA assay further demonstrated that transcription factor C/EBP alpha band with its cognate cis-element in hfgl2 promoter. The results showed that N protein of SARS-CoV induced hfgl2 gene transcription dependent on the transcription factor C/EBP alpha, which maybe contribute to the development of thrombosis in SARS. PMID- 18390878 TI - Use of baseline and updated information on alcohol intake on risk for breast cancer: importance of latency. AB - BACKGROUND: Alcohol intake has been shown to be associated with an increased risk for breast cancer. In the analysis of longitudinal prospective cohort studies, however, the analysis of repeated measurements of alcohol intake might not be straightforward. METHODS: In this analysis of the Copenhagen City Heart Study, in which alcohol intake was measured four times, 9318 Danish women with no previous diagnosis of cancer were followed for breast cancer for 27 years, from 1976 to 2002. During follow-up, breast cancer was diagnosed in 476 women. RESULTS: The association between alcohol intake at first measurement (baseline alcohol intake) and breast cancer was positive and approximately linear. When alcohol intake was updated during follow-up, no association was observed between breast cancer and alcohol intake. It is suggested that this difference in results may be attributable to long latency time between alcohol intake and breast cancer occurrence, because a markedly increased risk was estimated on the basis of direct lagging of risk time. CONCLUSIONS: Our results support the hypothesis that baseline alcohol intake is more strongly associated with breast cancer risk than updated intake, and we suggest that this is due to the long latency between alcohol intake and breast cancer. PMID- 18390879 TI - UniProtJAPI: a remote API for accessing UniProt data. AB - Programmatic access to the UniProt Knowledgebase (UniProtKB) is essential for many bioinformatics applications dealing with protein data. We have created a Java library named UniProtJAPI, which facilitates the integration of UniProt data into Java-based software applications. The library supports queries and similarity searches that return UniProtKB entries in the form of Java objects. These objects contain functional annotations or sequence information associated with a UniProt entry. Here, we briefly describe the UniProtJAPI and demonstrate its usage. PMID- 18390880 TI - Risk of future sickness absence in frequent and long-term absentees. AB - BACKGROUND: Prior absence is an important predictor for sickness absence, but little is known about the recurrence among frequent and/or long-term absentees, over a longer period of time. AIM: To monitor sickness absence among frequent and long-term absentees in order to investigate their risk of recurrent absence. METHODS: Longitudinal cohort study in employees working in three large Dutch postal and telecommunications companies. In the first year of study, we distinguished employees who were absent four times or more (frequent absence), employees who were absent for >or=6 weeks (long-term absence), combined frequent and long-term absence and a reference population. The absence rates in these groups were followed-up for 4 years. RESULTS: The study population (n = 53,990) comprised 4126 frequent absentees, 3585 long-term absentees, 979 combined frequent and long-term absentees and a reference population (n = 45,300). Frequent absentees had a higher risk of recurrent frequent absence when compared to the reference population, with rate ratios (RR) amounting to 4.9 [95% confidence interval (CI) 4.7-5.1] in men and 3.2 (95% CI 3.0-3.4) in women. They also had a higher risk of developing long-term absence: RR = 1.9 (95% CI 1.8-2.0) in men and 1.5 (95% CI 1.4-1.6) in women. Long-term absentees had high risk of recurrence: RR = 1.9 (95% CI 1.8-2.0) in men and RR = 1.4 (95% CI 1.3-1.5) in women. CONCLUSIONS: Employees with prior frequent and/or long-term absence were at risk of recurrent absence. Frequent absence was a prognostic factor predicting future long-term absence. PMID- 18390881 TI - Interactions of B-class complex proteins involved in tepal development in Phalaenopsis orchid. AB - In our previous studies, we identified four DEFICIENS (DEF)-like genes and one GLOBOSA (GLO)-like gene involved in floral organ development in Phalaenopsis equestris. Revealing the DNA binding properties and protein-protein interactions of these floral homeotic MADS-box protein complexes (PeMADS) in orchids is crucial for the elucidation of the unique orchid floral morphogenesis. In this study, the interactome of B-class PeMADS proteins was assayed by the yeast two hybrid system (Y2H) and glutathione S-transferase (GST) pull-down assays. Furthermore, the DNA binding activities of these proteins were assessed by using electrophoretic mobility shift assay (EMSA). All four DEF-like PeMADS proteins interacted individually with the GLO-like PeMADS6 in Y2H assay, yet with different strengths of interaction. Generally, the PeMADS3/PeMADS4 lineage interacted more strongly with PeMADS6 than the PeMADS2/PeMADS5 lineage did. In addition, independent homodimer formation for both PeMADS4 (DEF-like) and PeMADS6 (GLO-like) was detected. The protein-protein interactions between pairs of PeMADS proteins were further confirmed by using a GST pull-down assay. Furthermore, both the PeMADS4 homodimer and the PeMADS6 homodimer/homomultimer per se were able to bind to the MADS-box protein-binding motif CArG. The heterodimeric complexes PeMADS2-PeMADS6, PeMADS4-PeMADS6 and PeMADS5-PeMADS6 showed CArG binding activity. Taken together, these results suggest that various complexes formed among different combinations of the five B-class PeMADS proteins may increase the complexity of their regulatory functions and thus specify the molecular basis of whorl morphogenesis and combinatorial interactions of floral organ identity genes in orchids. PMID- 18390882 TI - Time-kill studies investigating the killing activity of caspofungin against Candida dubliniensis: comparing RPMI-1640 and antibiotic medium 3. AB - OBJECTIVES: We evaluated the in vitro activity of caspofungin against Candida dubliniensis strains using MIC and minimum fungicidal concentration (MFC) measurements and time-kill methodology. METHODS: We used six C. dubliniensis clinical isolates and the CD 36 type strain. MICs and MFCs of caspofungin were determined using the standard broth microdilution method with normal (10(3) cells/mL) and elevated (10(5) cells/mL) starting inocula in RPMI-1640 and antibiotic medium 3 (AM3). MIC was determined after 24 h, and plating for MFC determination was performed after 48 h. In time-kill tests, all strains were tested at 0.06-16 mg/L caspofungin concentrations in RPMI-1640 and AM3. RESULTS: In RPMI-1640, the MIC range was 0.06-8 mg/L. Trailing growth was observed regardless of the starting inoculum after 48 h, but not after 24 h. In AM3 regardless of starting inoculum, MICs were 0.03 mg/L. After 48 h, trailing was not detected; two isolates grew at a concentration of 8 mg/L using 10(5) cells/mL as the starting inoculum [paradoxical growth (PG)]. All MFCs in RPMI-1640 and AM3 were >8 and < or =0.12 mg/L, respectively. In AM3, all but a single isolate showed PG in the MFC tests. Time-kill tests confirmed the results obtained by MFC tests both in RPMI-1640 and AM3. CONCLUSIONS: In vitro activity of caspofungin against C. dubliniensis depended on the starting inoculum and medium used. Using AM3 eliminated trailing from MIC determinations but not PG in MIC, MFC and time kill tests. PMID- 18390883 TI - Predicting pathogens causing ventilator-associated pneumonia using a Bayesian network model. AB - BACKGROUND: We previously validated a Bayesian network (BN) model for diagnosing ventilator-associated pneumonia (VAP). Here, we report on the performance of the model to predict microbial causes of VAP and to select antibiotics. METHODS: Pathogens were grouped into seven categories based upon the antibiotic susceptibility and epidemiological characteristics. Colonization of the upper respiratory tract was modelled in the BN and depended--in additional steps--on (i) duration of admission and ventilation, (ii) previous culture results and (iii) previous antibiotic use. A database with 153 VAP episodes and their microbial causes was used as reference standard. Appropriateness of antibiotic prescription, with fixed choices for pathogens predicted, was determined. RESULTS: One hundred and seven VAP episodes were monobacterial and 46 were caused by two pathogens. Using duration of admission and ventilation only, areas under the receiver operating curve (AUC) ranged from 0.511 to 0.772 for different pathogen groups, and model predictions significantly improved when adding information on culture results, but not when adding information on antibiotic use. The best performing model (with all information) had AUC values ranging from 0.859 for Acinetobacter spp. to 0.929 for Streptococcus pneumoniae. With this model, 91 (85%) and 29 (63%) of all pathogen groups were correctly predicted for monobacterial and polymicrobial VAP, respectively. With fixed antibiotic choices linked to pathogen groups, 92% of all episodes would have been treated appropriately. CONCLUSIONS: The BN models' performance to predict pathogens causing VAP improved markedly with information on colonization, resulting in excellent pathogen prediction and antibiotic selection. Prospective external validation is needed. PMID- 18390884 TI - Dual-targeting properties of the 3-aminopyrrolidyl quinolones, DC-159a and sitafloxacin, against DNA gyrase and topoisomerase IV: contribution to reducing in vitro emergence of quinolone-resistant Streptococcus pneumoniae. AB - OBJECTIVES: DC-159a (a novel quinolone) and sitafloxacin (DU-6859a) are structurally related quinolones, bearing a 3-aminopyrrolidyl substitution. We investigated the relationship between the target preferences of these 3 aminopyrrolidyl quinolones, in vitro potencies and emergence of quinolone resistant mutants in Streptococcus pneumoniae, compared with other quinolones. METHODS: MICs, resistance frequencies and mutant prevention concentrations (MPCs) were determined using quinolone-susceptible strains and first-step parC mutant strains of S. pneumoniae. Target preferences were tested by the following two methods: antibacterial activities against gyrA or parC mutants and in vitro enzyme assays for the determination of 50% inhibition (IC(50)) values. RESULTS: DC-159a and sitafloxacin exhibited potent antibacterial activities, low frequencies of mutant selection, low MPCs and narrow mutant selection windows against both quinolone-susceptible strains and first-step parC mutants of S. pneumoniae, compared with gatifloxacin, moxifloxacin and other quinolones tested. DC-159a and sitafloxacin showed relatively low MIC ratios against single gyrA or parC mutants relative to the wild-type strain and low IC(50) ratios against DNA gyrase and topoisomerase IV. CONCLUSIONS: DC-159a and sitafloxacin demonstrated a more balanced dual-targeting activity than gatifloxacin, moxifloxacin and other quinolones tested. In addition, DC-159a and sitafloxacin have a lower propensity for selecting first- and second-step resistant mutants. PMID- 18390886 TI - Shaping the shoot: the relative contribution of cell number and cell shape to variations in internode length between parent and hybrid apple trees. AB - Genetic control of plant size and shape is a promising perspective, particularly in fruit trees, in order to select desirable genotypes. A recent study on architectural traits in an apple progeny showed that internode length was a highly heritable character. However, few studies have been devoted to internode cellular patterning in dicotyledonous stems, and the interplay between the two elementary cell processes that contribute to their length, i.e. cell division and elongation, is not fully understood. The present study aimed at unravelling their contributions in the genetic variation of internode length in a selection of F(1) and parent genotypes of apple tree, by exploring the number of cells and cell shape within mature internodes belonging to the main axes. The results highlighted that both the variables were homogeneous in samples collected either along a sagital line or along the pith width, and suggest that cell lengthening was homogeneous during internode development. They allowed the total number of cells to be estimated on the internode scale and opened up new perspectives for simplifying tissue sampling procedures for further investigations. Differences in internode length were observed between the genotypes, in particular between the parents, and partly resulted from a compensation between cell number and cell length. However, genetic variations in internode length primarily involved the number of cells, while cell length was more secondary. These results argue for an interplay between cellular and organismal control of internode shape that may involve the rib meristem. PMID- 18390885 TI - Clinically validated mutation scores for HIV-1 resistance to fosamprenavir/ritonavir. AB - BACKGROUND: We developed clinically relevant genotypic scores for resistance to fosamprenavir/ritonavir in HIV-1 protease inhibitor (PI)-experienced patients. METHODS: PI-experienced patients with virological failure receiving fosamprenavir/ritonavir as the sole PI for at least 3 months and with detectable fosamprenavir plasma levels were included. The impact of baseline protease mutations on virological response (VR, i.e. decrease in plasma HIV-1 RNA between baseline and month 3) was analysed using the Mann-Whitney test. Mutations with prevalence >10% and P value <0.10 were retained. The Jonckheere-Terpstra test was used to select the combination of mutations most strongly associated with VR. The association between score and VR was assessed by multivariate backward regression. RESULTS: In the 73 patients included, the median baseline HIV-1 RNA was 4.6 log(10) copies/mL (range: 2.7-6.9) and the mean decrease at month 3 was 1.07 +/- 1.40 log(10) copies/mL. Ninety per cent of the patients were infected by HIV-1 subtype B variants. Two fosamprenavir/ritonavir mutation scores were constructed: score A (L10F/I/V + L33F + M36I + I54L/M/V/A/T/S + I62V + V82A/F/C/G + I84V + L90M) was based only on mutations associated with a worse VR, whereas score B (L10FIV + L33F + M36I + I54L/M/V/A/T/S + A71V - V77I - N88S + L90M) also took into account favourable mutations. Both scores were independent predictors of VR, however, co-administration of tenofovir was associated with a worse VR and the presence of the N88S protease mutation and co-administration of enfuvirtide with a better VR. CONCLUSIONS: These clinically validated mutation scores should be of interest for the clinical management of PI-experienced patients. The fosamprenavir/ritonavir score A was introduced in the 2006 ANRS algorithm along with isolated mutations I50V and V32I + I47V. PMID- 18390887 TI - Dissection of the regulatory mechanism of a heat-shock responsive promoter in Haloarchaea: a new paradigm for general transcription factor directed archaeal gene regulation. AB - Multiple general transcription factors (GTFs), TBP and TFB, are present in many haloarchaea, and are deemed to accomplish global gene regulation. However, details and the role of GTF-directed transcriptional regulation in stress response are still not clear. Here, we report a comprehensive investigation of the regulatory mechanism of a heat-induced gene (hsp5) from Halobacterium salinarum. We demonstrated by mutation analysis that the sequences 5' and 3' to the core elements (TATA box and BRE) of the hsp5 promoter (P(hsp5)) did not significantly affect the basal and heat-induced gene expression, as long as the transcription initiation site was not altered. Moreover, the BRE and TATA box of P(hsp5) were sufficient to render a nonheat-responsive promoter heat-inducible, in both Haloferax volcanii and Halobacterium sp. NRC-1. DNA-protein interactions revealed that two heat-inducible GTFs, TFB2 from H. volcanii and TFBb from Halobacterium sp. NRC-1, could specifically bind to P(hsp5) likely in a temperature-dependent manner. Taken together, the heat-responsiveness of P(hsp5) was mainly ascribed to the core promoter elements that were efficiently recognized by specific heat-induced GTFs at elevated temperature, thus providing a new paradigm for GTF-directed gene regulation in the domain of Archaea. PMID- 18390888 TI - Accuracy of soluble human leukocyte antigen-G for predicting pregnancy among women undergoing infertility treatment: meta-analysis. AB - BACKGROUND: There have been concerns about validity and accuracy of the measurement of sHLA-G in embryo culture supernatants. In this systematic review, we quantified the diagnostic accuracy of sHLA-G for predicting the ability to achieve clinical pregnancy in women who are undergoing infertility treatment. METHODS: Medline and Embase were searched up to 7 September 2007, for full English and non-English articles concerning cohort studies evaluating sHLA-G in embryo culture for predicting clinical pregnancy in women undergoing IVF and ICSI. RESULTS: Eleven studies including 1813 patients met our inclusion criteria. In the individual studies, sensitivity ranged from 0.01 to 0.97, specificity from 0.18 to 0.98, the positive likelihood ratio from 0.34 to 3.21 and the negative likelihood ratio from 0.08 to 1.01. These values were highly heterogeneous with, in each case, I(2) values of >75%, and P-values for the Q statistic of <0.001, arguing against generating a pooled estimate for these diagnostic test properties. The diagnostic odds ratios (DORs) ranged from 0.92 to 24.82 in the individual studies with an I(2) value of 49% indicating moderate heterogeneity. Therefore, the meta-analysis combined the logs of the DORs, which are derived from sensitivity and specificity. A random-effects model yielded a summary DOR of 4.38 (95% CI, 2.93-6.55), consistent with modest diagnostic accuracy. Interestingly, an a priori defined subgroup analysis restricted to six studies with good quality embryos showed a better diagnostic performance with a DOR of 12.67 (95% CI, 3.66-43.80) to predict the ability to achieve clinical pregnancy in women undergoing infertility treatment. CONCLUSIONS: Further research is needed with single-embryo culture, single-embryo transfer and highly sensitive detection techniques to determine the potential application of measuring sHLA-G in culture supernatant. PMID- 18390889 TI - The great escape--myofibroblasts in fibrosis and the immune system. PMID- 18390891 TI - Transforming growth factor-beta1 is associated with kidney damage in patients with essential hypertension: renoprotective effect of ACE inhibitor and/or angiotensin II receptor blocker. AB - BACKGROUND: Evidence suggests that transforming growth factor-beta1 (TGF-beta(1)) is associated with target organ damage in hypertension. This study aimed to investigate the relationship between TGF-beta(1) levels and kidney damage and renoprotective effects of angiotensin-converting enzyme inhibitor and/or angiotensin II type 1 receptor blocker in patients with essential hypertension (EH). METHODS: A total of 156 patients with EH were enrolled and grouped according to albumin-to-creatinine ratio (ACR). Of these, 90 patients with EH underwent a 12-week antihypertensive trial with administration of benazepril, valsartan or both. Serum TGF-beta(1), plasma angiotensin (Ang) II and urinary albumin were quantified by immunoassays. RESULTS: Serum TGF-beta1, plasma Ang II and ACR were highly elevated in patients with EH (P < 0.01). There was a positive correlation between serum TGF-beta1 levels and ACR (r = 0.53, P < 0.01). Significant decreases in TGF beta1 and ACR were obtained in all groups at the end of 12-week antihypertensive therapy compared to the baseline values, with the combined group to a greater extent (P < 0.01). Plasma Ang II levels were significantly decreased in the benazepril group but increased in the valsartan group (P < 0.05) while no significant change was observed in the combined group. CONCLUSIONS: TGF-beta(1) is highly elevated and strongly associated with urinary albumin excretion in patients with EH. Treatment with benazepril or valsartan attenuates serum TGF-beta(1) levels and microalbuminuria with the combined therapy receiving the greater effect. TGF-beta(1) could be a potential surrogate marker in monitoring the development and progression of kidney damage in EH. PMID- 18390890 TI - Influences of haemodialysis on the binding sites of human serum albumin: possibility of an efficacious administration plan using binding inhibition. AB - BACKGROUND: We have studied the possibility that low-dose treatment utilizing the inhibition that may occur between two drugs at the same site of human serum albumin (HSA) improves the pharmacological effects. The purpose is to elucidate the differences in the binding capacities of sites I and II of HSA between pre haemodialysis (HD) and post-HD in patients with end-stage renal disease. METHODS: We evaluated free fractions of site probes, (14)C-warfarin (site I) and (14)C diazepam (site II), by ultrafiltration in serum between pre-HD and post-HD. To investigate effects on the binding capacities of HSA sites, free fractions of site probes were calculated from the radioactivities measured with a liquid scintillation counter. Endogenous uraemic toxins, 3-carboxy-4-methyl-5-propyl-2 furanpropionate (CMPF), indoxyl sulphate (IS) and hippurate (HA), were determined by HPLC. Free fatty acid (FFA) as an endogenous substance was determined with an automatic multi-item simultaneous analyser. RESULTS: The concentrations of HSA and FFA increased significantly (post-HD/pre-HD ratio: 1.18 +/- 0.10, 5.46 +/- 4.91), the concentrations of IS and HA decreased significantly (post-HD/pre-HD ratio: 0.69 +/- 0.10, 0.33 +/- 0.15) and CMPF concentrations did not alter significantly (post-HD/pre-HD ratio: 0.97 +/- 0.12, P = 0.471). The free fractions of (14)C-warfarin decreased in all 14 patients at site I at post-HD compared to pre-HD (post-HD/pre-HD ratio: 0.59 +/- 0.13). The free fractions of (14)C-diazepam at site II remarkably decreased in 10 of 14 patients (post-HD/pre HD ratio: 0.61 +/- 0.17) and unexpectedly increased in 4 (post-HD/pre-HD ratio: 1.08 +/- 0.06) post-HD compared to pre-HD. In these four patients, when we investigated the influences of these variation factors on the reduction of the binding capacities of site II, [FFA]/[HSA] increased significantly post-HD, compared to pre-HD (post-HD/pre-HD ratio: 6.91 +/- 6.58). ([FFA]/[HSA] ratios of the 4 patients were from 1.22 to 3.55, the highest for the 14 patients post-HD, but the ratios of the other 10 were below 1.2 post-HD.) CONCLUSION: The binding capacity of site II was unexpectedly decremented by the effects of the remarkable elevation of FFA. Therefore, monitoring the binding capacity of site II in HD is important for patients with end-stage renal disease in the efficacious administration plan using the binding inhibition of HSA. PMID- 18390892 TI - The detection of advanced chronic kidney disease by surveillance of elevated plasma creatinines--a five-year experience. AB - BACKGROUND: The late referral of patients with advanced chronic renal failure to a nephrologist is multifactorial but also compromises the preparations for dialysis and is prejudicial to their survival on dialysis. Measures that prompt or hasten referral, will allow preparation for dialysis, control of complications and treatment of comorbid conditions. METHODS: In June 2000, a programme was initiated to provide surveillance of plasma creatinines >300 micromol/L on all laboratory requests from general practitioners (GPs) and hospital clinicians in Ayrshire and Arran Health Board in southwest Scotland. Patients already known to the nephrologists were excluded. Results were regularly reviewed and further excluded if the creatinine fell or the patient died. For the remainder, a standard letter was sent to the requesting clinician suggesting renal referral if appropriate. This was to act as a prompt to the general practitioner or hospital clinician. For those referred over the 5-year period, the outcome was analysed in January 2007. RESULTS: In the first 5 years (June 2000-June 2005) letters were sent regarding 246 patients (median age 76). Fifty-three patients still had reversible ARF or died within 3 months of the letter; seven were already referred. The requesting clinician felt that referral was not appropriate in 56; 23 were being reviewed elsewhere. The programme has led to the referral of 50 patients to the renal service (and 3 to others) but in 54 cases no reply was received and the letter ignored. Of the 50 referred, 17 entered the dialysis programme, 13 of whom had definitive dialysis access (fistula or Tenckhoff catheter) at the start. After a period of outpatient review they have undergone a median of 21 months of dialysis. CONCLUSIONS: Over the 5-year period this programme has detected a cohort of patients who, in general, benefited from nephrological follow-up and dialysis. It may also act as a prompt to clinicians to refer more 'marginal' patients and thereby hasten future referral. PMID- 18390893 TI - Distributed model of peritoneal transport: implications of the endothelial glycocalyx. PMID- 18390894 TI - Synovial fluid-derived mesenchymal stem cells increase after intra-articular ligament injury in humans. AB - OBJECTIVE: The existence of mesenchymal stem cells (MSCs) in SF was previously reported. However, the behaviour and properties of MSCs derived from SF have not been fully elucidated. METHODS: Human SFs were obtained from 19 knee joints with anterior cruciate ligament injury around the time of reconstruction surgery, and from three healthy volunteers. SF was plated, cultured and examined for colony forming number, in vitro differentiation, surface epitopes and gene profiles. Also, rabbit synovium-MSCs were injected into the knee joint in a rabbit partial anterior cruciate ligament defect model, and the injected cells were traced. RESULTS: SF-MSCs from IA ligament injury patients were 100 times more in number than those from healthy volunteers. Total colony number was positively correlated with post-injury period. No significant differences were observed among the cells derived from SF around the time of the surgery in relation to surface epitopes and differentiation potentials. Cluster analysis of gene profiles demonstrated that SF-MSCs were more similar to synovium MSCs than bone marrow MSCs. In rabbit experiments, the MSCs injected into the knee in which IA ligament was partially defective were observed more on the defected area than on the intact area of the ligament at 24 h. CONCLUSION: We demonstrated that SF-MSCs, similar to synovium MSCs, increased in number after IA ligament injury and surgery without marked alteration of the properties. PMID- 18390895 TI - Haemoglobin A1c, fasting glucose, serum C-peptide and insulin resistance in relation to serum uric acid levels--the Third National Health and Nutrition Examination Survey. AB - OBJECTIVE: To evaluate haemoglobin A1c (HbA1c), fasting glucose, serum C-peptide and insulin resistance in relation to serum uric acid levels in a nationally representative sample of men and women. METHODS: Using data from 14,664 participants aged 20 yrs and older in The US Third National Health and Nutrition Examination Survey (1988-1994), we examined the relation between the levels of HbA1c, other biomarkers and serum uric acid levels using multivariate linear regressions stratified by gender. RESULTS: The serum uric acid levels increased with increasing serum HbA1c levels up to the category of 6-6.9%, and thereafter decreased with further increasing HbA1c levels (a bell-shaped relation). Compared with a HbA1c level of < 5%, the multivariate differences among women were 26.8 micromol/l for HbA1c of 6-6.9% and -25.6 micromol/l (95% CI -42.8, -8.3) for HbA1c > or = 9%. The corresponding multivariate differences among men were 8.3 micromol/l (95% CI -3.0, 19.6) and -64.8 micromol/l (95% CI -46.0, -84.5), which were both significantly different from those among women (P-values for interaction by sex <0.001). Fasting glucose levels also showed a bell-shaped relation with serum uric acid levels. Individuals with diabetes showed lower serum uric acid levels and the association was larger among men (P-value for interaction, 0.007). Serum uric acid levels increased linearly with increasing fasting serum C-peptide levels, serum insulin levels or insulin resistance (multivariate P-values for trend, <0.001). CONCLUSIONS: Individuals with moderately elevated HbA1c levels (i.e. pre-diabetes) may be at a higher risk of hyperuricaemia and gout, particularly in women, whereas individuals with diabetes or highly elevated HbA1c levels may be at a lower risk of these conditions, particularly in men. PMID- 18390896 TI - Social functioning and facial expression recognition in survivors of pediatric brain tumors. AB - OBJECTIVE: To assess social functioning and facial expression recognition skill in survivors of pediatric brain tumors (BT) as compared to children with juvenile rheumatoid arthritis (JRA). METHODS: The social functioning of 51 survivors of BT and 31 children with JRA was assessed using a facial expression recognition task, questionnaire ratings of social functioning, and an IQ screener. RESULTS: After controlling for estimated IQ, survivors of BT made significantly more errors interpreting adult facial expressions as compared to children with JRA. Additionally, history of therapy and diagnosis age predicted performance on the child portion of the facial recognition task. Finally, survivors of BT demonstrated significantly impaired social functioning across multiple measures when compared to children with JRA. CONCLUSIONS: Survivors of pediatric BT showed significant deficits in social functioning as compared to an illness comparison group. Errors in facial expression recognition represent another method for evaluating deficits that contribute to social outcomes. PMID- 18390897 TI - Insulin signalling in the heart. AB - The main role of insulin in the heart under physiological conditions is obviously the regulation of substrate utilization. Indeed, insulin promotes glucose uptake and its utilization via glycolysis. In addition, insulin participates in the regulation of long-chain fatty acid uptake, protein synthesis, and vascular tonicity. Significant advancements have been made over the last 20 years in the understanding of the signal transduction elements involved in these insulin effects. Among these molecular mechanisms, the phosphatidylinositol 3 kinase/protein kinase B (Akt) pathway is thought to play a crucial role. Under pathological conditions, such as type-2 diabetes, myocardial ischaemia, and cardiac hypertrophy, insulin signal transduction pathways and action are clearly modified. These molecular signalling alterations are often linked to atypical crosstalks with other signal transduction pathways. On the other hand, pharmacological modifications of parallel and interdependent signalling components, such as the AMP-activated protein kinase pathway, are now considered to be a good therapeutic approach to treat insulin-signalling defects such as insulin resistance and type-2 diabetes. In this review, we will focus on the description of the molecular signalling elements involved in insulin action in the heart and vasculature under these different physiological, pathological, and therapeutical conditions. PMID- 18390898 TI - Four and a half LIM protein 1: a novel chaperone for atrium-specific Kv1.5 channels with a potential role in atrial arrhythmogenesis. PMID- 18390899 TI - Uraemic hyperparathyroidism causes a reversible inflammatory process of aortic valve calcification in rats. AB - AIMS: Renal failure is associated with aortic valve calcification (AVC). Our aim was to develop an animal model for exploring the pathophysiology and reversibility of AVC, utilizing rats with diet-induced kidney disease. METHODS AND RESULTS: Sprague-Dawley rats (n = 23) were fed a phosphate-enriched, uraemia inducing diet for 7 weeks followed by a normal diet for 2 weeks ('diet group'). These rats were compared with normal controls (n = 10) and with uraemic controls fed with phosphate-depleted diet ('low-phosphate group', n = 10). Clinical investigations included serum creatinine, phosphate and parathyroid hormone (PTH) levels, echocardiography, and multislice computed tomography. Pathological examinations of the valves included histological characterization, Von Kossa staining, and antigen and gene expression analyses. Eight diet group rats were further assessed for reversibility of valve calcification following normalization of their kidney function. At 4 weeks, all diet group rats developed renal failure and hyperparathyroidism. At week 9, renal failure resolved with improvement in the hyperparathyroid state. Echocardiography demonstrated valve calcifications only in diet group rats. Tomographic calcium scores were significantly higher in the diet group compared with controls. Von Kossa stain in diet group valves revealed calcium deposits, positive staining for osteopontin, and CD68. Gene expression analyses revealed overexpression of osteoblast genes and nuclear factor kappaB activation. Valve calcification resolved after diet cessation in parallel with normalization of PTH levels. Resolution was associated with down regulation of inflammation and osteoblastic features. Low-phosphate group rats developed kidney dysfunction similar to that of the diet group but with normal levels of PTH. Calcium scores and histology showed only minimal valve calcification. CONCLUSION: We developed an animal model for AVC. The process is related to disturbed mineral metabolism. It is associated with inflammation and osteoblastic features. Furthermore, the process is reversible upon normalization of the mineral homeostasis. Thus, our model constitutes a convenient platform for studying AVC and potential remedies. PMID- 18390900 TI - IKs response to protein kinase A-dependent KCNQ1 phosphorylation requires direct interaction with microtubules. AB - AIMS: KCNQ1 (alias KvLQT1 or Kv7.1) and KCNE1 (alias IsK or minK) co-assemble to form the voltage-activated K(+) channel responsible for I(Ks)-a major repolarizing current in the human heart-and their dysfunction promotes cardiac arrhythmias. The channel is a component of larger macromolecular complexes containing known and undefined regulatory proteins. Thus, identification of proteins that modulate its biosynthesis, localization, activity, and/or degradation is of great interest from both a physiological and pathological point of view. METHODS AND RESULTS: Using a yeast two-hybrid screening, we detected a direct interaction between beta-tubulin and the KCNQ1 N-terminus. The interaction was confirmed by co-immunoprecipitation of beta-tubulin and KCNQ1 in transfected COS-7 cells and in guinea pig cardiomyocytes. Using immunocytochemistry, we also found that they co-localized in cardiomyocytes. We tested the effects of microtubule-disrupting and -stabilizing agents (colchicine and taxol, respectively) on the KCNQ1-KCNE1 channel activity in COS-7 cells by means of the permeabilized-patch configuration of the patch-clamp technique. None of these agents altered I(Ks). In addition, colchicine did not modify the current response to osmotic challenge. On the other hand, the I(Ks) response to protein kinase A (PKA)-mediated stimulation depended on microtubule polymerization in COS-7 cells and in cardiomyocytes. Strikingly, KCNQ1 channel and Yotiao phosphorylation by PKA-detected by phospho-specific antibodies-was maintained, as was the association of the two partners. CONCLUSION: We propose that the KCNQ1-KCNE1 channel directly interacts with microtubules and that this interaction plays a major role in coupling PKA-dependent phosphorylation of KCNQ1 with I(Ks) activation. PMID- 18390902 TI - Tracking quality over time: what do pressure ulcer data show? AB - OBJECTIVE: To compare the prevalence of pressure ulcers and prevention before and after a quality improvement program; determine whether patient characteristics differed for those who did and did not develop pressure ulcers; identify pressure ulcer prevention implemented at admission and whether prevention and risk factors varied by pressure ulcer severity. DESIGN: Descriptive comparative study based on two cross-sectional pressure ulcer surveys conducted in 2002 and 2006, complemented with a retrospective audit of the electronic health record and administrative system for patients identified with pressure ulcers. SETTING: 1100 bed Swedish university hospital. PARTICIPANTS: 612 hospitalized patients in 2002 and 632 in 2006. MAIN OUTCOME MEASURES: Prevalence of pressure ulcers and prevention (pressure-reducing mattresses; planned repositioning; chair, heel and 30 degrees lateral positioning cushions). RESULTS: Pressure ulcer prevalence was 23.9% in 2002 and 22.9% in 2006. When non-blanchable erythema was excluded, the prevalence was 8.0 and 12.0%, respectively. The use of pressure-reducing mattresses increased while planned repositioning decreased. Those who developed ulcers were older, at-risk for ulcers, incontinent and had longer length of stay. Little prevention was documented at admission. Some prevention strategies and risk factors were related to severity of ulcers. CONCLUSIONS: Pressure ulcer prevalence did not decrease, despite a comprehensive quality improvement program. Special attention is needed to provide prevention to older patients with acute admission. Skin and risk assessment, as well as prevention, should start early in the hospitalization. Identifying those persons with community-acquired versus hospital-acquired ulcers will strengthen pressure ulcers as an accurate marker of quality of care for hospitalized patients. If possible, data should be reported by ward level for comparison over time. PMID- 18390901 TI - AMP-activated protein kinase is activated as a consequence of lipolysis in the adipocyte: potential mechanism and physiological relevance. AB - AMP-activated protein kinase (AMPK) is activated in adipocytes during exercise and other states in which lipolysis is stimulated. However, the mechanism(s) responsible for this effect and its physiological relevance are unclear. To examine these questions, 3T3-L1 adipocytes were treated with cAMP-inducing agents (isoproterenol, forskolin, and isobutylmethylxanthine), which stimulate lipolysis and activate AMPK. When lipolysis was partially inhibited with the general lipase inhibitor orlistat, AMPK activation by these agents was also partially reduced, but the increases in cAMP levels and cAMP-dependent protein kinase (PKA) activity were unaffected. Likewise, small hairpin RNA-mediated silencing of adipose tissue triglyceride lipase inhibited both forskolin-stimulated lipolysis and AMPK activation but not that of PKA. Forskolin treatment increased the AMP:ATP ratio, and this too was reduced by orlistat. When acyl-CoA synthetase, which catalyzes the conversion of fatty acids to fatty acyl-CoA, was inhibited with triacsin C, the increases in both AMPK activity and AMP:ATP ratio were blunted. Isoproterenol stimulated lipolysis was accompanied by an increase in oxidative stress, an effect that was quintupled in cells incubated with the AMPK inhibitor compound C. The isoproterenol-induced increase in the AMP:ATP ratio was also much greater in these cells. In conclusion, the results indicate that activation of AMPK in adipocytes by cAMP-inducing agents is a consequence of lipolysis and not of PKA activation. They suggest that AMPK activation in this setting is caused by an increase in the AMP:ATP ratio that appears to be due, at least in part, to the acylation of fatty acids. Finally, this AMPK activation appears to restrain the energy depletion and oxidative stress caused by lipolysis. PMID- 18390903 TI - Mapping the functional interaction of Sco1 and Cox2 in cytochrome oxidase biogenesis. AB - Sco1 is implicated in the copper metallation of the Cu(A) site in Cox2 of cytochrome oxidase. The structure of Sco1 in the metallated and apo-conformers revealed structural dynamics primarily in an exposed region designated loop 8. The structural dynamics of loop 8 in Sco1 suggests it may be an interface for interactions with Cox17, the Cu(I) donor and/or Cox2. A series of conserved residues in the sequence motif (217)KKYRVYF(223) on the leading edge of this loop are shown presently to be important for yeast Sco1 function. Cells harboring Y219D, R220D, V221D, and Y222D mutant Sco1 proteins failed to restore respiratory growth or cytochrome oxidase activity in sco1Delta cells. The mutant proteins are stably expressed and are competent to bind Cu(I) and Cu(II) normally. Specific Cu(I) transfer from Cox17 to the mutant apo-Sco1 proteins proceeds normally. In contrast, using two in vivo assays that permit monitoring of the transient Sco1 Cox2 interaction, the mutant Sco1 molecules appear compromised in a function with Cox2. The mutants failed to suppress the respiratory defect of cox17-1 cells unlike wild-type SCO1. In addition, the mutants failed to suppress the hydrogen peroxide sensitivity of sco1Delta cells. These studies implicate different surfaces on Sco1 for interaction or function with Cox17 and Cox2. PMID- 18390904 TI - Cleaved serpin refolds into the relaxed state via a stressed conformer. AB - Serine proteinase inhibitors (serpins) are believed to fold in vivo into a metastable "stressed" state with cleavage of their P1-P1' bond resulting in reactive center loop insertion and a thermostable "relaxed" state. To understand this unique folding mechanism, we investigated the refolding processes of the P1 P1'-cleaved forms of wild type ovalbumin (cl-OVA) and the R339T mutant (cl R339T). In the native conditions, cl-OVA is trapped as the stressed conformer, whereas cl-R339T attains the relaxed structure. Under urea denaturing conditions, these cleaved proteins completely dissociated into the heavy (Gly(1)-Ala(352)) and light (Ser(353)-Pro(385)) chains. Upon refolding, the heavy chains of both proteins formed essentially the same initial burst refolding intermediates and then reassociated with the light chain counterparts. The reassociated intermediates both refolded into the native states with indistinguishable kinetics. The two refolded proteins, however, had a notable difference in thermostability. cl-OVA refolded into the stressed form with T(m) = 68.4 degrees C, whereas cl-R339T refolded into the relaxed form with T(m) = 85.5 degrees C. To determine whether cl-R339T refolds directly to the relaxed state or through the stressed state, conformational analyses by anion-exchange chromatography and fluorescence measurements were executed. The results showed that cl-R339T refolds first to the stressed conformation and then undergoes the loop insertion. This is the first demonstration that the P1-P1'-cleaved serpin peptide capable of loop insertion refolds to the stressed conformation. This highlights that the stressed conformation of serpins is an inevitable intermediate state on the folding pathway to the relaxed structure. PMID- 18390905 TI - LOX-1 receptor blockade abrogates oxLDL-induced oxidative DNA damage and prevents activation of the transcriptional repressor Oct-1 in human coronary arterial endothelium. AB - Activation of the lectin-like oxLDL receptor (LOX-1) promotes atherosclerosis. Oxidized LDL (oxLDL) increases production of reactive oxygen species (ROS) and leads to the development of endothelial dysfunction. The molecular causes for oxLDL to induce oxidative DNA damage and metabolic dysfunction remain uncertain. Here we report treatment of cultured human coronary arterial endothelial cells (HCAEC) with oxLDL to cause oxidative DNA damage as determined by a 3-fold increase in 8-OH-desoxyguanosine adduct formation and a 4-fold induction of the growth arrest and DNA damage-inducible transcripts GADD45 and GADD153. Oxidative stress resulted in activation of Oct-1, a transcriptional repressor of various vascular cytochrome P450 (CYP) monooxygenases. Activation of Oct-1 was protein kinase C (PKC)-mediated. Binding of Oct-1 to promoter sequences of CYP monooxygenases was increased upon treatment of HCAEC with oxLDL. This resulted in repressed production of endothelium-derived hyperpolarization factor 11,12 epoxyeicosatrieonic acid. Small interference RNA-mediated functional knockdown of Oct-1 prevented oxLDL-mediated silencing of CYP expression. Inhibition of LOX-1 attenuated oxLDL-mediated endothelial DNA damage, Oct-1/DNA binding, and reversed impaired production of EDHF. Taken collectively, oxLDL induced oxidative DNA damage and activation of Oct-1 to result in metabolic dysfunction of coronary arterial endothelium. PMID- 18390906 TI - Inhibition of recombinant human T-type calcium channels by Delta9 tetrahydrocannabinol and cannabidiol. AB - Delta(9)-Tetrahydrocannabinol (THC) and cannabidiol (CBD) are the most prevalent biologically active constituents of Cannabis sativa. THC is the prototypic cannabinoid CB1 receptor agonist and is psychoactive and analgesic. CBD is also analgesic, but it is not a CB1 receptor agonist. Low voltage-activated T-type calcium channels, encoded by the Ca(V)3 gene family, regulate the excitability of many cells, including neurons involved in nociceptive processing. We examined the effects of THC and CBD on human Ca(V)3 channels stably expressed in human embryonic kidney 293 cells and T-type channels in mouse sensory neurons using whole-cell, patch clamp recordings. At moderately hyperpolarized potentials, THC and CBD inhibited peak Ca(V)3.1 and Ca(V)3.2 currents with IC(50) values of approximately 1 mum but were less potent on Ca(V)3.3 channels. THC and CBD inhibited sensory neuron T-type channels by about 45% at 1 mum. However, in recordings made from a holding potential of -70 mV, 100 nm THC or CBD inhibited more than 50% of the peak Ca(V)3.1 current. THC and CBD produced a significant hyperpolarizing shift in the steady state inactivation potentials for each of the Ca(V)3 channels, which accounts for inhibition of channel currents. Additionally, THC caused a modest hyperpolarizing shift in the activation of Ca(V)3.1 and Ca(V)3.2. THC but not CBD slowed Ca(V)3.1 and Ca(V)3.2 deactivation and inactivation kinetics. Thus, THC and CBD inhibit Ca(V)3 channels at pharmacologically relevant concentrations. However, THC, but not CBD, may also increase the amount of calcium entry following T-type channel activation by stabilizing open states of the channel. PMID- 18390907 TI - Hydrolysis of phosphatidylinositol 4,5-bisphosphate mediates calcium-induced inactivation of TRPV6 channels. AB - TRPV6 is a member of the transient receptor potential superfamily of ion channels that facilitates Ca(2+) absorption in the intestines. These channels display high selectivity for Ca(2+), but in the absence of divalent cations they also conduct monovalent ions. TRPV6 channels have been shown to be inactivated by increased cytoplasmic Ca(2+) concentrations. Here we studied the mechanism of this Ca(2+) induced inactivation. Monovalent currents through TRPV6 substantially decreased after a 40-s application of Ca(2+), but not Ba(2+). We also show that Ca(2+), but not Ba(2+), influx via TRPV6 induces depletion of phosphatidylinositol 4,5 bisphosphate (PI(4,5)P(2) or PIP(2)) and the formation of inositol 1,4,5 trisphosphate. Dialysis of DiC(8) PI(4,5)P(2) through the patch pipette inhibited Ca(2+)-dependent inactivation of TRPV6 currents in whole-cell patch clamp experiments. PI(4,5)P(2) also activated TRPV6 currents in excised patches. PI(4)P, the precursor of PI(4,5)P(2), neither activated TRPV6 in excised patches nor had any effect on Ca(2+)-induced inactivation in whole-cell experiments. Conversion of PI(4,5)P(2) to PI(4)P by a rapamycin-inducible PI(4,5)P(2) 5 phosphatase inhibited TRPV6 currents in whole-cell experiments. Inhibiting phosphatidylinositol 4 kinases with wortmannin decreased TRPV6 currents and Ca(2+) entry into TRPV6-expressing cells. We propose that Ca(2+) influx through TRPV6 activates phospholipase C and the resulting depletion of PI(4,5)P(2) contributes to the inactivation of TRPV6. PMID- 18390908 TI - The mechanism of assembly and cofactor insertion into Rhodobacter capsulatus xanthine dehydrogenase. AB - Rhodobacter capsulatus xanthine dehydrogenase (XDH) is a molybdo-flavoprotein that is highly homologous to the homodimeric mammalian xanthine oxidoreductase. However, the bacterial enzyme has an (alphabeta)(2) heterotetrameric structure, and the cofactors were identified to be located on two different polypeptides. We have analyzed the mechanism of cofactor insertion and subunit assembly of R. capsulatus XDH, using engineered subunits with appropriate substitutions in the interfaces. In an (alphabeta) heterodimeric XDH containing the XdhA and XdhB subunits, the molybdenum cofactor (Moco) was shown to be absent, indicating that dimerization of the (alphabeta) subunits has to precede Moco insertion. In an (alphabeta)(2) XDH heterotetramer variant, including only one active Moco-center, the active (alphabeta) site of the chimeric enzyme was shown to be fully active, revealing that the two subunits act independent without cooperativity. Amino acid substitutions at two cysteine residues coordinating FeSI of the two [2Fe-2S] clusters of the enzyme demonstrate that an incomplete assembly of FeSI impairs the formation of the XDH (alphabeta)(2) heterotetramer and, thus, insertion of Moco into the enzyme. The results reveal that the insertion of the different redox centers into R. capsulatus XDH takes place sequentially. Dimerization of two (alphabeta) dimers is necessary for insertion of sulfurated Moco into apo XDH, the last step of XDH maturation. PMID- 18390909 TI - Validity of enthesis ultrasound assessment in spondyloarthropathy. AB - OBJECTIVES: To develop an ultrasound enthesis score and to assess its validity in the diagnostic classification of the spondyloarthropathies (SpAs). METHODS: Twenty-five patients with SpA and 29 healthy controls participated in a blinded, gender-matched, cross-sectional study involving ultrasound assessment. The following entheses were explored bilaterally: proximal plantar fascia, distal Achilles tendon, distal and proximal patellar ligament, distal quadriceps and brachial triceps tendons. The ultrasound score evaluated enthesis thickness, structure, calcifications, erosions, bursae and power Doppler signal. The value of each elemental lesion was calculated using a three-model analysis. Validity was analysed by receiver operating characteristic (ROC) curves. Inter-reader and interexplorer intraclass correlation coefficients (ICCs) were calculated. RESULTS: The logistic regression model overestimated the score of three elemental lesions: calcification (0-3), Doppler (0 or 3) and erosion (0 or 3), while scoring tendon structure, tendon thickness and bursa as 0 or 1. ROC curves established an ultrasound score of >or=18 as the best cut-off point for differentiation between cases and controls. This cut-off point was exceeded by 5/29 controls (17%) and by 21/25 patients with SpA (84%). The sensitivity, specificity, positive and negative likelihood ratios (LR+, LR-) were 83.3%, 82.8%, 4.8% and 0.2%, respectively. The inter-reader and interexplorer ICCs were 0.60 and 0.86, respectively. CONCLUSION: The findings suggest that the ultrasound enthesis score could be a valid tool in the diagnosis of SpA. PMID- 18390910 TI - Quantifying anti-cyclic citrullinated peptide titres: clinical utility and association with tobacco exposure in patients with rheumatoid arthritis. AB - OBJECTIVE: To determine the significance of quantitative levels of antibodies to cyclic citrullinated peptides (anti-CCP) in a population of patients with rheumatoid arthritis (RA). METHODS: A total of 241 consecutive sera from patients with RA sent from a large rheumatology clinic for laboratory testing were selected for precisely quantifying anti-CCP antibody titres with the anti-CCP2 assay. Patient charts were reviewed for demographic information, smoking history, clinical diagnosis, rheumatoid factor (RF) titre, radiographic information and other laboratory information (erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) level). Correlations with anti-CCP titre and RF titre, disease parameters and smoking history were assessed. RESULTS: We confirm previous findings that anti-CCP seropositivity is associated with a higher incidence of erosions in patients with RA (56% vs 20% CCP+ vs CCP-, kappa = 0.297, p<0.001). We also found a moderate correlation between anti-CCP titre and RF titre. However, we failed to find an association between anti-CCP titre and presence of erosions, between anti-CCP titre and CRP or ESR level, or between anti-CCP titre and age or disease duration. Interestingly, we did find significantly higher anti CCP titres in patients with a history of smoking (452 units/ml vs 229 units/ml, smokers vs non-smokers, respectively; p = 0.02). CONCLUSIONS: Although anti-CCP titres were not associated with clinical parameters of disease, they are increased in patients with RA with exposure to tobacco. By contrast, no elevation in RF was noted in patients with a history of smoking. These observations are consistent with a pathogenic contribution of smoking to RA and suggest the immune stimulus for anti-CCP is distinct from that for RF. PMID- 18390911 TI - Consequences of dramatic reductions in state tobacco control funds: Florida, 1998 2000. AB - OBJECTIVE: This study assessed whether dramatic funding reductions to the Florida Tobacco Control Program (FTCP) influenced trends in recall of the Florida "truth" anti-smoking media campaign, anti-industry attitudes and non-smoking intentions among Florida teens. METHODS: We used an interrupted time series technique to test for differences in the rates of change in Florida "truth" recall, anti industry beliefs and non-smoking intentions before and after the FTCP budget cuts using the Florida Anti-tobacco Media Evaluation (FAME) survey, a repeated cross sectional telephone survey of Florida teens. RESULTS: Recall of the Florida "truth" anti-smoking campaign, anti-industry attitudes, and non-smoking intentions increased dramatically between April 1998 and May 1999. Florida "truth" recall declined after FTCP budget cuts in June 1999. Anti-industry beliefs and non-smoking intentions plateaued or began to decline after the budget cuts. The launch of the national "truth" campaign in February 2000 may have offset otherwise deleterious effects of the budget cuts on anti-industry beliefs, but not smoking intentions. CONCLUSION: Reductions in tobacco control funding have immediate effects on programme exposure and cognitive precursors to smoking initiation. There is a critical need to maintain and enhance funding for state tobacco control programmes to continue nationwide progress in preventing youth from initiating cigarette smoking. PMID- 18390912 TI - Cardiopulmonary resuscitation for out of hospital cardiac arrest. PMID- 18390913 TI - Surgical treatment of pilonidal disease. PMID- 18390914 TI - Healing by primary closure versus open healing after surgery for pilonidal sinus: systematic review and meta-analysis. AB - OBJECTIVE: To determine the relative effects of open healing compared with primary closure for pilonidal sinus and optimal closure method (midline v off midline). DESIGN: Systematic review and meta-analyses of randomised controlled trials. DATA SOURCES: Cochrane register of controlled trials, Cochrane Wounds Group specialised trials register, Medline (1950-2007), Embase, and CINAHL bibliographic databases, without language restrictions. DATA EXTRACTION: Primary outcomes were time (days) to healing, surgical site infection, and recurrence rate. Secondary outcomes were time to return to work, other complications and morbidity, cost, length of hospital stay, and wound healing rate. STUDY SELECTION: Randomised controlled trials evaluating surgical treatment of pilonidal sinus in patients aged 14 years or more. Data were extracted independently by two reviewers and assessed for quality. Meta-analyses used fixed and random effects models, dichotomous data were reported as relative risks or Peto odds ratios and continuous data are given as mean differences; all with 95% confidence intervals. RESULTS: 18 trials (n=1573) were included. 12 trials compared open healing with primary closure. Time to healing was quicker after primary closure although data were unsuitable for aggregation. Rates of surgical site infection did not differ; recurrence was less likely to occur after open healing (relative risk 0.42, 0.26 to 0.66). 14 patients would require their wound to heal by open healing to prevent one recurrence. Six trials compared surgical closure methods (midline v off-midline). Wounds took longer to heal after midline closure than after off-midline closure (mean difference 5.4 days, 95% confidence interval 2.3 to 8.5), rate of infection was higher (relative risk 4.70, 95% confidence interval 1.93 to 11.45), and risk of recurrence higher (Peto odds ratio 4.95, 95% confidence interval 2.18 to 11.24). Nine patients would need to be treated by an off-midline procedure to prevent one surgical site infection and 11 would need to be treated to prevent one recurrence. CONCLUSIONS: Wounds heal more quickly after primary closure than after open healing but at the expense of increased risk of recurrence. Benefits were clearly shown with off-midline closure compared with midline closure. Off-midline closure should become standard management for pilonidal sinus when closure is the desired surgical option. PMID- 18390915 TI - Preventing sports injuries at the national level: time for other nations to follow New Zealand's remarkable success. PMID- 18390916 TI - Injury surveillance in multi-sport events: the International Olympic Committee approach. AB - BACKGROUND: The protection of athletes' health by preventing injuries is an important task for international sports federations. Standardised injury surveillance provides not only important epidemiological information, but also directions for injury prevention, and the opportunity for monitoring long-term changes in the frequency and circumstances of injury. Numerous studies have evaluated sports injuries during the season, but few have focused on injuries during major sport events such as World Championships, World Cups or the Olympic Games. OBJECTIVES: To provide an injury surveillance system for multi-sports tournaments, using the 2008 Olympic Games in Beijing as an example. METHODS: A group of experienced researchers reviewed existing injury report systems and developed a scientific sound and concise injury surveillance system for large multi-sport events. RESULTS: The injury report system for multi-sport events is based on an established system for team sports tournaments and has proved feasible for individual sports during the International Association of Athletics Federations World Championships in Athletics 2007. The most important principles and advantages of the system are comprehensive definition of injury, injury report by the physician responsible for the athlete, a single-page report of all injuries, and daily report irrespective of whether or not an injury occurred. Implementation of the injury surveillance system, all definitions, the report form, and the analysis of data are described in detail to enable other researchers to implement the injury surveillance system in any sports tournament. CONCLUSION: The injury surveillance system has been accepted by experienced team physicians and shown to be feasible for single-sport and multi-sport events. It can be modified depending on the specific objectives of a certain sport or research question; however, a standardised use of injury definition, report forms and methodology will ensure the comparability of results. PMID- 18390917 TI - Heart rate and exercise intensity during training: observations from the DREW Study. AB - OBJECTIVE: Cardiovascular drift (CVdrift) is characterised by a continuous, gradual increase in heart rate (HR) after approximately 10 min of moderate intensity aerobic exercise, despite maintenance of a constant work rate. This has important implications for trials that employ HR to monitor exercise intensity, as reducing work rate in order to keep HR constant could result in participants exercising below the intended intensity. Utilising the Dose Response to Exercise in Women (DREW) database, we sought to determine if increases in HR during exercise (CVdrift) resulted in clinically significant reductions in exercise work rate in order to keep HR within a target range. DESIGN: Randomised, prospective study. SETTING: DREW clinical exercise trail, The Cooper Institute, Dallas, Texas. PARTICIPANTS: Overweight (body mass index 25-43 kg/m2), previously sedentary postmenopausal women (n=326). INTERVENTION: Treadmill and cycling exercise (30-90 min, three to five times per week) at a HR corresponding to 50% of peak oxygen uptake (VO2peak). MAIN OUTCOME MEASURE: Changes in exercise intensity (metabolic equivalents (METS)) during exercise in response to CVdrift. RESULTS: We observed small increases in HR (1-4 beats per minute, p<0.001) combined with small increases in intensity (0.01-0.03 METS, p<0.03) during the combined 12 963 exercise training sessions. Further, we identified only 101 (0.78%) sessions in which intensity was reduced during the course of the exercise session, potentially in response to CVdrift. CONCLUSIONS: We conclude that CVdrift did not contribute to significant reductions in exercise intensity in the DREW study. PMID- 18390918 TI - How did A V Hill understand the VO2max and the "plateau phenomenon"? Still no clarity? AB - OBJECTIVES: A recent editorial in Medicine and Science in Sports and Exercise concludes that the study of Hawkins and colleagues confirms "beyond any doubt the proposition of Hill and Lupton". It is not clear which of Hill and Lupton's propositions have been proved "beyond any doubt". METHODS: A review of all the relevant publications of A V Hill and his colleagues. RESULTS: Hill and Lupton believed (i) that myocardial ischaemia limits maximal exercise performance by inducing circulatory failure; (ii) that a "governor" protects the ischaemic heart by causing a "slowing of the circulation" during maximal exercise; (iii) that the oxygen cost of running increases exponentially at speeds above 16 km per hour; and (iv) that humans reach their highest measurable oxygen consumption of about 4 litres per minute at that running speed. Hill and Lupton neither invented the concept of the "plateau phenomenon" nor utilised this concept to establish that a "true" maximum oxygen consumption (VO2max) had been achieved. Nor did they measure cardiac output during exercise. CONCLUSION: Accepting uncritically this modern interpretation of Hill and Lupton's theory that the cardiac output limits maximal exercise performance whether or not the plateau phenomenon is present fails to answer the question first posed more than 20 years ago: What causes the termination of exercise when the "true" VO2max is achieved without the "plateau phenomenon"? According to the Hill and Lupton model, this cannot be because a limiting cardiac output has been reached. Since a majority of maximal exercise tests terminate in the absence of the "plateau phenomenon", this is seemingly an important question. PMID- 18390919 TI - Effect of proprioception training on knee joint position sense in female team handball players. AB - BACKGROUND: A number of studies have shown that proprioception training can reduce the risk of injuries in pivoting sports, but the mechanism is not clearly understood. AIM: To determine the contributing effects of propioception on knee joint position sense among team handball players. STUDY DESIGN: Prospective cohort study. METHODS: Two professional female handball teams were followed prospectively for the 2005-6 season. 20 players in the intervention team followed a prescribed proprioceptive training programme while 19 players in the control team did not have a specific propioceptive training programme. The coaches recorded all exposures of the individual players. The location and nature of injuries were recorded. Joint position sense (JPS) was measured by a goniometer on both knees in three angle intervals, testing each angle five times. Assessments were performed before and after the season by the same examiner for both teams. In the intervention team a third assessment was also performed during the season. Complete data were obtained for 15 subjects in the intervention team and 16 in the control team. Absolute error score, error of variation score and SEM were calculated and the results of the intervention and control teams were compared. RESULTS: The proprioception sensory function of the players in the intervention team was significantly improved between the assessments made at the start and the end of the season (mean (SD) absolute error 9.78-8.21 degrees (7.19 6.08 degrees ) vs 3.61-4.04 degrees (3.71-3.20 degrees ), p<0.05). No improvement was seen in the sensory function in the control team between the start and the end of the season (mean (SD) absolute error 6.31-6.22 degrees (6.12-3.59 degrees ) vs 6.13-6.69 degrees (7.46-6.49 degrees ), p>0.05). CONCLUSION: This is the first study to show that proprioception training improves the joint position sense in elite female handball players. This may explain the effect of neuromuscular training in reducing the injury rate. PMID- 18390920 TI - The risk for a cruciate ligament injury of the knee in adolescents and young adults: a population-based cohort study of 46 500 people with a 9 year follow-up. AB - OBJECTIVES: The knee joint is the most common site for injury among younger people, the injury often resulting in expensive (surgical) treatment, long-term rehabilitation and permanent functional impairment and disability. METHODS: We investigated the incidence and risk factors for a major knee-ligament injury in an adolescent Finnish population. A population-based cohort of 46 472 adolescents was followed for an average of 9 years. All patients hospitalised with the diagnosis of anterior or posterior cruciate ligament injury (ACL or PCL injury) were included in the analysis. RESULTS: 265 (0.6%) people (194 male and 71 female subjects) from the total cohort of 46 472 were treated for a cruciate ligament injury of the knee during the follow-up period, giving an injury incidence of 60.9 (95% CI 53.6 to 68.2) per 100 000 person-years. When the socioeconomic, health and lifestyle background variables were taken into account, the adjusted hazard ratio for a cruciate ligament injury of the knee was 8.5 (95% CI 4.3 to 16.4) for female and 4.0 (95% CI 2.7 to 6.1) for male subjects who participated in organised sports > or =4 times/week. CONCLUSIONS: The general risk for a cruciate ligament injury of the knee is relatively low among adolescents and young adults, but participation in organised sports increases the risk significantly. The risk is especially high in active young women. Preventive measures should be adopted to decrease the short-term and long-term burden of these severe injuries. PMID- 18390921 TI - Interim evaluation of the effect of a new scrum law on neck and back injuries in rugby union. AB - BACKGROUND: In January 2007 the International Rugby Board implemented a new law for scrum engagement aimed at improving player welfare by reducing impact force and scrum collapses. In New Zealand the new law was included in RugbySmart, an annual compulsory workshop for coaches and referees. OBJECTIVE: To determine the effect of the new law on scrum-related moderate to serious neck and back injury claims in 2007. METHODS: Claims filed with the Accident Compensation Corporation (the provider of no-fault injury compensation and rehabilitation in New Zealand) were combined with numbers of registered players to estimate moderate to serious scrum-related claims for players who take part in scrums (forwards). Poisson linear regression was used to compare the observed claims per 100 000 forwards for 2007 with the rate predicted from data for 2002-6. RESULTS: The observed and predicted claims per 100 000 forwards were 52 and 76, respectively (rate ratio 0.69; 90% CI 0.42 to 1.12). The likelihoods of substantial benefit (rate ratio <0.90) and harm (rate ratio >1.1) attributable to the scrum law were 82% and 5%, respectively. CONCLUSION: The decline in scrum-related injury claims is consistent with a beneficial effect of the new scrum law in the first year of its implementation. Another year of monitoring should provide more evidence for the efficacy of the new law. PMID- 18390922 TI - Use of field-based tests to identify risk factors for injury to fast bowlers in cricket. AB - OBJECTIVE: To identify risk factors for injury to cricket fast bowlers using field-based tests. DESIGN: Prospective cohort study. SETTING: High performance Australian cricket. PARTICIPANTS: Ninety-one male adolescent and adult fast bowlers (aged 12-33 years). ASSESSMENT OF RISK FACTORS: A field-based pre participation screening, consisting of musculoskeletal, fitness and anthropometric assessments and analysis of bowling technique was undertaken. Bowlers were prospectively monitored over the 2003-4 season and bowling workload and injuries were recorded. Logistic regression was used to identify injury risk factors. MAIN OUTCOME MEASUREMENT: Repetitive microtrauma injury to the trunk, back or lower limb associated with fast bowling. RESULTS: Two variables were identified as independent predictors of injury in the multivariate logistic regression analysis. Bowlers with hip internal rotation of < or =30 degrees on the leg ipsilateral to the bowling arm were at a significantly reduced risk of injury (OR 0.20, 95% CI 0.06 to 0.73) compared with bowlers with >40 degrees of rotation. Bowlers with an ankle dorsiflexion lunge of 12.1-14.0 cm on the leg contralateral to the bowling arm were at a significantly increased risk (OR 4.03, 95% CI 1.07 to 15.21) than bowlers with a lunge of >14 cm. Bowlers with a lunge of < or =12 cm were also at an increased risk, but not significantly so (OR 1.38, 95% CI 0.40 to 4.84). CONCLUSIONS: Biomechanical research is needed to investigate how these two intrinsic risk factors increase injury risk so that appropriate interventions can be developed. PMID- 18390923 TI - Glucogenic supply increases ovulation rate by modifying follicle recruitment and subsequent development of preovulatory follicles without effects on ghrelin secretion. AB - This study determined the effects of short-term energy inputs on ghrelin secretion and possible links with changes in the follicle population or the ovulation rate. Oestrous cycle was synchronized in 16 Manchega sheep using progestagen sponges and cloprostenol. Half of the animals were treated from days 0 to 4 by the oral administration, twice daily, of 200 ml of a glucogenic mixture containing 70% of glycerol, 20% of 1,2-propanediol and 10% of water; the control group received 200 ml water. The mean (+/-S.E.M.) plasma glucose increased immediately after the first administration (3.9+/-0.3 vs 3.0+/-0.1 mmol/l in control group, P<0.05), remaining statistically different during the treatment. However, plasma ghrelin levels were similar in both groups. On the other hand, the results indicated that short-term energy inputs modify ovulation rate (1.9+/ 0.1 vs 1.3+/-0.2 in control group, P<0.05) by increasing the number of follicles able to be selected to ovulate during the period of treatment (>or=4 mm in size; 5.9+/-0.6 vs 4.3+/-0.4 at day 2, P<0.05). After sponge withdrawal, the number of these follicles decreased throughout follicular phase (5.8+/-0.8 to 1.5+/-0.4, P<0.0005) while the number of large follicles increased (>or=6 mm in size; 0.8+/ 0.4 to 2.0+/-0.3, P<0.05); this would indicate an active growth of preovulatory follicles that were not found in the control group. Thus, the increases of ovulation rate by high-energy inputs would be caused by an enhancement in the developmental competence of preovulatory follicles. PMID- 18390924 TI - IFN-gamma enhances neurogenesis in wild-type mice and in a mouse model of Alzheimer's disease. AB - The generation of new neurons and glia from a precursor stem cell appears to take place in the adult brain. However, new neurons generated in the dentate gyrus decline sharply with age and to an even greater extent in neurodegenerative diseases. Here we raise the question whether peripheral immune mechanisms can generate immunity to such deficits in neuronal repair. We demonstrate that in contrast to primarily innate immunity cytokines, such as interleukin-6 and tumor necrosis factor-alpha, the adaptive immunity cytokine IFN-gamma enhances neurogenesis in the dentate gyrus of adult mice and improves the spatial learning and memory performance of the animals. In older mice, the effect of IFN-gamma is more pronounced in both wild-type mice and mice with Alzheimer's-like disease and is associated with neuroprotection. In addition, IFN-gamma reverses the increase in oligodendrogenesis observed in a mouse model of Alzheimer's disease. We demonstrate that limited amounts of IFN-gamma in the brain shape the neuropoietic milieu to enhance neurogenesis, possibly representing the normal function of the immune system in controlling brain inflammation and repair. PMID- 18390925 TI - Impaired skeletal muscle development and function in male, but not female, genomic androgen receptor knockout mice. AB - To identify mechanisms of anabolic androgen action in muscle, we generated male and female genomic androgen receptor (AR) knockout (ARKO) mice, and characterized muscle mass, contractile function, and gene expression. Muscle mass is decreased in ARKO males, but normal in ARKO females. The levator ani muscle, which fails to develop in normal females, is also absent in ARKO males. Force production is decreased from fast-twitch ARKO male muscle, and slow-twitch muscle has increased fatigue resistance. Microarray analysis shows up-regulation of genes encoding slow-twitch muscle contractile proteins. Real-time PCR confirms that expression of genes encoding polyamine biosynthetic enzymes, ornithine decarboxylase (Odc1), and S-adenosylmethionine decarboxylase (Amd1), is reduced in ARKO muscle, suggesting androgens act through regulation of polyamine biosynthesis. Altered expression of regulators of myoblast progression from proliferation to terminal differentiation suggests androgens also promote muscle growth by maintaining myoblasts in the proliferate state and delaying differentiation (increased Cdkn1c and Igf2, decreased Itg1bp3). A similar pattern of gene expression is observed in orchidectomized male mice, during androgen withdrawal-dependent muscle atrophy. In conclusion, androgens are not required for peak muscle mass in females. In males, androgens act through the AR to regulate multiple gene pathways that control muscle mass, strength, and fatigue resistance. PMID- 18390926 TI - The extreme C-terminal region of phospholipase Cbeta1 determines subcellular localization and function; the "b" splice variant mediates alpha1-adrenergic receptor responses in cardiomyocytes. AB - Phospholipase Cbeta1 (PLCbeta1) exists as two splice variants, PLCbeta1a (150 kDa) and PLCbeta1b (140 kDa), which differ only in their C-terminal sequences of 64 and 31 amino acids, respectively. The 3 C-terminal amino acid residues of PLCbeta1a comprise a PDZ-interacting domain, whereas the PLCbeta1b sequence has no PDZ-interacting domain but contains unique proline-rich domain 5 residues from the C terminus. PLCbeta1a is localized in the cytoplasm, whereas PLCbeta1b targets to the sarcolemma and is enriched in caveolae. Deletion of 3 amino acids from the C terminus of PLCbeta1b did not alter its sarcolemmal localization, but deletion of the entire unique 31 amino acid sequence caused cytosolic localization. A myristoylated 10 amino acid peptide from the C terminus of PLCbeta1b selectively dissociated N-terminally GFP-tagged PLCbeta1b from the sarcolemma and inhibited PLC responses to alpha(1)-adrenergic agonists, with a half maximal effective concentration of 12 +/- 1.6 microM (mean+/-SE, n=3). A similar peptide from PLCbeta1a was without effect at concentrations below 100 microM. Thus, the extreme C-terminal sequences of the PLCbeta1 splice variants determine localization and, thus, function. In cardiomyocytes, responses initiated by alpha(1)-adrenergic receptor activation involve only PLCbeta1b, and the selective targeting of this splice variant to the sarcolemma provides a potential therapeutic target to reduce hypertrophy, apoptosis, and arrhythmias. PMID- 18390927 TI - Priming of neutrophil oxidative burst in diabetes requires preassembly of the NADPH oxidase. AB - Hyperglycemia associated with diabetes mellitus results in the priming of neutrophils leading to oxidative stress that is, in part, responsible for diabetic complications. p47phox, a NADPH oxidase cytosolic subunit, is a key protein in the assembly of the NADPH oxidase leading to superoxide generation. Little is known about the priming mechanism of oxidative pathways in neutrophils of people with diabetes. In this study, the kinetics of p47phox activation was investigated by comparing neutrophils from diabetic and healthy subjects, and the mechanism of hyperglycemia-induced changes was studied by using neutrophil-like HL-60 cells as a model. In resting neutrophils from diabetic subjects, p47phox prematurely translocates to the cell membrane and preassembles with p22phox, a NADPH oxidase membrane subunit. This premature p47phox translocation and preassembly with p22phox were also observed in HL-60 cells cultured with high glucose (HG; 25 mM) and with the specific ligand for the receptor for advanced glycation end products (RAGE), S100B. Phosphorylation of ERK1/2, but not p38 MAPK, was the primary signaling pathway, as evidenced by PD98059 suppressing the translocation of p47phox in HL-60 cells incubated with HG and S100B. HL-60 cells cultured in HG and S100B exhibited a 1.8-fold increase in fMLP-induced superoxide generation compared with those cultured in normal glucose (5.5 mM). These data suggest that HG and increased AGE prime neutrophils and increase oxidative stress inducing the translocation of p47phox to the cell membrane and preassembly with p22phox by stimulating a RAGE-ERK1/2 pathway. PMID- 18390928 TI - Phlebotomine salivas inhibit immune inflammation-induced neutrophil migration via an autocrine DC-derived PGE2/IL-10 sequential pathway. AB - In the present study, we investigated whether saliva from Phlebotomus papatasi and Phlebotomus duboscqi inhibited antigen-induced neutrophil migration and the mechanisms involved in these effects. The pretreatment of immunized mice with salivary gland extracts (SGE) of both phlebotomines inhibited OVA challenge induced neutrophil migration and release of the neutrophil chemotactic mediators, MIP-1alpha, TNF-alpha, and leukotriene B4 (LTB4). Furthermore, SGE treatment enhanced the production of anti-inflammatory mediators, IL-10 and PGE2. SGE treatments failed to inhibit neutrophil migration and MIP-1alpha and LTB4 production in IL-10-/- mice, also failing in mice treated with nonselective (indomethacin) or selective (rofecoxibe) cyclooxygenase (COX) inhibitors. COX inhibition resulted in diminished SGE-induced IL-10 production, and PGE2 release triggered by SGE remained increased in IL-10-/- mice, suggesting that prostanoids are acting through an IL-10-dependent mechanism. SGE treatments in vivo reduced the OVA-induced lymphoproliferation of spleen-derived cells. Further, the in vitro incubation of bone marrow-derived dendritic cells (DC) with SGE inhibited the proliferation of CD4+T cells from OVA-immunized mice, which was reversed by indomethacin and anti-IL-10 antibody treatments. Supporting these results, SGE induced the production of PGE2 and IL-10 by DC, which were blocked by COX inhibition. These effects were associated with the reduction of DC-membrane expression of MHC-II and CD86 by SGE treatment. Altogether, the results showed that Phlebotomine saliva inhibits immune inflammation-induced neutrophil migration by an autocrine DC sequential production of PGE2/IL-10, suggesting that the saliva constituents might be promising therapeutic molecules to target immune inflammatory diseases. PMID- 18390930 TI - Tissue plasminogen activator (t-PA) promotes neutrophil degranulation and MMP-9 release. AB - Recombinant tissue plasminogen activator (t-PA), the only approved stroke treatment, is used for clot lysis within the occluded brain artery. Unfortunately, matrix metalloproteinase-9 (MMP-9) concentration increases after t PA treatment and has been related to hemorrhagic transformation after ischemic stroke. Although the exact cellular source of brain MMP-9 remains unknown, invading, inflammatory cells, such as neutrophils, release MMP-9 to cross the blood brain barrier. Therefore, we hypothesize that the most feared side effect of stroke reperfusion therapy, brain hemorrhage, is related to t-PA-induced MMP-9 release by neutrophils. We show by means of ELISA that t-PA treatment promotes MMP-9, MMP-8, and tissue inhibitor metalloproteinase-2 release from human neutrophils ex vivo within 10 and 30 min. Moreover, by zymography and Western blot, we observed that neutrophils are emptied of MMP-9 content after t-PA treatment at those times. Finally, total internal reflection fluorescent imaging allowed us to observe the t-PA effect on neutrophils, showing the promotion of degranulation on these cells in vivo. Our data suggest that neutrophils are good candidates to be the main source of MMP-9 following t-PA stroke treatment and in consequence, partially responsible for thrombolysis-related brain bleedings. PMID- 18390929 TI - Enteral glutamine: a novel mediator of PPARgamma in the postischemic gut. AB - Early enteral nutrition supplemented with glutamine, arginine, omega-3 fatty acids, and nucleotides has been shown to decrease infection complications in critically injured patients. Concern has been raised, however, that under conditions of hyperinflammation, these diets may be injurious through the induction of inducible NO synthase by enteral arginine. In a rodent model of gut ischemia/reperfusion, inflammation and injury are intensified by enteral arginine and abrogated by glutamine. These findings correlate with the degree of metabolic stress imposed upon the gut by hypoperfusion. Glutamine is metabolized by the gut and therefore, can contribute back energy in the form of ATP, whereas arginine is a nonmetabolizable nutrient, using but not contributing energy. Recent data suggest that one of the molecular mechanisms responsible for the gut-protective effects of enteral glutamine is the activation of peroxisome proliferator activated receptor gamma. This anti-inflammatory transcription factor belongs to the family of nuclear receptors, plays a key role in adipocyte development and glucose homeostasis, and has been recognized as an endogenous regulator of intestinal inflammation. Preliminary clinical studies support the use of enteral glutamine in patients with gut hypoperfusion. PMID- 18390931 TI - Investigative report: 1766 boil-water advisories now in place across Canada. PMID- 18390932 TI - Safe water? Depends on where you live! PMID- 18390933 TI - Growth generates health care challenges in booming India. PMID- 18390934 TI - Despite federal promises, First Nations' water problems persist. PMID- 18390935 TI - Private health clinics remain unregulated in most of Canada. PMID- 18390938 TI - Preparing for Afghanistan's medical future. PMID- 18390939 TI - The dietary supplement 5-hydroxytryptophan and urinary 5-hydroxyindole acetic acid. PMID- 18390940 TI - Giant calculi in urethral diverticula. PMID- 18390941 TI - Chronic contained rupture of an abdominal aortic aneurysm with vertebral erosion. PMID- 18390942 TI - Screening and case-finding instruments for depression: a meta-analysis. AB - BACKGROUND: Screening and case-finding has been proposed as a simple, quick and cheap method to improve the quality of care for depression. We sought to establish the effectiveness of screening in improving the recognition of depression, the management of depression and the outcomes of patients with depression. METHODS: We performed a Cochrane systematic review of randomized controlled trials conducted in nonmental health settings that included case finding or screening instruments for depression. We conducted a meta-analysis and explored heterogeneity using meta-regression techniques. RESULTS: Sixteen studies with 7576 patients met our inclusion criteria. We found that the use of screening or case-finding instruments were associated with a modest increase in the recognition of depression by clinicians (relative risk [RR] 1.27, 95% confidence interval [CI] 1.02 to 1.59). Questionnaires, when administered to all patients and the results given to clinicians irrespective of baseline score, had no impact on recognition (RR 1.03, 95% CI 0.85 to 1.24). Screening or case finding increased the use of any intervention by a relative risk of 1.30 (95% CI 0.97 to 1.76). There was no evidence of influence on the prescription of antidepressant medications (RR 1.20, 95% CI 0.87 to 1.66). Seven studies provided data on outcomes of depression, and no evidence of an effect was found (standardized mean difference -0.02, 95% CI -0.25 to 0.20). INTERPRETATION: If used alone, case finding or screening questionnaires for depression appear to have little or no impact on the detection and management of depression by clinicians. Recommendations to adopt screening strategies using standardized questionnaires without organizational enhancements are not justified. PMID- 18390944 TI - Examining asthma quality of care using a population-based approach. AB - BACKGROUND: Asthma accounts for considerable burden on health care, but in most cases, asthma can be controlled. Quality-of-care indicators would aid in monitoring asthma management. We describe the quality of asthma care using a set of proposed quality indicators. METHODS: We performed a retrospective cross sectional study using health databases in Saskatchewan, a Canadian province with a population of about 1 million people. We assessed 6 quality-of-care indicators among people with asthma: admission to hospital because of asthma; poor asthma control (high use of short-acting beta-agonists, admission to hospital because of asthma or death due to asthma); no inhaled corticosteroid use among patients with poor control; at least moderate inhaled corticosteroid use among patients with poor control; high inhaled corticosteroid use and use of another preventer medication among patients with poor control; and any main preventer use among patients with poor control. We calculated crude and adjusted rates with 95% confidence intervals. We tested for differences using the chi2 test for proportions and generalized linear modelling techniques. RESULTS: In 2002/03, there were 24 616 people aged 5-54 years with asthma in Saskatchewan, representing a prevalence of 3.8%. Poor symptom control was observed in 18% of patients with asthma. Among those with poor control, 37% were not dispensed any inhaled corticosteroids, and 40% received potentially inadequate doses. Among those with poor control who were dispensed high doses of inhaled corticosteroids, 26% also used another preventer medication. Hospital admissions because of asthma were highest among those aged 6-9 years and females aged 20-44 years. Males and those in adult age groups (predominantly 20-44 years) had worse quality of care for 4 indicators examined. INTERPRETATION: Suboptimal asthma management would be improved through increased use of inhaled corticosteroids and preventer medications, and reduced reliance on short-acting beta-agonist medications as recommended by consensus guidelines. PMID- 18390943 TI - Effect of regulatory warnings on antidepressant prescription rates, use of health services and outcomes among children, adolescents and young adults. AB - BACKGROUND: Regulatory bodies worldwide, including Health Canada, have issued warnings about prescribing antidepressants to children and adolescents. We sought to determine whether the Health Canada warning had the desired effects on prescribing patterns and outcomes and whether it had any unintended health consequences. METHODS: We examined data from prescription and health care databases representing more than 265 000 children, adolescents and young adults annually to determine changes in the rates of antidepressant prescription, use of health services and outcomes in these populations in the 9 years before and the 2 years after the Health Canada warning. We also examined the data for unintended changes in these rates among patients with anxiety disorders. We used young adults as the comparison group because they were not targeted by the warning. RESULTS: Following the warning, the rate of antidepressant prescriptions decreased among children and adolescents (relative risk [RR] 0.86, 95% confidence interval [CI] 0.81-0.91) and among young adults (RR 0.90, 95% CI 0.86-0.93). Ambulatory visits because of depression decreased among children and adolescents (RR 0.90, 95% CI 0.85-0.96) and young adults (RR 0.91, 95% CI 0.87-0.96). The rate of completed suicides among children and adolescents rose significantly after the warning (RR 1.25, 95% CI 1.08-1.44; annual rate per 1000 = 0.04 before and 0.15 after the warning). There was no equivalent change in the rate of completed suicides among young adults (RR 1.01, 95% CI 0.93-1.10; annual rate per 1000 = 0.15 before and 0.22 after the warning). Among patients with an anxiety disorder, the prescription rates did not change among children and adolescents, except for a decrease in the use of selective serotonin reuptake inhibitors other than fluoxetine, but the rates among young adults changed similar to the pattern of changes in the overall prescribing of antidepressants. There was also a significant decrease in the rate of physician visits because of anxiety disorders among young adults after the warning. INTERPRETATION: Health advisories and warnings issued by regulatory bodies may have unintended consequences on the provision of care, delivery of health services and clinical outcomes. Further efforts are required to ensure that health warnings do not result in unexpected harm. PMID- 18390945 TI - Battling depression. PMID- 18390947 TI - Asthma in Canada: missing the treatment targets. PMID- 18390946 TI - Health advisories: when good intentions go bad. PMID- 18390948 TI - The Canadian Academy of Health Sciences: coming of age at the right time. PMID- 18390949 TI - Obesity reduction and its possible consequences: what can we learn from Cuba's Special Period? PMID- 18390950 TI - Involving physicians in MedsCheck. PMID- 18390951 TI - Recruiting foreign-trained professionals. PMID- 18390952 TI - Recruiting foreign-trained professionals. PMID- 18390953 TI - Requiem for the sounds of silence. PMID- 18390954 TI - EuroSCORE predicts health-related quality of life after coronary artery bypass grafting. AB - Three hundred and two patients were evaluated for the EuroSCORE risk and health related quality of life (HRQoL) during three years after CABG as assessed by the 15D instrument. Both additive and logistic EuroSCORE correlated significantly with the 15D score at 6, 18 and 36 months. A clinically important increase > or =0.03 in the 15D score was achieved by 50.6% of patients at 6 months, 40.0% at 18 months and 35.9% at 36 months. The rates were similar among patients with increasing EuroSCORE at 6 and 18 months, but tended to decrease at 36 months in the highest EuroSCORE group (EuroSCORE 0-2: 46.8%; 3-5: 34.8%; and 6-14: 33.3%, respectively, P=0.13). Both additive (area under the receiver operating characteristic curve, AUC: 0.582, P=0.024) and logistic EuroSCORE (AUC: 0.575, P=0.039) were predictors of a significant increase of the 15D score. The best cut off value of the additive EuroSCORE for prediction of a clinically important improvement of the 15D score during 3-year follow-up was 3, as 46.7% of patients with EuroSCORE 0-3 and 30.1% of patients with a score >3 (P=0.006) improved clinically. The present study showed that the EuroSCORE also predicts long-term HRQoL after CABG. PMID- 18390955 TI - Intraoperative coronary angiography in postinfarction ventricular free wall rupture: how technology can change diagnostic and therapeutic timing. AB - Left ventricular free wall rupture often presents with an abrupt onset and rapidly progresses towards cardiogenic shock or electromechanical dissociation. The diagnostic pathway is still a matter of debate: echocardiography is commonly decisive but the assessment of coronary artery status is essential in order to optimize the surgical procedure. However, a preoperative coronary angiography could generate a dramatic delay of surgery. We report a case of a patient with a post-infarction left ventricular free wall rupture presenting with cardiac tamponade and cardiogenic shock who underwent emergency surgery. After cardiopulmonary bypass institution, an intraoperative coronary angiography was performed. Successful repair of the ventricular free wall rupture associated with a well-targeted surgical myocardial revascularization were carried out. This case illustrates how the development of technologically advanced hybrid operating rooms could lead to a new diagnostic and therapeutic approach to this potentially fatal complication. PMID- 18390956 TI - Graft repair of tracheo-innominate artery fistula following percutaneous tracheostomy. AB - Tracheo-innominate fistula (TIF) is a rare complication following percutaneous dilatational tracheostomy (PDT), occurring in < or =1% of cases. It usually develops three days to six weeks after the procedure and is fatal in the majority of cases, even after successful initial repair. We present a successfully treated case of TIF using a Goretex graft to replace the severely destroyed segment of the innominate artery. PMID- 18390957 TI - Percutaneously inserted central catheter for total parenteral nutrition in neonates: complications rates related to upper versus lower extremity insertion. AB - OBJECTIVE: The objective of this study was to compare the complication rates of upper versus lower extremity percutaneously inserted central catheters used for total parenteral nutrition in neonates. METHODS: During a 48-month study period, 396 neonates were identified as having had percutaneously inserted central venous catheters. A total of 370 catheters were inserted from the upper and 107 from the lower extremity. Data retrieved and analyzed were birth weight, gestational age, age at placement, duration in place, duration of total parenteral nutrition, type of infusates, catheter-related bloodstream infection, phlebitis, leakage, occlusion, necrotizing enterocolitis, intraventricular hemorrhage, serum creatinine, liver function tests, and length of hospitalization. RESULTS: The median birth weight and gestational age were 940 g and 28 weeks. The rate of catheter-related bloodstream infection was 11.6% for the upper and 9.3% in the lower extremity catheters. The most common organism was coagulase-negative Staphylococcus for both upper and lower extremity catheters and significantly higher with catheters from the upper extremity. Lower extremity catheters were in place longer, and the time from insertion to complication was also longer. The rate of cholestasis was higher for the upper extremity catheters. Multiple regression analysis showed that the most significant contributor to cholestasis was duration of time the catheters were in place and the duration of total parenteral nutrition administration. Receiver operating characteristics curve demonstrated higher sensitivity for duration of catheters in predicting cholestasis with duration of total parenteral nutrition being more specific. CONCLUSION: Lower extremity percutaneously inserted central venous catheters had lower rates of catheter-related bloodstream infection, longer time to first complication, and lower cholestasis despite longer duration of total parenteral nutrition. When possible, lower extremity inserted catheters should be used for the administration of total parenteral nutrition. PMID- 18390959 TI - Quality of general movements is related to white matter pathology in very preterm infants. AB - OBJECTIVE: Preterm infants are at an increased risk for abnormalities of general movements, which predict subsequent poor neurodevelopmental outcome. The cerebral lesion that predisposes the preterm infant to abnormal general movements remains unknown. The objective of this study was to determine the association between MRI defined cerebral abnormalities and general movements at 1 and 3 months' corrected age in infants who were born very preterm. METHODS: Eighty-six preterm infants (<30 weeks' gestation) were prospectively recruited and underwent brain MRI at term-equivalent age to investigate the relationship between qualitative white and gray matter pathology and abnormality of general movements. Standardized videotaped recordings of general movements were obtained at 1 and 3 months postterm (+/-1 week) and scored without knowledge of the MRI findings. At 1 month corrected age, general movements of a writhing character were classified as normal or abnormal (poor repertoire, cramped synchronized, or chaotic). At 3 months' corrected age, fidgety general movements were classified as present or absent. RESULTS: At 1 month, 53 (62%) infants had abnormal general movements, 46 of whom had poor repertoire general movements and 7 of whom had cramped synchronized general movements. At 3 months, 23 (25%) infants had absent fidgety movements. At both 1 and 3 months of age, consistently abnormal general movement classifications were related to increasing white matter abnormality on MRI. In contrast, there were no significant relationships between general movement classifications and gray matter abnormalities, either individually or in total. CONCLUSION: The significant relationships between general movements at 1 and 3 months and cerebral white matter abnormalities on MRI in the very preterm infant support the concept that abnormal general movements reflect white matter injury. PMID- 18390960 TI - Boosting the effectiveness of rehabilitative exercise training. PMID- 18390961 TI - To be or not to be: light at the end of the tunnel of career counseling for atopics. PMID- 18390958 TI - Neurodevelopmental outcome of extremely low birth weight infants with posthemorrhagic hydrocephalus requiring shunt insertion. AB - OBJECTIVE: We aimed to evaluate neurodevelopmental and growth outcomes among extremely low birth weight infants who had severe intraventricular hemorrhage that required shunt insertion compared with infants without shunt insertion. METHODS: Infants who were born in 1993-2002 with birth weights of 401 to 1000 g were enrolled in a very low birth weight registry at medical centers that participate in the National Institute of Child Health and Human Development Neonatal Research Network, and returned for follow-up at 18 to 22 months' corrected age were studied. Eighty-two percent of survivors completed follow-up, and 6161 children were classified into 5 groups: group 1, no intraventricular hemorrhage/no shunt (n = 5163); group 2, intraventricular hemorrhage grade 3/no shunt (n = 459); group 3, intraventricular hemorrhage grade 3/shunt (n = 103); group 4, intraventricular hemorrhage grade 4/no shunt (n = 311); and group 5, intraventricular hemorrhage grade 4/shunt (n = 125). Group comparisons were evaluated with chi(2) and Wilcoxon tests, and regression models were used to compare outcomes after adjustment for covariates. RESULTS: Children with severe intraventricular hemorrhage and shunts had significantly lower scores on the Bayley Scales of Infant Development IIR compared with children with no intraventricular hemorrhage and with children with intraventricular hemorrhage of the same grade and no shunt. Infants with shunts were at increased risk for cerebral palsy and head circumference at the <10th percentile at 18 months' adjusted age. Greatest differences were observed between children with shunts and those with no intraventricular hemorrhage on these outcomes. CONCLUSIONS: This large cohort study suggests that extremely low birth weight children with severe intraventricular hemorrhage that requires shunt insertion are at greatest risk for adverse neurodevelopmental and growth outcomes at 18 to 22 months compared with children with and without severe intraventricular hemorrhage and with no shunt. Long-term follow-up is needed to determine whether adverse outcomes persist or improve over time. PMID- 18390962 TI - Update in critical care 2007. PMID- 18390963 TI - Update in chronic obstructive pulmonary disease 2007. PMID- 18390964 TI - An official American Thoracic Society clinical policy statement: palliative care for patients with respiratory diseases and critical illnesses. PMID- 18390965 TI - Gender and prognosis of chronic obstructive pulmonary disease. PMID- 18390966 TI - Arterial stiffness and chronic obstructive pulmonary disease. PMID- 18390967 TI - American Society of Clinical Oncology Statement on minimum standards and exemplary attributes of clinical trial sites. AB - PURPOSE: To describe both minimum requirements for a site conducting quality clinical trials and attributes of an exemplary site. METHODS: Minimum requirements and exemplary attributes were selected based on literature review, prevailing regulatory requirements, and consensus among a group of community and academic clinical researchers. RESULTS: To provide guidance to oncologists who wish to conduct patient-oriented research, recommendations are made to assist in the development and implementation of high-quality research programs with the priority of protecting the welfare and rights of trial participants. A quality research site complies with the International Conference on Harmonisation (ICH) Good Clinical Practice (GCP) guidelines, the accepted international ethical and scientific quality standards for designing, conducting, recording, and reporting trials involving human participants. Recognizing that many research sites conduct clinical trials in compliance with accepted GCP standards, supplemental attributes of an exemplary research site that exceed the GCP criteria are also described. These attributes include diversification of clinical trial mix, high accrual activity, participation in the trial development process, maintenance of high educational standards, quality assurance, multidisciplinary involvement in the clinical trial process, and promotion of clinical trial awareness programs. CONCLUSION: Meeting the minimum criteria ensures conduct of quality clinical trials; however, some sites may wish to incorporate value-added attributes to exceed GCP compliance. These attributes are proposed as performance goals rather than requirements, recognizing that all sites conducting research will not necessarily meet each attribute but may still conduct high-quality clinical trials. PMID- 18390969 TI - Graft-versus-lymphoma effect for aggressive T-cell lymphomas in adults: a study by the Societe Francaise de Greffe de Moelle et de Therapie Cellulaire. AB - PURPOSE: Aggressive T-cell lymphomas (ATCLs) represent 10% to 15% of non Hodgkin's lymphomas (NHLs) in adults. ATCLs show a worse prognosis than B-cell lymphomas. PATIENTS AND METHODS: On behalf of the Societe Francaise de Greffe de Moelle et de Therapie Cellulaire, we conducted a retrospective analysis including 77 ATCL patients who underwent allogeneic stem-cell transplantation (alloSCT). RESULTS: The different diagnosis included anaplastic large-cell lymphoma (ALCL; n = 27), peripheral T-cell lymphoma not otherwise specified (PTCL-NOS; n = 27), angioimmunoblastic T-cell lymphoma (AITL; n = 11), hepatosplenic gamma/delta lymphoma (HSL; n = 3), T-cell granular lymphocytic leukemia (T-GLL; n = 1), nasal natural killer (NK)/T-cell lymphoma (nasal-NK/L; n = 3) or non-nasal NK/T-cell lymphoma (non-nasal-NK/L; n = 2), enteropathy-type T-cell (n = 1), and human T lymphotropic virus (HTLV)-1 lymphoma (n = 2). Fifty-seven patients received a myeloablative conditioning regimen. Donors were human leukocyte antigen (HLA) matched in 70 cases and related in 60 cases. Thirty-one patients were in complete remission (CR) at the time of alloSCT, whereas 26 were in partial response (PR). Five-year toxicity-related mortality (TRM) incidence was 33% (95% CI, 24% to 46%). The 5-year overall survival (OS) and event-free survival (EFS) rates were 57% (95% CI, 45% to 68%) and 53% (95% CI, 41% to 64%), respectively. In multivariate analysis, chemoresistant disease (stable, refractory, or progressing disease) at the time of alloSCT and the occurrence of severe grade 3 to 4 acute graft-versus-host disease (aGVHD) were the strongest adverse prognostic factors for OS (P = .03 and .03, respectively). Disease status at transplantation significantly influenced the 5-year EFS (P = .003), and an HLA-mismatched donor increased TRM (P = .04). CONCLUSION: We conclude that alloSCT is a potentially efficient therapy for NK/T lymphomas and is worth further investigation through prospective clinical trials. PMID- 18390968 TI - Phase I pharmacokinetic and pharmacodynamic study of the oral, small-molecule mitogen-activated protein kinase kinase 1/2 inhibitor AZD6244 (ARRY-142886) in patients with advanced cancers. AB - PURPOSE: To assess the tolerability, pharmacokinetics (PKs), and pharmacodynamics (PDs) of the mitogen-activated protein kinase kinase (MEK) 1/2 inhibitor AZD6244 (ARRY-142886) in patients with advanced cancer. PATIENTS AND METHODS: In part A, patients received escalating doses to determine the maximum-tolerated dose (MTD). In both parts, blood samples were collected to assess PK and PD parameters. In part B, patients were stratified by cancer type (melanoma v other) and randomly assigned to receive the MTD or 50% MTD. Biopsies were collected to determine inhibition of ERK phosphorylation, Ki-67 expression, and BRAF, KRAS, and NRAS mutations. RESULTS: Fifty-seven patients were enrolled. MTD in part A was 200 mg bid, but this dose was discontinued in part B because of toxicity. The 50% MTD (100 mg bid) was well tolerated. Rash was the most frequent and dose-limiting toxicity. Most other adverse events were grade 1 or 2. The PKs were less than dose proportional, with a median half-life of approximately 8 hours and inhibition of ERK phosphorylation in peripheral-blood mononuclear cells at all dose levels. Paired tumor biopsies demonstrated reduced ERK phosphorylation (geometric mean, 79%). Five of 20 patients demonstrated >or= 50% inhibition of Ki 67 expression, and RAF or RAS mutations were detected in 10 of 26 assessable tumor samples. Nine patients had stable disease (SD) for >or= 5 months, including two patients with SD for 19 (thyroid cancer) and 22 (uveal melanoma plus renal cancer) 28-day cycles. CONCLUSION: AZD6244 was well tolerated with target inhibition demonstrated at the recommended phase II dose. PK analyses supported twice-daily dosing. Prolonged SD was seen in a variety of advanced cancers. Phase II studies are ongoing. PMID- 18390970 TI - Influence of activation state of ErbB-2 (HER-2) on response to adjuvant cyclophosphamide, doxorubicin, and fluorouracil for stage II, node-positive breast cancer: study 8541 from the Cancer and Leukemia Group B. AB - PURPOSE: ErbB-2 (human epidermal growth factor receptor 2) overexpression may be predictive of relative resistance and/or sensitivity to specific chemotherapeutic agents. Results from a previous study from the Cancer and Leukemia Group B (CALGB 8541) demonstrated an interaction between ErbB-2 and increasing dose of adjuvant cyclophosphamide, doxorubicin, and fluorouracil (CAF) chemotherapy. Other studies have suggested that evaluation of the phosphorylated/activated form of ErbB-2 might be more precise in defining the impact of ErbB-2 in breast cancer. We have evaluated tumor tissue sections from CALGB 8541 patients to determine whether the interaction of ErbB-2 with CAF dose is dependent on ErbB-2 activation state, and whether phosphorylated ErbB-2 is an adverse prognostic factor in patients treated with CAF. PATIENTS AND METHODS: Patients were randomly assigned to one of three dosing regimens of CAF. Paraffin samples from 992 of 1,572 patients who participated in CALGB 8541 were available. Of the 570 tumors with any staining for ErbB-2, 488 had tissue available for assay for phosphorylated ErbB-2, which was performed by immunohistochemistry. RESULTS: Of 910 total assessable cases, 112 of 488 ErbB-2-positive cases (23%) stained positively for phosphorylated ErbB 2. The previously described interaction of dosing regimen of CAF with ErbB-2 was not dependent on phosphorylation status of ErbB-2. CONCLUSION: Monitoring phosphorylation of ErbB-2 with an antiphospho-ErbB-2 antibody did not add further precision to identifying those patients most likely to benefit from increased dose of anthracycline-based adjuvant chemotherapy. Favorable outcomes are observed in ErbB-2-overexpressing patients treated with high-dose CAF regardless of ErbB-2 phosphorylation state. PMID- 18390971 TI - EPIC: phase III trial of cetuximab plus irinotecan after fluoropyrimidine and oxaliplatin failure in patients with metastatic colorectal cancer. AB - PURPOSE: To determine whether adding cetuximab to irinotecan prolongs survival in patients with metastatic colorectal cancer (mCRC) previously treated with fluoropyrimidine and oxaliplatin. PATIENTS AND METHODS: This multicenter, open label, phase III study randomly assigned 1,298 patients with epidermal growth factor receptor-expressing mCRC who had experienced first-line fluoropyrimidine and oxaliplatin treatment failure to cetuximab (400 mg/m(2) day 1 followed by 250 mg/m(2) weekly) plus irinotecan (350 mg/m(2) every 3 weeks) or irinotecan alone. Primary end point was overall survival (OS); secondary end points included progression-free survival (PFS), response rate (RR), and quality of life (QOL). RESULTS: Median OS was comparable between treatments: 10.7 months (95% CI, 9.6 to 11.3) with cetuximab/irinotecan and 10.0 months (95% CI, 9.1 to 11.3) with irinotecan alone (hazard ratio [HR], 0.975; 95% CI, 0.854 to 1.114; P = .71). This lack of difference may have been due to post-trial therapy: 46.9% of patients assigned to irinotecan eventually received cetuximab (87.2% of those who did, received it with irinotecan). Cetuximab added to irinotecan significantly improved PFS (median, 4.0 v 2.6 months; HR, 0.692; 95% CI, 0.617 to 0.776; P 24 h) for cardiovascular causes; inappropriate shocks (two or more episodes); and sustained symptomatic AT lasting > 48 h. The outcome variable was a pre-specified score that corrected for clinical severity and follow-up duration. Three hundred and thirty-four patients were analysed (DC-ICD, n = 112; SC-ICD, n = 111; SC-simulated, n = 111). The mean left ventricular ejection fraction was 0.36 +/- 0.13, 69% were in functional class > or = II. CSAE occurred in 65 DC-ICD, 82 SC-ICD, and 84 SC-simulated patients. The outcome variable was 33% lower in the DC-ICD group (OR 0.31; 95% CI 0.14-0.67; P = 0.0028). Mortality was 4% in DC, 9% in SC, and 10% in SC simulated. CONCLUSION: In patients with a standard SC-ICD indication, DC-ICD was associated with less CSAE when compared with SC-ICD. PMID- 18390989 TI - Decreased insulin sensitivity in small for gestational age males treated with GH and preterm untreated males: a study in young adults. AB - BACKGROUND: Low birth weight and preterm birth are associated with growth delay as well as the development of insulin resistance. Insulin resistance is especially seen in subjects with catch-up growth. GH therapy induces growth in short subjects with low birth weight at term, but little is known about the long term effects on insulin sensitivity. GH therapy is now also proposed for preterms that remain short. METHODS: We investigated insulin sensitivity using the gold standard hyperinsulinemic-euglycemic clamp technique in 10 young adult males born small for gestational age (SGA) who had been treated with GH during childhood (GH) in comparison with 15 males born preterm AGA (premAGA), 13 males born preterm SGA (premSGA), and 15 males born at term with normal birth weight (CON). Furthermore, we investigated the presence of the metabolic syndrome. RESULTS: Insulin sensitivity was decreased in premAGA, premSGA, and GH subjects compared with CON males. The metabolic syndrome was not present in any of the groups. CONCLUSION: Insulin sensitivity is decreased in GH-treated SGA born males as well as in preterm born males. With respect to the SGA subjects, whether the difference results from perinatal-, postnatal-, or GH therapy-related factors are not known. With respect to the preterm born subjects, close surveillance is needed when commencing GH therapy. PMID- 18390990 TI - Effect of hypocaloric diet-induced weight loss in obese women on plasma apelin and adipose tissue expression of apelin and APJ. AB - OBJECTIVE: Apelin is a novel adipokine acting on APJ receptor, regulated by insulin and tumor necrosis factor-alpha (TNF-alpha) in adipose tissue (AT). Plasma apelin levels are increased in obese hyperinsulinemic subjects. The aim was to investigate whether the hypocaloric diet associated with weight loss modifies the elevated plasma apelin levels and the expression of apelin and APJ receptor in AT in obese women. DESIGN AND METHODS: Fasting plasma levels of apelin and TNF-alpha as well as mRNA levels of apelin and APJ in AT were measured before and after a 12-week hypocaloric weight-reducing diet in 20 obese women (body mass index (BMI) before diet 32.2+/-6.4 kg/m(2)). Twelve healthy women with a BMI of 20.7+/-0.6 kg/m(2) served as reference. RESULTS: Plasma levels of apelin and TNF-alpha were higher in obese compared with lean controls. The hypocaloric diet resulted in a significant decrease of BMI to 29.8+/-6.3 kg/m(2), plasma insulin (8.16+/-0.73 to 6.58+/-0.66 mU/l), apelin (369+/-25 pg/ml to 257+/-12 pg/ml), TNF-alpha levels (0.66+/-0.04 pg/ml to 0.56+/-0.04 pg/ml), and AT mRNAs of apelin and APJ. In addition, changes in AT mRNA apelin were related to changes in AT mRNA APJ levels. CONCLUSION: The hypocaloric diet associated with weight loss reduces the increased plasma and AT expression of apelin in obese women. This reduced apelin expression in AT could contribute to decreased circulating apelin levels. PMID- 18390991 TI - Centrally located body fat is related to appetitive hormones in healthy postmenopausal women. AB - OBJECTIVE: Body composition and energy homeostasis are thought to affect the appetitive hormones: adiponectin, leptin, insulin, and ghrelin. This study examined whether centrally located fat and/or overall adiposity were related to these appetitive hormones in healthy postmenopausal women. DESIGN: Overall and regional body composition was assessed by dual-energy X ray absorptiometry in relation to plasma adiponectin, serum leptin, serum insulin, and plasma ghrelin in 242 postmenopausal women. RESULTS: Regression analyses revealed that the androidal-to-gynoidal fat mass ratio (18.0%), age (3.2%), and white blood cell count (1.8%) accounted for 28% of the variability in adiponectin (F=22.2; P<0.0001); androidal (waist+hip) fat mass (66.0%), androidal fat mass(2) (6.2%), whole-body lean mass (2.2%), and age (0.8%) accounted for 69% of the variability in leptin (F=102.5; P<0.0001). Regression analyses revealed that sagittal abdominal diameter (8.4%), glucose (5.4%), white blood cell count (2.6%), and dietary omega-3 fatty acids (2.0%) accounted for 32% of the variability in insulin (F=20.8; P<0.0001); waist circumference (12.7%), hip lean mass (2.0%), and white blood cell count (1.9%) accounted for 26% of the variability in ghrelin (F=20.7; P<0.0001). Our results indicated that centralized fat mass was the primary contributor to these appetitive hormones in healthy postmenopausal women. CONCLUSION: Since central adiposity in postmenopausal women was related to appetitive hormones, minimizing weight gain during the menopausal transition may optimize appetitive hormones, thereby facilitating appetite control and weight maintenance. PMID- 18390992 TI - Mifepristone for ectopic ACTH secretion in metastatic endocrine carcinomas: report of two cases. AB - Ectopic adrenocorticotropin secretion (EAS) remains a therapeutic challenge whenever the tumor responsible for the syndrome is not amenable to curative resection. Two cases of EAS related to metastatic foregut-derived endocrine carcinomas led us to use mifepristone, an antagonist of both progesterone and glucocorticoids. Mifepristone clearly improved skin lesions and diabetes associated with hypercorticism. The beneficial effect lasted for about 10 months. In both cases, recurrent hypertension and hypokalemia eventually required adrenalectomy. PMID- 18390993 TI - Impact of total cumulative glucocorticoid dose on bone mineral density in patients with 21-hydroxylase deficiency. AB - OBJECTIVE: It remains controversial whether long-term glucocorticoids are charged of bone demineralization in patients with congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency. The aim of this study was to know whether cumulative glucocorticoid dose from the diagnosis in childhood to adulthood in patients with CAH had a negative impact on bone mineral density (BMD). DESIGN: This was a retrospective study. METHODS: Thirty-eight adult patients with classical and non-classical CAH were included. BMD was measured in the lumbar spine and femoral neck. Total cumulative glucocorticoid (TCG) and total average glucocorticoid (TAG) doses were calculated from pediatric and adult files. RESULTS: We showed a difference between final and target heights (-0.82+/-0.92 s.d. for women and -1.31+/-0.84 s.d. for men; P<0.001). Seventeen patients (44.7%) had bone demineralization (35.7% of women and 70% of men). The 28 women had higher BMD than the 10 men for lumbar (-0.26+/-1.20 vs -1.25+/-1.33 s.d.; P=0.02) and femoral T-scores (0.21+/-1.30 s.d. versus -1.08+/-1.10 s.d.; P=0.007). In the salt-wasting group, women were almost significantly endowed with a better BMD than men (P=0.053). We found negative effects of TCG, TAG on lumbar (P<0.001, P=0.002) and femoral T-scores (P=0.006, P<0.001), predominantly during puberty. BMI was protective on BMD (P=0.006). CONCLUSION: The TCG is an important factor especially during puberty for a bone demineralization in patients with 21 hydroxylase deficiency. The glucocorticoid treatment should be adapted particularly at this life period and preventive measures should be discussed in order to limit this effect. PMID- 18390994 TI - Predicting relapse in Crohn's disease: a biopsychosocial model. AB - BACKGROUND: Crohn's disease (CD) is a chronic relapsing inflammatory bowel disorder. Both biological and psychosocial factors may modulate the illness experience. AIM: The aim of this study was to identify clinical, biological and psychosocial parameters as predictors of clinical relapse in quiescent CD. METHODS: Patients in medically induced remission were followed prospectively for 1 year, or less if they relapsed. Disease characteristics were determined at baseline. Serum cytokines, anti-Saccharomyces cerevisiae antibodies, C-reactive protein (CRP), erythrocyte sedimentation rate and intestinal permeability were measured every 3 months. Psychological distress, perceived stress, minor life stressors and coping strategies were measured monthly. A time-dependent multivariate Cox regression model determined predictors of time to relapse. RESULTS: 101 patients (60 females, 41 males) were recruited. Fourteen withdrew and 37 relapsed. CRP (HR = 1.5 per 10 mg/l, 95% CI 1.1 to 1.9, p = 0.007), fistulising disease (HR = 3.2, 95% CI, 1.1 to 9.4, p = 0.04), colitis (HR = 3.5 95% CI 1.2 to 9.9, p = 0.02) and the interaction between perceived stress and avoidance coping (HR = 7.0 per 5 unit increase for both scales, 95% CI 2.3 to 21.8, p = 0.003) were predictors of earlier relapse. CONCLUSIONS: In quiescent CD, a higher CRP, fistulising disease behaviour and disease confined to the colon were independent predictors of relapse. Moreover, patients under conditions of low stress and who scored low on avoidance coping (ie, did not engage in social diversion or distraction) were least likely to relapse. This study supports a biopsychosocial model of CD exacerbation. PMID- 18390995 TI - Acquisition of the vacuolar ATPase proton pump and phagosome acidification are essential for escape of Francisella tularensis into the macrophage cytosol. AB - The Francisella tularensis-containing phagosome (FCP) matures to a late-endosome like phagosome prior to bacterial escape into the cytosols of macrophages, where bacterial proliferation occurs. Our data show that within the first 15 min after infection of primary human monocyte-derived macrophages (hMDMs), approximately 90% of the FCPs acquire the proton vacuolar ATPase (vATPase) pump and the lysomotropic dye LysoTracker, which concentrates in acidic compartments, similar to phagosomes harboring the Listeria monocytogenes control. The acquired proton vATPase pump and lysomotropic dye are gradually lost by 30 to 60 min postinfection, which coincides with bacterial escape into the cytosols of hMDMs. Colocalization of phagosomes harboring the iglD mutant with the vATPase pump and the LysoTracker dye was also transient, and the loss of colocalization was faster than that observed for the wild-type strain, which is consistent with the faster escape of the iglD mutant into the macrophage cytosol. In contrast, colocalization of both makers with phagosomes harboring the iglC mutant was persistent, which is consistent with fusion to the lysosomes and failure of the iglC mutant to escape into the macrophage cytosol. We have utilized a fluorescence microscopy-based phagosome integrity assay for differential labeling of vacuolar versus cytosolic bacteria, using antibacterial antibodies loaded into the cytosols of live hMDMs. We show that specific inhibition of the proton vATPase pump by bafilomycin A1 (BFA) blocks rapid bacterial escape into the cytosols of hMDMs, but 30% to 50% of the bacteria escape into the cytosol by 6 to 12 h after BFA treatment. The effect of BFA on the blocking of bacterial escape into the cytosol is completely reversible, as the bacteria escape after removal of BFA. We also show that the limited fusion of the FCP to lysosomes is not due to failure to recruit the late-endosomal fusion regulator Rab7. Therefore, within few minutes of its biogenesis, the FCP transiently acquires the proton vATPase pump to acidify the phagosome, and this transient acidification is essential for subsequent bacterial escape into the macrophage cytosol. PMID- 18390996 TI - Highly conserved surface proteins of oral spirochetes as adhesins and potent inducers of proinflammatory and osteoclastogenic factors. AB - Oral spirochetes include enormously heterogeneous Treponema species, and some have been implicated in the etiology of periodontitis. In this study, we characterized highly conserved surface proteins in four representative oral spirochetes (Treponema denticola, T. lecithinolyticum, T. maltophilum, and T. socranskii subsp. socranskii) that are homologs of T. pallidum Tp92, with opsonophagocytic potential and protective capacity against syphilis. Tp92 homologs of oral spirochetes had predicted signal peptides (20 to 31 amino acids) and molecular masses of 88 to 92 kDa for mature proteins. They showed amino acid sequence identities of 37.9 to 49.3% and similarities of 54.5 to 66.9% to Tp92. The sequence identities and similarities of Tp92 homologs of oral treponemes to one another were 41.6 to 71.6% and 59.9 to 85.6%, respectively. The tp92 gene homologs were successfully expressed in Escherichia coli, and the recombinant proteins were capable of binding to KB cells, an epithelial cell line, and inhibited the binding of the whole bacteria to the cells. Antiserum (the immunoglobulin G fraction) raised against a recombinant form of the T. denticola Tp92 homolog cross-reacted with homologs from three other species of treponemes. The Tp92 homologs stimulated various factors involved in inflammation and osteoclastogenesis, like interleukin-1beta (IL-1beta), tumor necrosis factor alpha, IL-6, prostaglandin E(2), and matrix metalloproteinase 9, in host cells like monocytes and fibroblasts. Our results demonstrate that Tp92 homologs of oral spirochetes are highly conserved and may play an important role in cell attachment, inflammation, and tissue destruction. The coexistence of various Treponema species in a single periodontal pocket and, therefore, the accumulation of multiple Tp92 homologs may amplify the pathological effect in periodontitis. PMID- 18390997 TI - Nasal immunization with Lactococcus lactis expressing the pneumococcal protective protein A induces protective immunity in mice. AB - Nisin-controlled gene expression was used to develop a recombinant strain of Lactococcus lactis that is able to express the pneumococcal protective protein A (PppA) on its surface. Immunodetection assays confirmed that after the induction with nisin, the PppA antigen was predictably and efficiently displayed on the cell surface of the recombinant strain, which was termed L. lactis PppA. The production of mucosal and systemically specific antibodies in adult and young mice was evaluated after mice were nasally immunized with L. lactis PppA. Immunoglobulin M (IgM), IgG, and IgA anti-PppA antibodies were detected in the serum and bronchoalveolar lavage fluid of adult and young mice, which showed that PppA expressed in L. lactis was able to induce a strong mucosal and systemic immune response. Challenge survival experiments demonstrated that immunization with L. lactis PppA was able to increase resistance to systemic and respiratory infection with different pneumococcal serotypes, and passive immunization assays of naive young mice demonstrated a direct correlation between anti-PppA antibodies and protection. The results presented in this study demonstrate three major characteristics of the effectiveness of nasal immunization with PppA expressed as a protein anchored to the cell wall of L. lactis: it elicited cross protective immunity against different pneumococcal serotypes, it afforded protection against both systemic and respiratory challenges, and it induced protective immunity in mice of different ages. PMID- 18390998 TI - Borrelia burgdorferi surface-localized proteins expressed during persistent murine infection are conserved among diverse Borrelia spp. AB - Borrelia burgdorferi, the causative agent of Lyme disease in the United States, regulates numerous genes encoding lipoproteins on linear plasmid 54 in response to environmental cues. We analyzed a subset of these genes/proteins that were historically categorized as paralogous gene family 54 (BBA64, BBA65, BBA66, BBA68, BBA69, BBA70, BBA71, and BBA73) and found that the expression of several genes was influenced by the sigma(N)-sigma(S) regulatory cascade at the level of transcription and protein synthesis. Moreover, we established in this and a previous study that BBA65, BBA66, BBA69, BBA71, and BBA73 are temporally expressed during persistent infection of immunocompetent mice, as determined by quantitative real time-PCR of ear tissue, by enzyme-linked immunosorbent assay, and by immunoblotting. Correspondingly, BBA65, BBA66, BBA71, and BBA73 proteins were detectable in infectious B. burgdorferi B31 isolates but undetectable in noninfectious isolates. BBA65, BBA66, BBA71, and BBA73 proteins were also found to partition into the Triton X-114 detergent phase and were sensitive to protease treatment of intact cells, indicating that they are membrane associated and surface localized. Lastly, Southern blotting and PCR with specific gene primer/probes for BBA64, BBA65, BBA66, BBA71, and BBA73 suggest that many of these genes are conserved among the B. burgdorferi sensu lato isolates and the relapsing-fever Borrelia species. Together, the data presented suggest that these genes may play a part in Borrelia infection and/or pathogenicity that could extend beyond the sensu lato group. PMID- 18390999 TI - Identification of Streptococcus sanguinis genes required for biofilm formation and examination of their role in endocarditis virulence. AB - Streptococcus sanguinis is one of the pioneers in the bacterial colonization of teeth and is one of the most abundant species in the oral biofilm called dental plaque. S. sanguinis is also the most common viridans group streptococcal species implicated in infective endocarditis. To investigate the association of biofilm and endocarditis, we established a biofilm assay and examined biofilm formation with a signature-tagged mutagenesis library of S. sanguinis. Four genes that have not previously been associated with biofilm formation in any other bacterium, purB, purL, thrB, and pyrE, were putatively identified as contributing to in vitro biofilm formation in S. sanguinis. By examining 800 mutants for attenuation in the rabbit endocarditis model and for reduction in biofilm formation in vitro, we found some mutants that were both biofilm defective and attenuated for endocarditis. However, we also identified mutants with only reduced biofilm formation or with only attenuation in the endocarditis model. This result indicates that the ability to form biofilms in vitro is not associated with endocarditis virulence in vivo in S. sanguinis. PMID- 18391000 TI - The innate immune response to Salmonella enterica serovar Typhimurium by macrophages is dependent on TREM2-DAP12. AB - Macrophage recognition of Salmonella enterica serovar Typhimurium leads to a cascade of signaling events, including the activation of Src family and Syk kinases and the production of reactive oxygen species (ROS), which are critical for host innate defense during early stages of bacterial infection. ROS production depends on the NADPH oxidase, but little is known about the innate immune receptors and proximal adapters that regulate Salmonella-induced ROS. Herein, we demonstrate that serovar Typhimurium induces ROS through a pathway that requires both triggering receptor expressed on myeloid cells 2 (TREM2) and DAP12. This pathway is highly analogous to the pathways utilized by Fc receptors and integrins to regulate ROS production. Oral infection of mice with serovar Typhimurium demonstrates that the DAP12-dependent pathway regulates cecal colonization during early stages of Salmonella infection. Thus, DAP12 is an important regulator of Salmonella-induced ROS production in macrophages, and TREM2 is essential for linking DAP12 to the innate response to serovar Typhimurium. PMID- 18391001 TI - Role of the mannose receptor in a murine model of Cryptococcus neoformans infection. AB - Cryptococcus neoformans is an encapsulated fungal pathogen with a predilection to infect persons with suppressed T-cell function. Cryptococcal mannoproteins (MP) are highly mannosylated antigens which elicit T-cell responses in infected mice and in convalescent patients. Key to the immunogenicity of MP is its capacity to bind to the conserved mannose receptor (MR), CD206, on dendritic cells (DCs). To test the role of the MR in the immune response to C. neoformans, wild-type and MR knockout (MR KO) mice were compared by using in vivo and ex vivo models of cryptococcosis. Following a pulmonary challenge with C. neoformans, MR KO mice died significantly faster than wild-type mice and had higher lung fungal burdens after 4 weeks of infection. Uptake of MP was similar when DCs obtained from wild type and MR KO mice were compared. Additionally, MP did not upregulate the maturation markers major histocompatibility complex class II, CD86, and CD40 in either wild-type or MR KO DCs. However, MP stimulated lymphoproliferation in CD4(+) T cells obtained from the peripheral lymph nodes of infected wild-type but not MR KO mice. These studies demonstrate a nonredundant role for the MR in the development of CD4(+) T-cell responses to MP and protection from C. neoformans. PMID- 18391002 TI - Inheritance of immune polarization patterns is linked to resistance versus susceptibility to Cryptococcus neoformans in a mouse model. AB - Genetic background variation between inbred strains accounts for different levels of susceptibility to Cryptococcus neoformans in the mouse infection model. To elucidate the inheritance of immunophenotypic traits and their associations with clearance outcomes during cryptococcal infection, we compared C57BL/6, BALB/c, and their first-generation hybrid, CB6F1 (F1), mice. Mice from each group were infected with C. neoformans (10(4) CFU) and analyzed at weekly intervals over a 6 week period. BALB/c mice progressively cleared the cryptococcal infection in the lungs and showed a Th1-skewed immune response: a Th1-shifted cytokine profile, modest lung pathology, and no significant elevation in the systemic immunoglobulin E (IgE) level. In contrast, C57BL/6 mice developed a chronic infection with a Th2-skewed immune response: a Th2-shifted cytokine profile, pulmonary eosinophilia, severe lung pathology, elevated serum IgE, fungemia, and cryptococcal dissemination in the central nervous system. F1 mice demonstrated intermediate resistance to C. neoformans, with a stronger resemblance to the immunophenotype of the resistant (BALB/c) mice. F1 mice also demonstrated enhanced pulmonary recruitment of lymphocytes, especially CD8(+) T cells, in comparison to both parental strains, suggesting positive heterosis. We conclude that the inheritance of traits responsible for early cytokine induction in the infected lungs and dendritic-cell maturation/activation status in draining nodes is responsible for the intermediate immune response polarization and clearance outcome observed initially in the lungs of F1 mice. The enhanced pulmonary lymphocyte recruitment could be responsible for a gradual shutdown of the undesirable Th2 arm of the immune response and subsequently improved anticryptococcal resistance in F1 mice. PMID- 18391003 TI - Respiratory Francisella tularensis live vaccine strain infection induces Th17 cells and prostaglandin E2, which inhibits generation of gamma interferon positive T cells. AB - Two key routes of Francisella tularensis infection are through the skin and airway. We wished to understand how the route of inoculation influenced the primary acute adaptive immune response. We show that an intranasal inoculation of the F. tularensis live vaccine strain (LVS) with a 1,000-fold-smaller dose than an intradermal dose results in similar growth kinetics and peak bacterial burdens. In spite of similar bacterial burdens, we demonstrate a difference in the quality, magnitude, and kinetics of the primary acute T-cell response depending on the route of inoculation. Further, we show that prostaglandin E(2) secretion in the lung is responsible for the difference in the gamma interferon (IFN-gamma) response. Intradermal inoculation led to a large number of IFN gamma(+) T cells 7 days after infection in both the spleen and the lung. In contrast, intranasal inoculation induced a lower number of IFN-gamma(+) T cells in the spleen and lung but an increased number of Th17 cells in the lung. Intranasal infection also led to a significant increase of prostaglandin E(2) (PGE(2)) in the bronchoalveolar lavage fluid. Inhibition of PGE(2) production with indomethacin treatment resulted in increased numbers of IFN-gamma(+) T cells and decreased bacteremia in the lungs of intranasally inoculated mice. This research illuminates critical differences in acute adaptive immune responses between inhalational and dermal infection with F. tularensis LVS mediated by the innate immune system and PGE(2). PMID- 18391004 TI - Extracellular signal-regulated kinase activation by Neisseria gonorrhoeae downregulates epithelial cell proapoptotic proteins Bad and Bim. AB - Neisseria gonorrhoeae expressing type IV pili (Tfp) activates extracellular signal-regulated kinase (ERK) and induces a cytoprotective state in the epithelial cell in a manner that is enhanced by pilT. As the ERK signaling pathway is well-known for its role in cytoprotection and cell survival, we tested the hypothesis that ERK is involved in producing this cytoprotective effect. Inhibiting ERK activation prior to infection attenuated the ability of these bacteria to induce cytoprotection. Activated ERK specifically targeted two proapoptotic Bcl-2 homology domain 3 (BH3)-only proteins, Bim and Bad, for downregulation at the protein level. Bim downregulation occurred through the proteasome. ERK, in addition, inactivated Bad by triggering its phosphorylation at Ser112. Finally, reducing the level of either Bad or Bim alone by small interfering RNA was sufficient to protect uninfected cells from staurosporine induced apoptosis. We conclude that Tfp-induced cytoprotection is due in part to ERK-dependent modification and/or downregulation of proapoptotic proteins Bad and Bim. PMID- 18391005 TI - Role of Porphyromonas gingivalis SerB in gingival epithelial cell cytoskeletal remodeling and cytokine production. AB - The SerB protein of Porphyromonas gingivalis is a HAD family serine phosphatase that plays a critical role in entry and survival of the organism in gingival epithelial cells. SerB is secreted by P. gingivalis upon contact with epithelial cells. Here it is shown by microarray analysis that SerB impacts the transcriptional profile of gingival epithelial cells, with pathways involving the actin cytoskeleton and cytokine production among those significantly overpopulated with differentially regulated genes. Consistent with the transcriptional profile, a SerB mutant of P. gingivalis exhibited defective remodeling of actin in epithelial cells. Interaction between gingival epithelial cells and isolated SerB protein resulted in actin rearrangement and an increase in the F/G actin ratio. SerB protein was also required for P. gingivalis to antagonize interleukin-8 accumulation following stimulation of epithelial cells with Fusobacterium nucleatum. SerB is thus capable of modulating host cell signal transduction that impacts the actin cytoskeleton and cytokine production. PMID- 18391006 TI - Protection against nasopharyngeal colonization by Streptococcus pneumoniae is mediated by antigen-specific CD4+ T cells. AB - CD4(+) T-cell-dependent acquired immunity confers antibody-independent protection against pneumococcal colonization. Since this mechanism is poorly understood for extracellular bacteria, we assessed the antigen specificity of the induction and recall of this immune response by using BALB/c DO11.10Rag(-/-) mice, which lack mature B and T cells except for CD4(+) T cells specific for the OVA(323-339) peptide derived from ovalbumin. Serotype 6B Streptococcus pneumoniae strain 603S and unencapsulated strain Rx1Delta lytA were modified to express OVA(323-339) as a fusion protein with surface protein A (PspA) (strains 603OVA(1) and Rx1Delta lytAOVA(1)) or with PspA, neuraminidase A, and pneumolysin (Rx1Delta lytAOVA(3)). Whole-cell vaccines (WCV) were made of ethanol-killed cells of Rx1Delta lytA plus cholera toxin (CT) adjuvant, of Rx1Delta lytAOVA(1) + CT (WCV-OVA(1)), and of Rx1Delta lytAOVA(3) + CT (WCV-OVA(3)). Mice intranasally immunized with WCV OVA(1), but not with WCV or CT alone, were protected against intranasal challenge with 603OVA(1). There was no protection against strain 603S in mice immunized with WCV-OVA(1). These results indicate antigen specificity of both immune induction and the recall response. Effector action was not restricted to antigen bearing bacteria since colonization by 603S was reduced in animals immunized with vaccines made of OVA-expressing strains when ovalbumin or killed Rx1Delta lytAOVA(3) antigen was administered around the time of challenge. CD4(+) T-cell mediated protection against pneumococcal colonization can be induced in an antigen-specific fashion and requires specific antigen for effective bacterial clearance, but this activity may extend beyond antigen-expressing bacteria. These results are consistent with the recruitment and/or activation of phagocytic or other nonspecific effectors by antigen-specific CD4(+) T cells. PMID- 18391007 TI - In LipL32, the major leptospiral lipoprotein, the C terminus is the primary immunogenic domain and mediates interaction with collagen IV and plasma fibronectin. AB - LipL32 is the major leptospiral outer membrane lipoprotein expressed during infection and is the immunodominant antigen recognized during the humoral immune response to leptospirosis in humans. In this study, we investigated novel aspects of LipL32. In order to define the immunodominant domains(s) of the molecule, subfragments corresponding to the N-terminal, intermediate, and C-terminal portions of the LipL32 gene were cloned and the proteins were expressed and purified by metal affinity chromatography. Our immunoblot results indicate that the C-terminal and intermediate domains of LipL32 are recognized by sera of patients with laboratory-confirmed leptospirosis. An immunoglobulin M response was detected exclusively against the LipL32 C-terminal fragment in both the acute and convalescent phases of illness. We also evaluated the capacity of LipL32 to interact with extracellular matrix (ECM) components. Dose-dependent, specific binding of LipL32 to collagen type IV and plasma fibronectin was observed, and the binding capacity could be attributed to the C-terminal portion of this molecule. Both heparin and gelatin could inhibit LipL32 binding to fibronectin in a concentration-dependent manner, indicating that the 30-kDa heparin-binding and 45-kDa gelatin-binding domains of fibronectin are involved in this interaction. Taken together, our results provide evidence that the LipL32 C terminus is recognized early in the course of infection and is the domain responsible for mediating interaction with ECM proteins. PMID- 18391008 TI - Streptococcus pneumoniae surface protein PcpA elicits protection against lung infection and fatal sepsis. AB - Previous studies have suggested that pneumococcal choline binding protein A (PcpA) is important for the full virulence of Streptococcus pneumoniae, and its amino acid sequence suggests that it may play a role in cellular adherence. PcpA is under the control of a manganese-dependent regulator and is only expressed at low manganese concentrations, similar to those found in the blood and lungs. PcpA expression is repressed under high manganese concentrations, similar to those found in secretions. In this study, we have demonstrated that PcpA elicits statistically significant protection in murine models of pneumonia and sepsis. In the model of pneumonia with each of four challenge strains, statistically fewer S. pneumoniae cells were recovered from the lungs of mice immunized with PcpA and alum versus mice immunized with alum only. The immunizations reduced the median CFU by 4- to 400-fold (average of 28-fold). In the model of sepsis using strain TIGR4, PcpA expression resulted in shorter times to become moribund and subcutaneous immunization with PcpA increased survival times of mice infected with wild-type PcpA-expressing pneumococci. PMID- 18391009 TI - Expression of the Mycobacterium tuberculosis acr-coregulated genes from the DevR (DosR) regulon is controlled by multiple levels of regulation. AB - Little is known about how Mycobacterium tuberculosis regulates gene expression in response to its host environment, despite its importance as a pathogen. We previously characterized 10 acr-coregulated genes (ACGs), all of which belong to the DevR (DosR) "dormancy" regulon, and identified one to three copies of a conserved 18-bp palindromic DNA motif in the promoter of each ACG family member. In the present study, we used base substitution analyses to assess the importance of individual motif copies and to identify additional regulatory sequences in five ACG promoters. Regulation of acr, acg, Rv2623, narK2, and Rv1738 was examined by using single-copy M. tuberculosis promoter-lacZ reporter constructs in Mycobacterium bovis BCG under conditions of ambient air versus hypoxia, each in shaking versus standing shallow culture conditions. We found that regulation of these ACG promoters is more heterogeneous than expected and is controlled at multiple levels. In addition to the positive regulation previously associated with DevR (DosR) and the 18-bp ACG motif, we identified negative regulation associated with sequences in the 5' untranslated regions of acg and Rv2623 and positive regulation associated with far upstream regulatory regions of narK2 and Rv1738. The importance of individual ACG motifs varied among the promoters examined, and Rv1738 was exceptional in that its ACG motif copies were associated with negative, rather than positive, regulation under some conditions. Further understanding of this important regulon requires the identification of additional regulators that compete and/or collaborate with DevR (DosR) to regulate its individual gene members. PMID- 18391010 TI - Immune recognition of Streptococcus pyogenes by dendritic cells. AB - Streptococcus pyogenes is one of the most frequent human pathogens. Recent studies have identified dendritic cells (DCs) as important contributors to host defense against S. pyogenes. The objective of this study was to identify the receptors involved in immune recognition of S. pyogenes by DCs. To determine whether Toll-like receptors (TLRs) were involved in DC sensing of S. pyogenes, we evaluated the response of bone marrow-derived DCs obtained from mice deficient in MyD88, an adapter molecule used by almost all TLRs, following S. pyogenes stimulation. Despite the fact that MyD88(-/-) DCs did not differ from wild-type DCs in the ability to internalize and kill S. pyogenes, the up-regulation of maturation markers, such as CD40, CD80, and CD86, and the production of inflammatory cytokines, such as interleukin-12 (IL-12), IL-6, and tumor necrosis factor alpha, were dramatically impaired in S. pyogenes-stimulated MyD88(-/-) DCs. These results suggest that signaling through TLRs is the principal pathway by which DCs sense S. pyogenes and become activated. Surprisingly, DCs deficient in signaling through each of the TLRs reported as potential receptors for gram positive cell components, such as TLR1, TLR2, TLR4, TLR9, and TLR2/6, were not impaired in the secretion of proinflammatory cytokines and the up-regulation of costimulatory molecules after S. pyogenes stimulation. In conclusion, our results exclude a major involvement of a single TLR or the heterodimer TLR2/6 in S. pyogenes sensing by DCs and argue for a multimodal recognition in which a combination of several different TLR-mediated signals is essential for a rapid and effective response to the pathogen. PMID- 18391011 TI - Characterization of fifty putative inclusion membrane proteins encoded in the Chlamydia trachomatis genome. AB - Although the Chlamydia trachomatis genome is predicted to encode 50 inclusion membrane proteins, only 18 have been experimentally localized in the inclusion membrane of C. trachomatis-infected cells. Using fusion proteins and anti-fusion protein antibodies, we have systematically evaluated all 50 putative inclusion membrane proteins for their localization in the infected cells, distribution patterns, and effects on subsequent chlamydial infection when expressed ectopically, as well as their immunogenicity during chlamydial infection in humans. Twenty-two of the 50 proteins were localized in the inclusion membrane, and 7 were detected inside the inclusions, while the location of the remaining 21 was not defined. Four (CT225, CT228, CT358, and CT440) of the 22 inclusion membrane-localized proteins were visualized in the inclusion membrane of Chlamydia-infected cells for the first time in the current study. The seven intra inclusion-localized proteins were confirmed to be chlamydial organism proteins in a Western blot assay. Further characterization of the 50 proteins revealed that neither colocalization with host cell endoplasmic reticulum nor inhibition of subsequent chlamydial infection by ectopically expressed proteins correlated with the inclusion membrane localization. Interestingly, antibodies from women with C. trachomatis urogenital infection preferentially recognized proteins localized in the inclusion membrane, and the immunodominant regions were further mapped to the region predicted to be on the cytoplasmic side of the inclusion membrane. These observations suggest that most of the inclusion membrane-localized proteins are both expressed and immunogenic during C. trachomatis infection in humans and that the cytoplasmic exposure may enhance the immunogenicity. PMID- 18391012 TI - Distinct expression and function of alternatively spliced Tbx5 isoforms in cell growth and differentiation. AB - Mutations in the T-box transcription factor Tbx5 cause Holt-Oram syndrome, an autosomal dominant disease characterized by a wide spectrum of cardiac and upper limb defects with variable expressivity. Tbx5 haploinsufficiency has been suggested to be the underlying mechanism, and experimental models are consistent with a dosage-sensitive requirement for Tbx5 in heart development. Here, we report that Tbx5 levels are regulated through alternative splicing that generates, in addition to the known 518-amino-acid protein, a C-terminal truncated isoform. This shorter isoform retains the capacity to bind DNA, but its interaction with Tbx5 collaborators such as GATA-4 is altered. In vivo, the two spliced isoforms are oppositely regulated in a temporal and growth factor dependent manner and are present in distinct DNA-binding complexes. The expression of the long isoform correlates with growth stimulation, and its reexpression in postnatal transgenic mouse hearts promotes hypertrophy. Conversely, the upregulation of the short but not the long isoform in C2C12 myoblasts leads to growth arrest and cell death. The results provide novel insight into posttranscriptional Tbx5 regulation and point to an important role not only in cell differentiation but also in cell proliferation and organ growth. The data may help analyze genotype-phenotype relations in patients with Holt-Oram syndrome. PMID- 18391013 TI - Transrepressive function of TLX requires the histone demethylase LSD1. AB - TLX is an orphan nuclear receptor (also called NR2E1) that regulates the expression of target genes by functioning as a constitutive transrepressor. The physiological significance of TLX in the cytodifferentiation of neural cells in the brain is known. However, the corepressors supporting the transrepressive function of TLX have yet to be identified. In this report, Y79 retinoblastoma cells were subjected to biochemical techniques to purify proteins that interact with TLX, and we identified LSD1 (also called KDM1), which appears to form a complex with CoREST and histone deacetylase 1. LSD1 interacted with TLX directly through its SWIRM and amine oxidase domains. LSD1 potentiated the transrepressive function of TLX through its histone demethylase activity as determined by a luciferase assay using a genomically integrated reporter gene. LSD1 and TLX were recruited to a TLX-binding site in the PTEN gene promoter, accompanied by the demethylation of H3K4me2 and deacetylation of H3. Knockdown of either TLX or LSD1 derepressed expression of the endogenous PTEN gene and inhibited cell proliferation of Y79 cells. Thus, the present study suggests that LSD1 is a prime corepressor for TLX. PMID- 18391014 TI - Targeted disruption of the basic Kruppel-like factor gene (Klf3) reveals a role in adipogenesis. AB - Kruppel-like factors (KLFs) recognize CACCC and GC-rich sequences in gene regulatory elements. Here, we describe the disruption of the murine basic Kruppel like factor gene (Bklf or Klf3). Klf3 knockout mice have less white adipose tissue, and their fat pads contain smaller and fewer cells. Adipocyte differentiation is altered in murine embryonic fibroblasts from Klf3 knockouts. Klf3 expression was studied in the 3T3-L1 cellular system. Adipocyte differentiation is accompanied by a decline in Klf3 expression, and forced overexpression of Klf3 blocks 3T3-L1 differentiation. Klf3 represses transcription by recruiting C-terminal binding protein (CtBP) corepressors. CtBPs bind NADH and may function as metabolic sensors. A Klf3 mutant that does not bind CtBP cannot block adipogenesis. Other KLFs, Klf2, Klf5, and Klf15, also regulate adipogenesis, and functional CACCC elements occur in key adipogenic genes, including in the C/ebpalpha promoter. We find that C/ebpalpha is derepressed in Klf3 and Ctbp knockout fibroblasts and adipocytes from Klf3 knockout mice. Chromatin immunoprecipitations confirm that Klf3 binds the C/ebpalpha promoter in vivo. These results implicate Klf3 and CtBP in controlling adipogenesis. PMID- 18391015 TI - MED1 phosphorylation promotes its association with mediator: implications for nuclear receptor signaling. AB - Mediator is a conserved multisubunit complex that acts as a functional interface between regulatory transcription factors and the general RNA polymerase II initiation apparatus. MED1 is a pivotal component of the complex that binds to nuclear receptors and a broad array of other gene-specific activators. Paradoxically, MED1 is found in only a fraction of the total cellular Mediator complexes, and the mechanisms regulating its binding to the core complex remain unclear. Here, we report that phosphorylation of MED1 by mitogen-activated protein kinase-extracellular signal-regulated kinase (MAPK-ERK) promotes its association with Mediator. We show that MED1 directly binds to the MED7 subunit and that ERK phosphorylation of MED1 enhances this interaction. Interestingly, we found that both thyroid and steroid hormones stimulate MED1 phosphorylation in vivo and that MED1 phosphorylation is required for its nuclear hormone receptor coactivator activity. Finally, we show that MED1 phosphorylation by ERK enhances thyroid hormone receptor-dependent transcription in vitro. Our findings suggest that ERK phosphorylation of MED1 is a regulatory mechanism that promotes MED1 association with Mediator and, as such, may facilitate a novel feed-forward action of nuclear hormones. PMID- 18391016 TI - The neogenin intracellular domain regulates gene transcription via nuclear translocation. AB - Neogenin is a multifunctional receptor implicated in axon navigation, neuronal differentiation, morphogenesis, and cell death. Very little is known about signaling downstream of neogenin. Because we found that the neogenin intracellular domain (NeICD) interacts with nuclear proteins implicated in transcription regulation, we investigated further whether neogenin signals similarly to the Notch receptor. We show here that neogenin is cleaved by gamma secretase, an event that releases the complete NeICD. We also describe that NeICD is located at the nucleus, a feature regulated through a balance between nuclear import and export. NeICD triggers gene reporter transactivation and associates with nuclear chromatin. Direct transcriptional targets of NeICD were determined and were shown to be up-regulated in the presence of neogenin ligand. Together, we reveal here a novel aspect of neogenin signaling that relies on the direct implication of its intracellular domain in transcriptional regulation. PMID- 18391017 TI - JunB breakdown in mid-/late G2 is required for down-regulation of cyclin A2 levels and proper mitosis. AB - JunB, a member of the AP-1 family of dimeric transcription factors, is best known as a cell proliferation inhibitor, a senescence inducer, and a tumor suppressor, although it also has been attributed a cell division-promoting activity. Its effects on the cell cycle have been studied mostly in G1 and S phases, whereas its role in G2 and M phases still is elusive. Using cell synchronization experiments, we show that JunB levels, which are high in S phase, drop during mid to late G2 phase due to accelerated phosphorylation-dependent degradation by the proteasome. The forced expression of an ectopic JunB protein in late G2 phase indicates that JunB decay is necessary for the subsequent reduction of cyclin A2 levels in prometaphase, the latter event being essential for proper mitosis. Consistently, abnormal JunB expression in late G2 phase entails a variety of mitotic defects. As these aberrations may cause genetic instability, our findings contrast with the acknowledged tumor suppressor activity of JunB and reveal a mechanism by which the deregulation of JunB might contribute to tumorigenesis. PMID- 18391018 TI - Cyclic AMP (cAMP)-mediated stimulation of adipocyte differentiation requires the synergistic action of Epac- and cAMP-dependent protein kinase-dependent processes. AB - Cyclic AMP (cAMP)-dependent processes are pivotal during the early stages of adipocyte differentiation. We show that exchange protein directly activated by cAMP (Epac), which functions as a guanine nucleotide exchange factor for the Ras like GTPases Rap1 and Rap2, was required for cAMP-dependent stimulation of adipocyte differentiation. Epac, working via Rap, acted synergistically with cAMP dependent protein kinase (protein kinase A [PKA]) to promote adipogenesis. The major role of PKA was to down-regulate Rho and Rho-kinase activity, rather than to enhance CREB phosphorylation. Suppression of Rho-kinase impaired proadipogenic insulin/insulin-like growth factor 1 signaling, which was restored by activation of Epac. This interplay between PKA and Epac-mediated processes not only provides novel insight into the initiation and tuning of adipocyte differentiation, but also demonstrates a new mechanism of cAMP signaling whereby cAMP uses both PKA and Epac to achieve an appropriate cellular response. PMID- 18391019 TI - Autoregulation of Npl3, a yeast SR protein, requires a novel downstream region and serine phosphorylation. AB - Npl3 is an SR-like protein with documented roles in mRNA export and transcription termination. Maintaining appropriate levels of Npl3 protein is critical for cell survival. Here we show that Npl3 negatively regulates its own expression via modulation of its mRNA levels. By creating gene chimeras, we demonstrate that the region downstream of the coding sequence of Npl3 is necessary and sufficient to confer regulation. The use of different polyadenylation sites in this region results in at least two stable RNAs; read-through of these sites causes the formation of 3'-extended RNAs that are highly unstable and therefore largely unproductive. Increasing the amount of Npl3 protein promotes read-through. Notably, the loss of Npl3 phosphorylation promotes the use of the productive polyadenylation sites, resulting in elevated levels of Npl3 protein. We propose that proper levels of Npl3 protein are achieved by a negative feedback loop in which phosphorylated Npl3 suppresses efficient recognition of the productive processing signals in its own transcript. PMID- 18391020 TI - Sir2 silences gene transcription by targeting the transition between RNA polymerase II initiation and elongation. AB - It is well accepted that for transcriptional silencing in budding yeast, the evolutionarily conserved lysine deacetylase Sir2, in concert with its partner proteins Sir3 and Sir4, establishes a chromatin structure that prevents RNA polymerase II (Pol II) transcription. However, the mechanism of repression remains controversial. Here, we show that the recruitment of Pol II, as well as that of the general initiation factors TBP and TFIIH, occurs unimpeded to the silent HMRa1 and HMLalpha1/HMLalpha2 mating promoters. This, together with the fact that Pol II is Ser5 phosphorylated, implies that SIR-mediated silencing is permissive to both preinitiation complex (PIC) assembly and transcription initiation. In contrast, the occupancy of factors critical to both mRNA capping and Pol II elongation, including Cet1, Abd1, Spt5, Paf1C, and TFIIS, is virtually abolished. In agreement with this, efficiency of silencing correlates not with a restriction in Pol II promoter occupancy but with a restriction in capping enzyme recruitment. These observations pinpoint the transition between polymerase initiation and elongation as the step targeted by Sir2 and indicate that transcriptional silencing is achieved through the differential accessibility of initiation and capping/elongation factors to chromatin. We compare Sir2-mediated transcriptional silencing to a second repression mechanism, mediated by Tup1. In contrast to Sir2, Tup1 prevents TBP, Pol II, and TFIIH recruitment to the HMLalpha1 promoter, thereby abrogating PIC formation. PMID- 18391021 TI - Binding of DAZAP1 and hnRNPA1/A2 to an exonic splicing silencer in a natural BRCA1 exon 18 mutant. AB - A disease-causing G-to-T transversion at position +6 of BRCA1 exon 18 induces exclusion of the exon from the mRNA and, as has been suggested by in silico analysis, disrupts an ASF/SF2-dependent splicing enhancer. We show here using a pulldown assay with an internal standard that wild-type (WT) and mutant T6 sequences displayed similar ASF/SF2 binding efficiencies, which were significantly lower than that of a typical exonic splicing enhancer derived from the extra domain A exon of fibronectin. Overexpression or small interfering RNA (siRNA)-mediated depletion of ASF/SF2 did not affect the splicing of a WT BRCA1 minigene but resulted in an increase and decrease of T6 exon 18 inclusion, respectively. Furthermore, extensive mutation analysis using hybrid minigenes indicated that the T6 mutant creates a sequence with a prevalently inhibitory function. Indeed, RNA-protein interaction and siRNA experiments showed that the skipping of T6 BRCA1 exon 18 is due to the creation of a splicing factor dependent silencer. This sequence specifically binds to the known repressor protein hnRNPA1/A2 and to DAZAP1, the involvement of which in splicing inhibition we have demonstrated. Our results indicate that the binding of the splicing factors hnRNPA1/A2 and DAZAP1 is the primary determinant of T6 BRCA1 exon 18 exclusion. PMID- 18391022 TI - A novel protein, Luman/CREB3 recruitment factor, inhibits Luman activation of the unfolded protein response. AB - Luman/CREB3 (also called LZIP) is an endoplasmic reticulum (ER)-bound cellular transcription factor. It has been implicated in the mammalian unfolded protein response (UPR), as well as herpes simplex virus reactivation from latency in sensory neurons. Here, we report the identification of a novel Luman recruitment factor (LRF). Like Luman, LRF is a UPR-responsive basic-region leucine zipper protein that is prone to proteasomal degradation. Being a highly unstable protein, LRF interacts with Luman through the leucine zipper region and promotes Luman degradation. LRF was found to recruit the nuclear form of Luman to discrete nuclear foci, which overlap with the nuclear receptor coactivator GRIP1 bodies, and repress the transactivation activity of Luman. Compared to LRF+/+ mouse embryonic fibroblast (MEF) cells, the levels of CHOP, EDEM, and Herp were elevated in LRF-/- MEF cells. We propose that LRF is a negative regulator of the UPR. For Luman, it may represent another level of regulation following Luman proteolytic cleavage on the ER and nuclear translocation. In addition to inducing rapid Luman turnover, LRF may repress the transactivation potential of Luman by sequestering it in the LRF nuclear bodies away from key cofactors (such as HCF-1) that are required for transcriptional activation. PMID- 18391023 TI - PTEN represses RNA polymerase III-dependent transcription by targeting the TFIIIB complex. AB - PTEN, a tumor suppressor whose function is frequently lost in human cancers, possesses a lipid phosphatase activity that represses phosphatidylinositol 3 kinase (PI3K) signaling, controlling cell growth, proliferation, and survival. The potential for PTEN to regulate the synthesis of RNA polymerase (Pol) III transcription products, including tRNAs and 5S rRNAs, was evaluated. The expression of PTEN in PTEN-deficient cells repressed RNA Pol III transcription, whereas decreased PTEN expression enhanced transcription. Transcription repression by PTEN was uncoupled from PTEN-mediated effects on the cell cycle and was independent of p53. PTEN acts through its lipid phosphatase activity, inhibiting the PI3K/Akt/mTOR/S6K pathway to decrease transcription. PTEN, through the inactivation of mTOR, targets the TFIIIB complex, disrupting the association between TATA-binding protein and Brf1. Kinetic analysis revealed that PTEN initially induces a decrease in the serine phosphorylation of Brf1, leading to a selective reduction in the occupancy of all TFIIIB subunits on tRNA(Leu) genes, whereas prolonged PTEN expression results in the enhanced serine phosphorylation of Bdp1. Together, these results demonstrate a new class of genes regulated by PTEN through its ability to repress the activation of PI3K/Akt/mTOR/S6K signaling. PMID- 18391024 TI - Synthetic lethal screens identify gene silencing processes in yeast and implicate the acetylated amino terminus of Sir3 in recognition of the nucleosome core. AB - Dot1 methylates histone H3 lysine 79 (H3K79) on the nucleosome core and is involved in Sir protein-mediated silencing. Previous studies suggested that H3K79 methylation within euchromatin prevents nonspecific binding of the Sir proteins, which in turn facilitates binding of the Sir proteins in unmethylated silent chromatin. However, the mechanism by which the Sir protein binding is influenced by this modification is unclear. We performed genome-wide synthetic genetic array (SGA) analysis and identified interactions of DOT1 with SIR1 and POL32. The synthetic growth defects found by SGA analysis were attributed to the loss of mating type identity caused by a synthetic silencing defect. By using epistasis analysis, DOT1, SIR1, and POL32 could be placed in different pathways of silencing. Dot1 shared its silencing phenotypes with the NatA N-terminal acetyltransferase complex and the conserved N-terminal bromo adjacent homology (BAH) domain of Sir3 (a substrate of NatA). We classified all of these as affecting a common silencing process, and we show that mutations in this process lead to nonspecific binding of Sir3 to chromatin. Our results suggest that the BAH domain of Sir3 binds to histone H3K79 and that acetylation of the BAH domain is required for the binding specificity of Sir3 for nucleosomes unmethylated at H3K79. PMID- 18391025 TI - Negational categorization and intergroup behavior. AB - Individuals define themselves, at times, as who they are (e.g., a psychologist) and, at other times, as who they are not (e.g., not an economist). Drawing on social identity, optimal distinctiveness, and balance theories, four studies examined the nature of negational identity relative to affirmational identity. One study explored the conditions that increase negational identification and found that activating the need for distinctiveness increased the accessibility of negational identities. Three additional studies revealed that negational categorization increased outgroup derogation relative to affirmational categorization and the authors argue that this effect is at least partially due to a focus on contrasting the self from the outgroup under negational categorization. Consistent with this argument, outgroup derogation following negational categorization was mitigated when connections to similar others were highlighted. By distinguishing negational identity from affirmational identity, a more complete picture of collective identity and intergroup behavior can start to emerge. PMID- 18391026 TI - Population pharmacokinetics of rifampin in pulmonary tuberculosis patients, including a semimechanistic model to describe variable absorption. AB - This article describes the population pharmacokinetics of rifampin in South African pulmonary tuberculosis patients. Three datasets containing 2,913 rifampin plasma concentration-time data points, collected from 261 South African pulmonary tuberculosis patients aged 18 to 72 years and weighing 28.5 to 85.5 kg and receiving regular daily treatment that included administration of rifampin (450 to 600 mg) for at least 10 days, were pooled. A compartmental pharmacokinetic model was developed using nonlinear mixed-effects modeling. Variability in the shape of the absorption curve was described using a flexible transit compartment model, in which a delay in the onset of absorption and a gradually changing absorption rate were modeled as the passage of drug through a chain of hypothetical compartments, ultimately reaching the absorption compartment. A previously described implementation was extended to allow its application to multiple-dosing data. The typical population estimate of oral clearance was 19.2 liters x h(-1), while the volume of distribution was estimated to be 53.2 liters. Interindividual variability was estimated to be 52.8% for clearance and 43.4% for volume of distribution. Interoccasional variability was estimated for CL/F (22.5%) and mean transit time during absorption (67.9%). The use of single-drug formulations was found to increase both the mean transit time (by 104%) and clearance (by 23.6%) relative to fixed-dose-combination use. A strong correlation between clearance and volume of distribution suggested substantial variability in bioavailability, which could have clinical implications, given the dependence of treatment effectiveness on exposure. The final model successfully described rifampin pharmacokinetics in the population studied and is suitable for simulation in this context. PMID- 18391027 TI - In vitro antifungal susceptibility and molecular characterization of clinical isolates of Fusarium verticillioides (F. moniliforme) and Fusarium thapsinum. AB - A microdilution method was used to test 11 antifungal drugs against clinical isolates of Fusarium thapsinum and three different phylogenetic clades of Fusarium verticillioides that were characterized by sequencing a region of the beta-tubulin gene. Terbinafine was the most-active drug against both species, followed by posaconazole against F. verticillioides. PMID- 18391028 TI - Increasing incidence of linezolid-intermediate or -resistant, vancomycin resistant Enterococcus faecium strains parallels increasing linezolid consumption. AB - Clinical enterococcal resistance to linezolid is defined by the presence of the G2576T mutation. We evaluated the incidence of genetically proven linezolid resistance among vancomycin-resistant Enterococcus faecium strains and linezolid consumption for a possible association. A relationship was found (r(2) = 0.73, P = 0.03) and predicts increasing resistance with current trends of linezolid use. PMID- 18391030 TI - In vitro activities of linezolid against clinical isolates of Mycobacterium tuberculosis complex isolated in Taiwan over 10 years. AB - Significant increases in the MIC(90)s of linezolid in multidrug-resistant Mycobacterium tuberculosis isolates were seen between the baseline period of 2001 to 2003 (0.5 microg/ml) and 2004 (2 microg/ml). The MICs were 4 microg/ml in three strains. Both fluoroquinolones (except levofloxacin) and kanamycin were found to have statistically significant degrees of concordance with linezolid. PMID- 18391029 TI - Inhibition of herpes simplex virus type 1 infection by cationic beta-peptides. AB - Previously, it was shown that cationic alpha-peptides derived from the human immunodeficiency virus TAT protein transduction domain blocked herpes simplex virus type 1 (HSV-1) entry. We now show that cationic oligomers of beta-amino acids ("beta-peptides") inhibit HSV-1 infection. Among three cationic beta peptides tested, the most effective inhibition was observed for the one with a strong propensity to adopt a helical conformation in which cationic and hydrophobic residues are segregated from one another ("globally amphiphilic helix"). The antiviral effect was not cell type specific. Inhibition of virus infection by the beta-peptides occurred at the postattachment penetration step, with a 50% effective concentration of 3 muM for the most-effective beta-peptide. The beta-peptides did not inactivate virions in solution, nor did they induce resistance to infection when cells were pretreated with the beta-peptides. The beta-peptides showed little if any toxicity toward Vero cells. These results raise the possibility that cationic beta-peptides may be useful antiviral agents for HSV-1 and demonstrate the potential of beta-peptides as novel antiviral drugs. PMID- 18391031 TI - In vitro activity of anidulafungin against Candida albicans biofilms. AB - We tested the activity of anidulafungin against 30 Candida albicans isolates. The planktonic MICs for 50 and 90% of the isolates tested (MIC(50) and MIC(90)) were < or =0.03 and 0.125 microg/ml, respectively (MIC range, < or =0.03 to 2 microg/ml). The sessile MIC(50) and MIC(90) were < or =0.03 and < or =0.03 microg/ml, respectively (MIC range, < or =0.03 to >16 microg/ml). PMID- 18391032 TI - First report of cfr-mediated resistance to linezolid in human staphylococcal clinical isolates recovered in the United States. AB - Linezolid resistance has dominantly been mediated by mutations in 23S rRNA or ribosomal protein L4 genes. Recently, cfr has demonstrated the ability to produce a phenotype of resistance to not only oxazolidinones, but also other antimicrobial classes (phenicols, lincosamides, pleuromutilins, and streptogramin A). We describe the first detection of cfr-mediated linezolid resistance in Staphylococcus aureus and Staphylococcus epidermidis recovered from human infection cases monitored during the 2007 LEADER Program. PMID- 18391033 TI - Potency of SMP-601, a novel carbapenem, in hematogenous murine bronchopneumonia caused by methicillin-resistant and vancomycin-intermediate Staphylococcus aureus. AB - We compared the potency of SMP-601, a novel carbapenem, with that of vancomycin in a murine model of hematogenous bronchopneumonia infection caused by methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-intermediate S. aureus (VISA). The MICs of SMP-601 and vancomycin against MRSA were 2 and 1 mug/ml, respectively, while those against VISA were 2 and 8 mug/ml, respectively. Treatment with SMP-601 resulted in a significant decrease in the number of viable bacteria in the MRSA infection model (control, 100 mg/kg vancomycin, and 100 mg/kg SMP-601, 8.42 +/- 0.50, 5.29 +/- 0.71, and 5.50 +/- 0.58 log CFU/lung, respectively,) and in the VISA infection model (control, 100 mg/kg vancomycin, and 100 mg/kg SMP-601, 9.64 +/- 0.63, 8.72 +/- 0.45, 7.42 +/- 0.14 log CFU/lung) (mean +/- standard error of the mean). The survival rate in the VISA infection model treated with SMP-601 (70%) was significantly higher than those in the other two groups (20% for vancomycin and 0% for control; P < 0.05). Histopathological examination revealed that inflammatory changes in the SMP-601-treated group were less marked than in the other two groups. The results of pharmacokinetic pharmacodynamic analysis supported the results of the bacteriological, histopathological and survival studies. Our results demonstrate the potency of SMP-601 against MRSA and VISA in murine hematogenous pulmonary infection. PMID- 18391034 TI - Multivalent presentation of antihantavirus peptides on nanoparticles enhances infection blockade. AB - Viral entry into susceptible host cells typically results from multivalent interactions between viral surface proteins and host entry receptors. In the case of Sin Nombre virus (SNV), a New World hantavirus that causes hantavirus cardiopulmonary syndrome, infection involves the interaction between viral membrane surface glycoproteins and the human integrin alpha(v)beta(3). Currently, there are no therapeutic agents available which specifically target SNV. To address this problem, we used phage display selection of cyclic nonapeptides to identify peptides that bound SNV and specifically prevented SNV infection in vitro. We synthesized cyclic nonapeptides based on peptide sequences of phage demonstrating the strongest inhibition of infection, and in all cases, the isolated peptides were less effective at blocking infection (9.0% to 27.6% inhibition) than were the same peptides presented by phage (74.0% to 82.6% inhibition). Since peptides presented by the phage were pentavalent, we determined whether the identified peptides would show greater inhibition if presented in a multivalent format. We used carboxyl linkages to conjugate selected cyclic peptides to multivalent nanoparticles and tested infection inhibition. Two of the peptides, CLVRNLAWC and CQATTARNC, showed inhibition that was improved over that of the free format when presented on nanoparticles at a 4:1 nanoparticle-to-virus ratio (9.0% to 32.5% and 27.6% to 37.6%, respectively), with CQATTARNC inhibition surpassing 50% when nanoparticles were used at a 20:1 ratio versus virus. These data illustrate that multivalent inhibitors may disrupt polyvalent protein-protein interactions, such as those utilized for viral infection of host cells, and may represent a useful therapeutic approach. PMID- 18391035 TI - Mechanism of inhibition of human immunodeficiency virus type 1 reverse transcriptase by a stavudine analogue, 4'-ethynyl stavudine triphosphate. AB - 2',3'-Didehydro-3'-deoxy-4'-ethynylthymidine (4'-Ed4T), a recently discovered nucleoside reverse transcriptase (RT) inhibitor, exhibits 5- to 10-fold-higher activity against human immunodeficiency virus type 1 (HIV-1) and less cytotoxicity than does its parental compound d4T (stavudine). Using steady-state kinetic approaches, we have previously shown that (i) 4'-ethynyl-d4T triphosphate (4'-Ed4TTP) inhibits HIV-1 RT more efficiently than d4TTP does and (ii) its inhibition efficiency toward the RT M184V mutant is threefold less than that toward wild-type (wt) RT. In this study we used pre-steady-state kinetic approaches in an attempt to understand its mechanism of inhibition. With wt and the M184V mutant RTs, 4'-Ed4TTP has three- to fivefold-lower K(d) (dissociation constant) values than d4TTP, while d4TTP has up to eightfold-higher K(d) values than dTTP. Inhibition is more effective in DNA replication with RNA template than with DNA template. In general, the M184V mutant exhibits poorer binding for all three nucleoside triphosphates than does wt RT. The structural basis for the lower binding affinity of d4TTP than of dTTP could be the lack of hydrogen bonds from the missing 3'-hydroxyl group in d4TTP to the backbone amide of Y115 and also to the side chain of Q151. The structural basis for the higher binding affinity of 4'-Ed4TTP than of d4TTP could be the additional binding of the 4' ethynyl group in a preformed hydrophobic pocket by A114, Y115, M184, F160, and part of D185. PMID- 18391037 TI - Posaconazole against Candida glabrata isolates with various susceptibilities to fluconazole. AB - We investigated the in vitro activities of posaconazole (POS), fluconazole (FLC), amphotericin B (AMB), and caspofungin (CAS) against four clinical isolates of Candida glabrata with various susceptibilities to FLC (FLC MICs ranging from 1.0 to >64 microg/ml). POS MICs ranged from < or =0.03 to 0.5 microg/ml; AMB MICs ranged from 0.25 to 2.0 microg/ml, while CAS MICs ranged from 0.03 to 0.25 microg/ml. When FLC MICs increased, so did POS MICs, although we did not observe any isolate with a POS MIC greater than 0.5 mug/ml. Time-kill experiments showed that POS, FLC, and CAS were fungistatic against all isolates, while AMB at eight times the MIC was fungicidal against three out of four isolates of C. glabrata tested. Then, we investigated the activity of POS in an experimental model of disseminated candidiasis using three different isolates of C. glabrata: one susceptible to FLC (S; FLC MICs ranging from 1.0 to 4.0 microg/ml; POS MIC of < or =0.03 microg/ml), one susceptible in a dose-dependent manner (SDD; FLC MICs ranging from 32 to 64 microg/ml; POS MICs ranging from 0.125 to 0.25 microg/ml), and another one resistant to FLC (R; FLC MIC of >64 microg/ml; POS MIC of 0.5 microg/ml). FLC significantly reduced the kidney burden of mice infected with the S strain (P = 0.0070) but not of those infected with the S-DD and R strains. POS was significantly effective against all three isolates at reducing the kidney fungal burden with respect to the controls (P ranging from 0.0003 to 0.029). In conclusion, our data suggest that POS may be a useful option in the management of systemic infections caused by C. glabrata. Additionally, the new triazole may be a therapeutic option in those cases where an FLC-resistant isolate is found to retain a relatively low POS MIC. PMID- 18391036 TI - Production of enterocins L50A, L50B, and IT, a new enterocin, by Enterococcus faecium IT62, a strain isolated from Italian ryegrass in Japan. AB - Enterococcus faecium IT62, isolated from ryegrass in Japan, was shown to produce three different bacteriocins, two of which had molecular masses and amino acid sequences that corresponded to those of enterocin L50A and enterocin L50B. These peptides existed, however, as chemically modified forms that were either N formylated or N formylated and oxidized at Met(24). The third bacteriocin, named enterocin IT, had a molecular mass of 6,390 Da, was made up of 54 amino acids, and did not correspond to any known bacteriocin. However, enterocin IT was identical to the C-terminal part of the 16-amino-acid-longer bacteriocin 32 (T. Inoue, H. Tomita, and Y. Ike, Antimicrob. Agents Chemother., 50:1202-1212, 2006). For the first time, the antimicrobial activity spectra for enterocins L50A and L50B were determined separately and included a wide range of gram-positive bacteria but also a few gram-negative strains that were weakly sensitive. Slight differences in the activities of enterocins L50A and L50B were observed, as gram positive bacteria showed an overall higher level of sensitivity to L50A than to L50B, as opposed to gram-negative ones. Conversely, enterocin IT showed a very narrow antimicrobial spectrum that was limited to E. faecium strains, one strain of Bacillus subtilis, and one strain of Lactococcus lactis. This study showed that E. faecium IT62, a grass-borne strain, produces bacteriocins with very different activity features and structures that may be found in strains associated with food or those of clinical origin, which demonstrates that a particular enterocin structure may be widespread and not related to the producer's origin. PMID- 18391038 TI - Bactericidal and sterilizing activities of several orally administered combined regimens against Mycobacterium ulcerans in mice. AB - Treatment with rifampin-clarithromycin or moxifloxacin-clarithromycin for 8 weeks displayed promising bactericidal activity against Mycobacterium ulcerans in mice; none of the mice treated with rifampin-clarithromycin relapsed, whereas 59% of those treated with moxifloxacin-clarithromycin relapsed after treatment was stopped. The bactericidal and sterilizing activities of the five-times-weekly (5/7) administration of 5 mg of rifapentine/kg of body weight, either alone or in combination, were virtually identical to those of the corresponding regimens with 10 mg of rifampin/kg of body weight; however, because of the long half-life of rifapentine, accumulation of the drug after 5/7 administration is a concern. The bactericidal activity of 20 mg/kg rifapentine in monotherapy or 20 mg/kg rifapentine in combination with 150 mg/kg streptomycin or 200 mg/kg moxifloxacin administered twice weekly was as effective as the corresponding regimens containing 10 mg/kg rifampin administered 5/7, suggesting that Buruli ulcer might be treated with intermittently administered rifapentine-containing combinations. PMID- 18391039 TI - Lack of an effect of human immunodeficiency virus coinfection on the pharmacokinetics of entecavir in hepatitis B virus-infected patients. AB - Entecavir is a guanosine nucleoside analogue approved for the treatment of chronic hepatitis B virus (HBV) infection. The impact of human immunodeficiency virus (HIV) coinfection on the pharmacokinetics (PK) of entecavir was examined by nonlinear mixed-effects modeling. Plasma concentration data from HIV- and HBV coinfected patients were analyzed in conjunction with data from HBV-monoinfected patients, and HIV coinfection was tested as a covariate on oral clearance (CL/F). The estimated population averages of intercompartmental clearance and the volumes of distribution in the central and peripheral compartments obtained with a 1-mg dose were 34.2 liters/h (interindividual variability, 30.2%), 115 liters (interindividual variability, 39.2%), and 1,830 liters (interindividual variability, 74%), respectively. CL/F was found to be a function of creatinine clearance, but HIV confection did not show any effect on CL/F. The geometric mean (GM) of individual Bayesian estimates of the steady-state area under the concentration-time curve following 1-mg daily doses were 39.3 and 38.8 ng x h/ml in HIV- and HBV-coinfected and HBV-monoinfected patients, respectively. The adjusted GM ratio (1.01; 90% confidence interval, 0.91 to 1.12) was within the bioequivalence criteria boundary (0.80 to 1.25). In conclusion, the proposed model adequately described the entecavir PK in HBV- and HIV-coinfected patients and HBV-monoinfected patients, and the entecavir exposures were comparable in the two patient populations. PMID- 18391040 TI - Crystal structures of biapenem and tebipenem complexed with penicillin-binding proteins 2X and 1A from Streptococcus pneumoniae. AB - Biapenem is a parenteral carbapenem antibiotic that exhibits wide-ranging antibacterial activity, remarkable chemical stability, and extensive stability against human renal dehydropeptidase-I. Tebipenem is the active form of tebipenem pivoxil, a novel oral carbapenem antibiotic that has a high level of bioavailability in humans, in addition to the above-mentioned features. beta lactam antibiotics, including carbapenems, target penicillin-binding proteins (PBPs), which are membrane-associated enzymes that play essential roles in peptidoglycan biosynthesis. To envisage the binding of carbapenems to PBPs, we determined the crystal structures of the trypsin-digested forms of both PBP 2X and PBP 1A from Streptococcus pneumoniae strain R6, each complexed with biapenem or tebipenem. The structures of the complexes revealed that the carbapenem C-2 side chains form hydrophobic interactions with Trp374 and Thr526 of PBP 2X and with Trp411 and Thr543 of PBP 1A. The Trp and Thr residues are conserved in PBP 2B. These results suggest that interactions between the C-2 side chains of carbapenems and the conserved Trp and Thr residues in PBPs play important roles in the binding of carbapenems to PBPs. PMID- 18391041 TI - Apoptosis induced by aminoglycosides in LLC-PK1 Cells: comparative study of neomycin, gentamicin, amikacin, and isepamicin using electroporation. AB - Levels of apoptosis induction (4',6'-diamidino-2-phenylindole staining, activation of caspase 3) for aminoglycosides were compared by using renal LLC-PK1 cells. Amikacin caused less apoptosis than gentamicin in incubated cells. In electroporated cells, neomycin B and gentamicin caused apoptosis in the 0.03 to 0.1 mM range, isepamicin required larger concentrations (0.2 mM), and amikacin was without effect. PMID- 18391042 TI - Evaluation of the VITEK 2 advanced expert system for reporting piperacillin susceptibility in Klebsiella spp. PMID- 18391043 TI - Effects of mutagenic and chain-terminating nucleotide analogs on enzymes isolated from hepatitis C virus strains of various genotypes. AB - The development of effective therapies for hepatitis C virus (HCV) must take into account genetic variation among HCV strains. Response rates to interferon-based treatments, including the current preferred treatment of pegylated alpha interferon administered with ribavirin, are genotype specific. Of the numerous HCV inhibitors currently in development as antiviral drugs, nucleoside analogs that target the conserved NS5B active site seem to be quite effective against diverse HCV strains. To test this hypothesis, we examined the effects of a panel of nucleotide analogs, including ribavirin triphosphate (RTP) and several chain terminating nucleoside triphosphates, on the activities of purified HCV NS5B polymerases derived from genotype 1a, 1b, and 2a strains. Unlike the genotype specific effects on NS5B activity reported previously for nonnucleoside inhibitors (F. Pauwels, W. Mostmans, L. M. Quirynen, L. van der Helm, C. W. Boutton, A. S. Rueff, E. Cleiren, P. Raboisson, D. Surleraux, O. Nyanguile, and K. A. Simmen, J. Virol. 81:6909-6919, 2007), only minor differences in inhibition were observed among the various genotypes; thus, nucleoside analogs that are current drug candidates may be more promising for treatment of a broader variety of HCV strains. We also examined the effects of RTP on the HCV NS3 helicase/ATPase. As with the polymerase, only minor differences were observed among 1a-, 1b-, and 2a-derived enzymes. RTP did not inhibit the rate of NS3 helicase-catalyzed DNA unwinding but served instead as a substrate to fuel unwinding. NS3 added to RNA synthesis reactions relieved inhibition of the polymerase by RTP, presumably due to RTP hydrolysis. These results suggest that NS3 can limit the incorporation of ribavirin into viral RNA, thus reducing its inhibitory or mutagenic effects. PMID- 18391044 TI - Genetic environment of 16S rRNA methylase gene rmtD. AB - The genetic environment of the 16S rRNA methylase gene rmtD was investigated. rmtD was flanked by a novel ISCR motif located downstream of class I integron In163 in the original Pseudomonas aeruginosa strain. rmtD found in Klebsiella pneumoniae appeared to have been mobilized from P. aeruginosa by an IS26-mediated event. PMID- 18391045 TI - Chronic respiratory infections by mucoid carbapenemase-producing Pseudomonas aeruginosa strains, a new potential public health problem. PMID- 18391046 TI - RNAIII-inhibiting peptide enhances healing of wounds infected with methicillin resistant Staphylococcus aureus. AB - Quorum sensing is a mechanism through which a bacterial population receives input from neighboring cells and elicits an appropriate response to enable survival within the host. Inhibiting quorum sensing by RNAIII-inhibiting peptide (RIP) has been demonstrated as a very effective mode of prevention and therapy for device associated staphylococcal infections and was tested here for healing of wounds that are otherwise resistant to conventional antibiotics. Wounds, established through the panniculus carnosus of BALB/c mice, were inoculated with 5 x 10(7) CFU of methicillin-resistant Staphylococcus aureus. Mice were treated with Allevyn, RIP-soaked Allevyn (containing 20 microg RIP), daily intraperitoneal teicoplanin (7 mg/kg of body weight), Allevyn and teicoplanin, and RIP-soaked Allevyn and daily intraperitoneal teicoplanin. The main outcome measures were quantitative bacterial culture and histological examination with assessment of microvessel density and of vascular endothelial growth factor (VEGF) expression in tissue sections. Treatment with RIP-soaked Allevyn together with teicoplanin injection greatly reduced the bacterial load to 13 CFU/g (control untreated animals had 10(8) CFU/g bacteria). All other treatments were also significantly effective but only reduced the bacterial load to about 10(3) CFU/ml. Histological examination indicated that only treatment with RIP-soaked Allevyn with teicoplanin injection restored epithelial, granulation, and collagen scores, as well as microvessel density and VEGF expression, to the levels found with uninfected mice. In conclusion, we observed that RIP may be useful for the management of infected wounds and that it could represent an exciting and future alternative to the conventional antibiotics, at present considered the gold standard treatments for methicillin-resistant S. aureus infections. PMID- 18391047 TI - Tolevamer, an anionic polymer, neutralizes toxins produced by the BI/027 strains of Clostridium difficile. AB - Clostridium difficile-associated diarrhea (CDAD) is caused by the toxins the organism produces when it overgrows in the colon as a consequence of antibiotic depletion of normal flora. Conventional antibiotic treatment of CDAD increases the likelihood of recurrent disease by again suppressing normal bacterial flora. Tolevamer, a novel toxin-binding polymer, was developed to ameliorate the disease without adversely affecting normal flora. In the current study, tolevamer was tested for its ability to neutralize clostridial toxins produced by the epidemic BI/027 strains, thereby preventing toxin-mediated tissue culture cell rounding. The titers of toxin-containing C. difficile culture supernatants were determined using confluent cell monolayers, and then the supernatants were used in assays containing dilutions of tolevamer to determine the lowest concentration of tolevamer that prevented > or =90% cytotoxicity. Tolevamer neutralized toxins in the supernatants of all C. difficile strains tested. Specific antibodies against the large clostridial toxins TcdA and TcdB also neutralized the cytopathic effect, suggesting that tolevamer is specifically neutralizing these toxins and that the binary toxin (whose genes are carried by the BI/027 strains) is not a significant source of cytopathology against tissue culture cells in vitro. PMID- 18391048 TI - Parenting specificity: an examination of the relation between three parenting behaviors and child problem behaviors in the context of a history of caregiver depression. AB - The aim of this study was to advance our understanding of the relations between three specific parenting behaviors (warmth, monitoring, and discipline) and two child outcomes (internalizing and externalizing problems) within the context of parental depression. Using an approach recommended by A. Caron, B. Weiss, V. Harris, and T. Carron (2006), unique and differential specificity were examined. Ninety-seven parents with a history of depression and 136 of their 9- to 15-year old children served as participants. Children reported parenting behaviors and parents reported child problem behaviors. The findings indicated that warmth/involvement, but not monitoring or discipline, was uniquely related to externalizing problems and differentially related to internalizing and externalizing problems. The findings suggest that parental warmth has implications for interventions conducted with children living in families with a history of parental depression. PMID- 18391049 TI - Improving treatment adherence in bipolar disorder: a review of current psychosocial treatment efficacy and recommendations for future treatment development. AB - Treatment adherence is a frequent problem in bipolar disorder, with research showing that more than 60% of bipolar patients are at least partially nonadherent to medications. Treatment nonadherence is consistently predictive of a number of negative outcomes in bipolar samples, and the discontinuation of mood stabilizers places these patients at high risk for relapse. Several types of adjunctive treatment (family, psychoeducational, cognitive-behavioral) have been investigated for improving symptoms and functioning in bipolar patients with some success. To date, less attention has been paid to developing treatments specifically to promote treatment adherence to and engagement with pharmacological as well as behavioral treatments in patients with bipolar disorder. First, we review the effects of adjunctive interventions specifically on treatment adherence outcomes in 14 published clinical trials. Based on this empirical knowledge base, we present a preliminary description of the treatment strategies that appear most promising for improving adherence. The article also provides research recommendations for developing more effective interventions for the purpose of improving bipolar treatment adherence. Finally, special treatment considerations, including the potential impact of comorbid substance abuse and bipolar depression, are discussed. PMID- 18391050 TI - Distress tolerance treatment for early-lapse smokers: rationale, program description, and preliminary findings. AB - A significant percentage of individuals attempting smoking cessation lapse within a matter of days, and very few are able to recover to achieve long-term abstinence. This observation suggests that many smokers may have quit-attempt histories characterized exclusively by early lapses to smoking following quit attempts. Recent negative-reinforcement conceptualizations of early lapse to smoking suggest that individuals' reactions to withdrawal and inability to tolerate the experience of these symptoms, rather than withdrawal severity itself, may represent an important treatment target in the development of new behavioral interventions for this subpopulation of smokers. This article presents the theoretical rationale and describes a novel, multicomponent distress tolerance treatment for early-lapse smokers that incorporates behavioral and pharmacological elements of standard smoking-cessation treatment, whereas drawing distress-tolerance elements from exposure-based and Acceptance and Commitment Therapy-based treatment approaches. Preliminary data from a pilot study (N = 16) are presented, and clinical implications are discussed. PMID- 18391051 TI - Cognitive-behavioral treatment for panic disorder with agoraphobia: a randomized, controlled trial and cost-effectiveness analysis. AB - A randomized, controlled trial was conducted to examine the cost-effectiveness of cognitive-behavioral treatment (CBT) for panic disorder with agoraphobia. A total of 100 participants were randomly assigned to standard (n = 33), group (n = 35), and brief (n = 32) treatment conditions. Results show significant clinical and statistical improvement on standard symptom measures and quality of life from baseline to posttreatment and 3-month follow-up, with no significant differences between treatment conditions. Compared with standard CBT, brief and group CBT incurred lower treatment costs and had a superior cost-effectiveness ratio, suggesting the potential of these alternative treatment conditions in increasing access to effective treatment. PMID- 18391052 TI - Influence of appearance-related TV commercials on body image state. AB - This study investigates the influence of media exposure on body image state in eating-disordered (ED) patients. The attitudinal and perceptual components of body image are assessed, as well as any associations with dysfunctional cognitions and behavioral consequences. Twenty-five ED patients and 25 non-ED controls (ND) viewed commercials either featuring appearance (AC; 5 min) or not featuring appearance (NC; 5 min). Both perceptual and attitudinal body image components changed markedly after the AC condition for ED patients, compared with the ND group and NC condition. Cognitions referring to dietary restraint and internalization/social comparison also changed significantly in ED patients depending on the experimental manipulation, whereas thoughts about body and self esteem did not. The results suggest that media exposure acts as a stimulus that triggers body-related schemas. Partial support is given to cognitive-behavioral models of eating disorders, which postulate an association between cognitive bias, body image disturbances, and compensatory behavioral consequences. PMID- 18391053 TI - Single-participant assessment of treatment mediators: strategy description and examples from a behavioral activation intervention for depressed adolescents. AB - Determining the means by which effective psychotherapy works is critical. A generally recommended strategy for identifying the potential causal variables is to conduct group-level statistical tests of treatment mediators. Herein the case is made for also assessing mediators of treatment outcome at the level of the individual participant. Single-participant assessment of mediators requires documenting, for each participant, that treatment was received, that change occurred on the mediator and relevant clinical outcome measures, and that the change on the mediator happened at an expected time in the treatment protocol and prior to substantive change on the dependent variable. Data from four depressed adolescents who demonstrated remission following a behavioral activation intervention illustrate the use of the approach in assessing whether changes in activation level or negative thinking mediated the changes in depression. For two participants, increased activation appeared to be a mediator, whereas decreased dysfunctional thinking never emerged as a plausible mediator. It is concluded that single-participant assessment of mediators of treatment outcome offers a useful additional tool for determining possible mechanisms of action in effective psychotherapy. PMID- 18391054 TI - Coping with pain in the motivational context of values: comparison between an acceptance-based and a cognitive control--based protocol. AB - This study compares the effect of an acceptance-based protocol (ACT) and a cognitive control-based (CONT) protocol on three measures of pain coping: tolerance, self-report, and believability. Specific methodological controls were employed to further isolate the role of the value of participating in a pain task, compared to previous investigations on the alteration of the function of aversive stimulation. Twenty participants were randomly assigned to one of the conditions (ACT vs. CONT), and a pre-post design was used. In the ACT condition, the protocol established a relation of coordination between the pain-related thoughts and the actions in the valued direction. In the CONT condition, the protocol established a relation of opposition between the same aspects. Results show an increase in pain tolerance and a reduction of self-reported pain at posttest for both conditions. However, ACT participants showed significantly lower believability of pain than did CONT participants. Conceptual and clinical implications are discussed. PMID- 18391055 TI - How nonrecidivism affects predictive accuracy: evidence from a cross-validation of the Ontario Domestic Assault Risk Assessment (ODARA). AB - Prediction effect sizes such as ROC area are important for demonstrating a risk assessment's generalizability and utility. How a study defines recidivism might affect predictive accuracy. Nonrecidivism is problematic when predicting specialized violence (e.g., domestic violence). The present study cross-validates the ability of the Ontario Domestic Assault Risk Assessment (ODARA) to distinguish subsequent recidivists and nonrecidivists among 391 new cases with less extensive criminal records than previous cross-validation samples, base rate=27%, ROC area=.67. Excluding ambiguous nonrecidivists increases the base rate to 33%, ROC area=.74. Random samples of 50 recidivists and 50 unambiguous nonrecidivists yield ROC areas from .71 to .80. Published norms significantly underestimate official recidivism. Ambiguous nonrecidivism is prevalent and leads to underestimating base rates and predictive accuracy. PMID- 18391056 TI - Violent behavior in female inmates: possible predictors. AB - Research findings have been equivocal regarding the relationship between experiencing trauma and exhibiting violent behavior in women. This study seeks to determine predictors of violent behavior in female inmates utilizing various conceptualizations of traumatic experiences. Results indicate a significant univariate relationship between experiencing more frequent physical abuse experiences, both interpersonal and noninterpersonal types of trauma, and both physical and sexual assaults with frequency of engagement in violent behavior. The regression model explained approximately 21.0% of the variance in violent behavior. Furthermore, frequency of physical abuse experiences was a significant predictor of frequency of engagement in violent behavior and explained 12.74% of unique variance. These results suggest that the frequency of physical abuse experiences may serve as a risk factor for women's violent behavior. Although more research is needed, it appears that addressing the potential for violence in women who experience abuse may be an appropriate target of treatment. PMID- 18391057 TI - Development of a psychometric typology of child molesters: implications for treatment. AB - Cluster analysis of psychometric data measuring offense-specific and socioadequacy problems from a sample of 437 untreated child molesters was carried out. This analysis identified three clusters: Cluster 1 contained men who reported low levels of self-esteem and intimacy and an inability to deal with negative emotions, Cluster 2 contained men who demonstrated a poor understanding of the harm caused to their victims, and Cluster 3 contained offenders who had global offense-specific and socioaffective problems. However, there were no significant differences among the three groups of child molesters in their offense characteristics or their risk levels to support the clustering. It is suggested that these psychological clusters have implications for differing treatment interventions with these three groups. PMID- 18391058 TI - Hate crimes and stigma-related experiences among sexual minority adults in the United States: prevalence estimates from a national probability sample. AB - Using survey responses collected via the Internet from a U.S. national probability sample of gay, lesbian, and bisexual adults (N = 662), this article reports prevalence estimates of criminal victimization and related experiences based on the target's sexual orientation. Approximately 20% of respondents reported having experienced a person or property crime based on their sexual orientation; about half had experienced verbal harassment, and more than 1 in 10 reported having experienced employment or housing discrimination. Gay men were significantly more likely than lesbians or bisexuals to experience violence and property crimes. Employment and housing discrimination were significantly more likely among gay men and lesbians than among bisexual men and women. Implications for future research and policy are discussed. PMID- 18391059 TI - Child maltreatment profiles and adjustment problems in high-risk adolescents. AB - The purpose of this article is to identify profiles of maltreatment experiences in a sample of high-risk adolescents and to investigate the relationship between the derived profiles and psychological adjustment. Participants are 1,131 youth between the ages of 12 and 18 years involved with publicly funded mental health and social services. Information on physical, sexual, and emotional maltreatment and psychological symptoms are obtained in interviews with adolescents and their primary caregivers. Using latent profile analysis, three maltreatment profiles are identified: "sexual+physical+emotional maltreatment," "physical+emotional maltreatment," and "low maltreatment." Adolescents in the two maltreatment profiles generally have significantly higher scores on symptom scales compared with those in the "low maltreatment" profile, but scores in the two maltreatment profiles do not differ. Findings highlight the need for agencies to identify and provide appropriate intervention for youth who experience multiple types of maltreatment. PMID- 18391060 TI - Women arrested for partner violence and substance use: an exploration of discrepancies in the literature. AB - Discrepancies exist in research examining substance problems within groups of women arrested for intimate partner violence (IPV). In some studies women IPV arrestees have been found to be at high risk for substance-related problems, whereas in others they are found to be at low risk for substance-related problems. The current study explores these discrepancies then compares a sample of women court ordered to a domestic violence diversion program (n = 78) to a matched sample of men (n = 78) in the same program using police report information, self-report measures, the Substance Abuse Subtle Screening Inventory 3, and the Millon Clinical Multiaxial Inventory-III alcohol and drug subscales. Women IPV arrestees had a low occurrence of substance use during the arresting incident, a low incidence of substance-related diagnoses, and fewer substance related problems than did men IPV arrestees. However, women demonstrated personality structures susceptible to nondiagnosable, hazardous drinking patterns. PMID- 18391061 TI - Either a Th17 or a Th1 effector response can drive autoimmunity: conditions of disease induction affect dominant effector category. AB - Experimental autoimmune uveitis (EAU) represents autoimmune uveitis in humans. We examined the role of the interleukin (IL)-23-IL-17 and IL-12-T helper cell (Th)1 pathways in the pathogenesis of EAU. IL-23 but not IL-12 was necessary to elicit disease by immunization with the retinal antigen (Ag) interphotoreceptor retinoid binding protein (IRBP) in complete Freund's adjuvant. IL-17 played a dominant role in this model; its neutralization prevented or reversed disease, and Th17 effector cells induced EAU in the absence of interferon (IFN)-gamma. In a transfer model, however, a polarized Th1 line could induce severe EAU independently of host IL-17. Furthermore, induction of EAU with IRBP-pulsed mature dendritic cells required generation of an IFN-gamma-producing effector response, and an IL-17 response by itself was insufficient to elicit pathology. Finally, genetic deficiency of IL-17 did not abrogate EAU susceptibility. Thus, autoimmune pathology can develop in the context of either a Th17 or a Th1 effector response depending on the model. The data suggest that the dominant effector phenotype may be determined at least in part by conditions present during initial exposure to Ag, including the quality/quantity of Toll-like receptor stimulation and/or type of Ag-presenting cells. These data also raise the possibility that the nonredundant requirement for IL-23 in EAU may extend beyond its role in promoting the Th17 effector response and help provide a balance in the current Th1 versus Th17 paradigm. PMID- 18391062 TI - Synthetic chemerin-derived peptides suppress inflammation through ChemR23. AB - Chemerin is a chemotactic protein that binds to the G protein-coupled receptor, ChemR23. We demonstrate that murine chemerin possesses potent antiinflammatory properties that are absolutely dependent on proteolytic processing. A series of peptides was designed, and only those identical to specific C-terminal chemerin sequences exerted antiinflammatory effects at picomolar concentrations in vitro. One of these, chemerin15 (C15; A(140)-A(154)), inhibited macrophage (MPhi) activation to a similar extent as proteolyzed chemerin, but exhibited reduced activity as a MPhi chemoattractant. Intraperitoneal administration of C15 (0.32 ng/kg) to mice before zymosan challenge conferred significant protection against zymosan-induced peritonitis, suppressing neutrophil (63%) and monocyte (62%) recruitment with a concomitant reduction in proinflammatory mediator expression. Importantly, C15 was unable to ameliorate zymosan-induced peritonitis in ChemR23( /-) mice, demonstrating that C15's antiinflammatory effects are entirely ChemR23 dependent. In addition, administration of neutralizing anti-chemerin antibody before zymosan challenge resulted in a significant exacerbation of peritoneal inflammation (up to 170%), suggesting an important endogenous antiinflammatory role for chemerin-derived species. Collectively, these results show that chemerin derived peptides may represent a novel therapeutic strategy for the treatment of inflammatory diseases through ChemR23. PMID- 18391063 TI - The orthopoxvirus type I IFN binding protein is essential for virulence and an effective target for vaccination. AB - Nonliving antiviral vaccines traditionally target proteins expressed at the surface of the virion with the hope of inducing neutralizing antibodies. Orthopoxviruses (OPVs), such as the human smallpox virus and the mouse-equivalent ectromelia virus (ECTV; an agent of mousepox), encode immune response modifiers (IRMs) that can increase virulence by decreasing the host immune response. We show that one of these IRMs, the type I interferon (IFN) binding protein (bp) of ECTV, is essential for ECTV virulence and is a natural target of the antibody response. More strikingly, we demonstrate that immunization with recombinant type I IFN bp protects mice from lethal mousepox. Collectively, our experiments have important implications for our understanding of the role of IRMs in OPV virulence and of type I IFNs in OPV infections. Furthermore, our work provides proof of concept that effective antiviral vaccines can be made to prevent disease by targeting virulence factors as an alternative to the traditional approach that attempts to prevent infection by virus neutralization. PMID- 18391064 TI - Arthritis induced by posttranslationally modified (citrullinated) fibrinogen in DR4-IE transgenic mice. AB - Rheumatoid arthritis (RA) is a common autoimmune disease that afflicts the synovium of diarthrodial joints. The pathogenic mechanisms inciting this disease are not fully characterized, but may involve the loss of tolerance to posttranslationally modified (citrullinated) antigens. We have demonstrated that this modification leads to a selective increase in antigenic peptide affinity for major histocompatibility complex (MHC) class II molecules that carry the RA associated shared epitope, such as HLA-DRB1*0401 (DR4). We describe the induction of arthritis in DR4-IE transgenic (tg) mice with citrullinated fibrinogen, a protein commonly found in inflamed synovial tissue and a frequent target of autoantibodies in RA patients. The disease induced in these mice was characterized by synovial hyperplasia followed by ankylosis, but lacked a conspicuous polymorphonuclear cell infiltrate. Immunological analysis of these mice through T cell epitope scanning and antibody microarray analysis identified a unique profile of citrulline-specific reactivity that was not found in DR4-IE tg mice immunized with unmodified fibrinogen or in wild-type C57BL/6 mice immunized with citrullinated fibrinogen, two conditions where arthritis was not observed. These observations directly implicate citrullinated fibrinogen as arthritogenic in the context of RA-associated MHC class II molecules. PMID- 18391065 TI - Coyotes demonstrate how habitat specialization by individuals of a generalist species can diversify populations in a heterogeneous ecoregion. AB - The tendency for individuals to disperse into habitat similar to their natal habitat has been observed in a wide range of species, although its population genetic consequences have received little study. Such behavior could lead to discrete habitat-specific population subdivisions even in the absence of physical dispersal barriers or habitat gaps. Previous studies of coyotes have supported this hypothesis in a small region of California, but its evolutionary significance ultimately depends on the extent and magnitude of habitat-specific subdivision. Here, we investigated these questions using autosomal, Y chromosome, and mitochondrial markers and >2,000 coyotes from a broad region, including 2 adjacent ecoregions with contrasting levels of habitat heterogeneity--the California Floristic Province (CFP) (heterogeneous landscape) and the Desert Prairie ecoregion (DPE) (homogeneous landscape). Consistent with predictions, we found a close correspondence between population genetic structure and habitat subdivisions throughout the CFP and virtual panmixia over the larger DPE. Conversely, although genetic diversity was similar in these 2 ecoregions overall, it was lower within sites of the CFP, as would be the expected consequence of greater genetic drift within subregions. The magnitude of habitat-specific genetic subdivisions (i.e., genetic distance) in the CFP varied considerably, indicating complexity (e.g., asymmetric gene flow or extinction/recolonization), but, in general, was higher than that due to geographic distance or recent human related barriers. Because habitat-specific structure can enhance a species' adaptive potential and resilience to changing environments, these findings suggest the CFP may constitute an evolutionarily important portion of the range for coyotes and sympatric species exhibiting habitat-specific population structure. PMID- 18391066 TI - Systematic survey for novel types of prokaryotic retroelements based on gene neighborhood and protein architecture. AB - Retroelements, elements encoding reverse transcriptase (RT), are ubiquitous in eukaryotes and have a great influence on the evolution of our genome. Detailed information is available on eukaryotic retroelements; however, prokaryotic retroelements are poorly understood. Recently, new types of eukaryotic retroelements were characterized on the basis of their gene composition and their phylogenetic positions. Here we performed a systematic survey to identify novel types of prokaryotic retroelements by analyzing gene neighborhood and protein architecture. We found novel types of gene combination and examined whether they represent actual retroelements. Five monophyletic groups were identified that were distinct from characterized prokaryotic retroelements, showed specific gene combination, were distributed patchily, and included at least 1 example of recent integration. These results strongly indicated the frequent horizontal transfer of these elements. One group encoded DNA polymerase A. A possible function of DNA polymerase A in the life cycle of retroelements is catalyzing second-strand cDNA synthesis, which is DNA polymerization performed using a DNA template not an RNA template. Another group encoded both bacterial primase and carbon-nitrogen hydrolase. Primase is likely to synthesize primers to initiate reverse transcription. Two other groups also encoded carbon-nitrogen hydrolase as a fusion protein with RT. It is difficult to speculate on the function of hydrolase in the life cycle of retroelements. The last group encoded dual RT proteins, which are likely to form heterodimers during replication. The protein sets of these 5 groups of prokaryotic retroelements were completely different from those of eukaryotic retroelements, indicating that the survival constraints of prokaryotic elements were distinct from those of eukaryotic elements. It is likely that these prokaryotic retroelements are maintained as extrachromosomal DNA or RNA or are accidentally integrated into genomes. Our findings presented the possibility that many types of extrachromosomal prokaryotic retroelements remain to be characterized. In addition, we found 8 RT genes were associated with clustered regularly interspaced short palindrome repeats (CRISPRs) of the CRISPR Cas system. These RT genes are likely to work in immunity against RNA phages via cDNA synthesis. PMID- 18391067 TI - The capsid of the T4 phage superfamily: the evolution, diversity, and structure of some of the most prevalent proteins in the biosphere. AB - The Escherichia coli bacteriophage T4 has served as a classic system in phage biology for more than 60 years. Only recently have phylogenetic analyses and genomic comparisons demonstrated the existence of a large, diverse, and widespread superfamily of T4-like phages in the environment. We report here on the T4-like major capsid protein (MCP) sequences that were obtained by targeted polymerase chain reaction (PCR) of marine environmental samples. This analysis was then expanded to include 1,000 s of new sequences of T4-like capsid genes from the metagenomic data obtained during the Sorcerer II Global Ocean Sampling (GOS) expedition. This data compilation reveals that the diversity of the major and minor capsid proteins from the GOS metagenome follows the same general patterns as the sequences from cultured phage genomes. Interestingly, the new MCP sequences obtained by PCR targeted to MCP sequences in environmental samples are more divergent (deeper branching) than the vast majority of the MCP sequences coming from the other sources. The marine T4-like phage population appears to be largely dominated by the T4-like cyanophages. Using approximately 1,400 T4-like MCP sequences from various sources, we mapped the degree of sequence conservation on a structural model of the T4-like MCP. The results indicate that within the T4 superfamily there are some clear phylogenetic groups with regard to the more conserved and more variable domains of the MCP. Such differences can be correlated with variations in capsid morphology, the arrangement of the MCP lattice, and the presence of different capsid accessory proteins between the subgroups of the T4 superfamily. PMID- 18391068 TI - Richard Gardner: nuclear garbologist. Interview by Ruth Williams. PMID- 18391069 TI - Pushing for answers: is myosin V directly involved in moving mitochondria? AB - In budding yeast, the actin-based class V myosin motors, Myo2 and Myo4, transport virtually all organelles from mother to bud during cell division. Until recently, it appeared that mitochondria may be an exception, with studies showing that the Arp2/3 complex is required for their movement. However, several recent studies have proposed that Myo2 has a direct involvement in mitochondria inheritance. In this issue, Altmann et al. (Altmann, K., M. Frank, D. Neumann, S. Jakobs, and B. Westermann. 2008. J. Cell Biol. 181:119-130) provide the strongest support yet that Myo2 and its associated light chain Mlc1 function directly and significantly in both mitochondria-actin interactions and in the movement of mitochondria from mother to bud. The conflicting functions of Arp 2/3 and Myo2 may be reconciled by the existence of multiple pathways involved in mitochondrial transport. PMID- 18391070 TI - Compensatory role for Pyk2 during angiogenesis in adult mice lacking endothelial cell FAK. AB - Focal adhesion kinase (FAK) plays a critical role during vascular development because knockout of FAK in endothelial cells (ECs) is embryonic lethal. Surprisingly, tamoxifen-inducible conditional knockout of FAK in adult blood vessels (inducible EC-specific FAK knockout [i-EC-FAK-KO]) produces no vascular phenotype, and these animals are capable of developing a robust growth factor induced angiogenic response. Although angiogenesis in wild-type mice is suppressed by pharmacological inhibition of FAK, i-EC-FAK-KO mice are refractory to this treatment, which suggests that adult i-EC-FAK-KO mice develop a compensatory mechanism to bypass the requirement for FAK. Indeed, expression of the FAK-related proline-rich tyrosine kinase 2 (Pyk2) is elevated and phosphorylated in i-EC-FAK-KO blood vessels. In cultured ECs, FAK knockdown leads to increased Pyk2 expression and, surprisingly, FAK kinase inhibition leads to increased Pyk2 phosphorylation. Pyk2 can functionally compensate for the loss of FAK because knockdown or pharmacological inhibition of Pyk2 disrupts angiogenesis in i-EC-FAK-KO mice. These studies reveal the adaptive capacity of ECs to switch to Pyk2-dependent signaling after deletion or kinase inhibition of FAK. PMID- 18391071 TI - The eIF4E RNA regulon promotes the Akt signaling pathway. AB - Eukaryotic initiation factor 4E (eIF4E) promotes cellular proliferation and can rescue cells from apoptotic stimuli such as serum starvation. However, the mechanisms underlying apoptotic rescue are not well understood. In this study, we demonstrate that eIF4E overexpression leads to enhanced survival signaling through Akt and that eIF4E requires Akt1 to rescue serum-deprived fibroblasts. Furthermore, a mutant form of eIF4E (W73A), which is messenger RNA (mRNA) export competent but does not promote translation, rescues cells as readily as wild-type eIF4E. We show that eIF4E mediates Akt activation via up-regulation of Nijmegen breakage syndrome 1 (NBS1), a phosphoinositide-3 kinase-Akt pathway upstream activator. Additionally, eIF4E coordinately up-regulates the expression of downstream effectors of the Akt pathway, thereby amplifying Akt signaling effects. A negative regulator of eIF4E, the promyelocytic leukemia protein (PML), suppresses Akt activation and apoptotic rescue. These PML activities likely arise, at least in part, through its inhibition of eIF4E-mediated NBS1 mRNA export. In summary, eIF4E coordinately regulates gene expression to potentiate Akt activation, an activity required for apoptotic rescue. PMID- 18391072 TI - Transmembrane domain-dependent partitioning of membrane proteins within the endoplasmic reticulum. AB - The length and hydrophobicity of the transmembrane domain (TMD) play an important role in the sorting of membrane proteins within the secretory pathway; however, the relative contributions of protein-protein and protein-lipid interactions to this phenomenon are currently not understood. To investigate the mechanism of TMD dependent sorting, we used the following two C tail-anchored fluorescent proteins (FPs), which differ only in TMD length: FP-17, which is anchored to the endoplasmic reticulum (ER) membrane by 17 uncharged residues, and FP-22, which is driven to the plasma membrane by its 22-residue-long TMD. Before export of FP-22, the two constructs, although freely diffusible, were seen to distribute differently between ER tubules and sheets. Analyses in temperature-blocked cells revealed that FP-17 is excluded from ER exit sites, whereas FP-22 is recruited to them, although it remains freely exchangeable with the surrounding reticulum. Thus, physicochemical features of the TMD influence sorting of membrane proteins both within the ER and at the ER-Golgi boundary by simple receptor-independent mechanisms based on partitioning. PMID- 18391073 TI - The class V myosin motor protein, Myo2, plays a major role in mitochondrial motility in Saccharomyces cerevisiae. AB - The actin cytoskeleton is essential for polarized, bud-directed movement of cellular membranes in Saccharomyces cerevisiae and thus ensures accurate inheritance of organelles during cell division. Also, mitochondrial distribution and inheritance depend on the actin cytoskeleton, though the precise molecular mechanisms are unknown. Here, we establish the class V myosin motor protein, Myo2, as an important mediator of mitochondrial motility in budding yeast. We found that mutants with abnormal expression levels of Myo2 or its associated light chain, Mlc1, exhibit aberrant mitochondrial morphology and loss of mitochondrial DNA. Specific mutations in the globular tail of Myo2 lead to aggregation of mitochondria in the mother cell. Isolated mitochondria lacking functional Myo2 are severely impaired in their capacity to bind to actin filaments in vitro. Time-resolved fluorescence microscopy revealed a block of bud directed anterograde mitochondrial movement in cargo binding-defective myo2 mutant cells. We conclude that Myo2 plays an important and direct role for mitochondrial motility and inheritance in budding yeast. PMID- 18391074 TI - Ras signaling directs endothelial specification of VEGFR2+ vascular progenitor cells. AB - Vascular endothelial growth factor receptor 2 (VEGFR2) transmits signals of crucial importance to vasculogenesis, including proliferation, migration, and differentiation of vascular progenitor cells. Embryonic stem cell-derived VEGFR2(+) mesodermal cells differentiate into mural lineage in the presence of platelet derived growth factor (PDGF)-BB or serum but into endothelial lineage in response to VEGF-A. We found that inhibition of H-Ras function by a farnesyltransferase inhibitor or a knockdown technique results in selective suppression of VEGF-A-induced endothelial specification. Experiments with ex vivo whole-embryo culture as well as analysis of H-ras(-/-) mice also supported this conclusion. Furthermore, expression of a constitutively active H-Ras[G12V] in VEGFR2(+) progenitor cells resulted in endothelial differentiation through the extracellular signal-related kinase (Erk) pathway. Both VEGF-A and PDGF-BB activated Ras in VEGFR2(+) progenitor cells 5 min after treatment. However, VEGF A, but not PDGF-BB, activated Ras 6-9 h after treatment, preceding the induction of endothelial markers. VEGF-A thus activates temporally distinct Ras-Erk signaling to direct endothelial specification of VEGFR2(+) vascular progenitor cells. PMID- 18391075 TI - straightjacket is required for the synaptic stabilization of cacophony, a voltage gated calcium channel alpha1 subunit. AB - In a screen to identify genes involved in synaptic function, we isolated mutations in Drosophila melanogaster straightjacket (stj), an alpha(2)delta subunit of the voltage-gated calcium channel. stj mutant photoreceptors develop normal synaptic connections but display reduced "on-off" transients in electroretinogram recordings, indicating a failure to evoke postsynaptic responses and, thus, a defect in neurotransmission. stj is expressed in neurons but excluded from glia. Mutants exhibit endogenous seizure-like activity, indicating altered neuronal excitability. However, at the synaptic level, stj larval neuromuscular junctions exhibit approximately fourfold reduction in synaptic release compared with controls stemming from a reduced release probability at these synapses. These defects likely stem from destabilization of Cacophony (Cac), the primary presynaptic alpha(1) subunit in D. melanogaster. Interestingly, neuronal overexpression of cac partially rescues the viability and physiological defects in stj mutants, indicating a role for the alpha(2)delta Ca(2+) channel subunit in mediating the proper localization of an alpha(1) subunit at synapses. PMID- 18391076 TI - Translocations targeting CCND2, CCND3, and MYCN do occur in t(11;14)-negative mantle cell lymphomas. AB - The genetics of t(11;14)(q13;q32)/cyclin D1-negative mantle cell lymphoma (MCL) is poorly understood. We report here 8 MCL cases lacking t(11;14) or variant CCND1 rearrangement that showed expression of cyclin D1 (2 cases), D2 (2 cases), and D3 (3 cases). One case was cyclin D negative. Cytogenetics and fluorescence in situ hybridization detected t(2;12)(p11;p13)/IGK-CCND2 in one of the cyclin D2 positive cases and t(6;14)(p21;q32)/IGH-CCND3 in one of the cyclin D3-positive cases. Moreover, we identified a novel cryptic t(2;14)(p24;q32) targeting MYCN in 2 blastoid MCLs: one negative for cyclin D and one expressing cyclin D3. Interestingly, both cases showed expression of cyclin E. Notably, all 3 blastoid MCLs showed a monoallelic deletion of RB1 associated with a lack of expression of RB1 protein and monoallelic loss of p16. In sum-mary, this study confirms frequent aberrant expression of cyclin D2 and D3 in t(11;14)-negative MCLs and shows a t(11;14)-independent expression of cy-clin D1 in 25% of present cases. Novel findings include cyclin E expression in 2 t(11;14)-negative MCLs characterized by a cryptic t(2;14)(p24;q32) and identification of MYCN as a new lymphoma oncogene associated with a blastoid MCL. Clinically important is a predisposition of t(11;14)-negative MCLs to the central nervous system involvement. PMID- 18391079 TI - Osteopathic medical education in 2008: course corrections and new horizons. PMID- 18391077 TI - Genotype-phenotype correlation in combined deficiency of factor V and factor VIII. AB - Combined deficiency of factor V and factor VIII (F5F8D) is caused by mutations in one of 2 genes, either LMAN1 or MCFD2. Here we report the identification of mutations for 11 additional F5F8D families, including 4 novel mutations, 2 in MCFD2 and 2 in LMAN1. We show that a novel MCFD2 missense mutation identified here (D81Y) and 2 previously reported mutations (D89A and D122V) abolish MCFD2 binding to LMAN1. Measurement of platelet factor V (FV) levels in 7 F5F8D patients (4 with LMAN1 and 3 with MCFD2 mutations) demonstrated similar reductions to those observed for plasma FV. Combining the current data together with all previous published reports, we performed a genotype-phenotype analysis comparing patients with MCFD2 mutations with those with LMAN1 mutations. A previously unappreciated difference is observed between these 2 classes of patients in the distribution of plasma levels for FV and factor VIII (FVIII). Although there is considerable overlap, the mean levels of plasma FV and FVIII in patients with MCFD2 mutations are significantly lower than the corresponding levels in patients with LMAN1 mutations. No differences in distribution of factor levels are observed by sex. These data suggest that MCFD2 may play a primary role in the export of FV and FVIII from the ER, with the impact of LMAN1 mediated indirectly through its interaction with MCFD2. PMID- 18391078 TI - Critical but divergent roles for CD62L and CD44 in directing blood monocyte trafficking in vivo during inflammation. AB - Using noninvasive in vivo imaging and experimental autoimmune uveoretinitis as a model, we show for the first time that the mechanisms controlling blood monocyte recirculation through peripheral and lymphoid tissues alter during inflammation. The recirculation of monocytes in mice with ocular inflammation but not controls was found to depend on the selectin CD62-ligand (CD62L) and on CD44. Not only was rolling efficiency ablated or markedly reduced in antibody-treated mice, but most of the labeled monocytes also disappeared from the circulation within seconds, anti-CD44-treated monocytes homing to the lymph nodes and anti-CD62L-treated monocytes homing to the spleen. Our data indicate that, although PSGL-1 has a partial role in the transmigration of monocytes into the inflamed retina, CD62L has a key role in regulating recruitment of monocytes to lymphoid tissue from the blood during inflammation and that CD44 is required to maintain CD62L(+) inflammatory monocytes within the circulation during inflammation. This effect was systemic, because sequestered monocytes accumulated in mesenteric as well as draining cervical lymph nodes, and inflammation dependent, because depletion of circulating blood monocytes was much reduced or absent in normal mice and accumulations of adoptively transferred monocytes in the lymphoid tissues did not occur. PMID- 18391080 TI - Pushing bodies through COMs. PMID- 18391082 TI - Medical education summits: building a solid foundation for the future of the osteopathic medical profession. AB - In January 2006, the inaugural Osteopathic Heritage Foundation Medical Education Summit was held to address key issues confronting the osteopathic medical profession, particularly the physician workforce, student recruitment, and the funding of postdoctoral programs. Building off of the draft statements from the first summit, the Medical Education Summit II addressed issues specifically related to osteopathic graduate medical education. The authors provide a brief outline of the summit process, describe the outcomes from both meetings, and discuss the future goals of osteopathic medical education. PMID- 18391083 TI - AACOM projections for growth through 2012: results of a 2007 survey of US Colleges of Osteopathic Medicine. AB - In fall 2006, the American Association of Colleges of Osteopathic Medicine (AACOM) sent an electronic survey to the deans of all 23 colleges of osteopathic medicine (COMs) in the United States. A response rate of 100% was achieved and data were reported to participants in May 2007. In fall 2007, AACOM sent shortened, personalized follow-up surveys to the same sample group, asking participants to confirm or correct their previous responses. The 2007 AACOM Survey of Osteopathic College Growth Plans and Issues took approximately 30 minutes to complete. A response rate of 96% was achieved. The results of this 2007 survey are summarized in light of data in AACOM's forthcoming 2007 Annual Statistical Report on Osteopathic Medical Education. By the 2012-2013 academic year, first-year student enrollment is expected to increase approximately 23%. By 2016-2017, more than 5000 DOs will graduate from COMs annually-a 62% increase over current numbers. When student recruiting efforts focus on demographic and geographic factors, the nation's COMs are most inclined to use these variables to address the needs of underserved rural populations. The majority of participants, regardless of their current plans for institutional expansion, indicated that the availability of clinical training sites is a cause of concern. Survey respondents also commented on the fiscal challenges of expansion. PMID- 18391085 TI - Resistant hypertension: diagnosis, evaluation, and treatment. A scientific statement from the American Heart Association Professional Education Committee of the Council for High Blood Pressure Research. AB - Resistant hypertension is a common clinical problem faced by both primary care clinicians and specialists. While the exact prevalence of resistant hypertension is unknown, clinical trials suggest that it is not rare, involving perhaps 20% to 30% of study participants. As older age and obesity are 2 of the strongest risk factors for uncontrolled hypertension, the incidence of resistant hypertension will likely increase as the population becomes more elderly and heavier. The prognosis of resistant hypertension is unknown, but cardiovascular risk is undoubtedly increased as patients often have a history of long-standing, severe hypertension complicated by multiple other cardiovascular risk factors such as obesity, sleep apnea, diabetes, and chronic kidney disease. The diagnosis of resistant hypertension requires use of good blood pressure technique to confirm persistently elevated blood pressure levels. Pseudoresistance, including lack of blood pressure control secondary to poor medication adherence or white coat hypertension, must be excluded. Resistant hypertension is almost always multifactorial in etiology. Successful treatment requires identification and reversal of lifestyle factors contributing to treatment resistance; diagnosis and appropriate treatment of secondary causes of hypertension; and use of effective multidrug regimens. As a subgroup, patients with resistant hypertension have not been widely studied. Observational assessments have allowed for identification of demographic and lifestyle characteristics associated with resistant hypertension, and the role of secondary causes of hypertension in promoting treatment resistance is well documented; however, identification of broader mechanisms of treatment resistance is lacking. In particular, attempts to elucidate potential genetic causes of resistant hypertension have been limited. Recommendations for the pharmacological treatment of resistant hypertension remain largely empiric due to the lack of systematic assessments of 3 or 4 drug combinations. Studies of resistant hypertension are limited by the high cardiovascular risk of patients within this subgroup, which generally precludes safe withdrawal of medications; the presence of multiple disease processes (eg, sleep apnea, diabetes, chronic kidney disease, atherosclerotic disease) and their associated medical therapies, which confound interpretation of study results; and the difficulty in enrolling large numbers of study participants. Expanding our understanding of the causes of resistant hypertension and thereby potentially allowing for more effective prevention and/or treatment will be essential to improve the long-term clinical management of this disorder. PMID- 18391086 TI - Hypertension: pathways to success. PMID- 18391087 TI - Birth weight and retinal vascular changes. PMID- 18391088 TI - Angiotensin II-induced proliferation of neonatal and adult rat cardiac fibroblasts. PMID- 18391089 TI - Extracellular adenosine attenuates left ventricular hypertrophy through its impact on the protein kinase and phosphatase interaction. PMID- 18391090 TI - Questions regarding the aortic measurements of Mitchell et al. PMID- 18391091 TI - Nitric oxide deficiency and increased adenosine response of afferent arterioles in hydronephrotic mice with hypertension. AB - Afferent arterioles were used to investigate the role of adenosine, angiotensin II, NO, and reactive oxygen species in the pathogenesis of increased tubuloglomerular feedback response in hydronephrosis. Hydronephrosis was induced in wild-type mice, superoxide dismutase-1 overexpressed mice (superoxide dismutase-1 transgenic), and deficient mice (superoxide dismutase-1 knockout). Isotonic contractions in isolated perfused arterioles and mRNA expression of NO synthase isoforms, adenosine, and angiotensin II receptors were measured. In wild type mice, N(G)-nitro-L-arginine methyl ester (L-NAME) did not change the basal arteriolar diameter of hydronephrotic kidneys (-6%) but reduced it in control ( 12%) and contralateral arterioles (-43%). Angiotensin II mediated a weaker maximum contraction of hydronephrotic arterioles (-18%) than in control (-42%) and contralateral arterioles (-49%). The maximum adenosine-induced constriction was stronger in hydronephrotic (-19%) compared with control (-8%) and contralateral kidneys (+/-0%). The response to angiotensin II became stronger in the presence of adenosine in hydronephrotic kidneys and attenuated in contralateral arterioles. L-NAME increased angiotensin II responses of all of the groups but less in hydronephrotic kidneys. The mRNA expression of endothelial NO synthase and inducible NO synthase was upregulated in the hydronephrotic arterioles. No differences were found for adenosine or angiotensin II receptors. In superoxide dismutase-1 transgenic mice, strong but similar L-NAME response ( 40%) was observed for all of the groups. This response was totally abolished in arterioles of hydronephrotic superoxide dismutase-1 knockout mice. In conclusion, hydronephrosis is associated with changes in the arteriolar reactivity of both hydronephrotic and contralateral kidneys. Increased oxidative stress, reduced NO availability, and stronger reactivity to adenosine of the hydronephrotic kidney may contribute to the enhanced tubuloglomerular feedback responsiveness in hydronephrosis and be involved in the development of hypertension. PMID- 18391092 TI - Renin inhibition by aliskiren prevents atherosclerosis progression: comparison with irbesartan, atenolol, and amlodipine. AB - Hypertension is associated with increased risk of cardiovascular diseases. Antihypertensive treatment, particularly blockade of the renin-angiotensin system, contributes to prevent atherosclerosis-mediated cardiovascular events. Direct comparison of different antihypertensive treatments on atherosclerosis and particularly plaque stabilization is sparse. ApoE(-/-) mice with vulnerable (2 kidney, 1-clip renovascular hypertension model) or stable (1-kidney, 1-clip renovascular hypertension model) atherosclerotic plaques were used. Mice were treated with aliskiren (renin inhibitor), irbesartan (angiotensin-receptor blocker), atenolol (beta-blocker), or amlodipine (calcium channel blocker). Atherosclerosis characteristics were assessed. Hemodynamic and hormonal parameters were measured. Aliskiren and irbesartan significantly prevented atherosclerosis progression in 2-kidney, 1-clip mice. Indeed, compared with untreated animals, plaques showed thinner fibrous cap (P<0.05); smaller lipid core (P<0.05); decreased media degeneration, layering, and macrophage content (P<0.05); and increased smooth muscle cell content (P<0.05). Interestingly, aliskiren significantly increased the smooth muscle cell compared with irbesartan. Despite similar blood pressure lowering, only partial plaque stabilization was attained by atenolol and amlodipine. Amlodipine increased plaque smooth muscle cell content (P<0.05), whereas atenolol decreased plaque inflammation (P<0.05). This divergent effect was also observed in 1-kidney, 1 clip mice. Normalizing blood pressure by irbesartan increased the plasma renin concentration (5932+/-1512 ng/mL per hour) more than normalizing it by aliskiren (16085+/-5628 ng/mL per hour). Specific renin-angiotensin system blockade prevents atherosclerosis progression. First, evidence is provided that direct renin inhibition mediates atherosclerotic plaque stabilization. In contrast, beta blocker and calcium channel blocker treatment only partially stabilize plaques differently influencing atherogenesis. Angiotensin II decisively mediates plaque vulnerability. The plasma renin concentration measurement by an indirect method did not confirm the excessive increase of plasma renin concentration reported in the literature during aliskiren compared with irbesartan or amlodipine treatment. PMID- 18391094 TI - Atrial fibrillation and isolated systolic hypertension: the systolic hypertension in the elderly program and systolic hypertension in the elderly program-extension study. AB - We performed a post hoc analysis of the Systolic Hypertension in the Elderly Program database to assess the incidence of atrial fibrillation in the elderly hypertensive population, its influence on cardiovascular events, and whether antihypertensive treatment can prevent its onset. The Systolic Hypertension in the Elderly Program was a double-blind placebo-controlled trial in 4736 subjects with isolated systolic hypertension aged >or=60 years. Atrial fibrillation was an exclusion criterion from the trial. Participants were randomly assigned to stepped care treatment with chlorthalidone and atenolol (n=2365) or placebo (n=2371). The occurrence of atrial fibrillation and cardiovascular events over 4.7 years as well as the determination of cause of death at 4.7 and 14.3 years were followed. Ninety-eight subjects (2.06%) developed atrial fibrillation over 4.7 years mean follow-up, without significant difference between treated and placebo groups. Atrial fibrillation increased the risk for: total cardiovascular events (RR 1.69; 95% CI 1.21 to 2.36), rapid death (RR 3.29; 95% CI 1.08 to 10.00), total (RR 5.10; 95% CI 3.12 to 8.37) and nonfatal left ventricular failure (RR 5.31; 95% CI 3.09 to 9.13). All-cause and total cardiovascular death were significantly increased in the atrial fibrillation group at 4.7 years (HR 3.44; 95% CI 2.18 to 5.42; HR 2.39; 95% CI 1.05 to 5.43) and 14.3 years follow-up (HR 2.33; 95% CI 1.83 to 2.98; HR 2.21; 95% CI 1.54 to 3.17). Atrial fibrillation increased the risk for total cardiovascular events, rapid death, and left ventricular failure. All-cause mortality and total cardiovascular mortality were significantly increased in hypertensives with atrial fibrillation at 4.7 and 14.3 years follow-up. PMID- 18391093 TI - Ecto-5'-nucleotidase deficiency exacerbates pressure-overload-induced left ventricular hypertrophy and dysfunction. AB - This study examined whether endogenous extracellular adenosine acts to facilitate the adaptive response of the heart to chronic systolic overload. To examine whether endogenous extracellular adenosine can protect the heart against pressure overload-induced heart failure, transverse aortic constriction was performed on mice deficient in extracellular adenosine production as the result of genetic deletion of CD73. Although there was no difference in left ventricular size or function between CD73-deficient mice (knockout [KO] mice) and wild-type mice under unstressed conditions, aortic constriction for 2 or 4 weeks induced significantly more myocardial hypertrophy, left ventricular dilation, and left ventricular dysfunction in KO mice compared with wild-type mice. Thus, after 2 weeks of transverse aortic constriction, left ventricular fractional shortening decreased to 27.4+/-2.5% and 21.9+/-1.7% in wild-type and KO mice, respectively (P<0.05). Consistent with a role of adenosine in reducing tissue remodeling, KO mice displayed increased myocardial fibrosis and myocyte hypertrophy compared with wild-type mice. Furthermore, adenosine treatment reduced phenylephrine induced cardiac myocyte hypertrophy and collagen production in cultured neonatal rat cardiac myocytes and cardiac fibroblasts, respectively. Consistent with a role for adenosine in modulating cardiomyocyte hypertrophy, KO mice demonstrated increased activation of mammalian target of rapamycin signaling, accompanied by higher expression of the hypertrophy marker atrial natriuretic peptide. Conversely, the adenosine analogue 2-chloro-adenosine significantly reduced cell size, mammalian target of rapamycin/p70 ribosomal S6 kinase activation, and atrial natriuretic peptide expression in cultured neonatal cardiomyocytes. These data demonstrate that CD73 helps to preserve cardiac function during chronic systolic overload by preventing maladaptive tissue remodeling. PMID- 18391095 TI - Wave reflection in systolic hypertension: smaller stature, shorter aorta: higher pulse pressure? PMID- 18391096 TI - Deficiency of bradykinin receptor B2 is not detrimental in experimental stroke. PMID- 18391097 TI - Structure-based identification of small-molecule angiotensin-converting enzyme 2 activators as novel antihypertensive agents. AB - Angiotensin-converting enzyme 2 (ACE2) is a key renin-angiotensin system enzyme involved in balancing the adverse effects of angiotensin II on the cardiovascular system, and its overexpression by gene transfer is beneficial in cardiovascular disease. Therefore, our objectives were 2-fold: to identify compounds that enhance ACE2 activity using a novel conformation-based rational drug discovery strategy and to evaluate whether such compounds reverse hypertension-induced pathophysiologies. We used a unique virtual screening approach. In vitro assays revealed 2 compounds (a xanthenone and resorcinolnaphthalein) that enhanced ACE2 activity in a dose-dependent manner. Acute in vivo administration of the xanthenone resulted in a dose-dependent transient and robust decrease in blood pressure (at 10 mg/kg, spontaneously hypertensive rats decreased 71+/-9 mm Hg and Wistar-Kyoto rats decreased 21+/-8 mm Hg; P<0.05). Chronic infusion of the xanthenone (120 microg/day) resulted in a modest decrease in the spontaneously hypertensive rat blood pressure (17 mm Hg; 2-way ANOVA; P<0.05), whereas it had no effect in Wistar-Kyoto rats. Strikingly, the decrease in blood pressure was also associated with improvements in cardiac function and reversal of myocardial, perivascular, and renal fibrosis in the spontaneously hypertensive rats. We conclude that structure-based screening can help identify compounds that activate ACE2, decrease blood pressure, and reverse tissue remodeling. Administration of ACE2 activators may be a valid strategy for antihypertensive therapy. PMID- 18391098 TI - Salutary effect of kallistatin in salt-induced renal injury, inflammation, and fibrosis via antioxidative stress. AB - An inverse relationship exists between kallistatin levels and salt-induced oxidative stress in Dahl-salt sensitive rats. We further investigated the role of kallistatin in inhibiting inflammation and fibrosis through antioxidative stress in Dahl-salt sensitive rats and cultured renal cells. High-salt intake in Dahl salt sensitive rats induced elevation of thiobarbituric acid reactive substances (an indicator of lipid peroxidation), malondialdehyde levels, reduced nicotinamide-adenine dinucleotide phosphate oxidase activity, and superoxide formation, whereas kallistatin gene delivery significantly reduced these oxidative stress parameters. Kallistatin treatment improved renal function and reduced kidney damage as evidenced by diminished proteinuria and serum urea nitrogen levels, glomerular sclerosis, tubular damage, and protein cast formation. Kallistatin significantly decreased interstitial monocyte-macrophage infiltration and the expression of tumor necrosis factor-alpha, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1. Kallistain also reduced collagen fraction volume and the deposition and expression of collagen types I and III. Renal protection by kallistatin was associated with increased NO levels and endothelial NO synthase expression and decreased p38 mitogen-activated protein kinase, extracellular signal-regulated kinase phosphorylation, and transforming growth factor-beta1 expression. Moreover, kallistatin attenuated tumor necrosis factor-alpha-induced intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression via inhibition of reactive oxygen species formation and p38 mitogen-activated protein kinase and nuclear factor kappaB activation in cultured proximal tubular cells. Kallistatin inhibited fibronectin and collagen expression by suppressing angiotensin II-induced reactive oxygen species generation and transforming growth factor-beta1 expression in cultured mesangial cells. These combined findings reveal that kallistatin is a novel antioxidant, which prevents salt-induced kidney injury, inflammation, and fibrosis by inhibiting reactive oxygen species-induced proinflammatory cytokine and transforming growth factor-beta1 expression. PMID- 18391099 TI - Collecting duct-derived endothelin regulates arterial pressure and Na excretion via nitric oxide. AB - Mice with a collecting duct-specific deletion of endothelin-1 are hypertensive and have impaired Na excretion. Because endothelin-1 activates NO synthase (NOS) in the collecting duct, we hypothesized that impaired renal NO production in knockout mice exacerbates the hypertensive state. Control and knockout mice were treated chronically with N(G)-nitro-l-arginine methyl ester, and blood pressure (BP) and urinary nitrate/nitrite excretion were assessed. On a normal Na diet, knockout systolic BP was 18 mm Hg greater than in controls. N(G)-nitro-l-arginine methyl ester increased BP in control mice by 30 mm Hg and 10 mm Hg in collecting duct-specific deletion of endothelin-1 knockout mice, thereby abolishing the difference in systolic BP between the groups. A high-Na diet increased BP similarly in both groups. Urinary nitrate/nitrite excretion was lower in knockout mice than in controls on normal or high Na intake. In separate experiments, renal perfusion pressure was adjusted in anesthetized mice, and urinary nitrate/nitrite and Na excretion were determined. Similar elevations of BP increased urinary Na and nitrate/nitrite excretion in control mice but to a significantly lesser extent in knockout mice. Isoform-specific NOS activity and expression were determined in renal inner medulla homogenates from control and knockout mice. NOS1 and NOS3 activities were lower in knockout than in control mice given normal or high-Na diets. However, NOS1 or NOS3 protein expressions were similar in both groups on normal or high-Na intake. These data demonstrate that collecting duct derived endothelin-1 is important in the following: (1) chronic N(G)-nitro-l arginine methyl ester-induced hypertension; (2) full expression of pressure dependent changes in sodium excretion; and (3) control of inner medullary NOS1 and NOS3 activity. PMID- 18391100 TI - Menopausal complaints are associated with cardiovascular risk factors. AB - It has been hypothesized that women with vasomotor symptoms differ from those without with respect to cardiovascular risk factors or responses to exogenous hormone therapy. We studied whether the presence and extent of menopausal complaints are associated with cardiovascular risk profile. Data were used from a population-based sample of 5523 women, aged 46 to 57 years, enrolled between 1994 and 1995. Data on menopausal complaints and potential confounders were collected by questionnaires. Total cholesterol, systolic and diastolic blood pressures, and body mass index were measured. Linear and logistic regression analyses were used to analyze the data. Night sweats were reported by 38% and flushing by 39% of women. After multivariate adjustment, women with complaints of flushing had a 0.27-mmol/L (95% CI: 0.15 to 0.39) higher cholesterol level, a 0.60-kg/m(2) (95% CI: 0.35 to 0.84) higher BMI, a 1.59-mm Hg (95% CI: 0.52 to 2.67) higher systolic blood pressure, and a 1.09-mm Hg (95% CI: 0.48 to 1.69) higher diastolic blood pressure compared with asymptomatic women. Flushing was also associated with hypercholesterolemia (odds ratio: 1.52; 95% CI: 1.25 to 1.84) and hypertension (OR: 1.20; 95% CI: 1.07 to 1.34). Results were similar for complaints of night sweating. The findings support the view that menopausal complaints are associated with a less favorable cardiovascular risk profile. These findings substantiate the view that differences in the presence of menopausal symptoms as a reason for using hormone therapy could explain discrepant findings between observational research and trials. PMID- 18391101 TI - A single nucleotide polymorphism in the CYP4F2 but not CYP4A11 gene is associated with increased 20-HETE excretion and blood pressure. AB - Arachidonic acid is a major fatty acid that can be metabolized by the cytochrome P450 enzyme to a number of bioactive eicosanoids. A major metabolite of this oxidation is 20-hydroxyeicosatetraenoic acid, which acts as a potent vasoconstrictor. However, in the kidney, its vasoconstrictor actions can be offset by its natriuretic properties. A guanine-to-adenine polymorphism in the CYP4F2 gene was associated with a reduction in 20-hydroxyeicosatetraenoic acid production in vitro. A thymidine-to-cytosine polymorphism in the CYP4A11 gene reduced catalytic activity by >50% in vitro and was associated with hypertension. The aim was to determine whether these 2 mutations are associated with urinary 20 hydroxyeicosatetraenoic acid excretion and blood pressure in humans. For the CYP4F2, 51% were homozygous for the G allele, 40% were carriers, and 9% were homozygous for the A allele. For CYP4A11, 72% were homozygous for the T allele, 25% were carriers, and 3% were homozygous for the C allele. The CYP4F2 GA/AA genotype was significantly associated with an increase in both 20 hydroxyeicosatetraenoic acid excretion and systolic blood pressure. The CYP4A11 CC/TC genotype was significantly associated with a reduction in 20 hydroxyeicosatetraenoic acid excretion but was not associated with blood pressure. We have demonstrated for the first time in humans that polymorphisms of the CYP4F2 and CYP4A11 genes have opposite effects on 20-hydroxyeicosatetraenoic acid excretion. The positive association between the CYP4F2 GA/AA genotype and both systolic blood pressure and 20-hydroxyeicosatetraenoic acid excretion strengthens a role for 20-hydroxyeicosatetraenoic acid in the modulation of blood pressure. PMID- 18391102 TI - Acute effects of passive smoking on peripheral vascular function. AB - Environmental tobacco smoke (ETS) acutely affects peripheral and coronary vascular tone. Whether ETS exerts specific deleterious effects on aortic wave reflection through nicotine exposure, whether they persist after ETS cessation, and whether the smoke environment impairs microvascular function and increases asymmetrical dimethyl-arginine levels are not known. We tested these hypotheses in a randomized, crossover study design in 11 healthy male nonsmokers. The effects of 1 hour of exposure to ETS, as compared with a nontobacco smoke and normal air, on augmentation index corrected for heart rate and skin microvascular hyperemia to local heating were examined. Augmentation index increased both during (P=0.01) and after (P<0.01) the ETS session but remained unchanged in the nontobacco smoke session when compared with normal air. Nicotine levels after the exposure were related to the peak rise in augmentation index (r=0.84; P<0.01), denoting a predominant role of nicotine in ETS vascular effects. This was confirmed in a second set of experiments (n=14), where the sublingual administration of nicotine was associated with an acute impairment in wave reflection as compared with placebo (P=0.001). Both ETS and nontobacco smokes increased plasma asymmetrical dimethyl-arginine levels (P<0.001), but only ETS reduced the late rise in skin blood flow in response to heating (P=0.03). In conclusion, passive smoking specifically increases aortic wave reflection through a nicotine-dependent pathway and impairs microvascular function, even after the end of the exposure. However, both tobacco and nontobacco passive smoking inhalation increase plasma asymmetrical dimethyl-arginine levels. PMID- 18391103 TI - Kidney dysfunction and sudden cardiac death among women with coronary heart disease. AB - We evaluated the association between kidney dysfunction and sudden cardiac death risk among ambulatory women with coronary heart disease. The Heart and Estrogen Replacement Study evaluated the effects of hormone treatment on cardiovascular events among 2763 postmenopausal women with coronary heart disease. Kidney dysfunction was categorized by estimated glomerular filtration rate (eGFR) using the Modification of Diet in Renal Disease equation. Multivariate proportional hazards models were used to adjust for cardiovascular risk factors, congestive heart failure, and myocardial infarction. At baseline, 37% (n=1027) had an eGFR of >60 mL/min, 54% (n=1503) had an eGFR of 40 to 60 mL/min, and 8% (n=230) had an eGFR of <40 mL/min. During the 6.8-year follow-up period, there were 136 adjudicated sudden cardiac deaths. The rate of sudden cardiac death was higher in those with lower kidney function (0.5% per year among those with an eGFR >60; 0.6% per year with an eGFR between 40 and 60; and 1.7% per year with an eGFR <40 mL/min; P for trend <0.001). After multivariate analysis with baseline risk factors, eGFR at 40 to 60 mL/min was not a significant predictor, but eGFR at <40 mL/min remained strongly associated with sudden cardiac death (hazard ratio: 3.2; 95% CI: 1.9 to 5.3); adjustment for incident congestive heart failure and myocardial infarction during follow-up diminished this association (hazard ratio: 2.3; 95% CI: 1.3 to 3.9), suggesting that congestive heart failure and myocardial infarction mediated only part of the association between kidney dysfunction and sudden cardiac death. Advanced kidney dysfunction is an independent predictor of sudden cardiac death among women with coronary heart disease. PMID- 18391104 TI - H2O2 stimulation of the Cl-/HCO3- exchanger by angiotensin II and angiotensin II type 1 receptor distribution in membrane microdomains. AB - The present study tested the hypothesis that angiotensin II (Ang II)-induced oxidative stress and Ang II-stimulated Cl(-)/HCO(3)(-) exchanger are increased and related to the differential membrane Ang II type 1 (AT(1)) receptor and reduced nicotinamide-adenine dinucleotide phosphate oxidase expression in immortalized renal proximal tubular epithelial (PTE) cells from the spontaneously hypertensive rat (SHR) relative to its normotensive control (Wistar Kyoto rat [WKY]). The exposure of cells to Ang II increased Cl(-)/HCO(3)(-) exchanger activity with EC(50)s of 0.10 and 12.2 nmol/L in SHR and WKY PTE cells, respectively. SHR PTE cells were found to overexpress nicotinamide-adenine dinucleotide phosphate oxidase 2 and 4 and were endowed with an enhanced ability to generate H(2)O(2). The reduced nicotinamide-adenine dinucleotide phosphate oxidase inhibitor apocynin reduced the production of H(2)O(2) in SHR PTE cells and abolished their hypersensitivity to Ang II. The expression of the glycosylated form of the AT(1) receptor in both lipid and nonlipid rafts were higher in SHR cells than in WKY PTE cells. Pretreatment with apocynin reduced the abundance of AT(1) receptors in both microdomains, mainly the glycosylated form of the AT(1) receptor in lipid rafts, in SHR cells but not in WKY PTE cells. In conclusion, differences between WKY and SHR PTE cells in their sensitivity to Ang II correlate with the higher H(2)O(2) generation that provokes an enhanced expression of glycosylated and nonglycosylated AT(1) receptor forms in lipid rafts. PMID- 18391105 TI - Involvement of tumor necrosis factor-alpha in angiotensin II-mediated effects on salt appetite, hypertension, and cardiac hypertrophy. AB - Hypertension is considered a low-grade inflammatory condition induced by various proinflammatory cytokines, including tumor necrosis factor (TNF)-alpha. Recent studies have implicated an involvement of TNF-alpha in the development of salt sensitive hypertension induced by angiotensin II (Ang II). To understand further the relationship between TNF-alpha and Ang II, we examined the responses to Ang II in TNF-alpha knockout (TNF-alpha(-/-)) mice in the present study. A continuous infusion of Ang II (1 microg/kg per minute) for 2 weeks was given to both TNF alpha(-/-) and wild-type (WT) mice with implanted osmotic minipumps. Daily measurement of water intake, salt intake, and urine output were performed using metabolic cages. Blood pressure was monitored continuously with implanted radiotelemetry. Ang II administration for 2 weeks caused increases in salt (0.2+/ 0.07 to 5.6+/-0.95 mL/d) and water (5.4+/-0.34 to 11.5+/-1.2 mL/d) intake and in mean arterial pressure (115+/-1 to 151+/-3 mm Hg) in wild-type mice, but these responses were absent in TNF-alpha(-/-) mice (0.2+/-0.04 to 0.3+/-0.09 mL/d, 5.5+/-0.2 to 6.1+/-0.07 mL/d, and 113+/-2 to 123+/-3 mm Hg, respectively). Cardiac hypertrophy induced by Ang II was significantly attenuated in TNF-alpha( /-) mice compared with wild-type mice. In a group of TNF-alpha(-/-) mice, when replacement therapy was made with recombinant TNF-alpha, Ang II induced similar responses in salt appetite, mean arterial pressure, and cardiac hypertrophy, as observed in wild-type mice. These results suggest that TNF-alpha plays a mechanistic role in mediating chronic Ang II-induced effects on salt appetite and blood pressure, as well as on cardiac hypertrophy. PMID- 18391106 TI - Comparison of "risk-adjusted" hospital outcomes. AB - BACKGROUND: A frequent challenge in outcomes research is the comparison of rates from different populations. One common example with substantial health policy implications involves the determination and comparison of hospital outcomes. The concept of "risk-adjusted" outcomes is frequently misunderstood, particularly when it is used to justify the direct comparison of performance at 2 specific institutions. METHODS AND RESULTS: Data from 14 Massachusetts hospitals were analyzed for 4393 adults undergoing isolated coronary artery bypass graft surgery in 2003. Mortality estimates were adjusted using clinical data prospectively collected by hospital personnel and submitted to a data coordinating center designated by the state. The primary outcome was hospital-specific, risk standardized, 30-day all-cause mortality after surgery. Propensity scores were used to assess the comparability of case mix (covariate balance) for each Massachusetts hospital relative to the pool of patients undergoing coronary artery bypass grafting surgery at the remaining hospitals and for selected pairwise comparisons. Using hierarchical logistic regression, we indirectly standardized the mortality rate of each hospital using its expected rate. Predictive cross-validation was used to avoid underidentification of true outlying hospitals. Overall, there was sufficient overlap between the case mix of each hospital and that of all other Massachusetts hospitals to justify comparison of individual hospital performance with that of the remaining hospitals. As expected, some pairwise hospital comparisons indicated lack of comparability. This finding illustrates the fallacy of assuming that risk adjustment per se is sufficient to permit direct side-by-side comparison of healthcare providers. In some instances, such analyses may be facilitated by the use of propensity scores to improve covariate balance between institutions and to justify such comparisons. CONCLUSIONS: Risk-adjusted outcomes, commonly the focus of public report cards, have a specific interpretation. Using indirect standardization, these outcomes reflect a provider's performance for its specific case mix relative to the expected performance of an average provider for that same case mix. Unless study design or post hoc adjustments have resulted in reasonable overlap of case-mix distributions, such risk-adjusted outcomes should not be used to directly compare one institution with another. PMID- 18391107 TI - Neuronal nitric oxide synthase regulates basal microvascular tone in humans in vivo. AB - BACKGROUND: Nitric oxide (NO) has a pivotal role in the regulation of vascular tone and blood flow, with dysfunctional release contributing to disease pathophysiology. These effects have been attributed to NO production by the endothelial NO synthase (eNOS); however, recent evidence suggests that a neuronal NO synthase (nNOS) may also be expressed in arterial vessels. METHODS AND RESULTS: We undertook a first-in-humans investigation of the role of nNOS in the local regulation of vascular blood flow in healthy subjects. Brachial artery infusion of the nNOS-specific inhibitor S-methyl-L-thiocitrulline (SMTC, 0.025 micromol/min to 0.2 micromol/min) caused a dose-dependent reduction in basal flow, with a 30.1+/-3.8% decrease at the highest dose (n=10; mean+/-SE; P<0.01). The effect of SMTC was abolished by coinfusion of the NO synthase substrate L arginine but was unaffected by D-arginine. A similar reduction in basal flow with the nonselective NO synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA; 37.4+/ 3.1%, n=10) required a 20-fold higher dose of 4 micromol/min. At doses that produced comparable reductions in basal flow, only L-NMMA (4 micromol/min) and not SMTC (0.2 micromol/min) inhibited acetylcholine-induced vasodilation; however, both SMTC and L-NMMA inhibited the forearm vasodilator response to mental stress. CONCLUSIONS: Basal forearm blood flow in humans is regulated by nNOS-derived NO, in contrast to the acetylcholine-stimulated increase in blood flow, which, as shown previously, is mediated primarily by eNOS. These data indicate that vascular nNOS has a distinct local role in the physiological regulation of human microvascular tone in vivo. PMID- 18391108 TI - Death without prior appropriate implantable cardioverter-defibrillator therapy: a competing risk study. AB - BACKGROUND: Implantable cardioverter-defibrillators (ICDs) improve survival in selected patients with left ventricular systolic dysfunction in randomized trials. Competing death without prior appropriate ICD therapy might preclude benefit from ICD implantation in a less selected routine-care population. METHODS AND RESULTS: We selected all patients with ischemic or dilated cardiomyopathy with an ICD implanted for primary or secondary prevention from a single-center prospective registry between 1994 and 2006. The end point was time to first appropriate ICD therapy/confirmed ventricular fibrillation or death without prior appropriate ICD therapy. We analyzed cumulative incidence functions and used competing risk regression to study predictors of appropriate ICD therapy or prior death. In 442 patients, 73 deaths occurred during a median follow-up of 3.6 years (maximum, 12.7 years). The cumulative incidence of first appropriate ICD therapy until year 7 was 52%, whereas 11% died without prior ICD therapy. The cumulative incidence of appropriate ICD therapy for ventricular fibrillation was 13%, whereas 23% died without prior therapy for ventricular fibrillation. Appropriate ICD therapy was twice as likely in secondary prevention compared with primary prevention, whereas death rates before ICD therapy were similar in both groups. Diuretic use for heart failure compared with nonuse predicted a 4-fold-increased risk of death prior to ICD therapy, although the incidence of appropriate ICD therapy was similar in both groups. CONCLUSIONS: In a contemporary ICD population, the risk of death without prior appropriate ICD therapy is substantial, especially in patients with advanced heart failure. PMID- 18391109 TI - Percutaneous pulmonary valve implantation: impact of evolving technology and learning curve on clinical outcome. AB - BACKGROUND: Percutaneous pulmonary valve implantation was introduced in the year 2000 as a nonsurgical treatment for patients with right ventricular outflow tract dysfunction. METHODS AND RESULTS: Between September 2000 and February 2007, 155 patients with stenosis and/or regurgitation underwent percutaneous pulmonary valve implantation. This led to significant reduction in right ventricular systolic pressure (from 63+/-18 to 45+/-13 mm Hg, P<0.001) and right ventricular outflow tract gradient (from 37+/-20 to 17+/-10 mm Hg, P<0.001). Follow-up ranged from 0 to 83.7 months (median 28.4 months). Freedom from reoperation was 93% (+/ 2%), 86% (+/-3%), 84% (+/-4%), and 70% (+/-13%) at 10, 30, 50, and 70 months, respectively. Freedom from transcatheter reintervention was 95% (+/-2%), 87% (+/ 3%), 73% (+/-6%), and 73% (+/-6%) at 10, 30, 50, and 70 months, respectively. Survival at 83 months was 96.9%. On time-dependent analysis, the first series of 50 patients (log-rank test P<0.001) and patients with a residual gradient >25 mm Hg (log-rank test P=0.01) were associated with a higher risk of reoperations. CONCLUSIONS: Percutaneous pulmonary valve implantation resulted in the ability to avoid surgical right ventricular outflow tract revision in the majority of cases. This procedure might reduce the number of operations needed over the total lifetime of patients with right ventricle-to-pulmonary artery conduits. PMID- 18391110 TI - Enzymatic activity of lysosomal carboxypeptidase (cathepsin) A is required for proper elastic fiber formation and inactivation of endothelin-1. AB - BACKGROUND: Lysosomal carboxypeptidase, cathepsin A (protective protein, CathA), is a component of the lysosomal multienzyme complex along with beta-galactosidase (GAL) and sialidase Neu1, where it activates Neu1 and protects GAL and Neu1 against the rapid proteolytic degradation. On the cell surface, CathA, Neu1, and the enzymatically inactive splice variant of GAL form the elastin-binding protein complex. In humans, genetic defects of CathA cause galactosialidosis, a metabolic disease characterized by combined deficiency of CathA, GAL, and Neu1 and a lysosomal storage of sialylated glycoconjugates. However, several phenotypic features of galactosialidosis patients, including hypertension and cardiomyopathies, cannot be explained by the lysosomal storage. These observations suggest that CathA may be involved in hemodynamic functions that go beyond its protective activity in the lysosome. METHODS AND RESULTS: We generated a gene-targeted mouse in which the active CathA was replaced with a mutant enzyme carrying a Ser190Ala substitution in the active site. These animals expressed physiological amounts of catalytically inactive CathA protein, capable of forming lysosomal multienzyme complex, and did not develop secondary deficiency of Neu1 and GAL. Conversely, the mice showed a reduced degradation rate of the vasoconstrictor peptide, endothelin-1, and significantly increased arterial blood pressure. CathA-deficient mice also displayed scarcity of elastic fibers in lungs, aortic adventitia, and skin. CONCLUSIONS: Our results provide the first evidence that CathA acts in vivo as an endothelin-1-inactivating enzyme and strongly confirm a crucial role of this enzyme in effective elastic fiber formation. PMID- 18391111 TI - Inhaled nitric oxide enables artificial blood transfusion without hypertension. AB - BACKGROUND: One of the major obstacles hindering the clinical development of a cell-free, hemoglobin-based oxygen carrier (HBOC) is systemic vasoconstriction. METHODS AND RESULTS: Experiments were performed in healthy mice and lambs by infusion of either murine tetrameric hemoglobin (0.48 g/kg) or glutaraldehyde polymerized bovine hemoglobin (HBOC-201, 1.44 g/kg). We observed that intravenous infusion of either murine tetrameric hemoglobin or HBOC-201 induced prolonged systemic vasoconstriction in wild-type mice but not in mice congenitally deficient in endothelial nitric oxide (NO) synthase (NOS3). Treatment of wild type mice by breathing NO at 80 ppm in air for 15 or 60 minutes or with 200 ppm NO for 7 minutes prevented the systemic hypertension induced by subsequent intravenous administration of murine tetrameric hemoglobin or HBOC-201 and did not result in conversion of plasma hemoglobin to methemoglobin. Intravenous administration of sodium nitrite (48 nmol) 5 minutes before infusion of murine tetrameric hemoglobin also prevented the development of systemic hypertension. In awake lambs, breathing NO at 80 ppm for 1 hour prevented the systemic hypertension caused by subsequent infusion of HBOC-201. CONCLUSIONS: These findings demonstrate that HBOC can cause systemic vasoconstriction by scavenging NO produced by NOS3. Moreover, in 2 species, inhaled NO administered before the intravenous infusion of HBOC can prevent systemic vasoconstriction without causing methemoglobinemia. PMID- 18391112 TI - Comparison of coronary artery stenting outcomes in the eras before and after the introduction of drug-eluting stents. AB - BACKGROUND: Few studies have compared medium-term outcomes for drug-eluting stents (DES) and bare metal stents, and most are relatively small randomized controlled trials. Furthermore, since the introduction of DES, there has been increased use and duration of use of clopidogrel, statins, and other evidence based therapies. The purpose of the present study was to compare outcomes for patients who underwent stenting in the eras before and after the introduction of DES. METHODS AND RESULTS: New York state patients undergoing stenting in all nonfederal hospitals in the state were studied. Patients were excluded if they had a previous revascularization. Risk factors that were significant predictors of adverse outcomes were used to adjust adverse outcome rates. The study included 11,436 patients who received stents between October 1, 2002, and March 31, 2003, and 12,926 patients who underwent stenting between October 1, 2003, and March 31, 2004. Death rates, the combined end point of death and myocardial infarction (MI), nonfatal MI requiring readmission, target vessel revascularization, and target lesion revascularization were compared at 2 years. Patients in the DES era had significantly better risk-adjusted outcomes for death/MI (adjusted hazard ratio, 0.90; 95% confidence interval, 0.83 to 0.97), 9.9% versus 10.8%; nonfatal MI requiring readmission (adjusted hazard ratio, 0.86; 95% confidence interval, 0.76 to 0.97); target vessel revascularization (adjusted hazard ratio, 0.60; 95% confidence interval, 0.56 to 0.64), 11.2% versus 17.9%; and target lesion revascularization (hazard ratio, 0.55; 95% confidence interval, 0.51 to 0.59), 8.4% versus 14.7%. CONCLUSIONS: Patients in the DES era experienced lower rates of death/MI, nonfatal MI, target vessel revascularization, and target lesion revascularization, but there were no differences in the rates of death. These improvements are likely a result of increased use of clopidogrel, statins, and dual antiplatelet therapy, in addition to the introduction of DES. PMID- 18391113 TI - Absence of cyclooxygenase-2 exacerbates hypoxia-induced pulmonary hypertension and enhances contractility of vascular smooth muscle cells. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) is upregulated in pulmonary artery smooth muscle cells (PASMCs) during hypoxia and may play a protective role in the response of the lung to hypoxia. Selective COX-2 inhibition may have detrimental pulmonary vascular consequences during hypoxia. METHODS AND RESULTS: To investigate the role of COX-2 in the pulmonary vascular response to hypoxia, we subjected wild-type and COX-2-deficient mice to a model of chronic normobaric hypoxia. COX-2-null mice developed severe pulmonary hypertension with exaggerated elevation of right ventricular systolic pressure, significant right ventricular hypertrophy, and striking vascular remodeling after hypoxia. Pulmonary vascular remodeling in COX-2-deficient mice was characterized by PASMC hypertrophy but not increased proliferation. Furthermore, COX-2-deficient mice had significant upregulation of the endothelin-1 receptor (ET(A)) in the lung after hypoxia. Similarly, selective pharmacological inhibition of COX-2 in wild-type mice exacerbated hypoxia-induced pulmonary hypertension and resulted in PASMC hypertrophy and increased ET(A) receptor expression in pulmonary arterioles. The absence of COX-2 in vascular smooth muscle cells during hypoxia in vitro augmented traction forces and enhanced contractility of an extracellular matrix. Treatment of COX-2-deficient PASMCs with iloprost, a prostaglandin I(2) analog, and prostaglandin E(2) abrogated the potent contractile response to hypoxia and restored the wild-type phenotype. CONCLUSIONS: Our findings reveal that hypoxia induced pulmonary hypertension and vascular remodeling are exacerbated in the absence of COX-2 with enhanced ET(A) receptor expression and increased PASMC hypertrophy. COX-2-deficient PASMCs have a maladaptive response to hypoxia manifested by exaggerated contractility, which may be rescued by either COX-2 derived prostaglandin I(2) or prostaglandin E(2). PMID- 18391114 TI - Prevention of heart failure: a scientific statement from the American Heart Association Councils on Epidemiology and Prevention, Clinical Cardiology, Cardiovascular Nursing, and High Blood Pressure Research; Quality of Care and Outcomes Research Interdisciplinary Working Group; and Functional Genomics and Translational Biology Interdisciplinary Working Group. AB - The increase in heart failure (HF) rates throughout the developed and developing regions of the world poses enormous challenges for caregivers, researchers, and policymakers. Therefore, prevention of this global scourge deserves high priority. Identifying and preventing the well-recognized illnesses that lead to HF, including hypertension and coronary heart disease, should be paramount among the approaches to prevent HF. Aggressive implementation of evidence-based management of risk factors for coronary heart disease should be at the core of HF prevention strategies. Questions currently in need of attention include how to identify and treat patients with asymptomatic left ventricular systolic dysfunction (Stage B HF) and how to prevent its development. The relationship of chronic kidney disease to HF and control of chronic kidney disease in prevention of HF need further investigation. Currently, we have limited understanding of the pathophysiological basis of HF in patients with preserved left ventricular systolic function and management techniques to prevent it. New developments in the field of biomarker identification have opened possibilities for the early detection of individuals at risk for developing HF (Stage A HF). Patient groups meriting special interest include the elderly, women, and ethnic/racial minorities. Future research ought to focus on obtaining a much better knowledge of genetics and HF, especially both genetic risk factors for development of HF and genetic markers as tools to guide prevention. Lastly, a national awareness campaign should be created and implemented to increase public awareness of HF and the importance of its prevention. Heightened public awareness will provide a platform for advocacy to create national research programs and healthcare policies dedicated to the prevention of HF. PMID- 18391115 TI - Noninvasive detection and localization of vulnerable plaque and arterial thrombosis with computed tomography angiography/positron emission tomography. AB - BACKGROUND: It has been shown that plaque uptake of fluorodeoxyglucose is proportional to macrophage density. We tested the hypothesis that arterial thrombosis occurs in areas with high fluorodeoxyglucose uptake and that computed tomography angiography (CTA) can detect thrombi in vessels. METHODS AND RESULTS: Twenty New Zealand White rabbits were studied before and after atherosclerosis induction through de-endothelialization and a high-cholesterol diet; 14 were then thrombus triggered. CTA/positron emission tomography scans were performed before cholesterol diet, at the middle diet feeding, at the end of diet feeding, and after triggering. Serum inflammatory markers were measured. Maximal standardized uptake value was measured over the thoracic and upper and lower abdominal aortas and correlated with thrombosis and macrophage density on sections from the same sites. Aortic diameters averaged 2.84+/-1.16 mm. The sensitivity, specificity, and accuracy of CTA for detecting thrombi were 92%, 89%, and 90%, respectively. Plasminogen activator inhibitor-1 and C-reactive protein levels increased with atherosclerosis and thrombosis triggering. Maximal standardized uptake value at baseline was 0.62+/-0.13, 0.96+/-0.33 at the middle of feeding, and 1.06+/-0.38 at the end of feeding. Segments that developed thrombosis had the highest maximal standardized uptake value of 1.32+/-0.69 (113% increase; P=0.002) and had a 129% increase in macrophage density compared with segments without thrombi (P=0.01). CONCLUSIONS: Fluorodeoxyglucose uptake was proportional to the duration of cholesterol feeding and peaked with plaque disruption and thrombosis. CTA was highly accurate in detecting thrombi. Our findings in this animal model of atherosclerotic plaques with high macrophage density showed that CTA/positron emission tomography can be used to identify and localize inflamed plaques and thrombosis. With the currently available technology and nuclear tracers, however, many challenges remain before clinical applications are possible. PMID- 18391116 TI - Images in cardiovascular medicine. Real-time 3-dimensional transesophageal echocardiography: novel utility in atrial fibrillation ablation with a prosthetic mitral valve. PMID- 18391117 TI - Letter by Sadek and Garry regarding article, "Iron-oxide labeling and outcome of transplanted mesenchymal stem cells in the infarcted myocardium". PMID- 18391119 TI - How does folic acid cure heart attacks? PMID- 18391120 TI - Disease-modifying mutations in familial hypertrophic cardiomyopathy: complexity from simplicity. PMID- 18391121 TI - Modified prosthetic vascular conduits. AB - Atherosclerosis in the form of peripheral arterial disease results in significant morbidity. Surgical treatment options for peripheral arterial disease include angioplasty, endarterectomy, and bypass grafting. For bypass grafting, vein remains the conduit of choice; however, poor quality and limited availability have led to the use of prosthetic materials. Unfortunately, because of a lack of endothelium and compliance mismatch, neointimal hyperplasia develops aggressively, resulting in high failure rates. To improve graft patency, investigators have developed surgical, chemical, and biological graft modifications. This review describes common prosthetic materials, as well as approaches currently in use and under investigation to modify and improve prosthetic conduits for bypass grafting in an effort to improve graft patency rates. PMID- 18391123 TI - Induced Brugada-type electrocardiogram, a sign for imminent malignant arrhythmias. PMID- 18391124 TI - Images in cardiovascular medicine. Fatal pulmonary lymphangiectasia manifesting after repeated surgeries for intractable chylopericardium and chylothorax in a 20 month-old girl. PMID- 18391122 TI - Medical management of small abdominal aortic aneurysms. AB - Abdominal aortic aneurysm is a common condition that may be lethal when it is unrecognized. Current guidelines suggest repair as the aneurysm diameter reaches 5.0 to 5.5 cm. Most aortic aneurysms are detected incidentally when imaging is done for other purposes or through screening programs. Ninety percent of these aneurysms are below the threshold for intervention at the time of detection. A number of studies have sought to determine factors that lead to progression of aneurysmal disease that might be amenable to intervention during this period of observation. We review these studies and make recommendations for the medical management of small abdominal aortic aneurysms. On the basis of our current knowledge of the causes of aneurysm, a number of approaches have been proposed to prevent progression of aneurysmal disease. These include hemodynamic management, inhibition of inflammation, and protease inhibition. The American College of Cardiology/American Heart Association clinical practice guidelines rules of evidence have helped to define strength of evidence to support these approaches. Level A evidence (from large randomized trials) is available to indicate that observation of small aneurysms in men is safe up to a size of 5.5 cm and that propranolol does not inhibit aneurysm expansion. Level B evidence (from small randomized trials) suggests that roxithromycin or doxycycline will decrease the rate of aneurysm expansion. A number of studies agree that tobacco use is associated with an increased rate of aneurysm expansion. Level B and C evidence is available to suggest that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) may inhibit aneurysm expansion. There are animal data but no human data demonstrating that angiotensin-converting enzyme inhibitors or losartan, an angiotensin receptor blocker, will decrease the rate of AAA expansion. A pharmacological agent without important side effects that inhibited aneurysm expansion could change current approaches to aneurysm treatment. Additional studies are needed to clarify the potential role of doxycycline, roxithromycin, and statin therapy in the progression of aneurysmal disease. PMID- 18391125 TI - The suicide of Lucretia. PMID- 18391126 TI - Dysfunctional neural plasticity in patients with schizophrenia. AB - CONTEXT: Neural plasticity in the human cortex involves a reorganization of synaptic connections in an effort to adapt to a changing environment. In schizophrenia, dysfunctional neural plasticity has been proposed as a key pathophysiological mechanism. OBJECTIVE: To evaluate neural plasticity in unmedicated and medicated patients with schizophrenia compared with healthy subjects. DESIGN: Neural plasticity can be evaluated from the motor cortex in healthy subjects using transcranial magnetic stimulation through a paradigm known as use-dependent plasticity. This paradigm involves several steps: (1) measuring the spontaneous direction of transcranial magnetic stimulation-induced thumb movements; (2) training subjects to practice thumb movements opposite to this baseline direction for 30 minutes; and (3) measuring the direction of transcranial magnetic stimulation-induced thumb movement after training. Previous experiments have shown that in healthy subjects, posttraining transcranial magnetic stimulation-induced movements occur in a vector commensurate with the practiced movements, which may be associated with time-limited reorganization of motor circuits. SETTING: All of the participants were recruited and evaluated at the Centre for Addiction and Mental Health. PARTICIPANTS: Fourteen medicated and 6 unmedicated patients with schizophrenia and 20 healthy subjects were recruited. MAIN OUTCOME MEASURE: It was anticipated that patients with schizophrenia would demonstrate attenuated motor reorganization in the direction of training. RESULTS: Both medicated and unmedicated patients with schizophrenia demonstrated significantly reduced motor reorganization compared with healthy subjects. CONCLUSIONS: It is possible that in schizophrenia, these deficits in neural plasticity are related to disturbances of gamma-aminobutyric acid, N-methyl-D aspartate neurotransmission, or dopamine that may potentially account for the aberrant motor performance of these patients. PMID- 18391127 TI - Residual symptom recovery from major affective episodes in bipolar disorders and rapid episode relapse/recurrence. AB - CONTEXT: Both bipolar disorder type I and type II are characterized by frequent affective episode relapse and/or recurrence. An increasingly important goal of therapy is reducing chronicity by preventing or delaying additional episodes. OBJECTIVES: To determine whether the continued presence of subsyndromal residual symptoms during recovery from major affective episodes in bipolar disorder is associated with significantly faster episode recurrence than asymptomatic recovery and whether this is the strongest correlate of early episode recurrence among 13 variables examined. DESIGN: An ongoing prospective, naturalistic, and systematic 20-year follow-up investigation of mood disorders: the National Institute of Mental Health Collaborative Depression Study. SETTING: Five academic tertiary care centers. PARTICIPANTS: Two hundred twenty-three participants with bipolar disorder (type I or II) were followed up prospectively for a median of 17 years (mean, 14.1 [SD, 6.2] years). MAIN OUTCOME MEASURE: Participants defined as recovered by Research Diagnostic Criteria from their index major depressive episode and/or mania were divided into residual vs asymptomatic recovery groups and were compared according to the time to their next major affective episodes. RESULTS: Participants recovering with residual affective symptoms experienced subsequent major affective episodes more than 3 times faster than asymptomatic recoverers (hazard ratio, 3.36; 95% confidence interval, 2.25-4.98; P < .001). Recovery status was the strongest correlate of time to episode recurrence (P < .001), followed by a history of 3 or more affective episodes before intake (P = .007). No other variable examined was significantly associated with time to recurrence. CONCLUSIONS: In bipolar disorder, residual symptoms after resolution of a major affective episode indicate that the individual is at significant risk for a rapid relapse and/or recurrence, suggesting that the illness is still active. Stable recovery in bipolar disorder is achieved only when asymptomatic status is achieved. PMID- 18391128 TI - A discriminating messenger RNA signature for bipolar disorder formed by an aberrant expression of inflammatory genes in monocytes. AB - CONTEXT: Mood disturbances are associated with an activated inflammatory response system. OBJECTIVE: To identify a discriminating and coherent expression pattern of proinflammatory genes in monocytes of patients with bipolar disorder. DESIGN: A quantitative polymerase chain reaction (Q-PCR) case-control gene expression study on purified monocytes of bipolar patients, the offspring of bipolar patients, and healthy control participants after having selected 22 discriminating inflammatory genes using whole genome analyses. SETTING: Academic research setting in The Netherlands. PATIENTS: Forty-two bipolar patients with 25 healthy controls, 54 offspring of a bipolar parent (13 had a mood disorder and 3 developed one during follow-up), and 70 healthy children underwent Q-PCR. MAIN OUTCOME MEASURE: Inflammatory gene expression levels in monocytes. RESULTS: We detected in the monocytes of bipolar patients a coherent mutually correlating set (signature) of 19 aberrantly expressed (P < .01) messenger RNAs of inflammatory (PDE4B, IL1B, IL6, TNF, TNFAIP3, PTGS2, and PTX3), trafficking (CCL2, CCL7, CCL20, CXCL2, CCR2, and CDC42), survival (BCL2A1 and EMP1), and mitogen-activated protein kinase pathway (MAPK6, DUSP2, NAB2, and ATF3) genes. Twenty-three of 42 bipolar patients (55%) had a positive signature test result vs 7 of 38 healthy controls (18%) (positive test result: positivity for PDE4B, ie, a messenger RNA 1 SD higher than the mean level found in healthy controls, plus 25% of the other genes with similar positive findings). Positive signature test results were also present in 11 of 13 offspring with a mood disorder (85%), 3 of 3 offspring developing a mood disorder (100%), and 17 of 38 euthymic offspring (45%) vs 13 of 70 healthy children (19%). Lithium carbonate and antipsychotic treatment downregulated the gene expression of most inflammatory genes. CONCLUSIONS: The monocytes of a large proportion of bipolar patients and offspring of bipolar parents showed an inflammatory gene expression signature. This coherent set of genes opens new avenues for biomarker development with possibilities for disease prediction in individuals genetically at risk and for the subclassification of bipolar patients who could possibly benefit from anti-inflammatory treatment. PMID- 18391129 TI - Elevated inflammation levels in depressed adults with a history of childhood maltreatment. AB - CONTEXT: The association between depression and inflammation is inconsistent across research samples. OBJECTIVE: To test whether a history of childhood maltreatment could identify a subgroup of depressed individuals with elevated inflammation levels, thus helping to explain previous inconsistencies. DESIGN: Prospective longitudinal cohort study. SETTING: New Zealand. PARTICIPANTS: A representative birth cohort of 1000 individuals was followed up to age 32 years as part of the Dunedin Multidisciplinary Health and Development Study. Study members were assessed for history of childhood maltreatment and current depression. MAIN OUTCOME MEASURES: Inflammation was assessed using a clinically relevant categorical measure of high-sensitivity C-reactive protein (>3 mg/L) and a dimensional inflammation factor indexing the shared variance of continuous measures of high-sensitivity C-reactive protein, fibrinogen, and white blood cells. RESULTS: Although depression was associated with high levels of high sensitivity C-reactive protein (relative risk,1.45; 95% confidence interval,1.06 1.99), this association was significantly attenuated and no longer significant when the effect of childhood maltreatment was taken into account. Individuals with current depression and a history of childhood maltreatment were more likely to have high levels of high-sensitivity C-reactive protein compared with control subjects (n = 27; relative risk, 2.07; 95% confidence interval, 1.23-3.47). In contrast, individuals with current depression only had a nonsignificant elevation in risk (n = 109; relative risk, 1.40; 95% confidence interval, 0.97-2.01). Results were generalizable to the inflammation factor. The elevated inflammation levels in individuals who were both depressed and maltreated were not explained by correlated risk factors such as depression recurrence, low socioeconomic status in childhood or adulthood, poor health, or smoking. CONCLUSIONS: A history of childhood maltreatment contributes to the co-occurrence of depression and inflammation. Information about experiences of childhood maltreatment may help to identify depressed individuals with elevated inflammation levels and, thus, at greater risk of cardiovascular disease. PMID- 18391130 TI - A longitudinal twin study of fears from middle childhood to early adulthood: evidence for a developmentally dynamic genome. AB - CONTEXT: While the nature of common fears changes over development, we do not know whether genetic effects on fear-proneness are developmentally stable or developmentally dynamic. OBJECTIVE: To determine the temporal pattern of genetic and environmental effects on the level of intensity of common fears. DESIGN: Prospective, 4-wave longitudinal twin study. Structural modeling was performed with Mx. SETTING: General community. PARTICIPANTS: Two thousand four hundred ninety twins and their parents from the Swedish Twin Study of Child and Adolescent Development. MAIN OUTCOME MEASURE: The level of parent- and/or self reported fears obtained at ages 8 to 9, 13 to 14, 16 to 17, and 19 to 20 years. RESULTS: Thirteen questionnaire items formed 3 distinct fear factors: situational, animal, and blood/injury. For all 3 fears, the best-fit model revealed developmentally dynamic effects and, in particular, evidence for both genetic attenuation and innovation. That is, genetic factors influencing fear intensity at age 8 to 9 years decline substantially in importance over time. Furthermore, new sets of genetic risk factors impacting fear intensity "come on line" in early adolescence, late adolescence, and early adulthood. As the twins aged, the influence of the shared environment declined and unique environment increased. No sex effects were found for situational fears while for animal and blood/injury fears, genetic factors in males and females were correlated but not identical. Shared environmental factors were both more important and more stable for animal fears than for situational or blood/injury fears. CONCLUSIONS: Genetic effects on fear are developmentally dynamic from middle childhood to young adulthood. As children age, familial-environmental influences on fears decline in importance. PMID- 18391131 TI - A second look at prior trauma and the posttraumatic stress disorder effects of subsequent trauma: a prospective epidemiological study. AB - CONTEXT: Previous studies showed increased probability of a posttraumatic stress disorder (PTSD) effect of trauma in persons who had experienced prior trauma. The evidence comes chiefly from retrospective data on earlier events, obtained from trauma-exposed persons with and without PTSD. A generally overlooked major limitation is the failure to assess the PTSD response to the prior trauma. OBJECTIVE: To estimate the risk of PTSD after traumas experienced during follow up periods in relation to respondents' prior traumatic events and PTSD. DESIGN: A cohort study of young adults interviewed initially in 1989, with repeated assessments during a 10-year follow-up. SETTING AND PARTICIPANTS: The sample was randomly selected from a large health maintenance organization in Southeast Michigan, representing the geographic area. MAIN OUTCOME MEASURES: The relative risk of PTSD precipitated by traumatic events occurring during follow-up periods in relation to prior exposure and PTSD that had occurred during preceding periods, estimated by general estimating equations (n = 990). RESULTS: The conditional risk of PTSD during the follow-up periods was significantly higher among trauma-exposed persons who had experienced prior PTSD, relative to those with no prior trauma (odds ratio, 3.01; 95% confidence interval, 1.52-5.97). After adjustment for sex, race, education, and preexisting major depression and anxiety disorders, the estimates were only marginally revised. In contrast, the conditional risk of PTSD during follow-up among trauma-exposed persons who had experienced prior traumatic events but not PTSD was not significantly elevated, relative to trauma-exposed persons with no prior trauma. The difference between the 2 estimates was significant (P = .005). CONCLUSIONS: Prior trauma increases the risk of PTSD after a subsequent trauma only among persons who developed PTSD in response to the prior trauma. The findings suggest that preexisting susceptibility to a pathological response to stressors may account for the PTSD response to the prior trauma and the subsequent trauma. PMID- 18391132 TI - Change in depressive symptoms during the prodromal phase of Alzheimer disease. AB - CONTEXT: Prospective studies have established an association between depressive symptoms and risk of dementia, but how depressive symptoms change during the evolution of dementia is uncertain. OBJECTIVE: To test the hypothesis that depressive symptoms increase during the prodromal phase of Alzheimer disease (AD). DESIGN: Prospective cohort study. PARTICIPANTS AND SETTING: For up to 13 years, 917 older Catholic nuns, priests, and monks without dementia at study onset completed annual clinical evaluations that included administration of the 10-item Center for Epidemiologic Studies Depression Scale and clinical classification of mild cognitive impairment and AD. MAIN OUTCOME MEASURE: Change in depressive symptoms reported on the Center for Epidemiologic Studies Depression Scale. RESULTS: At baseline, participants reported a mean (SD) of 1.0 (1.5) depressive symptoms. Those who developed AD (n = 190) showed no increase in depressive symptoms before the diagnosis was made, and this finding was not modified by age, sex, education, memory complaints, vascular burden, or personality. There was no systematic change in depressive symptoms after the AD diagnosis, although symptoms tended to decrease in women relative to men and in those with a higher premorbid level of openness and a lower premorbid level of agreeableness. Among those without cognitive impairment at baseline, depressive symptoms did not increase in those who subsequently developed mild cognitive impairment. CONCLUSION: We found no evidence of an increase in depressive symptoms during the prodromal phase of AD. PMID- 18391133 TI - Achievement and maintenance of sustained response during the Treatment for Adolescents With Depression Study continuation and maintenance therapy. AB - CONTEXT: The Treatment for Adolescents With Depression Study evaluated fluoxetine (FLX), cognitive behavioral therapy (CBT), and FLX/CBT combination (COMB) vs pill placebo in 439 adolescents with major depressive disorder. Treatment consisted of 3 stages: (1) acute (12 weeks), (2) continuation (6 weeks), and (3) maintenance (18 weeks). OBJECTIVE: To examine rates of achieving and maintaining sustained response during continuation and maintenance treatments. DESIGN: Randomized controlled trial. Response was determined by blinded independent evaluators. SETTING: Thirteen US sites. PATIENTS: Two hundred forty-two FLX, CBT, and COMB patients in their assigned treatment at the end of stage 1. INTERVENTIONS: Stage 2 treatment varied based on stage 1 response. Stage 3 consisted of 3 CBT and/or pharmacotherapy sessions and, if applicable, continued medication. MAIN OUTCOME MEASURES: Sustained response was defined as 2 consecutive Clinical Global Impression-Improvement ratings of 1 or 2 ("full response"). Patients achieving sustained response were classified on subsequent nonresponse status. RESULTS: Among 95 patients (39.3%) who had not achieved sustained response by week 12 (29.1% COMB, 32.5% FLX, and 57.9% CBT), sustained response rates during stages 2 and 3 were 80.0% COMB, 61.5% FLX, and 77.3% CBT (difference not significant). Among the remaining 147 patients (60.7%) who achieved sustained response by week 12, CBT patients were more likely than FLX patients to maintain sustained response through week 36 (96.9% vs 74.1%; P = .007; 88.5% of COMB patients maintained sustained response through week 36). Total rates of sustained response by week 36 were 88.4% COMB, 82.5% FLX, and 75.0% CBT. CONCLUSIONS: Most adolescents with depression who had not achieved sustained response during acute treatment did achieve that level of improvement during continuation and maintenance therapies. The possibility that CBT may help the subset of adolescents with depression who achieve early sustained response maintain their response warrants further investigation. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00006286. PMID- 18391134 TI - Comparing drug-related hospital morbidity following heroin dependence treatment with methadone maintenance or naltrexone implantation. AB - CONTEXT: Most research on heroin dependence treatments assesses short-term changes in patients' self-reported drug use. To our knowledge, long-term sustainability of changes in patients' drug use and associated hospital morbidity posttreatment have not been studied. OBJECTIVES: To evaluate drug-related hospital morbidity in heroin users at 6 months and 3 1/2 years after receiving naltrexone implant treatment and to compare these results with outcomes from a similar cohort treated with methadone maintenance treatment. DESIGN: Retrospective longitudinal follow-up, using data prospectively collected via a state hospital (public and private) reporting system. SETTING: Perth, Western Australia. Methadone maintenance dosage was generally dispensed daily by registered community pharmacies. Naltrexone implant treatment was performed as a day procedure at a community clinic. PARTICIPANTS: A total of 522 and 314 heroin dependent persons (according to DSM-IV), first time treated with methadone maintenance or a naltrexone implant, respectively, between January 1, 2001, and December 30, 2002, were identified, using health record linkage. MAIN OUTCOME MEASURES: Planned outcomes included crude hospital admission rates, adjusted changes in risks (odds ratio [OR]), and rates (rate ratio) of "overdose-related" and "non-overdose-related" hospital morbidity associated with opioid vs nonopioid drugs 6 months and 3 1/2 years posttreatment. RESULTS: Following naltrexone implant treatment, opioid-related risk decreased for overdose (OR, 0.23; 95% confidence interval [CI], 0.11-0.48) and nonoverdose (OR, 0.64; 95% CI, 0.46 0.89) conditions at 3 1/2 years. Such reductions were not observed after methadone treatment. Overdose on nonopioid drugs increased in older patients to 6 months: OR of 16.31 (95% CI, 3.07-86.53) for naltrexone and OR of 5.03 (95% CI, 1.18-21.54) for methadone. Nonoverdose (eg, dependence and withdrawal) associated with nonopioid drugs also increased for patients receiving the naltrexone implant: OR of 1.52 (95% CI, 1.04-2.23) at 3 1/2 years. In addition, there were 6 drug-related deaths: 5 after methadone maintenance and 1 after naltrexone implantation. CONCLUSIONS: Naltrexone implants, but not methadone maintenance, has long-term benefits in reducing opioid-related hospital morbidity. However, long-lasting and increased nonopioid drug-related morbidity following naltrexone implantation is particularly concerning. Similar studies are required to confirm these findings. PMID- 18391136 TI - Rapid increase in rates of bipolar diagnosis in youth: "true" bipolarity or misdiagnosed severe disruptive behavior disorders? PMID- 18391135 TI - Effect of naltrexone and ondansetron on alcohol cue-induced activation of the ventral striatum in alcohol-dependent people. AB - CONTEXT: Medication for the treatment of alcoholism is currently not particularly robust. Neuroimaging techniques might predict which medications could be useful in the treatment of alcohol dependence. OBJECTIVE: To explore the effect of naltrexone, ondansetron hydrochloride, or the combination of these medications on cue-induced craving and ventral striatum activation. DESIGN: Functional brain imaging was conducted during alcohol cue presentation. SETTING: Participants were recruited from the general community following media advertisement. Experimental procedures were performed in the magnetic resonance imaging suite of a major training hospital and medical research institute. PATIENTS: Ninety non-treatment seeking alcohol-dependent (by DSM-IV criteria) and 17 social drinking (< 14 drinks per week) paid volunteers recruited through advertisements at an academic center. INTERVENTIONS: A taste of alcohol and a series of alcohol-related pictures, neutral beverage pictures, and visual control images were provided to volunteers after 7 days of double-blind randomly assigned daily dosing with 50 mg of naltrexone (n = 23), 0.50 mg of ondansetron hydrochloride (n = 23), the combination of the 2 medications (n = 20), or matching placebos (n = 24). MAIN OUTCOME MEASURES: Difference in brain blood oxygen level-dependent magnetic resonance when viewing alcohol pictures vs neutral beverage pictures with a particular focus on ventral striatum activity comparison across medication groups. Self-ratings of alcohol craving. RESULTS: The combination treatment decreased craving for alcohol. Naltrexone with (P = .02) or without (P = .049) ondansetron decreased alcohol cue-induced activation of the ventral striatum. Ondansetron by itself was similar to naltrexone and the combination in the overall analysis but intermediate in a region-specific analysis. CONCLUSIONS: Consistent with animal data that suggest that both naltrexone and ondansetron reduce alcohol-stimulated dopamine output in the ventral striatum, the current study found evidence that these medications, alone or in combination, could decrease alcohol cue-induced activation of the ventral striatum, consistent with their putative treatment efficacy. PMID- 18391137 TI - A genetic decomposition of the association between parasomnias and dyssomnias in 8-year-old twins. AB - OBJECTIVE: To estimate genetic and environmental influences on parasomnias and dyssomnias and their association in 8-year-olds. DESIGN: Parents of twins completed the Children's Sleep Habits Questionnaire. SETTING: Families were primarily tested at the Institute of Psychiatry, London, England. PARTICIPANTS: A total of 300 pairs of 8-year-old twins and their parents participated in the study. MAIN OUTCOME MEASURES: Sleep difficulties in children. RESULTS: Individual differences in parasomnias and dyssomnias were largely explained by genes (accounting for 50% and 71% of the variances, respectively). The rest of the variances were mainly due to nonshared environmental influences. A moderate association was found between parasomnias and dyssomnias (r = 0.42), which was mainly accounted for by genetic influences (87%). The genetic correlation between parasomnias and dyssomnias was moderate (r = 0.61). In contrast, the nonshared environmental correlation was small (r = 0.10). CONCLUSIONS: The decomposition of the association between parasomnias and dyssomnias suggests that there may be different manifestations of shared underlying genetic risks for sleep problems partly dependent on nonshared environmental influence. PMID- 18391138 TI - Short sleep duration in infancy and risk of childhood overweight. AB - OBJECTIVE: To examine the extent to which infant sleep duration is associated with overweight at age 3 years. DESIGN: Longitudinal survey. SETTING: Multisite group practice in Massachusetts. PARTICIPANTS: Nine hundred fifteen children in Project Viva, a prospective cohort. Main Exposure At children's ages 6 months, 1 year, and 2 years, mothers reported the number of hours their children slept in a 24-hour period, from which we calculated a weighted average of daily sleep. MAIN OUTCOME MEASURES: We used multivariate regression analyses to predict the independent effects of sleep duration (< 12 h/d vs > or = 12 h/d) on body mass index (BMI) (calculated as the weight in kilograms divided by the height in meters squared) z score, the sum of subscapular and triceps skinfold thicknesses, and overweight (BMI for age and sex > or = 95th percentile) at age 3 years. RESULTS: The children's mean (SD) duration of daily sleep was 12.3 (1.1) hours. At age 3 years, 83 children (9%) were overweight; the mean (SD) BMI z score and sum of subscapular and triceps skinfold thicknesses were 0.44 (1.03) and 16.66 (4.06) mm, respectively. After adjusting for maternal education, income, prepregnancy BMI, marital status, smoking history, and breastfeeding duration and child's race/ethnicity, birth weight, 6-month weight-for-length z score, daily television viewing, and daily participation in active play, we found that infant sleep of less than 12 h/d was associated with a higher BMI z score (beta, 0.16; 95% confidence interval, 0.02-0.29), higher sum of subscapular and triceps skinfold thicknesses (beta, 0.79 mm; 95% confidence interval, 0.18-1.40), and increased odds of overweight (odds ratio, 2.04; 95% confidence interval, 1.07 3.91). CONCLUSION: Daily sleep duration of less than 12 hours during infancy appears to be a risk factor for overweight and adiposity in preschool-aged children. PMID- 18391139 TI - Effect of weight, sleep duration, and comorbid sleep disorders on behavioral outcomes in children with sleep-disordered breathing. AB - OBJECTIVE: To assess the relative contribution of potential risk factors for adverse neurobehavioral outcomes in children referred for evaluation of sleep disordered breathing (SDB), including weight, mean sleep duration, and comorbid sleep disorders. DESIGN: Medical record review. SETTING: Academic pediatric medical center. PARTICIPANTS: Clinical sample of 235 children aged 3 to 18 years undergoing overnight polysomnography for symptoms of SDB. OUTCOME MEASURES: History of behavioral, emotional, and academic problems and Child Behavior Checklist (CBCL) scores. RESULTS: More than half (56%) of the sample was overweight or at risk for overweight, more than one-third (36%) was classified as being short sleepers, and almost half (49%) had at least 1 additional sleep diagnosis. Forty-seven percent had a history of behavioral problems and 23% had a reported diagnosis of attention-deficit/hyperactivity disorder. There were no significant differences in CBCL scores based on any measure of SDB disease severity. Increased weight was associated with increased internalizing CBCL scores in a dose-dependent fashion (P = .003), while short sleepers were more likely to have elevated externalizing scores (P < .001). Overall, the strongest predictor of adverse behavioral outcomes was the presence of at least 1 additional sleep diagnosis (P < .001). CONCLUSIONS: The relationship between SDB and parent-reported behavioral outcomes in children is complex. In addition to SDB-related impairments, clinicians should consider the relative contributions of being overweight, insufficient sleep, and comorbid sleep disorders when assessing behavior in these children. PMID- 18391140 TI - Actigraphic and parental reports of sleep difficulties in children with attention deficit/hyperactivity disorder. AB - OBJECTIVES: To describe actigraphically detected and parent-reported sleep problems in nonmedicated children with attention-deficit/hyperactivity disorder (ADHD); to clarify whether or not comorbid oppositional defiant disorder contributes to sleep difficulties; and to compare objectively measured sleep with the parents' observations of sleep. DESIGN: Case-control study. SETTING: A child and adolescent psychiatric department of a teaching hospital. PARTICIPANTS: Two hundred six children aged 5 to 11 years, including 45 with a diagnosis of ADHD, 64 with a diagnosis of other psychiatric diagnoses (psychiatric control group), and 97 healthy control subjects (reference group). Intervention Sleep was monitored by parent-completed sleep diaries and 5 nights of actigraphy. We used a semistructured interview to diagnose psychiatric disorders according to Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) criteria. MAIN OUTCOME MEASURES: Actigraphically measured sleep variables and parent estimated sleep by diary. RESULTS: We found that children with ADHD have significantly longer sleep onset latency and a more irregular sleep pattern than the psychiatric control or healthy reference subjects. Average sleep onset latencies were 26.3 minutes in the ADHD group, 18.6 minutes in the psychiatric control group, and 13.5 minutes in the healthy reference group. There was no apparent relationship between sleep problems and comorbid oppositional defiant disorder. We found discrepancies between the objectively measured sleep variables and those reported by parents, who overestimated sleep onset latency. CONCLUSIONS: The results of this study allow us to conclude that some children with ADHD have impaired sleep that cannot be referred to comorbid oppositional defiant disorder. However, it is important to make an in-depth review of the sleep complaints, as the problem may be a product of the parents' perception rather than the child's actual experience. PMID- 18391141 TI - Parent-reported sleep problems during development and self-reported anxiety/depression, attention problems, and aggressive behavior later in life. AB - OBJECTIVE: To examine associations between sleep problems during development and subsequent emotional and behavioral difficulties. DESIGN: Prospective longitudinal study. SETTING: The Dutch province of Zuid-Holland. PARTICIPANTS: At time 1 of data collection, a representative sample of 2076 children aged 4 to 16 years participated in the study. OUTCOME MEASURES: Parents rated their children's (4-19 years old) sleep at 5 assessments by completing the Child Behavior Checklist. Participants reported on their own emotional and behavioral symptoms at a later assessment (when aged 18-32 years) by completing the Young Adult Self Report. RESULTS: After adjusting for sex, age, socioeconomic status, and parent rated scores through development for the difficulty being predicted, having any parental reports of sleeping less than others was a risk indicator of high scores on the Anxious/Depressed scale (odds ratio, 1.43; 95% confidence interval, 1.07 1.90; P = .01) and the Aggressive Behavior scale (odds ratio, 1.51; 95% confidence interval, 1.13-2.02; P = .005). There was some (albeit less robust) support for links between other reported sleep difficulties and later problems. Parental reports of sleeping more than others and nightmares were not associated with later difficulties. CONCLUSIONS: Physicians should inquire about sleep problems during child development and should be aware that some, but perhaps not others, may constitute risk indicators of later difficulties. PMID- 18391142 TI - Sleep problems in children with attention-deficit/hyperactivity disorder: prevalence and the effect on the child and family. AB - OBJECTIVES: To determine the prevalence of sleep problems in children with attention-deficit/hyperactivity disorder (ADHD) and their associations with child quality of life (QOL), daily functioning, and school attendance; caregiver mental health and work attendance; and family functioning. DESIGN: Cross-sectional survey. SETTING: Pediatric hospital outpatient clinic, private pediatricians' offices, and ADHD support groups in Victoria, Australia. PARTICIPANTS: Schoolchildren with ADHD. Main Exposure Attention-deficit/hyperactivity disorder. OUTCOME MEASURES: Primary measure was caregivers' reports of their children's sleep problems (none, mild, or moderate or severe). Secondary outcomes were (1) child QOL (Pediatric Quality of Life Inventory), daily functioning (Daily Parent Rating of Evening and Morning Behavior scale), and school attendance, (2) caregiver mental health (Depression Anxiety Stress Scale) and work attendance, and (3) family functioning (Child Health Questionnaire subscales). Caregivers also reported on how their pediatrician treated their children's sleep problems. RESULTS: Two hundred thirty-nine of 330 (74%) eligible families completed the survey. Child sleep problems were common (mild, 28.5%; moderate or severe, 44.8%). Moderate or severe sleep problems were associated with poorer child psychosocial QOL, child daily functioning, caregiver mental health, and family functioning. After adjusting for confounders, all associations held except for family impacts. Compared with children without sleep problems, those with sleep problems were more likely to miss or be late for school, and their caregivers were more likely to be late to work. Forty-five percent of caregivers reported that their pediatricians had asked about their children's sleep and, of these, 60% reported receiving treatment advice. CONCLUSIONS: Sleep problems in children with ADHD are common and associated with poorer child, caregiver, and family outcomes. Future research needs to determine whether management of sleep problems can reduce adverse outcomes. PMID- 18391143 TI - Development of sleep-wake schedules during childhood and relationship with sleep duration. AB - OBJECTIVES: To present a longitudinal overview of the sleep schedules during weekdays and the weekend in a large sample of children and to examine the sleep wake schedules in relation to nocturnal sleep duration and birth season. DESIGN: Longitudinal survey design. SETTING: Data were collected by home interviews and questionnaires. PARTICIPANTS: Data for sleep schedules at the ages of 4, 5, and 6 years on weekdays were obtained for 1112 children. Data for bedtimes on the weekend were available for 1094 children, and data for wake times on the weekend were available for 1083 children. OUTCOME MEASURES: Habitual bedtimes and wake times during weekdays and the weekend were reported by the mothers at the 3 ages. Nighttime sleep duration was calculated from bedtime to wake time. Birth characteristics came from medical records. RESULTS: As children grow up, we found later bedtimes on the weekend for a given bedtime on weekdays. Interestingly, most children slept less during the weekend compared with weekdays. No significant difference (P > .10 for all) was found in the seasonal distribution of births in any of the sleep-wake schedules. CONCLUSIONS: Despite a lack of a strong preference for sleep timing during childhood, the displacement of bedtime toward later hours during the weekend starts early in life. More important, sleep wake patterns are associated with sleep duration and later bedtimes on the weekend seem to shorten sleep duration in most children. PMID- 18391144 TI - Agreement rates between actigraphy, diary, and questionnaire for children's sleep patterns. AB - OBJECTIVES: To describe sleep-wake patterns in kindergarten children by measures derived from questionnaire, diary, and actigraphy and to report rates of agreement between methods according to Bland and Altman. DESIGN: Cross-sectional study, data from 7 nights of actigraph recordings and sleep diary and from a questionnaire. SETTING: Children studied in their homes. PARTICIPANTS: Fifty children, aged 4 to 7 years. MAIN OUTCOME MEASURES: Sleep start, sleep end, assumed sleep, actual sleep time, and nocturnal wake time derived from different methods. RESULTS: Differences between actigraphy and diary were +/- 28 minutes for sleep start, +/- 24 minutes for sleep end, and +/- 32 minutes for assumed sleep, indicating satisfactory agreement between methods, whereas for actual sleep time and nocturnal wake time, agreement rates were not sufficient (+/- 106 minutes and +/- 55 minutes, respectively). Agreement rates between actigraphy and questionnaire as well as between diary and questionnaire were insufficient for all variables. Sex and age of children and socioeconomic status did not influence the differences between methods for all variables. CONCLUSIONS: Actigraphy and diary may be interchangeably used for the assessment of sleep start, sleep end, and assumed sleep but not for nocturnal wake times. The diary is a cost-effective and valid source of information about children's sleep-schedule times, while actigraphy may provide additional information about nocturnal wake times or may be used if parents are unable to report in detail. It is insufficient to collect information by a questionnaire or an interview asking about children's normal sleep patterns. PMID- 18391145 TI - Longitudinal study of preschool sleep disturbance: the predictive role of maladaptive parental behaviors, early sleep problems, and child/mother psychological factors. AB - OBJECTIVE: To determine if maladaptive parental behaviors (at age 29-41 months) and mother/child psychological characteristics predict future sleep disturbances in 50-month-old to 6-year-old preschoolers, while controlling for early (age 5-17 months) sleep and sociodemographic factors. DESIGN: Randomized survey; children assessed annually from 5 months to 6 years of age. SETTING: Participants' homes. PARTICIPANTS: Representative sample of 987 children born in the province of Quebec, Canada, in 1997-1998. MAIN OUTCOME MEASURES: Questionnaires and interview, including responses from 7 points for 3 key dependent measures: bad dreams (BD), total sleep time (TST) less than 10 hours/night, and sleep-onset latency (SOL) of 15 minutes or more. RESULTS: Early (age 5-17 months) sleep disturbances predicted maladaptive parental behaviors (eg, mother present at sleep onset, giving food/drink after child awakens) at ages 29 and 41 months. Some parental behaviors in turn predicted future BD, TST less than 10 hours/night, and SOL of 15 minutes or more. However, most relationships did not remain significant in adjusted models that controlled for early sleep problems. Bad dreams were predicted by psychological variables (child's anxiety, mother's feeling of efficacy), as was TST (child's difficult temperament and anxiety, mother's depressive symptoms). However, SOL of 15 minutes or more was predicted by several parental behaviors even in adjusted models; cosleeping after awakenings was a risk factor while mother's presence at sleep onset was a protective factor. CONCLUSIONS: Findings support the hypothesis that maladaptive parental behaviors develop in reaction to preexisting sleep difficulties. Further, early sleep difficulties are more predictive than parental behaviors in explaining BD and foreshortened TST beginning at age 50 months. Results are interpreted in light of early emotive/physiological self-regulation problems. PMID- 18391146 TI - Pediatricians' reports of their education in ethics. AB - OBJECTIVE: To study pediatricians' assessments of the quality of their ethics education, the impact of various learning methods, and their confidence in confronting ethical dilemmas arising in pediatric practice. DESIGN: Cross sectional survey. PARTICIPANTS: Two hundred fifty physicians who completed pediatric or medicine/pediatric residency programs in 2004 were randomly selected from the American Medical Association Physician Masterfile. Evaluable responses were received from 150 of 215 eligible pediatricians (70%). RESULTS: Of 150 respondents, 44.7% rated their ethics education during residency as fair or poor. More than 80% reported that informal discussions with fellow residents and attending physicians had a moderate or major effect on their ethics education, whereas 53.3% reported that formal teaching conferences had a moderate or major impact. Most respondents (>60%) reported confidence in addressing 4 of 23 ethical challenges, a moderate proportion (40%-60%) reported confidence in addressing 8 of 23 ethical challenges, and fewer (<40%) reported confidence in addressing 11 of the ethical challenges. Areas associated with low confidence included ethics in end-of-life care and research ethics. CONCLUSIONS: Efforts are needed to augment formal and informal ethics teaching during residency. Additional studies at both the individual physician and residency program levels are needed to improve the ethics education that pediatricians-in-training receive. PMID- 18391147 TI - Secular trends in the prevalence of iron deficiency among US toddlers, 1976-2002. AB - OBJECTIVE: To examine secular trends in iron deficiency among US children 1 to 3 years old. DESIGN: Secular trend analyses of the National Health and Nutrition Examination Survey II-IV. SETTING: Large-scale national survey conducted by the National Center for Health Statistics from 1976 to 2002. PARTICIPANTS: US children 1 to 3 years old. Outcome Measure Prevalence of iron deficiency. RESULTS: Between 1976 and 2002, there was no change in iron deficiency prevalence in US toddlers. Iron deficiency prevalence remained unchanged in Hispanic and white toddlers but decreased among black toddlers. Across all 3 survey waves, racial/ethnic disparities in iron deficiency persisted between Hispanic and white toddlers, with a disparity ratio of at least 2. Iron deficiency prevalence remained high (20%-24%) in overweight toddlers, significantly higher than in those at risk for overweight (11%) and in normal weight or underweight toddlers (8%). Iron deficiency prevalence decreased from 22% to 9% in toddlers in poor households but remained unchanged in toddlers in nonpoor households (7%). In multivariable analyses, Hispanic, younger, and overweight toddlers had higher odds of iron deficiency. CONCLUSIONS: Despite the decline in iron deficiency prevalence among 1-year-old, black, and poor children, iron deficiency prevalence in US toddlers overall has not changed in the last 26 years and remains elevated in certain high-risk groups: Hispanic, younger, and overweight toddlers. Efforts to reduce the prevalence of iron deficiency in infancy and early childhood are urgently needed and should target high-risk groups. PMID- 18391148 TI - Picture of the month. Urticaria pigmentosa. PMID- 18391149 TI - Sleep difficulties and behavioral outcomes in children. PMID- 18391150 TI - Stages of a toy recall. PMID- 18391151 TI - Teaching NeuroImage: Diverse MRI signal intensities with Wernicke encephalopathy. PMID- 18391152 TI - Teaching NeuroImage: Convergence spasm associated with midbrain compression by cerebral aneurysm. PMID- 18391154 TI - Dissecting brain death: time for a new look. PMID- 18391155 TI - The time is right to improve organization of stroke care. PMID- 18391156 TI - Objective monitoring of tremor and bradykinesia during DBS surgery for Parkinson disease. AB - OBJECTIVE: High-frequency subthalamic nucleus deep brain stimulation (STN-DBS) is an established treatment for patients with advanced Parkinson disease (PD). To date, intraoperative monitoring of parkinsonian symptoms, such as tremor and bradykinesia, is largely based on subjective strategies. We conducted a pilot study to evaluate short-term intraoperative outcomes of unilateral macrostimulation of the STN-DBS in PD patients using a neuromotor symptom registration device (CATSYS 2000 System). METHODS: We studied 12 consecutive PD patients who received staged unilateral STN-DBS implants and 10 male control subjects free of neurologic deficits using a simple portable system with two sensors: a tremor pen and a touch recording plate. Results revealed excellent test-retest reliability for postural tremor in control subjects. PD patients were evaluated preoperatively during "off" state and intraoperatively for rest, postural tremor intensity, and frequency of finger tapping. Comparisons between premacrostimulation and postmacrostimulation were made using analysis of variance for repeated measures. RESULTS: Electronic rest tremor registration revealed a mean improvement of x 12.5 in tremor intensity measurements in the stimulated/contralateral side (p = 0.002). An overall x 3.8 improvement was registered on the nonstimulated/ipsilateral side. Significant improvements after STN-DBS were also recorded for postural tremor and frequency of finger tapping. CONCLUSION: Using a noninvasive, simple, and sensitive electronic recording method of intraoperative motor symptom registration, we were able to supplement short-term clinical observation by objectively quantifying the characteristics of tremor and finger tapping in response to subthalamic nucleus deep brain macrostimulation. PMID- 18391157 TI - History of depression, depressive symptoms, and medial temporal lobe atrophy and the risk of Alzheimer disease. AB - BACKGROUND: Depression may increase risk for Alzheimer disease (AD), but it is not clear whether this risk is mediated by structural brain changes. We determined whether history of depressive episodes and presence of depressive symptoms were associated with smaller hippocampal and amygdalar volumes and with increased risk for incident AD. METHODS: Within the Rotterdam Scan Study 503 persons, aged 60-90 years at baseline and without dementia, reported their history of depressive episodes. Depressive symptoms were assessed with the Center for Epidemiologic Studies Depression Scale. Volumetric assessment of the hippocampus and amygdala was performed using three-dimensional MRI. All subjects were followed for an average of 6 years for development of AD, diagnosed according to National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association criteria. RESULTS: A total of 134 subjects (26.6%) reported a history of depression (88 reported an onset <60 years and 46 a late onset). Multiple linear regression analyses did not reveal a significant association with hippocampal or amygdalar volume for any of the depression parameters. During follow-up, 33 persons developed AD. Cox regression analyses showed that subjects with early onset depression had an increased risk for AD (HR 3.76; 95% CI 1.41 to 10.06), independent of hippocampal and amygdalar volume, whereas this risk was 2.34 (95% CI 0.82 to 6.69) in subjects with a late-onset depression. Depressive symptoms at baseline were not associated with increased risk for AD. CONCLUSION: History of depression, and particularly an early onset, but not presence of depressive symptoms increased the risk for Alzheimer disease. This risk was not mediated by smaller hippocampal or amygdalar volumes. PMID- 18391158 TI - Temporal distributions of seizure occurrence from various epileptogenic regions. AB - OBJECTIVE: The aim of this study was to determine whether seizure occurrence in partial epilepsy is under the influence of circadian rhythms and rhythmic exogenous factors, and how this influence varies according to cortical brain region. For these ends, we determined and analyzed detailed temporal distributions of seizures arising from the frontal, parietal, occipital, neocortical temporal, and mesial temporal lobes. METHODS: We retrospectively analyzed intracranial EEG recordings from 131 consecutive adult subjects whose partial epilepsy was sufficiently localized for surgical resection. In all, 669 seizures were analyzed: 132 frontal, 77 parietal, 83 occipital, 217 mesial temporal, and 160 neocortical temporal. RESULTS: Seizure distribution was dependent on brain region (p < 10(-9)). Nonuniform seizure distributions were observed in the parietal (p < 10(-4)), occipital (p < 10(-7)), mesial temporal (p < 0.02), and neocortical temporal lobes (p < 0.04). Occipital and parietal seizures occurred in strong gaussian-like distributions, 180 degrees out of phase relative to each other; occipital seizure occurrence peaked between 16:00 and 19:00, whereas parietal seizures peaked between 4:00 and 7:00. Frontal lobe seizures followed a unimodal distribution, peaking between 4:00 and 7:00. Seizures from the mesial temporal lobe were distributed bimodally, with the primary peak in the late afternoon between 16:00 and 19:00 and secondary peak in the morning between 7:00 and 10:00. Neocortical temporal seizures peaked slightly before the primary peak observed in the mesial temporal lobe; however, these distributions did not differ significantly. CONCLUSIONS: Seizure occurrence in partial epilepsy is not random. Endogenous circadian rhythms and rhythmic exogenous factors likely play substantial roles in seizure occurrence. These roles vary considerably according to brain region. Frontal and parietal lobe seizures seem most likely to occur nocturnally, whereas occipital and temporal lobe seizures seem to have strong afternoon preferences. PMID- 18391159 TI - Systematic review of therapeutic interventions in human prion disease. AB - BACKGROUND: The potential threat of a large outbreak of variant Creutzfeldt-Jakob disease initiated a proliferation of research into the understanding and treatment of human prion disease. However, clinical research is at an early stage with a pressing need for objective evaluation of treatments to inform the design of future studies. METHODS: We aimed to summarize existing research on outcomes of patients with prion disease, considering any published clinical study and patient series with data on disease progression. Methods were prespecified in a protocol and studies were identified from systematic searches of multiple sources. RESULTS: One randomized trial was identified. Many studies were flawed or poorly reported, and therefore interpreted cautiously. One hundred forty published patient series revealed wide ranges in disease duration for each of the prion diseases. Thirty-three studies described the use of 14 drugs, 10 which were reported in single studies of three or fewer patients and one which was reported for two individual cases. Effects of four drugs were examined in more detail, with mixed results. The only current reliable evidence is from the single randomized trial suggesting that flupirtine may slow cognitive decline. Based on published information identified by this review, survival of most treated patients is within the ranges reported in the untreated patient series. CONCLUSIONS: Thirty years of clinical investigation of patients with prion disease has resulted in little progress in either defining or evaluating potential treatments. Disease course and treatment of all patients must be evaluated within a structured framework, preferably within randomized controlled trials. PMID- 18391160 TI - Intracerebral hemorrhage associated with use of tadalafil (Cialis). PMID- 18391161 TI - Gastrointestinal tract involvement associated with the 3243A>G mitochondrial DNA mutation. PMID- 18391162 TI - Opsoclonus-myoclonus syndrome associated with benign ovarian teratoma. PMID- 18391163 TI - Giant arteriovenous malformation presenting as pediatric task-specific dystonia. PMID- 18391164 TI - Re: Status epilepticus: an independent outcome predictor after cerebral anoxia. PMID- 18391165 TI - Re: The Stroke Outcomes and Neuroimaging of Intracranial Atherosclerosis (SONIA) trial. PMID- 18391166 TI - Nonsynaptic chemical transmission through nicotinic acetylcholine receptors. AB - This review attempts to organize the different aspects of nicotinic transmission in the context of nonsynaptic interactions. Nicotinic acetylcholine receptors (nAChRs) dominantly operate in the nonsynaptic mode in the central nervous system despite their ligand-gated ion-channel nature, which would otherwise be better suited for fast synaptic transmission. This fast form of nonsynaptic transmission, most likely unique to nAChRs, represents a new avenue in the communication platforms of the brain. Cholinergic messages received by nAChRs, arriving at a later phase following synaptic activation, can interfere with dendritic signal integration. Nicotinic transmission plays a role in both neural plasticity and cellular learning processes, as well as in long-term changes in basic activity through fast activation, desensitization of receptors, and fluctuations of the steady-state levels of ACh. ACh release can contribute to plastic changes via activation of nAChRs in neurons and therefore plays a role in learning and memory in different brain regions. Assuming that nAChRs in human subjects are ready to receive long-lasting messages from the extracellular space because of their predominantly nonsynaptic distribution, they offer an ideal target for drug therapy at low, nontoxic drug levels. PMID- 18391167 TI - CLC-0 and CFTR: chloride channels evolved from transporters. AB - CLC-0 and cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channels play important roles in Cl(-) transport across cell membranes. These two proteins belong to, respectively, the CLC and ABC transport protein families whose members encompass both ion channels and transporters. Defective function of members in these two protein families causes various hereditary human diseases. Ion channels and transporters were traditionally viewed as distinct entities in membrane transport physiology, but recent discoveries have blurred the line between these two classes of membrane transport proteins. CLC-0 and CFTR can be considered operationally as ligand-gated channels, though binding of the activating ligands appears to be coupled to an irreversible gating cycle driven by an input of free energy. High-resolution crystallographic structures of bacterial CLC proteins and ABC transporters have led us to a better understanding of the gating properties for CLC and CFTR Cl(-) channels. Furthermore, the joined force between structural and functional studies of these two protein families has offered a unique opportunity to peek into the evolutionary link between ion channels and transporters. A promising byproduct of this exercise is a deeper mechanistic insight into how different transport proteins work at a fundamental level. PMID- 18391169 TI - Ca2+-operated transcriptional networks: molecular mechanisms and in vivo models. AB - Calcium is the most universal signal used by living organisms to convey information to many different cellular processes. In this review we present well known and recently identified proteins that sense and decode the calcium signal and are key elements in the nucleus to regulate the activity of various transcriptional networks. When possible, the review also presents in vivo models in which the genes encoding these calcium sensors-transducers have been modified, to emphasize the critical role of these Ca(2+)-operated mechanisms in many physiological functions. PMID- 18391170 TI - Properties of the glomerular barrier and mechanisms of proteinuria. AB - This review focuses on the intricate properties of the glomerular barrier. Other reviews have focused on podocyte biology, mesangial cells, and the glomerular basement membrane (GBM). However, since all components of the glomerular membrane are important for its function, proteinuria will occur regardless of which layer is affected by disease. We review the properties of endothelial cells and their surface layer, the GBM, and podocytes, discuss various methods of studying glomerular permeability, and analyze data concerning the restriction of solutes by size, charge, and shape. We also review the physical principles of transport across biological or artificial membranes and various theoretical models used to predict the fluxes of solutes and water. The glomerular barrier is highly size and charge selective, in qualitative agreement with the classical studies performed 30 years ago. The small amounts of albumin filtered will be reabsorbed by the megalin-cubulin complex and degraded by the proximal tubular cells. At present, there is no unequivocal evidence for reuptake of intact albumin from urine. The cellular components are the key players in restricting solute transport, while the GBM is responsible for most of the resistance to water flow across the glomerular barrier. PMID- 18391171 TI - Regulation of actin assembly associated with protrusion and adhesion in cell migration. AB - To migrate, a cell first extends protrusions such as lamellipodia and filopodia, forms adhesions, and finally retracts its tail. The actin cytoskeleton plays a major role in this process. The first part of this review (sect. II) describes the formation of the lamellipodial and filopodial actin networks. In lamellipodia, the WASP-Arp2/3 pathways generate a branched filament array. This polarized dendritic actin array is maintained in rapid treadmilling by the concerted action of ADF, profilin, and capping proteins. In filopodia, formins catalyze the processive assembly of nonbranched actin filaments. Cell matrix adhesions mechanically couple actin filaments to the substrate to convert the treadmilling into protrusion and the actomyosin contraction into traction of the cell body and retraction of the tail. The second part of this review (sect. III) focuses on the function and the regulation of major proteins (vinculin, talin, tensin, and alpha-actinin) that control the nucleation, the binding, and the barbed-end growth of actin filaments in adhesions. PMID- 18391172 TI - Local gene expression in axons and nerve endings: the glia-neuron unit. AB - Neurons have complex and often extensively elongated processes. This unique cell morphology raises the problem of how remote neuronal territories are replenished with proteins. For a long time, axonal and presynaptic proteins were thought to be exclusively synthesized in the cell body, which delivered them to peripheral sites by axoplasmic transport. Despite this early belief, protein has been shown to be synthesized in axons and nerve terminals, substantially alleviating the trophic burden of the perikaryon. This observation raised the question of the cellular origin of the peripheral RNAs involved in protein synthesis. The synthesis of these RNAs was initially attributed to the neuron soma almost by default. However, experimental data and theoretical considerations support the alternative view that axonal and presynaptic RNAs are also transcribed in the flanking glial cells and transferred to the axon domain of mature neurons. Altogether, these data suggest that axons and nerve terminals are served by a distinct gene expression system largely independent of the neuron cell body. Such a local system would allow the neuron periphery to respond promptly to environmental stimuli. This view has the theoretical merit of extending to axons and nerve terminals the marginalized concept of a glial supply of RNA (and protein) to the neuron cell body. Most long-term plastic changes requiring de novo gene expression occur in these domains, notably in presynaptic endings, despite their intrinsic lack of transcriptional capacity. This review enlightens novel perspectives on the biology and pathobiology of the neuron by critically reviewing these issues. PMID- 18391168 TI - Cardiac remodeling in obesity. AB - The dramatic increase in the prevalence of obesity and its strong association with cardiovascular disease have resulted in unprecedented interest in understanding the effects of obesity on the cardiovascular system. A consistent, but puzzling clinical observation is that obesity confers an increased susceptibility to the development of cardiac disease, while at the same time affording protection against subsequent mortality (termed the obesity paradox). In this review we focus on evidence available from human and animal model studies and summarize the ways in which obesity can influence structure and function of the heart. We also review current hypotheses regarding mechanisms linking obesity and various aspects of cardiac remodeling. There is currently great interest in the role of adipokines, factors secreted from adipose tissue, and their role in the numerous cardiovascular complications of obesity. Here we focus on the role of leptin and the emerging promise of adiponectin as a cardioprotective agent. The challenge of understanding the association between obesity and heart failure is complicated by the multifaceted interplay between various hemodynamic, metabolic, and other physiological factors that ultimately impact the myocardium. Furthermore, the end result of obesity-associated changes in the myocardial structure and function may vary at distinct stages in the progression of remodeling, may depend on the individual pathophysiology of heart failure, and may even remain undetected for decades before clinical manifestation. Here we summarize our current knowledge of this complex yet intriguing topic. PMID- 18391173 TI - Telomeres and aging. AB - Telomeres play a central role in cell fate and aging by adjusting the cellular response to stress and growth stimulation on the basis of previous cell divisions and DNA damage. At least a few hundred nucleotides of telomere repeats must "cap" each chromosome end to avoid activation of DNA repair pathways. Repair of critically short or "uncapped" telomeres by telomerase or recombination is limited in most somatic cells and apoptosis or cellular senescence is triggered when too many "uncapped" telomeres accumulate. The chance of the latter increases as the average telomere length decreases. The average telomere length is set and maintained in cells of the germline which typically express high levels of telomerase. In somatic cells, telomere length is very heterogeneous but typically declines with age, posing a barrier to tumor growth but also contributing to loss of cells with age. Loss of (stem) cells via telomere attrition provides strong selection for abnormal and malignant cells, a process facilitated by the genome instability and aneuploidy triggered by dysfunctional telomeres. The crucial role of telomeres in cell turnover and aging is highlighted by patients with 50% of normal telomerase levels resulting from a mutation in one of the telomerase genes. Short telomeres in such patients are implicated in a variety of disorders including dyskeratosis congenita, aplastic anemia, pulmonary fibrosis, and cancer. Here the role of telomeres and telomerase in human aging and aging associated diseases is reviewed. PMID- 18391175 TI - Transcriptional paradigms in mammalian mitochondrial biogenesis and function. AB - Mitochondria contain their own genetic system and undergo a unique mode of cytoplasmic inheritance. Each organelle has multiple copies of a covalently closed circular DNA genome (mtDNA). The entire protein coding capacity of mtDNA is devoted to the synthesis of 13 essential subunits of the inner membrane complexes of the respiratory apparatus. Thus the majority of respiratory proteins and all of the other gene products necessary for the myriad mitochondrial functions are derived from nuclear genes. Transcription of mtDNA requires a small number of nucleus-encoded proteins including a single RNA polymerase (POLRMT), auxiliary factors necessary for promoter recognition (TFB1M, TFB2M) and activation (Tfam), and a termination factor (mTERF). This relatively simple system can account for the bidirectional transcription of mtDNA from divergent promoters and key termination events controlling the rRNA/mRNA ratio. Nucleomitochondrial interactions depend on the interplay between transcription factors (NRF-1, NRF-2, PPARalpha, ERRalpha, Sp1, and others) and members of the PGC-1 family of regulated coactivators (PGC-1alpha, PGC-1beta, and PRC). The transcription factors target genes that specify the respiratory chain, the mitochondrial transcription, translation and replication machinery, and protein import and assembly apparatus among others. These factors are in turn activated directly or indirectly by PGC-1 family coactivators whose differential expression is controlled by an array of environmental signals including temperature, energy deprivation, and availability of nutrients and growth factors. These transcriptional paradigms provide a basic framework for understanding the integration of mitochondrial biogenesis and function with signaling events that dictate cell- and tissue-specific energetic properties. PMID- 18391176 TI - Molecular physiology of bestrophins: multifunctional membrane proteins linked to best disease and other retinopathies. AB - This article reviews the current state of knowledge about the bestrophins, a newly identified family of proteins that can function both as Cl(-) channels and as regulators of voltage-gated Ca(2+) channels. The founding member, human bestrophin-1 (hBest1), was identified as the gene responsible for a dominantly inherited, juvenile-onset form of macular degeneration called Best vitelliform macular dystrophy. Mutations in hBest1 have also been associated with a small fraction of adult-onset macular dystrophies. It is proposed that dysfunction of bestrophin results in abnormal fluid and ion transport by the retinal pigment epithelium, resulting in a weakened interface between the retinal pigment epithelium and photoreceptors. There is compelling evidence that bestrophins are Cl(-) channels, but bestrophins remain enigmatic because it is not clear that the Cl(-) channel function can explain Best disease. In addition to functioning as a Cl(-) channel, hBest1 also is able to regulate voltage-gated Ca(2+) channels. Some bestrophins are activated by increases in intracellular Ca(2+) concentration, but whether bestrophins are the molecular counterpart of Ca(2+) activated Cl(-) channels remains in doubt. Bestrophins are also regulated by cell volume and may be a member of the volume-regulated anion channel family. PMID- 18391177 TI - Physiology of the prion protein. AB - Prion diseases are transmissible spongiform encephalopathies (TSEs), attributed to conformational conversion of the cellular prion protein (PrP(C)) into an abnormal conformer that accumulates in the brain. Understanding the pathogenesis of TSEs requires the identification of functional properties of PrP(C). Here we examine the physiological functions of PrP(C) at the systemic, cellular, and molecular level. Current data show that both the expression and the engagement of PrP(C) with a variety of ligands modulate the following: 1) functions of the nervous and immune systems, including memory and inflammatory reactions; 2) cell proliferation, differentiation, and sensitivity to programmed cell death both in the nervous and immune systems, as well as in various cell lines; 3) the activity of numerous signal transduction pathways, including cAMP/protein kinase A, mitogen-activated protein kinase, phosphatidylinositol 3-kinase/Akt pathways, as well as soluble non-receptor tyrosine kinases; and 4) trafficking of PrP(C) both laterally among distinct plasma membrane domains, and along endocytic pathways, on top of continuous, rapid recycling. A unified view of these functional properties indicates that the prion protein is a dynamic cell surface platform for the assembly of signaling modules, based on which selective interactions with many ligands and transmembrane signaling pathways translate into wide-range consequences upon both physiology and behavior. PMID- 18391174 TI - Mechanisms underlying acute protection from cardiac ischemia-reperfusion injury. AB - Mitochondria play an important role in cell death and cardioprotection. During ischemia, when ATP is progressively depleted, ion pumps cannot function resulting in a rise in calcium (Ca(2+)), which further accelerates ATP depletion. The rise in Ca(2+) during ischemia and reperfusion leads to mitochondrial Ca(2+) accumulation, particularly during reperfusion when oxygen is reintroduced. Reintroduction of oxygen allows generation of ATP; however, damage to the electron transport chain results in increased mitochondrial generation of reactive oxygen species (ROS). Mitochondrial Ca(2+) overload and increased ROS can result in opening of the mitochondrial permeability transition pore, which further compromises cellular energetics. The resultant low ATP and altered ion homeostasis result in rupture of the plasma membrane and cell death. Mitochondria have long been proposed as central players in cell death, since the mitochondria are central to synthesis of both ATP and ROS and since mitochondrial and cytosolic Ca(2+) overload are key components of cell death. Many cardioprotective mechanisms converge on the mitochondria to reduce cell death. Reducing Ca(2+) overload and reducing ROS have both been reported to reduce ischemic injury. Preconditioning activates a number of signaling pathways that reduce Ca(2+) overload and reduce activation of the mitochondrial permeability transition pore. The mitochondrial targets of cardioprotective signals are discussed in detail. PMID- 18391178 TI - Role of beta-adrenoceptor signaling in skeletal muscle: implications for muscle wasting and disease. AB - The importance of beta-adrenergic signaling in the heart has been well documented, but it is only more recently that we have begun to understand the importance of this signaling pathway in skeletal muscle. There is considerable evidence regarding the stimulation of the beta-adrenergic system with beta adrenoceptor agonists (beta-agonists). Although traditionally used for treating bronchospasm, it became apparent that some beta-agonists could increase skeletal muscle mass and decrease body fat. These so-called "repartitioning effects" proved desirable for the livestock industry trying to improve feed efficiency and meat quality. Studying beta-agonist effects on skeletal muscle has identified potential therapeutic applications for muscle wasting conditions such as sarcopenia, cancer cachexia, denervation, and neuromuscular diseases, aiming to attenuate (or potentially reverse) the muscle wasting and associated muscle weakness, and to enhance muscle growth and repair after injury. Some undesirable cardiovascular side effects of beta-agonists have so far limited their therapeutic potential. This review describes the physiological significance of beta-adrenergic signaling in skeletal muscle and examines the effects of beta agonists on skeletal muscle structure and function. In addition, we examine the proposed beneficial effects of beta-agonist administration on skeletal muscle along with some of the less desirable cardiovascular effects. Understanding beta adrenergic signaling in skeletal muscle is important for identifying new therapeutic targets and identifying novel approaches to attenuate the muscle wasting concomitant with many diseases. PMID- 18391179 TI - Dendritic excitability and synaptic plasticity. AB - Most synaptic inputs are made onto the dendritic tree. Recent work has shown that dendrites play an active role in transforming synaptic input into neuronal output and in defining the relationships between active synapses. In this review, we discuss how these dendritic properties influence the rules governing the induction of synaptic plasticity. We argue that the location of synapses in the dendritic tree, and the type of dendritic excitability associated with each synapse, play decisive roles in determining the plastic properties of that synapse. Furthermore, since the electrical properties of the dendritic tree are not static, but can be altered by neuromodulators and by synaptic activity itself, we discuss how learning rules may be dynamically shaped by tuning dendritic function. We conclude by describing how this reciprocal relationship between plasticity of dendritic excitability and synaptic plasticity has changed our view of information processing and memory storage in neuronal networks. PMID- 18391180 TI - The role of the clinical laboratory in teaching and learning family nursing skills. AB - This exploratory case study of the role of the clinical laboratory in teaching and learning family nursing skills was undertaken at the Family Nursing Unit (FNU), University of Calgary. Qualitative data were gathered from current graduate students, graduates of the Master of Nursing program, faculty members, and clients of the FNU using participant observation, in-depth interviews, and review of documents. Content analysis was used to identify key themes. The study found that the family nursing clinical laboratory provided a safe, structured learning environment where students had the opportunity to observe expert clinical practice in family nursing and undertake closely supervised practice. Graduates felt well prepared and confident in their advanced nursing practice with families. Key to the success of the clinical laboratory was the teaching faculty who were skilled clinicians in family systems nursing and who used this same theoretical orientation to guide the relational, strengths-based, and collaborative learning with students. PMID- 18391181 TI - Family management styles and ADHD: utility and treatment implications. AB - A previous study identified four family management styles (FMSs) exhibited in families with children with attention-deficit/hyperactivity disorder (ADHD) and suggested that understanding how families deal with a child's ADHD would provide additional information from which to create effective interventions. The present study used the FMS typology with a sample of children and adolescents with ADHD with the aims of demonstrating that FMSs could be reliably identified in a different clinical sample and clarifying changes in FMS that occur with treatment. All four FMSs were reliably identified in the sample and more than half of the families (56.3%) improved to a higher functioning FMS with treatment. The findings suggest that FMSs can elicit important information about family functioning and may assist clinical understanding of the child-family interaction that in turn facilitates treatment. PMID- 18391182 TI - Validity and reliability of the Chinese version of the Feetham Family Functioning Survey (FFFS). AB - A Chinese version of the Feetham Family Functioning Survey (Chinese FFFS) was developed and psychometrically tested using a sample of 317 child-rearing mothers in Hong Kong. The Chinese FFFS is a self-administered questionnaire consisting of 25 items from which an instrument discrepant score (d score) can be extracted. The results from the confirmatory factor analysis and exploratory factor analysis confirm that the Chinese FFFS has a five-factor structure based on the family ecological model, thereby affirming its construct validity. Cronbach's alpha for d scores was .91, indicating a high internal consistency. In the test-retest study of 39 mothers, the correlation coefficient for "total d score" over a 2 week period was .82, which showed high test-retest reliability. The highest discrepant scores in family functioning were at the level of the marital relationship, which requires family nurses to be skilled at assessing and intervening at this family subsystem. PMID- 18391183 TI - Sleep directly following learning benefits consolidation of spatial associative memory. AB - The last decade has brought forth convincing evidence for a role of sleep in non declarative memory. A similar function of sleep in episodic memory is supported by various correlational studies, but direct evidence is limited. Here we show that cued recall of face-location associations is significantly higher following a 12-h retention interval containing sleep than following an equally long period of waking. Furthermore, retention is significantly higher over a 24-h sleep-wake interval than over an equally long wake-sleep interval. This difference occurs because retention during sleep was significantly better when sleep followed learning directly, rather than after a day of waking. These data demonstrate a beneficial effect of sleep on memory that cannot be explained solely as a consequence of reduced interference. Rather, our findings suggest a competitive consolidation process, in which the fate of a memory depends, at least in part, on its relative stability at sleep onset: Strong memories tend to be preserved, while weaker memories erode still further. An important aspect of memory consolidation may thus result from the removal of irrelevant memory "debris." PMID- 18391184 TI - Involvement of basolateral amygdala alpha2-adrenoceptors in modulating consolidation of inhibitory avoidance memory. AB - These experiments investigated the role of the alpha(2)-adrenoceptors of the basolateral nucleus of the amygdala (BLA) in modulating the retention of inhibitory avoidance (IA). In Experiment 1, male Sprague Dawley rats implanted with bilateral cannulae in the BLA received microinfusions of a selective alpha(2)-adrenoceptor antagonist idazoxan 20 min either before or immediately after training. Retention was tested 48 h later. Idazoxan induced a dose dependent enhancement of retention performance and was more effective when administered post-training. In Experiment 2, animals received pre- or post training intra-BLA infusions of a selective alpha(2)-adrenoceptor agonist UK 14,304. The agonist induced a dose-dependent impairment of retention performance and, as with the antagonist treatments, post-training infusions were more effective. These results provide additional evidence that consolidation of inhibitory avoidance memory depends critically on prolonged activation of the noradrenergic system in the BLA and indicate that this modulatory influence is mediated, in part, by pre-synaptic alpha(2)-adrenoceptors. PMID- 18391185 TI - Differential roles for hippocampal areas CA1 and CA3 in the contextual encoding and retrieval of extinguished fear. AB - Recent studies demonstrate that context-specific memory retrieval after extinction requires the hippocampus. However, the contribution of hippocampal subfields to the context-dependent expression of extinction is not known. In the present experiments, we examined the roles of areas CA1 and CA3 of the dorsal hippocampus in the context specificity of extinction. After pairing an auditory conditional stimulus (CS) with an aversive footshock (unconditional stimulus or US), rats received extinction sessions in which the CS was presented without the US. In Experiment 1, pretraining neurotoxic lesions in either CA1 or CA3 eliminated the context dependence of extinguished fear. In Experiment 2, lesions of CA1 or CA3 were made after extinction training. In this case, only CA1 lesions impaired the context dependence of extinction. Collectively, these results reveal that both hippocampal areas CA1 and CA3 contribute to the acquisition of context dependent extinction, but that only area CA1 is required for contextual memory retrieval. PMID- 18391186 TI - Role of the somatostatin system in contextual fear memory and hippocampal synaptic plasticity. AB - Somatostatin has been implicated in various cognitive and emotional functions, but its precise role is still poorly understood. Here, we have made use of mice with somatostatin deficiency, based upon genetic invalidation or pharmacologically induced depletion, and Pavlovian fear conditioning in order to address the contribution of the somatostatin system to associative fear memory. The results demonstrate an impairment of foreground and background contextual but not tone fear conditioning in mice with targeted ablation of the somatostatin gene. These deficits were associated with a decrease in long-term potentiation in the CA1 area of the hippocampus. Both the behavioral and the electrophysiological phenotypes were mimicked in wild-type mice through application of the somatostatin-depleting substance cysteamine prior to fear training, whereas no further deficits were observed upon application in the somatostatin null mutants. These results suggest that the somatostatin system plays a critical role in the acquisition of contextual fear memory, but not tone fear learning, and further highlights the role of hippocampal synaptic plasticity for information processing concerning contextual information. PMID- 18391187 TI - Galpha(i2) inhibition of adenylate cyclase regulates presynaptic activity and unmasks cGMP-dependent long-term depression at Schaffer collateral-CA1 hippocampal synapses. AB - Cyclic AMP signaling plays a central role in regulating activity at a number of synapses in the brain. We showed previously that pairing activation of receptors that inhibit adenylate cyclase (AC) and reduce the concentration of cyclic AMP, with elevation of the concentration of cyclic GMP is sufficient to elicit a presynaptically expressed form of LTD at Schaffer collateral-CA1 synapses in the hippocampus. To directly test the role of AC inhibition and G-protein signaling in LTD at these synapses, we utilized transgenic mice that express a mutant, constitutively active inhibitory G protein, Galpha(i2), in principal neurons of the forebrain. Transgene expression of Galpha(i2) markedly enhanced LTD and impaired late-phase LTP at Schaffer collateral synapses, with no associated differences in input/output relations, paired-pulse facilitation, or NMDA receptor-gated conductances. When paired with application of a type V phosphodiesterase inhibitor to elevate the concentration of intracellular cyclic GMP, constitutively active Galpha(i2) expression converted the transient depression normally caused by this treatment to an LTD that persisted after the drug was washed out. Moreover, this effect could be mimicked in control slices by pairing type V phosphodiesterase inhibitor application with application of a PKA inhibitor. Electrophysiological recordings of spontaneous excitatory postsynaptic currents and two-photon visualization of vesicular release using FM1-43 revealed that constitutively active Galpha(i2) tonically reduced basal release probability from the rapidly recycling vesicle pool of Schaffer collateral terminals. Our findings support the hypothesis that inhibitory G-protein signaling acts presynaptically to regulate release, and, when paired with elevations in the concentration of cyclic GMP, converts a transient cyclic GMP-induced depression into a long-lasting decrease in release. PMID- 18391189 TI - Alloreactivity and association of human natural killer cells with the major histocompatibility complex. PMID- 18391188 TI - Acute predator stress impairs the consolidation and retrieval of hippocampus dependent memory in male and female rats. AB - We have studied the effects of an acute predator stress experience on spatial learning and memory in adult male and female Sprague-Dawley rats. All rats were trained to learn the location of a hidden escape platform in the radial-arm water maze (RAWM), a hippocampus-dependent spatial memory task. In the control (non stress) condition, female rats were superior to the males in the accuracy and consistency of their spatial memory performance tested over multiple days of training. In the stress condition, rats were exposed to the cat for 30 min immediately before or after learning, or before the 24-h memory test. Predator stress dramatically increased corticosterone levels in males and females, with females exhibiting greater baseline and stress-evoked responses than males. Despite these sex differences in the overall magnitudes of corticosterone levels, there were significant sex-independent correlations involving basal and stress evoked corticosterone levels, and memory performance. Most importantly, predator stress impaired short-term memory, as well as processes involved in memory consolidation and retrieval, in male and female rats. Overall, we have found that an intense, ethologically relevant stressor produced a largely equivalent impairment of memory in male and female rats, and sex-independent corticosterone memory correlations. These findings may provide insight into commonalities in how traumatic stress affects the brain and memory in men and women. PMID- 18391190 TI - Quality control of overexpressed membrane proteins. AB - Overexpression of membrane proteins in Escherichia coli frequently leads to the formation of aggregates or inclusion bodies, which is undesirable for most studies. Ideally, one would like to optimize the expression conditions by monitoring simultaneously and rapidly both the amounts of properly folded and aggregated membrane protein, a requirement not met by any of the currently available methods. Here, we describe a simple gel-based approach with green fluorescent protein as folding indicator to detect well folded and aggregated proteins simultaneously. The method allows for rapid screening and, importantly, pinpointing the most likely bottlenecks in protein production. PMID- 18391191 TI - Protein kinase CK2 as an ectokinase: the role of the regulatory CK2beta subunit. AB - Protein kinase CK2 (also known as casein kinase 2) is present in the cytoplasm, nuclei, and several other organelles. In addition, this enzyme has been found bound to the external side of the cell membrane where it acts as an ectokinase phosphorylating several extracellular proteins. Previous experiments with transfection of HEK-293T cells demonstrated that expression of both subunits, CK2alpha (catalytic) and CK2beta (regulatory), was necessary for the appearance of the ectopic enzyme as an ectokinase. In this work, using deletion and point mutations of CK2beta, it was possible to demonstrate that the region between amino acids 20 and 33 was necessary for the export of the enzyme as an ectokinase. Phenylalanines 21 and 22 and acidic residues in positions 26-28 are involved in the structural aspects that are required for export. However, the region encompassing amino acids 20-33 of CK2beta is not sufficient to make the carboxyl half of this subunit functional in bringing CK2 to the ectokinase locus. In cells transfected with only CK2beta, it was demonstrated that 3-4% of the subunit is exported to the cell medium, but the subunit is not bound to the external membrane. PMID- 18391192 TI - Decreased corticolimbic allopregnanolone expression during social isolation enhances contextual fear: A model relevant for posttraumatic stress disorder. AB - Mice subjected to social isolation (3-4 weeks) exhibit enhanced contextual fear responses and impaired fear extinction. These responses are time-related to a decrease of 5alpha-reductase type I (5alpha-RI) mRNA expression and allopregnanolone (Allo) levels in selected neurons of the medial prefrontal cortex, hippocampus, and basolateral amygdala. Of note, the cued fear response was not different between group housed and socially isolated mice. In socially isolated mice, S-norfluoxetine, a selective brain steroidogenic stimulant (SBSS), in doses (0.45-1.8 mumol/kg) that increase brain Allo levels but fail to inhibit serotonin reuptake, greatly attenuates enhanced contextual fear response. SKF 105,111 (a potent 5alpha-RI inhibitor) decreases corticolimbic Allo levels and enhances the contextual fear response in group housed mice, which suggests that social isolation alters emotional responses by reducing the positive allosteric modulation of Allo at GABA(A) receptors in corticolimbic circuits. Thus, these procedures model emotional hyperreactivity, including enhanced contextual fear and impaired contextual fear extinction, which also is observed in posttraumatic stress disorder (PTSD) patients. A recent clinical study reported that cerebrospinal fluid Allo levels also are down-regulated in PTSD patients and correlate negatively with PTSD symptoms and negative mood. Thus, protracted social isolation of mice combined with tests of fear conditioning may be a suitable model to study emotional behavioral components associated with neurochemical alterations relating to PTSD. Importantly, drugs like SBSSs, which rapidly increase corticolimbic Allo levels, normalize the exaggerated contextual fear responses resulting from social isolation, suggesting that selective activation of neurosteroidogenesis may be useful in PTSD therapy. PMID- 18391193 TI - Enzymatic mechanism and product specificity of SET-domain protein lysine methyltransferases. AB - Molecular dynamics and hybrid quantum mechanics/molecular mechanics have been used to investigate the mechanisms of (+)AdoMet methylation of protein-Lys-NH(2) catalyzed by the lysine methyltransferase enzymes: histone lysine monomethyltransferase SET7/9, Rubisco large-subunit dimethyltransferase, viral histone lysine trimethyltransferase, and the Tyr245Phe mutation of SET7/9. At neutrality in aqueous solution, primary amines are protonated. The enzyme reacts with Lys-NH(3)(+) and (+)AdoMet species to provide an Enz.Lys-NH(3)(+).(+)AdoMet complex. The close positioning of two positive charges lowers the pK(a) of the Lys-NH(3)(+) entity, a water channel appears, and the proton escapes to the aqueous solvent; then the reaction Enz.Lys-NH(2).(+)AdoMet --> Enz.Lys N(Me)H(2)(+).AdoHcy occurs. Repeat of the sequence provides dimethylated lysine, and another repeat yields a trimethylated lysine. The sequence is halted at monomethylation when the conformation of the Enz.Lys-N(Me)H(2)(+).(+)AdoMet has the methyl positioned to block formation of a water channel. The sequence of reactions stops at dimethylation if the conformation of Enz.Lys N(Me)(2)H(+).(+)AdoMet has a methyl in position, which forbids the formation of the water channel. PMID- 18391194 TI - A structural-functional basis for dyslexia in the cortex of Chinese readers. AB - Developmental dyslexia is a neurobiologically based disorder that affects approximately 5-17% of school children and is characterized by a severe impairment in reading skill acquisition. For readers of alphabetic (e.g., English) languages, recent neuroimaging studies have demonstrated that dyslexia is associated with weak reading-related activity in left temporoparietal and occipitotemporal regions, and this activity difference may reflect reductions in gray matter volume in these areas. Here, we find different structural and functional abnormalities in dyslexic readers of Chinese, a nonalphabetic language. Compared with normally developing controls, children with impaired reading in logographic Chinese exhibited reduced gray matter volume in a left middle frontal gyrus region previously shown to be important for Chinese reading and writing. Using functional MRI to study language-related activation of cortical regions in dyslexics, we found reduced activation in this same left middle frontal gyrus region in Chinese dyslexics versus controls, and there was a significant correlation between gray matter volume and activation in the language task in this same area. By contrast, Chinese dyslexics did not show functional or structural (i.e., volumetric gray matter) differences from normal subjects in the more posterior brain systems that have been shown to be abnormal in alphabetic language dyslexics. The results suggest that the structural and functional basis for dyslexia varies between alphabetic and nonalphabetic languages. PMID- 18391195 TI - RNA landscape of evolution for optimal exon and intron discrimination. AB - Accurate pre-mRNA splicing requires primary splicing signals, including the splice sites, a polypyrimidine tract, and a branch site, other splicing regulatory elements (SREs). The SREs include exonic splicing enhancers (ESEs), exonic splicing silencers (ESSs), intronic splicing enhancers (ISEs), and intronic splicing silencers (ISSs), which are typically located near the splice sites. However, it is unclear to what extent splicing-driven selective pressure constrains exonic and intronic sequences, especially those distant from the splice sites. Here, we studied the distribution of SREs in human genes in terms of DNA strand-asymmetry patterns. Under a neutral evolution model, each mononucleotide or oligonucleotide should have a symmetric (Chargaff's second parity rule), or weakly asymmetric yet uniform, distribution throughout a pre mRNA transcript. However, we found that large sets of unbiased, experimentally determined SREs show a distinct strand-asymmetry pattern that is inconsistent with the neutral evolution model, and reflects their functional roles in splicing. ESEs are selected in exons and depleted in introns and vice versa for ESSs. Surprisingly, this trend extends into deep intronic sequences, accounting for one third of the genome. Selection is detectable even at the mononucleotide level, so that the asymmetric base compositions of exons and introns are predictive of ESEs and ESSs. We developed a method that effectively predicts SREs based on strand asymmetry, expanding the current catalog of SREs. Our results suggest that human genes have been optimized for exon and intron discrimination through an RNA landscape shaped during evolution. PMID- 18391196 TI - Neurons derived from reprogrammed fibroblasts functionally integrate into the fetal brain and improve symptoms of rats with Parkinson's disease. AB - The long-term goal of nuclear transfer or alternative reprogramming approaches is to create patient-specific donor cells for transplantation therapy, avoiding immunorejection, a major complication in current transplantation medicine. It was recently shown that the four transcription factors Oct4, Sox2, Klf4, and c-Myc induce pluripotency in mouse fibroblasts. However, the therapeutic potential of induced pluripotent stem (iPS) cells for neural cell replacement strategies remained unexplored. Here, we show that iPS cells can be efficiently differentiated into neural precursor cells, giving rise to neuronal and glial cell types in culture. Upon transplantation into the fetal mouse brain, the cells migrate into various brain regions and differentiate into glia and neurons, including glutamatergic, GABAergic, and catecholaminergic subtypes. Electrophysiological recordings and morphological analysis demonstrated that the grafted neurons had mature neuronal activity and were functionally integrated in the host brain. Furthermore, iPS cells were induced to differentiate into dopamine neurons of midbrain character and were able to improve behavior in a rat model of Parkinson's disease upon transplantation into the adult brain. We minimized the risk of tumor formation from the grafted cells by separating contaminating pluripotent cells and committed neural cells using fluorescence activated cell sorting. Our results demonstrate the therapeutic potential of directly reprogrammed fibroblasts for neuronal cell replacement in the animal model. PMID- 18391197 TI - TFIID component TAF7 functionally interacts with both TFIIH and P-TEFb. AB - Transcription consists of a series of highly regulated steps: assembly of the preinitiation complex (PIC) at the promoter, initiation, elongation, and termination. PIC assembly is nucleated by TFIID, a complex composed of the TATA binding protein (TBP) and a series of TBP-associated factors (TAFs). One component, TAF7, is incorporated in the PIC through its interaction with TFIID but is released from TFIID upon transcription initiation. We now report that TAF7 interacts with the transcription factors, TFIIH and P-TEFb, resulting in the inhibition of their Pol II CTD kinase activities. Importantly, in in vitro transcription reactions, TAF7 inhibits steps after PIC assembly and formation of the first phosphodiester bonds. Further, in vivo TAF7 coelongates with P-TEFb and Pol II downstream of the promoter. We propose a model in which TAF7 contributes to the regulation of the transition from PIC assembly to initiation and elongation. PMID- 18391198 TI - Language universals in human brains. AB - Do speakers know universal restrictions on linguistic elements that are absent from their language? We report an experimental test of this question. Our case study concerns the universal restrictions on initial consonant sequences, onset clusters (e.g., bl in block). Across languages, certain onset clusters (e.g., lb) are dispreferred (e.g., systematically under-represented) relative to others (e.g., bl). We demonstrate such preferences among Korean speakers, whose language lacks initial C(1)C(2) clusters altogether. Our demonstration exploits speakers' well known tendency to misperceive ill-formed clusters. We show that universally dispreferred onset clusters are more frequently misperceived than universally preferred ones, indicating that Korean speakers consider the former cluster-type more ill-formed. The misperception of universally ill-formed clusters is unlikely to be due to a simple auditory failure. Likewise, the aversion of universally dispreferred onsets by Korean speakers is not explained by English proficiency or by several phonetic and phonological properties of Korean. We conclude that language universals are neither relics of language change nor are they artifacts of generic limitations on auditory perception and motor control-they reflect universal linguistic knowledge, active in speakers' brains. PMID- 18391199 TI - Complete genome of the uncultured Termite Group 1 bacteria in a single host protist cell. AB - Termites harbor a symbiotic gut microbial community that is responsible for their ability to thrive on recalcitrant plant matter. The community comprises diverse microorganisms, most of which are as yet uncultivable; the detailed symbiotic mechanism remains unclear. Here, we present the first complete genome sequence of a termite gut symbiont-an uncultured bacterium named Rs-D17 belonging to the candidate phylum Termite Group 1 (TG1). TG1 is a dominant group in termite guts, found as intracellular symbionts of various cellulolytic protists, without any physiological information. To acquire the complete genome sequence, we collected Rs-D17 cells from only a single host protist cell to minimize their genomic variation and performed isothermal whole-genome amplification. This strategy enabled us to reconstruct a circular chromosome (1,125,857 bp) encoding 761 putative protein-coding genes. The genome additionally contains 121 pseudogenes assigned to categories, such as cell wall biosynthesis, regulators, transporters, and defense mechanisms. Despite its apparent reductive evolution, the ability to synthesize 15 amino acids and various cofactors is retained, some of these genes having been duplicated. Considering that diverse termite-gut protists harbor TG1 bacteria, we suggest that this bacterial group plays a key role in the gut symbiotic system by stably supplying essential nitrogenous compounds deficient in lignocelluloses to their host protists and the termites. Our results provide a breakthrough to clarify the functions of and the interactions among the individual members of this multilayered symbiotic complex. PMID- 18391200 TI - Structure of tumor suppressor p53 and its intrinsically disordered N-terminal transactivation domain. AB - Proteins with intrinsically disordered domains are implicated in a vast range of biological processes, especially in cell signaling and regulation. Having solved the quaternary structure of the folded domains in the tumor suppressor p53 by a multidisciplinary approach, we have now determined the average ensemble structure of the intrinsically disordered N-terminal transactivation domain (TAD) by using residual dipolar couplings (RDCs) from NMR spectroscopy and small-angle x-ray scattering (SAXS). Remarkably, not only were we able to measure RDCs of the isolated TAD, but we were also able to do so for the TAD in both the full-length tetrameric p53 protein and in its complex with a specific DNA response element. We determined the orientation of the TAD ensemble relative to the core domain, found that the TAD was stiffer in the proline-rich region (residues 64-92), which has a tendency to adopt a polyproline II (PPII) structure, and projected the TAD away from the core. We located the TAD in SAXS experiments on a complex between tetrameric p53 and four Taz2 domains that bind tightly to the TAD (residues 1-57) and acted as "reporters." The p53-Taz2 complex was an extended cross-shaped structure. The quality of the SAXS data enabled us to model the disordered termini and the folded domains in the complex with DNA. The core domains enveloped the response element in the center of the molecule, with the Taz2-bound TADs projecting outward from the core. PMID- 18391201 TI - Incorrect nucleotide insertion at the active site of a G:A mismatch catalyzed by DNA polymerase beta. AB - Based on a recent ternary complex crystal structure of human DNA polymerase beta with a G:A mismatch in the active site, we carried out a theoretical investigation of the catalytic mechanism of incorrect nucleotide incorporation using molecular dynamics simulation, quantum mechanics, combined quantum mechanics, and molecular mechanics methods. A two-stage mechanism is proposed with a nonreactive active-site structural rearrangement prechemistry step occurring before the nucleotidyl transfer reaction. The free energy required for formation of the prechemistry state is found to be the major factor contributing to the decrease in the rate of incorrect nucleotide incorporation compared with correct insertion and therefore to fidelity enhancement. Hence, the transition state and reaction barrier for phosphodiester bond formation after the prechemistry state are similar to that for correct insertion reaction. Key residues that provide electrostatic stabilization of the transition state are identified. PMID- 18391202 TI - Unregulated smooth-muscle myosin in human intestinal neoplasia. AB - A recent study described a recessive ATPase activating germ-line mutation in smooth-muscle myosin (smmhc/myh11) underlying the zebrafish meltdown (mlt) phenotype. The mlt zebrafish develops intestinal abnormalities reminiscent of human Peutz-Jeghers syndrome (PJS) and juvenile polyposis (JP). To examine the role of MYH11 in human intestinal neoplasia, we searched for MYH11 mutations in patients with colorectal cancer (CRC), PJS and JP. We found somatic protein elongating frameshift mutations in 55% of CRCs displaying microsatellite instability and in the germ-line of one individual with PJS. Additionally, two somatic missense mutations were found in one microsatellite stable CRC. These two missense mutations, R501L and K1044N, and the frameshift mutations were functionally evaluated. All mutations resulted in unregulated molecules displaying constitutive motor activity, similar to the mutant myosin underlying mlt. Thus, MYH11 mutations appear to contribute also to human intestinal neoplasia. Unregulated MYH11 may affect the cellular energy balance or disturb cell lineage decisions in tumor progenitor cells. These data challenge our view on MYH11 as a passive differentiation marker functioning in muscle contraction and add to our understanding of intestinal neoplasia. PMID- 18391203 TI - EBV-encoded EBNA-6 binds and targets MRS18-2 to the nucleus, resulting in the disruption of pRb-E2F1 complexes. AB - Epstein-Barr virus (EBV), like other DNA tumor viruses, induces an S-phase in the natural host cell, the human B lymphocyte. This is linked with blast transformation. It is believed that the EBV-encoded nuclear antigen 6 (EBNA-6) is involved in the regulation of cell cycle entry. However, the possible mechanism of this regulation is not approached. In our current study, we found that EBNA-6 binds to a MRPS18-2 protein, and targets it to the nucleus. We found that MRPS18 2 binds to both hypo- and hyperphosphorylated forms of Rb protein specifically. This binding targets the small pocket of pRb, which is a site of interaction with E2F1. The MRPS18-2 competes with the binding of E2F1 to pRb, thereby raising the level of free E2F1. Our experimental data suggest that EBNA-6 may play a major role in the entry of EBV infected B cells into the S phase by binding to and raising the level of nuclear MRPS18-2, protein. This would inhibit pRb binding to E2F1 competitively and lift the block preventing S-phase entry. PMID- 18391204 TI - A polycatenated DNA scaffold for the one-step assembly of hierarchical nanostructures. AB - A unique DNA scaffold was prepared for the one-step self-assembly of hierarchical nanostructures onto which multiple proteins or nanoparticles are positioned on a single template with precise relative spatial orientation. The architecture is a topologically complex ladder-shaped polycatenane in which the "rungs" of the ladder are used to bring together the individual rings of the mechanically interlocked structure, and the "rails" are available for hierarchical assembly, whose effectiveness has been demonstrated with proteins, complementary DNA, and gold nanoparticles. The ability of this template to form from linear monomers and simultaneously bind two proteins was demonstrated by chemical force microscopy, transmission electron microscopy, and confocal fluorescence microscopy. Finally, fluorescence resonance energy transfer between adjacent fluorophores confirmed the programmed spatial arrangement between two different nanomaterials. DNA templates that bring together multiple nanostructures with precise spatial control have applications in catalysis, biosensing, and nanomaterials design. PMID- 18391205 TI - An insulin-like peptide regulates egg maturation and metabolism in the mosquito Aedes aegypti. AB - Ingestion of vertebrate blood is essential for egg maturation and transmission of disease-causing parasites by female mosquitoes. Prior studies with the yellow fever mosquito, Aedes aegypti, indicated blood feeding stimulates egg production by triggering the release of hormones from medial neurosecretory cells in the mosquito brain. The ability of bovine insulin to stimulate a similar response further suggested this trigger is an endogenous insulin-like peptide (ILP). A. aegypti encodes eight predicted ILPs. Here, we report that synthetic ILP3 dose dependently stimulated yolk uptake by oocytes and ecdysteroid production by the ovaries at lower concentrations than bovine insulin. ILP3 also exhibited metabolic activity by elevating carbohydrate and lipid storage. Binding studies using ovary membranes indicated that ILP3 had an IC(50) value of 5.9 nM that was poorly competed by bovine insulin. Autoradiography and immunoblotting studies suggested that ILP3 binds the mosquito insulin receptor (MIR), whereas loss-of function experiments showed that ILP3 activity requires MIR expression. Overall, our results identify ILP3 as a critical regulator of egg production by A. aegypti. PMID- 18391206 TI - Hybrid alphavirus-rhabdovirus propagating replicon particles are versatile and potent vaccine vectors. AB - Self-propagating, infectious, virus-like particles are generated in animal cell lines transfected with a Semliki Forest virus RNA replicon encoding a single viral structural protein, the vesicular stomatitis virus (VSV) glycoprotein. We show here that these infectious particles, which we call propagating replicons, are potent inducers of neutralizing antibody in animals yet are nonpathogenic. Mice vaccinated with a single dose of the particles generated high titers of VSV neutralizing antibody and were protected from a subsequent lethal challenge with VSV. Induction of antibody required RNA replication. We also report that additional genes (including an HIV-1 envelope protein gene) expressed from the propagating replicons induced strong cellular immune responses to the corresponding proteins after a single inoculation. Our studies reveal the potential of these particles as simple and safe vaccine vectors inducing strong humoral and cellular immune responses. PMID- 18391207 TI - Magnesium deficiency accelerates cellular senescence in cultured human fibroblasts. AB - Magnesium inadequacy affects more than half of the U.S. population and is associated with increased risk for many age-related diseases, yet the underlying mechanisms are unknown. Altered cellular physiology has been demonstrated after acute exposure to severe magnesium deficiency, but few reports have addressed the consequences of long-term exposure to moderate magnesium deficiency in human cells. Therefore, IMR-90 human fibroblasts were continuously cultured in magnesium-deficient conditions to determine the long-term effects on the cells. These fibroblasts did not demonstrate differences in cellular viability or plating efficiency but did exhibit a decreased replicative lifespan in populations cultured in magnesium-deficient compared with standard media conditions, both at ambient (20% O(2)) and physiological (5% O(2)) oxygen tension. The growth rates for immortalized IMR-90 fibroblasts were not affected under the same conditions. IMR-90 fibroblast populations cultured in magnesium deficient conditions had increased senescence-associated beta-galactosidase activity and increased p16(INK4a) and p21(WAF1) protein expression compared with cultures from standard media conditions. Telomere attrition was also accelerated in cell populations from magnesium-deficient cultures. Thus, the long-term consequence of inadequate magnesium availability in human fibroblast cultures was accelerated cellular senescence, which may be a mechanism through which chronic magnesium inadequacy could promote or exacerbate age-related disease. PMID- 18391208 TI - Probing microscopic origins of confined subdiffusion by first-passage observables. AB - Subdiffusive motion of tracer particles in complex crowded environments, such as biological cells, has been shown to be widespread. This deviation from Brownian motion is usually characterized by a sublinear time dependence of the mean square displacement (MSD). However, subdiffusive behavior can stem from different microscopic scenarios that cannot be identified solely by the MSD data. In this article we present a theoretical framework that permits the analytical calculation of first-passage observables (mean first-passage times, splitting probabilities, and occupation times distributions) in disordered media in any dimensions. This analysis is applied to two representative microscopic models of subdiffusion: continuous-time random walks with heavy tailed waiting times and diffusion on fractals. Our results show that first-passage observables provide tools to unambiguously discriminate between the two possible microscopic scenarios of subdiffusion. Moreover, we suggest experiments based on first passage observables that could help in determining the origin of subdiffusion in complex media, such as living cells, and discuss the implications of anomalous transport to reaction kinetics in cells. PMID- 18391209 TI - Adenovirus serotype 5 hexon is critical for virus infection of hepatocytes in vivo. AB - Human species C adenovirus serotype 5 (Ad5) is the most common viral vector used in clinical studies worldwide. Ad5 vectors infect liver cells in vivo with high efficiency via a poorly defined mechanism, which involves virus binding to vitamin K-dependent blood coagulation factors. Here, we report that the major Ad5 capsid protein, hexon, binds human coagulation factor X (FX) with an affinity of 229 pM. This affinity is 40-fold stronger than the reported affinity of Ad5 fiber for the cellular receptor coxsackievirus and adenovirus receptor, CAR. Cryoelectron microscopy and single-particle image reconstruction revealed that the FX attachment site is localized to the central depression at the top of the hexon trimer. Hexon-mutated virus bearing a large insertion in hexon showed markedly reduced FX binding in vitro and failed to deliver a transgene to hepatocytes in vivo. This study describes the mechanism of FX binding to Ad5 and demonstrates the critical role of hexon for virus infection of hepatocytes in vivo. PMID- 18391210 TI - Spectral completion of partially masked sounds. AB - Natural environments typically contain multiple sound sources. The sounds from these sources frequently overlap in time and often mask each other. Masking could potentially distort the representation of a sound's spectrum, altering its timbre and impairing object recognition. Here, we report that the auditory system partially corrects for the effects of masking in such situations, by using the audible, unmasked portions of an object's spectrum to fill in the inaudible portions. This spectral completion mechanism may help to achieve perceptual constancy and thus aid object recognition in complex auditory scenes. PMID- 18391211 TI - Small-molecule agonists and antagonists of F-box protein-substrate interactions in auxin perception and signaling. AB - The regulation of gene expression by the hormone auxin is a crucial mechanism in plant development. We have shown that the Arabidopsis F-box protein TIR1 is a receptor for auxin, and our recent structural work has revealed the molecular mechanism of auxin perception. TIR1 is the substrate receptor of the ubiquitin ligase complex SCF(TIR1). Auxin binding enhances the interaction between TIR1 and its substrates, the Aux/IAA repressors, thereby promoting the ubiquitination and degradation of Aux/IAAs, altering the expression of hundreds of genes. TIR1 is the prototype of a new class of hormone receptor and the first example of an SCF ubiquitin-ligase modulated by a small molecule. Here, we describe the design, synthesis, and characterization of a series of auxin agonists and antagonists. We show these molecules are specific to TIR1-mediated events in Arabidopsis, and their mode of action in binding to TIR1 is confirmed by x-ray crystallographic analysis. Further, we demonstrate the utility of these probes for the analysis of TIR1-mediated auxin signaling in the moss Physcomitrella patens. Our work not only provides a useful tool for plant chemical biology but also demonstrates an example of a specific small-molecule inhibitor of F-box protein-substrate recruitment. Substrate recognition and subsequent ubiquitination by SCF-type ubiquitin ligases are central to many cellular processes in eukaryotes, and ubiquitin-ligase function is affected in several human diseases. Our work supports the idea that it may be possible to design small-molecule agents to modulate ubiquitin-ligase function therapeutically. PMID- 18391212 TI - Identification of a potent synthetic FXR agonist with an unexpected mode of binding and activation. AB - The farnesoid X receptor (FXR), a member of the nuclear hormone receptor family, plays important roles in the regulation of bile acid and cholesterol homeostasis, glucose metabolism, and insulin sensitivity. There is intense interest in understanding the mechanisms of FXR regulation and in developing pharmaceutically suitable synthetic FXR ligands that might be used to treat metabolic syndrome. We report here the identification of a potent FXR agonist (MFA-1) and the elucidation of the structure of this ligand in ternary complex with the human receptor and a coactivator peptide fragment using x-ray crystallography at 1.9-A resolution. The steroid ring system of MFA-1 binds with its D ring-facing helix 12 (AF-2) in a manner reminiscent of hormone binding to classical steroid hormone receptors and the reverse of the pose adopted by naturally occurring bile acids when bound to FXR. This binding mode appears to be driven by the presence of a carboxylate on MFA-1 that is situated to make a salt-bridge interaction with an arginine residue in the FXR-binding pocket that is normally used to neutralize bound bile acids. Receptor activation by MFA-1 differs from that by bile acids in that it relies on direct interactions between the ligand and residues in helices 11 and 12 and only indirectly involves a protonated histidine that is part of the activation trigger. The structure of the FXR:MFA-1 complex differs significantly from that of the complex with a structurally distinct agonist, fexaramine, highlighting the inherent plasticity of the receptor. PMID- 18391213 TI - Human cremation in Mexico 3,000 years ago. AB - Mixtec nobles are depicted in codices and other proto-historic documentation taking part in funerary rites involving cremation. The time depth for this practice was unknown, but excavations at the early village site of Tayata, in the southern state of Oaxaca, Mexico, recovered undisturbed cremation burials in contexts dating from the eleventh century B.C. These are the earliest examples of a burial practice that in later times was reserved for Mixtec kings and Aztec emperors. This article describes the burial contexts and human remains, linking Formative period archaeology with ethnohistorical descriptions of Mixtec mortuary practices. The use of cremation to mark elevated social status among the Mixtec was established by 3,000 years ago, when hereditary differences in rank were first emerging across Mesoamerica. PMID- 18391214 TI - Structure of human monoamine oxidase A at 2.2-A resolution: the control of opening the entry for substrates/inhibitors. AB - The mitochondrial outer membrane-anchored monoamine oxidase (MAO) is a biochemically important flavoenzyme that catalyzes the deamination of biogenic and xenobiotic amines. Its two subtypes, MAOA and MAOB, are linked to several psychiatric disorders and therefore are interesting targets for drug design. To understand the relationship between structure and function of this enzyme, we extended our previous low-resolution rat MAOA structure to the high-resolution wild-type and G110A mutant human MAOA structures at 2.2 and 2.17 A, respectively. The high-resolution MAOA structures are similar to those of rat MAOA and human MAOB, but different from the known structure of human MAOA [De Colibus L, et al. (2005) Proc Natl Acad Sci USA 102:12684-12689], specifically regarding residues 108-118 and 210-216, which surround the substrate/inhibitor cavity. The results confirm that the inhibitor selectivity of MAOA and MAOB is caused by the structural differences arising from Ile-335 in MAOA vs. Tyr-326 in MAOB. The structures exhibit a C-terminal transmembrane helix with clear electron density, as is also seen in rat MAOA. Mutations on one residue of loop 108-118, G110, which is far from the active center but close to the membrane surface, cause the solubilized enzyme to undergo a dramatic drop in activity, but have less effect when the enzyme is anchored in the membrane. These results suggest that the flexibility of loop 108-118, facilitated by anchoring the enzyme into the membrane, is essential for controlling substrate access to the active site. We report on the observation of the structure-function relationship between a transmembrane helical anchor and an extra-membrane domain. PMID- 18391215 TI - Calcium sensitivity of the cross-bridge cycle of Myo1c, the adaptation motor in the inner ear. AB - The class I myosin Myo1c is a mediator of adaptation of mechanoelectrical transduction in the stereocilia of the inner ear. Adaptation, which is strongly affected by Ca(2+), permits hair cells under prolonged stimuli to remain sensitive to new stimuli. Using a Myo1c fragment (motor domain and one IQ domain with associated calmodulin), with biochemical and kinetic properties similar to those of the native molecule, we have performed a thorough analysis of the biochemical cross-bridge cycle. We show that, although the steady-state ATPase activity shows little calcium sensitivity, individual molecular events of the cross-bridge cycle are calcium-sensitive. Of significance is a 7-fold inhibition of the ATP hydrolysis step and a 10-fold acceleration of ADP release in calcium. These changes result in an acceleration of detachment of the cross-bridge and a lengthening of the lifetime of the detached M-ATP state. These data support a model in which slipping adaptation, which reduces tip-link tension and allows the transduction channels to close after an excitatory stimulus, is mediated by Myo1c and modulated by the calcium transient. PMID- 18391216 TI - Temperature and pressure dependence of protein stability: the engineered fluorescein-binding lipocalin FluA shows an elliptic phase diagram. AB - We have measured the equilibrium constant for the denaturation transition of the engineered fluorescein-binding lipocalin FluA as a function of pressure and temperature, taking advantage of the fact that the ligand's fluorescence is almost fully quenched when complexed with the folded protein, but reversibly reappears on denaturation. From the equilibrium constant as a function of pressure and temperature all of the involved thermodynamic parameters of protein folding, in particular the changes in entropy and volume, compressibility, thermal expansion, and specific heat, were deduced in a global fitting procedure. Assuming that these parameters are independent of temperature and pressure, we can demonstrate from the ratio of Deltabeta, Deltaalpha(2), DeltaC(p) that the phase diagram of protein folding assumes an elliptic shape. Furthermore, we can show that the thermodynamic condition for such an elliptic phase diagram is related to the degree of correlation between the fluctuations of the changes in volume and enthalpy at the phase boundary. For the protein investigated this correlation is low, as generally expected for highly degenerate systems. Our study suggests that the elliptic phase diagram is a consequence of the inherent conformational disorder of proteins and that it may be viewed as the thermodynamic manifestation of the high degeneracy of conformational energies that is characteristic for this class of macromolecules. PMID- 18391217 TI - Cytidine deamination induced HIV-1 drug resistance. AB - The HIV-1 Vif protein is essential for overcoming the antiviral activity of DNA editing apolipoprotein B mRNA editing enzyme, catalytic polypeptide 3 (APOBEC3) cytidine deaminases. We show that naturally occurring HIV-1 Vif point mutants with suboptimal anti-APOBEC3G activity induce the appearance of proviruses with lamivudine (3TC) drug resistance-associated mutations before any drug exposure. These mutations, ensuing from cytidine deamination events, were detected in >40% of proviruses with partially defective Vif mutants. Transfer of drug resistance from hypermutated proviruses via recombination allowed for 3TC escape under culture conditions prohibitive for any WT viral growth. These results demonstrate that defective hypermutated genomes can shape the phenotype of the circulating viral population. Partially active Vif alleles resulting in incomplete neutralization of cytoplasmic APOBEC3 molecules are directly responsible for the generation of a highly diverse, yet G-to-A biased, proviral reservoir, which can be exploited by HIV-1 to generate viable and drug-resistant progenies. PMID- 18391218 TI - Massive global ozone loss predicted following regional nuclear conflict. AB - We use a chemistry-climate model and new estimates of smoke produced by fires in contemporary cities to calculate the impact on stratospheric ozone of a regional nuclear war between developing nuclear states involving 100 Hiroshima-size bombs exploded in cities in the northern subtropics. We find column ozone losses in excess of 20% globally, 25-45% at midlatitudes, and 50-70% at northern high latitudes persisting for 5 years, with substantial losses continuing for 5 additional years. Column ozone amounts remain near or <220 Dobson units at all latitudes even after three years, constituting an extratropical "ozone hole." The resulting increases in UV radiation could impact the biota significantly, including serious consequences for human health. The primary cause for the dramatic and persistent ozone depletion is heating of the stratosphere by smoke, which strongly absorbs solar radiation. The smoke-laden air rises to the upper stratosphere, where removal mechanisms are slow, so that much of the stratosphere is ultimately heated by the localized smoke injections. Higher stratospheric temperatures accelerate catalytic reaction cycles, particularly those of odd nitrogen, which destroy ozone. In addition, the strong convection created by rising smoke plumes alters the stratospheric circulation, redistributing ozone and the sources of ozone-depleting gases, including N(2)O and chlorofluorocarbons. The ozone losses predicted here are significantly greater than previous "nuclear winter/UV spring" calculations, which did not adequately represent stratospheric plume rise. Our results point to previously unrecognized mechanisms for stratospheric ozone depletion. PMID- 18391219 TI - The conserved Wobble uridine tRNA thiolase Ctu1-Ctu2 is required to maintain genome integrity. AB - Modified nucleosides close to the anticodon are important for the proper decoding of mRNA by the ribosome. Particularly, the uridine at the first anticodon position (U34) of glutamate, lysine, and glutamine tRNAs is universally thiolated (S(2)U34), which is proposed to be crucial for both restriction of wobble in the corresponding split codon box and efficient codon-anticodon interaction. Here we show that the highly conserved complex Ctu1-Ctu2 (cytosolic thiouridylase) is responsible for the 2-thiolation of cytosolic tRNAs in the nematode and fission yeast. In both species, inactivation of the complex leads to loss of thiolation on tRNAs and to a thermosensitive decrease of viability associated with marked ploidy abnormalities and aberrant development. Increased level of the corresponding tRNAs suppresses the fission yeast defects, and our data suggest that these defects could result from both misreading and frame shifting during translation. Thus, a translation defect due to unmodified tRNAs results in severe genome instability. PMID- 18391220 TI - Complex interplays among population dynamics, environmental forcing, and exploitation in fisheries. AB - The patterns of variations in fisheries time series are known to result from a complex combination of species and fisheries dynamics all coupled with environmental forcing (including climate, trophic interactions, etc.). Disentangling the relative effects of these factors has been a major goal of fisheries science for both conceptual and management reasons. By examining the variability of 169 tuna and billfish time series of catch and catch per unit effort (CPUE) throughout the Atlantic as well as their linkage to the North Atlantic Oscillation (NAO), we find that the importance of these factors differed according to the spatial scale. At the scale of the entire Atlantic the patterns of variations are primarily spatially structured, whereas at a more regional scale the patterns of variations were primarily related to the fishing gear. Furthermore, the NAO appeared to also structure the patterns of variations of tuna time series, especially over the North Atlantic. We conclude that the patterns of variations in fisheries time series of tuna and billfish only poorly reflect the underlying dynamics of these fish populations; they appear to be shaped by several successive embedded processes, each interacting with each other. Our results emphasize the necessity for scientific data when investigating the population dynamics of large pelagic fishes, because CPUE fluctuations are not directly attributable to change in species' abundance. PMID- 18391221 TI - Tlx3 exerts context-dependent transcriptional regulation and promotes neuronal differentiation from embryonic stem cells. AB - The T cell leukemia 3 (Tlx3) gene has been implicated in specification of glutamatergic sensory neurons in the spinal cord. In cranial sensory ganglia, Tlx3 is highly expressed in differentiating neurons during early embryogenesis. To study a role of Tlx3 during neural differentiation, mouse embryonic stem (ES) cells were transfected with a Tlx3 expression vector. ES cells stably expressing Tlx3 were grown in the presence or absence of a neural induction medium. In undifferentiated ES cells, there was no significant difference in gene expression in the presence or absence of Tlx3, even after ES cells were cultured for an extensive time period. In contrast, expression levels of Mash1, Ngn1, and NeuroD were significantly higher in Tlx3-expressing cells after neural induction for 4 days compared with those in cells expressing the control vector. At 7 days after neural induction, whereas expression of the proneural genes was down-regulated, VGLUT2, GluR2, and GluR4 were significantly increased in ES cell-derived neurons expressing Tlx3. The sequential and coordinated expression of the proneural and neuronal subtype-specific genes identifies Tlx3 as a selector gene in ES cells undergoing neural differentiation. In addition, the differential effects of Tlx3 overexpression in undifferentiated ES cells compared with ES cell-derived neurons suggest that Tlx3 exerts context-dependent transcriptional signals on its downstream target genes. The context-dependent function of Tlx3 as a selector gene may be used to establish a novel strategy to conditionally generate excitatory glutamatergic neurons from ES cells to cure various types of neurodegenerative disorders. PMID- 18391222 TI - Presecretory oxidation, aggregation, and autophagic destruction of apoprotein-B: a pathway for late-stage quality control. AB - Hepatic secretion of apolipoprotein-B (apoB), the major protein of atherogenic lipoproteins, is regulated through posttranslational degradation. We reported a degradation pathway, post-ER pre secretory proteolysis (PERPP), that is increased by reactive oxygen species (ROS) generated within hepatocytes from dietary polyunsaturated fatty acids (PUFA). We now report the molecular processes by which PUFA-derived ROS regulate PERPP of apoB. ApoB exits the ER; undergoes limited oxidant-dependent aggregation; and then, upon exit from the Golgi, becomes extensively oxidized and converted into large aggregates. The aggregates slowly degrade by an autophagic process. None of the oxidized, aggregated material leaves cells, thereby preventing export of apoB-lipoproteins containing potentially toxic lipid peroxides. In summary, apoB secretory control via PERPP/autophagosomes is likely a key component of normal and pathologic regulation of plasma apoB levels, as well as a means for remarkably late-stage quality control of a secreted protein. PMID- 18391223 TI - Rare steroid receptor-negative basal-like tumorigenic cells in luminal subtype human breast cancer xenografts. AB - There are two major subtypes of human breast cancers: the luminal, estrogen, and progesterone receptor-positive, cytokeratin 18-positive (ER(+)PR(+)CK18(+)) subtype, and the basal ER(-)PR(-)CK18(-)CK5(+) subtype. Tumor-initiating cells (CD44(+)) have been described for human breast cancers; whether these are common to the two subtypes is unknown. We have identified a rare population of cells that are both CD44(+) and ER(-)PR(-)CK5(+) in luminal-like ER(+)PR(+) T47D human breast tumor xenografts. The tumor-isolated CD44(+) cell fraction was highly enriched for clonogenic (in vitro culture) and tumorigenic (in vivo reimplantation) cells compared with the CD44(-) cell fraction. Rare ER(-)PR( )CK5(+) cells were present within CD44(+)-derived colonies. Tumor-isolated cells placed in minimal media also contained rare ER(-)PR(-)CK5(+) cells at early time points (<10 cells); however, this population did not expand with increasing colony size. The number of ER(+)PR(+)CK5(-) cells, conversely, increased linearly with colony growth. Similary, tumors originating in vivo from CD44(+) cells contained a rare static ER(-)PR(-)CK5(+) population, an intermediate ER(-)PR( )CK5(-) population, and an expanding ER(+)PR(+)CK5(-) population. Putative ER(+)PR(+)CK5(+) transitional cells could be seen only in colonies or tumors treated with a progestin. We propose that luminal ER(+)PR(+) breast tumors contain a minor ER(-)PR(-)CK5(+) population that has the capacity to generate the majority of ER(+)PR(+)CK18(+)CK5(-) cells. Luminal breast cancers are treated with endocrine therapies that target ER. The rare ER(-)PR(-)CK5(+) progenitor cells would escape such treatments and survive to repopulate the tumor. PMID- 18391224 TI - Acute corticosterone treatment is sufficient to induce anxiety and amygdaloid dendritic hypertrophy. AB - Stress is known to induce dendritic hypertrophy in the basolateral amygdala (BLA) and to enhance anxiety. Stress also leads to secretion of glucocorticoids (GC), and the BLA has a high concentration of glucocorticoid receptors. This raises the possibility that stress-induced elevation in GC secretion might directly affect amygdaloid neurons. To address the possible effects of GC on neurons of amygdala and on anxiety, we used rats treated either acutely with a single dose or chronically with 10 daily doses of high physiological levels of corticosterone (the rat-specific glucocorticoid). Behavior and morphological changes in neurons of BLA were measured 12 days after the initiation of treatment in both groups. A single acute dose of corticosterone was sufficient to induce dendritic hypertrophy in the BLA and heightened anxiety, as measured on an elevated plus maze. Moreover, this form of dendritic hypertrophy after acute treatment was of a magnitude similar to that caused by chronic treatment. Thus, plasticity of BLA neurons is sufficiently sensitive so as to be saturated by a single day of stress. The effects of corticosterone were specific to anxiety, as neither acute nor chronic treatment caused any change in conditioned fear or in general locomotor activity in these animals. PMID- 18391226 TI - Lung compliance measurement in mice. PMID- 18391225 TI - DNA looping can enhance lysogenic CI transcription in phage lambda. AB - The lysogenic state of bacteriophage lambda is maintained by CI repressor, which negatively regulates two promoters to block lytic gene expression. Expression of CI is itself controlled by positive and negative feedback as CI binds to O(R) to regulate the P(RM) promoter. In addition to direct interactions with operator DNA, CI tetramers bound at O(L) and O(R) can come together to form an octamer, looping the DNA that lies between them and allowing O(L) to assist with negative regulation of P(RM). We used a fluorescent reporter protein to measure the CI concentration for a set of constructs that differ in their ability to assume various forms of the looped structure. Based on the observed steady-state fluorescence for these constructs, the presence of O(L) increases P(RM) activation unless both operators can be fully occupied. By calculating the probabilities for the underlying operator configurations present in each construct, two different models for the mechanism of enhanced activation allow us to predict that when the DNA is looped, P(RM) activation can be 2- to 4-fold higher than is possible for unlooped DNA. Based on our results, transcriptional regulation for lambda's lysogenic/lytic switch includes both activation and repression due to DNA looping. PMID- 18391229 TI - Positive Darwinian selection on crustacean hyperglycemic hormone (CHH) of the green shore crab, Carcinus maenas. AB - The tissue-specific expression and differential function of the crustacean hyperglycemic hormone (CHH) in Carcinus maenas indicate an interesting evolutionary history. Previous studies have shown that CHH from the sinus gland X organ (XO-type) has hyperglycemic activity, whereas the CHH from the pericardial organ (PO-type) neither shows hyperglycemic activity nor it inhibits Y-organ ecdysteroid synthesis. Here we examined the types of selective pressures operating on the variants of CHH in Carcinus maenas. Maximum likelihood-based codon substitution analyses revealed that the variants of this neuropeptide in C. maenas have been subjected to positive Darwinian selection indicating adaptive evolution and functional divergence among the CHH variants leading to two unique groups (PO and XO-type). Although the average ratio of nonsynonymous to synonymous substitution (omega) for the entire coding region is 0.5096, few codon sites showed significantly higher omega (10.95). Comparison of models that incorporate positive selection (omega > 1) with models not incorporating positive selection (omega <1) at certain codon sites failed to reject (p=0) evidence of positive Darwinian selection. PMID- 18391230 TI - Prediction of neurotoxins based on their function and source. AB - We have developed a method NTXpred for predicting neurotoxins and classifying them based on their function and origin. The dataset used in this study consists of 582 non-redundant, experimentally annotated neurotoxins obtained from Swiss Prot. A number of modules have been developed for predicting neurotoxins using residue composition based on feed-forwarded neural network (FNN), recurrent neural network (RNN), support vector machine (SVM) and achieved maximum accuracy of 84.19%, 92.75%, 97.72% respectively. In addition, SVM modules have been developed for classifying neurotoxins based on their source (e.g., eubacteria, cnidarians, molluscs, arthropods have been and chordate) using amino acid composition and dipeptide composition and achieved maximum overall accuracy of 78.94% and 88.07% respectively. The overall accuracy increased to 92.10%, when the evolutionary information obtained from PSI-BLAST was combined with SVM module of source classification. We have also developed SVM modules for classifying neurotoxins based on functions using amino acid, dipeptide composition and achieved overall accuracy of 83.11%, 91.10% respectively. The overall accuracy of function classification improved to 95.11%, when PSI-BLAST output was combined with SVM module. All the modules developed in this study were evaluated using five-fold cross-validation technique. The NTXpred is available at www.imtech.res.in/raghava/ntxpred/ and mirror site at http://bioinformatics.uams.edu/mirror/ntxpred. PMID- 18391231 TI - Genome-wide prediction and annotation of Burkholderia pseudomallei AraC/XylS family transcription regulator. AB - Many members of the AraC/XylS family transcription regulator have been proven to play a critical role in regulating bacterial virulence factors in response to environmental stress. By using the Hidden Markov Model (HMM) profile built from the alignment of a 99 amino acid conserved domain sequence of 273 AraC/XylS family transcription regulators, we detected a total of 45 AraC/XylS family transcription regulators in the genome of the Gram-negative pathogen, Burkholderia pseudomallei. Further in silico analysis of each detected AraC/XylS family transcription regulatory protein and its neighboring genes allowed us to make a first-order guess on the role of some of these transcription regulators in regulating important virulence factors such as those involved in three type III secretion systems and biosynthesis of pyochelin, exopolysaccharide (EPS) and phospholipase C. This paper has demonstrated an efficient and systematic genome wide scale prediction of the AraC/XylS family that can be applied to other protein families. PMID- 18391232 TI - BRM-Parser: a tool for comprehensive analysis of BLAST and RepeatMasker results. AB - BLAST and Repeat Masker Parser (BRM-Parser) is a service that provides users a unified platform for easy analysis of relatively large outputs of BLAST (Basic Local Alignment Search Tool) and RepeatMasker programs. BLAST Summary feature of BRM-Parser summarizes BLAST outputs, which can be filtered using user defined thresholds for hit length, percentage identity and E-value and can be sorted by query or subject coordinates and length of the hit. It also provides a tool that merges BLAST hits which satisfy user-defined criteria for hit length and gap between hits. The RepeatMasker Summary feature uses the RepeatMasker alignment as an input file and calculates the frequency and proportion of mutations in copies of repeat elements, as identified by the RepeatMasker. Both features can be run through a GUI or can be executed via command line using the standalone version. PMID- 18391233 TI - BTXpred: prediction of bacterial toxins. AB - This paper describes a method developed for predicting bacterial toxins from their amino acid sequences. All the modules, developed in this study, were trained and tested on a non-redundant dataset of 150 bacterial toxins that included 77 exotoxins and 73 endotoxins. Firstly, support vector machines (SVM) based modules were developed for predicting the bacterial toxins using amino acids and dipeptides composition and achieved an accuracy of 96.07% and 92.50%, respectively. Secondly, SVM based modules were developed for discriminating entotoxins and exotoxins, using amino acids and dipeptides composition and achieved an accuracy of 95.71% and 92.86%, respectively. In addition, modules have been developed for classifying the exotoxins (e.g. activate adenylate cyclase, activate guanylate cyclase, neurotoxins) using hidden Markov models (HMM), PSI-BLAST and a combination of the two and achieved overall accuracy of 95.75%, 97.87% and 100%, respectively. Based on the above study, a web server called 'BTXpred' has been developed, which is available at http://www.imtech.res.in/raghava/btxpred/. Supplementary information is available at http://www.imtech.res.in/raghava/btxpred/supplementary.html. PMID- 18391234 TI - In Silico analysis of voltage-gated sodium channel in relation to DDT resistance in vector mosquitoes. AB - The voltage-gated sodium channel (VGSC) is the target site for insecticides such as DDT and synthetic pyrethroids. A single base (A-T) change in the knock-down resistance (kdr) allele leads to an amino acid substitution at position 267 that confers the target-mediated resistance to DDT and synthetic pyrethroids in Anopheles gambiae. A theoretical model of the VGSC domain II that contains the site of mutation was constructed using the K;+ channel protein of Aeropyrum pernix as a template. The validated model with 88.6% residues in the favored region was subjected to the CASTp program that predicted 30 pockets in the modeled domain II for ligand interaction. In the model, at position 267, leucine was manually replaced with phenylalanine. When this altered model was subjected to the CASTp program, the search results showed the same number of pockets. The docking results indicate that DDT interacts with the modeled VGSC domain II at position 275 in the presence of leucine or in the presence of phenylalanine (binding energy =-5.32 kcal/mol, -6.21 kcal/mol). It appears from the results that the mutation at position 267 has no direct influence on the interaction of DDT with the target protein. Therefore, to understand the interaction affinity of DDT with the target and influence of the mutation on the existence of active sites/pockets in relation to ligand binding, a whole VGSC model is necessary. PMID- 18391235 TI - Evaluation of codon bias perspectives in phage therapy of Mycobacterium tuberculosis by multivariate analysis. AB - To reveal the relative synonymous codon usage and base composition variation in bacteriophages, six mycobacteriophages were used as a model system here and both parameters in these phages and their host bacteria, Mycobacterium tuberculosis, have been determined and compared. As expected for GC-rich genomes, there are predominantly G and C ending codons in all 6 phages. Both N_{c} plot and correspondence analysis on relative synonymous codon usage indicate that mutation bias and translation selection influences codon usage variation in the 6 phages. Further analysis indicates that among 6 Mycobacterium phages Che9c, Bxz1 and TM4 may be extremely virulent in nature as most of their genes have high translation efficiency. Based on our data we suggest that the genes of above three phages are expressed rapidly by host's translation machinery. The information might be used to select the extremely virulent Mycobacterium tuberculosis phages suitable for phage therapy. PMID- 18391236 TI - Clustering gene expression data using graph separators. AB - Recent work has used graphs to modelize expression data from microarray experiments, in view of partitioning the genes into clusters. In this paper, we introduce the use of a decomposition by clique separators. Our aim is to improve the classical clustering methods in two ways: first we want to allow an overlap between clusters, as this seems biologically sound, and second we want to be guided by the structure of the graph to define the number of clusters. We test this approach with a well-known yeast database (Saccharomyces cerevisiae). Our results are good, as the expression profiles of the clusters we find are very coherent. Moreover, we are able to organize into another graph the clusters we find, and order them in a fashion which turns out to respect the chronological order defined by the the sporulation process. PMID- 18391238 TI - Clustering formal concepts to discover biologically relevant knowledge from gene expression data. AB - The production of high-throughput gene expression data has generated a crucial need for bioinformatics tools to generate biologically interesting hypotheses. Whereas many tools are available for extracting global patterns, less attention has been focused on local pattern discovery. We propose here an original way to discover knowledge from gene expression data by means of the so-called formal concepts which hold in derived Boolean gene expression datasets. We first encoded the over-expression properties of genes in human cells using human SAGE data. It has given rise to a Boolean matrix from which we extracted the complete collection of formal concepts, i.e., all the largest sets of over-expressed genes associated to a largest set of biological situations in which their over expression is observed. Complete collections of such patterns tend to be huge. Since their interpretation is a time-consuming task, we propose a new method to rapidly visualize clusters of formal concepts. This designates a reasonable number of Quasi-Synexpression-Groups (QSGs) for further analysis. The interest of our approach is illustrated using human SAGE data and interpreting one of the extracted QSGs. The assessment of its biological relevancy leads to the formulation of both previously proposed and new biological hypotheses. PMID- 18391237 TI - Differential genome analyses of metabolic enzymes in Pseudomonas aeruginosa for drug target identification. AB - Complete genome sequences of several pathogenic bacteria have been determined, and many more such projects are currently under way. While these data potentially contain all the determinants of host-pathogen interactions and possible drug targets, computational tools for selecting suitable candidates for further experimental analyses are currently limited. Detection of bacterial genes that are non-homologous to human genes, and are essential for the survival of the pathogen represents a promising means of identifying novel drug targets. We used a differential pathway analyses approach (based on KEGG data) to identify essential genes from Pseudomonas aeruginosa. Our approach identified 214 unique enzymes in P. aeruginosa that may be potential drug targets and can be considered for rational drug design. About 40% of these putative targets have been reported as essential by transposon mutagenesis data elsewhere. Homology model for one of the proteins (LpxC) is presented as a case study and can be explored for in silico docking with suitable inhibitors. This approach is a step towards facilitating the search for new antibiotics. PMID- 18391239 TI - In Silico identification and characterization of mRNA-like noncoding transcripts in Medicago truncatula. AB - Accumulating evidence suggests that that non-coding RNAs (ncRNAs) play key roles in gene regulation and may form the basis of an inter-gene communication system. Many ncRNAs are synthesized similar to mRNAs and can be detected through screening of polyA-rich EST or cDNA libraries. We developed a computational pipeline to screen EST and genomic sequence data for those transcribed genes with limited protein coding potential and applied this pipeline to the model legume Medicago truncatula. This process identified a set of 503 mRNA-like transcripts that appear not to encode proteins. Further computational analysis showed that many of these ncRNA candidates share structural similarities to known ncRNAs and that they clearly differ from protein coding genes and non-transcribed regions in their base and oligonucleotide compositions, as well as in aspects of secondary structure. By using a machine learning approach, we show that the distinctive ncRNA features presented in this study can be used to discriminate most ncRNAs and may thus be useful for improving ncRNA prediction. Computational analysis of EST isolation frequencies in various plant tissues showed that the expression levels and expression profiles of the putative ncRNAs and mRNAs differ - most interestingly, the putative ncRNAs are highly expressed relative to mRNAs in the root nodule tissue and conserved only in closely related plants. The work presented here constitutes the first large-scale prediction and characterization of ncRNAs in legumes, and provides a basis for further research on elucidating ncRNA function in legume genomics. PMID- 18391240 TI - The CAI Analyser Package: inferring gene expressivity from raw genomic data. AB - The Codon Adaptation Index (CAI) was introduced by Sharp and Li in 1987 to quantify codon usage similarities between a coding sequence and a set of reference sequences. When synonymous codons for a given amino acid exist, highly expressed genes seem to prefer some of them, according to tRNA abundance and thermodynamic issues. Some authors have described CAI-based methods to derive expressivity measures for all genes in a genome, in a computational framework. Here we present the CAIAP (CAI Analyser Package), a platform independent package of computer programs allowing the calculation of the CAI and a deep study of gene expressivity from raw gene sequences. Our approach implements and optimizes a procedure to derive the reference sequences from whole genomes and use their codon usage for CAI estimation. Moreover, a set of analysis tools are provided to perform statistical analyses and therefore to give robustness to results. OBJECTIVE: Our efforts were aimed to produce an easy-to-use and fully automatic set of programs specifically designed for the analysis of gene expressivity and inter-species comparisons on a great number of genomes. Moreover, the output integrates information coming from functional annotations of genes. We are maintaining a web server storing our analyses for hundreds of genomes, allowing intergenomic comparison of data thanks to dedicated search engines. The CAIAP server is hosted at www4.unifi.it/scibio/bioinfo/caiap/html. The programs (maintained as Perl scripts) are also available for download at the same location. PMID- 18391241 TI - Correlation between gene silencing activity and structural features of antisense oligodeoxynucleotides and target RNA. AB - Antisense oligonucleotides inactivate mRNA targets, providing a tool for post transcriptional gene silencing and a potential novel treatment for many diseases. Reliable design of active antisense depends on better understanding of the mechanism of antisense-target RNA interaction. We have studied the correlation between activity of antisense oligodeoxynucleotides (ASO) and structural features of both antisense and target RNAs. A total of 348 ASOs with known activities and their target RNA sequences are classified into categories according to their predicted secondary structural features. Statistical analysis showed that higher activity is more likely to happen at RNA stem-loops than at other RNA structural categories. The data suggest a weak correlation between the stability of ASO structure and activity. Remarkably, a structural fit between ASO and target seems important for antisense activity. Significantly higher antisense activity is achieved with stem-loop ASOs on stem-loop or linear RNA targets. PMID- 18391242 TI - BETTY: prediction of beta-strand type from sequence. AB - Most secondary structure prediction programs do not distinguish between parallel and antiparallel beta-sheets. However, such knowledge would constrain the available topologies of a protein significantly, and therefore aid existing fold recognition algorithms. For this reason, we propose a technique which, in combination with existing secondary structure programs such as PSIPRED, allows one to distinguish between parallel and antiparallel beta-sheets. We propose the use of a support vector machine (SVM) procedure, BETTY, to predict parallel and antiparallel sheets from sequence. We found that there is a strong signal difference in the sequence profiles which SVMs can efficiently extract. With strand type assignment accuracies of 90.7% and 83.3% for antiparallel and parallel strands, respectively, our method adds considerably to existing information on current 3-class secondary structure predictions. BETTY has been implemented as an online service which academic researchers can access from our website http://www.fz-juelich.de/nic/cbb/service/service.php. PMID- 18391243 TI - PGV - Prokaryotic Genome Viewer. AB - Recent sequencing of genomes of several microorganisms provides an opportunity to have access to huge volumes of data stored in various databases. This has resulted in the development of various computational and visualization tools to aid in retrieval and analysis of data. Development of user friendly genome data mapping and visualization tools facilitates researchers to closely examine various features of genes and make inferences from the displayed data efficiently. PGV - Prokaryotic Genome Viewer is a Java based web application tool capable of generating high quality interactive circular chromosome maps. With simple mouse roll over tasks on the interested region on the displayed map, the user is provided with features such as feature labeling, multi-fold zooming, image rotation and hyperlinking to different information resources. The tool is capable of instantaneously generating maps using user-supplied sequence data. PMID- 18391244 TI - Analysis of codon usage patterns and predicted highly expressed genes for six phytopathogenic Xanthomonas genomes shows a high degree of conservation. AB - Members of the genus Xanthomonas are significant phytopathogens, which cause diseases in several economically important crops including rice, canola, tomato, citrus, etc. We have analyzed the genomes of six recently sequenced Xanthomonas strains for their synonymous codon usage patterns for all of protein coding genes and specific genes associated with pathogenesis, and determined the predicted highly expressed (PHX) genes by the use of the codon adaptation index (CAI). Our results show considerable heterogeneity among the genes of these moderately G+C rich genomes. Most of the genes were moderate to highly biased in their codon usage. However, unlike ribosomal protein genes, which were governed by translational selection, those genes associated with pathogenesis (GAP) were affected by mutational pressure and were predicted to have moderate to low expression levels. Only two out of 339 GAP genes were in the PHX category. PHX genes present in clusters of orthologous groups of proteins (COGs) were identified. Genes in the plasmids present in two strains showed moderate to low expression level and only a couple of genes featured in the PHX list. Common genes present in the top-20 PHX gene-list were identified and their possible functions are discussed. Correspondence analysis showed that genes are highly confined to a core in the plot. PMID- 18391245 TI - Collaborative voxel-based surgical virtual environments. AB - Virtual Reality-based surgical simulators can utilize Collaborative Virtual Environments (C-VEs) to provide team-based training. To support real-time interactions, C-VEs are typically replicated on each user's local computer and a synchronization method helps keep all local copies consistent. This approach does not work well for voxel-based C-VEs since large and frequent volumetric updates make synchronization difficult. This paper describes a method that allows multiple users to interact within a voxel-based C-VE for a craniotomy simulator being developed. Our C-VE method requires smaller update sizes and provides faster synchronization update rates than volumetric-based methods. Additionally, we address network bandwidth/latency issues to simulate networked haptic and bone drilling tool interactions with a voxel-based skull C-VE. PMID- 18391246 TI - Eye gaze tracking for endoscopic camera positioning: an application of a hardware/software interface developed to automate Aesop. AB - A redesigned motion control system for the medical robot Aesop allows automating and programming its movements. An IR eye tracking system has been integrated with this control interface to implement an intelligent, autonomous eye gaze-based laparoscopic positioning system. A laparoscopic camera held by Aesop can be moved based on the data from the eye tracking interface to keep the user's gaze point region at the center of a video feedback monitor. This system setup provides autonomous camera control that works around the surgeon, providing an optimal robotic camera platform. PMID- 18391247 TI - Repeated palpatory training of medical students on the Virtual Haptic Back. AB - The effectiveness of simulation-based training has been accepted with great success in many fields including medicine. Most of the simulation research and development in medicine has focused on surgery. There has been some development of hardware based biomechanical models of sections of human anatomy, such as pelvic exam simulators. More recently, with the advances in haptics technology, software and hardware based simulators are being developed for the previously ignored area of palpatory diagnosis. The Virtual Haptic Back (VHB) is a simulator based on virtual reality and haptics that is currently being used to train medical students in palpatory diagnosis. This study examined the effect of repeating the training on the VHB. PMID- 18391248 TI - Strategies to reduce patient harm: understanding the role of design and the built environment. AB - One of the great ironies of modern medicine is that the very environments created to heal are the cause of countless injuries, illnesses and death to the vulnerable population they were created to serve. Not since Florence Nightingale visited the pestholes in which wounded and sick British soldiers were housed in the Crimea, has there been a growing international awareness of the harm caused by the healing environment. There is growing recognition that risks and hazards of health care associated injury and harm are a result of systemic design problems rather than poor performance by individual providers. The evidence is overwhelming that current hospital design is not sufficient to prevent medical errors, rates of infection and injuries from falls, and even contributes to slow patient recovery and high nurse turnover.Today's challenges are even steeper than a half-century ago due to: a) Rising concern and demand about health care costs and quality, b) Large population increase with an exponentially growing elderly proportion, c) A proliferation of complex and confusing local, state and federal regulations, d) A dramatically evolving technological environment, e) A sicker, more debilitated and compromised patient population, and f) A workforce with higher expectations of the work environment should be like. The present hospital construction boom provides an opportunity to rethink hospital design, and especially to consider how improved hospital design can help reduce staff stress and fatigue, reduce patient and family stress and improve patient outcomes and quality of care. A key challenge remains how to incorporate technology into the local learning environment and culture in ways that optimize its implementation and use. PMID- 18391250 TI - Learning & retention in adaptive serious games. AB - Serious games are being actively explored as supplements to and, in some cases, replacement for traditional didactic lectures and computer-based instruction in venues ranging from medicine to the military. As part of an intelligent tutoring system (ITS) for nuclear event first responders, we designed and evaluated two serious games that were integrated with adaptive multimedia content. Results reveal that there was no decay in score six weeks following game-based training, which contrasts with results expected with traditional training. This study suggests that adaptive serious games may help integrate didactic content presented though conventional means. PMID- 18391249 TI - A non-linear mass-spring model for more realistic and efficient simulation of soft tissues surgery. AB - An extension to the classical mass-spring model for more realistic simulation of soft tissues for surgery simulation was proposed. The conventional equations of mass-spring model were generalized for non-linear springs, and model parameters were tuned using experimental data. Results show that the proposed model is fast and interactive, and also demonstrate the typical nonlinear and visco-elastic behaviors of soft tissues well. PMID- 18391251 TI - Direct versus indirect laryngoscopic visualization in human endotracheal intubation: a tool for virtual anesthesia practice and teleanesthesiology. AB - When performing the ABC's of care for the trauma patient, airway management is of paramount importance. Management of the airway is often difficult because medical personnel caring for the patient do not commonly intubate patients or manage airways. To accomplish endotracheal intubation, a direct line of sight must be accomplished through the mouth, pharynx and larynx to the glottic opening. This is anatomically challenging in patients with a small mouth, large tongue, lack of cervical mobility, cervical trauma, protruding incisors or small mandible. This investigation compares indirect laryngoscopy, which allows the laryngoscopist to "see around the corner" during intubation, to standard direct laryngoscopy. This indirect view is a virtual view of the airway accomplished by projecting the view from an image device on the end of the laryngoscope to a monitor viewed by the laryngoscopist. The virtual (indirect) laryngoscopy improved the view of the glottic opening by an average 1.28 (p<0.001) Cormack-Lehane grades, consistant with existing literature. Indirect laryngoscopy results in improved glottic visualization compared to direct laryngoscopy. This difference will prove critically important for medical personnel who infrequently intubate and for students learning intubation skills in a clinical setting. The results of our study confirm the value of videolaryngoscopy as a standard method for hands-on airway management training of medical personnel. Indirect "virtual" laryngoscopy is an advanced technology method which will advance the development of teleanesthesiology practice. Videolaryngoscopy is an enabling technology for development of remote telementoring of trainee intubation skills curricula using video enabled distributed learning systems. This research was conducted following an approved University of Nebraska Medical Center institutional review board protocol. PMID- 18391252 TI - Far forward feasibility: testing a cricothyroidotmy simulator in Iraq. AB - This study describes our experience with using a virtual reality simulator, CricSim, to enhance the training of combat medics to perform a cricothyroidotomy (surgical airway) while in Iraq. Over a six month period, 65 medics used the simulator as part of a Combat Medic Advanced Skills Training class while in Iraq and were asked to evaluate it. Students self-assessed comfort level with the procedure improved dramatically from baseline (p<5.6 x 10(-17)). The CricSim was rated highly on realism but only moderately on ease of use. The use of this simulator in a far forward setting was feasible, enhanced training, and provided necessary end-user feedback for future development of this training platform. PMID- 18391253 TI - Evaluating the clinical utility of stereoscopic clinical photography. AB - Stereoscopic imaging during clinical evaluation can provide utility to physicians and medical educators by extending clinical photography with the provision of depth. A digital SLR camera body fitted with a stereoscopic lens and an autostereoscopic display are evaluated. This paper describes the image acquisition workflow and post-processing methods required to incorporate stereoscopic images into an electronic medical record for physician review. PMID- 18391255 TI - Trocar insertion: the neglected task of VR simulation. AB - We hypothesized that simulation based practice will results in skill improvement on trocar insertion task. Novices, junior trainees and surgeons inserted a trocar into a simulated abdomen. The depth of penetration (plunge) was the similar for novice and junior trainees (p=.98), and exceeded that observed for practicing surgeons (p<.05). Trocar insertion may be an important variable to consider for surgical skill training. To date, no virtual reality (VR) trainers allow for practice of this skill. We will present a prototype of a VR trainer designed. PMID- 18391254 TI - Validating advanced robot-assisted laparoscopic training task in virtual reality. AB - The purpose of this study was to validate a complex robotic surgical task, mesh alignment, in virtual reality. Nine subjects unrolled and aligned a mesh onto an inanimate template for the mesh alignment task in both an actual (the da Vinci Robotic Surgical System) and a virtual environment. Data analysis included time to task completion, distance traveled, and speed, of the surgical instrument, as well as electromyography of the extensors and flexors of the dominant arm of the subject. Paired t-tests were used to compare the dependent variables between the actual and virtual environments. The virtual mesh alignment task was statistically similar for all variables except the flexor activity as compared to the actual task. In conclusion, virtual reality could be used as an effective environment to train the next generation of robot-assisted laparoscopic surgeons. PMID- 18391256 TI - Efficient soft tissue deformation using charged particles. AB - As the power of personal computing increases, there is a greater demand for high fidelity virtual environments for medical training simulators. However, it is still a challenge to provide realistic soft tissue deformation especially when haptic feedback is also required. This paper presents a new approach to soft tissue deformation using a novel Charged Particle method to efficiently model both the structure and haptic properties of anatomy in real time. PMID- 18391257 TI - Simulation of dynamic ultrasound based on CT models for medical education. AB - In our approach, we first specify a 3D model of the image structure by segmenting a CT dataset into the respective tissues followed by assigning the acoustic properties (velocity, impedance, scattering mean and standard deviation, damping factor and packing factor). Given that model, we simulate the ray propagation, beam forming, and finally the backscattering. Due to the inhomogenities of tissue, different physical models for ultrasound simulation are required: Rayleigh scattering is applied for homogenous regions and ray tracing techniques handle abrupt changes in acoustic impedance on tissue boundaries. The latter leads to different phenomena like refraction (Snell's law), reflection and transmission (Fresnel equation). The gradients needed for these methods are precomputed for each model using a central-difference method with multiple neighbours. Absorption is calculated by the Beer-Lambert law. PMID- 18391258 TI - Steps towards open standards for medical virtual reality systems. AB - Very little medical virtual reality systems which are developed are applied in real surgical scenarios. One reason for this is that the system solutions resulting from research projects often address a single research question and are not embedded in an overall design. This paper presents a DICOM based approach on standardizing data structures, i.e. surface meshes, which are required for supporting surgical workflows by virtual reality applications. PMID- 18391259 TI - Patient specific physical anatomy models. AB - The advent of small footprint stereo-lithographic printers and the ready availability of segmentation and surface modeling software provide a unique opportunity to create patient-specific physical models of anatomy, validation of image guided intervention applications against phantoms that exhibit naturally occurring anatomic variation. Because these models can incorporate all structures relevant to a procedure, this allows validation to occur under realistic conditions using the same or similar techniques as would be used in a clinical application. This in turn reduces the number of trials and time spent performing in-vivo validation experiments. In this paper, we describe our general approach for the creation of both non-tissue and tissue-mimicking patient-specific models as part of a general-purpose patient emulation system used to validate image guided intervention applications. PMID- 18391260 TI - Towards developing an immersive and interactive virtual fairview peri-operative environment. AB - We discuss the goal and a design of an immersive and interactive virtual fairview peri-operative environment for patient education for surgery. We further discuss a pipeline for building 3D models of peri-operative environment for usage in VR. Specifically, we capitalize on the recent advancement on laser scanning technology to perform such modeling for both efficiency and accuracy. PMID- 18391261 TI - Towards anatomical modelling of multiple organs interaction using real time GPU based non-linear elasticity. AB - Finite element methods (FEM) have been used extensively within the field of surgical simulation to describe physically realistic interactions with soft tissue organs. However, FEMs require researchers to balance computational costs against approximation techniques to maintain adequate performance. We aim to extend previous work in nonlinear FEM implementation with extensions which improve the performance of accurate modelling of multiple organs undergoing deformations. PMID- 18391262 TI - Visualizing the medical modeling and simulation database: trends in the research literature. AB - Medical modeling and simulation literature has undergone significant changes from 2000 until 2007. The MMVR Conference and the MMSD provide two avenues for observing trends in medical modeling and simulation literature. The literature submitted to MMVR and MMSD from 2000 to 2007 were placed into 8 categories and assessed through meta-analysis. PMID- 18391263 TI - Effects of repeated CPR training in virtual worlds on medical students' performance. AB - We report on a study that investigates the relationship between repeated training of teams managing a medical emergency (CPR) in a Virtual World and performance outcome measures in a group of 12 medical students. The focus of the training was on individual actions, but also on interaction and behavior in the team. Current CPR training seems to lack important team training aspects which this type of training is addressing. Although a pilot study, we found clear indications of improved performance related to reduced number of errors and an increased CPR efficiency. This type of educational technology could be expanded to other groups for a similar purpose because of its easiness to use, adaptability and interactivity. PMID- 18391264 TI - A flexible, conformal ultrasound array for medical imaging. AB - An ultrasound imaging system is under development that features a flexible transducer that can be wrapped conformally around curved surfaces of the body. The device is intended to deliver high image quality without the need for mechanical scanning, and will benefit medical personnel with limited ultrasound training. A 2 x 8 element piezoelectric transducer prototype and 15.5 MHz imaging system have been built, and the system concept has been demonstrated by imaging a soft tissue phantom. The transducer design, imaging procedure, and preliminary image processing techniques are presented. PMID- 18391265 TI - Novel virtual Lap-Band simulator could promote patient safety. AB - This paper presents, for the first time, a physics-based modeling technique for the Lap-Band (Inamed Health) used in laparoscopic gastric banding (LAGB) operations for treating the morbidly obese. A virtual LAGB simulator can help train medical students as well as surgeons who embark at learning this relatively new operation. The Lap-Band has different thickness and curvature along the centerline, and therefore leads to different deformation behaviors. A hybrid modeling strategy is therefore adopted to successfully replicate its dynamics. A mass-spring model, used to model the less stiff part, is coupled to a quasi static articulated link model for the more stiff and inextensible part. The virtual Lap-Band model has been implemented into a complete graphics-haptics physics-based system with two PHANToM Omni devices (from Sensible Technologies) being used for real-time bimanual interaction with force feedback. PMID- 18391266 TI - Medical student satisfaction using a virtual patient system to learn history taking communication skills. AB - Virtual patients (VPs) have the potential to augment existing medical school curricula to teach history-taking and communication skills. A goal of our current efforts to study virtual characters in health professions education is to develop a system that can be independently accessed and thus user satisfaction is an important factor in how readily this technology will be adopted. Twenty-three medical students participated in a study in which they interviewed a virtual patient and were asked to rate the educational value of the experience. Despite some of the limitations in this developing technology, students were generally receptive to its use as an educational tool. Further enhancements to the system, including increased fidelity of the interaction and novel feedback mechanisms, should improve learner satisfaction with and adoption of the virtual patient system. PMID- 18391267 TI - A simulator to explore the role of haptic feedback in cataract surgery training. AB - Phacoemulsification cataract surgery, a minimally invasive technique to remove a cloudy lens from the eye, is one of the most commonly performed surgical procedures in the western world. Conventional training for this procedure involves didactic lectures and practice on pig and human cadaver eyes, none of which allow trainees to form an accurate predictive model of human tissue behavior during surgery. A virtual environment simulator for capsulorrhexis, one of the first steps in cataract surgery, has been developed that allows a trainee to use surgical instruments to excise a circle of tissue on the anterior side of the lens capsule through tearing. The simulator invokes a deformable mass-spring damper mesh model of the tissue that can be grasped and torn via shearing. A novel algorithm for mesh division and maintenance enables realistic tearing behavior. The trainee controls tool motion using a 3-degree-of-freedom haptic device, and haptic feedback is provided from the virtual tissue. Although the haptic feedback in a real capsulorrhexis procedure is below the human threshold of haptic sensing, this simulator enables an experiment to determine the effectiveness of "haptic training wheels" -- the idea of haptic training for a task without haptic feedback. PMID- 18391268 TI - Training and evaluation of lumbar punctures in a VR-environment using a 6DOF haptic device. AB - A virtual reality system for the training of the lumbar puncture intervention is presented. We use a haptic device with six degrees of freedom (6DOF) to feedback forces that resist needle insertion and rotation. An improved haptic volume rendering approach is used to calculate the forces and an evaluation component has been developed to rate the success of virtual lumbar punctures to trace the training process of the user and to give feedback about failures. PMID- 18391269 TI - A prototype haptic feedback system for lower-limb prostheses and sensory neuropathy. AB - Lower-limb sensory loss as a result of peripheral neuropathy or amputation results in sub-optimal movement and an increased incidence of injury. While the adoption of lower-limb prostheses and therapeutic footwear can reduce tissue injury and support movement, the fundamental problem of sensory loss continues to exist. A prototype haptic feedback system has been developed to aid in the recovery of lower-limb sensation due to these causes. Thin-film force sensors placed at the critical points for gait and balance functions collect essential force data, which is delivered to the user via pneumatically controlled balloon inflation. It is postulated that the use of this system will increase the tactile awareness of a patient's lower-limb or prosthesis, and when used in concert with modern rehabilitation techniques will create a method that will reduce the duration and improve the quality of lower-limb rehabilitation, especially in gait and balance functions. PMID- 18391270 TI - A framework for interactive examination of automatic segmented tumors in a virtual environment. AB - An immersive virtual environment for viewing and interacting with three dimensional representations of medical image data is presented. Using a newly developed automatic segmentation method, a segmented object (e.g., tumor or organ) can also be viewed in the context of the original patient data. Real time interaction is established using joystick movements and button presses on a wireless gamepad. Several open-source platforms have been utilized, such as DCMTK for processing of DICOM formatted data, Coin3D for scenegraph management, SimVoleon for volume rendering, and VRJuggler to handle the immersive visualization. The application allows the user to manipulate representations with features such as fast pseudo-coloring to highlight details of the patient data, windowing to select a range of tissue densities for display, and multiple clipping planes to allow the user to slice into the patient. PMID- 18391271 TI - Interactive visuo-haptic surgical planning tool for pelvic and acetabular fractures. AB - Treatment of pelvic and acetabular fractures still poses a major challenge to trauma surgeons. We present a tool for intervention planning for such injuries using patient-specific models built from Computed Tomography data. The presented tool has three main parts: (1) the virtual reduction of the bone fragments, (2) the virtual adaptation of the osteosynthesis implants and (3) Finite Element Analysis (FEA) for testing mechanical behavior of the resulting intervention plan. Our tool provides an intuitive visuo-hapic interface designed to be used by trauma surgeons. The type and size of the osteosynthesis material can be determined and measurements like distances and angles relative to landmarks can be taken. First results of prospectively planned interventions show an excellent correlation and a significant gain in operation time. PMID- 18391272 TI - Optimizing a head-tracked stereo display system to guide hepatic tumor ablation. AB - Radio frequency ablation is a minimally invasive intervention that introduces -- under 2D ultrasound guidance and via a needle-like probe -- high-frequency electrical current into non-resectable hepatic tumors. These recur mostly on the periphery, indicating errors in probe placement. Hypothesizing that a contextually correct 3D display will aid targeting and decrease recurrence, we have developed a prototype guidance system based on a head-tracked 3D display and motion-tracked instruments. We describe our reasoning and our experience in selecting components for, designing and constructing the 3D display. Initial candidates were an augmented reality see-through head-mounted display and a virtual reality "fish tank" system. We describe the system requirements and explain how we arrived at the final decision. We show the operational guidance system in use on phantoms and animals. PMID- 18391273 TI - Why do you drink? Virtual reality as an experiential medium for the assessment of alcohol-dependent individuals. AB - The aim of this study is to assess social, personality and behaviors of alcoholics using Virtual Reality (VR). Specifically, we defined a VR protocol using the free NeuroVR software -- (http://www.neurovr.org) -- to investigate the following factors: drinking behaviors, intrapersonal factors (Emotional Management and Self Esteem) and environmental factors (Relational Competences and Social Pressure). In this preliminary study we evaluated the difference between assessment methods by comparing the VR assessment protocol with the SCID - Structured Clinical Interview for DSM-IV Axis I Disorders - in a sample of 20 alcohol-dependent individuals (10 experimental group + 10 control group) entering a non-pharmacological outpatient treatment. The data, obtained using both qualitative and quantitative analyses, confirm the possibility of using the VR protocol in the assessment of alcohol-dependent patients: the therapist obtains more critical data about behaviors and attitudes in less time. Further, the VR group reported a significant improvement in the motivation for change after the assessment protocol, not found in the SCID group: apparently, the experiential approach required by VR makes the patient more active and involved in the processes of introspection and change. A wider sample and a multicentric trial are now needed to confirm these results. PMID- 18391274 TI - Low cost augmented reality for training of MRI-guided needle biopsy of the spine. AB - In needle biopsy of the spine, an Augmented Reality (AR) image guidance system can be very effective in ensuring that while targeting the lesion with the biopsy needle, vital organs near the spine are not damaged and that the approach path is accurate. This procedure requires skill that is hard to master on patients. In this paper, we present a low cost AR based training set-up which consists of a software that uses one static single-camera tracking mechanism to locate the biopsy needle in the patient and which then augments the camera feed of the patient with virtual data providing real-time guidance to the surgeon for insertion of the biopsy needle. The setup is implemented using a phantom model consisting of a set of carefully modeled holes to simulate the needle insertion task. The lack of requirement of elaborate infrared tracking systems and high computing power makes this system very effective for educational and training purposes. PMID- 18391275 TI - The instrumented instrument: characterization and training of manual skill in open suturing. AB - Instruction in the use of forceps and needle driver to suture tissues typically involves group didactic and demonstration sessions, followed by limited individual observation and coaching. Most motor learning required for dexterous suturing takes place during unsupervised practice with practice boards or during actual procedures in the operating room. We are developing surgical instruments with embedded microelectromechanical sensors for tracking instrument motion. Motion data is acquired and processed on a computer for concurrent or summary performance feedback during practice. An integral feature in our approach is the use of digital video recordings, synchronized with the sensor signals, to parse surgical procedures into a series of actions based on a task analysis. We envision the parsed video and signals as tools for assessment and performance feedback that an instructor could use to offer more extended individualized coaching. In a small pilot study, we concentrated our data analysis on the orientation of a needle driver about its long axis, the range of motion in one throw, and the timing of subtasks. PMID- 18391276 TI - Ergonomic usability testing of operating room devices. AB - Medical errors in the operating room due to design, technological factors and poor human factors engineering are common. Determining the factors which facilitate or hinder successful performance during the use of medical systems is critical to the creation of standardized metrics for usability of medical devices in the operating room. Creation of a standardized evaluation for usability based on international standards and user-centered design principles can help determine what role these factors have in medical errors. In addition, device design can be evaluated to promote safety, measured objectively by metrics such as reduction in medical errors with a decrease in performance time. PMID- 18391277 TI - Towards an in vivo wireless mobile robot for surgical assistance. AB - The use of miniature in vivo robots that fit entirely inside the peritoneal cavity represents a novel approach to laparoscopic surgery. Previous work has demonstrated that mobile and fixed-base in vivo robots can be used to improve visualization of the surgical field and perform surgical tasks such as collecting biopsy tissue samples. All of these robots used tethers to provide for power and data transmission. This paper describes recent work focused on developing a modular wireless mobile platform that could be used for in vivo robotic sensing and manipulation applications. One vision for these types of self-contained in vivo robotic devices is that they could be easily carried and deployed by non medical personnel at the site of an injury. Such wireless in vivo robots are much more transportable and lower cost than current robotic surgical assistants, and could ultimately allow a surgeon to become a remote first responder irrespective of the location of the patient. PMID- 18391278 TI - Design and implementation of rule-based medical models: an In Silico patho physiological trauma model for hypovolemic shock. AB - Trauma from 'Dirty' Bomb blasts presents complex clinical problems to healthcare providers who must make critical emergency care decisions with incomplete information, usually limited initially only to cursory observations and vital signs. A set of simple, HFSM patho-physiological models of hypovolemic shock based upon blood volume deficits and remedial therapeutic actions has been created for 10 Virtual World scenarios used for training healthcare personnel in the diagnosis and management of 'dirty' bomb victims. Several general rules define the models: * Virtual patients have individual characteristics of gender, age, health status. * An Expanded Organ Injury Scale (EOIS) indicates in descriptive terms the type, severity, and location of the injury. * Traumatic Hemorrhage Classes (THC) determine Blood Volume Deficit. * EOIS/THC Score and Volume Deficit are tightly linked to the Hemorrhage Rate. * Remedial actions include Wound care, Volume Replacement (0.9% saline, packed cells), and a Drug Formulary for Treating Hemorrhagic Shock. The above rules and their logical basis are derived from the literature on emergency medicine (trauma), with modifications for organ injury, and introducing progressive deterioration beyond the body's homeostatic capabilities, into failure of systems and organs, followed by death. The ten unique models and scenarios are designed to run simultaneously in a virtual emergency department to provide training experiences for healthcare personnel with trauma from a 'dirty' bomb blast event. PMID- 18391279 TI - A modular simulation framework for colonoscopy using a new haptic device. AB - We have developed a multi-threaded framework for colonoscopy simulation utilising OpenGL with an interface to a real-time prototype colonoscopy haptic device. A modular framework has enabled us to support multiple haptic devices and efficiently integrate new research into physically based modelling of the colonoscope, colon and surrounding organs. The framework supports GPU accelerated algorithms as runtime modules, allowing the real-time calculations required for haptic feedback. PMID- 18391280 TI - First clinical evaluation of the navigated controlled drill at the lateral skull base. AB - Surgery on the lateral skull base puts delicate structures at risk. To support the surgeon in identifying and protecting the risk structures the principle of Navigated Control (NC) can be used for preventing iatrogenic injuries. In this paper the application of Navigated Control for surgery on the lateral skull base was investigated for the first time in clinical use. There was no risk structure damage with NC. Navigated Control in lateral skull base surgery seems to have a great potential for safe risk structure protection, a morbidity reduction and in a relief of strain for the surgeon. PMID- 18391281 TI - In vivo pan/tilt endoscope with integrated light source, zoom and auto-focusing. AB - We describe a surgical imaging device with pan, tilt, zoom and integrated LED light source. It can be fully inserted into the abdomen, leaving the insertion port free for tooling. Using a porcine model we have tested the device and performed surgical procedures including cholecystectomy, appendectomy, running (measuring) the bowel, suturing, and nephrectomy. The tests show that the new device is: * Easier and more intuitive to use than a standard laparoscope. * Joystick operation requires no specialized operator training. * Field of view and access to relevant regions of the body were superior to a standard laparoscope using a single port. * Time to perform procedures was better or equivalent to a standard laparoscope. We believe these insertable platforms will be an integral part of future surgical systems. PMID- 18391282 TI - On the problem of determination of spring stiffness parameters for spring-mesh models. AB - On account of having real-time behavior and being easy to implement, spring meshes have been used for modeling deformable objects. Determining spring stiffness parameters for simulation of soft objects with high accuracy still remains a challenge. Allen Van Gelder derived an approximate formula for determining spring stiffness parameters based on strain analysis. Even though the experimental result showed the effectiveness, the method has not been investigated from a quantitative point of view. In this paper we propose a quantitative method for determining spring stiffness parameters. Moreover we propose a method to improve the accuracy by way of introducing torsional spring into the conventional spring mesh model. PMID- 18391283 TI - Evaluation of the simulation robot for mandibular movements with the patient specific 3-dimensional plaster model and mandibular movement data: clinical application of the physical simulation robot. AB - We developed a multi-phase simulation system for patients with jaw deformity and dysfunction as a collaborate study between our departments. The intended application of the physical simulation robot was to evaluate its function based on well it quantitatively measured the movement of the individual patient. This physical simulation robot consists of a 6-degree-of-freedom robotic manipulator and a plaster model of patient-specific bone geometry. Each plaster model was mounted on the serial-articulated robotic manipulator. To establish the accuracy of the robot movement, the programmed movement of the robotic arm was validated using an optical tracking device. The results of the physical simulation robot corresponded with the data from the 4D analysis system. We could construct interactive relations between the 4D analysis system that was presented by virtual reality and the simulation robot which was constructed from physical simulation. PMID- 18391284 TI - MedX3D: standards enabled desktop medical 3D. AB - This paper reports on the work of the Web3D Consortium's Medical Working Group to specify and implement MedX3D -- an extension to the X3D standard that will support advanced medical visualization functionality and medical data exchange. This initiative covers volume rendering, ontology support, and data import/export, for standalone applications and web-based plug-ins. It is our hypothesis that such a 3D medical standard will provide better access to data, and enable improvements in medical care. PMID- 18391285 TI - Physics-based virtual environment for training core skills in vascular interventional radiological procedures. AB - Recent years have seen a significant increase in the use of Interventional Radiology (IR) as an alternative to open surgery. A large number of IR procedures commences with needle puncture of a vessel to insert guidewires and catheters: these clinical skills are acquired by all radiologists during training on patients, associated with some discomfort and occasionally, complications. While some visual skills can be acquired using models such as the ones used in surgery, these have limitations for IR which relies heavily on a sense of touch. Both patients and trainees would benefit from a virtual environment (VE) conveying touch sensation to realistically mimic procedures. The authors are developing a high fidelity VE providing a validated alternative to the traditional apprenticeship model used for teaching the core skills. The current version of the CRaIVE simulator combines home made software, haptic devices and commercial equipments. PMID- 18391286 TI - A cooperatively-controlled image guided robot system for skull base surgery. AB - We created an image-guided robot system to assist with skull base drilling by integrating a robot, a commercial navigation system, and an open source visualization platform. The objective of this procedure is to create a cavity in the skull base to allow access for neurosurgical interventions. The motivation for introducing an image-guided robot is to improve safety by preventing the surgeon from accidentally damaging critical structures during the drilling procedure. Our approach is to attach the cutting tool to the robot end-effector and operate the robot in a cooperative control mode, where robot motion is determined from the forces and torques applied by the surgeon. We employ "virtual fixtures" to constrain the motion of the cutting tool so that it remains in the safe zone that was defined on a preoperative CT scan. This paper presents the system design and the results of phantom and cadaveric experiments. Both experiments have demonstrated the feasibility of the system, with average overcut error at about 1 mm and maximum errors at 2.5 mm. PMID- 18391287 TI - Virtual gait training for children with cerebral palsy using the Lokomat gait orthosis. AB - The Lokomat gait orthosis was developed in the Spinal Cord Injury Center at the University Hospital Balgrist Zurich and provides automatic gait training for patients with neurological gait impairments, such as Cerebral Palsy (CP). Each patient undergoes a task-oriented Lokomat rehabilitation training program via a virtual reality setup. In four virtual scenarios, the patient is able to exercise tasks such as wading through water, playing soccer, overstepping obstacles or training in a street scenario, each task offering varying levels of difficulty. Patients provided positive feedback in reference to the utilized haptic method, specifically addressing the sufficient degree of realism. In a single case study, we verified the task difficulty. PMID- 18391288 TI - Transfer of systematic computer game training in surgical novices on performance in virtual reality image guided surgical simulators. AB - We report on a pilot study that investigates the transfer effect of systematic computer game training on performance in image guided surgery. In a group of 22 surgical novices, subjects were matched and randomized into one group training with a 3-D first person shooter (FPS) game and one group training with a 2-D non FPS game. We also included a control group. Subjects were tested pre- and post training in the MIST-VR and GI-Mentor surgical simulators. We found that subjects with past experience specific to FPS games were significantly better in performing the simulated endoscopy task, both regarding time and efficiency of screening, compared to subjects lacking FPS game experience. Furthermore subjects who underwent systematic FPS game training performed better in the MIST-VR than those training with a 2-D game. Our findings indicate a transfer effect and that experience of video games are important for training outcome in simulated surgical procedures. Video game training can become useful when designing future skills training curricula for surgeons. PMID- 18391289 TI - Initial implementation of mixed reality simulation targeting teamwork and patient safety. AB - Patient safety is one of the most pressing challenges of modern healthcare. Being a multifactorial problem, patient safety requires improvement interventions on multiple levels including individual, team and organization as a whole. Using high-fidelity human patient simulator in real clinical setting allows creating a mixed reality environment for teaching healthcare teams to improve patient safety. A multidisciplinary group of physicians at Louisiana State University Health Sciences Center in New Orleans has developed and implemented the STEPS program (System for Teamwork Effectiveness and Patient Safety) with the use of MMOR (mobile mock operating room) configuration to train general surgical teams within their own operating room environment. Each simulation session was followed by facilitated debriefing and teaching new team communication skills. Team performances were assessed by both direct observation and team's self-assessment where each team member assessed his or her own performance as well as the performance of all other team members (360 degree assessment). PMID- 18391290 TI - CT image applications for pre-surgical assessment and surgical pre-planning in Pediatric Dentistry. AB - In this study we focused on dental anomalies peculiar to children and investigated how 3D-surgical planning and simulation could be applied to each case. This study included 3 patients (2 female and 1 male, 8 to 12 years). The CT imaging was performed via a SOMATOM Plus4. From the resulting data, an image of the tooth and bone was rendered using image analysis software Amira 3.1 which was then used to reconstruct three-dimensional images. The reconstructed 3D images were imported to 3D modeling software, which provided the basis for the surgical simulations. From these results, we were able to gain important insights that helped shape the planning of the surgical operation. Furthermore we consider that these findings would be useful for the patient when taking them through the process of obtaining informed consent. PMID- 18391291 TI - Effects of expertise, practice and contextual interference on adaptations to visuo-motor misalignment. AB - The cognitive demands of endoscopic surgery are amplified when rotations of the displayed surgical field cause discrepancies between intended and actual movements. While Visuo-Motor Misalignment (VMM) often impairs surgical technical performance, motor learning literature has demonstrated adaptation acquisition with practice. It is unknown whether these findings can be generalized to complex tasks of surgery. Two experiments investigated VMM adaptation processes within a laparoscopic context. The first examined the effects of practice and angle of visual rotation while the second investigated if, and how, the Contextual Interference Effect facilitates VMM adaptations. Laparoscopic performance was positively affected with smaller angles of visual rotation (p<.001) and increasing trials (p<.001). While not significant, performance was better after blocked versus random practice. In order to facilitate VMM adaptation acquisition, it is recommended that future simulated trainers incorporate the capacity for practice under visual rotation with flexibility in practice schedule. PMID- 18391292 TI - Haptic rate for surgical manipulations with fingers and instruments. AB - In surgery, many kinds of manipulations are conducted using multiple fingers and instruments. The aim of this study is to investigate required haptic rate for multi-finger manipulation and instrumental manipulation. This paper investigated required haptic rate for multi-finger haptic interaction with haptic device and soft tissue deformation. Results of experiment clarified the fact that there is difference of threshold of haptic rate between the number of manipulating fingers. PMID- 18391293 TI - Robotic telepresence for collaborative clinical outreach. AB - The increasing complexity of healthcare and shortage of clinical specialists needs to be addressed through communication, collaboration and coordination of resources to ensure timely delivery of clinical expertise. Remote Presence is a next generation telemedicine technology platform which combines the power of robotics, wireless, and the internet to enable hospitals and physicians to bring the right care to the right patient at the right time. For example, Remote Presence has been successfully implemented in a hub and spoke model allowing stroke neurologists at a stroke center of excellence to provide the spoke hospital staff with patient consultation and training services. The results are improved geographical reach of stroke specialist care throughout the region with significant impact on patient outcomes as well as improved alignment with established care standards and best practices. PMID- 18391294 TI - In vivo robotics for natural orifice transgastric peritoneoscopy. AB - Natural Orifice Translumenal Endoscopic Surgery (NOTES) is potentially the next paradigm shift in minimally invasive surgery. Currently, NOTES procedures are performed using modified endoscopic tools with significant constraints. New tools are necessary that allow the surgeon to better visualize and dexterously manipulate within the surgical environment. In this study, a two-armed dexterous miniature in vivo robot with stereoscopic vision capabilities has been developed that addresses many of these constraints. The design and kinematic configuration of the robot allows for its complete insertion into the peritoneal cavity, and provides intuitive visualization and sufficient force application for tissue manipulation within the dexterous workspace. The NOTES robot successfully demonstrated various capabilities in a non-survival natural orifice surgical procedure in a porcine model suggesting the feasibility of using miniature in vivo robots for performing natural orifice procedures within the peritoneal cavity. PMID- 18391295 TI - VR robotic surgery: randomized blinded study of the dV-Trainer robotic simulator. AB - This research represents a randomized blinded pilot study to evaluate the acceptability and validity of a da Vinci robotic virtual reality simulator platform tested during a pediatric robotic surgery post-graduate course during the annual American Urological Association meeting in June 2007. Course enrollees performed robotic skills tasks on the da Vinci robot and on an offline dV-Trainer and course participant demographic and performance data were analyzed. The majority of learners believed that VR simulation is useful for teaching robotic skills, they believed that the offline trainer can teach robotic skills comparable to a dry lab robotics skills station, and the offline trainer was able to discriminate between experts and novices of robotic surgery, thereby meeting criteria for face, content, and construct validities. This is the first reported acceptability study of a VR robotic surgery simulator as compared to the da Vinci robot system. PMID- 18391296 TI - The biomechanics of percutaneous needle insertion. AB - Emphasis has been placed on improving patient outcomes in healthcare management. Significant patient morbidity and mortality exists from inappropriate procedural technique and percutaneous catheter needle insertion procedures have been linked to medical complications. Healthcare trainees learn these procedures through trial and error and most existing simulators are synthetic tissue based and lack in-vivo force feedback. We seek to utilize the Blue DRAGON instrument positioning system coupled with a force sensor to determine true forces experienced by a needle as it is passed through animal and human tissues in an effort to design a percutaneous needle insertion simulator that affords the learner with the experience of the true force feedback. Acquiring force displacement measurements of needle insertion is the first step towards development of a computational model of the phenomena. The computational model may be further incorporated into a medical haptic simulator that provides physically based force feedback to the user. PMID- 18391297 TI - MR fluid haptic system for regional anesthesia training simulation system. AB - Magneto-Rheological (MR) fluid is a class of controllable smart material whose rheological properties may be rapidly varied by the application of a magnetic field. In this paper we focus on our MR fluid based haptic system providing accurate haptic feedback for needle insertion and syringe injection. We use the essential characteristic of MR fluid, a controllable yielding strength in milliseconds under magnetic field, in our innovative MR fluids-based haptic feedback device that imposes simulated force for the regional anesthesia simulator. PMID- 18391298 TI - Soft-tissue characterization during monopolar electrocautery procedures. AB - Advances in electrosurgical technology have promoted the use of electrocautery in many surgical procedures. Precise modeling of soft tissue deformation during electrocautery with electrosurgical generators can be a valuable tool in training simulators for surgical procedures. Coupling the visualization of electrocautery with the force feedback during an electrocautery process (to maintain optimal current) without causing necrosis is an important learning tool. Realistic simulation will provide surgeon trainees a method to practice electrocautery techniques prior to experimenting on live tissue as well as allow surgeons to gain a feel for electrocautery procedures. PMID- 18391299 TI - Representing fluid with smoothed particle hydrodynamics in a cranial base simulator. AB - We describe the implementation of irrigation and blood simulation using Smoothed Particle Hydrodynamics (SPH) in a cranial base surgical simulator. Graphical accuracy of virtual surgery is a significant goal for improving the realism and immersive experience of computerized training environments. For temporal bone micro-surgery fluids contribute not only to the visual integrity of the surgical field but provide relevant anatomic cues as well. The skill of 3-D sensory and navigation has become increasingly viable in surgery with the rising popularity of laparoscopic, catheter angiography and other minimally invasive approaches. The introduction of realistic simulated blood and irrigation enables the practice and coordination of two-handed microdissection techniques and the timing needed for safe bone removal and cautery. PMID- 18391300 TI - CUDA-based real time surgery simulation. AB - In this paper we present a general software platform that enables real time surgery simulation on the newly available compute unified device architecture (CUDA)from NVIDIA. CUDA-enabled GPUs harness the power of 128 processors which allow data parallel computations. Compared to the previous GPGPU, it is significantly more flexible with a C language interface. We report implementation of both collision detection and consequent deformation computation algorithms. Our test results indicate that the CUDA enables a twenty times speedup for collision detection and about fifteen times speedup for deformation computation on an Intel Core 2 Quad 2.66 GHz machine with GeForce 8800 GTX. PMID- 18391301 TI - Objective assessment of telesurgical robot systems: Telerobotic FLS. AB - The Society of American Gastrointestinal Endoscopic Surgeons (SAGES) Fundamentals of Laparoscopic Surgery (FLS) program contains curriculum that includes both a cognitive and psychomotor skills. In this research the use of FLS Block Transfer task is used to evaluate the performance of surgeons' teleoperating the University of Washington Surgical robot. The use of the FLS Trainer Box and accessories kit provides a well-defined series of tasks that can be repeated by any researchers working in the field of surgical robotics so that systems can be evaluated using a common method. PMID- 18391302 TI - An efficient dynamic point algorithm for line-based collision detection in real time surgery simulation involving haptics. AB - In this paper, we introduce a novel "dynamic point" algorithm for computing the interaction of a line-shaped haptic cursor and polygonal surface models which has a near constant complexity. The algorithm is applied in laparoscopic surgery simulation for interaction of surgical instruments with physics-based deformable organ models. PMID- 18391303 TI - Physics-based real time laparoscopic electrosurgery simulation. AB - While physics-based modeling of electrosurgical procedures is essential for most laparoscopic simulation systems, we present such a system for the first time in this paper. We have implemented a physics-based model of electrosurgery to control the temperature distribution on the tissue as a function of time. Then, we evaluate the algorithm within a complete graphics-haptics-physics-based system. PMID- 18391304 TI - Towards a virtual basic laparoscopic skill trainer (VBLaST). AB - Surgical skill training is a long and tedious process of acquiring fine motor skills. To overcome the drawbacks of the existing toolbox trainer systems, we develop, for the first time, a virtual basic laparoscopic skill trainer (VBLaST) whereby tasks, such as the ones available in the FLS toolbox system, may be performed on the computer. PMID- 18391305 TI - Revealing the conceptual substrate of biomedical cognitive models to the wider community. AB - The patient authoring interface for each disease in the Maryland Virtual Patient simulation environment reveals the conceptual substrate of the disease model. Revealing the disease model to the community both explains how the interactive simulations work and invites collaboration from the wider community. PMID- 18391306 TI - The emergence of multi-modal distraction as a paediatric pain management tool. AB - The emergence of Multi Modal Distraction (MMD) has occurred following appraisal of the developmental and clinical concerns surrounding Virtual Reality's (VR) applicability to pediatric pain management. MMD was developed to expand current distraction tools and technology available for pain management in the pediatric population. This paper will examine how the challenges faced by VR have motivated the collaborative design and development process of MMD. It will then outline the continuous clinical trialling being undertaken to ensure its efficacy in managing childhood procedural anxiety and pain. PMID- 18391307 TI - Physically valid surgical simulators: linear versus nonlinear tissue models. AB - Realistic modeling of the interaction between surgical instruments and organs has been recognized as a key requirement in the development of high-fidelity surgical simulators. For a nonlinear model, the well-known Poynting effect developed during shearing of the tissue results in normal forces not seen in a linear elastic model. It is demonstrated that the difference in force magnitude for myocardial tissue is larger than the just noticeable difference for contact force discrimination thresholds published in the psychophysics literature. This work also proposes the validation of simulators by careful examination of relevant simulator design parameters that relate to final simulator behaviors affecting clinical outcomes. PMID- 18391308 TI - Computer integrated system for minimally invasive lung brachytherapy. AB - This paper describes a novel approach for lung cancer treatment. It accesses the lung through small incisions in a minimally invasive manner to insert radioactive seeds into the tumour using a computer integrated system. A test-bed incorporating robotic assistance, electromagnetic tracking and ultrasound imaging has been developed. Upgrades to the navigational software, show a considerable improvement in performance when compared to an earlier system. PMID- 18391309 TI - Verifying the effectiveness of a computer-aided navigation system for arthroscopic hip surgery. AB - Computer-aided technology can decrease the difficulty associated with arthroscopic surgery. Unlike a larger incision that exposes the whole joint, a small arthroscopic incision limits the surgeon to view the joint only from the arthroscope. Our computer-aided system for hip arthroscopy addresses this loss of joint visibility by (1) tracking tool position with a linkage of encoders, and (2) indicating the tool position relative to the patient anatomy. This paper presents a study of user performance to verify the effectiveness of this computer aided system for hip arthroscopy. A user study was completed to determine if the computer-aided system could help reduce task completion time and tool path length. Ten participants completed a simple navigation task with and without the assistance of the computer-aided system. A time reduction of 38% and a 72% tool path-length decrease was achieved with the computer-aided system, confirming its effectiveness. PMID- 18391310 TI - Barriers to wider adoption of mobile telerobotic surgery: engineering, clinical and business challenges. AB - A portable robotic telesurgery network could remove the geographic disparity of surgical care and provide expert surgical support for first responders to traumatic injury. This is particularly relevant to battlefield medicine where surgical intervention is currently not available to the most perilous fighting circumstances. Similar utility applies to the peacetime healthcare mission. The authors identify the potential advantage to healthcare from a mobile robotic telesurgery system and specify barriers to the employability and acceptance of such a system. The proposed research roadmap will describe a portable telesurgery system that represe government/industrial recognition and reward for excellent care provided by quality surgeons. The provision of expert surgical care improves the outcomes of surgical intervention by reducing errors. For example, during the common procedure of laparoscopic cholecystectomy, distributed telesurgical care could normalize surgical performance and limit major variance of surgeon outliers; such as reducing common bile duct injury to the very low rate seen with operation by proficient surgeons. PMID- 18391311 TI - Real-time blood vessel deformation with bleeding based on particle method. AB - Many surgical simulators have been developed in various fields; however, there is little consideration about the simulation of blood vessel deformation and bleeding, which is very important for almost all surgical simulators. This paper describes the method how to deform a blood vessel very fast and stably, and to simulate bleeding at the same time. As a result, we could perform the concurrent representation of blood vessel deformation and bleeding in real time with a normal PC. PMID- 18391312 TI - Interactive volume manipulation for supporting preoperative planning. AB - This paper presents a volume manipulation framework by which surgeons can interactively manipulate soft models like through surgical tools. The framework robustly simulates common surgical manipulations such as grasping, holding, cutting and retraction. We simulate cutting based on FEM formulation by replacing vertices and eliminating elements, without subdividing elements or adding new vertices. The size of stiffness matrix is constant. We also present real-time volume shading methods for deformable modeling. Our algorithms achieved interactive response in volume manipulation. Several surgical approaches and procedures were rehearsed and used for preoperative discussion. PMID- 18391313 TI - Modeling surgical tool selection patterns as a "traveling salesman problem" for optimizing a modular surgical tool system. AB - As modular systems come into the forefront of robotic telesurgery, streamlining the process of selecting surgical tools becomes an important consideration. This paper presents a method for optimal queuing of tools in modular surgical tool systems, based on patterns in tool-use sequences, in order to minimize time spent changing tools. The solution approach is to model the set of tools as a graph, with tool-change frequency expressed as edge weights in the graph, and to solve the Traveling Salesman Problem for the graph. In a set of simulations, this method has shown superior performance at optimizing tool arrangements for streamlining surgical procedures. PMID- 18391314 TI - GPU accelerated viscous-fluid deformable registration for radiotherapy. AB - Repeated acquisitions of computed tomography and magnetic resonance imaging are increasingly used during radiotherapy treatment to accurately deliver radiation while limiting side effects. This is only feasible however after all acquisitions have been correlated to a single reference scan using a deformable registration method. This paper presents a parallel implementation of one such method, the viscous-fluid registration method, on modern graphics hardware. A significant speedup close to two orders of magnitudes was observed when comparing to a CPU based implementation. As a consequence of the reduced registration time it is now feasible to perform larger scale clinical evaluation of the method. An example of registration results obtained during a treatment course is included in the paper. PMID- 18391315 TI - A framework for shape matching in deformable image registration. AB - Many existing image registration methods have difficulties in accurately describing significant rotation and bending of entities (e.g. organs) between two datasets. A common problem in this case is to ensure that the resulting registration is physically plausible, i.e. that the registration describes the actual bending/rotation occurring rather than just introducing expansion in some areas and shrinkage in others. In this work we developed a general framework for deformable image registration of two 3D datasets that alleviates this problem. To ensure that only physically feasible and plausible solutions to the registration problem are found, a soft tissue deformable model is used to constrain the search space for the desired correspondence map while minimizing a similarity metric between the source and reference datasets. Results from a deformable phantom experiment were used to verify and evaluate the framework. PMID- 18391316 TI - Optimizing joint placement and motion schedule for 2-DOF computer-assisted distraction osteogenesis. AB - Distraction osteogenesis is a surgical procedure used to correct severe bone deformities. Current practice involves planning the deformity correction using parameters measured from radiographs and the installation of complex fixators such as the Taylor spatial frame. A computer-integrated surgery system has been proposed that uses a novel two-degree-of-freedom fixator to correct complex bone deformities. The proposed fixator is designed such that the two joints can be positioned and oriented arbitrarily to achieve proper alignment of the bone fragments while following a desired trajectory. This paper describes the fixator configuration methods and trajectory planning procedures used to prove the feasibility of such a device. It is shown that this device is applicable for a variety of deformity types and realignment trajectories. PMID- 18391317 TI - Hip motion analysis using multi phase (virtual and physical) simulation of the patient-specific hip joint dynamics. AB - In total hip arthroplasty (THA), the patient-specific bone geometry or the characteristics of the skeletal movement should be considered during treatment in order to prevent complications. In this paper, we propose a novel approach for the analysis of joints which combines the patient-specific virtual and physical simulation. The patient-specific anatomical structure and hip motion was obtained from CT and optical motion capture. The virtual simulation was conducted by integrating these data using virtual reality technique. The physical simulation was achieved by using plaster models of the patient's pelvis and femur and robotic manipulator. The plaster models were driven by two robotic manipulators to reproduce the hip motion. The accuracy of the robot movement was 0.245 mm over the working area according to the validation by an optical tracking system. By combining this system with linear actuators that reproduce the muscle functions, patient-specific muscle function can be simulated, thereby helping clinicians to diagnose and make a treatment plan. PMID- 18391318 TI - Simulation debriefing and quantitative analysis using video analysis software. AB - Coding of simulation video provides real benefits during debriefing. We demonstrate examples of the use of software to quantify events in video. PMID- 18391319 TI - Transforming an educational virtual reality simulation into a work of fine art. AB - This paper outlines user interface and interaction issues, technical considerations, and problems encountered in transforming an educational VR simulation of a reified kidney nephron into an interactive artwork appropriate for a fine arts museum. PMID- 18391320 TI - Virtual environment for assessment of neurocognitive functioning: virtual reality cognitive performance assessment test. AB - While standard neuropsychological measures have been found to have adequate predictive value, their ecological validity may diminish predictions about real world functioning. Virtual environments (VEs) are increasingly recognized as ecologically valid tools for neuropsychological assessment. We aim to develop a VE-based neuropsychological battery delivered within the context of a virtual city: Virtual Reality Cognitive Performance Assessment Test (VRCPAT). The 15 minute VRCPAT battery and 1.5 hour in-person neuropsychological assessment were conducted with a sample of 40 healthy adults, between the ages of 21 and 36, that included equivalent distributions of men and women from ethnically diverse populations. No subjects had history of psychiatric or neurologic conditions. Results supported both convergent and discriminant validity. The VRCPAT was found to correlate significantly with traditional neuropsychological tests assessing similar target constructs. No significant correlations existed between VRCPAT measures and non-target constructs. Findings suggest that the VRCPAT measures a capacity consistent with that of traditional neurocognitive measures; and is inconsistent with potential confounds. We conclude that the VRCPAT provides a unique opportunity to reliably and efficiently study neurocognitive function within an ecologically valid environment. PMID- 18391321 TI - Objective structured clinical interview training using a virtual human patient. AB - Effective interview skills are a core competency for psychiatry residents and developing psychotherapists. Although schools commonly make use of standardized patients to teach interview skills, the diversity of the scenarios standardized patients can characterize is limited by availability of human actors. Further, there is the economic concern related to the time and money needed to train standardized patients. Perhaps most damaging is the "standardization" of standardized patients -- will they in fact consistently proffer psychometrically reliable and valid interactions with the training clinicians. Virtual Human Agent (VHA) technology has evolved to a point where researchers may begin developing mental health applications that make use of virtual reality patients. The work presented here is a preliminary attempt at what we believe to be a large application area. Herein we describe an ongoing study of our virtual patients (VP). We present an approach that allows novice mental health clinicians to conduct an interview with a virtual character that emulates an adolescent male with conduct disorder. This study illustrates the ways in which a variety of core research components developed at the University of Southern California facilitates the rapid development of mental health applications. PMID- 18391322 TI - Our experience with virtual craniomaxillofacial surgery: planning, transference and validation. AB - Clinical application of virtual craniomaxillofacial surgery (VCMS) planning is demonstrated with four typical cases. An integral component to success is transferring the surgical plan to the operating environment within a reasonable time frame through the design and fabrication of surgical guides and implants. PMID- 18391323 TI - Virtual reality training for radiotherapy becomes a reality. AB - A report in 2007 to the UK Government identified a crisis in England for training staff and students for the radiotherapy treatment of cancer. The Hull authors have developed an immersive life size virtual environment of a radiotherapy treatment room, known as VERT, to address this problem. VERT provides the trainee with models, simulation, enhanced visualization and training aids for treatment of virtual patients in a virtual treatment room. In 2007 immersive VERT systems for radiotherapy training were established for training purposes at the University Aarhus Hospital (Denmark) and the Birmingham City University (UK). This paper reports on early evaluations of VERT by these two institutions. PMID- 18391324 TI - Comparative usability studies of full vs. partial immersive virtual reality simulation for medical education and training. AB - Virtual reality (VR) simulation provides a means of making experiential learning reproducible and reusable. This study was designed to determine the efficiency and satisfaction components of usability. Previously, it was found that first year medical students using a VR simulation for medical education demonstrated effectiveness in learning as measured by knowledge structure improvements both with and without a head mounted display (HMD) but students using a HMD showed statistically greater improvement in knowledge structures compared to those not using a HMD. However, in this current analysis of other components of usability, there were no overall significance differences in efficiency (ease of use), nor in satisfaction, within this same group of randomized subjects comparing students using a HMD to those not using a HMD. These types of studies may be important in determining the most appropriate, cost effective VR simulation technology needed to achieve specific learning goals and objectives. PMID- 18391325 TI - Interactive finite element simulation of the beating heart for image-guided robotic cardiac surgery. AB - An interactive finite element simulation of the beating heart is described in which the intrinsic motion is implied from preoperative 4D tomographic scan data. The equations of motion are reversed such that, given changes in node displacements over time, the node forces that produce those changes are recovered. Subsequently, these forces are resolved from the global coordinate system into systems local to each mesh element such that, at each simulation time step, the collection of node forces can be expressed as simple weighted sums of current node positions. This facilitates the combination of extrinsic forces like those due to tool-tissue interactions, gravity, insufflation of the thoracic cavity and left lung deflation. The method has been applied initially to volumetric images of a pneumatically-operated beating heart phantom. PMID- 18391326 TI - Fatty tissue in a haptic illustration environment. AB - Modeling soft tissue for surgery simulation is a challenging task due to the complex way that the tissue can deform and interact with virtual surgical tools manipulated by user. One soft tissue that is ubiquitous but often not modeled, is fatty tissue. Here we present a novel fatty tissue model based on the mass-spring system on the Graphics Processing Unit (GPU) as part of our Toolkit for Illustration of Procedures in Surgery (TIPS). The user can interact with the fatty tissue in real time via a handheld haptic stylus that represents a virtual surgical tool in TIPS environment. The currently available interactions are palpation, grasp, and cut. PMID- 18391327 TI - An adaptive transmission protocol for managing dynamic shared states in collaborative surgical simulation. AB - A force prediction algorithm is proposed to facilitate virtual-reality (VR) based collaborative surgical simulation by reducing the effect of network latencies. State regeneration is used to correct the estimated prediction. This algorithm is incorporated into an adaptive transmission protocol in which auxiliary features such as view synchronization and coupling control are equipped to ensure the system consistency. We implemented this protocol using multi-threaded technique on a cluster-based network architecture. PMID- 18391328 TI - Visualization of biomechanical properties of female pelvic floor function using video motion tracking of ultrasound imaging. AB - Real time ultrasound imaging is one of the many ways to clinically evaluate the anatomical and functional condition of female pelvic floor in patients with urinary incontinence. Reflex arc testing of the displacement of uro-gynecological structures during imaging provides a non-invasive way of visualizing their motility. The response from such tests invariably contains a very large amount of visual information, which is not readily captured and assimilated by the observer because it occurs so fast. For this reason only a portion of available information is retained, typically the beginning end ending frame of the image is preserved. Using video motion tracking, parameters were identified to define important biomechanical and temporal relationships. We conclude that new and clinically significant amount of original information about the female pelvic floor can be obtained through the proposed analysis of visualizations. PMID- 18391329 TI - Automatic recognition of surgical motions using statistical modeling for capturing variability. AB - The ability to accurately recognize elementary surgical gestures is a stepping stone to automated surgical assessment and surgical training. However, as the pool of subjects increases, variation in surgical techniques and unanticipated motion increases the challenge of creating robust statistical models of gestures. This paper examines the applicability of advanced modeling techniques from automated speech recognition to the problem of increasing variability in surgical motions. In particular, we demonstrate the effectiveness of automatically bootstrapped user-adaptive models on diverse data acquired from the da Vinci surgical robot. PMID- 18391330 TI - Creating an animatable 3D volume hand model from in vivo MRI. AB - Volume graphics has obvious benefits to medical visualization, since it represents the complete 3D information of both surface appearance and the underlying anatomical structures. This study presents an approach to rapidly creating an animatable 3D volume from in vivo human hand MRI scans. The result is a fully articulated hand volume driven by intuitive joint control that respects rigid deformation of the bone structures and produces smooth deformations of both the skin surface and the interior soft tissue regions. While the method can potentially be applied to any articulated body region, the human hand is chosen to illustrate the process, both due to its intrinsic interest in medical applications and because of the large number of degrees of freedom and challenging anatomy of the hand. PMID- 18391331 TI - Haptic feedback for multilayer cutting. AB - An approach in effectively estimating the force feedback for a tactile haptic based on multi-proxy rendering for 3D surface cuttings for a virtual surgery simulation is described in this paper. The force-models representing haptic force feedback are approximated using D'Alembert's principle in the mechanic case of spring-damper-stiffness interaction of the surfaces. We also propose a combination between mesh refinement and adaptive re-meshing to create a progressive cutting over the layering surfaces. Experimental results prove that the physical interaction to create cutting paths over the multilayer surfaces can be deliver smoothly with haptic in real time with 3D visual stereo on a PC. PMID- 18391332 TI - Asynchronous teaching of psychomotor skills through VR annotations: evaluation in digital rectal examination. AB - Many VR technology based training systems use expert's motion data as the training aid, but would not provide any short-cut to teaching medical skills that do not depend on exact motions. Earlier we presented Annotated Simulation Records (ASRs), which can be used to encapsulate experts' insight on psychomotor skills. Annotations made to behavioural parameters in training simulators enable asynchronous teaching instead of just motion training in a proactive way to the learner. We evaluated ASRs for asynchronous teaching of Digital Rectal Examination (DRE) with 3 urologists and 8 medical students. The ASRs were found more effective than motion-based training with verbal feedback. PMID- 18391333 TI - Why you really eat? Virtual reality in the treatment of obese emotional eaters. AB - The aim of this study is to assess the efficacy of a specific stress management protocol, based on immersive Virtual Reality (VR), to be used as part of a multidisciplinary inpatient program for the treatment of obesity. The stress management protocol included imagery, relaxation and different cognitive behavioral approaches -- emotion focused coping, self-monitoring and record keeping. Forty (40) participants, all female, were recruited for the study at the San Giuseppe Hospital, Istituto Auxologico Italiano, Piancavallo (VB), Italy. Participants were chosen among the obese patients without a specific Binge Eating diagnosis but with high level of anxiety and a history of emotional eating. The sample was randomly distributed in three conditions as follows: (a) VR stress Management Protocol; (b) DVD based stress management protocol; (c) no treatment (control condition). Data show that the VR condition produced a significantly higher reduction in anxiety, as compared both to DVD and control groups. More, we found a significant correlation between changes in the emotional state and the level of presence: the more present the users felt, the higher the reduction in anxiety. This datum suggests the possible role of presence in mediating the effects of a stress management protocol. PMID- 18391334 TI - Virtual Iraq: initial results from a VR exposure therapy application for combat related PTSD. AB - Post Traumatic Stress Disorder (PTSD) is reported to be caused by traumatic events that are outside the range of usual human experience including (but not limited to) military combat, violent personal assault, being kidnapped or taken hostage and terrorist attacks. Initial data suggests that at least 1 out of 6 Iraq War veterans are exhibiting symptoms of depression, anxiety and PTSD. Virtual Reality (VR) delivered exposure therapy for PTSD has been used with reports of positive outcomes. The aim of the current paper is to present the rationale and brief description of a Virtual Iraq PTSD VR therapy application and present initial findings from its use with PTSD patients. Thus far, Virtual Iraq consists of a series of customizable virtual scenarios designed to represent relevant Middle Eastern VR contexts for exposure therapy, including a city and desert road convoy environment. User-centered design feedback needed to iteratively evolve the system was gathered from returning Iraq War veterans in the USA and from a system deployed in Iraq and tested by an Army Combat Stress Control Team. Clinical trials are currently underway at Ft. Lewis, Camp Pendleton, Emory University, Weill Cornell Medical College, Walter Reed Army Medical Center, San Diego Naval Medical Center and 12 other sites. PMID- 18391335 TI - Collaborative engineering: 3-D optical imaging and gas exchange simulation of in vitro alveolar constructs. AB - This paper reports on the computational simulation and modeling of an in vitro alveolar construct system along the optical coherence microscopy (OCM) methods for visualizing engineered tissue. The optical imaging methods will be compared to immunohistochemical light microscopy samples of engineered alveolar constructs. Results show depth images of the alveolar tissue construct for a bilayer construct, as well as predictions of the gas exchange process in a simple model of a bio-reactor hosting the construct. PMID- 18391336 TI - A virtual reality throat examination simulation. AB - Computer based patient simulators can provide an alternative to inanimate clinical skills training. Here we present a virtual reality throat examination simulation capable of simulating a range of throat pathologies. The computer interface employs haptic feedback to allow students to interact naturally with the software. PMID- 18391337 TI - A virtual reality 3D jigsaw for teaching anatomy. AB - Virtual Reality has some advantages over traditional teaching and learning media. Here we describe a VR Jigsaw which uses a novel interface to facilitate learning the anatomy of the skull. A small trial was performed which indicates that the software succeeds at engaging students and suggests that their comprehension of complex 3D structures was improved. PMID- 18391338 TI - Monitoring simulated maternal-fetal heart rate signals. AB - The present study represents the first in a series aimed at examining how individuals monitor maternal-fetal heart rate signals over an extended period of time. Seventy-two undergraduate students monitored simulated maternal-fetal heart rate signals for late decelerations of either high or low amplitude for 48 minutes. The results showed that the ability to detect late decelerations was quite good and did not decline over time. Although the findings regarding detections are encouraging, the participants also made a lot of false alarms. Collectively, these results suggest that the observers still struggled to distinguish critical patterns from the background fetal heart activity. PMID- 18391339 TI - Open-source shared case library. AB - Sharing simulator case scenarios among educators will greatly reduce the cost and effort involved in bringing simulation to our students. A library of 72 cases was posted to the internet to freely share. The cases in the library consist of text files utilizing a simple open-source format. In the first four months after posting, cases were downloaded over 18,000 times. In addition, the library encouraged the development of 23 new cases by 16 new contributors. This experience indicates a strong interest in obtaining free cases, but only modest willingness to share new scenarios among authors and institutions. PMID- 18391340 TI - Integration of virtual reality and conventional skills trainers: a mixed resource model. AB - Training surgical skills in virtual reality (VR) trainers holds promise for successful skill acquisition. VR can simulate complex procedures, present varied physiology, and provide detailed performance feedback. However, VR trainers have not gained wide acceptance as a dominant training modality. Simple mechanical "box trainers" are still used more widely for training basic skills, with the possibility of training more complex skills using more sophisticated physical models of physiology. Combining both VR and mechanical box trainers in a unified curriculum can capitalize on the strengths of VR and the strengths of simple mechanical trainers in an integrated program of practice and feedback. Diagnostics from performance on VR trainers can be used to develop a personalized curriculum for practice on box trainers, with detailed feedback provided through intermittent VR sessions. This integrated approach can increase resource utilization, improve training efficiency, and may lead to better transfer of training to the patient-care environment. PMID- 18391341 TI - Simulation of punch biopsies: a case study. AB - As the incidence of skin cancer continues to rise, there is an increasing need for skilled practitioners that are proficient in identifying suspicious lesions and competent in acquiring biopsies that provide for optimal determination of malignancy and staging. We report on the development of a prototype simulation that emulates the basic procedures necessary to acquire a punch biopsy. The objective of this effort is to produce a low-cost, effective method to teach non specialists, i.e., nurse practitioners, internists, etc., the optimal placement of the punch to obtain a biopsy for pathological analysis. The simulation can be utilized for synchronous sessions with remote experts, as well as asynchronous sessions for deliberate practice. The simulation is designed to allow for easy import of digital images of various lesions to promote use and present the wide pathological variance experienced in the clinic. PMID- 18391342 TI - Haptic guided laparoscopy simulation improves learning curve. AB - Haptic fidelity has been increasingly emphasized in MIS simulators however the real benefits of the simulated environments are mostly under-exploited for MIS Simulation and Training. In this paper we discuss the use of augmented forces such as haptic guidance in our laparoscopic simulator for improving learning curve. A simple virtual environment has been designed to simulate advanced MIS tasks such as cutting. A simple haptic guidance system has been implemented to provide a guiding force pulling a novice trainee along the ideal motion path for manipulating the laparoscopic tools. An experiment has been designed to compare the time taken by two groups of participants with and without guidance for the cutting procedure. The results indicate that the haptic guidance has assisted participants in learning of motor skills in its cognitive and associative stage of learning. PMID- 18391343 TI - Realistic soft tissue deformation strategies for real time surgery simulation. AB - A volume-preserving deformation method (VPDM) is developed in complement with the mass-spring method (MSM) to improve the deformation quality of the MSM to model soft tissue in surgical simulation. This method can also be implemented as a stand-alone model. The proposed VPDM satisfies the Newton's laws of motion by obtaining the resultant vectors form an equilibrium condition. The proposed method has been tested in virtual surgery systems with haptic rendering demands. PMID- 18391344 TI - Changes in three dimensional simulation models of the airway which are due to increases in age or body mass index. AB - The aim of this study was to model the changes in the soft and hard tissues that occur around the upper airway with increasing age and weight, and to visually gauge them through the use of 3D simulation models. We created two standard 3 dimensional models, with one set to represent a healthy subject model and the other serving as an OSA model. The results of the regression models in our anatomical variables were attached to each 3D model. We compared our data with the data of previous studies to recognize the validity of our regression model. In both models the pharyngeal length increased as age increased. We observed an increase in the fat tissue, soft palate length, and a thickening of the soft palate as BMI increased. As age and BMI increased, the narrowing of the airway appeared more severe in the OSA model. PMID- 18391346 TI - Comparative study of haptic training versus visual training for kinesthetic navigation tasks. AB - Kinesthetic motion appears in tasks ranging from minimally invasive surgical procedures to patient rehabilitation. In this work, a comparative study is performed using two training paradigms for kinesthetic tasks. Subjects are trained to learn a complex 3D path through either the haptic method or the visual method. After the training period, subjects trace the learned 3D path without any feedback. Performance is evaluated primarily based on path deviation and time. Results indicate that haptically trained users have significantly higher performance than visually trained users. Other relevant results are also presented that can have a significant effect in the design of haptics-based interaction systems. PMID- 18391345 TI - Multi-parallel open technology to enable collaborative volume visualization: how to create global immersive virtual anatomy classrooms. AB - Many prototype projects aspire to develop a sustainable model of immersive radiological volume visualization for virtual anatomic education. Some have focused on distributed or parallel architectures. However, very few, if any others, have combined multi-location, multi-directional, multi-stream sharing of video, audio, desktop applications, and parallel stereo volume rendering, to converge on an open, globally scalable, and inexpensive collaborative architecture and implementation method for anatomic teaching using radiological volumes. We have focused our efforts on bringing this all together for several years. We outline here the technology we're making available to the open source community and a system implementation suggestion for how to create global immersive virtual anatomy classrooms. With the releases of Access Grid 3.1 and our parallel stereo volume rendering code, inexpensive globally scalable technology is available to enable collaborative volume visualization upon an award-winning framework. Based upon these technologies, immersive virtual anatomy classrooms that share educational or clinical principles can be constructed with the setup described with moderate technological expertise and global scalability. PMID- 18391347 TI - Towards THz medical imaging; reflective imaging of animal tissues. AB - A reflective THz imaging system has been developed, and features a photoconductive switch and zero-bias Schottky diode detector. The system was used to image deli meats and can distinguish between muscle and adipose tissue based on water content. This capability is a step towards the development of THz medical imaging systems. PMID- 18391348 TI - The distortion of reality perception in schizophrenia patients, as measured in Virtual Reality. AB - BACKGROUND: Virtual Reality is an interactive three-dimensional computer generated environment. Providing a complex and multi-modal environment, VR can be particularly useful for the study of complex cognitive functions and brain disorders. Here we used a VR world to measure the distortion in reality perception in schizophrenia patients. METHODS: 43 schizophrenia patients and 29 healthy controls navigated in a VR environment and were asked to detect incoherencies, such as a cat barking or a tree with red leaves. RESULTS: Whereas the healthy participants reliably detected incoherencies in the virtual experience, 88% of the patients failed in this task. The patients group had specific difficulty in the detection of audio-visual incoherencies; this was significantly correlated with the hallucinations score of the PANSS. CONCLUSIONS: By measuring the distortion in reality perception in schizophrenia patients, we demonstrated that Virtual Reality can serve as a powerful experimental tool to study complex cognitive processes. PMID- 18391349 TI - SimTools: a new paradigm in high fidelity simulation. AB - We demonstrate a prototype set of clinical devices that deliver simulated information to the clinician when used with standardized patients or simple manikins. PMID- 18391350 TI - Virtual patient monitors for new user familiarization. AB - We demonstrate web-based ECG monitor simulators that enable students to gain and demonstrate competence with equipment specific to a particular workplace. PMID- 18391351 TI - Virtual standardized patients for training health professionals on alcohol screening and brief intervention. AB - Alcohol use disorders are common, but are often not discussed with patients. Through NIAAA funding, SIMmersion LLC and the University of Wisconsin are producing an interactive, immersive computer-based simulation that will allow learners to conduct an alcohol screening and brief intervention with a virtual standardized patient. The validity of this simulation will be evaluated through a study involving 100 clinicians. PMID- 18391352 TI - Virtual endoscopy on a portable navigation system for ENT surgery. AB - Virtual endoscopy is considered as an ideal aid assessing the complex anatomy of patients and has already been evaluated in several clinical studies. However, due to the increasing quality of modern CT- and MRT-images, present virtual-endoscopy software relies on powerful hardware. In this contribution virtual endoscopy on a portable navigation system for ENT surgery is proposed. The portable navigation system features a tablet pc to ensure a device that does not need much space in the operating room. PMID- 18391353 TI - Model for nerve visualization in preoperative image data based on intraoperatively gained EMG signals. AB - While removing bone tissue of the mastoid, the facial nerve is at risk of being injured. In this contribution a model for nerve visualization in preoperative image data based on intraoperatively gained EMG signals is proposed. A neuro monitor can assist the surgeon locating and preserving the nerve. With the proposed model gained EMG signals can be spatially related to the patient resp. the image data. During navigation the detected nerve course will be visualized and hence permanently available for assessing the situs. PMID- 18391354 TI - Evaluating elicited anxiety in a simulated environment. AB - Simulated environments are increasingly being used to evaluate human behavior in a wide variety of situations. These virtual environments are particularly useful in the study of behavior involving dangerous conditions. Quantifying the impact of these environments on the subject is essential to validate the efficacy of the simulation as a virtualization of reality. We have developed a prototypical environment that simulates a common agricultural setting that provides the potential for acute injuries. We report on an exploratory study to evaluate the efficacy of our design to illicit a sense of realism and subsequently evoke a physiological response of anxiety in the subject. We present on the components that comprise the environment, our study design, and preliminary results. PMID- 18391355 TI - Translating human simulation technologies to veterinary surgical training: accelerating adoption. AB - Through the reduction of live animal use in teaching surgical technique, the opportunities to deliberately study complex regional anatomy and practice surgical technique have decreased. With reduced exposure, there is concern some individuals are graduating without the requisite knowledge and proficiency to perform adequate surgical techniques. Ultimately, animals may unnecessarily suffer due to morbidities from limited or poor surgical competencies. We have translated developments derived from the human surgical simulation field for application to veterinary surgical training. We present our work on intuitive software for learning regional anatomy, surgical simulations, and on several limiting factors that impede the validation and adoption of simulation technologies for use by the veterinarian surgical community. PMID- 18391356 TI - Augmented reality assistance for realization of incision planning and model-based analysis of the modified surface model. AB - In this paper an application for incision planning, AR-visualization of plan data and model-based analysis of the modified anatomical structure due to realization of the planned incision is presented. The developed algorithm detects modifications of the acquired surface model and generates a parameterized description of the wound. PMID- 18391357 TI - Medical student evaluation using virtual pathology echocardiography (VPE) for augmented standardized patients. AB - With the increasing role played by ultrasound in clinical diagnostics, ultrasound training in medical education is becoming more important. We have developed a real-time ultrasound simulator to augment standardized patients (SPs) with a simulated echocardiogram. It enables changes in the standard of ultrasound training where the clinical routine is practiced on real patients, which limits monitored and guided examinations for medical students due to time constraints and availability of patients. This paper describes our preliminary system of a tracker-based echocardiogram simulator, the latest evaluation results, and future work. PMID- 18391358 TI - Telecontrol function of an endoscopic surgical robot with two hands for tele NOTES surgery. AB - We have been developing a surgical robot system with navigation function for abdominal surgery since 2001. Clinical feasibility of the robot has been evaluated by animal experiments. Endoscopic mucosal resection (EMR) and other surgical procedures for Natural Orifice Transluminal Endoscopic Surgery (NOTES) were performed by the combined use of the robot manipulators. In this study, we developed a telecontrol system for the robot to perform an endoscopic abdominal robotic surgery using a robotic telesurgery method. By using a developed system, a telesurgery experiment was conducted between laboratories through LAN including the emulated network delay time. As a result, high-speed data transmission from the master device to the robot could be performed. We also confirmed the implementation of a telecontrol function for the surgical robot. In the near future, we will perform surgeries such as EMR and NOTES with the optimal telecontrol function. PMID- 18391359 TI - Design and simulation of a visual and haptic assisted biopsy (ViHAB) system. AB - Core needle biopsy is a non-invasive technique for confirming breast and prostate cancer. Several non-real time image based systems have been developed to guide the needle to the target. X-ray, ultrasound (US), MRI or x-ray fluoroscopy are used to guide the needle during biopsy. However, these methods are non-real time or, the imaging technique is two dimensional or, ionizing. Our broad objective is to develop a visually guided, haptically assisted breast biopsy system (ViHAB) using real time 3D US imaging and haptic guidance. ViHAB will help the radiologist identify suspicious tissue and provide real time guidance for core needle biopsy. The ViHAB simulator developed at the Virtual Environment Laboratory is capable of reading and displaying 3-D US images, keep track of micro-calcification in near real time in a sequence of images as well as provide haptic and visual guidance to a virtual needle via a joystick. PMID- 18391360 TI - Investigation into the effectiveness of vibrotactile feedback to improve the haptic realism of an arthroscopy training simulator. AB - A simulator with a custom developed haptic device named OrthoForce was developed at Imperial College for knee arthroscopy training. It is capable providing limited force bandwidth, which has shown to be suitable for the reproduction of "moderately high collisions". In recent published works, it was found that high frequency vibrations (otherwise known as vibrotactile feedback), which occur when two objects come into contact (e.g. during tapping, stroking or puncturing) can be discriminated by the human tactile system. It is hypothesised that simulating vibrotactile feedback will improve the haptic realism of hard collisions. Thus, this paper presents the development of a hardware extension to the OrthoForce system, which is able to produce timed high frequency vibrations, and its effect on the simulated haptic realism as experienced by different user groups. Although limited in scope, preliminary results are encouraging. PMID- 18391361 TI - The visible ear surgery simulator. AB - This paper presents a real-time computer simulation of surgical procedures in the ear, in which a surgeon drills into the temporal bone to gain access to the middle or inner ear. The purpose of this simulator is to support development of anatomical insight and training of drilling skills for both medical students and experienced otologists. The key contributions in this application are the visualization and interaction models in the context of ear surgical simulation. The visualization is based on an existing data set, "The Visible Ear", containing a unique volume depicting the inner ear in natural colours. The applied visualization is based on GPU ray casting, allowing high quality and flexible volume rendering using modern graphics card. In connection with the visualization model, different methods for optimizing the GPU ray casting procedure are presented, along with a method for combining polygon based graphics with volume rendering. In addition, different light models are presented that contribute to a realistic rendering of the different parts of the inner ear. To achieve a physically plausible drilling experience, a Phantom Omni force feedback device is utilized. The applied interaction model facilitates a realistic user experience of the response forces from the drilling tool. PMID- 18391362 TI - Integration of genomic and medical data into a 3D atlas of human anatomy. AB - We have developed a framework for the visual integration and exploration of multi scale biomedical data, which includes anatomical and molecular components. We have also created a Java-based software system that integrates molecular information, such as gene expression data, into a three-dimensional digital atlas of the male adult human anatomy. Our atlas is structured according to the Terminologia Anatomica. The underlying data-indexing mechanism uses open standards and semantic ontology-processing tools to establish the associations between heterogeneous data types. The software system makes an extensive use of virtual reality visualization. PMID- 18391363 TI - Image registration assists novice operators in ultrasound assessment of abdominal trauma. AB - Transcutaneous ultrasound imaging may be used to detect abdominal hemorrhage in the field setting. The Focused Assessment with Sonography for Trauma (FAST) examination was developed to characterize blunt abdominal trauma and has been shown to be effective for assessing penetrating trauma as well. However, it is unlikely that a minimally trained operator could perform a diagnostic examination. In our system, the operator is be supported by real-time 3D volume displays. The operator will be directed through the examination by prompts from a computer system or outside expert, potentially with knowledge of the anatomy of the injured patient. The key elements of the tele-operated FAST exam capability have been demonstrated; the exam is performed with real-time guidance from anatomic images registered to the body. It appears likely that Image Registration will assist hemorrhage detection at the point of injury or in the initial evaluation by a trauma response team. PMID- 18391364 TI - The use of stereolithographic hand held models for evaluation of congenital anomalies of the great arteries. AB - Imaging anomalies of the great vessels has traditionally been accomplished using conventional biplane modalities as well as three-dimensional (3D) video displays. Our aim was to review the use of stereolithography to create 3D models to assess coarctation of the aorta and vascular rings. Twelve patients had high-resolution CT scans to evaluate anomalies of the great arteries (coarctation: 9, vascular ring: 3). Ages were 19 days to 29 years and weights were 3.3 to 139 kg. Digital dicom data from each scan were converted by a commercially available software package into a 3D digital image. The area of interest was selected and the image was exported to a 3D stereolithographic printer to create a 3D model. The models were then evaluated and the results compared to catheterization and surgical findings. All models accurately displayed the pathology investigated. All 3 of the vascular ring models correlated with surgical findings (double arch: 2, pulmonary sling: 1). Models of aortic coarctation allowed clear depictions of discrete narrowing as well as arch hypoplasia and tortuosity. Stereolithography can create realistic 3D models that accurately display aortic pathology and add important additional information, which may have implications regarding surgical and transcatheter interventions and may also be useful teaching tools for parents and students. PMID- 18391365 TI - Parameter optimization for 3D mass-spring-damper models. AB - In this paper, we investigate the physical accuracy of the 3D mass-spring-damper (MSD) model of an isotropic object. The isotropic object should have the same Poisson constant and Young's modulus in different directions, and so should its model. Based on these two properties, we derive a set of constraints on the parameters of the 3D MSD model. From these constraints, the parameters of the MSD model can be obtained by the constrained least square method. For the MSD model with tetrahedral meshes, we show that its tensile stiffness can be achieved very accurately, and the prone irregular Poisson effects can be suppressed below a tolerable level although its Poisson constant generally cannot be precisely achieved. For the MSD model with hexahedral meshes, we find that the parameters of the model can be obtained explicitly in terms of material properties and mesh geometry. In this case, we also demonstrate that both the tensile stiffness and the Poisson constant can be accurately achieved. PMID- 18391366 TI - An evaluation of prototype VR medical training environment: applied surgical anatomy training for malignant breast disease. AB - This paper presents an enquiry into the suitability of Virtual Reality (VR) technology as the principal training method for applied surgical anatomy. In this work we present the development of a prototype VR medical training environment and the evaluation results of preliminary trials aiming to identify the effectiveness of the system in the subject domains of anatomy teaching and surgical rehearsal, whilst acknowledging current training requirements. PMID- 18391367 TI - Combat related post traumatic stress disorder: a multiple case report using virtual reality graded exposure therapy with physiological monitoring. AB - The current report summarizes case studies from an Office of Naval Research (ONR) funded project to compare the effects of Virtual Reality Graded Exposure Therapy (VRGET) with a treatment as usual control condition in active-duty Navy Corpsmen, Navy SeaBees and Navy and Marine Corps Support Personnel. Details of the collaborative program between the Virtual Reality Medical Center (VRMC) and Naval Medical Center San Diego (NMCSD) will be discussed. PMID- 18391368 TI - Dental fiberscope with navigation system for endodontic treatments. AB - We developed a dental fiberscope with a navigation system. The aim of this study is to direct the position of the tip of the endoscope fiber in the root canal via our navigation system. The distinguishing characteristic of our system is that it could be particularly practical in narrow spaces, such as those involving endodontic treatment typical of root canals. We were subsequently able to precisely indicate the relation of the position between the device and the teeth on the 3D model on the monitor. We inspected our navigation system using both the phantom model and the reconstructed 3D model. From this we could comprehend the relation of the position between the teeth and the device, and aim precisely at the lesion. If we are able to realize the correct position of the endoscope, we can safely and accurately deliver laser irradiation to the lesion. Consequently, the application of the endoscopic navigation system could increase the success rate for root canal treatments with recalcitrant lesions. PMID- 18391369 TI - Improvement of colonoscopy skills through simulation-based training. AB - The objective of this study is to determine whether targeted colonoscopy skills are acquired through simulation-based training using the KAIST-Ewha Colonoscopy Simulator II, and the acquired skills can be transferred to colonoscopy to actual patients. Eleven subjects consisting of six fellows and five residents participated in the study. The fellows and residents were divided into two groups, simulation-trained group and control group. Simulation-based training included practicing the targeted skills of colonoscopy using two training scenarios with different colon flexures and degrees of difficulty. The trainees were requested to practice until they reach all the established training goals. The both groups were evaluated during their colonoscopies to actual patients, which were performed under close supervision of colonoscopy experts. The results of this study show that the developed colonoscopy simulator is effective in teaching the targeted colonoscopy skills, and transferring those skills to actual colonoscopy. PMID- 18391370 TI - Hexacopper(I) phosphorus(V) bromide penta(selenide/sulfide), Cu6P(Se0.7S0.3)5Br. AB - This work illustrates possible diffusion paths for Cu(I) ions in a highly disordered structure of a superionic conductor of the argyrodite family. The Cu(6)P(Se(0.7)S(0.3))(5)Br cubic structure is built from a [P(Se(0.7)S(0.3))(5)Br] framework in which Cu(I) ions are distributed in various tetrahedral, triangular and linear sites. There are two types of disorder in the structure. The first type results from the fact that there are fewer Cu(I) ions than the number of positions available for them in the unit cell. The second type is due to the static distribution of Se and S atoms in the [P(Se(0.7)S(0.3))(5)Br] framework. The title compound is a solid solution of two efficient ionic conductors, namely Cu(6)PSe(5)Br and Cu(6)PS(5)Br, in which high ionic conductivity results from order-disorder phenomena in the copper substructure. To shed light on the distribution of Cu(I) ions in disordered Cu(6)P(Se(0.7)S(0.3))(5)Br, we refined their positions using a combination of a nonharmonic approach and a split-atom model. At room temperature, Cu(I) ions show strong anharmonic vibrations along the edge of the (Br)(4) tetrahedra. The probability density functions of the Cu(I) ions overlap and reveal possible diffusion paths. PMID- 18391371 TI - K2[O(HgSO3)3], a new sulfitomercurate with an [OHg3] core. AB - The structure of dipotassium mu(3)-oxido-tris[sulfitomercurate(II)], K(2)[O(HgSO(3))(3)], is characterized by segregation of the K(+) cations and complex [O(HgSO(3))(3)](2-) anions into layers parallel to (010). The anion has m symmetry and is a new example of a mu(3)-oxido-trimercurate complex with a central [OHg(3)] core. This unit adopts the shape of a flat, almost trigonal, pyramid (mean O-Hg = 2.072 A and mean Hg-O-Hg = 110.8 degrees). The two independent Hg-S bonds have nearly the same length (mean Hg-S = 2.335 A). Due to intermolecular O...Hg donor-acceptor interactions greater than 2.65 A, the O-Hg-S fragments are slightly bent. The [KO(9)] coordination polyhedron of the K(+) cation approaches a distorted tricapped trigonal prism with a [6+1+2] coordination. PMID- 18391372 TI - Ba3Li2V2O7Cl4, a new vanadate with a channel structure. AB - The tribarium dilithium divanadate tetrachloride Ba(3)Li(2)V(2)O(7)Cl(4) is a new vanadate with a channel structure and the first known vanadate containing both Ba and Li atoms. The structure contains four non-equivalent Ba(2+) sites (two with m and two with 2/m site symmetry), two Li(+) sites, two nonmagnetic V(5+) sites, five O(2-) sites (three with m site symmetry) and four Cl(-) sites (m site symmetry). One type of Li atom lies in LiO(4) tetrahedra (m site symmetry) and shares corners with VO(4) tetrahedra to form eight-tetrahedron Li(3)V(5)O(24) rings and six-tetrahedron Li(2)V(4)O(18) rings; these rings are linked within porous layers parallel to the ab plane and contain Ba(2+) and Cl(-) ions. The other Li atoms are located on inversion centres and form isolated chains of face sharing LiCl(6) octahedra. PMID- 18391373 TI - 1,5-Asymmetric induction during nucleophilic additions to arenetricarbonylchromium complexes: tricarbonyl(eta(6)-1-methyl-4-{spiro[(1R,2S) 1,7,7-trimethylbicyclo[2.2.1]heptane-3,2'-1,3-dioxolan]-2-yloxy}benzene)chromium. AB - The tricarbonylchromium unit bound to the arene ring of the chiral title complex, [Cr(C(19)H(26)O(3))(CO)(3)], is rotated by ca 25 degrees in agreement with the proposed mechanism for 1,5-asymmetric induction of nucleophilic attack. PMID- 18391374 TI - Diacetonitrile-1kappaN,3kappaN-bis{mu-trans-N-[3-(dimethylamino)propyl]-N'-(2 hydroxyethyl)oxamidato(2-)}-1:2kappa(5)N,N',O:O',N'';2:3kappa(5)O',N'':N,N',O dithiocyanato-1kappaN,3kappaN-tricopper(II). AB - The title compound, [Cu(3)(C(9)H(17)N(3)O(3))(2)(NCS)(2)(CH(3)CN)(2)], contains two square-pyramidal Cu(II) units chelated by a transoid asymmetrical N-[3 (dimethylamino)propyl]-N'-(2-hydroxyethyl)oxamidate (dmapheoxd) dianion {H(2)dmapheoxd is N-[3-(dimethylamino)propyl]-N'-(2-hydroxyethyl)oxamide}, which coordinates to another Cu(II) ion in a square-planar environment lying on a crystallographic inversion center. Thus, the trans-oxamide ligand bridges two Cu(II) ions with different coordination numbers, and this is the first instance of such a zero-dimensional oxamide-bridged complex. The activated methyl group in the coordinated acetonitrile molecule is involved in a strong nonclassical C H...O hydrogen bond, which contributes to a one-dimensional chain extending in the b direction. Considering the presence of weak bonding between the Cu atom and the uncoordinated hydroxyl O atoms, a two-dimensional structure is formed parallel to the ab plane. PMID- 18391375 TI - Dibromido(dimethyl sulfoxide-kappaO)(1,10-phenanthroline-kappa(2)N,N')copper(II). AB - The solvent effect on the molecular structures of copper(II) complexes produced from the reaction between CuBr(2) and 1,10-phenanthroline is evident. The monomeric title compound, [CuBr(2)(C(12)H(8)N(2))(C(2)H(6)OS)], which consists of discrete units, is produced from this reaction in dimethyl sulfoxide (DMSO), whereas a polymeric copper(II) compound is known to be produced from the same reaction in the poor coordinating solvent ethanol. The geometry around the copper(II) ion in the title compound is best described as trigonal-bipyramidal distorted square-based pyramidal, with a tau value of 0.37. The two phenanthroline N atoms, the DMSO O atom and one of the Br atoms occupy the four basal positions, while the second Br atom occupies the axial position. The magnetic susceptibility data also indicate that the title compound is monomeric, but there is still a weak antiferromagnetic interaction between paramagnetic copper(II) centers via the intermolecular 'Cu-Br...Br-Cu' contact pathway. PMID- 18391376 TI - Mononuclear nickel(II) and zinc(II) complexes with deprotonated forms of the tripodal hexadentate ligand 1,3-bis(2-hydroxybenzylidene)-2-(2 hydroxybenzylideneaminomethyl)-2-methylpropane-1,3-diamine. AB - In the crystal structures of both title compounds, [1,3-bis(2-hydroxybenzylidene) 2-methyl-2-(2-oxidobenzylideneaminomethyl)propane-1,3-diamine]nickel(II) [2-(2 hydroxybenzylideneaminomethyl)-2-methyl-1,3-bis(2-oxidobenzylidene)propane-1,3 diamine]nickel(II) chloride methanol disolvate, [Ni(C(26)H(25.5)N(3)O(3))](2)Cl x 2 CH(4)O, and [1,3-bis(2-hydroxybenzylidene)-2-methyl-2-(2 oxidobenzylideneaminomethyl)propane-1,3-diamine]zinc(II) perchlorate [2-(2 hydroxybenzylideneaminomethyl)-2-methyl-1,3-bis(2-oxidobenzylidene)propane-1,3 diamine]zinc(II) methanol trisolvate, [Zn(C(26)H(25)N(3)O(3))]ClO(4) x [Zn(C(26)H(26)N(3)O(3))] x 3 CH(4)O, the 3d metal ion is in an approximately octahedral environment composed of three facially coordinated imine N atoms and three phenol O atoms. The two mononuclear units are linked by three phenol phenolate O-H...O hydrogen bonds to form a dimeric structure. In the Ni compound, the asymmetric unit consists of one mononuclear unit, one-half of a chloride anion and a methanol solvent molecule. In the O-H...O hydrogen bonds, two H atoms are located near the centre of O...O and one H atom is disordered over two positions. The Ni(II) compound is thus formulated as [Ni(H(1.5)L)](2)Cl x 2 CH(3)OH [H(3)L is 1,3-bis(2-hydroxybenzylidene)-2-(2 hydroxybenzylideneaminomethyl)-2-methylpropane-1,3-diamine]. In the analogous Zn(II) compound, the asymmetric unit consists of two crystallographically independent mononuclear units, one perchlorate anion and three methanol solvent molecules. The mode of hydrogen bonding connecting the two mononuclear units is slightly different, and the formula can be written as [Zn(H(2)L)]ClO(4) x [Zn(HL)] x 3 CH(3)OH. In both compounds, each mononuclear unit is chiral with either a Delta or a Lambda configuration because of the screw coordination arrangement of the achiral tripodal ligand around the 3d metal ion. In the dimeric structure, molecules with Delta-Delta and Lambda-Lambda pairs co-exist in the crystal structure to form a racemic crystal. A notable difference is observed between the M-O(phenol) and M-O(phenolate) bond lengths, the former being longer than the latter. In addition, as the ionic radius of the metal ion decreases, the M-O and M-N bond distances decrease. PMID- 18391377 TI - Low-dimensional compounds containing cyano groups. XVI. (Dicyanamido kappaN1)bis(1,10-phenanthroline-kappa(2)N,N')copper(II) tetrafluoridoborate. AB - The title compound, [Cu{N(CN)(2)}(C(12)H(8)N(2))(2)]BF(4), was prepared as part of our study of the shape of coordination polyhedra in five-coordinated copper(II) complexes. Single-crystal X-ray analysis reveals that the structure consists of [Cu{N(CN)(2)}(phen)(2)](+) cations (phen is 1,10-phenanthroline) and BF(4)(-) anions. The Cu centre is five-coordinated in a distorted trigonal bipyramidal manner by four N atoms of two phen ligands and one N atom of a dicyanamide anion, which is coordinated in the equatorial plane at a distance of 1.996 (2) A. The two axial Cu-N(phen) distances have similar values [average 1.994 (6) A] and are shorter than the two equatorial Cu-N(phen) bonds [average 2.09 (6) A]. This work demonstrates the effect of ligand rigidity on the shape of coordination polyhedra in five-coordinated copper(II) complexes. PMID- 18391378 TI - catena-Poly[[(acetato-kappa(2)O,O')triaquabarium(II)]-mu3-acetato kappa(4)O:O,O':O']. AB - The present form of barium acetate, formulated as [Ba(C(2)H(3)O(2))(2)(H(2)O)(3)](n), is the largest reported hydrate of the salt and this leads to a distinct structural behaviour setting it apart from the rest of the family. The compound is a linear polymer with a nine-coordinate Ba(O(aqua))(3)(O(acetate))(6) monomer unit. The non-H part of the structure is ordered according to C2/m symmetry, while the disordered water H atoms only abide by this symmetry in a statistical sense. Each molecule is halved by a mirror plane bisecting the Ba centre, one water molecule and one acetate ligand, while containing the other acetate ligand. The chains are interconnected by a disordered water-water/acetate O-H...O hydrogen-bonding network involving all water H atoms. The structure and stability of this phase are compared with the other known acetates of barium which differ in the degree of hydration. PMID- 18391379 TI - A mixed bridged trinuclear copper(II)-1,10-phenanthroline complex with a linear structure. AB - The title compound, diaqua-1kappaO,3kappaO-di-mu-hydroxido 1:2kappa(2)O:O,2:3kappa(2)O:O-di-mu-methacrylato-1:2kappa(2)O:O',2:3kappa(2)O:O' bis(1,10-phenanthroline)-1kappa(2)N,N';3kappa(2)N,N'-tricopper(II) dinitrate dihydrate, [Cu(3)(C(4)H(5)O(2))(2)(OH)(2)(C(12)H(8)N(2))(2)(H(2)O)(2)](NO(3))(2) x 2 H(2)O, has the central Cu atom on an inversion centre. The three Cu(II) atoms are in a linear arrangement linked by methacrylate and hydroxide groups. The coordination environments of the Cu(II) ions are five-coordinated distorted square-pyramidal for the outer Cu atoms and four-coordinated square-planar for the central Cu atom. All nitrate ions, hydroxide groups and water molecules are linked by hydrogen bonds, forming a linear structure. The complex exhibits ferromagnetic exchange coupling, which is helpful in elucidating magnetic interactions between copper ions and other metallic ions in heteronuclear complexes. PMID- 18391380 TI - Poly[(mu2-2,2-dimethylpropane-1,3-diyl diisocyanide)-mu2-nitrato-silver(I)]: a powder study. AB - In order to investigate the effect of counter-anions on the polymeric structure of (2,2-dimethylpropane-1,3-diyl diisocyanide)silver(I) complexes, the novel title polymeric compound, [Ag(NO(3))(C(7)H(10)N(2))](n), has been synthesized. The crystal structure was determined by simulated annealing from X-ray powder diffraction data collected at room temperature. The current structure is similar to the recently reported structure of the analogue with chloride replacing nitrate. This study illustrates that both the chloride and nitrate complexes crystallize in the orthorhombic system in the Pbca space group with one monomer in the asymmetric unit, and also gives a strong indication that the counter-anion does not have a considerable effect on the polymeric structure of the complex. The Ag centre lies in a distorted tetrahedral environment and is bonded to two 2,2-dimethylpropane-1,3-diyl diisocyanide ligands to form chains. The nitrate anions crosslink the Ag centres of the chains to form a two-dimensional polymeric network structure. PMID- 18391381 TI - Poly[[diaquabis(mu2-1-oxo-2,6,7-trioxa-1-phosphabicyclo[2.2.2]octane-4 carboxylato)copper(II)] dihydrate]. AB - The asymmetric unit of the title complex, {[Cu(C(5)H(6)O(6)P)(2)(H(2)O)(2)] x 2 H(2)O}(n), consists of half a Cu atom, one complete 1-oxo-2,6,7-trioxa-1 phosphabicyclo[2.2.2]octane-4-carboxylate anion ligand and two non-equivalent water molecules. The Cu atom lies on a crystallographic inversion centre and has an elongated axially distorted octahedral environment. A two-dimensional layer structure parallel to (100) is formed as a result of the connectivity brought about by each anion bonding to two different Cu atoms via a carboxylate O atom and a bridging O atom of a C-O-P group. The water molecules participate in extensive O-H...O hydrogen bonding. Neighbouring layers are linked together by intermolecular hydrogen-bonding interactions. The crystal structure is characterized by intra- and interlayer motifs of a hydrogen-bonded network. This study demonstrates the usefulness of carboxylates with caged phosphate esters in crystal engineering. PMID- 18391382 TI - Hexaquacobalt(II) bis(5-hydroxy-7-methoxy-4-oxo-2-phenyl-4H-chromene-6-sulfonate) tetrahydrate. AB - The title compound, [Co(H(2)O)(6)](C(16)H(11)O(7)S)(2) x 4 H(2)O, with cobalt(II) at the centre of symmetry, exhibits alternating hydrophilic and hydrophobic regions. Hydrophilic regions are generated by O-H...O hydrogen bonds among sulfonate groups, involving solvent water molecules and coordinated water molecules; pi-pi stacking interactions assemble the flavone skeletons into columns which form the hydrophobic regions. A three-dimensional network is built up from an extensive array of hydrogen bonds, pi-pi stacking interactions and electrostatic interactions between the cation and anion. As a salt of the sulfonated derivative of naturally occurring tectochrysin (5-hydroxy-7 methoxyflavone), this compound offers enhanced solubility and potential biological activity over the natural product. PMID- 18391383 TI - (N-{[4-(1,3-benzothiazol-2-yl)anilino]carbonylmethyl-kappaO}iminodiacetato kappa(3)O,N,O')(1,10-phenanthroline-kappa(2)N,N')cobalt(II) pentahydrate. AB - The title compound, [Co(C(19)H(15)N(3)O(5)S)(C(12)H(8)N(2))] x 5 H(2)O, has a moderately distorted octahedral coordination environment composed of two N atoms of a 1,10-phenanthroline ligand and one N and three O atoms of an N-{[4-(1,3 benzothiazol-2-yl)anilino]carbonylmethyl}iminodiacetate (ZL-5(2-)) ligand. The ring systems of the phenanthroline and ZL-5(2-) ligands are coplanar and the complexes pack in layers parallel to the ab plane with the rings of adjacent complexes facing one another. The layers stack along the c axis and are linked by hydrogen bonds involving the five water solvent molecules in the asymmetric unit and O atoms of the acetate groups of the ZL-5(2-) ligand. This is believed to be the first crystal structure of a complex of a 2-(4-aminophenyl)benzothiazole ligand. PMID- 18391384 TI - Bis{mu-(E)-2-[(2-pyridylamino)(2-pyridylimino)methyl]benzato}bis[acetatozinc(II)] tetrahydrate. AB - In the C(2)-symmetric dinuclear title complex, [Zn(2)(C(18)H(13)N(4)O(2))(2)(C(2)H(3)O(2))(2)] x 4 H(2)O, each Zn(II) ion is five-coordinated in a distorted trigonal bipyramidal fashion by one carboxylate O atom from one benzoate ligand, one imine N atom and two pyridyl N atoms from a second benzoate ligand, and one O atom from an acetate anion. The two Zn atoms are bridged by the two benzoate ligands, forming a dinuclear structure with a 14 membered macrocycle. Adjacent dinuclear units are further connected by extensive hydrogen bonds involving the solvent water molecules, giving a three-dimensional hydrogen-bonded framework. The framework can be regarded as an example of the four-connected node network of the PtS topology. PMID- 18391385 TI - 5-Iodobenzofurazan 1-oxide: polymorphs, pseudosymmetry and disorder. AB - 5-Iodobenzofurazan 1-oxide (systematic name: 5-iodobenzo-1,2,5-oxadiazole 1 oxide), C(6)H(3)IN(2)O(2), occurs in two polymorphic forms, both monoclinic in P2(1)/c with Z' = 2. The intermolecular interactions in the two polymorphs are quite different. In polymorph (I), there are strong intermolecular I...O interactions, with I...O distances of 3.114 (8) and 3.045 (8) A. In polymorph (II), there are strong intermolecular I...N interactions, with I...N distances of 3.163 (4) and 3.175 (5) A. In (I), there is about 15% disorder in one molecule and about 5% in the other. In both polymorphs, there are pseudosymmetric relationships between the crystallographically independent molecules. PMID- 18391386 TI - Hydrogen-bonded chains of rings in 1-(4-chloroanilinomethyl)-5-(4-chlorophenyl) 1,3,5-triazinane-2-thione and hydrogen-bonded sheets in 1-anilinomethyl-5-phenyl 1,3,5-triazinane-2-thione. AB - In 1-(4-chloroanilinomethyl)-5-(4-chlorophenyl)-1,3,5-triazinane-2-thione, C(16)H(16)Cl(2)N(4)S, there are two independent molecules in the asymmetric unit which form inversion dimers via two weak N-H...S hydrogen bonds. The dimers are then linked into C(9)C(14) chains by a C-H...S hydrogen bond and a C-H...Cl contact. In 1-(anilinomethyl)-5-phenyl-1,3,5-triazinane-2-thione, C(16)H(18)N(4)S, molecules are linked into complex sheets via a combination of N H...S and C-H...pi hydrogen bonds. PMID- 18391387 TI - Geometry and bond-length alternation in nonlinear optical materials. II. Effects of donor strength in two push-pull molecules. AB - The compounds N-[2-(4-cyano-5-dicyanomethylene-2,2-dimethyl-2,5-dihydrofuran-3 yl)vinyl]-N-phenylacetamide, C(20)H(16)N(4)O(2), (I), and 2-{3-cyano-5,5-dimethyl 4-[2-(piperidin-1-yl)vinyl]-2,5-dihydrofuran-2-ylidene}malononitrile 0.376 hydrate, C(17)H(18)N(4)O x 0.376 H(2)O, (II), are novel push-pull molecules. The significant bonding changes in the polyene chain compared with the parent molecule 2-dicyanomethylene-4,5,5-trimethyl-2,5-dihyrofuran-3-carbonitrile are consistent with the relative electron-donating properties of the acetanilido and piperidine groups. The packing of (I) utilizes one phenyl-cyano C-H...N and two phenyl-carbonyl C-H...O hydrogen bonds. Compound (II) crystallizes with a partial water molecule (0.376H(2)O), consistent with cell packing that is dominated by attractive C-H...N(cyano) interactions. These compounds are precursors to novel nonlinear optical chromophores, studied to assess the impact of donor strength and the extent of conjugation on bond-length alternation, crystal packing and aggregation. PMID- 18391388 TI - Five bicyclo[3.3.0]octa-2,6-dienes. AB - A series of five compounds containing the bicyclo[3.3.0]octa-2,6-diene skeleton are described, namely tetramethyl cis,cis-3,7-dihydroxybicyclo[3.3.0]octa-2,6 diene-2,4-exo,6,8-exo-tetracarboxylate, C(16)H(18)O(10), (I), tetramethyl cis,cis 3,7-dihydroxy-1,5-dimethylbicyclo[3.3.0]octa-2,6-diene-2,4-exo,6,8-exo tetracarboxylate, C(18)H(22)O(10), (II), tetramethyl cis,cis-3,7 dimethoxybicyclo[3.3.0]octa-2,6-diene-2,4-exo,6,8-exo-tetracarboxylate, C(18)H(22)O(10), (III), tetramethyl cis,cis-3,7-dimethoxy-1,5 dimethylbicyclo[3.3.0]octa-2,6-diene-2,4-exo,6,8-exo-tetracarboxylate, C(20)H(26)O(10), (IV), and tetramethyl cis,cis-3,7-diacetoxybicyclo[3.3.0]octa 2,6-diene-2,4-exo,6,8-exo-tetracarboxylate, C(20)H(22)O(12), (V). The bicyclic core is substituted in all cases at positions 2, 4, 6 and 8 with methoxycarbonyl groups and additionally at positions 3 and 7 with hydroxy [in (I) and (II)], methoxy [in (III) and (IV)] or acetoxy [in (V)] groups. The conformations of the methoxycarbonyl groups at positions 2 and 4 are exo for all five compounds. Each C(5) ring of the bicyclic skeleton is almost planar, but the rings are not coplanar, with dihedral angles of 54.93 (7), 69.85 (5), 64.07 (4), 80.74 (5) and 66.91 (7) degrees for (I)-(V), respectively, and the bicyclooctadiene system adopts a butterfly-like conformation. Strong intramolecular hydrogen bonds exist between the -OH and C=O groups in (I) and (II), with O...O distances of 2.660 (2) and 2.672 (2) A in (I), and 2.653 (2) and 2.635 (2) A in (II). The molecular packing is stabilized by weaker C-H...O(=C) interactions, leading to dimers in (I)-(III) and to a chain structure in (V). The structure series presented in this article shows how the geometry of the cycloocta-2,6-diene skeleton changes upon substitution in different positions and, consequently, how the packing is modified, although the intermolecular interactions are basically the same across the series. PMID- 18391389 TI - Delta-sulfanilamide. AB - The delta polymorph of sulfanilamide (or 4-aminobenzenesulfonamide), C(6)H(8)N(2)O(2)S, displays an overall three-dimensional hydrogen-bonded network that is dominated by a two-dimensional substructure with R(2)(2)(8) rings; these result from dimeric N-H...O interactions between adjacent sulfonamide groups. This study shows how the polymorphism of sulfanilamide is linked to its versatile hydrogen-bonding capabilities. PMID- 18391390 TI - Conformation of hydrogen-bonded dimeric o-methyl-substituted benzoic acids. AB - Molecules of 2,4-dimethylbenzoic acid, C(9)H(10)O(2), form typical centrosymmetric hydrogen-bonded dimers. The carboxyl group is twisted with respect to the benzene ring and the methyl group in the ortho position shows evasive in-plane splaying. The relation between the in-plane splaying and the twist angle of the carboxyl group for various ortho-substituted dimeric derivatives of benzoic acid is presented. It shows how the steric strains are released depending on the numbers and positions of the substituents. PMID- 18391391 TI - 2,6-Dimethoxy-7,9-dimethylpurinium iodide hemihydrate. AB - In the title compound, C(9)H(13)N(4)O(2)(+) x I(-) x 0.5 H(2)O, the non-H atoms of the ionic components lie on a mirror plane in Cmca, with the O atom of the partial water molecule lying on a twofold rotation axis. Whereas one of the methoxy methyl groups is directed away from the adjacent N-methyl group, the other methoxy methyl group is directed towards its adjacent N-methyl group. The conformation of the methoxy methyl groups provides an explanation for the outcomes of intramolecular thermal rearrangements of 2,6-dialkoxy-7,9 dimethylpurinium salts. PMID- 18391392 TI - Accurate stereochemistry for two related 22,26-epiminocholestene derivatives. AB - Regioselective opening of ring E of solasodine under various conditions afforded (25R)-22,26-epiminocholesta-5,22(N)-diene-3beta,16beta-diyl diacetate (previously known as 3,16-diacetyl pseudosolasodine B), C(31)H(47)NO(4), or (22S,25R)-16beta hydroxy-22,26-epiminocholesta-5-en-3beta-yl acetate (a derivative of the naturally occurring alkaloid oblonginine), C(29)H(47)NO(3). In both cases, the reactions are carried out with retention of chirality at the C16, C20 and C25 stereogenic centers, which are found to be S, S and R, respectively. Although pseudosolasodine was synthesized 50 years ago, these accurate assignments clarify some controversial points about the actual stereochemistry for these alkaloids. This is of particular importance in the case of oblonginine, since this compound is currently under consideration for the treatment of aphasia arising from apoplexy; the present study defines a diastereoisomerically pure compound for pharmacological studies. PMID- 18391393 TI - 1H-indazole and 2H-indazole derivatives of androsta-5,16-dien-3beta-ol. AB - The title compounds, 17-(1H-indazol-1-yl)androsta-5,16-dien-3beta-ol, (I), and 17 (2H-indazol-2-yl)androsta-5,16-dien-3beta-ol, (II), both C(26)H(32)N(2)O, have an indazole substituent at the C17 position. The six-membered B ring of each compound assumes a half-chair conformation. A twist of the steroid skeleton is observed and reproduced in quantum-mechanical ab initio calculations of the isolated molecule using a molecular orbital Hartree-Fock method. In the 1H indazole derivative, (I), the molecules are joined in a head-to-head fashion via O-H...O hydrogen bonds, forming chains along the a axis. In the 2H-indazole derivative, (II), the molecules are joined in a head-to-tail fashion with one of the N atoms of the indazole ring system acting as the acceptor. The hydrogen-bond pattern consists of zigzag chains running along the b axis. Substituted steroids have proven to be effective in inhibiting androgen biosynthesis through coordination of the Fe atoms of some enzymes, and this study shows that indazole substituted steroids adopt twisted conformations that restrict their intermolecular interactions. PMID- 18391394 TI - Hydrogen-bonded sheet structures in neutral, anionic and hydrated 6-amino-2 (morpholin-4-yl)-5-nitrosopyrimidines. AB - In each of 6-amino-3-methyl-2-(morpholin-4-yl)-5-nitrosopyrimidin-4(3H)-one, C(9)H(13)N(5)O(3), (I), morpholin-4-ium 4-amino-2-(morpholin-4-yl)-5-nitroso-6 oxo-1,6-dihydropyrimidin-1-ide, C(4)H(10)NO(+) x C(8)H(10)N(5)O(3)(-), (II), and 6-amino-2-(morpholin-4-yl)-5-nitrosopyrimidin-4(3H)-one hemihydrate, C(8)H(11)N(5)O(3) x 0.5 H(2)O, (III), the bond distances within the pyrimidine components are consistent with significant electronic polarization, which is most marked in (II) and least marked in (I). Despite the high level of substitution, the pyrimidine rings are all effectively planar, and in each of the pyrimidine components, there are intramolecular N-H...O hydrogen bonds. In each compound, the organic components are linked by multiple N-H...O hydrogen bonds to form sheets of widely differing construction, and in compound (III) adjacent sheets are linked by the water molecules, so forming a three-dimensional hydrogen-bonded framework. This study also contains the first direct geometric comparison between the electronic polarization in a neutral aminonitrosopyrimidine and that in its ring-deprotonated conjugate anion in a metal-free environment. PMID- 18391395 TI - Hydrogen bonding in the bromide salts of 4-aminobenzoic acid and 4 aminoacetophenone. AB - In the title compounds, 4-carboxyanilinium bromide, C(7)H(8)NO(2)(+) x Br(-), (I), and 4-acetylanilinium bromide, C(8)H(10)NO(+) x Br(-), (II), each asymmetric unit contains a discrete cation with a protonated amino group and a halide anion. Both crystal structures are characterized by two-dimensional hydrogen-bonded networks. The ions in (I) are connected via N-H...Br, N-H...O and O-H...Br hydrogen bonds, with three characteristic graph-set motifs, viz. C(8), C(2)(1)(4) and R(3)(2)(8). The centrosymmetric hydrogen-bonded R(2)(2)(8) dimer motif characteristic of carboxylic acids is absent. The ions in (II) are connected via N-H...Br and N-H...O hydrogen bonds, with two characteristic graph-set motifs, viz. C(8) and R(4)(2)(8). The significance of this study lies in its illustration of the differences between the supramolecular aggregations in two similar compounds. The presence of the methyl group in (II) at the site corresponding to the hydroxyl group in (I) results in a significantly different hydrogen-bonding arrangement. PMID- 18391396 TI - (1-Pyridinio)perfluorophenacylide: a new stable pyridinium ylide in the enol form. AB - The title compound, C(13)H(6)F(5)NO, exists in the enol form and adopts the E configuration about the enol double bond. It is the first example of an enol-type pyridinium ylide. The enol structure was unambiguously determined on the basis of the significantly longer C-O bond and shorter C-C bond. Intramolecular C-H...O and C-H...F hydrogen bonds are responsible for promotion of the enol form and for the stability of this compound. PMID- 18391397 TI - Ethyl 2-methoxy-6-[(triphenylphosphoranylidene)amino]nicotinate and ethyl 2 methylsulfanyl-6-[(triphenylphosphoranylidene)amino]nicotinate. AB - The molecules of ethyl 2-methoxy-6-[(triphenylphosphoranylidene)amino]nicotinate, C(27)H(25)N(2)O(3)P, (I), and ethyl 2-methylsulfanyl-6 [(triphenylphosphoranylidene)amino]nicotinate, C(27)H(25)N(2)O(2)PS, (II), have almost identical bond lengths and molecular conformations, and both show evidence for polarized electronic structures. However, the crystal structures, as illustrated by the weak hydrogen bonds linking the molecules, are significantly different. The significance of this study lies in the observation that two compounds which are almost identical in constitution, configuration and conformation nonetheless adopt different crystal structures. PMID- 18391398 TI - Hydrate frameworks involving the pyridazino[4,5-d]pyridazine unit as a multiple hydrogen-bond acceptor. AB - 1,4,5,8-Tetramethylpyridazino[4,5-d]pyridazine trihydrate, C(10)H(12)N(4) x 3 H(2)O, (I), and 1,2,3,6,7,8-hexahydrocinnolino[5,4,3-cde]cinnoline tetrahydrate, C(12)H(12)N(4) x 4 H(2)O, (II), exhibit exceptional functionality of the condensed N(4)-heteroaromatic frame as a symmetric acceptor of four hydrogen bonds [N...O = 2.843 (2)-2.8716 (10) A]. Thus, all the N atoms of the electron deficient and highly pi-acidic polynitrogen heterocycles function as lone-pair donors. In (I), all the molecular components lie on or across special positions; the site symmetry is 2/m for the organic and m2m and m for the two water molecules. In (II), the organic polycycle lies across a crystallographic inversion center. Both structures involve a hydrogen-bonded centrosymmetric water pyridazine dimer as the basic supramolecular unit, which is integrated into two dimensional [in (I)] and three-dimensional [in (II)] hydrate frameworks by hydrogen bonding with the additional water molecules [O...O = 2.744 (2)-2.8827 (19) A]. The hydrate connectivity exists in the form of an (H(2)O)(3) trimer in (I) and as a one-dimensional zigzag (H(2)O)(n) chain in (II). PMID- 18391399 TI - 2,4-Bis[1-(4-hydroxyphenyl)-1-methylethyl]phenol: a three-dimensional framework built from O-H...O hydrogen bonds. AB - In the crystal structure of the title compound, C(24)H(26)O(3), (I), there are three different O-H...O hydrogen bonds, which individually form chains of C(10), C(12) and C(16) types. The combined effect of all these hydrogen bonds is the formation of a three-dimensional network, which is additionally stabilized by a single intermolecular C-H...pi interaction. The significance of this study lies in the comparison drawn between the molecular structure of (I) and those of several of its analogues, which shows a close similarity in the almost perpendicular orientation of the benzene rings. PMID- 18391400 TI - Deposition of diffraction images to be discussed at the Open Meeting of the Commission on Biological Macromolecules of the IUCr in Osaka. PMID- 18391401 TI - Structure of wild-type Plk-1 kinase domain in complex with a selective DARPin. AB - As a key regulator of mitosis, the Ser/Thr protein polo-like kinase-1 (Plk-1) is a well validated drug target in cancer therapy. In order to enable structure guided drug design, determination of the crystal structure of the kinase domain of Plk-1 was attempted. Using a multi-parallel cloning and expression approach, a set of length variants were identified which could be expressed in large amounts from insect cells and which could be purified to high purity. However, all attempts to crystallize these constructs failed. Crystals were ultimately obtained by generating designed ankyrin-repeat proteins (DARPins) selective for Plk-1 and using them for cocrystallization. Here, the first crystal structure of the kinase domain of wild-type apo Plk-1, in complex with DARPin 3H10, is presented, underlining the power of selective DARPins as crystallization tools. The structure was refined to 2.3 A resolution and shows the active conformation of Plk-1. It broadens the basis for modelling and cocrystallization studies for drug design. The binding epitope of 3H10 is rich in arginine, glutamine and lysine residues, suggesting that the DARPin enabled crystallization by masking a surface patch which is unfavourable for crystal contact formation. Based on the packing observed in the crystal, a truncated DARPin variant was designed which showed improved binding characteristics. PMID- 18391402 TI - Towards a rational approach for heavy-atom derivative screening in protein crystallography. AB - Heavy-atom derivatization is routinely used in protein structure determination and is thus of critical importance in structural biology. In order to replace the current trial-and-error heavy-atom derivative screening with a knowledge-based rational derivative-selection method, the reactivity of more than 40 heavy-atom compounds over a wide range of buffer and pH values was systematically examined using peptides which contained a single reactive amino-acid residue. Met-, Cys- and His-containing peptides were derivatized against Hg, Au and Pt compounds, while Tyr-, Glu-, Asp-, Asn- and Gln-containing peptides were assessed against Pb compounds. A total of 1668 reactive conditions were examined using mass spectrometry and were compiled into heavy-atom reactivity tables (http://sis.niaid.nih.gov/cgi-bin/heavyatom_reactivity.cgi). The results showed that heavy-atom derivatization reactions are highly linked to buffer and pH, with the most accommodating buffer being MES at pH 6. A group of 21 compounds were identified as most successful irrespective of ligand or buffer/pH conditions. To assess the applicability of the peptide heavy-atom reactivity to proteins, lysozyme crystals were derivatized with a list of peptide-reactive compounds that included both known and new compounds for lysozyme derivatization. The results showed highly consistent heavy-atom reactivities between the peptides and lysozyme. PMID- 18391403 TI - Structural adaptation of endonuclease I from the cold-adapted and halophilic bacterium Vibrio salmonicida. AB - The crystal structure of the periplasmic/extracellular endonuclease I from Vibrio salmonicida has been solved to 1.5 A resolution and, in comparison to the corresponding endonucleases from V. cholerae and V. vulnificus, serves as a model system for the investigation of the structural determinants involved in the temperature and NaCl adaptation of this enzyme class. The overall fold of the three enzymes is essentially similar, but the V. salmonicida endonuclease displays a significantly more positive surface potential than the other two enzymes owing to the presence of ten more Lys residues. However, if the optimum salt concentrations for the V. salmonicida and V. cholerae enzymes are taken into consideration in the electrostatic surface-potential calculation, the potentials of the two enzymes become surprisingly similar. The higher number of basic residues in the V. salmonicida protein is therefore likely to be a result, at least in part, of adaptation to the more saline habitat of V. salmonicida (seawater) than V. cholerae (brackish water). The hydrophobic core of all three enzymes is almost identical, but the V. salmonicida endonuclease has a slightly lower number of internal hydrogen bonds. This, together with repulsive forces between the basic residues on the protein surface of V. salmonicida endonuclease I and differences in the distribution of salt bridges, probably results in higher flexibility of regions of the V. salmonicida protein. This is likely to influence both the catalytic activity and the stability of the protein. PMID- 18391404 TI - Structures of the N47A and E109Q mutant proteins of pyruvoyl-dependent arginine decarboxylase from Methanococcus jannaschii. AB - Pyruvoyl-dependent arginine decarboxylase (PvlArgDC) catalyzes the first step of the polyamine-biosynthetic pathway in plants and some archaebacteria. The pyruvoyl group of PvlArgDC is generated by an internal autoserinolysis reaction at an absolutely conserved serine residue in the proenzyme, resulting in two polypeptide chains. Based on the native structure of PvlArgDC from Methanococcus jannaschii, the conserved residues Asn47 and Glu109 were proposed to be involved in the decarboxylation and autoprocessing reactions. N47A and E109Q mutant proteins were prepared and the three-dimensional structure of each protein was determined at 2.0 A resolution. The N47A and E109Q mutant proteins showed reduced decarboxylation activity compared with the wild-type PvlArgDC. These residues may also be important for the autoprocessing reaction, which utilizes a mechanism similar to that of the decarboxylation reaction. PMID- 18391405 TI - Exploring structural variability in X-ray crystallographic models using protein local optimization by torsion-angle sampling. AB - Modeling structural variability is critical for understanding protein function and for modeling reliable targets for in silico docking experiments. Because of the time-intensive nature of manual X-ray crystallographic refinement, automated refinement methods that thoroughly explore conformational space are essential for the systematic construction of structurally variable models. Using five proteins spanning resolutions of 1.0-2.8 A, it is demonstrated how torsion-angle sampling of backbone and side-chain libraries with filtering against both the chemical energy, using a modern effective potential, and the electron density, coupled with minimization of a reciprocal-space X-ray target function, can generate multiple structurally variable models which fit the X-ray data well. Torsion angle sampling as implemented in the Protein Local Optimization Program (PLOP) has been used in this work. Models with the lowest R(free) values are obtained when electrostatic and implicit solvation terms are included in the effective potential. HIV-1 protease, calmodulin and SUMO-conjugating enzyme illustrate how variability in the ensemble of structures captures structural variability that is observed across multiple crystal structures and is linked to functional flexibility at hinge regions and binding interfaces. An ensemble-refinement procedure is proposed to differentiate between variability that is a consequence of physical conformational heterogeneity and that which reflects uncertainty in the atomic coordinates. PMID- 18391406 TI - Structures of two archaeal diphthine synthases: insights into the post translational modification of elongation factor 2. AB - The target of diphtheria toxin is the diphthamide residue in translation elongation factor 2 (EF-2), which is generated by a three-step post-translational modification of a specific histidine residue in the EF-2 precursor. In the second modification step, an S-adenosylmethionine-dependent methyltransferase, diphthine synthase (DS), catalyzes the trimethylation of the EF-2 precursor. The homodimeric crystal structures of the archaeal diphthine synthases from Pyrococcus horikoshii OT3 and Aeropyrum pernix K1 have been determined. These structures share essentially the same overall fold as the cobalt-precorrin-4 methyltransferase CbiF, confirming that DS belongs to the dimeric class III family of methyltransferases. In the P. horikoshii DS dimer, only one of the two active sites binds the reaction product S-adenosyl-L-homocysteine (AdoHcy), while the other active site contains no ligand. This asymmetric AdoHcy binding may be a consequence of intra-domain and inter-domain movements upon binding of AdoHcy at one of the two sites. These movements disrupt the twofold dimeric symmetry of the DS dimer and probably cause lower AdoHcy affinity at the other binding site. PMID- 18391407 TI - A mixture of fortunes: the curious determination of the structure of Escherichia coli BL21 Gab protein. AB - In protein crystallography, monodisperse protein samples of high purity are usually required in order to obtain diffraction-quality crystals. Here, crystals were reproducibly grown from a protein sample before its homogeneity had been determined. The sample was obtained after the first attempt to purify a recombinant target protein from an Escherichia coli cell lysate. Subsequent analysis revealed that it was a mixture of about 50 different proteins with no predominant species. Diffraction data were collected to 2.1 A and the space group was identified as I422. A molecular-replacement search with models of the expected target did not give a solution, which suggested that a contaminating E. coli protein had been crystallized. A PDB search revealed 256 structures determined in space group I422, of which 14 are E. coli proteins and two have unit-cell parameters similar to those observed. Molecular replacement with these structures showed a clear solution for one of them, the Gab protein. The structure is presented and compared with the deposited structure, from which it shows small but significant differences. The refined model contains bicine and sulfate as bound ligands, which provide insights into possible substrate-binding sites. PMID- 18391408 TI - A knowledge-driven approach for crystallographic protein model completion. AB - One of the most cumbersome and time-demanding tasks in completing a protein model is building short missing regions or ;loops'. A method is presented that uses structural and electron-density information to build the most likely conformations of such loops. Using the distribution of angles and dihedral angles in pentapeptides as the driving parameters, a set of possible conformations for the C(alpha) backbone of loops was generated. The most likely candidate is then selected in a hierarchical manner: new and stronger restraints are added while the loop is built. The weight of the electron-density correlation relative to geometrical considerations is gradually increased until the most likely loop is selected on map correlation alone. To conclude, the loop is refined against the electron density in real space. This is started by using structural information to trace a set of models for the C(alpha) backbone of the loop. Only in later steps of the algorithm is the electron-density correlation used as a criterion to select the loop(s). Thus, this method is more robust in low-density regions than an approach using density as a primary criterion. The algorithm is implemented in a loop-building program, Loopy, which can be used either alone or as part of an automatic building cycle. Loopy can build loops of up to 14 residues in length within a couple of minutes. The average root-mean-square deviation of the C(alpha) atoms in the loops built during validation was less than 0.4 A. When implemented in the context of automated model building in ARP/wARP, Loopy can increase the completeness of the built models. PMID- 18391410 TI - Structure of hepatitis C virus IRES subdomain IIa. AB - The hepatitis C (HCV) internal ribosome entry site (IRES) element plays a central role in cap-independent translation of the viral genomic RNA. The unique conformation of IRES domain II is critical for 80S ribosomal assembly and initiation of viral translation. Here, the crystal structure of subdomain IIa of the HCV IRES has been determined at 2.3 A resolution, revealing the positions of divalent metal ions and complex inter-strand interactions that stabilize the L shaped conformation of the RNA. The presence of divalent metal ions was necessary for crystal formation. Magnesium ions occupy specific sites that appear to be critical for the formation of the folded conformation. Subdomain IIa also was crystallized in the presence of strontium, which improved the diffraction quality of the crystals and the ability to identify interactions of the RNA with metal ions and tightly bound water molecules. The hinge region and noncanonical G-U base-pair motifs are stabilized by divalent metal ions and provide unique structural features that are potential interaction sites for small-molecule ligands. The information obtained from the crystal structure provides a basis for structure-guided design of HCV translation inhibitors targeting disruption of ribosomal assembly. PMID- 18391409 TI - A 7MUm mini-beam improves diffraction data from small or imperfect crystals of macromolecules. AB - A simple apparatus for achieving beam sizes in the range 5-10 MUm on a synchrotron beamline was implemented in combination with a small 125 x 25 MUm focus. The resulting beam had sufficient flux for crystallographic data collection from samples smaller than 10 x 10 x 10 MUm. Sample data were collected representing three different scenarios: (i) a complete 2.0 data set from a single strongly diffracting microcrystal, (ii) a complete and redundant 1.94 A data set obtained by merging data from six microcrystals and (iii) a complete 2.24 A data set from a needle-shaped crystal with less than 12 x 10 MUm cross-section and average diffracting power. The resulting data were of high quality, leading to well refined structures with good electron-density maps. The signal-to-noise ratios for data collected from small crystals with the mini-beam were significantly higher than for equivalent data collected from the same crystal with a 125 x 25 MUm beam. Relative to this large beam, use of the mini-beam also resulted in lower refined crystal mosaicities. The mini-beam proved to be advantageous for inhomogeneous large crystals, where better ordered regions could be selected by the smaller beam. PMID- 18391411 TI - Structure of peptidyl-tRNA hydrolase 2 from Pyrococcus horikoshii OT3: insight into the functional role of its dimeric state. AB - Peptidyl-tRNA hydrolases catalyze the hydrolytic removal of the peptidyl moiety from the peptidyl-tRNA molecule to prevent misreading during translation. Here, the expression, purification, crystallization and X-ray diffraction study of peptidyl-tRNA hydrolase 2 from Pyrococcus horikoshii OT3 (PhPth2) are described. The crystal structures were determined as similar biological dimers in two different forms: P4(1)2(1)2 at 1.2 A resolution (form 1) and P4(3)22 at 1.9 A resolution (form 2). In the form 1 structure, the asymmetric unit contains one PhPth2 subunit and a crystallographic twofold axis defines the dimeric association with the cognate subunit. In the form 2 structure, there are two PhPth2 subunits in the asymmetric unit that make a similar dimer with a noncrystallographic twofold axis. In order to evaluate the thermodynamic stability, the intra-protomer and inter-protomer interactions of PhPth2 were analyzed and compared with those of other Pth2-family members. The thermodynamic parameters show that the large number of ion pairs compared with family members from other mesophilic organisms would contribute to the thermostability of PhPth2. The structural difference between the two dimers was quantitatively evaluated by a multiple C(alpha)-atom superposition. A significant structural difference between the two dimers was observed around the putative active site of this enzyme. A rigid-body rotation takes place so as to retain the dimeric twofold symmetry, suggesting positive cooperativity upon tRNA binding. The mechanism of ligand binding was further investigated using a docking model with a tRNA molecule. The docking study suggests that the binding of tRNA requires its simultaneous interaction with both subunits of the PhPth2 dimer. PMID- 18391412 TI - Multiple crystal structures of actin dimers and their implications for interactions in the actin filament. AB - The structure of actin in its monomeric form is known at high resolution, while the structure of filamentous F-actin is only understood at considerably lower resolution. Knowing precisely how the monomers of actin fit together would lead to a deeper understanding of the dynamic behavior of the actin filament. Here, a series of crystal structures of actin dimers are reported which were prepared by cross-linking in either the longitudinal or the lateral direction in the filament state. Laterally cross-linked dimers, comprised of monomers belonging to different protofilaments, are found to adopt configurations in crystals that are not related to the native structure of filamentous actin. In contrast, multiple structures of longitudinal dimers consistently reveal the same interface between monomers within a single protofilament. The reappearance of the same longitudinal interface in multiple crystal structures adds weight to arguments that the interface visualized is similar to that in actin filaments. Highly conserved atomic interactions involving residues 199-205 and 287-291 are highlighted. PMID- 18391413 TI - Structure of acostatin, a dimeric disintegrin from Southern copperhead (Agkistrodon contortrix contortrix), at 1.7 A resolution. AB - Disintegrins are a family of small (4-14 kDa) proteins that bind to another class of proteins, integrins. Therefore, as integrin inhibitors, they can be exploited as anticancer and antiplatelet agents. Acostatin, an alphabeta heterodimeric disintegrin, has been isolated from the venom of Southern copperhead (Agkistrodon contortrix contortrix). The three-dimensional structure of acostatin has been determined by macromolecular crystallography using the molecular-replacement method. The asymmetric unit of the acostatin crystals consists of two heterodimers. The structure has been refined to an R(work) and R(free) of 18.6% and 21.5%, respectively, using all data in the 20-1.7 A resolution range. The structure of all subunits is similar and is well ordered into N-terminal and C terminal clusters with four intramolecular disulfide bonds. The overall fold consists of short beta-sheets, each of which is formed by a pair of antiparallel beta-strands connected by beta-turns and flexible loops of different lengths. Conformational flexibility is found in the RGD loops and in the C-terminal segment. The interaction of two N-terminal clusters via two intermolecular disulfide bridges anchors the alphabeta chains of the acostatin dimers. The C terminal clusters of the heterodimer project in opposite directions and form a larger angle between them in comparison with other dimeric disintegrins. Extensive interactions are observed between two heterodimers, revealing an alphabetabetaalpha acostatin tetramer. Further experiments are required to identify whether the alphabetabetaalpha acostatin complex plays a functional role in vivo. PMID- 18391414 TI - Deposition of diffraction images to be discussed at the Open Meeting of the Commission on Biological Macromolecules of the IUCr in Osaka. PMID- 18391415 TI - Structural analysis of the complex of Keap1 with a prothymosin alpha peptide. AB - The Nrf2 transcription factor, which plays important roles in oxidative and xenobiotic stress, is negatively regulated by the cytoplasmic repressor Keap1. The beta-propeller/Kelch domain of Keap1, which is formed by the double-glycine repeat and C-terminal region domains (Keap1-DC), interacts directly with the Neh2 domain of Nrf2. The nuclear oncoprotein prothymosin alpha (ProTalpha) also interacts directly with Keap1 and may play a role in the dissociation of the Keap1-Nrf2 complex. The structure of Keap1-DC complexed with a ProTalpha peptide (amino acids 39-54) has been determined at 1.9 A resolution. The Keap1-bound ProTalpha peptide possesses a hairpin conformation and binds to the Keap1 protein at the bottom region of the beta-propeller domain. Complex formation occurs as a consequence of their complementary electrostatic interactions. A comparison of the present structure with recently reported Keap1-DC complex structures revealed that the DLG and ETGE motifs of the Neh2 domain of Nrf2 and the ProTalpha peptide bind to Keap1 in a similar manner but with different binding potencies. PMID- 18391416 TI - Monoclinic form of isopentenyl diphosphate isomerase: a case of polymorphism in biomolecular crystals. AB - Type 1 isopentenyl diphosphate isomerase (IDI-1) has been crystallized in a new crystal form. After data collection from small thin needle-shaped crystals, a new monoclinic form of the studied protein was identified. In this article, the three crystal forms of IDI-1 (orthorhombic, monoclinic and trigonal) are compared. PMID- 18391417 TI - Structure of the SH3 domain of rat endophilin A2. AB - The crystal structure of the SH3 domain of rat endophilin A2 has been determined by the multiwavelength anomalous dispersion method and refined at a resolution of 1.70 A to R and R(free) values of 0.196 and 0.217, respectively. The structure adheres to the canonical SH3-domain fold and is highly similar to those of the corresponding domains of endophilins A1 and A3. An intermolecular packing interaction between two molecules in the lattice exploits features that are commonly observed in SH3-domain ligand recognition, including the insertion of a proline side chain into the ligand-binding groove of the protein and the recognition of a basic residue by a cluster of acidic side chains on the RT loop. PMID- 18391418 TI - Structure of the cold-shock domain protein from Neisseria meningitidis reveals a strand-exchanged dimer. AB - The structure of the cold-shock domain protein from Neisseria meningitidis has been solved to 2.6 A resolution and shown to comprise a dimer formed by the exchange of two beta-strands between protein monomers. The overall fold of the monomer closely resembles those of other bacterial cold-shock proteins. The neisserial protein behaved as a monomer in solution and was shown to bind to a hexathymidine oligonucleotide with a stoichiometry of 1:1 and a K(d) of 1.25 microM. PMID- 18391419 TI - Crystallization and preliminary X-ray crystallographic analysis of a carbonyl reductase from Candida parapsilosis. AB - A novel short-chain NADPH-dependent (S)-1-phenyl-1,2-ethanediol dehydrogenase (SCR) has been crystallized. Two distinct but related crystal forms of SCR were obtained using the hanging-drop vapour-diffusion method and a reservoir solution consisting of 18%(w/v) polyethylene glycol 2000 monomethyl ether and 8%(v/v) 2 propanol as the precipitant. The crystals were rhomboid in shape with average dimensions of 0.3 x 0.3 x 0.4 mm and diffracted to a resolution of 2.7-3.0 A. The crystal forms both belong to space group P2(1)2(1)2(1) and have unit-cell parameters a = 104.7, b = 142.8, c = 151.8 A and a = 101.1, b = 146.0, c = 159.8 A. The calculated values of V(M), rotation-function and translation-function solutions and consideration of potential crystal packing suggest that there are eight protein subunits per asymmetric unit. PMID- 18391420 TI - Crystallization and preliminary X-ray diffraction analysis of the glucuronoyl esterase catalytic domain from Hypocrea jecorina. AB - The catalytic domain of the glucuronoyl esterase from Hypocrea jecorina (anamorph Trichoderma reesei) was overexpresssed, purified and crystallized by the sitting drop vapor-diffusion method using 1.4 M sodium/potassium phosphate pH 6.9. The crystals belonged to space group P2(1)2(1)2(1) and X-ray diffraction data were collected to 1.9 A resolution. This is the first enzyme with glucoronoyl esterase activity to be crystallized; its structure will be valuable in lignocellulose degradation research. PMID- 18391421 TI - Expression, purification and crystallization of a lyssavirus matrix (M) protein. AB - The matrix (M) proteins of lyssaviruses (family Rhabdoviridae) are crucial to viral morphogenesis as well as in modulating replication and transcription of the viral genome. To date, no high-resolution structural information has been obtained for full-length rhabdovirus M. Here, the cloning, expression and purification of the matrix proteins from three lyssaviruses, Lagos bat virus (LAG), Mokola virus and Thailand dog virus, are described. Crystals have been obtained for the full-length M protein from Lagos bat virus (LAG M). Successful crystallization depended on a number of factors, in particular the addition of an N-terminal SUMO fusion tag to increase protein solubility. Diffraction data have been recorded from crystals of native and selenomethionine-labelled LAG M to 2.75 and 3.0 A resolution, respectively. Preliminary analysis indicates that these crystals belong to space group P6(1)22 or P6(5)22, with unit-cell parameters a = b = 56.9-57.2, c = 187.9-188.6 A, consistent with the presence of one molecule per asymmetric unit, and structure determination is currently in progress. PMID- 18391422 TI - Structural analysis of bacteriophage-encoded peptidoglycan hydrolase domain KMV36C: crystallization and preliminary X-ray diffraction. AB - The C-terminus of gp36 of bacteriophage varphiKMV (KMV36C) functions as a particle-associated muramidase, presumably as part of the injection needle of the phiKMV genome during infection. Crystals of KMV36C were obtained by hanging-drop vapour diffusion and diffracted to a resolution of 1.6 A. The crystals belong to the cubic space group P432, with unit-cell parameters a = b = c = 102.52 A. KMV36C shows 30% sequence identity to T4 lysozyme (PDB code 1l56). PMID- 18391423 TI - Crystallization and preliminary X-ray diffraction analysis of human IL-22 bound to the extracellular IL-22R1 chain. AB - Interleukin-22 (IL-22) is a potent mediator of cellular inflammatory responses. Crystals of IL-22 bound to the extracellular high-affinity cell-surface receptor sIL-22R1 have been grown from polyethylene glycol solutions. Crystals suitable for X-ray diffraction analysis were only obtained with mutants of IL-22 and sIL 22R1 that removed the N-linked glycosylation sites found in the wild-type amino acid sequences. The crystals belonged to space group P2(1), with unit-cell parameters a = 50.43, b = 76.33, c = 114.92 A, beta = 92.45 degrees , and diffracted X-rays to 3.2 A resolution. The crystallographic asymmetric unit contained two IL-22-sIL-22R1 complexes, corresponding to a solvent content of approximately 52%. PMID- 18391424 TI - Preliminary time-of-flight neutron diffraction study of human deoxyhemoglobin. AB - Human hemoglobin (HbA) is an intricate system that has evolved to efficiently transport oxygen molecules (O(2)) from lung to tissue. Its quaternary structure can fluctuate between two conformations, T (tense or deoxy) and R (relaxed or oxy), which have low and high affinity for O(2), respectively. The binding of O(2) to the heme sites of HbA is regulated by protons and by inorganic anions. In order to investigate the role of the protonation states of protein residues in O(2) binding, large crystals of deoxy HbA (approximately 20 mm(3)) were grown in D(2)O under anaerobic conditions for neutron diffraction studies. A time-of flight neutron data set was collected to 1.8 A resolution on the Protein Crystallography Station (PCS) at the spallation source run by Los Alamos Neutron Science Center (LANSCE). The HbA tetramer (64.6 kDa; 574 residues excluding the four heme groups) occupies the largest asymmetric unit (space group P2(1)) from which a high-resolution neutron data set has been collected to date. PMID- 18391425 TI - Crystallization and preliminary crystallographic analysis of the second GAF domain of DevS from Mycobacterium smegmatis. AB - Mycobacterium tuberculosis is known to transform into the nonreplicating persistence state under the influence of hypoxia or nitric oxide. DevS-DevR is a two-component regulatory system that mediates the genetic response for the transformation. DevS is a histidine kinase that contains two GAF domains for sensing hypoxia or nitric oxide. The second GAF from M. smegmatis DevS was crystallized using the sitting-drop vapour-diffusion method in the presence of sodium citrate and 2-propanol as precipitants. X-ray diffraction data were collected from crystals containing selenomethionine to a maximum resolution of 2.0 A on a synchrotron beamline. The crystals belong to the hexagonal space group P6(1). The asymmetric unit contains one molecule, corresponding to a packing density of 2.5 A(3) Da(-1). The selenium substructure was determined by the single anomalous dispersion method and structure refinement is in progress. PMID- 18391426 TI - Cloning, purification and preliminary X-ray analysis of the C-terminal domain of Helicobacter pylori MotB. AB - The C-terminal domain of MotB (MotB-C) contains a putative peptidoglycan-binding motif and is believed to anchor the MotA/MotB stator unit of the bacterial flagellar motor to the cell wall. Crystals of Helicobacter pylori MotB-C (138 amino-acid residues) were obtained by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. These crystals belong to space group P2(1), with unit-cell parameters a = 50.8, b = 89.5, c = 66.3 A, beta = 112.5 degrees . The crystals diffract X-rays to at least 1.6 A resolution using a synchrotron-radiation source. Self-rotation function and Matthews coefficient calculations suggest that the asymmetric unit contains one tetramer with 222 point-group symmetry. The anomalous difference Patterson maps calculated for an ytterbium-derivative crystal using diffraction data at a wavelength of 1.38 A showed significant peaks on the v = 1/2 Harker section, suggesting that ab initio phase information could be derived from the MAD data. PMID- 18391427 TI - Cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from Thalictrum flavum. AB - Norcoclaurine synthase (NCS) catalyzes the condensation of 3,4 dihydroxyphenylethylamine (dopamine) and 4-hydroxyphenylacetaldehyde (4-HPAA) as the first committed step in the biosynthesis of benzylisoquinoline alkaloids in plants. The protein was cloned, expressed and purified. Crystals were obtained at 294 K by the hanging-drop vapour-diffusion method using ammonium sulfate and sodium chloride as precipitant agents and diffract to better than 3.0 A resolution using a synchrotron-radiation source. The crystals belong to the trigonal space group P3(1)21, with unit-cell parameters a = b = 86.31, c = 118.36 A. A selenomethionine derivative was overexpressed, purified and crystallized in the same space group. A complete MAD data set was collected at 2.7 A resolution. The model is under construction. PMID- 18391428 TI - Crystallization and preliminary crystallographic analysis of the transpeptidase domain of penicillin-binding protein 2B from Streptococcus pneumoniae. AB - Penicillin-binding protein (PBP) 2B from Streptococcus pneumoniae catalyzes the cross-linking of peptidoglycan precursors that occurs during bacterial cell-wall biosynthesis. A selenomethionyl (SeMet) substituted PBP 2B transpeptidase domain was isolated from a limited proteolysis digest of a soluble form of recombinant PBP 2B and then crystallized. The crystals belonged to space group P4(3)2(1)2, with unit-cell parameters a = b = 86.39, c = 143.27 A. Diffraction data were collected to 2.4 A resolution using the BL32B2 beamline at SPring-8. The asymmetric unit contains one protein molecule and 63.7% solvent. PMID- 18391429 TI - Crystallization and preliminary X-ray diffraction analysis of the multidrug efflux transporter NorM from Neisseria gonorrhoeae. AB - The crystallization and preliminary X-ray data analysis of the NorM multidrug efflux pump produced by Neisseria gonorrhoeae are reported. NorM is a cytoplasmic membrane protein that consists of 459 amino-acid residues. It is a member of the recently classified multidrug and toxic compound extrusion (MATE) family of transporters and recognizes a number of cationic toxic compounds such as ethidium bromide, acriflavin, 2-N-methylellipticinium and ciprofloxacin. Recombinant NorM protein was expressed in Escherichia coli and purified by metal-affinity and gel filtration chromatography. The protein was crystallized using hanging-drop vapor diffusion. X-ray diffraction data were collected from cryocooled crystals at a synchrotron light source. The best crystal diffracted anisotropically to 3.8 A and diffraction data were complete to 6.5 A resolution. The space group was determined to be C2, with unit-cell parameters a = 81.5, b = 164.4, c = 111.5 A. PMID- 18391430 TI - Crystallization and preliminary X-ray diffraction studies of Seneca Valley virus 001, a new member of the Picornaviridae family. AB - Seneca Valley Virus-001 (SVV-001) is a newly found species in the Picornaviridae family. SVV-001 is the first naturally occurring nonpathogenic picornavirus observed to mediate selective cytotoxicity towards tumor cells with neuroendocrine cancer features. The nonsegmented (+)ssRNA genome of SVV-001 shares closest sequence similarity to the genomes of the members of the Cardiovirus genus. However, based on the distinct characteristics of the genome organization and other biochemical properties, it has been suggested that SVV-001 represents a new genus, namely 'Senecavirus', in the Picornaviridae family. In order to understand the oncolytic properties of SVV-001, the native virus was crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space group R3, with unit-cell parameters (in the hexagonal setting) a = b = 311.5, c = 1526.4 A. Although the SVV crystals diffracted to better than 2.3 A resolution, the data quality is acceptable [I/sigma(I) > 2.0] to 2.6 A resolution. The unit-cell volume and the locked rotation-function analysis suggest that six particles could be accommodated in the unit cell, with two distinct sets of one third of a particle, each containing 20 protomers, occupying the crystallographic asymmetric unit. (ClinicalTrials.gov identifier NCT00314925) PMID- 18391431 TI - Crystallization of an Lhx3-Isl1 complex. AB - A stable intramolecular complex comprising the LIM domains of the LIM-homeodomain protein Lhx3 tethered to a peptide region of Isl1 has been engineered, purified and crystallized. The monoclinic crystals belong to space group C2, with unit cell parameters a = 119, b = 62.2, c = 51.9 A, beta = 91.6 degrees , and diffract to 2.05 A resolution. PMID- 18391432 TI - Preliminary structural studies on the MtxX protein from Methanococcus jannaschii. AB - Methanococcus jannaschii has an mtr gene cluster expressing N(5) methyltetrahydromethanopterin:coenzyme M methyltransferase, which generates methane by reducing CO(2) with H(2) with concomitant energy production under strictly anaerobic conditions. Some methanogenic archaea also have an mtr gene cluster homologue, the mtxXAH gene cluster. M. jannaschii has both an entire mtr gene cluster and a single mtxX gene instead of the whole mtxXAH gene cluster. A PSI-BLAST search, secondary-structure prediction and the absence of phosphotransacetylase activity in M. jannaschii strongly support the possibility that the MtxX protein constitutes a unique methyltransferase family. In this study, the MtxX protein from M. jannaschii has been cloned, expressed, purified and crystallized. Synchrotron data were collected to 2.9 A from a crystal of selenomethionine-substituted MtxX protein. The crystal belonged to the primitive hexagonal space group P6(1)22, with unit-cell parameters a = 54.9, b = 54.9, c = 341.1 A, beta = 120.0 degrees . A full structure determination is under way in order to provide insight into the structure-function relationship of this protein. PMID- 18391433 TI - Crystallization and preliminary crystallographic analysis of a plant type III polyketide synthase that produces benzalacetone. AB - Benzalacetone synthase (BAS) from Rheum palmatum is a plant-specific type III polyketide synthase that catalyzes the one-step decarboxylative condensation of 4 coumaroyl-CoA with malonyl-CoA to produce the diketide 4-(4-hydroxyphenyl)-but-3 en-2-one. Recombinant BAS expressed in Escherichia coli was crystallized by the sitting-drop vapour-diffusion method. The crystals belong to space group P2(1), with unit-cell parameters a = 54.6, b = 89.6, c = 81.1 A, alpha = gamma = 90.0, beta = 100.5 degrees . Diffraction data were collected to 1.8 A resolution using synchrotron radiation at BL24XU of SPring-8. PMID- 18391434 TI - Crystallization and preliminary X-ray analysis of the complex of the first von Willebrand type C domain bound to bone morphogenetic protein 2. AB - Crossveinless 2 (CV2) is a member of the chordin family, a protein superfamily that modulates the activity of bone morphogenetic proteins such as BMP2. The BMPs represent a large group of secreted proteins that control many steps during embryonal development and in tissue and organ homeostasis in the adult organism. The gene encoding the first von Willebrand type C domain (VWC1) of CV2 was cloned, expressed in Escherichia coli and purified to homogeneity. The binary complex of CV2 VWC1 and BMP2 was purified and subjected to crystallization. Crystals of SeMet-labelled proteins were obtained in two different forms belonging to the tetragonal space groups P4(1)2(1)2 and I4(1), with unit-cell parameters a = b = 86.7, c = 139.2 A and a = b = 83.7, c = 139.6 A, respectively. Initial analysis suggests that a complete binary complex consisting of one BMP2 dimer bound to two CV2 VWC1 domains is present in the asymmetric unit. PMID- 18391435 TI - Crystallization of hepatocyte nuclear factor 4 alpha (HNF4 alpha) in complex with the HNF1 alpha promoter element. AB - Hepatocyte nuclear factor 4alpha (HNF4alpha) is a member of the nuclear receptor superfamily that plays a central role in organ development and metabolic functions. Mutations on HNF4alpha cause maturity-onset diabetes of the young (MODY), a dominant monogenic cause of diabetes. In order to understand the molecular mechanism of promoter recognition and the molecular basis of disease causing mutations, the recombinant HNF4alpha DNA-binding domain was prepared and used in a study of its binding properties and in crystallization with a 21-mer DNA fragment that contains the promoter element of another MODY gene, HNF1alpha. The HNF4alpha protein displays a cooperative and specific DNA-binding activity towards its target gene-recognition elements. Crystals of the complex diffract to 2.0 A using a synchrotron-radiation source under cryogenic (100 K) conditions and belong to space group C2, with unit-cell parameters a = 121.63, b = 35.43, c = 70.99 A, beta = 119.36 degrees . A molecular-replacement solution has been obtained and structure refinement is in progress. This structure and the binding studies will provide the groundwork for detailed functional and biochemical studies of the MODY mutants. PMID- 18391436 TI - Crystallization and preliminary X-ray diffraction analysis of recombinant hepatitis E virus-like particle. AB - Hepatitis E virus (HEV) accounts for the majority of enterically transmitted hepatitis infections worldwide. Currently, there is no specific treatment for or vaccine against HEV. The major structural protein is derived from open reading frame (ORF) 2 of the viral genome. A potential oral vaccine is provided by the virus-like particles formed by a protein construct of partial ORF3 protein (residue 70-123) fused to the N-terminus of the ORF2 protein (residues 112-608). Single crystals obtained by the hanging-drop vapour-diffusion method at 293 K diffract X-rays to 8.3 A resolution. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 337, b = 343, c = 346 A, alpha = beta = gamma = 90 degrees , and contain one particle per asymmetric unit. PMID- 18391439 TI - Effect of a novel ascorbic derivative, disodium isostearyl 2-O-L-ascorbyl phosphate, on normal human dermal fibroblasts against reactive oxygen species. AB - The novel amphiphilic vitamin C derivative disodium isostearyl 2-O-L-ascorbyl phosphate (VCP-IS-2Na), which has a C(18) alkyl chain attached to the stable ascorbate derivative sodium L-ascorbic acid 2-phosphate (VCP-Na), was evaluated for reduction of cell damage induced by oxidative stress, ultraviolet A (UVA), ultraviolet B (UVB), and H(2)O(2); stimulation of collagen synthesis against UVA irradiation; and inhibition of matrix metalloproteinase-1 (MMP-1) activity induced by UVA in human normal dermal fibroblasts. VCP-IS-2Na pretreatment resulted in significant protection against cell damage induced by UVB, UVA, and H(2)O(2). The amount of type I collagen following UVA irradiation was increased by treatment with VCP-IS-2Na in a concentration-dependent manner. These effects of VCP-IS-2Na were superior to those of L-ascorbic acid (vitamin C, VC) and VCP Na. On the other hand, VCP-IS-2Na suppressed 65% of the excess MMP-1 irradiated UVA, and VC and VCP-Na slightly suppressed it. PMID- 18391437 TI - Expression, purification, crystallization and preliminary X-ray diffraction analysis of thioredoxin Trx1 from Saccharomyces cerevisiae. AB - Thioredoxins play key roles in the cellular response to oxidative stress. Three isoforms of thioredoxin have been identified in Saccharomyces cerevisiae: two that are cytosolic (Trx1 and Trx2) and one that is mitochondrial (Trx3). In the present work, the cytosolic form Trx1 was cloned, expressed, purified and crystallized. Crystals were obtained by the hanging-drop vapour-diffusion method. A data set was collected from a single crystal to 1.7 A resolution. The crystal belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 32.29, b = 46.59, c = 64.20 A, alpha = beta = gamma = 90 degrees . PMID- 18391440 TI - NblR is a novel one-component response regulator in the cyanobacterium Synechococcus elongatus PCC 7942. AB - A response regulator, NblR, of the cyanobacterium Synechococcus elongatus PCC 7942 is known to induce expression of the nblA gene, a key factor in phycobilisome degradation (bleaching) under nutrient-deprivation conditions. In this study, we observed phosphorylation-independent regulation of NblR activity. We constructed a mutant strain expressing NblR (D57A), in which a putative phospho-accepting Asp-57 was replaced with Ala residue. Under nitrogen deprivation, this strain exhibited the typical bleaching phenotype observed in wild-type cells. Moreover, in the mutant, the nblA transcript accumulated at a level similar to that of the wild type. Our results indicate that activation of NblR is independent of phosphorylation, if any, by a cognate histidine kinase. Screening of proteins interacting with NblR by yeast two-hybrid analysis revealed two candidates, MreC and NarB, suggesting a novel mechanism that activates NblR, or other functions of the response regulator. PMID- 18391441 TI - Anti-hyperglycemic activity of kiwifruit leaf (Actinidia deliciosa) in mice. AB - The methanol extract of kiwifruit leaf suppressed the postprandial blood glucose level after an oral administration of soluble starch or sucrose in mice. The mechanism of action is proposed to be due to the alpha-amylase-inhibiting activity in the 90% aqueous methanol fraction and alpha-glucosidase-inhibiting activity in the n-buthanol fraction, based on the results of in vitro experiments. PMID- 18391442 TI - A new family of D-2-hydroxyacid dehydrogenases that comprises D-mandelate dehydrogenases and 2-ketopantoate reductases. AB - The gene for the D-mandelate dehydrogenase (D-ManDH) of Enterococcus faecalis IAM10071 was isolated by means of an activity staining procedure and PCR and expressed in Escherichia coli cells. The recombinant enzyme exhibited high catalytic activity toward various 2-ketoacid substrates with bulky hydrophobic side chains, particularly C3-branched substrates such as benzoylformate and 2 ketoisovalerate, and strict coenzyme specificity for NADH and NAD(+). It showed marked sequence similarity with known NADP-dependent 2-ketopantoate reductases (KPR). These results indicate that together with KPR, D-ManDH constitutes a new family of D-2-hydroxyacid dehydrogenases that act on C3-branched 2-ketoacid substrates with various specificities for coenzymes and substrates. PMID- 18391443 TI - Expression of parsley flavone synthase I establishes the flavone biosynthetic pathway in Arabidopsis thaliana. AB - Arabidopsis thaliana lacks the flavone biosynthetic pathway, probably because of a lack or low activity of a flavone synthase. To establish this biosynthetic pathway in Arabidopsis, we subjected this model plant to transformation with the parsley gene for flavone synthase type I (FNS-I). Transgenic seedlings expressing FNS-I were cultured in liquid medium with or without naringenin, and plant extracts were then analyzed by high-performance liquid chromatography. In contrast to wild-type seedlings, the transgenic seedlings accumulated substantial amounts of apigenin, which is produced from naringenin by FNS-I, and the apigenin level correlated with the abundance of FNS-I mRNA in three different transgenic lines. These results indicate that the FNS-I transgene produces a functional enzyme that catalyzes the conversion of naringenin to apigenin in Arabidopsis. These FNS-I transgenic lines should prove useful in investigating the in vivo functions of enzymes that mediate the synthesis of the wide variety of flavones found in other plants. PMID- 18391444 TI - Identification of Saccharomyces cerevisiae Tub1 alpha-tubulin as a potential target for NKH-7, a cytotoxic 1-naphthol derivative compound. AB - In a screening for small-molecule compounds that alleviate the deleterious effects of external CaCl(2) on zds1 Delta strain yeast, we found 2-((1 (hydroxymethyl) cyclohexyl) methyl) naphthalen-1-ol (NKH-7) to be an active compound. NKH-7 also inhibited cell growth at higher concentrations. To identify its target in growth inhibition, we isolated NKH-7-resistant mutants and selected those mutants that exhibited dominant or semi-dominant resistance specifically to NKH-7. By gene cloning, a TUB1 mutant gene encoding alpha-tubulin with a Ser248Pro mutation was identified. Deletion of the TUB3 gene, a minor gene encoding alpha-tubulin, led to supersensitivity to NKH-7. Cellular tubulin containing arrays as visualized by green fluorescent protein (GFP)-labeled alpha tubulin diminished rapidly on exposure to the inhibitor. The mutation was situated proximal to the alpha-beta interface of alpha-tubulin in microtubule protofilaments, suggesting the possibility that NKH-7 affects the hydrolysis of GTP bound to beta-tubulin. A functional connection perhaps exists between the tubulin inhibition and Ca(2+)-dependent cell-cycle regulation. PMID- 18391445 TI - Structure-antibacterial activity relationship for 9-O,9'-O-demethyl (+) virgatusin. AB - The relationship between the antibacterial activity and structure of 9-O,9'-O demethyl (+)-virgatusin (Virg 3) was examined. The conversion of hydroxy groups on the 9 and 9' positions to amino groups increased the activity. It was found that the 3'-methoxy group was more important for higher activity than the 4' methoxy group on the 7'-phenyl group, and that the 3,4-methylenedioxy group on the 7-phenyl group was necessary for activity. PMID- 18391446 TI - Involvement of LuxS in the regulation of motility and flagella biogenesis in Vibrio alginolyticus. AB - The fish pathogen Vibrio alginolyticus contains two unique flagellar systems. The LuxS quorum sensing system is reported to regulate the expression of virulence factors in a wide variety of pathogenic bacteria. Our previous work demonstrated that inactive luxS led to decreased virulence in V. alginolyticus. In this study, LuxS-dependent regulation of motility and flagella biogenesis, the potential virulence factors in V. alginolyticus, were further investigated. A luxS-deleted mutant showed deficiency in motility and flagella formation, and an intact luxS complemented the defect. Since motility is flagella dependent, V. alginolyticus flagella biogenesis related genes, including the flagella regulator genes flaK and lafK and the sub-hierarchical flagellar genes fliS and lafA, were cloned and identified. Based on quantitative real-time reverse transcription-PCR, differential expression of these genes was confirmed in wild-type and luxS mutants. Our results indicate that LuxS plays an important role in the regulation of motility and flagella biogenesis in V. alginolyticus. PMID- 18391447 TI - Characterization of two compatible small plasmids from Sphingobium yanoikuyae. AB - We isolated and characterized two small cryptic indigenous plasmids, pYAN-1 (4,896 bp) and pYAN-2 (4,687 bp), from Sphingobium yanoikuyae, and developed a versatile system that permitted genetic manipulation of the genus Sphingomonas. Nucleotide sequencing of both plasmids revealed that they contained mobA, mobs, and repA genes, which are predicted to encode proteins associated with mobilization and replication, in common. Transformation with each plasmid harboring the antibiotic resistance gene by electroporation was fully successful, using Novosphingobium capsulatum as a host. PMID- 18391448 TI - Energy metabolism of a unique acetic acid bacterium, Asaia bogorensis, that lacks ethanol oxidation activity. AB - Acetic acid bacteria (AAB) are known as a vinegar producer on account of their ability to accumulate a high concentration of acetic acid due to oxidative fermentation linking the ethanol oxidation respiratory chain. Reactions in oxidative fermentation cause poor growth because a large amount of the carbon source is oxidized incompletely and the harmful oxidized products are accumulated almost stoichiometrically in the culture medium during growth, but a newly identified AAB, Asaia, has shown unusual properties, including scanty acetic acid production and rapid growth, as compared with known AAB as Acetobacter, Gluconobacter, and Gluconacetobacter. To understand these unique properties of Asaia in more detail, the respiratory chain and energetics of this strain were investigated. It was found that Asaia lacks quinoprotein alcohol dehydrogenase, but has other sugar and sugar alcohol-oxidizing enzymes specific to the respiratory chain of Gluconobacter, especially quinoprotein glycerol dehydrogenase. It was also found that Asaia has a cyanide-sensitive cytochrome bo(3)-type ubiquinol oxidase as sole terminal oxidase in the respiratory chain, and that it exhibits a higher H(+)/O ratio. PMID- 18391449 TI - A rapid, simple, and effective method of constructing a randomly mutagenized plasmid library free from ligation. AB - The QuikChange site-directed mutagenesis methodology was applied to constructing a randomly mutagenized plasmid library simply by adding manganese to the reaction mixture. This method is superior to the normally employed Pol I-type polymerase based error-prone PCR in that (i) it does not require a subsequent ligation reaction, and (ii) there is no accumulation of mutations at the same site. alpha Complementation analysis and subsequent sequence analyses of the lacZ alpha genes in the mutated library revealed that the mutations occurred randomly within the target gene and involved all possible base substitutions. PMID- 18391450 TI - Characterization of a dihydrolipoyl dehydrogenase having diaphorase activity of Clostridium kluyveri. AB - The Clostridium kluyveri bfmBC gene encoding a putative dihydrolipoyl dehydrogenase (DLD; EC 1.8.1.4) was expressed in Escherichia coli, and the recombinant enzyme rBfmBC was characterized. UV-visible absorption spectrum and thin layer chromatography analysis of rBfmBC indicated that the enzyme contained a noncovalently but tightly attached FAD molecule. rBfmBC catalyzed the oxidation of dihydrolipoamide (DLA) with NAD(+) as a specific electron acceptor, and the apparent K(m) values for DLA and NAD(+) were 0.3 and 0.5 mM respectively. In the reverse reaction, the apparent K(m) values for lipoamide and NADH were 0.42 and 0.038 mM respectively. Like other DLDs, this enzyme showed NADH dehydrogenase (diaphorase) activity with some synthetic dyes, such as 2,6 dichlorophenolindophenol and nitro blue tetrazolium. rBfmBC was optimally active at 40 degrees C at pH 7.0, and the enzyme maintained some activity after a 30-min incubation at 60 degrees C. PMID- 18391451 TI - Structure and function of NAD kinase and NADP phosphatase: key enzymes that regulate the intracellular balance of NAD(H) and NADP(H). AB - The functions of NAD(H) (NAD(+) and NADH) and NADP(H) (NADP(+) and NADPH) are undoubtedly significant and distinct. Hence, regulation of the intracellular balance of NAD(H) and NADP(H) is important. The key enzymes involved in the regulation are NAD kinase and NADP phosphatase. In 2000, we first succeeded in identifying the gene for NAD kinase, thereby facilitating worldwide studies of this enzyme from various organisms, including eubacteria, archaea, yeast, plants, and humans. Molecular biological study has revealed the physiological function of this enzyme, that is to say, the significance of NADP(H), in some model organisms. Structural research has elucidated the tertiary structure of the enzyme, the details of substrate-binding sites, and the catalytic mechanism. Research on NAD kinase also led to the discovery of archaeal NADP phosphatase. In this review, we summarize the physiological functions, applications, and structure of NAD kinase, and the way we discovered archaeal NADP phosphatase. PMID- 18391452 TI - Proposed oxidative metabolic pathway for polypropylene glycol in Sphingobium sp. strain PW-1. AB - Polypropylene glycol (PPG)-assimilating Sphingobium sp. strain PW-1 was grown on 0.5% PPG 700. PPG and its metabolites were analyzed by MALDI-TOF mass spectrometry. An oxidized form of a terminal alcohol group appeared with each molecular species as a metabolite. Cell-free extract showed PPG dehydrogenase activity. In this way, the oxidative metabolic pathway for PPG was confirmed. PMID- 18391453 TI - Inhibitory effects of whisky congeners on melanogenesis in mouse B16 melanoma cells. AB - We examined the effect of whisky congeners, substances other than ethanol in whisky, on melanogenesis in mouse B16 melanoma cells. Treatment with whisky congeners significantly blocked melanogenesis. Our results indicate that the inhibitory effects of whisky congeners on melanogenesis is due to direct inhibition of tyrosinase activity and to suppression of tyrosinase protein levels. PMID- 18391454 TI - Biocalorimetric studies of the metabolic activity of Pseudomonas aeruginosa aerobically grown in a glucose-limited complex growth medium. AB - Biocalorimetric experiments were performed to investigate the aerobic growth of Pseudomonas aeruginosa, isolated from tannery saline wastewater. Growth factors (pH, Inoculum size, carbon source, temperature, aeration rate, and agitation rate) were optimized in shaker and calorimeter based on the growth of P. aeruginosa and heat generation rates. A limiting value of 0.2% glucose concentration was found to be optimum for the growth of P. aeruginosa in a complex growth medium, and the heat flux (q(r)) profiles resulting from the metabolic activity of P. aeruginosa further confirmed this observation. The bacterial growth profile was found to correlate well with the metabolic heat generated. Heat-yield values were calculated for both glucose consumption and the growth of P. aeruginosa from the calorimetric results. Metabolic shifts in substrate uptake from glucose to peptone present in growth medium was observed by the variations in heat-flux profile. The calorimetric data presented in this study should be useful in understanding the behavior of the isolated bacterial strain in degrading complex and mixed substrates commonly observed in tannery saline waste stream, and further to extend the results for scale-up studies. PMID- 18391455 TI - Synthesis of a series of fructooligosaccharides with sucrose and cycloinulohexaose extending over ten degrees of polymerization using cycloinulooligosaccharide fructanotransferase from Bacillus circulans OKUMZ 31B. AB - Prolonged incubation with sucrose as an acceptor and cycloinulohexaose as a donor at a high concentration of cycloinulooligosaccharide fructanotransferase afforded a series of fructooligosaccharides. Their chain-length distribution extended over 10 degrees of polymerization when the donor concentration was increased to 120 mM. Increasing the acceptor concentration proved effective in improving the yield of inulin-type oligosaccharides because hydrolysis was suppressed. PMID- 18391456 TI - Possible involvement of prolactin in the synthesis of lactoferrin in bovine mammary epithelial cells. AB - Bovine mammary epithelial cells (bMECs) synthesize lactoferrin, which is secreted into milk. Our results suggest that prolactin stimulated secretion of lactoferrin in primary bMECs and their clonal cell line under serum-free conditions. Prolactin also stimulated mRNA expression of lactoferrin in the clonal cell line. This effect was reduced by AG-490, suggesting that the prolactin-stimulated mRNA expression of lactoferrin was mediated by Janus kinase (JAK)2. PMID- 18391457 TI - Nuclear swelling occurs during premature senescence mediated by MAP kinases in normal human fibroblasts. AB - Excess thymidine induced premature senescence in normal human fibroblasts (TIG 7), with induction of typical senescence markers. Nuclear swelling, as well as cell swelling, was clearly observed in these senescent cells. Simultaneous addition of MAP kinase inhibitors, U0126, SB203580, and SP60025, effectively suppressed induction of premature senescence and senescence markers. PMID- 18391458 TI - A ribosome assembly factor Ebp2p, the yeast homolog of EBNA1-binding protein 2, is involved in the secretory response. AB - We have found that Ebp2p is essential for maturation of 25S rRNA and assembly of 60S pre-ribosomal subunits in Saccharomyces cerevisiae. We obtained three temperature-sensitive ebp2 mutants by PCR. Polysome analysis revealed that the synthesis of 60S ribosomal subunits was compromised in each of the ebp2 mutants at the restrictive temperature. The ebp2 alleles affected the transcriptional repression of both rRNA and ribosomal protein genes due to a secretion block. Fluorescence microscopy showed that a secretion block led to condensation of nucleolar Ebp2p, whereas that was not the case with the ebp2 mutant. These results suggest that Ebp2p is implicated in the secretory response, including changes in nucleolar architecture. PMID- 18391459 TI - Identification of new amine acceptor protein substrate candidates of transglutaminase in rat liver extract: use of 5-(biotinamido) pentylamine as a probe. AB - Transglutaminases (TGs) are a family of enzymes that catalyze Ca(2+)-dependent post-translational modification of proteins by introducing protein-protein crosslinks (between specific glutamine and lysine residues), amine incorporation, and site-specific deamidation. In this study, new amine acceptor protein substrates of TG were isolated from rat liver extract and identified using 5 (biotinamido) pentylamine, a biotinylated primary amine substrate, as a probe. TG protein substrate candidates labeled with biotin by endogenous TG activity were isolated and recovered by avidin column chromatography. Proteins with molecular masses of 40, 42, and 45 kDa were the main components of the labeled proteins. Determination of their partial amino acid sequences and immunoblotting analyses were done to identify them. The 45-kDa protein was identical with betaine homocysteine S-methyltransferase (EC 2.2.2.5), which was identified in our previous study. The 40- and 42-kDa proteins were identified as arginase-I (EC 3.5.3.1) and fructose-1,6-bisphosphatase (EC 3.1.3.11) respectively. TG catalyzed incorporation of 5-(biotinamido) pentylamine into both arginase-I and fructose 1,6-bisphosphatase purified from rat liver was confirmed in vitro. These results suggest that these two enzymes are the new protein substrate candidates of TG and that they can be modified post-translationally by cellular TG. PMID- 18391460 TI - Preventive effect of lactoferrin intake on anemia in female long distance runners. AB - This study investigated whether intake of lactoferrin (LF) would improve or prevent anemia in female long distance runners who were training during the summer season and had a high risk of iron-deficiency anemia. Sixteen female long distance runners were divided into a group taking LF and iron (the LF group) and a group that only took iron (the control group) for 8 weeks. In the control group, the ferritin, serum iron, and red blood cell count were significantly lower than before treatment. In the LF group, the hematology data showed no significant change during the 8 weeks. The red blood cell count was significantly higher in the LF group than in the control group. The blood lactate level following a 3,000-m pace run of the control group was also significantly higher than that of the LF group. These observations suggest the possibility that intake of LF increases the absorption and utilization of iron and would be useful in the prevention of iron deficiency anemia among female long distance runners. PMID- 18391461 TI - Cloning, expression, and transglycosylation reaction of Paenibacillus sp. strain W-61 xylanase 1. AB - A xylanase gene, xyn1, which encodes Paenibacillus sp. strain W-61 xylanase 1 (Xyn1), was cloned in Escherichia coli. xyn1 encodes 211 amino acid residues, including 28 amino acid residues of a signal peptide. The deduced amino acid sequence of the mature Xyn1 showed 95.7%, 84.0%, and 83.7% identity to family 11 xylanases of Aeromonas caviae ME-1, Paenibacillus sp., and Bacillus stearothermophilus respectively. The physico-chemical properties of recombinant Xyn1 were very similar to those of intact Xyn1, except for the molecular mass. The pattern of xylooligosaccharides generated by rXyn1 was investigated by fluorophore-assisted carbohydrate electrophoresis (FACE). The degradation rate of xylohexaose by rXyn1 increased markedly as compared with that of xylopentaose. Xylohexaose had a single preferential point of cleavage by rXyn1. On the basis of the pattern of action of xylooligosaccharides, the number of subsites was estimated to be six. The catalytic site was located between the third and the fourth subsites from non-reducing end. PMID- 18391462 TI - A physiology study of Escherichia coli overexpressing phosphoenolpyruvate carboxykinase. AB - To determine whether intracellular ATP levels can be affected, Escherichia coli overexpressing phosphoenolpyruvate carboxykinase (pck) or phosphoenolpyruvate carboxylase (ppc) were grown in glucose minimal medium. The Pck-overexpressing cells showed approximately twice the intracellular ATP concentration, with 22% slower growth than the Ppc-overexpressing strain. This unexpected result of higher ATP coupled with slower growth is discussed based on transcriptome analysis. PMID- 18391463 TI - Heterologous expression of Na+/H+ antiporter gene (CvNHA1) from salt-tolerant yeast Candida versatilis in Saccharomyces cerevisiae Na+-transporter deficient mutants. AB - A Na(+)/H(+) antiporter gene (CvNHA1) was cloned from the salt-tolerant yeast Candida versatilis. CvNHA1 encodes an antiporter with a typical yeast plasma membrane Na(+)/H(+) antiporter structure. Transcription of CvNHA1 in C. versatilis cells was dependent on the salinity of the culture. When CvNHA1 was expressed in salt-sensitive Saccharomyces cerevisiae cells, increased salt tolerance was observed, indicating that Cvnha1p possesses an Na(+)/H(+) antiporter function, because the increased salt-tolerance was dependent on the extracellular pH. It appears that Cvnha1p mediates only the transport of Na(+). In an S. cerevisiae transformant harboring a CvNHA1-EGFP fusion plasmid in which the greater part of the C-terminal hydrophilic region of Cvnha1p was deleted by fusion with enhanced green fluorescent protein (EGFP), the Cvnha1-EGFP fusion protein was localized mainly in the plasma membrane, and the NaCl-tolerance of this transformant was greater than that of a strain harboring the entire CvNHA1 gene. PMID- 18391464 TI - Chiisanoside is not absorbed but inhibits oil absorption in the small intestine of rodents. AB - Chiisanoside is the main component of Acanthopanax sessiliflorus leaves. Simultaneous administration of chiisanoside resulted in a decrease in the plasma TG level and increase of undigested TG in the intestinal lumen after oil gavage to mice. This suggests that chiisanoside has the potential to prevent obesity as a lipase inhibitor which suppresses fat absorption in vivo. PMID- 18391465 TI - Identification of diterpene biosynthetic gene clusters and functional analysis of labdane-related diterpene cyclases in Phomopsis amygdali. AB - Two diterpene biosynthesis gene clusters in the fusicoccin-producing fungus, Phomopsis amygdali, were identified by genome walking from PaGGS1 and PaGGS4 which encode the geranylgeranyl diphosphate (GGDP) synthases. The diterpene cyclase-like genes, PaDC1 and PaDC2, were respectively located proximal to PaGGS1 and PaGGS4. The amino acid sequences of these two enzymes were similar to those of fungal labdane-related diterpene cyclases. Recombinant PaDC1 converted GGDP mainly into phyllocladan-16 alpha-ol via (+)-copalyl diphosphate (CDP) and trace amounts of several labdane-related hydrocarbons which had been identified from the P. amygdali F6 mycelia. Since phyllocladan-16 alpha-ol had not been identified in P. amygdali F6 mycelia, we isolated phyllocladan-16 alpha-ol from the mycelia. Recombinant PaDC2 converted GGDP into (+)-CDP. Furthermore, we isolated the novel diterpenoid, phyllocladan-11 alpha,16 alpha,18-triol, which is a possible metabolite of phyllocladan-16 alpha-ol in the mycelia. We propose that genome walking offers a useful strategy for the discovery of novel natural products in fungi. PMID- 18391466 TI - Low-molecular-weight hyaluronan permeates through human intestinal Caco-2 cell monolayers via the paracellular pathway. AB - The intestinal permeability of low-molecular-weight hyaluronan (LMW-HA) was investigated by using cultured monolayers of Caco-2 cells. The amount of LMW-HA that permeated the Caco-2 monolayers was measured by a carbazole assay. The permeability of LMW-HA increased inversely with the molecular size and was dose dependent. The transport was observed to be energy-independent, and was correlated with the tight junction (TJ) permeability. These results suggest that LMW-HA permeated the Caco-2 cell monolayers via the paracellular pathway. PMID- 18391467 TI - Cloning, purification, and polymerization of Capsicum annuum recombinant alpha and beta tubulin. AB - Alpha and beta tubulin genes were cloned from the Capsicum annuum leaves using rapid amplification of cDNA ends (RACE)-PCR. Nucleotide sequence analysis revealed that 1,353 bp Capsicum annuum alpha/beta-tubulin (CAnm alpha/beta-TUB) encodes a protein of 450 amino acids (aa) each. The recombinant alpha/beta tubulin was overexpressed mainly as an inclusion body in Escherichia coli BL21 (DE3), upon induction with 0.2 mM isopropyl-beta-D-thiogalactopyranoside (IPTG), and its content was as high as 50% of the total protein content. Effective fusion protein purification and refolding are described. The average yields of alpha and beta tubulin were 2.0 and 1.3 mg/l of culture respectively. The apparent molecular weight of each tubulin was estimated to be 55 kDa by SDS-polyacrylamide gel electrophoresis (PAGE). The tubulin monomers were found to be assembly competent using a standard dimerization assay, and also retained antigenicity with anti-His/T7 antibodies. The purified tubulins were polymerized to microtubule-like structures in the presence of 2 mM guanosine 5'-triphosphate (GTP). PMID- 18391468 TI - Oral administration of a hop water extract ameliorates the development of dermatitis induced by the periodical topical application of a mite antigen in atopic dermatitis model NC/Nga mice. AB - We investigated the inhibitory effect of an oral administration of a hop water extract (HWE) on the development of dermatitis by using NC/Nga atopic dermatitis model mice. The induction of allergic dermatitis was conducted by tape-stripping and topical application of a mite antigen (Dermatophagoides farinae) on to the ear once a week for 10 weeks. HWE was orally administered at a dose of 100 or 500 mg/kg. The total immunoglobulin E (IgE) concentration in serum and the ear thickness were periodically examined. Finally, the antigen-specific IgE level in the serum and the production of interleukin (IL)-4, IL-12 and interferon (IFN) gamma from splenocytes and cervical lymph node cells were measured. The oral administration of HWE significantly inhibited the increase of total IgE production and ear swelling throughout the experimental period. The production of IL-12 was significantly lower in the HWE administered group than in the control group. The results suggest that the intake of HWE may be effective in preventing and alleviating the development of atopic dermatitis-like skin disease. PMID- 18391469 TI - Potato and soy peptide diets modulate lipid metabolism in rats. AB - Dietary plant and animal peptides have been shown to reduce serum lipids. However, the potential of food-derived peptides has yet to be fully elucidated. We investigated the physiological importance of potato peptides in rats fed on a cholesterol-free diet containing 20% potato peptides (PP), when compared with two diets containing either 20% casein (CN) or 20% soy peptides (SP). The high density lipoprotein (HDL)-cholesterol (+13.8%) and serum triglyceride (-38%) concentrations in the PP-fed group, non-HDL-cholesterol level in the PP- (-22.5%) and SP- (-15.7%) fed groups, and serum total cholesterol concentration (-12%) in the SP-fed group, were significantly different from the control group at the end of the experiment. The fecal excretion of neutral and acidic sterols was higher in the PP- and SP-fed groups, respectively, relative to the control group. These results indicate that the observed changes in the serum cholesterol levels in rats fed on soy and potato peptide appear to have been due to different mechanisms. PMID- 18391470 TI - Isolation and characterization of Acidithiobacillus ferrooxidans strain D3-2 active in copper bioleaching from a copper mine in Chile. AB - Acidithiobacillus ferrooxidans strain D3-2, which has a high copper bioleaching activity, was isolated from a low-grade sulfide ore dump in Chile. The amounts of Cu(2+) solubilized from 1% chalcopyrite (CuFeS(2)) concentrate medium (pH 2.5) by A. ferrooxidans strains D3-2, D3-6, and ATCC 23270 and 33020 were 1360, 1080, 650, and 600 mg x l(-1) x 30 d(-1). The iron oxidase activities of D3-2, D3-6, and ATCC 23270 were 11.7, 13.2, and 27.9 microl O(2) uptake x mg protein(-1) x min(-1). In contrast, the sulfite oxidase activities of strains D3-2, D3-6, and ATCC 23270 were 5.8, 2.9, and 1.0 mul O(2) uptake.mg protein(-1).min(-1). Both of cell growth and Cu-bioleaching activity of strains D3-6 and ATCC 23270, but not, of D3-2, in the chalcopyrite concentrate medium were completely inhibited in the presence of 5 mM sodium bisulfite. The sulfite oxidase of strain D3-2 was much more resistant to sulfite ion than that of strain ATCC 23270. Since sulfite ion is a highly toxic intermediate produced during sulfur oxidation that strongly inhibits iron oxidase activity, these results confirm that strain D3-2, with a unique sulfite resistant-sulfite oxidase, was able to solubilize more copper from chalcopyrite than strain ATCC 23270, with a sulfite-sensitive sulfite oxidase. PMID- 18391471 TI - Enzymatic characterization of 5-methylthioribulose-1-phosphate dehydratase of the methionine salvage pathway in Bacillus subtilis. AB - 5-Methylthioribulose-1-phosphate (MTRu-1-P) dehydratase catalyzes the reaction from MTRu-1-P to 2,3-diketo-5-methylthiopentyl-1-phosphate (DK-MTP-1-P) in the methionine salvage pathway in Bacillus subtilis. The properties of this enzyme remain to be determined. We characterized these properties using a recombinant protein. The enzyme, with a molecular mass of 90 kDa, was composed of four subunits. The K(m) and V(max) of the enzyme were 8.9 microM and 42.7 micromole min(-1) mg protein(-1) at 25 degrees C respectively. Maximum activity was observed at pH 7.5 to 8.5 and 40 degrees C. The activation energy of the reaction from MTRu-1-P to DK-MTP-1-P was 63.5 kJ mol(-1). The reaction product DK-MTP-1-P was labile, and decomposed at a rate constant of 0.048 s(-1) to an unknown compound that was not utilized by DK-MTP-1-P enolase, the enzyme catalyzing the next step. The function of this enzyme in the pathway is discussed. PMID- 18391472 TI - Characterization of new 18-kDa IgE-binding proteins in beer. AB - The IgE-binding proteins in beer were examined by immunoblotting analysis with sera of patients sensitive to beer. Several proteins were immunoblotted with the sera, and among these, 18-kDa proteins were identified as new IgE-binding proteins in beer. Perhaps they originated from barley as a raw material. PMID- 18391473 TI - Low-density lipoprotein sub-fraction profiles in obese children before and after attending a residential weight loss intervention. AB - AIM: Small dense LDL particles are associated with an increased risk of coronary heart disease and are prevalent in obesity related dyslipidaemia. This study evaluated the effect of weight loss in nine children (BMI 33.4 +/- 8.4 kg.m(-2) and age 15.1 +/- 2.9 years) on LDL peak particle size, and cholesterol concentrations within particular LDL sub-fractions. METHODS: Each child undertook fun based physical activity, dietary restriction and modification and lifestyle education classes in a residential summer weight loss intervention. Blood was drawn before and after intervention and LDL heterogeneity measured by ultracentrifugation. RESULTS: The mean change in body weight were -6.8 +/- 4.9 kg, BMI units -2.5 +/- 1.4 kg.m(-2), and waist circumference -6.3 +/- 6.3 cm (all p < 0.01). Absolute LDL-c concentration reduced from 106.2 mg/dL to 88.3 mg/dL (p < 0.01). The cholesterol contained within the small dense LDL sub-fraction (LDL-c III) reduced from 54.1 mg/dL to 40.4 mg/dL (p < 0.01). Peak particle density decreased from 1.041g/mL to 1.035g/mL (p < 0.01). At pre intervention 50.9% of absolute cholesterol was within LDL-c III particles, changing to 46.2%. CONCLUSION: Mean weight loss of -6.8 +/- 4.9 kg lowers absolute LDL-c and the cholesterol specifically within LDL-c III particles. LDL peak particle size increased and a degree of LDL particle remodelling occurred. These favourable adaptations, accrued in a matter of 4 weeks, maybe associated with a reduction in CHD risk. PMID- 18391474 TI - Enzymatic synthesis of fatty hydrazides from palm oils. AB - Fatty hydrazides (FH) have been successfully synthesized from palm oils by a one step lipase catalyzed reaction. The synthesis was carried out by treating the oils with hydrazine hydrate at neutral pH using an immobilized lipase, Lipozyme as the catalyst. The percentages of conversion of RBD (refined, bleached and deodorized) palm oil (PO), RBD palm olein (POn), RBD palm stearin (PS) and RBD palm kernel olein (PKOn) into their fatty hydrazides are 95, 97, 97 and 99, respectively. PMID- 18391475 TI - High dose flaxseed oil supplementation may affect fasting blood serum glucose management in human type 2 diabetics. AB - Type 2 diabetes is characterized partially by elevated fasting blood serum glucose and insulin concentrations and the percentage of hemoglobin as HbA1c. It was hypothesized that each of blood glucose and its co-factors insulin and HbA1c and would show a more favorable profile as the result of flaxseed oil supplementation. Patients were recruited at random from a population pool responding to a recruitment advertisement in the local newspaper and 2 area physicians. Completing the trial were 10 flaxseed oil males, 8 flaxseed oil females, 8 safflower (placebo) oil males and 6 safflower oil females. Patients visited on two pre-treatment occasions each three months apart (visits 1 and 2). At visit 2 subjects were randomly assigned in double blind fashion and in equal gender numbers to take flaxseed oil or safflower oil for three further months until visit 3. Oil consumption in both groups was approximately 10 g/d. ALA intake in the intervention group was approximately 5.5 g/d. Power was 0.80 to see a difference of 1 mmol of glucose /L using 12 subjects per group with a p < 0.05. Flaxseed oil had no impact on fasting blood serum glucose, insulin or HbA1c levels. It is concluded that high doses of flaxseed oil have no effect on glycemic control in type 2 diabetics. PMID- 18391476 TI - Development of palm-based reference materials for the quantification of fatty acids composition. AB - Characterisation of fatty acids composition of three palm-based reference materials was carried out through inter-laboratory proficiency tests. Twelve laboratories collaborated in these tests and the fatty acids compositions of palm oil, palm olein and palm stearin were determined by applying the MPOB Test Methods p3.4:2004 and p3.5:2004. Determination of consensus values and their uncertainties were based on the acceptable statistical agreement of results obtained from the collaborating laboratories. The consensus values and uncertainties (%) for each palm oil reference material produced are listed as follows : 0.20% (C12:0), 1.66+/-0.05% (C14:0), 43.39+/-0.39% (C16:0), 0.14+/ 0.06% (C16:1), 3.90+/-0.11% (C18:0), 40.95+/-0.23% (C18:1), 9.68+/-0.21% (C18:2), 0.16+/-0.07% (C18:3) and 0.31+/-0.08% (C20:0) for fatty acids composition of palm oil; 0.23+/-0.04% (C12:0), 1.02+/-0.04% (C14:0), 39.66+/-0.19% (C16:0), 0.18+/ 0.07% (C16:1), 3.81+/-0.04% (C18:0), 44.01+/-0.08% (C18:1), 10.73+/-0.08% (C18:2), 0.20+/-0.06% (C18:3) and 0.34+/-0.04% (C20:0) for fatty acids composition of palm olein; and 0.20% (C12:0), 1.14+/-0.05% (C14:0), 49.42+/-0.25% (C16:0), 0.16+/-0.08% (C16:1), 4.15+/-0.10% (C18:0), 36.14+/-0.77% (C18:1), 7.95+/-0.29% (C18:2), 0.11+/-0.07% (C18:3) and 0.30+/-0.08% (C20:0) for fatty acids composition of palm stearin. PMID- 18391477 TI - Comparison of hydrogen bonding for chiral and racemic 1-monostearoylglycerols in solvent. AB - Comparison of the hydrogen bonding of racemic RS-1-monostearoylglycerol and chiral S-1-monostearoylglycerol in benzene was carried out through the NMR measurement. In addition, the concentration dependence of a chirality of S-1 monostearoylglycerol in hexane was studied though UV and circular dichroism (CD) measurements. The chemical shifts of OH protons of the S- and RS-1- monostearoylglycerols indicated that the hydrogen bonding between the R- and S form molecules of RS-1-monostearoylglycerol is stronger than that between the S- and S-form molecules of S-1-monostearoylglycerol in the low concentration region and that the difference in the strength of hydrogen-bonding between them becomes small in the high concentration region. The chirality of the S-1 monostearoyglycerol in hexane decreased with increasing its concentration. This suggests that the association of chiral S-1-monoacylglycerol arising from its increasing concentration reduces the chirality of S-1-monostearoylglycerol. PMID- 18391478 TI - Metabolism of odd-numbered fatty acids and even-numbered fatty acids in mouse. AB - Fatty acids are converted into energy via beta-oxidation. Although almost all natural occurring fatty acids are even-numbered, there are some odd-numbered fatty acids too. The details of the metabolism rate of odd-numbered fatty acids, however, are not clear. In the present study, we simultaneously administered a triacylglycerol containing four types of labeled even-numbered (palmitic acid and stearic acid) and odd-numbered (pentadecanoic acid and heptadecanoic acid) fatty acids to mice to compare the rates of their metabolism. The rates of metabolism were evaluated based on the accumulation of the labeled fatty acids in the small intestine epithelium, liver, and epididymal fat. Odd-numbered fatty acids accumulated mainly in the epididymal fat. In contrast, there was no accumulation of even-numbered fatty acids observed in the small intestine epithelium, liver, or epididymal fat. These results suggest that odd-numbered fatty acids might not be favorable substrates for beta-oxidation-related enzymes. PMID- 18391479 TI - Synthesis, surface-active properties, and antimicrobial activities of new double chain gemini surfactants. AB - A novel series of neutral and cationic dimeric surfactants were prepared involving ketalization reaction, Williamson etherification, and regioselective oxirane ring opening with primary and tertiary alkyl amines. The critical micelle concentration (CMC), effectiveness of surface tension reduction (gamma(CMC)), surface excess concentration (Gamma), and area per molecule at the interface (A) were determined and values indicate that the cationic series is characterized by good surface-active and self-aggregation properties. For the first time, we reported the antimicrobial activities against representative bacteria and fungi for dimeric compounds. The antimicrobial activity was found to be dependent on the target microorganism (Gram-positive bacteria > fungi > Gram-negative bacteria), as well as both the neutral or ionic nature (cationic > neutral) and alkyl chain length (di-C(12) > di-C(18) > di-C(8)) of the compounds. The cationic di-C(12) derivative was found to have equipotent activity to that of benzalkonium chloride (BAC) used as standard. PMID- 18391480 TI - Cholesterol-lowering activity of plant sterol-egg yolk lipoprotein complex in rats. AB - Free plant sterols cannot be dissolved in oil or water. Using free plant sterols and egg yolks, we developed a plant sterol-egg yolk lipoprotein complex (PSY) that can be dispersed in water and considered suitable for use in processed foods. The cholesterol-lowering activity of PSY was equal to that of free plant sterols and plant sterol esters. Consumption of a freeze-dried PSY-containing omelet reduced serum and hepatic cholesterol concentrations. The results suggest that PSY has cholesterol-lowering activity equivalent to that of free plant sterols and plant sterol esters. Moreover, the cholesterol-lowering activity of PSY was maintained in processed foods. PMID- 18391481 TI - Glucocorticoids ameliorate antigen-induced bronchial smooth muscle hyperresponsiveness by inhibiting upregulation of RhoA in rats. AB - To determine the mechanism(s) of the inhibitory effect of glucocorticoids on airway hyperresponsiveness in allergic bronchial asthma, the effects of systemic treatment with glucocorticoids on bronchial smooth muscle hyperresponsiveness and RhoA upregulation were investigated in rats with allergic bronchial asthma. Rats were sensitized and repeatedly challenged with 2,4-dinitrophenylated Ascaris suum antigen. Animals were also treated with prednisolone or beclomethasone (each 10 mg/kg, i.p.) once a day during the antigen inhalation period. Repeated antigen inhalation caused a marked bronchial smooth muscle hyperresponsiveness to acetylcholine with an upregulation of RhoA. Augmented acetylcholine-induced activation of RhoA and phosphorylation of myosin light chain were observed in bronchial smooth muscles of the antigen-exposed animals. Systemic treatment with either glucocorticoid used inhibited the bronchial smooth muscle hypercontraction until the level of the sensitized control rats that received saline inhalation instead of antigen challenge. Interestingly, both glucocorticoids also inhibited the upregulation of RhoA and augmented acetylcholine-induced activation of RhoA and phosphorylation of myosin light chain. In conclusion, glucocorticoids ameliorated the augmented bronchial smooth muscle contraction by inhibiting upregulation of RhoA. These effects of glucocorticoids may account for, in part, their beneficial effects in the treatment of asthma. PMID- 18391482 TI - Anti-inflammatory effect of theophylline in rats and its involvement of the glucocorticoid-glucocorticoid receptor system. AB - Although theophylline has been suggested to have an anti-inflammatory effect, there have been few reports to show the in vivo effect and the mechanism of anti inflammatory activity of theophylline experimentally. To reveal the anti inflammatory activity of theophylline, we studied the effect of theophylline and its metabolites on carrageenan-induced edema in rat foot pad. Subcutaneous injection of theophylline (5 - 100 mg/kg) inhibited carrageenan-induced edema dose-dependently. Theophylline metabolites, that is, 1-methylxanthine, 3 methylxanthine, 1-methyluric acid, and 1,3-dimethyluric acid (equimolar dose to 50 mg/kg of theophylline), did not inhibit the edema significantly. The inhibitory effect of theophylline on carrageenan-induced edema disappeared by pretreatment with aminoglutethimide, an inhibitor of glucocorticoid synthesis and with mifepristone, an antagonist of the glucocorticoid receptor. These results suggest that theophylline itself has anti-inflammatory activity and the glucocorticoid-glucocorticoid receptor system is involved in the anti inflammatory activity of theophylline. PMID- 18391483 TI - Peroxisome proliferator-activated receptor gamma (PPARgamma) suppresses colonic epithelial cell turnover and colon carcinogenesis through inhibition of the beta catenin/T cell factor (TCF) pathway. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma), a nuclear receptor superfamily member, plays a major role in lipid metabolism and insulin sensitivity. We investigated the role of PPARgamma in colonic epithelial cell turnover and carcinogenesis in colon because PPARgamma is strongly expressed in colonic epithelium. Administration of PPARgamma agonists suppressed epithelial cell turnover in mice. Expression level of beta-catenin protein, a key molecule in carcinogenesis, was increased in mouse colon treated with PPARgamma ligands. A direct interaction between beta-catenin and PPARgamma in cultured cell lines and colonic epithelium in mice was observed. Ligand-activated PPARgamma ligand directly interacts with beta-catenin, retaining it in the cytosol and reducing beta-catenin/T cell factor (TCF) transcriptional activity that is functionally important on aberrant crypt foci (ACF) formation. PPARgamma hetero-deficiency promoted the induction of ACF, but had no effect on the incidence of colonic polyps. These results indicate that PPARgamma regulates colonic epithelial cell turnover via direct interactions with beta-catenin, resulting in inhibition of beta-catenin-mediated transcriptional pathways that are involved in promoting cell proliferation. Our findings suggest that PPARgamma plays a role as a physiological regulator of colonic epithelial cell turnover and consequently predisposition to the development of colon cancer in early stage. PMID- 18391484 TI - Possible involvement of Akt activity in endothelial dysfunction in type 2 diabetic mice. AB - We investigated the effects of chronic simvastatin treatment on the impaired endothelium-dependent relaxation seen in aortas from type 2 diabetic mice. Starting at 8 weeks of diabetes, simvastatin (10 mg/kg per day) was administered to diabetic mice for 4 weeks. The significantly elevated systolic blood pressure in diabetic mice was normalized by simvastatin. Aortas from diabetic mice, but not those from simvastatin-treated diabetic mice, showed impaired endothelium dependent relaxation in response to both clonidine and adrenomedullin. After preincubation with an Akt inhibitor, these relaxations were not significantly different among the three Akt inhibitor-treated groups (controls, diabetics, and simvastatin-treated diabetics). Although clonidine-induced NO(x)(-) (NO(2)(-) + NO(3)(-)) production was greatly attenuated in our diabetic model, it was normalized by simvastatin treatment. The expression levels of both total Akt protein and clonidine-induced Ser-473-phosphorylated Akt were significantly decreased in diabetic aortas, while chronic simvastatin administration improved these decreased levels. The expression level of clonidine-induced phosphorylated PTEN (phosphatase and tensin homolog deleted on chromosome ten) was significantly increased in diabetic aortas, but chronic simvastatin did not affect it. These results strongly suggest that simvastatin improves the endothelial dysfunction seen in type 2 diabetic mice via increases in Akt and Akt phosphorylation. PMID- 18391485 TI - Catheter locking-solution spillage: theory and experimental verification. AB - A theory was developed allowing the prediction of the spillage volume after injection of locking solution into a catheter. This theory was corroborated by in vitro measurements using colorized fluid injected automatically by a micropump into a stirred container with clear fluid. The amount of spilled solution was measured with a fiberoptic spectrometer equipped with a dip probe. Straight cylindrical tubing was used as catheter. RESULTS: The theory predicts the beginning of spillage when 50% of the catheter filling volume is injected. When the nominal filling volume is injected, 25% of the solution is spilled and the mean concentration at the catheter tip is 50% of the locking solution concentration. EXPERIMENTS: Spillage was slightly less than predicted (20-25% at nominal filling volume). During the experiments it was recognized that a small air bubble trapped in the Luer connection during connection reduces the spillage volume to 10-15%. PMID- 18391486 TI - Donepezil treatment of patients with severe Alzheimer's disease in a Japanese population: results from a 24-week, double-blind, placebo-controlled, randomized trial. AB - BACKGROUND/AIMS: A 24-week, randomized, parallel-group, double-blind placebo controlled study was conducted to evaluate the efficacy and tolerability of donepezil in severe Alzheimer's disease (AD). METHODS: Patients with severe AD (Mini-Mental State Examination score 1-12; modified Hachinski Ischemic Score < or =6; Functional Assessment Staging > or =6) were enrolled in this study in Japan. A total of 325 patients were randomized to donepezil 5 mg/day (n = 110), donepezil 10 mg/day (n = 103) or placebo (n = 112). Primary outcome measures were change from baseline to endpoint in the Severe Impairment Battery (SIB) and Clinician's Interview-Based Impression of Change-plus caregiver input (CIBIC plus) at the endpoint visit. RESULTS: Donepezil 5 mg/day and 10 mg/day were significantly superior to placebo on the SIB, with a least-squares mean treatment difference of 6.7 and 9.0, respectively (p < 0.001 compared with placebo). CIBIC plus analyses showed significant differences in favor of donepezil 10 mg/day over placebo at endpoint (p = 0.003). A statistically significant dose-response relationship was demonstrated with the SIB and CIBIC-plus. Donepezil was well tolerated. CONCLUSION: This study confirmed the effectiveness of donepezil 10 mg/day in patients with severe AD and demonstrated a significant dose-response relationship. Donepezil at dosages of both 5 mg/day and 10 mg/day is safe and well tolerated in Japanese patients with severe AD. PMID- 18391487 TI - Treatment options in Alzheimer's disease: maximizing benefit, managing expectations. AB - Alzheimer's disease (AD) is becoming an increasingly heavy burden on the society of developed countries, and physicians now face the challenge of providing efficient treatment regimens to an ever-higher number of individuals affected by the disease. Currently approved anti-AD therapies - the cholinesterase inhibitors and the N-methyl-D-aspartate receptor antagonist memantine - offer modest symptomatic relief, which can be enhanced using combination therapy with both classes of drugs. Additionally, alternative therapies such as nonsteroidal anti inflammatory drugs, vitamin E, selegiline, Ginkgo biloba extracts, estrogens, and statins, as well as behavioral and lifestyle changes, have been explored as therapeutic options. Until a therapy is developed that can prevent or reverse the disease, the optimal goal for effective AD management is to develop a treatment regimen that will yield maximum benefits for individual patients across multiple domains, including cognition, daily functioning, and behavior, and to provide realistic expectations for patients and caregivers throughout the course of the disease. This review provides a basic overview of approved AD therapies, discusses some pharmacologic and nonpharmacologic treatment strategies that are currently being investigated, and offers suggestions for optimizing treatment to fit the needs of individual patients. PMID- 18391488 TI - Systematic appraisal using immunohistochemistry of brain pathology in aged and demented subjects. AB - BACKGROUND/AIMS: Abnormal processing of hyperphosphorylated tau (HPtau), amyloid beta (Abeta) and alpha-synuclein (alphaS) proteins is considered as causative with regard to the clinical symptoms in age-related neurodegenerative diseases. METHODS: In this retrospective, postmortem study applying immunohistochemical methodology, we assessed Alzheimer's-disease (AD)-related HPtau and Abeta pathology in 178 subjects with alphaS pathology. RESULTS: These pathologies were frequently seen concomitantly, i.e. HPtau in 83% and Abeta in 62% of the alphaS positive cases. Furthermore, the striatum was frequently involved, particularly in subjects with cognitive impairment (65%). The predictive value of widespread HPtau pathology, i.e. stages V-VI, with respect to cognitive impairment was high, since all 18 subjects presenting with this stage were demented. In contrast, the predictive value of widespread alphaS pathology, i.e. stages 5-6 according to Braak's Parkinson disease staging, was debatable. Fifty-three percent of the subjects with widespread alphaS pathology and no or mild AD-related HPtau pathology were cognitively unimpaired. It is noteworthy that striatal Abeta pathology was more often seen in demented subjects independently of HPtau and/or alphaS status. CONCLUSION: The causative pathology in subjects with clinically diagnosed dementia with Lewy bodies needs to be clarified in future studies. PMID- 18391489 TI - Functional voltage-gated sodium channels are expressed in human intestinal epithelial cells. AB - BACKGROUND AND AIM: Local anesthetics which preferentially interact with voltage gated sodium channels (VGSCs) were shown to have a clinical beneficial effect in ulcerative colitis and to regulate the secretion of inflammatory mediators from intestinal epithelial cells. However, expression of VGSCs in epithelial cells was not demonstrated. Herein we assessed whether intestinal epithelial cells express VGSCs. METHODS: The expression of VGSCs in normal human colonic tissue and in the TNFalpha-treated or untreated intestinal epithelial cell line HT-29 was studied by immunofluorescence staining and FACS analysis, Western blot, immunohistochemistry, and RT-PCR using primers specific for several VGSC alpha subunits. The function of VGSCs was assessed by measuring changes in the membrane potential of the intestinal epithelial cells following incubation with lidocaine, veratridine, or both. RESULTS: HT-29 cells were shown to express the VGSC alpha protein. mRNA analysis revealed the expression of nine of ten VGSC alpha isoforms. Immunohistochemical staining of normal colonic tissue confirmed the expression of VGSCs in colonic epithelial cells, smooth muscle cells, and nerves. Lidocaine induced membrane depolarization of HT-29 cells and its effect was blocked by veratridine. CONCLUSION: Malignant and normal intestinal epithelial cells express functional VGSCs. These molecules may play a role in the regulation of inflammation in gut physiology and pathology. PMID- 18391490 TI - Guiding manometry probes into the human small bowel without X-ray: validation of a new technique. AB - BACKGROUND: In order to perform small bowel manometry studies in man, the probes have to be guided through the upper gastrointestinal tract. This is usually controlled by fluoroscopy, but a method that allows to place the tubes without X rays is obviously desirable. METHODS: A new method that controls tube placing by sonography and pH-metry instead of X-rays is described. In a total of 19 volunteers 40 tubes were placed, either under fluoroscopic control (n = 22) or without X-rays (n = 18). RESULTS: Nose to antrum time was not significantly different between the two groups (8 +/- 2 vs. 10 +/- 2 min), but the documented transit of the tube from the antrum into the duodenum was significantly slower in the fluoroscopy group (80 +/- 20 vs. 60 +/- 12 min). Small bowel transit time was not significantly different between the groups (73 +/- 16 vs. 65 +/- 15 min). CONCLUSION: Placing motility probes in the human small bowel can be controlled by sonography and pH-metry instead of fluoroscopy. It is also easily performed, faster and more convenient for the investigator. As far as basic gastrointestinal research is concerned, in Germany less bureaucratic paperwork is involved, as these studies need not be approved by the 'Bundesamt fur Strahlenschutz' any more. PMID- 18391491 TI - Gastric antral vascular ectasia (GAVE): an update on clinical presentation, pathophysiology and treatment. AB - Gastric antral vascular ectasia (GAVE), though a rare disorder, causes up to 4% of non-variceal upper GI bleeding. This paper gives an overview of studies examining clinical presentation and pathophysiology, and reviews the current evidence for invasive and non-invasive treatments. GAVE is often associated with systemic illnesses, such as cirrhosis of the liver, autoimmune connective tissue disorders, bone marrow transplantation and chronic renal failure. The pathophysiological changes leading to GAVE have not been fully explained and remain controversial. Patient presentation varies from chronic iron-deficiency anaemia to heavy acute gastrointestinal bleeding. It is important to differentiate GAVE from portal hypertensive gastropathy as GAVE does not respond to measures reducing portal pressures. Endoscopic ablation (Nd:YAG-laser or argon plasma coagulation) is the first-line treatment of choice. As evidence for pharmacological therapy with oestrogen (and/or progesterone), tranexamic acid or thalidomide stems from case reports only, these should be used if endoscopic measures have failed to stop chronic blood loss. Surgical antrectomy should be reserved for unresponsive cases as it is associated with a high mortality. Ultimately, treatment of the underlying medical co-morbidities may lead to resolution of GAVE. PMID- 18391492 TI - Developmental anatomy of craniofacial sutures. AB - Sutures are fibrous joints in the vertebrate skull. They consist of two bone ends and intervening fibrous tissue which differentiates from embryonic mesenchyme. Sutures are not merely articulations between bones they are primary sites of osteogenesis mediating much of the growth of the face and skull vault. In this chapter the development of sutures will be described including the origin of sutural tissues, the determinants of suture location, and suture morphology. Also, the main functions of sutures will be explained. PMID- 18391493 TI - Locate, condense, differentiate, grow and confront: developmental mechanisms controlling intramembranous bone and suture formation and function. AB - The key mechanisms controlling where and when craniofacial bones and hence sutures form are discussed in this review. These include the formation and growth of skeletogenic condensations, tissue to tissue interactions between the epithelium, skeletogenic mesenchyme and the underlying dural and neural tissues. Also discussed are the key processes determining intramembranous bone growth, namely osteoblastogenesis and osteoclastogenesis. PMID- 18391495 TI - Suture neontology and paleontology: the bases for where, when and how boundaries between bones have been established and have evolved. AB - Much of what has been written about sutures has either focused on the genetic and biologic etiologies of specific sutural development, maintenance, and pathogenesis or on the utilization of sutures as character states in vertebrate cladistic analyses. There is a much more modest literature explicitly concerned with the evolution of sutures. We provide a small bridge of these literatures by presenting a discussion of the evolutionary biologic bases for the patterns of where, when, and how sutural boundaries between skeletal and dental elements have been established and have evolved. As sutural boundaries do not exist in the absence of the nucleation events that initiate the generation of skeletal elements, we explore historic models seeking to identify the inductive events dictating the specific times and places where a cranial skeletal element forms, the elaboration of its sutural boundaries, and the mechanisms whereby subsequent phyletic changes may be manifested and recognized. PMID- 18391494 TI - Mechanical influences on suture development and patency. AB - In addition to their role in skull growth, sutures are sites of flexibility between the more rigid bones. Depending on the suture, predominant loading during life may be either tensile or compressive. Loads are transmitted across sutures via collagenous fibers and a fluid-rich extracellular matrix and can be quasi static (growth of neighboring tissues) or intermittent (mastication). The mechanical properties of sutures, while always viscoelastic, are therefore quite different for tensile versus compressive loading. The morphology of individual sutures reflects the nature of local loading, evidently by a process of developmental adaptation. In vivo or ex vivo, sutural cells respond to tensile or cyclic loading by expressing markers of proliferation and differentiation, whereas compressive loading appears to favor osteogenesis. Braincase and facial sutures exhibit similar mechanical behavior and reactions despite their different natural environments. PMID- 18391496 TI - Single suture craniosynostosis: diagnosis and imaging. AB - Craniosynostosis, premature suture fusion, is one of the most common craniofacial anomalies affecting approximately 1 in 2,500 live births. Craniosynostosis is most commonly an isolated (nonsyndromic) condition with the sagittal suture being the most commonly affected suture. In this review we describe the range of isolated synostoses and show how these can lead to a variety of different morphological and functional abnormalities. PMID- 18391497 TI - Clinical features of syndromic craniosynostosis. AB - Disruption of normal suture development and function can result in premature suture fusion, craniosynostosis. This review focuses on syndromic forms of craniosynostosis. More than 100 syndromes in which craniosynostosis is a feature have been documented and here the most common conditions including Apert and Crouzon syndromes are described as well as other conditions with a particularly interesting molecular etiology, such as Saethre- Chotzen and craniofrontonasal syndrome. PMID- 18391498 TI - Genetics of craniosynostosis: genes, syndromes, mutations and genotype-phenotype correlations. AB - Craniosynostosis is a very heterogeneous group of disorders, in the etiology of which genetics play an important role. Chromosomal alterations are important causative mechanisms of the syndromic forms of craniosynostosis accounting for at least 10% of the cases. Mutations in 7 genes are unequivocally associated with mendelian forms of syndromic craniosynostosis: FGFR1, FGFR2, FGFR3, TWIST1, EFNB1, MSX2 and RAB23. Mutations in 4 other genes, FBN1, POR, TGFBR1 and TGFBR2, are also associated with craniosynostosis, but not causing the major clinical feature of the phenotype or with an apparently low penetrance. The identification of these genes represented a great advance in the dissection of the genetics of craniosynostosis in the last 15 years, and today they explain the etiology of about 30% of the syndromic cases. The paucity in the identification of genes associated with this defect has partly been due to the rarity of familial cases. In contrast, very little is known about the molecular and cellular factors leading to nonsyndromic forms of craniosynostosis. Revealing the molecular pathology of craniosynostosis is also of great value for diagnosis, prognosis and genetic counseling. This chapter will review (1) the chromosomal regions associated with syndromic forms of the malformation, (2) the genes in which a large number of mutations have been reported by independent studies (FGFR1, FGFR2, FGFR3, TWIST1 and EFNB1) and (3) the molecular mechanisms and genotype phenotype correlations of such mutations. PMID- 18391499 TI - Roles of FGFR2 and twist in human craniosynostosis: insights from genetic mutations in cranial osteoblasts. AB - Recent advances in molecular genetics have led to a better understanding of the role of specific genes such as fibroblast growth factor receptor (FGFR) and Twist in cranial bone formation. Specifically, the analysis of osteoblast abnormalities induced by FGFR2 and Twist genetic mutations inducing craniosynostosis in humans has provided some insights into the role of these genes in the premature cranial suture formation in syndromic craniosynostosis. This also led to a better understanding of the cellular and molecular mechanisms that control osteoblast biology and pathology in humans. In this review paper, we summarize the effects of FGFR2 and Twist genetic mutations resulting in altered osteoblast phenotype and premature cranial fusion based on our analysis in human syndromic craniosynostosis. PMID- 18391500 TI - Fibroblast growth factor signaling in cranial suture development and pathogenesis. AB - Apert, Pfeiffer and Crouzon syndromes are congenital craniosynostosis syndromes caused by mutations that perturb the level of fibroblast growth factor receptor (FGFR) signaling. The cellular and molecular impact of these mutations have been studied in vitro and in animal models in vivo. Here, I highlight the complexity of the FGF/FGFR signaling system and review the candidate modifiers responsible for regulating the levels of FGF/FGFR signaling in tissues. I also review what we have learned from the phenotypic analysis of mice that model these craniosynostosis syndromes and discuss some in vivo strategies for further understanding as well as alleviating the associated craniofacial defects. PMID- 18391501 TI - Tgf-beta regulation of suture morphogenesis and growth. AB - Premature suture obliteration results in an inability of cranial and facial bones to grow, with resulting craniofacial dysmorphology requiring surgical correction. Understanding the biological signaling associated with suture morphogenesis will enable less invasive treatment of patients with fused sutures, combined with therapy using biological molecules. While a number of advances have been made in identifying the genetic etiologies of various craniosynostotic syndromes, the pathogenesis of this condition is still not completely understood. Recently, it has been shown that differential expression of various transforming growth factor beta (Tgf-beta) isoforms plays a crucial role in regulating suture patency once the sutures have formed. It has also been shown that differential expression of Tgf-beta isoforms may also play a role in craniosynostosis by altering proliferation, differentiation, and apoptosis within the suture. This chapter focuses on the role of Tgf-beta in suture morphogenesis and growth, exploring Tgf beta biology, receptors, signaling pathways, animal models, and expression in both normal and pathological sutures. PMID- 18391502 TI - The Bmp pathway in skull vault development. AB - The Bmp pathway is of critical importance in the development of the skull vault. Analysis of gain and loss of function phenotypes of Bmp pathway effectors, particularly Msx genes, has shown that the Bmp pathway functions in the growth of both mesodermal and neural crest-derived calvarial bones. It is required for the development of the frontal and parietal bones during the interval between the initial osteogenic mesenchymal condensations at E12.5 to the apposition of the paired frontal and parietal bones at E18.5. During postnatal development, forced expression of the Bmp inhibitor, noggin, maintains the patency of sutures, consistent with a role for the Bmp pathway in regulating suture development. The availability of conditional mutants of Bmp ligands, receptors and downstream effectors will make possible an increasingly high resolution analysis of precisely how the Bmp functions in these processes and how aberrations in its activity can contribute to pathological conditions such as familial parietal foramina and craniosynostosis. PMID- 18391503 TI - Current treatment of craniosynostosis and future therapeutic directions. AB - Normal craniofacial development is contingent upon coordinated growth between the brain and overlying calvaria. Craniosynostosis, the premature fusion of one or more cranial sutures, perturbs this natural framework, resulting in dramatic dysmorphology of the skull and face along with a multitude of associated functional abnormalities. Traditional approaches to the treatment of craniosynostosis have employed complex surgical remodeling of the skull vault and facial deformities all aimed at increasing the amount of intracranial volume and restoring a more normal craniofacial appearance. Significant morbidity and mortality, however, have plagued these procedures, driving dramatic evolution in our approach towards the treatment of pathologically fused sutures. Recent clinical and genetic studies have identified multiple forms of human craniosynostosis, each associated with mutations within various cytokine signaling pathways. Knowledge garnered from these investigations bear promise for the future development of alternative strategies to enhance or perhaps even replace contemporary approaches for the treatment of craniosynostosis. PMID- 18391504 TI - Capsaicin inhibits proliferation of endometriotic cells in vitro. AB - Treatment of immortalized stromal-like and epithelial-like endometriotic cells with capsaicin resulted in inhibition of proliferation in a concentration dependent manner. In addition, endometriotic cells are more sensitive to capsaicin treatment than immortalized endometrial cells, suggesting that capsaicin may be an admissible drug candidate for treating endometriosis. PMID- 18391505 TI - Podocyte injury induced by albumin overload in vivo and in vitro: involvement of TGF-beta and p38 MAPK. AB - BACKGROUND: Proteinuria is a well-established exacerbating factor in chronic kidney disease. Although the mechanisms of albumin-induced tubulointerstitial damage have been extensively studied, the influence of proteinuria on podocytes has not been sufficiently elucidated. The present study examined the effect of albumin overload on podocytes in vivo and in vitro and explored the underlying mechanisms. METHODS: Rat podocytes were exposed to albumin overload in vivo by the intraperitoneal injection of albumin over 2 days whilst cultured podocytes were subjected to albumin in vitro. We analyzed albumin uptake, podocyte apoptosis, staining for F-actin and nephrin and involvement of TGF-beta1 and p38 MAPK cascades. RESULTS: Rats administered albumin exhibited massive proteinuria and podocyte injury manifested by decreased nephrin immunostaining and foot process effacement. These abnormalities were accompanied by albumin deposition, TGF-beta1 upregulation, p38 MAPK phosphorylation and an increased number of glomerular TUNEL-positive cells. Exposure of cultured podocytes to albumin caused actin disarrangement and apoptosis. Podocyte injury was preceded by albumin uptake, induction of TGF-beta1 and phosphorylated p38 MAPK. Treatment of podocytes with anti-TGF-beta1 neutralizing antibody or SB203580 significantly reduced the albumin-induced injury. CONCLUSIONS: These results indicate that albumin overload in vivo and in vitro promotes podocyte injury mainly via TGF beta1/p38 MAPK pathways. PMID- 18391507 TI - The anthropological role of the doctor: do our patients feel better? PMID- 18391506 TI - Chemokine MIP-2/CXCL2, acting on CXCR2, induces motor neuron death in primary cultures. AB - OBJECTIVES: Chemokines are implicated in many diseases of the central nervous system (CNS). Although their primary role is to induce inflammation through the recruitment of leukocytes by their chemotactic activity, they may also have direct effects on neuronal cells. We evaluated the expression of CXCR1 and CXCR2 and investigated the effect of CXCR2 activation by the agonist MIP-2 (CXCL2) on primary cultured motor neurons. To specifically assess the role of CXCR2 in the neurotoxicity induced by MIP-2, we used the CXCR1/2 inhibitor reparixin and studied the effect of the chemokine on motor neuron cultures from CXCR2-deficient mice. METHODS: Primary motor neurons prepared from rat or mouse embryos were treated with MIP-2 and reparixin. Motor neuron viability and receptor expression were assessed by immunocytochemical techniques. RESULTS: Rat primary motor neurons expressed CXCR2 receptors and recombinant rat MIP-2 induced dose dependent neurotoxicity. This neurotoxicity was counteracted by reparixin, a specific CXCR1/2 inhibitor, and was not observed in motor neurons from CXCR2 deficient mice. CONCLUSIONS: CXCR2 activation might directly contribute to motor neuron degeneration. Thus, chemokines acting on CXCR2, including IL-8, may have direct pathogenic effects in CNS diseases, independent of the induction of leukocyte migration. PMID- 18391508 TI - Morphine-induced behavioral sensitization increased the mRNA expression of NMDA receptor subunits in the rat amygdala. AB - This study was designed to evaluate the effect of repeated morphine treatment on rat behavioral responses. In the genetic section, the mRNA expression of NMDA receptor subunits (NR1 and NR2A) was measured in certain areas of the male rat brain (striatum, prefrontal cortex, hippocampus, hypothalamus and amygdala). In the behavioral section, the effect of repeated morphine treatment on animal models such as locomotion, oral stereotypy, and state-dependent memory in a passive avoidance test was evaluated in the presence or absence of MK801 (NMDA receptor antagonist). Our results showed that chronic morphine treatment, followed by a 7-day (but not 24-hour) washout period, potentiated the effect of test doses of morphine, which is referred to as behavioral sensitization. Meanwhile, pretreatment of animals with MK801 (0.1 and 0.25 mg/kg), 30 min before a test dose of morphine (5 mg/kg), failed to attenuate the locomotion and oral stereotypy in the behavioral sensitization state. Interestingly, a higher dose of MK801 (0.25 mg/kg) decreased memory retrieval induced by morphine (2.5 mg/kg) in state-dependent memory. This effect may be due to the intrinsic motor enhancer property of higher doses of MK801, rather than the blockade of NMDA receptors. It can be concluded that MK801 does not affect morphine-induced behavioral sensitization in the expression phase. In the genetic section of the study, results of quantitative real-time RT-PCR clearly indicated that morphine sensitization increased the expression of NMDA receptor subunits mRNA in the amygdala (NR1 by 104% and NR2A by 85%), while the other areas of the brain were unaffected. Maenwhile, no change in the mRNA levels was observed in non sensitized animals (chronic morphine treatment followed by a 24-hour washout period). In summary, the present study indicates that repeated morphine treatment followed by long-term (7-day washout) induces behavioral sensitization and causes a delayed increase in mRNA levels of NMDA receptor subunits in the rat amygdala. Meanwhile, it has previously been reported that the amygdala is involved in behavioral sensitization. Thus, it can be concluded that the increase in NMDA receptor expression is associated with morphine-induced behavioral sensitization. PMID- 18391509 TI - Pharmacokinetics of isoflurane in human blood. AB - Investigation of isoflurane washout from the human body and brain provides more precise information about elimination in anesthesia. The elimination pattern of isoflurane remains poorly quantified, and therefore this study tried to clarify the pharmacokinetic pattern of isoflurane elimination. Sixteen patients (aged 48 78 years), undergoing coronary arterial bypass grafting surgery were enrolled in this study. Sixty minutes prior to the end of surgery, we kept a fixed 2% inspired isoflurane in 6,000 ml min(-1) oxygen flow. Isoflurane supplement was then discontinued to study the 20-min isoflurane elimination. An infrared analyzer was used to determine both inspired isoflurane and end-tidal isoflurane. The isoflurane concentration in the internal jugular bulb blood, arterial blood and pulmonary arterial blood were analyzed by gas chromatography. Biexponential decay function was the best fitted for the end-tidal isoflurane- and arterial blood isoflurane-time curves. There were two distinct components, including initial 5-min fast component and the next 15-min slow component. Monoexponential decay function was the best fitted for the pulmonary arterial blood- and jugular bulb blood-time curves. During elimination, the initial washout of isoflurane from functional residual capacity of lungs is reflected in the fast component of the isoflurane concentration time curves. The later slow component is dominated by the tangible manifestation of physiological membrane barriers, including the existence of alveoli-pulmonary capillary, blood-brain barriers. PMID- 18391510 TI - Single nucleotide polymorphisms of the pregnane x receptor gene in Han Chinese and a comparison with other ethnic populations. AB - The pregnane X receptor (PXR/NR1I2) gene is a master regulator for a number of cytochrome P450s (CYPs) and drug transporters. This study aimed to detect the single nucleotide polymorphisms (SNPs) of the PXR gene in Han Chinese (n = 186) and to compare the frequencies of polymorphisms of the PXR gene with those in Caucasian and African Americans reported in the literature. The SNPs of the PXR gene were analyzed using the polymerase chain reaction (PCR) and direct sequencing analysis. The mutant frequencies of A11156C and T11193C in Han Chinese were 55% (95% confidence interval (CI): 0.49-0.61) and 59% (95% CI: 0.52-0.64), respectively, higher than those of Caucasian Americans (16 and 16%, respectively) and African Americans (33 and 30%, respectively). However, the reported SNPs in exons 2 and 4 (PXR*2,*3,*4,*6,*9,*10,and *11) were not detected in Han Chinese. These results indicate that there are marked differences in the mutant frequencies of A11156C and T11193C of PXR between Han Chinese and other ethnic groups. The mutant frequency in the coding region (exons 2 and 4) of PXR was very low in Han Chinese. Further studies are needed to determine the impact of common SNPs of PXR in Han Chinese and other ethnic populations on the phenotypic activity of cytochrome P450s and drug transporters transactivated by PXR. PMID- 18391511 TI - The evolution of sex chromosomes and sex determination in vertebrates and the key role of DMRT1. AB - Mechanisms of vertebrate sex determination are reviewed in the light of what is known about the origin and evolution of sex chromosomes. All vertebrate males have testes that are similar in anatomy and in spermatogenesis. Despite a variety of sex chromosome systems, the same cascade of genes operate in the differentiation of testes and male genitalia. Only the primary signal or switch that initiates the cascade may be different. Genetic sex determination (GSD) occurs in most species, although temperature-dependent sex determination (TSD) is the switch in some reptiles without GSD. SRY is the genetic switch in eutherians (placental mammals) and DMRT1 may have that function in birds and at least one species of fish. The switch in all other groups of vertebrates with either male or female heterogamety is unknown. DMRT1 is an ancient sex determining gene, found first in invertebrates, and is one of several genes expressed in higher levels in the vertebrate embryonic testis than in the embryonic ovary. It is singled out in this review because of its likely key role in sex determination both in birds and in reptiles with TSD. DMRT1 is one of the few sex genes that have been mapped in representative species of fish, turtles, crocodiles, amphibians, birds and mammals. Greater effort in comparative mapping, sequencing and expression studies is required to discover the other primary switch genes in vertebrates and to answer questions about their evolution. PMID- 18391512 TI - Involvement of homeobox genes in mammalian sexual development. AB - In eutherian mammals, sexual development hinges on the differentiation of the gonadal primordia into either testes or ovaries, in turn determined by the activity of the Y-linked gene Sry. Organogenesis of the gonads requires input from a complex network of morphogenetic signals, many of which are still uncharacterised. Homeobox genes are known to play important roles in embryo morphogenesis and organogenesis in a number of systems. Here, we review current data relating to possible roles of the extended homeobox gene family on gonad development, sex-specific morphogenesis and gametogenesis, and speculate that these genes may play a broader role in these processes than is currently suspected. PMID- 18391513 TI - Extended pedigree with multiple cases of XX sex reversal in the absence of SRY and of a mutation at the SOX9 locus. AB - It is well established that testicular differentiation of the human embryonic gonad depends on the action of the Y-chromosomal gene SRY. However, exceptional cases such as SRY-negative cases of 46,XX testicular disorder of sexual development (DSD), and of 46,XX ovotesticular DSD document that testicular tissue can develop in the absence of the SRY gene. These SRY-negative XX sex reversal cases are very rare and usually sporadic, but a few familial cases have been reported. We present a large, consanguineous family with nine affected individuals with phenotypes ranging from 46,XX testicular DSD to 46,XX ovotesticular DSD, with predominance of male characteristics. Absence of SRY in peripheral blood was documented by fluorescence in situ hybridization (FISH) and PCR analysis in all nine affected individuals, and by FISH analysis on gonadal sections with testicular tissue in four affected individuals. By quantitative PCR, a duplication of the SOX9 gene was excluded. In addition, as linkage analysis showed that the nine affected members of the family do not share a common SOX9 haplotype, any mutation at the SOX9 locus could be ruled out. Together, these findings implicate a mutation at a sex-determining locus other than SRY and SOX9 as the cause for the XX sex reversal trait in this family. PMID- 18391514 TI - Familial translocation t(1;9) associated with macromastia: molecular cloning of the breakpoints. AB - A familial reciprocal translocation associated with severe macromastia has been characterized by molecular cytogenetic and molecular analysis. Cloning of the translocation breakpoints revealed that no known gene has been disrupted by this translocation. Therefore, a position effect compromising the regulation of a still to be identified gene in the vicinity of the breakpoints can be assumed. PMID- 18391515 TI - mRNA expression analysis and the molecular basis of neonatal testis defects in Dmrt1 mutant mice. AB - Transcriptional regulators containing the DM domain DNA binding motif have been found to control sexual differentiation in a diverse group of metazoan animals including vertebrates, insects, and nematodes, suggesting that these proteins may comprise a very ancient group of sexual regulators. Dmrt1, 1 of 7 mammalian DM domain genes, is essential for several aspects of testicular differentiation in mice. The Dmrt1 mutant phenotype becomes apparent shortly after birth, and culminates in severe testicular dysgenesis. To better understand the roles of Dmrt1 in testicular development we have performed a more detailed analysis of its mutant phenotypes, and we have used mRNA expression profiling to identify genes misregulated in the neonatal Dmrt1 mutant testis. We find that Dmrt1 mutant germ cells fail to undergo several of the normal postnatal events of germ cell development, including radial movement, mitotic proliferation, differentiation into spermatogonia, and initiation of meiosis, and they die by P14. During this period Dmrt1 mutant Sertoli cells fail to polarize and form tight junctions, and fail to cease proliferation, eventually filling the seminiferous tubules. Expression profiling at P1 and P2 in Dmrt1 mutant testes indicates defects in several important testicular signaling pathways (Gdnf, retinoic acid, TGFbeta, FSH), and detects elevated expression of the pluripotency marker Stella/Dppa3/Pgc7, providing insight into the molecular basis of Dmrt1 testis defects. This work also identifies a number of new candidate testicular regulators for further investigation. PMID- 18391516 TI - A diploid-triploid (60,XX/90,XXY) intersex in a Holstein heifer. AB - The diploid/triploid (60,XX/90,XXY) condition in Bos taurus is very rare and only three cases have been published previously. The present animal exhibited an aplastic vulva, penis and clitoris agenesis, a male-like urethra located in a pseudoprepuce opening between the mammary complexes and a well developed M. rectipeninus. A normal (60,XX) female karyotype was detected in lymphocyte cultures whereas uterus and tendon cells revealed a 60,XX/90,XXY mixoploidy. Quantification of X and Y chromosome-specific sequences using RT-PCR revealed extraordinary high Y chromosome equivalents in the sample recovered from the male like transformed vestibulum vaginae suggesting a causative relationship. The pathogenesis of the missing clitoris and penis, which is contrasted by the concomitant presence of a well developed M. rectipeninus, remains difficult to explain. A chimeric origin is suggested despite the fact that microsatellite analysis of the animal's blood cells displayed no un- usual allele accumulation. PMID- 18391517 TI - Meiosis onset is postponed to postnatal stages during ovotestis development in female moles. AB - In mammals, germ cells are important both during development and for the function of female gonads, whereas male gonads may develop in the absence of germ cells. The gonads of female moles (genus Talpa) develop according to a testis-like pattern which results in the formation of ovotestes. In this paper, we studied the expression pattern of several pre-meiotic and meiotic germ cell markers, in order to establish the precise time of meiosis onset in the mole species T. occidentalis, and to investigate the location and possible role of germ cells in ovotestis organogenesis. Our results evidenced that: (1) the asymmetrical distribution of primordial germ cells, which concentrate in the cortex of the XX gonad, is brought about by germ cell depletion from the medulla between the s5a and s5b stages, (2) XX germ cells enter meiosis postnatally, which is quite exceptional among eutherian mammals, and (3) XX but not XY germ cells of moles express DMRT1 during premeiotic stages of development, an expression pattern not described previously in vertebrates. PMID- 18391518 TI - Testosterone stimulates mounting behavior and arginine vasotocin expression in the brain of both sexual and unisexual whiptail lizards. AB - In nonmammalian vertebrates the abundance of arginine vasotocin (AVT) neurons in the brain is sexually dimorphic, a pattern that is modulated by testicular androgen. This peptide is thought to be involved in the control of male-typical mounting behaviors. The all-female desert-grasslands whiptail (Cnemidophorus uniparens) reproduces by obligate parthenogenesis and in nature no males exist, but eggs treated with aromatase inhibitor hatch into individuals (called virago C. uniparens) having testes, accessory sex structures, high circulating concentrations of androgens, and exhibiting only male-like copulatory behavior. To examine the 'sexual' dimorphism of AVT-containing neurons in these animals, we compared AVT immunoreactivity in gonadectomized control and virago C. uniparens, with that of gonadectomized male and female Cnemidophorus inornatus, a sexual species that is the maternal ancestor to the parthenogenetic species. Mounting behavior is elicited in both species and both sexes by testosterone, and it was predicted that the distribution and abundance of AVT cell bodies and fibers would reflect the propensity of males and females of the two species to display male typical copulatory behavior. Since both this propensity and AVT abundance are controlled by androgens, we compared testosterone-implanted and control animals within each group. Testosterone treatment generally increased AVT abundance, except in lab-reared parthenoforms, in which testosterone treatment was the least effective in inducing male-like copulatory behavior. PMID- 18391519 TI - Governing sex determination in fish: regulatory putsches and ephemeral dictators. AB - In contrast to the rather stable regulatory regimes established over more that 100 million years in birds and mammals, sex determination in fish might frequently undergo evolutionary changes bringing the sex-determining cascade under new master sex regulators. This phenomenon, possibly associated with the emergence of new sex chromosomes from autosomes, would explain the frequent switching between sex determination systems observed in fish. In the medaka Oryzias latipes, the Y-specific master sex-determining gene dmrt1bY has been formed through duplication of the autosomal gene dmrt1 onto another autosome, thus generating a new Y chromosome. Dmrt1bY emerged about 10 million years ago and is restricted to several Oryzias species, indicating that the Y chromosome of the medaka is evolutionarily much younger than mammalian and bird sex chromosomes. Fertile males without dmrt1bY have been detected in some medaka populations, and this gene might even have been inactivated in one Oryzias species, indicating the existence of sexual regulators already able to supplant dmrt1bY. Studies on other models have confirmed that fish sex chromosomes are generally young and occurred independently in different fish lineages. The identification of new sex-determining genes in these species will shed new light on the exceptional evolutionary instability governing sex determination in fish. PMID- 18391520 TI - Helper function of the Wolffian ducts and role of androgens in the development of the vagina. AB - The development of a vagina as a separate outlet of the birth canal evolves at the transition of egg laying species to eutherian mammals. The derivation of the vagina from the Wolffian and Mullerian ducts and the contribution of the urogenital sinus are still open questions. Here experiments with the complete androgen receptor defect in the testicular feminisation (Tfm) mouse are reported which show that the vagina is formed by caudal migration of Wolffian and Mullerian ducts. The cranial ends of the Wolffian ducts successively regress while the Mullerian ducts fuse to form the vagina. Immunohistochemistry of the androgen receptor reveals that the caudal ends of the Wolffian ducts remain in the indifferent stage and therefore have been mistaken as sinuvaginal bulbs. The Wolffian ducts do not contribute to the vagina itself but have a helper function during downward movement of the vaginal bud in the female. In the male the caudal ends serve as androgen operated switch for the negative control of vaginal development. The results indicate that the rudimentary vagina in the complete androgen insensitivity syndrome (CAIS) corresponds to non obliterated caudal ends of the Mullerian ducts. Selective atresia of the vagina in the MRKH (Mayer Rokitansky-Kuster-Hauser) syndrome may be explained by the failure of Wolffian and Mullerian ducts to descend caudally. PMID- 18391521 TI - WNT4 and RSPO1 are not involved in a case of male-to-female sex reversal with partial duplication of 1p. AB - Some studies suggest that WNT4 signaling acts to repress the male pathway and a single case of duplication of WNT4 located in 1p35 has been associated with ambiguous genitalia. Recently, RSPO1 located in 1p34.3 was shown to be essential in sex determination. We show that neither WNT4 nor RSPO1 are duplicated in another case of male-to-female sex reversal with partial duplication of 1p. Therefore, it can be assumed that in male-to-female sex reversal associated with duplication of 1p, other genetic factors may be involved. PMID- 18391522 TI - Characterisation of urogenital ridge gene expression in the human embryonal carcinoma cell line NT2/D1. AB - The study of the mammalian sex-determining pathway has been hampered by the lack of cell culture systems to investigate the underlying molecular relationships between sex-determining genes. Recent approaches using high-throughput genome wide studies have revealed a number of sexually dimorphic genes expressed in the developing mouse gonad. Here, we investigated a human testicular cell line in terms of its expression of known sex-determining genes and newly identified candidates. The human embryonal carcinoma cell line NT2/D1 was screened for the expression of 46 genes with known or potential roles in the sex-determining and differentiation pathway. Forty genes tested were expressed in NT2/D1 cells including the testis-determining genes SRY, SOX9, SF-1, DHH and FGF9. Genes not expressed included WT1, DAX1 and the ovary-specific genes FOXL2 and WNT4. Cell specific markers demonstrate that NT2/D1 cells reflect a number of cell types in the gonad including Sertoli, Leydig and germ cells. Our results suggest that male pathways initiated by SRY, SOX9 and SF-1 remain intact in these cells. Lack of expression of ovary-specific genes is consistent with a commitment of these cells to the male lineage. Manipulation of gene expression in this cell line could be an important new in vitro tool for the discovery of new human sex-determining genes. PMID- 18391523 TI - Bmp7 regulates germ cell proliferation in mouse fetal gonads. AB - Relatively little is known regarding the signals that regulate the proliferation and sex-specific development of germ cells during mammalian fetal gonad differentiation. Members of the bone morphogenetic protein (BMP) family have been identified as key regulators of germ cells in the Drosophila gonad. Here we show that in mice Bmp7 is expressed in gonads of both sexes and is required for germ cell proliferation during a narrow window of development between 10.5-11.5 days post coitum (dpc). The proliferation defect is more severe in male than in female embryos suggesting that there are sexually dimorphic compensatory pathways. BMP signaling appears to be an evolutionarily conserved pathway regulating embryonic germ cell proliferation in vertebrate and invertebrate species. PMID- 18391524 TI - Temperature-dependent sex determination in Hd-rR medaka Oryzias latipes: gender sensitivity, thermal threshold, critical period, and DMRT1 expression profile. AB - The developmental time and thermal threshold for temperature-dependent sex determination (TSD), gender differences in temperature sensitivity, the fertility of thermally sex reversed fish, and the effect of temperature on the expression of two major sex determination/differentiation genes (DMY/DMRT1bY and DMRT1) were examined in the Hd-rR strain of medaka, Oryzias latipes. Fertilized eggs were exposed from either shortly after fertilization (8-16 cells; embryonic stages 5 6) or from middle embryogenesis (heart development stage; stage 36) until hatching to temperatures ranging from 17 degrees C to 34 degrees C. Secondary sexual characteristics, gonadal histology, progeny testing, sex-linked body coloration and gene expression were used to determine phenotypic and genotypic sex. Sex determination was unaffected by low or high temperatures in genotypic (XY) males. In contrast, genotypic (XX) females treated from stages 5-6 showed increasing rates of sex reversal into phenotypic males at temperatures above 27 degrees C up to 100% at 34 degrees C. Thermal manipulation of sex was ineffective after stage 36, indicating that gonadal fate in medaka is determined considerably earlier than histological differentiation (stage 39). High temperature induced DMRT1 expression in genotypic females, which was observed already from stage 36. Sex-reversed males had histologically normal testes, were capable of sexual courtship and, with the exception of fish from 34 degrees C, sired viable progeny when mating with fertile females. These results clarify the pattern of TSD in medaka and provide important clues to understand the mechanism of sex determination in this species. They also suggest that a brief exposure to high temperature early in life could impair the fertility of medaka as adults. PMID- 18391525 TI - Early diagnosis of 5alpha-reductase deficiency in newborns. AB - 5Alpha-reductase-2 deficiency is a rare autosomal recessive form of 46,XY disorders of sex differentiation (DSD), caused by mutations in the steroid 5alpha reductase type 2 gene (SRD5A2), presenting at birth with variable degrees of undervirilization. We report on three Italian newborns with 46,XY DSD in whom the evaluation of testosterone, dihydrotestosterone, testosterone/dihydrotestosterone (T/DHT) ratio and molecular analysis of the 5alpha-reductase type 2 gene was made in their first month of life. Baseline T/DHT ratio suggested 5alpha-reductase-2 deficiency; the diagnosis was confirmed by molecular genetics (homozygous mutation in exon 4 [G196S], heterozygous mutation in exon 1 and 5 [W35X/Y235F], heterozygous mutation plus polymorphism in exon 1 [G34W/A49T]). Proper investigation permitted early reassignment to male sex in two babies, assigned to female sex just after birth. In infancy, the T/DHT ratio, assessed by suitable assay methods and evaluated by age-appropriate reference values, seems to be able to select newborns affected by 5alpha-reductase-2 deficiency. Molecular analysis of the SRD5A2 gene should be warranted in newborns with abnormal ratio before sex assignment. PMID- 18391526 TI - Combined haploinsufficiency of SF-1 and GATA4 does not reveal a genetic interaction in mouse gonadal development. AB - The nuclear receptor steroidogenic factor 1 (SF-1 or NR5A1) and the zinc finger protein GATA4 mediate key events in the early steps of gonadal development and sex differentiation, presumably by activating the expression of essential target genes. An important SF-1 target in male sex differentiation is the gene encoding the anti-Mullerian hormone (AMH), which induces regression of the Mullerian ducts in the developing male embryo. In cell transfection studies, there is apparent cooperation between GATA4 and SF-1 in the regulation of both human and mouse Amh promoters. We hypothesized that compound haploinsufficiency of both SF-1 and GATA4, by reducing their synergism, might cause a more severe phenotype than that seen in mice that were heterozygous for either SF-1 or Gata4 alone. Surprisingly, in adult and embryonic mice, compound haploinsufficiency of SF-1 and GATA4 caused no gonadal or reproductive abnormalities beyond those seen in SF-1(+/-) mice. Thus, although cooperation between SF-1 and GATA4 very likely is important for regulation of their target genes, such synergy was not revealed in our in vivo studies of gonadal development and function. PMID- 18391527 TI - Synaptonemal complex protein SYCP3 of the rat: evolutionarily conserved domains and the assembly of higher order structures. AB - SYCP3 is a major structural protein component of vertebrate synaptonemal complexes as well as an important determinant of male fertility, at least in mammals. The elucidation of SYCP3 polymerization properties would provide important information towards our understanding as to how synaptonemal complexes are assembled and disassembled during meiotic prophase. To this end we have investigated the possible contribution of different SYCP3 domains to the assembly of higher order structures. We observed that the evolutionarily conserved domains of the molecule (i.e. the alpha-helix together with the two flanking motifs CM1 and CM2) are not only necessary but also sufficient for SYCP3 polymerization. The relevance of these results for reproduction biology is underscored by recent studies showing that the deletion of the very end of the alpha-helix and CM2 leads to meiosis disruption and infertility in humans. PMID- 18391528 TI - Marsupial WT1 has a novel isoform and is expressed in both somatic and germ cells in the developing ovary and testis. AB - The Wilms' tumour 1 gene is essential for the formation of the mouse and human urogenital systems. We characterised this gene and examined its expression throughout gonadal development in a marsupial, the tammar wallaby. WT1 protein was detected in the Sertoli and granulosa cells of the developing testis and ovary, respectively. There was also strong immunostaining in the germ cells of both males and females at all stages of gonadal development. In the adult gonads WT1 appears to be dynamically regulated during spermatogenesis and oogenesis. Tammar WT1 has a novel isoform in which a portion of exon 1 is removed, partially deleting the RNA recognition motif (RRM). Despite its removal, WT1 still localised to RNA rich regions of the oocyte including speckled bodies within the nucleus, in the nucleolus and the perinucleolar compartment. This suggests that the RRM is not required for WT1 co-localisation with RNA. This is also the first report of WT1 in association with the perinucleolar compartment, important for RNA metabolism. Our data suggest that WT1 has a conserved function in both the somatic and germ cell lineages of the gonads of marsupials. PMID- 18391529 TI - Steroid signaling system responds differently to temperature and hormone manipulation in the red-eared slider turtle (Trachemys scripta elegans), a reptile with temperature-dependent sex determination. AB - Many reptiles, including the red-eared slider turtle (Trachemys scripta elegans), exhibit temperature-dependent sex determination (TSD). Temperature determines gonadal sex during the middle of embryogenesis, or the temperature-sensitive period (TSP), when gonadal sex is labile to both temperature and hormones- particularly estrogen. The biological actions of steroid hormones are mediated by their receptors as defined here as the classic transcriptional regulation of target genes. To elucidate estrogen action during sex determination, we examined estrogen receptor alpha (Esr1, hereafter referred to as ERalpha), estrogen receptor beta (Esr2, hereafter referred to as ERbeta), and androgen receptor (Ar, hereafter referred to as AR) expression in slider turtle gonads before, during and after the TSP, as well as following sex reversal via temperature or steroid hormone manipulation. ERalpha and AR levels spike at the female-producing temperature while ovarian sex is determined, but none of the receptors exhibited sexually dimorphic localization within the gonad prior to morphological differentiation. All three receptors respond differentially to sex-reversing treatments. When shifted to female-producing temperatures, embryos maintain ERalpha and AR expression while ERbeta is reduced. When shifted to male-producing temperatures, medullary expression of all three receptors is reduced. Feminization via estradiol (E(2)) treatment at a male-producing temperature profoundly changed the expression patterns for all three receptors. ERalpha and ERbeta redirected to the cortex in E(2)-created ovaries, while AR medullary expression was transiently reduced. Although warmer incubation temperature and estrogen result in the same endpoint (ovarian development), our results indicate different steroid signaling patterns between temperature- and estrogen-induced feminization. PMID- 18391530 TI - Expression of the male determining gene dmrt1bY and its autosomal coorthologue dmrt1a in medaka. AB - Dmrt1bY (aka dmy) has been identified as the master male sex-determining gene of medaka by positional cloning and genetic evidence. How this gene on the molecular level determines the differentiation of the gonad primordium towards a testis, however, is unknown. To better understand the roles of dmrt1bY and its autosomal counterpart dmrt1a in the process of sex determination, a detailed expression analysis by quantitative real-time RT-PCR was performed showing non-organ specific expression of dmrt1bY during embryogenesis and low expression in adult testes. Surprisingly a high expression of dmrt1bY was noted in the spleen of males. Dmrt1a is exclusively expressed in adult testes, 50 times higher than dmrt1bY. Dmrt1bY mRNA and protein have short half-lives and the GFP fusion protein products of both dmrt1 genes are localized to the nucleus. These features are consistent with the suggestion that Dmrt1 proteins might function as DNA binding transcription factors. PMID- 18391531 TI - Mapping of sox2 and sox14 in tilapia (Oreochromis spp.). AB - Sox genes encode transcription factors that are involved in a variety of embryonic developmental pathways. Sox2 and Sox14 are located on the same chromosomal arm in several mammalian and bird species and on the basis of comparative maps were suggested as candidate genes for the major sex-determining locus on tilapia LG3. We have sequenced the sox2 and sox14 genes in four tilapia species and mapped them to different chromosomes, LG17 and LG23 respectively. Although excluded as being one of the major sex-determining genes so far mapped in tilapia, sox14 did fall within a QTL region for growth, stress response, embryonic mortality and a minor effect on sex determination. PMID- 18391532 TI - Ellobius lutescens: sex determination and sex chromosome. AB - The mole vole Ellobius lutescens is an interesting animal, not only concerning its sex determination mechanism without the Y-chromosomal Sry gene, that triggers sex determination in nearly all other mammalian species, but also regarding the karyotype with an odd number of chromosomes, being identical in male and female animals. The odd chromosome represents the X chromosome, and therefore, even males do not have a Y chromosome. We present an overview of a search for candidate genes of male sex determination in the mole vole Ellobius lutescens. A singular X raises questions about the need for X chromosome inactivation in female cells. We present preliminary data that support a hypothesis that the E. lutescens Xist gene may be degenerated and thus non-functional. PMID- 18391533 TI - Predictive value of anatomical findings and karyotype analysis in the diagnosis of patients with disorders of sexual development. AB - We assessed the predictive value of anatomical findings and karyotype for establishing a diagnostic orientation in patients with disorders of sex development (DSD). We performed a retrospective chart analysis of 228 patients, grouped into 4 categories: 46,XX DSD, non-dysgenetic testicular DSD, dysgenetic testicular DSD and ovotesticular DSD. Degree of virilisation, presence of vagina, presence of palpable gonads, size of gonads and a plain karyotype was available for all cases. 46,XX DSD due to congenital adrenal hyperplasia counted for 59.2% of the cases, non-dysgenetic testicular DSD for 13.6%, dysgenetic testicular DSD for 21.5% and ovotesticular DSD for 5.7%. Excluding congenital adrenal hyperplasia (CAH), a karyotype with at least one 46,XX cell line had a high diagnostic efficiency for ovotesticular DSD. In these patients, anatomical findings were not as useful to predict the gonadal phenotype. The existence of a 45,X cell line predicted with very high efficiency dysgenetic testicular DSD. Genital palpation was only partially helpful to predict the existence of testicular tissue. Non-dysgenetic testicular DSD could be ruled out with high efficiency in patients with an abnormal karyotype. Anatomical findings were helpful in 46,XY patients: palpated masses predicted non-dysgenetic testes with high accuracy. In all cases assessment of gonadal volume was less useful. PMID- 18391534 TI - Altered transcription profiles of key-enzymes of androgen biosynthesis in genital skin fibroblasts from patients with 46,XY disorders of sex development (DSD). AB - Normal synthesis and action of androgens is essential for normal male sex differentiation. 17Beta-hydroxysteroid dehydrogenase (17beta-HSD) and 5alpha reductase isoenzymes play essential roles in normal androgen biosynthesis. We hypothesized that differences in expression of these enzymes in genital skin could contribute to the pathogenesis of 46,XY disorders of sex development (DSD). We investigated the mRNA transcription patterns of 17beta-hydroxysteroid dehydrogenase-isoenzymes type 1, 2, 3, 4, 5, 7, and 10, 5alpha-reductase type 1 and 2 and the androgen receptor in genital skin fibroblasts from foreskin and scrotal skin obtained from healthy males and patients with unclassified 46,XY DSD. mRNA expression was semi-quantified by real-time PCR. Although no systematic differences of gene expression of any enzyme between normal controls and hypospadias patients could be detected, we found in nearly half of all investigated patients' samples noticeable differences in the transcription profiles of 17beta-hydroxysteroid dehydrogenase type 5. In scrotal skin samples of patients a significantly higher transcription of the androgen receptor was detected. A role for an altered expression pattern of different enzymes of steroidogenesis in the etiology of genital malformations in some patients may be postulated. PMID- 18391535 TI - WT1-mediated gene regulation in early urogenital ridge development. AB - The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen. As a transcription factor, WT1 fulfills its function by regulating a set of target genes. With respect to gonad development only few in vivo WT1 targets, e.g. steroidogenic factor 1 (SF1) have been identified so far. To get a comprehensive view of WT1 targets in the gonad, we compared gene expression in urogenital ridges of wild-type and Wt1(-/-) embryos. We found almost 150 genes differentially expressed higher than factor three, using microarray analysis. To confirm these results we performed quantitative real-time RT-PCR for many genes and observed a high degree of concordance between microarray and real-time RT-PCR results. Employing in situ hybridization we found 'WT1 activated genes' to be expressed in gonads, mesonephroi and coelomic epithelium--those parts of the urogenital ridge with Wt1 expression. Interestingly, many of the differentially expressed genes are known to show sex-specific expression at later time-points. These results provide a basis for investigation of developmental pathways in the urogenital ridge downstream of WT1 and for identification of new candidate genes involved in early urogenital ridge development. For example we provide a first potential target of WT1 in the coelomic epithelium--Muc16, and a gene regulated by the WT1 target SF1--Gata4. PMID- 18391536 TI - Expression of putative sex-determining genes during the thermosensitive period of gonad development in the snapping turtle, Chelydra serpentina. AB - Modes of sex determination are quite variable in vertebrates. The developmental decision to form a testis or an ovary can be influenced by one gene, several genes, environmental variables, or a combination of these factors. Nevertheless, certain morphogenetic aspects of sex determination appear to be conserved in amniotes. Here we clone fragments of nine candidate sex-determining genes from the snapping turtle Chelydra serpentina, a species with temperature-dependent sex determination (TSD). We then analyze expression of these genes during the thermosensitive period of gonad development. In particular, we compare gene expression profiles in gonads from embryos incubated at a male-producing temperature to those from embryos at a female-producing temperature. Expression of Dmrt1 and Sox9 mRNA increased gradually at the male-producing temperature, but was suppressed at the female-producing temperature. This finding suggests that Dmrt1 and Sox9 play a role in testis development. In contrast, expression of aromatase, androgen receptor (Ar), and Foxl2 mRNA was constant at the male producing temperature, but increased several-fold in embryos at the female producing temperature. Aromatase, Ar, and Foxl2 may therefore play a role in ovary development. In addition, there was a small temperature effect on ER alpha expression with lower mRNA levels found in embryos at the female-producing temperature. Finally, Dax1, Fgf9, and SF-1 were not differentially expressed during the sex-determining period, suggesting these genes are not involved in sex determination in the snapping turtle. Comparison of gene expression profiles among amniotes indicates that Dmrt1 and Sox9 are part of a core testis determining pathway and that Ar, aromatase, ER alpha, and Foxl2 are part of a core ovary-determining pathway. PMID- 18391537 TI - The Mullerian duct: recent insights into its development and regression. AB - Several recent publications have contributed to our understanding of the processes involved in development of the Mullerian ducts in both sexes and regression of these structures in male embryos. Additionally, new insights in the regulation of the anti-Mullerian hormone (AMH) signaling pathway, the pathway, which mediates the male specific degeneration of Mullerian ducts, have been gained. It has become clear that the Mullerian duct is formed by invagination of the coelomic epithelium and elongates primarily by proliferation. Later on cells of the coelomic epithelium perform epithelial to mesenchymal transition and move around the epithelium of the Mullerian duct to induce degeneration of this structure in male embryos. Besides AMH and its specific type II receptor AMHR2 two different type I receptors as well as different SMAD family members have been shown to be involved in the AMH signaling cascade. Other factors including WT1, WNT7a, beta-catenin and MMP2 act upstream and downstream of AMH signaling. Here we try to draw an overall picture of Mullerian duct formation and regression by integrating the recent literature in the field. PMID- 18391538 TI - Sex determination in birds: HINTs from the W sex chromosome? AB - Sex in birds is controlled genetically (ZZ male: ZW female), but the genetic mechanism remains unclear. While some evidence points to the involvement of Z sex chromosome dosage, other data favour a dominant female-determining gene carried on the W sex chromosome. An intriguing candidate gene located on the chicken W chromosome is HINTW, which encodes an aberrant form of a hydrolase enzyme. In chicken embryos, HINTW is strongly expressed in the gonads and other tissues of ZW (female) embryos. In vitro biochemical data show that HINTW can interfere with the action of a Z-linked orthologue, HINTZ, which is a bona fide hydrolase enzyme. HINTW is conserved among carinate (flying) birds, and recent molecular analysis indicates that it has undergone positive selection over evolution. However, a differentiated HINTW gene appears to be absent in the flightless ratites. This review examines the evidence for and against a role for HINTW in avian sex determination. PMID- 18391539 TI - A cellular study of human testis development. AB - This study catalogs the cellular events underlying the formation of a human testis. These events were identified by immunocytochemistry using antibodies that served as markers for specific cell types, then contrasted with the events occurring in the developing mouse testis. The presence of germ cells in the embryonic gonadal ridge and of coelomic epithelial cells that give rise to Sertoli cells was observed at 7 weeks. This was followed by the appearance of Sertoli cells in testicular tubules and of Leydig cells at 9 weeks and by the appearance of vascular endothelial cells and peritubular myoid cells at 12 weeks. Overall the temporal sequence of events in humans was similar, albeit longer, than what occurs in mice. Notably, Leydig cell differentiation occurs earlier in the sequence of events and germ cell maturation occurs during fetal life. The candidate testis-determining genes, FGF9, GATA4, FOG2, EMX2, and CBX2 were expressed at 7 weeks suggesting a role in early gonadal development, such as that observed in mice. In addition, expression of FGF9 in germ cells following testis determination suggests a role in germ cell maturation. PMID- 18391540 TI - Detection of the G34R mutation in the 5 alpha reductase 2 gene by allele specific PCR and its linkage to the 89L allele among Egyptian cases. AB - The 5 alpha-reductase type 2 deficiency is an autosomal recessive disorder of sexual development among 46,XY individuals. In Egypt, there is a prevalence of a G34R disease underlying mutation. This study aimed to devise a rapid diagnostic method based on allele specific PCR (AS-PCR) and a linked polymorphism (V89L). The results showed that one set of primers was capable to differentiate between normal, heterozygous, and affected individuals efficiently. All 34R mutation carrying sequences had 100% linkage to the 89L allele, contrasting normal ones with low 89L frequencies. This linkage infers a founder effect among Egyptians having G34R mutation. PMID- 18391541 TI - Male-specific function of Dmrt7 by sexually dimorphic translation in mouse testis. AB - Dmrt7 is known to be an essential gene for spermatogenesis but not for oogenesis despite mRNA expression in both testis and ovary. In this study, we examined further expression of Dmrt7 transcript and protein. Northern blot and RT-PCR analysis revealed that there was an alternative splicing variant possessing the entire sequence of intron 1 in adult testis (intron 1 variant), in addition to the mature form of mRNA. In fetal ovary, the intron 1 variant was not expressed whereas the fully spliced form of mRNA was expressed. Immunohistochemical analyses demonstrated that DMRT7 protein was present only in spermatocytes of adult testis but not in fetal ovary. In situ hybridization analyses revealed that the fully spliced form of Dmrt7 mRNA as well as the intron 1 variant were expressed in spermatogonia, spermatids and Sertoli cells in addition to spermatocytes. We also found that poly(A) tails of Dmrt7 mRNA underwent modification of its length from 70 to 440 bp long. Unlike Arbp mRNA, the size variation of poly(A) tails was observed in immature testis in which spermatids were absent. In this study, we demonstrated that Dmrt7 had unique sexually dimorphic expression patterns in transcripts that associated with spermatocyte specific translation, but not in ovary. PMID- 18391542 TI - Comparative analysis of anti-mouse SRY antibodies. AB - The Y chromosome gene SRY is the initiator of male sexual differentiation in mammals, but the molecular and cellular mechanisms operating downstream of SRY remain undefined. A deeper understanding of these issues relies on the ability to visualize SRY protein endogenously under a number of experimental conditions. Here we compare the specificity and effectiveness of several available antibodies to mouse SRY. Two antibodies cross-reacted with other SOX proteins in immunofluorescence analyses of transfected cells, and one of these two was unable to detect SRY on Western blots. A third antibody was both avid and specific, and was able to detect endogenous SRY in developing Sertoli cells in mouse genital ridges. Our findings underline the need to distinguish between useful and spurious reagents for biochemical and immunolocalization studies involving mouse SRY protein. PMID- 18391543 TI - Fugu (Takifugu rubripes) sexual differentiation: CYP19 regulation and aromatase inhibitor induced testicular development. AB - In order to assess the involvement of aromatase CYP19 isoforms and endogenous sex steroids in gonadal sex differentiation and development of the Japanese fugu (Takifugu rubripes), an aromatase inhibitor (AI, fadrozole) was administered to developing fishes from the 'first feeding' till the 100th day after hatching. It was observed that ovarian cavity formation was inhibited by fadrozole at doses of 500 and 1000 microg/g diet, which was followed by testicular differentiation in all treated fugu. In the non-treated fugu, CYP19A was predominantly expressed in the ovary and CYP19B in the brain (in both sexes), although both were expressed interchangeably at low levels. An exceptionally high expression of CYP19B was also evident in testis throughout the study period. Both forms of CYP19 mRNA showed low levels of expression in brain and gonad with no significant differences between the two AI treatments. AI treatment inhibited CYP19A mRNA in trunk during the crucial period of ovarian cavity formation and CYP19B in gonad and brain by the end of gonadal sex differentiation. An elevation of testosterone and 11-ketotestosterone was observed which can be associated with the down regulation of the circulating 17beta-estradiol production during the AI treatment period. After stopping AI treatment, both circulating estrogen and androgen were normalized. The current results suggest that suppression of CYP19A before and during morphological sex differentiation inhibits ovarian cavity formation in fugu. Furthermore, non-detectable limits of 17beta-estradiol and high testosterone levels by the end of the gonadal differentiation period can be ascribed to inhibition of CYP19B, suggesting that conversion of 17beta-estradiol from testosterone is plausibly regulated by CYP19B, and that this factor (CYP19B) may play an important role in AI-induced testicular development after gonadal sex differentiation through regulation of the testosterone-17beta-estradiol balance in fugu. PMID- 18391544 TI - An evolutionary witness: the frog rana rugosa underwent change of heterogametic sex from XY male to ZW female. AB - There are two basic types of heterogamety for genetic sex determination in animals and plants: male heterogamety (XX/XY) and female heterogamety (ZZ/ZW). Although apparently in opposition, the two distinct types may in fact be interchangeable. For example, in amphibians it has been shown that the heterogametic sex was originally female and may have become male at some branching point in their phylogenetic evolution. In particular, there is evidence that the male heterogametic sex determination of the frog Rana rugosa returned to its previous female state during speciation that occurred when the distribution range of the frog expanded across Japan. This change is quite recent in the phylogenetic time scale. This paper presents a review of the sex chromosomes and sex determination in the frog R. rugosa, an evolutionary witness proving the viability of changing heterogametic sex, and introduces recent findings and on going studies in the frog. Change of the heterogametic sex will also be discussed, relating data from frogs (Rana) and other animals to the replacement of a master sex-determining gene in the course of speciation. PMID- 18391545 TI - Sex chromosomes and sex determination in Lepidoptera. AB - The speciose insect order Lepidoptera (moths and butterflies) and their closest relatives, Trichoptera (caddis flies), share a female-heterogametic sex chromosome system. Originally a Z/ZZ (female/male) system, it evolved by chromosome rearrangement to a WZ/ZZ (female/male) system in the most species-rich branch of Lepidoptera, a monophyletic group consisting of Ditrysia and Tischeriina, which together comprise more than 98% of all species. Further sporadic rearrangements created multi-sex chromosome systems; sporadic losses of the W changed the system formally back to Z/ZZ in some species. Primary sex determination depends on a Z-counting mechanism in Z/ZZ species, but on a female determining gene, Fem, in the W chromosome of the silkworm. The molecular mechanism is unknown in both cases. The silkworm shares the last step, dsx, of the hierarchical sex-determining pathway with Drosophila and other insects investigated, but probably not the intermediate steps between the primary signal and dsx. The W chromosome is heterochromatic in most species. It contains few genes and is flooded with interspersed repetitive elements. In interphase nuclei of females it is readily discernible as a heterochromatic body which grows with increasing degree of polyploidy in somatic cells. It is used as a marker for the genetic sex in studies of intersexes and Wolbachia infections. The sex chromosome system is being exploited in economically important species. Special strains have been devised for mass rearing of male-only broods in the silkworm for higher silk production and in pest species for the release of sterile males in pest management programs. PMID- 18391546 TI - Chromosome abnormalities and Yq microdeletions in infertile italian couples referred for assisted reproductive technique. AB - This study analyses the prevalence of karyotype aberrations and Yq microdeletions in infertile couples undergoing intracytoplasmic sperm injection (ICSI). Before undergoing ICSI, each partner of 470 infertile couples was screened for karyotype aberrations by QFQ-banding technique on peripheral blood lymphocytes; male partners were also screened for Yq microdeletions. In 2.55% of the couples karyotype aberrations were found including numerical and structural alterations of autosomes and sex chromosomes. The female group had a high prevalence of low level sex chromosome mosaicism (1.28%) and 5 cases of structural autosomal abnormalities (1.06%). The male group had 7 structural abnormalities of the autosomes (1.49%), 2 supernumerary marker chromosomes (0.42%), one case of low level gonosomal mosaicism (0.21%), and 2 cases of Y chromosome inversion (0.42%). Eight cases of Yq microdeletions (1.70%) were also found. Screening for genetic factors, chromosomal abnormalities and Yq microdeletions is indicated for couples undergoing assisted reproductive techniques due to the higher prevalence of these factors in infertile couples compared to the population as a whole although different chromosome aberrations have been reported elsewhere. PMID- 18391547 TI - Small supernumerary marker chromosomes (sSMC) in patients with a 45,X/46,X,+mar karyotype - 17 new cases and a review of the literature. AB - Small supernumerary marker chromosomes (sSMC) can appear in a numerically normal 'basic karyotype', but also in a numerically abnormal one like a Turner syndrome karyotype (= sSMC(T)). Here we present 17 new cases with such a mos 45,X/46,X,+mar karyotype. Moreover we reviewed all 512 cytogenetically similar cases available from the literature and supply for the first time data on occurrence, shapes and subgroups of this rare cytogenetic entity. sSMC(T) are very rare in the common population (1:100,000) - however, they can be observed with a 45- and even 60-times higher frequency in infertile and (develop)mentally retarded patients, respectively. Even though sSMC(T) derive from one of the gonosomes in >99% of the cases, there are also exceptional reports on sSMC(T) derived from one of the autosomes. The majority of sSMC(T)(X) form ring chromosomes, while most sSMC(T)(Y) are inverted duplicated/isodicentric chromosomes. Although >500 sSMC(T) are reported, a detailed characterization of the chromosomal breakpoints is only given for a minority. Thus, more cases with detailed (molecular) cytogenetic marker chromosome characterization are needed to provide information on formation and effects of an sSMC(T). PMID- 18391548 TI - The reproductive tissue specific cystatin subgroup of genes: expression during gonadal development in wildtype and testatin knockout animals. AB - Testatin has been implicated in fetal testis development due to its restricted expression in pre-Sertoli cells immediately after the onset of Sry gene expression. However, testatin knockout mice showed normal testis development and fertility. We investigated the spatial and temporal expression pattern of the Cres/testatin subgroup of genes, including the novel gene Cstl1/Cres4, in fetal mouse gonads and in adult testis, epididymis and ovary. The genes are related to the family 2 cystatins of protease inhibitors. Using real-time PCR and in situ hybridization we could show that 4 subgroup genes, testatin, CstSC, CstTE-1/Cres3 and Cres are expressed in fetal testis. We also confirmed the expression of testatin, CstE2, CstSC, CstTE-1/Cres3, Cres, CstT and Cstl1/Cres4 in adult testis and CstE2, CstTE-1/Cres3, Cres and CstE1/Cres2 in adult epididymis. In testatin knockout animals, the expression of CstE2 was heavily downregulated in adult testis, but not in adult epididymis, compared to wildtype controls. In conclusion, an explanation for the lack of phenotype in testatin knockout mice could be functional redundancy with another member of the Cres/testatin subgroup. The most likely candidate/s would be CstSC, CstTE-1/Cres3 or Cres as they are expressed in the fetal testicular tubules in early testis differentiation together with testatin. PMID- 18391549 TI - Detection of human papillomavirus DNA by DNA chip in breast carcinomas of Korean women. AB - It remains unclear whether there is an association between human papillomavirus (HPV) infection and human breast cancer. The aim of this study was to investigate the presence of HPV DNA in breast carcinomas of Korean women and to examine the possible association between HPV and breast cancer development. For this purpose, HPV DNAs from 154 patients, including 123 patients with breast carcinoma and 31 with intraductal papilloma, and nipple tissue from 27 cancer patients were examined using the DNA chip method. HPV DNA was detected in 8 breast carcinomas (6.5%) but in no intraductal papilloma. All detected HPV genotypes were of high risk groups. There was a slightly increased incidence in papillary carcinomas (11.5%) and invasive ductal carcinomas with adjacent intraductal papillomas (11.8%) compared to the other histological subtypes (3.2-4.3%), although the difference was not statistically significant (p = 0.126). The presence of HPV DNA was not correlated with specific prognostic predictors of disease. High-risk HPV DNA sequences were present in 6.5% of Korean patients with breast tumors. However, this study could not demonstrate whether or not such HPVs directly contribute to the development of breast cancer. PMID- 18391550 TI - Activation-induced cytidine deaminase expression in gastric cancer. AB - Helicobacter pylori increases the risk of gastric cancer development and triggers aberrant expression of activation-induced cytidine deaminase (AID). The goal of the present study was to investigate whether AID expression is involved in the development or progression of gastric cancer and the nuclear expression of p53 protein in cancer cells. We examined the expression pattern of the AID and p53 proteins in 186 gastric adenocarcinomas by immunohistochemistry. In order to investigate the molecular mechanism of AID expression, we also searched for mutations in the AID gene by single-strand conformational polymorphism and sequencing methods. In 186 sporadic gastric cancers, AID expression was detected in the 73 corresponding normal gastric mucosa and in 50 gastric cancers. Statistically, the expression of AID protein was not associated with clinicopathological parameters, including tumor size, location, differentiation and lymph node metastasis (p > 0.05). Interestingly, a significant association was observed between AID and the nuclear expression of p53 (p = 0.0094). Mutational analysis revealed no mutation in the AID gene in the gastric cancers. These results suggest that aberrant expression of the AID protein may contribute to the development of gastric cancers and induce p53 nuclear expression. PMID- 18391551 TI - The prognostic impact of blood group-related antigen Lewis Y and the ABH blood groups in resected non-small cell lung cancer. AB - The blood group antigen Lewis Y is expressed on epithelial tumors of the respiratory, digestive and reproductive system. Despite being regarded as an attractive target for immunotherapy, its function is still not well defined and its prognostic value remains a subject of discussion. Eighty-three paraffin embedded tissue sections of non-small cell lung cancer (NSCLC) patients in stage I-IIIa, who underwent surgical resection of the primary tumor (73% male; 43% adenocarcinoma), were stained with a new, highly specific monoclonal antibody against Lewis Y (clone A70-C/C8). A positive Lewis Y expression was observed in 51% of patients; adenocarcinomas were favorably stained (67%). Multivariate analysis identified stage I, blood group A or AB and Lewis Y expression on tumor cells to be independent markers for improved survival after tumor resection (p = 0.024, 0.043, 0.003, respectively). In summary, unlike in several previous studies the presence of Lewis Y on tumor cells is a favorable prognostic factor in this cohort of resected NSCLC patients. Coexisting blood group antigen A may be of additional positive prognostic impact. We hypothesize that related blood group antigens both on tumor cells and in peripheral blood may have an underestimated function for progression in resected NSCLC. PMID- 18391552 TI - A simple immunomonitoring procedure for mRNA-loaded dendritic cell therapy. AB - To develop a simple immunomonitoring method for dendritic cell therapy using messenger RNA (mRNA) as antigen, we evaluated whether Mycobacterium tuberculosis antigen 85A (Ag85A) mRNA-transfected peripheral blood mononuclear cells (PBMCs) could be used to stimulate the induction of interferon (IFN)-gamma-producing T cells. PBMCs from 10 healthy donors were cocultured with autologous PBMCs transfected with mRNA overnight, and the number of IFN-gamma-producing T cells was measured by flow cytometry. IFN-gamma-producing CD4+ and CD8+ T cells were detected in 4 and 5 donors, respectively. PBMCs from 3 donors with negative results were then cocultured with Ag85A mRNA-transfected autologous PBMCs for 1 week to achieve in vitro primary induction of Ag85A-specific T cells. After restimulation with freshly prepared stimulator cells, a small but significant number of IFN-gamma-producing CD8+ T cells was detected. The induction of IFN gamma-producing CD8+ T cells by overnight coculture was completely abolished by anti-class II or anti-interleukin-12 antibodies, whereas it was partially inhibited by anti-class I antibody. These data suggest that Ag85A mRNA transfected PBMCs induce specific IFN-gamma-producing T cells and might be applicable for immunomonitoring of mRNA-loaded dendritic cell therapy. PMID- 18391553 TI - Introduction: new developments in the area of sexual dysfunction(s). AB - New developments in the area of sexual dysfunction, e.g. epidemiology and pharmacological treatment, are reviewed. Areas where new developments/changes are needed, such as diagnosis of sexual dysfunction and research methodology, are also briefly discussed. PMID- 18391554 TI - Clinical and research evaluations of sexual dysfunctions. AB - Evaluations of the nature of an individual's sexual functioning are done with relative frequency in today's clinical healthcare environment, often prompted by sequelae of a medical disorder (e.g. diabetes, breast cancer), surgical procedure (e.g. bilateral oophorectomy), or prescribed medications (SSRI antidepressant, antihypertensive). In addition, an increasingly large number of evaluations of sexual functioning are done in association with clinical trials research in the rapidly emerging field of sexual medicine. Although clinical and research assessments of sexual functioning share many similarities, they can also be quite different, with distinct perspectives and goals regarding the assessment process. Research evaluations tend to be narrowly focused on an index condition, with highly defined patient characteristics, and long lists of inclusion/exclusion criteria. Treatment regimens are defined by research protocol a priori, and assigned through randomization, with few options for alternatives. The process results in a rigorously defined sample of patients, but one that may be hard to generalize from to real world clinical reality. Clinical sexual evaluations, on the other hand, tend to arise in a much broader health context. There are no exclusionary patient characteristics; medical, psychological, and relationship complexities all form the context in which the patient's sexual functioning occurs, and must be factored in to the clinical evaluation and treatment recommendations. Regardless of which context gives rise to the evaluation, there are three basic sources of data that we draw from: psychometric assessment, clinical interview and laboratory assays. The clinician doing a sexual evaluation must assimilate and integrate the data from these three sources in an iterative process and arrive at a cogent diagnosis and treatment plan. PMID- 18391555 TI - Management of hypoactive sexual desire disorder. AB - Epidemiological studies have found that problems with sexual desire are one of the most common sexual complaints in females. Such complaints are far less common in males. In women, problems with sexual desire have been found to be associated with age, relationship duration, relationship distress, and complaints of anxiety and depression. Evidence-based interventions include cognitive behavioral therapy and androgen therapy. Endocrinopathies are common causes of male problems with libido. There is minimal evidence available concerning the treatment of psychological etiologies of low sexual desire in men. PMID- 18391556 TI - Erectile dysfunction. AB - Erectile dysfunction is a common problem affecting sexual function in men. Approximately one in 10 men over the age of 40 is affected by this condition and the incidence is age related. Erectile dysfunction is a sentinel marker for several reversible conditions including peripheral and coronary vascular disease, hypertension and diabetes mellitus. Endothelial dysfunction is a common factor between the disease states. Concurrent conditions such as depression, late-onset hypogonadism, Peyronie's disease and lower urinary tract symptoms may significantly worsen erectile function, other sexual and relationship issues and penis dysmorphophobia. A focused physical examination and baseline laboratory investigations are mandatory. Management consists of initiating modifiable lifestyle changes, psychological and psychosexual/couples interventions and pharmacological and other interventions. In combination and with treatment of concurrent comorbid states, these interventions will often bring about successful resolution of symptoms and avoid the need for surgical interventions. PMID- 18391557 TI - Recent advances in the classification, neurobiology and treatment of premature ejaculation. AB - Premature ejaculation (PE) is distinguished as a 'complaint' versus a 'syndrome'. Complaints of PE in men with normal ejaculation time durations are highly prevalent among the general male population. However, PE syndromes characterized by a rigid pattern of very short ejaculation times have a much lower prevalence. A major insufficiency of the DSM-IV-TR definition of PE is the absence of a cut off point of its short ejaculation time criterion. Recently, a new classification of PE has been proposed for the pending DSM-V. It has been proposed to distinguish four PE categories; lifelong PE, acquired PE, natural variable PE and premature-like ejaculatory dysfunction. The syndromes differ in ejaculation time duration, frequency and pattern of PE complaints, and its course in life. The difference in underlying pathophysiology and etiology of the different syndromes determines the first choice of treatment. Lifelong PE need to be treated by medication, acquired PE by treatment of its medical cause, medication or psychotherapy. Psychoeducation and reassurance is indicated for men with natural variable PE. Finally, psychotherapy and/or counselling is the first choice of treatment of men with premature-like ejaculatory dysfunction. PMID- 18391558 TI - The impact of physical illness on sexual dysfunction. AB - Sexuality is the ultimate union of mind and body. Sexual dysfunction is often the first manifestation of physical illness but is often not inquired about on routine review of symptoms. This is, in large part, due to the health care providers' lack of knowledge in diagnosis and treatment of sexual impairment as well as their discomfort with this sensitive topic. However, sexual well-being is an important determinant of quality of life and many medically ill patients find sexual intimacy to be an essential mode of communication with their partners. This chapter attempts to methodically delineate physical illnesses causing sexual dysfunction by organ system. Neurologic, endocrinologic, cardiovascular and pelvic illnesses are discussed as to their impact on sexual health. Diagnostic and established treatment strategies are also reviewed. Breast cancer, rheumatoid arthritis and psoriasis are touched upon. Although not a disease, pregnancy and its unique impact on sexuality is also discussed. Not only the disease itself but the treatment prescribed may also cause sexual impairment. Thus, a separate section on medications that impair sexual functioning is presented. A table of common medications as a quick reference to their effects on each stage of the sexual cycle is also provided. PMID- 18391559 TI - The impact of mental illness on sexual dysfunction. AB - Sexual dysfunction is prevalent among psychiatric patients and may be related to both the psychopathology and the pharmacotherapy. The negative symptoms of schizophrenia limit the capability for interpersonal and sexual relationships. The first-generation antipsychotics cause further deterioration in erectile and orgasmic function. Due to their weak antagonistic activity at D2 receptors, second-generation antipsychotics are associated with fewer sexual side effects, and thus may provide an option for schizophrenia patients with sexual dysfunction. Depression and anxiety are a cause for sexual dysfunction that may be aggravated by antidepressants, especially selective serotonin reuptake inhibitors (SSRIs). SSRI-induced sexual dysfunction may be overcome by lowering doses, switching to an antidepressant with low propensity to cause sexual dysfunction (bupropion, mirtazapine, nefazodone, reboxetine), addition of 5HT2 antagonists (mirtazapine, mianserin) or coadministration of 5-phosphodiesterase inhibitors. Eating disorders and personality disorders, mainly borderline personality disorder, are also associated with sexual dysfunction. Sexual dysfunction in these cases stems from impaired interpersonal relationships and may respond to adequate psychosexual therapy. It is mandatory to identify the specific sexual dysfunction and to treat the patients according to his/her individual psychopathology, current pharmacotherapy and interpersonal relationships. PMID- 18391560 TI - Psychotropics and sexual dysfunction: the evidence and treatments. AB - Sexual dysfunction is common in the general population and even more common in the mentally or medically ill. Because mentally ill patients often receive psychotropics, many of which affect sexual functioning, a patient's pre-existing sexual difficulties are often compounded, and these adverse effects may contribute to psychological difficulties or medication discontinuation. The effects of antidepressants, antipsychotics, mood stabilizers and anxiolytics on sexual functioning are critically reviewed. When possible, the types of sexual dysfunction (e.g. desire, arousal, or orgasm) induced by the drug is described. Treatments for drug-induced sexual function are described, but few controlled studies show benefit. Only sildenafil stands as a convincing treatment for drug induced sexual dysfunction. The paper focuses on the placebo-controlled clinical trials that specifically evaluated sexual functioning in patients treated with psychotropics. Controlled studies are few for all the agents, though best for the antidepressants and antipsychotics. The exact magnitude and phase of sexual functioning affected remains to be elucidated for most psychotropic drugs. Although all phases of sexual functioning may be impaired by psychotropics, selective serotonin reuptake inhibitor antidepressants appear to primarily affect orgasm, while antipsychotics primarily affect desire. There is insufficient evidence to make conclusions about the anxiolytics or mood stabilizers. PMID- 18391561 TI - Drugs of abuse and sexual functioning. AB - The use of mind-altering substances can be found in very different cultures and traced back thousands of years; the same is true for the searching of drugs that could increase sexual functioning. In this text, we explore the relation between drugs of abuse and sexuality in three domains: drugs and sexual dysfunctions, drugs and risky sexual behavior and drugs used as sexual aids. Although some drugs can increase sexual response in the early stages of the addiction career, particularly in those with a previous sexual dysfunction, the chronic use of substances tends to deteriorate all stages of sexual response in both male and female abusers. There is sufficient evidence for considering that drug use before or during sexual intercourse can, in certain circumstances, elevate the risk of unwanted pregnancy or sexually transmitted diseases. Specific prevention strategies should be addressed to this population. Some psychotropic drugs are sometimes used as sexual aids. This can have some risks and should alert the therapist to a possible underlying and undiagnosed sexual problem. PMID- 18391562 TI - Contribution of imaging to our understanding of sexual function and dysfunction. AB - The physiology of sexual function and dysfunction is complex and involves biologic, psychological, emotional and social factors in both men and women. As a result, multiple investigative tools are needed to achieve a thorough and comprehensive understanding of these processes. Recently, there have been many advances in the field of medical imaging; many of these imaging techniques have been applied to the study of sexual function and have allowed us to obtain an additional perspective of sexual processes in the research setting. The purpose of this chapter is to review and discuss these techniques with an emphasis on how they contribute to our understanding of sexual function and dysfunction. PMID- 18391563 TI - Philadelphia-positive chronic-phase chronic myeloid leukaemia after long-term remission of Philadelphia-negative acute lymphoblastic leukaemia. PMID- 18391564 TI - Testing the diagnostic value of electrical ear canal stimulation in cochlear implant candidates by functional magnetic resonance imaging. AB - Prior to cochlear implant (CI) surgery in children, the integrity of the auditory pathway is sometimes assessed by electrical ear canal stimulation (ECS). However, the evaluation of reactions as auditory is subjective. To test the prognostic value of ECS, functional magnetic resonance imaging (fMRI) was performed during ECS vicariously in 18 adult CI candidates. Activation of the primary auditory cortex was detected in 9 of 16 cases when auditory sensations during ECS occurred, and tended to be more bilaterally distributed in CI candidates than in normal-hearing controls. ECS sensations only tended to correlate with fMRI activations. However, solely frequency discrimination during electrical stimulation predicted CI outcome, but neither other auditory sensations nor fMRI activations did so satisfactorily, which limits the diagnostic value of these measures. Instead, preoperative residual hearing (nonamplified and amplified) was a robust predictor for CI benefit. PMID- 18391565 TI - Ultrastructural changes in otoconia of osteoporotic rats. AB - The etiology of benign paroxysmal positional vertigo (BPPV) remains obscure in many cases and women are affected more often than men. A recent prospective study, performed in women >50 years of age suffering from recurrent BPPV, showed associated osteopenia or osteoporosis in a large percentage of these patients. These results suggested the possible relationship between recurrent BPPV and a decreased fixation of calcium in bone in women >50 years. To test this hypothesis, an experimental study was performed in adult female rats. Utricular otoconia of female rats in which osteopenia/osteoporosis was induced by bilateral ovariectomy (OVX) were compared to those of sham-operated adult females rats (SHAM), as control group. FIRST STUDY: The morphology of theutricles of OVX and SHAM rats was analyzed with scanning electron microscopy. In osteopenic/osteoporotic rats, the density of otoconia (i.e. the number of otoconia per unit area) was decreased (p = 0.036)and their size was increased (p = 0.036) compared to the control group. SECOND STUDY: To test the role of calcium turnover in such morphological changes, utricular otoconia of 2 other groups of OVX and SHAM rats, previously injected with calcein subcutaneously, were examined by conventional and epifluorescence microscopy. In epifluorescence microscopy, labeling with calcein showed no significant fluorescence in either group. This finding was interpreted as a lack of external calcium turnover into otoconia of adult female rats. The ultrastructural modifications of otoconia in osteopenic/osteoporotic female adult rats as well as the role of estrogenic receptors in the inner ear are discussed. The possible pathophysiological mechanisms which support the relationship between recurrent BPPV in women and the disturbance of the calcium metabolism of osteopenia/osteoporosis are debated. PMID- 18391566 TI - Intellectual ability of Mandarin-speaking children using cochlear implants. AB - The aim of this study was to assess the verbal and performance intelligence of young Mandarin-speaking cochlear implant users with the Wechsler Intelligence Scale for Children (WISC). We also analyzed whether related factors helped develop verbal and performance IQs. We studied 60 children implanted with Nucleus 24 devices; the children represented a consecutive sample of every implantee aged 6 or older from 2002 to 2006. All subjects had 1 year to 8 years and 7 months of implant experience. Five children with known neurological, developmental delay and multiple handicaps were excluded. Intellectual functions were evaluated using the Mandarin version of the WISC-III, which includes 5 subsets for verbal IQ (information, comprehension, similarities, arithmetic and vocabulary) and 5 subsets for performance IQ (picture completion, picture arrangement, block design, object assembly and coding). After conversion, we found that the mean verbal IQ was 85.1 +/- 19.9 (range 56-133), and the mean performance IQ was 99.2 +/- 15.9 (range 61-131). The distribution of verbal IQ was significantly different from that of the hearing population. A much larger proportion (32%) falls into the category of 'intellectual deficiency' and a relatively smaller proportion in the 'average' and 'above average' level. As for performance IQ, the distribution was not significantly different from the norms. Regarding the possibly related factors, univariate and multivariate linear regression showed that the verbal IQ was significantly affected by gender (female > male, p = 0.004), side of implantation (bilateral > left > right, p = 0.017) and two speech test scores (PB score, p = 0.036; sentence, p = 0.002), but not by age at implantation (p = 0.621) or length of implant usage (p = 0.480). Only a moderate correlation (r = 0. 49) was found between verbal and performance IQ. In conclusion, we have demonstrated that the performance IQ for the cochlear implant users is not different from their counterparts with normal hearing. However, the distribution of verbal IQ significantly shifts to the left. Our findings suggest that the verbal IQ test may just be another measurement of spoken language outcomes or learning skills and therefore may not represent the 'true intelligence' of these cochlear implant users. PMID- 18391567 TI - Sensitivity to interaural level difference and loudness growth with bilateral bimodal stimulation. AB - The interaural level difference (ILD) is an important cue for the localization of sound sources. The sensitivity to ILD was measured in 10 users of a cochlear implant (CI) in one ear and a hearing aid (HA) in the other severely impaired ear. For simultaneous presentation of a pulse train on the CI side and a sinusoid on the HA side the just noticeable difference (JND) in ILD and loudness growth functions were measured. The mean JND for pitch-matched electric and acoustic stimulation was 1.7 dB. A linear fit of the loudness growth functions on a decibel-versus-microampere scale shows that the slope depends on the subject's dynamic ranges. PMID- 18391568 TI - Molecular characterization of putative modulatory factors in two Spanish families with A1555G deafness. AB - The aim of this work is to characterize possible modifying factors in 2 large families carrying the A1555G mitochondrial mutation. The heteroplasmy of the mutation, the presence of aminoglycosides, the cosegregation with other mitochondrial mutations, the proposed linkage in chromosome 8 and the association with TRMU and MTO1 genes were studied. None of the mentioned modifying factors were related with the phenotype presentation of A1555G mutation. However, TRMU G28T single nucleotide polymorphism is present in 1 of the studied families. PMID- 18391569 TI - Metabonomics study of intestinal transplantation using ultrahigh-performance liquid chromatography time-of-flight mass spectrometry. AB - BACKGROUND: Intestinal transplantation is a potential treatment for patients with irreversible intestinal failure. However, the long-term outcome is still unsatisfactory compared with transplantation of other solid organs because of transplantation rejection. Therefore, early detection and accurate diagnosis of rejection is essential. Recently, metabonomics has become a platform for monitoring of the process of organ transplantation. METHODS: The metabolite profiling in the plasma was measured by ultrahigh-performance liquid chromatography time-of-flight mass spectrometry in a patient with intestinal transplantation and it was analyzed by principal component analysis and SIMCA-P. RESULTS: The differences between the patient and controls were achieved in the principal component analysis score plot. The potential biomarkers of rejection, including lysophosphatidyl choline, phenylalanine and tryptophan, were identified in the transplantation patient. CONCLUSION: The analysis here may provide a fundamental and powerful approach to inspect the disease metabolic status and screening tool for marker candidates. PMID- 18391570 TI - Interleukin-18 in urine and serum of children with idiopathic nephrotic syndrome. AB - BACKGROUND/AIMS: Interleukin (IL)-18, a member of the IL-1 cytokine superfamily, is recognized as an important regulator of immune responses. The aim of our study was to investigate the IL-18 levels in serum and urine from children with idiopathic nephrotic syndrome (INS) during relapse and remission, and to evaluate the role of IL-18 in this disease. METHODS: 67 children with INS, aged 3-16 years, and 15 normal controls were included in the study. The patients were divided into two groups according to activity of the disease: I (n = 37) - INS in relapse, II (n = 30) - INS in remission. Serum and urinary IL-18 were determined by ELISA and in urine related to the urinary creatinine (Cr) concentration. Serum creatinine, protein, albumin and 24-hour proteinuria were measured in children with INS. RESULTS: Urinary IL-18 concentration was significantly higher in group I (213.51 +/- 162.15 pg/mg Cr) compared to group II (64.74 +/- 10.95 pg/mg Cr) and to normal controls (37.03 +/- 4.1 pg/mg Cr, p < 0.001). Serum IL-18 concentration was significantly higher in group I than in the controls (146.4 +/- 30.2 and 113 +/- 10 pg/ml, respectively; p < 0.05); the differences between either groups I and II or group II and controls were not significant. Urinary IL 18 correlated positively with serum IL-18 and with urinary protein excretion, but no correlations were found with other laboratory data. CONCLUSION: Increased serum and urine IL-18 levels were observed during relapse of INS. These findings indicate the association between the active phase of INS and the levels of IL-18 and can suggest the role of this cytokine in the INS development. The changes in urinary IL-18 excretion in the course of INS are connected with the disease activity. PMID- 18391571 TI - Lack of iNOS impairs endothelial function in endothelin-1 transgenic mice. AB - BACKGROUND: Endothelin-1 (ET-1) is one of the most potent biologic vasoconstrictors. Nevertheless, transgenic mice overexpressing ET-1 exhibit normal blood pressure. We hypothesized that in states of ET-1 overproduction, the lack of counterregulatory mediators such as nitric oxide (NO), produced by the inducible NO synthase (iNOS), may critically impair endothelial function and may result in blood pressure elevation. METHODS: We generated crossbred animals of ET transgenic mice (ET+/+) and iNOS knockout mice (iNOS-/-) and evaluated blood pressure and endothelial function in these animals. Endothelium-dependent and independent vascular function was assessed as relaxation/contraction of isolated preconstricted aortic rings to acetylcholine, sodium nitroprusside, and ET-1, alone or in the presence of BQ123 or BQ788. RESULTS: Systolic blood pressure was similar in ET+/+, iNOS-/- and wild-type mice, but was significantly elevated in ET+/+ iNOS-/- crossbred animals versus ET+/+ mice. Maximum endothelium-dependent relaxation was enhanced in ET+/+ mice (95 +/- 5 vs. 78 +/- 5% of preconstriction in wild-type littermates; p < 0.05). Additional knockout of iNOS led to a significant decrease of endothelium-dependent relaxation in combined ET+/+ iNOS-/ animals (75 +/- 6%; p < 0.05 vs. ET+/+ mice). Endothelium-independent relaxation was comparable among all groups. Maximum vascular contraction to ET-1 was reduced in ET+/+ mice (33 +/- 4%), iNOS-/- mice (38 +/- 5%) and ET+/+ iNOS-/- mice (44 +/ 4%) to a similar extent as compared with wild-type littermates (66 +/- 4%; p < 0.05). CONCLUSIONS: Our data show for the first time that in transgenic mice overexpressing human ET-1, additional knockout of iNOS results in impaired endothelium-dependent vasodilatation thus contributing to elevated blood pressure in ET+/+ iNOS-/- animals. PMID- 18391572 TI - New data on the intraindividual variation of cystatin C. AB - BACKGROUND: Cystatin C is a new interesting marker of glomerular filtration rate (GFR). However, data regarding its biological variance are scarce and conflicting. The ability of cystatin C to longitudinally follow renal function in patients therefore remains questionable. METHODS: 12 healthy subjects (6 men and 6 women) were included in the final statistical analysis. Serum creatinine, plasma cystatin C and GFR were measured twice after a 1-week interval on the same day, at the same time, and under the same preanalytical and analytical conditions. GFR was measured with an iohexol method. Serum creatinine was measured with a compensated Jaffe and an enzymatic method. Plasma cystatin C was measured by a particle-enhanced immunonephelometric method. Analytical (CV(A)) and within-subject (CV(I)) variances were classically calculated. RESULTS: CV(A) for creatinine (Jaffe and enzymatic methods) and cystatin C was 2.5, 0.97 and 1.29%, respectively. CV(I) was 5.8, 5 and 4.5% for the Jaffe creatinine, enzymatic creatinine and cystatin C determinations, respectively. CONCLUSION: Our study confirms that intraindividual variation of cystatin C and creatinine are similar. Therefore, from a biological point of view, cystatin C seems as accurate as creatinine for the longitudinal follow-up of renal function in daily clinical practice. PMID- 18391573 TI - Comparison of the long-term efficacy of subcutaneous and sublingual immunotherapies in perennial rhinitis. AB - BACKGROUND: Both sublingual and subcutaneous immunotherapies have a documented clinical efficacy, but only a few comparative studies have been performed. OBJECTIVES: To evaluate and compare the long-term efficacies of subcutaneous and sublingual immunotherapy. METHODS: One hundred and ninety-three patients with house dust mite allergies, out of an original total of 230, were treated with subcutaneous and sublingual house dust mite-specific immunotherapies for 3 years and also observed for 3 years after discontinuation of the treatment. The patients were randomized into 2 groups: the sublingual group (97 patients) and the subcutaneous group (96 patients). The therapy's success was evaluated using the symptom score, skin prick test results, and the nasal allergen challenge score. The patients were evaluated at the beginning of the study, at the end of years 1, 2, and 3, and also at the end of the 1st and 3rd years after discontinuation of the specific immunotherapy treatment. RESULTS: Immunotherapy induced a significant improvement during the treatment and the follow-up period. We found a greater improvement in the subcutaneous group compared to the sublingual group when we looked at the comparative results of the total 6 years. CONCLUSION: We suggest subcutaneous immunotherapy for patients with perennial allergic rhinitis due to the better results that were obtained during our study period. Nevertheless, sublingual immunotherapy is now accepted by WHO as a valid alternative to the subcutaneous route and should be used in all patients who require immunotherapy and do not accept the subcutaneous route of allergen administration. PMID- 18391574 TI - Questionnaire analysis of swallowing-related outcomes following glossectomy. AB - OBJECTIVE: To determine the effects of a partial/total glossectomy on the swallow related quality of life (QOL). DESIGN: Cross-sectional, single-centre cohort study. PATIENTS AND METHODS: Thirty-one patients who underwent partial/total glossectomy at our centre participated in the study. Main outcome was measured using the MD Andersen Dysphagia Inventory (MDADI) questionnaire. RESULTS: Responses were received from 24 males and 7 females (response rate of 77.5%) with a median age of 50 years (range: 28-72). Median follow-up in patients was 33 months (range 4-210). The mean MDADI total score in our series of patients was 71.7 (SD 18.8). Mean MDADI global score was 64.5 (SD 29.1), mean Emotional score was 71.9 (SD 19.1), mean Functional score was 73.7 (SD 19.1) and mean Physical score was 69.3 (SD 21.7). Statistically significant differences were seen between the global, emotional and physical scores of patients who had received radiotherapy (Mann-Whitney, p < 0.05) and tracheostomy (functional score, Mann Whitney, p = 0.038). CONCLUSION: The presence of tracheostomy tube and previous radiotherapy affects swallowing outcome in patients who have had surgery for squamous cell carcinoma of the tongue. PMID- 18391575 TI - Pica behavior induced by body rotation in mice. AB - OBJECTIVES: To study whether rotational stimulus induced pica and whether the vestibular apparatus was necessary for obtaining rotation-induced pica in mice. METHODS: Pica behavior in mice was investigated following 60 min of rotation once daily at 70 rpm (15 s on with 5 s off) for 3 consecutive days. After evaluating vestibular function and histology of vestibular epithelia, we examined rotation induced kaolin intake, so-called pica, in sham-lesioned and chemically labyrinthectomized mice. RESULTS: The labyrinthectomized mice exhibited loss of the contact righting and swimming capability while the destruction of hair cells of vestibular epithelia was observed. Moreover, mice subjected to rotation, but not labyrinthectomized mice, showed a significant increase in kaolin intake at the last 2 rotation sessions and the first postrotation session. CONCLUSIONS: The findings indicated that a functioning vestibular system is necessary for rotation evoking pica in mice and thus pica can be a behavioral index of motion sickness in mice. PMID- 18391576 TI - Suppression of survivin expression by short hairpin RNA induces apoptosis in human laryngeal carcinoma cells. AB - PURPOSE: The aim of this study was to evaluate the suppression effect of survivin shRNA on the expression of the survivin gene in the human laryngeal cancer cell line Hep-2. PROCEDURES: 60 cases of laryngeal squamous-cell carcinoma (LSCC) and 10 cases of normal laryngeal mucosa were examined using immunohistochemistry to determine whether the expression of survivin correlated with tumorigenesis. Three plasmid vectors of short hairpin RNA (shRNA) specific for survivin were designed and generated. Western blot and real-time PCR analysis of survivin expression in Hep-2 cells was performed 48 h after transfection. The growth curve was used to determine the cell proliferation. Propidium iodide (PI) single staining was applied to detect the cell cycle. The apoptosis of the cells was analyzed by flow cytometry with the FITC-annexin-V/PI double staining and PI single staining. RESULTS: 68.33% (41 out of 60) of tumors were positive for survivin expression and significantly associated with lymph node metastasis and advanced stage. In contrast, no expression of survivin in normal mucosa was detected. Transfection of Hep-2 cells with survivin shRNA significantly inhibited survivin expression at both the mRNA and the protein level in Hep-2 cells. Downregulation of survivin resulted in increasing the apoptosis index, but the results showed no obvious influence on cell cycle. CONCLUSIONS: This study demonstrates that survivin shRNA effectively inhibits survivin gene expression in Hep-2 cells leading to growth suppression and apoptotic induction in Hep-2 cells. PMID- 18391577 TI - Acoustic stapedial reflex in normal adults: biological behavior and determination of threshold levels. AB - OBJECTIVE: To record acoustic reflex thresholds for admittance (Y) and its components, susceptance (B) and conductance (G) in healthy subjects and to establish normative data. STUDY DESIGN: Acoustic reflex threshold was determined in 100 healthy persons, divided into 5 age groups of 20 persons each (20-29, 30 39, 40-49, 50-59 and 60-69 years old). Detection was performed for Y, B, and G at 226-, 678- and 1,000-Hz probe tones, using 500-, 1,000-, 2,000- and 4,000-Hz stimuli. RESULTS: Statistically significant differences in acoustic reflex thresholds were revealed between different components, stimuli and tones. The lowest threshold level was recorded for Y at 226 Hz using stimulus 1,000 Hz. Four percent missing values were observed at 226 Hz. Subjects belonging to different age groups had different reflex thresholds. CONCLUSION: Inclusion of acoustic reflex detection for B and G offers more detailed information about the middle ear status compared with Y alone, and appears to be sensitive in detecting subtle diseases. PMID- 18391578 TI - Incidence of sinunasal disease in laryngectomized patients. AB - OBJECTIVES: Patients having a total laryngectomy for cancer of the larynx or the hypopharynx are faced with tremendous physical and emotional challenges. Nasal airflow, which transports potentially pathogenic substances into the nose, is almost completely interrupted after laryngectomy. Postlaryngectomy patients are estimated to suffer less frequently from sinunasal diseases. The aim of this study was to evaluate and compare the incidence of sinunasal disorders in patients before and after laryngectomy retrospectively. PATIENTS AND METHODS: A questionnaire was designed comprising items addressing specific symptoms of sinunasal disease. RESULTS: The questionnaire was answered by 104 laryngectomized patients. Common colds occurred less frequently (p < 0.0001) as well as acute and chronic sinusitis (p = 0.0168) after laryngectomy. In comparison, the incidence of rhinorrhea increased significantly (p < 0.0001). Medical interventions for sinunasal disorders were rarely necessary and surgical interventions remained reserved for exceptional cases. CONCLUSION: Sinunasal diseases seem to affect laryngectomized individuals less frequently than before laryngectomy. PMID- 18391579 TI - Intravital microlesions in the human otic capsule. Detection, classification and pathogenetic significance revisited. AB - PURPOSE OF THE STUDY: To verify microcracks in human temporal bones by a method which distinguishes intravital lesions from artifacts, to revisit previous information derived from decalcified materials on their morphology and classification and to discuss the possible clinical significance. PROCEDURES: Histological analysis of 210 undecalcified adult human temporal bones bulk stained by basic fuchsin in ethanol prior to embedding in methyl methacrylate and processing by the cutting and grinding method. RESULTS: Intravital crack lesions appeared in all specimens and could be distinguished from artifacts regardless of crack morphology and size. Particularly, large microfissures were found around the labyrinthine windows, while smaller microfissures originating at the perilymphatic space were found throughout perilabyrinthine bone. Small noncommunicating fatigue microdamage was identified for the first time. All lesions were reorganized according to morphology, topography and possible clinical significance. CONCLUSION: Microfissures accumulate in the adult otic capsule as expected when bone remodeling is low. Any capsular microlesion may introduce a possible barrier in the recently discovered antiresorptive signaling pathway from inner ear structures via the lacunocanalicular osteocytic network to perilabyrinthine bone. For this reason intravital microfissures may interfere with in vivo inhibition of temporal bone remodeling and consequently offer another pathogenetic factor in otosclerosis. PMID- 18391580 TI - Effect of adenoidectomy and/or tonsillectomy on cardiac functions in children with obstructive sleep apnea. AB - BACKGROUND/AIMS: We aimed to determine the effects of adenoidectomy and/or tonsillectomy (AT) on cardiac functions in children with adenoid and/or tonsillary hypertrophy and obstructive sleep apnea syndrome (OSAS) by using echocardiography with tissue Doppler imaging facility (TDI). METHODS: Twenty-nine children with adenoid and/or tonsillary hypertrophy and OSAS and 26 children with primary snoring entered the study. Cardiac functions were assessed by echocardiography with TDI in both groups. Tests were repeated in the OSAS group 6 months after treatment with AT. RESULTS: Echocardiography showed a decrease in estimated pulmonary artery systolic pressure from 31 +/- 4.2 to 13.1 +/- 2.3 (p < 0.001). In TDI, tricuspid E(m) and E(m)/A(m) increased from 11.0 +/- 2.7 to 13.5 +/- 2.7 cm/s (p < 0.001), and 1.46 +/- 0.52 to 1.82 +/- 0.53 (p = 0.004), respectively, following AT, indicating improvement in right ventricular diastolic dysfunction. Similarly, mitral E(m) and E(m)/A(m) increased from 12.3 +/- 2.1 to 16.3 +/- 2.7 cm/s, and from 1.65 +/- 0.51 to 2.30 +/- 0.54, respectively (p < 0.001). There was no significant difference between postoperative values and control group values. CONCLUSION: TDI is a technique able to detect diastolic dysfunction unnoticeable by conventional echocardiography. Following AT, we observed improvement in both left and right ventricular diastolic functions using TDI. PMID- 18391581 TI - Renal diseases as targets of gene therapy. AB - A number of renal pathologies exist that have seen little or no improvement in treatment methods over the past 20 years. These pathologies include acute and chronic kidney diseases as well as posttransplant kidney survival and host rejection. A novel approach to treatment methodology may provide new insight to further progress our understanding of the disease and overall patient outcome. Recent advances in human genomics and gene delivery systems have opened the door to possible cures through the direct modulation of cellular genes. These techniques of gene therapy have not been extensively applied to renal pathologies, but clinical trials on other organ systems and kidney research in animal models hold promise. Techniques have employed viral and nonviral vectors to deliver gene modulating compounds directly into the cell. These vectors have the capability to replace defective alleles, express novel genes, or suppress the expression of pathogenic genes in a wide variety of kidney cell types. Focus has also been placed on ex vivo modification of kidney tissue to promote allograft survival and limit the resulting immune response to the transplanted organ. This could prove a valuable alternative to current immunosuppressive drugs and their deleterious effects on patients. While continued research and clinical trials are needed to identify a robust system of gene delivery, gene therapy techniques have great potential to treat kidney disease at the cellular level and improve patient quality of life. PMID- 18391582 TI - Nonviral gene delivery. AB - Gene and RNA interference therapies are promising cures for intractable renal failure. However, low delivery efficiency of the therapeutic nucleic acid into the nucleus of the target cell is a significant obstacle in the clinical application of nonviral gene therapy. Various mechanical techniques (hydrodynamic injection, electroporation and ultrasound-microbubble) and topically applied preparations (HVJ liposome and cationic liposome/polymer), which introduce transgenes into specific renal compartments depending on the administration route, have been reported. Additional improvements in renal application of nonviral gene vectors must address the important issue of how to control intracellular trafficking. Therefore, novel vectors based on the 'programmed packaging' concept are desirable in which all functional devices are integrated into a single system so that each function occurs at the appropriate time and correct place. In parallel with development of the carrier, quantitative evaluation of intracellular trafficking is essential to determine the efficacy of the modified devices in the cellular environment. In particular, comparison of the intracellular trafficking of the engineered devices with that of viruses (i.e. adenovirus) is useful in identifying the rate-limiting intracellular processes of the vectors during development. PMID- 18391583 TI - Retrovirus vectors. AB - In this chapter, we present an outline of retroviruses and retroviral vectors - the concepts and applications. In particular, we discuss lentiviral vectors and the suitability of these vectors for the treatment of renal pathologies. We review vector design and the data on the use of lentiviral vectors for gene transfer to the kidney. Finally, we discuss potential pathologies and avenues for the optimization of the technology for gene transfer to a complex organ such as the kidney. PMID- 18391584 TI - Adenovirus vectors for renal-targeted gene delivery. AB - Adenovirus (Ad)-based vectors are considered a promising tool for effective gene transfer in a number of renal disease-specific applications. This opinion is based upon their natural ability to infect a broad array of both dividing and terminally differentiated, nondividing cell types, their capacity to deliver (transduce) large amounts of DNA, and the ease at which this vector platform can be mass produced. Furthermore, Ads remain the gene transfer vector of choice for numerous human clinical trials, as more clinical trials utilize Ad-based vectors than any other vector currently available. However, as with all types of gene transfer vectors, several limitations to the use of Ads have been delineated. The construction of advanced-generation Ad vectors with unique modifications of the viral genome has addressed many of these issues, although continued research will no doubt provide safer alternatives to the presently used viral vectors. In this chapter, we review the current state of Ad-based gene transfer, brief updates on advanced-generation Ad-based vectors, and provide a discussion of how these vectors infect various tissues. We then specifically focus on the kidney and discuss a multitude of techniques previously employed to deliver Ad-based vectors to various regions of the kidney and in the process, reveal many associated complexities. Furthermore, exciting new studies that use Ads to express immunosuppressive gene products have shown great promise in the area of transplantation and allograft survival. Based upon this summary, we confirm that Ad-based vectors currently offer multiple advantages for the study and potential treatment of a great variety of renal and urological diseases. Utilization of advanced-generation Ad vectors combined with novel insights into the complexities of Ad-based gene transfer in general, should allow numerous inroads to be made in the near term, relative to the use of Ad-based gene transfer to treat a variety of renal diseases. PMID- 18391585 TI - Adeno-associated virus vectors: versatile tools for in vivo gene transfer. AB - Over the last few years, viral vectors based on the adeno-associated virus have gained increasing popularity due to several favorable characteristics, including the high efficiency of transduction of postmitotic tissues in vivo and the long term persistence of transgene expression in the absence of inflammation or immune response. Recently, completed trials have substantially confirmed the clinical applicability of these vectors, while indicating that further developments in vector design and production scale are necessary to broaden human application. This review summarizes our current knowledge on the molecular biology of these vectors and their clinical utilization, in particular concerning their application in renal gene transfer. PMID- 18391586 TI - RNA interference in research and therapy of renal diseases. AB - Significant improvements have been made during the last 20 years in therapy of renal diseases including the broadening of treatment options. Gene therapy is a potential modality for many renal diseases for which we are yet unable to offer specific treatment. Here, we introduce RNA interference (RNAi), one type of posttranscriptional gene silencing, as a novel gene therapeutic possibility and describe the mechanism and kinetics of action. We highlight the correlation between structure and efficacy of small interfering and short hairpin RNAs that are the most often used small RNAs possessing RNAi activity. Delivery is the biggest obstacle for RNAi-based gene therapy. Although hydrodynamic treatment is effective in animals, it cannot be used in human therapy. Possibilities to achieve site-specific and effective delivery are listed. Side effects of RNAi and potential solutions are also summarized. Besides the above-described world of small RNAs, we draw attention to the yet unrevealed function of human microRNAs that are localized mainly in the noncoding regions of the genome, are highly conserved among animals and possess important regulatory functions. Although there are many unanswered questions and problems to face in this new field of gene therapy, we summarize a number of experiments targeting renal diseases with the aid of RNAi. High specificity of short interfering RNAs and short hairpin RNAs raise hope for treating renal diseases. PMID- 18391587 TI - Gene therapy for acute renal failure. AB - Thanks to the progressive understanding of the cellular and molecular basis of renal function and disease, during the next several decades new therapeutic approaches to a wide range of kidney disorders, including acute renal failure (ARF), will be developed. In this regard, the repair of ischemic and toxic ARF is critically dependent on a redundant, interactive cytokine and growth factors network to return kidney function to near-normal baseline function. A newer strategy in biotechnology is the development of recombinant genetic engineered compounds and, recently, cell therapy derivatives. Gene therapy offers a novel approach for prevention and treatment of renal diseases. Technical advances in viral vector systems and the development of fusigenic liposome vectors have been crucial to the progress of effective gene therapy strategies directed at renal structures in animal models. Many investigations have provided experimental models for gene delivery systems but the most efficient renal gene transfer was obtained from intrarenal injection or perfusion of explanted kidneys in transplantation. Continued technologic advances in vector systems and promising results in human and animal gene transfer studies make the use of gene therapy an encouraging strategy. Cell therapy, a tool for gene therapy, is based on the ability to expand specific cells in tissue culture to perform differentiated tasks and to introduce these cells into the patient either in extracorporeal circuits or as implants as drug delivery vehicles of a single protein or to provide physiological functions. These new approaches may result in therapeutic modalities that diminish the degree of renal failure and the time needed to recover renal function in acute tubular necrosis. This article specifically examines the present prospects of gene developing therapies in the treatment of ARF. PMID- 18391588 TI - Chronic deteriorating renal function and renal fibrosis. AB - Chronic deteriorating renal function and renal fibrosis are common features in progressive renal diseases. Renal fibrosis may determine the degree of impairment of renal function and predict long-term prognosis. Advances in cell biology have provided a new understanding of the molecular events underlying renal fibrosis. A central event in tissue repair is the release of cytokines, i.e. transforming growth factor-Beta, in response to injury. The sustained expression of these cytokines underlies the development of renal fibrosis. Understanding the molecular mechanisms of the cytokines and their signaling could lead to the application of clinically useful gene therapy. PMID- 18391589 TI - Allograft rejection: acute and chronic studies. AB - Organ transplantation represents the only possible therapeutic intervention for a large number of end-stage renal diseases. The current immunosuppressive drugs are very efficient to overcome the acute rejection; however, their continuous administration exposes patients to a great risk of developing opportunistic infections and cancer. The typical drug cocktail consisting of a high-dose steroid, a calcineurin inhibitor and an anti-metabolite is effective but any of these drugs exert specific side effects including nephrotoxicity in the long term. The rate of late graft loss, essentially due to the onset of chronic allograft nephropathy, is still too excessive and can be predicted by a previous episode of acute rejection. Attempts to limit the early insults damaging the graft should exert beneficial effects on long-term graft functionality. Gene therapy, originally conceived to cure genetic diseases, has been successfully applied in the last decade to organ transplantation with the final aim to overcome acute or chronic rejection. Transfer of genes that encode proteins with immunomodulatory properties might represent a therapeutic tool to reduce and hopefully avoid the long-life administration of drugs. In this chapter, we review gene therapy studies carried out in the context of experimental organ/tissue allotransplantation to overcome acute and chronic graft rejection. PMID- 18391590 TI - Gene therapy for renal cancer. AB - Recent advances in understanding the molecular events associated with renal cell carcinoma (RCC) are revolutionizing the therapeutic options offered for patients with advanced-stage RCC. These targeted approaches for RCC are based primarily on antiangiogenesis and/or specific kinase inhibitors targeting the vascular endothelial growth factor and platelet-derived growth factor receptors, Raf and mammalian target of rapamycin inhibitor. In this context, characterization of the molecular events unique to RCC is also of critical significance for gene therapy endeavors. The attributes of gene therapy for RCC may include true targeting to cancer cells, transfer of immunomodulatory or antiangiogenic genes and novel nonapoptotic cancer cell killing mechanisms. Gene therapy may thus become a promising new adjuvant modality for RCC and expand the therapeutic armamentarium against RCC. Beyond the current stage of preclinical proof of principle and toxicological analysis in animal models, the utility of RCC gene therapy will depend on safety and efficacy trials in human subjects. These trials will determine whether targeted therapy for RCC employing genome-based strategies will broaden the current therapeutic spectrum for RCC comprising kinome-based, immunomodulatory and antiangiogenesis strategies. PMID- 18391591 TI - HIV-associated nephropathy. AB - HIV-associated nephropathy (HIVAN) is a unique form of collapsing focal segmental glomerulosclerosis that typically occurs in patients with advanced HIV disease. The pathogenesis of HIVAN involves direct HIV infection and gene expression in tubular and glomerular epithelial cells; in effect, HIVAN can be considered a natural illustration of gene delivery to the kidney. HIV infection or expression of HIV genes results in dysregulation of tubular and glomerular epithelial cells and induction of local inflammatory cascades. Specific HIV genes, in particular Nef and Vpr, play prominent and synergistic roles in the pathogenesis of HIVAN, while other viral genes are not required for the development of HIVAN. The disproportionate burden of HIVAN and HIV-related end-stage renal disease in blacks suggests that host genetic factors are also important in the pathogenesis of HIVAN. Preliminary genetic studies in the mouse model have identified a potential genetic susceptibility locus, and a number of host genes are differentially expressed in the setting of HIVAN or HIV infection. The current management of HIVAN couples antiretroviral therapy with adjunctive agents that target downstream effects of HIV gene expression in the kidney. Future therapies could also target different steps in the pathogenesis of HIVAN, including viral replication, epithelial cell entry and viral gene expression, and downstream cellular pathways. PMID- 18391593 TI - Competing causes of death for women with breast cancer and change over time from 1975 to 2003. AB - This study was to determine whether the proportion of death due to breast cancer changed over time in different cohorts of women diagnosed with breast cancer. We identified 316,149 women with breast cancer at age 20 or older during 1975-2003 from the Surveillance, Epidemiology, and End Results 9 tumor registries in the United States. Logistic regression models were used to assess the effects of time period on the likelihood of dying because of breast cancer as underlying cause of death, adjusting for other factors. Overall, underlying cause of death was 52.8% due to breast cancer, 17.8% due to heart disease, and 4.9% due to stroke. Percentage of death due to breast cancer did not change significantly from 1975 to 2003 in those who died < 12 months after diagnosis, but decreased significantly in women who died between 1 and 15 years. Risk of death due to breast cancer in women diagnosed during 1995-1998 was significantly lower than those in 1975-1979 (odds ratio = 0.79, 95% confidence interval = 0.70-0.89), after adjusting for age, race, ethnicity, and tumor stage. Percentage of death due to breast cancer decreased significantly with age from 87.5% in women < 40% to 30.7% in those 80 or older, which was not significantly affected by year of diagnosis. Proportion of death due to breast cancer increased with advanced tumor stage and was similar in various racial/ethnic groups of population. The findings demonstrated that the impact of breast cancer on overall death was reduced after 1 year of diagnosis, but suggested the need for continued cancer surveillance. PMID- 18391594 TI - High-dose chemotherapy for patients with high-risk breast cancer: a clinical and economic assessment using a quality-adjusted survival analysis. AB - OBJECTIVE: The benefit of high-dose chemotherapy (HDC) has not been clearly demonstrated. It may offer disease-free survival improvement at the expense of major toxicity and increasing cost. We evaluated the trade-offs between toxicity, relapse, and costs using a quality-adjusted time without symptoms or toxicity (Q TWiST) analysis. PATIENTS AND METHODS: The analysis was conducted in the context of a randomized trial (PEGASE 01) evaluating the benefit of HDC for 314 patients with high-risk breast cancer. A Q-TWiST analysis was first performed to compare HDC with standard chemotherapy. We then used the results of this Q-TWiST analysis to inform a cost per quality-adjusted life-year (QALY) comparison between treatments. RESULTS: Q-TWiST durations were in favor of HDC, whatever the weighting coefficients used for the analysis. This benefit was significant when the weighting coefficient related to the time spent after relapse was low (<0.38). For quite high values of this coefficient (>0.78), HDC offered no benefit. For intermediate values, the results depended on the weighting coefficient attributed to the toxicity period. The incremental cost per QALY ranged from 12,691euro/QALY to 26,439euro/QALY, according to the coefficients used to weight toxicity and relapse. CONCLUSION: The benefits of HDC outweigh the burdens of treatment for a wide range of utility coefficients. Economic impact is not a barrier to HDC diffusion in this situation. Nevertheless, no significant benefit was demonstrated for a certain range of utility values. PMID- 18391595 TI - Impact of treatment and socioeconomic status on racial disparities in survival among older women with breast cancer. AB - OBJECTIVE: To examine racial/ethnic disparities in mortality and survival in a large nationwide and population-based cohort of women with breast cancer after simultaneously controlling for differences in comorbidity, treatment, and socioeconomic status. METHODS: A cohort of 35,029 women with stage I-IIIA breast cancer at age > or = 65 from 1992 to 1999 was identified from the surveillance, epidemiology, and end results-medicare linked databases with up to 11 years of follow-up. Cox proportional hazard regression analysis was performed to determine the risk of all-cause and breast cancer-specific mortality. RESULTS: African American women with breast cancer were more likely to live in the poorest quartiles of socioeconomic status at the census tract level than whites (73.7% versus 20.7%, P < 0.001). Those living in communities with the lowest socioeconomic status were 11% more likely to die than those in the highest (hazard ratio, 1.10; 95% confidence interval, 1.04-1.16). The risk of dying changed slightly after controlling for race/ethnicity (1.11; 1.05-1.18). Compared with white women with breast cancer, crude hazard ratios of all-cause and breast cancer-specific mortality were 1.35 (1.27-1.45) and 1.83 (1.56-2.16) for African Americans. After adjusting for treatment and socioeconomic status, hazard ratio of all-cause mortality was no longer significant in African-Americans (1.02; 0.84 1.10), whereas the risk of breast cancer-specific mortality was marginally higher in African-Americans (1.21; 1.01-1.46). CONCLUSIONS: Racial disparities in overall survival between African-American and white women with breast cancer were not present after controlling for treatment and socioeconomic status. Efforts to eliminate these barriers have important public health implications for reducing disparities in health outcomes. PMID- 18391596 TI - Identification of lung cancer patients by serum protein profiling using surface enhanced laser desorption/ionization time-of-flight mass spectrometry. AB - OBJECTIVE: Diagnosis of lung cancer at an early disease stage is important for successful treatment and improving the outcome of patients. To improve its prognosis, we attempted to explore novel tools for screening serum biomarkers to distinguish lung cancer from healthy individuals by serum protein profiles and a classification tree algorithm. METHODS: Serum samples were applied to metal affinity protein chips to generate mass spectra by surface-enhanced laser desorption/ionization (SELDI) time-of-flight mass spectrometry. Protein peak identification and clustering were performed using the Biomarker Wizard software. Proteomic spectra of serum samples from 89 lung cancer patients and age- and sex matched 68 healthy individuals were used as a training set and a classification tree with 3 distinct protein masses was generated by using Biomarker Pattern software. The validity of the classification tree was then challenged with a blind test set including another 62 lung cancer patients and 34 healthy individuals. We additionally determined Cyfra21-1 and carcinoembryonic antigen in all the serum samples included in this study using an electrochemiluminescent immunoassay. RESULTS: The software identified an average of 48 mass peaks/spectrum and 3 of the identified peaks at 5808, 5971, and 7779 d were used to construct the classification tree. The classification tree separated effectively lung cancer from healthy individuals, achieving a sensitivity of 91% (81 of 89) and a specificity of 97% (66 of 68). The blind test challenged the model with a sensitivity of 89% (55 of 62) and a specificity of 91% (31 of 34), and a positive predictive value of 90% (86 of 96), respectively. The specificity of Cyfra21-1 and the sensitivity provided by Cyfra21-1 and carcinoembryonic antigen used individually or in combination were significantly lower than that of the SELDI marker pattern (P < 0.05 or P < 0.005, respectively). CONCLUSION: The results suggest that SELDI time-of-flight mass spectrometry technique can correctly distinguish lung cancer patients from healthy individuals and shows great potential for the development of a screening test for the detection of lung cancer. PMID- 18391597 TI - Lapatinib/gemcitabine and lapatinib/gemcitabine/oxaliplatin: a phase I study for advanced pancreaticobiliary cancer. AB - OBJECTIVE: To evaluate the safety/tolerability and potential antitumor activity of lapatinib, at dose ranges of 1000 to 1500 mg/d, in combination with gemcitabine and gemcitabine/oxaliplatin (GEMOX) in patients with advanced pancreaticobiliary cancer. MATERIALS AND METHODS: Patients with advanced pancreaticobiliary cancer were assigned to 1 of 4 cohorts of lapatinib administered once daily. Toxicities, response, and survival were assessed. RESULTS: Twenty-five patients were enrolled, 18 with pancreatic cancer and 7 with biliary cancer. Lapatinib, 1500 mg/d, was successfully administered with weekly gemcitabine. Dose limiting toxicities of nausea and anorexia occurred in 2 of 5 patients receiving 1500 mg/d lapatinib with GEMOX. The median survival of all patients was 11 months and the 1-year survival was 48%. CONCLUSION: Lapatinib, 1500 mg/d, can be administered with weekly gemcitabine. The maximum tolerated dose of lapatinib is 1000 mg/d with GEMOX. A phase II study of lapatinib and gemcitabine for metastatic pancreatic cancer will be initiated. PMID- 18391598 TI - PEFG (cisplatin, epirubicin, 5-fluorouracil, gemcitabine) regimen as second-line therapy in patients with progressive or recurrent pancreatic cancer after gemcitabine-containing chemotherapy. AB - OBJECTIVE: The therapeutic arsenal for salvage therapy in pancreatic cancer is limited. PEFG (cisplatin, epirubicin, 5-fluorouracil [FU], gemcitabine) regimen is an effective upfront treatment in advanced pancreatic cancer. The activity and safety of this combination regimen were assessed by means of an observational study in a population of patients with progressive or recurrent pancreatic adenocarcinoma after gemcitabine-containing chemotherapy. METHODS: Patients with age <76 years, Karnofsky performance status >50 were treated with either classic PEFG (until April 2004: cisplatin and epirubicin 40 mg/m day 1, gemcitabine 600 mg/m day 1 and 8, FU 200 mg/m/d continuous infusion day 1-28) or dose-intense PEFG (since May 2004: cisplatin and epirubicin 30 mg/m, gemcitabine 800 mg/m every 14 days; FU 200 mg/m/d continuous infusion day 1-28) until progressive disease or a maximum of 6 cycles of 28 days. RESULTS: Forty-six patients (37 metastatic) received 69 cycles of classic PEFG (18 patients) or 104 cycles of dose-intense PEFG (28 patients) as second-line therapy. Prior treatment consisted of single agent gemcitabine (N = 17), gemcitabine-based chemotherapy (N = 4), or PEFG regimen (N = 25). Median previous progression-free survival was 7.6 months. Dose intensity (mg/m/wk) with classic PEFG was cisplatin and epirubicin 8.5; gemcitabine 230; FU 1035 and with dose-intense PEFG was cisplatin and epirubicin 11.5 (+36%); gemcitabine 259 (+13%); FU 1046 (+1%). Main grade >2 toxicity consisted of neutropenia in 26 patients (56%), thrombocytopenia in 10 (22%), anemia in 11 (24%), fatigue and stomatitis in 4 (9%), vomiting, diarrhea and hand foot syndrome in 2 (4%). Partial response was observed in 11 patients (24%) (5 classic PEFG 28% + 6 dose-intense PEFG 21%). Median and 1-year survival was 8.3 months (8.0 vs. 9.0 months) and 26% (17% vs. 32%). Median and 6-months progression-free survival was 5.0 months (4.5 vs. 5.0 months) and 34% (33% vs. 38%). CONCLUSIONS: PEFG regimen in gemcitabine refractory pancreatic cancer had an acceptable toxicity profile and interesting activity, and may constitute a treatment option in this setting. PMID- 18391599 TI - Salvage chemotherapy with biweekly irinotecan, plus 5-fluorouracil and leucovorin in patients with advanced gastric cancer previously treated with fluoropyrimidine, platinum, and taxane. AB - OBJECTIVES: We retrospectively assessed the combination of biweekly irinotecan with 5-fluorouracil (5-FU), and leucovorin (LV) as salvage chemotherapy in patients with advanced gastric cancer (AGC) previously treated with fluoropyrimidine (F), platinum (P), and taxane (T). METHODS: Between October 2003 and February 2006, all 131 patients with AGC were treated with irinotecan (150 mg/m(2) on day 1), along with either FOLFIRI-1 (ie, LV (20 mg/m(2) bolus) before 5-FU (1000 mg/m(2) continuous infusion over 6-hour) on days 1-2), or FOLFIRI-2 (ie, LV (20 mg/m(2) bolus) before 5-FU (400 mg/m(2) bolus) followed by 22-hour continuous infusion of 600 mg/m(2) on days 1-2), or FOLFIRI-3 (ie, 5-FU (400 mg/m(2) bolus) followed by 46-hour continuous infusion of 2400 mg/m(2) 5-FU and 100 mg/m(2) LV). Cycles were repeated every 2 weeks. RESULTS: The median age of the patients was 52 years (range, 19-70 years). Patients received a median of 4 cycles of chemotherapy (range, 1-21 cycles). Of the 97 patients with measurable disease, 1 (1.0%) achieved a complete response, and 11 (11.3%) achieved partial responses, making the overall response rate 12.3%. The median time to progression (TTP) was 2.2 months (95% CI, 1.9-2.6 months) and the median overall survival (OS) was 6.2 months (95% CI, 5.6-6.9 months). Good performance status (P = 0.046), fewer metastatic sites (P < 0.001), and longer time to progression of previous chemotherapy (P = 0.006) were independent prognostic factors affecting OS. OS was longer with the FOLFIRI-1 regimen but not with statistical significance (P = 0.064). The treatments were generally well tolerated. CONCLUSIONS: In actual clinical practice, biweekly irinotecan with 5-FU and LV had modest activity and tolerability in AGC patients previously treated with F, P, and T. PMID- 18391600 TI - Gemcitabine plus celecoxib in patients with advanced or metastatic pancreatic adenocarcinoma: results of a phase II trial. AB - OBJECTIVES: Cycloxygenase-2 (COX-2) is overexpressed in pancreatic tumors where it may be involved in inflammation, carcinogenesis, and the regulation of neoangiogenesis. The purpose of this trial was to evaluate the combination of intravenous gemcitabine with selective COX-2 inhibitor, celecoxib for effect on survival, disease progression, and tolerability in patients with advanced pancreatic cancer. In addition, limited pharmacokinetic and pharmacodynamic analyses were preformed. MATERIALS AND METHODS: Eligible patients included those with locally advanced or metastatic pancreatic cancer with no prior chemotherapy and ECOG performance status 0-2. The treatment consisted of intravenous gemcitabine 1000 mg/m weekly x 7 weeks and concurrent daily oral celecoxib 400 mg orally twice a day. Daily oral low-dose aspirin 81 mg was administered throughout the study as a precaution for increased risk of thrombotic events. Those with stable or responsive disease were continued on intravenous gemcitabine 1000 mg/m weekly x 3 weeks and concurrent oral celecoxib. RESULTS: Twenty five patients have been enrolled at 3 centers. Five patients had locally advanced cancer; 20 had metastatic disease. The most common grade 3/4 hematological toxicities were neutropenia (32%) and anemia (20%). Four patients (17%) had partial response and 7 (35%) demonstrated stable disease. The estimated 12-month survival rate was 15%, which did not reach the predetermined efficacy end point. There was a trend suggestive of correlation between a decrease in serum vascular endothelial growth factor and patient survival. CONCLUSION: The addition of celecoxib to gemcitabine therapy did not demonstrate significant improvement in measured clinical outcomes, in patients with advanced pancreatic cancer. Higher doses of celecoxib may be needed to observe significant antitumor activity. PMID- 18391601 TI - A prospective study of quality of life in adults with newly diagnosed high-grade gliomas: comparison of patient and caregiver ratings of quality of life. AB - OBJECTIVE: To examine whether a caregiver can provide reliable proxy quality of life (QOL) ratings of their adult significant other with a newly diagnosed high grade glioma. METHODS: This prospective QOL study was a companion protocol for 3 phase II high-grade glioma protocols. At study entry, 2 months, and 4 months after enrollment, 5 self-administered forms were completed by 197 patients and their caregivers to assess QOL. RESULTS: Caregiver ratings of QOL were available, respectively, for 92%, 93%, and 88% of baseline, 1st, and 2nd subsequent follow up evaluations of patients who had completed their QOL assessments. There was a strong relationship between patient and caregiver QOL scores (Spearman and intraclass correlation coefficients greater than 0.5 for 87% and 80% of the measurements, respectively); however, for some measures (eg, the profiles of mood states short form) there was better agreement between patient and caregiver scores when the QOL scores were higher. There was good agreement between patient and proxy ratings independent of the cognitive function of the patient, except for the profiles of mood states short form with better correlation between patients and caregivers for those patients without cognitive impairment. CONCLUSIONS: In this multi-institutional prospective study there is a strong correlation between high-grade glioma patient and caregiver QOL scores, although for some measures this correlation is stronger for those patients without cognitive impairment. To improve the acquisition and the accuracy of assessing QOL status in high-grade glioma patients, proxy ratings from caregivers should also be obtained in conjunction with the patient, and consideration be given to substituting proxy ratings when a patient's self-report is absent. PMID- 18391602 TI - Radiographic complete response on post treatment CT imaging eliminates the need for adjuvant neck dissection after treatment for node positive head and neck cancer. AB - OBJECTIVE: The primary purpose of this study was to correlate radiographic response in the neck to clinical outcomes for patients with node positive head and neck cancer. METHODS: One hundred three patients with stage III/IV node positive cancer were treated with definitive radiotherapy or chemoradiation at a single institution from 1990 to 2004. Follow-up ranged from 8 months to 144 months with a median of 42 months. Posttreatment CT scans were called complete radiographic response (rCR) or partial radiographic response. RESULTS: Actuarial 36 month rates of survival, control above the clavicles, and nodal control were 66%, 74%, and 90%, respectively. Patients, who had rCR on posttreatment CT scan, who had a neck dissection had a nodal control rate of 94% compared with those without neck dissection of 97%. Patients with partial radiographic response who were treated with neck dissection had a nodal control rate of 94% compared with those without neck dissection of 73%. CONCLUSIONS: Based on this data, there was no suggestion that neck dissection improved outcome for patients with rCR on posttreatment imaging 4 to 6 weeks after radiation. PMID- 18391603 TI - Autologous large multivalent immunogen vaccine in patients with metastatic melanoma and renal cell carcinoma. AB - OBJECTIVE: To evaluate the safety and activity of large multivalent immunogen (LMI), prepared by immobilizing autologous tumor cell plasma membrane on 5-microm diameter silica beads, in patients with melanoma and renal cell carcinoma (RCC). METHODS: Thirty patients with stage IV metastatic melanoma and 31 patients with stage IV RCC were randomly assigned to 1 of 3 trial arms and received monthly treatment with (1) LMI alone, (2) cyclophosphamide followed 8 days later with LMI, or (3) the same treatment as in arm 2 with IL-2 given for 5 days beginning 1 week after LMI administration. RESULTS: No grade 4 toxicities were observed. For patients with melanoma, median overall survival time for all 30 patients was 20.4 months [95% confidence interval (CI): 8.0-not assessable], and median progression free survival was 2.8 months (95% CI: 1.9-6.3). For patients with RCC, median overall survival exceeded 46.2 months (95% CI: 30.3-not assessable), and median progression-free survival was 12.2 months (95% CI: 4.6-not assessable). Two patients had a partial response to LMI treatment. CONCLUSIONS: Based on our results that demonstrate the safety and tolerability of LMI vaccine, further development of this therapy is warranted to evaluate its clinical efficacy. PMID- 18391604 TI - Sorafenib for metastatic renal cancer: the Princess Margaret experience. AB - OBJECTIVE: Sorafenib, an oral multikinase inhibitor, prolonged progression-free survival when compared with placebo, as second-line therapy for patients with metastatic renal carcinoma (MRC). Grade 3/4 adverse events were reported in 12% of patients. This study presents sorafenib's efficacy and safety in a less selected cohort of patients enrolled in an expanded access program. METHODS: Patients with MRC received sorafenib 400 mg twice daily until disease progression. Tumor response was evaluated by RECIST criteria. Adverse events were graded by NCI common toxicity criteria. RESULTS: From November 2005 to August 2006, 58 patients were enrolled. The median progression-free survival was 7.5 months (95% CI: 5.4-11.3), and the best responses among 54 patients were 11 (20%) confirmed partial responses, 15 (28%) stable diseases for > or =6 months; 10 patients (18%) had early progression at 8 weeks. Grade 3/4 adverse events occurred in 37 patients (64%; 95% CI: 50%-76%), the most frequent being skin rash in 17 patients (29%), and hand-foot syndrome in 9 patients (15%). Thirty-six (62%) patients required dose reductions and/or treatment interruptions. CONCLUSIONS: Sorafenib is effective in a less selected patient population with MRC but leads to more toxicity than described previously. PMID- 18391605 TI - Anticancer effect of combination therapy of VP16 and fosfesterol in hormone refractory prostate cancer. AB - OBJECTIVES: We conducted the present study to evaluate the safety profile and therapeutic value of a combination of etoposide and fosfestrol for treatment of hormone-refractory prostate cancer (HRPC). METHODS: Forty patients with HRPC were included in the study. The median age was 71 years (range, 50-86 years), the Gleason's score ranged from 5 to 10, and the median prostate-specific antigen level was 62.6 ng/mL (range, 4.738-30789 ng/mL). The patients received oral etoposide 25 mg/d and fosfestrol 300 mg/d. RESULTS: The response rate in terms of measurable disease, serum prostate-specific antigen level, and overall evaluation was 36.8% (CR: 18.4%; PR: 18.4%), 80% (CR: 55%; PR: 25%), and 40% (CR: 20%; PR: 20%) with a median duration of response of 13.6, 13.5, and 13.5 months, respectively. An objective clinical response for overall evaluation was shown by 90% (CR: 20%; PR: 20%; SD: 50%) of the patients, with a median response duration of 15.7 months; 16 patients (40%) are currently alive without recurrence after a median follow-up period of 21.2 months. The overall survival and progression-free survival was 30.5% and 28.8% at 40 months, respectively. No grade III toxicities occurred in any of the patients. Serial measurements in 34 patients using the Functional Assessment of Cancer Therapy-Prostate showed a significant improvement in quality of life as a result of the therapy. CONCLUSIONS: The combination of oral etoposide and fosfestrol is active in patients with HRPC. The regimen is tolerable and has a significant impact on quality of life as measured by the Functional Assessment of Cancer Therapy-Prostate in a limited sample of patients. PMID- 18391606 TI - Sphincter preservation for distal rectal cancer: how low is too low? PMID- 18391607 TI - Fistula formation after postoperative radiation treatment for paranasal sinus cancer. AB - Postoperative radiation is frequently used in the treatment paradigm for paranasal sinus tumors. The development of 3-dimensional conformal radiation treatment and intensity modulated radiotherapy (IMRT) has facilitated the delivery of high doses required for local control of these lesions while simultaneously decreasing toxicity. At Memorial Sloan-Kettering Cancer Center, a radiation dose of 70 Gy is routinely prescribed to gross tumor, and 59.4 Gy is prescribed to a clinical target volume at high risk for subclinical disease and 54 Gy is delivered to a clinical target volume at low risk for subclinical disease. Fistula formation can occur with the delivery of postoperative radiation treatment despite the use of IMRT. Prosthesis fabrication can be used in the short-term management of this unfortunate complication with an acceptable cosmetic result. Patients should be aware of this potential toxicity, which can develop in spite of appropriate management and acceptable dosimetry. Nonetheless, combined modality therapy is recommended for aggressive treatment of paranasal sinus tumors to inhibit local progression. This report describes the clinical scenario and management of the rare incidence of fistula formation after radiation for paranasal sinus malignancy. PMID- 18391608 TI - Metastatic synovial sarcoma of the foot to the throat. PMID- 18391609 TI - Primary liver sarcoma in an immunocompetent patient with hepatitis C virus infection. PMID- 18391610 TI - Methods of epidemiology: evaluating the fat-breast cancer hypothesis--comparing dietary instruments and other developments. AB - Results from several large cohort studies that were reported 10 to 20 years ago seemed to indicate that the hypothesized link between dietary fat intake and breast cancer risk was illusory. In this article, we review several strands of more recent evidence that have emerged. These include two studies comparing the performance of dietary instruments used to investigate the dietary fat- breast cancer hypothesis, a large randomized disease prevention trial, a more recent meta-analysis of nutritional cohort studies, and a very large nutritional cohort study. Each of the studies discussed in this article suggests that a modest but real association between fat intake and breast cancer is likely. If the association is causative, it would have important implications for public health strategies in reducing breast cancer incidence. The evidence is not yet conclusive, but additional follow-up in the randomized trial, as well as efforts to improve dietary assessment methodology for cohort studies, may be sufficient to provide a convincing answer. PMID- 18391611 TI - Erythropoietin-stimulating agents in oncology. AB - Erythropoiesis stimulating agents (ESAs) are some of the most widely used agents in oncology. Yet, the use of ESAs to treat chemotherapy induced anemia in cancer patients has raised issues of concern since 2003, when the two initial studies that were powered to detect meaningful differences in survival with ESA therapy were either halted early, or concluded with deleterious effects on survival. Several trials since then have caused both guideline writers and the FDA to recommend that ESAs should not employed to drive the hemoglobin to greater than 12 g/dL. It is still unclear what role these agents will play in cancer patients with hemoglobin of less than 12 g/dL. In myelodysplastic syndrome, however, these agents are likely part of a core of supportive care measures for low-grade disease, and need to be readily available for these patients. In this review, we attempt to describe the relevant experimental data on ESAs, their current role in clinical oncology, and the research and discoveries that may widen the scope, and enhance the benefit of these agents. PMID- 18391612 TI - Chemotherapy-induced nausea and vomiting. AB - Chemotherapy-induced nausea and vomiting (CINV) affects many cancer patients and has a great influence on quality of life. CINV involves coordination of several organs of the gastrointestinal tract, the peripheral and central nervous systems. Many neurotransmitters are involved in this process, and the predominant receptors are serotonin, neurokinin-1 and dopamine receptors. Risk factors for CINV include patient gender and age, past history of CINV, plus the emetogenicity and administration schedule of chemotherapy. Recommended antiemetic regimens for highly emetogenic chemotherapy and moderately emetogenic chemotherapy with a high risk of delayed CINV include a serotonin antagonist, dexamethasone and aprepitant. Other moderately emetogenic chemotherapy requires a serotonin antagonist and dexamethasone. Medications for breakthrough symptoms include dopamine antagonists, lorazepam, metoclopramide, haloperidol, droperidol and other agents. Options for treatment of refractory CINV include olanzapine, dronabinol, nabilone, gabapentin. New evidence from non-controlled studies supports the use of olanzapine, casopitant and gabapentin in controlling the symptoms of CINV. PMID- 18391613 TI - From the guest editors. Hepatocellular carcinoma. PMID- 18391614 TI - Liver transplantation for hepatocellular carcinoma. AB - Liver transplantation has emerged as an optimal treatment for stage I and II hepatocellular carcinoma for patients with underlying cirrhosis as it provides a treatment for the underlying liver disease as well as a reduced incidence of recurrent cancer. The current system of organ allocation in the United States allows an opportunity for liver transplantation for patients with tumor burden within the Milan criteria (a single tumor 2-5 cm or up to 3 lesions with none >3 cm). Outcomes of patients receiving transplants within these criteria approach outcomes for patients receiving transplants for all indications (85.9%, 74.8%, and 64.1% actuarial survival at 1, 3, and 5 years, respectively, for those with HCC receiving transplants compared with 82%, 73%, and 67% for the entire cohort). Transarterial chemoembolization, radiofrequency ablation, and other pretransplant treatment modalities aimed to slowing tumor growth for patients on a transplant waiting list are commonly used, although the impact on pretransplant disease progression or posttransplant survival remains uncertain. There is continued controversy over expanding patient selection criteria, in particular for those who have undergone downstaging of tumors. In addition, the role of certain immunosuppressive agents such as sirolimus in the reducing HCC recurrence posttransplant remains unclear. PMID- 18391615 TI - Surgical resection of hepatocellular carcinoma. AB - The incidence of hepatocellular carcinoma (HCC) is increasing in the United States, primarily due to hepatitis C-related liver disease. Nearly 85%-90% of patients with HCC have underlying chronic liver disease or cirrhosis. Advanced tumor burden or prohibitive hepatic dysfunction precludes operative resection in most patients with HCC. Surgical resection is a treatment option with curative intent in patients with HCC not associated with cirrhosis or in patients with well-compensated liver disease. Tumor extent and hepatic function must be assessed preoperatively to avoid postresection hepatic failure, an often fatal condition that may require urgent liver transplantation. Appropriately selected candidates for liver resection have 5-year postoperative survival rates of 40% 70%, but recurrence rates approach 70%, especially in patients with cirrhosis. For this reason, the best resection for patients with HCC and cirrhosis is orthotopic liver transplantation, which has 5-year posttransplant survival rates of 65%-80% in well-selected candidates. PMID- 18391616 TI - Hepatocellular carcinoma: radiation therapy. AB - Although whole liver tolerance to radiation therapy (RT) is low, hepatocellular carcinoma (HCC) can be treated with focal high-dose RT, using a variety of advanced and specialized treatment strategies. Technical advancements in external beam RT that facilitate the safe delivery of RT to a wide spectrum of patients include conformal RT planning, breathing motion management, and image-guided RT. A variety of doses and RT fractionation schemes have been used safely alone or in combination with other therapies such as transarterial chemoembolization. Charged particles, produced from very specialized treatment units, are associated with particularly desirable dose distributions allowing tumoricidal doses to be delivered with sustained tumor control and little toxicity, even in the presence of Child-Pugh class B or C cirrhosis. Another strategy to deliver RT to HCC is hepatic arterial delivery of radioisotopes, such as microspheres tagged with yttrium-90. Liver toxicity is more likely in patients with reduced liver reserve and/or tumors infiltrating the majority of the liver. Phase II studies and a small phase III trial have demonstrated activity of hepatic arterial radioisotopes in HCC, providing rationale for large confirmatory randomized trials. Recurrences after RT occur most often within the liver, outside the high dose irradiated volume, and outcomes after RT to very large and/or diffuse HCC are poor, providing rationale for combining RT with other therapies or novel radiation sensitizers. Given the vascular properties of HCC, there is rationale for investigating RT with anti-vascular endothelial growth factor-targeted agents. PMID- 18391617 TI - Radiofrequency ablation and chemoembolization for hepatocellular carcinoma. AB - PURPOSE: To provide an up-to-date review of the technique, efficacy, safety and clinical applications for radiofrequency ablation (RFA) and transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC). METHODS: A thorough review of the literature was performed as related to RFA and TACE for the treatment of HCC. We mixed these findings with our own extensive research and clinical experience on this subject by drawing from a pool of more than 1000 patients treated with both modalities. RESULTS: TACE has been shown with the work of Llovet, Camma, and Lo to provide a significant survival benefit for patients with unresectable HCC. The former showed a 1-, 2-, and 3-year survival at 57%, 31%, and 26% in the treatment group vs. 32%, 11%, and 3% in the supportive treatment alone group, respectively. Repeatedly, RFA has been shown to be very effective in the treatment of small (or=3 of the following criteria: increased waist circumference (>88 cm or 35 inches), hypertriglyceridemia (>or=150 mg/dL), low (<40 mg/dL) high-density lipoprotein, hypertension, and high fasting glucose (>or=110 mg/dL). RESULTS: : One hundred-seven RA and 85 SLE patients with a mean age of 43 +/- 13 years were included in this study. The frequency of obesity and abnormal waist circumference were similar in RA and SLE patients. Two percent were underweight, 35% had a normal weight, 37% were overweight, and 25% were obese. The frequency of metabolic syndrome in RA and SLE patients was 17%. Metabolic syndrome was significantly associated with greater age, less education, lower income, and smoking. In RA patients, metabolic syndrome was significantly associated with a shorter treatment period with methotrexate, with pain, and with health assessment questionnaire scores. By multivariate logistic regression, the only statistically significant predictor of metabolic syndrome was smoking. CONCLUSIONS: The frequency of metabolic syndrome in RA and SLE patients was similar and associated with smoking. In RA patients, metabolic syndrome was related with pain and functional status, suggesting disease activity. A better control of disease activity may reduce the presence of metabolic syndrome and the risk of cardiovascular disease. PMID- 18391675 TI - ANCA vasculitis in the elderly. AB - BACKGROUND: In population-based estimates of disease, the incidence of ANCA associated vasculitides is increasing, with the peak age of onset in middle aged and older adults. Clinical characteristics of the very elderly have not been fully elucidated. METHODS: Patients who met the criteria for Wegener granulomatosis and microscopic polyangiitis were included in the study. The following characteristics were analyzed by retrospective review: time interval from first symptoms to date of diagnosis, age at diagnosis, laboratory values on initial presentation, and Birmingham Vasculitis Activity Score when diagnosed. RESULTS: Patients >75 years old were more often female, had fewer ear, nose, or throat symptoms, and had lower hemoglobin values. At date of latest follow-up, 40% of the older patients died compared with 11% of the younger cohort (P = 0.0006). Of the deceased elderly patients, nearly one-half died within 6 months. Age >75 carried an elevated risk of mortality (hazard ratio = 2.69, 95% confidence interval = 1.55-4.96; P = 0.0005) as did elevated serum creatinine (hazard ratio = 1.25 per 1 mg/dL, 95% confidence interval = 1.06-1.43; P = 0.0109). Survival was worse in those presenting with Birmingham Vasculitis Activity Score >20 and those >75-year-old. CONCLUSION: The clinical presentation of ANCA-associated vasculitis is relatively similar among elderly and younger patients. In patients aged 75 years or older, ANCA vasculitis is associated with higher mortality, and related to the presence of renal involvement. Elderly patients have a greater risk for death within the first 6 months after diagnosis. PMID- 18391676 TI - Trochanteric bursitis: refuting the myth of inflammation. AB - BACKGROUND AND OBJECTIVES: Greater trochanteric (GT) bursitis is a common cause of hip pain. Previously, the etiology of the trochanteric pain syndrome was thought to be caused by inflammation of the subgluteus maximus bursa (i.e., bursitis). Recently, MRI and ultrasound studies have brought into serious doubt the idea that bursitis is the etiology for trochanteric pain. To our knowledge, no histologic study of GT bursitis has been reported to date. The purpose of this study is to evaluate the histopathology of patients with and without the clinical syndrome of GT bursitis to assess for the presence of bursal inflammation. DESIGN AND METHODS: This is a prospective, case-controlled, blinded study of the histopathologic features of controls and patients with GT bursitis. We recruited patients who required total hip arthroplasty (THA) for rheumatoid or osteoarthritis. Inclusion criteria for the study consisted of the following: needing THA as standard of care; THA secondary to OA or RA; age greater than 18; and minimal risk for surgery by the American Heart Association Criteria. We excluded anyone who received a GT bursa injection 9 months before surgery. Eligible participants were then stratified as cases or controls using the 1985 clinical criteria for GT bursitis. The harvesting of the bursa required no modification of the surgical procedure. The specimens were then examined by 2 independent pathologists who were blinded as to the patients' clinical status. RESULTS: Six bursal specimens were evaluated by 2 blinded surgical pathologists revealing primarily fibroadipose tissue with no signs of acute or chronic inflammation. There were 3 bursas in the control group and 2 specimens with clinical GT bursitis. No significant differences were found between the specimens of the 2 groups. CONCLUSIONS: The results of this small prospective observational histologic study, along with recent MRI and ultrasound studies on the topic, strongly suggest that there is no etiologic role of bursal inflammation in the trochanteric pain syndrome. PMID- 18391677 TI - The sensitivity and specificity of the Slump and the Straight Leg Raising tests in patients with lumbar disc herniation. AB - BACKGROUND: An accurate and specific diagnosis prevents the recurrences of low back pain and chronic spinal pain. The physical examination is the most useful tool to diagnosis. The examiner must aim to determine the exact tissue that pain arises from to make the specific diagnosis. Lumbar disc herniation is 1 disease that physical examination, symptoms, and findings on imaging technique do not always correlate with each other. The Straight Leg Raising (SLR) test has been used as the primary test to diagnosis lumbar disc herniations and found to have high correlation with findings on operation since its sensitivity is high in only disc herniations leading to root compression that may eventually need operation. More sensitive test, like the Slump, might be used in herniations in which the SLR is negative. The Slump test is really a variant of the SLR and the Lasegue's tests performed in the seated position and is a progressive series of maneuvers designed to place the sciatic nerve roots under increasing tension. At each step in the procedure, the patient informs the examiner what is being felt and whether radicular pain is produced. As a result, the Slump test applies traction to the nerve roots by incorporating spinal and hip joint flexion into the leg raising and would warn the examiner of the presence of nerve root compression when there is a negative SLR test. OBJECTIVES: This study measured the sensitivity and specificity of the Slump test and compare it with the SLR test in patients with and without lumbar disc herniations. METHODS: A prospective case control study of 75 patients with complaints suggestive of lumbar disc herniation was carried out in the outpatient clinics of the neurosurgery department of a state teaching hospital. Seventy-five referred or self-admitted patients with low back, leg, or low back and leg pain who had results of magnetic resonance imaging (MRI) of the lumbar spine were included in the study. Thirty-eight patients had signs of herniation demonstrated by MRI. Control patients (n = 37) had no disc bulges or herniations on MRI. Both the Slump and SLR tests were performed during the assessment of all the patients by the second author. The MRI results were assessed and recorded by the first author. RESULTS: The Slump test was found to be more sensitive (0.84) than the SLR (0.52) in the patients with lumbar disc herniations. However, the SLR was found to be a slightly more specific test (0.89) than the Slump test (0.83). CONCLUSION: The Slump test might be used more frequently as a sensitive physical examination tool in patients with symptoms of lumbar disc herniations. In contrast, owing to its higher specificity, the SLR test may especially help identify patients who have herniations with root compression requiring surgery. PMID- 18391678 TI - Effect of rituximab on refractory Wegener granulomatosis with predominant granulomatous disease. AB - Wegener granulomatosis is an autoimmune disorder with a double spectrum. Vasculitic manifestations are at one end of it, whereas granulomatous ones are at the other. Rituximab (RTX) is a chimeric monoclonal antibody that has been successfully used in this condition. However, the granulomatous forms have been reported to show a tendency to be less responsive to RTX than the vasculitic disease. We present 4 cases of predominantly Wegener granulomatosis that responded positively to RTX. PMID- 18391679 TI - Successful remission of thrombotic thrombocytopenic purpura with rituximab in a patient with undifferentiated connective tissue disorder. AB - Thrombotic thrombocytopenic purpura (TTP) is rarely associated with undifferentiated connective tissue disorder. We present a patient with TTP, undifferentiated connective tissue disorder, and very high titer of anti ribonucleoprotein antibodies. Her TTP did not respond to intravenous methylprednisolone and cyclophosphamide requiring her to remain dependent on plasmapheresis. Her disease remitted successfully after 4 doses of rituximab given at weekly intervals and she remained in continuous remission 6 months after therapy. We propose early and aggressive use of B-cell depletion therapy in TTP associated with autoimmune disorders. PMID- 18391680 TI - A rare case of enterobacter endocarditis superimposed on a mitral valve rheumatoid nodule. AB - We present the case of a 56-year-old man with longstanding seropositive active erosive and deforming rheumatoid arthritis with no peripheral rheumatoid nodules; he immigrated from the former Soviet Union (where he did not receive any disease modifying antirheumatic drugs) to Israel in 1995. In February 2005, he had a buccogingival mucosal abscess on his lower lip, which was treated by surgical drainage, followed by prolonged antibiotic therapy. One and a half years later, he had 2 episodes of transient ischemic attacks characterized by speech difficulties and moderate weakness on his right side. Transesophageal echocardiogram revealed a mass on the anterior mitral valve leaflet. Repeated blood cultures were negative, and the patient was afebrile all the time. The patient underwent mitral valve replacement and the histologic findings of the mass were typical of both a rheumatoid nodule and bacterial endocarditis. The patient recovered fully after 6 weeks of antibiotic therapy. Emboli from a rheumatoid nodule should always be considered in patients with rheumatoid arthritis who present with transient ischemic attacks. PMID- 18391681 TI - Normal initial magnetic resonance imaging in aseptic bone necrosis of the knee. AB - Magnetic resonance imaging (MRI) is considered the most sensitive tool for the detection of osteonecrosis at early stages. We report about 3 patients with high suspicion of osteonecrosis at the knees in whom an initial MRI was normal, but the condition was apparent in a subsequent study. The time between the initial insult and the appearance of diagnostic alterations in MR studies remains undefined and, in patients at risk an initial MRI, if done too early, may fail to detect this condition. PMID- 18391683 TI - Clinician's corner--advocates for the rheumatologist--practice, treatment, and education. PMID- 18391684 TI - Pediatric rheumatology for the adult rheumatologist. AB - Although many manifestations of rheumatic diseases in children are similar to those in adults, there exist important differences. These include variations in presentation and complications, differential diagnoses to consider, medication side effects, impact on growth, and psychosocial issues. As there is a US and worldwide shortage of pediatric rheumatologists, many children with rheumatic illnesses are cared for by other physicians, especially by adult rheumatologists. This article is aimed toward educating the adult rheumatologist in 4 key areas of childhood rheumatic disease: juvenile idiopathic arthritis, other rheumatic and inflammatory illnesses seen primarily in the pediatric population, differences in rheumatic diseases common to both adults and children, and psychosocial issues. PMID- 18391685 TI - Synovial fluid findings in a patient after hylan GF 20 injection. PMID- 18391686 TI - Rheumatoid arthritis masked by docetaxel chemotherapy in a patient with ovarian carcinoma. PMID- 18391687 TI - Prior bisphosphonate therapy of osteoporosis attenuates and blocks response to subsequent parathyroid hormone. PMID- 18391688 TI - Swollen hand as presenting manifestation of syringomyelia. PMID- 18391689 TI - Failed intubation in a patient with long-standing ankylosing spondylitis. PMID- 18391690 TI - Onset of psoriasis induced by infliximab. PMID- 18391691 TI - Psoriasis induced by anti-TNF probably not so uncommon. PMID- 18391692 TI - Anti-TNF agents inducing psoriasis: a recognized adverse effect. PMID- 18391693 TI - Post-thrombotic syndrome in children: a single center experience. AB - BACKGROUND: Development of post-thrombotic syndrome (PTS) is increasingly being recognized as a complication of deep venous thrombosis (DVT) in children. OBJECTIVE: To determine the prevalence, clinical characteristics, and predictors of moderate to severe PTS in children. METHODS: A retrospective chart review was performed on those children who were followed in the coagulation clinic for objectively confirmed DVTs from December 2004 to December 2006. The scoring system used by Kuhle et al was used to grade the severity of PTS as: mild, moderate, and severe. RESULTS: PTS developed in 20% (11/55; 95% confidence interval 9.4-30.1) of children, in which 8/11 were moderate and 3/11 were severe. Median interval between diagnosis of PTS and DVT was 90 days (range, 46 d to 3 y). The majority (72.7%) of patients in the non-PTS group received treatment intervention within 48 hours of diagnosis of DVT. Delay in treatment initiation (>48 h) and recurrence of DVT were associated with the development of PTS (P<0.05). Variables including occlusive thrombus, location and number of vessels involved with DVT, age at diagnosis, underlying thrombophilia, intensity of anticoagulation, and body mass index were not associated with the development of PTS. Other debilitating consequences of DVT requiring intervention included portal hypertension (n=2), chylothorax (n=1), and reflux sympathetic dystrophy (n=1). The small sample size and limited follow up restricted the statistical analysis. CONCLUSIONS: PTS is a significant problem in children with symptomatic DVTs. Early treatment intervention within the first 48 hours of diagnosis of DVT and prevention of thrombosis recurrence may prevent development of PTS. Although PTS refers to consequences of intravenous hypertension owing to extremity DVTs, sequlae of nonextremity DVTs require special consideration in pediatric PTS classification. PMID- 18391694 TI - SFCE (Societe Francaise de Lutte contre les Cancers et Leucemies de l'Enfant et de l'Adolescent) recommendations for the management of tumor lysis syndrome (TLS) with rasburicase: an observational survey. AB - Rasburicase (Fasturtec), a recombinant urate oxidase, is highly effective in preventing and treating hyperuricemia in children with hematologic malignancies. We conducted a prospective, multicenter observational study in 174 patients at 8 pediatric hemato-oncology centers to establish whether the SFCE (Societe Francaise de Lutte contre les Cancers et Leucemies de l'Enfant et de l'Adolescent) recommendations for the use of rasburicase in the management of pediatric patients at risk of tumor lysis syndrome (TLS) are valid in routine clinical practice. Patients were classified as being at high or low risk of TLS according to the Children's Oncology Group criteria and were treated in accordance with the SFCE recommendations. The primary end point was the number of patients requiring a higher dose of rasburicase or a longer duration of treatment than advised in the SFCE recommendations. Of the 135 patients at high risk of TLS, 27 patients received a higher dose and 35 patients received a longer duration of treatment. Some patients received treatment with rasburicase for less than the recommended duration (median 4 d for high-risk patients). One patient required hemodialysis. Only minor adjustments to the SFCE recommendations were required to ensure the optimal use of rasburicase in pediatric patients at risk of TLS. PMID- 18391695 TI - A community-based physical activity program for adolescents with cancer (project TREK): program feasibility and preliminary findings. AB - INTRODUCTION: As the number of pediatric survivors continues to grow, a greater emphasis is being placed on identifying long-term health risk/protecting behaviors. The primary purpose of this pilot study was to examine the feasibility of a theoretically-based physical activity (PA) intervention in adolescents with cancer. METHODS: A group PA intervention was administered to participants over 16 weeks. Program attendance/adherence, total PA, physical fitness, and quality of life (QOL) were assessed at 5 different intervals over the 1-year study duration. RESULTS: Over a period of 6 weeks, 11 adolescents responded to recruitment efforts. Ten ultimately enrolled and completed the 1-year study. Overall, the program was well received with attendance over the 16-week intervention averaging 81.5%. Although improvements in total PA, physical fitness, and QOL were noted across the intervention, follow-up data revealed that participants failed to maintain their postintervention PA levels at both the 3 and 12-month follow-up assessments. CONCLUSIONS: Adolescents with a history of cancer are undoubtedly susceptible to the same maladaptive health habits as are their healthy peers. Innovative interventions aimed at improving their adherence to positive behavioral interventions, such as PA are warranted. Although not without limitations, this study provides preliminary data in support of a university sponsored PA intervention. PMID- 18391696 TI - Correction of aberrant pre-mRNA splicing by antisense oligonucleotides in beta thalassemia Egyptian patients with IVSI-110 mutation. AB - The splicing mutation in intron 1 of beta-globin gene (IVS1-110) is the most common mutation in Egyptian thalassemics that causes aberrant splicing of pre mRNA and deficient beta-globin chain synthesis. Antisense oligonucleotides (ASONs) are compounds that redirect pre-mRNA splicing and modify gene expression. Our aim was ex vivo correction of the aberrant splicing of beta-globin110 pre mRNA by ASON against the 3' aberrant splice site. Peripheral blood mononuclear cells of 10 thalassemic patients with IVS1-110 mutation were duplicated and 1 was treated with 20 micromoL/mL morpholino ASON targeted against the 3' aberrant splice site. The level of total hemoglobin (Hb), fetal Hb, and mRNA were estimated in the duplicate samples. Five cases (50%) showed correction with ASON treatment, of which 2 cases showed the appearance of corrected mRNA band with absence of the aberrant band and 3 cases showed an increased ratio of the corrected to the aberrant mRNA band from 2:1 to 3:1, and 4:1. The total Hb showed significant increase in the 5 corrected cases. In conclusion, ASON can restore correct splicing of beta-globin pre-mRNA leading to correct gene product in cultured erythropoietic cells. These results suggest the applicability of ASON for the treatment of thalassemia. PMID- 18391697 TI - Pediatric Hodgkin lymphoma in 2 South American series: a distinctive epidemiologic pattern and lack of association of Epstein-Barr virus with clinical outcome. AB - Hodgkin lymphoma (HL) shows a bimodal distribution with a first peak in developing countries during childhood. The causative role and prognostic significance of Epstein-Barr virus (EBV) association in patients with HL is controversial. Our aim was to perform a comparative study of EBV association in 2 Latin American pediatric HL series, and to correlate it with patient's survival. Epstein-Barr encoded RNAs in situ hybridization and latent membrane protein 1 immunohistochemistry were performed on formalin-fixed, paraffin-embedded HL biopsies from 176 pediatric patients from 2 public institutions from Argentina and Southeast Brazil. Mixed cellularity subtype was prevalent in Argentine HL (Arg HL) (52%) and nodular sclerosis subtype in Brazilian HL (BR HL) (83%). EBV expression was detected in 52% of cases, namely 54% Arg HL and 48% Br HL. EBV was significantly associated with mixed cellularity subtype in both populations. In Arg HL, EBV positivity was significantly higher in patients0.05) under all loading conditions. The TLIF procedure significantly reduced the range of motion compared with the intact state (P=0.03). However, no statistical difference was detected between the TLIF group and the ALIF group (P>0.05). CONCLUSIONS: Both ALIF and TLIF procedures combined with posterior instrumentation significantly improved construct stability of intact spinal motion segments. However, there was no statistical difference between these 2 fusion techniques. The 2 ALIF approaches (lateral and anterior) also had similar construct stability even though anterior longitudinal ligament severing significantly reduced stability. PMID- 18391718 TI - Prevalence of lumbar total disc replacement candidates in a community-based spinal surgery practice. AB - STUDY DESIGN: This study was a retrospective record review of consecutive lumbar surgical patients seen by one surgeon in a private, community setting over a 15 month period. OBJECTIVE: The purpose of this study was to evaluate the potential frequency that candidates for total disc replacement (TDR) might be seen in a community-based spinal surgery practice. SUMMARY OF BACKGROUND DATA: The impact of TDR technology on the practice of spine surgery remains uncertain. The only previous report in the literature evaluating prevalence of contraindications to TDR reflected a patient cohort from a high volume, academic, tertiary referral center, and may not accurately reflect the experiences of the "average" spine surgeon, in community practice. Further, this study did not assess the prevalence of patients who possessed an indication for TDR. METHODS: We performed a retrospective chart review of consecutive patients who underwent lumbar spine surgery between 2003 and 2004 (during which period the senior author was in a community-based private practice). Particular attention was paid to conditions considered to be contraindications to TDR. The patients were divided into nonfusion and fusion subgroups. The percentage of patients with exclusion criteria was calculated in each group, as was the percentage of potential TDR candidates. RESULTS: A total of 190 patients were identified: 124 had nonfusion procedures and 66 had fusion procedures. In the nonfusion group, all patients had at least 1 contraindication to TDR. In the fusion group, 86% of the patients had at least 1 contraindication to TDR. Of those fusion patients with degenerative disc disease, 33% had no contraindication to TDR, and were considered potential candidates for TDR. Seventeen percent of all fusion patients had Worker's Compensation Insurance; among those patients considered to be potential TDR candidates, 44% had Worker's Compensation Insurance, and a further 22% of TDR candidates were on permanent disability (Medicare) as a result of a work-related injury. CONCLUSIONS: This study suggests that in a community-based spinal surgery practice setting, only 14% of fusion patients, and only 5% of all lumbar surgery patients, would be TDR candidates. Furthermore, of those potential TDR candidates, as many as 66% might be expected to have their underlying condition related to a claim of a compensable work-related injury. PMID- 18391719 TI - Increasing bending strength and pullout strength in conical pedicle screws: biomechanical tests and finite element analyses. AB - STUDY DESIGN: Comparative in vitro biomechanical study and finite element analysis. OBJECTIVES: To investigate the bending strength and pullout strength of conical pedicle screws, as compared with conventional cylindrical screws. SUMMARY OF BACKGROUND DATA: Transpedicle screw fixation, the gold standard of spinal fixation, is threatened by screw failure. Conical screws can resist screw breakage and loosening. However, biomechanical studies of bending strength have been lacking, and the results of pullout studies have varied widely. METHODS: Ten types of pedicle screws with different patterns of core tapering and core diameter were specially manufactured with good control of all other design factors. The stiffness, yielding strength, and fatigue life of the pedicle screws were assessed by cantilever bending tests using high-molecular-weight polyethylene. The pullout strength was assessed by pullout tests using polyurethane foam. Concurrently, 3-dimensional finite element models simulating these mechanical tests were created, and the results were correlated to those of the mechanical tests. RESULTS: In bending tests, conical screws had substantially higher stiffness, yielding strength, and fatigue life than cylindrical screws (P<0.01), especially when there was no step at the thread-shank junction. In pullout tests, pullout strength was higher in screws with a conical core and smaller core diameter and also in situations with higher foam density (P<0.01). In finite element analysis, the maximal deflection and maximal tensile stress were closely related to yielding strength (r=-0.91) and fatigue life (r=-0.95), respectively, in the bending analyses. The total reaction force was closely related to the pullout strength in pullout analyses (r=0.84 and 0.91 for different foam densities). CONCLUSIONS: Conical screws effectively increased the bending strength and pullout strength simultaneously. The finite element analyses reliably predicted the results of the mechanical tests. PMID- 18391720 TI - Success of lumbar microdiscectomy in patients with modic changes and low-back pain: a prospective pilot study. AB - STUDY DESIGN: Prospective case controlled. OBJECTIVE: To determine the outcome after microdiscectomy in patients with disc herniation, concordant sciatica, and low-back pain with Modic I and II degenerative changes compared with similar patients without Modic changes. SUMMARY OF BACKGROUND DATA: The decision to perform a microdiscectomy versus a fusion or total disc replacement in a patient with a disc herniation and sciatica may be confounded by the presence of low-back pain, degenerative disc disease, and marrow and endplate (Modic) changes. METHODS: Thirty consecutive patients underwent a microdiscectomy by a single surgeon. Group 1 consisted of 15 patients, 6 men and 9 women, with a mean age of 36.7 years (range, 21 to 48 y), with Modic I and II changes. Group 2 contained 15 patients, 9 men and 6 women, with a mean age of 34.1 years (range, 20 to 68 y), without Modic changes. The average duration of low-back pain before surgery was 25.6 months (range 4 to 120 mo) in group 1 and 17.5 months (range 5 to 120 mo) in group 2. The visual analog scale (VAS) was used to grade low-back pain and the Oswestry score was used to grade overall disability. RESULTS: There was no significant difference in preoperative sciatica, low-back pain, VAS or Oswestry scores for group 1 versus group 2 patients. Postoperatively, all patents had improved sciatica and resolution of any nerve tension sign. Eighty-six percent of patients in group 1 versus 93% of patients in group 2 had improvements in postoperative VAS score for low-back pain at 6 months. Average improvement within each group was 67% and 75%, respectively. VAS scores for low-back pain at 6 months improved from 6.9 to 2.3 (P=0.0005) in group 1 and 6.3 to 1.6 (P=0.0002) in group 2. Group 1 and 2 had 89% and 100% of patients show improvement in postoperative Oswestry score at 6 months with an average improvement of 58% and 84%, respectively. Oswestry scores at 6 months improved from 68.7% to 28.8% (P=0.0007) in group 1 and 61.2% to 9.9% (P=0.00003) in group 2. CONCLUSIONS: There was a trend toward greater improvement in Oswestry scores in patients without Modic changes (P=0.09). Both groups reported statistically significant improvement in sciatica, low-back pain, and disability after microdiscectomy. Microdiscectomy was therefore an effective treatment for disc herniation and concordant sciatica despite low-back pain and Modic I and II degenerative changes. LEVELS OF EVIDENCE: Therapeutic II. PMID- 18391721 TI - Analysis of interleukin-8, interleukin-10, and tumor necrosis factor-alpha in the cerebrospinal fluid of patients with cervical spondylotic myelopathy. AB - STUDY DESIGN: Retrospective cohort study. OBJECTIVE: The expression of interleukin-8 (IL-8), IL-10, and tumor necrosis factor-alpha (TNF-alpha) were measured in the cerebrospinal fluid (CSF) of patients with cervical myelopathy. The purpose of this study was to examine whether the CSF levels of those 3 cytokines differ significantly among 3 groups of patients with different diseases. METHODS: IL-8, IL-10, and TNF-alpha levels were analyzed using enzyme linked immune assay. CSF samples were collected from 3 groups of patients. The cervical spondylotic myelopathy (CSM) group consisted of 35 patients. The ossification of the posterior longitudinal ligament group [(OPLL) group] consisted of 7 patients, and the control group consisted of 12 patients. The concentration of IL-8 was 69.0+/-35.2 pg/mL in the CSM group, 82.1+/-46.7 pg/mL in the OPLL group, and 43.5+/-20.9 pg/mL in the control group. The concentration of IL-8 was significantly higher in the CSM and OPLL groups than in the control group (P<0.05). There was no significant difference between the CSM group and OPLL group. The concentration of IL-10 and TNF-alpha in all groups was below the sensitivity of the measurements. CONCLUSIONS: In this study, the concentration of IL-8 was high in CSM and OPLL patients. However, the concentration of IL-10 and TNF-alpha was below the sensitivity of the measurements. PMID- 18391722 TI - The CSF creatine kinase-BB isoenzyme activity in experimental lumbar spinal stenosis model. AB - STUDY DESIGN: An investigation of creatine kinase (CK)-BB isoenzyme activity in cerebrospinal fluid (CSF) of the rabbits after experimentally induced spinal stenosis. OBJECTIVES: To create a lumbar spinal stenosis (LSS) model at conus medullaris level without laminectomy in rabbits and to investigate the importance of CK-BB isoenzyme activity in CSF associated with electrophysiologic and histopathologic changes in the spinal cord. SUMMARY OF BACKGROUND DATA: LSS is a disorder characterized by leg pain and difficulty of walking. Narrowing of the spinal canal and compression on the spinal cord and nerves are the main features of spinal stenosis. METHODS: Fifteen male albino rabbits were used in this study. A reproducible, subacute LSS model was created in all rabbits, and CSF CK-BB activity was measured above and below the stenosis level. The electrophysiologic evaluation and the histopathologic examination of the conus medullaris were also performed in each rabbit. RESULTS: The CK-BB activity was 71.5% in CSF samples that were obtained below the stenosis. The activity was 44.5% in samples obtained above the stenosis and 43.6% in nonstenotic rabbits. In the electrophysiologic studies, the mean amplitudes were decreased and the latency values were lengthened in all ascending and descending nerve potentials at both sides of the stenosis. The number of the neural cells was decreased and imperception of the nucleolus of neural cells and vacuolar degeneration were observed in the histopathologic examination of conus medullaris. CONCLUSIONS: The activity of CK BB isoenzyme was increased in CSF of which the circulation was disturbed as a result of neural ischemia, which was accepted in the pathophysiology of LSS. PMID- 18391723 TI - 68Ga-DOTA-peptides versus 18F-DOPA PET for the assessment of NET patients. PMID- 18391724 TI - Guidelines for the use of PET-CT in children. PMID- 18391725 TI - Optimizing delayed scan time for FDG PET: comparison of the early and late delayed scan. AB - OBJECTIVES: Although dual-time-point scans have been widely used to improve the diagnostic efficacy of FDG PET in differentiating between malignant and benign lesions, no optimized delayed scan time-point has yet been recommended in clinical practice. Our study aimed to explore the most appropriate time for a delayed scan by comparing early and late delayed scans. METHODS: Eighty patients with suspected malignancy were given a three-phase (64 min, 110 min, 233 min after FDG injection) PET/CT scan. The maximum standardized uptake values (SUVs) in the three-phase scans were recorded as SUV1, SUV2 and SUV3, respectively, and compared among three-phase imaging. Retention indices (RIs) of each lesion in two delayed phases were calculated according to the formulae: RI1=SUV2-SUV1/SUV1 x100% and RI2=SUV3-SUV1/SUV1 x100%. RI1 and RI2 in both malignant and benign groups were assessed through correlation analysis. The diagnostic values of two delayed scans were compared through the analysis of the receiver operating characteristic curves. RESULTS: One hundred and nine of 148 lesions were malignant, and 39/148 lesions benign, which were verified by pathological, clinical, laboratory or radiological examination. RI1 and RI2 in malignancy were 14.8+/-13.1% and 10.8+/-20.5% respectively, and the correlation coefficient was 0.6 (P=0.0001). RI1 and RI2 in benign lesions were 11.3+/-28.2% and 9.3+/-42.4%, respectively, and the correlation coefficient was 0.6 (P=0.0001). The area under the ROC curve for RI1 was 0.627+/-0.050 (null hypothesis: true area=0.5, P=0.0130); whereas the area under the ROC curve for RI2 was 0.563+/-0.052 (null hypothesis: true area=0.5, P=0.2321), suggesting that the late delayed scan may have no diagnostic value. CONCLUSION: The retention index values in the two delayed phases have good relativity. The diagnostic value of early delayed imaging is higher than that of late delayed imaging. An early delayed scan, according to our research, should be recommended in clinical practice. PMID- 18391726 TI - Prognostic value of FDG uptake by the bone marrow in squamous cell carcinoma of the head and neck. AB - BACKGROUND: The appearance of natural suppressor cells and circulating endothelial progenitor cells in tumour tissue has been associated with myelopoetic stimulation by growth factors that may increase fluorodeoxyglucose (FDG) uptake by the bone marrow and high FDG uptake by bone marrow in patients suffering from human malignancies is a not uncommon finding. METHODS: This study looked at the relationship between bone marrow FDG uptake, biochemical (Hb level, RBC count, WBC count and platelet count), clinical and radiological findings and outcome in a series of 35 patients suffering from squamous cell carcinoma of the head and neck (SCCHN), consecutively referred for FDG PET as part of their routine staging procedure. RESULTS AND CONCLUSION: In SCCHN, mean FDG standardized uptake values (SUVs) of the primary tumour correlate significantly with blood WBC count (r=0.44; P=0.011, Bonferroni corrected P=0.04) and mean FDG SUVs of bone marrow are significantly correlated to the maximum FDG SUVs of the primary tumour (r=0.523; P=0.002). Finally, FDG uptake by the bone marrow is related to disease-free and overall survival. These findings warrant confirmation in a larger patient series. PMID- 18391727 TI - The role of 99mTc-depreotide in the management of neuroendocrine tumours. AB - BACKGROUND: In-pentetreotide scan (OctreoScan) is a widely available agent with high sensitivity for imaging neuroendocrine tumours. Negative In-pentetreotide poses diagnostic as well as therapeutic problems in terms of staging and consideration of targeted radionuclide therapy. AIM: To assess the role of Tc depreotide in patients with negative or weakly positive OctreoScan (Krenning score< or =1; measured on a scale range 0-4). To determine the usefulness of Tc depreotide scintigraphy for highlighting lesions that may be missed by OctreoScan and/or CT/MRI imaging. STUDY DESIGN: Prospective analysis of 25 patients with neuroendocrine tumours, with negative or weakly positive In-pentetreotide scans, who were consecutively enrolled to undergo In-pentetreotide and Tc-depreotide imaging. The results were compared with either CT or MRI scans. RESULTS: Histology was available for 20 of 25 patients: of these 40% had high-grade tumours (cellular proliferation marker Ki-67 score >20%), a further 35% had intermediate-grade tumours (Ki-67 2-20%), and the remaining 25% had low-grade tumours (Ki-67 <2%). Fifty-two percent of patients had completely negative and 48% had weakly positive OctreoScan results. Thirty-two percent of these same patients had significantly positive Tc-depreotide scans (Krenning score> or =2), with the histology demonstrating intermediate-grade or high-grade tumours. CONCLUSION: Tc-depreotide imaging has low sensitivity but is useful in a one third of OctreoScan-negative patients, displaying significantly better uptake than In-pentetreotide in this patient group. It aids diagnosis by highlighting lesions not seen by OctreoScan and/or CT/MRI imaging, and can possibly identify a group of patients amenable to therapy with radionuclide agents, such as SOM230, targeting somatostatin receptor subtypes 2, 3 and 5. PMID- 18391728 TI - Changes in the appearance of attenuation artefacts due to change in posture in myocardial perfusion imaging. AB - OBJECTIVE: Traditional supine imaging with arms raised for myocardial perfusion imaging (MPI) is uncomfortable for many cardiac patients. Seated imaging with arms resting at shoulder level is an attractive alternative. This study aimed to compare the patient comfort and image appearance of seated MPI with traditional supine MPI. METHOD: Sixty-seven patients (41 male, 26 female; body mass index (BMI) between 20.4 and 45.4) were imaged seated on the Mediso Nucline Cardiodesk gamma camera and supine on the GE Millennium VG gamma camera using our standard departmental MPI protocol. The images from each were compared and a questionnaire was used to determine patients' views of the relative comfort of the procedures. RESULTS: Strong patient preference for seated imaging was demonstrated. Perfusion patterns on seated and supine images were strikingly different with the changes seen being greatest and very striking in obese females with seven out of 12 (58%) showing changes of more than two grades. For a subset of 17 normal weight (BMI <25) male patients a significant reduction (P<0.05) in defect size was found in the inferior segment only, signifying a reduction in diaphragmatic attenuation in the seated position. CONCLUSION: Seated imaging offers considerable advantages in terms of patient acceptability. For non-obese men seated imaging also offers advantages in terms of reduced diaphragmatic attenuation artefacts. However, women and also obese men show significant differences in perfusion pattern from traditional supine imaging. A facility for accurate attenuation correction of seated images could provide useful information to elucidate these effects. PMID- 18391729 TI - Myocardial scar and insulin resistance predict cardiovascular events in severe ischaemic myocardial dysfunction: a perfusion-metabolism positron emission tomography study. AB - BACKGROUND: Clinical outcome can be predicted by metabolism-perfusion positron emission tomography (PET) in patients with severe ischaemic left ventricular dysfunction. This study determined whether the amount of viable or non-viable myocardium detected with a PET scan or clinical-functional parameters might predict cardiovascular events. METHODS: All patients had previous myocardial infarction (>6 months previously) and left ventricular ejection fraction (LVEF) <40%. Metabolism-perfusion PET, echocardiogram and coronary angiography were provided. All subjects underwent short euglycaemic-hyperinsulinaemic clamp before the metabolism study. The dysfunctioning segment was defined as hibernating myocardium when metabolism was normal-moderately reduced with impaired perfusion (mismatch flow-metabolism). Cardiac death, hospital admission for myocardial infarction or heart failure were considered cardiovascular events. RESULTS: Ninety-three patients (71 males, aged 64.2 years) were studied. The LVEF was 30.2+/-7.7%; 48 (51.6%) suffered an anterior myocardial infarction. Fifty-three (54.1%) subjects were treated with coronary revascularization; all had optimal medical therapy. Cardiovascular events occurred in 20/93 patients at 1-year follow-up (event group). Age (P=0.7), diabetes mellitus (P=0.6) and rate of coronary revascularization (P=0.3) were not different in the two groups. Patients who experienced cardiovascular events had larger non-viable myocardium (5.8+/-2.7 vs. 4.1+/-2.6, P=0.01), lower metabolic rate glucose (1.3+/-0.6 vs. 1.7+/-0.7 ml . kg . min, P=0.04) but similar hibernating myocardium (1.6+/-1.6 vs. 1.7+/-2, P=0.8) and baseline LVEF (28.1+/-4.8 vs. 30.7+/-8.3%, P=0.08). Having more then five non-viable segments and a metabolic rate for glucose of <0.9 mg . kg . min predicted a worse prognosis (P=0.04, log rank, 3.89; and P=0.004, log rank, 8.1, respectively). CONCLUSION: Non-viable myocardium revealed with PET predicts mid term clinical prognosis. Insulin resistance seems to influence the outcome. PMID- 18391730 TI - In-vivo PET imaging of implanted human retinal pigment epithelium cells in a Parkinson's disease rat model. AB - BACKGROUND AND AIM: Researchers find that monitoring the differentiation of implanted cells in vivo is difficult. This study was designed to show that it is possible to track the efficacy of transplanted human retinal pigment epithelial cells (RPE cells) in a rat model of Parkinson's disease by using positron emission tomography (PET). METHODS: RPE cells or normal saline were injected into striatum of the injured side of the rat model in treated and control groups, respectively. PET imaging of both groups was undertaken before transplantation and at intervals afterwards, using C-raclopride and C-beta-CFT as the markers. Observation of the rats' behaviour and immunofluorescence confocal microscopy were also used to prove the PET results. RESULTS: PET studies showed increased accumulation of C-raclopride and decreased C-beta-CFT in the injured side of striatum in both groups. C-raclopride decreased along with a concomitant increase of C-beta-CFT after transplantation in the treated group. The changes shown by the PET studies paralleled the behavioural states and confocal microscopy observations in the treated animals. CONCLUSION: These results suggest that even a clinical PET scanner could, to a certain extent, provide some information on the existence and in-vivo differentiation of RPE cells in a rat model of Parkinson's disease. PMID- 18391731 TI - Radiation field feature of 188Re esophageal stent. AB - OBJECTIVE: To evaluate the radiation field feature of Re esophageal stent and provide scientific basis for clinical application. METHODS: We measure the beta ray, gamma-ray and bremssfrahlung dose of every selected point on the bionics esophageal stent and then draw out computer software by mathematical formula. RESULTS: The radiation field of Re esophageal stent has its own feature: the max range of beta-ray is 11 mm, 90% dose construction field is within 1.5 mm, 95% dose range is within 2.5 mm, and only 4.21% of total energy of gamma-ray and bremssfrahlung is out of 6.5 mm range. The absorption dose of every direction in same point of the esophageal model was similar (P>0.05). CONCLUSION: Beta ray is the major radiation of Re esophageal stent while gamma-ray and bremssfrahlung are 4.21% among the radiation field. The max dose construction field is within 0.5 1.5 mm, just short at the depth of esophagus mucosa within 0.5-1.5 mm range. So Re stent is a good choice of palliative intracavitary radiotherapy of esophageal carcinoma. PMID- 18391739 TI - Acute myocardial infarction due to a bee sting manifested with ST wave elevation after hospital admission. PMID- 18391740 TI - Gastric zygomycosis: unusual cause of gastric perforation in an immunocompetent patient. PMID- 18391741 TI - Methicillin-resistant Staphylococcus aureus: fatal prostatic abscess in an AIDS patient. PMID- 18391742 TI - Benign esophageal schwannoma. PMID- 18391743 TI - Response to "Predictors of endoscopic and laboratory evaluation of iron deficiency anemia in hospitalized patients". PMID- 18391745 TI - Histologic distinction between subungual lentigo and melanoma. AB - The distinction between a benign subungual pigmented macule (lentigo) and an early lesion of melanoma in situ can be difficult. To identify histologic parameters of potential diagnostic value, we retrospectively reviewed biopsies and excisions of 35 pigmented nail lesions. We studied 20 melanomas (10 invasive and 10 noninvasive) and 15 benign subungual melanotic lentigines. Ten specimens of normal nail apparatus obtained for reasons other than melanonychia were also examined as controls. The parameters, which were analyzed, included the density of melanocytes, the presence of multinucleated cells, pagetoid spread, cytologic atypia, inflammation, and the distribution of melanin pigment. The density of melanocytes was measured as the number of cells per 1 mm stretch of subungual dermo-epithelial junction [=melanocyte count (MC)]. The MC for invasive melanomas was as follows: mean=102, median=92.5, and range 52 to 212. For noninvasive (only in situ) melanoma, the mean MC was 58.9, median 51, and range 39 to 136. For benign subungual melanotic macules, the mean MC was 15.3, median 14, and range 5 to 31. In normal controls, the mean MC was 7.7, median 7.5, and range 4 to 9. Qualitative features associated with in situ melanoma and useful for its distinction from benign subungual melanotic macules included the presence of confluent stretches of solitary units of melanocytes, multinucleated melanocytes, lichenoid inflammatory reaction, and florid pagetoid spread of melanocytes. PMID- 18391746 TI - NUT rearrangement in undifferentiated carcinomas of the upper aerodigestive tract. AB - BACKGROUND: Undifferentiated carcinomas of the upper aerodigestive tract (UCUAT) occur most frequently within the nasopharynx and are most often associated with infection by Epstein-Barr virus (EBV) (WHO undifferentiated nonkeratinizing squamous cell carcinoma). An unusual group of aggressive carcinomas are characterized by translocations that involve Nuclear Protein in Testis (NUT), a novel gene on chromosome 15. In about two-thirds of cases, NUT is fused to BRD4 on chromosome 19. These tumors, here termed NUT midline carcinomas (NMCs), are undifferentiated, may have focal squamous differentiation, and are reported to occur in children and young adults. This study investigates the prevalence of NUT rearrangement and the diagnostic significance of NUT expression in a series of upper aerodigestive tract undifferentiated carcinomas. The histologic features of these tumors are described in detail. METHODS: All UCUAT not associated with EBV infection seen at the University of Virginia (UVA) over a 16-year period were reviewed. Clinical and histologic features were noted. Additional material was submitted for fluorescent in situ hybridization (FISH) using split-apart probes to the NUT and BRD4 genes. Immunohistochemistry (IHC) was performed on all cases using a polyclonal antibody to NUT, and on select cases with antibody to p63. RESULTS: Thirty-one UCUAT were identified. Twenty-five tumors had originally been diagnosed as sinonasal undifferentiated carcinomas. Five of 28 cases (2 males, 3 females; average age 47; range 31 to 78) with interpretable results showed rearrangements of the NUT and BRD4 genes by FISH. Three of these 5 cases showed diffuse (>90%) nuclear staining for NUT by IHC; 22 of 23 other tumors showed at most focal (<50%) nuclear staining. Undifferentiated carcinomas with NUT gene rearrangement had focal abrupt squamous differentiation in 2 cases, and intense and diffuse immunoreactivity with antibody to p63 in 4 cases. CONCLUSIONS: Approximately, 20% of UCUAT not associated with EBV infection were found to have rearrangements of NUT by FISH. Although previous reports suggest that NMCs afflict only children and young adults, 4 of 5 of the patients described are mature adults older than any heretofore reported, suggesting that previous reports may have been biased in their case selections. Furthermore, because these tumors are indistinguishable from other poorly differentiated carcinomas, IHC using NUT antibody may be a useful method for the identification of these tumors. Despite the lack of overt squamous differentiation in most cases, their p63 immunoreactivity suggests that NMCs may generally be of squamous lineage. PMID- 18391747 TI - Lung adenocarcinoma: modification of the 2004 WHO mixed subtype to include the major histologic subtype suggests correlations between papillary and micropapillary adenocarcinoma subtypes, EGFR mutations and gene expression analysis. AB - The histologic heterogeneity of lung adenocarcinoma creates a variety of complex challenges to pathologists in analyzing the various subtypes. Current classification schemas do not take into account the major subtype. We analyzed 100 cases for clinical, pathologic, and molecular features using a modification of the 2004 World Health Organization (WHO) classification to record the major component in the mixed subtype tumors. The tumors were analyzed for KRAS mutation and epidermal growth factor receptor (EGFR) by mutation, chromogenic in situ hybridization, and immunohistochemistry for EGFR and phosphorylated EGFR. Gene expression analysis was performed using HG-U133A Affymetrix oligonucleotide microarrays revealing 3 gene clusters. The most common major histologic subtype was papillary (37%) followed by acinar (30%), solid (25%) and bronchioloalveolar (7%) carcinoma (BAC), although no pure BACs were seen. Sixteen tumors harbored EGFR mutations and 14 KRAS mutations. Papillary adenocarcinoma strongly correlated with EGFR mutation (P<0.001) and gene profile Cluster 1 (P=0.006) with weaker correlations with low grade (P=0.038) and favorable behavior in Stage 1 patients (P=0.047). Micropapillary subtype correlated strongly with EGFR mutation (P<0.001) and weakly with Cluster 1 (P=0.030). Solid adenocarcinoma strongly correlated with gene profile Cluster 3 (P=0.001) and worse survival (P=0.001). BAC as a major component strongly correlated with gene Cluster 2 (P=0.001). Cluster 1 correlated strongly with less smoking (P<0.001), papillary histology (P<0.001), and EGFR mutations (P<0.001). Cluster 3 strongly correlated with heavier smoking (P<0.001), larger tumor size (P<0.001), solid subtype (P<0.001), and poor grade (P=0.004); weak correlations were found with KRAS mutation (P=0.025). No correlation was found with BAC and EGFR by mutation, chromogenic in situ hybridization or immunohistochemistry. Higher stage (P<0.001), grade (P<0.001), and solid subtype (P=0.001) correlated with shorter survival. Our data suggest that EGFR mutations are associated with papillary adenocarcinoma and gene profile Cluster 1. We discovered this only because we used a comprehensive approach examining in detail all histologic subtypes and we modified the 2004 WHO mixed subtype to include the major histologic subtype. As we do not know the major genetic pathways of 30% to 70% of lung adenocarcinomas, the comprehensive histologic subtyping we propose gives advantage for recognition of unanticipated histologic-genetic correlations that might not be detected using classification systems that focus primarily on specific aspects of adenocarcinomas such as BAC or EGFR mutations. Such an approach should be considered in future studies for validation in other datasets. PMID- 18391748 TI - Risk-reducing total gastrectomy for germline mutations in E-cadherin (CDH1): pathologic findings with clinical implications. AB - Hereditary diffuse gastric cancer is a rare autosomal dominant cancer susceptibility syndrome caused by germline E-cadherin (CDH1) mutations in 40% of cases with a high degree of penetrance. Screening endoscopy has not been useful in identifying early cancer, in part owing to conflicting data concerning site(s) of involvement in the stomach and the lack of endoscopically detectable pathology. Risk-reducing total gastrectomy specimens from 8 asymptomatic adults with germline mutations in the CDH1 gene (3 different pedigrees) were studied using a sequential serial sectioning protocol with submission of the entire stomach for histologic analysis. The presence, size, and distribution of signet ring cell clusters were determined for each section and geographic maps of the invasive foci were constructed and compared with gastrectomy specimens from patients with germline E-cadherin mutation and symptomatic gastric cancer. All but 1 of the asymptomatic patients with germline mutations in the CDH1 gene had negative endoscopic screening. All risk-reducing gastrectomy specimens were macroscopically normal. All contained multiple foci (mean, 10.9) of microscopic intramucosal signet ring cell carcinoma confined to the superficial gastric mucosa; no invasion of submucosa was identified. In situ carcinoma was present in 6/8 cases. The majority of signet ring foci were located in the proximal one third of the stomach, most within oxyntic-type mucosa. The number and size of foci were not related to age, but there was a trend toward more severe disease burden in women. Stomachs from the symptomatic group of patients with germline CDH1 mutations exhibited infiltrative foci with higher Ki-67 labeling that extended well beyond the superficial mucosa. In addition, while superficial signet ring cancer exhibited decreased or absent E-cadherin and beta-catenin protein expression in all cases studied, deeply invasive signet ring cancer showed reversion to E-cadherin and beta-catenin protein expression in a subset of mutation carriers. Our study indicates that superficial intramucosal signet ring carcinoma, although widespread, is predominantly located in the proximal one third of the stomach in patients with E-cadherin gene mutations. The observed site predilection suggests a possible role for geographically targeted endoscopic surveillance biopsy in patients who elect to delay surgical intervention. PMID- 18391749 TI - Malignant hepatic angiomyolipoma: report of a case and review of literature. AB - Hepatic angiomyolipoma, a constituent of the group of tumors showing differentiation resembling perivascular epithelioid cells, is primarily appreciated in its benign form. Regardless of their location, this family of tumors is characterized by the presence of mature adipocytes, blood vessels, and spindle-epithelioid cells. These tumors also possess similar immunohistochemical profiles, including positivity for melanocytic (HMB-45) and smooth muscle (smooth muscle actin) markers. Here, we present a case of malignant hepatic perivascular epithelioid cells that was initially confined to the liver, treated by resection, and subsequently recurred within and metastasized beyond the liver. We take this opportunity to report the fourth case of malignant hepatic angiomyolipoma (HAML) and review the literature. We will discuss the features that aid in distinguishing between benign and malignant HAML, and their similarities. In summary, the common features of both benign and malignant HAML include the following: the 3 basic histologic components of AML, expression of melanocytic and smooth muscle markers, invasion into adjacent normal parenchyma, and cytologic atypia. The unique features of malignant HAML are as follows: clinical evidence of aggressive behavior such as metastasis or death owing to disease, coagulative necrosis, and loss of CD 117 expression. PMID- 18391750 TI - The HVTN protocol 903 vaccine preparedness study: lessons learned in preparation for HIV vaccine efficacy trials. AB - Successful recruitment and retention of HIV-uninfected at-risk participants are essential for HIV vaccine efficacy trials. A multicountry vaccine preparedness study was started in 2003 to assess enrollment and retention of HIV-negative high risk participants and to assess their willingness to participate in future vaccine efficacy trials. HIV-negative high-risk adults were recruited in the Caribbean, in Southern Africa, and in Latin America, and were followed for 1 year. Participants included men who have sex with men, heterosexual men and women, and female sex workers. History of sexually transmitted infections and sexual risk behaviors were recorded with HIV testing at 0, 6, and 12 months, and willingness to participate in future vaccine trials was recorded at 0 and 12 months. Recruitment, retention, and willingness to participate in future trials were excellent at 3 of the 6 sites, with consistent declines in risk behaviors across cohorts over time. Although not powered to measure seroincidence, HIV seroincidence rates per 100 person-years (95% confidence interval [CI]) were as follows: 2.3 (95% CI: 0.3 to 8.2) in Botswana, 0.5 (95% CI: 0 to 2.9) in the Dominican Republic, and 3.1 (95% CI: 1.1 to 6.8) in Peru. The HIV Vaccine Trials Network 903 study helped to develop clinical trial site capacity, with a focus on recruitment and retention of high-risk women in the Americas, and improved network and site expertise about large-scale HIV vaccine efficacy trials. PMID- 18391751 TI - Host genetic influences on highly active antiretroviral therapy efficacy and AIDS free survival. AB - We studied the influence of AIDS restriction genes (ARGs) CCR5-Delta32, CCR2-64I, SDF1-3'A, IL10-5'A, CX3CR1-V249I, CX3CR1-T280M, and MDR1-C3435T and haplotypes of the CCR5 P1 promoter and RANTES variants -403A, In1.1C, 3'222C, and -28G among HIV-1 infected patients on highly active antiretroviral therapy (HAART) in the Multicenter AIDS Cohort Study (MACS) and the Multicenter Hemophilia Cohort Study (MHCS). Our results indicate that several ARGs also influence therapy efficacy (ie, the success in viral suppression) and subsequent progression to AIDS while on HAART. CCR5-Delta32 decreased time to viral suppression (<200 HIV RNA copies/mL, relative hazard [RH]=1.40; P=0.008) and was protective against AIDS (RH=0.11; P=or<0.0001), whereas the CCR5 P1 haplotype was associated with delayed viral suppression (RNA<50 copies/mL, odds ratio [OR]=0.65; P=0.03) and accelerated time to AIDS (RH=2.68; P=0.02). SDF1-3'A reduced viral suppression (OR=0.61; P=0.02) and accelerated AIDS (RH=3.18; P=0.009). Accelerated AIDS progression was also observed with the RANTES haplotype carrying RANTES-IN1.1C and RANTES-3'222C (P=0.005 to 0.007). In contrast, the RANTES haplotype H1, which lacks suspected deleterious single-nucleotide polymorphisms, was protective against AIDS. CX3CR1-V249I seemed to accelerate viral suppression (RNA<50 copies/mL, OR=1.27; P=0.01). ARG influence after HAART suggests residual HIV-1 replication, and spread continues even in patients successfully suppressing detectable viral RNA. PMID- 18391752 TI - Role of cytotoxic T-lymphocyte-mediated immune selection in a dominant human leukocyte antigen-B8-restricted cytotoxic T-lymphocyte epitope in Nef. AB - OBJECTIVES: To study the role of cytotoxic T-lymphocyte (CTL) escape for disease progression in HIV-1 infection, we analyzed the CTL response to the dominant human leukocyte antigen (HLA)-B8-restricted CTL epitope FLKEKGGL (FL8) in HIV-1 Nef. METHODS: HIV-1 nef genes derived from 56 patients were analyzed by polymerase chain reaction (PCR)-based sequencing. T-cell responses against FL8 and mutated FL8 variants were detected by gamma-interferon (gamma-IFN) enzyme linked immunospot (ELISPOT) assay. RESULTS: The longitudinal analysis of an HIV-1 infected patient with good control of HIV-1 viremia for several years demonstrated an association of rising viremia with the emergence of CTL escape mutations within the HLA-B8-restricted Nef-specific CTL epitopes FLKEKGGL and WPAIRERM. Analysis of nef genes in 56 HIV-1-infected patients demonstrated a significant correlation between the occurrence of mutations in the FL8 epitope and the presence of HLA-B8. The mutations within the FL8 epitope could decrease CTL recognition; however, there was strong variation regarding the recognition of viral variants between individual donors. The presence of FL8 mutations was associated with lower CD4 cell counts and higher viral loads. CONCLUSIONS: Our data demonstrate a strong CTL selection pressure on the immunodominant HLA-B8 restricted CTL epitope FL8 in HIV-1 Nef. The association of FL8 mutations with lower CD4 cell counts indicates an important role of CTL escape mutations for disease progression. PMID- 18391753 TI - A multipeptide vaccine is safe and elicits T-cell responses in participants with advanced stage ovarian cancer. AB - Nine participants with epithelial ovarian, fallopian tube, or primary peritoneal carcinoma, who were human leukocyte antigen (HLA)-A1, HLA-A2, or HLA-A3, were eligible to enroll in a phase 1 study designed to assess the safety and immunogenicity of a peptide-based vaccine. Participants received 5 class I major histocompatibility complex-restricted synthetic peptides derived from multiple ovarian cancer-associated proteins plus a class II major histocompatibility complex-restricted synthetic helper peptide derived from tetanus toxoid protein. The vaccines were administered with granulocyte macrophage-colony stimulating factor in Montanide ISA-51 adjuvant over a 7-week period. All vaccine-related toxicities were grade 1 to 2, the most common being injection site reaction (grade 2, 100%), fatigue (grade 1, 78%), and headache (grade 1, 67%). Lymphocytes from the peripheral blood and a node draining a secondary vaccine site (sentinel immunized node) were harvested during the course of vaccination and T-cell responses to the peptides were evaluated using an enzyme-linked immunosorbent spot assay. CD8 T-cell responses were detected in 1 participant ex vivo and in 8 of 9 participants (89%) after in vitro stimulation. All 4 HLA-A2 and HLA-A3 restricted peptides were immunogenic. This includes 2 peptides, folate binding protein (FBP191-199) and Her-2/neu754-762, which had not previously been evaluated in vaccines in humans. Responding T cells required over 200 nM for half maximal reactivity. These data support continued investigation of these peptides as immunogens for patients with ovarian cancer but, owing to low potency, also suggest a need for additional immunomodulation in combination with vaccines to increase the magnitude and to improve the quality of the T-cell responses. PMID- 18391754 TI - Maintenance immunotherapy in recurrent or metastatic squamous cell carcinoma of the head and neck. AB - The purpose of this study was to determine the efficacy and safety of a maintenance immunotherapy regimen administered to patients with recurrent/metastatic squamous cell carcinoma of the head and neck (RMHN) who showed clinical benefit from docetaxel, ifosfamide, and cisplatin chemotherapy (DIP). Every 4 weeks, patients with RMHN received 60 mg/m docetaxel on day 1, and 1200 mg/m ifosfamide and 20 mg/m cisplatin on days 1 to 4. Low-dose subcutaneous interleukin-2 and oral 13-cis-retinoic acid were administered as maintenance immunotherapy to patients who showed a clinical benefit (complete or partial response, disease stability). The primary end point was response; secondary end points were progression-free survival, overall survival, toxicity, and evaluations of lymphocytes, natural killer cells, and serum vascular endothelial growth factor (VEGF). After a median follow-up of 22 months, 263 courses of chemotherapy were administered to the 54 patients. The overall response rate was 59%. Forty-two patients (78%) had a clinical benefit and received 185 courses of maintenance immunotherapy. Median progression-free survival and overall survival were 11.1 and 21.8 months, respectively. Statistically significant, progressive increases in lymphocytes and natural killer cells and a decrease in VEGF were observed in patients treated with maintenance immunotherapy. The toxicity was relatively well tolerated and caused no death. Outpatient administration of DIP, followed by low-dose interleukin-2 and 13-cis-retinoic acid, was generally well tolerated and showed promising activity against RMHN. Longitudinal changes in lymphocytes, natural killer cells, and VEGF might be useful biomarkers for response and survival. PMID- 18391755 TI - Nonmyeloablative chemotherapy followed by T-cell adoptive transfer and dendritic cell-based vaccination results in rejection of established melanoma. AB - We demonstrated previously that dendritic cell (DC)-based vaccines could mediate a specific and long-lasting antitumor immune response during early lymphoid reconstitution after lethal irradiation and bone marrow transplant. The purpose of this current study was to examine the potential therapeutic efficacy of DC based vaccines in combination with sublethal lymphodepletion and T-cell transfer. In an aggressive model of melanoma, treatment with the combination of 200 mg/kg cyclophosphamide (Cy) and 100 mg/kg fludarabine (Flu) led to a lymphopenic state lasting approximately 14 days, but had no effect on the growth of an established M05 melanoma. Addition of ovalbumin (OVA) peptide-pulsed DC-based immunization resulted in a delay in tumor growth but did not enhance overall survival in this model. To improve treatment, adoptively transferred naive T cells were added. After induction of lymphopenia with Cy and Flu, transferred T cells demonstrated an activated memory phenotype including high expression of CD44 and low expression of CD62L. Induction of lymphopenia with Cy and Flu in combination with adoptive transfer of naive T cells and OVA peptide-pulsed DCs immunization led to an enhancement in the number of OVA specific, CD8 T cells that demonstrated specific cytotoxic activity, proliferation, and interferon-gamma production in response to the OVA expressing M05 melanoma. This combination therapy also led to tumor regression and enhanced survival in mice bearing M05 melanoma. PMID- 18391756 TI - Surmounting tumor-induced immune suppression by frequent vaccination or immunization in the absence of B cells. AB - Tumor-induced immune suppression is one of the most difficult obstacles to the success of tumor immunotherapy. Here, we show that established tumors suppress CD8 T cell clonal expansion in vivo, which is normally observed in tumor-free mice upon antigen-specific glycoprotein (gp) 96-chaperone vaccination. Suppression of CD8 T-cell expansion by established tumors is independent of tumor associated expression of the antigen that is recognized by the CD8-T-cell receptor. Vaccination of tumor-bearing mice is associated with increased cellular recruitment to the vaccine site compared with tumor-free mice. However, rejection of established, suppressive tumors required frequent (daily) gp96 vaccination. B cells are known to attenuate T helper cell-1 responses. We found that in B-cell deficient mice, tumor rejection of established tumors can be achieved by a single vaccination. Accordingly, in tumor-free B-cell deficient mice, cognate CD8 cytotoxic T lymphocyte clonal expansion is enhanced in response to gp96-chaperone vaccination. The data have implications for the study of tumor-induced immune suppression and for translation of tumor immunotherapy into the clinical setting. Frequent vaccination with cellular vaccines and concurrent B-cell depletion may greatly enhance the activity of anticancer vaccine therapy in patients. PMID- 18391757 TI - Interleukin-15 favors the expansion of central memory CD8+ T cells in ex vivo generated, antileukemia human cytotoxic T lymphocyte lines. AB - We demonstrated in previous studies that interleukin (IL) -2 supports in vitro cell proliferation of donor-derived cytotoxic T lymphocyte (CTL) lines directed against different types of leukemia blasts. The aim of this study was to compare the capacity of IL-15 with that of IL-2 in supporting the proliferation and cytotoxic activity of antileukemia CTL cultures, and their influence on T-cell memory compartment differentiation. Antileukemia CTL lines were generated using donor-derived dendritic cells pulsed with apoptotic leukemia blasts, in the presence of IL-12 and IL-7, during the primary culture, and expanded through 2 rounds of leukemia-specific stimulation and 1 round of antigen-independent expansion, each supplemented with either IL-2 or IL-15. Both IL-2-supplemented (IL-2-CTLs) and IL-15-supplemented (IL-15-CTLs) lines contained predominant numbers of CD45RA/CCR7 effector memory (TEM) and CD45RA/CCR7 (TEMRA+) T cells. Significantly higher numbers (P<0.05) of CD8-positive central memory T cells (TCM), and higher expansion rate, together with comparable cytotoxic activity, were observed in IL-15-CTLs compared with IL-2-CTLs. Altogether, these results demonstrate that IL-15 enhances recovery of CTL activity, without loss of leukemia-directed specificity, and favors expansion of TCM CD8-positive cells, expected to exhibit long-term survival and differentiation capacity in vivo in the presence of a limited amount of antigen. PMID- 18391758 TI - Efficient adenovector CD40 ligand immunotherapy of canine malignant melanoma. AB - Cutaneous canine melanomas are usually benign in contrast to human malignant melanoma. However, the canine oropharyngeal, uveal, and mucocutaneous neoplasms are aggressive and have metastatic potential. Surgery and to a lesser extent radiotherapy and chemotherapy are widely adopted treatments but are seldom curative in advanced stages. The similarities between human and canine melanoma make spontaneous canine melanoma an excellent disease model for exploring novel therapies. Herein, we report the first 2 adenovector CD40L immunogene (AdCD40L) treatments of aggressive canine malignant melanoma. Case no. 1 was an advanced stage III oral melanoma that was cured from malignant melanoma with 2 intratumor AdCD40L injections before cytoreductive surgery. After treatment, the tumor tissue was infiltrated with T lymphocytes and B lymphocytes suggesting immune activation. This dog survived 401 days after the first round of gene therapy and was free of melanoma at autopsy. Case no. 2 had a conjunctival malignant melanoma with a rapid progression. This case was treated with 6 AdCD40L injections over 60 days. One hundred and twenty days after start of gene therapy and 60 days after the last injection, the tumor had regressed dramatically, and the dog had a minimal tumor mass and no signs of progression or metastasis. Our results indicate that AdCD40L immunogene therapy is beneficial in canine malignant melanoma and could be considered for human malignant melanoma as well. PMID- 18391759 TI - Small cleavable adapters enhance the specific cytotoxicity of a humanized immunotoxin directed against CD64-positive cells. AB - The most potent immunotoxins (ITs) developed to date contain bacterial or plant cytotoxic components. As these are potentially immunogenic in man, human proteins are preferred for the long-term treatment of cancer. We have developed the first humanized IT for the treatment of CD64 malignancies such as acute myeloid leukemia. The bacterially expressed IT is composed of a humanized anti-CD64 single chain fragment [h22(scFv)] genetically fused to the human RNase angiogenin. As angiogenin lacks a dedicated translocation domain responsible for the higher potency of bacterial and plant-derived toxins, we have incorporated a recombinant adapter that contains a synthetic translocation domain flanked by proteolytically cleavable endosomal and cytosolic consensus sites. Although insertion of the adapter increased the cytotoxicity by up to 20-fold, serum stability was markedly reduced. Therefore, we designed a modified adapter variant with the endosomal-cleavable peptide deleted. The IT containing the truncated adapter showed significantly higher cytotoxicity than the adapter-free IT and superior serum stability to facilitate the potential applications in patients. PMID- 18391760 TI - Using dendritic cell maturation and IL-12 producing capacity as markers of function: a cautionary tale. AB - Effective dendritic cell (DC) function depends on sufficient expression of antigen and costimulatory molecules, and secretion of interleukin (IL)-12. We sought to augment DC stimulatory capacity by optimizing DC phenotype and IL-12 production. DCs, obtained by CD14-selection, were matured using 8 different cytokine cocktails, and expression of costimulatory/major histocompatibility complex molecules and IL-12 production at the end of maturation was assessed. DC stimulatory capacity was determined after pulsing with immunogenic adenoviral CD8 peptide epitopes or after transduction with an Ad5f35-null vector. Resultant T cell cultures were analyzed using pentamer and interferon-gamma enzyme-linked immunosorbent spot assays. On the basis of DC expression of maturation markers and IL-12 production, we defined prototype "minimal" [tumor necrosis factor-alpha (TNF-alpha), prostaglandin E2], "standard" (IL-1, IL-6, TNF-alpha, prostaglandin E2), and "optimal" (IL-1, IL-6, TNF-alpha, interferon-alpha, CD40 ligand) DC cocktails. Optimal DCs were functionally superior when pulsed with CD8 peptides, but when transduced with Ad5f35, functioned poorly as antigen-presenting cells. We investigated the mechanisms underlying this discrepancy and suggest that prolonged stimulation with potent cytokines (optimal cocktail) in combination with adenoviral transduction alters the kinetics of DC maturation such that the DCs are functionally exhausted by the traditional 48-hour maturation time point. Shortening the DC maturation period posttransduction restored optimal DC stimulatory capacity. Thus, maturation stimuli and viral transduction affects DC phenotype, IL-12 producing capacity, and kinetics of maturation, and all must be considered before designing protocols to generate the optimal DC for cytotoxic T lymphocyte generation. PMID- 18391761 TI - Mechanisms involved in radiation enhancement of intratumoral dendritic cell therapy. AB - We have previously reported that local tumor irradiation, without inducing cell death, can augment the therapeutic efficacy of intratumoral (IT) dendritic cell (DC) vaccination. This study examined potential mechanisms underlying radiation enhancement of IT DC therapy in this setting. Even though ionizing radiation did not mediate tumor cell killing, bone marrow-derived DCs acquired in vitro tumor antigens from irradiated D5 murine melanoma cells more efficiently than from untreated cells. This radiation-enhanced loading of DCs did not induce DC maturation, but was associated with improved cross-priming of T cells both in vitro and in vivo. Furthermore, in vivo pulsing of DCs with irradiated versus untreated tumor cells resulted in superior presentation of tumor antigens to T cells. In addition, tumor irradiation facilitated homing of IT administered DCs to the draining lymph node, possibly by down-regulating CCL21 expression within the tumor mass. Studies of the tumor microenvironment in irradiated versus untreated tumors did not reveal significant inflammatory changes. Moreover, radiation did not promote accumulation of CD4 or CD8 effector T cells within solid tumors. Our results indicate that, without inducing cytotoxicity, tumor irradiation can enhance the ability of DCs to capture tumor antigens, migrate to the draining lymph node, and present processed antigens to T cells. These findings may prove useful in designing future strategies for human cancer immunotherapy. PMID- 18391762 TI - Migration patterns of nonspecifically activated versus nonactivated nonhuman primate T lymphocytes: preferential homing of activated autologous CD8+ T cells in the rectal mucosa. AB - Adoptive cell transfer may be a successful strategy in anticancer therapy and its therapeutic efficiency depends on the access of transferred cells to the tumor site and their persistence in vivo. Nevertheless, the migration properties of autologous in vitro-activated T cells in primates are largely unknown. Here, we established the long-term tracking of T-cell migration into various compartments of rhesus macaques as a preclinical model for the evaluation of T-cell-based immunotherapy. Peripheral blood mononuclear cells from 3 to 4 rhesus macaques were activated with anti-CD3/anti-CD28 or not, labeled with carboxyfluorescein diacetat succinimidyl ester, and reinjected intravenously into the donor animals. Blood samples, lymph node biopsies, and mucosal biopsies (duodenum, rectum) were collected at various time points and analyzed by flow cytometry for the presence of the reinjected T cells. We demonstrate that nonspecific in vitro activation changes the in vivo migratory behavior of T cells and provokes a preferential migration of CD8 T cells to the rectum. Nonspecifically activated transferred CD4 T cells were found in much lower frequencies at this site and also in other compartments. Thus, our results indicate an imbalanced distribution of autologous CD8 and CD4 T cells in various compartments that is more apparent when T cells are activated before the transfer. The migratory behavior of in vitro-expanded, autologously transferred T cells can, therefore, influence the clinical outcome of adoptive cell transfer. PMID- 18391763 TI - Inducible nitric oxide synthase (iNOS) is not required for IL-2-induced hypotension and vascular leak syndrome in mice. AB - Dose limiting side effects of interleukin-2 (IL-2) include severe hypotension and vascular leak syndrome (VLS). Previous studies have shown that nitric oxide (NO) synthesis is strongly induced after IL-2 treatment of C3H/HeN mice (180,000 IU b.i.d. for 5 d). We employed knockout mice (on C57BL/6 background) to test the role of the inducible NO synthase (iNOS) in mediating IL-2 toxicity. In contrast to C3H/HeN mice, which developed hypotension and VLS after 10 doses of only 180,000 IU IL-2, C57BL/6 mice were far more resistant requiring increased doses of 800,000 IU IL-2 (b.i.d., 5 d) to induce hypotension and VLS. Serum interferon gamma levels were significantly more elevated by IL-2 treatment in C3H/HeN mice than in C57BL/6, correlating with the severity of hypotension and VLS. Urinary excretion of NO metabolites was markedly reduced in iNOS knockout mice (C57BL/6 iNOS) after IL-2 treatment. A surprising finding was that these mice still developed profound hypotension and VLS. Similar findings were observed after administration of a iNOS specific inhibitor, L-N[6]-(1-iminoethyl)lysine (L-NIL). In contrast, a general NOS inhibitor, N-monomethyl-L-arginine, prevented both hypotension and vascular leak. The superoxide dismutase mimetic, M40403, reversed IL-2-induced hypotension but not VLS in knockout mice. Thus, peroxynitrite mediated mechanisms are likely responsible for hypotension, whereas NO-induced changes in vascular permeability result in VLS. The iNOS enzyme is not necessary for pathogenesis of IL-2-induced cardiovascular toxicity. These results imply that other NOS isoforms, such as endothelial NOS, may play a major role in the development of IL-2-induced cardiovascular toxicity. PMID- 18391764 TI - The effect of occult nodal metastases on survival and regional control in patients with head and neck squamous cell carcinoma. AB - OBJECTIVES: To determine factors associated with disease-free survival (DFS) and regional control in clinically node-negative head and neck squamous cell cancer (HNSCC) patients with occult metastasis. STUDY DESIGN: Non-randomized retrospective analysis. MATERIALS AND METHODS: Patients who underwent elective neck dissection (END) from 1985 to 2002 were analyzed. RESULTS: A total of 337 patients underwent END. The majority of patients (67%) had advanced stage disease (T3/T4). Occult metastases were present (pN+) in 168 patients (50%), with extracapsular spread (ECS) present in 72 patients (43%). Five-year DFS for patients with histologically node negative necks was 62% versus 36% for pN+ patients (P < .0001). Postoperative radiation (XRT) did not significantly influence DFS for pN+ patients with less than three nodes involved, but had a significant association with DFS with three or more nodes involved (P < .0001). XRT showed a trend toward improved regional control rates in patients with less than three positive nodes (86% vs. 78%; P = .7579) and patients with three or more positive nodes (62% vs. 50%; P = .0014). When ECS was present, XRT did not affect DFS in patients with less than three nodes (36%), but had a significant effect on DFS in patients with three or more nodes (20% vs. 0%; P = .0075). Regional control rates were not improved with XRT in ECS-positive patients with less than three nodes (62% vs. 75%) or with three or more nodes involved (43% vs. 50%; P = .0678). CONCLUSIONS: There is a high incidence of occult metastases in clinically node-negative patients which adversely affects survival, regardless of the use of adjuvant XRT. Postoperative XRT did not significantly affect regional control or survival rates in patients with <3 positive nodes. When ECS was present, survival was poor regardless of the number of nodes. These data emphasize the prognostic and therapeutic role of END and highlight the need for the development of novel therapeutic regimens to improve disease control and survival in HNSCC patients with nodal metastases. PMID- 18391765 TI - Modified nasal specula and flexible holder for endoscopic nasal surgery. AB - The use of an endoscope in nasal surgery has now extended to surgery on the nasal septum as well as to sinus surgery. We devised the use of the modified nasal specula with Yasargil's flexible holder to resolve the technical problems in using an endoscope for septal surgery. One nasal speculum has two plates for connecting to the Yasargil's flexible holder. This speculum allows the surgeon to use both hands freely and also functions as a guide for stabilizing an endoscope. Another nasal speculum has two long adjustable wings. This speculum can widen the working space between the cartilage or bone of the nasal septum and detached mucosa. Our modified nasal specula are readily applicable to resolve the problems such as unstable endoscope and narrow working space in septal surgery under endoscopic view. PMID- 18391766 TI - Day float: an alternative to the night float coverage system for residency programs. AB - OBJECTIVES: The Accreditation Council for Graduate Medical Education (ACGME) has mandated an 80-hour work week that has resulted in changes to many residency programs. In otolaryngology, most programs have switched to either home call or night float systems. Our department covers all of the maxillofacial trauma and backup airway calls, which has made it difficult to employ a home call system. Instead of a night float coverage system, our program implemented a day float coverage system that allows the residents to participate in a 24-hour call period. After call and sign-out, the residents go home; however, their clinical duties are covered by the day float resident. STUDY DESIGN: A brief review of the literature pertaining to call coverage systems followed by a description of our day float system. Residents who have participated in either night float, day float, or both systems were then surveyed regarding their experiences or perceptions of both systems. METHODS: A nine-question survey was handed out to our otolaryngology residents and their responses were recorded. RESULTS: The averaged responses strongly favored the day float over the night float coverage system regardless of the level of training and the systems in which the residents have participated. CONCLUSIONS: The day float coverage system is favored by residents in our program. It allows for a more attending-like 24-hour period of call, continuity of care, attendance at educational activities, and more time with family. In addition, it eliminates a prolonged period devoid of clinical activities. PMID- 18391767 TI - Image-guidance for endoscopic sinus surgery. AB - OBJECTIVES/HYPOTHESIS: Image-guided surgery (IGS) is a critical tool. However, its clinical validity remains controversial. A review of indications and a consensus based on literature are presented. METHODS: Literature review. RESULTS: Traditional indications remain valid. Novel applications have emerged, and the reliability of IGS has been consistently endorsed. Although evidence is based on scattered reports and expert opinions, the use of IGS in approved instances is regarded as the state of the art. CONCLUSIONS: IGS is reliable, and a scientific validation of outcomes would entail unethical strategies. An expanding spectrum of indications is anticipated, although IGS does not replace proper training. PMID- 18391768 TI - Gene expression profiling of nasal polyps associated with chronic sinusitis and aspirin-sensitive asthma. AB - OBJECTIVE: To identify genes whose expression is most characteristic of chronic rhinosinusitis and aspirin-sensitive asthma through genome-wide transcriptional profiling of nasal polyp tissue. STUDY DESIGN: Prospective, controlled study conducted at a tertiary care institution. METHODS: Thirty genome-wide expression microarrays were used to compare nasal polyp tissue from patients with chronic rhinosinusitis alone (CRS, n = 10) or chronic rhinosinusitis and a history of aspirin-sensitive asthma (ASA, n = 10) to normal sinonasal mucosa from patients who underwent surgery for non-sinus related conditions (controls, n = 10). Genes found to be most characteristic of each polyp phenotype, as determined from bioinformatic analyses, were validated using real-time quantitative polymerase chain reaction (RT-PCR) and immunohistochemistry in different patient sets. RESULTS: The transcriptional signature of the control mucosa was distinctly different from that of either polyp phenotype. Genes most characteristic of the CRS phenotype included two upregulated genes--met proto-oncogene (MET) and protein phosphatase 1 regulatory subunit 9B (PPP1R9B)-and two downregulated genes -prolactin-induced protein (PIP) and zinc alpha2-glycoprotein (AZGP1). The gene most characteristic of the ASA phenotype was periostin (POSTN), which was upregulated relative to controls. Differences between the CRS and ASA phenotypes were associated with alterations in the 6p22, 22q13, and 1q23 chromosomal regions. CONCLUSIONS: Nasal polyps appear to have characteristic transcriptional signatures compared to normal sinonasal mucosa. The five genes identified in this study likely play roles in the pathogenesis of polyps associated with CRS and ASA, and are therefore attractive targets for novel medical therapies for these common debilitating diseases. PMID- 18391769 TI - Endoscopic removal of juvenile nasopharyngeal angiofibromas: a video presentation. AB - OBJECTIVES/HYPOTHESIS: Juvenile nasal angiogibroma (JNA) is a relatively rare, benign neoplasm generally seen in prepubertal and adolescent males. In the past, surgical management included lateral rhinotomy, transpalatal and transmaxillary routes, and midface degloving approaches. Endoscopic approaches have become the treatment of choice, especially for tumors limited by size and location criteria. Our presentation demonstrates an endoscopic approach to excision of JNAs that are limited to the nasal cavity, nasopharynx, and paranasal sinuses. Through the use of a brief video presentation, participants will be able to visualize the endoscopic technique. STUDY DESIGN AND METHODS: A retrospective review of JNA patients treated at our institution identified 3 patients who underwent endoscopic removal. All patients underwent preoperative embolization followed by endoscopic resection. RESULTS: A video demonstration is presented of a patient with a 3.5 cm JNA who underwent successful endoscopic, endonasal excision. After medialization of the middle turbinate and anterior ethmoidectomy, the tumor could be mobilized using a Cottle elevator and microdebrider. The origin and attachment of the tumor were visualized and detached from the sphenopalatine foramen. The tumor was resected en bloc with minimal bleeding. There were no intraoperative complications, and postoperative bleeding was negligible. To date, there has been no evidence of recurrence of tumor in these individuals. CONCLUSIONS: Endoscopic removal is the approach of choice for small to medium, limited-extent JNAs. PMID- 18391778 TI - Ontogeny of erythropoiesis. AB - PURPOSE OF REVIEW: The present study review examines the current understanding of the ontogeny of erythropoiesis with a focus on the emergence of the embryonic (primitive) erythroid lineage and on the similarities and differences between the primitive and the fetal/adult (definitive) forms of erythroid cell maturation. RECENT FINDINGS: Primitive erythroid precursors in the mouse embryo and cultured in vitro from human embryonic stem cells undergo 'maturational' globin switching as they differentiate terminally. The appearance of a transient population of primitive 'pyrenocytes' (extruded nuclei) in the fetal bloodstream indicates that primitive erythroblasts enucleate by nuclear extrusion. In-vitro differentiation of human embryonic stem cells recapitulates hematopoietic ontogeny reminiscent of the murine yolk sac, including overlapping waves of hemangioblast, primitive, erythroid, and definitive erythroid progenitors. Definitive erythroid potential in zebrafish embryos, like that in mice, initially arises prior to, and independent of, hematopoietic stem cell emergence in the region of the aorta. Maturation of definitive erythroid cells within macrophage islands promotes erythroblast-erythroblast and erythroblast-stromal interactions that regulate red cell output. SUMMARY: The study of embryonic development in several different model systems, as well as in cultured human embryonic stem cells, continues to provide important insights into the ontogeny of erythropoiesis. Contrasting the similarities and differences between primitive and definitive erythropoiesis will lead to an improved understanding of erythroblast maturation and the terminal steps of erythroid differentiation. PMID- 18391780 TI - Iron regulation and erythropoiesis. AB - PURPOSE OF REVIEW: The peptide hormone hepcidin regulates iron metabolism in response to erythropoietic demand, iron stores and inflammation. Major advances have been made in understanding the regulation of hepcidin production, and consequently the availability of iron for erythropoiesis. RECENT FINDINGS: It is becoming clear that the bone morphogenetic protein (BMP) pathway plays a major role in setting the baseline hepcidin level and, with the assistance of BMP2/4 and hemochromatosis-related proteins hemojuvelin, HFE and transferrin receptor 2, also regulates hepcidin expression in response to iron. Regulation of hepcidin in anemias has now been linked to increased erythropoietic activity and is likely mediated by factor(s) secreted by erythroid precursors. GDF-15 was identified as a candidate for one of the erythroid factors suppressing hepcidin. Tissue hypoxia may also directly contribute to hepcidin suppression in anemias. Regulation of hepcidin by inflammation may include multiple cytokines and the Toll-like receptors pathways. Although it has not yet been shown that increased hepcidin is indispensible for the development of anemia of inflammation, transgenic overexpression of hepcidin was sufficient to replicate its key features. SUMMARY: Regulation of hepcidin and iron availability for erythropoiesis has revealed unexpected pathways and much complexity. The renaissance of the study of iron regulation continues to reward researchers with interesting biology and potential therapeutic targets. PMID- 18391779 TI - Aplastic anemia. AB - PURPOSE OF REVIEW: Most acquired aplastic anemia is the result of immune-mediated destruction of hematopoietic stem cells causing pancytopenia and an empty bone marrow, which can be successfully treated with either immunosuppressive therapy or hematopoietic stem-cell transplantation. RECENT FINDINGS: In aplastic anemia, oligoclonally expanded cytotoxic T cells induce apoptosis of hematopoietic progenitors. T-bet, a transcription factor that binds to the interferon-gamma promoter region, is upregulated in aplastic anemia T cells. Regulatory T cells are significantly reduced in patients' peripheral blood and in an aplastic anemia murine model, infusion of regulatory T cells ameliorates disease progression. In a minority of cases, loss-of-function mutations in telomerase complex genes may underlie disease development. Long-term survival, once strongly linked to response to immunosuppressive therapy, can now be achieved even among nonresponders due to significant advances in supportive care and better salvage treatments. SUMMARY: Evidence has accumulated in the recent years further corroborating an immune-mediated process underlying aplastic anemia pathogenesis. Hematopoietic stem-cell transplantation from a matched sibling donor is preferred for children and young adults with severe aplastic anemia, and immunosuppressive therapy is employed when hematopoietic stem-cell transplantation is not feasible due to age, lack of a histocompatible sibling, co-morbidities, or by patient choice. PMID- 18391781 TI - Long-range regulation of alpha globin gene expression during erythropoiesis. AB - PURPOSE OF REVIEW: The analysis of globin gene expression during erythropoiesis has established many principles underlying normal mammalian gene expression. New aspects of gene regulation have been revealed by natural mutations that downregulate globin gene expression and cause thalassemia. Deletions involving sequences upstream of the alpha and beta clusters suggested that the globin genes might be controlled by remote regulatory elements. This was demonstrated experimentally and suggested that many mammalian genes may be controlled in a similar manner. RECENT FINDINGS: Completion of the Human Genome Project and the associated encyclopaedia of DNA elements (ENCODE) project confirmed that human gene expression is commonly controlled by long-range, cis-acting elements. The development of chromatin immunoprecipitation has allowed us to identify binding of transcription factors and chromatin modifications at the key cis-acting sequences in vivo. In addition, chromosome conformation capture has enabled us to address the topological models proposed to mediate long-range interactions. Together, these methods have given us some insight into how long-range elements may influence gene expression and how this process may be subverted in thalassemia. SUMMARY: The review asks how remote elements regulate alpha globin expression and how natural mutations interfere with this mechanism to cause alpha thalassemia. We also speculate as to why long-range control of gene expression may have evolved in higher organisms. PMID- 18391782 TI - Red cell membrane transport abnormalities. AB - PURPOSE OF REVIEW: The present review describes the red cell transport abnormalities of proteins of the band 3 macrocomplex. The macrocomplex is involved in red cell gas exchange and recent findings have furthered our understanding of this process. RECENT FINDINGS: Study of a novel band 3 hereditary spherocytosis variant suggests that expression of mutant band 3 protein can be rescued by wild-type band 3. Other studies show that some mutant band 3 protein can mediate a cation conductance. Recent work suggests both Rh associated glycoprotein and aquaporin act as gas channels confirming the integrated function of the macrocomplex and the importance of its role in red cell gas transport. SUMMARY: The most recent studies on band 3-induced hereditary spherocytosis are reviewed and an explanation for the mild phenotype of heterozygous hereditary spherocytosis is discussed. A number of red cell conditions (hereditary stomatocytosis, south-east Asian ovalocytosis, distal renal tubular acidosis, Rhnull), associated with both stomatocytosis and a cation leak, are described. The evidence that Rh-associated glycoprotein forms a gas channel that transports CO2 and/or NH3 is reviewed and discussed, together with recent studies that show that aquaporin 1 transports both CO2 and O2. PMID- 18391783 TI - Regulation of phospholipid asymmetry in the erythrocyte membrane. AB - PURPOSE OF REVIEW: Alterations in the transbilayer distribution of phospholipids in the erythrocyte membrane have significant physiologic consequences. Understanding the cause of these perturbations and the molecular mechanisms by which they are regulated is essential for ameliorating some of the consequences of erythrocyte membrane abnormalities. This review summarizes recent data that provide a clearer description of the molecular events involved in these processes. RECENT FINDINGS: Proteomic and molecular evidence supports the presence of a small number of lipid transporters that control the distribution of phospholipids across the erythrocyte membrane. These include members of the ATP binding cassette and P-type ATPase transporter families. Recent data indicate that these proteins work in synergy and may require additional protein partners. Growing evidence illustrates the critical role these transporters play in the maintenance of lipid asymmetry and the central role that oxidative damage plays in the membrane perturbing effects of some cardiovascular diseases. SUMMARY: Recent publications have added further clarity to the processes that control phospholipid asymmetry in erythrocytes and the mechanisms by which diseases affect lipid distribution. Transmembrane transporters regulate the organization of phospholipids across the bilayer, and oxidative damage induced by disease states may be a common perturbant of phospholipid asymmetry. PMID- 18391784 TI - Vascular biology is moving forward at a very rapid pace. Editorial comment. PMID- 18391785 TI - Extracellular matrix mediates a molecular balance between vascular morphogenesis and regression. AB - PURPOSE OF REVIEW: We discuss very recent studies that address the critical role of extracellular matrix in controlling the balance between vascular morphogenesis and regression. Much of this work suggests that a balance mechanism exists for controlling the extent of tissue vascularization involving downstream signaling events regulating endothelial cell behaviors in relation to their interactions with extracellular matrix molecules. RECENT FINDINGS: Endothelial gene expression changes and signaling lead to events that not only stimulate vascular morphogenesis but also suppress mechanisms mediated through pro-regression factors such as Rho kinase. At the same time, vascular networks are susceptible to regression mediated by factors such as matrix metalloproteinase-1, matrix metalloproteinase-10, thrombospondin-1, extracellular matrix matricryptic fragments and angiopoietin-2. Pericyte recruitment to such vascular tubes can prevent regression events by delivering molecules such as tissue inhibitor of metalloproteinase-3 and angiopoietin-1 that promote vascular stabilization by decreasing tube susceptibility to these regression stimuli. SUMMARY: Extracellular matrix-derived signals lead to critical morphologic changes mediated through cytoskeletal rearrangements that control the shape, function and signaling events in endothelial cell-lined vessels regulating tube formation, remodeling, stabilization and regression. These signals control both vascular morphogenic and regression events, and thus a molecular balance exists to control the extent and function of vascular tube networks within tissues. PMID- 18391786 TI - Endothelial-stromal interactions in angiogenesis. AB - PURPOSE OF REVIEW: Angiogenesis often occurs in the context of a wound or tumor stroma. This review will focus on the recent findings on the interactions between angiogenic endothelial cells and the other components of the stroma - fibroblasts, pericytes and extracellular matrix. RECENT FINDINGS: Large-scale gene expression arrays have provided a remarkable insight into the diversity of fibroblasts in different tissues and under different conditions. These somewhat neglected cells are now understood to play a critical role in tumor growth, regulating not only the phenotype of the tumor cells but also the angiogenic response that supports them. These advances are leading to an understanding of the soil and seed hypothesis at the molecular level. In addition, there is a new focus on the role of pericytes in regulating angiogenesis and their potential as targets for tumor therapy. SUMMARY: Initiation of new blood vessel formation requires metalloproteinase induction leading to the degradation of the basement membrane, sprouting of endothelial cells and regulation of pericyte attachment. Fibroblasts and their activated counterpart, the myofibroblast, play a large role in synchronizing these events through the expression of numerous extracellular matrix molecules, growth factors and morphogens, including fibroblast growth factors and transforming growth factor beta. PMID- 18391787 TI - Pleiotrophin, a multifunctional angiogenic factor: mechanisms and pathways in normal and pathological angiogenesis. AB - PURPOSE OF REVIEW: This study seeks to integrate recent studies that identify new critical mechanisms through which the 136 amino acid secreted heparin-binding cytokine pleiotrophin (PTN, Ptn) stimulates both normal and pathological angiogenesis. RECENT FINDINGS: Pleiotrophin is directly angiogenic; it initiates an angiogenic switch in different cancer models in vivo. It acts as an angiogenic factor through multiple mechanisms that include a unique signaling pathway that activates newly identified downstream tyrosine kinases through a unique mechanism, an interaction with endothelial cells to initiate proliferation, migration, and tube formation, the regulation of basic fibroblast growth factor and vascular endothelial growth factor signaling, the remodeling of the stromal microenvironment, and induction of transdifferentiation of monocytes into endothelial cells. Recently also, domains of PTN that stimulate angiogenesis and peptides that function to inhibit PTN signaling have been identified. SUMMARY: Recent studies have identified new mechanisms dependent on activation of the PTN signaling pathway that regulate angiogenesis and new targets to use PTN to both stimulate angiogenesis and block its activity to control pathological angiogenesis. PMID- 18391789 TI - Deciphering arterial identity through gene expression, genetics, and chemical biology. AB - PURPOSE OF REVIEW: The present review presents a current view of vascular development, with a focus on the factors contributing to the establishment of arterial-venous identity and the potential of chemical biology for providing new insights into this field. RECENT FINDINGS: Genetics and gene expression studies have begun to define the complex network of molecular pathways that govern the formation of the embryonic vasculature, but these approaches have limited ability to spatially and temporally manipulate gene expression and function. Recently, the power of chemical biology, combined with model systems like zebrafish, has enabled discovery of additional contributors to vascular development and has provided a means of manipulating gene function with enhanced spatial and temporal control. SUMMARY: The molecular pathways directing arterial-venous specification during embryogenesis are relevant for understanding the causes of human arteriovenous malformations, tumor angiogenesis, and diabetic retinopathy. Through the complementary strengths of genetics and chemical biology, it is hoped that novel therapeutic approaches for these conditions will emerge. PMID- 18391790 TI - Rho signaling and mechanical control of vascular development. AB - PURPOSE OF REVIEW: To discuss how mechanical cues and Rho signaling contribute to control of vascular development and hematopoiesis. RECENT FINDINGS: Rho guanine trinucleotide phosphatases are ubiquitious regulators of cytoskeletal structure and tension generation. Recent work shows that Rho-dependent mechanical interactions between cells and extracellular matrix regulate cell fate switching in capillary endothelial cells and megakaryocytes in vitro, as well as angiogenesis, vascular permeability, leukocyte migration and platelet formation in vivo. Signaling pathways that link integrins and tension-dependent changes in cytoskeletal structure to Rho have also begun to be delineated. SUMMARY: Mechanical force generation by cells and simultaneous sensing of these physical forces play critical roles in vascular development by estimating whether individual cells will grow, differentiate, move or undergo apoptosis in the local tissue microenvironment. Future work in the vascular field therefore needs to incorporate physical control mechanisms into existing biochemical concepts of cell and tissue regulation. PMID- 18391788 TI - Fibroblast growth factor regulation of neovascularization. AB - PURPOSE OF REVIEW: Fibroblast growth factors are potent angiogenic inducers; however, their precise roles in angiogenesis have not been well understood. In this review, we will focus on specific roles played by fibroblast growth factors in neovascularization. RECENT FINDINGS: Although fibroblast growth factors promote a strong angiogenic response, it has been suggested that FGF-induced angiogenesis requires activation of the vascular endothelial growth factor system. Recent findings have endorsed the view of indirect contribution of fibroblast growth factor signaling to vascular development. A study using embryoid bodies demonstrated a nonimmediate role played by fibrobalst growth factor receptor 1 in vasculogenesis as vascular endothelial growth factor supplementation was sufficient to promote vascular development in Fgfr1-/- embryoid bodies. Moreover, another line of evidence indicated that myocardial fibroblast growth factor signaling is essential for mouse coronary development. The key role of fibroblast growth factor signaling in this process is Hedgehog activation, which induces vascular endothelial growth factor expression and formation of the coronary vasculature. In addition to vascular endothelial growth factor interaction, fibroblast growth factors can control neovascularization by influencing other growth factors and chemokines such as platelet-derived growth factor, hepatocyte growth factor and monocyte chemoattractant protein-1, contributing to development of mature vessels and collateral arteries. SUMMARY: Although fibroblast growth factors are potent angiogenic factors, they may indirectly control neovascularization in concert with other growth factors. Thus, the unique role played by fibroblast growth factors might be organization of various angiogenic pathways and coordination of cell-cell interactions in this process. PMID- 18391791 TI - Integrins team up with tyrosine kinase receptors and plexins to control angiogenesis. AB - PURPOSE OF REVIEW: Understanding the role of integrins in the formation of vascular bed is important for designing new therapeutic approaches to ameliorate or inhibit pathological vascularization. Besides regulating cell adhesion and migration, integrins dynamically participate in a network with soluble molecules and their receptors. This study summarizes recent progress in the understanding of the reciprocal interactions between integrins, tyrosine kinase, and semaphorin receptors. RECENT FINDINGS: During angiogenic remodeling, endothelial cells that line blood vessel walls dynamically modify their integrin-mediated adhesive contacts with the surrounding extracellular matrix. During angiogenesis, opposing autocrine and paracrine loops of growth factors and semaphorins regulate endothelial integrin activation and function through tyrosine kinase receptors and the neuropilin/plexins system. Moreover, proangiogenic and antiangiogenic factors can directly bind integrins and regulate endothelial cell behavior. Studies describing these intense research areas are discussed. SUMMARY: Alteration in the balance between the angiogenic growth factors and semaphorins results in an impairment of integrin functions and could account for cardiovascular malformation and structural and functional abnormalities of the tumor vasculature. PMID- 18391792 TI - Vascular permeability in cardiovascular disease and cancer. AB - PURPOSE OF REVIEW: Regulation of endothelial barrier function is critical for vascular homeostasis, as dynamic and local control of vascular permeability permits macromolecular transport, immune surveillance, and deposition of a fibrin barrier to contain infection at sites of inflammation. Many of the signaling pathways promoting useful vascular permeability, however, are also triggered during disease, resulting in prolonged or uncontrolled vascular leak. Hyperpermeability triggered by inflammation or ischemia in the heart, brain, or lung promotes edema, exacerbates disease progression, and impairs recovery. During cancer, solid tumors release factors that promote the growth of leaky blood vessels which contribute to metastatic spread and limit targeted delivery of anticancer agents. RECENT FINDINGS: Although the molecular mechanisms governing vascular leak have been studied intensely over the last few decades, recent advances have identified new therapeutic targets that have begun to show preclinical and clinical promise. These approaches have been recently applied with success to an increasing number of disease models. SUMMARY: Designing new therapies to limit vascular leak during the progression of disease requires a more complete understanding of the molecular mechanisms governing the endothelial barrier function. This knowledge will benefit the treatment of a growing number of diseases from cardiovascular disease to cancer. PMID- 18391793 TI - Smooth muscle apoptosis and vascular remodeling. AB - PURPOSE OF REVIEW: The present review is to summarize recent advances in molecular mechanisms that regulate vascular smooth muscle cell apoptosis during vascular remodeling. In normal blood vessels apoptosis counteracts cell division, whereas apoptosis is especially crucial for regulating vascular remodeling during cardiovascular diseases. RECENT FINDINGS: Recent results have expanded our knowledge regarding the signaling pathways and molecules that regulate vascular smooth muscle cell death in postnatal vascular remodeling. Compelling data from genetic disorders associated with vascular smooth muscle cell loss (e.g., Hutchinson-Gilford progeria syndrome) and experimental studies suggest that changes in hemodynamic and mechanical forces are major modulators for vascular smooth muscle cell apoptosis. Furthermore, understanding the therapeutic effects of antihypertensive drugs related to apoptosis may identify pathways that can improve outcomes independent of the blood pressure fall. SUMMARY: Regulation of vascular smooth muscle cell apoptosis is a potential target to modify pathological vascular remodeling and new drugs development. PMID- 18391794 TI - Current world literature. Vascular biology. PMID- 18391795 TI - A focus on pediatric gastroenterology nursing. PMID- 18391796 TI - Growth and early onset inflammatory bowel disease. AB - In pediatrics, growth is considered one of the most important markers of overall well-being. This study looked at growth in children diagnosed with inflammatory bowel disease before they were 5 years old from a single center. The Children's Inflammatory Bowel Disease Center at Mount Sinai maintains a database of 1,150 children followed at the center. Ninety-three children were included in this study, 58% boys and 42% girls. The average age at diagnosis was 3.2 years. Sixty two percent had ulcerative colitis and 38% had Crohn disease. Height was recorded at initial presentation and at the most recent visit to the center; from this, a height percentile and z score were calculated. A target adult height was calculated for each child on the basis of mid-parental height. This target height was compared to the actual height the children achieved or the percentile they were growing along. Ten percent of children in the study presented with growth failure. For children with early onset ulcerative colitis, 58% achieved or exceeded their projected height percentile. For children with early onset Crohn disease, 38% achieved or exceeded their projected height percentile. Fifty-nine percent of the entire group either maintained their presentation percentile or increased their height percentiles over time, with an increase in z score ranging from 0.093 to 4.137. PMID- 18391798 TI - Taking care of the little things: preparation of the pediatric endoscopy patient. AB - The practice of pediatric endoscopy offers unique challenges in both the physiological and emotional well-being of children and their families. Successful procedures result in part from providing thorough preprocedure education and preparation. Staff preparing these patients must be knowledgeable about the developmental and physiological needs of each age group of patients entrusted into their care. Awareness and understanding of the developmental approaches to their care is essential to provide positive outcomes for children and their families. This article discusses preparation of the child undergoing endoscopic procedures and interventions to assist in positive outcomes. PMID- 18391799 TI - Pediatric nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease related to excessive accumulation of hepatic fat, and represents a spectrum of liver disease ranging from fat accumulation alone (steatosis) to the more significant histologic finding of steatohepatitis. Nonalcoholic steatohepatitis is a progressive liver disease associated with increased risk of liver cirrhosis and cancer. NAFLD is becoming increasingly prevalent in the pediatric population in direct correlation with the emergence of childhood obesity as a significant pediatric health problem. The exact pathophysiology of NAFLD remains unclear, although the interplay of insulin resistance, oxidative stress, and release of proinflammatory cytokines are implicated in the process. The diagnostic workup and treatment for NAFLD and nonalcoholic steatohepatitis remains controversial. This review discusses current concepts regarding the natural history, pathophysiology, and management of pediatric patients with NAFLD. PMID- 18391800 TI - When the going gets tough: pediatric constipation and encopresis. AB - Constipation and encopresis are two common conditions seen in the pediatric gastroenterology setting. Organic causes cannot be excluded although they are rarely diagnosed in infants and children with defecation disorders. To successfully treat these disorders, a combination of family education, disimpaction and maintenance medications, a well-balanced diet, and behavior management is essential. PMID- 18391801 TI - Pediatric celiac disease. AB - Celiac disease is a disease of the small intestine caused by an immune response to ingested gluten. This response results in characteristic damage to the villi, resulting in malabsorption. It is far more prevalent and its presentation can be far more subtle than once recognized. The prevalence of celiac disease in children in the general U.S. population is estimated to be 1 in 100. Toddlers and young children classically present with failure to thrive, diarrhea, and abdominal distension sometime after the introduction of gluten in the diet, but more often they present with subtle gastrointestinal symptoms such as constipation. Some children may be asymptomatic. Symptomatic patients and high risk individuals should have serologic testing and testing of their total immunoglobulin A. Seropositive patients should undergo upper endoscopy and small bowel biopsy of the distal duodenum. Treatment is lifelong adherence to a gluten free diet. Families must meet with a dietician and should be strongly encouraged to participate in celiac support groups. The gastroenterology nurse is in an unparalleled position to assist parents of children newly diagnosed with celiac disease to identify the necessary lifestyle changes to make their child's life gluten free. PMID- 18391803 TI - The power of less than 1 hour. PMID- 18391804 TI - E-mails from college. Managing inflammatory bowel disease in college. PMID- 18391805 TI - The importance of the literature search. PMID- 18391806 TI - Evolution of walking ability after soft tissue surgery in cerebral palsy patients: what can we expect? AB - Eleven patients with spastic cerebral palsy were evaluated preoperatively, and 3 and 9 months postoperatively after soft tissue surgery. Evaluation included clinical examination, the Functional Mobility Scale questionnaire, and instrumented gait and center of mass trajectory analysis. A decrease in time distance parameters after 3 months was followed by progress in all parameters at 9 months postoperatively. Push-off range of ankle motion decreased after surgery and was not restored to preoperative level until 9 months later. The center of mass vertical displacement improved significantly. The Functional Mobility Scale showed gait improvement. Despite the normalization of range of motion after surgery, there is an obvious period of functional gait deterioration in the early postoperative period and the push-off range of motion at the ankle did not recover to preoperative level until 9 months later. PMID- 18391807 TI - Hip arthrography in the assessment of children with developmental dysplasia of the hip and Perthes' disease. AB - In our unit, children with developmental dysplasia of the hip or Perthes' disease, for whom an operation is considered, undergo examination under anaesthetic and a hip arthrogram. This prospective study assessed whether the arthrogram modified treatment and analysed the reliability of its interpretation. All children undergoing a hip arthrogram for developmental dysplasia of the hip and Perthes' disease over a 12-month period were included. Treatment plans before and after the arthrogram were compared for each of the three children's consultants. The preoperative and arthrographic appearances were blindly reviewed to monitor reproducibility. Twenty-one patients with developmental dysplasia of the hip and 19 with Perthes' disease were included. The treatment plan was modified in 12 of the 21 (57.1%) patients with developmental dysplasia of the hip as a consequence of the arthrogram and six of the 19 patients (31.6%) with Perthes' disease. Intraobserver consultant agreement was high but interobserver agreement was only moderate. Static and dynamic arthrography helps decision making in patients with developmental dysplasia of the hip and Perthes' disease. PMID- 18391808 TI - Perthes' or Perthes' resembling disease in 13-month-old boy. AB - Onset of Perthes' disease is reported frequently from the age of 2 years. Latest publications showed cases with onset of this disease in infancy at ages of 17 and 18 months. We report the case of a 13-month-old boy, who presented with left sided limping. Radiological examination showed reduced height and fragmentation of the femoral head. Magnetic resonance imaging showed the typical signs of an avascular necrosis. Follow-up was done after 3, 7 and 15 months. Plain radiography showed the femoral head in a state of reparation. This is the youngest documented case of Legg-Calve-Perthes' disease and is discussed under consideration of the current literature. PMID- 18391809 TI - Eosinophilic granuloma of the ischium in an infant. AB - This report presents an infant with a solitary eosinophilic granuloma (EG) in an unusual localization at an unusual age treated solely by vinblastine. A solitary EG of the right ischium was detected in a 17-month-old infant as confirmed by a computerized tomography-guided biopsy. He was treated with vinblastine, which was given 6 mg/m per week for a total of six doses. A complete cure was obtained as confirmed by a disease-free follow-up period of 5 years. Chemotherapy is an effective treatment option for a solitary EG in young children considering the risk of dissemination. Moreover, if the disease involves an area where surgery is considered difficult and morbid, or if it involves soft tissues, single-drug chemotherapy can be considered for treatment. PMID- 18391810 TI - Knee synovectomy in children with juvenile idiopathic arthritis. AB - The purpose of the study was to assess the outcomes of knee synovectomies in children with juvenile idiopathic arthritis. Thirty-one arthroscopic synovectomies were performed in 19 children (six oligoarthritis, 20 polyarthritis, five psoriatic arthritis). The percentage of recurrence in the group with oligoarthritis was 67%, in the group with polyarthritis was 95%, whereas all psoriatic arthritis recurred. The overall mean survival (i.e. free from recurrence) was 1.05 years (95% confidence interval, 0.74-1.35). Mean survival time was 1.69, 0.80 and 1.30 years, respectively, for oligoarthritis, polyarthritis and psoriatic arthritis. After synovectomy we observed two complications: thrombophlebitis of the omolateral superior femoral vein and septic arthritis. In conclusion, the mainstay of therapy for juvenile idiopathic arthritis remains medical treatment and intensive physiotherapy. The aim of arthroscopic synovectomy is to allow to make the most of nonsurgical therapy. It revealed more accurate and less invasive results than open synovectomy, maintained the range of motion of the joint, allowed early mobilization and required shorter hospitalization. Best results were observed in the group of oligoarthritis. PMID- 18391811 TI - Solitary intra-articular lipoma locking the knee in a young boy. AB - Intra-articular lipomas have been described in adult patients: only in five of them was it responsible for acute pain and locking of the knee. We report here the unique case of an 8-year-old boy with a painful locked knee caused by a solitary intra-articular lipoma in the anteromedial compartment of the knee. PMID- 18391812 TI - Early results of the Ponseti method using the Steenbek foot abduction brace: a prospective study of 95 feet. AB - The purpose of this study is to evaluate the early results of the Ponseti method and the effectiveness of the Steenbek foot abduction brace. A total of 74 patients with 110 idiopathic clubfeet were included in this prospective study. The feet were evaluated according to the Dimeglio-Bensahel classification, the Catteral-Pirani classification and the functional classification of the Hospital for Joint Diseases. Ninety-eight feet (89%) had a good result after the casting period. All the feet evaluated after the period of full-time bracing and during the period of part-time bracing showed a good correction. The Ponseti method using the Steenbek foot abduction brace is effective in correcting idiopathic clubfeet. PMID- 18391813 TI - Seat load characteristics in children with neuromuscular and syndrome-related scoliosis: effects of pathology and treatment. AB - The amount of correction from spine deformity surgery that is required to avoid asymmetric and unbalanced sitting is not known, and methods sensitive enough to document moderate changes in pressure distribution over time are few, as the commonly used methods are qualitative or semiquantitative. The aim was to analyse seating pressure distribution with a pressure sensor mat system in patients with nonidiopathic scoliosis and to compare the pressure distribution with that in able-bodied controls. The aim was also to apply the method in a surgically treated scoliosis group before and after the spine fusion. Peak pressure, contact area of the entire and the maximum pressure surface, and three measures of asymmetry were calculated. Patients had higher peak pressure and more asymmetric loading than controls. Significant difference was indicated in contact area. After the spine fusion, all asymmetry indices improved. The method fulfilled clinical needs to detect the differences from normal controls as well as the surgery-dependent changes. PMID- 18391814 TI - The growth rate of the humerus: long-term follow-up of treatment of solitary bone cyst of the proximal humerus using cannulated screws: a case report. AB - Although the growth of the proximal epiphysis of the humerus is thought to contribute 80% of the total increase in humeral length, few articles have provided evidence for this. A 9-year-old boy with a pathological fracture at the humeral neck, owing to a solitary bone cyst, was treated by the decompression method using cannulated screws for 7.5 years. During this period, we measured the longitudinal humeral bone growth. The value obtained for the longitudinal growth contribution of the proximal humerus was 88% in this particular case. PMID- 18391815 TI - Focal fibrocartilaginous dysplasia in the humerus. AB - Focal fibrocartilaginous dysplasia is an uncommon, benign bone lesion that causes deformity of the long bones in young children. It has most commonly been encountered in the proximal tibia, and very rarely in the long bones of the upper limb, that is, the proximal humerus, distal radius, ulna, and proximal phalanx. Only one case of focal fibrocartilaginous dysplasia of the proximal humerus has been reported previously. The present study reports two such additional cases that were diagnosed in late childhood. The clinical presentation and radiographic findings are described with an emphasis on the natural evolution of the disease. Limb-length discrepancy is anticipated in these children in the long-term follow up and, therefore, surgical intervention should be considered in the treatment. PMID- 18391816 TI - Humeral lengthening and deformity correction in Ollier's disease: distraction osteogenesis with a multiaxial correction frame. AB - A case of Ollier's disease with deformity and shortening of the humerus is presented. Lengthening of 9 cm and deformity correction of 50 degrees were accomplished with excellent functional and cosmetic results. Unique features of this case were the use of a multiaxial correction monolateral frame and the formation of normal bone within the region of diseased Ollier's bone. PMID- 18391817 TI - Disciplinary action: a tedious task. PMID- 18391819 TI - Research study: professional values and retention. PMID- 18391820 TI - Take a cue from the NDNQI. PMID- 18391821 TI - Legalities of emergency pain management. PMID- 18391822 TI - Incorporating "pull" strategies into clinical education. PMID- 18391824 TI - Shared governance: a journey with continual mile markers. PMID- 18391825 TI - Time = MUSCLE the case for STEMI care improvements. PMID- 18391827 TI - The Magnetic pull: a staff report. PMID- 18391828 TI - Deploying med reconciliation. PMID- 18391829 TI - Eleven strategies for building a personal power base. PMID- 18391831 TI - Carcinosarcoma of the ampulla of Vater. PMID- 18391832 TI - First report of Streptococcus bovis-associated acute cholecystitis in North America. PMID- 18391833 TI - Colchicine-associated ring mitosis in liver biopsy and their clinical implications. AB - Colchicine is an alkaloid with antimitotic activity used to treat a variety of medical conditions. Colchicine toxicity can result in multiorgan failure and death. In recent years, the histopathologic features of colchicine toxicity in various organs that is, gastrointestinal tract, kidneys, salivary glands, muscle, and skin have been described. However, the histopathologic manifestations of colchicine in the liver have not been reported. We present the case of a 54-year old man with transfusion acquired chronic hepatitis C and longstanding gout managed with colchicine. A liver biopsy performed showed changes consistent with chronic hepatitis C along with many scattered mitoses arrested in metaphase in the hepatocytes that have been referred to as "ring" mitoses. The current liver biopsy was compared with the biopsy from 7 years ago, which appeared similar, however, ring mitoses were absent. Ring mitosis have been associated with colchicine-mediated tissue injury in various other sites, and their presence is felt to represent colchicine-induced liver injury in this case. PMID- 18391834 TI - Characteristics of nasal epithelial cells in naturally postmenopausal women receiving hormone therapy with 1 mg 17beta-estradiol and 2 mg drospirenone: a prospective study. AB - OBJECTIVE: To investigate the effects of drospirenone on nasal respiratory epithelium in postmenopausal women who are treated with hormone therapy (HT). DESIGN: Thirty-five naturally postmenopausal women participated in the prospective study. Women received continuous-combined HT containing 1 mg 17beta estradiol and 2 mg drospirenone. Cytological changes of nasal middle and inferior turbinate respiratory epithelium were evaluated by using the maturation index, performed at baseline and on the sixth cycle of HT. RESULTS: Hematoxylin-eosin staining for the maturation index of the epithelial nasal cells of women reached better trophic aspects during 17beta-estradiol/drospirenone intake with respect to those observed at baseline (P < 0.001). The smears during HT emphasized the superficial and the intermediate types of cells, with a ratio index greater than 2. The parabasal cell content was less than 20%, and the karyopyknotic index showed degenerative changes in superficial and intermediate cells. CONCLUSIONS: Our study confirmed that the nasal respiratory epithelium is an ovarian steroid target. Drospirenone acts on nasal cells similarly to other progestogens. PMID- 18391835 TI - Cycles within cycles. PMID- 18391836 TI - Plasma concentrations of carbohydrates and sugar alcohols in term newborns after milk feeding. AB - Nonglucose carbohydrates such as galactose, mannose, and inositol play a clinically important role in fetal and neonatal nutrition, though little is known about their metabolism in the neonate. The aim of this study was to determine whether postprandial changes in plasma carbohydrate and sugar alcohol concentrations are affected by clinical variables such as postnatal age (PNA), milk type, feeding volume, or feeding duration in term newborns. Neonates (n = 26) taking intermittent enteral feedings were enrolled. Blood samples were obtained at baseline (immediately before the start of a feeding) and at 2-3 subsequent time points up to 110 min. Postprandial rise was only observed for plasma glucose concentrations [Glu] and plasma galactose concentrations [Gal] and clinical variables did not predict this change. Despite equimolar delivery in milk, the median of [Glu] rise minus [Gal] rise from baseline to second postprandial plasma sample was 674 microM (-38, 3333 microM; p < 0.0001), reflecting efficient hepatic first-pass metabolism of galactose. A significant PNA effect on [Gal] was observed such that for each day PNA there was an 18% decrease in [Gal] (p = 0.03). [Gal] are a function of PNA, suggesting maintenance of a significant ductus venosus shunt in term infants. PMID- 18391837 TI - Natural isoprenoids are able to reduce inflammation in a mouse model of mevalonate kinase deficiency. AB - Mevalonate kinase deficiency (MKD) is a rare disorder characterized by recurrent inflammatory episodes and, in most severe cases, by psychomotor delay. Defective synthesis of isoprenoids has been associated with the inflammatory phenotype in these patients, but the molecular mechanisms involved are still poorly understood, and, so far, no specific therapy is available for this disorder. Drugs like aminobisphosphonates, which inhibit the mevalonate pathway causing a relative defect in isoprenoids synthesis, have been also associated to an inflammatory phenotype. Recent data asserted that cell inflammation could be reversed by the addition of some isoprenoids, such as geranylgeraniol and farnesyl pyrophosphate. In this study, a mouse model for typical MKD inflammatory episode was obtained treating BALB/c mice with aminobisphosphonate alendronate and bacterial muramyldipeptide. The effect of exogenous isoprenoids -- geraniol, farnesol, and geranylgeraniol -- was therefore evaluated in this model. All these compounds were effective in preventing the inflammation induced by alendronate muramyldipeptide, suggesting a possible role for these compounds in the treatment of MKD in humans. PMID- 18391838 TI - Increased elastic tissue defect formation in the growth restricted Brown Norway rat: a potential link between in utero condition and cardiovascular disease. AB - Low birth weight for gestational age has been epidemiologically linked to cardiovascular mortality and morbidity in adult life. This study aimed to determine whether in utero growth restriction influences an early feature of atherosclerotic pathology; disruption of the aortic internal elastic lamina (IEL) in the adult Brown Norway (BN) rat. In utero growth restriction was induced by bilateral uterine artery ligation on day 18 of gestation, thereby decreasing newborn BN pup weight by approximately 14%. Restriction surgery significantly increased aortic IEL defect number at 8 wk of age in both sexes compared with no surgery animals (p < 0.002). At 16 wk of age placental restriction surgery significantly increased the number of defects in males compared with both no surgery and sham surgery control groups (p < 0.001). The total number of IEL defects was significantly correlated with several postnatal growth rate parameters, including 72-h postpartum weight. Neither blood pressure was significantly different between treatment groups, nor was it correlated with body weight or IEL defect numbers. The findings of this study seem to support the fetal origins of adult disease hypothesis, by demonstrating that a moderate growth restricting insult dramatically increases aortic elastic tissue defect formation via an apparently blood pressure-independent mechanism. PMID- 18391839 TI - Aerosol delivery in ventilated newborn pigs: an MRI evaluation. AB - Pulmonary deposition of inhaled drugs in ventilated neonates has not been studied in vivo. The objective of this study was to evaluate pulmonary delivery of gadopentetate dimeglumine (Gd-DTPA) following nebulization in ventilated piglets using magnetic resonance imaging. Seven ventilated piglets (5 +/- 2 d old, weight 1.8 +/- 0.5 kg) were scanned in the Bruker/Siemens 4T magnetic resonance scanner using T1 weighted spin-echo sequence. Aerosols of Gd-DTPA were generated continuously using the MiniHeart jet nebulizer. Breath-hold coronal images were obtained before and every 10 min during aerosolized Gd-DTPA for 90 min. Signal intensity (SI) changes over the lungs, kidneys, liver, skeletal muscle, and heart were evaluated. A significant increase in SI was observed in the lungs, kidney, and liver at 10, 20, and 40 min respectively after start of aerosol. At the end of 90 min, the SI increased by 95%, 101%, and 426% over the right lung, left lung, and kidney, respectively. A much smaller increase in SI was observed over the liver. In conclusion, we have demonstrated effective pulmonary aerosol delivery within 10 min of contrast nebulization in ventilated piglets. Contrast visualization in the kidneys within 20 min of aerosol initiation reflects alveolar absorption, glomerular filtration and renal concentration. PMID- 18391840 TI - Early cardiac abnormalities in obese children: importance of obesity per se versus associated cardiovascular risk factors. AB - We investigated whether obese children and adolescents have early echocardiographic signs of subclinical cardiac dysfunction and evaluated the respective influence of obesity per se versus parameters of carbohydrate and lipid metabolism that are frequently abnormal in obese subjects. The role of tissue Doppler imaging as a screening tool for these abnormalities was explored. Blood pressure and echocardiographic parameters, including tissue Doppler measurements of the septal mitral annulus were evaluated in 49 obese children and adolescents and 45 age and sex matched controls. The respective influence of obesity versus parameters of carbohydrate and lipid metabolism was examined with linear regression analysis. Obese subjects showed significantly larger left ventricular wall dimensions (posterior wall, septum, and left ventricular mass index) and signs of early diastolic filling abnormalities on conventional and tissue Doppler echocardiography compared with nonobese subjects. Multiple regression analysis showed that mainly BMI-SD scores and/or body surface area explained significant proportions of the variance of the early cardiac abnormalities. In conclusion, young, obese children and adolescents have significant changes in left ventricular wall dimensions and early diastolic filling compared with nonobese subjects. Obesity per se and not the parameters of carbohydrate and lipid metabolism predicted the early cardiac abnormalities. PMID- 18391841 TI - The arginine-creatine pathway is disturbed in children and adolescents with renal transplants. AB - Cardiovascular disease is an important cause of morbidity in recipients of renal transplants. The aim of the present study was to analyze the status of the arginine-creatine pathway in such patients, given the relationship between the arginine metabolism and both renal function and the methionine-homocysteine cycle. Twenty-nine children and adolescents (median age 13, range 6-18 years), who had received a renal allograft 14.5-82.0 months before, were recruited for the study. On immunosuppressive therapy, all patients evidenced an adequate level of renal function. Plasma concentrations of homocysteine and glycine were significantly higher, whereas urinary excretions of guanidinoacetate and creatine were significantly lower than controls. Urinary excretions of guanidinoacetate and creatine correlated positively with creatinine clearance. Urinary excretion of creatine was negatively correlated with plasma concentration of homocysteine. The demonstration of disturbances in the arginine-creatine pathway in patients with well-functioning renal transplants and in absence of chronic renal failure represents a novel finding. We speculate that the low urinary excretion of guanidinoacetate and creatine is probably related to the nephrotoxic effect of immunosuppressive therapy and to defective methylation associated with the presence of hyperhomocysteinemia. PMID- 18391842 TI - Inflammation at birth is associated with subnormal development in very preterm infants. AB - Preterm birth carries a risk for impaired developmental outcome. We have previously described an association between increased levels of proinflammatory cytokines during the first 72 postnatal hours and cerebral damage as detected by ultrasound in a cohort of 74 very preterm infants. Sixty-seven of 71 surviving children with a mean gestational age of 27.1 (2.0) wk were examined at 2 y corrected age with a standardized neurologic examination and with Bayley Scales of Infant Development. We hypothesized that proinflammatory cytokine concentrations at or shortly after birth would be associated with an adverse developmental outcome. Increased concentrations of TNF-alpha in cord blood odds ratio (95% confidence interval) 3.3 (1.1-10.2), p = 0.013 and at 6 h 7.8 (0.9 71.8), p = 0.015 and of IL-6 in cord blood 1.7 (1.0-2.9), p = 0.048 were associated with psychomotor developmental index <85. Increased concentrations of TNF-alpha in cord blood odds ratio (95% confidence interval) 3.6 (1.002-12.8), p = 0.044 and of IL-8 in cord blood 3.5 (1.2-10.6), p = 0.023 were associated with cerebral palsy. Associations of TNF-alpha and IL-8 in cord blood with the respective outcome measures remained significant after adjustment for other clinical variables. Proinflammation at birth is associated with impaired functional outcome at 2 y of corrected age in children with very preterm birth. PMID- 18391843 TI - Bronchoalveolar lavage MMP-9 and TIMP-1 in preschool wheezers and their relationship to persistent wheeze. AB - Atopic preschool children are more likely to develop persistent wheezing, which could be a consequence of early airway remodeling. Protease-antiprotease balance between MMP-9 and its cognate inhibitor TIMP-1 may be involved in this process. Our hypothesis was that atopic wheezing preschool children would have an imbalance of MMP-9 to TIMP-1 in bronchoalveolar lavage (BAL). BAL from 52 preschool wheezers was compared with 14 controls without wheeze. A subgroup completed an International Study of Asthma and Allergy in Childhood symptom questionnaire 2 y later. Molar ratios of MMP-9/TIMP-1 were higher in wheezy children (p < 0.001; median 4.0%, range 0-8.7) than controls (0.6%, 0-1.8), and showed an excess of TIMP-1 in the airway. BAL TIMP-1 was raised in children with persistent wheezing (p = 0.028; 34.4 ng/mL, 9.1-93.1 compared with 10.6 ng/mL 6.1 18.6), as was serum levels of intercellular adhesion molecule-1 (p = 0.027). The absolute concentration of TIMP-1 in the airway, rather than its molar ratio with MMP-9, was associated with persistent wheezing. The processes involved with airway remodeling are complex but excess TIMP-1 may impede matrix protein turnover and thereby contribute to persistent changes in airway structure and wheezing. PMID- 18391844 TI - KL4-surfactant (Lucinactant) protects human airway epithelium from hyperoxia. AB - Exogenous surfactant is critical in the treatment of neonates with respiratory distress syndrome. Lucinactant (Surfaxin; Discovery Laboratories, Inc.) is a surfactant replacement therapy containing sinulpeptide, which may reduce lung inflammation. This study tested whether Lucinactant reduces markers of inflammation, damage and remodeling in human airway epithelial cells exposed to hyperoxia. Calu-3 monolayers cultured at an air-liquid interface were treated apically with 140 microL of normal saline, Lucinactant or Beractant (Survanta; Abbott Laboratories, Inc.). Treated monolayers were exposed to 60% O(2)/5% CO(2) for 24 or 72 h. Transepithelial resistance (TER; p < 0.001) and cell viability (p < 0.05) were greater in both surfactant groups compared with saline; by 72 h Lucinactant cells had greater TER than Beractant (p < 0.001). Surfactant treated groups secreted less IL-8 than saline (p < 0.001), whereas Lucinactant cells secreted less IL-6 than saline and Beractant (p < 0.001). Matrix metalloproteinase 7, expressed by saline and Beractant treated cells at 24 h, was attenuated by 72 h by Beractant (p < 0.001), but was never detected in Lucinactant cells. Histology indicated less injury with Lucinactant relative to Beractant and saline. These data suggest that Lucinactant was protective compared with Beractant and control. PMID- 18391845 TI - Development of the premature infant nose throat-model (PrINT-Model): an upper airway replica of a premature neonate for the study of aerosol delivery. AB - Clinical efficacy of aerosol therapy in premature newborns depends on the efficiency of delivery of aerosolized drug to the bronchial tree. To study the influence of various anatomical, physical, and physiological factors on aerosol delivery in preterm newborns, it is crucial to have appropriate in vitro models, which are currently not available. We therefore constructed the premature infant nose throat-model (PrINT-Model), an upper airway model corresponding to a premature infant of 32-wk gestational age by three-dimensional (3D) reconstruction of a three-planar magnetic resonance imaging scan and subsequent 3D-printing. Validation was realized by visual comparison and comparison of total airway volume. To study the feasibility of measuring aerosol deposition, budesonide was aerosolized through the cast and lung dose was expressed as percentage of nominal dose. The airway volumes of the initial magnetic resonance imaging and validation computed tomography scan showed a relative deviation of 0.94%. Lung dose at low flow (1 L/min) was 61.84% and 9.00% at high flow (10 L/min), p < 0.0001. 3D-reconstruction provided an anatomically accurate surrogate of the upper airways of a 32-wk-old premature infant, making the model suitable for future in vitro testing. PMID- 18391846 TI - Urinary prostaglandins and the effect of indomethacin on phosphate excretion in children with hypophosphatemic rickets. AB - We recently reported the urinary prostaglandin E(2)/creatinine ratio (PGE(2)/Cr) was markedly elevated in Hyp mice, the animal model for X-linked hypophosphatemia, compared with control mice. We provided evidence for altered prostaglandin production mediating the phosphaturia and that indomethacin decreases urinary phosphate excretion in Hyp mice but not control mice. To determine the levels of urinary PGE(2)/Cr, the safety and efficacy of indomethacin on phosphate excretion in children with hypophosphatemic rickets (HPR), a prospective clinical trial was performed in 16 children with HPR and 16 age- and gender-matched healthy controls. Urinary PGE(2)/Cr excretion was determined on a 24 h timed urine collection. A randomized cross over, placebo versus indomethacin, clinical trial was performed in the 16 children with HPR. There was no difference in urinary PGE(2)/Cr excretion between controls and patients with HPR. In children with HPR, indomethacin treatment for 3 mo had no significant effect on serum phosphorus or urinary phosphate excretion. In conclusion, urinary prostaglandin excretion is similar in children with HPR compared with controls. Indomethacin had no significant effect on serum phosphorus or urinary phosphate excretion in children with HPR. PMID- 18391847 TI - Genetic analysis of congenital cystic adenomatoid malformation reveals a novel pulmonary gene: fatty acid binding protein-7 (brain type). AB - The pathogenesis of congenital cystic adenomatoid malformation (CCAM) is unknown and its natural history is unpredictable. Fatty acid binding protein-7 (FABP-7) has been previously described in brain and breast development, but never before in the lung. We investigate gene expression in CCAM, and hypothesize that CCAM results from an aberration in the signaling pathway during lung development. Under IRB approval, tissue specimens of fetal CCAM, fetal control, postnatal CCAM, and postnatal control were examined and microarray analysis was performed. Candidate differentially expressed genes were selected with log-odds ratio (B) >0 and false discovery rate <0.05. Validation of differential expression was achieved at the RNA and protein levels. FABP-7 was underexpressed in fetal CCAM compared with fetal lung in both the microarray and by RT-PCR. Findings were duplicated by Western Blot analysis and immunohistochemistry. This is the first description of FABP-7 in the human lung. Decreased expression of FABP-7 in fetal CCAM compared with normal fetal lung at both the RNA and protein levels suggests FABP-7 may have a role in pulmonary development and in the pathogenesis of CCAM. PMID- 18391848 TI - Delayed hypothermia as selective head cooling or whole body cooling does not protect brain or body in newborn pig subjected to hypoxia-ischemia. AB - The neuroprotective efficacy of hypothermia (HT) after hypoxia-ischemia (HI) falls dramatically the longer the delay in initiating HT. Knowledge is scarce regarding protective or adverse effects of HT in organs beyond the brain. In addition, the relative effectiveness of selective head cooling (SHC) and whole body cooling (WBC) has not been studied. We aimed to examine whether 24 h HT, initiated 3 h after global HI is brain- and/or organ-protective using pathology, neurology, and biochemical markers. Fifty, of 10). Relaxation improved more with music than rest (p < 0.01). The HRV parasympathetic parameter was significantly lower with music than rest. No other differences were significant. Further studies are needed to better delineate the relationship between subjective and objective measures of well-being among patients who are not in severe distress. PMID- 18391851 TI - Bone mineral content at birth is determined both by birth weight and fetal growth pattern. AB - Adult peak bone mass is related to birth weight, suggesting it could be affected by fetal growth pattern. Small-for-gestational-age (SGA) newborns have lower bone mineral content (BMC), but what about adapted-for-gestational-age (AGA) newborns with fetal growth restriction? The purpose of the study was to determine the respective role of birth weight and fetal growth pattern on BMC. Full-term newborns from SGA high-risk pregnancies were included (n = 185). Estimated fetal weight percentiles were measured monthly from mid-gestation to birth, and restricted fetal growth (FGR) was defined as a loss by more than 20 percentiles. BMC was measured at birth, using dual x-ray absorptiometry. Newborns were SGA (n = 56) or AGA (n = 129). Newborns with FGR (n = 111) were AGA (n = 71) or SGA (n = 41). BMC was significantly lower in SGA than AGA (1.48 +/- 0.02 vs. 1.87 +/- 0.04 g/cm) and lower when FGR irrespective of birth weight (1.66 g/cm +/- 0.03 vs. 1.89 g +/- 0.05). In multivariate analysis, FGR and SGA were significant and independent predictors of low BMC. In conclusion, fetal growth pattern affects BMC not only in SGA infants but also when birth weight is maintained in the normal range. PMID- 18391852 TI - Nitric oxide suppresses the secretion of vascular endothelial growth factor and hepatocyte growth factor from human mesenchymal stem cells. AB - The production of growth factors such as vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) by human bone marrow mesenchymal stem cells (MSCs) may play an important role in their paracrine effects on proliferation, differentiation, and protection. NO is produced during ischemia and may affect MSC function. However, it is unknown whether NO alters the production of VEGF and HGF from MSCs. To study this, human MSCs were stimulated to produce growth factors with TNF or LPS with and without various doses of NO donors or NOS inhibitors. We found that FK409, an NO donor, significantly suppressed the production of VEGF and HGF from human MSCs. Vascular endothelial growth factor in the supernatants of cells treated by 20 nM FK409 (497 +/- 19 pg/mL) was significantly lower compared with controls (625 +/- 34 pg/mL). Similarly, NO donor significantly suppressed the amount of HGF from controls (118 +/- 3 to 40 +/- 2 pg/mL) after treatment with 20 nM FK409. NO donor also abolished the augmentation of VEGF production induced by LPS. The amount of VEGF in the supernatant was 571 +/- 11 pg/mL when cells were treated with 20 nM FK409 and LPS (200 ng/mL), which was significantly lower than groups treated with LPS alone (941 +/- 30 pg/mL). This study constitutes an initial report regarding the effect of NO on human MSC growth factor production. PMID- 18391853 TI - Somatostatin infusion increases intestinal ischemia and does not improve vasoconstrictor response to norepinephrine in ovine endotoxemia. AB - Hemodynamic support of patients with septic shock is often complicated by a tachyphylaxis against exogenous catecholamines. Because an increase in somatotropic hormones may play a pivotal role in the regulation of the inflammatory response to endotoxin, intravenous supplementation of the neuroendocrine hormone somatostatin (SOMA) may attenuate hemodynamic dysfunction resulting from endotoxemia. The objective of the present study was to assess the short-term effects of SOMA alone and in combination with norepinephrine (NE) on cardiopulmonary hemodynamics, global oxygen transport, plasma nitrate/nitrite levels, and intestinal integrity compared with single NE therapy in ovine endotoxemia. After a baseline measurement in healthy sheep (n = 16) had been performed, Salmonella typhosa endotoxin was centrally infused (10 ng x kg(-1) x min(-1)) to induce a hypotensive-hyperdynamic circulation using an established protocol. Animals surviving 16 h of endotoxemia were randomly assigned to one of the two groups (each n = 6). Sheep allocated to the SOMA + NE group received SOMA as a loading dose of 10.5 microg x kg(-1) x min(-1) for 1 h, followed by a continuous infusion of 3.5 microg x kg(-1) x min(-1) for the next 2 h. After the SOMA loading dose had been given, NE was concurrently infused (0.3 microg x kg( 1) x min(-1)) for 2 h. In the NE group (control), NE (0.3 microg x kg(-1) x min( 1)) was continuously infused for 3 h. Endotoxemia caused a decrease in MAP and systemic vascular resistance index in both groups, but to a greater extent in the NE group. Arterial hypotension persisted despite administration of the study drugs. Infusion of SOMA alone and in combination with NE did not significantly increase systemic vascular resistance index. Neither SOMA nor NE infusion alone affected pulmonary vasoregulation. Plasma nitrate/nitrite levels did not differ between groups. However, combined infusion of SOMA and NE significantly increased arterial lactate concentrations, oxygen consumption index, and oxygen extraction rate (P < 0.05) and aggravated ileal mucosal injury. In conclusion, short-term treatment with SOMA failed to attenuate cardiocirculatory shock resulting from endotoxemia and did not improve vasopressor response to NE. In addition, combined SOMA and NE therapy resulted in intestinal injury. Therefore, SOMA does not seem to represent a therapeutic option to treat arterial hypotension resulting from sepsis and systemic inflammatory response syndrome. PMID- 18391854 TI - Systematic analysis of the salutary effect of estrogen on cardiac performance after trauma-hemorrhage. AB - Although 17beta-estradiol (estrogen) and estrogen receptor (ER) agonist administration after trauma-hemorrhage improves cardiac function, it remains unknown what the optimal estrogen or ER agonist dosage is to elicit this beneficial effect. To study this, the dose-dependent effects of estrogen, propylpyrazole triol (ER-alpha agonist), and diarylpropionitrile (DPN; ER-beta agonist) on heart performance (+dP/dt) were determined in sham rats and in experimental animals at the time of maximal bleedout (MBO) or at 2 h after trauma hemorrhage. The results showed that estrogen and DPN induced dose-dependent increases in the maximal rate of left ventricular pressure increase (+dP/dt) in all groups, whereas propylpyrazole triol was ineffective at all doses. The maximal dose and the 50% effective dose of DPN were approximately 100-fold lower than those of estrogen. The half-life of estrogen in plasma was approximately 25 min in sham and MBO groups. A positive correlation between the estrogen-induced increase in +dP/dt and survival in MBO rats were observed. These results collectively suggest that the salutary effects of estrogen on cardiac performance are dose-dependent and mediated via ER-beta. PMID- 18391855 TI - Characterization of the inflammatory response during acute and post-acute phases after severe burn. AB - Severe burn causes a pronounced hypermetabolic response characterized by catabolism and extensive protein wasting. We recently found that this hypermetabolic state is driven by a severe inflammatory response. We characterized in detail the kinetics of serum levels of a panel of cytokines in a rat model, which may serve as reference for the development of therapeutic interventions applicable to humans. Male Sprague-Dawley rats (n = 8) received a full-thickness burn of 60% total body surface area. Serum was harvested 1, 3, 6, 12, 24, 48, 96, and 168 h after burn. Eight serum cytokines commonly used to assess the inflammatory response in humans, such as IL-1beta, IL-6, IL-10, TNF, vascular endothelial growth factor, and monocyte chemotactic protein 1, and the rat-specific cytokines cytokine-induced neutrophil chemoattractant (CINC) 1, CINC 2, and CINC-3 were measured by enzyme-linked immunosorbent assay technique and were compared with controls (n = 4). Statistical analysis was conducted using the t test, with P < 0.05 considered as significantly different. Thermal injury resulted in significantly increased serum levels of IL-1beta, IL-6, IL-10, monocyte chemotactic protein 1, CINC-1, CINC-2, and CINC-3 when compared with the concentrations detected in nonburned rats (P < 0.05). Serum levels of TNF-alpha and vascular endothelial growth factor in burned rats were not found to be significantly different to controls. Burn causes a profound inflammatory response in rats. Specific cytokines known to increase in humans postburn such as IL-1 beta, IL-6, IL-10, MCP-1, and IL-8 (CINC-1, CINC-2, and CINC-3 in the rat) were also observed in our rat burn model, which now allows us to study new anti inflammatory treatment options. PMID- 18391856 TI - Human cholesteryl ester transfer protein expression enhances the mouse survival rate in an experimental systemic inflammation model: a novel role for CETP. AB - Mice expressing human cholesteryl ester transfer protein (huCETP) are more resistant to Escherichia coli bacterial wall LPS because death rates 5 days after intraperitoneal inoculation of LPS were higher in wild-type than in huCETP+/+ mice, whereas all huCETP+/+ mice remained alive. After LPS inoculation, plasma concentrations of TNF-alpha and IL-6 increased less in huCETP+/+ than in wild type mice. LPS in vitro elicited lower TNF-alpha production by CETP expressing than by wild-type macrophages. In addition, TNF-alpha production by RAW 264.7 murine macrophages increased on incubation with LPS but decreased in a dose dependent manner when human CETP was added to the medium. Human CETP in vitro enhanced the LPS binding to plasma high-density lipoprotein/low-density lipoprotein. The liver uptake of intravenous infused 14C-LPS from Salmonella typhimurium was greater in huCETP+/+ than in wild-type mice. Present data indicate for the first time that CETP is an endogenous component involved in the first line of defense against an exacerbated production of proinflammatory mediators. PMID- 18391857 TI - Depletion of NK cells in a murine polytrauma model is associated with improved outcome and a modulation of the inflammatory response. AB - Sepsis and associated diseases such as systemic inflammatory response syndrome and multiple organ dysfunction syndrome represent common posttraumatic complications on intensive care units induced by a variety of body defense mechanisms. Natural killer (NK) cells are part of the innate immune system. They are thought to play an important role in the development of such syndromes by interplay with other immune cell types and subsequent activation of the inflammatory cascade. To test this hypothesis, NK cells were depleted by administration of antimouse asialo-GM1 antibody in a murine polytrauma model consisting of femur fracture, hemorrhagic shock, and subsequent sepsis. Mortality and immune parameters such as cytokine expression in lung and liver, lymphocyte phenotyping, lymphocyte apoptosis, and organ pathology were determined 96 h after sepsis induction. Survival values showed 50% in the control sepsis group and 100% after NK cell depletion. Thus, NK cell depletion resulted in 50% mortality reduction. Furthermore, we found reductions in the inflammatory response, represented by IL-6 expression in liver, and a decrease in infiltrating neutrophils in the liver and lung. In addition, lymphocyte apoptosis in spleen was decreased by depletion of NK cells. Taken together, these data demonstrate that NK cells contribute to the pathogenetic pathways in a murine polytrauma model. One main mechanism of action seems to be the induction of systemic inflammatory events. Thus, depletion of NK cells results in attenuated inflammation and an overall improvement in outcome. Therefore, NK cells can be considered as important targets for therapeutic strategies. PMID- 18391858 TI - The protective effect of the cholinergic anti-inflammatory pathway against septic shock in rats. AB - To investigate the effects of the cholinergic anti-inflammatory pathway on hemodynamics, blood biochemistry, the plasma TNF-alpha level, and the nuclear factor-kappaB (NF-kappaB) activation during septic shock, male Sprague-Dawley rats were subjected to cecal ligation and puncture (CLP, a model of polymicrobial sepsis) or sham operation. Forty-eight rats were randomly assigned into six equal groups: sham CLP group; CLP group; VGX group was subjected to bilateral cervical vagotomy after CLP; STM group was subjected to bilateral cervical vagotomy after CLP plus the left vagus nerve trunk electrical stimulation; THA group was administered tetrahydroaminoacridine after CLP and bilateral cervical vagotomy; and alpha-BGT group was administered alpha-bungarotoxin before electrical stimulation of the vagus nerve. The right carotid artery was cannulated to monitor MAP. The plasma TNF-alpha level was measured using enzyme-linked immunosorbent assays. The hepatic NF-kappaB activation was determined by Western blotting. Cecal ligation and puncture produced progressive hypotension. Serum aspartate transaminase and alanine transaminase levels significantly increased after CLP challenge. The plasma TNF-alpha level and the hepatic NF-kappaB activation significantly increased after CLP alone or with bilateral cervical vagotomy compared with sham-operated group. Application of constant voltage pulses to the caudal vagus trunk significantly prevented the development of CLP induced hypotension, alleviated the hepatic damage, and reduced the plasma TNF alpha production, but electrical stimulation had no effect on the hepatic NF kappaB activation. Tetrahydroaminoacridine administration after bilateral cervical vagotomy reversed hypotension and attenuated the plasma TNF-alpha response; in addition, it had no effect on the hepatic NF-kappaB activation. alpha-Bungarotoxin pretreatment significantly reversed the inhibitory effect of vagal electrical stimulation, but it had no effect on the hepatic NF-kappaB activation. Our results showed that the cholinergic anti-inflammatory pathway might produce a potential protective effect on polymicrobial sepsis in rats. PMID- 18391859 TI - Efficacy of LL-37 and granulocyte colony-stimulating factor in a neutropenic murine sepsis due to Pseudomonas aeruginosa. AB - A promising therapeutic strategy for the management of severe Pseudomonas infection in neutropenic patients may result from the coadministration of colony stimulating factors (CSFs) that help maintain immune competence and antimicrobial peptides, a novel generation of adjunctive therapeutic agents with antimicrobial and anti-inflammatory properties. A promising peptide with these properties is LL 37, the only member of the cathelicidin family of antimicrobial peptides found in humans. BALB/c male mice were rendered neutropenic by intraperitoneal administration of cyclophosphamide on days -4 and -2 preinfection. Septic shock was induced at time 0 by intraperitoneal injection of 2x10 colony-forming units of P. aeruginosa American Type Culture Collection (ATCC) 27853. All animals were randomized to receive intravenously isotonic sodium chloride solution, 1 mg/kg of LL-37, 20 mg/kg of imipenem, 0.1 mg/kg of granulocyte CSF (G-CSF), 1 mg/kg of LL 37+0.1 mg/kg of G-CSF, or 20 mg/kg of imipenem+0.1 mg/kg of G-CSF. Lethality and bacterial growth in blood, peritoneum, spleen, liver, and kidney were evaluated. All regimens were significantly superior to controls at reducing the mouse lethality rate and bacterial burden in organs. Particularly, the combination between LL-37 and G-CSF was the most effective in protecting neutropenic mice from the onset of sepsis and in vitro significantly reduced the apoptosis of neutrophils. Combination therapy between LL-37 and G-CSF is a promising therapeutic strategy for the management of severe Pseudomonas infection complicated by neutropenia. PMID- 18391861 TI - Alpha-defensin-like product and asymmetric dimethylarginine increase in mesenteric lymph after hemorrhage in anesthetized rat. AB - Mesenteric lymph contains unidentified proinflammatory mediators that increase in concentration after hemorrhage. In the search for candidate mediators, we examined mesenteric lymph for the presence of proinflammatory substances that are known to be produced in the gut: (a) antimicrobial peptides and antimicrobial proteins produced in the Paneth cells of the intestine (alpha-defensin 4, secretory phospholipase A2 [sPLA2], and Reg 2 protein) and (b) asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NOS. Anesthetized male rats were hemorrhaged to 40 mmHg and maintained at that pressure by intermittent blood withdrawal until the pressure fell to less than 40 mmHg (decompensation) at which point they were resuscitated with three times the shed blood volume of Ringer's lactate solution administered over 1 h. Mesenteric lymph samples were analyzed for ADMA by enzyme-linked immunosorbent assay and for alpha-defensin 4, sPLA2, and Reg2 by Western blotting. Protein concentration in lymph was unchanged by hemorrhage, but alpha-defensin 4 increased significantly (12-fold greater than control) as did ADMA (2-fold greater than control). The sPLA2 could not be detected in lymph, and Reg 2 was unchanged during hemorrhage. During resuscitation, lymph flow tended to increase, but the concentration of ADMA and alpha-defensin 4 by volume did not increase. Reg 2 decreased during resuscitation. The results indicate that ADMA and immunoreactive product to alpha defensin 4 may contribute to the increase in inflammatory activity of mesenteric lymph during hemorrhage, but they are unlikely to be the mediators responsible for the increase in the concentration of inflammatory mediators in postresuscitation lymph. PMID- 18391862 TI - From Archives des Maladies du Coeur et des Vaisseaux to Archives of Cardiovascular Diseases: changing format for improved content. PMID- 18391863 TI - Archives of Cardiovascular Diseases: a demanding challenge. PMID- 18391864 TI - Archives of Cardiovascular Diseases: objectives and commitments. PMID- 18391865 TI - Stenting for unprotected left main coronary artery stenosis: a further step. PMID- 18391866 TI - Evaluation of aortic stiffness to predict and prevent the risk of atrial fibrillation in hypertensive patients in their 50's. PMID- 18391868 TI - Wolff-Parkinson-White syndrome in the elderly: clinical and electrophysiological findings. AB - BACKGROUND: Screening for Wolff-Parkinson-White (WPW) syndrome is recommended in children and young adults. The aim of this study was to evaluate the clinical and electrophysiological characteristics of patent WPW syndrome in subjects > or =60 years of age. METHODS: Four-hundred and fifty-nine consecutive patients with WPW syndrome, aged 8-80 years, were recruited; 32 (7%) of these patients were > or =60 years of age. The clinical, electrophysiological and therapeutic data for these patients were evaluated. RESULTS: Sixteen men and 16 women, aged 60-81 years (67+/-4.5), were admitted for resuscitated sudden death (1), rapid atrial fibrillation (4), syncope (4), or junctional tachycardia (13); 10 patients were asymptomatic (10). Left lateral bundles of Kent were detected more frequently in patients over 60 years (56%) than in those<60 years of age (40.5%). Reciprocal tachycardia was induced in 58% of subjects<60 years of age and 53% of those > or =60 years old (difference not significant); atrial fibrillation was more frequent in subjects > or =60 years of age (37.5% vs. 19%) (p<0.05). The incidence of malignant forms of WPW syndrome was identical in older and younger subjects. Ablation of the accessory pathway was indicated 18 times; effective ablation of a left bundle of Kent required a second intervention more often in patients > or =60 years of age (22% vs. 5%) (p<0.05). CONCLUSION: WPW syndrome is not uncommon in subjects over 60 years of age (7%). Left lateral accessory pathways, that have similar conduction properties to those in much younger subjects, are common. Ablation of the bundle of Kent is often difficult but is indicated in symptomatic subjects or those with more serious forms of WPW syndrome. PMID- 18391867 TI - Clinical and angiographic results of angioplasty with a paclitaxel-eluting stent for unprotected left main coronary artery disease (a study of 101 consecutive patients). AB - BACKGROUND: After coronary stenting with drug eluting stents, long-term clinical outcome of unprotected left main coronary artery disease is unknown, even large scale registries or randomised trials with coronary artery bypass graft are ongoing. AIMS: To report clinical and angiographic results of paclitaxel-eluting stent implantation for left main coronary artery stenosis (a series of 101 consecutive patients). METHODS: This report is a prospective study performed to evaluate the immediate and mid-term clinical and angiographic outcomes of patients undergoing paclitaxel-eluting stent (PES) implantation for unprotected left main coronary artery (LMCA) stenosis. From January 2004 to December 2005, 101 consecutive patients were stented with paclitaxel-eluting stents (the provisional T stenting technique followed by Kissing balloon for distal left main vessel disease). RESULTS: Mean age was 68.9+/-11.07 years. 73.3% of patients were male. Acute coronary syndrome was present in 65% of patients, of whom 22.8% had ST elevation. Distal left main trunk lesions were present in 87.1% of cases. Three-vessel disease represented 7% of cases. Angiographic success was obtained in 97.03% of patients with an acute gain of 2.18+/-0.53mm. GpIIbIIIa inhibitors were used in only 8.9% of cases. Hospital stay was 7.6 +/- 3.7 days. In-hospital complications were present in 7.9%, with a hospital mortality rate of 2%. At six month follow-up, the rate of target lesion revascularization (TLR) was 3%, and the rate for major adverse cardiac events (MACE) was 8.9%. Angiographic control was performed in 88.1% and a late loss of 0.1mm (0.04-0.2mm) was noted. Re stenosis occurred in 4 patients (4.5% of cases). 4 patients (4%) died, including 2 from cardiac causes. CONCLUSION: Paclitaxel-eluting stent implantation for unprotected left main coronary disease appears to be safe with high procedural success rate and a low re-stenosis rate at six month-follow-up. PMID- 18391869 TI - Particularities of peripheral arterial disease managed in vascular surgery in the French West Indies. AB - BACKGROUND: Epidemiology of peripheral arterial disease is currently unknown in French West Indies (Antilles). AIMS AND METHODS: The aim of this study is to present peripheral arterial disease (PAD) occurring in the French West Indian subjects through the analysis of our database of vascular surgery. The study population included 754 patients (mean age 73 +/- 10 years), mostly from African descents. The main clinical presentation was critical limb ischemia (66%, including tissue loss in 48% of cases), followed by claudication (20%). The lesions affected the infragenicular arteries in 86% of cases, including 24% isolated to this level as well as 51% combined to femoro-popliteal lesions and only 7% of cases affecting the aorto-iliac level. RESULTS: Ankle-brachial index was at 0.57 +/- 0.13 and 0.34 +/- 0.22 (p<0.001) in patients with claudication and critical limb ischemia (CLI), respectively. The severity scores were significantly higher in claudicants with aorto-iliac disease and CLI patients with infragenicular lesions. Except for hypertension (85%) and obesity (19%), the other risk factors were differently distributed between the 2 groups. While in the CLI group, patients were older, with higher rates of female sex, diabetes (62% vs. 48%, p<0.001) and severe renal failure, claudicants were significantly younger, with higher rates of smokers among men (75% vs. 51%, p<0.001) and moderate dyslipidemia (52% vs. 36%, p<0.001). The association with carotid stenosis (12%) and ischemic heart disease (18%) were quite uncommon. Renal disease (glomerular filtration rate<60 ml/mn/1.73 m2) was present in 61% of cases. CONCLUSION: This study highlights clear differences regarding the presentation, localization and associations of PAD in the West Indies subjects managed in vascular surgery, especially with a severe infragenicular disease, even in claudicants. This study suggests the effect of a different distribution of risk factors as well as other ethnic and socio-economic factors. PMID- 18391870 TI - Mechanical circulatory assistance in myocardial infarction with refractory cardiogenic shock: clinical experience in 10 patients at a teaching hospital in Rouen. AB - BACKGROUND: In patients with acute myocardial infarction (MI), cardiogenic shock (CS) remains associated with a high mortality (close to 50%) despite optimal therapeutic strategy. For those patients who are unlikely to survive, mechanical circulatory support (MCS) might be an additional life saving strategy. OBJECTIVE: To evaluate the efficacy of circulatory assistance in myocardial infarction complicated by cardiogenic shock. METHODS: We retrospectively studied the characteristics and clinical outcome of 10 patients hospitalized with acute MI and CS who required MCS. Mean age was 52+/-8 years; location of MI was anterior in 80% of cases. Immediate coronary angiography was performed in all cases 5.8+/ 7.0 hours from the onset of symptoms. Intra-aortic balloon pumping was used in 70% of patients and 30% received thrombolysis. Angioplasty with stent implantation was performed in 8 patients. RESULTS: In all patients MCS was placed within a mean of 57+/-92 hours after admission for hemodynamic instability (systolic aortic pressure: 85+/-13 mmHg; mean: 64+/-10 mmHg). Extracorporeal membrane oxygenation (ECMO) was implanted in 8 patients followed by Thoratec in one. The other 2 patients received a Thoratec and a Heartmate II system respectively. Survival rate was 40% (4 patients): 3 patients underwent heart transplantation at a mean of 93+/-97 days and one patient is alive with definitive implantable Heartmate. The other six patients died in hospital. CONCLUSION: Mechanical circulatory support appeared life saving in 4 out of 10 patients with acute MI and cardiogenic shock refractory to optimal treatment. In this situation, circulatory assistance deserves discussion and the choice of optimal device should be further evaluated. PMID- 18391871 TI - Arterial stiffness is associated with left atrial size in hypertensive patients. AB - BACKGROUND: Arterial stiffness is a strong predictor of cardiovascular events and particularly of stroke. A likely explanation is the development of atherosclerotic lesions at the carotid level, favored by increased local stiffness. Another possibility involves cardiac consequences of aortic stiffness and particularly left atrial dilatation with its subsequent risk of atrial fibrillation (AF) and cerebral embolism. AIMS: The present study investigated the link between arterial stiffness, pulse pressure and left atrial size, a determinant of AF risk. METHODS: Arterial stiffness was determined from pulse wave velocity (PWV) and pulse pressure (PP). Left atrial size was also measured. Several potential confounders were taken into account including indices of ventricular remodeling and diastolic function (estimated by NT-Pro brain natriuretic peptide (NT-proBNP) levels). RESULTS: Three-hundred and ten hypertensive patients, aged 53 +/- 13 years, were included. Mean 24-h blood pressure (BP) was 154 +/- 20 over 93 +/- 13 mmHg. Significant relationships were found between left atrial diameter (LAD) and PWV (r=0.27, P<0.001) and between LAD and 24-h PP (r=0.32, P<0.001). LAD was also correlated significantly, although not always tightly, with left ventricular dimensions, geometry and NT proBNP. In two different multivariate models, LAD remained significantly correlated with PWV or with 24-h PP, independently of classical determinants like age, gender, body mass index, ventricular remodeling (i.e. dimensions and geometry) and filling pressure. CONCLUSION: These results led us to propose AF as a new possible pathophysiological link between arterial stiffness and stroke. These results also emphasize the cardiac consequences of arterial stiffness which can fuel a new approach to AF prevention. PMID- 18391873 TI - Effect of time to treatment and age on one year mortality in acute STEMI: difference between thrombolysis and primary percutaneous coronary intervention. AB - CONTEXT: Although thrombolysis (THL) and primary percutaneous coronary intervention (PPCI) are two validated options in reperfusion algorithms for ST elevation myocardial infarction (STEMI), recent papers seems to show that PPCI could be the best therapeutic option irrespective of the time to treatment (TT) and of the cardiovascular risk profile of the patient. The impact of TT and age on reperfusion strategies requires elucidation. The aim of this study was to analyze the effect of time to treatment and age on the 1-year mortality of patients presenting with STEMI, for each reperfusion strategy. MATERIALS AND METHODS: The study population consisted in 794 patients directly admitted to the cardiological intensive care unit for STEMI < or =12 hours. The relationship between TT and 1-year mortality was studied using logistic regression models. The models were implemented on the overall population and on 3 different age groups: [<65 years]; [> or =65 and <75 years]; [> or =75 years] for patients undergoing THL (n=299) and for patients undergoing PPCI (n=495). There was no significant between-group difference in all-cause 1-year mortality for the patients [<65 years] and those [> or =65 and <75 years]. In contrast, the 1-year mortality was significantly higher in the patients [> or =75 years] undergoing THL (51.4 vs. 15.3%; p<0.001). The analysis of the curves of mortality suggests that 1- year mortality of patients with STEMI depend not only on reperfusion strategy but so on the time to treatment and on the age of the patients. CONCLUSION: In STEMI, on a 1-year mortality criteria, PPCI is not always upper than THL, particularly for patients<65 years treated within the first two hours after symptoms onset. TT and age affects the results of the reperfusion strategies and must be still incorporated in the reperfusion algorithms of STEMI. PMID- 18391874 TI - Cardiac resynchronization therapy: which device to implant? AB - Cardiac resynchronization therapy is now a validated treatment for patients with moderate to severe heart failure despite optimal drug treatment with left ventricular systolic dysfunction and cardiac dyssynchrony defined by wide QRS greater than 120 ms. Once an indication for cardiac resynchronization therapy has been confirmed the choice of the most appropriate device (pacemaker or intracardiac cardioverter defibrillator (ICD)) needs to be made. In heart failure patients the risk of sudden death, mainly but not always related to arrhythmic cause is high. Previous studies of primary prevention of sudden cardiac death in patients with a poor left ventricular function have shown that ICD therapy significantly reduces overall mortality and arrhythmic mortality. However patients candidates to cardiac resynchronization therapy are different from those included in the ICDs trials because they are older and have more comorbidities. The choice of the devices has to consider the potential benefit of the therapy, the comorbidities, the life expectancy but also the cost-effectiveness and the potential complications related to the device. Now, new devices provide information about the hemodynamic status of this heart failure population and thus provide an early detection of heart failure decompensation. The development of home monitoring should alert very early the physicians of the occurrence of a heart failure decompensation and thus to avoid recurrent hospitalisations for heart failure decompensation. PMID- 18391875 TI - Assessment of myocardial ischemia and viability using tissue Doppler and deformation imaging: the lessons from the experimental studies. AB - Tissue Doppler imaging and strain rate imaging are quantitative methods for assessing myocardial function and have been shown to overcome the limitations of current ultrasound methods in assessing the complex changes in regional myocardial function that occur in differing ischemic substrates. Tissue Doppler imaging (TDI) measures in real time the myocardial velocity gradient which is an index of myocardial deformation. Strain and strain rate (SR) imaging has been shown to be a sensitive technique for quantifying regional myocardial deformation. Strain rate is less load-dependent that strain and provides therefore a better measure of contractility. In the setting of ischemia, experimental studies have shown that strain imaging was an accurate method for quantitative evaluation of regional myocardial function and may yield important physiological data. In myocardial infarction, transmural extension of scar distribution in the infarct zone is proportionally related to the reduction in systolic function measured by the radial transmural velocity gradient or by strain rate imaging. Measurement of both systolic and post-systolic deformation both at rest and during a graded dobutamine infusion may help to distinguish between transmural and non transmural infarcts. In conclusion, strain imaging has the ability to evaluate of regional myocardial function. Strain rate has not replaced conventional grey-scale imaging in the assessment of regional left ventricular function and the implement of these new indices in the routine clinical practice will need additional clinical and large-scale studies. PMID- 18391876 TI - Secundum atrial septal defect assessed by 2D and 3D echocardiography in an adult: atrial septal defect and 3D echo. PMID- 18391872 TI - Echocardiographic assessment of the incidence of mechanical complications during the early phase of myocardial infarction in the reperfusion era: a French multicentre prospective registry. AB - BACKGROUND: Since the early reports on the incidence of mechanical complications of acute myocardial infarction (AMI) assessed by echocardiography published in the 1980s, the management of patients with AMI has changed considerably, in particular with the progressive development of early revascularisation. METHODS: The aim of this multicentre study was to assess the incidence of mechanical complications of AMI in the reperfusion era. Nine-hundred and eight consecutive patients were included. Echocardiography was performed on admission and at discharge. Seventy-eight percent of patients were revascularised at the acute phase. RESULTS: The following incidence rates of mechanical complications were observed: mitral regurgitation 28%, secondary to left ventricular (LV) remodelling (43%) or papillary muscle dysfunction (57%); pericardial effusion 6.6%, more frequent after anterior AMI and associated with a lower ejection fraction (EF); LV thrombus 2.4%, mainly after anterior AMI and associated with a lower EF (38+/-10% vs. 48+/-12%; p<0.001); early infarct expansion 4%; septal rupture 0.6%; and acute free wall rupture 0.8%. The following factors were independently associated with the occurrence of mechanical complications by multivariate logistic regression analysis: lack of early revascularisation (OR 3.48, 95%CI 1.36-8.95; p<0.001), LV-EF<50% (OR 1.95, 95%CI 1.42-2.67; p<0.001), Killip class>II (OR 1.91, 95%CI 1.27-2.87; p<0.002) and age > or =70 years (OR 1.42, 95%CI 1.03-1.97; p<0.03). CONCLUSION: This study demonstrates the favourable prognostic influence of early revascularisation as shown by the low incidence of mechanical complications after AMI, and underlines the persistent relationship between the development of these complications and depressed LV function. PMID- 18391877 TI - Nearly circumferential delayed contrast enhancement in severe ischemic cardiomyopathy. PMID- 18391878 TI - 2D strain imaging in the setting of acute myocardial infarction. PMID- 18391879 TI - Make the most of your doctor visit. A checklist for your checkup. PMID- 18391880 TI - Hysterectomy may increase risk of urinary incontinence. PMID- 18391881 TI - Delay in reaching hospital after a heart attack starts affects survival. PMID- 18391882 TI - Constipation. Is there help? PMID- 18391883 TI - Mayo Clinic office visit. CT scans and radiation. An interview with Richard Vetter, Ph.D. PMID- 18391885 TI - How do I know if it's just heartburn or something more serious? PMID- 18391884 TI - Low-calorie sweeteners. Every bit as sweet, but are they safe? PMID- 18391886 TI - Recently, a walk-in clinic was added to a nearby retail store. Are these clinics safe to use? PMID- 18391887 TI - Importance of understanding what is happening with HIV in different body compartments. PMID- 18391888 TI - HIV-1 in the male genital tract: evidence and impact of compartmentalisation. PMID- 18391889 TI - HIV in the lymphoid compartment. PMID- 18391890 TI - Clinical significance of central nervous system as sanctuary site for HIV. PMID- 18391891 TI - New horizons in antiretroviral drug resistance. PMID- 18391892 TI - Epidemiology of antiretroviral resistance and impact on treatment outcomes. PMID- 18391893 TI - Resistance testing methodologies and mechanisms of resistance. PMID- 18391894 TI - Learning from HIV: how will drug resistance influence treatment strategies against hepatitis C? PMID- 18391896 TI - Resistance to integrase inhibitors. PMID- 18391895 TI - Clinical implications of resistance patterns against HBV-polymerase inhibitors for the treatment of chronic hepatitis B. PMID- 18391897 TI - In brief: Meningococcal prophylaxis. PMID- 18391898 TI - Simcor: a niacin/simvastatin combination. PMID- 18391899 TI - Nilotinib (Tasigna) for CML. PMID- 18391900 TI - Tadalafil (Cialis) once a day for erectile dysfunction. PMID- 18391901 TI - Anatomy of the posterior cruciate ligament. AB - The views on the structure of the posterior cruciate ligament presented in the literature are not uniform. They can be ranged from those that refer to the ligament as an indivisible structure to those that regard it as an increasingly complex structure consisting of a number of bundles. The divisions are based on criteria related to the attachments, arrangement and functions of the components of the posterior cruciate ligament. Some researchers also specify the role of the receptors of the ligament. PMID- 18391902 TI - Modified Colzi method in the management of established Volkmann contracture--the experience of Trzebnica Limb Replantation Center (preliminary report). AB - BACKGROUND: "The terrible disability of irreversible ischaemic flexion contracture of the metacarpi and digits known as Volkmann's contracture can be avoided in the vast majority of cases, but never completely cured" (M. Koszla). A modification of Colzi's forgotten technique for the treatment of Volkmann's contracture is presented together with our treatment outcomes. MATERIAL AND METHOD: The Colzi procedure was performed in 15 patients aged: 3-54 years with Buck-Gramcko grade III, III/IV and IV Volkmann's contractures. Time between injury and surgery ranged from 4.5 months to 21 years. The main manoeuvre to reduce the contracture was two-level shortening osteotomy in the proximal and distal thirds of the radius and ulna. Osteosynthesis was performed by Rush intramedullary nailing. RESULTS AND CONCLUSIONS: A very good range of mobility (from 17 degrees preop. to 56 degrees postop.), force (0.73 preop.- 7.6 kg postop.) and contracture correction (183 degrees preop. - 27 degrees postop.) was achieved. Complications in the form of delayed union or secondary fractures occurred in 4/15 patients, and were managed by prolonged immobilisation and decortication with bone grafting. The modified Colzi procedure proved its value as a universal, easy, minimally invasive and safe method of treatment of Volkmann's contracture regardless of patient age, duration of illness, previous therapy, and stage of contracture. PMID- 18391903 TI - Cemented primary total arthroplasty for acetabular protrusion in patients with rheumatoid arthritis. AB - BACKGROUND: The degenerative changes in hip joints observed in patients RA are associated with acetabular bone deficiencies. Therefore total hip arthroplasty performed in such patients requires bone grafting to restore the deficiencies. The aim of the study was to assess the long term-term results of primary total hip arthroplasty in patients with RA including an assessment of the usefulness of auto-and allograft in restoring bone deficiencies in acetabular protrusion. MATERIAL AND METHODS: From 1991 to 2001 a total of 156?? [w tekscie jest mowa o 158] patients with rheumatoid arthritis of the hip joint underwent a primary total arthroplasty of the hip joint with acetabular protrusion. The reconstruction of the acetabular bone was achieved by morsellized impaction auto and allografting. Group I included 26 patients who received autografts and Group II consisted of 10 patients who received allografts. The mean follow-up was 5 years. The hip joint performance was assessed according to the following scales: WOMAC, HHS, and Merle-d'Aubigne-Postel. Graft incorporation was graded according to the Avci criteria. RESULTS: The study revealed that auto-and allografting provides good long-term results. Graft incorporation was reported after 26 months. The radiographs showed porotic bone in grafts in one of the patients; however, there were no signs of loosening in the prosthetic acetabulum. CONCLUSION: Restoration of bone deficiencies in the medial part of the acetabulum in patients with RA by bone grafting is a biological way of reconstruction of the bone and facilitates stable fixation of a polyethylene acetabulum. PMID- 18391904 TI - The results of locked intramedullary nailing for non-union of forearm bones. AB - BACKGROUND: We report our experience with the use of a full-thickness corticocancellous iliac-crest graft, 1 to 5 centimeters in length - to treat established nonunion of the radius and/or ulna. MATERIAL AND METHODS: The technique was used in 14 patients (11 men and 3 women, age 19 to 59). The graft application was combined by rigid fixation with intramedullary locking nail, permitting early active mobilisation. We treated also one patient with infection complication of nonunion radius and ulna. In this case evacuation of metal plate, sequestrectomy, fixation external then antibiotics have been applied in first stage. After curing bone infection the grafting and fixation with the use of intramedullary locking nail have been made in second stage. RESULTS. The bone union has been achieved between 26-th and 33-rd week after operation in seven patients, in four patients with nonunion of radius and ulna bone union has been achieved after 33 weeks. In another two patients with graft 5 cm in length - bone union has been achieved after 28 weeks. In one patient with bone infection the grafts were good incorporated after 40 weeks. The motion of elbow and wrist in all cases has been satisfactory. CONCLUSIONS: Intramedullary nailing in treatment for forearm bones nonunion with use corticocancellous graft is technically easy, didn't require immobilization in the cast and enable early postoperative rehabilitation. PMID- 18391905 TI - Fixation of patellar tendon bone graft in reconstruction of patellar ligaments. Comparison of bioabsorbable and metal interference screws--results of treatment. AB - BACKGROUND: The central third of the patella tendon is probably the most common ACL graft material because the bone tendon bone (BTB) construct provides robust graft incorporation and a mechanically sufficient substitute. Metal and bioabsorbable interference screws ensure good initial fixation strength of the bone plug. MATERIAL AND METHODS: A total of 78 patients (mean age 29 years) were assigned to a bioabsorbable screw [L-15 Lactosorb] group (Group I of 34 patients) or a metal screw group (Group II of 44 patients). Mean follow-up was 3 years. This study compared the results of ACL reconstruction using bioabsorbable and metal interference screws. Pre- and postoperative assessments included patient history, the Lysholm Knee Scoring Scale and IKDC scores. Range of motion in the knee was measured. Magnetic resonance imaging was performed in 12 patients from each group. RESULTS: Mean Lysholm scores at one year were 94.2 for Group I and 93.5 for Group II. After 3 years mean scores were 95.4 for Group I and 96.2 for Group II. A comparison of IKDC scores showed no statistically significant difference between groups. No radiographic evidence of osteolytic change or bone resorption around the metal screw was observed in Group II, whereas osteolytic changes around bioscrews were observed in 2 patients in Group I. There were no complications related to loss of fixation. The clinical signs of a foreign body reaction were found in one patient. CONCLUSIONS: Bioabsorbale and metal interference screws produce equal clinical results. Complete degradation of a bioabsorbable screw takes 2-3 years. PMID- 18391906 TI - Bone mineral metabolism in children with juvenile idiopathic arthritis- preliminary report. AB - INTRODUCTION: There are very few reports assessing bone mineral mass and its metabolism in the course of juvenile idiopathic arthritis (JIA). STUDY OBJECTIVE: To assess the levels of selected serum markers of bone formation (OCN) and resorption (CTx) in JIA children. MATERIAL AND METHODS: The study involved 52 children with JIA diagnosed according to the EULAR criteria of 1997, aged 6-18 years. All patients underwent densitometric measurements using dual-energy X-ray absorptiometry (DXA) to assess TBBMD (g/cm2), Spine BMD (g/cm2), Z-score for SBMD, TBBMC (g), and LBM (g). The following parameters were determined in blood serum: the level of osteocalcin (OCN) and C-terminal type I alpha-collagen chain telopeptide (CTx) using the Elecsys 2010 system (N-MID Osteocalcin, Beta CrossLaps). A gender- and age-matched control group consisted of 16 healthy children. RESULTS: The mean concentrations of both osteocalcin (p<0.001) and CTx (p<0.005) were significantly higher in JIA patients as compared to the healthy controls (OCN 113.2+/-54.9 ng/ml vs. 70.2+/-48.3 ng/ml; CTx 1.4+/-0.5 mug/l vs. 1.2 +/-0.45 microg/l). The concentrations of the bone turnover markers were significantly reduced in children with higher degrees of joint destruction compared to those with anatomically normal joints (p<0.05). The mean concentration of CTx showed a significant negative correlation with the TB BMC/LBM Z-score (p<0.05). Reduced bone mass (Z-score for SBMD< -2.0) was found in 23.6% of the affected children. CONCLUSIONS: The JIA patients had elevated levels of OCN and CTx compared to the healthy controls. Reduced bone turnover was observed in children with higher degrees of joint destruction. PMID- 18391907 TI - The activity of fine afferent nerve fibres of the rat knee joint and their modulation by inflammatory mediators. AB - BACKGROUND: The origin of joint pain involves the activation of terminals of slowly conducting C and A-delta afferent fibres. The aim of this study was to characterize the slowly conducting nerve fibres supplying the rat knee joint and to illustrate the usefulness of this model for objective studies of the pathophysiological aspects of articular nociception and pain. MATERIAL AND METHODS: Using an in vivo model, single afferent fibres innervating normal and inflamed knee joints were isolated and electrophysiologically characterized. Responses of these fibres were examined after local mechanical stimulation (von Frey hairs) and rotations consisting of inward and outward rotations of the knee joint within (non-noxious) and outside (noxious stimuli) its normal working range. The chemosensitivity of afferent fibres was tested by applying excitatory and sensitizing agents. RESULTS: The nerve fibres supplying the rat knee joint responded to mechanical and chemical stimuli (bradykinin, capsaicin). Bradykinin, substance P and prostaglandin E2 sensitized a considerable percentage of nerve fibres to mechanical stimuli. CONCLUSIONS: The rat knee joint is a useful model to study nociception and inflammatory processes in an objective way. It can also be successfully used to study aspects of pain modulation. PMID- 18391908 TI - Operative treatment (subtotal resection) of cavernous hemangioma of forearm. A case report. AB - We present a case of cavernous hemangioma of a rare location within forearm muscles. The diagnosis was established on the basis of Magnetic Resonance Imaging and histological study of the specimen taken intraoperatively. A subtotal excision of the tumour was performed, followed by muscle reconstruction so that the hand function was preserved. After a follow-up period of five years, there were no clinical or MRI signs of recurrence. PMID- 18391911 TI - Advances in the treatment of myelodysplastic syndromes. AB - A Review of Selected Presentations From the 49th American Society of Hematology Annual Meeting and Exposition, December 8-11, 2007, Atlanta, Georgia. PMID- 18391909 TI - Problems with diagnostic work-up of musculoskeletal pain due to glomus tumour. Four case reports. AB - INTRODUCTION: Glomus tumours are rare benign neoplasms of myoepithelial origin. Due to characteristic and painful symptoms, glomus tumours are a major clinical problem. MATERIAL: Four histologically confirmed cases of glomangioma treated at the Orthopaedics & Traumatology Department between 1999 and 2007. RESULTS: The most significant symptom was extreme pain causing a decrease in the quality of life. Regardless of the location of the tumour, the correct diagnosis was delayed by 1.5 to 3 years. CONCLUSIONS: Due to its rare occurrence, glomus tumours are often missed out in the differential diagnosis of musculoskeletal pain. The agonising pain makes glomus tumours a significant clinical problem that should be remembered about, especially as surgery brings about spectacular improvement. PMID- 18391913 TI - Susceptibility to therapy-related myelodysplastic syndromes and acute myeloid leukemia. PMID- 18391914 TI - Combining radiotherapy with systemic therapies. PMID- 18391915 TI - K-Ras and sensitivity to EGFR inhibitors in metastatic colorectal cancer. PMID- 18391917 TI - Advances in the treatment of hemophilia. PMID- 18391918 TI - Erlotinib-induced acute hepatitis in a patient with pancreatic cancer. PMID- 18391919 TI - Review: hepatotoxicity and EGFR inhibition. PMID- 18391920 TI - Targeted therapies for hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) remains a highly lethal disease that is resistant to traditional cytotoxic chemotherapy. The last 30 years of chemotherapy clinical trials for advanced HCC have repeatedly failed to demonstrate any survival benefit for a long list of drugs. However a survival advantage was recently established for sorafenib, instituting a new standard of care for unresectable HCC. Here we review recent and ongoing studies of new therapeutic agents for HCC, including the small-molecule tyrosine kinase inhibitors, monoclonal antibodies, and combinations of these drugs. PMID- 18391922 TI - A metareview at the NIH. AB - Science funding in the United States is tight, and the application process is arduous. A recent study of NIH peer-review recommends a major overhaul of the system. Will the changes prove cosmetic or curative? PMID- 18391921 TI - Schwannoma of the kidney. AB - Schwannomas of the kidney are rare, with only a few reported cases. We report three additional cases with immunohistochemical analysis. All three tumors were from females (aged 27, 35, and 59 years) and ranged from 4.8 to 8 cm in diameter. All of the patients underwent nephrectomy. The tumors were totally or partially encapsulated; two were in the hilum and one was centered in the renal cortex. All tumors were diffusely positive for S100 protein. Two were positive for neuron specific enolase. Immunostaining for neurofilament, HMB45, microphthalmia transcription factor, smooth muscle actin, CD34, cytokeratin AE1/3, cytokeratin 7, and CD10 were negative. Follow-up data were available for two patients; neither had tumor recurrence or metastasis. In conclusion, renal schwannoma is rare, usually arises centrally, impinging on the hilum or the pelvis, and is cured by resection. Sarcomatoid carcinoma and other spindle cell tumors should be considered in the differential diagnosis. PMID- 18391923 TI - Search for potential autism treatments turns to 'trust hormone'. PMID- 18391924 TI - Gates Foundation looks to fund unconventional health solutions. PMID- 18391925 TI - After microbicide failures, hope that antiviral approach will gel. PMID- 18391926 TI - Gut instincts drive next generation of diabetes medications. PMID- 18391928 TI - Immune to the high. PMID- 18391929 TI - Q & A: Samir Khleif. AB - In 1999, Hussein bin Talal, the king of Jordan, died after a long battle with non Hodgkin's lymphoma. Since then, his eldest son and successor, King Abdullah II bin Al Hussein, has sought to improve cancer treatment in his country as part of an effort to boost healthcare and technological development. The country's capital, Amman, now boasts a world-class cancer treatment center, which draws patients from throughout the region. To lead that effort, the king tapped Samir Khleif, chief of the cancer vaccine section at the US National Cancer Institute in Bethesda, Maryland. Khleif, who received his medical degree in Jordan, now has an even bigger mandate from the king: to build an internationally renowned institute devoted to cancer research and biotechnology. Construction on The King Hussein Institute for Cancer and Biotechnology is scheduled to begin on the outskirts of Amman this month, with an expected completion in 2010. PMID- 18391931 TI - New drugs for an ancient parasite. PMID- 18391932 TI - Dropping acid to help cystic fibrosis. PMID- 18391933 TI - Getting personal about treating HIV. PMID- 18391934 TI - Bacteria fight back against Toll-like receptors. PMID- 18391935 TI - Robo4 counteracts VEGF signaling. PMID- 18391936 TI - Cancer's source in the peripheral nervous system. PMID- 18391938 TI - Brachytherapy for advanced prostate cancer bleeding. AB - Advanced prostate cancer patients frequently deal with intractable prostatic bleeding which is a difficult problem to manage. Intraurethral high-dose rate (HDR) brachytherapy may palliate this condition. Advanced prostate cancer patients with intractable prostatic bleeding were offered brachytherapy with Iridium-192 using a Micro-selectron HDR machine. During a 5-year period, analysis was performed in 23 patients with a median age and Gleason score of 78 years and 9, respectively. Following brachytherapy, haematuria resolved in 19 of the 23 patients and was recurrence free at 6 months. Intraurethral HDR brachytherapy is a potentially effective modality for treating haematuria in patients with advanced prostate cancer. PMID- 18391939 TI - Acute bacterial meningitis after transrectal needle biopsy of the prostate: a case report. AB - Transrectal biopsy of the prostate (TBP) is the most accurate method of diagnosing prostate cancer. Although debate remains, prophylactic antibiotic therapy is most frequently used to prevent infectious complications after TBP. Here, we present a case of Escherichia coli meningitis after TBP despite quinolone prophylaxis. PMID- 18391940 TI - p53 regulates glucose metabolism through an IKK-NF-kappaB pathway and inhibits cell transformation. AB - Cancer cells use aerobic glycolysis preferentially for energy provision and this metabolic change is important for tumour growth. Here, we have found a link between the tumour suppressor p53, the transcription factor NF-kappaB and glycolysis. In p53-deficient primary cultured cells, kinase activities of IKKalpha and IKKbeta and subsequent NF-kappaB activity were enhanced. Activation of NF-kappaB, by loss of p53, caused an increase in the rate of aerobic glycolysis and upregulation of Glut3. Oncogenic Ras-induced cell transformation and acceleration of aerobic glycolysis in p53-deficient cells were suppressed in the absence of p65/NF-kappaB expression, and were restored by GLUT3 expression. It was also shown that a glycolytic inhibitor diminished the enhanced IKK activity in p53-deficient cells. Moreover, in Ras-expressing p53-deficient cells, IKK activity was suppressed by p65 deficiency and restored by GLUT3 expression. Taken together, these data indicate that p53 restricts activation of the IKK-NF kappaB pathway through suppression of glycolysis. These results suggest that a positive-feedback loop exists, whereby glycolysis drives IKK-NF-kappaB activation, and that hyperactivation of this loop by loss of p53 is important in oncogene-induced cell transformation. PMID- 18391941 TI - Mature ribosomes are selectively degraded upon starvation by an autophagy pathway requiring the Ubp3p/Bre5p ubiquitin protease. AB - Eukaryotic cells use autophagy and the ubiquitin-proteasome system (UPS) as their major protein degradation pathways. Whereas the UPS is required for the rapid degradation of proteins when fast adaptation is needed, autophagy pathways selectively remove protein aggregates and damaged or excess organelles. However, little is known about the targets and mechanisms that provide specificity to this process. Here we show that mature ribosomes are rapidly degraded by autophagy upon nutrient starvation in Saccharomyces cerevisiae. Surprisingly, this degradation not only occurs by a non-selective mechanism, but also involves a novel type of selective autophagy, which we term 'ribophagy'. A genetic screen revealed that selective degradation of ribosomes requires catalytic activity of the Ubp3p/Bre5p ubiquitin protease. Although ubp3Delta and bre5Delta cells strongly accumulate 60S ribosomal particles upon starvation, they are proficient in starvation sensing and in general trafficking and autophagy pathways. Moreover, ubiquitination of several ribosomal subunits and/or ribosome-associated proteins was specifically enriched in ubp3Delta cells, suggesting that the regulation of ribophagy by ubiquitination may be direct. Interestingly, ubp3Delta cells are sensitive to rapamycin and nutrient starvation, implying that selective degradation of ribosomes is functionally important in vivo. Taken together, our results suggest a link between ubiquitination and the regulated degradation of mature ribosomes by autophagy. PMID- 18391942 TI - Corollary discharge circuits for saccadic modulation of the pigeon visual system. AB - A saccadic eye movement causes a variety of transient perceptual sequelae that might be the results of corollary discharge. Here we describe the neural circuits for saccadic corollary discharge that modulates activity throughout the pigeon visual system. Saccades in pigeons caused inhibition that was mediated by corollary discharge followed by enhancement of firing activity in the telencephalic hyperpallium, visual thalamus and pretectal nucleus lentiformis mesencephali (nLM) with opposite responses in the accessory optic nucleus (nBOR). Inactivation of thalamic neurons eliminated saccadic responses in telencephalic neurons, and inactivation of both the nLM and the nBOR abolished saccadic responses in thalamic neurons. Saccade-related omnipause neurons in the brainstem raphe complex inhibited the nBOR and excited the nLM, whereas inactivation of raphe neurons eliminated saccadic responses in both optokinetic and thalamic neurons. It seems that saccadic responses in telencephalic neurons are generated by corollary discharge signals from brainstem neurons that are transmitted through optokinetic and thalamic neurons. These signals might have important roles in visual perception. PMID- 18391943 TI - Silencing preBotzinger complex somatostatin-expressing neurons induces persistent apnea in awake rat. AB - Delineating neurons that underlie complex behaviors is of fundamental interest. Using adeno-associated virus 2, we expressed the Drosophila allatostatin receptor in somatostatin (Sst)-expressing neurons in the preBotzinger Complex (preBotC). Rapid silencing of these neurons in awake rats induced a persistent apnea without any respiratory movements to rescue their breathing. We hypothesize that breathing requires preBotC Sst neurons and that their sudden depression can lead to serious, even fatal, respiratory failure. PMID- 18391945 TI - TRPV1 shows dynamic ionic selectivity during agonist stimulation. AB - Transient receptor potential vanilloid 1 (TRPV1) is an ion channel that is gated by noxious heat, capsaicin and other diverse stimuli. It is a nonselective cation channel that prefers Ca2+ over Na+. These permeability characteristics, as in most channels, are widely presumed to be static. On the contrary, we found that activation of native or recombinant rat TRPV1 leads to time- and agonist concentration-dependent increases in relative permeability to large cations and changes in Ca2+ permeability. Using the substituted cysteine accessibility method, we saw that these changes were attributable to alterations in the TRPV1 selectivity filter. TRPV1 agonists showed different capabilities for evoking ionic selectivity changes. Furthermore, protein kinase C-dependent phosphorylation of Ser800 in the TRPV1 C terminus potentiated agonist-evoked ionic selectivity changes. Thus, the qualitative signaling properties of TRPV1 are dynamically modulated during channel activation, a process that probably shapes TRPV1 participation in pain, cytotoxicity and neurotransmitter release. PMID- 18391944 TI - Functional identification of sensory mechanisms required for developmental song learning. AB - A young male zebra finch (Taeniopygia guttata) learns to sing by copying the vocalizations of an older tutor in a process that parallels human speech acquisition. Brain pathways that control song production are well defined, but little is known about the sites and mechanisms of tutor song memorization. Here we test the hypothesis that molecular signaling in a sensory brain area outside of the song system is required for developmental song learning. Using controlled tutoring and a pharmacological inhibitor, we transiently suppressed the extracellular signal-regulated kinase signaling pathway in a portion of the auditory forebrain specifically during tutor song exposure. On maturation, treated birds produced poor copies of tutor song, whereas controls copied the tutor song effectively. Thus the foundation of normal song learning, the formation of a sensory memory of tutor song, requires a conserved molecular pathway in a brain area that is distinct from the circuit for song motor control. PMID- 18391947 TI - Single-molecule analysis of human telomerase monomer. AB - Human telomerase is a ribonucleoprotein that is minimally comprised of protein (hTERT) and RNA (hTR) components. We have applied single-molecule fluorescence two-color coincidence detection to characterize complex formation between fluorophore-labeled components in solution. By systematic labeling and in vitro assembly of hTERT, hTR and telomerase's DNA substrate, we have established that catalytically functional human telomerase comprises a stable hTERT:hTR:substrate interaction in a 1:1:1 absolute stoichiometry. PMID- 18391946 TI - Oligomeric amyloid-beta peptide disrupts phosphatidylinositol-4,5-bisphosphate metabolism. AB - Synaptic dysfunction caused by oligomeric assemblies of amyloid-beta peptide (Abeta) has been linked to cognitive deficits in Alzheimer's disease. Here we found that incubation of primary cortical neurons with oligomeric Abeta decreases the level of phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2), a phospholipid that regulates key aspects of neuronal function. The destabilizing effect of Abeta on PtdIns(4,5)P2 metabolism was Ca2+-dependent and was not observed in neurons that were derived from mice that are haploinsufficient for Synj1. This gene encodes synaptojanin 1, the main PtdIns(4,5)P2 phosphatase in the brain and at the synapses. We also found that the inhibitory effect of Abeta on hippocampal long-term potentiation was strongly suppressed in slices from Synj1+/- mice, suggesting that Abeta-induced synaptic dysfunction can be ameliorated by treatments that maintain the normal PtdIns(4,5)P2 balance in the brain. PMID- 18391948 TI - Highly active and selective endopeptidases with programmed substrate specificities. AB - A family of engineered endopeptidases has been created that is capable of cleaving a diverse array of peptide sequences with high selectivity and catalytic efficiency (kcat/KM > 10(40 M(- 1) s(- 1)). By screening libraries with a selection-counterselection substrate method, protease variants were programmed to recognize amino acids having altered charge, size and hydrophobicity properties adjacent to the scissile bond of the substrate, including GluArg, a specificity that to our knowledge has not been observed among natural proteases. Members of this artificial protease family resulted from a relatively small number of amino acid substitutions that (at least in one case) proved to be epistatic. PMID- 18391949 TI - Novel targets for Huntington's disease in an mTOR-independent autophagy pathway. AB - Autophagy is a major clearance route for intracellular aggregate-prone proteins causing diseases such as Huntington's disease. Autophagy induction with the mTOR inhibitor rapamycin accelerates clearance of these toxic substrates. As rapamycin has nontrivial side effects, we screened FDA-approved drugs to identify new autophagy-inducing pathways. We found that L-type Ca2+ channel antagonists, the K+ATP channel opener minoxidil, and the G(i) signaling activator clonidine induce autophagy. These drugs revealed a cyclical mTOR-independent pathway regulating autophagy, in which cAMP regulates IP3 levels, influencing calpain activity, which completes the cycle by cleaving and activating G(s)alpha, which regulates cAMP levels. This pathway has numerous potential points where autophagy can be induced, and we provide proof of principle for therapeutic relevance in Huntington's disease using mammalian cell, fly and zebrafish models. Our data also suggest that insults that elevate intracytosolic Ca2+ (like excitotoxicity) inhibit autophagy, thus retarding clearance of aggregate-prone proteins. PMID- 18391950 TI - Identification of ten loci associated with height highlights new biological pathways in human growth. AB - Height is a classic polygenic trait, reflecting the combined influence of multiple as-yet-undiscovered genetic factors. We carried out a meta-analysis of genome-wide association study data of height from 15,821 individuals at 2.2 million SNPs, and followed up the strongest findings in >10,000 subjects. Ten newly identified and two previously reported loci were strongly associated with variation in height (P values from 4 x 10(-7) to 8 x 10(-22)). Together, these 12 loci account for approximately 2% of the population variation in height. Individuals with < or =8 height-increasing alleles and > or =16 height-increasing alleles differ in height by approximately 3.5 cm. The newly identified loci, along with several additional loci with strongly suggestive associations, encompass both strong biological candidates and unexpected genes, and highlight several pathways (let-7 targets, chromatin remodeling proteins and Hedgehog signaling) as important regulators of human stature. These results expand the picture of the biological regulation of human height and of the genetic architecture of this classical complex trait. PMID- 18391951 TI - Many sequence variants affecting diversity of adult human height. AB - Adult human height is one of the classical complex human traits. We searched for sequence variants that affect height by scanning the genomes of 25,174 Icelanders, 2,876 Dutch, 1,770 European Americans and 1,148 African Americans. We then combined these results with previously published results from the Diabetes Genetics Initiative on 3,024 Scandinavians and tested a selected subset of SNPs in 5,517 Danes. We identified 27 regions of the genome with one or more sequence variants showing significant association with height. The estimated effects per allele of these variants ranged between 0.3 and 0.6 cm and, taken together, they explain around 3.7% of the population variation in height. The genes neighboring the identified loci cluster in biological processes related to skeletal development and mitosis. Association to three previously reported loci are replicated in our analyses, and the strongest association was with SNPs in the ZBTB38 gene. PMID- 18391952 TI - Genome-wide association analysis identifies 20 loci that influence adult height. AB - Adult height is a model polygenic trait, but there has been limited success in identifying the genes underlying its normal variation. To identify genetic variants influencing adult human height, we used genome-wide association data from 13,665 individuals and genotyped 39 variants in an additional 16,482 samples. We identified 20 variants associated with adult height (P < 5 x 10(-7), with 10 reaching P < 1 x 10(-10)). Combined, the 20 SNPs explain approximately 3% of height variation, with a approximately 5 cm difference between the 6.2% of people with 17 or fewer 'tall' alleles compared to the 5.5% with 27 or more 'tall' alleles. The loci we identified implicate genes in Hedgehog signaling (IHH, HHIP, PTCH1), extracellular matrix (EFEMP1, ADAMTSL3, ACAN) and cancer (CDK6, HMGA2, DLEU7) pathways, and provide new insights into human growth and developmental processes. Finally, our results provide insights into the genetic architecture of a classic quantitative trait. PMID- 18391953 TI - Rare independent mutations in renal salt handling genes contribute to blood pressure variation. AB - The effects of alleles in many genes are believed to contribute to common complex diseases such as hypertension. Whether risk alleles comprise a small number of common variants or many rare independent mutations at trait loci is largely unknown. We screened members of the Framingham Heart Study (FHS) for variation in three genes-SLC12A3 (NCCT), SLC12A1 (NKCC2) and KCNJ1 (ROMK)-causing rare recessive diseases featuring large reductions in blood pressure. Using comparative genomics, genetics and biochemistry, we identified subjects with mutations proven or inferred to be functional. These mutations, all heterozygous and rare, produce clinically significant blood pressure reduction and protect from development of hypertension. Our findings implicate many rare alleles that alter renal salt handling in blood pressure variation in the general population, and identify alleles with health benefit that are nonetheless under purifying selection. These findings have implications for the genetic architecture of hypertension and other common complex traits. PMID- 18391954 TI - Phosphatase SHP-1 promotes TLR- and RIG-I-activated production of type I interferon by inhibiting the kinase IRAK1. AB - Unbalanced production of proinflammatory cytokines and type I interferons in immune responses may lead to immunopathology; thus, the mechanisms that ensure the beneficial production of proinflammatory cytokines and type I interferons are of particular importance. Here we demonstrate that the phosphatase SHP-1 negatively regulated Toll-like receptor-mediated production of proinflammatory cytokines by inhibiting activation of the transcription factor NF-kappaB and mitogen-activated protein kinase. Simultaneously, SHP-1 increased the production of type I interferon mediated by Toll-like receptors and the helicase RIG-I by directly binding to and inhibiting activation of the kinase IRAK1. Our data demonstrate that SHP-1 contributes to immune homeostasis by balancing the production of proinflammatory cytokines and type I interferons in the innate immune response. PMID- 18391956 TI - Epitaxial graphene on ruthenium. AB - Graphene has been used to explore the fascinating electronic properties of ideal two-dimensional carbon, and shows great promise for quantum device architectures. The primary method for isolating graphene, micromechanical cleavage of graphite, is difficult to scale up for applications. Epitaxial growth is an attractive alternative, but achieving large graphene domains with uniform thickness remains a challenge, and substrate bonding may strongly affect the electronic properties of epitaxial graphene layers. Here, we show that epitaxy on Ru(0001) produces arrays of macroscopic single-crystalline graphene domains in a controlled, layer by-layer fashion. Whereas the first graphene layer indeed interacts strongly with the metal substrate, the second layer is almost completely detached, shows weak electronic coupling to the metal, and hence retains the inherent electronic structure of graphene. Our findings demonstrate a route towards rational graphene synthesis on transition-metal templates for applications in electronics, sensing or catalysis. PMID- 18391955 TI - Phosphatidylinositol-3-OH kinase and nutrient-sensing mTOR pathways control T lymphocyte trafficking. AB - Phosphatidylinositol-3-OH kinase (PI(3)K) and the nutrient sensor mTOR are evolutionarily conserved regulators of cell metabolism. Here we show that PI(3)K and mTOR determined the repertoire of adhesion and chemokine receptors expressed by T lymphocytes. The key lymph node-homing receptors CD62L (L-selectin) and CCR7 were highly expressed on naive T lymphocytes but were downregulated after immune activation. CD62L downregulation occurred through ectodomain proteolysis and suppression of gene transcription. The p110delta subunit of PI(3)K controlled CD62L proteolysis through mitogen-activated protein kinases, whereas control of CD62L transcription by p110delta was mediated by mTOR through regulation of the transcription factor KLF2. PI(3)K-mTOR nutrient-sensing pathways also determined expression of the chemokine receptor CCR7 and regulated lymphocyte trafficking in vivo. Hence, lymphocytes use PI(3)K and mTOR to match metabolism and trafficking. PMID- 18391957 TI - A chemically driven insulator-metal transition in non-stoichiometric and amorphous gallium oxide. AB - Insulator-metal transitions are well known in transition-metal oxides, but inducing an insulator-metal transition in the oxide of a main group element is a major challenge. Here, we report the observation of an insulator-metal transition, with a conductivity jump of seven orders of magnitude, in highly non stoichiometric, amorphous gallium oxide of approximate composition GaO(1.2) at a temperature around 670 K. We demonstrate through experimental studies and density functional-theory calculations that the conductivity jump takes place at a critical gallium concentration and is induced by crystallization of stoichiometric Ga(2)O(3) within the metastable oxide matrix-in chemical terms by a disproportionation. This novel mechanism--an insulator-metal transition driven by a heterogeneous solid-state reaction--opens up a new route to achieve metallic behaviour in oxides that are expected to exist only as classic insulators. PMID- 18391958 TI - Improved genetic manipulation of human embryonic stem cells. AB - Low efficiency of transfection limits the ability to genetically manipulate human embryonic stem cells (hESCs), and differences in cell derivation and culture methods require optimization of transfection protocols. We transiently transferred multiple independent hESC lines with different growth requirements to standardized feeder-free culture, and optimized conditions for clonal growth and efficient gene transfer without loss of pluripotency. Stably transfected lines retained differentiation potential, and most lines displayed normal karyotypes. PMID- 18391960 TI - Spheroid-based engineering of a human vasculature in mice. AB - The complexity of the angiogenic cascade limits cellular approaches to studying angiogenic endothelial cells (ECs). In turn, in vivo assays do not allow the analysis of the distinct cellular behavior of ECs during angiogenesis. Here we show that ECs can be grafted as spheroids into a matrix to give rise to a complex three-dimensional network of human neovessels in mice. The grafted vasculature matures and is connected to the mouse circulation. The assay is highly versatile and facilitates numerous applications including studies of the effects of different cytokines on angiogenesis. Modifications make it possible to study human lymphangiogenic processes in vivo. EC spheroids can also be coimplanted with other cell types for tissue engineering purposes. PMID- 18391959 TI - BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals. AB - The interpretation of genome sequences requires reliable and standardized methods to assess protein function at high throughput. Here we describe a fast and reliable pipeline to study protein function in mammalian cells based on protein tagging in bacterial artificial chromosomes (BACs). The large size of the BAC transgenes ensures the presence of most, if not all, regulatory elements and results in expression that closely matches that of the endogenous gene. We show that BAC transgenes can be rapidly and reliably generated using 96-well-format recombineering. After stable transfection of these transgenes into human tissue culture cells or mouse embryonic stem cells, the localization, protein-protein and/or protein-DNA interactions of the tagged protein are studied using generic, tag-based assays. The same high-throughput approach will be generally applicable to other model systems. PMID- 18391961 TI - Dopamine neurons implanted into people with Parkinson's disease survive without pathology for 14 years. AB - Postmortem analysis of five subjects with Parkinson's disease 9-14 years after transplantation of fetal midbrain cell suspensions revealed surviving grafts that included dopamine and serotonin neurons without pathology. These findings are important for the understanding of the etiopathogenesis of midbrain dopamine neuron degeneration and future use of cell replacement therapies. PMID- 18391962 TI - Lewy body-like pathology in long-term embryonic nigral transplants in Parkinson's disease. AB - Fourteen years after transplantation into the striatum of an individual with Parkinson's disease, grafted nigral neurons were found to have Lewy body-like inclusions that stained positively for alpha-synuclein and ubiquitin and to have reduced immunostaining for dopamine transporter. These pathological changes suggest that Parkinson's disease is an ongoing process that can affect grafted cells in the striatum in a manner similar to host dopamine neurons in the substantia nigra. These findings have implications for cell-based therapies and for understanding the cause of Parkinson's disease. PMID- 18391963 TI - Lewy bodies in grafted neurons in subjects with Parkinson's disease suggest host to-graft disease propagation. AB - Two subjects with Parkinson's disease who had long-term survival of transplanted fetal mesencephalic dopaminergic neurons (11-16 years) developed alpha-synuclein positive Lewy bodies in grafted neurons. Our observation has key implications for understanding Parkinson's pathogenesis by providing the first evidence, to our knowledge, that the disease can propagate from host to graft cells. However, available data suggest that the majority of grafted cells are functionally unimpaired after a decade, and recipients can still experience long-term symptomatic relief. PMID- 18391964 TI - Sequence-directed DNA export guides chromosome translocation during sporulation in Bacillus subtilis. AB - In prokaryotes, the transfer of DNA between cellular compartments is essential for the segregation and exchange of genetic material. SpoIIIE and FtsK are AAA+ ATPases responsible for intercompartmental chromosome translocation in bacteria. Despite functional and sequence similarities, these motors were proposed to use drastically different mechanisms: SpoIIIE was suggested to be a unidirectional DNA transporter that exports DNA from the compartment in which it assembles, whereas FtsK was shown to establish translocation directionality by interacting with highly skewed chromosomal sequences. Here we use a combination of single molecule, bioinformatics and in vivo fluorescence methodologies to study the properties of DNA translocation by SpoIIIE in vitro and in vivo. These data allow us to propose a sequence-directed DNA exporter model that reconciles previously proposed models for SpoIIIE and FtsK, constituting a unified model for directional DNA transport by the SpoIIIE/FtsK family of AAA+ ring ATPases. PMID- 18391965 TI - A conserved rRNA methyltransferase regulates ribosome biogenesis. AB - In contrast to the diversity of most ribosomal RNA modification patterns and systems, the KsgA methyltransferase family seems to be nearly universally conserved along with the modifications it catalyzes. Our data reveal that KsgA interacts with small ribosomal subunits near functional sites, including Initiation factor 3 and 50S subunit binding sites. These findings suggest a checkpoint role for this modification system and offer a functional rationale for the unprecedented level of conservation. PMID- 18391966 TI - Signal sequence-independent membrane targeting of ribosomes containing short nascent peptides within the exit tunnel. AB - Ribosomes synthesizing inner membrane proteins in Escherichia coli are targeted to the translocon in the plasma membrane by the signal recognition particle (SRP) and the SRP receptor, FtsY. Here we show using a purified system that membrane targeting does not require an exposed signal-anchor sequence, as SRP-dependent targeting takes place with ribosomes containing short nascent peptides, with or without a signal-anchor sequence, within the peptide exit tunnel. Signaling from inside the tunnel involves ribosomal protein L23, which constitutes part of the SRP binding site. When nascent peptides emerge from the ribosome, the targeting complex is maintained with ribosomes exposing a signal-anchor sequence, whereas ribosomes exposing other sequences are released. These results indicate that ribosome-nascent chain complexes containing any nascent peptide within the exit tunnel can enter the SRP targeting pathway to be sorted at the membrane into ribosome-nascent chain complexes that synthesize either membrane or cytosolic proteins. PMID- 18391968 TI - A brief introduction to FOXOlogy. AB - Members of the Forkhead box O (FOXO) class of transcription factors are key players in the regulation of cell-fate decisions, such as cell death, cell proliferation and cell metabolism. Furthermore, in model organisms, it has by now been demonstrated that FOXO function affects the life span of these organisms. Multiple signal transduction pathways regulate FOXO function, but most importantly, they are negatively regulated by protein kinase B (PKB/AKT)-mediated phosphorylation and constitute, therefore, an important downstream component of insulin signalling. This review issue provides a timely overview of our understanding of FOXO function and how signalling affects FOXO function. Taken together, the reviewed studies on FOXO function and regulation provide compelling evidence that FOXOs act at the crossroad between aging and age-related diseases including diabetes and cancer. With this perspective, further studies on FOXO function and regulation may shed light on how age impacts on the onset and progression of disease. PMID- 18391969 TI - Structure/function relationships underlying regulation of FOXO transcription factors. AB - The FOXO subgroup of forkhead transcription factors plays a central role in cell cycle control, differentiation, metabolism control, stress response and apoptosis. Therefore, the function of these important molecules is tightly controlled by a wide range of protein-protein interactions and posttranslational modifications including phosphorylation, acetylation and ubiquitination. The mechanisms by which these processes regulate FOXO activity are mostly elusive. This review focuses on recent advances in structural studies of forkhead transcription factors and the insights they provide into the mechanism of DNA recognition. On the basis of these data, we discuss structural aspects of protein protein interactions and posttranslational modifications that target the forkhead domain and the nuclear localization signal of FOXO proteins. PMID- 18391970 TI - The FoxO code. AB - The FoxO family of Forkhead transcription factors plays an important role in longevity and tumor suppression by upregulating target genes involved in stress resistance, metabolism, cell cycle arrest and apoptosis. FoxO transcription factors translate a variety of environmental stimuli, including insulin, growth factors, nutrients and oxidative stress, into specific gene-expression programs. These environmental stimuli control FoxO activity primarily by regulating their subcellular localization, but also by affecting their protein levels, DNA-binding properties and transcriptional activity. The precise regulation of FoxO transcription factors is enacted by an intricate combination of post translational modifications (PTMs), including phosphorylation, acetylation and ubiquitination, and binding protein partners. An intriguing possibility is that FoxO PTMs may act as a 'molecular FoxO code' read by selective protein partners to rapidly regulate gene-expression programs. The effective control of FoxO activity in response to environmental stimuli is likely to be critical to prevent aging and age-dependent diseases, including cancer, neurodegenerative diseases and diabetes. PMID- 18391971 TI - FOXO-binding partners: it takes two to tango. AB - Modulation FOXO transcription factor activities can lead to a variety of cellular outputs resulting in changes in proliferation, apoptosis, differentiation and metabolic responses. Although FOXO proteins all contain an identical DNA-binding domain their cellular functions appear to be distinct, as exemplified by differences in the phenotype of Foxo1, Foxo3 and Foxo4 null mutant mice. While some of these differences may be attributable to the differential expression patterns of these transcription factors, many cells and tissues express several FOXO isoforms. Recently it has become clear that FOXO proteins can regulate transcriptional responses independently of direct DNA-binding. It has been demonstrated that FOXOs can associate with a variety of unrelated transcription factors, regulating activation or repression of diverse target genes. The complement of transcription factors expressed in a particular cell type is thus critical in determining the functional end point of FOXO activity. These interactions greatly expand the possibilities for FOXO-mediated regulation of transcriptional programmes. This review details currently described FOXO-binding partners and examines the role of these interactions in regulating cell fate decisions. PMID- 18391972 TI - Many forks in the path: cycling with FoxO. AB - FoxO transcription factors are an evolutionary conserved subfamily of the forkhead transcription factors, characterized by the forkhead DNA-binding domain. FoxO factors regulate a number of cellular processes involved in cell-fate decisions in a cell-type- and environment-specific manner, including metabolism, differentiation, apoptosis and proliferation. A key mechanism by which FoxO determines cell fate is through regulation of the cell cycle machinery, and as such the cellular consequence of FoxO deregulation is often manifested through perturbation of the cell cycle. Consequently, the deregulation of FoxO factors is implicated in the development of numerous proliferative diseases, in particular cancer. PMID- 18391973 TI - FOXOs, cancer and regulation of apoptosis. AB - Forkhead box O (FOXO) transcription factors are involved in multiple signaling pathways and play critical roles in a number of physiological and pathological processes including cancer. The importance of FOXO factors ascribes them under multiple levels of regulation including phosphorylation, acetylation/deacetylation, ubiquitination and protein-protein interactions. As FOXO factors play a pivotal role in cell fate decision, mounting evidence suggests that FOXO factors function as tumor suppressors in a variety of cancers. FOXOs are actively involved in promoting apoptosis in a mitochondria-independent and -dependent manner by inducing the expression of death receptor ligands, including Fas ligand and tumor necrosis factor-related apoptosis-inducing ligand, and Bcl-2 family members, such as Bim, bNIP3 and Bcl-X(L), respectively. An understanding of FOXO proteins and their biology will provide new opportunities for developing more effective therapeutic approaches to treat cancer. PMID- 18391974 TI - The role of FoxO in the regulation of metabolism. AB - Forkhead proteins, and FoxO1 in particular, play a significant role in regulating whole body energy metabolism. Glucose homeostasis is achieved by adjusting endogenous glucose production as well as glucose uptake by peripheral tissues in response to insulin. In the fasted state, the liver is primarily responsible for maintaining glucose levels, with FoxO1 playing a key role in promoting the expression of gluconeogenic enzymes. Following feeding, pancreatic beta cells secrete insulin, which promotes the uptake of glucose by peripheral tissues including skeletal muscle and adipose tissue, and can in part suppress gluconeogenic enzyme expression in the liver. In addition to directly regulating metabolism, FoxO1 also plays a role in the formation of both adipose tissue and skeletal muscle, two major organs that are critical for maintaining energy homeostasis. The importance of FoxO1 in energy homeostasis is particularly striking under conditions of metabolic dysfunction or insulin resistance. In obese or diabetic states, FoxO1-dependent gene expression promotes some of the deleterious characteristics associated with these conditions, including hyperglycemia and glucose intolerance. In addition, the increase in pancreatic beta cell mass that normally occurs in response to a rise in insulin demand is blunted by nuclear FoxO1 expression. However, under these same pathophysiological conditions, FoxO1 expression may help drive the expression of genes involved in combating oxidative stress, thereby preserving cellular function. FoxO1 may also be involved in promoting the switch from carbohydrate to fatty acid as the major energy source during starvation. PMID- 18391975 TI - Foxo in the immune system. AB - In addition to their key roles in cellular survival, death, proliferation and metabolism, the Foxo subfamily of forkhead (Fox) transcription factors play critical roles in the homeostasis of immune-relevant cells, including T cells, B cells, neutrophils and other non-lymphoid lineages that modulate inflammation in disease states such as inflammatory arthritis and systemic lupus erythematosus. This review summarizes such current and expanding knowledge of the Foxo family members in immunity, and their potential as therapeutic targets in inflammatory disease. PMID- 18391976 TI - FOXO animal models reveal a variety of diverse roles for FOXO transcription factors. AB - The Foxo subfamily of FOX transcription factors plays a variety of roles in a broad assortment of diverse physiological processes including cellular differentiation, tumor suppression, metabolism, cell cycle arrest, cell death and protection from stress. Animal models have proved to be invaluable tools in furthering our understanding of the role of particular genes in complex organismal processes. Multiple animal models in diverse species, including Caenorhabditis elegans, Drosophila. melanogaster and the laboratory mouse, exist for the Foxo family of transcription factors. Foxo genes are highly conserved throughout the evolution and each of these model systems has provided valuable insight into the roles of Foxo factors. Many roles are conserved among the different model organisms. Several Foxo-related animal model systems are reviewed here along with the knowledge gleaned to date from each model system. PMID- 18391977 TI - Forkhead transcription factors and ageing. AB - Mutations in single genes and environmental interventions can extend healthy lifespan in laboratory model organisms. Some of the mechanisms involved show evolutionary conservation, opening the way to using simpler invertebrates to understand human ageing. Forkhead transcription factors have been found to play a key role in lifespan extension by alterations in the insulin/IGF pathway and by dietary restriction. Interventions that extend lifespan have also been found to delay or ameliorate the impact of ageing-related pathology and disease, including cancer. Understanding the mode of action of forkheads in this context will illuminate the mechanisms by which ageing acts as a risk factor for ageing related disease, and could lead to the development of a broad-spectrum, preventative medicine for the diseases of ageing. PMID- 18391978 TI - DNA amplification is a ubiquitous mechanism of oncogene activation in lung and other cancers. AB - Chromosomal translocation is the best-characterized genetic mechanism for oncogene activation. However, there are documented examples of activation by alternate mechanisms, for example gene dosage increase, though its prevalence is unclear. Here, we answered the fundamental question of the contribution of DNA amplification as a molecular mechanism driving oncogenesis. Comparing 104 cancer lines representing diverse tissue origins identified genes residing in amplification 'hotspots' and discovered an unexpected frequency of genes activated by this mechanism. The 3431 amplicons identified represent approximately 10 per hematological and approximately 36 per epithelial cancer genome. Many recurrently amplified oncogenes were previously known to be activated only by disease-specific translocations. The 135 hotspots identified contain 538 unique genes and are enriched for proliferation, apoptosis and linage dependency genes, reflecting functions advantageous to tumor growth. Integrating gene dosage with expression data validated the downstream impact of the novel amplification events in both cell lines and clinical samples. For example, multiple downstream components of the EGFR-family-signaling pathway, including CDK5, AKT1 and SHC1, are overexpressed as a direct result of gene amplification in lung cancer. Our findings suggest that amplification is far more common a mechanism of oncogene activation than previously believed and that specific regions of the genome are hotspots of amplification. PMID- 18391979 TI - Loss of Rassf1a cooperates with Apc(Min) to accelerate intestinal tumourigenesis. AB - Promoter methylation of the RAS-association domain family 1, isoform A gene (RASSF1A) is one of the most frequent events found in human tumours. In this study we set out to test the hypothesis that loss of Rassf1a can cooperate with inactivation of the adenomatous polyposis coli (Apc) gene to accelerate intestinal tumourigenesis using the Apc-Min (Apc(Min/+)) mouse model, as mutational or deletional inactivation of APC is a frequent early event in the genesis of intestinal cancer. Further, loss of RASSF1A has also been reported to occur in premalignant adenomas of the bowel. RASSF1A has been implicated in an array of pivotal cellular processes, including regulation of the cell cycle, apoptosis, microtubule stability and most recently in the beta-catenin signalling pathway. By interbreeding isoform specific Rassf1a knockout mice with Apc(+/Min) mice, we showed that loss of Rassf1a results in a significant increase in adenomas of the small intestine and accelerated intestinal tumourigenesis leading to the earlier death of adenocarcinoma-bearing mice and decreased overall survival. Comparative genomic hybridization of adenomas from Rassf1a(-/-); Apc(+/Min) mice revealed no evidence of aneuploidy or gross chromosomal instability (no difference to adenomas from Rassf1a(+/+); Apc(+/Min) mice). Immunohistochemical analysis of adenomas revealed increased nuclear beta-catenin accumulation in adenomas from Rassf1a(-/-); Apc(+/Min) mice, compared to those from Rassf1a(+/+); Apc(+/Min) mice, but no differences in proliferation marker (Ki67) staining patterns. Collectively these data demonstrate cooperation between inactivation of Rassf1a and Apc resulting in accelerated intestinal tumourigenesis, with adenomas showing increased nuclear accumulation of beta catenin, supporting a mechanistic link via loss of the known interaction of Rassf1 with beta-TrCP that usually mediates degradation of beta-catenin. PMID- 18391980 TI - Functional inactivation of the WTX gene is not a frequent event in Wilms' tumors. AB - For many years the precise genetic etiology of the majority of Wilms' tumors has remained unexplained. Recently, the WTX gene, mapped to chromosome Xq11.1, has been reported to be lost or mutated in approximately one-third of Wilms' tumors. Moreover, in female cases, the somatically inactivated alleles were found to invariantly derive from the active chromosome X. Consequently, WTX has been proposed as a 'one-hit' tumor suppressor gene. To provide further insights on the contribution of WTX to the development of the disease, we have examined 102 Wilms' tumors, obtained from 43 male and 57 female patients. Quantitative PCR analyses detected WTX deletions in 5 of 45 (11%) tumors from males, whereas loss of heterozygosity at WTX-linked microsatellites was observed in 9 tumors from 50 informative females (19%). However, in the latter group, using a combination of HUMARA assay and bisulfite-modified DNA sequencing, we found that the deletion affected the active chromosome X only in two cases (4%). Sequence analyses detected an inactivating somatic mutation of WTX in a single tumor, in which a strongly reduced expression of the mutant allele respect to the wild-type allele was observed, a finding not consistent with its localization on the active chromosome X. Overall, a functional somatic nullizygosity of the WTX gene was ascertained only in seven of the Wilms' tumors included in the study (approximately 7%). Our findings indicate that previously reported estimates on the proportion of Wilms' tumors due to WTX alterations should be reconsidered. PMID- 18391981 TI - AIP regulates stability of Aurora-A at early mitotic phase coordinately with GSK 3beta. AB - Glycogen synthase kinase-3 (GSK-3beta) regulates microtubule dynamics and cellular polarity through phosphorylating various microtubule associating proteins and plus-end tracking proteins. Although it was also reported that GSK 3beta is inactivated by protein kinase B at the spindle poles, functions and targets of GSK-3beta in the mitotic phase are unknown. Here, we identified Aurora A-interacting protein (AIP), a negative regulator of Aurora-A, as a binding partner of GSK-3beta. AIP was colocalized with Aurora-A and GSK-3beta to the spindle poles in metaphase, and its depletion in cells stabilized and activated Aurora-A in early mitotic phase and caused mitotic cell arrest. Treatment of the cells with a GSK-3beta inhibitor reduced the protein level of Aurora-A and this reduction was suppressed by AIP knockdown. AIP was phosphorylated by GSK-3beta, and an AIP mutant in which the GSK-3beta phosphorylation site was mutated could bind and downregulate Aurora-A more efficiently. These results suggest that GSK 3beta modulates the early mitotic Aurora-A level through binding and phosphorylating AIP. PMID- 18391982 TI - Cancer-specific enhancement of cisplatin-induced cytotoxicity with triptolide through an interaction of inactivated glycogen synthase kinase-3beta with p53. AB - To improve conventional chemotherapeutic efficacy, a combination use of traditional medicines is effective but detailed mechanisms have been rarely elucidated. In the this study, we attempted to clarify how triptolide (PG490), an oxygenated diterpene derived from a Chinese herb, enhances the cisplatin (CDDP) induced cytotoxicity in urothelial cancer cells. Our results showed that a combined CDDP/triptolide therapy induced apoptosis in urothelial cancer cell lines with wild-type p53, but not in those with mutant-type p53 or normal human urothelium. As the mechanism, triptolide suppressed CDDP-induced p53 transcriptional activity, leading to p21 attenuation, which promoted apoptosis via the activation of c-Jun N-terminal kinase (JNK) and Bax. We further demonstrated that the functional regulation of p53 by triptolide was mediated by an intranuclear association of p53 with glycogen synthase kinase-3beta (GSK3beta), which was inactivated by protein kinase C (PKC). This modulation of the PKC-GSK3beta axis by triptolide was observed in a cancer-specific manner. A mouse xenograft model also showed that a combined CDDP/triptolide therapy completely suppressed tumor growth without any side effects. We expect that cancer-specific enhancement of CDDP-induced cytotoxicity with triptolide may effectively overcome the resistance to a CDDP-based conventional chemotherapy as a treatment for urothelial cancer. PMID- 18391983 TI - Activated c-Abl tyrosine kinase in malignant solid tumors. AB - Mutant forms of the c-ABL gene are well known to be involved in hematopoietic malignancies such as chronic myeloid leukemia (CML). CML patients possess a fused BCR-ABL gene that activates the Abl tyrosine kinase domain within Bcr-Abl. In general fusion proteins that cause oligomerization of Abl lead to activation of its tyrosine kinase activity. In this review, we highlight recent discoveries indicating that the activated c-Abl tyrosine kinase, not as a fusion protein, plays an important role in malignant solid tumors of lung and breast. PMID- 18391984 TI - FOXO3a mediates the androgen-dependent regulation of FLIP and contributes to TRAIL-induced apoptosis of LNCaP cells. AB - Androgen-withdrawal-induced apoptosis (AWIA) is deregulated in androgen refractory prostate cancer. Androgens have been shown to positively regulate expression of the antiapoptotic FADD-like interleukin-1beta-converting enzyme (FLICE)-like inhibitory protein (FLIP), and reduced FLIP expression precedes apoptosis after androgen withdrawal. Here, we show that FLIP protein expression is downregulated in castrated rats, while in LNCaP cells, androgens regulate FLIP in a manner that is dependent on phosphoinositol-3-kinase (PI3K) and Akt signaling. Specifically, treatment of LNCaP cells with LY294002, or expression of either PTEN or a non-phosphorylatable form of FOXO3a (FOXO3aTM), downregulates FLIP protein and mRNA. Conversely, treatment with androgens in the absence of PI3/Akt signaling, or following expression of FOXO3aTM, leads to increased FLIP expression. A FOXO3a binding site was identified in the FLIP promoter and shown necessary for the combined effects of androgens and FOXO3a on FLIP transcription. FOXO3a binds the androgen receptor, suggesting that the transcriptional synergy depends on an interaction between these proteins. Finally, LNCaP cells are sensitized to TRAIL-induced apoptosis by PTEN or LY294002, and rescued by androgens. FOXO3aTM also sensitizes cells to androgen-inhibited TRAIL apoptosis. Androgen rescue was diminished when either FOXO3a or FLIP was reduced by siRNA. These data support a role for FOXO3a in AWIA. PMID- 18391985 TI - Fbw7 regulates the activity of endoreduplication mediators and the p53 pathway to prevent drug-induced polyploidy. AB - Fbw7 is a tumor suppressor that is mutated in numerous cancers. It encodes an E3 ubiquitin ligase, whose ability to decrease the levels of pivotal regulators of cell growth and proliferation underlies its tumor suppressor function. Here, we explore the consequences of Fbw7 inactivation on the outcome of chemotherapeutic treatments. When exposed to spindle toxins such as vinblastine and taxol, Fbw7 deficient cells undergo extensive mitotic slippage and endoreduplication, rendering them polyploid. A combined deregulation of several Fbw7 target proteins is required for this phenotype. Specifically, elevated expression of cyclin E and Aurora A in Fbw7-deficient cells is required for drug-induced polyploidy. However, overexpression of either cyclin E or Aurora A alone is not sufficient for drug-induced polyploidy. In addition, we demonstrate that Fbw7 deficiency limits the ability of p53 to respond to mitotic toxins but not to DNA damage. Furthermore, Fbw7 expression regulates the p53-dependent induction of genes such as Lats2 and p21 in response to vinblastine. Hence, we suggest that Fbw7 serves as a master regulator of the mitotic and tetraploidy checkpoints. PMID- 18391986 TI - Loss of p53 induces epidermal growth factor receptor promoter activity in normal human keratinocytes. AB - Overexpression of the epidermal growth factor receptor (EGFR) in human papillomavirus type 16-immortalized human keratinocytes (HKc) is caused by the viral oncoprotein E6, which targets p53 for degradation. We have previously observed that expression of p53 RNAi in normal HKc is associated with an increase in EGFR mRNA and protein. We now report that p53 RNAi induces EGFR promoter activity up to approximately 10-fold in normal HKc, and this effect does not require intact p53 binding sites on the EGFR promoter. Exogenous wild-type p53 inhibits the EGFR promoter at low levels, and activates it at higher concentrations. Yin Yang 1 (YY1), which negatively regulates p53, induces EGFR promoter activity, and this effect is augmented by p53 RNAi. Intact p53 binding sites on the EGFR promoter are not required for activation by YY1. In addition, Sp1 and YY1 synergistically induce the EGFR promoter in normal HKc, indicating that Sp1 may recruit YY1 as a co-activator. Wild-type p53 suppressed Sp1- and YY1 mediated induction of the EGFR promoter. We conclude that acute loss of p53 in normal HKc induces EGFR expression by a mechanism that involves YY1 and Sp1 and does not require p53 binding to the EGFR promoter. PMID- 18391987 TI - Allo-SCT using reduced-intensity conditioning against advanced pancreatic cancer: a Japanese survey. AB - Pancreatic cancer is a frequent cause of cancer-related mortality and has an extremely poor prognosis. To evaluate the efficacy of allogeneic hematopoietic SCT with reduced-intensity conditioning (RICT) against pancreatic cancer, we analyzed the clinical data of 22 patients. After a fludarabine-based conditioning regimen followed by the infusion of PBSCs, all but two achieved engraftment. Complete, partial and minor response was observed in 1, 2 and 2 patients, respectively, with an overall response rate of 23%. Median survival was only 139 days and the major cause of death was tumor progression. Poor performance status before RICT and a lower number of infused CD34-positive cells were associated with shorter survival after RICT. Patients who developed chronic GVHD tended to survive longer than those who did not. These findings support the investigation of a novel treatment strategy to enhance the immunological effect against pancreatic cancer. PMID- 18391988 TI - Transportation and cryopreservation may impair haematopoietic stem cell function and engraftment of allogeneic PBSCs, but not BM. AB - Recent data suggest that the practice of using frozen allogeneic grafts is becoming increasingly common among transplant centres. Therefore, we retrospectively analysed 31 frozen allogeneic PBSC and 8 BM grafts by flow cytometry with regard to their CD34+ content, membrane integrity (7-AAD) and stem cell-specific enzyme activity (aldehyde dehydrogenase, ALDH) in relation to individual transplantation results. Membrane integrity of CD34+ cells was significantly impaired in cryopreserved PBSC but not in BM compared to unfrozen allografts. In 9 out of 31 frozen PBSC (but none of the BM) grafts numbers of SSC(lo)ALDH(br) cells per kg body weight (BW) were significantly reduced while in the same grafts the numbers of CD34+ cells per kg BW were close to normal. Overall, 9 out of 33 patients (27%) who received unrelated PBSC allografts cryopreserved after transportation did not achieve engraftment. For comparison, primary graft failure was observed in our centre in only 7 out of 493 recipients (1.4%) of fresh allogeneic PBSC grafts. Moreover, we did not see any graft failure in patients receiving frozen/thawed BM or autologous PBSC transplants. We, therefore, conclude that PBSC grafts become much more sensitive to cryopreservation after transport and/or storage. Importantly, the engraftment potential of frozen HSC grafts may reliably be predicted by measuring ALDH activity. PMID- 18391989 TI - Impact of germinal center and non-germinal center phenotypes on overall and failure-free survival after high-dose chemotherapy and auto-SCT in primary diffuse large B-cell lymphoma. AB - Non-germinal center (non-GC) phenotype is an adverse prognostic factor in chemotherapy (CT)-treated diffuse large B-cell lymphoma (DLBCL) patients. To determine how high-dose therapy (HDT) supported with auto-SCT as first line therapy influences GC-associated outcome in young high-risk DLBCL patients GC and non-GC phenotypes were determined immunohistochemically from 63 patients. Of these, 29 primary high-risk DLBCL patients were treated with auto-SCT, whereas 34 CT-treated patients served as a control group. Consistent with previous studies, non-GC phenotype was associated with adverse outcome in CT-treated high-risk patients. In contrast, immunohistochemical classification by cell of origin did not associate with survival after auto-SCT. When the impact of treatment on the predictive value of cell of origin was analyzed, the non-GC patients, who received HDT, had a better failure-free survival (FFS) and overall survival (OS) than the patients treated with CT alone. In multivariate analyses, both age adjusted International Prognostic Index (aaIPI) and treatment were independent prognostic factors for FFS and OS. For the patients with GC phenotype, the influence of auto-SCT on survival was not significant. The data imply that auto SCT can overcome the adverse prognostic impact of the non-GC phenotype in patients with high-risk DLBCL and warrant additional prospective studies. PMID- 18391990 TI - Increasing mixed chimerism and the risk of graft loss in children undergoing allogeneic hematopoietic stem cell transplantation for non-malignant disorders. AB - We performed quantitative PCR-based serial chimerism testing of whole blood (WB) and CD3+ cells and retrospectively correlated the results of chimerism tests and the risk of graft loss in children undergoing transplant for non-malignant disorders. Twenty-four children were included in this study. All patients initially engrafted; subsequently, 12% lost the graft, 21% achieved complete donor chimerism and 67% had mixed chimerism (MC). Patients underwent delayed taper of cyclosporine (CsA) if they had MC. Overall survival was 87+/-7% (s.d.) at 5-years post transplant, and it was not affected by chimerism status. Both WB and CD3+ chimerism showed significant fluctuations with a peak in autologous cell signal occurring at a median of 7 months for WB and 2 months for CD3+ cells. Initial post transplant chimerism percentage in either WB or CD3+ lineage was not related to graft loss. Increasing MC to >30% host cells was seen in 33% of patients, and it was related to increased risk of graft loss, as previously published. However, 63% of children with increasing MC did not lose their graft. Additional studies of post transplant chimerism are required to improve our ability to accurately identify children at risk of graft loss following transplant for non-malignant disorders. PMID- 18391991 TI - Positron emission tomography using F-18 fluorodeoxyglucose pre- and post autologous stem cell transplant in non-Hodgkin's lymphoma. AB - Positron emission tomography (PET) utilizing fluorodeoxyglucose (FDG) has an ever increasing role in the management of numerous malignancies. FDG PET in lymphoma is being incorporated into the response assessment in lymphoma as published by the Imaging Subcommittee of International Harmonization Project in Lymphoma. The exact role of FDG PET in non-Hodgkin's lymphoma (NHL) associated with autologous stem cell transplant (ASCT) is unclear. Numerous studies have identified pretransplant PET scans as being highly prognostic with regard to overall and PFS after ASCT. Many included a wide range of histologies, including Hodgkin's lymphoma and NHL. In studies with mixed histologies, PFS at 2 years has been improved by as much as 82% in patients with negative pre-ASCT PET scans. In studies incorporating only patients with NHL, improvements in failure-free survival have been reported as high as 43% for patients with negative pre-ASCT PET imaging. Limitations have included inclusion of many histologies, different reported time points, small retrospective studies and variation in the interpretation of a positive PET. Validation is ongoing in larger prospective trials. Future directions include the potential incorporation of post-ASCT therapy, such as radiation therapy or maintenance antibody therapy, for patients with positive pre-ASCT PET scans. PMID- 18391992 TI - Rituximab-responsive Guillain-Barre syndrome following allogeneic hematopoietic SCT. PMID- 18391993 TI - Operating room redesign: building safety through a culture of teamwork. PMID- 18391995 TI - Doing better at doing good. AB - Global health initiatives should place greater emphasis on mechanisms for encouraging small and medium-sized biotech enterprises to participate in the fight against neglected diseases. PMID- 18391994 TI - Diabetic eNOS knockout mice develop distinct macro- and microvascular complications. AB - Functional consequences of impaired endothelial nitric oxide synthase (eNOS) activity causing organ-specific abnormalities on a diabetic setting are not completely understood. In this study, we extensively characterized a diabetic mouse model (lepr(db/db)) in which eNOS expression is genetically disrupted (eNOS /-). The eNOS-/-/ lepr(db/db) double-knockout (DKO) mice developed obesity, hyperglycemia, hyperinsulinemia and hypertension. Analysis of tissues from DKO mice showed large islets in the pancreas and fat droplets in hepatocytes. Interestingly, the aorta was normal and atherogenic lesions were not observed. Abnormalities in the aorta including poor re-endothelialization and increased medial wall thickness were evident only in response to deliberate injury. In contrast, significant glomerular capillary damage in the kidney was identified, with DKO mice demonstrating a robust diabetic nephropathy similar to human disease. The vascular and renal impairments in DKO mice were pronounced despite lower fasting plasma glucose levels compared to lepr(db/db) mice, indicating that eNOS is a critical determinant of hyperglycemia-induced organ-specific complications and their severity in diabetes. Results provide the first evidence that absence of eNOS in diabetes has a greater deleterious effect on the renal microvasculature than on the larger aortic vessel. The DKO model may suggest novel therapeutic strategies to prevent both vascular and renal complications of diabetes. PMID- 18391996 TI - A voice with no words. PMID- 18391997 TI - US credit crunch impacts biotech across the globe. PMID- 18391998 TI - Dynogen's SPAC. PMID- 18391999 TI - Can 'double blockbuster' strengthen Amgen's backbone? PMID- 18392000 TI - India's IP snub. PMID- 18392001 TI - Four rebuffs for Cabilly. PMID- 18392002 TI - Companies line up for hereditary angioedema market. PMID- 18392003 TI - Thumbs up for Avastin. PMID- 18392004 TI - Tear-free onions. PMID- 18392005 TI - Drug makers chase cancer stem cells. PMID- 18392006 TI - Amgen's Japan deal. PMID- 18392007 TI - Bush tails follow-ons. PMID- 18392008 TI - Profile: Dan Ravicher. AB - A patent attorney relies on guts, smarts and 'deep throats' to take on what he considers a broken patent system. PMID- 18392009 TI - A mid-life crisis for aging theory. PMID- 18392011 TI - An inconvenient version of events. PMID- 18392012 TI - Transendothelial movement and caveolae. PMID- 18392014 TI - Expanding YIC status. PMID- 18392015 TI - Bt corn in Spain--the performance of the EU's first GM crop. PMID- 18392016 TI - Newtons of the leaves of grass. AB - Certain ethical implications of synthetic biology research go beyond those of genetic engineering. PMID- 18392018 TI - Patenting pluripotence: the next battle for stem cell intellectual property. AB - What will be the impact of induced pluripotent stem cell discoveries on the current stem cell patent landscape? PMID- 18392020 TI - Toward a renewable source of pancreatic beta-cells. PMID- 18392021 TI - Targeting siRNA to arrest fibrosis. PMID- 18392022 TI - Cancer biomarker profiling with microRNAs. PMID- 18392023 TI - The oldest antibodies newly discovered. PMID- 18392024 TI - siRNAs with guts. PMID- 18392026 TI - Discovering microRNAs from deep sequencing data using miRDeep. AB - The capacity of highly parallel sequencing technologies to detect small RNAs at unprecedented depth suggests their value in systematically identifying microRNAs (miRNAs). However, the identification of miRNAs from the large pool of sequenced transcripts from a single deep sequencing run remains a major challenge. Here, we present an algorithm, miRDeep, which uses a probabilistic model of miRNA biogenesis to score compatibility of the position and frequency of sequenced RNA with the secondary structure of the miRNA precursor. We demonstrate its accuracy and robustness using published Caenorhabditis elegans data and data we generated by deep sequencing human and dog RNAs. miRDeep reports altogether approximately 230 previously unannotated miRNAs, of which four novel C. elegans miRNAs are validated by northern blot analysis. PMID- 18392027 TI - Making it stick: convection, reaction and diffusion in surface-based biosensors. AB - The past decade has seen researchers develop and apply novel technologies for biomolecular detection, at times approaching hard limits imposed by physics and chemistry. In nearly all sensors, the transport of target molecules to the sensor can play as critical a role as the chemical reaction itself in governing binding kinetics, and ultimately performance. Yet rarely does an analysis of the interplay between diffusion, convection and reaction motivate experimental design or interpretation. Here we develop a physically intuitive and practical understanding of analyte transport for researchers who develop and employ biosensors based on surface capture. We explore the qualitatively distinct behaviors that result, develop rules of thumb to quickly determine how a given system will behave, and derive order-of-magnitude estimates for fundamental quantities of interest, such as fluxes, collection rates and equilibration times. We pay particular attention to collection limits for micro- and nanoscale sensors, and highlight unexplained discrepancies between reported values and theoretical limits. PMID- 18392030 TI - Almost in bloom. AB - St. Louis wants to become a hub of agricultural biotechnology. All it needs is more start-ups and funds. PMID- 18392032 TI - Genomic fluidity and pathogenic bacteria: applications in diagnostics, epidemiology and intervention. AB - The increasing availability of DNA-sequence information for multiple pathogenic and non-pathogenic variants of individual bacterial species has indicated that both DNA acquisition and genome reduction have important roles in genome evolution. Such genomic fluidity, which is found in human pathogens such as Escherichia coli, Helicobacter pylori and Mycobacterium tuberculosis, has important consequences for the clinical management of the diseases that are caused by these pathogens and for the development of diagnostics and new molecular epidemiological methods. PMID- 18392034 TI - Diatoms in a future ocean--stirring it up. PMID- 18392036 TI - Assessment of the physical activity level with two questions: validation with doubly labeled water. AB - OBJECTIVE: To validate a two-question questionnaire on physical activity with the doubly labeled water (DLW) method. DESIGN: Cross-sectional study. SUBJECTS: Nine volunteers, age 33-75 years, with a mean body mass index (BMI) (kg m(-2)) of 27.4. MEASUREMENTS: A questionnaire with one question on physical activity at work and one question on physical activity during leisure time. The answers were converted into a PAL (physical activity level=energy expenditure/basal metabolic rate) value, which was validated with the DLW method. RESULTS: The mean values (s.d.) of PAL for the questionnaire and DLW measurements were 1.7 (0.1) and 1.7 (0.1), respectively, with a mean difference of 0.004 (0.172). CONCLUSIONS: The results were promising to the extent that they could be used in large-scale epidemiological studies. PMID- 18392037 TI - Human serum levels of fetal antigen 1 (FA1/Dlk1) increase with obesity, are negatively associated with insulin sensitivity and modulate inflammation in vitro. AB - OBJECTIVE: To investigate fetal antigen 1 (FA1) protein within the context of human obesity and its relation with insulin sensitivity. SUBJECTS: Cross sectional study that analyses circulating levels of FA1 in two selected human cohorts: n=127 men for the study of FA1 circulating levels in the context of obesity and insulin sensitivity (S(i)); and n=61 severely obese women before and after bariatric surgery. The response in vitro to FA1 protein on human cell lines of monocytes, preadipocytes and mature adipocytes was studied. MEASUREMENTS: Anthropometrical parameters: body mass index, waist-to-hip ratio, waist circumference, fat-free mass and fat mass. Clinical parameters: lipid profile (high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, total cholesterol, triglycerides), glycemic profile (fasting glucose, insulin, S(i), HOMA-IR (Homeostasis Model Assessment of Insulin Resistance), cytokines (sIL-6), adipokines (adiponectin) and circulating soluble fractions of tumor necrosis factor-alpha receptors 1 and 2 (sTNFR1 and sTNFR2). RESULTS: IN the obesity study, levels of FA1 in serum were found to increase with obesity. The S(i) index was negatively dependent on FA1 levels. In severe obesity, serum levels of FA1 decreased 1.4-fold 6 months after bariatric surgery. In vitro assays with FA1 protein on human monocytes and adipocytes cell lines modified the expression of pro-inflammatory cytokines and adipokines (tumor necrosis factor-alpha (TNFalpha), monocyte chemoattractant protein-1 (MCP-1), IL 6 (interleukin-6) and adiponectin). CONCLUSION: FA1 serum levels were increased in obese subjects and might influence S(i). The stimulatory effect of FA1 protein on pro-inflammatory cytokines on both immune and adipose cell types could contribute to worsening the inflammatory environment observed in obesity. PMID- 18392038 TI - Assessing the capacity of the sympathetic nervous system to respond to a cardiovascular challenge in human spinal cord injury. AB - STUDY DESIGN: Measurement of haemodynamic responses and cutaneous blood flow during an inspiratory-capacity apnoea following spinal cord injury (SCI). OBJECTIVE: To assess the capacity of the sympathetic nervous system to respond to a cardiovascular challenge following SCI. SETTING: Prince of Wales Medical Research Institute, Australia. SUBJECTS: Thirteen spinal cord injured subjects with injuries ranging from C5-T8 and eight able-bodied control subjects. METHODS: Continuous blood pressure, an electrocardiogram, respiration and cutaneous blood flow were recorded during a static maximum inspiratory breath-hold for 40 s. RESULTS: On average, systolic blood pressure decreased 26% from baseline in the spinal group during the breath-hold and remained below baseline throughout the entire apnoeic period. Heart rate in this group had an initial decrease from baseline but quickly increased throughout the breath-hold, being 17% above baseline in the recovery period. Systolic pressure in the control group decreased 12% from baseline at the beginning of the breath-hold but quickly stabilized for the remainder of the apnoea, with heart rate initially decreasing 22% and remaining below baseline throughout the breath-hold. CONCLUSION: A maximal inspiratory breath-hold, which is known to cause a sustained increase in muscle sympathetic nerve activity, is a simple test to perform in supine spinal cord injured subjects, and provides information on the capacity of muscle and splanchnic vasoconstrictor activity to increase blood pressure in SCI. A sustained decrease in blood pressure, coupled with an increase in heart rate, infers interruption of sympathetic vasoconstrictor pathways to muscle and splanchnic vascular beds. PMID- 18392039 TI - Evaluation of cranberry tablets for the prevention of urinary tract infections in spinal cord injured patients with neurogenic bladder. AB - STUDY DESIGN: Randomized, double blind, placebo-controlled trial with a crossover design. OBJECTIVE: To evaluate cranberry tablets for the prevention of urinary tract infection (UTI) in spinal cord injured (SCI) patients. SETTING: Spinal Cord Injury Unit of a Veterans Administration Hospital, MA, USA. METHODS: Subjects with spinal cord injury and documentation of neurogenic bladder were randomized to receive 6 months of cranberry extract tablet or placebo, followed by the alternate preparation for an additional 6 months. The primary outcome was the incidence of UTI. RESULTS: Forty-seven subjects completed the trial. We found a reduction in the likelihood of UTI and symptoms for any month while receiving the cranberry tablet (P<0.05 for all). During the cranberry period, 6 subjects had 7 UTI, compared with 16 subjects and 21 UTI in the placebo period (P<0.05 for both number of subjects and incidence). The frequency of UTI was reduced to 0.3 UTI per year vs 1.0 UTI per year while receiving placebo. Subjects with a glomerular filtration rate (GFR) greater than 75 ml min(-1) received the most benefit. CONCLUSION: Cranberry extract tablets should be considered for the prevention of UTI in SCI patients with neurogenic bladder. Patients with a high GFR may receive the most benefit. SPONSORSHIP: Spinal Cord Research Foundation, sponsored by the Paralyzed Veterans of America. PMID- 18392040 TI - Synergistic antitumor effects of CpG oligodeoxynucleotide and STAT3 inhibitory agent JSI-124 in a mouse melanoma tumor model. AB - One of the major limitations for cancer immunotherapy is related to the frequent existence of an intra-tumoral immunosuppressive environment, to which STAT3 (Signal transducer and activator of transcription-3) activation in tumor and dendritic cells (DCs) are believed to contribute. In this study, we tested the hypothesis that the combination of CpG (a DC activator) and JSI-124 (a STAT3 inhibitor) may generate synergistic antitumor effects compared to CpG or JSI-124 alone. B16-F10, a mouse melanoma cell line that has constitutively active STAT3, was grafted in C57BL/6 mice and then tumor-bearing mice treated intra-tumorally with (a) phosphate buffered saline, (b) 10 microg CpG, (c) 1 mg kg(-1) JSI-124 or (d) 10 microg CpG+1 mg kg(-1) JSI-124. The effects of treatments on tumor growth, survival and antitumor immune responses were evaluated. Although significant antitumor effects were detected with the single-agent treatments, the CpG+JSI-124 treatment resulted in synergistic antitumor effects compared to CpG or JSI-124 alone. Correlating with these findings, the combination therapy resulted in significantly higher intra-tumoral levels of several proinflammatory, TH1-related cytokines (including IL-12, IFN-gamma, TNF-alpha and IL-2), increases in intra tumoral CD8+ and CD4+ T cells expressing activation/memory markers and NK cells and increases in activated DCs in the tumors and regional lymph nodes (LNs). Concomitantly, the combination therapy led to a significantly decreased level of immunosuppression, as evidenced by lower intra-tumoral level of VEGF and TGF beta, and decreased number of CD4+CD25+Foxp3+ regulatory T cells in the regional LNs. This study has provided the proof-of-principle for combining CpG and JSI-124 to enhance antitumor immune responses. PMID- 18392041 TI - Determination of T-cell fate by dendritic cells: a new role for asymmetric cell division? AB - The production, from a single naive T cell, of the many different activated T cell types required for an effective immune response has fascinated immunologists for decades. This process underpins the development of vaccines, immunosuppressive regimes in transplant patients, and immunotherapy in cancer among other things. Despite the enormous advances in detailing the mechanisms and influencing factors in the differentiation of each T-cell subtype, it is still not clear how the different T-cell progeny are produced in proportions that are appropriate for each situation. This review discusses the notion that asymmetric cell division might allow for the regulated generation of different cell populations. PMID- 18392043 TI - Regulating inflammation: the ying and yang of NF-kappaB activation. PMID- 18392042 TI - Geography and plumbing control the T cell response to infection. AB - The orchestrated movement of cells of the immune system is essential to generation of productive responses leading to protective memory development. Recent advances have allowed the direct microscopic visualization of lymphocyte and antigen-presenting cell migration and interaction during immune response initiation and progression. These studies have defined important characteristics of the microanatomy of lymphocyte movement, particularly in the lymph node. Moreover, the ability to track endogenous antigen-specific T cells has revealed a coordinated pathway of CD8 T cell movement in the spleen following primary and secondary infection. As a consequence, the local anatomy of secondary lymphoid tissues during infection has emerged as a critical regulator of immunity. While some of the factors responsible for the migratory cues instructing immune cell movement have been identified, much remains to be learned. Here, we provide a brief overview of studies examining CD8 T cell localization during the immune response to infection in the context of our current understanding of immune system structure. PMID- 18392044 TI - Blood monocytes: distinct subsets, how they relate to dendritic cells, and their possible roles in the regulation of T-cell responses. AB - Monocytes can have important effects on the polarization and expansion of lymphocytes and may contribute to shaping primary and memory T-cell responses in humans and mice. However, their precise contribution in terms of cellular subsets and the molecular mechanisms involved remains to be determined. Mouse monocytes originate from a bone marrow progenitor, the macrophage and DC precursor (MDP), which also gives rise to conventional dendritic cells through a separate differentiation pathway. Mouse monocytes may be grouped in different functional subsets. The CD115(+) Gr1(+) 'inflammatory' monocyte subset can give rise not only to immunostimulatory 'TipDCs' in infected mice but also to immunosuppressive 'myeloid-derived suppressor cells' in tumor-bearing mice. CD115(+) Gr1(+) monocytes can also contribute to the renewal of several resident subsets of macrophages and DCs, such as microglia and Langerhans cells, in inflammatory conditions. The CD115(+) Gr1(-) 'resident' monocyte subset patrols blood vessels in the steady state and extravasates during infection with Listeria monocytogenes or in the healing myocardium. CD115(+) Gr1(-) monocytes are responsible for an early and transient inflammatory burst during Lm infection, which may play a role in the recruitment of other effector cells and subsequently differentiate toward 'M2'-like macrophages that may be involved in wound healing. More research will no doubt confirm the existence of more functional subsets, the developmental relationship between mouse subsets as well as the correspondence between mouse subsets and human subsets of monocytes. We will discuss here the potential roles of monocytes in the immune response, the existence of functional subsets and their relationship with other myeloid cells, including dendritic cells. PMID- 18392045 TI - TM1 and TM2: two mutant alleles that are involved in the pre-TCR/TCR signaling. AB - It has been shown that the transgene insertional mutations TM1 and TM2 constitute a genetic trait controlling thymocyte development. Here we conducted a detailed analysis of the impact of TM1 and TM2 double mutation on thymocyte development. We found that the hemizygous TM1 and TM2 double transgenic mice possessed much smaller thymi. Flow cytometric analysis revealed a severe blockage of T-cell development at the transition from DN3 to DN4 stage and pre-T-cell receptor (pre TCR)/TCR signaling appeared to be impaired. We could not identify any known gene that was implicated in a similar function in the chromosomal regions 7E-F1 and 11B5-C, where TM1 and TM2 mutations were mapped to respectively. Thus, TM1 and TM2 mutations represent two novel alleles that define a genetic trait controlling DN3 thymocyte development, possibly through modulating the signals downstream of the pre-TCR. PMID- 18392046 TI - Distress and social dysfunction following prostate cancer treatment: a longitudinal cross-cultural comparison of Japanese and American men. AB - We assessed the impact of bother with urinary and bowel dysfunction on social activities among men in Japan and the United States following primary therapy for localized prostate cancer. In paired longitudinal outcomes studies, we measured general and disease-specific health-related quality of life in 400 Japanese and 427 American men who underwent radical prostatectomy or brachytherapy for localized prostate cancer. Outcomes included the social function domain of the Medical Outcomes Study Short Form-36 and the University of California, Los Angeles Prostate Cancer Index, all of which are scored 0-100. Participants completed the questionnaires before and 1, 12 and 24 months after treatment. Among men who reported any urinary bother, Japanese men had slightly better urinary function than American men (84 vs 77, P<0.01). Before brachytherapy, urinary bother was weakly correlated with social function in both the countries; after brachytherapy, urinary bother was strongly correlated with social function in American but not Japanese men. After brachytherapy, bowel dysfunction had a stronger correlation with social function in American than Japanese men (P<0.05). The bother associated with urinary and bowel dysfunction after surgery or brachytherapy for prostate cancer has a greater impact on social function in American men than in Japanese men. PMID- 18392047 TI - Impaired functions of neural stem cells by abnormal nitric oxide-mediated signaling in an in vitro model of Niemann-Pick type C disease. AB - Nitric oxide (NO) has been implicated in the promotion of neurodegeneration. However, little is known about the relationship between NO and the self-renewal or differentiation capacity of neural stem cells (NSCs) in neurodegenerative disease. In this study, we investigated the effect of NO on self-renewal of NSCs in an animal model for Niemann-Pick type C (NPC) disease. We found that NO production was significantly increased in NSCs from NPC1-deficient mice (NPC1-/ ), which showed reduced NSC self-renewal. The number of nestin-positive cells and the size of neurospheres were both significantly decreased. The expression of NO synthase (NOS) was increased in neurospheres derived from the brain of NPC1-/- mice in comparison to wild-type neurospheres. NO-mediated activation of glycogen synthase kinase-3beta (GSK3beta) and caspase-3 was also observed in NSCs from NPC1-/- mice. The self-renewal ability of NSCs from NPC1-/- mice was restored by an NOS inhibitor, L-NAME, which resulted in the inhibition of GSK3beta and caspase-3. In addition, the differentiation ability of NSCs was partially restored and the number of Fluoro-Jade C-positive degenerating neurons was reduced. These data suggest that overproduction of NO in NPC disease impaired the self-renewal of NSCs. Control of NO production may be key for the treatment of NPC disease. PMID- 18392048 TI - Wnt signaling and stem cell control. AB - Wnt signaling has been implicated in the control over various types of stem cells and may act as a niche factor to maintain stem cells in a self-renewing state. As currently understood, Wnt proteins bind to receptors of the Frizzled and LRP families on the cell surface. Through several cytoplasmic relay components, the signal is transduced to ss-catenin, which then enters the nucleus and forms a complex with TCF to activate transcription of Wnt target genes. Wnts can also signal through tyrosine kinase receptors, in particular the ROR and RYK receptors, leading to alternative modes of Wnt signaling. During the growth of tissues, these ligands and receptors are dynamically expressed, often transcriptionally controlled by Wnt signals themselves, to ensure the right balance between proliferation and differentiation. Isolated Wnt proteins are active on a variety of stem cells, including neural, mammary and embryonic stem cells. In general, Wnt proteins act to maintain the undifferentiated state of stem cells, while other growth factors instruct the cells to proliferate. These other factors include FGF and EGF, signaling through tyrosine kinase pathways. PMID- 18392049 TI - Perinatal risk factors for hepatoblastoma. AB - We confirmed the strong association of hepatoblastoma with very low birth weight (relative risk <1000 g vs >or=2000 g=25.6; 95% confidence interval: 7.70-85.0) and demonstrated independent associations with congenital abnormalities and maternal Asian race in a population-based Minnesota study that included 36 cases and 7788 controls. PMID- 18392051 TI - A gastrin transcript expressed in gastrointestinal cancer cells contains an internal ribosome entry site. AB - As the hormone gastrin promotes gastrointestinal (GI) cancer progression by triggering survival pathways, regulation of gastrin expression at the translational level was explored. Sequence within the 5' untranslated region of a gastrin transcript expressed in GI cancer cells was investigated, then cloned into a bicistronic vector upstream of firefly luciferase and transfected into a series of GI cancer cell lines. Firefly luciferase activity was measured relative to that of a cap-dependent Renilla luciferase. A gastrin transcript that was different from that described in Ensembl was expressed in GI cancer cells. Its transcription appears to be initiated within the region designated as the gene's first intron. In GI cancer cells transfected with the bicistronic construct, firefly luciferase activity increased 8-15-fold compared with the control vector, and there was a further induction of the signal (up to 25-fold) following exposure of the cells to genotoxic stress or hypoxia, suggesting that the sequence acts as an internal ribosome entry site. These data suggest that the gastrin transcript within GI cancer cells contains an internal ribosome entry site that may allow continued expression of gastrin peptides when normal translational mechanisms are inactive, such as in hypoxia, thereby promoting cancer cell survival. PMID- 18392050 TI - Early diagnosis of pancreatic cancer: neutrophil gelatinase-associated lipocalin as a marker of pancreatic intraepithelial neoplasia. AB - Pancreatic cancer is a highly lethal malignancy with a dismal 5-year survival of less than 5%. The scarcity of early biomarkers has considerably hindered our ability to launch preventive measures for this malignancy in a timely manner. Neutrophil gelatinase-associated lipocalin (NGAL), a 24-kDa glycoprotein, was reported to be upregulated nearly 27-fold in pancreatic cancer cells compared to normal ductal cells in a microarray analysis. Given the need for biomarkers in the early diagnosis of pancreatic cancer, we investigated the expression of NGAL in tissues with the objective of examining if NGAL immunostaining could be used to identify foci of pancreatic intraepithelial neoplasia, premalignant lesions preceding invasive cancer. To examine a possible correlation between NGAL expression and the degree of differentiation, we also analysed NGAL levels in pancreatic cancer cell lines with varying grades of differentiation. Although NGAL expression was strongly upregulated in pancreatic cancer, and moderately in pancreatitis, only a weak expression could be detected in the healthy pancreas. The average composite score for adenocarcinoma (4.26+/-2.44) was significantly higher than that for the normal pancreas (1.0) or pancreatitis (1.0) (P<0.0001). Further, although both well- and moderately differentiated pancreatic cancer were positive for NGAL, poorly differentiated adenocarcinoma was uniformly negative. Importantly, NGAL expression was detected as early as the PanIN-1 stage, suggesting that it could be a marker of the earliest premalignant changes in the pancreas. Further, we examined NGAL levels in serum samples. Serum NGAL levels were above the cutoff for healthy individuals in 94% of pancreatic cancer and 62.5% each of acute and chronic pancreatitis samples. However, the difference between NGAL levels in pancreatitis and pancreatic cancer was not significant. A ROC curve analysis revealed that ELISA for NGAL is fairly accurate in distinguishing pancreatic cancer from non-cancer cases (area under curve=0.75). In conclusion, NGAL is highly expressed in early dysplastic lesions in the pancreas, suggesting a possible role as an early diagnostic marker for pancreatic cancer. Further, serum NGAL measurement could be investigated as a possible biomarker in pancreatitis and pancreatic adenocarcinoma. PMID- 18392052 TI - Prevalence of type-specific HPV infection by age and grade of cervical cytology: data from the ARTISTIC trial. AB - Human papillomavirus (HPV) infection causes cervical cancer and premalignant dysplasia. Type-specific HPV prevalence data provide a basis for assessing the impact of HPV vaccination programmes on cervical cytology. We report high-risk HPV (HR-HPV) type-specific prevalence data in relation to cervical cytology for 24,510 women (age range: 20-64; mean age 40.2 years) recruited into the ARTISTIC trial, which is being conducted within the routine NHS Cervical Screening Programme in Greater Manchester. The most common HR-HPV types were HPV16, 18, 31, 51 and 52, which accounted for 60% of all HR-HPV types detected. There was a marked decline in the prevalence of HR-HPV infection with age, but the proportion due to each HPV type did not vary greatly with age. Multiple infections were common below the age of 30 years but less so between age 30 and 64 years. Catch up vaccination of this sexually active cohort would be expected to reduce the number of women with moderate or worse cytology by 45%, but the number with borderline or mild cytology would fall by only 7%, giving an overall reduction of 12% in the number of women with abnormal cytology and 27% in the number with any HR-HPV infection. In the absence of broader cross-protection, the large majority of low-grade and many high-grade abnormalities may still occur in sexually active vaccinated women. PMID- 18392053 TI - Switching to letrozole or exemestane improves hot flushes, mood and quality of life in tamoxifen intolerant women. AB - We report an open-label, prospective, crossover study involving 184 post menopausal women experiencing hot flushes on adjuvant tamoxifen (T). Six weeks after switching to an AI, the primary end point, hot flush score, improved by 47.3% (P<0.001) compared to those reported on T. The mean mood rating scale (MRS) score improved by 9.7% (P=0.01). The total mean combined FACT (b+es) score improved from 134.2 (95% CI +/-2.96) to 143.5 (95% CI +/-2.96 <0.001), and the endocrine subscale improved by 9.8% from 51.73 (95% CI +/-1.38) to 57.34 (CI +/ 1.38, P<0.001). At 6 weeks, significantly more women chose to remain on an AI: 133 (72%), vs 40 (22%) (P<0.001) preferring T. At 3 months, 107 (58%) preferred to remain on an AI, 55(30%) on T, and 22 (12%) withdrew. The overall arthralgia rate at 3 months was 47% on AI and 30% on T (P=0.001). In all 182 (99%) women reported appreciating the opportunity to experience both drugs. These data suggest that if patients suffering significant adverse effects on T are given the opportunity to try an AI, this empowers them to prioritize relative side-effects, improving wellbeing in a significant proportion. These data also highlight the need for hospital follow-up in this intolerant cohort. PMID- 18392056 TI - Children in New Zealand: their health and human rights. PMID- 18392055 TI - Mutational analysis of oncogenic AKT E17K mutation in common solid cancers and acute leukaemias. AB - Mounting evidence indicates that alterations of AKT signalling play important roles in cancer development. An earlier study discovered an oncogenic AKT1 gene mutation (AKT1 E17K) in breast, colorectal and ovarian cancers. The aim of this study was to see whether the AKT1 E17K mutation is common in breast, colorectal, lung, gastric and hepatocellular carcinomas and acute leukaemias. We analysed the presence of the AKT1 E17K mutation in 731 cancer tissues by a single-strand conformation polymorphism assay. In addition, we analysed the corresponding sequences of AKT1 E17K in AKT2 and AKT3 genes. Overall, we detected the four AKT1 E17K mutations in the breast cancers (4/93; 4.3%), but none in other cancers. There was no AKT2 or AKT3 mutation in the cancers. This study demonstrated that the AKT1 E17K mutation occurs in breast cancers at a low frequency, and that it is rare in other common cancers, including colorectal, lung, gastric and hepatocellular carcinomas and acute leukaemias. Despite the confirmed oncogenic function of the AKT1 E17K, the rare incidences of the mutation suggest that it may not play a crucial role in the development of the most common types of human cancers. PMID- 18392054 TI - The role of early life genistein exposures in modifying breast cancer risk. AB - Review of the existing literature suggests that consumption of soy foods or an exposure to a soy isoflavone genistein during childhood and adolescence in women, and before puberty onset in animals, reduces later mammary cancer risk. In animal studies, an exposure that is limited to the fetal period or adult life does not appear to have the same protective effect. A meta-analysis of human studies indicates a modest reduction in pre- and postmenopausal risk when dietary intakes are assessed during adult life. These findings concur with emerging evidence indicating that timing may be vitally important in determining the effects of various dietary exposures on the susceptibility to develop breast cancer. In this review, we address the mechanisms that might mediate the effects of an early life exposure to genistein on the mammary gland. The focus is on changes in gene expression, such as those involving BRCA1 and PTEN. It will be debated whether mammary stem cells are the targets of genistein-induced alterations and also whether the alterations are epigenetic. We propose that the effects on mammary gland morphology and signalling pathways induced by pubertal exposure to genistein mimic those induced by the oestrogenic environment of early first pregnancy. PMID- 18392057 TI - The knock-on effects of unrestrained drinking. PMID- 18392058 TI - Role of alcohol in maxillofacial fractures. AB - BACKGROUND: Excessive consumption of alcohol results in impaired judgement and inappropriate behaviour, and is often a major contributor to interpersonal violence and motor vehicle accidents. This study examines the experience of a tertiary centre in alcohol-related facial fractures. METHODS: A retrospective database of patients presenting to the Oral and Maxillofacial Surgery Service at Christchurch Hospital (New Zealand) during an 11-year period was reviewed. Variables examined include demographics, type of fractures, mode of injury, and treatment delivered. RESULTS: 2581 patients presented with facial fractures during the study period, 49% of these being alcohol-related. Males accounted for 88% of alcohol-related fractures and 59% were males in the 15 to 29 year age group; 78% of alcohol-related fractures were due to interpersonal violence and 13% to motor vehicle accidents; 65% required hospital admission and 58% underwent surgery. CONCLUSION: The majority of alcohol-related facial fractures were due to interpersonal violence, with young men in the 15 to 29 year age group being predominantly affected. Alcohol-related fractures were associated with an increase in the incidence of hospitalisation and surgery. The high prevalence of alcohol as a contributing factor to facial fractures indicates a need to push for community awareness and public education on the harmful effects of alcohol. PMID- 18392059 TI - Under-18 year old callers to New Zealand's Quitline. AB - AIM: To investigate the characteristics of under-18 year old callers to New Zealand's Quitline (smoking-cessation telephone counselling service). METHODS: Analysis of routinely collected demographic and smoking history characteristics of under-18 year old Quitline callers in 2004 and 2005. RESULTS: In the 24 months of 2004-2005, 2371 under-18s called Quitline (for the first time) seeking smoking cessation support. Females (58.9%) and teens in their older teen years called most often. Compared with adult callers, there were significantly higher proportions of Maori (32.9% vs 19.6%) and Pacific (5% vs 3.6%) under-18 callers, and fewer European (64.0% vs 74.6%) and 'Other' (6.0% vs 7.1%) callers. Despite similar levels of nicotine dependence in under-18 and adult callers (70.1% vs 71.4% reported smoking within 30 minutes of waking), under-18s were issued nicotine replacement therapy (NRT) half as often (RR=0.51). Under-18s were more likely than adults to register a mobile phone number (48.9% vs 44.4%). CONCLUSIONS: Under-18 year old smokers are under-represented in the Quitline calling population. Maori and Pacific under-18s require further cessation support to avoid exacerbating existing disparities in smoking. Awareness that under-18 nicotine dependence is equivalent to that of adults should lead to improved provision of NRT for adolescents. Initiatives involving mobile phone technology are particularly appropriate for improving access to information and treatment for under-18s. Adolescent tobacco cessation should be accorded greater priority in tobacco control policy, practice, and research. PMID- 18392060 TI - "If everyone does it, it's not a big deal." Young people talk about chlamydia testing. AB - AIMS: This study aimed to explore young people's attitudes to chlamydia testing. Data were gathered to inform the development of a clinical trial aimed at increasing chlamydia testing among 16-24 year olds. METHODS: Four single sex focus groups were conducted with 16-24 year old males and females (n=28), and one with health professionals working with this age-group (n=7). A semi-structured interview schedule was used to discuss barriers to chlamydia testing, methods of accessing testing, communicating information about chlamydia and ideas about ways to encourage testing. RESULTS: Reasons for not seeking testing included fear, stigma, denial of personal risk, and a lack of knowledge about chlamydia and about testing procedures. Better education and a need to 'normalise' testing were suggested as ways to increase test-uptake. Preferences for places to seek testing varied among participants, but all groups supported routinely offered chlamydia testing when visiting the doctor for other reasons. Participants also favoured the concept of home-testing. CONCLUSIONS: Young people identified a number of barriers to chlamydia testing, as well as ways to increase testing. These findings can be used to inform the development of much needed new initiatives to control chlamydia in New Zealand. PMID- 18392061 TI - Herpes simplex type 1 versus Herpes simplex type 2 in anogenital herpes; a 10 year study from the Waikato region of New Zealand. AB - AIM: To determine the proportion of Herpes simplex type 1 vs Herpes simplex type 2, as a cause of anogenital herpes in the Waikato region. We specifically looked for changes in the proportions over time, and for gender or age group associations. METHOD: We undertook a retrospective data-analysis of all anogenital isolates positive for Herpes simplex types 1 or 2 in those greater than and equal to 14 years of age, received at Waikato Hospital Laboratory (Hamilton, New Zealand) over a 10-year period. Around half way through the study period, the test type changed from viral culture to HSV nucleic acid amplification assay. RESULTS: Between 1997 and 2006, 3933 anogenital isolates were positive for either Herpes simplex types 1 or 2. The proportion of type 1 vs type 2 oscillated around 30 to 40% over the 10 years, with no particular trend. Overall, 71% of positive isolates were from females, with 80% of the type 1 isolates from females. Presence of type 1 was inversely proportional to age, accounting for 53% of positive isolates in the <25 year age group, 30% in the 25 35 year group, and 26% in the >35 year group. CONCLUSION: Herpes simplex type 1 is a major contributor to anogenital herpes in the Waikato. Females accounted for the majority of both types and there was a much higher proportion of type 1 seen in the younger age groups. These findings have been reported overseas but this is the first reported New Zealand data. PMID- 18392062 TI - Walking to school: frequency and predictors among primary school children in Dunedin, New Zealand. AB - AIM: To estimate the frequency of walking to school among primary school children and examine associated factors. METHOD: In-class survey of Years 1-6 children attending Dunedin primary schools, November 2004, and a take home, written questionnaire for parents and caregivers. RESULTS: On study day, 34.5% of children walked to school and 36.8% intended to walk home. Overall, 1157 completed caregiver questionnaires were returned (68%) indicating that 47.5% of children walked to or from school less than three times per week. The strongest predictor of walking was proximity to school (<1 km OR 29.3, 1-2 km OR 7.7, 2-3 km OR 3.0, >3 km OR 1.00). Other predictors were not having a car in the household (OR 10.9), attending a (low socioeconomic) decile 2 to 4 school (2.35), having three or more adults in the household (1.85), being in a higher school year (1.72), having non-New Zealand European ethnicity (>1.41), having a parent who had walked to school (1.35), and being male (1.33). CONCLUSIONS: This study established a baseline for the percentage of Dunedin primary school children walking to school. Key potentially modifiable predictors of walking were proximity to school and not having a car in the household. These findings have implications for health, transport and educational policies. PMID- 18392063 TI - Rheumatic fever diagnosis, management, and secondary prevention: a New Zealand guideline. AB - AIM: The National Heart Foundation of New Zealand, and the Cardiac Society of Australia and New Zealand (CSANZ) recently launched an evidence-based review and guideline entitled New Zealand Guideline for Rheumatic Fever Diagnosis, Management, and Secondary Prevention. This paper is a brief summary. METHOD: This Guideline was developed by a writing group comprising experts in the area. Relevant literature was identified and reviewed, and the Australian guideline for rheumatic fever and rheumatic heart disease was reviewed and adapted for the New Zealand context. A peer review and stakeholder consultation process followed the development of the draft document. RESULTS: The final draft of the New Zealand guideline was endorsed by Te Hotu Manawa Maori, Pacific Islands Heart Beat, The Paediatric Society of New Zealand, and the Rheumatic Fever Trust of New Zealand- plus approved by a number of organisations including the Royal Australasian College of Physicians, the Australasian Society for Infectious Disease, the Pasifika Medical Association, and Te Ohu Rata o Aotearoa. Two subsequent New Zealand guidelines for rheumatic fever: Sore Throat Management and Primary Prevention are also in production. The complete guideline, and associated summary algorithms, can be downloaded from www.nhf.org.nz CONCLUSION: A New Zealand Guideline for Rheumatic Fever Diagnosis, Management and Secondary Prevention of Acute Rheumatic Fever should result in improved consistency in the approach to this disease, and reduced mortality and morbidity from acute rheumatic fever and rheumatic heart disease. PMID- 18392064 TI - Rhabdomyolysis in a glue sniffer. PMID- 18392065 TI - Spontaneous passage of large ureteric stone in a child. PMID- 18392066 TI - The changing epidemiology of food allergy--implications for New Zealand. AB - Food allergy (FA) is recognised as an important public health problem in developed countries. Recent studies suggest a significant proportion of the general population has a definable FA. The methods used to study FA influence published estimates of incidence and prevalence. In particular, studies relying on self-assessment are likely to overestimate the condition compared to studies using a comprehensive approach including symptoms, allergy testing, rigorously conducted laboratory tests, and food challenges. Currently there are no reliable data on the prevalence of FA in New Zealand. This has had several adverse consequences including the lack of public hospital services for patients with severe allergies. In this article we summarise the epidemiological data on FA and discuss the implications for New Zealand. PMID- 18392067 TI - Mental health services for children and adolescents in New Zealand, outcomes, and the Health of the Nation Outcome Scale for Children and Adolescents (HoNOSCA). AB - Patients and their families as well as communities, service providers, and funders of services would be united in their desire that children and adolescents who require mental health services should receive those services. There would also be agreement that treatment delivered by these services should be safe, effective, and (given that resources for these mental health services are limited) delivered in a timely and cost-effective manner. Furthermore, there would be a consensus that outcomes of treatment are extremely important and that there is a need to evaluate these in a valid manner. This article reviews current access to Child and Adolescent Mental Health Services (CAMHS) in New Zealand as well as issues relevant to the introduction of routine outcome measurement in these services; and critically appraises the psychometric properties and clinical utility of the first routine outcome measure introduced for CAMHS by the Ministry of Health (MOH)--the Health of the Nation Outcome Scale for Children and Adolescents (the HoNOSCA). It is argued that the evidence base for the implementation of routine outcome measurement is poor, that systematic evaluation of its introduction should occur, and that already under-funded CAMHS should be adequately resourced to support the additional work involved. PMID- 18392068 TI - Medical image. Hydrocolpos in a child. PMID- 18392069 TI - Medical image. A prepubescent mass. Swyer syndrome with a bilateral dysgerminoma. PMID- 18392070 TI - Increased advertising of medicines on New Zealand television since 2001. PMID- 18392071 TI - SSRI antidepressants for adolescents: Professor Werry replies to Medsafe. PMID- 18392073 TI - Do doctors in the First World have an obligation to help their counterparts in the developing World? PMID- 18392072 TI - Bacteraemia at Gisborne hospital. PMID- 18392081 TI - Immunocytochemical study on microtubule reorganization in HL-60 leukemia cells undergoing apoptosis. AB - BACKGROUND: Microtubules (MT) are important components of cell cytoskeleton and play key roles in cell motility mitosis and meiosis. They are also the targets of several anticancer agents which indicating their importance in maintaining cell viability. Microtubular reorganization contributing to apoptotic morphology occurs in normal and neoplastic cells undergoing apoptosis induced by cytotoxic drugs. The aim of this study was to correlate the changes in the MT with behavior of the gamma-tubulin in apoptotic cell, and to see if apoptitic MT showed biochemical characteristics of stable MT. METHODS: Apoptosis was induced in the human leukemia cells (HL-60) by treatment with 1 microM of all-trans retinoic acid over a 5-day period. The time course of changes was assessed using flow cytometry, DNA fragmentation and immunocytochemistry in cells labeled for alpha tubulins, acetylated alpha-tubulin and gamma-tubulin. RESULTS: The results indicated that gamma-tubulin content is increased after cells have gone through the apoptosis with a diffuse cytoplasmic pattern. Alpha-tubulin did not reveal any specific pattern of polymerization in apoptotic cells and acetylated alpha tubulin content was also decreased in comparison with non-apoptotic cells. CONCLUSION: Our results support the idea that microtubule reorganization is an important factor of the mammalian cells response to apoptosis, and the altered properties of the MT did not reflect changes in function as apoptosis progresses. PMID- 18392082 TI - G1 phase arrest and apoptosis induction in human thyroid cancer cell line Thr.C1.PI33 by 3-hydrogenkwadaphnin isolated from Dendrostellera lessertii. AB - BACKGROUND: Dendrostellera lessertii (Thymelaeaceae) is a toxic plant that grows in parts of Iran. The anti-proliferative properties of its crude methanol extract and one of its active components, 3-hydrogenkwadaphnin (3-HK), have been established using several cancer cell lines. METHODS: In a further attempt to determine the mode of action, two groups of synchronously growing cells were treated with a single dose of 3-HK (3.5 nM) and/or a single dose of the crude extract (equivalent to 0.36 mg plant powder). Every 8 hours, the percentages of cells within G1, S, and G2-M phases were determined by flow cytometric (FCM) analysis; electron microscopic pictures were taken after fixation with 2% glutaraldehyde. RESULTS: Twelve hours after treatments, apoptotic cell death was confirmed by the observation of marked morphological changes of the plasma membrane as microvillar disappearance and the appearance of apoptotic bodies in the treated cells. FCM analyses revealed that the G1 phase arrest was under the influence of the pure substance. CONCLUSION: The results confirmed the previously drawn conclusion that the raw material and the pure substance from D. lessertii exert their anti-tumor effects through cell cycle arrest at G1 phase and diversion of cell fate toward programmed cell death. PMID- 18392083 TI - Suppression of telomerase activity by pyrimethamine: implication to cancer. AB - BACKGROUND: Although pyrimethamine (Tindurin) appears to be effective in the prevention and treatment of some infectious diseases, very little information exists on its unpredictable properties. We design this study to evaluate its anti tumoral effect on a model of cell line. METHODS: The cytotoxic influence of Pyrimethamine on prostate cell line was investigated using an in vitro colometric assay. The potential modulatory effects on metastasis, apoptosis, and immortality characteristics of cells were assessed with gelatin zymography, terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay and telomeric repeat amplification protocol, respectively. RESULTS: Cytotoxicity analysis of pyrimethamine revealed a dose-dependent fashion. An apoptotic influence of pyrimethamine was also confirmed by data obtained from TUNEL assay. Dose-dependent inhibitory effect on matrix metalloproteinases (MMP) was seen in pyrimethamine. A potent inhibitory effect of pyrimethamine was also established by data achieved from TRAPeze telomerase detection kit. CONCLUSIONS: Collectively, as induction of apoptosis together with MMP and telomerase inhibition could be indicative of cancer treatment, pyrimethamine might be considered as a chemopreventative agent in cancer. PMID- 18392084 TI - Buthionine sulfoximine inhibits cytopathic effects and apoptosis induced by infection with AIK-HDC strain of measles virus. AB - BACKGROUND: Measles virus (MV) is a highly contagious agent which causes a major health problem in developing countries. We studied the effect of buthionine sulfoximine (BSO) on the replication of an AIK-HDC strain of MV and its induced apoptosis in Vero cell lines. METHODS: In this study, toxicity of BSO on Vero cells was investigated first, resulted in determination of sub-lethal or non toxic concentration zone of BSO for cells. Next, anti-viral effect of BSO at various time limits was evaluated and virus titer was determined at each stage either as 50% tissue culture infective dose (TCID) 50 or by plaque assay method. Using specific anti-measles IgG, anti-viral effect of BSO on MV replication cycle was evaluated through indirect immunofluorescence assay, meanwhile presence of viral RNA was investigated by RT-PCR and gel electrophoresis. RESULTS: According to the experiments, BSO, at concentration of 50 microM, markedly inhibited the cytopathic effect (CPE) induced by MV. BSO also significantly inhibited apoptosis induced by MV. BSO either influences replication of MV genome, or may inhibit virion formation. CONCLUSION: These results suggest that the inhibition of CPE and apoptosis by BSO induced by MV may be associated with the effect of BSO on viral RNA genome. Therefore, it is suggested that MV infections can induce apoptosis through the activation of a common pathway that can be inhibited by BSO. PMID- 18392085 TI - The beneficial effect of (-)-epigallocatechin-3-gallate in an experimental model of Alzheimer's disease in rat: a behavioral analysis. AB - BACKGROUND: Progressive cognitive decline is one of the hallmark symptoms of Alzheimer's disease (AD) which can be modeled by beta-amyloid injection into specific regions of brain. Since epigallocatechin-3-gallate (EGCG) is a potent antioxidant agent which its role against oxidative stress and inflammation has been shown in prior studies, we tried to determine whether EGCG administration protects against beta-amyloid-induced memory and coordination impairment in rats. METHODS: Animals (male Wistar rats) were divided into four groups: sham operated, EGCG-pretreated sham operated (sham+EGCG), untreated lesion (lesion), and EGCG pretreated lesion (lesion+EGCG). Animals in lesion, lesion+EGCG, and sham+EGCG groups received sterile saline or saline plus EGCG (10 mg/kg) intraperitoneally one day pre-surgery and every other day for three weeks. The lesion was induced one day after EGCG pretreatment by injection of 4 microl of sterile saline or water containing 2 nmol/microl beta-amyloid (1-40) into the hippocampal fissure. For behavioral analysis, psychomotor coordination (PMC) index and spontaneous alternation behavior were assessed using Rota-rod Treadmill and Y-maze, respectively at the third week post-lesion. RESULTS: We found that beta-amyloid (1-40) injection into hippocampus can decrease these behavioral indexes in lesion group in comparison with sham group which is similar to behavioral changes in AD. On the other hand, pretreatment with EGCG can improve the PMC index and spatial Y maze alternation in the lesion+EGCG group in comparison with lesion group. CONCLUSION: We concluded that EGCG can be effective in restoring beta-amyloid induced behavioral derangements in rats regarding coordination and memory abilities. PMID- 18392086 TI - Comparison of embryo development between intracytoplasmic and Piezo-assisted sperm injection after treating mouse sperms by Ca2+ ionophore. AB - BACKGROUND: The purpose of this study was to evaluate the efficiency of intracytoplasmic sperm injection (ICSI) and Piezo-assisted sperm injection after pretreatment with calcium ionophore (CaI) on the mouse embryo development. METHODS: In this study, the conventional ICSI and Piezo-ICSI procedures were used. The efficacy of the methods was examined after mouse matured oocytes were fertilized with or without CaI-treated sperms. RESULTS: Piezo-ICSI demonstrated significantly more favorable results, with a fertilization rate of 64% (conventional ICSI: 42%, P<0.001) and a cleavage rate of 73% (conventional ICSI: 58%, P<0.05). When the Piezo-ICSI procedure was performed with CaI-pretreated sperms, the cleavage rate significantly increased (92% vs. 73%, P<0.05). However, the fertilization rate did not change significantly (64% vs. 56%). CONCLUSION: The Piezo-ICSI accompanies with CaI-treated sperms is more efficient than the conventional ICSI method for fertilizing and thus obtaining more mouse embryos. PMID- 18392087 TI - Biochemistry of pyrethroid resistance in German cockroach (Dictyoptera, Blatellidae) from hospitals of Sari, Iran. AB - BACKGROUND: The German cockroach is an important household insect pest mechanically involved in transmission of a variety of diseases to humans. Different classes of insecticides have extensively been used for its control leading to insecticide resistance development. Hence, for an optimal control of this pest, the status and underlying mechanisms of insecticide resistance should be studied in this group of insects. METHODS: Adult German cockroaches were collected from Imam and Bouali Cina Hospitals (Sari, Iran) and subjected to bioassay using jar test method. The results were compared to those of a susceptible laboratory strain. Biochemical assays of esterases, monooxigenases and glutathione S-transferase (GST) levels were undertaken on German cockroaches from Imam and Bouali Cina Hospitals and the results were compared to a susceptible laboratory strain. RESULTS: The LT50 values of the three strains were 20.24+/-2.2, 19.87+/-2.3 and 8.89+/-0.26 for permethrin; 19.3+/-3.05, 17.6+/-0.68 and 8.8+/-0.99 for deltamethrin; 19.64+/-2.9, 18.66+/-3.45 and 8.64+/-0.62 min for cypermethrin, respectively. The mean -esterase activity of Imam and Bouali Cina Hospitals and susceptible strains were 6.941x10(-4), 6.940x10(-4) and 8.01x10(-5) nmol/min/mg protein; the mean -esterase activity in those strains were 5.8x10(-4), 4.25x10(-4) and 7.28x10(-5) nmol/min/mg protein; the mean content of p450 in the above-mentioned strains were 5.64x10(-6), 1.89x10(-6) and 1.2x10(-6) nmol/mg protein; the mean GST activity were 6.66x10(-2), 0.102 and 5.72x10(-2) mol/min/mg protein, respectively. CONCLUSION: The LT50 values and also the mean activity of all enzyme groups in field strains were significantly different from those of the susceptible strain, indicating a vigour tolerance to insecticides and pyrethroids in particular. Hence, insecticide resistance monitoring techniques should be put in place and also resistance management strategies and measures should be considered implementing in the area. PMID- 18392088 TI - Nitric oxide level in seminal plasma and its relation with sperm DNA damages. AB - BACKGROUND: Evidence supports the involvement of nitric oxide (NO) in a variety of male reproductive processes such as spermatogenesis, spermiogenesis, sperm motion, sperm metabolism and sperm capacitation. However, low concentration of NO is essential in biology and physiology of spermatozoa, but high amounts of NO is toxic and has negative effects on sperm functions. On the other hand, it is established that high amounts of NO have detrimental effects on DNA. The integrity of sperm DNA is an important factor in successful fertility and embryo development. It is hypothesized that supra physiological concentrations of NO in seminal plasma cause sperm DNA damage. The aim of this study was to determine sperm DNA damage by comet assay and its correlation with NO level in seminal plasma of fertile and infertile men. METHODS: Semen samples were collected from 45 patients and 70 healthy donors. The stable metabolites of NO (nitrite and nitrate) in seminal plasma were measured by Griess assay and DNA damage was determined using single cell gel electrophoresis (comet) assay method. RESULTS: The NO concentration in the seminal plasma of infertile males was significantly higher than fertile males (5.74+/-1.01 microM/L vs. 3.88+/-0.53 microM/L). There was a significant positive correlation between the NO concentration and sperm DNA comet value in infertile males (P<0.01, R=0.598). CONCLUSION: These results indicate that the overproduction of NO in genital tract of infertile males has a potential pathogenetic role in the reduction of sperm DNA integrity. PMID- 18392089 TI - Induction of apoptosis on K562 cell line and double strand breaks on colon cancer cell line expressing high affinity receptor for granulocyte macrophage-colony stimulating factor (GM-CSF). AB - BACKGROUND: Immunotoxins are comprised of both the cell targeting and the cell killing moieties. We previously established a new immunotoxin, i.e. Shiga toxin granulocyte macrophage-colony stimulating factor (StxA1-GM-CSF), comprises of catalytic domain of Stx, as a killing moiety and GM-CSF, as a cell targeting moiety. In this study, the ability of the immunotoxin to induce apoptosis and double strand breaks (DSB) on different cell lines was investigated. METHODS: The recombinant hybrid protein was expressed in bacterial expression system and purified with nickel-nitrilotriacetate acid resin. The K562 (erythroid leukemia) cell line and LS174 (colon carcinoma) were used in this study. The neutral comet assay was carried out for the detection of DSB and Hoechst staining was performed for apoptosis. RESULTS: StxA1-GM-CSF effectively induced apoptosis on K562 cell line and DNA Double Strand Break (DSB) were observed on colon cancer cell line treated with StxA1-GM-CSF. CONCLUSION: This novel action i.e. DNA damage might be a relevant mechanism of action for StxA1-GM-CSF that is designed to act as immunotoxin, although further investigation is required. PMID- 18392090 TI - Evaluation of growth inhibitory and apoptosis inducing activity of human calprotectin on the human gastric cell line (AGS). AB - BACKGROUND: Calprotectin is cytotoxic agent that its anti-tumor effects are governed through suppression of topoisomerase II; a key enzyme in apoptosis. In previous studies, cytotoxicity and apoptotic effects of calprotectin are shown on different cancer cell lines, but not human gastric cancer cell lines. In the present study, cytotoxicity and apoptotic effects of calprotectin on human gastric adenocarcinoma cancer cell line (AGS) were evaluated. METHODS: The AGS cells were exposed to the different concentrations of calprotectin for 24, 48 and 72 hours. Cell proliferation was assessed using dimethylthiazol diphenyl tetrazolium bromide assay and dye exclusion tests. For evaluation of cytotoxic mechanism in calprotectin on AGS cells, flow cytometric analysis was performed. RESULTS: Our results revealed that calprotectin induces growth inhibition of AGS in a dose- and time-dependent manner. Results of this investigation showed that sensitivity of AGS cells to cytotoxic effect of human calprotectin was highly remarkable. In addition, growth inhibitory effect of this cytotoxic agent mostly was governed through induction of apoptosis in the AGS cells. CONCLUSION: These findings indicated that calprotectin induces growth inhibition and apoptosis in the AGS cells. PMID- 18392091 TI - Rapid screening of toxigenic vibrio cholerae O1 strains from south Iran by PCR ELISA. AB - BACKGROUND: The ability to sensitively detect Vibrio cholera with PCR-ELISA method represents a considerable advancement over alternative more time-consuming methods for detection of this pathogen. The aim of this research is to evaluate the suitability of a PCR-enzyme-linked immunosorbent assay for sensitive and rapid detection of V. cholera O1. METHODS: The 398-bp sequence of a gene that codes for the cholera toxin B subunit was amplified by PCR. The digoxigenin labeled amplified products were coated on microplates and detected by ELISA. The PCR product was also hybridized with biotin labelled probe and detected by ELISA using streptavidin. RESULTS AND CONCLUSION: The specificity of the PCR was determined using 10 bacterial strains and 50 samples from south Iran. The detection limit was 0.5 pg of the genomic DNA and five bacterial cells. Adaptation of PCR into PCR-ELISA assay format facilitates specific and sensitive detection and diagnosis of human cholera disease. We conclude that this PCR-ELISA is a diagnostic method that specifically detects toxin genes in V. cholera O1 strains. It is more rapid and less cumbersome than other diagnostic methods for detection of toxicity in these strains. PMID- 18392092 TI - The full length hepatitis C virus polyprotein and interactions with the interferon-beta signalling pathways in vitro. AB - BACKGROUND: Hepatitis C is a global health problem. The exact mechanisms by which hepatitis C virus (HCV) can evade the host immune system have become controversial. Whether HCV polyproteins modulate IFN signalling pathways or HCV proteins are responsible for such a property is the subject of interest. Therefore, an efficient baculovirus delivery system was developed to introduce the whole genome of HCV1B minus 3'untranslated region (UTR) (HCV1BDelta3'UTR) into hepatoma cells. METHODS: The whole genome of HCV genotype 1b was developed into hepatoma cells. Also, two replicon constructs were used in this research: a recombinant baculovirus containing the culture adapted sub-genomic replicon (Fk5.1) derived from HCV genotype 1b, and a mutant form containing an inactivating mutation within the non-structural protein 5B (NS5B). RESULTS: As expected, the baculovirus carrying the FK5.1 replicon induced the production of IFN-beta as judged by the use of an IFN-beta promoter luciferase reporter construct, whereas the GND baculovirus (a control polymerase knock-out replicon) and the full-length 3'UTR deletant failed to induce luciferase expression. The activation of both IFN regulatory factor 3 (IRF3) and nuclear factor kappaB (NFkappaB), two transcription factors induced by dsRNA signalling were examined. Both the wild type and GND-mutant replicon blocked the dsRNA-induced activation of IRF3 and NFkappaB. CONCLUSION: Inhibition of the transcriptional response to IRF3 and NFkappaB seems to be one of the multiple mechanisms which HCV employs to escape the host immune defense. In contrast, the full length 3'UTR deletant had no significant effect on either transcription factor. These results may be attributed to the function of HCV subgenomic replicons when compared with full length 3'UTR deletant. PMID- 18392093 TI - The morphological changes of adult mouse testes after 60Co gamma-Radiation. AB - BACKGROUND: Cytotoxic therapy can lead to prolonged azoospermia or even sterility. In the present study, we investigated the morphological changes of mouse testes after gamma-Radiation. METHODS: After anaesthetizing of NMRI mice, testes and their surrounding tissues were irradiated using a cobalt therapy machine. Four experimental groups were irradiated with fractionated doses of: 1.5 + 8, 1.5 + 12 and 1.5 + 16 Gy (with an interval of 24 h) and single dose of 14 Gy. Non-irradiated mice were considered as control group. Testes were removed 4, 6 and 8 weeks following irradiation, weighed and processed for light microscopic study. Diameters of seminiferous tubules and their lumens, epithelium thickness, percentage of different types of tubules and number of spermatogenic cell were measured. Moreover, sperm count motility and viability rates were evaluated in epididymis. RESULTS: Number of normal tubules, epithelium thickness, tubules diameter and lumen diameter were significantly reduced with high dose irradiation in comparison with control testes. The recovery was observed after 8 weeks. Epididymal sperm count, motility and viability rates were significantly decreased in the irradiated mice comparing non-irradiated ones. These parameters were increased after 8 weeks. CONCLUSION: According to the results, irradiation can cause temporary azoospermia in mouse and this effect is reversible after 8 weeks. PMID- 18392094 TI - The assessment of feto-maternal hemorrhage in an artificial model using anti-D and anti-fetal hemoglobin antibody by flow cytometry. AB - BACKGROUND: When fetal red cells enter the maternal circulation from placenta, an event would be happened that is described as feto-maternal hemorrhage (FMH). This life-threatening condition could be detected by using RBC antigens (surface antigens and intracellular antigens). Therefore, the measurement of fetal RBC in an artificial model would be useful to calculate FMH and consequently the dosage of Rhogam for prophylaxis. The aim of the present study was to evaluate FMH in an artificial mixture model. METHODS: A series of 40 artificial specimens were prepared consisting of Rh(D) negative adult blood (non-immunized) spiked with varying amounts of Rh(D) positive cord blood from mothers between 20-30 years old in Shahid Beheshti Hospital, Tehran, Iran. Monoclonal anti-D and anti-HbF (fetal hemoglobin) were used for detection of fetal RBC in artificial mixture sample modeling. RESULTS: This study showed that the percentage of fetal cells in artificial sample for anti-D antigen is in ranges of 0.28%-0.32% for a 0.25% dilution mixture, and 1.3%-2.05% for the mixture with dilution 2%. In addition, the ranges of data for anti-HbF staining was obtained 0.2%-0.34% for the 0.25% dilution sample, and the ranges of 1.04-1.8% for the 2% dilution. The regression analysis indicated that the correlation of anti-D assessment with expected standard method was r2 = 0.9672 and anti-HbF assessment was r2 = 0.8842. CONCLUSION: Although both molecule targets could be used for detection of fetal RBC, in this model, anti-D staining was more accurate than anti-HbF staining. However, since anti-D can not be utilized for low-density or weak phenotype and other incompatibility, the anti-HbF labeling could be used for all FMH. PMID- 18392095 TI - The frequency of eight common point mutations in CYP21 gene in Iranian patients with congenital adrenal hyperplasia. AB - BACKGROUND: Congenital Adrenal Hyperplasia (CAH, the inherited inability to synthesize cortisol) is one of the most common (1 in 10000 to 1 in 15000) autosomal recessive disorders. More than 95% of cases of CAH are caused by 21 hydroxylase deficiency (21-OHD). Females with severe, classic 21-OHD are exposed to excess androgens prenatally and are born with virilized external genitalia. Most patients cannot synthesize sufficient aldosterone to maintain sodium balance and may develop potentially fatal salt wasting crisis if not treated. METHODS: We applied allele specific PCR to detect the eight common mutations in the CYP21 gene in patients. Fifty unrelated patients with symptoms of classical CAH were studied. RESULTS AND CONCLUSION: Seventy percent of our subjects had these mutations. The most frequent mutations were found to be I2G and del-8 bp (28% and 13%, respectively). The frequencies of other alleles were as following: I172N, 9%; V281L, 3%; exon 6 cluster (I236N, V237E and M239K), 4%; Q318X, 9%; R356W, 5%; and P30L, 0%. The frequency of mutations did not differ substantially from other ethnics, however, a higher rate of del-8 bp (13%) was found in our population. The aim of this study was to detect common mutations for setting up a molecular method for prenatal diagnosis. PMID- 18392096 TI - Molecular characterization of a Squalene epoxidase gene in dermatophyte pathogen Trichophyton tonsurans. AB - BACKGROUND: Trichophyton tonsurans is one of the dermatophyte fungi which invades the skin and hair of human. Several properties of this fungus have been investigated so far. However a few studies were carried out in the field of molecular biology of this fungus. In the present study, we tried to identify the Squalene epoxidase gene which is related to synthesis of ergosterol in this fungus. METHODS: Pairs of 23 and 24 nucleotides primers were designed from highly conserved regions of the similar genes in other fungi. Mentioned primers were utilized in PCR by using isolated genomic DNA of T. tonsurans whereas the PCR fragments were then sequenced. RESULTS AND CONCLUSION: Nucleotides (n = 558) have been sequenced from this new gene which encodes a polypeptide with 186 amino acids. Sequences comparison in gene data banks (NCBI, NIH) for this part of DNA and its deduced amino acid revealed significant homology with members of the eukaryotic Squalene epoxidase. PMID- 18392097 TI - Focus: issues in developmental biology education. PMID- 18392098 TI - A Mechanistic Approach for Modulation of Arsenic Toxicity in Human Lymphocytes by Curcumin, an Active Constituent of Medicinal Herb Curcuma longa Linn. AB - Chronic exposure of humans to high concentrations of arsenic in drinking water is associated with skin lesions, peripheral vascular disease, hypertension, blackfoot disease and a high risk of cancer. Arsenic induces single strand breaks, DNA-protein crosslinks and apurinic sites in DNA, which are prerequisites for induction of cancer. Amelioration of such damages with natural compounds could be an effective strategy to combat arsenic toxicity. Curcumin is the active ingredient of turmeric, a common household spice, which is a rich source of polyphenols and this compound has been extensively studied as a chemopreventive agent against many types of cancer. The present study investigates whether curcumin could counteract the DNA damage caused by arsenic as assessed by single cell gel electrophoresis (SCGE) using peripheral blood lymphocytes, from healthy donors. It was observed that DNA damage induced by arsenic could be efficiently reduced by curcumin and the effect was more pronounced when lymphocytes were pre incubated with curcumin prior to arsenic insult. Arsenic caused DNA damage by generation of reactive oxygen species (ROS) and enhancement of lipid peroxidation levels. Curcumin counteracted the damage by quenching ROS, decreasing the level of lipid peroxidation and increasing the level of phase II detoxification enzymes like catalase, superoxide dismutase and glutathione peroxidase. Curcumin also enhanced the DNA repair activity against arsenic induced damage. The expression of polymerase, a repair enzyme, was found to be highly elevated when arsenite induced damaged cells were allowed to repair in presence of curcumin. Results indicate that curcumin has significant role in confronting the deleterious effect caused by arsenic, which could be an economic mode of arsenic mitigation among rural population in West Bengal, India. PMID- 18392099 TI - Involvement of endoplasmic reticulum stress in myocardial apoptosis of streptozocin-induced diabetic rats. AB - Apoptosis plays critical role in diabetic cardiomyopathy and endoplasmic reticulum stress (ERS) is one of intrinsic apoptosis pathways. For previous studies have shown that endoplasmic reticulum become swell in diabetic myocardium and ERS was involved in diabetes mellitus and heart failure, this study aimed to demonstrate whether ERS was induced in myocardium of streptozocin (STZ)-induced diabetic rats. We established type 1 diabetic rat model with STZ intraperitoneal injection, used echocardiographic evaluation, hematoxylin-eosin staining and the terminal deoxynucleotidyl transferase-mediated DNA nick-end labeling staining to identify the existence of diabetic cardiomyopathy and enhanced apoptosis in the diabetic heart. We performed immunohistochemistry, Western blot and real time PCR to analysis two hallmarks of ERS, glucose regulated protein78 (Grp78) and Caspase12. We found both Grp78 and Caspase12 had enhanced expression in protein and mRNA levels in diabetic myocardium than normal rat's, and Caspase12 was activated in diabetic heart. Those results suggested that ERS was induced in STZ induced diabetic rats' myocardium, and ERS-associated apoptosis took part in the pathophysiology of diabetic cardiomyopathy. PMID- 18392100 TI - Neutrophil-dependent oxidative stress in ulcerative colitis. AB - Neutrophil accumulation within epithelial crypts and in the intestinal mucosa directly correlates with clinical disease activity and epithelial injury in ulcerative colitis (UC). Current advances have defined the mechanisms by which neutrophils are activated or migrate across mucosal epithelia. A better understanding of this process will likely provide new insights into novel treatment strategies for UC. Especially, activated neutrophils produce reactive oxygen and nitrogen species within intestinal mucosa, which induce oxidative stress. In clinically, we have succeeded to develop a novel granulocytes adsorptive apheresis therapy for UC. In this article, we discuss current advances to define the role of neutrophils-dependent oxidative stress in UC. PMID- 18392101 TI - Serum cardiac markers in patients with acute myocardial infarction: oxidative stress, C-reactive protein and N-terminal probrain natriuretic Peptide. AB - The aim of this study was to investigate the predictive value of an oxidative stress, C-reactive protein (CRP) and N-terminal probrain natriuretic peptide (NT proBNP) biomarkers in acute myocardial infarction (AMI). The study population contained 100 patients with AMI and 40 healthy subjects. Malondialdehyde (MDA) was measured as thiobarbituric acid reactive substances. Total antioxidant status (TAC) was assayed with colorimetric method. CRP and NT-proBNP was quantitated by immunoassay. MDA, CRP and NT-proBNP levels were found significantly high in patients with AMI as compared to healthy controls (p<0.01). Patients were divided into six groups based on the presence of disease history before AMI. In patients with non-disease before AMI. MDA, CRP and NT-proBNP levels were lowest among the patient groups. MDA levels in patients with hyperlipidemia/ diabetes/renal disease were higher than the other groups. TAC levels in patients with hypertension were lower than as compared to healthy controls (p<0.05). CRP levels in hypertension + hyperlipidemia patients and NT-proBNP levels in cardiovascular + hypertension patients were found high as compared to other patient groups. It is concluded that serum levels of MDA, CRP and NT-proBNP were significantly increased in patients with AMI and these markers were strongly predictive in AMI. PMID- 18392102 TI - Various Molecular Species of Diacylglycerol Hydroperoxide Activate Human Neutrophils via PKC Activation. AB - We have proposed that diacylglycerol hydroperoxide-induced unregulated signal transduction causes oxidative stress-related diseases. In this study, we investigated which molecular species of diacylglycerol hydroperoxide activated human peripheral neutrophils. All diacylglycerol hydroperoxides, diacylglycerol hydroxides, and diacyglycerols tested in the present study induced superoxide production by neutrophils. The ability to activate neutrophils among molecular species containing the same fatty acid composition was as follows; diacylglycerol hydroperoxide>diacylglycerol hydroxide>/=diacylglycerol. The diacylglycerol hydroperoxide composed of linoleate was a stronger activator for neutrophils than that composed of arachidonate. 1-Palmitoyl-2-linoleoylglycerol hydroperoxide (PLG OOH) was the strongest stimulator for neutrophils. We reconfirmed that PLG-OOH activated protein kinase C (PKC) in neutrophils. PLG-OOH induced the phosphorylation of p47(phox), a substrate of PKC and a cytosolic component of NADPH oxidase, in neutrophils, as did N-formyl-methionyl-leucyl-phenylalanine or 4beta-phorbol-12beta-myristate-13alpha-acetate. Moreover, the time course of p47(phox) phosphorylation was comparable to that of superoxide production. These results suggest that PLG-OOH activated intracellular protein kinase C. PLG-OOH, produced via an uncontrolled process, can act as a biological second messenger to cause inflammatory disease from oxidative stress. PMID- 18392103 TI - Indian herbal medicines: possible potent therapeutic agents for rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is an autoimmune disease of unknown etiology and is mainly characterized by the progressive erosion of cartilage leading to chronic polyarthritis and joint distortion. Although the exact pathogenesis of the disease has yet not been elucidated, however, studies suggest that cellular proliferation of synoviocytes result in pannus formation which damages the cartilage and bone. Recent reports also support the role of free radicals in its pathogenesis. Apart from the conventional treatment strategies using nonsteroidal anti-inflammatory drugs, disease modifying antirheumatic drugs and glucocorticoids, newer and safer drugs are continuously being searched, as long term usage of these drugs have resulted in adverse effects. Alternative medicine provides another approach for treatment of RA and currently a number of medicinal plants are under scientific evaluation to develop a novel drug. There is a dire need to investigate the complete therapeutic potential and adverse effects, if any, of these herbals for providing newer and safer treatment options with minimum side effects. In this review we have tried to explore various Indian ancient Ayurvedic, Unani and Tibbi, as also some Chinese and Korean, herbals for their potential to treat RA. PMID- 18392104 TI - The effect of acetaminophen on oxidative modification of low-density lipoproteins in hypercholesterolemic rabbits. AB - Oxidative modification of low-density lipoproteins (LDL) contributes to the pathology of atherosclerosis. Antioxidants may protect LDL against oxidative modification. Acetaminophen, a widely used analgesic and antipyretic agent, has significant antioxidant properties. However, there is little evidence to suggest that acetaminophen acts as an antioxidant for LDL oxidation in vivo. In this study, we investigated the in vivo effect of acetaminophen on LDL oxidation in hypercholesterolemic rabbits. The oxidative modification of LDL was identified by conjugated dienes and thiobarbituric acid-reactive substances (TBARS). In the cholesterol group which rabbits were fed a diet contained 1% g cholesterol for 8 weeks, TBARS contents and conjugated diene levels in the plasma and isolated LDL samples significantly increased compared with the control rabbits (p<0.05). However, in the cholesterol + acetaminophen group, the TBARS contents and conjugated diene levels were significantly lower than that of the cholesterol group (p<0.05). The results from in vitro studies also demonstrated that the LDL isolated from serum was oxidized by Cu(++) ions and this oxidation reduced in the presence of acetaminophen. The reduced oxidative modification of LDL by acetaminophen may be of therapeutic value in preventing the development and progression of atherosclerosis. PMID- 18392105 TI - Effects of green tea fractions on oxygen-induced retinal neovascularization in the neonatal rat. AB - This study aimed to investigate the preventive effects of green tea fractions (GTFs) on rat model of oxygen-induced retinopathy (OIR). Neonatal Sprague-Dawley rats were exposed to daily cycles of 80% oxygen (20.5 h), ambient air (0.5 h), and progressive return to 80% oxygen (3 h) until postnatal day 12 (P12), then the rats were placed in ambient air until P18. The green tea was fractionated by DM A50, DM-W, M-B, and M-W. The rats were treated once daily from P6 to P17 by gastric gavage of GTFs (0.05 or 0.01 g/ml) or distilled water (DW) at 50 microl/10 g body weight. On P18, the rats were sacrificed and the retinal samples were collected. The retinal neovascularization (NV) was scored and avascular areas (AVAs) were measured as a % of total retinal area (%AVAs) in ADPase stained retinas. The NV scores in 0.01 g/ml M-W were significantly lower than those in DW. The %AVAs in 0.05 g/ml DM-A50 and in 0.05 g/ml and 0.01 g/ml M-W were significantly lower than those in DW. There were less catechins, and less caffeine in M-W fraction compared with other GTFs, suggesting components of green tea except for catechins and caffeine might suppress the neovascularization in rat model of OIR. PMID- 18392106 TI - Current status of herbal drugs in India: an overview. AB - Herbal drugs constitute a major share of all the officially recognised systems of health in India viz. Ayurveda, Yoga, Unani, Siddha, Homeopathy and Naturopathy, except Allopathy. More than 70% of India's 1.1 billion population still use these non-allopathic systems of medicine. Currently, there is no separate category of herbal drugs or dietary supplements, as per the Indian Drugs Act. However, there is a vast experiential-evidence base for many of the natural drugs. This offers immense opportunities for Observational Therapeutics and Reverse Pharmacology. Evidence-based herbals are widely used in the diverse systems and manufactured, as per the pharmacopoeial guidelines, by a well-organised industry. Significant basic and clinical research has been carried out on the medicinal plants and their formulations, with the state-of-the-art methods in a number of Institutes/Universities. There are some good examples. Indian medicinal plants also provide a rich source for antioxidants that are known to prevent/delay different diseased states. The antioxidant protection is observed at different levels. The medicinal plants also contain other beneficial compounds like ingredients for functional foods. Hence, the global knowledge about Ayurveda and Indian herbals will hopefully be enhanced by information on the evidence-base of these plants. This will yield rich dividends in the coming years. PMID- 18392107 TI - The amygdala and the experience of affect. AB - The current study examined the hypothesis that amygdala activation serves as a neural precondition for negative affective experience. Participants' affective experience was measured by asking them to report on their momentary experiences several times a day over the course of a month using an electronic experience sampling procedure. One year later, participants viewed backwardly masked depictions of fear while functional magnetic resonance imaging was used to measure their amygdala and fusiform gyrus activation. Negative affect, as measured during the experience-sampling procedure 1-year prior, was positively correlated with amygdala activation in response to these brief presentations of fear depictions. Furthermore, descriptive analyses indicated that fusiform gyrus activation and negative affective experience in the scanner were associated for participants reporting increased nervousness during the imaging procedure. The results are consistent with the interpretation that the amygdala contributes to negative affective experience by increasing perceptual sensitivity for negative stimuli. PMID- 18392108 TI - 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP)-formulated, immune-stimulatory vascular endothelial growth factor a small interfering RNA (siRNA) increases antitumoral efficacy in murine orthotopic hepatocellular carcinoma with liver fibrosis. AB - Most experimental therapy studies are performed in mice that bear subcutaneous or orthotopic hepatoma but are otherwise healthy and nonfibrotic. The majority of hepatocellular carcinoma (HCC), however, develops in patients suffering from preexisting liver fibrosis. We investigated the efficacy of a standard experimental therapeutic approach to interrupt the vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR) cascade via VEGF-A silencing, with or without 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP; cationic lipid) formulation, in HCC mice with preexisting liver fibrosis. The data show that intraperitoneal treatment with naked VEGF-A small interfering RNA (siRNA; 200 microg/kg) was inefficient to treat HCC implanted into fibrotic livers. VEGF-A siRNA containing an immunostimulatory motif in combination with DOTAP formulation significantly reduced hepatic VEGF-A expression and additionally activated the innate and adapted immune system as shown by an increased intrahepatic interferon type 1 response (68-fold increased beta-interferon expression). DOTAP-formulated VEGF-A siRNA markedly improved VEGF-A siRNA uptake and enhanced the antitumor response. This study shows for the first time the therapeutic feasibility of using synergistic effects (gene silencing and activation of the immune system) united in one siRNA sequence to reduce HCC growth and metastasis in mice with preexisting liver fibrosis. We expect that these results will help to direct and improve future experimental gene-silencing approaches and establish more efficient antitumoral therapies against HCC. PMID- 18392109 TI - The HapMap Resource is Providing New Insights into Ourselves and its Application to Pharmacogenomics. AB - The exploration of quantitative variation in complex traits such as gene expression and drug response in human populations has become one of the major priorities for medical genetics. The International HapMap Project provides a key resource of genotypic data on human lymphoblastoid cell lines derived from four major world populations of European, African, Chinese and Japanese ancestry for researchers to associate with various phenotypic data to find genes affecting health, disease and response to drugs. Recent progress in dissecting genetic contribution to natural variation in gene expression within and among human populations and variation in drug response are two examples in which researchers have utilized the HapMap resource. The HapMap Project provides new insights into the human genome and has applicability to pharmacogenomics studies leading to personalized medicine. PMID- 18392110 TI - Latency associated peptide has in vitro and in vivo immune effects independent of TGF-beta1. AB - Latency Associated Peptide (LAP) binds TGF-beta1, forming a latent complex. Currently, LAP is presumed to function only as a sequestering agent for active TGF-beta1. Previous work shows that LAP can induce epithelial cell migration, but effects on leukocytes have not been reported. Because of the multiplicity of immunologic processes in which TGF-beta1 plays a role, we hypothesized that LAP could function independently to modulate immune responses. In separate experiments we found that LAP promoted chemotaxis of human monocytes and blocked inflammation in vivo in a murine model of the delayed-type hypersensitivity response (DTHR). These effects did not involve TGF-beta1 activity. Further studies revealed that disruption of specific LAP-thrombospondin-1 (TSP-1) interactions prevented LAP-induced responses. The effect of LAP on DTH inhibition depended on IL-10. These data support a novel role for LAP in regulating monocyte trafficking and immune modulation. PMID- 18392111 TI - Electrical characterization of protein molecules by a solid-state nanopore. AB - The authors measured ionic current blockages caused by protein translocation through voltage-biased silicon nitride nanopores in ionic solution. By calculating the mean amplitude, time duration, and the integral of current blockages, they estimated the relative charge and size of protein molecules at a single molecule level. The authors measured the change in protein charge of bovine serum albumin (BSA) protein induced by pH variation. They also confirmed that BSA molecules indeed traverse nanopores using an improved chemiluminescent analysis. They demonstrated that a larger protein fibrinogen could be distinguished from BSA by a solid-state nanopore measurement. PMID- 18392112 TI - Song matching, overlapping, and switching in the banded wren: the sender's perspective. AB - Interpreting receiver responses to on-territory playback of aggressive signals is problematic. One solution is to combine such receiver-perspective experiments with a sender-perspective experiment that allows subjects to demonstrate how their choice of singing strategies is associated with their approach behavior. Here we report the results of a sender-perspective study on the banded wren (Thryothorus pleurostictus), and combine information on context and results of previous receiver-perspective experiments to clarify function. Territorial males were presented with a 5-min playback consisting of song types present in their repertoire. We assessed the degree to which the subjects' song matching rate, overlapping rate, and song-type versatility were correlated with their approach latency, closeness of approach, latency to first retreat, and time spent close to the speaker. Male age, breeding stage, and features of the playback stimuli were also considered. Song matching was associated with rapid and close approach, consistent with the receiver-perspective interpretation of type matching as a conventional signal of aggressive motivation. Overlapping was associated with earlier retreat, and together with the aversive receiver response to our previous overlapping playback experiment suggests that overlapping is a defensive withdrawal signal. High versatility was associated with slower first retreat from the speaker and high levels of reciprocal matching between subject and playback. Males with fledglings sang with particularly low versatility and approached the speaker aggressively, whereas males with nestlings overlapped more and retreated quickly. Finally, older males matched more but overlapped less. PMID- 18392114 TI - Brain SPECT Simulation Using Half-Cone-Beam Collimation and Single-Revolution Helical-Path Acquisition. AB - In this study related to human brain SPECT imaging, simulation of half-cone-beam collimation and helical-path data acquisition is performed. We discuss problems related to circular-orbit acquisition using cone-beam collimation, such as shoulder interference resulting in object truncation, and insufficient sampling of the object resulting in axial distortions in the reconstructed images. We demonstrate that a triple-camera SPECT system with half-cone-beam collimation and single-revolution helical-path acquisition eliminates both issues and offers substantially improved sampling and almost artifact-free reconstruction of the object. PMID- 18392113 TI - Prefrontal cortex damage abolishes brand-cued changes in cola preference. AB - Human decision-making is remarkably susceptible to commercial advertising, yet the neurobiological basis of this phenomenon remains largely unexplored. With a series of Coke and Pepsi taste tests we show that patients with damage specifically involving ventromedial prefrontal cortex (VMPC), an area important for emotion, did not demonstrate the normal preference bias when exposed to brand information. Both comparison groups (neurologically normal adults and lesion patients with intact VMPC) preferred Pepsi in a blind taste test, but in subsequent taste tests that featured brand information ('semi-blind' taste tests), both comparison groups' preferences were skewed toward Coke, illustrating the so-called 'Pepsi paradox'. Like comparison groups, the VMPC patients preferred Pepsi in the blind taste test, but unlike comparison groups, the VMPC patients maintained their Pepsi preference in the semi-blind test. The result that VMPC damage abolishes the 'Pepsi paradox' suggests that the VMPC is an important part of the neural substrate for translating commercial images into brand preferences. PMID- 18392115 TI - Eleven microsatellite loci isolated from the banded wren (Thryothorus pleurostictus). AB - We describe 11 microsatellite loci isolated from the Banded Wren (Thryothorus pleurostictus), a Neotropical species for which understanding the genetic mating system is important for testing questions about the species' unusual vocal behavior. Screening of these loci revealed extremely low allelic variation in a Costa Rican population. Allelic variation at these and other previously developed loci is substantially higher in two other wren species, the southern house wren (Troglodytes aedon bonariae) and rufous-and-white wren (Thryothorus rufalbus), suggesting that the low allelic diversity in the banded wren results from demographic bottlenecks rather than locus-sampling artifacts. PMID- 18392116 TI - Understanding Psychopathology: Melding Behavior Genetics, Personality, and Quantitative Psychology to Develop an Empirically Based Model. AB - Research on psychopathology is at a historical crossroads. New technologies offer the promise of lasting advances in our understanding of the causes of human psychological suffering. Making the best use of these technologies, however, requires an empirically accurate model of psychopathology. Much current research is framed by the model of psychopathology portrayed in current versions of the Diagnostic and Statistical Manual of Mental Disorders (DSM; American Psychiatric Association, 2000). Although the modern DSMs have been fundamental in advancing psychopathology research, recent research also challenges some assumptions made in the DSM-for example, the assumption that all forms of psychopathology are well conceived of as discrete categories. Psychological science has a critical role to play in working through the implications of this research and the challenges it presents. In particular, behavior-genetic, personality, and quantitative psychological research perspectives can be melded to inform the development of an empirically based model of psychopathology that would constitute an evolution of the DSM. PMID- 18392117 TI - Genomic response of the nematode Caenorhabditis elegans to spaceflight. AB - On Earth, it is common to employ laboratory animals such as the nematode Caenorhabditis elegans to help understand human health concerns. Similar studies in Earth orbit should help understand and address the concerns associated with spaceflight. The "International Caenorhabditis elegans Experiment FIRST" (ICE FIRST), was carried out onboard the Dutch Taxiflight in April of 2004 by an international collaboration of laboratories in France, Canada, Japan and the United States. With the exception of a slight movement defect upon return to Earth, the result of altered muscle development, no significant abnormalities were detected in spaceflown C. elegans. Work from Japan revealed apoptosis proceeds normally and work from Canada revealed no significant increase in the rate of mutation. These results suggest that C. elegans can be used to study non lethal responses to spaceflight and can possibly be developed as a biological sensor. To further our understanding of C. elegans response to spaceflight, we examined the gene transcription response to the 10 days in space using a near full genome microarray analysis. The transcriptional response is consistent with the observed normal developmental timing, apoptosis, DNA repair, and altered muscle development. The genes identified as altered in response to spaceflight are enriched for genes known to be regulated, in C. elegans, in response to altered environmental conditions (Insulin and TGF-beta regulated). These results demonstrate C. elegans can be used to study the effects of altered gravity and suggest that C. elegans responds to spaceflight by altering the expression of at least some of the same metabolic genes that are altered in response to differing terrestrial environments. PMID- 18392118 TI - Longitudinal Studies With Outcome-Dependent Follow-up: Models and Bayesian Regression. AB - We propose Bayesian parametric and semiparametric partially linear regression methods to analyze the outcome-dependent follow-up data when the random time of a follow-up measurement of an individual depends on the history of both observed longitudinal outcomes and previous measurement times. We begin with the investigation of the simplifying assumptions of Lipsitz, Fitzmaurice, Ibrahim, Gelber, and Lipshultz, and present a new model for analyzing such data by allowing subject-specific correlations for the longitudinal response and by introducing a subject-specific latent variable to accommodate the association between the longitudinal measurements and the follow-up times. An extensive simulation study shows that our Bayesian partially linear regression method facilitates accurate estimation of the true regression line and the regression parameters. We illustrate our new methodology using data from a longitudinal observational study. PMID- 18392119 TI - Evaluation of Multiclass Model Observers in PET LROC Studies. AB - A localization ROC (LROC) study was conducted to evaluate nonprewhitening matched filter (NPW) and channelized NPW (CNPW) versions of a multiclass model observer as predictors of human tumor-detection performance with PET images. Target localization is explicitly performed by these model observers. Tumors were placed in the liver, lungs, and background soft tissue of a mathematical phantom, and the data simulation modeled a full-3D acquisition mode. Reconstructions were performed with the FORE+AWOSEM algorithm. The LROC study measured observer performance with 2D images consisting of either coronal, sagittal, or transverse views of the same set of cases. Versions of the CNPW observer based on two previously published difference-of-Gaussian channel models demonstrated good quantitative agreement with human observers. One interpretation of these results treats the CNPW observer as a channelized Hotelling observer with implicit internal noise. PMID- 18392120 TI - Verb inflections in agrammatic aphasia: Encoding of tense features. AB - Across most languages, verbs produced by agrammatic aphasic individuals are frequently marked by syntactically and semantically inappropriate inflectional affixes, such as Last night, I walking home. As per language production models, verb inflection errors in English agrammatism could arise from three potential sources: encoding the verbs' morphology based on temporal information at the conceptual level, accessing syntactic well-formedness constraints of verbal morphology, and encoding morphophonological form. We investigate these aspects of encoding verb inflections in agrammatic aphasia. Using three sentence completion experiments, it was demonstrated that production of verb inflections was impaired whenever temporal reference was involved; while morphological complexity and syntactic constraints were less likely to be the source of verb inflection errors in agrammatism. These findings are discussed in relation to current language production models. PMID- 18392121 TI - Proceedings of the 10th Asian Pacific Congress of Clinical Biochemistry in conjunction with the Australasian Association of Clinical Biochemists' 42nd Annual Scientific Conference. PMID- 18392122 TI - Laboratory turnaround time. AB - Turnaround time (TAT) is one of the most noticeable signs of laboratory service and is often used as a key performance indicator of laboratory performance. This review summarises the literature regarding laboratory TAT, focusing on the different definitions, measures, expectations, published data, associations with clinical outcomes and approaches to improve TAT. It aims to provide a consolidated source of benchmarking data useful to the laboratory in setting TAT goals and to encourage introduction of TAT monitoring for continuous quality improvement. A 90% completion time (sample registration to result reporting) of <60 minutes for common laboratory tests is suggested as an initial goal for acceptable TAT. PMID- 18392123 TI - HbA1c standardisation: history, science and politics. AB - Significant analytical improvements have occurred since glycated haemoglobin (GHb), measured as total HbA(1), was first used in routine clinical laboratories around 1977. Following the publication of the Diabetes Control and Complications Trial (DCCT) study in 1993 the issue of international standardisation became an important objective for scientists and clinicians. The lack of international standardisation led several countries to develop national standardisation programs. The International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) Working Group on Standardisation of HbA(1c) established a true international reference measurement system for HbA(1c) and the successful preparation of pure HbA(1c) calibration material that should lead to further improvements in inter-method and inter-laboratory variability. Reporting of HbA(1c) has been agreed using the units of mmol/mol (IFCC) and percent (National Glycohemoglobin Standardization Program, NGSP). PMID- 18392125 TI - Standardisation--the theory and the practice. PMID- 18392124 TI - Reference materials and commutability. AB - Maintaining accurate laboratory measurements over time is crucial for assuring appropriate patient care and disease management. Accurate results over time and location are achieved by standardising measurements and establishing traceability to a reference system. Reference materials are key components of such reference systems and for establishing traceability. Commutability of reference materials is a critical property to ensure they are fit for use. Commutability is defined as the equivalence of the mathematical relationships between the results of different measurement procedures for a reference material and for representative samples from healthy and diseased individuals. This material characteristic is of special importance for measurement procedures that are optimised for measuring analytes directly in patient samples. The commutability of a reference material is measurement procedure specific and its assessment requires special experimental designs. This review explains the importance of commutability and summarises different experimental approaches described in the literature that have been used to assess the commutability of reference materials in clinical chemistry. PMID- 18392126 TI - Traceability in clinical enzymology. AB - The primary goal of standardisation for measurements of catalytic concentrations of enzymes is to achieve comparable results in human samples, independent of the reagent kits, instruments and laboratory where the assay is carried out. In order to pursue this objective, the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) has established reference systems for the most important clinical enzymes. These systems are based on three requirements: a) reference measurement procedures that are extensively evaluated and carefully described; b) certified reference materials; and c) a network of reference laboratories operating in a highly controlled manner. Using these reference systems and the manufacturer's standing procedures, industry can assign traceable values to commercial calibrators. Clinical laboratories, which use routine procedures with validated calibrators to measure human specimens, can finally obtain values which are traceable to higher-order reference procedures. These reference systems constitute the structure of the traceability chain to which the routine methods can be linked via an appropriate calibration process, provided that they have a comparable specificity (i.e. they are measuring the same quantity). PMID- 18392127 TI - Reference materials and reference measurement procedures: an overview from a national metrology institute. AB - An outline of the processes involved in both certified clinical reference material production and clinical reference measurement procedure development at the National Institute of Standards and Technology (NIST), the national metrology institute of the United States, is presented. The role that NIST and other national metrology institutes play in the metrological traceability of certified reference material is discussed. Highlighted are the challenges associated with the development of reference measurement systems for complex clinical analytes, such as proteins, and examples of existing efforts in this area are given. Examples of recent international collaborations in developing certified reference materials for analytes such as cardiac troponin I, brain natriuretic peptide, and serum creatinine demonstrate the close cooperation that national metrology institutes must have with the clinical community to establish complete reference measurement systems. PMID- 18392128 TI - Standardisation of reference intervals: an Australasian view. AB - The development of regional databases and doctors' desktop programs that accept pathology results from different laboratories should improve patient care by allowing easy assessment of cumulative data. This has the potential to be unnecessarily confusing unless laboratories contributing to the databases provide standardised results and common reference intervals, where this is valid. The analytical methods that produce significantly different results need to be reported in a manner that avoids inappropriate interpretation. The process of setting reference intervals requires an organisational structure which enables appropriate intervals to be set taking all relevant factors into account, including the opinions of expert clinicians. There must also be criteria for analytical agreement between the laboratories involved based on comparison studies using patient samples.A network of QA groups across Australasia, with leadership from the AACB and RCPA, should be formed to share the ongoing work of defining reference intervals (RIs) for common tests, and reviewing them as the testing environment changes with the introduction of new techniques and instruments. PMID- 18392129 TI - Requirements for reference (calibration) laboratories in laboratory medicine. AB - In addition to reference measurement procedures and reference materials, reference or calibration laboratories play an integral role in the implementation of measurement traceability in routine laboratories. They provide results of measurements using higher-order methods, e.g. isotope dilution mass spectrometry and may assign values to materials to be used for external quality assessment programs and to secondary reference materials. The requirements for listing of laboratories that provide reference measurement services include a statement of the metrological level or principle of measurement, accreditation as a calibration laboratory according to ISO 15195 and the participation in a proficiency testing system (regular inter-laboratory comparisons) for reference laboratories. Ring trials are currently conducted for thirty well-defined measurands and the results are made available to all laboratories. Through the use of reference laboratory services that are listed by the Joint Committee for Traceability in Laboratory Medicine there is the opportunity to further promote traceability and standardisation of laboratory measurements. PMID- 18392130 TI - EQA from an Australian perspective. AB - Enrolment in external quality assurance programs is part of the accreditation process for medical laboratories in Australia, with the majority of Australian laboratories being enrolled in programs from RCPA Quality Assurance Programs Pty Limited, a company owned by the Royal College of Pathologists of Australasia. An important feature of these programs is that they have been developed with the involvement and contribution of the profession. For example, the Chemical Pathology programs are a joint venture between the company and the Australasian Association of Clinical Biochemists (AACB). Some of the unique features of the programs are the composition of the material, the use of target values, the structure and information in the reports and the use of the internet for data entry and data review. Over the past thirty years, the development of these programs has made a significant contribution to the quality of laboratories in Australia. PMID- 18392131 TI - A small-molecule furin inhibitor inhibits cancer cell motility and invasiveness. AB - Furin, a member the proprotein convertase (PC) family, processes inactive precursor proteins to functional proteins within the Golgi/trans-Golgi network secretory pathway. Furin and other PC family members (furin/PCs) activate proteins vital to proper physiological functioning, including growth factors and hormones, receptors, plasma proteins, and matrix metalloproteases (MMPs). Additionally, the expression and activity of furin/PC are necessary for processing substrates important for cell transformation and tumor progression, metastasis, and angiogenesis. Furin processing of the remodeling protease membrane type-1 matrix metalloproteinase (MT1-MMP) enhances cellular motility and invasiveness, contributing to aggression and metastatic potential cancer cells. Whereas overexpression and activity of furin/PC exacerbate the cancer phenotype, inhibition of its activity decreases or nullifies furin/PC-mediated effects, and thus, inhibition of furin may be a viable route to cancer therapy. Recently, we identified a small-molecule inhibitor of furin, named B3, by high-throughput screening with a K(i) and IC(50) of 12 microM. Here, we show that this cell permeable, small-molecule compound inhibits furin-mediated cleavage of proMT1 MMP, resulting in decreased MMP-2 activation and cell motility in CHO cells expressing proMT1-MMP. Additionally, this molecule inhibited proMT1-MMP processing, complete MMP-2 maturation, and invasiveness of human fibrosarcoma cells (HT1080). PMID- 18392132 TI - Tumor vascularity assessed by magnetic resonance imaging and intravital microscopy imaging. AB - Gadopentetate dimeglumine (Gd-DTPA)-based dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) is considered to be a useful method for characterizing the vascularity of tumors. However, detailed studies of experimental tumors comparing DCE-MRI-derived parametric images with images of the morphology and function of the microvascular network have not been reported. In this communication, we describe a novel MR-compatible mouse dorsal window chamber and report comparative DCE-MRI and intravital microscopy studies of A-07 GFP tumors xenografted to BALB/c nu/nu mice. Blood supply time (BST) images (i.e., images of the time from when arterial blood enters a tumor through the supplying artery until it reaches a vessel segment within the tumor) and morphologic images of the microvascular network were produced by intravital microscopy. Images of E.F (E is the initial extraction fraction of Gd-DTPA and F is perfusion) were produced by subjecting DCE-MRI series to Kety analysis. The E.F images mirrored the morphology (microvascular density) and the function (BST) of the microvascular networks well. Tumor regions showing high E.F values colocalized with tumor regions showing high microvascular density and low BST values. Significant correlations were found between E.F and microvascular density and between E.F and BST, both within and among tumors. PMID- 18392134 TI - Plasticity in tumor-promoting inflammation: impairment of macrophage recruitment evokes a compensatory neutrophil response. AB - Previous studies in the K14-HPV/E(2) mouse model of cervical carcinogenesis demonstrated that infiltrating macrophages are the major source of matrix metalloproteinase 9 (MMP-9), a metalloprotease important for tumor angiogenesis and progression. We observed increased expression of the macrophage chemoattractant, CCL2, and its receptor, CCR2, concomitant with macrophage influx and MMP-9 expression. To study the role of CCL2-CCR2 signaling in cervical tumorigenesis, we generated CCR2-deficient K14-HPV/E(2) mice. Cervixes of CCR2 null mice contained significantly fewer macrophages. Surprisingly, there was only a modest delay in time to progression from dysplasia to carcinoma in the CCR2 deficient mice, and no difference in end-stage tumor incidence or burden. Moreover, there was an unexpected persistence of MMP-9 activity, associated with increased abundance of MMP-9(+) neutrophils in tumors from CCR2-null mice. In vitro bioassays revealed that macrophages produce soluble factor(s) that can suppress neutrophil dynamics, as evidenced by reduced chemotaxis in response to CXCL8, and impaired invasion into three-dimensional tumor masses grown in vitro. Our data suggest a mechanism whereby CCL2 attracts proangiogenic CCR2(+) macrophages with the ancillary capability to limit infiltration by neutrophils. If such tumor-promoting macrophages are suppressed, MMP-9(+) neutrophils are then recruited, providing alternative paracrine support for tumor angiogenesis and progression. PMID- 18392135 TI - The growth factor Midkine antagonizes VEGF signaling in vitro and in vivo. AB - Midkine (MDK) is a heparin-binding growth factor involved in growth, survival, migration, and differentiation of various target cells and dysregulation of MDK signaling is implicated in a variety of inflammatory diseases and cancers. Although MDK has been reported to act on endothelial cells and to have proangiogenic effects, the exact role of MDK in angiogenesis is poorly defined. Here, we report that MDK is actually a modulator of angiogenesis and that it can abrogate the vascular endothelial growth factor A (VEGF-A)-induced proliferation of human microvascular endothelial cells in vitro through the downregulation of proangiogenic cytokines and through the upregulation of the antiangiogenic factor, tissue inhibitor of metalloproteinase 2. Phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR-2) and of downstream signaling molecules, such as phosphatidylinositol-3-kinase and mitogen-activated protein kinases, is also impaired. Moreover, MDK downregulates VEGF-A-induced neovascularization and vascular permeability in vivo. We propose a model in which MDK is a new modulator of the VEGF-A-VEGFR-2 axis. PMID- 18392133 TI - p21-Activated kinase 1 coordinates aberrant cell survival and pericellular proteolysis in a three-dimensional culture model for premalignant progression of human breast cancer. AB - Overexpression of p21-activated kinase 1 (PAK1) occurs during the progression of human breast cancer. We have investigated the role of PAK1 in the premalignant progression of the MCF10 series of human breast epithelial cell lines. Levels of PAK1 expression and activation increased with premalignant progression, and expression of dominant-negative (DN) PAK1 reduced both cell proliferation and migration/invasion. In three-dimensional (3D) overlay cultures in reconstituted basement membrane, the MCF10 series produced an in vitro model for premalignant progression. MCF10AneoT cells formed a hyperplastic morphology in which some spheroids developed abnormal lumens. The MCF10.AT1 line exhibited an atypical hyperplastic morphology of abnormal spheroid clusters that did not form lumens. The MCF10.DCIS cells exhibited dysplastic growth. Expression of DN-PAK1 promoted lumen formation in 3D-cultured MCF10A, NeoT, and AT1 structures, suggesting partial reversion of the premalignant phenotype, but did not affect the atypical budding of AT1 structures or the dysplastic growth of ductal carcinoma in situ structures. Aberrant proteolysis is another important characteristic of breast cancer progression and invasion. DN-PAK1 or knock-down of PAK1 reduced pericellular proteolysis of DQ-collagen IV in the 3D cultures. Treatment of cells with an inhibitor of Rac1 also reduced pericellular proteolysis, and this reduction was reversed by the expression of activated PAK1. Our conclusion is that overexpressed and activated PAK1 may be a key coordinator of aberrant cell survival and proteolysis in breast cancer progression. PMID- 18392136 TI - Glucosamine-bound near-infrared fluorescent probes with lysosomal specificity for breast tumor imaging. AB - Noninvasive imaging of lysosomes will be useful 1) to elucidate the role of lysosomal parameters in cancer, 2) to diagnose malignant lesions, and 3) to evaluate future lysosome-targeted anticancer therapies. Lysosome-specific labeling of glucosamine-bound near-infrared (NIR) fluorescent probes, IR-1 and IR 2, but not control probe IR-15 without the glucosamine moiety, was observed by fluorescence microscopy in human breast epithelial cell lines. Lysosome labeling and tumor specificity of these NIR probes were investigated by dynamic optical imaging and immunofluorescence staining in human breast tumor xenografts. IR-1 and IR-2 demonstrated faster lysosome labeling rates in highly aggressive MDA-MB 231 and MDA-MB-435 cells compared with less aggressive MCF-7 and nontumorigenic MCF-12A cells. IR-1 and IR-2, but not IR-15, accumulated in human MDA-MB-231, MDA MB-435, and MCF-7 breast tumor xenografts in vivo. IR-2 demonstrated the highest maximum fluorescence and tumor/normal tissue ratios in all tumor models. Specific lysosome labeling from IR-2 in vivo was validated by colocalization of the NIR fluorescence with CD63 immunofluorescence in tumor sections. IR-1 and IR-2 demonstrated high lysosome-labeling ability and breast tumor-targeting specificity in vitro and in vivo. They are promising for diagnosing malignant lesions and may provide a means for evaluating and monitoring future lysosome targeted anticancer therapies. PMID- 18392137 TI - Hypoxia increases breast cancer cell-induced lymphatic endothelial cell migration. AB - Because tumors are characterized by hypoxic environments, we used a novel in vitro noninvasive magnetic resonance imaging assay to examine the influence of invasive MDA-MB-231 breast cancer cells on the invasion and migration of human dermal lymphatic microvascular endothelial cells (HMVEC-dLy) under normoxic and hypoxic conditions. Nonmalignant immortalized MCF-12A human mammary epithelial cells instead of cancer cells or chambers with HMVEC-dLy alone were used as controls for comparison. HMVEC-dLy cells were labeled with a T(2) contrast agent (Feridex), and their invasion and migration through extracellular matrix under normoxic and hypoxic conditions were monitored using magnetic resonance imaging. A significant increase in the invasion and migration of HMVEC-dLy cells was detected in the presence of cancer cells, which further increased significantly under hypoxic conditions. HMVEC-dLy cells formed interconnecting strands extending toward the cancer cells under normoxic but not under hypoxic conditions. Following reoxygenation, these interconnecting strands, extending from HMVEC-dLy cells toward the cancer cells, were observed. These data demonstrate the importance of hypoxia in lymphatic endothelial cell invasion and migration through extracellular matrix in the presence of cancer cells. PMID- 18392138 TI - Utility of a herpes oncolytic virus for the detection of neural invasion by cancer. AB - Prostate, pancreatic, and head and neck carcinomas have a high propensity to invade nerves. Surgical resection is a treatment modality for these patients, but it may incur significant deficits. The development of an imaging method able to detect neural invasion (NI) by cancer cells may guide surgical resection and facilitate preservation of normal nerves. We describe an imaging method for the detection of NI using a herpes simplex virus, NV1066, carrying tyrosine kinase and enhanced green fluorescent protein (eGFP). Infection of pancreatic (MiaPaCa2), prostate (PC3 and DU145), and adenoid cystic carcinoma (ACC3) cell lines with NV1066 induced a high expression of eGFP in vitro. An in vivo murine model of NI was established by implanting tumors into the sciatic nerves of nude mice. Nerves were then injected with NV1066, and infection was confirmed by polymerase chain reaction. Positron emission tomography with [(18)F]-2'-fluoro-2' deoxyarabinofuranosyl-5-ethyluracil performed showed significantly higher uptake in NI than in control animals. Intraoperative fluorescent stereoscopic imaging revealed eGFP signal in NI treated with NV1066. These findings show that NV1066 may be an imaging method to enhance the detection of nerves infiltrated by cancer cells. This method may improve the diagnosis and treatment of patients with neurotrophic cancers by reducing injury to normal nerves and facilitating identification of infiltrated nerves requiring resection. PMID- 18392139 TI - Inhibiting colorectal carcinoma growth and metastasis by blocking the expression of VEGF using RNA interference. AB - Angiogenesis plays an essential role in tumor growth and metastasis and is a promising target for cancer therapy. Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis. The present study was designed to determine the role of VEGF in tumor growth and metastasis using RNA interference (RNAi) technology. Four small interfering RNA (siRNA) sequences for the VEGF gene were cloned into expression plasmids and transfected into human colorectal carcinoma (CRC) SW620 cells. Stable transfection of these plasmids decreased VEGF protein expression, leading to the potent suppression of tumor cell proliferation, migration, invasion, and angiogenesis in vitro. Furthermore, in subcutaneous and intrasplenic/portal injection models involving athymic nude mice, the tumor growth and metastasis of SW620 cells expressing VEGF siRNA were significantly inhibited compared with untransfected cells or cells transfected with control vector alone. Immunohistochemical analyses of tumor sections revealed a decreased vessel density and decreased VEGF expression in the animals where siRNA against VEGF were expressed. These results indicate that RNAi of VEGF can be an effective antiangiogenic strategy for CRC. PMID- 18392140 TI - Noninvasive magnetic resonance spectroscopic pharmacodynamic markers of a novel histone deacetylase inhibitor, LAQ824, in human colon carcinoma cells and xenografts. AB - The aim of this work was to use phosphorus magnetic resonance spectroscopy ((31)P MRS) to investigate the pharmacodynamic effects of LAQ824, a histone deacetylase (HDAC) inhibitor. Human HT29 colon carcinoma cells were examined by (31)P MRS after treatment with LAQ824 and another HDAC inhibitor, suberoylanilide hydroxamic acid. HT29 xenografts and tumor extracts were also examined using (31)P MRS, pre- and post-LAQ824 treatment. Histone H3 acetylation was determined using Western blot analysis, and tumor microvessel density by immunohistochemical staining of CD31. Phosphocholine showed a significant increase in HT29 cells after treatment with LAQ824 and suberoylanilide hydroxamic acid. In vivo, the ratio of phosphomonoester/total phosphorus (TotP) signal was significantly increased in LAQ824-treated HT29 xenografts, and this ratio was inversely correlated with changes in tumor volume. Statistically significant decreases in intracellular pH, beta-nucleoside triphosphate (beta-NTP)/TotP, and beta NTP/inorganic phosphate (Pi) and an increase in Pi/TotP were also seen in LAQ824 treated tumors. Tumor extracts showed many significant metabolic changes after LAQ824 treatment, in parallel with increased histone acetylation and decreased microvessel density. Treatment with LAQ824 resulted in altered phospholipid metabolism and compromised tumor bioenergetics. The phosphocholine and phosphomonoester increases may have the potential to act as pharmacodynamic markers for noninvasively monitoring tumor response after treatment with LAQ824 or other HDAC inhibitors. PMID- 18392141 TI - An in vivo mouse model for human prostate cancer metastasis. AB - We developed a sensitive real-time polymerase chain reaction (QPCR) assay that allows us to track early lodging/homing events in vivo. We used this technology to develop a metastasis assay of human prostate cancer (PCa) growth in severe combined immunodeficient mice. For this purpose, marked human PCa cell lines were implanted subcutaneously or in the prostate (orthotopically) of severe combined immunodeficient mice as models of primary tumors. Mice were then sacrificed at various time points, and distant tissues were investigated for the presence of metastatic cells. At 3 weeks, a number of tissues were recovered and evaluated by QPCR for the presence of metastatic cells. The data demonstrate that several PCa cell lines are able to spread from the primary lesion and take up residence in distant sites. If the primary tumors were resected at 3 weeks, in several cases, metastatic lesions were identified over the course of 9 months. We propose that this new model may be particularly useful in exploring the molecular events in early metastasis, identifying the metastatic niche, and studying issues pertaining to dormancy. PMID- 18392142 TI - Increased mitochondrial DNA damage and down-regulation of DNA repair enzymes in aged rodent retinal pigment epithelium and choroid. AB - PURPOSE: In the central nervous system (CNS), increased mitochondrial DNA (mtDNA) damage is associated with aging and may underlie, contribute to, or increase the susceptibility to neurodegenerative diseases. Because of the focus on the retinal pigment epithelium (RPE) and choroid as tissue relevant to age-related macular degeneration (AMD), we examined young and aged RPE and choroid, harvested from rodent eyes, for DNA damage and for changes in selected DNA repair enzymes. METHODS: Immunohistochemical labeling and quantitative ELISA for the oxidative DNA damage marker, 8-hydroxy-2'-deoxy-guanosine (8-OHdG), were measured in young and aged rodent RPE and choroid. mtDNA and nuclear DNA (nDNA) damage was determined by quantitative polymerase chain reaction (PCR) by comparing the relative amplification of small and large DNA fragments. Expression of several DNA repair enzymes was measured using real-time quantitative reverse transcription -PCR (qRT-PCR) and immunoblot. RESULTS: Immunohistochemical labeling for 8-OHdG increased in aged rodent RPE and choroid. Quantitative ELISA confirmed increased levels of 8-OHdG. Measurements of nDNA and mtDNA lesions indicated that DNA damage is primarily in mtDNA in aged RPE and choroid. Using qRT-PCR, we found that gene expression of DNA repair enzymes, 8-oxoguanine-DNA glycosylase 1 (OGG1), mutY homolog (MYH), and thymine DNA glycosylase were decreased in an age-dependent pattern in RPE and choroid. However, endonuclease III homolog 1 was not significantly changed in aged RPE and choroid. Using immunoblots, we found that protein levels of OGG1 and MYH were decreased in aged RPE and choroid. CONCLUSIONS: Our results show that there is increased mtDNA damage in aged RPE and choroid, which is likely due to decreased DNA repair capability. mtDNA damage in the RPE and choroid may be a susceptibility factor that underlies the development of AMD. PMID- 18392143 TI - Interactions between SNP alleles at multiple loci contribute to skin color differences between caucasoid and mongoloid subjects. AB - This study aimed to identify single nucleotide polymorphism (SNP) alleles at multiple loci associated with racial differences in skin color using SNP genotyping. A total of 122 Caucasians in Toledo, Ohio and 100 Mongoloids in Japan were genotyped for 20 SNPs in 7 candidate genes, encoding the Agouti signaling protein (ASIP), tyrosinase-related protein 1 (TYRP1), tyrosinase (TYR), melanocortin 1 receptor (MC1R), oculocutaneous albinism II (OCA2), microphthalmia associated transcription factor (MITF), and myosin VA (MYO5A). Data were used to analyze associations between the 20 SNP alleles using linkage disequilibrium (LD). Combinations of SNP alleles were jointly tested under LD for associations with racial groups by performing a chi(2) test for independence. Results showed that SNP alleles at multiple loci can be considered the haplotype that contributes to significant differences between the two population groups and suggest a high probability of LD. Confirmation of these findings requires further study with other ethnic groups to analyze the associations between SNP alleles at multiple loci and skin color variation among races. PMID- 18392144 TI - Expression and function of micro-RNAs in immune cells during normal or disease state. AB - Micro-RNAs (miRNAs) are 19-24 nucleotide long non-coding RNAs that posttranscriptionally modulate gene expression. They are found in almost all species: viruses, plants, nematodes, fly, fish, mouse, human, and are implicated in a wide array of cellular and developmental processes. Microarray-based miRNA profiling brought to the discovery of miRNAs specific to different hematopoietic lineages. Furthermore, the functional assays performed in tissue cultures to discover miRNAs involved in immune responses in combination with the reports of miRNA-transgenic or miRNA -knockout mouse models has helped elucidating the miRNA roles in the development and function of immune system. Abnormal patterns of hematopoietic-specific miRNAs have been found in different types of cancer and miRNA based gene therapy is being considered as a potential technology of choice in immunological disorders and cancer. The purpose of this review is to discuss recent findings related with the expression and function of miRNAs in hematopoietic lineages. PMID- 18392145 TI - A 12 week, open label, phase I/IIa study using apatone for the treatment of prostate cancer patients who have failed standard therapy. AB - PURPOSE: To evaluate the safety and efficacy of oral Apatone (Vitamin C and Vitamin K3) administration in the treatment of prostate cancer in patients who failed standard therapy. MATERIALS AND METHODS: Seventeen patients with 2 successive rises in PSA after failure of standard local therapy were treated with (5,000 mg of VC and 50 mg of VK3 each day) for a period of 12 weeks. Prostate Specific Antigen (PSA) levels, PSA velocity (PSAV) and PSA doubling times (PSADT) were calculated before and during treatment at 6 week intervals. Following the initial 12 week trial, 15 of 17 patients opted to continue treatment for an additional period ranging from 6 to 24 months. PSA values were followed for these patients. RESULTS: At the conclusion of the 12 week treatment period, PSAV decreased and PSADT increased in 13 of 17 patients (p < or = 0.05). There were no dose-limiting adverse effects. Of the 15 patients who continued on Apatone after 12 weeks, only 1 death occurred after 14 months of treatment. CONCLUSION: Apatone showed promise in delaying biochemical progression in this group of end stage prostate cancer patients. PMID- 18392146 TI - Principal investigator views of the IRB system. AB - We undertook a qualitative e-mail survey of federally-funded principal investigators of their views of the US human subjects protection system, intended to identify the range of investigator attitudes. This was an exploratory study with a 14% response rate. Twenty-eight principal investigators responded; their comments were analyzed to show underlying themes, which are here presented along with supporting quotations.There was consensus that it is important to protect human subjects from research abuse, but disagreement over how well the IRB system is functioning. Some researchers felt that the system is effective and serves its purpose well. Of those who support the system, some endorse its methods, purpose, and daily functioning, as they experience it, without reservation. Others, while expressing some frustration, feel that the purpose is important and their local IRB does its best to make a difficult system work well.Those investigators who were more harshly critical commented on multiple flaws in the system, including (1) consent forms that are inappropriate and incomprehensible, (2) an emphasis on minutiae, and (3) concern with protecting the institution more than research subjects. Respondents told us that the IRB system is a particular burden for research in neurology, emergency medical conditions, repositories, and social sciences in general; a more comprehensive study might identify other problematic areas. Significant concern was expressed about the cost, inefficiency, and irrationality of IRB review. The IRB system works well for some researchers, but our results indicate that other investigators feel the costs outweigh the benefits. PMID- 18392147 TI - Biosynthetic potentials of metabolites and their hierarchical organization. AB - A major challenge in systems biology is to understand how complex and highly connected metabolic networks are organized. The structure of these networks is investigated here by identifying sets of metabolites that have a similar biosynthetic potential. We measure the biosynthetic potential of a particular compound by determining all metabolites than can be produced from it and, following a terminology introduced previously, call this set the scope of the compound. To identify groups of compounds with similar scopes, we apply a hierarchical clustering method. We find that compounds within the same cluster often display similar chemical structures and appear in the same metabolic pathway. For each cluster we define a consensus scope by determining a set of metabolites that is most similar to all scopes within the cluster. This allows for a generalization from scopes of single compounds to scopes of a chemical family. We observe that most of the resulting consensus scopes overlap or are fully contained in others, revealing a hierarchical ordering of metabolites according to their biosynthetic potential. Our investigations show that this hierarchy is not only determined by the chemical complexity of the metabolites, but also strongly by their biological function. As a general tendency, metabolites which are necessary for essential cellular processes exhibit a larger biosynthetic potential than those involved in secondary metabolism. A central result is that chemically very similar substances with different biological functions may differ significantly in their biosynthetic potentials. Our studies provide an important step towards understanding fundamental design principles of metabolic networks determined by the structural and functional complexity of metabolites. PMID- 18392148 TI - Statistical resolution of ambiguous HLA typing data. AB - High-resolution HLA typing plays a central role in many areas of immunology, such as in identifying immunogenetic risk factors for disease, in studying how the genomes of pathogens evolve in response to immune selection pressures, and also in vaccine design, where identification of HLA-restricted epitopes may be used to guide the selection of vaccine immunogens. Perhaps one of the most immediate applications is in direct medical decisions concerning the matching of stem cell transplant donors to unrelated recipients. However, high-resolution HLA typing is frequently unavailable due to its high cost or the inability to re-type historical data. In this paper, we introduce and evaluate a method for statistical, in silico refinement of ambiguous and/or low-resolution HLA data. Our method, which requires an independent, high-resolution training data set drawn from the same population as the data to be refined, uses linkage disequilibrium in HLA haplotypes as well as four-digit allele frequency data to probabilistically refine HLA typings. Central to our approach is the use of haplotype inference. We introduce new methodology to this area, improving upon the Expectation-Maximization (EM)-based approaches currently used within the HLA community. Our improvements are achieved by using a parsimonious parameterization for haplotype distributions and by smoothing the maximum likelihood (ML) solution. These improvements make it possible to scale the refinement to a larger number of alleles and loci in a more computationally efficient and stable manner. We also show how to augment our method in order to incorporate ethnicity information (as HLA allele distributions vary widely according to race/ethnicity as well as geographic area), and demonstrate the potential utility of this experimentally. A tool based on our approach is freely available for research purposes at http://microsoft.com/science. PMID- 18392151 TI - Emergent perspectives on ADHD: sharing knowledge and extending care. PMID- 18392149 TI - When the genome plays dice: circumvention of the spindle assembly checkpoint and near-random chromosome segregation in multipolar cancer cell mitoses. AB - BACKGROUND: Normal cell division is coordinated by a bipolar mitotic spindle, ensuring symmetrical segregation of chromosomes. Cancer cells, however, occasionally divide into three or more directions. Such multipolar mitoses have been proposed to generate genetic diversity and thereby contribute to clonal evolution. However, this notion has been little validated experimentally. PRINCIPAL FINDINGS: Chromosome segregation and DNA content in daughter cells from multipolar mitoses were assessed by multiphoton cross sectioning and fluorescence in situ hybridization in cancer cells and non-neoplastic transformed cells. The DNA distribution resulting from multipolar cell division was found to be highly variable, with frequent nullisomies in the daughter cells. Time-lapse imaging of H2B/GFP-labelled multipolar mitoses revealed that the time from the initiation of metaphase to the beginning of anaphase was prolonged and that the metaphase plates often switched polarity several times before metaphase-anaphase transition. The multipolar metaphase-anaphase transition was accompanied by a normal reduction of cellular cyclin B levels, but typically occurred before completion of the normal separase activity cycle. Centromeric AURKB and MAD2 foci were observed frequently to remain on the centromeres of multipolar ana-telophase chromosomes, indicating that multipolar mitoses were able to circumvent the spindle assembly checkpoint with some sister chromatids remaining unseparated after anaphase. Accordingly, scoring the distribution of individual chromosomes in multipolar daughter nuclei revealed a high frequency of nondisjunction events, resulting in a near-binomial allotment of sister chromatids to the daughter cells. CONCLUSION: The capability of multipolar mitoses to circumvent the spindle assembly checkpoint system typically results in a near-random distribution of chromosomes to daughter cells. Spindle multipolarity could thus be a highly efficient generator of genetically diverse minority clones in transformed cell populations. PMID- 18392150 TI - Metaproteomics provides functional insight into activated sludge wastewater treatment. AB - BACKGROUND: Through identification of highly expressed proteins from a mixed culture activated sludge system this study provides functional evidence of microbial transformations important for enhanced biological phosphorus removal (EBPR). METHODOLOGY/PRINCIPAL FINDINGS: A laboratory-scale sequencing batch reactor was successfully operated for different levels of EBPR, removing around 25, 40 and 55 mg/l P. The microbial communities were dominated by the uncultured polyphosphate-accumulating organism "Candidatus Accumulibacter phosphatis". When EBPR failed, the sludge was dominated by tetrad-forming alpha-Proteobacteria. Representative and reproducible 2D gel protein separations were obtained for all sludge samples. 638 protein spots were matched across gels generated from the phosphate removing sludges. 111 of these were excised and 46 proteins were identified using recently available sludge metagenomic sequences. Many of these closely match proteins from "Candidatus Accumulibacter phosphatis" and could be directly linked to the EBPR process. They included enzymes involved in energy generation, polyhydroxyalkanoate synthesis, glycolysis, gluconeogenesis, glycogen synthesis, glyoxylate/TCA cycle, fatty acid beta oxidation, fatty acid synthesis and phosphate transport. Several proteins involved in cellular stress response were detected. CONCLUSIONS/SIGNIFICANCE: Importantly, this study provides direct evidence linking the metabolic activities of "Accumulibacter" to the chemical transformations observed in EBPR. Finally, the results are discussed in relation to current EBPR metabolic models. PMID- 18392152 TI - Knowledge exchange for Attention Deficit Hyperactivity Disorder Research: an integrated evidence and knowledge exchange framework leading to more effective research dissemination practices. AB - INTRODUCTION: Knowledge Exchange refers to activities that help to create and support the conditions and culture that lead to the most effective access, implementation, utilization, and evaluation of the most credible evidence for improved mental health outcomes for children and youth in Ontario. Although knowledge exchange and associated concepts such as knowledge transfer and translation are increasingly well developed in other aspects of health and healthcare, it is underdeveloped in mental health generally. This paper introduces some of the basic concepts of knowledge exchange and calls for more development of knowledge exchange in the area of Attention Deficit Hyperactivity Disorder Research. METHODS: This is a discussion paper that presents a general overview of the Centre's approach to knowledge exchange. It links the discussion to related concepts and to the need to overcome the research to practice gap. The Integrated Evidence and Knowledge Exchange Framework of the Provincial Centre of Excellence for Child and Youth Mental Health is introduced. Areas of active development in knowledge exchange are categorized into three objectives: context, content, and capacity. RESULTS: The use of an Integrated Evidence and Knowledge Exchange Framework for the Centre's Grants and Awards program activities and evaluation has begun to explicitly and transparently link the evidence on effective knowledge exchange with the evidence on effective treatment for children and youth with mental health difficulties including ADHD. This framework is expected to produce greater transparency as well as improved attainment of outputs, outcomes, and impacts of these grants and awards in child and youth mental health. CONCLUSIONS: Knowledge exchange activities may reduce the confusion for parents & care-givers, practitioners, researchers, and administrators, seeking the most credible data, information and knowledge about the most effective treatments for ADHD. An active process that seeks to improve knowledge exchange for ADHD is needed. PMID- 18392153 TI - Can PLAY diminish ADHD and facilitate the construction of the social brain? AB - The diagnosis of attention deficit hyperactivity disorders (ADHD) has been increasing at an alarming rate, paralleled by the prescription of highly effective psychostimulants whose developmental effects on growing brains remain inadequately characterized. One reason for the increasing incidence of ADHD may be the diminishing availability of opportunities for pre-school children to engage in natural self-generated social play. Pre-clinical work indicates that play can facilitate behavioral inhibition in growing animals, while psychostimulants reduce playfulness. The idea that intensive social play interventions, throughout early childhood, may alleviate ADHD symptoms remains to be evaluated. As an alternative to the use of play-reducing psychostimulants, society could establish play "sanctuaries" for at-risk children in order to facilitate frontal lobe maturation and the healthy development of pro-social minds. PMID- 18392154 TI - Attention-Deficit/Hyperactivity Disorder and the challenges of close friendship. AB - INTRODUCTION: Much has been learned about the social rejection of children with Attention-Deficit Hyperactivity Disorder (ADHD) by their schoolmates. Although these group processes are clearly important, recent advances in theory and research have revealed the importance of close friendship. METHOD: In this paper, we review the current knowledge about the close friendships of children with ADHD RESULTS: Although children with ADHD tend to be excluded from close friendship, the data on the features of the friendships they do have are too limited and too flawed to permit conclusions about patterns of inter-action between children with ADHD and their friends. Few data are actually available to indicate why children with ADHD have difficulty keeping the friends they do have. CONCLUSION: We conclude by briefly discussing some implications for pharmacological treatments and peer-relations interventions. PMID- 18392155 TI - Attention-deficit/hyperactivity disorder and oppositional/conduct problems: links to parent-child interactions. AB - INTRODUCTION: We outline a transactional model of parent-child interactions which proposes that characteristics of attention-deficit/hyperactivity disorder (ADHD) appear early in a child's life and present challenges to the parent's ability to maintain consistent and responsive parenting. In turn, the presence of harsh or inconsistent parenting is hypothesized to contribute to the development of child oppositional behaviour and conduct problems. METHOD AND RESULTS: We briefly discuss existing cross-sectional evidence in support of this model and then highlight five longitudinal studies that provide stronger support, showing that difficulties in parenting predict the development of oppositional and conduct problems in children with ADHD, over and above pre-existing levels of child problems and other predictors. CONCLUSION: We conclude with a discussion of the clinical implications of this model and evidence, emphasizing the need for early interventions in ADHD focused on parenting. PMID- 18392156 TI - French version of the strengths and weaknesses of ADHD symptoms and normal behaviors (SWAN-F) questionnaire. AB - OBJECTIVE: To evaluate internal and external consistency of a French adaptation of the SWAN (a 7-point rating strength-based scale, from far below to far above average) and its accuracy as a diagnostic test among children with Attention Deficit/Hyperactivity Disorder (ADHD). METHOD: Parents of 88 children referred for ADHD were interviewed using the SWAN-F, a structured interview (DISC-4.0) and the Conners' Rating Scale. Internal consistency and divergent and convergent validity of the SWAN-F were examined using the DISC-4.0 and Conners' Rating Scales as reference standards for four dimensions: Inattention, Hyperactivity/Impulsivity, ADHD, Oppositional Defiant Disorder. RESULTS: The internal consistency of SWAN-F was within acceptable ranges for all dimensions (Cronbach's alpha greater than 0.80). Scores of the SWAN-F subscales were strongly associated with the DISC-4.0 diagnostic assignments and Conners' Rating Scales, following logical patterns of correspondence between diagnoses. Its accuracy as a diagnostic test was comparable to Conners' Rating Scale, with a lower rate of false positives. CONCLUSIONS: The information gathered with the SWAN-F is compatible with that obtained using the DISC-4.0 and Conners' Rating Scale. Strength-based rating scales have the potential to evaluate the normal distribution of behaviors and to provide reliable cut-off defining abnormal behavior. PMID- 18392158 TI - Clinical case rounds in child and adolescent psychiatry selective mutism. PMID- 18392157 TI - Comparison of the predictive validity of hyperkinetic disorder and attention deficit hyperactivity disorder. AB - INTRODUCTION: We compared the predictive validity of attention deficit hyperactivity disorder (ADHD; Diagnostic and Statistical Manual - IV Edition) and hyperkinetic disorder (HKD; International Classification of Diseases - 10th Edition) while controlling for the presence of comorbid psychopathology. METHOD: ADHD and HKD criteria were used to classify 804 clinic-referred children ages 6 to 16 years into one of four non-overlapping groups: HKD, ADHD combined subtype (ADHD-C), ADHD hyper-active-impulsive subtype (ADHD-HI), ADHD inattentive subtype (ADHD-IA). Groups were compared with each other and with normal controls (67) while controlling for age and intelligence on a range of criteria both before and after excluding cases with comorbidity. RESULTS: Of the 804 clinic participants, 72 (8.9 %) met criteria for ICD-10 HKD, 353 (43.9 %) for ADHD-C, 142 (17.7 %) for ADHD-HI and 237 (29.5 %) for ADHD-IA. There were no differences among the four clinic groups in rate of comorbidity, neuro-developmental or psychosocial risk indices, inter-parental or parent-child discord, family history of ADHD, working memory, and academic or intelligence test scores, but all clinic groups differed from normal controls. By contrast, total number of symptoms, teacher-rated impairment and inhibitory control deficit were greatest in HKD and least in ADHD C, ADHD-HI, ADHD-IA in that order. Results of the comparisons were essentially unchanged after excluding cases (75%) with a comorbid condition. CONCLUSIONS: HKD, ADHD-C, ADHD-HI and ADHD-IA had approximately equivalent predictive validity even when comorbidity was taken into account. PMID- 18392159 TI - Clinical case rounds in child and adolescent psychiatry: commentary on selective mutism. PMID- 18392160 TI - Evidence-based family psychoeducational interventions for children and adolescents with psychotic disorders. AB - INTRODUCTION: Family psychoeducational interventions have consistently been found to impact families positively and reduce relapse rates in individuals with psychotic disorders. Research finds that, for adults, family psychoeducational interventions are effective in preventing relapse and improving social and occupational functioning. Psychotic disorders are increasingly recognized as having early onset, yet limited psychoeducational evidence-based intervention services are available and no intervention has centered exclusively on youth with a psychotic disorders and their families. METHOD: This article reviews the evidence-based literature on family psychoeducational interventions for persons with a psychotic disorder, with a specific focus on the gaps, strengths, and limitations of family psychoeducational treatment for children and adolescents. This article incorporates current research in the proposed development of a family psychoeducational intervention exclusively for adolescents with a psychotic disorder and their parents. RESULTS: A conceptual psychoeducational multiple family group intervention (PMFG) for adolescents with a psychotic disorder is presented. CONCLUSION: The impact of these disorders affects not only the diagnosed adolescents and their families, but places a significant burden on the health care system and society. This article adapts an evidence-based intervention to improve prognosis, social and peer functioning, and reduce relapse in children and adolescents throughout their life cycle. PMID- 18392161 TI - Evaluation of a community-based program for young boys at-risk of antisocial behaviour: results and issues. AB - OBJECTIVE: We assess the impact of a community-based intervention program for boys 6-11 years old at-risk of antisocial behaviour, and compare changes in behaviour and competence pre-post for intervention and wait-list comparison group. METHOD: Interested parents called for enrolment. Inclusion required police contact and/or clinical scores (T>69) on Child Behaviour Checklist (CBCL) or Teacher Report Form (TRF), no developmental delay and English speaking. The program included two core 12-week groups (children's, parents') and optional additional services. Twelve sessions (February 2002-December 2005) provide pre post intervention data, boys waiting at least 6 months formed a comparison group (starting April 2005). Outcomes included CBCL and TRF behaviour scales (rule breaking, aggression, conduct, total problems) and competence. Repeated measures analysis of variance was done. RESULTS: Pre-post outcome comparisons indicated improvements among all boys, with significant differences favouring intervention boys on CBCL behaviour scales, but not TRF outcomes. Effect sizes were small to medium. Persisting high post-behaviour levels, unmeasured variation in additional services, and other design and sampling issues are noted. CONCLUSIONS: More rigorously designed program evaluation is required. PMID- 18392162 TI - Where do young children in specialty care come from?: a preliminary investigation of the role of primary care physicians. AB - INTRODUCTION: Young children with mental health and developmental problems may not receive appropriate or timely interventions. Primary care physicians are well positioned to play an important role in the early identification and referral of such children. The objective of this pilot study was to explore the role primary care physicians played for a group of such children. METHODS: A single mailing of 1196 self-report questionnaires were sent to parents/caregivers of children under six years of age at four specialty centers in Ontario and Alberta. Key items on the survey included the role of primary care physicians (family physicians and community paediatricians). RESULTS: Twenty percent of parents/caregivers returned questionnaires. All children saw either a family physician or a community paediatrician, while 65% saw both. Families were more likely to have come to the specialty centre via a referral from a community paediatrician than a family physician. Ten percent reportedly received no referrals from a primary care physician, while 21% did not receive a referral to a specialty centre from these providers. CONCLUSIONS: The majority of children received at least one referral from a primary care physician. Further inquiry is required to determine the timeliness and appropriateness of these referrals. PMID- 18392163 TI - Separation, autism, and residential treatment. PMID- 18392164 TI - Commentary on Separation, autism, and residential treatment: tapping the strengths of the ASD parent. PMID- 18392165 TI - New developments in child and adolescent eating disorders. PMID- 18392166 TI - A review of psychotherapeutic interventions for children and adolescents with eating disorders. AB - OBJECTIVE: Psychotherapeutic interventions for child and adolescent eating disorders have recently received increasing attention in the research literature. This article attempts to summarize these studies. METHOD: The current literature was reviewed using the PubMed and Embase databases under the search terms eating disorders, child, adolescent, and psychotherapy. Here we will present a practical overview of the current evidence for psychotherapeutic interventions in this clinical population. RESULTS: There have been some very promising findings with regards to specific types of therapy for anorexia nervosa (AN), bulimia nervosa (BN) and binge eating disorder (BED). CONCLUSIONS: The best evidence available suggests that family-therapy models are most effective for treating adolescents with AN, and that CBT models are most effective for adolescent BN, although family-based treatment may also be effective for adolescents with BN. Too few studies have been done on BED in adolescents to draw any conclusions; however CBT, IPT and DBT are all theoretically promising. PMID- 18392167 TI - Self-silencing in a clinical sample of female adolescents with eating disorders. AB - OBJECTIVE: The present study was designed to assess the relationship between self silencing behaviours and eating disorder symptoms in a female adolescent population with eating disorders. METHOD: One hundred and forty-nine adolescent girls between the ages of 13 and 18 completed a comprehensive assessment at a tertiary care children's hospital. Each participant completed the Eating Disorder Inventory-2 (EDI-2; Garner, 1991), the Multidimensional Anxiety Scale for Children (MASC; March et al, 1997), and an adapted version of the Silencing the Self Scale for adolescents (STSS; Sippola & Bukowski, 1996). RESULTS: Self silencing behaviours correlated strongly with eating disorder symptomatology. Social anxiety was found to predict body dissatisfaction, while externalized self perception was found to contribute uniquely to body dissatisfaction and drive for thinness, two risk factors closely associated with eating disorders. CONCLUSION: These results suggest the importance of including relational and emotional development in comprehensive models of disordered eating. PMID- 18392168 TI - Evaluation of meal support training for parents and caregivers using a video and a manual. AB - OBJECTIVE: The purpose of the study was to evaluate the helpfulness of the contents of a video and manual for training parents and caregivers in providing meal support for eating disordered youth. METHOD: A self-report questionnaire consisting of closed and open-ended questions was given parents or caregivers of 52 new consecutive referrals to the specialized eating disorder program at British Columbia Children's Hospital. RESULTS: The return rate was 77%. Sixty five percent of the families were dealing with a youth diagnosed with anorexia nervosa, 33%, eating disorder not otherwise specified, and 2%, bulimia nervosa. The meal support training resources were deemed informative by the parents who reviewed the material. Parents most valued hearing about the youth's experience of the eating disorder and their feelings around food. CONCLUSION: As a minimum initial intervention, the combination of the manual and DVD/video was found to be convenient to use and was well received by families with eating disordered youth. These resources could be used by clinician treating an eating disordered youth and his or her family in order to start addressing the food issues. PMID- 18392169 TI - Challenges associated with controlled psychopharmacological research trials in adolescents with eating disorders. AB - INTRODUCTION: Eating disordered populations present many unique challenges both to clinicians and researchers. Adolescents with eating disorders can be difficult to treat, and the challenges associated with research in this area can be significant. OBJECTIVES: This paper was written with three main objectives in mind: to comment on some of the barriers impeding mental health research in general, to highlight challenges faced in the design and implementation of ED specific studies, and to integrate personal insight into some of the many challenges that we have encountered during our experience with a randomized clinical trial examining the efficacy of olanzapine for the adjunctive treatment of youth with Anorexia Nervosa. DISCUSSION: It is hoped that providing information in this context will allow researchers greater insight into some of the many challenges that accompany study of this cohort. PMID- 18392170 TI - A review of medication use for children and adolescents with eating disorders. AB - OBJECTIVE: This paper aims to review the research literature on the use of medication for eating disorders in children and adolescents. METHOD: The literature was reviewed on the pharmacotherapy of anorexia nervosa (AN), bulimia nervosa (BN) and eating disorder not otherwise specified (EDNOS). The PubMed database was searched for all articles on medication use in the child and adolescent population using the terms medication, antipsychotic, antidepressant, child, adolescent, eating disorders, anorexia nervosa and bulimia nervosa. RESULTS: Very little literature exists on the use of medication for the treatment of eating disorders in children and adolescents. There is one retrospective study on the use of SSRIs and some case reports on atypical antipsychotics for children and adolescents with AN, and one small open trial on SSRIs for adolescent BN. CONCLUSIONS: Evidence-based pharmacological treatment for children and adolescents with eating disorders is not yet possible due to the limited number of studies available. It appears that olanzapine and other atypical antipsychotics may prove to be promising for AN at low body weights. It remains uncertain whether SSRIs are helpful in preventing relapse in AN. For children and adolescents with BN, the first line pharmacological option is fluoxetine given the large evidence base of this drug with the adult population and a small open trial of adolescents with BN. PMID- 18392171 TI - Compulsive hoarding associated with abortion. PMID- 18392172 TI - Commentary on Compulsive hoarding associated with abortion. PMID- 18392173 TI - Lamotrigine in adolescent mood disorders: a retrospective chart review. AB - INTRODUCTION: Treatment nonresponse in adolescent mood disorders is a major public health problem, as mood disorders in youth are associated with significant mortality by suicide, protracted course of illness, and recurrence into adulthood. Three studies with small sample sizes exist for lamotrigine in youth mood disorders. However, the risk of serious rash has limited its use in youth mood disorders. OBJECTIVE: The aims of this study are to evaluate the preliminary effectiveness and safety of lamotrigine in adolescent mood disorders. METHODS: Medical charts were retrospectively reviewed at three clinical sites for 42 adolescents treated with lamotrigine for a mood disorder. The Clinical Global Impression (CGI) Severity and Improvement scores were obtained at baseline and last visit. Treatment-emergent adverse effects were also obtained. RESULTS: Improvement was seen in 22 subjects (52%). The mean daily lamotrigine dose was 114.8mg (SD 75.6), and the average duration of lamotrigine treatment was 29.1+/ 31.8 weeks. The mean CGI-S score decreased from 4.9+/-1.0 at baseline to 3.5+/ 1.4 at endpoint (z=3.204, p<0.002). Four subjects (10%) developed benign rash. CONCLUSIONS: This study provides preliminary data that lamotrigine may be effective in adolescents with mood disorders. However, this study revealed that lamotrigine might be associated with a significant risk of benign rash. PMID- 18392174 TI - [Risk factors in families of children with disorder attention deficit/hyperactivity: Quebec data]. AB - INTRODUCTION: This paper focuses on Attention-Deficit/Hyperactivity Disorder (ADHD) and links with other disorders in the child and his/her parents. Adversity factors are presented around the family life and their impact on ADHD. Families who have a child with ADHD are compared to families who do not. METHOD: The parents in 82 families filled in the QFR-ADHD questionnaire: 24 children without ADHD (control group) and 58 children with ADHD (ADHD group). The children were between 5 and 21 years of age (average age: 10 years) with an average education of 1 to 14 years (average: 4th year of elementary school). The subjects were distributed as follows: 9 boys and 15 girls (control group), and 48 boys and 10 girls (TDA/H group). RESULTS: Oppositional disorder (OD), conduct disorders (CD) and learning disabilities (LD) were significantly more frequent in the ADHD group than in the control group. We found that the mothers of children with ADHD take more selective serotonin reuptake inhibitors than the mothers of children in the control group. This implies that the former experience depressive symptoms. Fathers of children with ADHD had more learning disabilities than the fathers in the control group. CONCLUSION: This research is in line with work on the etiology of ADHD. Investigation of the causes for ADHD is complex, as the disorder has both physical and psychological aspects. PMID- 18392175 TI - Priority-setting for children's mental health: clinical usefulness and validity of the priority criteria score. AB - OBJECTIVE: The 17-item PCS was designed for priority-setting and queue management of children and adolescents referred for mental health services. Here we assess aspects of the validity of the Children's Mental Health (CMH) Priority Criteria Score (PCS), developed by the Western Canada Waiting List Project (WCWL). The PCS was evaluated across clinical settings of increasing acuity and in terms of its relationship to two variables reflecting criteria-related validity and actual wait times. METHOD: Intake workers completed PCS forms for 497 referrals enrolled for treatment in three clinical areas over approximately two fiscal years. The completion time of the PCS form was estimated in relation to the total referral and screening process. Intake workers completed the PCS items and did not use the total score at the time of intake and form completion to triage or place clients; hence, the PCS was independent of enrollment and placement within the continuum of care. Furthermore, clinicians in the receiving programs had to accept the triage decisions for the PCS to be used in the study analysis. RESULTS: The PCS score was meaningfully related to the measures of criteria-related validity (e.g., clinician perceived urgency, clinician perceived maximum acceptable waiting times) and triage to clinical settings of increasing acuity. There was a significant mean difference in the PCS for those accepted to community, day, or inpatient settings. CONCLUSIONS: The PCS appears to be a useful, efficient measure of clinical urgency adequate for use in priority-setting for children waiting for mental health services. PMID- 18392176 TI - Corticosteroid-related psychiatric complications in the treatment of Hodgkin's lymphoma in an adolescent. PMID- 18392177 TI - Commentary on Corticosteroid-related psychiatric complications in the treatment of Hodgkin's lymphoma in an adolescent. PMID- 18392179 TI - Commentary on Treatment Resistant Psychosis in an adolescent with scoliosis and a history of early feeding difficulties. PMID- 18392181 TI - What we know about ADHD and driving risk: a literature review, meta-analysis and critique. AB - INTRODUCTION: This article examines the literature on ADHD and unintentional driving injury. This literature has emerged over the last decade as part of the burgeoning epidemic of road traffic death and injury which is the number one cause of death in young adults in North America. METHODS: The available literature on observational outcome studies and experimental pharmacological interventions is critically reviewed. A meta-analysis of behavioral outcomes and a review of effect size of pharmacological studies are presented. RESULTS: Current data support the utility of stimulant medication in improving driving performance in younger ADHD drivers. A conceptual model of risk factors in young ADHD drivers is offered. CONCLUSION: The current state of screening instruments for identifying high risk subjects within this clinical group is summarized along with a final section on emerging trends and future prospects for intervention. PMID- 18392182 TI - A methodology for conducting retrospective chart review research in child and adolescent psychiatry. AB - INTRODUCTION: Retrospective research has become largely undervalued and underutilized in child and adolescent psychiatry with the increasing singular focus on randomized control trials, despite the wealth of clinically relevant data available in historical medical records. In this paper a systematic and scientific approach to chart review research methodology for psychiatry is described. METHOD: Informed by available literature, a methodological stepwise approach for retrospective chart review was developed. RESULTS: A nine step method aimed at maximizing benefits and minimizing limitations is discussed. CONCLUSIONS: Retrospective chart review is an important methodology with distinct advantages and has the potential to provide psychiatry with valuable research opportunities. This method of study should not be lost in the field of psychiatry. PMID- 18392183 TI - Treatment refusal in adolescents with severe chronic illness and borderline personality disorder. AB - INTRODUCTION: The aim of this study is to address the complex psychopathologic factors involved in treatment refusal observed in adolescents suffering from a severe chronic illness. METHOD: We report on five chronically ill adolescents (2 diabetes mellitus, 1 maple syrup urine disease, 1 bird fancier's lung, 1 HIV infection) who were consecutively admitted to an inpatient psychiatric service as a result of a life-threatening refusal to comply with outpatient management of their medical illness. Case material is analyzed and discussed in the context of a review of the literature. RESULTS: Each subject was further characterized by: (1) the diagnosis of Borderline Personality Disorder; (2) severe family dysfunction (e.g. abuse, neglect and abandonment); and (3) frequent and prolonged pediatric hospitalizations. During their psychiatric hospitalization, the patients' initial opposition to treatment shifted to mentalization and psychological questioning. From a psychodynamic perspective (attachment theory), we hypothesized that this behavior represented an attack on parental figures as embodied in the suicidal comportment. CONCLUSION: Case material is presented to underline the possibility of co-occurring Borderline Personality Disorder when treating youths suffering from chronic illness and refusing treatment. Awareness of Borderline Personality Disorder may help pediatric staff when dealing with this refusal. PMID- 18392184 TI - Pediatric psychopharmacology update: psychostimulants and tics - past, present and future. PMID- 18392186 TI - New perspectives on intervention for conduct disorder. PMID- 18392188 TI - Is there bias in the criteria used to judge submissions to the 2005 AACAP conference? PMID- 18392187 TI - Introducing the new Journal of the Canadian Academy of Child and Adolescent Psychiatry. PMID- 18392189 TI - Response to the letter to the Editor. PMID- 18392190 TI - Conduct disorder: new directions in promoting effective parenting and strengthening parent-adolescent relationships. AB - INTRODUCTION: Although it is always preferable to prevent childhood mental health and behavioural problems, it is not always possible to do so early in their developmental trajectory. Adolescence offers another window of opportunity to intervene and reduce risk or prevent the development of late onset conditions. In this paper, we focus on adolescent Conduct Disorder, noting the special challenges of this developmental period and implications for interventions. We briefly discuss the growing evidence of the importance of parent and family targeted interventions. METHOD: Forty-eight adolescents with severe Conduct Disorder and their caregivers completed questionnaires at the beginning and end of a parent group program (Connect Parent Group). The program is guided by attachment principles and focuses on skill development in the domains of parental attunement, empathy and effective dyadic affect regulation. These skills are the building blocks of secure attachment and important components of effective parenting. RESULTS: Caregivers' reports revealed significant improvements in their perceived parenting competence and satisfaction; reduced adolescent aggressive behaviour, internalizing and externalizing problems; and reduced levels of avoidance in the caregiver-adolescent relationship. CONCLUSION: These findings provide further support for the importance of parent and family focused interventions in the treatment of Conduct Disorder in both children and adolescents. PMID- 18392191 TI - Bullying interventions: a binocular perspective. AB - INTRODUCTION: Bullying is a complex relationship problem associated with many psychosocial difficulties for children who bully, as well as those who are victimized. A recent international volume of school-based bullying programs revealed modest effectiveness, highlighting the need to refine interventions using research on developmental profiles of children who bully and those who are victimized, as well as on their relationships. METHOD: Based on developmental systemic theory, a research review was conducted on individual and relationship risk factors associated with bullying and being victimized. RESULTS: The review led to the proposal of two organizing principles for interventions: Scaffolding and Social Architecture. Scaffolding focuses on providing tailored and dynamic supports for the needs of individual children who bully or who are victimized. Social architecture requires that adults focus on the social dynamics of children's groups and create social contexts that promote positive peer interactions and dissipate contexts that foster negative interactions. CONCLUSION: Interventions for bullying require a combination of scaffolding and social architecture to provide comprehensive supports and to change the social dynamics that enable bullying. With an empirically derived, comprehensive perspective, we may move closer to reducing the burden of these relationship problems in the lives of children and youth. PMID- 18392192 TI - Changing the social contexts of peer victimization. AB - INTRODUCTION: While school-based prevention programs often target deficits in individual children's social skills in order to limit their aggression or exposure to peer victimization, there is increasing evidence that school-wide and classroom-level factors can affect the success of these programs. METHOD: We describe the WITS Primary Program which takes a community development approach for the prevention of victimization. It was designed for kindergarten to grade 3 students, and aims to create responsive communities for the prevention of peer victimization by engaging the support of parents, teachers, school counselors, older student, and emergency services personnel. RESULTS: Evidence supporting the program's feasibility and effectiveness are reported. CONCLUSION: The prevention of peer victimization and bullying may require targeted programs with demonstrated support from many adults in young children's social networks. PMID- 18392194 TI - Nurturing hidden resilience in at-risk youth in different cultures. AB - INTRODUCTION: While there has been growing interest in the concept of resilience, there has been little attention paid to the cultural and contextual factors that influence children's healthy growth and development under adversity. Using findings from the International Resilience Project, a study of over 1500 youth in 11 countries on five continents, it has been possible to show that there are both generic and culturally specific aspects to resilience. METHOD: Fourteen communities were invited to participate based on the variability in the risks children face in each setting. A minimum of 60 youth in each community were administered the Child and Youth Resilience Measure. Qualitative interviews were also conducted with a subsample of youth. RESULTS: Both homogeneity and heterogeneity in the overall sample was demonstrated, with exploratory factor analyses suggesting at least four subgroups of youth distinguished by their status as Western or non-Western, boys or girls, and the degree of social cohesion of their communities. Qualitative data explains these differences as related to seven tensions experienced by youth developmentally. CONCLUSION: This work highlights the need for greater cultural and contextual sensitivity in how resilience is understood. Implications for practice with at-risk youth include the need to understand the contextual specificity of positive development under stress. PMID- 18392195 TI - The recreation mentoring program: a community engagement initiative for children. AB - INTRODUCTION: Approximately one in five Ontario children show symptoms of significant mental health problems. These children exhibit impairments at home, at school and in the community, often with long-lasting effects. Involvement in structured community-based recreation programs may be protective for these children; however, they often encounter multiple barriers to participation (e.g., facility fees, lack of family or peer support, and reluctance to try new activities). METHOD: The Recreation Mentoring Program is a community-wide program that reduces barriers to participation while providing an important relationship with a caring, young adult mentor. Trained volunteer mentors are matched with at risk children, and meet regularly at a community recreation centre near the child's residence. The mentor's role is to: 1) stimulate participation in recreational programs, and 2) promote the child's continued participation after the mentorship ends. RESULTS: Limited program evaluation suggests that the Recreation Mentoring Program engages at-risk children in community-based recreation, that it is operationally feasible, and that it produces high levels of client satisfaction. CONCLUSION: The Recreation Mentoring Program holds promise as an effective community-based intervention for children with mental health problems. PMID- 18392196 TI - An algorithm for defining somatization in children. AB - INTRODUCTION: Defining somatization in pediatric populations presents a unique challenge, because DSM-IV somatization criteria may be inadequate for identifying a child with somatization. Two approaches exist. Child somatization has frequently been rooted in a questionnaire model, focusing on child or parent responses to assess how well a child conforms to a specific mental health profile. Others use a medical diagnosis model, designating a child with somatization as those for whom a limited number of medical measures have failed to reveal a pathological source of symptoms. METHOD: We incorporate concepts based upon a literature review from January 1994 to June 2005 of PubMed, PsycINFO, and CINAHL on classification and diagnosis of somatization in children ages 6 to 12. Our goal is to understand in depth the topic and suggest a way to better understand and classify somatization in children. RESULTS: We incorporate an integrative approach toward defining child somatization and propose an algorithm to step-by-step classify children with somatic symptoms into three distinct groups: sick, somatizers, and well. This approach includes information from self-report questionnaire, physician questionnaire, and the child's medical chart. CONCLUSION: This new algorithm suggests an approach for differentiating primary care pediatric clinic visitors into three distinct groups. Although used in clinical practice, empirical validation is necessary to further validate this algorithm. PMID- 18392193 TI - Pharmacotherapy of aggression in children and adolescents: efficacy and effect size. AB - INTRODUCTION: The treatment of pediatric aggression often involves psychotropic agents. Despite growing research on pediatric psychopharmacology, however, clinical issues regarding medication management of persistent behavioral problems remain poorly addressed. METHOD: A review of the literature from 1980 to November, 2005 yielded 45 randomized, placebo-controlled trials that addressed the treatment of aggression as either a primary or secondary outcome variable. Effect sizes (ES) (Cohen's d) were calculated for studies that met inclusion criteria. RESULTS: Overall ES for psychotropic agents in treating aggression was 0.56. Despite variability in psychiatric diagnoses, select agents showed moderate to large effects on maladaptive aggression. Most studies focused on younger children (mean age = 10.4 years), and were of short duration (7 to 70 days). Largest effects were noted with methylphenidate for co-morbid aggression in ADHD (mean ES = 0.9, combined n = 844) and risperidone for persistent behavioral disturbances in youth with conduct disorder and sub-average IQ (mean ES = 0.9, combined n = 875). CONCLUSION: A growing literature supports the use of certain medications for managing pediatric aggression. Future studies should distinguish between impulsive and predatory aggression, and examine the efficacy of agents over longer treatment periods. PMID- 18392197 TI - Using stimulant medication for children with ADHD: what do parents say? A brief report. AB - INTRODUCTION: Long-term adherence to stimulant treatment for Attention Deficit Hyperactivity Disorder (ADHD) is frequently poor. Since parents are the key decision makers regarding their child's health care, their thoughts regarding medication use are crucial to whether children take prescribed stimulants. To develop an understanding of what parents think about using stimulants to treat their children's ADHD symptoms we consulted groups of parents about their experiences. METHODS: Using qualitative methods informed by phenomenology, three focus groups of parents were asked to describe their experiences parenting a child with ADHD and using stimulant medications for treatment. Participants were 17 mothers and fathers of 14 children with ADHD, ages 7-14 years, who had received detailed diagnostic assessments and had used stimulants. Focus group dialogue was recorded and transcribed. Using established methods of data analysis, themes were identified and explored. RESULTS: Accepting that one's child has ADHD and needs treatment is a difficult and lengthy process, often accompanied by confusion and self-doubt. Parents find the choice to use stimulant treatment particularly challenging given the conflicting opinions they hear from family, friends and professionals. CONCLUSIONS: Universally, parents want to do what is best for their child. Adverse effects, concerns about stigmatization, and the child's dislike of taking pills, all contribute to parents' decisions to discontinue medication even when the child shows symptomatic benefit. PMID- 18392198 TI - Pediatric psychopharmacology update: psychostimulants and tics - past, present and future. PMID- 18392199 TI - Clinical case rounds in child and adolescent psychiatry: neurofibromatosis type 1, cognitive impairment, and attention deficit hyperactivity disorder. PMID- 18392200 TI - Commentary. PMID- 18392201 TI - Teaching children to ignore teasing: A cognitive behavioural family strategy for dealing with teasing and reactive bullying. PMID- 18392203 TI - Improving the mental health status of Canada's Aboriginal youth. PMID- 18392204 TI - Aboriginal suicidal behaviour research: from risk factors to culturally-sensitive interventions. AB - INTRODUCTION: There is a significant amount of research demonstrating that the rate of completed suicide among Aboriginal populations is much higher than in the general population. Unfortunately, there is a paucity of research evaluating the risk factors for completed suicide and suicidal behavior in this population. There is an even greater shortage of research on evidence-based interventions for suicidal behaviour. METHOD: A literature review was conducted to facilitate the development of an approach to the study of this complex problem. RESULTS: An approach to developing a research program that informs each step of the process with evidence from the previous steps was developed. The study of risk factors and interventions is described. CONCLUSIONS: Research into the risk factors and evidence-based interventions for Aboriginal suicidal behavior are required. A programmatic approach is described in detail in this paper. It is hoped this informed approach would systematically address this important public health issue that afflicts a significant proportion of the Canadian population. PMID- 18392205 TI - A community of practice approach for Aboriginal girls' sexual health education. AB - INTRODUCTION: There is a paucity of intervention programs for Aboriginal girls and many of those that exist are delivered in culturally inappropriate ways. METHODS: In this paper, we provide an overview of recent research that focused on delivering a sexual health mentorship program that enhanced the voices of Aboriginal youth and was culturally relevant and appropriate to indigenous youth. RESULTS: Our program served to enhance social connection and reinforced a sense of belonging and relational mutuality among group members. CONCLUSION: The purpose of this article is to illustrate how a mentorship program that used a community of practice approach empowered Aboriginal youth to become successful border crossers and helped to align them with the wider community. PMID- 18392206 TI - Encounters with the north: psychiatric consultation with Inuit youth. AB - INTRODUCTION: The Inuit population of Canada experiences high rates of mental health problems. Youth are particularly affected, with rates of youth suicide among the highest in the world. Psychiatric consultation is one model that has been used to address this problem. Is this the most appropriate method to assist Inuit populations with these issues? METHODS: This review addresses this question by examining literature focusing on psychiatric consultation in Canada's north, especially that which pertains to Inuit populations. RESULTS: The traditional model of medical-psychiatric consultation has significant limitations including assumptions about its validity and usefulness, its segmented approach, and its individual focus. CONCLUSION: Openness to other forms of knowledge, cultural competence, and understanding the importance of social factors are crucial in order for psychiatric consultation to be effective. PMID- 18392207 TI - Treatment resistant psychosis in an adolescent with scoliosis and a history of early feeding difficulties. PMID- 18392209 TI - [Operational criteria for diagnosis of dementia in primary care]. AB - Dementia is a chronic progressive mental disorder, which adversely affects different domains as memory, thinking, comprehension, judgment, orientation, calculation or language. People with dementia eventually lose the ability to carry out routine daily activities such as dressing, toileting, traveling and handling money. Memory impairment and dementia are commonly seen by physicians in multiple disciplines including primary care, geriatrics, neurology, and psychiatry. We discuss the pros and cons of brief cognitive tests such as the Mini-mental State Examination (MMSE) and test for executive functions to detect cognitive impairment in community patients. Diagnosis must meet standard diagnostic criteria, including tests of cognitive, global and behavioral functioning and activities of daily living. Preliminary diagnosis often takes place in primary care, usually followed by specialist assessment. In this paper we discuss operational criteria for diagnosis in primary care. An algorithm for practical physicians is proposed. PMID- 18392210 TI - [Dementia. Most frequent forms of beginning]. AB - In the present chapter, that is part of a more comprehensive work performed by the Argentine Consortium for Dementia Study - Consortium Argentino para el Estudio de la Demencia (CAED), we describe the most frequent forms of beginning for the four more prevalent types of dementia: Alzheimer's disease, dementia with Lewy bodies, vascular dementia and frontotemporal dementia). Despite this, it must be kept in mind, that frequently the clinical presentation is not typical and the diagnostic impression at the disease's beginning is controversial comparing it with the etiological diagnosis reached when the dementia is definitively installed. This issue must be considered when the initial impression is given to the patient and/or relatives. It must be clarified, in this instance, that this impression is based in statistical data of ways of presentation, but the definitive diagnosis could be different according to the dementia evolution. PMID- 18392211 TI - [Behavioural and psychological signs in dementia. Clinical features. Pharmacological and non-pharmacological treatment strategies]. AB - In first term, we define the current concepts in regard to psychosis (delirium and hallucinations) and abnormal behaviours (aggression, depression and mood changes such as mania, apathy, anxiety, agitation and desinhibition) in dementia. We also review the most used drugs in order to control these symptoms (typical and atypical antipsychotics, anti-epileptic drugs, benzodiazepines, SSRI, memantine and AcheI). As well, we take in consideration pharmacokinetic and pharmacodynamic characteristics, relationship to aging and interactions of these medications. Finally, we briefly describe the management of non-pharmacological of the most common behavioural symptoms: disruptive conducts such as exaggerated responses to minimal stimuli, catastrophic reaction, violence, anger and hostility, wandering and sundowning. As well, we discuss how to manage sleep disturbances, sexual aggression, incontinence and dressing apraxia. Management of these conditions involves, in first term, a comprehensive understanding of the whole situation and identification of underlying possible causes will make possible to evaluate results. This approach will lead to a more rationale proposal of psychotherapeutic and behavioural techniques, and milieu modifications. Finaly, we consider safety patient's in the community as well as the risk of abuse originated in a non-healthy patient-caregiver relationship. PMID- 18392212 TI - [Treatment of Alzheimer's Disease cognitive symptoms]. AB - This chapter will focus in the currently treatments for Alzheimer Disease. The meeting points in the proposed pathogenesis of the disease are the cholinergic and the cascade amyloid hypothesis based mainly in postmortem brain changes: 1- Pathological based on greater density of neuritic plaques and the characteristic presence of neurofibrillary tangles associated with neuronal loss, synaptic alterations and evidence for chronic inflammatory reactions, 2) Biochemical based on major depletion of cortical cholinergic innervation, dramatic loss in levels of biochemically determined choline acetyltransferase, accumulation of beta amyloid peptide, oxidation, glutamatergic excitotoxicity and activation of the apoptotic cascade. The currently available therapies based on the hypothesized pathophysiology of AD are: Acetylcholinesterase inhibitors (donepezil, rivastigmine and galantamine) and the NMDA receptor inhibitor Memantine. None of the "head to head" analyses done with cholinesterase inhibitors (CI) were able to demonstrate a between group effect for efficacy. However the treatment planning is based on their differences, their titration phase to reach the therapeutic doses, interactions and side effects. The non pharmacological treatment in the early and late stages of the disease, the different cognitive stimulation techniques and available prevention trials are also addressed and discussed. PMID- 18392214 TI - Web-based medical symposia. PMID- 18392213 TI - The early effects of elevated glucose on endothelial function as a target in the treatment of type 2 diabetes. AB - As predicted by the World Health Organization, the worldwide increase in the incidence of both type 1 and type 2 diabetes will result in a global increase in the impact of cardiovascular disease on human health. Both type 1 and type 2 diabetes are associated with the inability of the body to maintain appropriate levels of blood glucose. Hyperglycemia occurs when blood levels of glucose rises significantly above 5.5 mM and has been clearly linked, in particular, to microvascular complications such as retinopathy and nephropathy. Both acute and chronic hyperglycemia can result in the activation of a number of pro-oxidative and pronitrosoactive pathways. These events are activated by nuclear factor-kappa B (NF-kappabeta), protein kinase C (PKC) and nuclear enzyme poly (ADP-ribose) polymerase. Chronic hyperglycemia also results in the formation of advanced glycation end products (AGEs). Data also indicates that repetitive postprandial hyperglycemia may have greater pathophysiological consequences than persistent hyperglycemia thus emphasizing the importance of preventing, or rapidly reducing, prandial-induced rises in blood glucose. The recognition that the prediabetic state reflects a postprandial disorder can help identify people with prediabetes and reduce the impact of diabetes-related cardiovascular morbidity and mortality. This review emphasizes strategies designed to minimize the impact of acute effects of hyperglycemia on vascular function. PMID- 18392215 TI - Myocardial infarction. PMID- 18392216 TI - Current challenges and directions in antithrombotic therapy. PMID- 18392217 TI - 2nd international conference on hypertension, lipids, diabetes and stroke prevention. PMID- 18392218 TI - Useful known and unknown views of the father of modern medicine, Hippocrates and his teacher Democritus. AB - Hippocrates is considered to be the father of modern medicine because in his books, which are more than 70. He described in a scientific manner, many diseases and their treatment after detailed observation. He lived about 2400 years ago. He was born in the island of Kos and died at the outskirts of Larissa at the age of 104. Hippocrates taught and wrote under the shade of a big plane tree, its descendant now is believed to be 500 years old, the oldest tree in Europe- platanus orientalis Hippocraticus--with a diameter of 15 meters. Hippocrates saved Athens from a plague epidemic and for that was highly honored by the Athenians. He considered Democritus--the father of the atomic theory--to be his teacher and after visiting him as a physician to look after his health, he accepted no money for this visit. Some of his important aphorisms were: "As to diseases, make a habit of two things -to help or at least to do no harm". Also: "Those by nature over weight, die earlier than the slim.", also, "In the wounds there are miasmata causing disease if entered the body". He used as a pain relief, the abstract from a tree containing what he called "salycasia", like aspirin. He described for the first time epilepsy not as a sacred disease, as was considered at those times, but as a hereditary disease of the brain and added: "Do not cut the temporal place, because spasms shall occur on the opposite area". According to Hippocrates, people on those times had either one or two meals (lunch and dinner). He also suggested: "...little exercise...and walk...do not eat to saturation". Also he declared: "Physician must convert or insert wisdom to medicine and medicine to wisdom". If all scientists followed this aphorism we would have more happiness on earth. PMID- 18392219 TI - The role of PET in head and neck cancer. AB - PET and PET/CT are the procedures of choice for molecular imaging in the head and neck area. The current data of the literature show, that functional imaging with fluorine-18-deoxyglucose ((18)F-FDG) provides the possibility to obtain information about the viability of malignant lesions. The use of hybrid systems, PET/CT, enables physicians to assess both, morphology and function, and achieve a high diagnostic accuracy exceeding 90%. PET with (18)F-FDG is the most sensitive method to detect tumor recurrence. However, false positive results must be considered due to unspecific changes following treatment, especially radiotherapy. The use of quantitative PET scans as well as the application of a second tracer, enhance the capability of PET to assess questionable masses more accurately. Follow up examinations with PET and (18)F-FDG provide data about early changes in the tumor metabolism due to chemotherapeutic treatment. Studies in patients undergoing surgery and radiotherapy demonstrated, that PET with (18)F FDG can be used for the prediction of individual survival. PMID- 18392220 TI - Diagnostic imaging prior to 90Y-ibritumomab tiuxetan (Zevalin) treatment in follicular non-Hodgkin's lymphoma. AB - Add-on treatment of follicular non-Hodgkin's lymphoma (NHL) with yttrium-90 labelled ((90)Y) ibritumomab tiuxetan (Zevalin) has become an efficacious asset in standard treatment concepts of this disease. First-line treatment with Zevalin is currently under way. Whereas in the U.S. and Switzerland a prediagnostic imaging with (111)In-ibritumomab tiuxetan is mandatory, in Europe there is no such prerequisite. It is shown in this article why a prediagnostic imaging or dosimetry is not necessary as an additional mandatory safety measure to confirm the expected biodistribution. PMID- 18392221 TI - Assessment of non-small cell lung cancer viability and necrosis with three radiopharmaceuticals. AB - Assessment of tumor viability and necrosis of non-small cell lung cancer and detection of distant metastases are important for diagnosis, staging, monitoring the response to treatment and planning long-term management. We performed scintigraphy on patients with non-small cell lung cancer to determine the utility of three tumor targeting tracers for diagnosing primary lung cancer, for differentiating viable from necrotic tumor tissue and for detecting distant bone and soft tissue metastases. Our patients were divided into groups. Group A consisted of 27 patients, 25 male and 2 female, mean age 59 years, range from 35 to 72 years. These patients underwent radioimmunoscintigraphy (RIS) using monoclonal antibody against human milk fat globule labeled with technetium-99m ((99m)Tc). Group B consisted of 23 patients, 21 male and 2 female, mean age 56 years, range: 37 to 70 years. Group C consisted of 24 patients, 20 male and 4 female, mean age 58 years, range: 35 to 74 years. Both Groups B and C underwent chest and whole-body scintigraphy with 555 MBq of (99m)Tc-sestamibi ((99m)Tc-S) and 111 MBq of thallium-201 chloride ((201)TlCl), respectively. Tumor to non tumor ratio was calculated. Our findings show that RIS had 52% sensitivity in detecting primary non-small cell lung cancer. In contrast, the sensitivity of (99m)Tc-S and of (201)Tl scintigraphy was 87% and 88%, respectively. High uptake of all three radiopharmaceuticals was found in 6 patients with distant soft tissue and bone tissue metastases and in 1 patient with brain metastasis. Mean tumor to non-tumor ratios were similar: for RIS 1.7+/-0.4, for (99m)Tc-S 1.6+/ 0.3 and for (201)Tl 1.6+/-0.2. IN CONCLUSION: (99m)Tc-S and (201)Tl scintigraphy are superior to RIS for detecting non-small primary lung cancers and potentially clinically useful methods for detecting primary lung cancer as above, as well as bone and soft tissue metastases. PMID- 18392223 TI - Meckel's diverticulum in infants and children; technetium-99m pertechnetate scintigraphy and clinical findings. AB - Meckel's diverticulum (MD) is the most common congenital gastrointestinal anomaly. About 50% of diverticulae contain ectopic gastric mucosa. Gastric mucosal secretions can cause peptic ulceration resulting in pain, bleeding and perforation. Technetium-99m ((99m)Tc) pertechnetate scintigraphy is helpful in diagnosing ectopic gastric mucosa. We have conducted a retrospective analysis of scintigraphic data of 107 pediatric patients, 28 females and 79 males, their age ranging from 5 days to 11 years who referred to us for a query diagnosis of MD. Our results have shown that the most frequent presenting symptom was bleeding per rectum. Twenty-one cases of the 107 were positive for functioning gastric tissue indicating MD (19.62%). Maximum clustering of positive cases was at the age group of 1-2 years (11 cases i.e. 52.38%). The site of the ectopic activity was mainly at the umbilical quatrum in 11 cases (52.4%). Two patients were lost to follow-up and hence surgery could not be performed. The remaining 19 cases were subjected to surgical intervention and 16 were found to be positive for MD. The scintiscan was true positive in 84.2%. Our findings were in agreement with those of other authors. In conclusion, in pediatric patients (99m)Tc-pertechnetate scintigraphy is by 84.2% true positive for gastric mucosa, indicating MD. PMID- 18392222 TI - Four prognostic indices in advanced prostate cancer patients, under palliative androgen deprivation treatment. AB - Prostate cancer (PCa) is the second leading cause of death in men aged 40 years and older ]and prognostic indices are useful in suggesting its proper treatment. The aim of this study was to evaluate the prognostic value of Gleason score (GS), TNM staging system, initial serum prostate specific antigen (PSA) and bone scintigraphy (BS) in patients with PCa under hormonal palliative treatment, in the development and progression of recurrent PCa. Our methods were as follows: Between January 2005 and December 2007, we have studied at the University General Hospital of Alexandroupolis fourty patients of mean age 77+/-7.2 years with advanced PCa under palliative treatment with antiandrogens and luteinizing hormone-releasing hormone analogues. PCa was diagnosed histologically, based on the TNM system after transrectal ultrasonography guided biopsy. The Gleason score assessment was made as described by others. Metastases were confirmed by a positive bone scintigraphy with 925 MBq (99m)Tc-MDP using a tomographic gamma camera, computerized axial tomography or magnetic resonance imagining. Measurements of PSA were conducted by the radioimmunoassay method. We also examined 20 healthy blood donors (median age 45+/-6.1 years) as controls, in order to estimate the cut-off value of PSA. Our results show the following: Thirteen of our patients had 1-6 "hot" spots and 27 had more than 6 "hot" spots in the bone scan. The median Gleason score was 7 (modal Gleason score 3+4). Serum PSA levels were higher in patients with PCa and bone metastases in comparison to those with PCa without bone metastases. Very high values of PSA (more than 50 ng/ml) were found in patients with multiple bone metastases (>6 "hot" spots). In conclusion, our findings demonstrate that the prognostic value of GS (P=0.043), TNM staging (P=0.1410), serum PSA levels (P=0.002) and BS (P=0.0135) when used alone, not always improve the prognosis to hormone indepentent but when combined (P<0.001) increase the prognosis in patients with advanced PCa under hormonal palliative treatment. PMID- 18392224 TI - Is mucociliary transport velocity related to symptoms in chronic rhinosinusitis patients? AB - Mucociliary transport velocity (MTV) measured by rhinoscintigraphy is considered a reliable measure of mucociliary clearance, one of the mechanisms that are distorted in chronic rhinosinusitis (CRS). The aim of this study was to investigate possible correlation of MTV with the severity of patients' symptoms. We have studied thirty-seven CRS patients who completed the 20-item sinonasal outcome test (SNOT-20) form, a reliable measure of health related quality of life and were also examined by rhinoscintigraphy with technetium-99m macroaggregated albumin. Our findings showed that the MTV of our patients ranged from nil to 4.17mm/min with a mean value of 1.74+/-1.055 mm/min. The SNOT-20 scores varied from 0.3 to 3.7 with a mean of 1.6+/-0.922. The correlation between SNOT-20 score and MTV was not statistically significant (Spearman's r=-0.191). In conclusion, although rhinoscintigraphy is widely accepted as a reliable test of mucociliary velocity and clearance, it did not correlate with subjective measures of the severity of CRS. PMID- 18392225 TI - The contribution of Maria Sklodowska-Curie and Pierre Curie to Nuclear and Medical Physics. A hundred and ten years after the discovery of radium. AB - This review aims to commemorate the life, and the accomplishments of Pierre and Marie Curie in Physics and in Medicine. Although they are primarily known for their discoveries of the elements of radium and polonium, which took place two years after the discovery of radioactivity by Henry Becquerel, Pierre's discovery of the piezo-electric phenomenon, his research on crystal symmetry, magnetism and paramagnetic substances, are equally important. With the discovery of the two radioactive elements, Pierre and Marie Curie established the new field of Nuclear Physics. It is not an over-statement to say that their discovery contributed much to our modern way of life. Marie received the Nobel Prize twice and later she became the first woman to become member of the French Academy of Sciences. Today, both Pierre and Marie Curie rest in Pantheon, in Paris. PMID- 18392226 TI - Is sentinel node mapping useful in anorectal melanoma? AB - Anorectal melanoma (AM) is a rare disease and few guidelines have been established regarding its therapeutic management. Wide local excision, or less frequently, abdominoperineal resection is the treatment of choice. We present a female patient with an excised AM who was submitted to our Nuclear Medicine department for sentinel lymph node (SLN) mapping of the area. Under rigid proctoscopy the anal scar received four submucosal injections of technetium-99m sulfur nanocolloid of 29.6 MBq each. Lymphoscintigraphy and intraoperative gamma probe guided detection of the SLNs followed. SLNs were localized in the inguinal basins bilaterally and were negative on histology. A wide local excision followed the above test. Thirty months postoperatively the patient developed distant metastases, underwent radiotherapy and died six months later. Without changing prognosis sentinel lymph node detection, excision and histology were important in sparing the patient a futile inguinal lymph node dissection. PMID- 18392227 TI - Incidental pathologic extracardiac uptake of 99mTc-tetrofosmin in myocardial perfusion imaging. AB - Technetium-99m-tetrofosmin ((99m)Tc-TF) myocardial perfusion studies have incidentally detected various extracardiac abnormalities. The interpretation of these findings may be essential for early diagnosis and treatment of important diseases. We present a rare case of a mediastinal thymoma incidently detected during myocardial perfusion imaging. A 60 year-old woman, with precardiac symptoms of possible myocardial ischemia, underwent a (99m)Tc-TF stress-rest single photon emission tomography test. Intense uptake of the radiotracer in the left paracardiac area, was observed. The computerized tomography and the magnetic resonance imaging tests revealed a mass in the left lower anterior mediastinal area. Biopsy and subsequent histology showed that this mass was a thymoma. PMID- 18392228 TI - The bone scan in disseminated BCGitis. AB - Despite the long history of worldwide use of bacillus Calmette-Guerin (BCG) vaccine, a wide spectrum of adverse reactions has been observed in a small proportion of immunized infants. The most severe complication is disseminated BCGitis, often fatal but extremely rare and considered to be a result of host immunodeficiency. At present, polymerase chain reaction test, CT scan, ultrasound, X-rays and bone marrow aspirations are the investigations used to diagnose this disease. We report a case report of a 6 months old female infant with disseminated BCGitis. This paper aims to highlight the advantages of using bone (99m)Tc-MDP imaging and it's findings in supporting the diagnosis of disseminated BCGitis. PMID- 18392229 TI - White paper of the European Association of Nuclear Medicine (EANM) and the European Society of Radiology (ESR) on multimodality imaging. The opinion of the Department of Nuclear Medicine, University Hospital of Heraklion, Crete, Greece. PMID- 18392230 TI - 18F-FDG-PET, gallium-67 and somatostatin receptor scintigraphy, in ocular MALT lymphoma. PMID- 18392232 TI - Pitfall of 18F-FDG-PET imaging in oncology: herpes zoster with axillary lymphadenopathy. PMID- 18392233 TI - Prostate carcinoma with peripheral metastases after prostatectomy and low levels of serum prostate specific antigen. PMID- 18392234 TI - Advanced multiple myeloma with negative bone marrow biopsy and positive soft tissue lesions in the (18)F-FDG PET/CT scan. PMID- 18392235 TI - [Body composition analysis in obesity: radionuclide and non radionuclide methods]. AB - Body composition (BC) assessment provides important information regarding the absolute or relative amount of bone, lean and fat tissue. Different somatometric techniques have been applied in numerous epidemiological and experimental studies, as well as in every day clinical practice. Traditional techniques for BC analysis include skin fold thickness measurements, radioisotope dilution methods, hydrodensitometry and underwater weighing, while newer techniques include bioelectrical impedance analysis (BIA), air displacement plethysmography (ADP), dual energy X-rays absorptiometry (DEXA), computer tomography and magnetic resonance imaging. In addition, positron emission tomography helped to the functional investigation of adipose tissue, in particular of brown tissue. All these techniques have contributed a lot to the understanding of physiological conditions such as exercise training, menopause and ageing, adolescence health parameters, as well as pathological conditions such as disorders of nutrition, cancer, obesity and diabetes mellitus. In obesity, BC contributed to diagnosis and the pathological impact of visceral adipose tissue. In addition, conditions such as pseudo- or hypermuscular obesity and sarcopenia, which are often observed in various endocrine diseases, were investigated in detail by using such methods. During weight loss, some of these methods were quite accurate in measuring changes in fat and lean mass. Apart from anthropometric measurements, a BC measurement if possible should be included in obesity assessment. Measurements of skin fold thickness combined with BIA are quite sufficient for routine clinical practice. However, in specialized clinics and in research, more sophisticated methods like ADP or DEXA are used. PMID- 18392236 TI - MELD--an imperfect, but thus far the best, solution to the problem of organ allocation. PMID- 18392237 TI - Cancer mortality in Romania, 1955-2004. Digestive sites: esophagus, stomach, colon and rectum, pancreas, liver, gallbladder and biliary tree. AB - AIM: Until recently, gastric cancer was the most frequent digestive neoplasia in our country. Our study presents the first synthesis of data regarding mortality rates from digestive cancers, for a period covering 50 years, in Romania. METHODS: Age-standardized mortality rates /100,000 population, general and/or per gender, concerning six digestive cancers, were identified from the statistics of IARC/OMS (Lyon, France) (years 1955-2002) and of the Ministry of Public Health (Bucharest, Romania) (year 2004). For 2002, incidence and mortality rates per sex from digestive cancers were available and case fatality ratios could be calculated as an approximation of survival rates, as well as sex ratio. RESULTS: Age standardized mortality rates per sex and cancer site registered the following changes: esophageal cancer increased from 2.03/0.62 (M/F) to 2.8/0.5; gastric cancer registered a decrease, from 33.14/18.77 to 17.0/6.6; colorectal cancer increased from 4.65/4.57 to 13.6/9.0; pancreatic cancer increased from 5.50/2.92 to 8.1/4.2 and liver cancer (including peripheric cholangiocarcinoma) increased from 1.77/0.83 to 8.8/3.9. In our population, the case fatality ratio appeared to be better only in colorectal cancer, 0.61 in males and 0.62 in females, respectively. Sex ratio was highest for esophageal cancer (males/females 5.8/1) and lowest for colorectal cancer (1.5/1). CONCLUSIONS: Our study found opposite trends in the mortality rates from digestive cancers, with gastric cancer rates decreasing and the other five digestive cancers increasing. A new hierarchy of digestive cancers has been drawn up, with colorectal cancer as the main cause of death, and gastric cancer in second position, followed by pancreatic, liver, esophageal, and gallbladder and biliary tree cancers. PMID- 18392238 TI - Endoscopic ultrasound-guided fine needle aspiration biopsy in the diagnosis of gastrointestinal stromal tumors of the stomach. A study of 17 cases. AB - AIM: The gastrointestinal stromal tumor (GIST) is an uncommon tumor usually diagnosed by endoscopic biopsy or surgical resection. We evaluated the efficacy and accuracy of endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) biopsy in the diagnosis of GIST. METHOD: Seventeen patients with gastric GIST diagnosed by EUS-guided FNA were included in this study (from 2005 to 2007). A single endosonographer performed all procedures. An attending cytopathologist was present on site to assess specimen adequacy. All tumors were reviewed for EUS, cytomorphologic, histologic and immunohistochemical features. RESULTS: Eleven patients (64.7%) were male and six (35.5%) were female, with a median age of 60.7 years. The clinical indication for EUS-FNA procedure in all patients was the evaluation of submucosal tumor. EUS revealed a solid hypoechoic tumor in all cases with the largest diameter being from 9.5 mm to 70 mm (median diameter 31.9 mm). Cytologic specimen was adequate upon on-site evaluation in all cases, with an average of two passes performed. Spindle cells were present in the cytologic material in all cases and epithelial cells in two cases. Nuclear irregularities and mitoses were not observed. Immunohistochemical (IHC) stain in cell blocks confirmed the c-kit and CD34 positivity in all cases. There were no false negative or false positive findings. CONCLUSIONS: This is the first study of EUS guided FNA procedure in the diagnosis of gastric GIST in Greece. We demonstrated that EUS-FNA provides accurate diagnosis of GIST in combination with IHC reactivity for c-kit, performed in adequate cytology specimens obtained by FNA. PMID- 18392239 TI - Angiotensin II type 1 receptor antagonist improves the prognosis in rats displaying liver cirrhosis induced by a choline-deficient diet. AB - BACKGROUND: Angiotensin II type 1 receptor (AT1) antagonists are known to suppress TGFbeta and lipid peroxidation. An experimental rat model made by feeding rats a choline-deficient diet (CDD) showed severe steatosis, fibrosis and infiltration of inflammatory cells in the liver resembling nonalcoholic steatohepatitis (NASH). NASH causes fibrosis by lipid peroxidation. In this study, we assess whether AT1 antagonists and angiotensin II type 2 receptor (AT2) antagonists can suppress the hepatic fibrosis and lipid peroxidation in CDD rats that lead to the development of NASH. METHODS: Both study groups received subcutaneously aqueous solutions of AT2 antagonist (PD123319 - 1 mg/kg/day) and AT1 antagonist (L158809 - 1 mg/kg/day), respectively, 6 times per week. On day 90, some rats (5 /group) were sacrificed by excision of the liver under anesthesia, in order to assess the hepatic hydroxyproline (HP), malondialdehyde (MDA), total glutathione, superoxide dismutase (SOD) activity and TGFbeta-1. The remaining rats were maintained to observe the survival rate. RESULTS: All CDD rats developed liver cirrhosis. However, the tissue TGF and HP decreased in AT1 antagonist group in comparison with the other two groups. All groups of CDD rats showed strong adipose hyperoxidation. The AT1 antagonist group demonstrated a markedly improved survival rate in comparison to the other two groups. CONCLUSION: Hepatic fibrosis progression in the AT1 antagonist group was slower than that in the other groups. This observation suggests that AT1 antagonists delayed the progression of liver failure, which thus led to an improved survival rate. PMID- 18392240 TI - Postprandial walking but not consumption of alcoholic digestifs or espresso accelerates gastric emptying in healthy volunteers. AB - BACKGROUND: Postprandial consumption of alcoholic beverages with high ethanol concentration (so-called digestifs) is a widespread custom to alleviate dyspeptic symptoms after comprehensive meals. Alcoholic beverages preprandially ingested inhibit gastric emptying rate of solid meals. However, the effect of a postprandial intake has never been studied in a controlled manner. METHODS: In 10 healthy male subjects gastric emptying was repeatedly studied by ultrasonography after the intake of a 576 kcal meal. Immediately after the meal subjects received in a randomized order 40 ml of the following liquids: brandy, herb flavored liqueur, Williams pear brandy, aquavit (each 40 % (v/v) ethanol concentration), espresso, water, 40% (v/v) ethanol and 70% (w/v) glucose. Postprandial satiety, fullness and bloating were determined on a visual analogue scale every 10 minutes. On another occasion subjects received 40 ml of water and walked afterwards slowly (4 km/h) on a treadmill. RESULTS: Gastric half emptying time (t 1/2) of the meal with water was 123 +/- 5 min, while with brandy (119 +/- 9 min), herb flavored liqueur (123 +/- 10 min), aquavit (125 +/- 9 min), Williams pear brandy (126 +/- 6 min) or espresso (125 +/- 9 min) t(1/2) it was not significantly different. Postprandial walking accelerated t(1/2) significantly (107 +/- 5 min, p=0.02). Dyspeptic symptoms were unchanged. Blood ethanol concentrations were under the level of detection (< 5 mg/dl). CONCLUSIONS: Postprandial consumption of alcoholic digestifs did not affect gastric emptying rate of a solid meal nor postprandial dyspeptic complaints. However, postprandial walking accelerated gastric emptying of the meal but this had no effect on dyspeptic symptoms. PMID- 18392241 TI - Clinical and surgical profile and follow up of patients with liver hydatid cyst from an endemic region. AB - AIM: The aim of this study is to present the changes in the treatment of liver hydatid cyst during the last 20 years in our clinic according to literature data. MATERIAL AND METHODS: Clinical, laboratory and operational findings and pre- and postoperative complications of 650 from 700 patients with liver hydatid cysts, examined and treated at Selcuk University Meram Medicine Faculty, General Surgery Department, between 1985-2005, were evaluated in two groups: 1st period (1985 1995) and 2nd period (1995-2005). RESULTS: 436 of the cases were females (67.1%) and 214 (32.9%) males. The mean age of the females was 35 years (ranges 10-73) and of the males 46 years (ranges 12-76). Surgical treatment comprised radical and obliterative conservative techniques in the first period, while non obliterative conservative techniques and percutaneous puncture and aspiration of the cyst, injection of scolex eliminating substance and reaspiration (PAIR) were preferred in the second period. During the follow-up (498 cases were followed for a mean period of 32 [12-72] months), recurrence occurred in 12 in the first period and in 9 in the second period, a total of 21 patients (4.21%). CONCLUSION: We consider that regardless of the surgical treatment used in liver hydatid cyst cases, combination with chemotherapy is the safest and most effective approach. PMID- 18392242 TI - Transthoracic approach (TTA) for subdiaphragmatic liver metastasectomy. AB - BACKGROUND: Hepatic and pulmonary recurrences are major determinants of survival for patients who have undergone curative resection of colorectal carcinoma. In a selected group of patients, resection of metachronous, liver and lung metastases prolongs survival despite the aggressive nature of these lesions. The experience from an exclusive transthoracic, transdiaphragmatic approach (TTA) is limited. We present our experience with metastasectomy in patients with metachronous liver and right lung metastases, in whom an exclusive transthoracic approach was performed. METHODS: Between 2002 and 2007, seven patients with metachronous colorectal liver and right-lung metastases, underwent an exclusive transthoracic approach. There were five men and two women, with a median age of 69 years (range 55 to 78 years). Liver resections performed included segmentectomy of segments VII, VIII, or both. Previous operations, including colon resection, adhesiolysis, ventral hernia repair, or transabdominal segment V resection, were performed in all patients. RESULTS: No peri-operative mortality was documented. Morbidity included pleural effusion (n=3) and post-operative pneumonia (n=1), which responded to conservative management. Median hospital stay was 8 days (range 5-12 days). With a median follow-up of 31 months, one patient died of generalized disease. CONCLUSION: The factors that led to the increase of performances in colonoscopy in our department were the use of proper sedation and analgesia, the permanent internal audit of the maneuver, as well as the motivation of the endoscopist to obtain good results. PMID- 18392243 TI - Postoperative complications linked to pancreaticoduodenectomy. An analysis of pancreatic stump management. AB - AIMS: To analyze the role of different procedures in the management of pancreatic stump according to the incidence of postoperative morbidity derived from the data of a single center surgical population. METHODS: From 1989 to 2005 we performed 76 pancreaticoduodenectomies (PD) and 26 distal pancreatectomies (DP). The surgical reconstruction after PD was as follows: 11 manual non-absorbable stitches closure of the main duct, 24 closures of the main duct with linear stapler, 17 occlusions of the main duct with neoprene glue and 24 duct-to-mucosa anastomosis. RESULTS: In the PD group, the morbidity rate was 60%, caused by: pancreatic leakage in 48% of patients, hemorrhagic complications in 10% following surgical procedure and infectious complications in 15%. After DP we recorded: leakage in 3.9%, haemoperitoneum in 15.4% and no complications in 80.7%. The multivariate analysis showed that the in-hospital mortality was linked to the surgical procedure (PD, p=0.003) and to the following complications: pancreatic leakage (p=0.004), haemoperitoneum (p=0.00045) and infectious complications (p=0.0077). Bleeding complications, biliary anastomosis leakage and infectious complications were consequences of pancreatic leakage (p=0.025, p=0.025 and p=0.025 respectively). CONCLUSION: Manual non-absorbable stitch closure of the main duct and occlusion of the main duct with neoprene glue should be avoided in the reconstructive phase. PMID- 18392244 TI - Combined-modality therapy with sphincter-preserving total mesorectal excision for locally advanced rectal cancer: patient's age and long-term outcome. AB - BACKGROUND: In rectal cancer patients anterior resection with total mesorectal excision (TME) results in good functional outcomes, optimal local control and improved survival. However, patients with locally advanced tumours still have a high risk of oncological relapse and may benefit from neo- or adjuvant therapy. AIM. The purpose of this study was to identify the clinico-pathological features related to poor prognosis after sphincter-saving curative combined-modality therapy for rectal cancer. MATERIAL AND METHOD: Forty-eight consecutive patients with UICC stage II and III rectal carcinoma operated on with TME were studied prospectively. Fifteen patients received neoadjuvant radiotherapy and postoperative chemotherapy; for the remainder combined adjuvant chemoradiotherapy was given. RESULTS: Five-year cancer-specific survival rate was 49%. Patient's gender, tumour location, grade, mucinous histology, direct tumour spread, type of growth margin and lymphocytic tumour infiltration were revealed to be factors without statistical importance. Only the positive nodal status (31.6 +/- 11.0 vs 61.5 +/- 9.7) and the patient's advanced age (38.5 +/- 9.7 vs 63.2 +/- 11.4) were significantly related to decreased survival rate (p=0.038 and 0.048, respectively). In multivariate analysis both parameters independently influenced poor prognosis (p=0.045 and 0.038; Relative Risk 2.26 and 2.13; Odds Ratio 4.21 and 1.07, respectively). CONCLUSIONS: An elderly patient's age seems to be an independent prognostic factor associated with poor survival after curative treatment for locally advanced rectal cancer even when non-cancer causes of death are excluded. Thus, for older patients adjuvant therapy should be an integral part of treatment with the careful benefit-toxicity analysis. PMID- 18392245 TI - Recurrence pattern of fistula-in-ano in a Chinese population. AB - BACKGROUND AND AIMS: Fistula-in-ano is a common colorectal complaint. Despite of the advancement in preoperative imaging and surgical techniques, recurrence is not uncommon in this disease entity. We aimed to determine the recurrence pattern and predictors of FIA in Chinese population. METHODS: SETTING: single regional hospital serving a 300,000 population. DESIGN AND PARTICIPANTS: A systematic retrospective review of inpatient and follow up records and operation records in patients with diagnosis of fistula-in-ano (ICD code - 565.1) from January 2001 to December 2004 was performed. INTERVENTION: Surgery for fistula-in-ano. RESULTS: 135 out of 137 Chinese patients received anal fistula operations. 14 patients (10.4%) had high type anal fistulas and 27 patients (20%) had perianal sinus. The most common operation was combined fistulotomy-fistulectomy (78 patients, 57.8%). 18 patients (13.3%) had recurrence with a median time to recurrence of 7.5 months. Six factors including: 1) positive history of perianal abscess, 2) previous perianal operation, 3) complex fistula, 4) perianal sinus, 5) absence of an internal opening, 6) surgical procedure of sinus tract excision were significantly associated with recurrence in univariate analysis. Sinus tract excision was the only independent factor to predict recurrence in logistic regression (p=0.002, 95%CI=1.29-3.27). CONCLUSION: Fistula-in-ano carried a significant risk of recurrence in perianal sinus with sinus tract excision performed. No difference was found between Chinese and Caucasian in recurrence pattern of fistula-in-ano. PMID- 18392246 TI - Principles and applications of gene therapy in colon cancer. AB - Colorectal cancer is one of the leading causes of mortality and morbidity. Mutations and aberration of some of the genes may lead to colon cancer. With advancing knowledge, more and more defective genes can be identified. Theoretically, correction of these defective genes, selective overexpression of certain genes may lead to not only prevention of cancer development but also regression of existing cancer. Carcinogenesis is a multistep process and more than one gene may be altered. At present, the success of gene therapy as a stand alone therapy is limited by poor expression and long-term non-expression of transferred genes, immunological effect on the viral vectors, viraemia, leukaemia and occasional deaths. Further experiments and trials are underway to modify the vectors and genes to make sure gene therapy is safe, effective and long-lasting before it can be used in day-to-day practice. PMID- 18392247 TI - Updating the management of rectal cancer. AB - In the last 25 years rectal cancer has changed from a surgically managed disease into a multidisciplinary treatment model. Accurate staging has a critical role in the decision-making process of patients with rectal cancer. The four most commonly used imaging modalities in the pretherapeutical staging include endoscopic ultrasound, computerized tomography, magnetic resonance imaging and positron emission tomography. Locoregional tumor control in rectal cancer surgery has improved significantly over the last 15 years, after the introduction of total mesorectal excision (TME), which leads to the complete removal of the intact mesorectum including the lymphatics, lymph nodes, nerves, and vascular supply. At the present time, given the improved local control, acute and long term toxicity profile, and sphincter preservation rate, patients who require combined modality therapy should receive concomitant radiochemotherapy preoperatively. Recently, the novel 'targeted' therapies have been incorporated into a multidisciplinary approach for rectal cancer. PMID- 18392248 TI - QUIZ HQ 42. A 45-year-old woman with rapid progressive abdominal distension. PMID- 18392249 TI - Intestinal pseudo-obstruction--a rare condition with heterogeneous etiology and unpredictable outcome. A case report. AB - Intestinal pseudo-obstruction (IPO), either acute or chronic, is a condition including features of intestinal ileus in the absence of mechanical obstruction. We present such a rare case of idiopathic IPO in a 53-year old male patient in whom recurrent episodes of pseudo-obstruction were successfully resolved by anticholinesterase agents, motilin agonists or colonic decompression. However, subsequently total colectomy was required. Huge colonic dilatation was identified intraoperatively, while histology evidenced a neuropathic variant of chronic intestinal pseudoobstruction. IPO is a condition in which conservative treatment usually fails. Total colectomy with ileoanal pouch was the solution in our patient. PMID- 18392250 TI - Management of Boerhaave's syndrome: report of three cases. AB - From 2000 to 2005, three patients with Boerhaave's syndrome were successfully managed in our Department. Two of them received the appropriate treatment belatedly, with primary closure and bolstering tissue wrap. One of them required further intervention with a cervical esophagostomy and exclusion of the perforated esophagus. The third patient with an esophageal perforation related disorder, was managed with surgical exploration and drainage alone. Primary suturing of the esophagus should be performed only in patients with an early perforation. In cases of prolonged delay between rupture and diagnosis, esophageal resection with cervical esophagostomy and gastrostomy is advocated as the safest therapy. PMID- 18392251 TI - QUIZ HQ 43. An unusual cause of obstructive jaundice. PMID- 18392252 TI - Bouveret syndrome associated with acute gangrenous cholecystitis. AB - An 89-year-old patient was hospitalized with signs of acute lithiasic cholecystitis and gastric emptying failure. The decision for surgery was taken and a subhepatic block was evidenced, caused by a perforated gangrenous cholecystitis with pericholecystic abscess, a cholecysto-antroduodenal fistula with two gallstones, 9/5 and 4/3 cm in size, impacted in the duodenum. It was necessary to perform an Y-en-Roux antroduodenojejunal anastomosis because an antroduodenal parietal defect resulted after the removal of the gangrenous gallbladder. The immediate and long term postoperative evolution in terms of anastomosis functionality was good. PMID- 18392253 TI - Retractile mesenteritis presenting with malabsorption syndrome. Successful treatment with oral pentoxifylline. AB - Retractile mesenteritis is a rare benign inflammatory disease of the mesentery. Computed tomographic findings usually suggest the diagnosis, which is confirmed by surgical biopsies. Conservative treatment is empirical, based on corticosteroids, colchicine, immunosuppressive agents and progesterone. Surgical resection is sometimes attempted for definitive therapy, although the surgical approach is often limited. This report describes a 62-year old man with histologically proven retractile mesenteritis presenting with malabsorbtion syndrome, who presented pulmonary tuberculosis after initial therapy with corticosteroids. He was subsequently treated with oral pentoxifylline (800 mg/day), with substantial clinical and radiological improvement. PMID- 18392254 TI - Confocal endomicroscopy for in vivo microscopic analysis of upper gastrointestinal tract premalignant and malignant lesions. AB - Confocal LASER endomicroscopy (CLE) is a new endoscopic technique which allows subsurface in vivo microscopic analysis during ongoing endoscopy, using systemically or topically administered fluorescent agents. It allows targeted biopsies to be taken, potentially improving the diagnostic rate in certain gastrointestinal diseases. Worldwide experience with CLE for upper gastrointestinal malignant and premalignant lesions is still reduced. Potential clinical applications are presented, including diagnosis of NERD, Barrett's esophagus, atrophic gatritis, gastric intestinal metaplasia and dysplasia, gastric adenomatous or hyperplastic polyps, gastric cancer. PMID- 18392255 TI - Splanchnic artery aneurysms: the diagnostic contribution of ultrasonography in correlation with other imaging methods. AB - Splanchnic artery aneurysms are localized vascular dilatations having a progressive evolution and leading towards severe complications (depletive rupture, distal emboli, organ ischemia). Treatment can be efficient when the condition is detected in time. The imaging techniques are highly efficient when it comes to detecting and locating splanchnic aneurysms. Each method has its own features. By knowing them, there is a better chance to track down the disease at an early stage. PMID- 18392256 TI - QUIZ HQ 44. Unusual cause of breathlessness. PMID- 18392257 TI - Remarkably elevated serum levels of carbohydrate antigen 19-9 in cystic duct and common bile duct lithiasis. PMID- 18392258 TI - First intoxication with freshly picked Amanita phalloides in winter time in central Europe. PMID- 18392259 TI - Cerebral sinus thrombosis in a patient with ulcerative colitis. PMID- 18392260 TI - Extended poststrangulation ischemic jejunal stricture. PMID- 18392261 TI - Prevalence of upper and lower gastrointestinal symptoms in Greek patients with type 2 diabetes mellitus. PMID- 18392262 TI - A case of pyloric-type gallbladder adenoma. PMID- 18392269 TI - Environment: challenges and opportunities. PMID- 18392270 TI - Nanotechnology, risk and the environment: a review. AB - Nanotechnologies are already interacting with the environment. Scientists and engineers are manipulating matter at the nanoscale, and these nanoscale processes and products are being used by industry in commercially available products. These products are either applied directly to the environment or end up in the environment through indirect pathways. This review examines the state of current environmental risk assessment of nanotechnologies. Nanotechnology is described generally, then both the possible benefits of nanotechnology and the risks are reviewed in a traditional way. Subsequently, a philosophical criticism of the traditional way of looking at risks is offered. PMID- 18392271 TI - Use of mass spectrometry for determining microbial toxins in indoor environments. AB - We know little about the health consequences of human exposure to microorganisms in the environment. This may largely be due to the lack of universally accepted, sensitive, reproducible, non-discriminatory, and health relevant methods for characterizing the exposure. In this review, some of the different mass spectrometry methods that have been developed for measuring endotoxin and peptidoglycan from bacteria and mycotoxins from moulds in samples from-mainly damp-indoor environments are discussed. PMID- 18392272 TI - Assessment of diesel particulate matter exposure in the workplace: freight terminals. AB - A large study has been undertaken to assess the exposure to diesel exhaust within diesel trucking terminals. A critical component of this assessment is an analysis of the variation in carbonaceous particulate matter (PM) across trucking terminal locations; consistency in the primary sources can be effectively tracked by analyzing trends in elemental carbon (EC) and organic molecular marker concentrations. Ambient samples were collected at yard, dock and repair shop work stations in 7 terminals in the USA and 1 in Mexico. Concentrations of EC ranged from 0.2 to 12 microg m(-3) among the terminals, which corresponds to the range seen in the concentration of summed hopanes (0.5 to 20.5 ng m(-3)). However, when chemical mass balance (CMB) source apportionment results were presented as percent contribution to organic carbon (OC) concentrations, the contribution of mobile sources to OC are similar among the terminals in different cities. The average mobile source percent contribution to OC was 75.3 +/- 17.1% for truck repair shops, 65.4 +/- 20.4% for the docks and 38.4 +/- 9.5% for the terminal yard samples. A relatively consistent mobile source impact was present at all the terminals only when considering percentage of total OC concentrations, not in terms of absolute concentrations. PMID- 18392273 TI - Magnesium hydroxide bulk and colloid-associated 152Eu in an alkaline environment: colloid characterisation and sorption properties in the presence and absence of carbonate. AB - The distribution of 152Eu between magnesium hydroxide bulk, colloids and solution has been assessed under alkaline conditions, such as those in nuclear fuel storage ponds. The colloidal phase has been characterised by two complementary methods: coupled ultrafiltration-ICP-AES and scanning electron microscopy. The quantity and the size distribution of the colloidal phase is strongly ionic strength-dependent. A decrease of the quantity of colloids, in particular the larger size ranges, has been observed with increasing ionic strength. Small colloids (1 kDa-10 kDa fraction) are predominant at all ionic strengths. The morphology of colloids, observed by field-emission gun scanning electron microscopy, appears to change from hexagonal prismatic (characteristic to the mineral) to spherical (energetically more favourable) as size decreases. The distribution of 152Eu between the solid and liquid/colloidal phases has been investigated at carbonate concentrations ranging from 0 to 10(-2) M by coupled ultrafiltration and gamma-spectrometry. Mg(OH)2 bulk appears to be a very strong sorbent for 152Eu, since complete sorption onto the bulk happens for carbonate concentrations as high as 10(-3) M. Scavenging of 152Eu by Mg(OH)2 colloids is negligible in the presence of Mg(OH)2 bulk. The distribution of 152Eu between liquid and colloidal phases has been investigated in the absence of bulk at various carbonate concentrations. A significant uptake of 152Eu by the colloids in solution has been observed, which decreases with increasing carbonate concentration. 152Eu appears to be mainly associated to the smallest colloids (1 kDa-10 kDa fraction). There is a strong correlation between the sorption properties and the surface area of the colloids. PMID- 18392274 TI - Screening volatile organic compounds (VOCs) emissions from five marine phytoplankton species by head space gas chromatography/mass spectrometry (HS GC/MS). AB - Five marine cosmopolitan phytoplankton species namely; Calcidiscus leptoporus, Emiliania huxleyi, Phaeodactylum tricornutum, Chaetoceros neogracilis and Dunaliella tertiolecta were screened for emissions of selected VOCs using head space gas chromatography/mass spectrometry (HS-GC/MS) in single ion mode. The VOCs investigated included isoprene and various halogenated compounds. Among the different algae groups, the two diatoms Ch. neogracilis and P. tricornutum were the strongest emitters of methyl bromide (CH3Br), and Ch. neogracilis was the strongest emitter of isoprene. Furthermore, we present evidence that several chlorinated organic compounds, normally considered as anthropogenic, can be produced from marine phytoplankton (namely chloroform, dichloromethane, trichloroethylene, tetrachloroethylene, chlorobenzene and dichlorobenzene). PMID- 18392275 TI - Passive aerosol sampler for particle concentrations and size distributions. AB - This research evaluated the UNC passive aerosol sampler as a tool to measure particle mass concentrations and size distributions. The exposure scenario represented high concentrations and exposure periods of a few hours. Mass concentrations measured with the passive sampler were compared to concentrations measured using both a dichotomous sampler and an aerodynamic particle sizer (APS). In addition, the size distributions measured with the passive sampler were compared to those measured using the APS. Mass concentrations measured using the dichotomous sampler and the APS agreed well. The passive sampler tracked, but tended to overestimate, mass concentrations measured by the other two instruments. Size distributions measured with the passive sampler followed the general pattern of those measured using the APS. Overall, the passive sampler demonstrated both its utility and its limitations in these tests. The concentration measurements and size distributions found using passive samplers were more variable than those of the other instruments, but generally followed the data taken using the other methods. The advantages of low cost and ease of use offset the limitations in data quality with the passive sampler; these advantages are particularly welcome for sampling situations where aerosol properties vary over space or time. PMID- 18392276 TI - Development and application of quantitative methods for monitoring dermal and inhalation exposure to propiconazole. AB - Quantitative methods to measure dermal and inhalation exposure to the fungicide propiconazole were developed in the laboratory and applied in the occupational exposure setting for monitoring five farm workers' exposure during pesticide preparation and application to peach crops. Dermal exposure was measured with tape-strips applied to the skin, and the amount of propiconazole was normalized to keratin content in the tape-strip. Inhalation exposure was measured with an OVS tube placed in the worker's breathing-zone during pesticide handling. Samples were analyzed by GC-MS in EI+ mode (limit of detection 6 pg microl(-1)). Dermal exposure ranged from non-detectable to 32.1 +/- 22.6 ng per microg keratin while breathing-zone concentrations varied from 0.2 to 2.2 microg m(-3). A positive correlation was observed between breathing-zone concentrations and ambient air temperature (r2 = 0.87, p < 0.01). Breathing-zone concentrations did not correlate with dermal exposure levels (r2 = 0.11, p = 0.52). Propiconazole levels were below limit of detection when rubber gloves, coveralls, and full-face mask were used. The total-body propiconazole dose, determined for each worker by summing the estimated dermal dose and inhalation dose, ranged from 0.01 to 12 microg per kg body weight per day. Our results show that tape-stripping of the skin and the OVS can be effectively utilized to measure dermal and inhalation exposure to propiconazole, respectively, and that the dermal route of exposure contributed substantially more to the total dose than the inhalation route. PMID- 18392277 TI - Tape-stripping as a method for measuring dermal exposure to resin acids during wood pellet production. AB - The purpose of this study was to develop a sensitive and specific method for quantifying dermal exposure to the resin acids 7-oxodehydroabietic acid (7-OXO), dehydroabietic acid (DHAA), abietic acid (AA), and pimaric acid (PA). In addition the method was evaluated in occupational settings during production of wood pellets. Tape-strips were spiked with the substances to evaluate the recovery of the acids from the tape. The removal efficiency of the tape was assessed by tape stripping a specified area on a glass plate spiked with resin acids. The recovery of the acids from human skin in vivo was evaluated by applying acids in methanol onto the skin of volunteers. Occupational dermal exposure to the resin acids was assessed by tape-stripping the skin of workers involved in the production of wood pellets. The resin acids were analyzed by liquid chromatography mass spectrometry (LC-MS). The limit of detection was 15 pg (7-OXO), 150 pg (DHAA), 285 pg (AA) and 471 pg (PA) per injection. The recovery from spiked tapes was in general 100%. The removal efficiency of the tape was 48-101%. Recovery tests from human skin in vivo showed a mean recovery of 27%. Quantifiable amounts of resin acids were observed on four different skin areas with an increase in exposure during a work shift. This study shows that occupational dermal exposure to resin acids can be assessed by tape-stripping and quantified by LC-MS. PMID- 18392278 TI - Multi-residue determination of pharmaceuticals in sludge and sludge enriched soils using pressurized liquid extraction, solid phase extraction and liquid chromatography with tandem mass spectrometry. AB - An analytical method to determine a selection of 27 frequently prescribed and consumed pharmaceuticals in biosolid enriched soils and digested sludges is presented. Using a combination of pressurized liquid extraction, solid phase extraction and liquid chromatography with tandem mass spectrometry, it was possible to detect all analytes in each sample type at the low-sub ng g(-1) level. Solid phase extraction efficiencies were compared for 6 different sorbent types and it was found that Waters Oasis HLB cartridges offered enhanced selectivities with 20 analytes showing final method recoveries > or =60% in both soils and digested sludges. The method was validated for linearity, range, precision and limits of detection in both sample matrices. All analytes were then determined in sludge enriched soils as well as the precursor thermally dried sludge fertilizer produced from a primary wastewater treatment plant. Levels of the antibacterial agent triclosan were found to exceed 20 microg g(-1) in digested sludge and 5 microg g(-1) in thermally dried sludge cake. Significant traces of carbamazepine and warfarin were also detected in the above samples. PMID- 18392279 TI - Recovery of Francisella tularensis from soil samples by filtration and detection by real-time PCR and cELISA. AB - The aim of this study was to develop a specific and highly sensitive method able to detect very low concentrations of Francisella tularensis in soil samples by real-time PCR (qPCR) with SYBR Green I. tul4 gene, which encodes the 17-kDa protein (TUL4) in F. tularensis strains, was amplified using a LightCycler (LC) device. We achieved a detection limit of 0.69 fg of genomic DNA from F. tularensis subp. holarctica live vaccine strain (LVS), corresponding to a value less than 3.4 genome equivalents per reaction. The qPCR was shown to be specific, highly sensitive and reproducible. In addition, we evaluated 2 new methods for recovering bacteria from soil based on 1-step filtration using glass fiber filters and PVDF filters. These filtration methods enabled us to recover F. tularensis efficiently from soil samples. As few as 50 CFU per 0.5 g of soil were detected by qPCR. Capture enzyme-linked immunosorbent assay (cELISA) allowed us to detect and quantify the amount of bacteria recovered from soil by an immunological method. Although qPCR was more sensitive than cELISA, we did not observe substantial differences in the amount of bacteria quantified by both methods. PMID- 18392280 TI - Spatial and temporal variations in inhalable CuZnPb aerosols within the Mexico City pollution plume. AB - We report on the CuPbZn content of PM10 and PM2.5 samples collected from three sites (urban T0, suburban T1 and rural T2) during the Mexico City MILAGRO campaign of March 2006. Daytime city centre concentrations of summation operator CuZnPb(PM10) were much higher (T0 > 450 ng m(-3)) than at the suburban site (T1 < 200 ng m(-3)). Rural site (T2) summation operator CuZnPb(PM10) concentrations exceeded 50 ng m(-3) when influenced by the megacity plume but dropped to 10 ng m(-3) during clean northerly winds. Nocturnal metal concentrations more than doubled at T0, as pollutants became trapped in the nightly inversion layer, but decreased at the rural site. Transient spikes in concentrations of different metals, e.g. a "copper event" at T0 (CuPM10 281 ng m(-3)) and "zinc event" at T1 (ZnPM10 1481 ng m(-3)) on the night of March 7-8, demonstrate how industrial pollution sources produce localised chemical inhomogeneities in the city atmosphere. Most metal aerosols are <2.5 microm and SEM study demonstrates the dominance of Fe, Ti, Ba, Cu, Pb and Zn (and lesser Sn, Mo, Sb, W, Ni, V, As, Bi) in metalliferous particles that have shapes including spherical condensates, efflorescent CuZnClS particles, cindery Zn, and Cu wire. Metal aerosol concentrations do not change in concert with PM10 mass, which is more influenced by wind resuspension than industrial emissions. Metalliferous particles can induce cell damage, and PM composition is probably more important than PM mass, with respect to negative health effects, so that better monitoring and control of industrial emissions would likely produce significant improvements in air quality. PMID- 18392282 TI - Chris Polge, CBE FRS (16 August 1926 - 17 August 2006): a personal appreciation. PMID- 18392281 TI - Sampling and analytical methodology development for the determination of primary and secondary low molecular weight amines in ambient air. AB - In this project, a sampling device and an analytical method have been developed to simultaneously analyse the most frequently found low molecular weight amines, including aliphatic, aromatic and alcohol amines. These amines are diethanolamine, ethanolamine, methylamine, isopropylamine, morpholine, dimethylamine, and aniline. A sampling device was developed using a 37 mm cassette with glass fibre filters impregnated with sulfuric acid. Immediately after sampling, the filter was transferred to vials containing a solution of dansyl chloride. Dansyl chloride was used for derivatisation because it forms aromatic sulfonamides that are fluorescent and easy to protonate for MS detection. The effect of using an internal standard made with the dansylated derivative of 1-(2-methoxyphenyl)piperazine (MOPIP) on the uncertainty and efficiency of the method was also evaluated. This internal standard was spiked directly onto filters. The coupling of HPLC/ESI-MS was used for the simultaneous analysis of all the derivatives. This method showed detection limits of about 0.03 microg mL(-1) to 0.3 microg mL(-1) of amine with an average expanded uncertainty of 3% to 6% depending on the amine. The methodology recoveries are close to 100% for all the amines, and the overall estimated expanded uncertainties vary between 10% and 13% depending on the amine. This new strategy will be useful in evaluating workplace air since a unique sampling system will be used, independent of the amine to be quantified. PMID- 18392283 TI - Determination of genetic stability in surviving apple shoots following cryopreservation by vitrification. AB - In vitro-grown apple shoot tips were successfully cryopreserved by vitrification, with an average survival and shoot formation of 80 percent and 76 percent respectively. Surviving shoots showed the same rate and regrowth patterns as those of non-treated controls. No significant differences (P < 0.05) were observed in morphological characteristics, including shoot length, leaf shape, leaf width/length ratio and root length, between the control and cryopreserved shoots. No different microsatellite alleles and ISSR fragments were detected between control and cryopreserved shoots using twelve pairs of microsatellites and eleven ISSR primers. These results show that cryopreservation using vitrification is a practical method for the long-term storage of apple germplasm. PMID- 18392284 TI - Deployment of the encapsulation-dehydration protocol to cryopreserve microalgae held at the Sammlung von Algenkulturen, Universitat Gottingen, Germany. AB - Encapsulation-dehydration was applied to cryopreserve 14 diverse algal strains, representing eukaryotic terrestrial microalgae; of these 12 survived to form cell colonies after recovery from cryostorage. Surviving algae had varying degrees of tolerance to osmotic dehydration and desiccation in this vitrification-based cryoprotective strategy. The extent of algal regrowth was affected by the mode of desiccation (silica gel or air-flow), the duration of evaporative desiccation and exposure to light during early recovery phase. This paper: (i) demonstrates the versatility of the encapsulation/dehydration method to cryopreserve diverse microalgae; (ii) confirms the successful transfer of this cryostorage technology to the Culture Collection of Algae at Gottingen University (SAG); and (iii) recommends encapsulation/dehydration as a feasible alternative to controlled rate cooling for preserving algae held in international culture collections. PMID- 18392285 TI - Deployment of the encapsulation-dehydration protocol to cryopreserve diverse microalgae held at the Institute of Soil Biology, Academy of Sciences of the Czech Republic. AB - Twenty-seven strains of soil algae isolated from highly diverse provenances and habitats were assessed for their capacity to withstand cryopreservation using encapsulation/dehydration. Survival was assessed following the release of algae from alginate beads treated with sodium hexametaphosphate and regrowth was assessed using NAJA Image Analysis. Regrowth occurred in 19 strains, with > 50% survival being observed in 15. Algal tolerance to osmotic dehydration and evaporative desiccation was critical to the success of the method. Recovery in five out of the remaining eight recalcitrant strains was enhanced by substituting sorbitol for the osmotic pretreatment or by combining encapsulation with two-step controlled rate cooling. PMID- 18392286 TI - Deployment of the encapsulation-dehydration protocol to cryopreserve polar microalgae held at the Czech Republic Academy of Sciences Institute of Botany. AB - Polar isolates of four chlorococcal microalgae originating from the Arctic and Antarctica withstand cryopreservation using encapsulation-dehydration. Viability assessments, which initially used chloroplhyll fluorescence (Kautsky) induction kinetics, revealed that all strains suffered photosynthetic impairment during early post-cryopreservation recovery. This cryoinjury was reversible, as indicated by cell regrowth in three of the four strains. Lack of growth in the fourth isolate was due to contaminating bacteria rather than cryogenic factors. PMID- 18392287 TI - Cubic ice can be formed directly in the water phase of vitrified aqueous solutions. AB - This study addresses the possibility that ice-Ic may form spontaneously and directly during the cooling of previously freeze-concentrated aqueous solutions, at temperatures where the residual unfrozen water is likely to exist in small, isolated domains. It is shown that on cooling a dilute aqueous glucose solution, ice-Ih crystallises initially, but when the solution has been freeze-concentrated to ca.10 mol %, further cooling results in the slow nucleation and growth of ice Ic in the residual water domains, rather than on the pre-existing ice-Ih crystal surfaces. PMID- 18392288 TI - Increase in the contribution of transamination to the respiration of mitochondria during arousal. AB - The involvement of transamination in the respiration of liver mitochondria in ground squirrels during hibernation and arousal has been studied. It was shown by HPLC that, in the presence of glutamate and malate, the formation of alpha ketoglutarate (KGL), a transamination marker, and fumarate, a product of succinate oxidation, takes place. During arousal, the formation of KGL increased fourfold, and the respiration sensitive to the inhibitor of aspartate transaminase aminooxyacetate (AOA) increased threefold. The function of transamination upon arousal is related to the elimination of oxaloacetate, an endogenous inhibitor of succinate dehydrogenase. In addition, being more resistant to oxidative stress than oxidation, transamination is probably involved in the antioxidant defense required during the rapid rise of body temperature upon arousal. Our experiments showed an increase in the concentration of malonic dialdehyde (MDA), an end product of lipid peroxidation, in liver mitochondria in this state, which can reduce the activity of the enzymes of the tricarboxylic acids cycle. Under these conditions, the transamination contributes to the maintenance of a high respiration rate necessary for arousal. PMID- 18392289 TI - Cryobiology, wildlife conservation and reality. AB - Conservation is about protecting and nurturing species so that they can survive, not only now but also into the future. Ideally this means protecting genetically diverse populations and not simply breeding a few individuals. In principle, cryobiology offers the means to help maintain genetic diversity by storing genetically important germplasm that could reinvigorate populations in the future. Unfortunately the technical problems associated with this ideal goal still provide a major barrier to the practical use of cryopreservation technology. Sometimes these are technical problems with the cryobiology, but lack of basic biological information about unusual species, coupled with difficulties in obtaining such information, means that progress will be possible with only a few species that are subject to intensive scrutiny. The opportunities nevertheless exist for cryobiologists and reproductive biologists to make useful and global contributions to species conservation. I argue here that there are often two mutually suspicious groups of biologists, who do not interact or even understand each others goals. If conservation biologists and biotechnologists were more prepared to join forces and share their expertise, there would be much improved prospects for achieving lasting success in the conservation of a small, but well targeted, number of threatened species. PMID- 18392290 TI - Cryopreservation for plant genebanks - a matter between high expectations and cautious reservation. AB - Cryopreservation is the best method of storing germplasm efficiently and safely, particularly for the maintenance of vegetatively propagated material. In IPK cryopreservation is used for potato, garlic, mint and yam. IPK collaborates with other cryobanks and research groups (ECPGR, COST, EURALLIVEG) and finds considerable differences in the adoption of cryopreservation between crops and their host institutes, depending on crop, local and historical circumstances. A better understanding of the long-term benefits of cryopreservation and its further integration into general genebank management is therefore needed. Recommended approaches include: comparative validation of methods between different laboratories, detailed comparisons of crop-based methods, economical analyses, efficient integration strategies of cryobanks by genebanks; including safe duplication of cryopreserved resources for the limitation of risk of loss Importantly, there has been recent progress in the development of quality management systems. Cryopreservation is, however, characterized by high expectations. Therefore, to ensure its sustainable and practicable use, basic knowledge of storage protocols must be combined with increased awareness of the rationales required to validate, implement and apply cryobanking technologies in working genebanks. PMID- 18392291 TI - The impact of OECD best practice on the validation of cryopreservation techniques for microorganisms. AB - The Organisation for Economic Co-operation and Development (OECD) Biological Resource Centre Initiative (BRC) was established after the 1998 Working Party on Biotechnology endorsed a proposal by Japan to examine support for Biological Resource Centres (BRCs) as a key element of the scientific and technological infrastructure for the life sciences and biotechnology. As part of this Best Practice Guidelines for the operation of Biological Resource Centres (BRCs) were published. Cryopreservation is widely used in BRC's and is seen as the method of choice for the preservation of most organism groups. This paper reviews the developments of BRC standards, how they are applied, current practices in cryopreservation and methods for validating the success of cryopreservation methodology. PMID- 18392292 TI - Embryonic stem cells and Parkinson's disease: cell transplantation to cell therapy. PMID- 18392293 TI - Forty-one cervicofacial vascular anomalies and their surgical treatment- retrospection and review. AB - INTRODUCTION: Haemangiomas in children usually involute spontaneously and surgical treatment is exceptional. Vascular malformations do not regress spontaneously and resection may become necessary. We present a series of surgically treated face and neck vascular anomalies during a 9-year period, assessing the epidemiology, presenting signs and symptoms, diagnostic modalities, indications for surgery, treatment methods and clinical outcome post-treatment. MATERIALS AND METHODS: The medical and pathological records of all patients with cervicofacial vascular anomalies treated surgically at our department from 1997 to 2005 were retrospectively reviewed in relation to current evidence. RESULTS: Forty-one patients were identified. Of these, 9 patients had haemangiomas and the remaining 32 had a variety of vascular malformations. Cervicofacial vascular anomalies were most commonly located at the lip. Atypical looking vascular anomalies like masseteric intramuscular haemangiomas and parotid malformations were diagnostic problems. All 41 had surgical excision of their vascular anomalies for troubling symptoms, cosmesis or diagnostic purpose. For cervicofacial arteriovenous malformations, 28% were classified as Schobinger stage I, 50% stage II, and the remainder stage III. Combined embolisation resection was used to treat 6 arteriovenous malformations (stage II to III) and of these, 3 required flap reconstruction. CONCLUSIONS: Accurate diagnosis distinguishing between cervicofacial haemangiomas and vascular malformations is key to best treatment. The diagnosis can usually be made by history and physical examination aided by early magnetic resonance imaging (MRI). Although cervicofacial haemangiomas can be managed conservatively or with medical therapy, surgery is indicated for preventing psychological distress and in cases of chronic aesthetic alteration resulting from partial regression. Aesthetic concerns and prevention of psychosocial distress point to early excision of venous malformation as the treatment of choice. Lymphatic malformations are best treated by excision. Outcome after excision of localised cervicofacial haemangiomas and low-flow vascular malformations is excellent. Large extensive low-flow malformations as well as those located at the lips may require multiple procedures including reconstruction; patients should be informed that the outcome is generally not as good. Combined embolisation-resection is definitive treatment for arteriovenous malformations and flap reconstruction may prevent their recurrence. Tissue expansion is a useful reconstructive tool after the excision of large vascular anomalies. PMID- 18392294 TI - Charity colonoscopy event to commemorate the 185th anniversary of Singapore General Hospital. AB - INTRODUCTION: Colorectal cancer is now the cancer with the highest incidence in Singapore. However, the overall mortality rate is still about 50% because the majority of the patients present at a late stage of disease. A charity event of screening colonoscopy was offered to the public in conjunction with the 185th anniversary of Singapore General Hospital. The aim of this event was to raise awareness about early detection of colorectal cancer and the safety of colonoscopy. MATERIALS AND METHODS: We conducted a one-off free screening event for colorectal cancer using colonoscopy. Four hundred and ninety individuals responded to a multimedia advertisement for the event. Of these, 220 individuals were selected for the screening based on National Guidelines for colorectal cancer screening and financial status. RESULTS: One hundred and fifty-two individuals turned up for the colonoscopy. The median age was 55 years (range, 22 to 82), with 84 males. Significant pathology was found in 33% of the individuals (n = 51). Colorectal polyps were detected in 34 individuals (22%). A total of 45 polyps were removed, with 20 hyperplastic polyps and 25 adenomas. Eight out of 25 adenomas were located proximal to the splenic flexure. Rectal cancer was diagnosed in 1 individual (0.6%). One individual had a large dysplastic rectosigmoid ulcer and refused further intervention. There were no significant complications from any of the colonoscopies. CONCLUSIONS: Colonoscopy is an invaluable screening modality as it has a high pick-up rate for colorectal polyp and cancer in an asymptomatic population. It is also proven to be safe in our study. It has the added advantage over flexible sigmoidoscopy of detecting a significant number of proximal lesions. Also, therapeutic polypectomy can be performed in the same setting. PMID- 18392295 TI - Review of the management outcome of slipped capital femoral epiphysis and the role of prophylactic contra-lateral pinning re-examined. AB - INTRODUCTION: Slipped capital femoral epiphysis is the most common hip problem in the adolescent age group. It can involve both hips, presenting itself bilaterally at first presentation or sequentially. The overall incidence of bilateral disease is variable, but the risk of occurrence is higher when there is underlying endocrinopathy, renal disease and obesity. MATERIALS AND METHODS: Amongst the 36 cases (44 hips) of slipped capital femoral epiphysis that we had, there were 8 cases of bilateral disease, of which 3 were bilateral sequential slips. A retrospective review of all case records and X-rays was done. RESULTS: According to Aadalen's criteria, the results were excellent or good in 37 hips, fair in 3 hips, and poor in 2 hips. Two patients were considered as treatment failure because of avascular necrosis. The incidence of bilateral slipped capital femoral epiphysis was 22.2%. The overall average age was 11.4 years. All cases of sequential involvement presented within 15 months of the initial slip. One patient had hypothyroidism (who also had a positive family history for slipped capital femoral epiphysis), while 91.6% were above the 75th percentile for weight. There was no statistically significant difference in age, sex, race, body mass index (BMI) and weight percentile distribution between patients with unilateral and bilateral disease. CONCLUSION: Satisfactory outcomes can be expected in most patients treated for slipped capital femoral epiphysis. The risk of avascular necrosis is higher in unstable slips. The role of prophylactic contra-lateral pinning should be restricted to patients with hypothalamic pituitary-ovarian axis disease, especially hypothyroidism. PMID- 18392296 TI - The relation between insulin resistance determined by homeostatic [corrected from haemostatic] modelling and slow coronary flow. AB - INTRODUCTION: In this study, we sought to determine whether insulin resistance, which is investigated by homeostatic modelling, is related to slow coronary flow (SCF). MATERIALS AND METHODS: A total of 24 patients with SCF (4 females/20 males, mean age 47 +/- 12 years) and 32 patients with normal coronary artery (10 females/22 males, mean age 52 +/- 12 years) were included in the study. Baseline glucose, insulin and plasma lipid levels were measured. A standard oral glucose tolerance test (OGTT) was performed and post-challenge insulin levels were also measured. The index of insulin resistance was calculated with the homeostatic modelling [homeostatic model assessment for insulin resistance index (HOMA-IR)]. RESULTS: There were no differences between the 2 groups with regard to age, lipid levels, blood pressure levels, history of smoking, fasting and post-challenge plasma glucose. Baseline insulin levels were augmented in the SCF group (9.64 +/- 5.93 vs 7.04 +/- 3.26, P = 0.041). HOMA-IR levels were not different between the study groups (2.20 +/- 1.44 vs 1.69 +/- 0.86, P = 0.129). Manifest insulin resistance was significantly higher in the CSF group as compared with the control group (25% vs 3%, P = 0.01). CONCLUSION: Manifest insulin resistance is seen more frequently in patients with SCF. PMID- 18392297 TI - Screening tools for bacteraemia in a selected population of febrile children. AB - INTRODUCTION: This is a prospective, observational study. The aims of the study were to determine the rate of bacteraemia in febrile children in Turkey, and to evaluate the usefulness of white blood cell (WBC) count and manual differential counts of peripheral blood smears and a RISK score in predicting bacteraemia among these children. MATERIALS AND METHODS: A total of 377 febrile children aged 3 to 36 months were included in the study. Complete blood cell (CBC) count, manual differential counts and blood cultures were performed in all patients. The main outcome measures used to evaluate the usefulness of the RISK score were sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), odds ratio (OR), posterior probability, areas under receiver operator characteristic curves (AUC) and miss-to-diagnosis ratio (MDR). RESULTS: Among the patients, 4.4% had bacteraemia and the predominant pathogen was Streptococcus pneumoniae. The Yale Observation Scale scores, percentages of neutrophil and bands, band-neutrophil ratio, absolute neutrophil count and absolute band count were found to be statistically significant predictors of bacteraemia. When the RISK score was 2 or higher, sensitivity was 93.8%, false positive ratio 35.8%, PPV 10.6%, NPV 99.5%, OR 26.2 (95% CI, 3.4 to 200.8), MDR 0.066 and posterior probability value 10%. CONCLUSIONS: We conclude that determination of the RISK score will significantly decrease unnecessary blood culture sampling, antibiotherapy and hospitalisation among febrile patients aged 3 to 36 months without an identifiable focus of infection. PMID- 18392298 TI - The toxicity and outcomes of continuous 5-fluorouracil/cisplatin-based chemotherapy followed by chemoradiation in patients with resected high-risk gastric cancer: results of a single institute. AB - INTRODUCTION: The majority of patients with gastric cancer relapse after definitive surgery and 5-year survival after surgery is very poor. The Intergroup 0116 study showed a modest survival benefit for postoperative bolus 5 fluorouracil-based chemoradiation with a high rate of toxicity. We hypothesised that treatment outcome could be further improved with feasible toxicity using a combination of bolus 5-fluorouracil, continuous 5-fluorouracil, and cisplatin followed by chemoradiation after 3 months of chemotherapy. MATERIALS AND METHODS: Thirty-six patients with stages Ib through IV adenocarcinoma of the stomach or gastrooesophageal junction who had undergone gastric resection and negative margins were assigned to postoperative chemoradiation. The treatment consisted of 6 cycles of continuous 5-fluorouracil (600 mg/m2) for 24 hours, push 5 fluorouracil (400 mg/m2) and leucoverin (LCV) (200 mg/m2) on day 1 to 2 every 2 weeks, cisplatin (60 mg/m2) every 4 weeks followed by combined modality therapy using 45 Gy at 1.8 Gy per day concomitant with weekly bolus 5-fluorouracil (600 mg/m2) and LCV (50 mg). RESULTS: The median age was 59 years (range, 29 to 75) and 25 patients were male. Thirty-five per cent had proximal tumour, T3 or T4 were diagnosed in 92% of the patients and lymph nodes metastases were confirmed in 83%. Grade 3 or 4 neutropaenia was documented in 25%, and gastrointestinal toxicity in 16%. There was no toxic death, but 1 patient had long-term complications. The median disease-free survival was 37.4 months and the overall survival was 40.3 months. CONCLUSIONS: Postoperative chemoradiation with combination of bolus 5-fluorouracil, continuous 5-fluorouracil and cisplatin is a feasible and well-tolerated approach. Larger clinical trials should be conducted to further evaluate the toxicity and the efficacy of this regimen. PMID- 18392299 TI - Seeing the wood for the trees: approaches to teaching and assessing clinical pharmacology and therapeutics in a problem-based learning course. AB - For about 50 years, clinical pharmacology and therapeutics have been taught in the medical schools via traditional lectures and practical classes. During this time, significant changes have occurred in our understanding of medicine and basic sciences. Also the needs for our community have changed dramatically. The explosion of scientific discoveries, the use of new technologies in disease diagnosis, the availability of a wide range of therapeutic options, and the availability of knowledge to everyone via the Internet have necessitated new approaches for teaching medical and other health professional students. Finding information related to a topic has not become a priority in teaching, what has become more important is to teach undergraduate students how to think in addition to what to think. Applying information learnt and assessing its significance in real life situations has become mandatory. The aims of this paper were: (i) to discuss the model we used in introducing clinical pharmacology and therapeutics teaching in the undergraduate course at the University of Melbourne and the educational principles behind the model, and (ii) to discuss the new tools of assessment used in a problem-based learning (PBL) curriculum. PMID- 18392300 TI - Current understanding of the treatment and outcome of acute primary angle-closure glaucoma: an Asian perspective. AB - INTRODUCTION: Primary angle-closure glaucoma (PACG) is a major cause of blindness among Asians. A better understanding of the disease will improve the treatment and outcome of this condition. METHODS: A literature review of all recent publications on PACG was carried out. Articles were retrieved using a key word search of MEDLINE, PubMed and Science Citation Index databases. RESULTS: Following laser peripheral iritodomy for acute angle-closure, Asians were found to have a higher tendency to develop a subsequent rise in intraocular pressure compared to Caucasians. Furthermore, the extent and severity of visual field damage was more severe in Asians than Caucasians, particularly in eyes that presented insidiously with chronic PACG. Prophylactic laser iridotomy in the contralateral eye was found to be highly effective in preventing acute angle closure attacks. CONCLUSION: PACG is more difficult to manage and is associated with more severe long-term visual morbidity in Asians than Caucasians. Regular follow-up of patients with PACG is important for the early detection of progression of the disease and visual field deterioration. PMID- 18392301 TI - Care of women in menopause: sexual function, dysfunction and therapeutic modalities. AB - INTRODUCTION: The physiological changes that occur in menopause alter sexual function and affect well-being. Hormonal changes contribute significantly to reduced sexual function in older women and sexual dysfunction may well be amenable to treatment with exogenous hormones or other agents. MATERIALS AND METHODS: Relevant clinical studies were identified by a computerised literature search. The collated data were presented to fellow gynaecologists for review, analysis of results and discussion in a series of meetings dedicated to finding the best evidence in menopause management. The evidence was assessed and used to prepare guidelines around the management of women who are affected by sexual dysfunction in menopause. RESULTS: Hormone therapy benefits many women who have dyspareunia related to vaginal atrophy, reduced libido and decreased satisfaction, particularly if these symptoms adversely affect their quality of life. Alternative agents such as tibolone and sildenafil citrate can be useful adjuncts. CONCLUSIONS: It is increasingly important to recognise postmenopausal sexual dysfunction. Treatment of this syndrome must be individualised to the specific complaints of each woman. Hormones and other agents are relevant treatment options for properly-selected women. PMID- 18392302 TI - Angioplasty in critical limb ischaemia: one-year limb salvage results. AB - INTRODUCTION: Lower extremity amputation prevention (LEAP) is an ongoing programme in our institution aimed at limb salvage in patients with critical limb ischaemia (CLI). Patients in the LEAP programme with reconstructible anatomy on initial Doppler imaging received either bypass surgery or percutaneous transluminal balloon angioplasty (PTA). We describe the 1-year limb salvage rates in 46 consecutive patients with CLI who received PTA in 2005. CLINICAL PICTURE: A total of 46 patients, 28 women and 18 men, between the ages of 40 and 91 years old (mean age, 70.8) received PTA in 2005. The most common presenting symptom was rest pain (n = 23), followed by pre-existing gangrene (n = 20), non-healing ulcer (n = 17) and cellulitis (n = 8). The majority of the patients (57%) had 3 to 4 risk factors. Diabetes mellitus (91%) and hypertension (80%) were the 2 most common risk factors. The patients were kept under surveillance for periods ranging from 12 to 21 months with a mean of 13.3 months, both clinically and with haemodynamic measurements [ankle-brachial index (ABI), toe pressure (TP) and digital-brachial index (DBI)]. TREATMENT: The aim of PTA is to achieve straight line flow from the abdominal aorta down to either a patent dorsalis pedis or plantar arch with limb salvage as the ultimate goal. The patterns of the treated segments were as follows: aorto-iliac occlusions (n = 3), pure infrapopliteal disease (n = 5), femoropopliteal disease with at least 1 good infrapopliteal run off vessel (n = 16) and combined femoropopliteal and infrapopliteal disease (n = 25). Technical success was achieved in 89% of patients (41 out of 46 patients). The most common cause of technical failure is the inability to cross long chronic total occlusions. OUTCOME: Paired T test was performed and showed statistically significant improvement in haemodynamic markers within the technically successful group. This included increase in the mean ABI from 0.62 (preangioplasty) to 0.91 (Day 1 post-angioplasty), an increase of 0.29 [95% confidence interval (95% CI), 0.1953 to 0.3875; P <0.001]. One year post-angioplasty, the mean ABI was 0.84, an increase of 0.22 (95% CI 0.1512 to 0.3121; P <0.001). There was also significant increase in the mean DBI of 0.17 from 0.23 to 0.41 (pre-angioplasty versus Day 1 post angioplasty - 95% CI of 0.1006 to 0.2433; P <0.001). In addition, significant increase in the mean TP of 28.2 mmHg from 36.8 to 63.2 mmHg (pre angioplasty versus Day 1 post angioplasty - 95% CI, 18.493 to 37.939; P <0.001) was also noted. Of the 23 patients who presented with rest pain, total abolishment of symptoms was achieved in 21 patients (91%). Healing of pre existing gangrene was attained in 15 patients (66%). Five patients subsequently received minor amputation for pre-existing gangrene. Clinical improvement in all the patients who presented with non-healing ulcers (n = 17) and cellulitis (n = 8) was attained. More importantly, all healed ulcers remained healed throughout the study period. The limb salvage rates were 93% at 1 month, 87% at 3 months, 82% at 6 months and 78% at 1 year. CONCLUSION: Angioplasty is a safe and effective limb salvage method in patients with CLI and has a high 1-year limb salvage rate. PMID- 18392303 TI - Treatment of cardiogenic pulmonary oedema by helmet-delivered non-invasive pressure support ventilation in children with scorpion sting envenomation. AB - INTRODUCTION: The aim of this study was to evaluate the feasibility of non invasive positive pressure ventilation through a new interface helmet in the treatment of cardiogenic pulmonary oedema due to scorpion sting envenomation in children. CLINICAL PICTURE: Three patients presented with fever, and respiratory distress following scorpion sting. Their cardiac enzymes were abnormal. Electrocardiogram (ECG) of 3 patients showed features of myocardial strain with ST elevation. Bedside chest X-ray taken in emergency showed marked bilateral infiltrates suggestive of pulmonary oedema. M-mode, two-dimensional colour-flow Doppler echocardiogram showed left ventricular dysfunction. TREATMENT AND OUTCOME: At paediatric intensive care unit admission, they were treated with antivenom, prazosin (0.03 mg/kg/dose), dopamine (15 mcg/kg/ min), dobutamine (10 mcg/kg/min) and nitroprussid (1 mcg/kg/min). Epinephrine (0.1 mcg/kg/ min) were added later. They were hypoxic and dyspnoeic. A slight sedation was induced with ketamine and/or midazolam. Non-invasive pressure support ventilation (NPSV) was delivered via the helmet by means of an intensive care unit ventilator. We evaluated the effect of NPSV delivered by helmet on oxygenation, respiratory rate, haemodynamics, complications and outcome. An improvement of oxygenation was observed within 2 hours of treatment.The helmet was well tolerated by all the children. No complications occurred in the 3 patients. CONCLUSION: This new approach of delivering NPSV through a helmet allows the successful treatment of cardiogenic pulmonary oedema in children with scorpion sting envenomation, assuring a good tolerance without complications. Future studies are needed before recommending the extensive application of this technique in all cases of cardiogenic pulmonary oedema due to scorpion sting envenomation. PMID- 18392304 TI - Pathological pulmonary hernia in a patient with metastatic breast cancer. AB - INTRODUCTION: Pathological pulmonary hernia is a rare clinical entity which can be caused by malignancies. CLINICAL PICTURE: A 72-year-old female presented with a painful bulge in the left 4th intercostal space. Chest radiography and computed tomography demonstrated a left pulmonary hernia, pleural effusion and destruction of ribs. TREATMENT: The hernia sac was excised and a part of the chest wall was resected with reconstruction of residual defect. OUTCOME: The patient died 2 years after the treatment. CONCLUSIONS: A multidisciplinary approach involving various medical specialists may offer patients with pathological pulmonary hernia remarkable palliation and better quality of life. PMID- 18392305 TI - Fitness to work: legal pitfalls. AB - Medical examinations for fitness to work are undertaken by the specialist and the nonspecialist occupational physicians alike. An assessment done negligently in such a capacity will expose physicians to legal risks, even if they are not specialist occupational physicians. This is not unlike negligent care given in the traditional therapeutic care setting. Much of the case law governing legal disputes in medical fitness to work assessments depend on the kind of loss that is at stake, that is, whether it is one resulting in economic loss, as in loss of an employment opportunity, or one resulting in personal injury, such as when the doctor negligently fails to diagnose or to communicate a serious medical finding. In the latter, the courts are more likely to find for the injured claimants. One of the ways for doctors to reduce their risk in this area is to establish with prospective employers and employees the ground rules when conducting medical fitness to work examinations. PMID- 18392306 TI - Successful surgical treatment of renal cell carcinoma with calvarial metastases. PMID- 18392307 TI - A case of myopericarditis in a patient with methicillin-resistant Staphylococcus aureus community-acquired pneumonia. PMID- 18392308 TI - Non-compaction cardiomyopathy presenting with classical angina pectoris. PMID- 18392309 TI - Renal cell carcinoma bony metastasis treatment. PMID- 18392310 TI - Advanced Burkitt's lymphoma presenting with jejunal perforation. PMID- 18392311 TI - Erysipelothrix rhuseopathiae septicaemia with prolonged hypotension: a case report. PMID- 18392312 TI - Pericardial thymoma: an unusual cause of sudden death. PMID- 18392313 TI - The Tao of bao: a randomised controlled trial examining the effect of steamed bun consumption on night-call inpatient course and mortality. AB - BACKGROUND: Medical superstitions remain prevalent in today's stressful and technology driven healthcare environment. These irrational beliefs commonly involve night calls, which are periods of volatile workload. In Singapore and Hong Kong, it is commonly held that consumption of steamed buns ("bao") by on call physicians is associated with increased patient admissions and mortality, due to a homonymous interpretation of the word "bao" in dialect. MATERIALS AND METHODS: A prospective unblinded randomised controlled trial with a permuted block randomisation design was performed on weekdays over 6 weeks. Steamed buns or control food were offered to the internal medicine night-call team of a tertiary-care hospital on a nightly basis. Information on admissions and mortality was collected from the hospital electronic database. Data on sleep patterns and shift duration were obtained by interview. RESULTS: There were no significant differences in the median number of hours slept on days on "bao" administration versus "control" intervention (2 +/- median absolute variation of 1.5 h vs 2 +/- 1.5 h, P = 0.30) or in the number of hours spent in the hospital (30.8 +/- 1.9 h vs 30.5 +/- 2.2 h, P = 0.09). There were no significant differences in the median number of general ward admissions per night (n = 73 +/- 6 versus 71 +/- 7 admissions, P = 0.35), monitored care unit admissions (4 +/- 1.5 vs 4 +/- 1.5 admissions, P = 0.65) or inpatient mortality (2 +/- 1.5 vs 2 +/- 1.5 deaths per night, P = 0.47). CONCLUSION: The consumption of steamed buns ("bao") has no effect on inpatient admissions, mortality, or sleep duration on call. Regardless, our results indicate that the night call in Singapore remains a challenge in terms of workload and shift duration. PMID- 18392314 TI - [46th Congress of the German Society of Nuclear Medicine. Introduction]. PMID- 18392315 TI - [Routine measurement of serum calcitonin in patients with nodular thyroid disorders?]. AB - In spite of the fact that the German Society of Endocrinology has recommended calcitonin as screening-parameter the majority of physicians in Germany do not routinely use calcitonin in patients with thyroid nodules to exclude medullary thyroid cancer (MTC). The future revision of the recommendation should describe reference values for each commercially available assay, separately for men and women (basal and after pentagastrin-stimulation), and should define sonomorphological inclusion criteria. The epidemiological database of the prevalence of MTC is controversial and the specificity of basal elevated calcitonin levels is limited up to the 5-fold of the upper reference level. If renal insufficiency, bacterial infection, and an alcohol- or drug-induced stimulation of calcitonin is excluded, hypercalcitoninaemia should be confirmed by a second measurement (if necessary using another assay). Stimulation of calcitonin by use of pentagastrin is mandatory prior to the decision on thyroidectomy. A stimulated calcitonin level < 100 pg/ml justifies "wait and see". If stimulated calcitonin levels range between 100 and 200 pg/ml or higher, the differentiation between C-cell hyperplasia and MTC remains uncertain, especially in men. The implementation of calcitonin-screening requires the definition of sonographic inclusion criteria and validation of each assay. Additional pre-requisites are excellent logistic (short period between blood sampling and start of the laboratory test), knowledge of differential diagnoses, knowledge of the consumption of drugs and alcohol, availability of pentagastrin testing and of moleculargenetic testing with full information to the patients and sufficient time before the decision on surgery is made. All this and the choice of a skilled surgeon, experienced in thyroidectomy and lymphadenectomy with a low rate of local complications are the rationale to recommend calcitonin-screening primarily in centers for thyroid disorders. PMID- 18392316 TI - Multi-modality imaging of uveal melanomas using combined PET/CT, high-resolution PET and MR imaging. AB - We investigated the efficacy of combined FDG-PET/CT imaging for the diagnosis of small-size uveal melanomas and the feasibility of combining separate, high resolution (HR) FDG-PET with MRI for its improved localization and detection. PATIENTS, METHODS: 3 patients with small-size uveal melanomas (0.2-1.5 ml) were imaged on a combined whole-body PET/CT, a HR brain-PET, and a 1.5 T MRI. Static, contrast-enhanced FDG-PET/CT imaging was performed of head and torso with CT contrast enhancement. HR PET imaging was performed in dynamic mode 0-180 min post injection of FDG. MRI imaging was performed using a high-resolution small-loop coil placed over the eye in question with T2-3D-TSE and T1-3D-SE with 18 ml Gd contrast. Patients had their eyes shaded during the scans. Lesion visibility on high-resolution FDG-PET images was graded for confidence: 1: none, 2: suggestive, 3: clear. Mean tumour activity was calculated for summed image frames that resulted in confidence grades 2 and 3. Whole-body FDG-PET/CT images were reviewed for lesions. PET-MRI and PET/CT-MRI images of the head were co-registered for potentially improved lesion delineation. RESULTS: Whole-body FDG-PET/CT images of 3/3 patients were positive for uveal melanomas and negative for disseminated disease. HR FDG-PET was positive already in the early time frames. One patient exhibited rising tumour activity with increasing uptake time on FDG-PET. MRI images of the eye were co-registered successfully to FDG-PET/CT using a manual alignment approach. CONCLUSIONS: Small-size uveal melanomas can be detected with whole-body FDG-PET/CT. This feasibility study suggests the exploration of HR FDG PET in order to provide additional diagnostic information on patients with uveal melanomas. First results support extended uptake times and high-sensitivity PET for improved tumour visibility. MRI/PET co-registration is feasible and provides correlated functional and anatomical information that may support alternative therapy regimens. PMID- 18392317 TI - Uptake of (18)F-FLT and (18)F-FDG in primary head and neck cancer correlates with survival. AB - The aim of the study was to determine the practicability of (18)F-FLT in tumours of the head and neck area in terms of visualization, a possible correlation between FLT uptake and proliferation fraction as determined by Ki-67 immunostaining, and if tumoural FLT-uptake has a prognostic meaning, as determined by a correlation to patient survival time. Results were compared to (18)F-FDG. PATIENTS, METHODS: 20 patients with previously untreated lesions of the head and neck area, which were clinically highly suspicious to be malignant, underwent PET scans with (18)F-FLT and (18)F-FDG, a CT of the head and neck area, and a biopsy. Tumour tracer uptake was determined by standardized uptake value (SUV) normalized to body weight and /non-tumor ratios (T/N). (18)F-FDG and (18)F FLT uptake were compared with histopathologic and immunohistochemical results. RESULTS: 19 patients had malignant tumours; one patient had a benign cystadenoma (so called Warthin's tumour) of the parotid gland. One negative lesion turned out to be a malignant T1 stage squamous cell carcinoma in both PET scans, the Warthin's tumour was false positive with (18)F-FDG but showed only faint uptake with (18)F-FLT, resulting in a sensitivity of 95 % for both tracers. Of all lesions, maximum SUVs of (18)F-FLT ranged from 1.53 to 11.70 (mean +/- SD 5.81 +/ 2.28) those of FDG from 2.63 to 16.50 (mean +/- SD 8.91 +/- 3.58), p < 0.001. (18)F-FLT-T/N ranged from 0.94 to 5.85 (mean +/- SD, 3.18 +/- 1.21), (18)F-FDG T/N was from 0.92 to 7.50 (mean +/- SD, 3.6 +/- 1.74), n.s. The mean survival time was 18 months in a maximum follow up time of 36 months. A significant correlation between both PET tracers and survival was detected, but no correlation between the amount of Ki-67 positive cells and FLT. CONCLUSION: In head and neck cancer in the primary setting (18)F-FLT does not provide additional visual information in comparison to (18)F-FDG.(18)F-FLT uptake is inversely correlated with patient survival, as well as (18)F-FDG. PMID- 18392318 TI - Angiogenesis and the progress of vascular and tumor biology: a tribute to Judah Folkman. PMID- 18392319 TI - Expanding the role of coagulation in arterial thrombosis: evidence from animal models using a new factor Xa inhibitor. PMID- 18392320 TI - Factor VIIa gets even bigger. PMID- 18392321 TI - Recurrent venous thromboembolism: quite harmless after all? PMID- 18392322 TI - Cell-derived microparticles: a mediator of inflammation in aortic valve stenosis? PMID- 18392323 TI - Prolonged in-vivo half-life of factor VIIa by fusion to albumin. AB - For the treatment of haemophilia patients with inhibitors, recombinant factor VIIa (rFVIIa) is available as a therapeutic option to control bleeding episodes with a good balance of safety and efficacy. However, the short in-vivo half-life of approximately 2.5 hours makes multiple injections necessary, which is inconvenient for both physicians and patients. Here we describe the generation of a recombinant FVIIa molecule with an extended half-life based on genetic fusion to human albumin. The recombinant FVII albumin fusion protein (rVII-FP) was expressed in mammalian cells and upon activation displayed a FVII activity close to that of wild type FVIIa. Pharmacokinetic studies in rats demonstrated that the half-life of the activated recombinant FVII albumin fusion protein (rVIIa-FP) was extended six- to seven-fold compared with wild type rFVIIa. The in-vitro and in vivo efficacy was evaluated and was found to be comparable to a commercially available rFVIIa (NovoSeven((R))). The results of this study demonstrate that it is feasible to develop a half-life extended FVIIa molecule with haemostatic properties very similar to the wild-type factor. PMID- 18392324 TI - Cleavage of factor XIII by human neutrophil elastase results in a novel active truncated form of factor XIII A subunit. AB - The first step in the activation of plasma factor XIII (FXIII) is the cleavage of R37-G38 bond in FXIII-A subunit (FXIII-A) by thrombin, which makes the subsequent formation of an active transglutaminase possible. No active truncated form of FXIII-A, other than G38-FXIII-A, has been identified. In contrast to thrombin, which has a preference toward arginine residues, human neutrophil elastase (HNE) cleaves peptide bonds at small side-chain aliphatic amino acids, preferably at valine. As there are several valine residues close to the thrombin cleavage-site, we tested if an active truncated FXIII-A was formed during fragmentation of FXIII by HNE. It was demonstrated by Western blotting and transglutaminase assay that HNE induced a limited cleavage of FXIII-A resulting in the activation of both plasma and cellular FXIII; the maximal transglutaminase activities were 52.5% and 67.4% of thrombin-activated FXIII, respectively. After the relatively rapid activation a much slower inactivation occurred. HNE-activated FXIII cross-linked fibrin gamma- and alpha-chains in the clot formed by batroxobin moojeni. MALDI TOF analysis of the cleaved fragments and N-terminal Edman degradation of the truncated protein identified V39-N40 as the primary cleavage-site and N40-FXIII-A as the active form. No primary cleavage occurred at V34, V35, V47, V50 residues. FXIII-A V34L polymorphism, which increases the rate of FXIII-A cleavage by thrombin, was without effect on FXIII activation by HNE. Molecular modeling located the primary HNE cleavage-site in the middle of the flexible and accessible Q32-L45 loop and showed that other neighboring valine residues were in less favorable position. PMID- 18392325 TI - Inherited thrombophilic abnormalities and risk of portal vein thrombosis. a meta analysis. AB - Inherited thrombophilic abnormalities may have a role in the development of portal vein thrombosis (PVT). However, the prevalence of these factors in patients with PVT has been evaluated only in small studies with non-conclusive results. It was the purpose of this study to assess the risk of PVT associated with factor V Leiden (FVL) and G20210A prothrombin mutation (PTM). The MEDLINE, EMBASE, Cochrane Library databases, reference lists of retrieved articles and contact with content experts were used. Studies carried out in Western Europe comparing the prevalence of prothrombotic abnormalities in patients with PVT and in controls without a history of thromboembolic disease were included. Two reviewers independently selected studies and extracted study characteristics, quality and outcomes. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for each trial and pooled using a fixed and random-effects model. Statistical heterogeneity was evaluated using the I(2) statistic. Sensitivity analyses were performed examining separately studies according to the etiology of PVT and to control population. Twelve studies involving more than 3,000 patients were included. The pooled OR for PVT was 1.90 (95%CI: 1.25, 2.90) in patients with FVL and 4.48 (95%CI: 3.10, 6.48) in patients with PTM. In conclusion, PVT is associated with the presence of FVL and PTM in Western Europe. PMID- 18392326 TI - Death due to recurrent thromboembolism among younger healthier individuals hospitalized for idiopathic pulmonary embolism. AB - The incidence of death due to recurrent pulmonary embolism (PE) after a first time idiopathic PE is not well defined. We conducted a retrospective study of patients age 18 to 56 years who had idiopathic PE between 1994-2001. The incidence and cause of death within five years was determined using linked discharge records and a master death registry. A total of 3,456 patients had a first-time idiopathic PE. The rate of recurrent VTE 0-6 months after the index event was 13.1%/year, and 2.9%/year 6-60 months after the event. During the mean follow-up of 3.2 years 118 (3.4%, 95% confidence interval [CI] = 2.8-4.1%) patients died. Fifty-two (44%) deaths occurred <29 days after the index PE (case fatality rate = 1.5%, 95%CI = 1.1-2.0%). Among the 66 cases (1.9%) that died after 28 days, 18 (0.52%) were due to recurrent PE or its sequelae: eight had recurrent PE alone, five had recurrent PE and a serious co-morbid illness, and five had thromboembolic pulmonary hypertension with or without acute PE. The person-time rate of death (deaths per 100 patient-years) attributed to any recurrent thromboembolism 6-60 months after the event was 0.16% (95%CI = 0.1 0.26%). Ten of the 18 (56%) late thromboembolic deaths reflected a first-time recurrent PE. The 28-day case-fatality rate for recurrent VTE was 2.8% (95%CI = 1.5-4.9%). In this cohort of younger patients with idiopathic PE, the rate of death due to recurrent VTE, particularly to first-time recurrent PE, was low. Among the patients who died of thromboembolism >28 days after the index PE, 28% had developed pulmonary hypertension. PMID- 18392327 TI - Glycaemic control improves fibrin network characteristics in type 2 diabetes - a purified fibrinogen model. AB - Diabetic subjects have been shown to have altered fibrin network structures. One proposed mechanism for this is non-enzymatic glycation of fibrinogen due to high blood glucose. We investigated whether glycaemic control would result in altered fibrin network structures due to decreased fibrinogen glycation. Twenty uncontrolled type 2 diabetic subjects were treated with insulin in order to achieve glycaemic control. Twenty age- and body mass index (BMI)-matched non diabetic subjects were included as a reference group. Purified fibrinogen, isolated from plasma samples was used for analysis. There was a significant decrease in fibrinogen glycation (6.81 to 5.02 mol glucose/mol fibrinogen) with a corresponding decrease in rate of lateral aggregation (5.86 to 4.62) and increased permeability (2.45 to 2.85 x 10(-8) cm(2)) and lysis rate (3.08 to 3.27 microm/min) in the diabetic subjects after glycaemic control. These variables correlated with markers of glycaemic control. Fibrin clots of non-diabetic subjects had a significantly higher ratio of inelastic to elastic deformation than the diabetic subjects (0.10 vs. 0.09). Although there was no difference in median fiber diameter between diabetic and non-diabetic subjects, there was a small increase in the proportion of thicker fibers in the diabetic samples after glycaemic control. Results from SDS-PAGE indicated no detectable difference in factor XIIIa-crosslinking of fibrin clots between uncontrolled and controlled diabetic samples. Diabetic subjects may have altered fibrin network formation kinetics which contributes to decreased pore size and lysis rate of fibrin clots. Achievement of glycaemic control and decreased fibrinogen glycation level improves permeability and lysis rates in a purified fibrinogen model. PMID- 18392328 TI - Allosteric regulation of alpha(IIb)beta(3) by beta(3) 95-105. AB - The linear recognition sequences of an anti-beta(3) antibody that blocked platelet aggregation were identified using beta(3) tryptic peptides. Two of these recognition sequence-containing peptides were mapped to beta(3) 92-105, and antibodies affinity purified using these peptides blocked platelet aggregation. Examining the structure of alpha(IIb)beta(3) identified beta(3) 95-105 as the solvent accessible sequence within beta(3) 92-105. A peptide corresponding to beta(3) 95-105 was synthesized and used to affinity purify the beta(3) antibody. Anti-beta(3) 95-105 completely blocked platelet aggregation and agonist-induced fibrinogen binding to platelets, but had no effect on cyclic-RGD binding. Binding of anti-beta(3) 95-105 to alpha(IIb)beta(3) also did not alter the structure of the alpha(IIb) cap subdomain, as measured by anti-alpha(IIb) 201-217 binding. beta(3) 95-105 and peptides spanning two adjacent sequences in the structure of beta(3) did not bind fibrinogen and were ineffectual in blocking agonist-induced platelet aggregation. Structure analysis revealed that beta(3) 95-105 is adjacent to one of the two hinges in beta(3) that allows for the outward swing of the hybrid and PSI domains which is central to the conversion of alpha(IIb)beta(3) from a low into a high affinity state. Thus, the binding of an antibody to beta(3) 95-105 could serve as a fulcrum for allosteric regulation of alpha(IIb)beta(3) by regulating the movement of the hybrid-PSI domain. PMID- 18392329 TI - Increased levels of circulating microparticles in patients with severe aortic valve stenosis. AB - The mechanisms of the progression of aortic valve stenosis are unknown. The involvement of mononuclear cells and of chronic systemic inflammation has been suggested by analysis of pathological specimens. We hypothesize that shear stress caused by the constricted aortic orifice contributes to systemic proinflammation by activation of circulating blood cells and thereby generation of microparticles. Using flow cytometry we analyzed 22 patients with severe aortic valve stenosis (AVS) and 18 patient controls for the generation of circulating microparticles from platelet- (PMPs: CD31(+)/CD61(+) or CD62P(+)), leukocyte- (LMPs: CD11b(+)) and endothelial cell (EMPs: CD62E(+)) origin. Apart from the constricted valve orifice groups were similar. PMPs were increased in AVS patients and their number correlated with valvular shear stress. Monocytes were activated in AVS patients, an observation that was also reflected by increased numbers of LMPs and by the detection of PMP-monocyte conjugates. Furthermore, EMPs reflecting the activation of endothelial cells but also conferring systemic inflammatory activity were increased in AVS patients and correlated with the number of activated monocytes. In conclusion, we show that AVS is accompanied by increased levels of microparticles and that shear stress can induce the formation of microparticles. Based on our results and histologic findings of other investigators the speculation that shear stress related to aortic valve stenosis induces a vicious circle including the generation of PMPs, the subsequent activation of monocytes and LMPs and finally the activation of endothelial cells contributing to the progress of aortic valve stenosis appears to be justified. PMID- 18392330 TI - High-dose erythropoietin alters platelet reactivity and bleeding time in rodents in contrast to the neuroprotective variant carbamyl-erythropoietin (CEPO). AB - The haematopoietic hormone erythropoietin (EPO) has neuroprotective properties and is currently being explored for treatment of stroke and other neurological disorders. Short-term, high-dose treatment with EPO seems to improve neurological function of stroke patients but may be associated with increased thrombotic risk, whereas alternative non-erythropoietic neuroprotective derivatives of EPO, such as carbamylated EPO (CEPO), may be devoid of such side-effects. We investigated the effects of short-term, high-dose treatment with EPO and CEPO on platelet function and haemostasis in healthy mice and rats. Animals received three daily doses of EPO or CEPO (50 microg/kg), and blood was compared with respect to alterations in haematology and platelet reactivity. In rats, treatment with EPO increased the haematocrit to >50% and the mean platelet volume by 37%, while CEPO had no effect on these parameters. Platelets from EPO-treated rats showed an increased sensitivity to thrombin receptor agonist peptides and elevated plasma levels of soluble P-selectin (sP-selectin) were found in treated mice. Further indicators of platelet hyperreactivity in EPO, but not CEPO-treated animals, were significantly increased aggregatory responses to collagen in whole blood and platelet-rich plasma (PRP). The increased platelet reactivity was paralleled by a decreased bleeding time after tail transection in rats. Samples from EPO-treated rats showed an attenuated response to ADP in whole blood aggregometry and thrombelastography (TEG) platelet mapping but not in apyrase-treated PRP, suggesting involvement of ADP receptor desensitization. These findings suggest that while EPO affects various aspects of platelet function, CEPO is devoid of such effects. PMID- 18392331 TI - Mutation of human plasminogen kringle 1-5 enhances anti-angiogenic action via increased interaction with integrin alpha(v)beta(3). AB - Angiogenesis plays a primary role in tumor growth and metastasis. Angiostatin, a proteolytic fragment containing the first four kringle domains of human plasminogen, can inhibit angiogenesis. The anti-angiogenic activities of kringle 1-5 (K(1-5)) and kringle 5 fragments of plasminogen are greater than angiostatin in inhibiting angiogenesis and angiogenesis-dependent tumor growth. To further optimize kringle fragment anti-angiogenic activities, mutations were created at the potential glycosylation sites Asn-289 and Thr-346 and the Lys binding site, Leu-532, at kringle 5, including K(1-5)N289A (replacing Asn by Ala at residue 289), K(1-5)T346A, K(1-5)L532R, K(1-5)N289A/T346A, K(1-5)T346A/L532R, K(1 5)N289A/L532R, and K(1-5)N289A/T346A/L532R. Wild-type and mutant K(1-5) proteins were expressed successfully by the Pichia pastoris expression system. Native K(1 5) from proteolytic cleavage and wild-type K(1-5) have similar activity in inhibiting basic fibroblast growth factor-induced endothelial cell proliferation. Among these mutated proteins, K(1-5)N289A/T346A/L532R exhibited the greatest effect in inhibiting endothelial cell proliferation and in inducing endothelial cell apoptosis. Integrin alpha(v)beta(3)-mediated adhesion of K(1 5)N289A/T346A/L532R to endothelial cells was more greatly enhanced when compared to wild type K(1-5). Furthermore, K(1-5)N289A/T346A/L532R was most potent in inhibiting basic fibroblast growth factor-induced angiogenesis in Matrigel assay in vivo. Angiogenesis-dependent tumor growth was inhibited by systemically injected K(1-5)N289A/T346A/L532R into mice. These results demonstrate that alteration of glycosylation and Lys binding properties could increase the anti angiogenic action of K(1-5), possibly via enhanced interaction with integrin alpha(v)beta(3) in endothelial cells. PMID- 18392332 TI - Time course of prothrombotic and proinflammatory substance release after intracoronary stent implantation. AB - We hypothesized that restenosis after coronary stenting is predicted by elevated levels of markers of thrombus formation and inflammation. Plasma levels of representative markers of inflammation, the thrombin and plasmin activation systems and adhesion molecules were measured in 59 patients with stable angina pectoris before, immediately after and 6 hours (h), 12 h, 24 h, one month and six months after elective stent implantation (radioactive phosphorus-32 stents/RSs/ n = 16, bare-metal stents/BMSs/ n = 43). All patients underwent clinical and angiographic follow-up (FUP) six months after stenting. RSs had significantly higher angiographic severity of restenosis than BMSs (47.1 +/- 20.1% vs. 27.6 +/- 22.0%, p = 0.003). Repeated measures ANOVA revealed significant differences between the BMS and RS groups as regards the increases in plasma levels of vascular cell adhesion molecule-1 (VCAM-1, p = 0.022), plasminogen activator inhibitor-1 (PAI-1, p = 0.047), tissue-type plasminogen activator (tPA, p = 0.047) and CD40 ligand (CD40L, p = 0.038). tPA levels tended to increase immediately after stenting in both groups, whereas the PAI-1 level one month after stenting was elevated significantly only in the RS group. In the RS group, the plasma levels of CD40L were increased at 24 h and six months after stenting, and the VCAM-1 level rose immediately after stenting and remained high during the FUP. Multivariate analysis on pooled laboratory data of both groups revealed elevated levels of VCAM-1 at 12 h and at six months as significant predictors of the severity of stent restenosis. In conclusion, the process of inflammation and thrombosis occurring after coronary interventions seems to be prolonged and enhanced in patients with high-grade restenosis at the follow up. PMID- 18392333 TI - Dose-dependent thrombus resolution due to oral plaminogen activator inhibitor (PAI)-1 inhibition with tiplaxtinin in a rat stenosis model of venous thrombosis. AB - This study aimed to evaluate a small-molecule PAI-1 inhibitor (PAI-039; tiplaxtinin) in a rodent stenosis model of venous thrombosis in a two-phase experiment. Phase 1 determined the efficacy of tiplaxtinin against Lovenox (LOV), while phase 2 determined the dose-dependent efficacy. For both phases, drug treatment began 24 hours after surgically induced venous thrombosis and continued for four days. Phase 1 animals (n = 24) receiving low-dose (LD; 1 mg/kg oral gavage) PAI-1 inhibitor demonstrated a 52% decrease in thrombus weight (TW) versus controls (p < 0.05) with significant reductions in active plasma PAI-1, while the high-dose (HD; 10 mg/kg oral gavage) group demonstrated a 23% reduction in TW versus controls. Animals treated subcutaneously with LOV (3 mg/kg) showed a 39% decrease in TW versus controls (p < 0.05). Coagulation tests (aPTT and TCT) were significantly different in LOV compared to PAI-1 inhibitor groups. PAI-039 treatment was also associated with significantly increased return of inferior vena cava blood flow four days post-thrombosis versus controls (p < 0.05). In phase 2 (n = 30), TW was reduced from the 0.5 mg/kg to 5 mg/kg experimental groups, with the 10 mg/kg group demonstrating a paradoxical increase. The 5 mg/kg group showed statistically significant decreases in TW versus controls after four treatment days (p < 0.05). This is the first study to demonstrate dose related effects of PAI-039 on increasing thrombus resolution and inferior vena cava blood flow without adverse effects on anti-coagulation in a rat stenosis model of venous thrombosis. PMID- 18392334 TI - Comparison of PD0348292, a selective factor Xa inhibitor, to antiplatelet agents for the inhibition of arterial thrombosis. AB - The objective of this study was to determine if orally-administered PD0348292, a direct specific factor Xa inhibitor, inhibits thrombosis following porcine carotid arterial injury comparably to aspirin or clopidogrel alone or in combination. We further sought to determine whether the antithrombotic efficacy in vivo could be predicted using an ex-vivo perfusion chamber. Oral treatments included: PD0348292 (0.4, 0.9, or 4.3 mg/kg); PD0348292 (0.4 mg/kg) plus aspirin (325 mg); aspirin; clopidogrel (75 mg); aspirin plus clopidogrel; or vehicle (n = 6-10/group). Aspirin and clopidogrel were administered 27 and four hours pre injury and PD0348292 or vehicle was administered four hours pre-injury. Both carotid arteries were crush-injured, and thrombus was measured by detection of (111)In-platelets over 30 minutes. Prior to injury, the antithrombotic efficacy was assessed by ex-vivo perfusion chamber platelet deposition. PD0348292 produced dose-dependent prothrombin time (0.9- to 2.9-fold) and aPTT (1.4- to 2.5-fold) prolongations. Bleeding times were significantly prolonged in each active drug group compared to vehicle, but were not significantly different between drug groups. PD0348292 significantly inhibited arterial platelet deposition (x10(6)/cm(2)) at 4.3(549 +/- 1,066), 0.9 (399 +/- 162) and 0.4 mg/kg (531 +/- 470) compared to vehicle (2,242 +/- 1,443). Aspirin (992 +/- 973), clopidogrel (537 +/- 483), clopidogrel plus aspirin (228 +/- 66) or PD0348292 plus aspirin (558 +/- 317) also significantly inhibited platelet deposition, although these values were not significantly different than with any dose of PD348292. Perfusion chamber platelet deposition correlated significantly with in-vivo anti-thrombotic response. In conclusion, PD0348292 inhibited arterial thrombosis comparable to aspirin plus clopidogrel. Perfusion chamber methodology may be useful in predicting in-vivo antithrombotic efficacy. PMID- 18392335 TI - Influence of coagulation factors and tissue factor concentration on the thrombin generation test in plasma. AB - The thrombin generation test is used to study coagulation in patients with haemorrhagic diseases or with high thrombotic risk. To our knowledge, this is the first study investigating the relative influence of coagulation factors on thrombin generation in plasma. The aim was to investigate the influence of coagulant factors, anticoagulant factors, and tissue factor (TF) on three parameters: endogenous thrombin potential (ETP), peak thrombin concentration, and lag time for the appearance of thrombin. At a low TF concentration, all factors except factor XI influenced thrombin generation. At a high TF concentration, only the factors of the extrinsic pathway exerted an influence. ETP and peak thrombin were linearly correlated to factor II concentration. Factor V and factor VII effects increased hyperbolically with factor concentration. The influence of factor X on thrombin generation depended on TF concentration. In the absence of factor VIII and factor IX, ETP fell to 60-70% of the normal when peak thrombin fell to 25-30% of the normal. Fibrinogen concentration influenced ETP and peak thrombin and decreasing fibrinogen levels shortened the lag time. As expected, decreasing antithrombin concentration caused dramatic increases in thrombin generation. Protein S prolonged the lag time, especially at a low TF concentration. No effect of protein C was observed, likely due to the absence of thrombomodulin. The thrombin generation test was more sensitive to factor deficiencies at low than at high TF concentration. ETP was not the most critical parameter for studying coagulation factor deficiencies. Instead, peak thrombin was the most sensitive parameter. PMID- 18392336 TI - Unique heterozygous intron 22 inversion band pattern in a haemophilic male detected by long polymerase chain reaction (PCR). PMID- 18392337 TI - Alternative anticoagulation with danaparoid in two pregnancies in a patient with former heparin-induced thrombocytopenia (HIT), homozygous factor V Leiden mutation, a history of venous thrombosis and recurrent pregnancy losses. PMID- 18392338 TI - Fondaparinux-related thrombocytopenia in a previous low-molecular-weight heparin (LMWH)-induced heparin-induced thrombocytopenia (HIT). PMID- 18392339 TI - Rituximab failure in a patient with monoclonal gammopathy of undetermined significance (MGUS)-associated acquired von Willebrand syndrome. PMID- 18392340 TI - Identification of 5-HT3 receptors on human platelets: increased surface immunoreactivity after activation with adenosine diphosphate (ADP) and thrombin receptor-activating peptide (TRAP). PMID- 18392341 TI - High factor VIIa levels do not promote tumor metastasis. PMID- 18392342 TI - Influence of low-density lipoprotein (LDL) receptor-related protein and ABO blood group genotypes on factor XI levels. PMID- 18392343 TI - Thrombin regulation in neonates undergoing cardiopulmonary bypass. PMID- 18392344 TI - Risk factors and clinical presentation of venous thromboembolism according to the age of relatives of patients with factor V Leiden. PMID- 18392345 TI - RANTES is required for ischaemia-induced angiogenesis, which may hamper RANTES targeted anti-atherosclerotic therapy. PMID- 18392346 TI - Preventing venous thromboembolism in US hospitals: are surgical patients receiving appropriate prophylaxis? PMID- 18392347 TI - The effects of prenatal and postnatal malnutrition on the morphology, differentiation, and metabolism of skeletal striated muscle tissue in rats. AB - OBJECTIVE: To study the contractile properties, metabolism and morphological characteristics of muscles submitted to prenatal and postnatal protein malnutrition. METHODS: Animals were distributed into two groups: Control, normoprotein diet (CG; n = 15; 5/5/5), and Malnourished, hypoprotein diet (MG; n = 15; 5/5/5), and examined on the 7th, 14th, and 28th days of the experiment. Total body mass, weight, and the contractile properties and morphology of the anterior tibial muscle were assessed. Several 8 microm-thick tissue samples were taken from 7, 14, and 28 day old rats and stained with HE or subjected to NADH-TR or m-ATPase (pH = 4.4) techniques. RESULTS: Body and muscle weight were lower in the malnourished group. On the 7th day of malnutrition, muscle samples exhibited fibers with smaller diameter, higher polymorphism and higher endomysial conjunctive tissue content. Histochemical methods were unable to precisely determine the types of fiber present. On the 14th day, there were smaller muscle fibers, more polymorphism, many of them with central nuclei and moderate endomysial conjunctive tissue content. With reference to contractile properties, the m-ATPase reaction identified both slow and fast fibers. The NADH-TR reaction revealed the following types of fiber: slow oxidative (SO), fast oxidative glycolytic (FOG) and fast glycolytic (FG). On the 28th day smaller, bunched muscle fibers varying shapes. All three types of fiber exhibited unclear recognition limits with respect to contraction and metabolism. CONCLUSION: Our experimental results suggest that, in addition to the reduction in numbers of fibers, malnutrition retards the differentiation of the morphological, metabolic, and contractile characteristics of skeletal muscle fibers in growing rats. PMID- 18392348 TI - The current context for research in Public Oral Health. PMID- 18392349 TI - [Biological and socio-cultural determinants of physical activity in adolescents]. AB - Physical activity is important for health promotion and disease prevention. Effective physical activity programs for adolescents require a proper understanding of the determinants of activity levels. The main purpose of this paper was to review the scientific literature on determinants of physical activity among adolescents: demographic, biological (age, gender, socioeconomic status), and socio-cultural (family, peers, and physical education teachers). The review included only studies with large samples (>100 subjects) and a cross sectional design, and that used questionnaires to measure physical activity in adolescents (10-18 years). The main results and conclusions were: age is negatively associated with physical activity; boys tend to be more active than girls; higher socioeconomic status is positively associated with more physical activity; adolescents are more involved in physical activity when parents and peers also participate; physical education teachers do not influence the adolescents' level of physical activity. PMID- 18392350 TI - [Historical evolution of package inserts in Brazil]. AB - In Brazil, package inserts provide key information on pharmaceuticals. The current study analyzes the evolution of package inserts and the impact on this process by scientific research and development, globalization of information, and various health policies. The study began with a retrospective review of Brazilian health legislation until 1920, the year when the National Public Health Department was created. The analysis of documents on the evolution of health regulation in Brazil began with the Brazilian Pharmaceutical Collection-Health Rulings. The second stage of the study involved a search of standards and norms in VISALEGIS: Health Surveillance Legislation, Portal for Legislation from the National Congressional Information System and the Health Legislation System. Package inserts became an important vehicle for information in the country and underwent important regulatory changes in the latter half of the 20th century. From 1946 to 2006, the number of mandatory items increased, with more in-depth description. However, the standardization of information for medicines with the same active ingredient failed to materialize, despite its importance and the various legal initiatives in this direction. PMID- 18392351 TI - [Factors affecting access to health care institutions by the internally displaced population in Colombia]. AB - In Colombia, the on-going armed conflict causes displacement of thousands of persons that suffer its economic, social, and health consequences. Despite government regulatory efforts, displaced people still experience serious problems in securing access to health care. In order to analyze the institutional factors that affect access to health care by the internally displaced population, a qualitative, exploratory, and descriptive study was carried out by means of semi structured individual interviews with a criterion sample of stakeholders (81). A narrative content analysis was performed, with mixed generation of categories and segmentation of data by themes and informants. Inadequate funding, providers' problems with reimbursement by insurers, and lack of clear definition as to coverage under the Social Security System in Health pose barriers to access to health care by the internally displaced population. Bureaucratic procedures, limited inter- and intra-sector coordination, and scarce available resources for public health service providers also affect access. Effective government action is required to ensure the right to health care for this population. PMID- 18392352 TI - Assessment of human health risk from organochlorine pesticide residues in Cidade dos Meninos, Duque de Caxias, Rio de Janeiro, Brazil. AB - This article presents the results of a heath risk assessment in Cidade dos Meninos, an area contaminated with organochlorine pesticide residues located in the county (municipality) of Duque de Caxias, Rio de Janeiro State, Brazil. The Agency for Toxic Substances and Disease Registry (ATSDR) risk assessment methodology was applied. The following organochlorine compounds exceeded the established limits and were considered target pollutants: HCH and its isomers (in eggs, milk, and soil); DDT and its metabolites; trichlorobenzenes; trichlorophenols; dioxins and furans (eggs and soil). For all the substances studied (with the exception of trichlorophenols), the estimated doses exceeded the minimum risk levels for chronically exposed children and adults. According to the ATSDR classification, the area was considered an "urgent public health hazard" because of its high and serious exposure to substances that are dangerous to human health. PMID- 18392353 TI - [The hospital bed referral center under the Unified National Health System in Belo Horizonte, Minas Gerais State, Brazil: the case of hospitalization for ischemic heart disease]. AB - The hospital admissions center in Belo Horizonte, Minas Gerais State, Brazil, aims to assure fast, timely, and equitable access to hospitalization services through the Unified National Health System. However, many patients are admitted directly to the hospitals, without going through the admissions center. This study compared the characteristics of hospitalizations in Belo Horizonte in 2002 according to type of access. All admissions for acute myocardial infarction and acute coronary disease were included. Of 3,705 admissions, 24.9% were processed through the hospital admissions center and 75.1% through direct access. Direct hospitalizations were more common as compared to processing by the hospital admissions center for patients>70 years), those with presumptive diagnosis of acute coronary disease, to the surgical department, and on weekends. Admissions via the hospital admissions center were more common than direct access for patients residing outside Belo Horizonte, at non-public hospitals, and for the intensive care unit. Length-of-stay also varied according to type of access. The results confirm differences in the characteristics of admissions according to the two types of access. PMID- 18392354 TI - [Assessment of knowledge and preventive attitudes concerning visceral leishmaniasis in Belo Horizonte, Minas Gerais State, Brazil]. AB - The main objective of this study was to evaluate knowledge concerning visceral leishmaniasis and attitudes used to prevent the disease in Belo Horizonte, Minas Gerais State, Brazil, in 2006. A case-control study was conducted, with home visits and a questionnaire. The odds ratio was calculated, comparing 82 cases of human visceral leishmaniasis in 2004 and 164 controls (neighbors of cases). The disease was more frequent in children (OR=109.77). Visceral leishmaniasis was 2.57 times more likely in males than in females. Overall schooling level was low (68.3% of subjects had not completed secondary school). Half of the cases did not know what visceral leishmaniasis was, and only 1.2% could identify the vector. Having basic knowledge of visceral leishmaniasis reduced the odds of acquiring the disease by 2.24 times. Keeping the house clean and taking pet dogs to the veterinarian reduced the risk by 1.94 times. In Belo Horizonte, the population's level of knowledge on visceral leishmaniasis is superficial, and preventive attitudes are inaccurate. PMID- 18392355 TI - [Is there any relationship between rural environmental factors and reproductive health in the Pampa Humeda in Argentina?]. AB - The relationship between environmental factors and health is well known. Rural environmental influences on reproductive health have been properly proved, both in animals and humans. In Latin America, few studies have been conducted in this area. The current project is based on the description of relationships between reproductive health and environmental factors in rural populations, characterized by specific environmental characteristics. Three variables were evaluated: male to-female birth ratio, male urogenital malformations (cryptorchidism and hypospadias), and endocrine-related cancer incidence. Five rural communities in the Pampa Humeda in Argentina were selected, and the data were compared to the national mean. Biomedical data and environmental risk factors were correlated through a geographic information system. The ratio of male to female births did not show any differences. Malformations showed very significant differences. Endocrine-related cancers showed higher incidence rates compared to the national mean, particularly in some communities. In conclusion, there is a relationship between environmental factors and reproductive health conditions in this region. PMID- 18392356 TI - [Religion and psychiatric disorders in patients admitted to a university general hospital]. AB - In order to evaluate the prevalence of psychiatric disorders in a Brazilian general hospital and their association with religious denomination and religiosity, 253 inpatients were interviewed. A socio-demographic questionnaire and an instrument for diagnosis of mental disorders (MINI-Plus) were applied. Distribution of religious denominations was: Catholic 63.2% (n=177), Evangelical Protestant 20.4% (n=57), Spiritist 4.3% (n=12), traditional Protestant 2.3% (n=8), and "no religion" 7.5% (n=21). Degree of religiosity was: very religious 43.2% (n=116), religious 46.9% (n=129), hardly religious 9.8% (n=27), and not at all religious 1.1% (n=3). Evangelical (Pentecostal) religious affiliation and frequent attendance at worship services were associated with fewer alcohol problems. Membership in an Evangelical (Pentecostal) church may thus have an inhibitory effect on alcohol dependence or abuse. Intensity of religiosity was moderately associated with overall prevalence of disorders, especially bipolar disorder. It is reasonable to conclude that extreme situations (very intense versus very limited religious participation) are related to this finding, associating both an exacerbated pursuit of religion and alienation from it with altered mental states. PMID- 18392357 TI - Cultural significance of primary teeth for caregivers in Northeast Brazil. AB - This anthropological study critically evaluates Brazilian caregivers' symbolic production and significance of their malnourished offspring's primary teeth, as well as their own, and describes popular dental practices. From January to June 2004, ethnographic interviews of 27 poor, low-literacy mothers were conducted at a public Malnutrition Treatment Center in Fortaleza, Ceara State. Participant observation of clinical pathways and home environments supplemented the data. Content analysis was performed. Results confirm that primary teeth are imbued with cultural significance in Northeast Brazil. Mothers examine mouths, perceive signs of decay, associate primary with permanent tooth healthiness, identify ethnodental illnesses, seek assistance, and perform rituals with exfoliated teeth. The mother's motivation to care for primary teeth is sparked by her memories of past toothache and attempts to avoid stigma and discrimination. Social determinants, not mothers' beliefs or behaviors, are the most critical obstacles to effective dental care. Legitimizing lay knowledge and empowering caregivers and children can improve oral health in Northeast Brazil. PMID- 18392358 TI - [Assessment of the Family Health Program in municipalities in Santa Catarina State, Brazil]. AB - This evaluative study aimed to classify the municipalities (counties) in the State of Santa Catarina, Brazil, as to implementation of the Family Health Program, using three categories (satisfactory, intermediate, and unsatisfactory) and two different periods (2001 e 2004). The indicators were coverage, evidence of change in the healthcare model, and impact. According to the results, coverage increased significantly from 2001 to 2004 (p<0.0001). Evidence of change in the healthcare model and impact did not change significantly. In 2001, 48% of the municipalities showed an unsatisfactory situation, as compared to 45% with intermediate and 7% satisfactory performance, while in 2004 the figures were 29%, 58%, and 13%, respectively. Improvement in classification from 2001 to 2004 was statistically significant (p=0.0061). Analysis of the data as a whole showed that changes in the final classification resulted basically from the coverage indicator, demonstrating effective expansion in access. However, these indicators did not show significant changes in the healthcare model during the period analyzed. PMID- 18392359 TI - Diet and cancer in Northeast Brazil: evaluation of eating habits and food group consumption in relation to breast cancer. AB - A hospital-based case-control study was performed from August 2002 to November 2003 in Northeast Brazil. Eighty-nine women were recruited with histologically confirmed breast cancer (age 30-80 years), matched for age with 94 controls. Food consumption of cases and controls was evaluated by foods and food groups, categorized in consumption tertiles. Odds ratio (OR) and 95% confidence intervals (95%CI) were obtained through unconditional logistic regression. Intake of fruits and juices, beans, and dairy products showed a strong association with reduced risk of breast cancer. Consumption of red and fried meat was positively associated with risk of breast cancer (red meat-OR=4.30; 95%CI: 1.74-10.67; p for trend=0.00). No association was observed in vegetable and sausage meat groups and breast cancer. Red and fried meat may be risk factors, and intake of fruit, beans, and dairy products may protect against breast cancer. PMID- 18392360 TI - [Drinking-and-driving prevalence in Belo Horizonte, Minas Gerais State, Brazil]. AB - Problems resulting from alcohol consumption by drivers have been studied worldwide, and epidemiological research points to high injury and death rates related to drinking-and-driving. However, equivalent data are limited in Brazil. In this study, 913 drivers were stopped on public roads with heavy traffic and high concentrations of bars, restaurants, and nightclubs in Belo Horizonte, Minas Gerais State, and asked to answer a questionnaire and submit to an active breathalyzer test. The study was done in December 2005 and December 2006. The study adopted the internationally accepted sobriety checkpoint method. In the sample, 38.0% of drivers showed some trace of alcohol in their exhaled air, and 19.6% were at or above the legal limit (0.6 g/l). These figures were five times those found in similar surveys in other countries. The findings suggest a critical drinking-and-driving problem in Belo Horizonte (and probably elsewhere in Brazil) and the need for on-going research, the development of specific public policies to deal with the problem, and effective enforcement of the existing law. PMID- 18392361 TI - Premenstrual symptoms and syndrome according to age at menarche in a 1982 birth cohort in southern Brazil. AB - Premenstrual symptoms and syndrome were studied in young women who have been followed since birth. Data were collected on the intensity of four symptoms: irritability, anxiety or stress, depressed mood, and affective lability. Premenstrual syndrome was defined according to intensity of symptoms. Association between age at menarche and premenstrual symptoms and syndrome were investigated through Poisson regression. Adjusted analysis was conducted, controlling for possible confounding factors. The symptoms most frequently reported by the women from the 1982 Pelotas, Rio Grande do Sul, Brazil, birth cohort who were interviewed in 2004-2005 were: irritability (52.3%) and anxiety (40.2%). The prevalence rates for moderate and severe premenstrual syndrome were 13.4% and 5.8%, respectively. Mean age at menarche was 12.4 (+/-1.5) years. Prevalence rates for symptoms and premenstrual syndrome were higher in women whose age at menarche was less than 11 years, but this difference was not statistically significant. Information on symptoms and premenstrual syndrome is scarce in other studies. PMID- 18392362 TI - Life expectancy among elderly Brazilians in 2003 according to different levels of functional disability. AB - The aim of the present study was to estimate disability-free life expectancy for the Brazilian elderly in 2003, by gender and age, based on different concepts of functional disability. The Sullivan method is used to combine the period life tables from the Brazilian Institute of Geography and Statistics (IBGE, 2003) and the prevalence of functional disability according to the 2003 National Sample Household Survey (PNAD 2003). The main results of the study indicate that at age 60, Brazilian men can expect to live 19 years, 39% with mild, 21% with moderate, and 14% with severe functional disability, respectively. At the same age, Brazilian women can expect to live 22 years: 56% with mild, 32% with moderate, and 18% with severe functional disability. PMID- 18392363 TI - Complete treatment of uncertainties in a model for dengue R0 estimation. AB - In real epidemic processes, the basic reproduction number R0 is the combined outcome of multiple probabilistic events. Nevertheless, it is frequently modeled as a deterministic function of epidemiological variables. This paper discusses the importance of adequate treatment of uncertainties in such models. This is done by comparing two methods of uncertainty analysis: Monte Carlo uncertainty analysis (MCUA) and the Bayesian melding (BM) method. These methods are applied to a model for the determination of R0 of dengue fever based on entomological parameters. The BM was shown to provide a complete treatment of the uncertainties associated with model parameters. In contrast to MCUA, the incorporation of uncertainties led to realistic posterior distributions for parameter and variables. The incorporation, by the BM, of all the available information, from observational data to expert opinions, allows for the constructive use of uncertainties generating informative posterior distributions for all of the model's components that are coherent as a set. PMID- 18392364 TI - [Family Health Program implementation in municipalities in Mato Grosso State, Brazil]. AB - This article analysis some key aspects in the implementation of the Family Health Program (FHP): results; conditions; and institutional mechanisms; flow and regularity of funding; organizational structures; and human resources availability and training. The study was conducted in seven municipalities (counties) in the State of Mato Grosso, Brazil, and used secondary data as well as primary data from interviews with different stakeholders. The research design was evaluative, using a quantitative/qualitative analysis. The results showed: varying stages in the implementation process, different FHP models, and adaptation of organizational structures; high level of human resources availability, except for nurse assistants; availability of financial resources, with some difficulties in their flow; and other institutional factors that hinder or facilitate the micro-implementation process in the municipalities. PMID- 18392365 TI - Size at birth and height in early adolescence: a prospective birth cohort study. AB - The aim of this study was to evaluate the association between size at birth (birthweight and birth length) and height in early adolescence in a prospective birth cohort study in Pelotas, Rio Grande do Sul, Brazil. Interviews were carried out in 1993, including measurements of birthweight and length, and in 2004-2005, including measurements of weight and height. This analysis includes 4,452 individuals, with a mean age of 11.3 years (standard deviation-SD=0.3), representing a follow-up rate of 87.5%. Mean height at 11 years was 145.8 cm (SD=7.9), or 144.9 cm (SD=7.7) in boys and 146.8 cm (SD=7.9) in girls. Birthweight and birth length were positively associated with height in early adolescence in the crude analysis, but after adjustment for confounding and for each other, only the effect of birth length was still significant. A one z-score increase in birth length was associated with a 1.63 cm increase in height at 11 years. The present study shows that birth length is a strong predictor of later height, while the effect of birth weight disappears after adjustment for birth length. PMID- 18392366 TI - Functional performance of the elderly in instrumental activities of daily living: an analysis in the municipality of Sao Paulo, Brazil. AB - The changes that occur during aging are accompanied by limitations in functional performance that impair elderly people's social participation and consequently harm their quality of life. In the SABE Study (Sao Paulo, Brazil), the results indicated relationships between life expectancy without disabilities and demographic and social variables. This article sought to evaluate these and other variables together as explanatory factors for the presence or absence of difficulties in instrumental activities of daily living (IADL), directly related to the possibility of more efficient community participation. Five of the eight questions relating to IADL that were applied to the sample of 2,143 elderly individuals in the municipality of Sao Paulo in 2000 were considered. Odds ratios were calculated using logistic regression. All the socio-demographic variables analyzed (age, income, ethnicity, schooling, and gender) were significant, with the exception of "sufficient money". All the variables in the "health" group (one or two or more diseases reported, and depression) were also significant. The two other variables that entered the model were physical activity and alcohol consumption. PMID- 18392367 TI - Mortality, TB/HIV co-infection, and treatment dropout: predictors of tuberculosis prognosis in Recife, Pernambuco State, Brazil. AB - This non-concurrent cohort study aims to identify predictors of tuberculosis mortality in a large population database in Brazil. Tuberculosis, death, and TB/HIV cases were validated respectively from the tuberculosis surveillance (SINAN/TB), mortality (SIM), and SINAN/AIDS databases for a five-year period. Analysis included proportional hazard models with relative risk estimates. Out of 5,451 individuals reported with tuberculosis, 320 (5.9%) died (incidence and mortality rates of 98.6 and 12.2/100 thousand inhabitants, respectively). After adjustment, relative risk of dying from tuberculosis was 9.8 for individuals>50 years of age; 9.0 for TB/HIV co-infection; 3.0 for mixed TB clinical presentation; and 2.0 for treatment dropout. In the multivariate model, using cases with HIV/AIDS, all adjusted predictors lost significance except mixed clinical presentation (RR 1.9; 1.1-3.1). TB/HIV co-infection is an important predictor of TB mortality. However, among individuals without HIV/AIDS, mortality is still highly associated with older age, mixed clinical forms, and treatment dropout. PMID- 18392368 TI - Revised criteria for the assessment and interpretation of occlusal deviations in the deciduous dentition: a public health perspective. AB - Secondary data from a randomized sample of Brazilian preschool children were analyzed to evaluate the prevalence of occlusal deviations in the deciduous dentition that can adversely affect the permanent dentition, using revised criteria. Overjet and overbite used cut-off points described in the literature to exclude mild malocclusion cases. Overjet>3 mm and overbite>3 mm were present in 16% and 7% of the children, respectively. On the sagittal plane, only bilateral deviations were considered: molar relationship in distal step (9.7%) and mesial step (6%) and canine relationship class 2 (11%) and class 3 (2.9%). Regarding other occlusal deviations, severity criteria were not reported in the literature. Crude prevalence rates of openbite (27.9%), posterior crossbite (11.3%), and crowding in the maxillary arch (7%) and mandibular arch (11.3%) were recorded. The assessment of malocclusion in the deciduous dentition should focus on the severity of occlusal deviations for the identification of cases and non-cases that are relevant to public health. Need for further consensus and improvement in the interpretation of epidemiological data on malocclusion related to this developmental stage is emphasized. PMID- 18392369 TI - [The Family Health Program in Brazil and the adequacy of its coverage indicator]. AB - This study aimed to evaluate the adequacy of the Family Health Program's (FHP) coverage indicator, calculated by using the estimated number of people served by the FHP team, obtained from a national standard. The study included all the Brazilian counties (municipalities) that adopted the FHP strategy from 1998 to 2006. The analytical methods were Pearson's correlation, Lin's concordance correlation coefficient, and the Deming regression model. The results showed that at the beginning of the observation period, the coverage indicator did not express the size of the program's population of beneficiaries, when comparing the enrolled population, the number of families under follow-up, and the number of live births recorded. Gradually, and especially beginning in 2000, the indicator began to represent a good estimate of FHP coverage, although the trend differed according to the size of the municipalities (it was more evident among the larger municipalities). The study emphasizes the importance of defining coverage of actions and services in order to orient health program implementation. PMID- 18392370 TI - [Factors associated with interpersonal continuity of care: a population-based study]. AB - This study focused on the prevalence of interpersonal continuity of care and its determinants. A cross-sectional population-based study was conducted with 3,133 individuals 20 years or older in Pelotas, Rio Grande do Sul State, Brazil. Multivariate analysis used Poisson regression, with the first level representing socioeconomic and demographic variables and the proximal level including health care and health needs variables. Prevalence of interpersonal continuity of care was 43.7% (95%CI: 42.0-45.5). Female gender, higher age, higher income, consultation during the previous year, report of chronic disease, and consultation outside the public health system showed higher interpersonal continuity of care. Among individuals that consulted in public primary health care services, female gender, age, and the Family Health Program were associated with interpersonal continuity of care. Some vulnerable groups (low-income and users of the public health system) showed low prevalence of the outcome. PMID- 18392372 TI - [Quality of life and perception of illness among individuals with high blood pressure]. AB - This study aimed to investigate quality of life in individuals with arterial hypertension and to associate the results with the patients' perceived severity of illness. The sample included 113 patients with arterial hypertension followed up in a clinic in Fortaleza, Ceara State, Brazil, from May to August 2002, using an international scale for assessing quality of life, the SF-36. Of the entire sample of patients, 77% were women, a large proportion were over 50 years of age, living with family, and with some kind of occupation. Assessment with the SF-36 showed compromised quality of life in different domains, although patients viewed their illness as not serious, and curable. These circumstances can interfere in follow-up of their illness over time, which can in turn jeopardize their quality of life. In conclusion, use of the SF-36 to assess individuals with arterial hypertension produced significant results, indicating generally compromised quality of life. According to these results, hypertension jeopardizes the patients' social and physical dimensions, but this situation can be changed by the provision of qualified services. PMID- 18392371 TI - [A descriptive profile of the canine population in Aracatuba, Sao Paulo State, Brazil, from 1994 to 2004]. AB - From 1994 to 2004, the canine population in Aracatuba, Sao Paulo State, Brazil, suffered two major canine zoonoses: rabies and visceral leishmaniasis. Changes in the dog population during this period were evaluated using canine census data from 1994 and 2004 and the results of blood samples for diagnosis of canine visceral leishmaniasis in 1999. The ratio of dogs per 10 inhabitants varied from 1.7 in 1994 to 2.0 in 1999 and 1.8 in 2004. The percentage of puppies less than 1 year of age increased from 20% to 32.5%, and the number of euthanized dogs also increased after 1999, when visceral leishmaniasis began to appear. The number of dogs and percentage of puppies varied between different areas of the city, and neighborhoods with a higher percentage of young animals showed more cases of both human and canine leishmaniasis. This result may be due to control measures applied in these areas in response to cases of human and canine visceral leishmaniasis, but the increase in the younger canine population can be accompanied by increased susceptibility in these animals, thus favoring maintenance of the disease in the area. PMID- 18392374 TI - The university and its hierarchical layers. PMID- 18392373 TI - Anomalous origin of the right coronary artery: diagnosis and treatment. AB - We report the case of a rare anomaly of the right coronary artery (RCA) arising from the left aortic sinus, having a single ostium with the left coronary artery, associated with documented episodes of inferior ischemia, in which surgical treatment with a right internal thoracic artery to RCA bypass graft and the corresponding ligation provided greater stability to the coronary blood flow and good clinical progress. PMID- 18392375 TI - Twiddler syndrome. PMID- 18392376 TI - Prehospital thrombolysis in AMI: a feasible alternative to Brazil? PMID- 18392377 TI - Evaluation of heart rate variability in trained and sedentary climacteric women. AB - BACKGROUND: Changes in autonomic cardiac function are frequent during menopause, and various methods have been used to understand and minimize them. OBJECTIVE: To study the interference of dynamic aerobic physical activity on heart rate variability (HRV) in climacteric women. METHODS: Cross-sectional study that analyzed HRV in 15 menopausal women (mean age 56.8+/-4.9 years) who had participated in physical training (one-hour walks, 3 times a week) for at least two years (active group), and 15 menopausal women (mean age 56.5 +/- 3.7 years) who were sedentary (sedentary group). None of the volunteers received hormonal replacement therapy. HRV data were compared between the groups by means of the Mann Whitney U Test. RESULTS: There were significant differences both in the frequency and time domains of the following variables of HRV (in medians) for the active e sedentary groups, respectively: total power (22,626.50 ms(2) and 4,432.10 ms(2)), low frequency component (741.20 ms(2) and 131.70 ms(2)), high frequency component (668.90 ms(2) and 131.70 ms(2)), standard deviations of RR intervals (51.60 ms and 22.50 ms), square root of the sum of squares of differences between the normal RR intervals (35.30 ms and 15.90 ms), and percentage of normal adjacent RR intervals greater than 50 ms (6.6% and 0.2%). CONCLUSION: The study suggests that aerobic training may have afforded a significant improvement in the autonomic cardiac function of the menopausal women in the active group, and may be a useful option for preserving this functional condition without the need for hormonal replacement therapy. PMID- 18392378 TI - Influence of external temporary biatrial pacing on the prevention of atrial fibrillation after coronary artery bypass without extracorporeal circulation. AB - BACKGROUND: Atrial fibrillation is the most common complication after myocardial revascularization, and it increases morbidity/mortality. OBJECTIVE: The purpose of this prospective randomized study was to test the hypothesis that temporary biatrial pacing is effective in reducing the incidence of postoperative atrial fibrillation after myocardial revascularization. METHODS: Ninety-eight non consecutive patients who had undergone off-pump myocardial revascularization received two temporary electrodes attached to the right and left atria, which were connected to either pair of atrial pacemaker electrodes, in addition to the leads implanted in the right ventricle. Two groups of patients were randomized (control: 49 patients with no biatrial pacing; therapeutic: 49 patients with biatrial pacing). The variables of interest were atrial fibrillation (present or absent) and length of hospital stay. RESULTS: The incidence of atrial fibrillation was 36.73% in the control group and 14.29% in the therapeutic group (p=0.0194). Length of hospital stay was 7.00 +/- 2.82 days for patients with no atrial fibrillation (n=73) and 9.20 +/- 2.87 days for patients with atrial fibrillation (n=25) (p=0.0001). Age was an important predictor of arrhythmia and ranged between 62.34 +/- 9.00 years in the group with no atrial fibrillation and 67.20 +/- 7.42 years in the group with atrial fibrillation (p=0.0170). CONCLUSION: Compared to controls, prophylactic temporary biatrial pacing is effective in preventing atrial fibrillation. Hospital stay was longer for patients who developed postoperative atrial fibrillation, and age was an important predictor for the development of arrhythmia. PMID- 18392379 TI - Serum levels of interleukin-6 (Il-6), interleukin-18 (Il-18) and C-reactive protein (CRP) in patients with type-2 diabetes and acute coronary syndrome without ST-segment elevation. AB - BACKGROUND: Atherosclerosis is an inflammatory disease, and serum levels of inflammatory markers such as interleukin 6 (IL-6), interleukin 18 (IL-18) and C reactive protein (CRP) are used to evaluate patients with coronary artery disease. In patients with type-2 diabetes, atherosclerosis is related to a larger number of events such as myocardial infarction and death, when compared with patients without diabetes. OBJECTIVE: To evaluate the inflammatory response in patients with diabetes and acute events of coronary instability. METHODS: Two groups of patients were primarily selected. The first group was comprised of diabetic outpatients with stable angina (D-CCS) and presence of coronary artery disease on coronary angiography (n=36). The second group was comprised of diabetic patients seen in the emergency room with acute coronary syndrome (D-ACS) without ST-segment elevation (n=38). Non-diabetic patients with ACS (n=22) and CCS (n=16) comprised the control group. Serum levels of CRP, IL-6 and IL-18 were determined using nephelometry (CRP) and ELISA (IL-6 and IL-18) techniques. RESULTS: Higher serum IL-6 levels were found in diabetic or non-diabetic patients with ACS than in the group with CCS. On the other hand, diabetic patients with ACS had higher CRP levels in comparison with the other groups. Serum IL-18 levels were not significantly different among the patients studied. CONCLUSION: our findings suggest a more intense inflammatory activity in patients with coronary instability. This inflammatory activity, as measured by CRP, seems to be even more intense in diabetic patients. PMID- 18392380 TI - Cost-effectiveness of prehospital versus inhospital thrombolysis in acute myocardial infarction. AB - BACKGROUND: There is evidence that prehospital thrombolysis improves the outcome in ST-elevation myocardial infarction (STEMI). OBJECTIVE: This study aimed at comparing the cost-effectiveness of prehospital compared to inhospital thrombolysis for STEMI from the National Health System perspective. METHODS: A decision-analytic model was used to compare these two strategies. The study endpoint was life-years gained. Resource use and costs were estimated from the National Health System perspective. The Decision tree and Markov Model were constructed using the results of published clinical trials. Costs were expressed in Reais (R$), for the year 2005. RESULTS: For a time horizon of 20 years, prehospital thrombolysis had an average life expectancy of 11.48 years and inhospital thrombolysis had an average life expectancy of 11.32 years. Costs were R$ 5,640 for prehospital thrombolysis and R$ 5,816 for inhospital thrombolysis. Inhospital thrombolysis led to an additional cost of R$ 176 per patient. Pre hospital thrombolysis led to additional 0.15 years of life-expectancy gain compared with inhospital thrombolysis. CONCLUSION: This model suggests that, from the National Health System perspective, implementing prehospital thrombolysis for ST-elevation myocardial infarction (STEMI) may lead to extra survival and fewer costs when compared to inhospital thrombolysis. PMID- 18392381 TI - Determination of microalbuminuria in hypertensive patients and in patients with coronary artery disease. AB - BACKGROUND: The normal 24-hour albumin excretion rate is of 20 mg. A persistent rate of 30 to 300 mg/day is called microalbuminuria and is related to a higher prevalence of cardiovascular disease. OBJECTIVE: 1) To determine the prevalence of microalbuminuria in a group of hypertensive patients and in a group of patients with coronary artery disease; 2) To determine the relationship between the presence of microalbuminuria and hypertension, diabetes mellitus, dyslipidemia, smoking and obesity. METHODS: The presence of microalbuminuria in a group of hypertensive patients (73 individuals) and in a group of patients with coronary artery disease (39 individuals) was determined and compared with a control group (43 individuals). Microalbuminuria was defined as an albumin/creatinine ratio higher than 30 and lower than 300 in a spot morning urine sample. The chi-square test and the Fishers exact test were used in the statistical analysis. RESULTS: Microalbuminuria was present in 9.5% of the hypertensive individuals and in 33% of the patients with coronary artery disease, and was absent in individuals of the control group. When the occurrence of microalbuminuria was analyzed according to the different clinical parameters, regardless of the group involved, a statistically significant correlation was found with age, diabetes and dyslipidemia. CONCLUSION: 1) The prevalence of microalbuminuria in hypertensive individuals is high, and is even higher in patients with coronary artery disease; 2) There is a correlation of the presence of microalbuminuria with age, diabetes and dyslipidemia. PMID- 18392382 TI - Intermittent doses of statin in hemodialysis patients with spontaneous low LDL cholesterol levels. AB - BACKGROUND: Mortality in dialysis patients remains high and is due mainly to cardiovascular causes. Inflammation has a role in the genesis of accelerated atherosclerosis, vascular calcification, malnutrition and anemia, and a huge impact on the survival of these patients. The pleiotropic effects of statins can be a therapeutic option for reducing chronic inflammatory processes of patients undergoing hemodialysis. OBJECTIVE: To evaluate the effects of low doses of simvastatin on inflammatory markers, hematimetric and nutritional parameters of patients undergoing hemodialysis. METHODS: Clinically-stable patients undergoing hemodialysis were classified according to their baseline LDL-cholesterol levels in two groups: those with levels below 100mg/dl (Group 1) and those with levels equal to or greater than 100mg/dl (Group-2), and were treated with simvastatin during eight weeks. Group 1 received 20mg only after each session of hemodialysis (intermittent dose), whereas Group 2 received 20mg/daily. Laboratory data, erythropoietin resistance index and nutritional parameters were obtained before and after treatment. RESULTS: A significant and equivalent reduction in C reactive protein levels in both groups was observed (35.97+/-49.23% vs 38.32+/ 32.69%, p=0.86). In group 1, there was also a tendency towards reduced resistance to erythropoietin (228.6+/-16.2 vs 208.9+/-16.2, p=0.058) and improvement of hematimetric parameters (hematocrit: 33.1+/-5.9% vs 36.1+/-4.5%, p=0.021). CONCLUSION: Intermittent doses proved to be as effective as the usual dose in reducing C-reactive protein levels and resistance to erythropoietin, besides improving the hematimetric parameters, indicating an important reduction of the cardiovascular risk evaluated by these parameters. PMID- 18392383 TI - Radiofrequency ablation of paroxysmal atrial fibrillation: factors determining long-term clinical efficacy. AB - BACKGROUND: Most of the studies on this subject have reported predictors of recurrence of atrial fibrillation after catheter ablation with relatively short follow-up periods. OBJECTIVE: To retrospectively evaluate predictors of long-term recurrence of paroxysmal atrial fibrillation (AF) in patients undergoing pulmonary vein isolation following one single procedure. METHODS: The authors studied a total of 139 patients (102 men; mean age of 55 +/- 12 years) undergoing radiofrequency ablation using the ostial or extra-ostial techniques for left atrial approach, combined or not with cavotricuspid isthmus ablation (CTI). Pre, intra and post-ablation variables were evaluated using univariate and multivariate analyses to determine the predictors of recurrence of AF after one procedure. RESULTS: After a 33 +/- 12-month follow-up, we observed that a longer time of history of AF, use of more antiarrhythmic drugs, and recurrence of AF within 60 days post-procedure increased the risk of long-term recurrence of AF. On the other hand, the association of atrial flutter and concomitant CTI ablation reduced the risk of recurrence of AF. CONCLUSION: Clinical variables such as time of history of AF and a larger number of antiarrhythmic drugs already used influenced the outcomes of catheter ablation. In patients with associated atrial flutter, simultaneous CTI block significantly reduced the long-term recurrence of atrial fibrillation. PMID- 18392384 TI - Hospital admission and hospital death associated to ischemic heart diseases at the National Health System (SUS). AB - BACKGROUND: Analyses on mortality rate due to diseases when outcome depends on proper, timely medical intervention may point out the vulnerabilities and inequity associated to health care access. Ischemic heart diseases will act as models for such assessment. OBJECTIVE: The present study investigates factors associated to the hospital death rate of patients admitted to hospital due to acute myocardial infarction (AMI) and heart failure (HF), and whether admittance through Belo Horizonte Municipal Health Office (SMSA-BH) Admission Center (AC) was in any way associated to hospital death after adjustment of relevant factors. METHODS: Data obtained from the Hospital Admission Authorizations (AIH) and requests for hospital beds at SMSA data base on latest hospital admissions based on AMI or HF diagnostic hypotheses. Multivariate analysis was conducted to investigate risk factors for hospital death. RESULTS: No association was found between hospital admittance access and hospital death risk from those causes. Multivariate analysis showed higher death risk for 60 and 60+ year-old patients (OR=2.9), AMI diagnostic hypothesis (OR=3.0), the need for ICU care (OR=1.6), females (OR=1.4), surgery type (OR=1.9), and public health service hospital (OR=3.5). Hospital admissions due to AMI on weekends also showed higher death risk for death (OR=1.7). CONCLUSION: Further investigation is necessary in order to evaluate the kind of medical assistance provided on weekends at public hospitals. Other hospital factors are to be taken into account, as well as patients and assistance procedures, as subsidies for proposals to ensure higher equity and quality standard for public health services. PMID- 18392385 TI - Validation of the short form of the Spanish hypertension quality of life questionnaire (MINICHAL) for Portuguese (Brazil). AB - BACKGROUND: The assessment of quality of life has been considered an essential parameter in understanding the impact of arterial hypertension. OBJECTIVE: To translate into portuguese, cross-culturally adapt and validate the questionnaire specific for the assessment of quality of life known as "Mini Cuestionario de Calidad de Vida en Hipertension Arterial" (MINICHAL). METHODS: Two independent translations of MINICHAL were made into brazilian portuguese. Later, these two translations were harmonized yielding a version that was back-translated into the source language. This version was revised by a committee of judges and the new version was tested in a pilot study. After the cross-cultural adaptation, the final version of the instrument was administered to a sample of 300 patients. The psychometric properties of the questionnaire such as reliability and construct validity were analyzed. The internal consistency of the instrument was measured by Cronbachs alpha coefficient. RESULTS: In the brazilian version of MINICHAL, Cronbachs alpha coefficients of internal consistency reliability were 0.88 for the Mental Status domain and 0.85 for the Somatic Manifestations domain. As to content validity, the judges assessment attained a high level of agreement (75.44%). The factor analysis confirmed both domains, with differences in one item which was included in factor 2. The control group presented significant differences relative to hypertensive patients (t=4.86, gl=276.8, p< 0.001). CONCLUSION: The brazilian version of MINICHAL was successfully validated and represents a useful and reliable instrument for assessing the quality of life of brazilian hypertensive patients. PMID- 18392386 TI - Heart failure with preserved ejection fraction and systolic dysfunction in the community. AB - BACKGROUND: In developed countries, heart failure with preserved ejection fraction (HFpEF) is more prevalent than heart failure with reduced ejection fraction (HFrEF) in the community. However, it has not been completely established if this fact is also observed within our community. OBJECTIVE: To determine the most prevalent form of heart failure (HFpEF or HFrEF) and whether the prevalence of HFpEF is higher in the community. METHODS: This is a cross sectional study conducted with patients clinically diagnosed with HF who were seen in community-based health care centers from January to December 2005. Echodopplercardiograms were performed for all patients. The form of HF was stratified according to the presence of abnormalities and the shortening fraction observed on the echodopplercardiogram. RESULTS: The study evaluated 170 patients (61.0 +/- 13.3 years of age), most of them women and elderly. HFpEF was the more prevalent form of HF (64.2%, p<0.001), affecting mostly elderly women (62%, p = 0.07), whereas the opposite condition, HFrEF, was observed mostly in elderly men (63.6%, p = 0.07). Patients with no HF represented one-third of the cases (27.6%). HFrEF patients had more lower-limb edema, coronary disease, diabetes, chronic renal failure, higher Boston scores and hospital readmissions. Use of alcoholic beverages and smoking were also more common among HFrEF patients. CONCLUSION: HFpEF is the most prevalent form of HF in the community especially among elderly women, whereas HFrEF affects mostly elderly men and is associated with greater clinical severity, main risk factors and no changes in lifestyle. Despite the signs and symptoms of HF, this condition was not confirmed for one third of the cases. PMID- 18392387 TI - Functional behavior of patients with conventional pacemakers undergoing cardiac resynchronization. AB - BACKGROUND: Cardiac resynchronization therapy (CRT) is an efficient treatment for patients with heart failure (HF), severe ventricular dysfunction and intraventricular block. Conventional pacemakers (CPM) implanted in the right ventricular apical area cause alterations in the normal sequence of cardiac activation similar to those induced by LBBB (left bundle-branch block). Therefore, patients with CPM and advanced HF could be candidates to undergo CRT, but as only small numbers of patients have been evaluated so far, definitive conclusions are lacking. OBJECTIVE: To assess the clinical and functional outcome of cardiac resynchronization therapy (CRT) in patients with conventional pacemakers. METHODS: Patients with CPM, who were in NYHA HF functional class III/IV class refractory to drug therapy, and left ventricular ejection fraction (LVEF) ) <35% underwent CRT. Patients clinical-functional behavior was assessed prospectively six months after the procedure. The improvement of one HF functional class was set as an effective response to the procedure. The following was assessed: QRS duration (ECG), diastolic diameter (LVDd), left ventricular systolic diameter (LVSd) and LVEF seen on the echocardiogram. For the statistical analysis, Students paired t test and Spearmans correlation were used. RESULTS: Twenty-nine patients (mean age 61.5) were evaluated. Of these, six were females, and chagasic cardiomyopathy was predominant. During the clinical follow-up of 22.7+/-13 months, 86.2% of the patients benefited from CRT. Within this group, the mean LVEF increased by 18% (p=0.013), QRS duration dropped by 11.8% (p=0.002) and no significant reduction in left ventricular intracavitary diameters was observed. CONCLUSION: CRT is effective for patients with CPM and advanced HF as it yields a high rate of response (86.2%), significantly improves LVEF and reduces QRS duration. PMID- 18392388 TI - Case 1/2008: three year old female patient with subvalvular, valvular, and supravalvular pulmonary stenosis. PMID- 18392390 TI - Takotsubo cardiomyopathy causing transitory ventricular dysfunction. AB - This is the report of a 74-year-old female patient with a history of systemic hypertension and peripheral vascular disease who presented acute coronary syndrome symptoms. Coronary angiography showed coronary arteries with no significant obstructions. Ventriculography and echocardiography showed akinesia in mid and apical segments; and hyperkinesia of left ventricle basal segments. Two weeks after the onset of symptoms, a new echocardiogram demonstrated normal global and regional systolic function. The uncommon, reversible pattern for systolic dysfunction and segmental compromising that gives left ventricle a Takotsubo-like shape is known today as stress cardiomyopathy. PMID- 18392391 TI - Can we believe in levosimendan? PMID- 18392392 TI - Cardioinhibitory carotid sinus hypersensitivity: prevalence and predictors in 502 outpatients. AB - BACKGROUND: Cardioinhibitory response (CIR) is defined as asystole >3 seconds in response to 5-10 seconds of carotid sinus massage (CSM). Pacemaker implantation is indicated for patients with unexplained syncope episodes and CIR. OBJECTIVE: To determine the prevalence and predictors of CIR in patients with a high prevalence of cardiovascular disease, and assess the clinical significance of CIR in patients with a history of unexplained syncope or falls. METHODS: Cross section design study. Outpatients, aged > or =50 years, referred to the electrocardiography sector of a tertiary hospital. Those with dementia, carotid bruit, and history of myocardial infarction, stroke or transient ischemic attack in the preceding 3 months were excluded. CSM was performed by a single investigator, with the patients in the supine position. CSM was applied on the right side and then on the left side during 10 seconds each time. RESULTS: 502 patients underwent CSM. CIR was present in 52 patients (10.4%; 95% CI: 7.7%-13%). Independent predictors of CIR were male gender (OR: 2.61%; CI 95%: 1.3%-5.1%), structural heart disease (OR: 3.28%; CI 95%: 1.3%-7.9%) and baseline heart rate (P<0.05). The sensitivity of the CIR to CSM in syncope evaluation was low (9.8%). Specificity was high (89.5%), being even better in women (95.3%) and in those without structural heart disease (96.2%). CONCLUSION: CIR was detected in 10.4% of the patients aged > or = 50 years. In males and in patients with structural heart disease CIR was more common. In women and patients with no apparent structural heart disease, the presence of CIR was a highly specific finding in the evaluation of syncope or falls. PMID- 18392393 TI - Sexual activity after myocardial infarction: taboo or lack of knowledge? AB - BACKGROUND: Despite its clinical and social relevance, resumption of sexual activity after an acute coronary syndrome (ACS) is a subject that is poorly addressed by healthcare providers and one that is given scant emphasis during hospitalization. Erectile dysfunction is a common complaint among cardiac patients, and these patients are in doubt regarding the safety of resuming sexual activity after a cardiovascular event. OBJECTIVE: To describe how well informed patients are in regard to acute an myocardial infarction (AMI) and what sort of guidance they receive regarding resumption of sexual activity. METHODS: A cross sectional study was performed from June to July 2005 with patients on the sixth day of AMI. A questionnaire was applied to assess not only patients' knowledge of AMI, but also their expectations regarding their resumption of sexual activity - and frequency of same - following hospital discharge. RESULTS: Ninety-six patients were included in the study (mean age 59 +/- 12); 70% of them were male and 80% had experienced their first AMI. Of these, 96% lacked any knowledge of AMI clinical manifestations and possible limitations, 63% reported an active sex life before the event, 60% were in doubt regarding their sex life following hospital discharge, and only 4% received sexual guidance from healthcare providers during their hospital stay. CONCLUSION: According to our findings, the recommendations patients receive during hospitalization are suboptimal in regard to both AMI and resumption of sexual activity. The updating of healthcare providers may change this situation. This is especially true for the nursing staff, since they spend more time with patients. PMID- 18392394 TI - ICD patients with elevated defibrillation threshold: clinical behavior and therapeutic alternatives. AB - BACKGROUND: The ideal programming of the implantable cardioverter defibrillator (ICD) shock energy should be at least 10J above the defibrillation threshold (DFT), requiring alternative techniques when the DFT is elevated. OBJECTIVE: To assess the clinical behavior of ICD patients with DFT>25J and the efficacy of the chosen therapy. METHODS: Patients who had undergone ICD implantation between Jan/00 and Aug/04 (prospective database) and presented intraoperative DFT>25J were selected. The analyzed variables were: clinical characteristics, LVEF, rescue of arrhythmic events from ICD and causes of deaths. RESULTS: among 476 patients, 16 (3.36%) presented DFT>25J. The mean age was 56.5 years, and 13 patients (81%) were men. According to the baseline cardiomyopathy, 09 patients had Chagas' disease, 04 had ischemic cardiomyopathy and 03 had idiopathic cardiomyopathy. Mean LVEF was 0.37 and amiodarone was used by 94% of the patients. Mean follow-up (FU) period was 25.3 months. DFT was higher than maximum energy shock (MES) in 2 patients and it was necessary to implant an additional shock electrode (array). It was programmed MES in ventricular fibrillation zone of ICD therapy in the other patients. In the FU, 03 patients had 67 successful appropriate shock therapies (AST). There were 05 noncardiac and 02 heart failure deaths. The patients who died showed higher DFT levels (p=0.044) without correlation with death because there wasn't unsuccessful AST. CONCLUSION: In this cohort of ICD patients, the occurrence of elevated DFT (>25J) was low, leading to alternative therapies. There was an association with severe ventricular dysfunction, although without correlation to the causes of death. PMID- 18392395 TI - Analysis of the Takeuchi procedure for the treatment of anomalous origin of the left coronary artery from the pulmonary artery. AB - BACKGROUND: Evaluation of a case series of anomalous origin of the left coronary artery from the pulmonary artery corrected with the Takeuchi procedure, analyzing their immediate and late surgical outcomes as well as the related morbidity. OBJECTIVE: To report a case series of anomalous origin of the left coronary artery from the pulmonary artery corrected with the Takeuchi procedure. METHODS: The analysis was based on data collected from the medical records of 12 patients undergoing surgical correction between 1981 and 2003. RESULTS: The mean age of the patients was 5.9 +/- 1.7 months. The predominant manifestation was heart failure, especially in the infants. The baseline electrocardiogram showed a Q wave in DI and aVL and signs of myocardial ischemia in ten cases. Echocardiogram with a pattern of dilated cardiomyopathy and mitral regurgitation was found in six patients. The mean age at surgical correction was 31.8 +/- 14.3 months. The immediate surgical mortality was 16% (two patients), from cardiac dysfunction. In the outpatient follow-up we observed that the symptoms, ischemic electrocardiographic changes, cardiac function and mitral regurgitation improved. Three cases progressed with supravalvar pulmonary stenosis during follow-up. CONCLUSION: This heart disease should be suspected in every infant presenting with heart failure. Surgical correction with the Takeuchi procedure has proven efficient, with a low mortality rate. Cardiac function returns to normal and mitral regurgitation improves after surgical correction. PMID- 18392396 TI - Influence of financial subsidy and venue on advanced cardiac life support courses, in the learning of cardiovascular emergency. AB - BACKGROUND: The success in learning of emergency depends on many factors that can be summarized as: student, instructors and course. OBJECTIVE: To evaluate the influence of financial subsidy and venue of course in learning cardiovascular emergency. METHODS: Data were analyzed regarding the courses of Advanced Cardiac Life Support (ACLS) in the period from December 2005 to December 2006. In agreement with the financial subsidy, they were divided in: group 1 - integral subsidy; group 2 - subsidy of 50%; and group 3 - without subsidy. As for the venue of the course, they were divided in: locality A - study in city with> 1 million inhabitants; and locality B - study in city with <1 million inhabitants. The practical and theoretical approval and the theoretical average were compared. RESULTS: 819 students participated in ACLS: 199 (24%) in group 1, 122 (15%) in 2 and 498 (61%) in 3. The practical and theoretical approval and the average in the theoretical exam were greater in group 3 than in other groups (p <0.05). Four hundred and eighty two (482) took the course in venue A (59%) and 337 (41%) in venue B. The practical approval was similar for both groups (p = 0.33), however the theoretical approval was greater in venue A (73% vs. 65% - p = 0.021 - OR = 1.44 and IC: 1.05 - 1.97). The theoretical average was greater in venue A (87.1 +/- 10.4 and 86 +/- 11, respectively p <0.05). CONCLUSION: The financial subsidy and venue of the course had influence in the theoretical and practical approval. PMID- 18392397 TI - Exercise testing early after myocardial infarction: comparison with echocardiography, electrocardiographic monitoring and coronary arteriography. AB - BACKGROUND: Predischarge exercise testing early after myocardial infarction is useful for risk stratification, exercise prescription, and assessment of prognosis and treatment. OBJECTIVE: The objective of this study was to compare the findings of exercise testing early after myocardial infarction with those of echocardiography, electrocardiographic monitoring (24-hour Holter monitoring) and coronary angiography. METHODS: We evaluated 60 cases (mean age of 51.42 +/- 9.34 years), of which 46 were males (77%). The symptom-limited maximal exercise test according to the Naughton protocol12 was performed between the sixth day of hospitalization and hospital discharge, with the patients on medication. During hospitalization, the patients underwent echocardiography, electrocardiographic monitoring and coronary angiography. The significance level was set at 0.05 (alpha = 5%). RESULTS: Exercise testing had a poor performance in the detection of multivessel coronary artery disease (sensitivity, 42%; specificity, 69%). No significant differences were found when the presence of ischemia on exercise test was compared with multivessel coronary disease, complex ventricular arrhythmias on electrocardiographic monitoring, and the finding of an ejection fraction lower than 60% on echocardiography (p = 0.56), as well as with the presence of multivessel lesions, complex ventricular arrhythmias on electrocardiographic monitoring and abnormal ejection fraction on echocardiography (p = 0.36). CONCLUSION: The presence of ischemia during exercise testing was associated with the occurrence of ventricular arrhythmias on electrocardiographic monitoring, with reduced ejection fraction on echocardiography, as well as with the presence of multivessel coronary lesions, which constitutes an indicator of a high coronary risk. PMID- 18392398 TI - Levosimendan in decompensated heart failure patients: efficacy in a Brazilian cohort. Results of the BELIEF study. AB - BACKGROUND: Levosimendan is a new inodilatory agent that enhances cardiac contractility via Ca(2+) sensitization and induces vasodilation through the activation of KATP/BKCa. OBJECTIVE: To study the efficacy and safety of levosimendan in a decompensated heart failure (DHF) Brazilian cohort, and in b adrenergic agonist resistant patients. METHODS: The Brazilian Evaluation of Levosimendan Infusion Efficacy (BELIEF) study was prospective, multicenter, observational and included 182 high-risk DHF patients, all of which received open label levosimendan. Primary end point was hospital discharge without additional inotropic therapy (responder). Secondary end points were changes in hemodynamics, clinical parameters, and brain natriuretic peptide (BNP). RESULTS: Mortality rate was 14.8%, and 139 of 182 patients were responders. In non responders it was 62.8%. Systolic blood pressure was a predictor of response. In b-adrenergic agonist resistant group, 55.8% were responders. Overall, 54 patients experienced at least one adverse event; most of them resolved either spontaneously or after levosimendan dose reduction. A significant improvement in quality of life was verified at 2-6 months of follow-up (p<0.0001). CONCLUSION: Our results suggest levosimendan infusion as an alternative therapy in the short term management of DHF patients. HF severity can influence the response to levosimendan treatment. Prospective studies are warranted in a Brazilian cohort including Chagas heart disease. PMID- 18392399 TI - Clinical, electrocardiographic and echocardiographic findings in significant cardiac amyloidosis detected only at necropsy: comparison with cases diagnosed in life. AB - BACKGROUND: Currently, many cases of heart amyloidosis still fail to be diagnosed. OBJECTIVE: To disclose factors related to the difficulty in attaining the diagnosis of cardiac amyloidosis. METHODS: We compared the clinical, electrocardiographic and echocardiographic data of 17 patients in whom amyloidosis was diagnosed only at the necropsy (group I) with data from 9 patients in whom the disease was diagnosed in life (group II). The quantitative variables were compared by t-test and qualitative ones by Fisher's exact test. Significance was set at p< or = 0.05. RESULTS: The two groups showed differences regarding age (group I: 75.29 +/- 11.61, group II: 58.67 +/- 11.07 years), association with other cardiac disease (group I: 52.94%, group II: 0%), low voltage at the ECG (group I: 17.65%, group II: 66.67%), and diastolic dysfunction at the echocardiogram (group I: 7.69%, group II: 62.50%). Some degree of left ventricular thickening was found in 75% of necropsy cases and 100% of controls (p=0.23), but wall thickness was lower in group I (free left ventricular wall: 1.20 +/- 0.28 cm versus 1.53 +/- 0.18 cm in group II, p=0.01). Systolic dysfunction was present in 57.89% of the cases, without significant difference between the groups. CONCLUSION: Amyloidosis is diagnosed when the clinical, ECG, and echocardiogram patterns are "typical", but most of the cases fail to be diagnosed, especially in elderly people, due to the association with other cardiac diseases, lack of diastolic dysfunction at the echocardiogram and only a slightly thickened ventricular wall. PMID- 18392400 TI - Coronary artery disease in patients with rheumatic and non-rheumatic valvular heart disease treated at a public hospital in Rio de Janeiro. AB - OBJECTIVE: to estimate the prevalence of coronary artery disease (CAD) in valvular heart disease of rheumatic (RVHD) and non-rheumatic (NVHD) etiology, assessing possible predictive factors for the presence of CAD. METHODS: This is a cross-sectional study of a series of cases obtained from a pre-defined population, wherein 1,412 patients referred for heart surgery of any etiology were evaluated. Of these, 294 primary heart disease patients aged > or =40 submitted to cinecoronary arteriography (CA) were identified and studied. RESULTS: patients with RVHD presented lower prevalence of CAD (4%) when compared to NVHD (33.61%), p<0.0001. The logistic regression analysis showed that age, typical angina-like chest pain (TACP), systemic arterial hypertension (SAH), diabetes and dyslipidemia were significantly related to CAD, and that the rheumatic etiology was not a disease determinant. Smoking and gender were clinically important in CAD, although not statistically significant. In the whole group, the Log-linear analysis showed that, regardless of the etiology, gender, age > or =55, SAH, TACP, diabetes and dyslipidemia were all related directly to CAD, with the latter three being the most important variables for the disease. CONCLUSION: the prevalence of CAD among RVHD patients is low, whereas it is high among NVHD patients; the rheumatic etiology does not seem to have any beneficial effects on the prevalence of CAD; gender, age, SAH, TACP, dyslipidemia and diabetes were identified as being strongly associated with the presence of CAD. It is possible to define the criteria that indicate the need for pre-surgical CA in heart valve replacements, so that the standard indication after the age of 40 years can be avoided. PMID- 18392401 TI - Case 2/2008: a 43 years old man with hallucination, nape rigidity, convulsion and auscultation of complete aortic valve dysfunction. PMID- 18392402 TI - Levosimendan use in several scenarios of acute heart failure. AB - In countries where it is available, early levosimendan infusion can be considered for patients who remain symptomatic with dyspnea at rest despite initial therapy, particularly those with a history of chronic heart failure or chronically treated with beta-blockers. Hypotensive patients or patients with active ischemia are not the best candidates for levosimendan administration and should have these problems addressed first. PMID- 18392403 TI - Neuromyelitis optica: brain abnormalities in a Brazilian cohort. AB - Neuromyelitis optica (NMO) is a demyelinating disease consisting of relapsing remitting optic neuritis and myelitis with a more severe course than Multiple Sclerosis. Recently, it has been shown that almost 50% of patients with NMO can have brain magnetic resonance imaging (MRI) abnormalities. We report on six Brazilian patients with NMO, fulfilling the 1999 Wingerchuck criteria for this disease, with abnormal brain MRI and discuss their clinical and radiological features. PMID- 18392404 TI - Demyelinating disease in patients with myasthenia gravis. AB - Myasthenia gravis (MG) is an autoimmune disease characterized by fluctuating muscle weakness, caused by impaired neuromuscular transmission. Patients with MG can present other autoimmune diseases in association, commonly hypo or hyperthyroidism. The association of MG to demyelinating disease is rare and has been described before. We report on three Brazilian patients with MG that presented distinct demyelinating diseases, two monophasic and one recurrent neuromyelitis optica, several years after the diagnosis of MG, and discuss their clinical courses. PMID- 18392405 TI - Beta interferons in clinically isolated syndromes: a meta-analysis. AB - Beta-interferon use in definite multiple sclerosis (MS) has been proven to modify clinical and magnetic resonance imaging outcome. We review and summarize the data of published double-blind, randomized clinical trials to assess, with a meta analysis the safety and efficacy of beta-interferon on the occurrence of relapses in patients with a first clinical event suggestive of MS. After two years of follow-up, interferon beta decreased the risk of conversion to clinically definite MS 0.51[0.39-0.65], and delayed the time to diagnosis up to 367 days. Side-effects were mild and self limited. Our findings support the efficacy of early treatment with beta-interferon in reducing conversion to clinically defined MS in patients with clinically isolated syndromes. PMID- 18392406 TI - Immunomodulator therapy migration in relapsing remitting multiple sclerosis: a study of 152 cases. AB - BACKGROUND: Since 1997, immunological modulators have been used for treatment of Relapsing Remitting Multiple Sclerosis (RRMS) in the Multiple Sclerosis Attendance and Treatment Center (CATEM) with significant alterations in this disease natural history. AIM: To add data on the experience of CATEM for the treatment of RRMS patients that had immunomodulators. METHOD: RRMS patients that received continuously immunomodulator drugs were evaluated on adherence, migration, withdrawal and progression rates. The patients were divided in three groups by the period of immunomodulators intake. RESULTS: There were registered in Group 1 withdrawal in 98 patients (25%) and adherence in 292 cases (74%); Group 2 interruption of therapy in 140 patients, 92 (31%) due to progression for PSMS, 14 (5%) for pregnancy, withdrawal in 34 (11%), adherence in 88%; Group 3 progression in 41 (26%), pregnancy in 3 (2%) withdrawal in 42 (27%) and adherence in 72%. The migration rate was about one third (31.57%) and the principal cause was therapeutic failure; the mean migrating time was 0.5-2.5 years in group 3. CONCLUSION: Immunomodulatory treatment for RRMS patients may have significant levels of failure and side effects; the adherence was compatible with the international literature. PMID- 18392407 TI - Cervical dystonia: clinical and therapeutic features in 85 patients. AB - We studied patients with cervical dystonia (CD) to determine clinical features and response to botulinum toxin A (BoNT/A). Patients were submitted to clinical, laboratory and neuroimaging evaluation. BoNT/A was injected locally in 81 patients using electromyographic guidance. Four patients who had had previous treatment were considered to be in remission. The average ages at onset of focal dystonia and segmental dystonia were greater than for generalized dystonia (p<0.0003). The severity of the abnormal head-neck movements were more severe among the patients with generalized dystonia (p<0.001). Pain in the cervical area was noted in 59 patients. It was not possible to determine the etiology of the disease in 62.3% of patients. Tardive dystonia was the most common secondary etiology. A major improvement in the motor symptoms of CD and pain was observed in patients following treatment with BoNT/A. The tardive dystonia subgroup did not respond to the treatment. Dysphagia was observed in 2.35% of the patients. PMID- 18392408 TI - Features associated with the development of non-motor manifestations in Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder, predominantly characterized by the presence of motor symptoms. However, the non motor manifestations (NMM) are a frequent complaint in the PD patients. There is a lack of information about the risk factors associated with the NMM in these patients. The aim of this study is to evaluate the prevalence of the more common NMM in a population of PD patients and to determine the features associated with its development. We studied 124 ambulatory PD patients. NMM were defined by the presence of neuropsychiatric manifestations, cognitive disorder, autonomic dysfunction or sleep related problems. In a multivariate analysis we found that the years of evolution of the PD and the presence of cognitive dysfunction are the risk factors for the neuropsychiatric and autonomic manifestations, whereas axial impairment is a risk factor for cognitive disorders and dyskinesias is for sleep related problems. In conclusion, this study shows that the features related to the PD progression appear as the main risk factors associated with NMM. PMID- 18392409 TI - Pain in Parkinson's disease: analysis of 50 cases in a clinic of movement disorders. AB - INTRODUCTION: Pain is a common symptom in Parkinson's disease (PD), and is often related to the illness itself. OBJECTIVE: To prospectively establish the occurrence of pain in PD patients. METHOD: This study was conducted within a population composed of 50 patients with PD to evaluate the presence of pain. RESULTS: Twenty-eight patients reported pain; comparing the group with pain and the group without pain, there were no differences related to the beginning of the illness and the motor symptoms of PD. However, many patients related an improvement of pain when antiparkinsonian therapy was initiated or adjusted. CONCLUSION: The use of techniques for analgesia and the adjustment of PD medication contribute to improve the manifestations of pain and the life quality of patients with PD. PMID- 18392410 TI - Stroke in patients with sickle cell disease: clinical and neurological aspects. AB - The aim of this study was to characterize a group of patients (n=8) with sickle cell disease (SCD) and ischemic stroke concerning the clinical, neurological, imaging and progressive aspects. Data were collected from records and completed with an interview of patients and their parents. In this study there were 8 patients with ages ranging from 10 to 23 years old; SCD diagnosis was given between one and two years of age with clinical features of fatigue and anemia. The stroke was ischemic in all individuals and the first cerebrovascular event occurred before 6 years of age; 3 patients had recurrence of stroke despite prophylactic blood transfusion therapy and both cerebral hemispheres were affected in 4 patients. Clinical and neurological current features observed were: acute pain crises, sialorrhea, mouth breathing, motor, and neuropsychological impairments resulting from cortical-subcortical structure lesions. PMID- 18392412 TI - Brazilian children performance on Rey's auditory verbal learning paradigm. AB - The Rey Auditory Verbal Learning paradigm is worldwide used in clinical and research settings. There is consensus about its psychometric robustessness and that its various scores provide relevant information about different aspects of memory and learning. However, there are only a few studies in Brazil employing this paradigm and none of them with children. This paper describes the performance of 119 Brazilian children in a version of Rey's paradigm. The correlations between scores showed the internal consistency of this version. Also, the pattern of results observed was very similar to that observed in foreign studies with adults and children. There was correlation between age in months and recall scores, showing that age affects the rhythm of learning. These results were discussed based on the information processing theory. PMID- 18392411 TI - Language brain dominance in patients with refractory temporal lobe epilepsy: a comparative study between functional magnetic resonance imaging and dichotic listening test. AB - PURPOSE: To identify brain dominance for language functions with DLT and correlate these results with those obtained from fMRI in patients suffering from intractable temporal lobe epilepsy. METHOD: This study reports on 13 patients who underwent pre-surgical epileptic evaluation between April and October 2004 at the Epilepsy Surgery Program, Hospital Sao Lucas, PUCRS. In DLT, dominance was assessed through a consonant-vowel task, whereas in fMRI patients performed a verb generation task. RESULTS: Our results identified a correlation between the fMRI lateralization index and the DLT ear predominance index and reply difference index (r=0.6, p=0.02; Pearson Correlation Coefficient), showing positive correlation between results obtained from fMRI and DLT. CONCLUSION: DLT was found to significantly correlate with fMRI. These findings indicate that DLT (a non invasive procedure) could be a useful tool to evaluate language brain dominance in pre-surgical epileptic patients as it is cheaper to perform than fMRI. PMID- 18392413 TI - Unusual dissemination patterns of low-grade astrocytomas in childhood. AB - CONTEXT: Low-grade astrocytomas are intracerebral lesions of relatively high frequency in the under-18 pediatric population. They often present indolent behaviour, and complete surgical resection is the choice treatment. In cases where the surgery is not possible, chemotherapy and radiotherapy may be used. Medical reports do not recommend examination of the spinal cord at diagnosis or during treatment, since the risk of dissemination of the lesion to the spine is minimal according to medical experience. We describe here four cases of children with low-grade astrocytoma with aggressive dissemination to the neuroaxis. PMID- 18392414 TI - Normative study of tympanic infrared thermometry: a non-invasive index of asymmetric cerebral activity. AB - Human and primate studies have demonstrated that performance of tasks that induce asymmetrical physiological activation of the cerebral hemispheres leads to a reduction of tympanic temperature (TT) ipsilateral to the most active hemisphere. It is possible that diseases that interfere in an asymmetrical fashion with the degree of cerebral activity cause similar TT changes. There are not, however, normative studies of the acceptable interaural difference in TT in normal subjects at rest. This study was done to establish normative values for interaural TT values measured by means of infrared tympanic thermometry in resting normal subjects not engaged in any specific task. TT values were measured in 47 normal volunteers (20 men and 27 women, aged 39.38+/-12.57 years old) at rest; mean interaural differences of TT were calculated. Mean right ear TT was 36.85+/-0.50 degrees C and mean left ear TT was 36.74+/-0.57 degrees C; these values are in agreement with those already reported in the literature. Mean interaural TT difference was 0.25 degrees C (SD 0.21 degrees C). These findings indicate that maximal normal values for interaural TT differences, with confidence levels of 99% and 95%, are, respectively, 0.88 and 0.67 degrees C. The value of interaural differences of TT as a marker of asymmetrical hemispheric activity in neurological patients will have to be established by additional studies. PMID- 18392415 TI - Clinical application of magnetic resonance in acute traumatic brain injury. AB - PURPOSE: To evaluate the clinical applications of magnetic resonance imaging (MRI) in patients with acute traumatic brain injury (TBI): to identify the type, quantity, severity; and improvement clinical-radiological correlation. METHOD: Assessment of 55 patients who were imaged using CT and MRI, 34 (61.8%) males and 21 (38.2%) females, with acute (0 to 5 days) and closed TBI. RESULTS: Statistical significant differences (McNemar test): ocurred fractures were detected by CT in 29.1% and by MRI in 3.6% of the patients; subdural hematoma by CT in 10.9% and MRI in 36.4 %; diffuse axonal injury (DAI) by CT in 1.8% and MRI in 50.9%; cortical contusions by CT in 9.1% and MRI in 41.8%; subarachnoid hemorrhage by CT in 18.2% and MRI in 41.8%. CONCLUSION: MRI was superior to the CT in the identification of DAI, subarachnoid hemorrhage, cortical contusions, and acute subdural hematoma; however it was inferior in diagnosing fractures. The detection of DAI was associated with the severity of acute TBI. PMID- 18392416 TI - Surgical treatment of primary intramedullary spinal cord tumors in adult patients. AB - BACKGROUND: Primary spinal cord intramedullary tumors are rare and present with insidious symptoms. Previous treatment protocols emphasized biopsy and radiation/chemotherapy but more aggressive protocols have emerged. OBJECTIVE: To report our experience. METHOD: Forty-eight patients were diagnosed with primary intramedullary tumors. The cervical cord was involved in 27% and thoracic in 42% of patients. Complete microsurgical removal was attempted whenever possible without added neurological morbidity. RESULTS: Complete resection was obtained in 33 (71%) patients. Neurological function remained stable or improved in 32 patients (66.7%). Ependymoma was the most frequent tumor (66.7%). CONCLUSION: Neurological outcome is superior in patients with subtle findings; aggressive microsurgical resection should be pursued with acceptable neurological outcomes. PMID- 18392417 TI - Diffusion-weighted MR images and pineoblastoma: diagnosis and follow-up. AB - Pineoblastomas are uncommon pineal tumors, which demonstrate rapid growing and poor prognosis. We report the case of a 43-year-old man with an enhancing pineal region mass, which showed restriction of the diffusion on diffusion-weighted (DW) MR images. The surgical biopsy defined the diagnosis of pineoblastoma and the therapy was initiated with radiation and chemotherapy. Three months later, the follow-up MR imaging showed areas suggestive of necrosis and the DW images demonstrate no significant areas of restricted diffusion. The differential diagnosis of pineal region masses that could show restriction of diffusion is discussed. PMID- 18392418 TI - Occurrence of severe and moderate traumatic brain injury in patients attended in a Brazilian Teaching Hospital: epidemiology and dosage of alcoholemy. AB - This study aimed at observing aspects of epidemiology in order to investigate the use of alcohol in patients older than 18 with severe and moderate traumatic brain injury, which were attended in the Clinics Hospital of the University of Uberlandia. Positive alcoholemy was found in 39.3% of the patients. Of the 33 positive exams alcoholemy was found higher than 60 mg/dL in 28 (84.6%). There was not significant relation between alcoholemy levels and trauma severity. The major prevalence occurred on Saturdays nights. The most frequent types of external causes were transportation accidents (64.74) followed by accidental falls (17.27%) and physical aggression (16.55%). 93.9% of the patients with positive alcoholemy were men aged 20-29. 24.2% of the ones with positive alcoholemy died yet no significant difference was found in the study of the ones with negative alcoholemy (n=51) (p=0.93); RR= 0.9; IC95%=0.40-2.08. PMID- 18392419 TI - Occipital neuroma triggered cluster headache responding to greater occipital nerve blockade. PMID- 18392421 TI - Limb-girdle muscular dystrophy type 2B mimicking polymyositis. PMID- 18392420 TI - Episodic migraine associated with postural orthostatic tachycardia syndrome and vasovagal syncope: migraine triggers neuromediated syncope. PMID- 18392422 TI - Focal facial spasms associated with benign parotid hypertrophy. PMID- 18392423 TI - Primary diffuse leptomeningeal gliomatosis. PMID- 18392424 TI - Infantile gliosarcoma. PMID- 18392425 TI - Median (third) occipital condyle causing atlantoaxial instability and myelopathy. PMID- 18392426 TI - Penetrating screwdriver wound to the head. PMID- 18392427 TI - Ice pack test in the diagnosis of myasthenia gravis. PMID- 18392429 TI - Hemiparesis and lateralized cortical lesions associated to severe hypoglycemia. PMID- 18392428 TI - Intrasellar internal carotid aneurysm coexisting with GH-secreting pituitary adenoma in an acromegalic patient. PMID- 18392430 TI - Brain abscess following ischemic stroke with secondary hemorrhage. PMID- 18392431 TI - Chorea due to gabapentin monotherapy in a not encephalopatic patient. PMID- 18392432 TI - Optic neuritis due to solvent abuse. PMID- 18392433 TI - Migrating intraventricular cysticercosis: magnetic resonance imaging findings. PMID- 18392434 TI - Treatment of cerebral cysticercosis with albendazole in elevated dosages. PMID- 18392435 TI - A brief historical note on the classification of nerve fibers. AB - This is a brief review of the literature focused on the articles that formed the basis for the classification of the nerve fibers. Mention is also made to the origin of the nomenclature of the different motoneurons (alpha, beta and gamma). PMID- 18392436 TI - Devic's neuromyelitis optica: a critical review. AB - Devic's neuromyelitis optica (NMO) is an idiopathic inflammatory demyelinating and necrotizing disease characterized by predominant involvement of the optic nerves and spinal cord. In Asian countries relapsing NMO has been known as opticospinal multiple sclerosis. It has long been debated if NMO is a variant of multiple sclerosis (MS) or a distinct disease. Recent studies have shown that NMO has more frequently a relapsing course, and results from attack to aquaporin-4 which is the dominant water channel in the central nervous system, located in foot processes of the astrocytes. Distinctive pathological features of NMO include perivascular deposition of IgG and complement in the perivascular space, granulocyte and eosinophil infiltrates and hyalinization of the vascular walls. These features distinguish NMO from other demyelinating diseases such as MS and acute demyelinating encephalomyelopathy. An IgG-antibody that binds to aquaporin 4, named NMO-IgG has high sensitivity and specificity. Magnetic resonance imaging (MRI) studies have revealed that more frequently there is a long spinal cord lesion that extends through three or more vertebral segments in length. Brain MRI lesions atypical for MS are found in the majority of cases. Treatment in the acute phase includes intravenous steroids and plasma exchange therapy. Immunosupressive agents are recommended for prophylaxis of relapses. PMID- 18392441 TI - Cognitive impairment in Parkinson's disease. PMID- 18392442 TI - Glossopharyngeal neuralgia with syncope as a sign of neck cancer recurrence. PMID- 18392447 TI - Comparison of scientists of the Brazilian Academy of Sciences and of the National Academy of Sciences of the USA on the basis of the h-index. AB - A new scientometric indicator, the h-index, has been recently proposed (Hirsch JE. Proc Natl Acad Sci 2005; 102: 16569-16572). The index avoids some shortcomings of the calculation of the total number of citations as a parameter to evaluate scientific performance. Although it has become known only recently, it has had widespread acceptance. A comparison of the average h-index of members of the Brazilian Academy of Sciences (BAS) and of the National Academy of Sciences of the USA (NAS-USA) was carried out for 10 different areas of science. Although, as expected, the comparison was unfavorable to the members of the BAS, the imbalance was distinct in different areas. Since these two academies represent, to a significant extent, the science of top quality produced in each country, the comparison allows the identification of the areas in Brazil that are closer to the international stakeholders of scientific excellence. The areas of Physics and Mathematics stand out in this context. The heterogeneity of the h index in the different areas, estimated by the median dispersion of the index, is significantly higher in the BAS than in the NAS-USA. No elements have been collected in the present study to provide an explanation for this fact. PMID- 18392448 TI - Serotonergic modulation of face-emotion recognition. AB - Facial expressions of basic emotions have been widely used to investigate the neural substrates of emotion processing, but little is known about the exact meaning of subjective changes provoked by perceiving facial expressions. Our assumption was that fearful faces would be related to the processing of potential threats, whereas angry faces would be related to the processing of proximal threats. Experimental studies have suggested that serotonin modulates the brain processes underlying defensive responses to environmental threats, facilitating risk assessment behavior elicited by potential threats and inhibiting fight or flight responses to proximal threats. In order to test these predictions about the relationship between fearful and angry faces and defensive behaviors, we carried out a review of the literature about the effects of pharmacological probes that affect 5-HT-mediated neurotransmission on the perception of emotional faces. The hypothesis that angry faces would be processed as a proximal threat and that, as a consequence, their recognition would be impaired by an increase in 5-HT function was not supported by the results reviewed. In contrast, most of the studies that evaluated the behavioral effects of serotonin challenges showed that increased 5-HT neurotransmission facilitates the recognition of fearful faces, whereas its decrease impairs the same performance. These results agree with the hypothesis that fearful faces are processed as potential threats and that 5-HT enhances this brain processing. PMID- 18392449 TI - Molecular analysis of the bovine coronavirus S1 gene by direct sequencing of diarrheic fecal specimens. AB - Bovine coronavirus (BCoV) causes severe diarrhea in newborn calves, is associated with winter dysentery in adult cattle and respiratory infections in calves and feedlot cattle. The BCoV S protein plays a fundamental role in viral attachment and entry into the host cell, and is cleaved into two subunits termed S1 (amino terminal) and S2 (carboxy terminal). The present study describes a strategy for the sequencing of the BCoV S1 gene directly from fecal diarrheic specimens that were previously identified as BCoV positive by RT-PCR assay for N gene detection. A consensus sequence of 2681 nucleotides was obtained through direct sequencing of seven overlapping PCR fragments of the S gene. The samples did not undergo cell culture passage prior to PCR amplification and sequencing. The structural analysis was based on the genomic differences between Brazilian strains and other known BCoV from different geographical regions. The phylogenetic analysis of the entire S1 gene showed that the BCoV Brazilian strains were more distant from the Mebus strain (97.8% identity for nucleotides and 96.8% identity for amino acids) and more similar to the BCoV-ENT strain (98.7% for nucleotides and 98.7% for amino acids). Based on the phylogenetic analysis of the hypervariable region of the S1 subunit, these strains clustered with the American (BCoV-ENT, 182NS) and Canadian (BCQ20, BCQ2070, BCQ9, BCQ571, BCQ1523) calf diarrhea and the Canadian winter dysentery (BCQ7373, BCQ2590) strains, but clustered on a separate branch of the Korean and respiratory BCoV strains. The BCoV strains of the present study were not clustered in the same branch of previously published Brazilian strains (AY606193, AY606194). These data agree with the genealogical construction and suggest that at least two different BCoV strains are circulating in Brazil. PMID- 18392450 TI - Transient high-level expression of beta-galactosidase after transfection of fibroblasts from GM1 gangliosidosis patients with plasmid DNA. AB - GM1 gangliosidosis is an autosomal recessive disorder caused by the deficiency of lysosomal acid hydrolase beta-galactosidase (beta-Gal). It is one of the most frequent lysosomal storage disorders in Brazil, with an estimated frequency of 1:17,000. The enzyme is secreted and can be captured by deficient cells and targeted to the lysosomes. There is no effective treatment for GM1 gangliosidosis. To determine the efficiency of an expression vector for correcting the genetic defect of GM1 gangliosidosis, we tested transfer of the beta-Gal gene (Glb1) to fibroblasts in culture using liposomes. Beta-Gal cDNA was cloned into the expression vectors pSCTOP and pREP9. Transfection was performed using 4 microL lipofectamine 2000 and 1.5-2.0 microg DNA. Cells (2 x 10(5)/well) were harvested 24 h, 48 h, and 7 days after transfection. Enzyme specific activity was measured in cell lysate and supernatant by fluorometric assay. Twenty-four hours after transfection, treated cells showed a higher enzyme specific activity (pREP9-beta-Gal: 621.5 +/- 323.0, pSCTOP-beta-Gal: 714.5 +/- 349.5, pREP9-beta-Gal + pSCTOP-beta-Gal: 1859.0 +/- 182.4, and pREP9-ss-Gal + pTRACER: 979.5 +/- 254.9 nmol x h-1 x mg-1 protein) compared to untreated cells (18.0 +/- 3.1 for cell and 32.2 +/- 22.2 nmol x h-1 x mg-1 protein for supernatant). However, cells maintained in culture for 7 days showed values similar to those of untreated patients. In the present study, we were able to transfect primary patients' skin fibroblasts in culture using a non-viral vector which overexpresses the beta-Gal gene for 24 h. This is the first attempt to correct fibroblasts from patients with GM1 gangliosidosis by gene therapy using a non-viral vector. PMID- 18392451 TI - Different responses of the GlnB and GlnZ proteins upon in vitro uridylylation by the Azospirillum brasilense GlnD protein. AB - Azospirillum brasilense is a diazotroph found in association with important agricultural crops. In this organism, the regulation of nitrogen fixation by ammonium ions involves several proteins including the uridylyltransferase/uridylyl-removing enzyme, GlnD, which reversibly uridylylates the two PII proteins, GlnB and GlnZ, in response to the concentration of ammonium ions. In the present study, the uridylylation/deuridylylation cycle of A. brasilense GlnB and GlnZ proteins by GlnD was reconstituted in vitro using the purified proteins. The uridylylation assay was analyzed using non-denaturing polyacrylamide gel electrophoresis and fluorescent protein detection. Our results show that the purified A. brasilense GlnB and GlnZ proteins were uridylylated by the purified A. brasilense GlnD protein in a process dependent on ATP and 2 oxoglutarate. The dependence on ATP for uridylylation was similar for both proteins. On the other hand, at micromolar concentration of 2-oxoglutarate (up to 100 microM), GlnB uridylylation was almost twice that of GlnZ, an effect that was not observed at higher concentrations of 2-oxoglutarate (up to 10 mM). Glutamine inhibited uridylylation and stimulated deuridylylation of both GlnB and GlnZ. However, glutamine seemed to inhibit GlnZ uridylylation more efficiently. Our results suggest that the differences in the uridylylation pattern of GlnB and GlnZ might be important for fine-tuning of the signaling pathway of cellular nitrogen status in A. brasilense. PMID- 18392452 TI - Intrinsic bent DNA sites in the chromosomal replication origin of Xylella fastidiosa 9a5c. AB - The features of the nucleotide sequences in both replication and promoter regions have been investigated in many organisms. Intrinsically bent DNA sites associated with transcription have been described in several prokaryotic organisms. The aim of the present study was to investigate intrinsic bent DNA sites in the segment that holds the chromosomal replication origin, oriC, of Xylella fastidiosa 9a5c. Electrophoretic behavior analyses, as well as in silico analyses of both the 2-D projection and helical parameters, were performed. The chromosomal segment analyzed contains the initial sequence of the rpmH gene, an intergenic region, the dnaA gene, the oriC sequence, and the 5' partial sequence of the dnaN gene. The analysis revealed fragments with reduced electrophoretic mobility, which indicates the presence of curved DNA segments. The analysis of the helical parameter ENDS ratio revealed three bent DNA sites (b1, b2, and b3) located in the rpmH-dnaA intergenic region, the dnaA gene, and the oriC 5' end, respectively. The chromosomal segment of X. fastidiosa analyzed here is rich in phased AT tracts and in CAnT motifs. The 2-D projection indicated a segment whose structure was determined by the cumulative effect of all bent DNA sites. Further, the in silico analysis of the three different bacterial oriC sequences indicated similar negative roll and twist >34.00 degrees values. The DnaA box sequences, and other motifs in them, may be associated with the intrinsic DNA curvature. PMID- 18392453 TI - Paullinia cupana Mart var. sorbilis, guarana, reduces cell proliferation and increases apoptosis of B16/F10 melanoma lung metastases in mice. AB - We showed that guarana (Paullinia cupana Mart var. sorbilis) had a chemopreventive effect on mouse hepatocarcinogenesis and reduced diethylnitrosamine-induced DNA damage. In the present experiment, we evaluated the effects of guarana in an experimental metastasis model. Cultured B16/F10 melanoma cells (5 x 10(5) cells/animal) were injected into the tail vein of mice on the 7th day of guarana treatment (2.0 mg P. cupana/g body weight, per gavage) and the animals were treated with guarana daily up to 14 days until euthanasia (total treatment time: 21 days). Lung sections were obtained for morphometric analysis, apoptotic bodies were counted to calculate the apoptotic index and proliferating cell nuclear antigen-positive cells were counted to determine the proliferation index. Guarana-treated (GUA) animals presented a 68.6% reduction in tumor burden area compared to control (CO) animals which were not treated with guarana (CO: 0.84 +/- 0.26, N = 6; GUA: 0.27 +/- 0.24, N = 6; P = 0.0043), a 57.9% reduction in tumor proliferation index (CO: 23.75 +/- 20.54, N = 6; GUA: 9.99 +/- 3.93, N = 6; P = 0.026) and a 4.85-fold increase in apoptotic index (CO: 66.95 +/- 22.95, N = 6; GUA: 324.37 +/- 266.74 AB/mm(2), N = 6; P = 0.0152). In this mouse model, guarana treatment decreased proliferation and increased apoptosis of tumor cells, consequently reducing the tumor burden area. We are currently investigating the molecular pathways of the effects of guarana in cultured melanoma cells, regarding principally the cell cycle inhibitors and cyclins. PMID- 18392455 TI - Defensive responses to threat scenarios in Brazilians reproduce the pattern of Hawaiian Americans and non-human mammals. AB - A former study with scenarios conducted in Hawaii has suggested that humans share with non-human mammals the same basic defensive strategies - risk assessment, freezing, defensive threat, defensive attack, and flight. The selection of the most adaptive strategy is strongly influenced by features of the threat stimulus magnitude, escapability, distance, ambiguity, and availability of a hiding place. Aiming at verifying if these strategies would be consistent in a different culture, 12 defensive scenarios were translated into Portuguese and adapted to the Brazilian culture. The sample consisted of male and female undergraduate students divided into two groups: 76 students, who evaluated the five dimensions of each scenario and 248 medical students, who chose the most likely response for each scenario. In agreement with the findings from studies of non-human mammal species, the scenarios were able to elicit different defensive behavioral responses, depending on features of the threat. "Flight" was chosen as the most likely response in scenarios evaluated as an unambiguous and intense threat, but with an available route of escape, whereas "attack" was chosen in an unambiguous, intense and close dangerous situation without an escape route. Less urgent behaviors, such as "check out", were chosen in scenarios evaluated as less intense, more distant and more ambiguous. Moreover, the results from the Brazilian sample were similar to the results obtained in the original study with Hawaiian students. These data suggest that a basic repertoire of defensive strategies is conserved along the mammalian evolution because they share similar functional benefits in maintaining fitness. PMID- 18392454 TI - Locally produced mucosal IgG in chickens immunized with conventional vaccines for Newcastle disease virus. AB - Newcastle disease virus (NDV) is the causative agent of an economically important disease, which affects all species of birds worldwide. Current vaccination programs for NDV include the use of either low-virulent live-virus vaccines or inactivated vaccines to induce protective immunity while producing minimal adverse effects in birds. In order to further characterize the immune response elicited by live virus and inactivated NDV conventional vaccines in chickens, we evaluated the presence of specific antibodies in different secretions and in tissue culture supernatants of immunized birds. To this end, we analyzed all the samples by ELISA, using an indirect assay set up in the laboratory. Specific anti NDV IgG antibodies were detected in tracheal and cloacal swabs and tracheal and intestinal washes of immunized animals. We also found specific anti-NDV IgG antibodies in tracheal and intestinal tissue culture supernatants, indicating that the IgG found in swabs and washes was not transudated from serum or, at least, was not all transudated from serum. Knowledge about the mechanisms involved in the immune response of chickens to different NDV vaccines should increase our understanding of the mucosal response against the virus and, eventually, provide new useful information for the development and evaluation of synthetic vaccines. PMID- 18392456 TI - Expression of neuronal nitric oxide synthase in the hippocampal formation in affective disorders. AB - Hippocampal output is increased in affective disorders and is mediated by increased glutamatergic input via N-methyl-D-aspartate (NMDA) receptor and moderated by antidepressant treatment. Activation of NMDA receptors by glutamate evokes the release of nitric oxide (NO) by the activation of neuronal nitric oxide synthase (nNOS). The human hippocampus contains a high density of NMDA receptors and nNOS-expressing neurons suggesting the existence of an NMDA-NO transduction pathway which can be involved in the pathogenesis of affective disorders. We tested the hypothesis that nNOS expression is increased in the human hippocampus from affectively ill patients. Immunocytochemistry was used to demonstrate nNOS-expressing neurons in sections obtained from the Stanley Consortium postmortem brain collection from patients with major depression (MD, N = 15), bipolar disorder (BD, N = 15), and schizophrenia (N = 15) and from controls (N = 15). nNOS-immunoreactive (nNOS-IR) and Nissl-stained neurons were counted in entorhinal cortex, hippocampal CA1, CA2, CA3, and CA4 subfields, and subiculum. The numbers of Nissl-stained neurons were very similar in different diagnostic groups and correlated significantly with the number of nNOS-IR neurons. Both the MD and the BD groups had greater number of nNOS-IR neurons/400 microm(2) in CA1 (mean +/- SEM: MD = 9.2 +/- 0.6 and BD = 8.4 +/- 0.6) and subiculum (BD = 6.7 +/- 0.4) when compared to control group (6.6 +/- 0.5) and this was significantly more marked in samples from the right hemisphere. These changes were specific to affective disorders since no changes were seen in the schizophrenic group (6.7 +/- 0.8). The results support the current view of the NMDA-NO pathway as a target for the pathophysiology of affective disorders and antidepressant drug development. PMID- 18392459 TI - Histological diagnosis of lung cancer micrometastases. PMID- 18392460 TI - Detection of micrometastases in pN0 non-small cell lung cancer: an alternative method combining tissue microarray and immunohistochemistry. AB - OBJECTIVE: To present an alternative method of detecting micrometastases in lymph nodes previously testing negative for non-small cell lung cancer (NSCLC) by routine hematoxylin-eosin staining. METHODS: A total of 77 hilar and mediastinal lymph nodes resected from 18 patients with NSCLC were investigated for the presence of micrometastases using a combination of microarray analysis and immunohistochemistry. RESULTS: Micrometastases were detected by identifying cytokeratin- and chromogranin-positive cells in lymph node microarrays. Of the 18 patients initially staged as pN0 through routine hematoxylin-eosin staining, 9 (50%) were restaged as N1, and the prognoses were re-evaluated in terms of histological and clinical parameters. The comparison of the survival curves revealed that survival was higher in the patients without micrometastases than in those with micrometastases. In addition, in the multivariate analysis adjusted for age, gender, histological type, and restaging, the presence of micrometastases proved to be an independent predictor of survival. Among patients who had been previously staged as pN0, the risk of death was found to be 7-times greater for those later diagnosed with micrometastases than for those in whom no micrometastases were identified. CONCLUSION: The combination of microarray analysis and immunohistochemistry might represent a low-cost and less time consuming alternative for identifying occult micrometastases and predicting prognoses in surgically resected patients with pN0 NSCLC. Larger randomized, prospective studies are needed in order to determine the accuracy of this method. PMID- 18392461 TI - The incidence of residual pneumothorax after video-assisted sympathectomy with and without pleural drainage and its effect on postoperative pain. AB - OBJECTIVE: To determine the incidence of residual pneumothorax after video assisted thoracic sympathectomy, with and without postoperative pleural drainage, and to evaluate the possible influence of this type of pneumothorax on postoperative pain within the first 28 postoperative days. METHODS: All patients presenting symptoms consistent with primary palmoplantar hyperhidrosis and treated at the Thoracic Surgery Outpatient Clinic of the State Hospital of Sumare between July and December of 2006 were included. All were submitted to sympathectomy up to the third ganglion using video-assisted thoracoscopy and were randomized to receive or not receive postoperative pleural drainage for 3 h. Chest X-rays and low-dose computed tomography scans of the chest were performed on the first postoperative day in order to determine the incidence of residual pneumothorax. At different time points up to postoperative day 28, patient pain was assessed using a visual numeric scale and by measuring the quantity of opioid analgesics required. RESULTS: This study comprised 56 patients, 27 submitted to bilateral pleural drainage and 29 not submitted to drainage. There was no statistical difference between the two groups in terms of the incidence of post sympathectomy residual pneumothorax. Residual pneumothorax diagnosed through any of the methods did not influence pain within the first 28 postoperative days. CONCLUSION: Performing closed pleural drainage for 3 h immediately after video assisted thoracic sympathectomy did not affect lung re-expansion or the incidence of residual pneumothorax. When residual pneumothorax was present, it did not affect pain within the first 28 postoperative days. PMID- 18392462 TI - The Brazilian Portuguese version of the London Chest Activity of Daily Living scale for use in patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: To translate the London Chest Activity of Daily Living (LCADL) scale into Portuguese and to determine whether this version is reproducible in Brazilian patients with severe chronic obstructive pulmonary disease (COPD). METHODS: The LCADL scale was translated into Portuguese and then back-translated into English. This pilot Brazilian Portuguese version was administered to 8 patients with COPD, and possible text-related problems were investigated. The principal problems were discussed with the authors of the original scale, and a final translated version was arrived at. At the study outset, two observers administered this final version (twice in one day) to 31 patients with COPD. One of those observers again administered the scale to the same patients 15-20 days later. At baseline, the patients were submitted to pulmonary function testing and to the six-minute walk test (6MWT). RESULTS: The Brazilian Portuguese version of the LCADL scale demonstrated excellent reproducibility in the total score and in most of the questions, with an inter-rater Cronbach's alpha coefficient of 0.97 (95% CI: 0.89-0.97; p < 0.01) and an intra-rater Cronbach's alpha coefficient of 0.96 (95% CI: 0.83-0.96; p < 0.01). The total score presented a negative correlation with forced expiratory volume in one second in liters (r = -0.49; p < 0.05) and with distance covered on the 6MWT (r = -0.56; p < 0.05). CONCLUSION: The Brazilian Portuguese version of the LCADL scale is a reliable, reproducible, and valid instrument for evaluating dyspnea during activities of daily living in patients with severe COPD. PMID- 18392464 TI - Noncompliance with tuberculosis treatment involving self administration of treatment or the directly observed therapy, short-course strategy in a tuberculosis control program in the city of Carapicuiba, Brazil. AB - OBJECTIVE: To determine treatment noncompliance rates among patients participating in a municipal tuberculosis control program and to identify the variables related to noncompliance depending on the type of treatment strategy used. METHODS: A longitudinal non-concurrent cohort study was carried out involving two cohorts of patients participating in the Tuberculosis Control Program of the city of Carapicuiba, Brazil. The first cohort comprised 173 patients with tuberculosis treated from January 1, 2003 to December 31, 2003 using self administration of treatment, and the second comprised 187 patients with tuberculosis treated from July 1, 2004 to June 30, 2005 using the directly observed therapy, short-course strategy. RESULTS: Noncompliance rates decreased from 13.3% (self administration of treatment) to 5.9% (directly observed therapy, short-course), a significant difference (p < 0.05). For the self administration of treatment strategy, the variables significantly associated with treatment noncompliance were as follows: being an unregistered worker (relative risk [RR] = 3.06); retreatment (RR = 2.73); alcoholism (RR = 3.10); and no investigation of contacts (RR = 8.94). For the directly observed therapy, short-course strategy, no variables were significantly associated with noncompliance. CONCLUSION: The directly observed therapy, short-course strategy decreased noncompliance rates and produced better treatment outcomes, even when the risk factors for noncompliance were the same. PMID- 18392463 TI - Sputum examination in the clinical management of community-acquired pneumonia. AB - OBJECTIVE: To evaluate the frequency of the use of sputum examination in the clinical management of community-acquired pneumonia (CAP) in a general hospital and to determine whether its use has an impact on mortality. METHODS: The medical records of CAP patients treated as inpatients between May and November of 2004 at the Nossa Senhora da Conceicao Hospital, located in Porto Alegre, Brazil, were reviewed regarding the following aspects: age; gender; severity of pneumonia (Fine score); presence of sputum; sputum bacteriology; treatment history; change in treatment; and mortality. RESULTS: A total of 274 CAP patients (134 males and 140 females) were evaluated. Using the Fine score to quantify severity, we classified 79 (28.8%) of those 274 patients as class II, 45 (16.4%) as class III, 97 (35.4%) as class IV, and 53 (19.3%) as class V. Sputum examination was carried out in 92 patients (33.6%). A valid sample was obtained in 37 cases (13.5%), and an etiological diagnosis was obtained in 26 (9.5%), resulting in a change of treatment in only 9 cases (3.3%). Overall mortality was 18.6%. Advanced age (above 65), CAP severity, and dry cough were associated with an increase in the mortality rate. Sputum examination did not alter any clinical outcome or have any influence on mortality. CONCLUSION: Sputum examination was used in a minority of patients and was not associated with any noticeable benefit in the clinical management of patients with CAP treated in a hospital setting. PMID- 18392465 TI - Adapting the Bird Mark 7 to deliver noninvasive continuous positive airway pressure: a bench study. AB - OBJECTIVE: To test the efficiency of the Bird Mark 7 ventilator adapted to deliver continuous positive airway pressure (CPAP) in noninvasive positive pressure ventilation. METHODS: This was an experimental study using a mechanical model of the respiratory system. A Bird Mark 7 ventilator was supplied with 400 and 500 kPa and tested at CPAP of 5, 10 and 15 cmH2O. The following variables were analyzed: difference between the preset CPAP and the CPAP actually attained CPAP (trueCPAP); area of airway pressure at the CPAP level employed (AREA CPAP); and tidal volume generated. RESULTS: Adapting the Bird Mark 7 to offer CPAP achieved the expected tidal volume in all situations of inspiratory effort (normal or high), ventilator pressure supply (400 or 500 kPa) and CPAP value (5, 10 or 15 cmH2O). At a CPAP of 5 or 10 cmH2O, the trueCPAP was near the preset level, and the AREA CPAP was near zero. However, at a CPAP of 15 cmH2O, the value remained below the preset, and the AREA CPAP was high. CONCLUSION: The efficiency of Bird Mark 7 adaptation in offering CPAP was satisfactory at 5 and 10 cmH2O but insufficient at 15 cmH2O. If adapted as described in our study, the Bird Mark 7 might be an option for offering CPAP up to 10 cmH2O in areas where little or no equipment is available. PMID- 18392466 TI - Diagnosis of circadian rhythm sleep disorders. AB - Insomnia and excessive sleepiness are common in the investigation of sleep disordered breathing. Circadian rhythm sleep disorders are perhaps the most often overlooked conditions in the differential diagnosis of these symptoms. Circadian rhythm sleep disorders manifest as misalignment between the sleep period and the physical/social 24-h environmental cycle. The two most prevalent circadian rhythm sleep disorders are delayed sleep phase (common in adolescents) and advanced sleep phase (common in the elderly), situations in which the sleep period is displaced to a later or earlier time, respectively. It is important to keep these two disorders in mind, since they can be confused with insomnia and excessive sleepiness. However, there are nine possible diagnoses, and all nine are of clinical interest. Since light is the principal cue used in synchronizing the biological clock, blind individuals and night-shift/swing-shift workers are more prone to develop circadian rhythm sleep disorders. In this article, the new international classification of circadian rhythm sleep disorders is reviewed. PMID- 18392467 TI - Chronic eosinophilic pneumonia secondary to long-term use of nitrofurantoin: high resolution computed tomography findings. AB - The authors report the case of a female patient who developed chronic eosinophilic pneumonia secondary to long-term use of nitrofurantoin for prophylaxis of recurrent urinary tract infections due to urethral stenosis. On high-resolution computed tomography scans, the pulmonary reaction to nitrofurantoin most commonly manifests as an interstitial-alveolar pattern in both lung bases. However, in this case, the alterations were most pronounced in the periphery of the upper lobes. In itself, this tomographic profile is strongly indicative of chronic eosinophilic pneumonia. The patient had previously been submitted to an open lung biopsy. The diagnosis of chronic eosinophilic pneumonia was confirmed through a review of the biopsy. PMID- 18392468 TI - Giant cell tumor of the rib occupying the entire hemithorax. AB - The authors report the case of a 28-year-old female patient with a giant cell tumor originating from the rib. The tumor, measuring 25 x 17 cm, occupied the entire hemithorax and caused atelectasis of the left lung. This tumor was a benign mesenchymal neoplasm, which rarely affects the ribs. A thoracotomy involving en bloc resection of the chest wall and tumor was performed. Despite the large dimensions of the tumor, complete resection was possible, and lung function was restored. PMID- 18392469 TI - [Does the strict blood glucose control worsen cardiovascular risk in type 2 diabetes ?]. PMID- 18392470 TI - [Ill-defined causes of death and unattended deaths, Brazil, 2003]. PMID- 18392471 TI - [Is postural reeducation a useful tool for treatment of female stress urinary incontinence ?]. PMID- 18392472 TI - [Which is the importance of the budgetary analysis of projects in clinical research?]. PMID- 18392473 TI - [Resuscitation intra uterine in childbirth labor: Is there feasibility currently in Brazil? ]. PMID- 18392474 TI - [Thromboembolism and breast cancer: when should it be indicated the prophylactic medication? ]. PMID- 18392475 TI - [Prenatal care: part II]. PMID- 18392476 TI - [I stopped smoking!]. PMID- 18392477 TI - [Acute bronchiolitis, an updated review]. PMID- 18392479 TI - [Frantz's tumor (solid cystic tumor of the pancreas). ]. PMID- 18392480 TI - [Evaluation of the cumulative effect of meglumine antimoniate on the progeny of swiss mice: biologic assay]. AB - OBJECTIVES: Evaluate the effect of Meglumine Antimoniate on maternal-fetal transference in F1 generations (offspring of dams exposed to the drug), and embryotoxicity in F2 generations. METHODS: Female Swiss mice were treated with daily s.c. injection of Meglumine Antimoniate (100mgSb v/kg bw/day) from day 7 until day 12 of pregnancy. The control group received only the vehicle (distilled water). After birth of offspring (F1 generation), 59 females were examined daily for determination of the estral cycle. When the cycle estrus was determined, males were mated with 18 females of the same lineage. On day 18 of pregnancy, females were euthanasied in a chamber of CO2 and after incision of the abdomen, the uterus was exposed. Then, resorptions as well as living and dead fetuses were evaluated, also the number of embryo/fetal implantation sites. Fetuses and their placenta were weighted to calculate the placental index. Three placentas of each litter were separated for microscopic analysis. RESULTS: Administration of the Meglumine Antimoniate did not interfere in the estral cycle of the treated group, since it did not alter the precoital interval and fertility index. Placenta alterations were not observed in the F2 generations. CONCLUSION: Meglumine Antimoniate did not interfere in the reproductive performance, after chronic exposition of dams. Data suggest that there is a gradual elimination of Meglumine Antimoniate by the maternal organism without damaging the future offspring. PMID- 18392481 TI - [Impact of global postural reeducation for treatment of female stress urinary incontinence]. AB - OBJECTIVE: To evaluate the effect of global postural reeducation (GPR) on stress urinary incontinence (SUI) and quality of life in SUI female patients METHODS: The study design was a prospective non-randomized clinical trial. Twenty-six patients with symptoms of SUI were selected from the Urogynecology Outpatient Clinics of the State University of Campinas (Unicamp), state of Sao Paulo, Brazil. Age ranged from 23 to 72 years old (mean 50.8). All women were submitted to anamnesis, physical exam, postural evaluation and urodynamic testing. Patients were treated by the GPR in individual 50 minute sessions weekly for three months and twice a month for the next three months. All patients were re-evaluated at the end of treatment and six months later by means of General Impression of Improvement, Incontinence Impact, General Perception of Health, Functional Evaluation of the Pelvic Floor, Number of Leaking Episodes and Pad Use. RESULTS: At the end of treatment 4 (16%) of the patients were cured, 18 (72%) had improved significantly and 3 (12%) failed. At 6 months, 6 (24%) were cured, 16 (64%) improved and 3 (12%) failed (p<0.001). Quality of Life questionnaires presented significant improvement (p<0.05) in all domains, with emphasis on General Perception of Health, Incontinence Impact and number of leaking episodes. The Functional Evaluation of the Pelvic Floor and Pad Use also presented significant (p<0.001) improvement. CONCLUSION: These results may demonstrate that GPR is an efficient alternative for treatment of stress urinary incontinence. PMID- 18392482 TI - [Ill-defined causes of death and unattended deaths, Brazil, 2003]. AB - BACKGROUND: We studied the distribution of deaths from ill-defined causes that occurred in Brazil during 2003, from which was identified the proportion of unattended deaths. METHODS: Data were obtained from the Mortality Information System, coordinated by the Ministry of Health. Causes of death included in "Chapter XVIII - Symptoms, signs and abnormal clinical and laboratory findings, not classified elsewhere" of the International Statistical Classification of Diseases and Related Health Problems, tenth revision, were considered ill defined, among which the category R98 identified "unattended deaths". RESULTS: In Brazil during 2003 the underlying causes of 13.3% of deaths were included in the Chapter of ill-defined causes, and the highest proportions of these deaths occurred in the Northeast and North Regions. Considering the total deaths from ill-defined causes, 53 % correspond to unattended deaths. This proportion increased to over 70% in the states of Maranhao, Piaui, Rio Grande do Norte, Pernambuco, Bahia, Paraiba and Alagoas. CONCLUSION: Due to the decentralized structure of data collection in the country, we believe that the municipalities bear the major responsibility, followed by the states, for upgrading the quality of mortality statistics. PMID- 18392483 TI - [Knowledge of pregnant adolescents about reproductive anatomy and physiology in a municipality of Southern Brazil]. AB - OBJECTIVES: To study knowledge of some aspects of the female reproductive anatomy and physiology and their association with socio-demographic and reproductive "choices" of pregnant adolescents. METHODS: A cross-sectional study was performed with 200 first time pregnant adolescents who attended a public women's health clinic in the municipality of Indaiatuba, Sao Paulo, Brazil. During their first prenatal care visit, face-to-face interviews were conducted using a structured questionnaire and a three-dimensional handmade female model. Bivariate data analyses were performed using Pearson's Chi-square or Fisher's Exact test. Data were also analyzed using multivariate logistic regression models to test for associations of indicators of knowledge of female reproductive anatomy, physiology of female reproductive organs and physiology of reproduction with socio-demographic characteristics and reproductive "choices". RESULTS: The majority had little knowledge of anatomy (55.5%), with external organs more easily identified and placed than the internal; of physiology of reproductive organs (61.0%), and of physiology of reproduction (76.5%). Associations were found between knowledge and age of partner, couple difference of age, maintenance of the relationship with partner after pregnancy, religious affiliation, and level of education. No association was found between indicators of knowledge with use of contraceptives at first intercourse and with intention of having the baby at that time. CONCLUSION: This study addressed the complexity of the relationship between knowledge of reproductive anatomy and physiology and the theme of adolescent pregnancy, and emphasized the need for more contextualized approaches of programmatic contents on sexual education, in view of the intention to reduce early pregnancy. PMID- 18392484 TI - [Characterization of chronic pain and analgesic approaches among community dwelling elderly]. AB - OBJECTIVE: To characterize chronic pain and analgesic approaches in community- dwelling elderly of the northern district of Londrina - Brazil. METHODS: Cross sectional study with individuals 60 years old and more, resident in the local area of a Basic Health Unit (UBS) of Londrina, by home inquiry. Chronic pain was defined as pain lasting for six months or more. The elderly with pain, were questioned about characteristics of the pain as regards location, duration, frequency, intensity, improvement and worsening situations, as well as the impact of pain in the quality of life and also about analgesic approaches. RESULTS: One hundred and seventy two elderly were interviewed (101 women and 71 men). Presence of chronic pain was observed in 107 (62.%) of these 69.3% were female and 52.1% male (p = 0.004). The very old people, 80-year-old or more (p=0.01) and the depressive elderly (p=0.0008) presented higher frequency of chronic pain. Most prevalent pains were in the legs and back, with 31.2% each, and the majority of elderly referred to continuous, daily high intensity pain. Regarding analgesic approaches, the pharmacologic method was mentioned by 86 elderly (80.4%) and simple analgesic (32.6%) was the most used. CONCLUSION: Facts show that there is a high predominance of chronic pain in the population of elderly, mainly in women, in very old people and in depressive individuals. PMID- 18392485 TI - [Assessment of immunossupresion induced by thalidomide, cyclosporine and diclofenac on skin allograft survival in rabbits]. AB - OBJECTIVE: Allografting is one of the therapeutic alternatives for extensive burn victims without sufficient skin donor areas. This research studied the effects of cyclosporine, as an immunosuppressor model, and thalidomide and dyclofenac as anti-inflammatory drugs on an experimental skin allograft research. METHODS: Forty-two rabbits were divided in the following groups (n=6): Group 1 - autografting control; Group 2 - allografting control; Group 3 - allografts under thalidomide effect (100 mg/kg/day); Group 4 - allografts under sodium dyclofenac effect (2 mg/kg/day); - Group 5 -allografts under cyclosporine effect (10 mg/kg/day); Group 6 - allografts under cyclosporine effect (5 mg/kg/day); Group 7 allografts under cyclosporine (5 mg/kg/day) plus thalidomide (100 mg/kg/day) effect. Drugs were given via orogastric tube since the day before transplantation and daily during the postoperative period. Circular total skin grafts of the ear were exchanged between California and White New Zealand rabbits. RESULTS: Cyclosporine 10 mg/kg/day increased allograft survival and this effect was comparable to the association of cyclosporine 5 mg/kg/day with thalidomide 100 mg/kg/day. Thalidomide as an isolated drug and dyclofenac had a minimum effect on the average survival of the skin allografts. The number of eosinophils around the necrotic skin was higher in the dyclofenac group and lower in the group receiving cyclosporine associated with thalidomide. CONCLUSION: This study showed that thalidomide may be an useful drug when associated with subtherapeutic doses of cyclosporine for treatment of skin allografts. PMID- 18392486 TI - [Renal parenchymal disease: histopathologic-sonographic correlation]. AB - PURPOSE: This study was designed to address the correlation between sonography of a kidney with histological lesions and clinical findings in patients with renal parenchymal disease based on a multivariate logistic regression analysis. METHODS: Clinical and laboratory data, sonograms and renal biopsies were evaluated in 154 patients. Cortical echogenicity was graded as less than (0), equal to (1) or greater than (2) liver/spleen parenchyma. Histological lesions - mesangial proliferation (MP), leukocyte permeation (LP), fibrinoid necrosis and crescents (FNC), interstitial infiltrate (II), segmental glomerular sclerosis (SGS), glomerular obsolescence (GO), tubular atrophy (TA) interstitial fibrosis (IF) and interstitial edema (IE) - were graded according to extension and severity as normal (0%), mild (<25%), moderate (>25% <50%), and severe (>50%). RESULTS: a) II, IF, SGS, IE and increased creatinine occurred less in cortical echogenicity grade 0; b) MP, arterial hypertension and normal parenchymal thickness predict cortical echogenicity grade 1; c) IF, IE, increased creatinine and thin parenchyma predict occurrence of echogenicity grade 2; d) Excluding obese patients, both youth and hematocrit accounted for pyramid prominence; e) increased creatinine and GO was probable in patients with small kidneys. CONCLUSIONS: Increased cortical echogenicity was a very sensitive marker of renal parenchymal disease. Different lesions rather than degree of lesion severity accounted for progressive increase of cortical echogenicity. IE exponentially increased the effect of IF on cortical echogenicity. PMID- 18392487 TI - [Retiree quality of life assessment with SF-36 questionnaire]. AB - OBJECTIVE: Increase of the elderly in the Brazilian population demands assessment of aspects that interfere with the retirees' quality of life. METHODS: The Brazilian version of SF-36 questionnaire was applied to 87 retirees at their homes. Data on quality of life was associated with demographic data, socioeconomic status, health conditions and life style. Statistical analysis of data was performed using single-variable and multivariate analysis. RESULTS: The mean age was 57.3 years (standard deviation 8.9 years) and the mean retirement time was 7.1 years. Fifty-five percent were early retirements due to disability and 23.4% of those queried were still working when research was carried out. Smokers were 11.5% and 5.7 % were diagnosed as alcohol dependent by CAGE criteria. Depression and arterial systemic hypertension were the most prevalent conditions and 56.3% of the retirees practiced regular physical activity. Multivariate analysis disclosed that regular physical activity and post retirement occupation were the only variables associated with improved quality of life. CONCLUSION: The SF-36 questionnaire was a suitable instrument, relatively quick and easy to use. The quality of life was associated with life style, as indicated by the practice of physical activities and post-retirement occupation. Actions are needed to enable retirees to improve their life style after retirement. PMID- 18392489 TI - [HIV vaginal viral load in Brazilian HIV-infected women]. AB - OBJECTIVE: To evaluate factors associated to presence of free RNA-HIV in the vagina. METHODS: Cross-sectional study with HIV-infected women, excluding those who had undergone hysterectomy, had used vaginal medication within the last 48 hours, had had unprotected sex less than 72 hours before, were pregnant, or had genital bleeding. After signing an informed consent, blood samples were obtained for T CD4 lymphocytes count and plasmatic viral load, in addition to cervico vaginal lavage using 10 mL of sterile normal saline, later centrifuged, aliquoted and stored at - 70 degrees C to quantify free HIV-RNA. Plasmatic and vaginal viral load were measured using the kit HIV Monitor v1.5 Cobas Amplicor, Roche. Hybrid Capture test Digene was utilized for HPV (high and low risk), chlamydia trachomatis and N. gonorrhoea detection from an endocervical sample. Vaginal swab for bacterioscopy by the Gram method, evaluated according to Nugent criteria was obtained. RESULTS: Among 200 women evaluated, 73.5% were using HAART. The RNA-HIV was detectable in the vaginal lavage of 18 (9%), but in only one of those who had undetectable plasma viral load (0.5%). The vaginal prevalence of HIV was 24 times higher among those with detectable plasmatic HIV. Plasma viral load > 1500 copies/mL, no HAART use, reduced CD4 and bacterial vaginosis had increased prevalence of vaginal HIV-RNA, but in the adjusted statistical analysis, only the former remained significant CONCLUSION: Prevalence of vaginal HIV-RNA was low (9%). Plasmatic viral load > 1500 copies/mL, was the only risk factor for free vaginal HIV-RNA. PMID- 18392488 TI - [Polymorphisms GSTM1 and GSTT1 and sporadic breast cancer mammographic features]. AB - INTRODUCTION: Enzymes of the Glutathione S-transferase system (GST) modulate the effects of exposure to several cytotoxic and genotoxic agents. The GSTM1 and GSTT1 genes are polymorphic in humans and their deletions have been associated to increased risk of many cancers, including breast cancer. OBJECTIVE: To evaluate the occurrence of homozygous deletions of the GSTM1 and GSTT1 genes in women with sporadic breast cancer and in women without cancer and to compare breast cancer mammographic features between patients with and without these deletions. METHODS: The study evaluated 100 patients with sporadic breast cancer treated from September 2004 to June 2005 and 169 women without cancer, determining the frequency of the above-mentioned deletions by PCR and calculating the odds ratios and their 95% confidence intervals. Medical files and mammograms of 100 patients with breast cancer were evaluated and correlated with mammographic features such as density, mammographic findings and the BI-RADS classification. These findings were correlated with the genetic deletions by the PR (Prevalence-Ratio) with their respective 95% confidence intervals. RESULTS: The GSTM1 gene was deleted in 40% of the cancers and in 44.4% of controls (OR = 1.20; CI 95% 0.70 - 2.04; p=0.5659) while the GSTT1 gene was deleted in 20% and 19.5%, respectively (OR = 0.73; CI 95% 0.37-1.44; p=0.4124). High mammographic density had been associated with GSTM1 deletion (PR 2.43; CI 1.11 to 4.08). GST deletions were not associated with predominant mammographic findings and the BI-RADS classification. CONCLUSION: GSTM1 homozygous deletion was associated with high mammographic density. PMID- 18392490 TI - [Analysis of delays in diagnosis and treatment of breast cancer patients at a public hospital]. AB - BACKGROUND: To identify potential delays in the management of patients with breast cancer examined at a public hospital, from time of suspicion until diagnosis and beginning of treatment. METHODS: Retrospective study which analyzed data related to time elapsed in different intervals between suspicion, diagnosis and onset of systemic cancer-directed treatments, of consecutive breast cancer patients cared for at the medical oncology clinic of the Mario Covas Hospital during 2006. RESULTS: Sixty-eight women, with a mean age of 56.3 years (standard deviation: 12.2 years), were included. Of all the intervals, the longest delay occurred between the mammographic suspicion of cancer and performance of biopsy (median of 72 days, range: 4 - 1095 days); this was significantly longer (P<0.001) than in the other intervals analyzed. Furthermore, this interval in particular, was significantly longer in patients with advanced stage breast cancer when compared to those at the initial stage (P=0.014). CONCLUSION: Breast cancer patients treated in a public hospital in Brazil suffer delays, especially during the diagnosis of their disease. Minimizing the time between mammography and biopsy of suspicious lesions appears to be the most critical step to correct this situation. PMID- 18392491 TI - [The splenic inferior pole of rats and hyperbaric oxygen]. AB - OBJECTIVE: To study the functional and morphological features of the lower pole of the spleen in rats submitted, or not, to postoperative hyperbaric oxygen therapy. METHODS: Seventy-nine Wistar rats, weighing 248.7 +/- 27 g, divided into two groups [group A - simulation (n=40), group B - lower pole (n=39)] underwent surgery and were subdivided into two groups: 11 and 70 postoperative days. Each subgroup was subdivided into animals not treated (nt) (A11nt, n=10; B11nt, n=13; A70nt, n=10; B70nt, n=9) and treated with hyperbaric oxygen (t) (A11t, n=10; B11t, n=9; A70t, n=10; B70t, n=8). Blood was collected for measurement of lipids and immunoglobulins, platelet and Howell-Jolly body count before and after surgery. The spleen and lower pole were removed for histology. RESULTS: There was an increase of total cholesterol (p=0.002), VLDL-cholesterol and triglycerides (p=0.003) and of LDL-cholesterol (p=0.013) in subgroup B11nt, and no alterations were observed in the other subgroups. IgM decreased in subgroups B11t (p=0.007), B11nt (p=0.0000), B70nt (p=0.0005), B70t (p=0.02), and no change was observed in the simulation group. The number of platelets increased in subgroups B11nt (p=0.002) and B11t (p=0.01) and remained unchanged in the other subgroups. Howell Jolly bodies were less numerous in subgroup B70nt than in subgroup B11nt (p=0.019). Lower pole viability was better in subgroup B11t than in B11nt and in subgroup B70 than in B11, and did not differ between subgroups B70t and B70nt. CONCLUSION: Function and viability of the remaining lower pole improved during the late postoperative period. Viability and function of the lower pole were better during the early but not during the late postoperative period, in animals submitted to hyperbaric oxygen therapy. PMID- 18392492 TI - [From the molecular model to the impact on prognosis: an overview on acute promyelocytic leukemia]. AB - Acute promyelocytic leukemia (APL) is a model of clinical applicability of the knowledge of molecular physiopathology. It is characterized by recurrent genetic involvement of the retinoic acid alpha receptor. The consequence is a protein with low sensibility to its ligand and a myeloid maturation arrest. However, higher doses of all-trans-retinoic acid (ATRA) are able to supersede this deficiency and this is the mainstay of APL treatment leading to over 80% disease free survival, when adequately treated. Epidemiologically, it differs from other acute myeloid leukemia due to a higher incidence in young adults and in countries of "Latin" colonization. Differing from excellent results observed in developed countries, APL mortality in Brazil is still high, despite the wide availability of drugs. PMID- 18392493 TI - [Psychological assistance to medical students: 21 years of experience]. AB - The purpose of this article is to report on the 21-year experience of the Group of Psychological Assistance to Students at the School of Medicine of the Sao Paulo University (GRAPAL). First, the authors briefly introduce historical aspects of the psychological assistance provided to medical students, followed by the organization of GRAPAL in 1983 and the beginning of activities in 1986. Major reasons for medical students to seek psychological assistance, obstacles found by the institution, the work team and students are presented. Then, several works published by the Service, which approach the personality traits of medical students, their choice for Medicine and the suicide problem, are listed. Finally, the authors present some suggestions made by students to improve assistance services and measures to be taken by the GRAPAL team to carry out such suggestions. PMID- 18392494 TI - National Voice-Care Campaign and its importance in voice-care outreach activities. PMID- 18392495 TI - Complications of ent foreign bodies: a retrospective study. AB - Foreign bodies are one of the most common ENT (Ear, Nose and Throat) urgencies. Serious complications may occur, like tympanic perforations and bronchoaspiration, but they are uncommon. AIM: To analyze a 1356 foreign body series and establish causes for the complications, looking at prevention. MATERIALS AND METHODS: 1356 patients with ear, nose and throat foreign bodies from the ENT Department of Souza Aguiar Hospital, in Rio de Janeiro, between 1992 and 2000, were analyzed in a retrospective study for parameters like age, gender, type and localization of the foreign body, time span between introduction and removal of the foreign body and complications. RESULTS: The most common foreign bodies were beans and the most frequent age was between 1 and 4 years old. Ear foreign bodies were the most common, followed by nasal foreign bodies. Complications were statistically related to time, childs age and practical experience of the physician. CONCLUSION: Most of the situations related to ENT foreign bodies are avoidable. Improvements in Public Health Assistance and otolaryngologists training are essential to avoid serious complications. PMID- 18392496 TI - Incidence and evolution of nasal polyps in children and adolescents with cystic fibrosis. AB - Nasal polyps are a clinical sign of alert for investigating Cystic Fibrosis (CF). AIMS: To study the incidence of nasal polyps in children and adolescents with cystic fibrosis, its possible association with age, gender, clinical manifestations, genotype and sweat chlorine level, and its evolution with topical steroid therapy. METHODS: Clinical symptoms, sweat chlorine level and genotype were studied in 23 cystic fibrosis patients. Nasal polyps were diagnosed by nasal endoscopy and treated with topical steroids during 6 months, followed by a second nasal endoscopy. Fisher test was used for statistical analysis. RESULTS: Nasal polyps were found in 39.1% of the patients (five bilateral, four unilateral), all older than six years, recurrent pneumonia in 82.6%, pancreatic insufficiency in 87% and malnutrition in 74%. No association was seen between nasal polyps and sweat chlorine level, genotype, clinical sings of severity and nasal symptoms. Seven patients improved in their nasal polyps with topical steroids, six showed complete resolution. CONCLUSION: The study showed a high incidence of nasal polyps in older children, who span the entire range of clinical severity, even in the absence of clinical nasal symptoms. Topical steroid therapy showed good results. An interaction among pediatricians and otolaryngologists is necessary for diagnosis and follow-up. PMID- 18392498 TI - Flexible nasolaryngoscopy accuracy in laryngomalacia diagnosis. AB - Laryngomalacia is the most common cause of stridor in infancy. Diagnosis is established by associating the clinical manifestations with laryngoscopic findings in a dynamic form. AIM: To analyze diagnostic accuracy of laryngomalacia through flexible nasolaryngoscopy performed by four different examiners. Form of studying: Comparison of diagnostic tests (clinical study). MATERIAL AND METHOD: A protocol of videolaryngoscopic evaluation for patients with laryngomalacia was created encompassing the following items: anterior displacement of the arytenoids; omega-shaped epiglottis; short aryepiglottic folds; posterior displacement of epiglottis; vocal folds being visible or not; edema of the posterior larynx. Four different examiners analyzed the videolaryngoscopic examinations of 18 patients with established diagnosis of laryngomalacia previously established by clinical data, epidemiology and anatomical traits. The four observers knew previously that the patients had laryngomalacia and which criteria should be analyzed in the evaluation protocol. Observers were unaware of the results each one found. RESULTS: diagnostic agreement average considering all parameters evaluated was of 88.2%. DISCUSSION/CONCLUSION: Dynamic flexible nasolaryngoscopy is a proven diagnostic method, regardless of physician experience. PMID- 18392497 TI - Hearing loss in congenital toxoplasmosis detected by newborn screening. AB - Congenital toxoplasmosis may cause sensorineural deficit in up to 20% of the patients and proper treatment in the first year improves prognosis. In Brazil, this infections impact on hearing impairment is unknown. AIM: To evaluate hearing of newborns with congenital toxoplasmosis identified by the newborn screening service. METHOD: This prospective study analyzed children with congenital toxoplasmosis identified by newborn screening (IgM anti-T.gondii) in Belo Horizonte during 2003/2004. The presence of IgM and/or IgA in the first 6 months or IgG at 12 months-of age in serology was used as case definition. Hearing tests were carried out at the time of diagnosis and 12 months later, including behavioral audiometry, evoked otoacoustic emission and brainstem evoked responses audiometry. RESULTS: Among 30.808 screened children (97% of live births), 20 had congenital toxoplasmosis, 15 (75%) were asymptomatic at birth. Nineteen children were evaluated by hearing tests. Four had sensorineural impairment (21.1%). One child had other risk factors for hearing impairment; the other three had no other risk factors but toxoplasmosis. Two properly children treated still had hearing loss, in disagreement with current literature. CONCLUSION: Results suggest that congenital toxoplasmosis, common in Brazil, is a risk factor for hearing impairment and its impact on hearing loss deserves further studies. PMID- 18392499 TI - Temporo mandibular joint ankylosis. AB - Ankylosis may be defined as joint surfaces fusion. The treatment of temporomandibular joint ankylosis poses a significant challenge because of the high recurrence rate. AIM: The aim of this study is to report six cases treated by joint reconstruction, evaluate the results of these surgeries and review the literature. METHODS: The sample in this retrospective study was obtained from the records of the university hospital, patients who had to undergo ankylosis treatment by alloplastic or autogenous graft between March 2001 and October 2005. Pre - and post-operative assessment included a throughout history and physical examination to determine the cause of ankylosis, the Maximum mouth opening (MMO), etiology and type of ankylosis, recurrence rate and presence of facial nerve paralysis. RESULTS: The mean MMO in the pre-operative period was 9.6 mm (0 mm to 17 mm) and in the post-operative period it was of 31.33 mm (14 mm to 41 mm), there was no facial nerve paralysis and there was recurrence in just one case. CONCLUSION: The joint reconstruction with alloplastic or autogenous grafts for the ankylosis treatment proved to be efficient in relation to the post-operative MMO, recurrence and joint function. PMID- 18392500 TI - Nasolabial cyst: diagnostic and therapeutical aspects. AB - Nasolabial cyst is a rare lesion situated behind the ala nasi, extending backwards into the inferior nasal meatus and forward into the labio-gingival sulcus. AIM: We present our case of a nasolabial cyst, with the purpose of discussing clinical presentation, diagnosis and the more suitable surgical techniques to treat this disorder. MATERIALS AND METHODS: A retrospective study of eight patients with diagnosis of nasolabial cyst, carried out in the period of january/2000 to december/2006. The diagnosis was suggested by otorhinolaryngology exam and computer tomography. All patients were submitted to surgical treatment (enucleation) and definitive diagnosis was confirmed by histopathology. RESULTS: Predominant symptoms were nasal obstruction, swelling in the nasal vestibule region and local pain. Patients had had symptoms for a median of 26.2 months. CT scan was performed in all patients, showing a well outlined cystic lesion with bone remodeling in some cases. Median sizes of the cysts were 2.18 cm. There was no evidence of recurrence during a mean follow-up of 19.5 months. CONCLUSION: Nasolabial cysts are rare lesions. Common presentation is a well-confined swelling, local pain and nasal obstruction. Enucleation is the treatment of choice with low recurrence rate. PMID- 18392501 TI - Quantitative analysis of myelinic fibers in human laryngeal nerves according to age. AB - INTRODUCTION AND AIM: To carry out a morphometric analysis of myelinic fibers in laryngeal nerves aiming to identify quantitative changes as a result of aging. STUDY DESIGN: Clinical and experimental. MATERIAL AND METHOD: A 1cm fragment was collected from the superior laryngeal nerves and recurrent laryngeal nerves taken from twelve male cadavers. The sample was divided into two groups: those aged below 60 years (Adult) and those aged 60 years or more (Elderly). The material was evaluated under light microscopy coupled with an image analysis system. RESULTS: The total number of myelinic fibers from the superior laryngeal nerve was similar in both age groups; there was, however, a trend for a higher number of 1 microm fibers in the adult group (p=0.0744). The adult group had a higher total number of myelinic fibers in the recurrent laryngeal nerve (p=0.0006), and this difference was seen in fibers with diameters between 1-3 microm (p<0.007). The adult group had a higher total number of myelinic fibers in the laryngeal nerves (sum of superior laryngeal nerves and recurrent laryngeal nerves fibers) compared to the elderly group (p<0.0091). CONCLUSION: The total number of myelinic fibers in laryngeal nerves is higher for the group aged below 60 years. PMID- 18392502 TI - Effect of trans-retinoic acid in the inhibition of cholesteatoma in guinea pigs. AB - Middle ear cholesteatoma affected more than 5 million people until the 80;s. Many animal models were used, unsuccessfully, to study an alternative therapy to cholesteatoma. AIM: observe the effect of the trans-retinoic acid in the inhibition of middle ear cholesteatomas induced by propylene glycol. STUDY DESIGN: Clinical and Experimental. METHODS: 25 guinea pigs were submitted to the application of a 100% propylene glycol solution in their bulla bilaterally and a solution of trans-retinoic acid was applied locally in the external right ear, while in the left ear saline solution was applied (control ear). The guinea pigs were slaughtered and their temporal bones were prepared for macroscopic and histological analysis. RESULTS: The macroscopic findings had evidenced the presence of cholesteatoma in 25% of the right ears and 85% of the left ears (P=0.0003 *). The histological study had evidenced the presence of cholesteatoma in 30% of right ears and 75% of the left ears (P=0.0104*). CONCLUSION: The local use of the trans-retinoic acid is effective in inhibiting the induced formation of cholesteatomas in guinea pigs. PMID- 18392503 TI - Evaluation of a new mouthwash on caseous formation. AB - Tonsil caseous affects a significant percentage of the population. Surgeries, conservative or not, have been the only viable alternatives of treatment. However, today there is still not, up to now, an economical and non-invasive treatment that presents satisfactory results. AIMS: The objectives of this study are to evaluate the efficiency of a mouthwash, with active ingredients that associate oxygenating and antimicrobial substances, in the reduction of caseous and tongue coating formation, whose etiology is similar to caseous, and to evaluate the reduction of the volatile sulfur compounds (VSCs) concentration. STUDY DESIGN: Double blind, placebo-controlled, randomized, clinical and experimental study. MATERIAL AND METHODS: [corrected] A sample of 50 volunteers with more than one year of chronic caseous tonsillitis complaint used it. The research was carried out in 2005, in the cities of Sao Paulo and Campinas. RESULTS: For the group that used the placebo solution, there was no correlation between the variables or statistical significance in the results. For the group that used the mouthwash, the results were significant in all analyzed questions. CONCLUSIONS: This new mouthwash proved to be a viable conservative alternative for the treatment of tonsil caseous, being also efficient in the reduction of tongue coating formation and VSCs concentration. PMID- 18392504 TI - Epidemiologic evaluation of head and neck patients in a university hospital of Northwestern Sao Paulo State. AB - Head and neck cancer accounts for nearly 200.000 new cases worldwide. A mean of 13.470 new cases of cancer in the oral cavity for 100.000 inhabitants is observed in Brazil. AIM: To analyze clinical and epidemiological aspects in patients consulted in the Otorhinolaryngology and Head and Neck Surgery ward in a University hospital of Northwestern Sao Paulo, Brazil. MATERIALS AND METHODS: A total of 427 patients consulted in the hospital in the period from 2000 to 2005 were investigated. The variables analyzed included: age, gender, occupation, skin color, tobacco and alcohol consumption, primary site of the tumor, clinical staging, degree of histological differentiation and outcome. The data was analyzed by descriptive and exploratory statistics. RESULTS: Prevalence was found among men (86%), white color (90%), smokers (83.37%), and alcoholics (65.80%); the average age was 61 years, 24.25% of men were farmers and 60% of women, housekeepers. Primary site of tumor was usually in the oral cavity (35.37%), with histological squamous cell. The incidence of deaths was 164. CONCLUSION: This study has provided the profile of the patients assisted in this hospital; moreover, it has contributed to outline further programs for preventing this disease. PMID- 18392505 TI - The importance of pre-epiglottis space invasion in the treatment planning of larynx and hypopharynx cancer. AB - The involvement of pre-epiglottis space can change the indication for partial laryngeal resection. AIM: The aim of this study was to evaluate inter-observer and intra-observer agreement by means of computed tomography analysis regarding the involvement of the pre-epiglottis space (PES) from carcinoma of the upper aerodigestive tract and its relation with therapeutic planning. MATERIALS AND METHODS: Retrospective study of ninety-five computed tomography exams of patients with squamous cell carcinoma, from 1990 to 2004, were selected and evaluated; 87 were males and eight females, with ages ranging from 32 to 73 years. Imaging results were analyzed twice by three radiologists, individually, without any previous knowledge of the clinical stage. No patient had received any previous treatment up to the moment of imaging examination, such as surgery, chemotherapy or radiotherapy. All the cases were confirmed by biopsy. Information was obtained from the medical charts. RESULTS: Kappa Index was calculated by assessing agreement between the three observers. We obtained substantial to almost perfect levels of agreement. CONCLUSIONS: After a general Kappa Index of 0.72, the results suggest a substantial agreement in the involvement of the PES by means of computed tomography analysis. PMID- 18392507 TI - Oroantral fistula and genian mucosal flap: a review of 25 cases. AB - The oroantral fistula is a pathological connection between the maxillary sinus and with the oral cavity. The condition mostly follows dental extraction. AIM: To present the experience of 25 cases. MATERIAL AND METHODS: Retrospective cases between 1996-2000. The ORL examination included nasal or sinusal endoscopy, a CT scan and histopathological analysis. RESULTS: Twenty-five cases were found: ten 2nd molar cases, eight 1st molar cases, six 2nd premolar cases, and one canine case. All patients underwent a Caldwell-Luc operation plus excision of the epithelium lining the fistula, that was then completely covered by a flap of mucosa rotated from the genian region. DISCUSSION: In cases of major fistulae a bone autograft taken from the anterior sinus wall was used. Bacterial cultures (n=19) revealed streptococus pneumoniae (13), haemophillus influenza (6), Moraxella catharralis (2) and staphylococus aureus (2). Aspergillus niger was found in one case presenting as a "fungic ball". CONCLUSIONS: The only case of surgical failure, after 30 days postoperatively, was reoperated, using a bone graft. After a 6-month follow up all of the patients progressed satisfactorily, including the reoperated patient. PMID- 18392506 TI - Assessment of carotid artery invasion by lymph node metastasis from squamous cell carcinoma of aero-digestive tract. AB - Carotid involvement by metastatic lymph nodes changes the approach in head and neck tumors. AIM: To evaluate interobserver agreement by CT scan analysis regarding the involvement of the carotid artery by metastatic lymph nodes from squamous cell carcinoma of the upper aerodigestive tract and its relation to resection criteria. MATERIALS AND METHODS: retrospective study of 99 CT images of patients with SCC were evaluated. Eighty-six were males and 13 females, with ages ranging from 32 to 76 years. Four radiologists, without any previous knowledge of the clinical stage, analyzed imaging results. No patients had received previous treatment and histological diagnoses were obtained through biopsy. The carotid artery invasion was classified as simple (0 to 50% and from to 100%), and complex (0 to 25%; 26 to 50%; 51 to 75% and 76 to 100%). The level of interobserver agreement was obtained through Kappa Index (p < 0,05) and the concordance power varied from despicable to excellent. RESULTS: The Kappa Index were moderate (0, 53%) for simple classification and minimum (0, 36%) for complex classification. CONCLUSIONS: The computed tomography showed low effectiveness in the evaluation of lymph node metastasis resection concerning carotid artery invasion. PMID- 18392508 TI - Comparison between the IT-MAIS and MUSS questionnaires with video-recording for evaluation of children who may receive a cochlear implantation. AB - There is a great difficulty in determining earlier on which children would benefit or not from cochlear implants, especially because of their young age, the responses they give are very subtle. AIM: To compare results obtained through video-recording of the interactions of children who may receive a cochlear implant with the results obtained through evaluation protocols. METHOD: Seven children, with an average age of 39.7 months, with profound hearing loss were selected for the study. IT-MAIS and MUSS questionnaires were given to their parents/guardians of these children and the results were compared with the observation of the video-recordings. RESULTS: It was possible to observe that the data is compatible with the auditory stages. However, the MUSS questionnaire data gathered during playful activities is very different . The questionnaire only takes into consideration the use of verbal language and therefore the majority of the evaluated children inevitably score low. CONCLUSION: Observing children play allows us to trace a better profile of linguistic behavior and aspects relative to language, that may presented differences in the questionnaire. PMID- 18392509 TI - Comparison between general anesthesia and superficial cervical plexus block in partial thyroidectomies. AB - Thyroidectomy under the effect of superficial cervical plexus block (SCPB) has met resistance. AIM: to compare variables in patients submitted to hemithyroidectomy under the effect of general anesthesia (GA) and SCPB. CASE REPORT AND METHODS: GA was used in 21 patients, and SCPB was used in another 21 patients. Following sedation, marcaine 0.5% with vasoconstrictor was used in the SCPB group. Intraoperative sedation with diazepam and metoprolol to control arterial pressure and cardiac frequency was given as needed. GA followed the standard method in the unit. RESULTS: We found significant results (p<0.05, Students t-test) for surgery time (GA - 111.4 min; SCPB - 125.5 min), anesthesia time (GA - 154.1 min; SCPB - 488.6 min), time in the surgery room (GA - 15 min; SCPB - 1 min), treatment costs (GA - R$203.2; SCPB - R$87.4), presence of bradycardia (GA - 0; SCPB - 23.8%) and laryngotracheal injury (GA - 51; SCPB - 0 %). We also found the following non-significant results: hospitalization time (GA - 17.3; SCPB - 15.1 hours); bleeding volume (GA - 41,9 g; SCPB - 47.6 g), size of the operative specimen (GA - 52.1 cm3; SCPB - 93.69 cm3) and patient satisfaction level (GA - 3.8; SCPB - 3.9). CONCLUSION: Although the incidence of bradycardia was higher (23.8%), SCPB was done for the resection of tumors measuring up to 348 cm3, at a lower cost and with no laryngotracheal injuries; these were present in 51% of patients undergoing GA. PMID- 18392510 TI - Study of otoacoustic emissions during the female hormonal cycle. AB - The hormonal changes that occur in a short time span promote modifications all over the woman's body, with physical and emotional manifestations which are frequently observed. AIM: to evaluate the activity of the external ciliated cells in women during their menstrual cycle, observing the effect of hormonal changes caused by the cycle in their 3 phases. METHODS: this is a longitudinal prospective study where 21 women between 20 and 35 years old who did not take any contraceptive medicine were assessed. Transient otoacoustic emissions were evaluated by distortion product during the 3 phases of the menstrual cycle (luteal, follicular and ovulatory phases). The SPSS 13.0 software was used to analyze the data. RESULTS: the phases of menstrual cycle do not alter the amplitude and reproducibility values of the transient otoacoustic emissions. We noticed a difference between the ears in the frequency of 1.5 KHz in the amplitude of emissions by distortion product, and the right ear showed the highest values. CONCLUSION: There are no significant differences in transient otoacoustic emissions and distortion products in the phases of the menstrual cycle. PMID- 18392511 TI - Comparative study between school performance on first grade children and suppression of otoacoustic transient emission. AB - School learning can be hampered if there are defects on the central auditory process. Since those with auditory deficiency can be rehabilitated, it is fundamental that we identify them. Otoacoustic emissions test has low cost and operational ease. STUDY DESIGN: clinical and experimental. AIM: to study the relationship between school learning and transient otoacoustic emission suppression by contralateral stimuli. MATERIAL AND METHODS: 39 individuals, from 7 to 12 years of age were evaluated, 19 (48.7%) with good school performance and 20 (51.3%) poor performers. RESULTS: A transient otoacoustic emission suppression failure for contralateral acoustic stimuli was more frequently found among children with poor school performance. We established a value of 1.6 dB SPL for emission reduction that characterized those children as belonging to the poor learning performance group: sensitivity 65%, specificity 72,2%, accuracy of 68.4%, positive predictive value of 72.2%. CONCLUSION: The contralateral emission suppression test of the right ear can be predictive of school difficulties in individuals from six to twelve years of age. PMID- 18392514 TI - Extratympanic electrocochleography in the diagnosis of auditory neuropathy/auditory dyssynchrony. AB - The brainstem auditory evoked potential (BAEP) is being extensively used as a method for the evaluation of cochlear function in individuals with diagnosis of auditory neuropathy/auditory dyssynchrony (AN/AD). In the absence of otoacoustic emissions, many cases of AN/AD have been diagnosed by the presence of CM identified in the BAEP. AIM: to demonstrate the clinical applicability of extratympanic electrocochleography (ET-Ecochg) in the differential diagnosis of AN/AD compared to the BAEP. METHOD: a 4-year-old child with a diagnosis of AN/AD seen at the Audiological Research Center was submitted to ET-Ecochg with a 2000 Hz tone burst in rarefaction and condensation polarities. RESULTS: the ET-Ecochg exam was illustrated. Using an appropriate protocol, it was possible to demonstrate CM and to confirm it in the Ecochg, with a recording quality superior to that obtained in the BAEP. CONCLUSION: ET-Ecochg permitted a more detailed analysis of CM compared to the BAEP, thus showing clinical applicability for the investigation of cochlear function in AN/AD. PMID- 18392513 TI - Mobile phones: influence on auditory and vestibular systems. AB - Telecommunications systems emit radiofrequency, which is an invisible electromagnetic radiation. Mobile phones operate with microwaves (450900 MHz in the analog service, and 1,82,2 GHz in the digital service) very close to the users ear. The skin, inner ear, cochlear nerve and the temporal lobe surface absorb the radiofrequency energy. AIM: literature review on the influence of cellular phones on hearing and balance. STUDY DESIGN: systematic review. METHODS: We reviewed papers on the influence of mobile phones on auditory and vestibular systems from Lilacs and Medline databases, published from 2000 to 2005, and also materials available in the Internet. RESULTS: Studies concerning mobile phone radiation and risk of developing an acoustic neuroma have controversial results. Some authors did not see evidences of a higher risk of tumor development in mobile phone users, while others report that usage of analog cellular phones for ten or more years increase the risk of developing the tumor. Acute exposure to mobile phone microwaves do not influence the cochlear outer hair cells function in vivo and in vitro, the cochlear nerve electrical properties nor the vestibular system physiology in humans. Analog hearing aids are more susceptible to the electromagnetic interference caused by digital mobile phones. CONCLUSION: there is no evidence of cochleo-vestibular lesion caused by cellular phones. PMID- 18392512 TI - In vitro antimicrobial activity of plants in Acute Otitis Externa. AB - Acute Otitis Externa is an inflammation of the outer auditory meatus, and according to popular saying, medicinal plant extracts can be used in its treatment. AIM: to assess the in vitro antimicrobial activity of the following plants: Aleolanthus suaveolens; Caryophyllus aromaticus; Cymbopogon citratus; Matricaria chamomila; Pithecellobium avaremotemo; Plectranthus amboinicus and Ruta graveolens on the germs that cause otitis externa. MATERIALS AND METHODS: the minimum inhibitory concentration of extracts and oils from these plants was obtained from otitis externa samples. RESULTS: Staphylococcus aureus in 10 cultures, Pseudomonas aeruginosa in 8, Pseudomonas aeruginosa and Staphylococcus aureus together in 5 cultures and Candida albicans and Candida krusei in 4 cultures. P. aeruginosa was resistant to all oils and extracts tested; extracts from A. suaveolens, P. avaremotemo and R. graveolens were inactive; the essential oil from C. aromaticus and M. chamomila were active against 3 strains of S. aureus and the Candida strains; seven of the S. aureus strains were sensitive to the P. amboinicus extract; however, the oil was inactive against 4 S. aureus strains and the Candida strains were sensitive to the R. graveolens essential oil. CONCLUSION: depending on the etiological agent, some plants presented satisfactory results, however we still need more detailed studies in order to better use these plants. PMID- 18392515 TI - Foreign body in the Eustachian tube: case presentation and technique used for removal. AB - Foreign bodies of the external ear are very common; the same can not be said about foreign bodies of the middle ear, especially of the Eustachian tube. CASE PRESENTATION: Alcoholic and psychopathic patient presented a foreign body (barbecue wooden stick) purposefully introduced in his middle ear and Eustachian tube during an act of delinquency. The foreign body was stuck in the tube and could not be removed externally. It was surgically removed as described by the authors later on in the paper. COMMENTS: The patient recovered well, with no sequela on the facial nerve and without important vertigo. PMID- 18392516 TI - Laryngocele: a cause of upper airway obstruction. AB - Laryngoceles are abnormal dilatations of the laryngeal saccule, which rises between the ventricular folds, the base of the epiglottis and the inner surface of the thyroid cartilage. Clinical symptoms are rare, and the find of asymptomatic laryngoceles in pathology studies are frequent. Sometimes it is presented as cervical swelling causing airway obstruction in need of emergency intervention. In this study, we report a case of upper airway obstruction due to laryngocele treated by emergency tracheotomy and we review of the literature. Laryngocele complications include infection (pyocele formation), pathogens aspirations with subsequent bronchitis and pneumonia and upper airway obstruction, as in the case reported. Despite being benign tumors, laryngoceles cause relevant airway obstruction. Correct diagnosis and proper treatment can preclude emergencies as it happened to our patient hereby reported. PMID- 18392517 TI - Nasal cavity vascular leiomyoma: case report and literature review. AB - Liomyomas of the nasal cavity and paranasal sinuses are rare. They make up less than 1% of all leiomyomas in the human body. This is due to the paucity of smooth muscle in the nose. They are classified in three groups: leiomyoma, angiomyoma and epithelioid leiomyoma. Only 15 cases of vascular leiomyomas have been found in the literature. The treatment of choice is surgical excision. Hereby we present a new case and review the literature. PMID- 18392519 TI - Mandibular ameloblastoma treated by bone resection and immediate reconstruction. AB - Multicystic ameloblastoma mainly affects adult patients between the third and seventh decades of life, frequently in the posterior region of the mandible. The resection of a mandible segment without adequate reconstruction produces serious esthetic and functional sequelae leading to a loss of quality of life. The objective of this study is to show that multidisciplinary treatment of ameloblastomas helps in total lesion excision associated with complete reconstruction of the damaged area. We present a 47-year-old male patient with an ameloblastoma in the posterior mandible who was treated with complete resection of a mandibular segment. Reconstruction, carried out during the same surgical procedure, was performed using an iliac crest bone graft fixed with titanium plates and screws. Rehabilitation was completed eight months later with teeth implants in the grafted area. The advantages of this procedure include recurrence risk reduction due to segmental resection, reliable mandibular reconstruction and less surgical procedures, allowing full rehabilitation within a shorter period of time. PMID- 18392518 TI - Inverted papilloma in the sphenoethmoidal recess. AB - Inverted papilloma is a nasal benign tumor that usually arises from the lateral nasal wall, especially from the middle meatus. It has high local invasive likelihood, high recurrence rates and malignancy potential. Sphenoethmoidal recess involvement is rare and is usually due to sphenoid sinus primary disease. In the literature, no case of isolated sphenoethmoidal recess inverted papilloma has been reported yet. The present report describes an exceptional location of inverted papilloma, arising from the sphenoethmoid recess, without involving the sphenoid sinus. PMID- 18392520 TI - Intracanalicular meningioma: diagnostic by immunohistochemistry. PMID- 18392521 TI - Temporal bone histopathology: idiopathic sudden hearing loss. PMID- 18392522 TI - Angiolymphoid hyperplasia with eosinophilia: excellent response to intralesional triamcinolone. PMID- 18392523 TI - [The Latin American Journal of Nursing celebrates its 15th anniversary]. PMID- 18392524 TI - Quality of life from the perspective of revascularized patients during rehabilitation: an ethnographic study. AB - This ethnographic study aimed to understand the meaning of quality of life from the perspective of people with coronary disease submitted to myocardial revascularization surgery. Direct observations and semi-structured interviews were used for data collection. Data were analyzed through careful reading, identification of units of meaning, thematic groups and themes, and interpreted according to interpretative anthropology and the Centre for Health Promotion conceptual model of quality of life. The identified theme, quality of life being happy as possible, is based on being healthy, having a job and a harmonious family. These were considered dimensions for a "tranquil and happy" life. The meaning of quality of life is complex, subject to new significations throughout life, and related to the socioeconomic and cultural context, including, in the case, the experience of the health-disease process. PMID- 18392525 TI - Between freedom and reclusion: social support as a quality-of-life component in the family caregiver-dependent person binomial. AB - This is a qualitative study which uses Grounded Theory as its methodological framework and Symbolic Interactionism as a theoretical base to understand the experience of family caregivers for Cerebrovascular Accident (CVA) patients with regard to social support during their rehabilitation process at home. The components (themes and categories) of the phenomenon assuming home care and specifically the themes assuming care with support and assuming care without support were inter-related for the purpose of comparison and analysis, in order to apprehend how the interaction between them occurred. It was observed that, in addition to the recovery of the patient's autonomy, social support is one of the intervenient components in the quality of life for the family caregiver-disabled person binomial, particularly with respect to the caregiver's freedom to resume his/her life plan. PMID- 18392526 TI - Stressing factors and coping strategies used by oncology nurses. AB - In the oncology specialty, many factors can result in occupational stress in nursing professionals. As an attempt to controlling this situation, individuals may use coping strategies. Coping is a cognitive and behavioral effort one uses to face a stressful situation. The aims of this study were to identify the stressful factors regarding oncology nurses, and to verify what coping strategies they use. Two questionnaires were used: a demographic data inventory, designed by the researcher, and the Folkman and Lazarus coping strategies inventory. The results showed that the main stressful factors for oncology nurses are patient death (28.6%), emergency situations (16.9%), relationship issues with the nursing team (15.5%), and work-process situations (15.5%). In the studied population, the main coping strategy used was positive reappraisal. PMID- 18392527 TI - Stress in nurses working in intensive care units. AB - The present study aimed to identify the presence of stress in nurses working in intensive care units, the stressing agents and symptoms associated to the nurses' perceptions of stress, and to assess the correlation between the occurrence of stress, sources of stress, and symptoms shown by the nurses. Seventy-five nurses took part in the study. The data were collected from questionnaires, analyzed with the Pearson correlation coefficients, and adjusted by general linear models. The study showed the presence of stress related to work dissatisfaction, activities regarded as critical situations in intensive care units, symptoms related to cardiovascular, digestive and musculoskeletal disorders. The conclusion is that stress is present in nurses' activities in intensive care units, related to characteristics of the health area itself, causing dissatisfaction and stress-related symptoms. PMID- 18392528 TI - Effects of relaxation on anxiety and salivary IgA levels in puerperae. AB - This experimental study aimed to evaluate the effect of relaxation techniques on anxiety levels, and the relation between anxiety and the concentration of Immunoglobulin A. The study was carried out in a maternity hospital in a city of the State of Espirito Santo, Brazil. The sample was composed of 60 puerperae. The information on the variables: age, education, marital status, type of childbirth, and parity were collected with a specific form; the trait and state of anxiety were based on the State Trait Anxiety Inventory (STAI/IDATE); and the level of salivary IgA was obtained through immunoturbidimetry. The application of the Mann Whitney, Wilcoxon, and Pearson's correlation statistical tests showed a significant reduction in the levels of the state of anxiety in the experimental group (p = 0.01); there was no correlation between the trait and state variables of anxiety and the salivary IgA level; both groups (experimental and control) showed trait and state of medium-intensity anxiety. PMID- 18392529 TI - Cost-effectiveness analysis of the analgesic therapy of postoperative pain. AB - The study aimed to compare cost-effectiveness of analgesic schemes administered to 89 patients submitted to hemorrhoidectomy, on the 1st postoperative day. The descriptive and retrospective study was carried out in a General Hospital, Sao Paulo, Brazil. In order to carry out the cost-effectiveness analysis, the five most frequently used analgesic schemes were identified in practice. The main outcome was the absence of breakthrough pain episodes. While calculating the costs, analgesics and all devices related to the schemes were taken into consideration. Codeine 120 mg+acetaminophen 2000 mg was the most effective therapy with the lowest cost per patient with no breakthrough pain episodes ($65.23). Incremental analysis indicated that codeine 120 mg+acetaminophen 2000 mg+ketoprofen 200 mg involved the additional cost of $238.31 in case an extra effectiveness benefit was needed. The analysis showed that the most suitable choice of analgesic therapy should consider the resources available at the institution along with economic and clinical aspects. PMID- 18392530 TI - Development of an eye protector for phototherapy on newborns: a technology. AB - Hyperbilirubinemia results from the predisposition of newborns to produce bilirubin and their ability to excrete it. The treatment applied is phototherapy; however, one of its complications is degeneration of the retina by exposure to light, making it essential to provide proper eye protection. The objective of this study is to describe the development of an invention (utility model patented) for eye protection in newborn phototherapy. This invention aims to replace the current widespread practice of using improvised means that are uncomfortable for the baby and disrupt the mother/child relationship. A study was made of the radiances emitted by light sources used in phototherapy. The invention consists of an eye protector model for use during the newborn's phototherapy, allowing greater closeness between mother and baby. The device is easy to use, economical, removable and appropriate for newborns, causing no discomfort, and its effectiveness has been proved through radiance tests. PMID- 18392531 TI - Delay factors on the administration of thrombolytic therapy in patients diagnosed with acute myocardial infarction in a general hospital. AB - OBJECTIVE: To identify factors that delay the onset of thrombolysis in patients with acute myocardial infarction (AMI). METHODS: A cohort study was carried out with 146 patients, each diagnosed with AMI and subjected to thrombolytic therapy. The data was extracted from medical records between January 2002 and December 2004. RESULTS: The average age of the studied population was 57.5 +/- 9 years, 64.4% were male. The average time between the onset of pain and arrival at the hospital was 254.7 +/- 126.6 minutes, 28.1% used an ambulance for the trip to the hospital, the door-to-electrocardiogram time averaged 19.4 +/- 7.3 minutes and the door-to-needle time was 51.1 +/- 14.9 minutes. There was no significant difference between the time of arrival to the hospital and the method of transportation used (P= 0.81), and those seen by cardiologists and during the nightshift had a reduction in the door-to-needle time, respectively (P=0.014) and (P=0.034). CONCLUSIONS: Study results show that the delay in the search for medical service, and the long time taken from door-to-electrocardiogram and to reach the AMI diagnosis were the factors involved in the delay of thrombolytic treatment. PMID- 18392532 TI - Quality assessment of the wound dressing procedure in patients at a university hospital. AB - This observational and sectional study analyzed the quality of the wound dressing procedure performed on hospitalized patients at a medical surgical unit of a University Hospital, based on their classification according to the degree of care dependency and activity performance phases. Using a check list, 168 wound dressings were observed between October and December 2005. Procedure quality was analyzed based on the Positivity Index (IP) and values >70% were considered satisfactory. For the preparation, the IP was 68%, 63%, 73% and 75% for patients with degrees I, II, III and IV, respectively; for execution, 70%, 69%, 71% and 75% and, for unit organization, it was >70% for all degrees. However, the items: validity time frame checking, respect for aseptic principles and maintenance of logical sequence of procedures were compromised. Rigorous execution of procedures allows for risk decrease and assures beneficial results for patients, conferring quality to nursing actions. PMID- 18392533 TI - Verbal contents expressed by mothers of preterm infants with clinical emotional symptoms. AB - The aim of the present study was to characterize verbal contents expressed by preterm neonates' mothers during psychological support intervention. The sample was composed by 20 mothers of pre-term and very low birthweight neonates, hospitalized in NICU, allocated into two groups: 10 mothers with emotional clinical symptoms (MECS) and 10 mothers with no symptoms (MNECS), assessed by the Beck Depression Inventory and the State-Trait Anxiety Inventory. The verbal behavior expressed by mothers in the psychological support group was recorded, transcribed and categorized. The results show that three verbal categories were more frequent: negative feelings or reactions, followed by positive feelings or reactions and communication with the health staff. The comparison between groups reveals that MECS presented more expressions in the first category in comparison to MNECS. The identification of the feelings and reactions of pre-term neonates' mothers and emotional clinical symptoms should be included for appropriate intervention in developmental care in NICU. PMID- 18392534 TI - Background and managerial practice of nurses: paths for transforming praxis. AB - Qualitative research using a dialectics view; the objective is to analyze the demands and the expectations from the setting of managerial practices in view of the Pedagogical Project and the strategies which may or may not favor the transforming praxis. It involves 11 students, 12 professors, and 12 service nurses who are the participants in the teaching-learning process of managerial knowledge, in the interceding space, in the fields of practice and in the curricular training programs, both in the hospital environment and in the basic health services and of the Nursing graduation course from a Federal University, from the city of Alfenas-MG. It uses the collecting technique by focal group and the analysis of the contents is subsided by two great discussions: background and work. The results point that the nurse's background and praxis must go through the work processes in the caring, managerial and educative dimensions as well as in the scientific investigation, building paths for the development of competences, in the teaching and service interaction, following the current National Curriculum Guidelines. PMID- 18392535 TI - Creation and evaluation cycle of a distance module for nursing undergraduates, named "medication administration". AB - The objective of this study was to describe the creation cycle of the module "medication administration" and to evaluate it. It is an evaluation research, using the Engagement Theory as a methodological/theoretical approach carried out in two phases: the first related to the creation of the module and the second to its evaluation by specialists using the instrument based on the Model of Analytical Appreciation of Hypermedia Systems. The creation cycle of the module followed the following steps: gathering the objectives of the module, organization of the content, selection of the media, creation of the classes and activation of the learning process. It was evaluated by contents specialists, informatics specialists and nursing students. The evaluation of the distribution and the accessibility of the contents, the utilization of images, the clarity and ease of execution of the program were positive. The module was considered appropriate to be used with nursing students. PMID- 18392536 TI - Superficial peripheral vein type classification of adolescents, adults and elderly according to the Delphi technique. AB - This descriptive research attempts to cooperate with the standardization of communication about vein types in nursing. Authors utilized the "Delphi" technique and aimed at elaborating and validating a peripheral vein type classification of adolescents/adults/elderly according to their vein characteristics. Initially, authors identified different vein types in literature. This was a preliminary classification that contained the titles and definitions, which were complemented during the expert (nurses, angiologists, anesthetists and biochemists) validation process. Twelve criteria and 27 vein types were validated (agreement level > or =90%). In addition, authors organized a set of pictures representing the different vein types. From these, 144 were used; 35 experts chose one picture to exemplify each classification title. The pictures representing 21 vein types were identified. This classification contributes to the clinical assessment of blood vessels and can be used in teaching, research and patient care management. PMID- 18392537 TI - Performance indicators of DOT at home for tuberculosis control in a large city, SP, Brazil. AB - The study had the objective to analyze the performance of the health services that implement the Directly Observed Therapy at home for tuberculosis control. This study analyzed four Tuberculosis Control Programs, referred to as A, B, C, and D, using the following indicators: Resource use; Performance quickness; Monitoring medication administration; Time spent per home visit. Data were collected during visits to 47 patients receiving DOT at home. Resource use was higher in program B (91.3%); program A showed quicker performance (5.8) and more visits during which medication administration was monitored (77.4%); program C had the longest time spent per home visit (14.7 minutes) and program A the shortest (10.4 minutes). The best or worst performance numerically expresses how resources are being used and whether the observation of medication intake is being achieved. PMID- 18392538 TI - Feelings of women after the diagnosis of type 2 diabetes. AB - Descriptive and exploratory study with the objective of knowing feelings and emotional reactions of women after the diagnosis of diabetes. Data were collected at a primary health care unit in Ribeirao Preto-SP, Brazil, through a semi structured interview. Thematic content analysis was used for interpreting the data. The results showed that women's behavior after the diagnosis of diabetes was associated with feelings and emotional reactions that interfere with treatment adherence and which the health team needs to understand. In an educational process, it is not enough to offer information. Human behavior is extremely complex, going beyond the cognitive aspects, and is rooted in affective emotional dispositions. PMID- 18392539 TI - Study on the absenteeism of nursing professionals in a psychiatric center in Manaus, Brazil. AB - This quantitative study was performed to research the rate of absenteeism of nursing professionals in a psychiatric center in Manaus, from January/2004 to January/2005, in the Human Resources sector of the institution. In this period, the records of workers who were absent from their professional activities for at least one day were checked. Results showed that there were 415 absence reports in the nursing team during the studied period, by 74.29% of nursing professionals. The average rate of absenteeism is 2.79. The main reason reported for absenteeism was illness. These data suggest further studies are needed in order to improve professionals' health, allowing for better quality of life, and, consequently, for providing better health care to Single Health System users. PMID- 18392541 TI - Social representations of the Brazilian national health care system in the city of Rio de Janeiro, Brazil, according to the structural approach. AB - The Brazilian National Health Care System The Single Health System (SHS) [SUS Sistema Unico de Saude] faces difficulties for its effective implementation. This paper aims to characterize the contents and the social representation structure of the Brazilian health care system among health care professionals in the city of Rio de Janeiro. The concept of social memory and the theory of social representation were adopted as frameworks. Five health care institutions were included in this research, with 100 professionals altogether. The free association technique was used to collect data and the EVOC 2003 software was used analyzed for data analysis. The results signal to a central nucleus, characterized by negative attitudes regarding the SUS. In the contrast area, there is a negative attitude towards the effectiveness of the system and a positive attitude towards the care provided to service users, also showing other principles. At its periphery, the implementation of new opinions about the representation could be observed. It is concluded that the professionals present social representations that recognize the SUS as a new system, eliciting negative attitudes among the professionals, and that is in a process of formation or progressive transformation, raising judgments about the pertinence of the system's principles. PMID- 18392540 TI - Illness care at home or in health institutions: the decision process in a low income community. AB - The aim of this research was to describe the experiences of a group of women about the decision making process related to illness care at home or in health care institutions. The thematic oral history method was applied. The descriptive categories were: a) The poor have their own way of taking care of health and illness; b) The main support to fight the illness is belief in God; c) Medical care is the last resource, when healing resources are not enough and the illness is dangerous. In low income families, the cultural background, associated to the socioeconomic conditions, permeate the decision with regard to taking care of the illness at home or to seek institutionalized medical care. This knowledge is essential for meaningful health care from the perspective of individuals and families. PMID- 18392543 TI - Conceptions held by health professionals on violence against children and adolescents within the family. AB - The present study sought to understand the conceptions held by health professionals with regards to violence within the family against children and adolescents. Qualitative case-study methodology and techniques of participant observation, interviewing, and search in documents were used. Participants were staffed in a government-run Family Health Basic Unit in Brazil. Health professionals were found to associate violence with the economic, social, and political juncture and with cultural aspects; for some, violent acts are part of the intergenerational cycle and family dynamics. Physical punishment, considered as violence by some, is advocated as an educational measure by others. Participants also base their definition of violence on an a priori construction of subjects as either victims or aggressors, thus missing the relational dimension of the phenomenon. Health professionals were found to have difficulty in understanding violence in the context that gives it a meaning and to recognize it as consequence of a complex relational dynamics. PMID- 18392542 TI - Opposition to the American leadership by Brazilian nurses (1934-1938). AB - A historical social descriptive study whose objective is to describe the circumstances where Bertha Pullen assumes as the dean of Anna Nery Nursing School, to assess the strategies undertaken by Pullen to assure her position of power and prestige in the space of the school and Brazilian nursing; and to discuss about the resistance to the presence and authority of the American dean for the nurses and students, in the struggle for a national identity. Corpus analysis was made by putting photographs into context based on written documents and secondary sources. RESULTS: The second management of Pullen was the reiteration of the American presence in the leadership of the School, despite the resistance of the nurses and students. Thus, we may assume that the second tenure of Pullen did not represent a real need that was felt by the school. PMID- 18392544 TI - Relationships between parenting styles and risk behaviors in adolescent health: an integrative literature review. AB - Research over the past 20 years suggests that the quality of the parent adolescent relationship significantly affects the development of risk behaviors in adolescent health. The purpose of this paper is to present a review of studies published between 1996-2007 that address specific relationships between parenting styles and six priority adolescent risk behaviors. The review supports the substantial influence of parenting style on adolescent development. Adolescents raised in authoritative households consistently demonstrate higher protective and fewer risk behaviors than adolescents from non-authoritative families. There is also considerable evidence to show that parenting styles and behaviors related to warmth, communication and disciplinary practices predict important mediators, including academic achievement and psychosocial adjustment. Careful examination of parenting style patterns in diverse populations, particularly with respect to physical activity and unintentional injury, will be a critical next step in the development of efficacious, culturally tailored adolescent health promotion interventions. PMID- 18392545 TI - Aluminum as a risk factor for Alzheimer's disease. AB - The purpose of the study was to condense existing scientific evidence about the relation between aluminum (Al) exposure and risk for the development of Alzheimer's Disease (AD), evaluating its long-term effects on the population's health. A systematic literature review was carried out in two databases, MEDLINE and LILACS, between 1990 and 2005, using the uniterms: "Aluminum exposure and Alzheimer Disease" and "Aluminum and risk for Alzheimer Disease". After application of the Relevance Test, 34 studies were selected, among which 68% established a relation between Al and AD, 23.5% were inconclusive and 8.5% did not establish a relation between Al and AD. Results showed that Al is associated to several neurophysiologic processes that are responsible for the characteristic degeneration of AD. In spite of existing polemics all over the world about the role of Al as a risk factor for AD, in recent years, scientific evidence has demonstrated that Al is associated with the development of AD. PMID- 18392546 TI - Collective prenatal consultation: a new proposal for comprehensive health care. AB - This article describes the Collective Prenatal Consultation as a new healthcare methodology, which is performed according to government standards, but collectively. Relaxation and sensitization techniques are used, as well as group dynamics, including a collective exam of the pregnant women. The Collective Consultation is carried out in a welcoming environment, which provides clarification and socialization of experiences and information, centered on those women. The healthcare professional records every obstetric parameter and behavior in the patient's prenatal card and history file. Priority is given to the principle of integrality and citizenship, with the aim to break the biomedical care paradigm, thus favoring humanized and comprehensive care to the women. PMID- 18392548 TI - Insecticide treated mosquito nets for malaria control in India-experience from a tribal area on operational feasibility and uptake. AB - The study assessed the operational feasibility and acceptability of insecticide treated mosquito nets (ITNs) in one Primary Health Centre (PHC) in a falciparum malaria endemic district in the state of Orissa, India, where 74% of the people are tribes and DDT indoor residual spraying had been withdrawn and ITNs introduced by the National Vector Borne Disease Control Programme. To a population of 63,920, 24,442 ITNs were distributed free of charge through 101 treatment centers during July-August 2002. Interview of 1,130, 1,012 and 126 respondents showed that the net use rates were 80%, 74% and 55% in the cold, rainy and summer seasons, respectively. Since using ITNs, 74.5-76.6% of the respondents observed reduction of mosquito bites and 7.2-32.1% reduction of malaria incidence; 37% expressed willingness to buy ITNs if the cost was lower and they were affordable. Up to ten months post-treatment, almost 100% mortality of vector mosquitoes was recorded on unwashed and washed nets (once or twice). Health workers re-treated the nets at the treatment centers eight months after distribution on a cost-recovery basis. The coverage reported by the PHC was only 4.2%, mainly because of unwillingness of the people to pay for re-treatment and to go to the treatment centers from their villages. When the re-treatment was continued at the villages involving personnel from several departments, the coverage improved to about 90%. Interview of 126 respondents showed that among those who got their nets re-treated, 81.4% paid cash for the re-treatment and the remainder were reluctant to pay. Majority of those who paid said that they did so due to the fear that if they did not do so they would lose benefits from other government welfare schemes. The 2nd re-treatment was therefore carried out free of charge nine months after the 1st re-treatment and thus achieved coverage of 70.4%. The study showed community acceptance to use ITNs as they perceived the benefit. Distribution and re-treatment of nets was thus possible through the PHC system, if done free of charge and when personnel from different departments, especially those at village level, were involved. PMID- 18392549 TI - Cadmium accumulation in the edible parts of different cultivars of radish, Raphanus sativus L., and carrot, Daucus carota var. sativa, grown in a Cd contaminated soil. AB - Cadmium accumulation among 12 cultivars of radish (Raphanus sativus) and 10 cultivars of carrot (Daucus carota var. sativa) was studied in a Cd-contaminated soil. The Cd concentration in the edible parts of radish and carrot ranged from 0.04 to 0.14 and 0.14 to 0.19 mg kg(-1) fresh weight, respectively. All the tested carrot cultivars and 33% of the tested radish cultivars exceeded the Chinese allowable limit for Cd. The study showed a greater scope for selecting radish cultivars than for carrot to avoid the excess of the Cd limit when grown on lightly contaminated soils. PMID- 18392550 TI - Is evidence an ethic? PMID- 18392551 TI - Sacral neuromodulation reprogramming: is it an office burden? AB - Our objective was to evaluate the burden of sacral neuromodulation (SNM) reprogramming and the reasons for reprogramming, and to correlate these with the underlying diagnosis. A retrospective review of 50 consecutive subjects implanted with SNM was completed from November 2002 through December 2005. Of the 50 SNM implants, 47 subjects had adequate follow-up. The mean age was 53.1 (22-90 years) with 83% women. Of the SNM performed, 91.5% were done for urinary urgency, frequency, or incontinence and 47.4% of those had interstitial cystitis. Overall, 239 programming visits were recorded, and subjects on average had two reprogramming visits per year. There was no correlation with number of reprogramming events and patient age, sex, reason for implantation, or diagnosis. These data suggest that the number of reprogramming episodes is small, and the majority is done as part of routine follow-up. No additional burden was seen in subjects with interstitial cystitis. PMID- 18392552 TI - The complete nucleotide sequence of an isolate of apple chlorotic leaf spot virus from peach (Prunus persica (L.) Batch). PMID- 18392553 TI - Identification of 13 novel mutations including a retrotransposal insertion in SLC25A13 gene and frequency of 30 mutations found in patients with citrin deficiency. AB - Deficiency of citrin, liver-type mitochondrial aspartate-glutamate carrier, is an autosomal recessive disorder caused by mutations of the SLC25A13 gene on chromosome 7q21.3 and has two phenotypes: neonatal intrahepatic cholestatic hepatitis (NICCD) and adult-onset type II citrullinemia (CTLN2). So far, we have described 19 SLC25A13 mutations. Here, we report 13 novel SLC25A13 mutations (one insertion, two deletion, three splice site, two nonsense, and five missense) in patients with citrin deficiency from Japan, Israel, UK, and Czech Republic. Only R360X was detected in both Japanese and Caucasian. IVS16ins3kb identified in a Japanese CTLN2 family seems to be a retrotransposal insertion, as the inserted sequence (2,667-nt) showed an antisense strand of processed complementary DNA (cDNA) from a gene on chromosome 6 (C6orf68), and the repetitive sequence (17-nt) derived from SLC25A13 was found at both ends of the insert. All together, 30 different mutations found in 334 Japanese, 47 Chinese, 11 Korean, four Vietnamese and seven non-East Asian families have been summarized. In Japan, IVS16ins3kb was relatively frequent in 22 families, in addition to known mutations IVS11 + 1G > A, 851del4, IVS13 + 1G > A, and S225X in 189, 173, 48 and 30 families, respectively; 851del4 and IVS16ins3kb were found in all East Asian patients tested, suggesting that these mutations may have occurred very early in some area of East Asia. PMID- 18392554 TI - Early modification of sickle cell disease clinical course by UDP glucuronosyltransferase 1A1 gene promoter polymorphism. AB - Elevated erythrocyte destruction in sickle cell disease (SCD) results in chronic hyperbilirubinaemia and, in a subset of patients, cholelithiasis occurs. We investigated whether the (TA)n promoter polymorphism in the UDP glucuronosyltransferase 1A1 gene (UGT1A1) may modify bilirubin metabolism, influencing bilirubinaemia, predisposition to cholelithiasis and subsequent cholecystectomy, in a group of 153 young SCD patients (mean age 12.0 +/- 9.0 years) predominantly of Bantu beta S haplotype. The concomitant effect of alpha thalassaemia was also analysed. Among the several UGT1A1 genotypes found, the most frequent were the (TA)6/(TA)6 (n = 37), (TA)6/(TA)7 (n = 60) and (TA)7/(TA)7 (n = 29). These groups of patients did not significantly differ in age, gender ratio and haemoglobin, foetal haemoglobin and reticulocyte levels. On the other hand, total bilirubin levels were significantly different between groups, with an increased (TA) repeat number being associated with higher bilirubinaemia. Furthermore, both cholelithiasis and cholecystectomy were more frequent in groups with higher (TA) repeat number, although the former association was not statistically significant. None of the mentioned parameters is statistically different within UGT1A1 groups with the presence of alpha thalassaemia. Thus, the UGT1A1 promoter polymorphism may represent an important nonglobin genetic modifier of Bantu SCD patients' clinical manifestations, even at a young age. PMID- 18392555 TI - Bumblebee pollination and reproductive biology of Rhododendron semibarbatum (Ericaceae). AB - The reproductive characteristics and pollination system of Rhododendron semibarbatum were investigated at two sites in Honshu, Japan. This species is protandrous, partially self-incompatible at postzygotic stages, and requires outcrossing via pollinator visitation for effective seed production. The effective pollinators were two bumblebee species: males of Bombus ardens at Miyama, and workers of Bombus honshuensis at Agematsu. The flowers possess two staminodes ornamented with whitish hairs, which do not reflect UV light, on the filaments. Nectar was secreted continuously during the flowering period, and nectar production rate differed between the sites. Visitation by B. ardens males was more frequent and varied among and within days, whereas that by B. honshuensis workers was less frequent and constant throughout the observation period. A single visit by a B. ardens male was more effective for seed production than visitation by a B. honshuensis worker, resulting in pollen limitation in the latter case. Differences in resource requirements between the two pollinators, representing different castes, might affect their behavior, resulting in B. ardens males contributing to more effective seed production. PMID- 18392556 TI - Pressure activates colon cancer cell adhesion via paxillin phosphorylation, Crk, Cas, and Rac1. AB - Physical forces can activate colon cancer cell adhesion, critical for metastasis. Paxillin is phosphorylated by FAK and required for pressure-stimulated adhesion. However, whether paxillin acts as an inert scaffolding protein or whether paxillin phosphorylation is required is unknown. Transfection with paxillin point phosphorylation mutants demonstrated that phosphorylation at tyrosines 31 and 118 together is necessary for pressure-stimulated adhesion. We further evaluated potential paxillin partners. Reducing the adaptor protein Crk or the focal adhesion protein p130Cas blocked pressure-stimulated adhesion. Furthermore, Crk and p130Cas both displayed increased co-immunoprecipitation with paxillin in response to increased pressure, except in cells transfected with a Y31Y118 paxillin mutant. Inhibiting the small GTPase Rac1 also abolished pressure stimulated adhesion, and reducing paxillin by siRNA blocked Rac1 phosphorylation by pressure. Thus, paxillin phosphorylation at tyrosines 31 and 118 together is necessary for pressure-induced adhesion. Paxillin, Crk and Cas form a trimeric complex that activates Rac1 and mediates this effect. PMID- 18392557 TI - Warfarin: what are the clinical implications of an out-of-range-therapeutic international normalized ratio? AB - Warfarin is a commonly used oral anticoagulant, and has well-established clinical efficacy. However, it has a narrow therapeutic window, and a mode-of-action affected by inter-individual differences and environmental factors. The effectiveness and safety of warfarin are closely related to maintenance of the international normalized ratio (INR) within therapeutic range. A supra therapeutic INR puts patients at risk of bleeding, whereas a sub-therapeutic INR may not protect against thromboembolic complications. Research suggests a lack of anticoagulation control during warfarin therapy in different settings. Careful monitoring of the INR is essential, especially in geriatric or cancer populations who are at an increased risk of major hemorrhage. Warfarin is an effective treatment but optimization of the risk-benefit ratio is crucial in order to maximize efficacy and safety. Here, we will assess the extent to which INRs are an issue in the management of warfarin therapy, and the effect INRs may have on clinical outcomes. PMID- 18392558 TI - Platelet biology for the clinician-scientist: an evolution of understanding. PMID- 18392560 TI - Organic phase synthesis of ethyl oleate using lipases produced by solid-state fermentation. AB - This paper reports a study of the enzymatic esterification of oleic acid and ethanol. The reaction was catalyzed by lipases produced by solid-state fermentation with Rhizopus sp. Olive oil and perlite were used as an inducer and inert support, respectively. Synthesis of ethyl oleate was carried out in a 10-mL batch reactor with magnetic stirring. The effects of substrate ratios, biocatalyst concentration, and temperature on the reaction rate and conversion efficiency were evaluated. The highest reaction rate (1.64 mmol/L min) was reached with an oleic acid/ethanol mol ratio of 1:5 (oleic acid 50 mM:ethanol 250 mM) and 1 g of biocatalyst. Conversions approaching 100% were obtained after 60 min of reaction at 45 degrees C with n-hexane as a solvent. The initial reaction rate increased proportionally with respect to biocatalyst concentration, which suggests that the reaction rate was not controlled by mass transfer. The biocatalyst retained more than 80% of its catalytic activity after 7 months of storage at 4 degrees C. The results demonstrate that the biocatalyst produced by Rhizopus sp. in solid-state fermentation can be successfully used for ethyl oleate synthesis over short reaction periods under conditions when ethanol is in excess. PMID- 18392559 TI - Microsatellite-based molecular diversity of bread wheat germplasm and association mapping of wheat resistance to the Russian wheat aphid. AB - Genetic diversity of a set of 71 wheat accessions, including 53 biotype 2 Russian wheat aphid (RWA2)-resistant landraces and 18 RWA2 susceptible accessions, was assessed by examining molecular variation at multiple microsatellite (SSR) loci. Fifty-one wheat SSR primer pairs were used, 81 SSR loci were determined, and 545 SSR alleles were detected. These SSR loci covered all the three genomes, 21 chromosomes, and at least 41 of the 42 chromosome arms. Diversity values averaged over SSR loci were high with mean number of SSR alleles/locus = 6.7, mean Shannon's index (H) = 1.291, and mean Nei's gene diversity (He) = 0.609. The three wheat genomes ranked as A > D > B and the homoeologous groups ranked as 7 > 3 > 1 > 2 > 6 > 5 > 4 based on the number of alleles per locus. Xgwm136 on chromosome arm 1AS is the most polymorphic SSR locus with the largest number of observed and effective alleles and the highest H and He. Among all 2485 pairs of wheat accessions, genetic distance (GD) ranged from 0.054 to 1.933 and averaged 0.9832. A dendrogram based on GD matrix showed that all the wheat accessions could be grouped into distinct clusters. Most of the susceptible cultivars (13/18) were clustered into groups that contains all or mostly susceptible accessions. Most of the U.S. cultivars belong to a group that is distinguishable from all the different RWA2 resistant groups. Diversity analysis was also conducted separately for subgroups containing 53 RWA2-resistant accessions and 18 RWA2-susceptible accessions. Association mapping revealed 28 SSR loci significantly associated with leaf chlorosis, and 8 with leaf rolling. New chromosome regions associated with RWA2 resistance were detected, and indicated existence of new RWA resistance genes located on chromosomes of all other homoeologous groups in addition to the groups 1 and 7 in bread wheat. This information is helpful for development of mapping populations for RWA2 resistance genes from different phylogenetic groups, and for wise utilization of the RWA resistant germplasm in wheat breeding programs. PMID- 18392563 TI - Determination of small ions with capillary electrophoresis and contactless conductivity detection. AB - Capillary Electrophoresis (CE) has become an accepted method for the separation of inorganic and organic ions. Usually, direct and indirect optical detection methods are used in conventional CE. However, with contactless conductivity detection, much better detection limits in the low ppb range are obtained compared to optical detection modes. Besides offering great flexibility in capillary handling, this detection technique can be performed on-capillary also with capillaries made of other materials than fused silica (PEEK, Teflon) and with capillaries having very small inner diameters. PMID- 18392561 TI - Production of biopesticides in an in situ cell retention bioreactor. AB - The seeds of Azadirachta indica contain azadirachtin and other limonoids, which can be used as a biopesticide for crop protection. Significant variability and availability of seed only in arid zones has triggered biotechnological production of biopesticides to cope up with its huge requirement. Batch cultivation of A. indica suspension culture was carried out in statistically optimized media (25.0 g/l glucose, 5.7 g/l nitrate, 0.094 g/l phosphate and 5 g/l inoculum) in 3 l stirred tank bioreactor. This resulted in 15.5 g/l biomass and 0.05 g/l azadirachtin production in 10 days leading to productivity of 5 mg l(-1) day(-1). Possible inhibition by the limiting substrates (C, N, P) were also studied and maximum inhibitory concentrations identified. The batch kinetic/inhibitory data were then used to develop and identify an unstructured mathematical model. The batch model was extrapolated to simulate continuous cultivation with and without cell retention in the bioreactor. Several offline computer simulations were done to identify right nutrient feeding strategies (with respect to key limiting substrates; carbon, nitrate and phosphate) to maintain non-limiting and non inhibitory substrate concentrations in bioreactor. One such continuous culture (with cell retention) simulation was experimentally implemented. In this cultivation, the cells were propagated batch-wise for 8 days. It was then converted to continuous cultivation by feeding MS salts with glucose (75 g/l), nitrate (10 g/l), and phosphate (0.5 g/l) at a feed rate of 500 ml/day and withdrawing the spent medium at the same rate. The above continuous cultivation (with cell retention) demonstrated an improvement in cell growth to 95.8 g/l and intracellular accumulation of 0.38 g/l azadirachtin in 40 days leading to an overall productivity of 9.5 mg l(-1) day(-1). PMID- 18392564 TI - Metal analysis with capillary zone electrophoresis. AB - Capillary electrophoresis has recently attracted considerable attention as a promising analytical technique for metal ion separations. Significant advances in various auxiliary separation principles have opened new application areas for capillary electrophoresis in the analysis of metal species. These advances are mainly due to complexation, ion pairing, solvation and micellization interactions between metal analytes and electrolyte additives, which alter the separation selectivity in a broad range. Likewise, many separation studies on metal ions have been concentrated on the use of pre-electrophoresis derivatization methodology. Approaches suitable for improvement of selectivity for different metal species including metal cations, metal complexes, metal oxoanions and organometallic compounds are discussed, with special attention paid to the related electrophoretic system variables using illustrative examples. PMID- 18392565 TI - Measurement of low-molecular-weight carboxylic acids in ambient air and vehicle emission by capillary electrophoresis. AB - Within the last few years, capillary electrophoresis (CE), especially with indirect ultraviolet detection, has successfully been utilized for the analysis of low-molecular-weight (LMW) organic acids in a wide variety of matrices (e.g., food, pharmaceutical, environmental, industrial, clinical). The speed, resolution, and simplicity of CE, combined with low operating costs, make the technique an attractive option for the development of improved methods in this field. Hence, CE is becoming increasingly accepted for routine analytical work. In this chapter, the unique capability and applicability of the five selected CE methods used in the analysis of LMW carboxylic acids in ambient air and/or vehicle-emitted samples are described. PMID- 18392566 TI - Determination of aliphatic low-molecular-weight and biogenic amines by capillary zone electrophoresis. AB - Low-molecular-weight (LMW) aliphatic amines play a key role in the global nitrogen cycle, are involved in nutrient transfer, and act as buffer in the ecosystem. They are widely used as intermediates in chemical synthesis and were shown to cause occupational asthma. Biogenic amines occur in all living organisms and have an effect on the cell growth, although at high concentrations they can be toxic; some are used as cancer markers in health protection or as spoilage markers in foods. Their identification and quantification from different matrices such as human tissues or foods is of high importance. The electrophoretic separation of amines is possible as cations as a result of their high basicity; their detection, however, is more difficult because these amines contain no chromophore group. Indirect ultraviolet (UV) detection is the first presented possibility and widely used for the separation of nonderivatized amines. Otherwise, derivatization of the amines is necessary to directly detect them with laser-induced fluorescence (LIF) detection. Other detection modes such as pulse amperometric, chemiluminescence, or mass spectrometry have been also used for the determination of LMW and biogenic amines, but not on a routine basis. In this chapter, three capillary electrophoretic methods with indirect UV and LIF detection for determination of LMW aliphatic and biogenic amines are described. PMID- 18392567 TI - Capillary electrophoretic analysis of organic pollutants. AB - Environmental pollutants comprise a variety of compounds from inorganic anions, cations, ionizable organic compounds and moderately hydrophobic organic compounds to highly hydrophobic organic compounds. Correspondingly different separation strategies are required for their separation. In this chapter, we have presented some methods for the separation and the analysis of the organic pollutants such as polycyclic aromatic hydrocarbons, phenoxy acids, dithiocarbamates, paraquat and diquat, endocrine disruptors, toxins and explosives. PMID- 18392568 TI - Capillary electrophoresis with laser-induced fluorescence: environmental applications. AB - Capillary electrophoresis (CE), especially free-zone CE, offers a relatively simple separation with moderate selectivity based on the mobility of ions in solution. Laser-induced fluorescence (LIF) detection, an extremely sensitive technique, can be coupled with a variety of separation conditions to achieve sensitive and quantitative results. When these techniques are combined, CE/LIF provides the sensitivity and increased selectivity that makes trace level environmental analysis of fluorescent compounds possible at or below levels typical for gas chromatography (GC)/mass spectrometry (MS). We offer a panoramic review of the role of these tools in solving environmental and related analytical problems before providing a detailed experimental protocol. PMID- 18392569 TI - Practical considerations for the analysis of ionic and neutral organic molecules with capillary electrophoresis/mass spectrometry. AB - This chapter presents the technique of capillary electrophoresis coupled to mass spectrometry (CE/MS). The introductory section is targeted mainly at CE/MS beginners and notes briefly the theoretical background of electrospray ionization (ESI), the most commonly used ionization mode in CE/MS. The specifics of CE/MS are described--also in comparison with more classic methods like LC/MS. Important caveats to be taken into consideration for successful CE/MS operation are noted in the interest of avoiding pitfalls. CE/MS is illustrated with three representative examples, which might serve as starting points for more in-detail experiments: (1) partial-filling micellar electrokinetic chromatography (MEKC) of neutral bacterial signaling molecules (N-acylhomoserine lactones) extracted from culture supernatants, (2) capillary zone electrophoresis (CZE) of their anionic degradation products, and finally (3) CZE separation of cationic hydroxy-s triazines. PMID- 18392570 TI - Analysis of the enantiomers of chiral pesticides and other pollutants in environmental samples by capillary electrophoresis. AB - The generic method described here involves typical capillary electrophoresis (CE) techniques, with the addition of cyclodextrin chiral selectors to the electrolyte for enantiomer separation and also, in the case of neutral analytes, the further addition of a micelle-forming compound such as sodium dodecyl sulfate (SDS) for separation by the micellar electrokinetic chromatography (MEKC) mode of CE. This generic method has broad application for the separation and analysis of enantiomers of chiral pesticides and other small molecules in a variety of environmental matrices. Aqueous samples such as surface water are analyzed after simple filtration, but centrifugation is sometimes necessary for soil-water slurry samples. Soils and sediment must be extracted with a polar organic solvent such as methanol, which needs only to be evaporated to near dryness, diluted with water, and filtered before CE analysis. Simple borate or phosphate-based buffers are usually used in the CE electrolyte. The method must be optimized for the electrolyte composition, including the correct chiral selector and its concentration, as well as for column conditions and instrumental variables such as voltage. Specific methodologies for application of this generic CE method to follow the enantioselective microbial transformation of ruelene, a neutral organophosphorus insecticide, dichlorprop, an ionic phenoxyalkanoic acid herbicide, and bromochloroacetic acid, a drinking water disinfection byproduct, are provided. PMID- 18392571 TI - Capillary electrophoresis of tropane alkaloids and glycoalkaloids occurring in Solanaceae plants. AB - This chapter examines the role of capillary electrophoresis (CE) in the separation of tropane alkaloids, glycoalkaloids, and closely related compounds that have either pharmaceutical value or toxicological effects on humans. The latest significant developments in CE analysis have been selected and critically discussed. When the conventional CE mode was found unable to provide an acceptable selectivity towards the analytes, the addition of either an organic solvent, a chiral selector, or a surfactant to the running buffers was exploited. Likewise, nonaqueous CE (NACE) was also employed to increase solute solubilities and for a better compatibility of this media with mass spectrometry. It turns out that, upon selecting the most appropriate experimental conditions, the CE separation of tropane alkaloids and steroidal glycoalkaloids of Solanaceae plants was successfully accomplished. All major steps involved in the separation and detection of these secondary metabolites in complex samples are described and the relevant aspects of each application are examined with emphasis on the main aspects entailed a typical assay. More applications have yet to be developed in order to encourage more labs to exploit the tremendous potential of capillary electrophoresis. PMID- 18392572 TI - Capillary electrophoresis for pharmaceutical analysis. AB - This chapter describes the application of capillary electrophoresis (CE) to pharmaceutical analysis. The areas of pharmaceutical analysis covered are enantiomer separation, analysis of small molecules such as amino acids or drug counter-ions, pharmaceutical assay, related substances determinations, and physiochemical measurements such as log P and pKa of compounds. The different electrophoretic modes available and their advantages for pharmaceutical analysis are described. Recent applications of CE for each subject area are tabulated with electrolyte details. Information on electrolyte choice and method optimization to obtain optimal separations is included. PMID- 18392573 TI - Capillary electrophoresis of neutral carbohydrates: mono-, oligosaccharides, glycosides. AB - This chapter reports an overview of the recent advances in the analysis of neutral sugars by capillary electrophoresis (CE); furthermore, some relevant reviews and research articles in the field are tabulated. Comparison of CE with chromatography is also presented, with special attention to separation efficiency and sensitivity. The main routes aimed at pretreatment and CE analysis of uncharged mono-, oligosaccharides, and glycosides are described. Representative examples of such procedures are reported in detail, upon describing robust methodologies for the study of (1) neutral mono- and oligosaccharides derivatized by reductive amination and by formation of glycosylamines; (2) underivatized mono and di-saccharides analyzed using highly alkaline buffers; and (3) anomeric couples of glycosides separated using borate-based buffers. PMID- 18392574 TI - Capillary electrophoresis of sugar acids. AB - This chapter illustrates the usefulness of capillary electrophoresis (CE) for the analysis of sugar acids, that is, monosaccharides and lower oligosaccharides carrying carboxylate, sulphate or phosphate groups. In order to provide a general description of the main results and challenges in the field, some relevant applications and reviews on CE of such saccharidic compounds are tabulated. Furthermore, some detailed experimental procedures are shown, regarding the CE analysis of sugar acids released upon hydrolysis of acidic polysaccharides and of glycans linked to glycoproteins. In particular, the protocols will deal with the following compounds: (i) unsaturated, underivatized oligosaccharides from lyase treated alginate; (ii) oligosaccharides derivatized with 4-aminobenzonitrile, arising from chemical hydrolysis of alginate; (iii) sialic acid derivatized with 2-aminoacridone, released from human serum immunoglobulin G. PMID- 18392575 TI - Use of capillary electrophoresis for polysaccharide studies and applications. AB - Capillary electrophoresis (CE) applications to charged polysaccharides are briefly reported. A simple procedure is presented to determine the esterification degree of a hyaluronan derivative. In this case, the degree of substitution was as low as 14%. The molecular weight distribution of mannuronic oligosaccharides mixture produced by hydrolysis of native polymannuronic is readily calculated from peak area of the species resolved by CE on the basis of a specific degree of polymerization. The influence of the applied electric field strength on the free solution mobility of hyaluronan samples is briefly addressed for molar masses of the order of 10(5) and 10(6) g/mol. The data are compared with the results obtained for a 50% galactose-substituted hyaluronic acid (HA). Mobility data obtained as a function of buffer pH for a native HA sample as well as for two galactose-amide HA derivatives, having slightly different degrees of substitution, are presented and discussed in terms of the polymer charge density parameters xi. In most cases, more questions than answers arise from the application of CE to charged polysaccharides. However, perspectives are disclosed for a further understanding of the reliability of CE applied for the structural elucidation of such macromolecules. PMID- 18392577 TI - Separation of DNA by capillary electrophoresis. AB - This chapter reports an overview of the analytical techniques used to perform genetic analysis of polymerase chain reaction products using capillary electrophoresis. Three separate but related techniques are described: the separation of native DNA with detection using fluorescent intercalating dyes, the separation of denatured DNA using fluorescently labeled primers, and the detection of single-strand conformation polymorphisms using denatured DNA separated under native conditions. The various techniques involve electrokinetic injection of the DNA onto a narrow band at the head of the column, sieving the DNA through various entangled polymer matrices, and detection via single or multichannel laser-induced fluorescence. Analytical protocols are provided, and a series of representative electropherograms are included. PMID- 18392576 TI - Analysis of oligonucleotides using capillary zone electrophoresis and electrospray mass spectrometry. AB - This chapter illustrates the usefulness of capillary zone electrophoresis (CZE) coupled to high-resolution electrospray ionization quadrupole time-of-flight mass spectrometry for the single-step desalting, and separation, as well as characterization of oligonucleotides in the framework of quality control after synthesis. Separation is performed using a 25 mM ammonium carbonate buffer supplemented with 0.2 mM trans-1,2-diaminocyclohexane-N, N, N', N' id (CDTA) (pH 9.7). During the electrophoretic process, sodium and potassium ions are removed from the polyanionic backbone of the oligonucleotides by exchange of these ions with ammonium ions or by chelation on CDTA, thus eliminating a sample preparation step. A sample stacking procedure used to concentrate the samples on the CZE capillary is described. After analysis, the obtained spectrum is deconvoluted to the zero charge spectrum to yield the molecular mass of the oligonucleotide. A misincorporation of one nucleotide can be detected by a difference in mass. PMID- 18392578 TI - Capillary electrophoresis of oxidative DNA damage. AB - Urinary 8-hydroxy-2'-deoxyguanosine (8OHdG) is an excellent marker of oxidative DNA damage. Until now, urinary 8OHdG has been measured by high-performance liquid chromatography with electrochemical detection. A simple and sensitive method for the analysis of urinary 8OHdG by capillary electrophoresis with end-column amperometric detection has been developed and is described in this chapter. A single-step solid-phase extraction procedure was optimized and used for extracting 8OHdG from human urine. To improve the sensitivity of this method, a new focusing technique based on a dynamic pH junction was used. In the end, the urinary concentration of 8OHdG in healthy persons, patients with cancer, patients with diabetic nephropathy, and smokers was determined. Emphasis is focused on the establishment and application of the methods. PMID- 18392579 TI - Capillary electrophoresis of gene mutation. AB - This chapter illustrates the usefulness of capillary electrophoresis (CE) for the detection of gene mutation, i.e., point mutation, methylation, and microsatellite analysis. In order to provide a general description of the main results and challenges in the field, some relevant applications and reviews on CE of gene mutation are tabulated. Furthermore, some detailed experimental procedures are shown. Several CE methods of gene mutation detection were developed including the following: (1) single-strand conformation polymorphism with capillary electrophoresis; (2) SNaPshot analysis; (3) constant denaturant capillary electrophoresis; (4) microsatellite analysis; and (5) methylation analysis. PMID- 18392581 TI - Separation of peptides by capillary electrophoresis. AB - Peptides are an important class of analytes in chemistry, biochemistry, and food chemistry as well as medical and pharmaceutical sciences. As a high-resolution technique, capillary electrophoresis (CE) is well suited for the analysis of polar compounds such as peptides. In addition, CE is orthogonal to high performance liquid chromatography, as both techniques are based on different physico-chemical separation principles. For the successful development of peptide separations by CE, operational parameters including buffer pH, buffer concentration and buffer type, applied voltage, and capillary dimensions, as well as background electrolyte additives such as detergents, ion-pairing reagents, cyclodextrins, (poly)amines, soluble polymers, etc. must be considered and optimized. PMID- 18392580 TI - Biomedical applications of amino acid detection by capillary electrophoresis. AB - Capillary electrophoresis (CE) is an efficient tool for amino acid (AA) analysis. However, its role can be fully accounted for only by examining the applications on real matrices. Methods must be successfully transferred into working environments for use by non-CE experts before their power can be realized. This transfer of technology is rapidly increasing. In this chapter, some applications to real samples are presented with the precise intent to illustrate the great capabilities of CE to AA analysis in clinical applications. PMID- 18392582 TI - Analysis of proteins by capillary electrophoresis. AB - This chapter describes the basic principles of protein analysis by capillary electrophoresis, and provides an overview of the literature and thus a comprehensive summary of special topics in this field. The "Materials and Methods" section includes the main experimental points to be taken into consideration, namely, sample pretreatment, reduction of protein adsorption to capillary wall, increase of selectivity, detection modes, and special electrophoretic modes. Because there are many capillary electrophoretic methods for protein analysis, not all experimental steps are listed; however, the main references are cited. Additional experimental information can be found in the Notes. Finally, an overview of the most relevant applications, divided according to the origin of the samples into human proteins, food and agricultural products, pharmaceutical proteins, proteome, and special proteins, is given with the relevant literature. PMID- 18392583 TI - Separation of synthetic (co)polymers by capillary electrophoresis techniques. AB - Capillary electrophoresis (CE) is a very efficient tool for separating and characterizing synthetic polymers, copolymers, and polyelectrolytes. Different modes of CE (free solution capillary electrophoresis [FSCE], entangled polymer solution CE [EPSCE], capillary gel electrophoresis [CGE], or micellar electrokinetic chromatography [MEKC]) can be used depending on the characteristics of the polymer solutes (end charged, evenly charged, or uncharged polymers) and on the polymer solute heterogeneities (molecular mass, functionality, chemical composition). To illustrate the potential of CE, four different methods are proposed using either nonaqueous or aqueous electrolytes. The first method describes the separation of synthetic organic polypeptides according to their functionalities and molar masses in a nonaqueous electrolyte. In a second method, polyelectrolyte oligomers are separated by FSCE in aqueous buffer. The third method demonstrates the great potential of EPSCE for the size based separation of evenly charged polyelectrolytes on a wide range of molar masses. The last method describes a simple two-dimensional approach realized in a single capillary that combines a separation according to the chemical composition (FSCE) with a size-based separation (EPSCE). PMID- 18392584 TI - Capillary electrophoresis separation of microorganisms. AB - Microorganisms can be considered a bio-colloid. That is, they have a characteristic outer surface that carries, or can carry, a charge. Precisely, differences in the surface can be exploited for separation by capillary electrophoresis (CE). In fact, methods based on CE seem to be very promising because they should produce rapid and high-efficiency separations. Although CE can be used to separate microbial (i.e., bacteria, virus, fungi, and whole cells) and subcellular particles (i.e., mitochondria and nuclei), this chapter is focused mainly on the determination of bacteria and virus for their interest. At difference to the separation off molecules, microorganisms are characterized as living. This makes their analysis more difficult because several aspects such as possible lysis, aggregation, evolution, growing etc. must be taken into count. PMID- 18392585 TI - A semi-empirical approach for a rapid comprehensive evaluation of the electrophoretic behaviors of small molecules in free-zone electrophoresis. AB - A phenomenological model is proposed for the evaluation of relative electrophoretic migration of charged substances present in mixtures and for the rapid pH optimization prior to capillary zone electrophoresis method development. The simple and robust model is based on the Offord model, which takes account of the chemical structure. The effective charge and the molecular mass of the molecule are needed; the charge can easily be calculated from pKa obtained from known sources or simulated with existing pK-calculation programs. A first example was chosen with the separation of hydroxy-s-triazines to illustrate the applicability of this simple approach for determination of the first buffer-pH conditions prior experimental method optimization when separation of different ions is needed. In a second example, the confirmation of aminialcohols in the CZE method development of unsaturated hexahydro-triazines and oxasolidines. PMID- 18392586 TI - The CE way of thinking: "all is relative!". AB - Over the last two decades, the development of capillary electrophoresis (CE) instruments has lead to systems with programmable samplers, separation columns, separation buffers, and detection devices comparable visually in many aspects to the setup of classical chromatography. Two characteristics make CE essentially different from chromatography and are the basis of the CE way of thinking: first is the injection type and the liquid flow within the capillary. When the injection is made hydrodynamically (such as in most of the applications found in the literature), the injected volumes are directly dependent on the type and size of the separation capillary. The second characteristic is that in CE, buffer velocity is not pressure-driven, as in liquid chromatography, but is electrokinetically governed by the quality of the capillary surface (separation buffer dependent surface charge) inducing an electroosmotic flow (EOF). The EOF undergoes small variations and is not necessarily identical from one separation or day to the other. The direct consequence is that the migration time of the analytes apparently nonreproducible, although the velocity of the ions is the same. The effective mobility (field strength normalized velocity) of the ions is a possible parameterization from acquired time-scale to effective mobility-scale electropherograms leading to a reproducible visualization and better quantification with a direct relation to structural characters of the analytes (i.e., charge and size; see Chapter 23). PMID- 18392587 TI - Adsorbed cationic polymer coatings for enhanced capillary electrophoresis/mass spectrometry of proteins. AB - The combination of capillary electrophoresis (CE) with mass spectrometry (MS) constitutes a powerful microanalytical system for the analysis of biological samples. The anionic and hydrophobic surface of the fused-silica capillary is, however, known to cause severe analyte-wall interactions in protein analysis. In order to control surface properties and eliminate protein adsorption, a capillary coating can be applied. A fast and simple strategy is to coat the anionic capillary with a cationic polymer via multisite electrostatic interaction. This generates a stable deactivation layer, without the need for addition of coating agent to the background electrolyte solution. This chapter reviews the present knowledge of capillary coatings and especially cationic polymers in CE-MS, and describes the synthesis of a cationic polymer, PolyE-323, for deactivation of fused-silica capillaries. The capillary coating procedure is a simple three-step rinsing protocol comprising deprotonation of surface silanol groups using a base, adsorption of polymer, and a final rinse to remove excess polymer not adsorbed to the surface. As a result of the simplicity of the coating procedure, highly reproducible coatings can be prepared with little or no expert skills. Some practical aspects on using cationic-coated capillaries in CE-MS protein analysis are also discussed. PMID- 18392589 TI - On-line concentration of environmental pollutant samples by using capillary electrophoresis. AB - This chapter reviews the theory and methodological developments of on-line concentration techniques for the determination of environmental pollutant samples, such as organic and inorganic compounds in capillary zone electrophoresis (CZE) and also in micellar electrokinetic chromatography (MEKC). Topics covered include a variety of on-line preconcentration strategies, which are now generally referred to as sample stacking and sweeping techniques. For each technique, surveyed methods are tabulated in order to assist in method selection. Innovative applications of sample stacking and sweeping to advanced environmental research are also emphasized. In addition, other comparative on line concentration methods for environmental samples, namely, isotachophoretic stacking and anion and cation selective exhaustive injection-sweeping are briefly discussed. PMID- 18392588 TI - On-column ligand/receptor derivatization coupled to affinity capillary electrophoresis. AB - The coupling of on-column derivatization of small molecules to affinity capillary electrophoresis (ACE) has only been realized during the past 5 yr. In this technique, multiple zones of reagent(s) and ligand or receptor are injected into the capillary column. Upon electrophoresis, zones of sample overlap, yielding product. Continued electrophoresis results in the product overlapping with receptor (or ligand, if the receptor was derivatized), thereby causing a shift in migration time of the compound in question. Subsequent Scatchard analysis using noninteracting standards realizes a binding constant. Herein, we describe the use of on-column-ligand and receptor derivatization coupled to partial-filling ACE (PFACE) to probe the binding of vancomycin (Van) from Streptomyces orientalis and teicoplanin (Teic) from Actinoplanes teicomyceticus to D-Ala-D-Ala terminus peptides. PMID- 18392590 TI - Free-flow electrophoresis system for proteomics applications. AB - This chapter describes the technology and selected applications in the field of protein analysis for free flow electrophoresis (FFE). FFE is a highly versatile technology for applications in this field as a result of its continuous processing of sample and the possibility of separating almost any kind of charged or chargeable particles in an aqueous medium without any solid matrix such as acrylamide. Possible applications are separation of ions, proteins, peptides, organelles, or whole cells for subsequent analysis. PMID- 18392591 TI - Microemulsion electrokinetic chromatography. AB - Microemulsion electrokinetic chromatography (MEEKC) is an attractive capillary electrophoretic technique in which a microemulsion is used as carrier electrolyte. Analytes may partition between the aqueous phase of the microemulsion and its oil droplets, which act as a pseudostationary phase. It is well suited for the separation of neutral analytes, but can also be employed for charged analytes. A single set of separation parameters may be sufficient for separation of a wide range of analytes. Fine-tuning of the separation may be achieved by addition of organic solvents to the microemulsion or by changes in the nature of the surfactant used for stabilization of the microemulsion. In this chapter, MEEKC conditions are summarized that have proven their reliability for routine purposes. Furthermore, microemulsions can be used for on-capillary preconcentration of analytes so that the problem of poor concentration sensitivity of ultraviolet detection in capillary electrophoresis is circumvented. PMID- 18392592 TI - Micellar electrokinetic chromatography of aminoglycosides. AB - The components of the aminoglycosides, e.g., gentamicin, sisomicin, netilmicin, kanamycin, amikacin, and tobramycin, and related impurities of these antibiotics can be separated by means of micellar electrokinetic chromatography (MEKC). Derivatization with o-phthaldialdehyde and thioglycolic acid is found to be appropriate for all antibiotics. The background electrolyte was composed of sodium tetraborate (100 mM), sodium deoxycholate (20 mM), and beta-cyclodextrin (15 mM) and has a pH value of 10.0. This method is valid for evaluation of gentamicin, kanamycin, and tobramycin. It has yet to be adopted for amikacin, paramomycin, neomycin, and netilmicin. PMID- 18392593 TI - Capillary electrochromatography and on-line concentration. AB - Capillary electrochromatography (CEC) is a micro-separation technique that combines the advantages of capillary zone electrophoresis with those of high performance liquid chromatography. Accordingly, it has attracted extensive attention over the last decade. Among the stationary phases for CEC, monolithic stationary phase has been regarded as the most suitable stationary phase for CEC because of its simple preparation, the elimination of frits, and its excellent performance. In this chapter, procedures for preparing CEC monolithic columns with an improved configuration, in which there are stationary phases at both sides of detection window and no stationary phase at detection window, are presented. The separation of acidic and basic compounds on such monolithic columns is used as an example to demonstrate CEC separation protocol. Additionally, an on-line concentration technique in CEC is presented. As a result of the coexistence of stationary phase and electric field in a CEC column, it is possible to employ chromatographic zone sharpening and field-amplified sample stacking effects simultaneously to improve CEC detection sensitivity. PMID- 18392594 TI - Analysis of alkaloids in single plant cells by capillary electrophoresis. AB - In this chapter, capillary electrophoresis (CE) is demonstrated to be a useful technique for the determination of alkaloids in microsamples of single plant cells. A single cell sampling technique with microcapillaries that includes extraction of sample volumes in the pl range from single cells, division into aliquots, addition of internal standard, and injection into the CE capillary is described. The danger of contamination and evaporation of such low sample volumes has been avoided by handling them under an inert protective layer of silicone oil. For the determination of alkaloids in cell samples, CE with direct ultraviolet detection using a high concentration of citric acid as background electrolyte provides sufficient sensitivity. PMID- 18392596 TI - Knotless anatomic double-layer double-row rotator cuff repair: a novel technique re-establishing footprint and shape of full-thickness tears. AB - The standard technique for restoring footprint after full-thickness tears of the rotator cuff includes double-row or transosseous-equivalent techniques. However, the anatomically typical bird's beak shape and profile of tendon insertion may not be originally restored and biomechanics may be altered. In this report, the authors describe a technique that involves creating two intratendinous stitches at different levels of the torn tendon. The first passes through the bursal-side layer, the second stitch through the joint-side layer. Both stitches may be performed in mattress suture configuration. The anchorage is performed by knotless anchors in order to avoid knots lying within the insertion area. The footprint is restored first medially then laterally by the use of double-row principles. The joint-side suture is anchored within the medially placed anchor. The bursal-side suture is anchored by a laterally placed anchor. The anatomic insertion and restoration of the shape and profile may be enabled by the described double-layer suture technique. Using a double-layer double-row repair may potentially improve functional results of rotator cuff repair constructs. PMID- 18392595 TI - Multi-dimensional capillary electrophoresis and chromatography for proteomic analysis. AB - Comprehensive two-dimensional liquid chromatography-capillary electrophoresis systems are summarized in this chapter. A variety of combinations of capillary electrophoresis and liquid chromatography modes as well as interfaces and detection technologies are discussed. A typical, comprehensive two-dimensional system coupled with reverse-phase liquid chromatography with fast capillary electrophoresis and hyphenated to mass spectrometry was demonstrated for proteomic analysis. A two-dimensional capillary electrophoresis system of coupling capillary sieving electrophoresis with micellar electrokinetic chromatography and its application in single cell analysis for protein expression profiling are presented. PMID- 18392597 TI - Spontaneous resolution of solitary rectal ulcer syndrome during pregnancy: report of a case. AB - Solitary rectal ulcer syndrome is a rare condition that is associated with disordered defecation and is thought to be an ischemic injury from repeated mucosal trauma. Treatment is designed to alleviate the underlying defecatory problems and is only moderately successful. We report an interesting case of solitary rectal ulcer syndrome in a young woman, which was resistant to standard nonoperative management but completely resolved during two pregnancies only to recur when she was not pregnant, and we suggest a possible hormonal explanation for this unique occurrence. PMID- 18392598 TI - Secreted frizzled related protein 1 (Sfrp1) and Wnt signaling in innervated and denervated skeletal muscle. AB - Wnts are secreted proteins with functions in differentiation, development and cell proliferation. Wnt signaling has also been implicated in neuromuscular junction formation and may function in synaptic plasticity in the adult as well. Secreted frizzled-related proteins (Sfrps) such as Sfrp1 can function as inhibitors of Wnt signaling. In the present study a potential role of Wnt signaling in denervation was examined by comparing the expression levels of Sfrp1 and key proteins in the canonical Wnt pathway, Dishevelled, glycogen synthase kinase 3beta and beta-catenin, in innervated and denervated rodent skeletal muscle. Sfrp1 mRNA and immunoreactivity were found to be up-regulated in mouse hemidiaphragm muscle following denervation. Immunoreactivity, detected by Western blots, and mRNA, detected by Northern blots, were both expressed in extrasynaptic as well as perisynaptic parts of the denervated muscle. Immunoreactivity on tissue sections was, however, found to be concentrated postsynaptically at neuromuscular junctions. Using beta-catenin levels as a readout for canonical Wnt signaling no evidence for decreased canonical Wnt signaling was obtained in denervated muscle. A role for Sfrp1 in denervated muscle, other than interfering with canonical Wnt signaling, is discussed. PMID- 18392599 TI - [The economics of contralateral laparoscopic inguinal hernia exploration. Cost calculation of herniotomy in infants]. AB - BACKGROUND: Diagnostic laparoscopy (DL) of the contralateral side during inguinal herniotomy via the hernia sack may avoid a subsequent second operation. Can this procedure however also reduce costs in the German health care system? METHODS: Prospective analysis was performed of children operated on for inguinal hernia (IH) from March 2006 until October 2007. Using a linear mathematic model, the costs for different scenarios were calculated regarding the risk of contralateral IH. We thereby determined the incidence of contralateral IH at which DL became economically reasonable. RESULTS: A total of 123 unilateral IH operations (IH-OP) were performed in infants during the study period. Of these, 31 patients underwent DL of the contralateral side. Thirteen open hernia sacks were identified and ligated during the same operation. The following costs were calculated: (1) IH-OP without DL, 286 Euro, (2) IH-OP with contralateral DL, 338 Euro, (3) IH-OP with DL and synchronous ligation of the contralateral side, 393 Euro, and (4) metachronous operations of bilateral IH, 572 Euro. The incidence of contralateral hernia described in the literature ranges from 20% to 50%. Linear regression of the relative costs shows an economic advantage for DL with an incidence above 23%. CONCLUSION: Laparoscopic evaluation of the contralateral side in IH-OP is a rational approach for the patient and makes economic sense in the German health care system. PMID- 18392600 TI - [Screening: prerequisites]. AB - Screening can be a very powerful tool for prevention or more effective treatment of diseases. However, a number of prerequisites have to be met. Only diseases with a preclinical phase, during which the disease or its precursors can be detected by a suited test, are amenable to screening. Early detection of the disease must enable either prevention or more effective management of the disease and not just prolong the "patient career". The benefits of screening must encompass potential harms, which may include, for example, complications, false positive diagnoses or over-diagnoses (i.e. the diagnosis of clinically irrelevant disease). Benefits from screening must be achieved at acceptable costs. Implementation of screening has to be based on scientific evidence and accompanied by scientific evaluation. PMID- 18392601 TI - Young fire ant workers feign death and survive aggressive neighbors. AB - Feigning death is a method of self-defense employed among a wide range of prey species when threatened by predator species. This paper reports on death-feigning behavior by the fire ant, Solenopsis invicta, during intraspecific aggression among neighboring fire ant workers. Days-old workers responded to aggression by death feigning, weeks-old workers responded by fleeing and months-old workers responded by fighting back. By feigning death, days-old workers were four times more likely to survive aggression than older workers. From a proximate perspective, retaliation by young workers against aggressive older workers is certain to fail. With their relatively soft exoskeleton, young workers would be prone to injury and death and unable to execute an effective attack of biting or stinging older workers with harder exoskeletons. From an ultimate perspective, death feigning allows young workers to survive and contribute to brood care and colony growth, both of which are essential to queen survival and fitness. PMID- 18392602 TI - [Acute spinal cord ischemia following deep injection therapy of the neck]. AB - We report the case of a 40-year-old woman suffering from neck pain due to mild cervical spine injury. During conventional faceted infiltration therapy with crystalline steroids, the patient developed weakness in both arms and paresthesia of the left arm. While the weakness resolved within a few seconds, a mild deficit in motor coordination and paresthesia of the left arm were still present after 2 months. T2-weighted magnetic resonance imaging of the cervical spine depicted a small hyperintense lesion at the C6 level most likely reflecting embolic infarction due to injection of crystalline steroids into a myelon-feeding artery, which has of late repeatedly been reported. We discuss potential pathomechanisms of this very rare complication and give a review of the literature. PMID- 18392603 TI - [Importance of open and laparoscopic stone surgery]. AB - INTRODUCTION: The increasing spread and technical enhancement of endourological methods has led to displacement of the surgical therapy of renal and ureteral calculi. MATERIALS AND METHODS: Based on a review of current literature, we describe indications, technique, and clinical importance of the open and laparoscopic management of urolithiasis. RESULTS: In Europe and North America, the surgical therapy of urolithiasis only plays a role in cases of very large or hard stones, after failure of shock wave lithotripsy, percutaneous nephrolithotripsy, or ureteroscopic stone removal, and in cases of abnormal renal anatomy, i.e., only in a few percent of all stone therapies. However, in developing countries and emerging markets with different structure and funding of the health care system where the methods of endourology are not readily available, these techniques still have a higher importance. Particularly in Europe, laparoscopic surgery is emerging because calculi can be removed from almost all locations in the kidney and ureter using a transperitoneal or retroperitoneal access. Functional outcomes and complication rates are comparable. The benefits of laparoscopy are less postoperative pain, shorter hospital stay, faster convalescence, and better cosmetic results. CONCLUSIONS: Although procedures for open and laparoscopic removal of renal and ureteral calculi are only performed in rare cases in daily urological practice, they are superior to the endourological techniques in some circumstances. Therefore, they should still be part of the urologist's skills. PMID- 18392604 TI - [Calcium oxalate stone formation. New pathogenetic aspects of an old disease]. AB - Calcium oxalate (CaOx) urolithiasis is a very common disorder. Surprisingly, the pathogenetic mechanisms leading to CaOx stone formation have been largely unknown so far. The long-accepted simple explanation by an exceeding of the solubility product of lithogenic substances in the urine cannot sufficiently describe the complex processes. Deviating from the hypothesis that proclaims that the initial crystal deposition takes place in the lumens of renal tubules, new insights suggest a primary plaque formation in the interstitial space of the renal papilla. Initially, calcium phosphate (CaPh) crystals and organic matrix are deposited along the basement membranes of the thin loops of Henle and extend further in the interstitial space to the urothelium, constituting the so-called Randall's plaques that can be regularly found during endoscopy of CaOx-stone forming patients. These CaPh crystals seem to be the origin for the development of future CaOx stones, which form by the attachment of further matrix molecules and CaOx from the urine to the plaque. The driving forces, the exact pathogenetic mechanisms, and the involved matrix molecules remain largely unknown. Possibly, completely different pathomechanisms lead to the common clinical diagnosis of"CaOx stone former." PMID- 18392606 TI - REML approach for adjusting the Fusarium head blight rating to a phenological date in inoculated selection experiments of wheat. AB - Fusarium head blight is one of the most important wheat diseases causing grain yield and quality losses as well as mycotoxin contamination all over the world. Since Fusarium cannot be reliably controlled with fungicides, breeding has become a favorable tool to decrease the infection severity. In most cases, selection for Fusarium resistance is done by artificial infection in the field. However, there is a risk in preferring late heading genotypes, because heading of wheat is negatively correlated to head blight severity. Because an indirect selection for late maturity is not intended, we considered a statistical approach to avoid this problem. In this paper, we propose a mixed model to analyze extensive Fusarium head blight rating in resistance breeding experiments of wheat. The objective of the analysis was to select for Fusarium resistance, while at the same time ensuring that late heading genotypes, which show less head blight over the shorter vegetation period, are not preferred. Thus, selection was to be done such that genetic variability for heading date was retained. Therefore, the statistical model contained a covariate to adjust for differences in the heading date. The use of covariate adjustment is an easily handled alternative to a bivariate analysis. Covariate adjustment will in practice often work almost equally well as bivariate analysis. Any statistical software with powerful mixed model analysis tools can be used for this type of analysis. We propose an ad hoc method to obtain heritability estimates and a form of LSD (least significance difference) as a measure of accuracy on the basis of the proposed model and under special consideration of the experimental design. The ad hoc LSD was used as a rough measure to judge rankings of genotypic means (BLUPs). Friedman's super smoother was used to compare smoothed rank estimates for adjusted and unadjusted genotypes against increasing smoothed heading dates. Traits were transformed to meet the model assumptions, especially homogeneity of errors and normality, and back-transformation of means and standard errors was conducted by using the delta method. PMID- 18392605 TI - [Is rehabilitation of erectile function following pelvic surgery reasonable? Review of the literature: from sports to PDE5 inhibitors]. AB - Increasingly, urologists are seeing patients with erectile dysfunction after pelvic operations. In most cases, radical prostatectomy is the cause. Even when a nerve-sparing procedure is performed, approximately 50% of the patients suffer from erectile dysfunction. This report discusses the causes and theoretical therapies, including lifestyle changes, strategies for neuroregeneration and the associated prevention of apoptosis of the smooth muscle of the corpus cavernosum and improvement of the corpora cavernosa by increased oxygenation. According to the international literature, many of these agents and lifestyle modifications display promise for treating impotence. Early treatment for patients recovering from pelvic operations seems to be reasonable. It is assumed that the natural recovery of erections may take as long as 18 to 24 months postsurgery or even longer; however, treatment modalities may reduce the time to erectile recovery. PMID- 18392607 TI - Neuropsychological function in children following admission to paediatric intensive care: a pilot investigation. AB - OBJECTIVE: Little is known about neuropsychological status following acute severe paediatric illness. This pilot study explored the effects on memory function of severe acute paediatric illness and associations between memory functioning and psychiatric sequelae. DESIGN AND SETTING: Case control study of children after paediatric intensive care unit (PICU) discharge and healthy volunteers. PATIENTS AND PARTICIPANTS: 16 PICU discharged children comprised of 11 boys and 5 girls (mean age 9.44 +/- 2.85 years) tested a mean of 4.8 +/- 1.4 months following hospital discharge, and 16 age- and sex-matched controls. MEASUREMENTS AND RESULTS: Visual-spatial memory and attention were assessed using the CANTAB battery (visual memory) and verbal memory with the Children Memory Scale; Intelligence Quotient was tested using the Wechsler Abbreviated Scale of Intelligence. Emotional and behavioural function was measured with the Strengths and Difficulties Questionnaire and Impact of Event Scales. Children admitted to PICU displayed statistically poorer performance on tests of spatial memory (spatial working memory) sustained attention (rapid visual information) and verbal memory (word pairs learning and delayed recognition). Septic illness was specifically associated with poorer pattern recognition memory on the CANTAB. There were significant correlations in the PICU group between cognitive functioning and emotional/behaviour scores. CONCLUSIONS: Our results suggest impaired memory and attention in children following acute severe paediatric illness, a specific deficit in children with septic illness and links between memory anomalies and emotional/behavioural problems. The findings and their clinical significance require replication and clarification in a larger sample. PMID- 18392608 TI - Profiling genes in the ICU: are we there yet? PMID- 18392609 TI - Gene profiling in human blood leucocytes during recovery from septic shock. AB - OBJECTIVE: To assess blood leucocytes gene profiling during recovery phase of septic shock; to test the relation between encoding gene expression and protein level. STUDY DESIGN: Gene expression levels were studied at days 0, 1, 7 and 28 (D0, 1, 7 and 28) on a dedicated microarray of 340 genes involved in inflammatory processes. SETTINGS: 16-bed intensive care unit, Lariboisiere University hospital. PATIENTS: Seventeen septic shock patients enrolled when at least one additional organ dysfunction occurred. MEASUREMENTS AND RESULTS: Changes over time were compared with D0 via the ratio Dx/D0. The time-related gene expression study showed significant changes in ten genes. Among them, S100A8 and S100A12 had a reduced expression over time compared with D0, whereas CD74's expression increased. The microarray results were validated by RT-qPCR for four genes. The S100A8 plasma levels decrease along recovery in parallel with the gene expression decrease. The CD74 gene expression evolution significantly correlated with HLA-DR monocyte expression. CONCLUSIONS: These results are the first description of variations in expression of key inflammatory genes in the course of the septic shock recovery period. PMID- 18392610 TI - Two highly thermostable paralogous single-stranded DNA-binding proteins from Thermoanaerobacter tengcongensis. AB - The thermophilic bacterium Thermoanaerobacter tengcongensis has two single stranded DNA-binding (SSB) proteins, designated TteSSB2 and TteSSB3. In a SSB complementation assay in Escherichia coli, only TteSSB3 took over the in vivo function of EcoSSB. We have cloned the ssb genes obtained by PCR and have developed E. coli overexpression systems. The TteSSB2 and TteSSB3 consist of 153 and 150 amino acids with a calculated molecular mass of 17.29 and 16.96 kDa, respectively. They are the smallest known bacterial SSB proteins. The homology between amino acid sequences of these proteins is 40% identity and 53% similarity. They are functional as homotetramers, with each monomer encoding one single-stranded DNA binding domain (OB-fold). In fluorescence titrations with poly(dT), both proteins bind single-stranded DNA with a binding site size of about 40 nt per homotetramer. Thermostability with half-life of about 30 s at 95 degrees C makes TteSSB3 similar to the known SSB of Thermus aquaticus (TaqSSB). The TteSSB2 was fully active even after 6 h incubation at 100 degrees C. Here, we show for the first time paralogous thermostable homotetrameric SSBs, which could be an attractive alternative for known homodimeric thermostable SSB proteins in their applications for molecular biology methods and analytical purposes. PMID- 18392611 TI - Determination of glyphosate and AMPA in surface and waste water using high performance ion chromatography coupled to inductively coupled plasma dynamic reaction cell mass spectrometry (HPIC-ICP-DRC-MS). AB - A novel method employing high-performance cation chromatography in combination with inductively coupled plasma dynamic reaction cell mass spectrometry (ICP-DRC MS) for the simultaneous determination of the herbicide glyphosate (N phosphonomethylglycine) and its main metabolite aminomethyl phosphonic acid (AMPA) is presented. P was measured as (31)P(16)O(+) using oxygen as reaction gas. For monitoring the stringent target value of 0.1 MUg L(-1) for glyphosate, applicable for drinking and surface water within the EU, a two-step enrichment procedure employing Chelex 100 and AG1-X8 resins was applied prior to HPIC-ICP-MS analysis. The presented approach was validated for surface water, revealing concentrations of 0.67 MUg L(-1) glyphosate and 2.8 MUg L(-1) AMPA in selected Austrian river water samples. Moreover, investigations at three waste water treatment plants showed that elimination of the compounds at the present concentration levels was not straightforward. On the contrary, all investigated plant effluents showed significant amounts of both compounds. Concentration levels ranged from 0.5-2 MUg L(-1) and 4-14 MUg L(-1) for glyphosate and AMPA, respectively. PMID- 18392612 TI - Progress in planar electrochromatography. AB - Developments in planar electrochromatography in open (PEC) and closed (PPEC) systems are reviewed. The discussion focuses on progress in chamber construction for planar electrochromatography, separating system performance, equilibration of the PPEC process, separation time and selectivity, and the general advantages, disadvantages and prospects of this separation mode. PMID- 18392613 TI - Circular dichroism thermal lens microscope in the UV wavelength region (UV-CD TLM) for chiral analysis on a microchip. AB - We have developed a circular-dichroism thermal lens microscope for UV wavelengths (UV-CD-TLM), for the first time, to realize sensitive chiral analysis on a microchip. Quasi-continuous-wave phase modulation of a pulsed UV laser was used to generate left-circularly polarized light and right-circularly polarized light and to detect the generated TL signal amplitude and phase with a lock-in amplifier. The amplitude and phase were used to determine the concentration and chirality, respectively, of a sample. The basic principle of UV-CD-TLM for chiral analysis on a microchip was verified by measuring aqueous solutions of optically active camphorsulfonic acids (CSA). Lower limits of detection (LOD) were calculated at S/N = 2 and were 8.7 x 10(-4) mol L(-1) (DeltaA = 5.2 x 10(-6) Abs.) for (+)-CSA and 8.4 x 10(-4) mol L(-1) (DeltaA = 5.0 x 10(-6) Abs.) for (-) CSA. In terms of number of molecules, LODs for UV-CD-TLM were calculated to be 8.7 fmol and 8.4 fmol, respectively. This is at least three orders of magnitude lower than previously obtained. The applicability of UV-CD-TLM for chiral analysis on a microchip was verified. PMID- 18392614 TI - Incubation type Si-based planar ion channel biosensor. AB - A new planar-type ion channel biosensor with the function of cell culture has been fabricated using silicon on an insulator substrate as the sensor chip material. Coating of the sensor chip with fibronectin was essentially important for cell incubation on the chip. Although the seal resistance was quite low (approximately 7 Mohms) compared with the pipette patch-clamp gigaohm seal, the whole-cell channel current of the transient receptor potential vanilloid type 1 (TRPV1) channel expressing HEK293 cells was successfully observed, with a good signal-to-noise ratio, using capsaicin as a ligand molecule. PMID- 18392615 TI - Head, arm and trunk coordination during reaching in children. AB - During postural and locomotor tasks, the orientation of the head with respect to space is maintained in order to serve as an egocentric reference value for maintaining balance. In young children during locomotor tasks, task difficulty determines the coordination of movements between head-trunk segments: the more difficult the task, the more the child limits the head on trunk movement ("en bloc") rather than letting the head move freely in space. For reaching tasks, however, there are no data about the development and maturation of coordination between the head and trunk movements and when the pattern of coordination is considered mature. The goal of this study was to characterize the development of head-trunk coordination during reaching from a sitting position in typically developing children. Forty-four typically-developing (TD) children aged from 2.8 to 11.8 years and six healthy adults participated. Children were divided into five groups (G1-G5) according to their age: 2-3, 4-5, 6-7, 8-9 and 10-11 years old. The task involved reaching towards and grasping a piece of food in the younger group or a wooden block in the older children and adults with the dominant hand, adequate to the grip size of each participant, and returning it to the mouth area to simulate self-feeding. The object was placed in line with the midline of the body at three different distances from the trunk according to the participant's arm length (two within and one beyond arm's length). Rotational movements of the head and trunk in three planes; yaw, roll and pitch, were recorded using three-dimensional tracking systems (Optotrak, Northern Digital, Model 3010 or Ariel Performance Analysis System). The variables analysed were relative head and trunk angle, absolute head and trunk angle, the anchoring index (AI) and initial direction of head and trunk rotation (direction index: DI). Patterns of head-trunk coupling were different along different axes of rotation and across groups. For the AI, a head-stabilized-on-trunk (HST) or "en bloc" pattern was observed with approximately the same frequency as a head-stabilized in-space (HSS) pattern in the youngest children in the yaw plane for reaches within arm's length. In all other planes and for reaches of all distances, a HSS pattern was evident in the youngest children and remained consistent across the groups of children. Compared to the children, adult reaching was characterised by fixed head-trunk coordination (HST) in the roll plane at all reach distances, and greater decoupling in yaw plane motion for the two closest distances. There were no age-related differences in the pitch plane strategy which was mainly HSS. The DI patterns matured by 2-3 or 4-5 years of age, except for reaches to T1 in the pitch plane. In addition, in the roll plane, there was evidence of a two-step maturation that was not complete until adulthood. Maturation of strategies used to stabilize the head and trunk relative to each other and to the reaching arm differ across movement planes for a seated reaching task. Our data suggest that different aspects of head and trunk coordination during reaching movement mature at different rates, like for locomotor tasks previously described, and that the maturation follows a non-chronological and protracted course. These results can serve as a comparative database with which to contrast head and trunk coordination in children with movement disorders. However, in terms of typical development, these data should be considered specific for the task studied and may not reflect general principles of motor development. PMID- 18392616 TI - Enzymatic properties of cellobiose 2-epimerase from Ruminococcus albus and the synthesis of rare oligosaccharides by the enzyme. AB - The gene for cellobiose 2-epimerase (CE) from Ruminococcus albus NE1 was overexpressed in Escherichia coli cells. The recombinant CE was purified to homogeneity by a simple purification procedure with a high yield of 88%, and the molecular mass was 43.1 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis and 44.0 kDa on gel chromatography. It exhibited optimal activity around at 30 degrees C and pH 7.5, and the enzyme activity was inhibited by Al3+, Fe3+, Co2+, Cu2+, Zn2+, Pb2+, Ag+, N-bromosuccinimide, iodoacetate, and 4 chloromercuribenzoate. In addition to cello-oligosaccharides, the enzyme was found to effectively 2-epimerize lactose to yield 4-O-beta-D-galactopyranosyl-D mannose (epilactose), which occurs in cow milk as a rare oligosaccharide. The Km and kcat/Km values toward lactose were 33 mM and 1.6 s(-1) mM(-1), and those toward cellobiose were 13.8 mM and 4.6 s(-1) mM(-1), respectively. N-Acetyl-D glucosamine, uridine 5'-diphosphate-glucose, D-glucose 6-phosphate, maltose, sophorose, laminaribiose, and gentiobiose were inert as substrates for the recombinant CE. We demonstrated that epilactose was resistant to rat intestinal enzymes, utilized by human adult bifidobacteria, and stimulated the tight junction permeability in Caco-2 cells. These results strongly suggest that this rare disaccharide is promising for use as a prebiotic. PMID- 18392617 TI - Perspectives for biotechnological production of biodiesel and impacts. AB - In recent years, biological ways for biodiesel production have drawn an increasing attention and compared to chemical approaches, lipase-mediated alcoholysis for biodiesel production has many advantages. Currently, there are extensive reports about enzyme-mediated alcoholysis for biodiesel production, and based on the application forms of biocatalyst, the related research can be classified into immobilized lipase, whole cell catalyst, and liquid lipase mediated alcoholysis for biodiesel production, respectively. This mini-review is focusing on the study of the aforementioned three forms of biocatalyst for biodiesel production, as well as its impacts and prospects. PMID- 18392618 TI - Progression of intracranial glioma disrupts thymic homeostasis and induces T-cell apoptosis in vivo. AB - The thymus is the site where all T-cell precursors develop, mature, and subsequently leave as mature T-cells. Since the mechanisms that mediate and regulate thymic apoptosis are not fully understood, we utilized a syngenic GL261 murine glioma model to further elucidate the fate of T-cells in tumor bearing C57BL/6 mice. First, we found a dramatic reduction in the size of the thymus accompanied by a decrease in thymic cellularity in response to glioma growth in the brains of affected mice. There was a marked reduction of double positive subset and an increase in the frequency of CD4(+) and CD8(+) single positive T cell subsets. Analysis of double negative thymocytes showed an increase in the accumulation of CD44(+) cells. In contrast, there was a marked loss of CD44 and CD122 expression in CD4(+) and CD8(+) subsets. The growth of intracranial tumors was also associated with decreased levels of HO-1, a mediator of anti-apoptotic function, and increased levels of Notch-1 and its ligand, Jagged-1. To determine whether thymic atrophy could be due to the effect of Notch and its ligand expression by glioma in vivo, we performed a bone marrow transplant experiment. Our results suggest that Notch-1 and its ligand Jagged-1 can induce apoptosis of thymocytes, thereby influencing thymic development, immune system homeostasis, and function of the immune cells in a model of experimental glioma. PMID- 18392619 TI - Vaccine-specific local T cell reactivity in immunotherapy-associated vitiligo in melanoma patients. AB - The occurrence of vitiligo in patients with melanoma is especially reported for patients undergoing immunotherapy. While vitiligo in these patients is thought to be related to an immune response directed against melanoma cells, solid evidence is lacking. Here we report local cytotoxic T cell reactivity in three melanoma patients who developed vitiligo, after experimental immunotherapy using dendritic cell vaccinations. Tetramer analysis showed that vaccine-induced T cells recognizing gp100 and tyrosinase are present at the vitiligo lesions. These T cells secrete IFN-gamma and IL-2 upon peptide specific stimulation as well as upon recognition of the autologous tumor. We show that functional CD8(+) T cells specific for melanoma differentiation antigens used in a melanoma immunotherapy trial, do not only invade the tumor, but also the vitiligo lesions. This directly links vitiligo to the immuno-therapeutic intervention and supports the hypothesis that vitiligo is a marker of immunity against melanoma cells. PMID- 18392620 TI - Follow-up of 11-16 years after modular fixed-bearing TKA. AB - To present representative data of long-term survivorship and clinical outcome for the PFC total knee arthroplasty (PFC-TKA). A consecutive series of 141 TKA was followed for a mean of 13 years (range, 11-16 years). Sixty-five knees were evaluated, 30 of these clinically and radiographically. Twenty-eight knees could only be assessed with the use of a questionnaire. Six patients were living in nursing homes. Fifty-four patients (65 knees) had died. Eleven had undergone a revision. One patient was considered lost to follow-up. With re-operation for any reason as the endpoint, the 10-year survival rate was 92% (n = 91 patients at risk), and the 14-year survival rate was 91% (n = 12). With aseptic loosening of the implant as the endpoint, the 10- and 14-year survival rates were 97%. The mean Knee Society and function scores were 76 and 48 points, respectively. In this multi-surgeon series modular fixed-bearing TKA had good clinical and radiographic results with excellent long-term survivorship. PMID- 18392622 TI - Molecular mechanism of beta-thalassaemia caused by 22-bp duplication. AB - A beta-thalassaemia mutation occurring from insertion of a duplicated 22-bp intron/exon junction of the beta-globin gene has been characterised. The repeated 22-bp insertion causes duplication of a 3' splice site at IVSI/exon 2 junction. Reverse transcription-polymerase chain reaction showed that the proximal 3' splice site present in the duplicated gene is used, leading to a frameshift and a premature chain termination at codon 37. beta-Globin messenger ribonucleic acid (mRNA) transcribed from the mutant gene was not detected, suggesting that the process of nonsense-mediated mRNA decay may be triggered by the premature stop codon. PMID- 18392621 TI - Latissimus dorsi transfer for the treatment of massive tears of the rotator cuff. AB - The objective of this paper was to determine the outcome of the transfer of the latissimus dorsi tendon in patients with massive irreparable rotator cuff tears. Eighteen patients of mean age 54 years (range 37-72 years) with massive irreparable rotator cuff tears were studied. The mean follow-up was 28 months (range 12-58 months). The postoperative Constant score was higher by an average of 21.15 points compared to the preoperative score (P=0.002); 88.8% of patients reported significant pain relief and improved mobility, particularly on external rotation. Postoperative resting antero-posterior radiography in neutral rotation revealed a mean 3.2-mm depression of the humeral head, due to the tenodesis effect of tendon transfer, thus, increasing the deltoid lever arm. These clinical results suggest that latissimus dorsi transfer is a useful surgical technique for treating massive irreparable postero-superior tears of the rotator cuff in young and/or active patients, providing significant pain relief and improved shoulder strength. PMID- 18392623 TI - Current status of epigenetic treatment in myelodysplastic syndromes. AB - Epigenetic deregulation plays an important role in cancer development. The great interest in epigenetics in hematology and oncology results from the fact that epigenetic, in contrast to genetic, alterations are, in principle, amenable to pharmacological reversal. Epigenetically active drugs currently within clinical trials include histone deacetylase inhibitors (HDACi) and DNA methyltransferase (DNMT) inhibitors. The first treatment approved by the Food and Drug Administration for the treatment of myelodysplastic syndromes (MDS) was the DNMT inhibitor 5-azacytidine. Currently, two out of three drugs FDA approved for MDS therapy, 5-azacytidine and 5-aza-2'-deoxycytidine, are epigenetically active drugs. Recent clinical trials investigate new dosing schedules, routes of administration, and combination regimens. Several structurally distinct HDACi have been developed. Available data is mostly restricted to phase I trials. The largest experience in MDS and acute myeloid leukemia exists with the anticonvulsant valproic acid. This review summarizes the existing clinical experience on HDACi and DNMT inhibitors. PMID- 18392624 TI - Characterization and expression of plasma and tonoplast membrane aquaporins in elongating cotton fibers. AB - Cotton fiber (Gossypium hirsutum L. and G. barbadense L.) is a good model for studies of plant cell elongation and cell wall biogenesis. Aquaporins are ancient membrane channel proteins that facilitate the permeation of water across biological membranes. We studied GhPIP1-2, encoding plasma membrane intrinsic protein, and GhgammaTIP1, encoding tonoplast intrinsic protein, during cotton fiber development. The full-length cDNAs of GhPIP1-2 and GhgammaTIP1 were obtained through 5' RACE. The deduced amino acid sequences of GhPIP1-2 and GhgammaTIP1 share high sequence identity with aquaporins from diverse plant species. Phylogenetic analysis of GhPIP1-2 and GhgammaTIP1 with other plant aquaporins showed that GhPIP1-2 belongs to the PIP1 group of the PIP subfamily and GhgammaTIP1 belongs to the gammaTIP group of the TIP subfamily. GhPIP1-2 and GhgammaTIP1 contain three and two introns, respectively. Genomic Southern blot analysis indicated that GhPIP1-2 and GhgammaTIP1 have several copies and multiple homologous genes in allotetraploid cotton. Northern blot analysis with gene specific probes and real-time PCR demonstrated that GhPIP1-2 and GhgammaTIP1 are predominantly expressed during cotton fiber elongation, with the highest expression levels at 5 days post-anthesis. Moreover, expression patterns of the two genes in G. hirsutum and G. barbadense are similar, whereas the expression levels in G. barbadense are much lower than that in G. hirsutum. The high and preferential expression of GhPIP1-2 and GhgammaTIP1 during fiber cell elongation suggests that they may play important roles in supporting the rapid influx of water into vacuoles during cotton fiber cell expansion. PMID- 18392625 TI - Overexpression of the maize Teosinte Branched1 gene in wheat suppresses tiller development. AB - The number of viable shoots influences the overall architecture and productivity of wheat (Triticum aestivum L.). The development of lateral branches, or tillers, largely determines the resultant canopy. Tillers develop from the outgrowth of axillary buds, which form in leaf axils at the crown of the plant. Tiller number can be reduced if axillary buds are not formed or if the outgrowth of these buds is restricted. The teosinte branched1 (tb1) gene in maize, and homologs in rice and Arabidopsis, genetically regulate vegetative branching. In maize, increased expression of the tb1 gene restricts the outgrowth of axillary buds into lateral branches. In this study, the maize tb1 gene was introduced through transformation into the wheat cultivar "Bobwhite" to determine the effect of tb1 overexpression on wheat shoot architecture. Examination of multiple generations of plants reveals that tb1 overexpression in wheat results in reduced tiller and spike number. In addition, the number of spikelets on the spike and leaf number were significantly greater in tb1-expressing plants, and the height of these plants was also reduced. These data reveal that the function of the tb1 gene and genetic regulation of lateral branching via the tb1 mode of action is conserved between wheat, rice, maize and Arabidopsis. Thus, the tb1 gene can be used to alter plant architecture in agriculturally important crops like wheat. PMID- 18392626 TI - Dual targeting of organellar seryl-tRNA synthetase to maize mitochondria and chloroplasts. AB - Aminoacyl-tRNA synthetases (AARSs) play a critical role in translation and are thus required in three plant protein-synthesizing compartments: cytosol, mitochondria and plastids. A systematic study had previously shown extensive sharing of organellar AARSs from Arabidopsis thaliana, mostly between mitochondria and chloroplasts. However, distribution of AARSs from monocot species, such as maize, has never been experimentally investigated. Here we demonstrate dual targeting of maize seryl-tRNA synthetase, SerZMo, into both mitochondria and chloroplasts using combination of complementary methods, including in vitro import assay, transient expression analysis of green fluorescent protein (GFP) fusions and immunodetection. We also show that SerZMo dual localization is established by the virtue of an ambiguous targeting peptide. Full-length SerZMo protein fused to GFP is targeted to chloroplast stromules, indicating that SerZMo protein performs its function in plastid stroma. The deletion mutant lacking N-terminal region of the ambiguous SerZMo targeting peptide was neither targeted into mitochondria nor chloroplasts, indicating the importance of this region in both mitochondrial and chloroplastic import. PMID- 18392627 TI - Ki67 staining index and neuroendocrine differentiation aggravate adverse prognostic parameters in prostate cancer and are characterized by negligible inter-observer variability. AB - INTRODUCTION: This study aims to clarify whether neuroendocrine differentiation (NED) and/or proliferation activity assessed by means of Ki67 staining index (Ki67SI) might aggravate other established adverse prognostic parameters commonly used for predicting outcome in surgically treated prostate cancer, and to assess inter-observer variability in assigning NED and Ki67 SI. MATERIAL AND METHODS: A total of 528 patients surgically treated due to prostate cancer were evaluated in this study. Relevant data were retrospectively obtained by chart review. Immunostaining with antibodies directed against Chromogranin A and Ki67 was performed on archived surgical material, and was evaluated by two independent histopathologists blinded to the specimens. Surveying a median postsurgical follow-up of 46.4 months, postsurgical serum PSA-levels were regularly documented for identifying biochemical progress. Multivariate analysis was performed by means of the Cox regression hazards regression method to evaluate possible aggravations of established adverse prognostic parameters (nodal status, tumour stage, pretherapeutic PSA-level, and Gleason score) by NED and/or Ki67SI. Ki67 SI and NED were shown to significantly aggravate these established adverse prognostic parameters, and were found to be characterized by negligible inter observer variability. CONCLUSION: Ki67 SI and NED should be advocated to be rendered by the histopathologist because both parameters can be immunohistochemically determined without much additional expense in time and cost involved. This concept is rewarded by an additional gain of prognostic accuracy in evaluating individual risk profile after surgery. PMID- 18392628 TI - [Mucous membrane pemphigoid with ocular involvement. Part II: therapy]. AB - Treatment of mucous membrane pemphigoid (MMP) aims at reduction of conjunctival inflammation by means of systemic immunosuppression. In addition, cicatricial progression and management of the resulting ocular surface disease requires topical conservative or surgical measures. The former includes systemic immunosuppression with steroids and other immunosuppressive agents: dapsone in mild to moderate disease and cyclophosphamide in severe cases have been established in two randomized trials. Other agents such as methotrexate, azathioprine, mycophenolate mofetil or monoclonal antibodies including daclizumab or rituximab were found to be effective in uncontrolled small studies. Surgery is primarily focused on eyelid problems such as entropium and trichiasis. Ocular surface disease and secondary complications, e.g. cataract formation and glaucoma, may need surgical treatment. Any surgery is associated with the risk of a relapse of inflammation and should be postponed until inflammation is controlled by systemic therapy. Management of MMP patients requires close collaboration of a specialized ophthalmologist with specialists from dermatology and internal medicine. PMID- 18392629 TI - Congenital cystic lung disease: contemporary antenatal and postnatal management. AB - Congenital cystic lung disease comprises a broad spectrum of rare but clinically significant developmental abnormalities, including congenital pulmonary adenomatoid malformations, bronchopulmonary sequestrations, bronchogenic cysts, and congenital lobar emphysema that result from perturbations in lung and airway embryogenesis. As congenital lung lesions are now more commonly recognized antenatally, mothers require accurate prenatal counseling and appropriate perinatal management. In light of long-term complications of infection and malignancy, there is growing consensus that infants with asymptomatic lesions should undergo elective excision of congenital pulmonary adenomatoid malformation (CPAM) or bronchopulmonary sequestration (BPS). This review will focus on advancements and current practice in the diagnosis and management of CPAM and BPS, identifying aspects of the literature that are confusing or controversial. Although our knowledge and pre- and postnatal management of lung lesions will continue to evolve and improve, there is a compelling need for a unified clinical and pathological classification system that creates a common platform for discussion, clinical management, and research. PMID- 18392630 TI - Adriamycin produces a reproducible teratogenic model of gastrointestinal atresia in the mouse. AB - Gastrointestinal atresia is a major cause of bowel obstruction in the newborn. Experimental models and clinical observations have demonstrated the heterogeneous nature of its pathogenesis. A proportion is due to late intra-uterine vascular insults and some are genetic in nature. Epidemiological studies have found gastrointestinal atresia to occur with other birth defects, in particular VACTERL anomalies, suggesting that a subset of cases may result from an early disturbance to intestinal morphogenesis. Adriamycin is teratogenic in rats, producing gastrointestinal atresia and VACTERL anomalies. The mouse is the foremost mammal studied by developmental biologists, offering an expanding wealth of knowledge and scientific research techniques. The aim of this study was to create an Adriamycin mouse model for investigating the development of gastrointestinal atresia. CBA/Ca mice were accurately time-mated (n = 30). Four different doses of Adriamycin (0-saline control, 4, 5 and 6 mg/kg) at three different timings of injections were compared (12 groups). Dams received two intraperitoneal injections, 24 h apart, commencing on day 7, 7.5 or 8. Foetuses were harvested on day 18. Gastrointestinal atresia and VACTERL anomalies were examined using a dissecting microscope. Adriamycin produced type IIIa gastrointestinal atresia in six treatment groups. The effect of Adriamycin depended on the timing and dose of the injections. VACTERL anomalies were only found in four treatment groups, proposing overlapping critical embryological windows for these malformations. Gastrointestinal atresia can be induced by the teratogen Adriamycin, occurring with and without VACTERL anomalies. This produces a reproducible mouse model in which the molecular pathogenesis of gastrointestinal atresia may be studied. PMID- 18392631 TI - Early complications of the Nuss procedure for pectus excavatum: a prospective study. AB - The Nuss procedure is a minimally invasive method for the correction of pectus excavatum, with several centers reporting its successful application. Complications related to the Nuss procedure are not uncommon and life-threatening complications have been reported. This study focuses on the incidence and management of complications in a series of 167 children and adults with funnel chest corrected by Nuss procedure. Guidelines and strategies to avoid the most common and typical complications are proposed. All patients with funnel chest, operated between April 2000 and 2006 were evaluated prospectively. Our surgical approach involved the submuscular insertion of the pectus bar under right-sided thoracoscopic control. The bar was secured in most cases with one stabilizer on the right side on the underlying rib to prevent bar displacement. Postoperative pain was primarily managed by epidural catheters. All data in the patient report forms was prospectively entered in a database. All complications were documented and classified into major or minor complication. A major complication was noted, if an organ injury occurred or if a significant surgical intervention became necessary. A minor complication was documented, if either an endoscopy or an evacuation of fluid or gas from the thorax by puncture were necessary. One hundred and sixty seven patients (136 males and 31 females) with a mean age of 16.3 (range 5-40 years) were included in this study. Major complications occurred in seven patients (4.2%) and consisted of one intraoperative heart perforation, one piercing of the liver with the trocar, bar infections (n = 2) and significant bar displacement (n = 3). Minor complications were seen in 122 patients (73.1%) and consisted of breakage of wires used to secure the lateral stabilizer plate (n = 48), pleural effusions (n = 28), intraoperative rupture of the intercostal muscle (n = 15), pericardial tears without clinical significance (n = 7) and lung atelectasia (n = 4). Major complications related to the Nuss procedure were rare but preventable and could mainly be attributed to the learning curve. Most minor complications can be avoided by changing the technique, e.g. fixation of the bar and the stabilizer onto the underlying rib, use of PDS cords instead of metal wires to fix the bar and the stabilizer, entrance into and exit of the thorax medial to the rim of the pectus excavatum, etc. Some complications are related to the technique, such as minor pleural effusion or remaining gas in the thorax. Clear guidelines in regard to the technique are presented to prevent the majority of complications and thereby shorten the learning curve. PMID- 18392632 TI - The impact of treatment center on the outcome of patients with laryngeal cancer treated with surgery and radiotherapy. AB - For laryngeal cancer, surgical excision of the primary tumor should be undertaken with the aim of achieving tumor-free margins. Adequate pathological assessment of the specimen and the competency of the treatment center play a crucial role in achieving cure. The present study aimed to analyze the significance of place of surgery on the outcome of patients with laryngeal cancer who underwent surgical operation in other centers and were subsequently referred to Dokuz Eylul University Head and Neck Tumour Group (DEHNTG) for postoperative irradiation. Patients were divided into three groups according to their place of surgery. The first group (Group I) consisted of patients who had their surgical operation at DEUH. Patients in the second group (Group II) were referred from centers with oncological surgical experience. The third group (Group III) consisted of patients referred from hospitals with no surgical teams experienced in head and neck cancer treatment. The clinical and pathological features of patients in these three groups were analyzed to assess the impact of place of surgery on clinical outcome as well as the prognostic factors for survival. The study population consisted of 253 patients who were treated between 1991 and 2006 with locally advanced laryngeal cancer according to the protocol of DEHNTG. The median follow-up was 48 (3-181) months. The 5 years overall, loco-regional disease-free and distant disease-free survivals were 66, 88 and 91%, respectively. When patients' clinical and histopathological features were analyzed for the impact of place of surgery, surgical margin positivity rates were found to be higher in Group III (P = 0.032), although the other two groups had more advanced clinical and pathological N stage disease (P = 0.012, P = 0.001). In multivariate analysis, older age (P < 0.0001), presence of perinodal invasion (P = 0.012), time interval between surgery and radiotherapy longer than 6 weeks (P = 0.003) and tumor grade (P = 0.049) were the most significant factors. For loco-regional failure-free survival, advanced clinical stage (P = 0.002), place of surgery (P = 0.031) and presence of clinical subglottic invasion (P = 0.029) were shown to be important prognostic factors. For distant metastasis-free survival, only pathological (+) lymph node status (P = 0.046) was a significant factor in multivariate analysis. The significance of place of surgery as well as other well known prognostic factors underlines the importance of an experienced multidisciplinary treatment team if best results are to be obtained for the patient. PMID- 18392633 TI - Content comparison of quality of life questionnaires used in head and neck cancer based on the international classification of functioning, disability and health: a systematic review. AB - The objective of this study is to provide a content comparison of frequently used questionnaires that assess health-related quality of life (hrQOL) in head and neck cancer (HNC) survivors. This systematic content comparison describes which specific areas of hr-QOL research are covered by each questionnaire. Thereby, it shall assist the clinician in the decision process of instrument selection depending on the content of the study question. As a reference, we chose the international classification of functioning, disability and health (ICF), which was adopted by the WHO in 2001. A systematic literature review identified current hrQOL questionnaires relevant for HNC. The concepts of functioning contained in each questionnaire were translated (linked") to the ICF according to standardized guidelines. Nine questionnaires were selected for further analyses: EORTC-QLQ (C30 + HN35), FACT (G + HN), UW_QOL, QOL-RTI, HN-QOL, PSS-HN, VHI, LORQ, XQ. Within the selected questionnaires, there are 474 concepts, matching 74 second level ICF categories. The results are presented in tables, showing for each of the validated questionnaires, which of these 74 categories of functioning are addressed. In terms of diversification of content among the questionnaires, there are just eight categories that are used rather frequently and apply to at least five (out of nine) of the questionnaires: e110 Products for personal consumption (i.e., food, drugs), b510 ingestion function, b152 emotional function, b280 sensation of pain, b310 voice, d550 eating, b130 energy and drive function and d850 employment. This ICF-based content comparison provides detailed information on the content that is covered in each questionnaire and thereby assists questionnaire selection. The results question the assumption that HNC-specific questionnaires generally cover the same content. Depending on the study question, the population to be studied and the intervention, there is no unique ideal questionnaire. Compared with other types of qualitative review, the most important advantage of content comparison based on the ICF is the use of an external and independent reference. PMID- 18392635 TI - Comment on "Neutron-induced 63Ni in copper samples from Hiroshima and Nagasaki: a comprehensive presentation of results obtained at the Munich Maier-Leibnitz Laboratory" by W. Ruhm, K. L. Carroll, S. D. Egbert, T. Faestermann, K. Knie, G. Korschinek, R. E. Martinelli, A. A. Marchetti, G. Rugel, T. Straume, A. Wallner, C. Wallner, S. Fujita, H. Hasai, M. Hoshi, K. Shizuma (2007) Radiat Environ Biophys 46:327-338. PMID- 18392636 TI - Ultrastructural localization and chemical binding of silver ions in human organotypic skin cultures. AB - Organotypic cultures of human breast skin incubated with silver bandage or treated with silver sulfadiazine accumulated silver in epithelial cells and in macrophages, fibroblasts and collagen fibrils and fibres of underlying connective tissue. Ultrastructurally, the accumulated silver was found in lysosome-like vesicles of the different cells and evenly spread along collagen structures. Apoptotic nuclei were present but few. Autometallographic amplification of 2D PAGE gels revealed that glutathione S-transferase and glutathion detoxify silver ions in the epidermal cell by binding them in silver-sulphur nanocrystals. Thus, the cytotoxic effect of silver ions seems to be muted by silver ions by being: (1) taken up by undamaged cells, neutralised by glutathione (GSH) and accumulated in lysosomal vesicles, (2) bound extracellularly to SH-groups of the collagen fibres. PMID- 18392637 TI - The role of COX-2 in rectal cancer treated with preoperative radiotherapy. AB - Radiotherapy is one of the principal modalities of rectal cancer treatment, and the ability to predict radio resistance could potentially improve survival through a targeted treatment approach. Cyclooxygenase-2 (COX-2) may protect against damage by irradiation that would justify the use of COX-2 inhibitors. The purpose of this study was to investigate the potential role of COX-2 in tumor response and outcome of patients with rectal cancer treated preoperatively with radiotherapy. Using immunohistochemistry, we examined COX-2 expression in 88 surgical specimens of rectal cancer treated preoperatively and in 26 pretherapeutic biopsies. We tested whether COX-2 expression was correlated with clinico-pathologic parameters and with survival and local recurrence. COX-2 was expressed in 50% of the pretherapeutic tumor biopsies and in 88.6% of post irradiated surgical samples. COX-2 expression was correlated only with enhanced tumor inflammation (p = 0.03) and with tumor volume exceeding 30 cc (p = 0.05). COX-2 was not significantly correlated with patient survival, but none of the patients with COX-2 negative tumors did recur locally, whereas 80% of patients with local recurrences have COX-2 positive tumors. We conclude that COX-2 expression is overexpressed in the majority of rectal cancers treated with radiotherapy and likely plays a role in local relapse. PMID- 18392638 TI - Vertical transmission of HIV in Belgium: a 1986-2002 retrospective analysis. AB - Prophylactic interventions have lead to the reduction of the mother-to-child transmission (MTCT) of human immunodeficiency virus type 1 (HIV-1) to less than 2% in industrialized countries. The aim of this study was to evaluate the changes over time in vertical transmission according to the standard care of prophylaxis in the practice of a single large reference center and to identify the risk factors for failure. The rate of MTCT decreased progressively from 10% in 1986 1993 to 4.7% in 1999-2002, reflecting the progressive implementation of newly available means of prevention. During the last period evaluated (1999-2002), where highly active antiretroviral therapy (HAART) prophylaxis was the standard of care, 17% of women had a viral load between 400 and 20,000 copies/ml around delivery and 5% had a viral load above 20,000 copies/ml. High viral load and low CD4 lymphocyte count were strongly associated with vertical transmission. The rate of MTCT in women who received HAART for more than one month during pregnancy was 1.7%, compared to 13.3% in women treated with HAART for less than one month. The risk of vertical transmission in the absence of therapy was four times higher than before the era of antiretroviral therapy (ART; p=0.05). In conclusion, since the prevention of MTCT of HIV with HAART is the standard of care, a short duration or absence of ART during pregnancy linked to late or absent prenatal care is associated with a high risk of transmission. The early detection of HIV-1 infection in pregnant women, and close follow up and support during pregnancy are crucial to the success of the prevention of transmission. PMID- 18392639 TI - QTc interval prolongation in children with Turner syndrome: the results of exercise testing and 24-h ECG. AB - BACKGROUND: Turner syndrome (TS) is the most common sex chromosome abnormality in females. Recently, a prolongation of the rate-corrected QT (QTc) interval in the electrocardiogram (ECG) of TS patients has been reported. A prolonged QTc interval has been correlated to an increased risk for sudden cardiac death, and medical treatment is warranted in patients with congenital long QT syndrome (LQTS). Additionally, several drugs of common use are contraindicated in LQTS because of their effects on myocardial repolarization. The importance of the QTc prolongation in TS patients is not known at present. MATERIALS AND METHODS: Eighteen TS patients with a prolonged QTc interval (group 1) and 11 TS patients with a normal QTc interval (group 2) (mean age 12.6+/-3.1 vs. 11.8+/-2.1 years, respectively) were tested. The QTc interval was calculated during exercise testing and during 24-h ECG recordings. RESULTS: None of the patients experienced adverse cardiac events during the tests. The mean QTc interval decreased from 0.467 to 0.432 s in group 1 and from 0.432 to 0.412 s in group 2. During the 24-h ECG, the maximum QTc interval was significantly prolonged in group 1 (0.51 vs. 0.465 s, p<0.05, respectively). We conclude that exercise testing and 24-h ECG recording provide valuable information about the cardiac risk in the single TS patient with a prolonged QTc interval. This helps in counseling these girls, as clear therapeutic guidelines are currently lacking. PMID- 18392640 TI - Presumptive varicella vaccination is warranted in Greek adolescents lacking a history of disease or household exposure. AB - Current practice favors serotesting adolescents with a negative history of chickenpox rather than offering presumptive vaccination. Recent epidemiologic data from Greece indicate that a high proportion of adolescents (21.5%) are susceptible to chickenpox. We assessed the reliability of negative varicella history in relation to type of exposure in 311 children and 283 adolescents. In children with social or unknown exposure to varicella, a negative history had a high negative predictive value (NPV = 73.5), supporting the clinical practice of presumptive vaccination. Conversely, children with a negative history and household exposure had a low NPV (13.8), suggesting that pre-vaccination serologic testing is warranted. In conclusion, based on our local epidemiologic data, presumptive varicella vaccination should be offered to all adolescents with the exception of the subgroup of adolescents with household exposure. PMID- 18392641 TI - 15-Lipoxygenase gene variants are associated with carotid plaque but not carotid intima-media thickness. AB - The major underlying cause of CHD is atherosclerosis, and oxidised LDL is known to play an important role in its development. We examined the role of three single nucleotide polymorphisms (SNPs) in the 15-lipoxygenase gene (ALOX15), in atherosclerosis. We genotyped three SNPs in the ALOX15 promoter in two Western Australian samples-1,111 community-based individuals and 556 with CHD. SNPs and haplotypes were tested for an association with carotid plaque, intima-media thickness and risk of CHD. The -611GG genotype was associated with increased likelihood of carotid plaque in CHD patients (OR = 4.01, 95%CI = 1.39-11.53, P = 0.005) and the C alleles of the G-220C and G-189C SNPs were associated with decreased likelihood of plaque among cases (OR = 0.66, 95%CI = 0.43-0.99, P = 0.05 and OR = 0.51, 95%CI = 0.34-0.78, P = 0.002 respectively). The GGG haplotype was associated with increased risk of carotid plaque in CHD patients (OR = 5.77, 95%CI = 1.82-18.29, P = 0.0007) and in community-based individuals under 53 years (OR = 4.15, 95%CI = 1.23-14.08, P = 0.02). No association was observed between ALOX15 SNPs or haplotypes and intima-media thickness. This study is novel as it is the first to examine the association between 15-lipoxygenase polymorphisms and atherosclerotic indicators. These findings suggest a possible role of ALOX15 polymorphisms in focal plaque formation. PMID- 18392643 TI - NPHS2 screening with SURVEYOR in Hellenic children with steroid-resistant nephrotic syndrome. PMID- 18392642 TI - Temporal dynamics of the carbon isotope composition in a Pinus sylvestris stand: from newly assimilated organic carbon to respired carbon dioxide. AB - The (13)C isotopic signature (C stable isotope ratio; delta(13)C) of CO(2) respired from forest ecosystems and their particular compartments are known to be influenced by temporal changes in environmental conditions affecting C isotope fractionation during photosynthesis. Whereas most studies have assessed temporal variation in delta(13)C of ecosystem-respired CO(2) on a day-to-day scale, not much information is available on its diel dynamics. We investigated environmental and physiological controls over potential temporal changes in delta(13)C of respired CO(2) by following the short-term dynamics of the (13)C signature from newly assimilated organic matter pools in the needles, via phloem-transported organic matter in twigs and trunks, to trunk-, soil- and ecosystem-respired CO(2). We found a strong 24-h periodicity in delta(13)C of organic matter in leaf and twig phloem sap, which was strongly dampened as carbohydrates were transported down the trunk. Periodicity reappeared in the delta(13)C of trunk respired CO(2), which seemed to originate from apparent respiratory fractionation rather than from changes in delta(13)C of the organic substrate. The diel patterns of delta(13)C in soil-respired CO(2) are partly explained by soil temperature and moisture and are probably due to changes in the relative contribution of heterotrophic and autotrophic CO(2) fluxes to total soil efflux in response to environmental conditions. Our study shows that direct relations between delta(13)C of recent assimilates and respired CO(2) may not be present on a diel time scale, and other factors lead to short-term variations in delta(13)C of ecosystem-emitted CO(2). On the one hand, these variations complicate ecosystem CO(2) flux partitioning, but on the other hand they provide new insights into metabolic processes underlying respiratory CO(2) emission. PMID- 18392644 TI - Glomerulocystic kidney disease in hypomelanosis of Ito. AB - Hypomelanosis of Ito (incontinentia pigmenti type I) was first described as a disorder characterized by unusual unilateral or bilateral cutaneous macular hypopigmented whorls, streaks and patches. Subsequently, neurologic, skeletal and ocular involvement were described. Kidney involvement has been reported only exceptionally. Here, we describe the case of a male infant with hypomelanosis of Ito with a prenatal diagnosis of bilateral enlargement of the kidneys, decreased corticomedullary differentiation and cysts located in the cortical and subcapsular regions. These findings were confirmed postnatally. The skin examination showed hypopigmented linear and round diffuse lesions located on the right leg and the arms. Ophthalmological examination showed anterior capsular and posterior subcapsular cataract of the left eye. Renal biopsy was characteristic of glomerulocystic kidney disease, whereas the skin biopsy confirmed the clinical diagnosis of hypomelanosis of Ito. Four other cases of kidney disease in hypomelanosis of Ito have been reported, including two cases characterized by cystic renal changes, indicating that gene abnormalities that cause hypomelanosis of Ito may also impair normal renal development, causing renal cystic changes. PMID- 18392646 TI - [Charcot arthropathy--a challenge]. PMID- 18392645 TI - AMF-induced biocontrol against plant parasitic nematodes in Musa sp.: a systemic effect. AB - Although mycorrhizal colonization provides a bioprotectional effect against a broad range of soil-borne pathogens, including plant parasitic nematodes, the commercial use of arbuscular mycorrhizal fungi (AMF) as biocontrol agents is still in its infancy. One of the main reasons is the poor understanding of the modes of action. Most AMF mode of action studies focused on AMF-bacterial/fungal pathogens. Only few studies so far examined AMF-plant parasitic nematode interactions. Therefore, the aim of the study was to determine whether the AMF Glomus intraradices was able to incite systemic resistance in banana plants towards Radopholus similis and Pratylenchus coffeae, two plant parasitic nematodes using a split-root compartmental set-up. The AMF reduced both nematode species by more than 50%, even when the AMF and the plant parasitic nematodes were spatially separated. The results obtained demonstrate for the first time that AMF have the ability to induce systemic resistance against plant parasitic nematodes in a root system. PMID- 18392650 TI - Ultra high field MR: useful instruments or toys for the boys. PMID- 18392651 TI - Focused approach to parathyroidectomy. AB - Minimally invasive parathyroidectomy is performed after preoperative parathyroid localization-usually with high-quality sestamibi scans and/or ultrasonography often under cervical block anesthesia during which a limited exploration is performed. The rapid intraoperative parathyroid hormone assay is then employed to confirm an adequate resection and cure of primary hyperparathyroidism. This article discusses imaging, anesthesia, results, and the surgical management of patients undergoing minimally invasive parathyroidectomy. PMID- 18392652 TI - Optimal depth of NIM EMG endotracheal tube for intraoperative neuromonitoring of the recurrent laryngeal nerve during thyroidectomy. AB - BACKGROUND: Malpositioning of the endotracheal surface electrodes can result in dysfunction of intraoperative neuromonitoring (IONM) and increase the risk of recurrent laryngeal nerve injury. The purpose of this study was to investigate the optimal depth of the nerve integrity monitor (NIM) EMG endotracheal tube. METHODS: We enrolled 105 adult patients undergoing elective thyroidectomy. Each Medtronic Xomed NIM EMG endotracheal tube was placed with the middle of the exposed electrodes well in contact with the true vocal cords under direct laryngoscopy. Function of IONM was documented and the insertion depth was measured and analyzed. RESULTS: Ninety-nine (94.3%) patients had successful IONM with the initial endotracheal tube position. Six (5.7%) patients needed further tube depth adjustment under fiberoptic bronchoscopy. All patients were finally had successful IONM. The optimal mean depth was 20.6 +/- 0.97 cm in men and 19.6 +/- 1.0 cm in women (p < 0.01). There was the trend that taller subjects had a deeper tube depth (p < 0.05). CONCLUSION: We concluded that the mean depth of the NIM EMG tube would be a useful reference value for detecting the malposition of electrodes and adjusting the depth of tube during the operation. PMID- 18392653 TI - Genetic background determines metabolic phenotypes in the mouse. AB - To evaluate the contribution of genetic background to phenotypic variation, we compared a large range of biochemical and metabolic parameters at different ages of four inbred mice strains, C57BL/6J, 129SvPas, C3HeB/FeJ, and Balb/cByJ. Our results demonstrate that important metabolic, hematologic, and biochemical differences exist between these different inbred strains. Most of these differences are gender independent and are maintained or accentuated throughout life. It is therefore imperative that the genetic background is carefully defined in phenotypic studies. Our results also argue that certain backgrounds are more suited to study a given physiologic phenomenon, as distinct mouse strains have a different propensity to develop particular biochemical, hematologic, and metabolic abnormalities. These genetic differences can furthermore be exploited to identify new genes/proteins that contribute to phenotypic abnormalities. The choice of the genetic background in which to generate and analyze genetically engineered mutant mice is important as it is, together with environmental factors, one of the most important contributors to the variability of phenotypic results. PMID- 18392654 TI - Characterization of mouse Dactylaplasia mutations: a model for human ectrodactyly SHFM3. AB - SHFM3 is a limb malformation characterized by the absence of central digits. It has been shown that this condition is associated with tandem duplications of about 500 kb at 10q24. The Dactylaplasia mice display equivalent limb defects and the two corresponding alleles (Dac1j and Dac2j) map in the region syntenic with the duplications in SHFM3. Dac1j was shown to be associated with an insertion of an unspecified ETn-like mouse endogenous transposon upstream of the Fbxw4 gene. Dac2j was also thought to be an insertion or a small inversion in intron 5 of Fbxw4, but the breakpoints and the exact molecular lesion have not yet been characterized. Here we report precise mapping and characterization of these alleles. We failed to identify any copy number differences within the SHFM3 orthologous genomic locus between Dac mutant and wild-type littermates, showing that the Dactylaplasia alleles are not associated with duplications of the region, in contrast with the described human SHFM3 cases. We further show that both Dac1j and Dac2j are caused by insertions of MusD retroelements that share 98% sequence identity. The differences between the nature of the human and mouse genomic abnormalities argue against models proposed so far that either envisioned SHFM3 as a local trisomy or Dac as a mutant allele of Fbxw4. Instead, both genetic conditions might lead to complex alterations of gene regulation mechanisms that would impair limb morphogenesis. Interestingly, the Dac2j mutation occurs within a highly conserved element that may represent a regulatory sequence for a neighboring gene. PMID- 18392655 TI - Susceptibility of four inbred mouse strains to a low-pathogenic isolate of Yersinia enterocolitica. AB - EUMORPHIA (European Union Mouse Research for Public Health and Industrial Application) is a research program involved in developing new approaches in phenotyping, mutagenesis, and informatics to improve characterization of mouse models for understanding human physiology and disease. Secondary screen experiments include the development of assays to identify mice with altered susceptibility or resistance to infections. In this context we developed a new model and established a standard operating procedure for the experimental infection of mice with Yersinia (Y.) enterocolitica. In contrast with previous studies that dealt with high-pathogenic Y. enterocolitica, we used the low pathogenic Y. enterocolitica strain E40 to analyze differences in the immune response of four strains of inbred mice (BALB/c, C3H/HeN, 129P2, C57BL/6) after oral infection. The determination of colony-forming units in Peyer's patches and histologic analysis supported the observations that BALB/c are less able to ameliorate the infection within 21 days. The immune defense of C57BL/6 mice against Yersinia was the most effective resulting in a nearly complete elimination of bacteria after 21 days. C3H/HeN and 129P2 were intermediate. Analysis of serum immunoglobulins (Ig) by Luminex showed a significant increase of IgG2b levels 21 days after infection in all four inbred strains. The other immunoglobulins remained nearly constant. Our infection model discriminates between the efficiency of an infection at an early time point (3 days) and immunity at a later time point (21 days). It is furthermore an appropriate model to characterize genetic differences in resistance and immunity of inbred and mutant mouse lines. PMID- 18392657 TI - Development of a Google-based search engine for data mining radiology reports. AB - The aim of this study is to develop a secure, Google-based data-mining tool for radiology reports using free and open source technologies and to explore its use within an academic radiology department. A Health Insurance Portability and Accountability Act (HIPAA)-compliant data repository, search engine and user interface were created to facilitate treatment, operations, and reviews preparatory to research. The Institutional Review Board waived review of the project, and informed consent was not required. Comprising 7.9 GB of disk space, 2.9 million text reports were downloaded from our radiology information system to a fileserver. Extensible markup language (XML) representations of the reports were indexed using Google Desktop Enterprise search engine software. A hypertext markup language (HTML) form allowed users to submit queries to Google Desktop, and Google's XML response was interpreted by a practical extraction and report language (PERL) script, presenting ranked results in a web browser window. The query, reason for search, results, and documents visited were logged to maintain HIPAA compliance. Indexing averaged approximately 25,000 reports per hour. Keyword search of a common term like "pneumothorax" yielded the first ten most relevant results of 705,550 total results in 1.36 s. Keyword search of a rare term like "hemangioendothelioma" yielded the first ten most relevant results of 167 total results in 0.23 s; retrieval of all 167 results took 0.26 s. Data mining tools for radiology reports will improve the productivity of academic radiologists in clinical, educational, research, and administrative tasks. By leveraging existing knowledge of Google's interface, radiologists can quickly perform useful searches. PMID- 18392658 TI - Relationships between sex and stress hormone levels in feces and marking behavior in a wild population of Iberian wolves (Canis lupus signatus). AB - Feces deposited by the breeding alpha pair on exposed substrates and/or zones may act as visual and olfactory marks associated with social dominance in wolves. The aim of this study was to determine if there was a correlation between marking behavior, sex hormone levels, and physiological stress in a wild population of Iberian wolves in Northwest Spain. The glucocorticoid and sex hormone levels were measured in feces collected as a function of exposure (conspicuous/inconspicuous), height (above ground level/at ground level), and strategic location in the habitat (at crossroads/off crossroads), as well as the frequency of re-marking. The feces, believed to serve as marking cues, had higher glucocorticoid levels (cortisol) and sex hormones (testosterone, progesterone, and estradiol). The results suggest that in Iberian wolves, the alpha pair is subject to higher social stress than subordinate individuals, and that the reproductive suppression of subordinates is not mediated by chronic glucocorticoid elevation. PMID- 18392659 TI - Laparoscopic-assisted versus open abdominoperineal resection for low rectal cancer: a prospective randomized trial. AB - BACKGROUND: Laparoscopic resection of colonic cancer has been shown to improve postoperative recovery without jeopardizing tumor clearance and survival, but information on low rectal cancer is scarce. The aim of this randomized trial was to compare postoperative recovery between laparoscopic-assisted versus open abdominoperineal resection (APR) in patients with low rectal cancer. Recurrence and survival data were also recorded and compared between the two groups. METHODS: Between September 1994 and February 2005, 99 patients with low rectal cancer were randomized to receive either laparoscopic-assisted (51 patients) or conventional open (48 patients) APR. The median follow-up time of living patients was about 90 months for both groups. The primary and secondary endpoints of the study were postoperative recovery and survival, respectively. Data were analyzed by intention-to-treat principle. RESULTS: The demographic data of the two groups were comparable. Postoperative recovery was better after laparoscopic surgery, with earlier return of bowel function (P < .001) and mobilization (P = .005), and less analgesic requirement (P = .007). This was at the expense of longer operative time and higher direct cost. There were no differences in morbidity and operative mortality rates between the two groups. After curative resection, the probabilities of survival at 5 years of the laparoscopic-assisted and open groups were 75.2% and 76.5% respectively (P = .20). The respective probabilities of being disease-free were 78.1% and 73.6% (P = .55). CONCLUSIONS: Laparoscopic assisted APR improves postoperative recovery and seemingly does not jeopardize survival when compared with open surgery for low rectal cancer. A larger sample size is needed to fully assess oncological outcomes. PMID- 18392660 TI - Visceral obesity may affect oncologic outcome in patients with colorectal cancer. AB - PURPOSE: Obesity is closely related to the development of colorectal cancer as well as other metabolic complications. We investigated the prognostic significance of visceral obesity and body mass index (BMI) in 161 resectable colorectal cancer patients. METHODS: Ratios of visceral fat area (VFA) to subcutaneous fat area (SFA) were measured from the digital images of patients' computed tomography taken before the surgery, and patients were divided into those with high and those with low VFA/SFA ratio according to the degree of proportional visceral adiposity, and into an overweight and a normal-weight group according to their preoperative BMI. RESULTS: The overweight group showed a borderline decrease in cumulative disease-free survival compared to the normal weight group (P = 0.064). Patients with high VFA/SFA ratio (more than 50 percentiles) had significantly lower cumulative disease-free survival rate compared to patients with low VFA/SFA ratio (P = 0.008). BMI and visceral adiposity showed no influence on overall survival of patients. CONCLUSION: Increased visceral adiposity was a significant predictor of disease-free survival in patients with resectable colorectal cancer. The prognostic significance of visceral adiposity should further be determined in a larger set of patients. PMID- 18392661 TI - Differences in the pattern of presentation and treatment of proximal and distal gastric cancer: results of the 2001 gastric patient care evaluation. AB - BACKGROUND: While the overall incidence of gastric cancer has declined in the United States of America, the incidence of proximal gastric cancers has increased. The purpose of this analysis was to highlight key differences between proximal and distal gastric cancer as they relate to presentation and treatment. METHODS: Data on 6,099 patients diagnosed with gastric adenocarcinoma were collected as a patient care evaluation under the auspices of the American College of Surgeons Commission on Cancer. The chi-square (chi (2)) test was used for comparisons of proportions across levels of categorical variables by site. RESULTS: The proximal cancer group included 1,924 patients (87% cardia, 13% fundus) and the distal cancer group included 1,311 patients (85% antrum, 15% pylorus). Proportionately, proximal cancer cases were male (P < 0.01), younger (P < 0.01), and White (P < 0.01); whereas, distal gastric cancer cases were Black (P < 0.01), Hispanic (P < 0.01), and Asian (P = 0.01). Surgery alone (without adjuvant chemotherapy or radiation) was utilized more frequently in distal disease (39.5%) compared to proximal disease (25.7%) (P < 0.01). Preoperative adjuvant therapy was utilized more frequently in proximal disease (41.7%) compared to distal disease (2.1%) (P < 0.01). CONCLUSIONS: The populations that developed proximal verses distal gastric cancer differed with respect to sex, age, and racial background. Cancer-directed treatments also differed based upon tumor location. Understanding these differences may someday enable us to identify important high-risk populations, prevention strategies, and ultimately best treatment strategies. Long-term survival differences will be explored when follow up data become available. PMID- 18392662 TI - The future of health disparities research: 2008 and beyond. PMID- 18392663 TI - Sex hormones, their receptors and bone health. AB - Sex steroids regulate skeletal maturation and preservation in both men and women, as already recognized in the 1940s by Albright and Reifenstein. The impact of gonadal insufficiency on skeletal integrity has been widely recognized in adult men and women ever since. In the context of their skeletal actions, androgens and estrogens are no longer considered as just male and female hormones, respectively. Androgens can be converted into estrogens within the gonads and peripheral tissues and both are present in men and women, albeit in different concentrations. In the late 1980s, sex steroid receptors were discovered in bone cells. However, the understanding of sex steroid receptor activation and translation into biological skeletal actions is still incomplete. Due to the complex metabolism, sex steroids may have not only endocrine but also paracrine and/or autocrine actions. Also, circulating sex steroid concentrations do not necessarily reflect their biological activity due to strong binding to sex hormone binding globulin (SHBG). Finally, sex steroid signaling may include genomic and non-genomic effects in bone and non-bone cells. This review will focus on our current understanding of gonadal steroid metabolism, receptor activation, and their most relevant cellular and biological actions on bone. PMID- 18392665 TI - Ultrasonic monitoring of foamed polymeric tissue scaffold fabrication. AB - Polymeric tissue scaffolds are central to many regenerative medicine therapies offering a new approach to medicine. As the number of these regenerative therapies increases there is a pressing need for an improved understanding of the methods of scaffold fabrication. Of the many approaches to processing scaffolds, supercritical fluid fabrication methods have a distinct advantage over other techniques as they do not require the use of organic solvents, elevated processing temperatures or leaching processes. The work presented here is centred on the development of a new approach to monitoring supercritical scaffold fabrication based on determination of the scaffold acoustic impedance to inform protocols for scaffold fabrication. The approach taken uses an ultrasonic pulse echo reflectometer enabling non-invasive monitoring of the supercritical environment on-line. The feasibility of this approach was investigated for two scaffolds of different molecular weight. Acoustic results demonstrate that differences in the physical properties of the two scaffolds could be resolved, particularly during the foaming process which correlated with findings from time lapsed imaging and micro X-ray computed tomography (micro X-ray CT) images. Thus, this work demonstrates the feasibility of ultrasonic pulse-echo reflectometry to non-invasively study supercritical scaffold fabrication on-line providing a greater understanding of the scaffold fabrication process. PMID- 18392664 TI - Renal function and rate of hip bone loss in older men: the Osteoporotic Fractures in Men Study. AB - Older men with reduced renal function are at increased risk of hip bone loss. Given the robustness of this association across different measures and a growing body of literature, our findings indicate that clinicians should take into account renal function when evaluating older men for osteoporosis risk and bone loss. Future randomized controlled trials should test whether interventions in this high risk population are effective in preventing bone loss and decreasing fracture incidence. INTRODUCTION: Studies examining whether kidney impairment, not requiring dialysis, is associated with osteoporosis have reported conflicting results. METHODS: We tested the hypothesis that reduced renal function in older men as manifested by higher concentrations of cystatin C or lower levels of estimated glomerular filtration rate (eGFR) is associated with higher rates of bone loss. We measured serum cystatin C, serum creatinine and total hip bone mineral density (BMD) at baseline in a cohort of 404 older men enrolled in the Osteoporotic Fractures in Men (MrOS) Study and followed them prospectively for an average of 4.4 years for changes in BMD. Associations between renal function and change in hip BMD were examined using linear regression. RESULTS: In multivariable analysis, the mean rate of decline in total hip BMD showed an increase in magnitude with higher cystatin C concentration (mean annualized percent change -0.29, -0.34, -0.37 and -0.65% for quartiles 1 to 4; p for trend=0.004). Similarly, adjusted rates of hip bone loss were higher among men with lower eGFR as defined by the modification of diet in renal disease formula (mean annualized percent change -0.58, -0.39, -0.37, and -0.31 for quartiles 1 to 4; p for trend=0.02), but not among men with lower eGFR as defined by the Cockcroft-Gault formula (mean annualized percent change -0.47, -0.44, -0.31 and 0.43 for quartiles 1 to 4; p for trend=0.48). CONCLUSIONS: Older men with reduced renal function are at increased risk of hip bone loss. Our findings suggest that health care providers should consider renal function when evaluating older men for risk factors for bone loss and osteoporosis. PMID- 18392666 TI - Study on antibacterial mechanism of copper-bearing austenitic antibacterial stainless steel by atomic force microscopy. AB - A study was made on the antibacterial mechanism of copper-bearing austenitic antibacterial stainless steel by a series of methods such as atomic force microscopy (AFM) observation, force-distance curves and inductively coupled plasma mass spectrometer test. It was observed by AFM that the structure of the outer cell membrane responsible for the cell permeability was substantially changed for the bacteria after contacting with the antibacterial stainless steel, showing that cell walls were seriously damaged and a lot of contents in the cells leaked. It was also found that the adhesion force of bacteria to antibacterial stainless steel was considerably greater than that to the contrast steel, indicating that the electrostatic forces by Cu(2+ )being an important factor for killing bacteria. PMID- 18392667 TI - Preparation, characterization and mechanical performance of dense beta-TCP ceramics with/without magnesium substitution. AB - Beta-tricalcium phosphate (beta-TCP) powder was prepared by a two-step process: wet precipitation of apatitic tricalcium phosphate [Ca(9)(HPO(4))(PO(4))(5)(OH)] (beta-TCP 'precursor') and calcination of the precursor at 800 degrees C for 3 h to produce beta-TCP. Magnesium-substituted tricalcium phosphate (beta-TCMP) was produced by adding Mg(NO(3))(2) . 6H(2)O into Ca(NO(3))(2) solution as Mg(2+) source before the precipitation step. The transition temperature from beta-TCP to alpha-TCP increases with the increase of Mg(2+) content in beta-TCMP. beta-TCMP with 3 mol.% Mg(2+) has beta-TCP to alpha-TCP transition temperature above 1,300 degrees C. Dense beta-TCMP (3 mol.% Mg(2+)) ceramics ( approximately 99.4% relative density) were produced by pressing the green bodies at 100 MPa and further sintering at 1,250 degrees C for 2 h. The average compressive strength of dense beta-TCP ceramics sintered at 1,100 degrees C is approximately 540 MPa, while that of beta-TCMP (3 mol.% Mg(2+)) ceramics is approximately 430 MPa. PMID- 18392668 TI - Deposition of nano-hydroxyapatite particles utilising direct and transitional electrohydrodynamic processes. AB - Electrohydrodynamic spraying is a well established process used to deposit, coat, analyse and synthesise materials within the biomedical remit. Recently, electrohydrodynamic printing has been developed to afford structures for potential applications in the biomedical and medical engineering fields. Both of these processes rely on the formation of an electrically-induced jet, however the resulting products can be made strikingly different and offer potential in broader applications. Here we show how spraying and printing are linked by elucidating the ease of transition between the processes. Changes in the deposition distance can result in either spray (>10 mm) or print formation (<3 mm), with an overlap of the two in between this range. For the optimal printing distance of 0.5 mm, gradual changes in the applied voltage (0-4.5 kV) encounters transitional printing modes (dripping, micro-dripping, rapid micro-dripping, unstable and stable jetting) which can be utilised for patterning. The results indicate the robustness of the electrohydrodynamic route in the nano-materials processing arena, with emphasis on biomedical materials. PMID- 18392669 TI - The effect of sodium-ion implantation on the properties of titanium. AB - This paper deals with the surface modification of titanium by sodium-ion implantation and with the effect of this modification on structure, corrosion resistance, bioactivity and cytocompatibility. The Na ions were implanted with doses of 1 x 10(17) and 4 x 10(17) ions/cm(2) at an energy of 25 keV. The chemical composition of the surface layers formed during the implantation was examined by secondary-ion mass spectrometry (SIMS) and X-ray photoelectron spectroscopy (XPS), and their microstructure--by transmission electron microscopy (TEM). The corrosion resistance was determined by electrochemical methods in a simulated body fluid (SBF) at a temperature of 37 degrees C, after exposure in SBF for various times. The surfaces of the samples were examined by optical microscopy, by scanning electron microscopy (SEM-EDS), and by atomic force microscopy (AFM). Biocompatibility of the modified surface was evaluated in vitro in a culture of the MG-63 cell line and human osteoblast cells. The TEM results indicate that the surface layers formed during the implantation of Na-ions are amorphous. The results of the electrochemical examinations obtained for the Na implanted titanium samples indicate that the implantation increases corrosion resistance. Sodium-ion implantation improves bioactivity and does not reduce biocompatibility. PMID- 18392671 TI - So long as they grow out of it: comics, the discourse of developmental normalcy, and disability. AB - This essay draws on two emerging fields--the study of comics or graphic fiction, and disability studies--to demonstrate how graphic fictions articulate the embodied, ethical, and sociopolitical experiences of impairment and disability. Examining David B's Epileptic and Paul Karasik and Judy Karasik's The Ride Together, I argue that these graphic novels unsettle conventional notions of normalcy and disability. In so doing, they also challenge our assumed dimensions and possibilities of the comics genre and medium, demonstrating the great potential comics hold for disability studies. PMID- 18392670 TI - Risk factors for problematic gambling: a critical literature review. AB - This article is a critical review of risk factors for pathological gambling categorized by demographics, physiological and biological factors, cognitive distortions, comorbidity and concurrent symptoms, and personality symptoms and characteristics. There is also a varia section (availability, parents playing, sensory characteristics, schedules of reinforcement, age of onset, and playing duration). The review found very few well established risk factors for pathological gambling (i.e. more than two studies to support the conclusions). Well established risk factors included demographic variables (age, gender), cognitive distortions (erroneous perceptions, illusion of control), sensory characteristics, schedules of reinforcement, comorbid disorders (OCD, drug abuse), and delinquency/illegal acts. An understanding of risk factors for pathological gambling should enhance prevention and treatment approaches. PMID- 18392672 TI - Alcohol and sexual risk behavior among men who have sex with men in South african township communities. AB - There are few studies of HIV risk behavior among men who have sex with men (MSM) in sub-Saharan Africa. We conducted a behavioral survey of MSM in peri-urban "township" communities in Gauteng province, South Africa. Between October 2004 and March 2005, 199 MSM completed an interviewer administered pen-and-paper standardized questionnaire. The sample was 94% black, 83% identified as gay, and 12% as bisexual. Among those reporting sex with other men in the prior six months (n = 147), 59% reported unprotected anal intercourse (UAI). Increased risk of UAI was associated with both regular drinking (adjusted odds ratio (OR) 4.1, 95% confidence interval (CI) 1.4, 12.6) regular drinking to intoxication (OR 2.6, 95% CI 1.0, 6.8), and reporting symptoms of rectal trauma resulting from anal intercourse (AI; OR 4.3, 95% CI 1.8, 10.4). Decreased risk of UAI was associated with the exclusive use of lubricants that are latex-compatible for AI (OR 0.3, 95% CI 0.1, 0.7). Township MSM in South Africa are at high risk of HIV infection. Targeted sexual health and risk reduction interventions that address the link between alcohol use and HIV risk are urgently needed. PMID- 18392673 TI - Use of time-location sampling for systematic behavioral surveillance of truck drivers in Brazil. AB - We assessed HIV-related risk behavior in a time-location sampling (TLS) interviewer administered survey of male truck drivers at the crossroads of two of the major highways used for trucking in Northeast Brazil (N = 686). Overall, 21.3% had sex while on the road (2.8% unprotected); 12.3% had sex with commercial sex workers (1.4% unprotected); 1.8% had sex with hitch-hikers (0.3% unprotected). Unprotected sex with non-wife partners while on the road was independently associated with stimulant use, lack of religious affiliation and being separated or divorced. Stimulant use may be a particular occupational hazard associated with risk for HIV among truck drivers in Brazil and should be considered as a point of intervention. The TLS sampling approach can form the basis of systematic behavioral surveillance in this highly mobile population. PMID- 18392674 TI - Male age influences oocyte-donor program results. AB - BACKGROUND: The purpose of our research was to examine the relationship between male age and semen parameters in a range of ages (from 20s to 60s) in Egg Donation Program (EDP) cycles. EDP provides a pool of high quality oocytes, thus allowing better analysis of the sperm efficacy. DISCUSSION: The retrospective study population consisted of 484 male partners of patients undergoing EDP in in vitro fertilization (IVF) treatments. A comparison was made of male age and sperm parameters within two groups: cycles resulting in a pregnancy (pregnant group) and cycles which failed to achieve a pregnancy (non-pregnant group). The men involved in the pregnant group were found to be significantly younger 43.2 +/- 8.1 than those of the nonpregnant group 46.81 +/- 7.8 (p = 0.003). Analysis of sperm morphology revealed a significant prevalence of teratozoospermia in males of the non-pregnant group, as compared to the males of the pregnant group (29% vs. 11%, respectively). The results also demonstrate that sperm parameters are apparently not diminished until men reach the age of 40. However, between 40-50 years old semen parameters deteriorate. Male age was found to be related to a reduction in sperm strict criteria: 44.8 in normozoospermia, 47.9 (p = 0.02), 48.4 (p = 0.04) and 51.9 (p = 0.001) years old in mild teratozoospermia, moderate teratozoospermia and severe teratozoospermia, respectively. Additionally, the results showed that the percentage of "healthy" embryos on day 3 of embryo culture was lower in the non-pregnant group (26%), as compared with the pregnant group (34%; p = 0.01). CONCLUSION: Our study confirms that sperm parameters are reduced by age and suggests that this age-dependent effect could be a reason for failures in IVF cycles even in EDP couples. PMID- 18392675 TI - Growth hormone co-treatment within a GnRH agonist long protocol in patients with poor ovarian response: a prospective, randomized, clinical trial. AB - OBJECTIVE: The efficacy of growth hormone co-stimulation to long luteal GnRHa regimen in poor responders to COH for IVF was assessed. METHODS: This prospective, randomized, clinical trial was performed in a private assisted reproduction center. The study involved 61 patients who responded poorly to high dose gonadotropin treatment in their first cycles in the same center. Study group of 31 patients were given growth hormone co-treatment, daily subcutaneous injection of 4 mg from day 21 of preceding cycle along with GnRHa, until the day of hCG. Control group of 30 patients received the same treatment protocol except the growth hormone cotreatment. Primary end-point of the study was the number of oocytes fertilized whereas the pregnancy rate was the secondary end-point. RESULTS: Patients' demographic characteristics did not differ significantly between the two groups. 2PNs in growth hormone co-treatment group was significantly higher than the control group (4.4 +/- 1.8 vs 1.5 +/- 0.9, p < 0.001). Although more pregnancies and more clinical pregnancies with fetal heart beat were achieved in growth hormone group (12/31), compared to the control group (6/30), the difference did not reach to statistical significance. CONCLUSION: Poor responder women undergoing repeated assisted reproduction treatment and co stimulated with GH achieve more oocytes, higher fertilization rate if growth hormone started in the luteal phase of previous cycle, as compared with women of the same status treated with GnRHa long protocol. The study was unable to show that clinical pregnancy rate was increased significantly. PMID- 18392676 TI - Association of progesterone receptor polymorphisms with recurrent implantation failure after in vitro fertilization and embryo transfer. AB - INTRODUCTION: Progesterone is the hormone of pregnancy and is required for its initiation. The actions of progesterone are mediated by the progesterone receptor. Polymorphic variants of human progesterone receptor genes have been implicated in implantation failure. MATERIALS AND METHODS: We, therefore, investigated the prevalence of H770H (C/T genotype), V660L polymorphism and a 306 bp Alu insertion in exon 7 of the progesterone receptor among women with history of recurrent implantation failure to determine whether any of these polymorphisms may serve as a risk factor for implantation failure. DNA was extracted from the buccal swabs obtained from 66 women experiencing implantation failure and 75 fertile control women. PCR amplification of fragments was purified and the DNA sequenced to identify the polymorphism. The frequencies for the three variants were 27% for H770H, 21% for V660L and 0% for the 306 bp Alu insertion in exon 7 among women with implantation failure compared with control women of 25% for H770H and 24%for V660L and 0% for the 306 bp Alu insertion in exon 7. DISCUSSION: No significant differences in the overall allelic frequency of progesterone receptor variants was seen when women experiencing recurrent implantation failure were compared with control women. CONCLUSION: We conclude that the H770H and V660L and PROGINS progesterone receptor polymorphisms are not markers that can identify women at risk for recurrent implantation after IVF/ET. PMID- 18392677 TI - Toxicological effects of military fog oil obscurant on Daphnia magna and Ceriodaphnia dubia in field and laboratory exposures. AB - Our purpose was to determine if the acute and sub-lethal effects of fog oil, an obscurant used for military training, could be observed in realistic field exposures. To this end, we exposed Daphnia magna to oil fogs under actual release conditions at a U.S. Army training site. Guided by field investigations, acute toxicity experiments were conducted in the laboratory with the more sensitive species Ceriodaphnia dubia to test the hypothesis that dissolution of fog oil constituents into water is minimal and actual contact by organisms with the water surface is required to cause toxicity. We conducted further experiments to test the hypothesis that vaporization of fog oil alters its chemical composition and toxicity to freshwater invertebrates. In the field, daphnid mortality was minimal more than 5 m from the point of fog generation, but sub-lethal effects were more extensive. Both field and laboratory experiments suggested that physical contact with oils on the water surface was the most important factor driving toxicity. To our knowledge, this is the first attempt to evaluate toxicological endpoints with freshwater invertebrates in field exposures with fog oil. PMID- 18392678 TI - Inpatient admissions for drug-induced liver injury: results from a single center. AB - Objective To review all cases of drug-induced liver injury (DILI) requiring hospitalization at a single tertiary care center. Methods Patient records were identified by ICD-9 codes for inpatient visits from November 1998 through March 2006. Results Of a total 83,265 hospital admissions during the study period, 40 were for DILI (0.048%). Thirteen patients had non-acetaminophen DILI (NA-DILI); 27 had acetaminophen-related DILI (A-DILI). In the NA-DILI group, mean age was 59 +/- 17.9 years and liver injury was classified as hepatocellular (7), cholestatic (5), or mixed (1). A variety of medications were implicated with antimicrobials being the most common class. Resolution occurred in seven, two died of complications related to hepatotoxicity, one underwent liver transplantation, and the outcome was undetermined in three who were lost to follow-up. In the A-DILI group, mean age was 35 +/- 11.0 years. Eighteen involved intentional overdose of acetaminophen; nine were associated with chronic use. The pattern of injury was hepatocellular in all. Resolution occurred in 4 patients, death in 8, and improvement in 15. Conclusions DILI is a rare cause of inpatient admission but is associated with significant mortality. Spontaneous resolution occurs in most patients but return to normal liver function may take months. Antimicrobial agents account for the largest proportion of NA-DILI. PMID- 18392680 TI - Multi-molecular gradients of permissive and inhibitory cues direct neurite outgrowth. AB - Correct development of neuronal tracts requires the coordination of multiple permissive and inhibitory signals. By generating an in vitro microenvironment using soft lithography and microfluidic techniques, multiple guidance cues can be presented in a spatially defined way. Here we evaluated how neurites of dorsal root ganglia neurons responded to permissive and inhibitory cues presented by substrate-bound molecular gradients. Linear gradients containing inhibitory chondroitin sulfate proteoglycan (CSPG) and/or permissive laminin-1 (LN) were generated as single-cue gradients, parallel double-cue gradients, and opposing double-cue gradients with varying slopes. Neurite growth was analyzed using circular statistical methods, and for all gradients examined, neurons extended neurites toward regions of lower CSPG and higher LN concentrations. Single-cue gradients elicited similarly directed neurite growth responses at the higher concentrations tested for both LN and CSPG, and both gradient slope and fractional concentration change affected neurite growth. When the two contrasting molecular cues were presented together, neurites responded differently depending on the directions of the gradients. Neurite growth on LN-CSPG double gradients of opposite direction was strongly directed, while neurite growth on LN-CSPG double gradients of parallel direction was uniform. These results represent an important step toward understanding how neurite growth is guided by complex microenvironments containing multiple molecular cues. PMID- 18392679 TI - Atorvastatin does not improve liver biochemistries or Mayo Risk Score in primary biliary cirrhosis. AB - Statin treatment reduces hypercholesterolemia and may be anti-inflammatory. Case reports noted decreased alkaline phosphatase and histological improvement following statin treatment in primary biliary cirrhosis. The objective of this study was to assess the long-term effects of statin treatment in primary biliary cirrhosis. A retrospective analysis compared clinical and biochemical data from 15 hypercholesterolemic individuals with primary biliary cirrhosis who were treated long-term with atorvastatin with an age and gender matched, primary biliary cirrhosis control group. A significant decrease in total cholesterol and low-density lipoprotein (LDL)-cholesterol (p < or = 0.002) was observed throughout atorvastatin treatment (median time 2.5 years). LDL-cholesterol levels in the control group were not significantly changed after 2 years (p > 0.050). No significant changes were noted in alanine aminotransferase (ALT), alkaline phosphatase, total bilirubin and Mayo Risk Score in either group (p > 0.05). Long term atorvastatin treatment reduced LDL-cholesterol in primary biliary cirrhosis, but there was no evidence of any anti-inflammatory effect. PMID- 18392681 TI - The death of ouabain-treated renal epithelial cells: evidence against anoikis occurrence. AB - The mechanisms of cell death signaling triggered by cardiotonic steroids are poorly understood. Based on massive detachment of ouabain-treated Madin-Darby canine kidney (MDCK) cells, it may be proposed that the cytotoxic action of these compounds is mediated by anoikis, i.e. a particular mode of death occurring in cells lacking cell-to-extracellular matrix interactions. We tested this hypothesis. Six hour incubation of MDCK cells with ouabain, marinobufagenin or K+ free medium almost completely blocked Na+,K+-ATPase, increased Na (i) + content by approximately 10-fold and suppressed cell attachment to regular-plastic-plates by up to 5-fold. In contrast, the death of attached cells was observed after 24-h incubation with ouabain but not in the presence of marinobufagenin or K+-free medium. Cells treated with ouabain and undergoing anoikis on ultra-low attachment plates exhibited different cell volume behaviour, i.e. swelling and shrinkage, respectively. The pan-caspase inhibitor z-VAD.fmk and the protein kinase C activator PMA rescued MDCK cells from anoikis but did not influence the survival of ouabain-treated cells, whereas medium acidification from pH 7.2 to 6.7 almost completely abolished the cytotoxic action of ouabain, but did not significantly affect anoikis. Our results show that the Na (i) + ,K (i) + -independent mode of MDCK cell death evoked by ouabain is not mediated by anoikis. PMID- 18392682 TI - Trifolin acetate-induced cell death in human leukemia cells is dependent on caspase-6 and activates the MAPK pathway. AB - In the present study we demonstrated that the flavonoid derivative trifolin acetate (TA), obtained by acetylation of naturally occurring trifolin, induces apoptosis. Associated downstream signaling events were also investigated. TA induced cell death was prevented by the non-specific caspase inhibitor z-VAD-fmk and reduced by the presence of the selective caspase inhibitors z-LEHD-fmk (caspase-9), z-DEVD-fmk (caspase-3) and z-VEID-fmk (caspase-6). The apoptotic effect of TA was associated with (i) the release of cytochrome c from mitochondria which was not accompanied by dissipation of the mitochondrial membrane potential (DeltaPsi(m)), (ii) the activation of the mitogen-activated protein kinases (MAPKs) pathway and (iii) abrogated by the over-expression of Bcl 2 or Bcl-x(L). TA-induced cell death was attenuated by inhibition of extracellular signal-regulated kinases (ERK) 1/2 with U0126 and inhibition of p38(MAPK) with SB203580. In contrast, inhibition of c-Jun NH(2)-terminal kinase (JNK) by SP600125 significantly enhanced apoptosis. Although reactive oxygen species (ROS) increased in response to TA, this did not seem to play a pivotal role in the apoptotic process since different anti-oxidants were unable to provide cell protection. The present study demonstrates that TA-induced cell death is mediated by an intrinsic-dependent apoptotic event involving mitochondria and MAPK, and through a mechanism independent of ROS generation. PMID- 18392683 TI - Prolactin induces MFG-E8 production in macrophages via transcription factor C/EBPbeta-dependent pathway. AB - The lactogenic hormone prolactin (PRL) regulates milk protein gene expression in mammary glands. To maintain homeostatic balance in the body, milk fat globule epidermal growth factor 8 (MFG-E8) is vital for phagocytic clearance of apoptotic cells. We investigated the effects of PRL on MFG-E8 expression in macrophages by evaluating its promoter function. Macrophages were stimulated with PRL, and the expression of MFG-E8 was determined using real-time PCR and Western blotting. The role of MFG-E8 on phagocytosis of apoptotic cells in PRL-treated macrophages was assessed using microscopy, while the response of PRL to MFG-E8 expression was evaluated using luciferase assay. Following treatment with PRL, significant up regulations of the PRL receptor and MFG-E8 were observed in macrophages, though PRL-treated macrophages more efficiently engulfed apoptotic cells. The results of MFG-E8 promoter analysis showed considerable up-regulation of promoter activity in macrophages following PRL treatment and results from mutation analysis of the MFG-E8 promoter suggested that the C/EBPbeta binding site was responsible for PRL induced activation of the MFG-E8 promoter. C/EBPbeta activity was found to be up regulated in PRL-treated cells as revealed by an electrophoretic mobility shift assay (EMSA). In conclusion, PRL is a potent inducer of MFG-E8 expression in macrophages, while its effect is mediated by the presence of a responsive element in the MFG-E8 promoter. PMID- 18392684 TI - Current therapeutic options for breast cancer central nervous system metastases. AB - OPINION STATEMENT: Breast cancer metastases to the central nervous system (CNS) has devastating consequences for the individual. As treatment options for metastatic breast cancer expand and as quality of life and overall survival improve, researchers are targeting potential treatments for this sanctuary site. Attention is now being focused on defining the phenotype of breast cancer that has a propensity to metastasize to the CNS. Specific therapies that penetrate the blood brain barrier as well as adjuvant therapies that decrease recurrence in the CNS are currently being investigated. We will review current approaches to the diagnosis, evaluation, and treatment of CNS metastases in breast cancer patients. PMID- 18392685 TI - Thaxtomin biosynthesis: the path to plant pathogenicity in the genus Streptomyces. AB - Streptomyces species are best known for their ability to produce a wide array of medically and agriculturally important secondary metabolites. However, there is a growing number of species which, like Streptomyces scabies, can function as plant pathogens and cause scab disease on economically important crops such as potato. All of these species produce the phytotoxin thaxtomin, a nitrated dipeptide which inhibits cellulose synthesis in expanding plant tissue. The biosynthesis of thaxtomin involves conserved non-ribosomal peptide synthetases, P450 monooxygenases, and a nitric oxide synthase, the latter being required for nitration of the toxin. This nitric oxide synthase is also responsible for the production of diffusible nitric oxide by scab-causing streptomycetes at the host pathogen interface, suggesting that nitric oxide production might play an additional role during the infection process. The thaxtomin biosynthetic genes are transcriptionally regulated by an AraC/XylS family regulator, TxtR, which is conserved in pathogenic streptomycetes and is encoded within the thaxtomin biosynthetic gene cluster. The TxtR protein specifically binds cellobiose, a known inducer of thaxtomin biosynthesis, and cellobiose is required for expression of the biosynthetic genes. A second virulence gene in pathogenic Streptomyces species, nec1, encodes a novel secreted protein that may suppress plant defence responses. The thaxtomin biosynthetic genes and nec1 are contained on a large mobilizable pathogenicity island; the transfer of this island to recipient streptomycetes likely explains the rapid emergence of new pathogenic species. The newly available genome sequence of S. scabies will provide further insight into the mechanisms utilized by pathogenic streptomycetes during plant microbe interactions. PMID- 18392686 TI - Auto-immune neutropenia associated with metformin in a patient with splenic marginal zone lymphoma. AB - A 75-year-old man with splenic marginal zone lymphoma had been treated by fludarabine. The 3 first courses were very well tolerated, with no cytopenia. To treat diabetes mellitus, metformin was added at day 6 after the beginning of the fourth course of fludarabine treatment. He had previously been briefly treated with metformin a few months before. Four days later, at day 10, haemogram revealed severe neutropenia (neutrophils: 0.7 g/l). Neutrophil count continued to decrease reaching 0.283 g/l at day 17. A bone marrow aspirate was performed and showed very active granulopoiesis with maturation arrest at the promyelocyte stage. Metformin was discontinued on day 22 (16 days after reintroduction) and neutropenia promptly reversed during next week with 1.9 g/l (day 29). Direct and indirect granulocyte immunofluorescence test were used to detect anti-neutrophil autoantibodies. The antibodies were related to metformin and were detectable by granulocyte agglutination test and the monoclonal antibody-specific immobilization of granulocyte antigens assay. A possible explanation is the fludarabine-metformin combination, since fludarabine could deplete regulator T lymphocytes and lead to the emergence of metformin-induced anti-neutrophil antibodies. PMID- 18392687 TI - Understanding, testimony and interpretation in psychiatric diagnosis. AB - Psychiatric diagnosis depends, centrally, on the transmission of patients' knowledge of their experiences and symptoms to clinicians by testimony. In the case of non-native speakers, the need for linguistic interpretation raises significant practical problems. But determining the best practical approach depends on determining the best underlying model of both testimony and knowledge itself. Internalist models of knowledge have been influential since Descartes. But they cannot account for testimony. Since knowledge by testimony is possible, and forms the basis of psychiatric diagnosis, its very existence is a factor in support of an externalist model of knowledge in general. Internalist and externalist models of knowledge also suggest different ways of responding to the practical challenges of basing psychiatric diagnosis on testimony. Thus the argument in favour of externalism also supports a potentially empirically testable hypothesis about interpretation of non-native speakers for accurate psychiatric diagnosis: interpretation of non-English speakers should be as transparent and unhindered by specialised medical knowledge as possible. PMID- 18392688 TI - Health-related quality of life following blind rehabilitation. AB - PURPOSE: The purpose of this study was to investigate the effect of residential blind rehabilitation on patients' vision-targeted health-related quality of life (HRQOL) and general physical and mental function. METHOD: The National Eye Institute 25-item Visual Function Questionnaire (NEI VFQ) plus appendix questions, the 12-item Short-Form Health Survey (SF-12), Hope Scale and Coopersmith self-esteem inventory were administered to 206 legally blind veterans prior to their entering a residential (in-patient) blind rehabilitation program and again to 185 and 176 of the original cohort at 2 and 6 months after completion of the rehabilitation program, respectively. Data on visual acuity, visual field extent, contrast sensitivity and scanning ability were also collected. The duration of the in-patient rehabilitation programs ranged from 11 to 109 days. Questionnaire scores were compared pre-rehabilitation and post rehabilitation. RESULTS: Following rehabilitation there was a significant improvement in nine of 11 NEI VFQ subscales and in a composite score at both the 2- and 6-month post-rehabilitation intervals. Mental health (SF-12) and self esteem also improved significantly although physical health ratings declined over the course of the study (approximately 10 months). CONCLUSIONS: Residential blind rehabilitation appears to improve patients' self-reported vision-targeted HRQOL, self-esteem and mental health aspects of generic HRQOL. PMID- 18392689 TI - Immunotherapy for head and neck cancer. AB - Head and neck cancer represents a challenging disease. Despite recent treatment advances, which have improved functional outcomes, the long-term survival of head and neck cancer patients has remained unchanged for the past 25 years. One of the goals of adjuvant cancer therapy is to eradicate local regional microscopic and micrometastatic disease with minimal toxicity to surrounding normal cells. In this respect, antigen-specific immunotherapy is an attractive therapeutic approach. With the advances in molecular genetics and fundamental immunology, antigen-specific immunotherapy is being actively explored using DNA, bacterial vector, viral vector, peptide, protein, dendritic cell, and tumor-cell based vaccines. Early phase clinical trials have demonstrated the safety and feasibility of these novel therapies and the emphasis is now shifting towards the development of strategies, which can increase the potency of these vaccines. As the field of immunotherapy matures and as our understanding of the complex interaction between tumor and host develops, we get closer to realizing the potential of immunotherapy as an adjunctive method to control head and neck cancer and improve long-term survival in this patient population. PMID- 18392691 TI - Reviewing stem cells: a new day. PMID- 18392690 TI - Arterial elasticity and plasma levels of adiponectin and leptin in type 2 diabetic patients treated with thiazolidinediones. AB - BACKGROUND: Thiazolidinediones (TZDs) improve peripheral insulin sensitivity, but the effect on arterial stiffness is less clear. The aim of the present study was to assess the differential effect of pioglitazone or rosiglitazone on arterial stiffness and plasma levels of adiponectin and leptin in patients with type 2 diabetes mellitus. METHODS: Thirty-five type 2 diabetic subjects were randomly assigned to receive pioglitazone (30 mg/day; n = 14), rosiglitazone (4 mg/day; n = 11), or placebo (medical nutrition therapy; n = 10) for 12 weeks. Changes in plasma glucose, glycosylated hemoglobin, insulin resistance (HOMA-IR), total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, adiponectin, and leptin were evaluated at baseline and after 12 weeks. In parallel, large arterial compliance (C1) and small arterial compliance (C2) were measured at baseline and at the end of treatment period. RESULTS: At 12 weeks, the rosiglitazone (P = 0.026) and pioglitazone (P = 0.004) groups had a significant increase from baseline in adiponectin that was not seen in the medical nutrition therapy group. No significant changes in plasma leptin and in C1 and C2 elasticity indexes were observed over the entire study period in any of the treatment groups. CONCLUSIONS: In this study of patients with type 2 diabetes, treatment with TZDs was associated with a significant improvement in adiponectin levels, although no significant effects were seen on leptin levels and arterial elasticity. PMID- 18392692 TI - Lung cancer patients with previous or simultaneous urologic cancers. AB - STUDY OBJECTIVES: There have been few reports on clinical characteristics of lung cancer patients with previous or simultaneous urologic cancers. Additionally, it was scarcely reported whether these patients could tolerate standard therapy. To evaluate them, we conducted a retrospective study. METHODS: The records of patients with lung cancer who had previously or simultaneously urological cancers seen in our division between January 1985 and August 2007 were reviewed. RESULTS: During the study period, 1,105 patients with lung cancer were seen at our division. Thirteen (1.2%) had previous or simultaneous urological cancers (11 males). Eleven patients were smokers. The diagnosis of urological cancers preceded the diagnosis of lung cancer in all but three patients, for whom the diagnosis of urologic cancers was made during the workup of primary lung cancer. Histologically, the lung cancers included six squamous cell carcinomas, four adenocarcinomas, and three small cell lung cancers. Four underwent surgical resection. Six were treated with platinum-containing chemotherapy, but none of them developed severe renal toxicity. The median survival following the diagnosis of lung carcinoma for NSCLC patients was 18 months, and for SCLC it was 24 months. The cause of death in the 12 patients who died in the study period was directly related to lung cancer, and recurrence of urological cancers was observed in none of the patients. CONCLUSIONS: For patients with urological cancers, smoking cessation, a chest radiograph or CT scan at least yearly and swift evaluation of signs or symptoms that are suggestive of lung cancer should be recommended. PMID- 18392693 TI - A method for quantifying the unstable and highly polar drug nafamostat mesilate in human plasma with optimized solid-phase extraction and ESI-MS detection: more accurate evaluation for pharmacokinetic study. AB - An advanced quantification method was developed with solid-phase extraction (SPE) and mass spectrometry (MS) determination for nafamostat, an unstable and highly polar drug, in human plasma. For unstable drugs with an ester group, the main analytical challenge is how to avoid the ester hydrolysis, and strong acid or alkaline conditions should be excluded during sample preparation. Considering that, we developed a relatively mild method with SPE for sample preparation without strong acid and alkaline treatment, which was optimized with different pHs and salt concentrations in phosphate-buffered saline treatment. The results indicated that pH 5 gave the most efficient extraction and 0.1 M salt concentration enhanced the extraction the most, with a minor effect on MS monitoring. The extraction method effectively avoided drug hydrolysis and achieved good drug enrichment over 82.2%. The linear range of quantification was 1.25-160 ng mL(-1). The stability of the drug in sample treatment was fully validated according to the sample processing procedure, including the stability in fresh blood, mobile phase, plasma and acidic methanol, and the results indicated that the drug remained stable during the whole sample preparation. Compared with a previous isotope-labeling method, more accurate and specific quantification of plasma concentration was achieved with liquid chromatography electrospray ionization MS determination. With use of our method, nafamostat mesilate pharmacokinetics in 30 Chinese healthy volunteers was investigated with three doses via intravenous-drip infusion. The pharmacokinetic parameters were also estimated and compared with those of Japanese volunteers (slightly lower plasma concentration and longer terminal elimination half-life for Chinese volunteers). The difference in the pharmacokinetics may be ascribed to the quantification method, because previous isotope labeling may have overestimated the parent drug. PMID- 18392694 TI - Specific leaf area relates to the differences in leaf construction cost, photosynthesis, nitrogen allocation, and use efficiencies between invasive and noninvasive alien congeners. AB - Comparisons between invasive and native species may not characterize the traits of invasive species, as native species might be invasive elsewhere if they were introduced. In this study, invasive Oxalis corymbosa and Peperomia pellucida were compared with their respective noninvasive alien congeners. We hypothesized that the invasive species have higher specific leaf (SLA) than their respective noninvasive alien congeners, and analyzed the physiological and ecological consequences of the higher SLA. Higher SLA was indeed the most important trait for the two invaders, which was associated with their lower leaf construction cost, higher nitrogen (N) allocation to photosynthesis and photosynthetic N use efficiency (PNUE). The higher N allocation to photosynthesis of the invaders in turn increased their PNUE, N content in photosynthesis, biochemical capacity for photosynthesis, and therefore light-saturated photosynthetic rate. The above resource capture-, use- and growth-related traits may facilitate the two invaders' invasion, while further comparative studies on a wider range of invasive and noninvasive congeners are needed to understand the generality of this pattern and to fully assess the competitive advantages afforded by these traits. PMID- 18392695 TI - Deep venous thrombosis and previous myocardial infarction in mild factor XII deficiency: a risk factor for both venous and arterial thrombosis. AB - Factor XII deficiency is associated with increased risk for both arterial and venous thrombosis. We describe a case of DVT involving superficial femoral and popliteal vein occurred following total hip replacement and despite prophylaxis with low molecular weight heparin in a subject with previous acute myocardial infarction (AMI). Tests of haemostasis documented a slightly prolonged activated partial thromboplastin time (APTT) (45'') due to mild factor XII deficiency (clotting activity 32%). A therapeutic dose of enoxaparin was started, together with warfarin therapy. The patient was advised to continue oral anticoagulation indefinitely. Although cases of both venous and arterial thrombosis in carriers of severe factor XII deficiency have been already reported, to our knowledge this is the first case in the literature occurred in a carrier of partial factor XII deficiency. In conclusion, factor XII deficiency should be suspected if a patient presents with recurrent arterial and/or venous thrombosis and prolonged APTT. If this defect is diagnosed, in the presence of a history of thrombotic events, lifelong anticoagulation could be considered. PMID- 18392696 TI - Mammary tumor development from T47-D human breast cancer cells in obese ovariectomized mice with and without estradiol supplements. AB - Obesity is a risk factor for postmenopausal breast cancer, particularly for development of estrogen-receptor (ER)-positive tumors. Additionally, obesity is implicated in breast cancer progression. However, few studies address mechanisms of action of how obesity mediates these responses. Our goal was to address how obesity and/or elevated serum leptin affects tumor formation from ER-positive T47 D cells. In Study 1 ovariectomized CD-1 nude female mice were injected with goldthioglucose (GTG) at 0.5 mg/g body weight in saline or the vehicle at 6 weeks of age. At 10 weeks of age mice were inoculated with T47-D cells and implanted with estrogen pellets. In Study 2 mice were injected with 0.3 mg/g GTG or the vehicle. At 10 weeks of age cells were inoculated and mice were implanted with estrogen or placebo pellets. Mice were followed until 30 weeks of age. Some GTG mice became obese and others were non-responders. In Study 1 no mice developed tumors. In Study 2 mice with placebo pellets developed more tumors than mice with estrogen pellets, 50% vs. 13%. GTG-obese mice with placebo pellets had a 100% tumor incidence compared to 50% and 20% for GTG-lean and controls without estrogen. Serum leptin was higher in obese compared to lean mice and adiponectin was not affected by body weight. Adiponectin:leptin ratio was significantly reduced in obese compared to lean mice. Leptin, leptin receptor and signaling protein expression were determined in mammary and tumor tissue. Leptin and STAT3 were most abundant in tumors. These findings suggest that in vivo estrogen suppressed proliferation of T47-D cells but without supplemental estrogen obesity enhanced tumor development. The exact reason for this is not presently clear. PMID- 18392697 TI - Overexpression of the cucumber LEAFY homolog CFL and hormone treatments alter flower development in gloxinia (Sinningia speciosa). AB - Leafy (LFY) and LFY-like genes control the initiation of floral meristems and regulate MADS-box genes in higher plants. The Cucumber-FLO-LFY (CFL) gene, a LFY homolog in Cucumis sativus L. is expressed in the primordia, floral primordia, and each whirl of floral organs during the early stage of flower development. In this study, functions of CFL in flower development were investigated by overexpressing the CFL gene in gloxinia (Sinningia speciosa). Our results show that constitutive CFL overexpression significantly promote early flowering without gibberellin (GA(3)) supplement, suggesting that CFL can serve functionally as a LFY homolog in gloxinia. Moreover, GA(3) and abscisic acid (ABA) treatments could modulate the expression of MADS-box genes in opposite directions. GA(3) resembles the overexpression of CFL in the expression of MADS box genes and the regeneration of floral buds, but ABA inhibits the expression of MADS-box genes and flower development. These results suggest that CFL and downstream MADS-box genes involved in flower development are regulated by GA(3) and ABA. PMID- 18392698 TI - [Depression: still an underestimated disease]. PMID- 18392700 TI - Donor evaluation and hepatectomy for living-donor liver transplantation. AB - In the past decade, considerable technical advances have been accomplished in living-donor liver transplantation (LDLT). The procedure has become accepted globally as a standard modality for the treatment of end-stage liver disease and hepatocellular carcinoma in both pediatric and adult populations. During the period of this procedure's development, however, tragedy has occurred. Serious morbidity and even mortality have been experienced and reported in live donors. The transplant community has been very much aware of its responsibilities toward live donor care, and much effort has been made to improve and secure the overall outcomes of donors. Unlike in deceased-donor liver transplantation (DDLT), opportunity or chance plays a lesser role in the availability of an organ for LDLT. Judgment calls are often made by individuals; therefore, the evaluation process includes social and ethical aspects not encountered among the usual indications for hepatobiliary surgical disease. Thus, the selection of live donors should be made from a wider perspective compared with that for conventional patient care. The approach to selecting live donors may vary slightly between the West, where a large number of DDLTs are performed daily, and the Far East, where they are much less frequent. However, the recognition that the transplant community has a responsibility to provide care to living donors is common. This review provides an overview of the current donor evaluation and surgical procedures involved in LDLT, with the recognition that an open and educated debate is key to ensuring public confidence and maintaining ethical standards in the field. PMID- 18392701 TI - The use of marginal grafts in liver transplantation. AB - Because of the shortage of organ supplies, more transplant programs have begun to use marginal grafts in liver transplantation. A number of single-center experiences with marginal grafts have yielded encouraging results, but recent analyses using nationwide databases show that outcomes are inferior to results with normal whole-liver grafts. Use of marginal grafts is still acceptable, however, and plays an important role in expanding the donor pool and decreasing mortality on the waiting list. In the broadest terms, national data and single center experiences show that: (1) there is no limit in donor age for liver transplantation, (2) appropriate selection of steatotic livers improves outcomes, (3) prolonged graft ischemia is a preventable factor, (4) livers from donors with hepatitis B or C virus can be safely transplanted, and (5) adequate prophylaxis prevents recurrence of hepatitis B without significant graft loss. In addition, grafts procured after cardiac death are another growing source of marginal grafts. Transmission of malignancy from donors is rare but life-threatening. Reduced-size grafts from living-donor or split-liver transplantation have shown similar outcomes to whole-liver transplantation. In this review, we will discuss the current status of the utility of these marginal grafts in liver transplantation. PMID- 18392702 TI - Strategies for improving the outcomes of small-for-size grafts in adult-to-adult living-donor liver transplantation. AB - Living-donor liver transplantation (LDLT) has been refined and accepted as a valuable treatment for patients with end-stage liver disease in order to overcome the shortage of organs and mortality on the waiting list. However, graft size problems, especially small-for-size (SFS) grafts, remain the greatest limiting factor for the expansion of LDLT, especially in adult-to-adult transplantation. Various attempts have been made to overcome the problems regarding SFS grafts, such as increasing the graft liver volume and/or controlling excessive portal inflow to a small graft, with considerable positive outcomes. Recent innovations in basic studies have also contributed to the treatment of SFS syndrome. Herein, we review the literature and assess our current knowledge of the pathogenesis and treatment strategies for the use of SFS grafts in adult-to-adult LDLT. PMID- 18392703 TI - Treatment strategy for hepatitis C after liver transplantation. AB - A significant proportion of patients with chronic hepatitis C virus (HCV) infection develop liver cirrhosis and complications of end-stage liver disease over periods of two to three decades and require liver transplantation, although re-infection is common and leads to further adverse events, given that such patients receive life-long immunosuppression. Because of the critical organ shortage worldwide, living-donor liver transplantation has an important role in many countries, especially in the Far East. Despite previous arguments, the results of recent well-designed studies suggest equivalent outcomes for deceased donor and living-donor liver transplantation. No specific immunosuppression regimen has proven advantageous, but the general rule is "low and slow". Combined pegylated interferon and ribavirin therapy is the current standard treatment, but compared to this therapy in the immunocompetent population, its efficacy in clearing the virus remains low. Moreover, its general application is hindered by the high prevalence of intolerability, lowering its efficacy from the aspect of intention to treat. Retransplantation becomes an option when treatment for disease progression fails, but it should be considered at an earlier stage in patients with a lower MELD (model for end-stage liver disease) score compared to that used for primary transplantation, which is a great challenge with the current critical organ shortage. The need for new anti-HCV drugs to further delay disease progression or even to enhance viral clearance, presumably specific HCV life-cycle inhibitors, is urgent. The liver transplant community must maintain an open mind regarding the development of these drugs and focus on their availability for early clinical trials in liver transplant recipients. PMID- 18392704 TI - Liver transplantation for hepatocellular carcinoma. AB - The role of liver transplantation (LT) in patients with hepatocellular carcinoma (HCC) has evolved over the past two decades, and transplantation has become one of the few curative treatment modalities for patients with HCC. Early results were poor, but the current restrictive selection criteria can yield excellent results. This review will discuss recent issues in the field, including (1) factors affecting the recurrence of HCC after LT; (2) the effect of downstaging HCC before LT, including transarterial catheter chemoembolization (TACE) and radiofrequency ablation (RFA); and (3) living-donor versus deceased-donor liver transplantation for HCC patients. The most important factors that have been described to affect LT survival include the tumor size, vascular invasion, and the degree of tumor differentiation. Recently, tumor markers, including alpha fetoprotein and des-gamma carboxy prothrombin, were reported as predictors of HCC recurrence after LT. Furthermore, the experience accumulated with locoregional therapies such as TACE and RFA as bridging procedures to LT, along with the reduced waiting time under the HCC-adjusted MELD (model for endstage liver disease) system for organ allocation has led to improved outcomes. With the recent advances in adult living-donor liver transplantation (LDLT), there may be a marked change in the role of liver transplantation for hepatic malignancies, in particular for HCC. PMID- 18392705 TI - Current status of liver transplantation across ABO blood-type barrier. AB - Outcomes of ABO-blood type incompatible liver transplantation have recently improved owing to various treatments. The typical clinical manifestations of antibody mediated rejection (AMR) are hepatic necrosis and intrahepatic biliary complication (IHBC). The prognosis of AMR is poor. AMR is the result of circulatory disturbance which is caused by injury to the endothelium due to an antibody-antigen-complement reaction. Diffuse C4d staining in the portal capillaries and periportal areas in severe AMR. Since natural antibodies against A/B carbohydrate determinants are likely to develop as a result of exposure to environmental bacteria that express similar determinants, the B-1 lineage has been speculated to be the major population of B-cell types responding to A/B determinants. Calcineurin inhibitors block B-1 cell differentiation. Rituximab can be used to deplete both cells that are producing IgM antibodies and those that have already differentiated into B-1 cells. Mycophenolate mofetil is required to inhibit the production of IgG subclass of antibodies. The outcome is now similar to that of blood-type-matched transplantation. However, there are still issues to be solved in order to further improve the outcome via a decrease of infection. PMID- 18392706 TI - Domino liver transplantation. AB - Orthotopic liver transplantation is today an established treatment for end stage liver diseases. However, the ongoing shortage of suitable livers together with progressively longer waiting lists prevents many patients from being transplanted, and many patients die while being on the waiting list. Using livers from living donors is one way to increase the supply of liver grafts. Another group of potential living liver donors are some selected liver recipients, whose native explanted liver in turn can be considered for transplantation into another patient. This unorthodox procedure have been named domino liver transplantation (DLT). The domino approach can be considered in patients with some genetic or biochemical disorders that today are treated by liver transplantation. The underlying rationale is that such livers ultimately cause severe systemic disease but are otherwise normal. In this review we present the current world status of DLT as well as updated results from the Domino Liver World Transplant Register (DLTR) and our own experience at the Karolinska University Hospital Huddinge with the DLT procedure. PMID- 18392707 TI - Duodenum-preserving total pancreatic head resection for cystic neoplastic lesions in the head of the pancreas. AB - BACKGROUND/PURPOSE: Cystic neoplastic lesions of the pancreas are now found with increasing frequency. Duodenum-preserving pancreatic head resection with segmental resection of the duodenum has been introduced for the surgical treatment of inflammatory and neoplastic lesions. We report the following data from 15 patients treated surgically for cystic neoplastic lesions of the pancreas head. METHODS: Duodenum-preserving total pancreatic head resection (DPPHRt) with segmental resection of the duodenum (SD) was performed in eight patients, five with intraductal papillary mucinous neoplasm (IPMN), two with mucinous cystic neoplasm (MCN), and one with cystic endocrine neoplasm (EN). In four patients, a subtotal pancreatic head resection was performed, but recurrence of the IPMN lesion was observed in two patients. Ten patients suffered cystadenoma, three patients had a borderline lesion, and two patients had an in-situ carcinoma. RESULTS: Eight patients had a DPPHRt with SD resection, two patients had a resection of the uncinate process including segmental resection of the inferior duodenal segment, and one patient had a duodenum-and spleen-preserving total pancreatectomy. In four patients a DPPHR with subtotal pancreatic head resection was carried out. Postoperative local complications occurred in eight patients: there was a recurrence of the IPMN lesion in the remnant pancreatic head in two patients; and there was intraabdominal bleeding in one patient, pancreatic fistula in one patient, and delay of gastric emptying in four patients. Seven patients showed signs of acute pancreatitis. Hospital mortality was 0%, and postoperative length of hospital stay was 10. 4 days (range, 8-18 days). CONCLUSIONS: Duodenum-preserving total pancreatic head resection for IPMN, MCN, serous cystadenoma (SCA), and cystic EN lesions is a safe and beneficial surgical procedure. Segmental resection of the duodenum was applied for an oncologically complete resection. In regard to long-term outcome, the procedure is, additionally, a pancreatic cancer preventive strategy. PMID- 18392708 TI - Gallbladder carcinoma: intraoperative imprint cytology, a helpful and valuable screening procedure. AB - BACKGROUND/PURPOSE: A major problem arises when a tumor is not readily recognized at the time of cholecystectomy. A carcinoma at early stages or one hidden by acute or chronic inflammation can be overlooked during surgery, and the diagnosis would then be made only after microscopic examination of paraffin-embedded tissue. The purpose of the present report is to communicate the results of 10 years' experience with the use of imprint cytology for the intraoperative diagnosis of gallbladder carcinoma. METHODS: During the period June 1994 to June 2004, 525 imprint cytologies of gallbladder mucosa were performed after 160 open cholecystectomies and 365 laparoscopic cholecystectomies. The patients had been operated on for acute or chronic cholecystitis due to lithiasis. Only 18 patients had a preoperative diagnosis of gallbladder carcinoma. The sensitivity, specificity, predictive values, accuracy, and their confidence intervals (95% CIs), of intraoperative imprint cytology for the diagnosis of carcinoma were analyzed. RESULTS: The average time employed for each procedure was 10 min. Patients' mean age was 69.2 years with a range of 24 to 92 years. Three hundred and forty-two patients (65.1%) were women and 183 (34.9%) were men. The imprint cytology method was positive in 44 of 58 gallbladder carcinomas diagnosed, with a sensitivity of 75.9% (44/58 patients) and a specificity of 99.8% (466/467). Only 1 case (1/525; 0.2%) was a false-positive; this was due to reactive changes. The positive predictive value was 97.8% (44/45; 95% CI = 86.8-99.9), negative predictive value was 97.1% (466/480; 95% CI = 95.0-98.3), and accuracy was 97.1% ([44 + 466]/525; 95% CI = 95.2-98.3). There was insufficient or inadequate material in 4 cases. CONCLUSIONS: Imprint cytology of the gallbladder mucosa is an easy, rapid, and high-quality method for detecting gallbladder carcinoma. PMID- 18392709 TI - Does ampullary carcinoma arise from distended glands in the papilla of Vater? AB - BACKGROUND/PURPOSE: Glands near the surface of the papilla of Vater may become distended with mucus and become hyperplastic; that is, become distended. In this study, we tried to clarify carcinogenesis in these distended glands. METHODS: Twenty-nine pancreatoduodenectomized nontumorous duodenal papilla specimens from carcinoma of the pancreas and bile duct and 34 resected ampullary carcinoma specimens were studied histopathologically and immunohistochemically, using cytokeratins and mucin immunohistochemical features/phenotypes. RESULTS: Distended glands were found in 11 of the 29 pancreatoduodenectomized specimens. These glands were immunopositive for cytokeratin (CK) 7 and MUC-5AC Glycoprotein (MUC5AC), but not for CK20, while the intrapapillary portion was CK7-positive and CK20-negative, but mostly negative for MUC5AC. Immunopositivity for CK7, CK20, and MUC5AC was found in 25, 21, and 18 of the 34 specimens of ampullary carcinoma, respectively. In 23 of the 34 specimens, immunoreactivity for MUC5AC and that for CK7 was coincident, that is, when the former was immunopositive, so was the latter, and vice versa, while in 25 of the 34 specimens, immunoreactivity for MUC5AC was opposite to that for CK20. Among the 23 cases in which immunoreactivity for MUC5AC and CK7 was coincident, 10 were MUC5AC+, CK7+, CK20- and 7 were MUC5AC-, CK7-, CK20+, suggestive of disease arising from the pancreaticobiliary mucosa or the distended glands in the former and disease arising from the duodenal mucosa in the latter. In MUC5AC+ cases, other than the 10 cases of MUC5AC+, CK7+, CK20-, 6 were double-positive and 1 was double negative for CK7 and CK20, and 1 was CK20-positive, and at least 1 case showing double-negativity for CK7 and CK20 was suggestive of disease arising from the distended glands. CONCLUSIONS: Although most ampullary carcinomas arise from the duodenal mucosa or intra-ampullary mucosa, both CK7-positive and MUC5AC-positive or only MUC5ACpositive ampullary carcinomas may arise from the distended glands. PMID- 18392710 TI - Characterization of K-ras gene mutations in association with mucinous hypersecretion in intraductal papillary-mucinous neoplasms. AB - BACKGROUND/PURPOSE: Intraductal papillary-mucinous neoplasms (IPMNs) of the pancreas have a favorable prognosis. However, invasive ductal carcinomas of the pancreas show a rapid progression. The aim of this study was to investigate gene mutations in pure pancreatic juice from IPMN patients and to define these genetic mutations in relation to the histopathological and clinical features of IPMNs. METHODS: Twenty-two patients with IPMN, 21 patients with ductal carcinoma, and 20 patients with normal pancreas or chronic pancreatitis were recruited for this study. We measured the main pancreatic duct's largest diameter and the maximum size of a dilated branch was assessed by ultrasonography or endoscopic ultrasonography. Pure pancreatic juice was collected and was investigated for K ras, p16, and p53 mutations. RESULTS: Mutant K-ras gene was detected in 13 of the 22 patients (59.1%) with IPMNs. Different kinds of mutations were detected in the same patient in 4 cases. In the 13 patients with mutant K-ras gene, the diameter of the most dilated part of the main pancreatic duct was 2-8 mm (average, 4.5 mm) and in 7 patients with wild-type K-ras gene, the diameter was 2-5 mm (average, 2.7 mm). There was a significant difference in the diameter of the main pancreatic duct between patients with and without the mutant K-ras gene (P = 0.0323). CONCLUSIONS: The incidence of K-ras mutation may be associated with the hypersecretion of mucin. PMID- 18392711 TI - Long-term results of elective hepatectomy for the treatment of ruptured hepatocellular carcinoma. AB - BACKGROUND/PURPOSE: According to the General rules for the clinical and pathological study of primary liver cancer, compiled by the Liver Cancer Study Group of Japan, ruptured hepatocellular carcinoma (HCC) is classified as stage 4, even if the tumor is small and solitary. We examined the long-term results of elective hepatectomy for the treatment of ruptured HCC. METHODS: A first hepatectomy was performed without operative death in 193 patients with HCC. Ten patients had ruptured HCC (ruptured group) and 183 patients had nonruptured HCC (nonruptured group). The extension of HCC was macroscopically classified as stage 1 in 23 patients, stage 2 in 71, stage 3 in 53, and stage 4 in 46. RESULTS: Cumulative survival rates in the ruptured group at 1, 5, and 10 years were 90.0%, 67.5%, and 20.3%, respectively. The cumulative survival rate was lower in patients with stage 4 disease in the nonruptured group than that in patients in the ruptured group (P < 0.05). Cumulative survival rates did not differ significantly between patients in the ruptured group and those with stage 2 or stage 3 disease. CONCLUSIONS: Survival rates after elective hepatectomy in patients with ruptured HCC are good, even if the disease is classified as stage 4. PMID- 18392712 TI - Intraductal papillary mucinous neoplasm of the pancreas: clinical characteristics and treatment outcomes of 118 consecutive patients from a single center. AB - BACKGROUND/PURPOSE: Appropriate surgical treatment strategies based on clinicopathological findings are unavailable for intraductal papillary mucinous neoplasm (IPMN) of the pancreas. We investigated the clinical features of pancreatic IPMN in a single-center database in order to design an optimal surgical strategy. METHODS: The medical records of 118 consecutive patients who had undergone surgical resection between August 1994 and December 2004, in whom IPMN was histologically confirmed, were reviewed retrospectively for radiological and pathological findings. RESULTS: Most of the invasive carcinomas in these patients were detected as the main-duct type (88.5%). The type of tumor (main duct type vs branched-duct type), the tumor size, and the dilated duct size were significant predictive factors associated with malignancy. The relative risk of malignancy was greatest at 13-mm or more ductal dilation in the main-duct type (Odds ratio, 4.1), at 35-mm or more tumor size (Odds ratio, 7.6), and for main duct type (Odds ratio, 3.9). Major pancreatic resections such as total pancreatectomy and pancreatoduodenectomy were performed in 14.5% and 69% of the patients, respectively. There was a 19.5% rate of incomplete resection, with these patients having a positive resection margin. However, significant recurrence did not occur in patients with a benign IPMN lesion which remained at the resection margin. The overall postoperative survival rate at 5 years was 98.2% for benign IPMN and 65.3% for malignant IPMN. CONCLUSIONS: Function preserving strategies, based on the clinical status of the patient, are necessary in order to avoid possible severe metabolic complications following extended pancreatectomy in patients with benign IPMN because of the low recurrence rate and good prognosis of this entity, irrespective of margin status. PMID- 18392713 TI - Detection of circulating tumor cells in patients with pancreatic cancer: a preliminary result. AB - BACKGROUND/PURPOSE: It has been reported that circulating tumor cells (CTCs) can be used to predict survival in metastatic breast cancer. In this preliminary study, we examined the level of CTCs in pancreatic cancer (PC) patients to elucidate whether we could predict survival in PC. METHODS: The eligible subjects, at Tokyo Medical University Hospital, were 26 patients with PC, 11 with chronic pancreatitis, and 10 healthy volunteers. Three PC patients underwent surgery, 18 patients (who were stage IV) were treated with gemcitabine (GEM), and 5 patients received best supportive care (BSC). RESULTS: The CTC count was 1/7.5 ml blood or higher (defined as positive) in 11 of the 26 patients (42%; mean, 16.9/7.5 ml blood; range, 1-105/7.5 ml blood). Gemcitabine was administered to 6 of the 11 CTC-positive patients (3.8 courses on average). The treatment was continued for more than three courses in 2 patients, in both of whom the CTC count was only 1/7.5 ml blood. Operation was performed in 1 of the 11 CTC positive patients. The remaining 4 patients of the 11 CTC-positive patients received only BSC. CTC was negative in 15 patients with PC (stage II, 1; stage III, 1; stage IVa, 7; and stage IVb, 6) and in the subjects with benign conditions. The median survival times (MSTs) of the CTC-positive and-negative patients were 110.5 and 375.8 days (P < 0.001). When the analysis was limited to the 14 stage-IVb patients, the MSTs of the CTC-positive and-negative patients were 52.5 and 308.3 days (P < 0.01). CONCLUSIONS: The present study demonstrated that the detection of CTCs in peripheral blood may be useful to predict prognosis in patients with PC. PMID- 18392714 TI - Follicular lymphoma of the extrahepatic bile duct mimicking cholangiocarcinoma. AB - We report a case of a 33-year old man who presented with symptoms and signs of an acute biliary tract obstruction with jaundice and abdominal pain. Diagnostic imaging studies revealed a biliary stricture of the hepatic confluence, and a mass at the hepatic hilum which obstructed the extrahepatic bile duct from the outside. At laparotomy, there was 3-cm-size nodule at the hilum which presented with a rubbery consistency. We performed extrahepatic bile duct resection and right and left hepaticojejunostomy. Histological examination of the resected specimen revealed follicular lymphoma, which consisted of medium cleaved follicle like cells, grade 1 of 3 according to the revised European-American classification of lymphoid neoplasms proposed by International Lymphoma Study Group. Postoperative follow-up of more than 1 year has been completely uneventful, without any symptoms or signs of disease recurrence. This is the second case report of follicular lymphoma of the extrahepatic bile duct. PMID- 18392715 TI - Perioperative management of benign hepatic tumors in patients with glycogen storage disease type Ia. AB - Glycogen storage disease type Ia (GSD-Ia; von Gierke disease) is an inherited disorder caused by glucose-6-phosphatase deficiency, and there have been some reports of hepatic tumors in patients with this disease. We report two patients with benign hepatic tumors with GSD-Ia. One is a 19-year-old man who underwent segmentectomy 4 for a focal nodular hyperplasia, and the other is a 31-year-old woman who underwent segmentectomies 3, 5, and 6 for hepatic adenomas. Two significant perioperative complications, resulting from the carbohydrate metabolic disorders, hypoglycemia and metabolic acidosis, occurred in both patients. We managed the metabolic complications successfully by administering a sufficient volume of glucose intravenously. Close perioperative monitoring of blood glucose and lactate concentrations is essential in the perioperative management of patients with GSD-Ia. The intravenous administration of glucose, starting with a smaller dose and then increasing the dose, is adequate management for lactic acidosis with or without hypoglycemia during the perioperative period. PMID- 18392716 TI - Patient with perforation caused by emphysematous cholecystitis who showed flare on the skin of the right dorsal lumbar region and intraperitoneal free gas. AB - We report an 84-year-old man with perforation caused by emphysematous cholecystitis who showed flare on the skin of the right dorsal lumbar region and intraperitoneal free gas. The patient was admitted for abdominal pain, abdominal swelling, and consciousness disorder 18 days after the onset. Abdominal computed tomography (CT) revealed emphysema in the gallbladder and a small amount of intraperitoneal free gas. Intraoperative findings suggested gangrenous cholecystitis. The gallbladder wall was perforated, and an abscess involving the right subphrenic region, the periphery of the liver and gallbladder, and the right paracolonic groove, was detected. The flare on the body surface may have reflected abscess formation in the right abdominal cavity. Emphysematous cholecystitis induces necrosis and perforation in many patients, and immediate strategies such as emergency surgery are important. PMID- 18392717 TI - Renal cell carcinoma with unusual metastasis to the gallbladder. AB - Gallbladder involvement in patients with renal cell carcinoma (RCC) is extremely rare. We present a report of a 61-year-old man with a synchronous RCC metastasis to the gallbladder presenting as an intraluminal polypoid mass simulating primary gallbladder carcinoma. Enhanced abdominal computed tomography demonstrated a well enhanced polypoid lesion in the gallbladder. Intraoperative rapid pathological examination of the gallbladder tumor showed clear cell-type cancerous cells. Microscopically, tumor cells of both the resected kidney and gallbladder had round uniform nuclei, clear cytoplasm, and well-defined cytoplasmic borders, forming alveolar patterns. Immunohistochemically, the tumor cells were negative for cytokeratin 7 (CK7) and carcinoembryonic antigen (CEA), which is usually positive in primary clear cell carcinoma of the gallbladder. Therefore, the final diagnosis was RCC with a synchronous gallbladder metastasis. PMID- 18392718 TI - Metastasis of hepatocellular carcinoma to spleen and small intestine. AB - Metastasis of cancer to the spleen or small intestine is rare. We encountered a case of hepatocellular carcinoma (HCC) with splenic and jejunal metastases. A 60 year-old man was referred to us in February 2005 with a diagnosis of splenic tumor. Since 2001, he had been treated repeatedly for HCC with liver cirrhosis due to hepatitis C virus infection; partial liver resection, transcatheter arterial chemo-embolization, and radiofrequency ablation therapy had been performed. In October 2004, he had undergone partial pulmonary resection due to metastasis of HCC to the lung. The splenic tumor, which was detected by computed tomography, seemed to be a metastasis of HCC. Splenectomy was performed for the splenic tumor, and a jejunal tumor was discovered and also resected. Both the splenic and jejunal tumors were diagnosed pathologically as metastases from the HCC. After repeated treatment for HCC, metastases can appear in various organs; thus, careful observation is necessary during follow-up. PMID- 18392719 TI - Gallbladder agenesis with a stone in the cystic duct bud. AB - An 84-year-old woman was admitted to the hospital because of pyloric stenosis caused by gastric cancer. Abdominal computed tomography and magnetic resonance imaging failed to demonstrate the gallbladder, but showed a gallstone in a duct like structure parallel to the common bile duct. When laparotomy was performed, the gallbladder and the fossa were not observed, and a blind-end duct, similar to a cystic duct, was found beside the common bile duct. Incisional exploration of the common bile duct was done after distal gastrectomy; the gallstone was not found in the common bile duct, but in the duct parallel to it. By observing the duct beneath the common bile duct with a cholangioscope, we considered it to be a hypoplastic cystic duct. After the gallstone was removed, a T-tube was placed into the common bile duct. Agenesis of the gallbladder is a rare congenital anomaly and is often asymptomatic. As far as we know, this is the first report of gallbladder agenesis with a hypoplastic cystic duct impacted with a stone. Careful intraoperative examination using a cholangioscope is useful to confirm the structure of the common bile duct. PMID- 18392720 TI - Mixed hepato/cholangiocarcinoma with paraneoplastic hypercalcemia. AB - Malignant hypercalcemia without bone metastasis may result from the abnormal secretion of parathyroid hormone (PTH) or PTH-related protein (PTH-rP). We present a case of possible PTH-rP-secreting mixed hepato/cholangiocarcinoma. In July 2000, a 52-year-old woman presented with right hypochondrial pain and weight loss. Imaging procedures revealed a 10-cm lesion involving the right hepatic lobe and the hilum. Retrohepatic inferior vena caval invasion was detected by abdominal angiography. Serum calcium was elevated, while intact PTH was undetectable. The rising level of hypercalcemia rapidly became life-threatening. Medical treatment with diphosphonate and octreotide was ineffective. Chemoembolization of the right branch of the hepatic artery, preceded by right portal vein embolization, helped regain normocalcemia and improve the patient's general condition. Extended right hepatectomy with prosthetic replacement of the vena cava and resection of the left portal vein was performed. Histopathology revealed mixed hepato/cholangiocarcinoma. Hypocalcemia appeared postoperatively but was successfully corrected. Recurrence of diffuse hepatic nodules resulted in recurrent hypercalcemia and death 3 and a half years after the surgery. This is the third case to be reported in the English-language literature. Paraneoplastic PTH-rP-induced hypercalcemia, which may be a fatal complication, can respond to arterial chemoembolization. Extensive hepatic resection may offer a good chance for survival. PMID- 18392721 TI - Hepatic venous outflow obstruction after right lateral sector living-donor liver transplantation, treated by insertion of an expandable metallic stent. AB - Hepatic venous outflow obstruction is a relatively uncommon but important and devastating complication occurring after liver transplantation. Recently, right lateral sector liver grafts have sometimes been used in living-donor liver transplantation (LDLT), but, to our knowledge, early hepatic venous outflow obstruction has never been reported in right lateral sector LDLT. A 58-year-old woman was diagnosed with liver cirrhosis and hepatocellular carcinoma and underwent right lateral sector LDLT. Postoperatively, she developed liver dysfunction. Doppler ultrasound examination revealed flat waveforms and low-flow velocity in the right hepatic vein (RHV). A computed tomography (CT) scan revealed a ventrally distorted RHV due to hypertrophy of the liver graft. Hepatic venous obstruction was suspected and a hepatic venogram was performed. The venogram revealed stenosis of the RHV due to the distortion of the vein. We performed percutaneous transfemoral balloon dilatation, but this was not effective. We then inserted an expandable metallic stent (EMS) into the RHV. After the EMS placement, the condition of the patient improved. Venogram and CT data suggested that the obstruction of the RHV developed because of distortion of the RHV to the ventral side during liver regeneration. PMID- 18392722 TI - Two-stage operation for hepatocellular carcinoma with synchronous bilateral adrenal gland metastasis. AB - Hepatocellular carcinoma (HCC) frequently metastasizes to the adrenal glands. The surgical strategy for HCC associated with bilateral adrenal gland metastasis is complicated because of the possibility of both postoperative hepatic failure and adrenal insufficiency. We herein report a patient with HCC with synchronous bilateral adrenal gland metastasis that was treated successfully with a two-stage operation. A 58-year-old man with HCC (12 cm in diameter) in the right lobe of the liver and bilateral adrenal gland tumors (right, 4 cm in diameter; left, 5 cm in diameter) was admitted to our hospital. Extended right hemihepatectomy and right adrenalectomy were performed simultaneously. The postoperative course was uneventful. Three months after this operation, left adrenalectomy was performed via a retroperitoneal approach. Hydrocortisone supplement was given, and the postoperative course was again uneventful. No recurrence was observed during the 10-month follow-up period. Two-stage surgery is a safe treatment option for giant HCC with synchronous bilateral adrenal gland metastasis. PMID- 18392723 TI - Carcinomatous meningitis secondary to cholangiocarcinoma without other systemic metastasis. AB - Carcinomatous meningitis is clinically less common than brain metastasis or spinal cord compression, but has dire consequences for both the quality of life and the overall survival of patients with various kinds of malignancies. It occurs in about 5% of all adult cancer patients, though autopsies may double this number. The primary tumors that frequently cause carcinomatous meningitis include lung cancer, breast cancer, leukemia, lymphoma, and melanoma. Carcinomatous meningitis secondary to primary tumors in the gastrointestinal tract is clinically infrequent. In this report, we describe a 73-year-old man with lower bile duct cancer, who developed carcinomatous meningitis following surgical resection of the primary cancer. To our knowledge, this is the third case of carcinomatous meningitis secondary to cholangiocarcinoma described in the literature so far. PMID- 18392724 TI - Resected xanthogranulomatous pancreatitis. AB - Xanthogranulomatous changes in the pancreas are extremely rare. A 66-year-old man presented with a 2-year history of epigastralgia. Computed tomography scan revealed a 4-cm low-density area around the body of the pancreas. Magnetic resonance imaging demonstrated that the mass appeared hyperintense on a T2 weighted image and isointense on a T1-weighted image. Based on a diagnosis of invasive ductal carcinoma of the pancreas, distal pancreatectomy and splenectomy were performed. Sections examined from the mass showed an aggregation of many foamy histiocytes, lymphocytes, and plasma cells. The surrounding pancreatic tissue showed fibrosis and chronic inflammation. These findings suggested a xanthogranulomatous inflammation, and resulted in a diagnosis of xanthogranulomatous pancreatitis. PMID- 18392725 TI - Histopathological effects of hexavalent chromium in the ovary of a fresh water fish, Channa punctatus (Bloch). AB - The histopathological effects of hexavalent chromium (Cr VI) in the ovary of a fresh water teleost, Channa punctatus were investigated. An exposure-dependent alteration in ovarian histology is reported. For both acute and chronic exposures to Cr (VI), the percentages of atretic oocytes were increased; this increase was more pronounced in the acute exposure group. A decrease in percentage of vitellogenic oocytes was observed in the chronic exposure group indicating impairment of vitellogenesis. The hepatocellular vacuolization and atrophy along with pyknotic nuclei in both acute and chronic chromium exposed fish liver supports the vitellogenic impairment. The observed alterations may be due to both direct cytotoxic effect of Cr (VI) on the ovary as well as mediation by overall systemic toxicity affecting other vital organs. PMID- 18392726 TI - Larvicidal activity of saponin from Achyranthes aspera against Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae). AB - The acetone, chloroform, ethyl acetate, hexane and methanol leaf extracts of Acalypha indica, Achyranthes aspera, Leucas aspera, Morinda tinctoria and Ocimum sanctum were studied against the early fourth-instar larvae of Aedes aegypti L and Culex quinquefasciatus Say. The larval mortality was observed after 24 h exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in the ethyl acetate extract of A. aspera. In the present study, bioassay-guided fractionation of A. aspera led to the separation and identification ofa saponin as a potential mosquito larvicidal compound, with LC50 value of 18.20 and 27.24 ppm against A. aegypti and C. quinquefasciatus, respectively. 1H NMR, 13C NMR and mass spectral data confirmed the identification of the active compound. This is the first report on the mosquito larvicidal activity of the saponin from the ethyl acetate extract of A. aspera. This study investigates the potential of crude extracts from commonly used medical herbs in India as an environmentally safe measure to control the vector of dengue and lymphatic filariasis. PMID- 18392727 TI - Methanolic extracts of Uncaria rhynchophylla induce cytotoxicity and apoptosis in HT-29 human colon carcinoma cells. AB - In this paper, we report the anticancer activities of Uncaria rhynchophylla extracts, a Rubiaceae plant native to China. Traditionally, Uncaria rhynchophylla has been used in the prevention and treatment of neurotoxicity. However, the cytotoxic activity of Uncaria rhynchophylla against human colon carcinoma cells has not, until now, been elucidated. We found that the methanolic extract of Uncaria rhynchophylla (URE) have cytotoxic effects on HT-29 cells. The URE showed highly cytotoxic effects via the MTT reduction assay, LDH release assay, and colony formation assay. As expected, URE inhibited the growth of HT-29 cells in a dose-dependent manner. In particular, the methanolic URE of the 500 microg/ml showed 15.8% inhibition against growth of HT-29 cells. It induced characteristic apoptotic effects in HT-29 cells, including chromatin condensation and sharking occurring 24 h when the cells were treated at a concentration of the 500 microg/ml. The activation of caspase-3 and the specific proteolytic cleavage of poly (ADP-ribose) polymerase were detected over the course of apoptosis induction. These results indicate that URE contains bioactive materials with strong activity, and is a potential chemotherapeutic agent candidate against HT 29 human colon carcinoma cells. PMID- 18392728 TI - A practical device for pinpoint delivery of molecules into multiple neurons in culture. AB - We have developed a device for pinpoint delivery of chemicals, proteins, and nucleic acids into cultured cells. The principle underlying the technique is the flow of molecules from the culture medium into cells through a rupture in the plasma membrane made by a needle puncture. DNA transfection is achieved by stabbing the needle tip into the nucleus. The CellBee device can be attached to any inverted microscope, and molecular delivery can be coupled with conventional live cell imaging. Because the position of the needle relative to the targeted cultured cells is computer-controlled, efficient delivery of molecules such as rhodamine into as many as 100 HeLa cells can be completed in 10 min. Moreover, specific target cells within a single dish can be transfected with multiple DNA constructs by simple changes of culture medium containing different plasmids. In addition, the nano-sized needle tip enables gentle molecular delivery, minimizing cell damage. This method permits DNA transfection into specific hippocampal neurons without disturbing neuronal circuitry established in culture. PMID- 18392729 TI - GABAC receptor subunit mRNA expression in the rat superior colliculus is regulated by calcium channels, neurotrophins, and GABAC receptor activity. AB - The distribution of mRNA for the rho2 subunit of the GABA(C) receptor is much broader in organotypic SC cultures than in vivo, suggesting that GABA(C) receptor expression is regulated by environmental factors. Electrophysiological recordings indicate that neurons in SC cultures have functional GABA(C) receptors, although these receptors exhibited smaller conductance than in vivo, probably due to increased rho2 subunit expression. Adding cortical input, treatment with various neuromodulators, and blocking neuronal activity with TTX failed to affect the expression of rho2 subunits. Electrophysiological recordings revealed the presence of spontaneous Ca(2+) currents in SC cultures and preventing these, by treatment with blockers of L-type Ca(2+) channels, caused rho2 mRNA expression to decline to in vivo levels. In contrast, rho1 subunit mRNA levels remained unchanged, indicating that the two subunits are independently regulated. Surprisingly, both tonic activation and blockade of GABA(C) receptors upregulated rho1/rho2 mRNA expression. Further, NGF and BDNF promoted such expression during an early postnatal time window. In vivo, expression of the rho2 mRNA in the SC, and the rho2/rho3 mRNA in the retina increased with age. Expression of the rho2 mRNA in the visual cortex, and the rho1 mRNA in the retina and SC was constant. Subunit mRNA expression was similar in dark-reared animals, indicating that visual experience has no influence. These experiments suggest that GABA(C) receptor expression in the SC is regulated during postnatal development. While visual experience seems to have no influence on GABA(C) receptor subunits, spontaneous calcium currents selectively promote rho2 expression and both rho1 and rho2 are autoregulated both by GABA(C) receptor activity and by neurotrophic factors. PMID- 18392730 TI - Differential activation of extracellular signal-regulated kinase 1 and a related complex in neuronal nuclei. AB - The extracellular signal-regulated kinases 1 and 2 (ERKs 1/2) are known to participate in regulating transcription in response to moderate depolarization, such as synaptic stimulation, but how the same active enzyme can differentially regulate distinct transcriptional programs induced with abnormal depolarization (high potassium) is unknown. We hypothesized that ERK1 or 2 accomplishes this differential nuclear response through close association with other proteins in stable complexes. In support of this hypothesis, we have found that immunoreactivity for an apparent high molecular weight complex containing phospho ERK1 increased in response to synaptic stimulation, but decreased in response to high potassium; p-ERK immunoreactivity at 44/42 kDa increased in both cases. Evidence supporting the conclusion that the band of interest contained ERK1 in a complex, as opposed to it being an unrelated protein crossreacting with antibodies against p-ERK, is that ERK1 (p44 MAPK) and 14-3-3 protein were electroeluted from the 160-kDa band cut from a gel. We also found the nuclear complexes to be exceptionally durable, suggesting a role for the crosslinking enzyme, transglutaminase, in its stabilization. In addition, we found other components of the ERK pathway, including MEK, ERK2, p90RSK, and Elk-1, migrating at higher-than-expected weights in brain nuclei. These results describe a novel stable complex of ERK1 in neuronal nuclei that responds differentially to synaptic and depolarizing stimulation, and thus may be capable of mediating gene transcription in a way distinct from the monomeric protein. PMID- 18392731 TI - Distribution of the scaffolding proteins PSD-95, PSD-93, and SAP97 in isolated PSDs. AB - We compared the distribution of three scaffolding proteins, all belonging to a family of membrane-associated guanylate kinases, thought to have key roles in the organization of the postsynaptic density (PSD). Isolated PSDs readily adhered to treated glass coverslips where they were labeled with immunogold and rotary shadowed for analysis by EM. The distribution of proteins within individual PSDs were measured by counting and mapping individual immunogold particles. PSD-95, as previously described, is distributed evenly throughout the PSD. We find here that PSD-93 has a nearly identical distribution suggesting that PSD-95 and PSD-93 could perform similar roles. SAP97, in contrast, is concentrated near edges of cleft sides of the PSDs, and in small clumps on their cytoplasmic sides. The homogenous distribution of PSD-95 and PSD-93 throughout the PSD is consistent with their being part of a backbone that stabilizes their various binding partners within the PSD. The distribution of SAP97 confirms that this protein is actually an integral component of the PSD, and suggests that it may have a role in inserting or stabilizing its main binding partner, Glu-R1, at the edge of the PSD. PMID- 18392732 TI - Practical evaluation of the drug-related problem management process in Swiss community pharmacies. AB - OBJECTIVE: To develop and evaluate a coding system integrated into pharmaceutical software to routinely report and assess the process of community pharmacists' interventions related to medical prescriptions. SETTING: A convenient sample of 20 Swiss community pharmacies. METHOD: Pharmacists documented their interventions concerning all drug-related problems (DRPs) related to medical prescriptions during four consecutive weeks in 2005. The coding system assesses each step of the DRP management process; that is, the type of problem, possible negative outcomes, pharmaceutical decisions, and individuals involved. In order to be comprehensive, the management process of technical problems related to prescriptions and clinical DRPs was analysed separately. MAIN OUTCOME MEASURE: DRP intervention rate and characterization of each step of the process. RESULTS: Of 38,663 prescriptions, 287 clinical DRPs required interventions. This corresponds to a mean intervention rate of 0.77% per pharmacy (SD = 0.61%). There was a large variability among pharmacies (0-2.6%). Most of the clinical DRPs were associated with dosage problems (n = 91) and drug-drug interactions (n = 45). The most frequent potential negative outcomes reported were quantitative inefficacy (n = 101) and quantitative safety (n = 94). Two-thirds of clinical DRPs required a prescription modification (n = 186), the most frequent being a change in dosage or drug regimen. In 110 interventions (38%), physicians were immediately contacted to take part in the decision. In 122 interventions (43%), pharmacists managed the interventions alone. However, in 55 interventions (19%), pharmacists managed the DRPs with the patient. From these 287 clinical interventions, 134 different codes were reported. Seven hundred and thirty-six technical problems related to prescriptions required intervention, which corresponded to a mean intervention rate of 1.90% per pharmacy. The main type of problem was a discrepancy with the medication record (n = 208). There were 494 instances that required a prescription modification. Pharmacists resolved 45% of all technical problems by themselves. CONCLUSION: The developed coding system could describe the management process for DRPs. The observed intervention rate and the frequency of steps involved were comparable to those previously observed for pharmacists' interventions. Data regarding the entire process used to manage drug-related problems can be useful in improving medication safety, education, and collaborative care. PMID- 18392733 TI - [Tests for permanent reform of long term care insurance]. PMID- 18392734 TI - [Case management in long-term care--the task of individual related and family oriented support for people in need of care and its realization through the reform of long-term care insurance]. AB - The introduction of standardized Case Management structures to improve coordination and cooperation of all involved in care, such as cost units, service providers, voluntary organizations, families and the different occupational categories involved in nursing, is the main concern of the current reform of German long-term care insurance. In this article, demands on Case Management in care are enunciated and the basics found in expert talks, needed for efficient support of care, assembled. In doing so, the role and function of Case Management is differentiated, the different levels (case, organizational and system levels) distinguished and options and conditions needed to settle such an organization are introduced. PMID- 18392735 TI - [Community Care Access Centers--a way to further develop local care policy in Germany]. AB - The recent reform act on long-term care insurance includes plans for the establishment of Community Care Access Centers. This paper addresses chances and challenges of these centers and proposes some important steps for program development. PMID- 18392737 TI - Lignan glycosides from the leaves of Osmanthus heterophyllus. AB - Seven known lignan glycosides were isolated from the leaves of Osmanthus heterophyllus: (+)-syringaresinol 4-O-beta-D-glucopyranoside, (+)-syringaresinol 4, 4'-O-di-beta-D-glucopyranoside, (+)-medioresinol 4, 4'-O-di-beta-D glucopyranoside, (+)-medioresinol 4-O-beta-D-glucopyranoside, (+)-pinoresinol 4, 4'-O-beta-D-glucopyranoside, (+)-epipinoresinol 4-O-beta-D-glucopyranoside and phillyrin. Their structures were determined on the basis of spectral data. PMID- 18392736 TI - Idiopathic collapsing glomerulopathy in children. AB - BACKGROUND: Collapsing glomerulopathy (CG) is a clinically and pathologically distinct variant of focal segmental glomerulosclerosis (FSGS). Pathologically similar lesions have been reported in adults and children with human immuno deficiency virus (HIV) infection. However, there is a recent interest in the recognition of this variant in the absence of HIV infection. AIM: To evaluate the clinical presentation and outcome of our pediatric patients with idiopathic CG. METHODS: A sum of six children with idiopathic CG, aged 1-7 years at presentation, were retrospectively identified. Clinical data and renal biopsy were reviewed for all patients. Serum creatinine and estimated GFRs at presentation and last follow-up were compared using the Wilcoxon signed rank test and the risk factors for occurrence of ESRD analyzed using the Cox proportional hazard models. RESULTS: Steroid-resistant nephrotic syndrome with or without azotemia was the presenting clinical finding in all the cases. The median serum creatinine values at onset and last follow-up were 1.05 and 1.25 mg/dl, respectively (p = 0.128). Following immunosuppressive therapy one patient achieved complete remission of proteinuria, and four were in partial remission. The remaining one patient did not show any change in proteinuria at 6 months of therapy. Two of the six patients progressed to end-stage renal disease within a median follow-up period of 27 months (range 14-96 months). CONCLUSIONS: Collapsing glomerulopathy is an aggressive variant of focal segmental glomerulosclerosis. All patients with CG should be screened for the underlying etiology, and patients with idiopathic CG should be offered a trial of immunosuppressive therapy. PMID- 18392738 TI - Triterpenes acids and saponins isolated from Licania arianeae Prance (Chrysobalanaceae). AB - From the leaves and wood of Licania arianeae, ten known compounds were isolated and identified. They belong to pharmacological active triterpenes acids, including three oleanoic acids, five ursane acids, and two triterpenes saponin acids. Their structures were established by analysis of infrared, mass spectrometry, and nuclear magnetic resonance spectral data of natural triterpenes and of methyl acetyl derivatives. PMID- 18392739 TI - Congenital disorders of glycosylation--a challenging group of IEMs. AB - Congenital disorders of glycosylation (CDG) are a rapidly growing group of inherited errors of metabolism (IEMs) due to an impairment of one or several glycosylation pathways. During recent years over 30 CDG subtypes have been identified at a molecular and biochemical level. The clinical manifestations in CDG are heterogeneous and may be highly variable within the same subtype and even among affected siblings. Novel insights into the extremely complex glycosylation pathways have necessitated several reclassifications of the group of CDG. Today CDG comprise not only the formerly known multisystem glycosylation defects but also some tissue-specific glycosylation defects, implicating a different diagnostic work-up depending on the underlying glycosylation defect. In 2007 the expanding group of CDG is an enormous challenge to all specialists working in the field of IEMs. This review gives a brief overview about the expanded group of CDG and summarizes the main implications for clinicians. PMID- 18392740 TI - PPARs as therapeutic targets for correction of inborn mitochondrial fatty acid oxidation disorders. AB - Enzyme defects in the mitochondrial fatty acid oxidation (FAO) are a large family of inherited metabolic disease well characterized clinically and genetically, but for which pharmacological strategies remain limited. It is now well established that regulation of genes involved in mitochondrial FAO is under control of the PPAR (peroxisome proliferator activated receptor) signalling pathway, and this led us to test a possible pharmacological correction of FAO disorders by fibrates and other PPAR activators. This review presents the basic data supporting our initial hypothesis, summarizes the results obtained in cells from patients with CPT II (carnitine palmitoyltransferase II) or VLCAD (very long-chain acyl-CoA dehydrogenase) deficiency, and discusses the perspectives and limits of this approach for therapy of these disorders. PMID- 18392741 TI - Valproic acid metabolism and its effects on mitochondrial fatty acid oxidation: a review. AB - Valproic acid (VPA; 2-n-propylpentanoic acid) is widely used as a major drug in the treatment of epilepsy and in the control of several types of seizures. Being a simple fatty acid, VPA is a substrate for the fatty acid beta-oxidation (FAO) pathway, which takes place primarily in mitochondria. The toxicity of valproate has long been considered to be due primarily to its interference with mitochondrial beta-oxidation. The metabolism of the drug, its effects on enzymes of FAO and their cofactors such as CoA and/or carnitine will be reviewed. The cumulative consequences of VPA therapy in inborn errors of metabolism (IEMs) and the importance of recognizing an underlying IEM in cases of VPA-induced steatosis and acute liver toxicity are two different concepts that will be emphasized. PMID- 18392743 TI - Reduction in bone mineral density in glycogenosis type III may be due to a mixed muscle and bone deficit. AB - Glycogen storage disease type III (GSD III; OMIM 232400) is an autosomal recessive deficiency of the glycogen debrancher enzyme, amylo-1,6-glucosidase (EC 3.2.1.33). Patients with other hepatic glycogenoses are known to have reduced bone mineral content (BMC) and to be at consequent risk of fractures. They have key metabolic differences from GSD III patients, however. This study examines bone density and metabolism in 15 GSD III patients (6 female) from childhood to adulthood (aged 10-34 years). The results demonstrate that patients with GSD III have low bone mass at all skeletal sites compared with healthy individuals of the same age and sex, with a significant proportion (40-64%) having BMD > 2 standard deviations below the mean for whole body and lumbar spine. The deficiency seems to be attributable to a mixed muscle andbone deficit. Lower bone mass was found at all sites for GSD IIIa patients (combined liver and muscle defect) compared with GSD IIIb patients (liver only defect). CONCLUSION: Patients with GSD III have significantly abnormal bone mass, placing them at increased risk of potential fracture. The underlying mechanism is probably multifactorial with contributions from abnormal muscle physiology, abnormal metabolic milieu and altered nutrition affecting micronutrient intake. Therapies need to address all these factors to be successful. PMID- 18392744 TI - Liver cell transplantation for the treatment of inborn errors of metabolism. AB - Over the last 15 years, liver cell transplantation (LCT) has developed from an experimental laboratory technique to a potentially life-saving therapeutic option. Because of its minimally invasive nature, the method is especially attractive for (small) children. In children with liver-based inborn errors of metabolism, this transfer of enzyme activity can be regarded as a gene therapy, which can be installed independently and additionally to conservative treatment concepts. To date 14 children with inherited metabolic diseases have undergone LCT in various centres. Although individual results are encouraging, different treatment protocols, difficulties in the objective assessment of function of the transplant, and finally the lack of a controlled study make it difficult to judge the overall significance of LCT in the treatment of metabolic diseases and call for collaborative clinical research. PMID- 18392742 TI - Sanfilippo syndrome: a mini-review. AB - Mucopolysaccharidosis type III (MPS III, Sanfilippo syndrome) is an autosomal recessive disorder, caused by a deficiency in one of the four enzymes involved in the lysosomal degradation of the glycosaminoglycan heparan sulfate. Based on the enzyme deficiency, four different subtypes, MPS IIIA, B, C, and D, are recognized. The genes encoding these four enzymes have been characterized and various mutations have been reported. The probable diagnosis of all MPS III subtypes is based on increased concentration of heparan sulfate in the urine. Enzymatic assays in leukocytes and/or fibroblasts confirm the diagnosis and allow for discrimination between the different subtypes of the disease. The clinical course of MPS III can be divided into three phases. In the first phase, which usually starts between 1 and 4 years of age, a developmental delay becomes apparent after an initial normal development during the first 1-2 years of life. The second phase generally starts around 3-4 years and is characterized by severe behavioural problems and progressive mental deterioration ultimately leading to severe dementia. In the third and final stage, behavioural problems slowly disappear, but motor retardation with swallowing difficulties and spasticity emerge. Patients usually die at the end of the second or beginning of the third decade of life, although survival into the fourth decade has been reported. Although currently no effective therapy is yet available for MPS III, several promising developments raise hope that therapeutic interventions, halting the devastating mental and behavioural deterioration, might be feasible in the near future. PMID- 18392745 TI - Disorders caused by deficiency of succinate-CoA ligase. AB - Succinate-CoA ligase catalyses the reversible conversion of succinyl-CoA and ADP or GDP to succinate and ATP or GTP. It is a mitochondrial matrix enzyme and at least the ADP-forming enzyme is part of the Krebs cycle. The substrate specificity is determined by the beta subunit of succinate-CoA ligase, which is encoded by either SUCLA2 or SUCLG2. In patients with severe hypotonia, deafness and Leigh-like syndrome, mutations have been found in SUCLA2. Mutations have also been reported in SUCLG1, which encodes the alpha subunit found in both enzymes, in patients with severe infantile acidosis and lactic aciduria. Elevated methylmalonate and methylcitrate and severe mtDNA depletion were found in both disorders. The mtDNA depletion may be explained by the interaction of succinate CoA ligase with nucleoside diphosphate kinase, which is involved in mitochondrial nucleotide metabolism. PMID- 18392746 TI - AGAT, GAMT and SLC6A8 distribution in the central nervous system, in relation to creatine deficiency syndromes: a review. AB - Creatine deficiency syndromes, either due to AGAT, GAMT or SLC6A8 deficiencies, lead to a complete absence, or a very strong decrease, of creatine within the brain, as measured by magnetic resonance spectroscopy. While the mammalian central nervous system (CNS) expresses AGAT, GAMT and SLC6A8, the lack of SLC6A8 in astrocytes around the blood-brain barrier limits the brain capacity to import creatine from the periphery, and suggests that the CNS has to rely mainly on endogenous creatine synthesis through AGAT and GAMT expression. This seems contradictory with SLC6A8 deficiency, which, despite AGAT and GAMT expression, also leads to creatine deficiency in the CNS. We present novel data showing that in cortical grey matter, AGAT and GAMT are expressed in a dissociated way: e.g. only a few cells co-express both genes. This suggests that to allow synthesis of creatine within the CNS, at least for a significant part of it, guanidinoacetate must be transported from AGAT- to GAMT-expressing cells, possibly through SLC6A8. This would explain the creatine deficiency observed in SLC6A8-deficient patients. By bringing together creatine deficiency syndromes, AGAT, GAMT and SLC6A8 distribution in CNS, as well as a synthetic view on creatine and guanidinoacetate levels in the brain, this review presents a comprehensive framework, including new hypotheses, on brain creatine metabolism and transport, both in normal conditions and in case of creatine deficiency. PMID- 18392747 TI - Functional antibody deficiency in a patient with type I Gaucher disease. AB - Gaucher disease (GD), the most common lysosomal storage disorder, demonstrates an autosomal recessive pattern of inheritance. The genetic defect in GD leads to decreased production of the lysosomal enzyme glucosylceramide hydrolase, thereby resulting in the deposition of glucosylceramide sphingolipids within multiple organ systems. Although the precise mechanisms remain unclear, GD is usually associated with chronic antigenic stimulation and hyperimmunoglobulinaemia. We report a novel case of type I GD coexisting with relatively low serum immunoglobulins, impaired antibody production, and recurrent bacterial infections in a 62-year-old male. The patient had been diagnosed with GD 30 years previously and had subsequently started enzyme replacement therapy. Since being diagnosed with GD, the patient had suffered from repeated episodes of acute bronchitis and a recent severe bout of community-acquired pneumonia that required a lengthy hospitalization. On our initial evaluation, the patient had laboratory testing that demonstrated: decreased serum IgG, IgG2, and IgA levels; reduced absolute CD3(+)/CD4(+), CD3(+)/CD8(+), and lymphocyte counts; low IgG titres to pneumococcal polysaccharide vaccine; and decreased anti-tetanus antibodies. Lymphocyte function analysis demonstrated a normal response to phytohaemagglutinin, and decreased responses to concanavalin A and pokeweed mitogen. Repeat testing after 6 months revealed normal serum immunoglobulin levels and mitogenic responses. Although the explanation for our observed transient hypogammaglobulinaemia remains unclear, this patient's clinical constellation (i.e. repeated infections, hypogammaglobulinaemia and lymphopenia, decreased post-vaccination titres, and impaired responses to some mitogens) shares overlapping features with common variable immunodeficiency (CVID). PMID- 18392749 TI - Multisystem involvement in a patient due to accumulation of amylopectin-like material with diminished branching enzyme activity. AB - We report a 13-year-old boy with multisystem involvement secondary to accumulation of amylopectin-like material. He was born to consanguineous parents at full term without any complications and his maternal perinatal history was uneventful. His parents were cousins. He had normal growth and development except for his weight. His sister died from an unexplained cardiomyopathy at the age of 8 years. Our patient's initial symptom was severe heart failure. Since he also had a complaint of muscle weakness, electromyography was performed which showed muscle involvement. The diagnosis was suggested by tissue biopsy of skeletal muscle showing intracellular, basophilic, diastase-resistant, periodic acid Schiff-positive inclusion bodies and was confirmed by the presence of a completed branching enzyme deficiency. Similar intracytoplasmic inclusion-like bodies were also found in liver biopsy, but very few in number compared with the skeletal muscle. The patient died from an intercurrent infection. Postmortem endomyocardial biopsy revealed the same intracytoplasmic inclusions as described above affecting almost all myocardial cells. Ultrastructural examination of liver biopsy was nondiagnostic; however, myocardium showed prominent, large, intracytoplasmic deposits. Glycogen branching enzyme gene sequence was normal, and thus classical branching enzyme deficiency was excluded. Our patient represents the first molecular study performed on a patient in whom there was multiple system involvement secondary to accumulation of amylopectin-like material. We suggest that this is an as yet undefined and different phenotype of glycogen storage disease associated with multisystemic involvement. PMID- 18392748 TI - Pathogenesis of CNS involvement in disorders of amino and organic acid metabolism. AB - Inherited disorders of amino and organic acid metabolism have a high cumulative frequency, and despite heterogeneous aetiology and varying clinical presentation, the manifestation of neurological disease is common. It has been demonstrated for some of these diseases that accumulating pathological metabolites are directly involved in the manifestation of neurological disease. Various pathomechanisms have been suggested in different in vitro and in vivo models including an impairment of brain energy metabolism, an imbalance of excitatory and inhibitory neurotransmission, altered transport across the blood-brain barrier and between glial cells and neurons, impairment of myelination and disturbed neuronal efflux of metabolic water. This review summarizes recent knowledge on pathomechanisms involved in phenylketonuria, glutaric aciduria type I, succinic semialdehyde dehydrogenase deficiency and aspartoacylase deficiency with examples, highlighting general as well as disease-specific concepts and their putative impact on treatment. PMID- 18392750 TI - Hepatic cirrhosis, dystonia, polycythaemia and hypermanganesaemia--a new metabolic disorder. AB - We report a new constellation of clinical features consisting of hypermanganesaemia, liver cirrhosis, an extrapyramidal motor disorder and polycythaemia in a 12 year-old girl born to consanguineous parents. Blood manganese levels were >3000 nmol/L (normal range <320 nmol/L) and MRI revealed signal abnormalities of the basal ganglia consistent with manganese deposition. An older brother with the same phenotype died at 18 years, suggesting a potentially lethal, autosomal recessive disease. This disorder is probably caused by a defect of manganese metabolism with the accumulation of manganese in the liver and the basal ganglia similar to the copper accumulation in Wilson disease. In order to assess the genetic basis of this syndrome we investigated two candidate genes: ATP2C2 and ATP2A3 encoding the manganese-transporting calcium ATPases, SPCA2 and SERCA3, respectively. Genotyping of the patient and the family for microsatellite markers surrounding ATP2C2 and ATP2A3 excluded these genes. The patient was found to be heterozygous for both gene loci. Despite the unknown pathophysiology, we were able to develop a successful treatment regime. Chelation therapy with disodium calcium edetate combined with iron supplementation is the treatment of choice, lowering blood manganese levels significantly and improving clinical symptoms. PMID- 18392751 TI - The relation of cerebrospinal fluid and plasma glycine levels in propionic acidaemia, a 'ketotic hyperglycinaemia'. AB - The characteristic elevation of plasma glycine concentrations observed in propionic acidaemia (PA) and other 'ketotic hyperglycinaemias' has been attributed to secondary inhibition of the hepatic glycine cleavage system (GCS) by accumulating CoA derivatives of branched-chain amino acid metabolites. In nonketotic hyperglycinaemia (NKH), cerebrospinal fluid (CSF) and plasma glycine levels and their ratio are increased due to primary deficiency of central nervous system (CNS) as well as hepatic GCS. Whether the GCS in the CNS is also inhibited in PA is unclear, as there are scant data available on CSF glycine levels in this disorder. We studied the relation of CSF and plasma glycine levels in 6 paired samples from 4 PA patients, including one PA patient with bacterial meningitis who underwent ventriculoperitoneal shunting and multiple CSF analyses (n = 26). In contrast to the CSF glycine levels which were generally elevated in all four PA patients, the CSF/plasma glycine concentration ratios in paired samples were normal (0.016-0.029), with the exception of a single sample (0.132) with extremely high CSF protein concentration (2010 mg/L) during the course of meningitis indicating a disturbed blood-brain barrier. This finding of normal CSF/plasma glycine ratio in PA suggests that the observed elevations of CSF glycine levels are a reflection of the concurrent hyperglycinaemia resulting from secondary inhibition of hepatic GCS, but that brain GCS is not affected, in contrast to the situation in NKH. The neurological sequelae in PA are therefore unlikely to be related to disturbed glycine metabolism. PMID- 18392752 TI - Glucose-6-phosphate dehydrogenase deficiency protects against coronary heart disease. AB - Previous studies suggest a reduction in cardiovascular risk among subjects expressing the glucose-6-phosphate dehydrogenase (G6PD, EC 1.1.1.49) deficient phenotype. We aimed to test this hypothesis in male subjects expressing the G6PD deficient phenotype vs wild type G6PD. In a case-control study we examined consecutive patients admitted for acute myocardial infarction or unstable angina, and controls admitted for diagnoses other than coronary heart disease (CHD). The G6PD phenotype was determined by measuring the enzyme activity in erythrocytes, as the absorbance rate change due to NADPH reduction. The CHD risk associated with the G6PD phenotype was assessed with unconditional logistic regression. G6PD deficient subjects were less frequently represented among cases (11.8%) than among controls (18.6%, p=0.002). The genetic condition of G6PD deficiency conveyed a significant reduction in CHD risk (OR=0.6; 95% CI 0.4 to 0.9). We confirm the hypothesis that subjects with the G6PD-deficient phenotype are less prone to CHD. We suggest that such a protective effect may be ascribable to a reduced 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA R) activity, a statin-like effect, as well as to a downregulation in NADPH oxidase activity with a consequent reduction in oxygen-free radical production. PMID- 18392753 TI - Comparison of haloperidol, risperidone, sertindole, and modafinil to reverse an attentional set-shifting impairment following subchronic PCP administration in the rat--a back translational study. AB - RATIONALE: Selective cognitive impairments, including those of executive function as assessed using the Wisconsin Card Sort Test or intradimensional extradimensional (ID-ED) tests, are a key feature of schizophrenia but remain inadequately treated by existing therapies. Recently, however, modafinil has been shown to improve attentional set-shifting performance in patients with schizophrenia. OBJECTIVE: The present study evaluated the recently described analogous rat ID-ED attentional set-shifting task by investigating the effects of various pharmacological challenges to a phencyclidine (PCP)-induced ED shift impairment, namely, haloperidol, risperidone, sertindole, and modafinil. MATERIALS AND METHODS: Rats were subjected to a subchronic systemic administration of either saline vehicle or PCP (5 mg/kg i.p. b.i.d. for 7 days) followed by a 7-day washout period. During this period, rats were trained to dig in baited bowls for a food reward and to discriminate based on odor or digging media. In a single test session conducted the day after the washout period (day 8), rats performed a series of discriminations following acute administration of either vehicle, or haloperidol (0.1 mg/kg s.c.), or risperidone (0.2 mg/kg i.p.), or sertindole (1.25 mg/kg p.o.) or modafinil (64 mg/kg p.o.). RESULTS: The subchronic PCP-induced ED deficit was ameliorated by sertindole and modafinil but not by haloperidol or risperidone. CONCLUSIONS: Overall, these findings further support that the rat ID-ED test in subchronic PCP-treated rats has utility and validity as a preclinical model of the cognitive symptoms of schizophrenia and demonstrates back-translational potential. PMID- 18392754 TI - Increased efficacy of micro-opioid agonist-induced antinociception by metabotropic glutamate receptor antagonists in C57BL/6 mice: comparison with (-) 6-phosphonomethyl-deca-hydroisoquinoline-3-carboxylic acid (LY235959). AB - RATIONALE: Recent experimental data suggest that metabotropic glutamate receptor (mGluR) antagonists with selectivity for mGluR1 and mGluR2/3 enhance morphine induced antinociception. OBJECTIVES: The present study addressed the hypothesis that mGluR antagonists enhance opioid antinociception by increasing opioid efficacy. MATERIALS AND METHODS: The antinociceptive effects of the partial mu opioid receptor agonists buprenorphine and dezocine were first assessed in a hot plate procedure under conditions of low (53 degrees C) and high (56 degrees C) stimulus intensity. Under conditions in which buprenorphine and dezocine produced submaximal antinociceptive effects, these drugs were assessed after pretreatment with the mGluR1 antagonist JNJ16259685, the mGluR5 antagonist MPEP, the mGluR2/3 antagonist LY341495, and for comparison, the N-methyl-D-aspartate (NMDA) receptor antagonist LY235959. RESULTS: Buprenorphine (0.032-3.2 mg/kg) and dezocine (0.1 10 mg/kg) were fully efficacious at 53 degrees C and produced submaximal antinociceptive effects at 56 degrees C (i.e., their effects did not exceed 50% of the maximum possible effect). Pretreatment with JNJ16259685 (1.0-3.2 mg/kg), LY341495 (1.0-3.2 mg/kg), and LY235959 (0.32-1.0 mg/kg) enhanced the antinociceptive effects of buprenorphine and dezocine at 56 degrees C, as revealed by significant increases in the peak effects of both drugs to approximately 100% maximum possible effect. In contrast, pretreatment with MPEP (1.0-3.2 mg/kg) did not modulate the antinociceptive effects of buprenorphine and dezocine. CONCLUSIONS: These results suggest that, similar to the NMDA receptor antagonist LY235959, the mGluR1 antagonist JNJ16259685 and the mGluR2/3 antagonist LY341495 increase the antinociceptive efficacy of buprenorphine and dezocine. PMID- 18392755 TI - Restricted-access media development for direct analysis of drugs in biofluids using capillary liquid chromatography. AB - In analytical sciences the design of novel materials and stationary phases for the sample preparation and separation of analytes from biological fluids is needed. In this work we present different strategies for modification of stationary phases to produce tailored solutions for the analytical problem. In this context a novel shielded polymeric reversed-phase monolithic material was prepared in the presence of different numbers of reactive groups and concentrations of the coating polymer. Chromatographic experiments were performed using benzoic acid propyl ester in order to characterize the hydrophobicity and efficiency of the different restricted-access continuous beds prepared. Inverse size-exclusion chromatography was used for investigation of the pore structure properties of the beds. Capillary columns were applied for nanochromatography of biological fluids containing a mixture of nitrazepamum and medazepamum. PMID- 18392759 TI - Phylogenetic and structural analysis of translationally controlled tumor proteins. AB - The translationally controlled tumor protein (TCTP) is conserved in all eukaryotes studied thus far. Recent evidence points to an important role for TCTP in the induction of cell proliferation in animals through an interaction with G proteins. TCTP may also constitute an intercellular secreted signal that modulates the immune response in the vertebrates. Because of its sequence conservation and ubiquity, the analysis of its amino acid sequence divergence between different taxa may provide insight into the structural constraints on the evolution of this protein. In the present study, we analyzed the phylogeny of TCTP sequences from a wide range of organisms and found that, with some exceptions, the groupings formed were consistent with the evolutionary history. Indeed, at the level of lower-order taxa, the groupings are in agreement with their established phylogeny, thus indicating that the substitution rates of the TCTP residues varied evenly between members of the same clade. Predicted three dimensional structures of representative TCTPs, based on the reported 3D structure of Schizosaccharomyces pombe, indicated that these proteins are highly conserved among diverse taxonomic groups. However, analysis of the primary structure indicated subtle differences in the domain-forming pocket that potentially interacts with G proteins, particularly among Diplomonadidae, Apicomplexa, and other parasites of vertebrates. These differences support the notion that these specific TCTPs could block the normal immune response by acting as dominant negative mutants. Structural differences were also observed in a reported sequence of TCTP from Plasmodium knowlesi, in which the presence of an extra alpha-helix could also interfere in the interaction with G proteins. PMID- 18392760 TI - Expression pattern diversity and functional conservation between retroposed PRAT genes from Drosophila melanogaster and Drosophila virilis. AB - Gene duplication by retrotransposition duplicates only the coding and untranslated regions of a gene and, thus, biases retroduplicated genes toward having different expression patterns from their parental genes. As such, genes duplicated by retrotransposition are more likely to develop novel expression domains. To explore this idea further, we used the Prat/Prat2 gene duplication in Drosophila as a case study to examine the aftermath of a retrotransposition event that resulted in both the parent and the child gene becoming essential for survival. We used the Gal4-UAS transgene system with EGFP as a reporter to determine the developmental expression patterns of Prat and Prat2 from D. melanogaster (DmPrat and DmPrat2) and Prat from D. virilis (DvPrat). We also tested the functional equivalence of the protein products of DmPrat and DmPrat2. We found that each of the proteins could rescue DmPrat mutations, showing that the requirement for both Prat and Prat2 in Drosophila is not simply due to differences in protein function. In contrast, we found that the DmPrat and DmPrat2 genes have developed nonoverlapping patterns of expression, which correlate with their respective loss-of-function phenotypes. We further found that DvPrat expression is similar to DmPrat during development but differs in adult gonads. Thus, the function of the Prat retrogene has not diverged in the D. melanogaster and D. virilis lineages, while some aspects of its expression pattern have evolved. Finally, we have identified promoter elements, conserved upstream of DmPrat and DvPrat, that this retrogene has acquired to drive its expression. PMID- 18392762 TI - Type II opsins: evolutionary origin by internal domain duplication? AB - Opsins are a large group of proteins with seven transmembrane segments (TMSs) that are found in all domains of life. There are two types of opsins that are sometimes considered nonhomologous: type I is known from prokaryotes and some eukaryotes, while type II is known only from Eumetazoan animals. Type II opsins are members of the family of G-protein coupled receptors (GPCRs), which facilitate signal transduction across cell membranes. While previous studies have concluded that multiple transmembrane-containing protein families-including type I opsins-originated by internal domain duplication, the origin of type II opsins has been speculated on but never tested. Here we show that type II opsins do not appear to have originated through a similar internal domain duplication event. This provides further evidence that the two types of opsins are nonhomologous, indicating a convergent evolutionary origin, in which both groups of opsins evolved a seven-TM structure and light sensitivity independently. This convergence may indicate an important role for seven-TM protein structure for retinal-based light sensitivity. PMID- 18392761 TI - Selection on length mutations after frameshift can explain the origin and retention of the AP3/DEF-like paralogues in Impatiens. AB - Evolution of class B genes through gene duplication has been proposed as an evolutionary mechanism that contributed to the enormous floral diversity. Frameshift mutations are a likely mechanism to explain the divergent C-terminal sequences of MIKC gene subfamilies. So far, the inferences for frameshifts and selective pressures on the C-terminal domain are made for old duplications for which the exact selective pressures are obscured by evolutionary time. This motivated us to study an example of a recent duplication, which allows us to consider in more detail the selective pressures that are involved after duplication. We find that after duplication and frameshift of Impatiens class B genes, the individual codons show no evidence for adaptive selection. It is rather the length of the C-terminal domain that either is strictly conserved or varies strongly. This suggests a role for the length of the C-terminal domain in the retention of duplicated genes. PMID- 18392763 TI - Metrical and non-metrical study of anterior clinoid process in South Indian adult skulls. AB - BACKGROUND: The complex architecture of the anterior clinoid process (ACP), which is usually removed during the surgical elimination of tumors or aneurysms of sellar region, has surgical importance. For effective clinoidectomy, a neurosurgeon must have the prior knowledge of anatmoical variations of ACP. The purpose of this study was to investigate the dimensions and variation in the shape of ACP in dry adult skulls of South Indian origin. METHODS: The study was conducted on 25 dry adult South Indian skulls of either sex. Basal width, length and thickness of ACP were measured on both the sides using Vernier caliper. Non metrical parameters such as shape, direction of ACP were recorded. RESULTS: ACP exhibited different anatomical variations with respect to their shape and direction. Triangular, pentagonal, nipple shaped, J-shaped and finger like ACP's were observed in these skulls. Bilaterally triangular ACP was the most common type observed (64%). ACP was bilaterally straight in 68% of the skulls and bilaterally curved in 16% of the skulls. ACP with blunt end and pointed end was observed bilaterally in 52 and 24% of the skulls, respectively. The average length, basal width and thickness of ACP on right side was 10.68 +/- 1.90, 12.4 +/- 2.58 and 6.88 +/- 1.09 mm, respectively, and on the left side it was 9.96 +/- 1.71, 11.12 +/- 1.81 and 6.52 +/- 0.96 mm, respectively. CONCLUSIONS: The result of the present investigation suggests that ACPs of South Indian skulls are highly variable and, are marginally larger and thicker than ACPs of Nepalese and Korean origin. PMID- 18392764 TI - Dural arachnoid granulations and "giant" arachnoid granulations. AB - Although arachnoid granulations (AGs) were already described by Antonio Pacchioni more than 300 years ago, two issues draw particular attention: first, the radiological features and differential diagnosis of the so-called giant AGs (GAGs) and second, their possible association with various disease processes. In order to evaluate the frequency, size and normal distribution of GAGs, an anatomical study of the dural sinuses was carried out. It involved all the autopsies performed during the period August 2002-February 2005 and included 651 cases: 306 females and 345 males, aged 13-99 years (mean 69 years). Grossly visible GAGs were identified in 24 cases: 7 females and 17 males, aged 45-92 years (mean 69 years). This is the largest population-based anatomical study on GAGs. It shows that GAGs, in general a rare finding (3.68%), are rather common in the adult population, especially in the elderly (aged >65 years) and that they can reach remarkable size (up to 2.5 cm and more in diameter). Giant AGs should be considered in the radiological differential diagnosis of intradural lesions, particularly those occurring in the transverse sinus of the elderly. PMID- 18392765 TI - Unilateral arteria peronea magna associated with bilateral replaced dorsalis pedis arteries. AB - During anatomic dissection, a 59-year-old man presented with an arteria peronea magna that replaced all the branches of the posterior tibial artery as well as the dorsalis pedis artery. Thus, the right popliteal artery terminated into the anterior tibial artery and the enlarged peroneal artery. The right anterior tibial artery ended before it reached the ankle. The right dorsalis pedis artery arose from the peroneal artery through its perforating branch. The left popliteal artery gave rise to a common stem that subdivided into two individual branches replacing the proximal part of the anterior tibial artery, and then divided into the posterior tibial artery and the peroneal artery. As on the right side, the left dorsalis pedis artery also arose from the peroneal artery through its perforating branch. A possible embryologic explanation of these variations is proposed. PMID- 18392766 TI - Platelets in regenerative medicine. AB - Stem and progenitor cells have evolved as the central cell type in regenerative medicine. This focussed approach may, however, sometimes narrow the point of view. Recently, platelets have been shown to strongly influence progenitor cell biology and to serve as regenerative cells themselves. Platelets represent an important recruitment factor for circulating progenitor cells. Furthermore, they modulate progenitor cell chemotaxis, migration and differentiation. Platelets are crucial for the regeneration of parenchymatous organs such as the liver. Moreover, in the clinical setting, platelets are already applied for the restoration of connective and bone tissue. However, also adverse effects can be mediated by platelets and progenitor cells exposed to platelets, including the induction of pro-inflammatory processes. This review gives an update on platelets as regenerative cells and as modulators of progenitor cell biology in (patho-) physiological conditions. Abundant data obtained during the last decade have linked progenitor and stem cells to tissue repair. Very recent accumulating results from different groups revealed that platelets may have a strong influence on central functions of these progenitor cells. Only little is known about the impact of platelets themselves on tissue regeneration. Therefore, this review highlights the role of platelets themselves as a potential effector cell for tissue regeneration and their impact on progenitor cell biology. PMID- 18392767 TI - Regional and temporal heterogeneity of postsystolic wall thickening is associated with left ventricular asynchrony in normal and experimental stunned myocardium. AB - AIMS: Postsystolic wall thickening (PSWT) occurs after aortic valve closure. We investigated the influence of ischemia location and wall interactions on PSWT in normal and stunned myocardium. METHODS AND RESULTS: Twenty-two dogs were studied. Seven chronically instrumented dogs (sonomicrometry) underwent 10-min coronary artery occlusion (CAO) of left circumflex artery ("LCX stunning") and seven other dogs underwent 10-min CAO of the anterior descending artery ("LAD stunning") followed by reperfusion. At baseline, there was no PSWT in the anterior wall whereas posterior wall started and finished to thicken after the anterior wall, demonstrating PSWT. With LCX stunning, PSWT was observed in the posterior wall without affecting the remote anterior wall. With LAD stunning, PSWT in the anterior wall was transient and of lower magnitude Vs. posterior wall; in the remote posterior wall, PSWT previously observed at baseline, almost vanished. Postsystolic to systolic wall thickening ratio identified (ROC analysis) normal, ischemic and stunned myocardium with different amplitudes between walls. Tissue Doppler Imaging demonstrated similar pattern in basal, mid and apical segments (additional n = 4 for both LCX and LAD stunning). CONCLUSION: The present study demonstrates that location of ischemia and wall interactions produce discrepancies in PSWT between anterior and posterior walls in stunned myocardium. PMID- 18392768 TI - Genes in congenital heart disease: atrioventricular valve formation. AB - Through the use of animal studies, many candidate genes (mainly encoding transcriptional factors and receptors) have been implicated in the development of congenital heart disease. Thus far, only a minority of these genes have been shown to carry mutations associated with congenital disease in humans, e.g., GATA 4, TBX-5, NOTCH1 and NKX2-5. Mutations in these genes can cause a variety of cardiac defects even within the same family. Conversely, similar phenotypes are observed for different gene mutations suggesting a common pathway. Multiple genes and genetic pathways have been related to atrioventricular valve formation, although most of these genes have not yet been demonstrated as causative in human atrioventricular valve defects. Key pathways include the epidermal growth factor receptor pathway and related interacting pathways, most importantly the pathway of UDP-glucose dehydrogenase, resulting ultimately in activation of Ras. Other examples of interacting pathways include that of Nodal/Cited2/Pitx2, Wnt, Notch and ECE. Further studies are needed to investigate the pathways which are crucial for atrioventricular valve formation in humans. Understanding the underlying molecular process of abnormal atrioventricular valve formation in patients with congenital heart disease may provide important insight, in the etiology and possibly into preventive or treatment regimes. PMID- 18392769 TI - Efficient rescue of infectious bursal disease virus using a simplified RNA polymerase II-based reverse genetics strategy. AB - We describe a simplified RNA polymerase II-based reverse genetics approach that allows for the efficient rescue of high-titer infectious bursal disease virus (IBDV) from cloned cDNAs of genomic segments A and B. Unlike the previously reported RNA polymerase II-based methods, the developed strategy does not necessitate the introduction of a ribozyme sequence at both ends of the genomic cDNA sequences. This was achieved by fusing the 5' terminal sequence of the cDNA of each segment to the transcription start site of the immediate early cytomegalovirus promoter, while a ribozyme sequence was only introduced at the 3' end. Using this strategy, and without complementing with IBDV structural proteins, titers as high as 10(11) tissue culture infectious dose 50 were reproducibly obtained in chicken embryo fibroblast cells immediately upon co transfection with cDNAs of both segments. We anticipate that this modification could improve reverse genetics for any other RNA virus and may be beneficial for vaccine development and dissection of the viral life cycle. PMID- 18392770 TI - Molecular detection and genetic diversity of norovirus in hospitalized young adults with acute gastroenteritis in Bahia, Brazil. AB - The molecular epidemiology of a recent norovirus (NoV) outbreak in Brazil performed by comparative analysis with Genebank NoV sequences showed that the GII.4 strain was responsible for 72.5% of all NoV-positive cases (58/80). Other detected NoV strains included GII.3 (7/80; 8.8%) and GII.9 (8/80; 10%). This is the first outbreak reported in Bahia state, Brazil, during June-July of 2006, where NoV was identified as the principal etiologic agent in hospitalized young adults with acute gastroenteritis symptoms. These findings suggest that GII.4 is a predominant circulating genotype in NoV outbreaks in Brazil. PMID- 18392771 TI - Hypotransferrinemia and changes in plasma lipid and metabolic patterns in sepsis. AB - This study was performed to obtain a characterization of the changes in plasma transferrin (Tf, g/L) in sepsis. More than four hundred determinations of Tf, and of a large series of simultaneously collected blood and hemodynamic variables, were obtained in 17 patients with post-traumatic sepsis. Tf during sepsis was consistently low (mean +/- SD = 1.46 +/- 0.46) however fluctuated markedly according to changes in metabolic and hemodynamic patterns. Regression analysis showed that decreases in Tf were simultaneously correlated with the plasma lipid pattern (in particular with decreasing cholesterol and increasing triglycerides), with decreases in albumin and peripheral O2 extraction, and with increasing cardiac index (p < 0.001 for all). Decreases in Tf were moderated by increasing the parenteral amino acid dose (p < 0.001). Combinations of these variables in multiple regressions explained nearly 80% of the variability of Tf. There were no similar correlations for other acute phase proteins except ceruloplasmin, which showed opposite changes compared to those of Tf. These results show that within the hypotransferrinemia which characterizes sepsis, Tf may oscillate remaining strongly correlated with changes in metabolic and hemodynamic patterns, which may account for nearly 80% of the variability of Tf. PMID- 18392772 TI - Solid phase synthesis of peptides containing novel amino acids, substituted 3 benzimidazolealanines. AB - A direct solid-phase synthesis of a series of substituted benzimidazole containing peptides is described. The method involves on-resin formation of new amino acids containing benzimidazole derivatives in the side chain. The heterocycle conjugates were obtained by reaction between aldehydes and peptides containing beta-(3,4-diaminophenyl)alanine residue, immobilized on a polymeric solid support. PMID- 18392773 TI - Anti-glomerular basement membrane antibody disease in Japan: part of the nationwide rapidly progressive glomerulonephritis survey in Japan. AB - Anti-glomerular basement membrane (anti-GBM) antibody disease is a rare, but well characterized cause of glomerulonephritis. It is defined by the presence of autoantibodies directed at specific antigenic targets within the glomerular basement membrane. This pattern of rapidly progressive glomerulonephritis and alveolar hemorrhage is often referred to as Goodpasture's syndrome. The prognosis for patients with anti-GBM antibody disease is poor. In Japan, to improve the prognosis of patients with rapidly progressive glomerulonephritis (RPGN), we conducted a nationwide survey of patients with RPGN and investigated the initial symptoms, laboratory findings including renal biopsy findings, treatment methods, and outcomes. Among patients with RPGN, patients with anti-GBM antibody disease were rare: 6.6% (47/715). Alveolar hemorrhage (Goodpasture's syndrome) was observed in 23.4% of patients with anti-GBM antibody disease. Most patients with anti-GBM antibody disease had renal failure at the time of diagnosis. The mean serum creatinine level of patients with renal-limited anti-GBM antibody disease was 7.07 +/- 4.21 mg/dl and that of patients with Goodpasture's syndrome was 7.99 +/- 4.31 mg/dl. The mean level of crescent formation was 78.99 +/- 23.54% in patients with anti-GBM antibody disease, and a cellular crescent form was observed in 63.2% of those patients. The prognosis for patients with anti-GBM antibody disease is poor; the renal survival rate at 6 months after onset was 20.9%, and the mortality at 6 months after onset was 23.3%. To improve the prognosis for anti-GBM antibody disease, it may be necessary to detect this disease in the early stages and to treat it without delay. PMID- 18392774 TI - Endolymphatic sac papillary tumor. PMID- 18392775 TI - Late onset cerebellar metastasis from oesophageal adenocarcinoma in Williams Syndrome. PMID- 18392776 TI - Metastases to the cerebellum. Results and prognostic factors in a consecutive series of 44 operated patients. AB - BACKGROUND: Recent reports on large number of patients with brain metastases report that Whole Brain Radiotherapy (WBRT) and Radiosurgery (RS) should be the treatments of choice, particularly in multiple lesions cases. Among the prognostic factors, the cerebellar location was never considered, although this results in hydrocephalus, brain stem compression, ataxia, intracranial hypertension. MATERIALS AND METHODS: We evaluated 44 patients with cerebellar metastases operated over 6 years. Primary lesions were: Lung (15), Breast (12), Gastrointestinal (9), Gut (3), Ovary (2), Melanoma (1), Salivary gland carcinoma (1), Unknown (1). Lesions were <3 cm in 11 cases, > or =3 cm in 33. Average KPS scoring at admission was 69.9. Twenty nine scored > or =70, 15 < 70. RESULTS: Two patients died for surgical complications, 2 died within 1 months for other causes, 2 were lost to follow up. Eight had postoperative hematoma requiring reoperation, 1 had an occipital infarction. Average KPS scoring at discharge was 76.4, P < 0.002. Those patients that had complications scored less, the difference is significant (P < 0.008). Median survival was 8 months, 1 year survival rate 29.9%. Survival was correlated with either admission or discharge KPS (> or =70 vs. <70): P = 0.05 and P = 0.0001 respectively. None of the other parameters considered reached statistical significance. CONCLUSIONS: Open microneurosurgery is probably still the most effective therapy in improving survival and KPS in patients with large cerebellar metastases, given that the proper surgical technique is used and that complications do not occur. Specific data on cerebellar metastases as an independent subgroup are needed from radiosurgical series. PMID- 18392777 TI - A lectin receptor-like kinase is required for pollen development in Arabidopsis. AB - Lectin receptor-like kinases (Lectin RLKs) are a large family of receptor-like kinases with an extracellular legume lectin-like domain. There are approximately 45 such receptor kinases in Arabidopsis thaliana. Surprisingly, although receptor like kinases in general are well investigated in Arabidopsis, relatively little is known about the functions of members of the Lectin RLK family. A number of studies implicated members of this family in various functions, such as disease resistance, stress responses, hormone signaling, and legume-rhizobium symbiosis. Our current work demonstrated that mutation in one Lectin RLK gene led to male sterility in Arabidopsis. The sterility was due to defects in pollen development. Pollen development proceeded normally in the mutant until anther stage 8. After that, all pollen grains deformed and collapsed. Mature pollen grains were much smaller than wild-type pollen grains, glued together, and totally collapsed. Therefore, the mutant was named sgc, standing for small, glued-together, and collapsed pollen mutant. The mutant phenotype appeared to be caused by an unidentified sporophytic defect due to the mutation. As revealed by analysis of the promoter-GUS transgenic plants and the gene expression analysis using RT-PCR, the gene showed an interesting temporal and spatial expression pattern: it had no or a low expression in young flowers (roughly before anther stage 6), reached a maximum level around stages 6-7, and then declined gradually to a very low level in young siliques. No expression was detected in microspores or pollen. Together, our data demonstrated that SGC Lectin RLK plays a critical role in pollen development. PMID- 18392778 TI - Sucrose induction of Arabidopsis miR398 represses two Cu/Zn superoxide dismutases. AB - MicroRNAs (miRNAs) are approximately 21-nt RNAs that reduce target accumulation through mRNA cleavage or translational repression. Arabidopsis miR398 regulates mRNAs encoding two copper superoxide dismutase (CSD) enzymes and a cytochrome c oxidase subunit. miR398 itself is down-regulated in response to copper and stress. Here we show that miR398 is positively regulated by sucrose, resulting in decreased CSD1 and CSD2 mRNA and protein accumulation. This sucrose regulation is maintained both in the presence and absence of physiologically relevant levels of supplemental copper. Additionally, we show that plants expressing CSD1 and CSD2 mRNAs with altered miR398 complementarity sites display increased mRNA accumulation, whereas CSD1 and CSD2 protein accumulation remain sensitive to miR398 levels, suggesting that miR398 can act as a translational repressor when target site complementarity is reduced. These results reveal a novel miR398 regulatory mechanism and demonstrate that plant miRNA targets can resist miRNA regulation at the mRNA level while maintaining sensitivity at the level of protein accumulation. Our results suggest that even in plants, where miRNAs are thought to act primarily through target mRNA cleavage, monitoring target protein levels along with target mRNA levels is necessary to fully assess the consequences of disrupted miRNA-mRNA pairing. Moreover, the limited complementarity required to maintain robust miR398-directed repression of target protein accumulation suggests that similarly regulated endogenous plant miRNA targets may have eluded detection. PMID- 18392779 TI - A phosphatidylinositol phosphate-specific myo-inositol polyphosphate 5 phosphatase required for seedling growth. AB - The phosphatidylinositol phosphate signaling pathway is involved in many crucial cellular functions. The myo-inositol polyphosphate 5-phosphatases (5PTases) (E.C. 3.1.3.56) comprise a large protein family that hydrolyze 5-phosphates from a variety of phosphatidylinositol phosphate and inositol phosphate substrates. We previously reported that the At5PTase11 enzyme (At1g47510), which is one of the smallest predicted 5PTases found in any organism, encodes an active 5PTase whose activity is restricted to tris- and bis-, but not mono-phosphorylated phosphatidylinositol phosphate substrates containing a 5-phosphate. This is in contrast to other unrestricted Arabidopsis 5PTases, which also hydrolyze tris- and bis inositol phosphate molecules. To further explore the function of At5PTase11, we have characterized two T-DNA mutants in the At5PTase11 gene, and have complemented this mutant. Seed from 5ptase11 mutants germinate slower than wildtype seed and mutant seedlings have decreased hypocotyl growth as compared to wildtype seedlings when grown in the dark. This phenotype is the opposite of the increased hypocotyl growth phenotype previously described for other 5ptase mutants defective in inositol phosphate-specific 5PTase enzymes. By labeling the endogenous myo-inositol pool in 5ptase11 mutants, we correlated these hypocotyl growth changes with a small increase in the 5PTase11 substrate, phosphatidylinositol (4,5) bisphosphate, and decreases in the potential products of 5PTase11, phosphatidylinositol (3) phosphate and phosphatidylinositol (4) phosphate. Surprisingly, we also found that dark-grown 5ptase11 mutants contain increases in inositol (1,4,5) trisphosphate and an inositol bisphosphate that is not a substrate for recombinant 5PTase11. We present a model for regulation of hypocotyl growth by specific molecules found in this pathway. PMID- 18392780 TI - Lack of awareness of oesophageal carcinoma among the public in Ireland. AB - BACKGROUND: Oesophageal cancer is advanced in the majority at presentation and its symptoms are usually present for many months suggesting poor awareness of its symptoms. Few studies have examined awareness of oesophageal cancer amongst the public. AIMS: This study aimed to identify the level of awareness among the general public of oesophageal cancer, of its symptoms, of its awareness campaigns and to compare it with other common cancers. METHODS: Face-to-face interviews were conducted with 279 members of the public. People were asked about their awareness of a range of cancers, and their knowledge of cancer symptoms and cancer awareness campaigns. RESULTS: Awareness of oesophageal cancer was low and knowledge of its symptoms was even lower. Despite the efforts of awareness campaigns, knowledge of these campaigns was poor amongst the public. CONCLUSION: Awareness of oesophageal cancer and its symptoms is low amongst the public. This needs to be addressed if disease is to be detected at an earlier and curable stage. PMID- 18392781 TI - A review of 15-year experience with anomalous origin of the left coronary artery. AB - BACKGROUND: Anomalous left coronary artery from the pulmonary artery (ALCAPA) is a rare condition presenting in infancy with acute myocardial ischaemia. AIMS: A retrospective study was undertaken to assess the presenting features and long term outcome of all cases of ALCAPA presenting to our institution over a 15-year period. METHODS: All the cases were located using the hospital discharge system and charts were reviewed. RESULTS: The mean age at diagnosis was 4.5 months (2 weeks to 16 months). The predominant presenting symptoms were irritability, pallor, and tachypnoea. The predominant electrocardiogram findings were Q waves and ST segment changes in the anterolateral chest leads. One died from the 11 patients in our series. CONCLUSION: The outcome for surgical re-implantation in infants with a diagnosis of ALCAPA is very good however, early diagnosis is crucial to survival. Although once successfully repaired, patients in general were free of symptoms, ventricular dysfunction was usually present, requiring long-term follow-up. PMID- 18392782 TI - Sacral nerve stimulation for faecal incontinence. AB - BACKGROUND: Faecal incontinence resulting from obstetric injury is a socially disabling condition with a significant impact on quality of life. Sacral nerve stimulation (SNS) is a relatively new treatment modality, which offers patients a potential for improved continence. METHODS: We reviewed our initial experience of SNS in 14 patients (mean age 54 years, range 30-72) with faecal incontinence from January 2006 to June 2007. Background demographics, past medical and obstetric history, anal manometry, endoanal ultrasound and pudendal nerve studies were recorded on all patients. All patients who had permanent SNS implants inserted had pre and post operative questionnaires consisting of the Wexner Continence Score and the Rockwood and SF-36 Quality of Life Indices. RESULTS: Out of 14 patients, 13 had incontinence related to obstetric injuries while one was related to a cauda equina syndrome. All patients had a test procedure consisting of placement of temporary electrodes and a 2-week trial of external SNS. Ten patients noted a significant improvement in their continence and these 10 patients subsequently had a permanent SNS device implanted with an overall significant improvement in continence (P < 0.001) and quality of life (P < 0.01). There were no immediate postoperative complications and one late failure consisting of a lead fracture, which was replaced successfully. Four (29%) patients did not have a significant benefit from temporary SNS and two of these patients subsequently had a colostomy. CONCLUSIONS: SNS offers improvement in continence and quality of life in patients with faecal incontinence whose only other option might otherwise be a permanent colostomy. PMID- 18392784 TI - Computer-assisted volumetric analysis compared with ABC/2 method for assessing warfarin-related intracranial hemorrhage volumes. AB - INTRODUCTION: Intraparenchymal hemorrhage (IPH) volume is a powerful predictor of 30-day mortality. Warfarin-related intracranial hemorrhage (ICH) has a higher mortality than ICH without anticoagulation, possibly due to continued growth after 24 h, larger average size, and extension to extraparenchymal compartments. We compared 2 methods of measuring ICH volume in patients with warfarin-related ICH. METHODS: ICH volume was estimated using the ABC/2 method and a computer assisted method (Analyze 6.0 software) applied to the initial head computed tomographic scans in a consecutive series of 8 patients with warfarin-related ICH. The 2 methods were compared for relative and absolute differences in estimated hematoma volumes. RESULTS: The ABC/2 method underestimated hematoma volume in 4 of 5 patients with IPH, inaccurately calculated intraventricular hemorrhage volume in 2 patients (overestimated by 9%, underestimated by 23%), and underestimated a complex subdural hematoma in 1 patient by 24% despite use of the Gebel modification. The mean percentage difference between the Analyze and ABC/2 methods was 24% in ellipsoid hemorrhages and 28% in nonellipsoid hemorrhages (P = 0.77). The mean of the absolute difference between the 2 methods was 6.7 cm3 in ellipsoid hemorrhages and 38.0 cm3 in nonellipsoid hemorrhages (P = 0.18). CONCLUSION: The ABC/2 method accurately and quickly estimates smaller, ellipsoid intraparenchymal hematomas but is inaccurate for larger, complex-shaped warfarin related intraparenchymal, intraventricular, and subdural hematomas. Warfarin related ICH mortality may be underestimated by the ABC/2 method because of larger, complex-shaped, and multicompartmental hematomas. PMID- 18392785 TI - Decompressive Craniectomy. AB - Decompressive Craniectomy (DC) is used to treat elevated intracranial pressure that is unresponsive to conventional treatment modalities. The underlying cause of intracranial hypertension may vary and consequently there is a broad range of literature on the uses of this procedure. Traumatic brain injury (TBI), middle cerebral artery (MCA) infarction, and aneurysmal subarachnoid hemorrhage (SAH) are three conditions for which DC has been predominantly used in the past. Despite an increasing number of reports supportive of DC, the controversy over the suitability of the procedure and criteria for patient selection remains unresolved. Although the majority of published studies is retrospective, the recent publication of several randomized prospective studies prompts a reevaluation of the utility of DC. We review the literature concerning the use of DC in TBI, MCA infarction, and SAH and address the evidence regarding common questions pertaining to the timing of and laterality of the procedure. We conclude that at the time of this review, there still remains insufficient data to support the routine use of DC in TBI, stroke or SAH. There is evidence that early and aggressive use of DC in good-grade patients may improve outcome, but the notion that DC is indicated in these patients is contentious. At this point, the indication for DC should be individualized and its potential implications on long-term outcomes should be comprehensively discussed with the caregivers. PMID- 18392786 TI - Angiotensin II- and glucose-stimulated extracellular matrix production: mediation by the insulin-like growth factor (IGF) axis in a murine mesangial cell line. AB - In diabetic nephropathy, glomerular mesangial cells exhibit aberrant anabolic activity that includes excessive production of extracellular matrix (ECM) proteins, leading to crowding of filtration surface areas and possible renal failure. In the present study, a murine mesangial cell line (MES-13 cells) was studied to determine the roles of the renin-angiotensin system (RAS) and the insulin-like growth factor (IGF) axis in the anabolic response to elevated glucose levels. Culture of MES-13 cells in medium containing supra-physiological glucose concentrations (>5.5 mmol/l) resulted in increased production of ECM proteins including laminin, fibronectin, and heparan sulfate proteoglycan with concurrent increases in IGF-binding protein (IGFBP)-2 production. These responses were blocked by the angiotensin receptor antagonists saralasin and losartan, while exogenous angiotensin II (Ang II) treatment directly stimulated increases in ECM and IGFBP-2. In all experiments, IGFBP-2 levels were correlated with anabolic activity implicating IGFBP-2 as a possible mediator in cellular responses to high glucose and Ang II. Such mediation appears to involve IGFBP-2 modulation of IGF-I signaling, since all responses to high glucose or Ang II were blocked by immuno-neutralization of IGF-I. These data suggest alterations in the IGF axis as key mechanisms underlying nephropathic responses of mesangial cells to Ang II and high glucose. PMID- 18392789 TI - Serial plasma BNP levels in assessing inadequate left ventricular decompression on ECMO. AB - We describe a female infant with complex single ventricle physiology who had undergone median sternotomy for placement of a right-sided systemic-to-pulmonary artery shunt, division of a patent ductus arteriosus, and left pulmonary artery augmentation. Her early postoperative course was complicated by cardiac arrest requiring institution of extracorporeal membrane oxygenation (ECMO) support. The brain natriuretic peptide (BNP) levels acutely improved after left ventricular decompression by insertion of a left atrial cannula. In this setting BNP levels may be an indicator of left ventricular stretch and are potentially a useful index to monitor left ventricular distension. PMID- 18392787 TI - Placental vascularity and growth factor expression in singleton, twin, and triplet pregnancies in the sheep. AB - For singleton, twin, and triplet pregnancies, uteri were collected on day 140 of pregnancy. For each ewe (n = 18), placentomes were fixed by arterial perfusion supplying the fetal (cotyledon) and maternal placenta (caruncle). Tissue sections were stained for determination of vascularity by image analysis. Further, protein expression for factor VIII, vascular endothelial growth factor (VEGF) and its receptor, VEGFR1, as well as basic fibroblast growth factor (FGF2) and its receptor, FGFR, in tissue sections was determined by immunohistochemistry and image analyses. Cotyledonary and caruncular samples were analyzed for expression of mRNA for Vegf and its two receptors, Vegfr1 and Vegfr2, as well as Fgf2 and Fgfr. Fetal number did not affect placental capillary density or factor VIII expression, whereas increased fetal number reduced total cotyledon and caruncle capillary volume. While expression of Vegf, Vegfr1, Vegfr2, and Fgfr mRNA in cotyledonary but not caruncular tissue was greater in twin pregnancies compared to singleton and triplet pregnancies, protein expression of VEGF in the placentome decreased with increasing numbers of fetuses, VEGFR1 did not change, and FGFR was greater in twin versus singleton and triplet pregnancies. Fetal number did not affect the expression of Fgf2 mRNA in placental tissues, whereas FGF2 protein expression was less in triplet compared to singleton and twin pregnancies. Reduced fetal and placental weights in twins and/or triplet pregnancies are associated with an overall decrease in total placental vascularity, VEGF and FGF2 and/or FGFR protein expression, but not in angiogenic factor mRNA expression or VEGFR1 protein expression in sheep. PMID- 18392788 TI - Aminoterminal brain natriuretic peptide levels in children and adolescents after Fontan operation correlate with congestive heart failure. AB - The aim of this study was to generate normal values of amino-terminal pro-brain natriuretic peptide (NT-pro-BNP) in children and adolescents after Fontan operation without congestive heart failure (CHF) and to test the hypothesis that plasma levels of NT-pro-BNP correlate with the clinical severity of CHF. NT-pro BNP plasma levels of 59 consecutive patients, with a median age of 8.4 years, after Fontan operation were measured using an automated enzyme immunoassay. The 97.5th percentile of NT-pro-BNP in patients without CHF was 282.3 pg/ml. The severity of heart failure was quantified by a pediatric cardiologist using the New York University Pediatric Heart Failure Index (NYUPHFI). NT-pro-BNP levels correlated with the NYUPHFI (p = 0.001). In patients with CHF (14/59) the NT-pro BNP levels were significantly higher (median, 399 pg/ml; range, 140-5440 pg/ml) than in patients without CHF (median, 96 pg/ml; range, 11-376 pg/ml). NT-pro-BNP levels of patients with Fontan circulation without CHF are similar to those of healthy children. Plasma NT-pro-BNP concentrations correlate with the severity of CHF in children and adolescents after Fontan operation. Plasma NT-pro-BNP levels can help clinicians in the detection of CHF in pediatric patients with Fontan circulation. PMID- 18392790 TI - Transcatheter management of subtotal cor triatriatum sinister: a rare anomaly. AB - Subtotal cor triatriatum sinister (partially divided left atrium) is characterized by a membrane dividing the left atrium into two portions: a dorsal accessory proximal chamber that receives part of the pulmonary venous return and a small restrictive defect in the membrane that obstructs blood flow from the accessory chamber to the left atrium and results in unilateral pulmonary venous hypertension. This clinical entity is very rarely reported in the literature. This first report of successful transcatheter management of a subdivided left atrium obstructing right pulmonary venous drainage discusses the anatomic, morphologic, and procedural details. PMID- 18392791 TI - Approaches to the treatment of anaemia in patients with chronic heart failure. AB - An association between anaemia, poor functional status and, compared to non anaemic patients, worse clinical status and a higher risk of hospitalisation and death has been consistently reported in chronic heart failure (CHF), although cause an effect has not been proven. While it is attractive to think that correction of a co-morbidity that exacerbates already diminished delivery of oxygen to the tissues in heart failure is likely to beneficial, the possible haemodynamic effects of increasing haemoglobin, for example vasoconstriction, might not be. Consequently, the balance of benefit and risk of anaemia correction in CHF is uncertain, may vary according to the severity of anaemia (and other factors) and needs to be properly evaluated. To date, most studies of anaemia correction in CHF have used erythropoiesis stimulating agents (ESAs). The trials with erythropoietin have been of small size, uncontrolled or unblended/single blind, raising concerns again about interpretation of subjective outcomes. In addition, the analyses of these trials have been suboptimal. Two double-blind, placebo-controlled, darbepoetin studies have been published in full. Neither showed an improvement in functional capacity or consistent effect on patient reported symptoms/quality of life. Darbepoetin is, however, currently being tested in a large-scale, phase III morbidity and mortality trial, the Reduction of Events with Darbepoetin alfa in Heart Failure (RED-HF) which should contribute important information of the safety and efficacy of ESAs in this syndrome. Other approaches, notably parenteral iron supplementation, are also being evaluated and other agents for anaemia correction are under development. PMID- 18392792 TI - [Stage-adjusted treatment for haemorrhoidal disease]. AB - Haemorrhoidal disease is one of the most frequent disorders in western countries. The aim of individual therapy is eradication of symptoms achieved by normalisation of anatomy and physiology. Treatment is orientated to the stage of the disease: First-degree haemorrhoids are treated conservatively. In addition to high fibre diet, sclerotherapy is used. Haemorrhoids of the 2nd degree prolapse during defecation and return spontaneously. First-line treatment is rubber band ligation. Third-degree haemorrhoids that prolapse during defecation have to be digitally reduced. The majority of these patients need surgery. For segmental disorders haemorrhoidectomy according to Milligan-Morgan or Ferguson is recommended. In circular disease Stapler haemorrhoidopexy is now the procedure of choice. Using a classification orientated therapeutical regime orientated to the classification of haemorrhoidal disease offers high healing rates with a low rate of complications and recurrences. PMID- 18392793 TI - [Should surface photodynamic therapy be used for skin neoplasms]. PMID- 18392794 TI - R Regulation of tumor angiogenesis and metastasis by FGF and PDGF signaling pathways. AB - In a fast-growing malignant tissue, tumor blood vessels are exposed to multiple growth factors and cytokines. Although the role of individual factors and their signaling pathways in regulation of tumor neovascularization is relatively well studied, little is known about complex interactions between these factors and their cooperative effects in promoting tumor angiogenesis and metastasis. Our recent studies show that quiescent vascular endothelial cells usually remaining silence to platelet-derived growth factor (PDGF)-BB stimulation acquire their hyperresponsiveness after stimulation with fibroblast growth factor (FGF)-2, which transcriptionally switches on PDGF receptor expression in the activated endothelial cells. Interestingly, PDGF-BB also transduces positive feedback signals to the FGF-2 signaling system by amplifying its receptor expression in vascular mural cells. These uncoordinated reciprocal interactions in the tumor environment lead to the formation of disorganized and primitive vasculatures that facilitate tumor growth and metastasis in mice. These findings provide an example of complex interaction between tumor angiogenic factors. Thus, therapeutic development of antiangiogenic agents for the treatment of cancer should be aimed to block multiple angiogenic signaling pathways and their interactive loops. PMID- 18392795 TI - Stenusine, an antimicrobial agent in the rove beetle genus Stenus (Coleoptera, Staphylinidae). AB - Stenusine is well known as the alkaloid, discharged by the rove beetle, genus Stenus Latreille (Coleoptera, Staphylinidae). The Stenus beetles employ the alkaloid as an escape mechanism when on water surfaces. In the case of danger, they lower their abdomen and emit stenusine from their pygidial glands. Stenusine shows a low surface tension and therefore a high spreading pressure; these properties propel the beetle quickly over the water. Many Steninae do not live in habitats with open waters, but in detritus, leaf litter, mosses, etc. This raises the possibility that stenusine might also have another function, e.g., as antibiotic or fungicide. Stenus beetles show an intense grooming behaviour. With gas chromatography-mass spectrometry analyses we could prove that they cover themselves with their secretion. To tests its antimicrobial properties we conducted agar diffusion tests with stenusine and norstenusine, another substance that is abundant in most Stenus species. Both compounds have an antimicrobial effect on entomopathogenic bacteria and fungi. Stenusine not only allows for an extraordinary method of locomotion on water surfaces, it also protects the Steninae from being infested with microorganisms. PMID- 18392796 TI - Sexual harassment and female gregariousness in the South American sea lion, Otaria flavescens. AB - Most colonial pinnipeds form extreme clusters of breeding females that cannot be entirely explained by the distribution of sites for reproduction. Avoidance of male harassment has been postulated as an important determinant of reproductive aggregation in this group of mammals. Female gregariousness can reduce harassment by resident males by two mechanisms; directly by the 'dilution effect' or indirectly because resident males that defend large female groups are less harassing. In order to investigate the relationship between male harassment and female gregariousness in relation to the size of breeding groups, we analysed the behaviour of dominant males and their females in a breeding colony of Otaria flavescens. Females in large breeding groups received less harassment by resident males due to dilution effects and because males that defended a large group interacted less frequently with females than males with small groups. PMID- 18392797 TI - [Increased sensitivity to pain in long-term opioid treatment]. AB - Recent years have seen a rise in the prescription of strong opioids for chronic and even subthreshold forms of pain. Animal and human experiments and clinical observations have shown that, compared to placebos, chronic opioid administration results in not only tolerance to its analgesic effects but also in heightened pain sensitivity. Therefore chronic, especially high-dose, opioid treatment can not be recommended for chronic pain but is instead contraindicated. Patients on long-term opioids will often reject proposals to withdraw the drug. Important elements of treatment are patient education, empathy and unconditional acceptance by the patient, motivation enhancement and psychological support, collaboration with other prescribers and therapists, detoxification, measures to increase pain tolerance, encouragement of general health behavior and salutotherapy, treatment of the primary illness, and prescription of participation oriented alternative analgesics. PMID- 18392798 TI - [Detection of papillomavirus DNA in the prostate: a virus with underestimated clinical relevance?]. AB - BACKGROUND: Human papillomaviruses (HPV) are the most frequent pathogens of sexually transmitted diseases. They have been associated with an increased incidence of several anogenital tumors. Whether oncogenic HPV are involved in the pathogenesis of prostate cancer has been a subject of great controversy. This study's purpose was to investigate the association between HPV infection and prostate cancer (PCA). MATERIAL AND METHODS: The study included 213 consecutive patients with an average age of 65.7 (+/-8.4) years. Within the framework of transrectal, ultrasonic-guided multibiopsy of the prostate, one additional core was examined by means of polymerase chain reaction (PCR) in relation to bacterial, fungal, and viral (including HPV) DNA, with subsequent DNA sequencing. The collected data were correlated with the histological results and with diverse clinical variables. The influence of several predictors for the existence of PCA was verified with a logistic regression model. RESULTS: No general bacterial DNA (16S rDNA) was detected. Of the 213 patients, 145 (68.1%) showed HPV DNA. In 64% (n=137), high-risk HPV DNA were depicted; these were 18% of the total in each case of HPV genotypes 16 and 18. From our examinations, no significant positive correlation existed between the HPV evidence and the histologically verified PCA that was found in 23.5% of the patients (n=50; odds ratio 1.45; 95% confidence interval 0.71-2.91). The BK virus was not found in any of the cores confirmed through PCR. CONCLUSION: Although no positive correlation between HPV infection and PCA existed in our study, data from the literature suggest an influence of the papillomavirus on PCA oncogenesis. Future studies should highlight to what extent HPV DNA is inserted in the genome of prostate cells and is able to cause subsequent malignant transformation of particular genes. PMID- 18392799 TI - Transfer of anthracnose resistance and pod coiling traits from Medicago arborea to M. sativa by sexual reproduction. AB - Five asymmetric hybrid plants were obtained between Medicago sativa (2n = 4x = 32) and Medicago arborea (2n = 4x = 32) through sexual reproduction and the use of a cytoplasmically male sterile M. sativa genotype. Over 2,000 pollinations were made to obtain these hybrids. Amplified fragment length polymorphism (AFLP) analysis showed that in the most studied hybrid (WA2273), 4% of the bands unique to the M. arborea parent were present, versus 72% for the unique M. sativa bands. This suggests that only a single M. arborea chromosome or chromosome parts has been transferred. WA2273 had 7% of AFLP bands which were not present in either parent, which is suggestive of chromosome rearrangements as would be expected if only chromosome parts or a single part had been transferred from M. arborea. Phenotypic evidence for hybridity was obtained for pod coiling (1.4 coils in WA2273 versus three coils in the M. sativa parent and its self and testcross populations, and one coil in M. arborea), and Colletotrichum trifolii race 2 resistance (transferred from the resistant M. arborea parent, as the M. sativa parent and the self populations were highly susceptible). The hybrids were self sterile, but were female fertile to a high level when crossed with 4x, but not 2x, M. sativa, indicating they were at or near 4x. Both the pod coiling trait and anthracnose resistance segregated in the progeny of testcrosses between WA2273 and M. sativa. The work demonstrates that agronomically useful traits can be introgressed into M. sativa from M. arborea by use of male sterile M. sativa and sexual reproduction. PMID- 18392800 TI - Molecular mapping of the novel powdery mildew resistance gene Pm36 introgressed from Triticum turgidum var. dicoccoides in durum wheat. AB - Powdery mildew, caused by Blumeria graminis f.sp. tritici, is one of the most important wheat diseases in many regions of the world. Triticum turgidum var. dicoccoides (2n=4x=AABB), the progenitor of cultivated wheats, shows particular promises as a donor of useful genetic variation for several traits, including disease resistances. The wild emmer accession MG29896, resistant to powdery mildew, was backcrossed to the susceptible durum wheat cultivar Latino, and a set of backcross inbred lines (BC(5)F(5)) was produced. Genetic analysis of F(3) populations from two resistant introgression lines (5BIL-29 x Latino and 5BIL-42 x Latino) indicated that the powdery mildew resistance is controlled by a single dominant gene. Molecular markers and the bulked segregant analysis were used to characterize and map the powdery mildew resistance. Five AFLP markers (XP43M32((250)), XP46M31((410)), XP41M37((100)), XP41M39((250)), XP39M32((120))), three genomic SSR markers (Xcfd07, Xwmc75, Xgwm408) and one EST-derived SSR marker (BJ261635) were found to be linked to the resistance gene in 5BIL-29 and only the BJ261635 marker in 5BIL-42. By means of Chinese Spring nullisomic tetrasomic, ditelosomic and deletion lines, the polymorphic markers and the resistance gene were assigned to chromosome bin 5BL6-0.29-0.76. These results indicated that the two lines had the same resistance gene and that the introgressed dicoccoides chromosome segment was longer (35.5 cM) in 5BIL-29 than that introgressed in 5BIL-42 (less than 1.5 cM). As no powdery mildew resistance gene has been reported on chromosome arm 5BL, the novel resistance gene derived from var. dicoccoides was designated Pm36. The 244 bp allele of BJ261635 in 5BIL 42 can be used for marker-assisted selection during the wheat resistance breeding process for facilitating gene pyramiding. PMID- 18392801 TI - Soybean molecular linkage group B1 corresponds to classical linkage group 16 based on map location of the lf (2) gene. AB - The seven-leaflet character of soybean [Glycine max L. (Merr.)] is a single recessive trait conditioned by the lf ( 2 ) gene. The lf ( 2 ) gene is located on linkage group (LG) 16 of the classical soybean genetic map, but it has not been placed on the molecular map. The objective of this research was to identify the location of the lf ( 2 ) gene on the soybean molecular map using simple sequence repeat (SSR) markers. A backcross breeding method was used to create three- and seven-leaflet near-isogenic lines in genetic backgrounds of 'Traill', 'MN1401', and 'MN1801'. Eight mapping populations were derived from eight single heterozygous Lf ( 2 ) lf ( 2 ) plants. A total of 482 SSR markers that covered approximately every 10-20 cM of all soybean molecular LG were used to screen the mapping populations for polymorphisms. For the 115 SSRs that were identified as polymorphic, possible linkage between the lf ( 2 ) gene and the polymorphic SSR markers was determined. One SSR marker from the LG B1, Sat_272, was linked (LOD > 4.0) to the lf ( 2 ) gene in the Traill and MN1401 derived populations, with map distances ranging from 2.8 to 11.2 cM. Two additional markers (a SSR, Sat_270 and a SNP, A588c) located on LG B1 were also polymorphic and were identified as linked to the lf ( 2 ) gene in one of the populations. This research was successful in mapping the lf ( 2 ) gene to LG B1 of the soybean molecular map and therefore, provides evidence that molecular LG B1 corresponds to classical LG 16. PMID- 18392802 TI - Molecular mapping of two loci that confer resistance to Asian rust in soybean. AB - Asian soybean rust (ASR) is caused by the fungal pathogen Phakopsora pachyrhizi Sydow & Sydow. It was first identified in Brazil in 2001 and quickly infected soybean areas in several countries in South America. Primary efforts to combat this disease must involve the development of resistant cultivars. Four distinct genes that confer resistance against ASR have been reported: Rpp1, Rpp2, Rpp3, and Rpp4. However, no cultivar carrying any of those resistance loci has been released. The main objective of this study was to genetically map Rpp2 and Rpp4 resistance genes. Two F(2:3) populations, derived from the crosses between the resistant lines PI 230970 (Rpp2), PI 459025 (Rpp4) and the susceptible cultivar BRS 184, were used in this study. The mapping populations and parental lines were inoculated with a field isolate of P. pachyrhizi and evaluated for lesion type as resistant (RB lesions) or susceptible (TAN lesions). The mapping populations were screened with SSR markers, using the bulk segregant analysis (BSA) to expedite the identification of linked markers. Both resistance genes showed an expected segregation ratio for a dominant trait. This study allowed mapping Rpp2 and Rpp4 loci on the linkage groups J and G, respectively. The associated markers will be of great value on marker assisted selection for this trait. PMID- 18392803 TI - Influence of pregnancy on long-term progression of retinopathy in patients with type 1 diabetes. AB - AIMS/HYPOTHESIS: Pregnancy in type 1 diabetic women is associated with risk of worsening of retinopathy. It has been reported that deterioration continues in the months after delivery, but direct data are lacking. It is also unclear what impact pregnancy has on the long-term progression of retinopathy. METHODS: We studied 59 women with type 1 diabetes who had retinal photographs before pregnancy and yearly for 5 years post pregnancy. These photographs were graded using the EURODIAB retinopathy grading system. RESULTS: The mean duration of diabetes was 14.4+/-8.2 years and mean age at pregnancy was 29.8+/-5.5 years. Mean HbA(1c) was 8.2+/-2.0% before pregnancy with tighter control during pregnancy itself. This value was high despite efforts to improve take-up of pre conception care. Mean HbA(1c) was 8.6+/-1.5 during the follow-up period. At baseline, 43 (72.9%) women were free of retinopathy, 15 had non-proliferative retinopathy and one woman had previously had laser therapy. During pregnancy four women required laser therapy. Over the next 5 years none required laser therapy, although retinopathy worsened in 14 women. Ten-year follow-up data were available on 22 women, one of whom required laser therapy 8 years after pregnancy. Baseline retinopathy status was the only independent risk factor which predicted progression of retinopathy. CONCLUSIONS/INTERPRETATION: Pregnancy is not associated with post-partum worsening of retinopathy. PMID- 18392805 TI - A dead heat: target normal glucose levels in the intensive care unit--but with caution. PMID- 18392804 TI - Novel biochemical risk factors for type 2 diabetes: pathogenic insights or prediction possibilities? AB - This review critically appraises studies examining the association of novel factors with diabetes. We show that many of the most studied novel and apparently 'independent' risk factors are correlated with each other by virtue of their common origins or pathways, and that residual confounding is likely. Available studies also have other limitations, including differences in methodology or inadequate statistical analyses. Furthermore, although most relevant work in this area has focused on improving our understanding of the pathogenesis of diabetes, association studies in isolation cannot prove causality; intervention studies with specific agents (if available) are required, and genetic studies may help. With respect to the potential value of novel risk factors for diabetes risk prediction, we illustrate why this work is very much in its infancy and currently not guaranteed to reach clinical utility. Indeed, the existence of several more easily measured powerful predictors of diabetes, suggests that the additional value of novel markers may be limited. Nevertheless, several suggestions to improve relevant research are given. Finally, we show that several risk factors for diabetes are only weakly associated with the risk of incident vascular events, an observation that highlights the limitations of attempting to devise unified criteria (e.g. metabolic syndrome) to identify individuals at risk of both CHD and diabetes. PMID- 18392806 TI - Biomarkers in the diagnosis of pneumonia in the critically ill: don't shoot the piano player. PMID- 18392807 TI - Triggering receptors expressed on myeloid cells in pulmonary aspiration syndromes. AB - OBJECTIVE: To investigate the potential role of serum and alveolar soluble triggering receptor expressed on myeloid cells (sTREM-1) as a biological marker of pulmonary aspiration syndromes. DESIGN: Prospective cohort study. SETTING: University-affiliated intensive care unit. PATIENTS: Seventy-five patients with pulmonary aspiration and 13 controls receiving mechanical ventilation. INTERVENTIONS: Blood and bronchoalveolar lavage (BAL) fluid samples were collected on enrollment. Soluble TREM-1 levels were measured by an enzyme-linked immunosorbent assay. MEASUREMENTS AND RESULTS: Thirty-eight of 75 participants had documented BAL culture-positive pulmonary aspiration. While circulating levels of sTREM-1 were comparable between those with aspiration syndromes (19.81 +/- 12.09 pg/ml) and controls (15.96 +/- 11.16 pg/ml) (p=0.27), the alveolar levels of sTREM-1 were higher in patients with culture-positive pulmonary aspiration (344.41 +/- 152.82 pg/ml) compared with those culture-negative pulmonary aspiration (142.76 +/- 89.88 pg/ml; p < 0.001). A cut-off value of 250 pg/ml for alveolar sTREM-1 achieved a sensitivity of 65.8% (95% CI 48.6-80.4) and a specificity of 91.9% (95% CI 78.1-98.2) with an area under the curve of 0.87 (95% CI 0.78-0.94). CONCLUSIONS: Alveolar sTREM-1 levels can be a potential biomarker for distinguishing BAL culture-positive from BAL culture-negative pulmonary aspiration. PMID- 18392808 TI - ATP-binding cassette transporters in human heart failure. AB - Adenosine triphosphate-binding cassette (ABC) transporters are involved in energy dependent transport of substrates across biological membranes. We hypothesized that their expression is altered during human heart failure, suggesting a pathophysiologic basis. Messenger ribonucleic acid quantification of all known ABC transporters revealed multiple alterations in ABC transporter expression in failing human hearts (New York Heart Association classification III-IV) compared to nonfailing controls. These include a loss of ABCC7 chloride channels and an increased expression of the K(ATP) channel regulatory subunits ABCC8. Moreover, ABCG2, an efflux pump for xenobiotics/drugs, was expressed at much higher levels in failing hearts compared to nonfailing control hearts. ABCG2 was found in cardiac capillary endothelial cells and cardiomyocytes. Experiments in cells stably transfected with human ABCG2 revealed that the peroxisome proliferator activated receptor-gamma agonist rosiglitazone was transported by ABCG2 but also inhibited the export of the prototypical ABCG2 substrate pheophorbide A (IC(50) 25 microM). These results suggest that altered ABC transporter expression in failing hearts might contribute to impaired channel conductance or might affect the cardiac disposition of drugs. PMID- 18392809 TI - Acute inhibitory effect of dronedarone, a noniodinated benzofuran analogue of amiodarone, on Na+/Ca2+ exchange current in guinea pig cardiac ventricular myocytes. AB - Using the whole-cell voltage-clamp method, we examined an acute effect of dronedarone, a noniodinated benzofuran analogue of amiodarone, on Na+/Ca2+ exchange current (INCX) in guinea pig cardiac ventricular cells. The INCX was recorded by ramp pulses with a holding potential of -60 mV using a pipette solution containing 226 nM free Ca2+ (20 mM 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid and 10 mM Ca2+) and 20 mM Na+. The external solution contained 140 mM Na+, 1 mM Ca2+, and blockers of other currents and pumps such as Cs+, nifedipine, ryanodine, and ouabain. A selective potent NCX inhibitor, KB R7943 (100 microM), was used to completely inhibit INCX. Dronedarone inhibited INCX in a concentration-dependent manner. The IC50 values for the outward and inward INCX inhibition were about 33 and 28 microM, respectively, with the Hill coefficient of 1 for both. The inhibitory effect of dronedarone at 50 microM on INCX did not change in the presence of trypsin in the pipette solution. Therefore, dronedarone is classified as a trypsin-insensitive NCX inhibitor and distinct from amiodarone which is a trypsin sensitive. We conclude that dronedarone inhibits INCX but the potency is tenfold less than that of amiodarone. Dronedarone may modestly inhibit INCX in a therapeutic concentration range. PMID- 18392811 TI - Identification of proteins in painting cross-sections by immunofluorescence microscopy. AB - Immunofluorescence microscopy offers a highly specific analytical tool for unambiguous recognition and mapping of proteins in complex matrices. In the present work, the analytical potentials of immunofluorescence microscopy have been exploited to provide recognition of proteinaceous binders in painting cross sections. An optimised analytical protocol is proposed for the identification of ovalbumin and of bovine serum albumin as markers of egg white and casein, respectively. The study has been carried out on laboratory model samples simulating both easel and mural paintings. The obtained results demonstrated the effectiveness of the method, suggesting the potential future use of immunofluorescence microscopy as a routine diagnostic tool in conservation science. Possible developments of the proposed methodology in order to improve the specificity of the method and its detection sensitivity are presented and discussed. PMID- 18392813 TI - Flow-injection solid surface lanthanide-sensitized luminescence sensor for determination of p-aminobenzoic acid. AB - A single optosensing device based on lanthanide-sensitized luminescence was developed for determination of p-aminobenzoic acid (PABA). The method is based on the formation of a complex between PABA and Tb(III) immobilized on the solid phase (QAE A-25 resin) placed inside the flow cell. NaCl (1 M) was used as carrier solution and HCl (0.05 M) as eluent. The sample solutions of PABA (100 MUL) containing Tb(III) and buffered at pH = 6.0 were injected into the carrier stream and the luminescence was measured at lambda (ex) = 290 nm and lambda (em) = 546 nm. The method shows a linear range from 0.2 to 6.0 MUg mL(-1) with an RSD of 1.2% (n = 10) and a sampling frequency of 22 h(-1). A remarkable characteristic of the method is its high selectivity which allows it to be satisfactorily applied to the analysis of PABA in pharmaceutical samples without prior treatment. PMID- 18392814 TI - Quantification with HPLC-MS/MS for environmental issues: quality assurance and quality assessment. PMID- 18392815 TI - Application of at-line two-dimensional liquid chromatography-mass spectrometry for identification of small hydrophilic angiotensin I-inhibiting peptides in milk hydrolysates. AB - A two-dimensional chromatographic method with mass spectrometric detection has been developed for identification of small, hydrophilic angiotensin I-inhibiting peptides in enzymatically hydrolysed milk proteins. The method involves the further separation of the poorly retained hydrophilic fraction from a standard C(18) reversed-phase column on a hydrophilic interaction liquid chromatography (HILIC) column. The latter column is specifically designed for the separation of hydrophilic compounds. Narrow fractions collected from the HILIC column were analysed for their angiotensin I-converting enzyme (ACE) inhibiting potential in an at-line assay. Fractions showing significant inhibition of ACE were analysed by LC-MS for structure elucidation. With this method the main peptides responsible for ACE-inhibition in the hydrophilic part of a milk hydrolysate could be determined. The ACE-inhibiting peptides RP, AP, VK, EK, and EW explained more than 85% of ACE-inhibition by the hydrophilic fraction. PMID- 18392816 TI - Sonographic diagnosis of intestinal polyps in children. AB - Although colonoscopy is the modality of choice for the diagnosis of intestinal polyps, sonography may be the first diagnostic modality in the evaluation of children with this pathology. Graded compression sonography of the small and large bowel may be useful in diagnosing intestinal polyps. A specific diagnosis is possible in many cases as most polyps have a characteristic appearance. Although sonography is not intended to replace endoscopy in the diagnostic work up of intestinal polyps, it is important for radiologists to be aware of this appearance as they can be the first in making the diagnosis of intestinal polyps using sonography. PMID- 18392817 TI - Age-specific effective doses for pediatric MSCT examinations at a large children's hospital using DLP conversion coefficients: a simple estimation method. AB - BACKGROUND: There is a need for an easily accessible method for effective dose estimation in pediatric CT. OBJECTIVE: To estimate effective doses for a variety of pediatric neurological and body CT examinations in five age groups using recently published age- and region-specific dose length product (DLP) to effective dose conversion coefficients. MATERIALS AND METHODS: A retrospective review was performed of 1,431 consecutive CT scans over a 12-week period using age- and weight-adjusted CT protocols. Age- and region-specific DLP to effective dose conversion coefficients were applied to console-displayed DLP data. RESULTS: Effective dose estimates for single-phase head CT scans in neonatal, and 1-, 5-, 10- and 15-year-old age groups were 4.2, 3.6, 2.4, 2.0 and 1.4 mSv, respectively. For abdomen/pelvis CT scans the corresponding effective doses were 13.1, 11.1, 8.4, 8.9 and 5.9 mSv. The range of pediatric CT effective doses is wide, from ultralow dose protocols (<1 mSv) to extended-coverage body examinations (10-15 mSv). CONCLUSION: Age- and region-specific pediatric DLP to effective dose conversion coefficients provide an accessible and user-friendly method for estimating pediatric CT effective doses that is available to radiologists working without medical physics support. PMID- 18392818 TI - A case of severe congenital kyphoscoliosis secondary to multiple bilateral thoracic pedicle aplasia. AB - A 9-month-old boy with congenital kyphoscoliosis secondary to multiple bilateral thoracic pedicle aplasias is presented. This anomaly has rarely been described in the literature. Plain films showed absence of thoracic pedicles bilaterally from T2 to T9, which was better demonstrated on CT multiplanar and 3-D reformatted images. There were no neurological symptoms even though the deformity progressed rapidly. Congenital kyphosis or kyphoscoliosis is often related to spinal anomalies that are located on the anterior aspect of the vertebrae. However, posterior anomalies may also be responsible and should be recognized before neurological complications occur. PMID- 18392819 TI - Spinal osteomyelitis due to Aspergillus flavus in a child: a rare complication after haematopoietic stem cell transplantation. AB - We report the case of a child affected by acute myeloid leukaemia who was treated with allogeneic haematopoietic stem cell transplantation and developed cervicothoracic spinal osteomyelitis due to Aspergillus flavus. The diagnosis was difficult on a clinical basis, but made possible by conventional radiography and MRI. PMID- 18392820 TI - MRI in true hermaphroditism. PMID- 18392821 TI - Secondary hypertrophic osteoarthropathy on bone scintigraphy as a diagnosis of vascular prosthesis infection. PMID- 18392810 TI - Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease: participation of the phospholipase A2 enzyme. AB - RATIONALE: Alzheimer disease (AD), a progressive neurodegenerative disorder, is the leading cause of dementia in the elderly. A combination of cholinergic and glutamatergic dysfunction appears to underlie the symptomatology of AD, and thus, treatment strategies should address impairments in both systems. Evidence suggests the involvement of phospholipase A(2) (PLA(2)) enzyme in memory impairment and neurodegeneration in AD via actions on both cholinergic and glutamatergic systems. OBJECTIVES: To review cholinergic and glutamatergic alterations underlying cognitive impairment and neuropathology in AD and attempt to link PLA(2) with such alterations. METHODS: Medline databases were searched (no date restrictions) for published articles with links among the terms Alzheimer disease (mild, moderate, severe), mild cognitive impairment, choline acetyltransferase, acetylcholinesterase, NGF, NGF receptor, muscarinic receptor, nicotinic receptor, NMDA, AMPA, metabotropic glutamate receptor, atrophy, glucose metabolism, phospholipid metabolism, sphingolipid, membrane fluidity, phospholipase A(2), arachidonic acid, attention, memory, long-term potentiation, beta-amyloid, tau, inflammation, and reactive species. Reference lists of the identified articles were checked to identify additional studies of interest. RESULTS: Overall, results suggest the hypothesis that persistent inhibition of cPLA(2) and iPLA(2) isoforms at early stages of AD may play a central role in memory deficits and beta-amyloid production through down-regulation of cholinergic and glutamate receptors. As the disease progresses, beta-amyloid induced up-regulation of cPLA(2) and sPLA(2) isoforms may play critical roles in inflammation and oxidative stress, thus participating in the neurodegenerative process. CONCLUSION: Activation and inhibition of specific PLA(2) isoforms at different stages of AD could be of therapeutic importance and delay cognitive dysfunction and neurodegeneration. PMID- 18392822 TI - Evaluation of a decision support system for interpretation of myocardial perfusion gated SPECT. AB - PURPOSE: We have recently presented a decision support system for interpreting myocardial perfusion scintigraphy (MPS). In this study, we wanted to evaluate the system in a separate hospital from where it was trained and to compare it with a quantification software package. METHODS: A completely automated method based on neural networks was trained for the interpretation of MPS regarding myocardial ischaemia and infarction using 418 MPS from one hospital. Features from each examination describing rest and stress perfusion, regional and global function were used as inputs to different neural networks. After the training session, the system was evaluated using 532 MPS from another hospital. The test images were also processed with the quantification software package Emory Cardiac Toolbox (ECTb). The images were interpreted by experienced clinicians at both the training and the test hospital, regarding the presence or absence of myocardial ischaemia and/or infarction and these interpretations were used as gold standard. RESULTS: The neural network showed a sensitivity of 90% and a specificity of 85% for myocardial ischaemia. The specificity for the ECTb was 46% (p < 0.001), measured at the same sensitivity. The neural network sensitivity for myocardial infarction was 89% and the specificity 96%. The corresponding specificity for the ECTb was 54% (p < 0.001). CONCLUSION: A decision support system based on neural networks presents interpretations more similar to experienced clinicians compared to a conventional automated quantification software package. This study shows the feasibility of disseminating the expertise of experienced clinicians to less experienced physicians by the use of neural networks. PMID- 18392823 TI - Immunomodulatory drugs Revlimid (lenalidomide) and CC-4047 induce apoptosis of both hematological and solid tumor cells through NK cell activation. AB - Revlimid (Lenalidomide, CC-5013) and CC-4047 are IMiDs immunomodulatory drugs that have been described as having immunomodulatory properties and anti-tumor activity. Here we report proapoptotic effects of CC-5013 and CC-4047 on tumor cells in a co-culture model of PBMC and tumor cells. CC-5013 and CC-4047 enhanced PBMC activity leading to tumor cell apoptosis in K562/PBMC co-culture model. We also demonstrate that the natural killer (NK) cell population of PBMC was essential in inducing K562 apoptosis. Increases of NK and natural killer T (NKT) cell populations by CC-5013 and CC-4047 was observed along with modulation of NK cell CD56 adhesion marker. In addition, our data indicate that NK activation by CC-4047 was dependent on other cell types of PBMC. We expanded the application of K562/PBMC co-culture model to other hematological and solid tumors. In Raji/PBMC co-culture model, CC-5013 and CC-4047 dose-dependently augmented tumor cell apoptosis. Pre-treatment of Raji cells with Rituximab further enhanced apoptosis induced by CC-5013 or CC-4047-treated PBMC. Moreover, CC-5013 and CC-4047 significantly increased PC-3 prostate cancer cell apoptosis in PC-3/PBMC co culture, either as single agent or in combination with Docetaxel. Together, the results reveal that co-culture models are suitable cellular systems to assess anti-tumor activities of these compounds. Our findings support clinical evaluation of CC-5013 and CC-4047 in relapsed NHL with Rituximab and in prostate cancer with Docetaxel. PMID- 18392824 TI - Assessment of immunologic response and recurrence patterns among patients with clinical recurrence after vaccination with a preventive HER2/neu peptide vaccine: from US Military Cancer Institute Clinical Trials Group Study I-01 and I-02. AB - BACKGROUND: E75, a HER2/neu immunogenic peptide, is expressed in breast cancer (BCa). We have performed clinical trials of E75 + GM-CSF vaccine in disease-free, node-positive and node-negative BCa patients at high recurrence risk and recurrences were noted in both control and vaccine groups. METHODS: Among the 186 BCa patients enrolled, 177 completed the study. Patients were HLA typed; the HLA A2(+)/A3(+) patients were vaccinated; HLA-A2(-)/A3(-) patients were followed as controls. Standard clinicopathological factors, immunologic response to the vaccine, and recurrences were collected and assessed. RESULTS: The control group recurrence rate was 14.8 and 8.3% in the vaccinated group (P = 0.17). Comparing the 8 vaccinated recurrences (V-R) to the 88 vaccinated nonrecurrent patients (V NR), the V-R group had higher nodal stage (> or = N2: 75 vs. 5%, P = 0.0001) and higher grade tumors (%grade 3: 88 vs. 31%, P = 0.003). The V-R group did not fail to respond immunologically as noted by equivalent dimer responses and post-DTH responses. Compared to control recurrent patients (C-R), V-R patients trended toward higher-grade tumors and hormone-receptor negativity. C-R patients had 50% bone-only recurrences, compared to V-R patients with no bone-only recurrences (P = 0.05). Lastly, V-R mortality rate was 12.5% compared with 41.7% for the C-R group (P = 0.3). CONCLUSIONS: The vaccinated patients who recurred had more aggressive disease compared to V-NR patients. V-R patients had no difference in immune response to the vaccine either in vitro or in vivo. V-R patients, when compared to C-R patients, trended towards more aggressive disease, decreased recurrence rates, decreased mortality, and no bone-only recurrences. PMID- 18392825 TI - Models of Cheyne-Stokes respiration with cardiovascular pathologies. AB - Cheyne-Stokes respiration (CSR) is a periodic breathing pattern, characterized by short intervals of very little or no breathing (apnea), each followed by an interval of very heavy breathing (hyperpnea). This work presents a new compartmental model of the human cardio-respiratory system, simulating the factors that determine the concentrations of carbon dioxide in the compartments of the cardiovascular system and the lungs. The parameter set on which a Hopf bifurcation gives birth to stable CSR oscillations has been determined. The model predicts that the onset of CSR oscillations may result from an increase in any of: ventilation-perfusion ratio, feedback control gain, transport delay, left heart volume, lung congestion, or cardiovascular efficiency. The model is employed to investigate the relationship between CSR and serious cardiovascular pathologies, such as congestive heart failure and encephalitis, as well as the effects of acclimatization to higher altitudes. In all cases, the model is consistent with medical observations. PMID- 18392826 TI - Traction patterns of tumor cells. AB - The traction exerted by a cell on a planar deformable substrate can be indirectly obtained on the basis of the displacement field of the underlying layer. The usual methodology used to address this inverse problem is based on the exploitation of the Green tensor of the linear elasticity problem in a half space (Boussinesq problem), coupled with a minimization algorithm under force penalization. A possible alternative strategy is to exploit an adjoint equation, obtained on the basis of a suitable minimization requirement. The resulting system of coupled elliptic partial differential equations is applied here to determine the force field per unit surface generated by T24 tumor cells on a polyacrylamide substrate. The shear stress obtained by numerical integration provides quantitative insight of the traction field and is a promising tool to investigate the spatial pattern of force per unit surface generated in cell motion, particularly in the case of such cancer cells. PMID- 18392827 TI - [Malignant odontogenic tumors]. AB - Malignant odontogenic tumors are extremely rare. As with benign odontogenic tumors, malignant epithelial odontogenic tumors or odontogenic carcinomas are distinguished from the even rarer mesenchymal ones, the odontogenic sarcomas. The existence of odontogenic carcinosarcomas is not yet acknowledged by the World Health Organization. Odontogenic carcinomas comprise ameloblastic carcinoma (AmCa), primary intraosseous carcinoma (PIOC), clear cell odontogenic carcinoma, odontogenic ghost cell carcinoma (OGCC), and the special case of metastasizing ameloblastoma. Odontogenic sarcomas consist of ameloblastic fibrosarcoma and ameloblastic fibrodentinosarcoma and fibroodontosarcoma. Whereas metastasizing ameloblastoma can be diagnosed only after having metastasized, all other malignant odontogenic tumors present with atypia, increased cellularity and mitoses, and invasion. Odontogenic sarcomas are regarded as low-grade tumors that rarely metastasize. Odontogenic carcinomas, however, especially AmCa, OGCC, and PIOC, are more aggressive, with a 5-year survival rate of about 70% for AmCa and OGCC and a 3-year survival rate of about 37% for PIOC. Radical surgery, eventually in combination with radiotherapy, is the treatment of choice. PMID- 18392828 TI - [Benign odontogenic ectomesenchymal tumors]. AB - The group of odontogenic ectomesenchymal tumors consists of odontogenic fibroma (epithelium-rich and epithelium-poor types), odontogenic myxoma, and cementoblastoma. Whereas odontogenic fibromas and cementoblastomas are very rare lesions, odontogenic myxoma is the fourth common odontogenic tumor, preceded only by keratocystic odontogenic tumor, the odontomas, and ameloblastoma. The diagnosis of cementoblastoma rests on its connection to the root of a tooth. The differentiation of odontogenic fibroma and myxoma from other lesions, especially from normal structures such as dental follicles and papillae, may be challenging if the X-ray appearance (localized osteolysis containing a tooth) is not appreciated and subtle histological clues (remainders of inner enamel epithelium at the surface of the lesion, dentin fragments) are not properly recognized. While odontogenic fibromas have almost no tendency for recurrence and are treated by enucleation or local excision, cementoblastomas and especially odontogenic myxomas have a high percentage of recurrence if intralesional procedures are applied. Hence, complete resection with free margins is recommended--at least for larger odontogenic myxomas and, especially, lesions in the maxilla--to prevent further extension to the orbita or base of the skull. PMID- 18392829 TI - Low kilovoltage cardiac dual-source CT: attenuation, noise, and radiation dose. AB - The purpose of this study was to investigate the effect of low kilovoltage dual source computed tomography coronary angiography (CTCA) on qualitative and quantitative image quality parameters and radiation dose. Dual-source CTCA with retrospective ECG gating was performed in 80 consecutive patients of normal weight. Forty were examined with a standard protocol (120 kV/330mAs), 20 were examined at 100 kV/330mAs, and 20 at 100 kV/220mAs. Two blinded observers independently assessed image quality of each coronary segment and measured the image parameters noise, attenuation, and contrast-to-noise ratio (CNR). The effective radiation dose was calculated using CT dose volume index and the dose length product. Diagnostic image quality was obtained in 99% of all coronary segments (1,127/1,140) without significant differences among the protocols. Image noise, attenuation, and CNR were significantly higher for 100 kV/330mAs (26 +/- 3 HU, 549 +/- 62 HU, 25.5 +/- 3.2; each P < 0.01) and 100 kV/220mAs (27 +/- 2 HU, 560 +/- 43 HU, 25.0 +/- 2.2; each P < 0.01) when compared to the 120-kV protocol (21 +/- 2 HU, 317 +/- 28 HU, 20.6 +/- 1.7). There was no significant difference between the two 100-kV protocols. Estimated effective radiation dose of the 120 kV protocol (8.9 +/- 1.2 mSv) was significantly higher than the 100 kV/330mAs (6.7 +/- 0.8 mSv, P < 0.01) or 100 kV/220mAs (4.4 +/- 0.6 mSv, P < 0.001) protocols. Dual-source CTCA with 100 kV is feasible in patients of normal weight, results in a diagnostic image quality with a higher CNR, and at the same time significantly reduces the radiation dose. PMID- 18392830 TI - Differences in perfusion parameters between upper and lower lumbar vertebral segments with dynamic contrast-enhanced MRI (DCE MRI). AB - To investigate the influence of age, sex and spinal level on perfusion parameters of normal lumbar bone marrow with dynamic contrast-enhanced MRI (DCE MRI). Sixty seven subjects referred for evaluation of low back pain or sciatica underwent DCE MRI of the lumbar spine. After subtraction of dynamic images, a region of interest (ROI) was placed on each lumbar vertebral body of all subjects, and time intensity curves were generated. Consequently, perfusion parameters were calculated. Statistical analysis was performed to search for perfusion differences among lumbar vertebrae and in relation to age and sex. Upper (L1, L2) and lower (L3, L4, L5) vertebrae showed significant differences in perfusion parameters (p<0.05). Vertebrae of subjects younger than 50 years showed significantly higher perfusion compared to vertebrae of older ones (p<0.05). Vertebrae of females demonstrated significantly increased perfusion compared to those of males of corresponding age (p<0.05). All perfusion parameters, except for washout (WOUT), showed a mild linear correlation with age. Time to maximum slope (TMSP) and time to peak (TTPK) showed the same correlation with sex (0.227) in 68.3% of cases. A total of 75.9% respondents under-graded the TMA images. Gleason grading agreement with the expert reviewer correlated with the number of biopsies evaluated by the pathologist per week. Years of diagnostic experience, self description as a urologic pathologist or affiliation with a university hospital did not correlate with the pathologist's performance. CONCLUSION: The vast majority of participants under-graded the small tumors. Clinically relevant GS categories were correctly assigned in 68% of cases. This raises a potentially significant problem for pathologists, who have not had as much experience evaluating small PCas. PMID- 18392851 TI - Epidermal growth factor receptor gene mutation, amplification and protein expression in malignant pleural mesothelioma. AB - Epidermal growth factor receptor (EGFR) is overexpressed in a variety of epithelial malignancies. In lung cancer cases, EGFR gene mutation at the kinase domain and EGFR gene amplification are reported to be predictors of the response to EGFR tyrosine kinase inhibitors. In malignant pleural mesothelioma (MPM), the role of EGFR is less clear. We studied EGFR gene mutation, amplification and protein expression in 25 Japanese patients with MPM. None had previously reported EGFR mutations detected by the TaqMan PCR assay. Using immunohistochemistry, 8/25 (32%) cases were positive for the EGFR protein. The cases of sarcomatous type and desmoplastic type were all negative. Fluorescence in situ hybridization analysis revealed three low polysomy cases and one high polysomy case. The low polysomy cases included one biphasic type and two epithelial types, and the high polysomy case was epithelial type. These four cases expressed EGFR protein. In MPM, EGFR seems to play a role in a limited subset of patients. To identify possible candidates for EGFR tyrosine kinase in inhibitor therapy, the information on the EGFR gene status may be valuable. PMID- 18392852 TI - Prevalence of pre-malignant and malignant lesions in prophylactic mastectomy specimens of BRCA1 mutation carriers: comparison with a control group. AB - PURPOSE: BRCA1 mutation carriers are at high risk for breast cancer (BC). The risk management strategy may include radiological investigations for early detection or prophylactic mastectomy (PM). For a mutation carrier, PM may be more significant than surveillance alone when pre-malignant and malignant changes occur increasingly in mastectomy specimens, given normal findings on radiological investigations. In the present study we retrospectively investigated the differences between histological findings in PM specimens of BRCA1 carriers and those of a control group. METHODS: Twenty-four healthy and 28 affected carriers in the presence of normal preoperative radiological findings were included in the study. To compare the frequency of pre-malignant and malignant lesions in PM specimens, a control group matched for age and disease status was included. T tests for independent samples and Wilcoxon's signed-rank test were used for comparison of groups. RESULTS: The entire study group differed significantly from the control group (42.3 vs. 5.8%; P < 0.001) in terms of the occurrence of pre malignant and malignant lesions. Both, the sub-group comparison of healthy mutation carriers as well as diseased carriers with their controls, showed a significant difference in terms of the occurrence of pre-malignant and malignant changes (45.8 vs. 0%; P = 0.002; 39.3 vs. 10.7%; P = 0.03). In PM specimens of mutation carriers, carcinomas were identified in 5.8% (3/52) and pre-malignant changes in 36.5% (19/52). CONCLUSIONS: BRCA1 mutation carriers should be informed of the fact that pre-malignant and even malignant changes are frequently found in PM specimens despite normal radiological findings. PMID- 18392853 TI - Limited sequence variation in the major sperm protein 1 (MSP) gene within populations and species of the genus Dictyocaulus (Nematoda). AB - Populations of the bovine lungworm, Dictyocaulus viviparus, are genetically structured based on variation in mtDNA and AFLP data. Our aim was to investigate if this genetic variability also is reflected in a protein recognized by the host immune system. We focused on the major sperm protein (MSP), a small and abundant protein used in diagnostic immunoassays, which has been shown to be variable in some nematodes but not others. MSP was sequenced using worm DNA from eight adult worms from each of nine populations whose genetic structure previously had been quantified. For comparison, we also analyzed MSP sequences of the closely related Dictyocaulus eckerti and Dictyocaulus capreolus and from nematodes with sequences deposited in GenBank. In contrast to previous results, this study shows that the MSP ofD. viviparus is similar to that of other nematodes. Almost no sequence variation, and thus no antigenic diversity, was detected in MSP between worms from different sub-populations or in the other Dictyocaulus species investigated. A functional test of a recombinant variant of the MSP showed that the expressed protein was recognized by antibodies in sera from infected cattle. This has practical implications for the development of species-specific markers, recombinant vaccines, and immunodiagnostics. PMID- 18392854 TI - Characterization of chromosome ends on the basis of the structure of TrsA subtelomeric repeats in rice (Oryza sativa L.). AB - Subtelomeres contain species-specific repetitive sequences. We characterized rice chromosome ends on the basis of the structure of TrsA, a subtelomeric repetitive sequence of rice. Among the 24 chromosome arms, TrsA was arrayed in tandem on the ends of five: 5L, 6S, 8L, 9L, and 12L. TrsA sequences were arranged in discrete clusters of 3-106 copies in a chromosome-specific manner, instead of being distributed uniformly throughout the subtelomeric regions. The clusters were located at the distal-most end of the sequenced region in 5L, 6S, 8L, and 9L, but in 12L expressed genes were present distal to the clusters. Thus, rice subtelomeres are composed of discrete clusters of a TrsA-rich region and a gene rich region with high transcriptional activity. Intra-chromosomal duplications have resulted in a striking degree of variation in the number and distribution of TrsAs, suggesting that the areas near the ends of the chromosomes are dynamic and variable. PMID- 18392855 TI - Polymorphisms in 9q32 and TSCOT are linked to cervical cancer in affected sib pairs with high mean age at diagnosis. AB - Cervical cancer is a multifactorial disease influenced by both environmental and genetic factors. We have previously found linkage to 9q32 in a genomewide scan of affected sib-pairs (ASPs) with cervical cancer and to the thymic stromal co transporter (TSCOT), a candidate gene in this region. Here we examined the contribution of 9q32 and TSCOT to cervical cancer susceptibility using at larger material of 641 ASPs, 278 of which were included in the earlier genome-scan. Since heritable forms of cancer frequently show stronger genetic effects in families with early onset of disease, we stratified the ASPs into two groups based on mean age at diagnosis (MAAD) within sib-pairs. Surprisingly, ASPs with high MAAD (30.5-47.5 years) showed increased sharing at all microsatellite markers at 9q31.1-33.1 and linkage signals of up to MLS = 2.74 for TSCOT SNPs, while ASPs with low MAAD (19-30 years) showed no deviation from random genetic sharing (MLS = 0.00). The difference in allelic sharing between the two MAAD strata was significant (P < 0.005) and is not likely to be explained by the HLA haplotype, a previously known genetic susceptibility factor for cervical cancer. Our results indicate locus heterogeneity in the susceptibility to cervical cancer between the two strata, with polymorphisms in the 9q32 region mainly showing an effect in women with high MAAD. PMID- 18392856 TI - Hydrostatic constraints on morphological exploitation of light in tall Sequoia sempervirens trees. AB - We studied changes in morphological and physiological characteristics of leaves and shoots along a height gradient in Sequoia sempervirens, the tallest tree species on Earth, to investigate whether morphological and physiological acclimation to the vertical light gradient was constrained by hydrostatic limitation in the upper crown. Bulk leaf water potential (Psi) decreased linearly and light availability increased exponentially with increasing height in the crown. During the wet season, Psi was lower in the outer than inner crown. C isotope composition of leaves (delta(13)C) increased with increasing height indicating greater photosynthetic water use efficiency in the upper crown. Leaf and shoot morphology changed continuously with height. In contrast, their relationships with light availability were discontinuous: morphological characteristics did not correspond to increasing light availability above 55-85 m. Mass-based chlorophyll concentration (chl) decreased with increasing height and increasing light availability. In contrast, area-based chl remained constant or increased with increasing height. Mass-based maximum rate of net photosynthesis (P (max)) decreased with increasing height, whereas area-based P (max) reached maximum at 78.4 m and decreased with increasing height thereafter. Mass-based P (max) increased with increasing shoot mass per area (SMA), whereas area-based P (max) was not correlated with SMA in the upper crown. Our results suggest that hydrostatic limitation of morphological development constrains exploitation of light in the upper crown and contributes to reduced photosynthetic rates and, ultimately, reduced height growth at the tops of tall S. sempervirens trees. PMID- 18392857 TI - Controls over nitric oxide and ammonia emissions from Mojave Desert soils. AB - Emissions of reactive N compounds produced during terrestrial N cycling can be an important N loss pathway from ecosystems. Most measurements of this process focus on NO and N(2)O efflux; however, in alkaline soils such as those in the Mojave Desert, NH(3) production can be an important component of N gas loss. We investigated patterns of NO and NH(3) emissions in the Mojave Desert and identified seasonal changes in temperature, precipitation and spatial heterogeneity in soil nutrients as primary controllers of soil efflux. Across all seasons, NH(3) dominated reactive N gas emissions with fluxes ranging from 0.9 to 10 ng N m(-2) s(-1) as compared to NO fluxes of 0.08-1.9 ng N m(-2) s(-1). Fluxes were higher in April and July than in October; however, a fall precipitation event yielded large increases in both NO and NH(3) efflux. To explore the mechanisms driving field observations, we combined NO and NH(3) soil flux measurements with laboratory manipulations of temperature, water and nutrient conditions. These experiments showed a large transient NH(3) pulse (~70-100 ng N m(-2) s(-1)) following water addition, presumably driven by an increase in soil NH(4) (+) concentrations. This was followed by an increase in NO production, with maximum NO flux rates of 34 ng N m(-2) s(-1). Our study suggests that immediately following water addition NH(3) volatilization proceeds at high rates due to the absence of microbial competition for NH(4) (+); during this period N gas loss is insensitive to changes in temperature and soil nutrients. Subsequently, NO emission increases and rates of both NO and NH(3) emission are sensitive to temperature and nutrient constraints on microbial activity. Addition of labile C reduces gaseous N losses, presumably by increasing microbial immobilization, whereas addition of NO(3) (-) stimulates NO and NH(3) efflux. PMID- 18392858 TI - Herbivore benefits from vectoring plant virus through reduction of period of vulnerability to predation. AB - Herbivores can profit from vectoring plant pathogens because the induced defence of plants against pathogens sometimes interferes with the induced defence of plants against herbivores. Plants can also defend themselves indirectly by the action of the natural enemies of the herbivores. It is unknown whether the defence against pathogens induced in the plant also interferes with the indirect defence against herbivores mediated via the third trophic level. We previously showed that infection of plants with Tomato spotted wilt virus (TSWV) increased the developmental rate of and juvenile survival of its vector, the thrips Frankliniella occidentalis. Here, we present the results of a study on the effects of TSWV infections of plants on the effectiveness of three species of natural enemies of F. occidentalis: the predatory mites Neoseiulus cucumeris and Iphiseius degenerans, and the predatory bug Orius laevigatus. The growth rate of thrips larvae was positively affected by the presence of virus in the host plant. Because large larvae are invulnerable to predation by the two species of predatory mites, this resulted in a shorter period of vulnerability to predation for thrips that developed on plants with virus than thrips developing on uninfected plants (4.4 vs. 7.9 days, respectively). Because large thrips larvae are not invulnerable to predation by the predatory bug Orius laevigatus, infection of the plant did not affect the predation risk of thrips larvae from this predator. This is the first demonstration of a negative effect of a plant pathogen on the predation risk of its vector. PMID- 18392859 TI - Population variation in plant traits associated with ant attraction and herbivory in Chamaecrista fasciculata (Fabaceae). AB - The benefits of ant-plant-herbivore interactions for the plant depend on the abundance of ants and herbivores and the selective pressures these arthropods exert. In plants bearing extrafloral nectaries (EFN), different mean trait values may be selected for by different populations in response to local herbivore pressure, ultimately resulting in the evolution of differences in plant traits that attract ants as defensive agents against herbivory. To determine if variation in traits that mediate ant-plant interactions reflect herbivore selective pressures, we quantified intra- and inter-population variation in plant traits for eight populations of the EFN-bearing annual Chamaecrista fasciculata (Michx.) (Fabaceae). Censuses in rural and urban areas of Missouri and Illinois (USA) showed population differences in ant attendance and herbivore pressure. Seeds were collected from each population, and plants were grown in a common greenhouse environment to measure sugar production, nectar volume and composition, EFN size and time of emergence, leaf pubescence, and leaf quality throughout plant development. Populations varied mainly in terms of nectary size, sugar production, and nectar volume, but to a lesser degree in leaf pubescence. Populations of C. fasciculata within urban areas (low in insect abundance) had small nectaries and the lowest nectar production. There was a positive correlation across populations between herbivore density and leaf damage by those herbivores on the one hand and sugar production and nectar volume on the other. These results, in conjunction with lack of evidence for maternally based environmental effects, suggest that population differences in herbivore damage have promoted differential evolution of EFN-related traits among populations. PMID- 18392860 TI - Hemodialysis vascular access options in pediatrics: considerations for patients and practitioners. AB - Recent data indicate that the incidence of end-stage renal disease (ESRD) in pediatric patients (age 0-19 years) has increased over the past two decades. Similarly, the prevalence of ESRD has increased threefold over the same period. Hemodialysis (HD) continues to be the most frequently utilized modality for renal replacement therapy in incident pediatric ESRD patients. The number of children on HD exceeded the sum total of those on peritoneal dialysis and those undergoing pre-emptive renal transplantation. Choosing the best vascular access option for pediatric HD patients remains challenging. Despite a national initiative for fistula first in the adult hemodialysis population, the pediatric nephrology community in the United States of America utilizes central venous catheters as the primary dialysis access for most patients. Vascular access management requires proper advance planning to assure that the best permanent access is placed, seamless communication involving a multidisciplinary team of nephrologists, nurses, surgeons, and interventional radiologists, and ongoing monitoring to ensure a long life of use. It is imperative that practitioners have a long-term vision to decrease morbidity in this unique patient population. This article reviews the various types of pediatric vascular accesses used worldwide and the benefits and disadvantages of these various forms of access. PMID- 18392861 TI - General perceived self-efficacy: validation analysis in Greek cancer patients. AB - GOALS OF WORK: The present study aims to validate the General Perceived Self efficacy Scale (GSE) in Greek cancer patients. MATERIALS AND METHODS: The scale was administered twice, with a 3-day interval, to 99 advanced cancer patients. The patients also completed the Anxiety Subscale from the Greek Hospital Depression and Anxiety Scale. MAIN RESULTS: Factor analyses identified a one factor solution, explaining 74.6% of the variance. The Greek version of the GSE had a Cronbach's alpha of 0.927. Validity as performed using known-group analysis showed good results. Satisfactory construct validity was supported by the correlation analysis between the GSE and anxiety (r = -0.507, p < 0.0005). Interitem correlations was also satisfactory at p < 0.0005. CONCLUSIONS: These psychometric properties of the Greek version of the GSE showed that it is a valid and reliable measured when administered to cancer patients. PMID- 18392862 TI - Impact of skeletal complications on patients' quality of life, mobility, and functional independence. AB - INTRODUCTION: Skeletal-related events (SREs) from malignant bone disease cause considerable morbidity and can dramatically reduce patients' quality of life. DISCUSSION: Pathologic fractures often require surgical intervention and palliative radiotherapy. Thus, patients suffer impaired mobility, loss of functional independence, and diminished health-related quality of life (HRQOL). Bisphosphonates can delay the onset and reduce the incidence of SREs and have become the standard of care for the treatment of malignant bone disease; however, minimal information on the effects of bisphosphonate treatment on HRQOL is available. Targeted HRQOL assessments for patients with malignant bone disease are currently under development and are discussed herein. PMID- 18392863 TI - Submolecular unfolding units of Pseudomonas aeruginosa cytochrome c-551. AB - Hydrogen exchange rates for backbone amide protons of oxidized Pseudomonas aeruginosa cytochrome c-551 (P. aeruginosa cytochrome c) have been measured in the presence of low concentrations of the denaturant guanidine hydrochloride. Analysis of the data has allowed identification of submolecular unfolding units known as foldons. The highest-energy foldon bears similarity to the proposed folding intermediate for P. aeruginosa cytochrome c. Parallels are seen to the foldons of the structurally homologous horse cytochrome c, although the heme axial methionine-bearing loop has greater local stability in P. aeruginosa cytochrome c, in accord with previous folding studies. Regions of low local stability are observed to correspond with regions that interact with redox partners, providing a link between foldon properties and function. PMID- 18392864 TI - The effect of mutation of F87 on the properties of CYP102A1-CYP4C7 chimeras: altered regiospecificity and substrate selectivity. AB - CYP102A1 is a highly active water-soluble bacterial monooxygenase that contains both substrate-binding heme and diflavin reductase subunits, all in a single polypeptide that has been called a "self-sufficient enzyme." Several years ago we developed a procedure called "scanning chimeragenesis," where we focused on residues 73-82 of CYP102A1, which contact approximately 40% of the substrates palmitoleic acid and N-palmitoylglycine [Murataliev et al. (2004) Biochemistry 43:1771-1780]. These residues were replaced with the homologous residues of CYP4C7. In the current work, that study has been expanded to include residue 87. Phenylalanine 87 of wild-type CYP102A1 was replaced with the homologous residue of CYP4C7, leucine, as well as with alanine. The full-sized chimeric proteins C(73-78, F87L), C(73-78, F87A), C(75-80, F87L), C(75-80, F87A), C(78-82, F87L) and C(78-82, F87A) have been purified and characterized. Wild-type CYP102A1 is most active toward fatty acids (both lauric and palmitic acids produce omega-1, omega-2, and omega-3 hydroxylated fatty acids), but it also catalyzes the oxidation of farnesol to three products (2, 3- and 10,11-epoxyfarnesols and 9 hydroxyfarnesol). All of the F87-mutant chimeric proteins show dramatic decreases in activities with the natural CYP102A1 substrates. In contrast, C(78-82, F87A) and C(78-82, F87L) have markedly increased activities with farnesol, with the latter showing a 5.7-fold increase in catalytic activity as compared to wild-type CYP102A1. C(78-82, F87L) produces 10,11-epoxyfarnesol as the single primary metabolite. The results show that chimeragenesis involving only the second half of SRS-1 plus F87 is sufficient to change the substrate selectivity of CYP102A1 from fatty acids to farnesol and to produce a single primary product. PMID- 18392865 TI - Progranulin genetic variations in frontotemporal lobar degeneration: evidence for low mutation frequency in an Italian clinical series. AB - Frontotemporal lobar degeneration (FTLD) recognises high familial incidence, with up to 50% of patients reported to have a family history of similar dementia. It has been reported that mutations within progranulin (PGRN) gene are a major cause of FTLD in the USA and worldwide, counting for 5-10% of FTLD and for 20-25% of familiar FTLD cases. The aim of the present study was to define the role of PGRN genetic variations in a large sample of consecutive patients with FTLD in Italy. Two-hundred forty-three FTLD patients were investigated. Each subject performed a clinical and neuropsychological evaluation, a functional and structural brain imaging, and the diagnosis was confirmed by at least 1 year follow-up. PGRN sequencing was performed in all FTLD patients and in 121 healthy age-matched controls drawn from the same geographic area. Only one PGRN pathogenetic mutation was found, consisting of a four-base pair deletion in the coding sequence of exon 8 (delCACT). This mutation was recognised in four patients, being the overall frequency of mutations in our clinical series of 1.64%. Considering only patients with a well-known family history for dementia, the frequency of this mutation was 6%. Moreover, four missense mutations within intron regions (g.100474G>A, g.100674G>A, g.101266G>A, g.102070G>A) were found. The frequency of these genetic variations did not differ in patients compared to controls, and they did not influence on clinical FTLD phenotype. In conclusion, this study supports a lower frequency of PGRN mutations amongst FTLD patients in Italy compared to literature data and further underlies the genetic heterogeneity of FTLD. PMID- 18392866 TI - Limited efficacy of a nonrestricted intervention on antimicrobial prescription of commonly used antibiotics in the hospital setting: results of a randomized controlled trial. AB - Most interventions aimed at diminishing the use of antimicrobials in hospitals have focussed on newly introduced antibiotics and very few have been randomly controlled. We evaluated the impact on antibiotic consumption of an intervention without restrictions in antibiotic use, focussed on commonly used antibiotics with a controlled randomized trial. All new prescriptions of levofloxacin, carbapenems, or vancomycin in hospitalized patients were randomized to an intervention or a control group. Intervention consisted of an antibiotic regimen counselling targeted to match local antibiotic guidelines, performed using only patients' charts. Clinical charts of patients assigned to the control group were reviewed daily by a pharmacist. The primary endpoint was a reduction in consumption of the targeted antibiotics. Two hundred seventy-eight prescriptions corresponding to 253 patients were included: 146 were assigned to the intervention and 132 to the control group. Total consumption of the targeted antibiotics (median [IQR]) was slightly lower in the intervention (8 [4-12] defined daily doses [DDDs] per patient) than in the control group (10 [6-16] DDDs per patient; p = 0.04). No differences in number of DDDs were observed when antibiotics of substitution were included (11.05 [6-18.2] vs 10 [6-16.5] in the intervention and control groups, respectively, p = 0.13). The total number of days on treatment with the targeted antibiotics was lower in the intervention (4 [3-7] days per patient) than in the control group (6 [4-10] days per patient; p = 0.002). Differences in number of days on treatment only reached statistical significance in the prescriptions of carbapenems. There were no differences between intervention and control groups in terms of number of deaths, hospital readmissions, length of hospital stay, or antibiotic costs. In this trial, an intervention without restrictions focussed on antimicrobial prescriptions of commonly used antibiotics in the hospital setting had a limited efficacy to reduce consumption and did not save costs. Future strategies to promote a more rational antimicrobial use should be evaluated with a randomized controlled design. PMID- 18392867 TI - The relationship between the Balanced Budget Act and length of stay for Medicare patients in US hospitals. AB - BACKGROUND: The Balanced Budget Act (BBA) of 1997 and Balanced Budget Refinement Act (BBRA) of 1999 led to deep financial cuts for hospitals and nursing homes. OBJECTIVES: We examine the effects of these acts on hospital length of stay (LOS) for Medicare recipients. METHODS: Using data for all short-stay community hospitals in the country, we compared LOS for Medicare patients before and after the BBA/BBRA relative to known determinants of LOS, e.g., hospital ownership, region, beds, financial performance, and conversion/change in ownership type. RESULTS: Hospital LOS was reduced as a result of the acts. Reductions were more apparent for larger urban hospitals that provided safety-net services. LOS varied slightly by hospital ownership. CONCLUSION: This study is among the first to evaluate the impact of BBA and BBRA on hospital services. These acts had a negative effect on the ability of hospitals to continue offering safety-net services and negatively affected LOS. PMID- 18392868 TI - Chemostat selection of a bacterial community able to degrade s-triazinic compounds: continuous simazine biodegradation in a multi-stage packed bed biofilm reactor. AB - Using a successive transfer method on mineral salt medium containing simazine, a microbial community enriched with microorganisms able to grow on simazine was obtained. Afterwards, using a continuous enrichment culture procedure, a bacterial community able to degrade simazine from an herbicide formulation was isolated from a chemostat. The continuous selector, fed with a mineral salt medium containing simazine and adjuvants present in the commercial herbicide formulation, was maintained in operation for 42 days. Following the lapse of this time, the cell count increased from 5 x 10(5) to 3 x 10(8) CFU mL(-1), and the simazine removal efficiency reached 96%. The chemostat's bacterial diversity was periodically evaluated by extracting the culture's bacterial DNA, amplifying their 16S rDNA fragments and analyzing them by thermal gradient gel electrophoresis. Finally, a stable bacterial consortium able to degrade simazine was selected. By PCR amplification, sequencing of bacterial 16S rDNA amplicons, and comparison with known sequences of 16S rDNA from the NCBI GenBank, eight bacterial strains were identified. The genera, Ochrobactrum, Mycobacterium, Cellulomonas, Arthrobacter, Microbacterium, Rhizobium and Pseudomonas have been reported as common degraders of triazinic herbicides. On the contrary, we were unable to find reports about the ability of the genus Pseudonocardia to degrade triazinic compounds. The selected bacterial community was attached to a porous support in a concurrently aerated four-stage packed-bed reactor fed with the herbicide. Highest overall simazine removal efficiencies eta (SZ) were obtained at overall dilution rates D below 0.284 h(-1). However, the multistage packed bed reactor could be operated at dilution rates as high as D = 3.58 h(-1) with overall simazine removal volumetric rates R (v,SZ) = 19.6 mg L(-1) h(-1), and overall simazine removal specific rates R (X,SZ) = 13.48 mg (mg cell protein)(-1) h(-1). Finally, the consortium's ability to degrade 2-chloro-4,6-diamino-1,3,5 triazine (CAAT), cyanuric acid and the herbicide atrazine, pure or mixed with simazine, was evaluated in fed batch processes. PMID- 18392869 TI - Biomass measurement online: the performance of in situ measurements and software sensors. AB - Biomass measurement is one of the most critical measurements in biotechnological processes. The technologies developed for the measurement of biomass in situ have developed over the years. Because it has been over 10 years since the last review concentrating on practical issues concerning biomass measurements, it is time to evaluate recent developments in the field. This review concentrates on the applications of dielectric spectroscopy, optical density, infrared spectroscopy, and fluorescence for in situ measurement of biomass. The advantages offered by these methods and an economic way of estimating biomass concentration, the software sensors, are considered. PMID- 18392870 TI - Whole cell microbial transformation in cloud point system. AB - Cloud point system, consisting of nonionic surfactant in an aqueous solution, has been developed as a novel medium for whole cell microbial transformation. The basic properties of cloud point system including phase separation and solubilization are introduced. The application of cloud point system for extractive microbial transformation is different from that of water-organic solvent two-phase partitioning system or aqueous two-phase system are discussed, which mainly focus on the biocompatibility of microorganism in a cloud point system and a downstream process of microbial transformation in cloud point system with oil-water-surfactant microemulsion liquid-liquid extraction for surfactant recovery and product separation. Finally, examples of whole cell microbial transformation in cloud point systems, especially in situ extraction of moderate polar substrate/product, are also presented. PMID- 18392871 TI - Analysis of multiple waveforms by means of functional principal component analysis: normal versus pathological patterns in sit-to-stand movement. AB - This paper presents an application of functional principal component analysis (FPCA) to describe the inter-subject variability of multiple waveforms. This technique was applied to the study of sit-to-stand movement in two groups of people, osteoarthritic patients and healthy subjects. Although STS movement has not been extensively applied to the study of knee osteoarthritis, it can provide relevant information about the effect of osteoarthritis on knee joint function. Two waveforms, knee flexion angle and flexion moment, were analysed simultaneously. Instead of using the common multivariate approach we used the functional one, which allows working with continuous functions with neither discretization nor time-scale normalization. The results show that time-scale normalization can alter the FPCA solution. Furthermore, FPCA presents better discriminatory power compared with the classical multivariate approach. This technique can, therefore, be applied as a functional assessment tool, allowing the identification of relevant variables to discriminate heterogeneous groups such as healthy and pathological subjects. PMID- 18392872 TI - N-3 polyunsaturated fatty acids modulate in-vitro T cell function in type I diabetic patients. AB - In this work, we assessed the in-vitro effects of eicosapentaenoic acid (EPA; C20:5n-3) and docosahexaenoic acid (DHA; C22:6n-3) (final concentration, 15 microM) on T cell blastogenesis, interleukin-2 and -4 (IL-2, IL-4) secretion, fatty acid composition and intracellular oxidative status in type I diabetic patients with or without complications. Con A stimulated lymphocyte proliferation, glucose uptake, intracellular reduced glutathione levels and catalase activity were lower in diabetics as compared to controls, regardless to the presence of complications. EPA and DHA diminished T-lymphocyte proliferation and IL-2 production but enhanced IL-4 secretion in both diabetic and control groups. No changes in the levels of reduced glutathione and in the activities of catalase and SOD were observed in control T cells cultured in the presence of EPA and DHA. However, in diabetic patients, addition of n-3 PUFA to culture induced an increase in T cell levels of reduced glutathione and hydroperoxide, and in activities of catalase and SOD. Low levels of arachidonic acid (C20:4n-6) were found in plasma membrane phospholipids of lymphocytes from diabetic patients compared to controls. Incubation of lymphocytes with EPA and DHA was associated with an incorporation of these fatty acids in membrane phospholipids. In conclusion, the beneficial effects of n-3 PUFA on T cell functions in type I diabetes could be attributed to their suppressive action and modulation of cytokine secretion, and to the improvement of intracellular oxidative status. PMID- 18392873 TI - Development of Tribolium castaneum. PMID- 18392874 TI - From development to biodiversity--Tribolium castaneum, an insect model organism for short germband development. AB - Insect embryogenesis is best understood in the fruit fly Drosophila. However, Drosophila embryogenesis shows evolutionary-derived features: anterior patterning is controlled by a highly derived Hox gene bicoid, the body segments form almost simultaneously and appendages develop from imaginal discs. In contrast, embryogenesis of the red flour beetle Tribolium castaneum displays typical features in anterior patterning, axis and limb formation shared with most insects, other arthropods as well as with vertebrates. Anterior patterning depends on the conserved homeobox gene orthodenticle, the main body axis elongates sequentially and limbs grow continuously starting from an appendage bud. Thus, by analysing developmental processes in the beetle at the molecular and cellular level, inferences can be made for similar processes in other arthropods. With the completion of sequencing the Tribolium genome, the door is now open for post-genomic studies such as RNA expression profiling, proteomics and functional genomics to identify beetle-specific gene circuits. PMID- 18392875 TI - Analysis of the Tribolium homeotic complex: insights into mechanisms constraining insect Hox clusters. AB - The remarkable conservation of Hox clusters is an accepted but little understood principle of biology. Some organizational constraints have been identified for vertebrate Hox clusters, but most of these are thought to be recent innovations that may not apply to other organisms. Ironically, many model organisms have disrupted Hox clusters and may not be well-suited for studies of structural constraints. In contrast, the red flour beetle, Tribolium castaneum, which has a long history in Hox gene research, is thought to have a more ancestral-type Hox cluster organization. Here, we demonstrate that the Tribolium homeotic complex (HOMC) is indeed intact, with the individual Hox genes in the expected colinear arrangement and transcribed from the same strand. There is no evidence that the cluster has been invaded by non-Hox protein-coding genes, although expressed sequence tag and genome tiling data suggest that noncoding transcripts are prevalent. Finally, our analysis of several mutations affecting the Tribolium HOMC suggests that intermingling of enhancer elements with neighboring transcription units may constrain the structure of at least one region of the Tribolium cluster. This work lays a foundation for future studies of the Tribolium HOMC that may provide insights into the reasons for Hox cluster conservation. PMID- 18392876 TI - Do teashirt family genes specify trunk identity? Insights from the single tiptop/teashirt homolog of Tribolium castaneum. AB - The Drosophila teashirt gene acts in concert with the homeotic selector (Hox) genes to specify trunk (thorax and abdomen) identity. There has been speculation that this trunk-specifying function might be very ancient, dating back to the common ancestor of insects and vertebrates. However, other evidence suggests that the role of teashirt in trunk identity is not well conserved even within the Insecta. To address this issue, we have analyzed the function of Tc-tiotsh, the lone teashirt family member in the red flour beetle, Tribolium castaneum. Although Tc-tiotsh is important for aspects of both embryonic and imaginal development including some trunk features, we find no evidence that it acts as a trunk identity gene. We discuss this finding in the context of recent insights into the evolution and function of the Drosophila teashirt family genes. PMID- 18392877 TI - Sites of Fgf signalling and perception during embryogenesis of the beetle Tribolium castaneum. AB - The development of multicellular embryos depends on coordinated cell-to-cell signalling events. Among the numerous cell-signalling pathways, fibroblast growth factors (FGFs) are involved in important processes during embryogenesis, such as mesoderm formation during gastrulation and growth. In vertebrates, the Fgf superfamily consists of 22 family members, whereas only few FGFs are contained in the less complex genomes of insects and worms. In the recently sequenced genome of the beetle Tribolium, we identified four Fgf family members representing three subfamilies. Tribolium has Fgf1 genes that are absent in Drosophila but known from vertebrates. By phylogenetic analysis and microsynteny to Drosophila, we further classify Tc-fgf 8 as an ancestor of pyramus and thisbe, the fly Fgf8 genes. Tc-fgf8 expression in the growth zone suggests an involvement in mesoderm formation. In the embryonic head, expression of Tc-fgf8 subdivides the brain into a larger anterior and a smaller posterior region. The Fgf Tc-branchless is expressed in the embryonic tracheal placodes and in various gland-like structures. The expression patterns of the only Tribolium Fgf receptor and the adaptor molecule Downstream-of-Fgfr are largely congruent with Tc-Fgf8 and Tc bnl. Thus, in contrast to Drosophila, only one Fgf receptor canalises Fgf signalling in different tissues in Tribolium. Our findings significantly advance our understanding of the evolution of Fgf signalling in insects. PMID- 18392878 TI - Expression, function and regulation of Brachyenteron in the short germband insect Tribolium castaneum. AB - T-domain transcription factors are involved in many different processes during embryogenesis, such as mesoderm, heart or gut development in vertebrates and in invertebrates. In insects, the following five types of T-box genes are known: brachyenteron (byn), optomotor-blind (omb), optomotor-blind-related-gene-1 (org 1), dorsocross (doc) and H15. As all these classes are present in the genome of the fruit fly Drosophila melanogaster and the flour beetle Tribolium, the multiplicity of the five types of genes varies from dipterans to the beetle. In higher dipterans, a small cluster of three doc genes (doc1-doc3) exists, while the Tribolium genome contains a single Tc-doc gene only. Two H15 genes, Tc-H15a and Tc-H15b, are present in the Tribolium genome compared to a single H15 gene in Drosophila. We have analysed the expression and function of the Tribolium brachyenteron ortholog (Tc-byn). During embryogenesis, Tc-byn is exclusively expressed in the growth zone of the extending germband and later becomes confined to the distal proctodeum and the hindgut, a situation that parallels the expression pattern of byn in Drosophila. Tc-byn-RNAi treated embryos phenocopy Drosophila byn mutants and form no hindgut. In addition, we have characterised a regulatory element upstream of the Tc-byn transcription start site that confers specific gene expression in the developing hindgut of the Drosophila embryo. Our results demonstrate a highly conserved role for Brachyury-type transcriptional regulators in posterior gut development of insects at the level of expression, function and regulation. PMID- 18392879 TI - Hedgehog signaling pathway function conserved in Tribolium segmentation. AB - In Drosophila, maintenance of parasegmental boundaries and formation of segmental grooves depend on interactions between segment polarity genes. Wingless and Engrailed appear to have similar roles in both short and long germ segmentation, but relatively little is known about the extent to which Hedgehog signaling is conserved. In a companion study to the Tribolium genome project, we analyzed the expression and function of hedgehog, smoothened, patched, and cubitus interruptus orthologs during segmentation in Tribolium. Their expression was largely conserved between Drosophila and Tribolium. Parental RNAi analysis of positive regulators of the pathway (Tc-hh, Tc-smo, or Tc-ci) resulted in small spherical cuticles with little or no evidence of segmental grooves. Segmental Engrailed expression in these embryos was initiated but not maintained. Wingless independent Engrailed expression in the CNS was maintained and became highly compacted during germ band retraction, providing evidence that derivatives from every segment were present in these small spherical embryos. On the other hand, RNAi analysis of a negative regulator (Tc-ptc) resulted in embryos with ectopic segmental grooves visible during germband elongation but not discernible in the first instar larval cuticles. These transient grooves formed adjacent to Engrailed expressing cells that encircled wider than normal wg domains in the Tc ptc RNAi embryos. These results suggest that the en-wg-hh gene circuit is functionally conserved in the maintenance of segmental boundaries during germ band retraction and groove formation in Tribolium and that the segment polarity genes form a robust genetic regulatory module in the segmentation of this short germ insect. PMID- 18392880 TI - Tribolium Wnts: evidence for a larger repertoire in insects with overlapping expression patterns that suggest multiple redundant functions in embryogenesis. AB - Wingless (wg)/Wnt family genes encode secreted glycoproteins that function as signalling molecules in the development of vertebrates as well as invertebrates. In a survey of Wnt family genes in the newly sequenced Tribolium genome, we found a total of nine Wnt genes. In addition to wg or Wnt1, Tribolium contains orthologs of the vertebrate Wnt5-7 and Wnt9-11 genes. As in Drosophila, Wnt1, Wnt6 and Wnt10 are clustered in the genome. Comparative genomics indicates that Wnt9 is also a conserved member of this cluster in several insects for which genome sequence is available. One of the Tribolium Wnt genes appears to be a member of the WntA family, members of which have been identified in Anopheles and other invertebrates but not in Drosophila or vertebrates. Careful phylogenetic examination suggests an Apis Wnt gene, previously identified as a Wnt4 homolog, is also a member of the WntA family. The ninth Tribolium Wnt gene is related to the diverged Drosophila WntD gene, both of which phylogenetically group with Wnt8 genes. Some of the Tribolium Wnt genes display multiple overlapping expression patterns, suggesting that they may be functionally redundant in segmentation, brain, appendage and hindgut development. In contrast, the unique expression patterns of Wnt5, Wnt7 and Wnt11 in developing appendages likely indicate novel functions. PMID- 18392881 TI - TGFbeta signaling in Tribolium: vertebrate-like components in a beetle. AB - The cytokines of the TGFbeta superfamily are highly conserved in evolution and elicit a diverse range of cellular responses in all metazoa. In Drosophila, the signaling pathways of the two TGFbeta subfamilies, Activins and Bone Morphogenetic Proteins (BMPs), have been well studied. To address the question of whether the findings from Drosophila are representative of insects in general, we analyzed the components of TGFbeta-signaling present in the genome of the beetle Tribolium castaneum. We were able to identify orthologs of the BMPs Decapentaplegic and Glass bottom boat, of the Activins Activinbeta and Dawdle, as well as orthologs of the less well-known ligands Myoglianin and Maverick, together with orthologs of all TGFbeta receptors and cytoplasmic signal transducers present in Drosophila. This indicates that the diversity of TGFbeta signaling components is generally well conserved between Drosophila and Tribolium. However, the genome of the beetle-and of the bee Apis mellifera-lacks an ortholog of the Drosophila BMP Screw but does contain a vertebrate-like BMP10 homolog which is not found in Drosophila. Concerning BMP inhibitors, Tribolium displays an even more vertebrate-like ensemble of components. We found two orthologs of the vertebrate DAN family, Dan and Gremlin, and show embryonic expression of a vertebrate-like BAMBI ortholog, all of which are absent in Drosophila. This suggests that Tribolium might have retained a more ancestral composition of TGFbeta signaling components and that TGFbeta signaling underwent considerable change in the Drosophila lineage. Tribolium is an excellent model to study the function of these ancestral signaling components in insects. PMID- 18392882 TI - Fast co-evolution of sevenless and bride of sevenless in endopterygote insects. AB - Activation of Ras signaling by the receptor tyrosine kinase Sevenless plays important roles during retinal patterning and male germline development in Drosophila. Sevenless is orthologous to the vertebrate receptor tyrosine kinase c ros. Remarkably, vertebrate ligands of c-Ros as well as non-Drosophila orthologs of the Sevenless ligand Bride of sevenless have remained elusive. Using newly available insect genome sequence information, we investigated the evolutionary conservation of the seven transmembrane domain protein gene bride of sevenless. Single orthologs were identified in the genomes of mosquito, flour beetle, and honeybee due to strong sequence conservation in the seven transmembrane domain. The extracellular region, however, is only detectably conserved within but not outside Diptera. Analysis of domain-specific substitution rates demonstrates correlated fast rates of evolutionary change in the extracellular domains of both bride of sevenless and sevenless. The rapid pace of sequence change explains why Sevenless ligands are difficult to detect by sequence similarity in distantly related phyla. Second, the conservation of bride of sevenless in flour beetle and honeybee raises the possibility of conserved Sevenless signaling controlled patterning processes in endopterygote insects. PMID- 18392883 TI - Aquatic toxicity assessment of the additive 6-methylcoumarine using four experimental systems. AB - The toxicity assessment of chemicals is one of the main issues in the current policies in order to protect the health of the environment and human beings. Food and cosmetic additives have been extensively studied in relation to their toxicity to humans, but data about their ecotoxicological effects are scarce. The aim of this study was to evaluate the toxic effects of the additive 6 methylcoumarine in the aquatic milieu using a test battery comprising experimental model systems from different trophic levels. The inhibition of bioluminiscence was studied in the bacteria Vibrio fischeri (decomposer), the inhibition of growth was evaluated in the alga Chlorella vulgaris (producer) and immobilization was studied in the cladoceran Daphnia magna (first consumer). Finally, several end points were evaluated in the RTG-2 salmonid fish cell line, including neutral red uptake, protein content, methylthiazol tetrazolium salt metabolization, glucose-6-phosphate dehydrogenase activity, lactate dehydrogenase activity and leakage, and morphology. The sensitivity of the test systems employed was as follows: V. fischeri > D. magna > C. vulgaris > RTG-2 cell line. The results show that 6-methylcoumarine is not expected to produce acute toxic effects on the aquatic biota. However, chronic and synergistic effects with other chemicals cannot be excluded and should be further investigated. PMID- 18392884 TI - Postoperative management of ala nasi surgery: a new device. PMID- 18392885 TI - Filling of adjustable breast implants beyond the manufacturer's recommended fill volume. AB - BACKGROUND: Adjustable breast implants are widely used for both reconstructive and cosmetic breast surgery. They provide unique postoperative versatility and allow for more effective management of numerous conditions that would otherwise require surgical intervention. Findings have shown that in a clinical setting, it often is necessary to overexpand saline implants beyond the manufacturer's recommended fill volumes for positive results and optimal patient satisfaction. The authors investigated their breast-augmentation patients, comparing implants expanded beyond the manufacturer's fill volume with implants that remained within the recommended parameters. METHODS: A total of 138 patients (270 implants) undergoing breast augmentation mammoplasty with Smooth Round Spectrum implants were evaluated postoperatively. To determine the effects of overexpansion, the incidence of leakage was assessed as well as the possibility of increased firmness. Patients completed a satisfaction questionnaire. RESULTS: The findings showed no evidence of increased leakage with implants expanded beyond recommended fill volumes, and 97% of the patients reported a perfect score for satisfaction ratings. Of the 270 implants evaluated in the study, only 7 were given a less than perfect score. CONCLUSIONS: By exceeding the recommended fill volume, the authors were able to use the implant to its maximal potential, with increased patient satisfaction. The authors were able to correct problems and avoid complications that could not have been resolved without deviating from the manufacturer's recommended fill volumes. Nonvalidated restrictions on fill volumes severely limit the efficacy of adjustable implants. PMID- 18392886 TI - Microvascular transplantation of adipose tissue and serum level of adipocyte products. AB - Microvascular transplantation of subcutaneous adipose tissue is an essential step in reconstructive surgery after breast carcinoma. Serum levels of adipose tissue products may serve as indicators for transplant function. This study aimed to determine serum leptin and tumor necrosis factor (TNF)-alpha plasma levels pre-, intra-, and postoperatively in 20 patients undergoing reconstructive breast surgery and in 7 women undergoing abdominoplasty operation. In the patients undergoing reconstructive breast surgery, the serum leptin levels decreased intraoperatively from 14.5 +/- 13.1 to 9.1 +/- 7.3 ng/ml, a decrease of 63%. An increase in serum leptin levels to 13.5 +/- 12.7 ng/ml (93% of the initial value) was found on postoperative day 1. This was paralleled by similar changes in the plasma levels of TNF-alpha (preoperatively, 20 +/- 7.3 pg/ml; intraoperatively, 17 +/- 11.4 pg/ml; postoperatively, 21 +/- 10.8 pg/ml). In the patients undergoing abdominoplasty, plasma leptin and TNF-alpha levels decreased intraoperatively (20% and 27%, respectively) and postoperatively (44% and 27%, respectively). The results of our pilot study indicate that a postoperative increase in the level of serum leptin after reconstructive breast surgery may be related to successful transplant function. PMID- 18392887 TI - Private sector participation in urban water and sanitation provision in Ghana: experiences from the Tamale Metropolitan Area (TMA). AB - African governments, like most countries in the developing world, face daunting tasks in their attempts to provide effective and equitable water and sanitation services for their ever increasing urban populations. Consequently, the past few years have witnessed increased private sector participation in urban water and sanitation provision, as many African governments strive to improve access to water and sanitation services for their citizens in line with Millennium Development Goal 7 (MDG7). Since the early 1990s, the government of Ghana and many local authorities have entered into various forms of public-private partnerships in urban water and sanitation provision. This article examines the outcome of such partnerships using the Tamale Metropolitan Area (TMA) as a case study with the aim of providing policy guidelines for the way forward. The article argues that the public-private arrangement for water supply and sanitation infrastructure management in the Tamale Metropolis has done nothing that an invigorated public sector could not have possibly achieved. It concludes that there can be no sustainable improvement in water and sanitation provision without political commitment, stakeholder ownership, and strong support for community driven initiatives. PMID- 18392888 TI - Land degradation on barren hills: a case study in northeast Vietnam. AB - The term "barren hills" has been a keyword for land degradation in the uplands of Vietnam for over a decade. Nevertheless, the "barren" land is still not adequately ecologically characterized. In this work, we analyze land use-induced changes in vegetation and soil properties along a sequence of barren hills types formed on one physiotope. The study is undertaken in the Bac Kan province, one of the poorest upland regions where livestock plays an important role. A transition from an old-growth laurel forest to a sparse manmade grassland is characterized by a total of 177 species, rapid species turnover, and discrete dominants, and an overwhelming effect of disturbance history on both soil and vegetation patterning. Land degradation is most apparent in land use-induced maintenance of arrested successions, and the regeneration course is shifted towards drier formations. We hypothesize a conceptual model as an aid to understanding the process of early fallow differentiation in response to the patterned, fine-scale disturbances. The larger-scale implications of the observed trends in regeneration potentials deviation, and, in particular, the effect of water buffaloes in halting fallow successions, are discussed. PMID- 18392889 TI - Combined MRI and fluoroscopic guided radiofrequency ablation of a renal tumor. AB - Percutaneous CT- and ultrasound-guided radiofrequency ablation of renal cell carcinoma (RCC) has been shown to have very promising medium-term results. We present a unique case of recurrent RCC after partial nephrectomy in a patient with a single kidney and impaired renal function. This tumor could not be visualized either with CT or with ultrasound. A combination of magnetic resonance imaging and fluoroscopic guidance was used, to the best of our knowledge for the first time, to ablate the tumor with radiofrequency. The patient was cancer-free and off dialysis at 30-month follow up. PMID- 18392890 TI - Isolation and characterization of new cyclohexylacetic acid-degrading bacteria. AB - Six cyclohexylacetic acid-degrading strains were isolated from soil samples in Japan and identified as members of the genera Cupriavidus (strain KUA-1), Rhodococcus, and Dietzia by 16S rRNA gene sequence analysis. For the first time members of these genera were shown to be capable of degrading cyclohexylacetic acid. A selected strain, KUA-1, which is the first reported Gram-negative organism capable of growth on cyclohexylacetic acid, was identified as a Cupriavidus metallidurans, based on morphologic and physiologic characteristics and its 16S rRNA gene sequence. Metabolite analysis by HPLC-MS indicated that 1 cyclohexenylacetic acid is an intermediate of cyclohexaneacetic acid metabolism in strain KUA-1. PMID- 18392891 TI - Declination of copper toxicity in pigeon pea and soil system by growth-promoting Proteus vulgaris KNP3 strain. AB - The copper-resistant (1318 microM CuSO(4).5H(2)O) strain KNP3 of Proteus vulgaris was isolated from soil near the Panki power plant, Kanpur, India, and was used to inoculate pigeon pea (Cajanus cajan var. UPS-120) seeds grown in soil for 60 days in the presence of 600 microM CuSO(4).5H(2)O. A study of siderophore production (126.34 +/- 0.52 microg ml(-1)) and its subsequent effects on plant growth promotion under in situ conditions was conducted. The parameters that were monitored included the plants' wet weight, dry weight, shoot length, chlorophyll content, and concentration of copper in plant roots and shoots. The results showed that the strain caused a significant (p < 0.05) increase in wet weight, dry weight, root length, shoot growth, and chlorophyll content (57.8%, 60%, 19.7%, 47.8%, and 36.3%, respectively) in the presence of copper. Furthermore, the strain reduced accumulation of Cu in the roots and shoots to 36.8% and 60.5%, respectively. Apart from this, copper concentration in the soil was measured on 0, 7, 15, 30, and 45 days consecutively and the results indicated that the bioinoculant KNP3 causes a significant decrease in Cu concentration in soil (55.6%), which was unlikely in the control (10.5%) treatment. The data suggested that the bacterial strain has the ability to protect plants against the inhibitory effects of copper besides reducing the copper load of the soil. PMID- 18392892 TI - Easy visualization of the protist Oxyrrhis marina grazing on a live fluorescently labelled heterotrophic nanoflagellate. AB - Planktonic heterotrophic flagellates are ubiquitous eukaryotic microorganisms that play a crucial role in carbon and nutrient fluxes through pelagic food webs. Here we illustrate for the first time a grazing model of planktonic dinoflagellate, Oxyrrhis marina, on the heterotrophic nanoflagellate Goniomonas amphinema, using the DNA-binding fluorescent dye Hoechst 33342. A solution of 1 microg/mL of the fluorochrome allowed viability of the prey for at least 48 hours, provided low fluorescence quenching, and labelled the flagellate without masking the cytoplasm. After 2 hours of contact between the fluorescent prey and the predator, O. marina population had preyed on live G. amphinema at an ingestion rate of 2.2 prey Oxyrrhis (-1) h(-1). Results show that this model is a time-effective and inexpensive approach for the direct observation of heterotrophic flagellate grazing. The fact that prey remain alive while predation occurs, as well as the low rate of quenching, could be of help in studying the fate of real-time trophic interactions between protists in microbial webs. PMID- 18392893 TI - Development of molecular markers of ectomycorrhizal fungi based on ITS region. AB - Four heavy-metal-tolerant isolates of ectomycorrhizal fungi namely Laccaria fraterna (Cooke & Massee) Pegler (EM-1083) and Pisolithus tinctorius (Mich. Ex Pers) Coker & Couch (EM-1290, EM-1293 and EM-1299) were selected on the basis of the previously performed experiments. DNA extraction and purification was done by following standard protocols. Subsequently, polymerase chain reaction (PCR) amplification of the ITS region was done using universal primer ITS 1 and ITS 4. The amplicons were digested with HinfI. The prominent band of the restricted fragments was excised and purified using geneEXIT. Cloning and sequencing of the excised fragment was done following the standard method. Primers were designed with sequence information using the Gene Fisher Interactive PCR Primer design software. Amplification was successfully obtained using the designed primers in Pisolithus tinctorius (EM-1293). The size of the amplicon was 200 bp. PMID- 18392894 TI - Optimising laparoscopic cholangiography time using a simple cannulation technique. AB - BACKGROUND: Opponents of the routine use of intraoperative cholangiography (IOC) express concern over its technical difficulty and the length of time it takes. AIM: To evaluate the impact of our cystic duct cannulation (CDC) technique, as implemented by one consultant and his trainees, on the IOC time. METHODS: IOC is done routinely in all the laparoscopic cholecystectomies (LCs) undertaken in our unit. We carried out a prospective audit over a period of 18 months, recording the IOC time in consecutive patients undergoing laparoscopic cholangiography (LC) with and without laparoscopic common bile duct exploration (LCBDE). The total IOC time was considered to consist of two components: cystic duct cannulation (CDC) time and fluoroscopy time. The IOC time was further analysed according to the difficulty of cannulation and the operator experience. Special consideration was given to the LCBDE cases. We also describe the detailed steps of our CDC technique. RESULTS: Over a period of 18 months 243 patients underwent LC. IOC was completed in 240 patients (98.8% success rate). Of those, 194 were females (81%). The mean age was 50 years (range 18-85 years). The mean total IOC time was 6 min, with a CDC time of 2 min, and fluoroscopy time of 4 min. On further analysis, CDC was considered easy in 86% of cases with a mean CDC time of 1.5 min and total IOC time of 4.3 min. When cannulation was difficult (14% of cases) a cholangiography clamp had to be used to prevent leakage of contrast. In difficult cases, the CDC and IOC mean times were 5 and 8.5 min, respectively. As would be expected, trainees spent more time performing cannulation and completing the IOC than the specialist surgeon (3.8 versus 1.8 min, and 7.2 versus 5.6 min, respectively). These differences were statistically but not clinically significant. Similarly, the IOC time was also significantly increased in LCBDE (13 min). This was mainly due to an increase in fluoroscopy time (10 min) rather than CDC time (3 min). CONCLUSION: The IOC time could be optimised by using a simple and learnable cannulation technique to less than 5 min in most LCs. Surgeons should not, therefore, refrain from using this important investigation on selective or routine basis, subject to their policy for dealing with patients with suspected bile duct stones. PMID- 18392896 TI - Systemic inflammation and cardiovascular risk factors: are morbidly obese subjects different? AB - BACKGROUND: Systemic inflammation is a hallmark of obesity as well as of other chronic diseases, usually indicating increased cardiovascular risk; however, studies with arterial documentation in morbid obesity are extremely scarce. Aiming to analyze correlation between inflammatory markers, pulse-wave velocity (PWV), and intima-media thickness (IMT), a prospective study was designed. METHODS: Morbidly obese patients [n = 29, age 46.3 +/- 5.2 years, 82.8% females (24/29), BMI 44.9 +/- 5.2 kg/m(2)] with C-reactive protein/CRP > 5 mg/l but free from trauma, infection, inflammation, or cancer were enrolled in this study. All were clinically stable candidates for elective bariatric operation. Variables included comorbidities, metabolic profile, inflammatory and coagulatory markers, and arterial morpho-functional indices. RESULTS: Patients suffered from arterial hypertension (72.4%), metabolic syndrome (58.6%), and other comorbidities, but PWV and IMT were less aberrant than expected. Univariate correlation confirmed worse prognosis for those with metabolic syndrome and other accepted clinical risk factors. Multivariate confirmation was achieved for triglycerides (PWV) and D-dimer (IMT), but not for CRP, serum amyloid A, or neutrophil count, which were reversed in certain circumstances. CONCLUSIONS: (1) Metabolic syndrome, hyperglycemia, hypertriglyceridemia and D-dimer were positively correlated with arterial measurements, whereas inflammatory and coagulatory markers often exhibited paradoxical association; (2) stratification confirmed that at certain levels of systemic inflammation or body mass index, acute phase proteins and other markers became unreliable or shifted signals; (3) when controlled for blood pressure, PWV was only moderately elevated, and IMT remained normal; (4) taken together, these findings are consistent with a unique interaction between adiposity, inflammation, and cardiovascular risk in seriously obese subjects. PMID- 18392895 TI - Allelopathic enhancement and differential gene expression in rice under low nitrogen treatment. AB - The allelopathy-competition separation (ACS) based approach was used to explore the biointerference relationship between rice accessions and barnyardgrass exposed to different nitrogen (N) supplies in hydroponics. Rice accession PI312777 exhibited high allelopathic potential to suppress the growth of accompanying weeds, especially when the culture solution had low N content. The non-allelopathic rice Lemont showed an opposite result. Additionally, subtractive hybridization suppression (SSH) was used to construct a forward SSH-cDNA library of PI312777 to investigate gene expression profiles under low N treatment. A total of 35 positive clones from the SSH-cDNA library were sequenced and annotated. According to the function category, 24 genes were classified into five groups related to primary metabolism, phenolic allelochemical synthesis, plant growth/cell cycle regulation, stress response/signal transduction, and protein synthesis/degradation. Among them, two up-regulated genes that encode PAL and cytochrome P450 were selected. Their transcript abundance at low N level was compared further between the allelopathic rice and its counterpart by utilizing real-time quantitative polymerase chain reaction (qRT-PCR). The transcription levels of the two genes increased in both rice accessions when exposed to low N supply, but PI312777 at a higher magnitude than Lemont. At 1, 3, and 7 days of the treatments, the corresponding relative expression levels of PAL were 11.38, 4.83, and 3.57 fold higher in PI312777 root, but there were 1.15, 2.74, and 2.94 fold increases for Lemont, compared with the control plants fed with regular nutrient. The same trend was found for cytochrome P450. These findings suggest that the stronger ability of PI312777 to suppress target weeds, especially in low N nutrient conditions, might be attributed to the stronger activation of the genes that function in de novo synthesis of allelochemicals. PMID- 18392898 TI - Laparoscopic sleeve gastrectomy as an isolated bariatric procedure: intermediate term results from a large series in three Austrian centers. AB - BACKGROUND: Gastric sleeve resection was initially planned as the first step of bilio-pancreatic diversion with duodenal switch but it continues to emerge as a restrictive bariatric procedure on its own. We describe intermediate results in a series of 126 laparoscopic sleeve gastrectomies (LSG) compiled from three bariatric centers in eastern Austria. METHODS: The stomach was laparoscopically reduced to a "sleeve" along the lesser curvature over a 48-Fr bougie. Special attention was placed on complete resection of the gastric fundus. RESULTS: After a mean follow-up of 19.1 months, patients had lost between 2.3 and 27 kg/m(2) or between 6.7% and 130% of their excessive weight. Sixty four percent of the patients lost >50% of their excess weight within an average of 20 months. Seven percent of the patients had an excess weight loss <25% and were therefore considered as failures. The only major surgical complication was leakage of the staple-line needing revision (three times). There were no operative mortalities. CONCLUSION: The final place of LSG in bariatric surgery is still unclear, but our results and those of others show that LSG can be a viable alternative to established procedures. PMID- 18392897 TI - Lipid mobilization following Roux-en-Y gastric bypass examined by magnetic resonance imaging and spectroscopy. AB - BACKGROUND: Recent developments of magnetic resonance imaging (MRI) and spectroscopy have made it possible to quantify lipid deposited in different tissues. To what extent an improvement of glucose tolerance shortly after Roux-en Y gastric bypass surgery (RYGBP) is reflected in lipid levels in liver and skeletal muscle, markers of insulin resistance, has not been clarified. METHODS: Whole-body MRI and MR spectroscopy (MRS) of liver and muscle and measurements of biochemical markers of glucose and lipid metabolism were performed at baseline and 1, 6, and 12 months following surgery in seven morbidly obese women. Volumes of adipose tissue depots and liver and muscle lipids were assessed from the MRI/MRS data. RESULTS: At 1 month postoperatively, body mass index and visceral and subcutaneous adipose tissues were reduced by 9%, 26%, and 10%, respectively, whereas no reductions in intrahepatocellular or skeletal intramyocellular lipid concentrations were found. Free fatty acid and beta-hydroxybutyrate levels were elevated two- and sixfold, respectively; glucose and insulin levels were lowered, indicating increased insulin sensitivity. Further weight loss up to 1 year was associated with reductions in all investigated lipid depots investigated, with the exception of the intramyocellular compartment. CONCLUSION: RYGBP causes rapid lipid mobilization from visceral and subcutaneous adipose depots and enhanced free fatty acid flux to the liver. An exceptional disconnection between liver fat and insulin sensitivity occurs in the early dynamic phase after surgery. However, in the late phase, the energy restriction imposed by the surgical procedure also reduces the liver lipids, but not the intramyocellular lipids. PMID- 18392899 TI - Preoperative weight loss as a predictor of long-term success following Roux-en-Y gastric bypass. AB - BACKGROUND: All patients undergoing gastric bypass surgery at this institution are recommended to achieve a goal of 10% total body weight (TBW) loss prior to surgery. The objective of this study was to determine whether preoperative TBW correlated with 3- and 4-year weight loss outcome. METHODS: This study was conducted prospectively at a large teaching hospital. All adult patients with 3- and 4-year follow-up data since the start of the study in 1998 to September 2007 were included. All data are expressed as mean +/- SD. Pairwise correlation and ordinary least squares regression analysis was used to determine the strength of association between preoperative TBW loss and weight loss at 3 and 4 years. RESULTS: One hundred fifty patients (120 females), age 45.3 +/- 8.9 years, were included. Their body mass indexes (BMIs), preoperatively and after 3 years, were 52.2 +/- 9.8 and 35.4 +/- 8.2 kg/m(2), respectively. There was a significant correlation between preoperative and 3-year TBW lost (9.5 +/- 6.8% vs 31.9 +/- 11.7%, r = 0.302, p = 0.0002) and between excess body weight (EBW) lost preoperatively and after 3 years (16.1 +/- 11.3% vs 55.1 +/- 20.2%, r = 0.225, p = 0.006). Ninety five patients had follow-up data available at 4 years. Their mean preoperative BMI was 52.6 +/- 9.7 kg/m(2) and decreased to 37.5 +/- 9.0 kg/m(2). The TBW loss prior to and after surgery (10.0 +/- 6.5% vs 29.4 +/- 11.5%) was significantly correlated (r = 0.247, p = 0.015). The EBW loss preoperatively and after 4 years correlated positively (17.1 +/- 11.1% vs 50.8 +/ 19.8%, r = 0.205, p = 0.046). CONCLUSION: There is a significant correlation between weight loss attained preoperatively and sustained weight loss at 3 and 4 years. PMID- 18392900 TI - Class III obesity and its relationship with the nutritional status of vitamin A in pre- and postoperative gastric bypass. AB - BACKGROUND: Epidemiological findings have shown the rise of the prevalence of obesity in several segments of the world population, and more recent evidences point to a possible association with vitamin A deficiency (VAD). The aim of this study was to investigate vitamin A nutritional status in individuals with class III obesity in the preoperative period and 30 and 180 days after Roux-en-Y gastric bypass, correlating these findings with lipid profile and body mass index (BMI). METHODS: The sample was composed of class III obese individuals, males and females, in the preoperative period (T1) and 30 (T2) and 180 days (T3) after bariatric surgery with 5,000 IU of supplementation of retinol acetate. Vitamin A nutritional status was assessed through biochemical indicators (retinol and beta carotene serum levels), which were quantified by high-performance liquid chromatography with an inadequacy cutoff of <1.05 micromol/L and >or=40 microg/dL, respectively. RESULTS: Out of the 114 assessed patients, the mean age was 36.9+/-11.6 years and BMI was >or=40 kg/m2. The prevalence of VAD was 14%, being 37.5% in T1, 50.8% and 67.8% in T2, and 52.9% and 67% in T3, according to retinol and serum beta-carotene, respectively. A decrease of retinol and beta carotene serum levels was observed with BMI increase in T1. An increase of very low-density lipoprotein cholesterol (VLDLc) and triglycerides was verified with the increase of retinol, being VLDLc in T3 and triglycerides in T1 and T2. In T1, high-dense lipoprotein cholesterol presented a positive and significant correlation with beta-carotene and a negative and significant correlation in T3. CONCLUSION: High prevalence of VAD in the preoperative period and in 30 and 180 postoperative days, even during supplementation, with higher inadequacy of beta carotene in all the three studied time periods, probably occurred because of its bioconversion to retinol due to the increased demand to which those individuals were exposed. It is suggested that the oral intake supplementation does not present the expected impact, and the need of assessing the nutritional status of vitamin A in the pre- and postoperative Roux-en-Y gastric bypass is emphasized. High prevalence of VAD in T1, with severity in T2 and T3, corroborates the utilization of the cutoff of <1.05 micromol/L as a VAD marker in the studied segment and reinforces its utilization as a tool in clinical practice to identify VAD in morbid obese patients who underwent bariatric surgery. PMID- 18392902 TI - Lap-band causing left gastric artery erosion presenting with torrential hemorrhage. AB - A 61-year-old lady presented 2 years after lap-band surgery with hemetemesis. She was stable on admission and band erosion was diagnosed on gastroscopy. Laparotomy was performed to remove the lap band. Upon division of the lap band, torrential hemorrhage from the eroded left gastric artery was encountered. An anterior gastrostomy was done to expose the artery. Intraoperative gastroscopy was also performed to define the cardioesophageal junction. The artery was ligated and the perforation and gastrostomy were repaired. The patient was discharged after 9 days in hospital. This case highlighted the potential injury to the left gastric artery in patients with band erosion presenting with hemetemesis. Torrential bleeding may be encountered upon division of the lap band, and this should be anticipated during laparotomy. PMID- 18392901 TI - One year after laparoscopic "tight" sleeve gastrectomy: technique and outcome. AB - BACKGROUND: Laparoscopic sleeve gastrectomy (LSG) is currently gaining ground as a new option for the treatment of morbid obesity. The main advantages of this procedure are less postoperative food restrictions, no vomiting, and absence of late complications due to the lack of foreign implants. The aim of this study is to present our experience with this new bariatric technique. METHODS: Ninety three obese patients (65 females and 28 males) who underwent LSG between September 2005 and September 2007 were studied in terms of postoperative complications and weight loss. RESULTS: Mean age was 38.37 +/- 10.81 years (range 19-69) and mean preoperative weight and body mass index (BMI) were 139.12 +/- 24.03 kg (range 100-210) and 46.86 +/- 6.48 kg/m(2) (range 37-66), respectively. Mean follow-up was 12.51 +/- 4.15 months (range 3-24). There were no mortalities, but there were four major and four minor postoperative complications. The mean postoperative excess weight loss (EWL) was 58.32 +/- 16.54%, while mean BMI dropped to 32.98 +/- 6.54 kg/m(2). Mean EWL 3, 6, 12, and 24 months after the operation was 31%, 53%, 67%, and 72%, respectively. Superobese patients (BMI > 50 kg/m(2)) lost less weight. CONCLUSION: In the short term, LSG is a safe and highly effective bariatric operation more suitable for intermediate morbidly obese patients with BMI between 40 and 50 kg/m(2). PMID- 18392903 TI - Gastric migration and strangulation after adjustable gastric banding. AB - We present a case of gastric strangulation 6 months after laparoscopic adjustable gastric banding (LAGB). The 45-year-old woman presented to our emergency department with acute left upper quadrant abdominal pain. Initial upper gastrointestinal studies after emergency department presentation showed no flow through the gastric band and an unusual air/fluid level just above the band, not communicating with the proximal pouch. The patient underwent emergency diagnostic laparoscopy, during which strangulation of a portion of the gastric fundus was identified. During this laparoscopic procedure, the band was removed, and the strangulated portion of stomach was resected using a laparoscopic stapling device. The patient had an uneventful postoperative course and was discharged 6 days after surgery. We present this case as an example of a rare late acute complication resulting from LAGB, which should be recognized and treated surgically on an emergent basis. PMID- 18392904 TI - Reproductive considerations and pregnancy after bariatric surgery: current evidence and recommendations. AB - Obesity has reached epidemic proportions in the USA. Bariatric surgery is an important and increasingly utilized treatment option for morbid obesity refractory to medical therapy. Approximately half of all bariatric surgery patients are reproductive-aged women and, thus, represent a unique patient population with specific concerns. This manuscript focuses on issues of increased postoperative fertility, nutritional monitoring and supplementation, safety of pregnancy after bariatric surgery, and effect of pregnancy on postoperative weight loss. Current recommendations regarding management of patients both before and during pregnancy are provided. In addition, we highlight areas where more research on this issue is needed and advocate for a multidisciplinary approach to patient care. PMID- 18392905 TI - Massive upper gastrointestinal hemorrhage: an unusual presentation after laparoscopic adjustable gastric banding due to erosion into the celiac axis. AB - Laparoscopic adjustable gastric banding (LAGB) is the most common bariatric surgical procedure done currently. It is the preferred choice as it is adjustable, minimally invasive, easily reversible, and does not cause metabolic complications. However, complications like slippage, leakage, erosion-causing perforation, pouch dilatation, pouch herniation, oesophageal dilatation/dysmotility, port disconnection, and migration of band have been reported. We report a rare case of LAGB who presented with life-threatening upper gastrointestinal hemorrhage due to erosion of band into celiac axis 4 months after the operation. An urgent laparotomy was necessary to control the hemorrhage from the celiac axis. PMID- 18392907 TI - Long-term weight regain after gastric bypass: a 5-year prospective study. AB - BACKGROUND: A certain weight gain occurs after obesity surgery compared to the lower weight usually observed between 18 and 24 months postsurgery. The objective of this study was to evaluate weight regain in patients submitted to gastric bypass over a 5-year follow-up period. MATERIALS AND METHODS: A longitudinal prospective study was conducted on 782 obese patients of both genders. Only patients with at least 2 years of surgery were included. The percentage of excess body mass index (BMI) loss at 24, 36, 48, and 60 months postsurgery was compared to the measurements obtained at 18 months after surgery. Surgical therapeutic failure was also evaluated. RESULTS: Percent excess BMI loss was significant up to 18 months postsurgery (p < 0.001), with a mean difference in BMI of 1.06 kg/m2 compared to 12 months postsurgery. Percent BMI loss was no longer significant after 24 months, and weight regain became significant within 48 months after surgery (p < 0.01). Among the patients who presented weight regain, a mean 8% increase was observed within 60 months compared to the lowest weight obtained at 18 months after surgery. The percentage of surgical failure was higher in the superobese group at all times studied, reaching 18.8% at 48 months after surgery. CONCLUSION: Weight regain was observed within 24 months after surgery in approximately 50% of patients. Both weight regain and surgical failure were higher in the superobese group. Studies in regard to metabolic and hormonal mechanisms underlying weight regain might elucidate the causes of this finding. PMID- 18392906 TI - Management of anastomotic leaks after laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: Anastomotic leaks after bariatric surgery carry high morbidity and mortality. We aimed to describe our experience of the diagnosis and management of gastrointestinal anastomotic leaks in patients undergoing laparoscopic gastric bypass in a single institution. METHODS: Of 1,200 patients who underwent laparoscopic Roux-en-Y gastric bypass with manual gastrojejunal anastomosis for morbid obesity from January 2002 to January 2007, we retrospectively analyzed 59 patients with anastomotic leak. The location of the leak, day of diagnosis, diagnostic methods, clinical manifestations, treatment modalities, associated complications, and length of hospital stay were analyzed. RESULTS: Leaks were located as follows: 67.8% in the gastrojejunostomy, 10.2% in the gastric pouch, 3.4% in the excluded stomach, 5.1% in the jejunojejunal anastomosis, 3.4% in the gastrojejunostomy plus pouch, 3.4% in the pouch plus excluded stomach, and 6.8% in undetermined sites. Routine upper gastrointestinal series revealed contrast extravasation in nine patients (15.3%). Leaks were asymptomatic at diagnosis in 29 patients (49.2%). Surgical reintervention was carried out in 23 patients, and conservative treatment was provided in the remaining 36. Transfer to the intensive care unit was required in 11 patients, with five deaths (0.4%). CONCLUSION: In our experience, most anastomotic leaks can be managed with conservative measures alone. In many patients, abdominal drains are effective in the management of leaks, obviating the need for reintervention. Nasoenteral nutrition was effective in the non-operative management of gastrojejunal leaks in patients without signs of systemic toxicity. PMID- 18392908 TI - Combined laparoscopic gastric banding and stomach reduction (GBSR): initial experience after 1 year. AB - PURPOSE: We combined laparoscopic adjustable gastric banding (LAGB) and laparoscopic partial gastrectomy into a single procedure (GBSR). METHODS: The 6- and 12-month percent excess weight loss (EWL) was compared to results expected from the literature for LAGB and laparoscopic Roux-en-Y gastric bypass (LRYGB) using t test. The time to band adjustment was compared with the 6-week period experienced in our setting for LAGB using a sign test. RESULTS: Eighteen patients were considered. One patient had gastric leak and had the band removed in postoperative day 1. Seventeen patients were followed-up. The patients, 4 men and 13 women, had a mean age of 38.8 years, an initial mean BMI of 52.4 kg/m2, and a mean of 8.3 comorbidities. All required outpatient band adjustment, eight at 4 months, eight at 5 months, and one at 6 months; the median 5 months was greater than the 6 weeks expected for LAGB (P < 0.001). The mean 6-month EWL, 28.6%, was similar to that expected for LAGB (P = 0.24) and less than that expected for LYRGB (P < 0.001). The mean 12-month EWL, 57.4%, was greater than that expected for LAGB (P < 0.001) and less than that expected for LYRGB (P < 0.001). CONCLUSION: Although GBSR's EWL at 6 and 12 months is less than that of LRYGB, GBSR should be further studied and applied in specific situations only. PMID- 18392911 TI - Crucial role of hand surgery in orthopedic surgery in Japan. PMID- 18392910 TI - The assessment of quality of life in a trial on lightweight mesh fixation with fibrin sealant in transabdominal preperitoneal hernia repair. AB - BACKGROUND: Chronic pain is a major concern in open and laparoscopic hernia repair. Study groups have adopted a variety of tools to assess postoperative (postOP) pain and quality of life (QoL). Unfortunately, modifications of existing tests and self-designed questionnaires are common, yielding unvalidated results and making comparison of data difficult. The aim of this study was to assess the QoL in transabdominal preperitoneal mesh repair (TAPP) with fibrin sealant (FS) for lightweight mesh fixation, applying the standardised Short Form 36 (SF36) questionnaire in its unmodified design. The SF36 has already been validated and implemented in a large number of studies. In this trial the physical-health component summary measure (PHM), summarising the physical health-related scales, served as the primary outcome parameter. MATERIALS AND METHODS: After informed written consent was obtained, TAPP with fibrin mesh sealing was performed in 11 non-selected consecutive patients by a single surgeon. A direct control group (e.g. TAPP with staples) was not enrolled, because a favourable change in the QoL in patients subjected to the mesh sealing approach was the tested hypothesis and not the comparison of techniques. The macroporous mesh (TI-Mesh, GfE, Germany) was fixed with 1 ml of FS (FS, Tisseel, Tissucol, Baxter Biosciences, Austria), and the QoL and pain were assessed preoperatively and 1 year postOP using the SF36 survey and the visual analogue score (VAS). RESULTS: After 12 months, recurrences or complications were observed. The analysis of the unmodifed SF36 revealed a highly significant improvement in the PHM, based on significant changes of all physical-health-related scales. The scale 'social functioning' (SOCIAL), which belongs to the mental-health-related scale, had also significantly improved. The VAS was significantly reduced after 1 year. CONCLUSIONS: Despite a small number of patients (n = 11), a strikingly significant improvement in physical health and reduction of pain was detected with the unmodified SF36 and the VAS 1 year after TAPP repair with fibrin-sealed lightweight meshes. We suggest the use of the unmodified SF36 for QoL in hernia repair in order to assess all aspects of recovery (physical and mental) and to facilitate comparison of data. PMID- 18392909 TI - Do modern techniques improve core decompression outcomes for hip osteonecrosis? AB - Core decompression procedures have been used in osteonecrosis of the femoral head to attempt to delay the joint destruction that may necessitate hip arthroplasty. The efficacy of core decompressions has been variable with many variations of technique described. To determine whether the efficacy of this procedure has improved during the last 15 years using modern techniques, we compared recently reported radiographic and clinical success rates to results of surgeries performed before 1992. Additionally, we evaluated the outcomes of our cohort of 52 patients (79 hips) who were treated with multiple small-diameter drillings. There was a decrease in the proportion of patients undergoing additional surgeries and an increase in radiographic success when comparing pre-1992 results to patients treated in the last 15 years. However, there were fewer Stage III hips in the more recent reports, suggesting that patient selection was an important reason for this improvement. The results of the small-diameter drilling cohort were similar to other recent reports. Patients who had small lesions and were Ficat Stage I had the best results with 79% showing no radiographic progression. Our study confirms core decompression is a safe and effective procedure for treating early stage femoral head osteonecrosis. PMID- 18392912 TI - Interleukin-1beta gene polymorphism associated with radiographic signs of osteoarthritis of the knee. AB - BACKGROUND: Osteoarthritis is recognized as a noninflammatory, progressive condition, the principal cause of which is regressive changes associated with aging and which pursues a chronic course. Recently, the involvement of genetic factors has been widely reported. The purpose of this study was to identify polymorphisms at particular risk of osteoarthritis of the knee for community living middle-aged and elderly people. METHODS: Focusing on 359 participants (ages 44-86 years) of the comprehensive health examination program (CHEP), we investigated the presence/absence of radiographic osteoarthritis (ROA) of the knee, and 11 types of gene polymorphisms and their association with ROA. RESULTS: Interleukin-1beta (IL1B) T-31C polymorphism was found to be associated with ROA. In the case of IL1B T-31C polymorphism in the ROA group, a significant difference was found between the groups combining the C/C genotype, the C/T genotype, and the T/T genotype. In particular, the genotypes with the C allele differed from the T/T genotype, with the morbidity rate being higher in the T/T group (odds ratio (OR) 2.04, 95% confidence interval (95%CI) 1.05-3.98, P = 0.036). CONCLUSION: Our results confirm that in IL1B T-31C with the T/T genotype, the rate of ROA was significantly higher than that with the C/C and C/T genotypes. It might be possible to implement active preventative measures, such as avoidance of obesity and excessive exercise loads, for carriers of IL1B T/T. PMID- 18392913 TI - Effects of dry needling at tender points for neck pain (Japanese: katakori): near infrared spectroscopy for monitoring muscular oxygenation of the trapezius. AB - BACKGROUND: Neck pain (katakori) is a common symptom in adult Japanese people. However, the pathophysiological aspect of this condition has not been well documented to date. The purpose of this study was to investigate the effects of tender point dry needling to the trapezius muscles and the resultant changes in muscular hemodynamics. METHODS: "Neck pain" patients were defined as those complaining of dull pain or discomfort mainly along the trapezius muscles without serious spinal or shoulder disorders. We used near-infrared spectroscopy to monitor the changes of oxyhemoglobin (oxyHb) and deoxyhemoglobin (deoxyHb) of the trapezius muscles and a Visual Analogue Scale (VAS) to assess subjective neck pain intensity. Experiment I: Nine subjects with "neck pain" and four control subjects were recruited. Total hemoglobin (Hb) and SdO2 [= oxyHb/(oxyHb + deoxyHb)] were measured before and immediately after needling for 15 min. We compared these parameters and VAS before and immediately after needling. Experiment II: Thirteen subjects with "neck pain" were instructed to perform isometric contraction of their trapezius muscles for 1 min; the half-recovery time of SdO2 (defined as T(R)) was measured. After that, all subjects underwent needling. On the next day, we repeated the measurements of T(R) after the same voluntary contraction of the trapezius muscle in the same patients. We compared T(R) and VAS before and on the day after needling. RESULTS: Experiment I: All subjects with "neck pain" reported significant pain relief (P = 0.0147) measured by VAS immediately after needling, but total Hb and SdO2 exhibited no significant change after needling. Experiment II: T(R) was shortened on the day after needling in 10 of 13 patients (P = 0.0043), and neck pain was decreased in 12 patients (P = 0.0158). CONCLUSIONS: After dry needling, total Hb and SdO2 did not change in real time, but T(R) was shortened on the next day. These results showed that the shortening of T(R) would provide a measure by which to assess the effectiveness of treatment for neck pain. PMID- 18392914 TI - Anterior plating and percutaneous iliosacral screwing in an unstable pelvic ring injury. AB - BACKGROUND: This study was carried out to evaluate the effectiveness of anterior plating with subsequent percutaneous iliosacral screwing for the management of unstable pelvic ring injuries. METHODS: Nineteen patients with unstable pelvic ring injuries were included in this retrospective study. All patients were followed up for at least 1 year, and their mean age was 43 years. According to the Association for Osteosynthesis-Orthopaedic Trauma Association (AO-OTA) classification, there were 5 B2 injuries, 11 C1 injuries, and 3 C2 injuries. After anterior fixation by means of plating, an iliosacral screw fixation was carried out percutaneously using a C-arm fluoroscope. RESULTS: All fractures healed, except for 1 case of nonunion at the pubic ramus. Radiological results showed that there were 9 anatomic, 7 nearly anatomic, 2 moderate, and 1 poor reduction. Sixteen of the 19 patients had good or excellent results for function, and all these had satisfactory (anatomic or nearly anatomic) reductions. The two moderate and 1 poor result were from an unsatisfactory reduction in a type-C injury with residual neurological signs. A screw misplacement with a neurological compromise occurred in 1 patient, but there were no adverse sequelae after its removal. The complications encountered were 2 cases of screw loosening, 2 cases of anterior metal failure, and 1 deep infection. CONCLUSIONS: Anterior plating with subsequent percutaneous iliosacral screwing may be a useful method of treatment for unstable pelvic ring injuries, and the reduction quality and residual neurological signs were important in its functional outcome. PMID- 18392915 TI - Quality of life following femoral osteotomy and total hip arthroplasty for nontraumatic osteonecrosis of the femoral head. AB - BACKGROUND: Nontraumatic osteonecrosis of the femoral head (NOFH) frequently develops in active young persons. The affected femoral head collapses owing to weight-bearing, and the individual's quality of life (QOL) can be predicted to deteriorate greatly with time. We undertook to determine the efficacy of surgery and to clarify whether patient QOL differs according to differences in the surgical method employed. METHODS: We cross-sectionally compared QOL in NOFH patients treated with femoral osteotomy, total hip arthroplasty (THA), or nonoperatively. A total of 81 cases were available for study, comprising 41 with osteotomy, 19 with THA, and 21 in the nonoperative group. The mean age was significantly higher in the THA group than in the other two groups. The Japanese Orthopaedic Association (JOA) hip score and Visual Analogue Scale (VAS) regarding hip pain were compared among the groups. These groups were also analyzed for their health-related QOL using the Short Form Health Survey (SF-36) with analysis of variance for age adjustment. RESULTS: The mean JOA score was significantly lower in the nonoperative group than in the osteotomy group. The mean VAS scores showed no significant difference between any of the three groups. Regarding the subscales of SF-36, the physical functioning subscale in the nonoperative group showed a significantly lower value than was seen in the osteotomy group (P = 0.003). The physical component summary (PCS) scores were 39.4 (osteotomy group), 39.1 (THA group), and 27.8 (nonoperative group), with a significant difference between the osteotomy and nonoperative groups (P = 0.027). There was also a trend for a better PCS scores in the THA group than in the nonoperative group (P = 0.056). The mental component summary scores were 49.6 (osteotomy group), 50.3 (THA group), and 48.3 (nonoperative group), with no significant difference found among any of the three groups. CONCLUSIONS: Among patients with NOFH, physical function impairment was a more potent factor than pain for decreasing QOL in the nonoperative group than in the surgical groups. Furthermore, osteotomy and THA were similar in regard to the evaluation of the postoperative QOL score if the indications for osteotomy were strictly applied. PMID- 18392916 TI - Anatomical analysis of the anterior cruciate ligament femoral and tibial footprints. AB - BACKGROUND: The current trend in anterior cruciate ligament (ACL) reconstruction has shifted to anatomical double-bundle (DB) reconstruction, which reproduces both the anteromedial bundle (AMB) and the posterolateral bundle (PLB) of the ACL. Navigation systems have also been recently introduced to orthopedic surgical procedures, including ACL reconstruction. In DB-ACL reconstruction, the femoral and tibial tunnel positions are very important, but a representation of the ACL footprint under an arthroscopic view has not been established even though navigation systems have been introduced. The purpose of this study was to evaluate the anatomical footprints of both the AMB and the PLB using the representation method for application to arthroscopic DB-ACL reconstruction using a navigation system, and to evaluate the validity of the currently determined footprint position compared with other representation methods. METHODS: Thirty six cadaveric knees were used for an anatomical evaluation of footprints of the AMB and PLB. On the tibial side, the ACL footprints were evaluated using an original method. On the femoral side, the ACL footprints were evaluated using Watanabe's method and three other methods: (1) the quadrant method, (2) Mochizuki's method, and (3) Takahashi's method. RESULTS: The central points of the ACL footprints were represented almost constantly. The present data is in accordance with previous measurement data. CONCLUSION: This study showed that the anatomical data of the ACL femoral and tibial footprints determined with Watanabe's method at the femoral side and our original method at the tibial side were both applicable to arthroscopic surgery with a navigation system. PMID- 18392917 TI - Fas ligand expression on human nucleus pulposus cells decreases with disc degeneration processes. AB - BACKGROUND: The intervertebral disc has been reported to be an immunologically privileged environment, possibly mediated by Fas ligand (FasL) expression. On the other hand, recent studies have shown the infiltration of host immune cells into the degenerated disc, which may indicate the failure of the immune-privilege feature of the disc with degeneration. However, the relationship between FasL expression and disc degeneration is still unclear. Therefore, the purpose of this study was to clarify the relationship between FasL expression and disc degeneration. METHODS: Ten human degenerated disc specimens were obtained from spondylolisthesis patients and ten nondegenerated discs from idiopathic scoliosis patients during surgical procedures. Immunohistochemical staining was performed to determine the presence of FasL in cross-sections of those discs. Parts of the disc tissues were used to examine FasL expression quantitatively with Western blot analysis. To examine whether the change in FasL expression was influenced by aging, an animal study comparing the discs from young and old rats were performed using magnetic resonance imaging (MRI) and real-time polymerase chain reaction (PCR) assessment. RESULTS: Nucleus pulposus cells showed strong positive staining for FasL in all specimens examined. Quantitative examination demonstrated a significant decrease in FasL expression in the degenerated group compared with the nondegenerated group (average 67.6%, P<0.05). MRI showed no significant differences in the grade of disc degeneration between young and old rats, and also no significant difference in FasL mRNA in real-time PCR assay. CONCLUSIONS: The current results indicate that FasL and its potential mechanism of immunological privilege could influence the protection of the intervertebral disc against degeneration. PMID- 18392918 TI - Effects of calcitonin on the biomechanics, histopathology, and radiography of callus formation in rats. AB - BACKGROUND: This study was designed to examine the effect of salmon calcitonin on fracture repair. METHODS: A transverse middiaphyseal fracture of the right tibia was surgically induced, and stabilized by an intramedullary Kirschner wire. Eighty male Wistar rats were arbitrarily assigned to four groups of 20 animals each. Groups I and II were the controls and did not receive any medication but did receive placebo injections. The other two groups received 5 IU/kg/day salmon calcitonin intramuscularly for 6 weeks (Group III) or for 10 weeks (Group IV) postoperatively. The formation and healing of the bones were determined by radiographic and histopathological analyses and by biomechanical tests. RESULTS: In radiographic examinations, there were no statistically significant differences between groups I and III at week 6 or between groups II and IV at week 10. However, the histopathological evaluation scores were higher in the calcitonin group at the early stage (6 weeks) of fracture healing, which indicates a more mature callus formation (P < 0.05). The values for maximum torsional moment during fracture were higher in the calcitonin group in both stages of fracture healing (P < 0.01). CONCLUSIONS: This study could have two important clinical implications. (1) Higher scores for the histopathological evaluation and a greater resistance to moment force applied at an early stage (week 6) of fracture healing imply that calcitonin intake might enable us to allow earlier mobilization and weight-bearing in clinical cases with rigid fixation. (2) At a late stage of fracture healing (week 10), the significantly better (P < 0.0001) results obtained in the biomechanical parameters used might imply that calcitonin intake could enable us to perform early implant removal, and strongly suggest that the strength and quality of the callus formation could be improved by administering calcitonin following a fracture. PMID- 18392919 TI - Revision high tibial osteotomy with the Taylor spatial frame for failed opening wedge high tibial osteotomy. PMID- 18392920 TI - Cerebellar hemorrhage after spine surgery. PMID- 18392921 TI - Fatal prognosis of an atypical meningioma in the cervical spine. PMID- 18392922 TI - Microbiology and management of joint and bone infections due to anaerobic bacteria. AB - PURPOSE: To describes the microbiology, diagnosis, and management of septic arthritis and osteomyelitis due to anaerobic bacteria. RESULTS: The predominant anaerobes in arthritis are anaerobic Gram-negative bacilli (AGNB) including the Bacteroides fragilis group, Fusobacterium spp., Peptostreptococcus spp., and Propionibacterium acnes. Infection with P. acnes is associated with a prosthetic joint, previous surgery, and trauma. B. fragilis group is associated with distant infection, Clostridium spp. with trauma, and Fusobacterium spp. with oropharyngeal infection. Most cases of anaerobic arthritis, in contrast to anaerobic osteomyelitis, involved a single isolate, and most cases are secondary to hematogenous spread. The predominant anaerobes in osteomyelitis are Bacteroides, Peptostreptococcus, Fusobacterium, and Clostridium spp. as well as P. acnes. Conditions predisposing to bone infections are vascular disease, bites, contiguous infection, peripheral neuropathy, hematogenous spread, and trauma. Pigmented Prevotella and Porphyromonas spp. are mostly isolated in skull and bite infections, members of the B. fragilis group in hand and feet infections, and Fusobacterium spp. in skull, bite, and hematogenous long bone infections. Many patients with osteomyelitis due to anaerobic bacteria have evidence of an anaerobic infection elsewhere in the body that is the source of the organisms involved in the osteomyelitis. Treatment of arthritis and osteomyelitis involving anaerobic bacteria includes symptomatic therapy, immobilization in some cases, adequate drainage of purulent material, and antibiotic therapy effective against these organisms. CONCLUSIONS: Anaerobic bacteria can cause septic arthritis and osteomyelitis. Correct diagnosis and appropriate therapy are important contributor to successful outcome. PMID- 18392923 TI - Use of palliative radiotherapy trials for clinical biomarker development. AB - INTRODUCTION: Approximately one quarter of all cancer patients will require palliative radiation treatment at some point during the course of their disease, but only a minority of these patients are entered in clinical trials. ETHICAL ASSESSMENT OF BIOMARKERS IN PALLIATIVE RADIOTHERAPY TRIALS: We review the literature debating the ethics of inclusion of "palliative" patients on clinical trials. We suggest that these patients provide a potentially valuable resource that can be leveraged to facilitate the discovery and validation of biomarkers predictive of radiation response and toxicity. In addition, this patient population offers valuable opportunities to test combination of radiation and targeted therapies to screen for activity, toxicity and biomarkers in a relatively safe manner. CONCLUSION: Patients undergoing palliative radiation therapy may provide new opportunities for the development and testing of predictive radiotherapy biomarkers as well as affording opportunities to test combinations of radiation and targeted therapies. PMID- 18392924 TI - General and specific self-efficacy reports of patients with chronic low back pain: are they related to performances in a functional capacity evaluation? AB - INTRODUCTION: The objective of this study was to analyze the relationship of general and specific self-efficacy (SE) beliefs with functional capacity evaluation (FCE) performances in patients with chronic non-specific low back pain (CLBP), while controlling for influence of gender, age, and self-reported pain intensity, self-esteem, disability, psychosocial distress and health status. METHODS: Included were 92 patients with CLBP referred to an outpatient university based multidisciplinary pain rehabilitation program in The Netherlands. All patients underwent an FCE. General SE was measured with the ALCOS questionnaire prior to the FCE, specific SE was measured with a self-constructed standardized question during the FCE. Paired samples t-tests were used to tests differences between predicted and actual performances. Pearson and Spearman rank correlation coefficients were used to express the strength of the relationships between SE and performances. Multivariate analyses were used to test the influence of control variables on the relationships between SE (general or specific) and performances. RESULTS: Performances were consistently higher than patients' self predictions. Differences between predictions and performances were significant in male lifting low, male carrying, and female carrying. With exception of the association between specific SE and lifting in males (r = 0.55, P < 0.05), all other correlations between general and specific SE and FCE performances were non significant. Multivariable regression analyses showed that the relative contribution of SE measures over gender was little or none. CONCLUSIONS: The contribution of specific SE to the prediction of FCE performances is moderate in one instance, and insignificant in most instances (both specific and general SE). Because of the consistency of the differences between prediction (specific SE) and performances, and depending on the level of accuracy needed, future research may deliberate the use of predicted material handling capacities at group level and correct for a systematic underprediction. PMID- 18392925 TI - How well do return-to-work interventions for musculoskeletal conditions address the multicausality of work disability? AB - INTRODUCTION: The best-documented return-to-work rehabilitation programs concern workers with musculoskeletal disorders (MSKD). For this clientele, a global perspective has been adopted which explains the multicausality of work disability. This perspective of work disability proposes that return-to-work interventions should address three central elements: individual psychological factors, work environmental factors and factors related to the involvement of the various stakeholders. Long-term work disability is no longer seen simply as the consequence of impairment, but rather as the result of interactions between the worker and main systems: the health care, work environment and financial compensation systems. METHODS: This paper presents a descriptive content analysis of return-to-work interventions delivered to workers with MSKD which consider this global perspective and which are found to be effective in systematic reviews of the literature. RESULTS: The review of programs designed for workers with MSKD showed that eleven programs address the individual clinical and psychological factors, work environmental factors and factors related to the involvement of the various stakeholders, but in different ways. Only two programs met the essential components identified by the literature. These essential components are: centralized coordination of the worker's return to work, formal individual psychological and occupational interventions, workplace-based interventions, work accommodations, contact between the various stakeholders and interventions to foster concerted action. CONCLUSIONS: Interventions which involve the work environment and concerted action by the various partners seem to require the most investment in terms of energy. The establishment of common principles and shared values regarding work rehabilitation as well as less divided mechanisms for action among the various partners should be considered. PMID- 18392926 TI - Discriminating between individuals with and without musculoskeletal disorders of the upper extremity by means of items related to computer keyboard use. AB - INTRODUCTION: Identifying postures and behaviors during keyboard use that can discriminate between individuals with and without musculoskeletal disorders of the upper extremity (MSD-UE) is important for developing intervention strategies. This study explores the ability of models built from items of the Keyboard Personal Computer Style instrument (K-PeCS) to discriminate between subjects who have MSD-UE and those who do not. METHODS: Forty-two subjects, 21 with diagnosed MSD-UE (cases) and 21 without MSD-UE (controls), were videotaped while using their keyboards at their onsite computer workstations. These video clips were rated using the K-PeCS. The K-PeCS items were used to generate models to discriminate between cases and controls using Classification and Regression Tree (CART) methods. RESULTS: Two CART models were generated; one that could accurately discriminate between cases and controls when the cases had any diagnosis of MSD-UE (69% accuracy) and one that could accurately discriminate between cases and controls when the cases had neck-related MSD-UE (93% accuracy). Both models had the same single item, "neck flexion angle greater than 20 degrees ". In both models, subjects who did not have a neck flexion angle of greater than 20 degrees were accurately identified as controls. CONCLUSIONS: The K-PeCS item "neck flexion greater than 20 degrees " can discriminate between subjects with and without MSD-UE. Further research with a larger sample is needed to develop models that have greater accuracy. PMID- 18392927 TI - Changing community readiness to prevent the abuse of inhalants and other harmful legal products in Alaska. AB - This paper presents results from an application of the Community Readiness Model (CRM) as part of a multi-stage community mobilization strategy to engage community leaders, retailers, parents, and school personnel in preventing youth use of inhalants and other harmful legal products in rural Alaska. The CRM is designed to assess readiness to address a single social problem, based on a limited set of key informant interviews. In this study, researchers conducted 32 baseline and 34 post-intervention community readiness assessment interviews in four rural Alaskan communities. These interviews with key informants from the communities were coded and analyzed using CRM methods to yield readiness scores for each community. The aggregate results were analyzed using hierarchical linear modeling (HLM), and the individual community scores were analyzed in the context of the overall study. Significant positive changes in community readiness were found across six readiness dimensions as well as for the overall readiness score. Variation in the degree of changes in readiness across the four communities is attributed to differences in the intervention's implementation. The implications of these results include the potential for CRM assessments to serve as an integral component of a community mobilization strategy and also to offer meaningful feedback to communities participating in prevention research. PMID- 18392928 TI - Troponin-I elevation in a young man with arrhythmogenic right ventricular dysplasia/cardiomyopathy. AB - Ventricular arrhythmias occur frequently in patients with arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) as well as those with ischemic heart disease. We present the case of a 29-year-old man with ARVD/C and multiple episodes of symptomatic ventricular tachycardia terminated by implantable cardioverter defibrillator (ICD) discharges. Phasic elevation of troponin-I prompted repeated coronary angiograms, all of which were normal. The patient was successfully treated with radiofrequency ablation. This case illustrates that ARVD/C may result in elevated cardiac enzymes in the absence of coronary artery disease. PMID- 18392929 TI - Pacemaker and ICD leads: strategies for long-term management. AB - Pacemaker and defibrillator therapy is on the rise as a result of expanding indications. Unfortunately, this trend is associated with an increased number of cardiac device-related complications. Lead failure, device infection and vascular complication are not uncommon and may cause significant patient morbidity and mortality. Furthermore, the considerable variability in the approach to dealing with device-related complications not infrequently leads to additional problems and complications. This review is intended to provide general guidelines and strategies for long-term management of cardiac devices. Proper implantation techniques that focus on primary prevention of complications, as well as, planning for future procedures are essential. The use of sterile techniques and implementation of peri-operative antibiotics are well supported in clinical trials. Additionally, minimizing the amount of implanted hardware and limiting the procedure length have also been shown to decrease the rates of infectious complications. Once device infection is confirmed, the primary objective, short of a few exceptions, should consist of the entire system removal via open surgery or percutaneously and antibiotic therapy before reimplantation. Vascular occlusions are not uncommon; however, in the majority of cases they are asymptomatic. Nonetheless, they pose a significant problem with device reimplantation, especially in the younger patients who will require multiple generator exchanges in the future. Site preservation should, therefore, become the primary objective. This can also be accomplished with the open surgical or percutaneous device removal. The latter is very safe and effective when performed in experienced centers. Finally, lead follow-up must be routinely conducted to identify problematic leads, prevent further reimplantations, and guide future research. PMID- 18392930 TI - Betablockers: is the reduction of sudden death related to pure electrophysiologic effects? PMID- 18392932 TI - Microsatellite isolation and characterization in Japanese persimmon (Diospyros kaki). PMID- 18392931 TI - Theoretical analysis of reverse-time correlation for idealized orientation tuning dynamics. AB - A theoretical analysis is presented of a reverse-time correlation method used in experimentally investigating orientation tuning dynamics of neurons in the primary visual cortex. An exact mathematical characterization of the method is developed, and its connection with the Volterra-Wiener nonlinear systems theory is described. Various mathematical consequences and possible physiological implications of this analysis are illustrated using exactly solvable idealized models of orientation tuning. PMID- 18392933 TI - Preliminary evidence for the integration of music into HIV prevention for severely mentally ill Latinas. AB - BACKGROUND: Individuals with severe mental illness (SMI) may be at increased risk of HIV infection. Prevention programs designed specifically for SMI have relied primarily on principles of cognitive-behavior change theories delivered in a small group format and in venues and services utilized by SMI. Most intervention effects have not been shown to be sustainable over time. We report on our findings relating to the importance of music to Puerto Rican women with SMI and the implications for HIV prevention interventions with this population. METHODS: We interviewed and shadowed over a 2-year period 53 women of Puerto Rican ethnicity between the ages of 18 and 50, residing in northeastern Ohio, who had been diagnosed with schizophrenia, bipolar disorder, or major depression. RESULTS: Nearly one-half of the participants listened to music regularly. Some reported that music was essential to their lives. Participants reported that music improved their mental and social well-being by facilitating expression and reflection of their emotions and increasing their energy levels. DISCUSSION: Music may affect the core negative symptoms and compensate for neuropsychological deficits in women with schizophrenia and related conditions by facilitating the articulation of emotion and allowing individuals to better attend to and potentially incorporate external activities into their lives. The use of music in HIV prevention efforts with SMI Latinas may facilitate their emotional expression and assist them in integrating the educative efforts into their life style choices. PMID- 18392934 TI - Mammography screening among Arab American women in metropolitan Detroit. AB - Mammography screening behavior has not been well studied among Middle Eastern immigrant women. We conducted a telephone survey of 365 Arab American women residing in metropolitan Detroit, home to one of the largest populations of Middle Eastern immigrants in the US, to determine prevalence of factors associated with mammography, and attitudes and beliefs regarding mammography screening. Of 365 participants, only five were born in the US. Mean age was 53.2 years (SD 10.8). Two hundred twelve (58.1%) reported having mammogram every 1-2 years; 70% ever had mammogram. Age 50-64 years, having health insurance, married status, being in the US over 10 years, and being Lebanese were associated with mammography every 1-2 years. After adjusting for demographic factors, perceived seriousness of disease, general health motivation, and having fewer barriers were associated with more frequent screening. Appropriate mammography screening is decreased in this group. Targeted outreach regarding screening is appropriate for this population; however, lack of insurance may prevent adequate follow-up. PMID- 18392935 TI - Disseminating health information and diabetes care for Latinos via electronic information kiosks. AB - This study investigated the use of electronic, web-enabled touch-screen information kiosks as a tool to provide culturally and linguistically appropriate diabetes information to Latino audiences. Two kiosk models (high privacy sit down, group enabled stand-up) in two locations (pharmacy, community center) in Northeast Georgia provided bilingual, read-aloud diabetes education and local resource information on health care. Data from public use and interviews with focus group participants showed that users found the kiosks and their functions helpful and usable, though usage was moderated by presence or absence of a peer health educator (promotora); participants also preferred the sit-down, multi function kiosk model. PMID- 18392936 TI - Experiential health from an ageing and migration perspective: the case of older Finland-Swedes. AB - Research has shown that immigrants and minority groups tend to have a lower health status compared to the majority population. The Finnish immigrants in Sweden are no exception. The Finland-Swedes, i.e., persons living in Finland who have Swedish as their mother language, seem to be an exception, however. They have been found to have better health and longer life expectancy compared to the Finnish majority. Research on health among migrated Finland-Swedes is scarce. The aim of this study was to describe and deepen the understanding of how older Finland-Swedes living as immigrants in Sweden, as well as re-migrants in Finland, experienced their health. Data was collected through 39 qualitative interviews with 29 older Finland-Swedes aged 65 or more. Data was analysed through qualitative thematic content analysis. The analysis resulted in five themes: Ageing means becoming frail and closer to death; Despite frailty and old age it is possible to feel well and experience peace; Being grateful for health as a source of life; Health comes from inner strength and external sources; Migration meant a mental and physical burden to health. Overall, both ageing and migration were experienced as jeopardising health. PMID- 18392937 TI - Fetal bovine serum simultaneously stimulates apoptosis and DNA synthesis in premeiotic stages of spermatogenesis in spiny dogfish (Squalus acanthias) in vitro: modulation by androgen and spermatogenic activity status. AB - Using the simple cystic spermatogenesis in the shark testis as a model, we previously reported the relative resistance of immature spermatogonia (stem cell and early-stage spermatogonia) to apoptosis in the normal testis and after spermatoxicant exposure in vivo. Apoptosis was monitored by fluorescence image analysis of living cysts, using the validated acridine orange (AO) vital staining technique. Findings show that FBS simultaneously stimulates both apoptosis and [(3)H]thymidine incorporation in immature spermatogonial clones in a concentration-dependent manner in vitro. Furthermore, androgen inhibits apoptosis and increases cyst viability, more so with 10% FBS than with 1% FBS. All the effects were as a function of spermatogenic activity status but were distinct in early-stage spermatogonial cysts isolated from testes awakening from the previous winter spermatogenic arrest period. Results are discussed in the context of the alternating germ-Sertoli cell population kinetics of early-stage spermatogonial cysts in Squalus acanthias's protracted testicular cycle. PMID- 18392938 TI - Protein kinase A-mediated CREB phosphorylation is an oxidant-induced survival pathway in alveolar type II cells. AB - Oxidant stress plays a role in the pathogenesis of pulmonary diseases, including fibrotic lung disease and cancer. We previously found that hydrogen peroxide (H2O2) initiates an increase in Ca2+/cAMP-response element binding protein (CREB) phosphorylation in C10 alveolar type II cells that requires activation of extracellular regulated kinases 1/2 (ERK1/2). Here, we investigated the role of crosstalk between protein kinase A (PKA) and epidermal growth factor receptor (EGFR) in oxidant-induced signaling to ERK1/2 and CREB in C10 cells. Application of H2O2 increased nuclear accumulation of PKA, and inhibition of PKA with H89 reduced oxidant-mediated phosphorylation of both CREB and ERK1/2. Single cell measurements of cAMP and redox status, using a FRET-based biosensor and a redox sensitive GFP, respectively, indicated that H2O2 increases production of cAMP that correlates with redox state. Inhibition of EGFR activity decreased both H2O2 induced CREB phosphorylation and translocation of PKA to the nucleus, suggesting that crosstalk between PKA and EGFR underlies the oxidant-induced CREB response. Furthermore, knockdown of CREB expression using siRNA led to a decrease in bcl-2 and an increase in oxidant-induced apoptosis. Together these data reveal a novel role for crosstalk between PKA, ERK1/2 and CREB that mediates cell survival during oxidant stress. PMID- 18392941 TI - The parapatric distribution and contact zone of two forms showing different male to-male aggressiveness in a social spider mite, Stigmaeopsis miscanthi (Acari: Tetranychidae). AB - Two forms showing different male-to-male aggressiveness, different male morphologies and different diapause attributes are known in Stigmaeopsis miscanthi (Saito), a social spider mite infesting Chinese silver grass (Miscanthus sinensis Anderss). Reproductive isolation exists between the forms, although it is not always complete, and the details of their distributional patterns are unknown, but expected to be parapatric. We searched for the contact zone at Mt. Unzen (Nagasaki Pref., Japan) where both forms are known to occur at different altitudes. We found the two forms together in several M. sinensis stands, suggesting there is frequent contact between the forms in their boundary area. We discuss the mechanism(s) that maintain the parapatry related to their frequent contact and the pattern of reproductive isolation between them. PMID- 18392939 TI - Effects of temporal lobe lesions on retrograde memory: a critical review. AB - Scientific interest in retrograde memory has grown considerably over the last two decades. Studies have demonstrated that several brain regions, including the temporal lobes, participate in the formation, maintenance and storage of past memories. Different patterns and extents of retrograde memory deficits have been recorded in patients with temporal lobe lesions. It has been argued that the pattern of retrograde memory deficits is determined not only by the side and specific location of lesion within this region, but also by methodology, medical variables (such as etiology or seizure history), and coexisting cognitive deficits. In this paper we will review published case and group studies of patients with temporal lobe lesions to evaluate the impact of these factors on retrograde memory. PMID- 18392940 TI - A striking lack of genetic diversity across the wide-ranging amphibian Gastrophryne carolinensis (Anura: Microhylidae). AB - We examine phylogeographic structure across a wide-ranging microhylid frog (Gastrophryne carolinensis) using both mitochondrial (mtDNA) and nuclear (AFLP) data. Species with similar ecological characteristics such as large range size, low vagility, or existence across known biogeographic barriers, often are comprised of multiple, cryptic lineages. Surprisingly, our analyses of both portions of the genome show very little phylogeographic or population genetic structure. The family Microhylidae is one of the largest families of anurans with over 60 genera and around 400 species distributed across much of the world (Americas, Asia, Africa, and Madagascar), but very few phylogeographic studies have assessed intraspecific genetic diversity across the mitochondrial and nuclear genomes. Our results suggest that G. carolinensis, one of only three species of microhylid native to the US, has experienced a severe population bottleneck with subsequent range expansion. Comparable molecular data from closely related microhylids, in addition to demographic and ecological analyses, will provide valuable insight into patterns of genetic diversity and the processes driving phylogeographic diversity in these wide-ranging frogs. PMID- 18392942 TI - Comparative reproduction of Varroa destructor in different types of Russian and Italian honey bee combs. AB - Earlier studies showed that Russian honey bees support slow growth of varroa mite population. We studied whether or not comb type influenced varroa reproduction in both Russian and Italian honey bees, and whether Russian bees produced comb which inhibited varroa reproduction. The major differences found in this study concerned honey bee type. Overall, the Russian honey bees had lower (2.44 +/- 0.18%) levels of varroa infestation than Italian honey bees (7.20 +/- 0.60%). This decreased infestation resulted in part from a reduced number of viable female offspring per foundress in the Russian (0.85 +/- 0.04 female) compared to the Italian (1.23 +/- 0.04 females) honey bee colonies. In addition, there was an effect by the comb built by the Russian honey bee colonies that reduced varroa reproduction. When comparing combs having Russian or Italian colony origins, Russian honey bee colonies had more non-reproducing foundress mites and fewer viable female offspring in Russian honey bee comb. This difference did not occur in Italian colonies. The age of comb in this study had mixed effects. Older comb produced similar responses for six of the seven varroa infestation parameters measured. In colonies of Italian honey bees, the older comb (2001 dark) had fewer (1.13 +/- 0.07 females) viable female offspring per foundress than were found in the 2002 new (1.21 +/- 0.06 females) and 1980s new (1.36 +/- 0.08 females) combs. This difference did not occur with Russian honey bee colonies where the number of viable female offspring was low in all three types of combs. This study suggests that honey bee type largely influences growth of varroa mite population in a colony. PMID- 18392943 TI - Aerodynamic advantages of upside down take-off for aerial dispersal in Tetranychus spider mites. AB - Aerial dispersal may be important for redistribution of spider mites into new habitats. Evidence for behavioral control of aerial take-off has been well documented for Tetranychus urticae Koch. Before aerial dispersal they exhibit the aerial take-off posture that involves lifting the forelegs upright and raising the forebody. However, whether the aerial take-off posture functions to increase drag has remained unclear. The objectives of this study were to clarify: (i) aerodynamic effects of the aerial take-off posture; and (ii) actual aerial take off behavior in T. urticae. To evaluate the aerodynamic forces experienced by grounded spider mites in different postures, we constructed three-dimensional models of T. urticae, exhibiting the aerial take-off posture and the normal posture, using computer graphics. We found that the aerial take-off posture was effective in receiving greater rearward forces from wind rather than upward forces. As a result, aerial take-off from a horizontal platform is unlikely. Instead, inverted departure surfaces, e.g., lower leaf surfaces, with inclines are likely to be effective sites for take-off. Laboratory experiments and field observations indicated that the mites preferentially adopted such a position for orientation and take-off. Our findings provided a rationale for the take-off behavior of Tetranychus spider mites. PMID- 18392944 TI - Influence of body mass index on prognosis in gynecological malignancies. AB - OBJECTIVE: To evaluate the effect of obesity on the presentation and course of disease in patients with gynecological cancers. STUDY DESIGN: Records of patients with endometrial (n = 1180), cervical (n = 738), and ovarian cancer (n = 824) treated between 1986 and 2005 were reviewed. Body mass index (BMI) was analyzed in relation to tumor stage, tumor grading, and prognosis. Steroid hormone receptor status and growth fraction (MIB1; Ki-67-antigen) of tumors in relation to BMI were analyzed in subgroups with endometrial (n = 183) and advanced ovarian (n = 221) cancers. In the latter subgroup, tumor vascularization (CD31) and expression of bcl-2, c-erb-B2, fibronectin, and tumor markers (CA-125, CA15-3, CEA) were also evaluated. Statistical analyses included bivariate correlation, cross-tabulation, Kaplan-Meier-survival analyses, and multifactorial residual survival analyses. RESULTS: Obese patients with endometrial carcinoma were significantly younger (p < 0.001) and their tumors were less advanced at diagnosis (p = 0.001) and were better differentiated (p = 0.010). In the subgroups, neither steroid hormone receptor status nor MIB1-determined growth fraction correlated with BMI. For both endometrial and cervical carcinomas, a high BMI influenced overall survival favorably (p (endometrial )= 0.004 and p (cervical )= 0.026). In ovarian cancer, there was a trend toward improved survival in more obese patients (p = 0.053). Immunohistochemistry revealed that c erb-B2 expression was slightly lower in tumors of obese patients (r = -0.142; p = 0.039), but BMI did not influence any other factor. CONCLUSIONS: Although obesity increases the incidence of cancer, a high BMI does not seem to adversely influence the prognosis in patients with the mentioned gynecological malignancies. PMID- 18392945 TI - A prospective analysis of testicular androgenic function in recipients of a renal allograft. AB - Eighteen adult males with end stage renal disease (ESRD) were studied to determine the serum levels of gonadotropins (LH and FSH), prolactin (PRL) and testosterone. All of the patients were studied longitudinally while undergoing maintenance hemodialysis (HD) and six months after renal transplantation. Prior to transplantation, significantly high levels of gonadotropins and PRL were observed. During HD the serum testosterone levels tended to be subnormal in most of the uremic patients and low normal in some of the subjects. Renal transplantation led to a significant improvement (P < 0.05) in serum testosterone. Elevated gonadotropin and PRL levels observed in patients on HD returned to the normal range in most of the patients after successful renal transplantation. PMID- 18392947 TI - Common morality: comment on Beauchamp and Childress. AB - The notion of common morality plays a prominent role in some of the most influential theories of biomedical ethics. Here, I focus on Beauchamp and Childress's models in the fourth and fifth edition of Principles of Biomedical Ethics as well as on a revision that Beauchamp proposed in a recent article. Although there are significant differences in these works that require separate analysis, all include a role for common morality as starting point and normative framework for theory construction in combination with a coherence theory of moral justification. I defend to some extent the existence and empirical significance of common morality, as delineated by Beauchamp and Childress in different versions, but criticize its normative role. It is neither convincing as a moral foundation nor well compatible with a standard coherentist justification. I suggest that the authors should give up the foundational account for a more modest account of common morality as resource of well-established moral insights and experiences, which have proved generally valid but neither sufficient nor infallible. Beauchamp's latest proposal appears as a step in this direction; indeed, it may be the beginning of the end of his common-morality theory. PMID- 18392946 TI - Matrine induces programmed cell death and regulates expression of relevant genes based on PCR array analysis in C6 glioma cells. AB - Matrine, one of the main components extracted from Sophora flavescens Ait, has a wide range of pharmacological effects including anti-tumor activities on a number of cancer cell lines. This study has investigated whether matrine could also display anti-tumor action on rat C6 glioma cells. Exposure of C6 cells to matrine resulted in inhibition of proliferation and induction of apoptosis in a dose dependent manner, as measured by the MTT assay and Flow cytometry. The Annexin V/PI staining further detected the apoptotic cells at both early and late phases of apoptosis. We used AO/EB staining to examine the programmed cell death of matrine-treated C6 cells, and showed that the death rate detected by AO/EB staining was higher than the apoptosis rate measured by Annexin V/PI staining, suggesting that autophagy, the Type II programmed cell death, may be involved in matrine-induced cell death, which was further confirmed by electronic microscopy. To explore the molecular mechanism, an apoptosis real-time PCR array was performed, which has demonstrated that 57 genes were at least 2-fold upregulated, and 11 genes were at least 2-fold downregulated in matrine-treated C6 cells, compared with untreated cells. However, the gene expression profiles could only partly and roughly explain molecular mechanisms of apoptosis and autophagy in matrine-treated C6 cells, thus further investigations are required to confirm the specific molecular pathways and related molecules responsible for the programmed cell death. PMID- 18392948 TI - Turning failures into successes: a methodological shortcoming in empirical research on surrogate accuracy. AB - Decision making for incompetent patients is a much-discussed topic in bioethics. According to one influential decision making standard, the substituted judgment standard, a surrogate decision maker ought to make the decision that the incompetent patient would have made, had he or she been competent. Empirical research has been conducted in order to find out whether surrogate decision makers are sufficiently good at doing their job, as this is defined by the substituted judgment standard. This research investigates to what extent surrogates are able to predict what the patient would have preferred in the relevant circumstances. In this paper we address a methodological shortcoming evident in a significant number of studies. The mistake consists in categorizing responses that only express uncertainty as predictions that the patient would be positive to treatment, on the grounds that the clinical default is to provide treatment unless it is refused. We argue that this practice is based on confusion and that it risks damaging the research on surrogate accuracy. PMID- 18392949 TI - Justifying group-specific common morality. AB - Some defenders of the view that there is a common morality have conceived such morality as being universal, in the sense of extending across all cultures and times. Those who deny the existence of such a common morality often argue that the universality claim is implausible. Defense of common morality must take account of the distinction between descriptive and normative claims that there is a common morality. This essay considers these claims separately and identifies the nature of the arguments for each claim. It argues that the claim that there is a universal common morality in the descriptive sense has not been successfully defended to date. It maintains that the claim that there is a common morality in the normative sense need not be understood as universalist. This paper advocates the concept of group specific common morality, including country-specific versions. It suggests that both the descriptive and the normative claims that there are country-specific common moralities are plausible, and that a country specific normative common morality could provide the basis for a country's bioethics. PMID- 18392950 TI - Release of heat shock protein 70 and the effects of extracellular heat shock protein 70 on the production of IL-10 in fibroblast-like synoviocytes. AB - It has recently been suggested that heat shock protein (Hsp) 70, an intracellular protein, can be released into the extracellular compartment and exert important immunomodulatory functions. Although elevated Hsp70 has been found in synovial fluid from patients with rheumatoid arthritis (RA), its sources and extracellular functions remain unclear. In this study, we explored whether stress response such as heat stress or exposure to tumor necrosis factor-alpha (TNF-alpha) could induce Hsp70 release from RA fibroblast-like synoviocytes (FLSs) and whether extracellular Hsp70 would stimulate cytokine production in RA FLSs. Cultured FLSs were obtained from patients with RA. The expression of intracellular Hsp70 was studied by Western blot. Hsp70 release and the production of interleukin (IL)-6, IL-8, and IL-10 by RA FLSs were studied by specific enzyme-linked immunosorbent assays. The levels of Toll-like receptor (TLR) 2 and 4 mRNA and protein in FLSs were analyzed using reverse transcription-polymerase chain reaction and Western blotting. Treatment with sublethal heat shock or TNF-alpha results in the up regulation of intracellular Hsp70 in FLSs and Hsp70 release from RA FLSs. In vitro studies show that extracellular Hsp70 can induce anti-inflammatory cytokine IL-10 production in FLSs. The mRNA and protein expression of TLR2 and TLR4 was demonstrated in FLSs, and TLR4 blocking abrogated the up-regulatory effects of Hsp70 on IL-10 production. Thus, these results lend support to the hypothesis that Hsp70 is actively released from FLSs in response to heat shock or TNF-alpha and Hsp70 may be a major paracrine/autocrine inducer of IL-10 production in FLSs via TLR4. PMID- 18392951 TI - Bicyclol: a novel antihepatitis drug with hepatic heat shock protein 27/70 inducing activity and cytoprotective effects in mice. AB - Heat shock proteins (HSPs) are the best-known endogenous factors that protect against cell injury under various pathological conditions and that can be induced by various physical, chemical, and biological stressors. New research seeks to discover a compound that is clinically safe and can induce the accumulation of HSPs in patients. This paper reports that the oral administration of three doses of bicyclol, a novel antihepatitis drug, induced hepatic HSP27 and HSP70 expression in a time- and dose-dependent manner, and that bicyclol treatment stimulated heat shock factor 1 (HSF1) activation in mice. The inducing effects of bicyclol on HSP27, HSP70 and HSF1 were all blocked by quercetin, an inhibitor of HSP biosynthesis. The cytoprotective effect of HSP27/70 induced by bicyclol against hepatotoxicity of acetaminophen (AP) was assessed in mice. The prior administration of bicyclol markedly suppressed AP-induced liver injury as indicated by the reduction in the elevation of serum alanine aminotransferase and aspartate aminotransferase, in liver necrosis, in the release of cytochrome c and apoptosis-inducing factor from mitochondria, as well as in hepatic deoxyribonucleic acid fragmentation in mice. However, all the above actions of bicyclol against AP-induced mouse liver injuries were significantly attenuated by quercetin. This is the first report to show that bicyclol induces hepatic HSP27/70 expression via activation of HSF1 and that the cytoprotective action of bicyclol against liver injury is mediated by its induction of HSP27/70. These results provide new evidence for elucidating the mechanism of the hepatoprotective action of bicyclol in animals and patients. PMID- 18392952 TI - Monitoring of cytomegalovirus reactivation in bone marrow transplant recipients by real-time PCR. AB - Cytomegalovirus (CMV) has been recognized as the most important viral pathogen in persons undergoing bone marrow transplantation (BMT). The aim was to develop a quantitative PCR assay to quantify CMV DNA in peripheral blood leukocytes (PBLs) of bone marrow transplantation (BMT) patients. An in-house real-time PCR assay based on TaqMan technology was developed to monitor the quantity of CMV DNA in PBLs of the BMT recipients. Sequential blood samples (415 specimens) were collected from 43 patients as weekly intervals until day 100 after transplantation. The CMV DNA was quantified in parallel with the pp65 antigenemia assay in PBL samples. Viral reactivation occurred in 51% and 41.8% of the recipients as detected by RQ-PCR and antigenemia assays respectively. There was a significant correlation between both assays (P < 0.0001); however, the RQ-PCR was more sensitive than the antigenemia. CMV DNA was detected by the RQ-PCR by a median of 14 days earlier than the antigenemia. Preemptive therapy was implemented in the antigenemia positive cases. The administration of ganciclovir led to a rapid decrease in the viral load. After preemptive therapy, the antigenemia achieved a negative result earlier than the RQ-PCR assay (a median of 17.5 days). An increase of viral load in both quantitative assays and of cyclosporine serum level were identified as the most significant risk factors for CMV reactivation. The quantitative CMV PCR might be a useful tool for monitoring the CMV reactivation and guiding the efficacy of the CMV preemptive therapy in BMT recipients. PMID- 18392954 TI - Clinical, histopathological, immunophenotypic and molecular analysis of 60 patients with cutaneous T-cell infiltrates with follow up of indeterminate cases to identify T-cell lymphoma. AB - Diagnosis of primary cutaneous T-cell lymphomas, especially of mycosis fungoides could be difficult in early stage due to clinical and histopathological similarity to reactive inflammatory dermatoses. To assess diagnostic value of complex histological, immunophenotypic and T-cell receptor gamma gene rearrangement analysis, skin biopsy specimen and peripheral blood samples of 60 patients with suspected cutaneous T-cell lymphoma were analyzed. Our results indicate clear distinction between reactive dermatoses (benign cases, n = 31) and cutaneous T-cell lymphomas (lymphoma cases, n = 17). As definite diagnosis was not obtained in a smaller group of patients (indeterminate cases, n = 12), these patients were followed up. Repeated skin biopsy confirmed mycosis fungoides in 6/12 cases, however in 6/12 patients the diagnosis remained indeterminate. We concluded that careful and complex clinical follow up and repeated histopathological, immunophenotypic and molecular analysis is needed for an appropriate diagnosis in the assessment of early stage mycosis fungoides and uncertain clinical cases. PMID- 18392953 TI - Alterations of the CD4(+), CD8 (+) T cell subsets, interleukins-1beta, IL-10, IL 17, tumor necrosis factor-alpha and soluble intercellular adhesion molecule-1 in rheumatoid arthritis and osteoarthritis: preliminary observations. AB - Rheumatoid arthritis is a multisystem disease with underlying immune mechanisms. Osteoarthritis is a debilitating, progressive disease of diarthrodial joints associated with the aging process. Although much is known about the pathogenesis of rheumatoid arthritis and osteoarthritis, our understanding of some immunologic changes remains incomplete. This study tries to examine the numeric changes in the T cell subsets and the alterations in the levels of some cytokines and adhesion molecules in these lesions. To accomplish this goal, peripheral blood and synovial fluid samples were obtained from 24 patients with rheumatoid arthritis, 15 patients with osteoarthritis and six healthy controls. The counts of CD4 (+) and CD8 (+) T lymphocytes were examined using flow cytometry. The levels of some cytokines (TNF-alpha, IL1-beta, IL-10, and IL-17) and a soluble intercellular adhesion molecule-1 (sICAM-1) were measured in the sera and synovial fluids using enzyme linked immunosorbant assay. We found some variations in the counts of T cell subsets, the levels of cytokines and sICAM-1 adhesion molecule between the healthy controls and the patients with arthritis. High levels of IL-1beta, IL-10, IL-17 and TNF-alpha (in the serum and synovial fluid) were observed in arthritis compared to the healthy controls. In rheumatoid arthritis, a high serum level of sICAM-1 was found compared to its level in the synovial fluid. A high CD4(+)/CD8(+) T cell ratio was found in the blood of the patients with rheumatoid arthritis. In rheumatoid arthritis, the cytokine levels correlated positively with some clinicopathologic features. To conclude, the development of rheumatoid arthritis and osteoarthritis is associated with alteration of the levels of some cytokines. The assessment of these immunologic changes may have potential prognostic roles. PMID- 18392955 TI - Intrapulmonary rheumatoid nodules in a patient with long-standing rheumatoid arthritis treated with leflunomide. AB - Rheumatoid nodules are well established manifestations of rheumatoid arthritis but in the lungs they are very rare according to the literature. In our study we present the case of a 34-year-old woman with rheumatoid arthritis and secondary Sjogren's syndrome who developed multiplex rheumatoid nodules in the lungs 3 years after initiating leflunomide therapy. During leflunomide therapy we did not detect inflammation in the joints. Surprisingly, in November 2005 she started to cough, had low grade fever and low back pain. On the chest X-ray there were multiplex necrobiotic nodules in the lungs. All bacteriological, viral and fungal investigations including tuberculosis, serological tests and cytology were negative. The X-ray, video-associated thoracoscopy and repeated biopsy of the lung followed by histology of the samples proved intrapulmonary rheumatoid nodules, caused by leflunomide. PMID- 18392956 TI - A pilot study for human tumor/DNA banking: returned more questions than answers. AB - A pilot study was performed for setting up the Dokuz Eylul University Breast Tumor DNA Bank (DEUBTB) to facilitate the sharing of tumor DNA/RNA samples and related data from cases collected by collaborators specializing in the breast cancer diseases between 2004 and 2006. The pilot study aimed to provide answers for certain questions on: (1) ethical concerns (informing the volunteer for donating specimen, anonymizing the sample information, procedure on sample request), (2) obtaining and processing samples (technical issues, flowchart), (3) storing samples and their products (storing forms and conditions), (4) clinical database (which clinical data to store), (5) management organization (quality and quantity of personnel, flowchart for management relations), (6) financial issues (establishment and maintenance costs). When the bank had 64 samples, even though it is quite ready to supply samples for a research project, it revealed many questions on details that may be answered in more than one way, pointing that all biobanks need to be controlled by a higher degree of management party which develops and offers quality standards for these establishments. PMID- 18392957 TI - Overview of whole-genome essentiality analysis. PMID- 18392958 TI - Transposon-based strategies for the identification of essential bacterial genes. AB - We present a conceptual review of transposition-based strategies for determining gene essentiality on a one-by-one basis in bacteria. Many of the techniques are described in greater detail in individual chapters of this volume. The second section of this chapter deals with transposition-deletion-based strategies for determining the essentiality of blocks of genes. This latter approach has the potential to experimentally define the minimal required genome for a given organism. PMID- 18392959 TI - Identification and analysis of essential genes in Haemophilus influenzae. AB - The human respiratory pathogen Haemophilus influenzae, a Gram-negative bacterium, is the first free-living organism to have its complete genome sequenced, providing the opportunity to apply genomic-scale approaches to study gene function. This chapter provides an overview of a highly efficient, in vitro mariner transposon-based method that exploits the natural transformation feature of this organism for the identification of essential genes. In addition, we describe strategies for conditional expression systems that would facilitate further analysis of this class of genes. Finally, we outline a method based on the approach used in H. influenzae for identifying essential genes that can be applied to other bacteria that are not naturally transformable. PMID- 18392960 TI - Transposon site hybridization in Mycobacterium tuberculosis. AB - Microarray mapping of transposon insertions can be used to quantify the relative abundance of different transposon mutants within a complex pool after exposure to selective pressure. The transposon site hybridization (TraSH) method applies this strategy to the study of Mycobacterium tuberculosis and can be adapted to the study of other microorganisms. This chapter describes the methods used to mutagenize mycobacteria with transposons, extract genomic DNA, amplify genomic DNA adjacent to transposon ends using polymerase chain reaction and T7 transcription, and synthesize labeled cDNA. It also describes methods used to construct an appropriate microarray, hybridize labeled cDNA, and analyze the microarray data. Important considerations involved in the experimental design of the selective pressure, the design of the microarray, and the statistical analysis of collected data are discussed. PMID- 18392961 TI - Essential genes in the infection model of Pseudomonas aeruginosa PCR-based signature-tagged mutagenesis. AB - PCR-based signature tagged mutagenesis is an "en masse" screening technique based upon unique oligonucleotide tags (molecular barcodes) for identification of genes that will diminish or enhance maintenance of an organism in a specific ecological niche or environment. PCR-based STM applied to Pseudomonas aeruginosa permitted the identification of genes essential or in vivo maintenance by transposon insertion and negative selection in a mixed population of bacterial mutants. The innovative adaptations and refinement of the technology presented here with P. aeruginosa STM mutants selected in the rat lung have given critical information about genes essential for causing a chronic infection and a wealth of information about biological processes in vivo. The additional use of competitive index analysis for measurement of the level of virulence in vivo, microarray-based screening of selected prioritized STM mutants coupled to metabolomics analysis can now be attempted systematically on a genomic scale. PCR-based STM and combined whole-genome methods can also be applied to any organism having selectable phenotypes for screening. PMID- 18392962 TI - Whole-genome detection of conditionally essential and dispensable genes in Escherichia coli via genetic footprinting. AB - We present a whole-genome approach to genetic footprinting in Escherichia coli using Tn5-based transposons to determine gene essentiality. A population of cells is mutagenized and subjected to outgrowth under selective conditions. Transposon insertions in the surviving mutants are detected using nested polymerase chain reaction (PCR), agarose gel electrophoresis, and software-assisted PCR product size determination. Genomic addresses of these inserts are then mapped onto the E. coli genome sequence based on the PCR product lengths and the addresses of the corresponding genome-specific primers. Gene essentiality conclusions were drawn based on a semiautomatic analysis of the number and relative positions of inserts retained within each gene after selective outgrowth. PMID- 18392963 TI - Generating a collection of insertion mutations in the Staphylococcus aureus genome using bursa aurealis. AB - Staphylococcus aureus is the leading cause of wound and hospital-acquired infections. The emergence of strains with resistance to all antibiotics has created a serious public health problem. Transposon-based mutagenesis can be used to generate libraries of mutants and to query genomes for factors involved in nonessential pathways, such as virulence and antibiotic resistance. Ideally, such studies should employ defined and complete sets of isogenic mutants and should be conducted so as to permit acquisition and comparison of the complete data sets. Such systematic knowledge can reveal entire pathways and can be exploited for the rational design of therapies. The mariner-based transposon, bursa aurealis, can be used to generate random libraries of mutants in laboratory strains and clinical isolates of S. aureus. This chapter describes a procedure for isolating mutants and mapping the insertion sites on the chromosome. PMID- 18392964 TI - Multipurpose transposon insertion libraries for large-scale analysis of gene function in yeast. AB - Transposons have long been recognized as useful laboratory tools facilitating genome-scale studies of gene function. Relative to traditional methods, transposon mutagenesis offers a rapid and economical means of generating large numbers of independent insertions in target DNA through minimal experimental manipulation. In particular, the transposon insertion library described here is an excellent tool for the analysis of gene function on a large scale in the budding yeast Saccharomyces cerevisiae. The transposon utilized in this library is multifunctional, such that the library can be used to screen for disruption phenotypes while also providing a means to generate epitope-tagged alleles and, in many cases, conditional alleles. Provided here are complete protocols by which the transposon insertion library may be used to screen for mutant phenotypes in yeast as well as accompanying protocols describing a means of identifying transposon insertion sites within strains of interest. In total, this insertion library is a singularly useful tool for genome-wide functional analysis, and the general approach is applicable to other organisms in which transforming DNA tends to integrate by homologous recombination. PMID- 18392965 TI - How to make a defined near-saturation mutant library. Case 1: Pseudomonas aeruginosa PAO1. AB - We have constructed a near-saturation level mutant library for Pseudomonas aeruginosa strain PAO1 using Tn5-derived transposons mapped to the PAO1 reference sequence. This chapter describes the high-throughput techniques used to generate and map the mutant strains. In addition, an analysis of the utility of this collection is presented based on changes to the annotation for the PAO1 genome in the past years, as well as the citation record for this collection. It is clear that many avenues of research have been accelerated by this collection and that additional large mutant strain collections will further aid in defining gene function and biological processes in pathogens. PMID- 18392966 TI - Comparing insertion libraries in two Pseudomonas aeruginosa strains to assess gene essentiality. AB - Putative essential genes can be identified by comparing orthologs not disrupted in multiple near-saturated transposon insertion mutation libraries in related strains of the same bacterial species. Methods for identifying all orthologs between two bacterial strains and putative essential orthologs are described. In addition, protocols detailing near-saturation transposon insertion mutagenesis of bacteria are presented, including (1) conjugation-mediated mutagenesis, (2) automated colony picking and liquid handling of mutant cultures, and (3) arbitrary polymerase chain reaction amplification and sequencing of genomic DNA adjacent to transposon insertion sites. PMID- 18392967 TI - The construction of systematic in-frame, single-gene knockout mutant collection in Escherichia coli K-12. AB - Here we describe the systematic construction of well-defined, in-frame, single gene deletions of all nonessential genes in Escherichia coli K-12. The principal strategy is based on the method for one-step inactivation of chromosomal genes in E. coli K-12 established by Datsenko and Wanner (1), namely, the replacement of a target gene with a selectable antibiotic-resistant marker generated by polymerase chain reaction (PCR) using oligonucleotide DNA primers homologous to the gene flanking regions. The advantages of this method include complete deletion of an entire open reading frame and precise design eliminating polar effects for the downstream genes on E. coli chromosome. PMID- 18392968 TI - The applications of systematic in-frame, single-gene knockout mutant collection of Escherichia coli K-12. AB - The increasing genome sequence data of microorganisms has provided the basis for comprehensive understanding of organisms at the molecular level. Besides sequence data, a large number of experimental and computational resources are required for genome-scale analyses. Escherichia coli K-12 has been one of the best characterized organisms in molecular biology. Recently, the whole-genome sequences of two closely related E. coli K-12 strains, MG1655 (1) and W3110 (2), were compared and confirmed by resequencing selected regions from both strains (2). The availability of highly accurate E. coli K-12 genomes provided an impetus for the cooperative reannotation of both MG1655 and W3110 (3). A set of precisely defined, single-gene knockout mutants of all nonessential genes in E. coli K-12 was constructed based on the recent accurate genome sequence data ([4] and Chapter 11). These mutants were designed to create in-frame (nonpolar) deletions upon elimination of the resistance cassette. These mutants have provided new key information on E. coli biology. First, the vast majority of the 3985 genes that were independently disrupted at least twice are probably nonessential, at least under the conditions of selection. Second, the 303 genes that we repeatedly failed to disrupt are candidates for E. coli essential genes. Lastly, phenotypic effects of all these mutations in the uniform genetic background of E. coli BW25113 were assessed by profiling mutants' growth yields on rich and minimal media (4). These mutants should provide not only a basic resource for systematic functional genomics but also an experimental data source for systems biology applications. The mutants can serve as fundamental tools for a number of reverse genetics approaches, permitting analysis of the consequences of the complete loss of gene function, in contrast with forward genetics approaches in which mutant phenotypes are associated with a corresponding gene or genes. PMID- 18392969 TI - A novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12. AB - We developed a novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12. The basic idea of the method is to randomly disrupt the genes on the DNA fragments cloned on the Kohara library by inserting a mini-transposon first, and then transfer the disrupted genes from the lambda vector to the E. coli chromosome by homologous recombination. Using this method, we constructed a set of 8402 Km(r) cis-diploid mutants harboring a mini-Tn10 insertion mutation and the corresponding wild-type gene on a chromosome, as well as a set of 6954 haploid mutants derived from the cis-diploid mutants. The major advantage of the strategy used is that the indispensable genes or sites for growth can be identified. Preliminary results suggest that 415 open reading frames are indispensable for growth in E. coli cells. A total of 6404 haploid mutants were deposited to Genetic Strains Research Center, National Institute of Genetics, Japan (Chapter 26) and are available for public distribution upon request (http://shigen.lab.nig.ac.jp/ecoli/strain/nbrp/resource.jsp). PMID- 18392970 TI - High-throughput creation of a whole-genome collection of yeast knockout strains. AB - Gene disruption methods have proved to be a valuable tool for studying gene function in yeast. Gene replacement with a drug-resistant cassette renders the disruption strain selectable and is stable against reversion. Polymerase chain reaction-generated deletion cassettes are designed with homology sequences that flank the target gene. These deletion cassettes also contain unique "molecular bar code" sequence tags. Methods to generate these mutant strains are scalable and facile, allowing for the production of a collection of systematic disruptions across the Saccharomyces cerevisiae genome. The deletion strains can be studied individually or pooled together and assayed in parallel utilizing the sequence tags with microarray-based methods. PMID- 18392971 TI - Analysis of genetic interactions on a genome-wide scale in budding yeast: diploid based synthetic lethality analysis by microarray. AB - Comprehensive collections of open reading frame (ORF) deletion mutant strains exist for the budding yeast Saccharomyces cerevisiae. With great prescience, these strains were designed with short molecular bar codes or TAGs that uniquely mark each deletion allele, flanked by shared priming sequences. These features have enabled researchers to handle yeast mutant collections as complex pools of approximately 6000 strains. The presence of any individual mutant within a pool can be assessed indirectly by measuring the relative abundance of its corresponding TAG(s) in genomic DNA prepared from the pool. This is readily accomplished by wholesale polymerase chain reaction (PCR) amplification of the TAGs using fluorescent oligonucleotide primers that recognize the common flanking sequences, followed by hybridization of the labeled PCR products to a TAG oligonucleotide microarray. Here we describe a method-diploid-based synthetic lethality analysis by microarray (dSLAM)-whereby such pools can be manipulated to rapidly construct and assess the fitness of 6000 double-mutant strains in a single experiment. Analysis of double-mutant strains is of growing importance in defining the spectrum of essential cellular functionalities and in understanding how these functionalities interrelate. PMID- 18392972 TI - Scarless engineering of the Escherichia coli genome. AB - E. coli K-12, being one of the best understood and thoroughly analyzed organisms, is the workhorse of genetic, biochemical, and systems biology research, as well as the platform of choice for numerous biotechnological applications. Genome minimization/remodeling is now a feasible approach to further enhance its beneficial characteristics for practical applications. Two genome engineering techniques, a lambda Red-mediated deletion method and a suicide (conditionally replicative) plasmid-based allele replacement procedure are presented here. These techniques utilize homologous recombination, and allow the rapid introduction of virtually any modifications in the genome. PMID- 18392973 TI - Minimization of the Escherichia coli genome using the Tn5-targeted Cre/loxP excision system. AB - Efficient genome-engineering tools have been developed for use in whole-genome essentiality studies. In this chapter, we describe a powerful genomic deletion tool, the Tn5-targeted Cre/loxP excision system, for determining genetic essentiality and minimizing bacterial genomes on a genome-wide scale. This tool is based on the Tn5 transposition system, phage P1 transduction, and the Cre/loxP excision system. We have generated two large pools of independent transposon insertion mutants in Escherichia coli using random transposition of two modified Tn5 transposons (TnKloxP and TnCloxP) with two different selection markers, kanamycin-resistance gene (Km(R)) or chloramphenicol-resistance gene (Cm(R)), and a loxP site. Transposon integration sites are identified by direct genome sequencing of the genomic DNA. By combining a mapped transposon mutation from each of the mutant pools into the same chromosome using phage P1 transduction and then excising the nonessential genomic regions flanked by the two loxP sites using Cre-mediated loxP recombination, we can obtain numerous E. coli deletion strains from which nonessential regions of the genome are deleted. In addition to the combinatorial deletion of the E. coli genomic regions, we can create a cumulative E. coli deletion strain from which all the individual deleted regions are excised. This process will eventually yield an E. coli strain in which the genome is reduced in size and contains only regions that are essential for viability. PMID- 18392974 TI - Construction of long chromosomal deletion mutants of Escherichia coli and minimization of the genome. AB - Genetic information consists of protein- and RNA-coding genes that exist in a range of sizes and noncoding cis- and trans-acting sequence elements. The use of long chromosomal deletion mutations is a powerful method for identifying essential genetic information through experimental reduction of the genome to its minimal gene set. Taking advantage of recent technical advances, we constructed sequence-specific long deletion mutations of the Escherichia coli chromosome. In a recent report (1), we described a set of E. coli medium-scale deletions (MDs) and large-scale deletions (LDs). Several LD mutations were combined to generate an engineered strain lacking approximately 30% of the parental chromosome. We then constructed another set of deletion mutations, MDs and small-scale deletions (SDs), and identified additional essential genetic regions using complementation analysis. To delete the remaining essential chromosomal regions, we developed an Flp recombinase target (FRT)-based system of site-specific recombination to move chromosomal regions onto mini-F plasmids in vivo. In this report, we describe the details of the construction of several of these types of large chromosomal deletion mutants. PMID- 18392975 TI - Identification of essential genes in Staphylococcus aureus by construction and screening of conditional mutant library. AB - Antisense RNA technology has been used effectively to downregulate gene expression in a variety of bacterial systems. Regulated antisense RNA strategy provides an important approach to identify and characterize essential genes critical to bacterial growth in vitro and in vivo. This strategy allows selective genes to be turned on or off and to be expressed at certain levels. The availability of the Staphylococcus aureus (S. aureus) genome sequence makes it feasible to generate a gene-specific antisense RNA library. The combination of regulated antisense RNA technology and the gene-specific antisense RNA library allows for genome-wide analyses of functions of staphylococcal gene products for growth in culture and survival during infection. PMID- 18392976 TI - Techniques for the isolation and use of conditionally expressed antisense RNA to achieve essential gene knockdowns in Staphylococcus aureus. AB - This chapter provides methods and insights into the use of antisense RNA as a molecular genetic tool. Posttranscriptional inhibition of specific gene expression can be achieved by antisense RNA fragments under control of a conditional promoter. Effective titration of gene expression can cause an apparent null mutation or can be modulated to levels of interest in comparison with wild type. Validation of antisense RNA can be achieved by both RNA and protein quantitation techniques. Applications include phenotypic studies of genes in response to specific stimuli, environments, or the contribution of genes in regulatory networks. This chapter will focus on shotgun-cloned antisense for comprehensive gene identification and cell-based hypersensitivity assays for antibiotic screening. Antisense RNA strategies have high utility when the target gene is essential for survival and needs to be compared with wild type. PMID- 18392977 TI - Introduction of conditional lethal amber mutations in Escherichia coli. AB - A method is described for generating conditional lethal mutations in essential genes in Escherichia coli. In this procedure, amber stop codons are introduced as "tagalong" mutations in the flanking DNA of a downstream antibiotic-resistance marker by lambda Red recombination. The marker is removed by expression of I-SceI homing endonuclease, leaving a markerless mutation. The mutants then depend upon expression of a suppressor transfer RNA (tRNA) for survival, which is expressed under control of the arabinose promoter on a high-copy-number plasmid. PMID- 18392978 TI - Statistical methods for building random transposon mutagenesis libraries. AB - During the construction of random transposon mutagenesis libraries, four essential statistical issues arise: (1) Computing basic probability results for number of open reading frame knockouts. (2) Estimating the number of new open reading frames that will be knockouts in the next set of clones. (3) Estimating the number of essential open reading frames. (4) Computing the probability that an open reading frame is essential given the distribution of insertions. This chapter examines these issues and evaluates potential solutions using three different approaches: Efron and Thisted's estimator, Will and Jacobs's parametric bootstrap, and Blades and Broman's Gibbs sampler. In doing so, this chapter provides guidance for using the R statistical project to solve these problems. PMID- 18392979 TI - Statistical evaluation of genetic footprinting data. AB - As transposomics is extended to genome scale, appropriate statistical methods need to be developed to assign significance to gene essentiality. In this chapter, the author presents a set of steps that, together with genome-scale insertion data and the complete genome sequence of a prokaryote, can be used to classify the genes of the organism as either "essential" or "nonessential." PMID- 18392980 TI - Modeling competitive outgrowth of mutant populations: why do essentiality screens yield divergent results? AB - Mutant propagation (outgrowth) is an important step in all large-scale gene essentiality experiments, profoundly influencing essentiality assignment produced. Using a simplified mathematical model of competitive outgrowth in a diverse mutant population, we have identified several technological factors (duration of outgrowth, sensitivity of the scoring technique, initial cell titer of each mutant in the population) that have the largest impact on the outcome of the essentiality screen. The model can be used for planning a large-scale gene essentiality screen as well as for analyzing its results, including meaningful comparisons of "essential" gene lists generated by different techniques. PMID- 18392981 TI - Statistical analysis of fitness data determined by TAG hybridization on microarrays. AB - TAG, or bar-code, microarrays allow measurement of the oligonucleotide sequences (TAGs) that mark each strain of deletion mutants in the Saccharomyces cerevisiae yeast knockout (YKO) collection. Comparison of genomic DNA from pooled YKO samples allows estimation of relative abundance of TAGs marking each deletion strain. Features of TAG hybridizations create unique challenges for analysis. Analysis is complicated by the presence of two TAGs in most YKO strains and the hybridization behavior of TAGs that may differ in sequence from array probes. The oligonucleotide size of labeled TAGs also results in difficulty with contaminating sequences that cause reduced specificity. We present methods for analysis that approach these unique features of TAG hybridizations. PMID- 18392982 TI - Profiling of Escherichia coli Chromosome database. AB - The Profiling of Escherichia coli Chromosome (PEC) database (http://www.shigen.nig.ac.jp/ecoli/pec/) is designed to allow E. coli researchers to efficiently access information from functional genomics studies. The database contains two principal types of data: gene essentiality and a large collection of E. coli genetic research resources. The essentiality data are based on data compilation from published single-gene essentiality studies and on cell growth studies of large-deletion mutants. Using the circular and linear viewers for both whole genomes and the minimal genome, users can not only gain an overview of the genome structure but also retrieve information on contigs, gene products, mutants, deletions, and so forth. In particular, genome-wide exhaustive mutants are an essential resource for studying E. coli gene functions. Although the genomic database was constructed independently from the genetic resources database, users may seamlessly access both types of data. In addition to these data, the PEC database also provides a summary of homologous genes of other bacterial genomes and of protein structure information, with a comprehensive interface. The PEC is thus a convenient and useful platform for contemporary E. coli researchers. PMID- 18392983 TI - Gene essentiality analysis based on DEG, a database of essential genes. AB - Essential genes are the genes that are indispensable for the survival of an organism. The genome-scale identification of essential genes has been performed in various organisms, and we consequently constructed DEG, a Database that contains currently available essential genes. Here we analyzed functional distributions of essential genes in DEG, and found that some essential-gene functions are even conserved between the prokaryote (bacteria) and the eukaryote (yeast), e.g., genes involved in information storage and processing are overrepresented, whereas those involved in metabolism are underrepresented in essential genes compared with non-essential ones. In bacteria, species specificity in functional distribution of essential genes is mainly due to those involved in cellular processes. Furthermore, within the category of information storage and processing, function of translation, ribosomal structure, and biogenesis are predominant in essential genes. Finally, some potential pitfalls for analyzing gene essentiality based on DEG are discussed. PMID- 18392984 TI - Detection of essential genes in Streptococcus pneumoniae using bioinformatics and allelic replacement mutagenesis. AB - Although the emergence and spread of antimicrobial resistance in major bacterial pathogens for the past decades poses a growing challenge to public health, discovery of novel antimicrobial agents from natural products or modification of existing antibiotics cannot circumvent the problem of antimicrobial resistance. The recent development of bacterial genomics and the availability of genome sequences allow the identification of potentially novel antimicrobial agents. The cellular targets of new antimicrobial agents must be essential for the growth, replication, or survival of the bacterium. Conserved genes among different bacterial genomes often turn out to be essential (1, 2). Thus, the combination of comparative genomics and the gene knock-out procedure can provide effective ways to identify the essential genes of bacterial pathogens (3). Identification of essential genes in bacteria may be utilized for the development of new antimicrobial agents because common essential genes in diverse pathogens could constitute novel targets for broad-spectrum antimicrobial agents. PMID- 18392985 TI - Design and application of genome-scale reconstructed metabolic models. AB - In this chapter, the process for the reconstruction of genome-scale metabolic networks is described, and some of the main applications of such models are illustrated. The reconstruction process can be viewed as an iterative process where information obtained from several sources is combined to construct a preliminary set of reactions and constraints. This involves steps such as genome annotation; identification of the reactions from the annotated genome sequence and available literature; determination of the reaction stoichiometry; definition of compartmentation and assignment of localization; determination of the biomass composition; measurement, calculation, or fitting of energy requirements; and definition of additional constraints. The reaction and constraint sets, after debugging, may be integrated into a stoichiometric model that can be used for simulation using tools such as Flux Balance Analysis (Section 3.8). From the flux distributions obtained, physiologic parameters such as growth yields or minimal medium components can be calculated, and their distance from similar experimental data provides a basis from where the model may need to be improved. PMID- 18392986 TI - Predicting gene essentiality using genome-scale in silico models. AB - Genome-scale metabolic models of organisms can be reconstructed using annotated genome sequence information, well-curated databases, and primary research literature. The metabolic reaction stoichiometry and other physicochemical factors are incorporated into the model, thus imposing constraints that represent restrictions on phenotypic behavior. Based on this premise, the theoretical capabilities of the metabolic network can be assessed by using a mathematical technique known as flux balance analysis (FBA). This modeling framework, also known as the constraint-based reconstruction and analysis approach, differs from other modeling strategies because it does not attempt to predict exact network behavior. Instead, this approach uses known constraints to separate the states that a system can achieve from those that it cannot. In recent years, this strategy has been employed to probe the metabolic capabilities of a number of organisms, to generate and test experimental hypotheses, and to predict accurately metabolic phenotypes and evolutionary outcomes. This chapter introduces the constraint-based modeling approach and focuses on its application to computationally predicting gene essentiality. PMID- 18392987 TI - Comparative approach to analysis of gene essentiality. PMID- 18392988 TI - [Continuing medical education in neurology: a necessary challenge]. AB - INTRODUCTION: The knowledge and skills acquired in the neurology specialty during post-graduate studies must be developed later as a specialist. Exponential growth of the scientific knowledge and its practical application make lifelong learning essential for optimal development of our professional activity. REVISION: The most relevant aspects of the learning and continued development of the neurologist have been reviewed. Current models and different proposals made by national and international medical organizations and societies on continuing medical education and the continuing professional development are presented and analyzed. CONCLUSION: Several aspects regarding the need for continuing medical education and the continuing professional development are discussed. Included among these are whether continuing education should be a voluntary commitment by each professional or an obligation regulated by specialist organizations. We have the opportunity to play a leading role in the changes that will affect us in this field. PMID- 18392989 TI - [Focal presentation of progressive multifocal leukoencephalopathy: first manifestation of human immunodeficiency virus infection]. AB - INTRODUCTION: Progressive multifocal leukoencephalopathy (PML) is a subacute demyelinating disorder of central nervous system caused by an opportunistic papovavirus infection of oligodendroglial cells, the JC virus. This virus remains latent in the human host. Their antibodies are detectable in 70% of healthy adult subjects. It is reactivated by different diseases and medical therapies impairing cellular immunity. CLINICAL CASE: A 35 year-old man, smoker, allergic to penicillin and derivatives, with a history of endovenous drug abuse, was admitted to Neurology Department due to a progressive left facial palsy and hand weakness. An alteration in the cellular immune ratio (CD4/CD8 0.07) and a positive human immunodeficiency virus serology were demonstrated. Highly active antiretroviral therapy (HAART) was started. A unique wide hyperintense subcortical right temporoparietal lobe lesion was shown by a cerebral magnetic resonance imaging (MRI), suggesting a differential diagnosis between PLM and low grade glial tumor. Cerebrospinal fluid study was normal and the polymerase chain reaction (PCR) was negative for JC virus. A steroataxic cerebral biopsy was made in order to establish a correct diagnosis due to patient's age and the good response to treatment. The pathology study showed changes suggestive of PLM and a positive PCR test for JC virus. DISCUSSION: We report this case to suggest that inva- sive diagnosis methods may be useful to differentiate between PLM and low grade glial tumors and to establish a correct prognosis. HAART is becoming a good treatment option because it seems to make long term survival longer, as is the case of this patient who is practically asymptomatic after more than 2 years of evolution. PMID- 18392990 TI - Validation of mechanically-assisted sodium dodecyl-sulphate elution as a technique to remove pellicle protein components from human enamel. AB - The salivary film, denoted the pellicle, formed on oral surfaces is of great importance for oral health and comfort. The present study describes mechanically assisted sodium dodecyl sulphate (SDS) elution of the in vivo pellicle formed on human enamel and visualisation of the desorbed pellicle proteins using two dimensional gel electrophoresis (2-DE). To verify this removal of the pellicle, a combined mechanical and surfactant procedure was additionally performed on an in vitro pellicle formed on human enamel, and the effectiveness was validated by mechanical removal in combination with HCl. As indicated by protein quantitation and one dimensional gel electrophoresis, rubbing with polyamide fibre pellets soaked in a 0.5% SDS solution was optimal for completely removing the adsorbed proteins from the enamel surface, and yet provided separation of the proteins by 2-DE to enable identification in future studies. PMID- 18392991 TI - Rheology of biofilms formed at the surface of NF membranes in a drinking water production unit. AB - In this study, the mechanical properties of biofilms formed at the surface of nano-filtration (NF) membranes from a drinking water plant were analysed. Confocal laser scanning microscopy observations revealed that the NF biofilms formed a dense and heterogeneous structure at the membrane surface, with a mean thickness of 32.5 +/- 17.7 mum. The biofilms were scraped from the membrane surface and analysed in rotation and oscillation experiments with a RheoStress 150 rotating disk rheometer. During rotation analyses, a viscosity decrease with speed of shearing characteristic of rheofluidification was observed (eta = 300 Pa s for y = 0.3 s(-1)). In the oscillation analyses with a sweeping of frequency (1 100 Hz), elasticity (G') ranged from 3000 to 3500 Pa and viscosity (G'') from 800 to 1200 Pa. Creep curves obtained with an application of a shear stress of 30 Pa were viscoelastic in nature. The G(0) and eta values were, respectively, 1.4 +/- 0.3 x 10(3) Pa and 3.3 +/- 0.65 x 10(6) Pa s. The relationship between the characteristics of NF biofilms and the flow conditions encountered during NF is discussed. PMID- 18392992 TI - Synopsis of the International Gap Junction Conference in Elsinore, Denmark August 5-9, 2007. AB - This synopsis covers the main results and conclusions from the platform presentations during the International Gap Junction Conference. More detailed information is provided in the mini reviews on controversial scientific issues, short reports of research results and conference abstracts published in this issue of Cell Communication and Adhesion. PMID- 18392993 TI - Gap junction-mimetic peptides do work, but in unexpected ways. AB - Gap junction mimetic peptides containing sequences of the extracellular loops of connexins inhibit the de-novo formation of gap junction channels but do not impair the function of existing cell-cell channels. Recently, a flurry of publications appeared showing that such "GAP" peptides attenuate ATP release and/or surrogate measures of it. Although no direct effect on putative connexin "hemichannels" has ever been shown, the peptide effect has been used as diagnostic tool for demonstrating the existence of such channels. However, testing of the peptides on genuine unapposed membrane channels formed by connexins failed to reveal any inhibitory action of the peptides on channel activity. Instead, membrane channels formed by the unrelated pannexin1 were inhibited in the same concentration range as described for the release of ATP. Consequently, rather than indicating connexin involvement in ATP release, the GAP peptide effects represent supporting evidence for a role of pannexin1 in this process. PMID- 18392995 TI - Proceedings of the International Gap Junction Conference. August 5-9, 2007. Elsinore, Denmark. PMID- 18392994 TI - Mimetic peptides as blockers of connexin channel-facilitated intercellular communication. AB - There is a dearth of chemical inhibitors of connexin-mediated intercellular communication. The advent of short "designer" connexin mimetic peptides has provided new tools to inhibit connexin channels quickly and reversibly. This perspective describes the development of mimetic peptides, especially Gap 26 and 27 that are the most popular and correspond to specific sequences in the extracellular loops of connexins 37, 40 and 43. Initially they were used to inhibit gap-junctional coupling in a wide range of mammalian cells and tissues. Currently, they are also being examined as therapeutic agents that accelerate wound healing and in the early treatment of spinal cord injury. The mimetic peptides bind to connexin hemichannels, influencing channel properties as shown by lowering of electrical conductivity and potently blocking the entry of small reporter dyes and the release of ATP by cells. A mechanism is proposed to help explain the dual action of connexin mimetic peptides on connexin hemichannels and gap-junctional coupling. PMID- 18392996 TI - The role that regional therapy plays in the current day management of cancer that is confined to the extremities. Introduction. PMID- 18392997 TI - History of regional chemotherapy for cancer of the extremities. AB - Patients with recurrent cutaneous or soft tissue malignancies of the extremity provide a unique opportunity to evaluate therapy targeted to the isolated limb. The most common clinical presentation of recurrent extremity malignancy occurs in patients with melanoma. The extremity is the site of primary melanoma in half of patients with the disease 1, and of those with a primary melanoma of Breslow depth >or=1.5 mm, 15% will develop a local or in-transit recurrence 2. Palliation of extremity disease is important in these patients, as median survival after diagnosis of in-transit or locally recurrent disease >2 years 3, 4. Radical approaches to eradication of extremity recurrence are rarely used, although in a highly selected group of patients undergoing amputation, 42% 5-year survival was reported 5. As greater recognition of the palliative nature of extremity therapy has evolved, an emphasis on preservation of limb function has supplanted cure as a more realistic therapeutic goal. While occasional cure can be observed, it would be misleading to propose this as the likely outcome of eradication of recurrent extremity melanoma. Isolated limb perfusion (ILP) was developed as an alternative to amputation in patients with recurrent cancer of the extremity. The concept was that vascular isolation of the limb would allow delivery of higher (and potentially more effective) doses of chemotherapy to the disease in the limb than could be achieved with systemic therapy. PMID- 18392998 TI - Effectiveness of regional chemotherapy with TNF-alpha/melphalan in advanced soft tissue sarcoma of the extremities. AB - Hyperthermic isolated limb perfusion with tumour necrosis factor alpha (TNF alpha) and melphalan was repeatedly reported to achieve extraordinarily high clinical remission rates in advanced and non-resectable soft tissue sarcoma of the limbs, thus avoiding imminent mutilation or amputation for most of those patients. With the limb being isolated throughout the extracorporal perfusion, high doses of recombinant TNF-alpha as well as melphalan can be applied. Basically, TNF-alpha directly affects the vasculature of the tumour and induces a severe inflammation with consecutive deterioration of the tumour capillaries. Furthermore, TNF-alpha increases the tumour-selective uptake of melphalan into the tumour cells thus leading to synergy of antivascular targeted treatment and antineoplastic effects of highest dose chemotherapy supplemented by hyperthermia. Meanwhile, a lot of sarcoma centres in Europe adopted this technique and established referral programmes for patients with non-resectable soft tissue sarcomas of the limbs. Despite these programmes many patients still do not get offered hyperthermic ILP with TNF-alpha and melphalan as a treatment option and modality. This article summarizes multimodality in treatment of soft tissue sarcoma of the limbs and reviews the current status of melphalan-based ILP with TNF-alpha (TM-ILP) and its results, to enable comparison and critical consideration of other treatment options. PMID- 18392999 TI - Current status of hyperthermic limb perfusion for in-transit melanoma. AB - In-transit disease is a unique form of regional lymphatic spread of melanoma that is considered an infrequent event although certain high-risk subgroups have been identified with higher incidence rates. Although this disease entity is associated with a high risk for distant relapse, regionally focused treatment of disease is important due to the high morbidity associated with in-transit disease. Isolated limb perfusion has been a utilized method of regional treatment since the 1950's. The technical aspects, indications, historical results, and toxicity of limb perfusion are reviewed. Finally, perfusion based treatment of in transit melanoma is an excellent model for studying novel agents and regimens in both the pre-clinical and patient care setting. PMID- 18393000 TI - Current status of isolated limb infusion with mild hyperthermia for melanoma. AB - PURPOSE: Recurrent disease confined to a limb is a frequently encountered clinical problem in patients with melanoma. Regional chemotherapy by isolated limb perfusion (ILP) provides effective treatment but is invasive, complex and costly. Isolated limb infusion (ILI) is a simple yet effective alternative to ILP. MATERIALS AND METHODS: ILI involves drug administration into a limb via percutaneously inserted catheters after vascular isolation of the limb has been achieved with a tourniquet. The infused drug is circulated for 30 minutes via a simple extracorporeal circuit incorporating a heater (to produce mild hyperthermia). RESULTS: Limb tumour remission rates are similar to those achieved by conventional ILP. ILI is well tolerated, and elderly patients and those with major medical co-morbidities and serious peripheral vascular problems can be treated. CONCLUSIONS: ILI with mild hyperthermia is an established alternative to hyperthermic ILP for patients with recurrent limb melanoma. It can also be used for patients with soft tissue sarcomas and a variety of serious, chronic dermatological conditions. PMID- 18393001 TI - The European approach to in-transit melanoma lesions. AB - The biological behavior of melanoma is unpredictable. Three to five per cent of melanoma patients will develop in-transit lesions and the median time to recurrence ranges between 13-16 months. At the time of recurrence the risk of occult nodal metastasis, with clinically negative regional lymph nodes, is as high as 50%. The risk of in-transit lesions depends on the tumor biology and not on the surgical approach to the regional lymph nodes. The high incidence of in transit lesions at the lower limb may be caused by the gravity and delayed lymphatic drainage. The treatment of limited disease is local excision, laser ablation, cryosurgery, while multiple in-transit lesions or bulky disease located in a limb can be successfully treated with regional chemotherapy, a therapeutic isolated limb perfusion or infusion with melphalan or a combination of melphalan and tumor necrosis factor (TNF) alpha. If local regional treatment or systemic dacarbazine based systemic treatment fails, novel systemic treatment strategies with vaccines, antibodies and gene therapy are currently investigated. PMID- 18393002 TI - Pharmacokinetics & drug resistance of melphalan in regional chemotherapy: ILP versus ILI. AB - Two forms of regional chemotherapy for the treatment of advanced melanoma or sarcoma of the extremity are isolated limb perfusion (ILP) and the more recently described isolated limb infusion (ILI). Melphalan is the most commonly employed agent in both ILP and ILI, although it is often used in conjunction with other cytotoxic and/or biologic therapies. While ILP and ILI are far more effective for the treatment of extremity disease than is systemic therapy, there is still significant room for improvement in outcomes, from the standpoint of both response rate and toxicity. An understanding of the pharmacokinetics of regional chemotherapy would allow for the prediction of tumor response and toxicity and therefore patient outcomes. In addition, elucidating the mechanisms of drug resistance would lead to opportunities to develop effective chemo-modulators that enhance the effectiveness of ILP and ILI. This paper reviews progress in these two key areas of active investigation. PMID- 18393003 TI - The role of hyperthermia in optimizing tumor response to regional therapy. AB - PURPOSE: To discuss the role of hyperthermia in optimizing the response to regional therapy for cancer. METHODS: A review of the current literature discussing mechanism of action, experimental models and prospective, randomized trials was performed. RESULTS: The mechanism of action of hyperthermia in combination with radiation and cytotoxic chemotherapy at the cellular and tissue levels is described. Data supporting the benefit of hyperthermia in conjunction with both regional, infusional chemotherapy, and locoregional radiation therapy is demonstrated. Several different histologic tumor types are covered, all with prospective evidence supporting the benefit of hyperthermia. CONCLUSIONS: Although delivery methods for hyperthermia can be complex and difficult to implement, the data support its benefit and further endeavors to include hyperthermia as a component of regional therapy should be encouraged. PMID- 18393004 TI - Electrochemotherapy: an emerging cancer treatment. AB - PURPOSE: The aim of this review article is to provide a concise overview of the pre-clinical development of electrochemotherapy (ECT), its present utility in clinical practice and to examine its potential application to therapeutic modalities in the future. RESULTS: Results from the ESOPE trial demonstrate an 85% objective response rate (ORR) in solid cutaneous and subcutaneous tumours of varying histologies, that would previously have been recalcitrant to conventional therapies. Experimentally, neoadjuvant immunogene therapy of primary cancers has been found to be effective against minimal residual disease in metastatic models. As such, combinations of electrogene delivery and electrochemothearpy offer exciting possibilities for both local and systemic control of heretofore incurable cancers. CONCLUSIONS: Electrochemotherapy is a quick, safe, inexpensive treatment modality that has been shown to give consistently reproducible results in the treatment of solid cutaneous and subcutaneous malignant tumours. To date, its clinical license has limited its application to a palliative setting. Future work includes looking to extend this therapeutic profile to the management of primary tumours and earlier stage disease, as well as examining the potential for combining electrochemotherapy with gene and immunotherapies and developing novel electrode designs to facilitate the application of this treatment to internal cancers. PMID- 18393005 TI - Toxicities associated with hyperthermic isolated limb perfusion and isolated limb infusion in the treatment of melanoma and sarcoma. AB - Hyperthermic isolated limb perfusion (HILP) and isolated limb infusion (ILI) may play a significant role in the treatment of patients with recurrent or in transit extremity melanoma or sarcoma that is unresectable. These procedures may be indicated when patients are otherwise faced with the possibility of a debilitating amputation. Not entirely benign treatment modalities, HILP and ILI can be associated with regional and systemic toxicities. We conducted a literature search of published studies using HILP and ILI for the treatment of extremity sarcomas and melanomas, and associated toxicities was performed. The regional toxicities of HILP and ILI are similar. The most common toxicities reported are mild to moderate. However, when severe regional toxicity occurs, albeit infrequently (<5%), fasciotomies or even amputation may be necessary. Some studies have showed a relationship between acute regional toxicities and long term regional morbidity. Systemic toxicity appears to be more frequent when TNF alpha is used in combination with other drugs during HILP, however the use of TNF alpha in the United States is limited to trials. Although regional toxicities are similar, systemic toxicity of ILI is minimal compared to HILP. ILI is easier to repeat, technically less complex, and may be more acceptable in infirmed patients. Long term morbidity and outcomes for ILI are still being evaluated. Both of these techniques may be suitable options in patients with unresectable advanced or recurrent, or in transit extremity melanoma or sarcoma. PMID- 18393006 TI - Laboratory models of regional chemotherapy. AB - Development of new treatment strategies and agents is a difficult and costly matter in oncology. Routinely drugs are tested in vitro on tumor cells which, however, may have limited predictive value for their activity in patients. Also, the pharmacokinetic behavior, intratumoral distribution, as well as toxic side effects, binding to other compounds and stability of an agent are very important in determining activity in the patient. More so, development and evaluation of surgical delivery methods, i.e. regional treatment strategies, cannot be tested in cell systems. Therefore animal models are crucial for the development of regional chemotherapy methodologies. To allow translation of the animal data to patients it is important that the animal model closely mimics the clinical setting as for instance is achieved with isolated limb perfusion. However, animal models remain limited in their use, as eventually the efficacy of the approach may be different in animals compared to patients. Here we describe the use of animal models for regional treatment of solid tumors. PMID- 18393007 TI - Future directions in regional treatment strategies for melanoma and sarcoma. AB - Hyperthermic isolated limb perfusion (HILP) with melphalan and more recently isolated limb infusion (ILI) with melphalan +/- dactinomycin are common treatment modalities for both in-transit melanoma of the extremity and advanced extremity sarcoma. In order to further optimize treatment, future research should focus on selection of appropriate patients, verification of a technique that produces consistent results while maintaining acceptable toxicity, and development of novel strategies and agents. Development of these novel agents and strategies has potential to not only improve the efficacy of regional chemotherapy but may also help guide future strategies for systemic treatment. PMID- 18393008 TI - Methods of fetal movement counting and the detection of fetal compromise. AB - Maternal perception of fetal movements is widely used as a marker of fetal viability and well-being. A reduction in fetal movements is associated with fetal hypoxia, increased incidence of stillbirth and fetal growth restriction (FGR). Therefore, a reduction in fetal movements has been proposed as a screening tool for FGR or fetal compromise. The problem of this approach is that there is no widely accepted definition of reduced fetal activity or 'alarm limits', and pregnant women are currently given a wide range of non-evidence-based advice. We have reviewed the background of published definitions and their potential usefulness in screening. A formal meta-analysis of these studies is not possible due to variation in methodology and definitions of reduced fetal movements. Assessment of fetal movements using formal fetal movement counting has shown equivocal results. Importantly, in all studies, there was a decrease in perinatal mortality suggesting a beneficial role for raising maternal awareness of fetal movements. Most studies implemented limits to define reduced fetal movements based on small groups of high risk pregnancies and obsolete counting methodology. A single case-control study developed 'normal limits' in a low risk population, and successfully implemented it prospectively for screening. At present, there is no evidence that any absolute definition of reduced fetal movements is of greater value than maternal subjective perception of reduced fetal movements in the detection of intrauterine fetal death or fetal compromise. Further investigation is required to determine an effective method of identifying patients with reduced fetal movements and to determine the best subsequent management. PMID- 18393009 TI - Genetic influences on smoking cessation and relapse in pregnant women. AB - Cigarette smoking during pregnancy continues to be a significant public health concern. Maternal smoking during pregnancy has been associated with low birth weight (<2500 g), fetal growth restriction, placental problems, pre-term delivery and spontaneous abortion. Mothers who smoke during pregnancy are twice as likely to give birth to low birth weight infants, and smoking during pregnancy is estimated to be responsible for 20-30% of all low birth weight infants. Smoking during pregnancy not only affects placental function, thus causing obstetrical complications, but nicotine also crosses the placenta and acts as a neuroteratogen. This in turn, elevates the risk of cognitive and auditory processing deficits, and has also been found to be negatively associated with long-term consequences on offspring behaviour. In addition, smoking has negative long-term health consequences for both mother and child, including respiratory conditions, cancer and cardiovascular problems. This review provides insight into the genetic influences on smoking behaviour in pregnant women. In particular, the roles of genes in the neurotransmitter pathways are highlighted. It also emphasises the need for further research in this area, and provides rationale for the importance of focusing on pregnant women who are highly motivated to quit when researching smoking behaviours in women. PMID- 18393010 TI - A review of the management of diseases of the Bartholin's gland. AB - Bartholin's abscess and cyst account for 2% of all gynaecological visits per annum. This article reviews the anatomy, epidemiology and pathophysiology of the Bartholin's gland. The management of Bartholin's abscess including antibiotic use in the early stage of the condition together with options for surgical intervention are discussed and in so doing the variation of clinical practice in other countries is considered. The use of the word catheter, carbon dioxide laser are compared with traditional surgical techniques of incision and drainage and marsupialisation. The outcomes of treatment particularly the high recurrence rate (2-25%) and the associated morbidity are reviewed. Finally the issue of cancerous pathology as an aetiology is highlighted in the postmenopausal age group. PMID- 18393011 TI - The treatment of urinary incontinence with Duloxetine. AB - Duloxetine is a Serotonin and Noradrenaline reuptake inhibitor, and is the first drug licensed for medical treatment of urodynamic stress incontinence. In this paper, we will briefly review the pharmacology, current evidence and controversies in relation to the use of duloxetine. PMID- 18393012 TI - Newborn brachial plexus injuries: The twisting and extension of the fetal head as contributing causes. AB - The exact mechanism of the causation of brachial plexus injury (BPI) has long been a matter of controversy. It is our opinion that the twisting and the extension of the fetal head, during the labour and delivery process, will increase the stretching of the neck, thus contributing to the labour forces as the cause of BPI. Our opinions are offered to other researchers and readers for their consideration of how the labour forces can cause BPI. PMID- 18393013 TI - Avoiding over diagnosis of shoulder dystocia. AB - In this study, we aimed to distinguish true shoulder dystocia from mere difficulty with delivery of the shoulder, by investigating the risk factors that lead to shoulder dystocia. Shoulder dystocia is a bony problem which occurs when either the anterior or, less commonly, the posterior fetal shoulder impacts on the maternal symphysis pubis or sacral promontory. Failure to apply the Royal College of Obstetricians and Gynaecologists (RCOG) clinical diagnostic criteria for shoulder dystocia has made it susceptible to over-diagnosis due to inclusion of all difficult shoulder deliveries, including those related to an inappropriate maternal position. This was a retrospective analysis of risk factors associated with 56 cases of shoulder dystocia which occurred in West Middlesex University Hospital between 2003 and 2004. The cases were analysed in two categories, good outcome and poor outcome, and compared with each other. The poor outcome had represented true shoulder dystocia. The incidence of shoulder dystocia increased from 0.94% in 2003 to 1.37% in 2004. However, the incidence of those with a poor outcome decreased from 45.4% of the whole shoulder dystocia group in 2003, to 17.6% in 2004 (p = 0.03). There were no clear diagnostic criteria documented in the notes for the condition other than the birth attendants' opinion and the turtle sign. This may either reflect over-diagnosis from increased awareness or possibly improvement in the outcome due to training and education. Interestingly, at least four risk factors were identified in each of the cases with poor outcome. A registrar conducting the delivery, forceps delivery for delayed second stage and the turtle sign were significantly common findings among the true shoulder dystocia group. Multiple risk factors can be a good predictor for the occurrence of shoulder dystocia. Applying the RCOG diagnostic criteria for shoulder dystocia may lead to improvement in diagnosis and therefore a better understanding of the risk factors and future management of shoulder dystocia. PMID- 18393014 TI - The views of obstetricians in the south-west of England on the use of prostaglandins and syntocinon in VBAC. AB - The aim of our study was to explore the views of consultant obstetricians in the south-west of England on the use of prostaglandin and oxytocin in women undergoing vaginal birth after caesarean delivery (VBAC), and assess how the perceived risk of uterine rupture in such a cohort of women affected their practice. A postal questionnaire was sent to randomly selected consultant obstetricians in the south-west of England. The response rate from the survey was 59.2%. The study showed that the willingness of consultants to use prostaglandin in VBAC was related to their length of practice. Some 89.7% of all respondents claim they would use syntocinon for augmentation in VBAC; 75% of all the responding consultants who have been in the post for longer than 10 years claim they would not use either of the agents in VBAC. We conclude that, despite concerns about the risks of increased uterine rupture in both induction and augmentation in labour of women undergoing VBAC, our study did not seem to support the fact that obstetricians' practices are influenced by this perceived risk. PMID- 18393015 TI - Ligation of internal iliac artery for severe obstetric and pelvic haemorrhage: 10 year experience with 11 cases in a university hospital. AB - Bilateral iliac artery ligation is an effective second-line procedure to control massive obstetric and gynaecological haemorrhage, it is life-saving in certain cases and has the advantage that it preserves fertility, which is particularly important in young women of low parity. Bilateral ligation of internal iliac arteries is a life-saving procedure in cases of massive obstetric haemorrhage. It has the advantage of preserving fertility compared with the commonly performed emergency obstetric hysterectomy. It has also proved effective in cases of secondary pelvic haemorrhage in gynaecological operations. We report here a case series of 11 women who had bilateral internal iliac artery ligation for severe pelvic haemorrhage in a period of 10 years in our department. Ten of them were obstetric cases and one was a case of haemorrhage following vaginal hysterectomy. The outcome was favourable in 10 out of the 11 cases and there was only one case of massive postpartum haemorrhage where the procedure failed and required an emergency hysterectomy. PMID- 18393016 TI - Obstetric management and perinatal outcome of extreme prematurity: a retrospective study. AB - The aim of this retrospective study is to evaluate the obstetric management and perinatal outcome of extreme prematurity (22-27 weeks) in a busy teaching hospital between April 2004 and March 2005. A total of 57 babies were delivered in 49 women representing 0.9% of total births. A total of 67% of babies were transferred to the NICU and the survival rate for those who were admitted to the NICU was 47%. The overall survival rate for all births at hospital discharge was 32%. Caesarean section was carried out in 32% of the 49 mothers. The overall survival at discharge in these babies was 50%. Only 12.5% of babies delivered by caesarean section at less than 27 weeks survived as compared with 70% survival rate at 27 weeks. There was no survival among babies delivered by caesarean section below 26 weeks. Gestational age of the neonate was the single most important parameter related to survival at 7 days, 30 days and 90 days/discharge, even after adjusting for other parameters like birth weight, sex of baby and maternal chorioamnionitis. Birth weight was an independent risk factor for survival, with a birth weight of 900 g; the survival rate was significantly higher. Hypertensive disorder was the most common maternal medical complication and responsible for 7/16 caesarean sections. The study highlighted the importance of the multidisciplinary team management and the involvement of parents in the decision regarding management of these very pre-term babies, during labour and after delivery. PMID- 18393017 TI - Amniocentesis counselling: a role for the midwife-practitioner. AB - Recent publication of an evidence-based clinical guideline by the Royal College of Obstetricians and Gynaecologists (RCOG 2005) for invasive testing in pregnancy stimulated a review of our prenatal diagnosis counselling service. This coincided with a reduction in the hours worked by obstetric trainees and a need to streamline antenatal care. We arranged for a senior midwife (KE) with extensive experience in general midwifery and fetal medicine to undergo additional training in counselling for amniocentesis. She then took over the running of the counselling service supported by an in-house care pathway. She had open access to a consultant (RF) for advice. A review of the case notes of 60 consecutive women who attended for counselling showed that the midwife followed the guidelines extremely closely both in terms of process and quality of the documentation. A total of 58 of the women were counselled solely by the midwife-practitioner. Only two required additional counselling by the consultant. Of a subset of 27 women surveyed by telephone questionnaire, only two (7.4%) were surprised to have been counselled by a midwife; 25 (93%) said the counselling was excellent or good; 17 (63%) said they would prefer to see a midwife in any future pregnancy and only one woman said she would prefer to see a doctor. PMID- 18393018 TI - The safety of elective caesarean section for the prevention of mother-to-child transmission of HIV-1. AB - The benefits of elective caesarean section (ELCS) for the prevention of mother-to child transmission (PMTCT) of HIV are now well known. However, the benefits of such a procedure must be weighed against the risks involved. This study examines the risks of ELCS for PMTCT at the University of Maiduguri Teaching Hospital, Maiduguri, Nigeria, from January-December 2006 inclusive. A total of 52 HIV positive mothers who opted for ELCS for PMTCT were prospectively followed-up for any intra- and postoperative complication. A total of 52 cases of ELCS of HIV negative mothers during the same period of study were used as controls. The HIV positive mothers were found to be younger and of lower parity than the uninfected women. Unlike the uninfected mothers, they did not have co-morbid conditions, such as diabetes mellitus and severe pregnancy induced hypertension. There was no significant difference in intra- and postoperative morbidities between the two groups of mothers. The fetal outcome in terms of Apgar scores and birth weight was significantly better among the HIV-infected women. It is recommended that HIV infected mothers in this environment should be offered the option of ELCS for the purpose of prevention of HIV from mother to child. PMID- 18393019 TI - Outcome of emergency caesarean sections in Calabar, Nigeria: Impact of the seniority of the medical team. AB - We set out to assess the maternal outcome of emergency caesarean sections in University of Calabar Teaching Hospital (UCTH) in relationship to the seniority and experience of medical personnel involved in the operation. This was a review of 349 cases of emergency caesarean sections in UCTH over a 2-year period (January 2000-December 2001). The sociodemographic data, antenatal booking status and clinical condition of the patients as well as the seniority of the medical staff who participated in the operations were extracted from the case notes of the patients. A total of 280 (80.0%) booked patients and 69 (20.0%) unbooked mothers were delivered by emergency caesarean sections. The consultants performed only 16.4% of the emergency caesarean sections, while the residents performed 83.6%. Similarly consultant anaesthetists administered anaesthesia in 12.9% of the cases. A total of 12 maternal deaths occurred from emergency caesarean sections in which six (1.7%) were due to sepsis, three (0.8%) due to severe haemorrhage and three (0.8%) due to anaesthetic complications. None of the deaths occurred in operations in which consultant obstetricians and anaesthetists were involved. The intraoperative blood loss was more in operations performed by the residents than in those performed by the consultants. Similarly, morbidity was higher in patients operated upon by residents. The maternal morbidity and mortality associated with emergency caesarean sections in UCTH would be reduced significantly if consultant obstetricians and anaesthetists were more involved in these operative procedures. PMID- 18393020 TI - Antepartum anxiety and depressive symptoms: A study of Nigerian women during the three trimesters of pregnancy. PMID- 18393021 TI - Epidemiological factors influencing IVF outcome: Evidence from the Yale IVF program. AB - Age, BMI, lifestyle, menstrual status and obstetric history can modulate the endocrine system and, therefore, have been hypothesised to play a role in in vitro fertilisation (IVF) outcome. We designed a retrospective study, set in a medical school hospital. We evaluated the medical files of 297 infertile women who underwent laparoscopy and consecutive IVF-ET treatment in the Yale IVF unit between 1996 and 2002. The study group consisted of 151 women who conceived after IVF-ET and the control group of 146 women who underwent 288 IVF-ET cycles without pregnancy. The main outcome measure was the impact of epidemiological factors on the IVF outcome. There was no association between IVF outcome and race, BMI, age at menarche, length of cycle, duration and amount of flow, menstrual symptoms, other medical problems, medical history of allergies, and family history of endometriosis and cancer. We found that the degree of smoking and alcohol use was not a factor when comparing women with and without pregnancy after IVF (34.5% vs 29.5%, and 33.7% vs 27%, respectively). The rate of duration of infertility tended to be lower in pregnant women (35.9+/-23.4 months) vs (42.3+/-30.2) non pregnant women. As expected, we also confirmed the inverse association between the age of women and IVF outcome. Overall, body attributes, lifestyle, family history, menstrual and reproductive factors were not related to IVF-ET outcome. PMID- 18393022 TI - Management of suspected ectopic pregnancy: An audit from East Kent NHS Hospital Trust. AB - This is a retrospective case note analysis of 105 suspected cases of ectopic pregnancies that underwent laparoscopy within a stipulated 12-month period, September 2001 to August 2002. The management and care of these women was reviewed using the Royal College of Obstetricians and Gynaecologists' (RCOG) guidelines on management of ectopic pregnancies as well as local guidelines developed from the RCOG guidance. Other criteria used to review patient care were derived from the National Confidential Enquiry into Patient Outcomes and Death (NCEPOD). A large number of procedures (51.13%) which include laparoscopy, laparotomy and combined procedure were performed by the consultant. Approximately 90.19% of all procedures were performed during official working hours, which is in keeping with the National Confidential Enquiry into Patient Outcome and Death (NCEPOD) recommendation. Other standards and criteria used were the use of anti-D immunoglobulin and the use of thromboprophylaxis. A total of 86.7% of non sensitised Rhesus negative women received anti-D immunoglobulin; 96% of suspected ectopic pregnancies were diagnosed within 96 h of initial presentation. In 80% of cases, betahCG results were available within 24 h of request. Only 26% of ectopic pregnancies were successfully treated using laparoscopy, indicating that even though operative laparoscopy is the preferred approach, operative complications and haemoperitoneum, can lead to changes in surgical approach. The results of the audit identified areas for improvement and offers a practical solution through recommendations based on the audit process. PMID- 18393023 TI - An experimental study on the effect of different types of textiles on conception. AB - To study the effect of different types of textiles on conception. A total of 35 female dogs were divided into five equal groups: four test and one control. Each of the four test groups was dressed in one type of textile underpants made of either 100% polyester, 50/50% polyester-cotton mix, 100% cotton, or 100% wool. The pants were worn for 12 months. The dogs were followed during this period and for 6 months after removal of the pants by measuring the serum oestradiol 17beta and progesterone and by mating. Electrostatic potentials were also measured on the textile-covered skin. Eight bitches wearing polyester-containing textile showed diminished serum progesterone in the oestrus of the oestrous cycle, and did not conceive on mating or insemination. Five months after pants had been removed, serum progesterone of the eight dogs had normalised, and they conceived. Electrostatic potentials were detected on the skin of all dogs dressed in polyester-containing textile. The low serum progesterone and non-conception of the eight dogs could point to anovulation and failure of luteinisation. It is suggested that the electrostatic potentials detected on the skin create an 'electrostatic field' that inhibits the ovarian function. However, the effect proved to be reversible. PMID- 18393025 TI - A negative D-dimer does not exclude venous thromboembolism (VTE) in pregnancy. PMID- 18393024 TI - Patterns of menopausal symptoms and adaptive ability in a rural population in South-east Nigeria. AB - We set out to assess menopausal symptoms and the adaptability to them in a Nigerian population. This was a cross-sectional survey using a self-administered questionnaire and supplemented with focus group discussion carried out on a rural postmenopausal population in South-east Nigeria. Data were analysed using the epi Info version 3.3.2. Data from 186 were analysed. The mean age of women at the time of interview was 49.6+/-6.3 years. Mean and median ages at menopause were 45.47+/-5.5 years and 47 years, respectively. The major symptoms at menopause were: hot flushes 36 (58.1%), sweating 25 (40.3%), urinary frequency 24 (38.7%), vaginal dryness, discomfort or discharge 22 (35.5%), lack of concentration 17 (27.4%) and irritability 15 (24.2%). Widowhood and the empty nest syndrome but not educational attainment imparted negatively on the ability of the women to cope with the menopausal symptoms. Menopausal symptoms are a health concern to the Christian population in South-east Nigeria. Doctors working in this environment should utilise the opportunity of clinical consultation to raise and discuss these symptoms with postmenopausal women. PMID- 18393026 TI - Early fundoscopy, magnetic resonance imaging and venometry in the diagnosis of venous sinus thrombosis. PMID- 18393027 TI - Pregnancy outcome with Marfan's syndrome and aortic root replacement. PMID- 18393028 TI - Pregnancy and delivery in a patient with hypoplastic lung and dyspnoea. PMID- 18393029 TI - Unusual cause of itching in a pregnancy (cholangiocarcinoma). PMID- 18393030 TI - Successful pregnancy in Fredrickson type I hyperlipidaemia. PMID- 18393031 TI - Successful outcome of a pregnancy complicated by type 3 cryoglobulinaemia with negative Hepatitis C virus (HCV) serology. PMID- 18393032 TI - Postpartum diagnosis of renal carcinoma in pregnancy: Important learning points. PMID- 18393033 TI - Improving the outcome of pregnancy in essential thrombocythaemia: The role of aspirin. PMID- 18393034 TI - Successful pregnancy outcome after pre-term premature rupture of membranes at 17 weeks' gestation. PMID- 18393035 TI - Placenta percreta. PMID- 18393036 TI - Postoperative obstruction following caesarean section possibly due to non-closure of the peritoneum. PMID- 18393037 TI - Schwannoma of the bladder: A rare pelvic tumour. PMID- 18393038 TI - Vulval mass as the first manifestation of non-Hodgkin's lymphoma. PMID- 18393039 TI - Massive intra-abdominal haemorrhage due to spontaneous bleeding from fibroids in a post-menopausal woman. PMID- 18393040 TI - Outpatient management of incarcerated ring pessary: Use of orthopaedic bone cutters. PMID- 18393041 TI - Can uterine artery embolisation affect endometrial perfusion and function. PMID- 18393042 TI - Ovarian hyperthecosis and response to antiandrogens: An uncommon presentation of a common disorder. PMID- 18393043 TI - Massive ovarian cyst haemorrhage with haemoperitoneum as a complication of long term anticoagulation. PMID- 18393044 TI - Incarcerated appendices epiploicae through the posterior vaginal defect secondary to a ring pessary. PMID- 18393045 TI - Persistent gestational trophoblastic neoplasia following a term vaginal delivery. PMID- 18393046 TI - Brow presentation does not mean caesarean section. PMID- 18393050 TI - Presence and persistence of psychotic symptoms in cocaine- versus methamphetamine dependent participants. AB - The primary objective of this study was to compare and contrast psychotic symptoms reported by cocaine- and methamphetamine-dependent individuals. Participants included 27 cocaine-dependent and 25 methamphetamine-dependent males, as well as 15 cocaine-dependent and 18 methamphetamine-dependent females. After screening, participants were excluded if they met criteria for any Axis I diagnosis other than nicotine dependence, or methamphetamine or cocaine dependence (ie, participants had to use either methamphetamine or cocaine but were excluded if they met dependence criteria for both). The participants were administered the newly developed Psychotic Symptom Assessment Scale (PSAS), which assesses psychotic symptoms. A high proportion of both cocaine- and methamphetamine-dependent men and women reported delusions of paranoia and auditory hallucinations. However, during the abstinent and intoxicated conditions, methamphetamine-dependent men and women were more likely than cocaine dependent men and women to report psychotic symptoms. Future studies will compare psychotic symptoms reported by non-dependent recreational stimulant users to stimulant-dependent individuals. PMID- 18393051 TI - Identifying methamphetamine users at risk for major depressive disorder: findings from the methamphetamine treatment project at three-year follow-up. AB - Little is known about risk factors for depression in methamphetamine users. Using data from 526 adults in the largest psychosocial clinical trial of methamphetamine users conducted to date, this study examined clinical, demographic, and substance use characteristics that predict the presence of a diagnosis of major depressive disorder (MDD) three years after treatment for methamphetamine dependence. The results indicate that two risk factors predict a diagnosis of MDD: a Beck Depression Inventory total score greater than 20, and one or more prior suicide attempts. These risk factors identify methamphetamine users who may benefit from early interventions for psychiatric symptoms. PMID- 18393052 TI - Predictors of cardiovascular response to methamphetamine administration in methamphetamine-dependent individuals. AB - The goal of the present investigation was to determine predictors of cardiovascular response to methamphetamine administrated in the laboratory. Heart rate (HR) and blood pressure (BP) were measured at baseline and at several time points following the administration of methamphetamine or saline placebo. One-way ANOVA was used to determine the differences between female and male subjects in their cardiovascular response. In male subjects, linear regression and one-way ANOVA were used to determine the influence of potential predictors on cardiovascular response, including age, weight, drug use indicators, concurrent use of other substances, route of administration, and race. Methamphetamine administration provoked significant increases in HR and BP, as compared to placebo. Female gender was associated with larger peak change in diastolic BP following administration. Baseline HR and BP were found to be strong predictors of cardiovascular response to methamphetamine administration in male subjects. Lifetime use and recent use of methamphetamine and nicotine did not predict cardiovascular response to methamphetamine. Recent alcohol use was associated with increased peak change in diastolic BP. Also, current use of cannabis was negatively correlated with peak HR change. Male cannabis users show lower peak change in HR as compared to non-cannabis users. As compared to methamphetamine smokers, intravenous users demonstrated higher peak change in diastolic BP following drug administration. Race did not have a significant effect on cardiovascular response. Taken together, these findings may help in the prevention and treatment of cardiovascular events in a population at high risk of premature morbidity and mortality. PMID- 18393053 TI - The color of meth: is it related to adverse health outcomes? An exploratory study in Tijuana, Mexico. AB - In a study of injection drug users (IDUs) in Tijuana, Mexico, logistic regression identified factors associated with injection of colored vs. clear methamphetamine in the prior six months (N = 613). Colors injected most often were clear (50%), white (47%), yellow (2%), and pink (1%). IDUs injecting colored meth were more likely to experience recent abscesses (34%) compared to those injecting clear meth (24%; p = 0.008), an association that persisted after adjusting for confounders. Market characteristics, possibly relating to purity or adulterants, may be associated with abscesses among methamphetamine injectors. Further study is needed to confirm and determine the mechanism of this association to better inform prevention messages. PMID- 18393054 TI - Long-term treatment with buprenorphine/naloxone in primary care: results at 2-5 years. AB - To examine long-term outcomes with primary care office-based buprenorphine/naloxone treatment, we followed 53 opioid-dependent patients who had already demonstrated six months of documented clinical stability for 2-5 years. Primary outcomes were retention, illicit drug use, dose, satisfaction, serum transaminases, and adverse events. Thirty-eight percent of enrolled subjects were retained for two years. Ninety-one percent of urine samples had no evidence of opioid use, and patient satisfaction was high. Serum transaminases remained stable from baseline. No serious adverse events related to treatment occurred. We conclude that select opioid-dependent patients exhibit moderate levels of retention in primary care office-based treatment. PMID- 18393055 TI - Physical pain and associated clinical characteristics in treatment-seeking patients in four substance use disorder treatment modalities. AB - Physical pain among persons seeking treatment for substance use disorders (SUD) and characteristics associated with pain were examined in a secondary analysis of data from the Drug Abuse Treatment Outcome Study (DATOS), a multi-site treatment outcome study. Patients (N = 7,876) in four treatment modalities - long-term residential (LTR), short-term inpatient (STI), outpatient methadone treatment (OMT), and outpatient drug-free (ODF) - reported severity of physical pain experienced during the preceding 12 months. Moderate to severe physical pain was reported by 21.2% of LTR patients, 26.8% of STI patients, 33.6% of OMT patients, and 17.6% of ODF patients. Individuals with and without physical pain were compared across treatment modalities. Patients with pain were more likely to report weekly heroin use [aOR = 1.73 (1.44-2.08)], weekly narcotics use [aOR = 1.43 (1.18-1.74)] and greater depressive symptoms [aOR = 1.30 (1.21-1.38)]. These findings support the presence of a sizable proportion of SUD patients with pain who may benefit from pain assessment as part of their SUD treatment. PMID- 18393057 TI - Contextual influences on polydrug use among young, low-income women: effects of neighborhood and personal networks. AB - This study assessed contextual risks for polydrug use in a triethnic sample (non Hispanic white, African American, Hispanic) of young women with a low income. For the current analysis, a total of 712 young women aged 18 to 31 years who sought care in state-funded family planning clinics in southeast Texas from December 2001 to May 2003 participated in the survey. The main outcome of the study was the number of illicit drugs (including marijuana, MDMA [ecstasy], crack cocaine, and other hard drugs) used in the last 12 months. Of the 712 subjects, 198 (28%) reported using illicit drugs in the past 12 months. Neighborhood socioeconomic status was significantly associated with drug use in a bivariate model. The proportion of women living in the most advantaged neighborhoods who reported drug use was more than twice that of women living in the most disadvantaged neighborhoods. However, the significance of neighborhood socioeconomic status was eliminated after controlling for ethnicity or for personal network characteristics in a multivariate ordinal logistic regression model. In contrast, in multivariate models, personal network indicators, such as a larger number of monthly contacts with friends (odds ratio [OR] = 1.32, 95% confidence interval [CI] = 1.11, 1.56) and a larger number of friends who used illicit drugs (OR = 1.47, 95% CI = 1.33, 1.62) were associated with increased drug use. In addition, not being currently married (vs. being married) (OR = 2.73, 95% CI = 1.44, 5.16) was associated with a larger number of drugs used in the last 12 months. In conclusion, we found that neighborhood socioeconomic status was not directly associated with more drug use when controlling for ethnicity or for personal network characteristics. Personal networks may mediate the relationships between neighborhood and drug use. Strategies to reduce polydrug use should target personal networks where friends use illicit drugs. PMID- 18393056 TI - Gender differences in mental health characteristics and gambling among African American adolescent gamblers. AB - This study explores gender differences in lifetime and recent substance use/internalizing behavior, childhood externalizing behavior, and gambling preferences among African-American youth gamblers. Data are from a prospective study of a community sample of 452 urban African-American youths who began at entry into first grade and were followed for ten years. Gambling was associated with high teacher ratings of childhood externalizing behaviors among males and with high parent ratings of childhood impulsivity and hyperactivity among both genders. Internalizing behavior was associated with female gambling. No male female differences in substance use/lifetime conduct disorder among gamblers were noted. Gambling preferences/frequency differed across genders. PMID- 18393058 TI - Research- and community-based clinicians' attitudes on treatment manuals. AB - We assessed the attitudes of 18 research- and 22 community-based substance abuse clinicians on treatment manuals. Research and community clinicians exhibited favorable attitudes toward manuals, and the majority (72% and 77%, respectively) reported an interest in learning more about substance use disorder (SUD) treatment manuals. Among community clinicians, greater years of experience was significantly associated with less favorable attitudes toward treatment manuals. Research clinicians endorsed significantly higher ratings for the importance attached to "theoretical rationale/overview" and "main session points to address" than community clinicians. Findings suggest that community SUD clinicians are already familiar with and have positive attitudes toward manuals, but specific subgroups have concerns that should be addressed. PMID- 18393059 TI - Impact of a brief training on medical resident screening for alcohol misuse and illicit drug use. AB - Educational initiatives are needed to improve primary care substance use screening. This study assesses the impact on 24 medical residents of a 2.5-day curriculum combining experiential and manual-based training on screening for alcohol misuse and illicit drug use. A retrospective chart review of new primary care outpatients demonstrated that nearly all were asked about current alcohol use before and after curriculum participation. Adherence to national screening guidelines on quantification of alcohol consumption modestly improved (p < .05), as did inquiry about current illicit drug use (p < .05). Continued efforts are needed to enhance educational initiatives for primary care physicians. PMID- 18393060 TI - The association between earlier marijuana use and subsequent academic achievement and health problems: a longitudinal study. AB - In this prospective longitudinal study, the authors investigated the association between marijuana use over a period of 13 years and subsequent health problems at age 27. A community sample of 749 participants from upstate New York was interviewed at mean ages of 14, 16, 22, and 27 years. Marijuana use over time was significantly associated with increased health problems by the late twenties, including respiratory problems, general malaise, neurocognitive problems, and lower academic achievement and functioning. Effective prevention and intervention programs should consider the wide range of adverse physiological and psychosocial outcomes associated with marijuana use over time. PMID- 18393061 TI - Use of dronabinol for cannabis dependence: two case reports and review. AB - Marijuana is the most commonly used illicit drug in the United States and throughout the world. Despite this, the number of laboratory studies that have assessed pharmacologic agents to target cannabis withdrawal symptoms or reduce the reinforcing effects of marijuana has been modest. Unlike alcohol, cocaine, opiates, or nicotine, there has been a minimal number of clinical pharmacologic treatment trials that have targeted marijuana use. Based on recent laboratory studies, dronabinol (delta-9-tetrahydrocannabinol) has been shown to reduce cannabis withdrawal symptoms and the subjective effects of marijuana. Given that agonist agents have been found to be effective for opiate and nicotine dependence, the clinical utility of dronabinol for cannabis dependence is a reasonable approach. Two case reports using dronabinol are presented. The potential benefit, as well as questions that arise from the use of this medication in cannabis-abusing populations, is presented. Also, future areas of research that might be explored are discussed. PMID- 18393062 TI - A pilot open-label trial of ropinirole for cocaine dependence. PMID- 18393063 TI - Polychlorinated biphenyls in stormwater runoff entering the tidal Anacostia River, Washington, DC, through small urban catchments and combined sewer outfalls. AB - To investigate the loadings, solid-water partitioning, transport dynamics, and sources of polychlorinated biphenyls (PCBs) in urban stormwater runoff entering into the lower tidal Anacostia River, which flows south of Washington, DC, USA, storm and base flow samples were collected in six branches. Stormwater runoff contained elevated levels of PCBs (9.82 to 211 ng/L) higher than base flow by up to 80-fold. The present study suggests that input of PCBs from Lower Beaverdam Creek is likely to be greater than those from the two major branches (Northeast and Northwest Branches) that were believed as primary source areas. PCBs in storm flow were significantly enriched in the particle phase, which accounted for more than 90% of the total PCBs. Particles were the primary vector transporting PCBs into the Anacostia River, suggesting that removal of particles in stormwater runoff using best management practices (BMPs) such as post treatment system likely decrease PCBs significantly. PCB congener patterns found in stormwater samples clearly explain stormwater runoff is a major transport pathway adding substantial amount of PCBs to the tidal Anacostia River. PMID- 18393064 TI - The effects of pyrene exposure on exercise performance, muscle contraction, and mitochondrial O(2) consumption in the leopard frog (Rana pipiens). AB - The present study exposed frogs to the PAH pyrene and measured exercise performance, muscle contractile ability, mitochondrial O(2) consumption, and membrane potential. Leopard frogs, Rana pipiens, were exposed for seven days in control or pyrene saturated water aquaria. Frogs were randomly placed into one of four groups: (i) pyrene-exposed exercised, (ii) control, exercised, (iii) pyrene exposed, non-exercised, and (iv) control, non-exercised. Following the acute exposure, exercise duration, muscle contractile ability, blood gases and pH, glycogen levels, crossbridge formation and contractile length, and mitochondrial respiration were measured. The data revealed that pyrene exposure produced many adverse effects in Leopard frogs including significant reductions in exercise performance, muscle contractile ability, and alterations to muscle mitochondrial O(2) consumption and membrane potential. These data suggest that PAH exposure may limit survival for frogs in the wild by limiting foraging, mating, and predator avoidance behaviors. PMID- 18393065 TI - Assessment of diesel contamination in groundwater using electromagnetic induction geophysical techniques. AB - Determining hydrocarbon plumes in groundwater is typically accomplished through the installation of extensive monitoring wells. Issues of scale and site heterogeneities tend to introduce errors in delineating the extent of contamination and environmental impact. In this study, electromagnetic induction survey was investigated as an alternative technique for mapping petroleum contaminants in the subsurface. The surveys were conducted at a coal mining site near Gillette, Wyoming, using the EM34-XL ground conductivity meter. Data from this survey were validated with known concentrations of diesel compounds detected in groundwater from the study site. Groundwater data correlated well with the electromagnetic survey data, which was used to generate a site model to identify subsurface diesel plumes. To our knowledge, this is one of the first studies to use electromagnetic survey techniques for mapping hydrocarbon contamination in groundwater. Results from this study indicate that this geophysical technique can be an effective tool for assessing subsurface petroleum hydrocarbon sources and plumes at contaminated sites. PMID- 18393066 TI - Producing biodiesel using whole-cell biocatalysts in separate hydrolysis and methanolysis reactions. AB - This research examined the effect of using separate hydrolysis and methanolysis reactions for biodiesel production using a whole-cell biocatalysts derived from Rhizopus Oryzae (ATCC 10260). Biodiesel yield from separate hydrolysis and methanolysis was compared to transesterification reactions where both hydrolysis and methanolysis reactions occur in the same reactor. All reactions were conducted at room temperature. The effect of substituting ethanol for methanol was also studied. Separating the hydrolysis and methanolysis reactions did not significantly increase biodiesel yields; however, this approach successfully converted about 99% of triglycerides into fatty acid methyl esters (FAME) and free fatty acids (FFA). Use of ethanol in place of methanol did not significantly improve the biodiesel yield. However, there is evidence that ethanol may either esterify FFA more quickly than methanol, or result in a more stable ester. The best biodiesel yield was about 90% when a transesterification reaction using methanol was followed by one hydrolysis and one ethanolysis reaction; however, this is only slightly higher than the 88% biodiesel yield of two transesterification reactions in series (using methanol as alcohol). PMID- 18393067 TI - Study on a three-dimensional testing method coupling with a leaching behavior model for solidified waste matrix. AB - The objective of this study was to develop a three-dimensional leaching method to understand the diffusion behavior of a solidified waste matrix. A cylindrical solidified waste matrix with isotope lead compounds used as a tracer was used to demonstrate the diffusion phenomenon. The leaching test method was coupled with the mathematical diffusion model derived from Duhamel's theorem to control the time-dependent conditions and compute the mass diffusivity and mass generation rate constant of the target pollutants and also simulate the pollutants leached from solidified waste matrix. The simulation value is in fair agreement with experiment. PMID- 18393068 TI - Oxidation and ozonation of waste activated sludge. AB - In this bench-scale study, the treatment of waste activated sludge (WAS) was evaluated using aerobic digestion and ozonation. Two, 2-L batch digesters, one aerated and the other one ozonated, were operated for 30 days in each phase of the study. The aerated digester simulated the aerobic digestion process and served as control to the ozonated digester. In Phase I, the aerated digester was supplied 810 mg O(2) min(- 1), whereas, the ozonated digester was supplied 0.88 mg O(3) min(- 1). In Phase II, the oxygenation rate to the aerobic digester was increased to 1,200 mg O(2) min(- 1) while the ozonation rate was reduced to 0.44 mg O(3) min(- 1). Ozone was more effective than air at oxidizing and reducing both total solids (TS) and volatile solids (VS) in the WAS. TS removals of 50% and 56% were observed for the ozonated digester versus TS removals of 23% and 35% for the aerated digester. VS removals of 40% and 42% were observed for the aerobic digester versus 57% and 74% for the ozonated digester. Aerobic digestion barely met the 38% reduction in VS required by the U.S. Environmental Protection Agency (EPA). The degradation rate constant (K(d)) based on degradable TS for the ozonated digester varied from 0.082 to 0.11 days(- 1) and from 0.067 to 0.09 days(- 1) for the aerobic digester. Total chemical oxygen demand (TCOD) removal in the aerobic digester increased from 30% to 40% from Phase I to Phase II. TCOD removal increased slightly from 57% to 58% in the ozonated digester from Phase I to Phase II. Soluble chemical oxygen demand (SCOD) concentrations in the sludge supernatant increased with digestion time, especially in the ozonated digester. Approximately 0.12 to 0.22 mg SCOD was produced per mg of TS destroyed during ozonation. The specific oxygen uptake rate (SOUR) was consistently below the EPA standard of 1.5 mg O(2) per hr per g TS, indicating that the sludge was well stabilized. The average quantity of oxygen required during aerobic digestion was 1.53 g O(2) per g of TS destroyed. Actual ozone consumption rates were 0.57 mg O(3) per mg TS destroyed and 1.09 mg O(3) per mg TS destroyed for Phase II and Phase I, respectively. PMID- 18393070 TI - Humic substances in bioremediation of industrial wastewater - mitigation of inhibition of activated sludge caused by phenol and formaldehyde. AB - This paper describes studies on the effect of humic substances on bioremediation of industrial wastewater, heavily contaminated with formaldehyde and phenol, using aerobic respirometry. Returned activated sludge (RAS) was the source of biomass. Respirometric data and non-linear regression analysis indicated that the system complied with the Haldane model for inhibitory wastes. It has been found that an addition of humate at the dose of 2000 mg/L substantially reduced this inhibitory effect, resulting in an increase of biological oxygen uptake and in a better removal of both, phenol and formaldehyde. The results show that an application of humic substances to mitigate an inhibition of the activated sludge in wastewater treatment plants may be an attractive alternative to the use of activated carbon or specialized sorbents. PMID- 18393071 TI - Plant germination and growth after exposure to iron cyanide complexes. AB - Phytoremediation has been proposed for treatment of cyanide-contaminated soil. This study was conducted to identify plants with the highest potential for phytoremediation of iron cyanide contaminated soil. Multiple cultivars of two cyanogenic species, sorghum (Sorghum bicolor) and flax (Linum usitatissimum), and one non-cyanogenic species, switchgrass (Panicum virgatum L), were selected for evaluation. The cultivars were screened by quantifying germination and root elongation. Differences in germination emerged among the cultivars (P < 0.05), but these differences appeared to be unrelated to cyanide concentration. The presence of 1000 mg/kg Prussian blue tended to suppress root growth parameters of flax and switchgrass but did not affect sorghum similarly. PMID- 18393072 TI - Modeling nutrient dynamics under critical flow conditions in three tributaries of St. Louis Bay. AB - Previous research results indicated that dry weather condition has complicated impacts on nitrogen dynamics; monitored and modeling data showed both increased and decreased levels. In order to facilitate the total maximum daily loads (TMDLs) development at three tributaries of St. Louis Bay estuary, the nitrogen dynamics were investigated for two designed critical flow conditions by integrating Hydrological Simulation Program Fortran (HSPF), Environmental Fluid Dynamics Code (EFDC), and Water Quality Analysis Simulation Program (WASP). The total amount of precipitation during the dry year corresponded to a flow condition with return period of 50 years, and 10-year return period for wet year. The dry year contributed more total nitrogen (TN) loads per unit flow volume. At the upstream tributaries, the computed peak reach-averaged TN concentrations were significantly higher for dry weather simulation than wet conditions, whereas at the near-bay tributary, there were no significant differences in the peak TN concentrations. Hence, for the upstream tributaries, the nitrogen TMDL calculation should be based on dry weather condition since the decision-makers are more concerned about the worse scenario. PMID- 18393073 TI - Environmental levels of cadmium, lead and mercury in brown hares and their relation to blood metabolic parameters. AB - The purpose of this study was to examine concentrations of selected heavy metals in the liver and kidney of brown hares (Lepus europaeus). In addition, correlations between heavy metals and biochemical parameters in blood plasma were determined. The average concentrations of heavy metals (mmol/L) +/- SD were as follows: liver: Pb 0.221 +/- 0.189, Cd 0.160 +/- 0.140, Hg 0.021 +/- 0.030, kidney: Pb 0.115 +/- 0.125, Cd 1.570 +/- 1.103, Hg 0.030 +/- 0.053. The average concentrations of biochemical parameters in the blood plasma were as follows: Ca 3.16 mmol/L, P 2.19 mmol/L, Mg 1.40 mmol/L, Na 148.71 mmol/L, K 8.12 mmol/L, glucose 6.56 mmol/L, total proteins 56.49 g/L, urea 5.00 mmol/L, total lipids 1.40 g/L, bilirubin 3.97 micro mol/L, cholesterol 1.53 mmol/L, aspartate aminotransferase (AST) 6.06 micro kat/L and alanine aminotransferase (ALT) 1.94 micro kat/L. Average levels of hormones (ng/mL) were as follows: testosterone 2.94, androstendiol 0.13, estradiol 501.59, progesterone 6.63, oxytocin 328.60. Tissue analysis showed an accumulation of lead, cadmium and mercury in the liver and kidney of brown hares. There were no significant correlations between levels of heavy metals in liver, kidney, and biochemical parameters. PMID- 18393074 TI - In vitro copper inhibition of the rabbit spermatozoa motility. AB - Copper is an essential trace element that is strongly bioaccumulated. In this study the effects of this environmental contaminant on various spermatozoa motility parameters in vitro was analyzed. Rabbit spermatozoa were cultured with copper (CuSO(4).5H(2)O) which was added to semen in 5% solution and subsequently diluted 1:1-10. Analysis was carried out using a Computer Assisted Semen Analyzer (CASA) system in 3 time periods (0, 60 and 120 minutes). At Time 0, the highest motility in control group was detected. Motility in groups with copper administration was significantly (P < 0.05) decreased to 8.25-19.37%. The decrease of progressive motility was even more significant - in control 73.37 +/- 7.01% and in experimental groups up to 0%. At Time 60, motility significantly (P < 0.05) decreased from 59.62 +/- 11.40% to 7.50 +/- 3.66%. Progressive motility decreased from 49.62 +/- 16.10% to 0% in the group with the highest copper concentration. After 120 minutes of incubation the motility was 57.75 +/- 5.82% in control group and in all experimental groups it significantly (P < 0.05) decreased to 8.75%. Detailed evaluation of spermatozoa distance (DCL - distance curved line; DAP - distance average path; DSL - distance straight line) and velocity (VCL - velocity curved line; VAP - velocity average path; VSL - velocity straight line) parameters detected significant (P < 0.05) decrease in all studied markers in groups with copper addition in comparison with control group at all time periods. Straightness, linearity, wobble, amplitude of lateral head displacement and beat cross-frequency of spermatozoa were altered weakly. Detected data clearly confirm negative effects of high copper concentrations in semen on spermatozoa motility parameters and subsequent reproductive alteration in male sexual functions. PMID- 18393075 TI - The photo-oxidative destruction of C.I. Basic Yellow 2 using UV/S(2)O(8)(2-) process in an annular photoreactor. AB - The photo-oxidative decolorization of C. I. Basic Yellow 2 (BY2), was investigated using UV radiation in the presence of peroxydisulfate (S(2)O(8)(2-)) in an annular photoreactor at different conditions. S(2)O(8)(2-) and UV-light showed negligible effect when they were used independently. Removal efficiency of BY2 was sensitive to the operational parameters such as initial concentrations of S(2)O(8)(2-), BY2 and pH. The conversion ratios of BY2 at the volumetric flow rates of 330, 500 and 650 mLmin(- 1) were 84%, 78%, 69% in 1 h, respectively. The results showed that in the presence of S(2)O(8)(2-), the photooxidation quantum yield obtained higher than direct photolysis quantum yield, suggesting that photodecay of BY2 was dominated by photooxidation. The electrical energy per order (EE/O) values for decolorization of BY2 solution was calculated. Results show that applying an optimum peroxydisulfate concentration can reduce the EE/O. PMID- 18393076 TI - Impact of harvesting on constructed wetlands performance - a comparison between Scirpus grossus and Typha angustifolia. AB - Three units of free water surface (FWS) constructed wetlands treating domestic wastewater under tropical conditions were examined in terms of water quality and biomass characteristics. One unit (L2) was planted with Scirpus grossus, one with Typha angustifolia (L3), and the unplanted third (L1) served as control. Influent and effluent quality parameters: biological oxygen demand (BOD(5)), nitrate (NO(3)(-)-N), ammonium (NH(4)(+)-N), phosphorus (P), total suspended solids (TSS) and fecal coliforms were regularly measured. The average BOD(5) reductions were 37.0%, 58.5%, and 53.8% for units L1, L2, and L3, respectively. The planted units removed pollutants more effectively although there was no significant difference between the Scirpus grossus and Typha angustifolia units. Plant growth was monitored in marked quadrats by measuring shoot height and other growth parameters. The above-ground organs in L2 and L3 was harvested whenever the shoots reached maximum shoot height and formed flowers. Scirpus grossus had sustainable above-ground biomass production but Typha angustifolia could not sustain repeated harvestings with the above-ground biomass production declining significantly following four consecutive harvests. PMID- 18393078 TI - A validation of self-reported substance use with biochemical testing among patients presenting to the emergency department seeking treatment for backache, headache, and toothache. AB - PURPOSE: This study tests the validity of self-reported illicit substance use against biochemical testing among Emergency Department (ED) patients seeking treatment with narcotics for backache, headache, and toothache and to characterize patients who provide false reports. METHODS: Retrospective chart review comparing the self-reported drug use history obtained during an ED visit during a six-year period (1995-2001) with the results of a biochemical drug screen obtained the same day. RESULTS: 248 patients met screening criteria, 79 (32%) of whom tested positive for unclaimed "drugs of abuse." Patients with a history of "drug abuse" and chronic pain were significantly more likely to test positive for unclaimed drugs than were their counterparts (p=.05 and p<.0001, respectively). No significant difference was found in comparing those with and without multiple ED visits or those requesting a specific narcotic. CONCLUSION: Self-reported drug use is unreliable in this ED subpopulation. When this knowledge is critical for patient care, biochemical testing may be indicated. PMID- 18393079 TI - Posttraumatic stress disorder, gender, and problem profiles in substance dependent patients. AB - Patients with a chronic and severe substance-use disorder who also have a history of posttraumatic stress disorder (PTSD) are thought to have a unique set of problems. The present study assessed psychiatric disorders, psychosocial problems, and traumatic events with structured interviews in 747 men and 693 women enrolling in urban opioid substitution treatment programs from 1995 to 2001. Participants with versus without a history of PTSD were more likely to have a history of many other psychiatric disorders and demonstrated more current and historical medical, employment, family/social, and psychiatric problems. PTSD was generally unrelated to substance-use disorder severity or diagnoses, with the exception of an increased risk of alcohol dependence. Women were more likely than men to have experienced sexual assault, and less likely to have been physically assaulted, although these events precipitated PTSD at equivalent rates across gender. In contrast, witnessing or hearing about the death or injury of others was more likely to precipitate PTSD in women than men. Female gender, exposure to combat, sexual assault, or physical assault, and a history of major mood or anxiety disorder were the best predictors of PTSD in this group. Study limitations are noted. PMID- 18393080 TI - Impact of substance dependence and dual diagnosis on the quality of life of heroin users seeking treatment. AB - In 189 opioid-dependent subjects on methadone maintenance treatment in Barcelona (Spain), we assessed the prevalence of co-occurrence substance use and non substance-use disorders (dual diagnosis) by the Psychiatric Research Interview for Substance and Mental Disorders [PRISM], and the impact on quality of life (HRQoL) by the SF-12 (PCS-12 and MCS-12 scales). Rates of substance and non substance-use diagnoses were 59% and 32%, respectively. Mean scores for PCS-12 and MCS-12 were 44.1+/-10.1 and 39.9+/-11.7, without differences by presence or absence of dual diagnosis. Heroin users on methadone treatment showed a high prevalence of dual diagnosis and a very impaired HRQoL. PMID- 18393081 TI - The role of peers in the initiation and continuation of ecstasy use. AB - This study is a supplement to the Netherlands XTC Toxicity Study (NeXT), funded by grants from the Netherlands Organisation for Health Research and Development as part of its Addiction Programme. To better understand the processes of peer influence and peer selection, in a field study 106 Ecstasy users (67M/39F, average age 25.4 years) were interviewed face-to-face in Amsterdam in 2005. In the initiation of Ecstasy use, peer influence emerged as the dominating mechanism; peer selection was uncommon. In the continuation of Ecstasy use, peer influence and peer selection occurred reciprocally in a dynamic process, although peer influence made a greater relative contribution. Our study confirms that peer influence is a multidimensional process: influence was quite often reciprocal (with respondents both exerting and undergoing influence) and it could have both restraining and encouraging effects on ecstasy use. The study's limitations are noted. PMID- 18393082 TI - A profile of the substance abuse treatment industry: organization, costs, and treatment completion. AB - Nationally representative data from the Alcohol and Drug Services Study, conducted between 1996 and 1999, are used to explore the structure and operation of the substance user treatment industry in the United States. The empirical relationship among client (N=4945) retention and completion, types and use of counseling and medical personnel, diagnostic mix, client demographics, the level of services used, and the cost of treatment in different treatment settings is discussed using tabular presentation and tests of significance. Limitations of the analysis are outlined. This information and analysis are expected to help the research community understand the potential of the ADSS data in addressing many important questions about substance user treatment. PMID- 18393084 TI - Attributions of responsibility for addiction: the effects of gender and type of substance. AB - In 1997, 248 urban university students in central Massachusetts rated responsibility for addiction using the Attributions of Responsibility for Addiction Scale (ARAS), developed for this study with university-based financial support. The vignette-based factorial design varied sex of drug user and type of addictive substance. Factor analysis yielded two subscales: internal and external responsibility attributions; the dependent variable was the internal-to-external attribution ratio. Analysis of variance indicated main effects for substance type and drug abuse experience and showed interaction effects for participant's sex by user's sex and user's sex by substance type. Authors discuss implications, study limitations, and future research. PMID- 18393083 TI - Tobacco, alcohol, and marijuana use among first-year U.S. college students: a time series analysis. AB - The present study sought to evaluate the day-to-day patterns of tobacco, alcohol, and marijuana use among first-year college students in the United States. Using 210 days of weekly time-line follow-back diary data collected in 2002 to 2003, the authors examined within-person patterns of use. The sample was 48% female and 90% Caucasian. Sixty-eight percent of the participants were permanent residents of Indiana. Univariate time series analysis was employed to evaluate behavioral trends for each substance across the academic year and to determine the predictive value of day-to-day substance use. Some of the most common trends included higher levels of substance use at the beginning or end of the academic year. Use on any given day could be predicted best from the amount of corresponding substance use 1 day prior. CONCLUSIONS: Although universal intervention might best be focused in the earliest weeks on campus and at the end of the year when substance use is at its highest, the diversity of substance use trajectories suggests the need for more targeted approaches to intervention. Study limitations are noted. PMID- 18393085 TI - Medical student judgments of adolescents with alcohol use disorders (AUD). AB - The clinical encounter presents opportunities for detection and intervention of adolescent alcohol use disorders (AUDs). AIMS: Investigate (a) identification rate of AUDs, (b) whether AUD identification predicts clinical judgment, and (c) patient characteristics influences on clinical judgment. Medical students (n=123) read a case study and completed questions on diagnosis and clinical judgment. Twenty-five percent of participants identified AUD adolescents, who were more negatively rated than non-AUD adolescents. Prior clinical experience and addiction training predicted AUD identification. Patient race and gender influenced clinical judgment ratings. Addictions training is needed to improve identification rates. Study limitations are noted. PMID- 18393086 TI - The assessment of post-detoxification anhedonia: influence of clinical and psychosocial variables. AB - Anhedonia, or the inability to experience pleasure, may be regarded either as a temperamental trait or as a state symptom of negative schizophrenia or melancholic depression. In substance use disorders, anhedonia was linked to hypoactivity of the dopaminergic mesolimbic system during protracted withdrawal. The authors investigated the influence of recent clinical and social environmental factors on the hedonic capability and related psychopathology in a sample of 70 patients manifesting substance dependence (alcohol, opiates, multiple drugs) without severe comorbidity. Three symptom scales covering anhedonia (Snaith-Hamilton Pleasure Scale, SHAPS; Scale for the Assessment of Negative Symptoms, SANS; Bech-Rafaelsen Melancholia Scale, BRMES) were administered together with the European adaptation of the Addiction Severity Index (EuropASI). The composite scores from the seven areas of the EuropASI were introduced as independent factors in a stepwise regression analysis having symptom scores of anhedonia as dependent variables. Overall, the EuropASI composites do not explain the variability of the scores of anhedonia. Only in few cases, the regression models show a weak predictive capacity for medical status, alcohol use, and drug use composite scores, with R-square values ranging from 10 to 22%. Even if the study's limitations are noted, anhedonia appears as a psychopathological entity independent from other clinical and psychosocial features of treated addicts. PMID- 18393087 TI - Estimating the total mortality among problem drug users. AB - This paper's objective is to develop a method to estimate the total mortality among problem drug users. The total mortality is given by a base rate of mortality not related to drugs and the deaths that are directly and indirectly related to drugs. A fatal poisoning by drugs (overdose) is directly related to drugs, whereas a casualty due to a drug-related disease or a drug-related accident is indirectly related to drugs. As an example of a method to estimate the total mortality, the results from a cohort study among methadone patients in Amsterdam were projected on the whole population of problem drug users in The Netherlands. Due to differences between the problem drug users in Amsterdam and the rest of the country, adjustments were required. It was found that an initial estimation did not require adjustment for injection behavior and gender but did require adjustment for age and the percentage of HIV infection. In a first unadjusted estimation, the total number of deaths among problem drug users in The Netherlands in 2001 was estimated at 606 deaths. After adjustment for age, the estimated mortality decreased to 573 deaths, and after adjustment for HIV infection, this estimation again decreased to 479 deaths. From the ultimately estimated mortality, 11% was considered to be not related to drugs, 23% was attributed directly to drugs, and 66% was attributed indirectly to drugs. The number of direct deaths, as estimated by this method, falls in the same order of magnitude as the number extracted from the Causes of Death Statistics, when selecting cases according to the Drug-Related Deaths Standard as established by the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA). Further cross-validation with other measures will be needed to assess the accuracy of the method, the limitations of which are discussed with respect to stipulating directions for future research. PMID- 18393088 TI - Ascites syndrome in broilers: physiological and nutritional perspectives. AB - Broiler chickens are intensively selected for productive traits. The management of these highly productive animals must be optimal to allow their full genetic potential to be expressed. If this is not done, inefficient production and several metabolic diseases such as ascites become apparent. The causes of the ascites are multifactorial but diet and, particularly, interactions between diet, other environmental and genetic factors play an important role. The relatively high heritability estimates for ascites-related traits and the significance of maternal genetic effects for most of the traits indicate that direct and maternal genetic effects play an important role in development of the ascites syndrome. An imbalance between oxygen supply and the oxygen required to sustain rapid growth rates and high food efficiencies causes ascites in broiler chickens. Because of the relationship to oxygen demand, ascites is affected and/or precipitated by factors such as growth rate, altitude (hypoxia) and environmental temperature. As the high metabolic rate (fast growth) is a major factor contributing to the susceptibility of broilers to ascites, early-age feed or nutrient restriction (qualitative or quantitative) or light restriction in order to slow down the growth rate seem practically viable methods, since final body weight is not compromised. Manipulation of the diet composition and/or feed allocation system can have a major effect on the incidence of ascites. Optimization of the house temperature and ventilation in cold weather seem helpful practices to decrease ascites incidence. PMID- 18393089 TI - A recombinant avian leukosis virus associated with fowl glioma in layer chickens in Japan. AB - Fowl glioma is characterized by multiple nodular growth of astrocytes, and fowl glioma-inducing virus belonging to avian leukosis virus has been isolated from Japanese bantam as a causal agent. Subcutaneous neoplasms of the head and neck have been reported in layer chickens since 2003 in Japan, and fowl glioma concurred in these affected layers. In the present study, the histopathology of 240 layers, including 18 layers with subcutaneous neoplasms and 222 layers kept with the affected layers, was performed to clarify the characteristics of fowl glioma in layers. Microscopically, 103 layers showed non-suppurative encephalitis, and four layers had locally extensive proliferation or multiple nodules of astrocytes. Gliomas concurred in 11 layers with subcutaneous neoplasms and occurred independently in three layers. In addition, two layers had locally extensive proliferation of small, round cells in the cerebrum. The fowl glioma inducing virus genome was not detected in the affected brains by nested polymerase chain reaction. Ten isolates were obtained from the affected brains. By nucleotide sequencing of the env gene, SU coding regions of these isolates were most closely related to myeloblastosis-associated virus-like viruses, but TM regions showed the highest similarity to endogenous viral (ev) loci. The genome of one isolate mainly consisted of ev loci and contained several parts of other avian leukosis/sarcoma viruses. These results show that the causal avian leukosis virus of fowl glioma is not just fowl glioma-inducing virus and that different avian leukosis virus strains having oncogenicity in the central nervous system by recombination are spread in layers in Japan. PMID- 18393090 TI - Oviductal volvulus in a Nera black chicken (Gallus gallus domesticus) in Nigeria. AB - A case of oviductal volvulus, a rare disorder with oviductal cyst as a possible aetiology, is described in an 11-month-old Nera black chicken (Gallus gallus domesticus). The condition was observed during routine postmortem examination of 550 carcasses from a flock of 16,100 birds between September 2005 and June 2006. The affected portion of the oviduct and the cyst twisted almost 360 degrees clockwise around the dorsal ligament with severe congestion, hyperaemia, oedema, dilatation and devitalization. This is the first reported case of oviductal volvulus in a domestic chicken. PMID- 18393091 TI - Teratoma in the cerebrum of a fantail pigeon. AB - This is the first report of a primary teratoma in the cerebral cortex of a 1-year old fantail pigeon and one of the few reports of intracranial teratomas in birds. The clinical signs were sudden onset of listlessness and a head tilt to the right. The right cerebral hemisphere contained an unencapsulated teratoma that included adipose, cartilaginous, fibrous and undifferentiated mesenchymal tissue as well as keratinized and glandular epithelial structures. Immunohistochemistry designed for mammals proved very useful and has been used to investigate the two germ cell lines, epithelial and mesenchymal, detected in the neoplasm. Indirect immunohistochemistry testing using vimentin, pancytokeratin, smooth muscle actin, neuron-specific enolase, and S-100 was done. Vimentin, smooth muscle actin and pancytokeratin immunoreactivity was strong. Neuron-specific enolase immunoreactivity was strongly positive in the normal brain adjacent to the neoplasm but there was no immunoreactivity within the neoplasm. Also, there was no S-100 immunoreactivity, suggesting that the mammalian proteins on which the immunohistochemistry is based are not present in pigeons. PMID- 18393092 TI - Comparison of three template preparation methods for routine detection of beak and feather disease virus and avian polyomavirus with single and nested polymerase chain reaction in clinical specimens. AB - Direct comparisons are important when assessing the value of DNA extraction methods for diagnostic virology as the inhibitors present and the efficiency of extraction vary with the target infectious agent as well as the species and the site from which the clinical sample was obtained. Three DNA extraction methods were compared for routine polymerase chain reaction (PCR) detection of beak and feather disease virus (BFDV) in whole blood and feather samples and of avian polyomavirus (APV) in feather samples. Boiling in Chelex 100 Resin was found to be the most sensitive method for detection of BFDV or APV DNA in both feather and blood samples. In combination with nested PCR it enabled detection of BFDV DNA in 13/13 positive whole blood samples and in 22/23 positive feather samples. It also enabled detection of APV in 31/31 samples detected as positive in this study. NucleoSpin kits enabled detection of BFDV DNA in only 9/13 blood samples and in 18/23 feather samples. The lower rate of BFDV DNA detection when using NucleoSpin kits was not a result of inhibition of PCR in most cases. The NucleoSpin Tissue Kit enabled detection of APV DNA in 29/31 feather samples. Inhibition of DNA amplification was observed when using the DNAzol Direct kit. Therefore, of the methods evaluated here, Chelex 100 Resin treatment of samples was the best option for routine testing for BFDV and APV DNA in clinical samples. PMID- 18393093 TI - Attenuation of very virulent infectious bursal disease virus and comparison of full sequences of virulent and attenuated strains. AB - A very virulent strain of infectious bursal disease virus (IBDVks) was isolated from the bursae of Fabricius of IBDV-affected broiler chickens. Following 43 serial passages in specific pathogen-free embryonated eggs, an attenuated strain was established (IBDVmb). Dosages of IBDVmb in the range 10(2) to 10(4) embryo infective dose of 50% were found to be safe and protective for commercial chicks. Chickens vaccinated with live vaccine containing IBDVmb responded with precipitating and type-specific neutralizing antibodies, and were immune to subsequent challenge with a very virulent IBDV. IBDVmb has been used as an attenuated vaccine throughout the world since 1993. A comparison of the full sequences of the virulent and attenuated strains (IBDVks and IBDVmb, respectively) revealed seven nucleotides that were different, four of them leading to changes in the amino-acid sequence. Comparison of the protein sequence of these strains and published sequences of very virulent and attenuated phenotypes lead us to suggest that the novel difference responsible for virulence of the Israeli strains are: residue 272 (VP2, very conserved site) and residue 527 (VP4), both in segment A, and in segment B (VP1) residues 96 and 161 (both conserved). Our study strengthens the possibility that more than one protein is involved in IBDV attenuation. In all reports, including ours, virulence was reduced without affecting antigenicity of the neutralizing epitopes in VP2. This could have practical implications for attenuated-vaccine development. PMID- 18393094 TI - Coccidial infections in commercial broilers: epidemiological aspects and comparison of Eimeria species identification by morphometric and polymerase chain reaction techniques. AB - The objective of this study was to add to existing knowledge of the epidemiology and the aetiology of coccidial infections in commercial broiler flocks. Polymerase chain reaction (PCR) and morphometric identification of the Eimeria species were compared as means of differentiation in the field samples of faeces and litter. For morphometry, the Eimeria species were categorized into three groups based on lengths of the oocysts. Two random samples of commercial broilers were studied, one during 2000/01 and the other during 2003/04. The prophylactic regime (in-feed narasin), husbandry and methods applied were broadly the same for both subpopulations. Coccidial infection prevalence increased from approximately 45% to approximately 75% during this period, but infection levels (oocysts per gram of faeces) did not significantly change. There were substantial geographical differences in both prevalence and infection levels. A change in Eimeria species profile occurred during the study period. Five Eimeria species were identified at slaughter, by PCR targeting the ITS-1 region of the genome; Eimeria acervulina (100%), Eimeria tenella (77%), Eimeria maxima (25%), Eimeria praecox (10%) and Eimeria necatrix (2%). PCR and morphometric tentative identification were in complete agreement in only 49% of the cases. PMID- 18393095 TI - Differential diagnosis between type-specific duck hepatitis virus type 1 (DHV-1) and recent Korean DHV-1-like isolates using a multiplex polymerase chain reaction. AB - Duck hepatitis can be caused by four types of viruses: duck hepatitis virus (DHV) type 1 (DHV-1), DHV-1a (a variant strain of DHV-1), DHV-2 and DHV-3. In Korea, duck hepatitis has been associated with two types of DHV-1, original DHV-1 type specific strain (DHV-1s) and the recently reported DHV-1 variant strains (DHV 1v). The pathogenicity and pathological findings of ducklings infected with the recent DHV-1v isolates, AP-04114 and AP-04203, were almost identical to those infected with members of the DHV-1s, DHV-HS and the type-specific strain DRL-62. To be able to monitor the epidemiological patterns exhibited by the two Korean types, a specific gene-based differential diagnostic method based on multiplex polymerase chain reaction was developed. The primers selected were designed to bind to and amplify conserved regions within the RNA-dependent RNA polymerase (3D) gene, the complete capsid (P1) region or the 5'-untranslated region to distinguish between the DHV-1s and DHV-1v groups. The described multiplex polymerase chain reaction method was able to selectively recognize ducklings infected with either of the two groups of Korean isolates. The method was also able to distinguish between DHVs and other avian-originated RNA viruses. The detection limit of the diagnostic method was determined to correspond to 10(3) copies viral RNA and 100 pg used as starting template. As a result, the use of this test allows rapid and early diagnosis of two different virus types affecting the commercial duckling industry. PMID- 18393096 TI - Development of a polymerase chain reaction assay for specific identification of Clostridium colinum. AB - Clostridium colinum is the causative agent of ulcerative enteritis, a serious disease of the bobwhite quail (Colinus virginianus) and sporadically of young chickens. The aim of the present study was to develop a polymerase chain reaction (PCR) assay specific for C. colinum identification. The 16S rDNA sequence of C. colinum was analysed and two species-specific primers were designed. The specificity of these primers was tested with closely related Clostridium species and the expected amplified product (935 base pairs) was observed only with DNA from samples containing C. colinum. Results from performing PCR assays on faecal samples from quails spiked with different concentrations of C. colinum, showed that the detection limit of the assay was 1.6 x 10(4) colony-forming units per gram of faecal material. This PCR assay can be used in diagnostic laboratories to confirm the presence of C. colinum in pure cultures and could be used to screen enriched samples or faecal samples for the presence of this pathogen. PMID- 18393097 TI - The vaccination-challenge trial: the gold standard test to evaluate the protective efficacy of infectious coryza vaccines. AB - Infectious coryza is an upper respiratory tract disease of chickens with the major impact occurring in multi-age flocks. We investigated the relationship between the level of antibodies, as detected by a haemagglutination-inhibition (HI) assay, in infectious coryza-vaccinated chickens and the protection against challenge in those chickens. In one experiment, chickens given a single dose of either of two infectious coryza vaccines lacked a detectable HI response to vaccination but showed significant levels of protection 11 weeks after vaccination. In contrast, in chickens given two doses of an infectious coryza vaccine and challenged 3 weeks after the second vaccine dose, there was a strong serological response with 36/40 birds having a HI titre of 1/20 or greater. In this trial there was an apparent relationship between titre and subsequent protection, with none of the 32 chickens with a titre of 1/40 or 1/80 showing any clinical signs and only one of the same group yielding the challenge organism on culture. In contrast, three of the four vaccinated chickens with a HI titre less than 1/5 developed the typical clinical signs of coryza and yielded the challenge organism on culture. Overall, our results suggest that HI titres cannot be regarded as a definitive predictor of vaccine efficacy. We suggest that the vaccination-challenge trial is the gold standard for the evaluation of the immune response to infectious coryza vaccines. PMID- 18393098 TI - Development of a non-radioactive digoxigenin cDNA probe for the detection of avian encephalomyelitis virus. AB - A non-radioactive digoxigenin cDNA probe for detection of avian encephalomyelitis virus (AEV) was developed. The cDNA probe hybridized specifically with a fragment of VP1 gene of AEV genome was found to be sensitive with as little as 10 pg target DNA fragment in a sensitivity test. Infected chicken embryos were strongly labelled by testing the probe against a range of AEV strains, and no non-specific reaction was observed in non-infected chicken embryos as well as five other avian pathogenic virus-infected samples used as negative controls. Furthermore, the cDNA probe was capable of detecting AEV from chicken embryo brain at 3 days post inoculation as compared with an immunofluorescence assay, which required up to 5 days of incubation in the embryos. In clinical application, five out of 16 clinical brain samples that were negative by the immunofluorescence assay were positive for AEV by the cDNA probe. Both the sensitivity and specificity of the developed cDNA probe indicated that it is a highly promising and reliable diagnostic tool for the detection of AEV infections. PMID- 18393099 TI - Pathogenesis of type 2 turkey astroviruses with variant capsid genes in 2-day-old specific pathogen free poults. AB - The pathogenicity of three different type 2 turkey astroviruses (TAstV-2) was studied in specific pathogen free turkeys. These viruses differ based on sequence analysis of the capsid gene. Poults were inoculated at 2 days of age and examined during 14 days for clinical signs and virus shedding. All inoculated poults presented signs of enteric disease including diarrhoea and growth depression. Virus presence and shedding was detected by real-time reverse transcriptase polymerase chain reaction from intestinal contents and cloacal swabs collected at 3, 7 and 14 days post-inoculation. Viraemia was also confirmed by this method. Common lesions observed at necropsy were dehydration; distended intestines filled with watery contents and undigested feed, and dilated caeca with foamy contents. Microscopic lesions present in the intestines consisted of mild crypt hyperplasia, villous atrophy and lymphocytic infiltration, and were most common in the jejunum. Presence of the viruses was demonstrated by immunohistochemistry and by in situ hybridization in both villi and crypt enterocytes in the jejunum and, less frequently, the duodenum, ileum and caeca. Mild lesions consisting mainly of lymphocytic infiltration were also observed in other organs including the pancreas, liver, spleen and kidneys. Mild to moderate bursal atrophy occurred in all TAstV-2-infected poults examined; however, no specific viral staining was observed in this organ or any other tissues examined apart from the intestines. In conclusion, TAstV-2 viruses with variant capsids produce a similar enteric disease in young turkeys and may also affect the immune system of the birds by causing bursal lymphoid depletion. PMID- 18393101 TI - The success of bariatric surgery as treatment of type 2 diabetes should be a call to revitalize anti-obesity drug discovery and development. PMID- 18393102 TI - Recent considerations in the treatment of hypothyroidism. AB - Thyroid hormone deficiency has been recognized and treated with various forms of thyroid hormone replacement over the last century. Since the 1950s, synthetic L thyroxine has been the therapy of choice. However, there is now recognition that the currently available regimens for the treatment of hypothyroidism may not adequately address the needs of all patients. This review summarizes recent considerations in the field of thyroidology to address the potential for improvement in the treatment of patients. The goal of these improvements should be to achieve both clinical and chemical euthyroidism. PMID- 18393103 TI - New approaches to treatment of secondary hyperparathyroidism. AB - A large number of patients with chronic kidney disease are affected by secondary hyperparathyroidism. The related mineral metabolism abnormalities are associated with an increased relative risk of morbidity and mortality. The management of secondary hyperparathyroidism is made more complex by the fact that the disease progresses over time. Recent approaches to its management include vitamin D analogs, non-calcium/non-aluminum containing phosphate binders and calcimimetics. PMID- 18393104 TI - Sirtuins: novel targets for metabolic disease. AB - Sirtuins represent a novel family of enzymes that are collectively well situated to help regulate nutrient sensing and utilization, metabolic rate and ultimately metabolic disease. Activation of one of these enzymes, SIRT1, leads to enhanced activity of multiple proteins, including peroxisome-proliferator activated receptor coactivator-1alpha (PGC-1alpha), which helps to mediate some of the in vitro and in vivo effects of sirtuins. As such, enhanced SIRT1 activity decreases glucose levels, improves insulin sensitivity, increases mitochondrial number and function, decreases adiposity, improves exercise tolerance and potentially lowers body weight. SRT-501 is a proprietary formulation of resveratrol with improved bioavailability. As such, SRT-501 represents the first in a novel class of SIRT1 activators that has proven to be safe and well-tolerated in humans. Clinical trials in type 2 diabetic patients are currently underway. PMID- 18393105 TI - Advances in glycogen phosphorylase inhibitor design. AB - The regulation of glycogen metabolism is a major therapeutic strategy for blood glucose control in type 2 diabetes. Because glycogen phosphorylase catalyzes the first step in the phosphorolysis of glycogen, it has become a potential key target for controlling hyperglycemia in this disease. This review focuses on advances in new, mostly synthetic, molecules that inhibit glycogen phosphorylase, and describes progress in our understanding of the mechanism of action of these inhibitors gained through X-ray crystallographic studies. PMID- 18393106 TI - Growth hormone use in the treatment of idiopathic short stature. AB - Human growth hormone (hGH) therapy has been used for the possible improvement of adult height in individuals with idiopathic short stature (ISS) for more than 20 years. However, given its heterogeneity, and the distinction between 'partial' or 'transient' GH deficiency, the exact definition of ISS is difficult. Since recombinant hGH became available, individuals with all types of GH deficiency have been extensively treated, often without the need for diagnostic justifications. On the other hand, for ISS individuals GH treatment has so far only been possible in a few countries, or in the context of clinical trials. If hGH is certainly effective in individuals with severe GH deficiency, its efficacy is lower and with a high individual-to-individual variability of response in those with non-severe GH deficiency and with ISS. The most important variables associated with a favorable growth response to hGH therapy in GH- and non-GH deficient individuals are first-year growth response, younger age at start of treatment, difference at start from target height standard deviation score, GH dose, and other variables not necessarily associated with GH peak levels after provocative stimuli. A better choice of individuals to be treated successfully will improve the cost-effectiveness of the treatment, making it more acceptable to the scientific community and other stakeholders. PMID- 18393107 TI - Alogliptin, a potent and selective dipeptidyl peptidase-IV inhibitor for the treatment of type 2 diabetes. AB - Takeda San Diego Inc is developing alogliptin, a small-molecule, orally available dipeptidyl peptidase IV (DPP IV) inhibitor, for the potential treatment of type 2 diabetes. In January 2008, Takeda announced that an NDA for alogliptin had been submitted to the FDA. PMID- 18393108 TI - Cetilistat, a new lipase inhibitor for the treatment of obesity. AB - Alizyme plc, in collaboration with Takeda Pharmaceutical Co Ltd in Japan, is developing cetilistat, an oral non-absorbed synthetic lipase inhibitor, derived from Alizyme's pancreatic lipase inhibitor research program, for the potential treatment of obesity in patients with or without diabetes. It is also under investigation for the potential management of type 2 diabetes. Several phase II clinical trials for obesity have been completed. Phase III trials were in preparation at the time of publication. PMID- 18393109 TI - Pegloticase, a polyethylene glycol conjugate of uricase for the potential intravenous treatment of gout. AB - Savient Pharmaceuticals Inc (formerly Bio-Technology General Corp), under license from Duke University, is developing pegloticase, PEG conjugates of uricase (urate oxidase), for the potential treatment of gout. The in-life portion of the phase III trials have been completed. PMID- 18393110 TI - Early and late outcomes of percutaneous transluminal angioplasty of cephalad arteries. AB - BACKGROUND: Efficacy of carotid endarterectomy (CEA) in prevention of stroke in patients with carotid artery stenosis has been confirmed in randomised trials. Carotid artery stenting (CAS) is a routine clinical practice and recent results of CAS are not worse than CEA. Moreover, percutaneous transluminal angioplasty (PTA) techniques allow other cephalad arteries to be dilated. AIM: To assess early and long-term outcome of PTA of cephalad arteries and to determine risk factors of early and late major adverse cardiovascular and cerebral events (MACCE). METHODS: The study group consisted of 223 consecutive patients (151 males, 67.7%, mean age 65.3+/-8.6) in whom 256 PTA procedures of cephalad arteries were performed. Two hundred and forty-two internal carotid, 7 common carotid and 15 vertebral arteries were dilated. Thirty-four patients underwent one-stage carotid and coronary procedures, while in 46 patients one-stage carotid and peripheral procedures were performed. Neuroprotection with a distal protection device was used in 51.5% of cases. The procedures were divided into two groups: with high (n=181) and low (n=75) risk of cardiovascular events. Early and late events were recorded and analysed subsequently. RESULTS: In hospital 30 day MACCE occurred in 12 (4.6%) patients, including 7 (2.7%) strokes, 3 (1.1%) myocardial infarctions and two (0.8%) deaths. Transient ischaemic attacks were observed in 8 patients, pulmonary oedema in 3 cases, as well as a single episode of retinal artery embolisation and acute renal insufficiency. The incidence of 30 day MACCE was not significantly higher in the high-risk group (6.07 vs. 1.33%; NS), but the risk of any adverse event was significantly higher (p=0.03). There was no difference in stroke incidence between procedures with or without neuroprotection (2.27 vs. 3.22%; NS). There was no difference in risk of MACCE between angioplasty of cephalad artery and one-stage cephalad and coronary artery angioplasty procedure (3.6 vs. 5.5%; NS). During 50.3+/-20 months of follow-up there were 16 (7.1%) deaths, 9 (3.5%) strokes and 6 (2.3%) re-stenoses confirmed angiographically. One-year total survival and one-year MACCE-free survival rates according to the Kaplan-Meier analysis were 94.9% and 89.0%, showing a trend towards better outcome in the low-risk group (F-Cox=2.46; p=0.19 and F-Cox=2.17; p=0.09 respectively). CONCLUSIONS: Percutaneous transluminal angioplasty of cephalad arteries is safe and feasible, with a low periprocedural complication rate and good late outcome. Carotid artery stenting is an alternative method to CEA. PMID- 18393111 TI - The prevalence of potentially unstable coronary lesions in patients with coronary artery disease--virtual histology study. AB - BACKGROUND: Histopathological studies indicate that coronary artery lesions with a thin fibrous cap and large necrotic core (thin-cap fibroatheromas, TCFA) are characterised by a high risk of rupture and can potentially trigger acute coronary syndrome (ACS). Atherosclerotic lesions with a well preserved fibrous cap (fibroatheromas, FA) are considered to be more stable ones. Intravascular ultrasound virtual histology (IVUS-VH) enables identification of FA and TCFA in vivo. There are no published data regarding IVUS-VH derived occurrence of both FA and TCFA in patients with different clinical presentation. AIM: To determine IVUS VH derived occurrence of FA and TCFA in coronary arteries of patients with chronic stable angina in comparison with recent or acute ACS subjects. METHODS: Intravascular ultrasound examination was performed in 60 patients, who were prospectively enrolled to three groups: group A--chronic stable angina, without a history of ACS within the previous 12 months; group B--recent ACS (4 weeks-3 months); group C--acute phase of ACS. RESULTS: The final analysis included 75 non culprit lesions (group A: n=29 lesions; group B: n=22; group C: n=24). There were no significant differences in lesions' angiographic and ultrasound characteristics between the studied groups. There was no significant difference in the occurrence of FA lesions between the studied groups (20.7 vs. 22.7 vs. 4.2, respectively; NS). There was a significant difference in TCFA occurrence between the studied groups (31.0 vs. 50.0 vs. 79.2%; p <0.01). CONCLUSIONS: The present study confirms higher occurrence of thin-cap fibroatheromas in patients with clinically confirmed coronary instability. It also indicates that IVUS-VH can be a valuable tool for rupture prone lesion identification, which might help in better risk stratification in coronary artery disease patients. PMID- 18393112 TI - Heart-type fatty acid binding protein--a reliable marker of myocardial necrosis in a heterogeneous group of patients with acute coronary syndrome without persistent ST elevation. AB - BACKGROUND: Myocardial infarction (MI) is one of the most serious challenges of contemporary cardiology. Among biochemical markers, heart-type specific fatty acid binding protein (h-FABP) has a high potential as a marker for the early diagnosis of acute MI. The h-FABP is released early to the bloodstream and may be useful for both rapid confirmation and exclusion of infarction. As opposed to patients with ST segment elevation MI (STEMI), patients with unstable angina (UA)/non-ST segment elevation MI (NSTEMI) present a heterogeneous group in which the confirmation of MI often meets diagnostic difficulties. A rapid, qualitative immunoenzymatic 'point of care' type test, revealing h-FABP in blood, has recently been made available (CardioDetect med). AIM: To evaluate diagnostic value of early measurements of h-FABP and other markers of necrosis (cTnT, CK-MB, CK-MB mass) in a group of 100 patients with an acute coronary syndrome (ACS) without persistent ST segment elevation (NSTE ACS). METHODS: We studied 100 consecutive patients (34 women, 66 men; mean age 61.6 years) with strong suspicion of NSTE ACS and chest pain lasting <24 h before admission. During admission and after 3 and 6 hours patients had measured a panel of conventional biomarkers as well as quantitative measurements of h-FABP (on admission and 3 hours later) using CardioDetect med. The ultimate diagnosis of infarction (NSTEMI) was confirmed in case of a second (6 h after admission) positive quantitative result of cardiac troponin. Non-ST segment elevation MI was finally diagnosed in 56 patients. RESULTS: The comparison of diagnostic utility of all analysed biomarkers of necrosis revealed that h-FABP was superior to other parameters, when measured on admission, and was characterised by 94.7% sensitivity, 100% specificity, 100% positive predictive value, 93.4% negative predictive value and 97% accuracy. Other biomarkers had on admission lower sensitivity - 70.1% for CK-MB mass, 66.7% for CK-MB, 64.9% for cTnT, whereas their specificity was 97.6% for CK-MB mass, 93% for CK-MB and 100% for cTnT. CONCLUSIONS: Qualitative h-FABP test (CardioDetect med) showed excellent sensitivity, higher than measurements of CK-MB mass, CK-MB, and cTnT on hospital admission, and high specificity in the patient group with NSTE ACS. The h-FABP seems to be an excellent biochemical cardiac marker for diagnosing NSTEMI, especially in its early phase, allowing exclusion of myocardial necrosis. PMID- 18393113 TI - Association between retinopathy, microalbuminuria and coronary perfusion in young patients with type 1 diabetes mellitus. AB - BACKGROUND: In patients with type 1 diabetes mellitus (DM) impairment of the coronary circulation has been observed. This phenomenon could be ascribed to the existence of a specific cardiomyopathy. Disturbances in other microcirculation beds--renal and ocular--are mirrored by microalbuminuria and retinopathy, respectively. The association between coronary microvascular dysfunction and the presence of microalbuminuria and retinopathy is not clear. Recognition of the interrelationships between microalbuminuria, retinopathy and the impairment of coronary circulation could allow for a simple estimation of coronary perfusion in these patients. AIM: To assess coronary blood flow velocity in young patients with type 1 DM using transoesophageal Doppler echocardiography with dipyridamole and to analyse the possible relationship between the impairment of coronary flow and retinopathy as well as microalbuminuria. METHODS: The study group consisted of 36 patients, aged from 18 to 35 (mean: 25+/-5) years with type 1 DM lasting from 8 to 27 years. Diabetes was the only disease and none of the patients had any history of cardiovascular diseases or any abnormalities in physical examination. The control group consisted of 23 age-matched healthy volunteers. All subjects underwent transoesophageal echocardiography with dipyridamole to assess coronary flow velocity reserve (CFVR). RESULTS: In the study group CFVR and maximal flow velocity after dipyridamole were significantly decreased (2.4+/ 0.6 vs. 3.4+/-0.7; p <0.001 and 125.7+/-31.4 vs. 168.00+/-12.9 cm/s; p <0.001, respectively). The basal flow velocity was comparable in both groups (55.9+/-14.6 vs. 52.2+/-11.6 cm/s; p=0.32). Decrease in CFVR in the study group was associated with a smaller increase in coronary flow velocity after dipyridamole challenge. There was no relationship between coexisting microalbuminuria, retinopathy and the CFVR values. CONCLUSIONS: In young patients with type 1 DM, without any clinical cardiovascular abnormalities, decreased coronary perfusion is observed. The presence of microalbuminuria or retinopathy is not associated with the alterations in coronary perfusion. PMID- 18393114 TI - Assessment of left and right ventricular diastolic function in patients with systemic sclerosis. AB - BACKGROUND: Systemic sclerosis (SSc) is a connective tissue disease characterised by vascular changes and immunologically induced fibrosis of the skin and internal organs. Systemic sclerosis may be associated with both right (RV) and left ventricular (LV) diastolic dysfunction. AIM: To analyse RV and LV myocardial diastolic function in patients with SSc and its relation to exercise capacity. METHODS: We prospectively studied 51 consecutive patients (47 females, 4 males, age 53.3+/-15.2 years) with SSc (mean disease duration 9+/-12.4 years) and a group of 31 age-matched healthy subjects (28 females, 3 males, age 52.68+/-12.1 years). In addition to conventional investigation, transthoracic echocardiography (TTE) for assessment of RV and LV myocardial diastolic function and 6-minute walking test (6MWT) were performed. RESULTS: Abnormal LV filling, as expressed by an inverted mitral E/A ratio (Mit E/A <1), was detected in 28 (55%) SSc patients and in 8 (25%) controls (p <0.001). The mean value of Mit E/A in the SSc group was lower than in controls (1.0+/-0.3 vs. 1.2+/-0.3, p=0.04). There were no differences in pulmonary venous flow between SSc patients and controls. The mean value of Tei index for the LV was higher in SSc than in controls (0.44+/-0.08 vs. 0.38+/-0.05, p <0.001). Abnormal RV filling, as expressed by an inverted tricuspid E/A ratio (TR E/A <1), was detected in 16 (31%) SSc patients and in 5 (16%) controls (p <0.001). Patients with SSc were found to have an inverted Tr E/A ratio (Tr E/A <1), indicating abnormal RV filling. The mean value of Tr E/A in SSc was lower than in controls (1.0+/-0.2 vs. 1.2+/-0.3, p=0.04). The mean value of Tei index for the RV was higher in SSc patients than in controls (0.35+/ 0.07 vs. 0.29+/-0.03, p <0.001). In multiple regression analysis Tr E/A ratio was independently correlated with Mit E/A ratio (r=0.65, p=0.01). The mean 6MWT distance was shorter in the SSc group than in controls (528+/-100.6 vs. 617.7+/ 80 m, p <0.001) and the mean saturation of capillary blood after the 6-MWT was lower in SSc patients (92.7+/-4.9 vs. 97.2+/-1.2%, p <0.001). Mean desaturation after test and D sat was significantly more pronounced in the SSc group than in controls (3.4+/-3.1 vs. 0.7+/-0.9, p <0.001). The Tr E/A ratio and Mit E/A ratio correlated positively with 6MWT distance (r=0,49, p=0.01 and r=0.48, p=0.02). CONCLUSIONS: Impaired RV and LV relaxation is observed in a significant percentage of SSc patients and is associated with decreased exercise capacity. PMID- 18393115 TI - [Morphological and clinical aspects of Danon disease]. AB - BACKGROUND: Danon disease, an X-linked hypertrophic cardiomyopathy, is caused by primary deficiency of lysosome-associated membrane protein (LAMP-2). The pathological hallmark of the disease is the appearance of intracytoplasmic vacuoles containing autophagic material and the absence of LAMP-2 activity in the muscle. AIM: To define the LAMP-2 protein deficiency we investigated cardiac and skeletal muscle of a 19-year-old man with hypertrophic cardiomyopathy without clinically apparent skeletal myopathy or mental impairment, whose mother died suddenly at 46 years of age. METHODS: Clinical, morphological, immunohistochemical and ultrastructural analysis was performed. Paraffin sections of cardiac muscle were stained using routine histochemical methods. Frozen sections of skeletal muscle were stained using histochemical methods as well as using monoclonal antisera against N-terminal of dystrophin and antisera against LAMP-2. Ultrastructural examination of both cardiac and skeletal muscle specimens were performed. RESULTS: Cardiac and skeletal muscle revealed an excessive accumulation of early and late autophagic vacuoles containing various cytoplasmic debris. In immunohistochemical analysis the vacuolar membrane seen in skeletal muscle was decorated with antibody against dystrophin and such vacuoles were negative for LAMP-2. CONCLUSION: Ultrastructural and immunohistochemical analysis of skeletal muscle (less invasive than myocardial biopsy) may be used in diagnosis of Danon disease. Early diagnosis of Danon disease is important for timely cardiac transplantation, the only effective therapeutic option. PMID- 18393116 TI - [Short QT syndrome--a case report]. AB - We present two new cases of the short QT syndrome--a 23-year-old male and his 42 year-old mother. Invasive electrophysiological study was negative in both patients. Due to polymorphic ventricular tachycardia recorded during Holter ECG monitoring, an ICD was implanted in the male patient for primary prevention. Blood samples were collected for further genetic studies. Diagnosis and management of patients with the short QT syndrome are discussed. PMID- 18393117 TI - [Acute coronary syndrome in the form of variant angina pectoris while performing stress echocardiography]. AB - A case of a 57-year-old female who underwent dobutamine stress echocardiography is presented. During the test, acute ischaemia with contractile abnormalities and profound hypotension occurred. Coronary angiography showed no abnormalities and coronary spasm was suspected as a possible cause of symptoms. PMID- 18393118 TI - [Late thrombosis after bare metal stent implantation in a patient with acute coronary syndrome]. AB - We present a case of a 47-year-old man with ST-segment-elevation myocardial infarction (STEMI), successfully treated with bare metal stent implantation. After 20 months the patient was readmitted with another episode of STEMI due to in-stent thrombosis following voluntary withdrawal of anti-platelet therapy. The implications of late stent thrombosis after bare metal stent implantation are discussed. We compare this phenomenon with thrombosis after implantation of drug eluting stents. Various aspects and potential causes of late stent thrombosis are reviewed, especially the issues pertaining the use of anti-platelet therapy and duration of this therapy. PMID- 18393119 TI - [Acute ischaemic stroke in a young male with patent foramen ovale]. AB - We present a case of a 35-year old patient with a patent foramen ovale and acute ischaemic stroke. The patient underwent successful percutaneous closure of foramen ovale with an Amplatzer device. Epidemiology, diagnosis and treatment of this condition are discussed. PMID- 18393120 TI - [Application of bone marrow cells in myocardial infarction]. PMID- 18393121 TI - [Laminopathies--interdisciplinary problem]. PMID- 18393122 TI - Pseudo-Wellens syndrome--a case report. PMID- 18393123 TI - [Recurrent rupture of a sinus of Valsalva aneurysm in a patient with a bicuspid aortic valve treated with percutaneous implantation of an Amplatzer occluder]. PMID- 18393124 TI - [Invasive cardiologist with radiologist cooperation using coronary angio-CT during revascularisation of an angiographically difficult patient with ST-segment elevation myocardial infarction--a case report]. PMID- 18393125 TI - [Ablation of right ventricular tachycardia with a bizarre QRS complex--a case report]. AB - We describe a case of a 77-year-old patient with recurrent, symptomatic ventricular tachycardia with a bizarre QRS complex originating from the right ventricle lateral wall. The small region of slow conduction was the critical substrate for initiation and maintenance of ventricular tachycardia, confirmed by a successful one RF application. PMID- 18393126 TI - [Endovascular treatment of aneurysm of thoracic aorta (TEVAR) in 2008--for whom, why, when?]. PMID- 18393128 TI - The prevalence of RET/PTC mutations in papillary thyroid cancers in Turkish population and its relation between tumor histopathology and prognostic factors. AB - OBJECTIVE: In recent years, thyroid cancer has been at the forefront of molecular pathology as a result of the consequences of the Chernobyl disaster and the recognition of the role of RET/PTC rearrangements in papillary thyroid carcinomas (PTCs). Correlation of RET/PTC expression with clinical outcome is controversial. This study aims to identify the prevalence of RET/PTC oncogene expression in Turkey, and to investigate the correlation between RET/PTC oncogene expression and the known prognostic factors of PTC in 101 patients. METHODS: The RET rearrangements were examined by means of reverse transcriptase-polymerase chain reaction analysis, with primers flanking the chimeric region. Statistical evaluation was performed by using Independent samples t-test, One-sample Chi square test and Pearson Chi-square or Fisher's Exact Test. RESULTS: RET/PTC was determined positive in 67(66.3%) of totally 101 patients (p<0.001). RET/PTC1 in 32(31.7%), RET/PTC3 in 21(20.8%), RET/PTC1+RET/PTC3 both in 10(9.9%) patients were found to be positive. There was RET/PTC2 positiveness in two patients, RET/PTC2,3 positiveness in one patient, and RET/PTC1,2,3 positiveness in one patient. No statistical difference was found between RET/PTC1 and RET/PTC3. None of genetico-clinical analyses showed any significant association between RET/PTC expression and the clinical and pathological features of the cancers. CONCLUSION: While this prevalence of the RET/PTC is less than RET/PTC frequency seen after Chernobyl in Belarus, its prevalence in our region is also high (66.3%). As a result, no significant correlation was found in between prognosis and RET/PTC frequency. PMID- 18393129 TI - Oxidative stress markers in young patients with polycystic ovary syndrome, the relationship between insulin resistances. AB - OBJECTIVE: Polycystic ovary syndrome is a syndrome of ovarian dysfunction. Oxidative stress, inflammation and endothelial cell activation are thought to play concomitant roles in the pathogenesis of the above diseases particularly in the development of atherosclerotic lesions. RESEARCH DESIGN AND METHODS: We studied 58 polycystic ovary syndrome patients and age-matched 25 healthy controls consisting of women that have regular, ovulatory cycles and normal androgen levels. Homeostasis Model Assessment-Insulin Resistance for this study was taken as 1.75 that is the upper level of confidence interval of %95 of the mean of the healthy group. PCOS patients were divided into two groups as for below the cut off level (<1.75) and above the cut-off level (> or =1.75). hs-CRP, fibrinogen, malondialdehyde, nitric oxide and disulfide level results were compared both in PCOS and control groups. RESULTS: In this study, sensitive CRP was found to be statical significantly higher in polycystic ovary syndrome groups whose Homeostasis Model Assessment-Insulin Resistance were > or =1.75 and <1.75 when compared to the control group. But, no significantly correlation was determined between malondialdehyde, nitric oxide and disulfide levels and CRP elevation. CONCLUSIONS: In our study, because those participants were young and non- obese patients with PCOS, malondialdehyde, nitric oxide and disulfide levels and Carotid Artery Intima-Media Thickness measurements as a pre-indicator of cardiovascular disease were not found to be different from those of the controls. PMID- 18393130 TI - Genetic polymorphisms of beta2- and beta3-adrenergic receptor genes associated with characteristics of the metabolic syndrome in black South African women. AB - BACKGROUND: Genetic variation in the beta2 (ADRB2) and beta3 (ADRB3) adrenergic receptor genes are associated with obesity and insulin resistance. To further elucidate the role of these genes in the pathophysiology of obesity the present study investigated associations between certain polymorphisms in ADRB2 and ADRB3 and parameters of carbohydrate and lipid metabolism in a population of African origin. MATERIAL AND METHODS: Data of 102 black South African women obtained in the POWIRS (Profile of Obese Women with the Insulin Resistance Syndrome) study were used. Endpoint measurements included several anthropometric variables, resting blood pressure, plasma glucose, insulin, free fatty acids (FFA), ghrelin, leptin and lipids, and insulin resistance as estimated by the homeostasis model assessment (HOMA-IR) index. Polymorphisms were analyzed via PCR based methods. RESULTS: The percentage body fat was significantly lower (p< or =0.05) and the FFA significantly higher (p< or =0.05) in lean subjects (BMI< or =25 kg/m2) with the Glu27 variant allele compared to subjects with the Gln27 wildtype allele of the ADRB2 gene. In contrast, the variant allele of the ADRB2 gene was significantly positive associated (p< or =0.05) with the HOMA-IR-index in overweight black African women (BMI>25 kg/m2). No significant differences in parameters of the metabolic syndrome were apparent between subjects with the wildtype and variant alleles in the ADRB3 gene. CONCLUSION: The presence of the Glu27 and Arg64 polymorphisms of the ADRB2 and ADRB3 genes are not directly related to indices of the metabolic syndrome. PMID- 18393131 TI - The role of immunohistochemistry in histopathological diagnostics of clinically "silent" incidentally detected adrenal masses. AB - BACKGROUND: The detectability of adrenal incidentalomas (incidentally found adrenal tumours) in the whole population is estimated at 0.1%; 0.42% in non endocrine patients and at 4.3% in oncologically diagnosed ones. Even up to 16% of incidentalomas of adrenal glands can be malignant lesions. The issue of crucial importance is the histopathological differentiation between benign lesions and malignant tumours of the adrenal cortex and medulla. OBJECTIVES: To evaluate whether the immunohistochemical analysis of the expression of p53, p21, PCNA and Ki67 in the tumour's tissue can be useful in the histopathological diagnostics of adrenal incidentalomas and whether it is important for prognosis. MATERIAL AND METHODS: Our series consisted of 74 tumour samples from 164 patients operated for incidentalomas. There were 43 cortical adenomas, 11 cortical adrenocarcinomas and 20 PHEOs (including 5 malignant lesions). Using monoclonal antibodies, the expression of p53, p21, PCNA and Ki67 was evaluated. RESULTS: We found a statistically significant correlation between the expression of p53, p21, Ki67 and the differential diagnosis of adrenal cortical adenoma and adrenocortical carcinoma (for proteins: p53 p=0.010, for p21 p=0.010, for Ki67 p<0.001). The statistical significant correlation between PCNA protein and diagnosis of adrenal cortical adenoma and adrenocortical carcinoma was not found. The statistically significant correlation between p21, PCNA proteins and the diagnosis of benign and malignant PHEOs was not estimated. There was no expression of Ki67 or p53 protein above the assumed level in benign and malignant pheochromocytomas. The statistically significant correlation between p53, p21, PCNA or Ki67 and the occurrence of metastases in adrenocarcinoma and malignant PHEOs was not found. PMID- 18393132 TI - [Relatives as a means of stroke rehabilitation]. PMID- 18393133 TI - [Side effects of psychotherapeutic intervention for caregivers of stroke patients: do patients also benefit?]. AB - AIM: The present study aimed at investigating whether stroke patients benefit from their spouses attending a psychotherapeutic group intervention, in terms of improved functioning level and quality of life. METHOD: Within the scope of a panel study with control group design, n = 124, stroke patients were interviewed using standardized questionnaires, regarding competences of daily life (PCRS) and quality of life (WHOQoL). In the intervention group, the patients' spouses attended a psychotherapeutic group intervention; the spouses of two control groups merely received illness-related information. Data was evaluated statistically by means of multiple regression analysis. RESULTS: No statistically significant effects of the caregiver intervention on the patients were found. However, patients of the intervention group tended to report improved quality of life in the course of the study period, whereas subjective quality of life worsened for patients of the control groups. In all groups, impairments of everyday-life competences increased. DISCUSSION: The results of this study suggest that psychotherapeutic group intervention for caregivers also has positive side effects on the patients. Further research is necessary in order to provide a better understanding of the mechanisms at work behind these positive side effects. Assistance concepts for spouses of stroke patients should utilize such intermediary effects more systematically. PMID- 18393134 TI - [The Basel Screening Instrument for Psychosis (BSIP): development, structure, reliability and validity]. AB - BACKGROUND: Early detection of psychosis is of growing clinical importance. So far there is, however, no screening instrument for detecting individuals with beginning psychosis in the atypical early stages of the disease with sufficient validity. We have therefore developed the Basel Screening Instrument for Psychosis (BSIP) and tested its feasibility, interrater-reliability and validity. AIM: Aim of this paper is to describe the development and structure of the instrument, as well as to report the results of the studies on reliability and validity. METHOD: The instrument was developed based on a comprehensive search of literature on the most important risk factors and early signs of schizophrenic psychoses. The interraterreliability study was conducted on 24 psychiatric cases. Validity was tested based on 206 individuals referred to our early detection clinic from 3/1/2000 until 2/28/2003. RESULTS: We identified seven categories of relevance for early detection of psychosis and used them to construct a semistructured interview. Interrater-reliability for high risk individuals was high (Kappa .87). Predictive validity was comparable to other, more comprehensive instruments: 16 (32 %) of 50 individuals classified as being at risk for psychosis by the BSIP have in fact developed frank psychosis within an follow-up period of two to five years. CONCLUSIONS: The BSIP is the first screening instrument for the early detection of psychosis which has been validated based on transition to psychosis. The BSIP is easy to use by experienced psychiatrists and has a very good interrater-reliability and predictive validity. PMID- 18393135 TI - [Progress of social integration of mentally disabled persons in Japan]. AB - The social situation of the mentally disabled in Japan was characterized by living in closed sanatoriums with only few possibilities to participate in normal social life until the 80 (ths) of the last century. However, important improvements regarding the social integration have been achieved over the last years. Revisions of existing laws, private initiatives like working places for the disabled and other community associated facilities have been of special importance and have led to a better integration of the disabled in the normal social life. In the report examples of successful initiatives are described. The paid labour in the working facilities seems to be of high importance as they permit an economic and social autonomy and a higher self-respect. PMID- 18393138 TI - Hot topics II: an editorial collection of current issues and controversies in thrombosis and hemostasis. PMID- 18393136 TI - [Quality management in psychiatry]. PMID- 18393139 TI - Measuring the quality of journals and journal articles: the impact factor tells but a portion of the story. AB - Much emphasis has been placed on the IMPACT FACTOR, a measure of journal article citation rates used as a surrogate marker of both journal and article quality. There is little doubt that the impact factor is an important audit of journal article usage, as it in essence provides a measure of the level of peer attention being given to articles within journals and (by extrapolation) of the level of attention being given to the journal containing those articles. However, the impact factor has its limitations and only tells a very small fraction of the overall story regarding the utility of journals and the articles within them. In addition, the impact factor can be easily manipulated. The current article includes a brief review of the current and past uses and abuses of the impact factor and describes some of its strengths and limitations. In addition, a review of past publications, primarily from this journal, has been undertaken to help show the potential use of alternative measures of journal utility, such as Internet-based journal sessions and article downloads. The evaluation of previously published articles also helps serve to illustrate, by example, some of the limitations to the use of the impact factor as the sole determinant of a journal's "quality." PMID- 18393140 TI - Metabolic modulation of inflammation-induced activation of coagulation. AB - The bidirectional interaction between coagulation and inflammation, which is relevant in various disease states that are dominated by systemic inflammatory responses, such as severe infection or chronic vascular disease, is modulated by metabolic factors. Changes in lipoprotein metabolism affect the inflammation induced activation of coagulation, which may have impact on downstream effects, including organ dysfunction and survival. Likewise, glucose and insulin seem to influence inflammation-induced effects on coagulation and fibrinolysis. Hyperglycemia affects inflammation-induced and tissue factor-driven activation of coagulation, whereas hyperinsulinemia modulates the fibrinolytic response. There are indications that this modulatory role of metabolic factors in inflammation and coagulation may also have an impact on clinical outcome in various disease states. PMID- 18393141 TI - Are fibrin-related markers useful for the diagnosis of thrombosis? AB - Fibrin-related markers (FRMs), such as fibrin and fibrinogen degradation products (FDPs), D-dimer, and soluble fibrin (SF), are considered to be useful for the diagnosis of thrombosis. However, the evidence for the making of a diagnosis of thrombosis based on FRMs is, as yet, not fully established. Levels of FRMs are significantly elevated in patients with thrombosis, such as deep vein thrombosis, disseminated intravascular coagulation, and so on. In Japan, the D-dimer assay test result might be 2-fold higher than results for those assays commonly used in Europe and North America. The levels of SF are significantly elevated in patients before the onset of thrombosis, thus suggesting that the SF assay is useful not only for the diagnosis of thrombosis but also for diagnosing a prethrombotic state. Overall, elevated levels of FRMs indicate a high risk for thrombosis, and they are thus considered to be useful for the diagnosis of thrombosis. PMID- 18393142 TI - Current and future prospects for anticoagulant therapy: inhibitors of factor Xa and factor IIa. AB - Indirect systemic and direct oral factor Xa and direct oral factor IIa inhibitors with improved pharmacologic profiles compared with heparins and vitamin K antagonists are currently in clinical development. This overview focuses on the indirect antithrombin dependent pentasaccharide derivatives of idraparinux and on the most advanced oral direct inhibitors to factor Xa (rivaroxaban and apixaban) and IIa (dabigatran). Specifically, the results of dose-finding studies for the prevention of venous thromboembolism after elective orthopedic surgery, the results of dose-finding studies for treatment of acute venous thromboembolism including prolonged prophylaxis of recurrent events, and the designs of ongoing clinical trials are reviewed. PMID- 18393143 TI - Survival of heparins, oral anticoagulants, and aspirin after the year 2010. AB - The conventional management of thrombotic and cardiovascular disorders is based on the use of heparin, oral anticoagulants, and aspirin. Despite remarkable progress in life sciences, these drugs still remain a challenge and a mystery to us, and their use is far from optimized. The development of low-molecular-weight heparins and the synthesis of heparinomimetics, such as the chemically synthesized pentasaccharide, represent a refined use of heparin. Additional drugs from this knowledge will continue to develop; however, none of these drugs will ever match the polypharmacology of heparin. Aspirin still remains the leading drug in the management of thrombotic and cardiovascular disorders. The newer antiplatelet drugs such as adenosine diphosphate receptor inhibitors, glycoprotein IIb/IIIa inhibitors, and other specific inhibitors have limited effects and have been tested in patients who have already been treated with aspirin. Warfarin provides a convenient and affordable approach in the long-term outpatient management of thrombotic disorders. The optimized use of these drugs still remains as the approach of choice to manage thrombotic disorders. The new anticoagulant targets, including specific sites in the hemostatic network such as tissue factor, individual clotting factors (IIa, VIIa, IXa, Xa, XIIa, and XIIIa), recombinant forms of serpins (antithrombin, heparin cofactor II, and tissue factor pathway inhibitors), recombinant activated protein C, thrombomodulin, and site-specific serine protease inhibitor complexes have also been developed. There is a major thrust on the development of orally bioavailable anticoagulant drugs (anti-Xa and anti-IIa agents), which are slated to replace oral anticoagulants. Both the anti-factor Xa and antithrombin agents have been developed for oral use and have provided impressive clinical outcomes in sponsor trials for the postsurgical prophylaxis of venous thrombosis; however, safety concerns related to liver enzyme elevations and thrombosis rebound have been reported with their use. For these reasons, the U.S. Food and Drug Administration did not approve the orally active antithrombin agent ximelagatran for several indications. The synthetic pentasaccharide (fondaparinux) has undergone an aggressive clinical development. Unexpectedly, fondaparinux also produced major bleeding problems at minimal dosages. Fondaparinux represents only one of the multiple pharmacologic effects of heparins. Thus, its therapeutic index will be proportionately narrower. The newer antiplatelet drugs have added a new dimension in the management of thrombotic disorders. The favorable clinical outcomes with aspirin and clopidogrel have validated cyclooxygenase (COX)-1 and P2Y (12) receptors as targets for new drug development. Prasugrel, a novel thienopyridine, cangrelor, and AZD 6140 represent newer P2Y (12) antagonists. Cangrelor and AZD 6140 are direct inhibitors, whereas prasugrel requires metabolic activation. Though clinically effective, recent results have prompted a closure of a large clinical trial with prasugrel due to bleeding. The newer anticoagulant and antiplatelet drugs are attractive for several reasons; however, none of these are expected to replace the conventional drugs in polytherapeutic approaches. Heparins, warfarin, and aspirin will continue to play a major role in the management of thrombotic and cardiovascular disorders beyond 2010. PMID- 18393144 TI - Differentiating low-molecular-weight heparins based on chemical, biological, and pharmacologic properties: implications for the development of generic versions of low-molecular-weight heparins. AB - Low-molecular-weight heparins (LMWHs) are polypharmacologic drugs used to treat thrombotic and cardiovascular disorders. These drugs are manufactured using different chemical and enzymatic methods, resulting in products with distinct chemical and pharmacologic profiles. Generic LMWHs have been introduced in Asia and South America, and several generic suppliers are seeking regulatory approval in the United States and the European Union. For simple small-molecule drugs, generic drugs have the same chemical structure, potency, and bioavailability as the innovator drug. Applying this definition to complex biological products such as the LMWHs has proved difficult. One major issue is defining appropriate criteria to demonstrate bioequivalence; pharmacopoeial specifications alone appear to be inadequate. Whereas available generic versions of LMWHs exhibit similar molecular and pharmacopoeial profiles, marked differences in their biological and pharmacologic behavior have been noted. Preliminary studies have demonstrated differences in terms of anti-Xa activity and tissue factor pathway inhibitor release after subcutaneous administration, as well as antiplatelet and profibrinolytic effects. The current data emphasize the need to consider multiple functional parameters when defining bioequivalence of biologic drugs with complex structures and activities and also underscore the importance of further pharmacologic studies involving animal models and human clinical trials. The U.S. Food and Drug Administration and the European Medicine Evaluation Agency are currently developing guidelines for the acceptance of biosimilar agents including LMWHs. Until such guidelines are complete, generic interchange may not be feasible. PMID- 18393145 TI - The laboratory diagnosis and clinical management of patients with heparin-induced thrombocytopenia: an update. AB - Heparin-induced thrombocytopenia (HIT) is a serious adverse effect of heparin exposure that can progress to severe thrombosis, amputation, or death. HIT is an immune response in which antibodies cause platelet activation, platelet aggregation, the generation of procoagulant platelet microparticles, and activation of leukocytes and endothelial cells. Early diagnosis based on a comprehensive interpretation of clinical and laboratory information is important to improve clinical outcomes. However, limitations of the laboratory assays and atypical clinical presentations can make the diagnosis difficult. Clinical management of patients with HIT is with a non-heparin anticoagulant such as a direct thrombin inhibitor or danaparoid followed by a vitamin K antagonist for long-term treatment. The new anti-factor Xa drugs (fondaparinux, rivaroxaban, apixaban) and other non-heparin antithrombotic agents can potentially be used for the treatment of HIT if clinically validated. Important drug-specific limitations and dosing and monitoring guidelines must be respected for patient safety. Issues still exist regarding the optimal clinical management of HIT. PMID- 18393146 TI - Can we predict bleeding? AB - The prior identification of subjects who are likely to bleed excessively when subjected to operative surgery and other invasive procedures is desirable. Frequently, reliance is placed on laboratory-based screening tests of blood coagulation for this purpose. However, published evidence does not support this approach as the tests are not fit for purpose, and their sensitivity and specificity are low. Some more global assays may have use in the diagnostic workup in subjects with hemorrhage, but none has been established to date as an efficient method for prediction of bleeding in unselected populations. There is renewed interest in the use of the clinical history for the prediction of bleeding. Recent reports suggest that when a structured questionnaire is employed to derive a bleeding score, the positive predictive value of the approach for the detection of bleeding disorders is high. PMID- 18393147 TI - A critical review on the use of recombinant factor VIIa in life-threatening obstetric postpartum hemorrhage. AB - The objective of this review was to evaluate and summarize the current literature on the unlicensed use of the novel agent recombinant activated factor VII (rFVIIa) in the management of major postpartum hemorrhage. After a systematic electronic search without temporal limits on MEDLINE, EMBASE, OVID and SCOPUS, the bibliographic references of all retrieved studies and reviews were additionally assessed for further reports of clinical trials. Unpublished works were also identified by searching abstracts from the most eminent conferences on this topic. In total, there were 31 studies that fulfilled our inclusion criteria. These studies incorporated 118 cases of massive postpartum hemorrhage treated with rFVIIa. The median age of the patients was 31.4 years, and cesarean section appeared to increase the risk of postpartum hemorrhage. At a median dose of 71.6 mug/kg, rFVIIa was reported to be effective in stopping or reducing bleeding in nearly 90% of the reported cases. Based on the evidence from the literature, we give some recommendations on the use of rFVIIa in massive postpartum hemorrhage. Nevertheless, although these reports suggest the potential role of rFVIIa in treating massive postpartum hemorrhage refractory to standard therapy, we advise particular caution in interpreting these results, as they are derived from few and uncontrolled studies. Further evidence is needed using well designed clinical trials to better assess the optimal dose, the effectiveness, and the safety of rFVIIa in such critical bleeding conditions. PMID- 18393148 TI - Phenotypic identification of platelet-type von Willebrand disease and its discrimination from type 2B von Willebrand disease: a question of 2B or not 2B? A story of nonidentical twins? Or two sides of a multidenominational or multifaceted primary-hemostasis coin? AB - Platelet-type von Willebrand disease (PT-VWD) and type 2B von Willebrand disease (2B-VWD) have different etiologies although both present with a similar clinical bleeding and basic laboratory phenotype. Both PT-VWD and 2B-VWD represent gain-of function mutations that lead to enhanced binding between plasma von Willebrand factor (VWF) and its platelet ligand, glycoprotein Ib alpha (GP1BA). However, 2B VWD results from a functionally abnormal VWF molecule arising from mutations in the VWF gene, whereas PT-VWD is caused by hyperresponsive platelets resulting from mutations in the platelet GP1BA gene. A definitive diagnosis of PT-VWD versus 2B-VWD is critical for treatment decisions (as differential therapies might be respectively required) and also for family counseling. However, laboratory discrimination of PT-VWD versus 2B-VWD is problematic because simple phenotypic testing will not permit their differentiation, and the more complex testing approaches that might permit their differentiation are rarely applied, or are perhaps poorly applied. Although differential identification of PT-VWD versus 2B-VWD can most definitively be achieved by identifying the gene defect at either the VWF or GP1BA loci, such tests are not commonly available, not always successful, and even if available and successful might not be readily available to serve time-critical treatment decisions. Accordingly, simple laboratory tools to enable discrimination of the two disorders would be a valuable addition to the test repertoire of the hemostasis laboratory. This article provides a review of PT-VWD-related literature and an overview of a phenotypic laboratory test process that should enable the effective identification of PT-VWD and its discrimination from 2B-VWD. PMID- 18393149 TI - The phenotypic heterogeneity of severe hemophilia. AB - It has been long recognized that 10 to 15% of patients with "phenotypically characterized" severe hemophilia (< 1% clotting factor activity) have relatively mild disease clinically. Not all these patients have frequent spontaneous bleeding, and even among those who bleed, the extent of joint damage tends to vary considerably. The basis for this difference has not been completely understood. This article reviews the literature on possible determinants of phenotypic variation in patients with severe hemophilia. Apart from the well recognized associations of the level of residual clotting factor activity, pharmacokinetics of administered clotting factor concentrates, and presence of prothrombotic markers, there is evidence to suggest that variations in other coagulation proteins as assessed in tests of global hemostasis as well as the fibrinolytic system can affect the clinical severity of bleeding. We also hypothesize that mediators of the inflammatory response in the synovium are likely to impact the severity of joint damage in these patients. One of the major issues in the management of hemophilia today is to decide on ways in which therapy, particularly the initiation and intensity of prophylaxis, can be individualized. A detailed understanding of all factors that may contribute to joint damage in severe hemophilia could help us in tailoring therapy for these individuals. PMID- 18393150 TI - A facilitated method for endoscopic interventions at the bile duct after Roux-en Y reconstruction using double balloon enteroscopy. AB - Endoscopic retrograde cholangiography (ERC) after Roux-en-Y reconstruction and modified BII surgery or duodenopancreatectomy is considerably more difficult than ERC under normal anatomic conditions. If the common bile in the afferent loop cannot be reached by a common lateral-viewing duodenoscope because of excessive intestinal length, it has recently become possible to use double balloon enteroscopy (DBE) for ERC to reach the common bile duct. Cannulating the bile duct via DBE in these postoperative settings remains one of the most difficult ERCP manipulations because of the lack of an Albarran lever and the use of extra long ERCP accessories. Here, we report on a facilitated method for endoscopic interventions at the bile duct in postoperative settings with a long afferent loop using DBE. For facilitation of interventions the enteroscope can be exchanged for a 110-cm-long conventional gastroscope after incision of the overtube in three quarters of its circumference. Care has to be taken that the pressure line for the balloon remains intact. The huge benefit of this facilitated method is the use of standard endoscopic material like guides, catheters and papillotomes. PMID- 18393151 TI - [MR-colonography in hospitalized patients: feasibility and sensitivity]. AB - INTRODUCTION: Besides conventional colonoscopy, CT and MR colonography offer alternate virtual imaging modalities of the colon. The sensitivity of CT colonography, which is associated with radiation exposure, has been published in prior, large studies. Regarding MR colonography, in particular dark lumen MR colonography with the rectal administration of a water enema as a contrast agent, only limited published data exist. The goal of this study was to compare MR colonography with conventional colonoscopy in the detection of colorectal polyps. In addition the feasibility and image quality in unselected hospitalised patients were assessed. PATIENTS/METHODS: Included were 103 hospitalised patients who had to undergo colonoscopy for various indications. Immediately prior to conventional colonoscopy, MR colonography with rectal water enema and additional intravenous administration of contrast material was performed. Detection rates for polyps and adenomas were documented with both imaging modalities. Image quality and completion rates (practicability) and other (incidental) findings were also recorded. RESULTS: In 15 of 103 patients the MR examination could not be done or was only partially completed. The detection rate of MR colonography for polyps (adenomas) was 2% (4%) for polyps (adenomas) up to 5 mm in diameter, 38% (56%) for polyps (adenomas) 6-10 mm in diameter and 89% (89%) for polyps (adenomas) up to 11 mm in diameter. One flat carcinoma seen with conventional coloscopy was missed on MR colonography. CONCLUSIONS: MR colonography offers the possibility of imaging the colon without exposure to radiation. Polyps and adenomas are detected, similar to the detection rate of CT colonography, with adequate sensitivity only if they are larger than 10 mm in diameter. Therefore this imaging technique is not (yet) suitable as a screening test. Additional limitations are the necessary cooperation of the patient which can reduce the practicability and image quality in selected patients. Further studies like the just started German multicentre trial are needed to assess the position of MR colonography. PMID- 18393152 TI - Acute abdominal pain caused by spontaneous hemorrhagic infarction of a solitary plexiform neurofibroma of lesser omentum. AB - A 32-year-old female patient visited the emergency room because of sudden onset of abdominal pain, which was found to be induced by hemorrhagic infarction of a plexiform neurofibroma arising from the lesser omentum. No other classic clinical characteristics of systemic von Recklinghausen's disease had been seen up until now. The tumor, classified as sporadic plexiform neurofibroma, rarely occurs in the digestive tract. This diagnosis was confirmed by exploratory laparotomy with complete resection of the tumor. Here, we describe the rare case of a sporadic intra-abdominal plexiform neurofibroma and discuss the clinical characteristics and management of plexiform neurofibroma. PMID- 18393153 TI - Rectal prolapse as initial clinical manifestation of colon cancer. AB - Rectal prolapse as the initial clinical manifestation of colorectal cancer is uncommon. We describe the case of a 75-year-old woman who was diagnosed as having adenocarcinoma of the sigmoid colon after presenting with complete rectal prolapse. The tumor caused rectosigmoid intussusception and then it prolapsed out through the anus. She underwent rectosigmoidectomy and rectopexy. The postoperative course was uneventful. The relationship between colorectal cancer and rectal prolapse has not been clearly established. This case report describes an unusual presentation of colorectal cancer. It suggests that rectal prolapse can present as the initial symptom of colorectal cancer and may also be a presenting feature of the occult intra-abdominal pathology. The importance of adequate investigation such as colonoscopy should be emphasized in patients who develop a new onset of rectal prolapse. PMID- 18393154 TI - [Diagnosis of an accessory liver lobe established by magnetic resonance imaging guided core biopsy]. AB - An accessory liver is a rare developmental anomaly. There are two subtypes: the accessory liver lobe (ALL), which is connected to the liver and the true ectopic liver. An accessory liver is usually of no clinical relevance in spite of the very rare cases in which torsion or other complications may cause serious problems. This anomaly should not be misinterpreted as a malignant tumour. We report the case of an accessory liver lobe, located below the left diaphragm, which was misdiagnosed as a metastasis of a known malignant melanoma. The diagnosis ALL was established by a biopsy under guidance of magnetic resonance tomography (MRT). In our opinion, an accessory liver and its appearance should be known, to include this entity in differential diagnosis of perihepatic masses. PMID- 18393155 TI - [Current status of ultrasound in gastroenterology--bowel and upper gastrointestinal tract--part 2]. AB - Ultrasound has gained acceptance in the diagnosis of diseases of the gastrointestinal tract beside the classical methods such as endoscopy and X-ray. In a previous publication we discussed the use of ultrasound in emergency diagnostics (e. g., acute appendicitis, diverticulitis/peridiverticulitis, ileus, invagination and perforation) (part 1). Because of the vast extent of this topic, in this overview we will focus on the current role of ultrasound in the detection and assessment of chronic inflammatory bowel diseases, rare forms of colitis (e. g., bacterial, pseudomembranous and neutropenic colitis as well as intestinal tuberculosis), ischaemic bowel diseases as well as diseases of the upper gastrointestinal tract. In chronic inflammatory bowel diseases, ultrasound can give important additional information such as extension, activity, complication (fistula, abscess, stenosis) and in differential diagnosis. It plays an important role in follow-up investigations and can possibly reduce the number of endoscopic examinations. There is still some debate going on about the significance of colour Doppler ultrasound in assessing the activity and differentiation of stenosis. Furthermore, ultrasound is used as a method to guide interventional therapies for abscesses (puncture and drainage). Colour Doppler ultrasound can diagnose ischaemic bowel diseases and also differentiate these from other aetiologies. Ultrasound plays a greater role in the follow-up and assessment of chronic intestinal ischaemia. In the diagnosis of stomach diseases under favourable conditions ultrasound can show changes of the stomach wall, tumours, ulcers and their complication (perforation, penetration) and disturbances of the motility. But an exclusion is not possible. PMID- 18393156 TI - [[18F]-FDG-PET in the diagnostics of gastrointestinal tumors]. AB - Positron emission Tomography (PET) with 2-[(18)F]-fluoro-2-deoxy-d-glucose (FDG) is a functional imaging technique with increasing value in special diagnostic fields of gastrointestinal tumours. In the initial staging of esophageal and gastric cancer, FDG-PET is useful in the staging of patients with advanced but local resectable disease. The detection of distant metastases results in an up staging, and these patients should not be treated by surgery. Furthermore, FDG PET is sufficient for monitoring early therapy responses after neoadjuvant treatment and enables one to select non-responders who may benefit from therapy alterations. Major indications for FDG-PET in patients with rectal carcinoma are therapy monitoring and diagnosis of relapses, especially the differentiation between tumour and scar and also the localisation of tumour manifestations in cases with increasing tumour markers. FDG-PET is very efficient in the imaging of pulmonal and hepatic metastases of colorectal cancer but not in lymph node staging. In diagnostic procedures for pancreatic carcinoma, FDG-PET can be recommended to explore the dignity of pancreatic lesions and in the imaging of tumour relapses. For gastrointestinal stroma tumours, FDG-PET is useful for the monitoring of therapy and the initial staging. For imaging of hepatocellular carcinoma and carcinoma of the gall bladder, FDG-PET is not sufficient. PMID- 18393158 TI - Functional magnetic resonance imaging and cortical mapping in motor cortex tumor surgery: complementary methods. AB - Functional magnetic resonance imaging (fMRI) and direct electrocortical stimulation (DES) are the most commonly used means of analyzing the functional brain topography prior to surgery in the vicinity of Brodmann area 4. No consensus has been established in the literature about the significance of both procedures in reducing operative morbidity. The study presented here was conducted in 30 patients with tumors in the area of the primary motor cortex. Blood oxygen level dependent (BOLD) sequences were preoperatively established with a standardized paradigm. Intraoperatively motor mapping was performed with DES. The results of both methods were digitally matched with a frameless image guidance system. Correlations between the results of fMRI and of DES were analyzed. Furthermore, the potential influences of the size, position, and histology of the lesions on the mapping results were analyzed and the motor outcome was evaluated. The mean deviation between the results of fMRI and of DES was 13.8 mm (range: 7-28 mm). This deviation was independent of the histology, size, or location of the corresponding lesion. The individual variability of the analysis threshold value for the evaluation of the BOLD sequences led to a considerable topographical inaccuracy. As complementary methods, fMRI contributes to estimating the operational risk, while DES is performed when the results of MRI and fMRI suggest an immediate proximity of the tumor to motor areas. PMID- 18393159 TI - Preoperative predictors for the return to work of herniated disc patients. AB - This study assessed socio-demographic, physical, psychological, and work-related factors predicting the ability to work 6 months after operation in disc herniation patients. After nucleotomy 214 patients answered questionnaires on job satisfaction, their desire for a disability pension, preoperative sick leave, and completed the QLQ-C30, and SCL-27A questionnaires. Additionally, data on the severity of injury and duration of pain were extracted from the clinical reports. 182 patients answered the follow-up questionnaire 6 months after operation by telephone. Socio-demgraphic and psychological factors did not influence the return to work. Factors found to be related to the ability to work were job satisfaction, preoperative sick leave, pain, and the desire for a disability pension. Using multiple regression analysis, the ability to work 6 months after rehabilitation was predicted on the basis of job satisfaction, a preoperative sick leave of less than 6 weeks, and a low pain intensity preoperatively. The results are discussed with respect to their socioeconomic implications. PMID- 18393160 TI - Radiosurgery/stereotactic radiotherapy in the therapeutical concept for skull base meningiomas. AB - OBJECTIVE: Microsurgical resection is still the treatment of choice for skull base meningiomas. But the risk of postoperative neurological deficits is high, and in many of these cases complete tumor removal cannot be achieved. Therefore recurrences are even more probable. Stereotactically guided radiation therapy - radiosurgery (RS) or stereotactic radiotherapy (SRT) - offers an additional or alternate treatment option for those patients. We evaluated local control rates, symptomatology, and toxicity. PATIENTS AND METHODS: 224 patients were treated with stereotactically guided radiation techniques in two departments between 1997 and 2003. 129 of 224 had recurrences after 1 to 3 prior tumor resections and 95 of 224 were treated with SRT/RS alone. 87.9% of cases had benign, 7.8% had atypical and 4.3% had malignant meningiomas. RS was only applied in 11 cases. Tumor volumes ranged from 0.16 ccm to 3.56 ccm. The other 213 patients had larger tumor volumes of up to 135 ccm or a meningioma close to optical structures. Therefore 183 cases were treated with SRT in normal fractions of 1.8-2 Gy in single doses up to 60 Gy. Hypofractionated SRT with single fraction doses of 5 or 4 Gy was applied in 30 cases. Follow-up data were available in 181 skull base meningiomas and the progression-free and overall survival rates, the toxicity and symptomatology were evaluated. RESULTS: The median follow-up was 36 months. The overall survival and the progression-free survival rates for 5 years were 92.9%, and 96.9%, respectively. Two tumor progressions have occurred to date but further follow up is required. Tumor volumes (TV) had shrunk about by 19.7% at 6 months (p<0.0001) and by 23.2% at 12 months (p<0.01) after SRT/RS. In 95.6% the symptoms had improved or were stable. Clinically significant acute toxicity (grade III) was seen in only 1 case (2.7%). Some patients developed late toxicity: 8.8% had grade I, 4.4% had grade II and 1.1% had grade III. No other neurological deficits occurred during follow-up. CONCLUSION: SRT and RS offer an additional or alternative treatment option with a high efficacy and few side effects for the tumor control of skull base meningiomas. An individual and interdisciplinary decision respecting treatment is needed for each patient. In cases of large TV (>4 ccm), tumors adjacent to critical structures (<2 mm) or in high-risk patients the use of SRT offers greater benefits. PMID- 18393161 TI - Extensive brain swelling with neurological deterioration after intracranial meningioma surgery - venous complication or 'unspecific' increase in tissue permeability. AB - OBJECTIVE: We retrospectively analyzed a series of patients who deteriorated after resection of an intracranial meningioma due to extensive brain edema (EBE) with regard to etiology of the neurological worsening and outcome. METHODS: Out of a series of 376 consecutive patients who underwent resection of an intracranial meningioma, 13 (3.5%) experienced postoperative deterioration due to EBE which necessitated prolonged artificial ventilation, tracheal reintubation, or decompressive craniectomy. Clinical data, radiological findings, operative records and follow-up data of these patients were retrospectively reviewed. RESULTS: The study revealed two different patient groups: Patients in group A (n=7) demonstrated edema due to typical venous infarction (VI). A decompressive craniotomy was performed in all but one patient in this group; nonetheless, an acceptable neurological outcome (Glasgow Outcome Scale (GOS) 4) was achieved in only two cases. Patients in group B (n=6) deteriorated due to an EBE of unknown etiology. Mean tumor volume in this group was higher when compared to group A (75 ml vs. 30 ml). In addition, 83% of patients in group B displayed extensive preoperative peritumoral edema compared to only 14% in group A. Three patients in group B required decompressive surgery; however, neurological outcome was more favorable in this group as 83% achieved a GOS of 4 or 5. CONCLUSION: Extensive brain swelling during or after intracranial meningioma surgery may be due to VI or possibly due to increased postoperative tissue permeability. It is recommended to preserve all venous structures as patients with VI had an unfavorable neurological outcome. PMID- 18393163 TI - Clinical results and surgical technique for the treatment of extreme lateral lumbar disc herniations: the minimally invasive microscopically assisted percutaneous approach. AB - OBJECTIVE: Operative techniques for the treatment of extreme lateral lumbar disc herniations (ELLDH) have ranged from an interlaminar approach with subtotal or total facetectomy to an enlarged midline approach and various paraspinal lateral, as well as endoscopic approaches. In contrast to purely endoscopic techniques, use of a working tube together with an operative microscope ("minimally invasive microscopically assisted percutaneous approach") combines the advantages of three dimensional visual control with the minimal surgical trauma of an endoscopic approach. We present this technique and the clinical results in patients with extreme lateral lumbar disc herniation (ELLDH). METHODS: In a retrospective case series we analyzed the data of 73 consecutive patients with a disc herniation within or lateral to the intervertebral foramen. All of them were operated on using the surgical microscope after blunt dilatation of the paraspinal muscles and insertion of a tubular retractor system (METRx, Medtronic Sofamor Danek). Fifty-nine patients were included in the follow-up study. Follow-up ranged from 10 to 47 months (mean 30.4) and was evaluated after telephone inquiry by the co author using the modified MacNab criteria . RESULTS: The results with respect to improvement of preoperative symptoms were excellent in 23 patients (39%), good in 25 patients (42.4%), fair in 8 patients (13.5%) and poor in 3 patients (5.1%). Twenty-four patients had still been working prior to surgery, and 20 returned to work after 4-8 weeks. CONCLUSION: The paraspinal approach for extreme lateral lumbar disc herniations using the minimally invasive microscopically assisted percutaneous technique is reasonable and safe and thus a good alternative to open surgical procedures. PMID- 18393162 TI - The value of intraoperative three dimensional fluoroscopy in anterior decompressive surgery of the cervical spine. AB - PURPOSE: Intraoperative use of the mobile Siremobil Iso-C3D C-arm (Siemens AG, Medical Solutions, Erlangen) considerably improves the information available during cervical spine surgery. We report our experiences with the Iso-C3D fluoroscopic unit during anterior decompressive surgery of the cervical spine. METHODS: We used the mobile Siremobil Iso-C3D C-arm during decompressive cervical spine surgery. The study included 25 patients (22 males, 3 females) with degenerative cervical stenosis. Mean age was 55.9 years (42-73 years). Eighteen patients were surgically treated for one-level, six for two-level and one for three-level disease. Intraoperative 3D imaging was performed to evaluate the extent of bony decompression and to assist correct positioning of the cages when the surgeon believed that sufficient decompression had been achieved. RESULTS: Visualization of the extent of bone removal was good in all patients. In 3 patients, insufficient bony decompression with persisting dorsal osteophytic spurs was noticed on sagittal and axial images. In these patients, surgery had to be continued. Successful decompression was proved thereafter by a second scan. The quality of the images of the cervical spine was sufficient, although slightly inferior to that of a CT scan. CONCLUSION: The Siremobil Iso-C3D provides intraoperative 3D images of bony structures of the cervical spine. Although the imagine quality is inferior to that of a CT, in our series surgical revisions could be avoided in 12.5% of the patients on the basis of these intraoperative images of incomplete bony decompression. This means a reduction of additional costs which would arise with surgical revision. PMID- 18393164 TI - Off-midline Sinus Pericranii Associated with Ipsilateral Venous Anomaly: Case Report and Therapeutic Considerations. AB - A case of lateral sinus pericranii associated with ipsilateral venous anomaly is reported. No treatment was instituted in accordance with the parents' wishes. The role of MRI and cerebral angiography in the diagnostic workup is discussed. PMID- 18393165 TI - Progressive collapse of PMMA-augmented vertebra: a report of three cases. AB - Vertebroplasty using polymethylmethacrylate (PMMA) for augmentation is accepted as a safe and effective treatment for vertebral compression fracture. However, various complications related to PMMA vertebroplasty have recently been reported. We experienced three cases with progressive collapse of PMMA-augmented vertebra. Collapse progressed after augmentation in cases where PMMA conglomerated without contiguous bone interdigitation. A high viscosity of the PMMA preparation and vertebral body cavitory lesion may play a role in progressive vertebral collapse. To avoid this complication, bone cement should be injected sufficiently and permeate to contiguous bone to create strong support and anchorage. PMID- 18393166 TI - Gangliocytic paraganglioma of the cauda equina with significant calcification: first description in pediatric age. AB - A 17-year-old boy was referred with a 2-month history of low back pain and bilateral sciatica and difficulty in ambulation. Neurological examination found mild muscle weakness and diminished deep tendon reflexes in his right leg. Lumbar magnetic resonance imaging revealed an intradural-extramedullary tumor at the level of the L4, exiting through the right L4-L5 intervertebral foramen into the right psoas muscle. After total resection of the tumor, histopathological diagnosis revealed a gangliocytic paraganglioma. There are 184 paraganglioma cases reported at the lumbar region to date, and only 4 of them were in the pediatric age group. This report is the fifth paraganglioma case in the lumbar region and the first gangliocytic paraganglioma case in the pediatric age population. PMID- 18393167 TI - High cervical neurenteric cyst; acute post-traumatic rupture and respiratory failure: a case report. AB - The neuroenteric cyst is a rare developmental lesion originating from incomplete formation of the primitive neuroenteric canal. We report on the difficult management of an acute post-traumatic rupture of a cervical neuroenteric cyst. A 35-year-old male patient came to our observation after a spinal injury incurred during a football match. The patient immediately demonstrated tetraplegia and respiratory failure and was operated on. After two weeks the patient died of cardiopulmonary failure. High cervical neurenteric cysts, even those representing benign lesions, can produce dramatic results after a spinal injury. PMID- 18393168 TI - How Ramadan fasting affects caloric consumption, body weight, and circadian evolution of cortisol serum levels in young, healthy male volunteers. PMID- 18393169 TI - Lipocalin-2: development, analytical characterization, and clinical testing of a new ELISA. AB - Lipocalin-2 (also known as neutrophil gelatinase-associated lipocalin [NGAL]) has been described as a promising marker of metabolic syndrome associated with inflammation. The aim of our work was to develop an assay for the determination of lipocalin-2 in human serum and to investigate its levels in healthy volunteers and donors suffering from metabolic syndrome. We also conducted a pilot study on individuals with metabolic syndrome and on healthy probands and measured lipocalin-2 in these individuals. We developed and evaluated the sandwich ELISA method for the quantitative determination of human lipocalin-2 in serum samples. We measured blood pressure, waist circumference, serum cholesterol, triglycerides, HDL cholesterol, LDL cholesterol, insulin, glucose, creatinine, hs CRP, and adiponectin and calculated the BMI and Quicki insulin sensitivity index. In the study on 153 healthy volunteers, we showed that sex and age are not determinative for lipocalin-2 serum values. Furthermore, we tested 45 individuals with metabolic syndrome; values of lipocalin-2 did not differ (78.8 vs. 80.0 microg/l, p =0.56) from the data of healthy individuals from the first study. Neither group differed with regard to sex or age. Lipocalin-2 correlated with alanine aminotransferase (ALT) (r=-0.3, p<0.01) aspartate aminotransferase (AST) (r=-0.3, p<0.01), cholesterol (r=-0.21, p=0.047), creatinine (r=0.19, p=0.05), and high-sensitivity C-reactive protein (hs-CRP) (r=0.22, p=0.036). No significant correlation was found between serum lipocalin-2 and BMI, waist circumference, blood pressure, triglycerides, HDL, Quicki, or the number of metabolic syndrome components. When study patients with metabolic syndrome were further stratified according to the number of components of metabolic syndrome, serum concentrations of lipocalin-2 did not differ. The results presented demonstrate the analytical competence of the lipocalin-2 assay. However, we assumed that lipocalin-2 is not a routinely usable marker of metabolic syndrome or obesity. The association between serum lipocalin-2 and obesity or metabolic syndrome was not validated in our study. PMID- 18393170 TI - Contribution of gluconeogenesis and glycogenolysis to hepatic glucose production in acromegaly before and after pituitary microsurgery. AB - The diabetogenic effect of excess growth hormone (GH) such as that in acromegaly is well known. However, the contribution of the various components to hepatic glucose production (HGP) is not completely understood. In this study we evaluated insulin resistance, HGP, gluconeogenesis (GNG), and glycogenolysis (GLY) in five patients with acromegaly before and after pituitary microsurgery. Insulin resistance was estimated by the HOMA index. HGP was measured using a primed continuous (6,6- 2H2) glucose infusion, and GNG was measured from 2 H enrichment at carbons 2 and 5 of blood glucose on ingestion of 2H2O. The ratio of these enrichments equals the fractional contribution of GNG to HGP, and GLY was calculated as the difference between HGP and GNG. All measurements were performed after 12 hours of fasting. Levels of GH and IGF-I decreased, as did the HOMA index (p<0.05). HGP was reduced from 11.4 micromol/kg/min to 9.7 micromol/kg/min (p=0.032). GNG contributed most to HGP. GNG was unchanged, whereas GLY's fraction decreased 29% (p=0.056) postoperatively. This pilot study indicates that GNG is the main contributor to HGP and that GLY is more sensitive than is GNG to the insulin resistance existing in acromegaly. PMID- 18393171 TI - Second-generation thyrotropin receptor antibodies assay and quantitative thyroid scintigraphy in autoimmune hyperthyroidism. AB - Graves' disease (GD) is characterized by circulating TSH receptor antibodies (TRAb), and so-called hyperthyroid autoimmune thyroiditis (H-AIT) generally shows negative TRAb results with first-generation assays. However, a positive titer was observed in up to 13% of patients with euthyroid or hypothyroid autoimmune thyroiditis (AIT) by second-generation TRAb assays, and a larger increase is expected in hyperfunctioning forms. A thyroid 99mTc-pertechnetate uptake (TcTUs) cutoff of 2% previously was shown to accurately discriminate between GD and H AIT. Here we evaluated the relationship between second-generation TRAb assays and TcTUs in 139 patients with untreated autoimmune hyperthyroidism. An increase in TRAb levels was found in 114 of 139 patients (82%). All patients with TcTUs >2% and 66% of those with lower values had positive TRAb measurements. When the cutoff was increased to 5.9 U/l, positive TRAb occurred in 92.6% of those having higher TcTUs and in 1.4% of those with lower TcTUs. TRAb levels significantly increased in patients with TcTUs higher than 2% as compared with those having lower values, while no differences occurred in patients with TcTUs ranging from 0 to 1%, from 1.1 to 1.5%, or from 1.6 to 2%. A significant relationship between TRAb and TcTUs (as well as fT3, fT4, and thyroid volume) was found. TcTUs, fT4, fT3, and thyroid volume significantly increased in patients with positive TRAb (cutoff 1.5 U/l) compared with those testing negative. Our data indicate that a large proportion of patients with hyperthyroidism and hypoechoic thyroid, including those previously diagnosed as having H-AIT, actually have circulating TRAb. TRAb levels predicate the degree of iodine uptake (as measured by TcTUs) and thyroid hyperfunction. The role of baseline TRAb measurement by second generation assays to predict patients' outcome needs to be further evaluated and compared with TcTUs in prospective studies. PMID- 18393172 TI - Mitogenic effect of the insulin analogue glargine in malignant cells in comparison with insulin and IGF-I. AB - The aim of the study was to investigate if the insulin analogue glargine, with an increased affinity for the IGF-I receptor (IGF-IR), affects the cell growth to a larger extent than human insulin in malignant cells expressing IGF-IRs. The breast cancer cell lines MCF-7 and SKBR-3, and the osteosarcoma cell line SaOS-2 were used. Gene expression was determined by real-time RT-PCR and receptor protein quantified by ELISAs. Receptor phosphorylation was assessed by immunoprecipitation and Western blot. Mitogenic effect was determined as (3)H thymidine incorporation into DNA. The gene expression of insulin receptor (IR) varied between 4.3-7.5 x 10(-3) and the expression of IGF-IR between 7.7-147.7 x 10(-3) in relation to GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Insulin receptor and IGF-IR protein varied between 2.0-4.1 ng/mg protein and 2.0-40.4 ng/mg protein, respectively. The IGF-IR was phosphorylated by IGF-I at a concentration of 10(-10)-10(-9) M. All three polypeptides stimulated DNA synthesis in MCF-7, SKBR-3, and SaOS-2 cells. SaOS-2 cells were more sensitive to IGF-I than to insulin and glargine. MCF-7 cells were more sensitive to des(1 3)IGF-I than to IGF-I. In SKBR-3 and SaOS-2 cells, glargine tended to be more potent than human insulin to stimulate DNA synthesis. Our results suggest that glargine, compared to human insulin, has little or no increased mitogenic effect in malignant cells expressing IGF-IRs. PMID- 18393173 TI - A short acute octreotide test for response prediction of long-term treatment with somatostatin analogues in acromegalic patients. AB - The usefulness of the acute octreotide test in the selection of acromegalic patients for chronic somatostatin depot analogues treatment is controversial. The aim of the present study was to determine its accuracy for chronic response prediction, and the reliability of a short version of the classic 6-hour test. The data from 26 acromegalics (19 women, 7 men, mean age 52.6+/-13.1 years) studied with an acute octreotide test (6 hours sampling for GH measurement after octreotide 100 microg s. c.) were retrospectively analyzed. Eighteen of them followed chronic somatostatin depot analogues treatment for 12 months. GH nadir was always detected at 2 hours (mean decrease 75.9+/-24%). GH levels at 2 hours positively correlated with the other time-points (r(s) 0.97, 0.98, 0.97, 0.96 at 3, 4, 5 and 6 h, respectively; p<0.0001). During chronic treatment with maximal effective dose for 12 months, 61% of the patients achieved IGF1 <3 SD and 22% reached IGF1 <2 SD. GH nadir correlated with IGF1 decrease at 12 months (r(s) 0.76, p<0001). GH nadir of 9.2 ng/ml predicts IGF1 <3 SD with 82% sensitivity and 58% specificity (75% PPV, 67% NPV); for IGF1<2 SD, 75% sensitivity and 58% specificity are obtained for GH nadir 3.6 ng/ml, with 33% PPV and 89% NPV. Acute octreotide test reliably predicts response to long-term treatment; the short, 2 hour version is fully informative for therapeutic decisions in acromegalic patients. PMID- 18393174 TI - Induction and reversibility of insulin resistance in rats exposed to exogenous D fructose. AB - Long-term exposure of normal rats to a fructose-enriched diet or drinking water is currently used as an animal model for experimental insulin resistance. The present study deals with a comparison between rats given access to either a fructose-enriched diet or fructose-enriched drinking water. In both situations, a decrease in food intake and body weight gain, and the induction of insulin resistance with intolerance to D-glucose despite increased secretory response to the aldohexose of insulin-producing cells were documented. Moreover, the rats exposed to exogenous D-fructose displayed a lesser sensitivity to overnight fasting than control animals, in terms of the alteration of glucose homeostasis and reduction of the ratio between plasma insulin and D-glucose concentration. It is also shown that the fructose-induced insulin resistance, as assessed in a hyperinsulinemic-euglycemic clamp, represents a phenomenon reversed within 15-30 days after removal of the keto-hexose from the drinking water. PMID- 18393175 TI - [Lung neoplasms--CT screening for better early identification]. PMID- 18393176 TI - [Announcement--Quality of Life Prize 2008]. PMID- 18393177 TI - [Sleep apnea--cardiovascular and metabolic effects of CPAP therapy in adiposity]. PMID- 18393178 TI - [Allergology--increased risks for asthma and allergies with colonization of helicobacter]. PMID- 18393179 TI - [Lung neoplasms--ultrasonic endoscopy as a first diagnostic tool]. PMID- 18393180 TI - [Benign and malignant solitary pulmonary nodules--PET/CT for differential diagnosis]. PMID- 18393181 TI - [Evidence and technique for exercise training in patients with COPD]. PMID- 18393183 TI - [The voice(s) of pneumology]. PMID- 18393203 TI - [Evidence-based management of perinatal depression]. AB - Perinatal depression, which may occur from pregnancy to one year after childbirth, is recognized by the World Health Organization as a significant health issue affecting women. Depression during the perinatal period can have enormous consequences, not only affecting the health of the woman herself but also influencing her interaction with her children and other family members. This article introduces several depression screening tools and evidence-based nonpharmacological managements of perinatal depression. There are some fairly valid and feasible screening methods, among which routinely screening perinatal women with EPDS (Edinburgh Perinatal Depression Scale) or BDI (Beck Depression Inventory) in the primary care setting is practicable. A survey of the limited literature available reveals that interpersonal psychotherapy, cognitive behavior therapy and listening to music provide quantifiable depression amelioration effects for perinatal women. More scientific research moderated by women's life experiences and preferences should be conducted, however, and applied to improve women's health. PMID- 18393204 TI - [Using evidence-based nursing to explore the management of child fever]. AB - Fever is the most common problem in medical situations, but controversy still surrounds its benefit/ risk ratio for human beings. Fever is therefore always over-managed, and there is no consistency in management approaches to it. This article discusses fever management through the concept of evidence- based nursing, and provides a reference for clinical nursing. From the starting point of understanding the definition of fever and its physiological reactions, the benefit/risk ratio and management suggestion are discussed. Medical staff are expected to be able to use evidence in deciding approaches to clinical fever management, and to provide correct information with an appropriate attitude to the patient's family members, so that both medical staff and patients can enjoy consistent recognition of and management approaches to fever. PMID- 18393205 TI - [Cancer pain]. AB - The incidence of pain in patients with cancer is high. Pain is also one of the most feared and burdensome symptoms in cancer patients. Unrelieved pain has the negative effects of reducing daily functionality and quality of life. The main indicators of degree of pain and evaluators of how to handle that pain are markers of cancer pain. Complete records are the starting point for pain management. Improvements in pain management require an understanding of the views of patients, staff, health care providers and health care systems about pain assessment and reassessment, and about pain management. As regards the problem of pain education to improve cancer pain assessment and management, opioids are the main drugs for cancer pain management. When nurses care for cancer patients they need to know how to handle their side effects in order to ensure that cancer patients experience the maximum possible pain relief and comfort. PMID- 18393206 TI - [Symptom clusters in cancer patients]. AB - Cancer patients often experience multiple symptoms simultaneously. These symptoms can, either individually or jointly, exert significant impacts on patients' lives. Recently, the focus of symptom research has shifted from individual symptoms to symptom clusters. The purposes of the current paper are to review the studies on symptom clusters and to provide suggestions for clinical practice and research. The symptom cluster has been defined as two or more symptoms that are correlated. Research on symptom clusters can be classified into three types. The first type usually identifies, on the basis of researchers' experience or previous studies, a single symptom cluster containing three to four correlated symptoms. The second type focuses on using statistical methods to identify multiple symptom clusters. The third type uses statistical methods to classify patients in accordance with their symptom profiles. The sickness behaviors induced by pro-inflammatory cytokines found in animal studies have been reported to be similar to symptoms experienced by cancer patients. This finding suggests a common etiology for cancer symptom clusters. Studies have identified some patient factors that correlate with the severity of symptom clusters, and have reported that some symptom clusters may exert a negative effect on the patient. The literature suggests two approaches to managing symptom clusters: utilizing the phenomenon of symptom overlap or utilizing potentially common physiological mechanisms. This paper also discusses unresolved issues in symptom cluster research. PMID- 18393207 TI - [Job satisfaction and quality of life among hospital nurses in the Yunlin-Chiayi area]. AB - The purpose of this study was to explore job satisfaction and quality of life and their inter-relationships among nurses. A cross-sectional research design was applied. Participants were 1,020 nurses who had worked for over six months at seven hospitals in Yunlin and Chiayi counties. Nine hundred and eleven questionnaires were returned, a response rate of 89.3%. The questionnaire comprised three parts: demographic characteristics and work environment, quality of life, and job satisfaction. The Cronbach's alphas were .87-.94. Data were analyzed by SPSS/PC 13.0. The results showed: (1) Factors affecting job satisfaction were support managers, number of patients cared for during day time, health status, stress from changing units, religion, work stress, and working unit's suitability to one's interests (R(2) = 53.5%). (2) Factors affecting quality of life were job satisfaction, happiness of life, health status, work stress, and age (R(2) = 51.0%). (3) There was a positive correlation between job satisfaction and quality of life. Nursing managers should create better work environments to improve nurses' job satisfaction and facilitate their retention in the nursing profession. PMID- 18393208 TI - [Health related behaviors and associated factors among visitors to a health promotion center]. AB - The purpose of this study was to examine the health promoting behaviors and associated factors among patients who visited a health promotion center to receive nursing counseling. A cross-sectional survey and convenience sampling were applied in this study. Four hundred visitors to a medical center in northern Taiwan participated in the investigation. The results showed that: (1) The majority of participants were female, working, married, had a mean age of 41.5 years old, and were educated to college level. More than half of them had a religious belief, and most of them did not smoke, chew betel nut or drink alcohol; nearly one-third were overweight or obese. (2) Among the 40 items, there were nine behaviors categorized as high frequency, e.g. eating breakfast, maintaining good relationships etc. Four behaviors were categorized as low frequency, e.g. discuss with health personnel, check pulse rate when I do exercise, etc. (3) The demographic characteristics showed that those who worked, were married, more than 40 years old, and did not smoke had significantly higher scores on health promoting behaviors than their counterpart group. (4) The regression analysis showed that the higher the frequency of eating breakfast, the higher the age group, and the higher the educational level, and that unemployment explained the total variance of 14.6% in health promoting behavior. These findings will be an important reference for the implementation of health promoting programs for hospital visitors. PMID- 18393209 TI - [Preference for the attributes of home care service among primary family caregivers]. AB - This study uses direct scaled measurement to investigate the preference for the attributes of home care service among primary family caregivers. One hundred and fourteen effective questionnaires were collected from caregivers. The response rate was 100%. Data were analyzed with the SPSS software. Main findings include: (1) On average, the most valued product benefits were 'convenience', 'time savings', and 'financial savings'. (2)As regards the Carrier Product, the most needed items were 'assessment of the needs of physiological care', 'Instruction in tube care technique', 'oral instruction' about method of care, 'emergency handling' for health education, 'nursing personnel', 'psychological support' for spiritual care, and 'listening'. (3)With the Augmented Product: the most important items were 'quality of service' and 'image of organization'. This article analyzes the relationship between the personal attributes of principal caregivers and preferred residential service products. PMID- 18393210 TI - [Enhancing the capability of medical team to manage aggressive events in acute psychiatric wards]. AB - Incidences of violence in acute psychiatric ward can lead to not only facility destructions, but also mental, physical injuries and even medical disputes. As part of efforts to enhance medical team abilities to manage aggressive events, this study aimed to provide references for reducing both aggressive events and resultant damage. Over two-thirds (69%) of all unanticipated occurrences registered by our unit in 2003-2004 were classed as "aggressive events", i.e. there were 27 occurrences (0.09%) in which 0.04% resulted in staff injury. Events were mainly attributable to psychiatric symptoms, poor impulse control and interpersonal conflicts. For this study, we used several intervention methods, including categorizing patients by "risk of violence" rank, revising the hospital's standard operation processes for handling violence and revising the nursing rules to enhance nurse skills at managing violent events, countering patient violence, helping patients safely vent their anger and physical force, listening to relax music and conducting behavior modification. As a result, aggressive event prediction sensitivity increased from 56% to 100%, with successful prevention rates reaching 80%. The rate of aggressive event occurrence reduced from 0.09% to 0.06% and staff injuries decreased from 0.04% to 0.02%. Intervention methods employed were shown to be quite effective. If medical teams elsewhere enhanced their sensitivity and abilities to avoid aggressive events, injury and damages could be prevented and medical care quality enhanced. PMID- 18393211 TI - [Pain in elderly people with dementia]. AB - Aging is associated with an increased risk of cognitive impairment, such as Alzheimer's disease, vascular dementia and frontotemporal dementia. These diseases not only impair brain tissues and the nervous system, but also affect patients' verbal and non-verbal communicative ability. It is difficult, for instance, to assess pain in the cognitively impaired elderly because pain perception draws heavily on a patient's subjective interpretation and tolerance level. To seek an effective solution for pain detection in patients with dementia is therefore essential. From a systematic literature review covering the past decade, this paper presents a synopsis of the difficulty of pain assessment in the elderly with cognitive impairment, changes in the academic concept of pain, and explores factors resulting in obstacles to pain recognition. The obstacles to pain assessment derive mainly from three general factors. Firstly, classic definitions of early stage pain are inappropriate when applied to the elderly with cognitive impairment. Secondly, pain indicators are also unclear for this population. Thirdly, pain instruments and informants are ineffective in evaluating elderly patients with varying cognitive levels. To address these obstacles, a sound, multifaceted model of pain assessment for elderly patients with different severities of cognitive impairment is presented. On the basis of the above literature review, multiple methods for detecting aspects of pain in elderly people with cognitive impairment is recommended. To test the feasibility of the multidimensional model of pain assessment, further study is needed. PMID- 18393212 TI - [Mind mapping: a new tool for enhancing student learning strategy]. AB - With the rapid pace of development and reform in education learners face many challenges. Learning how to acquire skills and how to think are very important issues. The application of mind mapping can help students to enhance the creative thinking and problem-solving abilities of the whole brain. In other words, mind mapping is a visual or pictorial thinking method. This paper introduces the basic concept of mind-mapping, radiant thinking, the methods of mind-mapping, its rules of application, and examples of such application, to improve understanding and knowledge about mind-mapping. PMID- 18393213 TI - [Developments and trends in traditional medicine and complementary and alternative medicine]. AB - With the rapid development of society, medical science and technology, although quality of life is enhanced and life expectancy is prolonged, aging, environmental changes and health problems are unavoidable. More and more people, therefore, are concerned about their health and place high demands on medical care. As modern medicine cannot meet all such demands, other medical care systems emerge. Trends in the seeking of medical care show that people are inclined towards natural approaches, so attention is being paid once again to traditional medicine, as well as complementary and alternative medicine. Under the patient oriented concept, medical personnel have to recognize means of health care while thinking of the individualized and socioeconomic impacts. The purpose of this paper therefore is to provide medical personnel with information on the developments and trends in, knowledge and research with regard to traditional medicine as well as complementary and alternative medicine. PMID- 18393214 TI - [Clinical application of super-link system theory in spinal cord injury patients during rehabilitation stage]. AB - The purpose of this paper is to introduce Super-Link System Theory, which is a theory of spinal cord injury rehabilitation. This theory has been developed using the grounded theory research method. By explaining the procedure for establishing a super-link system the paper explains the complex structure of this theory. Super-Link System Theory emphasizes that rehabilitation nurses 'build up their interpersonal relationships' with clients, family caregivers, the interdisciplinary team, and the community, and attempt to 'establish links' among them. They know these links have to be made with appropriate 'timing', and must be able to access the appropriate people when necessary. Super-link systems include the following four links: link to client with spinal cord injury, link to family caregiver, link to interdisciplinary rehabilitation team, and link to community. It can enable rehabilitation nurses to provide a better quality of nursing care to clients and their family caregivers, as well as promote their professional position in the interdisciplinary rehabilitation team. PMID- 18393215 TI - [The implications and benefits of overseas nursing programs]. AB - In the era of globalization, universities in Taiwan regard internationalization of education as an important item on their educational agenda, and expansion of students' overseas educational opportunities as a strategy by which to globalize education. Overseas nursing study programs can enhance nursing students' cultural competence, help them understand different healthcare systems within different cultures and countries, and assist their personal development (enhancing, for example, self-understanding, independence, and leadership skills). Appropriate preparation for students who are willing to participate in overseas programs includes language preparation and cultural preparation. The design and planning of "overseas nursing programs", and related discussion and research remain in a developmental stage, and might therefore serve as a focus for industrious future teaching and investigation. PMID- 18393216 TI - [The nursing of a lower rectal cancer patient after sphincter-preserving surgery]. AB - The anal sphincter-preserving operation (ASP) has represented a significant improvement in surgical procedure for lower rectal cancer patients in recent years. Discussion has continued, however, on how to manage the reduced fecal function after such surgery. This case report describes how to help a patient with lower rectal cancer after ASP to overcome the problems due to changes in fecal function. Process recording was used to document the nursing process from Jan. 18 to Feb. 1, 2007. Data collected from interview, observation, and medical records, show that the patient revealed four vital nursing problems: fecal function disorder, impaired skin integrity, inappropriate regulation of diet, and powerlessness. After practicing pelvic floor exercises, adjusting her fecal behavior and modifying her food intake, the patient regained control of her fecal function and improved her quality of life. PMID- 18393217 TI - [Applying the family adjustment and adaptation response model to care of an AIDS patient's family]. AB - Discrimination against AIDS patients occurs in our society, not only influencing the patients but also restricting their usage of social resources. We report on a 31-year-old AIDS patient facing a family crisis because of an imbalance between the meanings, demands, and capabilities of his family. In this paper, we have applied the family adjustment and adaptation response (FAAR) model to assess this family, and identified three health problems, including (1) poor communication among family members, (2) deficiency in disease-related knowledge and skill, and (3) dysfunctional processing among the family. Throughout the care, we adopted a family-centered belief to communicate with the family to resolve their misunderstandings. We provided the family with information related to the disease to reduce their stress arising from caring for the patient. We also helped the family to utilize social resources by coordinating multi-disciplinary care. The family successfully restored a balance between meanings, demands, and capabilities. If we can apply this model clinically to understand the capabilities, demands and meanings of family existence as well as to help family members to develop their capabilities, reduce demands and recognize the positive meanings of family, we can help AIDS patients and their families to achieve positive adaptation. PMID- 18393219 TI - On the absence of mutations in nucleotide excision repair genes in sporadic solid tumors. AB - In general, stochastic tumors show genomic instability associated with the proliferation of DNA point mutations, that is, a mutator phenotype. This feature cannot be explained by a dysfunctional mismatch repair alone, and indicates that nucleotide excision repair (NER) and/or base excision repair should be suppressed. However, mutations in NER genes are not causally implicated in the oncogenesis of sporadic solid tumors, according to the Cancer Gene Census at http://www.sanger.ac.uk/genetics/CGP/Census/. This brings up an apparent paradox: how to explain the recurrent non-existence in NER genes of somatic mutations causally related to cancer? In a recent study, we have shown that the origin of point mutations in cancer cell genomes can be explained by a structurally conserved NER with a functional disorder generated from its entanglement with a disabled apoptosis gene network. In the present study, we further characterize NER gene network properties and show that it has a highly connected architecture. This feature suggests that the absence of mutations in NER genes in sporadic solid tumors is a result of their participation in many essential cellular functions. PMID- 18393218 TI - Structural evaluation of type 3 dopaminergic receptor gene (DRD3) in chronic anovulatory women. AB - Dopamine receptor type 3 (DRD3) expressed in the limbic system sites involved in the regulation of GnRH seems to play a role in neuroendocrine control. We hypothesized that women with chronic anovulation should show exacerbated secretion of prolactin (PRL) after thyrotropin-releasing hormone (TRH) stimulation test, having more chances for dopamine inhibitory dysfunction due to alterations in the structure of DRD3. The DRD3-coding region was evaluated in 60 women with chronic anovulation (35 without and 25 with hyperresponse of PRL after TRH stimulation), and in 34 controls. Statistically similar frequencies of homozygous AGC polymorphism (43.4 and 33.4%) and heterozygous polymorphism (33.4 and 47.9%) at position 9 were found in controls and patients, respectively. Homozygous GCG polymorphism at position 17 was identified in 3.4% of the patients, while heterozygosis occurred in 20.8% of the patients and in 6.6% of the controls. The novel 41563_41567delTAAGT polymorphism of DRD3 was identified in 14.7% of the controls and 8.6% of the women with chronic anovulation displaying hyperresponse of PRL after TRH stimulation. Alteration 41563_41567delTAAGT of DRD3 was not found in patients who did not show hyperresponse of PRL after TRH stimulation. Normal baseline and peak levels of PRL and thyroid-stimulating hormone were similar for women with and without 41563_41567delTAAGT in the DRD3 gene. It is concluded that the novel polymorphism in DRD3 identified in this study is not associated with the response of PRL to TRH stimulation in women with chronic anovulation. PMID- 18393220 TI - Cytogenetic and random ampliied polymorphic DNA analysis of Leptodactylus species from rural and urban environments (Anura, Amphibia). AB - Cytogenetic and random amplified polymorphic DNA analyses carried out in the species Leptodactylus podicipinus, L. ocellatus, L. labyrinthicus, and L. fuscus from rural and urban habitats of the northwest region of Sao Paulo State, Brazil, showed that the karyotypes (2n = 22), constitutive heterochromatin distribution and nucleolus organizer region (NOR) location did not differ between the populations from the two environments. The in situ hybridization with an rDNA probe confirmed the location of the NORs on chromosome 8 revealing an in tandem duplication of that region in one of the chromosomes of L. fuscus. DAPI showed that part of the C-band-positive heterochromatin is rich in AT, including that in the proximity the NORs in L. podicipinus and L. ocellatus. The molecular analyses showed that the two populations (urban and rural) of L. podicipinus and L. fuscus are similar from a genetic point of view. The urban and rural populations of species L. ocellatus and L. labyrinthicus showed differences in genetic structures, probably due to urbanization which interferes with the dispersion of those frogs. The marked differences observed between the two populations of L. ocellatus can be representing the cryptic condition of the species. Unweighted pair-group method of analysis and genetic distance analysis detected the genetic proximity between L. ocellatus and L. fuscus. The results indicate that there was no reduction in the genetic diversity in the populations from the urban environment; however, the survival of these frogs would not be guaranteed in the case of an increase in human impact especially for populations of L. labyrinthicus and L. ocellatus. PMID- 18393221 TI - Correlation of polymorphism C3435T of the MDR-1 gene and the response of primary chemotherapy in women with locally advanced breast cancer. AB - Primary chemotherapy is a useful strategy for the treatment of locally advanced breast cancer and therefore allows in vivo evaluation of the action of cytotoxic drugs and the possibility of accomplishing conservative breast surgeries, as well as the early treatment of metastasis. Mechanisms of resistance to the drugs include the action of protein associated with the efflux of drugs from the intracellular environment hindering their activity; one of the most studied proteins is P-glycoprotein codified by the MDR-1 gene. The presence of polymorphisms can determine different physiological actions of these proteins, intervening with the response of the drug's action. We evaluated the presence of single nucleotide polymorphism (SNP) C3435T of the MDR-1 gene and its correlation with the response to primary chemotherapy using the RECIST criteria. Forty-one Brazilian women with stages II and III breast cancer using the PCR-RFLP analysis were evaluated. Thirty-three patients with the SNP genotype (TT and CT) and eight patients with the wild genotype (CC) were found; there was no statistically significant correlation between the diverse genotypes and the clinical and pathological responses according to the Cramer correlation coefficient (V = 0.14). The parameters: nuclear and histological degree, and estrogens, progesterone and c-erb B2 receptors did not demonstrate a statistical correlation with the SNP C3435T. Patients with complete pathological response (12.5%) showed only the polymorphic genotype and not the wild genotype. The characteristics of miscegenation in our population could explain the absence of the characterization of a sub-group of individuals where the presence of the polymorphic genotype influenced the response to the primary chemotherapy. PMID- 18393222 TI - Agronomic and molecular characterization of introgression lines from the interspecific cross Oryza sativa (BG90-2) x Oryza glumaepatula (RS-16). AB - The reduced genetic variability of modern rice varieties (Oryza sativa) is of concern because it reduces the possibilities of genetic gain in breeding programs. Introgression lines (ILs) containing genomic fragments from wild rice can be used to obtain new improved cultivars. The objective of the present study was to perform the agronomic and molecular characterizations of 35 BC2F8 ILs from the cross O. glumaepatula x O. sativa, aiming to select high-yielding ILs to be used in rice-breeding programs. All 35 ILs were field evaluated in the season 2002/2003 in three locations and the 15 best performing ones were evaluated in the season 2003/2004 in five locations. In 2003/2004, six ILs (CNAi 9934, CNAi 9931, CNAi 9930, CNAi 9935, CNAi 9936, and CNAi 9937) showed the highest yield means and were statistically superior to the controls Metica 1 and IRGA 417. Molecular characterization of the 35 ILs was performed with 92 microsatellite markers distributed on the 12 rice chromosomes and a simple regression quantitative trait locus analysis was performed using the phenotypic data from 2002/2003. The six high-yielding ILs showed a low proportion of wild fragment introgressions. A total of 14 molecular markers were associated with quantitative trait loci in the three locations. The six high-yielding ILs were incorporated in the Embrapa breeding program, and the line CNAi 9930 is recommended for cultivation due to additional advantages of good grain cooking and milling qualities and high yield stability. The O. glumaepatula-derived ILs proved to be a source of new alleles for the development of high-yielding rice cultivars. PMID- 18393223 TI - Pattern of silver nitrate-staining during meiosis and spermiogenesis in testicular lobes of Antiteuchus tripterus (Heteroptera: Pentatomidae). AB - The pattern of silver nitrate (Ag)-staining differed among testicular lobes of Antiteuchus tripterus. In general, these differences are in regard to the number, size, shape, coloring intensity, and location of the stained bodies or masses, observed during meiosis and spermiogenesis. These characteristics were similar in lobes 1-3. Lobes 4-6, however, differed from each other and from lobes 1-3 as well. Because the Ag-staining method is specific for nucleolar organizing regions and nucleolar material, the observations in meiosis of lobes 1-3 suggested the presence of a single pair of nucleolar organizing region-bearing chromosomes in A. tripterus, as previously found in other Pentatomidae species. In general, the amount of Ag-stained material seen in meiosis of the testicular lobes 1-3 of A. tripterus is smaller than in the other lobes. The differences among lobes observed during spermiogenesis included a striking variation in morphology of the Ag-stained material found in the head and tail of the spermatids. Given that the key role of the nucleolar material is to participate in protein synthesis, interlobular variations seem to be related to the different functions attributed to each lobe (reproduction to lobes 1-3 and basically nutrition to lobes 4-6). To our knowledge, this is the first time that the nucleolar material was studied in each testicular lobe during spermatogenesis. The present observations encourage further studies since, in addition to being of basic biological interest, several Pentatomidae species are agricultural pests and added knowledge of their biology, mainly in reproduction, may be important for the development of control strategies. PMID- 18393224 TI - Prognostic value of TP53 Pro47Ser and Arg72Pro single nucleotide polymorphisms and the susceptibility to gliomas in individuals from Southeast Brazil. AB - The TP53 tumor suppressor gene codifies a protein responsible for preventing cells with genetic damage from growing and dividing by blocking cell growth or apoptosis pathways. A common single nucleotide polymorphism (SNP) in TP53 codon 72 (Arg72Pro) induces a 15-fold decrease of apoptosis-inducing ability and has been associated with susceptibility to human cancers. Recently, another TP53 SNP at codon 47 (Pro47Ser) was reported to have a low apoptosis-inducing ability; however, there are no association studies between this SNP and cancer. Aiming to study the role of TP53 Pro47Ser and Arg72Pro on glioma susceptibility and oncologic prognosis of patients, we investigated the genotype distribution of these SNPs in 94 gliomas (81 astrocytomas, 8 ependymomas and 5 oligodendrogliomas) and in 100 healthy subjects by the polymerase chain reaction restriction fragment length polymorphism approach. Chi-square and Fisher exact test comparisons for genotype distributions and allele frequencies did not reveal any significant difference between patients and control groups. Overall and disease-free survivals were calculated by the Kaplan-Meier method, and the log rank test was used for comparisons, but no significant statistical difference was observed between the two groups. Our data suggest that TP53 Pro47Ser and Arg72Pro SNPs are not involved either in susceptibility to developing gliomas or in patient survival, at least in the Brazilian population. PMID- 18393225 TI - Chromosome number and cytogenetics of Euphorbia heterophylla L. AB - Euphorbia heterophylla L. (Euphorbiaceae) is a herbaceous species of great economic importance due to its invasive potential and consequent damage to agriculture and pasture land. For the first time, we provide information on its chromosome number, morphology, and behavior of mitotic chromosomes. Seeds were germinated and submitted to four treatments to obtain metaphases: 0.5% colchicine for 2 to 5 h, at ambient temperature; 0.5% colchicine for 16 to 24 h; 0.0029 M 8 hydroxyquinoline (8-HQ) for 2 to 5 h at ambient temperature, and 0.0029 M 8-HQ for 16 to 24 h at 4 degrees C. The material was then fixed in methanol:acetic acid (3:1) and kept at -20 degrees C for 24 h. Roots were macerated in the enzyme solution of Flaxzyme (NOVO FERMENT)-distilled water (1:40) at 34 degrees C for 2 h and later fixed again. Chromosome preparations were obtained by the dissociation of the apical meristems. The best chromosome preparations were obtained with the use of 8-HQ for 21 h 30 min at 4 degrees C. E. heterophylla showed 2n = 28 chromosomes. The short arm of the largest pair of chromosomes of the complement (pair number 1) displayed a secondary constriction while the nucleolus was observed in the interphasic cell. Structural rearrangements were also observed in the E. heterophylla L. genome. The genomic instability associated with polyploidy may be the result of selection shaped by environmental adaptations and/or human-induced manipulation through agricultural practices. PMID- 18393226 TI - Detection of deletions and duplications in the Duchenne muscular dystrophy gene by the molecular method MLPA in the first Argentine affected families. AB - Deletions/duplications in the Duchenne muscular dystrophy (DMD) gene account for 60 to 70% of all alterations. A new technique, multiplex ligation-dependent probe amplification (MLPA), has been described that allows the detection of large genetic rearrangements by simultaneous amplification of up to 45 target sequences. The present article is based on the diagnosis of the first Argentine affected families by the application of MLPA. DNA samples from patients with and without a previous diagnosis were included. MLPA assays were performed according to manufacturer recommendations. Polymerase chain reaction and direct sequencing were performed when a single-exon deletion was detected. Results were analyzed using the Gene Marker v1.6 and Sequencing Analysis v5.2 software. In the samples with a previous diagnosis (as identified by short tandem repeat-polymerase chain reaction analysis), MLPA confirmed in some samples the same deletion and detected in others a larger deleted fragment. This enabled the prediction of the expected male phenotype. One deletion and one duplication were detected in patients without previous diagnosis. In this study, we investigated the applicability of MLPA in our country. Our results showed a 100% confirmation of the deleted fragments detected by short tandem repeat segregation analysis. Moreover, in some cases, the MLPA assay was able to refine the breakpoints involved. In addition, MLPA identified deletions/duplications in samples without previous diagnosis. In comparison to the available diagnosis strategies in Argentina, MLPA is less time consuming, and spans the complete coding region of DMD. The application of MLPA will improve the genetic diagnosis of DMD/Becker muscular dystrophy in our country. PMID- 18393227 TI - Genetic analysis of average annual productivity of Nellore breeding cows (COWPROD). AB - With the aim of estimating the coefficient of heritability of average annual productivity of Nellore cows (COWPROD), a data set from 24,855 animals with known pedigree was analyzed. COWPROD is defined as the amount (in kilograms) of weaned calves produced yearly by one cow during her remaining time in herd ignoring a fixed period of 365 days. COWPROD was calculated regarding three standards: a) based on the post-weaning weight from the calves ignoring any kind of adjustment (COWPROD_NAJ), b) adjusted weight for the fixed effects (COWPROD_AJFIX) and c) adjusted weight for the fixed effects and for the genetic merit of the sire (COWPROD_AJFIN). The obtained heritabilities were 0.15, 0.15 and 0.16 for COWPROD_NAJ, COWPROD_AJFIX and COWPROD_AJFIN, respectively. A complete set composed of 105,158 COWPROD records on 130,740 animals in pedigree was also analyzed for predicting the genetic merit of all animals in the data set and for the calculation of the genetic, phenotypic and residual trends. Ranking correlation was high for the adjusted and non-adjusted data, yet, for some of the animals, the difference among the genetic values was large. This would be an indication that it would be better to work always with the adjusted weaning weights. The genetic trend was positive, but was of small magnitude (0.26% of the trait average) and the residual trend was negative as a consequence of the large intensification of the production system, which has been occurring in the last years in the farms studied. The phenotypic trend was also negative and intermediate between the genetic and the residual ones. PMID- 18393228 TI - Effects of polymorphisms of LHR and FSHR genes on sexual precocity in a Bos taurus x Bos indicus beef composite population. AB - The purpose of the present research was to investigate the effects of polymorphisms of luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) genes, evaluated by polymerase chain reaction-restriction fragment length polymorphism in European-Zebu composite beef heifers from six different breed compositions. The polymorphism site analysis from digestion with HhaI and AluI restriction endonucleases allowed the genotype identification for LHR (TT, CT and CC) and FSHR (GG, CG and CC) genes. A high frequency of heterozygous animals was recorded in all breed compositions for both genes, except in two compositions for LHR. The probability of pregnancy (PP) at first breeding was used to evaluate the polymorphism effect on sexual precocity. The PP was analyzed as a binary trait, with a value of 1 (success) assigned to heifers that were diagnosed pregnant by rectal palpation and a value of 0 (failure) assigned to those that were not pregnant at that time. Heterozygous heifers showed a higher pregnancy rate (67 and 66% for LHR and FSHR genes, respectively), but no significant effects were observed for the genes studied (P = 0.9188 and 0.8831 for LHR and FSHR, respectively) on the PP. These results do not justify the inclusion of LHR and FSHR restriction fragment length polymorphism markers in selection programs for sexual precocity in beef heifers. Nevertheless, these markers make possible the genotype characterization and may be used in additional studies to evaluate the genetic structure in other bovine populations. PMID- 18393229 TI - [Blockade of the aberrant aggregation of alpha-synuclein in HEK293 cells induced by overexpression of wild-type alpha-synuclein by RNA interference]. AB - OBJECTIVE: To construct specific and effective RNA interference(RNAi) plasmid for alpha-synuclein gene and investigate RNAi blockade of the aberrant aggregation of alpha-synuclein in HEK293 cells induced by overexpression of wild-type alpha synuclein. METHODS: Hairpin RNAs for four target sites were designed to construct four RNAi plasmids pSYNi-1, pSYNi-2, pSYNi-3 and pSYNi-4, using plasmid pBSHH1 vector under the control of the H1 promoter. Western blot and reverse transcription-PCR(RT-PCR) were performed to screen the most specific and effective RNAi plasmid. After confirming the sequences of the plasmids, they were co-transfected into HEK293 cells with the recombinant plasmids alpha-synuclein pEGFP by using lipofectamin 2000. The aberrant aggregation of alpha-synuclein was measured by EGFP fluorescence and immunocytochemistry for alpha-synuclein. The inclusions in the cultured cells were identified with HE staining. RESULTS: By Western blot and RT-PCR, pSYNi-1 showed the most effective RNAi gene silencing effect (69.6%). After transfecting the RNAi plasmid, the aberrant aggregation of alpha-synuclein in HEK293 cells induced by overexpression of wild-type alpha synuclein was inhibited. The Lewy body-like inclusions were found in cytoplasm of cultured cells in control group, but disappeared in HEK293 cells cotransfected by pSYNi-1 and alpha-synuclein-pEGFP plasmid. CONCLUSION: RNAi can block the aberrant aggregation and Lewy body-like inclusion formation in cytoplasm of HEK293 cell induced by overexpression of wild-type alpha-synuclein. PMID- 18393231 TI - [Effect of interleukin-6 and calcium sensing receptor gene polymorphisms on bone mass accrual in Chinese adolescent girls]. AB - OBJECTIVE: To investigate the effect of interleukin-6 (IL-6) and calcium sensing receptor (CASR) gene polymorphisms on bone mass accrual in Chinese adolescent girls. METHODS: A total of 228 premenarche Chinese girls (9-11.5 years) were recruited for a 2-year study. Bone mineral densities (BMD) and Bone mineral contents (BMC) in the total body, total left hip including femoral neck, trochanter, intertrochanter and Ward's triangle area, and lumbar spine (L1-L4) were measured by DEXA. The -634C/G polymorphism of IL-6 gene was detected by PCR restriction fragment length polymorphism and A986S polymorphism of CASR gene was detected by allele-special mutagenically separated amplication-PCR. RESULTS: There were 176 available subjects when the 2-year study was completed. The 634C/G polymorphism of IL-6 gene and A986S polymorphism of CASR gene were significantly associated with bone mass accrual after adjusting the potential confounding factors. Girls with CC genotype of IL-6 -634G/C gene had higher percentage of increase in BMD of total body (P= 0.027) and femoral trochanter (P= 0.028) than those with CG+ GG genotypes. Girls with AA genotype of CASR gene had greater percentage of increase in L1-L4 lumbar spine BMC (P= 0.022) and Ward's triangle area BMD (P= 0.049) than their AS+ SS counterparts. Subjects with G allele of IL-6 gene in combination with S allele of CASR gene had less increase in BMC of femoral neck and L1-L4 lumbar spine. CONCLUSION: Adolescent girls carrying the G allele of IL-6 gene at -634C/G and S allele of CASR gene at A986S could potentially be the risk population of lower bone mass accrual. PMID- 18393230 TI - [Association of alpha-adducin and angiotensin converting enzyme gene polymorphisms with salt-sensitive hypertension and early renal injury]. AB - OBJECTIVE: To investigate the association between the alpha-adducin gene G460T, angiotensin converting enzyme (ACE) insertion/deletion (I/D) polymorphisms and salt-sensitive hypertension and early renal injury in Chinese people. METHODS: The case-control study was performed in 200 essential hypertension (EH) and 200 normal control subjects in China. The 200 EH patients were divided into salt sensitive(SS= 109) and non-salt-sensitive(NSS= 91) groups according to modified Sullivan's method. The genotypes of alpha-adducin gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The ACE genotypes were determined by PCR. The urine microalbum (Alb) in 200 EH subjects was measured by radioactive immunoassay. RESULTS: (1) A higher frequency of alpha-adducin gene G460T TT in EH patients was observed (P< 0.05). No significant difference of the ACE gene I/D polymorphism was found between the EH patients and normal control (P> 0.05). There were significant differences in the alpha-adducin gene TT genotype and combined genotype of TT+ II between SS and NSS subjects (P< 0.05). (2) The levels of urine Alb/Cr in SS patients were significantly higher than that in NSS patients (P< 0.05); in SS group, the levels of urine Alb/Cr in ACE II and alpha-adducin gene TT genotypes were higher than that in ACE ID, DD genotype and alpha-adducin gene GT and GG genotypes. The levels of urine Alb/Cr in the group of alpha-adducin gene TT+ ACE II combined genotype were higher than that in other combined genotypes (P< 0.05). CONCLUSION: The alpha-adducin gene TT genotype or combined with ACE II are significantly associated with SS hypertension. The alpha-adducin gene TT and ACE II genotypes might be genetic susceptibility factors to hypertension accompanying renal injury. PMID- 18393232 TI - [Analysis of human hair basic keratin 6 gene mutation in a Chinese Han family with monilethrix]. AB - OBJECTIVE: To identify the human hair basic kerat in 6 gene (hHB6) gene mutation in a mother and her daughter with monilethrix. METHODS: Clinical data were obtained by investigating the pedigree and examining the patients' hair and skin. Total genomic DNA of the family participants was isolated. All exons and exon intron boundaries of hHB6 were amplified by PCR. Mutation screening was carried out using direct DNA sequencing. Restriction fragment length polymorphism (RFLP) analysis was used to confirm the mutation, and to investigate if the mutation co segregated with the disease in the family and existed in normal controls. RESULTS: A heterozygous transition of c.1204G to A (p.E402K) of hHB6 was identified. RFLP analysis demonstrated that affected members carried the p.E402K mutation, but the unaffected members of the family and 150 unrelated normal Chinese Han nationality controls did not carry the mutation. CONCLUSION: This study identified the hHB6 gene mutation c.1204G to A (p.E402K) in a Chinese monilethrix family. The mutation was inherited from the mother to her daughter. The results emphasized the key role of hair keratin hHB6 in the pathogenesis of monilethrix and indicated that the common mutation of hHB6 was also a cause of monilethrix in Chinese. PMID- 18393233 TI - [Positive association of apolipoprotein B gene C7673T polymorphism with cerebral hemorrhage with family history]. AB - OBJECTIVE: To investigate the relationship between C7673T polymorphism of apolipoprotein B (apoB) and cerebral hemorrhage with family history (CHFH) in Chinese Han in Changsha, Hunan province. METHODS: Fifteen families of CHFH and 93 sporadic cerebral hemorrhage patients and 100 normal controls were collected. The C7673T polymorphism of apoB was analyzed by PCR-restriction fragment length polymorphism and direct DNA sequencing. The triglyceride(TG), total cholesterol(TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol(LDL-C) levels were examined by oxidase method. The serum level of lipoprotein (a) was determined by immune method. RESULTS: (1)The allele T frequencies of apoB C7673T polymorphism in cerebral hemorrhage patients with family history, first-degree relatives, second-degree relatives, third-degree relatives, the sporadic cerebral hemorrhage patients and the control group were 0.176, 0.136, 0.058, 0.048, 0.081 and 0.040, respectively. (2) The allele T frequencies of apoB C7673T polymorphism in CHFH patients and their first-degree relatives were significantly higher than that of the control group (P< 0.01, P< 0.01), while there was no significant difference among second-degree relatives, third-degree relatives and control group (P> 0.05). And the allele T frequency of apoB C7673T in CHFH patients was significantly higher than that of sporadic cerebral hemorrhage patients (P< 0.05). (3)In CHFH patients and sporadic cerebral hemorrhage group, the levels of TC and LDL-C of the TC genotype were significantly higher than those of the CC genotype, while the level of HDL-C in the TC genotype was significantly lower than that of the CC geneotype (P< 0.05). CONCLUSION: (1)The allele T of apoB C7673T polymorphism may be related to cerebral hemorrhage with family history. (2) The allele T of apoB C7673T polymorphism may increase the susceptibility of cerebral hemorrhage by changing blood lipid levels. PMID- 18393234 TI - [A novel mutation in the SEDL gene leading to X-linked spondyloepiphyseal dysplasia tarda in a large Chinese pedigree]. AB - OBJECTIVE: To identify the genetic defect in a four-generation pedigree with X linked recessive spondyloepiphyseal dysplasia tarda (SEDT) from Southwest China. METHODS: Linkage analysis with one panel of fluorescently labeled microsatellite markers on chromosome X and mutation screening of SEDL gene by direct sequencing were performed. RESULTS: Linkage between SEDT and Xp22.2-Xp23.1 was established with maximum LOD score of 3.82 (theta = 0) between DXS987 and DXS8051. Upon sequence analysis, a point mutation within exon 4 of the SEDL gene (c.239A to G) was found which resulted in substitution of histidine with arginine at codon 80 (His80Arg). CONCLUSION: A novel missense mutation (H80R) was identified for SEDL gene in the large Chinese SEDT pedigree. PMID- 18393235 TI - [The transmission disequilibrium analysis between neuronal nicotinic acetylcholine receptor alpha 7 subunit gene polymorphisms and schizophrenia]. AB - OBJECTIVE: To investigate the association between neuronal nicotinic acetylcholine receptor alpha 7 subunit (CHRNA7) gene and schizophrenia. METHODS: The three polymorphisms rs2337980, rs1909884, rs883473 in CHRNA7 gene were detected based on PCR and polyacrylamide gel microarray in 129 schizophrenic trios. The results of genotyping were analyzed by haplotype relative risk analysis based on haplotype(HHRR), transmission disequilibrium test(TDT) and hyplotype analysis. RESULTS: (1)The HHRR analysis suggested that there was significant differences in rs2337980 allele frequencies between schizophrenia group and dummy control group(P= 0.017); (2)In TDT test, there may be transmission disequilibrium between rs2337980 and schizophrenia, the heterozygous parents excessively transferred the C allele to patients (P= 0.021); (3)The haplotype between rs2337980 and rs1909884 as well as the hyplotype among rs2337980, rs1909884 and rs883473 may have significant association with schizophrenia (global P= 0.034; global P= 0.027), the T-C and T-C-T hyplotype may have transmission disequilibrium with schizophrenia. CONCLUSION: There may be association between CHRNA7 gene polymorphisms and schizophrenia, the variant allele T in rs2337980 may have a protective effect to schizophrenia. PMID- 18393236 TI - [Effects of histone acetylation on differentiation of mesenchymal stem cells]. AB - OBJECTIVE: To explore the effects of histone acetylation modification on the regulation of mesenchymal stem cells (MSCs) differentiation into cardiomyocytes in myocardium microenvironment, by detecting the acetylase activity, expression of histone acetylation gene Gcn5 and myocardial gene GATA4 in myocardium transplanted by MSCs transfected with plasmid ZJ3. METHODS: Extracted the plasmid containing short hairpin RNA (shRNA) against Gcn5 gene (ZJ3) and transfected it into MSCs. After 24 hours, the MSCs were transplanted into the rat myocardium tissues and the acetylase activity, the expression of acetylation gene and myocardium development gene were detected after two weeks. RESULTS: The acetylase activity in the experimental group was significantly lower than all control groups; Gcn5 and GATA4 expression in myocardium of experimental group had significant reduction compared with the control groups. CONCLUSION: Inhibition of histone acetylation in the MSC cells can inhibit the transcription process of specialized myocardium in the myocardium microenvironment. The result establishes foundation for researches of histone acetylation and mechanisms of MSCs differentiation. PMID- 18393237 TI - [Detection of adenomatous polyposis coli gene mutations in 31 familial adenomatous polyposis families by using denaturing high performance liquid chromatography]. AB - OBJECTIVE: To analyze the adenomatous polyposis coli (APC) gene mutations in familial adenomatous polyposis (FAP) in Chinese. METHODS: DNA was extracted from blood samples taken from 31 FAP families, and all exons of the APC gene were amplified with touch-down PCR. APC gene mutations were screened by denaturing high performance liquid chromatography followed by sequencing if abnormal profile was detected. RESULTS: Twelve categories of APC gene mutations were found in 15 FAP families (48.39%) including 4 novel mutations in coding region and 3 mutations in introns. The 4 novel mutations in coding region were frameshift mutations and located in codons 255, 677, 1192 and 1403 respectively. Most mutations were clustered in exon 15 of APC gene leading to frameshift and accounted for 86.67%. Others were nonsense mutations (13.33%). CONCLUSION: The mutation rate of the APC gene in this group of Chinese FAP families was about 48.39%, and 4 novel mutations were detected. Frameshift mutation was the major mutation type in Chinese FAP and mainly located in exon 15. PMID- 18393238 TI - [Association of TP53 gene polymorphisms with genetic susceptibility to liver metastases of colorectal cancer]. AB - OBJECTIVE: To investigate the possible association between the single nucleotide polymorphisms (SNPs) (C-8343G, C-1863T and R72P) in TP53 gene and susceptibility to liver metastases of colorectal cancer (CRC) in a Chinese population. METHODS: The genotypes of each SNP in TP53 gene were determined by either TaqMan assays or PCR-based restriction fragment length polymorphism (RFLP) method in 121 colorectal cancer patients with liver metastases and sex-, age-matched 280 cases with nonmetastatic CRC as a control. Immunohistochemical staining for P53 was performed on paraffin-embedded sections. Odds ratios (ORs) for colorectal liver metastases and 95% confidence intervals (CIs) from unconditional logistic regression models were used to evaluate relative risks. RESULTS: No significant association of C-8343G or C-1863T with colorectal liver metastases risk was observed. However, the R allele of the TP53 R72P polymorphism was more frequently found in metastatic cases than in nonmetastatic cases (P= 0.037). When compared with PP homozygotes, the ORs of metastases for RP heterozygotes was 2.21 (95% CI: 1.13-4.33), for RR homozygotes was 2.26 (95% CI: 1.03-4.94), and for carriers of the 72R allele (RP or RR genotype) was 2.22 (95% CI: 1.16-4.26). Stratified analysis indicated that carrying the 72R allele had a more pronounced increase in colorectal liver metastases risk among patients with positive P53 expression tumors (OR= 3.28, 95% CI: 1.21-8.88), whereas no significantly increased metastases risk was found in patients with negative P53 expression tumors (OR= 1.37, 95% CI: 0.52-3.62). CONCLUSION: The R allele of the TP53 R72P polymorphism may contribute to the etiology of liver metastases in CRC patients, particularly among those with positive P53 expression tumors. Both TP53 C-8343G and C-1863T may be not associated with colorectal liver metastases risk. PMID- 18393239 TI - [A novel mutation of the PAX6 gene identified in a northeastern Chinese family with congenital aniridia]. AB - OBJECTIVE: To identify the mutation of the PAX6 gene in a northeastern Chinese family with aniridia. METHODS: Three aniridia patients from the family were undergone full ophthalmologic examinations. Genomic DNA was prepared from venous leukocytes from these three patients, five non-carriers in the family as well as 100 healthy normal controls. The coding regions of PAX6 gene were analyzed by PCR amplification, single-strand conformation polymorphism and direct DNA sequencing. RESULTS: The sequencing result revealed one novel PAX6 mutation in the three patients with familial aniridia. The mutation is a 9 base pair(bp) deletion in exon 5 (c.483del9) that results in a putative PAX6 protein with in-frame deletions of aspartic acid, isoleucine and serine at the amino acids 41-43. CONCLUSION: A PAX6 gene mutation beyond the existing spectrum of mutations has been identified in a northeastern Chinese family with aniridia. The genetic analysis suggests that the novel mutation in the PAX6 gene may be the cause of the classical aniridia phenotype. PMID- 18393240 TI - [Calculation of paternity index for paternity testing with considering mutation]. AB - OBJECTIVE: To formulate recommendations in calculation of paternity index in paternity testing under considering mutations. METHODS: Different formulas under considering mutations were developed according to Brenner method. RESULTS: Different formulas under considering mutations were obtained. Both true exclusion and false exclusion of paternity were easily distinguished using these formulas when the genetic pattern was inconsistent with paternity. CONCLUSION: The scientific evidence for paternity testing can be obtained using these formulas under considering mutations when both the combined probability of exclusion and the paternity index meet the threshold values. However, when either the combined probability of exclusion or the paternity index can not meet the threshold values, more genetic markers should be added. PMID- 18393241 TI - [Study on the expression of survivin mRNA and protein in nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the relationship between survivin mRNA and protein expression and nasopharyngeal carcinoma (NPC). METHODS: Survivin mRNA and protein were detected by in situ hybridization and immunohistochemical S-P staining respectively. RESULTS: Among 64 cases of NPC, 42 cases (65.6%) were positive for survivin mRNA expression, 30 cases (46.9%) had high expression level. In 46 cases (71.9%) of NPC were positive for survivin protein expression, and 38 cases (59.4%) had high expression level. In 22 cases of NPC with detailed clinical information, the positive expression rates of survivin mRNA and protein in stages III+ IV of NPC were 66.7% and 61.1% respectively, which were higher than those in stages I+ II of NPC (50.0% and 50.0% respectively). There was no significant difference between survivin mRNA and protein expression regarding age or gender of NPC patients (all P> 0.05). The positive expression rates of survivin mRNA and protein in chronic nasopharyngitis group were 33.3% and 23.3% respectively, which were lower than those in NPC group (chi (2)= 12.04, P< 0.01 and chi (2)= 19.57, P< 0.01, respectively). In 64 cases of NPC, 36 cases were positive for both survivin mRNA and protein, and the expression of survivin mRNA and protein showed positive correlation (phi = 0.43). CONCLUSION: The expression of survivin gene may play some roles in the pathogenesis of NPC. Detection of survivin mRNA and protein will be helpful for diagnosis, clinical staging and prognosis of NPC. PMID- 18393242 TI - [Association of three single nucleotide polymorphisms of the E-cadherin gene with susceptibility to epithelial ovarian carcinoma]. AB - OBJECTIVE: To investigate the association of three single nucleotide polymorphisms (SNPs), i.e. -160C/A, -347G/GA and 3'UTR+ 54C/T, in the promoter region of E-cadherin gene (CDH1) with the risk of epithelial ovarian carcinoma. METHODS: The SNPs of CDH1 gene were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 207 epithelial ovarian carcinoma patients and 256 unrelated healthy women; immunohistochemistry was used to measure the level of CDH1 in different genotypes of 3'UTR+ 54C/T locus. RESULTS: There were no significant difference in the frequencies between patients and control women in the two loci (-160C/A, -347G/GA) of CDH1 gene (P> 0.05). However, there was significant difference in the frequency of CDH1 3'-UTR+ 54C/T genotypes (CC, CT and TT) between patients and controls (P< 0.05). The frequencies of the CC genotype and the C allele in patients (65.2%, 89.1%) were significantly higher than that in controls (52.7%i64.5%) (P< 0.01). The CC genotype significantly increased the risk to epithelial ovarian carcinoma, with adjusted odds ratio of 1.85 (95%CI= 1.27-2.69). In the ovarian tissues of patients, the expression of CDH1 from CC genotype was significantly lower than that with the other genotypes (CT+ TT) (P< 0.05). The -160C/A and -347G/GA polymorphisms were in linkage disequilibrium (D'= 0.999582) by analyzing with 2LD software. The -160A/-347GA haplotype was only observed in patients (5.1%), the 160C/-347GA haplotype decreased susceptibility to epithelial ovarian carcinoma, with adjusted odds ratio of 0.66 (95%CI= 0.45-0.96). CONCLUSION: CDH1 -160C/A and -347G/GA polymorphisms were not associated with the risk of epithelial ovarian carcinoma. However, the haplotype may have impact on the risk of epithelial ovarian carcinoma. The CC genotype of 3'-UTR + 54CT may be a potential risk factor for the epithelial ovarian carcinoma. PMID- 18393243 TI - [Deficient mRNA expression of specific protein 3 gene in peripheral blood mononuclear cells from patients with multiple sclerosis]. AB - OBJECTIVE: To characterize the deficiency of the mRNA expression of specific protein (SP3) gene in peripheral blood mononuclear cells (PBMCs) from Chinese patients with multiple sclerosis (MS) and study its correlation with the disease phenotypes. METHODS: Fifty-six patients with definite MS were collected and total RNA was extracted from their PBMCs. Specific primers corresponding to SP3 gene were designed and the mRNA expression of SP3 gene was detected by reverse transcriptase-PCR (RT-PCR) method. The deficiency of SP3 expression was compared among MS patients, irrelevant disease group and normal controls. RESULTS: Of the 56 MS cases, 23 (41.1%) were SP3-deficient. In contrast, the frequency of SP3 deficiency in normal subjects and irrelevant disease controls was 8.6% (5/35) and 14.3% (4/27), respectively. The frequency of the SP3-expression deficiency in MS patients was significantly higher than that in both control groups (P< 0.01). Within the MS cases, the scores of expanded disability status scale (EDSS) in the SP3-expressing subjects were significantly different from that in the SP3 deficient ones in the stable, but not in the active, phase of MS (P< 0.05). CONCLUSION: Author's observation suggested that deficient expression of SP3 gene occurs in Chinese MS patients, and that the SP3 expression may correlate with the clinical manifestations of MS and play roles in its immunological pathogenesis. PMID- 18393244 TI - [Molecular cytogenetic studies of 25 males with azoospermia]. AB - OBJECTIVE: To investigate the clinical correlation of chromosome abnormalities and microdeletion in azoospermic factor (AZF) region on Y chromosome in 25 patients with azoospermia. METHODS: Chromosome analyses were performed by using chromosome GTG-banding, Q-banding, fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) for AZF region on chromosome Yq. RESULTS: Seven cases showed abnormal chromosome karyotype (28%). In 8 azoospermic patients tested, 2 showed microdeletions of AZFb (SY127, SY134)+ AZFc (SY254, SY255) and AZFc(SY243, SY158) on chromosome Yq, respectively. CONCLUSION: Chromosome abnormalities and AZF microdeletion are major cause of azoospermia leading to male infertility; male with azoospermia and infertility should be referred to cytogenetic diagnosis by using chromosome GTG-banding, Q-banding after ruling out clinical factors including testopathy, obstructive azoospermia, and abnormalities in incretion and immune system. FISH or PCR analysis for AZF region on chromosome Yq should be done for the patient with azoospermia if Q-banding indicates the deletion above Yq12 region. It is of essential importance to provide precise diagnosis in genetic counseling for further clinical treatment. PMID- 18393245 TI - [Analysis of BRCA2 gene mutations among familial and/or early-onset breast cancer patients in eastern Shandong of China]. AB - OBJECTIVE: To investigate the prevalence of BRCA2 gene mutations among familial and/or early-onset breast cancer patients in eastern Shandong of China. METHODS: Fifty-two familial and/or early-onset breast cancer patients from unrelated family were analyzed. Genomic DNA was collected from the peripheral blood mononuclear cells, the coding sequences and exon-intron boundaries of BRCA2 gene were screened using denaturing high performance liquid chromatography (DHPLC), and the abnormal fragments were confirmed with direct DNA sequencing. RESULTS: Three mutations (5.8%) in BRCA2 gene were identified. They were 2001del TTAT, 4099C to T and 5873C to A. To our knowledge, all of them were firstly found in Chinese population. Furthermore, all the three mutations (12%) were identified in familial breast cancer patients, and none was in the early-onset patients. CONCLUSION: BRCA2 may play an important role in the familial breast cancer in eastern Shandong Chinese population, but not in the early-onset breast cancer. It is necessary to give genetic test to familial breast cancer patients in this population. PMID- 18393246 TI - [Germline mutation of adenomatous polyposis coli gene in Chinese patients with familial adenomatous polyposis]. AB - OBJECTIVE: To explore the characteristics of adenomatous polyposis coli (APC) gene germline mutations in Chinese patients with familial adenomatous polyposis (FAP). METHODS: Eighteen members from nine FAP pedigrees were studied by using multiplex ligation-dependent probe amplification(MLPA) to detect large fragment deletion of APC gene. Then, PCR were performed to amplify all exons of APC gene for mutation screening by denaturing high performance liquid chromatography (DHPLC). When abnormal elution profile of DHPLC was found, DNA sequencing was performed to determine the mutations. RESULTS: Mutations were identified in three pedigrees among the nine pedigrees. They were c.3184_3187 del CAAA in pedigree 2, c.5432C to T in pedigree 4 and c.3925_3929 del AAAAG in pedigree 9 respectively. Among them, c.5432C to T was novel. CONCLUSION: APC gene germline mutations can cause the development of FAP. The FAP patients without APC gene germline mutations could be caused by other mechanisms. PMID- 18393247 TI - [Positioning analysis of the Yq11 deletion in a patient with azoospermia]. AB - OBJECTIVE: To locate the deletion region of an azoospermic patient with a large deletion on his Y chromosome long arm. METHODS: Multiplex polymerase chain reaction was used to amplify fifteen sequence tagged sites (STS), namely sY84, sY86, sY87 in AZFa, sY102, sY117, sY118, sY119, sY115, DYS132, DYS383, sY1015, sY121, sY125, sY127, sY129 and sY134 in AZFb, sY152 in AZFd, sY1258, sY1291, sY254, sY255, sY158 and sY1201 in AZFc, and sY160 in Yq12. RESULTS: Only sY84, sY86, sY87, sY102, sY117, sY118, sY119, sY115 and DYS132 could be amplified while the others were negative. The breakpoints were found to locate in an area between AZFb sY115 and DYS383 spanning 8577. CONCLUSION: This study provided the exact breakpoints on Y chromosome AZF region in the patient. PMID- 18393248 TI - [Analysis of acyl-coenzyme A: cholesterol acyltransferase 1 polymorphism in patients with endogenous hypertriglyceridemia in Chinese population]. AB - OBJECTIVE: To investigate the polymorphism of acyl-coenzyme A: cholesterol acyltransferase 1(ACAT1) gene and its relationship with endogenous hypertriglyceridemia (HTG) in Chinese population. METHODS: A total of three hundred and seventy-two subjects (105 endogenous hypertriglyceridemics and 267 healthy controls) from a population of Chinese Han nationality in Chengdu area were studied using PCR-restriction fragment length polymorphism (RFLP). RESULTS: The frequency of C allele in normal Chinese at rs1044925 locus was 0.137, which was lower than that reported in the population of central and Southern Europe (0.354) (P< 0.05). The frequency C allele was 0.153 in HTG group. No significant difference between normal control and HTG group. In control group, subjects with genotype AA had a higher serum mean concentrations of low density lipoprotein cholesterol (LDL-C) and non-high density lipoprotein-cholesterol(nHDL-C) when compared with those of C allele carriers (AC and CC genotype carriers), respectively [(3.25+/- 0.68) mmol/L vs (3.03+/- 0.87) mmol/L, P< 0.05; (3.80+/- 0.71) mmol/L vs (3.23+/- 0.82) mmol/L, P< 0.05]. In HTG group, subjects with genotype AA had a higher high density lipoprotein-cholesterol (HDL-C) level compared with those of C allele carriers [(1.00+/- 0.28) mmol/L vs (0.87+/- 0.17) mmol/L, P< 0.05]. CONCLUSION: These results suggest that rs1044925 polymorphism in ACAT1 gene is not only associated with serum LDL-C and nHDLC levels in healthy Chinese subjects in Chengdu area, but also with HDL-C level in subjects with endogenous hypertriglyceridemia in this population. PMID- 18393249 TI - [Association of the XRCC1 and hOGG1 polymorphisms with the risk of laryngeal carcinoma]. AB - OBJECTIVE: To evaluate the association between the polymorphisms of X-ray repair cross complementing group 1 (XRCC1) and human 8-oxoguanine glycosylase I (hOGG1) gene and the risk for laryngeal carcinoma. METHODS: This is a case-control study comprised of two groups: 72 patients with laryngeal squamous carcinoma, and 72 controls without laryngeal carcinoma. The PCR-restriction fragment length polymorphism method was used to analyze the XRCC1-Arg399Gln, hOGG1-Ser326Cys polymorphisms. RESULTS: The frequencies of XRCC1-399Arg/Gln+ Gln/Gln and hOGG1 326Ser/Cys+ Cys/Cys genotypes in the case group were higher than that of the control group(P< 0.05). There was a 3.37-fold or 2.54-fold increased risk of laryngeal carcinoma for individuals carrying XRCC1-399Arg/Gln+ Gln/Gln or hOGG1 326Ser/Cys+ Cys/Cys genotypes, compared with subjects carrying XRCC1-Arg/Arg or hOGG1-Ser/Ser genotype, respectively. No statistically significant differences were found between the smoking group and non-smoking group for risk of laryngeal carcinoma. CONCLUSION: The amino acid replacement of XRCC1-399Arg to Gln and hOGG1-326Ser to Cys might lead to an increased risk of laryngeal carcinoma. The study demonstrated the positive association between the polymorphisms of XRCC1 and hOGG1 genes and laryngeal carcinoma. PMID- 18393250 TI - [The cytogenetic features of Burkitt leukemia]. AB - OBJECTIVE: To evaluate the incidence of chromosomal abnormalities in Burkitt leukemia (BL). METHODS: Conventional cytogenetics (CC) was carried out to detect the karyotypes. Meanwhile, three color interphase fluorescence in situ hybridization (FISH) was used to detect the t(8;14)(q24;q32) in 17 newly diagnosed BL. RESULTS: The results showed that the incidence of chromosomal abnormalities was 41.2% (7/17) by CC technique, including one of each for simple t(8;14)(q24;q32), complex chromosomal abnormality containing t(8;14)(q24;q32), t(8;22)(q24;q11), the complex chromosomal abnormality of t(12;22)(?;?), t(2;13)(?;?) with + 12, and one with two marker chromosomes. FISH method detected eight cases of t(8;14)(q24;q32), including the two detected by CC technique. Five samples (5/8) showed 2A1G1O2F (two blue, one green, one orange and two yellow signals in interphase nuclei), while three samples (3/8) showed 2A1G1O1F(two blue, one green, one orange and one yellow signals in interphase nuclei). CONCLUSION: Two different breakpoints have been identified on the c-Myc locus on 8q24. Interphase FISH was more sensitive in detecting t(8;14)(q24;q32), and it is an important complement to CC. It should be used as a routine method for diagnosis of BL. PMID- 18393251 TI - [FGFR2 gene mutation in a family with Crouzon syndrome and a sporadic Crouzon syndrome patient]. AB - OBJECTIVE: To detect the gene mutation of fibroblast growth factor receptor (FGFR2)in a Crouzon syndrome family and a sporadic patient. METHODS: The genomic DNA from 10 members in the Crouzon syndrome family, as well as a sporadic patient, was extracted. Then exons 8 and 10 of FGFR2 gene and their flanking sequences were amplified by polymerase chain reaction. Some of the family members were studied by only amplifying exon 8. Finally, the PCR products were purified and sequenced. RESULTS: The G to T transversion mutation (heterozygote) at nucleotide 833 in exon 8 of FGFR2 (C278F), was found both in the patients of the family and the sporadic patient. CONCLUSION: FGFR2 gene mutation is responsible for the pathogenesis of Crouzon syndrome in these patients. PMID- 18393252 TI - [Mutation screening of MLH1 and MSH2 genes in two Chinese families with hereditary nonpolyposis colorectal cancer]. AB - OBJECTIVE: To identify the MLH1 and MSH2 gene mutation in two hereditary nonpolyposis colorectal cancer (HNPCC) families. METHODS: Polymerase chain reaction and DNA sequencing were used to screen for MLH1 and MSH2 gene mutation, and PCR-restriction fragment length polymorphism and DNA sequencing were performed to confirm the mutation. RESULTS: By DNA sequencing, a novel mutation of c.243_244insA located at the exon 3 of MLH1 gene was detected in family A, while c.1215_1218dupCCGA mutation located at the exon 7 of MSH2 gene was detected in family B. These two mutations can cause the formation of premature proteins. CONCLUSION: The novel mutations c.243_244insA in MLH1 gene and c.1215_1218dupCCGA in MSH2 gene were the disease-causing mutations in the two HNPCC families. PMID- 18393253 TI - [Association of CETP and CRP gene polymorphisms with non-valvular atrial fibrillation in Chinese Han population]. AB - OBJECTIVE: To study whether the polymorphisms of TaqIB of cholesteryl transfer protein (CETP) gene and 1444C/T of C reactive protein (CRP) gene are associated with non-valvular atrial fibrillation in the Chinese Han population. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was used to detect the distribution of genotypes of CETP TaqIB and CRP 1444C/T in 147 patients with non-valvular atrial fibrillation and 147 control subjects in Chinese Han population. RESULTS: (1) The distribution of CETP TaqIB and CRP 1444C/T genotypes was in Hardy-Weinberg equilibrium. (2) A statistically significant difference between patients and controls for CETP TaqIB (P= 0.005, OR= 0.614, beta = -0.488) and CRP 1444C/T (P= 0.003, OR= 2.428, beta = 0.887) was observed. (3) In female group, significant difference was observed in smoking, CETP TaqIB and CRP 1444C/T polymorphisms. And in male group, significant difference was observed in body mass index and CETP TaqIB polymorphisms. CONCLUSION: (1) These results suggest that CETP TaqIB (B2 allele as protective factor) and CRP1444C/T (T allele as risk factor) genetic polymorphisms may be associated with the non-valvular atrial fibrillation in the Chinese Han population. (2) Smoking and CRP1444T single nucleotide polymorphism may induce hereditary susceptibility to non-valvular atrial fibrillation in female. Obesity may induce hereditary susceptibility to non-valvular atrial fibrillation in male. PMID- 18393254 TI - [Establishing a single-tube fluorescent bidirectional PCR method to detect the 609C/T polymorphism of NQO1 gene]. AB - OBJECTIVE: To develop a single-tube fluorescent bidirectional PCR method to detect the 609C/T polymorphism of NAD(P)H: quinone oxidoreductase 1 (NQO1) gene. METHODS: Two primers of NQO1 gene C609T locus were designed. Using these primers, a SYBR Green I fluorescent bidirectional PCR, combined with melting curve analysis of the PCR products, were optimized to differentiate the 609C/T polymorphisms in 191 samples of human genomic DNA. The accuracy of the fluorescent bidirectional PCR was validated by the classical method of PCR restriction fragment length polymorphism(RFLP) in 62 of these 191 samples. RESULTS: In the 62 samples, the genotypes determined by the fluorescent bidirectional PCR were 100% consistent with the ones by the PCR-RFLP. The frequencies of genotypes of homozygous wild-type (CC), heterozygous (CT), and homozygous mutant (TT) were 28%, 50%, and 22%, respectively, in the 191 samples. CONCLUSION: The single-tube fluorescent bidirectional PCR method established here provides a simple, rapid, accurate and inexpensive assay to determine the 609C/T polymorphism of NQO1 gene. The assay is suitable to detect the single nucleotide polymorphism in large-scale samples. PMID- 18393255 TI - [Association of PAI-1 gene polymorphism with prognosis of coronary artery disease]. AB - OBJECTIVE: To investigate the association of the 4G/5G polymorphism located in the promoter region of plasminogen activator inhibitor-1(PAI-1) gene with prognosis of coronary artery disease (CAD) in Chinese Hans. METHODS: One hundred and fifty five patients with CAD and 190 unrelated healthy control individuals were included in the study. The 4G/5G polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. A follow-up survey of major adverse cardiovascular event (MACE) and analysis of the relationship between the severity of coronary vessels and PAI-1 gene polymorphism were carried out. RESULTS: (1) The frequency of 4G/4G genotype of PAI-1 gene was higher in CAD patients than in controls (58/155, 37.42% vs 52/190, 27.37%, P< 0.01). (2) The frequency of 4G/4G genotype of PAI-1 in patients with MACE was higher than that in patients without MACE (40/81, 49.38% vs 18/74, 23.42%; P< 0.01). (3) The frequency of 4G/4G genotype in patients with multivessel disease was higher than that in patients with single-vessel disease (30/47, 44.77% vs 9/37, 24.32%; P< 0.05). CONCLUSION: The 4G/5G polymorphism located in the promoter region of PAI-1 gene was associated with prognosis of CAD patients, and may be regarded as a biomarker of the severity of the involved vessels. PMID- 18393256 TI - [Responsive changes to MPTP in rats with DAT inhibited by antisense oligonucleotides]. AB - OBJECTIVE: To investigate the inhibitory effect of antisense oligonucleotides (ODN) on dopamine transporter (DAT) in rats and observe the response of the rats to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). METHODS: The cannula was implanted in the substantia nigra compacta under a rat stereotaxic device, through which drugs were used. The rats with successful operation were divided randomly into four groups, and received injection of antisense, sense, missense oligonucleotides and saline respectively, in the substantia nigra compacta of each rat via the cannula, followed by MPTP (30 mg/kg) injection. Behavior of the rats was observed and immunohistochemistry was carried out to check the expression of DAT and apoptosis of dopamine cell. RESULTS: The expression of DAT (positive unit, PU) in the substantia nigra compacta in rats was 6.65+/- 1.67 in the antisense ODN group, 12.41+/- 2.46 in saline group, 11.45+/- 1.17 in sense ODN group, and 10.35+/- 2.89 in missense ODN group. The expression of DAT was lower in the antisense ODN group than that of the other three groups (P< 0.01). The rotation of the rats induced by apomorphine was slower than that of the other three groups(P< 0.05). The apoptotic cells (21.4+/- 5.6) in the antisense ODN group (200x ) were less than that of the other three groups (61.6+/- 19.7, 56.5+/ 16.3, 52.2+/- 12.5 respectively), (P< 0.01). CONCLUSION: The expression of DAT can be inhibited effectively by the antisense ODN, and the response of the rats to the MPTP was reduced upon DAT inhibition. PMID- 18393257 TI - [Polymorphisms of four microsatellite DNA markers from telomeric HLA I region (D6S1624, D6S258, M6S211 and D6S510) and their linkage disequilibrium with HLA-A in a southern Chinese Han population]. AB - OBJECTIVE: To explore the genetic polymorphisms of four microsatellite DNA markers from telomeric HLA I region (D6S1624, D6S258, M6S211 and D6S510) and their linkage disequilibrium with HLA-A in a southern Chinese Han population residing in Hunan province. METHODS: Fluorescent PCR/Size-sequencing was carried out to analyze the polymorphisms of D6S1624, D6S258, M6S211 and D6S510 loci, and polymerase chain reaction-sequence specific priming (PCR-SSP) technique was used for HLA-A typing. RESULTS: The genotypic distributions at the 5 loci were consistent with Hardy-Weinberg equilibrium (P> 0.05). The number of allelic variants for D6S1624, D6S258, M6S211 and D6S510 loci were 10, 10, 12 and 9, respectively. Each locus had several main alleles and the dominant alleles were D6S1624-*199, D6S258-*195, M6S211-*261 and D6S510-*186. All of the 4 microsatellite markers exhibited high heterozygosity values (0.7142-0.8316) and polymorphism information content values (0.6686-0.811). No global linkage disequilibrium (LD) was detected between D6S1624 and HLA-A (P= 0.2646), or between D6S258 and HLA-A (P= 0.3481). In contrast, very significant global LD was found between M6S211 and HLA-A (P< 0.0001), and between D6S510 and HLA-A(P< 0.0001). Subsequent analysis for haplotypes with an observed frequency of > or = 3% revealed that only 2 of the 10 D6S1624-HLA-A haplotypes and 3 of the 9 D6S258 HLA-A haplotypes displayed weak or moderate LD, while 7 out of the 8 M6S211-HLA-A haplotypes, 6 among the 7 D6S510-HLA-A haplotypes were in tight LD. CONCLUSION: Authors have characterized four microsatellite DNA markers, D6S1624, D6S258, M6S211 and D6S510 in a southern Chinese Han population. Findings shown here can be helpful for those studies mainly addressing the association between HLA I sub region and diseases. The data also provide basis for future study in forensics, HLA matching in clinical transplantation and anthropology. PMID- 18393258 TI - Associative versus dissociative binding of CO to 4d transition metal trimers: A density functional study. AB - Density functional calculations were performed to determine equilibrium geometrical structures, transition states and relative energies for M(3) clusters (M = Nb, Mo, Tc, Ru, Rh, Pd, Ag) reacting with CO, leading to proposed reaction pathways. For the Nb(3), Mo(3), and Tc(3) clusters, the lowest energy structure correlates to dissociated CO, with the C and O atoms bound on opposite sides of the metal triangle. For all other trimers, the lowest energy structures maintain the CO moiety. In the case of Pd(3) and Ag(3) the dissociated geometries lie higher in energy than the sum of the separated reactants. In most cases, several multiplicities were found to be similar in energy and for Mo(3)CO and Pd(3)CO singlet-triplet minimum energy crossing points were identified. In the case of Rh(3)CO, minimum energy crossing points for the doublet, quartet, and sextet reaction pathways were determined and compared. The electron densities of pertinent M(3)CO species were investigated using Natural Bond Order calculations. It was found that the effect of the metal trimer on the energy of the pure p-type pi* antibonding orbital of carbon monoxide directly correlates with the occurrence of CO dissociation. PMID- 18393259 TI - Five new bidesmoside triterpenoid saponins from Stauntonia chinensis. AB - Eleven triterpenoid saponins (1-11) were isolated from Stauntonia chinensis DC. (Lardizabalaceae), including five new compounds, yemuoside YM(21-25) (1-3, 6, 7) structures of which were elucidated by chemical methods and a combination of MS, 1D- and 2D- NMR experiments including DEPT, (1)H--(1)H COSY, HSQC, HMBC, TOCSY, and NOESY as 3-O-alpha-L-arabinopyranosyl-(1 --> 3)-[alpha-L-rhamnopyranosyl-(1 - > 2)-]alpha-L-arabinopyranosyl-akebonicacid-28-O-alpha-L-rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (1), 3-O-beta-D xylopyranosyl-(1 --> 3)-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L arabinopyranosyl-akebonic acid-28-O-alpha-L-rhamnopyranosyl-(1 --> 4)-beta-D glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (2), 3-O-beta-D glucopyranosyl-(1 --> 3)-alpha-L-arabinopyranosyl-akebonic acid-28-O-alpha-L rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (3), 3-O-alpha-L-arabinopyranosyl-(1 --> 3)-[alpha-L-rhamnopyranosyl-(1 --> 2)-]alpha-L-arabinopyranosyl-akebonic acid-28-O-beta-D-glucopyranosyl-(1 --> 6) beta-D-glucopyranosyl ester (6), 3-O-alpha-L-arabinopyranosyl-(1 --> 3)-[alpha-L arabinopyranosyl-(1 --> 2)-]alpha-L-arabinopyranosyl-akebonic acid-28-O-beta-D glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl ester (7). PMID- 18393260 TI - Nanowire-decorated microscale metallic electrodes. PMID- 18393261 TI - Stabilization of DNA multilayer films through oligonucleotide crosslinking. AB - Multilayer films and capsules synthesized from DNA are of interest because they are biodegradable, biocompatible, and the structure of the films can be finely controlled by base pairing of the nucleotides. As DNA films are held together through a balance between the attractive hydrogen bonding and the aromatic stacking of the base pairs and the electrostatic repulsion of the negatively charged phosphate backbones, the films can be subject to disintegration at low salt concentrations (<200 mM). Enhancement of the stability of the films is essential if they are to be used in bioapplications. Herein, we describe an approach to form DNA films and capsules that are stable under a variety of buffer conditions, including low salt concentrations (down to 25 mM NaCl). The films are assembled using a triblock oligonucleotide system, in which the two outer blocks facilitate the assembly of the film and the middle block can be used to stabilize the films by hybridizing oligonucleotide sequences that crosslink the films. Additionally, crosslinked DNA capsules are shown to exhibit significantly different shrinkage properties to those of noncrosslinked capsules, thus demonstrating further control over the capsule properties. These DNA capsules are envisaged to find applications as drug-delivery vehicles, in diagnostics, and as microreactors. PMID- 18393262 TI - Understanding the patterns and distribution of opioid analgesic dependence symptoms using a latent empirical approach. AB - Prevalence of extramedical opioid analgesic use in the US is rising, yet little is known about the nature and extent of problems of dependence related to the use of these drugs. This study uses Latent Class Analysis to empirically define classes of past-year extramedical opioid analgesic users based on observed clustering of DSM-IV defined clinical dependence features; multinomial logistic regression is used to describe differences across these groups. The 2002-2003 public data-files of the National Survey on Drug Use and Health were used to identify 7810 extramedical opioid analgesic users in the past-year. The best fitting four-class model identified classes that differed quantitatively and qualitatively, with 2% of the users in Class 4 (most severe) and 84% in Class 1 (least severe). Classes 2 and 3 had parallel symptom profiles, but those in Class 3 reported additional problems. Adolescents (12-17 year olds) were at higher odds of being in Class 3 versus older age groups; females were two times as likely to be in Classes 2 and 4, and those with mental health problems were at higher odds of belonging to the more severe classes. Differences by type of past year opioid users were also detected. This study sheds light on the classification and distribution of extramedical opioid analgesic dependence symptoms in the US general population, identifying subgroups that warrant immediate attention. PMID- 18393263 TI - The reliability and validity of the forensic Camberwell Assessment of Need (CANFOR): a needs assessment for forensic mental health service users. AB - No instrument exists that measures the individual needs of forensic mental health service users (FMHSUs). The aim of this study was therefore to develop a valid and reliable individual needs assessment instrument for FMHSUs that incorporated staff and service user views and measured met and unmet needs. The Camberwell Assessment of Need was used as a template to develop CANFOR. Consensual and content validity were investigated with 50 forensic mental health professionals and 60 FMHSUs. Both were found to be satisfactory. Concurrent validity was tested using the Global Assessment of Functioning and a five-point needs scale, and again was found to be satisfactory. Reliability studies were carried out with 77 service users and 65 staff in high and medium security psychiatric services in the UK. Inter-rater reliability, rating whether a need was present or not, was high for service users (0.991) and staff (0.998). Similarly high reliability was found for unmet needs (0.985 and 0.972, respectively). Test-retest reliability was found to be moderately high for service users (0.795) and staff (0.852) when ratings were made two weeks apart. Similar levels were found for ratings of unmet needs (0.813 and 0.699, respectively). The average interview time was 23 minutes. CANFOR has good validity and reliability, and is suitable for further testing with other service user groups. PMID- 18393264 TI - Personal exposure to mobile phone frequencies and well-being in adults: a cross sectional study based on dosimetry. AB - The use of mobile phone telecommunication has increased in recent years. In parallel, there is growing concern about possible adverse health effects of cellular phone networks. We used personal dosimetry to investigate the association between exposure to mobile phone frequencies and well-being in adults. A random population-based sample of 329 adults living in four different Bavarian towns was assembled for the study. Using a dosimeter (ESM-140 Maschek Electronics), we obtained an exposure profile over 24 h for three mobile phone frequency ranges (measurement interval 1 s, limit of determination 0.05 V/m). Exposure levels over waking hours were totalled and expressed as mean percentage of the International Commission on Non-Ionizing Radiation Protection (ICNIRP) reference level. Each participant reported acute symptoms in a day-long diary. Data on five groups of chronic symptoms and potential confounders were assessed during an interview. The overall exposure to high-frequency electromagnetic fields was markedly below the ICNIRP reference level. We did not find any statistically significant association between the exposure and chronic symptoms or between the exposure and acute symptoms. Larger studies using mobile phone dosimetry are warranted to confirm these findings. PMID- 18393266 TI - Photochromism of a 3D Cd(II) complex with two captured ligand isomers generated in situ from the same precursor. PMID- 18393265 TI - A chiral hypervalent iodine(III) reagent for enantioselective dearomatization of phenols. PMID- 18393267 TI - A viable hydrogen-storage system based on selective formic acid decomposition with a ruthenium catalyst. PMID- 18393268 TI - Conformationally restricted hydantoin-based peptidomimetics as inhibitors of caspase-3 with basic groups allowed at the S3 enzyme subsite. AB - By using a combination of molecular modeling, combinatorial chemistry, and biological essays, novel scaffold molecules for the inhibition of caspase-3 have been developed. These compounds have an overall attenuated negative charge and show similar IC(50) values for both recombinant and human endogenous caspase-3. This might provide the basis for a novel strategy for the discovery of potent and more druglike inhibitors of caspase-3. PMID- 18393270 TI - Carcinoid tumor of the appendix: ultrasound findings in early pregnancy. AB - Ultrasound examination of a woman in early pregnancy with right lower quadrant abdominal pain demonstrated an edematous appendix with amorphous fluid surrounding the appendix. At laparotomy, these findings were confirmed. On pathological evaluation following surgical removal of the appendix a rare case of carcinoid tumor of the appendix was diagnosed. This is the first description of the transvaginal ultrasound features of a carcinoid tumor of the appendix. PMID- 18393269 TI - Arrested oligodendrocyte lineage maturation in chronic perinatal white matter injury. AB - OBJECTIVE: Abnormal myelination is a major pathological sequela of chronic periventricular white matter injury in survivors of premature birth. We tested the hypothesis that myelination failure in chronic hypoxia-ischemia-induced periventricular white matter injury is related to persistent depletion of the oligodendrocyte (OL) precursor pool required to generate mature myelinating OLs. METHODS: A neonatal rat model of hypoxia-ischemia was used where acute degeneration of late OL progenitors (preOLs) occurs via a mostly caspase independent mechanism. The fate of OL lineage cells in chronic cerebral lesions was defined with OL lineage-specific markers. RESULTS: Acute caspase-3 independent preOL degeneration from hypoxia-ischemia was significantly augmented by delayed preOL death that was caspase-3-dependent. Degeneration of preOLs was offset by a robust regenerative response that resulted in a several-fold expansion in the pool of surviving preOLs in chronic lesions. However, these preOLs displayed persistent maturation arrest with failure to differentiate and generate myelin. When preOL-rich chronic lesions sustained recurrent hypoxia ischemia at a time in development when white matter is normally resistant to injury, an approximately 10-fold increase in caspase-dependent preOL degeneration occurred relative to lesions caused by a single episode of hypoxia-ischemia. INTERPRETATION: The mechanism of myelination failure in chronic white matter lesions is related to a combination of delayed preOL degeneration and preOL maturation arrest. The persistence of a susceptible population of preOLs renders chronic white matter lesions markedly more vulnerable to recurrent hypoxia ischemia. These data suggest that preOL maturation arrest may predispose to more severe white matter injury in preterm survivors that sustain recurrent hypoxia ischemia. PMID- 18393271 TI - Risk factors for readmission after initial diagnosis in children with acute lymphoblastic leukemia. AB - BACKGROUND: Specific hospital discharge criteria following the initial diagnosis of children with acute lymphoblastic leukemia (ALL) have not been reported. This retrospective cohort study was designed to identify risk factors for readmission during induction therapy, to assist with development of discharge guidelines. PROCEDURE: We reviewed the records of 142 consecutive children with newly diagnosed B-precursor ALL and found 129 eligible patients. Chi square, t-test, and multivariate logistic regression analysis were used to compare differences in absolute neutrophil count (ANC), NCI risk status, age, type of corticosteroid administered, and other potential risk factors for readmission during induction therapy. RESULTS: ANC at initial hospital discharge was the only significant predictor of readmission for fever during induction therapy (P = 0.006) by multivariate analysis. Specifically an ANC 200/mm(3), in a clinically stable patient, is associated with minimal risk of readmission during induction therapy following the initial diagnosis of ALL. PMID- 18393272 TI - Alternative splicing of Nav1.5: an electrophysiological comparison of 'neonatal' and 'adult' isoforms and critical involvement of a lysine residue. AB - In developmentally regulated D1:S3 splicing of Nav1.5, there are 31 nucleotide differences between the 5'-exon ('neonatal') and the 3'-exon ('adult') forms, resulting in 7 amino acid differences in D1:S3-S3/S4 linker. In particular, splicing replaces a conserved negative aspartate residue in the 'adult' with a positive lysine. Here, 'neonatal' and 'adult' Nav1.5 alpha-subunit splice variants were stably transfected into EBNA-293 cells and their electrophysiological properties investigated by whole-cell patch-clamp recording. Compared with the 'adult' isoform, the 'neonatal' channel exhibited (1) a depolarized threshold of activation and voltage at which the current peaked; (2) much slower kinetics of activation and inactivation; (3) 50% greater transient charge (Na(+)) influx; (4) a stronger voltage dependence of time to peak; and (5) a slower recovery from inactivation. Tetrodotoxin sensitivity and VGSCbeta1-4 mRNA expression levels did not change. The significance of the charge-reversing aspartate to lysine substitution was investigated by mutating the lysine in the 'neonatal' channel back to aspartate. In this 'neonatal K211D' mutant, the electrophysiological parameters studied strongly shifted back towards the 'adult', that is the lysine residue was primarily responsible for the electrophysiological effects of Nav1.5 D1:S3 splicing. Taken together, these data suggest that the charge reversal in 'neonatal' Nav1.5 would (1) modify the channel kinetics and (2) prolong the resultant current, allowing greater intracellular Na(+) influx. Developmental and pathophysiological consequences of such differences are discussed. PMID- 18393273 TI - Altered expression of endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 in transformed non-lymphoid human tissues. AB - The endoplasmic reticulum (ER) aminopeptidases ERAP1 and ERAP2 contribute to generate HLA class I binding peptides. Recently, we have shown that the expression of these enzymes is high and coordinated (with each other and with HLA class I molecules) in immortalized B cells, but variable and imbalanced in human tumour cell lines of various non-lymphoid lineages. Herein, this issue was investigated in vivo by testing ERAP1 and ERAP2 expression in normal non-lymphoid tissues and their malignant counterparts. ERAP1 and ERAP2 were detected exclusively in the epithelial cells of over half of the tested normal tissues. Four ERAP1/ERAP2 phenotypes (+/+, -/-, +/- and -/+) were detected, and the presence of either or both enzymes was not necessarily associated with HLA class I expression. In more than 160 neoplastic lesions, the expression of either or both aminopeptidases was retained, lost (most frequently, particularly ERAP1) or acquired as compared to the normal counterparts, depending on the tumour histotype. The double-negative (-/-) phenotype was the most frequent, and significantly (P = 0.013) associated with a lack of detectable HLA class I antigens. In selected neoplastic lesions, ERAP1 and ERAP2 were also tested for their enzymatic (peptide-trimming) activities. Expression and function were found to correlate, indicating that immunohistochemistry detects active enzymes in vivo. Thus, dissociation in the expression of ERAP1, ERAP2 and HLA class I may already be present in some normal tissues, but malignant transformation causes additional losses, gains and imbalances in specific tumour histotypes, and these alter the peptide-trimming ability of tumour cells in vivo. PMID- 18393274 TI - Structure and function of 2:1 DNA polymerase.DNA complexes. AB - DNA polymerases are required for DNA replication and DNA repair in all of the living organisms. Different DNA polymerases are responsible different stages of DNA metabolism, and many of them are multifunctional enzymes. It was generally assumed that the different reactions are catalyzed by the same enzyme molecule. In addition to 1:1 DNA polymerase.DNA complex reported by crystallization studies, 2:1 and higher order DNA polymerase.DNA complexes have been identified in solution studies by various biochemical and biophysical approaches. Further, abundant evidences for the DNA polymerase-DNA interactions in several DNA polymerases suggested that the 2:1 complex represents the more active form. This review describes the current status of this emerging subject and explores their potential in vitro and in vivo functional significance, particularly for the 2:1 complexes of mammalian DNA polymerase beta (Pol beta), the Klenow fragment of E. coli DNA polymerase I (KF), and T4 DNA polymerase. PMID- 18393275 TI - Domperidone interferes with conditioned disgust reactions but not taste avoidance evoked by a LiCl-paired taste in infant rats. AB - Rats exhibit taste avoidance and conditioned disgust reactions when stimulated with a tastant paired with lithium chloride (LiCl). Lithium-mediated activation of chemoreceptor nuclei at the brainstem appears to determine the acquisition of conditioned taste aversion (CTA) in adult rodents. Domperidone (DOM), an anti emetic drug that does not cross the blood-brain barrier, was employed to analyze mechanisms underlying LiCl-mediated CTA in infant rats. On postnatal day 13 animals were given DOM followed by a pairing between intraoral saccharin and LiCl. Saccharin consumption at testing was lower in lithium-treated pups than in controls. DOM did not interfere with this LiCl-mediated taste avoidance but significantly decreased LiCl-mediated disgust reactions (head-shaking and wall climbing). Activation of the emetic system of the brainstem does not seem necessary for the acquisition of LiCl-mediated conditioned taste avoidance. Yet, these centers seem to be involved in the palatability shift resulting from taste LiCl pairings. These results indicate an early dissociation between conditioned disgust reactions and conditioned taste avoidance. PMID- 18393277 TI - Isolation and the timing of mammary gland development, gonadarche, and ovarian senescence: implications for mammary tumor burden. AB - In this study of Norway rats, we hypothesized that lifelong psychosocial experiences, social isolation or group living, trigger different developmental trajectories in the ovarian system, contributing to predisease pathways for spontaneous mammary tumors. Epidemiological studies indicate that early puberty and delayed menopause are risk factors for breast cancer. To that end, we took a cross-sectional, prospective approach and examined the ovarian system at two developmental points, young adulthood and middle age. We assessed ovarian function at both points, as well as mammary gland development at puberty and mammary tumor burden in middle age. Social isolation dissociated two components of puberty; it accelerated maturation of ovarian function while it simultaneously delayed mammary tissue development thereby increasing the exposure of developing breast parenchyma to high levels of estrogen. By mid-life, socially isolated rats had greater tumor burden despite having entered estropause prematurely, demonstrating that isolation did not increase tumorigenesis by prolonging ovarian function. These findings are discussed in the context of facultative lifespan strategies for rats born at different times of year and those living in isolation or in a large burrow community. PMID- 18393276 TI - Inflammatory responses of endothelial cells experiencing reduction in flow after conditioning by shear stress. AB - Exposure of endothelial cells (EC) to shear stress reduces their response to tumour necrosis factor-alpha (TNF). We tested how shear-conditioned EC responded to reduction in flow, either by spontaneously binding leukocytes, or by increasing sensitivity to TNF. Human umbilical vein EC were exposed to shear stress of 2.0 Pa (20 dyn/cm(2)) for 24 h. Shear was then reduced to stasis (30 sec perfusion each hour to exchange medium) or 0.003 Pa (creeping flow). At chosen times, neutrophils were perfused over the EC at 0.1 Pa (effective reperfusion). EC developed an ability to capture flowing neutrophils that lasted from 1 to 3 h after flow reduction, which was reduced by antibody against P selectin or pre-treatment of EC with an inhibitor of NADPH-oxidase. Adhesion of neutrophils to TNF-treated EC was greatly suppressed by shear-conditioning, remained suppressed immediately after cessation of flow and then took 48 h to approach the level in static cultures. Interestingly, the response to TNF remained suppressed in cultures switched to creeping flow. Gene array analysis confirmed that differently recovered cells had separate phenotypes. Thus, an acute response of EC to reduction in shear may contribute to leukocyte recruitment, along with hypoxia, in ischaemia and reperfusion. Prolonged cessation of flow may increase the sensitivity of EC to inflammatory stimuli, but this effect may be suppressed by residual flow. PMID- 18393278 TI - Parasympathetic and sympathetic responses to the strange situation in infants and mothers from avoidant and securely attached dyads. AB - Vagal reactivity and salivary alpha-amylase (sAA) were assessed in infants (M age = 13.55 months) and their mothers during the Strange Situation Paradigm (SSP) to investigate differences in physiological responses in a sample of insecure avoidant and securely-attached dyads (N = 132). Infants classified as insecure avoidant had significantly higher vagal withdrawal during the SSP and higher sAA overall, suggesting that the avoidant attachment pattern is associated with a greater allostatic load. During separation episodes of the SSP, all mothers showed significant vagal withdrawal, suggesting greater attempts at regulation. During the last reunion, typically the most stressful episode for infants, mothers of secure infants showed greater vagal withdrawal than mothers of insecure-avoidant infants, suggesting greater attempts by these mothers at interactive repair. Results for mothers and infants supported the allostatic load theory. PMID- 18393279 TI - A prospective study of diurnal cortisol responses to the social experience of school transition in four-year-old children: anticipation, exposure, and adaptation. AB - This study examined psychosocial influences on hypothalamic-pituitary-adrenal axis activity in 105 4-year-old children transitioning to primary school. Measuring before, during, and after school transition over a period of up to 12 months, salivary cortisol was assessed on awakening and early evening. Parents reported child temperament and teachers recorded adaptive behavior. Whilst cortisol at awakening and early evening increased from baseline to school transition, effects were not significant. A significant decrease occurred between transition and follow-up and from baseline to follow-up for both awakening and evening cortisol. Poorer effortful control was associated with high morning and steeper diurnal slope of cortisol at transition whilst surgency/extroversion was associated individually with greater morning and evening cortisol at transition and adaptation. Greater increase in internalizing social isolation during the first 6 months of school in more surgent/extrovert children predicted higher morning and evening cortisol at follow-up. This study is the first to explore these adaptive relationships over a 12-month period and supports social isolation over time as a key element in developmental endocrine activation. PMID- 18393280 TI - No stable arm preference during the pre-reaching period: a comparison of right and left hand kinematics with and without a toy present. AB - Adult hand preference emerges from complex developmental changes in arm and hand use during childhood. Recent reports have highlighted the importance of understanding arm and hand use during the first year of life including the period before reach onset. This longitudinal study tested the hypothesis that significant right-left differences exist in pre-reaching arm movements. We examined right and left hand kinematics from 13 healthy infants during trials with and without a toy present from 8 weeks of age through the week of reach onset. Significant right-left differences were found, however there was no clear pattern within a condition or across conditions. Without a toy present, the right hand moved faster, yet ended further from midline, and displayed more movements during the Late phase compared to other phases. With a toy present, the right hand moved longer lengths, yet ended movements further away from the toy. When left and right hand kinematics were combined, previous findings of right hand kinematics alone were supported. Although infants begin adapting their pre reaching kinematics many weeks before reach onset, we did not find evidence of a systematic right--left difference before reach onset in movements with or without a toy present. Our results, coupled with other reports, suggest hand asymmetries begin to emerge over the year following reach onset amid developmental changes both within the infant, and the physical and social environment. PMID- 18393281 TI - Social play in crested and Japanese macaques: testing the covariation hypothesis. AB - Most studies about social play have focused on its function in aiding socialization. It is not known whether play patterns vary by species according to specific adult social relationships. To test whether there is covariation between the form of play interactions and adult relationships, we studied social play in Japanese macaques (Macaca fuscata) and crested macaques (Macaca nigra). Comparative studies have shown that whereas low levels of tolerance and strict hierarchies characterize Japanese macaques, crested macaques display greater tolerance and relaxed dominance. The results showed that the former prefer less risky games, with minimal physical contact, ease of escape and a small number of play partners. The play bouts of the latter species involve closer contact and more partners. We conclude that play patterns covary with specific social style. Play patterns could have been selected for to introduce youngsters to their future social environment. PMID- 18393282 TI - Could adults be used to improve social skills of young horses, Equus caballus? AB - We investigated the effects of the introduction of foreign adults on the behavior of young horses. First, we observed the behavior of 1- and 2-year-old domestic horses housed in same-age and same-sex groups (a standard housing system, but different from a natural situation). Then, two same-sex adults were introduced into each experimental group. Observations made before, during and after an introduction indicated that young horses reared in homogeneous groups of young had different behaviors compared to other domestic horses reared under more socially natural conditions. After the introduction of adults, young horses expressed new behaviors, preferential social associations emerged, positive social behavior increased and agonistic interactions decreased. These results have important implications both for understanding the influence that adults may have on the behavior of young horses, and in terms of husbandry, indicating the importance of keeping young horses with adults, although further studies are still necessary. PMID- 18393284 TI - Effects of early deprivation and maternal separation on pup-directed behavior and HPA axis measures in the juvenile female rat. AB - Juvenile female rats show maternal-like behavior toward pups. The purpose of the following experiment was to investigate whether the HPA axis, through the use of early separation manipulations that alter HPA functioning in rats, plays a role in the juvenile response to foster pups. Female rats were early deprived or maternally separated for 5 hours daily from PND 2 to 14 and compared to animal facility-reared rats. Deprivation or separation increased CRH-R1 IR in the juvenile PVN, but had no other effects on other HPA measures or on maternal behavior. Pup-exposure during the juvenile period blunted corticosterone levels after acute and repeated pup-exposures when compared to exposure to novelty and conspecifics respectively. Repeated exposures to pups also increased CRH-R1 IR relative to isolation during the juvenile period. Overall, the data suggest that although pup-exposure affects corticosterone levels, the HPA axis does not relate to juvenile maternal behavior in the present experiments. PMID- 18393285 TI - Rats selectively bred for low levels of 50 kHz ultrasonic vocalizations exhibit alterations in early social motivation. AB - In rats, the rates of 50 kHz ultrasonic vocalizations (USVs) can be used as a selective breeding phenotype and variations in this phenotype can be an indicator of affective states. The 50 kHz USV is elicited by rewarding stimuli (e.g., food, sexual behavior) and therefore can express a positive affective state. Conversely, the 22 kHz USV is elicited by aversive stimuli (e.g., presence of a predator, social defeat) indicating a negative affective state. In the present study, we tested the effect of selectively breeding for 50 kHz USVs on a variety of maternal social/emotional behaviors in young rat pups (PND 10-12). These measures consisted of an assessment of isolation calls and conditioned odor preference paradigm. Results indicate that animals selected for low levels of 50 kHz USVs show the greatest alterations in social behaviors compared to the control animals. The low line animals had an increase in isolation calls tested during place preference conditioning and a decrease in 50 kHz ultrasonic calls in all conditions. These same low line animals failed to show a typical preference for a maternally-associated odor during the place preference test. The different social behaviors of the high line animals did not consistently vary from those of the control group. These results have important implications for the study of genetic and epigenetic mechanisms underlying emotional states, and possibly contribute to the research underlying the emotional changes in developmental disorders such as autistic spectrum disorder by providing a novel animal model that displays communication deficits that are interdependent with significant social behavioral impairments. PMID- 18393286 TI - Effects of early weaning on anxiety and prefrontal cortical and hippocampal myelination in male and female Wistar rats. AB - We investigated developmental changes in myelin formation in the prefrontal cortex and the hippocampus, and behavioral effects of early weaning in Wistar rats. Early-weaned rats showed decreased numbers of open-arm entries in an elevated plus-maze in both sexes at 4 weeks old; this effect persisted in males, but ceased in females after this age. Expression of myelin basic protein (MBP) showed both age-dependent increases and sex differences; 4-week-old males exhibited higher MBP levels in the hippocampus, whereas 7-week-old males showed lower MBP levels in the prefrontal cortex compared to females of the same age. There was a tendency for group differences from weaning for the 21.5-kDa isoform in the prefrontal cortex. Although these results suggest that male rats are more vulnerable than females to early-weaning effects on anxiety-related behaviors, further detailed analysis is needed to clarify the functional relationship between myelination and anxiety-related behaviors. PMID- 18393287 TI - Apolipoprotein B-dependent hepatitis C virus secretion is inhibited by the grapefruit flavonoid naringenin. AB - Hepatitis C virus (HCV) infects over 3% of the world population and is the leading cause of chronic liver disease worldwide. HCV has long been known to associate with circulating lipoproteins, and its interactions with the cholesterol and lipid pathways have been recently described. In this work, we demonstrate that HCV is actively secreted by infected cells through a Golgi dependent mechanism while bound to very low density lipoprotein (vLDL). Silencing apolipoprotein B (ApoB) messenger RNA in infected cells causes a 70% reduction in the secretion of both ApoB-100 and HCV. More importantly, we demonstrate that the grapefruit flavonoid naringenin, previously shown to inhibit vLDL secretion both in vivo and in vitro, inhibits the microsomal triglyceride transfer protein activity as well as the transcription of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase and acyl-coenzyme A:cholesterol acyltransferase 2 in infected cells. Stimulation with naringenin reduces HCV secretion in infected cells by 80%. Moreover, we find that naringenin is effective at concentrations that are an order of magnitude below the toxic threshold in primary human hepatocytes and in mice. CONCLUSION: These results suggest a novel therapeutic approach for the treatment of HCV infection. PMID- 18393288 TI - Acetylsalicylic acid inhibits hepatitis C virus RNA and protein expression through cyclooxygenase 2 signaling pathways. AB - It has been reported that salicylates (sodium salicylate and aspirin) inhibit the replication of flaviviruses, such as Japanese encephalitis virus and dengue virus. Therefore, we considered it important to test whether acetylsalicylic acid (ASA) had anti-hepatitis C virus (HCV) activity. To this end, we examined the effects of ASA on viral replication and protein expression, using an HCV subgenomic replicon cell culture system. We incubated Huh7 replicon cells with 2 8 mM ASA for different times and measured HCV-RNA and protein levels by northern blot, real-time polymerase chain reaction, and western analysis, respectively. We found that ASA had a suppressive effect on HCV-RNA and protein levels (nearly 58%). ASA-dependent inhibition of HCV expression was not mediated by the 5' internal ribosome entry site or 3'-untranslated regions, as determined by transfection assays using bicistronic constructs containing these regulatory regions. However, we found that HCV-induced cyclooxygenase 2 (COX-2) messenger RNA and protein levels and activity and these effects were down-regulated by ASA, possibly by a nuclear factor kappa B-independent mechanism. We also observed that the ASA-dependent inhibition of viral replication was due in part to inhibition of COX-2 and activation of p38 and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1/2 (MEK1/2) mitogen-activated protein kinases (MAPKs). Inhibition of these kinases by SB203580 and U0126, respectively, and by short interfering RNA silencing of p38 and MEK1 MAPK prevented the antiviral effect of ASA. Taken together, our findings suggest that the anti-HCV effect of ASA in the Huh7 replicon cells is due to its inhibitory effect on COX-2 expression, which is mediated in part by the activation of MEK1/2/p38 MAPK. CONCLUSION: These findings suggest the possibility that ASA could be an excellent adjuvant in the treatment of chronic HCV infection. PMID- 18393289 TI - Noninvasive assessment of hepatic lipid composition: Advancing understanding and management of fatty liver disorders. AB - Nonalcoholic fatty liver is frequently observed in obese individuals, yet the factors that predict its development and progression to liver disease are poorly understood. We proposed that proton magnetic resonance spectroscopy ((1)H-MRS) might allow noninvasive assessment of hepatic lipid composition. Lipid saturation (SI) and polyunsaturation (PUI) indices measured by (1)H-MRS were in agreement with those expected in oils of known composition. Hepatic triglyceride concentration (HTGC) and composition were then measured in healthy lean (LEAN) men, obese men with normal HTGC (OB), and obese men with hepatic steatosis (OB+HS). The effect of marked changes in dietary fat consumption on hepatic lipids were also compared in lean men after 67 hours of a normal mixed (NM) diet versus a low-carbohydrate, high-saturated-fat (LCHF) diet. SI was significantly higher in OB+HS (0.970 +/- 0.004) and OB (0.944 +/- 0.008) versus LEAN (0.818 +/- 0.025) (P < 0.01 for both). PUI was significantly lower in OB+HS (0.003 +/- 0.001) and OB (0.022 +/- 0.005) versus LEAN (0.120 +/- 0.021) (P < 0.01), and significantly lower in OB+HS versus OB (P < 0.05). LCHF diet did not alter HTGC, SI, or PUI (P > 0.05). The (1)H-MRS method provides for rapid, qualitative assessment of lipid composition. Application of this technique in the liver produces results that are consistent with biopsy-based approaches demonstrating that relative hepatic lipid saturation increases and polyunsaturation decreases with obesity. Obesity-related hepatic steatosis is characterized by further depletion of polyunsaturated hepatic lipids. CONCLUSION: This readily available and noninvasive approach should promote further study into interactions between hepatic and whole-body lipid metabolism and help to elucidate the pathogenesis of disorders characterized by lipid accumulation within the liver. PMID- 18393291 TI - Two patients with atypical interstitial deletions of 8p23.1: mapping of phenotypical traits. AB - Chromosomal 8p23 deletion syndrome is recognized as a malformation syndrome with clinical symptoms of facial anomalies, microcephaly, mental retardation, and congenital heart defects. The responsible gene for the heart defects in this syndrome has been identified as GATA4 on 8p23.1. Two patients with interstitial deletions of 8p23.1 were investigated; one patient showed moderate developmental delay and Ebstein anomaly, and the other showed mild delay and typical atrioventricular septum defect. The precise deletion sizes, 17 and 2.9 Mb, were determined by FISH analyses using BAC clones as probes. The latter deletion was the smallest deletion including GATA4 in the previously reported patients, and the critical regions and genes for clinical manifestation of 8p23 deletion syndrome, including facial anomalies, microcephaly, behavioral abnormality, and developmental delay, were discussed. PMID- 18393290 TI - Interactions of pluronic block copolymers on P-gp efflux activity: experience with HIV-1 protease inhibitors. AB - The objective was to examine the influence of Pluronic block-copolymers on the interaction between the drug efflux transporter, P-glycoprotein and HIV-1 protease inhibitors (PIs). The ATPase assay determined the effect of various Pluronics on PI-stimulated P-gp ATPase activity. Cellular accumulation studies were conducted using MDCKII and LLC-PK1 cells transfected with human MDR1 to assess Pluronic modulation of PI efflux. Pluronic P85 inhibited both basal and nelfinavir-stimulated P-gp ATPase activity, while Pluronic F127 had no effect. In cell accumulation studies, Pluronic P85 restored the accumulation of nelfinavir in MDCKII-MDR1 cells while Pluronic F127 and F88 had no effect. Pluronic P85 increased saquinavir accumulation in wild-type and MDR1-transfected cells in both the MDCKII and LLC-PK1 cell models, suggesting inhibition of multiple transporters, including MRPs. In conclusion, this study provides evidence that a block-copolymer, Pluronic P85, effectively inhibits the interaction of P-gp with nelfinavir and saquinavir. These data indicate that effective inhibition of HIV-1 PI efflux by Pluronic P85 may influence the distribution of antiretroviral agents to sites protected by efflux mechanisms, such as the blood-brain barrier, and possibly increase the brain exposure of these drugs resulting in suppression of viral replication and reduction in the incidence of drug resistant mutants. PMID- 18393292 TI - TIS21 negatively regulates hepatocarcinogenesis by disruption of cyclin B1 Forkhead box M1 regulation loop. AB - A functional and biochemical interaction of TIS21(/BTG2/PC3) with Forkhead box M1 (FoxM1), essential transcription factor for hepatocyte regeneration and a master regulator of mitotic gene expression, was explored. Growth of hepatocellular carcinoma (HCC), developed by a single injection of diethylnitrosamine (DEN), was the same in both the TIS21(+/+) and TIS21(-/-) mice until 6 months, whereas it was significantly higher in the TIS21(-/-) mice at 9 months. Expression of TIS21 was significantly lower in both human and murine HCCs than in the surrounding tissues. Forced expression of TIS21 impaired growth, proliferation, and tumorigenic potential of Huh7 cells. At the mechanistic level, TIS21 inhibited FoxM1 phosphorylation, a required modification for its activation, by reducing cyclin B1-cdk1 activity, examined by in vitro kinase assay and FoxM1 mutant analyses. These observations were further confirmed in vivo by the reciprocal control of TIS21 expression and FoxM1 phosphorylation in the diethylnitrosamine induced HCCs and TIS21(-/-) mouse embryonic fibroblast (MEF), in addition to increased expression of cyclin B1 and cdk1 activity. CONCLUSION: TIS21 negatively regulated hepatocarcinogenesis in part by disruption of the FoxM1-cyclin B1 regulatory loop, thereby inhibiting proliferation of transformed cells developed in mouse and human livers. PMID- 18393293 TI - Clinicopathological study on cholangiolocellular carcinoma suggesting hepatic progenitor cell origin. AB - Cholangiolocellular carcinoma (CLC), a subtype of cholangiocellular carcinoma (CC), is thought to originate from the ductules/canals of Hering, where hepatic progenitor cells (HPCs) are located. We investigated the clinicopathological features of 30 CLCs and their relationship to HPCs. We evaluated the expression of hepatocytic markers (hepatocyte paraffin-1, canalicular polyclonal carcinoembryonic antigen, and CD10), biliary/HPC markers (keratin [K]7, K19, and neural cell adhesion molecule), the adenosine triphosphate binding cassette transporters: multidrug resistance protein 1, multidrug resistance-associated protein (MRP)1, MRP3, and breast cancer resistance protein, using immunohistochemistry and electron microscopy. In addition, gene expression profiling of CLC was performed and compared with the profile of hepatocellular carcinoma (HCC) with or without HPC features (K19 expression). In surrounding nontumoral tissue, K7-positive and K19-positive HPCs/ductular reaction were observed. More than 90% of the tumor was composed of CLC areas that showed small monotonous and/or anastomosing glands, strongly positive for K7 and K19. Especially at the tumor boundary, all cases showed a HCC-like trabecular area characterized by canalicular CD10/polyclonal carcinoembryonic antigen expression, and submembranous K7 expression, similar to intermediate hepatocytes. K7 positive/K19-positive HPCs were also seen. Out of 30 cases, 19 showed papillary and/or clear glandular formation with mucin production, representing CC areas. These three different areas showed transitional zones with each other. We observed an increased expression of MRP1, MRP3, and breast cancer resistance protein in the tumor. Electron microscopy findings in HCC-like trabecular areas confirmed the presence of HPCs and intermediate hepatocytes. HPC markers, K7, K19, prominin-1, receptor for stem cell factor c-kit, octamer-4 transcription factor, and leukemia inhibitory factor were upregulated (P < 0.05), while albumin was downregulated in CLC (P = 0.007) toward K19-negative HCCs. Comparison of CLC with K19-positive HCCs indicated a high homology. CONCLUSION: All these findings highly suggest a progenitor cell origin of CLC. PMID- 18393294 TI - Development of enteric and vagal innervation of the zebrafish (Danio rerio) gut. AB - The autonomic nervous system develops following migration and differentiation of precursor cells originating in the neural crest. Using immunohistochemistry on intact zebrafish embryos and larvae we followed the development of the intrinsic enteric and extrinsic vagal innervation of the gut. At 3 days postfertilization (dpf), enteric nerve cell bodies and fibers were seen mainly in the middle and distal intestine, while the innervation of the proximal intestine was scarcer. The number of fibers and cell bodies gradually increased, although a large intraindividual variation was seen in the timing (but not the order) of development. At 11-13 dpf most of the proximal intestine received a similar degree of innervation as the rest of the gut. The main intestinal branches of the vagus were similarly often already well developed at 3 dpf, entering the gut at the transition between the proximal and middle intestine and projecting posteriorly along the length of the gut. Subsequently, fibers branching off the vagus innervated all regions of the gut. The presence of several putative enteric neurotransmitters was suggested by using markers for neurokinin A (NKA), pituitary adenylate cyclase-activating polypeptide (PACAP), vasoactive intestinal polypeptide (VIP), nitric oxide, serotonin (5-hydroxytryptamine, 5-HT), and calcitonin gene-related peptide (CGRP). The present results corroborate the belief that the enteric innervation is well developed before the onset of feeding (normally occurring around 5-6 dpf). Further, the more detailed picture of how development proceeds at stages previously not examined suggests a correlation between increasing innervation and more regular and elaborated motility patterns. PMID- 18393295 TI - Nucleus paragigantocellularis afferents in male and female rats: organization, gonadal steroid receptor expression, and activation during sexual behavior. AB - The supraspinal regulation of genital reflexes is poorly understood. The brainstem nucleus paragigantocellularis (nPGi) of rats is a well-established source of tonic inhibition of genital reflexes. However, the organization, gonadal steroid receptor expression, and activity of nPGi afferents during sex have not been fully characterized in male and female rats. To delineate the anatomical and physiological organization of nPGi afferents, the retrograde tracer Fluoro-Gold (FG) was injected into the nPGi of sexually experienced male and female rats. Animals engaged in sexual behavior 1 hour before sacrifice. Cells containing FG, estrogen receptor-alpha (ER(alpha)), androgen receptor (AR), and the immediate-early gene product Fos were identified immunocytochemically. Retrograde labeling from the nPGi was prominent in the bed nucleus of the stria terminalis, paraventricular nucleus (PVN), posterior hypothalamus, precommissural nucleus, deep mesencephalic nucleus, and periaqueductal gray (PAG) of both sexes. Sex differences were observed in the caudal medial preoptic area (MPO), with significantly more FG+ cells observed in males, and in the PAG and inferior colliculus, where significantly more FG+ cells were observed in females. The majority of regions that contained FG+ cells also contained ER(alpha) or AR, indicating sensitivity to gonadal steroids. The proportions of FG+ cells that co localized with sex-induced Fos was high in the PVN of both sexes and high in the MPO of males but low in the PAG of both sexes despite the large number of PAG nPGi output neurons and Fos+ cells in both sexes. The characterization of these afferents will lead to a further understanding of the neural regulation of genital reflexes. PMID- 18393296 TI - Nondestructive optical determination of fiber organization in intact myocardial wall. AB - Mapping the myocardial fiber organization is important for assessing the electrical and mechanical properties of normal and diseased hearts. Current methods to determine the fiber organization have several limitations: histological sectioning mechanically distorts the tissue and is labor-intensive, while diffusion tensor imaging has low spatial resolution and requires expensive MRI scanners. Here, we utilized optical clearing, a fluorescent dye, and confocal microscopy to create three-dimensional reconstructions of the myocardial fiber organization of guinea pig and mouse hearts. We have optimized the staining and clearing procedure to allow for the nondestructive imaging of whole hearts with a thickness up to 3.5 mm. Myocardial fibers could clearly be identified at all depths in all preparations. We determined the change of fiber orientation across strips of guinea pig left ventricular wall. Our study confirms the qualitative result that there is a steady counterclockwise fiber rotation across the ventricular wall. Quantitatively, we found a total fiber rotation of 105.7+/-14.9 degrees (mean+/-standard error of the mean); this value lies within the range reported by previous studies. These results show that optical clearing, in combination with a fluorescent dye and confocal microscopy, is a practical and accurate method for determining myocardial fiber organization. PMID- 18393297 TI - Changes in metformin pharmacokinetics after intravenous and oral administration to rats with short-term and long-term diabetes induced by streptozotocin. AB - It has been reported that metformin was primarily metabolized via hepatic CYP2C11, 2D1, and 3A1/2 in rats, and the expression and mRNA levels of hepatic CYP2C11 and 3A1 decreased and increased, respectively, whereas the expression of CYP2D1 was not changed in rat model of diabetes induced by streptozotocin (DMIS). Also minimizing the toxic effects of streptozotocin by carrying out experiments 4 5 weeks after streptozotocin injection has been reported. Thus, the pharmacokinetics of metformin was evaluated in rat model of DMIS at the 7th and the 29th days after streptozotocin injection. After intravenous administration of metformin (100 mg/kg) to rat model of DMIS, the CL(R) became significantly faster (46.9% and 77.8% increase for the 7th and the 29th days, respectively; due to urine flow rate-dependent timed-interval renal clearance of the drug) and CL(NR) became significantly slower (28.0% and 34.3% decrease, respectively; due to decreased hepatic CYP2C11) than in their respective controls. After oral administration of metformin (100 mg/kg) to rat model of DMIS, the AUC became significantly smaller (18.6% and 33.7% decrease for the 7th and the 29th days, respectively) than in their respective controls. The CL(NR) of metformin were comparable between two rat models of DMIS. PMID- 18393298 TI - Pharmacokinetics of lofexidine hydrochloride in healthy volunteers. AB - The objective of this study was to characterize the clinical pharmacokinetic profile of lofexidine after oral delivery. A single dose, cross-over study and a multidose study using healthy volunteers were conducted for that purpose. In the single dose study the average time to maximum concentration was observed at approximately 3 h for the single doses tested (1.2 mg dose and 2.0 mg). Area under the curve from time zero to infinity (AUC(0-infinity)) demonstrated a degree of dose proportionality with a 1.72-fold increase as the dose increased by a factor of 1.67. Elimination rates and terminal half-lives were comparable between dose levels. The average elimination rates for the 1.2 mg and the 2.0 mg doses were 0.063 and 0.065 h(-1), respectively. In the multidose study, the average maximum concentration observed after the first dose of 0.4 mg was 433 ng/L and ranged from 338 to 586 ng/L. This was slightly lower in proportion to the maximum concentration observed in the single dose study where C(max) was 1755 ng/L at the 1.2 mg dose (normalized to 585 ng/L for 0.4 mg dose) and for the 2.0 mg dose (normalized to 559 ng/L for 0.4 mg dose). The average time to maximum concentration (T(max)) was 3.33 h which is comparable to values observed in the single dose study. The pharmacokinetic data indicate that lofexidine has a consistent profile. Steady state seems to be reached after 2 days on lofexidine, which is consistent with the lofexidine elimination half-life of approximately 11 h. Evaluation of the T(max), elimination rate, and terminal half-life are consistent across all dose levels studied, suggesting that changing the dose does not affect the absorption or elimination rates of lofexidine HCl. Thus, although preliminary due to the limited number of subjects, these findings are the first to document lofexidine clinical pharmacokinetic parameters in healthy volunteers using a highly sensitive liquid chromatography tandem mass spectrometric analysis. PMID- 18393299 TI - Solid state characterization and crystal structure from X-ray powder diffraction of two polymorphic forms of ranitidine base. AB - Ranitidine hydrochloride (RAN-HCl), a known anti-ulcer drug, is the product of reaction between HCl and ranitidine base (RAN-B). RAN-HCl has been extensively studied; however this is not the case of the RAN-B. The solid state characterization of RAN-B polymorphs has been carried out using different analytical techniques (microscopy, thermal analysis, Fourier transform infrared spectrometry in the attenuated total reflection mode, (13)C-CPMAS-NMR spectroscopy and X-ray powder diffraction). The crystal structures of RAN-B form I and form II have been determined using conventional X-ray powder diffraction in combination with simulated annealing and whole profile pattern matching, and refined using rigid-body Rietveld refinement. RAN-B form I is a monoclinic polymorph with cell parameters: a = 7.317(2), b = 9.021(2), c = 25.098(6) A, beta = 95.690(1) degrees and space group P2(1)/c. The form II is orthorhombic: a = 31.252(4), b = 13.052(2), c = 8.0892(11) A with space group Pbca. In RAN-B polymorphs, the nitro group is involved in a strong intramolecular hydrogen bond responsible for the existence of a Z configuration in the enamine portion of the molecules. A tail to tail packing motif can be denoted via intermolecular hydrogen bonds. The crystal structures of RAN-B forms are compared to those of RAN-HCl polymorphs. RAN-B polymorphs are monotropic polymorphic pairs. PMID- 18393300 TI - Stability of the Hsp90 inhibitor 17AAG hydroquinone and prevention of metal catalyzed oxidation. AB - 17-(allylamino)-17-demethoxygeldanamycin (17AAG) is a benzoquinone ansamycin Hsp90 inhibitor which has promising anticancer activity in vitro, in animal models and in clinical trials. 17AAG has poor water-solubility which is a potential problem for clinical formulation. The hydroquinone derivative of 17AAG, 17AAG hydroquinone (17AAGH(2)), is considerably more water soluble and since we previously demonstrated that 17AAGH(2) was a more potent Hsp90 inhibitor than its parent quinone, it is a good candidate for clinical use and is currently in clinical trials. However, 17AAGH(2) can be oxidized back to 17AAG under aerobic conditions so we tested the relative stability of 17AAGH(2) and the effect of different metal ions and metal chelators on the oxidation of 17AAGH(2). We found that copper could accelerate 17AAGH(2) oxidation while copper chelators such as D penicillamine could inhibit oxidation. Human serum albumin (HA) has copper binding ability and we found that HA diminished the rate of 17AAGH(2) oxidation. Although we found that 17AAG could associate with HA, no association was observed between 17AAGH(2) and HA. In summary, our data demonstrates that copper chelators can prevent 17AAGH(2) oxidation and suggests that HA prevents 17AAGH(2) oxidation via a copper chelation mechanism. Agents that prevent oxidation may be useful in clinical formulations of 17AAGH(2.) PMID- 18393301 TI - Temporal-spatial activation of apoptosis and epithelial injury in murine experimental biliary atresia. AB - Biliary atresia is a fibro-inflammatory cholangiopathy that obstructs the extrahepatic bile ducts in young infants. Although the pathogenesis of the disease is undefined, studies in livers from affected children and neonatal mice with experimental biliary atresia have shown increased expression of proapoptosis molecules. Therefore, we hypothesized that apoptosis is a significant mechanism of injury to duct epithelium. To test this hypothesis, we quantified apoptosis using terminal transferase dUTP nick end labeling and active caspase-3 staining in livers and extrahepatic bile ducts from Balb/c mice infected with Rhesus rotavirus (RRV) within 24 hours of birth. RRV induced a significant increase in labeled cells in the portal tracts and in epithelial and subepithelial compartments of extrahepatic bile ducts, with onset within 3 days and peaks at 5 10 days. Exploring mechanisms of injury, we found increased biliary expression of caspases 1 and 4 and of interferon-gamma (IFNgamma)-related and tumor necrosis factor-alpha (TNFalpha)-related genes. Using a cholangiocyte cell line, we found that neither IFNgamma nor TNFalpha alone affected cell viability; however, simultaneous exposure to IFNgamma and TNFalpha activated caspase-3 and decreased cell viability. Inhibition of caspase activity blocked apoptosis and restored viability to cultured cholangiocytes. In vivo, administration of the caspase inhibitor IDN-8050 decreased apoptosis in the duct epithelium and the extent of epithelial injury after RRV challenge. CONCLUSION: The biliary epithelium undergoes early activation of apoptosis in a mouse model of biliary atresia. The synergistic role of IFNgamma and TNFalpha in activating caspase-3 in cholangiocytes and the decreased apoptosis following pharmacologic inhibition of caspases support a prominent role for apoptosis in the pathogenesis of experimental biliary atresia. PMID- 18393302 TI - Angiofil-mediated visualization of the vascular system by microcomputed tomography: a feasibility study. AB - Visualization of the vascular systems of organs or of small animals is important for an assessment of basic physiological conditions, especially in studies that involve genetically manipulated mice. For a detailed morphological analysis of the vascular tree, it is necessary to demonstrate the system in its entirety. In this study, we present a new lipophilic contrast agent, Angiofil, for performing postmortem microangiography by using microcomputed tomography. The new contrast agent was tested in 10 wild-type mice. Imaging of the vascular system revealed vessels down to the caliber of capillaries, and the digital three-dimensional data obtained from the scans allowed for virtual cutting, amplification, and scaling without destroying the sample. By use of computer software, parameters such as vessel length and caliber could be quantified and remapped by color coding onto the surface of the vascular system. The liquid Angiofil is easy to handle and highly radio-opaque. Because of its lipophilic abilities, it is retained intravascularly, hence it facilitates virtual vessel segmentation, and yields an enduring signal which is advantageous during repetitive investigations, or if samples need to be transported from the site of preparation to the place of actual analysis, respectively. These characteristics make Angiofil a promising novel contrast agent; when combined with microcomputed tomography, it has the potential to turn into a powerful method for rapid vascular phenotyping. PMID- 18393303 TI - AFM characterization of rabbit spermatozoa. AB - Atomic force microscopy (AFM) has been applied for determining the topological and structural features of rabbit spermatozoa. Fresh ejaculated spermatozoa were adsorbed passively onto a silicon slide or by motility from suspension onto a poly(L-lysine)-coated glass coverslip and then imaged in air and in buffer saline, respectively. AFM images clearly highlighted many details of spermatozoa head, neck, and tail. Distinct features were observed in the plasmatic membrane of spermatozoa. In particular, head topography easily recognized the acrosome, equatorial segment, equatorial subsegment, and postacrosome regions. Moreover, AFM images revealed the presence of double belt of invaginations around the spermatozoa head, at the boundary between equatorial subsegment and postacrosome regions. All together, the collected AFM images clearly defined a detailed map of spermatozoa morphology while giving some hints on the internal structure. PMID- 18393304 TI - Transneuronal retrograde viral labeling in the brain stem and hypothalamus is more intense from the left than from the right adrenal gland. AB - Previous studies using the viral transneuronal tracing technique demonstrated central autonomic circuits involved in the innervation of the adrenal gland. Since increasing number of data indicate laterality in the neuroendocrine system, we aimed to investigate whether the supraspinal innervation of the adrenal gland exhibits asymmetry or not. The central circuitry involved in the innervation of the left and the right adrenal gland was studied in individual rats by dual transneuronal tracing using isogenic recombinant strains (Ba-DupGreen and Ba Duplac expressing lacZ) of Bartha strain of pseudorabies virus. Viral infection of brain nuclei (dorsal vagal nucleus, nucleus of the solitary tract, caudal raphe nuclei, A5 cell group, hypothalamic paraventricular nucleus) from the left adrenal was more severe than that from the right organ. Dual-infected neurons were present both in the brain stem and in the hypothalamus. The results indicate a predominance in the supraspinal innervation of the left adrenal gland, and that each adrenal gland is innervated both by side-specific neurons and by neurons that project to both organs. PMID- 18393305 TI - Nodal and lefty signaling regulates the growth of pancreatic cells. AB - Nodal and its antagonist, Lefty, are important mediators specifying the laterality of the organs during embryogenesis. Nodal signals through activin receptors in the presence of its co-receptor, Cripto. In the present study, we investigated the possible roles of Nodal and Lefty signaling during islet development and regeneration. We found that both Nodal and Lefty are expressed in the pancreas during embryogenesis and islet regeneration. In vitro studies demonstrated that Nodal inhibits, whereas Lefty enhances, the proliferation of a pancreatic cell line. In addition, we showed that Lefty-1 activates MAPK and Akt phosphorylation in these cells. In vivo blockade of endogenous Lefty using neutralizing Lefty-1 monoclonal antibody results in a significantly decreased proliferation of duct epithelial cells during islet regeneration. This is the first study to decipher the expression and function of Nodal and Lefty in pancreatic growth. Importantly, our results highlight a novel function of Nodal Lefty signaling in the regulation of expansion of pancreatic cells. PMID- 18393307 TI - Mice with an anterior cleft of the palate survive neonatal lethality. AB - Many genes are known to function in a region-specific manner in the developing secondary palate. We have previously shown that Shox2-deficient embryos die at mid-gestation stage and develop an anterior clefting phenotype. Here, we show that mice carrying a conditional inactivation of Shox2 in the palatal mesenchyme survive the embryonic and neonatal lethality, but develop a wasting syndrome. Phenotypic analyses indicate a delayed closure of the secondary palate at the anterior end, leading to a failed fusion of the primary and secondary palates. Consistent with a role proposed for Shox2 in skeletogenesis, Shox2 inactivation causes a significantly reduced bone formation in the hard palate, probably due to a down-regulation of Runx2 and Osterix. We conclude that the secondary palatal shelves are capable of fusion with each other, but fail to fuse with the primary palate in a developmentally delayed manner. Mice carrying an anterior cleft can survive neonatal lethality. PMID- 18393306 TI - Retinoic acid induces prostatic bud formation. AB - Formation of prostatic buds from the urogenital sinus (UGS) to initiate prostate development requires localized action of several morphogenetic factors. This report reveals all-trans-retinoic acid (RA) to be a powerful inducer of mouse prostatic budding that is associated with reciprocal changes in expression of two regulators of budding: sonic hedgehog (Shh) and bone morphogenetic protein 4 (Bmp4). Localization of retinoid signaling and expression of RA synthesis, metabolism, and receptor genes in the UGS on embryonic days 14.5-17.5 implicate RA in the mechanism of bud initiation. In UGS organ culture, RA increased prostatic budding, increased Shh expression, and decreased Bmp4. Prostatic budding was stimulated in the absence of RA by recombinant SHH, by blocking BMP4 signaling with NOGGIN, or by combined treatment with SHH and NOGGIN in UGS organ culture media. These observations suggest that reciprocal changes in hedgehog and BMP signaling by RA may regulate bud initiation. PMID- 18393308 TI - Bipolar, anastral spindle development in artificially activated sea urchin eggs. AB - The mitotic apparatus of the early sea urchin embryo is the archetype example of a centrosome-dominated, large aster spindle organized by means of the centriole of the fertilizing sperm. In this study, we tested the hypothesis that artificially activated sea urchin eggs possess the capacity to assemble the anastral, bipolar spindles present in many acentrosomal systems. Control fertilized Lytechinus pictus embryos and ammonia-activated eggs were immunolabeled for tubulin, centrosomal material, the spindle pole structuring protein NuMA and the mitotic kinesins MKLP1/Kinesin-6, Eg5/Kinesin-5, and KinI/Kinesin-13. Confocal imaging showed that a subset of ammonia-activated eggs contained bipolar "mini-spindles" that were anastral; displayed metaphase and anaphase-like stages; labeled for centrosomal material, NuMA, and the three mitotic kinesins; and were observed in living eggs using polarization optics. These results suggest that spindle structural and motor proteins have the ability to organize bipolar, anastral spindles in sea urchin eggs activated in the absence of the paternal centriole. PMID- 18393309 TI - Morphallactic regeneration as revealed by region-specific gene expression in the digestive tract of Enchytraeus japonensis (Oligochaeta, Annelida). AB - Enchytraeus japonensis is a small oligochaete, which primarily reproduces asexually by fragmentation and regeneration. For precise analysis of the pattern formation during regeneration, we isolated three region-specific genes (EjTuba, mino, and horu) expressed in the digestive tract. In growing worms, the expression of EjTuba in the head and mino in the trunk region just posterior to the head were observed in defined body segments, while the expression areas of EjTuba in the trunk and horu were proportional to the total number of body segments. In the regeneration process, expression of these genes disappeared once and recovered to their original pattern by day 7. In abnormal regeneration such as a bipolar head, mino was still expressed in the region next to both the normal and the ectopic heads. These results suggest that there is morphallactic as well as epimorphic or inductive regulation of the body patterning during regeneration of E. japonensis. PMID- 18393310 TI - Assembly of primary cilia. AB - Primary cilia are microtubule-based, hair-like sensory organelles present on the surface of most growth-arrested cells in our body. Recent research has demonstrated a crucial role for primary cilia in regulating vertebrate developmental pathways and tissue homeostasis, and defects in genes involved in primary cilia assembly or function have been associated with a panoply of disorders and diseases, including polycystic kidney disease, left-right asymmetry defects, hydrocephalus, and Bardet Biedl Syndrome. Here we provide an up-to-date review focused on the molecular mechanisms involved in the assembly of primary cilia in vertebrate cells. We present an overview of the early stages of the cilia assembly process, as well as a description of the intraflagellar transport (IFT) system. IFT is a highly conserved process required for assembly of almost all eukaryotic cilia and flagella, and much of our current knowledge about IFT is based on studies performed in Chlamydomonas and Caenorhabditis elegans. Therefore, our review of the IFT literature includes studies performed in these two model organisms. The role of several non-IFT proteins (e.g., centrosomal proteins) in the ciliary assembly process is also discussed. PMID- 18393312 TI - The fate of Pluronic F-68 in chondrocytes and CHO cells. AB - The surfactant Pluronic F-68 (PF-68) is widely used in large-scale mammalian cell culture to protect cells from shear stress that arises from agitation and gas sparging. Several studies suggested that PF-68 is incorporated into the cell plasma membrane and could enter the cells, but without providing any direct evidence. The current study has examined this question for two cell types, one of pharmaceutical interest (CHO cells) and the other of biomedical interest (chondrocytes or cartilage cells). A fluorescent derivative of PF-68 was synthesized to detect and localize internalized Pluronic with culture time. PF-68 uptake by the cells was quantified and characterized. We clearly demonstrate that PF-68 enters the cells, and possibly accumulates in the endocytic pathway. CHO cells showed an average uptake of 11.7 +/- 6.7 (SEM) microg PF-68/10(6) cells while the uptake of chondrocytes was 56.0 +/- 10.9 (SEM) microg PF-68/10(6) cells, independently of the initial PF-68 concentration (between 0.01 and 0.2%, w/v) and of cell concentration (from 1 x 10(6) to 4 x 10(6) cells/mL). These uptake values were identical for both static and agitated culture conditions. Finally, we found that CHO cells are able to eliminate intracellular fluorescent PF-68 but chondrocytes are not. These results show that the uptake of PF-68 by the cells can severely affect PF-68 concentration in the culture medium and thus shear protection effect. PMID- 18393311 TI - FGF signaling segregates biliary cell-lineage from chick hepatoblasts cooperatively with BMP4 and ECM components in vitro. AB - Intrahepatic bile ducts (IHBDs) are indispensable for transporting bile secreted from hepatocytes to the hepatic duct. The biliary epithelial cells (BECs) of the IHBD arise from bipotent hepatoblasts around the portal vein, suggesting the portal mesenchyme is essential for their development. However, except for Notch or Activin/TGF-beta signaling molecules, it is not known which molecules regulate IHBD development. Here, we found that FGF receptors and BMP4 are specifically expressed in the developing IHBD and the hepatic mesenchyme, respectively. Using a mesenchyme-free culture of liver bud, we showed that bFGF and FGF7 induce the hepatoblasts to differentiate into BECs, and that BMP4 enhances bFGF-induced BEC differentiation. The extracellular matrix (ECM) components in the hepatic mesenchyme induced BEC differentiation. Forced expression of a constitutively active form of the FGF receptor partially induced BEC differentiation markers in vivo. These data strongly suggest that bFGF and FGF7 promote BEC differentiation cooperatively with BMP4 and ECMs in vivo. PMID- 18393313 TI - Distinctive microbial ecology and biokinetics of autotrophic ammonia and nitrite oxidation in a partial nitrification bioreactor. AB - Biological nitrogen removal (BNR) based on partial nitrification and denitrification via nitrite is a cost-effective alternate to conventional nitrification and denitrification (via nitrate). The goal of this study was to investigate the microbial ecology, biokinetics, and stability of partial nitrification. Stable long-term partial nitrification resulting in 82.1 +/- 17.2% ammonia oxidation, primarily to nitrite (77.3 +/- 19.5% of the ammonia oxidized) was achieved in a lab-scale bioreactor by operation at a pH, dissolved oxygen and solids retention time of 7.5 +/- 0.1, 1.54 +/- 0.87 mg O(2)/L, and 3.0 days, respectively. Bioreactor ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) populations were most closely related to Nitrosomonas europaea and Nitrobacter spp., respectively. The AOB population fraction varied in the range 61 +/- 45% and was much higher than the NOB fraction, 0.71 +/- 1.1%. Using direct measures of bacterial concentrations in conjunction with independent activity measures and mass balances, the maximum specific growth rate (micro(max)), specific decay (b) and observed biomass yield coefficients (Y(obs)) for AOB were 1.08 +/- 1.03 day(-1), 0.32 +/- 0.34 day(-1), and 0.15 +/- 0.06 mg biomass COD/mg N oxidized, respectively. Corresponding micro(max), b, and Y(obs) values for NOB were 2.6 +/- 2.05 day(-1), 1.7 +/- 1.9 day(-1), and 0.04 +/- 0.02 mg biomass COD/mg N oxidized, respectively. The results of this study demonstrate that the highly selective partial nitrification operating conditions enriched for a narrow diversity of rapidly growing AOB and NOB populations unlike conventional BNR reactors, which host a broader diversity of nitrifying bacteria. Further, direct measures of microbial abundance enabled not only elucidation of mixed community microbial ecology but also estimation of key engineering parameters describing bioreactor systems supporting these communities. PMID- 18393314 TI - Application of high-performance tangential flow filtration (HPTFF) to the purification of a human pharmaceutical antibody fragment expressed in Escherichia coli. AB - High-performance tangential flow filtration (HPTFF) is shown to successfully enable concentration, purification and formulation in a single unit operation. This is illustrated with feedstreams comprising recombinant proteins expressed in Escherichia coli (E. coli). Using positively charged cellulosic membranes of 100 kDa molecular weight cut-off and operating under a selected range of buffer pH and ionic strength at a filtrate flux of 100 L m(-2) h(-1), a 10-fold removal of E. coli host cell proteins (HCP) was obtained with an overall process yield of 98%. The HPTFF performance was shown to be robust and reproducible. In addition, the novel charged membrane was regenerated and re-used seven times without loss of selectivity or throughput. When compared with a conventional purification scheme, the proposed process results in the elimination of one chromatographic step, a 12% yield improvement and a significant reduction in purification cost of goods. PMID- 18393315 TI - Activation of immobilized lipase in non-aqueous systems by hydrophobic poly-DL tryptophan tethers. AB - Many industrially important reactions use immobilized enzymes in non-aqueous, organic systems, particularly for the production of chiral compounds such as pharmaceutical precursors. The addition of a spacer molecule ("tether") between a supporting surface and enzyme often substantially improves the activity and stability of enzymes in aqueous solution. Most "long" linkers (e.g., polyethylene oxide derivatives) are relatively hydrophilic, improving the solubility of the linker-enzyme conjugate in polar environments, but this provides little benefit in non-polar environments such as organic solvents. We present a novel method for the covalent immobilization of enzymes on solid surfaces using a long, hydrophobic polytryptophan tether. Candida antarctica lipase B (CALB) was covalently immobilized on non-porous, functionalized 1-microm silica microspheres, with and without an intervening hydrophobic poly-DL-tryptophan tether (n approximately 78). The polytryptophan-tethered enzyme exhibited 35 times greater esterification of n-propanol with lauric acid in the organic phase and five times the hydrolytic activity against p-nitrophenol palmitate, compared to the activity of the same enzyme immobilized without tethers. In addition, the hydrophobic tethers caused the silica microspheres to disperse more readily in the organic phase, while the surface-immobilized control treatment was less lipophilic and quickly settled out of the organic phase when the suspensions were not vigorously mixed. PMID- 18393316 TI - Cytochrome P450 2E1 contributes to ethanol-induced fatty liver in mice. AB - Cytochrome P450 2E1 (CYP2E1) is suggested to play a role in alcoholic liver disease, which includes alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. In this study, we investigated whether CYP2E1 plays a role in experimental alcoholic fatty liver in an oral ethanol-feeding model. After 4 weeks of ethanol feeding, macrovesicular fat accumulation and accumulation of triglyceride in liver were observed in wild-type mice but not in CYP2E1-knockout mice. In contrast, free fatty acids (FFAs) were increased in CYP2E1-knockout mice but not in wild-type mice. CYP2E1 was induced by ethanol in wild-type mice, and oxidative stress induced by ethanol was higher in wild-type mice than in CYP2E1 knockout mice. Peroxisome proliferator-activated receptor alpha (PPARalpha), a regulator of fatty acid oxidation, was up-regulated in CYP2E1-knockout mice fed ethanol but not in wild-type mice. A PPARalpha target gene, acyl CoA oxidase, was decreased by ethanol in wild-type but not in CYP2E1-knockout mice. Chlormethiazole, an inhibitor of CYP2E1, lowered macrovesicular fat accumulation, inhibited oxidative stress, and up-regulated PPARalpha protein level in wild-type mice fed ethanol. The introduction of CYP2E1 to CYP2E1-knockout mice via an adenovirus restored macrovesicular fat accumulation. These results indicate that CYP2E1 contributes to experimental alcoholic fatty liver in this model and suggest that CYP2E1-derived oxidative stress may inhibit oxidation of fatty acids by preventing up-regulation of PPARalpha by ethanol, resulting in fatty liver. PMID- 18393317 TI - Hepatocytes can induce death of contacted cells via perforin-dependent mechanism. AB - The liver displays unique immunological properties including the ability to remove aberrant cells and pathogens and to induce peripheral immunotolerance. We have previously demonstrated that hepatocytes can cause cell death by a CD95 ligand-mediated mechanism. Here, we provide evidence that hepatocytes can kill other cells via a perforin-dependent pathway. Using cultured woodchuck hepatocytes and human liver cells as well as freshly isolated woodchuck, mouse, and human hepatocytes, we show that hepatocyte-mediated death of CD95-deficient target cells requires microtubule polymerization, a feature of the granule exocytosis-mediated cytotoxicity. Neutralizing anti-perforin antibodies and short hairpin RNA directed against perforin messenger RNA confirmed the involvement of perforin in hepatocyte-mediated cell killing. CONCLUSION: This study shows that hepatocytes express biologically competent perforin capable of killing susceptible cells and emphasizes the role of hepatocytes as cytotoxic effectors. This also is the first demonstration of perforin in a non-lymphoid cell type. PMID- 18393318 TI - Suppressor of cytokine signaling 1 protects mice against concanavalin A-induced hepatitis by inhibiting apoptosis. AB - Acute liver failure is associated with significant mortality. However, the underlying pathophysiological mechanism is not yet fully understood. Suppressor of cytokine signaling-1 (SOCS1), which is a negative-feedback molecule for cytokine signaling, has been shown to be rapidly induced during liver injury. Here, using liver-specific SOCS1-conditional-knockout mice, we demonstrated that SOCS1 deletion in hepatocytes enhanced concanavalin A (ConA)-induced hepatitis, which has been shown to be dependent on activated T and natural killer T (NKT) cells. Although serum cytokine level and NKT cell activation were similar in wild type (WT) and SOCS1-deficient mice after ConA treatment, proapoptotic signals, including signal transducers and activators of transcription 1 (STAT1) and Jun terminal kinase (JNK) activation, were enhanced in SOCS1-deficient livers compared with those in WT livers. SOCS1-deficient hepatocytes had higher expression of Fas antigen and were more sensitive to anti-Fas antibody-induced apoptosis than were WT hepatocytes. Furthermore, SOCS1-deficient hepatocytes were more sensitive to tumor necrosis factor (TNF)-alpha-induced JNK activation and apoptosis. These data indicate that SOCS1 is important to the prevention of hepatocyte apoptosis induced by Fas and TNF-alpha. In contrast, SOCS1 overexpression in the liver by adenoviral gene transfer prevented ConA-induced liver injury. CONCLUSION: These findings indicate that SOCS1 plays important negative roles in fulminant hepatitis and that forced expression of SOCS1 is therapeutic in preventing hepatitis. PMID- 18393320 TI - Loss of orphan receptor small heterodimer partner sensitizes mice to liver injury from obstructive cholestasis. AB - The orphan nuclear hormone receptor small heterodimer partner (SHP) regulates the expression of several genes involved in bile acid homeostasis in the liver. Because bile acid toxicity is a major source of liver injury in cholestatic disease, we explored the role of SHP in liver damage induced by common bile duct ligation (BDL). Shp(-/-) mice show increased sensitivity in this model of acute obstructive cholestasis, with greater numbers of bile infarcts and higher mortality than wild-type C57BL/6 mice. This increased sensitivity could not be accounted for by differences in expression of bile acid homeostatic genes 2 or 5 days after BDL. Instead, higher basal expression of such genes, including the key biosynthetic enzyme cholesterol 7alpha hydroxylase (Cyp7A1) and the bile salt export pump, is associated with both an increase in bile flow prior to BDL and an increase in acute liver damage at only 1.5 hours after BDL in Shp(-/-) mice, as shown by bile infarcts. At 3 hours, Cyp7A1 expression still remained elevated in Shp(-/-) with respect to wild-type mice, and the hepatic and serum bile acid levels and total hepatobiliary bile acid pool were significantly increased. The increased sensitivity of mice lacking SHP contrasts with the decreased sensitivity of mice lacking the farnesoid X receptor (FXR; nuclear receptor subfamily 1, group H, member 4) to BDL, which has been associated with decreased intraductal pressure and fewer bile infarcts. CONCLUSION: We propose that differences in acute responses to BDL, particularly the early formation of bile infarcts, are a primary determinant of the differences in longer term sensitivity of the Fxr(-/-) and Shp(-/-) mice to acute obstructive cholestasis. PMID- 18393321 TI - Hepatocyte-specific inhibitor-of-kappaB-kinase deletion triggers the innate immune response and promotes earlier cell proliferation during liver regeneration. AB - Nuclear factor kappaB (NF-kappaB) is one of the main transcription factors involved in liver regeneration after partial hepatectomy (PH). It is activated upon IkappaB phosphorylation by the IkappaB kinase (IKK) complex comprising inhibitor of kappaB kinase 1 (IKK1), inhibitor of kappaB kinase 2 (IKK2), and nuclear factor-B essential modifier (NEMO). We studied the impact of hepatocyte specific IKK2 deletion during liver regeneration. A 70% PH was performed on IKK2(f/f) (wild-type) and IKK2DeltaLPCmice (hepatocyte-specific IKK2 knockout mice). PH in IKK2DeltaLPC compared with IKK2(f/f) mice resulted in weaker and delayed NF-kappaB activation in hepatocytes, while nonparenchymal liver cells showed earlier NF-kappaB activation and higher tumor necrosis factor expression. Additionally, these animals showed increased and earlier serum amyloid A and chemotactic cytokine L-1 levels followed by enhanced polymorphonuclear cell recruitment to the liver. These results correlated with earlier Jun kinase activity, c-myc expression, and matrix metalloproteinase-9 activity, suggesting earlier priming in IKK2DeltaLPC mice after PH. These data preceded a more rapid cell cycle progression and earlier hepatocyte proliferation as evidenced through cyclin and 5-bromo-2-deoxyuridine analysis. Interestingly, despite faster G(1)/S progression, IKK2DeltaLPC mice exhibited an enduring mitosis phase, because mitotic bodies were still observed at later stages after PH. CONCLUSION: We demonstrate that PH in IKK2DeltaLPC mice triggers a more rapid and pronounced inflammatory response in nonparenchymal liver cells, which triggers earlier hepatocyte proliferation. PMID- 18393319 TI - Recombinant factor VIIa for variceal bleeding in patients with advanced cirrhosis: A randomized, controlled trial. AB - A beneficial effect of recombinant activated factor VII (rFVIIa) in Child-Pugh class B and C patients with cirrhosis who have variceal bleeding has been suggested. This randomized controlled trial assessed the efficacy and safety of rFVIIa in patients with advanced cirrhosis and active variceal bleeding. At 31 hospitals in an emergency setting, 256 patients (Child-Pugh > 8; Child-Pugh B = 26%, C = 74%) were randomized equally to: placebo; 600 microg/kg rFVIIa (200 + 4x 100 microg/kg); or 300 microg/kg rFVIIa (200 + 100 microg/kg). Dosing was intravenous at 0, 2, 8, 14, and 20 hours after endoscopy, in addition to standard vasoactive, prophylactic antibiotic, and endoscopic treatment. The primary composite endpoint consisted of failure to control 24-hour bleeding, or failure to prevent rebleeding or death at day 5. Secondary endpoints included adverse events and 42-day mortality. Baseline characteristics were comparable between groups. Administration of rFVIIa had no significant effect on the composite endpoint compared with placebo (P = 0.37). There was no significant difference in 5-day mortality between groups; however, 42-day mortality was significantly lower with 600 microg/kg rFVIIa compared with placebo (odds ratio 0.31, 95% confidence interval = 0.13-0.74), and bleeding-related deaths were reduced from 12% (placebo) to 2% (600 microg/kg). A marked heterogeneity in the failure rate in all treatment groups was observed across participating centers. Adverse events, including overall thromboembolic events, were comparable between groups. CONCLUSION: Treatment with rFVIIa had no significant effect on the primary composite endpoint compared with placebo. Therefore, decision on the use of this hemostatic agent in acute variceal bleeding should be carefully considered, because results of this study do not support the routine use of rFVIIa in this setting. Adverse events were comparable across groups. PMID- 18393323 TI - Cirrhosis and liver failure in nonalcoholic fatty liver disease: Molehill or mountain? PMID- 18393322 TI - Vascular biology and pathobiology of the liver: Report of a single-topic symposium. AB - Portal hypertension and its complications account for the majority of morbidity and mortality that occurs in patients with cirrhosis. In addition to portal hypertension, a number of other vascular syndromes are also of great importance, especially the ischemia-reperfusion (IR) injury. With the identification of major vascular defects that could account for many of the clinical sequelae of these syndromes, the liver vasculature field has now integrated very closely with the broader vascular biology discipline. In that spirit, the Henry and Lillian Stratton Basic Research Single Topic Conference was held on the topic of Vascular Biology and Pathobiology of the Liver. The course took place approximately 10 years after the first American Association for the Study of Liver Disease (AASLD) sponsored conference on this topic that occurred in Reston, Virginia. The conference initiated with an introduction to basic vascular cell signaling and then explored vascular biology specifically as it relates to liver cells. Subsequently, specific disease syndromes were discussed in more detail including portal hypertension and IR injury. Finally, clinical and translational sessions focused on emerging therapies and technologies to treat vascular diseases of the liver. PMID- 18393324 TI - Automated FISH analysis using dual-fusion and break-apart probes on paraffin embedded tissue sections. AB - Detecting balanced translocations using tissue sections plays an important diagnostic role in cases of hematological malignancies. Manual scoring is often problematic due to truncation and overlapping of nuclei. Reports have described automated analysis using primarily tile sampling. The aim of this study was to investigate an automated fluorescent in situ hybridization analysis method using grid sampling on tissue sections, and compare the performance of dual-fusion (DF) and break-apart (BA) probes in this setting. Ten follicular, 10 mantle cell lymphoma, and 10 translocation-negative samples were used to set the threshold of false positivity using IGH/CCND1, IGH/BCL-2 DF, and IGH BA probes. The cut-off distances of red and green signals to define fusion signals were 0.5, 1.0, and 1.2 mum for the IGH/CCND1, IGH/BCL-2 DF, and IGH BA probes, respectively. The mean false positivity of grid units was 5.3, 11.4, and 28.1%, respectively. Ten to 14 additional samples analyzed blindly and were correctly classified using each probe. Discriminating positive and negative samples using automated analysis and grid sampling was possible with each probe, although different definitions of fusion signals were required due to the different physical distances between the DNA probes. Using the DF probes resulted in lower false positivity, which was less affected by signal numbers per grid units. PMID- 18393325 TI - Improved survival time: what can survival cure models tell us about population based survival improvements in late-stage colorectal, ovarian, and testicular cancer? AB - BACKGROUND: The objective of the current study was to investigate the long-term impact of treatment advances on the survival of patients with late-stage ovarian, colorectal (American Joint Committee on Cancer stage III, men), and testicular cancers by estimating the increase in the percentage cured from their disease and the change in survival time of uncured patients. METHODS: Cause-specific survival data from 1973 to 2000 were obtained from the Surveillance, Epidemiology, and End Results Program. Survival cure models were fit and were used to estimate the gain in life expectancy (GLE) attributed to an increase in the fraction of cured patients and to prolonged survival among noncured patients. RESULTS: Treatment improvement for ovarian cancer resulted in a total GLE of 2 years, and 80% of that GLE was because of an extension of survival time in uncured patients (from 0.9 years to 2.1 years) rather than an increased cure fraction (from 12% to 14%). In contrast, the cure rate rose from 29% to 47% for colorectal cancer, representing 82% of a 2.8-year GLE, and from 23% to 81% for testicular cancer, representing 100% of a 24-year GLE. CONCLUSIONS: The current results suggested that treatment benefits for testicular and colorectal cancer in men with late stage disease primarily are the result of increases in cure fraction, whereas survival gains for ovarian cancer occur despite persisting disease. Cure models, in combination with population-level data, provide insight into how treatment advances are changing survival and ultimately impacting mortality. Survival patterns reflect the underlying biology of response to cancer treatment and suggest promising directions for future research. PMID- 18393326 TI - Androgen deprivation falls as orchiectomy rates rise after changes in reimbursement in the U.S. Medicare population. AB - BACKGROUND: Expenditures related to the use of medical androgen deprivation led in part to the Medicare Modernization Act (MMA) in 2003. This mandated a decline in reimbursement to 80% to 85% of the average wholesale price starting in 2004 followed by a more significant reduction in 2005 to 106% of the average sales price, which effectively reduced the reimbursement by approximately 50% of 2003 values. The authors hypothesized that these changes in reimbursement may affect the way practitioners administer these treatments. METHODS: The publicly available dataset Medicare Part B Extract Summary System was examined from 2001 to 2005 for trends in the number of allowed services and dollar amounts of allowed charges and payments. The reimbursable Medicare codes of J9217 (leuprolide acetate), J9202 (goserelin acetate), J9219 (leuprolide acetate implant), and J3315 (triptorelin pamoate) were examined for medical castration. The code for simple orchiectomy, 54520, was used for surgical castration. RESULTS: The use of medical castration increased from 2001 to 2003, whereas, over the same period, surgical castration decreased. Total allowed charges for medical castration peaked in 2003 at $1.23 billion. After the enactment of the MMA, surgical castration rates increased, and medical castration decreased. Total allowed charges for medical castration in 2005 dropped 65% from the 2003 peak. CONCLUSIONS: The use of medical androgen ablation decreased significantly with the decrease in reimbursement. The administration of either surgical or medical castration in the U.S. Medicare population appears to be tied closely to reimbursement in trend, but not always in magnitude. PMID- 18393327 TI - Charcot neuro-osteoarthropathy. AB - The classical neurotraumatic and neurotrophic theories for the pathogenesis of the acute Charcot neuro-osteoarthropathy (CN) in diabetes, do not address certain key features of the disease. These features include the facts that the condition usually affects just one side, that it is self-limiting, and that it is also very uncommon. Similarly, it is not known to what extent the condition may depend, as suggested by Jean-Martin Charcot, on pre-morbid osteopenia. Recent advances in understanding the mechanisms underlying the pathogenesis of osteopenia and osteoporosis and the central role of the RANKL/OPG signalling system have, however, suggested the possible involvement of other factors in the evolution of the disease. Specifically, it has been suggested that acute CN may be triggered in a susceptible individual by any event that leads to localized inflammation in the affected foot. This local inflammation leads to a vicious cycle in which there is increasing inflammation, increasing expression of RANKL, and increasing bone breakdown. The likely central role for the RANKL/OPG pathway suggests new possibilities for future treatments. PMID- 18393328 TI - The value of debridement and Vacuum-Assisted Closure (V.A.C.) Therapy in diabetic foot ulcers. AB - BACKGROUND: Treatment of diabetic foot ulcers includes a number of different regimes such as glycaemic control, re-vascularization, surgical, local wound treatment, offloading and other non-surgical treatments. Although considered the standard of care, the scientific evidence behind the various debridements used is scarce. This presentation will focus on debridement and V.A.C. Therapy, two treatments widely used in patients with diabetes and foot ulcers. METHODS: A review of existing literature on these treatments in diabetic foot ulcers, with focus on description of the various types of debridements used, the principles behind negative pressure wound therapy (NPWT) using the V.A.C. Therapy system and level of evidence. RESULTS: Five randomized controlled trials (RCT) of debridement were identified; three assessed the effectiveness of a hydrogel as a debridement method, one evaluated surgical debridement and one evaluated larval therapy. Pooling the three hydrogel RCTs suggested that hydrogels are significantly more effective than gauze or standard care in healing diabetic foot ulcers. Surgical debridement and larval therapy showed no significant benefit. Other debridement methods such as enzyme preparations or polysaccharide beads have not been evaluated in RCTs of people with diabetes. More than 300 articles have been published on negative pressure wound therapy, including several small RCTs and a larger multi-centre RCT of diabetic foot ulcers. Negative pressure wound therapy seems to be a safe and effective treatment for complex diabetic foot wounds, and could lead to a higher proportion of healed wounds, faster healing rates, and potentially fewer re-amputations than standard care. CONCLUSIONS: Although debridement of the ulcer is considered a prerequisite for healing of diabetic foot ulcers, the grade of evidence is quite low. This may be due to a lack of studies rather than lack of effect. Negative pressure wound therapy seems to be safe and effective in the treatment of some diabetic foot ulcers, although there is still only one well-performed trial that evaluates the effect. PMID- 18393329 TI - Interprofessional and transdisciplinary teamwork in health care. AB - The article focuses on the need for and the characteristics and positive consequences of interdisciplinary teamwork in health care. Interprofessional collaboration is an important element in total quality management. Factors that determine the success of team work are described, such as a management that promotes openness and an administrative organization that promotes interdisciplinary consultation. Other factors have to do with leadership, shared goals and values, meeting management and planning skills, communication, and also the (degree of) knowledge and the (quality of) perception of competences of other health care workers. The shared care plan is stressed as an important tool. In this, the joint planning of goals for intervention and care is essential. Health care workers with different professional knowledge and background have to harmonize their intervention plan according to the competences and goal settings of the other team members. The core of effective interprofessional teamwork is the presence of interprofessional competences such as these. A brief description of the components and performance criteria of the competence of interprofessional collaboration is given. PMID- 18393330 TI - Thermal unfolding of proteins probed by laser spray mass spectrometry. AB - The stability and conformational changes of cytochrome c (cyt c) at different temperatures and pH have been well examined so far by using various analytical methods. We have found that laser spray mass spectrometry enables much faster and more convenient monitoring of those changes of cyt c compared with other methods. The results correlated well with circular dichroism (CD) experiments under relatively acidic conditions, which destabilize the protein. Laser spray mass spectra of cyt c at various pH were obtained at different levels of laser power. Bimodal charge-state distributions of the protein were observed in laser spray mass spectra, indicating the two-state model of structural change; the lower charges correspond to the folded state, the higher charges to the unfolded state. Based on this result, the presumed denaturation curve of the protein was plotted as a function of laser power, and laser power by which 50% of the protein was assumed to be denatured, E50%, as obtained at each pH. We also examined the melting temperatures, Tm, of cyt c at various values of pH by using CD spectroscopy. The correlation coefficient between E50% and Tm for cyt c was 0.999, demonstrating an excellent correlation. Furthermore, laser spray analysis of ubiquitin, which is found to be more thermally stable than cyt c, gave a higher E50% than cyt c. These results indicate that laser spray mass spectrometry can be an extremely convenient method for probing thermal stabilities and dynamic conformational changes of proteins with subtle structural differences caused by slight changes in pH. PMID- 18393332 TI - A successful joint effort. PMID- 18393334 TI - Microcapillary electrophoresis chips utilizing controllable micro-lens structures and buried optical fibers for on-line optical detection. AB - In this study, a new design of a controllable micro-lens structure capable of the enhancement of LIF detection system has been demonstrated, which can be further integrated with buried optical fibers on a micro-CE chip for sample separation and detection. Two pneumatic side-chambers were placed between a micro-CE channel and an optical fiber channel. The intervals between the side-chamber and the microchannel were used to form two surfaces of the controllable micro-lens structure. Deformations of the two surfaces can be generated after pressurized index-matching fluid was injected into the pneumatic side-chambers. The side chambers can be deflected as a double convex lens to focus both the excitation light source and the fluorescent emission signal. The profile and the focal length of the micro-lens structure can be actively adjusted by applying different liquid pressures so that biosamples with a low concentration can be detected. Using low-cost polymeric materials such as polydimethylsiloxane, rapid and reliable fabrication techniques involving standard lithography and replication process was employed for the formation of the proposed chip device. Experimental results revealed the controllable micro-lens structure can be successfully deformed as a convex lens to focus the laser light source and the collected fluorescence signal can be enhanced accordingly. The power amplitude of excitation laser light can be enhanced by 5.4-fold. FITC dye and DNA markers were then utilized for micro-CE testing. The results indicated that the signal amplitude could be enhanced 2.5-fold when compared to the case without the activation of the micro-lens. According to the experimental results, the developed device has a great potential to be integrated with other microfluidic devices for further biomedical applications. PMID- 18393335 TI - A high-performance polycarbonate electrophoresis microchip with integrated three electrode system for end-channel amperometric detection. AB - A fully integrated polycarbonate (PC) microchip for CE with end-channel electrochemical detection operated in an amperometric mode (CE-ED) has been developed. The on-chip integrated three-electrode system consisted of a gold working electrode, an Ag/AgCl reference electrode and a platinum counter electrode, which was fabricated by photo-directed electroless plating combined with electroplating. The working electrode was positioned against the separation channel exit to reduce post-channel band broadening. The electrophoresis high voltage (HV) interference with the amperometric detection was assessed with respect to detection noise and potential shifts at various working-to-reference electrode spacing. It was observed that the electrophoresis HV interference caused by positioning the working electrode against the channel exit could be diminished by using an on-chip integrated reference electrode that was positioned in close proximity (100 microm) to the working electrode. The CE-ED microchip was demonstrated for the separation of model analytes, including dopamine (DA) and catechol (CA). Detection limits of 132 and 164 nM were achieved for DA and CA, respectively, and a theoretical plate number of 2.5x10(4)/m was obtained for DA. Relative standard deviations in peak heights observed for five runs of a standard solution containing the two analytes (0.1 mM for each) were 1.2 and 3.1% for DA and CA, respectively. The chip could be continuously used for more than 8 h without significant deterioration in analytical performance. PMID- 18393336 TI - Coupling a microchip with electrospray ionization quadrupole time-of-flight mass spectrometer for peptide separation and identification. AB - The present study reports a simple method of coupling a glass microchip to an electrospray ionization (ESI) quadrupole time-of-flight mass spectrometer (QTOF MS) for separation and identification of peptides. A sheath-flow electrospray interface was constructed based on attaching a short fused-silica capillary to the microchip. The dead volume at the interface was effectively reduced by wet etching an approximate flat-bottom capillary insertion channel coaxial to the end of separation microchannel and using a wire-controlled epoxy-blocking attachment method. The makeup liquid and neb gas were coaxially pumped through two stainless steel tees to maintain a stable and efficient electrospray. The coupled microchip/ESI-QTOF-MS system was successfully used to carry out electrophoresis separation of peptides and ESI-QTOF-MS identification. PMID- 18393337 TI - Microfluidic chip of fast DNA hybridization using denaturing and motion of nucleic acids. AB - This study presents the effect of fluidic temperatures and velocities on improving DNA hybridization. The efficiency of hybridization could be improved by introducing elevated temperature in the hot region and velocity in the cold region. Compared with the conventional methods, this hybridization microchip was able to increase the hybridization signal 4.6-fold within 30 min. The 1.4-kb single-stranded target DNA was tested. The increasing tendency of the fluorescence intensity was apparent when the temperature was higher than 82 degrees C, and the fluorescence intensity reached an asymptotic value at T>90 degrees C. A mathematical model was proposed to relate the fluorescence intensity of DNA hybridization with the hot-region temperature and the cold-region velocity. Based on these results, the new hybridization chip with the processes of temperature and velocity differences will improve efficiency of DNA detection. The microchip combined with hot-region temperature and cold-region bulk flow velocity effects could provide additional efficiency in DNA hybridization. PMID- 18393338 TI - Sensitive, label-free protein assay using 1-ethyl-3-methylimidazolium tetrafluoroborate-supported microchip electrophoresis with laser-induced fluorescence detection. AB - Based on the dimer-monomer equilibrium movement of the fluorescent dye Pyronin Y (PY), a rapid, simple, highly sensitive, label-free method for protein detection was developed by microchip electrophoresis with LIF detection. PY formed a nonfluorescent dimer induced by the premicellar aggregation of an anionic surfactant, SDS, however, the fluorescence intensity of the system increased dramatically when proteins such as BSA, bovine hemoglobin, cytochrome c, and trypsin were added to the solution due to the transition of dimer to fluorescent monomer. Furthermore, 1-ethyl-3-methylimidazolium tetrafluoroborate (EMImBF4) instead of PBS was applied as running buffers in microchip electrophoresis. Due to the excellent properties of EMImBF4, not only nonspecific protein adsorption was more efficiently suppressed, but also approximately ten-fold higher fluorescence intensity enhancement was obtained than that using PBS. Under the optimal conditions, detection limits for BSA, bovine hemoglobin, cytochrome c, and trypsin were 1.00x10(-6), 2x10(-6), 7x10(-7), and 5x10(-7) mg/mL, respectively. Thus, without covalent modification of the protein, a protein assay method with high sensitivity was achieved on microchips. PMID- 18393339 TI - On-line cell lysis and DNA extraction on a microfluidic biochip fabricated by microelectromechanical system technology. AB - Integrating cell lysis and DNA purification process into a micrototal analytical system (microTAS) is one critical step for the analysis of nucleic acids. On-chip cell lysis based on a chemical method is realized by sufficient blend of blood sample and the lyzing reagent. In this paper two mixing models, T-type mixing model and sandwich-type mixing model, are proposed and simulation of those models is conducted. Result of simulation shows that the sandwich-type mixing model with coiled channel performs best and this model is further used to construct the microfluidic biochip for on-line cell lysis and DNA extraction. The result of simulation is further verified by experiments. It asserts that more than 80% mixing of blood sample and lyzing reagent which guarantees that completed cell lysis can be achieved near the inlet location when the cell/buffer velocity ratio is less than 1:5. After cell lysis, DNA extraction by means of a solid-phase method is implemented by using porous silicon matrix which is integrated in the biochip. During continuous flow process in the microchip, rapid cell lysis and PCR-amplifiable genomic DNA purification can be achieved within 20 min. The potential of this microfluidic biochip is illustrated by pretreating a whole blood sample, which shows the possibility of integration of sample preparation, PCR, and separation on a single device to work as portable point-of-care medical diagnostic system. PMID- 18393340 TI - Numerical analysis of thermal lens effect for sensitive detection on microchips. AB - Thermal lens microscope (TLM) is a sensitive detection method for nonfluorescent molecules and widely applied to detection in a capillary or on a microchip. In this paper, we developed a flexible design tool for TLM systems to meet various applications utilizing a microspace. The TL effect was modeled, including signal processing, and calculated by combining fluidic dynamics and wave optics software. The coincidence of the calculations and experiments was investigated by measuring the effects of optical path length or focus positions of the excitation beams on TL signals which are quite difficult to calculate by a conventional method. Good agreement was shown and the applicability of the TLM design tool was verified. PMID- 18393341 TI - Study of a novel sample injection method (floating electrokinetic supercharging) for high-performance microchip electrophoresis of DNA fragments. AB - Aiming to achieve high-performance analysis of DNA fragments using microchip electrophoresis, we developed a novel sample injection method, which was given the name of floating electrokinetic supercharging (FEKS). In the method, electrokinetic injection (EKI) and ITP preconcentration of samples was performed in a separation channel, connecting two reservoir ports (P3 and P4) on a cross geometry microchip. At these two stages, side channels, crossing the separation channel, and their ports (P1 and P2) were electrically floated. After the ITP stacked zones passed the cross-part, they were eluted for detection by using leading ions from P1 and P2 that enabled electrophoresis mode changing rapidly from ITP to zone electrophoresis (ZE). Possible sample leakage at the cross-part toward P1 and P2 was studied in detail on the basis of computer simulation using a CFD-ACE+ software and real experiments, through which it was validated that the analyte recovery to the separation channel was almost complete. The FEKS method successfully contributed to higher resolution and shorter analysis time of DNA fragments on the cross-microchip owing to more rapid switching from ITP status to ZE separation in comparison with our previous EKS procedure realized on a single channel microchip. Without any degradation of resolution, the achieved LODs were on average ten times better than using conventional pinched injection. PMID- 18393342 TI - Microchip capillary electrophoresis for frontal analysis of free bilirubin and study of its interaction with human serum albumin. AB - To meet the need for bedside monitoring of free bilirubin for neonates under critical conditions, a microfluidic chip was fabricated and tested for its coupling with CE/frontal analysis (FA) to determine free bilirubin and study of its binding interaction with HSA, which regulated its concentration in plasma. The poly(methyl methacrylate) (PMMA) multichannel chip was fabricated by CO2 laser ablation and bonded with a fused-silica separation capillary for CE/FA separation with UV detection. The chip was designed to allow a complete assay of four electrophoretic runs using preconditioned channels to speed up the determination of free bilirubin and to deliver quick results for bedside monitoring. Under optimized conditions, the linear working range for free bilirubin was from 10 to 200 micromol with RSDs from 2.1 to 5.0% for n=3, and the LOD at 9 micromol for S/N=3. From a binding study between bilirubin and HSA under FA condition, the second binding constant for bilirubin-HSA was determined as 1.07x10(5) L/mol and the number of binding sites per HSA as 3.46. The results enabled the calculation of free bilirubin for jaundiced infants based on the clinically significant level of total bilirubin, producing a range of 118.3-119.4 micromol/L. The developed method is shown to meet the clinical requirement with additional margin of protection to detect the early rising level of free bilirubin prior to jaundice condition. The low-cost microchip CE/FA device is shown to produce quick results with high potential to deliver a suitable bed-side monitoring method for bilirubin management in neonates. PMID- 18393343 TI - A practical interface for microfluidics and nanoelectrospray mass spectrometry. AB - We report a new method for fabricating nanospray ionization tips for MS, formed from glass substrates and the inert polymer, parylene-C. Using a single photolithography step, the emitters are formed contiguously with microchannels, such that no dead volumes are observed. In addition, because the devices are very thin (approximately 0.3 mm) and the tips are formed at rectangular corners, the Taylor cone volumes are small, which makes the method attractive for future integration with microfluidic separations. Device performance was demonstrated by evaluating diverse analytes, ranging from synthetic polymers, to peptides, to nucleic acids. For all analytes, performance was similar to that of conventional emitters (pulled-glass capillaries and the Agilent HPLC Chip) with the advantage of rapid, batch fabrication of identical devices. PMID- 18393344 TI - Accurate quantitation of salivary and pancreatic amylase activities in human plasma by microchip electrophoretic separation of the substrates and hydrolysates coupled with immunoinhibition. AB - A high-performance determination system for alpha-amylase isoenzyme activities in human plasma involving microchip electrophoresis with a plastic chip was developed. The combination of microchip electrophoresis for substrate and hydrolysate separation and an immunoinhibition method for the differentiation of isoenzyme activities using antihuman salivary amylase (S-AMY) mAb allowed the highly selective determination of amylase isoenzyme (S-AMY and pancreatic amylase (P-AMY)) activities even in a complex matrix such as a crude plasma sample. We used 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled maltohexaose (G6) as a substrate. Amylase in a human plasma sample hydrolyzed APTS-G6 into APTS maltotriose (G3) and G3, which was measured as the fluorescence intensity of APTS G3 on microchip electrophoresis. A double logarithm plot revealed a linear relationship between amylase activity and fluorescence intensity in the range of 5-500 U/L of amylase activity (r2=0.9995, p<0.01), and the LOD was 4.38 U/L. Amylase activities in 13 subjects determined by the present method were compared with the results obtained by conventional methods with nitrophenylated oligosaccharides as substrates, respectively. Good correlations were observed for each method on simple linear regression analysis (both p<0.01). The reproducibilities of within-days for total amylase and P-AMY were 2.98-6.27 and 3.83-6.39%, respectively, and these between-days were 2.88-5.66 and 3.64-5.63%, respectively. This system enables us to determine amylase isoenzyme activities in human plasma with high sensitivity and accuracy, and thus will be applicable to clinical diagnosis. PMID- 18393345 TI - Alternating current electrokinetic separation and detection of DNA nanoparticles in high-conductance solutions. AB - In biomedical research and diagnostics, it is a significant challenge to directly isolate and identify rare cells and potential biomarkers in blood, plasma and other clinical samples. Additionally, the advent of bionanotechnology is leading to numerous drug delivery approaches that involve encapsulation of drugs and imaging agents within nanoparticles, which now will also have to be identified and separated from blood and plasma. Alternating current (AC) electrokinetic techniques such as dielectrophoresis (DEP) offer a particularly attractive mechanism for the separation of cells and nanoparticles. Unfortunately, present DEP techniques require the dilution of blood/plasma, thus making the technology less suitable for clinical sample preparation. Using array devices with microelectrodes over-coated with porous hydrogel layers, AC electric field conditions have been found which allow the separation of DNA nanoparticles to be achieved under high-conductance (ionic strength) conditions. At AC frequencies in the 3000 Hz to 10,000 Hz range and 10 volts peak-to-peak, the separation of 10 microm polystyrene particles into low field regions, and 60-nm DNA-derivatized nanoparticles and 200-nm nanoparticles into high-field regions was carried out in 149 mM 1xPBS buffer (1.68 S/m). These results may allow AC electrokinetic systems to be developed that can be used with clinically relevant samples under physiological conditions. PMID- 18393346 TI - Luminescent alkynylplatinum(II) complexes of 2,6-bis(N-alkylbenzimidazol-2' yl)pyridine-type ligands with ready tunability of the nature of the emissive states by solvent and electronic property modulation. AB - A new class of luminescent alkynylplatinum(II) complexes of tridentate bis(N alkylbenzimidazol-2'-yl)pyridines (bzimpy), [Pt(R,R'-bzimpy)(C[triple chemical bond]C-R'')]X (X=PF(6), OTf), and one of their chloro precursor complexes, [Pt(R,R'-bzimpy)Cl]PF(6), have been synthesized and characterized; one of the alkynyl complexes has also been structurally characterized by X-ray crystallography. Electrochemical studies showed that the oxidation wave is alkynyl ligand-based in nature with some mixing of the metal center-based contribution, whereas the two quasi-reversible reduction couples are mainly bzimpy-based reductions. The electronic absorption and luminescence properties of the complexes have also been investigated. In solution, the high-energy and intense absorption bands are assigned as the pi-pi* intraligand (IL) transitions of the bzimpy and alkynyl ligands, whereas the low-energy and moderately intense absorptions are assigned to an admixture of metal-to-ligand charge-transfer (MLCT) (dpi(Pt)-->pi*(R,R'-bzimpy)) and ligand-to-ligand charge-transfer (LLCT) (pi(C[triple chemical bond]C-R'')-->pi*(R,R'-bzimpy)) transitions. Upon variation of the electronic effects of the arylalkynyl ligands, vibronic-structured or structureless emission bands, originating from triplet metal-perturbed intraligand (IL) or an admixture of triplet metal-to-ligand charge-transfer (MLCT) and ligand-to-ligand charge-transfer (LLCT) excited states respectively, were observed in solution. Interestingly, two of the complexes showed a dual luminescence that was sensitive to the polarity of the solvents. Upon cooling from 298 K to 155 K, drastic color, UV/Vis, and luminescence changes were observed in a butyronitrile solution of 1, and were ascribed to the formation of aggregate species through PtPt and pi-pi stacking interactions. DFT and time dependent DFT (TD-DFT) calculations have been performed to verify and elucidate the results of the electrochemical and photophysical properties. PMID- 18393347 TI - Folding of coordination polymers into double-stranded helical organization. AB - Self-assembling coordination polymers based on Pd II and Cu II metal ions were prepared from complexation of a bent-shaped bispyridine ligand and a corresponding transition metal. These coordination polymers were observed to self assemble into supramolecular structures that differ significantly depending on the coordination geometry of the metal center. The polymer based on Pd II self assembles into a layer structure formed by bridging bispyridine ligands connected in a trans-position of the square-planar coordination geometry of metal center. In contrast, the polymer based on Cu II adopts a double-helical conformation with regular grooves, driven by interstranded, copper-chloride dimeric interaction. The double-stranded helical organization is further confirmed by structure optimization from density functional theory with aromatic framework, showing that the optimized double-helical structure is energetically favorable and consistent with the experimental results. These results demonstrate that weak metal-ligand bridging interactions can provide a useful strategy to construct stable double stranded helical nanotubes. PMID- 18393348 TI - Polyelectrolyte-induced self-assembly of positively charged alkynylplatinum(II) terpyridyl complexes in aqueous media. AB - Polyelectrolytes carrying multiple negative charges were found to induce the aggregation and self-assembly of the positively charged platinum(II)-terpyridyl complexes in aqueous media. The aggregation and self-assembly of the complexes were driven by electrostatic interactions between the polymer and the complex, and by terpyridine ligand pi-pi stacking and platinum-platinum (metal-metal) interactions. As a result, remarkable UV/Vis and emission spectral changes were observed. The spectroscopic property changes were related to the structural properties of the metal complexes as well as the polyelectrolytes. The induced self-assembly of the platinum complexes was also strongly affected by the solution properties of the aqueous media, for example, the solution pH, ionic strength, and the percentage of organic solvent added. PMID- 18393349 TI - Synthesis of a paramagnetic polymer by ring-opening polymerization of a strained [1]vanadoarenophane. PMID- 18393350 TI - A model testing factors that influence physical activity for Taiwanese adults with anxiety. AB - The aim of this study was to test a theoretical model of the facilitators and barriers to physical activity for Taiwanese adults with anxiety. Structural equation modeling was used to examine both one-way direct and indirect influences of 11 personal and cognitive-emotional factors on physical activity in this population (N = 239). The final version of the model provided a good fit to the data, with nine variables explaining 23.3% of the variance in physical activity. Perceived life stress events, perceived benefits of activity, and perceived self efficacy for activity directly influenced physical activity. The results suggest that these variables should be addressed in designing treatment physical activity programs for Taiwanese individuals with anxiety. PMID- 18393351 TI - Bone morphogenetic protein-7 regulates differentially the mRNA expression of bone morphogenetic proteins and their receptors in rat achilles and patellar tendon cell cultures. AB - Previous animal studies have suggested that certain bone morphogenetic proteins (BMPs) may be useful therapeutically in treating tendon healing. To better understand the relationship among the different BMPs in the healing process, we initiated the present study to examine the effects of a member of the BMP family, BMP-7 (also called Osteogenic Protein-1) on the temporal and spatial expression patterns of other BMPs and the BMP receptors in cell cultures of adult rat Achilles and Patellar tendons. Cultures from both tendon types expressed detectable but variable levels of biochemical markers characteristics of tendons. RNAs coding for type II collagen and transcription factors Six1, Scleraxis, and Tendin were detected in both types of cultures. Distinct patterns of expression of several BMP members and their receptors were observed in these cultured cells and BMP-7 exerted differential effects on their expression. The findings may have implications in the treatment of different tendon injuries with BMPs. PMID- 18393352 TI - Paternal contributions: new functional insights for spermatozoal RNA. AB - Whereas the presence of RNA in mature ejaculate spermatozoa is now established, its functional significance, if any, is still a matter of debate. This reflects the accepted description that spermatozoa are highly differentiated, specialized cells of minimal cytoplasm and compacted nucleus that are transcriptionally inactive. A significant proportion of the RNA required for the later, haploid stages of terminal spermatogenic differentiation (spermiogenesis) is synthesized prior to transcriptional arrest then stably stored until its translation during spermiogenesis. Spermatozoal RNAs, including messenger RNAs (mRNAs) are therefore considered to be stored remnants. Any role in fertilization and early development has, until recently, seemed unlikely, since the oocyte contains large stores of maternal mRNAs known to be required for early embryonic development prior to zygotic genome activation. Although the spermatozoon can deliver its RNA to the oocyte at fertilization, it has been generally assumed that compared to the oocyte RNA reserve, the spermatozoan payload is too small to be functional in embryo development. However, the debate continues as recent studies suggest that in specific instances sperm RNA is functional. This review presents and discusses the functional significance of spermatozoal RNA in relation to some recent advances in the field. PMID- 18393353 TI - A novel butyrolactone derivative inhibited smooth muscle cell migration and proliferation and maintained endothelial cell functions through selectively affecting Na, K-ATPase activity and mitochondria membrane potential during in vitro angiogenesis. AB - We have found that 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran -2(3H)-one (3BDO), could effectively suppress human umbilical vascular endothelial cell (HUVEC) apoptosis induced by deprivation of fibroblast growth factor-2 and serum. Here, our purpose was to investigate whether 3BDO could modulate angiogenesis and its possible acting mechanism. The effect of 3BDO on angiogenesis was investigated by capillary-like tubule formation and rat aortic ring assay. Proliferation and migration of cells were detected by counting living cell number and scraping cell monolayer, respectively. Na, K-ATPase activity was measured spectrophotometrically. Mitochondrial membrane potential was analyzed using tetramethylrhodamine methylester fluorescence by confocal microscopy. Our results showed that 3BDO inhibited migration and proliferation of vascular smooth muscle cells (VSMCs), but maintained migration and tubule formation of HUVECs. In HUVECs, 3BDO inhibited Na, K-ATPase activity, but had no effect on mitochondria membrane potential. In VSMCs, it did not affect Na, K-ATPase activity, but depressed mitochondria membrane potential obviously. The data showed that 3BDO had selective effects on HUVECs and VSMCs, it might perform its role through the selective effects on the activity of Na, K-ATPase and the mitochondria membrane potential in HUVECs and VSMCs. PMID- 18393354 TI - Extended high-frequency audiometry and noise induced hearing loss in cement workers. AB - BACKGROUND: It has been suggested that extended high-frequency audiometry (EHFA) might be more sensitive than conventional audiometry in detecting early signs of hearing impairment. However, this technique has not been adequately tested in an occupational environment. We therefore investigated the usefulness of this method in noise-exposed workers. METHODS: We compared conventional frequency audiometry (0.25-8 kHz) and EHFA (9-18 kHz) in 184 noise-exposed and 98 non-noise-exposed workers. RESULTS: Both methods showed significantly higher threshold levels (P < 0.05) in noise-exposed workers for most of the tested frequencies; however, the differences were more marked for EHFA, especially in young exposed workers. Significant differences in the EHF range were detected also in the subgroup of noise-exposed workers with normal findings at conventional audiometry. Stepwise regression analysis showed that in 21- to 40-year-old workers the noise effect was largely predominant at both conventional audiometry and EHFA, whereas in older subjects the noise effect was predominant up to 6 kHz frequency, the effect of age being significantly greater at higher frequencies. CONCLUSIONS: These data indicate that EHFA is more sensitive than conventional audiometry in detecting noise induced hearing loss. However, hearing loss in the EHF range seems an age dependent phenomenon with progression into the lower speech range frequencies with increasing age. These changes seem to be accentuated in the early years by noise exposure, suggesting that EHFA could represent a useful preventive measure in young exposed workers. PMID- 18393355 TI - Coactivators and nuclear receptor transactivation. AB - A variety of coregulator proteins serve as partners for nuclear receptors orchestrating the molecular events required for receptor-dependent transcriptional regulation. Some coregulators directly interact with nuclear receptors and provide a platform for recruitment of other factors that provide distinct biochemical activities that influence transcriptional efficiency. Coregulators can influence chromatin structure and activity via direct modification of histone proteins or by facilitating ATP-dependent chromatin remodeling. They also have the capacity to impact multiple steps in the transcription process including initiation, elongation, and mRNA splicing. Genetic analysis in humans and animal models are revealing the important cell and tissue-type specific actions of nuclear receptor coregulators as well and their role in human physiology and disease. PMID- 18393356 TI - Small applied electric fields guide migration of hippocampal neurons. AB - Effectively directed neuron migration is critical for development and repair in the central nervous system (CNS). Endogenous electric fields (EFs) are widespread in developing and regenerating tissues and regulate a variety of cell behaviors including directed cell migration. Electrically-directed neuronal migration has not been tested previously and we show that an applied EF directs migration of hippocampal neurons toward the cathode at a field strength of 120 mV/mm, close to the physiological range. Reversal of the field polarity reversed the direction of neuron migration. Neuron migration from an explant also was directed by an applied EF. Mechanistically, EF-guided migration was transduced by activation of the second messenger molecules ROCK (Rho-associated protein kinase) and PI3 kinase (phosphoinositide-3 kinase) since their pharmacological inhibition decreased the directedness and speed of neuron migration. This work demonstrates that rat hippocampal neurons respond to applied EFs with directional migration and raises the possibility that EFs may be used as a cue to direct neuronal migration in novel strategies to repair the CNS. PMID- 18393357 TI - Chimpanzee (Pan troglodytes) birth patterns and human presence in zoological settings. AB - In response to work demonstrating a negative correlation between human staff activity and parturition in laboratory-managed primates, this study examined the distribution of 231 captive chimpanzee (Pan troglodytes) births that occurred in accredited American zoological institutions by the day of the week on which the birth was discovered. We hypothesized that if chimpanzee parturition patterns displayed sensitivity to human presence, then fewer births per day would be reported during the weekend period, when the visitor density was high, as compared with the lower density working week. Analyses indicated that chimpanzee births were randomly distributed throughout the week. In the context of the questionable sensitivity of primate parturition to external influence, results suggest that variations in human presence do not affect the fine-level timing of birth in chimpanzees managed in a zoological setting. PMID- 18393358 TI - Characterization and functional studies of a FYVE domain-containing phosphatase in C. elegans. AB - The myotubularin (MTM) enzymes are phosphatidylinositol 3-phosphate (PI3P) and phosphatidylinositol 3,5-bisphosphate phosphatases. Mutation of MTM1, the founder member of this family, is responsible for X-linked myotubular myopathy in humans. Here, we have isolated and characterized a Caenorhabditis elegans homology of the enzymes designated ceMTM3. ceMTM3 preferably dephosphorylates PI3P and contains a FYVE lipid-binding domain at its C-terminus which binds PI3P. Immunoblotting analyses revealed that the enzyme is expressed during the early development and adulthood of the animal. Immunofluorescent staining revealed predominant expression of the enzyme in eggs and muscles. Knockdown of the enzyme by using feeding-based RNA interference resulted in an increased level of PI3P and caused severe impairment of body movement of the worms at their post-reproductive ages and significantly shortened their lifespan. This study thus reveals an important role of the MTM phosphatases in maintaining muscle function, which may have clinical implications in prevention and treatment of sarcopenia. PMID- 18393359 TI - Apoptosis in arsenic trioxide-treated Calu-6 lung cells is correlated with the depletion of GSH levels rather than the changes of ROS levels. AB - Arsenic trioxide (ATO) can regulate many biological functions such as apoptosis and differentiation in various cells. We investigated an involvement of ROS such as H(2)O(2) and O(2)(*-), and GSH in ATO-treated Calu-6 cell death. The levels of intracellular H(2)O(2) were decreased in ATO-treated Calu-6 cells at 72 h. However, the levels of O(2)(*-) were significantly increased. ATO reduced the intracellular GSH content. Many of the cells having depleted GSH contents were dead, as evidenced by the propidium iodine staining. The activity of CuZn-SOD was strongly down-regulated by ATO at 72 h while the activity of Mn-SOD was weakly up regulated. The activity of catalase was decreased by ATO. ROS scavengers, Tiron and Trimetazidine did not reduce levels of apoptosis and intracellular O(2)(*-) in ATO-treated Calu-6 cells. Tempol showing a decrease in intracellular O(2)(*-) levels reduced the loss of mitochondrial transmembrane potential (DeltaPsi(m)). Treatment with NAC showing the recovery of GSH depletion and the decreased effect on O(2)(*-) levels in ATO-treated cells significantly inhibited apoptosis. In addition, BSO significantly increased the depletion of GSH content and apoptosis in ATO-treated cells. Treatment with SOD and catalase significantly reduced the levels of O(2)(*-) levels in ATO-treated cells, but did not inhibit apoptosis along with non-effect on the recovery of GSH depletion. Taken together, our results suggest that ATO induces apoptosis in Calu-6 cells via the depletion of the intracellular GSH contents rather than the changes of ROS levels. PMID- 18393360 TI - c-Myc represses FOXO3a-mediated transcription of the gene encoding the p27(Kip1) cyclin dependent kinase inhibitor. AB - The p27(Kip1) (p27) cyclin-dependent kinase inhibitor and c-Myc oncoprotein play essential roles in control of cell cycle progression and apoptosis. Induction of p27 (CDKN1B) gene transcription by Forkhead box O proteins such as FOXO3a leads to growth arrest and apoptosis. Previously, we observed that B cell receptor (surface IgM) engagement of WEHI 231 immature B lymphoma cells with an anti-IgM antibody results in activation of FOXO3a, growth arrest and apoptosis. As ectopic c-Myc expression in these cells prevented anti-IgM induction of p27 and cell death, we hypothesized that c-Myc represses FOXO3a-mediated transcription. Here we show that c-Myc inhibits FOXO3a-mediated activation of the p27 promoter in multiple cell lines. The mechanism of this repression was explored using a combination of co-immunoprecipitation, oligonucleotide precipitation, and chromatin immunoprecipitation experiments. The studies demonstrate a functional association of FOXO3a and c-Myc on a proximal Forkhead binding element in the p27 promoter. This association involves the Myc box II domain of c-Myc and the N terminal DNA-binding portion of FOXO3a. Analysis of publicly available microarray datasets showed an inverse pattern of c-MYC and p27 RNA expression in primary acute myeloid leukemia, prostate cancer and tongue squamous cell carcinoma samples. The inhibition of FOXO3a-mediated activation of the p27 gene by the high aberrant expression of c-Myc in many tumor cells likely contributes to their uncontrolled proliferation and invasive phenotype. PMID- 18393361 TI - Molecular parameters of genome instability: roles of fragile genes at common fragile sites. AB - Common chromosome fragile sites occur at specific sequences within mammalian genomes that exhibit apparent single-stranded regions in mitotic chromosomes on exposure of cells to replication stress. Recent progress in the characterization of sequences, and more precise mapping of common fragile sites in mammalian and yeast genomes, has led to the exact placement of large common fragile regions straddling the borders of chromosomal G and R bands, with early and late replicating genomic regions, respectively, and could lead to breakthroughs in understanding the function of these evolutionarily conserved but highly recombinogenic chromosome elements. Deficiency of genes involved in DNA damage checkpoint responses, such as ATR, CHK1, HUS1 leads to increased frequency of fragile site instability. Some of these fragile sites, particularly FRA3B, encode genes that are themselves involved in the protection of cells from DNA damage through various mechanisms. Protection of mammalian genomes from accumulation of DNA damage in somatic cells is critical during development, puberty and during the reproductive lifespan, and occurs through mechanisms involving surveillance of the genome for damage, signals to the cell cycle machinery to stop cell cycle progression, signals to repair machinery to repair damage, signals to resume cycling or initiate apoptotic programs, depending on the extent of damage and repair. When genes involved in these processes are altered or deleted, cancer can occur. The tumor suppressor gene, FHIT at the FRA3B locus, and possibly other fragile genes, is a common target of damage and paradoxically encodes a protein with roles in protection from DNA damage. PMID- 18393362 TI - Autophagy in the liver. AB - A great part of our current understanding of mammalian macroautophagy is derived from studies of the liver. The term "autophagy" was introduced by Christian de Duve in part based on ultrastructural changes in rat liver following glucagon injection. Subsequent morphological, biochemical, and kinetics studies of autophagy in the liver defined the basic process of autophagosome formation, maturation, and degradation and the regulation of autophagy by hormones, phosphoinositide 3-kinases, and mammalian target of rapamycin. It is now clear that macroautophagy in the liver is important for the balance of energy and nutrients for basic cell functions, the removal of misfolded proteins resulting from genetic mutations or pathophysiological stimulations, and the turnover of major subcellular organelles such as mitochondria, endoplasmic reticulum, and peroxisomes under both normal and pathophysiological conditions. Disturbance of autophagy function in the liver could thus have a major impact on liver physiology and liver disease. PMID- 18393364 TI - Sustained complete response and low complication rates after radiofrequency ablation of very early hepatocellular carcinoma in cirrhosis. PMID- 18393365 TI - Identification of the Alzheimer's disease amyloid precursor protein (APP) and its homologue APLP2 as essential modulators of glucose and insulin homeostasis and growth. AB - The amyloid precursor protein (APP), the source of the neurotoxic amyloid beta (A beta) peptide involved in Alzheimer's disease (AD), belongs to a conserved family of related proteins. In mammals, the APP family contains amyloid precursor-like protein 1 (APLP1) and amyloid precursor-like protein 2 (APLP2). Whilst a number of activities have been attributed to the APP family, an overall function has not been definitively established. While ablating either the APP or APLP2 gene in mice produces minimal phenotypic change, the combined knockout of these genes in mice causes postnatal mortality. Postnatal survival therefore requires a shared but unknown function of APP and APLP2. To investigate the biochemical basis for the postnatal lethality, plasma was analysed from double knockout mice (APP-/- APLP2-/-) 2 days before birth, at gestational day E17, and from mice at 12-16 h after birth. The postnatal double knockouts had 66% lower plasma glucose levels than their wild-type controls and 50% lower than their single knockout counterparts. Interestingly, the postnatal double knockouts displayed hyperinsulinaemia, as shown by inappropriate plasma insulin levels, given their degree of hypoglycaemia. The single knockout mice also showed hyperinsulinaemia and had 31% lower plasma glucose than the wild-types. While the double knockouts did not survive more than 24 h after birth, the single knockouts reached adulthood and their hypoglycaemia continued. Therefore, APP and APLP2 expression modulates plasma insulin and glucose concentrations. Plasma calcium, magnesium and phosphate were also significantly reduced in the double knockouts compared to the wild-types, and they showed distinctive growth restriction, suggesting the involvement of a metabolic impairment. These results link the expression of the APP and APLP2 genes with glucose homeostasis and growth and therefore identify a novel function for the APP family. PMID- 18393366 TI - Most multifocal papillary thyroid carcinomas acquire genetic and morphotype diversity through subclonal evolution following the intra-glandular spread of the initial neoplastic clone. AB - Papillary thyroid carcinoma (PTC) is frequently multifocal (mPTC), with synchronous tumour foci often showing varied morphology. The genetic mechanisms underlying the development of multiple and histologically diverse tumour foci remain uncertain. Different tumour foci might develop either through intrathyroidal dissemination of a single malignant clone, with morphotype differentiation occurring as a result of subclonal progression, or they may stem from independent transformational events involving multiple progenitor clones. To determine the clonal derivation of multiple tumour foci and to map their clonal relationships and genetic progression in mPTC, we evaluated genome-wide allelic imbalances (AI) and BRAF V600E mutation status in 55 synchronous tumour foci from 18 mPTC patients. For apparently monoclonal tumours, we calculated the probabilities of monoclonal derivation and used phylogenetic analysis to model clonal evolution. Genome-wide allelotyping and BRAF mutation analysis showed genetic alterations consistent with monoclonal origin in 83% of cases, mostly with evidence of subclonal evolution. BRAF V600E mutations were early events during clonal evolution of most, but not all cases. MPTC with morphologically diverse tumour foci also arose through monoclonal derivation in 75% of cases, demonstrating that morphotype-determining genetic changes can be acquired during clonal diversification, subsequent to the spread of the original malignant progenitor clone. In 17% of patients, discordant AI or BRAF V600E profiles implied that mPTCs can occasionally develop from distinct transformation events. This study suggests that mPTC originates usually from neoplastic transformation and subsequent intrathyroidal spread of a single malignant progenitor clone. Clonal progression and morphotype differentiation occur through progressive acquisition of genetic alterations subsequent to the initial intra-glandular spread. In monoclonal BRAF V600E-positive mPTCs, BRAF V600E is not always present in all tumour foci, indicating that other tumour-genetic factors in the primary progenitor clone can also trigger PTC neoplastic transformation. PMID- 18393367 TI - Cadherin switching dictates the biology of transitional cell carcinoma of the bladder: ex vivo and in vitro studies. AB - Bladder cancer is the fifth most common malignancy in the UK. Clinically, the most important process in determining prognosis is the development of invasion, initially of the lamina propria and then beyond as these transitional cell carcinomas (TCCs) progress from stage pT1 to stages T2+. Cadherins and catenins are the main mediators of cell-cell interactions in epithelial tissues, and loss of membranous E-cadherin immunoreactivity is strongly correlated with high grade, advanced stage and poor prognosis in bladder cancer and other malignancies. However, the role of P-cadherin is yet to be fully elucidated in bladder TCC. The objectives of this study were to establish how the expression of cadherins and catenins determines clinical and in vitro behaviour in bladder TCC. Utilizing immunohistochemistry, immunofluorescence and western blotting, we demonstrated a significant reduction in the expression of E-cadherin and beta-catenin as grade and stage of bladder TCC progress, accompanied by a significant increase in P cadherin expression (all p < 0.05, Pearson's chi2 test). Increased P-cadherin expression was also associated with a significantly worse bladder cancer-specific survival (log rank p = 0.008), with Cox regression showing P-cadherin to be an independent prognostic factor. Utilizing a variety of tissue culture models in a range of functional studies, we demonstrated that P-cadherin mediates defective cell-cell adhesion and enhances anchorage-independent growth. The results provide evidence that increased P-cadherin expression promotes a more malignant and invasive phenotype of bladder cancer, and appears to have a novel role late in the disease. PMID- 18393368 TI - Over-expression of polycomb group protein EZH2 relates to decreased expression of p16 INK4a in cholangiocarcinogenesis in hepatolithiasis. AB - Polycomb group protein EZH2 and Bmi1 are reportedly involved in the progression of malignant tumours. We examined the participation of EZH2 in multi-step cholangiocarcinogenesis in hepatolithiasis with respect to tumour suppressor gene p16 INK4a. We examined 20 hepatolithiatic livers with intrahepatic cholangiocarcinoma (ICC) and 10 histologically normal livers. Neoplastic biliary lesions were classified into biliary intraepithelial neoplasm (BilIN-1, 2 and 3) and invasive carcinoma. We selected 15 foci of invasive carcinoma, 8 BilIN-3 (carcinoma in situ), 12 BilIN-2 (high-grade dysplasia), 32 BilIN-1 (low-grade dysplasia) and 37 non-neoplastic biliary epithelia from these livers. Expression of p16 INK4a, EZH2 and Bmi1 were surveyed in these foci. P16 INK4a promoter methylation was examined in microdissected tissues. Taking advantage of two cell lines of CC (HuCTT-1 and TFK-1) and small interfering RNA (siRNA), the effects of the knockdown of EZH2 on p16 INK4a methylation of CC cells were examined. Expression of p16 INK4a, which was frequent in BilIN1, was decreased in BilIN-2/3 and invasive carcinoma, while EZH2 expression showed step-wise increase from BilIN-1, -2 and -3 to invasive carcinoma (p < 0.01). P16 INK4a promoter hypermethylation was related to aberrant expression of EZH2. The knockdown of EZH2 in cultured CC cells decreased p16 INK4a methylation and decreased binding of EZH2 to the p16 INK4a gene promoter. The latter suggested that direct binding of EZH2 is involved in the regulation of the p16 INK4a gene. Our data suggest that over-expression of EZH2 may induce hypermethylation of p16 INK4a promoter followed by decreased expression of p16 INK4a in the multi-step cholangiocarcinogenesis through intraepithelial neoplasm in hepatolithiasis. PMID- 18393369 TI - Monitoring of epithelial cell caspase activation via detection of durable keratin fragment formation. AB - Keratins 18 and 19 (K18/K19) are epithelial-specific intermediate filament proteins. Apoptosis induces caspase cleavage at the highly conserved K18 or K19 Asp237, which in K18 is preceded by cleavage at Asp396. We characterized the keratin N-terminal fragments that are generated upon caspase digestion of K18/K19 at Asp237 in order to study keratin dynamics during apoptosis. This was carried out by generating and characterizing antibodies selective to K18/K19 Asp237. K18 or K19 peptides that expose Asp237 in 234VEVD were used for rabbit immunization. The generated antibodies recognized cleaved but not intact K18/K19, exclusively, as determined by blotting or immunofluorescence staining of apoptotic human HT29 cells or livers isolated from Fas-Ab-injected mice. Antibodies to K18/K19 Asp237 recognized the common VEVD-motif as determined by immunoblotting of cells transfected with K18, K19 or K20. The K18/K19 VEVD-directed antibodies demonstrated sequential Asp396 then Asp237 K18 cleavage during apoptosis. Specific-keratin selectivity of the anti-Asp237 antibodies was confirmed by their inability to recognize K14 after UV-induced apoptosis in transfected cells. The Asp237-containing apoptotic keratin fragments are secreted into the medium of cultured HT29 cells and are stable up to 96 h after inducing apoptosis. Furthermore, the generated antibodies recognize keratin apoptotic fragments in sera of mice undergoing hepatocyte apoptosis and sera of patients with cirrhosis, and also recognize apoptotic cells in various epithelial human tumours. Therefore, the N-terminal caspase-generated K18 fragment is stable in tissues and biological fluids. The Asp237-directed antibodies provide a powerful tool to study apoptosis in human and mouse tissues, cells and serum, using a broad range of detection modalities. PMID- 18393371 TI - The role of Hfe in transferrin-bound iron uptake by hepatocytes. AB - HFE-related hereditary hemochromatosis results in hepatic iron overload. Hepatocytes acquire transferrin-bound iron via transferrin receptor (Tfr) 1 and Tfr1-independent pathways (possibly Tfr2-mediated). In this study, the role of Hfe in the regulation of hepatic transferrin-bound iron uptake by these pathways was investigated using Hfe knockout mice. Iron and transferrin uptake by hepatocytes from Hfe knockout, non-iron-loaded and iron-loaded wild-type mice were measured after incubation with 50 nM (125)I-Tf-(59)Fe (Tfr1 pathway) and 5 microM (125)I-Tf-(59)Fe (Tfr1-independent or putative Tfr2 pathway). Tfr1 and Tfr2 messenger RNA (mRNA) and protein expression were measured by real-time polymerase chain reaction and western blotting, respectively. Tfr1-mediated iron and transferrin uptake by Hfe knockout hepatocytes were increased by 40% to 70% compared with iron-loaded wild-type hepatocytes with similar iron levels and Tfr1 expression. Iron and transferrin uptake by the Tfr1-independent pathway was approximately 100-fold greater than by the Tfr1 pathway and was not affected by the absence of Hfe. Diferric transferrin increased hepatocyte Tfr2 protein expression, resulting in a small increase in transferrin but not iron uptake by the Tfr1-independent pathway. CONCLUSION: Tfr1-mediated iron uptake is regulated by Hfe in hepatocytes. The Tfr1-independent pathway exhibited a much greater capacity for iron uptake than the Tfr1 pathway but it was not regulated by Hfe. Diferric transferrin up-regulated hepatocyte Tfr2 protein expression but not iron uptake, suggesting that Tfr2 may have a limited role in the Tfr1-independent pathway. PMID- 18393372 TI - S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation. AB - We previously showed that S-adenosylmethionine (SAMe) and its metabolite methylthioadenosine (MTA) blocked lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNFalpha) expression in RAW (murine macrophage cell line) and Kupffer cells at the transcriptional level without affecting nuclear factor kappa B nuclear binding. However, the exact molecular mechanism or mechanisms of the inhibitory effect were unclear. While SAMe is a methyl donor, MTA is an inhibitor of methylation. SAMe can convert to MTA spontaneously, so the effect of exogenous SAMe may be mediated by MTA. The aim of our current work is to examine whether the mechanism of SAMe and MTA's inhibitory effect on proinflammatory mediators might involve modulation of histone methylation. In RAW cells, we found that LPS induced TNFalpha expression by both transcriptional and posttranscriptional mechanisms. SAMe and MTA treatment inhibited the LPS-induced increase in gene transcription. Using the chromatin immunoprecipitation assay, we found that LPS increased the binding of trimethylated histone 3 lysine 4 (H3K4) to the TNFalpha promoter, and this was completely blocked by either SAMe or MTA pretreatment. Similar effects were observed with LPS-mediated induction of inducible nitric oxide synthase (iNOS). LPS increased the binding of histone methyltransferases Set1 and myeloid/lymphoid leukemia to these promoters, which was unaffected by SAMe or MTA. The effects of MTA in RAW cells were confirmed in vivo in LPS treated mice. Exogenous SAMe is unstable and converts spontaneously to MTA, which is stable and cell-permeant. Treatment with SAMe doubled intracellular MTA and S adenosylhomocysteine (SAH) levels. SAH also inhibited H3K4 binding to TNFalpha and iNOS promoters. CONCLUSION: The mechanism of SAMe's pharmacologic inhibitory effect on proinflammatory mediators is mainly mediated by MTA and SAH at the level of histone methylation. PMID- 18393373 TI - Which patients with IBD need psychological interventions? A controlled study. AB - BACKGROUND: Psychological distress is frequent in inflammatory bowel disease (IBD). Whether there is a need for psychological interventions is unknown. This study investigated the quantity and quality of the need for psychological interventions in IBD as compared to rheumatoid arthritis (RA). METHODS: In all, 302 patients with IBD and 109 patients with RA answered the ADAPT questionnaire, assessing the need for psychosomatic support (physicians support) and for psychotherapy, the hospital anxiety and depression scale, the SF-36, a questionnaire on social support (SOZU-K22), and the Rating Form of IBD Patient Concerns (IBD patients only). Detailed biomedical data were also assessed. RESULTS: Ninety-three patients with IBD (31%) expressed a need for psychological intervention compared to 14 patients with RA (13%; P < 0.001). Stepwise logistic regression analysis revealed that anxiety (odds ratio [OR] 3.6; 95% confidence interval [CI] 2.2-6.0; P < 0.001), age < or =44 years (OR 2.6; 95% CI 1.5-4.3; P < 0.001) and impaired social support (SOZU-K22 <4.20) (OR 2.0; 95% CI 1.2-3.3; P = 0.009) accounted for this difference. In IBD the need for psychosomatic (physicians) support was associated with worries and concerns about IBD and the need for psychotherapy was associated with worries and concerns about IBD, anxiety, impaired "social functioning" (SF-36), and short disease duration. CONCLUSIONS: Patients with IBD express a higher need for psychological interventions than patients with RA due to greater psychosocial restrictions inherent in IBD. The need for psychological interventions was characterized by psychological factors, mainly worries about the disease and anxiety, rather than by medical variables PMID- 18393374 TI - Ulcerative colitis surveillance: activity does matter. PMID- 18393375 TI - Efficacy of infliximab for induction and maintenance of remission in intestinal Behcet's disease. AB - BACKGROUND: Intestinal Behcet disease (BD) is characterized by intestinal inflammation with round and oval ulcers associated with gastrointestinal symptoms. Although several cases have been reported that infliximab is effective for induction of remission, the efficacy of infliximab for maintaining remission is unknown. METHODS: Six cases with fulminant intestinal BD were treated with infliximab. All patients were steroid-dependent and refractory to immunosuppressants; 3 patients were treated with 6-mercaptopurine, 1 patient with azathioprine, 1 patient with cyclosporine A, and 1 patient with methotrexate. RESULTS: Four patients achieved remission by infliximab and all of these patients maintained remission with scheduled treatments of infliximab, with the longest duration of remission being about 3 years. Another 2 patients with ileal ulceration required surgery; however, 1 patient has maintained remission by scheduled treatment of infliximab for 2 years after surgery. CONCLUSIONS: Infliximab appears to offer an option for fulminant intestinal BD to induce and maintain remission, although a randomized control trial is needed. PMID- 18393376 TI - To lap or not to lap, that is the question...no longer? PMID- 18393377 TI - Lymphocyte homing and its role in the pathogenesis of IBD. AB - Inflammatory bowel disease (IBD) is an idiopathic disorder of chronic inflammation of the gastrointestinal tract. Experimental models of IBD and results from human genomewide linkage studies suggest that the primary defect that leads to IBD is an inappropriate mucosal immune response to normal intestinal microbes. Genetic alterations not only confer increased susceptibility to IBD but also appear to determine the nature and location of the intestinal inflammation, as is evident in patients with genetic alterations of NOD2 and their susceptibility for ileal Crohn's disease. IBD has traditionally been classified into 2 subtypes, namely, ulcerative colitis (UC) and Crohn's disease (CD), based on histological appearance and anatomical distribution. However, an increasing body of data supports the concept that IBD is an umbrella diagnosis encompassing a variety of disorders with distinct genetic, microbial, and environmental determinants that cluster either into a UC or CD phenotype. The shared common pathway is uncontrolled intestinal inflammation. A key element in the pathogenesis of intestinal inflammation in both UC and CD is increased leukocyte recruitment from the circulation, and this provides a potential target for pharmaceutical inhibition. In this article we review the current understanding of the molecules that determine leukocyte trafficking to the gut and highlight opportunities where their inhibition could be exploited to treat IBD.(Inflamm Bowel Dis 2008). PMID- 18393378 TI - Contrasting cytoskeletal regulation of MHC class II peptide presentation by human B cells or dendritic cells. AB - MHC class II-mediated antigen presentation by B lymphocytes or dendritic cells (DC) initiates CD4+ T lymphocyte activation. In B lymphocytes, MHC class II peptide presentation has been characterised by recruitment of MHC class II, F actin and lipid rafts to the B cell-T cell immunological synapse. We now show that MHC class II engagement in B lymphocytes induced lipid raft-independent Rho and Rac activation and that inhibition of either Rho-GTPase activation or actin polymerisation in the B cell abrogated T cell activation without altering B cell T cell conjugate formation. Short-hairpin RNA studies excluded a role for the Cdc42 effector Wiskott-Aldrich syndrome protein. In contrast, antigen presentation by DC was Rho-GTPase-independent although actin was recruited to the DC-T cell interaction site. Moreover, actin depolymerisation in the DC significantly increased T cell activation without altering the number of DC-T cell conjugates. Finally we show that stable recruitment of HLA-DR to the site of the immunological synapse is not a uniform observation in DC and demonstrate reduced HLA-DR expression at the site of microtubule organising centre polarization. Therefore although actin accumulates in DC and B lymphocytes at the immunological synapse with antigen-specific T lymphocytes, this does not reflect comparable functional roles of their actin cytoskeletons in antigen presentation. PMID- 18393379 TI - First-trimester diagnosis of cesarean scar ectopic pregnancy. AB - Once considered extremely rare, implantation of a pregnancy within the scar of a previous cesarean section is becoming more common. In fact, its incidence is now higher than that of cervical ectopic pregnancies. We identified 5 cases of ectopic pregnancy implanted in a prior cesarean section scar at our institution since 2004. We outline the criteria for the first-trimester sonographic diagnosis of cesarean scar ectopic pregnancy, including a new sign of lower uterine segment ballooning, which has previously not been reported. Clinicians must have a heightened awareness of this serious and potentially fatal pregnancy complication. PMID- 18393380 TI - Duodenal duplication cyst: a rare cause of acute pancreatitis in children. AB - Duodenal duplication is a rare cause of acute pancreatitis in children. We report a case of acute pancreatitis in which abdominal sonography revealed an enlarged hypoechoic edematous pancreas with mildly dilated main pancreatic duct and a cystic structure with layered wall in the second part of duodenum. Abdominal CT yielded similar findings. The diagnosis of duodenal duplication was confirmed at surgery and subsequent histopathologic examination. PMID- 18393381 TI - Analysis of the gastrin-releasing peptide receptor gene in Italian patients with autism spectrum disorders. AB - The gastrin-releasing peptide receptor (GRPR) was implicated for the first time in the pathogenesis of Autism spectrum disorders (ASD) by Ishikawa-Brush et al. [Ishikawa-Brush et al. (1997): Hum Mol Genet 6: 1241-1250]. Since this original observation, only one association study [Marui et al. (2004): Brain Dev 26: 5-7] has further investigated, though unsuccessfully, the involvement of the GRPR gene in ASD. With the aim of contributing further information to this topic we have sequenced the entire coding region and the intron/exon junctions of the GRPR gene in 149 Italian autistic patients. The results of this study led to the identification of four novel point mutations, two of which, that is, C6S and L181F, involve amino acid changes identified in two patients with ASD and Rett syndrome, respectively. Both the leucine at position 181 and the cysteine at position 6 are strongly conserved in vertebrates. C6S and L181F mutant proteins were expressed in COS-7 and BALB/3T3 cells, but they did not affect either GRP's binding affinity or its potency for stimulating phospholipase C-mediated production of inositol 1,4,5-trisphosphate. In summary, our results do not provide support for a major role of the GRPR gene in ASD in the population of patients we have studied. However, there is a potential role of C6S and L181F mutations on GRPR function, and possibly in the pathogenesis of the autistic disorders in the two patients. PMID- 18393382 TI - Neurturin suppresses injury-induced neuronal activating transcription factor 3 expression in cultured guinea pig cardiac ganglia. AB - Cultured guinea pig atrial whole mounts containing the intrinsic cardiac ganglia were used as an in vitro model to investigate the induction of the stress/injury marker activating transcription factor 3 (ATF-3). ATF-3 expression was quantified by using immunocytochemical labeling and real-time PCR. In freshly isolated ganglia, no neuronal or Schwann cell nuclei exhibited ATF-3 immunoreactivity. In 2-hour cultures, the induction of ATF-3 expression was evident in many Schwann cell nuclei, whereas no neuronal nuclei were ATF-3 immunoreactive. Beginning at 4 hours, the percentage of neurons with ATF-3-immunoreactive nuclei increased progressively, and, by 48 hours in culture, approximately 95% of the cardiac neurons had ATF-3-immunoreactive nuclei. Neurturin significantly suppressed ATF-3 expression in 48-hour-cultured neurons without effect on ATF-3 expression in Schwann cell nuclei. Neuturin also could reverse neuronal ATF-3 expression after its induction. The suppression of ATF-3 induction by neurturin was mediated by activation of the phosphatidylinositol 3-kinase and mitogen-activated protein kinase pathways. Glial-derived neurotrophic factor (GDNF) also suppressed neuronal ATF-3 induction during culture. However, culture in serum-free media, presence of nerve growth factor, or addition of pituitary adenylate cyclase activating polypeptide had no effect on ATF-3 induction in the 48-hour-cultured cardiac neurons. By 4 hours in culture, there was a significant increase in ATF-3 transcript levels, and neurturin partially suppressed ATF-3 transcript levels in 48-hour cultures. It is proposed that the loss of target-derived neurturin is a potential mechanism stimulating injury-induced expression of ATF-3 in cardiac neurons. PMID- 18393383 TI - Lack of effect of butylparaben and methylparaben on the reproductive system in male rats. AB - BACKGROUND: Parabens are widely used preservatives in cosmetics and pharmaceutical products, and approved as food additives. Parabens have been considered safe for these uses for many years. Recently, adverse effects on male reproductive parameters in rats have been reported when parabens were given orally for 8 weeks starting at three weeks of age. Our studies used two representative parabens, methyl- and butylparaben, to try to replicate these studies and thereby evaluate potential reproductive effects in male Wistar rats. METHODS: Diets containing 0, 100, 1000 or 10,000 ppm of either butyl- or methylparaben were fed to male rats for eight weeks. Rats were 22 days of age at the start of exposure. Parameters evaluated included organ weights, histopathology of reproductive tissues, sperm production, motility, morphology and reproductive hormone levels (butylparaben only). RESULTS: None of the parameters evaluated for either paraben showed compound- or dosage-dependent adverse effects. Metabolism experiments of butylparaben indicate that it is rapidly metabolized by non-specific esterases to p-hydroxybenzoic acid and butanol, neither of which is estrogenic. CONCLUSIONS: Exposure to methyl- or butylparaben in the diet for eight weeks did not affect any male reproductive organs or parameters at exposures as high as 10,000 ppm, corresponding to a mean daily dose of 1,141.1+/-58.9 or 1,087.6+/-67.8 mg/kg/day for methyl- and butylparaben, respectively. The rapid metabolism of parabens by esterases probably explains why these weakly estrogenic substances elicit no in vivo effects when administered by relevant exposure routes (i.e., topical and oral). PMID- 18393384 TI - The Arabidopsis class VIII myosin ATM2 is involved in endocytosis. AB - Members of the class XI of the myosin superfamily comprising higher plant, actin based molecular motors have been shown to be involved in peroxisome and Golgi vesicle trafficking comparable to yeast and animal class V myosins. The tasks of the second class of myosins of higher plants, class VIII, are unclear. In this study the class VIII myosin ATM2 from the model plant Arabidopsis thaliana was selected for the examination of cargo specificity in vivo. Fluorescent protein fusion plasmid constructs with fragments of the ATM2 cDNA were generated and used for Agrobacterium tumefaciens-based transient transformation of Nicotiana benthamiana leaves. The resulting subcellular localization patterns were recorded by live imaging with confocal laser scanning microscopy (CLSM) in epidermal leaf cells. Expression of a nearly full-length construct displayed labeling of filaments and vesicles, a head + neck fragment led to decoration of filaments only. However, expression of fluorescent protein-tagged C-terminal tail domain constructs labeled vesicular structures of different appearance. Most importantly, coexpression of different RFP/YFP-ATM2 tail fusion proteins showed colocalization and, hence, binding to the same type of vesicular target. Further coexpression experiments of RFP/YFP-ATM2 tail fusion proteins with the endosomal marker FYVE and the endosomal tracer FM4-64 demonstrated colocalization with endosomes. Colocalization was also detected by expression of the CFP-tagged membrane receptor BRI1 as marker, which is constantly recycled via endosomes. Occasionally the ATM2 tail targeted to sites at the plasma membrane closely resembling the pattern obtained upon expression of the YFP-ATM1 C-terminal tail. ATM1 is known for its localization at the plasma membrane at sites of plasmodesmata. PMID- 18393385 TI - Immunologic features of HIV-1-infected women on HAART at delivery. AB - BACKGROUND: The conjoint effect of HIV infection and pregnancy on the immune system of women submitted to the prophylactic antiretroviral therapy presently recommended is still poorly understood. METHODS: We evaluated 44 HIV-infected women (HIV) and 45 HIV-negative women (CT) at parturition and we compared them to 20 healthy nonpregnant women (NP). Immunophenotyping of lymphocytes was done by four-color flow cytometry. RESULTS: All HIV-infected women received HAART during pregnancy and 56.8% had viral load <50 copies/mL at delivery. CD4+T cells/mm(3) were lower in HIV (447) than CT (593) and NP (738) (P < 0.05). CD8+T cells/mm(3) were higher in HIV (799) than CT (384) and NP (395) (P < 0.05). NK cells/mm(3) were lower in HIV (146) than in CT (253) and NP (198) (P < 0.05). CD38 expression on CD4+T and on CD8+T cells was higher in HIV (CD4:12.1; CD8:14.9) than in CT(CD4:9.2; CD8:10.2) and NP(CD4:8.6; CD8:6.0) (P < 0.05). However, CD56 expression on CD8+T cells (a marker of cytolytic effector function) was lower in HIV(7%) than in CT(12%) and NP(9%) (P < 0.05). CONCLUSIONS: Even with low levels of viremia, HIV-infected women at delivery showed a different immunologic profile from both healthy non-HIV-infected women in the puerperium and nonpregnant women, with lower CD4+T and higher CD8+T cells, high levels of CD38 expression, but low CD56 expression on CD8+T cells and low NK cell numbers. PMID- 18393386 TI - Beta-catenin deletion in hepatoblasts disrupts hepatic morphogenesis and survival during mouse development. AB - Beta-catenin, the central component of the canonical Wnt pathway, plays important roles in the processes of liver regeneration, growth, and cancer. Previously, we identified temporal expression of beta-catenin during liver development. Here, we characterize the hepatic phenotype, resulting from the successful deletion of beta-catenin in the developing hepatoblasts utilizing Foxa3-cyclization recombination and floxed-beta-catenin (exons 2 through 6) transgenic mice. Beta catenin loss in developing livers resulted in significantly underdeveloped livers after embryonic day 12 (E12) with lethality occurring at around E17 stages. Histology revealed an overall deficient hepatocyte compartment due to (1) increased cell death due to oxidative stress and apoptosis, and (2) diminished expansion secondary to decreased cyclin-D1 and impaired proliferation. Also, the remnant hepatocytes demonstrated an immature phenotype as indicated by high nuclear to cytoplasmic ratio, poor cell polarity, absent glycogen, and decreased expression of key liver-enriched transcription factors: CCAAT-enhancer binding protein-alpha and hepatocyte nuclear factor-4alpha. A paucity of primitive bile ducts was also observed. While the stem cell assays demonstrated no intrinsic defect in hematopoiesis, distorted hepatic architecture and deficient hepatocyte compartments resulted in defective endothelial cell organization leading to overall fetal pallor. CONCLUSION: Beta-catenin regulates multiple, critical events during the process of hepatic morphogenesis, including hepatoblast maturation, expansion, and survival, making it indispensable to survival. PMID- 18393387 TI - Granulin-epithelin precursor as a therapeutic target for hepatocellular carcinoma. AB - Primary liver cancer, hepatocellular carcinoma (HCC), is the fifth most common cancer and the third leading cancer killer in the world. There is no effective therapeutic option for most HCC patients. A new therapeutic strategy is essential. Granulin-epithelin precursor (GEP, also called progranulin, acrogranin, or PC-derived growth factor) was identified as a potential therapeutic target for HCC from our earlier genome-wide expression profiles. We aimed to conduct a detailed investigation with in vitro and animal experiments. We developed the anti-GEP monoclonal antibody (mAb), and examined its effect on hepatoma cells and normal liver cells in vitro. A nude mice model transplanted with human HCC was used to investigate if anti-GEP mAb can inhibit tumor growth in vivo. We demonstrated that anti-GEP mAb inhibited the growth of hepatoma cells but revealed no significant effect on normal liver cells. In the nude mice model transplanted with human HCC, anti-GEP mAb decreased the serum GEP level and inhibited the growth of established tumors in a dose-dependent manner. The anti GEP mAb reduced tumor cell proliferation via the p44/42 MAPK and Akt pathways, and reduced tumor angiogenesis to deprive the nutrient supply with reduced microvessel density and tumor vascular endothelial growth factor level. CONCLUSION: We have shown that anti-GEP antibody can inhibit HCC growth, providing evidence that GEP is a therapeutic target for HCC treatment. PMID- 18393388 TI - A prospective evaluation of computerized tomographic (CT) scanning as a screening modality for esophageal varices. AB - Patients with cirrhosis require endoscopic screening for large esophageal varices. The aims of this study were to determine the cost-effectiveness and patient preferences of a strategy employing abdominal computerized tomography (CT) as the initial screening test for identifying large esophageal varices. In a prospective evaluation,102 patients underwent both CT and endoscopic screening for gastroesophageal varices. Two radiologists read each CT independently; standard upper gastrointestinal endoscopy was the reference standard. Agreement between radiologists, and between endoscopists regarding size of varices was determined using kappa statistic. Cost-effectiveness analysis was performed to determine the optimal screening strategy for varices. Patient preference was assessed by questionnaire. CT was found to have approximately 90% sensitivity in the identification of esophageal varices determined to be large on endoscopy, but only about 50% specificity. The sensitivity of CT in detecting gastric varices was 87%. In addition, a significant number of gastric varices, peri-esophageal varices, and extraluminal pathology were identified by CT that were not identified by endoscopy. Patients overwhelmingly preferred CT over endoscopy. Agreement between radiologists was good regarding the size of varices (Kappa = 0.56), and exceeded agreement between endoscopists (Kappa = 0.36). Use of CT as the initial screening modality for the detection of varices was significantly more cost-effective compared to endoscopy irrespective of the prevalence of large varices. CONCLUSION: Abdominal CT as the initial screening test for varices could be cost-effective. CT also permits evaluation of extra-luminal pathology that impacts management. PMID- 18393389 TI - Caspase activation is associated with spontaneous recovery from acute liver failure. AB - Acute liver failure (ALF) has various causes and is characterized by rapid hepatocyte dysfunction with development of encephalopathy in the absence of preexisting liver disease. Whereas most patients require liver transplantation to prevent the high mortality, some patients recover spontaneously and show complete liver regeneration. Because of the low incidence of ALF, however, the molecular mechanisms of liver dysfunction and regeneration are largely unknown. In this study, we investigated the role of apoptosis and caspases in 70 ALF patients using novel biomarkers that allow the detection of caspase activation in serum samples. Compared with healthy individuals, activation of caspases was strongly enhanced in ALF patients. Interestingly, patients with spontaneous recovery from ALF revealed a significantly higher activation of caspases than patients that required transplantation or died, although in the latter patients extensive DNA fragmentation and signs of nonapoptotic cell death were observed. In the spontaneous survivors, increased caspase activation was accompanied by elevated levels of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6), important cytokines involved in liver regeneration. CONCLUSION: Our data suggest that caspase activation and apoptosis are involved in ALF of patients with spontaneous recovery, whereas caspase-independent cell death might be more relevant in irreversible forms of liver failure. These findings might be important for therapeutic options of ALF but also suggest that measurement of caspase activation might be of prognostic value to predict the outcome of acute liver failure. PMID- 18393390 TI - Differential pattern of lipid droplet-associated proteins and de novo perilipin expression in hepatocyte steatogenesis. AB - Fatty change (steatosis) is the most frequent liver pathology in western countries and is caused by a broad range of disorders such as alcohol abuse and metabolic syndrome. The surface layer of lipid droplets (LDs) contains members of a protein family that share homologous sequences and domains, the so-called PAT proteins, named after their constituents, perilipin, adipophilin, and TIP47. We characterized the LD-associated proteins in normal and diseased liver connected with LD accumulation. Adipophilin and TIP47 are expressed in LDs of vitamin A storing hepatic stellate cells and additionally in LDs of steatotic hepatocytes. Perilipin, which was thought to be characteristic for LDs of adipocytes and steroidogenic cells, becomes de novo expressed in hepatocytes of human steatotic liver. Perilipin splice variant A was found in human steatotic hepatocytes by biochemical, molecular biological, and immunohistochemical methods. Its association with LDs is different from TIP47 and adipophilin, and depends on size and localization of the LDs, suggesting that the different PAT proteins play specific roles during maturation of LDs. PMID- 18393391 TI - Does strong linkage disequilibrium guarantee redundant association results? AB - A substantial amount of effort has been expended recently towards the identification and evaluation of tag single nucleotide polymorphisms; markers that, due to linkage disequilibrium (LD) patterns in the genome, are able to act as "proxies" for other polymorphic sites. As such, these tag markers are assumed to capture, on their own, a large proportion of the genetic variation contributed by a much greater number of polymorphic sites. One important consequence of this is the potential ability to reduce the cost of genotyping in an association study without a corresponding loss of power. This application carries an implicit assumption that strong LD between markers implies high correlation between the accompanying association test results, so that once a tag marker is evaluated for association, its outcome will be representative of all the other markers for which it serves as proxy. We examined this assumption directly. We find that in the null hypothesis situation, where there is no association between the markers and the phenotype, the relationship between LD and the correlation between association test outcomes is clear, though it is not always ideal. In the alternative case, when genetic association does exist in the region, the relationship becomes much more complex. Here, reasonably high LD between markers does not necessarily imply that the association test result of one marker is a direct substitute for that of the other. In these cases, eliminating one of these markers from the set to be genotyped in an association study will lead to a reduction in overall power. PMID- 18393392 TI - High incidence of distal vaginal atresia in mice lacking Tyro3 RTK subfamily. AB - Vaginal atresia is a congenital abnormality of the female genitourinary system, and the specific molecular mechanism leading to failure of vaginal development remains to be elucidated. Here, we report that the female mice lacking Tyro3 RTK subfamily (Tyro3, Axl, and Mer) exhibit a high incidence of distal vaginal atresia. The ratios of the vaginal atresia in Tyro3 RTKs mutant female mice are as follows: 2.5% for Mer(-/-) mice, 4.0% for Axl(-/-)Mer(-/-), 3.7% for Mer(-/ )Tyro3(-/-), 16.06% for Tyro(-/-)Axl(-/-)Mer(-/-) mice. We did not find the vaginal atresia in Axl(-/-), Tyro3(-/-), Axl(-/-) Tyro(-/-), and wild-type mice. These observations suggest that Tyro3 RTKs play roles collaboratively in vaginal development, and Mer is more critical, Axl and Tyro3 support the function of Mer. The phenotype of mice with the vaginal atresia was characterized in this study. Tyro3 RTKs mutant mouse could be a useful model to study the mechanism of vaginal atresia formation. PMID- 18393393 TI - Structure and folding of disulfide-rich miniproteins: insights from molecular dynamics simulations and MM-PBSA free energy calculations. AB - The fold of small disulfide-rich proteins largely relies on two or more disulfide bridges that are main components of the hydrophobic core. Because of the small size of these proteins and their high cystine content, the cysteine connectivity has been difficult to ascertain in some cases, leading to uncertainties and debates in the literature. Here, we use molecular dynamics simulations and MM PBSA free energy calculations to compare similar folds with different disulfide pairings in two disulfide-rich miniprotein families, namely the knottins and the short-chain scorpion toxins, for which the connectivity has been discussed. We first show that the MM-PBSA approach is able to discriminate the correct knotted topology of knottins from the laddered one. Interestingly, a comparison of the free energy components for kalata B1 and MCoTI-II suggests that cyclotides and squash inhibitors, although sharing the same scaffold, are stabilized through different interactions. Application to short-chain scorpion toxins suggests that the conventional cysteine pairing found in many homologous toxins is significantly more stable than the unconventional pairing reported for maurotoxin and for spinoxin. This would mean that native maurotoxin and spinoxin are not at the lowest free energy minimum and might result from kinetically rather than thermodynamically driven oxidative folding processes. For both knottins and toxins, the correct or conventional disulfide connectivities provide lower flexibilities and smaller deviations from the initial conformations. Overall, our work suggests that molecular dynamics simulations and the MM-PBSA approach to estimate free energies are useful tools to analyze and compare disulfide bridge connectivities in miniproteins. PMID- 18393394 TI - The Escherichia coli YdcF binds S-adenosyl-L-methionine and adopts an alpha/beta fold characteristic of nucleotide-utilizing enzymes. PMID- 18393396 TI - Sequence and structural properties of identical mutations with varying phenotypes in human coagulation factor IX. AB - Mutations in human coagulation factor IX cause an X-linked bleeding disorder Hemophilia B, which can be classified as severe, moderately severe and mild based on the plasma levels of factor IX among affected individuals with respect to normal factor IX activity assayed in the patients' plasma (<1%, 2-5%, 6-30%, respectively). Recently, we identified hemophilia B to be a disease with mutations showing clinical variation and speculated that this phenotypic heterogeneity might be a replacement-specific property. Here, we have analyzed the differences in sequence and structural properties among identical mutations with varying phenotypes (IMVPs) by comparing with mutations with uniform phenotypes (MUPs), with recurring reports in Haemophilia B mutation database. Classification of mutations into IMVPs and MUPs has been done based on rigorous systematic evaluation of the clotting activity each mutation is associated with. IMVPs (n = 51) occur in less conserved mutant sites with more tolerated substitutions compared to MUPs (n = 100). A preponderance of CpG site mutations and Arg as the mutated residue in IMVPs compared to Cys in MUPs was observed. Hence, a CpG site substitution at less conserved Arg site might have an increased propensity of expressing variable phenotypes. The changes in intrinsic properties associated with the mutation are less drastic for IMVPs than for MUPs, though no significant differences were observed in structural properties. Based on this study and available literature we speculate that modifier genes at other loci, epigenetic interactions and environment may serve individually or cumulatively to bring about the clinical variation implicating hemophilia B to be deviation from classical Mendelian disorder with complete penetrance. We demonstrate that phenotypic heterogeneity appears to be site-specific also owing to the lesser conservation of the mutant site. PMID- 18393395 TI - MolAxis: efficient and accurate identification of channels in macromolecules. AB - Channels and cavities play important roles in macromolecular functions, serving as access/exit routes for substrates/products, cofactor and drug binding, catalytic sites, and ligand/protein. In addition, channels formed by transmembrane (TM) proteins serve as transporters and ion channels. MolAxis is a new sensitive and fast tool for the identification and classification of channels and cavities of various sizes and shapes in macromolecules. MolAxis constructs corridors, which are pathways that represent probable routes taken by small molecules passing through channels. The outer medial axis of the molecule is the collection of points that have more than one closest atom. It is composed of two dimensional surface patches and can be seen as a skeleton of the complement of the molecule. We have implemented in MolAxis a novel algorithm that uses state-of the-art computational geometry techniques to approximate and scan a useful subset of the outer medial axis, thereby reducing the dimension of the problem and consequently rendering the algorithm extremely efficient. MolAxis is designed to identify channels that connect buried cavities to the outside of macromolecules and to identify TM channels in proteins. We apply MolAxis to enzyme cavities and TM proteins. We further utilize MolAxis to monitor channel dimensions along Molecular Dynamics trajectories of a human Cytochrome P450. MolAxis constructs high quality corridors for snapshots at picosecond time-scale intervals substantiating the gating mechanism in the 2e substrate access channel. We compare our results with previous tools in terms of accuracy, performance and underlying theoretical guarantees of finding the desired pathways. MolAxis is available on line as a web-server and as a stand alone easy-to-use program (http://bioinfo3d.cs.tau.ac.il/MolAxis/). PMID- 18393397 TI - RhoB can adopt a Mg2+ free conformation prior to GEF binding. PMID- 18393399 TI - De novo design of nonpeptidic compounds targeting the interactions between interferon-alpha and its cognate cell surface receptor. AB - Type 1 interferons (IFN) bind specifically to the corresponding receptor, IFNAR. Agonists and antagonists for IFNAR have potential therapeutic value in the treatment of viral infections and systemic lupus erythematosus, respectively. Specific sequences on the surface of IFN, IFN receptor recognition peptides (IRRPs) mediate the binding and signal transduction when IFN interacts with IFNAR. Structural features of two such IRRPs, IRRP-1 and IRRP-3, were used as templates to design small molecule mimetics. In silico screening was used to identify the molecular structural features mimicking their surface characteristics. A set of 26 compounds were synthesized and their ability to interfere with IFN-IFNAR interactions was investigated. Two compounds exhibited antagonist activity, specifically, blocking IFN-inducible Stat phosphorylation Stat complex-DNA binding. Design principles revealed here pave the way toward a novel series of small molecules as antagonists for IFN-IFNAR interactions. PMID- 18393400 TI - Modern natural products chemistry and drug discovery. PMID- 18393401 TI - Design and synthesis of 2- and 3-substituted-3-phenylpropyl analogs of 1-[2 [bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine and 1-[2 (diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine: role of amino, fluoro, hydroxyl, methoxyl, methyl, methylene, and oxo substituents on affinity for the dopamine and serotonin transporters. AB - Novel derivatives of 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3 phenylpropyl)piperazine (GBR 12909, 1) and 1-[2-(diphenylmethoxy)ethyl]-4-(3 phenylpropyl)piperazine (GBR 12935, 2) with various substituents in positions C2 and C3 of the phenylpropyl side chain were synthesized and evaluated for their ability to bind to the dopamine transporter (DAT) and the serotonin transporter (SERT). In the C2 series, the substituent in the S-configuration, with a lone pair of electrons, significantly enhanced the affinity for DAT, whereas the steric effect of the substituent was detrimental to DAT binding affinity. In the C3 series, neither the lone electron pair nor the steric effect of the substituent seemed to affect DAT binding affinity, while sp (2) hybridized substituents had a detrimental effect on affinity for DAT. In the series, the 2 fluoro-substituted (S)-10 had the highest DAT binding affinity and good DAT selectivity, while the 2-amino-substituted (R)-8 showed essentially the same affinity for DAT and SERT. The oxygenated 16 and 18 possessed the best selectivity for DAT. PMID- 18393402 TI - Natural products as leads to potential drugs: an old process or the new hope for drug discovery? PMID- 18393404 TI - New strategies and methods in the discovery of natural product anti-infective agents: the mannopeptimycins. PMID- 18393403 TI - Chemical genetics: exploring the role of the proteasome in cell biology using natural products and other small molecule proteasome inhibitors. PMID- 18393405 TI - Natural products, small molecules, and genetics in tuberculosis drug development. PMID- 18393407 TI - Genome mining for novel natural product discovery. PMID- 18393406 TI - Combinatorial biosynthesis of natural products. PMID- 18393409 TI - Synthesis and pharmacological evaluations of sildenafil analogues for treatment of erectile dysfunction. AB - The 5-[2-ethoxy-5-(4-methylpiperazin-1-ylsulfonyl)phenyl]-1-methyl-3-propyl-1,6 dihydro-7 H-pyrazolo[4,3-d]pyrimidin-7-one, sildenafil, is a cGMP-specific phosphodiesterase-5 (PDE5) inhibitor used for penile erectile dysfunction. In the search for more potent and selective PDE5 inhibitors, new sildenafil analogues (6a-v), characterized by the presence on the sulfonyl group in the 5' position of novel N-4-substituted piperazines or ethylenediamine moiety, were prepared by traditional and microwave-assisted synthesis and tested in rabbit isolated aorta and corpus cavernosum. Similarly to sildenafil, several analogues showed IC50 values in the nanomolar range. In the in vitro studies, all the tested compounds caused concentration-dependent relaxations in both rabbit isolated aorta and corpus cavernosum. All sildenafil analogues potentiated the nitric oxide dependent vasodilation in endothelium-intact rabbit aorta. Compound 6f exhibited great pEC50 value in corpus cavernosum, and compounds 6r and 6u in isolated aorta were found as potent as sildenafil for inhibiting PDE5. Because several analogues were significantly more lipophilic than sildenafil, these compounds may offer a new lead for development of new sildenafil analogues. PMID- 18393411 TI - Assessment of the anisotropy in the molecule Mn19 with a high-spin ground state S = 83/2 by 35 GHz electron paramagnetic resonance. AB - 35 GHz electron paramagnetic resonance experiments on a powder sample of the magnetic molecule Mn 19 with a high-spin ground state S = 83/2 are presented. At low temperatures, the data are well described by the simulated spectra for an isolated spin with a zero-field-splitting parameter D = 0.004 cm (-1), which is, in particular, positive. Hence, Mn 19 is not a single-molecule magnet; the previously observed magnetic hysteresis at ultralow temperatures is likely due to intermolecular dipolar interactions. PMID- 18393412 TI - Tuning the coordination sphere around highly luminescent lanthanide complexes. AB - A series of three ligands designed for the formation of water-soluble luminescent lanthanide complexes is described. All ligands are based on a 6''-carboxy 2,2':6',2''-terpyridine framework linked via a methylene bridge to n-butylamine. The second negatively charged arm consists of a 6-carboxy-2-methylenepyridine for L1, a 6'-carboxy-6-methylene-2,2'-bipyridine for L2, and a 6''-carboxy-6 methylene-2,2':6',2''-terpyridine for L3. The photophysical properties of the Eu and Tb complexes were studied in aqueous solutions by means of absorption spectroscopy and steady-state and time-resolved luminescence spectroscopy. Luminescence excited-state lifetimes were recorded and led to the determination of two water molecules in the first coordination sphere. The europium complexes were characterized by means of (1)H NMR spectroscopy in D 2O and DFT calculations performed at the B3LYP level both in vacuo and in aqueous solution. Finally, the influence of different phosphorylated anions such as HPO 4 (2-), ATP (4-), ADP (3 ), and AMP (2-) on the luminescence properties of the [Eu L X (H 2O) 2] (+) complexes ( X = 1-3) was investigated in buffered aqueous solutions (0.01 M TRIS, pH 7.0), showing a significant interaction of ATP (4-) with [Eu( L2)(H 2O) 2] (+). The coordination of anions was understood in terms of partial decomplexation of one arm of the ligands and water displacement, with formation of ternary species, and it was rationalized on the basis of the structural models of the complexes obtained from DFT calculations. PMID- 18393413 TI - Enthalpy versus entropy in chlorocarbene/alkene addition reactions. AB - We report the first measured activation parameters for the additions of CCl2 and CClF to simple alkenes and demonstrate the existence of enthalpic barriers for CCl2 additions to cyclohexene and 1-hexene. With these two alkenes, additions of PhCCl are "dominated" by entropic contributions to deltaG++ and additions of CCl2 display comparable contributions of deltaH++ and deltaS++, while CClF additions feature dominant enthalpic contributions to deltaG++. Entropic factors, however, control the additions of all three carbenes to the highly reactive alkene, tetramethylethylene. Parallel computational studies are also reported. PMID- 18393414 TI - Self-assembled zinc chlorin rod antennae powered by peripheral light-harvesting chromophores. AB - The multichromophoric dyads 1, 2 and triad 3 have been synthesized by coupling of the appropriately functionalized chlorin derivative with naphthalene diimide dyes through esterification, and subsequent metalation of the chlorin center with zinc acetate. The self-assembly properties of naphthalene diimide (NDI)-zinc chlorin (ZnChl) dyads 1, 2 and triad 3 have been studied in nonpolar, aprotic solvents by UV-vis, CD, and steady-state emission spectroscopy, revealing formation of rod like structures by noncovalent interactions of zinc chlorin units, while the appended naphthalene diimide dyes do not aggregate at the periphery of the rod antennae. In all these systems, photoexcitation of the enveloping naphthalene diimides at 540 and 620 nm, respectively, leads to highly efficient energy transfer processes (FRET; phiET > or = 0.99) to the inner zinc chlorin backbone, as explored by time-resolved fluorescence spectroscopy on the picosecond time scale. The efficiencies of zinc chlorin rod aggregates for the harvesting of solar light are markedly increased from 26% for dyad 2 up to 63% for triad 3, compared to the LH capacity of the monochromophoric aggregates of model system ZnChl 6a. Thus, with the self-assembled zinc chlorin rod antenna based on triad 3, a highly efficient artificial LH system has been achieved. PMID- 18393415 TI - Long-range influence of carbohydrates on the solvation dynamics of water--answers from terahertz absorption measurements and molecular modeling simulations. AB - We present new terahertz (THz) spectroscopic measurements of solvated sugars and compare the effect of two disaccharides (trehalose and lactose) and one monosaccharide (glucose) with respect to the solute-induced changes in the sub picosecond network dynamics of the hydration water. We found that the solute affects the fast collective network motions of the solvent, even beyond the first solvation layer. For all three carbohydrates, we find an increase of 2-4% in the THz absorption coefficient of the hydration water in comparison to bulk water. Concentration-dependent changes in the THz absorption between 2.1 and 2.8 THz of the solute-water mixture were measured with a precision better than 1% and were used to deduce a dynamical hydration shell, which extends from the surface up to 5.7 +/- 0.4 and 6.5 +/- 0.9 A for the disaccharides lactose and trehalose, respectively, and 3.7 +/- 0.9 A for the glucose. This exceeds the values for the static hydration shell as determined, for example, by scattering, where the long range structure was found to be not significantly affected by the solute beyond the first hydration shell. When comparing all three carbohydrates, we found that the solute-induced change in the THz absorption depends on the product of molar concentration of the solute and the number of hydrogen bonds between the carbohydrate and water molecules. We can conclude that the long-range influence on the sub-picosecond collective water network motions of the hydration water is directly correlated with the average number of hydrogen bonds between the molecule and adjacent water molecules for carbohydrates. This implies that monosaccharides have a smaller influence on the surrounding water molecules than disaccharides. This could explain the bioprotection mechanism of sugar-water mixtures, which has been found to be more effective for disaccharides than for monosaccharides. PMID- 18393416 TI - Asymmetric catalysis via dynamic substrate/ligand/rare earth metal conglomerate. AB - A highly enantio- and diastereoselective catalytic asymmetric Mannich-type reaction of alpha-cyanoketones and N-Boc imines promoted by an amide ligand/Sc(OiPr)3 catalyst is described. The similar reaction outcome is obtained with/without precomplexation of catalyst, suggesting that reaction components in a non-ordered conglomerate mixture orchestrate to form an ordered transition state during the reaction. Spectroscopic data and Eyring plot are consistent with this assumption. PMID- 18393417 TI - Selective linear triene formation from different alkynes using Zr/Cu system. AB - Selective synthesis of linear trienes from three different alkynes was achieved in one-pot procedure using the Zr/Cu system. Zirconacyclopentadiene prepared from two different alkynes such as an alkyl-substituted alkyne and an aryl-substituted alkyne reacted with NCS gave chlorodienylzirconocene. It reacted with the third alkyne with electron-withdrawing groups in the presence of CuCl to afford the corresponding linear triene. PMID- 18393418 TI - Toward the characterization of peptidoglycan structure and protein-peptidoglycan interactions by solid-state NMR spectroscopy. AB - Solid-state NMR spectroscopy is applied to intact peptidoglycan sacculi of the Gram-negative bacterium Escherichia coli. High-quality solid-state NMR spectra allow atom-resolved investigation of the peptidoglycan structure and dynamics as well as the study of protein-peptidoglycan interactions. PMID- 18393419 TI - Upconversion multicolor fine-tuning: visible to near-infrared emission from lanthanide-doped NaYF4 nanoparticles. AB - A general approach to fine-tuning the upconversion emission colors, based upon a single host source of NaYF4 nanoparticles doped with Yb3+, Tm3+, and Er3+, is presented. The emission intensity balance can be precisely controlled using different host-activator systems and dopant concentrations. The approach allows access to a wide range of luminescence emission from visible to near-infrared by single-wavelength excitation. PMID- 18393420 TI - Colloidal GaAs quantum wires: solution-liquid-solid synthesis and quantum confinement studies. AB - Colloidal GaAs quantum wires with diameters of 5-11 nm and narrow diameter distributions (standard deviation = 12-21% of the mean diameter) are grown by two methods based on the solution-liquid-solid (SLS) mechanism. Resolved excitonic absorption features arising from GaAs quantum wires are detected, allowing extraction of the size-dependent effective band gaps of the wires. The results allow the first systematic comparison of the size dependences of the effective band gaps in corresponding sets of semiconductor quantum wires and quantum wells. The GaAs quantum wire and well band gaps scale according to the prediction of a simple effective-mass-approximation, particle-in-a-box (EMA-PIB) model, which estimates the kinetic confinement energies of electron-hole pairs in quantum nanostructures of different shapes and confinement dimensionalities. PMID- 18393421 TI - Suggestion of an organometallic intermediate in an intramolecular dechlorination reaction involving copper(I) and a ArCH2Cl moiety. AB - A copper(I)-mediated reductive dechlorination reaction involving an "internal" chloromethylene substrate at the pyridyl 6-position of one TMPA arm (TMPA triple bond TPA triple bond tris(2-pyridylmethyl)amine) leads to a 1:1 ratio of the starting ligand 6ClCH2-TMPA and a new methyl-TMPA product, 6CH2H-TMPA. On the basis of observed product distributions and a kinetic study, a reaction mechanism involving intramolecular oxidative insertion of Cu(I) to the C-Cl substrate is suggested. The resulting organometallic intermediate is then protonated, leading to the observed products. PMID- 18393422 TI - Quantum dot FRET biosensors that respond to pH, to proteolytic or nucleolytic cleavage, to DNA synthesis, or to a multiplexing combination. AB - Fluorescent acceptors have been immobilized on nanoparticulate quantum dots (QDs), which serve in turn as their FRET donors. The broad excitation and narrow emission bands of QDs mark them as having excellent potential as donors for FRET and, in principle, differently colored QDs could be excited simultaneously. The present work describes the preparation and operation of FRET-based QD bioprobes individually able to detect the actions of protease, deoxyribonuclease, DNA polymerase, or changes in pH. In addition, two such QD-mounted biosensors were excited at a single wavelength, and shown to operate simultaneously and independently of each other in the same sample solution, allowing multiplex detection of the action of a protease, trypsin, in the presence of deoxyribonuclease. PMID- 18393423 TI - Toward three-dimensional nanoengineering of heterogeneous catalysts. AB - Cobalt-based Fischer-Tropsch systems are widely used to convert synthesis gas to clean hydrocarbon fuel. However, surprisingly little is known about the morphology of the catalysts on the nanoscale. Here we show that scanning transmission electron tomography reveals their true 3-D morphology and provides direct evidence that the support controls the final morphology of the catalyst. Such direct local three-dimensional measurements provide unprecedented insight into catalysis, and can henceforth transform our understanding of these complex materials. PMID- 18393424 TI - Transition-state analysis of the DNA repair enzyme MutY. AB - The transition state (TS) structure of MutY-catalyzed DNA hydrolysis was solved using multiple kinetic isotope effect (KIE) measurements. MutY is a base excision repair enzyme which cleaves adenine from 8-oxo-G:A mismatches in vivo, and also from G:A mismatches in vitro. TS analysis of G:A-DNA hydrolysis revealed a stepwise S(N)1 (D(N)*A(N)(double dagger)) mechanism proceeding through a highly reactive oxacarbenium ion intermediate which would have a lifetime in solution of <10(-10) s. C-N bond cleavage is reversible; the N-glycoside bond breaks and reforms repeatedly before irreversible water attack on the oxacarbenium ion. KIEs demonstrated that MutY uses general acid catalysis by protonating N7. It enforces a 3'-exo sugar ring conformation and other sugar ring distortions to stabilize the oxacarbenium ion. Combining the experimental TS structure with the previously reported crystal structure of an abortive Michaelis complex elucidates the step by-step catalytic sequence. PMID- 18393425 TI - STM studies of fusion of cholesterol suspensions and mixed 1,2-dimyristoyl-sn glycero-3-phosphocholine (DMPC)/cholesterol vesicles onto a Au(111) electrode surface. AB - Electrochemical scanning tunneling microscopy (EC-STM) has been applied to study the structure of the film formed by fusion of cholesterol suspensions and mixed dimyristoylphosphatidylcholine (DMPC)/cholesterol vesicles on a Au(111) electrode surface. It has been demonstrated that cholesterol molecules assemble at the gold support into several structures templated by the crystallography of the metal surface and involving flat or edge-on adsorbed molecules. Studies of the film formed by fusion of mixed DMPC/cholesterol vesicles revealed that ordered domains of either pure DMPC or pure cholesterol were formed. These results indicate that, at the metal surface, the molecules released by the rupture of a vesicle initially self-assemble into a well-ordered monolayer. The self-assembly is controlled by the hydrocarbon skeleton-metal surface interaction. In the case of mixed DMPC/cholesterol vesicles, the molecule-metal interactions induce segregation of the two components into single component domains. However, the molecule-metal interaction induced monolayer is a transient phenomenon. When more molecules accumulate at the surface, the molecule-molecule interactions dominate the assembly, and the monolayer is transformed into a bilayer. PMID- 18393426 TI - Reaction of molecular oxygen with a Pd(II)-hydride to produce a Pd(II) hydroperoxide: experimental evidence for an HX-reductive-elimination pathway. AB - The reaction of molecular oxygen with a Pd(II)-hydride species to form a Pd(II) hydroperoxide represents one of the proposed catalyst reoxidation pathways in Pd catalyzed aerobic oxidation reactions, but well-defined examples of this reaction were discovered only recently. Here, we present a mechanistic study of the reaction of O2 with trans-(IMes) 2Pd(H)(OBz), 1 (IMes = 1,3-dimesitylimidazol-2 ylidene), which yields trans-(IMes) 2Pd(OOH)(OBz), 2. The reaction was monitored by (1)H NMR spectroscopy in benzene-d6, and kinetic studies reveal a two-term rate law, rate = k1[1] + k2[1][BzOH], and a small deuterium kinetic isotope effect, k(Pd-H)/k(Pd-D) = 1.3(1). The rate is independent of the oxygen pressure. The data support a stepwise mechanism for the conversion of 1 into 2 consisting of rate-limiting reductive elimination of BzOH from 1 followed by rapid reaction of molecular oxygen with (IMes) 2Pd(0) and protonolysis of a Pd-O bond of the eta(2)-peroxo complex (IMes) 2Pd(O2). Benzoic acid and other protic additives (H2O, ArOH) catalyze the oxygenation reaction, probably by stabilizing the transition state for reductive elimination of BzOH from 1. This study provides the first experimental validation of the mechanism traditionally proposed for aerobic oxidation of Pd-hydride species. PMID- 18393427 TI - Identification of acidic phosphorus-containing ligands involved in the surface chemistry of CdSe nanoparticles prepared in tri-N-octylphosphine oxide solvents. AB - The surface ligand composition of CdSe nanoparticles prepared using technical grade tri-n-octylphosphine oxide (TOPO) was investigated using a nucleophilic ligand displacement methodology and (31)P {(1)H} NMR spectroscopy. 4-(N,N Dimethylamino)pyridine (DMAP) and benzyltrimethylammonium propionate were added to tetrahydrofuran solutions of CdSe nanoparticles prepared in technical grade TOPO. DMAP was shown to be a sufficiently strong nucleophile to displace the more weakly coordinating ligands, TOPO, TOPSe, di-n-octylphosphinate, and n octylphosphonate (OPA). Benzyltrimethylammonium propionate was shown to be a stronger nucleophile than DMAP in that it could displace all the aforementioned surface-bound ligands as well as a previously unidentified surface-bound phosphorus species. Independent synthesis and (31)P {(1)H} NMR spectral matching confirmed that the new species was P,P'-(di-n-octyl) dihydrogen pyrophosphonic acid (PPA). The PPA was shown to form during the nanoparticle synthesis via the dehydrative condensation of OPA. CdSe nanoparticle syntheses were performed using pure TOPO and added OPA, and subsequent displacement experiments showed that OPA and PPA were the predominant surface-bound ligands. CdSe nanoparticle syntheses were performed using pure TOPO and added PPA, and subsequent displacement experiments showed that PPA was the predominant surface-bound ligand. PPA was also shown to have the greatest affinity for the nanoparticle surface of all the ligands investigated. Thus, a model for the surface ligand composition could be developed for nanoparticles prepared using technical grade TOPO or other high boiling solvents with added acidic phosphorus compounds. PMID- 18393428 TI - Effect of the seasoning method on the chemical composition of oak heartwood to cooperage. AB - The effect of various oak wood seasoning methods, natural seasoning in open air, artificial seasoning in a kiln, and a mixed method that combines open air and kiln drying, on the chemical composition in American ( Quercus alba) and French ( Quercus petraea) oak was studied. The results confirm the evolution of ellagitannins in wood, low molecular polyphenols, and volatile compounds in relation to oak species and seasoning method. Natural seasoning in open air can be considered a superior method to mixed or artificial drying for wood seasoning in cooperage. It was more effective in reducing the excess of ellagitannins, especially in French oak wood. Moreover, the evolution of wood volatile compounds was more positive in the natural seasoning than mixed or artificial drying, since it led to woods with higher aromatic potential (larger concentrations of compounds such as volatile phenols, phenolic aldehydes, furanic compounds, and cis- and trans-beta-methyl-gamma-octalactones) than mixed or artificial methods. However, the three seasoning methods showed similar effectiveness regarding the desired reduction of off-flavors compounds. PMID- 18393429 TI - Rapid enzyme-linked immunosorbent assay and colloidal gold immunoassay for kanamycin and tobramycin in Swine tissues. AB - A monoclonal antibody (Mab) was produced by using the kanamycin-glutaraldehyde bovine serum albumin (Kan-GDA-BSA) conjugate as the immunogen. The anti-Kan Mab exhibited high cross-reactivity with tobramycin (Tob) and slight or negligible cross-reactivity with other aminoglycosides. The specificity and cross-reactivity of this Mab are discussed regarding the three-dimensional, computer-generated molecular models of the aminoglycosides. Using this Mab, a rapid enzyme-linked immunosorbent assay (ELISA) and a colloidal gold-based strip test for Kan and Tob were developed. The rapid ELISA showed a 50% inhibition value (IC 50) of 0.83 ng/mL for Kan and 0.89 ng/mL for Tob with the analysis time less than 40 min, and the recoveries from spiked swine tissues at levels of 25-200 microg/kg ranged from 52% to 96% for Kan and 61% to 86% for Tob. In contrast, the strip test for Kan or Tob had a visual detection limit of 5 ng/mL in PBS, 50 microg/kg in meat or liver, and 100 microg/kg in kidney, and the results can be judged within 5-10 min. Observed positive samples judged by the strip test can be further quantitated by ELISA, hence the two assays can complement each other for rapid detection of residual Kan and Tob in swine tissues. Moreover, physical-chemical factors that affected the ELISA and strip test performance were also investigated. The effect of pH and antibody amount for gold-antibody conjugation on the strip test sensitivity was determined followed by a theoretical explanation of the effects. PMID- 18393430 TI - Irrigation with diluted seawater improves the nutritional value of cherry tomatoes. AB - The aim of this study was to assess whether the nutritional value of cherry tomato can be improved by irrigating plants with diluted seawater (12%; EC = 10 mS/cm in comparison with a control at EC = 4 mS/cm). Berries of cherry tomato cv. Naomi were analyzed at the red-ripe stage for the contents of NADPH and NADP+ as well as for the amounts of the main antioxidants, such as ascorbic acid, lipoic acid, tocopherols, and phenolic acids. As compared to the controls, the fruits of salt-treated plants showed a higher titratable acidity and a higher concentration of reducing sugars. The fruits picked from tomato plants irrigated with diluted seawater produced berries characterized by a higher nutritional value. Following salinity, berries showed higher amounts of vitamin C, vitamin E, dihydrolipoic acid, and chlorogenic acid. It was hypothesized that protocatechuic, vanillic, caffeic, and ferulic acids were utilized to counteract the damaging effects of salinity-induced oxidative stress, allowing tomato fruits to maintain a high reduced status even following salinization. PMID- 18393431 TI - Effects of the mango components mangiferin and quercetin and the putative mangiferin metabolite norathyriol on the transactivation of peroxisome proliferator-activated receptor isoforms. AB - Mangos are a source of bioactive compounds with potential health-promoting activity. This study evaluated the abilities of the mango components quercetin and mangiferin and the aglycone derivative of mangiferin, norathyriol, to modulate the transactivation of peroxisome proliferator-activated receptor isoforms (PPARs). PPARs are transcription factors important in many human diseases. Through the use of a gene reporter assay it was shown that quercetin inhibited the activation of all three isoforms of PPARs (PPARgamma IC(50) = 56.3 microM; PPARalpha IC(50) = 59.6 microM; PPARbeta IC(50) = 76.9 microM) as did norathyriol (PPARgamma IC(50) = 153.5 microM; PPARalpha IC(50) = 92.8 microM; PPARbeta IC(50) = 102.4 microM), whereas mangiferin did not inhibit the transactivation of any isoform. These findings suggest that mango components and metabolites may alter transcription and could contribute to positive health benefits via this or similar mechanisms. PMID- 18393432 TI - Simple assessment of radical scavenging capacity of beverages. AB - The radical-scavenging antioxidants play an important role against oxidative stress in the defense system in vivo. The beneficial effects of antioxidants contained in foods and beverages have been well-accepted, and their antioxidant capacity has been assessed by various methods. In the present study, a simple method is proposed in which the total radical scavenging capacity is assessed from the bleaching of pyranine and pyrogallol red induced by free radicals generated from azo initiator. The total content of antioxidants contained in red wine, green tea, and cassis drink and their reactivities toward peroxyl radicals were measured from the lag phase and rate of bleaching using pyranine and pyrogallol red as a probe, respectively. It was found that this method to follow the bleaching of two probes by visible light spectrophotometer is convenient and applicable for assessment of total radical scavenging capacity of both content and activity of the antioxidants contained in beverages. PMID- 18393433 TI - Preparation and phytotoxicity of novel kaurane diterpene amides with potential use as herbicides. AB - Novel kaurane ditepene monoamides were synthesized in good yields directly from kaurenoic ( 1) and grandiflorenic ( 2) acids and unprotected symmetrical diamines, using a modified protocol for monoacylation. Amides from 1 and 2 and monoamines were also obtained and tested against seed germination and growth of radicle and shoot of Lactuca sativa (lettuce), at 10 (-3), 10 (-5), and 10 (-7) M. Amides from symmetrical diamines showed significant inhibitory activity at higher concentrations. PMID- 18393434 TI - Characterization of odor-active compounds in extracts obtained by simultaneous extraction/distillation from moroccan black olives. AB - "Greek-style" Moroccan black table olives were screened for potent odorants by GC/olfactometry/aroma extract dilution analysis of representative Likens Nickerson extracts and compared with "Spanish-style" green fruits. ( Z)-3 Hexenal, ( E, E)-2,4-decadienal, ( E, Z)-2,4-decadienal, guaiacol, and methional were found in both green and black olives, but with significant differences in concentration according to the fruit ripening degree (the first was lower and the last two were higher in black fruits). Specific compounds not previously detected in green olives (gamma-deca- and dodecalactones, delta-decalactone, and 2-methyl 3-furanthiol) proved to be, with methional, the strongest odors in black olive extracts. These extracts were also distinguishable from green olive extracts by the presence of new sulfur compounds and fewer terpenes. PMID- 18393435 TI - Identification of fructooligosaccharides in different banana cultivars. AB - Banana has been currently indicated as a good source of fructooligosaccharides (FOS), which are considered to be functional components of foods. However, significant differences in their amounts in bananas have been observed in the literature. This work aims to identify and quantify FOS during ripening in different banana cultivars belonging to the most common genomic groups cultivated in Brazil. Considering that these differences can be due to cultivar, stage of ripening, and the methodologies used for FOS analyses, sugar contents were analyzed by high performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD) and gas chromatography-mass spectrometry (GC-MS). An initial screening of eight cultivars (Ouro, Nanicao, Prata, Maca, Mysore, Pacovan, Terra, and Figo) in a full-ripe stage showed that 1-kestose, the first member of the FOS series (amounts between 297 and 1600 microg/g of DM), was accumulated in all of them. Nystose, the second member, was detected only in Prata cultivar. Five of the cultivars were analyzed during ripening, and a strong correlation could be established with a specific sucrose level ( approximately 200 mg/g of DM), which seems to trigger the synthesis of 1-kestose (the low amounts of FOS, below the functional recommended dose, indicates that banana cannot be considered a good source of FOS). PMID- 18393436 TI - Prediction of atrazine sorption coefficients in soils using mid-infrared spectroscopy and partial least-squares analysis. AB - This study explored the potential of mid-infrared spectroscopy (MIR) with partial least-squares (PLS) analysis to predict sorption coefficients (Kd) of pesticides in soil. The MIR technique has the advantage of being sensitive to both the content and the chemistry of soil organic matter and mineralogy, the important factors in the sorption of nonionic pesticides. MIR spectra and batch Kd values of atrazine were determined on a set of 31 soil samples as reference data for PLS calibration. The samples, with high variability in soil organic carbon content (SOC), were chosen from 10 southern Australian soil profiles (A1, A2, B, and C in one case). PLS calibrations, developed for the prediction of Kd from the MIR spectra and reference Kd data, were compared with predictions from Koc-based indirect estimation using SOC content. The reference Kd data for the 31 samples ranged from 0.31 to 5.48 L/kg, whereas Koc ranged from 30 to 680 L/kg. Both coefficients generally increased with total SOC content but showed a relatively poor coefficient of determination (R2 = 0.53; P > 0.0001) and a high standard error of prediction (SEP =1.22) for the prediction of Kd from Koc. This poor prediction suggested that total SOC content alone could explain only half of the variation in Kd. In contrast, the regression plot of PLS predicted versus measured Kd resulted in an improved correlation, with R2 = 0.72 ( P > 0.0001) and standard error of cross-validation (SECV) = 0.63 for three PLS factors. With the advantages of MIR-PLS in mind, (i) more accurate prediction of Kd, (ii) an ability to reflect the nature and content of SOC as well as mineralogy, and (iii) high repeatability and throughput, it is proposed that MIR-PLS has the potential for an improved and rapid assessment of pesticide sorption in soils. PMID- 18393437 TI - Antimicrobial, dehydroascorbate reductase, and monodehydroascorbate reductase activities of defensin from sweet potato [Ipomoea batatas (L.) Lam. 'Tainong 57'] storage roots. AB - A cDNA encoding a small cysteine-rich protein designated defensin (SPD1) was isolated from sweet potato storage roots. On the basis of the amino acid sequence similarity and conserved residues, it is suggested that SPD1 is a member of the plant defensin family. Recombinant SPD1 protein overproduced in Escherichia coli was purified by Ni (2+)-chelated affinity chromatography. A recombinant protein from the storage root cDNA clone effectively inhibited the trypsin activity in a dose-dependent manner. Both the corresponding mRNA and protein level were found to be highest in the storage roots, followed by sprout. SPD1 reduced dehydroascorbate (DHA) in the presence of glutathione to regenerate l-ascorbic acid (AsA). However, without glutathione, SPD1 has very low DHA reductase activity, and AsA was oxidized by AsA oxidase to generate monodehydroascorbate (MDA) free radical. MDA was also reduced by SPD1 to AsA in the presence of NADH, mimicking the MDA reductase catalyzed reaction. These data suggest that SPD1 has both DHA reductase and MDA reductase activities. SPD1 was also shown to inhibit the growth of both fungi and bacteria. SPD1 is apparently the first reported plant defensin exhibiting DHA and MDA activities in vitro. PMID- 18393438 TI - The impact of molecular weight and PEG chain length on the systemic pharmacokinetics of PEGylated poly l-lysine dendrimers. AB - The impact of PEGylation on the pharmacokinetics and biodistribution of (3)H labeled poly l-lysine dendrimers has been investigated after intravenous administration to rats. The volumes of distribution, clearance and consequently the plasma half-lives of the PEGylated dendrimers were markedly dependent on the total molecular weight of the PEGylated dendrimer, but were not specifically affected by the PEG chain length alone. In general, the larger dendrimer constructs (i.e. >30 kDa) had reduced volumes of distribution, were poorly renally cleared and exhibited extended elimination half-lives ( t 1/2 1-3 days) when compared to the smaller dendrimers (i.e. <20 kDa) which were rapidly cleared from the plasma principally into the urine ( t 1/2 1-10 h). At later time points the larger dendrimers concentrated in the organs of the reticuloendothelial system (liver and spleen); however, the absolute extent of accumulation was low. Size exclusion chromatography of plasma and urine samples revealed that the PEGylated dendrimers were considerably more resistant to biodegradation in vivo than the underivatized poly l-lysine dendrimer cores. The results suggest that the size of PEGylated poly l-lysine dendrimer complexes can be manipulated to optimally dictate their pharmacokinetics, biodegradation and bioresorption behavior. PMID- 18393439 TI - Platelet endothelial cell adhesion molecule 1 (PECAM-1) and its interactions with glycosaminoglycans: 1. Molecular modeling studies. AB - Platelet endothelial cell adhesion molecule 1 (PECAM-1) has many functions, including its roles in leukocyte extravasation as part of the inflammatory response and in the maintenance of vascular integrity through its contribution to endothelial cell-cell adhesion. PECAM-1 has been shown to mediate cell-cell adhesion through homophilic binding events that involve interactions between domain 1 of PECAM-1 molecules on adjacent cells. However, various heterophilic ligands of PECAM-1 have also been proposed. The possible interaction of PECAM-1 with glycosaminoglycans (GAGs) is the focus of this study. The three-dimensional structure of the extracellular immunoglobulin (Ig) domains of PECAM-1 were constructed using homology modeling and threading methods. Potential heparin/heparan sulfate-binding sites were predicted on the basis of their amino acid consensus sequences and a comparison with known structures of sulfate binding proteins. Heparin and other GAG fragments have been docked to investigate the structural determinants of their protein-binding specificity and selectivity. The modeling has predicted two regions in PECAM-1 that appear to bind heparin oligosaccharides. A high-affinity binding site was located in Ig domains 2 and 3, and evidence for a low-affinity site in Ig domains 5 and 6 was obtained. These GAG-binding regions were distinct from regions involved in PECAM-1 homophilic interactions. PMID- 18393440 TI - Inhibitors of the molybdenum cofactor containing 4-hydroxybenzoyl-CoA reductase. AB - 4-Hydroxybenzoyl-CoA reductase (4-HBCR) is a member of the xanthine oxidase (XO) family of molybdenum cofactor containing enzymes and catalyzes the irreversible removal of a phenolic hydroxy group by reduction, yielding benzoyl-CoA and water. In this work the effects of various activity modulating compounds were characterized by kinetic, electron paramagnetic resonance (EPR) spectroscopic, and X-ray crystallographic studies. 4-HBCR was readily inactivated by cyanide and by the reducing agents titanium(III) citrate and dithionite; in contrast, reduced viologens had no inhibitory effect. Cyanide inhibition occurred in both the oxidized and reduced state of 4-HBCR. In the reduced state, cyanide-inhibited 4 HBCR was reactivated by simple oxidation. In contrast, reactivation from the oxidized state was only achieved in the presence of sulfide. Dithionite-inhibited 4-HBCR was reactivated by oxidation, whereas inhibition by titanium(III) citrate was irreversible. The previously reported inhibitory effect of azide could not be confirmed; instead, azide rather protected the enzyme from inactivation by titanium(III) citrate. The EPR spectra of the Mo(V) states were nearly identical in the noninhibited methyl viologen and in the dithionite-inhibited states of 4 HBCR; they exhibited a hyperfine splitting due to magnetic coupling with two solvent-exchangeable protons. The cyanide-treated enzyme showed the typical desulfo-inhibited Mo(V) EPR signal in D 2O, whereas in H 2O the hyperfine splitting was altered but indicated no loss of Mo(V)-proton interactions. The structures of dithionite- and azide-bound 4-HBCR were solved at 2.1 and 2.2 A, respectively. Both dithionite and azide bound directly to equatorial ligation sites of the Mo atom. The results obtained revealed further insights into the active site of an unusual member of the XO family of molybdenum cofactor containing enzymes. PMID- 18393441 TI - Structural basis of the iron storage function of frataxin from single-particle reconstruction of the iron-loaded oligomer. AB - The mitochondrial protein frataxin plays a central role in mitochondrial iron homeostasis, and frataxin deficiency is responsible for Friedreich ataxia, a neurodegenerative and cardiac disease that affects 1 in 40000 children. Here we present a single-particle reconstruction from cryoelectron microscopic images of iron-loaded 24-subunit oligomeric frataxin particles at 13 and 17 A resolution. Computer-aided classification of particle images showed heterogeneity in particle size, which was hypothesized to result from gradual accumulation of iron within the core structure. Thus, two reconstructions were created from two classes of particles with iron cores of different sizes. The reconstructions show the iron core of frataxin for the first time. Compared to the previous reconstruction of iron-free particles from negatively stained images, the higher resolution of the present reconstruction allowed a more reliable analysis of the overall three dimensional structure of the 24-meric assembly. This was done after docking the X ray structure of the frataxin trimer into the EM reconstruction. The structure revealed a close proximity of the suggested ferroxidation sites of different monomers to the site proposed to serve in iron nucleation and mineralization. The model also assigns a new role to the N-terminal helix of frataxin in controlling the channel at the 4-fold axis of the 24-subunit oligomer. The reconstructions show that, together with some common features, frataxin has several unique features which distinguish it from ferritin. These include the overall organization of the oligomers, the way they are stabilized, and the mechanisms of iron core nucleation. PMID- 18393442 TI - A selenocysteine variant of the human copper chaperone for superoxide dismutase. A Se-XAS probe of cluster composition at the domain 3-domain 3 dimer interface. AB - We report the semisynthesis of a selenocysteine (Sec) derivative of the human copper chaperone for superoxide dismutase, substituted with Sec at the C-terminal C246 residue. Measurements of hCCS-induced SOD1 activation were used to show that the C-terminal CXC sequence is both necessary and sufficient for EZn-SOD maturation. Therefore, an active CAU variant carrying Sec as the terminal amino acid was prepared by expressed protein ligation of a single selenocysteine amino acid to a 243-CA truncation. This reaction proceeded in high yield and generated the desired 243-CAX (X = C or U) protein with the expected mass. Se-edge XAS of the apoprotein indicated that both Se-S and Se-Se interactions were present in a 0.3:0.7 ratio, indicating an equilibrium between species with either a selenosulfide or a diselenide cross-link. After reduction on immobilized TCEP, the ligated Cys and Sec apoproteins bound up to 2.5 Cu(I) ions per hCCS monomer with both Cu and Se as constituent atoms of the cluster which forms at the domain 3 interface of a hCCS dimer. Merging of XAS data at the Cu and Se K-absorption edges provided additional details of the cluster composition, specifically the fact that both Se atoms occupied bridging positions between two Cu(I) atoms. Further, the requirement for identical Cu-Se bond lengths and Debye-Waller factors at each absorption edge allowed us to rule out simple models for the cluster composition such as a bis-Cys(Sec)-bridged dinuclear cluster and was indicative of a more complex cluster with a nuclearity of >or=3. PMID- 18393443 TI - Cyanolysis and azidolysis of epoxides by haloalcohol dehalogenase: theoretical study of the reaction mechanism and origins of regioselectivity. AB - Haloalcohol dehalogenase HheC catalyzes the reversible dehalogenation of vicinal haloalcohols to form epoxides and free halides. In addition, HheC is able to catalyze the irreversible and highly regioselective ring-opening of epoxides with nonhalide nucleophiles, such as CN (-) and N 3 (-). For azidolysis of aromatic epoxides, the regioselectivity observed with HheC is opposite to the regioselectivity of the nonenzymatic epoxide-opening. This, together with a relatively broad substrate specificity, makes HheC a promising tool for biocatalytic applications. We have designed large quantum chemical models of the HheC active site and used density functional theory to study the reaction mechanism of the HheC-catalyzed ring-opening of ( R)-styrene oxide with the nucleophiles CN (-) and N 3 (-). Both the cyanolysis and the azidolysis reactions are shown to take place in a single concerted step. The results support the suggested role of the putative Ser132-Tyr145-Arg149 catalytic triad, where Tyr145 acts as a general acid, donating a proton to the substrate, and Arg149 interacts with Tyr145 and facilitates proton abstraction, while Ser132 positions the substrate and reduces the barrier for epoxide opening through interaction with the emerging oxyanion of the substrate. We have also studied the regioselectivity of ( R)-styrene oxide opening for both the cyanolysis and the azidolysis reactions. The employed active site model was shown to be able to reproduce the experimentally observed beta-regioselectivity of HheC. In silico mutations of various groups in the HheC active site model were performed to elucidate the important factors governing the regioselectivity. PMID- 18393444 TI - Cu(II) binding to monomeric, oligomeric, and fibrillar forms of the Alzheimer's disease amyloid-beta peptide. AB - Copper has been proposed to play a role in Alzheimer's disease through interactions with the amyoid-beta (Abeta) peptide. The coordination environment of bound copper as a function of Cu:Abeta stoichiometry and Abeta oligomerization state are particularly contentious. Using low-temperature electron paramagnetic resonance (EPR) spectroscopy, we spectroscopically distinguish two Cu(II) binding sites on both soluble and fibrillar Abeta (for site 1, A parallel = 168 +/- 1 G and g parallel = 2.268; for site 2, A parallel = 157 +/- 2 G and g parallel = 2.303). When fibrils that have been incubated with more than 1 equiv of Cu(II) are washed, the second Cu(II) ion is removed, indicating that it is only weakly bound to the fibrils. No change in the Cu(II) coordination environment is detected by EPR spectroscopy of Cu(II) with Abeta (1:1 ratio) collected as a function of Abeta fibrillization time, which indicates that the Cu(II) environment is independent of Abeta oligomeric state. The initial Cu(II)-Abeta complexes go on to form Cu(II)-containing Abeta fibrils. Transmission electron microscopy images of Abeta fibrils before and after Cu(II) addition are the same, showing that once incorporated, Cu(II) does not affect fibrillar structure; however, the presence of Cu(II) appears to induce fibril-fibril association. On the basis of our results, we propose a model for Cu(II) binding to Abeta during fibrillization that is independent of peptide oligomeric state. PMID- 18393445 TI - Characterization of a lysine-specific histone demethylase from Arabidopsis thaliana. AB - Arabidopsis thaliana has four genes with close homology to human histone H3 lysine 4 demethylase (HsLSD1), a component of various transcriptional corepressor complexes that often also contain histone deacetylases and the corepressor protein CoREST. All four Arabidopsis proteins contain a flavin amine oxidase domain and a SWIRM domain, the latter being present in a number of proteins involved in chromatin regulation. Here, we describe the heterologous expression and biochemical characterization of one of these Arabidopsis proteins (AtLSD1) and show that, similarly to HsLSD1, it has demethylase activity toward mono- and dimethylated Lys4 but not dimethylated Lys9 and Lys27 of histone 3. Modeling of the AtLSD1 three-dimensional structure using the HsLSD1 crystal structure as a template revealed a high degree of conservation of the residues building up the active site and some important differences. Among these differences, the most prominent is the lack of the HsLSD1 Tower domain, which has been shown to interact with CoREST and to be indispensable for HsLSD1 demethylase activity. This observation, together with AtLSD1 peculiar surface electrostatic potential distribution, suggests that the molecular partners of AtLSD1 are probably different from those of the human orthologue. PMID- 18393446 TI - Enzyme kinetics and binding studies on inhibitors of MEK protein kinase. AB - Inhibition of the protein kinase, MEK1, is a potential approach for the treatment of cancer. Inhibitors may act by prevention of activation (PoA), which involves interfering with phosphorylation of nonactivated MEK1 by the upstream kinase, B RAF. Modulation also may occur by inhibition of catalysis (IoC) during phosphorylation of the downstream substrate, ERK2, by activated MEK1. Here, five MEK inhibitors are characterized in terms of binding affinity, PoA, and IoC. The compounds are a butadiene (U-0126), an N-alkoxy amide (CI-1040), two CI-1040 analogues (an anthranilic acid and an N-alkyl amide), and a cyanoquinoline. Some compounds give different mechanisms of inhibition (ATP-competitive, noncompetitive, or uncompetitive) in PoA compared to IoC or show a change in potency between the assays. The inhibitors also exhibit different shifts in potency when either PoA or IoC is compared with binding to nonactivated MEK. The inhibitor potency ranking, therefore, is dependent upon the assay format. When the ATP concentration equals K m, IoC IC 50 increases in the order CI-1040 approximately cyanoquinoline < anthranilic acid approximately U-0126 < alkyl amide. Conversely, the K d from nonactivated MEK1 for four of the compounds varies between more than 6-fold lower and over 18-fold higher than this IC 50, with U-0126 having the lowest K d and CI-1040 having the highest. In PoA when the ATP concentration equals K m, U-0126 has the lowest IC 50, becoming more potent than CI-1040, the cyanoquinoline, and the anthranilic acid. These observations have implications for understanding structure-activity relationships of MEK inhibitors and illustrate how assays can be designed to favor different compounds. PMID- 18393447 TI - Monitoring aromatic picosecond to nanosecond dynamics in proteins via 13C relaxation: expanding perturbation mapping of the rigidifying core mutation, V54A, in eglin c. AB - Long-range effects, such as allostery, have evolved in proteins as a means of regulating function via communication between distal sites. An NMR-based perturbation mapping approach was used to more completely probe the dynamic response of the core mutation V54A in the protein eglin c by monitoring changes in picosecond to nanosecond aromatic side-chain dynamics and H/D exchange stabilities. Previous side-chain dynamics studies on this mutant were limited to methyl-bearing residues, most of which were found to rigidify on the picosecond to nanosecond time scale in the form of a contiguous "network". Here, high precision (13)C relaxation data from 13 aromatic side chains were acquired by applying canonical relaxation experiments to a newly developed carbon labeling scheme [Teilum et al. (2006) J. Am. Chem. Soc. 128, 2506-2507]. The fitting of model-free parameters yielded S (2) variability which is intermediate with respect to backbone and methyl-bearing side-chain variability and tau e values that are approximately 1 ns. Inclusion of the aromatic dynamic response results in an expanded network of dynamically coupled residues, with some aromatics showing increases in flexibility, which partially offsets the rigidification in methyl side chains. Using amide hydrogen exchange, dynamic propagation on a slower time scale was probed in response to the V54A perturbation. Surprisingly, regional stabilization (slowed exchange) 10-12 A from the site of mutation was observed despite a global destabilization of 1.5 kcal x mol (-1). Furthermore, this unlikely pocket of stabilized residues colocalizes with increases in aromatic flexibility on the faster time scale. Because the converse is also true (destabilized residues colocalize with rigidification on the fast time scale), a plausible entropy-driven mechanism is discussed for relating colocalization of opposing dynamic trends on vastly different time scales. PMID- 18393448 TI - Charge transfer in the K proton pathway linked to electron transfer to the catalytic site in cytochrome c oxidase. AB - Cytochrome c oxidase couples electron transfer from cytochrome c to O 2 to proton pumping across the membrane. In the initial part of the reaction of the reduced cytochrome c oxidase with O 2, an electron is transferred from heme a to the catalytic site, parallel to the membrane surface. Even though this electron transfer is not linked to proton uptake from solution, recently Belevich et al. [(2006) Nature 440, 829] showed that it is linked to transfer of charge perpendicular to the membrane surface (electrogenic reaction). This electrogenic reaction was attributed to internal transfer of a proton from Glu286, in the D proton pathway, to an unidentified protonatable site "above" the heme groups. The proton transfer was proposed to initiate the sequence of events leading to proton pumping. In this study, we have investigated electrogenic reactions in structural variants of cytochrome c oxidase in which residues in the second, K proton pathway of cytochrome c oxidase were modified. The results indicate that the electrogenic reaction linked to electron transfer to the catalytic site originates from charge transfer within the K pathway, which presumably facilitates reduction of the site. PMID- 18393450 TI - Multidomain initiation factor 2 from Thermus thermophilus consists of the individual autonomous domains. AB - The three-dimensional chalice-like crystal structure of initiation factor 2 IF2/eIF5B from Methanobacterium thermoautotrophicum represents a novel fold and domain architecture in which the N-terminal G domain and the C-terminal C domain are separated by an approximately 40 A alpha-helix. Homologous Thermus thermophilus initiation factor 2 (IF2wt), G (IF2G), and C (IF2C) domains were successfully overexpressed and purified which enabled us to perform a thermodynamic analysis and to asses the role of the domain architecture in this atypical fold. Circular dichroism in the far-UV region demonstrated that the proteins are well-folded and that the secondary structure content resembles that of IF2 from M. thermoautotrophicum. IF2wt and IF2G are monomeric proteins, while IF2C has a tendency to form dimeric species as shown by sedimentation velocity studies on analytical ultracentrifugation and differential scanning calorimetry scan analysis. Thermal denaturation studies of multidomain IF2wt reveals an exceptionally high reversibility (>90%) of the transition with a melting temperature of 94.5 degrees C. Melting temperature of IF2wt may be further increased in the presence of its physiological ligand GDP and the GTP analogue, GppNHp. The high reversibility of denaturation is achieved by the modular structure of the protein and by the high reversibility of the thermal denaturation of IF2G. On the other hand, hydrophobic IF2C aggregates during the thermal transition, and the aggregation is suppressed by guanidine hydrochloride. Isothermal denaturation demonstrates that both IF2G and IF2C have comparable stabilities of 46 and 33 kJ/mol, respectively. The apparent cooperative unfolding of the full-length protein has an unusually small denaturant m value. This together with the phase diagram method of analysis indicates the presence of intermediate(s) due to the independent unfolding of IF2G and IF2C. Despite an absence of apparent interactions between the domains in vitro, IF2G plays a role in IF2C reversibility in thermal denaturation. In conclusion, interactions between the domains of folded IF2wt in vivo are likely mediated by their alpha helix connection and/or by a conformational change on the ribosome. PMID- 18393449 TI - Human quiescin-sulfhydryl oxidase, QSOX1: probing internal redox steps by mutagenesis. AB - The flavoprotein quiescin-sulfhydryl oxidase (QSOX) rapidly inserts disulfide bonds into unfolded, reduced proteins with the concomitant reduction of oxygen to hydrogen peroxide. This study reports the first heterologous expression and enzymological characterization of a human QSOX1 isoform. Like QSOX isolated from avian egg white, recombinant HsQSOX1 is highly active toward reduced ribonuclease A (RNase) and dithiothreitol but shows a >100-fold lower k cat/ K m for reduced glutathione. Previous studies on avian QSOX led to a model in which reducing equivalents were proposed to relay through the enzyme from the first thioredoxin domain (C70-C73) to a distal disulfide (C509-C512), then across the dimer interface to the FAD-proximal disulfide (C449-C452), and finally to the FAD. The present work shows that, unlike the native avian enzyme, HsQSOX1 is monomeric. The recombinant expression system enabled construction of the first cysteine mutants for mechanistic dissection of this enzyme family. Activity assays with mutant HsQSOX1 indicated that the conserved distal C509-C512 disulfide is dispensable for the oxidation of reduced RNase or dithiothreitol. The four other cysteine residues chosen for mutagenesis, C70, C73, C449, and C452, are all crucial for efficient oxidation of reduced RNase. C452, of the proximal disulfide, is shown to be the charge-transfer donor to the flavin ring of QSOX, and its partner, C449, is expected to be the interchange thiol, forming a mixed disulfide with C70 in the thioredoxin domain. These data demonstrate that all the internal redox steps occur within the same polypeptide chain of mammalian QSOX and commence with a direct interaction between the reduced thioredoxin domain and the proximal disulfide of the Erv/ALR domain. PMID- 18393451 TI - Accelerated cytotoxic mechanism screening of hydralazine using an in vitro hepatocyte inflammatory cell peroxidase model. AB - Long-term treatment of hypertensive disorders with hydralazine has resulted in some patients developing hepatitis and lupus erythematosus, an autoimmune syndrome. The concentration of hydralazine required to cause 50% cytotoxicity in 2 h (LC(50)) toward isolated rat hepatocytes was found to be 8 mM. Cytotoxicity was delayed by the P450 inhibitor, 1-aminobenzotriazole, suggesting that P450 catalyzed the formation of toxic metabolites from hydralazine. No hydralazine induced oxidative stress was apparent as there was little effect on hepatocyte lipid peroxidation, protein carbonyl formation, intracellular H(2)O(2), or hepatocyte GSH levels and no effect of butylated hydroxyanisole (BHA) on cytotoxicity. Drug-induced hepatotoxicity in vivo has often been attributed to infiltrating inflammatory cells, for example, neutrophils or resident Kupffer cells whose NADPH oxidase generates H(2)O(2), when activated. The effect of a nontoxic continuous infusion of H(2)O(2) on hydralazine cytotoxicity was investigated. It was found that H(2)O(2) increased hepatocyte susceptibility to hydralazine 4-fold (LC(50), 2 mM). Cytotoxicity was still prevented by the P450 inhibitor but now involved some oxidative stress as shown by increased protein carbonyls, endogenous H(2)O(2), and GSH oxidation. Lipid peroxidation was not increased, and cytotoxicity was not inhibited by BHA. Cytotoxicity, however, was inhibited by 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), a ROS scavenger. Because neutrophils or Kupffer cells release myeloperoxidase on activation, the effect of adding peroxidase to the hepatocytes exposed to H(2)O(2) on hydralazine cytotoxicity was investigated. It was found that peroxidase/H(2)O(2) increased hepatocyte susceptibility to hydralazine 80-fold (LC 50, 0.1 mM). Furthermore, cytotoxicity occurred following extensive oxidative stress that included lipid peroxidation, and cytotoxicity that was now prevented by the antioxidant BHA. These results indicate that three cytotoxic pathways exist for hydralazine: a P450-catalyzed pathway not involving oxidative stress, a P450/H(2)O(2)-catalyzed oxidative stress-mediated cytotoxic pathway not involving lipid peroxidation, and a peroxidase/H(2)O(2)-catalyzed lipid peroxidation mediated cytotoxic pathway. PMID- 18393453 TI - Bioactivation of a dihydropyrazole-1-carboxylic acid-(4-chlorophenyl amide) scaffold to a putative p-chlorophenyl isocyanate in rat liver microsomes and in vivo in rats. AB - Compound I (4,5-dihydropyrazole-1,5-dicarboxylic acid-1-[(4-chlorophenyl)-amide] 5-[(2-oxo-2 H-[1,3']bipyridinyl-6'-yl)-amide] was found to undergo metabolic activation in rat liver microsomes in the presence of NADPH. A reactive intermediate, postulated to be p-chlorophenyl isocyanate (CPIC), was trapped by GSH in vitro and characterized by liquid chromatography tandem mass spectrometry (LC/MS/MS). Subsequently, the structure of the GSH conjugate was confirmed by a comparison with a synthetic standard. The GSH conjugate was also found in the bile of rats that received an oral dose (10 mg/kg) of compound I. Further analyses of rat bile and urine using online electrochemical derivatization coupled to LC/MS demonstrated the presence of p-chlorophenyl aniline (CPA), a hydrolytic product of the intermediate isocyanate. This provided further evidence for the potential existence of CPIC. Approximately 7% of the dose was accounted by the products of CPIC, which included the GSH conjugate and CPA excreted in bile and urine. Multiple rat cytochrome P450 enzymes, including P450 1A, P450 2C, and P450 3A, appeared to be responsible for the activation of compound I to CPIC. The activation kinetics of compound I to CPIC in male rat liver microsomes exhibited a biphasic profile, indicative of at least two contributing P450 enzymes. One enzyme showed a small value of K m at 42 microM and a low V max of 66 pmol min (-1) mg (-1), while the other exhibited a large value of K m at 148 microM and a high V max of 1200 pmol min (-1) mg (-1). The formation of a putative CPIC intermediate, a carbamoylating species known to be capable of covalent binding to macromolecules, suggests a potential liability associated with the compound, particularly the dihydropyrazole-1-carboxylic acid-(4 chlorophenyl amide) scaffold, which appears to be responsible for the generation of CPIC. The mechanism of bioactivation to the putative CPIC is postulated to involve an initial P450-mediated hydroxylation of the pyrazoline at the 3 position followed by subsequent decomposition to CPIC. This mechanistic insight into the bioactivation allowed for the development of a rational structural modification strategy to mitigate or minimize the reactive metabolite formation. One of the approaches included the introduction of a metabolically stable substituent with electron-donating character at the 3 position of pyrazoline to block CPIC formation. PMID- 18393452 TI - Mitochondrial DNA impairment in nucleoside reverse transcriptase inhibitor associated cardiomyopathy. AB - Acquired immune deficiency syndrome (AIDS) is a global epidemic that continues to escalate. Recent World Health Organization estimates include over 33 million people currently diagnosed with HIV/AIDS. Another 20 million HIV-infected individuals died over the past quarter century. Antiretrovirals are effective treatments that changed the outcome of HIV infection from a fatal disease to a chronic illness. Cardiomyopathy (CM) is a bona fide component of HIV/AIDS with occurrence that is higher in HIV positive individuals. CM may result from individual or combined effects of HIV, immune reactions, or toxicities of prolonged antiretrovirals. Nucleoside reverse transcriptase inhibitors (NRTIs) are the cornerstone of antiretroviral therapy. Despite pharmacological benefits of NRTIs, NRTI side effects include increased risk for CM. Clinical observations and in vitro and in vivo studies support various mechanisms of CM. This perspective highlights some of the hypotheses and focuses on mitochondrial associated pathways of NRTI- related CM. PMID- 18393454 TI - High-throughput method for ranking the affinity of peptide ligands selected from phage display libraries. AB - The use of phage display peptide libraries allows rapid isolation of peptide ligands for any target selector molecule. However, due to differences in peptide expression and the heterogeneity of the phage preparations, there is no easy way to compare the binding properties of the selected clones, which operates as a major "bottleneck" of the technology. Here, we present the development of a new type of library that allows rapid comparison of the relative affinity of the selected peptides in a high-throughput screening format. As a model system, a phage display peptide library constructed on a phagemid vector that contains the bacterial alkaline phosphatase gene (BAP) was selected with an antiherbicide antibody. Due to the intrinsic switching capacity of the library, the selected peptides were transferred "en masse" from the phage coat protein to BAP. This was coupled to an optimized affinity ELISA where normalized amounts of the peptide BAP fusion allow direct comparison of the binding properties of hundreds of peptide ligands. The system was validated by plasmon surface resonance experiments using synthetic peptides, showing that the method discriminates among the affinities of the peptides within 3 orders of magnitude. In addition, the peptide-BAP protein can find direct application as a tracer reagent. PMID- 18393456 TI - A concentration-dependent analysis method for high density protein microarrays. AB - Protein microarray technology is rapidly growing and has the potential to accelerate the discovery of targets of serum antibody responses in cancer, autoimmunity and infectious disease. Analytical tools for interpreting this high throughput array data, however, are not well-established. We developed a concentration-dependent analysis (CDA) method which normalizes protein microarray data based on the concentration of spotted probes. We show that this analysis samples a data space that is complementary to other commonly employed analyses, and demonstrate experimental validation of 92% of hits identified by the intersection of CDA with other tools. These data support the use of CDA either as a preprocessing step for a more complete proteomic microarray data analysis or as a stand-alone analysis method. PMID- 18393455 TI - Factors determining the efficacy of nuclear delivery of antisense oligonucleotides by gold nanoparticles. AB - The present study investigates the applicability of nanoparticle delivery vectors for two-stage targeting that involves both cell entry by endocytosis and nuclear targeting using viral peptide signals. A nanoparticle vector consists of four components: a carrier nanoparticle, a stabilizer, targeting peptides, and a therapeutic cargo. Extensive study of bovine serum albumin (BSA)-peptide stabilized nanoparticle conjugates demonstrated limitations of these systems due to colloidal instability when oligonucleotides and multiple peptides were attached to the BSA protein. We found that the widely used protein streptavidin (SA) was an appropriate alternative to BSA for cell-targeting experiments. Targeting peptides and gene splicing oligonucleotides were attached to SA nanoparticles using biotin labels. The present study uses a gene-splicing assay as a test for oligonucleotide delivery to the cell nucleus. Successful modification of gene splicing by an antisense oligonucleotide indicates that the latter must have crossed the plasma membrane, entered the nucleus, found the target sequence in the newly transcribed pre-mRNA, and hybridized to it in the spliceosome strongly enough to displace the splicing factors designed to interact with the target sequence. Targeting nanoparticles that carry gene-splicing oligonucleotides were compared with a control experiment that used lipofectamine (LF). While enhanced activity was observed in the control experiment, in the presence of LF, no gene splicing was observed for the nanoparticle targeting vectors without LF. We conclude that sequestration of cargo from the harsh conditions of the endosome is a desirable strategy for cell-targeting nanoparticles. PMID- 18393457 TI - Proteomics as a complementary tool for identifying unintended side effects occurring in transgenic maize seeds as a result of genetic modifications. AB - To improve the probability of detecting unintended side effects during maize gene manipulations by bombardment, proteomics was used as an analytical tool complementary to the existing safety assessment techniques. Since seed proteome is highly dynamic, depending on the species variability and environmental influence, we analyzed the proteomic profiles of one transgenic maize variety (event MON 810) in two subsequent generations (T05 and T06) with their respective isogenic controls (WT05 and WT06). Thus, by comparing the proteomic profiles of WT05 with WT06 we could determine the environmental effects, while the comparison between WT06 and T06 seeds from plants grown under controlled conditions enabled us to investigate the effects of DNA manipulation. Finally, by comparison of T05 with T06 seed proteomes, it was possible to get some indications about similarities and differences between the adaptations of transgenic and isogenic plants to the same strictly controlled growth environment. Approximately 100 total proteins resulted differentially modulated in the expression level as a consequence of the environmental influence (WT06 vs WT05), whereas 43 proteins resulted up- or down-regulated in transgenic seeds with respect to their controls (T06 vs WT06), which could be specifically related to the insertion of a single gene into a maize genome by particle bombardment. Transgenic seeds responded differentially to the same environment as compared to their respective isogenic controls, as a result of the genome rearrangement derived from gene insertion. To conclude, an exhaustive differential proteomic analysis allows to determine similarities and differences between traditional food and new products (substantial equivalence), and a case-by-case assessment of the new food should be carried out in order to have a wide knowledge of its features. PMID- 18393458 TI - Aqueous polysaccharide associations mediated by beta-cyclodextrin polymers. AB - Macromolecular assemblies were elaborated by mixing in water hydrophobically modified dextrans (MDC(n)) and beta-cyclodextrin polymers (pbetaCD) interacting by inclusion complexation between the hydrophobic moieties of MDCn and the beta cyclodextrin cavities of pbetaCD. Dextrans have been modified by grafting alkyl groups (C(n)) of varying chain lengths (n = 8-16) and grafting ratio (3-6 mol%). Different pbetaCD polymers were synthesized by polycondensation of beta cyclodextrin and epichlorohydrin. The polymer-polymer interactions have been studied by fluorimetry, isothermal titration microcalorimetry, phase diagrams, and viscosimetry. The viscoelastic properties of the temporary networks (in the semidilute range) have been studied by rheology. The interaction mechanisms between the MDCn and pbetaCD can be understood taking into account the strength of the interaction between the alkyl group and the beta-cyclodextrin cavity (mainly controlled by the alkyl chain length), the density of junctions between the chains (depending on the alkyl grafting density and the pbetaCD molecular weight), and additional cooperative effect (arising for high alkyl grafting density). PMID- 18393459 TI - A novel one-pot pseudo-five-component synthesis of 4,5,6,7-tetrahydro-1H-1,4 diazepine-5-carboxamide derivatives. AB - A novel one-pot pseudo-five-component synthesis of 4,5,6,7-tetrahydro-1 H-1,4 diazepine-5-carboxamide derivatives starting from simple and readily available inputs including 2,3-diaminomaleonitrile, a cyclic or acyclic ketone, an isocyanide, and water in the presence of a catalytic amount of p-toluenesulfonic acid in aqueous medium at ambient temperature in high yields is described. PMID- 18393460 TI - Ionic liquid, surrogate hydrogen bromide reagent for ring opening of cyclopropyl ketones. AB - Ionic liquid reagents created by addition of 1 equiv of either CF3CO2H or CF3SO3H to N-pentylpyridinium bromide exhibit excellent chemical reactivities as surrogate HBr reagents in ring-opening reactions of cyclopropyl ketones as well as of 2,2-difluorocyclopropyl ketones to form the respective 3-bromopropyl or 3 bromo-2,2-difluoropropyl ketones in good to excellent yields. PMID- 18393461 TI - Sesquiterpene lactones from Staehelina fruticosa. AB - The phytochemical analysis of Staehelina fruticosa led to the isolation of four germacranolide-type sesquiterpene lactones (1- 4), including two new glycosides. The structures of these sesquiterpene lactones were elucidated using spectroscopic techniques, and enzymatic hydrolysis was carried out to confirm the nature of the two glycoside derivatives. Molecular modeling was incorporated to substantiate their relative configuration. PMID- 18393462 TI - Determination of analyte concentration using the residual solvent resonance in (1)H NMR spectroscopy. AB - An NMR protocol that uses the residual proton signal from DMSO -d(6) (i.e., DMSO d(5)) to determine the concentration of an analyte in a NMR sample was developed. This technique provides an alternative method for determining the molar concentration of compounds in solution without prior knowledge of their molecular weight. The method is particularly useful when submilligram quantities of compound are to be analyzed and is applicable to a variety of different research areas such as compound management, and natural product, combinatorial, and medicinal chemistry. PMID- 18393463 TI - Parnapimarol and nepetaparnone from Nepeta parnassica. AB - Parnapimarol (1), a new pimarane diterpene, along with nepetaparnone (2) and nepetanudone (3), one new and one previously reported nepetalactone dimer, respectively, were isolated from the dichloromethane extract of the aerial parts of Nepeta parnassica, collected on Mt. Parnassos, Greece. The structures and relative configurations of 1-3 were determined on the basis of their spectroscopic characteristics (1D and 2D NMR, IR, MS). The structure of 2 was confirmed by single-crystal X-ray diffraction analysis. The insecticidal activity of 1-3 against ants and mosquito larvae was also evaluated. PMID- 18393464 TI - Aplysamine 6, an alkaloidal inhibitor of Isoprenylcysteine carboxyl methyltransferase from the sponge Pseudoceratina sp. AB - The anticancer target isoprenylcysteine carboxyl methyltransferase (Icmt) was the focus of a natural product high-throughput screening campaign. The Australian marine sponge Pseudoceratina sp. yielded aplysamine 6, a new bromotyrosine derivative with an alpha,beta-unsaturated amide linkage, as the bioactive constituent. Its structure was determined by 1D and 2D NMR spectroscopy. PMID- 18393465 TI - Dragonamides C and D, linear lipopeptides from the marine cyanobacterium brown Lyngbya polychroa. AB - Two new linear lipopeptides, 1 and 2, and a known compound, curacin D, have been isolated from a marine cyanobacterium, brown Lyngbya polychroa, collected from Hollywood Beach, Fort Lauderdale, Florida. Their planar structures were elucidated by 1D and 2D NMR techniques, and absolute configurations were assigned using chiral HPLC. The new compounds were assigned the trivial names dragonamide C (1) and dragonamide D (2), as their peptide moiety is related to previously reported dragonamides A and B. PMID- 18393466 TI - Sodium-coupled neurotransmitter transporters. PMID- 18393467 TI - Microwave-assisted efficient copper-promoted N-arylation of amines with arylboronic acids. PMID- 18393468 TI - Field-dependent DNA mobility in 20 nm high nanoslits. AB - The transport behavior of lambda-DNA (48 kbp) in fused silica nanoslits is investigated upon application of electrical fields of different strengths. The slit dimensions are 20 nm in height, 3 microm in width, and 500 microm in length. With fields of 30 kV/m or below, the molecules move fluently through the slits, while at higher electrical fields, the DNA molecules move intermittently, resulting in a strongly reduced mobility. We propose that the behavior can be explained by mechanical and/or field-induced dielectrophoretic DNA trapping due to the surface roughness in the nanoslits. The observation of preferential pathways and trapping sites of the lambda-DNA molecules through the nanoslits supports this hypothesis and indicates that the classical viscous friction models to explain the DNA movement in nanoslits needs to be modified to include these effects. Preliminary experiments with the smaller XbaI-digested litmus-DNA (2.8 kbp) show that the behavior is size-dependent, suggesting that the high field electrophoresis in nanoslits can be used for DNA separation. PMID- 18393470 TI - Three-dimensionally-patterned submicrometer-scale hydrogel/air networks that offer a new platform for biomedical applications. AB - Phase mask interference lithography was employed to fabricate three-dimensional (3D) hydrogel structures with high surface area on neural prosthetic devices. A random terpolymer of poly(hydroxyethyl methacrylate- ran-methyl methacrylate- ran methacrylic acid) was synthesized and used as a negative-tone photoresist to generate bicontinuous 3D hydrogel structures at the submicrometer scale. We demonstrated that the fully open 3D hydrogel/air networks can be used as a pH responsive polymeric drug-release system for the delivery of neurotrophins to enhance the performance of neural prosthetic devices. Additionally an open hydrogel structure will provide direct access of neuronal growth to the device for improved electrical coupling. PMID- 18393469 TI - UV-light induced fabrication of CdCl2 nanotubes through CdSe/Te nanocrystals based on dimension and configuration control. AB - Since the discovery of WS2 nanotubes in 1992 ( Nature 1992, 360, 444), there have been significant research efforts to synthesize nanotubes and fullerene-like hollow nanoparticles (HNPs) of inorganic materials ( Nat. Nanotechnol. 2006, 1, 103) due to their potential applications as solid lubrications ( J. Mater. Chem. 2005, 15, 1782), chemical sensing ( Adv. Funct. Mater. 2006, 16, 371), drug delivering ( J. Am. Chem. Soc. 2005, 127, 7316), catalysis ( Adv. Mater. 2006, 18, 2561), or quantum harvesting ( Acc. Chem. Res. 2006, 39, 239). Nanotubes can be produced either by rolling up directly from layer compounds ( Nature 2001, 410, 168) or through other mechanisms ( Adv. Mater. 2004, 16, 1497) such as template growth ( Nature 2003, 422, 599) and decomposition ( J. Am. Chem. Soc. 2001, 123, 4841). The Kirkendall effect, a classical phenomenon in metallurgy ( Trans. AIME 1947, 171, 130), was recently exploited to fabricate hollow 0-D nanocrystals ( Science 2004, 304, 711) as well as 1-D nanotubes ( Nat. Mater. 2006, 5, 627). Although the dimension of resulting hollow nanostructures depends on precursors, the hollow nanomaterials can also be organized into various dimensional nanostructures spontaneously or induced by an external field. In this letter, we report, for the first time, the UV-light induced fabrication of the ends-closed 1-D CdCl2 nanotubes from 0-D CdSe solid nanocrystals through the Kirkendall effect and the head-to-end assembled process. Our results demonstrate the possibility to control the dimension (0-D to 1-D) and the configuration (solid to hollow) of nanostructures simultaneously and have implications in fabricating hollow nano-objects from zero-dimensional to multidimensional. PMID- 18393471 TI - Localized surface plasmon resonances in aluminum nanodisks. AB - The plasmonic properties of arrays of supported Al nanodisks, fabricated by hole mask colloidal lithography (HCL), are analyzed for the disk diameter range 61-492 nm at a constant disk height of 20 nm. Strong and well-defined (UV-vis-NIR) localized surface plasmon resonances are found and experimentally characterized with respect to spectral peak positions, peak widths, total cross sections, and radiative and nonradiative decay channels. Theoretically, the plasmon excitations are described by electrostatic spheroid theory. Very good qualitative and quantitative agreement between model and experiment is found for all these observables by assuming a nanoparticle embedded in a few nanometer thick homogeneous (native) aluminum oxide shell. Other addressed aspects are: (i) the role of the strong interband transition in Al metal, located at 1.5 eV, for the plasmonic excitations of Al nanoparticles, (ii) the role of the native oxide layer, and (iii) the possibility of using the plasmon excitation as an ultrasensitive, remote, real-time probe for studies of oxidation/corrosion kinetics in metal nanoparticle systems. PMID- 18393472 TI - Photocurrent polarization anisotropy of randomly oriented nanowire networks. AB - While the polarization sensitivity of single or aligned NW ensembles is well known, this article reports on the existence of residual photocurrent polarization sensitivities in random NW networks. In these studies, CdSe and CdTe NWs were deposited onto glass substrates and contacted with Au electrodes separated by 30-110 microm gaps. SEM and AFM images of resulting devices show isotropically distributed NWs between the electrodes. Complementary high resolution TEM micrographs reveal component NWs to be highly crystalline with diameters between 10 and 20 nm and with lengths ranging from 1 to 10 microm. When illuminated with visible (linearly polarized) light, such random NW networks exhibit significant photocurrent anisotropies rho = 0.25 (sigma = 0.04) [rho = 0.22 (sigma = 0.04)] for CdSe (CdTe) NWs. Corresponding bandwidth measurements yield device polarization sensitivities up to 100 Hz. Additional studies have investigated the effects of varying the electrode potential, gap width, and spatial excitation profile. These experiments suggest electrode orientation as the determining factor behind the polarization sensitivity of NW devices. A simple geometric model has been developed to qualitatively explain the phenomenon. The main conclusion from these studies, however, is that polarization sensitive devices can be made from random NW networks without the need to align component wires. PMID- 18393473 TI - Optical biosensor based on hollow integrated waveguides. AB - The first absorbance biosensor based on pure silicon hollow integrated waveguides is presented in this work. With the use of horseradish peroxidase (HRP) as a model recognition element, an enzymatic sensor for the measurement of hydrogen peroxide was fabricated, numerically simulated, and experimentally characterized. Waveguides with widths ranging from 50 to 80 microm, having a depth of 50 microm and lengths up to 5 mm were easily fabricated by just one photolithographic step. These were further modified by covalent immobilization of HRP using silanization chemistry. Simulation studies of the proposed approach showed a sensor linear behavior up to 300 microM H2O2 and a sensitivity of 2.7 x 10(-3) AU/microM. Experimental results were in good agreement with the simulated ones. A linear behavior between 10 and 300 microM H2O2, a sensitivity of 3 x 10(-3) AU/microM, and a signal-to-noise ratio around 20 dB were attained. Also, kinetic studies of the activity of the immobilized enzyme on the silicon waveguide surface gave an apparent Michaelis-Menten constant of 0.44 mM. The simple technology proposed in this work enables the fabrication of cost-effective, easy-to-use, miniaturized biosensor generic platforms, these being envisioned as excellent candidates for the development of lab-on-a-chip systems. PMID- 18393474 TI - Assessment of the accuracy of density functionals for prediction of relative energies and geometries of low-lying isomers of water hexamers. AB - Water hexamers provide a critical testing ground for validating potential energy surface predictions because they contain structural motifs not present in smaller clusters. We tested the ability of 11 density functionals (four of which are local and seven of which are nonlocal) to accurately predict the relative energies of a series of low-lying water hexamers, relative to the CCSD(T)/aug'-cc pVTZ level of theory, where CCSD(T) denotes coupled cluster theory with an interative treatment of single and double excitations and a quasi-perturbative treatment of connected triple excitations. Five of the density functionals were tested with two different basis sets, making a total of 16 levels of density functional theory (DFT) tested. When single-point energy calculations are carried out on geometries obtained with second-order Moller-Plesset perturbation theory (MP2), only three density functionals, M06-L, M05-2X, and M06-2X, are able to correctly predict the relative energy ordering of the hexamers. These three functionals predict that the range of energies spanned by the six isomers is 3.2 5.6 kcal/mol, whereas the other eight functionals predict ranges of 1.0-2.4 kcal/mol; the benchmark value for this range is 3.1 kcal/mol. When the hexamers are optimized at each level of theory, all methods are able to reproduce the MP2 geometries well for all isomers except the boat and bag isomers, and DFT optimization changes the energy ordering for seven of the 16 methods tested. The addition of zero-point energy changes the energy ordering for all of the density functionals studied except for M05-2X and M06-2X. The variation in relative energies predicted by the different methods highlights the necessity for exercising caution in the choice of density functionals used in future studies. Of the 11 density functionals tested, the most accurate results for energies were obtained with the PWB6K, MPWB1K, and M05-2X functionals. PMID- 18393475 TI - Ground- and excited-state infrared spectra of an azacrown-substituted [(bpy)Re(CO)3L]+ complex: structure and bonding in ground and excited states and effects of Ba2+ binding. AB - Ground- and excited-state infrared spectra are reported for a [(bpy)ReI(CO)3L]+ complex (bpy = 2,2'-bipyridine) in which L contains an azacrown ether that is linked to Re via an amidopyridyl group. Ground-state band assignments are made with the aid of spectra from model complexes in which a similar electron-donating dimethylamino group replaces the azacrown, in which an electron-donor group is absent, and from the L ligands, in conjunction with DFT calculations. Picosecond time-resolved IR (TRIR) spectra in the nu(CO) region show bands characteristic of a metal-to-ligand charge-transfer (MLCT) excited state, [(bpy*-)ReII(CO)3L]+, from the complex in which an electron-donor group is absent, whereas those from the azacrown complex show bands of an MLCT state evolving into those characteristic of a ligand-to-ligand charge-transfer (LLCT) excited state, [(bpy* )ReI(CO)3(L*+)]+, formed upon intramolecular electron transfer. Picosecond TRIR spectra of the azacrown complex in the fingerprint region show strong L ligand bands that indicate that significant charge redistribution occurs within this ligand in the MLCT state and that decay as the LLCT state forms. Picosecond TRIR spectra obtained when Ba2+ was complexed to the azacrown show bands of only an MLCT state at all times up to 2 ns, consistent with the presence of Ba2+ inhibiting electron transfer from the azacrown N atom to form the LLCT state, and the positions of the bands in the fingerprint region provide direct evidence for the proposal that charge redistribution within the L ligand induces Ba2+ release from the azacrown in the MLCT state. PMID- 18393476 TI - Theoretical investigation of the magnetic interactions of Ni9 complexes. AB - On the basis of density-functional theory (DFT) calculations, a theoretical analysis of the exchange interactions in Ni9L2(O2CMe)8{(2-py)2CO2}4, was performed, where L is a bridging ligand, OH- (1) or N3- (2). Each magnetic interaction between the Ni spin centers is analyzed for 1 and 2 in terms of exchange integrals (J values), orbital overlap integrals (T values) and natural orbitals. It was found that a J3 interaction, which is a magnetic interaction via the bridging ligand orbitals, mainly controls the whole magnetic properties, and the dominant interaction is a sigma-type orbital interaction between Ni dz2 orbitals. Further investigations on the magnetostructural correlations are performed on the J3 interactions using simplest Ni-L-Ni models. These models reproduced the magnetic interactions qualitatively well not only for the Ni9 complexes but also for other inorganic complexes. Strong correlations have been found between the magnetic orbital overlaps (T values) and the Ni-L-Ni angle. These results revealed that the difference of the magnetic properties between OH- and N3- is caused by the orbital overlap integral (T values) of the sigma-type J3 interaction pathway. The magnetic interactions are also discussed from a Hubbard model by evaluating the transfer integral (t) and on-site Coulomb integrals (U), in relation to the Heisenberg picture. PMID- 18393477 TI - Photoinduced triplet-triplet energy transfer via the 2-ureido-4[1H]-pyrimidinone self-complementary quadruple hydrogen-bonded module. AB - Phosphorescence quenching and flash photolysis experiments demonstrate that photoinduced intra-assembly triplet-triplet energy transfer can take place via a 2-ureido-4[1H]-pyrimidinone-bridged benzophenone-naphthalene assembly I with a rate constant of 3.0 x 106 s-1 and an efficiency of 95% in CH2Cl2. This new finding suggests that with high binding strength and directionality, the 2-ureido 4[1H]-pyrimidinone hydrogen-bonded module may serve as a new model to illustrate the fundamental principles governing the triplet-triplet energy-transfer process through hydrogen bonds. PMID- 18393478 TI - Theoretical study of the decomposition reactions in substituted nitrobenzenes. AB - The influence of substituent nature and position on the unimolecular decomposition of nitroaromatic compounds was investigated using the density functional theory at a PBE0/6-31+G(d,p) level. As the starting point, the two main reaction paths for the decomposition of nitrobenzene were analyzed: the direct carbon nitrogen dissociation (C6H5 + NO2) and a two step mechanism leading to the formation of phenoxyl and nitro radicals (C6H5O + NO). The dissociation energy of the former reaction was calculated to be 7.5 kcal/mol lower than the activation energy of the second reaction. Then the Gibbs free energies were computed for 15 nitrobenzene derivatives characterized by different substituents (nitro, methyl, amino, carboxylic acid, and hydroxyl) in the ortho, meta, and para positions. In meta position, no significant changes appeared in the reaction energy profiles whereas ortho and para substitutions led to significant deviations in energies on the decomposition mechanisms due to the resonance effect of the nitro group without changing the competition between these mechanisms. In the case of para and meta substitutions, the carbon-nitro bond dissociation energy has been directly related to the Hammett constant as an indicator of the electron donor-acceptor effect of substituents. PMID- 18393479 TI - High levels of electrochemical doping of carbon nanotubes: evidence for a transition from double-layer charging to intercalation and functionalization. AB - We studied the transition from the electrochemical double-layer charging regime to intercalative doping of bundled single-walled carbon nanotubes (SWNT) in KCl and HCl aqueous solution. For this purpose we used high doping levels by applying constant potentials above 1000 mV approaching and slightly exceeding the oxidation potential for Cl(-) ions. At each potential in situ Raman measurements of the radial breathing mode (RBM), the high-energy tangential mode (HEM), and the disorder-induced (D) mode were performed. Furthermore, the conductivity and reflectivity of a set of SWNT samples were measured as a function of doping and subsequently the samples were examined by X-ray photoelectron spectroscopy (XPS). From a comparative analysis of the results we conclude that above 1000 mV a significant penetration of chlorine species into the interstitial channels of the SWNT bundles and possible covalent functionalization take place. PMID- 18393480 TI - Crystallization and melting transitions of hexadecane droplets in polystyrene nanocapsules. AB - Shifts to lower transition temperatures are observed for the freezing and melting of submicron-sized hexadecane droplets encapsulated within thin polystyrene shells. Supercooling of approximately 14 K is observed in the first-time cooling scans for the oil. We attribute this lowering predominantly to nucleation of the phase change originating from the oil/polymer/water interface. We obtain a rough estimate of the interfacial tension between the hexadecane oil droplet and the polystyrene of 14 mN/m assuming the Gibbs-Thomson relationship. Melting points for the hexadecane are 1-2 K below the bulk transition temperatures. This effect is connected with the surface/volume ratio of the capsules. Both the supercooling and the melting point depression approach the bulk-phase transition temperature when the sample is taken through multiple cool/heat cycles. The heating and cooling rates affects the number of cycles required before bulk-like behavior is observed. The thickness of the capsule wall is also observed to be critical to how many cooling cycles are required. Two hypotheses to explain this behavior are presented. PMID- 18393481 TI - Selenourea-Ca2+ reactions in gas phase. Similarities and dissimilarities with urea and thiourea. AB - The gas-phase reactions between Ca(2+) and selenourea were investigated by means of electrospray/tandem mass spectrometry techniques. The MS/MS spectra of [Ca(selenourea)](2+) complexes show intense peaks at m/z 43, 121, 124, and 146 and assigned to monocations produced in different coulomb explosion processes. The structures and bonding characteristics of the stationary points of the [Ca(selenourea)](2+) potential energy surface (PES) were theoretically studied by DFT calculations carried out at the B3LYP/6-311+G(3df,2p)//B3LYP/6-311+G(d,p) level. The analysis of the topology of this PES allows identification of H(2)NCNH(+), CaSeH(+), selenourea(+). and CaNCSe(+) ion peaks at m/z 43, 121, 124, and 146, respectively. The reactivity of selenourea and the topology of the corresponding potential energy surface mimic that of thiourea. However, significant dissimilarities are found with respect to urea. The dissociative electron-transfer processes, not observed for urea, is one of the dominant fragmentations for selenourea, reflecting its much lower ionization energy. Similarly, the coulomb explosions yielding CaXH(+) + H(2)NCNH(+) (X = O or Se), which for urea are not observed, are very favorable for selenourea. Finally, while in urea the loss of NH(3) competes with the formation of NH(4+), for selenourea the latter process is clearly dominant. PMID- 18393482 TI - Reduction of 2,3,5-triphenyl-2H-tetrazolium chloride in the presence of polyelectrolytes containing 4-styrenesulfonate moieties. AB - The redox behavior of 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) in the presence of different polyelectrolytes such as poly(sodium 4-styrenesulfonate) (PSS), poly(sodium 4-styrenesulfonate-co-sodium maleate) at two different comonomer compositions (P(SS(1)-co-MA(1)) and P(SS(3)-co-MA(1))), poly(sodium acrylate-co-sodium maleate) (P(AA(1)-co-MA(1))), and poly(sodium acrylate) (PAA) is studied. Due to aromatic-aromatic interactions, the polyelectrolytes containing benzene sulfonate groups produce a decrease on the reduction rate of TTC in the presence of ascorbic acid (ASC) and a shift of the anodic and cathodic peaks to higher negative potentials for the electrochemical reaction of TTC. As an important conclusion, these effects are a function of the linear aromatic density of the polyelectrolytes. PMID- 18393483 TI - Direct conformational analysis of a 10 nm long oligothiophene wire. AB - Conformational variations of a 10 nm long oligothiophene wire comprising 24 thiophene rings on Au(111), which are related to the various straight and bent shapes of the long wires, have been directly visualized by scanning tunneling microscopy (STM). The local bending angles within the wire are well characterized as s-cis/s-trans configurations of individual thiophene rings. We find that the partial stabilization of the metastable s-cis conformation results in the wire bending, which should be influenced by solvent and substituents. PMID- 18393484 TI - The wettability of fluoropolymer surfaces: influence of surface dipoles. AB - The wettabilities of fluorinated polymers were evaluated using a series of contacting probe liquids ranging in nature from nonpolar aprotic to polar aprotic to polar protic. Fully fluorinated polymers were wet less than partially fluorinated polymers, highlighting the weak dispersive interactions of fluorocarbons. For partially fluorinated polymers, the interactions between the distributed dipoles along the polymer backbone and the dipoles of the contacting liquids were evaluated using both polar and nonpolar probe liquids. The results demonstrate that the surface dipoles of the fluoropolymers generated by substituting fluorine atoms with hydrogen or chlorine atoms can strongly interact with polar contacting liquids. The wettabilities of the partially fluorinated polymers were enhanced by increasing the density of dipoles across the surfaces and by introducing differentially distributed dipoles. PMID- 18393485 TI - One-, two-, and three-dimensional superstructures of gold nanorods induced by dimercaptosuccinic acid. AB - A method is described for assembling gold nanorods into one-, two-, and three dimensional superstructures. The addition of dimercaptosuccinic acid (DMSA) into the nanorod solution was found to induce self-assembly of the latter to one dimensional "tapelike", two-dimensional "sheetlike" and three-dimensional "superlattice-like" structures depending on the DMSA concentration. The assembly was found to follow a smectic structure, where the nanorod long axes are parallel to each other. The rods are spaced 8.5 +/- 0.3 nm apart in the resulting structures, which extend over several micrometers in length. Organizations perpendicular to the grid were also found. The nanorod tapes were found to bend, and they form circular assemblies as well. The assembly and morphology of the nanorod structures were characterized by transmission electron microscopy and UV vis spectroscopy. The effect of the DMSA concentration as well as the pH of the medium was also studied. On the basis of several control experiments utilizing similar molecules, charge neutralization of the nanorods by the carboxylic group of DMSA was found to be the principal reason for such an assembly, while the mercapto groups render additional stability to its structure. A mechanistic model of the assembly is proposed. This type of assembly would plausibly function as a plasmonic waveguide in potential nanodevices. PMID- 18393487 TI - Reaction of trimethylchlorosilane in spin-on Silicalite-1 zeolite film. AB - We present a study on the hydrophobization of spin-on Silicalite-1 zeolite films through silylation with trimethylchlorosilane. Microporous and micro-mesoporous Silicalite-1 films were synthesized by spin coating of suspensions of Silicalite 1 nanozeolite crystallized for different times. Ellipsometric porosimetry with toluene and water adsorbates reveals that silylation decreases the porosity and makes the films hydrophobic. The decrease in porosity depends on the exposed surface area in the pores. Water contact angle measurements confirm the hydrophobicity. Fourier transform infrared spectroscopy reveals that the trimethylsilyl groups are chemisorbed selectively on isolated silanols and less on geminal and vicinal silanols due to steric limitations. Time-of-flight secondary-ion mass spectroscopy and in situ ellipsometry analysis of the reaction kinetics show that the silylation is a bulk process occurring in the absence of diffusion limitation. Electrical current leakage on films decreases upon silylation. Silylation with trimethylchlorosilane is shown to be an effective hydrophobization method for spin-on Silicalite-1 zeolite films. PMID- 18393486 TI - Ultra-high vacuum surface analysis study of rhodopsin incorporation into supported lipid bilayers. AB - Planar supported lipid bilayers that are stable under ambient atmospheric and ultra-high-vacuum conditions were prepared by cross-linking polymerization of bis sorbylphosphatidylcholine (bis-SorbPC). X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) were employed to investigate bilayers that were cross-linked using either redox-initiated radical polymerization or ultraviolet photopolymerization. The redox method yields a more structurally intact bilayer; however, the UV method is more compatible with incorporation of transmembrane proteins. UV polymerization was therefore used to prepare cross-linked bilayers with incorporated bovine rhodopsin, a light activated, G-protein-coupled receptor (GPCR). A previous study (Subramaniam, V.; Alves, I. D.; Salgado, G. F. J.; Lau, P. W.; Wysocki, R. J.; Salamon, Z.; Tollin, G.; Hruby, V. J.; Brown, M. F.; Saavedra, S. S. J. Am. Chem. Soc. 2005, 127, 5320 5321) showed that rhodopsin retains photoactivity after incorporation into UV polymerized bis-SorbPC, but did not address how the protein is associated with the bilayer. In this study, we show that rhodopsin is retained in supported bilayers of poly(bis-SorbPC) under ultra-high-vacuum conditions, on the basis of the increase in the XPS nitrogen concentration and the presence of characteristic amino acid peaks in the ToF-SIMS data. Angle-resolved XPS data show that the protein is inserted into the bilayer, rather than adsorbed on the bilayer surface. This is the first study to demonstrate the use of ultra-high-vacuum techniques for structural studies of supported proteolipid bilayers. PMID- 18393488 TI - Pharmaceutically active natural product synthesis and supply via plant cell culture technology. AB - The chemical diversity of plant-derived natural products allows them to function in a multitude of ways including flavor enhancers, agricultural chemicals, and importantly, human medicinals. Supply of pharmaceutically active natural products is often a challenge due to the slow growing nature of some species, low yields found in nature, and unpredictable variability in accumulation. Several production options are available including natural harvestation, total chemical synthesis, semisynthesis from isolated precursors, and expression of plant pathways in microbial systems. However, for some medicinal natural products, such as the anticancer agent paclitaxel, where low yields in nature, chemical complexity and lack of knowledge of the complete biosynthetic pathway, preclude many of these options, plant cell culture technology is an attractive alternative for supply. Plant cell suspension cultures are amenable to scale-up, environmental optimization, and metabolic engineering. This review focuses on some of the key challenges in utilizing and commercializing plant cell culture suspension technology, with a focus on pharmaceutically active natural products. Recent research has been directed toward application of traditional strategies such as reactor design, cell immobilization, and enzyme elicitation as well as emerging strategies such as characterizing cellular heterogeneity and variability through flow cytometric techniques, metabolic engineering, and system-wide analysis. PMID- 18393489 TI - Synthesis of indoleamine 2,3-dioxygenase inhibitory analogues of the sponge alkaloid exiguamine A. AB - Synthetic analogues of the sponge natural product exiguamine A (3) have been prepared and evaluated for their ability to inhibit indoleamine 2,3-dioxygenase in vitro. PMID- 18393490 TI - Flexophore, a new versatile 3D pharmacophore descriptor that considers molecular flexibility. AB - A novel pharmacophore descriptor Flexophore is presented, which considers molecular flexibility when comparing descriptor similarities. The descriptor is a complete reduced graph of the underlying molecule. Its nodes are represented by enhanced MM2 atom types, while the edge descriptions encode the molecular flexibility by means of a histogram of node distances in a diverse conformer distribution. For comparing two descriptor nodes, a statistical function derived from the Cambridge Crystallographic Database is implemented. To assess the capability of the descriptor to describe the bioactivity space, 350 test data sets with 1000 molecules each are compiled. The data sets were spiked with molecules active on one of 18 different targets. In 175 of the 350 data sets, all molecules chemically similar to the query molecules were removed. Virtual screening on these data sets showed that the Flexophore descriptor detects active molecules despite chemical dissimilarity, whereas the results for the screening of the complete data sets show enrichments comparable to chemical fingerprint descriptors. The diversity analysis of the enriched compounds demonstrates that the Flexophore descriptor describes the chemical space orthogonal to chemical fingerprint descriptors. PMID- 18393491 TI - Unique structural trends in the lanthanoid oxocarbonyl complexes. AB - Reactions of laser-ablated lanthanoid atoms (except for radioactive Pm) with carbon dioxide molecules in solid argon have been investigated using matrix isolation infrared spectroscopy. On the basis of isotopic shifts, mixed isotopic splitting patterns, and CCl4-doping experiments, the lanthanoid oxocarbonyl complexes have been identified. Density functional theory calculations have been performed on these products, which support the experimental assignments of the infrared spectra. Infrared spectroscopic studies of these lanthanoid complexes combined with theoretical calculations reveal that the early lanthanoid (La-Sm) oxocarbonyl complexes adopt trans configurations, the europium and ytterbium ones adopt side-on-bonded modes (Eu-(eta2-OC)O and Yb-(eta2-OC)O), and the late lanthanoid (Gd-Lu) ones adopt cis configurations. Natural bond orbital analysis indicates that the formation of the lanthanoid oxocarbonyl complexes involves the promotion of 6s and 4f electrons into the metal valence shell. PMID- 18393492 TI - Insights into DNA binding of ruthenium arene complexes: role of hydrogen bonding and pi stacking. AB - Density functional theory (DFT) methods are used to investigate the binding of ruthenium arene complexes, proposed as promising anticancer drugs, to isolated nucleobases. This shows a clear preference for binding at guanine over any other base and an approximately 100 kJ mol (-1) difference in binding between guanine and adenine in the gas phase, while binding to cytosine and inosine are intermediate in energy between these extremes. Solvation reduces binding energies and the discrimination between bases but maintains the overall pattern of binding. DFT and ab initio data on arene-base interactions in the absence of ruthenium show that stacking and hydrogen-bonding interactions play a significant role but cannot account for all of the energy difference between bases observed. Atoms-in-molecules analysis allows further decomposition of binding energies into contributions from covalent-binding, hydrogen-bonding, and pi-stacking interactions. Larger arenes undergo stabilizing stacking interactions, whereas N H...X hydrogen bonding is independent of arene. Pairing of guanine to cytosine is affected by ruthenium complexation, with individual hydrogen-bonding energies being altered but the overall pairing energy remaining almost constant. PMID- 18393493 TI - Three-dimensional homochiral transition-metal camphorate architectures directed by a flexible auxiliary ligand. AB - Reported here are four homochiral and one racemic chiral compounds assembled from transition metals (Fe2+, Co2+, and Ni2+), rigid enantiopure camphoric acid, and flexible 4,4'-trimethylenedipyridine; they feature an unusual homochiral three dimensional supramolecular assembly (resulting from catenation of 4-connected bilayers) as well as a homochiral 3-fold interpenetrating diamond net and a noninterpenetrating primitive cubic net. PMID- 18393494 TI - Zigzag boron-carbon nanotubes with quasi-planar tetracoordinate carbons. AB - Four kinds of novel zigzag boron-carbon nanotubes with quasi-planar tetracoordinate carbons (2m, 0, i) (m = 3-6; i = 1-4) have been constructed, and their structures, stabilities, and bonding properties have been investigated by B3LYP calculations. The results show that the novel nanotubes (m, 0, i) have the character of metal properties with quite small HOM-LUMO gaps. Structurally, there are big windows on the walls of nanotubes and they are promising in design for functional materials. PMID- 18393496 TI - Synthesis and AFM structural imaging of dendrimer-like star-branched polystyrenes. AB - We report on the synthesis of large and densely tough dendrimer-like star branched polystyrenes of generation 3, specially designed for their visualization by AFM imaging. For this purpose, an iterative methodology involving a sequential three stages synthetic approach has been adopted. The precisely designed G-3 polymers appear on AFM images as completely flattened objects with a symmetrical square-like shape and an average diameter of 250 nm and a height of less than 1 nm. Imaging of their internal structure indicates that they are constituted of four interconnected lobes, in relation with the use of a tetrafunctional core precursor. In addition, AFM images show the presence inside each object of all the branching points which appear as white harder zones regularly distributed in the less dense and softer PS matrix. PMID- 18393495 TI - A polymeric domain that promotes cellular internalization. AB - Polymers have emerged as powerful biological tools; however, their ability to gain access to the intracellular environment is limited. To expand the biological utility of polymer scaffolds, we have synthesized an internalization domain using the ring-opening metathesis polymerization (ROMP). A polymer functionalized with guanidinium groups is effectively internalized by cells and localized in both vesicles and the cytoplasm. Because the synthesis of such materials is modular, we anticipate that compounds of this type can be fashioned that facilitate the delivery of cargo via end-cap derivatization or block copolymer synthesis. PMID- 18393497 TI - Photoinduced ultrafast electron transfer from CdSe quantum dots to Re-bipyridyl complexes. AB - Ultrafast dissociation of excitons in CdSe quantum dots via electron transfer to adsorbed Re-bipyridyl complexes was demonstrated. The dissociation pathway was determined by the observation of reduced adsorbate using femtosecond IR spectroscopy. The rate of electron transfer was shown to increase at smaller QD sizes. Electron transfer time as fast as 2.3 ps was observed, faster than the exciton annihilation time in CdSe. The ultrafast charge separation in this quantum dot-adsorbate donor-acceptor complex provides a potential approach for separating multiple excitons in quantum dots. PMID- 18393498 TI - Organocatalysis in a synthetic receptor with an inwardly directed carboxylic acid. AB - A cavitand functionalized with a Kemp's triacid derivative was used to catalyze the epoxide ring-opening cyclizations of 1,5-epoxyalcohols. A deep, cylindrical cavity containing electron-rich aromatic walls and an inwardly directed carboxylic acid displayed the necessary characteristics to bind different 1,5 epoxyalcohols and initiate their cyclization reactions. The reactions inside this synthetic receptor occurred in a catalytic and regioselective manner. These results highlight that the arrangement of functionality and unique solvation provided by the structured interiors of natural enzymes can be incorporated into synthetic systems having useful physical and chemical properties. PMID- 18393499 TI - Studies on a new biosynthetic pathway for menaquinone. AB - Menaquinone is biosynthesized from chorismate via a new pathway involving 1,4 dihydroxy-6-naphthoate in Streptomyces and presumably in pathogenic bacteria Helicobacter and Campylobacter. PMID- 18393500 TI - Nanoparticle MALDI-TOF mass spectrometry without fragmentation: Au25(SCH2CH2Ph)18 and mixed monolayer Au25(SCH2CH2Ph)(18-x)(L)(x). AB - Intact molecular ions of the organothiolate-protected nanoparticle Au25(SCH2CH2Ph)18, including their isotopic resolution, can be observed at 7391 Da as 1- and 1+ ions in negative and positive mode, respectively, by MALDI-TOF mass spectrometry when using a tactic of threshold laser pulse intensities and trans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene]malononitrile (DCTB) as matrix. Previous MALDI-TOF studies of Au nanoparticles using other matrices have encountered extensive fragmentation of nanoparticle as well as thiolate ligands. Absence of fragmentation enables precise determination of the distribution of mixed monolayer compositions on nanoparticles prepared by ligand exchange reactions and by synthesis using thiol mixtures. Reaction conditions producing mixed monolayers containing only one or a small number of usefully functional ligands can be readily identified. At increased laser pulse intensity, the first fragmentation step(s) for the Au25(SCH 2CH2Ph)18 nanoparticle results in losses of AuL units and, in particular, loss of Au4(SCH2CH2Ph)4. PMID- 18393501 TI - Interactions of the hormone oxytocin with divalent metal ions. AB - The interaction of the cyclic nonapeptide oxytocin (OT) with a number of alkaline earth and divalent transition metal ions (X(2+)) was examined employing mass spectrometry (MS) and ion mobility spectrometry (IMS) techniques in combination with molecular dynamics (MD) and density functional theory (DFT) calculations. Under acidic conditions it was found that OT exhibits an exceptionally strong affinity for all divalent metal ions resulting in strong [OT + X](2+) peaks in the mass spectrum. Under basic conditions only Cu(2+) and Ni(2+)-OT complexes were detected and these were singly, doubly, triply, or quadruply deprotonated. Collision-induced dissociation of the [OT - 3H + Cu](-) complex yielded exclusively C-terminal Cu(2+)-containing fragments (Cu(2+)fragment(3-)), suggesting that the Cu(2+) ligation site includes deprotonated C-terminal backbone amide nitrogen atoms and the N-terminal amino nitrogen atom in [OT - 3H + Cu](-). MD and DFT calculations indicate a square-planar complex is consistent with these observations and with experimental collision cross sections. MD and DFT calculations also indicate either an octahedral or trigonal-bipyramidal complex between Zn(2+) and OT is lowest in energy with carbonyl oxygens being the primary ligation sites. Both complexes yield cross sections in agreement with experiment. The biological impact of the structural changes induced in OT by divalent metal ion coodination is discussed. PMID- 18393502 TI - Optofluidic encapsulation of crystalline colloidal arrays into spherical membrane. AB - Double emulsion droplets encapsulating crystalline colloidal arrays (CCAs) with a narrow size distribution were produced using an optofluidic device. The shell phase of the double emulsion was a photocurable resin that was photopolymerized downstream of the fluidic channel within 1 s after drop generation. The present optofluidic synthesis scheme was very effective for fabricating highly monodisperse spherical CCAs that were made structurally stable by in situ photopolymerization of the encapsulating shells. The shell thickness and the number of core emulsion drops could be controlled by varying the flow rates of the three coflowing streams in the dripping regime. The spherical CCAs confined in the shell exhibited distinct diffraction patterns in the visible range, in contrast to conventional film-type CCAs. As a result of their structure, the spherical CCAs exhibited photonic band gaps for normal incident light independent of the position on the spherical surface. This property was induced by heterogeneous nucleation at the smooth wall of the spherical emulsion drop during crystallization into a face-centered cubic (fcc) structure. On the other hand, the solidified shells did not permit the penetration of ionic species, enabling the CCAs to maintain their structure in a continuous aqueous phase of high ionic strength for at least 1 month. In addition, the evaporation of water molecules inside the shell was slowed considerably when the core-shell microparticles were exposed to air: It took approximately 6 h for a suspension encapsulated in a thick shell to evaporate completely, which is approximately 1000 times longer than the evaporation time for water droplets with the same volume. Finally, the spherical CCAs additionally exhibited enhanced stability against external electric fields. The spherical geometry and high dielectric constant of the suspension contributed to reducing the electric field inside the shell, thereby inhibiting the electrophoretic movement of the charged particles. PMID- 18393503 TI - EP-2104R: a fibrin-specific gadolinium-Based MRI contrast agent for detection of thrombus. AB - Thrombus (blood clot) is implicated in a number of life threatening diseases, e.g., heart attack, stroke, pulmonary embolism. EP-2104R is an MRI contrast agent designed to detect thrombus by binding to the protein fibrin, present in all thrombi. EP-2104R comprises an 11 amino acid peptide derivatized with 2 GdDOTA like moieties at both the C- and N-terminus of the peptide (4 Gd in total). EP 2104R was synthesized by a mixture of solid phase and solution techniques. The La(III) analogue was characterized by and 1D and 2D NMR spectroscopy and was found to have the expected structure. EP-2104R was found to be significantly more inert to Gd(III) loss than commercial contrast agents. At the most extreme conditions tested (pH 3, 60 degrees C, 96 hrs), less than 10% of Gd was removed from EP-2104R by a challenge with a DTPA based ligand, while the commercial contrast agents equilibrated within minutes to hours. EP-2104R binds equally to two sites on human fibrin (Kd = 1.7 +/- 0.5 microM) and has a similar affinity to mouse, rat, rabbit, pig, and dog fibrin. EP-2104R has excellent specificity for fibrin over fibrinogen (over 100-fold) and for fibrin over serum albumin (over 1000-fold). The relaxivity of EP-2104R bound to fibrin at 37 degrees C and 1.4 T was 71.4 mM(-1) s(-1) per molecule of EP-2104R (17.4 per Gd), about 25 times higher than that of GdDOTA measured under the same conditions. Strong fibrin binding, fibrin selectivity, and high molecular relaxivity enable EP-2104R to detect blood clots in vivo. PMID- 18393504 TI - Cooperative, highly enantioselective phosphinothiourea catalysis of imine-allene [3 + 2] cycloadditions. AB - A new family of phosphinthiourea catalysts was developed for the highly enantioselective synthesis of 2-aryl-2,5-hydropyrroles via a [3 + 2] cycloaddition of an electron-deficient allene with aryl and heteroaryl diphenylphosphinoylimines. The presence of both H2O and Et3N as additives was found to be important for achieving optimal rates. Dual activation of both nucleophile and electrophile by the bifunctional catalyst is invoked to account for the observed high reactivity and enantioselectivity. PMID- 18393505 TI - Magic angle spinning NMR experiments for structural studies of differentially enriched protein interfaces and protein assemblies. AB - Protein-protein interactions play vital roles in numerous biological processes. These interactions often result in formation of insoluble and noncrystalline protein assemblies. Solid-state NMR spectroscopy is rapidly emerging as a premier method for structural analysis of such systems. We introduce a family of two dimensional magic angle spinning (MAS) NMR experiments for structural studies of differentially isotopically enriched protein assemblies. Using 1 73((13)C,(15)N)/74-108((15)N) labeled thioredoxin reassembly, we demonstrate that dipolar dephasing followed by proton-assisted heteronuclear magnetization transfer yields long-range (15)N-(13)C correlations arising exclusively from the interfaces formed by the pair of differentially enriched complementary fragments of thioredoxin. Incorporation of dipolar dephasing into the (15)N proton-driven spin diffusion and into the (1)H-(15)N FSLG-HETCOR sequences permits (1)H and (15)N resonance assignments of the 74-108((15)N) enriched C-terminal fragment of thioredoxin alone. The differential isotopic labeling scheme and the NMR experiments demonstrated here allow for structural analysis of both the interface and each interacting protein. Isotope editing of the magnetization transfers results in spectral simplification, and therefore larger protein assemblies are expected to be amenable to these experiments. PMID- 18393506 TI - Oxidation of porcine Myosin by hypervalent myoglobin: the role of thiol groups. AB - Oxidation of the myofibrillar muscle protein myosin from pork by hypervalent myoglobin species (MbFe(III)/H 2O2 radical generating system) was investigated in aqueous solution in the pH range of 5.0-7.8 by electron spin resonance (ESR) spectroscopy using N- tert-butyl-alpha-phenylnitrone (PBN) as spin trap and indirectly by determination of the rate of reduction of hypervalent myoglobin species by UV spectroscopy. Cross-linking of myosin was examined by SDS-PAGE. The target for oxidative modification of myosin was studied by thiol blocking by N acetylmaleimide (NEM) and by determining oxidative modification of myosin thiols. The reaction between myosin and hypervalent myoglobin was fast and showed little dependence on pH. The myosin radicals formed were observed to be short-lived. Myosin thiols are suggested to be the main target for oxidative modification, as NEM-treated myosin did not form radicals in the presence of hypervalent myoglobin. A significant decrease in thiol content was already demonstrated 25 s after initiation of oxidation of myosin. The majority of myosin heavy chain (MHC) was demonstrated to be cross-linked through intermolecular disulfide bonding 1 h after initiation of oxidation. This demonstrates that thiols are important for radical formation and cross-linking of myosin during oxidation with hypervalent myoglobin at the pH of meat products. PMID- 18393507 TI - Synthetic studies on maitotoxin. 2. Stereoselective synthesis of the WXYZA'-ring system. AB - The stereoselective synthesis of the WXYZA'-ring system of maitotoxin has been accomplished via a linear synthetic approach, in which key reactions were SmI 2 induced cyclization of beta-alkoxyacrylate for the construction of the A'-, Y-, and X-rings and 6- endo cyclization of hydroxy vinylepoxide for that of the Z- and W-rings. PMID- 18393508 TI - Bronsted-acid-catalyzed activation of nitroalkanes: a direct enantioselective aza Henry reaction. AB - A direct asymmetric organocatalytic aza-Henry reaction has been developed in which a new bifunctional Bronsted-acid-catalyzed activation of nitroalkanes provides an efficient access to alpha,beta-diamino acids with high dia- and enantioselectivities under mild and base-free reaction conditions. PMID- 18393509 TI - Synthetic studies on maitotoxin. 3. Stereoselective synthesis of the BCDE-ring system. AB - The stereoselective synthesis of the BCDE-ring system of maitotoxin has been accomplished through a two-directional strategy for the construction of polycyclic ether. The key reactions involve SmI 2-induced double cyclization of a beta-alkoxyacrylate and a double dihydroxylation for construction of the B- and E rings. PMID- 18393510 TI - Synthetic studies on maitotoxin. 1. Stereoselective synthesis of the C'D'E'F' ring system having a side chain. AB - The stereoselective synthesis of the maitotoxin C'D'E'F'-ring system having a side chain has been accomplished through a convergent strategy. The key reactions include Horner-Wadsworth-Emmons coupling of the C'D'E'-ring and the side chain and subsequent construction of the F'-ring by silane reduction of dihydropyran. PMID- 18393511 TI - Structure and semisynthesis of platensimide A, produced by Streptomyces platensis. AB - Platensimycin and platencin are novel natural product antibiotics that inhibit bacterial growth by inhibiting condensing enzymes FabF and FabF/FabH of fatty acid biosynthesis pathways, respectively. Continued search for the natural congeners of these compounds led to the isolation of platensic acid, the free C 17 tetracyclic enoic acid, and platensimide A, a 2,4-diaminobutyric acid amide derivative. Isolation, structure, semisynthesis, and activity of these compounds are described. PMID- 18393512 TI - One-pot synthesis of nucleoside 5'-triphosphates from nucleoside 5'-H phosphonates. AB - Nucleoside 5'-triphosphates (NTPs) play key roles in biology and medicine. However, these compounds are notoriously difficult to synthesize. We describe a one-pot method to prepare NTPs from nucleoside 5'-H-phosphonate monoesters via pyridinium phosphoramidates, and we used this approach to synthesize ATP, UTP, GTP, CTP, ribavirin-TP, and 6-methylpurine ribonucleoside-TP (6MePTP). Poliovirus RNA-dependent RNA polymerase efficiently employed 6MePTP as a substrate, suggesting that the cognate nucleoside, a poorly understood antiviral agent, may damage viral RNA. PMID- 18393513 TI - Oxidation of a biomarker for phenol carcinogen exposure: expanding the redox chemistry of 2'-deoxyguanosine. AB - A biomarker for phenolic carcinogen exposure, 8-(4''-hydroxyphenyl)-2' deoxyguanosine, has been found to undergo oxidative coupling in the presence of Na2IrCl6 to afford ortho-ortho C-C-coupled polyphenols through the intermediacy of a phenoxyl radical. One can envision using such unique chemistry to oxidatively couple strands of DNA for the generation of new biomaterials. Our results also demonstrate the potential for phenolic adducts of DNA to undergo further oxidation reactions that may contribute to phenol-mediated cytotoxicity and genotoxicity. PMID- 18393514 TI - Synthesis of (2-oxoindolin-3-ylidene)methyl acetates involving a C-H functionalization process. AB - A novel palladium-catalyzed oxidative C-H functionalization protocol for the synthesis of (2-oxoindolin-3-ylidene)methyl acetates has been developed. In the presence of Pd(OAc)2 and PhI(OAc)2, a variety of N-arylpropiolamides underwent the C-H functionalization reaction with acids to afford the corresponding (E)-(2 oxoindolin-3-ylidene)methyl acetates selectively in moderate to excellent yields. PMID- 18393515 TI - Pd-catalyzed carboetherification of beta,gamma-unsaturated oximes: a novel approach to Delta2-isoxazolines. AB - A novel route to the synthesis of Delta2-isoxazoline derivatives is described. Reaction of beta,gamma-unsaturated oximes with aryl bromides via palladium catalyzed carboetherification affords 3,5-disubstituted Delta2-isoxazolines in good yields. The use of Xantphos as ligand is crucial for the transformation. The carboetherification products can be further converted to beta-hydroxy ketones in the presence of Fe powder and NH4Cl. PMID- 18393516 TI - Diastereoselective Brook rearrangement-mediated [3+4] annulation: application to a formal synthesis of (+)-laurallene. AB - The formal synthesis of (+)-laurallene, a halogenated eight-membered ring ether, was accomplished. The synthesis involves construction of a trans alpha,alpha' disubstituted oxocene structure 16 through a Brook rearrangement-mediated [3+4] annulation using acryloylsilane 10 and 6-oxa-2-cycloheptenone 9 and its conversion into 2, which has been transformed into (+)-laurallene by Crimmins and co-workers. PMID- 18393517 TI - Asymmetric direct aldol reactions of pyruvic derivatives. AB - Simple chiral primary-tertiary diamine-Bronsted acid conjugates such as 1e can effectively catalyze the direct aldol reactions of pyruvic derivatives with excellent syn diastereoselectivities and enantioselectivities, thus functionally mimicking the pyruvate-dependent type I aldolases. PMID- 18393518 TI - Expedient synthesis of the tetracyclic core of ent-nakadomarin A. AB - An efficient eight-step assembly of the tetracyclic core (ABCD rings) of ent-(+) nakadomarin A, a bioactive hexacyclic marine alkaloid, has been realized with Sonogashira coupling, platinum(II)-promoted cascade cyclizations, and saturation of a challenging carbon-carbon double bond through a hydroboration/oxidation/xanthate formation/Barton-McCombie deoxygenation sequence as key transformations. PMID- 18393519 TI - Mo-Au combo catalysis for rapid 1,3-rearrangement of propargyl alcohols into alpha,beta-unsaturated carbonyl compounds. AB - The combination of Mo and cationic Au catalysts dramatically accelerated the rearrangement of diverse propargyl alcohols, which includes a short reaction time, mild conditions, and high product yields. A practical application to the highly challenging primary propargyl alcohols and the N-alkynyl amides is achieved. PMID- 18393520 TI - Application of a domino Friedel-Crafts acylation/alkylation reaction to the formal syntheses of (+/-)-taiwaniaquinol B and (+/-)-dichroanone. AB - An efficient acid-promoted domino Friedel-Crafts (FC) acylation/alkylation reaction has been developed for the construction of the core 6,5,6-ABC tricyclic skeleton of taiwaniaquinoids. The formal total syntheses of diterpenoids (+/-) taiwaniaquinol B and (+/-)-dichroanone based on this strategy have been achieved. PMID- 18393521 TI - Palladium-catalyzed sp(3) CH activation of simple alkyl groups: direct preparation of indoline derivatives from N-alkyl-2-bromoanilines. AB - The sp3 C-H activation of a simple alkyl group catalyzed by palladium(0) provides a novel and convenient strategy for the synthesis of various indolines from simple precursors, such as N-alkyl-2-bromoanilines. This study demonstrates that assisting moieties in the substrate such as a pyridine or quaternary carbon are not always necessary for sp3 C-H activation. PMID- 18393522 TI - Suzuki-Miyaura cross-coupling of potassium trifluoroboratohomoenolates. AB - Ketone-, ester-, and amide-containing potassium trifluoroboratohomoenolates were prepared in good to excellent yields from the corresponding unsaturated carbonyl compounds. They were shown to be effective coupling partners in the Suzuki Miyaura reaction with a variety of electrophiles including electron-rich and electron-poor aryl bromides and -chlorides. PMID- 18393523 TI - Interactions of bovine serum albumin with ethylene oxide/butylene oxide copolymers in aqueous solution. AB - The interactions of bovine serum albumin (BSA) with three ethylene oxide/butylene oxide (E/B) copolymers having different block lengths and varying molecular architectures is examined in this study in aqueous solutions. Dynamic light scattering (DLS) indicates the absence of BSA-polymer binding in micellar systems of copolymers with lengthy hydrophilic blocks. On the contrary, stable protein polymer aggregates were observed in the case of E 18B 10 block copolymer. Results from DLS and SAXS suggest the dissociation of E/B copolymer micelles in the presence of protein and the absorption of polymer chains to BSA surface. At high protein loadings, bound BSA adopts a more compact conformation in solution. The secondary structure of the protein remains essentially unaffected even at high polymer concentrations. Raman spectroscopy was used to give insight to the configurations of the bound molecules in concentrated solutions. In the vicinity of the critical gel concentration of E 18B 10 introduction of BSA can dramatically modify the phase diagram, inducing a gel-sol-gel transition. The overall picture of the interaction diagram of the E 18B 10-BSA reflects the shrinkage of the suspended particles due to destabilization of micelles induced by BSA and the gelator nature of the globular protein. SAXS and rheology were used to further characterize the structure and flow behavior of the polymer protein hybrid gels and sols. PMID- 18393524 TI - Efficient approach to design stable water-dispersible nanoparticles of hydrophobic cellulose esters. AB - Commercially prepared cellulose acetate, cellulose acetate propionate, -butyrate, and -phthalate as well as cellulose acetates prepared in the laboratory scale with varying degree of substitution (DS) self-assemble into regular nanoparticles, ranging in size from 86 to 368 nm, by using two different techniques of nanoprecipitation. Dialysis of polymers dissolved in N,N dimethylacetamide results in the formation of regular nanospheres whereas the preparation in acetone by successive adding of water leads to bean-shaped particles in the nanoscale. One criterion for nanoprecipitation is the existence of dilute polymer solutions. Furthermore, the formation of nanoparticles strongly depends on DS and distribution of the substituents. Concerning this issue, quantitative (13)C NMR spectroscopy was applied for detailed structure characterization of selected cellulose acetates. The stability of the nanoparticle suspensions in the physiological pH range was observed by zeta potential measurements. PMID- 18393525 TI - Construction of the active site of glutathione peroxidase on polymer-based nanoparticles. AB - A new nanoenzyme model with glutathione peroxidase-like active site was constructed on polystyrene nanoparticle (PN1) via microemulsion polymerization. In this model system, two functional monomers were designed: one is a tellurium containing compound that was introduced on the surface of the nanoparticle and acts as a catalytic center, and the other one is an arginine-containing compound designed as a binding site for the complexation of the carboxyl group of substrate 3-carboxy-4-nitrobenzenethiol (ArSH, 1). As a new glutathione peroxidase (GPx) mimic, it demonstrated excellent catalytic activity and substrate specificity. In ArSH assay system, it was at least 316,000-fold more efficient than PhSeSePh for the reduction of cumene hydroperoxide (CUOOH) by ArSH. In contrast to model PN2, which lacks of substrate binding site, PN1 exhibits an obvious enhancement in catalytic activity. To further promote the catalytic efficiency, a substrate ArSH surface-imprinted nanoenzyme model (I-PN) was developed. By correctly incorporating and positioning the catalytic center tellurium and functional binding factor guanidinium, a continuative activity enhancement of 596,000-fold for the reduction of CUOOH by catalyst I-PN compared with diphenyl diselenide (PhSeSePh) was observed. The results clearly show that polymeric nanoparticle can be developed as an excellent model for combining most of catalytic factors of enzyme into one scaffold. PMID- 18393527 TI - An easy microwave-assisted synthesis of sulfonamides directly from sulfonic acids. AB - An easy and handy synthesis of sulfonamides directly from sulfonic acids or its sodium salts is reported. The reaction is performed under microwave irradiation, has shown a good functional group tolerance, and is high yielding. PMID- 18393528 TI - Toward multistation rotaxanes using metalloporphyrin coordination templating. AB - This paper describes some approaches toward the templated synthesis of rotaxanes incorporating strapped metalloporphyrin moieties as the shuttle unit, with the thread component containing both a neutral diimide "station" and a functionalized pyridine moiety, the latter acting not only as a template but also as a second binding motif. In the first instance, the use of appropriately 3,5 difunctionalized pyridine esters and naphthoquinol-strapped rhodium(III) chloride porphyrins in a stoppering approach to rotaxanes produced only unlinked components: the flexibility of the strap allowed sufficient room for the potential thread unit to bind on the same face of the porphyrin as the strap, while not being interlocked through it. An alternative strategy involving a 1,3 dipolar cycloaddition reaction (a "click" reaction) between azides and alkynes, producing triazole linkers in the thread component of rotaxanes, was more successful. Both porphyrinic (zinc, free base, and rhodium(III) derivatives) and crown ether rotaxanes were successfully produced, with multifunctional (triazole and naphthodiimide) thread units. The potential for molecular motion through the use of stimuli such as acid, solvent, and competing ligands was investigated, with limited success. The same cycloaddition methodology was extended to pyridine templated analogues of the thread components in the Rh(III)-strapped porphyrins, but again, only unlinked thread and porphyrin shuttle units were produced. PMID- 18393529 TI - Formation of supramolecular polymers and discrete dimers of perylene bisimide dyes based on melamine-cyanurates hydrogen-bonding interactions. AB - Melamine-linked perylene bisimide dyes (MPBIs) bearing an ethylene or trimethylene group as linker moieties were synthesized, and their self aggregation and coaggregation with cyanurates through complementary triple hydrogen bonds have been investigated. UV/vis studies revealed that both the MPBIs self-assemble in nonpolar organic solvent through pi-pi stacking interaction between perylene cores, giving self-aggregates with nearly identical thermal stabilities. Upon addition of 1 equiv of cyanurate components, however, the stabilities of the resulting aggregates were dramatically changed between the two systems, suggesting the formation of different types of hydrogen-bonded supramolecular species. Dynamic light scattering and atomic force microscopic studies revealed that the system featuring ethylene linker moieties generates a discrete dimer of MPBI supported by two cyanurate molecules, whereas the system featuring trimethylene linker moieties affords extended supramolecular polymers hierarchically organizing into nanoscopic fibers. These results demonstrate that it is possible to obtain distinct supramolecular species by just changing the number of carbon atoms at the linker moieties of MPBI components. The present strategy for the fabrication of discrete or polymeric supramolecular assemblies should be applicable to other functional pi-conjugated molecules. PMID- 18393530 TI - Real-time microfluidic system for studying mammalian cells in 3D microenvironments. AB - We describe a microfluidic system that can control, in real time, the microenvironments of mammalian cells in naturally derived 3D extracellular matrix (ECM). This chip combines pneumatically actuated valves with an individually addressable array of 3D cell-laden ECM; actuation of valves determines the pathways for delivering reagents through the chip and for exchanging diffusible factors between cell chambers. To promote rapid perfusion of reagents through 3D gels (with complete exchange of reagents within the gel in seconds), we created conduits above the gels for fluid flow, and microposts to stabilize the gels under high perfusion rates. As a biological demonstration, we studied spatially segregated mouse embryonic stem cells and mouse embryonic fibroblasts embedded in 3D Matrigel over days of culture. Overall, this system may be useful for high throughput screening, single-cell analysis and studies of cell-cell communication, where rapid control of 3D cellular microenvironments is desired. PMID- 18393531 TI - Quantitative analysis of HIV-1 protease inhibitors in cell lysates using MALDI FTICR mass spectrometry. AB - In this report we explore the use of MALDI-FTICR mass spectrometry for the quantitative analysis of five HIV-1 protease inhibitors in cell lysates. 2,5 Dihydroxybenzoic acid (DHB) was used as the matrix. From a quantitative perspective, DHB is usually a poor matrix due to its poor shot-to-shot and poor spot-to-spot reproducibilities. We found that the quantitative precisions improved significantly when DMSO (dimethylsulfoxide) was added to the matrix solution. For lopinavir and ritonavir, currently the most frequently prescribed HIV-1 protease inhibitors, the signal-to-noise ratios improved significantly when potassium iodide was added to the matrix solution. The mean quantitative precisions, expressed as % relative standard deviation, were 6.4% for saquinavir, 7.3% for lopinavir, 8.5% for ritonavir, 11.1% for indinavir, and 7.2% for nelfinavir. The mean quantitative accuracies, expressed as % deviation, were 4.5% for saquinavir, 6.0% for lopinavir, 5.9% for ritonavir, 6.6% for indinavir, and 8.0% for nelfinavir. The concentrations measured for the individual quality control samples were all within 85-117% of the theoretical concentrations. The lower limits of quantification in cell lysates were 4 fmol/microL for saquinavir, 16 fmol/microL for lopinavir, 31 fmol/microL for ritonavir, and 100 fmol/microL for indinavir and nelfinavir. The mean mass accuracies for the protease inhibitors were 0.28 ppm using external calibration. Our results show that MALDI FTICR mass spectrometry can be successfully used for precise, accurate, and selective quantitative analyses of HIV-1 protease inhibitors in cell lysates. In addition, the lower limits of quantification obtained allow clinical applications of the technique. PMID- 18393532 TI - Domain stabilities in protein kinase R (PKR): evidence for weak interdomain interactions. AB - PKR (protein kinase R) is induced by interferon and is a key component of the innate immunity antiviral pathway. Upon binding dsRNA, PKR undergoes autophosphorylation reactions that activate the kinase, leading it to phosphorylate eIF2alpha, thus inhibiting protein synthesis in virally infected cells. PKR contains a dsRNA-binding domain (dsRBD) and a kinase domain. The dsRBD is composed of two tandem dsRNA-binding motifs. An autoinhibition model for PKR has been proposed, whereby dsRNA binding activates the enzyme by inducing a conformational change that relieves the latent enzyme of the inhibition that is mediated by the interaction of the dsRBD with the kinase. However, recent biophysical data support an open conformation for the latent enzyme, where activation is mediated by dimerization of PKR induced upon binding dsRNA. We have probed the importance of interdomain contacts by comparing the relative stabilities of isolated domains with the same domain in the context of the intact enzyme using equilibrium chemical denaturation experiments. The two dsRNA-binding motifs fold independently, with the C-terminal motif exhibiting greater stability. The kinase domain is stabilized by about 1.5 kcal/mol in the context of the holenzyme, and we detect low-affinity binding of the kinase and dsRBD constructs in solution, indicating that these domains interact weakly. Limited proteolysis measurements confirm the expected domain boundaries and reveal that the activation loop in the kinase is accessible to cleavage and unstructured. Autophosphorylation induces a conformation change that blocks proteolysis of the activation loop. PMID- 18393533 TI - Role of ribosomal protein L27 in peptidyl transfer. AB - The current view of ribosomal peptidyl transfer is that the ribosome is a ribozyme and that ribosomal proteins are not involved in catalysis of the chemical reaction. This view is largely based on the first crystal structures of bacterial large ribosomal subunits that did not show any protein components near the peptidyl transferase center (PTC). Recent crystallographic data on the full 70S ribosome from Thermus thermophilus, however, show that ribosomal protein L27 extends with its N-terminus into the PTC in accordance with independent biochemical data, thus raising the question of whether the ribozyme picture is strictly valid. We have carried out extensive computer simulations of the peptidyl transfer reaction in the T. thermophilus ribosome to address the role of L27. The results show a reaction rate similar to that obtained in earlier simulations of the Haloarcula marismortui reaction. Furthermore, deletion of L27 is predicted to only give a minor rate reduction, in agreement with biochemical data, suggesting that the ribozyme view is indeed valid. The N-terminus of L27 is predicted to interact with the A76 phosphate group of the A-site tRNA, thereby explaining the observed impairment of A-site substrate binding for ribosomes lacking L27. Simulations are also reported for the reaction with puromycin, an A site tRNA analogue which lacks the A76 phosphate group. The calculated energetics shows that this substrate can cause a downward p K a shift of L27 and that the reaction proceeds faster with the L27 N-terminus deprotonated, in contrast to the situation with aminoacyl-tRNA substrates. These results could explain the observed differences in pH dependence between the puromycin and C-puromycin reactions, where the former reaction has been seen to depend on an additional ionizing group besides the attacking amine, and our model predicts this ionizing group to be the N-terminal amine of L27. PMID- 18393534 TI - Volume and free energy of folding for troponin C C-domain: linkage to ion binding and N-domain interaction. AB - Troponin C (TnC) is an 18-kDa acidic protein of the EF-hand family that serves as the trigger for muscle contraction. In this study, we investigated the thermodynamic stability of the C-domain of TnC in all its occupancy states (apo, Mg (2+)-, and Ca (2+)-bound states) using a fluorescent mutant with Phe 105 replaced by Trp (F105W/C-domain, residues 88-162) and (1)H NMR spectroscopy. High hydrostatic pressure was employed as a perturbing agent, in combination with urea or without it. On the basis of changes in Trp emission, the C-domain apo state was denatured by pressure (in the range of 1-1000 bar) in the absence of urea. The fluorescence data were corroborated by following the changes in the (1)H NMR signal of Histidine 128. Addition of Ca (2+) or Mg (2+) increased the C-domain stability so that complete denaturation was attained only by the combined use of high hydrostatic pressure and either 7-8 M or 1.5-2 M urea, respectively. The (1)H NMR spectra in the presence of Ca (2+) was typical of a highly structured protein and allowed us to follow the changes in the local environment of several amino-acid residues as a function of pressure at 4 M Urea. Different residues presented different volume changes, but those that are in the hydrophobic core portrayed values very similar to that obtained for tryptophan 105 as measured by fluorescence, indicating that it is indeed a good probe for the overall tertiary structure. From these experiments, we calculated the thermodynamic parameters (Delta G degrees atm and Delta V) that govern the folding of the C-domain in all its possible physiological states and constructed a thermodynamic cycle. Furthermore, a comparison of the volume and free-energy changes of folding of isolated C-domain with those of intact TnC (F105W) revealed that the N-domain has little effect on the structure of the C-domain, even in the presence of Ca (2+). The volume and free-energy diagrams reveal a landscape of different conformations from the less structured, denatured apo form to the highly structured, Ca (2+) bound form. The large change in folding free energy of the C-domain that takes place when Ca (2+) binds may explain the much higher Ca (2+) affinity of sites III and IV, 2 orders of magnitude higher than the affinity of sites I and II. PMID- 18393535 TI - Comparative kinetics of cofactor association and dissociation for the human and trypanosomal S-adenosylhomocysteine hydrolases. 2. The role of helix 18 stability. AB - The S-adenosyl- l-homocysteine (AdoHcy) hydrolases (SAHH) from Homo sapiens (Hs SAHH) and from the parasite Trypanosoma cruzi (Tc-SAHH) are very similar in structure and catalytic properties but differ in the kinetics and thermodynamics of association and dissociation of the cofactor NAD (+). The binding of NAD (+) and NADH in SAHH appears structurally to be mediated by helix 18, formed by seven residues near the C-terminus of the adjacent subunit. Helix-propensity estimates indicate decreasing stability of helix 18 in the order Hs-SAHH > Tc-SAHH > Ld SAHH (from Leishmania donovani) > Pf-SAHH (from Plasmodium falciparum), which would be consistent with the previous observations. Here we report the properties of Hs-18Pf-SAHH, the human enzyme with plasmodial helix 18, and Tc-18Hs-SAHH, the trypanosomal enzyme with human helix 18. Hs-18Tc-SAHH, the human enzyme with trypanosomal helix 18, was also prepared but differed insignificantly from Hs SAHH. Association of NAD (+) with Hs-SAHH, Hs-18Pf-SAHH, Tc-18Hs-SAHH, and Tc SAHH exhibited biphasic kinetics for all enzymes. A thermal maximum in rate, attributed to the onset of local structural alterations in or near the binding site, occurred at 35, 33, 30, and 15 degrees C, respectively. This order is consistent with some reversible changes within helix 18 but does require influence of other properties of the "host enzyme". Dissociation of NAD (+) from the same series of enzymes also exhibited biphasic kinetics with a transition to faster rates (a larger entropy of activation more than compensates for a larger enthalpy of activation) at temperatures of 41, 38, 36, and 29 degrees C, respectively. This order is also consistent with changes in helix 18 but again requiring influence of other properties of the "host enzyme". Global unfolding of all fully reconstituted holoenzymes occurred around 63 degrees C, confirming that the kinetic transition temperatures did not arise from a major disruption of the protein structure. PMID- 18393536 TI - Two-dimensional center-of-mass diffusion of lipid-tethered poly(2-methyl-2 oxazoline) at the air-water interface studied at the single molecule level. AB - The two-dimensional (2D) center-of-mass diffusion, D, of end-tethered poly(2 methyl-2-oxazoline) (PMOx) lipopolymer chains was studied in a Langmuir monolayer at the air-water interface using wide-field single molecule fluorescence microscopy. In this case, tethering and stabilization of hydrophilic PMOx chains at the air-water interface is accomplished via end-tethering to lipid molecules forming a hydrophobic anchor. To explore the influence of molecular weight, M n, and surface concentration, c s, on lateral mobility, two different PMOx chain lengths of n = 30 and 50 ( n, number of monomer units) were analyzed over a wide range of c s. Using multiparticle tracking analysis of TRITC-labeled PMOx lipopolymers, we found two regimes of lipopolymer lateral mobility. At low c s, D is independent of surface concentration but increases with decreasing n. Here diffusion properties are well described by the Rouse model. In contrast, at more elevated c s, the data do not follow Rouse scaling but are in good agreement with a free area-area model of diffusion. The current study provides for the first time experimental insight into the 2D center-of-mass diffusion of end-tethered polymers at the air-water interface. The obtained results will be of importance for the understanding of diffusion processes in polymer-tethered phospholipid bilayers mimicking biomembranes at low and high tethering concentrations. PMID- 18393537 TI - Colored semitransparent conductive coatings consisting of monodisperse metallic single-walled carbon nanotubes. AB - Single-walled carbon nanotubes (SWNTs) are promising materials for transparent conduction as a result of their exceptional electrical, optical, mechanical, and chemical properties. However, since current synthetic methods yield polydisperse mixtures of SWNTs, the performance of SWNT transparent conductive films has previously been hindered by semiconducting species. Here, we describe the performance of transparent conductors produced using predominantly metallic SWNTs. Compared with unsorted material, films enriched in metallic SWNTs can enhance conductivity by factors of over 5.6 in the visible and 10 in the infrared. Moreover, by using monodisperse metallic SWNTs sorted with angstrom level resolution in diameter, semitransparent conductive coatings with tunable optical transmittance can be produced. PMID- 18393538 TI - Electroluminescence from nanocrystals in an electromigrated gap composed of two different metals. AB - We present a technique for making nanoscale gaps with work function offsets based on electromigrating leads composed of two different metals. Electroluminescence spectra from plain metal gaps with and without CdSe/ZnS (core/shell) nanocrystals are qualitatively very similar and exhibit features that are much broader than the photoluminescence spectra obtained from the same nanocrystals. These observations can be explained by inelastic scattering of conduction electrons in the metal leads or by electroluminescence from small metallic clusters that can form during the fabrication process. However, electroluminescence that spectrally coincides with nanocrystal photoluminescence can be observed in devices containing nanocrystals formed by electromigrating Pt leads bridged with small indium islands. This suggests that electromigrating leads made of different metals is a promising route to fabricating nanoscale gaps with work function offsets for optoelectronic devices. PMID- 18393539 TI - Feasibility of analyzing fine particulate matter in air using solid-phase extraction membranes and dynamic subcritical water extraction. AB - We have evaluated the feasibility of using Empore solid-phase extraction (SPE) membranes as an alternative to conventional techniques for sampling fine airborne particulate matter (PM), including nanoparticles, utilizing a scanning mobility particle sizer (SMPS) and a condensation particle counter to evaluate their efficiency for trapping fine particles in the 10-800 nm size range. The results demonstrate that the membranes can efficiently trap these particles and can then be conveniently packed into an extraction cell and extracted under matrix solid phase dispersion (MSPD) conditions. The potential utility of sampling PM using Empore membranes followed by dynamic subcritical water extraction (DSWE) for fast, efficient, class-selective extraction of polycyclic aromatic hydrocarbons (PAHs) associated with the particles, prior to changing the solvent and analysis by GC/MS, was then explored. The performance of the method was tested using National Institute of Standards and Technology (NIST)-certified "urban dust" reference material (SRM 1649a) and real samples collected at a site in central Rome with heavy road traffic. The method appears to provide comparable extraction efficiency to that of conventional techniques and with using GC/MS, detection limits ranged in the few picograms per cubic meter level. Sampling PM by Empore membranes may reduce the risks of losses of semivolatile compounds, while allowing relatively high sampling flow rates and safe sample storage. Moreover, the combination of MSPD with DSWE permits specific fractions of the PM components to be eluted, thereby generating clean extracts and reducing both analysis time and sample manipulation. PMID- 18393540 TI - Thermogravimetry coupled to single photon ionization quadrupole mass spectrometry: a tool to investigate the chemical signature of thermal decomposition of polymeric materials. AB - Mass spectrometry (MS) is an established analytical technique to analyze evolved gas in thermogravimetry (TG). In this study, for the first time a novel SPI-MS technique using an electron beam pumped VUV excimer lamp as photon source (lambda = 126 nm) was employed in conjunction with thermogravimetry. The coupling was achieved with an improved heated interface and adjacent transfer capillary between TG and ion source of a quadrupole mass spectrometer. The feasibility of this approach was proven by investigating semivolatile substances such as long chain alkanes (heptadecane C17H36), polymers, e.g., polystyrene, polycarbonate, and acrylonitrile-butadiene-styrene, polymer mixtures and blends. Mass spectra with almost no fragmentation were obtained, and quantification of selected substances could be achieved. Polymer mixtures could be distinguished by their SPI mass spectra, and the effect of premixing of polymers has been accessed. Its unique attributes render the TA-SPI-MS method a promising new tool for quantitative and qualitative evaluation of complex organic thermal degradation products. PMID- 18393541 TI - On the nonsingle-site character of Bis(2-dimethylsilyl-indenyl)zirconium(IV) dichloride/MAO and Bis(2-trimethylsilyl-indenyl)zirconium(IV) dichloride/MAO: polymerization characteristics and mechanistic implications. AB - Ethene polymerization with bis(2-dimethylsilyl-indenyl)zirconium(IV) dichloride (1)/MAO and bis(2-trimethylsilyl-indenyl)zirconium(IV) dichloride (2)/MAO and ethene-co-1-hexene polymerization with 1/MAO are presented. The end group analysis of homopolymers reveals a pronounced dependence of the termination rate on temperature changes. In combination with the high molecular weights obtained, these results are in accord with theoretical predictions. Gel permeation chromatography, Fourier transform infrared, and 13C NMR analyses of copolymerization products from 1/MAO as a function of comonomer concentration at two different temperature series denote its tendency to form inhomogeneous polymer blends. Thermal analysis and fractionation results of one such blend indicate an inhomogeneity in the enchainment process and the existence of multiple active sites of differing geometry. These indications are further supported by AMBER force field and density functional theory studies of the catalyst precursors and the active site of 1/MAO. For this system, delta-agostic interactions for the stabilization of the zirconium cation are favored over beta agostic interactions, which, in contrast to the situation in studies on bis-Cp systems, is a sparsely populated species. The gap in activation enthalphies for beta-hydride transfer and elimination is marginalized for these bulky zirconocenes, and conceptually new mechanisms for the isomerization of the vinyl end groups are discussed. Further, unexpected activation of the silicon-hydrogen bond within the ligand framework is observed with an activation enthalpy as low as 14 kcal/mol. PMID- 18393542 TI - Geometries and stabilities of the carbon clusters with the rhodium impurity: a computational investigation. AB - A density functional study of the RhCn(n = 1-6) clusters with different spin states has been carried out systematically by using the B3LYP/Lan2DZ method. The equilibrium geometries associated with total energies and natural populations of RhCn (n = 1-6) clusters are calculated and presented. Stabilities and electronic properties are discussed in detail. The relative stabilities in term of the calculated fragmentation energies show that the lowest-energy RhCn clusters with rhodium atom being located at terminal of carbon chain are the linear geometries and the ground states of the RhCn clusters alternate between doublet (for n-odd members) and quartet (for n-even members) states. Furthermore, the calculated fragmentation energies of the RhCn show strong even-odd alternations: the RhCn clusters with an odd number of carbon atoms are more stable than those with an even number ones. In addition, we comment on the charge transfer and chemical bonding properties within the clusters. PMID- 18393543 TI - Tautomeric and conformational properties of malonamic acid methyl ester, NH2C(O) CH2-C(O)OCH3: electron diffraction and quantum chemical study. AB - The tautomeric and conformational properties of malonamic acid methyl ester, NH2C(O)-CH2-C(O)OCH3, have been investigated by means of gas-phase electron diffraction (GED) and quantum chemical calculations (HF, B3LYP, and MP2 approximations with different basis sets up to 6-311++G(3df,pd)). Both quantum chemistry and GED at 360(8) K result in the existence of a single diketo conformer in the gas phase. According to GED refinement, this conformer possesses an (ac, sc) conformation with dihedral angles C-C-C(NH2)=O of 140.3(3.0) degrees and C-C-C(OCH3)=O of 31.1(7.2) degrees. The experimental geometric parameters are reproduced very closely by MP2 and B3LYP methods with large basis sets. PMID- 18393544 TI - Phase transitions during heating of melt-drawn ultrahigh molecular weight polyethylenes having different molecular characteristics. AB - Changes in the crystalline structure during heating of melt-drawn ultrahigh molecular weight polyethylenes (UHMW-PEs) having different molecular characteristics were analyzed by in situ wide-angle X-ray diffraction measurements. A phase transition from the orthorhombic into the hexagonal phase was observed for all samples, but the perfection was enhanced and the possible temperature window for the hexagonal phase was greater for the sample containing only a higher molecular weight component. In contrast, an increase in retractive stress during heating was confirmed for the sample containing a lower molecular weight component, reflecting melting of the folded-chain crystal (FCC). Differential scanning calorimetry and transmission electron microscopy revealed the dependency of the molecular characteristics of the sample on the resultant morphologies. These results demonstrate that the existence of FCC determines both the quality and the width of the temperature window for the hexagonal phase during heating of melt-drawn UHMW-PEs. PMID- 18393545 TI - Low-radii transitions in co-assembled cationic-anionic cylindrical aggregates. AB - We investigate the formation of charged patterns on the surface of cylindrical micelles from co-assembled cationic and anionic amphiphiles. The competition between the net incompatibility chi (which arises from the different chemical nature of oppositely charged molecules) and electrostatic interactions (which prevent macroscopic segregation) results in the formation of surface domains. We employ Monte Carlo simulations to study the domains at thermal equilibrium. Our results extend previous work by studying the effect of the Bjerrum length l(B) at different values of the cylinder's radius R and chi and analyze how it affects the transition between helical, ring, and isotropic patterns. A critical surface in the space (l(B), R, chi) separating these three phases is found, and we show how it corresponds to a first-order phase transition. This confirms that the Bjerrum length l(B) is a significant parameter in the control of the helical-ring transition; the ring pattern is strongly associated with short-range forces, whereas the helical pattern develops from dominant long-range electrostatic interactions. PMID- 18393546 TI - Physical, chemical, and chemical-physical double network of zwitterionic hydrogels. AB - Zwitterionic hydrogels are very promising for biomedical applications. They are usually copolymerized with other polymers to improve their mechanical properties often at the expense of their biological properties. In this study, physically cross-linked poly(sulfobetaine methacrylate) (polySBMA) hydrogels were prepared, and their physical properties including phase behavior were investigated. Linear polySBMAs, with an average molecular weight ranging from 20.9 kDa to 316 kDa, were prepared via free radical polymerization at different KCl concentrations. The opaque-transparent phase transition of polySBMA-water mixtures were measured using a UV-vis spectrometer. Analysis from dynamic rheometry showed the formation of physically cross-linked hydrogels with mechanical ductility due to reversible charge interactions. Chemically cross-linked hydrogels were also prepared, and their swelling and mechanical properties were evaluated. It was found that the introduction of cross-linkers could lead to a decrease in the amount of physical cross-links in chemical hydrogels. In order to improve the mechanical properties of SBMA hydrogels, linear polySBMA was introduced to the network of chemically cross-linked polySBMA gels, creating a chemical-physical double network (DN) with both chemical and physical cross-links. The chemical-physical DN provides a desirable method to improve the mechanical properties of zwitterionic hydrogels without introducing other hydrophobic moieties. PMID- 18393547 TI - Atomistic insights into the inhibition of cysteine proteases: first QM/MM calculations clarifying the regiospecificity and the inhibition potency of epoxide- and aziridine-based inhibitors. AB - Epoxides and aziridines are important building blocks for inhibitors of cysteine proteases which are promising drug targets for many diseases. In spite of the large amount of experimental data concerning inhibition potency, structure activity relationships, and structural arrangements of enzyme-inhibitor complexes, little is known about the basic principles which connect the substitution pattern with the resulting activities. To shed some light on this issue which is essential for the rational design of improved compounds, we have studied the inhibition processes theoretically for various inhibitors using quantum mechanical/molecular mechanical hybrid approaches and classical molecular dynamics simulations. The careful analysis of the computational results allows insight into the interactions which govern the regio- and stereospecificity of the interactions. Known structure-activity relationships are rationalized in terms of the same interactions that determine the measured pH dependencies. Inconsistencies in existing X-ray structures are resolved through comparison with the computed structures, which leads to a reassessment of the factors that control the inhibition potency. Similarities and differences in the mode of action of epoxide- and aziridine-based inhibitors are elucidated. Finally the small reaction barriers computed for the irreversible step in E64 analogues call into question the commonly accepted two-step model of inhibition since the second, irreversible step is predicted to be so fast that suitably oriented enzyme-inhibitor complexes will react rather than dissociate and equilibrate. PMID- 18393548 TI - Influence of external factors on the micellization process and aggregate structure of poly(oxy)styrene-poly(oxy)ethylene block copolymers. AB - Solutions of the polyoxystyrene-polyoxyethylene block copolymer SO(17)EO(65), where SO denotes polystyrene oxide block as the hydrophobic block and EO the polyethylene oxide block as the hydrophilic block, in mixtures of water (a selective solvent for the EO blocks) and 1,4-dioxane (a good solvent for both blocks) were studied by surface tension and light scattering measurements. Surface and micellar structural parameters have been analyzed as a function of solvent composition. The critical micelle concentration increases and the micellar aggregation number decreases, respectively, as the amount of 1,4-dioxane in the binary solvent increases as a consequence of the enhanced solubility of the SO blocks in the solvent mixture, causing the lowering of the interfacial tension between the hydrophobic blocks in the micellar core and the solvent; therefore, to achieve thermodynamic equilibrium, the micelle size decreases. In addition, static light scattering (SLS) has been proved to be a useful technique to detect the lower boundary of the transition between a dilute micellar solution (sol) to a local-ordered micellar solution (soft gel) resulting from a percolation mechanism. Comparison of the sol-soft gel boundaries obtained from SLS for copolymers SO(17)EO(65) and EO(67)SO(15)EO(67) with those previously derived by rheology is made. Finally, changes in the autocorrelation function of the solutions at the boundary obtained from dynamic light scattering are also analyzed. PMID- 18393550 TI - Spatial arrangement of alpha-cyclodextrins in a rotaxane. Insights from free energy calculations. AB - The rotaxane formed by alpha-cyclodextrins (alpha-CDs) threaded onto a poly(ethylene glycol) (PEG) chain was investigated in the gas phase and in an aqueous solution by means of molecular dynamics simulations. The free-energy difference between the three possible spatial arrangements of consecutive alpha CD--viz.. head-to-head (HH), head-to-tail (HT), and tail-to-tail, was determined using free-energy perturbation calculations. These simulations reveal that the interaction of alpha-CD with the PEG chain is very similar in the two surroundings, whereas the mutual interaction of the macrocycles is stronger in the gas phase than in the aqueous solution. Moreover, difference in the overall interaction appears to stem primarily from changes in the electrostatic contribution. Analysis of intermolecular hydrogen bonds indicates that hydrogen bonds created between alpha-CD and water molecules weaken the hydrogen-bonding interaction of adjacent alpha-CDs. Comparison of the free-energy differences characteristic of the three possible spatial arrangements of contiguous alpha-CDs reveals that the HH motif of the rotaxane is the most stable in the gas phase due to the hydrogen-bond formation between the secondary hydroxyl groups of the two alpha-CDs, and the slight preference for the HT motif in aqueous solution, which can be related to the directionality of the dipole moment borne by the macrocycles in these two motifs. PMID- 18393549 TI - Pulsed electron nuclear double resonance studies of carotenoid oxidation in Cu(II)-substituted MCM-41 molecular sieves. AB - Carotenoid (Car) radical intermediates formed upon catalytic or photooxidation of lutein (I), 7'-apo-7',7'-dicyano-beta-carotene (II), and lycopene (III) inside Cu(II)-MCM-41 molecular sieves were studied by pulsed electron nuclear double resonance (ENDOR) spectroscopies. The Davies and Mims ENDOR spectra (15-20 K) were simulated using the hyperfine coupling constants predicted by density functional theory (DFT) calculations. The DFT calculations revealed that upon chemical oxidation, carotenoid radical cations (Car*+) are formed, whereas carotenoid neutral radicals (#Car*) are produced by proton loss (indicated by #) from the radical cation. This loss is to first order independent of polarity or hydrogen bonding for carotenoids I, II, or III inside Cu(II)-MCM-41 molecular sieves. PMID- 18393551 TI - Properties of free surface of water-methanol mixtures. Analysis of the truly interfacial molecular layer in computer simulation. AB - Molecular dynamics simulations of the vapor-liquid interface of water-methanol mixtures of five different compositions were performed on the canonical (N,V,T) ensemble at 298 K. In addition, the vapor-liquid interface of the two neat systems was simulated, as well. The obtained configurations were analyzed by means of the novel identification of truly interfacial molecules method, which provides a full list of the molecules that are right at the surface (i.e., at the boundary of the two phases). The molecular level roughness of the surface, the adsorption of the methanol molecules at the surface layer, the orientation of the surface molecules, the residence time of the molecules at the surface layer, as well as the surface aggregation of the molecules were analyzed in detail. Both the frequency and the amplitude of the surface roughness were found to become larger with an increasing methanol content. This effect was found to be stronger for the amplitude, which falls in the range of 2-4 A, depending on the composition of the system. Methanol was found to be adsorbed at the surface layer, being preferentially at the humps of the molecularly rough surface. Surface methanol prefers to orient in such a way that the O-CH(3) bond remains perpendicular to the macroscopic plane of the surface, pointing the methyl group to the vapor phase. The main orientational preference of the water molecules is to lie parallel to the surface. Methanol was found to remain considerably longer at the surface layer of the mixed systems than water. Thus, contrary to the fact that the residence times of the two molecules were found to be rather similar to each other at the surface of their neat liquids, the residence time of the methanol molecules was an order of magnitude larger than that of water molecules at the surface of their mixtures. A strong lateral microscopic segregation of the molecules was observed at the surface layer; the minor component of the system (irrespective of whether it was water or methanol) was found to form two dimensional aggregates, leaving the rest of the surface empty for the major component. The effect of the vicinity of the vapor phase on the properties of the molecules was found to vanish very quickly: the composition of the second layer as well as the properties of the molecules of this layer (e.g., dynamics and orientation) did not differ considerably from those in the bulk liquid phase. PMID- 18393552 TI - Blends of polybenzimidazole and poly(vinylidene fluoride) for use in a fuel cell. AB - We report a new blend system consisting of an amorphous polymer polybenzimidazole (PBI) and a semicrystalline polymer poly(vinylidene fluoride) (PVDF). A systematic investigation of the blend pair in various compositions using Fourier transform infrared (FT-IR) spectroscopy provides direct evidence of specific hydrogen bonding interaction involving the N-H groups of PBI and the >CF(2) groups of PVDF. Blending shows a maximum 30 cm(-1) frequency shift in the N-H stretching band of PBI and also the existence of a partial double bond character in the PVDF chain. Differential scanning calorimetry (DSC) study proves the miscibility of these polymers in a wider composition range. The decrease of the T(g) with increasing PVDF in the blend and also the decrease of both the T(m) and T(c) with increasing PBI in the blend attribute the miscibility of the blend systems. The PA doping level of the blend membranes improves significantly as a result of the hydrophobic nature of the PVDF component. PMID- 18393553 TI - Raman study of the rigidity of penta-p-phenylene derivatives used as legs in molecular tripods. AB - Molecular planarity of penta-p-phenylene (P5P) and several substituted derivatives with four side chains of various lengths, including deca(ethylene glycol) groups, is discussed by considering the changes in the intensity ratio between the Raman bands recorded at 1280 and 1220 cm(-1). The intensity ratio between both bands I(1280)/I(1220) shows a small increase with the size of the substituent, indicating a high rigidity for all these compounds, even those with long oligo(ethylene glycol) side chains. This result is important given that these phenylene derivatives are versatile building blocks for the construction of nanometric tripod-shaped adsorbates for biological applications since the side chains should prevent the nonspecific interaction with proteins. PMID- 18393554 TI - Nanoaggregates of copper porphyrazine in floating layers and Langmuir-Schaefer films. AB - Aggregation behavior of unsubstituted copper porphyrazine (CuPaz) on the water surface was studied by analysis of compression curves, Brewster angle microscopy (BAM), and optical spectroscopy. The structure and stability of the CuPaz aqua aggregates in the floating layers are determined by hydration degree that depends on initial surface concentration and surface pressure. Langmuir-Schaefer (LS) films of CuPaz were prepared by deposition of the variously structured floating layers and studied by X-ray scattering technique and optical spectroscopy. Stable and labile structures were detected and compared with the floating CuPaz aqua aggregates. Conditions of formation of the stable four-stacked nanoaggregates in LS films were determined. A model comprising both nucleation of CuPaz on the water surface and structural transformations in the solid films is proposed. PMID- 18393555 TI - Photocurrent generation in noncovalently assembled multilayered thin films. AB - Multilayered photocurrent generating thin films were fabricated by templated noncovalent assembly via stepwise assembly of molecular components. Each of films I-IV contained an underlying self-assembled monolayer (SAM) consisting of an alkanethiol linked covalently to a 2,6-dicarboxypyridine ligand that served as a binding site for attaching additional molecular components. The SAM subsequently was functionalized by sequential deposition of Cu(II), Co(II), or Fe(III) ions followed by a variety of substituted 2,6-dicarboxypyridine ligands as a means to incorporate one or more layers of pyrene chromophores into the film. The films were characterized by contact angle measurements, ellipsometry, grazing incidence IR, cyclic voltammetry, and impedance spectroscopy after deposition of each layer, confirming the formation of ordered, stable layers. Following incorporation into a three-electrode system, photoexcitation resulted in the generation of a cathodic photocurrent in the presence of methyl viologen and an anodic photocurrent in the presence of triethanolamine. Using this strategy, systems were fabricated that produced up to 89 nA/cm(2) of reproducible photocurrent. PMID- 18393556 TI - Thermoactuated diffusion control in soft matter nanofluidic devices. AB - The diffusive transport rate in a soft matter nanofluidic device is controlled with a thermoactuated hydrogel valve. The device consists of three giant unilamellar vesicles linearly conjugated by lipid nanotubes, with a solution of the stimuli-responsive polymer poly(N-isopropyl acrylamide) (PNIPAAm) in the central vesicle. The valve states "high (transport) rate" and "low (transport) rate" are obtained by heat-activated switching between PNIPAAm's dissolved and compact aggregated states. We show that three parameters influence the diffusion rate within the device: the increase of the transport rate caused by a decrease in PNIPAAm concentration upon compaction, the temperature dependence of the buffer viscosity, and the volume excluded by the PNIPAAm hydrogel compartment. PMID- 18393557 TI - Adsorption of cationic starch on cellulose studied by QCM-D. AB - The adsorption of cationic starch (CS) from aqueous electrolyte solutions onto model cellulose film has been investigated by the quartz crystal microbalance with dissipation monitoring (QCM-D) and X-ray photoelectron spectroscopy (XPS). The influence of the electrolyte composition and charge density of CS was examined. The adsorption of CS onto cellulose followed the general trends expected for polyelectrolyte adsorption on oppositely charged surfaces, with some exceptions. Thus, as result of the very low surface charge density of the cellulose surface, highly charged CS did not adsorb in a flat conformation even at low ionic strength. The porosity of the film, however, enabled the penetration of coiled CS molecules into the film at high electrolyte concentrations. Differences between the adsorption behavior of CS on cellulose and earlier observations of the adsorption of the same starches on silica could be explained by the different morphologies and acidities of the hydroxyl groups on the two surfaces. PMID- 18393558 TI - Self-assembled monolayers containing terminal mono-, bis-, and tris nitrilotriacetic acid groups: characterization and application. AB - We have undertaken a structural and functional study of self-assembled monolayers (SAMs) formed on gold from a series of alkylthiol compounds containing terminal multivalent chelators (MCHs) composed of mono-, bis-, and tris-nitrilotriacetic acid (NTA) moieties. SAMs were formed from single-component solutions of the mono , bis-, and tris-NTA compounds, as well as from mixtures with a tri(ethylene glycol)-terminated alkylthiol (EG(3)). Contact angle goniometry, null ellipsometry, and infrared spectroscopy were used to explore the structural characteristics of the MCH SAMs. Ellipsometric measurements show that the amount of the MCH groups on surfaces increases with increasing mol % of the MCH thiols in the loading solution up to about 80 mol %. We also conclude that mixed SAMs, prepared in the solution composition regime 0-30 mol % of the MCH thiols, consist of a densely packed alkyl layer, an amorphous ethylene glycol layer, and an outermost layer of MCH groups exposed toward the ambient. Above 30 mol %, a significant degree of disorder is observed in the SAMs. Finally, functional evaluation of the three MCH SAMs prepared at 0-30 mol% reveals a consistent increase in binding strength with increasing multivalency. The tris-NTA SAM, in particular, is enabled for stable and functional immobilization of a His6-tagged extracellular receptor subunit, even at low chelator surface concentrations, which makes it suitable for applications when a low surface density of capturing sites is desirable, e.g., in kinetic analyses. PMID- 18393559 TI - Effects of self-generated wind on compensational recovery of escape direction in unilaterally cercus-ablated crickets, Gryllus bimaculatus. AB - The effects of self-generated wind on the compensational recovery of escape direction were investigated in unilaterally cercus-ablated crickets, Gryllus bimaculatus. To separate walking and self-generated wind during walking, unilaterally cercus-ablated crickets were placed on a styrofoam ball that was easily rotated by leg motion during walking. The stationary walking on the ball did not produce self-generated wind, because no body motion occurred. Crickets that were trained on the ball but given no artificial air puff for 14 days after cercal ablation did not show any compensational recovery of escape direction. Therefore, spontaneous walking itself was not sufficient to compensate the wind evoked escape direction in the crickets. Artificial air puffs from the anterior direction synchronized with the stationary walking were effective for the compensational recovery of escape direction, but those from the posterior direction were not. As most of the spontaneous walking was directed to the forward direction, only an artificial air puff from the anterior direction coincided well with actual self-generated wind occurring during the onset of normal walking. Therefore, self-generated wind during walking seems essential for the compensational recovery of escape direction in unilaterally cercus-ablated crickets. When artificial air puffs were unsynchronized with walking, no compensational recovery was observed. This result suggests that artificial air puffs should be given just after the onset of spontaneous walking. Otherwise, the artificial air puffs may not be recognized as self-generated wind. PMID- 18393560 TI - Effects of mechanical stretching on caspase and IGF-1 expression during the proliferation process of myoblasts. AB - It has been reported that the synthesis, degradation, and metabolism of muscle proteins in myoblasts, as well as the proliferation and differentiation of cells, are influenced by various related to extracellular signaling molecules, such as neural transmitters, growth factors, and hormones, when muscle tissue has been exposed to mechanical stimulation. However, reports regarding the expression of growth factors during mechanical stimulation of myoblasts are few, and many questions remain unanswered. We examined the mRNA expression of insulin-like growth factor 1 (IGF-1) in myoblasts subjected to mechanical stretching in vitro. In addition, apoptosis caused by intracellular stress has been reported to occur during muscle development at the embryonic stage. To clarify the expression of intracellular stress factors, we here investigated related gene expression. Expression of IGF-1 increased in the early stage of cell stretching, followed by a decrease in the late stage. This suggests that mechanical stimulation resulted in an immediate increase in IGF-1 expression, followed by a decrease as cells acclimated to the inducing environment. Caspase was significantly expressed in a stretch group at 12 hours after the beginning of mechanical stimulation, compared with a control group. This suggests that cellular proliferation is also regulated by intracellular stress factors involving the endoplasmic reticulum, mitochondria, and other organelles during the process of muscle proliferation and differentiation. PMID- 18393561 TI - Maternal inheritance of mitochondrial DNA (mtDNA) in the Pacific oyster (Crassostrea gigas): a preliminary study using mtDNA sequence analysis with evidence of random distribution of MitoTracker-stained sperm mitochondria in fertilized eggs. AB - In many bivalve species, paternal and maternal mitochondrial DNA (mtDNA) from sperm and eggs is transmitted to the offspring. This phenomenon is known as doubly uniparental inheritance (DUI). In these species, sperm mtDNA (M type) is inherited by the male gonad of the offspring. Egg mtDNA (F type) is inherited by both male and female somatic cells and female gonadal cells. In Mytilidae, sperm mitochondria are distributed in the cytoplasm of differentiating male germ cells because they are transmitted to the male gonad. In the present study, we investigated maternal inheritance of mtDNA in the Pacific oyster, Crassostrea gigas. Sequence analysis of two mitochondrial non-coding regions revealed an identical sequence pattern in the gametes and adductor muscle samples taken from six males and five females. To observe whether sperm mitochondria were specifically located in the cytoplasm of differentiating germ cells, their distribution was recorded in C. gigas fertilized eggs by vital staining with MitoTracker Green. Although the 1D blastomere was identified in the cytoplasm of differentiating germ cells, sperm mitochondria were located at the 1D blastomere in only 32% of eggs during the 8-cell stage. Thus, in C. gigas, sperm mitochondria do not specifically locate in the germ cell region at the 1D blastomere. We suggest that the distribution of sperm mitochondria is not associated with germ cell formation in C. gigas. Furthermore, as evidenced by the mtDNA sequences of two non-coding regions, we conclude that mitochondrial DNA is maternally inherited in this species. PMID- 18393562 TI - Origin of the transition from aquatic to terrestrial habits in Nothopsyche caddisflies (Trichoptera: Limnephilidae) based on molecular phylogeny. AB - The larvae and pupae of most caddisflies (Trichoptera) are aquatic, whereas the adults are terrestrial. However, the trichopteran genus Nothopsyche includes species with terrestrial pre-pupal and pupal stages and with an entirely terrestrial life cycle. The phylogenetic relationships within this genus, inferred from sequences of mitochondrial 16S and nuclear 18S ribosomal RNA genes, suggest that Nothopsyche species were originally aquatic and that a single lineage acquired tolerance to terrestrial habitats at the pre-pupal and pupal stages. In this lineage, N. montivaga became completely terrestrial. In addition, the larval case materials changed from plant matter to sand at one point in the phylogeny of this genus. PMID- 18393563 TI - Genetic differentiation of the Fejervarya limnocharis complex from Bangladesh and other Asian countries elucidated by allozyme analyses. AB - The present study was conducted to elucidate the genetic divergence and the phylogenetic relationships in the F. limnocharis complex from Bangladesh and other Asian countries such as Sri Lanka, Thailand, Malaysia, Taiwan and Japan by allozyme analyses. We used a total of 95 frogs of the F. limnocharis complex from these countries and F. cancrivora from the Philippines as an outgroup. Based on body size, the F. limnocharis complex from Bangladesh was divided into three distinct groups: large, medium and small types. Allozyme analyses were carried out with 28 loci encoding 20 enzymes and two blood proteins by horizontal starch gel electrophoresis. When genetic distance was calculated, distinct divergence was found among the three types: mean genetic distance was 0.782 between the small and medium types, 1.458 between the large and medium types, and 1.520 between the large and small types. Phylogenetic trees based on genetic distance showed that all populations of Bangladesh small type strongly formed a cluster and were found to be most closely related to the Sri Lanka population; that all populations of Bangladesh large type formed a very strong cluster and were grouped with several populations from Thailand, Malaysia, Japan, and Taiwan; and that the medium type was segregated from all other groups. This may imply that each of the three types is a different species, and that the medium type is possibly an undescribed taxon. PMID- 18393564 TI - A trend of central versus peripheral structuring in mitochondrial and nuclear gene sequences of the Japanese wood mouse, Apodemus speciosus. AB - A phylogeographic analysis was performed on Japanese endemic wood mice (Apodemus speciosus) using nuclear interphotoreceptor retinol binding protein (IRBP) gene sequences (1,152 bp), together with previously published mitochondrial cytochrome b (cyt b) data. In the IRBP analysis, 40 haplotypes were recovered from 84 individuals by statistical and subcloning methods. Substantial sequence variation was determined from the IRBP data (pi=0.0047), and no significant evidence of recombination was detected. From the phylogenetic analysis, the 40 haplotypes fell into two major groups with geographic associations, irrespective of the karyotype groups (2n=46 and 2n=48), yielding a trend of central (Hokkaido, Honshu, Shikoku, Kyushu, and Sado) and peripheral (Izu, Oki, Tsushima, and Satsunan Is.) groupings. This geographic pattern is similar to that observed in the cyt b data, with a different insular grouping of Sado, Hokkaido, Izu, and Satsunan islands, and also to that of morphological features. In both gene data sets, nested clade analyses revealed allopatric fragmentation in the "peripheral island clades" and range expansion in the "central island clades." A mismatch analysis using cyt b data also suggested expansion of the central islands clade. Thus, the trend of central vs. peripheral structuring may be attributable to past demographic dynamics in the two distinct haplotype clades, such as range expansion of one clade in the central area of the Japanese Islands, leaving the other clade in the periphery. PMID- 18393565 TI - Experimental test of intraspecific competition mechanisms among tadpoles of Leptodactylus ocellatus (Anura: Leptodactylidae). AB - Intraspecific competition is predicted to strongly influence species abundance and dynamics through two main mechanisms: consumption and interference of resources. Tadpoles were used in experiments in which we tried to elucidate the relative importance of each mechanism. Our goal was to apply this experimental procedure to Leptodactylus ocellatus, a common South American anuran, a species whose larvae exhibit aggregative behavior and receive parental care. Previous work suggests that tadpole schools should present lower levels of intraspecific competition. Tadpoles from a single nest were reared in the laboratory in three densities (1, 2, and 4 individuals/container) and three food levels (1, 2, and 4 ration multiples) in a randomized three-block design for a factorial analysis of variance, up to day eight. Contrary to previous work with other species, our results show both the absence of interference competition effects, and that larval growth depends only on per capita food availability. The differences between species in intraspecific competition mechanisms are probably related to strong differences in ecology and life history. Leptodactylus ocellatus tadpoles could be directing interference competition away from their kin, reducing schooling costs. Further studies (including kinship as a factor) would give more information about these larvae, allowing a better understanding of the evolutionary and ecological mechanisms behind the biological patterns observed in Leptodactylus species. PMID- 18393566 TI - Corticosterone responses to capture and restraint in emperor and Adelie penguins in Antarctica. AB - Birds respond to capture, handling and restraint with increased secretion of corticosterone, a glucocorticoid hormone that helps birds adjust to stressful situations. Hoods are reported to calm birds, but possible effects of hoods on corticosterone responses have not been reported for any bird. Corticosterone responses to restraint in Adelie penguins held by their legs with their head covered by a hood were markedly lower than responses of penguins restrained in a mesh bag inside a cardboard box (corticosterone at 30 min 15.69+/-1.72 cf. 28.32+/-2.75 ng/ml). The birds restrained by the two methods were sampled at the same location but in different years, so the differences in corticosterone responses cannot unequivocally be ascribed to an effect of hoods to reduce corticosterone responses. Corticosterone responses have been measured in some penguins, but not in the largest, the emperor penguin (Aptenodytes forsteri). The relationship between body mass and corticosterone responses to capture and restraint in penguins was examined in emperor penguins captured on sea ice in McMurdo Sound and Adelie penguins (Pygoscelis adeliae) captured at Cape Bird, Ross Island, Antarctica. Total integrated corticosterone responses were higher in the emperor than the Adelie penguins, but corrected integrated corticosterone responses, which represent the increase in corticosterone from initial concentrations and hence the corticosterone response to restraint, were the same. The results for the emperor and Adelie penguins, together with data from other penguin species, suggest that there is no relationship between the size of corticosterone responses and body mass in penguins. PMID- 18393567 TI - The novel mutant scl of the medaka fish, Oryzias latipes, shows no secondary sex characters. AB - A new mutant that has neither male nor female secondary sex characters was found in the medaka, Oryzias latipes. Both XX and XY mature mutants had gonads with many spermatozoa, but spawning did not occur when the mutants were paired with normal males or normal females. F1 progeny were successfully obtained by artificial insemination using unfertilized eggs from wild-type females and spermatozoa of the XY mutant. The mutant phenotype did not occur in the F1 progeny from this cross. Incrossing among the F1 progeny produced 17 mutant offspring out of 68 progeny (25%), demonstrating that the mutant phenotype is caused by a single recessive mutation. This mutant was named scl (sex character less). Because papillary processes, a male secondary sex character, were induced in the XY mutants by androgen administration, it seems that the androgen receptor is functioning normally. We found a loss-of-function type mutation in the P450c17 gene of the mutant; this gene encodes a steroidogenic enzyme required for the production of estrogen and androgen. The scl phenotype was completely linked to the mutant genotype of P450c17, strongly suggesting that mutation at the P450c17 locus is responsible for the scl mutant phenotype. PMID- 18393568 TI - Chromosomal aberrations in Japanese grass voles in and around an illegal dumpsite at the Aomori-Iwate prefectural boundary. AB - Bone marrow chromosomes of thirty specimens of the Japanese grass vole, Microtus montebelli (2n=30), which had been caught on and near an illegal dumpsite at the Aomori-Iwate prefectural boundary, were analyzed and compared with those of fifteen grass voles from non-polluted areas as part of an effort to assess genotoxic influences on grass voles in the dumpsite area. Fifty metaphases per specimen were examined with particular attention to numerical and structural aberrations. Two specimens from the dumpsite had 2n=31, which was confirmed by G banding analysis to have been caused by centric fission of M6 homologs, while control specimens had no such abnormality. In specimens from the polluted area, the mean number of chromosomal aberrations (breaks and/or gaps) per 50 metaphases per specimen was 2.57+/-0.41, which was significantly higher than that (0.80+/ 0.14; P<0.01) in control specimens. Chromosomal aberrations were randomly distributed on chromosomes, with frequencies being proportional to the relative lengths of chromosomes. Our findings suggest that grass voles at and around the dumpsite have been seriously damaged at the chromosomal level and, moreover, that M. montebelli might be useful as an indicator species for genotoxic assessment of below-ground pollution by industrial waste at illegal dumpsites. PMID- 18393569 TI - Mammalian Sox15 gene: promoter analysis and implications for placental evolution. AB - Sox15 belongs to the Sox (Sry-type HMG box) protein family, which is involved in placental development and muscle regeneration. Previously, we showed that the Sox15 gene is highly expressed in the trophoblast giant cells of the mouse placenta. To elucidate the molecular mechanisms of the tissue-dependent transcription of the gene, we isolated approximately 2.2 kb of the 5'-flanking sequence upstream of the transcription initiation site and used it to construct luciferase reporter plasmids. A variety of cell lines, including trophoblast stem (TS) cells, placenta-derived Rcho-1 cells, and myoblast C2C12 cells, required the same 5'-flanking sequence, from -109 to -8, for basal promoter activity. In contrast, the sequences from -297 to -149 and from -148 to -110 were required for cell-type-specific promoter activity in myoblast-derived C2C12 cells and placenta derived Rcho-1 and TS cells, respectively. These results suggest that the region from -297 to -8 of the Sox15 gene contains three distinct cis-elements that respectively control placenta-specific, myoblast-specific, and common basal expression. We also searched for Sox15 ortholog(s) in the genome databases of various vertebrate species. The results indicated that the three regulatory promoter sequences of the Sox15 genes were conserved among eutherian mammals during vertebrate evolution. Interestingly, the marsupial opossum gene that is closest to Sox15 appeared to be a pseudogene. These findings indicate that Sox15 may have been involved in placental evolution. PMID- 18393570 TI - Characteristic expression of three heat shock-responsive genes during larval diapause in the bamboo borer Omphisa fuscidentalis. AB - Three heat shock-responsive genes, Ofhsp70, Ofhsc70, and Ofhsp90, were identified in Omphisa fuscidentalis. These genes encode proteins with molecular weights of 70, 72, and 82 kDa, respectively, and were upregulated during heat shock. Ofhsp70 was expressed actively in the pre-diapause period, but its expression was low during diapause. While Ofhsc70 expression was low during pre-diapause and the first half of diapause, its expression increased greatly in the second half before diapause termination. In contrast, Ofhsp90 expression decreased by half during the transition to diapause and conspicuously decreased during late diapause. The results suggest that Ofhsp90 is associated with the maintenance of diapause and Ofhsc70 with the termination of diapause, whereas the expression of Ofhsp70 is unrelated to diapause. The insect molting hormone 20-hydroxyecdysone (20E) and juvenile hormone analog (JHA) caused increases in the mRNA expression of Ofhsc70, but not Ofhsp70 or Ofhsp90, in diapausing larvae. These transcriptional responses to 20E are consistent with the changes in ecdysteroid titer in the hemolymph relative to gene expression during diapause. PMID- 18393571 TI - Effects of pituitary glycoprotein hormones and thyroid hormones on in-vitro vitellogenin incorporation into organ-cultured oocytes in the Japanese eel, Anguilla japonica. AB - The objectives of the present study were to establish a long-term culture system for previtellogenic ovarian fragments of the Japanese eel (Anguilla japonica) and to identify the effects of salmon pituitary glycoprotein fraction (SPG), thyroxine (T4), and 3,5,3'-triiodothyronine (T3) on the uptake of vitellogenin (VTG) by cultured ovarian fragments evidenced by the appearance of yolk globules (YGs) within the oocytes. Yolk globules first appeared in the oocytes incubated in media containing only VTG (VTG-only group) after 9 days, whereas YGs began to accumulate in the oocytes of ovarian fragments cultured in media containing VTG+SPG (SPG group) following only 3 days of incubation. Furthermore, the occurrence of vitellogenic oocytes (%VO) and proportion of YGs within oocytes (%YG area) were significantly higher in follicles cultured in 30 ng/ml SPG throughout the culture period. No such stimulatory effects of T4 on VTG uptake were observed. Incubation of ovarian fragments with VTG and T3 (T3 group; 50 ng/ml) resulted in an increased %VO compared to follicles in the VTG group by day 9 of culture, and from day 10 onwards, both %VO and %YG area became significantly higher in follicles of the T3 group. Interestingly, SPG stimulated VTG incorporation and YG accumulation even in small oocytes (approximately 150 microm), whereas T3 showed these effects only in larger sized oocytes (> 180 microm). These results suggest that both SPG and T3 can accelerate VTG incorporation, but the mechanisms whereby this is achieved may differ between these hormone preparations. PMID- 18393573 TI - Accessible and total lead in low-cost jewelry items. AB - The US Consumer Product Safety Commission has proposed a ban on children's metal jewelry containing more than 0.06% lead by weight. This ban would replace the current interim enforcement standard, which also includes a measurement of accessible lead. Under current guidelines, accessible lead for any component of a jewelry item must not exceed 175 microg. Critics argue that lead in jewelry is inaccessible if items are properly plated. The objective of this study was to determine whether highly leaded jewelry also has high accessible lead content. Sixty-four inexpensive, leaded jewelry items with a wide range of total lead content (all above the 0.06% threshold) were tested first for accessible lead and then for total lead, with analysis by atomic absorption spectrometry. Fifty of the items exceeded the 175-microg maximum allowed for accessible lead. Thirty-one of these items exceeded 1000 microg accessible lead, and 18 items exceeded 3000 microg accessible lead. The finding that a majority of tested items had both accessible and total lead content above current standards provides support for the rationale of simplifying the current standard by basing regulations on total lead content. PMID- 18393572 TI - Stability in aromatase immunoreactivity of steroid-producing cells during early development of XX gonads of the Nile tilapia, Oreochromis niloticus: an organ culture study. AB - The organ culture system is a useful tool to study the effects of various factors on the development of undifferentiated gonads. In this study, we first established an organ culture system for gonads of all genetic male and female Nile tilapia at 5-122 days after hatching (dah). This short-term (3 days) organ culture system was then used to examine the stability of the immunoreactivity of aromatase (the enzyme which converts androgen to estrogen, thus playing a crucial role in ovarian differentiation) in steroid-producing cells (SPCs). Immunohistochemical analyses revealed that aromatase-positive cells could be initially detected in the vicinity of blood vessels in the XX gonads at 7 dah. These SPCs completely lost their immunoreactivity after 3 days in culture, indicating the instability of SPCs during early ovarian differentiation. In contrast, the immunoreactivity of the SPCs was maintained to some extent even after 3 days in culture, if the gonads were from 15-23 dah. In XX gonads collected at 122 dah, there were two major populations of SPCs: one in the vicinity of the blood vessel and the other near the oocyte. The aromatase immunoreactivity was maintained in SPCs located around the oocytes, but not in those in the vicinity of the blood vessel, after 3 days in culture. These results suggest that the SPCs originate from the cells in the vicinity of the blood vessels prior to the initial ovarian differentiation in tilapia and that the degree of differentiation of SPCs is dependent on their location in the ovary. PMID- 18393576 TI - Potential environmental contaminant risks to avian species at importnat bird areas in the northeastern United States. AB - Environmental contaminants can have profound effects on birds, acting from the molecular through population levels of biological organization. An analysis of potential contaminant threats was undertaken at 52 Important Bird Areas (IBAs) within the northeastern Atlantic coast drainage. Using geographic information system methodology, data layers describing or integrating contamination (impaired waters, fish or wildlife consumption advisories, toxic release inventory sites, and estimates of pesticide use) were overlaid on buffered IBA boundaries, and the relative threat at each site was ranked. Some species of birds residing at Jefferson National Forrest (NF), Stewart B. McKinney National Wildlife Refuge (NWR), Great Dismal Swamp NWR, Blue Ridge Parkway, Shenandoah National Park (NP), Adirondack Park, Edwin B. Forsythe NWR, George Washington NF, Green Mountain NF, Long Island Piping Plover Beaches, and Merrymeeting Bay may be threatened by environmental contaminants. These sites exhibited moderate to high percentages of impaired waters and had fish consumption advisories related to mercury and polychlorinated biphenyls, and were located in counties with substantial pesticide use. Endangered, threatened, and Watch List bird species are present at these sites. The Contaminant Exposure and Effects-Terrestrial Vertebrates database was searched within buffered IBA boundaries, and for a moderate number of sites there was concordance between the perceived risk and contaminant exposure. Several of the IBAs with apparently substantial contaminant threats had no avian ecotoxicological data (e.g., George Washington NF, Shenandoah NP). Based upon this screening level risk assessment, contaminant biomonitoring of birds is warranted at such sites, and data generated from these efforts could foster natural resource management activities. PMID- 18393577 TI - A "weight of evidence" approach for the integration of environmental "triad" data to assess ecological risk and biological vulnerability. AB - A new Expert Decision Support System (EDSS) that can integrate Triad data for assessing environmental risk and biological vulnerability at contaminated sites has been developed. Starting with ecosystem relevance, the EDSS assigns different weights to the results obtained from Triad disciplines. The following parameters have been employed: 1) chemical soil analyses (revealing the presence of potentially dangerous substances), 2) ecotoxicological bioassays (utilizing classical endpoints such as survival and reproduction rates), 3) biomarkers (showing sublethal pollutant effects), and 4) ecological parameters (assessing changes in community structure and functions). For each Triad discipline, the EDSS compares the data obtained at the studied field sites with reference values and calculates different 0-1 indexes (e.g., Chemical Risk Index, Ecotoxicological Risk Index, and Ecological Risk Index). The EDSS output consists of 3 indexes: 1) Environmental Risk index (EnvRI), quantifying the levels of biological damage at population-community level, 2) Biological Vulnerability Index (BVI), assessing the potential threats to biological equilibriums, and 3) Genotoxicity Index (GTI), screening genotoxicity effects. The EDSS has been applied in the integration of a battery of Triad data obtained during the European Union-funded Life Intervention in the Fraschetta Area (LINFA) project, which has been carried out in order to estimate the potential risk from soils of a highly anthropized area (Alessandria, Italy) mainly impacted by deposition of atmospheric pollutants. Results obtained during 4 seasonal sampling campaigns (2004-2005) show maximum values of EnvRI in sites A and B (characterized by industrial releases) and lower levels in site D (affected by vehicular traffic emissions). All 3 potentially polluted sites have shown high levels of BVI and GTI, suggesting a general change from reference conditions (site C). PMID- 18393578 TI - An assessment of endocrine disruption in mollusks and the potential for developing internationally standardized mollusk life cycle test guidelines. AB - This paper summarizes what is known about the endocrine systems of mollusks and how they can be disrupted by exogenous substances. It then examines the various possibilities for using mollusk-based toxicity tests to detect and assess the environmental risks of endocrine-disrupting chemicals (EDCs). It is concluded that there are no internationally standardized tests with mollusks available at present that are suitable for assessing the risks of long-term exposure to EDCs but that several published methods show potential. At the present state of knowledge, the most promising mollusk species for use in partial life cycle testing is probably the fresh- and brackish-water mesogastropod, Potamopyrgus antipodarum, which is known to be responsive to both direct and indirect androgens and to estrogens (and their mimics). Less experience of full life cycle testing exists with mollusks, but the freshwater pulmonate gastropod Lymnaea stagnalis at present offers the best possibility for a practical procedure. In both cases, there is a need for substantial test optimization and validation before these procedures could form the basis of international guidelines. PMID- 18393579 TI - The moderating effects of task complexity on the relationship between regulatory foci and safety and production performance. AB - Regulatory foci of promotion and prevention have been shown to relate differentially to occupational safety and production. This research proposes that task complexity can help explain the differences reported between these 2 self regulatory processes and safety and productivity performance. Results revealed that promotion is positively related to production and prevention is positively related to safety regardless of task complexity. However, when task complexity is high, promotion negatively relates to safety and prevention negatively relates production. Implications for work motivation theory and research, as well as avenues for future research, are discussed. Practical implications for managerial interventions to optimize both safety and productivity are also presented. PMID- 18393580 TI - The relationship between long-term job strain and morning and evening saliva cortisol secretion among white-collar workers. AB - The objective of this study was to assess long-term job strain impact on morning and evening salivary cortisol secretion. In all 77 white-collar workers (31% women; sample mean age, 42 years at baseline) volunteered to sample morning (immediately after waking up) and evening (10 p.m.) salivary cortisol for 7 consecutive days. By median split on aggregated self-reported isostrain from three consecutive questionnaires distributed in a period of approximately 3.5 years the participants were classified into a high or low long-term isostrain condition. Regardless of strain condition, there was a significant reduction in morning salivary cortisol secretion from the working week to the weekend, whereas evening salivary cortisol secretion showed no significant variation during the week. Although chronic isostrain did not affect the morning saliva cortisol measures, evening cortisol secretion was significantly elevated in the chronic high isostrain group throughout the whole week. The elevated evening cortisol measures associated with chronic high strain are concordant with the findings in other studies on long-term strain. PMID- 18393581 TI - Testing the effort-reward imbalance model among Finnish managers: the role of perceived organizational support. AB - The present study is aimed at examining the combined effects of effort-reward imbalance (ERI), overcommitment (OVC), and perceived organizational support (POS) on turnover intentions and work engagement, among Finnish managers (n = 1,301). Consequently, the study contributes to the research literature by examining how the ERI-outcomes relationship was dependent simultaneously on OVC and POS. The results showed that ERI x OVC x POS interaction was significant only for turnover intentions. The ERI-turnover intentions relationship was strongest under conditions of high OVC and low POS. In addition, the relationship between ERI and decreased work engagement, especially dedication, was strengthened among overcommitted managers, compared to their less committed counterparts. Altogether, the results indicate that interventions aimed at reducing turnover intentions and increasing work engagement by increasing ERI should consider OVC and POS. PMID- 18393582 TI - Sense of coherence as a protective mechanism among targets of workplace bullying. AB - The present study investigated sense of coherence (SOC) as a protective factor among targets of workplace bullying. A hypothesis that strong SOC lessens the relationship between exposure to bullying and symptoms of posttraumatic stress was tested in a cross-sectional sample of 221 self-labeled targets of workplace bullying. The findings showed that SOC offers most protective benefits to targets exposed to low levels of bullying, whereas the benefits of SOC diminish as bullying becomes more severe. The results support previous findings that workplace bullying is a traumatic experience for those exposed to it, regardless of the target's available coping resources. PMID- 18393583 TI - Effects of racial and sexual harassment on work and the psychological well-being of African American women. AB - Research on workplace harassment has typically examined either racial or sexual harassment, without studying both simultaneously. As a result, it remains unknown whether the co-occurrence of racial and sexual harassment or their interactive effects account for unique variance in work and psychological well-being. In this study, hierarchical linear regression analyses were used to explore the influence of racial and sexual harassment on these outcomes among 91 African American women involved in a sexual harassment employment lawsuit. Results indicated that both sexual and racial harassment contributed significantly to the women's occupational and psychological outcomes. Moreover, their interaction was statistically significant when predicting supervisor satisfaction and perceived organizational tolerance of harassment. Using a sample of African American women employed in an organizational setting where harassment was known to have occurred and examining sexual and racial harassment concomitantly makes this study unique. As such, it provides novel insights and an important contribution to an emerging body of research and underscores the importance of assessing multiple forms of harassment when examining organizational stressors, particularly among women of color. PMID- 18393584 TI - Sex and ethnicity as moderators in the sexual harassment phenomenon: a revision and test of Fitzgerald et al. (1994). AB - Fitzgerald, Hulin, and Drasgow (1994) proposed that personal vulnerability characteristics (such as sex and ethnicity) would moderate the effect of sexual harassment on its outcomes. This paper argues that personal vulnerability characteristics instead moderate the effect of organizational sexual harassment climate on sexual harassment because of their role as identity markers within social hierarchies. Using a sample of nearly 8,000 male and female military personnel from four ethnicity groups, the proposition that organizational climate differentially affects sexual harassment frequency across sex and ethnicity was evaluated. Results suggested that sex is an important moderator of these relationships, but that ethnicity is not. Further, sex and ethnicity were not found to moderate the effect of sexual harassment on its outcomes. Potential generalizability of these results to other types of harassment (e.g., racial harassment, bullying), as well as needed future research in this area, is discussed. PMID- 18393586 TI - The crossover of perceived health between spouses. AB - The current study examined the crossover of perceived health between spouses and the mediating roles of self-esteem and undermining in this process. Data were collected from a sample of 2,108 couples from the Russian Longitudinal Monitoring survey. Using structural equation modeling, the authors tested a crossover model that incorporated three mechanisms: bidirectional crossover of perceived health between spouses, common stressors (income), and indirect mediated effects (social undermining). The model showed an acceptable fit to the data and provided support for all three-crossover mechanisms. Furthermore, self-esteem mediated the relationship between economic hardship and perceived health. The authors discuss theoretical aspects of the crossover of perceived health and implications of our findings for the design of preventive interventions to help family members cope more effectively with economic hardship. PMID- 18393587 TI - Work-family characteristics as determinants of sickness absence: a large-scale cohort study of three occupational grades. AB - This study examined the previously unexplored occupational grade-specific relationships of domestic responsibilities, the age of children, and work-family spillover, with registered sickness absence (>3 days' sick leave episodes, a mean follow-up of 17 months; n = 18,366 municipal employees; 76% women). The results showed that negative spillover from work into family life predicted a heightened rate of sickness absence spells among both women and men in all occupational categories (except upper white-collar men), but especially among blue-collar and lower white-collar employees. Furthermore, among all white-collar employees (except upper white-collar men), having young children (<7 years of age) was predictive of an increased absence rate. Bearing the main responsibility for domestic work did not considerably predict sickness absenteeism in any occupational grade. The authors conclude that some specific work-family characteristics play a role in sickness absence, but their impact on sickness absence partly varies according to gender and occupational grade. PMID- 18393588 TI - Summary of the Caribbean subregional workshop on quality in immunohematology, a collaborative effort to improve international blood transfusion services. PMID- 18393589 TI - Technical issues in neonatal transfusions. AB - Neonatal transfusions provide challenges at several steps in the process. Neonates are often transfused with relatively small volumes at slow flow rates from syringes,whereas at other times they require relatively massive transfusions or exchange transfusions. To facilitate these specialized transfusions, blood banks often modify their procedures to provide small volumes of blood components that are sometimes dispensed in syringes or to reconstitute whole blood for exchange transfusions. Hospitals must implement policies and procedures to ensure that the blood components are transfused safely when using these specialized techniques for infants. Nevertheless, some issues remain in many hospitals, such as the difficulty in safely warming blood components for neonatal transfusions and the difficulties in using approved labels for small containers that are sometimes prepared at the bedside. PMID- 18393590 TI - Neonatal red cell transfusions. AB - This review discusses RBC transfusion in the neonatal age group and explores how one institution arrived at current common practice. Special considerations such as CMV infectious risk and GVHD are discussed. PMID- 18393591 TI - Neonatal and infant platelet transfusions. PMID- 18393592 TI - Neonatal plasma transfusions. PMID- 18393593 TI - West African refugee health in rural Australia: complex cultural factors that influence mental health. AB - Health and mental health practitioners in rural and regional Australia are increasingly being presented with the challenge of working cross-culturally. Due to a diversity of cultures, generic approaches are needed that take account of culture without requiring the practitioner to have detailed knowledge of each person's background. However, there are many practitioners from diverse backgrounds now working in Australia at various stages of enculturation into Western practice. The cultural grounding of one practitioner from West Africa is used to highlight potential areas of misunderstanding, and to offer an accessible point of departure for culturally sensitive practice and formal research opportunities. PMID- 18393594 TI - Vaccine safety: genuine concern or a legacy of unfounded skepticism? PMID- 18393595 TI - Ecological aspects in vaccine trials. PMID- 18393597 TI - Vaccine adjuvant formulation: a relatively neglected field that is crucial to vaccine success. AB - This 3-day conference, with a focus on formulation, is part of a series of conferences organized by Meetings Management on different aspects of vaccine adjuvants. The previous Modern Vaccine/Adjuvants Formulation took place in Prague in 2004 and it was interesting to observe what has changed and what since then remains the same. PMID- 18393598 TI - Bangkok International Conference on Avian Influenza 2008. AB - The Bangkok International Conference on Avian Influenza 2008 was organized by the National Center for Genetic Engineering and Biotechnology of Thailand. The Center has been playing an active role in the organization of research projects and knowledge management in preparation for an influenza pandemic since the beginning of the H5N1 avian influenza (AI) outbreak in the region in 2004. The timing of the conference, January 23rd, coincided with the 2004 announcement of the first outbreak in the country, marking the fourth year of widespread AI outbreaks in this region. In this 4-year period, progresses have been made scientifically and politically. Nevertheless, AI is still a major threat to us all. At the opening ceremony of the conference, Yongyuth Yuthavong, the Minister of Science and Technology of Thailand, emphasized that we are all together in the global village in this AI crisis and that we have to share information and materials and work together in the fight against this enemy of mankind. PMID- 18393599 TI - Tetravalent meningococcal vaccination for infants. PMID- 18393600 TI - STEP trial and HIV-1 vaccines inducing T-cell responses. AB - Recombinant, nonreplicating, human adenovirus serotype 5-vectored vaccine, known as MRKAd5, expressing three HIV-1 clade B-derived internal proteins when used in a homologous immunization regimen, did not decrease HIV-1 infection rate nor postinfection virus load in the first Phase IIb proof-of-concept trial. However, the vaccine did not reach the limits of vaccine T-cell induction and its design can be improved both from the point of the HIV-1-derived immunogens and their delivery. Therefore, failure of the first experimental HIV-1 vaccine focusing on induction of T-cell responses cannot be a reason for dismissal of the whole T cell vaccine concept, nor for losing a positive attitude toward systematic HIV-1 vaccine development. PMID- 18393601 TI - HIV vaccine research in Thailand: lessons learned. AB - Thailand is the only country that has conducted two Phase III efficacy trials and ten other Phase I/II studies using various HIV-1 vaccine constructs. This outstanding record is the result of strong efforts and commitment among various partners, including policy makers, regulators, researchers and foreign collaborators. Recently, it has become apparent that hardly any new HIV/AIDS candidate vaccines are in the pipeline reaching the stage of clinical testing, especially a candidate vaccine that is suitable for Thailand and the surrounding region. However, many lessons learned can be utilized or modified for other vaccine trials, particularly vaccines against other infectious diseases prevalent in developing countries. PMID- 18393602 TI - Tenets of protection from progression to AIDS: lessons from the immune responses to HIV-2 infection. AB - In the past 25 years, life survival curves of many countries have been remodeled owing to HIV infection. Both HIV-1 and HIV-2 can cause AIDS, yet patients infected with HIV-2 fare much better clinically and most will never experience detrimental effects of the infection. Despite over two decades of comprehensive research into vaccine development, a prophylactic vaccine is not yet realized. An essential missing link in the innovation of a successful vaccine strategy is the description of a favorable immune response that abolishes virus replication. Lessons learned from studying the role of the immune system in the long-term nonprogression characteristic of HIV-2 infection will offer insight into how a balanced immune response can protect from the destruction of the immune system associated with chronic HIV-1 infection. PMID- 18393603 TI - Protection against filovirus infection: virus-like particle vaccines. AB - Significant progress has been made in vaccine development against infection by Ebola and Marburg viruses, members of the Filoviridae, which cause severe hemorrhagic fevers in humans with no effective treatment and a mortality rate of up to 90%. Several vaccine strategies have been shown to effectively protect immunized animals against filovirus infection. Among these candidate vaccine strategies, virus-like particles represent a promising approach and have been shown to protect small laboratory animals as well as nonhuman primates against lethal challenge by Ebola and/or Marburg viruses. This review briefly summarizes filovirus epidemiology and pathogenesis, and focuses on the discussion of recent advances in filovirus vaccine development and the current understanding of protective immune responses against filovirus infection with an emphasis on the progress and challenge of filovirus virus-like particle vaccine development. PMID- 18393604 TI - Rotavirus vaccine introduction in the Americas: progress and lessons learned. AB - In Latin America and the Caribbean, rotavirus causes approximately 15,000 deaths, 75,000 hospitalizations, 2 million clinic visits and 10 million cases of rotavirus diarrhea annually. Two safe vaccines are available that are effective in preventing severe illness. To date, seven countries in Latin America (Brazil, Ecuador, El Salvador, Panama, Mexico, Nicaragua and Venezuela) have introduced the vaccine. For successful rotavirus vaccine introduction, the lessons learned re-emphasize the critical need for countries to have precise plans that will ensure technical, programmatic and financial sustainability of vaccine introduction. Of these lessons learned, programmatic feasibility and financial sustainability were particularly challenging for countries that were the first to introduce a rotavirus vaccine. PMID- 18393605 TI - Feasibility of global measles eradication after interruption of transmission in the Americas. AB - Measles is one of the most infectious diseases. Before the introduction of the measles vaccine, nearly all children contracted measles. By the end of the 1980s, most countries of the world had incorporated the measles vaccine into their routine vaccination programs. Globally, some 345,000 deaths due to measles still occur every year. Eradication of measles would play an important role in improving child survival. The goal to eradicate measles from the Americas was set by the Pan-American Sanitary Conference in 1994. Progress to date has been remarkable. Measles is no longer an endemic disease in the Americas and interruption of transmission has been documented in most countries. As of December 2007, 5 years have elapsed since the detection of the last endemic case in Venezuela in November 2002. This experience demonstrates that interruption of measles transmission can be achieved and sustained over a long period of time. Global eradication should be feasible if the appropriate strategies are implemented. Even in a new paradigm in which eradication is not followed by the discontinuation of vaccination, eradication of measles should be a good investment to avoid expensive epidemics and save those children that would potentially die due to infection with the measles virus. It is not only a dream to think that we will see a world free of measles by the year 2015. PMID- 18393606 TI - Baculovirus as an expression and/or delivery vehicle for vaccine antigens. AB - The baculovirus/insect cell-expression system has proven to be a valuable tool for rapid expression of abundant recombinant proteins for research purposes and has been increasingly exploited for the production of vaccine candidates for commercial use. Furthermore, baculovirus has been discovered to be capable of efficiently transducing a wide variety of mammalian cells, thus leading to the emergence of baculovirus as a novel vector for in vivo and in vitro gene delivery. By incorporating a mammalian expression cassette into the viral genome and/or genetically modifying the baculovirus envelope for immunogen display, baculovirus has also been exploited recently as a vaccine expression/delivery vehicle. This review will focus primarily on past progress and recent advances with regards to employing baculovirus as an in vitro or in vivo expression/delivery vehicle for vaccine immunogens. PMID- 18393607 TI - Type I interferons as vaccine adjuvants against infectious diseases and cancer. AB - Presently, new attention is given to type I interferons (IFNs) as essential factors linking innate and adaptive immunity. Several studies provided evidence about the importance of IFN-alpha in the differentiation of the Th1 subset, in the generation and activity of cytotoxic T lymphocytes, in the enhancement of a primary antibody response and in the activation of dendritic cells. Owing to their immunomodulatory properties, type I IFNs can represent good candidates to be used as adjuvants for vaccination. In the present review, we summarize recent studies in humans and in animal models, suggesting a possible application of type I IFNs as adjuvants for the development of more effective vaccines against infectious diseases and cancer. PMID- 18393608 TI - Heat-shock protein-based vaccines for cancer and infectious disease. AB - Almost 60 years ago, the pioneering work of George Klein and others showed that cancers could be made targets for the immune system. Identification of the tumor targets, known as tumor antigens, became a focus in cancer biology that led to the discovery of the immunological properties of heat-shock proteins (HSPs) in 1986 by Pramod Srivastava and colleagues. Since then, the use of HSPs in the therapeutics of cancer and infectious disease in several clinical trials has been guided by our understanding of the role and effects of HSPs in adaptive and innate immune responses, investigated primarily in mice. This review will highlight the immunological properties of HSPs as we understand them today and review the clinical work on human cancers. Several Phase I and II clinical trials in different types of cancer that have been completed, as well as ongoing Phase III trials, will be summarized. PMID- 18393610 TI - Plant receptor-like serine threonine kinases: roles in signaling and plant defense. AB - Plants are hosts to a wide array of pathogens from all kingdoms of life. In the absence of an active immune system or combinatorial diversifications that lead to recombination-driven somatic gene flexibility, plants have evolved different strategies to combat both individual pathogen strains and changing pathogen populations. The receptor-like kinase (RLK) gene-family expansion in plants was hypothesized to have allowed accelerated evolution among domains implicated in signal reception, typically a leucine-rich repeat (LRR). Under that model, the gene-family expansion represents a plant-specific adaptation that leads to the production of numerous and variable cell surface and cytoplasmic receptors. More recently, it has emerged that the LRR domains of RLK interact with a diverse group of proteins leading to combinatorial variations in signal response specificity. Therefore, the RLK appear to play a central role in signaling during pathogen recognition, the subsequent activation of plant defense mechanisms, and developmental control. The future challenges will include determinations of RLK modes of action, the basis of recognition and specificity, which cellular responses each receptor mediates, and how both receptor and kinase domain interactions fit into the defense signaling cascades. These challenges will be complicated by the limited information that may be derived from the primary sequence of the LRR domain. The review focuses upon implications derived from recent studies of the secondary and tertiary structures of several plant RLK that change understanding of plant receptor function and signaling. In addition, the biological functions of plant and animal RLK-containing receptors were reviewed and commonalities among their signaling mechanisms identified. Further elucidated were the genomic and structural organizations of RLK gene families, with special emphasis on RLK implicated in resistance to disease and development. PMID- 18393611 TI - Post-transcriptional gene silencing in the root system of the actinorhizal tree Allocasuarina verticillata. AB - In recent years, RNA interference has been exploited as a tool for investigating gene function in plants. We tested the potential of double-stranded RNA interference technology for silencing a transgene in the actinorhizal tree Allocasuarina verticillata. The approach was undertaken using stably transformed shoots expressing the beta-glucuronidase (GUS) gene under the control of the constitutive promoter 35S; the shoots were further transformed with the Agrobacterium rhizogenes A4RS containing hairpin RNA (hpRNA) directed toward the GUS gene, and driven by the 35S promoter. The silencing and control vectors contained the reporter gene of the green fluorescent protein (GFP), thus allowing a screening of GUS-silenced composite plantlets for autofluorescence. With this rapid procedure, histochemical data established that the reporter gene was strongly silenced in both fluorescent roots and actinorhizal nodules. Fluorometric data further established that the level of GUS silencing was usually greater than 90% in the hairy roots containing the hairpin GUS sequences. We found that the silencing process of the reporter gene did not spread to the aerial part of the composite A. verticillata plants. Real-time quantitative polymerase chain reaction showed that GUS mRNAs were substantially reduced in roots and, thereby, confirmed the knock-down of the GUS transgene in the GFP(+) hairy roots. The approach described here will provide a versatile tool for the rapid assessment of symbiotically related host genes in actinorhizal plants of the Casuarinaceae family. PMID- 18393612 TI - A putative MAP kinase kinase kinase, MCK1, is required for cell wall integrity and pathogenicity of the rice blast fungus, Magnaporthe oryzae. AB - Insertional mutagenesis of Magnaporthe oryzae led to the identification of MCK1, a pathogenicity gene predicted to encode mitogen-activated protein kinase kinase kinase (MAPKKK) homologous to BCK1 in Saccharomyces cerevisiae. Targeted disruption of MCK1 resulted in the fungus undergoing autolysis and showing hypersensitivity to cell-wall-degrading enzyme. The mck1 produced significantly reduced numbers of conidia and developed appressoria in a slightly retarded manner compared with the wild type. Appressorium of the mck1 mutant was unable to penetrate into plant tissues, thereby rendering the mutant nonpathogenic. Cytorrhysis assay and monitoring of lipid mobilization suggested that the appressorial wall was altered, presumably affecting the level of turgor pressure within appressorium. Furthermore, the mck1 mutant failed to grow inside plant tissue. Complementation of the mutated gene restored its ability to cause disease symptoms, demonstrating that MCK1 is required for fungal pathogenicity. Taken together, our results suggest that MCK1 is an MAPKKK involved in maintaining cell wall integrity of M. oryzae, and that remodeling of the cell wall in response to host environments is essential for fungal pathogenesis. PMID- 18393613 TI - api, A novel Medicago truncatula symbiotic mutant impaired in nodule primordium invasion. AB - Genetic approaches have proved to be extremely useful in dissecting the complex nitrogen-fixing Rhizobium-legume endosymbiotic association. Here we describe a novel Medicago truncatula mutant called api, whose primary phenotype is the blockage of rhizobial infection just prior to nodule primordium invasion, leading to the formation of large infection pockets within the cortex of noninvaded root outgrowths. The mutant api originally was identified as a double symbiotic mutant associated with a new allele (nip-3) of the NIP/LATD gene, following the screening of an ethylmethane sulphonate-mutagenized population. Detailed characterization of the segregating single api mutant showed that rhizobial infection is also defective at the earlier stage of infection thread (IT) initiation in root hairs, as well as later during IT growth in the small percentage of nodules which overcome the primordium invasion block. Neither modulating ethylene biosynthesis (with L-alpha-(2-aminoethoxyvinylglycine or 1 aminocyclopropane-1-carboxylic acid) nor reducing ethylene sensitivity in a skl genetic background alters the basic api phenotype, suggesting that API function is not closely linked to ethylene metabolism or signaling. Genetic mapping places the API gene on the upper arm of the M. truncatula linkage group 4, and epistasis analyses show that API functions downstream of BIT1/ERN1 and LIN and upstream of NIP/LATD and the DNF genes. PMID- 18393614 TI - Citrate uptake into Pectobacterium atrosepticum is critical for bacterial virulence. AB - To analyze whether metabolite import into Pectobacterium atrosepticum cells affects bacterial virulence, we investigated the function of a carrier which exhibits significant structural homology to characterized carboxylic-acid transport proteins. The corresponding gene, ECA3984, previously annotated as coding for a Na(+)/sulphate carrier, in fact encodes a highly specific citrate transporter (Cit1) which is energized by the proton-motive force. Expression of the cit1 gene is stimulated by the presence of citrate in the growth medium and is substantial during growth of P. atrosepticum on potato tuber tissue. Infection of tuber tissue with P. atrosepticum leads to reduced citrate levels. P. atrosepticum insertion mutants, lacking the functional Cit1 protein, did not grow in medium containing citrate as the sole carbon source, showed a substantially reduced ability to macerate potato tuber tissue, and did not provoke reduced citrate levels in the plant tissue upon infection. We propose that citrate uptake into P. atrosepticum is critical for full bacterial virulence. PMID- 18393615 TI - Two Xanthomonas extracellular polygalacturonases, PghAxc and PghBxc, are regulated by type III secretion regulators HrpX and HrpG and are required for virulence. AB - Xanthomonas campestris pv. campestris, the causal agent of black rot disease, produces a suite of extracellular cell-wall degrading enzymes (CWDE) that are involved in bacterial virulence. Polygalacturonase (PG) is an important CWDE and functions to degrade the pectic layers of plant cell walls. Although previous studies have documented the virulence functions of PG in Erwinia and Ralstonia species, the regulation of PG genes still needs to be elucidated. In this study, we identified two novel PG genes (pghAxc and pghBxc) encoding functional PG from X. campestris pv. campestris 8004. The expressions of these two PG genes are regulated by the type III secretion regulators HrpX and HrpG and the global regulator Clp. These PG genes could be efficiently induced in planta and were required for the full virulence of X. campestris pv. campestris to Arabidopsis. In addition, these PG were confirmed to be secreted via the type II secretion system in an Xps-dependent manner. PMID- 18393616 TI - An oleic acid-mediated pathway induces constitutive defense signaling and enhanced resistance to multiple pathogens in soybean. AB - Stearoyl-acyl carrier protein-desaturase (SACPD)-catalyzed synthesis of oleic acid (18:1) is an essential step in fatty acid biosynthesis. Arabidopsis mutants (ssi2) with reduced SACPD activity accumulate salicylic acid (SA) and exhibit enhanced resistance to multiple pathogens. We show that reduced levels of 18:1 induce similar defense-related phenotypes in soybean. A Bean pod mottle virus (BPMV)-based vector was employed to effectively silence soybean SACPDs. The silenced plants contained reduced 18:1 and increased stearic acid, developed spontaneous cell death lesions, increased SA accumulation, and constitutively expressed pathogenesis-related genes. These plants also expressed elevated levels of resistance-like genes and showed resistance to bacterial and oomycete pathogens. Exogenous application of glycerol induced similar phenotypes, mimicking the effect of silencing SACPDs in healthy soybean plants. Overexpression of a soybean SACPD increased 18:1 levels in ssi2 but not in wild type Arabidopsis plants, suggesting that the soybean enzyme was under feedback regulation similar to that of the Arabidopsis isozymes. These results suggest that soybean and Arabidopsis respond similarly to 18:1-derived cues by inducing a novel broad-spectrum resistance-conferring pathway, even though they differ significantly in their lipid biosynthetic pathways. We also demonstrate the efficacy of BPMV-induced gene silencing as a tool for functional studies in soybean. PMID- 18393617 TI - Silencing a candidate nematode effector gene corresponding to the tomato resistance gene Mi-1 leads to acquisition of virulence. AB - The Mi-1 gene in tomato confers effective resistance against several species of root-knot nematode, including Meloidogyne javanica. A strain of M. javanica that can reproduce on tomato with Mi-1 was obtained from a culture of an avirulent strain after greenhouse selection. DNA blots and amplified fragment length polymorphism (AFLP) analysis indicated that the two nematode strains are closely related. Expression patterns visualized as cDNA AFLPs were nearly identical except for a cDNA fragment, Cg-1, that was present in the avirulent strain but not in the virulent strain. DNA blots showed that Cg-1 corresponds to a member of a small gene family with one or more copies missing in the virulent strain compared with the avirulent strain. Except for the presence of a histone stem loop near the 3' end of the transcript, Cg-1 shows no similarity to other sequences in GenBank. The longest open reading frame is 32 amino acids and initiates at the fourth AUG in the predicted transcript. When nematode juveniles of the Mi-1-avirulent strain were soaked in dsRNA corresponding to part of the predicted Cg-1 transcript, they produced progeny that were virulent on tomato carrying the Mi-1 gene, strongly suggesting that Cg-1 is required in the nematode for Mi-1-mediated resistance. PMID- 18393618 TI - NTH201, a novel class II KNOTTED1-like protein, facilitates the cell-to-cell movement of Tobacco mosaic virus in tobacco. AB - NTH201, a novel class II KNOTTED1-like protein gene, was cloned from tobacco (Nicotiana tabacum cv. Xanthi) and its role in Tobacco mosaic virus (TMV) infection was analyzed. Virus-induced gene silencing of NTH201 caused a delay in viral RNA accumulation as well as virus spread in infected tobacco plants. Overexpression of the gene in a transgenic tobacco plant (N. tabacum cv. Xanthi nc) infected by TMV showed larger local lesions than those of the nontransgenic plant. NTH201 exhibited no intercellular trafficking ability but did exhibit colocalization with movement protein (MP) at the plasmodesmata. When NTH201 overexpressing tobacco BY-2 cultured cells were infected with TMV, the accumulation of MP but not of viral genomic and subgenomic RNA clearly was accelerated compared with those in nontransgenic cells at an early infection period. The formation of virus replication complexes (VRC) also was accelerated in these transgenic cells. Conversely, NTH201-silenced cells showed less MP accumulations and fewer VRC formations than did nontransgenic cells. These results suggested that NTH201 might indirectly facilitate MP accumulation and VRC formation in TMV-infected cells, leading to rapid viral cell-to-cell movement in plants at an early infection stage. PMID- 18393619 TI - Host-dependent expression of Rhizobium leguminosarum bv. viciae hydrogenase is controlled at transcriptional and post-transcriptional levels in legume nodules. AB - The legume host affects the expression of Rhizobium leguminosarum hydrogenase activity in root nodules. High levels of symbiotic hydrogenase activity were detected in R. leguminosarum bacteroids from different hosts, with the exception of lentil (Lens culinaris). Transcription analysis showed that the NifA-regulated R. leguminosarum hydrogenase structural gene promoter (P(1)) is poorly induced in lentil root nodules. Replacement of the P(1) promoter by the FnrN-dependent promoter of the fixN gene restored transcription of hup genes in lentil bacteroids, but not hydrogenase activity. In the P(fixN)-hupSL strain, additional copies of the hup gene cluster and nickel supplementation to lentil plants increased bacteroid hydrogenase activity. However, the level of activity in lentil still was significantly lower than in pea bacteroids, indicating that an additional factor is impairing hydrogenase expression inside lentil nodules. Immunological analysis revealed that lentil bacteroids contain reduced levels of both hydrogenase structural subunit HupL and nickel-binding protein HypB. Altogether, results indicate that hydrogenase expression is affected by the legume host at the level of both transcription of hydrogenase structural genes and biosynthesis or stability of nickel-related proteins HypB and HupL, and suggest the existence of a plant-dependent mechanism that affects hydrogenase activity during the symbiosis by limiting nickel availability to the bacteroid. PMID- 18393620 TI - Oxalic acid is an elicitor of plant programmed cell death during Sclerotinia sclerotiorum disease development. AB - Accumulating evidence supports the idea that necrotrophic plant pathogens interact with their hosts by controlling cell death. Sclerotinia sclerotiorum is a necrotrophic ascomycete fungus with a broad host range (>400 species). Previously, we established that oxalic acid (OA) is an important pathogenicity determinant of this fungus. In this report, we describe a mechanism by which oxalate contributes to the pathogenic success of this fungus; namely, that OA induces a programmed cell death (PCD) response in plant tissue that is required for disease development. This response exhibits features associated with mammalian apoptosis, including DNA laddering and TUNEL reactive cells. Fungal mutants deficient in OA production are nonpathogenic, and apoptotic-like characteristics are not observed following plant inoculation. The induction of PCD by OA is independent of the pH-reducing abilities of this organic acid, which is required for sclerotial development. Moreover, oxalate also induces increased reactive oxygen species (ROS) levels in the plant, which correlate to PCD. When ROS induction is inhibited, apoptotic-like cell death induced by OA does not occur. Taken together, we show that Sclerotinia spp.-secreted OA is an elicitor of PCD in plants and is responsible for induction of apoptotic-like features in the plant during disease development. This PCD is essential for fungal pathogenicity and involves ROS. Thus, OA appears to function by triggering in the plant pathways responsible for PCD. Further, OA secretion by Sclerotinia spp. is not directly toxic but, more subtly, may function as a signaling molecule. PMID- 18393621 TI - Multiple activities associated with the capsid protein of satellite panicum mosaic virus are controlled separately by the N- and C-terminal regions. AB - The 17-kDa capsid protein (CP) of satellite panicum mosaic virus (SPMV) contains a distinct N-terminal arginine-rich motif (N-ARM) which is required for SPMV virion assembly and the activity of SPMV CP to promote systemic accumulation of its cognate RNA. The present study indicates that SPMV CP also is involved in SPMV RNA accumulation in inoculated leaves and that this activity is also dependent on a functional N-ARM. In addition, deletions of a C-terminal region abolish virion assembly and impair SPMV RNA accumulation in both inoculated and systemic leaves. Unlike the N-ARM mutations, substantial deletions of the SPMV CP C-terminus do not affect SPMV RNA binding activity. Interestingly, SPMV CP also binds Panicum mosaic virus genomic RNA via N-ARM-mediated CP:RNA interactions. Mutations of the N-ARM and the C-terminal regions significantly reduce SPMV CP titers and result in symptom attenuation. In contrast, virions were not associated per se with symptom exacerbation or successful SPMV RNA accumulation. The results show the existence of a correlation between N- and C-termini-mediated contributions for CP accumulation, symptom induction, defective-interfering RNA accumulation, and temperature sensitivity of SPMV RNA maintenance. The data provide further evidence that SPMV CP has multiple roles during infection, which might involve the formation of nonvirion CP:RNA complexes whose stability is controlled in a biologically relevant manner by the N- and C-termini of the CP. PMID- 18393622 TI - Evidence for transcriptional and post-translational regulation of sucrose synthase in pea nodules by the cellular redox state. AB - Nitrogen fixation (NF) in legume nodules is very sensitive to environmental constraints. Nodule sucrose synthase (SS; EC 2.4.1.13) has been suggested to play a crucial role in those circumstances because its downregulation leads to an impaired glycolytic carbon flux and, therefore, a depletion of carbon substrates for bacteroids. In the present study, the likelihood of SS being regulated by oxidative signaling has been addressed by the in vivo supply of paraquat (PQ) to nodulated pea plants and the in vitro effects of oxidizing and reducing agents on nodule SS. PQ produced cellular redox imbalance leading to an inhibition of NF. This was preceded by the downregulation of SS gene expression, protein content, and activity. In vitro, oxidizing agents were able to inhibit SS activity and this inhibition was completely reversed by the addition of dithiothreitol. The overall results are consistent with a regulation model of nodule SS exerted by the cellular redox state at both the transcriptional and post-translational levels. The importance of such mechanisms for the regulation of NF in response to environmental stresses are discussed. PMID- 18393623 TI - Transcription profiling of soybean nodulation by Bradyrhizobium japonicum. AB - Legumes interact with nodulating bacteria that convert atmospheric nitrogen into ammonia for plant use. This nitrogen fixation takes place within root nodules that form after infection of root hairs by compatible rhizobia. Using cDNA microarrays, we monitored gene expression in soybean (Glycine max) inoculated with the nodulating bacterium Bradyrhizobium japonicum 4, 8, and 16 days after inoculation, timepoints that coincide with nodule development and the onset of nitrogen fixation. This experiment identified several thousand genes that were differentially expressed in response to B. japonicum inoculation. Expression of 27 genes was analyzed by quantitative reverse transcriptase-polymerase chain reaction, and their expression patterns mimicked the microarray results, confirming integrity of analyses. The microarray results suggest that B. japonicum reduces plant defense responses during nodule development. In addition, the data revealed a high level of regulatory complexity (transcriptional, post transcriptional, translational, post-translational) that is likely essential for development of the symbiosis and adjustment to an altered nutritional status. PMID- 18393624 TI - Discovery of ADP-ribosylation and other plant defense pathway elements through expression profiling of four different Arabidopsis-Pseudomonas R-avr interactions. AB - A dissection of plant defense pathways was initiated through gene expression profiling of the responses of a single Arabidopsis thaliana genotype to isogenic Pseudomonas syringae strains expressing one of four different cloned avirulence (avr) genes. Differences in the expression profiles elicited by different resistance (R)-avr interactions were observed. A role for poly(ADP-ribosyl)ation in plant defense responses was suggested initially by the upregulated expression of genes encoding NUDT7 and poly(ADP-ribose) glycohydrolase in multiple R-avr interactions. Gene knockout plant lines were tested for 20 candidate genes identified by the expression profiling, and Arabidopsis NUDT7 mutants allowed less growth of virulent P. syringae (as previously reported) but also exhibited a reduced hypersensitive-response phenotype. Inhibitors of poly(ADP-ribose) polymerase (PARP) disrupted FLS2-mediated basal defense responses such as callose deposition. EIN2 (ethylene response) and IXR1 and IXR2 (cellulose synthase) mutants impacted the FLS2-mediated responses that occur during PARP inhibition, whereas no impacts were observed for NPR1, PAD4, or NDR1 mutants. In the expression profiling work, false-positive selection and grouping of genes was reduced by requiring simultaneous satisfaction of statistical significance criteria for each of three separate analysis methods, and by clustering genes based on statistical confidence values for each gene rather than on average fold change of transcript abundance. PMID- 18393625 TI - Genome organization and evolution of the AVR-Pita avirulence gene family in the Magnaporthe grisea species complex. AB - The avirulence (AVR) gene AVR-Pita in Magnaporthe oryzae prevents the fungus from infecting rice cultivars containing the resistance gene Pi-ta. A survey of isolates of the M. grisea species complex from diverse hosts showed that AVR-Pita is a member of a gene family, which led us to rename it to AVR-Pita1. Avirulence function, distribution, and genomic context of two other members, named AVR-Pita2 and AVR-Pita3, were characterized. AVR-Pita2, but not AVR-Pita3, was functional as an AVR gene corresponding to Pi-ta. The AVR-Pita1 and AVR-Pita2 genes were present in isolates of both M. oryzae and M. grisea, whereas the AVR-Pita3 gene was present only in isolates of M. oryzae. Orthologues of members of the AVR-Pita family could not be found in any fungal species sequenced to date, suggesting that the gene family may be unique to the M. grisea species complex. The genomic context of its members was analyzed in eight strains. The AVR-Pita1 and AVR-Pita2 genes in some isolates appeared to be located near telomeres and flanked by diverse repetitive DNA elements, suggesting that frequent deletion or amplification of these genes within the M. grisea species complex might have resulted from recombination mediated by repetitive DNA elements. PMID- 18393627 TI - Correlation between plasma Flt3-ligand concentration and hematopoiesis during G CSF-induced CD34+ cell mobilization. AB - This study aimed to correlate blood Flt3-ligand (FL) concentration with CD34(+) cell number in blood and bone marrow (BM) during granulocyte colony-stimulating factor (G-CSF) mobilization. Nonhuman primates were injected with 10 microg/kg of G-CSF (Lenograstim) daily over a period of 5 days. Daily blood sampling and repeated BM sampling showed that FL concentration before mobilization was negatively correlated to the absolute number of BM CD34(+) cells, but also to the number of G-CSF-mobilized CD34(+) cells on days 3-5 of treatment. This showed that FL concentration in the blood reflected BM status before mobilization, and suggested that this parameter could be used as a predictive indicator of G-CSF induced CD34(+) cell mobilization. PMID- 18393628 TI - Functional nicotinic and muscarinic receptors on mesenchymal stem cells. AB - Mesenchymal stem cells (MSCs) are under the control of a large number of signaling systems. In this study, the presence and functionality of the acetylcholine (ACh) signaling system in MSCs was examined. We detected the expression of choline acetyltransferase (ChAT), acetylcholinesterase (AChE), and the presence of ACh in MSCs. MSCs also expressed the nicotinic acetylcholine receptor subunits alpha 3, alpha 5, alpha 7, and the muscarinic acetylcholine receptor 2 (M2-receptor). The M2-receptor and the nicotinic alpha 7 receptor subunits were expressed on distinct subpopulations of cells, indicating differential regulation of cholinergic signaling between MSCs. Stimulation of MSCs with the nicotinic receptor agonist nicotine and the muscarinic receptor agonist muscarine induced immediate and transient increases in intracellular Ca(2+) concentration. Furthermore, muscarine had an inhibiting effect on the production of the intracellular signaling molecule cyclic adenosine 3',5' monophosphate (cAMP). The AChE inhibitor chlorpyrifos, which is widely used as an agricultural insecticide, had similar effects on intracellular Ca(2+) and cAMP in MSCs. Nicotine, muscarine, and chlorpyrifos induced the phosphorylation of extracellular signal-regulated kinases 1 and 2. This study demonstrates that several components of a cholinergic signaling system are present and functional in MSCs. Environmental compounds such as nicotine and agricultural insecticides can interfere with this system and may affect cellular processes in the MSC. PMID- 18393629 TI - Propagation of undifferentiated human embryonic stem cells with nano-liposomal ceramide. AB - Human embryonic stem (hES) cells, located on the periphery of the colonies, express the neuroectodermal markers nestin and Tuj1, suggesting a prematurely differentiated subgroup of cells. Here, we report that ceramide, a bioactive sphingolipid, selectively eliminates hES cells differentially expressing nestin and Tuj1. In contrast, undifferentiated cells are resistant to the apoptotic effects of ceramide. Ceramide-resistant hES cells express higher levels of the messenger RNA for ceramide-metabolizing enzymes that convert ceramide into pro mitogenic metabolites. Based on these findings, we conducted long-term studies to determine whether liposomal ceramide can be used to maintain undifferentiated hES cells free of feeder cells. We continuously cultured hES cells on matrigel for 4 months with liposomal ceramide in a feeder cell-free system. Human ES cells treated with liposomal ceramide maintained their pluripotent state as determined by in vivo and in vitro differentiation studies and contained no chromosomal abnormalities. In conclusion, our findings suggest that exposure to ceramide provides a viable strategy to prevent premature hES cell differentiation and to maintain pluripotent stem cell populations in the absence of feeder cells. PMID- 18393630 TI - Non-invasive longitudinal tracking of human amniotic fluid stem cells in the mouse heart. AB - Human stem cells from various sources have potential therapeutic applications. The clinical implementation of these therapies introduces the need for methods of noninvasive tracking of cells. The purpose of this study was to evaluate a high resolution magnetic resonance imaging (MRI) technique for in vivo detection and tracking of superparamagnetic micron sized iron oxide particle (MPIO)-labeled human amniotic fluid stem (hAFS) cells injected in the mouse heart. Because of the small subject size, MR signal and resolution of the in vivo MRI were increased using strong gradients, a 7.0 Tesla magnet, and an ECG and respiratory gated gradient echo sequence. MRI images of mouse heart were acquired during a 4 week course of this longitudinal study. At the end of the study, histological analysis was used to correlate cell localization with the MRI results. Introduction of MPIOs into hAFS had no significant effect upon cell proliferation and differentiation. Results of flow cytometry analysis indicated that hAFS cells remained labeled for up to 4 weeks. MRI of MPIO-labeled hAFS cells injected in agarose gels resulted in significant hypointense regions. Labeled hAFS cells injected into mouse hearts produced hypointense regions in the MR images that could be detected 24 hours and 7, 14, 21 and 28 days post injection. The co localization of labeled cells within the hypointense regions was confirmed by histological analysis. These results indicate that high resolution MRI can be used successfully for noninvasive longitudinal tracking of hAFS cells injected in the mouse heart. The potential utility of this finding is that injected stem cells can be tracked in vivo and might serve to monitor cell survival, proliferation and integration into myocardial tissue. PMID- 18393631 TI - Sphingosine-1-phosphate mediates transcriptional regulation of key targets associated with survival, proliferation, and pluripotency in human embryonic stem cells. AB - Human embryonic stem cells (hESCs) replicate in vitro by the process of self renewal, whilst maintaining their pluripotency. Understanding the pathways involved in the regulation of this process will assist in developing fully defined conditions for the robust proliferation of hESCs necessary for therapeutic applications. We previously demonstrated that sphingosine-1-phosphate (S1P) plays an important role in survival and proliferation of hESCs. and here the key signaling pathways and downstream targets of S1P were investigated in a representative cell line (Shef 4). A significant rise in ERK1/2 activation with S1P treatment was witnessed in hESCs maintained on murine embryonic fibroblasts (MEFs) exhibiting significantly higher levels of active ERK1/2 than those grown on Matrigel. RT-PCR and microarray analysis of micro-dissected, non differentiated hESC revealed 1049 differentially expressed genes in S1P treated preparations compared with controls (n = 3). S1P regulated apoptosis through several BCL-2 family members, including BAX and BID, with increased expression of cell cycle progression genes associated with proliferation of hESC cultures. S1P treatment also increased expression of cell adhesion genes specifically cadherins and integrins. However, gene expression associated with pluripotency was decreased with S1P treatment indicating that an increased rate of hESC turnover (higher proliferation and lower apoptosis) may be balanced by an increased susceptibility to differentiate. PMID- 18393632 TI - Control of early fate decisions in human ES cells by distinct states of TGFbeta pathway activity. AB - The mechanisms controlling self-renewal versus lineage commitment in human embryonic stem (hES) cells are not well understood. Nonetheless, current knowledge suggests a crucial role for TGFbeta signaling in controlling these early fate decisions. We have investigated the effects of TGFbeta pathway activation and inhibition on gene expression in hES cells. Our data reveal that SMAD 2/3 signaling directly supports NANOG expression, while SMAD 1/5/8 activation moderately represses SOX2. In addition, genes encoding key developmentally relevant signaling molecules and transcription factors appear to be immediately downstream of SMAD 1/5/8 signaling, or require both SMAD 1/5/8 and 2/3 activation, or inactivation of TGFbeta signaling for their induction. Defined stimulation/inhibition of the two TGFbeta branches appeared to control early fate decisions in accordance with these downstream transcriptional effects. Our results therefore help to better understand how pluripotency is mediated at the transcriptional level. PMID- 18393634 TI - Comparative analysis of mesenchymal stem cells from bone marrow, cartilage, and adipose tissue. AB - Mesenchymal stem cells (MSCs) isolated from bone marrow (BM), cartilage, and adipose tissue (AT) possess the capacity for self-renewal and the potential for multilineage differentiation, and are therefore perceived as attractive sources of stem cells for cell therapy. However, MSCs from these different sources have different characteristics. We compared MSCs of adult Sprague Dawley rats derived from these three sources in terms of their immunophenotypic characterization, proliferation capacity, differentiation ability, expression of angiogenic cytokines, and anti-apoptotic ability. According to growth curve, cell cycle, and telomerase activity analyses, MSCs derived from adipose tissue (AT-MSCs) possess the highest proliferation potential, followed by MSCs derived from BM and cartilage (BM-MSCs and C-MSCs). In terms of multilineage differentiation, MSCs from all three sources displayed osteogenic, adipogenic, and chondrogenic differentiation potential. The result of realtime RT-PCR indicated that these cells all expressed angiogenic cytokines, with some differences in expression level. Flow cytometry and MTT analysis showed that C-MSCs possess the highest resistance toward hydrogen peroxide -induced apoptosis, while AT-MSCs exhibited high tolerance to serum deprivation-induced apoptosis. Both AT and cartilage are attractive alternatives to BM as sources for isolating MSCs, but these differences must be considered when choosing a stem cell source for clinical application. PMID- 18393636 TI - Neuron-like differentiation and selective ablation of undifferentiated embryonic stem cells containing suicide gene with Oct-4 promoter. AB - In vivo transplantation of undifferentiated embryonic stem (ES) cells can produce teratomas with uncontrolled cell proliferation. Although ES cells may be attractive candidates for human cell-replacement therapy in the future, the major limitation of its application to the therapy is teratoma formation. In the present study, ES cells containing herpes simplex virus-thymidine kinase (HSV-tk) transgene for a suicide gene expression under the control of the Oct-4 promoter was used for ablation of undifferentiated ES cells, which may produce teratomas, using three-dimensional cell culture system allowing a multilayer cell construct. Selective ablation of undifferentiated ES cells expressing HSV-tk gene under the control of Oct-4 promoter was achieved by ganciclovir treatment. Surviving ES cells after ganciclovir treatment expressed several neuron-associated markers such as synaptophysin, beta-tubulin, vesicular glutamate transporter 1, syntaxin, protein kinase C and glial fibrillary acidic protein (GFAP) but not Oct-4. Coexpression of synaptophysin as a marker of neuronal synapse and GFAP as that of glial fibers in the surviving ES cells revealed finely structured neuronal network. Furthermore, decrease of Ki-67 proliferative index was detected in the surviving ES cells. In conclusion, selective ablation of undifferentiated ES cells by a suicide gene decreases proliferative activity and induces neuron-like differentiation in ES cells. PMID- 18393637 TI - Migration and differentiation of human umbilical cord stem cells after heart injury in chicken embryos. AB - Here we have analyzed the behavior and fate of stem cells from human umbilical cord blood (scHUCBs) when grafted into the myocardial wall of normal and damaged hearts of chicken embryos. We started by characterizing the scHUCBs before grafting and we found that they express precardiogenic genes including Nkx2.5, GATA4, MEF-2, and SERCA2a together with undifferentiation markers as CD34 or c kit. In grafting experiments using scHUCBs labeled with DiI we observed that these cells were not rejected by the host and survived when implanted in chicken hearts, being able to migrate through the myocardial wall. By 3 days after grafting we found labeled cells with morphological characters of myocardiocytes in concordance with the identification of the expression of human genes for myosin light chain 2a (Mlc2a) and myosin heavy chain-beta (Mhc beta) in the chicken heart. When a small injury was applied to the heart wall, grafted scHUCBs were vigorously attracted by the damaged myocardium. This directed migration was only sustained for 12 h after injury, time period required for healing of the damaged heart wall. The rate of myocardial differentiation of scHUCBs in damaged hearts was not significantly increased with respect to that found when implanted in healthy hearts. However, we found stimulation of endothelial differentiation in injured hearts deduced by the increased expression of human genes for platelet endothelial cell-adhesion molecule 1 or vascular endothelial growth factor receptor 2 and the presence of DiI-labeled endothelial cells. Together all these findings support the embryonic chicken heart as a feasible model for experimentation in stem cell therapy and emphasize the relevance of the physiological conditions of the myocardial host tissue for engraftment and differentiation of exogenously applied scHUCBs. PMID- 18393635 TI - Epigenetic regulation of mammalian stem cells. AB - Two critical properties of stem cells are self-renewal and multipotency. The maintenance of their "stemness" state and commitment to differentiation are therefore tightly controlled by intricate molecular networks. Epigenetic mechanisms, including DNA methylation, chromatin remodeling and the noncoding RNA mediated process, have profound regulatory roles in mammalian gene expression. Recent studies have shown that epigenetic regulators are key players in stem cell biology and their dysfunction can result in human diseases such as cancer and neurodevelopmental disorders. Here, we review the recent evidences that advance our knowledge in epigenetic regulations of mammalian stem cells, with focus on embryonic stem cells and neural stem cells. PMID- 18393638 TI - Human dental pulp stem cells differentiate into neural crest-derived melanocytes and have label-retaining and sphere-forming abilities. AB - Adult tissues contain highly proliferative, clonogenic cells that meet criteria of multipotent stem cells and are potential sources for autologous reparative and reconstructive medicine. We demonstrated that human dental pulp contains self renewing human dental pulp stem cells (hDPSCs) capable of differentiating into mesenchymal-derived odontoblasts, osteoblasts, adipocytes, and chondrocytes and striated muscle, and interestingly, also into non-mesenchymal melanocytes. Furthermore, we showed that hDPSC cultures include cells with the label-retaining and sphere-forming abilities, traits attributed to multipotent stem cells, and provide evidence that these may be multipotent neural crest stem cells. PMID- 18393640 TI - From fluorescence nanoscopy to nanoscopic medicine. PMID- 18393639 TI - Proteomic characterization of the conditioned media produced by the visceral endoderm-like cell lines HepG2 and END2: toward a defined medium for the osteogenic/chondrogenic differentiation of embryonic stem cells. AB - Visceral endoderm (VE) is an extraembryonic cell layer formed before gastrulation that secretes critical factors involved in embryonic development with inductive effects on embryonic stem cell (ESC) differentiation. We utilized the conditioned media (CM) from the VE-like cell lines, HepG2 and END2, to enhance lineage specific differentiation of murine ESCs (mESCs) toward the osteogenic lineage. Previously, we have demonstrated that use of the HepG2-CM resulted in the efficient osteogenic/chondrogenic differentiation of mESCs without embryoid body (EB) formation. In this study, we demonstrate, for the first time, the osteogenic inducing activity of END2-CM, suggesting a potential shared protein profile between HepG2- and END2-CM. To identify the active factors in the CM, proteomic analysis using differential gel electrophoresis coupled with matrix-assisted laser desorption/ionization was performed on the two CM, resulting in six proteins being established to be present in both CM, including ones that may function on the epithelial-mesenchymal transition (EMT). Our results represent the first study on the VE-like cell line secretome and provide information on VE proteins identifying possible candidate proteins to be used for mesoderm-specific osteogenic differentiation. PMID- 18393642 TI - Upconverting nanoparticles as nanotransducers for photodynamic therapy in cancer cells. AB - BACKGROUND: Photodynamic therapy (PDT) involves killing of diseased cells by excitation of photosensitizer chemicals with high-energy light to produce cytotoxic oxygen species from surrounding dissolved oxygen. However, poor tissue penetration of high-energy light and hydrophobic photosensitizers limits the effectiveness to superficial pathologies. Upconversion phosphor nanoparticles convert low-energy radiation to higher-energy emissions. AIM: To create upconverting 'nanotransducers' to enable PDT in deep tissues. RESULTS: Monodisperse, 50 nm PEI/NaYF(4):Yb(3+),Er(3+) nanoparticles producing green/red emission on near-infrared (NIR) excitation were targeted to folate receptors on human colon cancer cells and imaged with high signal-to-background ratio. It was demonstrated that these particles could be excited after deep intramuscular injection in rats. On NIR excitation, the particles, modified with zinc phthalocyanin photosensitizer, released singlet oxygen and, after targeted binding to cancer cells, resulted in significant cell destruction. CONCLUSION: Potential clinical use of these nanoparticles includes imaging and PDT of cancer in deep tissues. PMID- 18393643 TI - Local anesthesia with 20-mL prilocaine infiltration: the ultimate point for analgesia during shockwave lithotripsy? AB - PURPOSE: The effectiveness of 10 mL and 20 mL local prilocaine infiltration for analgesic purposes during shockwave lithotripsy (SWL) was compared, and the differences in pain intensity and need for additional analgesics were defined. PATIENTS AND METHODS: One hundred and twenty-one patients between the ages of 16 and 69 with kidney stones were randomly separated into two groups. The 60 patients in group 1 received 10 mL prilocaine subcutaneously and deep lumbar infiltration into the area 1 cm below the twelfth rib 2 minutes before SWL. The 61 patients in group 2 received 20 mL subcutaneously and deep lumbar infiltration of prilocaine to the same locale, again 2 minutes before the process. All the patients received one session of SWL. For measurement of pain intensity, the 0 to 100 mm visual analog scale (VAS) was used at 1 minute and every 10 minutes during the process. Intravenous fentanyl was administered to patients who had steady or increasing intensity of pain during the process. RESULTS: VAS values were statistically significantly higher in patients in group 1 compared with patients in group 2 during the process. While 40 patients received additional analgesia in group 1, only three patients in group 2 needed additional analgesia. CONCLUSION: Use of 20 mL subcutaneous and deep lumbar prilocaine infiltration during SWL for analgesic purposes is recommended as an efficient, cost-effective, anesthetist independent, and reliable method. PMID- 18393644 TI - Impact of percutaneous nephrolithotomy on estimated glomerular filtration rate in patients with chronic kidney disease. AB - BACKGROUND AND PURPOSE: We investigated the impact of percutaneous renal procedures on estimated glomerular filtration rate (GFR) of patients with chronic kidney disease (CKD). PATIENTS AND METHODS: The GFRs of adult patients were calculated using the Modification of Diet in Renal Disease formula, and the patients were staged according to the Kidney Disease Outcome Quality Initiative CKD classification system. The study included 185 patients with preoperative GFR values less than 60 mL/min/1.73 m(2). The impact of percutaneous nephrolithotomy (PCNL) on GFR was analyzed by comparing the preoperative GFR with the GFR before discharge and at postoperative month 3. RESULTS: Patients with CKD had a significant increase in the GFR after the procedure. In postoperative month 3, the mean GFR was more than 60 mL/min/1.73 m(2) in 25% of the patients with CKD and less than 60 mL/min/1.73 m(2) in 75%. While all patients with stage 5 CKD improved to better stages, some other patients' conditions declined to stage 5 from better stages at the end of postoperative month 3. No patient needed dialysis. The presence of urinary tract infections tended to affect GFR negatively. CONCLUSION: Estimated GFR, as a better indicator of renal function, is significantly affected by the PCNL procedure. While significant improvement was observed in late-stage patients with CKD, unexpected deterioration could occur in patients at earlier stages. PMID- 18393645 TI - High-frequency oscillatory ventilatory support during CT-guided percutaneous cryotherapy of renal masses. AB - PURPOSE: Computed tomography (CT)-guided percutaneous procedures are often made more difficult due to the movement of the kidney during respiration. Here we examine the use of high-frequency oscillatory ventilation (HFOV), which eliminates the movement of the kidney, potentially making cryoprobe access to the kidney simpler and possibly more efficient. METHODS: We compared seven CT-guided percutaneous procedures using a single cryoprobe and either standard mechanical ventilation (MV) (n=4) or HFOV (n=3). The variables studied included: total time of patient intubation, operative time, overall duration of interventional radiology (IR) suite time, change in hematocrit, narcotic use, and complications. The ease of the procedure was rated on a subjective scale from 1 to 3. RESULTS: The total intubation time remained nearly identical at 210 minutes for HFOV and 208 minutes for MV, but surgeon procedural time decreased by 31 minutes in the HFOV group (HFOV=99 minutes and MV=130 minutes) (P=0.40). Total IR time was 225 minutes for HFOV compared to 212 minutes for the MV group (P=0.63). There were no significant differences in the postoperative hematocrit, creatinine, or narcotic use between the two groups. There were no complications related to the procedure or anesthesia in either group. Both urology attending physicians and the interventional radiologist noted that the procedure seemed easier with HFOV. CONCLUSION: HFOV may shorten the actual procedural time required to perform cryoablation, likely due to the elimination of renal movement during the procedure, thereby facilitating targeting and access to the renal mass. In this initial experience, patients tolerated HFOV without incident, and the operating surgeons found it easier to perform the procedures. PMID- 18393646 TI - Unilateral idiopathic adrenal hematomas with a preoperative diagnosis of indeterminate adrenal tumors. AB - PURPOSE: To identify the clinical and radiologic features of unilateral adrenal hematomas in adults. PATIENTS AND METHODS: From our database of 75 patients who underwent adrenalectomies at our institution, we identified 3 patients who underwent adrenalectomy for adrenal tumors. The pathology report confirmed adrenal hematomas with no evidence of neoplasm. RESULTS: The patients consisted of a man presenting with abdominal pain, another man presenting with persistent hypertension, and a woman presenting with hypertension, headache, palpitations, and diaphoresis. All patients were anemic, receiving aspirin, and had previous abdominal surgeries under general anesthesia. CONCLUSION: Unilateral adrenal hematomas sometimes are indistinguishable radiologically from neoplasms. Surgery is then necessary to distinguish a hematoma from a hemorrhagic adrenal tumor. PMID- 18393647 TI - Vessel length following laparoscopic donor nephrectomy: impact of vascular ligation technique on allograft vessel length. AB - PURPOSE: A variety of techniques have been used to secure the renal artery and vein during laparoscopic donor nephrectomy. The purpose of this study is to compare the amount of vessel length lost when the artery and vein are secured with four different techniques. METHODS: A model was constructed to simulate a left laparoscopic donor nephrectomy. In this model vessel length lost was determined when veins were secured using polymer locking (PL) clips, the endo-GIA stapling device, and the endo-TA stapling device. Arterial length lost was determined for the same three techniques, as well as securing the artery with titanium (Ti) clips. RESULTS: The mean arterial length lost for the PL clips, Ti clips, endo-TA, and endo-GIA stapling devices was 6.2, 6.3, 9.8, and 10.0 mm, respectively. Both clip types produced less loss of arterial length than both types of stapling devices (P<0.001), and there was no difference between the two types of stapling devices (P=0.73) or clips (P=0.85). The mean venous length lost for the PL clip, endo-GIA, and endo-TA stapling devices was 5.7, 10.1, and 9.4 mm, respectively. The PL clips resulted in significantly less vessel loss compared to both stapling devices (P<0.001), and there was no difference between the two stapling devices (P=0.40). CONCLUSIONS: Both types of clips resulted in longer graft arterial lengths compared to both stapling devices. PL clips resulted in longer graft vein length compared to the two stapling devices. The endo-TA stapling device was limited in this model by its inability to articulate. PMID- 18393648 TI - Risk stratification for positive lymph nodes in prostate cancer. AB - PURPOSE: To evaluate the risk of positive lymph nodes using preoperative clinical parameters. METHODS AND MATERIALS: We reviewed our prospectively collected database for all patients who received RRP and PLND between January 1993 and November 2005 as primary therapy for prostate cancer. We excluded patients who had hormonal ablation or radiation therapy prior to surgery and patients with missing PSA, clinical stage, or biopsy Gleason score. We evaluated risk for nodal disease using the following definitions: low risk: PSA or=T(2b), or Gleason score of 7; and high risk: PSA >or=20 ng/mL, or clinical stage >or=T(2c), or Gleason score >or=8. Logistic regression was used to determine the association between the risk groups and pathologic lymph node involvement, and a receiver operating characteristics (ROC) curve was constructed to evaluate the performance of the stratification scheme in detecting nodal disease. RESULTS: A total of 760 patients with 43 (5.7%) patients with node positive disease were available for analysis. Risk classification was significantly associated with positive nodes (P<0.001), even after controlling for year of surgery and age. The area under the ROC curve was 0.77 (95% CI: 0.69, 0.83). Omitting PLND in the low-risk group would have spared 368 (49.2%) of the entire cohort with a false-negative rate of 5/369 (1.3%) for the low-risk group, and 5/760 (0.7%) for the entire cohort. Sensitivity was 88.4%, and negative predictive value was 98.7%. CONCLUSION: Patients can be risk stratified for node positive disease and potentially excluded from lymphadenectomy with minimal risk. PMID- 18393649 TI - Sample size in disease management program evaluation: the challenge of demonstrating a statistically significant reduction in admissions. AB - Prior to implementing a disease management (DM) strategy, a needs assessment should be conducted to determine whether sufficient opportunity exists for an intervention to be successful in the given population. A central component of this assessment is a sample size analysis to determine whether the population is of sufficient size to allow the expected program effect to achieve statistical significance. This paper discusses the parameters that comprise the generic sample size formula for independent samples and their interrelationships, followed by modifications for the DM setting. In addition, a table is provided with sample size estimates for various effect sizes. Examples are described in detail along with strategies for overcoming common barriers. Ultimately, conducting these calculations up front will help set appropriate expectations about the ability to demonstrate the success of the intervention. PMID- 18393650 TI - Economic evaluation of an intensified disease management system for patients with type 2 diabetes. AB - We evaluated the effect of a disease management (DM) program on adherence with recommended laboratory tests, health outcomes, and health care expenditures for patients with type 2 diabetes. The study was a natural experiment in a primary care setting in which the intervention was available to 1 group and then compared to the experience of a matched control group. Univariate analysis and difference in differences analysis were used to test for any significant differences between the 2 groups following a 12-month intervention period. A payer perspective was used to estimate the health care cost consequences based on hospital and physician utilization weighted by Medicare prices. The results were nonsignificant at the .10 level, except for compliance with recommended tests, which showed significant results in the univariate analysis. The intervention increased compliance with testing for HbA1c, microalbuminuria, and lipids, and decreased HbA1c value and the percent of patients with HbA1c >or=9.5%. The point estimates showed small reductions in health care cost; only reductions in costs for office visits were significant at the .10 level. We concluded that while there were signs of improvement in adherence to testing, the low effectiveness may be attributed to existing diabetes management activities in this primary care setting, high compliance rates for testing at the beginning of the study, and a steep learning curve for this complex, information-technology-based DM system. The study raises questions about the incremental gains from complex systems approaches to DM and illustrates a rigorous method to assess DM programs under "real-world" conditions, with control for possible selection bias. PMID- 18393651 TI - Health information technology for the disease management provider. PMID- 18393652 TI - The role of incentives in the improvement of health. PMID- 18393653 TI - Oxidative stress in patients with acute heart failure. AB - Oxidative stress (OS) is a keystone in the pathology of the ischemia reperfusion sequence (acute coronary syndromes, cardiac surgery, transplantation). In heart failure, the implication of OS is less understood. This study was intended to evaluate OS in acute heart failure. Criteria for inclusion were consecutive patients hospitalized in our cardiology department for a first pulmonary edema that revealed a dilated cardiomyopathy (DCM). Exclusion criteria included known cardiomyopathy, smoker, acute coronary syndrome, and treatment with angiotensin converting enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARAII). OS was evaluated in blood samples: thiobarbituric acid-reactive substances (TBARS), total antioxidant status (TAS), plasma alpha-tocopherol, vitamin A, and beta-carotene. Standard biochemical parameters including CRP, fibrinogen, lipid, and creatinine were assayed. Ten patients (80% men, mean age 55.3 +/- 7.9 years) were included and followed during a 6 month period. The etiologies of DCM were alcohol (n = 3), anti-cancer drugs (n = 2), valvulopathies (n = 2), or idiopathic (n = 3). In acute heart failure, TBARS were elevated (1.69 micromol/L; normal value 0.6-4.2 micromol/L) and TAS status was decreased (0.96 mmol/L; normal value 1.3-1.9 pmol/L). OS was more important when patients had atrial or ventricular arrhythmia. Nevertheless, liposoluble antioxidant parameters (beta-carotene, vitamin A, alpha-tocopherol) had a usual value. At the term of the follow-up, patients returned to a stable condition, OS markers revealed normal values, and every Holter ECG showed no supraventricular or ventricular arrhythmias. In acute heart failure, oxygen-free radicals are increased. We thus hypothetized that a modification in OS could be responsible for arrhythmias and complications of acute heart failure. PMID- 18393654 TI - Making SENSE: strategies for engineering negligible senescence evolutionarily. AB - Thirty years ago, in 1977, few biologists thought that it would be possible to increase the maximum life span characteristic of each species over the variety of environmental conditions in which they live, whether in nature or in the laboratory. But the evolutionary theory of aging suggested otherwise. Accordingly, experiments were performed with fruit flies, Drosophila melanogaster, which showed that manipulation of the forces of natural selection over a number of generations could substantially slow the rate of aging, both demographically and physiologically. After this first transgression of the supposedly absolute limits to life extension, it was suggested that mammals too could be experimentally evolved to have greater life spans and slower aging. And further, it was argued that such postponed-aging mammals could be used to reverse engineer a slowing of human aging. The subsequent discovery and theoretical explanation of mortality-rate plateaus revealed that aging was not due to the progressive physiological accumulation of damage. Instead, aging is now understood by evolutionary biologists to arise from a transient fall in age specific adaptation, a fall that does not necessarily proceed toward ineluctable death. This implies that SENS must be based on re-tuning adaptation, not repairing damage. As evolutionary manipulation of model organisms shows us how adaptation can be focused on engineering negligible senescence, there are thus both scientific and practical reasons for making SENS evolutionary; that is making SENSE. PMID- 18393655 TI - Role of environmental and genetic factor interaction in age-related disease development: the gastric cancer paradigm. AB - The association of Helicobacter pylori (Hp) infection with gastric cancer is well known and might be considered a paradigmatic example of the role that interaction among environmental factors and individual background might play in inducing age associated disease. To evaluate the role of interaction of Hp infection with genetic background, gastric cancer and chronic gastritis patients as well as random selected controls were typed for five inflammation-related polymorphisms of IL-1 and IL-10 cytokine genes. No association among IL-10 or IL-1 variants with an increased risk of gastric cancer was found, whereas an Hp-independent association of IL-1beta -511T positive genotypes to an increased risk of chronic gastritis was found (Hp-/511T+ OR 1.89, 95% CI: 1.01-3.54; Hp+/-511T+ OR 1.83, 95% CI: 1.05-3.19). Stratification of gastric cancer group according to Hp infection does not allow finding a statistically significant association of Hp+ to the higher histological grading (G3) of gastric cancer (OR 1.54, 95% CI: 0.46 5.11). Our findings seem to confirm that cytokine genetic variants might contribute to determining the background for inflammaging in which H. pylori infection might facilitate cancer development. PMID- 18393656 TI - Characterizing endothelial cells derived from the murine embryonic stem cell line CCE. AB - Embryonic stem cells (ESC) are defined by two main properties of self-renewal and their multipotency to differentiate into virtually all cell types of the body, including endothelial cells. ESCs have been widely regarded as an unlimited source of cells in regeneration medicine and also an ideal in vitro model to investigate complex developmental processes. Here, we report a simple and efficient in vitro model to derive a nearly pure population of endothelial cells from a murine ESC line. CCE ES cells are exposed to alpha-MEM medium containing 10% FBS for 4 days and then cultured in endothelial basal-2 medium containing vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF), epidermal growth factor (EGF), and 2% FBS for 42 days. The cells acquired a relatively uniform endothelial cell morphology and were able to propagate and expand in culture. When murine ES cell-derived endothelial cells (MESDECs) were cultured on Matrigel and incubated for 48 h, vessel-like tube structures consisting of CD31 (PECAM-1) or BS-1 immunoreactive cells were developed. Immunocytochemistry and RT-PCR analyses revealed that MESDECs express endothelial cell-specific marker proteins such as Flk-1, PECAM-1, Tie-1, and Tie-2, in which the expressions persist for long periods of time after differentiation. The cells were also capable of taking up acetylated low-density lipoprotein (LDL) in culture. Our data suggest that MESDECs could provide a suitable in vitro model to study molecular events involved in vascular development and open up a new therapeutic strategy in regeneration medicine of cardiovascular disorders. PMID- 18393657 TI - Metallothionein downregulation in very old age: a phenomenon associated with cellular senescence? AB - It is known that metallothionein (MT) mRNA expression first increases with age, but then decreases again in the very elderly. Here we report that MT protein levels also decrease in very old age, and that this is independent of dietary zinc intake. Age-related changes of MT, as well as alterations of zinc homeostasis (intracellular labile zinc and NO-induced zinc release), occur both in human PBMCs ex vivo and also in CD4+ T cell clones progressing through their finite life span in vitro. These results suggest that phenomena observed in very old people can be at least partially attributed to diminished cell proliferation. PMID- 18393659 TI - Sugar absorption in the intestine: the role of GLUT2. AB - Intestinal glucose absorption comprises two components. One is classical active absorption mediated by the Na+/glucose cotransporter. The other is a diffusive component, formerly attributed to paracellular flow. Recent evidence, however, indicates that the diffusive component is mediated by the transient insertion of glucose transporter type 2 (GLUT2) into the apical membrane. This apical GLUT2 pathway of intestinal sugar absorption is present in species from insect to human, providing a major route at high sugar concentrations. The pathway is regulated by rapid trafficking of GLUT2 to the apical membrane induced by glucose during assimilation of a meal. Apical GLUT2 is therefore a target for multiple short-term and long-term nutrient-sensing mechanisms. These include regulation by a newly recognized pathway of calcium absorption through the nonclassical neuroendocrine l-type channel Cav1.3 operating during digestion, activation of intestinal sweet taste receptors by natural sugars and artificial sweeteners, paracrine and endocrine hormones, especially insulin and GLP-2, and stress. Permanent apical GLUT2, resulting in increased sugar absorption, is a characteristic of experimental diabetes and of insulin-resistant states induced by fructose and fat. The nutritional consequences of apical and basolateral GLUT2 regulation are discussed in the context of Western diet, processed foods containing artificial sweeteners, obesity, and diabetes. PMID- 18393660 TI - Protein and amino acid metabolism in the human newborn. AB - Birth and adaptation to extrauterine life involve major shifts in the protein and energy metabolism of the human newborn. These include a shift from a state of continuous supply of nutrients including amino acids from the mother to cyclic periodic oral intake, a change in the redox state of organs, thermogenesis, and a significant change in the mobilization and use of oxidative substrates. The development of safe, stable isotopic tracer methods has allowed the study of protein and amino acid metabolism not only in the healthy newborn but also in those born prematurely and of low birth weight. These studies have identified the unique and quantitative aspects of amino acid/protein metabolism in the neonate, thus contributing to rational nutritional care of these babies. The present review summarizes the contemporary data on some of the significant developments in essential and dispensable amino acids and their relationship to overall protein metabolism. Specifically, the recent data of kinetics of leucine, phenylalanine, glutamine, sulfur amino acid, and threonine and their relation to whole-body protein turnover are presented. Finally, the physiological rationale and the impact of nutrient (amino acids) interventions on the dynamics of protein metabolism are discussed. PMID- 18393661 TI - Role of nanomedicines in cell-based therapeutics. PMID- 18393663 TI - A new drug-sensing paradigm based on ion-current rectification in a conically shaped nanopore. AB - AIMS: To utilize the ion-current rectification phenomenon observed for conically shaped nanopores as the basis for designing sensors for drug molecules that adsorb to the walls of the nanopore. METHODS: The conically shaped nanopore was prepared by the well-known track-etch method in a polyimide (Kapton) membrane. The ion current flowing through the nanopore was measured as a function of applied transmembrane potential in the presence of the analyte drug molecule, Hoechst 33258. RESULTS: The pore walls in the Kapton membrane are hydrophobic yet have fixed carboxylate groups that give the walls a net negative charge. This fixed anionic surface charge causes the nanopore to rectify the ion current flowing through it. The analyte drug molecule, Hoechst 33258, is cationic yet also hydrophobic. When the membrane is exposed to this molecule, it adsorbs to the pore walls and neutralizes the anionic surface charge, thus lowering the extent of ion-current rectification. The change in rectification is proportional to the concentration of the drug. CONCLUSIONS: This nanopore sensor is selective for hydrophobic cations relative to anions, neutral molecules and less hydrophobic cations. Future work will explore ways of augmenting this hydrophobic effect-based selectivity so that more highly selective sensors can be obtained. PMID- 18393664 TI - Imaging characteristics of zinc sulfide shell, cadmium telluride core quantum dots. AB - AIMS: Quantum dots are optical nanocrystals whose in vitro and in vivo use in molecular imaging is expanding rapidly. In comparison with organic fluorophores, quantum dots exhibit desirable properties, such as multiwavelength fluorescence emission, excellent brightness and resistance to photobleaching. Their electron dense, metallic cores suggest utility in other clinical imaging modalities. METHODS: Core-shell zinc sulfide-cadmium telluride quantum dots were studied by magnetic resonance and computed tomography phantoms. Quantum dots were also injected into rat brain, as well as intravenously, using convection-enhanced delivery, prior to animal imaging. RESULTS: Computed tomography studies suggest that current formulations of quantum dots might be imaged in vivo in animals. CONCLUSIONS: Used in conjunction with optical imaging techniques, quantum dots have the potential to function as multimodal imaging platforms in vivo. The ability to detect an optical nanoparticle preoperatively with clinical imaging modality offers a distinct advantage to clinicians engaged in image-guided surgical applications. PMID- 18393665 TI - pH-triggered release of vancomycin from protein-capped porous silicon films. AB - OBJECTIVE: An in vitro model system for pH-triggered release of the antibiotic vancomycin from porous Si films is studied. METHOD: Vancomycin is infused into a mesoporous Si film from a mixed aqueous/acetonitrile solution and trapped by a capping layer containing the protein bovine serum albumin (BSA). The protein effectively traps vancomycin in the porous nanostructure at pH 4.0; the protein dissolves and vancomycin is released into solution when the pH increases to 7.4. The surface chemistry of porous Si exerts a substantial effect on the efficacy of drug loading. The amount of drug loading is larger in freshly-etched (hydrophobic, hydrogen-terminated) porous Si and smaller in methyl-modified, undecylenic acid-modified and thermally oxidized samples. The quantity of drug loaded in a freshly etched porous Si chip is proportional to the thickness of the porous layer, which exhibits a constant volume loading efficiency of 31% (v/v). Flow-cell experiments designed to mimic the transition from pH 4 to 7 that occurs when material moves from the stomach to the upper intestinal tract were performed on the freshly etched films and vancomycin- and BSA-release rates were quantified from the effluent of the flow cell by high-pressure liquid chromatography analysis. RESULTS & CONCLUSION: There is a small, constant rate of vancomycin release at pH 4 that is independent of the amount of drug loaded in the pores. This is attributed to diffusion of vancomycin from the BSA-capping layer. The release rate increases five- to tenfold when the pH of the solution in the flow cell increases to 7.4; 100% of the drug is released within 3 h of this increase. PMID- 18393666 TI - Electrospun biodegradable nanofibrous mats for tissue engineering. AB - AIMS & METHOD: In this study, a microbial polyester, poly(3-hydroxybutyrate-co- 3 hydroxyvalerate) (PHBV), and its blends were electrospun into PHBV (10% w/v), PHBV (15% w/v), PHBV-PLLA (5% w/v), PHBV-PLGA (50:50) (15% w/v) and PHBV P(L,DL)LA (5% w/v) fibrous scaffolds for tissue engineering. RESULTS: Various processing parameters affected the morphology and the dimensions of beads formed on the fibers. Concentration was highly influential on fiber properties; as it increased from 5 to 15% (w/v), fiber diameter increased from 284 +/- 133 nm to 2200 +/- 716 nm. Increase in potential (from 20 to 50 kV) did not lead to the expected decrease in fiber diameter. The blends of PHBV with lactide-based polymers led to fibers with less beads and more uniform diameter. The surface porosities for PHBV10, PHBV15, PHBV-PLLA, PHBV-PLGA (50:50) and PHBV-P(L,DL)LA were 38.0 +/- 3.8, 40.1 +/- 8.5, 53.8 +/- 4.2, 50.0 +/- 4.2 and 30.8 +/- 2.7%, respectively. In vitro studies using human osteosarcoma cells (Saos-2) revealed that the electrospun scaffolds promoted cell growth and penetration. Surface modification with oxygen plasma treatment slightly improved the improved the results in terms of cell number increase and significantly improved spreading of the cells. CONCLUSION: All scaffolds prepared by electrospinning have implied significant potential for use in further studies leading to bone tissue engineering applications. The PHBV-PLLA blend appeared to yield the best results regarding cell number increase, their attachment and spreading inside and on the scaffold. PMID- 18393667 TI - Osteoblastic cell behavior on nanostructured metal implants. AB - AIMS: Surface modifications at the nanometric scale may promote protein adsorption, cell adhesion and thus favor the osseointegration of metal implants. The behavior of osteoblastic cells was studied on mirror-polished (Smooth-SS) and nanostructured (Nano-SS) stainless steel surfaces. MATERIALS & METHODS: The nanostructuration was obtained by anodization. The surfaces were characterized by scanning electron, atomic force microscopy (AFM) and X-ray photoelectron specytoscopy. Osteoblastic cells were cultured on these surfaces. RESULTS & DISCUSSION: AFM showed a regular array of pores on the Nano-SS. Osteoblastic cells spread more rapidly on Nano-SS surfaces than on Smooth-SS surfaces. Cell viability was similar on Smooth-SS and Nano-SS. Alkaline phosphatase activity increased with culture time whatever the substrate but was better enhanced at 21 days on the Nano-SS than on the Smooth-SS. CONCLUSION: This study shows that the nanostructuration of metal implants may improve the adhesion and differentiation of osteoblastic cells. PMID- 18393668 TI - Quantum dots for cancer diagnosis and therapy: biological and clinical perspectives. AB - Quantum dots (QDs) are semiconductor nanocrystals that emit fluorescence on excitation with a light source. They have excellent optical properties, including high brightness, resistance to photobleaching and tunable wavelength. Recent developments in surface modification of QDs enable their potential application in cancer imaging. QDs with near-infrared emission could be applied to sentinel lymph-node mapping to aid biopsy and surgery. Conjugation of QDs with biomolecules, including peptides and antibodies, could be used to target tumors in vivo. In this review, we summarize recent progress in developing QDs for cancer diagnosis and treatment from a clinical standpoint and discuss future prospects of further improving QD technology to identify metastatic cancer cells, quantitatively measure the level of specific molecular targets and guide targeted cancer therapy by providing biodynamic markers for target inhibition. PMID- 18393670 TI - Biological and biomimetic molecular machines. AB - The evolution of life facilitates the creation of biological molecular machines. In these so-called 'nanomachines,' nature elegantly shows that when precisely organized and assembled, simple molecular mechanical components can link motions efficiently from the nanometer scale to the macroscopic world, and achieve complex functions such as powering skeletal muscles, synthesizing ATP and producing DNA/RNA. Inspired by nature, researchers are creating artifical molecular machines with tailored structures and properties, with the aim of realizing man-made active nanosystems that operate with the same efficiency and complexity as biological nanomachines. It is anticipated that in the not-too distant future, unique applications of biological and biomimetic molecular machines will emerge in areas such as biochemical instrumentation and nanomedicine. PMID- 18393671 TI - Electron tomography in nanoparticle imaging and analysis. AB - A wide range of medically important nanosized biological assemblies are not amenable to study by standard structural techniques, such as x-ray crystallography or nuclear magnetic resonance spectroscopy, either owing to their large size or the intrinsic heterogeneity of the specimen. The emerging technique of cryo-electron tomography is being applied actively to study these nanoparticles and has the potential of providing high-resolution structural information on these heterogeneous assemblies. Although the majority of structural methods involve the averaging of large numbers of structurally homogeneous molecules, tomography enables the visualization and quantitation of variation in a mixed population. Here, we present a review of the principles of cryo-electron tomography as applied to the 3D analysis of nanoparticles and illustrate applications where it can be used for visualizing the architecture of enveloped viruses and for the analysis of size and compositional variation of Doxil, a commonly used, US FDA-approved nanomedicine. PMID- 18393669 TI - Understanding and re-engineering nucleoprotein machines to cure human disease. AB - The mammalian nucleus is filled with self-organizing, nanometer-scale nucleoprotein machines that carry out DNA replication, RNA biogenesis and DNA repair. We discuss, as a model, the nonhomologous end-joining (NHEJ) machine, which repairs DNA double-strand breaks. The NHEJ machine consists of six core polypeptides and 10-20 ancillary polypeptides. A full understanding of its design principles will require measuring the behavior of single NHEJ complexes in living cells, using a Nano Toolbox that includes bright, stable, biocompatible fluorophores, efficient protein and nucleic acid-tagging strategies, and sensitive, high-resolution imaging methods. Taking inspiration from natural examples, it might be possible to adapt and redesign the NHEJ machine to precisely correct mutations responsible for common human diseases, such as K-ras in lung cancer or human papillomavirus E6 and E7 genes in cervical and oral cancers. PMID- 18393673 TI - Teratoma formation by human embryonic stem cells is site dependent and enhanced by the presence of Matrigel. AB - When implanted into immunodeficient mice, human embryonic stem cells (hESCs) give rise to teratoma, tumor-like formations containing tissues belonging to all three germ layers. The ability to form teratoma is a sine qua non characteristic of pluripotent stem cells. However, limited data are available regarding the effects of implantation site and the methods employed for implantation on the success rate of teratoma formation. In this study, the rate of teratoma formation in immunodeficient mice was site dependent: subcutaneous (25-100%), intratesticular (60%), intramuscular (12.5%), and under the kidney capsule (100%). Co-injecting the hESCs with Matrigel increased subcutaneous teratoma formation efficiency from 25-40% to 80-100%. We did not observe site-specific differences in the teratoma composition at the histological level. However, subcutaneous teratomas were quite distinct, easy to remove, and caused minimal discomfort to the mice. Also, subcutaneous teratomas displayed larger proportion of solid tissues as opposed to cyst formation that dominated the teratomas formed at the other sites. Interestingly, a chromosomally abnormal hESCs with trisomy 20 formed teratomas where the ratio of differentiated to undifferentiated tissues was significantly decreased suggesting defective pluripotency of the cells. In conclusion, subcutaneous implantation of hESCs in presence of Matrigel appears to be the most efficient, reproducible, and the easiest approach for teratoma formation by hESCs. Also, teratoma formation can be employed to study the development defects exhibited by the chromosomally abnormal hESC lines. PMID- 18393672 TI - Detection of transketolase in bone marrow-derived insulin-producing cells: benfotiamine enhances insulin synthesis and glucose metabolism. AB - Adult bone marrow (BM)-derived insulin-producing cells (IPCs) are capable of regulating blood glucose levels in chemically induced hyperglycemic mice. Using cell transplantation therapy, fully functional BM-derived IPCs help to mediate treatment of diabetes mellitus. Here, we demonstrate the detection of the pentose phosphate pathway enzyme, transketolase (TK), in BM-derived IPCs cultured under high-glucose conditions. Benfotiamine, a known activator of TK, was not shown to affect the proliferation of insulinoma cell line, INS-1; however, when INS-1 cells were cultured with oxythiamine, an inhibitor of TK, cell proliferation was suppressed. Treatment with benfotiamine activated glucose metabolism in INS-1 cells in high-glucose culture conditions, and appeared to maximize the BM-derived IPCs ability to synthesize insulin. Benfotiamine was not shown to induce the glucose receptor Glut-2, however it was shown to activate glucokinase, the enzyme responsible for conversion of glucose to glucose-6-phosphate. Furthermore, benfotiamine-treated groups showed upregulation of the downstream glycolytic enzyme, glyceraldehyde phosphate dehydrogenase (GAPDH). However, in cells where the pentose phosphate pathway was blocked by oxythiamine treatment, there was a clear downregulation of Glut-2, glucokinase, insulin, and GAPDH. When benfotiamine was used to treat mice transplanted with BM-derived IPCs transplanted, their glucose level was brought to a normal range. The glucose challenge of normal mice treated with benfotiamine lead to rapidly normalized blood glucose levels. These results indicate that benfotiamine activates glucose metabolism and insulin synthesis to prevent glucose toxicity caused by high concentrations of blood glucose in diabetes mellitus. PMID- 18393674 TI - From the determination of complex reaction mechanisms to systems biology. AB - This review presents several methods of determining complex chemical reaction mechanisms and their functions. One method is based on correlation functions of measured time series of concentrations of chemical species, another is on measurements of temporal responses of concentrations to various perturbations of arbitrary magnitude, the third deals with the analysis of oscillatory systems, and the fourth describes the use of genetic algorithms. All methods are applicable to chemical, biochemical, and biological reaction systems and to genetic networks. The methods depend on the design of appropriate experiments for the whole system and corresponding theories for interpretation that lead to information on the causal chemical connectivity of species, reaction pathways, reaction mechanisms, control centers in the system, and functions of the system. The first three methods require no assumption of a model or hypothesis, nor extensive calculations, unlike the interpretation of measurements made on a gene network at only one time. The methods offer advantageous approaches to systems biology. PMID- 18393675 TI - Chemistry and photochemistry of mineral dust aerosol. AB - It has become increasingly clear that heterogeneous and multiphase chemistry of tropospheric aerosols can change the chemical balance of the atmosphere. In this review, we focus on recent laboratory studies of the heterogeneous and multiphase chemistry and photochemistry of mineral dust aerosol, a large mass fraction of the tropospheric aerosol. Mineral dust aerosol contains a mixture of oxides, clays, and carbonates. Molecular-based studies of reactions of these dust components provide insights into the chemistry of Earth's atmosphere. We discuss several different types of heterogeneous and multiphase reactions, including (a) ozone decomposition, (b) nitrogen dioxide and nitrate photochemistry, and (c) the dissolution and redox chemistry of Fe-containing dust. We also review some of the important chemical concepts that have recently emerged. PMID- 18393676 TI - Energy flow in proteins. AB - Energy flows anisotropically through the residues and vibrational states of globular proteins. A variety of experimental and computational studies have identified energy transport channels traversing many residues, in some cases connecting functional regions, potentially important in allostery, and in other cases having no apparent function. This property and the diffusion of energy in proteins are mimicked by transport on a percolation cluster. I review work that addresses connections between globular proteins, percolation clusters, and the similarity of energy flow and thermal transport in these systems. I also review experimental and theoretical studies of the anisotropic flow of energy through the vibrational states of a protein, a property that also can be understood by comparison with simple model disordered systems. PMID- 18393677 TI - Functional polymer brushes in aqueous media from self-assembled and surface initiated polymers. AB - This review focuses on the behavior of single-component, water-soluble neutral and charged brushes. Selected examples illustrate how solvation effects, hydrophobic interactions, and electrostatic interactions create complex behaviors not easily captured in mean-field treatments. In particular, we distinguish between two classes of polymer brushes: those that can be described classically within the context of generalized van der Waals potentials and those that can be described by model-dependent potentials arising from specific interactions. In classical systems, only a few global parameters are needed to predict behavior. Nonclassical systems, in contrast, necessitate several local details, which do not necessarily lead to universal scaling laws. Although these nonclassical interactions present unique opportunities for engineering functional surfaces, they also present new challenges for designing well-defined systems with precise control over distributions in the degree of polymerization and tethering density. PMID- 18393678 TI - Electronic spectroscopy of carbon chains. AB - Investigators have recorded the electronic spectra of assorted carbon-chain systems in the gas phase using a variety of methods, ranging from direct cavity ringdown absorption spectroscopy to photofragmentation techniques that utilize the cooling capabilities of an ion trap. We summarize the results from these studies and compare them with astronomical measurements of the diffuse interstellar band (DIB) absorptions. Although carbon chains comprising up to a handful of carbon atoms cannot be the carrier species, we explore which chains remain viable. In particular, the (1)Sigma(u)(+)-Chi(1)Sigma(g)(+) transitions of the odd-numbered carbon chains (n = 17,19,...) possess large oscillator strengths and lie in the 400-900-nm DIB range. The origin bands of larger bare carbon rings, such as C(18), have also been observed, with striking similarities to some DIB measurements at high resolution, although at other wavelengths. Finally, we consider recently obtained electronic spectra of metal-containing carbon chains. PMID- 18393680 TI - Fluctuation theorems. AB - Fluctuation theorems, developed over the past 15 years, have resulted in fundamental breakthroughs in our understanding of how irreversibility emerges from reversible dynamics and have provided new statistical mechanical relationships for free-energy changes. They describe the statistical fluctuations in time-averaged properties of many-particle systems such as fluids driven to nonequilibrium states and provide some of the few analytical expressions that describe nonequilibrium states. Quantitative predictions on fluctuations in small systems that are monitored over short periods can also be made, and therefore the fluctuation theorems allow thermodynamic concepts to be extended to apply to finite systems. For this reason, we anticipate an important role for fluctuation theorems in the design of nanotechnological devices and in the understanding of biological processes. This review discusses these theorems, their physical significance, and results for experimental and model systems. PMID- 18393679 TI - Free energies of chemical reactions in solution and in enzymes with ab initio quantum mechanics/molecular mechanics methods. AB - Combined quantum mechanics/molecular mechanics (QM/MM) methods provide an accurate and efficient energetic description of complex chemical and biological systems, leading to significant advances in the understanding of chemical reactions in solution and in enzymes. Here we review progress in QM/MM methodology and applications, focusing on ab initio QM-based approaches. Ab initio QM/MM methods capitalize on the accuracy and reliability of the associated quantum-mechanical approaches, however, at a much higher computational cost compared with semiempirical quantum-mechanical approaches. Thus reaction-path and activation free-energy calculations based on ab initio QM/MM methods encounter unique challenges in simulation timescales and phase-space sampling. This review features recent developments overcoming these challenges and enabling accurate free-energy determination for reaction processes in solution and in enzymes, along with applications. PMID- 18393681 TI - Structure, dynamics, and assembly of filamentous bacteriophages by nuclear magnetic resonance spectroscopy. AB - Filamentous bacteriophages serve as model systems for the development and implementation of spectroscopic methods suitable for biological supramolecular assemblies. Not only are their coat proteins small and readily prepared in the laboratory, but they also have two primary roles as membrane proteins and as the principal structural element of the virus particles. As a bacterial system, they are readily labeled with stable isotopes, and this has opened possibilities for the many nuclear magnetic resonance (NMR) studies described in this review. In particular, solid-state NMR of aligned samples has been used to determine the three-dimensional structures of both the membrane-bound forms of coat proteins in phospholipid bilayers and structural forms in virus particles, which has led to an analysis of the assembly mechanism for virus particles as they are extruded through the cell membrane. PMID- 18393682 TI - Inside a collapsing bubble: sonoluminescence and the conditions during cavitation. AB - Acoustic cavitation, the growth and rapid collapse of bubbles in a liquid irradiated with ultrasound, is a unique source of energy for driving chemical reactions with sound, a process known as sonochemistry. Another consequence of acoustic cavitation is the emission of light [sonoluminescence (SL)]. Spectroscopic analyses of SL from single bubbles as well as a cloud of bubbles have revealed line and band emission, as well as an underlying continuum arising from a plasma. Application of spectrometric methods of pyrometry as well as tools of plasma diagnostics to relative line intensities, profiles, and peak positions have allowed the determination of intracavity temperatures and pressures. These studies have shown that extraordinary conditions (temperatures up to 20,000 K; pressures of several thousand bar; and heating and cooling rates of >10(12) K s(1)) are generated within an otherwise cold liquid. PMID- 18393683 TI - Collapse mechanisms of Langmuir monolayers. AB - When a two-dimensional (2D) film is compressed to its stability limit, it explores the third dimension via collapse. Understanding this 2D-to-3D transition is of great importance as it provides insight into the origin of defects in thin films. This review draws attention to a reversible folding collapse first discovered in model lung surfactant systems and explores the driving forces for this mechanism. The mode of collapse can be tuned by varying the mechanical properties of the film. I present a continuum elastic theory that captures the onset of the observed folding instability and use digital image analysis to analyze the folding dynamics. This article further explores factors that determine the maximum surface pressure a mixed monolayer can sustain and explains the observed phenomenon using the principle of rigidity percolation. The folding transition observed in lipid monolayers described here has also been observed in other systems, including monolayers of nanoparticles. PMID- 18393684 TI - Human cord blood mononuclear cells decrease cytokines and inflammatory cells in acute myocardial infarction. AB - We investigated whether human umbilical cord blood mononuclear cells (HUCBC), which contain hematopoietic and mesenchymal progenitor cells, can limit myocardial cytokine expression and inflammatory cell infiltration in acute myocardial infarction. We permanently ligated the left coronary artery of rats and injected into the myocardium either Isolyte or 4 x 10(6) HUCBC in Isolyte and measured myocardial cytokines with antibody arrays at 2, 6, 12, 24, and 72 hours after infarction. We then measured with flow cytometry myocardial macrophages, neutrophils and lymphocytes at 12, 24, and 72 hours after infarctions in rats treated with either intramyocardial Isolyte or 4 x 10(6) HUCBC. In the Isolyte treated hearts, between 2 and 12 hours after myocardial infarction, tumor necrosis factor-alpha increased from 6.7 +/- 0.9% to 52.3 +/- 4.7%, monocyte chemoattract protein increased from 9.5 +/- 1.2% to 39.8 +/- 2.1%, fractalkine increased from 11 +/- 1.5% to 28.1 +/- 1.3%, ciliary neurotrophic factor increased from 12.1 +/- 0.02% to 25.9 +/- 1.1%, macrophage inflammatory protein increased from 10.3 +/- 1.5% to 23.9.0 +/- 1.4%, interferon-gamma increased from 8.7 +/- 0.4% to 26.0 +/- 1.6%, interleukin-1beta increased from 6.1 +/- 0.04% to 19.0 +/- 1.2%, and IL-4 increased from 5.9 +/- 0.03% to 15 +/- 1.5% (all p < 0.001 compared with controls). The concentrations of fractalkine remained significantly increased at 72 hours after acute infarction. In contrast, the myocardial concentrations of these cytokines did not significantly change in HUCBC treated hearts at 2, 6, 12, 24, or 72 hours after infarction. The percentage of neutrophils increased from 0.04 +/- 0.2%/50,000 heart cells in the controls to 5.3 +/- 1.2%/50,000 heart cells 12 hours after infarction in Isolyte treated hearts but averaged only 1.3 +/- 0.7%/50,000 heart cells in HUCBC treated hearts (p < 0.02). Thereafter, the percentages of neutrophils rapidly decreased at 24 and at 72 hours after infarction and averaged 0.6 +/- 0.2%/50,000 heart cells at 72 hours after infarction in Isolyte-treated hearts in contrast to 0.2 +/- 0.1%/50,000 cells in HUCBC hearts (p < 0.05). Moreover, the percentages of neutrophils at 24 and 72 hours in HUCBC hearts were not significantly different from controls. At 24 hours post infarction, the percentage of CD3 and CD4 lymphocytes were 10.7 +/- 1.4% and 6.3 +/- 1.1%/50,000 cells in Isolyte hearts in comparison with only 4.9 +/- 0.8% and 2.9 +/- 0.5% in HUCBC hearts (p < 0.005 for Isolyte versus HUCBC). The percentage of CD11b macrophages was 2.8 +/- 0.3% in Isolyte hearts and 1.9 +/- 0.2% in HUCBC treated hearts (p < 0.05). At 72 hours after infarction, the percentage of CD3 and CD4 lymphocytes averaged 8.0 +/- 1.1% and 5.1 +/- 0.8%/50,000 heart cells in Isolyte hearts in comparison with only 4.1 +/- 0.5% and 2.3 +/- 0.4%/50,000 heart cells in the HUCBC treated infarctions (p < 0.005). Left ventricular infarct sizes in Isolyte-treated hearts at 72 hours post infarction averaged 15.7 +/- 1.4% of the left ventricular muscle area in contrast to HUCBC treated infarctions that averaged 6.9 +/- 1.4% of the left ventricular muscle area (p < 0.02). Moreover in rats followed for 2 months post infarction, the LV ejection fractions decreased to 65.4 +/- 1.9% and 69.1 +/- 1.9% at 1 and 2 months after infarction in Isolyte-treated hearts and were significantly different from HUCBC treated hearts that averaged 72.1 +/- 1.3% and 75.7 +/- 1.4% (both p < 0.02). The present experiments suggest that an important mechanism whereby HUCBC limit infarct size and improve left ventricular ejection fraction is by significantly limiting inflammatory cytokines and inflammatory cells in infarcted myocardium. PMID- 18393685 TI - Movement of fumigants in soil, dosage responses, and differential effects. PMID- 18393686 TI - Rydberg molecules. PMID- 18393687 TI - Prevalence and clinical relevance of occult hepatitis B in the fibrosis progression and antiviral response to INF therapy in HIV-HCV-coinfected patients. AB - Occult hepatitis B virus (HBV) infection is diagnosed when HBc antibodies (HBcAb) and HBV DNA are detectable in serum while hepatitis B surface antigen (HBsAg) is not. This situation has been frequently described in patients with chronic hepatitis C virus (HCV) infection. The objective of this study was to evaluate the prevalence of occult hepatitis B in HIV-HCV-coinfected patients and its clinical relevance in liver histology and viral response after interferon therapy for HCV. A total of 238 HIV-HCV-infected patients,negative for HBsAg, were included. Serum samples were analyzed for the presence of HBV DNA and HBcAb.HBV DNA quantification was determined with the Cobas TaqMan HBV Test (detection limit 6 IU/ml). Data from liver biopsy and laboratory tests were also analyzed. HBcAb resulted in 142 (60%) patients, being the independent associated factors: male gender, previous history of intravenous drug use, age, CD4 count,and HAV antibody presence. Among 90 HBcAb patients that we could analyze, HBV DNA was positive in 15 (16.7% of occult hepatitis B infection in this group, and 6.3% in the whole HIV-HCV cohort studied). No baseline factors, liver histology, or HCV therapy response were related to the presence of HBV DNA. We found that occult hepatitis B is a frequent condition present in at least 6.3% of our HCV-HIV patients and in more than 16% of those with HBcAb. Despite the high prevalence, this phenomenon does not seem to affect the clinical evolution of chronic hepatitis C or modify the viral response to interferon-based HCV therapies PMID- 18393688 TI - Testing the H2O2-H2O hypothesis for life on Mars with the TEGA instrument on the Phoenix lander. AB - In the time since the Viking life-detection experiments were conducted on Mars, many missions have enhanced our knowledge about the environmental conditions on the Red Planet. However, the martian surface chemistry and the Viking lander results remain puzzling. Nonbiological explanations that favor a strong inorganic oxidant are currently favored (e.g., Mancinelli, 1989; Plumb et al., 1989; Quinn and Zent, 1999; Klein, 1999; Yen et al., 2000), but problems remain regarding the lifetime, source, and abundance of that oxidant to account for the Viking observations (Zent and McKay, 1994). Alternatively, a hypothesis that favors the biological origin of a strong oxidizer has recently been advanced (Houtkooper and Schulze-Makuch, 2007). Here, we report on laboratory experiments that simulate the experiments to be conducted by the Thermal and Evolved Gas Analyzer (TEGA) instrument of the Phoenix lander, which is to descend on Mars in May 2008. Our experiments provide a baseline for an unbiased test for chemical versus biological responses, which can be applied at the time the Phoenix lander transmits its first results from the martian surface. PMID- 18393689 TI - Analysis of underivatized amino acids in geological samples using ion-pairing liquid chromatography and electrospray tandem mass spectrometry. AB - The capability of detecting biomarkers, such as amino acids, in chemically complex field samples is essential to establishing the knowledge required to search for chemical signatures of life in future planetary explorations. However, due to the complexities of in situ investigations, it is important to establish a new analytical scheme that utilizes a minimal amount of sample preparation. This paper reports the feasibility of a novel and sensitive technique, which has been established to quantitate amino acids in terrestrial crust samples directly without derivatization using volatile ion-pairing liquid chromatography and tandem mass spectrometry equipped with an electrospray ionization source. Adequate separation of 20 underivatized amino acids was achieved on a C(18) capillary column within 26 min with nonafluoropentanoic acid (NFPA) as ion pairing reagent. Each amino acid was identified from its retention time as well as from its characteristic parent-to-daughter ion transition. Using tandem mass spectrometry as a detection technique allows co-elution of some amino acids, as it is more specific than traditional spectrophotometric methods. In the present study, terrestrial samples collected from 3 different locations were analyzed for their water-extractable free amino acid contents, following the removal of metal and organic interferences via ion exchange procedures. This is the first time that amino acids in geological samples were directly determined quantitatively without complicated derivatization steps. Depending on the amino acid, the detection limits varied from 0.02 to 5.7 pmol with the use of a 1 microl sample injection loop. PMID- 18393690 TI - On the survivability and detectability of terrestrial meteorites on the moon. AB - Materials blasted into space from the surface of early Earth may preserve a unique record of our planet's early surface environment. Armstrong et al. (2002) pointed out that such materials, in the form of terrestrial meteorites, may exist on the Moon and be of considerable astrobiological interest if biomarkers from early Earth are preserved within them. Here, we report results obtained via the AUTODYN hydrocode to calculate the peak pressures within terrestrial meteorites on the lunar surface to assess their likelihood of surviving the impact. Our results confirm the order-of-magnitude estimates of Armstrong et al. (2002) that substantial survivability is to be expected, especially in the case of relatively low velocity (ca. 2.5 km/s) or oblique (amino acids-->oligomers. The simulations were run for the formation of all oligomers up to the 20-mer over a 1 Gy interval from the end of the period of heavy bombardment, during which period life emerged. Upper-limit rate constant estimates developed from the studies of Huber and Wachtershauser were employed. The simulations showed that oligomer production consistent with life's start within that interval emerges only with an autocatalyst exhibiting a catalytic proficiency comparable to that of contemporary enzymes. The simulations, moreover, ignored likely thermodynamic and statistical burdens which, if included, would have led to the need for catalytic capacities well in excess of those in present-day enzymes. Prebiotic oligomers with such levels of activity are clearly not likely, and it is apparent that the iron-sulfur scheme could not have played a role in life's beginnings. PMID- 18393693 TI - Rate measurements of the hydrolysis of complex organic macromolecules in cold aqueous solutions: implications for prebiotic chemistry on the early Earth and Titan. AB - Organic macromolecules ("complex tholins") were synthesized from a 0.95 N(2)/0.05 CH(4) atmosphere in a high-voltage AC flow discharge reactor. When placed in liquid water, specific water soluble compounds in the macromolecules demonstrated Arrhenius type first order kinetics between 273 and 313 K and produced oxygenated organic species with activation energies in the range of approximately 60+/-10 kJ mol(-1). These reactions displayed half lives between 0.3 and 17 days at 273 K. Oxygen incorporation into such materials--a necessary step toward the formation of biological molecules--is therefore fast compared to processes that occur on geologic timescales, which include the freezing of impact melt pools and possible cryovolcanic sites on Saturn's organic-rich moon Titan. PMID- 18393736 TI - Can liability rules keep pace with best practice? The case of multidisciplinary cancer care. PMID- 18393737 TI - Will prasugrel supersede clopidogrel for acute coronary syndromes? PMID- 18393738 TI - New vision, new paradigm: health and wealth for all by 2100--or for no one. PMID- 18393739 TI - National Breast Cancer Audit: the use of multidisciplinary care teams by breast surgeons in Australia and New Zealand. AB - OBJECTIVE: To explore the involvement of members of the Royal Australasian College of Surgeons (RACS) Section of Breast Surgery in Australia and New Zealand in multidisciplinary care (MDC) teams. DESIGN AND SETTING: Questionnaire sent to all full members of the RACS Section of Breast Surgery in December 2006. PARTICIPANTS: 239 of 262 active full members of the RACS Section of Breast Surgery (response rate, 91.2%). MAIN OUTCOME MEASURES: Surgeons' use of, and the composition and functioning of, MDC teams in public and private practice, and in metropolitan, regional and rural settings. RESULTS: 85% of responding surgeons reported participating in at least one fully established MDC team. Public-sector teams were operationally more consistent and functional than private teams, and rural teams were less well developed than those in metropolitan and regional centres. The six core disciplines recommended by the National Breast Cancer Centre appear to be well represented in most teams. Patients and their general practitioners were not considered to be part of the treatment team by surgeons. CONCLUSIONS: MDC is supported by most breast surgeons, but there are deficits in rural areas, and in the private sector relative to the public sector. PMID- 18393740 TI - When continuity of care breaks down: a systems failure in identification of osteoporosis risk in older patients treated for minimal trauma fractures. AB - OBJECTIVE: Minimal trauma fractures may be the first indication of osteoporosis. Our aim was to determine the proportion of patients who underwent bone density testing for osteoporosis of those with a minimal trauma wrist fracture treated in the emergency department (ED). DESIGN: This observational retrospective cohort study used explicit medical record review and scripted telephone interviews. SETTING: EDs of three metropolitan hospitals in Melbourne in 2006. PARTICIPANTS: Patients aged 50 years and over who were treated for wrist fracture due to minimal trauma. Data collected included demographic details, fracture details, causes of injury, any bone density testing and any osteoporosis-related medication change. MAIN OUTCOME MEASURE: The proportion of patients who underwent bone density testing in the follow-up period. RESULTS: 131 patients were studied; 83% were female, and the median age was 71 years. No patient was referred by an ED or fracture clinic for bone density testing (95% CI, 0-3.5%). Telephone follow up data were obtained from 91 patients. Of these, 28 reported having bone density testing after their fracture, of whom 14 (50%; 95% CI, 32%-67%) were found to have osteoporosis. Seven were treated with a bisphosphonate and one with a selective oestrogen-receptor modulator. CONCLUSION: Follow-up of patients suffering minimal trauma wrist fractures treated in the ED is poor. Systems to improve the identification and treatment of osteoporosis in this group are needed if future osteoporotic fractures and their consequences are to be avoided. PMID- 18393741 TI - Teenage smoking in pregnancy and birthweight: a population study, 2001-2004. AB - OBJECTIVE: To determine the association between smoking in pregnant teenagers and baby birthweight. DESIGN, SETTING AND PARTICIPANTS: A retrospective population based study of women aged < 20 years who gave birth to liveborn singletons in Australia between January 2001 and December 2004. Data were drawn from the National Perinatal Data Collection. MAIN OUTCOME MEASURES: Maternal smoking, birthweight, low birthweight (LBW). RESULTS: The prevalence of LBW in babies born to teenage smokers was 9.9%, compared with 6.0% in babies born to teenage non smokers (odds ratio [OR], 1.72 [95% CI, 1.57-1.90]). On average, babies born to teenage smokers were 179.8 g lower in birthweight than babies born to teenage non smokers (95% CI, 165.5 -194.1 g; t = 24.6, P < 0.001). Smoking, Indigenous status, Socio-Economic Indexes for Areas category and parity were independently associated with LBW (all ORs > 1.3; P < 0.001) after adjusting for maternal age group. Teenagers smoking > 10 cigarettes a day had babies with lower birthweight that those who smoked < or = 10 cigarettes a day, demonstrating a dose-response relationship. The babies of teenage smokers who stopped smoking before 20 weeks' gestation had birthweights similar to those of babies born to teenage non smokers. One in 15 teenage smokers stopped smoking during pregnancy. CONCLUSION: Babies whose mothers smoked during pregnancy were more likely to have LBW than babies whose mothers did not smoke. Mothers who continue to smoke in the second half of pregnancy increase their baby's risk of LBW. There is significant scope to improve the quitting rate, and health professionals need to target smoking cessation at all contacts with pregnant women who continue to smoke. PMID- 18393742 TI - The National Open Disclosure Pilot: evaluation of a policy implementation initiative. AB - OBJECTIVE: To determine which aspects of open disclosure "work" for patients and health care staff, based on an evaluation of the National Open Disclosure Pilot. DESIGN, SETTING AND PARTICIPANTS: Qualitative analysis of semi-structured and open-ended interviews conducted between March and October 2007 with 131 clinical staff and 23 patients and family members who had participated in one or more open disclosure meetings. 21 of 40 pilot hospital sites, in New South Wales, South Australia, Victoria and Queensland, were included in the evaluation. Participating health care staff comprised 49 doctors, 20 nurses, and 62 managerial and support staff. In-depth qualitative data analysis involved mapping of discursive themes and subthemes across the interview transcripts. RESULTS: Interviewees broadly supported open disclosure; they expressed uncertainty about its deployment and consequences, and made detailed suggestions of ways to optimise the experience, including careful pre-planning, participation by senior medical staff, and attentiveness to consumers' experience of the adverse event. CONCLUSION: Despite some uncertainties, the national evaluation indicates strong support for open disclosure from both health care staff and consumers, as well as a need to resource this new practice. PMID- 18393743 TI - Medicolegal implications of a multidisciplinary approach to cancer care: consensus recommendations from a national workshop. AB - Concerns about medicolegal implications of a multidisciplinary approach to cancer care may act as a barrier to the implementation of best practice approaches. While multidisciplinary meetings carry a low level of medicolegal risk, improved documentation and transparency in approach will assist in limiting liability for individual health professionals and health services. The medicolegal implications of a multidisciplinary approach are not affected by whether a health professional bills the patient for attendance at multidisciplinary meetings. PMID- 18393744 TI - Implementing the national priorities for injury surveillance. AB - Injury is a leading cause of disability and death in Australia and is recognised as a national health priority area. The foundation of successful injury prevention is injury surveillance, and national policies and strategies developed over the past 20 years to reduce the burden of injury in Australia have included 22 recommendations on surveillance--only three of which have been completely implemented. Priorities for improving injury surveillance include: improving current injury mortality and morbidity data collection systems; filling the gaps in injury surveillance; maintaining vigilance over data quality; increasing the integration and accessibility of injury data; developing technical expertise in surveillance. Barriers to implementation of the current National Injury Prevention and Safety Promotion Plan include the lack of an implementation plan, performance management structure, appropriate national governance structure and resources--all of which could be overcome with government commitment. PMID- 18393745 TI - Management of adrenal insufficiency during the stress of medical illness and surgery. AB - Patients with adrenal insufficiency (AI) require additional glucocorticoid doses during surgery or medical illness, but there is no universally accepted regimen for glucocorticoid supplementation therapy. The high doses and long duration of glucocorticoid coverage that have traditionally been used do not reflect the hypothalamic-pituitary-adrenal response to surgical stress and medical illness in normal people. While the optimal dose and duration of supplementation therapy have not been established, our recommendations are based on extrapolation from what constitutes a normal cortisol response to stress, on expert opinion derived from the medical literature, and on clinical experience. The recommended use of lower doses of glucocorticoids during surgical and medical stress should not de emphasise the importance of additional supplementation during such events. Our recommendations do not replace clinical judgement, but their use will ensure that patients with AI are safely managed during illness or surgery without the risk of an adrenal crisis or excessive steroid dosing. PMID- 18393746 TI - Empathy and competence. AB - There has been a call to include empathy as a selection criterion in medical training. Empathy is a complex construct currently assessed by self-rating and observational scales, which may be complicated by the subjectivity of such measurements. Neuroscientific research into disorders of empathy such as autism should be encouraged to help further refine the evolving construct of empathy. Empathy may be more common in females, and selection for higher empathy may discriminate against males unless sex-specific adjustments are included in selection criteria. Physician empathy may lead to greater patient satisfaction and confidence in physicians, but more evidence is needed to support links to physician competence. In contrast, academic performance and conscientiousness have been clearly linked to physician competence. Competence and empathy may be independent qualities developed by different aspects of medical training. Provision of better work conditions and environments for physicians may forestall erosion of empathy, reducing the need to predict and enhance its development. Empathy should be valued in medical students and doctors, but more research is needed into the nature, assessment, and correlates of empathy before its adoption as a selection criterion for medical students. PMID- 18393747 TI - Haemopericardium in blue rubber bleb naevus syndrome (Bean syndrome). PMID- 18393749 TI - What has happened to clinical leadership in futile care discussions? PMID- 18393750 TI - Liver failure associated with the use of black cohosh for menopausal symptoms. PMID- 18393752 TI - Ready, SET, go for academic surgery? PMID- 18393753 TI - Fatal paramethoxy-amphetamine (PMA) poisoning in the Australian Capital Territory. PMID- 18393754 TI - Chronic kidney disease and automatic reporting of estimated glomerular filtration rate: revised recommendations. PMID- 18393755 TI - Successful lung transplantation for adolescents at a hospital for adults. PMID- 18393756 TI - Informing prospective medical students. PMID- 18393757 TI - Role of A-chain in functioning of the active site of human alpha-thrombin. AB - This review summarizes current data suggesting that A-chain of the human alpha thrombin molecule plays a role of allosteric effector in catalytic reactions with various substrates. Special attention is paid to the relationship between A-chain structure and catalytic activity of thrombin. The existence of this relationship is based on studies of natural mutation of A-chain of the alpha-thrombin molecule. Use of molecular and essential dynamics confirmed the role of A-chain in changes of conformation and catalytic properties of this enzyme; these changes involve residues located in the specificity sites and some inserting loops. Current knowledge on structure and properties of thrombin can be used for the development of new antithrombin agents. PMID- 18393758 TI - Synthesis, characterization, and studies on DNA binding of a new Mg(II) complex with N1,N8-bis(1-methyl-4-nitropyrrole-2-carbonyl)triethylenetetramine. AB - A new Mg(II) complex of MgL(NO3)2 (here L = N(1),N(8)-bis(1-methyl-4-nitropyrrole 2-carbonyl)triethylenetetramine) has been synthesized and characterized. The interactions between the Mg(II) complex and calf thymus DNA has been investigated using UV spectra, fluorescent spectra, viscosity, thermal denaturation, and molecular modeling. The cleavage reaction on plasmid DNA has been monitored by agarose gel electrophoresis. The experimental results show that the mode of binding of the complex to DNA is non-classical electrostatic action and the complex can cleave pBR322 DNA. PMID- 18393759 TI - Interaction between kringle and growth-factor-like domains in the urokinase molecule: possible role in stimulation of chemotaxis. AB - The results presented in this paper suggest the presence of an interaction between the kringle- and the growth-factor-like urokinase domains. This interaction regulates chemotactic properties of urokinase. We also show that interaction of urokinase with its "classical" receptor (uPAR) has a "permissive" effect on the interactions between the kringle domain and other targets on the cell surface. On the basis of our data we can suggest that uPAR serves as an "adaptor" for urokinase, and the binding of urokinase kringle domain to its receptor causes immediate activation of intracellular signaling and induction of cell migration. PMID- 18393760 TI - Interaction of APE1 and other repair proteins with DNA duplexes imitating intermediates of DNA repair and replication. AB - Interactions of APE1 (human apurinic/apyrimidinic endonuclease 1) and DNA polymerase beta with various DNA structures imitating intermediates of DNA repair and replication were investigated by gel retardation and photoaffinity labeling. Photoaffinity labeling of APE1 and DNA polymerase beta was accomplished by DNA containing photoreactive group at the 3 -end in mouse embryonic fibroblast (MEF) cell extract or for purified proteins. On the whole, modification efficiency was the same for MEF-extract proteins and for purified APE1 and DNA polymerase beta depending on the nature of the 5 -group of a nick/gap in the DNA substrate. Some of DNA duplexes used in this work can be considered as short-patch (DNA with the 5 -phosphate group in the nick/gap) or long-patch (DNA containing 5 -sugar phosphate or 5 -flap) base excision repair (BER) intermediates. Other DNA duplexes (3 -recessed DNA and DNA with the 5 -hydroxyl group in the nick/gap) have no relation to intermediates forming in the course of BER. As shown by both methods, APE1 binds with the highest efficiency to DNA substrate containing 5 sugar phosphate group in the nick/gap, whereas DNA polymerase beta binds to DNA duplex with a mononucleotide gap flanked by the 5 -p group. When APE1 and DNA polymerase beta are both present, a ternary complex APE1-DNA polymerase beta-DNA is formed with the highest efficiency with DNA product of APE1 endonuclease activity and with DNA containing 5 -flap or mononucleotide-gapped DNA with 5 -p group. It was found that APE1 stimulates DNA synthesis catalyzed by DNA polymerase beta, and a human X-ray repair cross-complementing group 1 protein (XRCC1) stimulates APE1 3 -5 exonuclease activity on 3 -recessed DNA duplex. PMID- 18393761 TI - Ribonuclease, deoxyribonuclease, and antiviral activity of Escherichia coli expressed Bougainvillea xbuttiana antiviral protein 1. AB - A full-length cDNA encoding ribosome-inactivating/antiviral protein from the leaves of Bougainvillea xbuttiana was recently isolated. The coding region of cDNA was cloned and expressed in Escherichia coli, and the protein product was designated as BBAP1 (Bougainvillea xbuttiana antiviral protein 1). BBAP1 showed ribonuclease activity against Torula yeast RNA. It also exhibited depurination activity against supercoiled pBlueScript SK+ plasmid DNA in a concentration dependent manner, and was found to convert nicked circular DNA into linear form only at higher concentration. On bioassay, BBAP1 exhibited antiviral activity against sunnhemp rosette virus infecting Cyamopsis tetragonoloba leaves in which 95% inhibition of local lesion formation was observed. PMID- 18393762 TI - Membrane localization of the MAK-V protein kinase. AB - Activities of many proteins including protein kinases are often regulated by their dynamic association with specific intracellular compartments. MAK-V is an AMPK-like protein kinase with poorly characterized functions and mechanisms of action. Similarly to many other protein kinases, association of MAK-V with specific intracellular compartments could be essential for its proper functions. In this work, we studied subcellular distribution of exogenously produced and endogenous MAK-V proteins in mammalian cells using biochemical cell fractioning aiming to supplement data on MAK-V intracellular localization studied by immunocytochemical methods. We found that a significant portion of MAK-V protein in mammalian cells is associated with membranes. Moreover, MAK-V expressed in yeast was also targeted to membrane, thus suggesting an evolutionarily conservative mechanism of MAK-V membrane association. Based on the ability of various MAK-V deletion mutants to localize to membrane and comparison of MAK-V amino acid sequences from different species, we suggest a possible mechanism governing MAK-V association with intracellular membranes. PMID- 18393763 TI - Lipid rafts association of synaptotagmin I on synaptic vesicles. AB - We confirmed the raft association of synaptotagmin I (syt I) in synaptic vesicles by sucrose density gradient centrifugation, cholesterol depletion, and temperature dependence, and Ca2+ was found to positively regulate this association. Furthermore, using syt I mutants we found that the transmembrane domain (TMD) of syt I plays an important role in localizing syt I into the lipid rafts of synaptic vesicles, and the raft association of the TMD can be regulated by its phosphorylation status. PMID- 18393764 TI - Isolation and physicochemical properties of tankyrase of human embryonic kidney cells of line 293. AB - We have isolated and purified endogenous cytosolic tankyrase from human embryonic kidney cells of line 293. Our data confirm a model of De Rycker and Price who consider that tankyrase is a master scaffolding protein capable of regulating assembly of large protein complexes. We have also studied kinetic characteristics of tankyrase in the complex, pH dependence of the enzyme activity, and its physicochemical properties. PMID- 18393765 TI - Lipid synthesis in macrophages during inflammation in vivo: effect of agonists of peroxisome proliferator activated receptors alpha and gamma and of retinoid X receptors. AB - The effects of peroxisome proliferator activated receptors alpha and gamma (PPAR alpha and PPAR-gamma) and retinoid X receptor (RXR) agonists upon synthesis and accumulation of lipids in murine C57Bl macrophages during inflammation induced by injection of zymosan and Escherichia coli lipopolysaccharide (LPS) have been studied. It is significant that intraperitoneal injection of zymosan (50 mg/kg) or LPS (0.1 mg/kg) in mice led to a dramatic increase of [14C]oleate incorporation into cholesteryl esters and triglycerides and [14C]acetate incorporation into cholesterol and fatty acids in peritoneal macrophages. Lipid synthesis reached its maximum rate 18-24 h after injection and was decreased 5-7 days later to control level after LPS injection or was still heightened after zymosan injection. In macrophages obtained in acute phase of inflammation (24 h), degradation of 125I-labeled native low density lipoprotein (NLDL) was 4-fold increased and degradation of 125I-labeled acetylated LDL (AcLDL) was 2-3-fold decreased. Addition of NLDL (50 microg/ml) or AcLDL (25 microg/ml) into the incubation medium of activated macrophages induced 9-14- and 1.25-fold increase of cholesteryl ester synthesis, respectively, compared with control. Addition of NLDL and AcLDL into the incubation medium completely inhibited cholesterol synthesis in control macrophages but had only slightly effect on cholesterol synthesis in activated macrophages. Injection of RXR, PPAR-alpha, or PPAR-gamma agonists--9-cis-retinoic acid (5 mg/kg), bezafibrate (10 mg/kg), or rosiglitazone (10 mg/kg), respectively--30 min before zymosan or LPS injection led to significant decrease of lipid synthesis. Ten hour preincubation of activated in vivo macrophages with the abovementioned agonists (5 microM) decreased cholesteryl ester synthesis induced by NLDL and AcLDL addition into the cell cultivation medium. The data suggest that RXR, PPAR-alpha, or PPAR-gamma agonists inhibited lipid synthesis and induction of cholesteryl ester synthesis in inflammatory macrophages caused by capture of native or modified LDL. PMID- 18393766 TI - Role of glucocorticoids and resident liver macrophages in induction of tyrosine aminotransferase. AB - Administration of cortisol to an animal induces tyrosine aminotransferase (TAT) in the liver. A similar effect was observed after stimulation of resident liver macrophages (Kupffer cells) by dextran sulfate. Actinomycin D completely blocks enzyme induction both by cortisol and dextran sulfate, whereas their combined effect gives an additive result. In primary culture of hepatocytes, dextran sulfate inhibits TAT activity, but conditioned macrophage medium reliably increases enzyme activity in hepatocytes. However, incubation of isolated macrophages in the presence of dextran sulfate and such medium transfer into hepatocyte culture results in even more pronounced increase in TAT activity. In a combined culture of hepatocytes and non-parenchymal liver cells, reproducing intercellular interactions in vitro, cortisol and non-parenchymal cells exhibit an additive effect on TAT activity. These results show that liver macrophages release a factor of unknown nature launching the mechanism of TAT induction independently of cortisol, a classic TAT inducer. PMID- 18393767 TI - Extracellular yeast-lytic enzyme of the bacterium Lysobacter sp. XL 1. AB - An enzyme exhibiting yeast-lytic activity has been isolated from the culture liquid of the bacterium Lysobacter sp. XL 1. The optimal conditions for the hydrolysis of Saccharomyces cerevisiae cells by the enzyme have been established: 0.15 M sodium acetate buffer, pH 6.0, 50 degrees C. The yeast-lytic activity of the enzyme is inhibited by EDTA, p-chloromercuribenzoate, and phenylmethylsulfonyl fluoride. According to the data of SDS-PAGE, the molecular weight of the protein is 36 kD. The enzyme hydrolyzes casein, hemoglobin, and synthetic peptide Abz-Ala-Ala-Phe-pNA, i.e. it exhibits proteolytic activity. The properties of the enzyme and its molecular weight correspond to those of a previously isolated extracellular metalloproteinase. The N-terminal amino acid sequence of the protein exhibits 67% homology with the N-terminal sequence of achromolysine of Achromobacter lyticus (EC 3.4.24.-). PMID- 18393768 TI - Proteins and peptides of the salivary gland secretion of medicinal leeches Hirudo verbana, H. medicinalis, and H. orientalis. AB - The protein and peptide composition of medicinal leech salivary gland secretion (SGS) was analyzed in preparations obtained in July from three species--Hirudo verbana, H. medicinalis, and H. orientalis. Two-dimensional electrophoresis (molecular mass 10-150 kD and pI 3-10) revealed no distinctions in the distribution of over 100 silver-stained proteins. Differences were noted only in intensity of 10 protein spots at 30-90 kD and pI 4.7-7.5. Mass spectrometric profiling of SGS of the three leech species using the Zip-Tip/golden chip scheme and cation-exchanging chips CM-10 revealed over 50 components in SGS of each of the three leech species. It was noted that 30-40% of the individual masses of the SGS of each leech species fall within the masses present in SGS of at least one of the two other species. This rather small part of the total mass may be indicative of a high polymorphism of amino acid sequences or a high frequency of posttranslational modifications of the SGS proteins and peptides. Calculation of Jacquard's coefficient showed that H. medicinalis and H. orientalis are closest to each other in SGS composition, which is consistent with data in the literature on the phylogenetic relationship between these two species of medicinal leech. Comparison of detected molecular masses with those of six known biologically active compounds produced by medicinal leeches revealed their uneven distribution in SGS of each of the three medicinal leech species. This opens prospects for using certain species of medicinal leech for targeted therapy of various pathologies. PMID- 18393769 TI - Mitochondrial nucleoside diphosphate kinase: mode of interaction with the outer mitochondrial membrane and proportion of catalytic activity functionally coupled to oxidative phosphorylation. AB - In the present study, we found that ionic interactions are not essential for the binding of nucleoside diphosphate kinase of liver mitochondria outer compartment to outer mitochondrial membrane and that the proportion of the enzyme activity involved in functional coupling with oxidative phosphorylation (we demonstrated the existence of functional coupling earlier) is only 17%. Additional evidence was obtained that functionally coupled activity of nucleoside diphosphate kinase is associated with the outer surface of mitochondria. Dextran (10%) did not increase functional coupling. The physiological importance of these effects is discussed. PMID- 18393770 TI - Brain cathepsin B cleaves a caspase substrate. AB - We show that an enzyme exists in rat brain capable of cleaving the caspase-3 specific peptide substrate Ac-DEVD-AMC at low pH. The enzyme shows properties of a cysteine protease and is localized, predominantly, in lysosomes. We have purified this enzyme from rat brain and identified it by MALDI-TOF MS. The enzyme possessing "acidic" DEVDase activity in rat brain appears to be cathepsin B. It remains obscure, whether cathepsin B participates in cleavage of caspase-3 substrates in vivo. We suggest that under certain conditions (e.g. in hypoxia) cathepsin B participates in cleavage of caspase-3 substrates in brain cells. PMID- 18393771 TI - Hyperactivation of succinate dehydrogenase in lymphocytes of newborn rats. AB - We measured the activity of mitochondrial succinate dehydrogenase (SDH) within cells, in media with near-physiological composition, in lymphocytes immobilized in a blood smear on glass. SDH activity was studied in newborn rats characterized by natural hyperadrenergic status and also in adult animals injected with epinephrine. In most newborns very high activities were recorded, which exceeded the activities in adults at rest 7-8-fold or 3-fold according to the conventional calculation, or more than 30- and 6-fold according to our more precise calculation. The findings support our concept about a selective interaction between adrenergic stimulation and oxidation of succinic acid. According to this concept, epinephrine and norepinephrine specifically activate oxidation of succinic acid, whereas blood micromolar concentrations of the latter stimulate the release of catecholamines (the receptor-mediated signaling effect). This interaction is half of a substrate-hormonal regulatory system responsible for connection of vegetative nervous system with oxidation in mitochondria of the innervated organs. The increase in succinate oxidation by catecholamines includes activation of the faster pathways of succinate generation than the complete Krebs cycle, in particular, the glyoxylate cycle that is shown in the newborn rats in the present study. PMID- 18393772 TI - siRNAs targeting mouse myostatin. AB - Eight different mouse myostatin small interfering RNA (siRNAs) were synthesized and tested. Five siRNAs showed a pronounced biological effect reducing myostatin mRNA content. For two of them, the myostatin mRNA level was reduced 3- and 4 fold, respectively. The obtained siRNAs can be used for study of biological effects of myostatin, both in vitro and in vivo. PMID- 18393773 TI - Evolution of DNA polymerase iota structure and function in eukaryotes. AB - Analysis of DNA polymerase iota (Pol iota) enzymic activity in different classes of eukaryotes has shown that error-prone activity of this enzyme can be found only in mammals, and that it is completely absent from organisms that are at lower stages of development. It was supposed that the emergence of the error prone Pol iota activity in mammals is caused by structural alteration of the active center. Possible functions of error-prone Pol iota in higher eukaryotes are discussed. PMID- 18393774 TI - Human heart LIM protein has transcription activation ability related to LIM domain 1. AB - Human heart LIM (hhLIM), a muscle-specific expressed transcriptional coactivator of cardiac hypertrophy related gene, stimulates transcriptional activation of the ANF gene promoter in H9C2 cells. We revealed that the N-terminal half of hhLIM mediated this activation, in which the LIM domain 1 and protein kinase C phosphorylation site are important, especially the LIM domain 1. Further, mutagenesis of the conserved Cys in the LIM domain 1 abolished its ability to activate cardiac hypertrophy. These findings suggest that hhLIM is a typical LIM family member with powerful transcription activation. PMID- 18393775 TI - Cytoskeleton inhibitors combined with TRAIL induce apoptosis in HeLa carcinoma cells overexpressing antiapoptotic protein Bcl-2. AB - TRAIL (Apo2L), a cytokine from the family of tumor necrosis factors (TNF), causes apoptosis in various types of tumor cells but is not toxic for normal cells. Recombinant TRAIL obtained using an original method stimulates the release of cytochrome c from mitochondria into the cytoplasm and apoptosis in HeLa carcinoma cells. Expression of oncoprotein Bcl-2 in these cells blocks both processes. The microtubule inhibitors taxol, nocodazole, and colcemid, as well as an inhibitor of actin microfilaments cytochalasin D, enhance the action of TRAIL and allow it to overcome protection caused by overexpression of Bcl-2. This effect is not associated with enhancement of early steps of TRAIL-dependent apoptosis leading to activation of caspase-8 and Bid protein. The inactivation of Bcl-2 also does not define the effect of cytoskeleton inhibitors. It is supposed that destruction of cytoskeleton alters the mechanism of the TRAIL- (or TNF)-dependent cytochrome c release from mitochondria by making it resistant to Bcl-2. The combined use of cytoskeleton inhibitors, which are antitumor drugs, with the recombinant TRAIL preparations may be efficient in therapy of tumors resistant to traditional chemotherapy. PMID- 18393777 TI - Recent clinical trials of cladribine in hematological malignancies and autoimmune disorders. AB - The purine nucleoside analog - cladribine (2-chlorodeoxyadenosine, 2-CdA) is a cytotoxic agent with high activity in lymphoid and myeloid malignancies. It is also an effective drug in some autoimmune disorders. 2-CdA is usually administered intravenously in continuous or 2-hour infusion. Recently however, new formulation of this agent has been developed for subcutaneous and oral administration. 2-CdA is widely established as first line standard treatment for hairy cell leukemia. Moreover several clinical trials have demonstrated that this agent, used alone or in combination with other cytotoxic drugs, showed good efficacy and acceptable toxicity profile in the treatment of chronic lymphocytic leukemia, Waldenstrom macroglobulinemia, low-grade non-Hodgkin's lymphoma and acute myeloid leukemia. Moreover, some studies indicate that 2-CdA has some activity in progressive multiple sclerosis and other autoimmune disorders including autoimmune hemolytic anemia, rheumatoid arthritis, systemic lupus erythematosus, psoriasis and in patients with refractory factor VIII inhibitors. This review article will summarize the results of recent clinical trials with 2 CdA in hematological malignancies, multiple sclerosis and other autoimmune diseases. PMID- 18393776 TI - Epidermal growth factor receptor (EGFR) targeted therapies in non-small cell lung cancer (NSCLC). AB - The Epidermal Growth Factor Receptor (EGFR) family, including EGFR, HER2, HER3, and HER4, is implicated in the development and progression of cancer, and is expressed in many human epithelial malignancies, including Non-Small Cell Lung Cancer (NSCLC). Several molecules were synthesized to inhibit the extracellular domain of EGFR, such as cetuximab (Erbitux), the extracellular domain of HER2, such as trastuzumab (Herceptin) or the EGFR tyrosine kinase domain, such as gefitinib (Iressa) and erlotinib (Tarceva). Gefitinib and erlotinib are orally active, selective EGFR tyrosine-kinase inhibitors (EGFR-TKI) that produce objective response rates in about 10% of advanced NSCLC. More recently, erlotinib produced a significant improvement in survival when compared to placebo in pretreated NSCLCs. Among clinical characteristics, although female gender, and adenocarcinoma histology, showed to be significantly associated to TKI sensitivity, never smoking history is probably the most relevant factor. Presence of specific EGFR gene mutations or EGFR gene amplification confer a particularly sensitive phenotype, and patients with activation of the anti-apoptotic protein Akt are more sensitive, when Akt activation is sustained by a EGFR dependent mechanism. Cetuximab is a human-murine chimeric anti-EGFR IgG monoclonal antibody that has demonstrated both in vitro and in vivo antitumor activity in tumor cell lines expressing EGFR. It has shown impressive activity when combined with radiation by increasing the antitumor effect of radiation therapy. Cetuximab has a synergistic effect with cisplatin and may play a role in reversing resistance to chemotherapy. Cetuximab demonstrated to be active in pretreated NSCLCs, and its activity as first-line therapy in combination with chemotherapy is currently under evaluation. Efforts should be made for the identification of biological mechanism underlying cetuximab sensitivity and emerging data suggest that the drugs is more active in patients with EGFR gene amplification. In NSCLC, trastuzumab produced disappointing results when combined with chemotherapy, but probably patients were not properly selected. Recent findings in gefitinib treated patients support HER2 analysis by fluorescence in situ hybridization as a complementary test for selection of patient candidate for EGFR targeted therapies. Combination of EGFR targeting agents with other biological drugs is under investigation. PMID- 18393778 TI - Medical treatment of gastrointestinal stromal tumors: state of the art and future perspectives. AB - Gastrointestinal Stromal Tumor (GIST) is the most common mesenchymal neoplasm of the gastrointestinal tract, and it is characterized by the occurrence, in > 90 % of cases, of a gain of function mutation in the c-kit proto-oncogene. STI-571 (imatinib mesylate), a selective KIT tyrosine kinase inhibitor, has changed the natural history of this disease, since it has shown high effectiveness in metastatic GIST, and it is currently under investigation also in the adjuvant and neoadjuvant setting. Mechanisms of resistance to imatinib mesylate include both de novo, and, more frequently, acquired resistance, which may occur after several months of drug administration and possibly depends, in most cases, upon an acquired second mutation. In order to overcome imatinib mesylate resistance, the addition of other drugs may be considered in patients who have less than an optimal response to imatinib mesylate monotherapy. Investigational agents that are being studied in this setting include the mammalian target of rapamycin (mTOR) inhibitor RAD 001 and the protein kinase C inhibitor PKC412. In addition, other KIT tyrosine kinase inhibitors with anti-VEGF receptor inhibitory activity, such as SU11248, PTK787/ZK787 and AMG 706, are currently being explored as second line monotherapy for imatinib mesylate-resistant GIST. Finally, another new drug, ecteinascidin (ET-743), that blocks cell cycle progression in G2/M phase through a p53-independent apoptotic mechanism, has shown important preclinical and clinical activity against a number of human solid tumors, including GIST. PMID- 18393779 TI - Combined modality treatment of glioblastoma multiforme: the role of temozolomide. AB - Despite of improvements in the biological and molecular characterization of glioblastoma multiforme and studies of factors associated with tumor growth and progression, this type of malignant astroglial brain tumor is still difficult to treat. The present article reviews established and emerging prognostic and predictive factors and their potential influence on future therapeutic efforts. Recent developments in standard treatment options (surgery, radiotherapy and chemotherapy) are summarized. The integration of the oral cytotoxic agent temozolomide into current treatment protocols of postoperative combination therapy with radiation and drugs is discussed, especially in the context of the recently published randomized trial of the EORTC/NCIC, which showed that radiotherapy plus concomitant and adjuvant temozolomide significantly improved progression-free and overall survival over radiotherapy alone. The study also provided hypotheses about the subgroups, which are most likely to benefit from this reasonably well tolerated regimen. In a subset of patients, investigation of MGMT promoter methylation in tumor tissue was performed. Survival was shorter in patients with unmethylated promoter in both study groups. Patients with methylated promoter treated with radiotherapy had a median survival of 15 months, those treated with radiation plus temozolomide of 22 months (p=0.007). In the unmethylated group, the difference in median survival was only 1 month (p=0.06). Especially for these patients, alternative treatments need to be developed. The optimum schedule of temozolomide administration and the influence of combinations with additional antineoplastic agents remains to be studied. Early results of clinical trials addressing these issues are presented. PMID- 18393780 TI - Topical therapy for actinic keratoses: current and evolving therapies. AB - Actinic keratoses (AKs) are evolving malignant cutaneous neoplasms. They are also known as solar keratosis, squamous cell carcinoma in situ-solar keratotic type, or keratinocytic intraepidermal neoplasia. Actinic keratoses can be treated by two general methods: by physical/destructive methods and with topical therapies. This article will review current and evolving topical therapeutic options for AKs. Several topical treatment options have been shown to offer some significant benefit in the alleviation of these lesions. The therapies include 5 fluorouracil, imiquimod, diclofenac, colchicine and retinoids. PMID- 18393781 TI - The development of future research strategies from reviewing antiemetic trials for chemotherapy induced emesis. AB - In reviewing the latest trials of antiemetic usage to prevent cytotoxic chemotherapy induced emesis, gaps in the literature suggest directions for future research and identify methodological approaches to be used in future investigations. The usage of molecular techniques and the identification of new receptors may allow new antiemetics to be developed and identification of the genes coding for antiemetic receptors may be used to select the appropriate antiemetics for individuals. Given the success achieved in controlling post chemotherapy vomiting, future studies should focus upon the control of nausea, and measure the impact of antiemetic control on quality of life as well as evaluating the pharmacoeconomics of these agents. Accounting for the interaction of antiemetics with cytotoxics becomes more important in trial design with the increasing complexity of antiemetic regimens. More information is needed on the emetic potential of the various combination chemotherapy regimens, multiple day chemotherapy and chemotherapy over multiple cycles. The emetic potential of prolonged administration of oral chemotherapy and newer biologicals and targeted therapies needs to be recorded. Further studies are required in specialized areas such as with high dose chemotherapy, for radiation induced emesis and in pediatrics. PMID- 18393782 TI - Clinical benefit of idiotype vaccines: too many trials for a clever demonstration? AB - The most accepted and least biased manner to demonstrate clinical benefit for any new treatment is to show that it conveys a survival advantage in a well-designed phase III, randomized clinical trial. However, in selected cases, an exception can be made to this sound rule. This review aims at elucidating one such example. In particular, I intend to show that when an individualized form of immunotherapy like idiotypic vaccination, which by definition is completely inactive against any tumor cells, is applied to cancer patients with indolent follicular lymphoma, a carefully crafted phase II clinical trial may be able to demonstrate clinical benefit better and more rapidly than a phase III alternative. This consideration might be rather important over the next two to three years, since the results of as many as three ongoing phase III clinical trials on idiotype vaccines are expected to be unveiled within this time frame, following the release of conclusive data of our phase II clinical trial, which is imminent. PMID- 18393783 TI - Functional recovery after stroke: a review of current developments in stroke rehabilitation research. AB - This review article discusses current research developments in functional recovery after stroke. With the institutionalization of stroke services across health care facilities, a reduction in mortality rates, length of inpatient stay and improved independence in activities of daily living has been reported. Several systematic reviews show that traditional treatment approaches induce improvements that are confined to impairment level only and do not generalize to a functional improvement level. More recently developed treatment strategies that incorporate compensation strategies with a strong emphasis on functional training, may hold the key to optimal stroke rehabilitation. Intensity and task specific exercise therapy are important components of such an approach. Guidelines may assist the clinician in this responsibility. However, due to marked heterogeneity of the stroke population and poor methodological quality of many studies, results are uncertain. Several options are discussed to overcome the problem of stroke heterogeneity in research designs. Longitudinal repeated measurements designs are required to study the effects of non-linearity and time dependency of functional recovery in stroke. Furthermore, prognostic research based on sound clinimetric data generates relevant information that may guide the clinician in clinical decision making and in determining optimal treatment strategies. PMID- 18393784 TI - A review of methods for ensuring the comparability of comparison groups in randomized clinical trials. AB - While different design features of medical studies ostensibly serve different functions, many fall under the umbrella of methods aimed at ensuring the comparability of the comparison groups. Randomization rightly occupies the top spot in the hierarchy of design types, as it eliminates some biases (that is, systematic differences in comparison groups) that no other design can claim to eliminate. It is often assumed, and sometimes even asserted explicitly, that randomization by itself suffices to ensure that the comparison groups are sufficiently comparable that they would differ only randomly, but two points need to be made in this context. First, the assertion is not true. Second, even if it were true, it would still not be a cause for complacency, because even random baseline imbalances can wreck havoc on the valid interpretation of randomized clinical trials. Additional methods, beyond randomization, are therefore seen to be essential to the design of a good randomized clinical trial. Such methods include masking, allocation concealment, restrictions on the randomization, adjustment for prognostic variables, and the intent-to-treat approach to data analysis. Masking aims to ensure that those individuals in any one group formed by randomization are treated as similarly as possible subsequent to randomization as those in any other group formed by randomization. In contrast, allocation concealment and restricted randomization aim to create groups that start off as comparable. Adjustment for prognostic variables aims to change the comparison groups themselves to make them comparable. For example, one might find gender to be both predictive of outcome and unbalanced across treatment groups, and so one would compare the treatment groups not overall but rather first only among females and second only among males. The intent-to-treat approach aims to keep similar groups similar by not allowing for patient selection based on post randomization outcomes (including failure to comply with the protocol). The key to understanding masking, allocation concealment, and randomization is to recognize that none of them are binary phenomena, even though they are often incorrectly understood to be. So one must question how these methods are actually carried out, rather than contenting oneself with the vague statement that these methods were performed. This review will shed light on the distinction between the process and the outcome of each of these methods (masking, allocation concealment, and randomization), and will also consider issues related to adjustment for prognostic covariates. PMID- 18393785 TI - Porphyrin-carbohydrate conjugates: impact of carbohydrate moieties in photodynamic therapy (PDT). AB - In recent years, a number or review articles describing the utility of porphyrin based compounds in photodynamic therapy have been published. Currently, efforts are also underway in various laboratories to increase the tumor-specificity by developing target-specific photosensitizers. One of such attempts has been to synthesize a series of carbohydrate-porphyrin conjugates and investigate their ability to target carbohydrate- recognized proteins, which are known for their high expression in certain tumors. The present article is focused on the synthesis and biological significance of carbohydrate conjugated photosensitizers. PMID- 18393786 TI - Recent advances in two-photon photodynamic therapy. AB - Photodynamic therapy (PDT) is a treatment for tumors and accepted in several countries in the world. Introduction of two-photon absortion (2PA) into PDT allows spatially selective treatment of cancers. Possibilities and limitations of the use of two-photon excitation in PDT are discussed, and many literatures in this area are reviewed. The conclusions are that 2PA-PDT has an advantage for higher selectivity than one-photon absorption PDT, and femtosecond pulsed laser is more suitable for 2PA-PDT than pico- and nanosecond pulses. However, most of photosensitizers used in the past studies had low 2PA cross section values less than 50 GM, and resulted in a low PDT efficiency under two-photon irradiation conditions. To realize 2PA-PDT, much larger 2PA cross sections must be required. PMID- 18393787 TI - Photodynamic therapy: the development of new photosensitisers. AB - The first 20 years of anticancer photodynamic therapy (PDT) were based on the utility of the oligomeric mixture haematoporphyrin derivative (HpD) in various forms. More recently new derivatives have become available, both porphyrin derived and employing new chromophores, for example from the phthalocyanine and phenothiazinium families. In addition, a major research effort has been rewarded with the clinical acceptance of the porphyrin precursor 5-aminolaevulinic acid (ALA). New photosensitisers intended for clinical use must exhibit advantageous drug performance profiles compared to the first-generation porphyrin derivatives. This can be seen, in vitro, in improved photophysical properties such as the extension of the useful light absorption spectrum into the near infrared - offering greater tissue penetration - as well as in the synthesis of pure compounds rather than mixtures. In this review, recent developments in photosensitiser families are discussed with respect to in vitro performance indicators and to potential application in oncology. PMID- 18393788 TI - Diazo-containing molecular constructs as potential anticancer agents: from diazo[b]fluorene natural products to photoactivatable diazo-oxochlorins. AB - DNA is an established biological target for many organic natural products that react by alkylation or H-atom abstraction via key functional groups such as cyclopropane, aziridine, enediyne, and terminal diazo functionalities. Remarkably, although more than 20 natural product derivatives compose the latter class, the precise mechanism of action and specific biological target remain to be elucidated. Despite these biochemical uncertainties, more than 100 years of diazo/diazoketone chemistry exists. Much of this work involves photochemical N(2) extrusion to generate an initial carbene intermediate capable of insertion (singlet), H-atom abstraction (triplet), or ketene formation and subsequent nucleophilic addition (Wolff rearrangement). The trigger advantage of photochemical reactivity, coupled with the entropic gain of deazetation, and the high reactivity of the resulting intermediate, have led researchers to consider diazo compounds as potential phototherapeutric agents for medical applications. Such a strategy could serve as an alternative to (1)O(2) generation in photodynamic therapy (PDT), particularly in solid tumors or other hypoxic environments. Since diazoparaquinone natural products, and diazo compounds in general, are susceptible to redox activated N(2) loss, transition metal complexes containing redox-active excited states that absorb in the tissue transparent therapeutic window have potential as new therapeutic agents. Moreover, highly pi conjugated molecules such as porphyrins and chlorins, which serve as the primary pigment for current PDT due to intense absorption bands throughout the region of 600-850 nm, have only recently been able to support a conjugated diazoketone functionality at the macrocycle periphery. These synthetic advances have now made diazo activation through visible region photolysis possible, and have led to characterization of a range of remarkable molecular photoproducts including azeteoporphyrinoids and O-H/N-H insertion products. In addition to protein or DNA alkylation, the latter reactivity leads to potential for these constructs to serve as in situ biological labels or as recognition elements to probe biochemical mechanisms. PMID- 18393789 TI - The warburg effect: why and how do cancer cells activate glycolysis in the presence of oxygen? AB - Cells can obtain energy through the oxygen-dependent pathway of oxidative phosphorylation (OXPHOS) and through the oxygen-independent pathway of glycolysis. Since OXPHOS is more efficient in generating ATP than glycolysis, it is recognized that the presence of oxygen results in the activation of OXPHOS and the inhibition of glycolysis (Pasteur effect). However, it has been known for many years that cancer cells and non-malignant proliferating cells can activate glycolysis in the presence of adequate oxygen levels (aerobic glycolysis or Warburg effect). Accumulating evidence suggests that the persistent activation of aerobic glycolysis in tumor cells plays a crucial role in cancer development; the inhibition of the increased glycolytic capacity of malignant cells may therefore represent a key anticancer strategy. Although some important knowledge has been gained in the last few years on this growing field of research, the basis of the Warburg effect still remains poorly understood. This communication analyzes why cancer cells switch from OXPHOS to glycolysis in the presence of adequate oxygen levels, and how these cells manage to avoid the inhibition of glycolysis induced by oxygen. Several strategies and drugs that may interfere with the glycolytic metabolism of cancer cells are also shown. This information may help develop anticancer approaches that may have clinical relevance. PMID- 18393790 TI - Cancer chemoprevention by garlic and its organosulfur compounds-panacea or promise? AB - Of late medicinal plants and functional foods rich in bioactive phytochemicals have received growing attention as potential agents for cancer chemoprevention. Accumulating evidence from epidemiological studies as well as laboratory data supports the anticancer properties of garlic widely used as a medicinal herb and spice. Garlic and its organosulfur compounds (OSCs) appear to exert their anticarcinogenic effects through multiple mechanisms that include modulation of carcinogen metabolism, inhibition of DNA adduct formation, upregulation of antioxidant defences and DNA repair systems, and suppression of cell proliferation by blocking cell cycle progression and/or inducing apoptosis. Since multiple signaling pathways are dysfunctional in cancer and new oncogenic mutations accumulate with carcinogenic progression, dietary agents such as garlic with its rich array of bioactive OSCs that modulate cancer cascades offer promise as potential chemopreventive and chemotherapeutic agents. PMID- 18393791 TI - Caspase activity modulators as anticancer agents. AB - Proteolytic caspase enzymes play a central role in cell apoptosis, or programmed cell death, often as integrating elements of different stimuli leading to the cell death. Since blockade of apoptotic pathways are fundamental for cell survival and proliferation, particularly in cancer cells, the activation of caspases is an attractive target for anticancer therapy. This review describes some of the druggable therapeutic targets thus far identified within the core apoptotic machinery, the corresponding drugs that have been developed, their effects on caspase-dependent apoptotic pathways and their potential impact on the therapy of cancer. With several successful anticancer drugs on the market and numerous compounds in preclinical and clinical developments, modulators of caspase-dependent apoptotic pathways belong to the most important category of anticancer agents. PMID- 18393792 TI - Inhibition of protein kinase c-Src as a therapeutic approach for cancer and bone metastases. AB - c-Src is a proto-oncogene involved in the genesis of and invasion by many cancers. This non-receptor tyrosine kinase also plays a crucial role in bone homeostasis, since inhibition or deletion of c-Src impairs the function of osteoclasts, the bone resorbing cells. It is thus conceivable that c-Src could be a suitable target for the pharmacological treatment of cancers, skeletal metastases and diseases of bone loss, such as osteoporosis. The pyrrolo pyrimidines CGP77675 and CGP76030 proved to be effective in preventing bone loss in animal models, while the effect of AZD0530, a dually active inhibitor of c-Src and Bcr-ABL, on bone resorption, has been tested in a Phase I clinical trials with promising results. As far as the metastatic bone disease is concerned, c-Src inhibitors could potentially have inhibitory effects both on osteoclasts and on tumour cells, and could disrupt the vicious circle established between these cell types in the bone microenvironment. In accord with this idea, CGP76030 is able to reduce the incidence of osteolytic lesions and of visceral metastases, and to suppress morbidity and lethality in a bone metastasis mouse model without obvious adverse effects. The purine-based c-Src inhibitor AP23451 and the dual c-Src/Abl inhibitors AP22408 and AP23236 proved efficacious in reducing bone metastases in preclinical studies. These results open a new avenue for the development of innovative therapies for the treatment of bone metastatic disease. PMID- 18393793 TI - Recent advances in cancer chemotherapy- part II. PMID- 18393794 TI - Production and fate of amyloid peptides: recent advances and perspectives. PMID- 18393795 TI - Processing of amyloid precursor protein and amyloid peptide neurotoxicity. AB - Alzheimer's disease is characterized by the presence of two types of lesions in brain: neurofibrillary tangles and senile plaques. Intraneuronal neurofibrillary tangles are made of paired helical filaments containing hyperphosphorylated microtubule associated protein tau. Extracellular senile plaques contain a core of beta-amyloid peptide (Abeta), which is produced by cleavage of the Amyloid Precursor Protein (APP). Among the two catabolic pathways of APP, the amyloidogenic pathway producing Abeta peptides was intensively studied in different cellular models expressing human APP. Differences in APP processing and in toxicity resulting from Abeta accumulation can be observed from one cell type to another. In particular, primary cultures of neurons process APP differently compared with other cultured cells including neuronal cell lines. Neurons accumulate intraneuronal Abeta, which is neurotoxic, and in these cells, APP can be phosphorylated at specific residues. Recent studies suggest that APP phosphorylation can play an important role in its amyloidogenic processing. In addition, protein kinases that phosphorylate APP are also able to phosphorylate the neuronal protein tau. Biochemical analysis of these two proteins in primary cultures of neurons show that phosphorylation of both APP and tau can be a factor linking the two characteristic lesions of Alzheimer's disease. PMID- 18393796 TI - BACE1 structure and function in health and Alzheimer's disease. AB - Amyloid plaques, hallmark neuropathological lesions in Alzheimer's disease (AD) brain, are composed of the beta-amyloid peptide (Abeta). Much evidence suggests that Abeta is central to the pathophysiology of AD and is likely to play an early role in this intractable neurodegenerative disorder. Given the strong correlation between Abeta and AD, therapeutic strategies to lower cerebral Abeta levels should prove beneficial for AD treatment. Abeta is derived from amyloid precursor protein (APP) via cleavage by two proteases, beta- and gamma-secretase. The beta secretase has been identified as a novel aspartic protease named BACE1 (beta-site APP Cleaving Enzyme 1) that initiates Abeta formation. Importantly, BACE1 appears to be dysregulated in AD. As the rate-limiting enzyme in Abeta generation, BACE1, in principle, is an excellent therapeutic target for strategies to reduce the production of Abeta in AD. While BACE1 knockout (BACE1-/-) mice have been instrumental in validating BACE1 as the authentic beta-secretase in vivo, data indicates that complete abolishment of BACE1 may be associated with specific behavioral and physiological alterations. Recently a number of non-APP BACE1 substrates have been identified. It is plausible that failure to process certain BACE1 substrates may underlie some of the reported abnormalities in the BACE1-/- mice. Here we review the basic biology of BACE1, focusing on the regulation, structure and function of this enzyme. We pay special attention to the putative function of BACE1 during normal conditions and discuss in detail the relationship that exists between key risk factors for AD and the pathogenic alterations in BACE1 that are observed in the diseased state. PMID- 18393797 TI - Memapsin 2 (beta-secretase) inhibitors: drug development. AB - Memapsin 2 (beta-secretase, BACE 1) processing of beta-amyloid precursor protein is the first step in the pathway leading to the production of amyloid-beta, thus, it is a major target for the development of inhibitor drug for the treatment of Alzheimers's Disease. Although there are distinctive advantages of this protease as a drug target, the development of drug-like memapsin 2 inhibitors has been somewhat slow since the cloning of the protease seven years ago. Here we review the progress of memapsin 2 inhibitor development using crystal structure-based design cycles. Recent progress has evolved the inhibitors into sizes sufficiently small to penetrate cell membranes and the blood-brain barrier yet retain potency for the inhibition of Abeta production in cultured cells and experimental animals. Such progress lends optimism that clinically useful memapsin 2 inhibitors will eventually be developed. PMID- 18393799 TI - The catalytic core of gamma-secretase: presenilin revisited. AB - Mutations in the presenilin 1 (PS1) gene are the major cause of familial Alzheimer s disease (AD). They effect an increased production of the highly neurotoxic 42 amino acid variant of the amyloid-beta peptide (Abeta), which is believed to initiate the disease. Abeta is the product of two consecutive cleavages of the beta-amyloid precursor protein (APP) by two proteases, beta secretase and gamma-secretase. The latter enzyme has been identified as an intramembrane-cleaving multiprotein complex that apart from APP cleaves a large number of other type I transmembrane proteins. PS1 and its homologue PS2 are essential for gamma-secretase cleavage and more than a decade after their discovery it is now firmly established that they function as catalytic subunits of gamma-secretase. This review recapitulates the findings that led to this conclusion as well as the further progress made on the function of PS as gamma secretase since then. PMID- 18393798 TI - Assembly, maturation, and trafficking of the gamma-secretase complex in Alzheimer's disease. AB - In this review, we discuss the biology of gamma-secretase, an enigmatic enzyme complex that is responsible for the generation of the amyloid-beta peptide that constitutes the amyloid plaques of Alzheimer's disease. We begin with a brief review on the processing of the amyloid precursor protein and a brief discussion on the family of enzymes involved in regulated intramembrane proteolysis, of which gamma-secretase is a member. We then identify the four major components of the gamma-secretase complex - presenilin, nicastrin, Aph-1, and Pen-2 - with a focus on the identification of each and the role that each plays in the maturation and activity of the complex. We also discuss two new proteins that may play a role in modulating the assembly and activity of the gamma-secretase complex. Next, we summarize the known subcellular locations of each gamma secretase component and the sites of gamma-secretase activity, as defined by the production of Abeta. Finally, we close by synthesizing all of the included topics into an overarching model for the assembly and trafficking of the gamma-secretase complex, which serves as a launching point for further questions into the biology and function of gamma-secretase in Alzheimer's disease. PMID- 18393801 TI - Epsilon-secretase: reduction of amyloid precursor protein epsilon-site cleavage in Alzheimer's disease. AB - The accumulation and deposition of fibrillar Abeta is thought the primary cause of Alzheimer's disease (AD). Abeta is generated by sequential proteolytic processing involving beta- and gamma-secretase on Amyloid beta protein precursor (APP). Recently, gamma-secretase was shown to cleave near the cytoplasmic membrane boundary of APP, called epsilon-site cleavage, as well as in the middle of the membrane domain, called gamma-site cleavage. Recent findings indicate that gamma- and epsilon-site cleavage are regulated independently. In this review, the reduction of epsilon-site cleavage in AD and the importance of epsilon-site cleavage are discussed. PMID- 18393800 TI - Gamma-secretase inhibition and modulation for Alzheimer's disease. AB - Gamma-secretase is a multi-protein complex that proteolyzes the transmembrane region of the amyloid beta-peptide (Abeta) precursor (APP), producing the Abeta peptide implicated in the pathogenesis of Alzheimer's disease (AD). This protease has been a top target for AD, and various inhibitors have been identified, including transition-state analogue inhibitors that interact with the active site, helical peptides that interact with the initial substrate docking site, and other less peptide-like, more drug-like compounds. Although one gamma-secretase inhibitor has advanced into late-phase clinical trials, concerns about inhibiting this protease remain. The protease complex cleaves a number of other substrates, and in vivo toxicities observed with gamma-secretase inhibitors are apparently due to blocking one particularly important substrate, the Notch receptor. Thus, the potential of gamma-secretase as therapeutic target likely depends on the ability to selectively inhibit Abeta production without hindering Notch proteolysis (i.e., modulation rather than inhibition). The discovery of gamma secretase modulators has revived gamma-secretase as an attractive target and has so far resulted in one compound in late-phase clinical trials. The identification of other modulators in a variety of structural classes raise the hope that more promising agents will soon be in the pipeline. PMID- 18393802 TI - From presenilinase to gamma-secretase, cleave to capacitate. AB - Mutations in two genes, presenilin 1 (PS1) and its homologue presenilin 2 (PS2), account for a majority of early onset familial Alzheimer disease cases which are characterized by intracellular neurofibrillary tangles and extracellular amyloid fibrils composed of the amyloid beta protein (Abeta). Abeta is derived from sequential cleavages of Amyloid Precursor Protein (APP) by beta-secretase and gamma-secretase, the latter is composed of four components, PS1, nicastrin (NCT), presenilin enhancer 2 (PEN-2), and anterior pharynx defective (APH-1). These components not only maintain the stability of the gamma-secretase complex but also regulate the activity of presenilinase, the protease responsible for the cleavage of full length PS1 into N-terminal and C-terminal fragments (NTF/CTF). We have previously shown that endoproteolysis of PS1 into NTF/CTF by presenilinase requires two critical aspartate residues, suggesting that PS1 may undergo autoproteolysis; full length PS1 complexes with NCT, PEN-2, APH-1 and forms the presenilinase. While these two aspartate residues are necessary for the endoproteolysis of full length PS1, they are equally critical for the gamma secretase cleavage of multiple substrates, and it is hypothesized that the full length PS1/presenilinase is the zymogen of gamma-secretase. The inhibition profiles of presenilinase and gamma-secretase are illustrated by their biochemical similarity but are pharmacologically distinct. Since the uncleaved PS1 loop may obstruct the entry of gamma-secretase substrates to the docking site of the gamma-secretase complex, investigation of presenilinase inhibitors interfering with substrate-docking may facilitate a novel approach to identify APP specific gamma-secretase inhibitors. PMID- 18393803 TI - A closer look at alpha-secretase. AB - Accumulation of amyloid beta-peptides (Abeta) in the brain is believed to contribute to the development of Alzheimer disease (AD). Abeta, a 40-42 amino acid-comprising proteolytical fragment of the amyloid precursor protein (APP), is released from APP by sequential cleavages via beta- and gamma-secretases. However, the predominant route of APP processing consists of successive cleavages by alpha- and gamma-secretases. Alpha-secretase attacks APP inside the Abeta sequence, and therefore prevents formation of neurotoxic Abeta. After cleavage by alpha-secretase, the soluble N-terminal domain of APP, which possesses neurotrophic and neuroprotective properties, is released. In AD patients, a decrease in alpha-secretase processing of APP has been found and therefore, strategies to improve alpha-secretase activity are obvious. Several years after descriptive reports on alpha-secretase, the responsible enzymes have been identified to belong to the family of A Disintegrin And Metalloproteinase (ADAM). Three of these membrane-anchored zinc-dependent metalloproteinases, ADAM10 as well as ADAM17 and presumably also ADAM9 display alpha-secretase activity. Since the individual knock-out of these proteinases in neither case completely prevented alpha-secretase processing of APP, it seems likely that different ADAMs are compensating mutually, and under different conditions may contribute to alpha secretase cleavage of APP. In addition to ADAMs, perhaps other membrane associated metalloproteinases contribute to the shedding of APP. Stimulation of alpha-secretase activities can be achieved via several signaling cascades including phospholipase C, phosphatidylinositol 3-kinase and serine/threonine specific kinases such as protein kinases C, and mitogen activated protein kinases. Direct activation of protein kinase C and stimulation of distinct G protein-coupled receptors are known to increase alpha-secretase processing of APP. Agonists for M1 muscarinic acetylcholine receptors and serotonin 5-HT4 receptors are currently in clinical trials to test their efficiency in the treatment of AD. PMID- 18393804 TI - Part-time alpha-secretases: the functional biology of ADAM 9, 10 and 17. AB - Disintegrin metalloproteases of the ADAM family form a large (at present > 40 members in mammals) family of multidomain membrane proteins that in their ectodomain combine a cystein-rich, disintegrin and a zinc metalloprotease domain. Via their metalloprotease domain, ADAMs are often implicated in ectodomain shedding, either to release e.g. growth factors or to initiate further intracellular signalling via regulated intramembrane proteolysis. Mainly based upon overexpression studies in vehicle cells, three of them, ADAMs 9, 10 and 17, have been proposed to act as alpha-secretases for amyloid precursor protein (APP). It is striking thereby that this role has since then remained somewhat ill defined, as APP processing in ADAM9 deficient neurons is unaltered, and also ADAM10 deficient murine embryonic fibroblasts exhibit at best a highly variable reduction in alpha-secretase activity. However, during the past years, numerous other substrates have been linked to all three sheddases, the cleavage of which in some cases appears to be strikingly more important for the organism than APP processing. Most notably, the perinatally lethal phenotype of ADAM17 knockout mice is dominated by a loss of growth factor shedding, while the even earlier fatal effects of ADAM10 deficiency exhibit key features of disabled Notch signalling and possibly also cadherin processing defects. In this review, we will summarize the published data on the "non-APP" functions of all three ADAMs, the further evaluation of which may be crucial when attempting to treat Alzheimer s Disease by increasing their expression and/or activity. As the knockouts of ADAM10 and ADAM17 are only informative for their roles in (early) development, while a number of recently assigned new substrates play crucial roles in the normal and/or diseased adult organism as well, work on conditional knockout models will be crucial to fully characterize both the full functional portfolio of (candidate) alpha-secretases as well as their clinical relevance, which may go way beyond Alzheimer s Disease. PMID- 18393805 TI - Regulation of betaAPP and PrPc cleavage by alpha-secretase: mechanistic and therapeutic perspectives. AB - Alzheimer's disease (AD) is by far the most common form of dementia in the elderly and concerns one out of three individuals over 85. Like other neurodegenerative disorders such as Parkinson, Hungtington or prion diseases, AD is characterized by the formation of amyloid plaques in the central nervous system. In the brain of AD patients, the main component of these abnormal deposits is an aggregated form of the so-called amyloid beta-peptide (Abeta), which is produced from a large trans-membrane type-1 protein, the beta-amyloid precursor protein (betaAPP), by the sequential action of the beta- and gamma secretases. Beside these two amyloidogenic proteolytic attacks, betaAPP is targeted by a third enzyme termed alpha-secretase. Of utmost importance, this cleavage, which can be of constitutive or regulated origin, occurs right in the middle of the Abeta sequence, thus precluding its production. For this reason, and because the sAPPalpha secreted fragment derived from this cleavage displays beneficial effects, tremendous efforts have been made recently in order to both identify the proteases involved and the way they are regulated. More recently, it emerged that alpha-secretase was also responsible for the physiological processing of the cellular prion protein (PrP(c)) in the middle of its toxic 106 126 sequence. This review will focus on the recent advances in the alpha secretase pathways regulation and will discuss the putative therapeutic approaches that could be envisioned concerning the treatment of two apparently distinct diseases that share common denominators according to their metabolism. PMID- 18393806 TI - Amyloid-degrading enzymes as therapeutic targets in Alzheimer's disease. AB - The steady state concentration of the Alzheimer's amyloid-beta peptide in the brain represents a balance between its biosynthesis from the transmembrane amyloid precursor protein (APP), its oligomerisation into neurotoxic and stable species and its degradation by a variety of amyloid-degrading enzymes, principally metallopeptidases. These include, among others, neprilysin (NEP) and its homologue endothelin-converting enzyme (ECE), insulysin (IDE), angiotensin converting enzyme (ACE) and matrix metalloproteinase-9 (MMP-9). In addition, the serine proteinase, plasmin, may participate in extracellular metabolism of the amyloid peptide under regulation of the plasminogen-activator inhibitor. These various amyloid-degrading enzymes have distinct subcellular localizations, and differential responses to aging, oxidative stress and pharmacological agents and their upregulation may provide a novel and viable therapeutic strategy for prevention and treatment of Alzheimer's disease. Potential approaches to manipulate expression levels of the key amyloid-degrading enzymes are highlighted. PMID- 18393807 TI - Neprilysin and amyloid beta peptide degradation. AB - Neprilysin is a zinc metalloendopeptidase with relatively broad substrate specificity. The enzyme is localized to the plasma membrane of cells where it can function to degrade extracellular peptides. Structural studies show that neprilysin preferentially cleaves peptides on the amino side of hydrophobic amino acids. Neprilysin has been implicated in the catabolism of amyloid beta peptides in the brain and as such has received considerable attention, particularly as a therapeutic target for Alzheimer's disease. An inverse relationship between neprilysin levels and amyloid beta peptide levels and between neprilysin levels and amyloid plaque formation has been observed in human brain. Neprilysin levels decline with aging in the temporal and frontal cortex possibly contributing to higher amyloid beta peptide levels. A number of studies have shown that increasing neprilysin levels in the brain leads to a decrease in brain amyloid beta peptide levels. Most recently a potential relationship between amyloid beta peptide synthesis from the amyloid precursor protein and neprilysin activity has been proposed. PMID- 18393808 TI - Use of microwave in processing of drug delivery systems. AB - Microwave has received a widespread application in pharmaceuticals and food processing, microbial sterilization, biomedical therapy, scientific and biomedical analysis, as well as, drug synthesis. This paper reviews the basis of application of microwave to prepare pharmaceutical dosage forms such as agglomerates, gel beads, microspheres, nanomatrix, solid dispersion, tablets and film coat. The microwave could induce drying, polymeric crosslinkages as well as drug-polymer interaction, and modify the structure of drug crystallites via its effects of heating and/or electromagnetic field on the dosage forms. The use of microwave opens a new approach to control the physicochemical properties and drug delivery profiles of pharmaceutical dosage forms without the need for excessive heat, lengthy process or toxic reactants. Alternatively, the microwave can be utilized to process excipients prior to their use in the formulation of drug delivery systems. The intended release characteristics of drugs in dosage forms can be met through modifying the physicochemical properties of excipients using the microwave. PMID- 18393809 TI - Stimuli-responsive molecularly imprinted polymers for drug delivery: a review. AB - Molecular imprinting is an efficient technique for introducing regions with a highly specific molecular arrangement into a polymeric matrix. The first example of a molecularly imprinted polymer (MIP) was reported half a century ago; however, the use of molecular imprinting has become a well established practical tool only in the last decade. Recently, MIPs are widely used, for example, in chromatographic applications or enzyme antibody mimics. MIPs have also been used in biological applications such as drug delivery systems (DDS), and they have also been successfully applied as excipients in controlled delivery systems. Their huge potential could bring about intelligent drug release; this refers to the release, in a predictable way, of therapeutic agents in response to specific environmental stimuli (the presence of another molecule, pH changes, temperature, etc.). This review is focused on particular intelligent devices of this type that exhibit selective recognition (traps for toxic molecules) and release (of drugs in order to prolong the duration of pharmacological action) in response to specific stimuli. The "stimuli-responsive molecularly imprinted polymers" reviewed in this paper are expected to contribute significantly to the exploration and development of new generations of intelligent and self-regulated drug delivery systems. PMID- 18393810 TI - Gelrite microgels for sustained oral drug delivery-formulation and evaluation. AB - Gelrite is an ion-activated polymer prepared by partial acetylation of gellan gum, an exogeneous polysaccharide present in Pseudomonas elodea. Its unique cation-dependent gelling property at 33 degrees C has been exploited for the formulation of in situ gelling hydrogels for ophthalmic and oral drug delivery. Gelrite based oral formulations are known to sustain the drug levels at acidic pH due to the formation of 3D-lattice by cross-linking of uronic acid groups with cations. An extrusion method for formation of microspheres has been reported for Gelrite. The method cannot be successfully applied for encapsulation of hydrophilic moieties and has thus been modified in the present study. The microparticles prepared were optimized for various encapsulation variables such as particle size, shape, drug entrapment efficiency and in vitro drug release. A maximum of 41.87 +/- 0.2 % drug loading was obtained for the improved method. The drug release was found to be sustained in acidic medium (50% in SGF pH 1.2) for a period of 4 hours while burst release was observed in SIF pH 7.4. The work thus, presents a simplified microencapsulation method for hydrophilic drugs with higher entrapment efficiency for Gelrite, taking rifampicin as the model drug. PMID- 18393811 TI - Impregnation of an intraocular lens for ophthalmic drug delivery. AB - In this work the possibility of impregnating P(MMA-EHA-EGDMA) with flurbiprofen using a clean and environmentally friendly technology, namely supercritical fluid technology was evaluated. P(MMA-EHA-EGDMA) has been proposed as a promising matrix to be used for intraocular delivery of anti-inflammatory drugs used in eye surgery and flurbiprofen is a non-steroidal anti-inflammatory agent. Fundamental studies like, the solubility of the drug in carbon dioxide, as well as the sorption degree of this polymeric matrix in the presence of carbon dioxide have been previously carried out. The aim of this research was to evaluate the effects of these two variables in the impregnation process. Different experimental conditions were tested and the results obtained suggest that the best impregnating conditions for this system are low temperatures and pressures, which at the same time correspond to a lower solubility of the drug in the supercritical fluid and a low swelling of the polymeric matrix. Experiments performed also indicate that the batch impregnation process leads to higher yields of impregnation and according to the release profiles obtained the drug can be released from the matrix up to three months, which presents great advantages for post-surgical treatments. PMID- 18393812 TI - Nasal immunization studies by cationic, fusogenic and cationic-fusogenic liposomes encapsulated with tetanus toxoid. AB - Particulate antigens are more effective than soluble antigens in induction of systemic and mucosal immunity; possibly because they are more efficiently endocytosed by mucosal-associated lymphoid tissue (MALT) M cells. In this study, we determined the systemic and mucosal immune responses in rabbits following intranasal immunization with tetanus toxoid (TT) entrapped in cationic, fusogenic and cationic-fusogenic liposomes. Liposomes containing TT were prepared by dehydration-rehydration method. The volume mean diameter of cationic, fusogenic and cationic-fusogenic liposomes were 3.4 +/- 0.6, 4.3 +/- 2.3 and 3.4 +/- 1.5 microm, respectively. Encapsulation efficiency of TT in cationic, fusogenic and cationic-fusogenic liposomes was respectively determined as 49.1 +/- 8.4%, 48.5 +/- 2.1% and 50.8 +/- 4.9%. After 3 months, the leaking of encapsulated TT from liposomes ranged between 2.02 - 5.46%. Immunoreactivities of encapsulated TT in all kinds of liposomes were completely preserved, as studied by Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Enzyme-Linked Immunosorbent Assay (ELISA). The highest serum immunoglobulin G (IgG) and antitoxin titers were observed in groups immunized with solution formulation (P< 0.001). However, the highest mucosal secretory IgA (sIgA) titers were achieved by fusogenic liposomes (five times more titers compared with TT solution, and 15 times more titers compared with i.m. vaccine), followed by cationic-fusogenic liposomes. No hemolysis was occurred on incubation of liposomes and human erythrocytes. Also after nasal administration of plain liposomes to human volunteers, no local irritation was seen. This study suggests that intranasal administration of fusogenic and cationic-fusogenic liposomes encapsulated with vaccines could be an effective way for inducing mucosal immune responses. PMID- 18393813 TI - Advanced nebulizer designs employing vibrating mesh/aperture plate technologies for aerosol generation. AB - Recent technological advances and improved nebulizer designs have overcome many limitations of jet nebulizers. Newer devices employ a vibrating mesh or aperture plate (VM/AP) for the generation of therapeutic aerosols with consistent, increased efficiency, predominant aerosol fine particle fractions, low residuals, and the ability to nebulize even microliter volumes. These enhancements are achieved through several different design features and include improvements that promote patient compliance, such as compact design, portability, shorter treatment durations, and quiet operation. Current VM/AP devices in clinical use are the Omron MicroAir, the Nektar Aeroneb, and the Pari eFlow. However, some devices are only approved for use with specific medications. Development of "smart nebulizers" such as the Respironics I-neb couple VM technologies with coordinated delivery and optimized inhalation patterns to enhance inhaled drug delivery of specialized, expensive formulations. Ongoing development of advanced aerosol technologies should improve clinical outcomes and continue to expand therapeutic options as newer inhaled drugs become available. PMID- 18393814 TI - Novel hydrophilic drug polymer nano-conjugates of Cisplatin showing long blood retention profile: its release kinetics, cellular uptake and bio-distribution. AB - The present study evaluates the efficacy of drug polymer self folding nano conjugates of pectin-cisplatin to enhance blood circulating levels of cisplatin. The binding of nano-conjugate was confirmed by a peak-shift in UV-spectra. Physical characterization was done by DLS and TEM. Pharmacokinetics and bio distribution of the nano-conjugates were performed at various time points in normal, Balb-c mice. Zeta Potential showed the shielding effect on the negative potential of pectin that was approximately 7 times more than the pectin chains when conjugated with cisplatin. TEM confirmed the formation of a hydrophilic, easily re-dispersible nano-conjugate in the size range of 100 nm. Release kinetics in plasma showed that the pectin-cisplatin conjugate is a stable, slow and sustained system with no burst effect. Immuno-fluorescence analysis of J-774, a mouse macrophage cell line, was assessed after incubating the cells with pectin chains tagged with FITC as well as Pectin-Cisplatin-FITC conjugates. With the cellular uptake of these particles in J-774, 40% of the cells showed an uptake post 30 min of incubation. However, Pectin chains were clearly eliminated. The plasma proteins facilitate the release of cisplatin with 85-89% of the drug being released in 17 days, and only 57% of drug was released in approximately 30 days without plasma. The reduced negative charge on the conjugate helps in adhesion to the cell surface and subsequent uptake by cells as evidenced by cell uptake studies on J-774 cell line. Nano-conjugates showed long blood retention profile in mice and the cisplatin was found in circulation even after 24 hrs. Pharmacokinetic study clearly indicates that it can form a novel anticancer drug that possesses good efficacy and has a safer profile than cisplatin. PMID- 18393815 TI - Osmotically regulated flow of flurbiprofen through in situ formed asymmetric membrane capsule. AB - An in situ formed non-disintegrating controlled release asymmetric membrane capsular system, offering improved osmotic effect, was used to deliver poorly water soluble drug flurbiprofen (model drug) to demonstrate how controlled release characteristics could be manipulated by design of polymeric capsule with an asymmetric membrane. In situ formed asymmetric membrane capsule was made by dry method via precipitation of asymmetric membrane on the walls of hard gelatin capsule. Effect of different formulation variables were studied based on 2(3) factorial design, namely, level of osmogen, ethylcellulose and pore former apart from studying the effect of varying osmotic pressure on drug release. Scanning Electron Microscopy showed an outer dense non porous region and an inner lighter porous region for the prepared asymmetric membrane inside and a gelatin layer outside. Statistical test (Dunnett Multiple Comparison Test) was applied for in vitro drug release at P>0.05. The best formulation closely corresponded to the extra design checkpoint formulation by a similarity (f(2)) value of 96.88. The drug release was independent of pH but dependent on the osmotic pressure of the dissolution medium. The release kinetics followed Higuchi model and mechanism of release was Fickian diffusion. PMID- 18393816 TI - Preparation and evaluation of buccal bioadhesive tablets containing clotrimazole. AB - Buccal bioadhesive tablets of clotrimazole (CTZ) and clotrimazole: hydroxypropyl beta-cyclodextrin (CTZ-HPbetaCD) complex were prepared by using polymer xanthan gum in combination with carbopol 974P. The prepared buccal bioadhesive tablet formulations were evaluated for physicochemical characteristics (weight, hardness, friability, diameter, and drug content), swelling index, microenvironment pH, in-vitro drug release, bioadhesion strength, residence time and duration of antifungal activity (in-vitro). The dissolution of CTZ from the prepared tablets into phosphate buffer (pH 6.8) was controlled up to 8 h. All the prepared tablets gave reasonable in-vitro residence time (7.13 - 9.34 h). X-ray diffraction (XRD) studies of the CTZ-HPbetaCD complex, made by kneading and freeze-dried method, showed no CTZ crystal signals, demonstrating the inclusion of CTZ in the hydrophobic cavity of hydroxypropyl-beta-cyclodextrin (HPbetaCD) and formation of amorphous inclusion complex. Duration of the antifungal activity was measured by the inhibition zone of Candida albicans by agar diffusion assay. It is evident from the results obtained, the prepared buccal bioadhesive tablets of CTZ would markedly prolong the duration of the antifungal activity and may prove to be a viable alternative to the conventional local oral medication. PMID- 18393817 TI - The therapeutic index of locally acting inhaled drugs as a function of their fine particle mass and particle size distribution: a literature review. AB - The therapeutic index (TI) of locally acting inhaled drug products depends on a number of parameters and processes: the particle size distribution of the inhaled aerosol, the dose-efficacy response curves at the deposition sites, the amount of drug absorbed into the systemic circulation from the lung as well as the gastrointestinal (GI) tract, and the dose-effect curves for the different adverse drug reactions. In this review, we present qualitative scenarios, combining these effects and showing the possible influence of an envisaged change in the particle size distribution in the inhaled dose of a locally acting drug product on the TI. These scenarios are a valuable tool in the development of inhalation drug products. As a surrogate for the inhaled dose in vivo, we use the fine particle mass (FPM), measured by in vitro measurements. Using these scenarios, we reviewed the literature on bronchodilators and corticosteroids for reported associations between a change in the FPM and/or particle size distribution within the FPM, and the TI. We conclude that decreasing the particle size of an inhalation product may alter the TI both in a positive as well as a negative sense. So, smaller particle are not always better. PMID- 18393818 TI - Transdermal therapeutic system of enalapril maleate using piperidine as penetration enhancer. AB - The aim of this work was to formulate transdermal therapeutic system (TTS) of an antihypertensive drug, enalapril maleate (EM) using a new penetration enhancer, piperidine hydrochloride (PH), belonging to the class of Dihydropyridines. The TTS of EM was prepared by solvent evaporation technique using polymers Eudragit E100 and polyvinyl pyrrolidone K-30 in varying ratios, 5% w/w dibutylphthalate as plasticizer and 10% w/w PH as penetration enhancer. The TTS was evaluated for in vitro drug release using paddle over disc method and ex-vivo skin permeation using modified Keshary and Chein diffusion cell. The interaction studies were carried out by comparing the results of assay, UV and TLC analysis for pure drug and medicated and TTS formulation. Skin irritation potential of TTS was assessed by visual examination of treated rat skin. Stability studies were conducted according to ICH guidelines at a temperature of 40+/-0.5 degrees C and 75+/-5% RH. The optimized formulation was evaluated for preclinical bioavailability and antihypertensive efficacy using albino rat model. The optimized formulation provided 87.3% drug release in-vitro and a flux of 380 microg/cm(2)/hr over a period of 48 hours. No chemical interaction was found between the drug and excipients and there were no signs of skin irritation on application of patch. The optimized formulation was stable with a tentative shelf life of two years. Significant fall in BP (p<0.001) was observed in experimental hypertensive rats which was maintained for 2 days. There was 3 fold improvement in bioavailability with TTS vis-a-vis marketed tablet (AUC(0 to t) : 1253.9 ng.h/ml vs. 422.88 ng.h/ml). These preclinicial studies indicate the feasibility of matrix-type TTS of EM for 2 day management of hypertension. Further studies on human beings are warranted to establish clinical utility of the above TTS. PMID- 18393819 TI - Glycosyltransferases-part-I. PMID- 18393820 TI - Unique structural motif supports mannosylphospho dolichol synthase: an important angiogenesis regulator. AB - Mannosylphospho dolichol synthase (DPMS) catalyzes the transfer reaction GDP mannose + Dol-P <--> Dol-P-Man + GDP, a 'key step' in the assembly of lipid linked oligosaccharide (LLO) and a pre-requisite for asparagine-linked (N-linked) protein glycosylation. DPMS is present from a protozoan parasite to human, and its sequence carries a cAMP-dependent phosphorylation motif. We have evaluated the involvement of DPMS in angiogenesis, an essential physiological event during the growth of breast and other solid tumors. It has been observed that enhancers of intracellular cAMP accelerated the capillary endothelial cell proliferation by reducing the cell cycle duration. Reduced Con A to WGA fluorescence ratio indicated high level complex type N-glycans on the cell surface. This was supported by upregulated LLO biosynthesis in cells stimulated either with a beta agonist isoproterenol or other cAMP enhancer, such as 8Br-cAMP, forskolin, cholera toxin, or prostaglandin E1. The turnover (t((1/2))) of LLO was also increased. Increased LLO biosynthesis correlated extremely well with the DPMS activity in cells treated with 8Br-cAMP. High DPMS activity in isoproterenol treated cells was not due to an increased gene expression because actinomycin D failed to block the upregulation. cDNA cloning of capillary endothelial cell Dpm1 gene and the deduced amino acid sequence identified a PKA motif in capillary endothelial cell DPMS. Thus, it has been concluded that increased DPMS activity through protein phosphorylation is a driving force for angiogeneis. Its abolition, however, led to cell arrest in G1 and induction of apoptosis. PMID- 18393821 TI - Regulation of lactosylceramide synthase (glucosylceramide beta1-->4 galactosyltransferase); implication as a drug target. AB - Lactosylceramide is a ubiquitously present glycosphingolipid in mammalian tissues and has been implicated in cell proliferation, adhesion, migration and angiogenesis. This glycosphingolipid is synthesized by Golgi-localized enzyme LacCer synthase. According to recent nomenclature and gene mapping studies, two LacCer synthases beta1,4GalT-V and beta1,4GalT-VI have been identified and characterized. In addition, beta1,4GalT-V has been implicated in the synthesis of N-glycans of cell surface glycoproteins. During the past two decades data have accumulated suggesting that the cellular level of LacCer can be regulated by various growth factors, cytokines, lipids, lipoproteins and hemodynamic factors, such as fluid shear stress, by altering the activity of LacCer synthase. An interesting feature is that a nuclear regulating factor (SP1) plays a critical role in transcriptional regulation of this enzyme in cancer cells. Moreover, in human umbilical vein endothelial cells, NF-kappaB has been also shown to regulate this enzyme which, in turn, regulates the gene/protein expression of platelet endothelial cell adhesion molecule, intercellular cell adhesion molecule and angiogenesis. Since new blood supply via formation of capillaries is critical in tumor growth, metastasis, and atherogenesis, these findings expand the role of enzyme in these pathologies. Additional studies are warranted to understand the molecular and biochemical basis of how LacCer synthases are regulated. These studies will facilitate advances in discovery of drugs which mitigate diseases, such as atherosclerosis and cancer due to an aberrant regulation of these LacCer synthases. PMID- 18393822 TI - Golgi localization of glycosyltransferases involved in ganglioside biosynthesis. AB - Gangliosides make up a group of sialic acid-containing complex glycosphingolipids particularly abundant in the central nervous system. The finding indicating gangliosides are stored in certain hereditary diseases affecting the central nervous system opened the interest in studying their metabolism. The initial in vitro pioneering work on the glycosyltransferases involved in ganglioside biosynthesis was done by Roseman and his associates primarily in embryonic chick brains almost forty years ago. Since that time enzymes catalyzing the formation of main human gangliosides have been successfully purified and cloned. Their specificity has been determined and their subcellular localization and topology has been established. Transgenic mouse models deficient in distinct ganglioside directed glycosyltransferases are available and represent a vital step toward understanding the metabolism and function of this challenging lipid class. In the present review we briefly introduce the reader in the complex structure of gangliosides, then we summarize new developments concerning their function especially regarding neurodegenerative disorders, and in this article we would like to review on what is known about glycosyltransferases that catalyze the formation of these complex lipids in the Golgi apparatus, that was established by Basu and his associates almost three decades ago. PMID- 18393824 TI - Promoter structure and transcriptional regulation of human beta-galactoside alpha2, 3-sialyltransferase genes. AB - Six human beta-galactoside alpha2,3-sialyltransferase genes, which are hST3Gal I VI, have been cloned. Multiple genes encode enzymes with closely related catalytic specificities but different patterns of tissue expression. The multiple genes correspond to the control of various tissue specific regulators. Several studies have examined the transcriptional regulation of some human beta galactoside alpha2,3-sialyltransferases genes. Multiple mRNA forms differing only in the 5'-untranslated regions have been identified in hST3Gal II, hST3Gal III, hST3Gal IV, hST3Gal V, and hST3Gal VI. These transcripts are produced by a combination of alternative splicing and promoter utilization, suggesting the transcriptional regulation of this gene depends on the use of alternative promoters, further suggesting that tissue-specific transcriptional regulation of these genes depends on the use of multiple genes and multiple promoters. The multiple regulatory pathways of these ubiquitous sialyltransferases may be differentially modulated in various cell types. PMID- 18393823 TI - Structure and function of beta -1,4-galactosyltransferase. AB - Beta-1,4-galactosylransferase (beta4Gal-T1) participates in the synthesis of Galbeta1-4-GlcNAc-disaccharide unit of glycoconjugates. It is a trans-Golgi glycosyltransferase (Glyco-T) with a type II membrane protein topology, a short N terminal cytoplasmic domain, a membrane-spanning region, as well as a stem and a C-terminal catalytic domain facing the trans-Golgi-lumen. Its hydrophobic membrane-spanning region, like that of other Glyco-T, has a shorter length compared to plasma membrane proteins, an important feature for its retention in the trans-Golgi. The catalytic domain has two flexible loops, a long and a small one. The primary metal binding site is located at the N-terminal hinge region of the long flexible loop. Upon binding of metal ion and sugar-nucleotide, the flexible loops undergo a marked conformational change, from an open to a closed conformation. Conformational change simultaneously creates at the C-terminal region of the flexible loop an oligosaccharide acceptor binding site that did not exist before. The loop acts as a lid covering the bound donor substrate. After completion of the transfer of the glycosyl unit to the acceptor, the saccharide product is ejected; the loop reverts to its native conformation to release the remaining nucleotide moiety. The conformational change in beta4Gal-T1 also creates the binding site for a mammary gland-specific protein, alpha-lactalbumin (LA), which changes the acceptor specificity of the enzyme toward glucose to synthesize lactose during lactation. The specificity of the sugar donor is generally determined by a few residues in the sugar-nucleotide binding pocket of Glyco-T, conserved among the family members from different species. Mutation of these residues has allowed us to design new and novel glycosyltransferases, with broader or requisite donor and acceptor specificities, and to synthesize specific complex carbohydrates as well as specific inhibitors for these enzymes. PMID- 18393825 TI - Cloning and transcriptional regulation of genes responsible for synthesis of gangliosides. AB - Ganglioside synthases are glycosyltransferases involved in the biosynthesis of glycoconjugates. A number of ganglioside synthase genes have been cloned and characterized. They are classified into different families of glycosyltransferases based on similarities of their amino acid sequences. Tissue specific expression of these genes has been analyzed by hybridization using cDNA fragments. Enzymatic characterization with the expressed recombinant enzymes showed these enzymes differ in their donor and acceptor substrate specificities and other biochemical parameters. In vitro enzymatic analysis also showed that one linkage can be synthesized by multiple enzymes and one enzyme may be responsible for synthesis of multiple gangliosides. Following the cloning of the ganglioside synthase genes, the promoters of the key synthase genes in the ganglioside biosynthetic pathway have been cloned and analyzed. All of the promoters are TATA-less, lacking a CCAAT box but containing GC-rich boxes, characteristic of the house-keeping genes, although transcription of ganglioside synthase genes is subject to complex developmental and tissue-specific regulation. A set of cis-acting elements and transcription factors, including Sp1, AP2, and CREB, function in the proximal promoters. Negative-regulatory regions have also been defined in most of the promoters. We present here an overview of these genes and their transcriptional regulation. PMID- 18393826 TI - Aging and remodeling during healing of the wounded heart: current therapies and novel drug targets. AB - Aging has become a major health care problem and socio-economic burden worldwide. Myocardial infarction (MI) is the major killer worldwide and coronary reperfusion is the major form of acute post-MI therapy. The aging population is increasing, and with it, morbidity and mortality due to impaired healing after ST-segment elevation MI (STEMI) and its consequences. Optimal healing of the wounded heart is critical for preservation of structural and functional integrity of the pumping chambers, survival, and a favorable outcome irrespective of age. Although STEMI is more prevalent in the elderly and impaired healing during aging may promote adverse remodeling and thereby jeopardize outcome, there is an information gap on post-STEMI healing and its therapy in the elderly. Current therapies during post-STEMI healing are aimed primarily at the <65 age-group and preclinical studies tend to test drugs in mostly young animals. Therapies over the last decade have improved post-MI survival mainly in patients aged < 65 years. Novel healing-specific proteins may provide potential targets for improving healing and limiting adverse remodeling of the post-STEMI heart in the elderly, thereby improving outcome. PMID- 18393827 TI - Cellular and molecular mechanisms involved in the action of vitamin D analogs targeting vitiligo depigmentation. AB - The active metabolite of vitamin D3 - 1,25-(OH)2D3 - exerts most of its physiological and pharmacological actions through its nuclear receptor (VDR), regulating the transcriptional machinery of a variety of cell types. Basic research motivated by the detection of VDR in numerous target cells, has indicated potential therapeutic applications of VDR ligands in osteoporosis, cancer, secondary hyperparathyroidism and autoimmune diseases such as psoriasis, systemic lupus erythematosus, rheumatoid arthritis, type 1 diabetes and multiple sclerosis. In recent years vitamin D analogs, particularly calcipotriol and tacalcitol, have been used as topical therapeutic agents in vitiligo, an autoimmune pigmentary disorder characterized by aberrant loss of functional melanocytes from involved epidermis. The presence of cytotoxic T cells targeting melanocyte antigens and imbalance of the cytokine network were described as characteristics of the disease, eventually leading to melanocyte damage and death. Vitamin D ligands are designed to target the local immune response in vitiligo, acting on specific T cell activation, mainly by inhibiting the transition of T cells from early to late G1 phase and by inhibiting the expression of several pro-inflammatory cytokines genes, such as those encoding tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma). Vitamin D(3) compounds are known to influence melanocyte maturation and differentiation and also to up-regulate melanogenesis through pathways activated by specific ligand receptors, such as endothelin receptor and c-kit. In this review we summarize the complex pathogenetic rationale of vitamin D analogs in vitiligo depigmentation. Understanding the cellular and molecular mechanisms through which vitamin D targets the epidermal melanin unit is of great interest for identification of new effective therapeutic combination(s) that might induce repigmentation in vitiligo. PMID- 18393828 TI - Viral vectors in cancer immunotherapy: which vector for which strategy? AB - Gene therapy involves the transfer of genetic information to a target cell to facilitate the production of therapeutic proteins and is now a realistic prospect as a cancer treatment. Gene transfer may be achieved through the use of both viral and non-viral delivery methods and the role of this method in the gene therapy of cancer has been demonstrated. Viruses represent an attractive vehicle for cancer gene therapy due to their high efficiency of gene delivery. Many viruses can mediate long term gene expression, while some are also capable of infecting both dividing and non-dividing cells. Given the broadly differing capabilities of various viral vectors, it is imperative that the functionality of the virus meets the requirements of the specific treatment. A number of immunogene therapy strategies have been undertaken, utilising a range of viral vectors, and studies carried out in animal models and patients have demonstrated the therapeutic potential of viral vectors to carry genes to cancer cells and induce anti-tumour immune responses. This review critically discusses the advances in the viral vector mediated delivery of immunostimulatory molecules directly to tumour cells, the use of viral vectors to modify tumour cells, the creation of whole cell vaccines and the direct delivery of tumour antigens in animal models and clinical trials, specifically in the context of the suitability of vector types for specific strategies. PMID- 18393829 TI - Virus-based gene transfer approaches and adipose tissue biology. AB - The status of adipose tissue changes rapidly. From a simple filler tissue, it successively acquires the status of metabolic active tissue, endocrine tissue, plastic tissue, and finally that of a large reservoir of cells suitable for cell therapy and regenerative medicine. All throughout this story, our knowledge has been largely dependent on genetic tools and gene transfer. Now, the time has come where gene transfer in adipose derived cells can be envisioned, not only for understanding the role or importance of one gene, but also to engineer adipose derived cells for the purpose of therapy by delivering secreted products. In this paper, after a brief overview of adipose tissues, a large part will be devoted to the use of virus-based gene transfer in transducing adipose tissue and cells which reside therein. We also critically review the use of adipose "specific" promoters and the applications already described in the literature. PMID- 18393830 TI - Virotherapy as an approach against cancer stem cells. AB - It has been hypothesized that cancers originate from a small population of cells with stem cell-like characteristics, including self-renewal and pluripotency. Such tumor-initiating cells, also referred to as cancer stem cells, are thought to account for relapses following seemingly successful treatments, because their slow turnover and capacity for expelling anti-tumor drugs leaves them untouched by conventional treatment regimens. Targeting of cancer stem cells might be key for improving survival and producing cures in patients with metastatic tumors. Viruses enter cells though infection and might therefore not be sensitive to stem cell resistance mechanisms. During the last decades, oncolytic adenoviruses have been shown to effectively kill cancer cells, by seizing control of their DNA replication machinery and utilizing it for the production of new virions, ultimately resulting in the rupture of the cell. Human safety data in cancer trials has been excellent even when the dose of administered adenovirus has been high. Future approaches include additional modifications of the adenoviral genome that prime them to attack cancer stem cells specifically, utilizing linage specific cell surface markers, dysfunctional stem cell signaling pathways or up regulated oncogenic genes. However, already existing oncolytic adenoviruses have displayed potential to efficiently kill not only differentiated cancer cells, but also tumor-initiating stem cells. Here, we review the current literature that supports the existence of cancer stem cells and discuss the potential of virotherapy for killing tumor-initiating cells. PMID- 18393831 TI - The poxvirus vectors MVA and NYVAC as gene delivery systems for vaccination against infectious diseases and cancer. AB - Recombinants based on poxviruses have been used extensively as gene delivery systems to study many biological functions of foreign genes and as vaccines against many pathogens, particularly in the veterinary field. Based on safety record, efficient expression and ability to trigger specific immune responses, two of the most promising poxvirus vectors for human use are the attenuated modified vaccinia virus Ankara (MVA) and the Copenhagen derived NYVAC strains. Because of the scientific and clinical interest in these two vectors, here we review their biological characteristics, with emphasis on virus-host cell interactions, viral immunomodulators, gene expression profiling, virus distribution in animals, and application as vaccines against different pathogens and tumors. PMID- 18393832 TI - Gene transfer to sperm and testis: future prospects of gene therapy for male infertility. AB - Male infertility has been considered a major contributory factor to infertility. The causes of spermatogenetic failure found in most cases of male infertility remain largely idiopathic. Unfortunately, there is no effective treatment to improve spermatogenesis for idiopathic male infertility patients. Intracytoplasmic sperm injection (ICSI) is the current treatment of choice for severe male infertility and has brought the joy of childbearing to couples for whom it was previously impossible; however, several problems exist with this treatment. In addition, if there are no spermatozoa in the testis of these patients, they do not have paternity potential even if ICSI is conducted. Ultimately, fertilization is better in vivo than in vitro. Recently, on the other hand, gene transfer to sperm and testis has been developed to find more effective and simple methods to obtain transgenic animals. This technique has the potential to be the most useful approach for the future treatment of male infertility. In this review, we will give an overview of the recent advanced technique of gene transfer to sperm and testis, and discuss the future prospects of gene therapy for the treatment of male infertility. In conclusion, although more investigations on the mechanism of spermatogenesis and male infertility and the establishment of techniques for more efficient and safer gene transfer to the sperm and testis will be needed, gene therapy will enable a revolutionary advance for reproductive treatment and provide great benefit for patients with male infertility in the future. PMID- 18393833 TI - Human hematopoietic stem cells in gene therapy: pre-clinical and clinical issues. AB - Hematopoietic stem and progenitor cells (HSC) have been widely used in allogeneic transplant procedures, therefore their intrinsic characteristics, the biology of their niche in the bone marrow, and the mobilization and homing processes have been extensively investigated. With the development of gene therapy strategies, new therapeutic options based on autologous HSC have become available which may reduce the morbidity and mortality associated to allogeneic transplantation, but require an ex vivo manipulation of the cells to be corrected before re-infusion. For the success of these approaches it is necessary to optimize culture conditions in order to achieve efficient cell transduction while preserving the biological properties of the stem cells. We review here the factors critical for achieving efficient HSC transduction and maintenance of HSC stemness and homing capacity upon ex vivo culture. When HSC gene therapy is used in genetic disorders, permanent integration of therapeutic genes into the chromosomes of affected cells is needed. Indeed, by use of integrating vectors, such as retroviruses, gene therapy has met significant success in immunodeficiency syndromes characterized by a selective advantage of the transduced cells. However, retroviral integration can take place in stem cells at a variety of chromosomal sites, and examples have been reported of integration of therapeutic vectors causing cancer in patients. The clinical benefit arising from the long term correction of the genetic defect, due to vector integration into the HSC genome, and the adverse consequences of these events are also here discussed, together with the new and challenging perspectives of HSC gene therapy. PMID- 18393841 TI - Testing of compounds in models of pulmonary emphysema. AB - There is a pressing need for the development of new therapies for emphysema, particularly as no existing treatment has been shown to reduce disease progression. Compounds with a potential activity against the pathological mechanisms postulated to play a role in the development and progression of emphysema should be tested in vivo in animal models of this disease. The choice of the model is of capital importance. While models of elastase-induced emphysema are relatively easy to execute, require low personnel capacity and provide fast results, they also have a limited clinical relevance. On the other hand, models of chronic smoke exposure are time-consuming, expensive and require high personnel capacity but have a high clinical relevance. Presently, mainly two pharmacological approaches are being considered and investigated in experimental studies. The first approach consists of pharmacological interventions designed to slow down the rate at which alveolar wall is lost in emphysema. In this approach we find anti-inflammatory agents, protease inhibitors and antioxidants. The attempt to reduce lung inflammatory cell infiltration is most appealing since such an effect would also reduce the lung burden of both proteases and oxidants. The second approach is an attempt to reverse the process of alveolar loss by inducing alveolar growth. To our knowledge here only the effects of retinoids and/or retinoid receptor agonists have been investigated. This report presents a selected review of the literature of animal studies using these pharmacological approaches. PMID- 18393842 TI - Conventional and gene therapy strategies for the treatment of brain tumors. AB - There are several types of primary tumors of the central nervous system (CNS), and almost half of them are gliomas. In particular, glioblastoma multiforme (GBM) is very aggressive and infiltrates into the CNS parenchyma. Despite intense clinical efforts, the prognosis of patients with this type of tumors remains very poor, and has not improved in decades, with a median survival of approximately one year after diagnosis. Current treatments include surgical resection, radiotherapy and chemotherapy. However, our knowledge regarding the genetic basis, as well as the molecular signaling pathways involved in the origin and progression of the tumors has increased significantly in the last few years, thus allowing the generation of new chemotherapeutic agents that are used together with sophisticated surgical and radiation techniques. Nevertheless, new approaches are necessary to develop effective treatments for these tumors. One of these novel strategies is gene therapy which is particularly well suited to treat gliomas. In this review we will discuss current therapeutic approaches, as well as critically analyzing gene therapy methods, the use of diverse viral and non viral vectors, different genes and strategies to treat gliomas, from experimental models to clinical applications. PMID- 18393843 TI - Entry of oximes into the brain: a review. AB - The passage of hydrophilic drugs, such as oxime acetylcholinesterase reactivators, into the central nervous system is restricted by the blood-brain and the blood-cerebrospinal fluid barriers. The present review summarizes morphological and functional properties of the blood-brain barrier, blood cerebrospinal fluid barrier and cerebrospinal fluid-brain interface and reviews the existing data on brain entry of oximes. Due to the virtual absence of transcytosis, lack of fenestrations and unique properties of tight junctions in brain endothelial cells, the blood-brain barrier only allows free diffusion of small lipophilic molecules. Various carriers transport hydrophilic compounds and extrude potentially toxic xenobiotics. The blood-cerebrospinal fluid barrier is formed by the choroid plexus epithelium, whose tight junctions are more permeable than those of brain endothelial cells. The major function of plexus epithelium cells is active transport of ions for the production of the cerebrospinal fluid. The cerebrospinal fluid-brain interface is not a biological barrier and allows free diffusion. However, in contrast to passage via the blood-brain barrier or the blood-cerebrospinal fluid barrier, direct penetration from the cerebrospinal fluid into the brain is very slow, since much longer distances have to be covered. A bulk flow of brain interstitial fluid and cerebrospinal fluid speeds up exchange between these two fluid compartments. Oximes, by reactivating acetylcholinesterase, are important adjunct therapeutics in organophosphate poisoning. They are very hydrophilic and therefore cannot diffuse freely into the central nervous system. Changes in brain acetylcholinesterase activity, oxime concentration and some biological effects elicited by oxime administration in the periphery indicate, however, that oximes can gain access to the brain to a certain degree, probably by carrier-mediated transport, reaching in the brain about 4-10% of their respective plasma levels. The clinical relevance of this effect is hotly debated. Possible strategies to improve brain penetration of oximes are discussed. PMID- 18393844 TI - New tubulin polymerization inhibitor derived from thalidomide: implications for anti-myeloma therapy. AB - Despite the conventional and high-dose chemotherapy with hematopoietic stem cell transplantation, multiple myeloma eventually relapses, resulting in an incurable hematological malignancy. Therefore, novel therapeutic approaches in clinical settings are desired. Recently, thalidomide was introduced for the treatment of myeloma, and many clinical trials have since confirmed its efficacy in patients with relapsed/refractory or newly diagnosed multiple myeloma. Multiple mechanisms have been proposed to explain thalidomide's anti-myeloma activity. However, the precise mechanism underlying this activity remains unclear, because thalidomide rapidly undergoes spontaneous, nonenzymatic, hydrolytic cleavage to numerous metabolites in vivo. To elucidate the exact anti-myeloma mechanism of thalidomide in vivo, we have performed structural development studies of thalidomide, and obtained various analogs with specific molecular properties. Among these derivatives, we found that a new thalidomide analog, 2-(2,6-diisopropylphenyl)-5 hydroxy-1H-isoindole-1,3-dione (5HPP-33), has the most potent anti-myeloma effect with tubulin polymerization inhibiting activity. 5HPP-33 directly inhibited the growth and survival of various myeloma cells in a dose-dependent manner with IC(50) of 1-10 microM. In contrast, thalidomide itself did not inhibit RPMI8226 cell growth. A tubulin polymerization assay using microtubule protein from porcine brain revealed that 5HPP-33 had potent tubulin polymerization inhibiting activity with IC(50) of 8.1 microM, comparable to that of rhizoxin, a known tubulin polymerization inhibitor. Moreover, its activity was more potent than that of a known thalidomide metabolite, 5-hydroxythalidomide. Our data suggest that 5HPP-33 is a promising candidate as a therapeutic agent for multiple myeloma. In addition, the results suggest that thalidomide's tubulin polymerization inhibiting activity might be the mechanism underlying the induction of apoptosis in myeloma cells. PMID- 18393845 TI - Amyloid-related biomarkers for Alzheimer's disease. AB - Alzheimer's disease (AD) is an age-related disorder that causes brain damage resulting in progressive cognitive impairment and death. Three decades of progress have given us a detailed understanding of the underlying molecular mechanisms. Over the past 10 years, this knowledge has translated into a range of targets for therapy, the most promising of which is amyloid beta (Abeta). An imbalance between the production and clearance of Abeta is thought by many to represent the earliest event in the pathogenesis of AD. Abeta is known to be subject to oligomerisation, a process that increases its synaptotoxicity. The oligomers may aggregate further to proto-fibrils and fibrils, eventually forming senile plaques, the neuropathological hallmark of AD. In this article we review the key aspects of Abeta as a biomarker for AD, including its pathogenicity, the diagnostic performance of different Abeta assays in different settings, and the potential usefulness of Abeta as a surrogate marker for treatment efficacy in clinical trials of novel Abeta-targeting drugs. PMID- 18393846 TI - The other side of the opioid story: modulation of cell growth and survival signaling. AB - Opioids have been used as pain control medications for thousands of years. Opioids are highly effective analgesics clinically available for controlling moderate and severe pain. Recent genetic knockout and knockin studies have definitively demonstrated that the analgesic effect is mediated through opioid receptors. In addition to their analgesic effect, opioids also have the potential to develop tolerance and physical dependence. Moreover, opioids can modulate cell proliferation and survival. Attempts to design better opioid drugs to eliminate or diminish these undesirable effects for clinical benefits have achieved limited success. In recent years, investigation of the effects of opioid-mediated cell proliferation and survival has been very active, resulting in many publications. However, the molecular targets of such non-analgesic effects are complex. Several important pathways that control cell proliferation, survival, and apoptosis have been reported to be associated with the non-analgesic effects, which may be mediated through both opioid receptor signaling and other non-opioid receptor molecular entity-mediated signaling. This review tries to bring the attention of the medicinal chemistry community to new developments and advances in the research areas of opioid-mediated cell proliferation and survival. Further investigation of the molecular mechanism of these non-analgesic opioid effects may eventually yield useful information such as new drug targets, which may be explored to benefit for clinical treatments such as targeted cancer therapy, cancer pain management, regeneration of neurons, and recovery from drug addiction. PMID- 18393847 TI - Lessons from viruses: controlling the function of transmembrane proteins by interfering transmembrane helices. AB - Viral proteins sometimes interfere with human transmembrane receptors to gain access into a cell or they use transmembrane domains to interfere with cellular signal cascades in human cells. Such interference can lead to a deregulation of tightly regulated processes and eventually to different forms of cancer. There is still little knowledge about how proteins act and interact in biological membranes but the membrane environment restricts the fold and composition of membrane proteins when compared to water soluble proteins. These restrictions and a sometimes related functional principle of different viral transmembrane proteins for gaining access to a host cell or to intervene with cellular processes may offer a great opportunity to interfere with those processes in a simplified manner. A close collaboration of various disciplines may result in the development of drugs that specifically target membranes and interfere with viral transmembrane domains or even regulate the function of cellular membrane proteins. In this review we describe the function of interactions between human and viral membrane proteins in a cellular membrane, and perspectives to intervene with those processes are discussed. PMID- 18393848 TI - Hyaluronic acid: targeting immune modulatory components of the extracellular matrix in atherosclerosis. AB - PURPOSE OF REVIEW: Atherosclerosis is a chronic inflammatory disease of the vessel wall. Although it has become common knowledge that hyaluronic acid (HA), an important component of the extracellular matrix, is strongly involved in atherosclerotic disease development it has only recently become evident that HA, instead of being a static matrix polymer, is an active modulator of proliferation and inflammation of the atherosclerotic plaque. This review discusses the regulatory capacity of HA in atherosclerotic lesions and its effects on plaque stability. RECENT FINDINGS: The mechanisms by which HA might alter plaque stability are diverse. It regulates cellular migration and proliferation, lipid accumulation, and intraplaque angiogenesis. Smooth muscle cell migration is enhanced upon accumulation of HA, potentially stabilizing atherosclerotic plaques. On the other hand, HA is an important ligand for CD44, which stimulates inflammatory cell recruitment to lesions, leukocyte migration and cell proliferation in atherosclerotic plaques. Furthermore, HA forms complexes with low density lipoproteins, and uptake of these complexes by macrophages is increased compared to native LDL, indicating a more detrimental effect on atherosclerosis. The dynamic functional role of HA might be based on the functional difference between short and larger size fragments of this polymer, with either an inflammatory or an anti-inflammatory besides a pro-mitogenic and anti-mitogenic effect. Low molecular weight HA has been shown to be pro angiogenic, whereas high molecular weight HA has an anti-angiogenic effect. The cause of these differential actions might be that HA synthesis is regulated by three different genes: HAS1, 2 and 3 leading to different size HA products. These genes are specifically expressed under certain conditions, e.g. HAS1 and HAS3 are selectively induced in inflammation, suggesting an important role of their products in this process. SUMMARY: Hyaluronic acid is an active regulatory component of atherosclerotic lesions. Further studies are warranted to gain more insight into the mechanisms which decide on the role of HA in atherosclerosis and plaque stability. PMID- 18393849 TI - Novel molecules targeting dyslipidemia and atherosclerosis. AB - Atherosclerotic cardiovascular disease (CVD) is the leading cause of morbidity and mortality worldwide. Dyslipidemia is one of the main risk factors leading to atherosclerosis. Moreover, there is evidence for a role of oxidation in linking lipids and inflammation to development and progression of atherosclerosis. Current therapeutic approaches with lipid-lowering agents, such as statins, fail to protect more than half of patients from cardiovascular events. Therefore, there is a need for additional and alternative treatment options. There are several novel molecules undergoing preclinical or clinical development for the treatment of dyslipidemia or against distinct pathways which contribute to the development of atherosclerosis. Novel squalene synthase inhibitors with significant cholesterol-lowering and antiatherosclerotic properties are under development. Targeting the production of apolipoprotein B-100 with an antisense oligonucleotide is another interesting approach for lowering low density lipoprotein(LDL)-cholesterol levels. Raising high density lipoprotein(HDL) cholesterol levels or improving its antiatherosclerotic properties constitute additional attractive targets for protection against CVD. Such compounds include the cholesteryl ester transfer protein inhibitors, HDL-derived proteins, and mimetic peptides/lipids. Direct targeting of atherosclerosis remains a challenge. Molecules against oxidation and/or inflammation could be beneficial in reducing atherosclerosis. Other targets involved in distinct pathways of atherosclerosis include the lipoprotein-associated phospholipase A(2), 5-lipoxygenase-activating protein, acyl-CoA:cholesterol acyltransferase, chemokine receptors, and protein kinases. In conclusion, there are several promising novel therapeutic approaches for dyslipidemia and atherosclerosis under development which are expected to be of great benefit for patients at risk of CVD. PMID- 18393850 TI - Biopharmaceutics, pharmacokinetics and pharmacodynamics of antituberculosis drugs. AB - Tuberculosis (TB) is the leading cause of mortality due to a single infectious agent. The currently used combination drug regimens produce cure rates that exceed 95%, given good patient adherence during the multiple months treatment period. However the recent surge in HIV infections and the synergy between HIV and TB as well as the emergence of resistance resulted in an unforeseen increase in the number of TB cases, including multi-drug resistant (MDR) and extensively drug resistant (XDR) forms of TB. Consequently, there is an urgent need to develop novel, fast acting antituberculosis drugs with high potency that can provide treatment options for all forms of TB. It is well known that the current TB drugs exhibit differences in their in vivo activity profile and these differences are largely determined by their pharmacodynamics (PD), i.e. intrinsic antibacterial activity, biopharmaceutical properties such as solubility and permeability, and pharmacokinetic (PK) properties such as drug exposure, tissue distribution, and protein binding. An understanding of the relationships among these properties is considered key for a rational use of antituberculosis therapeutics. The current review provides a comprehensive summary of physicochemical/biopharmaceutical, PK, and PD properties of currently used antituberculosis drugs and novel agents under development. Also, a brief review of PK/PD parameters of current TB drugs is given and properties of a desirable TB drug target and drug molecule are outlined. The information provided herewith may be useful in the optimization of biopharmaceutical and PK/PD characteristics in the development of novel TB therapeutics and in the design of optimal treatment regimens. PMID- 18393851 TI - Microarray gene expression profiling in meningiomas and schwannomas. AB - Microarray gene expression profiling is a high throughput system recently used in basic and applied research. It provides a large amount of data -at molecular level- that once acquired, must be functionally integrated in order to find common patterns within a defined group of biological samples. In addition to identification of differentially expressed genes and the establishment of gene regulation patterns, microarrays may also allow us to discover new tumor markers that could have a great impact on the improvement of clinical practice and therapeutics for cancer. The classification method used for cancer is currently based on the morphological characteristics of the biological samples. The information obtained with this method is limited, omitting many important tumor characteristics like the proliferation rate, the capacity of invasion and metastases, as well as the possible development of mechanisms of cellular resistance to treatment. Microarrays can be used in combination with conventional diagnostics as a helpful complement. In this review we focus on how this technology has contributed to our knowledge of the molecular pathogenesis of meningiomas and schwannomas, its potential role as a useful tool for tumor classification and its application in clinical practice. PMID- 18393852 TI - Oxidatively damaged DNA repair defect in cockayne syndrome and its complementation by heterologous repair proteins. AB - Cockayne syndrome (complementation groups A and B) is a rare autosomal recessive DNA repair disorder characterized by photosensitive skin and severely impaired physical and intellectual development. The Cockayne syndrome A and B proteins intervene in the repair of DNA modifications that block the RNA polymerase in transcribed DNA sequences (transcription-coupled repair). Recent results suggest that they also have a more general role in the repair of oxidative DNA base modifications. Although the phenotypical consequences of defective repair of oxidatively damaged DNA in Cockayne syndrome are not determined, accumulation of oxidized lesions might contribute to delay the physical and intellectual development of these patients. To conceive new therapeutic strategies for this syndrome, we are investigating whether the oxidatively damaged DNA repair defect in Cockayne syndrome might be complemented by heterologous repair proteins, such as the Escherichia coli formamidopyrimidine-DNA glycosylase and endonuclease III. The complementation studies may shed light on the important lesions for the Cockayne syndrome phenotype and offer new tools for future therapies aimed at counteracting the consequences of oxidatively damaged DNA accumulation. PMID- 18393853 TI - Current developments in the synthesis and biological activity of aza-C nucleosides: immucillins and related compounds. AB - This review will describe the recent advances in the field of aza-C-nucleosides with a particular emphasis on immucillins and related compounds. The review will cover both chemical and biological aspects concerning their preparation and/or occurrence in Nature as well as their biological properties which include glycosidase, glycosyl transferase, and nucleoside hydrolase and phosphorylase inhibition, among others. These enzymatic inhibitory properties are the basis for the potential use of the title compounds in viral and parasitic infections, cancer and genetic disorders. PMID- 18393854 TI - Granulocyte colony-stimulating factor (G-CSF) for cardio- and cerebrovascular regenerative applications. AB - The cytokine granulocyte colony-stimulating factor (G-CSF) is produced by numerous cell types including immune and endothelial cells. G-CSF binding to its receptor G-CSF-R which belongs to the cytokine receptor type I family depends on the interaction of alpha-helical motifs of the former and two fibronectin type III as well as an immunoglobulin-like domain of the latter. It activates several signalling transduction pathways including PI3K/Akt, Jak/Stat and MAP kinase, thereby promoting survival, proliferation, differentiation and mobilisation of haematopoietic stem and progenitor cells. Accordingly, recombinant human (rh)G CSF has been extensively used in clinical haematology and oncology to enable bone marrow transplantation or to treat chemotherapy-associated neutropenia. Using animal models it has been recently shown that G-CSF, alone or in combination with other cytokines such as stem cell factor (SCF), causes an accumulation of bone marrow-derived cells in the infarcted heart which, however, do not differentiate into cardiac cells. Nevertheless, since beneficial effects on structural and functional properties were observed in animal models of cardiac, brain and hindlimb ischaemia other mechanisms of G-CSF action must be operative. Recent evidence suggests paracrine effects mediated by the immigrated bone marrow derived cells and/or direct effects of the cytokine on resident G-CSF-R expressing cells. In both cases these may include promotion of cellular survival, proliferation and differentiation. First clinical studies in patients with myocardial infarction, heart failure and stroke have been accomplished and are reviewed in this paper. PMID- 18393855 TI - ANTI-ADHESION evolves to a promising therapeutic concept in oncology. AB - Adhesion is a hallmark of haematological and solid cancer cells. All five classes of cell adhesion molecules (CAM) - integrins, cadherins, immunoglobulin-like CAMs, selectins and CD44s - are characteristically dysregulated in human cancer. Adhesion enables and promotes cancer-defining biological processes like growth, survival, migration, extravasation, homing, and metastasis. Furthermore, cell adhesion mediates drug resistance (CAM-DR) in multiple myeloma, malignant lymphoma, acute and chronic leukaemias, as well as in pancreatic cancer, neuroblastoma, small cell and non-small cell lung cancer, mesothelioma, colorectal carcinoma, and breast cancer. Cell adhesion protects from death by radiation, genotoxic chemotherapy, or targeted pathway inhibitors. Adhesion molecules are overexpressed on drug resistant cells (e.g. multiple myeloma or prostate cancer). Very recently, several cell adhesion mediated survival pathways have been elucidated, with key mediators being LFA-1, VLA-4, FAK, ILK, Src, PI3K, Akt, Ras, MEK, Erk, HMG-CoA reductase, Rho, Rho kinase, PKC, and NFkB. Because the surface and the intracellular targets are now known and because specific compounds are becoming increasingly available, first clinical trials regarding ANTI-ADHESION therapies are ongoing. However, in comparison to the comprehensive preclinical and clinical knowledge about CAMs, the number of drugs developed thusfar is quite low. ANTI-ADHESION strategies include targeting of surface antigens, inhibition of cell adhesion associated pathways, inhibition of CAM-DR, and targeted drug delivery. As ANTI-ADHESION is based on general characteristics of cancer cells independent of specific disease entities or treatment modalities, it may become a successful, low-toxic and broadly applicable concept in cancer treatment. PMID- 18393856 TI - Anti-infective and anti-tumor agents based on the depletion of immune suppressive effects. AB - Aggressive immunity characterized by the motion of cytotoxic T lymphocytes (CTLs), T helper (Th) 1 cells, and natural killer (NK) cells is the first line of defense against intracellular microorganism invasion and tumor formation. In patients with infectious diseases and tumors, aggressive immunity is often attenuated by immune suppressive effects provided by regulatory T (Treg) cells including CD4(+) CD25(+) forkhead-box (fox) p3(+) T cells, T regulatory (Tr) 1, Th3, and a subpopulation of gammadelta-type of T cell receptor-expressing T (gammadelta T) cells. It has been demonstrated that Treg cells down-regulate aggressive immunity by direct cell interactions and suppressive cytokines (e.g., interleukin (IL)-10, transforming growth factor (TGF)-beta). Today, instead of synthesizing chemical agents with serious side effects, protein agents, catalytic oligonucleotides, and natural medicines involved in the elimination of Treg cell mediated suppressive responses for the restoration of aggressive immunity are expected to be alternatives as a mild clinical remedy against microorganism invasion and tumors. PMID- 18393857 TI - Arbidol: a broad-spectrum antiviral compound that blocks viral fusion. AB - Arbidol (ARB; ethyl-6-bromo-4-[(dimethylamino)methyl]-5-hydroxy-1-methyl-2 [(phenylthio)methyl]-indole-3-carboxylate hydrochloride monohydrate), is a Russian-made potent broad-spectrum antiviral with demonstrated activity against a number of enveloped and non-enveloped viruses. ARB is well known in Russia and China, although to a lesser extent in western countries. Unlike other broad spectrum antivirals, ARB has an established molecular mechanism of action against influenza A and B viruses, which is different from that of available influenza antivirals, and a more recently established mechanism of inhibition of hepatitis C virus (HCV). For both viral infections the anti-viral mechanism involves ARB inhibition of virus-mediated fusion with target membrane and a resulting block of virus entry into target cells. However, ARB inhibition of fusion exploits different ARB modalities in case of influenza viruses or HCV. This review aims to summarize the available evidence of ARB effects against different groups of viruses, also, to compare various aspects of ARB anti-fusion mechanisms against influenza virus and HCV (with reference to different stringency of pH-dependence of these two viral fusogens) and to discuss further prospects for ARB and its improved derivatives of the parent compounds. PMID- 18393858 TI - Non-steroidal targets in the diagnosis and treatment of endometriosis. AB - Endometriosis, a chronic gynecologic disease frequently resulting in chronic pelvic pain, severe dysmenorrhoea, and subfertility, is defined as the presence of endometrial tissue at extrauterine locations, most commonly on the peritoneum and ovaries. Conclusive diagnosis requires laparoscopic surgery followed by histological confirmation. The treatment options -at present- are limited to hormonal therapies and/or surgical ablation of the lesions, and are characterized by high recurrence rates, significant side-effects and limited duration of administration. The pathogenesis of endometriosis is still unclear and numerous immunological and inflammatory factors have been suggested to be involved in the development of the disease, including interleukin (IL)-1, IL-2, IL-6, IL-8, IL 12, tumour necrosis factor -alpha (TNF-alpha), regulated on activation, normal T Cell expressed and secreted (RANTES) and its receptor cognate chemokine receptor 1 (CCR1), peroxisome proliferator activated receptors (PPARs), matrix metalloproteinases (MMPs) and cyclooxygenase (COX). Another crucial mechanism in endometriosis is the vascularisation of the endometriotic lesions, with a key role for vascular endothelial growth factor (VEGF). Recently, protease activated receptors (PARs), mitogen-activated protein kinases (MAPKs) and tyrosine kinases have also been associated with the pathophysiology of endometriosis. The aim of this article is to discuss molecules that have recently been found to have connections with the pathogenesis of endometriosis, as potential targets to develop new methods for noninvasive diagnosis and for novel medical management of this disease. This review also critically addresses how these molecules can be tested in basic, preclinical and clinical research, the status of this research and the importance of potential side effects. PMID- 18393859 TI - Pharmacophore-based virtual screening. AB - Virtual screening (VS) is an important component of cheminformatics and molecular modeling. An abundance of structural information, indicated by both the ever increasing availability of 3-dimensional (3D) protein structures and the readiness of free conformational databases of commercially available compounds, such as ZINC, supplies a broad platform for VS. At the same time, new technology enables the implementation of more accurate and sophisticated pharmacophore models and the screening of millions of compounds within a manageable period. Therefore, VS is expected to play a more important role in future drug discovery efforts. This paper will examine and compare the advantages and disadvantages of VS against experimental high-throughput screening (HTS). It will also evaluate pharmacophore-based VS against docking-based VS. The strategies leading to successful pharmacophore-based VS are outlined, including how to enumerate a conformational database efficiently, how to select chemical features for a specific pharmacophore model, how to incorporate excluded volumes to enhance the geographical restrictions, and how to optimize a pharmacophore model. Successful examples of pharmacophore-based VS will be presented. PMID- 18393860 TI - Melanin-concentrating hormone receptor 1 antagonists: a new perspective for the pharmacologic treatment of obesity. AB - Obesity is a chronic disease characterized by the accumulation of excess adipose tissue associated with an increased risk of multiple morbidities and mortality. At the present time, only three drugs have been approved by the Food and Drug Administration (FDA) for the treatment of obesity. Agonists and antagonists of some of the substances implicated in the regulation of energy homeostasis represent opportunities for anti-obesity drug development. The most promising targets are alpha-melanocyte stimulating hormone (alpha-MSH) receptors, cannabinoid receptors, the 5-hydroxytryptamine (5-HT) receptors and melanin concentrating hormone (MCH) receptors. MCH receptors could be major potential targets for the treatment of obesity. Many pharmaceutical companies have described MCH-R1 antagonists that have appeared over the past year. Recently, two compounds went into phase I clinical trials that evaluate MCH receptor antagonists as a new perspective for the pharmacologic treatment of obesity. In this review, structure-activity relationships (SAR) in the development of MCH-R1 antagonists are provided. PMID- 18393861 TI - Protein-protein interactions. PMID- 18393862 TI - Predicting 3D structures of protein-protein complexes. AB - The protein-protein docking problem is one of the focal points of activity in computational structural biology. Adequate computational techniques for structural modeling of protein interactions are important because of the growing number of known protein structures, particularly in the context of structural genomics. The protein docking methodology offers tools for fundamental studies of protein interactions and provides structural basis for drug design. The paper presents a critical review of the existing protein-protein docking approaches in view of the fundamental principles of protein recognition. PMID- 18393863 TI - Characterization and prediction of protein interfaces to infer protein-protein interaction networks. AB - Complex protein-protein interaction networks govern biological processes in cells. Protein interfaces are the sites where proteins physically interact. Identification and characterization of protein interfaces will lead to understanding how proteins interact with each other and how they are involved in protein-protein interaction networks. What makes a given interface bind to different proteins; how similar/different the interactions in proteins are some key questions to be answered. Enormous amount of protein structures and experimental protein-protein interactions data necessitate advanced computational methods for analyzing and inferring new knowledge. Interface prediction methods use a wide range of sequence, structural and physico-chemical characteristics that distinguish interface residues from non-interface surface residues. Here, we present a review focusing on the characteristics of interfaces and the current status of interface prediction methods. PMID- 18393864 TI - Recognition-induced conformational changes in protein-protein docking. AB - The ability to predict the three-dimensional structure of a protein complex starting from the isolated binding partners is becoming increasingly relevant. As our understanding of the molecular mechanisms behind protein-protein binding improves, so do the docking methods, however, it remains a challenge to adequately predict the unbound to bound transition. Side-chain flexibility is routinely handled and most docking methods allow for a certain degree of backbone flexibility, but systems undergoing moderate to large conformational changes can at present not correctly be modeled. The docking community is therefore putting an increased effort in the treatment of protein flexibility. Here we present a survey of the existing computational techniques to model protein flexibility in the context of protein-protein docking. PMID- 18393865 TI - Molecular recognition and binding free energy calculations in drug development. AB - The functional capabilities of biological systems, such as enzyme catalysis, nutrient import, and cell signaling, depend crucially on specific molecular interactions. In addition, the effects of common drugs also act through a mechanism of binding to specific biomolecular targets. Models for the prediction of binding affinity are used in basic research to study the molecular basis of biological function as well as in applied research to study the development of new drugs. This review will address the biological importance of molecular recognition as well as its influence on the development of pharmaceuticals. Further, a broad overview of computational approaches used for the prediction of biological activity and specifically binding free energy will be presented. PMID- 18393866 TI - Calculating pH and salt dependence of protein-protein binding. AB - Ionic strength- (or salt-) effects on the protein-protein binding free energy has been included in many computational studies, while comparatively fewer computational studies have looked at the corresponding effect of pH. The pH dependence can be very complex if several groups change protonation state, while the ionic strength dependence usually scales as ln(I), and the main challenge is to predict the magnitude of the correlation. However, there is now very strong indication that pH effects due to binding induced changes in protonation states make a non-negligible contribution to the binding energy of most protein-protein complexes. This observation, together with more efficient pKa prediction methods and the emergence of constant pH molecular dynamics simulations to model the protonation-dependent structural changes will spark more experimental and theoretical work in pH effects on protein-protein binding. PMID- 18393867 TI - In silico-in vitro screening of protein-protein interactions: towards the next generation of therapeutics. AB - Protein-protein interactions (PPIs) have a pivotal role in many biological processes suggesting that targeting macromolecular complexes will open new avenues for the design of the next generation of therapeutics. A wide range of "in silico methods" can be used to facilitate the design of protein-protein modulators. Among these methods, virtual ligand screening, protein-protein docking, structural predictions and druggable pocket predictions have become established techniques for hit discovery and optimization. In this review, we first summarize some key data about protein-protein interfaces and introduce some recently reported computer methods pertaining to the field. URLs for several recent free packages or servers are also provided. Then, we discuss four studies aiming at developing PPI modulators through the combination of in silico and in vitro screening experiments. PMID- 18393868 TI - Approaches and resources for prediction of the effects of non-synonymous single nucleotide polymorphism on protein function and interactions. AB - Almost all (99.9%) nucleotide bases are exactly the same in all people, however, the remaining 0.1% account for about 1.4 million locations where single-base DNA differences/polymorphisms (SNPs) occur in humans. Some of these SNPs, called non synonymous SNPs (nsSNPs), result in a change of the amino acid sequences of the corresponding proteins affecting protein functions and interactions. This review summarizes the plausible mechanisms that nsSNPs may affect the normal cellular function. It outlines the approaches that have been developed in the past to predict the effects caused by nsSNPs with special emphasis on the methods that use structural information. The review provides systematic information on the available resources for predicting the effects of nsSNPs and includes a comprehensive list of existing SNP databases and their features. While nsSNPs resulting in amino acid substitution in the core of a protein may affect protein stability irreversibly, the effect of an nsSNP resulting to a mutation at the surface of a protein or at the interface of protein-protein complexes, could, in principle be, subject of drug therapy. The importance of understanding the effects caused by nsSNP mutations at the protein-protein and protein-DNA interfaces is outlined. PMID- 18393869 TI - Current trends in the treatment of supraventricular tachycardia in pediatric age. PMID- 18393870 TI - Supraventricular tachycardia in infants: epidemiology and clinical management. AB - Supraventricular tachycardias (SVTs) are observed in 0,1-0,4% of the paediatric population and represent an important clinical problem with related significant health and social issues. Most tachycardias are paroxysmal, being associated with sudden onset and termination, and only a relatively small number of them is permanent, namely chronic. Paroxysmal tachycardias, in addition, can be either sustained (lasting > 30 seconds) or non-sustained whenever their duration is less. Most SVTs are due to re-entry, and only atrial tachycardia and and junctional ectopic tachycardia are caused by enhanced automaticity. Atrial tachycardia, however, can also be due, although rarely, to re-entry or to triggered activity. A prompt recognition of these arrthmias in children by pediatric cardiologist is essential for a correct clinical managemet of the patients. In this review, the epidemiologic data regarding the SVTs in pediatric age are reported along with the description of the pathophysiological mechanisms and the analysis of electrocardiographic findings to be considered for a correct clinical diagnosis and a rational therapeutic approach to these arrhythmias. PMID- 18393871 TI - Characterization of supraventricular tachycardia in infants: clinical and instrumental diagnosis. AB - Supraventricular tachycardia (SVT) is the most common symptomatic arrhythmias in children. Re-entry tachycardias are the most common form, on the contrary automatic tachycardias are relatively rare. There are four types or re-entry: along anomalous pathway with bi-directional (Wolff-Parkinson-White) or unidirectional conduction, intranodal re-entry, intra-atrial re-entry that is common after surgical procedure, and finally the uncommon sinus node re-entry. Automatic tachycardias may be atrial or junctional. The different types of tachycardia have a different incidence according to the age: in the first year of age re-entry along anomalous pathway is the dominant form, while intranodal reentry becomes common during adolescence. The age at the beginning of tachycardia is important for long term prognosis. When SVT starts in the first months of life it disappears in 80% of cases within the first year of life; on the contrary, if tachycardia starts later spontaneous remission is detected in only 15%-20% of patients. In infancy heart failure is the more common presenting symptom, thereafter palpitations become the principal cause of recognition of SVT. Syncope is reported in about 8% of cases and in another 15% usually neonates and infants, the SVT has an occasional detection. Electrocardiogram (ecg) usually allows the precise diagnosis of various types of SVT, and every effort should be made to record ecg during tachycardia. The parameters that should be evaluated are: heart rate, P wave axis, PR and RP interval, and finally presence or absence of AV block. Short lasting episodes should be difficult to be recorded; in these cases cardio-call and trans-telephonic transmission represent useful techniques to obtain SVT demonstration. Patients with SVT require a complete evaluation with others diagnostic techniques: echocardiogram, Holter monitoring, stress test, that should be chosen according the type of tachycardia. Electrophysiologic evaluation is now rarely performed for diagnostic purpose; trans-esophageal atrial stimulation being less invasive than intracardiac evaluation is more extensively employed when diagnosis of SVT is uncertain. Transesophageal stimulation is useful in the following situations: 1) evaluation of patients with symptoms suggestive of paroxistic tachycardia but without ecg documentation, 2) to assess the mechanism responsible for re-entry tachycardia: macro re-entry versus intranodal re-entry 3) to evaluate characteristics of anomalous pathway with bi-directional conduction, and 4)to terminate re-entrant SVT. PMID- 18393872 TI - Supraventricular tachycardia in fetus: how can we treat ? AB - The normal fetal cardiac rhythm is characterized by a regular heart rate ranging between 100 and 160 -180 beats/min with a normal 1: 1 atrioventricular electromechanical relationship during each cardiac cycle. Fetal tachycardia occurring in approximately 0.5% of all pregnancies and it is an important cause of fetal morbidity and mortality. A fetal tachycardic heart is at risk for developing low cardiac output, hydrops and ultimately fetal death or significant neurological morbidity. Different conditions can play a role to determine the natural history of tachycardic fetus as gestational age, underlying pathophysiology of the arrhythmia, fetal heart rate, duration of the tachyarrhythmia, and presence or absence of cardiac dysfunction. Reliable diagnosis in utero of fetal arrhythmia is possible by ultrasound examination of the fetal heart. In fact pulsed wave Doppler guided by two-dimensional echocardiography provided important information on cardiac rhythm as it study the blood flow from different chambers. With the introduction of the latest myocardial deformation methodology, the fetal tachyarrhythmias can be diagnosed more accu notrately. Precise diagnosis of cardiac arrhythmias in the fetus is crucial for a managed therapeutic approach. The choice of management is correlated to many factors: gestational age, underlying pathophysiology of the arrhythmia, fetal heart rate, duration of the tachyarrhythmia, and presence or absence of cardiac dysfunction. A large review of fetal arrhythmias was been reported in our work. PMID- 18393873 TI - Infants and children with tachycardia: natural history and drug administration. AB - Tachyarrhythmias can occur at any age from the developing fetus through adulthood. However, in deference to adult-onset ischemic cardiac issues, abnormal heart rhythms occurring in the young are often due to developmental alterations of the cardiac conduction tissue, genetically-inherited changes of myocardial cellular ion membrane properties and both pre- and post-surgical repair of associated structural congenital heart anatomical defects. And different from adults, abnormal rhythms occurring in the young can spontaneously disappear with progressive patient growth. Both supra- and ventricular tachyarrhythmias occur in the young although atrial rhythm abnormalities far exceed those of the ventricle. In both, pharmacologic therapies to alter tissue conduction and refractoriness remain the mainstay for initial intervention in the infant and young child, reserving more invasive and potentially harmful ablation therapies for drug refractory cases. The purpose of the review is to present common and uncommon tachyarrhythmias which can occur in the fetus and throughout infancy. Emphasis will be placed on their electrocardiographic identification, recognition of any associated structural congenital heart defects and recommended pharmacologic management. Drug therapies will be divided according to mechanism of action and discussions of which particular agent is potentially best-suited to treat which specific tachyarrhythmia. A listing of current pharmacologic agents used in the young with appropriate dosages is included. PMID- 18393874 TI - Pharmacological therapy in children with atrioventricular reentry: which drug? AB - Atrioventricular reentrant tachycardia (AVRT) is the most common cause of supraventricular tachycardia in young children. In nearly 70% of cases, there is manifest preexcitation on electrocardiogram. In the rest, the accessory pathway is concealed. Drugs control AVRT by affecting conduction through the atrioventricular node (beta-blockers, digoxin, verapamil) or accessory pathway (flecainide, propafenone) or both (sotalol, amiodarone). Adenosine is the drug of choice in acute management of AVRT in hemodynamically stable children. In adenosine-resistant cases, intravenous flecainide, procainamide, esmolol, propafenone and amiodarone are other treatment options. Hypotension and bradycardia can occur during administration of these drugs. Verapamil may be used to treat AVRT using a concealed pathway. Verapamil should be avoided in infants and in patients with decreased cardiac function. In chronic management, catheter ablation is the preferred treatment in older children with frequent AVRT. In infants and small children, ablation is associated with higher risk, and pharmacologic management is recommended. Beta-blockers are the preferred first line drugs for chronic management. In patients with concealed accessory pathway, digoxin and calcium channel blockers are alternative options. Sotalol, flecainide, propafenone and amiodarone can be prescribed in resistant cases. Flecainide and propafenone should be avoided in children with structurally abnormal hearts because of a higher risk of proarrhythmia. The initiation of flecainide, propafenone and sotalol therapy is recommended in an inpatient setting to monitor for proarrhythmias. PMID- 18393875 TI - Sudden death and ventricular preexcitation: is it necessary to treat the asymptomatic patients? AB - Currently, asymptomatic ventricular preexcitation, which has been put at rest for many decades, remains a clinical challenge as there are no predictors of sudden death, which can be the first clinical presentation of the syndrome. Identification of risk factors for sudden death is important, considering the availability of a definitive treatment. Now, as radiofrequency catheter ablation of accessory pathways has reported success rates approaching 100 percent without major complications in many centers worldwide, it becomes unacceptable that even one asymptomatic individual with WPW will die or will experience life-threatening arrhythmic events. In our extensive experience a short anterograde refractory period of accessory pathways, inducibility of sustained tachyarrhythmias and the presence of multiple accessory pathways are the strongest predictors of life threatening arrhythmias and sudden death. Therefore, it is not yet justified that, after an incidental diagnosis of WPW syndrome has been made, no risk stratification by invasive testing is done. Subjects at high risk, particularly if young or adolescent, should be identified and then ablated in the same session as they can develop lethal arrhythmic events within a few years and this is our current practice. Recently, we sent a questionnaire to investigate clinical practices over a large number of centers around the world about asymptomatic ventricular preexcitation. A total of 100 replies were received and the results demonstrate that there is worldwide agreement in performing invasive electrophysiologic testing and prophylactic ablation in selected subjects. These findings provide strong evidence to revisit current guidelines, which appropriately in the absence of evidence had been conservative. PMID- 18393876 TI - Pharmacological therapy in children with nodal reentry tachycardia: when, how and how long to treat the affected patients. AB - Atrio-ventricular nodal reentrant tachycardia (AVNRT) is a rare supra-ventricular tachycardia (SVT) in children and becomes more frequent in adolescents. Most of children with an AVNRT have a healthy heart thus rarely experiencing severe symptoms. Because of haemodynamic instability or risk of complications, recurrences of SVT may require a chronic therapy. Interruption of dual atrio ventricular nodal physiology is the basic mechanism to terminate AVNRT. This may be achieved by using anti-arrhythmic drugs or through Radiofrequency catheter ablation (RF). We aim to review the literature on the use of anti-arrhythmic drugs for the management of AVNRT in children aged more than 1 year and discuss the recommended dosages and the duration of a long term therapy. In the absence of comparative trials of risks and benefits between pharmacological therapy and RF and because of a greater clinical experience with anti-arrhythmic drugs, these last but not the least continue to be first-line therapy in the management of most SVT in children. Trials on pharmacotherapy in children with SVT in general and AVNRT in particular are lacking, use of anti-arrhythmic drugs being extrapolated from adult literature. Although Adenosine is becoming more used since it is the safest and effective drug in the acute setting, Digoxin continue to be the drug of first choice. Beta-blockers and Class I anti-arrhythmic are the second choice drugs with Flecainide being the preferred anti-arrhythmic drug for treatment failures. Amiodarone is rarely used as a chronic therapy in resistant cases. With the new advances in the RF technology, this therapy is becoming more safe and effective for AVNRT in children. Therefore, additional well-designed controlled trials are needed to further evaluate the comparative efficacy of anti arrhythmic drugs in the management of AVNRT in children, as well as to evaluate dosing and toxicity in various age groups and determine the duration of a chronic therapy as compared to a potential RF. PMID- 18393877 TI - Pharmacological therapy in children with atrial fibrillation and atrial flutter. AB - Heart rhythm disorders in children are not different, on electrocardiographic trace, from heart rhythm disorders in adults with the exception of incidence which is different according to the age. Paticularly, atrial flutter (FlA) and fibrillation (FA) are very uncommon arrhythmias in the general pediatric population. Generally atrial fibrillation and atrial flutter, in our experience, is a temporary heart rhythm disturbance connected to specifical and resovable reasons with the exception of Fontain's surgical correction of congenital heart diseases or cardiopathies with dilatation of both atria. Presenting symptoms, symptom history (e.g., frequency, duration, and severity), risk assessment, previous response to alternative treatment options, convenience and patient preference for a specific treatment option, and costeffectiveness of a treatment option are among the many factors that should be considered. Treatment of atrial flutter and fibrillation in pediatric age involves several options: Pharmacological therapy, Transoesophageal atrial pacing (TEAP), Electrical cardioversion and Catheter ablation. In this review we evaluated the physiopathology, the clinical features and the current terapeutical strategies for these arrythmias in paediatric age. PMID- 18393878 TI - Written consent to use the drug in children: the problem of off-label drugs. AB - Cardiac arrhythmias in pediatric patients have different mechanisms and frequencies compared to adult patients. There are many physiological differences between children and adults that may affect the pharmacodynamic and pharmacokinetic of the antiarrhythmic drugs in pediatric population. Children, and specially breast feeding children, cannot be considered low weighted adults to select antiarrhythmic drug doses. Although radiofrequency ablation has experienced great technological advances, it is performed in selected pediatric patients. Therefore, the main therapeutic strategy is the use of antiarrhythmic drugs in children. The medical management of arrhythmias in pediatric patients is challenging and complex. There are few clinical guidelines. There is scarce and incomplete information about the efficacy and safety of antiarrhythmic drugs in pediatric population. Most of the doses and drug administration intervals are extrapolated from adult population and applied to children. Antiarrhythmic drug doses have been extensively studied in adult population. However, in pediatric population, there are very few clinical trials and the safety of these drugs is not well known. In general, dose regimens are based on small uncontrolled studies, extrapolation of drug doses from studies performed in the adult population or physician experience. As a consequence, there is a need for further studies to assess the most effective antiarrhythmic drug regimens in children reducing the risk of side effects. Evidence suggests that medical research in pediatric population is necessary and morally valuable. But investigators involved must take care of moral and ethical values, including the respect for the child-subject and his parents or legal representatives, and this respect compels them to consider the patient and family in the decision making process. The participation request and the informed consent must be obtained according to the competitions the patient exhibits, trying to anticipate information about benefits and possible damages derived from the investigation in an understandable language for him. In our opinion the pharmacologic clinical investigation of antiarrhythmic treatments in pediatrics is necessary. More clinical studies must be carried out under rigorous scientific rules that contemplate the particular ethical dilemmas this population faces. PMID- 18393879 TI - Collateral effects of antiarrhythmics in pediatric age. AB - Although there are numerous reports of antiarrhythmic use in children, controlled, comparison trials of antiarrhythmic agents in children are virtually nonexistent and most data are obtained from case series of children treated. Effective and safe pharmacological therapy requires that the physicians attempt to identify a drug with the most appropriate profile to attack the most vulnerable parameter of the mechanisms of the cardiac arrhythmia with the least pro arrhythmic/collateral effects. Digoxin in patients with Wolf-Parkinson -White syndrome, verapamil in infants and intravenous quinidine should be avoided as there is clear evidence that they can cause serious side effects. Collateral effects of other antiarrhythmic drugs are discussed in details in this review. Well-designed, controlled trials are needed to further evaluate the comparative efficacy of antiarrhythmics in children, as well as to evaluate dosing and toxicity in various age groups. PMID- 18393880 TI - Transcatheter ablation of supraventricular tachycardias in pediatric patients. AB - Ablation has become an important treatment for many pediatric patients with common supraventricular tachycardias (SVTs). Many multicenter studies have documented that radiofrequency (RF) catheter ablation is a safe and effective procedure for treatment of a large variety of SVTs in children and adults with a high success rate and minimal complications. Novel electrophysiology technologies such as electroanatomic mapping and sophisticated ablating catheters have improved success rates and decreased complications of transcatheter ablation. Moreover, within the last several years, a new energy source using cryoenergy has evolved as a safe and effective alternative for catheter ablation for arrhythmogenic substrates traditionally associated with increased risk when using RF ablation. In this review pediatric transcatheter ablation practice is analysed and discussed with reference to current clinical guidelines. PMID- 18393881 TI - Scope and limitations of the co-drug approach to topical drug delivery. AB - Many currently available drugs show unfavourable physicochemical properties for delivery into or across the skin and temporary chemical modulation of the penetrant is one option to achieve improved delivery properties. Pro-drugs are chemical derivatives of an active drug which is covalently bonded to an inactive pro-moiety in order to overcome pharmaceutical and pharmacokinetic barriers. A pro-drug relies upon conversion within the body to release the parent active drug (and pro-moiety) to elicit its pharmacological effect. The main drawback of this approach is that the pro-moiety is essentially an unwanted ballast which, when released, can lead to adverse effects. The term 'co-drug' refers to two or more therapeutic compounds active against the same disease bonded via a covalent chemical linkage and it is this approach which is reviewed for the first time in the current article. For topically applied co-drugs, each moiety is liberated in situ, either chemically or enzymatically, once the stratum corneum barrier has been overcome by the co-drug. Advantages include synergistic modulation of the disease process, enhancement of drug delivery and pharmacokinetic properties and the potential to enhance stability by masking of labile functional groups. The amount of published work on co-drugs is limited but the available data suggest the co-drug concept could provide a significant therapeutic improvement in dermatological diseases. However, the applicability of the co-drug approach is subject to strict limitations pertaining mainly to the availability of compatible moieties and physicochemical properties of the overall molecule. PMID- 18393882 TI - Histone deacetylase inhibitors: new hope for rheumatoid arthritis? AB - Histone deacetylase (HDAC) inhibitors are a new family of anti-cancer agents currently undergoing clinical investigations for various oncology indications. Their anti-inflammatory activities had been well documented and they appear to be potential therapeutic strategies for various inflammatory diseases. In this review, the anti-inflammatory activities of HDAC inhibitors with emphasis on their potential applications in rheumatoid arthritis (RA) will be summarized. The possible anti-rheumatic mechanisms, including growth arrest in rheumatoid arthritis synovial fibroblasts (RASFs), suppression of pro-inflammatory cytokines or chemokines, anti-angiogenesis as well as protective effects on bone and cartilage destruction will also be discussed. Current literatures strongly imply HDAC inhibitors as innovative anti-rheumatic drug candidates. However, long-term safety is a major concern. Future investigations should focus on identification of molecular anti-rheumatic mechanisms, development of new classes of HDAC inhibitors with better safety and selectivity profiles, combination of HDAC inhibitors with disease modifying anti-rheumatic drugs (DMARDs) and establishment of topical or intra-articular formulations. PMID- 18393883 TI - Rooteomics: the challenge of discovering plant defense-related proteins in roots. AB - In recent years, a strong emphasis has been given in deciphering the function of genes unraveled by the completion of several genome sequencing projects. In plants, functional genomics has been massively used in order to search for gene products of agronomic relevance. As far as root-pathogen interactions are concerned, several genes are recognized to provide tolerance/resistance against potential invaders. However, very few proteins have been identified by using current proteomic approaches. One of the major drawbacks for the successful analysis of root proteomes is the inherent characteristics of this tissue, which include low volume content and high concentration of interfering substances such as pigments and phenolic compounds. The proteome analysis of plant-pathogen interactions provides important information about the global proteins expressed in roots in response to biotic stresses. Moreover, several pathogenic proteins superimpose the plant proteome and can be identified and used as targets for the control of viruses, bacteria, fungi and nematode pathogens. The present review focuses on advances in different proteomic strategies dedicated to the challenging analysis of plant defense proteins expressed during bacteria-, fungi- and nematode-root interactions. Recent developments, limitations of the current techniques, and technological perspectives for root proteomics aiming at the identification of resistance-related proteins are discussed. PMID- 18393884 TI - Structure-based drug design targeting biosynthesis of isoprenoids: a crystallographic state of the art of the involved enzymes. AB - Biosynthesis of the universal terpenoid precursors, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), from three acetyl CoA moieties through mevalonate was studied extensively in the 1950s. For several decades, the mevalonate paradigm reigned supreme and a mevalonate origin was attributed to a growing number of natural products, in many cases erroneously. Besides this biosynthetic pathway, the existence of a second one leading to IPP and DMAPP through 1-deoxy-D-xylulose 5-phosphate and 2C-methyl-D-erythritol 4-phosphate was discovered more recently in plants and some eubacteria. This pathway is widely distributed in the bacterial kingdom including major human pathogens, such as Mycobacterium tuberculosis or Helicobacter pylori and is also essential in the malaria vector Plasmodium falciparum. During the last few years, the genes, enzymes, intermediates and mechanisms of the biosynthetic route have been elucidated by a combination of methods including comparative genomics, enzymology, advanced NMR technology and crystallography. The present crystallographic review of enzymes involved in isoprenoid biosynthesis will be useful for understanding the various catalytic mechanisms and could potentially help for structure-based drug design. PMID- 18393885 TI - Engineering the protein folding landscape in gram-negative bacteria. AB - Gram-negative bacteria, especially Escherichia coli, are often the preferred hosts for recombinant protein production because of their fast doubling times, ability to grow to high cell density, propensity for high recombinant protein titers and straightforward protein purification techniques. The utility of simple bacteria in such studies continues to improve as a result of an ever-increasing body of knowledge regarding their native protein biogenesis machinery. From translation on the ribosome to interaction with cytosolic accessory factors to transport across the inner membrane into the periplasmic space, cellular proteins interact with many different types of cellular machinery and each interaction can have a profound effect on the protein folding process. This review addresses key aspects of cellular protein folding, solubility and expression in E. coli with particular focus on the elegant biological machinery that orchestrates the transition from nascent polypeptide to folded, functional protein. Specifically highlighted are a variety of different techniques to intentionally alter the folding environment of the cell as a means to understand and engineer intracellular protein folding and stability. PMID- 18393886 TI - The structural analysis of large noncovalent oxygen binding proteins by MALLS and ESI-MS: a review on annelid hexagonal bilayer hemoglobin and crustacean hemocyanin. AB - Understanding the function of macromolecular complexes is related to a precise knowledge of their structure. These large complexes are often fragile high molecular mass noncovalent multimeric proteins. Classical biochemical methods for determination of their native mass and subunit composition were used to resolve their quaternary structure, sometimes leading to different models. Recently, the development of mass spectrometry and multi-angle laser light scattering (MALLS) has enabled absolute determination of native masses and subunit masses. Electrospray ionization mass spectrometry (ESI-MS) was used in denaturing and native conditions to probe subunit composition and noncovalent assemblies masses up to 2.25 MDa. In a complementary way, MALLS provides mass and size estimation in various aqueous solvents. ESI-MS method can also give insights into post translational modifications (glycosylation, disulfide bridges ). By combining native mass and subunit composition data, structural models can be proposed for large edifices such as annelid extracellular hexagonal bilayer hemoglobins (HBL Hb) and crustacean hemocyanins (Hc). Association/dissociation mechanisms, protein protein interactions, structural diversity among species and environmental adaptations can also be addressed with these methods. With their absolute mass determination, the very high precision of spectrometry and the versatile nature of light scattering, ESI-MS and MALLS have provided a wealth of data helping to resolve parts of controversies for HBL-Hb models and opening access to new fields of investigation in structural diversity and molecular adaptation. In this review we will focus on annelid HBL-Hb and on crustacean Hc and on the original contributions of ESI-MS and MALLS in this field. PMID- 18393887 TI - Molecular dynamics simulations of proteins and peptides: from folding to drug design. AB - Computer simulations of proteins, lipids and nucleic acids at equilibrium have become essentially routine. However, the fact remains that complete sampling of conformational space continues to be a bottle-neck in the field. The challenge for the future is to overcome such problems and use computational approaches to understand recognition and spontaneous self-organization in biomolecular systems (folding, aggregation and assembly of complexes), processes that cannot be directly observed experimentally. In this review, examples illustrating the extent to which simulations can be used to understand these phenomena in biomolecular systems will be presented along with examples of methodological developments to increase our physical understanding of the processes. The study cases will cover the problems of peptide-receptor recognition and the use of the information obtained for the design of new non-peptidic ligands; the study of the folding mechanism of small proteins and finally the study of the initial stages of peptide self-aggregation. PMID- 18393888 TI - Diversity in structure and function of tethering complexes: evidence for different mechanisms in vesicular transport regulation. AB - The term 'tethering factor' has been coined for a heterogeneous group of proteins that all are required for protein trafficking prior to vesicle docking and SNARE mediated membrane fusion. Two groups of tethering factors can be distinguished, long coiled-coil proteins and multi-subunit complexes. To date, eight such protein complexes have been identified in yeast, and they are required for different trafficking steps. Homologous complexes are found in all eukaryotic organisms, but conservation seems to be less strict than for other components of the trafficking machinery. In fact, for most proposed multi-subunit tethers their ability to actually bridge two membranes remains to be shown. Here we discuss recent progress in the structural and functional characterization of tethering complexes and present the emerging view that the different complexes are quite diverse in their structure and the molecular mechanisms underlying their function. TRAPP and the exocyst are the structurally best characterized tethering complexes. Their comparison fails to reveal any similarity on a struc nottural level. Furthermore, the interactions with regulatory Rab GTPases vary, with TRAPP acting as a nucleotide exchange factor and the exocyst being an effector. Considering these differences among the tethering complexes as well as between their yeast and mammalian orthologs which is apparent from recent studies, we suggest that tethering complexes do not mediate a strictly conserved process in vesicular transport but are diverse regulators acting after vesicle budding and prior to membrane fusion. PMID- 18393889 TI - Novel biomaterials in medicinal chemistry and drug discovery. PMID- 18393891 TI - Drug delivery from therapeutic self-assembled monolayers (T-SAMs) on 316L stainless steel. AB - Delivery of therapeutic agents from self-assembled monolayers (SAMs) on 316L stainless steel (SS) has been demonstrated as a viable method to deliver drugs for localized coronary artery stent application. SAMs are highly-ordered, nano sized molecular coatings, adding 1-10 nm thickness to a surface. Hydroxyl terminated alkanethiol SAMs of 11-mercapto-1-undecanol (-OH SAM) were formed on 316L SS with 48 hr immersion in ethanolic solutions. Attachment of ibuprofen (a model drug) to the functional SAMs was carried out in toluene for 5 hrs at 60 degrees C using Novozume-435 as a biocatalyst. SAM formation and subsequent attachment of ibuprofen was characterized collectively using X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and contact angle (CA) measure-ments. The quantitative in vitro release of ibuprofen into a "physiological" buffer solution was characterized using reverse phase HPLC. Drug release kinetics showed that 14.1 microg of ibuprofen eluted over a period of 35 days with 2.7microg being eluted in the first day and the remaining being eluted over a period of 35 days. The drug release kinetics showed an increase in ibuprofen elution that occurred during first 14 days (2.7microg in 1 day to 9.5 microg in 14 days), following which there was a decrease in the rate of elution. Thus, functional SAMs on 316L SS could be used as tethers for drug attachment and could serve as a drug delivery mechanism from stainless steel implants such as coronary artery stents. PMID- 18393890 TI - Surface chemistry influences implant biocompatibility. AB - Implantable medical devices are increasingly important in the practice of modern medicine. Unfortunately, almost all medical devices suffer to a different extent from adverse reactions, including inflammation, fibrosis, thrombosis and infection. To improve the safety and function of many types of medical implants, a major need exists for development of materials that evoked desired tissue responses. Because implant-associated protein adsorption and conformational changes thereafter have been shown to promote immune reactions, rigorous research efforts have been emphasized on the engineering of surface property (physical and chemical characteristics) to reduce protein adsorption and cell interactions and subsequently improve implant biocompatibility. This brief review is aimed to summarize the past efforts and our recent knowledge about the influence of surface functionality on protein:cell:biomaterial interactions. It is our belief that detailed understandings of bioactivity of surface functionality provide an easy, economic, and specific approach for the future rational design of implantable medical devices with desired tissue reactivity and, hopefully, wound healing capability. PMID- 18393892 TI - A cellular perspective to bioceramic scaffolds for bone tissue engineering: the state of the art. AB - A vast number of manufacturing techniques have been employed in the last five years to manufacture three dimensional (3D) calcium phosphate (CaP) scaffolds, with the intention to replicate the architecture of native bone as well as to repair and restore bone function. Design features such as architectural control and sintering temperature and their impact on scaffold performance is presented in this review. In vitro cell responses to bioceramic scaffolds and their in vivo performances have been enhanced. Current frontiers of active research on HA scaffolds have included the relationship between fluid flow and mechanotransduction as well as cell signaling pathways that induce endothelial cell recruitment and angiogenesis. Additionally, current research has focused on a better understanding of cell signaling and its environmental cues. The availability of non-invasive and non-destructive quantitative imaging modalities has also become critical in aiding the characterization of scaffolds and predicting scaffold performance. It is thus anticipated that further knowledge gained from this research will allow the overall advancement of scaffolds that can be clinically used to restore large bone defects. PMID- 18393894 TI - Synthetic sustained gene delivery systems. AB - Gene therapy today is hampered by the need of a safe and efficient gene delivery system that can provide a sustained therapeutic effect without cytotoxicity or unwanted immune responses. Bolus gene delivery in solution results in the loss of delivered factors via lymphatic system and may cause undesired effects by the escape of bioactive molecules to distant sites. Controlled gene delivery systems, acting as localized depot of genes, provide an extended sustained release of genes, giving prolonged maintenance of the therapeutic level of encoded proteins. They also limit the DNA degradation in the nuclease rich extra-cellular environment. While attempts have been made to adapt existing controlled drug delivery technologies, more novel approaches are being investigated for controlled gene delivery. DNA encapsulated in nano/micro spheres of polymers have been administered systemically/orally to be taken up by the targeted tissues and provide sustained release once internalized. Alternatively, DNA entrapped in hydrogels or scaffolds have been injected/implanted in tissues/cavities as platforms for gene delivery. The present review examines these different modalities for sustained delivery of viral and non-viral gene-delivery vectors. Design parameters and release mechanisms of different systems made with synthetic or natural polymers are presented along with their prospective applications and opportunities for continuous development. PMID- 18393893 TI - Vascularization of engineered tissues: approaches to promote angio-genesis in biomaterials. AB - Although there have been extensive research efforts to create functional tissues and organs, most successes in tissue engineering have been limited to avascular or thin tissues. The major hurdle in development of more complex tissues lies in the formation of vascular networks capable of delivering oxygen and nutrients throughout the engineered constructs. Sufficient neovascularization in scaffold materials can be achieved through coordinated application of angiogenic factors with proper cell types in biomaterials. This review present the current research developments in the design of biomaterials and their biochemical and biochemical modifications to produce vascularized tissue constructs. PMID- 18393895 TI - Enhancing polysaccharide-mediated delivery of nucleic acids through functionalization with secondary and tertiary amines. AB - Chitosan is a polysaccharide that has generated significant interest as a non viral gene delivery vehicle due to its cationic and biocompatible characteristics. However, transfection efficiency of chitosan is significantly lower compared to other cationic gene delivery agents, e.g. polyethyleneimine (PEI), dendrimers or cationic lipids. This is primarily attributed to its minimal solubility and low buffering capacity at physiological pH leading to poor endosomal escape of the gene carrier and inefficient cytoplasmic decoupling of the complexed nucleic acid. Here we have developed an imidazole acetic acid (IAA) modified chitosan to introduce secondary and tertiary amines to the polymer in order to improve its endosomal buffering and solubility. The modified polymer was characterized by ninhydrin and (1)H NMR assays for degree of modification, while buffering and solubility were analyzed by acid titration. Nanocomplex formation, studied at various polymer-nucleic acid ratios, showed an increase in particle zeta potential for chitosan-IAA, as well as an increase in the effective diameter. Up to 100-fold increase in transfection efficiency of pDNA was seen for chitosan-IAA as compared to native chitosan, nearly matching that of PEI. In addition, transfection of siRNA by the modified polymers showed efficient gene knockdown equivalent to commercially available siPORT Amines. Collectively, these results demonstrate the potential of the imidazole-grafted chitosan as a biocompatible and effective delivery vehicle for both pDNA and siRNA. PMID- 18393896 TI - Improved biomaterials for tissue engineering applications: surface modification of polymers. AB - Tissue engineering approaches that combine biomaterial-based scaffolds with protein delivery systems have provided a potential strategy for improved regeneration of damaged tissue. The success of polymeric scaffolds is determined by the response it elicits from the surrounding biological environment. This response is governed, to a large extent, by the surface properties of the scaffold. Surfaces of polymeric scaffolds have a significant effect on protein and cell attachment. Multiple approaches have been developed to provide micrometer to nanometer scale alterations in surface architecture of scaffolds to enable improved protein and cell interactions. Chemical modification of polymeric scaffold surfaces is one of the upcoming approaches that enables enhanced biocompatibility while providing a delivery vehicle for proteins. Similarly, physical adsorption, radiation mediated modifications, grafting, and protein modifications are other methods that have been employed successfully for alterations of surface properties of polymeric scaffolds. The goal of this review is to provide an overview of the role of surface properties /chemistry in tissue engineering and briefly discuss some of the methods of surface modification that can provide improved cell and protein interactions. It is hoped that these improved polymeric scaffolds will lead to accelerated and functional tissue regeneration. PMID- 18393897 TI - Biologically active chitosan systems for tissue engineering and regenerative medicine. AB - Biodegradable polymeric scaffolds are widely used as a temporary extracellular matrix in tissue engineering and regenerative medicine. By physical adsorption of biomolecules on scaffold surface, physical entrapment of biomolecules in polymer microspheres or hydrogels, and chemical immobilization of oligopeptides or proteins on biomaterials, biologically active biomaterials and scaffolds can be derived. These bioactive systems show great potential in tissue engineering in rendering bioactivity and/or specificity to scaffolds. This review highlights some of the biologically active chitosan systems for tissue engineering application and the associated strategies to develop such bioactive chitosan systems. PMID- 18393898 TI - Recent advances in the biochemistry and treatment of cysticercosis. PMID- 18393899 TI - Cysticercosis in the pig. AB - Taenia solium cysticercosis is still an important parasitosis in rural pigs in many developing countries, Mexico among them. The main causes for the persistence of this condition are lack of hygiene in the rural communities, lack of education of the animal owners, lack of control in the trade of pigs and their meat and lack of conscientious meat inspection. The pig production systems in the marginated areas of Mexico are briefly mentioned and it is stressed that among the important reasons for the persistence of the reproductive cycle of Taenia solium is the fact that appropriate toilet facilities in village dwellings are not mandatory. The diagnostic methods of cysticercosis in the living pigs and in their meat are discussed and the degenerative stages of the larvae as well as methods to test their viability are explained. The treatment of infected pigs and their meat is discussed. Recommendations for control programmes are given. PMID- 18393900 TI - Morphology and biochemistry of the pork tapeworm, Taenia solium. AB - This chapter describes the life cycle, general morphology and ultrastructure of the larval and adult stages of Taenia solium, a parasitic flatworm of humans found in underdeveloped countries. Experimental results describing the role of proteins and glycoproteins in the host-parasite relationship, as well as the various strategies the larval stage has developed to evade the host immune responses are analyzed. Characteristics of the tapeworm attachment site in the hamster intestine and the host inflammatory reaction are reviewed. The general morphology and ultrastructure of the experimental tapeworm is described, with emphasis on muscle fiber distribution, the abundance of cytoplasmic glycogen and its association with gap junctions, the development of testis, structure of mature spermatids and vas efferens. Recent descriptions of T. solium actin, myosin and calreticulin components, metabolic steroid pathways, apoptosis and glucose uptake of tapeworms in the hamster model are reviewed. PMID- 18393901 TI - The complex role of pro- and anti-inflammatory cytokines in cysticercosis: immunological lessons from experimental and natural hosts. AB - Parasitic helminthes have developed complex mechanisms to evade or modulate hosts responses. Studies on cysticercosis are solid, but scarce. The most studied immunological models of cysticercosis are Taenia crassiceps infecting mice and T. solium infecting pigs. These parasites, despite being widely exposed to the host, are able to modulate the host immune system. Taenia metacestodes, much like other helminthes parasites, have developed complicated strategies in order to infect and successfully colonize their hosts. We focus here on the accumulated evidence from experimental models that have been helpful in analyzing and characterizing the host immune response to cysticercosis. Moreover, the mouse model has been used to design rationale vaccine strategies, some of them with promising results. We also discuss recent advances in understanding immune-regulation of cysticercosis. The parasite is able to manipulate the host immune system into supporting its survival by keeping a low inflammatory profile by causing the production of some cysticerci-released products that have immunomodulatory activities, as well as promoting the raise of alternatively activated macrophages. Finally, we delineate, according to recent literature, the likely pathway involved in protection and susceptibility against cysticercosis. As more aspects of the role of different immune and parasite-derived molecules are elucidated, better therapeutic targets may be identified to help treat cysticercosis. PMID- 18393902 TI - Taenia solium: antioxidant metabolism enzymes as targets for cestocidal drugs and vaccines. AB - This review focuses in the role that antioxidant enzymes play in protection and other important physiological functions such as signal transduction, cell differentiation, growth and apoptosis. Parasites use these enzymes to evade ROS produced by the host immune response and for development inside the host. In the cestoda Taenia solium, three antioxidant enzymes have been studied: a cystosolic Cu,Zn superoxide dismutase that is a target of cestocidal drugs (bencimidazoles); a 2-Cys peroxiredoxin that is a regulatory enzyme of H(2)O(2), molecule essential for several physiological functions; and two isoforms of glutathione transferases that are immunological targets, since they protect immunized mice against cysticercosis. Moreover, all these enzymes are present in all stages of the parasite. These findings suggest that antioxidant enzymes have an important role in T. solium physiology and infection, therefore they might represent the Achilles' heel of the parasite. PMID- 18393903 TI - The neuroimmunoendocrine network in the complex host-parasite relationship during murine cysticercosis. AB - We review here the role that sex steroids play in experimental intraperitoneal Taenia crassiceps cysticercosis of male and female BalbC/AnN mice. Briefly, estrogens favour and androgens hinder the reproduction of cysticerci by at least two main mechanisms: 1) through estradiol tilting the TH2/TH1 immune system balance towards parasite-permissive TH2 response,which is IL-6 dependent mediating P450-aromatase over expression, shunting testosterone towards estradiol and thus creating a positive feed-back loop which progressively favours TH2 response, blocking in turns TH1 and furthers parasite growth; and 2) estrogens and androgens acting directly upon the cysticercus reproductive system, favoring or hindering, respectively, its asexual reproduction. Later infection, when parasite loads are for milliars, male mice become estrogenized, deandrogenized and diminish their copulative, aggressive and social behaviors in association with P450-aromatase testis overexpression. Changes in c-fos expression in different areas of the infected mice brain point to the additional connection of the central nervous system with the infection driven events, which senses and perhaps reacts to infection with behavioral changes. This complex immune-neuro endocrine network management of parasite loads in murine cysticercosis, and its physiological and behavioral consequences upon the host, may be operative in other infections of mammals. Such complexity may also help to explain the often conflicting results, observed between infections with respect to the role of the host sex, and hints to other avenues of research and strategies for their treatment and control. PMID- 18393904 TI - Are hormones relevant for the search and design of anti-parasitic drugs? AB - Over the last years the biology of many parasites that infect humans and domestic animals has been intensively studied. Considerable efforts were addressed to obtain information on the parasite-host immune relationship. However, the knowledge of the endocrine physiology of parasites and the consequences of the local hormone production on the host tissues needs further investigation. We review here literature and our own studies on endocrine parasite capacities with special emphasis on cysticercosis. Besides the biological interest, these investigations may contribute to identify in the future alternative treatments for the disease. PMID- 18393905 TI - Vaccines against cysticercosis. AB - Taenia solium cysticercosis is a major parasitic disease that seriously and frequently affects human health and economy in undeveloped countries. Since pigs are an indispensable intermediate host, it is conceivable to curb transmission by reducing pig cysticercosis through their effective vaccination. This article reviews current knowledge on the development vaccines against porcine cysticercosis. It highlights the development of several versions of S3Pvac aimed to increase effectiveness, reduce costs and increase feasibility by novel delivery systems and alternative routes of administration. PMID- 18393906 TI - Medical treatment for neurocysticercosis: drugs, indications and perspectives. AB - Neurocysticercosis is one of the most frequent parasitic diseases affecting the central nervous system. The introduction of anticysticidal therapy in the early 80's and the concomitant improvement of the radiological techniques have lead to apparently significant progress in patient prognosis. However, due to the specificity of the disease, a great debate has been generated on the real usefulness of cysticidal drugs. This article revises and discusses the pharmacological aspects of cysticidal treatment and summarizes current indications for the different types of the disease. PMID- 18393907 TI - Angiogenesis: a promising treatment option for peripheral arterial disease. PMID- 18393908 TI - Has the time come for a new definition of microalbuminuria? PMID- 18393909 TI - Mitochondrial MMP activation, dysfunction and arrhythmogenesis in hyperhomocysteinemia. AB - Chronic volume/pressure overload-induced heart failure augments oxidative stress and activates matrix metalloproteinase which causes endocardial endothelial myocyte (EM) uncoupling eventually leading to decline in myocardial systolic and diastolic function. The elevated levels of homocysteine (Hcy), hyperhomocysteinemia (HHcy), are associated with decline in cardiac performance. Hcy impairs the EM functions associated with the induction of ventricular hypertrophy leading to cardiac stiffness and diastolic heart failure. Hcy-induced neurological defects are mediated by the NMDA-R (N-methyl-D-aspartate (NMDA) receptor) activation. NMDA-R is expressed in the heart. However, the role of NMDA R on cardiac function during HHcy is still in its infancy. The blockade of NMDA-R attenuates NMDA-agonist-induced increase in the heart rate. Hcy increases intracellular calcium and activates calpain and calpain-associated mitochondrial (mt) abnormalities have been identified in HHcy. Mitochondrial permeabilization and uncoupling in the pathological setting is fueled by redox stress and calcium mishandling. Recently the role of cyclophilin D, a component of the mitochondrial membrane permeability transition pore, has been identified in cardiac-ischemia. Mechanisms underlying the potentiation between NMDA-R activation and mitochondrial defects leading to cardiac dysfunction during HHcy remain to be elucidated. This review addresses the mitochondrial mechanism by which Hcy contributes to the decline in mechano-electrical function and arrhythmogenesis via agonizing NMDA-R. The putative role of mitochondrial MMP activation, protease stress and mitochondrial permeability transition in cardiac conduction during HHcy is discussed. The review suggests that Hcy increases calcium overload and oxidative stress in the mitochondria and amplifies the activation of mtMMP, causing the opening of mitochondrial permeability transition pore leading to mechano-electrical dysfunction. PMID- 18393910 TI - Atherogenesis in renal patients: a model of vascular disease? AB - Chronic kidney disease (CKD), and particularly kidney failure, is associated with accelerated atherosclerosis and approximately a 20-fold increased risk of cardiovascular death. The majority of these patients die from complications directly attributed to atherosclerosis and their life expectancy is decreased. Established risk factors are involved in the pathogenesis of this phenomenon. Age, gender, smoking, hypertension, dyslipidaemia and diabetes mellitus are among the established risk factors. Inflammation, qualitative lipid disorders (e.g. small dense low density lipoprotein), vascular calcification and oxidative stress represent emerging risk factors. The precise mechanism of atherosclerosis in patients with kidney failure is not yet known. CKD might represent a clinical model of atherogenesis. Thus, the evidence obtained from investigating "renal" atherogenesis could be of interest in improving our understanding of this disease process in the non-renal population. We review the relationship between "renal" and non-renal atherosclerosis focusing on pathogenesis, risk factors and clinical events and how they interact with treatment options. Overall, the "later" stages of CKD may eventually be considered as a coronary heart disease equivalent condition. PMID- 18393912 TI - Cardiac protection by volatile anaesthetics: a review. AB - Ischaemic preconditioning, a response to brief sublethal episodes of ischaemia leading to a pronounced protection against subsequent lethal ischaemia, is mimicked by some pharmacological agents. Halogenated anaesthetics alone exhibit cardioprotective properties at therapeutic doses, independent of their anaesthetic and haemodynamic effect, leading to the concept of anaesthetic preconditioning. Only recently has research turned to clinical application of preconditioning protocols, and anaesthetic preconditioning has indeed been demonstrated in randomised clinical trials conducted in patients undergoing cardiac surgery - mostly coronary artery bypass graft. Most of these trials demonstrate cardiac protection by assessing postprocedural release of cardiac troponin or early postoperative cardiac function. Few studies focus on clinical outcomes, and none demonstrates an advantage in terms of mortality or cardiac morbidity. A recent meta-analysis, pooling data regarding the use of desflurane and sevoflurane, found significant reductions of in-hospital mortality, myocardial infarction rate, intensive care unit and hospital stay, time on mechanical ventilation and incidence of long term cardiac events. In conclusion, the use of desflurane and sevoflurane appears to yield a better outcome, in terms of mortality and cardiac morbidity, in patients undergoing cardiac surgery. A definitive demonstration of this concept represents a difficult task because of the low mortality rate in modern cardiac surgery and because of the number of interfering factors. Whether these cardioprotective properties also exist in non coronary surgery settings is still controversial owing to the scarce available data. PMID- 18393913 TI - Hormone replacement therapy and stroke. AB - Stroke is the third most common cause of death in women and a major cause of disability. Stroke occurs in older age in women compared with men. High premenopausal estrogen concentrations in women are thought to be protective against stroke and cardiovascular disease. Estrogens are essential for normal reproductive function and they exert complex and diverse non reproductive actions on multiple tissues such as neuroprotective effects, vasodilatation, improved vascular reactivity, antithrombotic effects and lipid lowering effects. After menopause estrogen concentrations are depleted and in the past estrogen replacement therapy was considered as a potential protective agent against both cardiovascular disease and stroke. Although the use of hormone therapy was originally associated with a reduction in the risk of heart disease by about 50% in observational studies, the results regarding stroke have been less clear. In order to investigate the effect of hormone therapy on stroke risk, randomized controlled trials of cardio-and/or cerebrovascular-disease prevention in women with established heart disease have been designed. The Heart Estrogen-Progestin Replacement Study included stroke as secondary outcome. This study did not show any differences in myocardial infarction (MI) or coronary death (HR 0.99; 95%CI 0.80-1.22) and in stroke rate. In another study, the Women Estrogen Stroke Trial, 17 beta estradiol 1 mg/placebo was administered to women with previous ischemic stroke or transient ischaemic attack (TIA) having a mean age 71. No differences in stroke rate (RR 1.1; 95% CI 0.8-1.4) and in mortality rate (RR 1.2; 95% CI 0.8 1.8) were found, while a trend showing an increased rate of fatal strokes (RR 2.9; 95% CI 0.9-9.0) and for more severe non-fatal strokes (% patients with final National Institutes of Health Stroke Scale (NIHSS) 0-1: 19 % vs. 33%; p = 0.12) was observed. The Women's Health Initiative, a primary prevention study, where conjugated equine estrogen (CEE) plus medroxyprogesterone acetate/placebo was utilized, was stopped because of an excess in breast cancer and increased stroke rates (RR 1.4; 95% CI 1.1-1.8). Recently, a meta-analysis including 39,769 women participating in 28 trials has been published. Twelve studies were of secondary prevention and the overall stroke rate was 2%. In the hormone replacement therapy (HRT) arm there was a 29% increased rate of ischemic stroke (Number Needed to Harm, NNH:147). Furthermore, a 56% increased rate of death or dependency after stroke and a tendency of more fatal stroke were observed. Additionally, a higher stroke risk was reported in the first year of treatment. CONCLUSIONS: There seems to be no indication for hormone replacement therapy in the prevention of stroke in women. Further studies are needed to discover why estrogens have different effects on the heart and brain. Conventional risk-factors which could increase the risk of estrogen therapy need to be identified and as well as more restrictive inclusion and exclusion criteria such as coagulation parameters and intimal thickness should be adopted before new randomized trials are started. PMID- 18393914 TI - Statins for the prevention of first or recurrent stroke. AB - This review considers the evidence showing that statins can prevent first or recurrent stroke or improve its outcome in subjects at moderate or high risk for cardiovascular disease (CVD). Data are reviewed according to trial design (observational or prospective) and baseline CVD risk. Two (ASCOT, CARDS) out of five primary CVD prevention statin trials showed a considerable reduction in stroke rates. In two (MIRACL and PROVE IT) out of five acute coronary syndrome trials, the prevention of first stroke was significant. Most secondary prevention trials (4S, CARE, LIPID, HPS, GREACE and TNT) showed a beneficial effect of statins in stroke prevention. Finally, SPARCL, the only secondary stroke prevention trial in subjects without overt coronary heart disease (CHD), showed a significant reduction in total and ischaemic (fatal and nonfatal) stroke rate, although a small but significant increase in nonfatal haemorrhagic stroke was noted. There was also a significant reduction in CHD-related events. The possible mechanisms responsible for statin-associated stroke prevention are discussed. The evidence suggests the need to consider early and long-term statin treatment (with substantial low-density lipoprotein cholesterol reduction) in all patients at high risk of any type of major vascular event, without discriminating CHD from stroke. Thus, statins may be beneficial to both the heart and the brain. PMID- 18393915 TI - Fondaparinux for the prevention or treatment of venous thromboembolism related to lower limb trauma: evidence today. AB - Patients with lower limb and pelvic trauma, or undergoing major orthopaedic surgery represent one of the highest risk groups for the development of venous thromboembolism (VTE). A significant number of pharmacological and mechanical agents have been used for the prophylaxis and treatment of VTE. Fondaparinux is a relative new pharmacological agent that selectively binds to antithrombin, and represents a new class of synthetic selective inhibitors of activated factor X. Eleven percent of the fondaparinux-related English language literature, between 2001 and 2007, refers to orthopaedic trauma, and was the sample assessed for this critical analysis review. The clinical studies evaluating the safety, efficacy, and financial implications associated with lower limb orthopaedic trauma show that fondaparinux has comparable results with the well-established use of enoxaparin. However, the scientific community has raised several issues regarding mostly fondaparinux's safety, timing of its 1(st) dose, bleeding side effects, duration of administration and lack of a reliable reversing agent. Further trials are necessary focusing on the safety and efficacy of this drug mostly in relation to clinical relevant outcomes and to different fields of trauma surgery (pelvis, long bone fractures and polytrauma patients). PMID- 18393916 TI - Current status of carotid stenting. AB - Stroke is the third most common cause of death worldwide following ischemic heart disease and cancer and the number one condition associated with permanent disability. In Western countries, the yearly incidence of stroke is approximately 0.2% of the population and the number of stroke-related death is expected to double over the next 30 years. Studies from the Netherlands and Scotland showed that stroke accounted for 3% to 5% of their total health care resources. For 2006, the estimated direct and indirect cost of stroke in the United States was approximately 60 billion. Significant stenosis of the internal carotid artery is responsible for 10% to 20% of all strokes or transient ischemic attacks. Large scale randomized clinical trials have established the superiority of carotid endarterectomy (CEA) over medical management in patients with high-grade stenosis of the internal carotid artery, particularly among symptomatic patients. Although percutaneous carotid revascularization has been performed since the early eighties, the enthusiasm has long been tempered by the fear of cerebral embolism. Following the introduction of emboli protection devices (EPD) around the year 2000, the number of procedures performed worldwide has grown exponentially. We review the available studies on carotid stenting and discuss appropriate use of this procedure. Emphasis is also placed on optimal long-term pharmacotherapy in patients with carotid stenosis. PMID- 18393918 TI - Possible roles of reg family proteins in pancreatic islet cell growth. AB - Reg proteins constitute a conserved family in human and rodents; their production in the pancreas (including the islets of Langerhans) is induced upon beta-cell damage. While some members of the family (Reg1 and islet neogenesis-associated protein, i.e. INGAP) have been implicated in beta-cell replication and/or neogenesis, including from in vivo studies using transgenic and knockout mice; the roles of the other five members have yet to be characterized. Among them, Reg2 was recently proposed to serve as an autoantigen on beta-cells that elicits T-cell attack in type 1 diabetes mellitus. Elucidation of their actions and identification of their molecular targets should provide insight into the biology of these proteins and lead to the design and development of novel strategies aimed at promoting the survival and function of the pancreatic islets. As the current terminology used for mammalian Reg genes/proteins is very confusing, we also proposed a uniformed classification in human and rodents through sequence alignments. PMID- 18393917 TI - Pathogenesis and treatment of secondary hyperparathyroidism in dialysis patients: the role of paricalcitol. AB - Hemodialysis (HD) patients are commonly affected by secondary hyperparathyroidism (SHPT), in which 3 well-known factors are usually involved: hypocalcemia, hyperphosphatemia and calcitriol deficiency. Classically, high parathyroid hormone (PTH) levels cause bone-associated diseases, such as osteitis fibrosa and renal osteodystrophy, but more recently it has been demonstrated the link between SHPT and a systemic toxicity, with a major role in determining cardio-vascular disease, including arterial calcification, endocrine disturbances, compromised immune system, neurobehavioral changes, and altered erythropoiesis. Treatment with calcitriol (CT), the active form of vitamin D, reduces parathyroid hormone (PTH) levels, but may result in elevations in serum calcium (Ca) and phosphorus (P), increasing the risk of cardio-vascular calcification in the HD population. Several new vitamin D analogs have been developed and investigated with the rationale to treat SHPT with a reduced risk of hypercalcemia and hyperphosphatemia in HD patients. Paricalcitol (1,25-dihydroxy-19-nor-vitamin D(2), 19-Nor-D(2)) is a vitamin D analog that suppresses PTH secretion with minimal increases on serum calcium and phosphate levels. It was demonstrated that paricalcitol prevents vascular calcification in experimental models of renal failure, compared with calcitriol. Furthermore, 19-Nor-D(2) is the first analog approved for use in HD patients and is available for i.v. and oral administration, commonly 3 times weekly after HD. The purpose of the present review is to analyze the pathogenesis and treatment of SHPT in HD patients, and the role of paricalcitol in the prevention of arterial calcification. PMID- 18393919 TI - The use of mycophenolate mofetil for the treatment of systemic sclerosis. AB - Mycophenolate mofetil (MMF) is an inosine monophosphate dehydrogenase inhibitor, that inhibits the de novo pathway of guanosine nucleotide synthesis, the proliferative responses of T and B lymphocytes as well as antibody production by B-lymphocytes. It is indicated for the prophylaxis of organ rejection after allogeneic cardiac, hepatic and renal transplants . It has recently also been used with good success in patients with lupus nephritis . Based on these actions, MMF appears to be a novel agent for the treatment of systemic sclerosis, especially during early disease where an inflammatory infiltrate preceeds the development of fibrosis. Disease modification early on during the inflammatory stage of systemic sclerosis may lead to an overall decrease in fibrotic complications both in relation to cutaneous and internal organ involvement. PMID- 18393920 TI - Host-pathogen interactions in latent Mycobacterium tuberculosis infection: identification of new targets for tuberculosis intervention. AB - Mycobacterium tuberculosis (M. tuberculosis) is one of the worlds' most successful and sophisticated pathogens. It is estimated that over 2 billion people today harbour latent M. tuberculosis infection without any clinical symptoms. Since most new cases of active tuberculosis (TB) arise from this (growing) number of latently infected individuals, urgent measures to control TB reactivation are required, including more effective drugs and new TB vaccines. The currently widely used BCG vaccines, as well as most new generation TB vaccines that are being developed are designed as prophylactic or as BCG-booster vaccines. Unfortunately, many of these vaccines are unlikely to be effective in individuals already latently infected with M. tuberculosis. Here we argue that detailed analysis of M. tuberculosis genes that are switched on predominantly during the latent stage of infection may lead to the identification of new M. tuberculosis targets for drug and vaccine development. First, we will describe essential host-pathogen interactions in TB with particular emphasis on TB latency and persistent infection. Subsequently, we will focus on a novel group of late stage specific genes, encoded by the M. tuberculosis dormancy (dosR) regulon, and summarize recent studies describing human T-cell recognition of these dormancy antigens in relation to (latent) M. tuberculosis infection. We will discuss the possible relevance of these new classes of antigens for new TB intervention strategies. PMID- 18393921 TI - The HIV-1 Rev binding family of proteins: the dog proteins as a study model. AB - Various proteins that are required for the building of new complete human immunodeficiency type 1 virions (HIV-1) are coded by unspliced or partly spliced virus-derived mRNAs. HIV-1 has developed special strategies for moving these mRNAs to the cytoplasm to be translated. In the nucleus of the infected cell the virus-derived protein Regulator of expression of viral proteins (Rev) can bind both the viral intron-containing mRNAs and the cellular co-factor HIV-1 Rev binding protein (HRB) and this complex may be shuttled through the nuclear pores. HRB genes have been relatively well conserved during evolution, from Drosophila to humans. However, as a consequence of reading-frame shifts due to nt insertions/deletions, the protein products generated may differ considerably from the prototypal HRB protein, which comprises one Arf-GAP zinc finger domain, several Phenylalanine-Glycine (FG) motifs and four Asparagine-Proline Phenylalanine (NPF) motifs. This variability is best exemplified by four HRB proteins of the dog, which are discussed here in more detail. The hypothesis is advanced that atypical HRB proteins may not be able to bind Rev and possibly have other, still undetermined, functions. Since the cellular co-factor HRB is essential for viral replication and spread but is not required for cell viability and main bodily functions, it might be an attractive candidate for anti-HIV-1 drug targeting. PMID- 18393922 TI - Distinct functions of retinoic acid receptor beta isoforms: implications for targeted therapy. AB - Vitamin A is essential for development and differentiation of multiple tissues. Its derivatives, the retinoids, are potent drugs used to treat and prevent a variety of diseases. Retinoid effects are mediated by retinoic acid receptors (RARs) and retinoid X receptors (RXRs). There are three known RARs (alpha, beta, and gamma), and multiple isoforms of each receptor exist. Many of the therapeutic effects of retinoids including cancer chemoprevention and treatment of dermatologic disorders are mediated through RARbeta. In humans, five isoforms of this gene have been described. Specific isoforms of RARbeta exert distinct and sometimes opposing functions by altering patterns of target gene induction. Functional isoforms that activate distinct cassettes of target genes with differing biologic consequences include RARbeta1' and RARbeta2. Dominant negative isoforms of this gene that inhibit target gene activation include RARbeta4 and RARbeta5. RARbeta1 is poorly understood although this may function as an oncogene in certain cancers. Chromatin modifying drugs have been shown to trigger isoform specific changes in the RARbeta gene. This review focuses on the structure and function of RARbeta isoforms as well as recent work in the epigenetic targeting of specific RARbeta isoforms. Discerning isoform-specific functions will be critical for exploiting the full potential of retinoid-based therapy including rational approaches to combining retinoids with chromatin modifying drugs. PMID- 18393923 TI - G protein coupled receptors as drug targets: the role of beta-arrestins. AB - G protein coupled receptors (GPCRs) are extremely important drug targets and the beta-arrestin intracellular scaffolding and adaptor proteins regulate major aspects of their pharmacology. beta-arrestin binding to activated, GPCR kinase (GRK)-phosphorylated receptors has the capacity to terminate G protein coupling, internalize the receptors into clathrin-coated vesicles and establish a secondary signaling complex independent of G protein signaling. These events appear to be differentially regulated by GRK phosphorylation, ubiquitination and potentially beta-arrestin oligomerization, which are likely to be highly receptor and cell type dependent. The role of beta-arrestins in switching from G-protein dependent to independent signaling places them in a pivotal position to dictate the downstream effects of ligand binding. Consequently, we must appreciate the functioning of these molecules as we strive to discover and optimize new GPCR drug therapies for endocrine, metabolic and immune disorders. PMID- 18393924 TI - Monoclonal antibodies: new therapeutic agents for autoimmune hemolytic anemia? AB - Conventional treatment of autoimmune hemolytic anemia (AIHA) comprises corticosteroids and splenectomy and/or other immunosuppressive drugs for refractory/relapsed patients. Monoclonal antibodies (MoAbs) rituximab and alemtuzumab have gained widespread acceptance in the management of B-cell malignancies. More recently, they have been used to treat a number of autoantibody-mediated diseases, such as both idiopathic and secondary AIHA, with encouraging results. Herein we report an overview of the medical literature on the use of MoAbs to treat AIHA. The therapeutic mechanism of action of rituximab in the treatment of autoimmune diseases such as rheumatoid arthritis and lupus is currently a subject of considerable investigation. We have proposed that cell associated IgG immune complexes, generated by the binding of rituximab to CD20 on B cells, may serve as decoys that attract FcgammaR-expressing effector cells and downregulate effector cell pathogenic action, thus reducing inflammation and tissue destruction in these diseases. We briefly review evidence that suggests that this immune complex decoy hypothesis plays a role in the therapeutic action of rituximab in AIHA, and we propose new measurements that should allow for a more complete evaluation of the importance of this mechanism in AIHA. PMID- 18393933 TI - Eye muscle surgery for nystagmus. RE: Outcome study of two standard and graduated augmented modified kestenbaum surgery protocols for abnormal head postures in infantile nystagmus. Binocul Vis Strabismus Q 2007; 22:235-41. PMID- 18393934 TI - Reading new electronic version of BVSQ. PMID- 18393925 TI - Epithelial-mesenchymal transition as a therapeutic target for prevention of ocular tissue fibrosis. AB - Fibrotic diseases are characterized by the appearance of myofibroblasts, the key cell type involved in the fibrogenic reaction, and by excess accumulation of extracellular matrix with resultant tissue contraction and impaired function. Myofiborblasts are generated by fibroblast-myofibrobalst conversion, and in certain tissues through epithelial-mesenchymal transition (EMT), a process through which an epithelial cell changes its phenotype to become more like a mesenchymal cell. Although inflammatory/fibrogenic growth factors/cytokines produced by injured tissues orchestrate the process of EMT, transforming growth factor beta (TGFbeta) is believed to play a central role in the process. Unlike fibrotic lesions in kidney or other tissues where myofibroblasts are generated from both fibroblasts and epithelial cells, fibrotic lesions in the eye crystalline lens are derived only from lens epithelial cells without contamination of fibroblast-derived myofibroblasts. Thus, this tissue is suitable to investigate detailed mechanisms of EMT and subsequent tissue fibrosis. EMT in retinal pigment epithelium is involved in the development of another ocular fibrotic disease, proliferative vitreoretinopathy, a fibrosis in the retina. EMT related signal transduction cascades, i. e., TGFbeta/Smad, are a target to prevent or treat unfavorable ocular tissue fibrosis, e. g., fibrotic diseases in the crystalline lens or retina, as well as possibly in other organs. PMID- 18393935 TI - Due to TV? Critiques and moral changes in our society; e-publishing; binocular vision vertical fusion; unique esotropia; papilledema and cystinosis. PMID- 18393936 TI - Response pattern asymmetry of binocular vision vertical fusion amplitudes in a normal population. AB - PURPOSE: To compare the maximum vertical fusion amplitudes in vertical directions (response pattern by respective higher eye), to investigate for possible asymmetry. METHODS: Vertical fusion amplitude (VFA) was measured in one hundred normal adults with normal single binocular vision using a computer based device that produced a gradually increasing, but optically distant test distance, Vertical fusion amplitudes in the vertical left- over-right (L/R, elevation of the left eye above the right eye) pattern direction were compared to the vertical fusion amplitudes in the vertical right-over-left direction pattern(R/L). RESULTS: The mean VFA was 4.08 +/-1.01 prism diopters, with vertical fusion amplitudes of 4.40 +/-1.44 prism diopters in the L/R direction and 3.75 +/-1.19 prism diopters in the R/L direction. There was no association between ocular dominance and the direction of greater VFA. Asymmetry in the vertical fusion response was found, with 77/100 subjects showing greater than a 10% difference between the means of the L/R and R/L measurements. The group with asymmetry had 17% greater mean VFA than the symmetric group. CONCLUSION: Normal adults frequently have asymmetric directional vertical fusion amplitudes. The asymmetry appears to be due to the contribution of the direction with the greater VFA, rather than the result of both alternatives. These results are intriguing and bear further investigation. Possible explanations my be asymmetry of orbital anatomy or functional asymmetry of either muscular or neuronal origin. They do not appear to be due to the relative ocular dominance. Inherent VFA asymmetry should therefore be considered in all forms of vergence testing in the vertical direction. PMID- 18393937 TI - An unusual case of adult progressive esotropia caused by high myopia. AB - BACKGROUND: In progressive esotropia associated with high myopia and axial elongation, eso-hypodeviation of the eyeball occurs due to ocular dislocation and often progresses to complete fixed esotropia in the terminal stage. We report a rare case of this condition in whom manual pushing of the eyeball temporarily moved the ocular dislocation back into the muscle cone. A normal eye position and ocular movement were obtained in subsequent strabismus surgery. To our knowledge, there has been no previous report of such a case. It is uncertain if medial rectus muscle recession should be performed simultaneously with combination of the muscle bellies of the superior and lateral rectus muscles in surgery for progressive esotropia caused by high myopia. We discuss this issue in the context of the current case. CASE REPORT: The patient was a 60 year old woman with a chief complaint of severe eso-hypotropia of the left eye, for which requested treatment. Ophthalmologic findings include refractive indices of -5.15 D right eye and left eye -22.0 D respectively. The left eye position was severely eso hypotropic and ocular movement was limited in all directions. However, the left eye became capable of abduction when the medial side of the eye was pushed manually by rubbing during attempted levoversion. During levoversion while the patient was pushing the eyeball, the dislocation was reduced on Computerized Tomography imaging. In surgery, left medial rectus muscle recession and combination of the muscle bellies of the left superior rectus muscle and the left lateral rectus muscle were performed. After surgery, the eye position was markedly corrected and the abduction limitation was improved. CONCLUSION: We encountered a case of progressive esotropia caused by high myopia in which ocular dislocation could be temporarily reversed. In this disease, pushing of the eyeball (push test) can be used to determine whether dislocation ban be temporarily reversed. If this is possible, determination of the degree of abduction may be useful for selection of an appropriate surgical procedure. PMID- 18393938 TI - A case of intracranial hypertension and papilledema associated with nephropathic cystinosis and ocular involvement. AB - An 11 year old boy with nephropathic cystinosis developed moderate to severe bilateral optic disc edema two months after he received a deceased donor renal allograft. The bilateral optic disc edema was found to be a result of intracranial hypertension diagnosed by lumbar puncture. No etiology was found. He was treated with acetazolamide and his optic dis edema resolved over a period of eight months and did not recur after acetazolamide was discontinued. The mechanism of intracranial hypertension in patients with nephropathic cystinosis is not well understood, but may involve obstruction of cerebrospinal fluid outflow due to deposition of cystine crystals in arachnoid villi. PMID- 18393939 TI - Regulation of hypoxia-inducible factor-1alpha by NF-kappaB. AB - HIF (hypoxia-inducible factor) is the main transcription factor activated by low oxygen tensions. HIF-1alpha (and other alpha subunits) is tightly controlled mostly at the protein level, through the concerted action of a class of enzymes called PHDs (prolyl hydroxylases) 1, 2 and 3. Most of the knowledge of HIF derives from studies following hypoxic stress; however, HIF-1alpha stabilization is also found in non-hypoxic conditions through an unknown mechanism. In the present study, we demonstrate that NF-kappaB (nuclear factor kappaB) is a direct modulator of HIF-1alpha expression. The HIF-1alpha promoter is responsive to selective NF-kappaB subunits. siRNA (small interfering RNA) studies for individual NF-kappaB members revealed differential effects on HIF-1alpha mRNA levels, indicating that NF-kappaB can regulate basal HIF-1alpha expression. Finally, when endogenous NF-kappaB is induced by TNFalpha (tumour necrosis factor alpha) treatment, HIF-1alpha levels also change in an NF-kappaB-dependent manner. In conclusion, we find that NF-kappaB can regulate basal TNFalpha and, in certain circumstances, the hypoxia-induced HIF-1alpha. PMID- 18393940 TI - Biochemical function of typical and variant Arabidopsis thaliana U-box E3 ubiquitin-protein ligases. AB - The variance of the U-box domain in 64 Arabidopsis thaliana (thale cress) E3s (ubiquitin-protein ligases) was used to examine the interactions between E3s and E2s (ubiquitin-conjugating enzymes). E2s and E3s are components of the ubiquitin protein degradation pathway. Seven U-box proteins were analysed for their ability to ubiquitinate proteins in vitro in co-operation with different E2s. All U-box domains exhibited ubiquitination activity and interacted productively with UBC4/5 type E2s. Three and four of the U-box domains mediated ubiquitin addition in the presence of UBC13 and UBC7 E2s respectively, but no productive interaction was observed with the UBC15 E2 tested. The activity of AtPUB54 [Arabidopsis thaliana (thale cress) plant U-box 54 protein] was dependent on Trp(266) in the E2-binding cleft, and the E2 selectivity was changed by substitution of this position. The function of the distant U-box protein, AtPUB49, representing a large family of eukaryotic proteins containing a U-box linked to a cyclophilin-like peptidyl prolyl cis-trans isomerase domain, was characterized biochemically. AtPUB49 functioned both as a prolyl isomerase and a chaperone by catalysing cis-trans isomerization of peptidyl-prolyl bonds and dissolving protein aggregates. In conclusion, both typical and atypical Arabidopsis U-box proteins were active E3s. The overlap in the E3/E2 selectivity suggests that in vivo specificity is not determined only by the E3-E2 interactions, but also by other parameters, e.g. co existence or interactions with additional domains. The biochemical functions of AtPUB49 suggest that the protein can be involved in folding or degradation of protein substrates. Similar functions can also be retained within a protein complex with separate chaperone and U-box proteins. PMID- 18393941 TI - Relationship between birthweight and arterial elasticity in childhood. AB - There is a considerable debate about the potential influence of 'fetal programming' on cardiovascular diseases in adulthood. In the present prospective epidemiological cohort study, the relationship between birthweight and arterial elasticity in 472 children between 5 and 8 years of age was assessed. LAEI (large artery elasticity index), SAEI (small artery elasticity index) and BP (blood pressure) were assessed using the HDI/PulseWave CR-2000 CardioVascular Profiling System. Blood concentrations of glucose, total cholesterol and its fractions [LDL (low-density lipoprotein)-cholesterol and HDL (high-density lipoprotein) cholesterol] and triacylglycerols (triglycerides) were determined by automated enzymatic methods. Insulin was assessed by a chemiluminescent method, insulin resistance by HOMA (homoeostasis model assessment) and CRP (C-reactive protein) by immunonephelometry. Two linear regression models were applied to investigate the relationship between the outcomes, LAEI and SAEI, and the following variables: birthweight, gestational age, glucose, LDL-cholesterol, HDL cholesterol, triacylglycerols, insulin, CRP, HOMA, age, gender, waist circumference, per capita income, SBP (systolic BP) and DBP (diastolic BP). LAEI was positively associated with birthweight (P=0.036), waist circumference (P<0.001) and age (P<0.001), and negatively associated with CRP (P=0.024) and SBP (P<0.001). SAEI was positively associated with birthweight (P=0.04), waist circumference (P=0.001) and age (P<0.001), and negatively associated with DBP (P<0.001). Arterial elasticity was decreased in apparently healthy children who had lower birthweights, indicating an earlier atherogenetic susceptibility to cardiovascular diseases in adolescence and adult life. Possible explanations for the results include changes in angiogenesis during critical phases of intrauterine life caused by periods of fetal growth inhibition and local haemodynamic anomalies as a way of adaptation to abnormal pressure and flow. PMID- 18393942 TI - Analysis of mutant origin recognition complex with reduced ATPase activity in vivo and in vitro. AB - In eukaryotes, ORC (origin recognition complex), a six-protein complex, is the most likely initiator of chromosomal DNA replication. ORC belongs to the AAA(+) (ATPases associated with a variety of cellular activities) family of proteins and has intrinsic ATPase activity derived from Orc1p, one of its subunits. To reveal the role of this ATPase activity in Saccharomyces cerevisiae (baker's yeast) ORC, we mutated the Orc1p sensor 1 and sensor 2 regions, which are important for ATPase activity in AAA(+) proteins. Plasmid-shuffling analysis revealed that Asn(600), Arg(694) and Arg(704) are essential for the function of Orc1p. In yeast cells, overexpression of Orc1R694Ep inhibited growth, caused inefficient loading of MCM (mini-chromosome maintenance complex of proteins) and slowed the progression of S phase. In vitro, purified ORC-1R [ORC with Orc1R694Ep (Orc1p Arg(694)-->Glu mutant)] has decreased ATPase activity in the presence or absence of origin DNA. However, other activities (ATP binding and origin DNA binding) were indistinguishable from those of wild-type ORC. The present study showed that Arg(694) of the Orc1p subunit is important for the ATPase activity of ORC and suggests that this ATPase activity is required for efficient MCM loading on to origin DNA and for progression of S phase. PMID- 18393944 TI - Cold storage of pooled, buffy-coat-derived, leucoreduced platelets in plasma. AB - BACKGROUND: This study was designed to determine which in vitro assays would be most useful for studying the effects of cold storage on platelet concentrates and to establish an in vivo model for platelet recovery and survival. STUDY DESIGN AND METHODS: Paired, plasma-suspended, leucoreduced, buffy-coat-derived platelet concentrates were stored either at 22 or 4 degrees C. Prior to storage and after 18 h, 5 days and 7 days, samples were taken and various assays were performed. On day 6, in vivo studies were carried out using a model system. Galactosylation of the platelets, prior to cold storage, was also tested. RESULTS: Hypotonic shock response, collagen-induced aggregation, RANTES and P-selectin binding site measurements demonstrated differences between platelets stored at 22 and 4 degrees C. The glycocalicin assay was able to demonstrate microvesicle formation at 4 degrees C. The in vivo model showed that there was at least a 50% decrease in recovery and survival when the platelets were stored in the cold. Galactosylation did not improve these results. CONCLUSIONS: Several assays, both in vitro and in vivo, were able to detect differences in platelet-storage characteristics and in vivo recovery and survival in a model system. Galactosylation did not correct these cold-induced changes. PMID- 18393943 TI - Specific effects of KChIP3/calsenilin/DREAM, but not KChIPs 1, 2 and 4, on calcium signalling and regulated secretion in PC12 cells. AB - The KChIPs (K+ channel-interacting proteins) are members of the NCS (neuronal calcium sensor) protein family of Ca2+-binding proteins. It is unclear to what extent the KChIPs have distinct functions although they all interact with Kv4 K+ channels. KChIP3 has also been shown to repress transcription of specific genes via binding to DRE (downstream regulatory element) motifs and all KChIPs may share this function. In the present study, we have compared the function of isoforms of the four KChIPs. KChIPs 1-4 were found to stimulate the traffic of Kv4.2 channels to the plasma membrane. KChIP3 expression in PC12 cells resulted in an increase in exocytosis evoked by activation of purinergic receptors. In contrast, KChIPs 1, 2 and 4, although expressed to the same extent, had no effect on secretion. In addition, KChIP3 but not KChIPs 1, 2 and 4 modified the ATP induced Ca2+ signal resulting in a delay in recovery after the peak Ca2+ elevation and also specifically resulted in down-regulation of the Na+/Ca2+ exchanger NCX3, which could explain the effects on the Ca2+ signal and secretion. Regulation of NCX3 by KChIP3 has been shown to occur via its DREAM (DRE antagonist modulator) function [Gomez-Villafuertes, Torres, Barrio, Savignac, Gabellini, Rizzato, Pintado, Gutierrez-Adan, Mellstrom, Carafoli and Naranjo (2005) J. Neurosci. 25, 10822-10830] suggesting that this activity might depend on the cellular context of expression of the various KChIPs. These results reveal a new role for KChIP3 in the regulation of Ca2+-regulated secretion and also suggest that the functions of each of the KChIPs may be more specialized than previously appreciated. PMID- 18393945 TI - Screening platelet concentrates for bacterial contamination: low numbers of bacteria and slow growth in contaminated units mandate an alternative approach to product safety. AB - BACKGROUND AND OBJECTIVES: We introduced 100% screening of platelets for bacterial contamination in 2005 to reduce the risk of clinical sepsis from platelet transfusion. We test all outdating units again at expiry to assess the sensitivity of the initial test. MATERIALS AND METHODS: We test all platelet concentrates prior to release for clinical use using a large volume automated culture technique on the day after manufacture. All units that expire unused are retested. Platelets still in stock on day 4 of storage may have a repeat culture performed, and are returned to stock with two extra days of shelf life. RESULTS: Of 43,230 platelet units screened, 35 (0.08%) were positive; of 8282 expired unused, 18 (0.22%) were positive; and of 3310 day-4 retests, four (0.12%) were positive. Overall sensitivity of the initial screening test was 29.2% (95% confidence interval 19.4 to 39.1%). Thirteen of the 35 positive screening tests would have been expected to grow in both aerobic and anaerobic bottles; eight grew in aerobic culture only and five grew in anaerobic culture only, indicating that the likely number of bacteria in the contaminated platelet units at the time of sampling was less than 60 colony-forming unit per platelet unit. CONCLUSIONS: Screening platelet concentrates for bacterial contamination using the most sensitive method available has a sensitivity of less than 40% because of the low numbers of bacteria in the initial contamination. Effective resolution of this problem will require a pathogen-inactivation technique. PMID- 18393946 TI - Hepatitis G virus: a suitable marker of in vivo efficacy for pathogen inactivation. AB - Being an orphan virus despite a large number of investigations, hepatitis G virus is a blood-borne agent for which screening is not required in blood donations. The in vivo efficacy of pathogen inactivation methods could be assessed by the absence of hepatitis G virus markers after transfusion of pathogen-inactivated blood products, in recipients susceptible to infection before the transfusion. PMID- 18393947 TI - Tubes for pretransfusion testing should be collected by blood bank staff and hand labelled until the implementation of new technology for improved sample labelling. Results of a prospective study. AB - BACKGROUND AND OBJECTIVES: The greatest risk in transfusion medicine is actually human error, resulting in the use of the incorrect blood component. The aim of our study was to identify and evaluate the risk factors involved in the collection and labelling of pretransfusion blood samples. MATERIAL AND METHODS: We prospectively evaluated 6446 samples submitted to the blood bank for pretransfusion testing. Inappropriate samples were classified as 'mislabelled' or 'miscollected'. After 4 months of study, an educational approach was taken. RESULTS: The frequency of inappropriately labelled samples was 6.45%. Such samples were associated with the use of addressograph labels (vs. hand-written labels) [23.4% vs. 1.4%, P < 0.0001], collection by clinical staff (vs. blood bank staff) [8.8% vs. 2.1%, P = 0.001] and emergency situations (vs. routine sampling) [10.1% vs. 6.1%, P = 0.005]. Following educational intervention, the percentage of inappropriately labelled samples decreased from 7.3% (pre educational) to 5.8% (post-educational), P = 0.005. CONCLUSION: Ongoing monitoring and analysis of labelling and collection should be mandatory in order to improve the safety of transfusion. PMID- 18393948 TI - A novel platform for biologically active recombinant human interleukin-13 production. AB - Interleukin-13 (IL-13) is a pleiotropic regulatory cytokine with the potential for treating several human diseases, including type-1 diabetes. Thus far, conventional expression systems for recombinant IL-13 production have proven difficult and are limited by efficiency. In this study, transgenic plants were used as a novel expression platform for the production of human IL-13 (hIL-13). DNA constructs containing hIL-13 cDNA were introduced into tobacco plants. Transcriptional expression of the hIL-13 gene in transgenic plants was confirmed by reverse transcriptase-polymerase chain reaction and Northern blotting. Western blot analysis showed that the hIL-13 protein was efficiently accumulated in transgenic plants and present in multiple molecular forms, with an expression level as high as 0.15% of total soluble protein in leaves. The multiple forms of plant-derived recombinant hIL-13 (rhIL-13) are a result of differential N-linked glycosylation, as revealed by enzymatic and chemical deglycosylation, but not of disulphide-linked oligomerization. In vitro trypsin digestion indicated that plant rhIL-13 was more resistant than unglycosylated control rhIL-13 to proteolysis. The stability of plant rhIL-13 to digestion was further supported with simulated gastric and intestinal fluid digestion. In vitro bioassays using a factor-dependent human erythroleukaemic cell line (TF-1 cells) showed that plant rhIL-13 retained the biological functions of the authentic hIL-13 protein. These results demonstrate that transgenic plants are superior to conventional cell based expression systems for the production of rhIL-13. Moreover, transgenic plants synthesizing high levels of rhIL-13 may prove to be an attractive delivery system for direct oral administration of IL-13 in the treatment of clinical diseases such as type-1 diabetes. PMID- 18393949 TI - A tattoo-pigmented node masquerading as the sentinel node in a case of breast cancer. PMID- 18393950 TI - Is the Perigord black truffle threatened by an invasive species? We dreaded it and it has happened! PMID- 18393951 TI - Mixed-linkage (1-->3,1-->4)-beta-D-glucan is a major hemicellulose of Equisetum (horsetail) cell walls. AB - Mixed-linkage (1-->3,1-->4)-beta-d-glucan (MLG) is a hemicellulose reputedly confined to certain Poales. Here, the taxonomic distribution of MLG, and xyloglucan, especially in early-diverging pteridophytes, has been re investigated. Polysaccharides were digested with lichenase and xyloglucan endoglucanase (XEG), which specifically hydrolyse MLG and xyloglucan, respectively. The oligosaccharides produced were analysed by thin-layer chromatography (TLC), high-pressure liquid chromatography (HPLC) and alkaline peeling. Lichenase yielded oligo-beta-glucans from all Equisetum species tested (Equisetum arvense, Equisetum fluviatile, Equisetum scirpoides, Equisetum sylvaticum and Equisetum xtrachyodon). The major product was the tetrasaccharide beta-glucosyl-(1-->4)-beta-glucosyl-(1-->4)-beta-glucosyl-(1-->3)-glucose (G4G4G3G), which was converted to cellotriose by alkali, confirming its structure. Minor products included G3G, G4G3G and a nonasaccharide. By contrast, poalean MLGs yielded G4G3G > G4G4G3G > nonasaccharide > dodecasaccharide. No other pteridophytes tested contained MLG, including Psilotum and eusporangiate ferns. No MLG was found in lycopodiophytes, bryophytes, Chara or Nitella. XEG digestion showed that Equisetum xyloglucan has unusual repeat units. Equisetum, an exceedingly isolated genus whose closest living relatives diverged > 380 million years ago, has evolved MLG independently of the Poales. Equisetum and poalean MLGs share basic structural motifs but also exhibit clear-cut differences. Equisetum MLG is firmly wall-bound, and may tether neighbouring microfibrils. It is also suggested that MLG acts as a template for silica deposition, characteristic of grasses and horsetails. PMID- 18393952 TI - Empty microcatheter technique for the deployment of a self-expanding stent to treat refractory middle cerebral artery occlusion in the setting of severe proximal tortuosity. AB - We report a case of an acute middle cerebral artery (M2 segment) occlusion that was refractory to intravenous tissue plasminogen activator (IV t-PA), thrombectomy with the Merci device, intra-arterial infusion of urokinase and eptifibatide, and balloon angioplasty. The artery was so tortuous that over-the wire stent placement failed and a salvage technique was required to place the self-expanding stent. PMID- 18393953 TI - Granulomatosis with CNS involvement: a neuroimaging clinicopathologic correlation. AB - BACKGROUND AND PURPOSE: To present a rare case of a rheumatologic disorder with a central nervous system manifestation. METHODS: A 48-year-old female with history of nasal and auricular pain and inflammation developed bilateral intermittent scleritis that was refractory to several immunosuppressive medications. She later presented with headaches and right upper extremity parasthesias. Brain magnetic resonance imaging (MRI) was obtained and she was referred to a neurosurgeon. RESULTS: Neurological examination did not reveal any focal deficits. Laboratory analysis demonstrated an elevated erythrocyte sedimentation rate (ESR). Brain MRI with and without gadolinium contrast revealed an extra-axial, enhancing, left frontoparietal convexity lesion. The patient was taken to the operating room for biopsy and resection of the mass. Final pathology demonstrated dural granulomatosis with vasculitis, most consistent with intracranial Wegener's granulomatosis. The patient was discharged to follow-up with her rheumatologist for further medical management. CONCLUSIONS: Enhancing, intracranial dural lesions are associated with a variety of disease entities. Wegener's granulomatosis is one such entity and should be considered in the correct context. PMID- 18393954 TI - The relationship between the level of consciousness and sedimentation level in CT scan of chronic subdural hematoma. PMID- 18393955 TI - Studies on the value of diffusion-weighted MR imaging in the early prediction of periventricular leukomalacia. AB - OBJECTIVE: Periventricular leukomalacia (PVL) is the most common cerebral injury in premature infants. While cranial ultrasonography and conventional magnetic resonance imaging (MRI) offer little for its early diagnosis, still they are much favored at present. Based on the pathologic mechanism of ischemic cerebral injury in PVL, we attempt to elucidate the effects of diffusion-weighted MR imaging (DWI) for the early diagnosis of PVL. METHODS: Twelve cases of PVL in premature infants confirmed by MRI examinations were included in the current study. All cases underwent DWI in addition to conventional MR imaging (T1-weighted [T1W] and T2-weighted [T2W]) 7 days, 2 weeks, and 4 approximately 8 weeks after birth, respectively. RESULTS: Initial DWI examination (carried out at an average of 4.5 days after birth) revealed symmetrical and diffuse high signal in bilateral periventricular areas in all cases, while conventional MRI demonstrated normal signal. DWI examination of 2 weeks later showed irregular inhomogeneous signals in cerebral white matter, while conventional MRI revealed hyperintensity on T1WI and slight hypointensity on T2WI in corresponding areas. Four weeks later, DWI revealed various intensities of cystic low signals beside lateral ventricles, while conventional MRI showed hypointensity on T1WI and hyperintensity on T2WI in corresponding lesion, ie, change in cystic PVL. Four months later, conventional MRI evidenced gradual shrink and disappearance of cystic cavity, reduction in cerebral white matter, and ventricle enlargement. CONCLUSIONS: Symmetrical, diffuse hyperintensity in both lateral periventricular white matter on DWI was the earliest sign of PVL in premature infants. Imaging abnormalities provided by DWI had a close correlation with the results obtained by conventional MRI performed at the advanced stage of PVL, which suggests that DWI may be an important sequence for the early evaluation of cerebral white matter injury as well as for the prediction of PVL occurrence in premature infants. PMID- 18393956 TI - Consistency of the CFAE phenomena using custom software for automated detection of complex fractionated atrial electrograms (CFAEs) in the left atrium during atrial fibrillation. AB - INTRODUCTION: Complex fractionated atrial electrograms (CFAEs) have been described as a potential target for ablation of atrial fibrillation (AF). The purpose of this study is to assess the consistency of the CFAE phenomena using custom software for automated detection of CFAEs in the left atrium during AF. METHODS AND RESULTS: This prospective study included 10 patients referred for catheter ablation of symptomatic drug-refractory AF. Ten consecutive points at a single location (cluster) were acquired as electroanatomical points every 3 seconds. Atrial signals were automatically classified as CFAEs by the software algorithm. The number of intervals between 50 ms and 110 ms and in the voltage range 0.05-0.15 mV during the 2.5-second recordings was determined and referred to as the interval confidence level (ICL). A total of 2,226 points were acquired during mapping of AF. A dominant group of ICL using one of two different configurations of ICL fractionation was identified. A dominant group was defined as the ICL categorization occurring with greatest frequency in a given cluster of points. The results show the consistency ranged from 73%+/- 21 for the three group configuration (ICL < or = 4; 4 < ICL < or = 7; ICL > 7) to 84%+/- 16 for the two-group configuration (ICL < or = 5; ICL > 5). CONCLUSION: This novel software offers an objective method for CFAE analysis during atrial fibrillation. CFAE consistency ranged from 73% to 84% with wide standard deviation. Automated detection of CFAEs may remove the pitfalls associated with subjective visual detection, thus removing one variable in comparative studies of using CFAEs as AF ablation targets. PMID- 18393957 TI - The psychometric properties of the Hospital Anxiety and Depressions Scale adapted for use with people with intellectual disabilities. AB - BACKGROUND: There is increasing recognition of depression in people with intellectual disabilities (ID). There is a need to develop well-standardised self report measures for both clinical and research purposes. This paper presents some psychometric properties of the Hospital Anxiety and Depression Scale (HADS) adapted for use with people with ID. The anhedonic emphasis in the depression scale of the HADS may be particularly useful for people with ID who present with high rates of physical co-morbidity. METHOD: A total of 197 people with ID completed an adapted HADS; 32 participants also completed the Zung Depression Scale and 42 also completed the Glasgow Depression Scale. RESULTS: The obtained factor structure is similar to the original form of the scale used with people without ID. However, the underlying question wording of the HADS, where most depression items are phrased positively and most anxiety items are phrased negatively, makes clear interpretation of the factor structure difficult. The HADS has an adequate internal reliability and correlates well with other self report measures of depression. CONCLUSIONS: The HADS may need further adaptation; however, the measurement of anhedonia is a useful addition to the self-report measures of depression available for people with ID. PMID- 18393958 TI - Virulence blockers as alternatives to antibiotics: type III secretion inhibitors against Gram-negative bacteria. AB - In recent years mounting problems related to antibiotic-resistant bacteria have resulted in the prediction that we are entering the preantibiotic era. A way of preventing such a development would be to introduce novel antibacterial medicines with modes of action distinct from conventional antibiotics. Recent studies of bacterial virulence factors and toxins have resulted in increased understanding of the way in which pathogenic bacteria manipulate host cellular processes. This knowledge may now be used to develop novel antibacterial medicines that disarm pathogenic bacteria. The type III secretion system (T3SS) is known to be a potent virulence mechanism shared by a broad spectrum of pathogenic Gram-negative bacteria that interact with human, animal and plant hosts by injecting effector proteins into the cytosol of host cells. Diseases, such as bubonic plague, shigellosis, salmonellosis, typhoid fever, pulmonary infections, sexually transmitted chlamydia and diarrhoea largely depend on the bacterial proteins injected by the T3SS machinery. Recently a number of T3SS inhibitors have been identified using screening-based approaches. One class of inhibitors, the salicylidene acylhydrazides, has been subjected to chemical optimization and evaluation in several in vitro and ex vivo assays in multiple bacterial species including Yersinia spp., Chlamydia spp., Salmonella spp. and Pseudotuberculosis aeruginosa. Reports published up to date indicate that T3SS inhibitors have the potential to be developed into novel antibacterial therapeutics. PMID- 18393960 TI - Nonattendance in a dermatology clinic--a large sample analysis. AB - BACKGROUND: Previous studies have described factors determining non-attendance at dermatology appointments in small sample sizes. OBJECTIVE: To perform an analysis of factors associated with non-attendance in a dermatology clinic in a larger sample. METHODS: Factors determining non-attendance were examined in 52 604 consecutive first-time visits to a dermatology clinic over a period of 44 months. RESULTS: Non-attendance proportion was 27.6%. Among children, non-attendance was associated with waiting for an appointment < 7 days [odds ratio (OR), 1.44], Bedouin sector (OR, 1.30), rural Jewish sector (OR, 0.45) and the treating physician. Among adults, non-attendance was associated with female gender (OR, 1.08), age < 55 years (OR, 1.65), waiting time for an appointment < 7 days (OR, 1.44), timing of the appointment between 1 and 4 pm (OR, 1.13), Bedouin sector (OR, 1.63), rural Jewish sector (OR, 0.46) and the treating physician. CONCLUSION: Non-attendance is common among Bedouins, adult female patients and young adults and is more likely as waiting times become longer. Strategies to reduce non-attendance are needed. PMID- 18393959 TI - Risk of hypertension amongst Swedish male snuff users: a prospective study. AB - BACKGROUND: The scientific evidence on whether long-term use of snuff is associated with high blood pressure is limited, inconsistent and based only on cross-sectional data. OBJECTIVE: We aimed at studying the risk of hypertension in relation to long-term use of snuff based on longitudinal data. DESIGN: Repeated health check-ups were offered to all employees in the Swedish construction industry between 1978 and 1993. Blood pressure was measured at the health check up and information on tobacco use and other risk factors was collected through questionnaires. SETTING: In total, 120 930 never smoking men with information on blood pressure and snuff use at baseline were included. The association of high blood pressure and snuff use at baseline was estimated by logistic regression. Further, 42 055 men were identified as normotensive at baseline and had at least one subsequent health check-up. Through repeated blood pressure measurements and linkage to the Swedish National Inpatient Register, information on hypertension was obtained. Relative risk estimates were derived from Cox proportional hazards regression model. RESULTS: Almost 30% of all men had used snuff. The adjusted odds ratio of high blood pressure amongst snuff users at baseline was 1.23 (95% CI 1.15-1.33) compared to never snuff users. The relative risk of high blood pressure during follow-up was 1.39 (95% CI 1.08-1.79) amongst snuff users and 1.36 (95% CI 1.07-1.72) for hypertension as recorded in the Inpatient Register. CONCLUSION: Use of Swedish moist snuff appears to be associated with a moderately increased risk of hypertension. PMID- 18393961 TI - Association between oral lichenoid reactions and amalgam restorations. AB - BACKGROUND: The aim of this study was to perform a clinical assessment of the association between oral lichenoid reactions (OLR) and amalgam restorations and to determine the salivary concentrations of interleukin-6 (IL-6) and IL-8 before and after replacement of the amalgam restorations. METHODS: The study included 20 patients with OLR and 20 healthy volunteers, who were examined between 2001 and 2005 at the Oral Medicine Unit of the Medical Faculty University of Rijeka. All patients were skin patch tested by an experienced physician. Saliva samples were collected, prepared and analysed for IL-6 and IL-8 concentrations using enzyme linked immunosorbent assay. RESULTS: Sixteen out of 20 patch-tested patients showed a sensitization to inorganic mercury or amalgam. Total replacement of all amalgam fillings was carried out on 20 patients with fillings based on composite resin, gold, porcelain or a combination of these. Sixteen out of 20 patients showed complete healing of OLR; three patients had marked improvement, whereas one patient showed no improvement. Levels of IL-6 detected before replacement were significantly higher than IL-6 levels following the replacement (P = 0.003). The IL-8 levels measured before replacement procedure were significantly higher than the IL-8 levels after replacement of the fillings (P < 0.001). CONCLUSIONS: On the basis of clinical observations, restorative therapy resulted in tissue healing. Following the replacement of amalgam fillings with fillings based on other restorative materials, levels of both IL-6 and IL-8 shifted towards normal, as measured in healthy subjects. PMID- 18393962 TI - Dardia Lipo Milk improves skin dryness: a corneometric assessment. PMID- 18393963 TI - The polymorphous light eruption-severity assessment score does not reliably predict the results of phototesting. AB - BACKGROUND: Polymorphous light eruption (PLE) is a very common photodermatosis in which patient history is highly specific. Phototesting is used to confirm the diagnosis and to determine the action spectrum and the severity of this disease. In daily practice and in research studies, it would be convenient to assess disease severity by patient history only. OBJECTIVES: This study aims to assess PLE disease severity via patient history and compares this with severity assessment via phototesting. PATIENTS AND METHODS: Sixty-one patients with PLE were asked 10 standard questions and all were phototested. The answers to the standard questions were coded with linear scores ranging from 0 to 10. The score of each question was plotted as independent variable in a multiple linear regression model against the score of the phototest (minimal number of irradiations necessary to elicit a positive skin lesion, with a maximum of 6 irradiations) as dependent variable using an enter approach. Furthermore, the scores of the separate questions were added to form a total score, the PLE severity assessment score (PLE-SAS). The medians of these PLE-SASs were compared with the result scores obtained by phototesting. Phototesting was done with ultraviolet A and ultraviolet B irradiation. RESULTS: Fifty-seven of the 61 patients had a positive test result (93%). Using the multiple linear regression model, the severity assessment by patient history (PLE-SAS) compared with the result of phototesting showed two significant contributing questions (adjusted PLE-SAS) (P < 0.05) but with a regression coefficient of 0.2. A significant difference in median scores with the severity assessment (PLE-SAS and adjusted PLE-SAS) between patients testing positive after 1-3 irradiations compared with those testing positive after 4-6 irradiations was present (P < 0.05). However, the overlap quartile range between both groups was such that the PLE-SAS and the adjusted PLE-SAS have little predictive value in individual patients. CONCLUSIONS: We showed that in PLE, disease severity as determined using the PLE SAS or adjusted PLE-SAS did not reliably predict severity as assessed by phototesting. Two significant contributing questions were not discriminating enough to be used as predicting questions to assess severity. Accurate patient history proved to be a reliable method to diagnose PLE. Phototesting is useful to determine the responsible ultraviolet action spectrum and to exclude differential diagnoses like photosensitive eczema, lupus erythematosus or chronic actinic dermatitis. PLE-SAS cannot replace phototesting for determining the severity of PLE. PMID- 18393964 TI - Bilateral segmental lentiginosis associated with malignant melanomas. PMID- 18393965 TI - Primary care physician beliefs about insulin initiation in patients with type 2 diabetes. AB - BACKGROUND: Insulin is the most effective drug available to achieve glycaemic goals in patients with type 2 diabetes. Yet, there is reluctance among physicians, specifically primary care physicians (PCPs) in the USA, to initiate insulin therapy in these patients. AIMS: To describe PCPs' attitudes about the initiation of insulin in patients with type 2 diabetes and identify areas in which there is a clear lack of consensus. METHODS: Primary care physicians practicing in the USA, seeing 10 or more patients with type 2 diabetes per week, and having > 3 years of clinical practice were surveyed via an internet site. The survey was developed through literature review, qualitative study and expert panel. RESULTS: Primary care physicians (n = 505, mean age = 46 years, 81% male, 62% with > 10 years practice; 52% internal medicine) showed greatest consensus on attitudes regarding risk/benefits of insulin therapy, positive experiences of patients on insulin and patient fears or concerns about initiating insulin. Clear lack of consensus was seen in attitudes about the metabolic effects of insulin, need for insulin therapy, adequacy of self-monitoring blood glucose, time needed for training and potential for hypoglycaemia in elderly patients. CONCLUSIONS: The beliefs of some PCPs are inconsistent with their diabetes treatment goals (HbA1c < or = 7%). Continuing medical education programmes that focus on increasing primary care physician knowledge about the progression of diabetes, the physiological effects of insulin, and tools for successfully initiating insulin in patients with type 2 diabetes are needed. PMID- 18393966 TI - The impact of an assisted diabetes care programme on a private practice. AB - AIMS: Specially trained nurses who follow detailed protocols and algorithms under the supervision of a diabetologist can markedly improve diabetes outcomes in community health centres. We aimed to study the impact of a nurse-assisted diabetes care (NADC) model on diabetes and clinic's financial outcomes in a private practice setting. METHODS: Nurse-assisted diabetes care was provided to the diabetic patients referring to a Monday private diabetes clinic in Shiraz. 107 patients who had received such care were hierarchically matched with 107 diabetic patients receiving usual endocrinologist care in the same clinic during the rest of the week. At the end of 6 months of follow-up, outcomes [glycosylated haemoglobin (HbA1c), serum triglycerides, low-density lipoprotein (LDL) cholesterol, duration of patient's visit and net clinic's income] for patients under NADC were compared with those of usual care patients and also with those derived from the 6 months before receiving NADC. RESULTS: Under NADC, HbA1c levels had a more significant fall (p < 0.03), significantly smaller proportions of patients had triglyceride levels of > 1.69 mmol/l (150 mg/dl) and LDL cholesterol of > 2.58 mmol/l (100 mg/dl) (both p < 0.05), the time for one patient's visit decreased by an average of 9.3 min (p = 0.000) while the clinic's net income increased by 21.25%. CONCLUSION: Nurse-assisted diabetes care, while improving diabetes outcomes significantly, spares time for the physician and allows more patients to be seen per clinic hours. The excess income from extra visits much outweighs the expenditures including the nurses' wages. NADC is profitable for private diabetes clinics or offices. PMID- 18393967 TI - Assessment of osteoporotic fracture risk in community settings: a study of post menopausal women in Australia. AB - The healthcare cost of managing osteoporotic fractures is projected to rise because of the change in population demographics. To reduce the fracture epidemic, strategies are needed to identify those at risk early to allow preventative intervention to be implemented. The aim of this study was to investigate if low-cost community-based assessments, such as calcaneal ultrasound and falls risk assessments, can discriminate a group of elderly women at risk of osteoporotic fracture from those at lower risk. Over the period of 2002-2003, 104 community-dwelling women (mean age 71.3, standard deviation 5.8) were recruited via various modes including advertisements in community newspaper and community centres. These women underwent dual-energy X-ray absorptiometry bone mineral density (BMD) and calcaneal quantitative ultrasound (QUS) measurements; spinal radiography; and performance-based assessment of strength, mobility and balance. The women were classified into a 'high risk' (osteoporotic) group, based on low BMD (T-score of 70% of EMSY amplifications were CCND1 amplified, EMSY may not have any additional effect on survival of ER+ breast cancer. PMID- 18393978 TI - Overexpression of osteopontin in clear cell carcinoma of the ovary: close association with HNF-1beta expression. AB - AIMS: Transcription factor hepatocyte nuclear factor (HNF)-1beta is selectively expressed in clear cell carcinoma (CCC) of the ovary. One of the potential HNF 1beta target genes is osteopontin (OPN). Although elevation of OPN mRNA has been reported in CCC, it remains unclear whether CCC cells overexpress OPN protein. The aim was to investigate the expression of OPN protein and its correlation with HNF-1beta status in CCC. METHODS AND RESULTS: Three CCC and two serous adenocarcinoma (SA) cell lines were evaluated for expression of OPN by reverse transcriptase-polymerase chain reaction and immunocytochemistry. OPN expression, at both the mRNA and protein levels, was higher in the three CCCs than in the two SAs. HNF-1beta expression was detected in the CCCs but not in the SAs. Subsequently, 60 surgical specimens (30 CCCs and 30 SAs) were examined immunohistochemically for expression of OPN and HNF-1beta. All 30 CCCs showed immunopositivity for both OPN and HNF-1beta. The 12 (40%) CCCs with a high OPN score all had a high HNF-1beta score. In contrast, SAs rarely showed immunoreactivity for OPN or HNF-1beta. CONCLUSIONS: OPN expression is elevated in ovarian CCC and is closely associated with HNF-1beta overexpression. HNF-1beta is likely to participate in OPN up-regulation in CCC. PMID- 18393979 TI - Renal cell carcinoma marker reliably discriminates central nervous system haemangioblastoma from brain metastases of renal cell carcinoma. AB - AIMS: The distinction between central nervous system (CNS) metastases of clear cell renal cell carcinoma (RCC) and CNS haemangioblastoma still poses a challenge to the pathologist. Since both entities occur in von Hippel-Lindau disease, this aggravates the issue. The antibody renal cell carcinoma marker (RCC-ma) has been suggested to identify primary RCCs specifically, but its value for diagnosing metastases of RCC is controversial. The aim was to assess two distinct clones of the RCC-ma for their potential to: (i) identify primary RCCs and (ii) differentiate between CNS metastases of clear cell RCC and CNS haemangioblastomas. METHODS AND RESULTS: Using tissue microarrays, 77% (n = 363; PN-15) and 66% (n = 355; 66.4C2) of clear cell RCCs, and 93% (PN-15) and 74% (66.4C2) of papillary RCCs (n = 46) were immunopositive for RCC-ma, whereas none of the investigated chromophobe RCCs (n = 22) or any of the oncocytomas (n = 15) showed immunoreactivity. Importantly, 50.9% of CNS metastases of clear cell RCCs (n = 55) exhibited RCC-ma expression, whereas all CNS haemangioblastomas (71) were negative. CONCLUSIONS: Both RCC-ma clones, despite some variation in their sensitivity to detect clear cell and papillary RCCs, are of value in differentiating subtypes of primary RCC and are excellent markers for discriminating clear cell lesions in the brain. PMID- 18393980 TI - Enhancement of factor VIIa haemostatic efficacy by formulation with PEGylated liposomes. AB - Recombinant activated factor VII (rFVIIa) is an effective treatment of the haemophilia patient with inhibitors and acquired haemophilia. However, on account of its relatively short half-life (HL), achieving therapeutic efficacy with FVIIa requires repeated injections. The development of a long-acting FVIIa product would therefore be beneficial. The formulation of factor VIII with PEGylated liposomes (PEGLip) was previously shown to extend the bleeding-free period in haemophilia patients. We report here an enhancement of haemostatic efficacy by similarly formulating FVIIa with PEGLip. Surface plasmon resonance analysis indicated that FVIIa binds non-covalently but with high affinity and specificity to PEGLip. A one-stage clotting assay demonstrated that formulation of FVIIa with PEGLip does not affect FVIIa activity and stability. A pharmacokinetic study in rats demonstrated that PEGLip formulation of FVIIa extends circulation time and results in higher FVIIa levels several hours after injection. Thromboelastography experiments indicated that PEGLip-FVIIa induces faster clot formation and higher clot stability than standard formulated FVIIa. These results suggest that formulation of FVIIa with PEGLip may lead to a safe and effective long-acting FVIIa that improves the care of haemophilic patients with inhibitors and acquired haemophilia. PMID- 18393981 TI - Establishment of a new mismatch PCR-RFLP technique for detection of G10430A common mutation present in moderate to mild haemophilia B patients belonging to Gujarati community from the western part of India. PMID- 18393982 TI - Painful periarticular calcifications after muscle injury at the hip in a patient with moderate haemophilia A. PMID- 18393983 TI - Health status in Iranian haemophilic patients. PMID- 18393984 TI - Congenital hypofibrinogenemia associated with novel heterozygous fibrinogen Bbeta and gamma chain mutations. PMID- 18393985 TI - Extracellular electron transfer: wires, capacitors, iron lungs, and more. PMID- 18393986 TI - Hard X-ray micro(spectro)scopy: a powerful tool for the geomicrobiologists. AB - During the past few decades, the use of electron microscopy approaches - many developed by Terry Beveridge - to probe the physiology of microorganisms has become a mainstay in fields including microbiology, human health, and geomicrobiology. Recent developments of third-generation synchrotron X-ray sources and X-ray-based microscopy approaches for studying microbial systems have proved their utility as complements to the very powerful approaches regularly employed by electron microscopists. In addition, in recent geomicrobiological studies, researchers have begun to take advantage of the strengths of each technique by using the superior spatial resolution of the electron microscope (relative to the X-ray microscope) and the superior elemental sensitivity of the X-ray microscope (relative to the electron microscope), along with the ability of the X-ray microscope to spatially probe the chemical speciation of elements. The benefits of integrating these two nanoprobes for investigating the same microenvironments within a geomicrobial system are far superior to those of independent studies separately employing each probe. PMID- 18393987 TI - Chemical gene synthesis: strategies, softwares, error corrections, and applications. AB - Chemical synthesis of DNA sequences provides a powerful tool for modifying genes and for studying gene structure, expression and function. Modified genes and consequently protein/enzymes can bridge genomics and proteomics research or facilitate commercial applications of gene and protein technologies. In this review, we will summarize various strategies, designing softwares and error correction methods for chemical gene synthesis, particularly for the synthesis and assembly of long DNA molecules based on polymerase cycling assembly. Also, we will briefly discuss some of the major applications of chemical synthesis of DNA sequences in basic research and applied areas. PMID- 18393988 TI - A new DGGE protocol targeting 2,4-diacetylphloroglucinol biosynthetic gene phlD from phylogenetically contrasted biocontrol pseudomonads for assessment of disease-suppressive soils. AB - In the rhizosphere, biocontrol pseudomonads producing 2,4-diacetylphloroglucinol (Phl) can protect plants from soil-borne pathogens. DGGE of phlD has been proposed to monitor these bacteria, but two distinct protocols were needed for analysis of both the 'Pseudomonas fluorescens' species complex and the strains from rrs restriction group ARDRA-1. Here, a single DGGE protocol performed on 668 bp GC-clamp-containing phlD amplicons was effective with both types of pseudomonads, and 36 reference biocontrol strains from the 'P. fluorescens' complex or group ARDRA-1 gave a total of 11 distinct DGGE bands. phlD amplicons with at least two to seven nucleotidic differences could be discriminated, and the discrimination level was similar to that of phlD restriction analysis with four enzymes. Multiple phlD-DGGE bands were obtained when studying rhizosphere soil containing indigenous phlD+ pseudomonads, and phlD diversity was higher when DGGE was implemented after incubation of tobacco rhizosphere extracts in semi selective medium (MPN approach) in comparison with approaches based on direct analysis of rhizosphere DNA extracts or assessment of phlD+colonies. phlD-DGGE profiles differed for a soil suppressive and a soil conducive to black root rot of tobacco, and each soil yielded new phlD sequences. In conclusion, this DGGE protocol was useful for monitoring indigenous rhizosphere consortia of phlD+ pseudomonads. PMID- 18393989 TI - A new Thraustochytrid, strain Fng1, isolated from the surface mucus of the hermatypic coral Fungia granulosa. AB - Recent evidence suggests that there is a dynamic microbial biota living on the surface and in the mucus layer of many hermatypic coral species that plays an essential role in coral well-being. Most of the studies published to date emphasize the importance of prokaryotic communities associated with the coral mucus in coral health and disease. In this study, we report the presence of a protist (Fng1) in the mucus of the hermatypic coral Fungia granulosa from the Gulf of Eilat. This protist was identified morphologically and molecularly as belonging to the family Thraustochytridae (phylum Stramenopile, order Labyrinthulida), a group of heterotrophs widely distributed in the marine environment. Morphological examination of this strain revealed a nonmotile organism c. 35 mum in diameter, which is able to thrive on carbon-deprived media, and whose growth and morphology are inoculum dependent. Its fatty acid production profile revealed an array of polyunsaturated fatty acids. A similar protist was also isolated from the mucus of the coral Favia sp. In light of these findings, its possible contribution to the coral holobiont is discussed. PMID- 18393991 TI - The rhizobial adhesion protein RapA1 is involved in adsorption of rhizobia to plant roots but not in nodulation. AB - The effect of the rhizobium adhesion protein RapA1 on Rhizobium leguminosarum bv. trifolii adsorption to Trifolium pratense (red clover) roots was investigated. We altered RapA1 production by cloning its encoding gene under the plac promoter into the stable vector pHC60. After introducing this plasmid in R. leguminosarum bv. trifolii, three to four times more RapA1 was produced, and two to five times higher adsorption to red clover roots was obtained, as compared with results for the empty vector. Enhanced adsorption was also observed on soybean and alfalfa roots, not related to R. leguminosarum cross inoculation groups. Although the presence of 1 mM Ca2+ during rhizobial growth enhanced adsorption, it was unrelated to RapA1 level. Similar effects were obtained when the same plasmid was introduced in Rhizobium etli for its adsorption to bean roots. Although root colonization by the RapA1-overproducing strain was also higher, nodulation was not enhanced. In addition, in vitro biofilm formation was similar to the wild type both on polar and on hydrophobic surfaces. These results suggest that RapA1 receptors are present in root but not on inert surfaces, and that the function of this protein is related to rhizosphere colonization. PMID- 18393990 TI - Potential for atmospheric deposition of bacteria to influence bacterioplankton communities. AB - Biogeographic patterns in microbial communities are an exciting but controversial topic in microbial ecology. Advances in theory pertaining to assembly of microbial communities have made strong assumptions about dispersal of bacteria without exploration. For this reason, we investigated rates of atmospheric bacterial deposition and compared the taxonomic composition of bacteria in rain with that of common freshwater bacterial communities. Our findings suggest that it is not appropriate to take for granted that atmospheric deposition of bacteria is a significant vector of immigration to freshwater ecosystems. PMID- 18393992 TI - Community composition of ammonia-oxidizing bacteria and archaea in soils under stands of red alder and Douglas fir in Oregon. AB - This study determined nitrification activity and nitrifier community composition in soils under stands of red alder (Alnus rubra) and Douglas fir (Pseudotsuga menziesii) at two sites in Oregon. The H.J. Andrews Experimental Forest, located in the Cascade Mountains of Oregon, has low net N mineralization and gross nitrification rates. Cascade Head Experimental Forest, in the Coast Range, has higher net N mineralization and nitrification rates and soil pH is lower. Communities of putative bacterial [ammonia-oxidizing bacteria (AOB)] and archaeal [ammonia-oxidizing archaea (AOA)] ammonia oxidizers were examined by targeting the gene amoA, which codes for subunit A of ammonia monooxygenase. Nitrification potential was significantly higher in red alder compared with Douglas-fir soil and greater at Cascade Head than H.J. Andrews. Ammonia-oxidizing bacteria amoA genes were amplified from all soils, but AOA amoA genes could only be amplified at Cascade Head. Gene copy numbers of AOB and AOA amoA were similar at Cascade Head regardless of tree type (2.3-6.0 x 10(6)amoA gene copies g(-1) of soil). DNA sequences of amoA revealed that AOB were members of Nitrosospira clusters 1, 2 and 4. Ammonia-oxidizing bacteria community composition, determined by terminal restriction fragment length polymorphism (T-RFLP) profiles, varied among sites and between tree types. Many of the AOA amoA sequences clustered with environmental clones previously obtained from soil; however, several sequences were more similar to clones previously recovered from marine and estuarine sediments. As with AOB, the AOA community composition differed between red alder and Douglas-fir soils. PMID- 18393994 TI - The small genome of an abundant coastal ocean methylotroph. AB - OM43 is a clade of uncultured beta-proteobacteria that is commonly found in environmental nucleic acid sequences from productive coastal ocean ecosystems, and some freshwater environments, but is rarely detected in ocean gyres. Ecological studies associate OM43 with phytoplankton blooms, and evolutionary relationships indicate that they might be methylotrophs. Here we report on the genome sequence and metabolic properties of the first axenic isolate of the OM43 clade, strain HTCC2181, which was obtained using new procedures for culturing cells in natural seawater. We found that this strain is an obligate methylotroph that cannot oxidize methane but can use the oxidized C1 compounds methanol and formaldehyde as sources of carbon and energy. Its complete genome is 1304 428 bp in length, the smallest yet reported for a free-living cell. The HTCC2181 genome includes genes for xanthorhodopsin and retinal biosynthesis, an auxiliary system for producing transmembrane electrochemical potentials from light. The discovery that HTCC2181 is an extremely simple specialist in C1 metabolism suggests an unanticipated, important role for oxidized C1 compounds as substrates for bacterioplankton productivity in coastal ecosystems. PMID- 18393993 TI - Disentangling the rhizosphere effect on nitrate reducers and denitrifiers: insight into the role of root exudates. AB - To determine to which extent root-derived carbon contributes to the effects of plants on nitrate reducers and denitrifiers, four solutions containing different proportions of sugar, organic acids and amino acids mimicking maize root exudates were added daily to soil microcosms at a concentration of 150 microg C g(-1) of soil. Water-amended soils were used as controls. After 1 month, the size and structure of the nitrate reducer and denitrifier communities were analysed using the narG and napA, and the nirK, nirS and nosZ genes as molecular markers respectively. Addition of artificial root exudates (ARE) did not strongly affect the structure or the density of nitrate reducer and denitrifier communities whereas potential nitrate reductase and denitrification activities were stimulated by the addition of root exudates. An effect of ARE composition was also observed on N(2)O production with an N(2)O:(N(2)O + N(2)) ratio of 0.3 in microcosms amended with ARE containing 80% of sugar and of 1 in microcosms amended with ARE containing 40% of sugar. Our study indicated that ARE stimulated nitrate reduction or denitrification activity with increases in the range of those observed with the whole plant. Furthermore, we demonstrated that the composition of the ARE affected the nature of the end-product of denitrification and could thus have a putative impact on greenhouse gas emissions. PMID- 18393995 TI - Is routine changing of peripheral arterial catheters justified? AB - Although peripheral arterial catheters (pACs) are used extensively, disagreement persists concerning the practice of scheduled replacement to prevent catheter related infections. Despite recommendations and no proof of benefit, pAC replacement continues to be scheduled as a routine practice in many intensive care units (ICUs) worldwide. Our own experience in an oncology ICU, based on a 217-device database, confirms that the risk for pAC-related infections is stable over time, arguing against scheduled replacement. The low rate and stability of the risk of pAC-related infections supports the rationale for conservative management in accordance with expert recommendations. PMID- 18393996 TI - Effect on diagnostic yield of repeated stool testing during outbreaks of Clostridium difficile-associated disease. AB - The effect on diagnostic yield of testing sequential stools was assessed during two hospital epidemics of Clostridium difficile. Using a rapid immunoassay, C. difficile-associated disease was diagnosed in 237 diarrhoeal patients, of whom 204 (86%) were diagnosed from the first faeces sample and 12 (5%) were diagnosed from follow-up samples obtained within 1 week. The remaining 21 (9%) patients yielded a positive test from stools obtained >1 week after the initial negative sample. It was concluded that repeated testing of stools for C. difficile toxin is of value in controlling outbreaks of C. difficile infection. PMID- 18393997 TI - spa typing directly from a mecA, spa and pvl multiplex PCR assay-a cost-effective improvement for methicillin-resistant Staphylococcus aureus surveillance. AB - Rapid detection and typing of methicillin-resistant Staphylococcus aureus are important components of infection control programmes. A protocol is described that enables sequencing of the spa gene fragment directly from a multiplex PCR targeting the clinically relevant mecA, pvl and spa genes, resulting in high throughput characterisation of S. aureus. Implementation of the method in the Danish national reference laboratory has markedly reduced the use of reagents and the requirement for hands-on time, and has also provided fast typing results. In addition, the method reduces the risk of sample mishandling. PMID- 18393998 TI - The allene oxide cyclase family of Arabidopsis thaliana: localization and cyclization. AB - Jasmonates are derived from oxygenated fatty acids (oxylipins) via the octadecanoid pathway and are characterized by a pentacyclic ring structure. They have regulatory functions as signaling molecules in plant development and adaptation to environmental stress. Recently, we solved the structure of allene oxide cyclase 2 (AOC2) of Arabidopsis thaliana, which is, together with the other three AOCs, a key enzyme in the biosynthesis of jasmonates, in that it releases the first cyclic and biologically active metabolite -- 12-oxo-phytodienoic acid (OPDA). On the basis of models for the bound substrate, 12,13(S)-epoxy 9(Z),11,15(Z)-octadecatrienoic acid, and the product, OPDA, we proposed that a conserved Glu promotes the reaction by anchimeric assistance. According to this hypothesis, the transition state with a pentadienyl carbocation and an oxyanion is stabilized by a strongly bound water molecule and favorable pi-pi interactions with aromatic residues in the cavity. Stereoselectivity results from steric restrictions to the necessary substrate isomerizations imposed by the protein environment. Here, site-directed mutagenesis was used to explore and verify the proposed reaction mechanism. In a comparative analysis of the AOC family from A. thaliana involving enzymatic characterization, in vitro import, and transient expression of AOC-enhanced green fluorescent protein fusion proteins for analysis of subcellular targeting, we demonstrate that all four AOC isoenzymes may contribute to jasmonate biosynthesis, as they are all located in chloroplasts and, in concert with the allene oxide synthase, they are all able to convert 13(S)-hydroperoxy-9(Z),11(E),15(Z)-octadecatrienoic acid into enantiomerically pure cis(+)-OPDA. PMID- 18393999 TI - Order within a mosaic distribution of mitochondrial c-type cytochrome biogenesis systems? AB - Mitochondrial cytochromes c and c(1) are present in all eukaryotes that use oxygen as the terminal electron acceptor in the respiratory chain. Maturation of c-type cytochromes requires covalent attachment of the heme cofactor to the protein, and there are at least five distinct biogenesis systems that catalyze this post-translational modification in different organisms and organelles. In this study, we use biochemical data, comparative genomic and structural bioinformatics investigations to provide a holistic view of mitochondrial c-type cytochrome biogenesis and its evolution. There are three pathways for mitochondrial c-type cytochrome maturation, only one of which is present in prokaryotes. We analyze the evolutionary distribution of these biogenesis systems, which include the Ccm system (System I) and the enzyme heme lyase (System III). We conclude that heme lyase evolved once and, in many lineages, replaced the multicomponent Ccm system (present in the proto-mitochondrial endosymbiont), probably as a consequence of lateral gene transfer. We find no evidence of a System III precursor in prokaryotes, and argue that System III is incompatible with multi-heme cytochromes common to bacteria, but absent from eukaryotes. The evolution of the eukaryotic-specific protein heme lyase is strikingly unusual, given that this protein provides a function (thioether bond formation) that is also ubiquitous in prokaryotes. The absence of any known c type cytochrome biogenesis system from the sequenced genomes of various trypanosome species indicates the presence of a third distinct mitochondrial pathway. Interestingly, this system attaches heme to mitochondrial cytochromes c that contain only one cysteine residue, rather than the usual two, within the heme-binding motif. The isolation of single-cysteine-containing mitochondrial cytochromes c from free-living kinetoplastids, Euglena and the marine flagellate Diplonema papillatum suggests that this unique form of heme attachment is restricted to, but conserved throughout, the protist phylum Euglenozoa. PMID- 18394000 TI - Adverse drug reaction driven immunosuppressive drug manipulations: a single center comparison of enteric-coated mycophenolate sodium vs. mycophenolate mofetil. AB - Enteric-coated mycophenolate sodium (MPS) has been developed to help circumvent the upper gastrointestinal side-effects of mycophenolic acid by facilitating drug release in the small intestine. Many questions regarding the side-effect profile of MPS remain. Therefore, the purpose of this study is to review a single center's experience with mycophenolate mofetil (MMF) and MPS. METHODS: This retrospective, sequential cohort analysis of de novo renal and pancreas transplants (n = 198) compared MMF 500 mg b.i.d. to MPS 360 mg b.i.d. in conjunction with antibody induction, tacrolimus, and steroids. RESULTS: There were fewer adverse event driven drug manipulations in the MPS group at 90 d (4% MPS vs. 17% MMF) and 180 d (10% MPS vs. 24% MMF, p = 0.006, log-rank) after transplantation. There was a trend toward fewer GI-related hospital admissions in the MPS arm (7% MPS vs. 13% MMF, p = 0.18). Allograft outcomes including patient survival, graft survival, acute rejection, serum creatinine, and infection were similar. CONCLUSION: This single-center, sequential cohort study demonstrates that MPS is associated with fewer adverse event driven drug manipulations while maintaining similar safety and allograft outcomes. PMID- 18394001 TI - Pharmacodynamic immune monitoring of NFAT-regulated genes predicts skin cancer in elderly long-term renal transplant recipients. AB - INTRODUCTION: Among elderly allograft recipients non-melanoma skin cancer (NMSC) is the most common malignancy. We have previously shown that malignancies are associated with a higher intensity of ciclosporin A (CsA)-induced immunosuppression. METHOD: Fifty-five long-term elderly renal transplant patients with a stable transplant function had regular skin examinations. The expression of the nuclear factor of activated T cells (NFAT)-regulated genes (interleukin-2, granulocyte-macrophage colony stimulating-factor, interferon-gamma) was determined by real-time PCR at CsA trough levels and two h after oral intake. RESULTS: The CsA dose was 2.0 mg/kg (0.95-3.50), with CsA trough level (C0) level 97 microg/L (33-157) and CsA two-h level (C2) 538 microg/L (350-1228). NMSC was diagnosed in 14/55 patients (25.4%). A total of 85.7% of allograft recipients with NMSC were male (p < 0.005). Age, time after transplantation, CsA dose, CsA C0 and C2 level were comparable in both groups. NFAT-regulated gene expression was signficantly lower in patients with skin cancer compared with patients without skin cancer [4.94% (0.91-13.4) vs. 11.6% (3.3-40.8), p < 0.001). CONCLUSION: The unproportional high incidence of NMSC in elderly long-term kidney transplanted patients correlates with a lower NFAT-regulated gene expression which is a surrogate biomarker for a higher degree of functional immunosuppression. Further studies are required to determine whether the reduction of CsA with an increased NFAT-regulated gene expression is associated with a lower NMSC incidence. PMID- 18394002 TI - Does circulating BNP normalize after heart transplantation in patients with normal hemodynamic and right and left heart functions? AB - BACKGROUND: Increased brain natriuretic peptide (BNP) in cardiovascular disease is thought to be a compensatory protective mechanism allowing to delay the occurrence of terminal heart failure. Heart transplantation should normalize the neuroendocrine balance but BNP remains elevated in stable heart-transplant recipients (Htx). Such increase has been related to persistent endothelial and cardiac dysfunctions. The purpose of this study was to determine whether selected Htx, presenting with normal hemodynamic and cardiac systolic and diastolic functions on both side of the heart, show a normalization of their BNP plasma values. METHODS: Of a cohort of well-being 26 Htx, we selected 12 patients with normal hemodynamics and left and right heart systolic and diastolic functions and compared their circulating BNP, cyclic guanosine monophosphate (cGMP) (the BNP second messenger) and endothelin-1 (ET) values with that of 12 age-, body mass index- and mean arterial pressure-matched controls. Cardiac function determination by echodoppler included cardiac filling pressures assessment using tissue Doppler imaging. Blood samples for biological and hormonal determinations were drawn at rest, within 15 min before echocardiography. RESULTS: As selected, hemodynamic and left and right heart systolic and diastolic functions were located in the normal range in Htx. Plasma ET value was also similar in Htx and controls (20.7 +/- 0.9 vs. 19.6 +/- 0.9 fmol/mL). However, circulating BNP, like cGMP, was still significantly increased after heart transplantation, when compared with controls (33.8 +/- 8.5 vs. 4.0 +/- 0.9 pg/mL, p = 0.002 and 8.2 +/- 1.1 vs. 4.4 +/- 0.3 nmol/L, p = 0.003) for BNP and cGMP, respectively, in Htx and controls. Interestingly, the sole correlation observed was between BNP and cGMP (r = 0.85, p < 0.0001) after heart transplantation. CONCLUSIONS: After heart transplantation, BNP remained increased despite the normalization of hemodynamic and cardiac systolic and diastolic functions. This suggests that such endocrine heart stimulation should not be viewed only as a hemodynamic marker in Htx. Further studies will be useful to investigate the role of pro-inflammatory cytokines and whether elevated BNP still possesses antifibrotic properties, further supporting the interest of enhancing its activity after heart transplantation. PMID- 18394003 TI - The ups and downs of holoprosencephaly: dorsal versus ventral patterning forces. AB - Holoprosencephaly (HPE), characterized by incomplete separation of forebrain and facial components into left and right sides, is a common developmental defect in humans. It is caused by both genetic and environmental factors and its severity covers a wide spectrum of phenotypes. The genetic interactions underlying inherited forms of HPE are complex and poorly understood. Animal models, in particular mouse mutants, are providing a growing understanding of how the forebrain develops and how the cerebral hemispheres become split into left and right sides. These insights, along with the characterization to date of some of the genes involved in human HPE, suggest that two distinct mechanisms underlie the major classes of HPE, 'classic' and midline interhemispheric (MIH). Disruption either directly or indirectly of the ventralizing effect of sonic hedgehog signaling appears central to all or most forms of classic HPE, while disruption of the dorsalizing effect of bone morphogenetic protein signaling may be key to cases of MIH HPE. PMID- 18394004 TI - First case of compound heterozygosity in ALS2 gene in infantile-onset ascending spastic paralysis with bulbar involvement. PMID- 18394005 TI - Updated growth curves for Turkish children aged 15 days to 60 months. AB - BACKGROUND: Growth reference values are useful in paediatric health care as a health indicator. Secular changes in height for age values are also known to affect the timing of puberty and brain weight. Different populations may be at different stages of this secular trend. It is, therefore, necessary to periodically update the growth reference values for each population to identify these changes. The aim of this study was to update the growth reference values for Turkish infants and young children. METHODS: Background information and height/length, weight and head circumference measurements on a sample of 2391 boys and 2102 girls were obtained from Well Child Clinic Records. The LMS method was used for the analyses. The percentage of exclusive breastfeeding was 62% at 4 months and 26.6% at 6 months. The continuation rate of breastfeeding at 12 months was 62.5%. RESULTS: Comparison with previous Turkish data showed an increase in height of 0.9 cm for boys and 1.1 cm for girls at age 5 years. The increase in weight was 0.220 kg in girls, while a decrease of 0.160 kg was noted in boys. Comparison with the 2000 US and Swedish values indicated that population differences may exist in pre-pubertal years. CONCLUSION: This study provides an enhanced instrument to evaluate the growth of Turkish infants and young children. PMID- 18394006 TI - What helps when it hurts: children's views on pain relief. AB - BACKGROUND: Previous studies have focused on children's views of sources of pain and only secondarily explored their views on pain-relief strategies. METHODS: An exploratory cross-sectional descriptive design and 'draw and write' technique were used to investigate what children think helps them when they have pain. RESULTS: The sample (n = 71) was comprised of 33% boys and 67% girls, with an age range of 4-16 years (mean +/- SD: 9.25 +/- 3.04). Four overarching themes were derived that were common to both the texts and drawings: 'People who help', 'What I do that helps', 'What other people do that helps' and 'Things that help'. Children also described their emotional reactions to pain or pain relief. Multiple themes were represented in most of the texts and drawings. There were few differences in the themes present in the children's texts and drawings based on developmental age and no differences based on gender. CONCLUSIONS: Children across the three developmental age groups view themselves as active agents in pain relief. Although less than half of the children described specific behaviours they had taken, almost all children indicated their active role as the central figure in relation to use of objects or the actions of others. PMID- 18394007 TI - The evaluation of tailored and web-based information for new fathers. AB - BACKGROUND: Information and advice on infant health and development have been shown to be well received by new parents and to lead to more confident and nurturing parenting. However, in spite of the accumulating evidence highlighting the importance of fathers to the well-being of their families, fathers rarely access and utilize 'parenting' information. Tailored information for fathers delivered via email and Internet may provide an alternate route for support for fathers. This study aimed to assess father's readiness to utilize electronic information tailored to father's role. METHODS: Two hundred and fifty-three fathers from antenatal classes in two Australian cities were offered email and web-based information on seven topics: baby play, breastfeeding, post-natal depression, father-infant bonding, sex after birth, work-family balance and fussy babies. Of 137 who completed consent forms, a total of 105 fathers selected three topics and 67 fathers provided 149 topic evaluations. RESULTS: Most respondents were from managerial, professional or skilled occupations and had higher educational qualifications than the general population of fathers. The most popular topics were those that related to father-infant interaction (baby games and father-infant bonding), and the least popular were breastfeeding and sex after the birth. Respondents rated the information as satisfactory and most (78%) indicated that it changed their approach to fathering. Although the websites provided were available through the Internet, few fathers had previously accessed them. CONCLUSIONS: Information tailored to new father's perspectives provided through email or Internet may increase new fathers' access to useful knowledge and support. Important health topics such as breastfeeding may not currently be perceived as particularly relevant to fathers in the antenatal period. Portals designed to filter existing websites could facilitate the use of parenting information by new fathers. PMID- 18394008 TI - Three-layered structure shared between Lewy bodies and lewy neurites-three dimensional reconstruction of triple-labeled sections. AB - Lewy bodies (LBs) and Lewy neurites (LNs) are the hallmarks of Parkinson's disease (PD). Although LBs and LNs, frequently coexistent, share some histological properties, their appearances are quite different under conventional two-dimensional observation. In order to clarify how these apparently different structures (LBs and LNs) are related during their formation, we performed three dimensional observation on post-mortem brainstem tissues with PD. Sixty-microm thick floating sections were multi-immunofluorolabeled for alpha-synuclein (alphaS), ubiquitin (Ub) and neurofilament (NF). Serial confocal images were reconstructed with software. External three-dimensional configuration of LBs, double-labeled for alphaS and NF, exhibited frequent continuity with LNs (70%). Internally, alphaS and Ub formed the three-dimensional concentric inner layers and NF rimmed these inner layers. This layered structure was shared among spherical LBs, rod-shaped LNs and even convoluted forms of LBs/LNs. Furthermore, each layer exhibited continuity without interruption even in the convoluted form and around its junction to spherical LBs. This three-layered structure shared among various Lewy pathologies and their layered continuity on three-dimensional basis favor the hypothesis that LNs evolve into LBs. Besides progression from pale bodies to LBs, structural evolution from LNs into LBs may provide an alternative explanation for the variability of alphaS deposits and their interrelation. PMID- 18394009 TI - Expression of integrin alphavbeta3 in gliomas correlates with tumor grade and is not restricted to tumor vasculature. AB - In malignant gliomas, the integrin adhesion receptors seem to play a key role for invasive growth and angiogenesis. However, there is still a controversy about the expression and the distribution of alpha(v)beta(3) integrin caused by malignancy. The aim of our study was to assess the extent and pattern of alpha(v)beta(3) integrin expression within primary glioblastomas (GBMs) compared with low-grade gliomas (LGGs). Tumor samples were immunostained for the detection of alpha(v)beta(3) integrin and quantified by an imaging software. The expression of alpha(v)beta(3) was found to be significantly higher in GBMs than in LGGs, whereby focal strong reactivity was restricted to GBMs only. Subsequent analysis revealed that not only endothelial cells but also, to a large extent, glial tumor cells contribute to the overall amount of alpha(v)beta(3) integrin in the tumors. To further analyze the integrin subunits, Western blots from histologic sections were performed, which demonstrated a significant difference in the expression of the beta(3) integrin subunit between GBMs and LGGs. The presented data lead to new insights in the pattern of alpha(v)beta(3) integrin in gliomas and are of relevance for the inhibition of alpha(v)beta(3) integrin with specific RGD peptides and interfering drugs to reduce angiogenesis and tumor growth. PMID- 18394010 TI - Hypoxia-inducible factor-2alpha: effect on radiation sensitivity and differential regulation by an mTOR inhibitor. AB - OBJECTIVE: To determine the role of hypoxia-inducible factor-2alpha (HIF2alpha) on the sensitivity of renal cell carcinoma (RCC) cell lines to ionizing radiation and to determine if the mTOR antagonist, rapamycin, could decrease HIF2alpha protein levels. MATERIALS AND METHODS: Cell lines expressing stable short-hairpin RNAs (shRNAs) encoding HIF2alpha shRNAs or an empty vector were transfected with a hypoxia responsive element (HRE)-driven firefly luciferase reporter gene. Two separate paired cell lines were assayed for their response to increasing doses of ionizing radiation. Proliferation and cell cycle kinetics were compared for cell lines expressing HIF2alpha shRNAs and empty vectors. The effect of an mTOR antagonist, rapamycin on HIF1alpha and HIF2alpha proteins levels was also assessed. RESULTS: We confirmed that the 786-O RCC lines with stably integrated shRNAs against HIF2alpha had decreased activation of a plasmid with a HRE-driven firefly luciferase reporter gene. Lines from two separate cell clones with decreased HIF2alpha levels showed a significant increase in radiation sensitivity and an increase in G2 cell cycle arrest. Rapamycin, while effective in decreasing HIF1alpha protein levels, did not affect HIF2alpha levels in either of the RCC cell lines. CONCLUSIONS: These results show that decreasing levels of HIF2alpha leads to an increased sensitivity to ionizing radiation. This finding may explain in part, the known resistance of RCC to radiation therapy. Although mTOR antagonists are approved for the treatment of RCC, these agents do not decrease HIF2alpha levels and therefore might not be effective in enhancing the radio sensitivity of these tumours. PMID- 18394011 TI - Functional and molecular identification of pH-sensitive K+ channels in murine urinary bladder smooth muscle. AB - OBJECTIVE: To examine the role of pH-sensitive K(+) channels in setting the resting membrane potential in murine bladder smooth muscle, as bladder contractility is influenced by the resting membrane potential, which is mainly regulated by background K(+) conductances. MATERIALS AND METHODS: Using conventional microelectrode recordings, isometric tension measurements, patch clamp recordings, reverse transcription-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry, we assessed bladder smooth muscle cells and tissues. RESULTS: Acidic pH (pH 6.5) depolarized the resting membrane potential of murine bladder smooth muscles and increased muscle tone and contractility. The pH-induced changes were not abolished by neuronal blockers or classical K(+)-channel antagonists. Lidocaine (1 mM) and bupivacaine (100 microm) mimicked the effects of acidifying the external solution, and in the presence of lidocaine no further increase in contractility was induced by reducing the pH to 6.5. Voltage-clamp experiments on freshly dispersed bladder myocytes showed that pH 6.5 decreased the outward current. Pre-treatment of bladder myocytes with the classical K(+) antagonists tetraethylammonium (10 mm), 4-aminopyridine (5 mM), glibenclamide (10 microm) or apamin (300 nM) did not inhibit the effects of low pH on outward current. However, treatment with lidocaine (1 mM) abolished the effects of acidic pH on outward current. RT-PCR showed the expression of the acid sensitive K(+) channel (TASK)-1 and TASK-2 gene transcripts in murine bladder, and immunohistochemistry and Western blot analysis showed TASK-1 and TASK-2 channel expression and distribution in smooth muscle tissues and cells. CONCLUSION: TASK channels are expressed in bladder smooth muscle and contribute to the basal K(+) conductances responsible for resting membrane potential. PMID- 18394012 TI - Systemic and bronchodilator effects of inhaled rac-formoterol in subjects with chronic obstructive pulmonary disease: a dose-response study. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: The long-acting inhaled beta(2)-agonist formoterol has systemic effects when taken in high doses. It can be used as relief medication in asthma and there is interest in this approach in chronic obstructive pulmonary disease (COPD). Relief medication can involve high doses, and in subjects with COPD who have limited ability to bronchodilate the adverse effects can outweigh the benefits. There are concerns with the overall safety of high-dose beta(2)-agonists in subjects with COPD, and this study looks at the balance of beneficial and adverse effects of a range of doses of inhaled formoterol. WHAT THIS STUDY ADDS: Among subjects with COPD, high-dose inhaled formoterol produced a dose-related increase in forced expiratory volume in 1 s without a corresponding reduction in dyspnoea or increase in walk distance. Systemic effects were modest, however, and high doses did not appear to reduce patient satisfaction. Although further safety data are needed, inhaled formoterol may have a role as relief medication in COPD. AIMS Rac-formoterol is effective as maintenance treatment for both asthma and chronic obstructive pulmonary disease (COPD) and is now used as relief therapy in asthma. Using rac-formoterol for relief and maintenance treatment could involve inhalation of high doses, and whether this is of benefit in COPD is uncertain. Our aim was to determine whether higher doses of inhaled rac-formoterol produce systemic adverse effects that outweigh the limited bronchodilator benefit seen in subjects with COPD. METHODS: We examined airway and systemic effects of 6, 12, 24 and 48 microg rac-formoterol and placebo on separate days in 20 subjects with symptomatic COPD [forced expiratory volume in 1 s (FEV(1)) 47% predicted]. FEV(1), oxygen saturation, dyspnoea, 6-min walk distance, patient satisfaction and systemic effects were measured and treatment was assessed against placebo and for dose-response effects. RESULTS: FEV(1)[area under the time-response curve (AUC)] and satisfaction scores increased with all formoterol doses compared with placebo, as did AUC tremor with the 24- and 48-microg doses and AUC heart rate with the 48 microg dose. A dose-response relationship was seen with FEV(1) and tremor, but not with satisfaction scores. There was no difference between placebo and rac formoterol for other variables. CONCLUSIONS: Our results show that in patients with COPD rac-formoterol improves FEV(1) and patient satisfaction without a corresponding reduction in dyspnoea. Since the systemic effects from a relatively high dose were minimal, its use as relief medication in COPD merits further evaluation. PMID- 18394013 TI - The importance of anticoagulant agents in measuring platelet aggregation in patients treated with clopidogrel and aspirin. PMID- 18394014 TI - Differential impact of conventional-dose and low-dose postmenopausal hormone therapy, tibolone and raloxifene on C-reactive protein and other inflammatory markers. AB - BACKGROUND: Postmenopausal hormone therapy (HT) is associated with an increased risk for arterial and venous thrombosis. OBJECTIVES: To compare the impact of HT, tibolone, and raloxifene on C-reactive protein (CRP) and other inflammatory markers, and to investigate possible underlying mechanisms for changes in CRP and D-dimer. METHODS: Two hundred and two healthy women were randomly assigned to treatment for 12 weeks with either low-dose HT containing 1 mg of 17beta estradiol and 0.5 mg of norethisterone acetate (NETA) (n = 50), conventional-dose HT containing 2 mg of 17beta-estradiol and 1 mg of NETA (n = 50), 2.5 mg of tibolone (n = 51), or 60 mg of raloxifene (n = 51). RESULTS: CRP increased in the conventional-dose HT and low-dose HT groups. These changes were significantly more pronounced in the conventional-dose HT group (RMANOVA, P = 0.02). Also, tibolone was associated with an increase in CRP, in contrast to raloxifene, which reduced CRP. Reductions in levels of Lp(a), intercellular adhesion molecule-1 (ICAM-1), P-selectin, E-selectin, monocyte chemotactic protein 1 (MCP-1) and interleukin-6 (IL-6) were observed in all treatment groups. The changes were most pronounced for the conventional-dose HT group, and least pronounced for the raloxifene group, whereas the changes in those allocated to tibolone and low-dose HT were intermediary. Increased levels of tumor necrosis factor (TNF)-alpha and von Willebrand factor (VWF) were seen in the raloxifene group. We observed positive associations between changes in IL-6, VWF, MCP-1, and CRP. CONCLUSIONS: The regimens had markedly different impacts on markers of inflammation. The average increase in CRP was not accompanied by increases in the average levels of IL-6, TNF-alpha or other markers, but women with large reductions in IL-6 had reduced increases in CRP. PMID- 18394015 TI - The role of 18F-FDOPA and 18F-FDG-PET in the management of malignant and multifocal phaeochromocytomas. AB - BACKGROUND: (18)F-DOPA has emerged as a promising tool in the localization of chromaffin-tissue-derived tumours. Interestingly, phaeochromocytomas (PHEO) are also FDG avid. AIM AND METHODS: The aim of this study was to retrospectively evaluate the results of (18)F-FDOPA and/or (18)F-FDG-PET in patients with PHEO and paragangliomas (PGLs) and to compare the outcome of this approach with the traditional therapeutic work-up. Nine patients with non-MEN2 related PHEO or PGL were evaluated. At the time of the PET studies, the patients were classified into three groups based on their clinical history, conventional and SPECT imaging. The groups were malignant disease (n = 5, 1 VHL), apparently unique tumour site in patients with previous surgery (n = 1, SDHB) and multifocal tumours (n = 3, 1 VHL, 1 SDHD). (18)F-FDOPA and (18)F-FDG-PET PET/CT were then performed in all patients. RESULTS: PET successfully identified additional tumour sites in five out of five patients with metastatic disease that had not been identified with SPECT + CI. Whilst tumour tracer uptake varied between patients it exhibited a consistently favourable residence time for delayed acquisitions. (18)F-FDOPA uptake (SUVmax) was superior to (18)F-FDG uptake in cases of neck PGL (three patients, four tumours). If only metastatic forms and abdominal PGLs were considered, (18)F-FDG provided additional information in three cases (two metastatic forms, one multifocal disease with SDHD mutation) compared to (18)F FDOPA. CONCLUSIONS: Our results suggest that tumour staging can be improved by combining (18)F-FDOPA and (18)F-FDG in the preoperative work-up of patients with abdominal and malignant PHEOs. (18)F-FDOPA is also an effective localization tool for neck PGLs. MIBG however, still has a role in these patients as MIBG and FDOPA images did not completely overlap. PMID- 18394016 TI - 123I-metaiodobenzylguanidine (MIBG) scintigraphy for the detection of adrenal and extra-adrenal phaeochromocytomas: CT and MRI correlation. AB - CONTEXT: Evidence regarding the accuracy of [(123)I] metaiodobenzylguanidine (MIBG) imaging for phaeochromocytoma localization is currently limited to small series. OBJECTIVE: We present the largest series of primary phaeochromocytomas in which the performance of [(123)I]MIBG has been evaluated and correlated with cross-sectional imaging. DESIGN: We identified 76 patients with both preoperative [(123)I]MIBG and cross-sectional imaging for confirmed primary phaeochromocytoma between 1995 and 2005 at our institution. This comprised 60 adrenal tumours in 55 patients and 33 extra-adrenal tumours in 23 patients (2 patients had both adrenal and extra-adrenal tumours). Phaeochromocytoma metastases were not evaluated. MAIN OUTCOME MEASURE(S): [(123)I]MIBG studies were independently reviewed and correlated with CT and MRI examinations, as well as tumour functional status, to identify features that may predict a false negative [(123)I]MIBG result. RESULTS: The overall sensitivity of [(123)I]MIBG was 75%. Tumour detection was lower for extra-adrenal (58%) vs. adrenal (85%) phaeochromocytomas (P = 0.005). For extra adrenal tumours, [(123)I]MIBG demonstrated 8 of 14 carotid body, 2 of 2 intrathoracic, 8 of 14 retroperitoneal and 2 of 3 pelvic phaeochromocytomas. Overall, MRI and CT demonstrated 68 of 68 and 72 of 74 primary phaeochromocytomas, respectively. Tumour size correlated with [(123)I]MIBG uptake for adrenal (P = 0.009) but not extra-adrenal tumours. When tumours were adjusted for size, no other imaging feature or functional status correlated with [(123)I]MIBG negativity, although two large [(123)I]MIBG negative adrenal tumours contained large areas of necrosis or haemorrhage. CONCLUSIONS: Extra-adrenal and small adrenal phaeochromocytomas are more likely to result in false negatives on [(123)I]MIBG. Tumoural necrosis or haemorrhage do not consistently relate to [(123)I]MIBG uptake, although adrenal phaeochromocytomas containing minimal solid tissue due to extensive necrosis may predict a negative [(123)I]MIBG result. PMID- 18394017 TI - New mutation type in pseudohypoparathyroidism type Ia. AB - CONTEXT: The GNAS gene encodes the alpha-subunit of the stimulatory G proteins, which play a crucial role in intracellular signal transduction of peptide and neurotransmitter receptors. Heterozygous inactivating maternally inherited mutations of GNAS (including translation initiation mutations, amino acid substitutions, nonsense mutations, splice site mutations and small insertions or deletions) lead to a phenotype in which Albright hereditary osteodystrophy is associated with pseudohypoparathyroidism type Ia. OBJECTIVE: We sought to identify the molecular defect in a patient who was thought to have PHP-Ia. METHODS AND RESULTS: The GNAS gene of a 5-year-old boy with brachydactily, mental retardation, pseudohypoparathyroidism and congenital hypothyroidism was investigated. We found a heterozygous inversion of exon 2 and part of intron 1 of de novo origin. Molecular studies of cDNA from blood RNA demonstrated that both the normal and the mutant variants were stable and that new splice-sites were generated. CONCLUSION: This report demonstrates the first evidence for an inversion at the GNAS gene responsible of pseudohypoparathyroidism type Ia. PMID- 18394018 TI - Pituitary incidentaloma. PMID- 18394019 TI - Thyrotoxicosis during sunitinib treatment for renal cell carcinoma. AB - CONTEXT: Sunitinib malate is an oral tyrosine kinase inhibitor used in the treatment of renal cell carcinoma (RCC) and gastrointestinal stromal tumours. Hypothyroidism has been observed in patients treated with sunitinib, but the mechanism whereby sunitinib induces hypothyroidism is unknown. OBJECTIVE: To describe a series of six patients who developed thyrotoxicosis while on sunitinib for metastatic RCC. SETTING: The study was conducted at Austin Health, a tertiary teaching hospital in Melbourne, Australia. RESULTS: Two patients developed severe thyrotoxicosis within 10 weeks after commencing sunitinib. In contrast, in the four patients who presented with later onset (16-30 weeks) thyrotoxicosis, the thyrotoxicosis was relatively mild, self-limiting and rapidly progressed to hypothyroidism. These patients experienced recurrent episodes of thyrotoxicosis in temporal relation to their cyclical sunitinib treatment. One patient had cytological evidence of lymphocytic thyroiditis. CONCLUSIONS: These findings suggest that sunitinib-induced hypothyroidism may be a consequence of preceding thyroiditis with associated transient thyrotoxicosis. As predictive factors are currently unknown, we suggest regular monitoring of thyroid function in all patients commenced on sunitinib. Clinicians treating patients with sunitinib or other similar kinase inhibitors should to be alerted to thyroid dysfunction as a potential toxicity of these agents. PMID- 18394020 TI - Evaluation of myocardial fibrosis by imaging techniques in acromegaly. PMID- 18394021 TI - Serum testosterone and bioavailable testosterone correlate with age and body size in hypogonadal men treated with testosterone undecanoate (1000 mg IM--Nebido). AB - OBJECTIVE: To investigate the loading regimen for intramuscular (IM) testosterone undecanoate (Nebido) to determine whether testosterone and bioavailable testosterone levels achieved correlate with age or body size of subjects studied. DESIGN: Retrospective observational study of testosterone naive patients and patients previously treated with an alternative testosterone therapy. PATIENTS: 51 hypogonadal men (35, 68.6% secondary hypogonadism). 8 (16%) had not previously received testosterone therapy. MEASUREMENTS: Patients received an IM injection of Nebido (1000 mg) at baseline and a second injection after 6 weeks. Serum was assayed at baseline and 18 weeks after commencing Nebido for total testosterone (TT) and SHBG. Bioavailable testosterone was calculated (cBioT) using TT and SHBG. Measurements were taken for weight, body mass index (BMI) and body surface area (BSA). RESULTS: Baseline TT (mean 11.5 nmol/l, range 0.3-54.8) increased by 50% after commencing Nebido (17.2 nmol/l (5.4-32.8), P = 0.0001). 75% of subjects had a TT within the reference range (8.0-25.0 nmol/l). Subjects with primary hypogonadism had a higher 18-week TT [20.9 nmol/l (9.8-32.8) vs. 15.5 (5.4-32.6), P = 0.02] and SHBG [39.2 nmol/l (11-82) vs. 25.7 (9.0-60.0), P = 0.003] although the cBioT was not significantly different [4.9 nmol/l (2.9-7.3) vs. 4.2 (2.0 7.9), P = 0.12]. The 18-week TT positively correlated with age (R = 0.36, P = 0.01) and negatively correlated with weight (R = -0.38, P = 0.006), BMI (R = 0.42, P = 0.002) and BSA (R =-0.38, P = 0.007). Similarly cBioT correlated with age (R = 0.28, P = 0.04), weight (R = -0.29, P = 0.03), BMI (R = -0.30, P = 0.03) and BSA (R = -0.27, P = 0.05). Age (t = 2.04, P = 0.05) and baseline testosterone (t = -9.26, P < 0.0001) were independent variables of the increase in TT at 18 weeks. CONCLUSION: This starting regimen is simple and provides the majority of men with a TT within the reference range. Age and baseline TT are independent variables of the increase in TT with IM testosterone undecanoate. At week 18 age and body size correlated with the cBioT and TT and this may then be used to estimate dosing frequency for this therapy. PMID- 18394022 TI - Lipid-lowering therapy does not affect the postprandial drop in high density lipoprotein-cholesterol (HDL-c) plasma levels in obese men with metabolic syndrome: a randomized double blind crossover trial. AB - INTRODUCTION: The postprandial lipid metabolism in metabolic syndrome patients is disturbed and may add to the increased cardiovascular risk in these patients. It is not known whether postprandial high density lipoprotein-cholesterol (HDL-c) metabolism is also affected and whether this can be influenced by statin and/or ezetimibe treatment. METHODS: Prospective, randomized, double blind, crossover trial comparing simvastatin 80 mg with simvastatin/ezetimibe 10 mg/10 mg treatment for 6 weeks on postprandial HDL-c metabolism in 15, nonsmoking, male, obese metabolic syndrome patients (Adult Treatment Panel III, ATPIII). Only study medication was allowed. HDL-c concentrations, cholesteryl ester transfer (CET), CET protein (CETP) mass and adiponectin were measured before and after oral fat loading. ClinicalTrials.gov NCT00189085. RESULTS: Plasma HDL-c levels remained stable during continuous fasting following an overnight fast. Pre-fat load HDL-c concentrations without treatment, after simvastatin and simvastatin/ezetimibe treatment were 1.15 +/- 0.04, 1.16 +/- 0.05 and 1.11 +/- 0.04 mmol/l. Fat load induced a 11% drop in HDL-c plasma levels; 1.02 +/- 0.05 mmol/l (P < 0.001) which was not affected by either therapy. Triglyceride levels during fat load were similar after both treatments. Total CET increased from 9.73 +/- 0.70 to 12.20 +/ 0.67 nmol/ml/h (P = 0.004). Four hours after fat loading CETP mass was increased while adiponectin levels were decreased, irrespective of treatment. DISCUSSION: HDL-c levels decrease as CET increases after fat loading in obese metabolic syndrome patients. This is not influenced by either simvastatin or simvastatin/ezetimibe treatment. After fat loading, CETP mass and CET increased, and adiponectin decreased pointing towards a potential role for intra-abdominal fat. Decreased postprandial HDL-c levels may contribute to the increased cardiovascular risk in metabolic syndrome patients on top of already low HDL-c levels. PMID- 18394023 TI - Anti-inflammatory effect of retinoic acid on prostaglandin synthesis in cultured cortical astrocytes. AB - Prostanoids are important mediators of inflammation and pain signaling. Although it is now well accepted that astrocytes participate in inflammatory reactions in the CNS, the molecular regulation of this activity is still largely unknown. Specifically, the regulation of prostanoid synthesis by this type of glia remains to be resolved. Recent evidence suggests that the transcriptional regulator retinoic acid (RA) is involved in regulation of the immune response. We have investigated the expression pattern of the enzymes that catalyze prostanoid and leukotriene synthesis in cultured cortical astrocytes, their stimulation by lipopolysaccharides (LPS) and their regulation by RA. The data indicate that astrocytes are an important source of prostaglandins (PGs) and that RA reduces their inflammatory biosynthesis. LPS treatment induced the expression of enzymes for the production of arachidonic acid and PGs but caused down-regulation of a PG degrading enzyme and of leukotriene synthesizing enzymes that compete with PG synthesis. Consequently, the secretion of the PGE(2) was highly increased after LPS exposure. RA counteracted the inflammatory regulation of cyclooxygenase (COX) 2 mRNA and protein in astrocytes and thereby reduced the synthesis of PGE(2) by approximately 60%. In the absence of LPS, RA enhanced the expression of COX-1 mRNA. In conclusion, RA might be effective in suppressing inflammatory processes in the brain by inhibiting PG synthesis. PMID- 18394024 TI - Potassium channels sensitive to combination of charybdotoxin and apamin regulate the tone of diabetic isolated canine coronary arteries. AB - AIMS: Functional roles of calcium-activated potassium channels on the mechanical activity of epicardial coronary arteries obtained from a canine model of diabetes were investigated. METHODS: Coronary arteries were isolated from healthy, alloxan diabetic and insulin-treated diabetic dogs. Basal tensions, contractions induced by the prostaglandin (PG) analogue, U46619, and endothelium-dependent relaxations to acetylcholine (ACh) were modified with charybdotoxin (CHTX) + apamin (APA), inhibitors of calcium-activated potassium channels, as well as with N(omega) nitro-l-arginine (LNA) + indomethacin (INDO) to suppress the synthesis of nitric oxide (NO) and PGs. The relaxing effect of nitroprusside-sodium (SNP), an NO donor, was also determined. RESULTS: In diabetic coronary arteries, CHTX + APA did not change while LNA + INDO elevated the basal tension. PG-induced contractions were enhanced by CHTX + APA and by LNA + INDO in all the three groups of animals. CHTX + APA decreased the maximal relaxations to ACh in a partly insulin-dependent manner. LNA + INDO abolished the endothelium-dependent relaxations to ACh. In diabetic coronary arteries, the sensitivity to SNP-induced relaxation was enhanced, insulin independently, suggesting that NO could be partly responsible for maintaining intact ACh-induced vasorelaxation. CONCLUSION: In diabetic canine coronary artery, the vasomotor responses reflect up-regulation of calcium-activated potassium channels. This endothelial mechanism of the canine epicardial coronary artery may oppose vasoconstrictions in diabetic vascular tissue. PMID- 18394025 TI - Nitric oxide depresses connexin 43 after myocardial infarction in mice. AB - AIMS: Heart failure (HF) is a major cause of death and morbidity. Connexin 43 (Cx43) content is reduced in the failing myocardium, but regulating factors have not been identified. In HF, inducible nitric oxide synthase (iNOS)-induced high levels of nitric oxide (NO) cause apoptosis and cardiac dysfunction. However, a direct iNOS-Cx43 link has not been demonstrated. We investigated this relationship in mice after myocardial infarction. METHODS: Effects of myocardial infarction were evaluated 2 weeks after coronary artery ligation in wild-type C57BL/6 (WT) and iNOS(-/-) knockout mice. Myocardial Cx43 and Cx45 content were assessed by immunofluorescence confocal imaging and western blotting. Cardiac function was evaluated in anaesthetized mice using a micro pressure-tipped catheter inserted into the left ventricle. RESULTS: Despite similar infarct size, deficiency in iNOS resulted in significantly lower plasma nitrate/nitrite levels, better haemodynamic performance and lower mortality 2 weeks after coronary ligation. Myocardial Cx43, but not Cx45, content was lower in WT mice following ligation. The reduction in Cx43 was less in iNOS(-/-) compared with WT mice. To assess the direct effect of NO on Cx43 expression, cultured neonatal mouse cardiomyocytes were employed. Incubation with the NO donor, S-nitroso-N acetylpenicillamine, elicited a dose-dependent decrease in Cx43 content in cultured neonatal cardiomyocytes. CONCLUSIONS: Increased NO production from iNOS depressed cardiac performance and contributed to the decreased myocardial Cx43 content 2 weeks after myocardial infarction. PMID- 18394027 TI - Seeing the person. PMID- 18394026 TI - Effect of medium- and long-chain fatty acid diets on PPAR and SREBP-1 expression and glucose homeostasis in ACBP-overexpressing transgenic rats. AB - AIM: Acyl-CoAs are important intermediates and regulators of lipid metabolism. Binding proteins like acyl-CoA binding protein (ACBP) can influence their regulatory functions. ACBP has also been shown to exert direct effects on gene regulation in vitro. As the physiological relevance of ACBP in the regulation of lipid metabolism under high fat diets is unclear, we investigated the influence of such diets on the metabolic responses in ACBP-overexpressing rats. METHODS: A transgenic rat line overexpressing the ACBP gene was used to study the effects of 4 weeks of feeding with medium- (MC) or long-chain (LC) fatty acid-containing diets. Glucose tolerance tests were performed. Expression of transcription factors was measured by quantitative RT-PCR and protein levels of AMP-activated protein kinase were determined by western blotting. RESULTS: Transgenic animals fed the MC diet had an improved glucose tolerance and lower serum insulin levels compared with controls. Their liver PPARgamma (by 43%) and SREBP-1 (by 35%) mRNA levels were found to be decreased, while adipose tissue PPARgamma expression was increased by 31%. Tg animals fed the LC diet did not exhibit changes in glucose or insulin levels but exhibited increased mRNA levels of liver PPARs and SREBP-1 (1.5-3.5 times) and decreased protein levels of AMPKalpha (by 48%). CONCLUSIONS: Our results demonstrate that ACBP overexpression affects metabolic responses to diets with distinct difference in their fatty acid chain lengths. The molecular regulatory mechanism behind these effects seems to be an ACBP-induced tissue specific regulation of expression of PPARs and SREBP. PMID- 18394028 TI - Parent-adolescent communication and diabetes: an integrative review. AB - AIM: This paper presents a synthesis of findings from empirical studies about communication and interaction between parents and their adolescents with diabetes. BACKGROUND: Communication between parent and adolescent is essential in transitioning the adolescent to increased responsibility for diabetes self management. Nurses are in a pivotal position to enhance the type of parent adolescent communication that facilitates this transition. DATA SOURCES: A search of published studies from 1985 to 2006 featuring communication or interaction between parents and adolescents with type 1 diabetes was implemented through a computerized search. REVIEW METHODS: Papers were organized by type of evidence and were analysed sequentially. Sources were described based on data elements which were extracted. Findings in the areas of productive and problematic parent adolescent communications were included. Data elements were compared and critiqued, noting consistencies, and findings were summarized and evaluated. RESULTS: Studies across several countries indicate that maternal support, conflict, control, involvement and emotional expression are important communication concepts that are linked to diabetes outcomes in adolescents. The influences of different family structures and cultural and socioeconomic circumstances, as well as developmental status and gender of adolescents on these types of communication, have not been systematically studied. CONCLUSION: Nurses caring for adolescents with type 1 diabetes need to consider family relationships and communication patterns in achieving health outcomes. Studies of communication, including perspectives of mothers and fathers, and the influence of family structure, economics and culture are needed to build a framework of parent-adolescent interaction and health outcomes for adolescents with type 1 diabetes. PMID- 18394029 TI - Governmentality, student autonomy and nurse education. AB - AIM: This paper is a report of a study to explore how governmental practices operated in nurse education. Background. Since the 1980s nurse education internationally has been strongly influenced by educational theories that aim to promote student autonomy by encouraging self-direction and critical thinking. Newer curriculum models advocate transformative approaches leading to greater emancipation, social equity and inclusion. Although these changes have been positively evaluated there had been limited critical research on how student behaviour is governed. METHOD: A discourse analytic study was conducted from 2000 to 2004 using interviews (n = 30) with a purposive sample students and teachers in one United Kingdom university. Data were also collated from the course curriculum and student handbook for the students' programme. Data were analysed to identify how student behaviour is governed. FINDINGS: Two governing practices are described: control and technologies of the self. These practices contribute to an overall system of governing student behaviour that creates tension between the avowed progressive empowerment discourse and taken for granted everyday educational practices. Students are subjected to a range of governmental and disciplinary strategies and, through a process of normalization, ultimately become their own supervisors within the system. CONCLUSION: The tensions between the demands of a professional outcome-based nursing programme and notions of empowerment and student autonomy have not been resolved. Instead, present educational practice is characterized by normalizing discursive practices that aim to produce a specific version of a student subject as autonomous learner. Thus, discourses of both empowerment and professional behaviour govern students. PMID- 18394030 TI - Living with a long-term urinary catheter: older people's experiences. AB - AIM: This paper is a report of a study of the experiences of older people living with a long-term urinary catheter and the development of a substantive theory grounded in their realities. BACKGROUND: Understanding the user perspective is a prerequisite for involving older people in a collaborative relationship with healthcare professionals which is built around their needs and wishes. Older people's views and experiences of the complexity of living with a long-term catheter have not been widely researched. An enhanced understanding of catheter users' perspectives and an awareness of their needs can be used to improve care. METHOD: A grounded theory approach was adopted and 20 in-depth interviews were carried out in England in 2005 and 2006 with 13 older people living at home with long-term urinary catheters. FINDINGS: The core category 'all about acceptance' described older people's adjustment to living with a long-term urinary catheter; the two categories 'at ease' and 'uneasy' reflect the extremes of contentment experienced. Three interlinking categories of 'trying to understand', 'judging catheter performance' and 'being aware of the catheter' shaped older people's relationships with their catheters and this was mediated by their 'interaction with others'. The consequences for older people fluctuated along a continuum from 'engaging actively' to experiencing 'downbeat sentiments'. CONCLUSION: To assist older people to adjust to living with a catheter, healthcare professionals must be sensitive to their life situations and individual needs rather than focusing predominantly on catheter performance and complications. PMID- 18394031 TI - Practice nurses and the facilitation of self-management in primary care. AB - AIM: This paper is a report of a study to explore practice nurse involvement in facilitation of self-management for long-term conditions. BACKGROUND: In the United Kingdom chronic disease services have shifted from secondary care to general practice and from general practitioners to practice nurses. A new United Kingdom General Practice contract requires adherence to chronic disease management protocols, and facilitating self-management is recognized as an important component. However, improving self-management is a relatively new focus and little is known about the ways in which nurses engage with patient self management and how they view work with patients in chronic disease clinics. METHOD: Semi-structured interviews with 25 practice nurses were carried out in 2004-2005. Interviews were audio-taped and transcribed verbatim. Analysis was informed by the 'trajectory model' and 'personal construct' theories. FINDINGS: Main themes in the early stages of work with patients were: categorization of patients, diagnosis, and patient education. First impressions appeared to determine expectations of self-management abilities, although these were amenable to change. Intermediate stages were 'ways of working' (breaking the task down, cognitive restructuring and addressing dissonance, modelling 'good' behaviour, encouragement, listening, involving carers and referral) and maintaining relationships with patients. However, in the longer-term nurses seemed to lack resources beyond personal experience and intuitive ways of working for encouraging effective self-care. CONCLUSION: The ways of working identified are unlikely to be sufficient to support patients' self-management, pointing to a need for education to equip nurses with techniques to work effectively with patients dealing with longer-term effects of chronic illness. PMID- 18394032 TI - How people with motor neurone disease talk about living with their illness: a narrative study. AB - AIM: This paper is a report of a study which explores patients' experiences and how they talk about living and coping with motor neurone disease. BACKGROUND: Living with motor neurone disease is challenging, frightening and disabling. It leads to progressive physical decline, normally with a prognosis of 3-5 years. Patients have to deal with many problems, including loss of mobility and the inability to communicate. There is little evidence about how people manage these problems or live with this illness. METHOD: Narrative case studies were used, the unit of analysis being a patient living in their own home or a care home. Thirteen adults were recruited through purposeful sampling. Longitudinal narrative interviews were conducted at three-monthly intervals over an 18-month period in 2005-06. Interviews were analysed focusing on the form and content of the patients' narratives. FINDINGS: Four types of narrative, or storyline were identified. The sustaining storyline is about living life as well as possible through keeping active and engaged in life. In contrast, the enduring storyline concerns living in an insurmountable situation leaving the person feeling disempowered, unable to fight for life or against death. Survival is the essence of the preserving storyline, while the fracturing storyline concerns loss and fear of what is to come. CONCLUSION: Storylines help make sense of complex narratives by encouraging closer attention and active listening to the stories and serve as organizing threads to help patients, families and healthcare professionals better understand living with motor neurone disease. PMID- 18394033 TI - Nurses' perceptions of evidence-based nursing practice. AB - AIM: This paper is a report of a study to investigate Registered Nurses' perceptions, attitudes and knowledge/skills associated with evidence-based practice. BACKGROUND: Evidence-based practice has emerged as a marker for healthcare quality. Previous studies have primarily used researcher-developed descriptive surveys to examine nurses' perceptions, as well as facilitators and barriers, related to evidence-based practice. Research suggests the value of understanding the organizational context prior to taking steps to implement evidence-based practice. METHODS: This study, conducted in 2006, had a descriptive, cross-sectional survey design using a psychometrically-validated measure of evidence-based practice. All Registered Nurses (n = 1031) employed by a large medical centre in the United States of America were asked to complete the questionnaires. The final response rate was 40.9% (n = 422). FINDINGS: Participants had moderate scores on practice and attitudes towards evidence-based practice. The knowledge/skills mean scores were somewhat lower. Statistically significant differences were found for attitudes between those with baccalaureate and higher education compared to those with associate and diploma education. The two most cited barriers to implementing evidence-based practice were time and knowledge. CONCLUSION: The findings suggest the value of a methodical assessment when developing a systematic plan for implementing an institutional culture of evidence-based practice. PMID- 18394034 TI - Multi-faceted palliative care intervention: aged care nurses' and care assistants' perceptions and experiences. AB - AIM: This paper is a report of a study to describe residential aged care nurses' and care assistants' perceptions of a multi-faceted palliative care intervention to identify potential areas to be addressed during subsequent action research phases. BACKGROUND: Action research was used to enhance the delivery of a palliative approach in residential aged care. The chronic care model guided the development of a multi-faceted intervention. This involved the: (1) establishment of a 'link nurse' role; (2) learning and development strategies for nurses, care assistants and general practitioners; (3) use of multi-disciplinary team meetings; and (4) access to specialist consultation. METHOD: A purposive sample (n = 28) of aged care nurses and care assistants participated in a series of four focus groups conducted in July 2005. Thematic content analysis of the transcripts was performed. FINDINGS: Four themes emerged: (1) targeted education can make a difference; (2) a team approach is valued; (3) clinical assessment tools are helpful; and (4) using the right language is essential. Participants described increased understanding of palliative care concepts, enhanced competencies, greater confidence to deliver palliative care and a desire to adopt a multi disciplinary approach to care planning. CONCLUSION: Sustaining a culture that is committed to ongoing learning and development interventions and creating multi disciplinary teams in the aged care setting is critical to embedding a palliative approach. The chronic care model is a useful framework to guide the development of interventions leading to better palliative care outcomes for residents and their families. PMID- 18394035 TI - Combining individual interviews and focus groups to enhance data richness. AB - AIM: This paper is a presentation of the critical reflection on the types of findings obtained from the combination of individual interviews and focus groups, and how such triangulation contributes to knowledge production and synthesis. BACKGROUND: Increasingly, qualitative method triangulation is advocated as a strategy to achieve more comprehensive understandings of phenomena. Although ontological and epistemological issues pertaining to triangulation are a topic of debate, more practical discussions are needed on its potential contributions, such as enhanced data richness and depth of inquiry. METHOD: Data gathered through individual interviews and focus groups from a study on patterns of cancer information-seeking behaviour are used to exemplify the added-value but also the challenges of relying on methods combination. FINDINGS: The integration of focus group and individual interview data made three main contributions: a productive iterative process whereby an initial model of the phenomenon guided the exploration of individual accounts and successive individual data further enriched the conceptualisation of the phenomenon; identification of the individual and contextual circumstances surrounding the phenomenon, which added to the interpretation of the structure of the phenomenon; and convergence of the central characteristics of the phenomenon across focus groups and individual interviews, which enhanced trustworthiness of findings. CONCLUSION: Although the use of triangulation is promising, more work is needed to identify the added value or various outcomes pertaining to method combination and data integration. PMID- 18394036 TI - Development and validation of a brief Occupational Coping Self-Efficacy Questionnaire for Nurses. AB - AIM: This paper is a report of a study to develop and test the psychometric properties of the Occupational Coping Self-Efficacy for Nurses Scale. BACKGROUND: Coping self-efficacy beliefs are defined as self-appraisals of capabilities to cope with environmental demands. People with higher levels of coping self efficacy beliefs tend to approach challenging situations in an active and persistent way, whereas those with lower levels of coping self-efficacy beliefs tend to direct greater energy to managing increasing emotional distress. METHOD: In 2006, 1383 nurses completed the following measures: Occupational Coping Self Efficacy Questionnaire for Nurses, Coping Inventory for Stressful Situations Short Form and Maslach Burnout Inventory. Based on a randomized split of the data, we conducted exploratory factor analysis on group 1 data (n = 691) and confirmatory factor analysis within the framework of structural equation modelling on group 2 data (n = 692). FINDINGS: The exploratory results revealed two factors: Coping Self-Efficacy to cope with the occupational burden (Cronbach alpha = 0.77) and Coping Self-Efficacy to cope with the relational burden (alpha = 0.79). In the confirmatory group, the two-factor structure was tested against an alternative one-factor structure and confirmed as the best solution. Correlation patterns between the Occupational Coping Self-Efficacy for Nurses Scales, and both coping and burnout variables, supported the criterion-related validity of the Occupational Coping Self-Efficacy for Nurses dimensions. CONCLUSION: Nurses can have two basic and distinct coping self-efficacy beliefs: beliefs about occupational burden and beliefs about relational difficulties in the workplace. Research is needed into how efficacy evaluations shift as a result of specific stress management interventions. PMID- 18394037 TI - How ethical is ethical research? Recruiting marginalized, vulnerable groups into health services research. AB - AIM: This paper is a discussion of how tensions in research ethics guidelines can have an impact on the recruitment of marginalized, vulnerable groups of people into health services research. Background. Research ethics guidelines in the United Kingdom (UK) predominantly reflect requirements for clinical trials and medical research but apply to all types of research conducted in the National Health Service. Guidelines emphasize the process of informed consent to protect individuals' autonomy and rights, and avoid harm to vulnerable people. This focus, however, can result in the exclusion of and discrimination against marginalized groups at a time when user involvement in research and health service developments is increasingly emphasized in policy. METHOD: This paper is based on a critical reflection on tensions observed in current research ethics guidelines in the UK. These tensions became apparent following recruitment of nine postnatal drug-using women to a qualitative phase of an evaluation in 2005 of a specialist health visitor service. FINDINGS: The main ethical tensions identified concerned the sampling and recruitment of marginalized groups, which can affect the validity and reliability of research findings. Consideration needs to be given to sampling and accessing such groups and assessing the capacity of individuals to give informed consent. This is to avoid coercion and exploitation of vulnerable individuals and groups in research, increase validity and reliability, and avoid pre-emptive exclusion of such groups in the research design. CONCLUSION: Ethical judgements and decisions about research can be contradictory and be influenced by competing paradigms. Use of the concept of 'responsible advocacy', with due consideration to the principles of autonomy, beneficence, non-maleficence and justice, is recommended. PMID- 18394038 TI - Power and empowerment in nursing: a fourth theoretical approach. AB - AIM: This paper is a discussion of the use of poststructuralism as a means of exploring power and empowerment in nursing. BACKGROUND: Power and empowerment are well-researched areas of nursing practice, but the issue of how to empower nurses and patients continues to cause debate. Power and empowerment are complex issues and other researchers have provided some clarity by proposing three theoretical approaches: critical social theory, organizational theory and social psychological theory. We support their work and propose an additional poststructural approach as a means of analyzing power and empowerment in nursing. DISCUSSION: The concept of power in nursing may be critiqued by drawing on the work of Michel Foucault and paying particular attention to two areas: disciplinary power and knowledge/power relationships. Foucault's contention was that behaviour is standardized through disciplinary power and that power and knowledge are intertwined. Nurses who seek an understanding of empowerment must first grasp such workings as hierarchical observation, normalizing judgement, the examination, and power/knowledge relationships, and that cognizance of such issues can promote nursing practice that is empowering. They need to adopt a more critical stance to understanding power and empowerment in nursing, and one way of fostering such criticism is to view nursing practice through a poststructural lens. CONCLUSION: A poststructural approach merits a place alongside other approaches to understanding power and empowerment in nursing. PMID- 18394039 TI - In response to Spillane V., Byrne M.C., Byrne M., Leathem C.S., O'Malley M. & Cupples M.E. (2007) Monitoring treatment fidelity in a randomized trial of a complex intervention. Journal of Advanced Nursing 60(3), 343-352. Important considerations for standardizing complex interventions. PMID- 18394040 TI - In response to: Paley J., Cheyne H., Dalgleish L., Duncan E. & Niven C. (2007) Nursing's ways of knowing and the dual process theories of cognition. Journal of Advanced Nursing 60(6), 692-701. Evidence for and evidence against. PMID- 18394041 TI - Evidence for Nursing Practice Updates from the Joanna Briggs Institute. Summaries of Systematic Reviews on nursing-related issues from the Cochrane Library, Joanna Briggs Institute and other evidence review organizations. PMID- 18394042 TI - Evidence for Nursing Practice Updates from the Joanna Briggs Institute. Summaries of Systematic Reviews on nursing-related issues from the Cochrane Library, Joanna Briggs Institute and other evidence review organizations. PMID- 18394044 TI - Treatment of progressive encephalomyelitis with rigidity and myoclonic jerks with rituximab: a case report. PMID- 18394045 TI - Creutzfeldt-Jakob disease: hopes for therapy. PMID- 18394046 TI - Functional electrical stimulation in neurological disorders. AB - Functional electrical stimulation (FES) refers to electrical stimulation of muscles in order to improve the impaired motor function. This is achieved by activating skeletal muscles with constant frequency trains of stimulations. This method has been found useful in various neurological disorders like hemiplegia, foot drop and paraplegia including spinal cord injuries. The first half of this review focuses on the broad clinical applications of functional electrical stimulation, its mechanism of action and the complications of this mode of therapy. Advanced Parkinson's disease (PD) is characterized by marked slowing of gait and frequent freezing episodes. Medical and surgical treatments are often ineffective in managing freezing episodes. The second half of this review discusses briefly the gait abnormalities in PD and the available treatment options. The possible role of FES in improving gait in parkinsonism and the importance of future research in this direction are highlighted. PMID- 18394048 TI - Effect of levetiracetam on cortical excitability: a transcranial magnetic stimulation study. AB - BACKGROUND AND PURPOSE: We studied the effect of levetiracetam (LEV), an anticonvulsant with a novel mechanism of action, on cortical excitability, measured using transcranial magnetic stimulation (TMS). For this purpose, 38 healthy volunteers were assessed in two TMS sessions, before and after an oral dose of 3000 mg LEV. METHODS: Resting motor threshold (RMT), intracortical facilitation (ICF) and intracortical inhibition (ICI), cortical silent period (CSP) threshold and duration and motor-evoked potential (MEP) amplitude were calculated. RESULTS: After treatment with LEV, RMT was increased (mean +/- SD: 63 +/- 14% of the maximum stimulator output) compared with baseline (58 +/- 11%). CSP threshold was decreased after LEV (54 +/- 10%; baseline, 57 +/- 11%). CSP duration was increased after LEV (116 +/- 37 ms; baseline: 102 +/- 33 ms). LEV did not affect ICF or ICI or mean MEP amplitude significantly. CONCLUSIONS: Our results indicate that LEV modulates some aspects of cortical excitability. Whereas the increase in the RMT most probably reflects the effect of LEV on ion channel activity, effects on the CSP might represent a modulation of GABA receptors at cortical and spinal level. PMID- 18394047 TI - Polymorphism in the sorbin and SH3-domain-containing-1 (SORBS1) gene and the risk of brain infarction in the Japanese population: the Fukuoka Stroke Registry and the Hisayama study. AB - BACKGROUND AND PURPOSE: Sorbin and SH3-domain-containing-1 (SORBS1) is an important adaptor protein in insulin-signalling pathway, and its genetic polymorphism may regulate the activity of insulin resistance. We investigated the association between the SORBS1 T228A polymorphism and ischaemic stroke. METHODS: Genotyping was achieved by a rapid-cycle PCR and melting curve analysis using fluorescent probes in 1049 incident cases of ischaemic stroke and 1049 age- and sex-matched control subjects recruited from the Hisayama study. RESULTS: The allele distributions of the SORBS1 T228A polymorphism were similar amongst cases and controls. The multivariate-adjusted odds ratio (OR) of the AA genotype for ischaemic stroke was 2.897 (95% CI, 0.907-8.018) compared with the TT genotype. In terms of stroke subtype, there was a trend toward a difference in the AA genotypes for lacunar infarction, compared with the TT genotype (OR = 8.740, P = 0.0510), and combined TT and TA genotypes (OR = 8.768, P = 0.0505). The other polymorphisms genotyped were not associated with any subtypes of ischaemic stroke. T228A polymorphism of SORBS1 was not associated with the prevalence of diabetes. CONCLUSIONS: The AA genotype of SORBS1 T228A polymorphism may play a role in lacunar infarction in the Japanese population. PMID- 18394049 TI - Refinement of the SPG9 locus on chromosome 10q23.3-24.2 and exclusion of candidate genes. AB - BACKGROUND AND PURPOSE: The hereditary spastic paraplegias (HSPs) are a heterogeneous group of neurodegenerative disorders, characterized by a progressive spasticity of the lower limbs. So far, 33 different loci (SPGs) have been mapped and the 15 genes responsible have been identified. We mapped a locus responsible for a form of spastic paraplegia, complicated by bilateral cataracts, gastroesophageal reflux with persisting vomiting and amyotrophy to chromosome 10q23.3-q24.2, in an Italian family. The critical region was in a 12 cm chromosomal interval between markers D10S564 and D10S603 (SPG9, MIM601162). In the same region, two other forms of HSP have been recently mapped: SPG27 and SPG33. In the latter case, the gene responsible has been identified. MATERIALS AND METHODS: To better characterize this region, we genotyped individuals from SPG9-linked families using additional markers and reduced the candidate region to a 4.8 Mb, excluding several genes by positional cloning. RESULTS: The refined SPG9 locus is positioned completely within SPG27 and does not include the SPG33 gene. DISCUSSION: Fifty-two transcripts are present in the refined critical region and 25 strong candidates have been excluded as disease causing genes by direct sequencing. Six of them were also excluded as responsible for SPG27. PMID- 18394051 TI - History of hemodialyzers' designs. AB - Accumulation of knowledge requisite for development of hemodialysis started in antiquity and continued through Middle Ages until the 20th century. Firstly, it was determined that the kidneys produce urine containing toxic substances that accumulate in the body if the kidneys fail to function properly; secondly, it was necessary to discover the process of diffusion and dialysis; thirdly, it was necessary to develop a safe method to prevent clotting in the extracorporeal circulation; and fourthly, it was necessary to develop biocompatible dialyzing membranes. Most of the essential knowledge was acquired by the end of the 19th century. Hemodialysis as a practical means of replacing kidney function started and developed in the 20th century. The original hemodialyzers, using celloidin as a dialyzing membrane and hirudin as an anticoagulant, were used in animal experiments at the beginning of the 20th century, and then there were a few attempts in humans in the 1920s. Rapid progress started with the application of cellophane membranes and heparin as an anticoagulant in the late 1930s and 1940s. The explosion of new dialyzer designs continued in the 1950s and 1960s and ended with the development of capillary dialyzers. Cellophane was replaced by other dialyzing membranes in the 1960s, 1970s, and 1980s. Dialysis solution was originally prepared in the tank from water, electrolytes, and glucose. This solution was recirculated through the dialyzer and back to the tank. In the 1960s, a method of single-pass dialysis solution preparation and delivery system was designed. A large quantity of dialysis solution was used for a single dialysis. Sorbent systems, using a small volume of regenerated dialysis solution, were developed in the mid 1960s, and continue to be used for home hemodialysis and acute renal failure. At the end of the 20th century, a new closed system, which prepared and delivered ultrapure dialysis solution preparation, was developed. This system also had automatic reuse of lines and dialyzers and prepared the machine for the next dialysis. This was specifically designed for quotidian home hemodialysis. Another system for frequent home hemodialysis or acute renal failure was developed at the turn of the 21st century. This system used premanufactured dialysis solution, delivered to the home or dialysis unit, as is done for peritoneal dialysis. PMID- 18394052 TI - Sustained low-efficiency dialysis with filtration (SLEDD-f) in the management of acute sodium valproate intoxication. AB - Hemodialysis is only infrequently used in drug overdosage situations. The efficacy of hemodialysis to remove the drug depends upon the pharmacokinetics and pharmacodynamics of the drug. At normal therapeutic concentrations, valproic acid is predominantly protein bound and therefore removal by hemodialysis is limited. In an overdose situation, protein binding is rapidly saturated and therefore the substantially larger quantities of the free drug can rapidly cause toxicity. Slow low-efficient daily diafiltration (SLEDD) has not previously been utilized in a drug overdose situation. We report the effective use of SLEDD to remove high toxic concentrations of valproic acid in an overdose situation. Slow low efficient daily diafiltration also prevented the rebound phenomenon that can occur as the excess drug is released from its protein-bound stores. Hybrid dialysis therapies deserve further evaluation in the management of other poisonings where extra-corporeal therapy is indicated. PMID- 18394053 TI - Prescribing and monitoring hemodialysis in a 3-4 x/week setting. AB - The basics of targeting, writing, adjusting, and monitoring a hemodialysis prescription are reviewed for patients being dialyzed 3 or 4 times a week. K/DOQI 2006 adequacy guidelines and practice recommendations are reviewed, and a practical method using a variety of nomograms is suggested to monitor and adjust the desired level of Kt/V. PMID- 18394054 TI - Glucose in the dialysate: historical perspective and possible implications? AB - Hemodialysate solutions often contain high concentrations of glucose (up to 200 mg/dL). The historical reasons for the addition of glucose to the dialysate included: (1) aid in performance of ultrafiltration and (2) minimization of nutritional (caloric) losses during dialysis. However, recent experimental evidence supports the fact that exposure to high levels of glucose may be pro inflammatory. Given the high morbidity and mortality associated with dialysis and its linkage to chronic inflammation, the routine use of glucose in the dialysate may warrant reexamination. This review examines the utility of glucose in the dialysate and discusses the potential implications on chronic inflammation in patients with end-stage renal disease. While there is currently no evidence for a casual relationship between dialysate glucose concentration and the chronic inflammation seen in ESRD, this possibility is explored. PMID- 18394055 TI - Endophthalmitis: a rare complication of arteriovenous fistula infection. AB - Vascular access infection is a frequent problem in patients undergoing maintenance hemodialysis. Infection of arteriovenous fistula (AVF) is less common than dialysis catheter-associated infection. Previous case reports described endophthalmitis secondary to hemodialysis catheter-related infection, but not secondary to native AVF infection. We report a rare patient of endophthalmitis as a metastatic infection of AVF cannulation site abscess. A 19-year-old girl on maintenance hemodialysis for the past 2 years has presented with a history of fever, chills, and rigor of 3-days duration and painful dimness of vision in the left eye of 1-night duration. It was followed by redness of the eye, photophobia, and ocular discharge. On examination, the patient was febrile with an abscess near cannulation site of AVF. There was no perception of light in the left eye, conjunctiva was congested, cornea was clear, hypopyon present, and pupil was mid dilated, not reacting to light. Lens was clear. Vitreitis and exudative retinal detachment was present. Methicillin sensitive Staphylococcus aureus was isolated from blood, pus from AVF abscess and vitreous fluid. Diagnosis of endophthalmitis was confirmed by B-scan ultrasound. She was treated with both intravenous and intraocular antibiotics and drainage of pus from AVF abscess and therapeutic vitrectomy. Though arteriovenous abscess responded to sensitive antibiotics and drainage, vision has not improved much. Strict aseptic precautions during regular AVF cannulation are required. Lapses may lead to loss of vision apart from described complications like access closure, endocarditis, and osteomyelitis. PMID- 18394056 TI - Gustatory sweating recurring on peritoneal dialysis but resolving during periods of hemodialysis. AB - Gustatory sweating is a rare disorder characterized by profuse sweating on the forehead, face, scalp, and neck occurring soon after ingesting food, which has been reported in diabetic patients. The mechanism is thought to be triggered by taste buds and not gastric stimulation. We report a case where gustatory sweating repeatedly developed on peritoneal dialysis that resolved on periods of hemodialysis. A 32-year-old woman with diabetic end-stage renal disease developed gustatory sweating shortly after beginning continuous ambulatory peritoneal dialysis despite excellent clearances. After 5 months, she changed to hemodialysis for 2 months and noticed complete resolution of her gustatory sweating; however, after her return to peritoneal dialysis 2 months later, her gustatory sweating recurred. While on peritoneal dialysis, she was treated with clonidine, which resulted in improvement but not resolution of her symptoms as had occurred on hemodialysis. Another period on hemodialysis resulted in the resolution of her symptoms that returned again after restarting peritoneal dialysis. Clonidine provided incomplete relief while topical glycopyrrolate was effective and without complications. We report recurrent gustatory sweating on peritoneal dialysis that resolved with hemodialysis. We have no data to suggest that intra-abdominal stimulation played a role, but rather that despite excellent clearances neuropathy may have played a role. Treatment with topical glycopyrrolate may be safe and effective given every third day if clonidine is ineffective. PMID- 18394057 TI - A king-size dialysis bed: a couple on hemodialysis for more than 30 years. AB - Dialysis patients exhibit a higher morbidity and mortality rate than those in the general population of comparable age. Survival on dialysis has become significantly longer and is mainly related to comorbid factors. Patients are usually the main research subjects, but caregivers play a pivotal role in patients' well-being. Here we describe the remarkable case of wife and husband both on hemodialysis for 32 years. PMID- 18394058 TI - Determinants of C-reactive protein in chronic hemodialysis patients: relevance of dialysis catheter utilization. AB - Biomarkers of inflammation, especially C-reactive protein (CRP), have been consistently shown to predict poor outcomes in chronic hemodialysis (CHD) patients. However, the determinants of CRP and the value of its monitoring in CHD patients have not been well defined. We conducted a retrospective cohort study to evaluate possible determinants of the inflammatory response in CHD patients with a focus on dialysis catheter utilization. Monthly CRP were measured in 128 prevalent CHD patients (mean age 56.6 years [range 19-90], 68% African Americans, 39% diabetics [DM]) over a mean follow-up of 12 months (range 2-26 months). There were a total of 2405 CRP measurements (median 5.7 mg/L; interquartile range [IQR] 2.4-16.6 mg/L). The presence of a dialysis catheter (p<0.002), cardiovascular disease (p=0.01), male gender (p=0.005), higher white blood cell count (p<0.0001), elevated phosphorus (p=0.03), and lower cholesterol (p=0.02) and albumin (p<0.0001) concentrations were independent predictors of elevated CRP in the multivariate analysis. Additionally, CRP levels were significantly associated with the presence of a catheter, when comparing the levels before and after catheter insertion (p=0.002) as well as before and after catheter removal (p=0.009). Our results indicate that the presence of a hemodialysis catheter is an independent determinant of an exaggerated inflammatory response in CHD patients representing a potentially modifiable risk factor. PMID- 18394059 TI - Prediction of hemodialysis sorbent cartridge urea nitrogen capacity and sodium release from in vitro tests. AB - In sorbent-based hemodialysis, factors limiting a treatment session are urea conversion capacity and sodium release from the cartridge. In vitro experiments were performed to model typical treatment scenarios using various dialyzers and 4 types of SORB sorbent cartridges. The experiments were continued to the point of column saturation with ammonium. The urea nitrogen removed and amount of sodium released in each trial were analyzed in a multi-variable regression against several variables: amount of zirconium phosphate (ZrP), dialysate flow rate (DFR), simulated blood flow rate (BFR), simulated patient whole-body fluid volume (V), initial simulated patient urea concentration (BUNi), dialyzer area permeability (KoA) product, initial dialysate sodium and bicarbonate (HCO3i) concentrations, initial simulated patient sodium (Nai), pH of ZrP, creatinine, breakthrough time, and average urea nitrogen concentration in dialysate. The urea nitrogen capacity (UNC) of various new SORB columns is positively related to ZrP, BFR, V, BUNi, and ZrP pH and negatively to DFR with an R2 adjusted=0.990. Two models are described for sodium release. The first model is related positively to DFR and V and negatively to ZrP, KoA product, and dialysate HCO3i with an R2 adjusted=0.584. The second model incorporates knowledge of initial simulated patient sodium (negative relationship) and urea levels (negative relationship) in addition to the parameters in the first model with an R2 adjusted=0.786. These mathematical models should allow for prediction of patient sodium profiles and the time of column urea saturation based on simple inputs relating to patient chemistries and the dialysis treatment. PMID- 18394060 TI - Impact of intradialytic exercise on arterial compliance and B-type natriuretic peptide levels in hemodialysis patients. AB - Cardiovascular (CV) disease is the most common cause of mortality in end-stage kidney disease (ESKD), and arterial stiffness, measured by pulse wave velocity (PWV), is an independent predictor of all-cause and CV mortality. B-type natriuretic peptide (BNP) levels are high in patients with CV disease and ESKD, and increases in BNP may also be a marker of CV risk. Regular exercise has many benefits on quality of life and physical strength and may also improve CV risk, but few studies have addressed the impact of exercise on CV risk in ESKD. We performed a prospective cross-over trial in 19 hemodialysis (HD) patients to assess the impact of regular exercise on surrogate markers of CV risk-arterial compliance and BNP levels. Exercise involved the use of a bicycle ergometer for minimum 30 min at each HD session for 3 months, with a 1-month washout period. Group A (n=9) exercised for the first 3 months only, while group B (n=10) performed no intradialytic exercise initially and exercised for 3 months at cross over (month 4). Pulse wave velocity was performed using a SphygmoCor device, with concurrent measurements of BNP and other serum markers, at the commencement of the study, at 3 months, and on completion. The mean PWV (A: 10.4+/-3.1 m/s, B: 9.8+/-3.8 at baseline) showed a trend toward improvement with exercise (A: 8.7+/ 2.7, p=0.07), and no significant change without (B: 10.5+/-3.6, p=0.31). After cross-over, there was an increase in PWV in group A with cessation of exercise (9.75+/-2.4, p=0.01 vs. 3 months) and an improvement in group B with exercise (9.33+/-2.3, p=0.11 vs. 3 months). When comparing PWV after 3 months of exercise vs. 3 months of no exercise (paired t test), there was a significant difference in favor of exercise (9.04+/-0.59 vs. 10.16+/-0.74, p=0.008). The mean BNP levels following 3 months of exercise were also lower than those after 3 months of no exercise (504.4+/-101.2 vs. 809.4+/-196.1[N<100], p=0.047). There was an overall improvement in PWV, and to a lesser extent BNP levels, with 3 months of intradialytic exercise compared with no exercise, suggesting that regular exercise in ESKD may be associated with improvements in arterial compliance and a reduction in CV risk. PMID- 18394061 TI - A new safety device for hemodialysis. AB - Accidental venous needle dislodgement during hemodialysis may cause serious bleeding including a sometimes fatal outcome. The venous pressure gauge of the dialysis monitor does not react when dislodgement occurs. A sensor patch put as an adhesive over the venous needle puncture site connected to an alarm unit by an optic fiber has been clinically tested in 5 dialysis departments. A small amount of blood on the sensor activates a light and sound alarm. A simple questionnaire was filled out by the nurses at each dialysis concerning their feeling of safety when the new device was used. Forty-one patients, mean age 65 years, have tested the new safety device. Two hundred test dialyses were studied, after exclusion of 13 tests. One hundred seventy-nine tests reacted positively on blood. In another 6 dialyses, a warning light appeared on the alarm unit indicating a failure in the sensor patch. Thus, the alarm functioned in 92.5% of all tests. After a small modification of the sensor patches there were only 2 dialyses (2/71) without an activated alarm on blood, i.e., 97.2% positive alarm reactions. The answers of the nurses indicated that they had an increased feeling of safety when using the new safety device, with a mean value of 3.4 points on a visual scale from 0 to 5 where 5 meant very much increased safety. In a situation when the dialysis monitors today do not react on bleedings from venous needle dislodgements, the new alarm safety device fulfils a known shortage in routine dialysis safety. In situations where supervision during a dialysis session may be insufficient as, for example, in home hemodialysis and self-care dialysis or in other situations when the patient is sleeping, the device may be life saving. PMID- 18394062 TI - Clinical correlates and treatment of bone/joint pain and difficulty with sexual arousal in patients on maintenance hemodialysis. AB - Bone/joint pain and difficulty with sexual arousal are prevalent, frequently severe, and potentially treatable in patients on maintenance hemodialysis. However, the mediators and adequacy of treatment for these symptoms have been less well studied. We sought to assess the clinical correlates and treatment of these symptoms in patients receiving chronic hemodialysis. Using the Dialysis Symptom Index, we assessed the presence and severity of bone/joint pain and difficulty with sexual arousal in 75 patients on chronic hemodialysis. Associations of demographic and clinical variables with these 2 symptoms were assessed. We also recorded the use of analgesics for bone/joint pain and, among men, phosphodiesterase-5 inhibitors for difficulty with sexual arousal. Twenty eight patients (37%) reported bone/joint pain, of whom 20 (71%) described it as moderate to severe. Nineteen of 50 male patients (38%) reported difficulty with sexual arousal, which was described as moderate to severe by 15 (79%). Patients' demographic and clinical characteristics were not correlated with bone/joint pain. Among men, there were no correlations between patients' demographic and clinical characteristics and difficulty with sexual arousal. Only 48% of patients with bone/joint pain were receiving analgesics, while 21% of men who described difficulty with sexual arousal were receiving phosphodiesterase-5 inhibitors. Demographic, clinical, and dialysis-related variables are poorly correlated with bone/joint pain and difficulty with sexual arousal. However, these symptoms are prevalent and under-treated, which should spur efforts to assess the impact of improving provider assessment and treatment of these symptoms on patient outcomes, including health-related quality of life. PMID- 18394063 TI - Statins are associated with a reduced risk of bone fracture in hemodialysis (HD) patients. AB - The Dialysis Outcomes and Practice Patterns Study reported a statistically non significant protective effect of HMG-co reductase inhibitors (statins) on bone fracture risk in hemodialysis (HD) patients. We sought to determine whether statin exposure was associated with reduced risk of bone fracture in our HD population. This was a retrospective cohort study of 174 prevalent HD patients. Fracture data are abstracted from the medical record. Subjects were considered to be on a statin if they were exposed at any time since the date of dialysis initiation. The subjects were 174 HD patients (68.4% male) with a median age of 69.1 and age range from 25.2 to 96.3 years. The median age at initiation of HD was 62.5, ranging from 15.2 to 90.5 years. The mean (SD) dialysis vintage was 7.3 (4.5) years. Seventy-seven subjects (44.3%) had statin exposure. There were a total of 54 first bone fractures. There was a positive correlation between bone fracture and dialysis vintage (p=0.023) and a negative association between bone fracture and statin exposure (p=0.044). Those with statin exposure had a higher prevalence of CAD (p=0.030) compared with those not exposed. Logistic regression analysis (stepwise, alpha=0.05) was performed with dependent variable bone fracture and independent variables age at HD initiation (forced), dialysis vintage, gender (forced), prednisone use (forced), and statin exposure. The significant predictors of bone fracture (R2=0.14, p=0.004) were age at HD initiation (p=0.016), dialysis vintage (p=0.007), and absence of statin exposure (p=0.019). Statin exposure appears to be associated with a reduced frequency of bone fracture in HD patients. Future studies evaluating the potential anabolic effect of statins on bone are required. PMID- 18394065 TI - Proximodistal alignment of the canine patella: radiographic evaluation and association with medial and lateral patellar luxation. AB - OBJECTIVES: To evaluate the contribution of proximodistal alignment of the patella to patellar luxation, and to evaluate the structures contributing to proximodistal alignment of the patella relative to the femoral trochlea. STUDY DESIGN: Retrospective study using a convenience sample. ANIMALS: Medium to giant breed dogs (n=106). METHODS: Medical records and stifle radiographs of 106 dogs were reviewed. Radiographic measurements evaluated the proximodistal alignment of the patella with respect to the femoral trochlea, distal aspect of the femur, and proximal aspect of the tibia. Measurements were compared between dogs with clinically normal stifles (controls; n=51 dogs, 66 stifles), and dogs with a clinical diagnosis of medial patellar luxation (MPL, n=46 dogs, 65 stifles) or lateral patellar luxation (LPL, n=9 dogs, 11 stifles) using ANOVA. RESULTS: In dogs with MPL, the ratio of patellar ligament length (PLL) to patellar length (PL) was increased, as was the ratio of the distance from the proximal aspect of the patella to the femoral condyle (A) to PL (P<.0001). Dogs with LPL had a decreased A:PL (P=.003) and an increased ratio of the proximal tibial length (PTL) to distal tibial width (DTW; P=.009). CONCLUSIONS: MPL is associated with a relatively long patellar ligament and patella alta in medium to giant breed dogs. LPL is associated with a relatively long proximal tibia and patella baja. Values for PLL:PL>2.06 and A:PL>2.03 are suggestive of the presence of patella alta, whereas a value for A:PL<1.92 is suggestive of patella baja. CLINICAL RELEVANCE: Measurements of both PLL:PL and A:PL are recommended in dogs with patellar luxation, and surgical correction should be considered in those with abnormal values. PMID- 18394066 TI - Comparison of different tube materials and use of Chinese finger trap or four friction suture technique for securing gastrostomy, jejunostomy, and thoracostomy tubes in dogs. AB - OBJECTIVE: To compare Chinese finger trap (CFT) and 4 friction suture (FFS) techniques to secure gastrostomy (GT), jejunostomy (JT), and thoracostomy (TT) tubes of different materials. STUDY DESIGN: Prospective experimental study. ANIMALS: Canine cadavers (n=20). METHODS: Randomly, GT (n=20), JT (20), and TT (20) were inserted using 2 different suture techniques (10 for each tube type) and either silicone or another material (10 for each type). Axial distraction was applied to each tube until failure. Force and displacement to failure and failure mode were recorded and compared between techniques and materials for GT, JT, and TT. RESULTS: CFT failed most commonly by suture breakage whereas FFS failed mainly by tube slippage (P=.003). For GT, failure occurred more commonly by tube slippage (n=15; P<.001) whereas tube breakage was more common for JT (n=10; P<.001) and suture breakage for TT (n=14; P=.022). Silicone had higher force to failure than latex with GT, lower force to failure than red rubber with JT, and lower displacement to failure than polyvinylchloride with TT. CONCLUSIONS: Different failure modes occurred for CFT (suture breakage) and FFS (tube slippage) and among different tube types (tube slippage with GT, tube breakage with JT, and suture breakage with TT). Based on study results, CFT is preferred to FFS for anchoring silicone GT and TT. Silicone GT, red rubber JT, and polyvinylchloride TT were more secure than latex GT, silicone JT, and silicone TT, respectively. CLINICAL RELEVANCE: CFT should be preferred over FFS based on force and displacement to failure, but tube type and tissue reaction could influence anchoring strength. PMID- 18394067 TI - Endoscopic removal of a bronchial carcinoma in a dog using one-lung ventilation. AB - OBJECTIVE: To describe anesthetic management of endoscopic electrosurgical removal of a bronchial carcinoma, partially blocking the right main stem bronchus in a Cocker Spaniel. STUDY DESIGN: Clinical case report. ANIMALS: Dog with a bronchial carcinoma. METHODS: To allow sufficient space for the endoscope and to avoid an oxygen-rich gas mixture in the trachea, which carries the risk of an airway fire when electrocautery is used, a 1 lumen endobronchial tube (EBT) was inserted into the left main stem bronchus. One-lung ventilation (OLV) started with a volume-controlled ventilator was switched to pressure-controlled ventilation in combination with positive end-expiratory pressure (PEEP). RESULTS: Resection of the bronchial carcinoma was successful. The dog was hypercapnic throughout the procedure and a high alveolar-arterial oxygen gradient was measured. CONCLUSION: An EBT may be a feasible and safe option to provide OLV for bronchoscopic electrocautery with a closed thoracic cavity in dogs. CLINICAL RELEVANCE: EBT intubation for OLV should be considered as part of the anesthetic management of airway diseases treated with bronchoscopic electrocautery. PMID- 18394068 TI - Computed tomographic features of incomplete ossification of the canine humeral condyle. AB - OBJECTIVES: To describe computed tomographic (CT) features of canine elbows with incomplete ossification of the humeral condyle (IOHC) and investigate co-existing incongruence in the elbow joint. STUDY DESIGN: Case control study. ANIMALS: Dogs with IOHC (n=20; 38 elbows) and 25 normal elbows. METHODS: Elbows with IOHC and normal elbows were assessed by CT. Standardized dorsal and sagittal reconstructions were created at 3 levels using image analysis software to obtain single measurements of the humero-radial and humero-ulnar joint spaces. On dorsal plane reconstructions, joint space measurements were obtained at the center point of the humero-radial and humero-ulnar articulations. Joint incongruity was defined as the difference between the humero-radial and the humero-ulnar joint spaces. RESULTS: Nineteen dogs (95%), all Spaniel breeds, had either bilateral IOHC demonstrable as a saw-toothed intercondylar complete or incomplete hypoattenuating defect with hyperattenuating margins, or IOHC with contralateral humeral condylar fracture (HCF). Joint incongruity values for IOHC were compared with those of normal elbows. Significant differences were noted at the levels of the medial coronoid apex (P<.0001) and base (P<.004) indicative of humero-ulnar incongruence. Evidence of medial coronoid disease in 10 elbows (26%) and degenerative joint disease in 30 elbows (79%) was also found. CONCLUSIONS: Presence of elbow incongruence may be an underlying factor in failure of ossification centers to fuse leading to IOHC. CLINICAL RELEVANCE: IOHC is clearly defined by CT, and it should be considered in larger Spaniel breeds, with a chronic forelimb lameness or HCF. PMID- 18394070 TI - Relationship between objective and subjective assessment of limb function in normal dogs with an experimentally induced lameness. AB - OBJECTIVE: To evaluate the relationship between previously used subjective and objective measures of limb function in normal dogs that had an induced lameness. STUDY DESIGN: Prospective, blinded, and induced animal model trial. ANIMALS: Normal, adult, and mixed-breed dogs (n=24) weighing 25-35 kg. METHODS: Force platform gait analysis was collected in all dogs before and after induction of lameness. All gait trials were videotaped; 60 video trials were evaluated by 3 surgeons with practice limited to small animal orthopedics and 3 first year veterinary students in an effort to establish the relationship between subjective and objective measures of lameness. Evaluators were unaware of the force platform data. RESULTS: Concordance coefficients were low for all observers and were similar between students and surgeons. These values were further decreased when normal and non-weight bearing trials were removed. Agreement with the force platform data was low even when observers only had to be within +/-10% of the ground reaction forces. When repeat trials were evaluated surgeons had a much higher repeatability compared with students. CONCLUSIONS: Subjective evaluation of the lameness in this study varied greatly between observers and agreed poorly with objective measures of limb function. CLINICAL RELEVANCE: Subjective evaluation of gait should be interpreted cautiously as an outcome measure whether performed from a single or from multiple observers. PMID- 18394069 TI - Evaluation of a thoracoscopic technique using ligating loops to obtain large lung biopsies in standing healthy and heaves-affected horses. AB - OBJECTIVE: To evaluate use of pre-tied ligating loop to perform thoracoscopic, large lung biopsy in normal and heaves-affected horses. STUDY DESIGN: Prospective clinical study. ANIMALS: Normal (n=5) and heaves-affected (n=6) horses. METHODS: Lung biopsies, 1 from each hemithorax, were collected thoracoscopically using a pre-tied ligating loop. Horses were either normal (C) or heaves-affected with the latter being in remission (Ha) for the initial biopsy and in exacerbation (Hs) for the 2nd biopsy. Clinical variables, PaO(2), and PaCO(2) were used to determine the effect of surgical biopsy. Postoperative pneumothorax was monitored by serial thoracic radiographic examinations. RESULTS: Thoracoscopic lung biopsy (n=29, 22 procedures) was well tolerated by all horses. Complication rate was 31%, including 8 ligature slippage and 1 pulmonary hemorrhage. Intranasal oxygen was administered intraoperatively to 6 horses (2 C, 1 Ha, 3 Hs) with severe hypoxemia or labored breathing. There was a significant decrease in PaO(2) during surgery in horses not supplemented with oxygen. Postoperative pneumothorax (21/22 procedures) detected radiographically resolved within 3 weeks. CONCLUSION: Thoracoscopic lung biopsy using pre-tied ligating loops was minimally invasive, relatively inexpensive, and fairly efficient. Heaves-affected horses tolerated the surgery well, even when in exacerbation; however, the technique was associated with non life-threatening complications in 31% of the biopsies, most of which required correction with additional ligating loops or more sophisticated instrumentation. CLINICAL RELEVANCE: Using laparoscopic pre-tied ligating loop for thoracoscopically-assisted lung biopsy can be considered in horses with normal and impaired lung function but alternative instrumentation and access to intranasal oxygen must be available to the surgeon in case of complications. PMID- 18394071 TI - Gingival overgrowth in dogs associated with clinically relevant cyclosporine blood levels: observations in a canine renal transplantation model. AB - OBJECTIVE: To investigate the development of gingival hyperplasia in dogs after renal transplantation and administration of microemulsified cyclosporine A (MCsA). STUDY DESIGN: Experimental study. ANIMALS: Healthy adult mongrel dogs (n=5). METHODS: As part of study on renal transplantation, dogs administered MCsA (20 mg/kg/day), azathioprine, and prednisolone to prevent graft rejection were monitored for development of gingival changes. Prednisolone was discontinued after 3 months. MCsA dose was adjusted to maintain whole blood trough concentration of 400-700 ng/mL. Gingival change was evaluated by weekly examination and photodocumentation, and gingival biopsy for histopathology was performed at 28 weeks. RESULTS: One dog was lost because of acute graft rejection. Gingival hyperplasia developed in 3 of 4 dogs. The earliest gingival changes occurred in the interdental papillae at 20 weeks after transplantation. On histopathology, the underlying connective tissue was thickened and contained increase numbers of fibroblasts and inflammatory infiltrates. CONCLUSIONS: Long term immunosuppression with an MCsA-based treatment likely induces substantial gingival hyperplasia when therapeutic, immunosuppressive blood levels of MCsA were maintained for 32 weeks. CLINICAL RELEVANCE: MCsA is used for immune mediated diseases and preventing rejection after transplant in dogs. MCsA blood levels, and gingival hyperplasia should be monitored by routine examination of the interdental papilla in dogs administered MCsA for long periods. PMID- 18394072 TI - Sciatic neurapraxia as a complication in 1000 consecutive canine total hip replacements. AB - OBJECTIVES: To report the frequency of sciatic neurapraxia (SN) associated with total hip replacement (THR), to determine outcome (recovery rate) after SN associated with THR, and to identify potential causes of THR-associated SN in dogs. STUDY DESIGN: Prospective study. ANIMALS: Dogs (n=786; 1000 hips) that had THR. METHODS: Logistic regression was used to determine the association with post THR SN of the variables age, sex, breed, weight, body condition score, severity of presurgical pain, side (right, left), type of prosthesis fixation (cemented, cementless), duration of surgery, surgeon experience (chronologic order), traumatic presurgical luxation, and primary versus revision arthroplasty. RESULTS: The frequency of SN after THR was 19/1000 (1.9%). Two explanatory variables, age at surgery and duration of surgery, were significantly (P<.05) associated with increased probability of SN. Body weight (P=.09), traumatic presurgical luxation (P=.11), and revision versus primary surgery (P=.11) were marginally associated with increased probability of SN. All dogs with SN recovered fully. CONCLUSIONS: SN after THR is not uncommon and complete recovery usually occurs although the recovery time is highly variable. CLINICAL RELEVANCE: Although SN associated with THR typically resolves, surgeons should avoid iatrogenic sciatic nerve injury during THR. PMID- 18394073 TI - Clinical evaluation of the locking compression plate for fetlock arthrodesis in six thoroughbred racehorses. AB - OBJECTIVE: To report use of a locking compression plate (LCP) for fetlock arthrodesis in Thoroughbred racehorses after catastrophic disruption of the suspensory apparatus. STUDY DESIGN: Retrospective case study. ANIMALS: Racing Thoroughbreds (n=6) with a catastrophic breakdown injury of the suspensory apparatus. METHODS: Medical records (2004-2006) of horses that had fetlock arthrodesis using an LCP were reviewed. Preoperative recorded variables were: age, gender, affected limb, injury type and occurrence, limb support used and time to surgery. Recorded surgical variables were: implants used and cost, surgery time, and anesthetic recovery method. Postoperative recorded variables included: treatment, physical status, complications, and outcome. RESULTS: Of 6 Thoroughbred racehorses that had LCP arthrodesis, 4 were sound for breeding purposes the next year and 2 horses were euthanatized (1 at 16 days, 1 at 68 days) because of proximal interphalangeal joint (PIJ) luxation. No surgical complications were associated with implant application. Postoperative complications were similar to those reported for other arthrodesis techniques. CONCLUSIONS: LCP is a viable option for fetlock arthrodesis in Thoroughbred racehorses after catastrophic suspensory apparatus disruption. CLINICAL RELEVANCE: Adequate fetlock stability can be achieved with an LCP, and although the repair is likely more stable it is approximately 3 times more expensive than using a limited contact dynamic compression plate (LC-DCP) with cortical screws. PMID- 18394074 TI - Canine stifle positive contrast computed tomography arthrography for assessment of caudal horn meniscal injury: a cadaver study. AB - OBJECTIVE: To investigate the use of computed tomography (CT) arthrography in cadaveric canine stifles with particular emphasis on the diagnosis of meniscal injury. STUDY DESIGN: Prospective cadaver study. SAMPLE POPULATION: Pelvic limbs from adult Beagles (n=10). METHODS: After survey CT scan of each stifle oriented in the dorsal plane, positive contrast stifle CT arthrogram (CTA) was performed using the same slice orientation. Each stifle was then randomly allocated into 1 of 2 treatment groups: group A--arthrotomy, cranial cruciate ligament (CCL) transection and simulated injury to the caudal horn of the medial meniscus; group B--arthrotomy and CCL transection only. CT scan was repeated as before and post arthrotomy images were interpreted by a radiologist unaware of treatment grouping. RESULTS: The cranial and caudal cruciate ligaments, medial and lateral menisci, menisco-femoral ligament, and long digital extensor tendon were all identifiable on CTA images. CTA was 90% sensitive and 100% specific for diagnosing simulated caudal horn meniscal injury. CONCLUSIONS: Stifle CTA enables identification of intra-articular structures within the stifle and is a reliable method for identifying simulated meniscal injuries in a cadaver model. CLINICAL RELEVANCE: CTA imaging of the canine stifle has potential clinical value for detection of meniscal injury. PMID- 18394075 TI - Trephination of the equine rostral maxillary sinus: efficacy and safety of two trephine sites. AB - OBJECTIVE: To examine 2 rostral maxillary sinus (RMS) trephine sites for safety and efficacy using skulls of horses of varying age. STUDY DESIGN: Descriptive study. ANIMALS: Cadaveric equine skulls (n=40). METHODS: Two RMS trephination sites (rostral, caudal) were made in each skull. Radiographic projections using markers at each site were used to determine if sites were within the RMS and directly overlying a cheek tooth. Sinusotomy in 14 skulls was used to determine correlation between radiographic and anatomic location of trephine sites, and of the rostrolateral aspect of the maxillary septum. Age-related risk of trephine site being directly over a cheek tooth was determined using logistic regression. RESULTS: Trephine site was within the RMS in 98% of skulls using the rostral portal and 68% using the caudal portal. The rostral site was over a cheek tooth in 18% of skulls compared with 10% using the caudal site. There was a significant negative relationship between the trephine site overlying a tooth and horse age for rostral (P=.02) and caudal (P=.03) sites. The radiographic appearance of the maxillary septum correlated to the rostrolateral portion of the septum in 12 of 14 skulls that had sinusotomy. CLINICAL RELEVANCE: The rostral RMS trephine portal is more reliable than the caudal site for entering the RMS. Horses .05) than those with the 4.5 LC-DCP fixation. CONCLUSION: 5.5-LC-DCP fixation was superior to 4.5-LC-DCP fixation in resisting the static overload forces under palmarodorsal 4-point bending. There was no significant difference between 5.5-LC-DCP fixation and 4.5 LC-DCP fixation in resisting static overload forces under torsion; however, the 5.5-LC-DCP offers significantly less stability (80% of that of the 4.5-LC-DCP) in cyclic fatigue testing. CLINICAL RELEVANCE: The results of this in vitro study may provide information to aid in the selection of a biological plate for long bone fracture repair in horses. PMID- 18394078 TI - The diagnostic yield of conventional radiographs and computed tomography in dogs and cats with maxillofacial trauma. AB - OBJECTIVE: To compare the diagnostic yield of conventional radiographs and computed tomography (CT) images of the skulls of dogs and cats with maxillofacial trauma (MFT). STUDY DESIGN: Prospective study. ANIMALS: Dogs (n=9) and 15 cats with MFT. METHODS: CT-scans and skull radiographs (4 standard projections) for each animal were evaluated using a semi-quantitative scoring system for the ability to identify 26 predefined, clinically relevant anatomic features (Part 1), and 27 predetermined potential traumatic injuries (Part 2). For Part 1, mean scores for each anatomic feature were recorded for every view and imaging modality. For Part 2, studies were evaluated for the frequency of cases where each predetermined traumatic injury was identified. RESULTS: Part 1: On radiographs it was easy to identify 17 of 26 anatomic features whereas 6 features were very difficult or impossible to identify on any view. All structures were considered easy or very easy to identify on CT. Scores for CT were lower than radiographs for evaluating dental occlusion and the integrity of the mandibular body. Part 2: CT scans demonstrated 1.6 times more maxillofacial injuries for dogs and 2.0 times more for cats than conventional radiographs. The average number of MFT injuries per animal by radiographs and CT-scan was 4.8 and 7.6 in dogs, and 3.8 and 7.7 in cats, respectively. CONCLUSION: CT is superior to conventional skull radiography for identification of anatomic structures and traumatic injuries in dogs and cats. Skull radiography is useful for visualizing the mandibular body and dental occlusion. CLINICAL RELEVANCE: CT allows for accurate assessment, diagnosis and treatment planning of MFT in dogs and cats. PMID- 18394079 TI - Radiographic measurement of canine tibial angles in the sagittal plane. AB - OBJECTIVE: To describe a method for radiographic measurement of the mechanical joint angles of the tibia in the sagittal plane and report reference ranges for a population of dogs with cranial cruciate ligament disease, and specifically for Labrador Retrievers. STUDY DESIGN: Retrospective evaluation of radiographs. ANIMALS: Canine tibia (n=150) of dogs evaluated for cranial cruciate ligament disease; 104 tibiae were from Labrador Retrievers, 46 tibiae were from other medium- to large-breed dogs. METHODS: Medial-lateral radiographs were used to establish anatomic landmarks and measure the mechanical cranial distal tibial angle (mCrDTA) and mechanical caudal proximal tibial angle (mCaPTA). RESULTS: For all tibiae mean mCrDTA was 81.6 degrees and mCaPTA, 63.6 degrees . For Labrador Retriever tibiae mean mCrDTA was 81.7 degrees and mCaPTA, 63.8 degrees . For non Labradors, means mCrDTA was 81.5 degrees and mCaPTA, 63 degrees . Labrador Retrievers were significantly younger than non-Labradors in our study population. No significant differences were noted between Labrador Retrievers and non Labradors for mCrDTA or mCaPTA and correlations between mCrDTA and mCaPTA were neither strong nor significant. CONCLUSION: A method was established for radiographic measurement of mechanical joint angles of the canine tibia in the sagittal plane. Reference ranges are reported for a population of Labrador Retrievers and a population of non-Labradors with cranial cruciate ligament disease. CLINICAL RELEVANCE: Measurement method and reference ranges provided for mechanical tibial angles may be used to aid in diagnosis, surgical planning, and postoperative critique for hindlimb angular deformities. PMID- 18394080 TI - Transpalpebral eye enucleation in 40 standing horses. AB - OBJECTIVE: To report a technique for eye enucleation in standing sedated horses and to report outcome in 40 horses. STUDY DESIGN: Retrospective study. ANIMALS: Horses (n=40) requiring eye enucleation. METHODS: The eye was enucleated using a transpalpebral technique in 40 horses restrained in stocks and sedated. Anesthesia of orbital structures was provided by local nerve blocks and infiltration of the surgical site with local anesthetic solution. RESULTS: Affected eyes were successfully enucleated with the horse standing. Short-term complications included moderate swelling (5 horses) and wound discharge (1). Long term complications were not observed. CONCLUSIONS: A diseased eye can be safely enucleated with a horse standing. CLINICAL RELEVANCE: Enucleating an equine eye in the standing position eliminates the risks and costs of general anesthesia. PMID- 18394081 TI - Dermatologic Therapy issue on "New and emerging treatments in dermatology". PMID- 18394082 TI - New and emerging treatments in dermatology: acne. AB - Topical retinoids, benzoyl peroxide, azelaic acid, and topical and oral antibiotics remain the milestone of treatment for mild to moderate acne vulgaris. Oral isotretinoin is useful for the treatment of severe nodular acne, treatment resistant acne, and acne with a risk of physical or psychological scarring. Hormonal treatment in female acne is useful in resistant or late-onset acne. With increasing concerns regarding teratogenicity of isotretinoin and increasing antibiotic resistance, there is a clear need for therapeutic alternatives to these long-used treatments. Research in the pathogenesis of acne has allowed for new therapies and future perspectives regarding acne to evolve. They include low dose long-term isotretinoin regimens, insulin-sensitizing agents, 5alpha reductase type 1 inhibitors, topical photodynamic therapy, new combination formulations, dietary interventions, and antiinflammatory agents such as lipoxygenase inhibitors. PMID- 18394083 TI - Atopic dermatitis. AB - Atopic dermatitis (AD) is a common dermatologic disease that can occur at different ages with different clinical manifestations. The etiologic factors and the pathogenetic pathways of AD have been thoroughly investigated but as often happens, something more has to be yet elucidated before the statement, that we possess the overall comprehension of the disease, could be considered the truth. Treatment of AD is often challenging, because of the chronic course of the disease and the fact that even the best of therapies is affected by adverse events, intolerances, or the possible occurrence of contraindications resulting from a sudden change in the overall health status of the patient (e.g., the onset of some comorbidities). Hence, the need to know "how, when, with what, and why" to treat AD patients with the many therapeutic modalities is now in the hands of the dermatologists. PMID- 18394084 TI - Lyme borreliosis treatment. AB - Lyme borreliosis is the most common human tick-borne illness in the Northern Hemisphere. The causative agent is the spirochete Borrelia burgdorferi species complex, and the hard-shell ticks of the genus Ixodes is responsible for pathogen transmission from animals to humans. The incidence of the disease is increasing year by year and although lyme disease is not fatal, it can affect the skin, heart, nervous, and musculoskeletal system with an impairment of quality of life. The appropriate diagnosis of lyme disease should be promptly treated by antibiotics to prevent late stage of the disease. The choice of antibiotics depends on many factors such as the stage of the disease, the drug efficacy, adverse effects, type of delivery, duration of treatment, and cost. Treatment failure occurs as a result of many reasons, re-infection is possible. The recommended treatment schedule in the Czech Republic is presented. PMID- 18394085 TI - Vitiligo: new and emerging treatments. AB - Vitiligo is a cosmetically disfiguring condition, and, although there is no therapeutic full solution yet, some treatment may induce good results in most patients. The disease can be successfully treated with various medical options. Both nonfocused or focused narrowband ultraviolet B phototherapy represents the current treatment of choice, to minimize side effects and reach optimal clinical results. Topical novel approaches are also considered. Surgical methods, consisting of autologous transplantation methods, is generally recommended for focal/stable vitiligo, after medical therapy has failed. Finally, for patients with extensive vitiligo, depigmentation of the residual melanin should be taken into account. PMID- 18394087 TI - From conventional to cutting edge: the new era of biologics in treatment of psoriasis. AB - Psoriasis is a chronic T-cell-mediated inflammatory disease of the skin and joints that affects 1-3% of the world population. Conventional treatments for moderate to severe psoriasis are associated with broadband immunosuppression and/or organ toxicities that can be problematic when used long term. Advances in the understanding of psoriasis pathogenesis have led to targeted therapy in the form of biologics. These agents have gained popularity as safe, effective, and convenient alternatives for the treatment of chronic, moderate to severe plaque psoriasis. This review will focus on the five main biologics used in the treatment of moderate to severe plaque psoriasis: efalizumab, alefacept, etanercept, infliximab and adalimumab. Mechanisms of action, guidelines for usage, efficacy data, and safety concerns will be discussed for each biologic. In addition, the new Th17 biologics and their role in psoriasis pathogenesis will also be examined. PMID- 18394086 TI - Esthetic and cosmetic dermatology. AB - The field of esthetic and cosmetic dermatology has gained remarkable interest all over the world. The major advantage of recent years is the high scientific levels of the most significant new developments in techniques and pharmacotherapy and other nonsurgical approaches. The present paper reviews selected fields of interest under this view. Sexual hormones are involved in the aging process of men and women. Skin function, in particular the epidermal barrier, is affected by a loss of endocrine activity. Hormone replacement therapy has only recently been introduced in treatment of aging males. This is an area of gender-medicine in dermatology with a strong well-aging attempt. Botulinum toxin therapy for hyperfunctional lines has become not only well-established but evidence-based medicine on its highest level. Recent advantages were gained in objective evaluation and monitoring the effect. Digital imaging techniques with various facets have been introduced to assess the achievements of treatment in the most objective way. This may become an example for other techniques as peeling, laser therapy, or radiofrequency in esthetic and cosmetic dermatology. Botulinum toxin has become a valuable tool for brow lifts. Details of the technique are discussed. Cellulite is a strongly female gender-related condition. During the past decades numerous treatments had been recommended but only recently a more critical scientific approach led to improvements in therapy of this common and disfiguring condition. Three major approaches are developed: (a) skin loosing with techniques such as subcision, (b) skin tightening with radiofrequency and other approaches, and (c) improving circulation in blood and lymphatic microvasculature using both physical treatments and pharmacotherapy. The last two chapters are devoted to body sculpturing by lipotransfer and lipolysis. Lipotransfer for facial or body sculpturing has a history of about 100 years. Nevertheless, recently the role of adult stem cells in adipose tissue has gained much interest. By optimizing the harvesting, storage, and transplantation of adipose tissue, remarkable long-standing results have been obtained. Here the present authors will focus on midface contouring, where lipotransfer competes with dermal fillers. Lipolysis is another effective tool in body sculpturing. The present authors will focus on recent advances in laser-assisted lipolysis for delicate body sculpturing in the submental region but also for gynecomastia abdominal region, flanks, and hips. In conclusion, esthetic and cosmetic dermatology has become a scientific-based subspeciality of dermatology with evidence-based treatments and a great variety of high-tech approaches to provide more effective, more selective, and safer therapeutic options. PMID- 18394088 TI - What's new in dermatologic therapy. AB - Dermatology is constantly evolving, with ever expanding knowledge of the diagnosis, pathogenesis, and treatment of skin disease. This article reviews important recent advances in dermatologic therapy and their impact on patient management. PMID- 18394091 TI - Case 1: an infant with an extracranial protuberance (case presentation). PMID- 18394092 TI - Case 2: painful red plagues in a 6-year-old child (case presentation). PMID- 18394093 TI - Radiation, retardation and the developing brain: time is the crucial variable. AB - BACKGROUND: Widespread radiation is a threat unique to the modern world. A recent report reveals that sub-clinical damage to human foetuses between 8 and 25 weeks of gestation can result in cognitive deficits still manifest 16-18 years after birth. These previously unrecognised, long-term effects are apparently produced by a relatively short pulse of exposure to radioactive fallout at levels that were previously thought not to be deleterious. This idea is plausible given the nature of the developmental events occurring in the brain during this period of gestation. CONCLUSION: This exposed population should be examined for other neurological and psychiatric syndromes. If these findings are corroborated, in the event of future radiation exposures, steps should be taken to shield pregnant women who are within this window of vulnerability. PMID- 18394094 TI - Tick-borne encephalitis (TBE) in childhood. PMID- 18394095 TI - Tick-borne encephalitis in Styrian children from 1981 to 2005: a retrospective study and a review of the literature. AB - BACKGROUND: Tick-borne encephalitis in children appears to be more benign than in adults and shows also a more favourable outcome. Only some authors report of sequelae like paralysis, paresis or seizures and behavioural abnormalities. The aim was to describe the clinical features of tick-borne encephalitis in children with special attention to sequelae and to review the literature. METHODS: Retrospective review of all charts of children with serologically confirmed tick borne encephalitis hospitalised in Styria between 1981 and 2005. RESULTS: One hundred sixteen children were diagnosed with tick-borne encephalitis. Ninety-two children (79.3%) developed meningitis and 24 (20.7%) meningoencephalitis. Eleven patients with meningoencephalitis showed somnolence, 5 confusion, 5 tremors, 2 facial palsy, 1 ataxia, 1 epileptic seizure and 1 hemi paresis. Seven patients had to be admitted to the intensive care unit. Two children (1.7%) developed long time neurological sequelae: one epileptic seizure with requirement of antiepileptic therapy and one left-sided hemi paresis. One hundred twelve children had been not and 3 incompletely vaccinated against tick-borne encephalitis. Only one child had been fully vaccinated according to the Austrian vaccination schedule. CONCLUSION: Our study provides further evidence that tick borne encephalitis in children has a substantial morbidity and in single cases severe long-time neurological sequelae are observed. PMID- 18394096 TI - Maternal use of loperamide in early pregnancy and delivery outcome. AB - AIM: To study delivery outcome including presence of infant congenital malformations after maternal use of loperamide in early pregnancy. METHODS: Using the Swedish Medical Birth Register, women reporting the use of loperamide in early pregnancy were compared with other women for the period July 1, 1995-2004. Odds ratios (OR) or risk ratios (RR) were estimated after adjustment for some variables, which were associated with the use of loperamide (year of delivery, maternal age, parity, smoking, number of previous miscarriages). RESULTS: Characteristics of women using loperamide in early pregnancy were identified. An analysis of concomitant drug use indicated that only few of the women had inflammatory bowel disease. The risk of any congenital malformation was increased (OR = 1.43, 95% CI 1.04-1.96), based on 43 cases, but no major contributing type could be identified. The risk of hypospadias was significantly increased (RR = 3.2, 95% CI 1.3-6.6), based on seven cases. A statistically significant increase was seen also for placenta previa, large for gestational age and caesarean section. CONCLUSION: Maternal use of loperamide in early pregnancy may be associated with a moderate risk increase for a malformation in the infant. The finding should be evaluated from other large studies. PMID- 18394097 TI - Assessment of foetal exposure to cigarette smoke after recent implementations of smoke-free policy in Italy. AB - AIM: We investigated foetal exposure to cigarette smoke following maternal active and passive smoking at the end of the pregnancy in a cohort of mother-infant dyads after recent implementation of the Italian smoke-free legislation and information campaigns against smoking. METHODS: Study subjects were 979 mothers and their newborns attended at one of the major Florence hospitals between the end of 2004 and during 2005, when smoking banning legislation in all the enclosed places of employment was completely enforced. A structured questionnaire was completed by mothers and cord serum cotinine was used as a biomarker of foetal exposure to maternal smoke at the end of the pregnancy. RESULTS: Women (53.9%) declared to be non-smokers not exposed to environmental tobacco smoke (ETS), 33.5% to be exposed non-smokers, while 12.6% reported daily smoking during pregnancy. Based on the levels of cord serum cotinine, 78.9% newborns were not exposed to ETS, 13.5% were exposed to ETS and 7.7% were exposed to active maternal smoke. CONCLUSIONS: Smoke-free policies can be an effective strategy for reducing both first-hand and second-hand exposure to cigarette smoke in pregnant women and their newborns. Due to the implementation of smoke-free legislation and information campaigns against smoking, a significant trend towards less active smoking during pregnancy was observed in our study cohort, as well as a minimal exposure to ETS in non-smoking pregnant women and their newborns. PMID- 18394098 TI - Short-term outcome predictors in infants born at 23-24 gestational weeks. AB - AIM: Outcome is uncertain in infants born at 23-24 gestational weeks. The aim of the present study was to identify possible early predictors of outcome in these infants. MATERIALS AND METHODS: Data from the Swedish medical birth register (MBR) for live-born infants with gestational ages (GAs) 23 and 24 weeks, born during the time-period 2000-2002, were analysed in relation to short-term outcomes, that is survival and survival without severe brain damage (intraventricular haemorrhage [IVH] grades 3 and 4 and/or periventricular leukomalacia [PVL]). RESULTS: In 57 infants born at 23 gestational weeks, survival was associated with birthweight (BW) (p = 0.018) and 5-min Apgar score (p = 0.020) on univariate analyses. In 99 infants born at 24 weeks of gestation, survival without severe brain damage correlated with BW (p = 0.039), birth type (singleton/multiple) (p = 0.017) and Apgar score at 1, 5 and 10 min (p = 0.028, 0.014 and 0.030, respectively). The best model for predicting survival without severe brain damage in infants born at 24 gestational weeks was based on 5-min Apgar score and birth type. The small number of live-born infants at 23 weeks of gestation did not allow for multiple logistic regression analyses. CONCLUSION: The 5-min Apgar score is associated with short-term outcome in live-born infants at 23-24 gestational weeks. The association is stronger for infants born at 24 weeks of gestation. PMID- 18394099 TI - Nasal respiratory support in premature infants: short-term physiological effects and comfort assessment. AB - AIM: To evaluate the effects of nasal respiratory support on physiologic parameters and comfort of premature infants, when compared to spontaneous breathing without nasal respiratory support. METHODS: This was a prospective, randomized, controlled, cross-over clinical study. Infants were enrolled into the study when in 'stable' condition (when discontinuation of nasal respiratory support was considered appropriate). Infants were randomized to receive first 3 h of nasal respiratory support (nasal continuous positive airway pressure or nasal intermittent mandatory ventilation) or to spontaneous breathing, and then were crossed-over to the other assignment. Each infant served as his own control. RESULTS: Fifty-four infants were included in the study (birth-weight: 1528 +/- 545 g; gestational age: 30.5 +/- 2.7 weeks). Average values of systolic, diastolic and mean blood pressure and discomfort score were significantly higher while respiratory rate was significantly slower on nasal respiratory support compared to spontaneous breathing. Heart rate was comparable on both modes. CONCLUSIONS: Nasal respiratory support in 'stable' premature infants is associated with increased blood pressure and increased discomfort, despite a decreased respiratory rate. The clinical importance of these effects is modest. Medical teams should consider these effects and balance its need with its adverse effects according to the clinical condition. PMID- 18394100 TI - Neurodevelopmental outcomes of very low-birth-weight infants after enteral glutamine supplementation in the neonatal period. AB - AIM: To determine the effect of neonatal glutamine-enriched enteral nutrition in very low birth weight (VLBW) infants on neurodevelopmental outcome at 2 years of age. METHODS: Eighty-eight out of one hundred two infants participating in the initial study were eligible for the follow-up study (13 died, one exclusion due to a chromosomal abnormality). Neurodevelopmental outcome (neurologic status, vision, hearing and Mental Developmental Index (MDI) and Psychomotor Developmental Index (PDI) of the Bayley Scales of Infant Development II) was evaluated at the corrected age of 2 years. To adjust for potential confounders, data were analyzed by multiple linear or logistic regression (for continuous and dichotomous variables, respectively) RESULTS: Seventy-two out of eighty-eight (82%) infants participated in the follow-up study: 40 and 32 infants in glutamine supplemented and control groups, respectively. The incidence of neither an MDI nor a PDI or=70 from a population of all the 107 children with hydrocephalus born in western Sweden in 1989-1993 were examined and compared with age- and gender-matched controls. The neuropsychological assessment of the school-aged child (NIMES) test battery was used. RESULTS: The children with hydrocephalus differed significantly from controls in all functions apart from registration skills and recognition. Learning, memory and executive functions were all impaired. Twenty children with infantile hydrocephalus did not differ from those with hydrocephalus associated with MMC. Also, children with an IQ of >84 performed significantly worse than controls. CONCLUSIONS: Despite an IQ of >or=70, children with hydrocephalus had significantly impaired learning, memory and executive functions. When major brain lesions resulting in learning disability had been excluded, the hydrocephalus, rather than the underlying aetiology, was most important for the development of cognitive functions. PMID- 18394106 TI - How well do care providers know the children with developmental disabilities they care for? AB - AIM: To assess the knowledge from memory of caregivers about the most significant impairments contributing to additional care needs in children with developmental disabilities in therapeutic toddler groups. METHODS: Children's needs for additional care due to impairments of physical health, motor, sensory, mental and voice and speech functions were separately classified using the capacity profile (CAP). Twenty-three therapists and teachers of toddler groups in two regional centres for paediatric rehabilitation assessed the CAP individually, unprepared and without consulting their notes or the clinical record. These CAP scores (150 CAPs of 44 children) were compared with those based on the clinical record using weighted kappa statistics. RESULTS: Weighted kappa values for the two sets of CAP scores ranged from 0.22-0.74 (median 0.53), with the lowest scores for the sensory domain (median 0.32, range 0.22-0.52) and the highest scores for the motor domain (median 0.62, range 0.56-0.74). CONCLUSION: Team members in general had only moderate remembered knowledge of the current impairments determining need of additional care. Remembered knowledge was the poorest for domains not easily observable, such as sensory functions. As this knowledge is essential for optimizing the child's daily environment, improvement of this type of knowledge should be facilitated. PMID- 18394107 TI - Maternal employment and breastfeeding: results from the longitudinal study of Australian children. AB - AIM: To investigate the effect of maternal postnatal employment on breastfeeding duration in Australia in the first 6 months after birth. METHOD: Secondary data analysis of the infant data (2004) from the Longitudinal Study of Australian Children (LSAC). Complete maternal and breastfeeding data were available for 3,697 infants. Multivariable logistic regression was used to investigate the effect of timing of resumption of maternal employment and maternal employment status on breastfeeding at 6 months postpartum after adjustment for maternal education, maternal age, maternal smoking during pregnancy and socioeconomic status of the child's area of residence. RESULTS: Fewer women employed full-time were breastfeeding their infants at 6 months (39%) than nonemployed women (56%). Participation in full-time employment before 6 months had a strong, negative effect on the likelihood of continuing breastfeeding for 6 months, adjusted OR = 0.35 (95%CI: 0.22-0.55). Compared to nonemployed women, fewer women in part-time employment were breastfeeding at 6 months (44%), adjusted OR = 0.49 (95% CI: 0.37 0.64). CONCLUSIONS: Results from this large representative cohort of Australian infants confirm that maternal employment in the first 6 months of life contributes to premature cessation of breastfeeding even when known risk factors for breastfeeding cessation are controlled for. PMID- 18394108 TI - The effect of alcohol intake on breastfeeding duration in Australian women. AB - AIM: This study investigated the relationships between alcohol consumption and breastfeeding initiation and duration. METHODS: DESIGN AND SETTING: A 12-month longitudinal study was conducted in two public hospitals in Perth, Australia between September 2002 and July 2003. INTERVENTION: Participating mothers completed a self-administered baseline questionnaire. Follow-up telephone interviews were conducted at 4, 10, 16, 22, 32, 40 and 52 weeks. MAIN OUTCOME MEASURES: Association of the relationships between alcohol consumption and breastfeeding initiation and duration. RESULTS: After 6 months of follow-up, women who consumed alcohol at levels of more than two standard drinks per day were almost twice as likely to discontinue breastfeeding than women who drank below these levels (HR 1.9, 95% CI 1.1, 3.0). CONCLUSION: Consuming alcohol in excess of two standard drinks per day during lactation was found to be independently associated with shorter breastfeeding duration, even after consideration of previously identified predictors of breastfeeding duration. Guidelines that provide direction on safe alcohol consumption for lactating mothers may help support extended breastfeeding duration. PMID- 18394109 TI - Self-reported violence amongst adolescents in Denmark: is alcohol a serious risk factor? AB - AIM: To describe the prevalence of adolescents' exposure to different types of violence (at home and outside the home) and associations between severe violence and alcohol consumption, taking account of gender and the influence of other factors. METHODS: A multimedia computer-based survey amongst a nationally representative sample of 9th grade pupils included comprehensive data on exposure to physical violence, use of alcohol and additional socio-demographic and behavioural factors. Analyses included gender disaggregated cross tabulations and logistic regressions. RESULTS: Around 9.3% of girls and 12.3% of boys reported being victims of physical violence within the last 12 months. Also, 2.7% of girls and 3.5% of boys reported severe violence. Gender differences existed in place of occurrence; girls were more frequently assaulted at home, boys outside the home. Alcohol was only associated with violence exposure outside the home for boys, not girls. CONCLUSIONS: About one-tenth of adolescents in Denmark reported exposure to any physical violence and 3% to severe violence during the last year. Alcohol consumption was not a risk factor for girls and only a risk factor for boys regarding violence occurring outside the home. PMID- 18394110 TI - On intimacy, sexual activities and exposure to sexual abuse among children and adolescents with mobility impairment. AB - AIM: The aim was to describe experiences of intimacy and sexual activity and exposure to sexual abuse among children and adolescents with mobility impairment, and to relate these experiences to socio-demographic data, disability characteristics and well-being. METHODS: This study included semi-structured interviews with 141 children and adolescents aged 7-18 years with mobility impairment. Interpersonal experiences of intimacy and sexuality, socio demographic data, disability characteristics and well-being were registered. RESULTS: About half of the children and adolescents in the study had been in a boy- or a girlfriend relationship, and about a fifth had an ongoing relationship. Of the adolescents, 15% had at least one experience of a sexual relationship. Whereas no particular sexual dysfunction was reported, 15% had concerns about their future sexual activities, presumably related to mobility impairment. A history of sexual abuse was reported by 7% in the age cohort of 13-18 years. The socio-demographic and disability-related features had a marginal influence on the experiences of intimacy and sexual activities. CONCLUSION: Several aspects of sexual health are not fully realized for children and adolescents with impaired mobility, and there is a need for specialized sexual health care services to protect the sexual rights of this group. PMID- 18394111 TI - Changes in physical activity in adolescent girls: a latent growth modelling approach. AB - AIM: To examine: (1) the developmental trajectory of physical activity as well the influence of body mass index (BMI) and friend support on initial physical activity and change of physical activity; and (2) the stability of physical activity in adolescent girls. METHODS: Participants were 193 urban adolescent girls and their best friends. Physical activity was measured using the 3-day physical activity recall. Best friend social support was assessed using a questionnaire. BMI was calculated based on height and body mass of the participants. Data were collected on four occasions over a 1.75-year period. RESULTS: The stability coefficients for moderate, vigorous and total physical activity were statistically significant and ranged from 0.25 to 0.62. There was a curvilinear decrease in physical activity and a curvilinear increase in BMI and friend social support across four measurement occasions. Latent growth modelling (LGM) revealed that initially higher level of friend social support was associated with higher physical activity, and higher level of BMI was associated with lower levels of physical activity. With the physical activity change factor, there were statistically significant and positive direct effect from friend social support change factor, and statistically significant and negative direct effect from BMI change factor. CONCLUSION: Change in friend social support was positively and change in BMI was inversely associated with a change in adolescent girls' physical activity. PMID- 18394112 TI - Cognitive and psychosocial development of children with Pierre Robin sequence. AB - AIM: It is unclear whether cognitive impairment in Pierre Robin sequence (PRS) results from a primary disturbance affecting both the brain and the mandible or from recurrent upper airway obstruction (UAO). If the latter were true, cognitive impairment should be preventable by early treatment of UAO. We wanted to determine the cognitive and psychosocial outcome of children with PRS treated with a new device aimed at relieving UAO in infancy (pre-epiglottic baton plate). METHODS: Thirty-four children with PRS and 34 healthy controls aged 4-11 years completed the Kaufman-Assessment Battery for Children (K-ABC) and a self-concept inventory. Parents rated their children's emotional and behavioural problems. Multi- and univariate analyses of covariance were performed, controlling for gender, age, parental education, family income and parental depression. RESULTS: The cognitive development of the PR children was within the reference range. Compared to healthy children, however, the children with PRS performed significantly poorer. There were no significant differences concerning self concept, emotional or behavioural problems. CONCLUSION: These children with non syndromic PRS who had received treatment of UAO in infancy performed worse in the K-ABC. However, this did not reflect a major cognitive impairment. PMID- 18394113 TI - Baby swimming and respiratory health. AB - AIM: To estimate the effect of baby swimming in the first 6 months of life on respiratory diseases from 6 to 18 months. METHODS: We used data from The Norwegian Mother and Child Cohort Study (MoBa) conducted by the Norwegian Institute of Public Health in children born between 1999 and 2005 followed from birth to the age of 18 months (n = 30,870). Health outcomes: lower respiratory tract infections (LRTI), wheeze and otitis media between 6 and 18 months of age. EXPOSURE: baby swimming at the age of 6 months. The effect of baby swimming was estimated by logistic regression analysis adjusting for potential confounders. RESULTS: About 25% of the children participated in baby swimming. The prevalence of LRTI was 13.3%, wheeze 40.0% and otitis media 30.4%. Children who were baby swimming were not more likely to have LRTI, to wheeze or to have otitis media. However, children with atopic mothers who attended baby swimming had an increased risk of wheeze, adjusted odds ratios (aOR) 1.24 (95% CI 1.11, 1.39), but not LRTI or otitis media. This was also the case for children without respiratory diseases before 6 months aOR 1.08 (95%CI 1.02-1.15). CONCLUSION: Baby swimming may be related to later wheeze. However, these findings warrant further investigation. PMID- 18394114 TI - Neonatal infective endarteritis complicating patent ductus arteriosus. AB - Infective endarteritis complicating patent ductus arteriosus (PDA) is a rare occurrence. Most cases are reported in older children and adults, and there are little published data describing this condition in the preterm neonate. We outline the presentation, clinical course, management and outcome of two affected infants born at less than 27 weeks of gestation. CONCLUSION: This report highlights the importance of a detailed cardiac evaluation in a neonate with persisting bacteraemia. An increasing awareness of neonatal ductal endarteritis combined with the wider availability of more sensitive echocardiography has implications for local neonatal service delivery. PMID- 18394115 TI - Co-occurrence of vitamin D-dependent rickets type 1 and phenylketonuria. AB - Vitamin D-dependent rickets type 1 (VDDR1) was diagnosed in a 15-month-old girl with well-controlled phenylketonuria (PKU). The patient was homozygous for the PAH mutation L249F. The PAH and CYP27B1 genes are both located on the long arm of chromosome 12 and could possibly have been inherited from a common ancestor. The parents were not aware of any ancestral relationship and the patient was compound heterozygous for two different CYP27B1 mutations (R389H and S416X). Her mutations were shown to originate from each of her four grandparents. In Norway, the co occurrence of PKU and VDDR1 is expected to occur by chance one to two times per billion births. CONCLUSION: The extremely rare co-occurrence of VDDR1 and PKU requires careful genetic work-up and close attention to family information, but the combined treatment of the two metabolic disorders may not create special problems. PMID- 18394116 TI - Bladder wall telangiectasis causing life-threatening haematuria in ataxia telangiectasia: a new observation. AB - We report two cases of life-threatening haemorrhage from bladder telangiectasia in children with ataxia-telangiectasia (A-T) who had been treated for lymphoma earlier in life. Whilst oculocutaneous telangiectasiae are an almost universal finding in this syndrome, bladder wall telangiectasis has not been reported previously. Both teenagers presented with recurrent severe haematuria due to extensive bladder telangiectasis. Recurrent haemorrhage was controlled with cystoscopic diathermy treatment. As A-T is a DNA repair disorder, it is possible that chemotherapy-mediated damage to the bladder mucosa prompted the development of clinically significant telangiectasis in these patients. CONCLUSION: We advocate early cystoscopy for A-T patients who develop haematuria to investigate the cause, and cystodiathermy to pre-emptively treat developing lesions prior to haemodynamically significant haemorrhage. PMID- 18394117 TI - Respiratory exacerbations in childhood associated with compound heterozygosity Phe508del/Arg117His-7T of the cystic fibrosis transmembrane regulator gene. AB - Debate continues regarding the clinical implications for compound heterozygotes identified with Phe508del and Arg117His-7T mutations of the cystic fibrosis transmembrane regulator (CFTR) gene. We report respiratory exacerbations and airway culture of Staphylococcus aureus and Pseudomonas aeruginosa in a child with this genotype. CONCLUSION: The compound heterozygote cystic fibrosis (CF) mutation Phe508del with Arg117His-7T should not necessarily be considered benign in childhood. PMID- 18394118 TI - The discovery of the structure and function of chromosomes: the basis of cytogenetics. PMID- 18394119 TI - The evolution of paediatrics from archaeological times to the mid-nineteenth century and the historical influence on present day practice. AB - The history of childcare dates back to the beginning of time. This article critically analyses the history of paediatrics from its roots to mid-nineteenth century with the view to examine its evolution and influence on today's practice. Paediatrics as a sub-speciality of medicine only began in the fifteenth century when the Four Incunabula were published in the West. This was the first attempt at producing a comprehensive and accessible reference paediatric text. However, long before the Incunabula, early traces of childcare are found in different cultures like Egyptian, Indian and Chinese. Modern paediatrics is a highly advanced field of medicine that relies on many recent technological innovations. In spite of these, this paper concludes that paediatrics today is very much based on concepts, such as observation and clinical skills, introduced many centuries ago. The basic approach used in everyday clinical practice owes more to century old ideas of scientists. PMID- 18394122 TI - The many faces of severe asthma. PMID- 18394124 TI - Investigation of the hygiene hypothesis: current issues and future directions. PMID- 18394123 TI - Treatment of exercise-induced asthma, respiratory and allergic disorders in sports and the relationship to doping: Part II of the report from the Joint Task Force of European Respiratory Society (ERS) and European Academy of Allergy and Clinical Immunology (EAACI) in cooperation with GA(2)LEN. AB - AIM: The aims of part II is to review the current recommended treatment of exercise-induced asthma (EIA), respiratory and allergic disorders in sports, to review the evidence on possible improvement of performance in sports by asthma drugs and to make recommendations for their treatment. METHODS: The literature cited with respect to the treatment of exercise induced asthma in athletes (and in asthma patients) is mainly based upon the systematic review given by Larsson et al. (Larsson K, Carlsen KH, Bonini S. Anti-asthmatic drugs: treatment of athletes and exercise-induced bronchoconstriction. In: Carlsen KH, Delgado L, Del Giacco S, editors. Diagnosis, prevention and treatment of exercise-related asthma, respiratory and allergic disorders in sports. Sheffield, UK: European Respiratory Journals Ltd, 2005:73-88) during the work of the Task Force. To assess the evidence of the literature regarding use of beta(2)-agonists related to athletic performance, the Task Force searched Medline for relevant papers up to November 2006 using the present search words: asthma, bronchial responsiveness, exercise-induced bronchoconstriction, athletes, sports, performance and beta(2)-agonists. Evidence level and grades of recommendation were assessed according to Sign criteria. RESULTS: Treatment recommendations for EIA and bronchial hyper-responsiveness in athletes are set forth with special reference to controller and reliever medications. Evidence for lack of improvement of exercise performance by inhaled beta(2)-agonists in healthy athletes serves as a basis for permitting their use. There is a lack of evidence of treatment effects of asthma drugs on EIA and bronchial hyper-responsiveness in athletes whereas extensive documentation exists in treatment of EIA in patients with asthma. The documentation on lack of improvement on performance by common asthma drugs as inhaled beta(2)-agonists with relationship to sports in healthy individuals is of high evidence, level (1+). CONCLUSIONS: Exercise induced asthma should be treated in athletes along same principles as in ordinary asthma patients with relevance to controller and reliever treatment after careful diagnosis. There is very high level of evidence for the lack of improvement in athletic performance by inhaled beta2-agonists. PMID- 18394125 TI - Carbohydrate-based particles reduce allergic inflammation in a mouse model for cat allergy. AB - BACKGROUND: Allergen-specific immunotherapy (ASIT) is the only treatment of allergic disease that gives long-lasting relief of symptoms. However, concerns for safety and efficiency have highlighted the need for improvement of the therapy. We have previously suggested carbohydrate-based particles (CBPs) as a novel adjuvant and allergen carrier for ASIT. Our aim of this study was to evaluate the therapeutic potential of CBPs in ASIT, employing a mouse model for cat allergy. METHODS: BALB/c mice were subcutaneously immunized with the recombinant (r) cat allergen Fel d 1 followed by intranasal challenge with cat dander extract (CDE). The sensitized mice were therapeutically treated with rFel d 1 covalently coupled to CBPs (CBP-rFel d 1). Airway hyper-reactivity (AHR), infiltration of leucocytes in bronchoalveolar lavage (BAL) fluid, allergen specific serum immunoglobulin levels and in vitro splenocyte responses were evaluated. RESULTS: Mice treated with CBP-rFel d 1 showed reduced features of allergic inflammation. They responded with (i) significantly decreased AHR and infiltration of eosinophils in BAL fluid after CDE challenge, (ii) the serum level of rFel d 1-specific IgE was reduced and the level of IgG(2)a was more pronounced after CBP-rFel d 1 treatment, and (iii) there was also a tendency of decreased allergen-specific cellular response. CONCLUSIONS: Carbohydrate-based particles are effective tools as adjuvant and allergen carriers for use in ASIT and constitutes a promising strategy to improve allergy treatment. PMID- 18394126 TI - Methacholine vs adenosine on intra and extrathoracic airway hyperresponsiveness in patients with cough variant asthma. AB - BACKGROUND: Airway hyperresponsiveness (AHR) can be studied by bronchoprovocation test (BPT) using direct (methacholine - MCh) or indirect (adenosine 5' monophosphate - AMP) stimuli. These two substances have not been compared in cough variant asthma (CVA). OBJECTIVE: We designed a randomized, single-blind, cross-over study to compare AMP and MCh in the detection of CVA. Additionally, we examined whether assessment of extrathoracic airway hyperresponsiveness (EAHR) during MCh and AMP helped in the evaluation of CVA. METHODS: Patients with CVA with previous positive MCh BPT performed challenges with AMP and MCh. The variables were: (i) a provocative dose producing a 20% fall in forced expiratory volume in 1 s (FEV(1)) value (PD(20)MCh); (ii) a provocative dose producing a 25% fall in the maximal mid-inspiratory flow (FIF(50)) from baseline (PD(25)MCh) for MCh; (iii) a provocative concentration producing a 20% fall in FEV(1) value (PC(20)AMP) and (iv) a provocative concentration producing a 25% fall in the FIF(50) from baseline (PC(25)AMP) for AMP. RESULTS: All 113 patients with CVA responded to PD(20)MCh and 96% and 69% responded to PC(20)AMP, if we used PC(20) /= 30) and factors related with asthma severity in patients with difficult-to-treat asthma. METHODS: One hundred and thirty-six nonsmoking asthmatic adults with persistent symptoms despite high doses of inhaled or oral corticosteroids and long-acting bronchodilators were studied [70% female, median (range) age 44.6 (18-75) years, 32% on daily oral corticosteroids]. The association between obesity, lung function parameters [forced expiratory volume in 1 s (FEV(1)), functional residual capacity/total lung capacity (FRC/TLC)], inflammatory markers [blood eosinophils, sputum eosinophils and neutrophils, exhaled nitric oxide (FE(NO)), airway hyperresponsiveness, C-reactive protein (CRP)] and aggravating co-morbid factors (severe chronic sinus disease, gastro-esophageal reflux, recurrent respiratory infections, psychopathology and obstructive sleep apnea) was investigated. RESULTS: Obese patients (n = 29) had a higher FEV(1)%pred (P = 0.05) and a lower FRC/TLC%pred (P < 0.01) compared with nonobese patients (n = 107). Body mass index was inversely related with sputum eosinophils (r = -0.36, P < 0.01) and FE(NO) (r = -0.30, P < 0.01). Obese patients had an increased risk for gastro esophageal reflux (OR = 2.3) and sleep apnea (OR = 3.1). CONCLUSION: Obesity in patients with difficult-to-treat asthma is inversely related with sputum eosinophils and FE(NO), and positively associated with the presence of co-morbid factors and reduced lung volumes. This suggests that other factors than airway inflammation alone explain the relationship between obesity and asthma severity. PMID- 18394132 TI - Association of obesity and insulin resistance with asthma and aeroallergen sensitization. AB - BACKGROUND: It has been hypothesized that obesity and insulin resistance may play a role in the development of asthma and allergy. The aim of the study was to examine the association of obesity and insulin resistance with asthma and aeroallergen sensitization. METHODS: Cross-sectional population-based study of 3609 Danish men and women aged 30-60 years. Aeroallergen sensitization was defined as positive levels of specific IgE against a panel of inhalant allergens. Asthma was defined as self-reported physician diagnosed asthma. Allergic asthma was defined as the presence of both asthma and aeroallergen sensitization. The homeostasis model assessment of insulin resistance was used to estimate the degree of insulin resistance. Body mass index, waist-to-hip ratio, and waist circumference were used as measures of obesity. Data were analyzed by multiple logistic regression analyses. RESULTS: Obesity was associated with increased risk of aeroallergen sensitization as well as allergic and nonallergic asthma. Insulin resistance was asssociated with aeroallergen sensitization and allergic asthma, but not nonallergic asthma. The associations of obesity with aeroallegen sensitization and allergic asthma became nonsignificant after adjustment for insulin resistance, whereas the association of obesity with nonallergic asthma was unaffected. No sex-differences were observed. CONCLUSION: Obesity may be related to an increased risk of aeroallergen sensitization and allergic asthma through mechanisms also involved in the development of insulin resistance. PMID- 18394133 TI - Inflammation and functional outcome in diisocyanate-induced asthma after cessation of exposure. AB - BACKGROUND: The clinical outcome of diisocyanate-induced asthma has been found to be poor despite cessation of exposure. Our aim was to study the outcome of diisocyanate-induced asthma after initiation of inhaled steroid treatment at a mean period of 7 months (range 2-60 months) after cessation of exposure by following up lung function and bronchial inflammation. METHODS: Bronchoscopy was performed on 17 patients 2 days after a positive inhalation challenge test, after which budesonide 1600 mug a day was started. Bronchoscopy, spirometry, and histamine challenge tests were repeated at 6 months and on average 3 years. The results were also compared with those obtained from 15 healthy control subjects. RESULTS: Nonspecific bronchial hyperreactivity diminished significantly (P = 0.006); however, forced expiratory volume in 1 s (FEV1) and forced vital capacity (FVC) values decreased, with a median yearly reduction of FEV1 of 79 ml. The count of mast cells in bronchial mucosa decreased (P = 0.012) and that of macrophages increased (P = 0.001). Interleukin-4 level in mucosa was during the first year significantly higher than in controls but its level decreased in the follow-up. Interleukin-6, interleukin-15, and tumour necrosis factor alpha messenger-RNA levels were significantly higher in hyperreactive patients than in nonhyperreactive patients at the end of the follow-up. CONCLUSION: Our results indicate that inflammation may persist in diisocyanate-induced asthma despite inhaled steroid medication. However, TH2-type inflammation diminished. Persistent nonspecific bronchial hyperreactivity was associated with proinflammatory acting cytokines produced mainly by macrophages. Considering the poor prognosis of the disease the findings could be utilized to develop the follow-up and treatment of diisocyanate-induced asthma. PMID- 18394134 TI - Thresholds for food allergens and their value to different stakeholders. AB - Thresholds constitute a critical piece of information in assessing the risk from allergenic foods at both the individual and population levels. Knowledge of the minimum dose that can elicit a reaction is of great interest to all food allergy stakeholders. For allergic individuals and health professionals, individual threshold data can inform allergy management. Population thresholds can help both the food industry and regulatory authorities assess the public health risk and design appropriate food safety objectives to guide risk management. Considerable experience has been gained with the double-blind placebo-controlled food challenge (DBPCFC), but only recently has the technique been adapted to provide data on thresholds. Available data thus vary greatly in quality, with relatively few studies providing the best quality individual data, using the low-dose DBPCFC. Such high quality individual data also form the foundation for population thresholds, but these also require, in addition to an adequate sample size, a good characterization of the tested population in relation to the whole allergic population. Determination of thresholds at both an individual level and at a population level is influenced by many factors. This review describes a low-dose challenge protocol developed as part of the European Community-funded Integrated Project Europrevall, and strongly recommends its wider use so that data are generated that can readily increase the power of existing studies. PMID- 18394136 TI - Important questions in Allergy: 1--drug allergy/hypersensitivity. PMID- 18394137 TI - Improving latex-allergy diagnosis: the clinical role of Hev b8-specific IgE. PMID- 18394138 TI - Asthma-related mortality in France, 1980-2005: decline since the last decade. PMID- 18394135 TI - Gender-medicine aspects in allergology. AB - Despite the identical immunological mechanisms activating the release of mediators and consecutive symptoms in immediate-type allergy, there is still a clear clinical difference between female and male allergic patients. Even though the risk of being allergic is greater for boys in childhood, almost from adolescence onwards it seems to be a clear disadvantage to be a woman as far as atopic disorders are concerned. Asthma, food allergies and anaphylaxis are more frequently diagnosed in females. In turn, asthma and hay fever are associated with irregular menstruation. Pointing towards a role of sex hormones, an association of asthma and intake of contraceptives, and a risk for asthma exacerbations during pregnancy have been observed. Moreover, peri- and postmenopausal women were reported to increasingly suffer from asthma, wheeze and hay fever, being even enhanced by hormone replacement therapy. This may be on account of the recently identified oestradiol-receptor-dependent mast-cell activation. As a paradox of nature, women may even become hypersensitive against their own sex hormones, resulting in positive reactivity upon intradermal injection of oestrogen or progesterone. More importantly, this specific hypersensitivity is associated with recurrent miscarriages. Even though there is a striking gender-specific bias in IgE-mediated allergic diseases, public awareness of this fact still remains minimal today. PMID- 18394139 TI - Training does not affect exhaled nitric oxide in competitive swimmers. PMID- 18394140 TI - Selective hypersensitivity to nimesulide. PMID- 18394141 TI - FCER1A gene exon 1A polymorphisms in Japanese and Polish subjects--a comparative analysis of haplotypes. PMID- 18394142 TI - Development of allergic asthma in an asthmatic recipient of a nonatopic lung under systemic immunosuppression. PMID- 18394144 TI - Glycan microarray analysis of Candida glabrata adhesin ligand specificity. AB - The Candida glabrata genome encodes at least 23 members of the EPA (epithelial adhesin) family responsible for mediating adherence to host cells. To better understand the mechanism by which the Epa proteins contribute to pathogenesis, we have used glycan microarray analysis to characterize their carbohydrate-binding specificities. Using Saccharomyces cerevisiae strains surface-expressing the N terminal ligand-binding domain of the Epa proteins, we found that the three Epa family members functionally identified as adhesins in Candida glabrata (Epa1, Epa6 and Epa7) bind to ligands containing a terminal galactose residue. However, the specificity of the three proteins for glycans within this class varies, with Epa6 having a broader specificity range than Epa1 or Epa7. This result is intriguing given the close homology between Epa6 and Epa7, which are 92% identical at the amino acid level. We have mapped a five-amino-acid region within the N-terminal ligand-binding domain that accounts for the difference in specificity of Epa6 and Epa7 and show that these residues contribute to adherence to both epithelial and endothelial cell lines in vitro. PMID- 18394145 TI - Selective activation by the guanine nucleotide exchange factor Don1 is a main determinant of Cdc42 signalling specificity in Ustilago maydis. AB - The highly conserved GTP-binding proteins Cdc42 and Rac1 regulate cytokinesis, establishment of cell polarity and vesicular trafficking. In the dimorphic fungus Ustilago maydis, Rac1 is required for cell polarity and budding, while Cdc42 is essential for cell separation during cytokinesis. The same cell separation defect is also observed in mutants that lack Don1, a guanine nucleotide exchange factor (GEF) of the Dbl family. We have generated a series of chimeric GTP-binding proteins consisting of different portions of Cdc42 and Rac1. In vivo complementation analysis revealed that a short region encompassing amino acids 41 56 determines signalling specificity. Remarkably, substitution of a single amino acid at position 56 within this specificity domain is sufficient to confer Cdc42 function to Rac1 in vivo. Expression of Rac1(W56F) in Delta cdc42 mutant cells resulted in complementation of the cell separation defect. In vitro GDP/GTP exchange assays demonstrated that the Dbl family GEF Don1 is highly specific for Cdc42 and cannot activate Rac1. However, if Rac1(W56F) is used as a substrate, Don1 is able to stimulate GDP/GTP exchange. Together these data indicate that activation by the GEF Don1 is an important determinant of Cdc42-specific signalling in vivo. PMID- 18394146 TI - Dissimilatory iron reduction in Escherichia coli: identification of CymA of Shewanella oneidensis and NapC of E. coli as ferric reductases. AB - Over geological time scales, microbial reduction of chelated Fe(III) or Fe(III) minerals has profoundly affected today's composition of our bio- and geosphere. However, the electron transfer reactions that are specific and defining for dissimilatory iron(III)-reducing (DIR) bacteria are not well understood. Using a synthetic biology approach involving the reconstruction of the putative electron transport chain of the DIR bacterium Shewanella oneidensis MR-1 in Escherichia coli, we showed that expression of cymA was necessary and sufficient to convert E. coli into a DIR bacterium. In intact cells, the Fe(III)-reducing activity was limited to Fe(III) NTA as electron acceptor. In vitro biochemical analysis indicated that CymA, which is a cytoplasmic membrane-associated tetrahaem c-type cytochrome, carries reductase activity towards Fe(III) NTA, Fe(III) citrate, as well as to AQDS, a humic acid analogue. The in vitro specific activities of Fe(III) citrate reductase and AQDS reductase of E. coli spheroplasts were 10x and 30x higher, respectively, relative to the specific rates observed in intact cells, suggesting that access of chelated and insoluble forms of Fe(III) and AQDS is restricted in whole cells. Interestingly, the E. coli CymA orthologue NapC also carried ferric reductase activity. Our data support the argument that the biochemical mechanism of Fe(III) reduction per se was not the key innovation leading to environmental relevant DIR bacteria. Rather, the evolution of an extension of the electron transfer pathway from the Fe(III) reductase CymA to the cell surface via a system of periplasmic and outer membrane cytochrome proteins enabled access to diffusion-impaired electron acceptors. PMID- 18394147 TI - Novel coiled-coil cell division factor ZapB stimulates Z ring assembly and cell division. AB - Formation of the Z ring is the first known event in bacterial cell division. However, it is not yet known how the assembly and contraction of the Z ring are regulated. Here, we identify a novel cell division factor ZapB in Escherichia coli that simultaneously stimulates Z ring assembly and cell division. Deletion of zapB resulted in delayed cell division and the formation of ectopic Z rings and spirals, whereas overexpression of ZapB resulted in nucleoid condensation and aberrant cell divisions. Localization of ZapB to the divisome depended on FtsZ but not FtsA, ZipA or FtsI, and ZapB interacted with FtsZ in a bacterial two hybrid analysis. The simultaneous inactivation of FtsA and ZipA prevented Z ring assembly and ZapB localization. Time lapse microscopy showed that ZapB-GFP is present at mid-cell in a pattern very similar to that of FtsZ. Cells carrying a zapB deletion and the ftsZ84(ts) allele exhibited a synthetic sick phenotype and aberrant cell divisions. The crystal structure showed that ZapB exists as a dimer that is 100% coiled-coil. In vitro, ZapB self-assembled into long filaments and bundles. These results raise the possibility that ZapB stimulates Z ring formation directly via its capacity to self-assemble into larger structures. PMID- 18394148 TI - Bacitracin sensing in Bacillus subtilis. AB - The extracellular presence of antibiotics is a common threat in microbial life. Their sensitive detection and subsequent induction of appropriate resistance mechanisms is therefore a prerequisite for survival. The bacitracin stress response network of Bacillus subtilis consists of four signal-transducing systems, the two-component systems (TCS) BceRS, YvcPQ and LiaRS, and the extracytoplasmic function (ECF) sigma factor sigma(M). Here, we investigated the mechanism of bacitracin perception and the response hierarchy within this network. The BceRS-BceAB TCS/ABC transporter module is the most sensitive and efficient bacitracin resistance determinant. The ABC transporter BceAB not only acts as a bacitracin detoxification pump, but is also crucial for bacitracin sensing, indicative of a novel mechanism of stimulus perception, conserved in Firmicutes bacteria. The Bce system seems to respond to bacitracin directly (drug sensing), whereas the LiaRS TCS and sigma(M) respond only at higher concentrations and indirectly to bacitracin action (damage sensing). The YvcPQ YvcRS system is subject to cross-activation via the paralogous Bce system, and is therefore only indirectly induced by bacitracin. The bacitracin stress response network is optimized to respond to antibiotic gradients in a way that maximizes the gain and minimizes the costs of this stress response. PMID- 18394149 TI - Pneumopericardium should be considered with electrocardiogram changes after blunt chest trauma: a case report. AB - INTRODUCTION: Electrocardiogram (ECG) abnormalities in patients with blunt chest trauma are diverse and non-specific, but may be indicative of potentially life threatening conditions. CASE PRESENTATION: We report a rare case of pneumopericardium with extreme ECG abnormalities after blunt chest trauma in a 22 year-old male. The diagnosis was confirmed using computed tomography (CT) scanning. The case is discussed, together with its differential diagnosis and the aetiology of pneumopericardium and tension pneumopericardium. CONCLUSION: Pneumopericardium should be distinguished from other pathologies such as myocardial contusion and myocardial infarction because of the possible development of tension pneumopericardium. Early CT scanning is important in the evaluation of blunt chest trauma. PMID- 18394150 TI - Correcting biases in psychiatric diagnostic practice in Northwest Russia: comparing the impact of a general educational program and a specific diagnostic training program. AB - BACKGROUND: A general education in psychiatry does not necessary lead to good diagnostic skills. Specific training programs in diagnostic coding are established to facilitate implementation of ICD-10 coding practices. However, studies comparing the impact of these two different educational approaches on diagnostic skills are lacking. The aim of the current study was to find out if a specific training program in diagnostic coding improves the diagnostic skills better than a general education program, and if a national bias in diagnostic patterns can be minimised by a specific training in diagnostic coding. METHODS: A pre post design study with two groups was carried in the county of Archangelsk, Russia. The control group (39 psychiatrists) took the required course (general educational program), while the intervention group (45 psychiatrists) were given a specific training in diagnostic coding. Their diagnostic skills before and after education were assessed using 12 written case-vignettes selected from the entire spectrum of psychiatric disorders. RESULTS: There was a significant improvement in diagnostic skills in both the intervention group and the control group. However, the intervention group improved significantly more than did the control group. The national bias was partly corrected in the intervention group but not to the same degree in the control group. When analyzing both groups together, among the background factors only the current working place impacted the outcome of the intervention. CONCLUSION: Establishing an internationally accepted diagnosis seems to be a special skill that requires specific training and needs to be an explicit part of the professional educational activities of psychiatrists. It does not appear that that skill is honed without specific training. The issue of national diagnostic biases should be taken into account in comparative cross-cultural studies of almost any character. The mechanisms of such biases are complex and need further consideration in future research. Future research should also address the question as to whether the observed improvement in diagnostic skills after specific training actually leads to changes in routine diagnostic practice. PMID- 18394151 TI - OppA, the ecto-ATPase of Mycoplasma hominis induces ATP release and cell death in HeLa cells. AB - BACKGROUND: In the facultative human pathogen Mycoplasma hominis, which belongs to the cell wall-less Mollicutes, the surface-localised substrate-binding domain OppA of the oligopeptide permease was characterised as the main ecto-ATPase. RESULTS: With the idea that extra-cellular ATP could only be provided by the infected host cells we analysed the ATP release of HeLa cells after incubation with different preparations of Mycoplasma hominis: intact bacterial cells, the membrane fraction with or without OppA, recombinant OppA as well as an ATPase deficient OppA mutant. Release of ATP into the supernatant of the HeLa cells was primarily determined in all samples lacking ecto-ATPase activity of OppA. In the presence of the ATPase inhibitor DIDS the amount of ATP in the OppA-containing samples increased. This increase was maximal after incubation with fractions containing OppA protein indicating that OppA is involved in ATP release and subsequent hydrolysis. Real-time PCR analyses revealed that the proliferation of HeLa cells is reduced after infection with M. hominis and flow cytometry experiments established that OppA induces greater apoptosis than necrosis of HeLa cells whereas the preservation of ecto-ATPase activity of OppA induces apoptosis. CONCLUSION: The OppA induced ATP-release and -hydrolysis induced cell death of M. hominis infected HeLa cells was predominantly due to apoptosis rather than necrosis. Future work will elucidate whether the induction of apoptosis is indispensable for survival of these non-invasive pathogen. PMID- 18394152 TI - A comparison of four scoring methods based on the parent-rated Strengths and Difficulties Questionnaire as used in the Dutch preventive child health care system. AB - BACKGROUND: Validated questionnaires can support the identification of psychosocial problems by the Preventive Child Health Care (PCH) system. This study assesses the validity and added value of four scoring methods used with the Strengths and Difficulties Questionnaire (SDQ) for the identification of psychosocial problems among children aged 7-12 by the PCH. METHODS: We included 711 (of 814) children (response: 87%) aged 7-12 undergoing routine health assessments in nine PCH services across the Netherlands. Child health professionals interviewed and examined children and parents. Prior to the interview, parents completed the SDQ and the Child Behaviour Checklist (CBCL), which were not shown to the professionals. The CBCL and data about the child's current treatment status were used as criteria for the validity of the SDQ. We used four SDQ scoring approaches: an elevated SDQ Total Difficulties Score (TDS), parent-defined difficulties, an elevated score for emotional symptoms, conduct problems or hyperactivity in combination with a high impairment score, and a combined score: an elevated score for any of these three methods. RESULTS: The Cohen's Kappa ranged from 0.33 to 0.64 for the four scoring methods with the CBCL scores and treatment status, generally indicating a moderate to good agreement. All four methods added significantly to the identification of problems by the PCH. Classification based on the TDS yielded results similar to more complicated methods. CONCLUSION: The SDQ is a valid tool for the identification of psychosocial problems by PCH. As a first step, the use of a simple classification based on the SDQ TDS is recommended. PMID- 18394153 TI - Hydrogenated fat diet intake during pregnancy and lactation modifies the PAI-1 gene expression in white adipose tissue of offspring in adult life. AB - We examine whether feeding pregnant and lactating rats hydrogenated fats rich in trans fatty acids modifies the plasma lipid profiles and the expression of adipokines involved with insulin resistance and cardiovascular disease in their 90-day-old offspring. Pregnant and lactating Wistar rats were fed with either a control diet (C group) or one enriched with hydrogenated vegetable fat (T group). Upon weaning, the male pups were sorted into four groups: CC, mothers were receiving C and pups were kept on C; CT, mothers were receiving C and pups were fed with T; TT, mothers were receiving T and pups were kept on T; TC, mothers were receiving T and pups were fed with C. Pups' food intake and body weight were quantified weekly and the pups were killed at day 90 of life by decapitation. Blood and carcass as well as retroperitoneal, epididymal, and subcutaneous white adipose tissues were collected. Food intake and body weight were lower in TC and TT, and metabolic efficiency was reduced in TT. Offspring of TT and TC rats had increased white adipose tissue PAI-1 gene expression. Insulin receptor was higher in TT than other groups. Ingestion of hydrogenated vegetable fat by the mother during gestation and lactation could promote deleterious consequences, even after the withdrawal of the causal factor. PMID- 18394154 TI - Multidimensional scaling for large genomic data sets. AB - BACKGROUND: Multi-dimensional scaling (MDS) is aimed to represent high dimensional data in a low dimensional space with preservation of the similarities between data points. This reduction in dimensionality is crucial for analyzing and revealing the genuine structure hidden in the data. For noisy data, dimension reduction can effectively reduce the effect of noise on the embedded structure. For large data set, dimension reduction can effectively reduce information retrieval complexity. Thus, MDS techniques are used in many applications of data mining and gene network research. However, although there have been a number of studies that applied MDS techniques to genomics research, the number of analyzed data points was restricted by the high computational complexity of MDS. In general, a non-metric MDS method is faster than a metric MDS, but it does not preserve the true relationships. The computational complexity of most metric MDS methods is over O(N2), so that it is difficult to process a data set of a large number of genes N, such as in the case of whole genome microarray data. RESULTS: We developed a new rapid metric MDS method with a low computational complexity, making metric MDS applicable for large data sets. Computer simulation showed that the new method of split-and-combine MDS (SC-MDS) is fast, accurate and efficient. Our empirical studies using microarray data on the yeast cell cycle showed that the performance of K-means in the reduced dimensional space is similar to or slightly better than that of K-means in the original space, but about three times faster to obtain the clustering results. Our clustering results using SC-MDS are more stable than those in the original space. Hence, the proposed SC-MDS is useful for analyzing whole genome data. CONCLUSION: Our new method reduces the computational complexity from O(N3) to O(N) when the dimension of the feature space is far less than the number of genes N, and it successfully reconstructs the low dimensional representation as does the classical MDS. Its performance depends on the grouping method and the minimal number of the intersection points between groups. Feasible methods for grouping methods are suggested; each group must contain both neighboring and far apart data points. Our method can represent high dimensional large data set in a low dimensional space not only efficiently but also effectively. PMID- 18394155 TI - Adolescents with metabolic syndrome have a history of low aerobic fitness and physical activity levels. AB - PURPOSE: Metabolic syndrome (MS) is a clustering of cardiovascular disease risk factors that identifies individuals with the highest risk for heart disease. Two factors that may influence the MS are physical activity and aerobic fitness. This study determined if adolescent with the MS had low levels of aerobic fitness and physical activity as children. METHODS: This longitudinal, exploratory study had 389 participants: 51% girls, 84% Caucasian, 12% African American, 1% Hispanic, and 3% other races, from the State of North Carolina. Habitual physical activity (PA survey), aerobic fitness (VO2max), body mass index (BMI), blood pressure, and lipids obtained at 7-10 y of age were compared to their results obtained 7 y later at ages 14-17 y. RESULTS: Eighteen adolescents (4.6%) developed 3 or more characteristics of the MS. Logistic regression, adjusting for BMI percentile, blood pressure, and cholesterol levels, found that adolescents with the MS were 6.08 (95%CI = 1.18-60.08) times more likely to have low aerobic fitness as children and 5.16 (95%CI = 1.06-49.66) times more likely to have low PA levels. CONCLUSION: Low levels of childhood physical activity and aerobic fitness are associated with the presence of the metabolic syndrome in adolescents. Thus, efforts need to begin early in childhood to increase exercise. PMID- 18394156 TI - Comparison of the developmental potential of 2-week-old preantral follicles derived from vitrified ovarian tissue slices, vitrified whole ovaries and vitrified/transplanted newborn mouse ovaries using the metal surface method. AB - BACKGROUND: Cryopreservation of preantral follicles or ovarian tissues would enable the storage of large numbers of primordial follicles or preantral follicles and preserves the structural integrity of somatic and reproductive cells. In the present study, we compared the developmental potential of cryopreserved two-week-old mouse preantral follicles, ovarian tissue slices, two week-old mouse ovaries and newborn mouse ovaries using a metal plate with a high cooling rate for cooling the droplet of vitrification solution. METHODS: Groups of 2 to 4 samples (including of 14-day old preantral follicles, ovarian tissue slices, whole ovaries, and whole newborn ovaries) were exposed to 4% ethylene glycol (EG) in DPBS + 10% FBS for 15 min and then rinsed in a vitrification solution composed of 6 M ethylene glycol and 0.4 M trehalose in DPBS + 10% FBS. Equilibration in room temperature was performed for 20-30 seconds for preantral follicle and 5 min equilibration was performed in an ice bath for ovaries. The samples were dropped onto the surface of metal plate around -180 degrees C in the volume of 2 microl and 6 microl. After thawing, the ovarian tissue was mechanically isolated for collecting the preantral follicles. The thawed newborn ovaries were transplanted under the renal capsule of recipient male mice for 14 days. Preantral follicles collected from each groups were cultured individually in 20-microl droplets of alpha-MEM culture medium in culture dish for 12 days. On the day 12 of culture, the cumulus-oocyte complexes (COCs) were collected for IVM and IVF. Fertilization and embryo cleavage were scored. RESULTS: After the vitrification of 14-day-old preantral follicles using 2 microl or 6 microl droplet onto surface of metal plate, the results indicated that no significant difference in survival rate, antral-like cavity formation, COCs collected, 2 cell embryo cleavage and blastocyst development was found in vitrification of the 2 microl and 6 microl droplet groups. As comparing 14-day old ovarian tissue (ovarian tissue slices and whole ovaries) and whole newborn ovaries vitrified in 6 microl droplet, lower success rates of antral-like cavity formation and COCs collection were found in the whole ovaries group. CONCLUSION: Our results suggest that the metal plate surface vitrification method is an appropriate and convenient method for cryopreservation of mouse ovaries and preantral follicles. The droplet volume of vitrification solution in 2 microl and 6 microl can be an option. PMID- 18394157 TI - Flow-volume loops derived from three-dimensional echocardiography: a novel approach to the assessment of left ventricular hemodynamics. AB - BACKGROUND: This study explores the feasibility of non-invasive evaluation of left ventricular (LV) flow-volume dynamics using 3-dimensional (3D) echocardiography, and the capacity of such an approach to identify altered LV hemodynamic states caused by valvular abnormalities. METHODS: Thirty-one patients with moderate-severe aortic (AS) and mitral (MS) stenoses (21 and 10 patients, respectively) and 10 healthy volunteers underwent 3D echocardiography with full volume acquisition using Philips Sonos 7500 equipment. The digital 3D data were post- processed using TomTec software. LV flow-volume loops were subsequently constructed for each subject by plotting instantaneous LV volume data sampled throughout the cardiac cycle vs. their first derivative representing LV flow. After correction for body surface area, an average flow-volume loop was calculated for each subject group. RESULTS: Flow-volume loops were obtainable in all subjects, except 3 patients with AS. The flow-volume diagrams displayed clear differences in the form and position of the loops between normal individuals and the respective patient groups. In patients with AS, an "obstructive" pattern was observed, with lower flow values during early systole and larger end-systolic volume. On the other hand, patients with MS displayed a "restrictive" flow-volume pattern, with reduced diastolic filling and smaller end-diastolic volume. CONCLUSION: Non-invasive evaluation of LV flow-volume dynamics using 3D echocardiographic data is technically possible and the approach has a capacity to identify certain specific types of alteration of LV flow-volume pattern caused by valvular abnormalities, thus reflecting underlying hemodynamic states specific for these abnormalities. PMID- 18394158 TI - Histone deacetylase inhibition accelerates the early events of stem cell differentiation: transcriptomic and epigenetic analysis. AB - BACKGROUND: Epigenetic mechanisms regulate gene expression patterns affecting cell function and differentiation. In this report, we examine the role of histone acetylation in gene expression regulation in mouse embryonic stem cells employing transcriptomic and epigenetic analysis. RESULTS: Embryonic stem cells treated with the histone deacetylase inhibitor Trichostatin A (TSA), undergo morphological and gene expression changes indicative of differentiation. Gene profiling utilizing Affymetrix microarrays revealed the suppression of important pluripotency factors, including Nanog, a master regulator of stem cell identity, and the activation of differentiation-related genes. Transcriptional and epigenetic changes induced after 6-12 hours of TSA treatment mimic those that appear during embryoid body differentiation. We show here that the early steps of stem cell differentiation are marked by the enhancement of bulk activatory histone modifications. At the individual gene level, we found that transcriptional reprogramming triggered by histone deacetylase inhibition correlates with rapid changes in activating K4 trimethylation and repressive K27 trimethylation of histone H3. The establishment of H3K27 trimethylation is required for stable gene suppression whereas in its absence, genes can be reactivated upon TSA removal. CONCLUSION: Our data suggest that inhibition of histone deacetylases accelerates the early events of differentiation by regulating the expression of pluripotency- and differentiation-associated genes in an opposite manner. This analysis provides information about genes that are important for embryonic stem cell function and the epigenetic mechanisms that regulate their expression. PMID- 18394159 TI - Towards the development of Bacillus subtilis as a cell factory for membrane proteins and protein complexes. AB - BACKGROUND: The Gram-positive bacterium Bacillus subtilis is an important producer of high quality industrial enzymes and a few eukaryotic proteins. Most of these proteins are secreted into the growth medium, but successful examples of cytoplasmic protein production are also known. Therefore, one may anticipate that the high protein production potential of B. subtilis can be exploited for protein complexes and membrane proteins to facilitate their functional and structural analysis. The high quality of proteins produced with B. subtilis results from the action of cellular quality control systems that efficiently remove misfolded or incompletely synthesized proteins. Paradoxically, cellular quality control systems also represent bottlenecks for the production of various heterologous proteins at significant concentrations. CONCLUSION: While inactivation of quality control systems has the potential to improve protein production yields, this could be achieved at the expense of product quality. Mechanisms underlying degradation of secretory proteins are nowadays well understood and often controllable. It will therefore be a major challenge for future research to identify and modulate quality control systems of B. subtilis that limit the production of high quality protein complexes and membrane proteins, and to enhance those systems that facilitate assembly of these proteins. PMID- 18394161 TI - Folding and assembly of large macromolecular complexes monitored by hydrogen deuterium exchange and mass spectrometry. AB - Recent advances in protein mass spectrometry (MS) have enabled determinations of hydrogen deuterium exchange (HDX) in large macromolecular complexes. HDX-MS became a valuable tool to follow protein folding, assembly and aggregation. The methodology has a wide range of applications in biotechnology ranging from quality control for over-expressed proteins and their complexes to screening of potential ligands and inhibitors. This review provides an introduction to protein folding and assembly followed by the principles of HDX and MS detection, and concludes with selected examples of applications that might be of interest to the biotechnology community. PMID- 18394160 TI - Protein folding and conformational stress in microbial cells producing recombinant proteins: a host comparative overview. AB - Different species of microorganisms including yeasts, filamentous fungi and bacteria have been used in the past 25 years for the controlled production of foreign proteins of scientific, pharmacological or industrial interest. A major obstacle for protein production processes and a limit to overall success has been the abundance of misfolded polypeptides, which fail to reach their native conformation. The presence of misfolded or folding-reluctant protein species causes considerable stress in host cells. The characterization of such adverse conditions and the elicited cell responses have permitted to better understand the physiology and molecular biology of conformational stress. Therefore, microbial cell factories for recombinant protein production are depicted here as a source of knowledge that has considerably helped to picture the extremely rich landscape of in vivo protein folding, and the main cellular players of this complex process are described for the most important cell factories used for biotechnological purposes. PMID- 18394162 TI - Protein C: a potential biomarker in severe sepsis and a possible tool for monitoring treatment with drotrecogin alfa (activated). AB - INTRODUCTION: Drotrecogin alfa (activated; DrotAA) treatment, a 96-hour infusion, reduces 28-day mortality in severe sepsis to approximately 25%. The question remains whether a longer infusion or higher dose could increase rate of survival. The goal of this study was to identify a dependable, sensitive measure with which to monitor disease progression and response in patients during DrotAA treatment. METHODS: Data on severe sepsis patients included in PROWESS (placebo-controlled, double-blind, randomized study of 850 DrotAA and 840 placebo individuals) and ENHANCE (single-arm, open-label study of 2,375 DrotAA patients) studies were analyzed. In these studies, DrotAA (24 mug/kg per hour) or placebo was infused for 96 hours and patients were followed for 28 days. Data on six laboratory measures and five organ dysfunctions were systematically analyzed to identify a potential surrogate end-point for monitoring DrotAA therapy and predicting 28-day mortality at the end of therapy. To allow comparison across variables, sensitivity and specificity analyses identified cut-off values for preferred outcome, and relative risks for being above or below cut-offs were calculated, as was the 'proportion of treatment effect explained' (PTEE) to identify biomarkers that contribute to benefit from DrotAA. RESULTS: Protein C was the only variable that correlated with outcome across all analyses. Using placebo data, a baseline protein C under 40% was established as a useful predictor of outcome (odds ratio 2.12). Similar odds ratios were associated with cut-off values of other biomarkers, but the treatment benefit associated with DrotAA was significantly greater below the cut-off than above the cut-off only for protein C (relative risk for 28-day mortality 0.66 versus 0.88; P = 0.04). Protein C was the only end of-infusion biomarker that potentially explained at least 50% of the benefit from DrotAA (PTEE 57.2%). The PTEE was 41% for cardiovascular Sequential Organ Failure Assessment score and for d-dimer. At the end of infusion (day 4), protein C categories ( 80%) remained significantly related to mortality, regardless of treatment assignment. CONCLUSION: Based on systematic analyses of 11 variables measured in severe sepsis clinical trials, protein C was the only variable consistently correlated with both DrotAA treatment effect and survival. Further study is needed to determine whether longer infusions or higher doses of DrotAA would achieve the goal of normalizing protein C in more patients with severe sepsis. PMID- 18394163 TI - The katG mRNA of Mycobacterium tuberculosis and Mycobacterium smegmatis is processed at its 5' end and is stabilized by both a polypurine sequence and translation initiation. AB - BACKGROUND: In Mycobacterium tuberculosis and in Mycobacterium smegmatis the furA katG loci, encoding the FurA regulatory protein and the KatG catalase-peroxidase, are highly conserved. In M. tuberculosis furA-katG constitute a single operon, whereas in M. smegmatis a single mRNA covering both genes could not be found. In both species, specific 5' ends have been identified: the first one, located upstream of the furA gene, corresponds to transcription initiation from the furA promoter; the second one is the katG mRNA 5' end, located in the terminal part of furA. RESULTS: In this work we demonstrate by in vitro transcription and by RNA polymerase Chromatin immunoprecipitation that no promoter is present in the M. smegmatis region covering the latter 5' end, suggesting that it is produced by specific processing of longer transcripts. Several DNA fragments of M. tuberculosis and M. smegmatis were inserted in a plasmid between the sigA promoter and the lacZ reporter gene, and expression of the reporter gene was measured. A polypurine sequence, located four bp upstream of the katG translation start codon, increased beta-galactosidase activity and stabilized the lacZ transcript. Mutagenesis of this sequence led to destabilization of the mRNA. Analysis of constructs, in which the polypurine sequence of M. smegmatis was followed by an increasing number of katG codons, demonstrated that mRNA stability requires translation of at least 20 amino acids. In order to define the requirements for the 5' processing of the katG transcript, we created several mutations in this region and analyzed the 5' ends of the transcripts: the distance from the polypurine sequence does not seem to influence the processing, neither the sequence around the cutting point. Only mutations which create a double stranded region around the processing site prevented RNA processing. CONCLUSION: This is the first reported case in mycobacteria, in which both a polypurine sequence and translation initiation are shown to contribute to mRNA stability. The furA-katG mRNA is transcribed from the furA promoter and immediately processed; this processing is prevented by a double stranded RNA at the cutting site, suggesting that the endoribonuclease responsible for the cleavage cuts single stranded RNA. PMID- 18394164 TI - Human granulocyte colony stimulating factor (hG-CSF): cloning, overexpression, purification and characterization. AB - BACKGROUND: Biopharmaceutical drugs are mainly recombinant proteins produced by biotechnological tools. The patents of many biopharmaceuticals have expired, and biosimilars are thus currently being developed. Human granulocyte colony stimulating factor (hG-CSF) is a hematopoietic cytokine that acts on cells of the neutrophil lineage causing proliferation and differentiation of committed precursor cells and activation of mature neutrophils. Recombinant hG-CSF has been produced in genetically engineered Escherichia coli (Filgrastim) and successfully used to treat cancer patients suffering from chemotherapy-induced neutropenia. Filgrastim is a 175 amino acid protein, containing an extra N-terminal methionine, which is needed for expression in E. coli. Here we describe a simple and low-cost process that is amenable to scaling-up for the production and purification of homogeneous and active recombinant hG-CSF expressed in E. coli cells. RESULTS: Here we describe cloning of the human granulocyte colony stimulating factor coding DNA sequence, protein expression in E. coli BL21(DE3) host cells in the absence of isopropyl-beta-D-thiogalactopyranoside (IPTG) induction, efficient isolation and solubilization of inclusion bodies by a multi step washing procedure, and a purification protocol using a single cationic exchange column. Characterization of homogeneous rhG-CSF by size exclusion and reverse phase chromatography showed similar yields to the standard. The immunoassay and N-terminal sequencing confirmed the identity of rhG-CSF. The biological activity assay, in vivo, showed an equivalent biological effect (109.4%) to the standard reference rhG-CSF. The homogeneous rhG-CSF protein yield was 3.2 mg of bioactive protein per liter of cell culture. CONCLUSION: The recombinant protein expression in the absence of IPTG induction is advantageous since cost is reduced, and the protein purification protocol using a single chromatographic step should reduce cost even further for large scale production. The physicochemical, immunological and biological analyses showed that this protocol can be useful to develop therapeutic bioproducts. In summary, the combination of different experimental strategies presented here allowed an efficient and cost-effective protocol for rhG-CSF production. These data may be of interest to biopharmaceutical companies interested in developing biosimilars and healthcare community. PMID- 18394165 TI - Longitudinal association of body mass index with lung function: the CARDIA study. AB - BACKGROUND: Lung function at the end of life depends on its peak and subsequent decline. Because obesity is epidemic in young adulthood, we quantified age related changes in lung function relative to body mass index (BMI). METHODS: The Coronary Artery Risk Development in Young Adults (CARDIA) study in 1985-86 (year 0) recruited 5,115 black and white men and women, aged 18-30. Spirometry testing was conducted at years 0, 2, 5 and 10. We estimated 10 year change in FVC, FEV1 and FEV1/FVC according to baseline BMI and change in BMI within birth cohorts with initial average ages 20, 24, and 28 years, controlling for race, sex, smoking, asthma, physical activity, and alcohol consumption. MEASUREMENTS AND MAIN RESULTS: Participants with baseline BMI < 21.3 kg/m2 experienced 10 year increases of 71 ml in FVC and 60 ml in FEV1 and neither measure declined through age 38. In contrast, participants with baseline BMI > or = 26.4 kg/m2 experienced 10 year decreases of 185 ml in FVC and 64 ml in FEV1. FEV1/FVC increased with increasing BMI. Weight gain was also associated with lung function. Those who gained the most weight over 10 years had the largest decrease in FVC, but FVC increased with weight gain in those initially thinnest. In contrast, FEV1 decreased with increasing weight gain in all participants, with maximum decline in obese individuals who gained the most weight during the study. CONCLUSION: Among healthy young adults, increasing BMI in the initially thin participants was associated with increasing then stable lung function through age 38, but there were substantial lung function losses with higher and increasing fatness. These results suggest that the obesity epidemic threatens the lung health of the general population. PMID- 18394166 TI - Extrapulmonary tuberculosis, human immunodeficiency virus, and foreign birth in North Carolina, 1993 - 2006. AB - BACKGROUND: The proportion of extrapulmonary tuberculosis (EPTB) reported in the United States has been gradually increasing. HIV infection and foreign birth are increasingly associated with tuberculosis and understanding their effect on the clinical presentation of tuberculosis is important. METHODS: Case-control study of 6,124 persons with tuberculosis reported to the North Carolina Division of Public health from January 1, 1993 to December 31, 2006. Multivariate logistic regression was used to obtain adjusted odds ratios measuring the associations of foreign birth region and US born race/ethnicity, by HIV status, with EPTB. RESULTS: Among all patients with tuberculosis, 1,366 (22.3%) had EPTB, 563 (9.2%) were HIV co-infected, and 1,299 (21.2%) were foreign born. Among HIV negative patients, EPTB was associated with being foreign born (adjusted ORs 1.36 to 5.09, depending on region of birth) and with being US born, Black/African American (OR 1.84; 95% CI 1.42, 2.39). Among HIV infected patients, EPTB was associated with being US born, Black/African American (OR 2.60; 95% CI 1.83, 3.71) and with foreign birth in the Americas (OR 5.12; 95% CI 2.84, 9.23). CONCLUSION: Foreign born tuberculosis cases were more likely to have EPTB than US born tuberculosis cases, even in the absence of HIV infection. Increasing proportions of foreign born and HIV-attributable tuberculosis cases in the United States will likely result in a sustained burden of EPTB. Further research is needed to explore why the occurrence and type of EPTB differs by region of birth and whether host genetic and/or bacterial variation can explain these differences in EPTB. PMID- 18394167 TI - The Merendino procedure following preoperative imatinib mesylate for locally advanced gastrointestinal stromal tumor of the esophagogastric junction. AB - BACKGROUND: Gastrointestinal stromal tumors (GIST) of the esophagogastric junction might pose a major problem to surgical resection. If locally advanced, extended or multivisceral resection with relevant procedural-specific morbidity and mortality is often necessary. CASE PRESENTATION: We report a case of a patient with a locally advanced GIST of the esophagogastric junction who was treated by transhiatal resection of the lower esophagus and gastric cardia with reconstruction by interposition of segment of the jejunum (Merendino procedure). Prior to resection, downsizing of the tumor had successfully been achieved by treatment with imatinib mesylate for six months. Histological proof of GIST by immunohistochemical expression of c-KIT and/or PDGF alpha Receptor is crucial to allow embarking on this treatment strategy. CONCLUSION: A multimodal approach for an advanced GIST of the esophagogastric junction with preoperative administration of imatinib mesylate could avoid extended resection. The Merendino procedure might be considered as the reconstruction method of choice after resection of GIST at this location. PMID- 18394169 TI - Assessment of the efficacy of antimalarial drugs recommended by the National Malaria Control Programme in Madagascar: up-dated baseline data from randomized and multi-site clinical trials. AB - BACKGROUND: In order to improve the monitoring of the antimalarial drug resistance in Madagascar, a new national network based on eight sentinel sites was set up. In 2006/2007, a multi-site randomized clinical trial was designed to assess the therapeutic efficacy of chloroquine (CQ), sulphadoxine-pyrimethamine (SP), amodiaquine (AQ) and artesunate plus amodiaquine combination (ASAQ), the antimalarial therapies recommended by the National Malaria Control Programme (NMCP). METHODS: Children between six months and 15 years of age, with uncomplicated falciparum malaria, were enrolled. Primary endpoints were the day 14 and day-28 risks of parasitological failure, either unadjusted or adjusted by genotyping. Risks of clinical and parasitological treatment failure after adjustment by genotyping were estimated using Kaplan-Meier survival analysis. Secondary outcomes included fever clearance, parasite clearance, change in haemoglobin levels between Day 0 and the last day of follow-up, and the incidence of adverse events. RESULTS: A total of 1,347 of 1,434 patients (93.9%) completed treatment and follow-up to day 28. All treatment regimens, except for the chloroquine (CQ) treatment group, resulted in clinical cure rates above 97.6% by day-14 and 96.7% by day-28 (adjusted by genotyping). Parasite and fever clearance was more rapid with artesunate plus amodiaquine, but the extent of haematological recovery on day-28 did not differ significantly between the four groups. No severe side-effects were observed during the follow-up period. CONCLUSION: These findings (i) constitute an up-dated baseline data on the efficacy of antimalarial drugs recommended by the NMCP, (ii) show that antimalarial drug resistance remains low in Madagascar, except for CQ, compared to the bordering countries in the Indian Ocean region such as the Comoros Archipelago and (iii) support the current policy of ASAQ as the first-line treatment in uncomplicated falciparum malaria. PMID- 18394168 TI - Cost of antipsychotic polypharmacy in the treatment of schizophrenia. AB - BACKGROUND: This study compared the costs of antipsychotic polypharmacy for patients who initiated on 1 of the 3 most commonly prescribed atypical antipsychotics - olanzapine, quetiapine, or risperidone. METHODS: Data were drawn from a large, prospective, naturalistic, multi-site, nonrandomized study of treatment for schizophrenia in the United States conducted between July 1997 and September 2003. Participants who were initiated on olanzapine (N = 405), quetiapine (N = 115), or risperidone (N = 276) were followed for 1 year post initiation and compared on: (a) average daily cost of the index antipsychotic while on the index antipsychotic, (b) average daily cost of the coprescribed antipsychotics while on the index antipsychotic, (c) average daily cost of the index antipsychotic and the coprescribed antipsychotics while on the index antipsychotic, (d) total annual cost of antipsychotic medications prescribed in the year following initiation on the index antipsychotic, using propensity score adjusted bootstrap resampling method. Average daily antipsychotic costs and total annual antipsychotic costs were also estimated using more recent (2004) antipsychotic drug prices. RESULTS: During the 1 year following initiation on the index antipsychotic, the total average daily cost of the index antipsychotic was higher for quetiapine ($15.33) than olanzapine ($13.90, p < .05) and risperidone ($11.04, p < .01), although the average daily cost of the index antipsychotic was higher for olanzapine ($10.08) than risperidone ($6.74, p < .01) or quetiapine ($6.63, p < .01). Lower total average daily costs were observed in risperidone than olanzapine or quetiapine. Significantly lower average daily cost of concomitant antipsychotic medications for olanzapine ($3.82) compared to quetiapine ($8.70, p < .01) or risperidone-initiated patients ($4.30, p < .01) contributed to the lower average daily cost of all antipsychotic medication for olanzapine-initiated patients. Each dollar spent on the index antipsychotic was accompanied by spending an additional $1.31 on concomitant antipsychotics for quetiapine compared to $0.64 for risperidone and $0.38 for olanzapine-initiated patients. A separate intent-to-treat analysis of the total annual antipsychotic cost found a significantly higher total annual antipsychotic cost for quetiapine initiated patients ($5320) compared to olanzapine ($4536, p < .01) or risperidone ($3813, p < .01). CONCLUSION: Prevalent antipsychotic polypharmacy adds substantial cost to the treatment of schizophrenia. Comparison of medication costs need to address the costs of all antipsychotics. A better understanding of concomitant antipsychotic costs provides a more accurate portrayal of antipsychotic medication costs in the treatment of schizophrenia. PMID- 18394170 TI - Non-cell autonomous impairment of oligodendrocyte differentiation precedes CNS degeneration in the Zitter rat: implications of macrophage/microglial activation in the pathogenesis. AB - BACKGROUND: The zitter (zi/zi) rat, a loss-of-function mutant of the glycosylated transmembrane protein attractin (atrn), exhibits widespread age-dependent spongiform degeneration, hypomyelination, and abnormal metabolism of reactive oxygen species (ROS) in the brain. To date, the mechanisms underlying these phenotypes have remained unclear. RESULTS: Here, we show differentiation defects in zi/zi oligodendrocytes, accompanied by aberrant extension of cell-processes and hypomyelination. Axonal bundles were relatively preserved during postnatal development. With increasing in age, the injured oligodendrocytes in zi/zi rats become pathological, as evidenced by the accumulation of iron in their cell bodies. Immunohistochemical analysis revealed that atrn expression was absent from an oligodendrocyte lineage, including A2B5-positive progenitors and CNPase positive differentiated cells. The number and distribution of Olig2-positive oligodendrocyte progenitors was unchanged in the zi/zi brain. Furthermore, an in vitro differentiation assay of cultured oligodendrocyte progenitors prepared from zi/zi brains revealed their normal competence for proliferation and differentiation into mature oligodendrocytes. Interestingly, we demonstrated the accelerated recruitment of ED1-positive macrophages/microglia to the developing zi/zi brain parenchyma prior to the onset of hypomyelination. Semiquantitative RT PCR analysis revealed a significant up-regulation of CD26 and IL1-beta in the zi/zi brain during this early postnatal stage. CONCLUSION: We demonstrated that the onset of the impairment of oligodendrocyte differentiation occurs in a non cell autonomous manner in zi/zi rats. Hypomyelination of oligodendrocytes was not due to a failure of the intrinsic program of oligodendrocytes, but rather, was caused by extrinsic factors that interrupt oligodendrocyte development. It is likely that macrophage/microglial activation in the zi/zi CNS leads to disturbances in oligodendrocyte differentiation via deleterious extrinsic factors, such as the cytokine IL1-beta or ROS. Atrn might be involved in the activation of brain macrophages/microglia by suppressing excessive migration of monocytes into the CNS, or by accelerating the transformation of brain monocytes into resting microglia. Understanding the pathogenesis of the zi/zi rat may provide novel insights into the developmental interaction betweens macrophages/microglia and cells of an oligodendrocyte lineage. PMID- 18394171 TI - Place of birth, duration of residence, neighborhood immigrant composition and body mass index in New York City. AB - BACKGROUND: Past research has suggested that changes in culture explain the substantial weight gain seen in many immigrant groups with length of residence in the U.S. and across generations of residence in the U.S. However, it has been theorized that those settling in immigrant and co-ethnic neighborhoods may be buffered against this acculturative process and will be more likely to maintain home country dietary and physical activity patterns. To investigate this theory we incorporated measures of neighborhood immigrant composition into analyses of individual's body mass index (BMI) and generation of immigration and duration of residence in the U.S. METHODS: Multilevel analyses were performed using objectively measured height and weight and survey data on diet and physical activity from a sample of 13,011 residents of New York City. Census data were used to calculate the proportion of foreign-born residents and extent of household linguistic isolation in a (1/2) mile radial buffer around the subject's home. RESULTS: Foreign birth was associated with a significantly lower BMI (-1.09 BMI units, P < 0.001). This association was weakest among Asians (-0.66 BMI units, P = 0.08) and strongest among Black-Caribbeans (-1.41 BMI units, P = 0.07). After controlling for individual level variables, neighborhood proportion foreign-born was not associated with BMI, but increasing neighborhood linguistic isolation was inversely associated with BMI among Hispanics (-2.97 BMI units, P = 0.03). Furthermore among Hispanics, the association between foreign birth and BMI was stronger in low linguistic isolation neighborhoods (-1.36 BMI units, P < 0.0001) as compared to in high linguistic isolation levels (-0.42 BMI units, P = 0.79). Increasing duration of residence in the U.S. was significantly associated with higher BMI overall and among Hispanics. CONCLUSION: The analyses suggest that acculturation is associated with weight gain, and that neighborhood characteristics are only associated with BMI among Hispanics. However, we suggest that changes in body size currently interpreted as post-migration effects of acculturation to U.S. norms may in fact reflect changes in norms that are taking place internationally. PMID- 18394172 TI - Molecular analysis reveals heterogeneity of mouse mammary tumors conditionally mutant for Brca1. AB - BACKGROUND: Development of therapies for patients with BRCA1 mutations has been hampered by lack of readily available in vitro and in vivo models. We recently showed that transplantation of transgenic mammary tumors as cell suspensions into naive recipients generates reproducible tumors with remarkable stability of gene expression profile. We examined the expression profiles of original and serially transplanted mammary tumors from Brca1 deficient mice, and tumor derived cell lines to validate their use for preclinical testing and studies of tumor biology. METHODS: Original tumors, serially transplanted and multiple cell lines derived from Brca1 mammary tumors were characterized by morphology, gene and protein expression, and cell surface markers. RESULTS: Gene expression among Brca1 tumors showed more heterogeneity than among previously characterized tumors from MMTV PyMT and -Wnt1 models. Gene expression data segregated Brca1 tumors into 3 distinct types: basal, mixed luminal, and tumors with epithelial-to-mesenchymal transition (EMT). Serial transplantation of individual tumors and multiple cell lines derived from the original tumors recapitulated the molecular characteristics of each tumor of origin. One tumor had distinct features of EMT and gave rise to cell lines that contained a distinct CD44+/CD24-/low population that may correlate with human breast cancer stem cells. CONCLUSION: Although individual tumors expanded by transplantation maintain the genomic profile of the original tumors, the heterogeneity among Brca1 tumors limits the extent of their use for preclinical testing. However, cell lines offer a robust material for understanding tumor biology and response to therapies driven by BRCA1 deficiency. PMID- 18394173 TI - Prevalence of prehypertension and associated cardiovascular risk profiles among prediabetic Omani adults. AB - BACKGROUND: The importance of prediabetes and prehypertension has been demonstrated in several studies especially for primary prevention of CVD. A recent community based cross-sectional study revealed that 40.9 percent Omani adults are prediabetics. This study was undertaken to estimate the prevalence of prehypertension and associated cardiovascular risk profiles in prediabetics. To best of our knowledge, this is the first report on this subject. METHODS: The study included 327 confirmed pre-diabetic Omani adults, who were analysed for their demographic, metabolic and behavioral characteristics. These characteristics were compared between the three different blood pressure groups to determine the CVD risk factors. Univariate and step-wise multiple logistic regression methods were used to carry out the analysis of the data. RESULTS: In this study, the prevalence of prehypertension was 54.1 percent. Males were at higher risk of developing prehypertension as compared to females (OR = 2.30, 95% CI: 1.21, 4.38; P < 0.01). The individuals with higher BMI have two fold more risk of developing prehypertension (OR = 2.25, 95% CI: 1.26, 4.02; P < 0.01). The increased level of OGT enhanced the risk of developing prehypertension (OR = 1.26, 95% CI: 1.06, 1.50; P < 0.01). CONCLUSION: A high prevalence of prehypertension (54.1%) exists in this study population. The major determinants of prehypertension in these prediabetic subjects were male gender, increasing dysglycemia and BMI. Appropriate intervention strategies have been suggested. PMID- 18394176 TI - Bilateral olfaction: two is better than one for navigation. AB - Do animals require bilateral input to track odors? A recent study reveals that fruit fly larvae can localize odor sources using unilateral inputs from a single functional sensory neuron, but that an enhanced signal-to-noise ratio provided by dual inputs is helpful in more challenging environments. PMID- 18394174 TI - ModuleMiner - improved computational detection of cis-regulatory modules: are there different modes of gene regulation in embryonic development and adult tissues? AB - We present ModuleMiner, a novel algorithm for computationally detecting cis regulatory modules (CRMs) in a set of co-expressed genes. ModuleMiner outperforms other methods for CRM detection on benchmark data, and successfully detects CRMs in tissue-specific microarray clusters and in embryonic development gene sets. Interestingly, CRM predictions for differentiated tissues exhibit strong enrichment close to the transcription start site, whereas CRM predictions for embryonic development gene sets are depleted in this region. PMID- 18394177 TI - Recently published papers: Sepsis--guidelines, treatment and novel approaches. AB - The choice of inotropic agent, particularly in catecholamine-resistant septic shock, remains an area of debate. Here we discuss a recent trial examining the use of vasopressin in a carefully controlled trial setting. Yet more data on the use of drotrecogin alfa (activated) in septic shock are described, as are novel but as yet experimental approaches to the treatment of sepsis. Finally, it is important not to forget to read the latest surviving sepsis guidelines. PMID- 18394178 TI - Cardiovascular and metabolic responses to catecholamine and sepsis prognosis: a ubiquitous phenomenon? AB - Many parameters have been associated with sepsis prognosis. In the present issue of Critical Care, Kumar and colleagues demonstrate that a preserved cardiac answer to dobutamine evaluated by radionucleotide measurements was associated with a better prognosis during septic shock. In this context, it is interesting to note that not only is the cardiac response to catecholamine stimulation associated with prognosis, but also the vascular and metabolic responses are associated. The ability of exogenous catecholamine to increase the arterial pressure (dopamine test) or to increase the lactate level is also related to prognosis. According to the ubiquitous character of catecholamine sensitivity, therefore, we should think in terms of cellular ability to respond to catecholamines in defining the concept of physiological reserve. PMID- 18394179 TI - Advances in the genetics of rheumatoid arthritis point to subclassification into distinct disease subsets. AB - In the past few years considerable advances have been made in the genetics of susceptibility to rheumatoid arthritis (RA). For decades the HLA-DRB1 alleles were the only extensively replicated genetic factor, but more genetic risk factors have now been identified that predispose to RA. Interestingly, several of the observed genetic variants conferred risk to anticitrulline-peptide antibody (ACPA)-positive RA and two variants may be restricted to ACPA-negative RA, pointing to the need for subclassification of RA. The current manuscript reviews recently identified genetic factors predisposing to ACPA-positive RA and ACPA negative RA. Additionally, although being scarcely explored, genetic variants affecting the severity of disease course are discussed. PMID- 18394180 TI - Circulating pro-apoptotic mediators in burn septic acute renal failure. AB - The pathogenesis of septic acute kidney injury (AKI) is not well understood. In the present issue of Critical Care, the combined clinical and experimental study from Mariano's group provides new insight into the disease. The study shows that plasma from septic burn patients with acute renal failure initiated pro-apoptotic effects and functional alterations in renal tubular cells and podocytes in vitro that correlated with the degree of proteinuria and renal dysfunction. Pro apoptotic effects were not attributable to antibiotic or uremic toxicity, but were partially attributable to endotoxin. Sepsis and burn had additive effects. Apart from increasing our understanding of the pathogenesis of septic AKI, the study justifies further research on therapeutic interventions in several directions. These include the binding and elimination of the source of endotoxin by selective decontamination of the digestive tract, the blocking of apoptotic pathways, or the extracorporeal removal of circulating toxic mediators using high permeability hemofiltration or coupled plasma filtration and absorption. PMID- 18394181 TI - Towards goal-directed therapy of hepatorenal syndrome: we have the tools but we need the trials. AB - Patients with cirrhosis who develop tense ascites and hepatorenal syndrome have a very high mortality and present a management challenge. Current debate stems from a lack of studies evaluating changes in effective arterial blood volume following paracentesis or targeting fluid replacement with appropriate vascular physiological measures to ensure no paracentesis-related circulatory dysfunction. The study by Umgelter and colleagues addresses a goal-directed approach to fluid management in hepatorenal syndrome and raises several mechanistic questions, the answers to which are likely to improve our understanding of the pathophysiology in hepatorenal syndrome and to guide future management. PMID- 18394182 TI - Optimal vasopressor drug therapy during resuscitation. AB - Optimal vasopressor support during resuscitation should theoretically enhance aortic diastolic and coronary perfusion pressure as well as coronary and cerebral blood flow/oxygen delivery without increasing cellular oxygen demand. Intravenous vasopressor support, using 1 mg doses of epinephrine every 5 minutes in adults or vasopressin 40 IU, is recommended by American Heart Association Advanced Cardiac Life Support Guidelines to maximize oxygen delivery to the heart and brain and increase cellular high energy phosphate levels. Vasopressin offers theoretical advantages over epinephrine in that it does not increase myocardial oxygen demand significantly and its receptors are relatively unaffected by acidosis. However, unlike epinephrine, it is not a myocardial stimulant. Despite these differences in physiologic actions, two large randomized clinical trials yielded virtually identical overall survival to hospital discharge when these agents were compared during inhospital or out-of-hospital resuscitation in Canada and Europe, respectively. More recent clinical and experimental evidence suggests that a combination of vasopressin and epinephrine used during resuscitation can improve hemodynamics and perhaps survival. The verdict on a combination vasopressor strategy may soon come from a large (>2,000 patients) prospective clinical trial that is underway in France to clarify the role of combination vasopressin/epinephrine therapy in out-of-hospital resuscitation. PMID- 18394183 TI - Pulmonary artery occlusion pressure estimation by transesophageal echocardiography: is simpler better? AB - The measurement of pulmonary artery occlusion pressure (PAOP) is important for estimation of left ventricular filling pressure and for distinction between cardiac and non-cardiac etiology of pulmonary edema. Clinical assessment of PAOP, which relies on physical signs of pulmonary congestion, is uncertain. Reliable PAOP measurement can be performed by pulmonary artery catheter, but it is possible also by the use of echocardiography. Several Doppler variables show acceptable correlation with PAOP and can be used for its estimation in cardiac and critically ill patients. Noninvasive PAOP estimation should probably become an integral part of transthoracic and transesophageal echocardiographic evaluation in critically ill patients. However, the limitations of both methods should be taken into consideration, and in specific patients invasive PAOP measurement is still unavoidable, if the exact value of PAOP is needed. PMID- 18394184 TI - Tracheostomy decannulation: marathons and finish lines. AB - Critically ill patients with a tracheostomy who are recovering from respiratory failure eventually require evaluation for airway decannulation. Although expert recommendations guide decisions for managing decannulation, few if any investigative data exist to inform evidence-based care. Consequently, practice variation limits the effectiveness of weaning from tracheostomy. In an investigation reported in this issue of Critical Care, the authors surveyed experienced physicians and respiratory therapists to assess their opinions on managing airway decannulation and identified several clinical factors that they recommend for selecting patients for tracheostomy tube removal. The authors propose that these factors can assist with designing clinical trials of tracheostomy decannulation. Pending completion of such studies, this report underscores the problem of practice variation in managing tracheotomized patients after critical illness. An important implication of the study is that care providers should recognize our knowledge deficit and develop systematic protocols for improving patient care using quality improvement techniques. Such models exist in the literature for adult patients and for children with tracheostomies who are managed by expert teams with requisite knowledge and skills. PMID- 18394185 TI - Knowing who would respond to a recruitment maneuver before actually doing it- this might be a way to go. AB - Using recruitment manoeuvres in acute lung injury remains a controversial issue because no convincing outcome data support their general use, although many physiological studies have demonstrated beneficial effects on lung compliance, end-expiratory lung volume and gas exchange. One of the reasons why physiologically meaningful observations do not translate into clear clinical benefit could be the heterogeneity of the studied patient population. In patients with consolidated lungs and only limited potential for recruitment, manoeuvres might be harmful, whereas in patients with high potential for recruitment they might be helpful. However, when those populations are mixed any signal may be lost because of counteracting effects, depending on how the patient population was mixed. We do not currently have any simple tool that may readily be applied at the bedside to assess the recruitment potential in an individual patient, which would be a sine qua non for identifying a homogeneous population in a recruitment study. Therefore, the method presented by Jacob Koefeld-Nielsen and colleagues in the previous issue of Critical Care provides us with a simple method that could be used at the bedside to assess recruitment potential before the manoeuvre is applied. PMID- 18394186 TI - Myocardial infarction on the ICU: can we do better? AB - Myocardial infarction remains a major cause of death despite contemporary therapeutic strategies. Diagnosis in the intensive care unit is challenging, but is essential to target therapy accurately. In this issue of Critical Care Lim and colleagues present the results of a prospective non-interventional screening study for acute myocardial infarction in patients admitted to the intensive care unit. Myocardial infarction is observed to occur frequently, often without being clinically apparent, with a high associated mortality. Such approaches may facilitate accurate diagnosis of myocardial infarction in this setting, hence opening the way to improved therapy. PMID- 18394187 TI - Inflammation and breast cancer: metalloproteinases as common effectors of inflammation and extracellular matrix breakdown in breast cancer. AB - Two rapidly evolving fields are converging to impact breast cancer: one has identified novel substrates of metalloproteinases that alter immune cell function, and the other has revealed a role for inflammation in human cancers. Evidence now shows that the mechanisms underlying these two fields interact in the context of breast cancer, providing new opportunities to understand this disease and uncover novel therapeutic strategies. The metalloproteinase class of enzymes is well studied in mammary gland development and physiology, but mostly in the context of extracellular matrix modification. Aberrant metalloproteinase expression has also been implicated in breast cancer progression, where these genes act as tumor modifiers. Here, we review how the metalloproteinase axis impacts mammary physiology and tumorigenesis and is associated with inflammatory cell influx in human breast cancer, and evaluate its potential as a regulator of inflammation in the mammary gland. PMID- 18394188 TI - The role of bacteria in lactational mastitis and some considerations of the use of antibiotic treatment. AB - BACKGROUND: The role of bacterial pathogens in lactational mastitis remains unclear. The objective of this study was to compare bacterial species in breast milk of women with mastitis and of healthy breast milk donors and to evaluate the use of antibiotic therapy, the symptoms of mastitis, number of health care contacts, occurrence of breast abscess, damaged nipples and recurrent symptoms in relation to bacterial counts. METHODS: In this descriptive study, breast milk from 192 women with mastitis (referred to as cases) and 466 breast milk donors (referred to as controls) was examined bacteriologically and compared using analytical statistics. Statistical analyses were also carried out to test for relationships between bacteriological content and clinical symptoms as measured on scales, prescription of antibiotics, the number of care contacts, occurrence of breast abscess and recurring symptoms. RESULTS: Five main bacterial species were found in both cases and controls: coagulase negative staphylococci (CNS), viridans streptococci, Staphylococcus aureus (S. aureus), Group B streptococci (GBS) and Enterococcus faecalis. More women with mastitis had S. aureus and GBS in their breast milk than those without symptoms, although 31% of healthy women harboured S. aureus and 10% had GBS. There were no significant correlations between bacterial counts and the symptoms of mastitis as measured on scales. There were no differences in bacterial counts between those prescribed and not prescribed antibiotics or those with and without breast abscess. GBS in breast milk was associated with increased health care contacts (p = 0.02). Women with >/= 10(7) cfu/L CNS or viridans streptococci in their breast milk had increased odds for damaged nipples (p = 0.003). CONCLUSION: Many healthy breastfeeding women have potentially pathogenic bacteria in their breast milk. Increasing bacterial counts did not affect the clinical manifestation of mastitis; thus bacterial counts in breast milk may be of limited value in the decision to treat with antibiotics as results from bacterial culture of breast milk may be difficult to interpret. These results suggest that the division of mastitis into infective or non-infective forms may not be practically feasible. Daily follow-up to measure the subsidence of symptoms can help detect those in need of antibiotics. PMID- 18394189 TI - Valsalva retinopathy in pregnancy: a case report. AB - INTRODUCTION: Valsalva retinopathy is a unilateral or bilateral condition that occurs when increased intra-thoracic or intra-abdominal pressure transmitted to the eye causes a sharp rise in the intra-ocular venous pressure, and rupture of superficial retinal capillaries. The patient often gives a history of a recent strenuous physical act, which could have increased the intra-thoracic pressure. Pregnancy is known to be a risk factor for Valsalva retinopathy. CASE PRESENTATION: A 23-year-old woman in her seventh month of pregnancy presented with a history of decreased vision in her left eye of one-week duration. Examination of the affected eye showed best corrected visual acuity of 20/50, and fundus examination revealed a pre-retinal hemorrhage located in the macula. Based on clinical findings, the diagnosis of Valsalva retinopathy was made. CONCLUSION: Retinal hemorrhages can be generated by Valsalva maneuvers. Pregnancy is a known risk factor for Valsalva retinopathy; however, the diagnosis should be made only after excluding other causes of retinal hemorrhages. It is a self-limited event. We report a case of Valsalva retinopathy complicating normal pregnancy and confirm that, to date, there is no evidence to indicate that there is a risk of recurrence following spontaneous vaginal delivery. PMID- 18394191 TI - An ultrasensitive sorting mechanism for EGF receptor endocytosis. AB - BACKGROUND: The Epidermal Growth Factor (EGF) receptor has been shown to internalize via clathrin-independent endocytosis (CIE) in a ligand concentration dependent manner. From a modeling point of view, this resembles an ultrasensitive response, which is the ability of signaling networks to suppress a response for low input values and to increase to a pre-defined level for inputs exceeding a certain threshold. Several mechanisms to generate this behaviour have been described theoretically, the underlying assumptions of which, however, have not been experimentally demonstrated for the EGF receptor internalization network. RESULTS: Here, we present a mathematical model of receptor sorting into alternative pathways that explains the EGF-concentration dependent response of CIE. The described mechanism involves a saturation effect of the dominant clathrin-dependent endocytosis pathway and implies distinct steady-states into which the system is forced for low vs high EGF stimulations. The model is minimal since no experimentally unjustified reactions or parameter assumptions are imposed. We demonstrate the robustness of the sorting effect for large parameter variations and give an analytic derivation for alternative steady-states that are reached. Further, we describe extensibility of the model to more than two pathways which might play a role in contexts other than receptor internalization. CONCLUSION: Our main result is that a scenario where different endocytosis routes consume the same form of receptor corroborates the observation of a clear-cut, stimulus dependent sorting. This is especially important since a receptor modification discriminating between the pathways has not been found experimentally. The model is not restricted to EGF receptor internalization and might account for ultrasensitivity in other cellular contexts. PMID- 18394190 TI - Membrane transporters and protein traffic networks differentially affecting metal tolerance: a genomic phenotyping study in yeast. AB - BACKGROUND: The cellular mechanisms that underlie metal toxicity and detoxification are rather variegated and incompletely understood. Genomic phenotyping was used to assess the roles played by all nonessential Saccharomyces cerevisiae proteins in modulating cell viability after exposure to cadmium, nickel, and other metals. RESULTS: A number of novel genes and pathways that affect multimetal as well as metal-specific tolerance were discovered. Although the vacuole emerged as a major hot spot for metal detoxification, we also identified a number of pathways that play a more general, less direct role in promoting cell survival under stress conditions (for example, mRNA decay, nucleocytoplasmic transport, and iron acquisition) as well as proteins that are more proximally related to metal damage prevention or repair. Most prominent among the latter are various nutrient transporters previously not associated with metal toxicity. A strikingly differential effect was observed for a large set of deletions, the majority of which centered on the ESCRT (endosomal sorting complexes required for transport) and retromer complexes, which - by affecting transporter downregulation and intracellular protein traffic - cause cadmium sensitivity but nickel resistance. CONCLUSION: The data show that a previously underestimated variety of pathways are involved in cadmium and nickel tolerance in eukaryotic cells. As revealed by comparison with five additional metals, there is a good correlation between the chemical properties and the cellular toxicity signatures of various metals. However, many conserved pathways centered on membrane transporters and protein traffic affect cell viability with a surprisingly high degree of metal specificity. PMID- 18394192 TI - Introduction of medical emergency teams in Australia and New Zealand: a multi centre study. AB - INTRODUCTION: Information about Medical Emergency Teams (METs) in Australia and New Zealand (ANZ) is limited to local studies and a cluster randomised controlled trial (the Medical Emergency Response and Intervention Trial [MERIT]). Thus, we sought to describe the timing of the introduction of METs into ANZ hospitals relative to relevant publications and to assess changes in the incidence and rate of intensive care unit (ICU) admissions due to a ward cardiac arrest (CA) and ICU readmissions. METHODS: We used the Australian and New Zealand Intensive Care Society database to obtain the study data. We related MET introduction to publications about adverse events and MET services. We compared the incidence and rate of readmissions and admitted CAs from wards before and after the introduction of an MET. Finally, we identified hospitals without an MET system which had contributed to the database for at least two years from 2002 to 2005 and measured the incidence of adverse events from the first year of contribution to the second. RESULTS: The MET status was known for 131 of the 172 (76.2%) hospitals that did not participate in the MERIT study. Among these hospitals, 110 (64.1%) had introduced an MET service by 2005. In the 79 hospitals in which the MET commencement date was known, 75% had introduced an MET by May 2002. Of the 110 hospitals in which an MET service was introduced, 24 (21.8%) contributed continuous data in the year before and after the known commencement date. In these hospitals, the mean incidence of CAs admitted to the ICU from the wards changed from 6.33 per year before to 5.04 per year in the year after the MET service began (difference of 1.29 per year, 95% confidence interval [CI] -0.09 to 2.67; P = 0.0244). The incidence of ICU readmissions and the mortality for both ICU-admitted CAs from wards and ICU readmissions did not change. Data were available to calculate the change in ICU admissions due to ward CAs for 16 of 62 (25.8%) hospitals without an MET system. In these hospitals, admissions to the ICU after a ward CA decreased from 5.0 per year in the first year of data contribution to 4.2 per year in the following year (difference of 0.8 per year, 95% CI -0.81 to 3.49; P = 0.3). CONCLUSION: Approximately 60% of hospitals in ANZ with an ICU report having an MET service. Most introduced the MET service early and in association with literature related to adverse events. Although available in only a quarter of hospitals, temporal trends suggest an overall decrease in the incidence of ward CAs admitted to the ICU in MET as well as non-MET hospitals. PMID- 18394193 TI - Radical hysterectomy in the elderly. AB - BACKGROUND: The considerable increase in life expectancy on one hand and an increase in cervical cancer among Iranian patients on the other, brings out the importance of investigating whether radical surgery can be performed safely and effectively on patients above 60 years of age. METHODS: In a study of historical cohort, all 22 patients 60 years and above who have undergone a Wertheim radical hysterectomy for cervical cancer from 1999 to 2005 were compared with 128 matched cases under 60 years of age who had undergone a Wertheim hysterectomy during the same calendar year. All patients were analyzed for preexisting medical comorbidities, length of postoperative stay, morbidity, and postoperative mortality. RESULTS: There was no operative mortality in either group, morbidity (minor, p = 0.91; major, p = 0.89) were statistically not different in the two groups despite the patient's above 60 years having significantly higher comorbidity prior to surgery than the younger cohort (minor, P < 0.05; major, P < 0.05). The mean postoperative hospital stay was significantly longer in the older patients (5 days vs. 3 days, P < 0.001). CONCLUSION: Wertheim Radical hysterectomy is a safe surgical procedure in the selected population of patients 60 years and over. No differences in operative mortality or morbidity were found when compared to a cohort of patient's aged 60 years or younger. PMID- 18394194 TI - A rare association of deformities with diplopodia, aplasia of the tibia and double fibula: a case report. AB - INTRODUCTION: The association of fibular duplication with metatarsal diplopodia is extremely rare with only a few cases reported in the medical literature. CASE PRESENTATION: We present a 4-month-old girl with left tibial agenesis with fibular duplication (mirror foot) and metatarsal diplopodia. CONCLUSION: The case report highlights the need for an understanding of this rare congenital anomaly which may be seen only once in the working lifetime of an orthopaedic surgeon. PMID- 18394195 TI - Future interactions in Particle Toxicology: the role of PFT. PMID- 18394196 TI - Even one star at A level could be "too little, too late" for medical student selection. AB - BACKGROUND: More and more medical school applicants in England and Wales are gaining the maximum grade at A level of AAA, and the UK Government has now agreed to pilot the introduction of a new A* grade. This study assessed the likely utility of additional grades of A* or of A**. METHODS: Statistical analysis of university selection data collected by the Universities and Colleges Admissions Service (UCAS), consisting of data from 1,484,650 applicants to UCAS for the years 2003, 2004 and 2005, of whom 23,628 were medical school applicants, and of these 14,510 were medical school entrants from the UK, aged under 21, and with three or four A level results. The main outcome measure was the number of points scored by applicants in their best three A level subjects. RESULTS: Censored normal distributions showed a good fit to the data using maximum likelihood modelling. If it were the case that A* grades had already been introduced, then at present about 11% of medical school applicants and 18% of entrants would achieve the maximum score of 3 A*s. Projections for the years 2010, 2015 and 2020 suggest that about 26%, 35% and 46% of medical school entrants would have 3 A* grades. CONCLUSION: Although A* grades at A level will help in medical student selection, within a decade, a third of medical students will gain maximum grades. While revising the A level system there is a strong argument, as proposed in the Tomlinson Report, for introducing an A** grade. PMID- 18394198 TI - Vitrectomy combined with glial tissue removal at the optic pit in a patient with optic disc pit maculopathy: a case report. AB - INTRODUCTION: We present a case of a man with optic disc pit maculopathy, whose vision improved after vitrectomy combined with glial tissue removal from the optic pit area, and without the use of photocoagulation. CASE PRESENTATION: A 45 year-old man complained of blurred vision, and ophthalmoscopy revealed a retinal detachment and retinoschisis extending from an optic disc pit through the macula in his left eye. He was diagnosed with optic disc pit maculopathy, and vitrectomy was performed. A posterior vitreous detachment was created, glial tissue at the optic pit was removed, and octafluoropropane (C3F8) was injected as a gas tamponade. The retinal detachment and retinoschisis disappeared after six months, and vision improved to 20/20 without any visual field defects (Goldmann perimetry). A cataractous lens was extracted 2 years after the vitrectomy, and vision has remained 20/20 for 10 years without any recurrence. CONCLUSION: The removal of glial tissue during vitrectomy may be beneficial in patients with optic disc pit maculopathy. PMID- 18394197 TI - A novel method of differential gene expression analysis using multiple cDNA libraries applied to the identification of tumour endothelial genes. AB - BACKGROUND: In this study, differential gene expression analysis using complementary DNA (cDNA) libraries has been improved. Firstly by the introduction of an accurate method of assigning Expressed Sequence Tags (ESTs) to genes and secondly, by using a novel likelihood ratio statistical scoring of differential gene expression between two pools of cDNA libraries. These methods were applied to the latest available cell line and bulk tissue cDNA libraries in a two-step screen to predict novel tumour endothelial markers. Initially, endothelial cell lines were in silico subtracted from non-endothelial cell lines to identify endothelial genes. Subsequently, a second bulk tumour versus normal tissue subtraction was employed to predict tumour endothelial markers. RESULTS: From an endothelial cDNA library analysis, 431 genes were significantly up regulated in endothelial cells with a False Discovery Rate adjusted q-value of 0.01 or less and 104 of these were expressed only in endothelial cells. Combining the cDNA library data with the latest Serial Analysis of Gene Expression (SAGE) library data derived a complete list of 459 genes preferentially expressed in endothelium. 27 genes were predicted tumour endothelial markers in multiple tissues based on the second bulk tissue screen. CONCLUSION: This approach represents a significant advance on earlier work in its ability to accurately assign an EST to a gene, statistically measure differential expression between two pools of cDNA libraries and predict putative tumour endothelial markers before entering the laboratory. These methods are of value and available http://www.compbio.ox.ac.uk/data/diffex.html to researchers that are interested in the analysis of transcriptomic data. PMID- 18394199 TI - Hypertrophic osteoarthropathy as the cause of a super scan of the bone in a patient with prostate cancer: a case report. AB - INTRODUCTION: Prostate cancer is known to have a tendency to metastasize to bone. Skeletal scintigraphy can be used to show multiple lesions. Diffuse metastasis, which is not infrequent in prostate cancer, can also be suspected on the basis of a 'super scan'. However, this phenomenon in nuclear medicine has several other causes that need to be considered. CASE PRESENTATION: A patient with a history of prostate cancer presented with pleural fluid, peripheral edema and bone pain. A super scan of the bone was found which suggested diffuse skeletal metastasis of the prostate cancer but the patient also had a prostate specific antigen level which was not compatible with this diagnosis. Further investigations revealed the paraneoplastic phenomenon of hypertrophic osteoarthropathy, related to an incurable carcinoma of the lung, to be the cause of the super scan. CONCLUSION: A super scan is characterized by a high bone to soft tissue ratio on skeletal scintigraphy, with a uniform symmetrical increase in bone uptake and diminished to absent renal visualization ('absent kidney sign'). It can be seen in a variety of diseases in which there is a diffusely increased bone turnover. Diffuse skeletal metastasis of a well-differentiated prostate carcinoma is unlikely to be the cause of a super scan when the prostate specific antigen level is not elevated. This is the first report of a super scan due to pulmonary hypertrophic osteoarthropathy which can be seen in lung carcinoma and other pulmonary diseases. PMID- 18394200 TI - Interventions designed to improve the quality and efficiency of medication use in managed care: a critical review of the literature - 2001-2007. AB - BACKGROUND: Managed care organizations use a variety of strategies to reduce the cost and improve the quality of medication use. The effectiveness of such policies is not well understood. The objective of this research was to update a previous systematic review of interventions, published between 1966 and 2001, to improve the quality and efficiency of medication use in the US managed care setting. METHODS: We searched MEDLINE and EMBASE for publications from July 2001 to January 2007 describing interventions targeting drug use conducted in the US managed care setting. We categorized studies by intervention type and adequacy of research design using commonly accepted criteria. We summarized the outcomes of well-controlled strategies and documented the significance and magnitude of effects for key study outcomes. RESULTS: We identified 164 papers published during the six-year period. Predominant strategies were: educational interventions (n = 20, including dissemination of educational materials, and group or one-to-one educational outreach); monitoring and feedback (n = 22, including audit/feedback and computerized monitoring); formulary interventions (n = 66, including tiered formulary and patient copayment); collaborative care involving pharmacists (n = 15); and disease management with pharmacotherapy as a primary focus (n = 41, including care for depression, asthma, and peptic ulcer disease). Overall, 51 studies met minimum criteria for methodological adequacy. Effective interventions included one-to-one academic detailing, computerized alerts and reminders, pharmacist-led collaborative care, and multifaceted disease management. Further, changes in formulary tier-design and related increases in copayments were associated with reductions in medication use and increased out-of pocket spending by patients. The dissemination of educational materials alone had little or no impact, while the impact of group education was inconclusive. CONCLUSION: There is good evidence for the effectiveness of several strategies in changing drug use in the managed care environment. However, little is known about the cost-effectiveness of these interventions. Computerized alerts showed promise in improving short-term outcomes but little is known about longer-term outcomes. Few well-designed, published studies have assessed the potential negative clinical effects of formulary-related interventions despite their widespread use. However, some evidence suggests increases in cost sharing reduce access to essential medicines for chronic illness. PMID- 18394202 TI - Relationship between cognitive appraisals of symptoms and negative mood for subtypes of irritable bowel syndrome. AB - BACKGROUND: The onset and course of irritable bowel syndrome (IBS) are strongly influenced by psychological factors, and treatment often includes cognitive behavioral therapy. We conducted a study of the relationships between cognitive appraisal of IBS symptoms and negative mood for the subtypes of IBS. METHOD: The participants were 1087 college students who completed a set of questionnaires that included the Rome II Modular Questionnaire, Self-reported IBS Questionnaire, Cognitive Appraisal Rating Scale, and the Hospital Anxiety and Depression Scale. RESULTS: The participants included 206 individuals with IBS; 61 had diarrhea predominant IBS (IBSD) and 45 had constipation-predominant IBS (IBSC). The overall IBS group scored higher on anxiety and depression than the control group. The IBSD and IBSC groups each had significantly higher scores for anxiety but did not significantly differ from the control group in scores for depression. There were no significant differences between the IBSD and IBSC groups in their cognitive appraisal of IBS symptoms. For the IBSD group, anxiety was significantly, positively correlated with commitment, effect, and threat, and depression was significantly, negatively correlated with controllability. In contrast, there were no significant correlations between mood and cognitive appraisal for the IBSC group. Multiple regression analyses with abdominal symptoms as dependent variables and cognitive appraisals as independent variables showed that for the IBSD group, abdominal pain was significantly, positively correlated with commitment, and abdominal discomfort was significantly, positively correlated with appraisal of effect and threat. For the IBSC group, abdominal pain and hard stool were significantly, positively correlated with commitment, and abdominal discomfort was significantly, positively correlated with appraisal of effect and threat. CONCLUSION: IBS patients as a general group report high levels of anxiety and depression. However, IBSD and IBSC were both associated only with high anxiety, but not depression, when compared to the non IBS control group. For the IBSD group, anxiety was associated with cognitive appraisals, but this association was not found for the IBSC group. These groups did not differ in their associated cognitive appraisals, and are similar in terms of the positive relationship between abdominal pain and discomfort and the cognitive appraisals of coping. PMID- 18394201 TI - The National Cancer Institute Thyroid fine needle aspiration state of the science conference: a summation. PMID- 18394203 TI - Vasodilative effects of prostaglandin E1 derivate on arteries of nerve roots in a canine model of a chronically compressed cauda equina. AB - BACKGROUND: Reduction of blood flow is important in the induction of neurogenic intermittent claudication (NIC) in lumbar spinal canal stenosis. PGE1 improves the mean walking distance in patients with NIC type cauda equina compression. PGE1 derivate might be effective in dilating blood vessels and improving blood flow in nerve roots with chronically compressed cauda equina. The aim of this study was to assess whether PGE1 derivate has vasodilatory effects on both arteries and veins in a canine model of chronic cauda equina compression. METHODS: Fourteen dogs were used in this study. A plastic balloon inflated to 10 mmHg was placed under the lamina of the 7th lumbar vertebra for 1 week. OP-1206 cyclodextrin clathrate (OP-1206-CD: prostaglandin E1 derivate) was administered orally. The blood vessels of the second or third sacral nerve root were identified using a specially designed surgical microscope equipped with a video camera. The diameter of the blood vessels was measured on video-recordings every 15 minutes until 90 minutes after the administration of the PGE1 derivate. RESULTS: We observed seven arteries and seven veins. The diameter and blood flow of the arteries was significantly increased compared with the veins at both 60 and 75 minutes after administration of the PGE1 derivate (p < 0.05). Blood flow velocity did not change over 90 minutes in either the arteries or veins. DISCUSSION: The PGE1 derivate improved blood flow in the arteries but did not induce blood stasis in the veins. Our results suggest that the PGE1 derivate might be a potential therapeutic agent, as it improved blood flow in the nerve roots in a canine model of chronic cauda equina compression. PMID- 18394204 TI - Not all aggregate-level correlations approach unity: a reply to William Lichten. PMID- 18394205 TI - Evaluation of adenoidal obstruction in children: clinical symptoms compared with roentgenographic assessment. AB - BACKGROUND: Obstructive adenoid enlargement is commonly implicated as the major cause of chronic nasal obstruction in children. Although clinical assessment is considered essential, there is little consensus over its reliability. This study was conducted to determine the correlation between graded symptomatology assessment and roentgenographic assessment of adenoidal obstruction. METHOD: Symptoms assessed included snoring, mouth-breathing and obstructive breathing during sleep. Each symptom was rated on a four-point scale (absent = zero, mild = one, moderate = two and severe = three). We summed the ratings for each child to obtain the symptomatology score. We used an adenoidal-nasopharyngeal ratio parameter to classify roentgenographic assessment into minimal, moderate or marked obstruction. RESULTS: Sixty-four children, 42 boys and 22 girls, aged one to 12 years were enrolled. The clinical symptomatology scores correlated significantly with the roentgenographic ratings of nasopharyngeal airway obstruction (r = 0.419; p = 0.001). The correlation was significant at roentgenographic ratings of minimal obstruction (p < 0.05) and gross obstruction (p < 0.001). Both the symptomatology score and the roentgenographic rating showed significant correlations with patient age (r = -0.657, p < 0.01 and r = -0.340, p < 0.01, respectively). CONCLUSION: Clinical rating of adenoidal symptoms in children provides a reasonably reliable assessment of the presence and severity of nasopharyngeal airway obstruction. This technique of assessment is easy to use and is particularly valid when obstruction is either minimal or gross. PMID- 18394206 TI - Modelling and prediction of weekly incidence of influenza A specimens in England and Wales. AB - We propose a rather simple model, which fits well the weekly human influenza incidence data from England and Wales. A standard way to analyse seasonally varying time-series is to decompose them into different components. The residuals obtained after eliminating these components often do not reveal time dependency and are normally distributed. We suggest that conclusions should not be drawn only on the basis of residuals and that one should consider the analysis of squared residuals. We show that squared residuals can reveal the presence of the remaining seasonal variation, which is not exhibited by the analysis of residuals, and that the modelling of such seasonal variations undoubtedly improves model fit. PMID- 18394207 TI - Potential role of pregabalin in the treatment of lithium-induced tremor: a case report. PMID- 18394208 TI - Endoscopic approach to juvenile nasopharyngeal angiofibroma: our experience at a tertiary care centre. AB - OBJECTIVE: To evaluate the efficacy of endoscopic exposure and excision for juvenile nasopharyngeal angiofibroma. DESIGN: Prospective analysis of the outcomes of endoscopic removal of the angiofibroma at a tertiary care centre, using endoscopic and radiological parameters. SUBJECTS: Twenty-eight consecutive patients with angiofibroma were included. These patients underwent endonasal endoscopic excision of the lesion, following pre-operative computed tomography and digital subtraction angiography. Post-operative endoscopy and radiology were performed at six weeks to detect any residual lesion, and thereafter at six monthly intervals to detect recurrence. RESULTS: Twenty-one patients (75 per cent) were new cases and seven patients (25 per cent) had recurrent tumour. Pre operative embolisation was performed in 21 patients (75 per cent). An endoscopic endonasal approach was used to access the pterygopalatine fossa. The average blood loss was 228 ml. Residual lesion was noted in one patient (initially with recurrent stage II C pathology). The remaining 27 patients (initially with stage I, II A and II B pathology) were free of residual pathology or recurrence after a minimum follow-up period of 12 months. CONCLUSION: Endoscopic exposure and excision of angiofibroma was found to be effective. PMID- 18394209 TI - Ecological and biological factors involved in the transmission of Echinococcus multilocularis in the French Ardennes. AB - In order to identify the respective importance of the ecological and biological factors involved in the transmission of Echinococcus multilocularis, we estimated grassland vole intermediate host (Microtus sp. and Arvicola terrestris) population densities, in relation to the diet of the definitive host (red fox, Vulpes vulpes) and with the prevalence of E. multilocularis in the fox population. The study was conducted in the Ardennes, north-eastern France, which is an area with a high incidence of alveolar echinococcosis. Surface index methods showed that Microtus was the most abundant intermediate host in the area. Furthermore, Microtus was present in one-third of the 144 faeces and 98 stomach content samples examined and represented more than two-thirds of the rodent occurrences. Red fox predation on Microtus was significantly correlated with Microtus relative abundance. In contrast, the relative abundance of A. terrestris was very low. This species, as well as Clethrionomys glareolus and Apodemus sp., was little consumed. E. multilocularis prevalence in foxes was determined from carcasses and reached 53% (95% confidence interval 45-61%). Intensity of infection varied from 2 to 73,380 worms per fox, with 72% of the sampled worm burden harboured by 8% of the sampled foxes. The selected explanatory variables (sex, year, age class, health and nutritional condition, and season) failed to predict prevalence rate and worm burden. The high prevalence rate in foxes indicates the possibility of intense E. multilocularis transmission, apart from periods, or in landscapes, favourable to large population outbreaks of grassland rodents. PMID- 18394210 TI - Impact of treatment with praziquantel, silymarin and/or beta-glucan on pathophysiological markers of liver damage and fibrosis in mice infected with Mesocestoides vogae (Cestoda) tetrathyridia. AB - Mesocestoides vogae tetrathyridia infection in mice causes hepatocyte injury, hepatic granulomatous inflammmation, liver fibrosis and chronic peritonitis manifested with portal hypertension. To reduce the detrimental effect of parasites on the host liver, the effect of the anthelmintic drug praziquantel (PZQ) in combination with natural products silymarin (an antioxidant) and beta glucan (an immunomodulator) was investigated. The therapeutic effect of drugs was assessed by means of aminotransferase (alanine aminotransferase (ALT) and aspartate aminotransferase (AST)) activities, content of albumin, total proteins and hyaluronic acid (HA) in sera of ICR mice infected with M. vogae larvae. Animals were treated with PZQ suspended in oil emulsion (Group 1), PZQ combined with silymarin incorporated into lipid microspheres (LMS) (Group 2), PZQ combined with beta-glucan incorporated into liposomes (LG) (Group 3), PZQ co-administered with LMS and LG (Group 4). Untreated animals (Group 5) served as the control. Treatment of animals started at the early chronic phase of infection (day 14 p.i.) and lasted 10 days; serum samples were collected on days 0, 7, 14, 25, 28, 31, 35 and 45 p.i. ALT and AST activities were significantly (P < 0.05) decreased in Groups 2, 3 and 4. HA content was significantly (P < 0.05 and 0.01) lower in Groups 2 and 4. Albumin levels were decreased in Groups 2 and 4, total protein concentration decreased in Groups 1 and 3 (P < 0.05 and 0.01). These results showed that combined treatment of PZQ with silymarin and/or beta-glucan was able to ameliorate or suppress fibrogenesis in the liver, protect liver cells from oxidative damage and, possibly, stimulate regeneration of the parenchyma. PMID- 18394211 TI - Serological and molecular studies on Dirofilaria immitis in dogs from Turkey. AB - We estimated the prevalence of Dirofilaria immitis infection in domestic dogs in five Turkish provinces - Sakarya, Kocaeli, Ankara, Elazig and Mersin - using a commercial ELISA kit for detecting circulating antigen and a PCR test for detecting circulating microfilarial DNA. A total of 211 whole-blood and serum samples were collected from dogs of various breeds, ages and life status (owned or stray). Sample population characteristics were recorded and examined for differences in prevalence. Additionally, we collected 15 blood samples from cats (14 owned and 1 stray) from Ankara province and used PCR to detect D. immitis infection. Twenty-seven (12.8%) of 211 dog samples were positive for D. immitis antigen by ELISA. No differences in prevalence were observed by sex (female: 14.4%; male: 10.7%; P>0.05). The prevalence of D. immitis infection varied with age: 11.8% in younger dogs (0.5-2 years) and 17.5% in older dogs (3-5 years). Prevalence between stray dogs (15.2%) and owned dogs (9.3%) did not differ (P>0.05). Prevalence rates were highest in Kocaeli province (18.3%), followed by Ankara (14.8%), Sakarya (12.3%) and Mersin (10.5%) provinces. Prevalence in Elazig province was 0%. No dogs or cats had microfilarial DNA detectable by PCR. PMID- 18394212 TI - The effect of Anonchocephalus chilensis Riggenbach (Eucestoda: Bothriocephalidea) on infracommunity patterns in Genypterus maculatus Tschudi (Osteichthyes: Ophidiidae). AB - The use of parasite body size (i.e. body mass) is a promising proxy to improve the study of patterns in parasite infracommunities, which are usually analysed using only numerical descriptors. This study deals with the importance and effect of the presence of a large endoparasite species, the cestode Anonchocephalus chilensis in a marine fish species, Genypterus maculatus, on the structure of the parasite infracommunities. Numerical and volumetric measures of aggregated properties of parasite infracommunities were compared and their correlation examined. The highly dominant presence of A. chilensis by volume causes a dramatic change in the patterns observed, including a smaller total volume of the remaining species when this volumetrically dominant species is present. However, C-scores and V-ratios, both indices based on null models of species occurrence, do not support the idea of communities structured by interspecific competition. Analyses reveal that numerical and volumetric community descriptors are complementary ways to search for patterns and to reveal processes within these systems. PMID- 18394213 TI - Effect of the dietary fat quality on insulin sensitivity. AB - Recent evidence shows that specific fatty acids affect cell metabolism, modifying the balance between fatty acid oxidation and lipogenesis. These effects may have important implications in addressing the present epidemic of nutrition-related chronic disease. Intake of dietary saturated and n-6 PUFA have increased while n 3 fatty acid intake has decreased. Obesity, type 2 diabetes and insulin resistance are highly prevalent, and both are strongly related to disorders of lipid metabolism characterized by an increased plasma and intracellular fatty acid availability. Thus, it has been hypothesized that change in the quality of dietary fat supply is able to modify the degree of insulin sensitivity. Animal studies provide support for this notion. However, there is limited human data either from normal or diabetic subjects. This review aims to analyse human studies that address this question. To this purpose, the experimental design, dietary compliance, insulin-sensitivity method used and confounding variables are discussed in order to identify the role of dietary fat quality as a risk factor for insulin resistance. Most studies (twelve of fifteen) found no effect relating to fat quality on insulin sensitivity. However, multiple study design flaws limit the validity of this conclusion. In contrast, one of the better designed studies found that consumption of a high-saturated-fat diet decreased insulin sensitivity in comparison to a high-monounsaturated-fat diet. We conclude that the role of dietary fat quality on insulin sensitivity in human subjects should be further studied, using experimental designs that address the limitations of existing data sets. PMID- 18394214 TI - Dietary flavonoid intake and cardiovascular mortality. PMID- 18394215 TI - Estimates of daily net endogenous acid production in the elderly UK population: analysis of the National Diet and Nutrition Survey (NDNS) of British adults aged 65 years and over. AB - Dietary intake has been shown to influence acid-base balance in human subjects under tightly controlled conditions. However, the net effect of food groups on alkali/acid loading in population groups is unclear. The aims of the present study were to: (1) quantify estimates of daily net endogenous acid production (NEAP) (mEq/d) in a representative group of British elderly aged 65 years and older; (2) compare and characterise NEAP by specific nutrients and food groups likely to influence dietary acid loading; (3) determine whether geographical location influenced NEAP. The National Diet and Nutrition Survey dataset, consisting of a 4 d weighed record and anthropometric data, was used to estimate dietary acidity. Dietary under-reporters were excluded by analysing only subjects with energy intakes >/= 1.2 x BMR. NEAP was estimated as the dietary potential renal acid load+organic acid excretion, the latter as a multiple of estimated body surface area. NEAP was lower in women compared with men (P < 0.001), and lower than values reported in a Swedish elderly cohort. Lower dietary acidity was significantly associated with higher consumption of fruit and potatoes and lower consumption of meat, bread and eggs (P < 0.02 to P < 0.001). Lower intakes of fish and cheese were associated with lower NEAP in men only (P < 0.01 to P < 0.001). There were regional differences for NEAP, with higher intakes in Scotland/Northern regions compared with Central/South-Western and London/South Eastern regions (P = 0.01). These data provide an insight into the acid generating potential of the diet in the British elderly population, which may have important consequences in this vulnerable group. PMID- 18394216 TI - Rapeseed protein inhibits the initiation of insulin resistance by a high saturated fat, high-sucrose diet in rats. AB - In contrast to the quality of carbohydrates and lipids, little is known on the influence of the type of dietary protein on the development of the metabolic or insulin resistance syndrome. Cysteine intake has been recently documented to impact insulin sensitivity. The aim of this study was to determine whether rapeseed protein, an emergent cysteine-rich protein, could inhibit the onset of the metabolic syndrome. For 9 weeks, rats were fed a diet rich in saturated fats and sucrose, which also included 20 % protein either as milk protein ('Induction' diet I) or rapeseed protein (diet R). A third, control group received an isoenergetic diet containing milk protein but polyunsaturated fats and starch ('Prudent' diet P). Plasma glucose, insulin, TAG and cholesterol, and blood pressure were monitored during the study, glucose tolerance was tested at week 7 and body composition determined at week 9. Plasma glucose, insulin and TAG increased during the experiment and, at week 9, plasma insulin was significantly 34 % lower in the R group and 56 % lower in P group as compared with the I group. The insulin peak after the glucose load was significantly 28-30 % lower in R and P than in I and the insulin sensitivity index was significantly higher in R than in I. Unexpectedly, peripheral fat deposition was slightly higher in R than in I. In this model, substituting rapeseed protein for milk protein had preventive effects on the early onset of insulin resistance, similar to those achieved by manipulating the types of dietary fat and carbohydrates. PMID- 18394217 TI - The effect of feeding frequency on insulin and ghrelin responses in human subjects. AB - Recent work shows that increased meal frequency reduces ghrelin responses in sheep. Human research suggests there is an interaction between insulin and ghrelin. The effect of meal frequency on this interaction is unknown. Therefore, we investigated the effect of feeding frequency on insulin and ghrelin responses in human subjects. Five healthy male volunteers were recruited from the general population: age 24 (SEM 2)years, body mass 75.7 (SEM 3.2) kg and BMI 23.8 (SEM 0.8) kg/m(2). Volunteers underwent three 8-h feeding regimens: fasting (FAST); low-frequency(two) meal ingestion (LOFREQ(MEAL)); high-frequency (twelve) meal ingestion (HIFREQ(MEAL)). Meals were equi-energetic within trials,consisting of 64% carbohydrate, 23% fat and 13% protein. Total energy intake was equal between feeding trials. Total area under the curve for serum insulin and plasma ghrelin responses did not differ between trials (P>0.05), although the hormonal response patterns to the two meal feeding regimens were different. An inverse relationship was found between serum insulin and plasma ghrelin during the FAST andLOFREQ(MEAL) trials (P<0.05); and, in the postprandial period, there was a time delay between insulin responses and successive ghrelin responses.This relationship was not observed during the HIFREQ(MEAL) trial (P>0.05). This study provides further evidence that the postprandial fall in ghrelin might be due, at least partially, to the rise in insulin and that high-frequency feeding may disrupt this relationship. PMID- 18394219 TI - Dietary supplement use and mortality in a cohort of Swedish men--comments by Boucher. PMID- 18394220 TI - A comparison of the effects of kaempferol and quercetin on cytokine-induced pro inflammatory status of cultured human endothelial cells. AB - We investigated the effects of the flavonols kaempferol and quercetin on the expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), endothelial cell selectin (E-selectin), inducible NO synthase (iNOS) and cyclo-oxygenase-2 (COX-2), and on the activation of the signalling molecules NF-kappaB and activator protein-1 (AP-1), induced by a cytokine mixture in cultured human umbilical vein endothelial cells. Inhibition of reactive oxygen and nitrogen species generation did not differ among both flavonols at 1 micromol/l but was significantly stronger for kaempferol at 5-50 micromol/l. Supplementation with increasing concentrations of kaempferol substantially attenuated the increase induced by the cytokine mixture in VCAM-1 (10-50 micromol/l), ICAM-1 (50 micromol/l) and E-selectin (5-50 micromol/l) expression. A significantly inhibitory effect of quercetin on VCAM-1 (10-50 micromol/l), ICAM-1 (50 micromol/l) and E-selectin (50 micromol/l) expression was also observed. Expression of adhesion molecules was always more strongly inhibited in kaempferol-treated than in quercetin-treated cells. The inhibitory effect on iNOS and COX-2 protein level was stronger for quercetin at 5-50 micromol/l. The effect of kaempferol on NF-kappaB and AP-1 binding activity was weaker at high concentrations (50 micromol/l) as compared with quercetin. The present study indicates that differences exist in the modulation of pro inflammatory genes and in the blockade of NF-kappaB and AP-1 by kaempferol and quercetin. The minor structural differences between both flavonols determine differences in their anti-inflammatory properties and in their efficiency in inhibiting signalling molecules. PMID- 18394221 TI - Host specificity is linked to intraspecific variability in the genus Lamellodiscus (Monogenea). AB - SUMMARYWe investigated whether host specificity is linked to variability within species of Lamellodiscus monogeneans, which are gill ectoparasites of the Sparidae. We sampled fish parasites in the northeastern part of the Mediterranean Sea: 4 specialist species, using 1 single host species, and 3 generalist species, using 2 distinct host species. Intraspecific variability was assessed from 2 different datasets. Morphometric variability of the attachment organ, called the haptor, was estimated first from measurements of several sclerified haptoral parts on 102 individuals. Genetic variability was calculated based on comparisons of sequences derived from the first internal transcribed spacer (ITS-1) of nuclear ribosomal DNA of 62 individuals. Morphometric variances in the specialist versus generalist species were compared via principal component analysis and F tests, and uncorrected genetic distances (p-distances) were estimated within each species. We showed that the inter-individual variance of morphometric characters, as well as p-distances, are clearly greater within generalist species than specialist ones. These findings suggest that a relative increase in morphological and molecular variability enhances the possibility to colonize new host species in monogeneans, and supports the hypothesis that intraspecific variability could be a potential determinant of host specificity. PMID- 18394222 TI - The shared circuits model (SCM): how control, mirroring, and simulation can enable imitation, deliberation, and mindreading. AB - Imitation, deliberation, and mindreading are characteristically human sociocognitive skills. Research on imitation and its role in social cognition is flourishing across various disciplines. Imitation is surveyed in this target article under headings of behavior, subpersonal mechanisms, and functions of imitation. A model is then advanced within which many of the developments surveyed can be located and explained. The shared circuits model (SCM) explains how imitation, deliberation, and mindreading can be enabled by subpersonal mechanisms of control, mirroring, and simulation. It is cast at a middle, functional level of description, that is, between the level of neural implementation and the level of conscious perceptions and intentional actions. The SCM connects shared informational dynamics for perception and action with shared informational dynamics for self and other, while also showing how the action/perception, self/other, and actual/possible distinctions can be overlaid on these shared informational dynamics. It avoids the common conception of perception and action as separate and peripheral to central cognition. Rather, it contributes to the situated cognition movement by showing how mechanisms for perceiving action can be built on those for active perception.;>;>The SCM is developed heuristically, in five layers that can be combined in various ways to frame specific ontogenetic or phylogenetic hypotheses. The starting point is dynamic online motor control, whereby an organism is closely attuned to its embedding environment through sensorimotor feedback. Onto this are layered functions of prediction and simulation of feedback, mirroring, simulation of mirroring, monitored inhibition of motor output, and monitored simulation of input. Finally, monitored simulation of input specifying possible actions plus inhibited mirroring of such possible actions can generate information about the possible as opposed to actual instrumental actions of others, and the possible causes and effects of such possible actions, thereby enabling strategic social deliberation. Multiple instances of such shared circuits structures could be linked into a network permitting decomposition and recombination of elements, enabling flexible control, imitative learning, understanding of other agents, and instrumental and strategic deliberation. While more advanced forms of social cognition, which require tracking multiple others and their multiple possible actions, may depend on interpretative theorizing or language, the SCM shows how layered mechanisms of control, mirroring, and simulation can enable distinctively human cognitive capacities for imitation, deliberation, and mindreading. PMID- 18394244 TI - A study of the science of taste: on the origins and influence of the core ideas. AB - Our understanding of the sense of taste is largely based on research designed and interpreted in terms of the traditional four "basic" tastes: sweet, sour, salty, and bitter, and now a few more. This concept of basic tastes has no rational definition to test, and thus it has not been tested. As a demonstration, a preliminary attempt to test one common but arbitrary psychophysical definition of basic tastes is included in this article; that the basic tastes are unique in being able to account for other tastes. This definition was falsified in that other stimuli do about as well as the basic words and stimuli. To the extent that this finding might show analogies with other studies of receptor, neural, and psychophysical phenomena, the validity of the century-long literature of the science of taste based on a few "basics" is called into question. The possible origins, meaning, and influence of this concept are discussed. Tests of the model with control studies are suggested in all areas of taste related to basic tastes. As a stronger alternative to the basic tradition, the advantages of the across fiber pattern model are discussed; it is based on a rational data-based hypothesis, and has survived attempts at falsification. Such "population coding" has found broad acceptance in many neural systems. PMID- 18394264 TI - Spatial and temporal dynamics of dengue fever in Peru: 1994-2006. AB - SUMMARYThe weekly number of dengue cases in Peru, South America, stratified by province for the period 1994-2006 were analysed in conjunction with associated demographic, geographic and climatological data. Estimates of the reproduction number, moderately correlated with population size (Spearman rho=0.28, P=0.03), had a median of 1.76 (IQR 0.83-4.46). The distributions of dengue attack rates and epidemic durations follow power-law (Pareto) distributions (coefficient of determination >85%, P<0.004). Spatial heterogeneity of attack rates was highest in coastal areas followed by mountain and jungle areas. Our findings suggest a hierarchy of transmission events during the large 2000-2001 epidemic from large to small population areas when serotypes DEN-3 and DEN-4 were first identified (Spearman rho=-0.43, P=0.03). The need for spatial and temporal dengue epidemic data with a high degree of resolution not only increases our understanding of the dynamics of dengue but will also generate new hypotheses and provide a platform for testing innovative control policies. PMID- 18394265 TI - Quantifying the effect of exenatide and insulin glargine on postprandial glucose excursions in patients with type 2 diabetes. AB - In this report, we quantify the effects of exenatide and glargine on the relative contributions of fasting and postprandial glucose (PPG) excursion to overall hyperglycemia based on self-monitored blood glucose. After 26 weeks of treatment, insulin glargine reduced fasting glucose to a greater extent than exenatide without significant effect on PPG excursion. The principal effect of exenatide on hyperglycemia was mitigating the rise in PPG with moderate improvement on fasting glucose. These findings may be limited by the fact that glucose measurements were collected through self-monitoring with six time points measured during the daytime, the meals were not standardized and the exact time for glucose measurements was unknown. PMID- 18394266 TI - Comparison of the therapeutic effects of epoetin zeta and epoetin alpha in the correction of renal anaemia. AB - OBJECTIVE: To assess the therapeutic equivalence of epoetin zeta and epoetin alpha for correction of haemoglobin (Hb) concentration in patients with anaemia and chronic kidney disease (CKD) stage 5 maintained on haemodialysis. STUDY DESIGN: In total, 609 patients with CKD and anaemia (Hb < 9 g/dL) were randomly assigned to receive either epoetin zeta or epoetin alpha intravenously, one to three times per week for 24 weeks. Dosing was titrated individually to achieve a stable, target Hb concentration of 11-12 g/dL. Primary endpoints were the mean weekly dose of epoetin per kilogram of body weight and mean Hb concentration during the last 4 weeks of treatment. Safety endpoints were the occurrence of anti-erythropoietin antibodies, ratings of tolerability and adverse events (AEs). RESULTS: Mean (+/- standard deviation [SD]) Hb concentration over the last 4 weeks of treatment was 11.61 +/- 1.27 g/dL for patients receiving epoetin zeta, compared with 11.63 +/- 1.37 g/dL for patients receiving epoetin alpha (95% confidence interval [CI]: -0.25 to 0.20 g/dL). Mean (+/- SD) epoetin zeta weekly dose over the last 4 weeks of treatment was 182.20 +/- 118.11 IU/kg/wk, compared with 166.14 +/- 109.85 IU/kg/wk for epoetin alpha (95% CI: -3.21 to 35.34 IU/kg/wk). The most commonly reported AEs (> 5% of patients) were infections and infestations (12.5% and 12.8% of patients treated with epoetin zeta and epoetin alpha, respectively) and vascular disorders (8.5% and 8.9%, respectively). No patients developed neutralizing anti-erythropoietin antibodies. CONCLUSIONS: Epoetin zeta, administered intravenously, is therapeutically equivalent to epoetin alpha in the correction of low Hb concentration in patients with CKD undergoing haemodialysis. No unexpected AEs were seen and both epoetin zeta and epoetin alpha were well tolerated. PMID- 18394267 TI - Conference report on public health and clinical guidelines for anthrax. AB - On March 13-14, 2006, a meeting on anthrax, sponsored by the Centers for Disease Control and Prevention (CDC) in collaboration with the Southeastern Center for Emerging Biologic Threats, was held at Emory University in Atlanta, Georgia, USA. The meeting's agenda included discussion of postexposure prophylaxis (PEP), screening and evaluation, and treatment of the various manifestations of human anthrax. The goal was to convene subject matter experts for a review of research developments and clinical experience with anthrax prophylaxis and treatment and to make consensus recommendations for updating guidelines for PEP, treatment, and clinical evaluation of patients with anthrax. A 2001 conference on guidelines for anthrax has previously been summarized in this journal. This article summarizes the meeting's presentations and discussion. Consensus recommendations are summarized in the Table. Updated CDC guidelines for treatment and prophylaxis of anthrax will be published in detail in other CDC publications and are available on CDC's website at http://www.bt.cdc.gov/agent/anthrax/index.asp. PMID- 18394269 TI - Bluetongue epidemiology in the European Union. AB - Bluetongue (BT) is a reportable disease of considerable socioeconomic concern and of major importance in the international trade of animals and animal products. Before 1998, BT was considered an exotic disease in Europe. From 1998 through 2005, at least 6 BT virus strains belonging to 5 serotypes (BTV-1, BTV-2, BTV-4, BTV-9, and BTV-16) were continuously present in the Mediterranean Basin. Since August 2006, BTV-8 has caused a severe epizootic of BT in northern Europe. The widespread recrudescence and extension of BTV-8 infections in northern Europe during 2007 suggest that requirements for BTV establishment may now be fulfilled in this area. In addition, the radial extension of BTV-8 across Europe increases the risk for an encounter between this serotype and others, particularly those that occur in the Mediterranean Basin, where vector activity continues for more of the year. This condition increases the risk for reassortment of individual BTV gene segments. PMID- 18394270 TI - Potential use of antiviral agents in polio eradication. AB - In 1988, the World Health Assembly launched the Global Polio Eradication Initiative, which aimed to use large-scale vaccination with the oral vaccine to eradicate polio worldwide by the year 2000. Although important progress has been made, polio remains endemic in several countries. Also, the current control measures will likely be inadequate to deal with problems that may arise in the postpolio era. A panel convoked by the National Research Council concluded that the use of antiviral drugs may be essential in the polio eradication strategy. We here report on a comparative study of the antipoliovirus activity of a selection of molecules that have previously been reported to be inhibitors of picornavirus replication and discuss their potential use, alone or in combination, for the treatment or prophylaxis of poliovirus infection. PMID- 18394271 TI - Determination of oseltamivir quality by colorimetric and liquid chromatographic methods. AB - We developed a colorimetric and chromatographic assay for oseltamivir to assess the authenticity of Tamiflu (F. Hoffmann-La Roche Ltd., Basel, Switzerland) because of a growing concern about counterfeit oseltamivir. The colorimetric assay is quantitative and relies on an extractable colored ion-pair complex of oseltamivir with Congo red or bromochlorophenol blue. The reverse-phase chromatographic assay uses an alkaline mobile phase with UV detection. Both methods were evaluated for variability and selectivity and subsequently applied to batches of oseltamivir products acquired through the Internet. The Congo red test showed greater assay sensitivity, linearity, and accuracy. Colorimetric and chromatographic analysis showed all batches of oseltamivir product were within +/ 15% of the stated amount of active ingredient. PMID- 18394272 TI - Clonal population of flucytosine-resistant Candida tropicalis from blood cultures, Paris, France. AB - Candida tropicalis is a diploid ascomycetes yeast responsible for 4%-24% of candidemia. Resistance to flucytosine is rarely described for this species but was observed for 45 (35%) of 130 C. tropicalis isolates recovered from blood cultures in the Paris area in a 4-year survey. The aims of this study were to test the hypothesis that the flucytosine-resistant isolates could represent a subgroup and to determine the relationship between epidemiologic and genomic data. Epidemiologic data and gene sequences were analyzed, and molecular typing was performed. Our results suggest that a clone of flucytosine-resistant isolates, associated with malignancies and a lower mortality than that for other C. tropicalis isolates, is widespread in the Paris area. We propose the analysis of 2 polymorphic microsatellite markers coupled with URA3 sequencing to track the clone. PMID- 18394273 TI - Emericella quadrilineata as cause of invasive aspergillosis. AB - We noted a cluster of 4 cases of infection or colonization by Emericella spp., identified by sequence-based analysis as E. quadrilineata. Sequence-based analysis of an international collection of 33 Emericella isolates identified 12 as E. nidulans, all 12 of which had previously been identified by morphologic methods as E. nidulans. For 12 isolates classified as E. quadrilineata, only 6 had been previously identified accordingly. E. nidulans was less susceptible than E. quadrilineata to amphotericin B (median MICs 2.5 and 0.5 mg/L, respectively, p<0.05); E. quadrilineata was less susceptible than E. nidulans to caspofungin (median MICs, 1.83 and 0.32 mg/L, respectively, p<0.05). These data indicate that sequence-based identification is more accurate than morphologic examination for identifying Emericella spp. and that correct species demarcation and in vitro susceptibility testing may affect patient management. PMID- 18394274 TI - Control measures used during lymphogranuloma venereum outbreak, Europe. AB - To assess the response to the reemergence of lymphogranuloma venereum, we conducted a cross-sectional survey by administering a structured questionnaire to representatives from 26 European countries. Responses were received from 18 countries. The ability to respond quickly and the measures used for outbreak detection and control varied. Evidence-based criteria were not consistently used to develop recommendations. We did not develop criteria to determine the effectiveness of the recommendations. The degree of preparedness for an unexpected outbreak, as well as the ability of countries to respond quickly to alerts, varied, which indicates weaknesses in the ability to control an outbreak. More guidance is needed to implement and evaluate control measures used during international outbreaks. PMID- 18394275 TI - Beta-herpesviruses in febrile children with cancer. AB - We conducted a cross-sectional study of beta-herpesviruses in febrile pediatric oncology patients (n = 30), with a reference group of febrile pediatric solid organ transplant recipients (n = 9). One (3.3%) of 30 cancer patients and 3 (33%) of 9 organ recipients were PCR positive for cytomegalovirus. Four (13%) of 30 cancer patients and 3 (33%) of 9 transplant recipients had human herpesvirus 6B (HHV-6B) DNAemia, which was more common within 6 months of initiation of immune suppression (4 of 16 vs. 0 of 14 cancer patients; p = 0.050). HHV-6A and HHV-7 were not detected. No other cause was identified in children with HHV-6B or cytomegalovirus DNAemia. One HHV-6B-positive cancer patient had febrile disease with concomitant hepatitis. Other HHV-6B-positive children had mild "viral" illnesses, as did a child with primary cytomegalovirus infection. Cytomegalovirus and HHV-6B should be included in the differential diagnosis of febrile disease in children with cancer. PMID- 18394276 TI - Seroprevalence and risk factors for human herpesvirus 8 infection, rural Egypt. AB - To determine whether human herpesvirus 8 (HHV-8) is associated with schistosomal and hepatitis C virus infections in Egypt, we surveyed 965 rural household participants who had been tested for HHV-8 and schistosomal infection (seroprevalence 14.2% and 68.6%, respectively, among those <15 years of age, and 24.2% and 72.8%, respectively, among those > or =15 years of age). Among adults, HHV-8 seropositivity was associated with higher age, lower education, dental treatment, tattoos, > or =10 lifetime injections, and hepatitis C virus seropositivity. In adjusted analyses, HHV-8 seropositivity was associated with dental treatment among men (odds ratio [OR] 2.4, 95% confidence interval [CI] 1.1 5.2) and hepatitis C virus seropositivity among women (OR 3.3, 95% CI 1.4-7.9). HHV-8 association with antischistosomal antibodies was not significant for men (OR 2.1, 95% CI 0.3-16.4), but marginal for women (OR 1.5, 95% CI 1.0-2.5). Our findings suggest salivary and possible nosocomial HHV-8 transmission in rural Egypt. PMID- 18394277 TI - Retrospective analysis of monkeypox infection. AB - Serologic cross-reactivity between orthopoxviruses is a substantial barrier to laboratory diagnosis of specific orthopoxvirus infections and epidemiologic characterization of disease outbreaks. Historically, time-consuming and labor intensive strategies such as cross-adsorbed neutralization assays, immunofluorescence assays, and hemagglutination-inhibition assays have been used to identify orthopoxvirus infections. We used cross-adsorption to develop a simple and quantitative postadsorption ELISA for distinguishing between monkeypox and vaccinia infections. Despite the difficulty of diagnosing clinically inapparent monkeypox in previously vaccinated persons, this technique exhibited 100% sensitivity and 100% specificity for identifying clinically overt monkeypox infection irrespective of vaccination history. We also describe a Western blot technique in which up to 3 diagnostic bands may be used to distinguish between vaccinia and monkeypox infection. The techniques described provide independent diagnostic tests suitable for retrospective analysis of monkeypox outbreaks. PMID- 18394278 TI - Wild ducks as long-distance vectors of highly pathogenic avian influenza virus (H5N1). AB - Wild birds have been implicated in the expansion of highly pathogenic avian influenza virus (H5N1) outbreaks across Asia, the Middle East, Europe, and Africa (in addition to traditional transmission by infected poultry, contaminated equipment, and people). Such a role would require wild birds to excrete virus in the absence of debilitating disease. By experimentally infecting wild ducks, we found that tufted ducks, Eurasian pochards, and mallards excreted significantly more virus than common teals, Eurasian wigeons, and gadwalls; yet only tufted ducks and, to a lesser degree, pochards became ill or died. These findings suggest that some wild duck species, particularly mallards, can potentially be long-distance vectors of highly pathogenic avian influenza virus (H5N1) and that others, particularly tufted ducks, are more likely to act as sentinels. PMID- 18394279 TI - Rapid typing of transmissible spongiform encephalopathy strains with differential ELISA. AB - The bovine spongiform encephalopathy (BSE) agent has been transmitted to humans, leading to variant Creutzfeldt-Jakob disease. Sheep and goats can be experimentally infected by BSE and have been potentially exposed to natural BSE; however, whether BSE can be transmitted to small ruminants is not known. Based on the particular biochemical properties of the abnormal prion protein (PrPsc) associated with BSE, and particularly the increased degradation induced by proteinase K in the N terminal part of PrPsc, we have developed a rapid ELISA designed to distinguish BSE from other scrapie strains. This assay clearly discriminates experimental ovine BSE from other scrapie strains and was used to screen 260 transmissible spongiform encephalopathy (TSE)-infected small ruminant samples identified by the French active surveillance network (2002/2003). In this context, this test has helped to identify the first case of natural BSE in a goat and can be used to classify TSE isolates based on the proteinase K sensitivity of PrPsc. PMID- 18394280 TI - Hemorrhagic fever with renal syndrome caused by 2 lineages of Dobrava hantavirus, Russia. AB - Dobrava-Belgrade virus (DOBV) is a European hantavirus that causes hemorrhagic fever with renal syndrome (HFRS); case-fatality rates in Balkan countries are as high as 12%. To determine causative agents, we examined 126 cases of DOBV associated HFRS in central and southern European Russia. In central Russia (Lipetsk, Voronezh, Orel regions), outbreaks were caused by a DOBV variant (DOBV Aa) carried by Apodemus agrarius. In southern Russia (Sochi district), where HFRS is endemic, HFRS cases were caused by a new DOBV variant (DOBV-Ap), found in A. ponticus, a novel hantavirus natural host. Both viruses, DOBV-Aa/Lipetsk and DOBV Ap/Sochi, were isolated through Vero E6 cells, genetically characterized, and used for serotyping of the HFRS patients' serum. The clinical severity of HFRS caused by DOBV-Aa resembles that of HFRS caused by Puumala virus (mild to moderate); clinical severity of disease caused by DOBV-Ap infections is more often moderate to severe. PMID- 18394281 TI - Multiple sublineages of influenza A virus (H5N1), Vietnam, 2005-2007. AB - Phylogenetic analysis of influenza subtype H5N1 viruses isolated from Vietnam during 2005-2007 shows that multiple sublineages are present in Vietnam. Clade 2.3.4 viruses have replaced clade 1 viruses in northern Vietnam, and clade 1 viruses have been detected in southern Vietnam. Reassortment between these 2 sublineages has also occurred. PMID- 18394282 TI - Reassortant avian influenza virus (H5N1) in poultry, Nigeria, 2007. AB - Genetic characterization of a selection of influenza virus (H5N1) samples, circulating in 8 Nigerian states over a 39-day period in early 2007, indicates that a new reassortant strain is present in 7 of the 8 states. Our study reports an entirely different influenza virus (H5N1) reassortant becoming predominant and widespread in poultry. PMID- 18394283 TI - Neuroinvasion by Mycoplasma pneumoniae in acute disseminated encephalomyelitis. AB - We report the autopsy findings for a 45-year-old man with polyradiculoneuropathy and fatal acute disseminated encephalomyelitis after having Mycoplasma pneumoniae pneumonia. M. pneumoniae antigens were demonstrated by immunohistochemical analysis of brain tissue, indicating neuroinvasion as an additional pathogenetic mechanism in central neurologic complications of M. pneumoniae infection. PMID- 18394284 TI - Chagas disease, France. AB - Chagas disease (CD) is endemic to Latin America; its prevalence is highest in Bolivia. CD is sometimes seen in the United States and Canada among migrants from Latin America, whereas it is rare in Europe. We report 9 cases of imported CD in France from 2004 to 2006. PMID- 18394285 TI - Human thelaziasis, Europe. AB - Thelazia callipaeda eyeworm is a nematode transmitted by drosophilid flies to carnivores in Europe. It has also been reported in the Far East in humans. We report T. callipaeda infection in 4 human patients in Italy and France. PMID- 18394286 TI - Rabies virus in raccoons, Ohio, 2004. AB - In 2004, the raccoon rabies virus variant emerged in Ohio beyond an area where oral rabies vaccine had been distributed to prevent westward spread of this variant. Our genetic investigation indicates that this outbreak may have begun several years before 2004 and may have originated within the vaccination zone. PMID- 18394287 TI - Single nucleotide polymorphism typing of Bacillus anthracis from Sverdlovsk tissue. AB - A small number of conserved canonical single nucleotide polymorphisms (canSNP) that define major phylogenetic branches for Bacillus anthracis were used to place a Sverdlovsk patient's B. anthracis genotype into 1 of 12 subgroups. Reconstruction of the pagA gene also showed a unique SNP that defines a new lineage for B. anthracis. PMID- 18394288 TI - Human Mycobacterium bovis infection and bovine tuberculosis outbreak, Michigan, 1994-2007. AB - Mycobacterium bovis is endemic in Michigan's white-tailed deer and has been circulating since 1994. The strain circulating in deer has remained genotypically consistent and was recently detected in 2 humans. We summarize the investigation of these cases and confirm that recreational exposure to deer is a risk for infection in humans. PMID- 18394289 TI - Mycobacterium avium lymphadenopathy among children, Sweden. AB - We studied Mycobacterium avium lymphadenopathy in 183 Swedish children (<7 years of age) from 1998 through 2003. Seasonal variation in the frequency of the disease, with a peak in October and a low point in April, suggests cyclic environmental factors. We also found a higher incidence of the disease in children who live close to water. PMID- 18394290 TI - Kala-azar epidemiology and control, southern Sudan. AB - Southern Sudan is one of the areas in eastern Africa most affected by visceral leishmaniasis (kala-azar), but lack of security and funds has hampered control. Since 2005, the return of stability has opened up new opportunities to expand existing interventions and introduce new ones. PMID- 18394291 TI - Etymologia - Leishmaniasis. AB - Disease caused by protozoan parasites of the genus Leishmania, named in 1901 for British Army doctor William Leishman, who developed a stain to detect the agent. It is transmitted by the bite of certain species of sand fly, including the genus Lutzomyia in the New World and Phlebotomus in the Old World. PMID- 18394292 TI - Dengue virus type 4, Manaus, Brazil. AB - We report dengue virus type 4 (DENV-4) in Amazonas, Brazil. This virus was isolated from serum samples of 3 patients treated at a tropical medicine reference center in Manaus. All 3 cases were confirmed by serologic and molecular tests; 1 patient was co-infected with DENV-3 and DENV-4. PMID- 18394293 TI - Rat-to-elephant-to-human transmission of cowpox virus. PMID- 18394294 TI - Avian influenza knowledge among medical students, Iran. PMID- 18394295 TI - Lorraine strain of Legionella pneumophila serogroup 1, France. PMID- 18394296 TI - Bluetongue in captive yaks. PMID- 18394297 TI - Murine typhus, Algeria. PMID- 18394298 TI - Natural co-infection with 2 parvovirus variants in dog. PMID- 18394299 TI - WU polyomavirus infection in children, Germany. PMID- 18394300 TI - Hepatitis E, Central African Republic. PMID- 18394301 TI - Rickettsia sibirica subsp. mongolitimonae infection and retinal vasculitis. PMID- 18394302 TI - Rickettsia felis in fleas, France. PMID- 18394303 TI - Novel nonstructural protein 4 genetic group in rotavirus of porcine origin. PMID- 18394304 TI - PorB2/3 protein hybrid in Neisseria meningitidis. PMID- 18394305 TI - West Nile virus in birds, Argentina. PMID- 18394306 TI - Clostridium difficile surveillance trends, Saxony, Germany. PMID- 18394312 TI - CD 271 (P75 neurotrophin receptor). AB - P75NTR (or CD271) is a member of the Tumor Necrosis Factor receptor (TNFR) super family of transmembrane proteins that share significant homology in their extracellular domains. Subsets of TNF receptors, including CD271, have a cytoplasmic death domain, although CD271 has unique intracellular structure and downstream signaling partners. CD271 is also differentiated from other members of the TNFR receptor family in that it binds pro and mature neurotrophins and affects the growth, differentiation and death of the nervous system. The ligands for CD271 are neurotrophins, which are Nerve Growth Factor (NGF), Brain-Derived Growth factor (BDNF), Neurotrophin 3 (NT3) and Neurotrophin 4/5 (NT4/5). Recent studies have provided evidence that CD271 also serves as a receptor for the pro forms of these neurotrophins. PMID- 18394313 TI - From single gene to integrative molecular concept MAPS: pitfalls and potentials of microarray technology. AB - Microarray experiments have a large variety of applications and several important achievements have been obtained by means of this technology, especially within the field of whole genome expression profiling, which undoubtedly is the most diffused world-wide. Nevertheless, care must be taken in unconditionally applying such high-throughput techniques and in extracting/interpreting their results. Both the validity and the reproducibility of microarray-based clinical research have recently been challenged. Pitfalls and potentials of the microarray technology for gene expression profiling are critically reviewed in this paper. PMID- 18394314 TI - Toll-like receptors. AB - Toll-like receptors are a family of transmembrane receptors responsible for recognition and initiation of a response to invading microbes by the immune system. As part of the innate immune system, Toll-like receptors recognise pathogen-associated molecular patterns, highly conserved components that are essential to microbial function. Some of ten toll-like receptors identified in humans are able to recognise several pathogen-associated molecular patterns. PMID- 18394315 TI - Reproducibility of atopy patch tests with food and inhalant allergens. AB - Although atopy patch tests (APT) seem a valuable additional tool in the diagnostic work-up for food allergy in children with atopic eczema/dermatitis syndrome, the immunopathology and some technical aspects of testing remain controversial. Few published data are available on the reproducibility of APT with inhalants and only two studies include fresh food allergens. In this study we therefore investigated the reproducibility of duplicate APT (left versus right side of the back) with native and commercially available food (cow s milk, hen s egg, tomato, wheat flour) and with inhalant allergens (Dermatophagoides pteronyssinus and mixed grasses) in a large unselected population of children. We tested a population of 277 Italian school children with three APT allergens: fresh food (cow s milk, hen s egg, tomato and wheat flour), standardised food allergens in petrolatum (the same four foods) and standardised inhalant allergens routinely used for skin prick testing. For the four food allergens (applied in the natural form or as the standardised commercial preparation) from one- to three quarters of the APT gave positive results on one side and negative reactions on the opposite side (Cohen s K coefficient between 0.38, fresh tomato and 0.81, fresh cow s milk). Conversely, APT with inhalant allergens were invariably reproducible (Cohen s K = 1.00). The possible technical and immunologic reasons explaining why reproducibility of APT differed for the two types of allergens await an answer from extensive controlled studies. PMID- 18394316 TI - Vitamin K and D association stimulates in vitro osteoblast differentiation of fracture site derived human mesenchymal stem cells. AB - There is growing interest in osteoinductive agents for fracture healing especially in patients with non-union or delayed-union fractures. The aim of the present study is the assessment of the association of Vitamins D3 and K1 on proliferation and differentiation of human mesenchymal stem cells (hMSCs) derived from fracture sites in view of a possible clinical use. The synergic effect of Vitamin D3 and Vitamin K2 in preventing osteoporosis has been documented in clinical practice; however no reports investigating this association for fracture healing are present. Our data show a different outcome on cell proliferation linked to the different timing of drug administration as well as a synergic effect of the two vitamins on cell differentiation. The high level of osteocalcin and carboxylated osteocalcin detected in hMSCs treated with the association of the two vitamins in comparison with controls and with single vitamin administration underline the differentiation of these cells into osteoblastic phenotype. Our results indicate for the first time that vitamin D3 and K1 association is able to modulate in vitro the differentiation towards osteoblastic phenotype of hMSCs derived from fracture sites, thus offering clinicians a promising and low-cost strategy for reparative osteogenesis. PMID- 18394317 TI - A forest bathing trip increases human natural killer activity and expression of anti-cancer proteins in female subjects. AB - We previously reported that forest bathing trips enhanced human NK activity, number of NK cells, and intracellular anti-cancer proteins in lymphocytes, and that the increased NK activity lasted for more than 7 days after the trip in male subjects. In the present study, we investigated the effect of forest bathing trip on human NK activity in female subjects. Thirteen healthy nurses, age 25-43 years, professional career 4-18 years, were selected with informed consent. The subjects experienced a three-day/two-night trip to forest fields. On day 1, the subjects walked for two hours in the afternoon in a forest field; on day 2, they walked for two hours each in the morning and afternoon in two different forest fields; and on day 3, the subjects finished the trip and returned to Tokyo after drawing blood and completing a questionnaire. Blood and urine were sampled on the second and third days during the trip, and on days 7 and 30 after the trip. NK activity, numbers of NK and T cells, and granulysin, perforin, and granzymes A/B expressing lymphocytes in the blood samples, the concentrations of estradiol and progesterone in serum, and the concentrations of adrenaline and noradrenaline in urine were measured. Similar control measurements were made before the trip on a normal working day. The concentrations of phytoncides in the forests were measured. The forest bathing trip significantly increased NK activity and the numbers of NK, perforin, granulysin, and granzymes A/B-expressing cells and significantly decreased the percentage of T cells, and the concentrations of adrenaline and noradrenaline in urine. The increased NK activity lasted for more than 7 days after the trip. Phytoncides, such as alpha-pinene and beta-pinene were detected in forest air. These findings indicate that a forest bathing trip also increased NK activity, number of NK cells, and levels of intracellular anti cancer proteins in female subjects, and that this effect lasted at least 7 days after the trip. Phytoncides released from trees and decreased stress hormone levels may partially contribute to the increased NK activity. PMID- 18394318 TI - Anti-cyclic citrullinated peptide antibodies in patients affected by HCV-related arthritis. AB - Hepatitis C Virus (HCV) infection can induce immunological disorders with different clinical expressions such as arthritis, Sjogren Syndrome and various forms of vasculitis. Retrospectively, the prevalence of anti-Cyclic Citrullinated peptide antibodies (anti-CCP) in a group of patients affected by HCV-related arthritis with positivity for Rheumatoid Factor (RF) and the eventual correlations with RF and/or Anti-Nuclear Antibodies (ANA) and articular involvement were studied. Thirty patients with arthritis were selected from a population of 380 subjects affected by HCV infection. Each patient was evaluated by clinical examination (23 denoted poliarticular and 7 mono-oligoarticular involvement), by X-graphic aspects of joint involvement (8 patients presented joint erosions), by ANA, RF and anti-CCP positivity. Ten of the HCV-related arthritis patients (33.3 percent) presented positivity for anti-CCP, without significant correlation between such parameter and ANA, RF and articular involvement. Anti-CCP resulted positive in 4 out of the 8 patients with joint erosions, and only in 6 out of the 22 patients without joint erosions. Such frequencies analyzed by chi square resulted with no significant differences. Our patients presented an interesting prevalence of the positivity for anti-CCP. These data are cause to consider the specificity recently attributed to this parameter in the diagnosis of rheumatoid arthritis. PMID- 18394319 TI - Effect of a subgingival chlorhexidine chip on the clinical parameters and the levels of alkaline phosphatase activity in gingival crevicular fluid during the non-surgical treatment of periodontitis. AB - The main therapeutic approaches for inflammatory periodontal diseases include the mechanical treatment of root surfaces. Multi-center clinical trials have demonstrated that the adjunctive use of a chlorhexidine (CHX) chip is effective in improving clinical results compared to scaling and root planing (SRP) alone. However, some recent studies failed to confirm these clinical results, nor have any data been reported regarding the capability of the CHX chip in affecting the activity of alkaline phosphatase (ALP) in the gingival crevicular fluid (GCF). This enzyme has been related to a condition of destructive activity of periodontitis. The aim of this study is to provide further data on the clinical and biochemical effects of CHX chips when used as an adjunct to SRP. Eighty-two systemically healthy patients, aged 31-63, with moderate and advanced periodontitis were recruited from the departments of Periodontology of the University of Chieti. In each patient 2 experimental sites, located in two symmetric quadrants, were chosen with a probing depth of > or = 5 mm and bleeding on probing. The 2 sites were selected randomly at the split-mouth level; control sites received SRP alone, and test sites SRP plus 1 CHX chip. Clinical indices, including probing depth (PPD), clinical attachment level (CAL), bleeding on probing (BOP), and the ALP activity in GCF were evaluated at baseline and after 6 months. Alkaline phosphatase activity was assayed spectrophotometrically. The PPD and CAL were significantly lower at 6 months as compared to the baseline scores in both treatments (p less than 0.01). The PPD reduction was 2.7 mm in the CHX+SRP group and 1.9 mm in the SRP alone group. The CHX+SRP group showed a significantly greater gain of clinical attachment (mean: 1.4 mm) in comparison with the SRP group (mean: 0.9; p less than 0.05). No differences were observed in the decrease of the percent of BOP-positive sites between the experimental groups. Conversely, the CHX+SRP group underwent a significantly greater decrease (p less than 0.01) of the GCF-ALP activity 6 months after treatment in comparison with the SRP alone group. The adjunctive use of the CHX chip resulted in a significant improvement of pocket reduction and clinical attachment gain as compared with SRP alone. These results were concomitant with a significantly greater reduction of the GCF-ALP activity levels. PMID- 18394320 TI - Quality of life in non-allergic rhinitis depends on the predominant inflammatory cell type. AB - Three main types of inflammatory Non-Allergic Rhinitis (NAR) have been defined: NAR infiltrated by eosinophils (NARES), by mast cells (NARMA), and by neutrophils (NARNE). In the absence of studies that investigated the Quality of Life (QoL) in NAR, the present work is aimed at evaluating the Quality of Life of patients with NARES, NARMA, and NARNE. One hundred thirty one (131) NAR patients were prospectively and consecutively evaluated: 54 patients with NARES, 38 with NARMA, and 39 with NARNE. Their history, nasal infiltration and rhinomanometry were characterized, and Quality of Life (using 2 instruments) was evaluated, and associated to clinical and histological features. Quality of Life was significantly different in the 3 groups (p less than 0.001); NARES patients had the worst Quality of Life. Nasal resistances were significantly higher in the NARES group. Significant associations were shown in NARES patients between Quality of Life and nasal function. This study provides the first evidence that Quality of Life is impaired in NAR as well as in allergic rhinitis. Furthermore, Quality of Life impairment differs among the various forms of NAR, and there is a correlation with the cellular infiltrating type. PMID- 18394321 TI - Subjective and psychopathological response in patients under different antipsychotic treatments: are there differences in real clinical practice? AB - The aim of this study is to investigate whether subjective well-being in patients under treatment with typical (ATPs) and atypical antipsychotic (ATPsA) compounds can be compared with the improvement of psychopathological state and to verify if both variables correlate to adherence to treatment. We assessed 106 consecutive patients receiving ATPs or ATPsA in the University Psychiatric Ward of L?Aquila, according to DSM-IV diagnosis of Schizophrenia and Bipolar Disorder. Psychopathological state was assessed by Brief Psychiatric Rating Scale-4.0 version (BPRS), adherence to treatment and subjective well-being was assessed by Drug Attitude Inventory (DAI-10) and Subjective Well-being under Neuroleptics (SWN), respectively. BPRS and DAI-10 were administered on admission (T0) and at the end of recovery (T1). The subjects enrolled in this study were divided into 2 groups according to ATP prescribed. We observed an improvement of BPRS and SWN total scores in each group, and increasing scores in DAI-10, from admission to discharge, both in total samples and in each group. There were statistical differences between the patients receiving ATPs and those receving ATPsA regardindg the SWN total score and its different dimensions. This study emphasizes that patients receiving ATPsA show better subjective response compared with patients undergoing ATP treatment, although the adherence to pharmacotherapy and clinical improvement do not differ between the groups. PMID- 18394322 TI - Thermal strain in fire fighters while wearing task-fitted versus en 469:2005 protective clothing during a prolonged rescue drill. AB - Fire fighters are normally overprotected during their working hours because of the tendency to keep the personal protection level sufficiently high in case of the worst possible scenarios. This study investigated the effects of task-fitted protective clothing on thermal strain in fire fighters as compared to EN 469:2005 protective clothing during a prolonged (2 1/2 hrs) job-related rescue drill under neutral and hot climates. The subjects were 23 healthy, physically fit professional male fire fighters aged 26-44 years. Measurements included cardiovascular and thermal responses and subjective assessments. Wearing task fitted clothing during rescue tasks in a neutral climate considerably reduced total thermal and cardiovascular strain in prolonged rescue work. The fire fighters also perceived physical work as significantly harder on average, and reported more intense subjective discomfort while wearing EN 469:2005 as compared to task-fitted clothing. PMID- 18394323 TI - Development of a test method against hot alkaline chemical splashes. AB - High temperature alkaline chemical liquids have caused injuries and hazardous situations in Finnish pulp manufacturing mills. There are no requirements and/or test method standards concerning protection against high temperature alkaline chemical splashes. This paper describes the test method development process to test and identify materials appropriate for hot liquid chemical hazard protection. In the first phase, the liquid was spilled through a stainless steel funnel and the protection performance was evaluated using a polyvinyl chloride (PVC) film under the test material. After several tentative improvements, a graphite crucible was used for heating and spilling the chemical, and a copper coated K-type thermometer with 4 independent measuring areas was designed to measure the temperature under the material samples. The thermometer was designed to respond quickly so that peak temperatures could be measured. The main problem was to keep the spilled amount of chemical constant, which unfortunately resulted in significant variability in data. PMID- 18394324 TI - Protection from steam at high pressures: development of a test device and protocol. AB - Extensive use of pressurized steam in the oil and gas sectors has led to incidents where workers were seriously injured. In this study a test device and procedure to measure heat transfer through fabrics during steam exposure were developed and evaluated. Several factors were considered while designing the test device to simulate work site conditions. Fabrics were exposed to steam at 2 distances (50 and 100 mm) and 2 pressures (207 and 69 kPa). Theoretical considerations included heat and mass transfer, and fabric structure and performance properties. The test device and procedure differentiated well among both fabrics and exposure conditions. For all fabrics, maximum heat transfer was observed at highest steam pressure and shortest distance. Laminated and coated fabrics performed better than a fabric without such treatments. PMID- 18394325 TI - Optimizing the performance of phase-change materials in personal protective clothing systems. AB - Phase-change materials (PCM) can be used to reduce thermal stress and improve thermal comfort for workers wearing protective clothing. The aim of this study was to investigate the effect of PCM in protective clothing used in simulated work situations. We hypothesized that it would be possible to optimize cooling performance with a design that focuses on careful positioning of PCM, minimizing total insulation and facilitating moisture transport. Thermal stress and thermal comfort were estimated through measurement of body heat production, body temperatures, sweat production, relative humidity in clothing and subjective ratings of thermal comfort, thermal sensitivity and perception of wetness. Experiments were carried out using 2 types of PCM, the crystalline dehydrate of sodium sulphate and microcapsules in fabrics. The results of 1 field and 2 laboratory experimental series were conclusive in that reduced thermal stress and improved thermal comfort were related to the amount and distribution of PCM, reduced sweat production and adequate transport of moisture. PMID- 18394326 TI - Performance of firefighters' protective clothing after heat exposure. AB - Heat and mechanical protection properties of 6 fabric combinations commonly used in firefighters' protective clothing were assessed before and after different heat treatment. It was shown that after heat exposure, the values obtained were generally lower than in the original state. The mechanical properties of the materials were more affected by heat than by heat protective properties. In 2 cases, degradation started before a visible change in the material could be observed, which might be potentially dangerous for the end user who will not realize the alteration of the material. PMID- 18394327 TI - Evaluation of the flexibility of protective gloves. AB - Two mechanical methods have been developed for the characterization of the flexibility of protective gloves, a key factor affecting their degree of usefulness for workers. The principle of the first method is similar to the ASTM D 4032 standard relative to fabric stiffness and simulates the deformations encountered by gloves that are not tight fitted to the hand. The second method characterizes the flexibility of gloves that are worn tight fitted. Its validity was theoretically verified for elastomer materials. Both methods should prove themselves as valuable tools for protective glove manufacturers, allowing for the characterization of their existing products in terms of flexibility and the development of new ones better fitting workers' needs. PMID- 18394328 TI - Dry and wet heat transfer through clothing dependent on the clothing properties under cold conditions. AB - The purpose of this study was to investigate the effect of moisture on the heat transfer through clothing in relation to the water vapour resistance, type of underwear, location of the moisture and climate. This forms part of the work performed for work package 2 of the European Union THERMPROTECT project. Thermal manikin results of dry and wet heat loss are presented from different laboratories for a range of 2-layer clothing with similar dry insulations but different water vapour permeabilities and absorptive properties. The results obtained from the different manikins are generally consistent with each other. For each climate, total wet heat loss is predominately dependent on the permeability of the outer layer. At 10 degrees C, the apparent evaporative heat loss is markedly higher than expected from evaporation alone (measured at 34 degrees C), which is attributed to condensation within the clothing and to increased conductivity of the wet clothing layers. PMID- 18394329 TI - Variable-thermoinsulation garments with a microprocessor temperature controller. AB - This paper presents the concept of active variable thermoinsulation clothing for users working in low temperatures. Those garments contain heating inserts regulated by a microprocessor temperature controller. This paper also presents the results of tests carried out on the newly designed garments. PMID- 18394331 TI - Protection provided by clothing and textiles against potential hazards in the operating theatre. AB - The typical hospital and operating theatre present multiple potential hazards to both workers and patients, and protection against some of these is provided through use of various forms of clothing and textiles. While many standards exist for determining the performance of fabrics, most tests are conducted under laboratory conditions and against a single hazard. This paper provides an overview of selected developments in the principal properties of fabrics and garments for use in these workplaces, identifies the key standards, and suggests topics for further investigation. PMID- 18394330 TI - Numerical simulations of heat and moisture transport in thermal protective clothing under flash fire conditions. AB - A numerical model of heat and moisture transport in thermal protective clothing during exposure to a flash fire was introduced. The model was developed with the assumption that textiles are treated as porous media. The numerical model predictions were compared with experimental data from different fabric systems and configurations. Additionally, with the introduction of a skin model, the parameters that affect the performance of thermal protective clothing were investigated. PMID- 18394332 TI - [Identification of a novel antigenic epitope on GST fusion-expressed and ORF2 encoded proteins of hepatitis E virus]. AB - AIM: To investigate the effect of a GST tag on the antigenic structure of GST fusion-expressed and ORF2-encoded recombinant proteins of hepatitis E virus (HEV). METHODS: The monoclonal antibodies (mAb) were prepared with a GST fusion protein, p166Chn-GST, which was derived from a Chinese HEV strain. Then they were tested by indirect ELISA, competition ELISA and Western blot with different GST fusion, His fusion or non-fusion recombinant proteins derived from HEV reference strains of all 4 genotypes and other non-HEV recombinant proteins. RESULTS: Three mAb named 1A8, 9B4 and 8H10 were obtained. All of them reacted to p166Chn-GST but did not react to GST. mAb 1A8 and 9B4 reacted to 4 p166-GST proteins of different HEV genotypes and 2 N- or C-terminal truncated p166Chn-GST proteins named p146Chn GST and p137Chn-GST, but they did not react to 4 p166-His proteins of different HEV genotypes and a non-fusion p179Chn protein. No detectable signals were found when 1A8 and 9B4 were subjected to HEV antigen competition ELISA or Western blot after SDS-PAGE. No cross reaction was observed between the two mAb and HEV irrelevant GST fusion proteins, either. CONCLUSION: A novel antigenic epitope recognized by mAb 1A8 and 9B4 appears on the GST fusion-expressed and ORF2 encoded HEV recombinant proteins and it is dependent on the conformational folding of both GST and HEV sequences. PMID- 18394333 TI - [Effect of high mobility group box 1 protein on proliferation and apoptosis and balance between Th1/Th2 and Tc1/Tc2 of lymphocytes in vitro]. AB - AIM: To explore whether extracellular high mobility group B1 protein (HMGB1) plays a role in regulation of lymphocyte-mediated immune. METHODS: Lymphocytes originated from mice spleens were stimulated with concentration gradient HMGB1 or combined with ConA in vitro. Then, the proliferation of lymphocytes was assayed with MTT, quantitative and qualitative analysis of apoptosis in lymphocytes and expression of CD3 and CD8 on cells surface and IL-4 and IFN-gamma within cells were assessed using flow cytometry. Levels of interleukin-2 (IL-2) and soluble IL 2R (sIL-2R) of culture supernatant were assayed by ELISA. RESULTS: (1) HMGB1 regulated the proliferation of lymphocytes in a time- and dose-dependent manner, but did not influence the apoptosis of lymphocytes. (2) The ratio of Th to Tc increased from 2.6:1 at culture-hour 0 to 5-7:1 at culture-hour 12 and 24, but that did not change with the stimulation of HMGB1. There were no significant difference in the population and percentage of Th1 and Th2, as well as Tc1 and Tc2, when stimulated with HMGB1. However, HMGB1 in concentrations of 10 and 100 microg/L favored Th1 differentiation, and 1 and 10 microg/L HMGB1 favored Tc1 differentiation. (3) Levels of IL-2 were increased, and sIL-2R decreased, significantly, when stimulated with 10 microg/L HMGB1 for 12 h, thus the ratio of IL-2 to sIL-2R was markedly higher, as compared with 0 and 1000 microg/L HMGB1 (P<0.05 or P<0.01). CONCLUSION: Low dose HMGB1 could be in favor of the bias of Th1 and Tc1, and improve the lymphocyte-mediated immune. PMID- 18394334 TI - [Effect and mechanism of curcumin on learning and memory dysfunction induced by gp120 in rats]. AB - AIM: To explore the effect and mechanisms of curcumin on learning and memory dysfunction induced by HIV-1 enveloped protein gp120. METHODS: The SD rats were treated with gp120 by intracerebroventricular (ICV) infusion imitating the HIV-1 associated dementia (HAD) animal model. Subsequently, we applied the water maze test to evaluate the effect of gp120 on the learning and memory dysfunction in rats. The SD rats were divided into six groups: control group, sham group, model group, low dose curcumin group, middle dose curcumin group and high dose curcumin group. Except control and sham group, the others four groups received slowly 5 microL/d gp120 which dissolved in artificial cerebrospinal fluid (ACSF) for 3 days. Since the fourth day, the rats of low, middle, high dose curcumin groups were treated with 50 mg/(kg.d), 100 mg/(kg.d), 200 mg/(kg.d) curcumin, respectively. The others groups were treated with redistilled water. The treatment lasted for 14 days. Subsequently, the water maze test and NMDA2BR immunohistochemical staining were applied to evaluate the effect of curcumin on the rats. RESULTS: The rats were treated with gp120 50 ng/d by ICV infusion for 3 days can imitate the HAD animal model. The Morris water maze (MWM) test showed that the rats in model group had longer escape latencies compared with those in control group (P<0.05) and that rats in low, middle, high dose curcumin groups had shorter escape latencies compared with those in model group (P<0.05), and low dose curcumin group was better than the other two groups (P<0.05). Immunohistochemical staining showed that the expressions of NMDA2BR in model group decreased compared with the control groups (P<0.01), while the expressions of NMDA2BR in low, middle and high dose curcumin groups increased compared with the model groups. CONCLUSION: The SD rats were treated with gp120 by ICV infusion imitating the HAD animal model. The curcumin can improve the learning and memory dysfunction induced by gp120, the mechanism may be related to against the downregulation the expression of NMDA2BR. PMID- 18394335 TI - [Construction of eukaryotic expression plasmid containing rat interleukin-10 gene and its expression in BRL cells in vitro]. AB - AIM: To construct eukaryotic expression vector of rat IL-10 gene and observe its expression in hepatocyte cell line BRL. METHODS: Total RNA was extracted from rat peripheral blood mononuclear cells. The full length coding region of IL-10 was amplified by RT- nested PCR and cloned into eukaryotic expression vector pcDNA3.0. The recombinant plasmid was transfected into BRL cells with either liposome Transfast or asialoglycoprotein receptor mediated liposome PEIjet-gal respectively. The expression of IL-10 mRNA was detected with PCR and that of IL 10 secreted from BRL cells transfected by liposome PEIjet-gal was detected with ELISA. RESULTS: The recombinant plasmid was identified and confirmed with digestion of restriction endonuclease and DNA sequencing. Receptor mediated liposome PEIjet-gal exhibited significantly higher transfection efficiency than liposome Transfast and higher level secretory IL-10 expressed in BRL cells. CONCLUSION: The eukaryotic expression vector of IL-10 gene was successfully constructed. Asialoglycoprotein receptor-mediated liposome had high transfection efficiency on hepatocytes, suggesting that it could be a potential hepatocyte targeting delivery system for IL-10 gene therapy. PMID- 18394336 TI - [Immunoregulation of IL-18 in mice with radiated damage]. AB - AIM: To explore the immunoregulation of IL-18 in the mice radiated by 60Co. METHODS: 32 C57 mice were radiated by 60Co and then treated by IL-18.2 weeks later, the transformation of lymphocytes, the ability of NK cells to kill tumor cells, the subtype of T cells and the content of IgG in serum were tested. RESULTS: IL-18 increased the function of lymphocyte transformation in 60Co radiated mice, enhanced the cytotoxic activity of NK cells against tumor cells of A375, U973 and KG1, and up-regulated the amount of CD4+T cells. However, the level of IgG in the serum of the radiated mice was not regulated by IL-18. CONCLUSION: IL-18 can enhance the immune function of the mice radiated by 60Co. PMID- 18394337 TI - [Effect of Apigenin on proliferation, cell cycle and apoptosis of mouse T cells in vitro]. AB - AIM: To study the effect of Apigenin (AP) on the proliferation, cell cycle and apoptosis of mouse T cells in vitro. METHODS: The lymphocytes were prepared from lymph nodes and thymus of mice. The effect of AP on the proliferation of T in response to ConA at different concentrations (25, 50, 100, 150, 200 micromol/L) was detected by MTT. Cell cycle was measured by PI staining and FCM. The effect of Apigenin and Apigenin with DEX on T cell apoptosis was measured by Annexin V FITC/PI double staining and FCM. The effect of different concentrations of AP cytotoxicity to T cells was measured by MTT. RESULTS: 25-200 micromol/L of AP didn't have cytotoxicity to T cells, but it had some inhibitory effect on T cells in response to ConA(P<0.01), arresting cell cycle at G0/G1 in a dose-dependent manner. Different concentrations of AP inhibited the apoptosis of T cells, especially those induced by DEX(P<0.01). CONCLUSION: AP can inhibit the proliferation of mouse T cells in response to ConA, arrest cell cycle at G0/G1 and inhibit the apoptosis of T cells. PMID- 18394338 TI - [Expression of NKG2D ligands on dendritic cells at different development stages and its effect on cytotoxicity of NK cells]. AB - AIM: To investigate the expression of NKG2D ligands on dendritic cells(DC) at different development stages and its effect on cytotoxicity of natural killer(NK) cells. METHODS: The monocytes were cultured into immature dendritic cells(iDC) and mature dendritic cells(mDC) with cytokines. NK cells were obtained from normal peripheral blood by CD56 antibody magnetic isolation.The expression of NKG2D ligands (MICA/B, ULBP1-3) was detected by flow cytometry. Cytotoxicity of NK cells and the NK cells blocked with anti-NKG2D mAbs against iDC and mDC was tested using LDH-releasing method. RESULTS: IDC and mDC were of typical morphology and phenotypes. MICA, MICB, ULBP1, and ULBP3 were expressed on iDC and their expression rate was (32.39+/-8.30)%, (17.75+/-3.40)%, (26.71+/-6.48)%, (38.37+/-6.89)%, respectively. MICA and ULBP3 were expressed on mDC and their expression rate was (7.81+/-3.33)% and (8.36+/-2.42)%, respectively, which was lower than that on mDC (P<0.01). At the each E:T ratio cytotoxicity of NK cells against iDC was stronger than that against mDC (P<0.01). cytotoxicity of NK cells blocked with anti-NKG2D mAb against iDC was decreased compared with that of NK cells unblocked (P<0.05) while cytotoxicity of NK cells blocked with anti-NKG2D mAb against mDC showed no decrease compared with that of NK cells unblocked (P>0.05). CONCLUSION: The expression of NKG2D ligands on iDC is higher than that on mDC, which plays an important role in the cytotoxic effect of NK cells against iDC, but has no effect on that against mDC. NKG2D-NKG2D ligands shows one of the molecular mechanisms that NK cells kill iDC selectively. PMID- 18394339 TI - [Artificial chaperone-assisted refolding of recombined chicken Interleukin-18 gene in E.coli]. AB - AIM: To study the technique of boosting the renaturation yield of rChIL-18 by using artificial molecular chaperone composed of cetyl trimethyl ammonium bromide (CTAB) and beta-cyclodextrin(beta-CD). METHODS: The recombinant plasmid of mChIL 18 prokaryotic expression was transformed into E.coli BL21 (DE3) strain and then induced by IPTG at 37 degrees Celsius. The recombinant mChIL-18 was expressed efficiently in inclusion bodies in E.coli. After crushed and washed, the inclusion bodies were thoroughly denatured with 6 mol/L of guanidine hydrochloride, and then the artificial molecular chaperone was used to promote protein refolding. After the rehabilitation of products was purified by bag filter, its activity was detected by lymphocyte proliferation assays. RESULTS: The SDS-PAGE analysis indicated the expressed ChIL-18 protein had molecular weight of 44000. The expressed product existed in the form of inclusion body.Two protein bands of Mr 44000 and 26000 appeared on SDS-PAGE gel. The percentage of renaturation was 42.54 with artificial molecular chaperone.The results of MTT assay showed the expression of ChIL-18 protein in E.coli BL21 (DE3) greatly induced the proliferation of chicken T lymphocytes. CONCLUSION: The artificial chaperone technique can obviously boost the renaturation yield of rChIL-18. The purified and expressed product of fusion chicken Interleukin-18 gene in E.coli have relativity high bioactivity. PMID- 18394340 TI - [Prokaryotic expression of the extracellular region of human CD1d and preparation of its polyclonal antibody]. AB - AIM: To explore the prokaryotic expression of the extracellular region of human CD1d (hCD1d) and prepare its polyclonal antibody. METHODS: The gene encoding the extracellular region of hCD1d was amplified by PCR and cloned into prokaryotic expression vector pET28, then expressed in E.coli BL21 (DE3) with IPTG induction. The recombinant protein was purified by Ni2+-NTA agarose column and then used as immunogen to immunize the mouse. The generated polyclonal antibody was evaluated by ELISA, Western blot and immunohistochemical staining (IHC), respectively. RESULTS: The recombinant extracellular region of hCD1d was successfully expressed and purified. The polyclonal antibody with high titer and high specificity was obtained, which could recognize the native hCD1d in the human small intestinal tissues. CONCLUSION: The recombinant extracellular region of hCD1d has been obtained. The antibody with high titer and high specificity against the extracellular region of hCD1d from the mouse has been successfully prepared, which lays a foundation for further research into the detection and functional study of CD1d. PMID- 18394341 TI - [Mass spectrometric analysis of peripheral blood mononuclear cells in HCV infected patients]. AB - AIM: To investigate the diversity of proteins' pattern of the peripheral blood mononuclear cells (PBMCs) in patients with hepatitis C virus (HCV) infection, identify the differentially expressed proteins and analyze their roles in the mechanism of chronic hepatitis C. METHODS: The total proteins from PBMCs in HCV infected patients (28) and healthy persons (10) were separated by the immobilized pH gradient-based 2-DE. The differentially expressed proteins were screened by PDQuest analysis software, and then identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: 2-DE results showed that the mean number of protein spots in HCV infected patients and healthy persons was 625 and 614. Twelve differential proteins were tested by MALDI-TOF MS, and 10 of them were identified. Some of the proteins participated in protein synthesis and degradation, signal transduction, metabolism, or cytoskeleton construction while some of them were proteins of virus. CONCLUSION: 2-DE pattern of PBMCs from HCV infected patients or healthy persons has been established and ten differentially expressed proteins has been characterized. Our study is helpful for clinical detection of HCV infection and further research into the mechanisms of chronic hepatitis C. PMID- 18394342 TI - [Immunocontraceptive effect of DNA vaccines of Lagurus lagurus zona pellucida 3 fusioned with C-terminal of HSP70]. AB - AIM: To investigate the immunocontraceptive effect of Lagurus lagurus zona pellucida 3 gene (LZP3) fused by Mycobacterium tuberculosis HSP70 and C-terminal of HSP70. METHODS: Two immunocontractive recombinant plasmids pcD-L-HSP70 and pcD L-HSP70C were constructed using LZP3. Meanwhile, pcD-L and pcD-ACLC which were effective immunocontraceptive vaccines were studied as control. RESULTS: After the LZP3 genes were expressed in the livers of mice with the introduction of hydronamic transfection instead of traditional HeLa cell transfection by RT-PCR, NIH mice were immunized with ZP3 DNA vaccines. The results of ELISA showed that all of the recombinants in mice elicited specific anti-ZP3 antibodies which were combined with the ZP3 of oocytes from the immunized mice in immunofluorescence assay. MTS assay showed that the lymphocytes in all groups were proliferated by stimulating the recombinant LZP3 with no disruption of follicular in ovaries, especially those in groups of pcD-ACLC and pcD-L-HSP70C (P<0.01). Antifertility experiments showed that three groups (besides pcD-L-HSP70) enhanced the sterile effects (P<0.05), especially those in the groups of pcD-ACLC and pcD-L-HSP70C (P<0.01). CONCLUSION: LZP3 fused by the C-terminal of mycobacterium tuberculosis heat shock protein HSP70 had a good immunocontraceptive effect while LZP3 fused by Mycobacterium tuberculosis heat shock protein HSP70 had a poor effect on birth control. So the C-terminal of Mycobacterium tuberculosis heat shock protein HSP70 might be a better candidate to deliver the adjuvant function in LZP3 DNA immunocontraceptive vaccination than the complete HSP70 molecule. PMID- 18394343 TI - [Gene cloning of ligand binding domain of porcine beta3 as FMDV receptor and preparation of its polyclonal antibody]. AB - AIM: To clone and express the ligand binding domain (LBD) cDNA of porcine integrin beta3 as foot-and-mouth disease virus (FMDV) receptor and prepare its polyclonal antibody. METHODS: The LBD cDNA of porcine beta3 was obtained from the lung tissue of pig infected with FMDV by RT-PCR, and the recombinant plasmid pGEM/beta3LBD was constructed. After digested with BamH I/Xho I, the beta3LBD fragment was subcloned into prokaryotic expression vector pGEX 4T-1. The recombinant expression plasmid pGEX/beta3LBD was constructed and transformed into E.coli BL21(DE3). The recombinant porcine beta3LBD protein was expressed after IPTG induction and purified from total protein of BL21(DE3). The rabbits from New Zealand were immunized with the purified fusion protein to prepare polyclonal antibody, which was identified by Western blot and ELISA. RESULTS: The 507 bp cDNA of porcine beta3LBD encoded a polypeptide of 169 amino acids. The similarity of nucleotide sequence beta3LBD between pigs and cattle, human being, chimpanzees, rhesus monkeys, horses, dogs, Norway rats, mice, chickens was 90.3%, 92.3%, 92.1%, 91.3%, 90.5%, 90.3%, 87.8%, 85.2%, 79.5%, respectively. The beta3LBD gene of mammals exhibited high sequence homology. The recombinant beta3LBD protein was expressed efficiently as inclusion body after IPTG induction and was approximately 44000. The titer of the polyclonal antibody against the purified beta3LBD protein was about 1:12 800 by ELISA. CONCLUSION: The gene cloning and expression of beta3LBD and the preparation of its polyclonal antibody lay a foundation for further research into the interaction of FMDV with beta3 subunit of porcine integrin. PMID- 18394344 TI - [Establishment of the magnetic separation enzyme immunofluorescence method for detecting insulin]. AB - AIM: To establish a convenient and sensitive magnetic separation enzyme immunofluorescence (MEIF) method for detecting human insulin. METHODS: Two monoclonal antibodies (mAbs) were conjugated with FITC and alkaline phosphatase (AP) respectively, which were incorporated magnetic solid phase separation. Magnetic beads were coupled with sheep anti-FITC antibody as solid phase and 4 Methylumbelliferyl-phosphoric acid (4-MUP) was used as substrate to set up MElF for detecting insulin. RESULTS: The sensitivity of this assay was 2.0 microIu/mL, the linear range was from 0 microIu/mL to 188.52 microIu/mL, and the intra-assay variation and inter-assay variation were 4.3%-5.2% and 2.6%-9.5%, respectively. The recovery rate of dilution was 92.6%-117% and the recovery rate of accession was 106%-121%.The result of the assay correlated well with that of magnetic enzyme chemiluminescence immunoassay system. CONCLUSION: The MEIF for detecting insulin is low at cost, sensitive, specific and stable, which can be widely used in clinical immune detection. PMID- 18394345 TI - [Expression of Foxp3 in CD4+CD25high Treg cells of neonatal cord blood]. AB - AIM: To investigate the amount of CD4+CD25high Treg cells and the expression of Foxp3 in neonatal cord blood, and to analyze the expression of Treg cells in neonates. METHODS: The mononuclear cells from cord blood or peripheral blood were isolated from neonatal cord blood (n=15) or healthy adult's peripheral blood (n=12) by density gradient centrifugation. Then they were stained with fluorescence labeled monoclonal antibodies for cell surface and intracellular protein. The amount of CD4+CD25high Treg cells and intercellular transcription factor Foxp3 were measured by flow cytometry. RESULTS: The amount of CD4+CD25high Treg cells in CD3+CD4+ T cells in cord blood was significantly higher than that in adult's peripheral blood (3.86%+/-1.63% vs 0.87%+/-0.74%, P<0.01), whereas the expression of Foxp3 in CD4+CD25high Treg cells in cord blood was markedly lower than in adult's blood (23.21%+/-8.9% vs 71.3%+/-11.6%, P<0.01). CONCLUSION: The low expression of Foxp3 in CD4+CD25high Treg cells in cord blood suggests Treg cells in neonates are not mature in suppression. PMID- 18394346 TI - [Expressions of interleukin 4 mRNA and interferonjgamma mRNA of condyloma acuminata]. AB - AIM: To study the expressions of interleukin 4 (IL-4) mRNA and interferon-gamma (IFN-gamma) mRNA of condyloma acuminata in order to obtain a clue as to the immunological mechanisms possibly relevant for warts regression or persistence. METHODS: In situ RT-PCR was employed in surgically removed warts of 22 recurrent patients, 14 non-recurrent patients and 15 normal controls, respectively. RESULTS: The expression of IFN-gamma mRNA in the non-recurrent warts was higher than in recurrent warts (P<0.01); but the expression of IL-4 mRNA in the recurrent warts was higher than in non-recurrent warts (P<0.01). CONCLUSION: A higher IFN-gamma mRNA expression of condyloma acuminata lesions might contribute to warts regression. PMID- 18394347 TI - [Clinical value of combined determination of serum and pleural effusion level of CEA,CYFRA21-1, TPS in the diagnosis of lung cancer]. AB - AIM: To study the clinical value of combined determination of carcinoembryonic antigen (CEA), cytokeratin-19-fragments (CYFRA21-1) and tissue polypeptide specific antigen (TPS) levels in both serum and pleural effusion in the diagnosis of lung cancer. METHODS: The levels of CEA, CYFRA21-1 and TPS in serum and pleural effusion were measured in 78 patients with lung cancer and 45 patients with benign lung disease by using electrochemiluminescence and ELISA methods respectively. RESULTS: The levels of CEA, CYFRA21-1 and TPS in pleural effusion of patients with lung cancer were much higher than those with benign lung disease (P<0.01). The levels of CEA and TPS in serum of patients with lung cancer were much higher than those with benign lung disease (P<0.05, P<0.01). It was also found that the levels of these tumor markers in pleural effusion were higher and more sensitive than those in sera, especially TPS. TPS showed the highest sensitivity in single tumor marker detection, and TPS+CYFRA21-1+ CEA showed the highest sensitivity and accuracy in combined tumor marker detection for diagnosis of lung cancer in pleural effusion. CONCLUSION: The combined detection of CEA, CYFRA21-1 and TPS in pleural effusion showed higher sensitivity, better accuracy and higher clinical value than those in serum for diagnosis of lung cancer. PMID- 18394348 TI - [The up-regulated expression of LAIR-1 in tumor patients PBMC]. AB - AIM: To investigate the expression of LAIR-1 in patients with tumors and the function of LAIR-1 in anticancer immunity. METHODS: A sandwich ELISA was employed to detect the serum sLAIR-1 from tumor patients and healthy individuals. The expression of LAIR-1 in CD4+ T cells, CD8+ T cells, NK cells and B cells isolated from the peripheral blood of cancer patients was examined by fluorescence staining and flow cytometry. RESULTS: The level of serum sLAIR-1 in tumor patients was higher than that in healthy individuals [4.6+/-3.2 microg/L vs 3.9+/ 3.0 microg/L, P<0.05]. NK cells, CD4+ T cells and CD8+ T cells expressed the up regulated LAIR-1. The ratio of CD4/CD8 in lung cancer patients decreased significantly while the percentage of B cells in cancer patients increased greatly compared with healthy individuals. CONCLUSION: The expression of LAIR-1 is up-regulated in tumor patients, which may contribute to cancer immune escape. PMID- 18394349 TI - [Inhibitory effect of huangqi and dangshen extraction with pacilitaxel on metastasis and angiogenesis on mouse Lewis lung carcinoma model]. AB - AIM: To study enhance effect of huangqi and dangshen extraction (Shenqi) on pacilitaxel inhibitory metastasis and angiogenesis on mouse Lewis lung carcinoma model. METHODS: Lewis lung carcinoma cells were inoculated into right hind footpad of C57BL/6 mice. Six hour after tumor inoculated, the mice were randomly divided into 3 groups.Shenqi (paclitaxel plus Shenqi) or paclitaxel was intraperitoneally injected in two group since the second day of the establishment of animal model. The third group simply administered with normal saline was set as placebo-control. Tumor volume, quantitation of microvessel density (MVD) in inoculated tumor, the number of metastasis in the lungs and survival analysis were compared in 3 groups. RESULTS: Paclitaxel plus Shenqi can effectively reduced MVD in inoculated tumor and the number of lung metastasis as compared with other two group (P<0.05). The survival time of Shenqi group was also significantly longer (P<0.05). Tumor volume was no statistical difference in three group (P>0.05). CONCLUSION: Shenqi can amplify the paclitaxel effect of anti-angiogenesis and anti-metastasis, enhances the survival time of mice bearing LLC, might has possible therapeutic applications in the treatment of lung cancer. PMID- 18394351 TI - Vitamin D, yes, but don't forget iron. PMID- 18394353 TI - Drugs help pass more ureteral stones. PMID- 18394354 TI - Picking a PPI: it comes down to cost. PMID- 18394355 TI - Helping patients kick the "other" habit. PMID- 18394356 TI - Measles hits home: sobering lessons from 2 outbreaks. PMID- 18394357 TI - Bilateral axillary pustules. PMID- 18394358 TI - Lumbar spinal stenosis: can positional therapy alleviate pain? AB - METHODS: We analyzed a retrospective case series of 52 patients with spinal stenosis confirmed by spinal imaging and walking limitations treated with a wheeled walker set to induce lumbosacral flexion. RESULTS: Of the 52 patients, improvement in ambulation was classified as excellent for 30 (58%), good for 7 (13%), moderate for 8 (16%), and poor for 7 (13%). Among 48 patients with neurogenic pain, pain relief was classified as excellent for 22 (46%), good for 11 (23%), moderate for 7 (14.5%), and poor for 8 (16.5%). CONCLUSIONS: These retrospective data from a case series support the hypothesis that positional therapy with a wheeled walker set to induce lumbosacral flexion relieves lower extremity symptoms of spinal stenosis. However, an adequate test of this hypothesis will require randomized trials of sufficient size and duration that include objective clinical endpoints such as quality-of-life measures, immobility complications and need for drugs, physical therapy, procedures including epidural injections, and spinal surgery. In the meantime, this conservative strategy is an option for patients following the recommendations of the North American Spine Society, or for those who have contraindications (or aversions) to surgery or epidural injections, or who have found these options ineffective. Positional therapy with a wheeled walker offers the possibility of short-term benefits for ambulation and pain, with minimal risks and costs. PMID- 18394359 TI - Clinical inquiries. What's the best way to treat Achilles tendonopathy? PMID- 18394360 TI - Clinical inquiries. What is the clinical workup for failure to thrive? PMID- 18394361 TI - Clinical inquiries. How should you evaluate elevated calcium in an asymptomatic patient? PMID- 18394362 TI - Clinical inquiries. What's the most effective treatment for giardiasis? PMID- 18394363 TI - [Lack of association between the APOE genotype and the response to statin treatment in patients with acute ischemic episodes]. AB - BACKGROUND AND OBJECTIVE: APOE genotype has been shown to have an influence on lipid concentrations. However, its relation with response to lipid-lowering treatment is not well established. The aim of our work was to analyze whether this genotype is associated with changes in the lipid profile in response to statins treatment. PATIENTS AND METHOD: A total of 222 consecutive patients with acute ischemic episodes and subjected to treatment with statins were included in a retrospective study. The patients' lipid profile was determined at the first visit to the Lipids Unit and after one year on a statin regime. APOE genotypes were determined by PCR-RFLP, and separated in three groups: E2 (E2 carriers), E4 (E4 carriers) and E3 (E3/E3). E2/E4 patients were not included in the study. RESULTS: Relative frequencies of alleles epsilon2, epsilon3 and epsilon4 were 10.5%, 70.9% and 18.6% respectively. Significant differences among groups (p = 0.039) were observed for c-LDL concentrations. E2 group had lower c-LDL than E3 group (p = 0.017) and E4 group (p = 0.01). No significant differences in c-LDL, c HDL and c-HDL/CT were observed among the three groups with regard to variation after statin treatment. CONCLUSION: APOE genotype does not significantly affect the lipid response in patients with acute ischemic episodes after statin treatment. PMID- 18394364 TI - [Role of tobacco use on the incidence of depression in the SUN cohort study]. AB - BACKGROUND AND OBJECTIVE: Several investigations, most of them cross-sectional, have analyzed the association between smoking and depression, but there are not enough evidences to establish the direction of the association. Our objective was to prospectively assess the association between smoking habit and depression incidence in the SUN (Seguimiento Universidad de Navarra) cohort. SUBJECTS AND METHOD: Dynamic cohort study (recruitment permanently open) based on 8,556 graduate participants where depression incidence was ascertained during 6 years of follow-up. Depression was defined as the presence of a self-reported physician diagnosis and/or use of antidepressant medication in at least one of the 2-year, 4-year or 6-year follow-up questionnaires. The association between smoking and depression incidence was estimated through the calculation of hazard ratios (HR) and their 95% confidence intervals (CI) using Cox regression models (proportional hazards). RESULTS: The mean follow-up period was 47.4 months. One hundred and ninety participants initially free of depression reported a physician diagnosis of depression during follow-up. A significantly higher risk was found for smokers (HR = 1.39; 95% CI, 1.00-1.93) when they were compared to non smokers, whereas an inverse association was found for ex-smokers who had quit smoking more than 10 years ago (HR = 0.42; 95% CI, 0.19-0.94). CONCLUSIONS: This study supports, with a prospective design, an increased risk of depression associated to smoking. PMID- 18394366 TI - [Polymorphisms of apoprotein E and response to statins]. PMID- 18394365 TI - [Screening for high blood pressure among hypertensive and metabolic syndrome patients' children]. AB - BACKGROUND AND OBJECTIVE: Essential arterial hypertension (HTA) and metabolic syndrome (MS) are highly prevalent disorders, with familiar aggregation and important mortality. Late HTA diagnosis is made in a high percentage of people. The aim of this study was to diagnose HTA among hypertensive and MS patients' children. PATIENTS AND METHOD: Hypertensive and MS patients attended at primary and hypertension clinics were studied. Hypertensive patients collaborated to get their children's blood pressure measured. If high blood pressure was detected, the patients' children were referred to their family doctor in order to confirm the diagnosis of HTA. RESULTS: 118 hypertensive patients were studied. Blood pressure was measured in 117 hypertensive patients' children. Thirty-two of them (27.4%) had blood pressure > or = 130/85 mmHg. Eventually, 20 of them (62.5%) were diagnosed of HTA. 17% of the hypertensive patients' children studied were newly diagnosed of HTA. CONCLUSIONS: Screening for HTA among hypertensive and MS patients' children is useful for early diagnosis or HTA and it is cost-effective. PMID- 18394367 TI - [Mediastinic staging of lung cancer in the 21st century: a multidisciplinary challenge]. PMID- 18394368 TI - [Esophageal cancer]. PMID- 18394369 TI - [Hereditary systemic autoinflammatory diseases. Part II: cryopyrin-associated periodic syndromes, pediatric systemic granulomatosis and PAPA syndrome]. AB - Hereditary systemic autoinflammatory diseases result from a genetically-based dysregulated inflammatory process, and are clinically characterized by recurrent or persistent systemic inflammatory episodes, which typically occur in the absence of infectious, neoplastic or autoimmune etiology. Elucidation of their molecular basis has enabled the use of genetic analyses to achieve an accurate and definitive diagnosis, and to establish a tailored treatment. The present review is the second and last part of an updated and comprehensive overview of hereditary systemic autoinflammatory diseases, and will introduce persistent, non periodic autoinflammatory diseases, such as: a) the group of cryopyrin-associated periodic syndromes (CAPS), which includes familial cold-induced autoinflammatory syndrome (FCAS), Muckle-Wells syndrome, and CINCA-NOMID syndrome; b) the group of pediatric systemic granulomatosis, which includes both Blau syndrome and early onset sarcoidosis, and c) the pyogenic sterile arthritis, pyoderma gangrenosum and acne (PAPA) syndrome. PMID- 18394370 TI - [Prospective study of subcutaneous ceftriaxone in patients on palliative care]. PMID- 18394371 TI - [Hepatocellular carcinoma with soft tissue tumour as initial symptom: a case report]. PMID- 18394372 TI - [Polydactylia]. PMID- 18394373 TI - [Practical handling of the asthma based on the control of the disease: update of GINA guidelines]. PMID- 18394374 TI - [Developmental outcome of extremely low birth weight infants (<1,000 g) during the first three years of life]. AB - INTRODUCTION: Obstetrics and Perinatal Care has improved in the last few years. As a result there has been a significant increase in preterm babies and longer survival. The risk of abnormalities in their development is high; they need follow up, prompt action and prevention from the neonatal period. MATERIAL AND METHODS: We evaluated 116 preterm babies, birth weight less than or equal to 1,000 grams, admitted to our Unit over 5 years. Seventy five neonates survived after the neonatal period (64.7 %). We studied the development during the first three years of life. RESULTS: The number of admissions of extremely low birth weigh infants has increased by 100 % in the last five years and the survival has increased by 15 %. Failure to thrive was the most frequent problem; in the first 2 years of life neonatal growth retardation was present in more than 50 % of cases (58 % 1st year, 57 % 2nd year, 40 % 3rd year). Catch-up growth occurred except in infants who had intrauterine growth retardation. Motor function disorders were the most serious; minor alterations in this system were the most frequent but usually did not compromise the life of the patient. Cerebral palsy was present in 14 % in the 1st year of life, 17,4 % 2nd year and 19 % 3rd year. Retinopathy of prematurity was very high in our group (74 %), but ophthalmic sequela were infrequent (3.2 % 1st year, 3.9 % 2nd year, and 3.4 % 3rd year); similarly with hearing function (deafness 2.0 %). Only some mild or moderate language development delay was present and psychomotor development was normal in the majority of cases and those who were retarded improved with time. CONCLUSIONS: Nowadays there is a significant increase in the incidence and survival of extremely low birth weigh infants. They have a higher risk of abnormal development. The most frequent problem is failure to thrive and the most serious is cerebral palsy. Major ophthalmic and hearing sequela are rare. Psychomotor and language development is normal in the majority of these patients. PMID- 18394375 TI - [Influence of birth weight on the lipid profile and blood pressure in Madrid adolescents]. AB - OBJECTIVE: The aim of this study is to assess the relationship between birth weight and the lipid profile and blood pressure in adolescents. PATIENTS AND METHODS: Cross sectional study. We studied seven hundred and forty adolescents aged 18 who attended four different schools located in Madrid. Birth weight (BW) data were obtained from delivery records. All subjects underwent a physical examination, including measurement of blood pressure. Plasma lipids were determined using standardised methods after ten hour fasting. RESULTS: Total cholesterol and cholesterol-LDL in males and females with BW < 3.000 grams were statistically greater than those from subjects with BW > 3,800 grams. Male adolescents also showed a difference in the apolipoprotein B (ApoB) levels. We only found a statistically significant difference in the diastolic blood pressure of male adolescents belonging to the extreme birth weight quintiles. CONCLUSIONS: We found an inverse relationship between birth weight and total cholesterol and cholesterol LDL levels in males and females. This association was also noted in Apo B concentrations in male adolescents. The only influence birth weight had on blood pressure was on the diastolic blood pressure of males. PMID- 18394376 TI - [The effect of helium on ventilator parameters: in vitro study with a Servoi ventilator]. AB - OBJECTIVE: To analyze the changes in respiratory parameters in a Servoi ventilator with heliox. MATERIAL AND METHODS: In vitro study with a Servoi (Maquet) ventilator in volume controlled (VC), pressure controlled (PC) and volume control regulated by pressure (VCPR) modes connected to an artificial lung. A heliox tank with a fixed concentration of helium 70 % and oxygen 30 % was connected to the air inlet of the ventilator. The ventilator was set in VC mode with tidal volumes of 30, 50, 100, 250 and 500 ml; in PC mode with pressure of 20 and 30 cmH(2)O and in VCPR mode with tidal volume of 150 ml. In each case FiO(2) of 21, 30, 40, 50, 60, 70, 80, 90 and 100 % was used. The FiO(2), inspired and expired tidal volume and inspiratory pressure measured by the ventilator and a pitot spirometer (Datex_S5) were compared. RESULTS: In VC and VCPR modes the increase in helium produced a progressive decrease in the pressure needed to administrate the set volume. Heliox also produced a decrease in the tidal volume measured by the spirometer and the tidal expired volume measured by the respirator. In PC mode, heliox produced a progressive increase in the inspired tidal volume, increasing the differences between inspired and expired tidal volumes. CONCLUSIONS: Heliox used with Servoi ventilator produces a decrease in inspiratory pressures in VC and VCPR modes, and an increase in inspiratory tidal volume in PC mode. In all modes heliox reduced the expired tidal volume measured by the ventilator and the spirometer. These changes should be borne in mind if heliox is used with this ventilator. PMID- 18394377 TI - [Use of capnography in acute gastroenteritis]. AB - OBJECTIVE: End-tidal carbon dioxide (PetCO(2)) is a good predictor of circulation and metabolism. There are different studies that suggest PetCO(2) monitoring is a valuable and reliable tool to follow acidosis in pediatric patients with diabetic ketoacidosis. Acute gastroenteritis can cause acidosis, therefore, capnography could be useful in these situations. The objective was to determine the relationship between PetCO(2) and bicarbonate (HCO3) in pediatric patients with acute gastroenteritis and acidosis. METHODS: Clinical, prospective, observational study from April 2006 to January 2007. Children with acute gastroenteritis and dehydration and pH < 7.30 and HCO3 < 20 meq/L in laboratory tests were included. Exclusion criteria included patients with not well tolerated capnography and respiratory illness. Initial and post treatment PetCO(2) and HCO3 were collected as well as demographic data, physical examination data, laboratory tests and hospitalization data. RESULTS: Twenty-five children were included in the final analysis (10 female, 15 male). The mean age was 11.6 + 10 months (1-144 months). The correlation between PetCO(2) and serum bicarbonate was statistically significant with a Pearson's correlation coefficient of r = 0.61 for initial values and r = 0.75 for post treatment values. CONCLUSIONS: Capnography offers a noninvasive measurement of acidosis in pediatric patients with acute gastroenteritis and dehydration. PMID- 18394379 TI - [Congenital amegakaryocytic thrombocytopenia in a 12 year old boy with no signs of pancytopenia: molecular analysis]. AB - Congenital amegakaryocytic thrombocytopenia is an uncommon disorder characterized by an isolated thrombocytopenia due to ineffective megakaryocytopoiesis at birth and the almost complete absence of megakaryocytes in the bone marrow. Mutations in the gene for the thrompoietin receptor MPL were defined as the molecular cause in congenital amegakaryocytic thrombocytopenia patients, developing into a pancytopenia during the first years of life. A case of congenital amegakaryocytic thrombocytopenia with no signs of pancytopenia at the age of 12 years is presented. Mutations were not found in the MPL gene. PMID- 18394378 TI - [End-of-life care in a Spanish Pediatric Intensive Care Unit: staff and parental evaluation]. AB - OBJECTIVES: To evaluate end-of-life care in a Paediatric Intensive Care Unit (PICU). METHODS: Retrospective study developed in a PICU. SUBJECTS: 41 workers from the PICU and parents of 26 deceased children (from 2001 to 2005). A questionnaire was designed to investigate end-of-life care. RESULTS: An age < 1 year old and a chronic or congenital disease correlated with a perceived lack of consistency in information. Nearly 38 % parents were with their children at the time of death; 64 % of all parents consider this "positive", and 13 % consider it "negative". Forty per cent of staff stated that it is "positive" for parents to be by the side of their child at the time of death, and 52 % do not know. Seventy three per cent of staff, but only 29 % of parents want further professional psychological support for parents. Twenty per cent of children died following withdrawal of life support. The most important factors for this decision were the possibility of survival and quality of life. The majority (73 %) of caregivers express the view that often, this decision should be taken earlier. CONCLUSIONS: Analysis of staff opinions underlines the importance of the way news is communicated, the timing of withdrawal of life support, and the need for psychological support. Parents emphasized the role of the family during time spent in a PICU and during the last moments. PMID- 18394380 TI - [Persistent pulmonary hypertension of the newborn following ingestion of nonsteroidal anti-inflammatory drugs during pregnancy]. AB - Nonsteroidal anti-inflammatory drugs induce the inhibition of prostaglandin synthesis, which can cause constriction of the fetal ductus arteriosus in the pregnancy. We report two cases of antenatal closure of ductus arteriosus with severe pulmonary hypertension following maternal ingestion of nonsteroidal anti inflammatory drugs (niflumic acid and acetylsalicylic acid) in the last days before delivery. To our knowledge, this is only the second case in literature describing antenatal closure of ductus arteriosus after the administration of niflumic acid. Prescription of nonsteroidal anti-inflammatory drugs must be avoided during pregnancy. Fetal echocardiography must be monitored in those women treated with nonsteroidal anti-inflammatory drugs. PMID- 18394381 TI - [Partial duplication of chromosome 4q (q31, q35): Auriculo-acro-renal syndrome]. AB - The partial trisomy 4q is a strange chromosomal illness. This illness is caused by the duplication of a portion of chromosome 4. In most of the cases, it is the result of a balanced translocation in one of the progenitors. The "de novo" appearance is less common. We present a patient with a partial "de novo" duplication in the distal segment of the long arm of chromosome 4 (q31, q35), in association with Robertsonian translocation between chromosomes 14 and 21. This association has not been described previously. In the 4q duplication, the relationship between the phenotype and the parts of the duplicated segment is not well defined, although it seems clear that the renal anomalies and/or thumbs abnormalities are a characteristic manifestation. We have reviewed the literature and, of the cases previously described with trisomy q31-35, we came to the conclusion that this region of chromosome 4 may also be involved in constituting the "Syndrome of partial trisomy 4q" or Auriculo-acro-renal Syndrome". PMID- 18394382 TI - [SOS1 mutation: a new cause of Noonan syndrome]. AB - Noonan syndrome, characterized by short stature, facial anomalies, heart disease and cryptorchidism in males, is an autosomal dominant, genetically heterogeneous disease. Approximately 50 % of Noonan syndrome cases are caused by gain-of function mutations in PTPN11, encoding the tyrosine phosphatase (SHP2) and 5 % are caused by KRAS mutations. Recently, a new mutation in SOS1 gene has been identified in approximately 20 % of cases of Noonan syndrome without PTPN11 mutation. That difference in genotype has a relationship with phenotype that we must investigate. We report a case of Noonan syndrome due to an SOS1 mutation; we describe his phenotype and subsequent outcome. PMID- 18394383 TI - [Peutz-Jeghers syndrome]. AB - Peutz-Jeghers syndrome is an inherited disorder which usually debuts during childhood. It is characterized by mucocutaneous pigmentation and hamartomatous polyps in the gastrointestinal tract. Numerous reports indicate a high incidence of gastrointestinal and extraintestinal cancer in these patients, their appearance at a young age, as well as its association with ovarian and testicular tumors. An aggressive approach of these patients seems to be necessary. We report the case of two brothers suffering from Peutz-Jeghers syndrome whose father and grandfather died as a consequence of the progression of an intestinal cancer related to the syndrome. PMID- 18394384 TI - [Community-acquired, methicillin-resistant Staphylococcus aureus acute osteomyelitis and pneumonia]. AB - The appearance of a community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is a increasingly important problem. We describe the case of a child suffering a CA-MRSA osteomyelitis complicated with a non-necrotizing pneumonia. The fast resolution of the pulmonary infection contrasts with the poor evolution of the septic bone. We emphasise the importance of correct surgical drainage to clear the infection. We review the literature on this emergent pathogen. PMID- 18394385 TI - [Risks factors for pediatric malignant liver tumors]. AB - OBJECTIVE: Pediatric Hepatic Malignancies (PHMs) are the result of the interaction between constitutional and environmental risk factors (RFs). We review the evidence on the main RFs associated to PHMs. METHOD: Systematic review of the literature published in the last 25 years on Medline, Embase, Cancerlit, Lilacs and SciElo using the following key words: "etiology/risk factor/epidemiology" and "malignant liver tumors/hepatic cancer" or "hepatoblastoma/hepatocarcionoma". RESULTS: PHMs account for 1 % of all pediatric malignancies. The main types, hepatoblastoma (HB) and hepatocarcionma (HCC) make up 98-99 % of PHM. The main constitutional RFs are: a) Beckwith-Wiedemann (BW) syndrome; b) isolated hemihyperplasia syndrome (IHS); c) adenomatous polyps of the colon; d) hemochromatosis; e) Hereditary Tyrosinemia Type 1; f) a -1 antitrypsin deficiency; g) porphyrias; h) cirrhosis; i) nonalcoholic steatosis; and j) primary sclerosing cholangitis. The main environmental RFs are: a) hepatitis B virus (HBV) and C virus (HCV); b) B1 aflatoxin (B1AF); c) ionizing radiation; d) alcohol; e) hormonal treatments; f) occupational exposure to pesticides, solvents, vinyl chloride and metals; g) smoking; h) arsenic; i) prematury and very low birth weight; and j) trematodes. CONCLUSIONS: The clinical, analytical and ultrasound screening facilitate the early diagnosis of HB in the previously mentioned genetic syndromes, particularly BW and IHS during the first years of life. HBV universal vaccination of newborns provides the biggest opportunity to prevent a substantial proportion of PHMs. Also systematic monitoring of HBV and HCV in blood, hemoderivates, donated organs and drug addicts, are very useful. Other effective measures are: the reduction/elimination of B(1)AF in food, zero alcohol intake during childhood and adolescence as well decreasing prenatal exposure to the tobacco, solvents, pesticides, vinyl chloride, metals, ionizing radiation and hormonal treatments. PMID- 18394386 TI - [Use of statins in children]. AB - In the last decade treatment of hyperlipidemia has changed substantially due to the use of statins, a group of drugs that inhibit the enzyme hydroxymethylglutaryl coenzyme-A-reductase (HMG-CoA reductase). Its use has been proven to be efficient and safe in adults, and could also be of benefit in the treatment of hypercholesterolemia in children. Up to date, six of these drugs have been used in children: lovastatin, fluvastatin, pravastatin, simvastatin, atorvastatin and more recently, rosuvastatin. In some of these studies the follow up has been up to three years and, as regards their use during childhood, we can conclude that they are safe as well as efficient in reducing the blood levels of total cholesterol (TC) and low-density lipoproteins cholesterol (LDL-C). PMID- 18394387 TI - [Glossary of bioethic terms and expressions frequently used in pediatric intensive care practice]. AB - Ethical issues are of increasing interest in current medicine, and pediatrics is no exception. In critical care, the relevance of these considerations becomes even greater. Commonly used expressions in bioethics, frequently lead to terminological confusion and misunderstandings, as reported by several publications, revealing a lack of clear concepts in many cases. As an attempt to clarify or facilitate the comprehension of the most relevant terms in this field, the Spanish Society of Pediatric Intensive Care has prepared a Glossary of the most commonly used terms and expressions. PMID- 18394388 TI - [Congenital cerebral hemiatrophy]. PMID- 18394389 TI - [Absence of the greater sphenoid wing: an exceptional etiology of pulsatory exophthalmos]. PMID- 18394390 TI - [X- linked adrenoleukodystrophy: misdiagnosed disease in children with idiopathic Addison's disease]. PMID- 18394391 TI - [Is the CRIB a good predictor of intraventricular hemorrhage?]. PMID- 18394393 TI - [Selective neonatal screening for sickle cell disease]. PMID- 18394394 TI - [Hereditary hyperferritinemia and cataract syndrome: a de novo mutation]. PMID- 18394395 TI - [Dacryocystocele with nasal mucocele causing neonatal respiratory distress]. PMID- 18394396 TI - [Facial dermatophytide reaction]. PMID- 18394397 TI - [Persistent vena hemiazygos and absence of intrahepatic inferior vena cava. A casual finding?]. PMID- 18394398 TI - [The social structure of dermatology: between the medieval and the early modern]. PMID- 18394399 TI - [Rosacea]. AB - Rosacea is a chronic inflammatory skin disease appearing in the central area of the face of middle-aged patients. It is characterized by flushing, permanent erythema, telangiectasia, papules, pustules, and the absence of comedones. Its underlying pathophysiological mechanisms are not completely understood, although a number of hypotheses point to vascular abnormalities and infection by microorganisms such as Demodex folliculorum. Rosacea is classified into 4 subtypes, which determine the therapeutic approach based on skin care, topical antiinflammatory agents, topical and oral antibiotics and retinoids, and, in some instances, light-based therapy and surgery. PMID- 18394400 TI - [Management of androgenetic alopecia in postmenopausal women]. AB - Female androgenetic alopecia or female-pattern alopecia is one of the most common causes of hair loss, affecting 50 % of women over their lifetime. The appearance of this condition is the cause of significant stress and psychological problems, making appropriate management important. Cases exist in which it is associated with hyperandrogenism. Here, we review the different clinical forms (diffuse, male-pattern, and Christmas-tree pattern), discuss the most appropriate laboratory tests (complete blood count, thyroid stimulating hormone, ferritin, prolactin, free and/or total testosterone, and dehydroepiandrosterone sulfate), and the different treatments, including finasteride. PMID- 18394401 TI - [Recent advances in laser therapy and other technologies]. AB - Laser technology and other energy sources are rapidly finding a place in dermatology clinics. In the field of skin rejuvenation by fractional photothermolysis, although few controlled studies have been undertaken, several devices have emerged in recent years that appear less effective than laser ablation techniques but that are safer. The aim of this short article is to provide an introduction, though not treat in depth, the different emerging technologies in dermatology. We will focus particularly on lasers and light sources in improving applications such as the treatment of vascular lesions, acne, and encapsulated ink tattoos; light-emitting diodes; developments in the treatment of cellulitis; photodynamic therapy; suction methods; scarring; and finally the recent and doubtlessly future introduction of home devices for use in a range of dermatologic applications (depilation, rejuvenation, treatment of acne, etc). PMID- 18394402 TI - [Epidermodysplasia verruciformis-like cells as histologic markers of immunosuppression: review of 229 squamous cell carcinomas]. AB - INTRODUCTION: Epidermodysplasia verruciformis (EV) is associated with greater susceptibility to infection by certain oncogenic subtypes of human papillomavirus (HPV). Among other histologic findings, large, clear, oval or rounded cells (EV cells) are observed in the granular layer in EV, and some authors consider these cells to be markers of immunosuppression. MATERIAL AND METHODS: We analyzed 229 squamous cell carcinomas (SCC) to determine whether EV cells were present and to assess whether their presence was associated either with localized or cutaneous immunosuppression (tumors with signs of severe chronic actinic damage or severe stasis dermatitis) or with systemic immunosuppression (immunocompromised or elderly patients). RESULTS: We observed EV cells in 33 SCC. No statistically significant relationship was observed between the presence of EV cells and immunosuppression. We performed polymerase chain reaction in 8 lesions, but the results were not informative as the DNA was denatured. CONCLUSIONS: We found no relationship between the presence of EV cells and localized or systemic immunosuppression, possibly because the sample was inadequate (almost all SCC studied were associated with signs of immunosuppression, irrespective of the presence or absence of EV cells). Further studies will be required to compare lesions associated with immunosuppression with those in which immunosuppression is absent. The presence of EV cells may be the result of cytopathic effects of certain HPV subtypes, such as HPV 5 or 8, but this will need to be demonstrated using techniques such as polymerase chain reaction. PMID- 18394403 TI - [Mohs micrographic surgery: our first 100 patients]. AB - INTRODUCTION: Mohs surgery was first described by Frederic Mohs in 1941 to eliminate high-risk cutaneous tumors. The technique involves histological examination of the margins. OBJECTIVE: The aim of this study was to describe the clinical findings and compare the grade and subclinical extension of the tumor as measured by the number of Mohs stages needed for complete elimination according to whether the tumor presented high-risk factors. METHODS: We included 100 patients with 105 tumors. In all cases, age, sex, tumor site, tumor type, histological subtype in the case of basal cell carcinoma, size, recurrences, number of Mohs stages, and reconstruction technique were recorded. RESULTS: The study group comprised 44 men and 56 women aged between 28 and 88 years (mean, 72.6 years). Of the tumors, 80 % corresponded to basal cell carcinoma, 12.38 % to squamous cell carcinoma, 4.76 % to dermatofibrosarcoma protuberans, 0.95 % to Merkel cell carcinoma, 0.95 % to microcystic adnexal carcinoma, and 0.95 % to lentigo maligna melanoma. Most tumors were located on the head and 60 % required more than one Mohs stage for complete elimination. CONCLUSION: This series is characterized by a high percentage of high-risk cutaneous tumors. After assessing the risk factors independently, we found that the size of the tumor is the risk factor most closely related to grade and subclinical extension in the case of basal cell carcinoma, although similar conclusions cannot be drawn for the other types of tumor studied. PMID- 18394404 TI - [Chemotherapy-induced acral erythema: a clinical and histopathologic study of 44 cases]. AB - INTRODUCTION: Acral erythema, also known as palmoplantar erythrodysesthesia or hand-foot syndrome, is a relatively common cutaneous reaction caused by a variety of chemotherapeutic agents. It presents during cancer treatment as painful erythema and paresthesia affecting the palms and soles. It seems to be dose dependent and its appearance is determined by both the peak plasma concentration and the cumulative dose of the chemotherapeutic agent. The symptoms and histopathology findings are suggestive of direct cytotoxicity affecting the epidermis of the extremities caused by high concentrations of chemotherapeutic agents. The most commonly implicated agents are doxorubicin, 5-fluoracil and its derivatives, cytarabine, and docetaxel. MATERIAL AND METHODS: We present the clinical and histologic characteristics of a series of patients diagnosed with chemotherapy-induced acral erythema. The study included all patients who developed acral erythema lesions following chemotherapy between January 2000 and December 2003. RESULTS AND CONCLUSIONS: Out of 2186 patients who underwent chemotherapy, 44 cases of acral erythema were identified, representing an incidence of 2.01 % during the study period and 16.75 % of all cutaneous lesions attributed to chemotherapy. The most commonly implicated drug was 5-fluoracil administered by continuous infusion and the highest incidence was observed in patients treated with liposomal doxorubicin. Acral erythema was a dose-limiting toxic effect in 29.5 % of cases. The histologic findings varied according to the clinical severity of the lesions and included interface dermatitis with variable keratinocyte necrosis, dilation of the superficial vascular plexus, and limited inflammatory infiltrate. The most commonly used treatment was pyridoxine, along with topical treatments such as cold compresses, emollients, and topical corticosteroids. PMID- 18394405 TI - [Cutaneous follicular center B-cell lymphoma treated with intralesional rituximab]. AB - Cutaneous follicular center B-cell lymphomas are indolent tumors characterized by the presence of neoplastic follicular center cells. They contain a mixture of centrocytes with a variable number of centroblasts. The tumor is usually treated by surgery or radiotherapy, although other treatments may be used such as interferon-alpha, chemotherapy, and biological agents (rituximab). Rituximab is a chimeric monoclonal anti-CD20 antibody that can be administered intravenously or intralesionally. We report the case of a 41-year-old man who consulted for violaceous nodular lesions in the left scapular region and who was diagnosed with cutaneous follicular center B-cell lymphoma after biopsy, laboratory tests, thoracic-abdominal-pelvic computed tomography, abdominal ultrasound, and bone marrow biopsy. It was decided to treat him with 30 mg of intralesional rituximab administered for 1 week (3 times) every month for 4 months. Complete response was obtained. We also review the published cases of cutaneous B-cell lymphoma treated with intralesional rituximab. PMID- 18394406 TI - [A case of cutaneous extramedullary hematopoiesis associated with idiopathic myelofibrosis]. AB - Cutaneous extramedullary hematopoiesis is a rare manifestation of chronic myeloproliferative processes, mainly chronic idiopathic myelofibrosis. In adults, it manifests as macules, papules, nodules, and ulcers on the trunk. The lesions usually appear soon after diagnosis and the possibility of a relationship between splenectomy and the appearance of extramedullary foci of hematopoiesis is still debated. Diagnosis is based on histopathology showing an infiltrate with different combinations of myeloid and erythroid cell precursors and megakaryocytes. Symptomatic treatment is provided alongside treatment of the underlying disease. We report a new case associated with chronic idiopathic myelofibrosis in which foci of cutaneous extramedullary hematopoiesis were observed 9 years after initial diagnosis. The lesions were progressive and the patient went on to develop acute myeloid leukemia. PMID- 18394407 TI - [Six cases of nevus oligemicus]. AB - Hamartoma or nevus oligemicus is an uncommon lesion that is characterized by selective vasoconstriction of the deep dermal vascular plexus with respect to the superficial one and whose cause has not been clearly established. This selective vasoconstriction gives rise to fixed, acquired, and asymptomatic lesions in the form of livid, erythematous macules that are typically cold to touch compared with surrounding skin. We report the cases of 6 young men with lesions clinically compatible with nevus oligemicus on the abdomen and flanks. Measurement of the surface temperature of the lesion revealed a decrease of up to 2.5 degrees C with respect to healthy surrounding skin and allowed a definitive diagnosis to be made. We describe the additional studies undertaken, the differential diagnosis, and the possible etiologic agents, and discuss the cases reported in the literature to date. In our opinion, nevus oligemicus is an underdiagnosed lesion that is much more common than has been reported in the literature. PMID- 18394408 TI - [Congenital tumor with a vascular appearance]. PMID- 18394409 TI - [Unusual vascular pattern of a nodular lesion in the dermoscopic examination of a kidney transplant recipient]. PMID- 18394410 TI - [Cutaneous vegetative pseudoepithelioma]. PMID- 18394411 TI - [Cycling: more than a hobby]. PMID- 18394412 TI - [Sturge-Weber syndrome and type 1 neurofibromatosis: a chance association?]. PMID- 18394413 TI - [Essential progressive telangiectasia]. PMID- 18394414 TI - [Extranodal natural killer/T-cell lymphoma, nasal type]. PMID- 18394415 TI - [Paraneoplastic ichthyosis]. PMID- 18394416 TI - [Erythema nodosum associated with inflammatory tinea capitis (kerion celsi)]. PMID- 18394417 TI - [Indapamide-associated Stevens-Johnson syndrome]. PMID- 18394418 TI - Chemiluminescent detection systems of horseradish peroxidase employing nucleophilic acylation catalysts. AB - The light output of the peroxidase-catalyzed luminol chemiluminescent oxidation reaction can be greatly increased by incorporating different enhancers. Such an increase is attributed to the preferential oxidation of the enhancer by peroxidase intermediates and the rapid formation of enhancer radicals that, in turn, quickly oxidize luminol to its radical anion. These enhancers, which include substituted phenols, substituted boronic acids, indophenols, and N-alkyl phenothiazines, behave as electron transfer mediators. A further, very significant increase in light output was also observed by the addition of nucleophilic acylation catalyst to the enhancer/luminol/oxidant substrate. The effect of the new component is general and applicable to many of the known enhancers but is much more remarkable in association with phenothiazine enhancers (up to 10-fold light output). The addition of a nucleophilic acylation catalyst to these substrates lowered the limit of detection for horseradish peroxidase from 50 to 8 amol. Similar improvements were observed in "sandwich" enzyme-linked immunosorbent assays and Western blot assays. PMID- 18394419 TI - Tracking single quantum dot and its spectrum in free solution with controllable thermal diffusion suppression. AB - Thermal motions of semiconductor quantum dots (QDs) are suppressed on a dehydrated agarose-modified surface. The diffusion coefficients (D) of particles can be controlled by modifying the surface with an appropriate agarose concentration. The value of D is more than 100 times lower than the theoretical value when the dried agarose surface is made with an 8% agarose solution. This makes it possible to real-time record the diffusion process of single particles and single molecules in low-viscosity solution. A transmission grating installed in front of the charge-coupled device separates the QD fluorescence into the zeroth-order and first-order spectrum. Therefore, the spectrum of dynamic QDs is tracked on the modified surface. Tracking the dynamic QD spectral image is a promising method to explore the process of the molecular interactions in the physiological buffer. PMID- 18394420 TI - Identification of the RNA N-glycosidase activity of ricin in castor bean extracts by an electrochemiluminescence-based assay. AB - A simple electrochemiluminescence-based assay for RNA N-glycosidase activity has been modified to permit its use with authentic extracts of Ricinus communis (castor beans) and Abrus precatorius (jequirity seeds)--the natural sources of ricin and abrin. Modifications include the addition of an RNase inactivator to the reaction mixture, elimination of a signal-enhancing monoclonal antibody, and optimization of the incubation temperature. Concurrent testing with two substrates provides a diagnostic tool enabling castor bean toxins to be differentiated from a larger selection of N-glycosidase toxins than was previously examined. PMID- 18394421 TI - Peroxynitrite induced formation of the neurotoxins 5-S-cysteinyl-dopamine and DHBT-1: implications for Parkinson's disease and protection by polyphenols. AB - Mechanisms of nigral cell injury in Parkinson's disease remain unclear, although a combination of increased oxidative stress, the formation of catecholamine quinones and the subsequent formation of neurotoxic cysteinyl-catecholamine conjugates may contribute. In the present study, peroxynitrite was observed to generate both 2-S- and 5-S-cysteinyl-dopamine and a dihydrobenzothiazine species, DHBT-1, following the reaction of dopamine with l-cysteine. The formation of 5-S cysteinyl-dopamine and DHBT-1 in the presence of peroxynitrite induced significant neuronal injury. Pre-treatment of cortical neurons with pelargonidin, quercetin, hesperetin, caffeic acid, the 4'-O-Me derivatives of catechin and epicatechin (0.1-3.0 microM) resulted in concentration dependant protection against 5-S-cysteinyl-dopamine-induced neurotoxicity. These data suggest that polyphenols may protect against neuronal injury induced by endogenous neurotoxins relevant to the aetiology of the Parkinson disease. PMID- 18394423 TI - Assembly of the Escherichia coli NADH:ubiquinone oxidoreductase (complex I). AB - The proton-pumping NADH:ubiquinone oxidoreductase is the first of the respiratory chain complexes in many bacteria and the mitochondria of most eukaryotes. In general, the bacterial complex consists of 14 different subunits. In addition to the homologues of these subunits, the mitochondrial complex contains approximately 31 additional proteins. While it was shown that the mitochondrial complex is assembled from distinct intermediates, nothing is known about the assembly of the bacterial complex. We used Escherichia coli mutants, in which the nuo-genes coding the subunits of complex I were individually disrupted by an insertion of a resistance cartridge to determine whether they are required for the assembly of a functional complex I. No complex I-mediated enzyme activity was detectable in the mutant membranes and it was not possible to extract a structurally intact complex I from the mutant membranes. However, the subunits and the cofactors of the soluble NADH dehydrogenase fragment of the complex were detected in the cytoplasm of some of the nuo-mutants. It is discussed whether this fragment represents an assembly intermediate. In addition, a membrane-bound fragment exhibiting NADH/ferricyanide oxidoreductase activity and containing the iron-sulfur cluster N2 was detected in one mutant. PMID- 18394422 TI - Articular cartilage tensile integrity: modulation by matrix depletion is maturation-dependent. AB - Articular cartilage function depends on the molecular composition and structure of its extracellular matrix (ECM). The collagen network (CN) provides cartilage with tensile integrity, but must also remodel during growth. Such remodeling may depend on matrix molecules interacting with the CN to modulate the tensile behavior of cartilage. The objective of this study was to determine the effects of increasingly selective matrix depletion on tensile properties of immature and mature articular cartilage, and thereby establish a framework for identifying molecules involved in CN remodeling. Depletion of immature cartilage with guanidine, chondroitinase ABC, chondroitinase AC, and Streptomyces hyaluronidase markedly increased tensile integrity, while the integrity of mature cartilage remained unaltered after depletion with guanidine. The enhanced tensile integrity after matrix depletion suggests that certain ECM components of immature matrix serve to inhibit CN interactions and may act as modulators of physiological alterations of cartilage geometry and tensile properties during growth/maturation. PMID- 18394424 TI - Short-term down-regulation of zeaxanthin epoxidation in Arabidopsis thaliana in response to photo-oxidative stress conditions. AB - The epoxidation of zeaxanthin (Zx) to violaxanthin after exposure to different light stress conditions has been studied in Arabidopsis (Arabidopsis thaliana). Formation of Zx was induced by illumination of intact leaves for up to 8 h at different light intensities and temperatures. The kinetics of epoxidation was found to be gradually retarded with increasing light stress during pre illumination, indicating a gradual down-regulation of the Zx epoxidase activity. Retardation of the epoxidation rates by a factor of up to 10 was inducible either by increasing the light intensity or by extending the illumination time or by decreasing the temperature during pre-illumination. The retardation of the epoxidation kinetics was correlated with a decrease of the PSII quantum efficiency after the pre-illumination treatment. Experiments with the stn7/stn8 mutant of Arabidopsis indicated that the thylakoid protein kinases STN7 and STN8, which are required for the phosphorylation of PSII proteins, are not involved in the short-term down-regulation of Zx epoxidation. However, the retardation of Zx epoxidation was maintained in thylakoids isolated from pre-illuminated leaves, indicating that a direct modification of the Zx epoxidase is most likely involved in the light-induced down-regulation. PMID- 18394425 TI - Controlled release mechanisms of spontaneously forming unilamellar vesicles. AB - Spontaneously forming small unilamellar vesicles (SULVs) are easy to prepare and show great promise for use in delivering therapeutic payloads. We report of SULVs made up of the ternary phospholipid mixture, dimyristoyl-phosphatidylcholine (DMPC), dihexanoyl-phosphatidylcholine (DHPC) and dimyristoyl phosphatidylglycerol (DMPG), which have been characterized by small angle neutron scattering (SANS). These low-polydispersity (0.14-0.19) SULVs range in size (i.e., radius) from 110 to 215 A and are capable of entrapping, and subsequently releasing, hydrophilic molecules (e.g., fluorescent dyes and quenchers) in a controlled fashion over two different temperature ranges. The low-temperature release mechanism involves the SULVs transforming into discoidal micelles, with an onset temperature (T(o)) of ~32 degrees C, while the high-temperature release mechanism is more gradual, presumably the result of defects formed through the continuous dissolution of DHPC into solution. Both of these mechanisms differ from other, previously reported thermosensitive liposomes. PMID- 18394426 TI - Thermotropic and barotropic phase transitions of dilauroylphosphatidylcholine bilayer. AB - The bilayer phase transitions of dilauroylphosphatidylcholine (DLPC), containing two linear acyl chains with 12 carbon atoms, were observed by means of differential scanning calorimetry (DSC) under ambient pressure and light transmittance under high pressure. When the heating scan for the DLPC bilayer in 50 wt.% aqueous ethylene glycol (EG) solution began at -30 degrees C after cold storage, the DSC thermogram showed two endothermic peaks at 1.7 and 4.5 degrees C, which correspond to the transition from the lamellar crystalline (Lc) phase to the intermediate liquid crystalline (Lx) phase and the transition from the Lx phase to the liquid crystalline (L) phase, respectively. Extremely large enthalpy change (32.9 kJ mol(-1)) is characteristic of the Lc/Lx phase transition. The DSC thermogram for the heating scan beginning from -10 degrees C showed a single endothermic peak with 9.2 kJ mol(-1) at -0.4 degrees C, which was assigned as the so-called main transition between the metastable ripple gel (P'(beta)) and metastable Lalpha phases. The DLPC bilayer under high pressure underwent three kinds of transitions in EG solution, whereas only one transition was observed in water under high pressure. The middle-temperature transition in EG solution could be assigned to the main transition because of its consistency with the main transition in water. The lower-temperature transition is probably assigned as transition from the Lc phase to the P'(beta) phase. Since the slope (dT/dp) of the Lc/P'(beta) phase boundary is smaller than that for the main transition, the Lc/P'(beta) phase boundary and the main transition curves crossed each other at 40 MPa on the temperature-pressure phase diagram. The higher-temperature transition in EG solution refers to the transition from the Lx phase to the Lalpha phase. The Lx phase disappeared at about 180 MPa, and the direct transition from the P'(beta) phase to the Lalpha phase was observed at high pressures above 180 MPa. PMID- 18394427 TI - Glutathione level and glutathione-dependent enzyme activities in blood serum of patients with gastrointestinal tract tumors. AB - OBJECTIVES: Glutathione (GSH) and enzymes cooperating with it - glutathione peroxidase (GSHPx), glutathione S-transferase (GST) and glutathione reductase (GSHR) - play crucial role in cell defence against reactive oxygen species (ROS), which are implicated in tumor disease. The aim of this study was to determine if neoplastic diseases of gastrointestinal tract may influence blood GSH level and its dependent enzyme activity. DESIGN AND METHODS: Blood serum was obtained before and after surgery from patients with gastric, liver and colorectal cancers, and colorectal cancer liver metastases. Lipid peroxidation and GSH levels, and GSH-dependent enzyme activities were determined by means of spectrophotometric methods. RESULTS: Increased level of lipid peroxidation and significant differences in GSH level and GSHPx, GST and GSHR activities were observed in serum taken before and after surgery from patients with gastrointestinal tract tumors compared to those in control serum (from healthy blood donors). CONCLUSIONS: Increase of lipid peroxidation and changes in GSH level and related enzyme activities, suggest oxidative stress in serum of patients with gastrointestinal tract tumor causes, which probably arise as a result of enormous production of ROS in the system. These alterations reflect the presence of functional defence mechanism against oxidative stress related firmly to the glutathione metabolism. PMID- 18394428 TI - AdipoR1 mediates the anorexigenic and insulin/leptin-like actions of adiponectin in the hypothalamus. AB - Adiponectin exerts an insulin-sensitizing effect, improving insulin action in peripheral tissues and restraining insulin resistance. Here, we explore the hypothesis that adiponectin can reproduce some of the actions of insulin/leptin in the hypothalamus. The presence of AdipoR1 and AdipoR2 was mapped to the arcuate and lateral hypothalamic nuclei. Icv adiponectin reduced food intake, which was accompanied by activation/engagement of IRS1/2, ERK, Akt, FOXO1, JAK2 and STAT3. All these actions were dependent on AdipoR1, since inhibition of this receptor, and not of AdipoR2, completely reversed the effects described above. Thus, adiponectin acts in the hypothalamus, activating elements of the canonical insulin and leptin signaling pathways and promoting reduction of food intake. PMID- 18394429 TI - Regulation of mRNP dynamics along the export pathway. AB - The transcription of mRNA is tightly coupled to the concomitant recruitment of mRNA processing and export factors, resulting in the formation of mature and export competent mRNP complexes. This interconnection in gene expression implies extensive spatio-temporal control of mRNP dynamics to prevent mRNA export factors bound to pre-mRNA from functioning at the incorrect time and exporting nascent or incompletely processed pre-mRNAs. Recent discoveries provide molecular understanding of how a broad range of post-translational modifications together with RNA-dependent ATPases coordinate proteins acting at different steps and regulate mRNP assembly and export. PMID- 18394430 TI - Born to run; the story of the PEPCK-Cmus mouse. AB - In order to study the role of the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK-C) in skeletal muscle, PEPCK-Cmus mice were created by introducing the cDNA for the enzyme, linked to the human alpha skeletal actin gene promoter, into their germ line. Two founder lines generated by this procedure were bred together, creating a line of mice that have 9.0 units/g skeletal muscle of PEPCK-C, as compared to 0.080 units/g in muscle from control animals. The mice were more active than controls in their cages and could run for up to 5 km, at a speed of 20 m/min without stopping (control mice run for 0.2 km at the same speed). Male PEPCK-Cmus mice are extremely aggressive, as well as hyperactive. During strenuous exercise, they use fatty acids as a fuel more efficiently than do controls and produce far less lactate than do control animals, perhaps due to the greatly increased number of mitochondria in their skeletal muscle. PEPCK-Cmus mice also store up to five-times more triglyceride in their skeletal muscle, but have only marginal amounts of triglyceride in their adipose tissue depots, despite eating 60% more than controls. The concentration of leptin and insulin the blood of 8-12 months of PEPCK-Cmus mice is far lower than noted in the blood of control animals of the same age. These mice live longer than controls and the females remain reproductively active for as long as 35 months. The possible reasons for the profound alteration in activity and longevity caused the introduction of a simple metabolic enzyme into the skeletal muscle of the mice will be discussed. PMID- 18394431 TI - Deleterious effects due to glucose or to triose phosphates. PMID- 18394432 TI - Induction of apoptosis by cigarette smoke via ROS-dependent endoplasmic reticulum stress and CCAAT/enhancer-binding protein-homologous protein (CHOP). AB - In this report, we investigated a role of endoplasmic reticulum (ER) stress in cigarette smoke (CS)-induced apoptosis of human bronchial epithelial cells (hBEC). Exposure of hBEC to CS or CS extract (CSE) caused expression of endogenous ER stress markers GRP78 and CHOP and induction of apoptosis evidenced by nuclear condensation, membrane blebbing, and activation of caspase-3 and caspase-4. In vivo exposure of mice to CS also caused induction of GRP78 and CHOP in the lung. Attenuation of ER stress by overexpression of ER chaperone GRP78 or ORP150 significantly attenuated CSE-triggered apoptosis. Exposure of hBEC to CSE caused generation of reactive oxygen species, and treatment with antioxidants inhibited CSE-induced apoptosis. Interestingly, antioxidants including a scavenger of O(2)(*-) blunted induction of CHOP by CSE without affecting the level of GRP78, and dominant-negative inhibition of CHOP abolished CSE-induced apoptosis. Furthermore, a generator of O(2)(*-) selectively induced CHOP and apoptosis in hBEC. Our results revealed that: (1) CS induces ER stress in vitro and in vivo, (2) ER stress mediates CS-triggered apoptosis downstream of oxidative stress, (3) CS-initiated apoptosis is caused through oxidative stress dependent induction of CHOP, (4) O(2)(*-) may play a dominant role in this process, and (5) oxidative stress-independent induction of GRP78 counterbalances the proapoptotic action of CHOP. PMID- 18394433 TI - Immunolocalization and enzymatic functional characterization of the thioredoxin system in Entamoeba histolytica. AB - The components of the redox metabolism in Entamoeba histolytica have been recently revisited by Arias et al. (Free Radic. Biol. Med. 42:1496-1505; 2007), after the identification and characterization of a thioredoxin-linked system. The present work deals with studies performed for a better understanding of the localization and identification of different components of the redox machinery present in the parasite. The gene encoding for amoebic thioredoxin 8 was cloned and the recombinant protein typified as having properties similar to those of thioredoxin 41. The ability of these thioredoxins and the specific reductase to assemble a system utilizing NADPH to metabolize hydroperoxides in association with a peroxiredoxin has been kinetically characterized. The peroxiredoxin behaved as a typical 2 cysteine enzyme, exhibiting a ping-pong mechanism with hyperbolic saturation kinetics for thioredoxin 8 (K(m)=3.8 microM), thioredoxin 41 (K(m)=3.1 microM), and tert-butyl hydroperoxide (K(m) about 35 microM). Moreover, the tandem system involving thioredoxin reductase and either thioredoxin proved to be operative for reducing low molecular weight disulfides, including putative physiological substrates as cystine and oxidized trypanothione. Thioredoxin reductase and thioredoxin 41 (by association also the functional redox system) have been immunolocalized underlying the plasma membrane in Entamoeba histolytica cells. These findings suggest an important role for the metabolic pathway involving thioredoxin as a redox interchanger, which could be critical for the maintenance and virulence of the parasite when exposed to highly toxic reactive oxygen species. PMID- 18394434 TI - Activation properties of GAGA transcription factor. AB - GAGA is a Drosophila transcription factor that has been involved in many nuclear activities. We present evidence that GAGA factor enhances transcription by stabilizing pre-initiation complex (PIC) and promoting reinitiation. Formation of PIC prior to GAGA addition prevents activation suggesting that GAGA is required early in the formation of activated complexes. GAGA stimulation of transcription can be attributed in part to a stabilization of PIC. All these properties depend on the GAGA C-terminal glutamine-rich domain and, in addition to other roles and together with previous data, support a role of GAGA as a transcription factor. PMID- 18394435 TI - Baseline plasma adiponectin levels as a predictor of left ventricular systolic dysfunction in patients referred for coronary angiography. AB - The objective of the present study was to determine the association between plasma adiponectin and left ventricular (LV) systolic function. Baseline plasma adiponectin was measured in 389 patients undergoing coronary angiography for a variety of indications at a Veterans Affairs Medical Center. Detailed demographic, clinical, laboratory, and angiographic data were available for patients. LV systolic function was assessed using ventriculography, and patients were grouped into those with normal or mild dysfunction (ejection fraction > or =45%) versus those with moderate to severe systolic dysfunction (ejection fraction <45%). After adjusting for a variety of clinically relevant covariates known to affect LV systolic function, adiponectin was independently associated with LV systolic function in the entire cohort of patients (p = 0.0002) using multivariate linear regression analysis. In addition, using multivariate logistic regression analysis, adiponectin was an independent predictor of the presence of moderate to severe LV dysfunction (odds ratio 1.54, 95% confidence interval 1.21 to 1.97, p = 0.0005). Moreover, baseline adiponectin was also independently associated with LV function in both the myocardial infarction (MI) and non-MI subpopulations of patients (p = 0.0401 and p= 0.0023, respectively). Finally, in the non-MI subpopulation, baseline adiponectin was an independent predictor of moderate to severe LV systolic dysfunction (odds ratio 1.52, 95% confidence interval 1.15 to 2.02, p = 0.0034). In conclusion, baseline plasma adiponectin was an independent predictor of LV systolic dysfunction in a population of patients referred for coronary angiography. PMID- 18394436 TI - Impact of intramural thrombus in coronary arteries on the accuracy of tissue characterization by in vivo intravascular ultrasound radiofrequency data analysis. AB - Virtual Histology (VH) intravascular ultrasound (IVUS) allows differentiation between 4 different tissue phenotypes. However, the current classification tree for analysis cannot differentiate the presence of intramural thrombus. The aim of this study was to evaluate the impact of intramural thrombus for correlative accuracy between in vitro histopathology of coronary atherosclerotic plaque obtained by directional coronary atherectomy and corresponding in vivo tissue characterization obtained by VH IVUS. Coronary IVUS imaging of 30 coronary artery lesions was obtained using a 20-MHz phased-array IVUS catheter with a motorized pull-back system at set 0.5 mm/s. The debulking region of the in vivo histologic image was predicted from comparison between pre- and post-first debulking VH IVUS images. Cross-sectional histologic slices were cut every 0.5 mm starting from the most proximal part of the formalin-fixed debulking tissue. Histologic slices were divided into 2 groups by the presence or absence of pathologic thrombus. A total of 259 in vitro histologic slices were obtained, and pathologic thrombus was detected in 81 slices. Correlation was favorable, with high sensitivity for all plaque components, but specificities for fibrous (thrombus slices vs nonthrombus slices 36% vs 94%) and fibrofatty (9% vs 60%) tissue were lower in thrombus slices. Therefore, predictive accuracies for the 2 plaque components were lower in thrombus slices (fibrous tissue 78% vs 99%, fibrofatty tissue 68% vs 83%, respectively). In conclusion, intramural thrombus was colored as fibrous or fibrofatty by VH IVUS, reducing VH accuracy in these kinds of lesions. PMID- 18394437 TI - Effect of an American Heart Association Get With the Guidelines program-based clinical pathway on referral and enrollment into cardiac rehabilitation after acute myocardial infarction. AB - Cardiac rehabilitation (CR)/secondary prevention programs are an important part of patient care after acute myocardial infarction (AMI). However, only 10% to 15% of eligible patients enroll in such programs. The purpose of this study was to evaluate the effect of an American Heart Association Get With the Guidelines (GWTG)-based clinical pathway on referral and enrollment into CR after AMI. Patients (n = 780) admitted to a single center during an 18-month period with AMI and discharged to home were evaluated retrospectively for referral and enrollment into CR programs. A total of 714 patients (92%) were on the GWTG pathway; 392 (55%) were referred and 135 (19%) were enrolled into CR. Higher referral was associated with pathway use (odds ratio [OR] 2.3, 95% confidence interval [CI] 1.1 to 4.9, p = 0.03), percutaneous coronary intervention (OR 3.1, 95% CI 1.9 to 5.2, p <0.0001), and in-patient physical therapy consultation (OR 13, 95% CI 8.2 to 20.5, p <0.0001). Ethnicity did not affect referral, but was the only variable associated with lower enrollment. Hispanic and black patients had 92% (OR 0.08, 95% CI 0.01 to 0.55, p = 0.02) and 57% (OR 0.43, 95% CI 0.19 to 1.05, p = 0.06) lower odds to enroll compared with white patients, respectively. In conclusion, use of the American Heart Association GWTG pathway showed a significantly higher referral rate to CR after AMI than previously reported in the literature. Nonetheless, most referred patients did not enroll. Strategies to bridge the gap between referral and enrollment in CR should be incorporated into AMI clinical pathways, with special emphasis on increasing enrollment in ethnic minorities. PMID- 18394438 TI - Effect of cytochrome p450 polymorphisms on platelet reactivity after treatment with clopidogrel in acute coronary syndrome. AB - Genetic polymorphisms of cytochrome P450 (CYP) isoforms may promote variability in platelet response to clopidogrel. This study was conducted to analyze, in 603 patients with non-ST elevation acute coronary syndromes, the effect of CYP3A4, CYP3A5, and CYP2C19 gene polymorphisms on clopidogrel response and post-treatment platelet reactivity assessed by adenosine diphosphate (ADP)-induced platelet aggregation, vasodilator-stimulated phosphoprotein phosphorylation index, and ADP induced P-selectin expression. The CYP2C19*2 polymorphism was significantly associated with ADP-induced platelet aggregation, vasodilator-stimulated phosphoprotein phosphorylation index, and ADP-induced P-selectin expression in recessive (p <0.01, p <0.007, and p <0.06, respectively) and codominant (p <0.08, p <0.0001, and p <0.009, respectively) models, but the CYP3A4*1B and CYP3A5*3 polymorphisms were not. The CYP2C19*2 allele carriers exhibited the highest platelet index levels in multivariate analysis (p = 0.03). After covariate adjustment, the CYP2C19*2 allele was more frequent in clopidogrel nonresponders, defined by persistent high post-treatment platelet reactivity (ADP-induced platelet aggregation >70%; p = 0.03). In conclusion, the present data suggest that the CYPC19*2 allele influences post-treatment platelet reactivity and clopidogrel response in patients with non-ST elevation acute coronary syndromes. PMID- 18394439 TI - Anti-cardiolipin antibodies and endothelial function in patients with coronary artery disease. AB - Endothelial dysfunction is considered an important marker in atherosclerosis, having a prognostic value. Antiphospholipid antibodies are considered prothrombotic and have recently been reported to be associated also with atherosclerosis. This study was conducted to investigate a possible association of endothelial dysfunction with various antiphospholipid autoantibodies in healthy subjects and patients with cardiovascular disease. In a single-center, prospective study, 2 groups were included. The study group included patients with cardiovascular diseases (coronary disease and/or cerebrovascular disease) and healthy subjects without apparent heart disease who were referred to the endothelial function laboratory for the assessment of endothelial function. Flow mediated dilatation, which indicates endothelial function, and nitroglycerin mediated vasodilatation, which indicates smooth-muscle function, were measured. The 2 groups were evaluated for autoantibodies, including anticardiolipin (aCL; immunoglobulin G [IgG], immunoglobulin M [IgM], and immunoglobulin A [IgA]), antinuclear antibody, anti-beta2-glycoprotein I (IgG, IgM, and IgA), and oxidized low-density lipoprotein. One hundred seven subjects were included in the study: 45 patients (42%) and 62 healthy controls (58%). Flow-mediated dilatation was significantly lower in patients compared with healthy controls (8.0 +/- 9.5% vs 8.0 +/- 13.5%, p = 0.012). In addition, nitroglycerin-mediated vasodilatation was nonsignificantly lower in patients than in healthy controls (8.0 +/- 13.4% vs 11.0 +/- 16.7%, p = 0.084). The mean levels of anti-beta2-glycoprotein I (IgG, IgM, and IgA), aCL (IgM and IgA), antinuclear antibody, and oxidized low-density lipoprotein were not different between groups. However, the mean level of IgG aCL was significantly higher in patients than in healthy controls. In conclusion, in accordance with previous reports of an association between aCL and atherosclerosis, patients with cardiovascular disease had endothelial dysfunction and elevated levels of aCL. PMID- 18394440 TI - Achievement of therapeutic goals and utilization of evidence-based cardiovascular therapies in coronary heart disease patients with chronic kidney disease. AB - To evaluate whether the presence of chronic kidney disease (CKD) influenced the rate of prescription of evidence-based cardiovascular preventive therapies and attainment of therapeutic goals in patients with stable coronary heart disease, 7,884 patients (mean age 65.4 years; 81.7% men; 22.4% with CKD) attended to in 1,799 primary-care centers and who had had a coronary event requiring hospitalization in the previous 6 months to 10 years were recruited. Glomerular filtration rate (GFR) was estimated using the MDRD Study equation. Results indicated that patients with CKD received more diuretics (47.6% vs 32.8%; p = 0.034), calcium channel blockers (29.3% vs 23.2%, p = 0.027); and blockers of the angiotensin-renin system (76.4% vs 65.3%; p <0.001). The lower prescription rate of antiaggregants, beta blockers, and statins in subjects with CKD did not reach statistical significance in multivariate analysis. A lower percentage of subjects with CKD achieved good control of blood pressure (39.2% vs 65.4%; p <0.001) and glycosylated hemoglobin (43.9% vs 53.4%; p <0.001) relative to patients without CKD. Only 11.8% of patients with CKD had optimum control of all risk factors. Using multivariate analysis, the presence of CKD was inversely related to the degree of risk-factor control, especially in groups with low GFR. In conclusion, patients with stable coronary heart disease and CKD attended to in primary-care centers had poorer control of coronary heart disease risk factors than those with normal GFR despite receiving a similar rate of prescription of evidence-based cardiovascular disease preventive therapies. PMID- 18394441 TI - Relation of bone mineral density to frequency of coronary heart disease. AB - Coronary angiography was performed because of chest pain in 198 patients (146 women, 52 men; mean age 66 years) who had dual-energy x-ray absorptiometry scans of the spine and left hip because of suspected osteoporosis or osteopenia. Of the 198 patients, 53 (27%) had osteoporosis, 79 (40%) had osteopenia, and 66 (33%) had normal bone mineral density (BMD). Obstructive coronary artery disease with >50% narrowing of > or =1 major coronary artery was present in 40 of 53 patients (76%) with osteoporosis, in 54 of 79 patients (68%) with osteopenia, and in 31 of 66 patients (47%) with normal BMD (p <0.005 comparing osteoporosis with normal BMD, p <0.01 comparing osteopenia with normal BMD). In conclusion, in patients who undergo coronary angiography because of chest pain, patients with osteoporosis or osteopenia have a higher prevalence of obstructive coronary artery disease than those with normal BMD. PMID- 18394442 TI - Four-year clinical follow-up of the rapamycin-eluting stent evaluated at Rotterdam Cardiology Hospital registry. AB - Although the safety of drug-eluting stents has been under considerable scrutiny, limited real-world follow-up data extending up to 4 years are available. The randomized clinical trials carefully selected patients and are not reflective of everyday practice. From April to October 2002, 508 consecutive patients treated with sirolimus-eluting stents (SES) were enrolled. The control group consisted of 450 patients treated with bare-metal stents during the preceding 6 months. After 4 years of follow-up, the incidence of composite major adverse clinical events (all-cause death, myocardial infarction, or target vessel revascularization) was found to be significantly lower in the SES group (23.0% vs 28.7%, adjusted hazard ratio 0.66, 95% confidence interval 0.51 to 0.86), as were rates of target vessel revascularization (12.2% vs 17.8%, adjusted hazard ratio 0.57, 95% confidence interval 0.39 to 0.83). There were no differences in all-cause mortality (10.5% for SES vs 10.6% for bare-metal stents, p = 0.9) or in the rates of cardiac death (4.5% vs 6.9%, p = 0.1). Although there was no difference in overall stent thrombosis (2.3% vs 2.2%, p = 1.0), SES had a higher rate of very late stent thrombosis (1.4% vs 0%, p = 0.02), balanced by a lower rate of early stent thrombosis (0.4% vs 1.8%, p = 0.05). In conclusion, after 4 years, SES were found to remain safe and effective compared with bare-metal stents. Nevertheless, the higher rate of very late stent thrombosis remains a concern. Longer term follow up will be required to determine the extent of this problem. PMID- 18394443 TI - Influence of the timing of cardiac catheterization and the amount of contrast media on acute renal failure after cardiac surgery. AB - Postoperative acute renal failure (ARF) is not uncommon after cardiac surgery and after angiography. However, limited information exists regarding the influence of the interval between cardiac catheterization and subsequent cardiac surgery and amount of contrast agent used during this procedure on the occurrence of postoperative ARF. Data for 423 consecutive adult patients who underwent elective cardiac surgery after cardiac catheterization were examined retrospectively. The influence of interval between cardiac catheterization and cardiac surgery on postoperative ARF (defined as postoperative serum creatinine > or =2 times baseline and >2 mg/dl and/or need for renal replacement therapy) was evaluated using multivariable logistic regression. ARF occurred in 24 patients (5.7%). Median time to angiography was 2 days (interquartile range 1 to 4.5), and median dose of contrast used was 1.36 ml/kg (interquartile range 1.12 to 1.69). Surgery on the day of cardiac catheterization was independently associated with increased risk of ARF (adjusted odds ratio 3.1, 95% confidence interval 1.1 to 8.8). This risk of ARF was highest in patients who underwent surgery on the same day as angiography and with a dose of contrast higher than median (14.6%) and lowest when surgery was delayed beyond 1 day of angiography and contrast dose was median or less (2.4%; adjusted odds ratio for same-day surgery and dose higher than median 4.2, 95% confidence interval 1.2 to 14.2). Cardiac surgery performed on the day of cardiac catheterization and higher dose of contrast agent used were both independently associated with increased risk of postoperative ARF. In conclusion, these findings suggest that delaying cardiac surgery beyond 24 hours of exposure to contrast agents (when feasible) and minimizing the use of these agents have significant potential to decrease the incidence of postoperative ARF in patients undergoing elective cardiac surgery. PMID- 18394444 TI - The editor's roundtable: psoriasis, inflammation, and coronary artery disease. PMID- 18394445 TI - Relation of the metabolic syndrome to quantity of coronary atherosclerotic plaque. AB - Although metabolic syndrome (MS) is associated with adverse cardiovascular outcomes, its association with the presence and extent of coronary atherosclerotic plaques is not well described. To assess this relation, multidetector computed tomography-based patterns of coronary plaque were assessed in 77 patients enrolled in the ROMICAT study (age 54 +/- 12 years; 79% Caucasians, and 36% women) and compared between those who did (n = 35; 45%) and did not (n = 42; 55%) have MS. The presence of any, calcified, and noncalcified plaque was significantly higher in patients with than without MS (91%, 74%, and 77% vs 46%, 45%, and 40% segments with plaque, respectively; all p <0.01). The overall number of segments with plaques was also higher in patients with MS (5.8 +/- 3.7 vs 2.1 +/- 3.3; p <0.001). MS was independently associated with both the presence and extent of overall plaques after adjusting for the Framingham risk score (odds ratio 6.7, 95% confidence interval 1.6 to 28.8, p <0.01 for presence, beta coefficient = 3.59 +/- 0.88 [SE], p = 0.009 for extent) and individual risk factors, including age, gender, smoking, body mass index, hypertension, diabetes, hyperlipidemia, and clinical coronary disease (odds ratio 8.4, 95% confidence interval 1.7 to 42.5, p = 0.008 for presence, beta coefficient = 2.35 +/- 0.86 [SE], p = 0.007 for extent). Similarly, MS was independently associated with calcified and noncalcified plaques individually. In conclusion, MS was independently associated with the presence and extent of both calcified and noncalcified coronary atherosclerotic plaques detected using multidetector computed tomography. These data may explain the higher cardiovascular risk in these patients and may lay the foundation for studies to determine whether such information may improve risk stratification. PMID- 18394447 TI - Rhythm versus rate control in the contemporary management of atrial fibrillation in-hospital. AB - Little is presently known regarding whether a rhythm-control or a rate-control strategy is more frequently used in patients hospitalized for atrial fibrillation (AF). This study was conducted to assess patient and physician characteristics associated with each treatment strategy and with the use of anticoagulants. Hospitalizations for primary diagnoses of AF were examined using hospital claims from January 2000 to December 2004. Patients who received antiarrhythmic drugs, ablation, or cardioversion for AF were categorized as receiving rhythm control. Patients managed only with beta blockers, calcium channel blockers, or digoxin were categorized as receiving rate control. Characteristics associated with rhythm compared with rate control and anticoagulant use with CHADS(2) score were determined. The study cohort included 155,731 hospitalizations from 464 hospitals. Of these, 75,397 (48%) were categorized as involving rhythm control and 80,334 (52%) as involving rate control. Care by a noncardiologist (adjusted odds ratio [OR] 0.33, 95% confidence interval [CI] 0.31 to 0.36) and increasing age >65 years (adjusted OR 0.87, 95% CI 0.86 to 0.88) were associated with lower odds of rhythm versus rate control; hypertrophic cardiomyopathy was associated with greater odds (adjusted OR 2.3, 95% CI 1.81 to 2.84) of rhythm control. Warfarin use was greater in the rhythm-control group compared with the rate control group (adjusted OR 1.56, 95% CI 1.52 to 1.60), and warfarin use was greater with a CHADS(2) score > or =2 (unadjusted OR 1.21, 95% CI 1.19 to 1.24). In conclusion, rhythm- and rate-control strategies were used equally in patients hospitalized for AF. Some observations, such as greater use of the rate-control strategy with increasing age, were consistent with recommendations, but others, such as lower use of warfarin in the rate-control group, were not. PMID- 18394446 TI - Inflammatory markers and progression of subclinical atherosclerosis in healthy postmenopausal women (from the Estrogen in the Prevention of Atherosclerosis Trial). AB - The objective of this study was to determine whether high-sensitivity C-reactive protein (hs-CRP) and serum soluble intercellular adhesion molecule-1 (sICAM-1) correlate with progression of subclinical atherosclerosis. Secondarily, the long term effect of oral estradiol on hs-CRP and sICAM-1 were determined. Data were analyzed from 180 healthy postmenopausal women aged 45 to 80 years randomly assigned to either unopposed micronized 17beta-estradiol 1 mg/day or placebo in the Estrogen in the Prevention of Atherosclerosis Trial (EPAT). Carotid artery intima-media thickness (CIMT), hs-CRP, and sICAM-1 were measured at baseline and every 6 months thereafter for 2 years. Unopposed 17beta-estradiol significantly increased hs-CRP (p = 0.01) and decreased sICAM-1 compared with placebo (p = 0.04). Changes in hs-CRP and sICAM-1 did not correlate with changes in carotid artery intima-media thickness. In conclusion, although unopposed 17beta-estradiol significantly altered hs-CRP and sICAM-1, neither marker was associated with progression of subclinical atherosclerosis. PMID- 18394448 TI - Exercise-induced polymorphic ventricular tachycardia in adults without structural heart disease. AB - Patients with catecholaminergic polymorphic ventricular tachycardia present at a young age with exercise-induced ventricular arrhythmias (VAs) and may have a positive family history. We describe 8 patients who presented with exercise induced symptoms as adults, have a negative family history, and responded to beta blocker therapy. The study evaluated exercise treadmill electrocardiographic data from patients referred for exercise-induced VA. Inclusion criteria consisted of development of bidirectional, pleomorphic, or polymorphic ventricular tachycardia with exercise, adult age at first onset, negative family history, and no evidence of structural heart disease. We correlated VA configurations with respect to heart rate before and after beta-blocker therapy. Patients displayed a pattern of increasing ventricular complexity with increasing heart rate. The appropriate beta blocker (n = 7) or calcium channel blocker (n = 1) was defined as the dose that resulted in control of symptoms. Three patients showed suppression of VA with sinus tachycardia at peak heart rate. Six patients had decreased VA defined as absence of higher complexity arrhythmias. With drug therapy, average heart rate associated with premature ventricular complex couplets/triplets increased, whereas duration and complexity of premature ventricular complexes decreased. One patient had an automatic implantable cardiac defibrillator placed but has had no discharges from the device since starting the appropriate beta blocker. In conclusion, these patients appear to respond well to beta-blocker or calcium channel blocker therapy with decreased ectopic complexity and an increased heart rate threshold for inducing VA. PMID- 18394449 TI - Relation of daytime bradyarrhythmias with high risk features of sleep apnea. AB - Bradyarrhythmias (BA) have been reported in patients with sleep apnea (SA), but the incidence of SA in patients with BA remains unclear. A case-control study was conducted to assess the prevalence of high-risk features of SA in patients with documented BA on 24-hour Holter monitoring compared with patients without BA. Controls were age-matched patients selected from those with no evidence of BA on 24-hour Holter monitoring. BA were defined as the presence of pauses of >3 seconds, regardless of the mechanism, and/or heart rate <40 beats/min during presumed waking hours (8 a.m. to 8 p.m.). High-risk features of SA were determined by the Berlin Questionnaire, with positive results defined as having '2 of 3 positive high-risk categories. Body mass index (BMI), hypertension, beta blocker use, and other underlying characteristics were cataloged. Nineteen patients with documented BA and 47 with no BA were identified. The mean ages and BMIs in the active and control groups were not statistically significant. High risk features for SA were present in 57.8% of patients in the BA group compared with 21.3% in the control group (p = 0.003). After controlling for age, BMI, hypertension, and beta-blocker use, patients with BA were 6 times more likely to have high-risk features of SA compared with those without BA (logistic regression odds ratio 6.1, 95% confidence interval 1.5 to 24, p = 0.012). In conclusion, irrespective of BMI, age, and other underlying risk factors, the presence of daytime BA was highly associated with high-risk features of SA. PMID- 18394450 TI - Comparison of clinical features and outcomes of patients hospitalized with heart failure and normal ejection fraction (> or =55%) versus those with mildly reduced (40% to 55%) and moderately to severely reduced (<40%) fractions. AB - Heart failure (HF) with normal ejection fraction (EF) is an increasingly common presentation of acute decompensated HF. Differences between patients with HF and truly normal EF and those with mildly impaired EF have not been described. The Acute Decompensated Heart Failure Registry (ADHERE) contains information on >100,000 HF hospitalizations and may provide insight into this distinction. The ADHERE database was used to investigate differences between patients hospitalized with HF and severely (<25%), moderately (25% to 40%), and mildly (40% to 55%) decreased EF and those with normal EF (> or =55%). The group with normal EF was 69% women with a mean age of 74 years (p <0.0001 vs all other groups). Coronary artery disease was less frequent in the normal EF group, and hypertension played a larger role. Patients with EF > or =55% had increased pulse pressure, suggesting a role for arterial stiffening. Treatment differed by EF. Creatinine increased > or =0.5 mg/dl more often in the group with HF and normal EF than in the group with HF and severely decreased EF. In-hospital mortality and length of stay in the intensive care unit varied inversely with EF; overall length of stay was similar. In conclusion, patients with HF and normal EF are more likely to be women, have a history of high pulse pressure hypertension, less coronary artery disease, and a lower risk of inpatient death but a higher likelihood of deterioration in renal function during hospitalization. These observations may be important considerations in the design of future clinical trials. PMID- 18394451 TI - Usefulness of N-terminal pro-B-type natriuretic peptide increase with exercise for predicting cardiovascular mortality in patients with heart failure. AB - N-terminal pro-B-type natriuretic peptide (NT-pro-BNP) level at rest is related to left ventricular (LV) function and cardiovascular mortality in patients with heart failure (HF). There are limited and controversial data regarding changes in NT-pro-BNP level during exercise in patients with HF. The aim of this study was to investigate the effects of exercise on NT-pro-BNP levels and the relation between increases in NT-pro-BNP and the LV ejection fraction and cardiovascular mortality in patients with HF. Seventy-five patients with HF (New York Heart Association classes I to III) and 20 healthy subjects were enrolled in the study. Echocardiographic examination was performed. The modified Bruce protocol was used for symptom-limited exercise testing. Levels of NT-pro-BNP were measured at rest and after peak exercise. The patients were followed up for 690 to 840 days for cardiovascular mortality. Exercise induced significant increases in NT-pro-BNP in patients and controls. Except for a relative increase in NT-pro-BNP during exercise (relative DeltaNT-pro-BNP), NT-pro-BNP concentrations at rest and during peak exercise and absolute increases in NT-pro-BNP during exercise (absolute DeltaNT-pro-BNP) were significantly higher in patients with HF (p <0.001). Absolute DeltaNT-pro-BNP was positively correlated with NT-pro-BNP at rest (p <0.001). The level of absolute DeltaNT-pro-BNP was the most important parameter in predicting a LV ejection fraction <30% (p <0.001). Absolute DeltaNT-pro-BNP and LV end-systolic volume were found to be independent predictors of mortality (p = 0.012 and p = 0.015, respectively). In conclusion, exercise induced increases in NT-pro-BNP in patients and healthy subjects. Absolute increase in NT pro-BNP is a reliable parameter in predicting a low LV ejection fraction and may help in the identification of patients at high risk for mortality. PMID- 18394452 TI - Effect of mechanical dyssynchrony and cardiac resynchronization therapy on left ventricular rotational mechanics. AB - Alterations in rotational mechanics can bring new aspects to the understanding of left ventricular (LV) dyssynchrony. The aims of this study were to investigate LV rotational mechanics in candidates for cardiac resynchronization therapy (CRT) and to assess the effect of CRT by speckle-tracking echocardiography. Fifty-four patients with heart failure and 33 healthy controls were studied. Thirty-three underwent CRT. Speckle tracking was applied to short- and long-axis views. Radial and longitudinal dyssynchrony were assessed as previously defined. Apical and basal rotations were measured as the average angular displacement about the LV central axis. LV twist and torsion were then calculated. Peak apical and basal rotation, peak LV twist and torsion, apical and basal rotation at aortic valve closure (AVC), and LV twist and torsion at AVC were significantly lower in patients than controls. Apical-basal rotation delay and AVC-to-peak LV twist interval were longer in patients and associated with decreased peak LV twist and LV twist at AVC, respectively. In patients, rotational indexes, particularly LV twist and torsion, were correlated strongly with radial dyssynchrony. LV torsion (cutoff 0.1 degrees /cm) and twist (cutoff 1 degrees ) at AVC had the highest sensitivity (90%) and specificity (77%) to predict CRT responders among all other parameters, including radial and longitudinal dyssynchrony. In conclusion, LV dyssynchrony is associated with discoordinate rotation of the apical and basal regions, which in turn significantly decreases peak LV twist and torsion and LV twist and torsion at AVC. CRT significantly restored the altered rotational mechanics in responders. These parameters have potential for predicting responders to CRT. PMID- 18394454 TI - Management of prosthetic valve infective endocarditis. AB - This study analyzed the profile and outcome of surgically versus medically treated patients with prosthetic valve infective endocarditis (PVE). From 2000 to 2006, 80 patients >16 years of age (median 71) with definite PVE according to modified Duke criteria were included. The medically treated group was separated into deliberately conservative and perforce conservative treatments, the latter group including patients with contraindications to a cardiosurgical intervention. The most frequent causative micro-organisms were staphylococci. Forty-six percent of patients were surgically treated, 34% had deliberately conservative treatment, and 20% had perforce conservative treatment. Six-month mortality was 29%; 27% of surgically treated patients died, 4% deliberately conservatively patients died, and 75% perforce conservatively treated patients died. Septic shock, multiorgan failure, and type of treatment were significantly associated with death in univariable analysis. Multivariable analysis revealed that type of treatment (perforce conservative) and septic shock predicted death in patients with PVE. Survival was most favorable in deliberately conservatively treated patients, including PVE due to Staphylococcus aureus. In conclusion, there remains a role for watchful waiting in patients with PVE without evidence of major complications. Moreover, patients with uncomplicated S. aureus PVE can be treated successfully without cardiac surgery. Conversely, patients with major complicated PVE should preferentially undergo surgery. Predictors of mortality in patients with PVE included septic shock and perforce conservative treatment. PMID- 18394453 TI - Clinical significance of high-density lipoprotein cholesterol in left-sided infective endocarditis. AB - Decreased serum levels of high-density lipoprotein (HDL) cholesterol have been shown to be of prognostic significance in patients with severe infectious diseases. Serum HDL cholesterol levels were therefore investigated as a possible parameter for the prediction of clinical outcomes in patients with left-sided infective endocarditis (IE). Fifty-four patients with IE with available admission serum HDL cholesterol levels were included in the study. A clinical outcome was defined as a complicated course during hospitalization. Forty-two patients had complicated courses during their in-hospital stays. The median serum HDL cholesterol level was significantly lower in patients with IE (n = 54) than healthy controls (n = 26) (26 vs 47 mg/dl, p <0.0001). In the 42 patients with complicated courses, the median serum HDL cholesterol level was lower compared with that in 12 patients with uneventful courses (24 vs 36 mg/dl, p = 0.011). A cut point of serum HDL cholesterol level of 25 mg/dl had sensitivity of 62%, specificity of 75%, and a positive predictive value of 90% for predicting clinical outcomes. In conclusion, serum HDL cholesterol levels measured at admission were markedly reduced in patients with left-sided IE. Furthermore, low serum HDL cholesterol levels predicted complicated clinical courses in these patients. PMID- 18394455 TI - Effects of percutaneous transluminal septal myocardial ablation for obstructive hypertrophic cardiomyopathy on systolic and diastolic left ventricular function assessed by pressure-volume loops. AB - The aim of the present study was to determine the long-term effects of percutaneous transluminal septal myocardial ablation (PTSMA) on systolic and diastolic left ventricular (LV) functions in patients with obstructive hypertrophic cardiomyopathy (HC). Ten consecutive patients with symptomatic HC despite optimal medical treatment were referred for PTSMA at our center. LV systolic and diastolic functions were assessed by online LV pressure-volume loops obtained by conductance catheter at baseline and at 6 months after the procedure. At follow-up, the mean gradients at rest and after extrasystole were significantly decreased compared with baseline (88 +/- 29 to 21 +/- 11 mm Hg and 130 +/- 50 to 35 +/- 22 mm Hg, respectively, p <0.01 for the 2 comparisons). End systolic and end-diastolic pressures significantly decreased (p <0.01), whereas end-systolic and end-diastolic LV volumes significantly increased (p <0.01 for the 2 comparisons). Cardiac output and stroke volume were unchanged, as were ejection fraction (p = 0.25) and maximum dP/dt (p = 0.13). The slope of the end systolic pressure-volume relation was not decreased, indicating a preserved contractility. The relaxation constant time, end-diastolic stiffness, projected volume of the end-diastolic pressure-volume relation at 30 mm Hg, and diastolic stiffness constant showed a significant improvement of active and passive myocardial diastolic properties. In conclusion, PTSMA is an effective method in the treatment of symptomatic patients with HC. At 6-month follow-up, the LV aortic gradient was decreased and active and passive LV diastolic properties were increased. Myocardial contractility was not decreased and general hemodynamics was maintained. PMID- 18394456 TI - Significance of plasma levels of N-terminal Pro-B-type natriuretic peptide on left ventricular remodeling in non-obstructive hypertrophic cardiomyopathy attributable to the Asp175Asn mutation in the alpha-tropomyosin gene. AB - Hypertrophic cardiomyopathy (HC) is an inherited heart disease characterized by left ventricular (LV) remodeling. The present study was conducted to investigate the association of N-terminal pro-B-type natriuretic peptide (NT-pro-BNP) levels with LV remodeling on magnetic resonance imaging and procollagen formation in 17 healthy controls and 24 patients with nonobstructive HC attributable to an identical Asp175Asn (aspartic acid to asparagine at codon 175) mutation in the alpha-tropomyosin gene. None of the patients had history of decompensated heart failure, and all patients had normal LV ejection fraction. Patients with HC had higher NT-pro-BNP levels compared with controls (median 60 pmol/L, range <40 to 359, vs <40 pmol/L; p <0.001), but 9 patients with HC had normal NT-pro-BNP levels (<40 pmol/L). In patients with HC, levels of NT-pro-BNP were correlated significantly with LV end-systolic volume index (r = 0.50, p <0.05), LV mass index (r = 0.47, p <0.05), proportion of hypokinetic segments (r = 0.50, p <0.05), and levels of serum aminoterminal propeptide of type III procollagen (r = 0.52, p <0.01). When patients with HC were divided into 3 groups on the basis of their NT-pro-BNP levels, there were statistically significant linear associations of LV end-systolic volume (test for linearity p = 0.034), LV mass index (p = 0.009), proportion of hypokinetic segments (p = 0.016), and levels of serum aminoterminal propeptide of type III procollagen (p = 0.020) with NT-pro-BNP levels over the 3 groups, suggesting a tight relation between LV remodeling and levels of NT-pro-BNP. In conclusion, in patients with nonobstructive HC attributable to an Asp175Asn mutation in the alpha-tropomyosin gene, elevated NT pro-BNP levels are associated with incipient LV remodeling, suggesting that NT pro-BNP could be used to diagnose insidious unfavorable LV remodeling in HC. PMID- 18394457 TI - Evaluation of mechanical dyssynchrony in children with idiopathic dilated cardiomyopathy and associated clinical outcomes. AB - We studied mechanical dyssynchrony and its association with clinical status in children with idiopathic dilated cardiomyopathy (IDC). The SD of QRS to peak systolic velocity interval by tissue Doppler was measured in 12 left ventricular segments, as a dyssynchrony index (DI), in each child with IDC during a 12-month period. Results were compared with a control cohort. We used the adult-defined DI cutpoint of 32.6 ms to define patients with IDC as "dyssynchronous" or "synchronous" and compared clinical status and outcomes (transplantation listing/death) between these groups. Patients with IDC (n = 23) and controls (n = 14) had similar age, gender, and QRS duration. Patients with IDC had a higher DI than controls (44.8 +/- 23.7 vs 19.9 +/- 8 ms, p <0.0001). A DI >32.6 ms defined mechanical dyssynchrony in 65% of patients with IDC. Dyssynchronous and synchronous patients had similar QRS durations. Age at diagnosis, at dyssynchrony evaluation, and duration of clinical illness were similar in the 2 groups. New York Heart Association score was better in dyssynchronous than in synchronous patients (2 vs 3.1, p <0.05). Number of synchronous and dyssynchronous patients reaching the end point of death or transplantation was similar, although synchronous patients had poorer actuarial survival from the time of diagnosis (hazard ratio 3.25, p = .04). In conclusion, left ventricular mechanical dyssynchrony is prevalent in pediatric IDC. QRS duration alone is inadequate to define dyssynchrony in pediatric IDC, whereas the adult-derived DI of >32.6 ms seems applicable to the pediatric population. In this cohort, the presence of mechanical dyssynchrony was not associated with more severe clinical status or adverse outcomes. PMID- 18394458 TI - Anemia as an independent predictor of perioperative and long-term cardiovascular outcome in patients scheduled for elective vascular surgery. AB - Anemia is common in patients scheduled for vascular surgery and is a risk factor for adverse cardiac outcome. However, it is unclear whether this is an independent risk factor or an expression of underlying co-morbidities. In total, 1,211 patients (77% men, 68 +/- 11 years of age) were enrolled. Anemia was defined as serum hemoglobin levels <13 g/dl for men and <12 g/dl for women and was divided into tertiles to compare mild (men 12.2 to 13.0, women 11.2 to 12.0), moderate (men 11.0 to 12.1, women 10.2 to 11.1), and severe (men 7.2 to 11.0, women 7.5 to 10.1) anemia with nonanemia. Outcome measurements were 30-day and 5 year major adverse cardiac events (MACEs; cardiac death or myocardial infarction). All risk factors were noted. Multivariable logistic and Cox regression analyses were used, adjusting for all cardiac risk factors, including heart failure and renal disease. Data are presented as hazard ratios with 95% confidence intervals. In total, 74 patients (6%) had 30-day MACEs and 199 (17%) had 5-year MACEs. Anemia was present in 399 patients (33%), 133 of whom had mild anemia, 133 had moderate anemia, and 133 had severe anemia. Presence of anemia was associated with renal dysfunction, diabetes, and heart failure. After adjustment for all clinical risk factors, 30-day hazard ratios for a MACE per anemia group were 1.8 for mild (0.8 to 4.1), 2.3 for moderate (1.1 to 5.4), and 4.7 for severe (2.6 to 10.9) anemia, and 5-year hazard ratios for MACE per anemia group were 2.4 for mild (1.5 to 4.2), 3.6 for moderate (2.4 to 5.6), and 6.1 for severe (4.1 to 9.1) anemia. In conclusion, the presence and severity of preoperative anemia in vascular patients are significant predictors of 30-day and 5-year cardiac events, regardless of underlying heart failure or renal disease. PMID- 18394459 TI - Cardiovascular outcomes in male veterans with rheumatoid arthritis. AB - In men with rheumatoid arthritis (RA), the confounding effect of adverse cardiovascular risk profile on the independent association of RA disease activity score (DAS) and major adverse cardiovascular events (MACEs) continues to be debated. The aim was to analyze the association of RA DAS with MACEs in a prospective cohort of men with RA enrolled in the VARA Registry at the Dallas site from January 2003 to October 2006. All subjects met American College of Rheumatology criteria for RA. All events were obtained by reviewing patient clinical data. DAS was categorized as low, 0 to 3.2; moderate, 3.2 to 5.09; and high, > or =5.1. Of 282 men (mean age 66 +/- 11.1 years), 231 had valid DASs (150, low; 60, moderate; and 21, high DAS) and were followed up for 4.4 +/- 2 years. Ninety-two subjects (32.6%; 95% confidence interval 27 to 38) experienced an MACE, a composite end point of death (9 patients; 10%), acute coronary syndrome (38 patients; 42%), coronary revascularization (47 patients; 49%), new onset heart failure (37 patients; 40%), and stroke (15 patients; 16%). DAS was a significant predictor of MACEs (hazard ratio 1.31, 95% confidence interval 1.1 to 1.6, p = 0.01) independent of traditional risk factors. Compared with patients with low or moderate DASs, patients with high DASs had a lower mean event-free period (35 and 30 vs 19 years, respectively; p = 0.03). In conclusion, in a population of male US veterans aged >50 years, (1) patients with RA were at high risk of MACEs, and (2) RA DAS was a significant predictor of MACEs independent of traditional cardiovascular risk factors. PMID- 18394460 TI - Role of radial strain and displacement imaging to quantify wall motion dyssynchrony in patients with left ventricular mechanical dyssynchrony and chronic right ventricular pressure overload. AB - Left ventricular (LV) deformation with ventricular septal shift is one of the most distinctive echocardiographic observations in patients with chronic right ventricular (RV) pressure overload (PO). However, little is known about the effects of RVPO on LV performance and regional synchrony. Accordingly, our objective was to test the hypothesis that chronic RVPO affects regional wall motion, synchronicity, and global LV function using a novel speckle-tracking approach to quantify and characterize regional LV wall motion dyssynchrony. Displacement and strain imaging echocardiographic studies were performed in 20 patients with RVPO from pulmonary arterial hypertension or pulmonic stenosis (mean age 53 +/- 16 years, New York Heart Association class 2.6 +/- 0.7, and peak RV systolic pressure 73 +/- 28 mm Hg) and 20 age-matched normal subjects (mean age 47 +/- 16 years). Segmental signals from 6 segments around the mid-LV short axis were defined as dyssynchronous if their changes were opposite to that of the global LV signal at each time frame, and overall LV dyssynchrony was calculated as the percentage of dyssynchrony in all 6 segments within the specified time interval from onset of QRS to the end of isovolumic relaxation. RVPO was associated with a large degree of regional dyssynchrony with paradoxical ventricular septal motion observed by displacement imaging (21 +/- 6%, p <0.05 vs control group), which was closely associated with LV eccentricity index (r = 0.79, p <0.05) and LV myocardial performance index with linear regression (r = 0.76, p <0.05). In contrast, strain imaging showed uniform segmental radial thickening in the RVPO group, which was similar to the control group, and suggests that there was no intrinsic LV contractile dyssynchrony. In conclusion, LV wall motion dyssynchrony assessed by displacement imaging, not intrinsic contractile dyssynchrony by strain imaging, coexists with LV chamber deformation with ventricular septal shift and is closely associated with impairment of LV performance. PMID- 18394462 TI - [Editorial]. PMID- 18394461 TI - Subclinical cardiac abnormalities in human immunodeficiency virus-infected men receiving antiretroviral therapy. AB - Although cardiotoxic effects of highly active antiretroviral therapy (HAART) are a growing concern, there is a lack of prospective studies of subclinical involvement of the heart in human immunodeficiency virus (HIV)-infected patients. This study evaluated noninvasively cardiac morphologic characteristics and function in HIV-positive (HIV(+)) men receiving HAART for > or =2 years with no clinical evidence of cardiovascular disease. Echocardiography at rest, including tissue Doppler imaging and exercise testing, were performed in 30 HIV(+) men (age 42.1 +/- 4.7 years, duration of HIV infection 10.4 +/- 4.7 years, duration of HAART 5.3 +/- 2.1 years) and 26 age-matched healthy controls. At rest, HIV(+) patients had similar left ventricular (LV) mass indexed to height(2.7) (40.6 +/- 9.5 vs 37.5 +/- 9.3 g/m; p >0.05), but a higher prevalence of LV diastolic dysfunction (abnormal relaxation or pseudonormal filling pattern in 64% of patients vs 12% of controls; p <0.001). LV systolic function indexes were significantly lower (ejection fraction 60.4 +/- 8.7% vs 66.9 +/- 6.9%; p <0.01, and tissue Doppler imaging peak systolic velocity 11.4 +/- 1.6 vs 13.5 +/- 2.2 cm/s; p <0.001). Pulmonary artery pressure was higher in patients compared with controls (32.1 +/- 5.4 vs 26.1 +/- 6.5 mm Hg; p <0.001). Exercise testing showed decreased exercise tolerance in HIV(+) patients, with no case of myocardial ischemia. In conclusion, subclinical cardiac abnormalities are frequently observed in HIV(+) patients on HAART. The usefulness of systematic noninvasive screening in this population should be considered. GECEM study no. 30: National Agency for AIDS Research (ANRS). PMID- 18394464 TI - The Essence of Memory. Preface. PMID- 18394465 TI - Molecular memory traces. AB - To understand the essence of memory, one must examine the working of the brain on many levels. It is important to find the appropriate level to study the particular aspect of memory under investigation. In this review, I will focus on insights gained from examining memory at the molecular level. I will illustrate these insights with specific examples from examining the molecular and cellular mechanisms underlying long-term facilitation in the marine mollusk Aplysia and long-term potentiation, studied mainly in rodents. In particular, I will discuss how molecular memory traces are formed and focus in detail on what role increasing the level of proteins through protein synthesis and gene expression plays in memory formation. I will point out three important constraints from molecular work that should impact on cognitive modeling of the nervous system: (i) the induction of plasticity depends on the 'state' of the synapse; (ii) there are multiple independent molecular traces formed after experience with different half-lives; and (iii) the requirement for the conjunction of synaptic activation and new protein synthesis implies that new conjunctions are required to induce long-term memory formation. PMID- 18394466 TI - PKMzeta, LTP maintenance, and the dynamic molecular biology of memory storage. AB - How memories persist is a fundamental neurobiological question. The most commonly studied physiological model of memory is long-term potentiation (LTP). The molecular mechanisms of LTP can be divided into two phases: induction, triggering the potentiation; and maintenance, sustaining the potentiation over time. Although many molecules participate in induction, very few have been implicated in the mechanism of maintenance. Understanding maintenance, however, is critical for testing the hypothesis that LTP sustains memory storage in the brain. Only a single molecule has been found both necessary and sufficient for maintaining LTP- the brain-specific, atypical PKC isoform, protein kinase Mzeta (PKMzeta). Although full-length PKC isoforms respond to transient second messengers, and are involved in LTP induction, PKMzeta is a second messenger-independent kinase, consisting of the independent catalytic domain of PKCzeta, and is persistently active to sustain LTP maintenance. PKMzeta is produced by a unique PKMzeta mRNA, which is generated by an internal promoter within the PKCzeta gene and transported to the dendrites of neurons. LTP induction increases new PKMzeta synthesis, and the increased level of PKMzeta then enhances synaptic transmission by doubling the number of postsynaptic AMPA receptors (AMPAR) through GluR2 subunit-mediated trafficking of the receptors to the synapse. PKMzeta mediates synaptic potentiation specifically during the late-phase of LTP, as PKMzeta inhibitors can reverse established LTP when applied several hours after tetanization in hippocampal slices or 1 day after tetanization in vivo. These studies set the stage for testing the hypothesis that the mechanism of LTP maintenance sustains memory storage. PKMzeta inhibition in the hippocampus after learning eliminates the retention of spatial memory. Once the PKMzeta inhibitor has been eliminated, the memory is still erased, but new spatial memories can be learned and stored. Similar results are found for conditioned taste aversion when the inhibitor is injected in the insular neocortex. Thus PKMzeta is the first molecule found to be a component of the long-term memory trace. PMID- 18394467 TI - Understanding the importance of mRNA transport in memory. AB - RNA localization is an important mechanism to sort proteins to specific subcellular domains. In neurons, several mRNAs are localized in dendrites and their presence allows autonomous control of local translation in response to stimulation of specific synapses. Active constitutive and activity-induced mechanisms of mRNA transport have been described that represent critical steps in the establishment and maintenance of synaptic plasticity. In recent years, the molecular composition of different transporting units has been reported and the identification of proteins and mRNAs in these RNA granules contributes to our understanding of the key steps that regulate mRNA transport and translation. Although RNA granules are heterogeneous, several proteins are common to different RNA granule populations, suggesting that they play important roles in the formation of the granules and/or their regulation during transport and translation. About 1-4% of the neuron transcriptome is found in RNA granules and the characterization of bound mRNAs reveal that they encode proteins of the cytoskeleton, the translation machinery, vesicle trafficking, and/or proteins involved in synaptic plasticity. Non-coding RNAs and microRNAs are also found in dendrites and likely regulate RNA translation. These mechanisms of mRNA transport and local translation are critical for synaptic plasticity mediated by activity or experience and memory. PMID- 18394468 TI - Cap-dependent translation initiation and memory. AB - It is widely accepted that changes in gene expression contribute to enduring modifications of synaptic strength and are required for long-term memory. This is an exciting, wide-open area of research at this moment, one of those areas where it is clear that important work is underway but where the surface has just been scratched in terms of our understanding. Much attention has been given to the mechanisms of gene transcription; however, the regulation of transcription is only one route of manipulating gene expression. Regulation of mRNA translation is another route, and is the ultimate step in the control of gene expression, enabling cells to regulate protein production without altering mRNA synthesis or transport. One of the main advantages of this mechanism over transcriptional control in the nucleus lies in the fact that it endows local sites with independent decision-making authority, a consideration that is of particular relevance in neurons with complex synapto-dendritic architecture. There are a growing number of groups that are taking on the challenge of identifying the mechanisms responsible for regulating the process of mRNA translation during synaptic plasticity and memory formation. In this chapter we will discuss what has been discovered with regard to the localization and regulation of mRNA translation during specific types of neuronal activity in the mammalian central nervous system. The data are most complete for cap-dependent translation; therefore, particular attention will be paid to the machinery that initiates cap dependent translation and its regulation during synaptic plasticity as well as the behavioral phenotypes consequent to its dysregulation. PMID- 18394469 TI - Translational control of gene expression: a molecular switch for memory storage. AB - A critical requirement for the conversion of the labile short-term memory (STM) into the consolidated long-term memory (LTM) is new gene expression (new mRNAs and protein synthesis). The first clues to the molecular mechanisms of the switch from short-term to LTM emerged from studies on protein synthesis in different species. Initially, it was shown that LTM can be distinguished from STM by its susceptibility to protein synthesis inhibitors. Later, it was found that long lasting synaptic changes, which are believed to be a key cellular mechanism by which information is stored, are also dependent on new protein synthesis. Although the role of protein synthesis in memory was reported more than 40 years ago, recent molecular, genetic, and biochemical studies have provided fresh insights into the molecular mechanisms underlying these processes. In this chapter, we provide an overview of the role of translational control by the eIF2alpha signaling pathway in long-term synaptic plasticity and memory consolidation. PMID- 18394471 TI - 'Synaptic tagging' and 'cross-tagging' and related associative reinforcement processes of functional plasticity as the cellular basis for memory formation. AB - We focus on new properties of cellular and network processes of memory formation involving 'synaptic tagging' and 'cross-tagging' during long-term potentiation (LTP) and long-term depression (LTD) as well as associative heterosynaptic interactions, the latter of which are characterized by a time-window of about 1h. About 20 years ago we showed for the first time that the maintenance of LTP, like memory storage, depends on intact protein synthesis and thus consists of at least two temporal phases. Later, similar properties for LTD were shown by our own and other laboratories. Here we describe the requirements for the induction of the transient early-LTP/LTD and of the protein synthesis-dependent late-LTP/LTD. Late LTP/LTD depend on the associative activation of heterosynaptic inputs, i.e. the synergistic activation of glutamatergic and modulatory reinforcing inputs within specific, effective time-windows during their induction. The induction of late LTP/LTD is characterized by novel, late-associative properties such as 'synaptic tagging', 'cross-tagging' and 'late-associative reinforcement'. All of these phenomena require the associative setting of synaptic tags as well as the availability of plasticity-related proteins (PRPs) and they are restricted to functional dendritic compartments, in general. 'Synaptic tagging' guarantees input specificity, 'cross-tagging' determines the interaction between LTP and LTD in a neuron, and thus both are required for the specific processing of afferent signals for the establishment of late-LTP/LTD. 'Late-associative reinforcement' describes a process where early-LTP/LTD by the co-activation of modulatory inputs can be transformed into late-LTP/LTD in activated synapses where a tag is set. Recent experiments in the freely moving rat revealed a number of modulatory brain structures involved in the transformation of early-plasticity events into long lasting ones. Further to this, we have characterized time-windows and activation patterns to be effective in the reinforcement process. Studies using a combined electrophysiological and behavioural approach revealed the physiological relevance of these reinforcement processes, which is also supported by fMRI studies in humans, which led to the hypothesis outlined here on cellular and system memory-formation by late-associative heterosynaptic interactions at the cellular level during functional plasticity events. PMID- 18394472 TI - Synaptic plasticity in learning and memory: stress effects in the hippocampus. AB - Synaptic plasticity has often been argued to play an important role in learning and memory. The discovery of long-term potentiation (LTP) and long-term depression (LTD), the two most widely cited cellular models of synaptic plasticity, significantly spurred research in this field. Although correlative evidence suggesting a role for synaptic changes such as those seen in LTP and LTD in learning and memory has been gained in a number of studies, definitive demonstrations of a specific role for either LTP or LTD in learning and memory are lacking. In this review, we discuss a number of recent advancements in the understanding of the mechanisms that mediate LTP and LTD in the rodent hippocampus and focus on the use of subunit-specific N-methyl-d-aspartate receptor antagonists and interference peptides as potential tools to study the role of synaptic plasticity in learning and memory. By using the modulation of synaptic plasticity and hippocampal-dependent learning and memory by acute stress as an example, we review a large body of convincing evidence indicating that alterations in synaptic plasticity underlie the changes in learning and memory produced by acute stress. PMID- 18394470 TI - Regulation of hippocampus-dependent memory by cyclic AMP-dependent protein kinase. AB - The hippocampus is crucial for the consolidation of new declarative long-term memories. Genetic and behavioral experimentation have revealed that several protein kinases are critical for the formation of hippocampus-dependent long-term memories. Cyclic-AMP dependent protein kinase (PKA) is a serine-threonine kinase that has been strongly implicated in the expression of specific forms of hippocampus-dependent memory. We review evidence that PKA is required for hippocampus-dependent memory in mammals, and we highlight some of the proteins that have been implicated as targets of PKA. Future directions and open questions regarding the role of PKA in memory storage are also described. PMID- 18394473 TI - The role of the GluR-A (GluR1) AMPA receptor subunit in learning and memory. AB - It is widely believed that synaptic plasticity may provide the neural mechanism that underlies certain kinds of learning and memory in the mammalian brain. The expression of long-term potentiation (LTP) in the hippocampus, an experimental model of synaptic plasticity, requires the GluR-A subunit of the AMPA subtype of glutamate receptor. Genetically modified mice lacking the GluR-A subunit show normal acquisition of the standard, fixed-location, hidden-platform watermaze task, a spatial reference memory task that requires the hippocampus. In contrast, these mice are dramatically impaired on hippocampus-dependent, spatial working memory tasks, in which the spatial response of the animal is dependent on information in short-term memory. Taken together, these results argue for two distinct and independent spatial information processing mechanisms: (i) a GluR-A independent associative learning mechanism through which a particular spatial response is gradually or incrementally strengthened, and which presumably underlies the acquisition of the classic watermaze paradigm and (ii) a GluR-A dependent, non-associative, short-term memory trace which determines performance on spatial working memory tasks. These results are discussed in terms of Wagner's SOP model (1981). PMID- 18394474 TI - Synaptic remodeling, synaptic growth and the storage of long-term memory in Aplysia. AB - Synaptic remodeling and synaptic growth accompany various forms of long-term memory. Storage of the long-term memory for sensitization of the gill-withdrawal reflex in Aplysia has been extensively studied in this respect and is associated with the growth of new synapses by the sensory neurons onto their postsynaptic target neurons. Recent time-lapse imaging studies of living sensory-to-motor neuron synapses in culture have monitored both functional and structural changes simultaneously so as to follow remodeling and growth at the same specific synaptic connections continuously over time and to examine the functional contribution of these learning-related structural changes to the different time dependent phases of memory storage. Insights provided by these studies suggest the synaptic differentiation and growth induced by learning in the mature nervous system are highly dynamic and often rapid processes that can recruit both molecules and mechanisms used for de novo synapse formation during development. PMID- 18394475 TI - Spine dynamics and synapse remodeling during LTP and memory processes. AB - While changes in the efficacy of synaptic transmission are believed to represent the physiological bases of learning mechanisms, other recent studies have started to highlight the possibility that a structural reorganization of synaptic networks could also be involved. Morphological changes of the shape or size of dendritic spines or of the organization of postsynaptic densities have been described in several studies, as well as the growth and formation following stimulation of new protrusions. Confocal in vivo imaging experiments have further revealed that dendritic spines undergo a continuous turnover and replacement process that may vary as a function of development, but can be markedly enhanced by sensory activation or following brain damage. The implications of these new aspects of plasticity for learning and memory mechanisms are discussed. PMID- 18394476 TI - The age of plasticity: developmental regulation of synaptic plasticity in neocortical microcircuits. AB - Proper wiring of neural circuits during development depends on both molecular cues that guide connectivity and activity-dependent mechanisms that use patterned activation to adjust the strength and number of synaptic connections. In this chapter, we discuss some of the plasticity mechanisms underlying the experience dependent rewiring of visual cortical microcircuits focusing on layer 4 of rat primary visual cortex. The microcircuit in layer 4 has the ability to regulate its excitability by shifting the balance between excitatory and inhibitory synaptic transmission in an experience-dependent manner. Early in postnatal development (shortly after eye opening), visual deprivation activates several forms of homeostatic plasticity that cooperate to adjust layer 4 excitability to compensate for reduced sensory drive. In contrast, during the classical sensitive period for rodent visual system plasticity, this homeostatic response is replaced by mechanisms that reduce the responsiveness of deprived cortex. We discuss this developmentally regulated switch in plasticity within layer 4 and how this might depend on the maturation of excitatory and inhibitory monosynaptic connections. Based on our published data, we propose inhibitory plasticity as an important player in circuit refinement that can contribute both to the compensatory forms of circuit plasticity in the early stages of development and to the pathological loss of function induced by visual deprivation during the critical period. PMID- 18394477 TI - Differential mechanisms of transmission and plasticity at mossy fiber synapses. AB - The last few decades have seen the hippocampal formation at front and center in the field of synaptic transmission. However, much of what we know about hippocampal short- and long-term plasticity has been obtained from research at one particular synapse; the Schaffer collateral input onto principal cells of the CA1 subfield. A number of recent studies, however, have demonstrated that there is much to be learned about target-specific mechanisms of synaptic transmission by study of the lesser known synapse made between the granule cells of the dentate gyrus; the so-called mossy fiber synapse, and its targets both within the hilar region and the CA3 hippocampus proper. Indeed investigation of this synapse has provided an embarrassment of riches concerning mechanisms of transmission associated with feedforward excitatory and inhibitory control of the CA3 hippocampus. Importantly, work from a number of labs has revealed that mossy fiber synapses possess unique properties at both the level of their anatomy and physiology, and serve as an outstanding example of a synapse designed for target specific compartmentalization of synaptic transmission. The purpose of the present review is to highlight several aspects of this synapse as they pertain to a novel mechanism of bidirectional control of synaptic plasticity at mossy fiber synapses made onto hippocampal stratum lucidum interneurons. It is not my intention to pour over all that is known regarding the mossy fiber synapse since many have explored this topic exhaustively in the past and interested readers are directed to other fine reviews (Henze et al., 2000; Urban et al., 2001; Lawrence and McBain, 2003; Bischofberger et al., 2006; Nicoll and Schmitz, 2005). PMID- 18394478 TI - Long-term synaptic plasticity in hippocampal feedback inhibitory networks. AB - Recent studies clearly indicate that long-term synaptic plasticity in hippocampal networks not only takes place at excitatory synapses of hippocampal granule and pyramidal cells, but also at excitatory synapses onto inhibitory interneurons. Various forms of long-term potentiation (LTP) and depression (LTD) have now been reported at glutamatergic synapses of interneurons in dentate gyrus (DG), CA3, and CA1 regions of the hippocampus. Importantly, the presence and type of these changes in synaptic efficacy appear to depend on the interneuron subtype, including its specific role within the hippocampal network. The data reviewed here suggest the existence of cell-type specific rules for synaptic plasticity in hippocampal feed-forward and feedback inhibitory networks. This specialized tuning of inhibition is likely important for global hippocampal function. PMID- 18394480 TI - Animal models and behaviour: their importance for the study of memory. AB - In our overview, we will attempt to justify the use of animal models and suggest that it is the only way to make the successive transitions between changes occurring at the molecular and cellular levels and changes at the level of behaviour in the intact organism. We will also stress the importance of criteria that have to be fulfilled in order to unravel the cellular mechanisms of memory: detectability, mimicry, anterograde alteration and retrograde alteration. We will also propose that a large number of animal models should be used to explore the great variety of potential mechanisms that may exist to explain behaviours and their modifications and in particular memory. Finally using the experimental model of Aplysia as example we will insist that to explain the total reflex in an intact animal, all the neurons - sensory neurons and different layers of excitatory and inhibitory interneurons - have to be investigated. PMID- 18394479 TI - Persistent neural activity in the prefrontal cortex: a mechanism by which BDNF regulates working memory? AB - Working memory is the ability to maintain representations of task-relevant information for short periods of time to guide subsequent actions or make decisions. Neurons of the prefrontal cortex exhibit persistent firing during the delay period of working memory tasks. Despite extensive studies, the mechanisms underlying this persistent neural activity remain largely obscure. The neurotransmitter systems of dopamine, NMDA, and GABA have been implicated, but further investigations are necessary to establish their precise roles and relationships. Recent research has suggested a new component: brain-derived neurotrophic factor (BDNF) and its high-affinity receptor, TrkB. We review the research on persistent activity and suggest that BDNF/TrkB signaling in a distinct class of interneurons plays an important role in organizing persistent neural activity at the single-neuron and network levels. PMID- 18394483 TI - Associative learning signals in the brain. AB - Associative memory is defined as memory for the relationship between two initially unrelated items, like a name and an unfamiliar face. Associative memory is not only one of the most common forms of memory used in everyday situations, but is highly dependent on the structures of the medial temporal lobe (MTL). The goal of this chapter is to review the patterns of neural activity shown to underlie the formation of new associative memories in the MTL, as well as to examine how other extra-MTL areas participate in the learning process. Other areas implicated in various aspects of associative learning include the motor related areas of the frontal lobe, prefrontal cortex, and striatum. The question of how the MTL and the other cortical and subcortical structures may interact during associative learning will be discussed. PMID- 18394481 TI - New tricks for an old slug: the critical role of postsynaptic mechanisms in learning and memory in Aplysia. AB - The marine snail Aplysia has served for more than four decades as an important model system for neurobiological analyses of learning and memory. Until recently, it has been believed that learning and memory in Aplysia were due predominately, if not exclusively, to presynaptic mechanisms. For example, two nonassociative forms of learning exhibited by Aplysia, sensitization and dishabituation of its defensive withdrawal reflex, have been previously ascribed to presynaptic facilitation of the connections between sensory and motor neurons that mediate the reflex. Recent evidence, however, indicates that postsynaptic mechanisms play a far more important role in learning and memory in Aplysia than formerly appreciated. In particular, dishabituation and sensitization depend on a rise in intracellular Ca(2+) in the postsynaptic motor neuron, postsynaptic exocytosis, and modulation of the functional expression of postsynaptic AMPA-type glutamate receptors. In addition, the expression of the persistent presynaptic changes that occur during intermediate- and long-term dishabituation and sensitization appears to require retrograde signals that are triggered by elevated postsynaptic Ca(2+). The model for learning-related synaptic plasticity proposed here for Aplysia is similar to current mammalian models. This similarity suggests that the cellular mechanisms of learning and memory have been highly conserved during evolution. PMID- 18394482 TI - Olfactory memory traces in Drosophila. AB - In Drosophila, the fruit fly, coincident exposure to an odor and an aversive electric shock can produce robust behavioral memory. This behavioral memory is thought to be regulated by cellular memory traces within the central nervous system of the fly. These molecular, physiological, or structural changes in neurons, induced by pairing odor and shock, regulate behavior by altering the neurons' response to the learned environment. Recently, novel in vivo functional imaging techniques have allowed researchers to observe cellular memory traces in intact animals. These investigations have revealed interesting temporal and spatial dynamics of cellular memory traces. First, a short-term cellular memory trace was discovered that exists in the antennal lobe, an early site of olfactory processing. This trace represents the recruitment of new synaptic activity into the odor representation and forms for only a short period of time just after training. Second, an intermediate-term cellular memory trace was found in the dorsal paired medial neuron, a neuron thought to play a role in stabilizing olfactory memories. Finally, a long-term protein synthesis-dependent cellular memory trace was discovered in the mushroom bodies, a structure long implicated in olfactory learning and memory. Therefore, it appears that aversive olfactory associations are encoded by multiple cellular memory traces that occur in different regions of the brain with different temporal domains. PMID- 18394484 TI - What are the differences between long-term, short-term, and working memory? AB - In the recent literature there has been considerable confusion about the three types of memory: long-term, short-term, and working memory. This chapter strives to reduce that confusion and makes up-to-date assessments of these types of memory. Long- and short-term memory could differ in two fundamental ways, with only short-term memory demonstrating (1) temporal decay and (2) chunk capacity limits. Both properties of short-term memory are still controversial but the current literature is rather encouraging regarding the existence of both decay and capacity limits. Working memory has been conceived and defined in three different, slightly discrepant ways: as short-term memory applied to cognitive tasks, as a multi-component system that holds and manipulates information in short-term memory, and as the use of attention to manage short-term memory. Regardless of the definition, there are some measures of memory in the short term that seem routine and do not correlate well with cognitive aptitudes and other measures (those usually identified with the term "working memory") that seem more attention demanding and do correlate well with these aptitudes. The evidence is evaluated and placed within a theoretical framework depicted in Fig. 1. PMID- 18394485 TI - Encoding-retrieval overlap in human episodic memory: a functional neuroimaging perspective. AB - The principle of transfer-appropriate processing and the cortical reinstatement hypothesis are two influential theoretical frameworks, articulated at the psychological and neurobiological levels of explanation, respectively, that each propose that the processes supporting the encoding and retrieval of episodic information are strongly interdependent. Here, we integrate these two frameworks into a single model that generates predictions that can be tested using functional neuroimaging methods in healthy humans, and then go on to describe findings that are in accord with these predictions. Consistent with the transfer appropriate processing and cortical reinstatement frameworks, the neural correlates of successful encoding vary according to how retrieval is cued, and the neural correlates of retrieval are modulated by how items are encoded. Thus, encoding and retrieval should not be viewed as separate stages of memory that can be investigated in isolation from one another. PMID- 18394486 TI - Cognitive aging and increased distractibility: costs and potential benefits. AB - Older adults show a characteristic pattern of impaired and spared functioning relative to younger adults. Elsewhere we have argued that many age-related changes in cognitive function are rooted in an impaired ability to inhibit irrelevant information and inappropriate responses. In this chapter we review evidence that as a direct result of impaired inhibitory processes, older adults tend to be highly susceptible to distraction. We suggest that because the distinction between relevant and irrelevant is seldom either clear or static, distractibility can manifest as either a cost or a benefit depending on the situation. We review evidence that in situations in which it interferes with the current task, distraction is disproportionately detrimental to older adults compared to university aged adults, but that when previously distracting information becomes relevant, older adults show a benefit whereas younger adults do not. PMID- 18394487 TI - Characterizing the memory changes in persons with mild cognitive impairment. AB - Persons with mild cognitive impairment (MCI) do not meet criteria for Alzheimer's disease (AD) but are at high risk for developing the disease. Presence of a memory deficit is a key component in the characterization of MCI. This chapter presents empirical studies that attempt to describe and understand the nature of the memory deficit in MCI with a focus on episodic memory and working memory. Cross-sectional studies report prominent deficits of episodic memory characterized by impaired encoding of the contextual information that makes up complex events. This results in reduced free and cued recall, impaired recognition, and impaired associative learning. Although semantic encoding is found to be impaired in conditions that rely on explicit and intentional retrieval, preserved semantic processing is found with automatic conditions of testing. Studies indicate the presence of a partial deficit of working memory with the ability to divide attention being most severely impaired. However, there appears to be heterogeneity as to the extent of the working memory impairment. The presence of vascular anomalies on MRI, as well as being in a more advanced stage in the continuum from MCI to AD, are associated with more severe and more pervasive working memory deficits. Finally, longitudinal studies indicate that the combination of episodic and working memory deficits represents a strong predictor of progression from MCI to AD. PMID- 18394488 TI - Aging, metamemory regulation and executive functioning. AB - In this chapter we deal with metamemory regulation processes and concentrate mainly on how they are related to learning in episodic memory. In recent years an increasing amount of the literature has emphasized conceptual similarities between metamemory regulation and executive-frontal functioning. Different data have also highlighted that age-related cognitive differences might, in many cases, be explained by the decline of executive-frontal functioning that accompanies aging. Thus, in the present chapter we evaluate the relationship of aging and metamemory regulation among the cognitive decline frontal hypothesis of aging. We focus specifically on two measures of metamemory regulation allowing evaluating monitoring and control processes: feeling-of-knowing (FOK) and capacity to adjust study strategies to task demand, respectively. After having presented evidence supporting the executive-frontal hypothesis of FOK, we present a series of experiments addressing the questions of age-related differences in metamemory monitoring and control, and of possible mediation of this age effect by the age-related decline in executive-frontal functioning. The findings support the ideas that the monitoring process of episodic memory FOK and the control process of adjusting study time to task difficulty are impaired in older adults. Moreover, these declines can be explained by the decline of executive-frontal functioning associated to aging. Finally, types of mechanisms pertaining to FOK monitoring and to adjustment control process on which executive-frontal functioning and aging may have an impact are discussed. PMID- 18394489 TI - Cognitive neuroscience studies of semantic memory in Alzheimer's disease. AB - Semantic memory is the component of long-term memory that stores our concepts about the world. The disruption of semantic memory as a result of brain damage may have profound negative consequences on an individual's ability to name objects and process concepts. This can be disrupted as a result of many forms of brain damage, particularly Alzheimer's disease (AD). The current paper reviews research demonstrating that semantics deteriorates early in AD, particularly on effortful semantic tasks. There is a "category effect", meaning that AD preferentially affects concepts dealing with living things and abstract concepts compared to non-living objects and verbs/actions. While this pattern of deterioration, specific for AD, may reflect a breakdown within a distributed semantic system (where living things are distinguished by a high rate of inter correlations between concepts or by a particular mode of being learned), it is equally possible that there is a regional distribution of semantic knowledge, with living things preferentially involving left temporal regions which become damaged early on in AD. Evidence from patients with strokes and semantic dementia, as well as activation studies in normal individuals, implicates the left posterior temporal region in semantic processing for pictures, abstract words, and concrete words. AD individuals, who are impaired in a variety of semantic tasks, show functional deficits in this area, and fail to activate it normally. PMID- 18394490 TI - The effects of surgery and anesthesia on memory and cognition. AB - This chapter describes current findings from the research into postoperative cognitive dysfunction (POCD) following cardiac and non-cardiac surgery in older adults. The evidence suggests that a significant proportion of patients show POCD in the early weeks following surgery and anesthesia. Specific domains of cognition are affected, especially memory. Much less evidence supports the presence of POCD several months or years after surgery, suggesting that POCD may be transient. However, several methodological issues make it difficult to compare findings across studies. Increasing age is among the most consistently reported patient-related risk factor. Other factors more directly related to the surgery and anesthesia are likely to contribute to the pathogenesis of POCD, including inflammatory processes triggered by the surgical procedure. Animal studies have provided valuable findings otherwise not possible in human studies; these include a correlation between the inflammatory response in the hippocampus and the development of POCD in rodents. PMID- 18394491 TI - Cytochemical and molecular characteristics of the process of cornification during feather morphogenesis. AB - Feathers are the most complex epidermal derivatives among vertebrates. The present review deals with the origin of feathers from archosaurian reptiles, the cellular and molecular aspects of feather morphogenesis, and focus on the synthesis of keratins and associated proteins. Feathers consist of different proteins among which exists a specialized group of small proteins called beta keratins. Genes encoding these proteins in the chick genome are distributed in different chromosomes, and most genes encode for feather keratins. The latter are here recognized as proteins associated with the keratins of intermediate filaments, and functionally correspond to keratin-associated proteins of hairs, nails and horns in mammals. These small proteins possess unique properties, including resistance and scarce elasticity, and were inherited and modified in feathers from ancestral proteins present in the scales of archosaurian progenitors of birds. The proteins share a common structural motif, the core box, which was present in the proteins of the reptilian ancestors of birds. The core box allows the formation of filaments with a different molecular mechanism of polymerization from that of alpha-keratins. Feathers evolved after the establishment of a special morphogenetic mechanism gave rise to barb ridges. During development, the epidermal layers of feathers fold to produce barb ridges that produce the ramified structure of feathers. Among barb ridge cells, those of barb and barbules initially accumulate small amounts of alpha-keratins that are rapidly replaced by a small protein indicated as "feather keratin". This 10 kDa protein becomes the predominant form of corneous material of feathers. The main characteristics of feather keratins, their gene organization and biosynthesis are similar to those of their reptilian ancestors. Feather keratins allow elongation of feather cells among supportive cells that later degenerate and leave the ramified microstructure of barbs. In downfeathers, barbs are initially independent and form plumulaceous feathers that rest inside a follicle. Stem cells remain in the follicle and are responsible for the regeneration of pennaceous feathers. New barb ridges are produced and they merge to produce a rachis and a flat vane. The modulation of the growth pattern of barb ridges and their fusion into a rachis give rise to a broad variety of feather types, including asymmetric feathers for flight. Feather morphogenesis suggests possible stages for feather evolution and diversification from hair-like outgrowths of the skin found in fossils of pro-avian archosaurians. PMID- 18394492 TI - Evidence-based practice in 5 simple steps. AB - This editorial provides a brief review of Sackett's 5 steps of evidence-based practice. PMID- 18394493 TI - Predictors for immediate and global responses to chiropractic manipulation of the cervical spine. AB - OBJECTIVE: Patients with nonspecific musculoskeletal disorders may vary in their response to treatment. This study set out to identify the predictors for either improvement or worsening in symptoms for which cervical spine manipulation is indicated. METHOD: A large prospective study recorded details on patients, their presenting symptoms, and type of treatment. At the end of the consultation, any immediate improvement or worsening in presenting symptoms was noted. At the follow-up visit, information was collected on the patients' self-reported improvement. RESULTS: Data were collected from 28,807 treatment consultations (in 19,722 patients) and 13,873 follow-up treatments. The presenting symptoms of "neck pain," "shoulder, arm pain," "reduced neck, shoulder, arm movement, stiffness," "headache," "upper, mid back pain," and "none or one presenting symptom" emerged in the final model as significant predictors for an immediate improvement. The presence of any 4 of these predictors raised the probability for an immediate improvement in presenting symptoms after treatment from 70% to approximately 95%. With regard to immediate worsening, "neck pain," "shoulder, arm pain, "headache," "numbness, tingling upper limbs," "upper, mid back pain," and "fainting, dizziness, light-headedness" emerged as predictors; and the presence of any 4 of these raised the probability for immediate worsening from 4.4% to approximately 12%. For global improvement, only 2 predictors were identified; but these did not enhance the postprediction probability. CONCLUSIONS: This study is the first attempt to identify variables that can predict immediate outcomes in terms of improvement and worsening of presenting symptoms, and global improvement, after cervical spine manipulation. The predictor variables were strongest for immediate improvement. PMID- 18394494 TI - Postural control in people with osteoarthritis of the cervical spine. AB - OBJECTIVE: This study examines whether there is a relationship between clinical correlates of osteoarthritic changes of the cervical spine and changes in postural stability. METHODS: This was a control group study with repeated measures. Twenty-three patients were recruited from the chiropractic University Clinic to participate in this study. The presence and severity of osteoarthritic changes of the cervical spine were determined radiologically. Balance control was evaluated by testing subjects' postural stability on a force platform with and without vision. A general clinical assessment of the neuromusculoskeletal system was performed to screen for any physical condition that could affect postural stability. Participants' characteristics were compared between each group using a 1-way analysis of variance for independent samples, and postural stability variables were submitted to a 2-way repeated measures analysis of variance. RESULTS: Subjects with signs of osteoarthritis of the cervical spine showed an increased range of sway, a faster sway speed, and a greater excursion than control subjects. They also showed a larger degree of lower limb neuropathy than control subjects. CONCLUSION: The postural instability shown by the osteoarthritic group may be due to the effects of the lower limb peripheral neuropathy alone or due to a combination of both cervical degenerative changes and peripheral changes. Further research is needed to clearly isolate the effects of the degeneration of the cervical spine on postural control. PMID- 18394495 TI - Comparison of 3 physical therapy modalities for acute pain in lumbar disc herniation measured by clinical evaluation and magnetic resonance imaging. AB - OBJECTIVE: This study measures and compares the outcome of traction, ultrasound, and low-power laser (LPL) therapies by using magnetic resonance imaging and clinical parameters in patients presenting with acute leg pain and low back pain caused by lumbar disc herniation (LDH). METHODS: A total of 60 patients were enrolled in this study and randomly assigned into 1 of 3 groups equally according to the therapies applied, either with traction, ultrasound, or LPL. Treatment consisted of 15 sessions over a period of 3 weeks. Magnetic resonance imaging examinations were done before and immediately after the treatment. Physical examination of the lumbar spine, severity of pain, functional disability by Roland Disability Questionnaire, and Modified Oswestry Disability Questionnaire were assessed at baseline, immediately after, and at 1 and 3 months after treatment. RESULTS: There were significant reductions in pain and disability scores between baseline and follow-up periods, but there was not a significant difference between the 3 treatment groups at any of the 4 interview times. There were significant reductions of size of the herniated mass on magnetic resonance imaging after treatment, but no differences between groups. CONCLUSIONS: This study showed that traction, ultrasound, and LPL therapies were all effective in the treatment of this group of patients with acute LDH. These results suggest that conservative measures such as traction, laser, and ultrasound treatments might have an important role in the treatment of acute LDH. PMID- 18394496 TI - The refractory period of the audible "crack" after lumbar manipulation: a preliminary study. AB - OBJECTIVE: This study evaluates if side posture lumbar manipulation is associated with a refractory period of the audible "crack" and if so, to quantify this refractory period across subjects. METHODS: Three subjects were exposed to multiple "baseline" side posture manipulations until no further audible cracks were recorded. "Test-refractory period" manipulations were administered after a set time (ie, potential refractory period) at which point the number of audible cracks was recorded. The refractory period was declared when a minimum of 50% of the baseline audible "cracks" had recovered during the test manipulations. The study design included 2 clinicians who performed side posture lumbar manipulation on asymptomatic subjects ranging from 38 to 49 years of age. RESULTS: The refractory period was 40 minutes for subject A, 70 minutes for subject B, and 95 minutes for subject C. The average refractory period across subjects was 68.33 minutes. The audible "crack" recovery was maintained for the remaining test days once the refractory period had been met. CONCLUSIONS: The audible "crack" heard during side posture lumbar manipulation is believed to originate from the zygapophyseal joints. This is supported by the presence of a refractory period and by the number of audible "cracks" found per manipulation. PMID- 18394497 TI - Induced static asymmetry of the pelvis is associated with functional asymmetry of the lumbo-pelvo-hip complex. AB - OBJECTIVE: This study evaluates the hypothesis that triggering and eliminating induced static pelvic asymmetry (SPA) may be followed by immediate change in functional asymmetry of the lumbo-pelvo-hip complex. METHODS: Repeated measures experimental design with 2 levels of independent variable, that is, induced SPA triggered and induced SPA eliminated, was implemented. Three series of measurements were performed, that is, baseline, after triggering SPA, and after eliminating SPA. A group of 84 subjects with no initial symptoms of SPA was studied. Different forms of mechanical stimulation were applied aiming to induce SPA, and the 2 manual stretching-manipulating techniques were performed aiming to eliminate it. A hand inclinometer was used to measure SPA in standing posture. Selected ranges of motion of the hip joints and lumbar spine were used to depict functional asymmetry of the lumbo-pelvo-hip complex. The functional asymmetry indices for individual movements were calculated. Repeated measures design of analysis of variance, dependent data Student t test, and linear Pearson's correlation test were used. RESULTS: Assessment of the SPA showed its significant increase between baseline and series 2 measurements, with a subsequent significant decrease between series 2 and series 3 measurements. Values of the functional asymmetry indices changed accordingly, that is, they increased significantly between series 1 and series 2 and had returned to their initial level in series 3 measurements. CONCLUSIONS: Induced SPA shows considerable association with functional asymmetry of the lumbo-pelvo-hip complex. PMID- 18394498 TI - Principal components analysis of the atlas vertebra. AB - OBJECTIVE: This project explored morphological asymmetry of the atlas by comparing multiple structural variables bilaterally using multivariate statistics. METHODS: Two hundred thirty-seven atlases were obtained from the Hamann-Todd Osteological Collection (Cleveland Museum of Natural History, Cleveland, Ohio). The following dimensions were bilaterally measured: anteroposterior length of the superior articular facet, width of the superior articular facet, length of the transverse process, width of the vertebral canal, anteroposterior diameter of the inferior facet, height of the lateral mass, transverse diameter of the inferior articular facet, width of the inferior lateral mass, slope of the inferior facet, convergence angle of the superior facet, circumference of the inferior articular facet, and circumference of the superior articular facet. Principal components analysis (varimax rotation) was used to determine sources of variation within the dataset. Multivariate analysis of variance and paired t tests were used to identify statistical differences between right and left sides for each variable. RESULTS: More than 99% of variance was accounted for across 11 eigenvectors, with most eigenvectors dominated by a single variable. Bilateral comparison of variables showed a significant difference between sides for the anteroposterior length of the superior facet, the width of the superior facet, the width of the vertebral canal, anteroposterior diameter of the inferior facet, and convergence angle of the superior facet (all significant at P < .05). CONCLUSION: Based on statistical analysis of the atlantal variables, vertebral canal width and the morphology of the superior and inferior articular facets showed significant asymmetry. The role of these asymmetries related to the biomechanics of the C0 through C1 and encroachment on the spinal cord warrant further investigation. PMID- 18394499 TI - Effects of myofascial release after high-intensity exercise: a randomized clinical trial. AB - OBJECTIVE: The usefulness of massage as a recovery method after high-intensity exercise has yet to be established. We aimed to investigate the effects of whole body massage on heart rate variability (HRV) and blood pressure (BP) after repeated high-intensity cycling exercise under controlled and standardized pretest conditions. METHODS: The study included 62 healthy active individuals. After baseline measurements, the subjects performed standardized warm-up exercises followed by three 30-second Wingate tests. After completing the exercise protocol, the subjects were randomly assigned to a massage (myofascial release) or placebo (sham treatment with disconnected ultrasound and magnetotherapy equipment) group for a 40-minute recovery period. Holter recording and BP measurements were taken after exercise protocol and after the intervention. RESULTS: After the exercise protocol, both groups showed a significant decrease in normal-to-normal interval, HRV index, diastolic BP (P > .001), and low-frequency domain values (P = .006). After the recovery period, HRV index (P = .42) and high-frequency (HF) (P = .94) values were similar to baseline levels in the massage group, whereas the HRV index tended (P = .05) to be lower and the HF was significantly (P < .01) lower vs baseline values in the placebo group, which also showed a tendency (P = .06) for HF to be lower than after the exercise. Likewise, diastolic BP returned to baseline levels in the massage group (P = .45) but remained lower in the placebo group (P = .02). CONCLUSION: Myofascial release massage favors the recovery of HRV and diastolic BP after high intensity exercise (3 Wingate tests) to preexercise levels. PMID- 18394500 TI - Survey of faulty postures and associated factors among Chinese adolescents. AB - OBJECTIVE: This study investigates the prevalence of common faulty postures among adolescents and identify if significant relationships existed among the number of faulty postures, psychologic distress, and musculoskeletal symptoms. METHODS: The Musculoskeletal Questionnaire and Chinese Health Questionnaire were randomly distributed to 300 high school students in the Tainan area. On-campus postural screening, which included digital photography, manual muscle tests, and flexibility tests, was also performed. RESULTS: Two hundred eighty-seven participants completed all of the evaluations. The most common faulty posture was uneven shoulder level (36%), followed by forward head posture (25%). There was a sex difference between groups. The incidence of forward head posture for the male students was higher than that of the female students (P < .0001). In addition, the high psychologic distress group tended to have a higher prevalence of uneven shoulder height than that of the low psychologic distress group (P < .0001). As for the correlation analysis, the researchers did not find a high correlation among the scores of the faulty posture, psychologic distress, and musculoskeletal symptoms. CONCLUSION: The results of this study show that the incidence of faulty posture was high for the adolescent group, especially for the uneven shoulder level. Subjects' awareness about being assessed might decrease the incidence for some faulty posture. However, the relationships among the number of faulty postures, psychologic distress, and musculoskeletal symptoms were low. We suggest that there are multiple factors that might contribute to musculoskeletal symptoms; faulty posture could be one important factor that causes symptoms. PMID- 18394501 TI - Effects of a manually assisted mechanical force on cutaneous temperature. AB - OBJECTIVE: Digitized infrared segmental thermometry (DIST) is a tool used for measuring cutaneous temperature (CT). This project ascertains the effect of a manually assisted mechanical force producing a chiropractic adjustment in the lumbar spine after the Activator Methods Chiropractic Technique on CT during 2 different time recording periods (TRPs). METHODS: Sixty-six healthy subjects (36 women and 30 men) without acute low back conditions or symptoms were recruited. Subjects were randomly divided into 2 groups based on the length of the acclimatization period (8 or 30 minutes; TRP(8) and TRP(30), respectively). In turn, each recording period group was divided into 3 subgroups (n = 11 per subgroup): treatment, sham, and control subgroups. Bilateral DIST was conducted at L-4 (TRP(30)) and L-5 (TRP(8)) using infrared cameras (Subluxation Station Insight 7000; Chiropractic Leadership Alliance, Mahwah, NJ). RESULTS: Before treatment (t(-0.5)), the TRP(8) CT was significantly different between the ipsilateral and the contralateral sides for all subgroups. At 10 minutes (t(10)) after intervention, CT increased significantly (P < .05) for the treatment group but not for the sham and control groups. In contrast, there were no significant differences in the TRP(30) CT before treatment between the ipsilateral and the contralateral sides; but at t(10), CT was significantly (P < .05) greater for all 3 subgroups compared with preintervention CT. CONCLUSION: Contacting the skin with the instrument with (treatment group TRP(30)) or without (sham group TRP(30)) a thrust with a sustained pressure stronger than the loading principle taught in the Activator Methods Chiropractic Technique protocol or a thrust respecting the standard loading principle (treatment group TRP(8)) of the instrument produced a CT cooling immediately after the adjustment. Furthermore, we observed that when contacting the skin with the instrument with a thrust respecting the standard loading principle (treatment group TRP(8)) of the instrument, it produced a secondary cooling at t(5) followed by a rewarming at t(10). Finally, contacting the skin with the instrument without a thrust and respecting the standard loading principle (sham TRP(8)) of the instrument did not produce a CT change. PMID- 18394502 TI - Prospective case series on the effects of lumbosacral manipulation on dysmenorrhea. AB - OBJECTIVE: The objective of this prospective case series was to collect preliminary data as to the effectiveness of a specific chiropractic technique, drop table method, in the treatment of primary dysmenorrhea. METHODS: Over a 4 week period, 16 females were screened for symptoms of primary dysmenorrhea and motion restrictions of the lumbosacral spine. Thirteen subjects were enrolled into the study. Bilateral and unilateral lumbosacral flexion and extension restrictions were treated using drop table manipulations 3 times during each of the 2 consecutive menstrual cycles. Before entering the study and at the end of each menstrual cycle, the subjects self-reported ratings of menstrual pain (abdominal, pelvic, and low back pain) and associated symptoms of primary dysmenorrhea using Numeric Pain Scale. Numeric Pain Scale ratings for menstrual pain were the primary outcome measures. RESULTS: The median age was 26 years, and the median self-reported duration of the symptoms was 12 years. At baseline, all subjects reported pain severity scores of 5 or higher for at least 2 of 3 anatomical sites: lower or general abdominal pain and/or lower back pain. Using the 95% confidence interval (CI) as an estimate, clinically meaningful changes (<5) in general abdominal pain and lower back pain were evident for most patients during the treatment phase, whereas for lower abdominal pain, the improvements were subject and cycle dependent. CONCLUSIONS: Menstrual pain associated with primary dysmenorrhea may be alleviated with treatment of motion segment restrictions of the lumbosacral spine with drop table technique. PMID- 18394503 TI - Fibular collateral ligament-biceps femoris calcific bursitis causing flexion contracture in the knee, external rotation in the leg, and equinus deformity in the ankle. AB - OBJECTIVE: The fibular collateral ligament (FCL)-biceps femoris (BF) bursa is not a commonly known location for calcific bursitis. A case of FCL-BF calcific bursitis is presented. CLINICAL FEATURES: An 18-year-old man had a blunt trauma on the lateral side of his right knee approximately 1 year before his examination. The pain and physical distortion developed in the trauma region over time. Physical examination revealed a 30 degrees flexion contracture and slight valgus deformity in the right knee. There was 30 degrees of external rotation in the leg and a 15 degrees equinus deformity in the ankle. The patient had toe-toe gait. There was a tender swelling 3 x 4 cm in size on the posterolateral aspect of the right knee. INTERVENTION AND OUTCOME: Plain radiography and computed tomography revealed a calcific mass, approximately 1.5 x 4 cm in size, between soft tissues adjacent to the posterolateral side of the right lateral femoral condyle. There were articular cartilage thinning and joint space narrowing at the lateral site of the knee joint, and magnetic resonance imaging showed bone marrow edema at the lateral condyles of the tibia and femur. The enlarged calcified bursa was excised. A segment of the contracted iliotibial tract was excised, and a fractional lengthening was performed on the BF tendon. CONCLUSION: This case report revealed that calcific bursitis can occur in the FCL-BF bursa, which may lead to complications when neglected. PMID- 18394505 TI - Changes in pressure pain sensitivity in latent myofascial trigger points in the upper trapezius muscle after a cervical spine manipulation in pain-free subjects. PMID- 18394506 TI - Resilience in aging: the real experts. PMID- 18394507 TI - The value of vitamin D(3) over vitamin D(2) in older persons. PMID- 18394508 TI - CMS clarification of survey and certification interpretive guidelines. PMID- 18394509 TI - Aging in place and negotiated risk agreements. AB - An assisted living community (i.e., residence [ALR]) that responds to a resident's changing needs by adjusting services and level of care criteria means that the resident is less likely to be discharged to a higher level of care, such as a nursing home, and will "age in place." A "managed" or negotiated risk agreement (NRA) can facilitate aging in place by accommodating a resident's choices and potential risks of remaining in the residence. Although more than half the states speak to aging-in-place in their assisted living regulations, far fewer provide guidelines for negotiated risk discussion and its influence on service plans. Managing decline and the risks to a resident's safety and well being can perhaps be addressed by "upstream palliative care," a concept and practice that respects a resident's quality of life interests while meeting and supporting their physical, mental, and emotional needs. This article discusses these issues as well as the capacity of a cognitively impaired/demented resident to enter into an NRA and its continuance when resident understanding is questionable, and dying-in-place. PMID- 18394510 TI - Moving from achievement to transformation. PMID- 18394511 TI - Social and cultural construction of urinary incontinence among Korean American elderly women. AB - The purpose of this article is to describe the social and cultural constructions that influence help-seeking for urinary incontinence (UI) among Korean American elderly women. Many Korean American elder women do not perceive UI as a problem worthy of attention. The reason may lie in its social and cultural construction. The social construction is rooted in the collectivist nature of Korean culture, which makes UI a family, rather than an individual, problem. The cultural construction is related to Confucianism, which directs conceptions about ageism, shame, and fatalism. This article will help the gerontological nurse to better understand Korean American elderly women's sociocultural background related to UI care and could lead to appropriate family centered interventions to manage or treat UI in this population. PMID- 18394512 TI - Views of African American Nursing Home residents about living wills. AB - Advance directives may be understood differently by various cultural and racial groups. These differences need to be identified so that individualized treatment decisions can be made according to patient preferences and communicated to clinicians. This exploratory, descriptive, qualitative research asked elderly African American nursing home residents why they had not completed a living will. Focus groups were conducted at 3 nursing homes and the participants spoke of: 1) not wanting artificial life-supporting treatments, 2) not being asked about living wills, 3) the importance of their families in the decision-making process, 4) the role of physicians in end-of-life care, and 5) having a living will was probably a good idea. Elderly nursing home residents would benefit from discussions within their family and cultural community about goals of therapy at the end of life that may lead to increased use of advanced directive documents. PMID- 18394513 TI - "Eye" dentifying vision impairment in the geriatric patient. AB - Vision impairment is highly prevalent yet undetected in the elderly population. Without screening and early detection age-related vision loss can lead to various adverse effects. The purpose of this review is to assess the methods of screening geriatric patients for visual impairment that can be employed by nurse practitioners in the primary care setting. A series of databases were searched utilizing the key words aged, geriatric, vision, vision tests, and screening. The Snellen chart, Pinhole assessment, Ophthalmoscope, Cardiff acuity test, Amsler grid, and subjective measures are discussed. All must be utilized cautiously because research has shown that these screening methods are limited in their sensitivity and specificity in detecting ocular disease. No combination of tools is ideal for identifying age-related vision loss although implications for practice are implied. PMID- 18394514 TI - Risk factors for mobility limitation in community-dwelling older adults: a social ecological perspective. AB - Although a variety of risk factors for mobility limitation in older adults have been examined, a collective review of relevant literature has not been reported. The purposes of this review are to report the intrapersonal, interpersonal, environmental, and organizational risk factors related to mobility limitation using a social ecological perspective and to discuss the direction of future clinical practice consistent with current literature on mobility limitation of community-dwelling older adults. Intrapersonal risk factors related to mobility limitation include advanced age, female gender, low socioeconomic status, comorbidity, lack of motivation (i.e., dependent personality, decreased self efficacy), lifestyle factors (i.e., sedentary lifestyle, smoking, obesity), and physiological factors (i.e., vitamin D deficiency, inflammation, poor nutritional status). Interpersonal risk factors related to mobility limitation include weak social networks and limited social activities. Geriatric clients may also experience a decline in mobility when they encounter environmental challenges such as an inconvenient home environment and lack of availability of services in their community, as well as lack of organizational resources stemming from social policy. Potential intervention strategies focused on modifiable risk factors may include lifestyle modifications, social networking programs, and enhancing awareness of environmental and organizational resources in the community for older adults at risk for mobility limitation. PMID- 18394515 TI - Perceived autonomy and self-care resources among senior center users. AB - Personal autonomy-the ability to make freely self-directed choices in one's life is considered critical to an older person's quality of life. Informed by model and role-modeling nursing theory, this descriptive, correlational study examined the relationship of perceived enactment of autonomy to self-care resources, both internal (age, race, functional ability) and external (social support, living arrangements, community-based service utilization, and service satisfaction) among 120 older adults from 6 Manhattan senior centers. Participants demonstrated positive perceived autonomy. Study participants used a variety of community-based services, and their satisfaction with the services was significantly linked to autonomy. Using multiple linear regression model, the only significant predictors of perceived autonomy were race (white > nonwhite), service satisfaction, and social support. Nursing strategies such as providing information that promotes informed decision making and encouraging independence, as well as working collaboratively with members of an older persons' social network, can enhance an older persons' autonomy. PMID- 18394516 TI - Robotics and neuroscience. Editorial. PMID- 18394517 TI - Does the brain make waves to improve stability? AB - In many ways, roboticians and the human brain are faced with the same problem: How does one control movement from a distance? In both cases, delays in the transmission of information play an important role, either because the distances to be covered are long (imagine controlling a robot arm on the moon from a command center on Earth), or because the underlying hardware is slow (nerves transmit information much more slowly than wires, radio waves or light). Delays have a debilitating effect on feedback control systems; causes and effects can bounce back and forth between distant sites, resulting in oscillatory behavior that can grow without bound. Control engineers have developed the concept of wave variables to combat this problem-by mimicking a flexible rod, wave variables constrain movement of the master and slave during the delay, ensuring stable overall behavior [G. Niemeyer, J.J.E. Slotine, Stable adaptive teleoperation, IEEE J. Ocean Eng. 16 (1991) 152-162; G. Niemeyer, J.J.E. Slotine, Toward bilateral internet teleoperation, in: Beyond Webcams, an Introduction to Online Robots, MIT Press, 2002]. Mother Nature may, however, deserve the patent on this solution. As we show here, the properties of nerves, muscles and sensory organs combine to form a natural wave variable control system that is immune to the problems of feedback delays. PMID- 18394518 TI - Contact forces evoked by transcranial magnetic stimulation of the motor cortex in a multi-finger grasp. AB - Transcranial magnetic stimulation (TMS) is a tool of choice to study the functionality of the corticospinal pathway. In this study, we used single-pulse TMS at different intensities and during different levels of grasping force, to stimulate the hand area of the left primary motor cortex (M1). We measured, the TMS-evoked forces, or motor evoked forces (MEFs) in a multi-fingered three-point grasp in addition of the conventional motor evoked potentials (MEPs) from the right forearm and intrinsic hand muscles. This paper aims at presenting the viability of this innovative approach and some preliminary results. The timing (i.e., latencies and peak times), amplitudes and directions of the MEF were analyzed. We found that the TMS evoked synergistic increases of the force magnitudes, akin to those observed when participants voluntarily increased the grip force. The MEF sizes and MEP amplitudes increased with TMS intensity in most cases. The grip force (which measures the overall force involved in the grasp) and the net force (which measures the net effect of all contact forces exerted on the object) seem to be differently affected by single TMS pulses of the motor cortex. PMID- 18394519 TI - Tactile flow explains haptic counterparts of common visual illusions. AB - Interaction with the external world requires the ability to perceive dynamic changes in complex sensorial input and react promptly. Here, we show that perception of dynamic stimuli in the visual and tactile sensory modalities share fundamental psychophysical aspects that can be explained by similar computational models. In vision, optic flow provides information on relative motion between the individual and the content of percept. For instance, radial patterns of optic flow are used to estimate time before contact with an approaching object [J.J. Gibson, What gives rise to the perception of motion? Psychol. Rev. 75 (1968) 335 346]. Similarly, in the tactile modality, radial patterns of stimuli provide information on softness of probed objects [A. Bicchi, D. De Rossi, E.P. Scilingo, The role of the contact area spread rate (CASR) in haptic discrimination of softness, IEEE Trans. Rob. Autom. 16 (2000) 496-504]. Optic flow is also invoked to explain several visual illusions, including the well-known "barber-pole" effect [N. Fisher, J.M. Zanker, The directional tuning of the barber-pole illusion, Perception 30 (2001) 1321-1336]. Here, we introduce a computational model of tactile flow, which is intimately related to existing models for the visual counterpart. The model accounts for psychophysical aspects of dynamic tactile perception and predicts illusory phenomena in the tactile domain, analogous to the barber-pole effect. When subjects touched translating pads with differently oriented gratings, they perceived a direction of motion that was significantly biased towards the orientation of the gratings. Therefore, these findings indicate that visual and tactile flow share similarities at the psychophysical and computational level and may be intended for similar perceptive goals. Results of this analysis have impact on the engineering of better haptic and multimodal interfaces for human-computer interaction. PMID- 18394520 TI - A brief taxonomy of tactile illusions and demonstrations that can be done in a hardware store. AB - This paper surveys more than twenty types of tactile illusions and discusses several of their aspects. These aspects include the ease with which they can be demonstrated and whether they have clear visual analogs. The paper also shows how to construct equipment made of simple supplies able to deliver well-controlled tactile signals in order to conveniently demonstrate four different tactile illusions. PMID- 18394521 TI - Tri-modal integration of visual, tactile and auditory signals for the perception of sequences of events. AB - We investigated the interactions between visual, tactile and auditory sensory signals for the perception of sequences of events. Sequences of flashes, taps and beeps were presented simultaneously. For each session, subjects were instructed to count the number of events presented in one modality (Target) and to ignore the stimuli presented in the other modalities (Background). The number of events presented in the background sequence could differ from the number of events in the target sequence. For each session, we quantified the Background-evoked bias by comparing subjects' responses with and without Background (Target presented alone). Nine combinations between vision, touch and audition were tested. In each session but two, the Background significantly biased the Target. Vision was the most susceptible to Background-evoked bias and the least efficient in biasing the other two modalities. By contrast, audition was the least susceptible to Background-evoked bias and the most efficient in biasing the other two modalities. These differences were strongly correlated to the relative reliability of each modality. In line with this, the evoked biases were larger when the Background consisted of two instead of only one modality. These results show that for the perception of sequences of events: (1) vision, touch and audition are automatically integrated; (2) the respective contributions of the three modalities to the integrated percept differ; (3) the relative contribution of each modality depends on its relative reliability (1/variability); (4) task irrelevant stimuli have more weight when presented in two rather than only one modality. PMID- 18394522 TI - Backward and common-onset masking of vibrotactile stimuli. AB - To inform the design of haptic information displays for noisy environments, we investigated two mechanisms for temporal masking of vibrotactile stimuli (backwards and common-onset) using a commodity display. We used a two-channel setup, presenting stimuli to the middle and ring finger of a participant's right hand. The stimuli consisted of 250 Hz sinusoidal waveforms displayed at a fixed amplitude in various combinations of duration (0, 30 or 300 ms) and stimulus onset asynchrony (0 or 30 ms). In anticipation of future embedded applications where signals are deliberately masked but levels cannot be individualized, signals were standardized at conservative (harder to mask) levels. Our results confirm the existence of a statistically significant masking effect for both forms of haptic masking explored, with common-onset exhibiting a significantly larger masking effect than backwards. However, an analysis of confidence in response levels shows no difference between the two successful masking techniques. We discuss mechanisms that could be responsible for these results, which have implications for the design of user interfaces that rely on tactile transmission of information. PMID- 18394523 TI - Temporal prediction of touch instant during observation of human and robot grasping. AB - The aim of the present work was to test the ability to predict the instant at which a grasping hand touches an object. Our hypothesis was that, because of the activation of the mirror-neuron system, the same predictive process necessary for action execution should be active during observation. Experimental evidence indicates, however, that not only observed actions but also observed objects automatically activate observer's motor repertoire. What happens, therefore, if the observed action is different from the one automatically evoked by the vision of the object? To answer this question we presented subjects with two different grasping actions: the one most suitable for the presented object and a less appropriate one. Subjects were required to detect the instant at which the demonstrator's hand touched the object. In a further condition, subjects were required to detect the outcome of an action performed by a robotic arm moving with constant kinematics. Results showed that while in the case of robot grasping subjects responded before the touch instant, in the case of human grasping the response followed the touch instant, but occurred much earlier than simple reaction times. This demonstrates that subjects were able to predict the outcome of the seen action. The predictive capability was specifically enhanced during observation of the "suitable" grasping. We interpret these results as an indication of the synergic contribution of both object-related (canonical) and action-related (mirror) neurons during observation of actions directed towards graspable objects. PMID- 18394524 TI - From self-observation to imitation: visuomotor association on a robotic hand. AB - Being at the crux of human cognition and behaviour, imitation has become the target of investigations ranging from experimental psychology and neurophysiology to computational sciences and robotics. It is often assumed that the imitation is innate, but it has more recently been argued, both theoretically and experimentally, that basic forms of imitation could emerge as a result of self observation. Here, we tested this proposal on a realistic experimental platform, comprising an associative network linking a 16 degrees of freedom robotic hand and a simple visual system. We report that this minimal visuomotor association is sufficient to bootstrap basic imitation. Our results indicate that crucial features of human imitation, such as generalization to new actions, may emerge from a connectionist associative network. Therefore, we suggest that a behaviour as complex as imitation could be, at the neuronal level, founded on basic mechanisms of associative learning, a notion supported by a recent proposal on the developmental origin of mirror neurons. Our approach can be applied to the development of realistic cognitive architectures for humanoid robots as well as to shed new light on the cognitive processes at play in early human cognitive development. PMID- 18394525 TI - Bio-inspired sensorization of a biomechatronic robot hand for the grasp-and-lift task. AB - It has been concluded from numerous neurophysiological studies that humans rely on detecting discrete mechanical events that occur when grasping, lifting and replacing an object, i.e., during a prototypical manipulation task. Such events represent transitions between phases of the evolving manipulation task such as object contact, lift-off, etc., and appear to provide critical information required for the sequential control of the task as well as for corrections and parameterization of the task. We have sensorized a biomechatronic anthropomorphic hand with the goal to detect such mechanical transients. The developed sensors were designed to specifically provide the information about task-relevant discrete events rather than to mimic their biological counterparts. To accomplish this we have developed (1) a contact sensor that can be applied to the surface of the robotic fingers and that show a sensitivity to indentation and a spatial resolution comparable to that of the human glabrous skin, and (2) a sensitive low noise three-axial force sensor that was embedded in the robotic fingertips and showed a frequency response covering the range observed in biological tactile sensors. We describe the design and fabrication of these sensors, their sensory properties and show representative recordings from the sensors during grasp-and lift tasks. We show how the combined use of the two sensors is able to provide information about crucial mechanical events during such tasks. We discuss the importance of the sensorized hand as a test bed for low-level grasp controllers and for the development of functional sensory feedback from prosthetic devices. PMID- 18394526 TI - Non-invasive brain-computer interface system: towards its application as assistive technology. AB - The quality of life of people suffering from severe motor disabilities can benefit from the use of current assistive technology capable of ameliorating communication, house-environment management and mobility, according to the user's residual motor abilities. Brain-computer interfaces (BCIs) are systems that can translate brain activity into signals that control external devices. Thus they can represent the only technology for severely paralyzed patients to increase or maintain their communication and control options. Here we report on a pilot study in which a system was implemented and validated to allow disabled persons to improve or recover their mobility (directly or by emulation) and communication within the surrounding environment. The system is based on a software controller that offers to the user a communication interface that is matched with the individual's residual motor abilities. Patients (n=14) with severe motor disabilities due to progressive neurodegenerative disorders were trained to use the system prototype under a rehabilitation program carried out in a house-like furnished space. All users utilized regular assistive control options (e.g., microswitches or head trackers). In addition, four subjects learned to operate the system by means of a non-invasive EEG-based BCI. This system was controlled by the subjects' voluntary modulations of EEG sensorimotor rhythms recorded on the scalp; this skill was learnt even though the subjects have not had control over their limbs for a long time. We conclude that such a prototype system, which integrates several different assistive technologies including a BCI system, can potentially facilitate the translation from pre-clinical demonstrations to a clinical useful BCI. PMID- 18394527 TI - Visual feedback distortion in a robotic environment for hand rehabilitation. AB - Robotic therapy offers a means of enhancing rehabilitation for individuals with chronic stroke or traumatic brain injury. The present research targets members of this population who demonstrate learned nonuse, a tendency to use affected limbs below the level of the individual's true capability. These individuals may not strive for difficult goals in therapy, which ultimately hampers their progress and the outcome of rehabilitation. Our research uses a paradigm called visual feedback distortion in which the visual feedback corresponding to force or distance is gradually changed by an imperceptible amount to encourage improved performance. Our first set of experiments was designed to assess the limits of imperceptible distortion for visual feedback concerning the force exerted or the distance moved by the index finger. A second set of experiments used these limits to gradually distort visual feedback in order to manipulate a subject's force or distance response. Based on this work, we designed a paradigm applying visual feedback distortion to the rehabilitation of individuals with chronic stroke and traumatic brain injury. Initial tests are reported for two subjects who participated in a six-week rehabilitation protocol. Each patient followed visual feedback distortion to levels of performance above that predicted by her performance during an initial assessment. Both patients showed functional improvements after participating in the study. Visual feedback distortion may provide a way to help a patient move beyond his or her self-assessed "best" performance, improving the outcome of robotic rehabilitation. PMID- 18394529 TI - Sonographers' communication: a view from the other side of health care. PMID- 18394530 TI - Strain without pain: application of parametric imaging of strain rate response for the quantitation of stress echocardiography. PMID- 18394532 TI - Tragedy of conducting a clinical trial; generic alert system needed. AB - Stopping a clinical trial without reaching the final objective is not the ideal outcome any researcher wants; sometimes ceasing is inevitable. Due to marginal inclusion of patients we were forced to cease our randomized clinical trial on the effectiveness of proprioceptive training on the development of chronic whiplash complaints a year after the start. Although incidence figures demonstrate that recruitment of the planned number of whiplash patients would be easily feasible, we were unable to enroll the amount of subjects. Several motives can be proposed that would have prevented this obliged halting from happening. Other studies also report impracticability of the planned number of whiplash injury patients. PMID- 18394533 TI - Why would physicians undervalue reviews by the Cochrane Collaboration? PMID- 18394534 TI - Few systematic reviews exist documenting the extent of bias: a systematic review. AB - OBJECTIVE: To summarize the evidence concerning bias and confounding in conducting systematic reviews (SRs). STUDY DESIGN AND SETTING: Literature was identified through searching the Cochrane Library, MEDLINE, PsycINFO until November 2006, and the authors' files. Studies were included if they were SRs of bias that can occur while conducting a SR. Risk of bias in the SRs was appraised using the Oxman and Guyatt index. RESULTS: Ten SRs were included. All examined biases related to searching for evidence (e.g., publication bias). One also reported bias associated with obtaining data from included studies (e.g., outcome reporting bias). To minimize bias, data suggest including unpublished material, hand searching for additional material, searching multiple databases, assessing for publication bias, and periodically updating SRs. No SRs were found examining biases related to choosing studies for inclusion or combining studies. CONCLUSIONS: There is little evidence from SRs to support commonly practiced methods for conducting SRs. No SRs summarized studies with prospective designs and most had moderate or minimal risk of bias. Future research should examine bias that can occur during the selection of studies for inclusion and the synthesis of studies, as well as systematically review the existing empirical evidence. PMID- 18394535 TI - The account for provider and center effects in multicenter interventional and surgical randomized controlled trials is in need of improvement: a review. AB - OBJECTIVE: To systematically review the account of center and provider effects in large surgical and interventional randomized controlled trials. STUDY DESIGN: A systematic review of multicenter interventional randomized trials of 200+ patients. The search included Medline from 1 January 2000 through 11 October 2005 and a hand search of the bibliographies of retrieved articles. One author abstracted all data using standardized abstraction forms; a second reviewer assessed a random sample of reports as quality-assurance procedure. RESULTS: Sixty-eight reports met inclusion criteria. The trials predominantly reported on cardiology (n=23, 34%). The number of participating providers was reported in 11 trials (16%). Both the performed control and performed interventional procedures were described in 43 trials (63%). The use of stratified random allocation on center was reported in 26 trials (38%) and on provider in 6 trials (9%). The analysis was adjusted for center in four trials (6%) and for provider in three trials (4%). CONCLUSIONS: Only few trials account for center or provider effect in the design and analysis. Authors and journal editors could play an important role in improving the reporting of trials. PMID- 18394536 TI - Poor reporting and inadequate searches were apparent in systematic reviews of adverse effects. AB - OBJECTIVE: Systematic reviews incorporating adverse effects are assuming increasing importance as questions raised extend beyond clinical effectiveness to all effects (beneficial and harmful). The aim of this study was to survey the methods used to identify relevant studies for systematic reviews of adverse effects. STUDY DESIGN AND SETTING: All records within the Database of Abstracts of Reviews of Effects and the Cochrane Database of Systematic Reviews were scanned for systematic reviews in which the primary outcomes were adverse effects. Two information professionals independently assessed the methods used to identify relevant research as reported in the 277 reviews that met the inclusion criteria. RESULTS: A major weakness of the reviews was inadequate reporting of the search strategies used. In addition, of the reviews that did report a search strategy, few used the sensitive search strategies recommended for systematic reviews. The majority of reviews did not search more than one or two databases, and few other methods of identifying information were used. CONCLUSION: This investigation shows the variation in the searching element of systematic reviews of adverse effects and demonstrates that the reporting of the methods used to identify research in such reviews could be vastly improved. PMID- 18394537 TI - Systematic reviews and original articles differ in relevance, novelty, and use in an evidence-based service for physicians: PLUS project. AB - OBJECTIVES: To describe the ratings from physicians, and use by physicians, of high quality, clinically pertinent original articles and systematic reviews from over 110 clinical journals and the Cochrane Database of Systematic Reviews (CDSRs). STUDY DESIGN AND SETTING: Prospective observational study. Data were collected via an online clinical rating system of relevance and newsworthiness for quality-filtered clinical articles and via an online delivery service for practicing physicians, during the course of the McMaster Premium LiteratUre Service Trial. Clinical ratings of articles in the MORE system by over 1,900 physicians were compared and the usage rates over 13 months of these articles by physicians, who were not raters, were examined. RESULTS: Systematic reviews were rated significantly higher than original articles for relevance (P<0.001), but significantly lower for newsworthiness (P<0.001). Reviews published in the CDSR had significantly lower ratings for both relevance (P<0.001) and newsworthiness (P<0.001) than reviews published in other journals. Participants accessed reviews more often than original articles (P<0.001), and accessed reviews from journals more often than from CDSR (P<0.001). CONCLUSION: Physician ratings and the use of high-quality original articles and systematic reviews differed, generally favoring systematic reviews over original articles. Reviews published in journals were rated higher and accessed more often than Cochrane reviews. PMID- 18394538 TI - Adjusted indirect comparison may be less biased than direct comparison for evaluating new pharmaceutical interventions. AB - OBJECTIVE: To investigate discrepancies between direct comparison and adjusted indirect comparison in meta-analyses of new versus conventional pharmaceutical interventions. STUDY DESIGN AND SETTING: Results of direct comparison were compared with results of adjusted indirect comparison in three meta-analyses of new versus conventional drugs. The three case studies are (1) bupropion versus nicotine replacement therapy for smoking cessation, (2) risperidone versus haloperidol for schizophrenia, and (3) fluoxetine versus imipramine for depressive disorders. RESULTS: In all the three cases, effects of new drugs estimated by head-to-head trials tend to be greater than that by adjusted indirect comparisons. The observed discrepancies could not be satisfactorily explained by the play of chance or by bias and heterogeneity in adjusted indirect comparison. This observation, along with analysis of possible systematic bias in the direct comparisons, suggested that the indirect method might have produced less biased results. Simulations found that adjusted indirect comparison may counterbalance bias under certain circumstances. CONCLUSION: Adjusted indirect comparison could be used to cross-examine the validity and applicability of results from head-to-head randomized trials. The hypothesis that adjusted indirect comparison may provide less biased results than head-to-head randomized trials needs to be investigated by further research. PMID- 18394539 TI - Falsified papers in high-impact journals were slow to retract and indistinguishable from nonfraudulent papers. AB - OBJECTIVE: The aim was to evaluate papers retracted due to falsification in high impact journals. STUDY DESIGN AND SETTING: We selected articles retracted due to allegations of falsification in January 1, 1980 to March 1, 2006 from journals with impact factor >10 and >30,000 annual citations. We evaluated characteristics of these papers and misconduct-involved authors and assessed whether they correlated with time to retraction. We also compared retracted articles vs. matched nonretracted articles in the same journals. RESULTS: Fourteen eligible journals had 63 eligible retracted articles. Median time from publication to retraction was 28 months; it was 79 months for articles where a senior researcher was implicated in the misconduct vs. 22 months when junior researchers were implicated (log-rank P<0.001). For the 25 implicated authors, the median time from the first publication of a fraudulent paper to the first retraction was 34 months, again with a clear difference according to researcher rank (log-rank P=0.001). Retracted articles didn't differ from matched nonretracted papers in citations received within 12 months, number of authors, country, funding, or field, but were twofold more likely to have multinational authorship (P=0.049). CONCLUSIONS: Retractions due to falsification can take a long time, especially when senior researchers are implicated. Fraudulent articles are not obviously distinguishable from nonfraudulent ones. PMID- 18394540 TI - Oncology employees valued patient-oriented research over quality of work-life research: a qualitative study. AB - OBJECTIVES: To assess the role of quality of work-life (QWL) research in a clinical research environment. STUDY DESIGN AND SETTING: A phenomenological qualitative research study was conducted. This research was conducted in a large Canadian ambulatory cancer treatment center that serves over 2.3 million people in Ontario, Canada. A variety of research activities are carried out at the center including clinical trials and a QWL project. PARTICIPANTS: 32 employees from a variety of activity groups were recruited by electronic mail and interviewed for approximately 1 hour. Two main questions ("Tell me about the research at the center"; "What do you think about the research activities at the center?") were posed to participants. RESULTS: There was a perceived status difference in the research conducted at the center. PARTICIPANTS appeared to value clinical research (patient interests) over the QWL research (their own interests). Consistent with this perceived status difference, participants' perceptions and descriptions of research were framed in the context of clinical research. CONCLUSION: Conducting workplace research in a clinical setting may pose challenges to occupational researchers. However, allowing oncology employees to participate in clinical research may enhance their QWL. PMID- 18394541 TI - A parametric analysis of ordinal quality-of-life data can lead to erroneous results. AB - OBJECTIVE: Measurements from health-related quality-of-life (HRQoL) studies, although usually of an ordered categorical nature, are typically treated as continuous variables, allowing the calculation of mean values and the administration of parametric statistics, such as t-tests. We investigated whether parametric, compared to nonparametric, analyses of ordered categorical data may lead to different conclusions. STUDY DESIGN AND SETTING: HRQoL data were obtained from patients with a diagnosis of asthma (n=192) and chronic obstructive pulmonary disease (COPD; n=88) at two time points. The impact of the group factor (asthma vs. COPD) and the time factor (t1 vs. t2) on HRQoL was analyzed with a metric approach (repeated measures ANOVA) and two ordinal approaches (each with a nonparametric repeated measures ANOVA). RESULTS: Using the metric approach, a significant effect of "group" (P=0.0061) and "time" (P=0.0049) on HRQoL was found. The first ordinal approach (ranked total score) still showed a significant effect for "group" (P=0.0033) with a worse HRQoL for patients suffering from COPD. In the second approach (ranks for each HRQoL item and summed ranks), there were no significant effects. CONCLUSION: Applying simple parametric methods to ordered categorical HRQoL scores led to different results from those obtained with nonparametric methods. In these cases, an ordinal approach will prevent inappropriate conclusions. PMID- 18394542 TI - Lysholm scale and WOMAC index were responsive in prospective cohort of young general practice patients. AB - OBJECTIVE: To determine the construct validity and responsiveness of the Lysholm knee scoring scale and the WOMAC osteoarthritis index in adolescents and young adults with knee complaints in general practice. STUDY DESIGN AND SETTING: In the framework of a prospective cohort study with 1-year follow-up, we included 314 patients aged 12-35 years consulting the general practitioner for incident knee complaints. Subgroup analyses of traumatic and nontraumatic knee complaints and of adolescents and adults were performed. RESULTS: Construct validity was adequate for both questionnaires both in traumatic and nontraumatic patients (aged 12-35) and in adolescents (12-17) and young adults (18-35). Effect size (ES) and standardized response mean (SRM) for both Lysholm and WOMAC global scores were moderate in nontraumatic patients and high in traumatic patients. Guyatt's responsiveness statistic was high in both subpopulations. Adolescents showed high responsiveness with all measures on the Lysholm scale, and moderate (Guyatt's statistic) to high responsiveness (ES and SRM) on the WOMAC index. Young adults showed high responsiveness with all measures on both instruments. CONCLUSION: Although neither of the scales was developed for use in adolescents and young adults in general practice, both scales show adequate responsiveness, content, and construct validity in this population. PMID- 18394543 TI - The short-form chronic respiratory disease questionnaire was a valid, reliable, and responsive quality-of-life instrument in acute exacerbations of chronic obstructive pulmonary disease. AB - OBJECTIVE: To assess the psychometric properties of the short-form chronic respiratory disease questionnaire (SF-CRQ) as a quality-of-life (QOL) instrument in chronic obstructive pulmonary disease (COPD) exacerbations. STUDY DESIGN AND SETTING: In a prospective multicenter cohort study, consecutive emergency department (ED) patients with COPD exacerbation were interviewed using the SF-CRQ and other instruments. Baseline information was collected in the ED and from follow-up data 2 weeks later. The results of the SF-CRQ were compared with the results of the other instruments and clinical variables by correlation and factor analyses. RESULTS: Of the 301 enrolled patients, 80% reported improvements across each of the domains of the SF-CRQ over the 2-week post-ED period. Overall median changes for the dyspnea, fatigue, emotional function, and mastery domains were 2, 1, 1, and 1.5, respectively (P<0.001 for each domain). Correlation and factor analyses support their convergent/divergent validity and construct validity. The reliability for the change score of the SF-CRQ was high (Cronbach's alpha coefficient, 0.82). Overall minimal clinically important difference for improvement in the SF-CRQ was 1.01 (95% confidence interval, 0.72-1.31). CONCLUSION: SF-CRQ is a valid, reliable, and responsive instrument for the assessment of short-term QOL change in patients with COPD exacerbations. PMID- 18394544 TI - Self-reported information on the diagnosis of colorectal cancer was reliable but not necessarily valid. AB - OBJECTIVE: Self-report is commonly used in epidemiologic studies; however, few data exist on the reliability and validity of this method for eliciting information related to the diagnosis of colorectal cancer. We examined the test retest reliability and validity of colorectal cancer patients reporting on the process of their diagnosis. STUDY DESIGN AND SETTING: One hundred and sixteen participants completed two telephone interviews, 1 month apart, and 95 general practitioners (GPs) completed a written questionnaire, to elicit information relating to key elements of the process of diagnosis of colorectal cancer. RESULTS: Acute symptoms such as rectal bleeding had higher reliability and validity than more general symptoms. Colonoscopy was the most accurately recalled diagnostic test. Recall of diagnosis date, and date of colonoscopy, had high test retest reliability. There were considerable differences between dates of diagnostic tests given by participants and GPs, but there was no evidence of a bias in a particular direction. Accuracy of recall did not diminish as time from diagnosis increased. CONCLUSION: This study confirms that self-reported symptoms, tests, and dates in the colorectal cancer diagnostic pathway are generally reliable; however, the validity of reported symptoms and tests can be moderate to poor. PMID- 18394545 TI - Qualiveen, a urinary-disorder specific instrument: 0.5 corresponds to the minimal important difference. AB - OBJECTIVE: This study assessed the interpretability of changes in Qualiveen, a specific health-related quality-of-life (HRQOL) questionnaire for urinary disorders (UD) in multiple sclerosis (MS) patients. Qualiveen has 30 items focusing on four aspects of patients' lives related to their urinary problems: bother with limitations (nine items), frequency of limitations (eight items), fears (eight items), and feelings (five items). Response options are framed as 5 point Likert-type scales, and scores on each domain range from 0 to 4. STUDY DESIGN AND SETTING: One hundred twenty-one MS patients with UD completed Qualiveen at baseline and 2-10 weeks later. At the second visit, patients also made global ratings of change in urinary HRQOL (GRC) with a 15-point scale from 7 to +7. We addressed the validity of GRC by examining its relationships with pre and post-Qualiveen scores and determined the mean change in score on each Qualiveen domain associated with small, moderate, and large changes in the corresponding GRC. RESULTS: The correlation of the change score with the corresponding GRC was substantially stronger than the correlation with either pre or post-Qualiveen score, establishing the validity of the GRC. In all domains, a within-patient change in Qualiveen score of approximately 0.5 (range: 0.36-0.72) represents the minimal important difference. A change in score of approximately 0.7 corresponds to a moderate change in quality of life (range: 0.55-0.86). CONCLUSIONS: These estimates will help in interpreting the results of subsequent studies that use Qualiveen to examine the impact of interventions designed to improve urinary-specific HRQOL. PMID- 18394546 TI - Comorbidity information in older patients at an emergency visit: self-report vs. administrative data had poor agreement but similar predictive validity. AB - OBJECTIVES: The objectives of this study were (1) to ascertain the level of agreement between the Charlson Comorbidity Index (CCI) based on self-report vs. administrative records, and factors affecting agreement and (2) to compare the predictive validity of the two indices in a sample of older emergency department (ED) patients. STUDY DESIGN AND SETTING: The study was a secondary analysis of data from a randomized trial of an ED-based intervention. The self-report and administrative CCI were compared using the intraclass correlation coefficient (ICC). Factors examined for effect on agreement included health service utilization, age, and sex. The predictive validity of the indices was compared using subsequent health services utilization and functional decline as outcomes. Participants (n=520) were recruited at four university-affiliated Montreal hospitals. Eligibility criteria included 65 years of age or older, able to speak English or French, and discharged to the community. RESULTS: Agreement between the two sources was poor to fair (overall weighted ICC 0.43 [95% confidence interval [CI]: 0.40, 0.47]). The predictive validity was similar for the two indices (area under the receiver-operating characteristic curve 0.51-0.66, depending on the outcomes). CONCLUSION: Agreement between self-report and administrative comorbidity data is only poor to fair but both have comparable predictive validity. PMID- 18394547 TI - Studying the causes of adverse drug reactions: circumvent the warning-interfered bias using the case-case comparison approach. PMID- 18394548 TI - Maximization of the sum of sensitivity and specificity as a diagnostic cutpoint criterion. PMID- 18394549 TI - C/EBPalpha in leukemogenesis: a matter of being in the right place with the right signals. AB - Leukemia-initiating cells can originate from hematopoietic progenitor cells that have acquired self-renewal capacity upon transformation with leukemic fusion genes. In this issue of Cancer Cell, Kirstetter and colleagues describe a mouse model for the frequent CEBPA mutations in human acute myeloid leukemia that result in the synthesis of only the 30kDa isoform, but not the 42kDa isoform of C/EBPalpha. This mutation uncouples C/EBPalpha's roles in myeloid differentiation and proliferation control. Furthermore, this mutation activates self-renewal in committed myeloid progenitor cells and induces myeloid malignancy with complete penetrance that is sustained by leukemia-initiating cells with a committed myeloid molecular signature. PMID- 18394550 TI - A molecular view of anti-ErbB monoclonal antibody therapy. AB - Abnormal activation of the epidermal growth factor receptor (EGFR) and its homolog HER2 (Neu/ErbB2) has been associated with many human cancers, and monoclonal antibodies targeting EGFR and HER2 are effective anticancer therapies. Structural studies of these receptors and antibodies have revealed much about how they function. In this issue of Cancer Cell, Schmiedel et al. report structural and functional studies of the anti-EGFR monoclonal antibody Matuzumab. They show that Matuzumab binds and inhibits EGFR in a manner distinctive from that of other therapeutic anti-EGFR antibodies and suggest that combination therapies with Matuzumab and other antibodies may prove beneficial. PMID- 18394551 TI - RanBP2: a tumor suppressor with a new twist on TopoII, SUMO, and centromeres. AB - In vertebrate cells, the small ubiquitin-like modifier SUMO plays a poorly defined role in targeting DNA topoisomerase II (TopoII) to centromeres (CENs) during mitosis, presumably to facilitate the untangling of sister chromatids as cells transition into anaphase. A new study by Dawlaty in the April 4 issue of Cell identifies the nucleoporin RanBP2 as a novel tumor suppressor that acts as a SUMO ligase for TopoII. Analysis of this interaction reveals TopoII recruitment to CENs is likely to play an important role in preventing chromosome segregation errors that lead to cancer. PMID- 18394552 TI - VHL inactivation: a new road to senescence. AB - Loss of the VHL tumor suppressor gene promotes cancer in several ways, including activation of the HIF transcription factors. HIF overexpression is associated with increased malignancy in many tumor types. So why is the spectrum of tumors associated with VHL loss restricted only to a few specific organs? In a recent paper in the March issue of Nature Cell Biology, Kaelin and colleagues provide a possible explanation, suggesting that VHL loss can also trigger senescence, a potent tumor suppressor mechanism. PMID- 18394553 TI - Modeling of C/EBPalpha mutant acute myeloid leukemia reveals a common expression signature of committed myeloid leukemia-initiating cells. AB - Mutations in the CEBPA gene are present in 7%-10% of human patients with acute myeloid leukemia (AML). However, no genetic models exist that demonstrate their etiological relevance. To mimic the most common mutations affecting CEBPA-that is, those leading to loss of the 42 kDa C/EBPalpha isoform (p42) while retaining the 30kDa isoform (p30)-we modified the mouse Cebpa locus to express only p30. p30 supported the formation of granulocyte-macrophage progenitors. However, p42 was required for control of myeloid progenitor proliferation, and p42-deficient mice developed AML with complete penetrance. p42-deficient leukemia could be transferred by a Mac1+c-Kit+ population that gave rise only to myeloid cells in recipient mice. Expression profiling of this population against normal Mac1+c Kit+ progenitors revealed a signature shared with MLL-AF9-transformed AML. PMID- 18394554 TI - Efficacy of TG101348, a selective JAK2 inhibitor, in treatment of a murine model of JAK2V617F-induced polycythemia vera. AB - We report that TG101348, a selective small-molecule inhibitor of JAK2 with an in vitro IC50 of approximately 3 nM, shows therapeutic efficacy in a murine model of myeloproliferative disease induced by the JAK2V617F mutation. In treated animals, there was a statistically significant reduction in hematocrit and leukocyte count, a dose-dependent reduction/elimination of extramedullary hematopoiesis, and, at least in some instances, evidence for attenuation of myelofibrosis. There were no apparent toxicities and no effect on T cell number. In vivo responses were correlated with surrogate endpoints, including reduction/elimination of JAK2V617F disease burden assessed by quantitative genomic PCR, suppression of endogenous erythroid colony formation, and in vivo inhibition of JAK-STAT signal transduction as assessed by flow cytometric measurement of phosphorylated Stat5. PMID- 18394555 TI - Selective inhibition of JAK2-driven erythroid differentiation of polycythemia vera progenitors. AB - Polycythemia Vera (PV) is a myeloproliferative disorder (MPD) that is commonly characterized by mutant JAK2 (JAK2V617F) signaling, erythrocyte overproduction, and a propensity for thrombosis, progression to myelofibrosis, or acute leukemia. In this study, JAK2V617F expression by human hematopoietic progenitors promoted erythroid colony formation and erythroid engraftment in a bioluminescent xenogeneic immunocompromised mouse transplantation model. A selective JAK2 inhibitor, TG101348 (300 nM), significantly inhibited JAK2V617F+ progenitor derived colony formation as well as engraftment (120 mg/kg) in xenogeneic transplantation studies. TG101348 treatment decreased GATA-1 expression, which is associated with erythroid-skewing of JAK2V617F+ progenitor differentiation, and inhibited STAT5 as well as GATA S310 phosphorylation. Thus, TG101348 may be an effective inhibitor of JAK2V617F+ MPDs in clinical trials. PMID- 18394556 TI - Blocking neuropilin-2 function inhibits tumor cell metastasis. AB - Metastasis, which commonly uses lymphatics, accounts for much of the mortality associated with cancer. The vascular endothelial growth factor (VEGF)-C coreceptor, neuropilin-2 (Nrp2), modulates but is not necessary for developmental lymphangiogenesis, and its significance for metastasis is unknown. An antibody to Nrp2 that blocks VEGFC binding disrupts VEGFC-induced lymphatic endothelial cell migration, but not proliferation, in part independently of VEGF receptor activation. It does not affect established lymphatics in normal adult mice but reduces tumoral lymphangiogenesis and, importantly, functional lymphatics associated with tumors. It also reduces metastasis to sentinel lymph nodes and distant organs, apparently by delaying the departure of tumor cells from the primary tumor. Our results demonstrate that Nrp2, which was originally identified as an axon-guidance receptor, is an attractive target for modulating metastasis. PMID- 18394557 TI - The signaling adaptor p62 is an important NF-kappaB mediator in tumorigenesis. AB - The balance between cell death and survival, two critical aspects of oncogenic transformation, determines the outcome of tumorigenesis. Nuclear factor-kappaB (NF-kappaB) is a critical regulator of survival; it is induced by the oncogene Ras and, when inhibited, accounts for the cell death response of Ras-transformed cells. Here, we show that the signaling adaptor p62 is induced by Ras, its levels are increased in human tumors, and it is required for Ras-induced survival and transformation. p62-/- mice are resistant to Ras-induced lung adenocarcinomas. p62 is necessary for Ras to trigger IkappaB kinase (IKK) through the polyubiquitination of tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6), and its deficiency produces increased reactive oxygen species (ROS) levels, which account for the enhanced cell death and reduced tumorigenicity of Ras in the absence of p62. PMID- 18394558 TI - Feedback circuit among INK4 tumor suppressors constrains human glioblastoma development. AB - We have developed a nonheuristic genome topography scan (GTS) algorithm to characterize the patterns of genomic alterations in human glioblastoma (GBM), identifying frequent p18(INK4C) and p16(INK4A) codeletion. Functional reconstitution of p18(INK4C) in GBM cells null for both p16(INK4A) and p18(INK4C) resulted in impaired cell-cycle progression and tumorigenic potential. Conversely, RNAi-mediated depletion of p18(INK4C) in p16(INK4A)-deficient primary astrocytes or established GBM cells enhanced tumorigenicity in vitro and in vivo. Furthermore, acute suppression of p16(INK4A) in primary astrocytes induced a concomitant increase in p18(INK4C). Together, these findings uncover a feedback regulatory circuit in the astrocytic lineage and demonstrate a bona fide tumor suppressor role for p18(INK4C) in human GBM wherein it functions cooperatively with other INK4 family members to constrain inappropriate proliferation. PMID- 18394559 TI - Matuzumab binding to EGFR prevents the conformational rearrangement required for dimerization. AB - An increasing number of therapeutic antibodies targeting tumors that express the epidermal growth factor receptor (EGFR) are in clinical use or late stages of clinical development. Here we investigate the molecular basis for inhibition of EGFR activation by the therapeutic antibody matuzumab (EMD72000). We describe the X-ray crystal structure of the Fab fragment of matuzumab (Fab72000) in complex with isolated domain III from the extracellular region of EGFR. Fab72000 interacts with an epitope on EGFR that is distinct from the ligand-binding region on domain III and from the cetuximab/Erbitux epitope. Matuzumab blocks ligand induced receptor activation indirectly by sterically preventing the domain rearrangement and local conformational changes that must occur for high-affinity ligand binding and receptor dimerization. PMID- 18394560 TI - Advanced therapies for movement disorders in neurotherapeutics. PMID- 18394561 TI - Current status of symptomatic medical therapy in Parkinson's disease. AB - Symptomatic medical therapies for Parkinson's disease (PD) have been disease modifying and have led to improvement in daily function, quality of life, and survival. For 40 years, these therapies have been primarily dopaminergic, and currently include the dopamine (DA) precursor levodopa (LD), DA agonists, catechol-O-methyltransferase (COMT) inhibitors, and monoamine oxidase (MAO) inhibitors. The roles of all these classes of agents have evolved, with significant changes occurring since the early 2000s. This article reviews the current literature for each of these classes of drugs, with a focus on efficacy and place in the therapeutic scheme. Levodopa is no longer considered to be toxic and, thus, its early use is not only appropriate but recommended. Ergot agonists are no longer in use, and new agents administered in patch form or subcutaneous injections have been approved. The COMT inhibitor tolcapone, with its significant efficacy, has been reintroduced, and two new MAO inhibitors have been approved. Selected safety issues are discussed, including the incidence of melanoma in relation to LD; pathological gambling and DA agonists; hepatic toxicity of tolcapone; and the tyramine or so-called cheese reaction with MAO B inhibitors. The article closes with a discussion of future directions and new drugs under development. PMID- 18394562 TI - Symptomatic treatment of Huntington disease. AB - Huntington disease (HD) is a progressive heredoneurodegenerative disease manifested by chorea and other hyperkinetic (dystonia, myoclonus, tics) and hypokinetic (parkinsonism) movement disorders. In addition, a variety of psychiatric and behavioral symptoms, along with cognitive decline, contribute significantly to the patient's disability. Because there are no effective neuroprotective therapies that delay the progression of the disease, symptomatic treatment remains the cornerstone of medical management. Several classes of medications have been used to ameliorate the various symptoms of HD, including typical and atypical neuroleptics, dopamine depleters, antidepressants, antiglutamatergic drugs, GABA agonists, antiepileptic medications, acetylcholinesterase inhibitors, and botulinum toxin. Recently, surgical approaches including pallidotomy, deep brain stimulation, and fetal cell transplants have been used for the symptomatic treatment of HD. The selected therapy must be customized to the needs of each patient, minimizing the potential adverse effects. The primary aim of this article is to review the role of the different therapies, both available and investigational, for the treatment of the motor, psychiatric, behavioral, and cognitive symptoms of HD, and to examine their impact on the patient's functionality and quality of life. PMID- 18394563 TI - Experimental therapeutics for dystonia. AB - Dystonia is a neurological syndrome characterized by excessive involuntary muscle contractions leading to twisting movements and unnatural postures. It has many different clinical manifestations, and many different causes. More than 3 million people worldwide suffer from dystonia, yet there are few broadly effective treatments. In the past decade, progress in research has advanced our understanding of the pathogenesis of dystonia to a point where drug discovery efforts are now feasible. Several strategies can be used to develop novel therapeutics for dystonia. Existing therapies have only modest efficacy, but may be refined and improved to increase benefits while reducing side effects. Identifying rational targets for drug intervention based on the pathogenesis of dystonia is another strategy. The surge in both basic and clinical research discoveries has provided insights at all levels, including etiological, physiological and nosological, to enable such a targeted approach. The empirical approach to drug discovery, whereby compounds are identified using a nonmechanistic strategy, is complementary to the rational approach. With the recent development of multiple animal models of dystonia, it is now possible to develop assays and perform drug screens on vast numbers of compounds. This multifaceted approach to drug discovery in dystonia will likely provide lead compounds that can then be translated for clinical use. PMID- 18394564 TI - Protection against Parkinson's disease progression: clinical experience. AB - Treatments with potential neuroprotective capability for Parkinson's disease (PD) have been investigated in randomized, controlled, clinical trials and other studies since the mid-1980s. Although promising leads have arisen, no therapy has been proven to halt or slow disease progression. Several large-scale studies have highlighted progress in methodology, as well as the frustrations of translating laboratory science to practical applications. This review summarizes findings from clinical trials with several classes of compounds, including monoamine oxidase-B inhibitors (selegiline, lazabemide, rasagiline), dopaminergic drugs (ropinirole, pramipexole, levodopa), antioxidant strategies (alpha-tocopherol), mitochondrial energy enhancers (coenzyme Q(10), creatine), antiapoptotic agents (TCH346, minocycline, CEP-1347), and antiglutamatergic compounds (riluzole). Beyond small-molecule pharmacology, gene therapy approaches, such as delivering neurotrophic substances (e.g., neurturin) by viral vector, are the next generation of treatment options. PMID- 18394565 TI - Neuroprotection for Huntington's disease: ready, set, slow. AB - The ultimate goal for Huntington's disease (HD) therapeutics is to develop disease-modifying neuroprotective therapies that can delay or prevent illness in those who are at genetic risk and can slow progression in those who are affected clinically. Neuroprotection is the preservation of neuronal structure, function, and viability, and neuroprotective therapy is thus targeted at the underlying pathology of HD, rather than at its specific symptoms. Preclinical target discovery research in HD is identifying numerous distinct targets, along with options for modulating them, with some proceeding into large-scale efficacy studies in early symptomatic HD subjects. The first pilot studies of neuroprotective compounds in premanifest HD are also soon to begin. This review discusses the opportunities for neuroprotection in HD, clinical methodology in premanifest and manifest HD, the clinical assessment of neuroprotection, molecular targets and therapeutic leads, and the current state of clinical development. PMID- 18394567 TI - Spheramine for treatment of Parkinson's disease. AB - Spheramine (Bayer Schering Pharma AG, Berlin, Germany) is currently being tested as a new approach for the treatment of Parkinson's disease (PD). It consists of an active component of cultured human retinal pigment epithelial (hRPE) cells, attached to an excipient part of cross-linked porcine gelatin microcarrriers. Spheramine is administered by stereotactic implantation into the striatum of PD patients and the use of immunosuppression is not required. Current pharmacologic therapies of PD are oriented to the administration of dopaminergic medications. Human RPE cells produce levodopa, and this constitutes the rationale to use Spheramine for the treatment of PD. The preclinical development of Spheramine included extensive biologic, pharmacologic, and toxicologic studies in vitro and in animal models of PD. The first clinical trial in humans evaluated the safety and efficacy of Spheramine implanted in the postcommissural putamen contralateral to the most affected side in six patients with advanced PD. This open-label study demonstrated good tolerability and showed sustained motor clinical improvement. A phase II double-blind, randomized, multicenter, placebo-controlled (sham surgery) study is underway to evaluate safety, tolerability, and efficacy of Spheramine implanted bilaterally into the postcommissural putamen of patients with advanced PD. Spheramine represents a treatment approach with the potential of supplying a more continuous delivery of levodopa to the striatum in advanced PD than can be achieved with oral therapy alone. PMID- 18394566 TI - Therapeutic potentials of human embryonic stem cells in Parkinson's disease. AB - The loss of dopaminergic neurons of the substantia nigra is the pathological hallmark characteristic of Parkinson's disease (PD). The strategy of replacing these degenerating neurons with other cells that produce dopamine has been the main approach in the cell transplantation field for PD research. The isolation, differentiation, and long-term cultivation of human embryonic stem cells and the therapeutic research discovery made in relation to the beneficial properties of neurotrophic and neural growth factors has advanced the transplantation field beyond dopamine-producing cells. The present review addresses recent advances in human embryonic stem cell experimentation in relation to treating PD, as well as cell transplantation techniques in conjunction with alternative therapeutics. PMID- 18394568 TI - Advances in gene therapy for movement disorders. AB - After nearly 20 years of preclinical experimentation with various gene delivery approaches in animal models of Parkinson's disease (PD), clinical trials are finally underway. The risk/benefit ratio for these procedures is now generally considered acceptable under approved protocols. The current vehicle for gene delivery to the human brain is recombinant adeno-associated viral vector, which is nonpathogenic and non-self-amplifying. Candidate genes tested in PD patients encode 1) glutamic acid decarboxylase, which is injected into the subthalamic nucleus to catalyze biosynthesis of the inhibitory neurotransmitter gamma aminobutyric acid and so essentially mimic deep brain stimulation of this nucleus; 2) aromatic l-amino acid decarboxylase, which converts l-dopa to dopamine; and 3) neurturin, a member of the glial cell line-derived neurotrophic factor family. Unraveling the genetic underpinnings of PD could allow gene therapy to go beyond modulating neurotransmission or providing trophic effects to dopaminergic neurons by delivering a specific missing or defective gene. For example, the parkin gene (PARK2) is linked to recessively inherited PD due to loss of function mutations; it prevents alpha-synuclein-induced degeneration of nigral dopaminergic neurons in rats and nonhuman primates. On the other hand, for dominantly inherited Huntington's disease (HD), in which an expanded polyglutamine tract imparts to the protein huntingtin a toxic gain of function, repressing expression of the mutant allele in the striatum using RNA interference technology mitigates pathology and delays the phenotype in a mouse model. Here we review the current state of preclinical and clinical gene therapy studies conducted in PD and HD. PMID- 18394569 TI - Treatment of Parkinson's disease with trophic factors. AB - Trophic factors are proteins that support and protect subpopulations of cells. A number have been reported to act on dopaminergic neurons in vitro and in vivo, making them potential therapeutic candidates for Parkinson's disease. All of these candidate factors protect dopaminergic neurons if given prior to, or with, selective neurotoxins. Fewer trophic factors, primarily glial-derived neurotrophic factor (GDNF) and its relative, neurturin (NRTN; also known as NTN), have been shown to restore function in damaged dopamine neurons after the acute effects of neurotoxins have subsided. A major barrier to clinical translation has been delivery. GDNF delivered by intracerebroventricular injection in patients was ineffective, probably because GDNF did not reach the target, the putamen, and intraputaminal infusion was ineffective, probably because of limited distribution within the putamen. A randomized clinical trial with gene therapy for NRTN is underway, in an attempt to overcome these problems with targeting and distribution. Other strategies are available to induce trophic effects in the CNS, but have not yet been the focus of human research. To date, clinical trials have focused on restoration of function (i.e., improvement of parkinsonism). Protection (i.e., slowing or halting disease progression and functional decline) might be a more robust effect of trophic agents. Laboratory research points to their effectiveness in protecting neurons and even restoring dopaminergic function after a monophasic neurotoxic insult. Utility for such compounds in patients with Parkinson's disease and ongoing loss of dopaminergic neurons remains to be proven. PMID- 18394570 TI - What happened to posteroventral pallidotomy for Parkinson's disease and dystonia? AB - Fifteen years after its resurrection, pallidotomy for Parkinson's disease (PD) and dystonia has once again been supplanted, this time by deep brain stimulation (DBS). Did this occur because pallidotomy was not effective or safe, or because DBS was found to be more effective and safer? This review focuses on the evidence and its quality-supporting the effectiveness and safety of pallidotomy for PD and dystonia, and the comparative effectiveness and safety of DBS of the subthalamic nucleus (STN) and globus pallidus pars interna (GPi). Discussed first are the determinants of "level 1" recommendations, including the confounding effects on interpretation of randomized clinical trials (RCTs) that fail to control for patient bias (i.e., placebo effects). Although several RCTs have been performed comparing unilateral pallidotomy to medical therapy, GPi DBS, or STN DBS for PD, none controlled for patient bias. Comparison of these trials to estimate the placebo effect, and examination of retrospective case series, suggests that the true effectiveness of unilateral pallidotomy is 20% to 30% reduction of 'off' total motor UPDRS scores, which is similar to the effects of unilateral GPi DBS or STN DBS, but less than bilateral STN DBS. At experienced centers, safety of unilateral pallidotomy appears equivalent to unilateral DBS, but bilateral DBS is likely safer than bilateral pallidotomy. Whereas there have been no RCTs of pallidotomy for dystonia, two double-blind, sham-controlled RCTs of bilateral GPi DBS were performed. Nevertheless, limited uncontrolled series suggest that bilateral pallidotomy is similar to GPi DBS in effectiveness and safety for dystonia. Thus, pallidotomy was not rejected because of lack of effectiveness or safety, and it remains a viable alternative in situations where DBS is not available or not feasible. PMID- 18394572 TI - Deep brain stimulation for Parkinson's disease. AB - The surgical treatment of Parkinson's disease has been through a revival phase over the last 20 years with the development of deep brain stimulation (DBS). Thalamic DBS was developed first and has proven to be a very effective treatment for tremor. The limitation is the lack of effect on other symptoms. Other targets were therefore investigated, and the procedure was applied to the subthalamic nucleus (STN) and the internal globus pallidus (GPi). STN stimulation can improve a wide range of symptoms and is currently the preferred target for many patients. Nevertheless, the morbidity seems higher than with other targets, and the selection criteria have to be quite strict. When STN DBS is not advised, thalamic DBS remains an option for patients with severe tremor, and GPi stimulation for those with severe dyskinesias. DBS remains a symptomatic treatment for a limited number of patients; it does not seem to alter the disease progression, and many patients are not suitable. There is, therefore, the need for further research into other targets and other approaches. PMID- 18394571 TI - Mechanisms and targets of deep brain stimulation in movement disorders. AB - Chronic electrical stimulation of the brain, known as deep brain stimulation (DBS), has become a preferred surgical treatment for medication-refractory movement disorders. Despite its remarkable clinical success, the therapeutic mechanisms of DBS are still not completely understood, limiting opportunities to improve treatment efficacy and simplify selection of stimulation parameters. This review addresses three questions essential to understanding the mechanisms of DBS. 1) How does DBS affect neuronal tissue in the vicinity of the active electrode or electrodes? 2) How do these changes translate into therapeutic benefit on motor symptoms? 3) How do these effects depend on the particular site of stimulation? Early hypotheses proposed that stimulation inhibited neuronal activity at the site of stimulation, mimicking the outcome of ablative surgeries. Recent studies have challenged that view, suggesting that although somatic activity near the DBS electrode may exhibit substantial inhibition or complex modulation patterns, the output from the stimulated nucleus follows the DBS pulse train by direct axonal excitation. The intrinsic activity is thus replaced by high-frequency activity that is time-locked to the stimulus and more regular in pattern. These changes in firing pattern are thought to prevent transmission of pathologic bursting and oscillatory activity, resulting in the reduction of disease symptoms through compensatory processing of sensorimotor information. Although promising, this theory does not entirely explain why DBS improves motor symptoms at different latencies. Understanding these processes on a physiological level will be critically important if we are to reach the full potential of this powerful tool. PMID- 18394574 TI - Deep brain stimulation and tremor. AB - Deep brain stimulation (DBS) has been used to treat various tremor disorders for several decades. Medication-resistant, disabling essential tremor (ET) is the most common tremor disorder treated with DBS. The treatment has been consistently reported to result in significant benefit in upper extremity, as well as head and voice tremor, all of which were improved more dramatically with bilateral procedures. These benefits have been demonstrated to be sustained for up to 7 years. DBS has also been shown to be beneficial for the tremor associated with multiple sclerosis and post-traumatic tremor; however, fewer cases have been reported and the benefit is less consistent, less dramatic, and more transient than that seen with ET. The ventral intermediate nucleus of the thalamus is the most common DBS target for tremor disorders, but more recent studies have demonstrated benefits in tremor from DBS of the subthalamic area, primarily the zona incerta. Surgical complications are relatively uncommon and are generally less frequent than those seen with thalamotomy. Stimulation-related effects are usually mild and resolve with adjustment of stimulation parameters. DBS is thus a relatively safe and effective treatment for tremor disorders, particularly for medication-resistant, disabling ET, but may also have some role in medication resistant, disabling tremor associated with multiple sclerosis and traumatic head injury. PMID- 18394573 TI - Treatment of dystonia with deep brain stimulation. AB - Pallidal deep brain stimulation (DBS) is an established treatment option for medically refractive dystonia. The mechanism by which globus pallidus pars interna (GPi) DBS improves dystonia is still unclear. Primary generalized dystonia usually responds well to this therapy, as recently confirmed in two well designed, double-blind, controlled trials; however, predictors of outcome within this population are not well known. The role of GPi DBS in idiopathic cervical dystonia resistant to treatment with botulinum toxin, in tardive dystonia, and in some types of secondary dystonia are emerging as populations of patients who may also benefit, but outcomes are not well documented. Serious complications from this therapy are rare. Future research will likely continue to address the most appropriate programming settings for various populations of dystonia, the mechanism by which DBS affects dystonia, and the possibility of alternative brain targets that might have less associated side effects or greater efficacy than the GPi. PMID- 18394575 TI - Deep brain stimulation in Tourette's Syndrome. AB - Tourette's Syndrome (TS) is a neuropsychiatric disorder characterized by motor and vocal tics, often associated with behavioral disorders. Symptoms often disappear before or during adulthood. The pathophysiology of TS is still a matter of considerable debate. Current knowledge of cortico-basal ganglia thalamocortical circuits provide explanations for the beneficial effects of deep brain stimulation (DBS) on tics. When conservative treatment fails in patients with severe TS, DBS may be a therapeutic option. In 1999, thalamic DBS was introduced for intractable TS. Since then, multiple targets have been used in a small number of patients, including the globus pallidus pars interna and the nucleus accumbens. Inclusion and exclusion criteria have been formulated to identify good candidates for DBS. PMID- 18394577 TI - Characterization of ion-exchange membrane materials: properties vs structure. AB - This review focuses on the preparation, structure and applications of ion exchange membranes formed from various materials and exhibiting various functions (electrodialytic, perfluorinated sulphocation-exchange and novel laboratory tested membranes). A number of experimental techniques for measuring electrotransport properties as well as the general procedure for membrane testing are also described. The review emphasizes the relationships between membrane structures, physical and chemical properties and mechanisms of electrochemical processes that occur in charged membrane materials. The water content in membranes is considered to be a key factor in the ion and water transfer and in polarization processes in electromembrane systems. We suggest the theoretical approach, which makes it possible to model and characterize the electrochemical properties of heterogeneous membranes using several transport-structural parameters. These parameters are extracted from the experimental dependences of specific electroconductivity and diffusion permeability on concentration. The review covers the most significant experimental and theoretical research on ion exchange membranes that have been carried out in the Membrane Materials Laboratory of the Kuban State University. These results have been discussed at the conferences "Membrane Electrochemistry", Krasnodar, Russia for many years and were published mainly in Russian scientific sources. PMID- 18394576 TI - Noninvasive brain stimulation for Parkinson's disease and dystonia. AB - Repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS) are promising noninvasive cortical stimulation methods for adjunctive treatment of movement disorders. They avoid surgical risks and provide theoretical advantages of specific neural circuit neuromodulation. Neuromodulatory effects depend on extrinsic stimulation factors (cortical target, frequency, intensity, duration, number of sessions), intrinsic patient factors (disease process, individual variability and symptoms, state of medication treatment), and outcome measures. Most studies to date have shown beneficial effects of rTMS or tDCS on clinical symptoms in Parkinson's disease (PD) and support the notion of spatial specificity to the effects on motor and nonmotor symptoms. Stimulation parameters have varied widely, however, and some studies are poorly controlled. Studies of rTMS or tDCS in dystonia have provided abundant data on physiology, but few on clinical effects. Multiple mechanisms likely contribute to the clinical effects of rTMS and tDCS in movement disorders, including normalization of cortical excitability, rebalancing of distributed neural network activity, and induction of dopamine release. It remains unclear how to individually adjust rTMS or tDCS factors for the most beneficial effects on symptoms of PD or dystonia. Nonetheless, the noninvasive nature, minimal side effects, positive effects in preliminary clinical studies, and increasing evidence for rational mechanisms make rTMS and tDCS attractive for ongoing investigation. PMID- 18394578 TI - Identification of the SPG15 gene, encoding spastizin, as a frequent cause of complicated autosomal-recessive spastic paraplegia, including Kjellin syndrome. AB - Hereditary spastic paraplegias (HSPs) are genetically and phenotypically heterogeneous disorders. Both "uncomplicated" and "complicated" forms have been described with various modes of inheritance. Sixteen loci for autosomal-recessive "complicated" HSP have been mapped. The SPG15 locus was first reported to account for a rare form of spastic paraplegia variably associated with mental impairment, pigmented maculopathy, dysarthria, cerebellar signs, and distal amyotrophy, sometimes designated as Kjellin syndrome. Here, we report the refinement of SPG15 to a 2.64 Mb genetic interval on chromosome 14q23.3-q24.2 and the identification of ZFYVE26, which encodes a zinc-finger protein with a FYVE domain that we named spastizin, as the cause of SPG15. Six different truncating mutations were found to segregate with the disease in eight families with a phenotype that included variable clinical features of Kjellin syndrome. ZFYVE26 mRNA was widely distributed in human tissues, as well as in rat embryos, suggesting a possible role of this gene during embryonic development. In the adult rodent brain, its expression profile closely resembled that of SPG11, another gene responsible for complicated HSP. In cultured cells, spastizin colocalized partially with markers of endoplasmic reticulum and endosomes, suggesting a role in intracellular trafficking. PMID- 18394579 TI - A mutation in HOXA2 is responsible for autosomal-recessive microtia in an Iranian family. AB - Microtia, a congenital deformity manifesting as an abnormally shaped or absent external ear, occurs in one out of 8,000-10,000 births. We ascertained a consanguineous Iranian family segregating with autosomal-recessive bilateral microtia, mixed symmetrical severe to profound hearing impairment, and partial cleft palate. Genome-wide linkage analysis localized the responsible gene to chromosome 7p14.3-p15.3 with a maximum multi-point LOD score of 4.17. In this region, homeobox genes from the HOXA cluster were the most interesting candidates. Subsequent DNA sequence analysis of the HOXA1 and HOXA2 homeobox genes from the candidate region identified an interesting HOXA2 homeodomain variant: a change in a highly conserved amino acid (p.Q186K). The variant was not found in 231 Iranian and 109 Belgian control samples. The critical contribution of HoxA2 for auditory-system development has already been shown in mouse models. We built a homology model to predict the effect of this mutation on the structure and DNA-binding activity of the homeodomain by using the program Modeler 8v2. In the model of the mutant homeodomain, the position of the mutant lysine side chain is consistently farther away from a nearby phosphate group; this altered position results in the loss of a hydrogen bond and affects the DNA-binding activity. PMID- 18394580 TI - Mapping of small RNAs in the human ENCODE regions. AB - The elucidation of the largely unknown transcriptome of small RNAs is crucial for the understanding of genome and cellular function. We report here the results of the analysis of small RNAs (< 50 nt) in the ENCODE regions of the human genome. Size-fractionated RNAs from four different cell lines (HepG2, HelaS3, GM06990, SK N-SH) were mapped with the forward and reverse ENCODE high-density resolution tiling arrays. The top 1% of hybridization signals are termed SmRfrags (Small RNA fragments). Eight percent of SmRfrags overlap the GENCODE genes (CDS), given that the majority map to intergenic regions (34%), intronic regions (53%), and untranslated regions (UTRs) (5%). In addition, 9.6% and 16.8% of SmRfrags in the 5' UTR regions overlap significantly with His/Pol II/TAF250 binding sites and DNase I Hypersensitive sites, respectively (compared to the 5.3% and 9% expected). Interestingly, 17%-24% (depending on the cell line) of SmRfrags are sense-antisense strand pairs that show evidence of overlapping transcription. Only 3.4% and 7.2% of SmRfrags in intergenic regions overlap transcribed fragments (Txfrags) in HeLa and GM06990 cell lines, respectively. We hypothesized that a fraction of the identified SmRfrags corresponded to microRNAs. We tested by Northern blot a set of 15 high-likelihood predictions of microRNA candidates that overlap with smRfrags and validated three potential microRNAs ( approximately 20 nt length). Notably, most of the remaining candidates showed a larger hybridizing band ( approximately 100 nt) that could be a microRNA precursor. The small RNA transcriptome is emerging as an important and abundant component of the genome function. PMID- 18394582 TI - Predictors of re-excision for positive or close margins in breast-conservation therapy for pT1 tumors. AB - BACKGROUND: An increasing number of patients with early-stage breast cancer is being diagnosed by way of population-wide mammographic screening in women. Because breast-conservation therapy (BCT) is the standard treatment, the aim of our study was to determine factors predisposing patients to re-excision for pT1 tumors. METHODS: A retrospective study was conducted at Besancon University Hospital in France. Patients with pathologic diagnosis of invasive breast carcinoma <20 mm in size (pT1) and undergoing BCT were selected. From these, "no re-excision" and "re-excision" subgroups were constituted and compared in terms of patient and tumor characteristics. The intent of all surgeries was therapeutic. RESULTS: Of 206 patients who underwent BCT for pT1 tumors, 84 (41%) needed re-excision. After multivariate analysis, factors predicting re-excision were the absence of positive histologic diagnosis before surgery (P <.0001), limited superficial and deep surgical resection (P <.0001), multifocal lesion (P <.0001), and size of the in situ carcinoma (P <.0001). CONCLUSIONS: These predictive factors could be useful in reducing the rate of re-excision as well as in identifying patients with multifocal tumors and/or extensive in situ carcinoma whose disease would be better managed by mastectomy. PMID- 18394581 TI - Bayesian meta-analysis of genetic association studies with different sets of markers. AB - Robust assessment of genetic effects on quantitative traits or complex-disease risk requires synthesis of evidence from multiple studies. Frequently, studies have genotyped partially overlapping sets of SNPs within a gene or region of interest, hampering attempts to combine all the available data. By using the example of C-reactive protein (CRP) as a quantitative trait, we show how linkage disequilibrium in and around its gene facilitates use of Bayesian hierarchical models to integrate informative data from all available genetic association studies of this trait, irrespective of the SNP typed. A variable selection scheme, followed by contextualization of SNPs exhibiting independent associations within the haplotype structure of the gene, enhanced our ability to infer likely causal variants in this region with population-scale data. This strategy, based on data from a literature based systematic review and substantial new genotyping, facilitated the most comprehensive evaluation to date of the role of variants governing CRP levels, providing important information on the minimal subset of SNPs necessary for comprehensive evaluation of the likely causal relevance of elevated CRP levels for coronary-heart-disease risk by Mendelian randomization. The same method could be applied to evidence synthesis of other quantitative traits, whenever the typed SNPs vary among studies, and to assist fine mapping of causal variants. PMID- 18394583 TI - [Statin in prevention of periprocedural myonecrosis after percutaneous coronary angioplasty: systematic review and meta-analysis]. AB - Postprocedural myocardial necrosis as indicated by biomarkers increase is frequent after percutaneous coronary interventions (PCI) in up to 40% of cases. Pretreatment by statin could reduce the occurrence of those periprocedural myonecrosis. Medline and Cochrane Database screenings allow us to select articles published until March 2007. Sixteen prospective and retrospective studies have been identified in relation with the impact of statin pretreatment on periprocedural myonecrosis or morbimortality outcomes in patients who underwent PCI. Among selected studies, 12 provide the rate of periprocedural myocardial infarction. The relative risk (RR) of periprocedural myocardial necrosis in patients pretreated with statin was 0.50 (95% CI: 0.40-0.63). Among the 5409 patients with clinical follow-up ensured until six months, the RR of death in patients pretreated with statin before the PCI was 0.64 (95% CI: 0.44-0.93). Significant heterogeneity was observed in the results. In sensitivity analysis, stable and unstable patients were compared with similar RR reduction of periprocedural myonecrosis in favour of patients receiving statin before PCI. In the present meta-analysis, we confirm that statin pretreatment before PCI is able to reduce periprocedural myonecrosis and suggest a potential impact on six months survival of patients. PMID- 18394584 TI - [Evaluation of the benefit of early valve surgery in the treatment of infective endocarditis. Contribution of propensity analyses]. AB - Early valve surgery for infective endocarditis, that is surgery performed during the course of antibiotic therapy, is widely used. However, this intervention has never been proven effective in any clinical trial. Five recently published observational studies, using propensity analysis, analyzed the relationship between surgery and mortality. But they gave conflicting results, mainly because of their methodological limitations. Thus, to date, there is no reliable estimation of early valvular surgery impact on mortality. In addition, this example allows for discussing the interest of propensity analyses and to emphasizing their limitations. PMID- 18394585 TI - [Use of prolonged-release nicotinic acid in patients treated with statins as a secondary prevention and a persistently low HDL-cholesterol level in France]. AB - HDL-c is a potent predictor of cardiovascular risk in all epidemiologic studies, especially as secondary prevention, regardless of LDL-c level obtained on statin treatment. The objective of this longitudinal observational study was to investigate the effects of prolonged-release nicotinic acid in high cardiovascular risk patients on statin treatment and with low HDL-c. METHOD: The selected patients of both genders, aged over 18 years patients showed primary hypercholesterolemia or mixed hyperlipidemia, received statin treatment as secondary prevention and had a HDL-c level 3)-alpha-D GalAp-(1-->4)-alpha-D-GalAp-(1-->4)-alpha-D-GalAp-(1-->4)-GalAp. The most abundant was beta-D-Xylp-(1-->3)-alpha-D-GalAp-(1-->4)-alpha-D-GalAp-(1-->4)(beta D-Xylp-(1-->3)-alpha-D-GalAp-(1-->4))-alpha-D-GalAp-(1-->4)-alpha-D-GalAp-(1-->4) GalAp. Given that the nonreducing ends of the oligosaccharides often were xylosylated GalA residues, and that fungal EPG digests homogalacturonans between the third and fourth GalA bound to the enzyme, it appears that EPG can accommodate a xylosylated GalA in the site that binds the fourth GalA. Since all of the oligosaccharides characterized had three unsubstituted GalA residues at their reducing ends, the enzyme appears not to accommodate xylosylated residues in the first three sugar-binding sites. Thus, XGA regions with fewer than three unsubstituted residues between branch points will be resistant to EPG. The EPG susceptible XGA was not recovered from cell walls prepared using phosphate buffer for the homogenization of the watermelon tissue, probably because it was degraded by endogenous watermelon EPG and lost during isolation of the walls. Use of Tris buffered phenol during wall isolation to prevent enzyme action caused some amidation of GalA residues with Tris. PMID- 18394595 TI - Cancer classification using Rotation Forest. AB - We address the microarray dataset based cancer classification using a newly proposed multiple classifier system (MCS), referred to as Rotation Forest. To the best of our knowledge, it is the first time that Rotation Forest has been applied to the microarray dataset classification. In the framework of Rotation Forest, a linear transformation method is required to project data into new feature space for each classifier, and then the base classifiers are trained in different new spaces so as to enhance both the accuracies of base classifiers and the diversity in the ensemble system. Principal component analysis (PCA), non-parametric discriminant analysis (NDA) and random projections (RP) were applied to feature transformation in the original Rotation Forest. In this paper, we use independent component analysis (ICA) as a new transformation method since it can better describe the property of microarray data. The breast cancer dataset and prostate dataset are deployed to validate the efficiency of Rotation Forest. In all the experiments, it can be found that Rotation Forest outperforms other MCSs, such as Bagging and Boosting. In addition, the experimental results also revealed that ICA can further improve the performance of Rotation Forest compared with the original transformation methods. PMID- 18394597 TI - Prepatterning the Drosophila notum: the three genes of the iroquois complex play intrinsically distinct roles. AB - The Drosophila thorax exhibits 11 pairs of large sensory organs (macrochaetes) identified by their unique position. Remarkably precise, this pattern provides an excellent model system to study the genetic basis of pattern formation. In imaginal wing discs, the achaete-scute proneural genes are expressed in clusters of cells that prefigure the positions of each macrochaete. The activities of prepatterning genes provide positional cues controlling this expression pattern. The three homeobox genes clustered in the iroquois complex (araucan, caupolican and mirror) are such prepattern genes. mirror is generally characterized as performing functions predominantly different from the other iroquois genes. Conversely, araucan and caupolican are described in previous studies as performing redundant functions in most if not all processes in which they are involved. We have addressed the question of the specific role of each iroquois gene in the prepattern of the notum and we clearly demonstrate that they are intrinsically different in their contribution to this process: caupolican and mirror, but not araucan, are required for the neural patterning of the lateral notum. However, when caupolican and/or mirror expression is reduced, araucan loss of function has an effect on thoracic bristles development. Moreover, the overexpression of araucan is able to rescue caupolican loss of function. We conclude that, although retaining some common functionalities, the Drosophila iroquois genes are in the process of diversification. In addition, caupolican and mirror are required for stripe expression and, therefore, to specify the muscular attachment sites prepattern. Thus, caupolican and mirror may act as common prepattern genes for all structures in the lateral notum. PMID- 18394596 TI - Multiple roles for Med12 in vertebrate endoderm development. AB - In zebrafish, the endoderm originates at the blastula stage from the most marginal blastomeres. Through a series of complex morphogenetic movements and differentiation events, the endodermal germ layer gives rise to the epithelial lining of the digestive tract as well as its associated organs such as the liver, pancreas, and swim bladder. How endodermal cells differentiate into distinct cell types such as hepatocytes or endocrine and exocrine pancreatic cells remains a major question. In a forward genetic screen for genes regulating endodermal organ development, we identified mutations at the shiri locus that cause defects in the development of a number of endodermal organs including the liver and pancreas. Detailed phenotypic analyses indicate that these defects are partially due to a reduction in endodermal expression of the hairy/enhancer of split-related gene, her5, at mid to late gastrulation stages. Using the Tg(0.7her5:EGFP)(ne2067) line, we show that her5 is expressed in the endodermal precursors that populate the pharyngeal region as well as the organ-forming region. We also find that knocking down her5 recapitulates some of the endodermal phenotypes of shiri mutants, further revealing the role of her5 in endoderm development. Positional cloning reveals that shiri encodes Med12, a regulatory subunit of the transcriptional Mediator complex recently associated with two human syndromes. Additional studies indicate that Med12 modulates the ability of Casanova/Sox32 to induce sox17 expression. Thus, detailed phenotypic analyses of embryos defective in a component of the Mediator complex have revealed new insights into discrete aspects of vertebrate endoderm development, and provide possible explanations for the craniofacial and digestive system defects observed in humans with mutations in MED12. PMID- 18394598 TI - Emerging novel functions of RNAs, and binary phenotype? AB - Loss-of-function technology has been one of the most popular knockout tools for the study of gene function in cell and developmental biology. This technology employs two basic approaches for elimination of the protein of interest. The morpholino antisense oligonucleotides approach relies on inhibiting translation of the given protein without degrading the cognate mRNA. The antisense deoxynucleotides and siRNA approach acts via removal of the mRNA template, which then prevents protein translation. In the latest approach, as well as in these genetic knockout approaches that eliminate or alter the level of mRNA transcribed from the gene of interest, the assumption is and always has been that the only relevant function of mRNA is to make a protein, and, thus, the effect of removing mRNA equals the effect of removing its protein function. However, the most recent studies of different biological systems point to completely novel and unexpected functions of the subpopulation of localized RNAs and suggest that, at least in some cases, the normal cell or embryo phenotype is in fact binary i.e. depends not only on the function of the protein but also on the autonomous function of its mRNA. PMID- 18394600 TI - Epigenetic and HIF-1 regulation of stanniocalcin-2 expression in human cancer cells. AB - Mammalian stanniocalcin-2 (STC2) is a secreted glycoprotein hormone with a putative role in unfolded protein response and apoptosis. Here we reported that STC2 expression was sporadically abrogated in human cancer cells by transcriptional silencing associated with CpG island promoter hypermethylation. Direct sequencing of bisulfite-modified DNA from a panel of seven human cancer cell lines revealed that CpG dinucleotides in STC2 promoter was methylated in human ovarian epithelial cancer (SKOV3, OVCAR3 and CaOV3), pancreatic cancer (BxP3), colon adenoma (HT29), and leukemia (Jurkat cells). STC2 CpG island hypermethylation was accompanied with a low basal STC2 expression level. Treatment of these cancer cells with 5-aza-2'-deoxycytidine (5-aza-CdR), an inhibitor of DNA methylation significantly induced STC2 expression. Using SKOV3 cells as a model, the link between DNA demethylation and STC2 expression was consistently demonstrated with hydralazine treatment, which was shown to reduce the protein level of DNA methyltransferase 1 (DNMT1) but stimulated STC2 expression. Two human normal surface ovarian cell-lines (i.e. IOSE 29 and 398) showed no methylation at CpG dinucleotides in the examined promoter region and were accompanied with high basal STC2 levels. Hypoxia stimulated STC2 expression in SKOV3 cells was markedly increased in 5-aza-CdR pretreated cells, showing that DNA methylation may hinder the HIF-1 mediated activation. To elucidate this possibility, RNA interference studies confirmed that endogenous HIF-1 alpha was a key factor for STC2 gene activation as well as in the synergistic induction of STC2 expression in 5-aza-CdR pretreated cells. Chromatin immunoprecipitation (ChIP) assay demonstrated the binding of HIF-1 alpha to STC2 promoter. The binding was increased in 5-aza-CdR pretreated cells. Collectively, this is the first report to show that STC2 was aberrantly hypermethylated in human cancer cells. The findings demonstrated that STC2 epigenetic inactivation may interfere with HIF-1 mediated activation of STC2 expression. PMID- 18394601 TI - Identification of a novel centrosomal protein CrpF46 involved in cell cycle progression and mitosis. AB - A novel centrosome-related protein CrpF46 was detected using a serum F46 from a patient suffering from progressive systemic sclerosis. We identified the protein by immunoprecipitation and Western blotting followed by tandem mass spectrometry sequencing. The protein CrpF46 has an apparent molecular mass of ~60 kDa, is highly homologous to a 527 amino acid sequence of the C-terminal portion of the protein Golgin-245, and appears to be a splice variant of Golgin-245. Immunofluorescence microscopy of synchronized HeLa cells labeled with an anti CrpF46 monoclonal antibody revealed that CrpF46 localized exclusively to the centrosome during interphase, although it dispersed throughout the cytoplasm at the onset of mitosis. Domain analysis using CrpF46 fragments in GFP-expression vectors transformed into HeLa cells revealed that centrosomal targeting is conferred by a C-terminal coiled-coil domain. Antisense CrpF46 knockdown inhibited cell growth and proliferation and the cell cycle typically stalled at S phase. The knockdown also resulted in the formation of poly-centrosomal and multinucleate cells, which finally became apoptotic. These results suggest that CrpF46 is a novel centrosome-related protein that associates with the centrosome in a cell cycle-dependent manner and is involved in the progression of the cell cycle and M phase mechanism. PMID- 18394599 TI - Myt1 and Ngn3 form a feed-forward expression loop to promote endocrine islet cell differentiation. AB - High levels of Ngn3 expression in pancreatic progenitor cells are both necessary and sufficient to initiate endocrine differentiation. While it is clear that the Notch-Hes1-mediated signals control the number of Ngn3-expressing cells in the developing pancreas, it is not known what factors control the level of Ngn3 expression in individual pancreatic cells. Here we report that Myt1b and Ngn3 form a feed-forward expression loop that regulates endocrine differentiation. Myt1b induces glucagon expression by potentiating Ngn3 transcription in pancreatic progenitors. Vice versa, Ngn3 protein production induces the expression of Myt1. Furthermore, pancreatic Myt1 expression largely, but not totally, relies on Ngn3 activity. Surprisingly, a portion of Myt1 expressing pancreatic cells express glucagon and other alpha cell markers in Ngn3 nullizygous mutant animals. These results demonstrate that Myt1b and Ngn3 positively regulate each other's expression to promote endocrine differentiation. In addition, the data uncover an unexpected Ngn3 expression-independent endocrine cell production pathway, which further bolsters the notion that the seemingly equivalent endocrine cells of each type, as judged by hormone and transcription factor expression, are heterogeneous in their origin. PMID- 18394602 TI - S-nitrosylation of proteins at the leading edge of migrating trophoblasts by inducible nitric oxide synthase promotes trophoblast invasion. AB - Nitric oxide regulates many important cellular processes including motility and invasion. Many of its effects are mediated through the modification of specific cysteine residues in target proteins, a process called S-nitrosylation. Here we show that S-nitrosylation of proteins occurs at the leading edge of migrating trophoblasts and can be attributed to the specific enrichment of inducible nitric oxide synthase (iNOS/NOS2) in this region. Localisation of iNOS to the leading edge is co-incidental with a site of extensive actin polymerisation and is only observed in actively migrating cells. In contrast endothelial nitric oxide synthase (eNOS/NOS3) shows distribution that is distinct and non-colocalised with iNOS, suggesting that the protein S-nitrosylation observed at the leading edge is caused only by iNOS and not eNOS. We have identified MMP-9 as a potential target for S-nitrosylation in these cells and demonstrate that it co-localises with iNOS at the leading edge of migrating cells. We further demonstrate that iNOS plays an important role in promoting trophoblast invasion, which is an essential process in the establishment of a successful pregnancy. PMID- 18394603 TI - Retinal ganglion cell neuroprotection in a rat model of glaucoma following brimonidine, latanoprost or combined treatments. AB - The aim of the present study is to evaluate the neuroprotective effect of two antiglaucomatous substances, regardless of their hypotensive effect in the eye. Brimonidine, which does not reduce IOP when administered intraperitoneally, and latanoprost, which has a renowned hypotensive effect topically. We examined rat retinal ganglion cell (RGC) survival and size distribution in experimental glaucoma in response to different glaucomatous agents. IOP was elevated by episcleral vein cauterization (EVC) prior to the application of different treatments: (I) PBS application (control group), (II) intraperitoneal administration of brimonidine (a general hypotensive agent), (III) topical application of latanoprost (an ocular hypotensive agent), and (IV) latanoprost combined with brimonidine. After 12 weeks, RGCs were retrogradely labeled with fluorogold and RGC density was analyzed. EVC caused a significant increase (42%) in IOP in each group before drug treatment. After 12weeks of EVC, RGC survival in control vs. EVC rats was 78.9+/-3.2%. No IOP reduction was observed in brimonidine injected rats, but RGC survival at 12 weeks was total (103.7+/-2.7%). In latanoprost treated rats, IOP dropped by around 22% and 94.7+/-3.7% of the RGC population survived. Finally in the latanoprost+brimonidine combined group, IOP was significantly reduced by 25% and 94.4+/-2.2% of RGCs survived. Surprisingly, whereas EVC led to a 6% increase in RGC soma size, brimonidine treatment was associated with a 9% reduction in the soma size of RGCs at 12 weeks. We conclude that brimonidine exerts a neuroprotective effect via a mechanism which is independent of IOP reduction. These findings indicate that cell survival in glaucoma may be enhanced by neuroprotective strategies which are independent of IOP reduction. No synergistic neuroprotective effect was observed when both treatments were applied simultaneously. PMID- 18394604 TI - Thalamic deep brain stimulation improves eyeblink conditioning deficits in essential tremor. AB - Several lines of evidence point to a disturbance of olivo-cerebellar pathways in essential tremor (ET). For example, subjects with ET exhibit deficits in eyeblink conditioning, a form of associative learning which is known to depend on the integrity of olivo-cerebellar circuits. Deep brain stimulation (DBS) of the ventrolateral thalamus is an established therapy for ET. If tremor in ET is related to the same pathology of the olivo-cerebellar system as impaired eyeblink conditioning, one may expect modulation of eyeblink conditioning by DBS. Delay eyeblink conditioning was assessed in 11 ET subjects treated with DBS (ET-DBS subjects) who were studied on two consecutive days with DBS switched off (day 1) and on (day 2). For comparison, 11 age-matched ET subjects without DBS (ET subjects) and 11 age-matched healthy controls were studied. On day 1, eyeblink conditioning was diminished in ET-DBS subjects and in ET subjects compared with controls. When DBS was switched on ET-DBS subjects exhibited conditioning rates within the range of controls on day 2, while ET subjects improved only minimally. Improved eyeblink conditioning in ET-DBS subjects suggests that thalamic DBS counteracts a functional disturbance of olivo-cerebellar circuits which is thought to be responsible for eyeblink conditioning deficits in ET. Modulation of cerebello-thalamic and/or thalamo-cortico-cerebellar pathways by DBS may play a role. PMID- 18394605 TI - Human neural stem cells migrate along the nigrostriatal pathway in a primate model of Parkinson's disease. AB - Although evidence of damage-directed neural stem cell (NSC) migration has been well-documented in the rodent, to our knowledge it has never been confirmed or quantified using human NSC (hNSC) in an adult non-human primate modeling a human neurodegenerative disease state. In this report, we attempt to provide that confirmation, potentially advancing basic stem cell concepts toward clinical relevance. hNSCs were implanted into the caudate nucleus (bilaterally) and substantia nigra (unilaterally) of 7, adult St. Kitts African green monkeys (Chlorocebus sabaeus) with previous exposure to systemic 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP), a neurotoxin that disrupts the dopaminergic nigrostriatal pathway. A detailed quantitative analysis of hNSC migration patterns at two time points (4 and 7 months) following transplantation was performed. Density contour mapping of hNSCs along the dorsal-ventral and medial lateral axes of the brain suggested that >80% of hNSCs migrated from the point of implantation to and along the impaired nigrostriatal pathway. Although 2/3 of hNSCs were transplanted within the caudate, <1% of 3x10(6) total injected donor cells were identified at this site. The migrating hNSC did not appear to be pursuing a neuronal lineage. In the striatum and nigrostriatal pathway, but not in the substantia nigra, some hNSCs were found to have taken a glial lineage. The property of neural stem cells to align themselves along a neural pathway rendered dysfunctional by a given disease is potentially a valuable clinical tool. PMID- 18394607 TI - Pregnancy outcome after preimplantation genetic diagnosis in an affected couple with X-linked adrenoleukodystrophy. AB - OBJECTIVE: To achieve a pregnancy free of X-linked adrenoleukodystrophy (X-ALD) by intracytoplasmic sperm injection (ICSI) and preimplantation genetic diagnosis (PGD). DESIGN: Case report. SETTING: Clinica FIV Recoletos, a private IVF center. PATIENT(S): A couple in which the man had X-ALD. INTERVENTION(S): The ICSI protocol and PGD of the obtained embryos. MAIN OUTCOME MEASURE(S): Blastomeres were analyzed by fluorescence in situ hybridization using sex selection techniques. Embryos were transferred and pregnancy was diagnosed by hCG analysis and ultrasonographic examination. RESULT(S): Ten embryos were obtained by ICSI. A biopsy was taken from eight embryos to perform PGD and two male embryos were transferred resulting in a twin pregnancy. CONCLUSION(S): This is the first registered gestation in which PGD has been used to prevent X-ALD transmission. PMID- 18394606 TI - Liposomal glucocorticosteroids in treatment of chronic autoimmune demyelination: long-term protective effects and enhanced efficacy of methylprednisolone formulations. AB - Liposomal encapsulation leads to enhanced efficacy of glucocorticosteroids (GS) in treatment of autoimmune diseases. Here we compare liposomal prednisolone (PL) to liposomal methylprednisolone (MPL) in chronic-relapsing myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis (EAE), a model closely reflecting aspects of multiple sclerosis (MS). At the maximum of the first relapse, a single dose of PL or MPL was applied at 10 mg/kg or at 4 mg/kg and compared to classical methylprednisolone (MP) pulse therapy. PL at 10 mg/kg was superior to free MP with long-term efficacy and a sustained protection even during the second and third relapse. At the same time, in vivo magnetic resonance imaging of rat brains revealed a significant reduction of T2-lesions after PL application. Comparison of PL and MPL at 10 mg/kg disclosed superior effects for MPL with an enhanced reduction of inflammatory infiltration as well as preservation of myelin and axons. Dose titration experiments underscored a dose dependent efficacy of liposomal GS with a sustained efficacy especially of the higher dosage. In histological analyses, PL10 was superior in reducing macrophage and T cell infiltration as well as demyelination and axonal loss while the lower dosages were still at least as effective as free MP. FACS analyses revealed an effect of liposome formulations on T cell numbers, the CD4/CD8 ratio, frequencies of regulatory T cells and adhesion molecule expression. In summary, liposomal GS and especially methylprednisolone formulations display an enhanced efficacy not only in acute inflammatory, but also in chronic demyelinating models of MS and confer long-term protection from relapses. These findings lay the groundwork for applying liposomal GS in clinical MS trials in the near future. PMID- 18394608 TI - Anejaculation as the presenting feature of pituitary microadenoma. AB - OBJECTIVE: To describe the resolution of anejaculation associated with PRL excess by dopamine agonist treatment. DESIGN: Case report. SETTING: Fertility clinic. PATIENT(S): Two men with anejaculation, in one case with associated infertility. INTERVENTION(S): Dopamine agonist treatment. MAIN OUTCOME MEASURE(S): Resolution of anejaculation and infertility. RESULT(S): Endocrine investigations in both men revealed low serum T, elevated PRL, and a pituitary microadenoma. Treatment with a dopamine agonist normalized serum hormone profiles and restored sexual function, with subsequent involution of the adenoma. CONCLUSION(S): Prolactinoma manifesting as anejaculation is rare but responds readily to treatment with a dopamine agonist. Serum PRL should be measured in any man presenting with anejaculation. PMID- 18394609 TI - Don't judge a book by its cover; a quintuplet pregnancy following transfer of five poor-quality embryos. AB - OBJECTIVE: To report a case of a quintuplet pregnancy following transfer of five poor-quality cellular-stage embryos. DESIGN: Case report. SETTING: University of Toronto-affiliated infertility clinic. PATIENT(S): A 35-year-old female with a 2 year history of secondary infertility. INTERVENTION(S): Intracytoplasmic sperm injection and embryo transfer. MAIN OUTCOME MEASURE(S): Clinical pregnancy. RESULT(S): Quintuplet implantation with five fetal hearts. CONCLUSION(S): Because the transfer of five poor embryos resulted in a quintuplet implantation, we recommend that embryos with significant morphologic abnormalities, especially in younger patients, be considered as viable embryos with implantation potential. PMID- 18394610 TI - A retroprospective study comparing three different assisted hatching techniques. AB - The purpose of this study was to determine the effect of four different assisted hatching techniques on pregnancy rates in women with prior IVF failure in fresh IVF cycles. The results suggested that assisted hatching utilizing laser, chemical, or microsurgical techniques increases both implantation and pregnancy rates. PMID- 18394611 TI - Laparoscopic cornuotomy using a temporary tourniquet suture and diluted vasopressin injection in interstitial pregnancy. AB - OBJECTIVE: To evaluate the efficiency of laparoscopic cornuotomy. DESIGN: Retrospective case review. SETTING: An urban medical center. PATIENT(S): Eight patients with interstitial pregnancy who have undergone laparoscopic cornuotomy. INTERVENTION(S): Laparoscopic cornuotomy was performed using a temporary tourniquet suture and the injection of diluted vasopressin around the cornual mass. The tourniquet suture was removed completely after repairing the cornu. MAIN OUTCOME MEASURE(S): Operating time, hemorrhage, beta-hCG levels. RESULT(S): The estimated blood loss was 50 +/- 22 mL (mean +/- SD), and the operating time was 58 +/- 16 minutes. The serum beta-hCG level returned to within the normal range approximately 4 weeks postoperatively in all patients. There were no major postoperative complications, such as hemorrhage, and no postoperative adjuvant therapy was required. CONCLUSION(S): Laparoscopic cornuotomy is a safe and effective method in interstitial pregnancy, and we believe that it has the advantage of preserving reproductive capacity over cornual resection. PMID- 18394612 TI - Follicular and endocrine response to anastrozole versus clomiphene citrate administered in follicular phase to normoovulatory women: a randomized comparison. AB - OBJECTIVE: To compare the effect on follicular growth and endocrine parameters of follicular phase administration of anastrozole to healthy, normoovulatory women in doses of 1 or 5 mg, respectively, with the conventional dosing regimen for ovulation induction with clomiphene citrate (CC). DESIGN: Randomized, assessor blinded, single-center, three-armed study. SETTING: University-affiliated, tertiary referral center. PATIENT(S): Thirty-two, normoovulatory, normogonadotropic women. INTERVENTION(S): Administration of anastrozole 1 mg (group A1), anastrozole 5 mg (group A5), or CC 50 mg (group CC50) from cycle days 3 to 7. MAIN OUTCOME MEASURE(S): Number of follicles >or=15 mm and >or=10 mm on the day of the endogenous LH surge. RESULT(S): The mean number of follicles >or=15 mm on the day of LH surge was 1.4 +/- 0.5, 1.0 +/- 0.4, and 0.8 +/- 0.4 in groups CC50, A1, and A5, respectively, which was statistically significant. Furthermore, a statistically significant difference between groups was found for the size of the growing follicular cohort (follicles >or=10 mm) on cycle days 8 and 10 and on the day of LH surge. The area under the curve (adjusted for baseline values on cycle day 3 and time frame of assessment) of follicular phase E(2) levels was significantly different among the groups that were compared (223.0 +/- 97.5, 69.2 +/- 40.0, and 83.4 +/- 36.4 for groups CC50, A1, and A5, respectively). CONCLUSION(S): CC 50 mg exerts a stronger stimulatory effect on follicular growth compared with anastrozole in doses of 1 or 5 mg. Anastrozole administration results in lower follicular phase E(2) levels. PMID- 18394614 TI - Tumor necrosis factor alpha and lymphotoxin alpha haplotypes in idiopathic recurrent pregnancy loss. AB - OBJECTIVE: To investigate the contribution of the -238G/A and -308G/A tumor necrosis factor (TNF) alpha, and +252A/G lymphotoxin (LT) alpha gene polymorphisms to idiopathic recurrent miscarriage (RM). DESIGN: A retrospective case-control study. SETTING: Outpatient maternity center. PATIENT(S): Study subjects comprised 372 RM women and 274 age-matched parous control women. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The TNFalpha and LTalpha gene variants and idiopathic RM. RESULT(S): Higher prevalence of TNFalpha -238A and LTalpha +252G alleles and LTalpha +252G/G genotype and lower frequencies of TNFalpha -308G/A were seen in RM cases. Three-loci haplotype analysis (TNFalpha 308GA/TNFalpha -238GA/LTalpha +252AG) demonstrated significant association between TNFalpha-LTalpha gene variants and RM. Both protective [-308A/ 238G/+252A], and susceptible [-308G/-238A/+252G] haplotypes were identified. Mutlivariate regression analysis confirmed the association of -308G/-238A/+252G haplotype with exclusively early RM, after controlling for a number of covariates; no specific TNFalpha and LTalpha genotypes or haplotypes were linked with either late or combined early and late RM. CONCLUSION(S): The TNFalpha 238G/A and LTalpha +252A/G, but not TNFalpha -308G/A, polymorphic variants are associated with exclusively early idiopathic RM. PMID- 18394613 TI - Dioxin may promote inflammation-related development of endometriosis. AB - Laboratory and population-based studies suggest that exposure to environmental toxicants may be one of several triggers for the development of endometriosis. We discuss evidence that modulation of the endometrial endocrine-immune interface could mechanistically link toxicant exposure to the development of this disease. PMID- 18394615 TI - L-carnitine supplementation reduces oocyte cytoskeleton damage and embryo apoptosis induced by incubation in peritoneal fluid from patients with endometriosis. AB - OBJECTIVE: To investigate the protective effect of L-carnitine (LC) against deleterious substances present in the peritoneal fluid (PF) of patients with endometriosis, which may affect the oocyte cytoskeleton and embryogenesis. DESIGN: Experimental study. SETTING: Research embryology laboratory at an academic hospital. PATIENT(S): Frozen metaphase II mouse oocytes and embryos. INTERVENTION(S): One hundred metaphase II mouse oocytes were divided into five groups and incubated: PF from endometriosis patients; PF from endometriosis patients + LC; PF from tubal ligation patients (patient control); LC only; and human tubal fluid (HTF) alone. A total of 180 eight-cell mouse embryos were divided into: endometriosis only; tubal ligation only; endometriosis + LC; LC alone; and HTF alone. MAIN OUTCOME MEASURE(S): Protective effect of LC on oocytes and embryos. RESULT(S): Incubation of the oocytes and the embryos with PF from patients with endometriosis statistically significantly damaged the oocyte microtubules and chromosomes and increased embryo apoptosis compared with controls. Incubation with LC (0.6 mg/mL) statistically significantly improved microtubule and chromosome structure and decreased the level of embryo apoptosis. CONCLUSION(S): We propose the use of LC as a supplement in patients with endometriosis, a novel approach that may help improve in vitro fertilization outcome in these patients. PMID- 18394616 TI - Premature luteinization and in vitro fertilization outcome in gonadotropin/gonadotropin-releasing hormone antagonist cycles in women with polycystic ovary syndrome. AB - OBJECTIVE: To determine the incidence of premature luteinization in patients with polycystic ovary syndrome (PCOS) undergoing controlled ovarian hyperstimulation (COH) with exogenous gonadotropin/GnRH antagonist (GnRH-a); to compare clinical outcomes in patients with and without premature luteinization. DESIGN: Retrospective case series. SETTING: IVF clinic. PATIENT(S): Thirty-five treatment cycles in 30 patients with PCOS. INTERVENTION(S): Controlled ovarian hyperstimulation with gonadotropin/GnRH-a protocol. MAIN OUTCOME MEASURE(S): Premature luteinization defined as a P concentration of >/=1.3 ng/mL on the day of hCG administration; number of oocytes and two pronuclei (2PN) embryos; implantation and clinical pregnancy rates (PR). RESULT(S): The incidence of premature luteinization was 28%. Compared with those without premature luteinization, patients with premature luteinization had a higher number of oocytes retrieved (24.1 +/- 13.3 vs. 12.0 +/- 5.9) and greater number of mature oocytes (19.7 +/- 11.7 vs. 9.5 +/- 4.5), respectively. The number of good quality embryos and embryos transferred was not significantly different between groups. Although implantation rates (56% vs. 40%) and clinical PRs (36% vs. 30%) were higher in patients without premature luteinization, the differences were not statistically significant. CONCLUSION(S): The patients with PCOS with premature luteinization had a higher number of oocytes retrieved and mature oocytes, and similar clinical PRs as patients with PCOS without premature luteinization. PMID- 18394617 TI - Sonohysterography as a first-line evaluation for uterine abnormalities in women with recurrent failed in vitro fertilization-embryo transfer. AB - We evaluated the uterine cavity by sonohysterography (SHG) in patients with recurrent failed IVF-embryo transfer despite transfer of good quality embryos, compared with other diagnostic methods. We found that SHG is highly valuable and should be applied routinely as a first-line, office-based diagnostic tool in such cases. PMID- 18394618 TI - Does vasectomy explain the difference in tubal sterilization rates between black and white women? AB - OBJECTIVE: To examine whether the observed difference in tubal sterilization rates between black and white women is dependent on racial/ethnic differences in vasectomy rates. DESIGN: Secondary analysis of national, cross-sectional survey. SETTING: 2002 National Survey of Family Growth. PATIENT(S): Women 15 to 44 years old with a current partner who were able to provide information about their partner's vasectomy status. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The primary outcome was tubal sterilization. Among women with a current partner who had not undergone vasectomy, a multivariable logistic regression model was used to estimate the effects of race/ethnicity on tubal sterilization after adjusting for potential confounders. RESULT(S): Of the 3,391 women in the sample, 14% of white women had a current partner who had undergone vasectomy compared with 5% of Hispanic women and 4% of black women. Among the 3,064 women whose partners had not undergone vasectomy, black women were more likely to undergo tubal sterilization (odds ratio: 1.6; 95% confidence interval: 1.1, 2.2) on the basis of adjusted multivariable analysis. CONCLUSION(S): After controlling for partner vasectomy status, black women were still more likely to undergo tubal sterilization than white women. PMID- 18394619 TI - Periodontal disease and endometriosis: analysis of the National Health and Nutrition Examination Survey. AB - OBJECTIVE: To investigate whether an association exists between endometriosis and periodontal disease, because endometriosis and periodontal disease are chronic, inflammatory processes more common in patients with systemic autoimmune disorders and because each disease alters immune modulators. DESIGN: Cross-sectional study. SETTING: University health system and statistical center. PATIENT(S): Data for 4136 women, ages 18-50, in the National Health and Nutrition Examination Survey, 1999-2004. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Periodontitis and gingivitis among those patients with and without self-reported endometriosis. RESULT(S): Multinomial logistic regression showed that women with self-reported endometriosis had significantly (57%) higher odds of having both gingivitis and periodontitis relative to not having periodontal disease, compared with women without self-reported endometriosis (adjusted odds ratio, 1.57; 95% confidence interval, 1.06, 2.33), when controlling for other relevant factors. CONCLUSION(S): The results of this study suggest a possible association between endometriosis and periodontal disease. Although it is conceivable that the multifactorial development of endometriosis may be augmented by an immune response to an infectious agent, the potential underlying link between endometriosis and periodontal disease may be a generalized, global immune dysregulation. PMID- 18394620 TI - The use and effectiveness of in vitro fertilization in women with endometriosis: the surgeon's perspective. AB - OBJECTIVE: To assess the use and effectiveness of IVF in a cohort of women undergoing surgery for endometriosis. DESIGN: Cohort study. SETTING: University hospital. PATIENT(S): Four hundred thirty-eight patients who attempted to become pregnant after conservative surgery for endometriosis. INTERVENTION(S): Interview. MAIN OUTCOME MEASURE(S): Pregnancy and IVF use. RESULT(S): One hundred ninety-four women conceived in vivo (44%). One hundred twenty-four women did not undergo IVF despite their infertility status (51% of the group of women who failed to conceive in vivo). One hundred thirty-nine women underwent at least one IVF attempt. The cumulative rate of IVF use at 36 months of infertility was 33%. The live-birth/ongoing pregnancy rate per started cycle and per patient was 10% and 20%, respectively. CONCLUSION(S): In a large tertiary care and referral center, IVF played only a minor role in the treatment of endometriosis-associated infertility. PMID- 18394621 TI - Pregnancy in a hermaphrodite with a male-predominant mosaic karyotype. AB - OBJECTIVE: To report a pregnancy in a hermaphrodite and review of the literature. DESIGN: Case report and literature review. SETTING: Clinical. PATIENT(S): A patient with male-predominant mosaic karyotype 96% 46XY. INTERVENTION(S): Removal of left ovotestis in combination with a supracervical hysterectomy and bilateral salpingo-oophorectomy. MAIN OUTCOME MEASURE(S): Identification of published cases of pregnancy and hermaphroditism. RESULT(S): The current patient had previous pregnancy and a wedge biopsy of her left gonad, which demonstrated an ovotestis and an area suspicious for a gonadoblastoma. After delivery of her second pregnancy, the patient underwent a hysterectomy and bilateral salpingo oophorectomy. The histopathologic evaluation following bilateral oophorectomy demonstrated a residual ovotestis without further evidence of a gonadoblastoma. Review of the literature identified 10 other cases of pregnancy in a hermaphrodite patient. CONCLUSION(S): This is the 11th reported case of fertility in a true hermaphrodite and only the second report of a mosaic true hermaphrodite to demonstrate fertility. This is the only case of a pregnancy involving a male predominant mosaic 96% 46XY and the only case to confirm the genetics of the offspring. PMID- 18394622 TI - Bilateral ovarian pregnancy after intrauterine insemination and controlled ovarian stimulation. AB - OBJECTIVE: To report a case of bilateral ovarian pregnancy in a young patient who had previously undergone intrauterine insemination (IUI) and controlled ovarian stimulation (COS). DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 34-year-old woman who had previously undergone IUI and COS. INTERVENTION(S): Emergency exploratory laparotomy for circulatory collapse. About 350 mL of bloody fluid was collected in the pelvic cavity. The left ovary was about 6 cm in diameter and was completely involved by a darkish hemorrhagic ovarian mass; the right ovary was involved by a 4-cm mass. A left ovariectomy and a partial resection of the right ovary with preservation of two-thirds of ovarian tissue was performed. MAIN OUTCOME MEASURE(S): IUI and COS outcome. RESULT(S): The pathologic diagnosis was considered, and bilateral ovarian pregnancy was confirmed according to the diagnostic criteria described by Spiegelberg. CONCLUSION(S): Diagnosis of primary ovarian pregnancy is very difficult because of its rarity and asymptomatic state before rupture. However, early diagnosis is fundamental to avoid more serious complications and an emergency invasive procedure. PMID- 18394623 TI - Regeneration of spermatogenesis by grafting testicular tissue or injecting testicular cells into the testes of sterile mice: a comparative study. AB - OBJECTIVE: To make a comparison between two different approaches-spermatogonial stem cell transplantation and intratesticular grafting, for preservation and reintroduction of spermatogonial stem cells. DESIGN: Prospective experimental study. SETTING: Academic medical center and teaching hospital. PATIENT(S): N/A. INTERVENTION(S): Intratesticular transplantation, histologic evaluation of testes. MAIN OUTCOME MEASURE(S): Testicular weight, amount of green fluorescence in the testis, and immunostaining for green fluorescent protein. RESULT(S): In a first experiment donor-derived spermatogenesis was found in 65% of the injected testes (41.8 +/- 72.2 mm) compared with 75% of the testes (122.1 +/- 45.6 mm) after tissue grafting. In the second series of experiments complete spermatogenesis was found in 75% of the testes after fresh grafting (93.8 +/- 21.8 mm) compared with 88% after frozen-thawed tissue grafting (84.8 +/- 45.6 mm). CONCLUSION(S): Both approaches show that spermatogonial stem cells can successfully be introduced to the testis resulting in spermatogenesis. Tissue grafting produced a larger mean donor colony length and there was no significant difference between colonization efficiency using either fresh or frozen-thawed grafts. In a future clinical setting, grafting would be a simple and efficient way for reintroducing stem cells to the testis. PMID- 18394624 TI - Comparing the effectiveness of polyethylene covers (Gladwrap) with lanolin (Duratears) eye ointment to prevent corneal abrasions in critically ill patients: a randomized controlled study. AB - BACKGROUND: In unconscious ventilated patients, various eye protective measures have been used to prevent corneal abrasions. Two randomized controlled studies in Australia had compared the effectiveness of polyethylene films and eye instillations to prevent corneal abrasions but results were inconsistent. The local acceptance of polyethylene films as a standard eye protective measure is still limited. OBJECTIVES: Our study aims to compare the effectiveness of polyethylene covers (Gladwrap) with lanolin (Duratears) eye ointment in the prevention of corneal abrasions in critically ill patients. DESIGN: A prospective randomized controlled study was conducted between April 2004 and December 2005. SETTING AND PARTICIPANTS: One hundred and twenty ventilated patients admitted to the intensive care unit (ICU) were randomly assigned to receive either polyethylene covers or lanolin eye ointment to prevent corneal abrasions. METHODS: All participants received a standard eye care regime together with the eye protective interventions. A fluorescein stain test was performed by the eye care team daily and then weekly to detect any corneal abrasions. RESULTS: Four participants were not included in the data analysis as they died soon after commencement of the study. A total of 116 patients were included in the final analysis. Of the seven patients (6.0%) that had a positive fluorescein test, four (6.8%) were in the polyethylene covers group (n=59) and three (5.3%) were in the lanolin eye ointment group (n=57). This was not statistically significant (p=0.519). One patient in the lanolin eye ointment group had an eye infection. Upon follow-up of those patients with positive fluorescein test results, two patients spontaneously converted to stain negative within 24h and two patients died before the ophthalmologist's assessment. The remaining three patients were diagnosed to have epithelial cell loss without corneal abrasions. CONCLUSIONS: With the implementation of a standardized eye care protocol, polyethylene cover is found to be equally effective in preventing corneal abrasions when compared with lanolin eye ointment. The additional benefit of polyethylene cover as a physical barrier to protect patients' eyes needed further evaluation. PMID- 18394625 TI - The mediating effect of job satisfaction between emotional intelligence and organisational commitment of nurses: a questionnaire survey. AB - BACKGROUND: The effect of emotional intelligence and its dimensions on job satisfaction and organisational commitment of nurses has been investigated in this study. OBJECTIVES: This paper examines the relations among emotional intelligence, job satisfaction and organisational commitment of nurses and the mediating effect of job satisfaction between emotional intelligence and organisational commitment. DESIGN: A questionnaire survey was carried out to explore the relations between emotional intelligence, job satisfaction and organisational commitment. SETTING: Teaching hospital in Ankara, Turkey. PARTICIPANTS: Questionnaires were distributed by Nursing Services Administration to 550 nurses working at different departments of the hospital and 267 questionnaires were analyzed. METHODS: A 45-item questionnaire which consists of emotional intelligence, job satisfaction and organisational commitment parts was carried out to investigate the relations among these variables. Some basic socio demographic questions were included. RESULTS: Emotional intelligence was significantly and positively related to job satisfaction (r=0.236,p< or =0.01) and organisational commitment (r=0.229,p< or =0.01). The positive relation between job satisfaction and organisational commitment was also significant (r=0.667,p< or =0.01). Job satisfaction was found to be related with "regulation of emotion (ROE)"(r=0.228,p< or =0.01) and "use of emotion (UOE)"(r=0.155,p< or =0.01) but not with other dimensions of emotional intelligence. "Others's emotional appraisal" did not have any relations with job satisfaction or organisational commitment and "self-emotional appraisal (SEA)" was found to be a suppressor. CONCLUSION: It was found that job satisfaction is a mediator between emotional intelligence and organisational commitment. The other finding of the study was that "SEA" and "UOE" have direct effects on organisational commitment whereas job satisfaction is a mediator between "regulation of emotion" and organisational commitment. PMID- 18394626 TI - Incidence, prevention and treatment of pressure ulcers in intensive care patients: a longitudinal study. AB - BACKGROUND: Pressure ulcers are common in acute and long-term care. However, critically ill patients usually have multiple risk factors for pressure ulcers. OBJECTIVES: The study was conducted to assess pressure ulcer incidence in intensive care patients, the factors related to pressure ulcer incidence and the course of pressure ulcers after the admission to an intensive care unit. DESIGN: A longitudinal design. SETTING: This study was carried out in cardiological and surgical intensive care of a general hospital and in a nephrological intensive care of a university hospital. PARTICIPANTS: All patients admitted to intensive care wards during the period from April until October 2006 were invited to take a part in the study. One hundred and twenty-one patients were involved in the study. The inclusion criteria were adult intensive care patients, males and females, all diagnosis were included. The exclusion criterion was patients whose age less than 18 years. METHOD: Each patient was assessed twice; first, upon admission and second upon discharge or death, or after 2 weeks if the patient was still in intensive care. The assessed data included pressure ulcer preventive measures, risk factors using Braden score, pressure ulcer characteristics and treatment. Additionally, incontinence supplies (urine/bowel) if used and the severity of illness using Acute Physiology and Chronic Health Evaluation (APACHE II score) were assessd. RESULTS: This study revealed a total incidence of 3.3% (4.5% in nephrological patients and 2.9% in surgical patients). Sixteen patients with a total of 21 pressure ulcers were admitted to the intensive care units. During the patients' stay at the intensive care units six pressure ulcers developed newly and five pressure ulcers healed. The mean of the APACHE II score of patients with new pressure ulcers (16.6) were higher than in patients without new pressure ulcers (11.5). CONCLUSION: Pressure ulcer incidence is low in this study compared to other studies. Pressure ulcers can be healed in intensive care patients. Using some preventive measures such as foam and alternating air pressure mattresses may help to decrease pressure ulcer development. Hydrocolloid dressing may help to increase the healing rate of pressure ulcers. PMID- 18394627 TI - Comparison of vessel geometry in bifurcation between normal and diseased segments: intravascular ultrasound analysis. AB - OBJECTIVE: To investigate the geometry in bifurcation using intravascular ultrasound (IVUS) analysis. METHODS: We performed quantitative three-dimensional IVUS analysis of 31 true bifurcation lesions and 30 angiographically normal bifurcation segments including the main and side branches as well as parent vessel. RESULTS: The external elastic membrane (EEM) area changed significantly according to distance from bifurcation in the parent vessel and main branches of the lesion group while there was no significant change in the EEM area in the normal group. Vessel size of bifurcation segments obeyed the principle of minimum work (Murray's law). The cube of the mean EEM diameter of parent vessel nearly equaled the sum of the cubes of the mean EEM diameters of main and side branches (95.5 +/- 38.9 mm(3) versus 90.7 +/- 36.1 mm(3) for the lesion group and 93.4 +/- 49.6 mm(3) versus 85.3 +/- 47.3 mm(3) for the normal group). However, the deviation from the optimal relationship of lumen size was noted more frequently in bifurcation lesion than normal bifurcation segments (71% versus 43%, P=0.03). CONCLUSIONS: The EEM pattern may differ between bifurcation lesions and normal bifurcating segments. Although human coronary artery system obeys the principle of minimum work, the geometry may deviate from the optimal relationship in bifurcation lesions. PMID- 18394628 TI - Invasion of atheromatous plaques into tunica media causes coronary outward remodeling in WHHLMI rabbits. AB - To clarify the mechanism of coronary outward remodeling, we examined atherosclerotic coronary arteries morphologically using WHHLMI rabbits that develop coronary atherosclerosis spontaneously. Perfusion-fixed coronary segments of WHHLMI rabbits were prepared at 500microm intervals. After immunohistochemical staining and histopathological staining, the areas and lengths of the arterial wall and the lesions were measured. Obvious outward remodeling was observed in coronary sections with greater than 40% cross-sectional narrowing. In coronary sections with greater than 40% cross-sectional narrowing, the tunica media was thick at the shoulder of atheromatous plaque and was thin beneath the atheromatous plaques. Macrophages infiltrated those attenuated tunica media expressed matrix metalloproteinases and oxidized LDL was accumulated in those areas. In those areas, collagen fibers and the internal elastic lamina had disappeared partly and apoptotic smooth muscle cells were observed. Proliferation of smooth muscle cells was observed at the attenuated tunica media and adjacent adventitia. The present results suggest that invasion of atheromatous plaques into the tunica media causes coronary outward remodeling. PMID- 18394629 TI - Validation of a non-invasive fluoroscopic imaging technique for the measurement of dynamic knee joint motion. AB - The accurate measurement of the in vivo knee joint kinematics in six degrees-of freedom (6DOF) remains a challenge in biomedical engineering. We have adapted a dual fluoroscopic imaging system (DFIS) to investigate the various in vivo dynamic knee joint motions. This paper presents a thorough validation of the accuracy and repeatability of the DFIS system when used to measure 6DOF dynamic knee kinematics. First, the validation utilized standard geometric spheres made from different materials to demonstrate the capability of the DFIS technique to determine the object positions under changing speeds. The translational pose of the spheres could be recreated to less than 0.15+/-0.09 mm for velocities below 300 mm/s. Next, tantalum beads were inserted into the femur and tibia of two fresh frozen cadaver knees to compare the dynamic kinematics measured by matching knee models to the kinematics from the tantalum bead matching-a technique similar to Roentgen stereophotogrammetric analysis (RSA). Each cadaveric knee was attached to the crosshead of a tensile testing machine and vertically translated at a rate of 16.66 mm/s while images were captured with the DFIS. Subsequently, the tibia was held fixed and the femur manually flexed from full extension to 90 degrees of flexion, as the DFIS acquired images. In vitro translation of the cadaver knee using the tensile testing machine deviated from predicted values by 0.08+/-0.14 mm for the matched knee models. The difference between matching the knee and tantalum bead models during the dynamic flexion-extension motion of the knee was 0.1+/-0.65 degrees /s in flexion speed; 0.24+/-0.16 mm in posterior femoral translation; and 0.16+/-0.61 degrees in internal-external tibial rotation. Finally, we applied the method to investigate the knee kinematics of a living subject during a step ascent and treadmill gait. High repeatability was demonstrated for the in vivo application. Thus, the DFIS provides an easy and powerful tool for accurately determining 6DOF positions of the knee when performing daily functional activities. PMID- 18394631 TI - Predictions of bone remodeling around dental implant systems. AB - This study presents the implementation of a mathematical bone remodeling algorithm to bone adaptation in the premolar area of the mandible around various dental implant systems, and thus sheds a new perspective to the complex interactions in dental implant mechanics. A two-dimensional, plane strain model of the bone was built from a CT-scan. The effect of implant contour on internal bone remodeling was investigated by considering four dental implant systems with contours similar to commercially available ones and another four with cylindrical and conical cross-sections. The remodeling algorithm predicts non-homogeneous density/elastic modulus distribution; and, implant contour has some effect on how this is distributed. Bone density is predicted to increase on the tips of the threads of the implants, but to decrease inside the grooves. Threadless implants favor to develop a softer bone around their periphery, compared to implant systems that have threads. The overall contour (dimensions and the shape) of an implant affect the bone density redistribution, but the differences between different implant systems are relatively small. PMID- 18394630 TI - Impact-induced osteochondral fracture in the tibial plateau. AB - In this study, human tibia plateaus with the meniscus removed were impacted on various regions of the plateau surface via a drop test using a 5mm indenter. Osteochondral blocks containing the failure site were then extracted, chemically fixed, dehydrated, gold-particle coated, and sent for X-ray micro-CT imaging to obtain 3-D image reconstructions of the cartilage and underlying bone. Cartilage failure upon impact appeared to be characteristically brittle in nature. Impacted cartilage from the region not protected by the meniscus showed a relatively large cavernous disruption with microcrack propagation extending radially into the subchondral bone, while impacted cartilage from beneath the meniscus showed less dramatic surface disruption and with no underlying bone failure. PMID- 18394632 TI - Steroid hormone determination in water using an environmentally friendly membrane based extraction technique. AB - In this study, a method was developed for determination of steroid hormones (17beta-estradiol, estrone, 17alpha-ethynylestradiol) in tap and sewage water samples from Sweden. Sample preparation and analysis were performed by a hollow fibre microporous membrane liquid-liquid extraction (HF-MMLLE) set-up combined with gas chromatography-mass spectrometry (GC-MS). In this approach, only the organic liquid in the lumen (10microL) of the hollow-fibre membrane was utilised for depleting extraction. Several parameters were studied, including: type of organic solvent, sample pH, salt and humic acid content. The optimised method allowed the determination of the analyte at the low ngL(-1) level in tap and sewage water. A linear plot gave correlation coefficients better than 0.995 and resulted in a method limit of detection of 1.6, 3 and 10ngL(-1) for 17beta estradiol, estrone, and 17alpha-ethynylestradiol, respectively, in sewage water. Enrichment factors were over 1400 after derivatisation. The repeatabilities at 50 and 600ngL(-1) were better than 10% and 6%, respectively. PMID- 18394633 TI - Development of a method for the quantification of whey allergen traces in mixed fruit juices based on liquid chromatography with mass spectrometric detection. AB - The availability of accurate and sensitive detection methods for food allergens is crucial for the food industry to ensure the correct labelling of their products in order to protect allergic consumers. For this purpose a method using solid-phase extraction and liquid chromatography coupled to mass spectrometry was developed to detect traces of three allergenic cow milk proteins (lactalbumin, lactoglobulins A and B) in mixed-fruit juice samples. Different sample pre treatments were compared and the best recoveries were obtained with a method employing a solid-phase extraction cartridge. Recoveries ranging from 68% to 79% were achieved for 5 and 20microg/ml tested and the limit of detection was set at 1microg/ml. Both full scan and multiple ion monitoring acquisition modes were investigated and compared. The method was utilized to analyse 15 mixed-fruit juices collected from the market and was found to be capable of positively identifying all three milk proteins. The developed method enables the unambiguous determination of allergenic whey proteins in mixed-fruit juices and can assist in the protection of milk allergic individuals. PMID- 18394634 TI - Qualitative evaluation and quantitative determination of 10 major active components in Carthamus tinctorius L. by high-performance liquid chromatography coupled with diode array detector. AB - Flavonoids in the water extract of Carthamus tinctorius L. exhibit potent biological activities such as anti-coagulant, vasodilation, anti-oxidant, neuroprotection and immunosuppressant. A high-performance liquid chromatographic method was established to evaluate the quality of Carthamus tinctorius through a simultaneous quantitation of eight flavonoids, hydroxysafflor yellow A (2), 6 hydroxykaempferol 3,6-di-O-beta-glucoside-7-O-beta-glucuronide (3), 6 hydroxykaempferol 3,6,7-tri-O-beta-glucoside (4), 6-hydroxykaempferol 3-O-beta rutinoside-6-O-beta-glucoside (6), 6-hydroxykaempferol 3,6-di-O-beta-glucoside (7), 6-hydroxyapigenin 6-O-glucoside-7-O-glucuronide (8), anhydrosafflor yellow B (9), and kaempferol 3-O-beta-rutinoside (10), together with two other compounds named guanosine (1) and syringin (5). Among them, compound 8 was identified as a new compound. The compounds were separated on an Alltech Alltima-C(18) column with gradient elution of acetonitrile and 0.01% trifluoroacetic acid. The detection wavelength was 280 nm. All the compounds showed good linearity (r(2) >or= 0.9989). The recoveries, measured at three concentration levels, varied from 94.9% to 105.2%. This method was also validated with respect to precision, repeatability and accuracy, and was successfully applied to quantify the 10 components in 46 batches of C. tinctorius samples from different areas. Significant variations were found in the contents of these compounds in these samples. Compared with the reported analytical methods of C. tinctorius, this simple and reliable method provided a new basis for overall assessment on quality of C. tinctorius and should be considered as a suitable quality control method. PMID- 18394635 TI - Development of a syringe pump assisted dynamic headspace sampling technique for needle trap device. AB - This paper describes a new approach that combines needle trap devices (NTDs) with a dynamic headspace sampling technique (purge and trap) using a bidirectional syringe pump. The needle trap device is a 22-G stainless steel needle 3.5-in. long packed with divinylbenzene sorbent particles. The same sized needle, without packing, was used for purging purposes. We chose an aqueous mixture of benzene, toluene, ethylbenzene, and p-xylene (BTEX) and developed a sequential purge and trap (SPNT) method, in which sampling (trapping) and purging cycles were performed sequentially by the use of syringe pump with different distribution channels. In this technique, a certain volume (1 mL) of headspace was sequentially sampled using the needle trap; afterwards, the same volume of air was purged into the solution at a high flow rate. The proposed technique showed an effective extraction compared to the continuous purge and trap technique, with a minimal dilution effect. Method evaluation was also performed by obtaining the calibration graphs for aqueous BTEX solutions in the concentration range of 1-250 ng/mL. The developed technique was compared to the headspace solid-phase microextraction method for the analysis of aqueous BTEX samples. Detection limits as low as 1 ng/mL were obtained for BTEX by NTD-SPNT. PMID- 18394636 TI - Study of the chemical derivatization of zearalenone and its metabolites for gas chromatography-mass spectrometry analysis of environmental samples. AB - This study compares different silylation procedures of zearalenone and its metabolites: alpha-zearalenol, beta-zearalenol, zearalanone, alpha-zearalanol and beta-zearalanol for gas chromatography-mass spectrometry (GC-MS) analysis. Four silylating agents among the most frequently used to derivatize polar organic compounds were tested: N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), N methyl-N-trimethylsilyltrifluoroacetamide (MSTFA), N,N-diethyltrimethylsilylamine (TMSDEA) and a commercial mixture of N,O-bis(trimethylsilyl)acetamide, trimethylchlorosilane and N-trimethylsilyimidazole. Previous studies showed that the addition of polar and/or basic solvents can significantly improve the yield of a reaction of derivatization. In this work, four solvents were tested: pyridine, dimethylformamide, acetonitrile and acetone. The influence of each solvent was studied as a function of the silylating agent/solvent ratio. The influences of the temperature and of the reaction time on the reaction yields were also evaluated. A GC-MS quantitation method associating methanol chemical ionization and selected ion storage with three ions was developed and successfully tested on a reconstituted sediment spiked in zearalenone and its metabolites. PMID- 18394637 TI - Separation of sanguinarine and chelerythrine in Macleaya cordata (Willd) R. Br. based on methyl acrylate-co-divinylbenzene macroporous adsorbents. AB - Sanguinarine (SAN) and chelerythrine (CHE) are known as major effective components in the quaternary benzo[c]phenanthridine isoquinoline alkaloids (QBA) fraction of Macleaya cordata (Willd) R. Br. but possess different biological activities. In this study, a method for the separation of SAN and CHE based on methyl acrylate-co-divinylbenzene (MA-co-DVB) macroporous adsorbents was established. The relationship between the polarities of the adsorbents and their adsorption-desorption behaviors towards SAN and CHE was investigated. The results showed that, among three selected commercial adsorbents and seven synthesized macroporous polymeric adsorbents with different MA content, the adsorbent No. 5 with 50% MA content provided the best separation power, and the two alkaloids were separated successfully in a gradient eluent process with 60% (v/v) ethanol aqueous and 80% ethanol aqueous contained 8% acetic acid. Dynamic adsorption and desorption tests had been performed in the column packed with the adsorbent No. 5 for optimizing the process parameters. Under the optimized conditions, the ratio of SAN and CHE transformed from 2:1 in the QBA fraction of M. cordata to 1:13 and 25:1 in the products obtained from the two-step gradient elution, and the recoveries of both SAN and CHE were nearly 90%. PMID- 18394638 TI - Use of specific surface areas in inverse gas chromatography studies at zero surface coverage. AB - Inverse gas chromatography (IGC) is frequently used to study adsorption processes at zero surface coverage on microporous activated carbons. This allows to determine the thermodynamic adsorption parameters as equilibrium constants, V(S), standard enthalpies of adsorption, Delta HA degrees, standard free energy of adsorption, Delta GA degrees, and so on. Nevertheless, the surface areas of the adsorbents (microporous carbons in this case) are needed for this purpose. The experimental determination of the surface areas of microporous solids is not univocal and the results depend on the adsorbate employed in the measurements, usually N2 or CO2. This means that the thermodynamic parameters obtained by IGC are subjected to a degree of uncertainty depending on whether N2 or CO2 is used to determine the surface area values. The aim of this paper is to discuss which of the two surface area values is more appropriate to be used in IGC measurements at zero surface coverage. Experimental and theoretical considerations are supplied in a thorough discussion which supports that CO2 surface area value is more appropriate. Thus, it is proposed that this should be used instead of the more generally extended nitrogen specific surface area obtained by the BET equation. PMID- 18394639 TI - The effect of PEO block lengths on the size and stability of complex coacervate core micelles. AB - We report on a series of polyion complexes from mixtures of poly(ethylene oxide) block-poly(N,N-diethylaminoethylmethacrylate) (PEO-PDEAMA) and poly(ethylene oxide)-block-poly(aspartic acid) (PEO-PAsp). As expected, the micelle size, polydispersity and stability are dependant on the relative and absolute lengths of the polyelectrolyte chains. However, we also demonstrate that whilst the length of the charged polyelectrolyte blocks is important, the length of the PEO chains is an equally relevant variable in determining both the size and stability of the final micelles as well as the degree of charge neutralisation at which micellisation occurs. We also show that the kinetics of formation can result in very different stability of the final micelles. PMID- 18394640 TI - Noise levels during aerobics and the potential effects on distortion product otoacoustic emissions. AB - The purpose of this study was to measure noise levels during aerobics classes and to examine how outer hair cell (OHC) function, using distortion product otoacoustic emissions (DPOAEs), may be affected by this exposure. Fifty individuals (48 women and 2 men, ages 19-41 years) participated in 50-min aerobics classes. Noise levels were measured using noise dosimeters placed on the collar near the test ear. The audiometric protocol consisted of a questionnaire, otoscopy, screening tympanometry, and pre- and post-aerobics DPOAEs. The minute by-minute peak noise levels varied between 90.5 and 99.7dBA. The overall mean noise level for the aerobics classes was 87.1dBA (range=83.4-90.7dBA). Mean post aerobic DPOAE levels were lower at most frequencies with a statistically significant 1.4dB decrease at 6000Hz. Results of this project, even with limited statistical significance, should be viewed cautiously. LEARNING OUTCOMES: The reader will be able to (1) identify the approximate dB SPL measured during aerobics classes, (2) describe what effects, if any, these levels had on DPOAEs, and (3) discuss what specific frequency had a significant decrease in DPOAE level after an aerobics class. PMID- 18394641 TI - A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes. AB - Despite their significant role in maintaining the normal physiology, cytokines may cause pathological conditions when they are overproduced. In this way, the increased expression of human interferon alpha (hIFN-alpha) is associated with acute viral infections, inflammatory disorders and several autoimmune illnesses, where the cytokine may be a factor in either initiating or maintaining the disease. Currently, there are several mAbs marketed for a variety of indications and many more in clinical trials, in which IFN-alpha represents a potential target for antibody-based therapy. A panel of 11 murine mAbs was prepared using recombinant hIFN-alpha2b as immunogen, all of which bound to the native form of the cytokine with affinity constants ranging from 1.7x10(7) M(-1) to 1.4x10(10) M(-1). An epitope mapping protocol demonstrated four spatially distinct areas of the protein recognized by the mAbs. Taking into account the characterization of the antibodies and their ability to inhibit the IFN-alpha biological activity, four mAbs were selected to produce scFv fragments. One of these fragments (CA5E6) was able to neutralize a wide spectrum of subtypes of the IFN-alpha family, including the recombinant cytokines hIFN-alpha2a and hIFN-alpha2b and a heterogeneous collection of IFN-alpha produced by activated leukocytes and Namalwa cells. With the aim of improving the affinity of the selected fragment, a standard error-prone PCR method was carried out. By using this strategy, it was possible to generate a new fragment (EP18) with increased affinity and ability to neutralize a broad diversity of IFN-alpha subtypes. Consequently, the scFv EP18 represents a potential therapeutic agent for those immune and inflammatory diseases which are associated with an increased IFN-alpha expression. PMID- 18394642 TI - Infection of Drosophila melanogaster by Tubulinosema kingi: stage-specific susceptibility and within-host proliferation. AB - Despite its importance as a model organism very little is known about the interaction between Drosophila and its microsporidian pathogens. Here we report on the relative susceptibility of Drosophila melanogaster life history stages to infection by Tubulinosema kingi, and on patterns of pathogen proliferation. We find that only larvae can be infected, and that this susceptibility decreases with larval age. Following infection, the pathogen shows little subsequent proliferation in larvae, a limited amount in pupae while it replicates greatly in adults. We present evidence that the host launches a cellular immune response after infection with the pathogen, although its effectiveness remains to be demonstrated. PMID- 18394643 TI - Multifunctional roles of a bacteriophage phi 29 morphogenetic factor in assembly and infection. AB - Low copy number proteins within macromolecular complexes, such as viruses, can be critical to biological function while comprising a minimal mass fraction of the complex. The Bacillus subtilis double-stranded DNA bacteriophage phi 29 gene 13 product (gp13), previously undetected in the virion, was identified and localized to the distal tip of the tail knob. Western blots and immuno-electron microscopy detected a few copies of gp13 in phi 29, DNA-free particles, purified tails, and defective particles produced in suppressor-sensitive (sus) mutant sus13(330) infections. Particles assembled in the absence of intact gp13 (sus13(342) and sus13(330)) had the gross morphology of phi 29 but were not infectious. gp13 has predicted structural homology and sequence similarity to the M23 metalloprotease LytM. Poised at the tip of the phi 29 tail knob, gp13 may serve as a plug to help restrain the highly pressurized packaged genome. Also, in this position, gp13 may be the first virion protein to contact the cell wall in infection, acting as a pilot protein to depolymerize the cell wall. gp13 may facilitate juxtaposition of the tail knob onto the cytoplasmic membrane and the triggering of genome injection. PMID- 18394644 TI - TRPM7 regulates myosin IIA filament stability and protein localization by heavy chain phosphorylation. AB - Deregulation of myosin II-based contractility contributes to the pathogenesis of human diseases, such as cancer, which underscores the necessity for tight spatial and temporal control of myosin II activity. Recently, we demonstrated that activation of the mammalian alpha-kinase TRPM7 inhibits myosin II-based contractility in a Ca(2+)- and kinase-dependent manner. However, the molecular mechanism is poorly defined. Here, we demonstrate that TRPM7 phosphorylates the COOH-termini of both mouse and human myosin IIA heavy chains--the COOH-terminus being a region that is critical for filament stability. Phosphorylated residues were mapped to Thr1800, Ser1803 and Ser1808. Mutation of these residues to alanine and that to aspartic acid lead to an increase and a decrease, respectively, in myosin IIA incorporation into the actomyosin cytoskeleton and accordingly affect subcellular localization. In conclusion, our data demonstrate that TRPM7 regulates myosin IIA filament stability and localization by phosphorylating a short stretch of amino acids within the alpha-helical tail of the myosin IIA heavy chain. PMID- 18394645 TI - Crystal structure of Ca2+ -free S100A2 at 1.6-A resolution. AB - S100A2 is an EF hand-containing Ca(2+)-binding protein of the family of S100 proteins. The protein is localized exclusively in the nucleus and is involved in cell cycle regulation. It attracted most interest by its function as a tumor suppressor via p53 interaction. We determined the crystal structure of homodimeric S100A2 in the Ca(2+)-free state at 1.6-A resolution. The structure revealed structural differences between subunits A and B, especially in the conformation of a loop that connects the N- and C-terminal EF hands and represents a part of the target-binding site in S100 proteins. Analysis of the hydrogen bonding network and molecular dynamics calculations indicate that one of the two observed conformations is more stable. The structure revealed Na(+) bound to each N-terminal EF hand of both subunits coordinated by oxygen atoms of the backbone carbonyl and water molecules. Comparison with the structures of Ca(2+) free S100A3 and S100A6 suggests that Na(+) might occupy the S100-specific EF hand in the Ca(2+)-free state. PMID- 18394646 TI - IL-10 modulates depressive-like behavior. AB - The role of pro-inflammatory cytokines in psychiatric disorders has been the focus of great research attention in recent years. Paradoxically, the same is not true for anti-inflammatory cytokines. In the present study, we assessed the behavioral profile of animals with altered expression of the anti-inflammatory cytokine IL-10. We performed a battery of tests to assess anxiety, depressive like and cognitive behaviors in mice overexpressing IL-10 (PMT10) and IL-10(-/-) animals; in the later mice we also tested the behavioral effect of IL-10 administration. In the forced-swimming test, IL-10(-/-) females displayed increased depressive-like behavior; importantly, this phenotype was reverted by the injection of IL-10. Moreover, mice overexpressing IL-10 presented a decreased depressive-like behavior. Despite the presence of a similar trend, male animals did not reach significant differences in depressive-like behavior. Assessment in the open-field showed that the absence of IL-10 decreased the percentage of time spent in the center of the arena in both male and female mice, while male animals overexpressing IL-10 revealed an opposite behavior. For both sexes, imbalance in IL-10 levels did not affect spatial reference memory. In conclusion, variations in IL-10 expression are associated with an altered depressive-like behavior, but do not influence cognitive performance. Interestingly, IL-10 imbalance produced more profound behavioral changes in females than in male animals. This is in accordance with clinical data demonstrating an increased susceptibility of women to mood disorders, suggesting an interplay between anti-inflammatory cytokines and sexual steroids. PMID- 18394648 TI - Small-for-size syndrome in the rat: does size or technique matter? AB - BACKGROUND: The incidence of the small-for-size syndrome (SFSS) is inversely correlated to the size of the remnant liver or the partial graft. The relevance of factors besides the absolute liver mass is discussed controversially. It is the aim of this study to test the effect of two different mass ligation techniques in comparison with our newly developed parenchyma-preserving vessel oriented liver resection technique on the induction of a SFSS after extended 90% liver resection in the rat. MATERIALS AND METHODS: Ninety percent liver resections were performed using three surgical techniques, two mass ligation techniques, and a vessel oriented technique. Diagnosis of SFSS was based on the combination of biochemical and morphological criteria on the first postoperative day and was related to the outcome on postoperative day 7 and the regenerative capacity of the liver. RESULTS: Only the use of mass ligation techniques was associated with a low 1-wk survival rate (<40%), more pronounced histomorphological signs of liver damage at 24 h postoperatively, and a delayed onset of hepatocyte proliferation. Histological analysis revealed an extended stump necrosis in the paracaval liver and signs of sinusoidal damage in the remaining caudate lobes as morphological correlates of a putative outflow obstruction as the possible underlying reason. The lesions added up to a high small-for-size score in rats operated according to mass ligation techniques. CONCLUSIONS: These results indicate that preservation of functional liver mass and prevention of an outflow obstruction by delicate surgery is essential to prevent a SFSS in a size-reduced liver. PMID- 18394647 TI - Agitated behavior in persons with dementia: the relationship between type of behavior, its frequency, and its disruptiveness. AB - OBJECTIVE: To examine the relationship between type, frequency, and level of disruptiveness of physically aggressive agitated behaviors, physically non aggressive agitated behaviors, verbally aggressive agitated behaviors, and verbally non-aggressive agitated behaviors in persons with dementia. METHODS: The study was conducted in 11 nursing homes in Maryland, one of which housed two participating buildings. Participants were 191 older nursing home residents with dementia. Cohen-Mansfield Agitation Inventory (CMAI) data provided information on frequency and level of disruptiveness of the different types of agitated behavior. RESULTS: The highest overall frequencies were reported for verbal non aggressive behaviors and the highest average disruptiveness was for verbal aggression. Frequency and disruptiveness of behaviors were highly correlated. Disruptiveness was highly correlated across shifts for all syndromes; a difference in magnitude of disruptiveness between shifts was only evident for physically non-aggressive behaviors (such as pacing), which were more disruptive on the day shift than on the evening shift. When controlling for frequency of behavior, physically aggressive behaviors were the most disruptive across both shifts. CONCLUSION: In understanding the impact of agitated behaviors, it is important to take into account both the type of behavior and its frequency. Overall disruptiveness of a type of behavior is different from disruptiveness when the frequency of the behavior is controlled. PMID- 18394649 TI - Estrogen receptor alpha, estrogen receptor beta, and progesterone receptor as possible prognostic factor in radically resected gallbladder carcinoma. AB - BACKGROUND: Gallbladder carcinoma is a relatively rare malignancy with a poor prognosis. We have often encountered patients in whom the course of their disease differed substantially from what would be predicted based on their clinical staging, which highlights the needs to consider additional predictive factors. Gallbladder carcinoma occurs more frequently in women than men, yet expression of the estrogen receptor (ER) and progesterone (PR) have not been studied. We applied an immunohistochemical stain to examine the expression of ER(alpha), ER(beta), and PR in radically resected gallbladder carcinoma. MATERIAL AND METHODS: We immunohistochemically investigated 30 specimens of gallbladder adenocarcinoma tissues using ER(alpha), ER(beta), and PR antibodies. RESULTS: Adenocarcinoma of gallbladder is negative for both ER(alpha) and PR. However, 22 of 30 cases (73.3%) were confirmed positive for ER(beta), which was significantly correlated with tumor differentiation. Five-year survival rates of ER(beta) positive and negative patients were 53.3% and 31.1%, respectively (P = 0.034). In multivariate analysis, only a low proportion score of ER(beta) status was a statistically significant factor (P = 0.033). CONCLUSIONS: Evaluation of ER(beta) expression in gallbladder carcinoma may be an important factor in identifying a poor prognostic group of gallbladder carcinoma. PMID- 18394650 TI - The significance of arginase I administration on the survival of mice bearing NS 1 myeloma cells. AB - BACKGROUND: Arginase I blood levels elevate in cancerous patients and correlate with cancer stages and poor prognosis. Since arginase is capable of enhancing cell growth, it is unclear whether its ominous effect on cancer progression is through the inhibition of immunity or through direct enhancement of cancer cell growth. We tried to clarify this question. METHODS: NS-1 mouse myeloma cells were inoculated intraperitoneally (i.p.) into mice. Purified mouse arginase I was injected daily either intravenously (i.v.) or i.p. for 6 d. A tumor-only control group received i.p. tumor cells without arginase. The survival rates of all mice were recorded. RESULTS: Survival rates were significantly lower in the i.v. group than in the i.p. group (P=0.017) or in the tumor-only control group (P=0.034). As spleen is readily exposed to i.v. arginase, its natural killer cells were studied and were found to have been significantly suppressed by arginase in vitro (P<0.005). CONCLUSION: Our results indicate that the direct inhibition of the immune system by i.v. arginase is more significant in shortening the survival of tumor-bearing mice than localized (i.p.) arginase promotion of tumor cell growth. Thus, an elevation of arginase in a patient's blood is very harmful to the host immune system, e.g. splenic natural killer cells. PMID- 18394651 TI - The neurocognitive enhancement of surgeons: an ethical perspective. AB - Neurocognitive enhancement is a rapidly expanding scientific field. The vast ethical implications of this developing field for surgical practice have yet to be considered within the literature. This article outlines the reasons surgeons may, in the near future, consider using neurocognitive enhancement and addresses the resulting significant ethical implications of this. We do not seek to support or denounce the potential role of neurocognitive enhancement in surgeons, but to stimulate a debate, which, with ever-increasing levels of stimulant use in schools and colleges, and with a pharmaceutical industry driving the creation of new neuroactive products, has now become a necessity. PMID- 18394652 TI - Antiepileptic effects of electroacupuncture vs vagus nerve stimulation on cortical epileptiform activities. AB - Introduced about two decades ago, vagus nerve stimulation (VNS) therapy has been increasingly used for the treatment of refractory epilepsy recently. This study was set out to compare the effects between VNS and electroacupuncture (EA) on pentylenetetrazole (PTZ) induced epileptiform activities in the rat cerebral cortex. Under general anesthesia, the parietal cortex of the rat (n=20) was exposed to record the cortical epileptiform activities. The left vagus nerve was stimulated at 30 Hz, 1 mA or 3 mA for 5 min. For EA, "Dazhui" acupoint (GV14) was stimulated with a pair of acupuncture needles with the same parameters. The results show that both VNS and EA at either 1 mA or 3 mA could inhibit the PTZ induced cortical epileptiform activities, and higher stimulation (3 mA) was not associated with a greater inhibition. In the cases that showed inhibitory responses, there were no statistically significant differences between the two modalities, implying that EA could be comparable to VNS in the treatment of epilepsy. Thus, under current experimental settings, the antiepileptic effect induced by electrical stimulation appeared not vagal specific, and EA could be a good alternative to VNS in the management of epilepsy. PMID- 18394653 TI - Adaptation to the cost of resistance in a haploid clonally reproducing organism: the role of mutation, migration and selection. AB - A model of compensatory evolution with respect to fungicide resistance in a haploid clonally reproducing fungus is developed in which compensatory mutations mitigate fitness costs associated with resistance. The role of mutation, migration and selection in invasion of rare genotypes when the environment changes from unsprayed to sprayed and from sprayed to unsprayed is analysed in detail. In some circumstances (ignoring back mutations) stable internal steady state values for multiple genotypes can be obtained. In these cases a threshold value (f*) for the fraction of the population exposed to the fungicide can be derived for the transition between different steady-state conditions. Conditions are derived for invasion-when-rare of resistant genotypes at boundary equilibria established sometime after the onset of spraying and conversely of sensitive genotypes sometime after the cessation of spraying are derived. In these cases conditions are presented for (a) the invasion of a resistant genotype with a compensatory mutation (resistant-compensated) into a sensitive-uncompensated population that has re-equilibrated following the onset of spraying and (b) the invasion of a susceptible-uncompensated genotype into a resistant-compensated population that has re-equilibrated following the cessation of spraying, provided certain conditions are met. A resistant-compensated genotype may be fixed (or at near-fixation) in the population following a period of spraying, provided the mean intrinsic growth rate of the resistant-compensated genotype in a sprayed environment (over exposed and non-exposed parts of the population) is greater than that of the susceptible-uncompensated genotype. The fraction of the population exposed (the efficiency of spraying) is critical in this respect. However, it is possible for a sensitive-uncompensated genotype to invade provided there is no fitness gain associated with the resistant-compensated genotype, introduction by migration occurs following equilibration of the population to the new environment, and competitive effects are re-imposed when spraying ceases. We further derive a threshold level for the resident resistant-compensated population to reduce to following the cessation of spraying, such that the introduced susceptible-uncompensated genotype will invade. These results will be of use in determining the long-term persistence of resistance in a pathogen population once a fungicide is no longer effective and removed from use. PMID- 18394654 TI - Generation interval contraction and epidemic data analysis. AB - The generation interval is the time between the infection time of an infected person and the infection time of his or her infector. Probability density functions for generation intervals have been an important input for epidemic models and epidemic data analysis. In this paper, we specify a general stochastic SIR epidemic model and prove that the mean generation interval decreases when susceptible persons are at risk of infectious contact from multiple sources. The intuition behind this is that when a susceptible person has multiple potential infectors, there is a "race" to infect him or her in which only the first infectious contact leads to infection. In an epidemic, the mean generation interval contracts as the prevalence of infection increases. We call this global competition among potential infectors. When there is rapid transmission within clusters of contacts, generation interval contraction can be caused by a high local prevalence of infection even when the global prevalence is low. We call this local competition among potential infectors. Using simulations, we illustrate both types of competition. Finally, we show that hazards of infectious contact can be used instead of generation intervals to estimate the time course of the effective reproductive number in an epidemic. This approach leads naturally to partial likelihoods for epidemic data that are very similar to those that arise in survival analysis, opening a promising avenue of methodological research in infectious disease epidemiology. PMID- 18394655 TI - On a temporal model for the Chikungunya disease: modeling, theory and numerics. AB - Reunion Island faced two episodes of Chikungunya, a vector-borne disease, in 2005 and in 2006. The latter was of unprecedented magnitude: one third of the population was infected. Until the severe episode of 2006, our knowledge of Chikungunya was very limited. The principal aim of our study is to propose a model, including human and mosquito compartments, that is associated to the time course of the first epidemic of Chikungunya. By computing the basic reproduction number R(0), we show there exists a disease-free equilibrium that is locally asymptotically stable if the basic reproduction number is less than 1. Moreover, we give a necessary condition for global asymptotic stability of the disease-free equilibrium. Then, we propose a numerical scheme that is qualitatively stable and present several simulations as well as numerical estimates of the basic reproduction number for some cities of Reunion Island. For the episode of 2005, R(0) was less than one, which partly explains why no outbreak appeared. Using recent entomological results, we investigate links between the episode of 2005 and the outbreak of 2006. Finally, our work shows that R(0) varied from place to place on the island, indicating that quick and focused interventions, like the destruction of breeding sites, may be effective for controlling the disease. PMID- 18394656 TI - Chromosomal aberrations in tire plant workers and interaction with polymorphisms of biotransformation and DNA repair genes. AB - We evaluated chromosomal aberrations in lymphocytes of 177 workers exposed to xenobiotics in a tire plant and in 172 controls, in relation to their genetic background. Nine polymorphisms in genes encoding biotransformation enzymes and nine polymorphisms in genes involved in main DNA repair pathways were investigated for possible modulation of chromosomal damage. Chromosomal aberration frequencies were the highest among exposed smokers and the lowest in non-smoking unexposed individuals (2.5+/-1.8% vs. 1.7+/-1.2%, respectively). The differences between groups (ANOVA) were borderline significant (F=2.6, P=0.055). Chromosomal aberrations were higher in subjects with GSTT1-null (2.4+/-1.7%) than in those with GSTT1-plus genotype (1.8+/-1.4%; F=7.2, P=0.008). Considering individual groups, this association was significant in smoking exposed workers (F=4.4, P=0.040). Individuals with low activity EPHX1 genotype exhibited significantly higher chromosomal aberrations (2.3+/-1.6%) in comparison with those bearing medium (1.7+/-1.2%) and high activity genotype (1.5+/-1.2%; F=4.7, P=0.010). Both chromatid- and chromosome-type aberration frequencies were mainly affected by exposure and smoking status. Binary logistic regression analysis revealed that frequencies of chromatid-type aberrations were modulated by NBS1 Glu185Gln (OR 4.26, 95%CI 1.38-13.14, P=0.012), and to a moderate extent, by XPD Lys751Gln (OR 0.16, 95%CI 0.02-1.25, P=0.081) polymorphisms. Chromosome-type aberrations were lowest in individuals bearing the EPHX1 genotype conferring the high activity (OR 0.38, 95%CI 0.15-0.98, P=0.045). Present results show that exposed individuals in the tire production, who smoke, exhibit higher chromosomal aberrations frequencies, and the extent of chromosomal damage may additionally be modified by relevant polymorphisms. PMID- 18394657 TI - Glutamate is critically involved in seizure-induced overexpression of P glycoprotein in the brain. AB - About 30% of patients with epilepsy do not respond adequately to drug therapy, making pharmacoresistance a major problem in the treatment of this common brain disorder. Mechanisms of intractability are not well understood, but may include limitation of antiepileptic drug access to the seizure focus by overexpression of the drug efflux transporter P-glycoprotein (Pgp) at the blood-brain barrier. Increased expression of Pgp has been determined both in epileptogenic brain tissue of patients with intractable epilepsy and in rodent models of temporal lobe epilepsy, including the pilocarpine model. The mechanisms underlying the increase of Pgp after seizures are unclear. We have recently suggested that the excitatory neurotransmitter glutamate, which is excessively released by seizures, is involved in the seizure-induced overexpression of Pgp in the brain. This hypothesis was evaluated in the present study in the pilocarpine model in rats. After 90 min of status epilepticus (SE), diazepam was administered, followed by either vehicle or the glutamate receptor antagonist MK-801 (dizocilpine). Following SE in vehicle treated rats, Pgp expression in brain capillary endothelial cells increased about twofold in the hippocampus, which was completely prevented by MK-801. Furthermore, neurodegeneration developing in the hippocampus and parahippocampal regions was reduced by the glutamate antagonist. In contrast, the Pgp inhibitor tariquidar did not affect the SE-induced overexpression of Pgp or neurodegeneration in most regions examined. The data indicate that seizure-induced glutamate release is involved in the regulation of Pgp expression, which can be blocked by MK-801. The finding that MK-801 counteracts both Pgp overexpression and neuronal damage when administered after SE may offer a clinically useful therapeutic option in patients with refractory SE. PMID- 18394659 TI - The role of the angular gyrus in visual conjunction search investigated using signal detection analysis and transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) delivered over the posterior parietal cortex increases choice reaction times in visual search for a target defined by a conjunction of features. Some recent studies of visual search have taken an approach based on signal detection theory, the findings of which are not addressed by studying the disruptive effects of TMS on reaction time. Here we investigated the role of the posterior parietal cortex in visual search by applying TMS while subjects performed unspeeded feature and conjunction visual search tasks matched for level of difficulty. TMS over the right, but not the left angular gyrus (AG) in the parietal cortex, nor vertex decreased subjects' sensitivity on the conjunction but not the feature search task, as measured by the signal detection measure, d'. Changes in bias, specifically the tendency to make false positive responses, were less clear. We consider the findings in terms of four possible explanation: binding, attentional control, spatial localisation and visuomotor co-ordinate transformations. PMID- 18394658 TI - Blockade of 5-HT 1B receptors facilitates contextual aversive learning in mice by disinhibition of cholinergic and glutamatergic neurotransmission. AB - Serotonergic (5-HT) neurotransmission plays a role in learning and memory processes, but the physiological role of various receptor subtypes is not well characterised. Among these, 5-HT(1B) receptors are located as autoreceptors on 5 HT axons and heteroreceptors on non-serotonergic terminals. This study examined the role of the 5-HT(1B) receptor in one-trial aversive contextual learning using the passive avoidance (PA) task in NMRI mice. Subcutaneous administration of the 5-HT(1B) receptor agonist anpirtoline (0.1-1.0mg/kg) before PA training impaired retention performance 24h later. Combined administration of anpirtoline with the selective 5-HT(1B) receptor antagonist NAS-181 (0.1-1.0mg/kg) fully blocked the impairments. Administration of NAS-181 alone dose-dependently improved PA retention performance. This facilitatory effect was blocked by subthreshold doses of both the muscarinic antagonist scopolamine (0.03 mg/kg) and the NMDA receptor antagonist MK-801 (0.03 mg/kg). NAS-181 also fully blocked the PA impairments induced by an amnesic dose of scopolamine (0.1mg/kg), when administered prior to, but not after, scopolamine. In addition, NAS-181 attenuated PA impairments induced by MK-801 (0.3mg/kg). These findings indicate that 5-HT(1B) receptors are activated at basal levels of 5-HT transmission. The facilitatory effect of NAS 181 involved alleviation of an inhibitory 5-HT tone mediated via 5-HT(1B) receptors on cholinergic and glutamatergic transmission. This disinhibition is expected to occur in neuronal circuits involved in contextual learning including the hippocampus and interconnected cortico-limbic regions. Blockade of brain 5 HT(1B) heteroreceptors may represent a novel therapeutic strategy for restoration of deficient cholinergic and glutamatergic neurotransmission contributing to memory disorders. PMID- 18394660 TI - fMRI evidence for sensorimotor transformations in human cortex during smooth pursuit eye movements. AB - Smooth pursuit eye movements (SP) are driven by moving objects. The pursuit system processes the visual input signals and transforms this information into an oculomotor output signal. Despite the object's movement on the retina and the eyes' movement in the head, we are able to locate the object in space implying coordinate transformations from retinal to head and space coordinates. To test for the visual and oculomotor components of SP and the possible transformation sites, we investigated three experimental conditions: (I) fixation of a stationary target with a second target moving across the retina (visual), (II) pursuit of the moving target with the second target moving in phase (oculomotor), (III) pursuit of the moving target with the second target remaining stationary (visuo-oculomotor). Precise eye movement data were simultaneously measured with the fMRI data. Visual components of activation during SP were located in the motion-sensitive, temporo-parieto-occipital region MT+ and the right posterior parietal cortex (PPC). Motor components comprised more widespread activation in these regions and additional activations in the frontal and supplementary eye fields (FEF, SEF), the cingulate gyrus and precuneus. The combined visuo oculomotor stimulus revealed additional activation in the putamen. Possible transformation sites were found in MT+ and PPC. The MT+ activation evoked by the motion of a single visual dot was very localized, while the activation of the same single dot motion driving the eye was rather extended across MT+. The eye movement information appeared to be dispersed across the visual map of MT+. This could be interpreted as a transfer of the one-dimensional eye movement information into the two-dimensional visual map. Potentially, the dispersed information could be used to remap MT+ to space coordinates rather than retinal coordinates and to provide the basis for a motor output control. A similar interpretation holds for our results in the PPC region. PMID- 18394661 TI - Limited impact of homeostatic plasticity on motor learning in humans. AB - Neuroplasticity is the adaptive modification of network connectivity in response to environmental demands and has been identified as a major physiological correlate of learning. Since unrestricted neuroplastic modifications of network connectivity will result in a de-stabilization of the system, metaplastic modification rules have been proposed for keeping plastic connectivity changes within a useful dynamic range. In this connection, the modification threshold to achieve synaptic strengthening is thought to correlate negatively with the history of activity of the respective neurons, i.e. high previous activity enhances the threshold for synaptic strengthening and vice versa. However, the relevance of metaplasticity for actual learning processes has not been tested so far. We reduced or enhanced motor cortex excitability before performance of the serial reaction time task (SRTT), a sequential motor learning paradigm, and a reaction time task (RTT) by transcranial direct current stimulation (tDCS). If homeostatic rules apply, excitability-diminishing cathodal tDCS should improve subsequent motor learning, especially if combined with the partial NMDA receptor agonist d-cycloserine, which selectively enhances efficacy of active receptors, while excitability-enhancing anodal tDCS should reduce it. Only the results for anodal tDCS, when combined with d-cycloserine, were in accordance with the rules of homeostatic plasticity. We conclude that homeostatic plasticity, as tested here, has a limited influence on implicit sequential motor learning. PMID- 18394662 TI - Endogenous opioids and cannabinoids: system interactions in the regulation of appetite, grooming and scratching. AB - Growing evidence suggests substantial crosstalk between endogenous opioid and cannabinoid systems in the regulation of appetite. Not only is cannabinoid induced hyperphagia abolished by opioid receptor antagonists (and vice versa), but several laboratories have reported supra-additive anorectic responses following co-administration of opioid and CB1 receptor antagonists. In the present study, videoanalysis has been used to characterise the acute effects of sub-anorectic doses of rimonabant (0.25, 0.75 mg/kg) and naloxone (0.1 mg/kg), alone and in combination, on mash intake, ingestive and non-ingestive behaviour, and post-treatment weight gain in male rats. The results confirmed that, when administered alone, none of these treatments significantly altered mash consumption, various measures of feeding behaviour, or weight gain. Although most non-ingestive behaviours were also unaffected, 0.75 mg/kg rimonabant induced compulsive scratching and grooming. However, when naloxone was given in combination with either dose of rimonabant, both food intake and time spent feeding were significantly decreased while the behavioural satiety sequence (BSS) was accelerated. On further analysis, the co-treatment reductions in food intake and feeding behaviour were found to be of an additive rather than supra-additive nature. Intriguingly, the co-administration of naloxone also virtually abolished the compulsive scratching response to the higher dose of rimonabant. Findings are discussed in relation to current views on the molecular bases of opioid cannabinoid system interactions and the unexpected 'dual' advantage (reduction in appetite plus attenuation of side-effect) of low-dose combinations of opioid and cannabinoid CB1 receptor antagonists. PMID- 18394663 TI - Risk factors for active trachoma in children and trichiasis in adults: a household survey in Amhara Regional State, Ethiopia. AB - Identification of risk factors is essential for planning and implementing effective trachoma control programmes. We aimed to investigate risk factors for active trachoma and trichiasis in Amhara Regional State, Ethiopia. A survey was undertaken and eligible participants (children aged 1-9 years and adults aged 15 years and above) examined for trachoma. Risk factors were assessed through interviews and observations. Using ordinal logistic regression, associations between signs of active trachoma in children and potential risk factors were explored. Associations between trichiasis in adults and potential risk factors were investigated using conventional logistic regression. A total of 5427 children from 2845 households and 9098 adults from 4039 households were included in the analysis. Ocular discharge [odds ratio (OR)=5.9; 95% CI 4.8-7.2], nasal discharge (OR=1.6; 95% CI 1.3-1.9), thatch roof in household (OR=1.3; 95% CI 1.0 1.5), no electricity in household (OR=2.4; 95% CI 1.3-4.3) and increasing altitude (Ptrend<0.001) were independently associated with severity of active trachoma. Trichiasis was associated with increasing age (ORper 5 year increase=1.5; 95% CI 1.4-1.7), female gender (OR=4.5; 95% CI 3.5-5.8), increasing prevalence of active trachoma in children (Ptrend=0.003) and increasing altitude (Ptrend=0.015). PMID- 18394664 TI - Mycobacterium tuberculosis resistance in HIV-infected patients from a tertiary care teaching hospital in Porto Alegre, southern Brazil. AB - Drug-resistant Mycobacterium tuberculosis isolates are a major public health concern worldwide. There are few studies assessing tuberculosis (TB) resistance in Brazil. This study assessed the prevalence of resistance to the five first line anti-TB drugs in TB isolates from HIV-infected patients in a tertiary care teaching hospital in southern Brazil. From September 1997 to July 2003, 398 TB complex isolates were included in the study. Resistance to one or more first-line anti-TB drugs was found in 71 (17.8%) of patients and was significantly more frequent among previously treated patients (12 [30.8%] of 39 patients) than new cases (59 [16.4%] of 359) (P=0.05). The highest resistance rates were found to isoniazid (9.9% overall; and 25.6% among previously treated patients). Multidrug resistant TB was found in eight (2.0%) patients, with higher rates among previously treated patients than new cases: two (5.1%) patients vs. six (1.6%), respectively (P=0.18). Multidrug resistance and particularly isoniazid resistance rates among previously treated HIV patients are of great concern. Our findings indicate the need to reappraise regional TB treatment policies and support the recommendation for routine performance of in vitro TB susceptibility tests in all previously treated patients. PMID- 18394665 TI - Homology modelling of human DHCR24 (seladin-1) and analysis of its binding properties through molecular docking and dynamics simulations. AB - Recent biochemical and clinical evidences unveiled that DHCR24 enzyme (3-beta hydoxysterol-Delta(24)-reductase, also named seladin-1), which catalyzes the last step of the cholesterol biosynthesis, is implicated in relevant neuroprotective processes by modulating the level of cholesterol in membrane. The present study was undertaken with a view to model the DHCR24 enzyme and its catalytic site, analyzing the substrate recognition at an atomic level. A homology model of the enzyme was obtained based on plant Cytokinin Dehydrogenase, and its active site was found to bind the desmosterol plus a set of post-squalenic intermediates of the cholesterol biosynthesis in a binding mode conducive to catalysis, even if the docking results suggested that the enzyme has a clear preference for the last intermediates of such biosynthetic pathway. Since DHCR24 possesses a putative transmembrane segment, the enzyme was, then, inserted in a suitable membrane model and the membrane-anchored structure in complex with desmosterol and cholesterol underwent 10ns MD simulations. Such simulations evidenced a clearly different behavior between substrate and product since the product only completely leaves the catalytic cavity whereas desmosterol firmly conserves its pivotal interactions during all simulation time. This is one of the first reports documenting the enzymatic product egress using simple MD simulations in which all atoms are free to move. PMID- 18394666 TI - Simultaneous determination of tetrahydrocortisol, allotetrahydrocortisol and tetrahydrocortisone in human urine by liquid chromatography-electrospray ionization tandem mass spectrometry. AB - Simultaneous quantification method of three major metabolites of cortisone and cortisol, tetrahydrocortisol, allotetrahydrocortisol and tetrahydrocortisone by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI MS/MS) was investigated in a positive mode using a recently developed picolinyl derivatization. Conversion of each steroid into the corresponding picolinyl derivatives (1b, 2b or 3b) was performed by mixed anhydride method using picolinic acids and 2-methyl-6-nitrobenzoic anhydride. Derivatization proceeded smoothly to afford the corresponding 3, 21-dipicolinyl derivatives. Positive ion ESI mass spectra of the picolinyl derivatives were dominated by an appearance of [M+H](+) as base peaks in all cases. The picolinyl derivatives provided 15 to 80 fold higher ESI response in the LC-ESI-MS/MS (selected reaction monitoring: SRM) when compared to those of underivatized molecules in a positive LC-ESI mode. The use of the picolinyl ester, solid-phase extraction, and deuterium labeled internal standards enabled the concentrations of these metabolites in human urine to be determined simultaneously by LC-ESI-MS/MS (SRM) with a small sample volume of less than 1microl urine. PMID- 18394667 TI - Histological evaluation of gonadal differentiation in fathead minnows (Pimephales promelas). AB - The timing of sex determination and the pattern of sex differentiation have not been studied in fathead minnow even though this species of fish are commonly used as a research model for toxicological studies. In this study, the developmental histology of gonadal development was investigated. Fish were cultured in the laboratory conditions and spawning obtained at a photoperiod of 16 h-light and 8 h-dark. Samples were collected from day 7 fish post-spawning (day 7 fps) to day 150 fps and their gonads were processed for histological examination. Developmental histology was assessed by using a light microscopy. The results showed that ovarian differentiation normally occurs at around day 13 fps, while testicular differentiation normally occurs at around day 22 fps. PMID- 18394668 TI - Genome-wide effects of acute progressive feed restriction in liver and white adipose tissue. AB - Acute progressive feed restriction (APFR) represents a specific form of caloric restriction in which feed availability is increasingly curtailed over a period of a few days to a few weeks. It is often used for control animals in toxicological and pharmacological studies on compounds causing body weight loss to equalize weight changes between experimental and control groups and thereby, intuitively, to also set their metabolic states to the same phase. However, scientific justification for this procedure is lacking. In the present study, we analyzed by microarrays the impact on hepatic gene expression in rats of two APFR regimens that caused identical diminution of body weight (19%) but differed slightly in duration (4 vs. 10 days). In addition, white adipose tissue (WAT) was also subjected to the transcriptomic analysis on day-4. The data revealed that the two regimens led to distinct patterns of differentially expressed genes in liver, albeit some major pathways of energy metabolism were similarly affected (particularly fatty acid and amino acid catabolism). The reason for the divergence appeared to be entrainment by the longer APFR protocol of peripheral oscillator genes, which resulted in derailment of circadian rhythms and consequent interaction of altered diurnal fluctuations with metabolic adjustments in gene expression activities. WAT proved to be highly unresponsive to the 4-day APFR as only 17 mRNA levels were influenced by the treatment. This study demonstrates that body weight is a poor proxy of metabolic state and that the customary protocols of feed restriction can lead to rhythm entrainment. PMID- 18394669 TI - An in vitro method to assess toxicity of waterborne metals to fish. AB - The transcription of metal-responsive genes in the rainbow trout (Oncorhynchus mykiss) gill tissue can be used to detect effects of bioreactive metals in natural waters. Here we take advantage of an in vitro gill epithelium, which can be directly exposed to test water samples. The in vitro gill epithelial model mimics the molecular response of in vivo gill epithelial cells to waterborne contaminants. The same culture system can detect trace metals and organic waterborne contaminants. Furthermore, combining this epithelial model with transcriptomic profiling yields an extremely discriminatory biomonitoring tool able to detect and differentiate waterborne metal contaminants. The bioreactive fraction of metal in the water sample is detected using the cells naturally occurring metal sensor, metal-responsive transcription factor 1 (MTF1), which acts upon Metal Response Elements (MRE's) in the enhancer region of metal regulated genes. Induction of the MTF1 responsive genes, metallothionein-A (MTA), metallothionein-B (MTB), and zinc transporter 1 (ZnT-1) in the cell culture was strongly dependent of the concentrations of bioreactive zinc and silver in the test water. Importantly, gene expression in cell culture reflected animal toxicity, measured as inhibition of Ca(2+) and Na(+) influx, in live rainbow trout exposed to the same waters. A cDNA microarray was deployed to determine the differential profiles of transcripts characteristic of exposure to silver, copper or cadmium within this in vitro system. These experiments illustrated the potential power of combining the in vitro gill model epithelium with genetic profiling for accurate characterisation and identification of bioreactive toxicants in waterborne samples. PMID- 18394670 TI - Distinct MAPK signaling pathways, p21 up-regulation and caspase-mediated p21 cleavage establishes the fate of U937 cells exposed to 3-hydrogenkwadaphnin: differentiation versus apoptosis. AB - Despite the depth of knowledge concerning the pathogenesis of acute myeloblastic leukemia (AML), long-term survival remains unresolved. Therefore, new agents that act more selectively and more potently are required. In that line, we have recently characterized a novel diterpene ester, called 3-hydrogenkwadaphnin (3 HK), with capability to induce both differentiation and apoptosis in various leukemia cell lines. These effects of 3-HK were mediated through inhibition of inosine 5'-monophosphate dehydrogenase, a selective up-regulated enzyme in cancerous cells, especially leukemia. However, it remains elusive to understand how cells display different fates in response to 3-HK. Here, we report the distinct molecular signaling pathways involved in forcing of 3-HK-treated U937 cells to undergo differentiation and apoptosis. After 3-HK (15 nM) treatment, a portion of U937 cells adhered to the culture plates and showed macrophage criteria while others remained in suspension and underwent apoptosis. The differentiated cells arrested in G(0)/G(1) phase of cell cycle and showed early activation of ERK1/2 pathway (3 h) along with ERK-dependent p21(Cip/WAF1) (p21) up-regulation and expression of p27(Kip1) and Bcl-2. In contrast, the suspension cells underwent apoptosis through Fas/FasL and mitochondrial pathways. The occurrence of apoptosis in these cells were accompanied with caspase-8-mediated p21 cleavage and delayed activation (24 h) of JNK1/2 and p38 MAPK. Taken together, these results suggest that distinct signaling pathways play a pivotal role in fates of drug-treated leukemia cells, thus this may pave some novel therapeutical utilities. PMID- 18394671 TI - Naturally-occurring estradiol-17beta-fatty acid esters, but not estradiol-17beta, preferentially induce mammary tumorigenesis in female rats: implications for an important role in human breast cancer. AB - Because mammary glands are surrounded by adipose tissues, we hypothesize that the ultra-lipophilic endogenous estrogen-17beta-fatty acid esters may have preferential hormonal and carcinogenic effects in mammary tissues compared to other target organs (such as the uterus and pituitary). This hypothesis is tested in the present study. We found that all 46 rats implanted with an estradiol 17beta pellet developed large pituitary tumors (average weight=251+/-103 mg) and had to be terminated early, but only 48% of them developed mammary tumors. In addition, approximately one-fourth of them developed a huge uterus. In the 26 animals implanted with a mixture containing estradiol-17beta-stearate and estradiol-17beta-palmitate (two representative estradiol-17beta-fatty acid esters) or in the 29 animals implanted with estradiol-17beta-stearate alone (in the same molar dose as estradiol-17beta), 73% and 79%, respectively, of them developed mammary tumors, whereas only 3 or 2 animals, respectively, had to be terminated early due to the presence of a large pituitary tumor. Both tumorous and normal mammary tissues contained much higher levels of estrogen esterase than other tissues, which catalyzes the releases of bioactive estrogens from their fatty acid esters. In conclusion, while estradiol-17beta is much stronger in inducing pituitary tumor (100% incidence) than mammary tumor, estradiol-17beta fatty acid esters have a higher efficacy than estradiol-17beta in inducing mammary tumor and yet it only has little ability to induce uterine out-growth and pituitary tumorigenesis. This study establishes the endogenous estrogen-17beta fatty acid esters as preferential inducers of mammary tumorigenesis. PMID- 18394672 TI - Cytotoxicities of enniatins H, I, and MK1688 from Fusarium oxysporum KFCC 11363P. AB - Enniatins (ENs) H, I, and MK1688 and beauvericin (BEA) were purified from concentrated chloroform extracts of Fusarium oxysporum KFCC 11363P submerged cultures using HPLC, and their in vitro cytotoxicities were evaluated against four human carcinoma cell lines (lung, A549; ovarian, SK-OV-3; skin melanoma, SK MEL-2; and colon, HCT15) using the sulforhodamine B (SRB) method. ENs I and MK1688 inhibited the growth of cancer cell lines most strongly and had similar cytotoxic effects on the tested human cancer cell cultures. The cytotoxicity of ENs I and MK1688 was three- to fourfold higher than that of BEA and EN H. When cultivated in Fusarium-defined medium (FDM), the concentrations of ENs and BEA produced in F. oxysporum KFCC 11363P decreased in the following order: EN MK1688 (0.81 g/L) >EN I (0.55 g/L) >BEA (0.17 g/L) > EN H (0.16 g/L). This study has shown that ENs H, I, and MK1688 exhibit cytotoxicity against certain adenocarcinoma cell lines. The results indicate the need for more investigations into the significance of the biological properties of these new ENs. PMID- 18394673 TI - A two-dimensional CVIB imaging system with a speckle tracking algorithm. AB - Quantitative ultrasound tissue characterization based on integrated backscatter (IB) has shown great potential in detecting myocardial ischemia. The magnitude of the cyclic variation in IB (CVIB) has been considered one promising parameter in assessing regional myocardial contractile performance. This lab has previously developed a novel ultrasonic fusion imaging method based on CVIB. However, the major problem for clinical applications of this technique is that the myocardial tissue could not be tracked effectively without cardiologist's intervention. This paper introduced a speckle tracking method into the CVIB-weighted imaging system, called speckle tracking algorithm with adaptive window size (STAWAWS), to track myocardial tissue particle automatically. This method provides a way to obtain the particle's positions frame by frame in a series of B-mode images. Then using the RF signals according to the particle's positions the IB curve can be calculated to produce CVIB value. The method was applied on the experimental and clinical data cases's analysis. The results of dog's data processing showed that this method could eliminate the misunderstanding of myocardial ischemia especially near the endocardium. The results of clinical data suggested that this method had clinical significance in detecting ischemic myocardium. Though the CVIB-weighted images obtained by the use of this auto-tracking method can improve the accuracy of detecting myocardial ischemia, it is not real-time analysis and the clinical data cases are not sufficient. Further clinical validation is still needed in the future' work. PMID- 18394674 TI - Negative and positive masking responses to light in retinal degenerate slow (rds/rds) mice during aging. AB - Bright light suppresses locomotor activity in mice (negative masking) but dim light augments activity (positive masking). Retinal degeneration slow mice (rds/rds) were tested for responses to light at 3 months, 1 and 2 years old. The suppressive effect of light increased between 1 and 2 years, but the positive response to dim lights was severely reduced at 1 year. No such effects occurred in aging wildtypes. The results indicate that enhancement of negative masking depends on the degree of degeneration of the classical photoreceptors, and that residual function in photoreceptors lacking outer segments is initially sufficient for positive masking. PMID- 18394675 TI - A new nonhydrolyzable reactive cGMP analogue, (Rp)-guanosine-3',5'-cyclic-S-(4 bromo-2,3-dioxobutyl)monophosphorothioate, which targets the cGMP binding site of human platelet PDE3A. AB - The amino acids involved in substrate (cAMP) binding to human platelet cGMP inhibited cAMP phosphodiesterase (PDE3A) are identified. Less is known about the inhibitor (cGMP) binding site. We have now synthesized a nonhydrolyzable reactive cGMP analog, Rp-guanosine-3',5'-cyclic-S-(4-bromo-2, 3 dioxobutyl)monophosphorothioate (Rp-cGMPS-BDB). Rp-cGMPS-BDB irreversibly inactivates PDE3A (K(I)=43.4+/-7.2muM and k(cart)=0.007+/-0.0006 min(-1)). The effectiveness of protectants in decreasing the rate of inactivation by Rp-cGMPS BDB is: Rp-cGMPS (K(d)=72 microM)>Sp-cGMPS (124), Sp-cAMPS (182)>GMP (1517), Rp cAMPS (3762), AMP (4370 microM). NAD(+), neither a substrate nor an inhibitor of PDE3A, does not protect. Nonhydrolyzable cGMP analogs exhibit greater affinity than the cAMP analogs. These results indicate that Rp-cGMPS-BDB targets favorably the cGMP binding site consistent with a docking model of PDE3A-Rp-cGMPS-BDB active site. We conclude that Rp-cGMPS-BDB is an effective active site-directed affinity label for PDE3A with potential for other cGMP-dependent enzymes. PMID- 18394676 TI - Toxicity of the herbicide glyphosate to Chordodes nobilii (Gordiida, Nematomorpha). AB - Nematomorpha (horsehair worms) is a poorly known group of worm-like animals similar to nematodes. Adults are free-living and reproduction takes place in freshwater environments, where preparasitic larvae undergo development. All species have a parasitic juvenil stage and infection may result in the host's death, insects being the most frequent host. Most of the life cycle occurs in freshwater environments, which are often contaminated by different pollutants. Based on the lack of information on the toxicity of herbicides to horsehair worms, the objective of this study is to evaluate the effect of different concentrations of glyphosate (technical grade and formulated product) on Chordodes nobilii (Gordiida, Nematomorpha). Bioassays were performed with embryos and larvae (preparasitic stages), and adults (postparasitic stage). Test organisms were exposed for a short period of time to concentrations ranging between 0.1 and 8 mga.e.l(-1) of glyphosate (technical and formulated). Although embryo development was not inhibited, there was a significant decrease in the infective capacity of larvae derived from eggs that had been exposed to >or= 0.1mg/l. Similar results were obtained for directly exposed larvae. No differences in toxicity were detected between the active ingredient and formulated product. Adult exposed for 96 h to 1.76 mgl(-1) formulated Gly shown a mortality of 50%. Results indicate that C. nobilii is affected at glyphosate concentrations lower than those expected to be found in freshwater environments and those specified in the legislation. PMID- 18394677 TI - Quantitative genetics of modern human cranial variation. PMID- 18394678 TI - Variation in the mandibles from Dmanisi, Georgia. PMID- 18394679 TI - A new archaic Homo sapiens fossil from Lake Eyasi, Tanzania. PMID- 18394680 TI - The role of DNA damage and repair in aging: new approaches to an old problem. AB - DNA damage and mutations have been implicated as key causal events in the biological process of aging. In this context, it has been hypothesized that the complex of genome maintenance systems acts as a longevity assurance system by signaling and repairing damage or removing cells that are beyond repair. In the past, various approaches have been taken to clarify the importance of preserving genome integrity for healthy aging. Here I will briefly review these approaches in the context of the progress made in improving our understanding of the interrelationship between DNA damage, genome maintenance and mutations. PMID- 18394681 TI - Design and evaluation of a filter-based chairside amalgam separation system. AB - This study evaluated the ability of a chairside filtration system to remove particulate-based mercury (Hg) from dental-unit wastewater. Prototypes of the chairside filtration system were designed and fabricated using reusable filter chambers with disposable filter elements. The system was installed in five dental operatories utilizing filter elements with nominal pore sizes of 50 microm, 15 microm, 1 microm, 0.5 microm, or with no system installed (control). Daily chairside wastewater samples were collected on ten consecutive days from each room and brought to the laboratory for processing. After processing the wastewater samples, Hg concentrations were determined with cold vapor atomic absorption spectrometry (USEPA method 7470A). Filter systems were exchanged after ten samples were collected so that all five of the configurations were evaluated in each room (with assignment order balanced by a Latin Square). The numbers of surfaces of amalgam placed and removed per day were tracked in each room. In part two, new filter systems with the 0.5 microm filter elements were installed in the five dental operatories and vacuum levels at the high-velocity evacuation cannula tip were measured with a vacuum gauge. In part three of the study, the chairside filtration system utilizing 0.5 microm and 15 microm filter elements was evaluated utilizing the ISO 11143 testing protocol, a laboratory test of amalgam separator efficiency utilizing amalgam samples of known particle size distribution. Mean Hg per chair per day (no filter installed) was 1087.38 mg (SD = 993.92 mg). Mean Hg per chair per day for the 50 microm, 15 microm, 1 microm, 0.5 microm filter configurations was 79.13 mg (SD = 71.40 mg), 23.55 mg (SD = 23.25 mg), 17.68 mg (SD = 17.35 mg), and 4.25 mg (SD = 6.35 mg), respectively (n = 50 for all groups). Calculated removal efficiencies from the clinical samples were 92.7%, 97.8%, 98.4%, and 99.6%, respectively. ANCOVA on data from the four filter groups, with amalgam-surfaces-removed included as a significant covariate, was statistically significant (P < 0.0001). Tukey post-hoc comparisons (P < or = 0.05) indicated that the 50 microm filter removed less mercury than all other filters and the 0.5 microm removed more mercury than the 50 microm and 15 microm filters. Chairside vacuum measured on chairs with the 0.5 microm filters installed were minimally affected at the time of installation, and then gradually diminished as the filters became loaded with debris. The 0.5 microm configuration passed the ISO 11143 testing protocol at 96.8% efficiency. PMID- 18394682 TI - Environmental and health impact by dairy cattle livestock and manure management in the Czech Republic. AB - In this study we evaluate the potential environmental and health impact of dairy cattle livestock and manure management in the Czech Republic. We present a new approach for national assessments of the environmental impact of an agricultural sector. Emission estimates are combined with a country-specific set of indicators to assess the environmental impact in nine regions with specific environmental characteristics. We estimate the contribution of emissions of ammonia (NH3) and nitrogen oxides (NO) to acidification and terrestrial eutrophication, nitrate (NO3) and phosphate (PO4) to aquatic eutrophication, nitrogen oxides (NO), particulate matter (PM10) and (PM2.5) to human toxicity and methane (CH4) and nitrous oxide (NO) to global warming. We present large regional differences in the environmental and health impact per unit of agricultural production. The regional acidifying, eutrophying and global warming impact of dairy cattle is calculated to be up to three times the national average, depending on the dairy cattle intensity. Aquatic eutrophication is found to be a problem in regions with relatively high eutrophying emissions per hectare of so-called nitrate vulnerable zones. Human toxicity problems caused by dairy cattle livestock and manure management are problematic in regions with a high population density in rural areas. The strength of our approach is the use of country-specific characterisation factors to assess the potential environmental and health impact of agriculture at the sub-national scale. We were able to analyse the potential environmental impact without explicit quantification of specific effects on humans and ecosystems. The results can be used to identify the most polluted areas as well as appropriate targets for emission reduction. PMID- 18394683 TI - Fate and transport of emissions for several trace metals over the United States. AB - A regional model for atmospheric photochemistry and particulate matter is used to predict the fate and transport of five trace metals: lead, manganese, total chromium, nickel, and cadmium over the continental United States during January and July 2001. Predicted concentrations of the metals are compared to observations. Lead predictions have the lowest mean differences with observations and the highest correlation coefficients. They best agree with observations made in January over residential and commercial areas in the eastern United States and worst with observations over remote forests and deserts located in the western United States during July. Manganese predictions show similar abilities to reproduce observations but had larger changes between months. Chromium and nickel predictions show diminishing ability to reproduce observations over both urban and rural areas. Cadmium predictions show the least ability to reproduce observations. Potential causes are examined for the errors in predictions. For errors in lead, manganese and perhaps chromium predictions, aerial suspension and biomass burning are suspected because simulations did not include emissions from these sources. Nickel, cadmium and, to a lower extent, chromium predictions suffer from errors in the emissions that represent current anthropogenic activities. Predicted concentrations of all metals show errors from not including sub-grid processes in meteorological and emission rates. Examples include sea breeze circulation along coastal areas and individual sources in urban areas. These errors reduce the ability to reproduce the time dependence of observations. PMID- 18394684 TI - Factor VII-activating protease in patients with acute deep venous thrombosis. AB - Factor VII-activating protease (FSAP) is involved in haemostasis and inflammation. FSAP cleaves single chain urokinase-type plasminogen activator (scu PA). The 1601GA genotype of the 1601G/A polymorphism in the FSAP gene leads to the expression of a FSAP variant with reduced ability to activate scu-PA, without affecting the ability to activate coagulation Factor VII (FVII). Previous studies have investigated the association of the 1601GA genotype with incidence and progression of carotid stenosis and deep venous thrombosis (DVT). The present study is the first to evaluate the potential association between the FSAP phenotype and DVT. We studied the association between the 1601G/A polymorphism, FSAP activity, FSAP antigen, Factor VIIa (FVIIa), prothrombin fragment 1+2 (F1+2), and C-reactive protein (CRP) in plasmas of 170 patients suspected for DVT. FSAP genotypes were equally distributed in patients with (n=64) and without DVT (n=106), (P=0.94). The 1601GA genotype was associated with significant reduction of FSAP activity (P<0.001) and FSAP antigen levels (P=0.04). Patients with DVT showed significantly higher FSAP activity (P=0.008), FSAP antigen (P=0.003), and F1+2 levels (P<0.001) than patients without DVT. The association between the FSAP measures and DVT disappeared when adjusted for CRP levels. F1+2 correlated positively to FSAP antigen (P=0.01), while FVIIa-levels were comparable in patients with and without DVT. We conclude that even though FSAP measures are significantly increased in patients with acute DVT, alterations in the scu-PA activating properties of FSAP are presumably not markedly involved in the development of acute DVT, and that the association between FSAP and DVT disappears after adjustment for CRP. PMID- 18394685 TI - Impact of left ventricular assist device (LVAD) support on the cardiac reverse remodeling process. AB - With improved technology and expanding indications for use, left ventricular assist devices (LVADs) are assuming a greater role in the care of patients with end-stage heart failure. Following LVAD implantation with the intention of bridge to transplant, it became evident that some patients exhibit substantial recovery of ventricular function. This prompted explantation of some devices in lieu of transplantation, the so-called bridge-to-recovery (BTR) therapy. However, clinical outcomes following these experiences are not always successful. Patients treated in this fashion have often progressed rapidly back to heart failure. Special knowledge has emerged from studies of hearts supported by LVADs that provides insights into the basic mechanisms of ventricular remodeling and possible limits of ventricular recovery. In general, it was these studies that spawned the concept of reverse remodeling now recognized as an important goal of many heart failure treatments. Important examples of myocardial and/or ventricular properties that do not regress towards normal during LVAD support include abnormal extracellular matrix metabolism, increased tissue angiotensin levels, myocardial stiffening and partial recovery of gene expression involved with metabolism. Nevertheless, studies of LVAD-heart interactions have led to the understanding that although we once considered the end-stage failing heart of patients near death to be irreversibly diseased, an unprecedented degree of myocardial recovery is possible, when given sufficient mechanical unloading and restoration of more normal neurohormonal milieu. Evidence supporting and unsupporting the notion of reverse remodeling and clinical implications of this process will be reviewed. PMID- 18394686 TI - Sarcomere mechanics in uniform and non-uniform cardiac muscle: a link between pump function and arrhythmias. AB - Starling's Law and the well-known end-systolic pressure-volume relationship (ESPVR) of the left ventricle reflect the effect of sarcomere length (SL) on stress (sigma) development and shortening by myocytes in the uniform ventricle. We show here that tetanic contractions of rat cardiac trabeculae exhibit a sigma SL relationship at saturating [Ca2+] that depends on sarcomere geometry in a manner similar to skeletal sarcomeres and the existence of opposing forces in cardiac muscle shortened below slack length. The sigma-SL-[Ca2+]free relationships (sigma-SL-CaR) at submaximal [Ca2+] in intact and skinned trabeculae were similar, albeit that the sensitivity for Ca2+ of intact muscle was higher. We analyzed the mechanisms underlying the sigma-SL-CaR using a kinetic model where we assumed that the rates of Ca2+ binding by Troponin-C (Tn C) and/or cross-bridge (XB) cycling are determined by SL, [Ca2+] or stress. We analyzed the correlation between the model results and steady state stress measurements at varied SL and [Ca2+] from skinned rat cardiac trabeculae to test the hypotheses that: (i) the dominant feedback mechanism is SL, stress or [Ca2+] dependent; and (ii) the feedback mechanism regulates: Tn-C-Ca2+ affinity, XB kinetics or, unitary XB-force. The analysis strongly suggests that feedback of the number of strong XBs to cardiac Tn-C-Ca2+ affinity is the dominant mechanism that regulates XB recruitment. Application of this concept in a mathematical model of twitch-stress accurately reproduced the sigma-SL-CaR and the time course of twitch-stress as well as the time course of intracellular [Ca2+]i. Modeling of the response of the cardiac twitch to rapid stress changes using the above feedback model uniquely predicted the occurrence of [Ca2+]i transients as a result of accelerated Ca2+ dissociation from Tn-C. The above concept has important repercussions for the non-uniformly contracting heart in which arrhythmogenic Ca2+ waves arise from weakened areas in cardiac muscle. These Ca2+ waves can reversibly be induced in muscle with non-uniform excitation contraction coupling (ECC) by the cycle of stretch and release in the border zone between the damaged and intact regions. Stimulus trains induced propagating Ca2+ waves and reversibly induced arrhythmias. We hypothesize that rapid force loss by sarcomeres in the border zone during relaxation causes Ca2+ release from Tn-C and initiates Ca2+ waves propagated by the sarcoplasmic reticulum (SR). These observations suggest the unifying hypothesis that force feedback to Ca2+ binding by Tn-C is responsible for Starling's Law and the ESPVR in uniform myocardium and leads in non-uniform myocardium to a surge of Ca2+ released by the myofilaments during relaxation, which initiates arrhythmogenic propagating Ca2+ release by the SR. PMID- 18394687 TI - Clear cell carcinoma of the cervix: a multi-institutional review in the post-DES era. AB - OBJECTIVE: To conduct an outcome analysis of patients with cervical clear cell carcinoma (CCCC) in the post-DES era. METHODS: A retrospective review was conducted at 3 major gynecologic cancer centers of all primary CCCC between 1982 and 2004. RESULTS: CCCC was confirmed in 34 patients. Median age was 53 years. DES exposure was confirmed in 2 (6%) patients. A history of smoking was elicited in 22%, and of abnormal Pap smear 18% patients. Primary surgical resection was performed in all stage I or IIA patients (n=26); they displayed superior 3-year overall survival (OS) compared to advanced stage (n=8) patients (91% vs. 22%, p<0.001). Pelvic lymph node involvement was noted in 25%; all patients with positive para-aortic nodes (20% of patients sampled) had positive pelvic nodes. All node positive patients were treated with adjuvant radiation, but nevertheless displayed reduced progression free (31% vs 92%, p<0.001) and overall survival (80% vs. 100%, p=0.02). Adjuvant radiotherapy did not appear to impact OS in patients with negative lymph nodes. DISCUSSION: This series provides insight into the management of early stage CCCC in the post-DES era. Although these patients may be at slightly higher risk of nodal spread, clear cell histology by itself does not appear to portend a worse prognosis than squamous cell carcinoma of the cervix in the absence of traditional risk factors. Our data suggest that patients with low risk early stage CCCC may be managed with radical surgery alone, without the need for adjuvant chemotherapy or radiation. PMID- 18394689 TI - Fixed-dose rate gemcitabine plus docetaxel as second-line therapy for metastatic uterine leiomyosarcoma: a Gynecologic Oncology Group phase II study. AB - OBJECTIVE: Doxorubicin-based treatment is standard therapy for metastatic uterine leiomyosarcoma. There is no standard second-line therapy. We determined activity of fixed-dose rate gemcitabine plus docetaxel as second-line treatment for metastatic uterine leiomyosarcoma. METHODS: Eligible women with unresectable uterine leiomyosarcoma progressing after prior cytotoxic therapy were treated with gemcitabine 900 mg/m(2) days one and eight over 90 min, plus docetaxel 100 mg/m(2) on day 8 of a 21-day cycle with granulocyte growth factor. Patients with prior pelvic radiation received lower doses. Response Evaluation Criteria in Solid Tumors (RECIST) response was assessed by computed tomography (CT). RESULTS: Forty-eight of 51 women were evaluable for response (one wrong histology, two never treated). Prior therapy was doxorubicin-based in 90%, and ifosfamide-based in 6%. The overall objective response rate is 27%, with complete response in 6.3% (3/48), and partial response in 20.8% (10/48). An additional 50% (24/48) had stable disease (median duration 5.4 months). The median number of cycles per patient was 5.5 (range 1-22); 73% of patients remained progression-free at 12 weeks and 52% at 24 weeks. The predominant toxicity was uncomplicated myelosuppression: thrombocytopenia grade 3 (29%), grade 4 (10.4%); neutropenia grade 3 (12.5%), grade 4 (8.3%) anemia grade 3 (20.8%), grade 4 (4.2%). While pulmonary toxicity was reported, no patient had drug-related pneumonitis/hypoxia type toxicity. Median progression-free survival (PFS) was 5.6+ months (range 0.7 27+ months). The median duration of objective response was 9+ months (range 3.9 24.5+ months). CONCLUSION: Fixed-dose rate gemcitabine plus docetaxel is active second-line therapy for uterine leiomyosarcoma. PMID- 18394692 TI - Randomized trial of a clinic-based, community-supported, lifestyle intervention to improve physical activity and diet: the North Carolina enhanced WISEWOMAN project. AB - OBJECTIVE: To determine if a clinic-based behavioral intervention program for low income mid-life women that emphasizes use of community resources will increase moderate intensity physical activity (PA) and improve dietary intake. METHODS: Randomized trial conducted from May 2003 to December 2004 at one community health center in Wilmington, NC. A total of 236 women, ages 40-64, were randomized to receive an Enhanced Intervention (EI) or Minimal Intervention (MI). The EI consisted of an intensive phase (6 months) including 2 individual counseling sessions, 3 group sessions, and 3 phone calls from a peer counselor followed by a maintenance phase (6 months) including 1 individual counseling session and 7 monthly peer counselor calls. Both phases included efforts to increase participants' use of community resources that promote positive lifestyle change. The MI consisted of a one-time mailing of pamphlets on diet and PA. Outcomes, measured at 6 and 12 months, included the comparison of moderate intensity PA between study groups as assessed by accelerometer (primary outcome) and questionnaire, and dietary intake assessed by questionnaire and serum carotenoids (6 months only). RESULTS: For accelerometer outcomes, follow-up was 75% at 6 months and 73% at 12 months. Though moderate intensity PA increased in the EI and decreased in the MI, the difference between groups was not statistically significant (p=0.45; multivariate model, p=0.08); however, moderate intensity PA assessed by questionnaire (92% follow-up at 6 months and 75% at 12 months) was greater in the EI (p=0.01; multivariate model, p=0.001). For dietary outcomes, follow-up was 90% for questionnaire and 92% for serum carotenoids at 6 months and 74% for questionnaire at 12 months. Dietary intake improved more in the EI compared to the MI (questionnaire at 6 and 12 months, p<0.001; serum carotenoid index, p=0.05; multivariate model, p=0.03). CONCLUSION: The EI did not improve objectively measured PA, but was associated with improved self-reported and objective measures of dietary intake. PMID- 18394691 TI - Curcumin, a constituent of curry, suppresses IgE-mediated allergic response and mast cell activation at the level of Syk. AB - BACKGROUND: Activation of mast cells through the high-affinity receptor for IgE (FcepsilonRI) underlies atopic allergic reactions. Curcumin can block this activation, but the mechanism and the effects of curcumin on IgE-mediated allergic reactions are unknown. OBJECTIVES: We sought to determine the antiallergic activity of curcumin in vivo and its mechanism of action in mast cells. METHODS: The antiallergic activity of curcumin was evaluated in mast cell cultures and the passive cutaneous anaphylaxis model. The effects of curcumin on mast cell signaling events were examined by using immunoblotting, immunoprecipitation, RT-PCR, and other molecular biologic approaches. RESULTS: Curcumin inhibited antigen-mediated activation of mast cells and passive cutaneous anaphylaxis in mice. Suppression of degranulation and secretion of TNF alpha and IL-4 was apparent at concentrations as low as 3 micromol/L curcumin in activated mast cells. Similar concentrations of curcumin suppressed Syk-dependent phosphorylations of the adaptor proteins linker of activated T cells and Grb2 associated binder 2, which are critical for mast cell activation. Although curcumin did not inhibit the phosphorylation of Syk itself, it directly inhibited Syk kinase activity in vitro. Further downstream, activating phosphorylations of Akt and the mitogen-activated protein kinases p38, p44/42 (extracellular signal regulated kinase 1/2), and c-Jun N-terminal kinase, which are critical for the production of inflammatory cytokines, were also inhibited. CONCLUSIONS: Curcumin inhibits Syk kinase-dependent signaling events in mast cells and might thus contribute to its antiallergic activity. Therefore curcumin might be useful for the treatment of mast cell-related immediate and delayed allergic diseases. PMID- 18394693 TI - Valproate affects reproductive endocrine function, testis diameter and some semen variables in non-epileptic adolescent goat bucks. AB - Valproate (VPA) is a major antiepileptic drug with a broad spectrum of antiepileptic activity. There is, however, increasing concern about the possible effects of VPA on reproductive endocrine function. This study investigated the effects of valproate, on the endocrine and reproductive system of adolescent, non epileptic, goat bucks. Nine goat bucks were orally treated with 62.5mg/kg valproate twice daily from 2 to 10 months of age in order to sustain therapeutic plasma concentrations of between 300 and 600 micromol/l. Seven bucks served as controls. Body weights and testicular diameters were recorded. Blood samples were collected for measurement of luteinising hormone (LH), follicle stimulating hormone (FSH) and testosterone three times weekly until sacrifice at approximately 40 weeks of age. Conventional reproductive endpoints were recorded and flow cytometric (FCM) analyses of spermatogenesis, including the sperm chromatin structure were conducted. Valproate-treated bucks had on average a higher body weight, but a lower testis diameter than controls. No significant differences were found for plasma FSH in comparison to controls. Valproate treated bucks differed significantly from the control group by showing lower plasma concentrations of LH and testosterone and a later onset of puberty. A significantly higher proportion of sperm from valproate-treated bucks showed abnormal chromatin, demonstrating a harmful effect on DNA from valproate treatment. These results demonstrate that valproate was able to induce reproductive effects in goat bucks related to the hypothalamic-pituitary-axis, as well as to the testes. PMID- 18394694 TI - Technique sensitivity of water-free one-step adhesives. AB - OBJECTIVES: All-in-one adhesives have been associated with water-related problems, such as phase-separation, polymerization-inhibition and reduced shelf life. In this study we investigated whether these problems could be avoided by a water-free self-etch adhesive that requires a 'wet-bonding' application procedure. Additionally, the technique-sensitivity of such a system was evaluated. METHODS: The micro-tensile bond strength (muTBS) of a water-free and a water-containing experimental one-step adhesive (EXP) bonded to either 'air dried', 'blot-dried' or 'overwet' bur-cut enamel and dentin was determined. Likewise, the muTBS of a commercial water-free adhesive Absolute2 (Denstply Sankin) was determined. The interfacial interaction of the adhesives was evaluated by SEM and TEM. RESULTS: In the blot-dried group, both water-free adhesives obtained the highest bond strength and the bond strength of the water free EXP was comparable to that of the water-containing EXP bonded to an air dried surface. When they were applied to overwet surfaces, the water-free adhesives tended towards a lower bond strength. This was partly attributed to the presence of phase-separation (or 'overwet') droplets in the adhesive layer. Unexpectedly, even on air-dried dentin, the water-free adhesives were able to demineralize dentin, however to a limited extent, hence impairing the bond strength. SIGNIFICANCE: Water-free one-step adhesives are a valuable alternative for conventional water-containing one-step adhesives. However, water-related problems in these adhesives are not ruled out, as they are eventually also mixed with water during the application procedure. As such, they partially fail to meet their objective. In addition, their bonding procedure, which involves 'wet bonding', is more technique-sensitive than dry bonding. PMID- 18394690 TI - Developmental programming and endocrine disruptor effects on reproductive neuroendocrine systems. AB - The ability of a species to reproduce successfully requires the careful orchestration of developmental processes during critical time points, particularly the late embryonic and early postnatal periods. This article begins with a brief presentation of the evidence for how gonadal steroid hormones exert these imprinting effects upon the morphology of sexually differentiated hypothalamic brain regions, the mechanisms underlying these effects, and their implications in adulthood. Then, I review the evidence that aberrant exposure to hormonally-active substances such as exogenous endocrine-disrupting chemicals (EDCs), may result in improper hypothalamic programming, thereby decreasing reproductive success in adulthood. The field of endocrine disruption has shed new light on the discipline of basic reproductive neuroendocrinology through studies on how early life exposures to EDCs may alter gene expression via non-genomic, epigenetic mechanisms, including DNA methylation and histone acetylation. Importantly, these effects may be transmitted to future generations if the germline is affected via transgenerational, epigenetic actions. By understanding the mechanisms by which natural hormones and xenobiotics affect reproductive neuroendocrine systems, we will gain a better understanding of normal developmental processes, as well as develop the potential ability to intervene when development is disrupted. PMID- 18394695 TI - Short-term changes in nematode communities from an abandoned intense sand extraction site on the Kwintebank (Belgian Continental Shelf) two years post cessation. AB - We investigated short-term changes (two years) in nematode communities (density, diversity, biomass and community composition) in an area on the Kwintebank (Belgian Continental Shelf) that was closed for sand extraction activities and compared these patterns to nematode community characteristics from another area on the Kwintebank were sand extraction was still ongoing. Six stations were sampled in 2003 and 2004 and nematode community composition and univariate measures of diversity were compared with values obtained during the extraction period and with a "pre-impact situation" sampled in 1978. Although nematode density, diversity and biomass did not change two years after cessation of the exploitation, nematode community composition did and was more stable than in the extracted site. This is attributed to the absence of continuous disturbances associated with the extraction activities such as the creation and filling up of dredge furrows. As a consequence of the typical life history traits of nematodes, recovery seems to follow different pathways when compared to macrobenthic recovery from the same impact. PMID- 18394696 TI - Single and multi-step carboxymethylation of water yam (Dioscorea alata) starch: synthesis and characterization. AB - Water yam (Dioscorea alata) starch was carboxymethylated by single and multi-step processes for nine times successively. Optimal degree of substitution (DS) of 0.98 and reaction efficiency (RE) of 70.5% were obtained in the single-step carboxymethylation at 40 degrees C, 3 h in isopropanol-water reaction mixture ratio 1:0.16. The ratio of sodium hydroxide and sodium monochloroacetate moles to anhydroglucose unit moles for the optimal DS and RE were 1.62 and 1.39, respectively. The DS increased progressively as the steps of carboxymethylation increased from 2 to 9 and an optimal DS of 2.24 was obtained. Initial increases in carboxymethylation step increased the RE progressively up to 82.1% after the seventh carboxymethylation step but declined with further increases in the carboxymethylation step. The wide angle X-ray diffractometry and the DSC revealed remarkable changes after carboxymethylation because starch crystallinity reduced significantly. Thermogram of native starch showed a characteristic three-step decomposition with 13.16%, 61.54% and 24.79% weight losses progressively while carboxymethyl derivative showed four decomposition stages with 9.86%, 36.57%, 3.04% and 23.07% weight losses progressively. Studies revealed that carboxymethylation improved thermal stability of the native starch. IR spectrometry revealed that carboxymethyl starch showed new bands at nu=1600, 1426 and 1324 cm(-1). The broad band (13)C NMR spectra of the ultrasonically degraded carboxymethyl starch showed a peak at delta=180 ppm which was assigned to carbonyl carbon. PMID- 18394697 TI - Interaction behaviors between chitosan and hemoglobin. AB - The interaction and association between chitosan and hemoglobin (Hb) are studied by UV-vis and fluorescence spectroscopies, viscometry, circular dichroism, dynamic and static light scattering. Chitosan can obviously associate with Hb to form protein-chitosan complexes, which affects microstructure of Hb. The distance between the first association site of chitosan with 214-tryptophan residue in Hb is about 5.473 nm. The intrinsic UV-vis absorption and fluorescence intensities of Hb increase with an increase of chitosan concentration. The alpha-helix in Hb is drawn and changed into beta-sheet. PMID- 18394698 TI - A sandwich structured SiO(2)/cytochrome c/SiO(2) on a boron-doped diamond film electrode as an electrochemical nitrite biosensor. AB - A novel sandwich structured SiO(2) gel/cytochrome c (Cyt c)/SiO(2) gel was designed and constructed on conductive boron-doped diamond (BDD) film substrate. A SiO(2) gel membrane was first in situ deposited on the pretreated positive charged H-terminated BDD electrode with a simple and artful surface vapor sol-gel method. Cyt c was subsequently immobilized onto the SiO(2) membranes by electrostatic attraction, followed by another SiO(2) gel layer in situ depositing on it. The SiO(2) interlayer was conceived to play an important role in the resultant sandwich structured SiO(2)/Cyt c/SiO(2)/BDD electrode as a selective "semi-open" medium, which guaranteed the immobilized Cyt c to maintain high stability and perform good electrochemistry and biocatalysis responses. The bioactivity of Cyt c was well protected and the immobilized biomolecule even didn't denature at extremely high or low pH condition. More attractively, Cyt c in the sandwich structured electrode could be further oxidized into highly reactive Cyt c pi-cation by two-step electrochemical oxidation, which could oxidize NO(2)(-) into NO(3)(-) in the solution. A sensitive determination approach of nitrite was accordingly built up based on this biocatalytic oxidative interaction for the first time and a possible mechanism of the interaction was herein proposed. PMID- 18394699 TI - Influence of cyclic strain and decorin deficiency on 3D cellularized collagen matrices. AB - Cyclic strain evokes the expression of the small leucine-rich proteoglycans decorin and biglycan in 2D cultures and native tissues. However, strain-dependent expression of these proteoglycans has not been demonstrated in engineered tissues. We hypothesized that the absence of decorin may compromise the effect of cyclic strain on the development of engineered tissues. Thus, we investigated the contribution of decorin to tissue organization in cyclically strained collagen gels relative to statically cultured controls. Decorin null (Dcn(-/-)) and wild type murine embryonic fibroblasts were seeded within collagen gels and mechanically conditioned using a Flexcell Tissue Train culture system. After 8 days, the cyclically strained samples demonstrated greater collagen fibril density, proteoglycan content, and material strength for both cell types. On the other hand, increases in cell density, collagen fibril diameter, and biglycan expression were observed only in the cyclically strained gels seeded with Dcn(-/ ) cells. Although cyclic strain caused an elevation in proteoglycan expression regardless of cell type, the type of proteoglycan differed between groups: the Dcn(-/-) cell-seeded gels produced an excess of biglycan not found in the wild type controls. These results suggest that decorin-mediated tissue organization is strongly dependent upon tissue type and mechanical environment. PMID- 18394700 TI - Identification, recombinant production and structural characterization of four silk proteins from the Asiatic honeybee Apis cerana. AB - Unlike silkworm and spider silks assembled from very large and repetitive fibrous proteins, the bee and ant silks were recently demonstrated to consist of four small and non-repetitive coiled-coil proteins. The design principle for this silk family remains largely unknown and so far no structural study is available on them in solution. The present study aimed to identify, express and characterize the Asiatic honeybee silk proteins using DLS, CD and NMR spectroscopy. Consequently, (1) four silk proteins are identified, with approximately 6, 10, 9 and 8% variations, respectively, from their European honeybee homologs. Strikingly, their recombinant forms can be produced in Escherichia coil with yields of 10-60 mg/l. (2) Despite containing approximately 65% coiled-coil sequences, four proteins have very low alpha-helix (9-27%) but unusually high random coil (45-56%) contents. Surprisingly, beta-sheet is also detected in four silk proteins (26-35%), implying the possible presence of beta-sheet in the bee and ant silks. (3) Four proteins lacking of the tight tertiary packing appear capable of interacting with each other weakly but this interaction triggers no significant formation of the tight tertiary packing. The study not only implies the promising potential to produce recombinant honeybee silk proteins for the development of various biomaterials; but also provides the first structural insight into the molecular mechanism underlying the formation of the coiled-coil silks. PMID- 18394701 TI - Pulse treatment with the proteasome inhibitor bortezomib inhibits osteoclast resorptive activity in clinically relevant conditions. AB - Myeloma bone disease is due to bone degradation by osteoclasts, and absence of repair by bone forming osteoblasts. Recent observations suggest that the anti myeloma drug bortezomib, a proteasome inhibitor, stimulates bone formation and may inhibit bone resorption. Here, we tested bortezomib on cultured osteoclasts in conditions mimicking the pulse treatment used in the clinic, thereby avoiding continuous proteasome inhibition and unselective toxicity. A 3 h pulse with 25 nM bortezomib followed by a 3-day culture in its absence markedly inhibited osteoclast activity as evaluated through bone resorption, TRAcP release, and RANKL-induced NF-kappaB translocation into nuclei, an event dependent on proteasomes and critical for osteoclast function. The effect on TRAcP was maximal during the first 24 h post-pulse, and then tended to subside. Importantly, applying this pulse treatment to cultured myeloma cells drastically reduced their survival. We measured next the levels of two bone resorption markers in patients during the 3 days following five and seven therapeutic bortezomib administrations, respectively. These levels decreased significantly already 1-2 days after injection, and then increased, showing temporary inhibition of osteoclast activity and paralleling the in vitro effect on TRAcP. Our study demonstrates a direct inhibition of osteoclasts by bortezomib in conditions relevant to treatment of myeloma. PMID- 18394702 TI - MS-275, a novel histone deacetylase inhibitor with selectivity against HDAC1, induces degradation of FLT3 via inhibition of chaperone function of heat shock protein 90 in AML cells. AB - This study explored the effect of MS-275, a novel histone deacetylase inhibitor (HDACI), against a variety of human leukemia cells with defined genetic alterations. MS-275 profoundly induced growth arrest of acute myelogenous leukemia (AML) MOLM13 and biphenotypic leukemia MV4-11 cells, which possess internal tandem duplication mutation in the fms-like tyrosine kinase 3 (FLT3) gene (FLT3-ITD), with IC50s less than 1 microM, as measured by 3-(4,5 dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay on day two of culture. Exposure of these cells to MS-275 decreased levels of total, as well as, phosphorylated forms of FLT3, resulting in inactivation of its downstream signal pathways, including Akt, ERK, and STAT5. Further studies found that MS-275 induced acetylation of heat shock protein 90 (HSP90) in conjunction with ubiquitination of FLT3, leading to degradation of FLT3 proteins in these cells. This was blunted by treatment with the proteasome inhibitor bortezomib, confirming that FLT was degraded via ubiquitin/proteasome pathway. Moreover, we found that further inhibition of MEK/ERK signaling potentiated the action of MS 275 in leukemia cells. Taken together, MS-275 may be useful for treatment of individuals with leukemia possessing activating mutation of FLT3 gene. PMID- 18394703 TI - Determination of water quality standards for chemical mixtures: extension of a methodology developed for herbicides to a group of insecticides and a group of pharmaceuticals. AB - Monitoring surveys throughout America and Europe have demonstrated the widespread presence of organic synthetic substances such as pesticides and pharmaceuticals in surface water. To avoid deleterious effects on the aquatic system, many countries determine water quality criteria for pesticides. For each substance, the comparison of the concentration measured in water with its criterion gives an indication of the pressure this substance put on the aquatic system. However, in the environment, aquatic organisms are not only exposed to single pesticides but typically to mixtures of these substances. It is therefore of particular importance to take mixtures into account when defining water quality criteria, which is rarely done yet. We recently developed a method to define consistent and comparable water quality criteria for mixtures of herbicides having a similar mode of action. These criteria are calculated based on species sensitivity curves; the method assumes that these curves are parallel for substances having similar mode of action. The aim of this study was to apply our method to six organophosphates (insecticides) and to three beta-blockers (pharmaceuticals), other groups of compounds commonly detected in surface water. We found some evidence that the developed methodology gives consistent results for these groups too. The hypothesis of parallelism was accepted in 2/3 (beta-blockers) and 2/6 of the cases (organophosphates) for both species sensitivity curves based on effect concentrations 50% and on no-observed effect concentrations. The use of water quality criteria for mixture is illustrated by two case studies, which show the importance of taking mixtures into account in water quality legislation. PMID- 18394704 TI - The conceptualization and measurement of quality of life: implications for program planning and evaluation in the field of intellectual disabilities. AB - The concept of quality of life (QOL) is increasingly being used in the field of intellectual disabilities as a conceptual and measurement framework for program planning and evaluation. This article describes the development of a QOL conceptual and measurement framework, and summarizes how this framework is currently being used both nationally and internationally to assess and report personal QOL-related outcomes, to guide quality improvement strategies, and to evaluate the effectiveness of those strategies. Implications of such use are discussed, including those related to understanding mental models, developing internal data systems, supporting organization change, and building on current public policies. The article concludes with reference to the evolving nature of the QOL concept and the impact of this on model development and transdisciplinary research. PMID- 18394705 TI - What does the T-cell receptor recognize when it docks on an MHC-encoded restricting element? AB - The postulate is analyzed that single V-gene segments encode recognition of the allele-specific determinants (a) required for the restrictive response of the alphabeta TCR to peptide. The consequence of this is that the positively selected V-domain, Valpha or Vbeta, engages an allele-specific determinant (a) on one subunit or domain of the MHC-encoded restricting element. The entrained V-domain docks on an invariant determinant (i) on the complementing subunit or domain. Consequently, each functional V-domain expresses an anti-a site and an anti-i site, and all subunits or domains of MHC-encoded restricting elements express an a- and i-determinant. The evidence, both biological and structural, discussed here strongly supports this postulate which has far reaching consequences. PMID- 18394706 TI - Morphologic changes in acute central serous chorioretinopathy evaluated by fourier-domain optical coherence tomography. AB - OBJECTIVE: To investigate morphologic alterations around fluorescein leakage sites using Fourier-domain optical coherence tomography (FD OCT) in acute central serous chorioretinopathy (CSC). DESIGN: Observational case series. PARTICIPANTS: Twenty-one eyes with acute CSC with subjective symptoms for under 3 months. METHODS: Patients underwent measurement of visual acuity, fundus observations, and FD OCT examinations at every visit with the intervals of 2 to 4 weeks until subretinal fluid (SRF) resolved. Fluorescein angiography was performed at baseline to confirm dye leakage sites. Horizontal and vertical OCT scans (B-scans and consecutive raster scans) of the fovea and fluorescein leakage sites were obtained. MAIN OUTCOME MEASURES: Morphologic changes in the retinal pigment epithelium (RPE), detached retina, and subretinal space around the leakage sites were evaluated repeatedly during follow-up. RESULTS: The mean period between baseline and the final examination was 108 days (mean no. of examinations, 3.9). Among 23 leakage sites in 21 eyes, FD OCT showed RPE abnormalities in 22 (96%) sites (14 sites [61%] with a pigment epithelial detachment [PED] and 8 [35%] with a protruding or irregular RPE layer). Fibrinous exudates in the subretinal space and sagging/dipping of the posterior layer of the neurosensory retina above the leakage sites were seen at 12 (52%) and 10 (43%) leakage points, respectively. An RPE defect at the edge of or within the PED was observed in 5 leakage sites (22%); in 2 of these, a defect was detectable after the SRF decreased. The posterior surface of the detached retina was smooth in 17 eyes (81%) and granulated in 4 eyes (19%) (mean duration of subjective symptoms, 10 days and 42 days, respectively). The smooth posterior detached retina became granulated in the presence of residual SRF. A PED remained at the 5 leakage sites in 5 eyes (22%) despite SRF resolution. CONCLUSIONS: Fourier-domain OCT examinations showed detailed morphologic changes in eyes with acute CSC including an RPE defect within the PED at a leakage site through which fluid might pass from the sub-RPE to the subretinal area. Fourier-domain OCT findings may offer new information to facilitate understanding of the mechanisms of acute CSC. PMID- 18394708 TI - The impact of solvent mixtures on neurobehavioral performance: conclusions from epidemiological data. AB - The review of epidemiological studies investigating the neurobehavioral effects of occupational exposure to solvent mixtures sought to contribute to the following issues: (1) Identification of affected cognitive and motor functions. (2) Identification of sensitive neuropsychological tests. (3) Analysis of exposure-effect relationships. The approach was based on the meta-analytical method of effect size estimates. Fifty-three groups from occupational studies were included in the meta-analysis. Forty-eight neuropsychological performance variables could be analyzed as they were included in at least three studies. Seventeen articles provided detailed information on the constituents of mixtures, thereby enabling the computation of an exposure index that allowed the comparison of different mixtures. Significant negative effect sizes were obtained for 12 test variables measuring attention, memory, motor performance and constructional abilities. The greatest proportion of lower performance scores in the exposed groups was shown by different tests of attention: significant effect sizes between d=-0.16 and -0.46 were calculated. Tests of cognitive processing speed, response alternation and inhibition seemed to be sensitive tools for the detection of poorer performance. Exposure-effect relationships were mainly characterized by inconsistent patterns. Crude and inappropriately calculated exposure measures were blamed for this outcome. A healthy worker effect was suggested more consistently: studies examining groups with longer exposure duration obtained smaller effect sizes. Indications of confounding were observed; however, they did not seem sufficient to question consistent effect size patterns. Paying greater attention to the measurement of exposure and including measures of confounding is advisable for future studies and would enhance the explanatory power of cross-sectional studies and meta-analyses. PMID- 18394709 TI - Development of a physiologically based pharmacokinetic and pharmacodynamic model to determine dosimetry and cholinesterase inhibition for a binary mixture of chlorpyrifos and diazinon in the rat. AB - Physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) models have been developed for the organophosphorus (OP) insecticides chlorpyrifos (CPF) and diazinon (DZN). It is anticipated that these OPs could interact at a number of important metabolic steps including: CYP450 mediated activation/detoxification, B esterases [carboxylesterase (CaE), butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE)] or PON-1 (A-esterase) oxon detoxification. We developed a binary PBPK/PD model for CPF, DZN and their metabolites based on previously published models for the individual insecticides. The metabolic interactions (CYP450) between CPF and DZN were evaluated in vitro and suggests that CPF is more substantially metabolized to its oxon metabolite than DZN, which is consistent with observed in vivo potency (CPF>DZN). Each insecticide inhibited the other's in vitro metabolism in a concentration-dependent manner. The PBPK model code used to describe the metabolism of CPF and DZN was modified to reflect the type of CYP450 inhibition kinetics (i.e. competitive vs. non-competitive), while B-esterase metabolism was described as dose-additive, and no PON-1 interactions were assumed between CPF- and DZN-oxon with the enzyme. The binary model was then evaluated against previously published rodent dosimetry and cholinesterase (ChE) inhibition data for the mixture. The PBPK/PD model simulations of the acute oral exposure to single-mixtures (15 mg/kg) vs. binary mixtures (15+15 mg/kg) of CFP and DZN resulted in no differences in the predicted pharmacokinetics of either the parent OPs or their respective metabolites, while cholinesterase inhibition was reasonably described using the dose-additive model. A binary oral dose of CPF+DZN (60+60 mg/kg) did result in observable changes in the DZN pharmacokinetics where C(max) was more reasonably fit by modifying the absorption parameters. It is anticipated that at low environmentally relevant binary doses, most likely to be encountered in occupational or environmental related exposures, that the pharmacokinetics are expected to be linear, and ChE inhibition dose-additive. PMID- 18394707 TI - Immunologic and neurodevelopmental susceptibilities of autism. AB - Symposium 5 focused on research approaches that are aimed at understanding common patterns of immunological and neurological dysfunction contributing to neurodevelopmental disorders such as autism and ADHD. The session focused on genetic, epigenetic, and environmental factors that might act in concert to influence autism risk, severity and co-morbidities, and immunological and neurobiological targets as etiologic contributors. The immune system of children at risk of autism may be therefore especially susceptible to psychological stressors, exposure to chemical triggers, and infectious agents. Identifying early biomarkers of risk provides tangible approaches toward designing studies in animals and humans that yield a better understanding of environmental risk factors, and can help identify rational intervention strategies to mitigate these risks. PMID- 18394711 TI - The H-reflex as a probe: pathways and pitfalls. AB - The Hoffmann (or H) reflex is considered a major probe for non-invasive study of sensorimotor integration and plasticity of the central nervous system in humans. The first section of this paper reviews the neurophysiological properties of the H-reflex, which if ignored create serious pitfalls in human experimental studies. The second section reviews the spinal inhibitory circuits and neuronal pathways that can be indirectly assessed in humans using the H-reflex. The most confounding factor is that reciprocal, presynaptic, and Ib inhibition do not act in isolation during movement. Therefore, characterization of these spinal circuits should be more comprehensive, especially in cases of a neurological injury because neurophysiological findings are critical for the development of successful rehabilitation protocols. To conclude, the H-reflex is a highly sensitive reflex with an amplitude that is the result of complex neural mechanisms that act synchronously. If these limitations are recognized and addressed, the H-reflex constitutes one of the major probes to assess excitability of interneuronal circuits at rest and during movement in humans. PMID- 18394712 TI - Semi-automated quantification of filopodial dynamics. AB - Cellular motility underlies critical physiological processes including embryogenesis, metastasis and wound healing. Nerve cells undergo cellular migration during development and also extend neuronal processes for long distances through a complex microenvironment to appropriately wire the nervous system. The growth cone is a highly dynamic structure that responds to extracellular cues by extending and retracting filopodia and lamellipodia to explore the microenvironment and to dictate the path and speed of process extension. Neuronal responses to a myriad of guidance cues have been studied biochemically, however, these approaches fail to capture critical spatio-temporal elements of growth cone dynamics. Live imaging of growth cones in culture has emerged as a powerful tool to study growth cone responses to guidance cues but the dynamic nature of the growth cone requires careful quantitative analysis. Space time kymographs have been developed as a tool to quantify lamellipodia dynamics in a semi-automated fashion but no such tools exist to analyze filopodial dynamics. In this work we present an algorithm to quantify filopodial dynamics from cultured neurons imaged by time-lapse fluorescence microscopy. The method is based on locating the end tips of filopodia and tracking their locations as if they were free-moving particles. The algorithm is a useful tool and should be broadly applicable to filopodial tracking from multiple cell types. PMID- 18394713 TI - Impact of maternal psychological distress on fetal weight, prematurity and intrauterine growth retardation. AB - BACKGROUND: There are conflicting results regarding the association of maternal antenatal distress with preterm birth and low birth weight. This study investigated the association between maternal distress and intrauterine growth abnormality, low birth weight and preterm birth. METHODS: Three mutually exclusive and homogeneous groups of pregnant women (with actual psychiatric disorder, with maternal psychological distress, and healthy comparisons) underwent fetal ultrasound examinations, uterine and umbilical artery Doppler velocimetry. Infant weight was measured and information collected on obstetrical features and sociodemographic factors. RESULTS: No differences emerged among the three groups of pregnant women in any ultrasound variables. Antenatal maternal psychiatric disorders and antenatal distress were not associated with an increased risk of preterm birth. Infants of women with psychiatric disorders had lower birth weight and higher percentage of birth weight below the 10th centile for gestational age (30%) than infants of healthy mothers (5%). LIMITATIONS: These findings are preliminary and warrant further investigation in larger-scale study; they are limited by the heterogeneity of psychiatric diagnoses. CONCLUSIONS: Maternal psychiatric disorders are associated with a lower birth weight, but the effect is unlikely to be due to abnormal utero-placental or feto placental vascularisation. Further studies should investigate other possible causes of lower birth weight associated with maternal psychiatric disorders. PMID- 18394714 TI - Social class, family history and type of schizophrenia. AB - To date, there are numerous studies supporting a genetic model of schizophrenia. There is a paucity of studies, however, screening for a connection between family history of serious mental illness and deficit vs. nondeficit schizophrenia. The aim of the present study was to explore the association between family history, deficit vs. nondeficit schizophrenia and socioeconomic status (SES) of family of origin. Patients (N=437) from a United States psychiatric hospital were separated into deficit vs. nondeficit presentation and bifurcated into poor vs. nonpoor SES. Family history data were utilized to classify patients into subgroups characterized by serious mental illness within immediate family, within extended family, or no evidence of mental illness. Statistical testing was conducted using logistic regression analysis. SES of family of origin was significantly associated with schizophrenic subtype independently of family history, sex and race; specifically, poverty raised the risk of deficit schizophrenia. Family history of mental illness showed no net association, and no statistical interaction with poverty, in predicting risk of deficit schizophrenia. PMID- 18394710 TI - Ethanol-BDNF interactions: still more questions than answers. AB - Brain-derived neurotrophic factor (BDNF) has emerged as a regulator of development, plasticity and, recently, addiction. Decreased neurotrophic activity may be involved in ethanol-induced neurodegeneration in the adult brain and in the etiology of alcohol-related neurodevelopmental disorders. This can occur through decreased expression of BDNF or through inability of the receptor to transduce signals in the presence of ethanol. In contrast, recent studies implicate region-specific up-regulation of BDNF and associated signaling pathways in anxiety, addiction and homeostasis after ethanol exposure. Anxiety and depression are precipitating factors for substance abuse and these disorders also involve region-specific changes in BDNF in both pathogenesis and response to pharmacotherapy. Polymorphisms in the genes coding for BDNF and its receptor TrkB are linked to affective, substance abuse and appetitive disorders and therefore may play a role in the development of alcoholism. This review summarizes historical and pre-clinical data on BDNF and TrkB as it relates to ethanol toxicity and addiction. Many unresolved questions about region-specific changes in BDNF expression and the precise role of BDNF in neuropsychiatric disorders and addiction remain to be elucidated. Resolution of these questions will require significant integration of the literature on addiction and comorbid psychiatric disorders that contribute to the development of alcoholism. PMID- 18394716 TI - Toll-like receptor gene expression in cecum and spleen of advanced intercross line chicks infected with Salmonella enterica serovar Enteritidis. AB - Understanding the changes in host gene expression that occur with bacterial infection will help to elucidate the basis of molecular genetic control of disease resistance. The effect of infecting chicks with Salmonella enterica serovar Enteritidis on the RNA expression level of Toll-like receptor (TLR) genes, and the correlation between TLR RNA expression level and bacterial burden in the cecum and spleen of young birds was studied. Chicks from two advanced intercross lines were either infected or mock infected with S. enteritidis at 1 day of age. The RNA expression levels of TLR2, TLR4 and TLR5 genes were assessed by quantitative reverse transcriptase-PCR (qRT-PCR) in cecum and spleen tissues harvested at one week post-infection. Infected chicks had significant upregulation of TLR2 RNA expression in spleen, TLR4 RNA expression in both cecum and spleen, and downregulation of TLR5 RNA expression in cecum. Bacterial burden of S. enteritidis in infected birds was not correlated with TLR RNA expression level. Infecting chicks with S. enteritidis caused an increase in TLR2, TLR4 and TLR5 RNA expression level in spleen in males but not in females. The effect of sex on response to S. enteritidis infection suggests a role for TLR signaling pathways in sex-based modulation of immune response to pathogens. High correlation between TLR2 and TLR4 mRNA expression level in cecum of S. enteritidis infected birds suggests coordinated regulation or simultaneous stimulation of these genes by S. enteritidis. In conclusion, this study clearly showed that young chicks respond to S. enteritidis infection by upregulating TLR2, TLR4 RNA expression. The downregulation of TLR5 RNA expression was observed in cecum by S. enteritidis infection, which might be beneficial to protect host cells from overstimulation by bacterial flagellin. PMID- 18394715 TI - Transfer of maternal colostral leukocytes promotes development of the neonatal immune system Part II. Effects on neonatal lymphocytes. AB - It has been established that maternal leukocytes, conditioned by the mammary environment, cross the neonatal gut and circulate in the newborn calf. However, the impact of these cells on the development of neonatal immunity remains to be determined. This study examined the effects of maternal colostral leukocytes on development and maturation of neonatal adaptive immunity by examining the expression of surface markers on neonatal lymphocytes. At birth, neonatal calves were fed whole colostrum, or colostrum that had the maternal cells removed (cell free colostrum), from their respective dams. Peripheral blood samples were collected at regular intervals over the first 4 weeks of life and lymphocytes were evaluated for surface expression of cellular markers. The results of these studies demonstrated that calves receiving whole colostrum had fewer CD11a positive lymphocytes in circulation during the first 2 weeks of life and this marker was expressed at a lower density than calves receiving cell-free colostrum. In addition, calves receiving whole colostrum also had a higher percentage of lymphocytes expressing the activation markers CD25 and CD26 by 7 days after birth. During the first week of life, lymphocytes from calves receiving whole colostrum had a higher density of MHC class I expression on their surfaces than cells from calves receiving cell-free colostrum. In general, these results indicate that transfer of maternal cells with colostrum allows for more rapid development of lymphocytes and maternal cells appeared to enhance their activation. PMID- 18394717 TI - Increased expression of the peroxisome proliferator-activated receptor gamma in the immune system of weaned pigs after Escherichia coli lipopolysaccharide injection. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma), a member of the nuclear hormone receptor superfamily, has been implicated in regulation of immunity and inflammation in rodents and humans. The objective of the current study was to investigate whether the expression of PPARgamma was altered in the immune system of weaned pigs after Escherichia coli lipopolysaccharide (LPS) injection. PPARgamma expression was investigated in the thymus, spleen, mesenteric lymph node and peripheral white blood cells of weaned pigs (8.54+/ 0.24 kg BW) after LPS injection (100 microg/kg BW, n=6) and controls (sterile saline, n=6), by using real-time polymerase chain reaction, Western blot analysis, and immunohistochemistry. Plasma pro-inflammatory cytokines and hormones were also assessed. LPS triggered PPARgamma mRNA and protein expression in the thymus (P<0.05, 4.24-fold; P<0.10, 1.46-fold), spleen (P<0.10, 2.75-fold; P<0.05, 1.84-fold), mesenteric lymph node (P<0.05, 4.32-fold; P<0.05, 1.96-fold) and peripheral white blood cells (P<0.001, 24.44-fold; P<0.001, 1.58-fold). The LPS-injected pigs showed an increase in PPARgamma staining in splenic corpuscle and periarterial lymphatic sheath of white pulp (P<0.05) and red pulp (P<0.001) of spleen, and in medullas of thymus lobule of thymus (P<0.05), and in thymus dependent area of mesenteric lymph node (P<0.05) compared to the control pigs. Concurrent with up-regulation of PPARgamma expression, LPS induced increases in plasma interleukin-6 (P<0.001), tumor necrosis factor-alpha (P<0.001), cortisol (P<0.001), prostaglandin E(2) (P<0.01) and 15-deoxy-Delta(12,14)-prostaglandin J(2) (15 d-PGJ(2)) (P<0.05), and decreases in plasma insulin (P<0.10) and insulin like growth factor-1 (P<0.001). These results suggest that induction of PPARgamma expression in immune system may be associated with the release of the natural PPARgamma activating ligand 15 d-PGJ(2), and play an important role in host response to immunological stress. Additionally, it is possible that PPARgamma would be a new therapeutic target in treatment of immunological stress of livestock. PMID- 18394718 TI - The effect of stapes mobility on hearing outcome and which procedure to choose in fixed stapes in children tympanosclerosis. AB - OBJECTIVES: Our objective was to evaluate the features of tympanosclerosis in children and to determine the effect of stapes mobility and the type of one-stage operation on hearing outcomes. MATERIALS AND METHODS: Fifty-one children who were performed different types of single-stage otologic surgery for tympanosclerosis between January 1997 and December 2006 were retrospectively chart reviewed. The children were divided into two groups according to the mobility of ossicular chain, especially the stapes. Stapes fixed group was also evaluated in detail according to the type of surgery that was performed. Patients who had previous ventilation tube insertion, tympanic membrane parasynthesis or any other otologic surgery were excluded from the study. Improvement of the hearing by at least 10 dB and air-bone gap less than 20 dB were accepted as success criteria after 24 months of follow-up period. RESULTS: The air conduction levels, and the air-bone gap values of both groups were improved significantly after the single-stage operations. Pure tone averages pre- and postoperatively for stapes mobile group were 45.55+/-15.96 and 34.50+/-16.64 dB (p=0.002); and in stapes fixed group these were respectively 43.97+/-13.45 and 33.16+/-12.14 dB (p<0.001). When pre- and postoperative air-bone gap levels were evaluated it was seen that in both groups they were improved more than 10 dB, from 34.10+/-11.37 to 23.05+/-12.32 dB (p=0.002) in stapes mobile group and from 35.29+/-11.65 to 24.48+/-12.50 dB (p<0.001) in stapes fixed group. In stapes fixed group air-bone gap was less than 20 dB in 11 of 23 (47.8%) patients who had mobilization and 3 of 8 (37.5%) patients who had small fenestra stapedotomy operations. Although it was not statistically significant, gain was more than 10 dB only in 2 of 8 (25.0%) patients in the stapedotomy group but 14 of 23 (60.9%) patients in mobilization group (p=0.698 for ABG and p=0.220 for gain). The change in the bone conduction levels were improved 0.75 dB in group 1 and got worse 0.52 dB in group 2 and this was not statistically significant (p=0.239). CONCLUSIONS: In this study about children, the status of stapes and the place of tympanosclerotic mass had no significant negative effect on hearing improvement. You can perform mobilization in one-stage if you are experienced and have to prefer second-stage surgery if stapes is fixed and stapedectomy is needed. PMID- 18394719 TI - The relationship between anatomical variations of the sino-nasal region and chronic sinusitis extension in children. AB - OBJECTIVE: To report the prevalence of paranasal sinus anatomic variations and their relationship with the extent of chronic sinusitis in pediatric population. METHOD: This is a prospective study of all the cases of persistent pediatric chronic sinusitis despite medical therapy that presented to the otolaryngology clinics of King Abdullah University Hospital in Irbed, Jordan, and had coronal computed tomography examination of the nose and paranasal sinuses as part of their diagnostic or preoperative work up, between the periods of April 2006 and August 2007. Sixty-five cases met inclusion criteria and were analyzed. Cases were further subdivided into three groups according to age. Group 1: with ages from 5 to 7 years; group 2: ages 8-12; and group 3: above 12 years. The scans were analyzed for the presence of anatomical variations, and disease extension. Correlation between anatomical variations and disease extension was the primary outcome measure. RESULTS: There were 30 boys and 35 girls with an average age of 11.4 (age ranges from 5 to 16). The maxillary sinus was the most commonly involved sinus, followed by the ethmoid, sphenoid, and frontal sinuses. Agger nasi cell was the most common anatomical variation, followed by concha bullosa of middle turbinate. The frequency of most of these variants differed widely between groups 1 and 2, groups 1 and 3, but not between groups 2 and 3. No significant correlation was found between these anatomical variants and the extent of chronic sinusitis. CONCLUSION: Anatomical variations in the nose and paranasal sinuses are common in children with chronic sinusitis. Although the prevalence of these variations changes during childhood development they are unlikely to predict the extent and severity of chronic rhinosinusitis in pediatric age group. PMID- 18394720 TI - FAAH and anandamide: is 2-AG really the odd one out? AB - Fatty acid amide hydrolase (FAAH) is a hydrolytic enzyme that recognizes as substrates and inactivates the two most studied endocannabinoids, anandamide and 2-arachidonoylglycerol (2-AG). Following the observation that endocannabinoids produced by tissues during pathological conditions often have protective roles, FAAH inhibitors have been proposed as therapeutic drugs. Yet it has been suggested that FAAH functions in vivo only as an anandamide-degrading enzyme because its pharmacological and genetic inactivation is usually accompanied by elevation of anandamide, but not 2-AG, levels. We believe, however, that this concept needs to be revisited in light of reports that, under certain experimental conditions, FAAH inhibitors also elevate 2-AG tissue levels in vivo and, more recently, that FAAH inactivation in the striatum instead reduces 2-AG concentrations through upregulation of anandamide levels, activation of transient receptor potential vanilloid 1 receptors and inhibition of 2-AG biosynthesis. PMID- 18394721 TI - Drugging the Plasmodium kinome: the benefits of academia-industry synergy. AB - Malaria remains a major killer in many parts of the world. Recently, the development of nonprofit organisations aimed at fighting this deadly scourge incited academic and industrial scientists to merge their expertise in drug target validation and lead discovery. Expectations are clear: identification and characterisation of new molecules showing high efficacy, low toxicity and little propensity to induce resistance in the parasite. In this context, protein kinase inhibitors represent an attractive possibility. Here, we compare traditional target-based drug-discovery approaches with innovative exploratory paths (parallel screening, cell-based assays, integrated systems biology and allosteric inhibition) and discuss the benefits of acadaemia-industry cooperation. Early characterisation of distribution, metabolism, pharmacokinetic (DMPK) and toxicology parameters are considered as well. PMID- 18394722 TI - Construction and immunogenicity of recombinant pseudotype baculovirus expressing the capsid protein of porcine circovirus type 2 in mice. AB - Baculovirus has emerged recently as a novel and attractive gene delivery vehicle for mammalian cells. Porcine circovirus type 2 (PCV2) is known to be associated with post-weaning multisystemic wasting syndrome (PMWS), an emerging swine disease which results in tremendous economic losses. In this study, baculovirus pseudotyped with vesicular stomatitis virus glycoprotein (VSV-G) was used as a vector to express capsid (Cap) protein, the most important immunogen of PCV2, under the transcriptional control of cytomegalovirus immediate early (CMV-IE) enhancer/promoter. The resultant recombinant baculovirus (BV-G-ORF2) efficiently transduced and expressed the Cap protein in mammalian cells, as demonstrated by Western blot and flow cytometric analyses. After direct vaccination with 1x10(8) or 1x10(9)plaque forming units (PFU)/mouse of BV-G-ORF2, significant PCV2 specific ELISA antibodies, neutralizing antibodies, as well as cellular immune responses could be induced in mice. BV-G-ORF2 exhibited better immunogenicity than a DNA vaccine encoding the Cap protein, even at a dose of 1x10(8)PFU/mouse. Taken together, the improved immunogenicity of BV-G-ORF2, together with the unique advantages of pseudotype baculovirus, including easy manipulation, simple scale-up, lack of toxicity, and no pre-existing antibody against baculovirus in the hosts, indicate that pseudotype baculovirus-mediated gene delivery can be utilized as an alternative strategy to develop a new generation of vaccines against PCV2 infection. PMID- 18394723 TI - Novel roles for Nogo receptor in inflammation and disease. AB - The Nogo receptor (NgR), which was identified as a common receptor for three axon growth inhibitors associated with myelin, has been extensively characterized for its role in triggering growth cone collapse and arresting neurite/axon growth. Recent studies indicate that NgR is also expressed in nonneuronal cells and modulates macrophage responses during inflammation after peripheral nerve injury. In this article, we discuss the possibility that NgR might have wider effects on inflammation in a variety of neurological conditions ranging from central nervous system trauma to diseases such as multiple sclerosis or Alzheimer's disease. PMID- 18394724 TI - Synthesis of 1-beta-D-ribofuranosyl-3-ethynyl-[1,2,4]triazole and its in vitro and in vivo efficacy against Hantavirus. AB - There are no FDA approved drugs for the treatment of hemorrhagic fever with renal syndrome (HFRS), a serious human illnesses caused by hantaviruses. Clinical studies using ribavirin (RBV) to treat HFRS patients suggest that it provides an improved prognosis when given early in the course of disease. Given the unique antiviral activity of RBV and the lack of other lead scaffolds, we prepared a diverse series of 3-substituted 1,2,4-triazole-beta-ribosides and identified one with antiviral activity, 1-beta-d-ribofuranosyl-3-ethynyl-[1,2,4]triazole (ETAR). ETAR showed an EC(50) value of 10 and 4.4 microM for Hantaan virus (HTNV) and Andes virus, respectively. ETAR had weak activity against Crimean Congo hemorrhagic fever virus, but had no activity against Rift Valley fever virus. Intraperitoneally delivered ETAR offered protection to suckling mice challenged with HTNV with a approximately 25% survival at 12.5 and 25mg/kg ETAR, and a MTD of 17.1+/-0.7 days. ETAR was phosphorylated in Vero E6 cells to its 5' triphosphate and reduced cellular GTP levels. In contrast to RBV, ETAR did not increase mutation frequency of the HTNV genome, which suggests it has a different mechanism of action than RBV. ETAR is an exciting and promising lead compound that will be elaborated in further synthetic investigations as a framework for the rational design of new antivirals for treatment of HFRS. PMID- 18394725 TI - Structure and sequence motifs of siRNA linked with in vitro down-regulation of morbillivirus gene expression. AB - The most challenging task in RNA interference is the design of active small interfering RNA (siRNA) sequences. Numerous strategies have been published to select siRNA. They have proved effective in some applications but have failed in many others. Nonetheless, all existing guidelines have been devised to select effective siRNAs targeting human or murine genes. They may not be appropriate to select functional sequences that target genes from other organisms like viruses. In this study, we have analyzed 62 siRNA duplexes of 19 bases targeting three genes of three morbilliviruses. In those duplexes, we have checked which features are associated with siRNA functionality. Our results suggest that the intramolecular secondary structure of the targeted mRNA contributes to siRNA efficiency. We also confirm that the presence of at least the sequence motifs U13, A or U19, as well as the absence of G13, cooperate to increase siRNA knockdown rates. Additionally, we observe that G11 is linked with siRNA efficacy. We believe that an algorithm based on these findings may help in the selection of functional siRNA sequences directed against viral genes. PMID- 18394726 TI - The role of 5-HT(1A) receptors in learning and memory. AB - The ascending serotonin (5-HT) neurons innervate the cerebral cortex, hippocampus, septum and amygdala, all representing brain regions associated with various domains of cognition. The 5-HT innervation is diffuse and extensively arborized with few synaptic contacts, which indicates that 5-HT can affect a large number of neurons in a paracrine mode. Serotonin signaling is mediated by 14 receptor subtypes with different functional and transductional properties. The 5-HT(1A) subtype is of particular interest, since it is one of the main mediators of the action of 5-HT. Moreover, the 5-HT(1A) receptor regulates the activity of 5-HT neurons via autoreceptors, and it regulates the function of several neurotransmitter systems via postsynaptic receptors (heteroreceptors). This review assesses the pharmacological and genetic evidence that implicates the 5 HT(1A) receptor in learning and memory. The 5-HT(1A) receptors are in the position to influence the activity of glutamatergic, cholinergic and possibly GABAergic neurons in the cerebral cortex, hippocampus and in the septohippocampal projection, thereby affecting declarative and non-declarative memory functions. Moreover, the 5-HT(1A) receptor regulates several transduction mechanisms such as kinases and immediate early genes implicated in memory formation. Based on studies in rodents the stimulation of 5-HT(1A) receptors generally produces learning impairments by interfering with memory-encoding mechanisms. In contrast, antagonists of 5-HT(1A) receptors facilitate certain types of memory by enhancing hippocampal/cortical cholinergic and/or glutamatergic neurotransmission. Some data also support a potential role for the 5-HT(1A) receptor in memory consolidation. Available results also implicate the 5-HT(1A) receptor in the retrieval of aversive or emotional memories, supporting an involvement in reconsolidation. The contribution of 5-HT(1A) receptors in cognitive impairments in various psychiatric disorders is still unclear. However, there is evidence that 5-HT(1A) receptors may play differential roles in normal brain function and in psychopathological states. Taken together, the evidence indicates that the 5 HT(1A) receptor is a target for novel therapeutic advances in several neuropsychiatric disorders characterized by various cognitive deficits. PMID- 18394727 TI - Trophic interactions between the toxic dinoflagellate Alexandrium catenella and Mytilus chilensis: feeding and digestive behaviour to long-term exposure. AB - Trophic interactions between Mytilus chilensis and the toxic dinoflagellate Alexandrium catenella were studied over a 21-day period to determine the absorption kinetics of biochemical components and the enzymatic activity of the digestive gland (i.e. amylase, cellulase complex, laminarinase and protease). For the duration of the experiment, logarithmic relationships were established for the assimilatory balance and carbohydrase activities (amylase, laminarinase and cellulase) of the mussels fed with a toxic diet. The study shows that M. chilensis was able to develop mechanisms which allow exploitation of the toxic microalgae as a food source, despite that its feeding and digestive processes were affected during the first days of contamination. PMID- 18394728 TI - Carotenoid depletion and ovarian dysfunction in common bully exposed to a New Zealand pulp mill effluent. AB - In a previous cumulative impacts assessments of fish populations along the Waikato River, New Zealand, reduced ovary pigmentation was observed in the common bully (Gobiomorphus cotidianus) with an associated reduction in recruitment in the pulp mill-impacted Kopakorahi Arm of Lake Maraetai compared to an upstream reference site. Carotenoid pigments are typically responsible for the yellow to red colouration of fish eggs and a number of functions for these pigments such as pro-vitamins and antioxidants have been proposed. The purpose of the current study was first to quantify the observed pigment reduction and then to establish possible consequences of carotenoid deficiency. Significantly lower levels of all measured carotenoids were observed in the ovaries of fish from below the mill outfall compared to the reference site. Using in vitro incubations of ovarian follicles, it was also found that these fish had a significantly reduced ability to produce the sex steroids testosterone and estradiol. This study demonstrates changes in ovarian carotenoids associated with pulp mill effluent exposure. Significantly reduced biosynthetic capacity of ovarian follicles demonstrates reproductive endocrine disruption potential associated with exposure to this particular effluent. These results may provide an important link between reproductive dysfunction, oxidative stress and food chain modifications in pulp mill-impacted systems. PMID- 18394729 TI - Presence of substance P and the neurokinin-1 receptor in tenocytes of the human Achilles tendon. AB - Nerve signal substances, such as the tachykinin substance P (SP), may be involved in the changes that occur in response to tendinopathy (tendinosis). It is previously known that the level of SP innervation within tendon tissue is limited, but results of experimental studies have suggested that SP may have stimulatory, angiogenetic and healing effects in injured tendons. Therefore, it would be of interest to know if there is a local SP-supply in tendon tissue. In the present study, the patterns of expression of SP and its preferred receptor, the neurokinin-1 receptor (NK-1 R), in normal and tendinosis human Achilles tendons were analyzed by use of both immunohistochemistry and in situ hybridization. We found that there was expression of SP mRNA in tenocytes, and that tenocytes showed expression of NK-1 R at protein as well as mRNA levels. The observations concerning both SP and NK-1 R were most evident for tenocytes in tendinosis tendons. Our findings suggest that SP is produced in tendinosis tendons, and furthermore that SP has marked effects on the tenocytes via the NK-1 R. It cannot be excluded that the SP effects are of importance concerning the processes of reorganization and healing that occur for tendon tissue in tendinosis. In conclusion, it appears as if SPergic autocrine/paracrine effects occur in tendon tissue during the processes of tendinosis, hitherto unknown effects for human tendons. PMID- 18394730 TI - Feelings of well being in elderly people: relationship to physical activity and physical function. AB - The aim of the research was to investigate in a sample of Spanish elderly whether measures of physical activity and physical function are related to feelings of well being, and whether level of dependence is a moderator in the relation of well being, physical activity and physical function. The sample was a cohort of 151 elderly people (89 women and 62 men, aged 60-98 years) from the North of Spain. Participants completed surveys including demographic characteristics, and measures of physical activity (Yale Physical Activity Survey, YPAS), instrumental activities of daily living (Barthel Index, BI) and well being (Psychological Well Being Scale, from Spanish: Escala de Bienestar Psicologico=EBP). Components of the physical function were measured by the Senior Fitness Test (SFT). Upper and lower body strength, dynamic balance, aerobic endurance, self-reported weekly energy expenditure and physical activity total time were significantly correlated with both Material and Subjective well being. All components of physical function were significantly impaired in dependent subjects when compared to independent individuals of the same sex and physical activity category. Significant differences were also observed in Subjective well being among less active dependent or independent individuals. In conclusion, physical function and physical activity are related to feelings of well being, and results emphasize the positive functional and psychological effects of physical activity in dependent subjects. PMID- 18394731 TI - Efficacy and safety of drug-eluting stents in ST-segment elevation myocardial infarction: a meta-analysis of randomized trials. AB - BACKGROUND: Recent concerns have emerged on the potential higher risk of stent thrombosis after DES implantation, that might be even more pronounced among STEMI patients. Thus, the aim of the current study was to perform a meta-analysis to evaluate the benefits and safety of DES as compared to BMS in patients undergoing primary angioplasty for STEMI. METHODS: The literature was scanned by formal searches of electronic databases (MEDLINE and CENTRAL). We examined all completed randomized trials of DES for STEMI. The following key words were used for study selection: randomized trial, myocardial infarction, reperfusion, primary angioplasty, stenting, DES, sirolimus-eluting stent (SES), Cypher, paclitaxel eluting stent (PES), Taxus. Information on study design, type of stent, inclusion and exclusion criteria, primary endpoint, number of patients, angiographic and clinical outcome, were extracted by two investigators. Disagreements were resolved by consensus. RESULTS: A total of 11 trials were included in the meta analysis, involving 3605 patients (1888 or 52.3% randomized to DES and 1719 or 47.7% randomized to BMS). At 12 months follow-up, no significant difference was observed in mortality (4.1% vs 4.4%, OR [95% CI]=0.91 [0.66-1.27], p=0.59, reinfarction (3.1% vs 3.4%, OR [95% CI]=0.85 [0.58, 1.23], p=0.38 or stent thrombosis (1.6% vs 2.2%, OR [95% CI]=0.76 [0.47, 1.23], p=0.22), whereas DES were associated with a significant reduction in TVR (5.0% vs 12.6%, OR [95% CI]=0.36 [0.28, 0.47], p<0.0001). Safety and efficacy of DES were confirmed at 18 to 24 months follow-up (data available from 4 trials including 1178 patients). CONCLUSIONS: This meta-analysis shows that among selected STEMI patients undergoing primary angioplasty, SES and PES, as compared to BMS, are safe and associated with a significant reduction in TVR at 1 and 2 years follow-up. PMID- 18394732 TI - "Inverted Tako-Tsubo": transient apical-sparing cardiomyopathy. AB - We present a case of "inverted Tako-Tsubo" syndrome in a pre-menopausal woman with normal coronary arteries. Transient basal myocardial akinesia and ECG findings of healed myocardial infarction appeared shortly after emotional stress and resolved completely 4 days later. PMID- 18394733 TI - Quantitative epidemiology: progress and challenges. AB - This manuscript is derived from a presentation at the 2006 AVEPM--Schwabe Symposium which honoured the 2006 recipient of the Calvin Schwabe Award--Dr. S. Wayne Martin. Throughout his career, Dr. Martin was instrumental in furthering the development of quantitative epidemiology. This manuscript highlights some of the recent advances in quantitative methods used in veterinary epidemiology and identifies some of the challenges we still face in the use of these methods. Areas discussed are: methods for analyzing hierarchical data, Bayesian methods, survival analysis and integrating causal thinking and statistical analyses. PMID- 18394734 TI - Using magnetoencephalography in assessing auditory skills in infants and children. AB - In this paper, the use of magnetoencephalography (MEG) in studying the basic auditory skills in infants and children is reviewed. The auditory skills are related to perceiving sound onsets and offsets of sounds, extracting rules and regularities in sound environments, perceiving differences and changes in sounds, categorizing sound elements, allocating attention towards certain sounds or sound streams, and attaching semantic information into sounds. Studying each of these auditory skills with MEG in particular stimulation paradigms is shortly reviewed, including two examples of data sets in children. PMID- 18394735 TI - Multi-limb coordination and rhythmic variability under varying sensory availability conditions in children with DCD. AB - Children with Developmental Coordination Disorder (DCD) have sensory processing deficits; how do these influence the interface between sensory input and motor performance? Previously, we found that children with DCD were less able to organize and maintain a gross motor coordination task in time to an auditory cue, particularly at higher frequencies [Whitall, J., Getchell, N., McMenamin, S., Horn, C., Wilms-Floet, A., & Clark, J. (2006). Perception-action coupling in children with and without DCD: Frequency locking between task relevant auditory signals and motor responses in a dual motor task. Child: Care, Health, and Development, 32, 679-692]. In the present study, we examine the same task (clapping in-phase to marching on a platform) under conditions involving the removal of vision and hearing. Eleven children with DCD (mean=7.21, SD=0.52 years), 7 typically developing (TD) children (mean=6.95+/-0.72 years), and 10 adults performed continuous clapping while marching under four conditions: with vision and hearing, without vision, without hearing, and without both. Results showed no significant condition effects for any measure taken. The DCD group was more variable in phasing their claps and footfalls than both the adult group and the TD group. There were also significant group effects for inter-clap interval coefficient of variation and inter-footfall interval coefficient of variation, with the DCD group being the most variable for both measures. Coherence analysis between limb combinations (e.g., left arm-right arm, right arm-left leg) revealed that the adults exhibited significantly greater coherence for each combination than both of the children's groups. The TD group showed significantly greater coherence than the DCD group for every limb combination except foot-foot and left hand-right foot. Measures of approximate entropy indicated that adults differed from children both with and without DCD in the structure of the variability across a trial with adults showing more complexity. Children with DCD are able to accomplish a self-initiated gross-motor coordination task but with increased variability for most but not all measures compared to typically developing children. The availability of visual and/or auditory information does not play a significant role in stabilizing temporal coordination of this task, suggesting that these are not salient sources of information for this particular task. PMID- 18394736 TI - Increased associated movements: influence of attention deficits and movement difficulties. AB - This study was designed to examine whether increased associated movements (AMs) reflect motor difficulties or the symptoms associated with attention disorders. Four groups of male children (N=51) aged 6-8 years participated: Group 1 consisted of 13 children diagnosed with developmental coordination disorder (DCD); Group 2 consisted of 13 children diagnosed with attention deficit hyperactivity disorder (ADHD); Group 3 consisted of 10 children diagnosed with co occurring DCD and ADHD, and Group 4 was a control sample of 15 children, with no known movement or attention difficulties. Various AM tasks were selected from established assessments and previous research to measure AM severity. The results supported the hypothesis that increased severity of AMs reflect movement difficulties with children in the DCD and DCD/ADHD groups displaying significantly more AMs than children in the ADHD and control groups (p<.001). No differences were found between the ADHD only and control groups (p=.67) or the DCD and DCD/ADHD groups (p=.81) suggesting that AM severity is not influenced by the neurodevelopmental symptoms associated with ADHD. PMID- 18394737 TI - Processing plant persistent strains of Listeria monocytogenes appear to have a lower virulence potential than clinical strains in selected virulence models. AB - Listeria monocytogenes is an important foodborne bacterial pathogen that can colonize food processing equipment. One group of genetically similar L. monocytogenes strains (RAPD type 9) was recently shown to reside in several independent fish processing plants. Persistent strains are likely to contaminate food products, and it is important to determine their virulence potential to evaluate risk to consumers. We compared the behaviour of food processing persistent and clinical L. monocytogenes strains in four virulence models: Adhesion, invasion and intracellular growth was studied in an epithelial cell line, Caco-2; time to death in a nematode model, Caenorhabditis elegans and in a fruit fly model, Drosophila melanogaster and fecal shedding in a guinea pig model. All strains adhered to and grew in Caco-2 cells in similar levels. When exposed to 10(6) CFU/ml, two strains representing the persistent RAPD type 9 invaded Caco-2 cells in lower numbers (10(2)-10(3) CFU/ml) as compared to the four other strains (10(4)-10(6) CFU/ml), including food and human clinical strains. In the D. melanogaster model, the two RAPD type 9 strains were among the slowest to kill. Similarly, the time to reach 50% killed C. elegans worms was longer (110 h) for the RAPD type 9 strains than for the other four strains (80 h). The Scott A strain and one RAPD type 9 strain were suspended in whipping cream before being fed to guinea pigs and the persistent RAPD type 9 strain was isolated from feces in a lower level (approximately 10(2) CFU/g) than the Scott A strain (approximately 10(5) CFU/g) (P<0.05). The addition of NaCl has been shown to cause autoaggregation and increases adhesion of L. monocytogenes to plastic. However, growth in the presence of NaCl did not alter the behaviour of the tested L. monocytogenes strains in the virulence models. Overall, the two strains representing a very common fish processing plant persistent group (RAPD type 9) appear to have a lower virulence potential in all four virulence models than Scott A and a strain isolated from a clinical case of listeriosis. PMID- 18394738 TI - Low-coverage massively parallel pyrosequencing of cDNAs enables proteomics in non model species: comparison of a species-specific database generated by pyrosequencing with databases from related species for proteome analysis of pea chloroplast envelopes. AB - Proteomics is a valuable tool for establishing and comparing the protein content of defined tissues, cell types, or subcellular structures. Its use in non-model species is currently limited because the identification of peptides critically depends on sequence databases. In this study, we explored the potential of a preliminary cDNA database for the non-model species Pisum sativum created by a small number of massively parallel pyrosequencing (MPSS) runs for its use in proteomics and compared it to comprehensive cDNA databases from Medicago truncatula and Arabidopsis thaliana created by Sanger sequencing. Each database was used to identify proteins from a pea leaf chloroplast envelope preparation. It is shown that the pea database identified more proteins with higher accuracy, although the sequence quality was low and the sequence contigs were short compared to databases from model species. Although the number of identified proteins in non-species-specific databases could potentially be increased by lowering the threshold for successful protein identifications, this strategy markedly increases the number of wrongly identified proteins. The identification rate with non-species-specific databases correlated with spectral abundance but not with the predicted membrane helix content, and strong conservation is necessary but not sufficient for protein identification with a non-species specific database. It is concluded that massively parallel sequencing of cDNAs substantially increases the power of proteomics in non-model species. PMID- 18394739 TI - Chromatographic separation and kinetic properties of fructosyltransferase from Aureobasidium pullulans. AB - Efficient chromatographic separation of fructosyltransferase from Aureobasidium pullulans was achieved on a preparative scale using a weak anion-exchanger Sepabeads FP-DA. The recovery yield was about 70% and the purification factor reached a value of 28. The molecular weight of the enzyme determined by size exclusion chromatography was 570,000. The enzyme exhibited both hydrolytic and transferase activity when the latter was higher in the whole concentration range and completely dominating at higher sucrose concentrations. It was found that sucrose was the only donor of fructosyl moiety used in the transfer reaction. The initial rate method was used for the investigation of the kinetics of the action of fructosyltransferase on sucrose in the concentration range 30-2,430 mM. The initial transfructosylation rate was well fitted with a linear function of the sucrose activity where the activity coefficient was an exponentially decreasing function of the sucrose concentration. PMID- 18394740 TI - Additional virulence genes in conjunction with efficient selection scheme, and compatible culture regime enhance recovery of stable transgenic plants in cowpea via Agrobacterium tumefaciens-mediated transformation. AB - A critical step in the development of robust Agrobacterium tumefaciens-mediated transformation system in recalcitrant grain legume, cowpea is the establishment of optimal conditions for efficient T-DNA delivery into target tissue and recovery of transgenic plants. A dramatic increase in efficiency of T-DNA delivery was achieved by constitutive expression of additional vir genes in resident pSB1 vector in Agrobacterium strain LBA4404. A geneticin based selection system permitted rapid and efficient identification of transgenic shoots without interfering with their regeneration, and eliminated the bulk of escapes. Supplementation of 0.5 microM kinetin to medium containing 5.0 microM benzyl aminopurine after 1 week of culture followed by 3 weeks of culture were found critical for optimal multiplication and elongation of transformed shoots from cotyledonary node explants. Combining these three developments, we recovered fertile transgenic plants at a frequency of 1.64%, significantly higher than previous reports. The presence, integration, expression and inheritance of transgenes were confirmed by molecular analysis. The protocol developed for cultivar Pusa Komal will facilitate the transfer of desirable traits into cowpea. PMID- 18394741 TI - Improved piercing of microneedle arrays in dermatomed human skin by an impact insertion method. AB - An electrical applicator was designed, which can pierce short microneedles into the skin with a predefined velocity. Three different shapes of microneedles were used, namely 300 mum assembled hollow metal microneedle arrays, 300 mum solid metal microneedle arrays and 245 mum hollow silicon microneedle arrays. The latter are available as 4x4, 6x6 and 9x9 arrays. When using a velocity of 1 or 3 m/s reproducible piercing of dermatomed and full thickness human skin was evident from the appearance of blue spots on the dermal side of the skin after Trypan Blue treatment and the presence of fluorescently labeled particles in dermatomed skin. Manual piercing did not result in the appearance of blue spots. Transport studies revealed that i) piercing of microneedles with a predefined velocity into human skin resulted in a drastic enhancement of the Cascade Blue (CB, Mw 538) transport, ii) A higher piercing velocity resulted in a higher CB transport rate, iii) The CB transport rate was also dependent on the shape of the microneedles and iv) no difference in transport rate was observed between 4x4, 6x6 and 9x9 hollow silicon microneedle arrays. PMID- 18394742 TI - [Does exercise therapy for chronic lower-back pain require daily isokinetic reinforcement of the trunk muscles?]. AB - OBJECTIVE: The goal of this study was to determine the benefits of a functional retraining programme (with or without daily isokinetic reinforcement of the trunk muscles) in patients with lower-back pain. METHOD: Two groups of 30 patients took part in the study. The control group (CG) underwent a four-week reconditioning program in a day hospital, whereas a second interventional group (IG) additionally performed daily isokinetic training of the trunk muscles. Three evaluations were carried out: before hospitalization (T1), immediately after hospitalization (T2) and three months postrehabilitation (T3). RESULTS: We observed an improvement in each parameter after rehabilitation, regardless of the group. A decrease in the DALLAS scores revealed a reduced impact of lower-back pain on the patients' lives. Pain experienced fell by 24%, analgesic treatment was significantly decreased (CG: -53%; IG: -56%), muscle endurance was improved (quadriceps: +30%, abdominal muscles: +20%, paraspinal muscles: +23%, quadratus lumborum: +33%) and the patients were more supple, as revealed by a decrease in the finger-to-ground distance (at T1, CG: 12.9+/-6.1cm; IG: 13.6+/-5.5 cm at T1; CG: 2.2+/-5.4 cm; IG: 2.8+/-5.1cm at T2). The sole difference for CG and IG at T2 resulted from an improvement in the performance of the trunk extensor muscles, which was significantly greater in the IG (CG: +14%; IG: +20%). Three months after rehabilitation, the benefits were still present for the two groups and, indeed, were even greater for certain parameters. CONCLUSION: Regardless of the protocol, the patients improved in both physical and psychological terms and these improvements were maintained over a short period, at least. Our results confirmed that one functional recovery programme is not superior to another for patients with lower-back pain. PMID- 18394743 TI - Rosiglitazone improves insulin sensitivity with increased serum leptin levels in patients with type 2 diabetes mellitus. AB - Rosiglitazone (RSG) is known to be an agonist for the peroxisome proliferator activated receptor-gamma (PPARgamma) and promotes differentiation of pre adipocytes into adipocytes. Leptin is highly correlated with adiposity, while the activation of PPARgamma is known to inhibit Lep gene expression and leptin release. This study was performed to evaluate the relationship between changes in circulating leptin levels, insulin sensitivity and regional adiposity after RSG treatment. Two hundred fifty-one type 2 diabetic patients (176 men and 75 women) who had been treated with sulfonylurea and/or metformin received 4 mg of RSG daily, in addition to the previous medications. Before and after RSG treatment (average duration 5.6+/-0.9 months), indices of insulin resistance, metabolic parameters, and serum leptin and adiponectin levels were measured. Abdominal subcutaneous fat thickness (SFT(max)) and visceral fat thickness were measured by sonography. After RSG treatment, HOMA-IR index decreased significantly (2.82+/ 1.94 vs. 2.01+/-1.58), while BMI and SFT(max) increased, and leptin (4.72+/-3.77 vs. 5.69+/-4.30 ng/ml) and adiponectin levels (7.54+/-10.20 vs. 12.89+/-10.13 microg/ml) increased. The increase in serum leptin correlated with an increase in SFT(max) (r=0.511, p<0.001) and with a reduction in HOMA-IR (r=-0.368, p<0.001). The correlation of Delta leptin with Delta HOMA-IR and with Delta SFT(max) was higher in females and among insulin-resistant subjects. In conclusion, RSG improves the insulin sensitivity with increased serum leptin levels in patients with type 2 diabetes mellitus, which is related to an increase in subcutaneous adiposity. PMID- 18394744 TI - Antibody-targeted myofibroblast apoptosis reduces fibrosis during sustained liver injury. AB - BACKGROUND/AIMS: Myofibroblast apoptosis promotes the resolution of liver fibrosis. However, retaining macrophages may enhance reversal. The effects of specifically stimulating myofibroblast apoptosis in vivo were assessed. METHODS: A single chain antibody (C1-3) to an extracellular domain of a myofibroblast membrane protein was injected as a fluorescent- or gliotoxin conjugate into mice with liver fibrosis. RESULTS: C1-3 specifically targeted alpha-smooth muscle actin positive liver myofibroblasts within scar regions of the liver in vivo and did not co-localise with liver monocytes/macrophages. Injection of free gliotoxin stimulated a 2-fold increase in non-parenchymal cell apoptosis and depleted liver myofibroblasts by 30% and monocytes/macrophages by 50% but had no effect on fibrosis severity in the sustained injury model employed. In contrast, C1-3 targeted gliotoxin stimulated a 5-fold increase in non-parenchymal cell apoptosis, depleted liver myofibroblasts by 60%, did not affect the number of monocytes/macrophages and significantly reduced fibrosis severity. Fibrosis reduction was associated with increased metalloproteinase-13 levels. CONCLUSIONS: These data demonstrate that specific targeting of liver myofibroblast apoptosis is the most effective anti-fibrogenic therapy, supporting a role for liver monocytes and/or macrophages in the promotion of liver fibrosis reduction. PMID- 18394745 TI - Cost of hospital-wide activities to improve patient safety and infection control: a multi-centre study in Japan. AB - OBJECTIVE: The aim of this study was to assess the financial costs to hospitals for the implementation of hospital-wide patient safety and infection control programs. METHODS: We conducted questionnaire surveys and structured interviews in seven acute-care teaching hospitals with an established reputation for their efforts towards improving patient safety. We defined the scope of patient safety activities by use of an incremental activity measure between 1999 and 2004. Hospital-wide incremental manpower, material, and financial resources to implement patient safety programs were measured. RESULTS: The total incremental activities were 19,414-78,540 person-hours per year. The estimated incremental costs of activities for patient safety and infection control were calculated as US$ 1.100-2.335 million per year, equivalent to the employment of 17-40 full-time healthcare staff. The ratio of estimated costs to total medical revenue ranged from 0.55% to 2.57%. Smaller hospitals tend to shoulder a higher burden compared to larger hospitals. CONCLUSIONS: Our study provides a framework for measuring hospital-wide activities for patient safety. Study findings suggest that the total amount of resources is so great that cost-effective and evidence-based health policy is needed to assure the sustainability of hospital safety programs. PMID- 18394746 TI - Widespread and subtle: alternative splicing at short-distance tandem sites. AB - Alternative splicing at donor or acceptor sites located just a few nucleotides apart is widespread in many species. It results in subtle changes in the transcripts and often in the encoded proteins. Several of these tandem splice events contribute to the repertoire of functionally different proteins, whereas many are neutral or deleterious. Remarkably, some of the functional events are differentially spliced in tissues or developmental stages, whereas others exhibit constant splicing ratios, indicating that function is not always associated with differential splicing. Stochastic splice site selection seems to play a major role in these processes. Here, we review recent progress in understanding functional and evolutionary aspects as well as the mechanism of splicing at short distance tandem sites. PMID- 18394747 TI - Megakaryocyte potentiating factor as a tumor marker of malignant pleural mesothelioma: evaluation in comparison with mesothelin. AB - PURPOSE: An early and reliable blood test is one deficiency in diagnosis of malignant pleural mesothelioma (MPM). Megakaryocyte potentiating factor (MPF) and mesothelin variants (MSLN), members of the mesothelin gene family, have been studied as candidate serum markers for MPM. We developed a novel enzyme-linked immunosorbent assay (ELISA) system to compare the diagnostic efficacy of MPF and MSLN in MPM and control groups. EXPERIMENTAL DESIGN: MPF and MSLN were assayed with ELISA in 27 consecutive MPM patients and 129 controls including patients with lung cancer and asymptomatic asbestos-exposed subjects. RESULTS: Statistically significant elevation of serum MPF and MSLN levels was noted in MPM patients in comparison with every control group. The area under the receiver operating characteristic curve (AUC) was calculated for differentiation of MPM and lung cancer, healthy asbestos-exposed subjects, and healthy adults. While the AUC for serum MPF was 0.879, cut-off=19.1ng/ml (sensitivity=74.1%, specificity=90.4%), the AUC for serum MSLN was 0.713, cut-off=93.5ng/ml (sensitivity=59.3%, specificity=86.2%). Comparison between AUC for MPF and MSLN values shows that MPF is significantly superior to MSLN (p=0.025). Finally, there was a significant correlation between MPF and MSLN values for MPM (Pearson's correlation coefficient=0.77; p<0.001). CONCLUSIONS: These findings suggest that diagnostic value of MPF for MPM was better than that of MSLN although both markers showed almost equal specificity for MPM. PMID- 18394748 TI - Adenocarcinoma of lung in never smoked children. AB - Except in developed countries, the incidence of lung cancer notably in women and non-smokers is rising in most parts of the world. Here, we report two children diagnosed with pulmonary adenocarcinoma at a very early age. Interestingly, both showed negative EGFR mutation despite their ethnicity, histology, never smoking status and early age. Furthermore, one had preceding pulmonary tuberculosis. In the literature, the possible association of pulmonary tuberculosis and adenocarcinoma of lung especially in non-smokers has long been debated. The two children, by far the youngest with EGFR negative adenocarcinoma of lung, form the basis of this report. PMID- 18394749 TI - Differences in gene expression levels between early and later stages of human lung development are opposite to those between normal lung tissue and non-small lung cell carcinoma. AB - We, for the first time, directly compared gene expression profiles in human non small cell lung carcinomas (NSCLCs) and in human fetal lung development. Previously reported correlations of gene expression profiles between lung cancer and lung development, deduced from matching data on mouse development and human cancer, have brought important information, but suffered from different timing of mouse and human gene expression during fetal development and fundamental differences in tumorigenesis in mice and humans. We used the suppression subtractive hybridization technique to subtract cDNAs prepared from human fetal lung samples at weeks 10-12 and 22-24 and obtained a cDNA library enriched in the transcripts more abundant at the later stage. cDNAs sequencing and RT-PCR analysis of RNAs from human fetal and adult lungs revealed 12 differentially transcribed genes: ADH1B, AQP1, FOLR1, SLC34A2, CAV1, INMT, TXNIP, TPM4, ICAM-1, HLA-DRA, EFNA1 and HLA-E. Most of these genes were found up-regulated in mice and rats at later stages than in human lung development. In surgical samples of NSCLC, these genes were down-regulated as compared to surrounding normal tissues and normal lungs, thus demonstrating opposite expression profiles for the genes up-regulated during fetal lung development. PMID- 18394750 TI - Complete metabolic tumour response, assessed by 18-fluorodeoxyglucose positron emission tomography (18FDG-PET), after induction chemotherapy predicts a favourable outcome in patients with locally advanced non-small cell lung cancer (NSCLC). AB - BACKGROUND: 18FDG-PET and multislice computerized axial tomography (CT) scan are used for diagnosis, staging and response evaluation in NSCLC patients. The correlation between the response assessment by both imaging techniques and survival was assessed in patients with unresectable stage III NSCLC treated with induction chemotherapy followed by consolidation radiotherapy. METHODS: Thirty one patients, enrolled in a phase II study evaluating the efficacy and toxicity of a novel triplet induction chemotherapy (paclitaxel, carboplatin and gemcitabine) (PACCAGE) before consolidation radiotherapy, were evaluated by CT and 18FDG-PET at baseline and after three cycles of chemotherapy. The correlation between CT and 18FDG-PET response and time to progression and overall survival was analyzed using the Kaplan-Meier estimates of survival and the log rank test. RESULTS: Ten patients with a complete response (CR) on 18FDG-PET had a significantly longer time to progression and overall survival than patients with a non-CR (median 19.9 months versus 9.8 months, p=0.026, and median >49 months versus 14.4 months, p=0.004, respectively). Twenty patients with a partial CT response (PR) had a significantly longer time to progression (median 15 months versus 9.4 months, p=0.001) than patients with a non-PR but the difference in overall survival only showed a trend (23.3 months versus 14.4 months, p=0.093). CONCLUSIONS: A CR on 18FDG-PET following induction chemotherapy for locally advanced, unresectable NSCLC seems to be a more powerful prognostic marker for survival compared to PR on CT. PMID- 18394751 TI - Prevalence of meticillin-resistant Staphylococcus aureus in expressed breast milk in a neonatal intensive care unit. PMID- 18394752 TI - Pertussis outbreak on a neonatal unit: identification of a healthcare worker as the likely source. AB - We describe the investigation and containment of an outbreak of pertussis on a neonatal unit. Bacterial culture, polymerase chain reaction (PCR) and serology were used to confirm suspected cases. Two infants with pertussis were identified and a nurse with prolonged cough was traced as the likely source. Control interventions included mass chemoprophylaxis of healthcare workers and patients and exclusion from work of healthcare workers with cough. The use of PCR allowed rapid assessment of the extent of the outbreak. This outbreak highlights the risk to hospitalised infants posed by circulation of Bordetella pertussis in young adults and illustrates the utility of PCR in rapidly assessing the extent of outbreaks. Prevention strategies such as universal vaccination of adolescents, or selective vaccination of healthcare workers, should be considered in the UK. PMID- 18394753 TI - Successful control of an outbreak of invasive aspergillosis in a regional haematology unit during hospital construction works. AB - We report the successful control of an outbreak of six cases of nosocomial invasive aspergillosis (IA) in our haematology unit coinciding with major hospital construction works. Infection control changes included unit relocation, impermeable barriers at construction site, face-masking and voriconazole prophylaxis of 18 further high-risk patients, none of which developed breakthrough IA. A multi-faceted pre-emptive approach involving clinicians, hospital management, engineering and building departments is essential in preventing nosocomial IA outbreaks. PMID- 18394754 TI - Are meticillin-resistant Staphylococcus aureus bloodstream infection targets fair to those with other types of healthcare-associated infection or cost-effective? AB - UK National Health Service Trust hospitals are required to report meticillin resistant Staphylococcus aureus bloodstream infection (MRSA BSI) and also have targets for their reduction. We argue that the MRSA BSI target influences local priorities, and that the high priority given to control of MRSA BSI is not fair or cost-effective when account is taken of the much larger number of patients who develop other types of serious hospital-acquired infection. Reasons for infection control priorities should be made explicit. Use of a formal and reasoned process of priority setting would also counteract the potential distortion of local priorities by national targets. PMID- 18394755 TI - Evidence for postsynaptic modulation of muscle contraction by a Drosophila neuropeptide. AB - DPKQDFMRFamide, the most abundant FMRFamide-like peptide in Drosophila melanogaster, has been shown previously to enhance contractions of larval body wall muscles elicited by nerve stimulation and to increase excitatory junction potentials (EJPs). The present work investigated the possibility that this peptide can also stimulate muscle contraction by a direct action on muscle fibers. DPKQDFMRFamide induced slow contractions and increased tonus in body wall muscles of Drosophila larvae from which the central nervous system had been removed. The threshold for this effect was approximately 10(-8)M. The increase in tonus persisted in the presence of 7x10(-3)M glutamate, which desensitized postsynaptic glutamate receptors. Thus, the effect on tonus could not be explained by enhanced release of glutamate from synaptic terminals and, thus, may represent a postsynaptic effect. The effect on tonus was abolished in calcium free saline and by treatment with L-type calcium channel blockers, nifedipine and nicardipine, but not by T-type blockers, amiloride and flunarizine. The present results provide evidence that this Drosophila peptide can act postsynaptically in addition to its apparent presynaptic effects, and that the postsynaptic effect requires influx through L-type calcium channels. PMID- 18394756 TI - IGF-1 signaling reduces neuro-inflammatory response and sensitivity of neurons to MPTP. AB - Reduced expression of IGF-1R increases lifespan and resistance to oxidative stress in the mouse, raising the possibility that this also confers relative protection against the pro-parkinsonian neurotoxin MPTP, known to involve an oxidative stress component. We used heterozygous IGF-1R(+/-) mice and challenged them with MPTP. Interestingly, MPTP induced more severe lesions of dopaminergic neurons of the substantia nigra, in IGF-1R(+/-) mice than in wild-type animals. Using electron spin resonance, we found that free radicals were decreased in IGF 1R(+/-) mice in comparison with controls, both before and after MPTP exposure, suggesting that the increased vulnerability of dopamine neurons is not caused by oxidative stress. Importantly, we showed that IGF-1R(+/-) mice display a dramatically increased neuro-inflammatory response to MPTP that may ground the observed increase in neuronal death. Microarray analysis revealed that oxidative stress-associated genes, but also several anti-inflammatory signaling pathways were downregulated under control conditions in IGF-1R(+/-) mice compared to WT. Collectively, these data indicate that IGF signaling can reduce neuro inflammation dependent sensitivity of neurons to MPTP. PMID- 18394757 TI - Ultrastructures and strain comparison of under-glycosylated scrapie prion fibrils. AB - Prions, composed primarily of misfolded, often fibrillar, polymers of prion protein, have poorly understood structures. Heavy surface glycosylation may obscure visualization of their fibrillar cores, so we purified severely under glycosylated prion protein fibrils from scrapie-infected transgenic mice expressing anchorless prion protein. Using electron and atomic force microscopy, we obtained dimensions and morphological information about prion protein core protofilaments which variably intertwined to form scrapie fibrils. Occasional isolated protofilaments were observed, suggesting that the lateral association of protofilaments is neither essential nor invariant in prion protein polymerization. Strain comparisons suggested basic structural differences; ME7 and 22L fibrils contained thinner protofilaments, 22L fibrils preferred left handed twists, and 22L fibril periodicities averaged 106nm per half-turn, compared with 64 and 66nm for RML and ME7 fibrils, respectively. The strains displayed overlapping fibril morphologies, providing evidence that prion fibril morphology is influenced, but not dictated, by strain-dependent differences in protofilament structure. These measurements of the amyloid core of scrapie fibrils should aid development of models of prion structure and strain determination. PMID- 18394758 TI - Synthesis and anti-HIV-1 integrase activities of 3-aroyl-2,3-dihydro-1,1-dioxo 1,4,2-benzodithiazines. AB - A series of novel 3-aroyl-2,3-dihydro-1,1-dioxo-1,4,2-benzodithiazines 15-28 as potential HIV-1 integrase (IN) inhibitors have been synthesized by the reduction of 3-aroyl-1,1-dioxo-1,4,2-benzodithiazines 1-14 with benzenesulfonyl hydrazide. All the compounds 15-28 inhibited IN mediated strand transfer reaction with IC(50) values ranging from 3 to 30 microM. The 3-(4-bromobenzoyl)-6-chloro-7 methyl-2,3-dihydro-1,1-dioxo-1,4,2-benzodithiazine 17 with the IC(50) values of 4+/-1 and 3+/-1 microM for 3'-processing and strand transfer, respectively, was the most potent. Compound 17 as well its analogues were 5-20-fold less potent in Y99S and H114A mutants, implicating these residues as potential drug-binding site. This is a first report implicating Y99S and H114A of IN core domain in drug binding interactions. PMID- 18394759 TI - [Rare thrombophilic states]. AB - PURPOSE: Rare thrombophilic states are mostly associated with recurrent venous thrombosis or severe thrombosis such as neonatal purpura fulminans. We review here the various causes of rare thrombophilic states. CURRENT KNOWLEDGE AND KEY POINTS: Rare thrombophilic states associated with recurrent venous thrombosis include the following: antithrombin deficiencies, homozygous for protein C or protein S deficiency or compound heterozygous, double heterozygous for genetic thrombophilia and the rare thrombophilia due to auto-antibodies. Their frequency in patients with venous thromboembolism is below 2%. FUTURE PROSPECTS AND PROJECTS: These uncommon thrombophilic states require a treatment in a specialized department. PMID- 18394760 TI - [XIth International myeloma workshop, 2007, June 25-30, Kos island, Greece]. PMID- 18394761 TI - [C-reactive protein (CRP) after revascularized STEMI: is CRP a prognostic factor?]. AB - PURPOSE: During myocardial infarction (MI), numerous biomarkers increase, such as troponin (necrosis), BNP, and high sensibility C-reactive protein (hsCRP) (inflammation). The objectives of the study were to study kinetics of hsCRP after a revascularized MI, and correlations between hsCRP and clinical outcomes or biological markers, and prognostic value of CRP. PATIENTS AND METHODS: Fifty-two patients were admitted for STEMI (ST segment Elevation MI). Primary coronarography interventions (PCI) were performed for urgent reperfusion. Patients were included only in case of success (TIMI 3). Clinical examination was completed by a biological follow-up of BNP, troponin-I (before and after PCI, days 1, 2, 3, 6) and hsCRP (days 0, 1, 2, 3, 6). Clinical outcomes follow-up was performed during hospitalization, on the first month, and the sixth month. RESULTS: hsCRP increases during the first days (peak on day 3: 46.1mg/L), and decreases between the third and the seventh day. Clinical outcomes were correlated with CRP: door-to-balloon time, age, creatinin level on admission. During follow-up, there were clinical events in 13/49 (26%) of the patients. Among them, hsCRP on day 2 was higher (p < 0.0001), compared to other patients. Compared to other biological markers, hsCRP was correlated with BNP on days 2 and 3 (p = 0.008). CONCLUSION: hsCRP increases after revascularized STEMI, in accordance to the infarct size, in the first days. hsCRP is correlated with cardiovascular pronostic biomarkers. hsCRP could play an active role, and could be used as a pronostic biomarker after revascularized STEMI, which are usually considered as a low-risk population. PMID- 18394762 TI - Promoting genuine consumer participation in mental health education: a consumer academic role. AB - Mental health policy reform in Australia has led to the expectation of increased consumer participation not only in all aspects of service delivery, but also in the education and training of mental health professionals. The aim of this paper is to explore the development and introduction of a role for a consumer of mental health services within an academic institution, including achievements of the role and the principles contributing to successful implementation. The consumer academic position was successfully implemented and has proved a valuable role. The success of the role was considered to be dependent on the following factors: partnership and commitment, support, scope and autonomy. In conclusion, a consumer can play a valuable role within an academic institution. However, success will depend upon a number of important environmental factors that promote the independence of the consumer voice. PMID- 18394763 TI - Increasing the humoral immunogenic properties of the HIV-1 Tat protein using a ligand-stabilizing strategy. AB - Tat is regarded as an attractive target for the development of an AIDS vaccine. However, works suggest that Tat is a poorly immunogenic protein and therefore we attempted to increase its immunogenic potency. As we observed that Tat is highly sensitive to enzymatic degradation in vitro we tried to make it less susceptible to proteolysis using ligands. We complexed Tat101 with various sulfated sugars and observed that some of these ligands made the protein more resistant to proteolysis and more immunogenic. In a more thorough study, we observed that a low-molecular-weight heparin fragment, called Hep6000, altered both the cell binding capacity and transactivating activity of Tat101, suggesting that this sulfated polysaccharide can make the protein less toxic. Sera raised against Tat101 and Tat101/Hep6000 similarly bound mainly to the N-terminal region of the protein, indicating that formation of the complex does not alter the B-cell immunodominant region. Anti-Tat101/Hep6000 antisera neutralized the transactivating activity of Tat101 more efficiently than anti-Tat101 antisera. Altogether, these results indicate that stabilization of Tat101 using sulfated sugars increases its immunogenicity and might be of value in increasing its vaccine efficacy. PMID- 18394764 TI - Informing adolescents about human papillomavirus vaccination: what will parents allow? AB - With the introduction of human papillomavirus (HPV) vaccination an evidence base on effective adolescent educational interventions is urgently required. We undertook formative research to develop and evaluate a film on HPV and cervical cancer prevention for school children who will be offered HPV vaccination in the UK. The main outcome measures were the number of children allowed by parents to view the film and children's knowledge. Our results indicated that the film's four key messages were acceptable to parents and largely understood by adolescents but these messages will need reinforcing if the full potential of a prophylactic vaccine is to be realised. PMID- 18394765 TI - Comparison of the immunogenicity and safety of measles vaccine administered alone or with live, attenuated Japanese encephalitis SA 14-14-2 vaccine in Philippine infants. AB - Japanese encephalitis (JE) virus is a major cause of disease, disability, and death in Asia. An effective, live, attenuated JE vaccine (LJEV) is available; however, its use in routine immunization schedules is hampered by lack of data on concomitant administration with measles vaccine (MV). This study evaluated the immunogenicity and reactogenicity of LJEV and MV when administered at the same or separate study visits in infants younger than 1 year of age. Three groups of healthy infants were randomized to receive LJEV at age of 8 months and MV at 9 months (Group 1; n=100); MV and LJEV together at 9 months (Group 2; n=236); or MV and LJEV at 9 and 10 months, respectively (Group 3; n=235). Blood was obtained 4 weeks after each vaccine administration to determine antibody levels for measles and JE. Reactogenicity was assessed by parental diaries and clinic visits. Four weeks after immunization, measles seroprotection rates (defined as > or =340 mIU/ml) were high and comparable in all three groups and specifically, rates in the combined MV-LJEV (Group 2) were not statistically inferior to those in Group 3 receiving MV separately (96% versus 100%, respectively). Likewise, the LJEV seroprotection rates were high and similar between the three groups. The reactogenicity profiles of the three vaccine schedules were also analogous. LJEV and MV administered together are well tolerated and immunogenic in infants younger than 1 year. These results should facilitate incorporation of LJEV into routine immunization schedules with MV. PMID- 18394766 TI - The novice birthing: theorising first-time mothers' experiences of birth at home and in hospital in Australia. AB - OBJECTIVE: to explore first-time mothers' experiences of birth at home and in hospital in Australia. DESIGN: a grounded theory methodology was used. Data were generated from in-depth interviews with women in their own homes. SETTING: Sydney, Australia. PARTICIPANTS: 19 women were interviewed. Seven women who gave in a public hospital and seven women who gave birth for the first time at home were interviewed and their experiences were contrasted with two mothers who gave birth for the first time in a birth centre, one mother who gave birth for the first time in a private hospital and two women who had given birth more than once. FINDINGS: three categories emerged from the analysis: preparing for birth, the novice birthing and processing the birth. These women shared a common core experience of seeing that they gave birth as 'novices'. The basic social process running through their experience of birth, regardless of birth setting, was that, as novices, they were all 'reacting to the unknown'. The mediating factors that influenced the birth experiences of these first-time mothers were preparation, choice and control, information and communication, and support. The quality of midwifery care both facilitated and hindered these needs, contributing to the women's perceptions of being 'honoured'. The women who gave birth at home seemed to have more positive birth experiences. IMPLICATIONS FOR PRACTICE: identifying the novice status of first-time mothers and understanding the way in which they experience birth better explains previous research that reports unrealistic expectations and fear that may be associated with first-time birthing. It demonstrates how midwives can contribute to positive birth experiences by being aware that first-time mothers, irrespective of birth setting, are essentially reacting to the unknown as they negotiate the experience of birth. PMID- 18394767 TI - Rapporteur's Report Session 3: exposure to grunerite (amosite) asbestos: historical perspectives of the health effects. PMID- 18394768 TI - Pharmacotherapy of schizophrenia with comorbid substance use disorder--reviewing the evidence and clinical recommendations. AB - Substance use disorder is the most common psychiatric comorbidity in schizophrenic patients, with prevalence rates of up to 65%. Recommendations for antipsychotic pharmacotherapy in schizophrenia are based on studies that excluded patients with this dual diagnosis. In the present comprehensive systematic review, the pharmacological studies performed in this subgroup of patients are summarised and discussed from the standpoint of evidence-based medicine. Unfortunately, randomized controlled studies, providing a high evidence level, in patients with this dual diagnosis are rare. Data, mainly based on open studies or case series, suggest superior efficacy for second generation antipsychotic agents (SGAs) (aripiprazole, clozapine, olanzapine, quetiapine, risperidone) with regard to improvement of distinct psychopathological symptoms, reduced craving and greater reduction of substance use compared with orally administered conventional antipsychotics (FGAs). Tricyclic antidepressants given adjunctive to antipsychotic maintenance therapy showed efficacy in reducing substance use and craving. The administration of anti-craving agents (naltrexone) led to a decrease of drug intake. Unfortunately, there is no clinical experience with acamprosate in schizophrenic patients with comorbid alcoholism. In conclusion, there are more theoretically based arguments for the preferential use of SGAs in schizophrenic patients with comorbid substance use disorder while the empirical evidence is weak. The early initiation of treatment with antidepressants, depending on the patient's psychopathology, as well as add-on medication with anti-craving agents should be considered. PMID- 18394769 TI - Comparative study of pulmonary responses to nano- and submicron-sized ferric oxide in rats. AB - Ferric oxide (Fe(2)O(3)) nanoparticles are of considerable interest for application in nanotechnology related fields. However, as iron being a highly redox-active transition metal, the safety of iron nanomaterials need to be further studied. In this study, the size, dose and time dependent of Fe(2)O(3) nanoparticle on pulmonary and coagulation system have been studied after intratracheal instillation. The Fe(2)O(3) nanoparticles with mean diameters of 22 and 280 nm, respectively, were intratracheally instilled to male Sprague Dawley rats at low (0.8 mg/kgbw) and high (20 mg/kgbw) doses. The toxic effects were monitored in the post-instilled 1, 7 and 30 days. Our results showed that the Fe(2)O(3) nanoparticle exposure could induce oxidative stress in lung. Alveolar macrophage (AM) over-loading of phagocytosed nanoparticle by high dose treatment had occurred, while the non-phagocytosed particles were found entering into alveolar epithelial in day 1 after exposure. Several inflammatory reactions including inflammatory and immune cells increase, clinical pathological changes: follicular hyperplasia, protein effusion, pulmonary capillary vessel hyperaemia and alveolar lipoproteinosis in lung were observed. The sustain burden of particles in AM and epithelium cells has caused lung emphysema and pro-sign of lung fibrosis. At the post-instilled day 30, the typical coagulation parameters, prothrombin time (PT) and activated partial thromboplastin time (APTT) in blood of low dose 22 nm-Fe(2)O(3) treated rats were significantly longer than the controls. We concluded that both of the two-sized Fe(2)O(3) particle intratracheal exposure could induce lung injury. Comparing with the submicron sized Fe(2)O(3) particle, the nano-sized Fe(2)O(3) particle may increase microvascular permeability and cell lysis in lung epitheliums and disturb blood coagulation parameters significantly. PMID- 18394770 TI - Sex differences in denture satisfaction. AB - OBJECTIVES: Males and females differ in their responses to many health conditions and treatments. The aim of this randomized clinical trial was to determine whether there are differences in the way that male and female edentulous elders rate their satisfaction with new mandibular implant overdentures (IODs) and conventional dentures (CDs), at 6 and 12 months following delivery. METHODS: Edentulous elders (n=256) were randomly assigned to receive maxillary conventional dentures and either mandibular overdentures supported by two implants with ball attachments or conventional dentures. Participants rated their general satisfaction, as well as other features of their dentures (comfort, stability, ability to chew, aesthetics, etc.) prior to treatment and 6 and 12 months after delivery. RESULTS: Ratings of satisfaction with IODs were significantly higher than with CDs. Six months after delivery, females in the CD group rated their general satisfaction and satisfaction with ability to chew and aesthetics significantly lower than did the males. The sex differences in the CD group remained at 12 months after delivery. However, males and females in the IOD group rated their general satisfaction and all six subcategories equally. CONCLUSIONS: Elderly females are less satisfied with conventional dentures than elderly males with regards to aesthetics and ability to chew, but equally satisfied with implant overdentures. At 6 and 12 months after delivery, elderly edentulous males and females wearing mandibular implant overdentures were significantly more satisfied than those wearing conventional dentures. PMID- 18394771 TI - Artificial enamel dental caries treated with different topical fluoride regimes: an in situ study. AB - OBJECTIVES: To evaluate whether the topical fluoride application (acidulated phosphate fluoride, APF) at high concentration has an additional effect on the control of enamel lesions compared to fluoride dentifrice (FD; low concentration). The frequency of APF treatment on the arrestment of caries lesions and the amount of fluoride deposited on enamel after application of high and low fluoride concentrations were also evaluated. METHODS: Five subjects wore partial dentures with in vitro demineralized blocks during 35 days. All specimens (5p/subject) were brushed three times daily for 1 min with FD (1100 ppmF). Besides the FD treatment, four specimens were submitted to APF gel topical applications (12,300 ppmF) on weekly intervals (one to four applications). The enamel blocks were analyzed at baseline, after demineralization and after intraoral procedures regarding: surface roughness (SR), clinical aspects (brightness and texture), surface microhardness (SMH) and enamel fluoride content. Friedman's test was used to compare SMH and SR among the treatments. Analysis of variance, followed by Tukey's studentized range test, was used to evaluate fluoride content and SR among the groups. The significance level used was 5%. RESULTS: Changes in surface brightness, texture and SR were not detected. FD+3 APF and FD+4 APF were the only treatments capable of increasing SMH values and fluoride content compared to demineralized blocks (p < 0.001), although no differences could be observed within the treatments. CONCLUSIONS: This study showed that > or = 3 APF in addition to FD enhance enamel rehardening and produce a larger reservoir of fluoride. PMID- 18394772 TI - Nifedipine gastrointestinal therapeutic system (GITS) as an alternative to slow release for tocolysis--tolerance and pharmacokinetic profile. AB - OBJECTIVE: To determine nifedipine plasma concentrations after a loading dose of nifedipine 10mg capsules, 40 mg over 1h followed by slow-release tablets (60 mg/d) versus gastrointestinal therapeutic system (GITS) tablets (90 mg/d) for tocolysis. STUDY DESIGN: Prospective study in 14 pregnant women treated for threatened preterm labor. RESULTS: Following capsule administration there was a rapid rise in plasma concentration of drug achieving a peak of 97.5 microg/l (median) at 1h, then declined to 59.5 microg/l (median) at 5h. The concentration measured at 7200 min (120 h) was non-significantly higher in the slow-release group (median 25.5, range 6.9-67.2 microg/l) than in the GITS group (median 14.6, range 6.0-20.0 microg/l). Area under the curve (AUC) increased with the applied dose in both groups in a linear regression. Headache was more frequent in the slow-release group than in the GITS group (P=0.001). CONCLUSIONS: GITS tablets 90 mg/d are an alternative dosage regimen to previous used slow-release tablets 60 mg/d for tocolysis with similar pharmacokinetic profile and a good tolerance. However, tocolysis with GITS tablets is simpler than that with slow-release tablets and may be associated with a higher compliance. GITS tablets are therefore also qualified for home monitoring. PMID- 18394773 TI - Glucose metabolism and angiogenesis in granulosa cell tumors of the ovary: activation of Akt, expression of M2PK, TKTL1 and VEGF. AB - OBJECTIVES: Very little is known about the biology of granulosa cell tumors of the ovary. A hallmark of granulosa cell tumors of the ovary is extensive growth, distant metastases however, are rarely found. We hypothesise that granulosa cell tumors of the ovary on the one hand need to stimulate vascularisation; on the other hand glucose metabolism has to be altered to ensure the supply of nutrients and metabolites. Increased glycolysis, the main source of energy supply, is considered to be important during malignant transformation. Thus, we focussed on a selection of key factors in angiogenesis and tumor glycolysis to study metabolic characteristics of granulosa cell tumors of the ovary. STUDY DESIGN: We analysed 32 tumor specimens for immunohistochemical expression of vascular endothelial growth factor, phosphorylated Akt, M2 pyruvate kinase isoenzyme, and transketolase-like enzyme 1. As controls, we stained 10 samples of normal ovaries. RESULTS: We found expression of vascular endothelial growth factor in 94%, transketolase-like enzyme 1 in 81%, and phosphorylated Akt as well as M2 pyruvate kinase isoenzyme in 53% of the specimens. There were no significant differences between the expression levels in primary and those in recurrent tumors. Temporal analysis of marker expression in primary tumors and recurrences in the same patients revealed no increase or decrease of marker expression overtime. In contrast to granulosa cell tumors, normal ovaries showed no expression of the markers analysed in granulosa cells. CONCLUSION: Our results show that granulosa cell tumors of the ovary express vascular endothelial growth factor as an important stimulator of tumor angiogenesis as well as several molecular markers for glycolysis. The dependence of granulosa cell tumors of the ovary on the glycolytic pathway may provide a biochemical basis for therapeutic strategies involving glycolytic inhibitors. PMID- 18394774 TI - Factors determining detergent resistance of erythrocyte membranes. AB - The degree of detergent insolubility of cell membranes is a useful parameter to test the strength of lipid-lipid interactions relative to lipid-detergent interactions. Thus, solubility studies could give insights about lipid-lipid interactions relevant in domain formation. In this work we perform a detailed study of the solubilization of four different erythrocyte membrane systems: intact human and bovine erythrocytes, and human and bovine erythrocytes depleted in cholesterol with methyl-beta-cyclodextrin. Each system was incubated with different concentrations of the non-ionic detergent Triton X-100, and the insoluble fraction was characterized by determining cholesterol and phosphorus content. A distinct solubilization behavior was obtained for the four systems, which was quantified by a "detergent resistance parameter" obtained from the fit of the solubility curves. In order to correlate these findings with membrane structural parameters, we quantify the degree of acyl chain order/rigidity of the original membranes by EPR spectroscopy, finding that detergent resistance is higher when acyl chains are more rigid. Regarding compositional properties, we found a good correlation between detergent resistance parameters and the total amount of cholesterol plus sphingomyelin in the original membranes. Our results suggest that a high degree of acyl chain packing is the determinant membrane factor for resistance to the action of Triton X-100 in erythrocytes. PMID- 18394776 TI - Household behaviour and attitudes with respect to recycling food waste- experiences from focus groups. AB - It is a challenge to reduce the ever-increasing flow of waste. In Norway the systems for recycling of organic waste, paper, glass, metals, etc. differ between municipalities, both with regard to organizational and to technological structures. Our hypothesis is that people's attitudes and behaviour may differ with different systems of waste management. People's behaviour and attitudes regarding (organic) waste recycling were investigated in two municipalities with differing technical and organisational systems. Data came from interviews with municipal employees, questionnaires, focus groups and multi-criteria mapping. People seem to be better informed and more positive about organic waste recycling in one of the municipalities (MH, which has recycling of organic waste) than in the other (MS, which has no such recycling). The two municipalities had similar sets of important criteria for waste management (price, environmental friendliness, easy solutions, information). Many participants stated that they had learned from the group process, though only a few reported changed preferences. The institutional context seems to be important for people's behaviour and attitudes towards waste management. This implies that people's recycling behaviour does not only depend on technical and organisational aspects, but also on institutions. These are important messages for policy makers. On an individual basis, the different systems in the two municipalities seem to affect people's stated attitudes. These differences diminish when they are in a common setting where process and dialogue stimulate new thoughts and encourage people to act more altruistically. PMID- 18394775 TI - Tightly regulated gene expression in human hematopoietic stem cells after transduction with helper-dependent Ad5/35 vectors. AB - OBJECTIVE: Inducible and transient expression of transcription factors, growth factors, or mitogenic factors can be used to influence proliferation or differentiation of hematopoietic progenitor/stem cells (HSC). Furthermore, transient expression of proteins with site-specific endonuclease activity, potentially, can support targeted integration of exogenous transgenes into specific sites in the genome, a task that is currently a focus in development of gene therapy vectors. MATERIALS AND METHODS: We constructed a set of helper dependent adenovirus (Ad) vectors with serotype 35 fiber knob domains (HD Ad5/35), which can efficiently transduce human CD34(+) cells, particularly subsets with potential stem cell capacity. These vectors contain Tet-inducible expression systems that were shielded by insulators and transcription stop signals to minimize unspecific interference by transcriptional elements present in viral and stuffer DNA. We compared two vectors, containing a fusion between the Kruppel-associated box (KRAB) domain and the tetracycline repressor (HD Ad5/35.Tet-1) or an autoregulated rtTA (HD-Ad5/35.Tet-2) for regulated transgene expression in Mo7e cells, a model for HSC, and primary human CD34(+) cells. RESULTS: HD-Ad5/35.Tet-1 conferred lower background expression than HD-Ad5/35.Tet 2, although levels of induced gene expression were higher for HD-Ad5/35.Tet-2. In CD34(+) cells, while HD-Ad5/35.Tet-1 allowed for activated gene expression in all transduced cells, induced gene expression from HD-Ad5/35.Tet-2 was restricted to a small subset of CD34(+) cells. Importantly, clonogeneic assays and repopulation studies in nonobese diabetic/severe combined immunodeficient mice showed that both HD-Ad5/35.Tet-1 and -2 vectors mediated induced gene expression in primitive hematopoietic cells. These studies also confirmed that transduction of CD34(+) cells with HD-Ad5/35 vectors is not associated with cytotoxicity, a problem observed with first-generation Ad5/35 vectors. CONCLUSIONS: Both HD-Ad5/35 vector expression systems confer tightly regulated, transient transgene expression in human HSC. PMID- 18394777 TI - Extracorporeal shock wave therapy for management of chronic ulcers in the lower extremities. AB - Management of chronic ulcers in the lower extremities is still a challenge for patients and health providers. Recent studies showed extracorporeal shock waves (ESW) as effective in stimulating growth factors, inducing angiogenesis and healing of fractures and injuries. This study was planned to investigate the opportunity of introducing the ESW in the treatment of chronic wounds. Thirty consecutive patients with chronic posttraumatic, venous and diabetic ulcers, unresponsive to conservative or advanced dressing treatments, were counseled about the use of ESW as alternative treatment for their wounds. Thirty-two wounds were treated and 16 wounds healed completely within six sessions of ESW. In all of the nonhealed wounds, decrease of the amount of exudates, increased percentage of granulation tissue compared with fibrin/necrotic tissue and decrease of wounds' size were statistically significant after four to six sessions of ESW (p < 0.01). Significant decrease of pain was reported (p < 0.001). Comparison with a control group of 10 patients with chronic ulcer treated on the basis of regular dressings confirmed the statistical significant improvement in the healing process (p < 0.01). ESW therapy seems to be a safe, feasible and cost-effective treatment for chronic ulcers in the lower extremities. Further research and clinical trials are necessary to evaluate dose and time intervals of sessions to standardize a protocol of treatment in the management of chronic wounds. PMID- 18394779 TI - Editorial comment on: ejaculation elicited by microstimulation of lumbar spinothalamic neurons. PMID- 18394780 TI - Editorial comment on: training in percutaneous nephrolithotomy--a critical review. PMID- 18394781 TI - Editorial comment on: training in percutaneous nephrolithotomy--a critical review. PMID- 18394782 TI - Ejaculation elicited by microstimulation of lumbar spinothalamic neurons. AB - BACKGROUND: Neuroanatomical and lesion studies have identified lumbar spinothalamic (LSt) neurons to be essential for ejaculation, but their precise role remains elusive. OBJECTIVE: To assess the role of LSt neurons as a spinal pattern generator for ejaculation (SGE) and their action on anatomical structures involved in the two ejaculation phases, the emission and expulsion of semen. DESIGN: The bulbospongiosus muscle (BSM) was implanted with electrodes and the seminal vesicle (SV) or vas deferens (VD) lumen catheterized in adult anaesthetized rats. Spinal exposure at the fourth lumbar segment (L4) allowed lowering an electrode stereotaxically into area VII/X for brief (300-500ms) electrical stimulation of LSt neurons, while recording BSM-EMG and intraluminal SV or VD pressure. RESULTS: Brief electrical microstimulation in the LSt neuron area evoked the expulsion of semen in 17 of 17 rats, with motile spermatozoa in 10 of 17 rats. After stimulation, SV/VD luminal pressure directly rose and fell, followed by rhythmic BSM contractions lasting approximately 25s. Acute T8-T9 spinalization (n=4) did not alter the activation pattern of the BSM-EMG response. Injection of the GABA(A)-receptor agonist muscimol, inhibiting neuronal activity into the LSt neuron area after LSt neuron microstimulation (n=5), stopped BSM contractions in midstream. CONCLUSIONS: Electrical microstimulation of LSt neurons activates the entire sequence of ejaculation in rats in a coordinated fashion, ie the emission (SV/VD contraction) followed by expulsion (rhythmic BSM contractions) of living spermatozoa. Midcourse interruption of ejaculation following intraspinal muscimol injection establishes that LSt neurons are the SGE. This could help to identify spinal pharmacological targets for the treatment of ejaculatory disorders and provide the rationale for intraspinal stimulation to treat anejaculation in infertile spinal cord injured (SCI) patients. PMID- 18394783 TI - Training in percutaneous nephrolithotomy--a critical review. AB - AIM: To study factors influencing training and maintaining skills in performing percutaneous nephrolithotomy (PCNL). METHODS: We matched key words, throughout Medline, MeSH, and Cochrane databases including: renal stone, percutaneous, nephrostomy, endourology, educational, training, learning curve, expertise, skill, residency, practice, survey, simulator, and robotics. For this topic we defined, if possible, levels of evidence based on International Consultation on Urological Diseases (ICUD) and World Health Organization recommendations. RESULTS: Obtaining renal access is one of the most important factors in training for PCNL. A resident has to perform about 24 PCNL procedures to obtain a good proficiency during the residence period. Competence at performing PCNL is reached after 60 cases and excellence is obtained at >100 cases. Stone centers providing all the endoscopic treatment options seem to provide the best conditions to ensure a sufficient volume of patients recruited. Virtual reality simulators may have a potential in training for PCNL. To maintain one's expertise, participation in continuing educational programs is recommended. CONCLUSION: PCNL is currently the most complicated stone surgery technique to teach. The steep learning curve is mainly related to obtaining renal access. The traditional method of acquiring surgical skills is by apprenticeship in the absence of validated virtual simulators. Given the complexity of the treatment of renal stones, one may consider a centralized renal stone treatment in dedicated stone centers. PMID- 18394784 TI - The artificial urinary sphincter in patients with spinal cord lesion: description of a modified technique and clinical results. AB - BACKGROUND: The artificial sphincter is the method of choice in patients with stress urinary incontinence due to neurogenic bladder dysfunction. However, long term studies reveal a high revision rate. OBJECTIVE: To determine the success and revision rates of a modified implant. DESIGN, SETTING, AND PARTICIPANTS: In a retrospective analysis, the results of 51 consecutive patients presenting at a private paraplegic center with neurogenic bladder dysfunction (meningomyelocele: n=8; spinal cord injury: n=37; others: n=6) who underwent implantation of an artificial sphincter at the bladder neck using a port instead of a pump were evaluated. MEASUREMENTS: Subjective and objective cure rates were assessed by video-urodynamics and a standardized interview. RESULTS AND LIMITATIONS: After a mean follow up of 95.9 mo, 70.6% of the patients were objectively and subjectively cured; 90.2% were completely continent in everyday life. Mean bladder capacity (465 ml) and compliance (41.7 ml/cm H(2)O) were normal. Sixteen patients underwent 18 revisions (35.3%). One implant had to be permanently removed. This is a single-centre study; thus, the results have to be confirmed. CONCLUSIONS: With a long follow up of 8 yr, the modification presented by our group proved to be highly successful, reliable, safe, and even cost-effective. Therefore, it seems to be a valuable tool for the treatment of this group of patients. PMID- 18394785 TI - It's not a radical prostatectomy, it's a total prostatectomy. PMID- 18394786 TI - Editorial comment on: The artificial urinary sphincter in patients with spinal cord lesion: description of a modified technique and clinical results. PMID- 18394787 TI - Editorial comment on: training in percutaneous nephrolithotomy--a critical review. PMID- 18394788 TI - Editorial comment on: The artificial urinary sphincter in patients with spinal cord lesion: description of a modified technique and clinical results. PMID- 18394789 TI - Endocannabinoid functions controlling neuronal specification during brain development. AB - Endocannabinoids (eCBs) regulate a broad range of physiological functions in the postnatal brain and are implicated in the neuropathogenesis of psychiatric and metabolic diseases. Accumulating evidence indicates that eCB signaling also serves key functions during neurodevelopment; and is inherently involved in the control of neurogenesis, neural progenitor proliferation, lineage segregation, and the migration and phenotypic specification of immature neurons. Recent advances in developmental biology define fundamental eCB-driven cellular mechanisms that also contribute to our understanding of the molecular substrates of prenatal drug, in particular cannabis, actions. Here, we summarize known organizing principles of eCB-signaling systems in the developing telencephalon, and outline the sequence of decision points and underlying signaling pathways upon CB1 cannabinoid receptor activation that contribute to neuronal diversification in the developing brain. Finally, we discuss how these novel principles affect the formation of complex neuronal networks. PMID- 18394790 TI - Difference of the hippocampal and white matter microalterations in MCI patients according to the severity of subcortical vascular changes: neuropsychological correlates of diffusion tensor imaging. AB - OBJECTIVES: Most imaging studies of mild cognitive impairment (MCI) have focused on gray matter alterations, although many MCI patients demonstrate subcortical vascular changes. We investigated the changes of the hippocampal area and various white matter areas in MCI patients with using diffusion tensor imaging (DTI), according to the severity of subcortical vascular changes, and we then correlated the DTI findings with the neuropsychological results. PATIENTS AND METHODS: Among the 40 MCI patients, the 21 non-vascular MCI (nvMCI) and 19 vascular MCI (vMCI) patients were subdivided according to Erkinjuntti's imaging criteria. The mean diffusivity (MD) and fractional anisotropy (FA) were compared in the bilateral temporal, frontal, parietal and occipital white matter regions, as well as in the bilateral hippocampi, centrum semiovale, and the midline genu and splenum of the corpus callosum among the nvMCI and vMCI patients and the 17 controls. The neuropsychological findings were also compared between the subgroups. RESULTS: All the MCI patients showed decreased FA and increased MD in all the regions except the occipital areas. In the parietal regions and centrum semiovale, the vMCI patients had a greater FA decrease than the nvMCI patients and controls. In the hippocampi, the FA was lowest in the nvMCI patients. The memory function in the nvMCI patients was more impaired than that in the vMCI patients. The vMCI patients showed impairment of the visuospatial and frontal executive functions. CONCLUSION: We were able to correlate the microstructural alterations with the neuropsychological findings in the MCI subgroups. PMID- 18394791 TI - Single and combined effects of nickel (Ni(II)) and cobalt (Co(II)) ions on activated sludge and on other aerobic microorganisms: a review. AB - Nickel (N(II)) and cobalt (Co(II)) are often encountered in wastewaters. As conventional wastewater treatment may only partially remove nickel and cobalt, a large fraction of the above metals is released to the aquatic environment. Both metals have been identified as micronutrients, at trace concentrations; however, they are both microbial growth inhibitors, at relatively high concentrations. On the other hand, the combined effects (e.g.: growth stimulation or toxicity) of the above metals have been found to differ from the summation of the effects which occur when the metals are applied individually. Moreover, a number of environmental factors (e.g.: pH, biomedium composition, biomass concentration, presence of other heavy metals) can affect the microbial toxicity of the above metallic species. The present review discusses, in a systematic way, the individual and joint effects of the above heavy metals to the growth of microorganisms grown under aerobic conditions, with focus on the growth of activated sludge. Data on multi-metal toxicity are particularly useful in establishing criteria for heavy metal tolerance levels in the environment. PMID- 18394792 TI - Studies of chromium removal from tannery wastewaters by algae biosorbents, Spirogyra condensata and Rhizoclonium hieroglyphicum. AB - Chromium in the effluent is a major concern for tanning industry. Chemical precipitation methods are commonly employed for the removal of chromium but this leads to formation of chrome-bearing solid waste, plus it is uneconomical when the concentration of chromium in the effluent is low. Ion exchange and membrane separation methods are relatively expensive. In this study, two algae namely, Spirogyra condensata and Rhizoclonium hieroglyphicum have been employed to remove chromium from tannery effluent. The effect of pH and chromium concentration showed S. condensata to exhibit maximum uptake of about 14 mg Cr(III)/g of algae at optimum pH of 5.0 whereas R. hieroglyphicum had 11.81 mg of Cr(III)/g of algae at pH of 4.0. Langmuir and Freundlich models were applied. Increase of initial concentration of Cr resulted to a decrease in adsorption efficiency. Dilute sulphuric acid (0.1M) showed good desorption efficiency (>75%). Interference from cations negatively impacted on biosorption of chromium. Immobilized algae on Amberlite XAD-8 in a glass column, gave better recovery of chromium in tannery effluent compared to a batch method with unimmobilized algae. Fourier transform infra red (FT-IR) analysis of the two algae revealed the presence of carboxyl groups as possible binding sites. PMID- 18394793 TI - Synthesis, structural investigation, thermal decomposition mechanism and sensitivity properties of an energetic compound [Cd(DAT)(6)](ClO(4))(2) (DAT=1,5 diaminotetrazole). AB - An energetic coordination compound [Cd(DAT)(6)](ClO(4))(2) has been synthesized by using 1,5-diaminotetrazole (DAT) as ligand and its structure has been characterized by applying X-ray single crystal diffraction, elemental analysis and FT-IR spectroscopy. The central cadmium(II) cation is coordinated by six N atoms from six DAT molecules to form a six-coordinated and distorted octahedral structure. Di-dimension layer structure was formed and the layers were linked together by the extensive intermolecular hydrogen bonds between DAT ligands and ClO(4)(-) anions. Thermal decomposition mechanism of the title compound was predicted based on DSC, TG-DTG and FT-IR analyses results. The kinetic parameters of the first exothermic process of the title compound were studied by applying the Kissinger's and Ozawa-Doyle's methods. Sensitivity tests reveal that the title compound has sensitive nature. PMID- 18394794 TI - Microbial community dynamics during composting of sewage sludge and straw studied through phospholipid and neutral lipid analysis. AB - The composting process involves a succession of different communities of microorganisms that decompose the initial material, transforming it into a stable final product. In this work, the levels of phospholipid fatty acid (PLFA), neutral lipid fatty acid (NLFA) and sterol were monitored in compost versus time, as indicators of the activity of various microorganisms (Gram-positive or Gram negative bacteria, fungi, etc.). During composting, the PLFA and NLFA from Gram negative bacteria and eukaryotes (2-OH 10; 3-OH 12; 2-OH 14; 13:0; 16:1; 18:1 trans) as well as some sterols of plant origin (e.g. monostearin sterols) decreased until the end of composting. In contrast, the branched fatty acids with iso- and anteiso-forms (i-15:0; a-15:0; i-16; i-17) increased mainly in the thermophilic phase, but decreased right after. The PLFA 18:2 (6; 9), which is used as an index of the occurrence of some fungi, rose strongly at the beginning of composting, but fell after peak heating. In contrast, the other main sterol indicative of fungi, ergosterol, decreased at the beginning of the thermophilic phase, but increased strongly by the end of composting. Accordingly, cluster and PCA analysis separated the PLFA of Gram-negative bacteria and eukaryotic cells from those of Gram-positive bacteria and long-chain fatty acids. The fungal PLFA considered, 18:2 (9, 12), was clustered more closely to iso- and anteiso-branched PLFAs. Stigmasterol, squalene and cholesterol occurred in the lower right part of the loading plot and were clustered more closely to iso-, anteiso-branched PLFAs and 18:2 w 6,9 suggesting their relationship to microbial activities. We also observed the tendency of resistance of fatty acid PLFAs and NLFAs of long chain (19:0 (cis-9); 20:0) and some recalcitrant sterols, e.g. sitosterol, at the end of composting. The presence of high levels of the latter in the final stage indicates their contribution to the structural stability of organic matter fractions. These recalcitrant components were more clustered and occurred in the lower right part of the loading plot. PMID- 18394795 TI - Low-temperature preparation and characterization of iron-ion doped titania thin films. AB - Iron-ion doped titania thin films with an anatase phase were successfully synthesized in this study using the high-pressure crystallization (HPC) process. The crystallization temperature of Fe(3+)-doped TiO(2) thin films was markedly reduced to be as low as 125 degrees C. The films prepared via the HPC process have a more uniform microstructure and smaller grain sizes than the films prepared via the atmospheric-pressure annealing process. The films prepared via both processes were found to have photocatalytic properties under visible light. The films prepared via the HPC process exhibited enhanced photocatalytic activities in comparison with the films annealed via the conventional process. Increasing the annealing temperature in the HPC process resulted in an improvement in the photocatalytic properties because of an increase in the crystallinity of the prepared films. The HPC process was demonstrated to be a potential method for synthesizing visible-light driven titania thin films with enhanced photocatalytic activities at low temperatures. PMID- 18394796 TI - Catalytic properties of carbon materials for wet oxidation of aniline. AB - A mesoporous carbon xerogel with a significant amount of oxygen functional groups and a commercial activated carbon, were tested in the catalytic wet air oxidation of aniline at 200 degrees C and 6.9 bar of oxygen partial pressure. Both carbon materials showed high activity in aniline and total organic carbon removal, a clear increase in the removal efficiency relatively to non-catalytic wet air oxidation being observed. The best results in terms of aniline removal were obtained with carbon xerogel, an almost complete aniline conversion after 1h oxidation with high selectivity to non-organic compounds being achieved. The materials were characterized by thermogravimetric analysis, temperature programmed desorption, N(2) adsorption and scanning electron microscopy, in order to relate their performances to the chemical and textural characteristics. It was concluded that the removal efficiency, attributed to both adsorption and catalytic activity, is related to the mesoporous character of the materials and to the presence of specific oxygen containing functional groups at their surface. The effect of catalytic activity was found to be more important in the removal of aniline than the effect of adsorption at the materials surface. The results obtained indicate that mesoporous carbon xerogels are promising catalysts for CWAO processes. PMID- 18394797 TI - Effect of sulphur on soil Cu/Zn availability and microbial community composition. AB - Successful phytoremediation depends mainly on the bioavailability of copper (Cu) and zinc (Zn) in the soil. We studied the potential effects of sulphur (S) amendment on mobility of copper, zinc and microbial community composition in soil under laboratory conditions. The results showed that with S application at 20 g S kg(-1), soil pH decreased about 3 units and the solubility of the Cu and Zn significantly increased after 64 days of incubation. The concentration of Cu in Cu-accumulator Elsholtzia splendens shoots and roots increased with S treatment. Concentration of Cu in the shoots was 156.5 mg kg(-1) under S treatment. It was 2.5 times of without application of S. PCR-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprint analysis revealed that there were certain groups of acidophilic soil bacteria in the soil after addition of S. We found specific clones such as 1 (from biofilter-treating hydrogen sulfide and methanol) and 4 (from metal-rich and acidic River Tinto) in the soil with S treatment. The above results indicated that S facilitated the mobility of Cu and Zn by soil microorganism and provided a basis for further studies of S-assisted phytoremediation. PMID- 18394798 TI - Kinetic study approach of remazol black-B use for the development of two-stage anoxic-oxic reactor for decolorization/biodegradation of azo dyes by activated bacterial consortium. AB - The laboratory-isolated strains Pseudomonas aeruginosa, Rhodobacter sphaeroides, Proteus mirabilis, Bacillus circulance, NAD 1 and NAD 6 were observed to be predominant in the bacterial consortium responsible for effective decolorization of the azo dyes. The kinetic characteristics of azo dye decolorization by bacterial consortium were determined quantitatively using reactive vinyl sulfonated diazo dye, remazol black-B (RB-B) as a model substrate. Effects of substrate (RB-B) concentration as well as different substrates (azo dyes), environmental parameters (temperature and pH), glucose and other electron donor/co-substrate on the rate of decolorization were investigated to reveal the key factor that determines the performance of dye decolorization. The activation energy (E(a)) and frequency factor (K(0)) based on the Arrhenius equation was calculated as 11.67 kcal mol(-1) and 1.57 x 10(7)mg lg MLSS(-1)h(-1), respectively. The Double-reciprocal or Lineweaver-Burk plot was used to evaluate V(max), 15.97 h(-1) and K(m), 85.66 mg l(-1). The two-stage anoxic-oxic reactor system has proved to be successful in achieving significant decolorization and degradation of azo dyes by specific developed bacterial consortium with a removal of 84% color and 80% COD for real textile effluents vis-a-vis >or=90% color and COD removal for synthetic dye solution. PMID- 18394799 TI - Bioassessment of a combined chemical-biological treatment for synthetic acid mine drainage. AB - In this work, ecotoxicological characteristics of synthetic samples of acid mine drainage (AMD) before and after a combined chemical-biological treatment were investigated by using Lepidium sativum and Daphnia magna. AMD treatment was performed in a two-column apparatus consisting of chemical precipitation by limestone and biological refinement by sulphate reducing bacteria. Synthetic samples of AMD before treatment were toxic for both L. sativum (germination index, G, lower than 10%) and D. magna (100% immobility) due to acid pH and presence of copper and zinc. Chemical treatment (raising pH to 5-6 and eliminating copper) generated effluents with reduced toxicity for L. sativum (G=33%), while 100% immobility was still observed for D. magna. Dynamic trends of toxicity for the first and fifth outputs of the biological column denoted a gradual improvement leading to hormesis for Lepidium (after the initial release of organic excess), while a constant residual toxicity remained for Daphnia (probably due to H(2)S produced by sulphate reducing bacteria). PMID- 18394800 TI - Increased plasma levels of heat shock protein 70 in patients with vascular mild cognitive impairment. AB - Heat shock proteins (Hsps) have been regarded as cytoprotectants that protect brain cells from damage encountered following cerebral ischemia or during the progression of neurodegenerative diseases. In this study, we assessed the plasma Hsp70 and Hsp27 levels in not cognitively impaired (NCI) subjects and in patients with mild cognitive impairment (MCI), vascular mild cognitive impairment (VMCI), and probable Alzheimer's disease (AD). Comparison of the plasma Hsp70 and Hsp27 levels of the 4 groups revealed that only the plasma Hsp70 level of VMCI patients (14.11 ng/ml) was significantly higher than that of NCI subjects (11.32 ng/ml), MCI patients (10.16 ng/ml), and patients with probable AD (10.16 ng/ml) after adjustment of age, sex, and education (F=4.231, d.f.=3, p=0.008). Furthermore, there was no difference in plasma Hsp27 levels among the 4 groups. These findings suggest that the plasma Hsp70 level may be related to vascular factors or inflammation. PMID- 18394801 TI - Enhanced Th2 immunity after DNA prime-protein boost immunization with amyloid beta (1-42) plus CpG oligodeoxynucleotides in aged rats. AB - Generation and accumulation of fibrillar amyloid beta (Abeta) is widely considered as the pathogenic basis of neurodegeneration in Alzheimer's disease (AD). Both active immunization with fibrillar Abeta and passive immunization with anti-Abeta antibodies in transgenic mouse models of AD result in prevention/dissociation of Abeta plaque formation and restoration of cognitive functions. However, similar immunization studies in humans had to be halted because 6% of the AD patients developed acute meningoencephalitis, likely due to anti-Abeta specific autoimmune Th1 cells. Hence, making Abeta immunotherapy successful requires production of strong antibody responses without Th1-type immunity. In an attempt to develop safer vaccines, we examined the influence of oligodeoxynucleotides as adjuvant on the Th1 and Th2 immune response to Abeta in aged rats. We further investigated whether a DNA prime-protein boost strategy could elicit a more robust Th2 response. The results of the present study showed that all the animals injected with either Abeta peptide alone or Abeta encoding plasmid alone or plasmid DNA prime followed by peptide boost have elicited specific anti-Abeta antibodies. When co-administered, synthetic oligodeoxynucleotides (ODN) further enhanced the anti-Abeta titres. More importantly, the IgG subclasses of the antibodies generated by DNA prime-peptide boost regimen with ODN as adjuvant were primarily of IgG2b and IgG1 isotypes, suggesting that heterologous immunization strategy along with ODN would be advantageous in eliciting more beneficial Th2-type humoral immune response. PMID- 18394802 TI - Vesicular acetylcholine transporter knock-down mice are more susceptible to pilocarpine induced status epilepticus. AB - The pilocarpine (PILO) animal model of Temporal Lobe Epilepsy (TLE) portrays the most common changes in hippocampal circuitry found in human TLE. The acute cholinergic insult induces status epilepticus (SE), which triggers an overwhelming set of plastic events that result on late spontaneous recurrent limbic seizures. It has been suggested that the cholinergic system plays an important role in the synchronization required for ictogenesis. We took advantage of a knock-down animal model for the vesicular acetylcholine transporter (VAChT KD) to investigate seizure genesis in a model of cholinergic dysfunction. We induced SE in VAChT KD and wild-type (WT) mice by a single intraperitoneal injection of PILO in order to evaluate susceptibility to seizures. Video-EEG recordings evaluated epileptiform activity and ictal behavior onset. The hypothesis tested is that innate cholinergic hypofunction could result in increased susceptibility to PILO. VAChT KD(HOM) mice showed shorter latency for the first epileptiform discharge and for the first seizure episode, when compared to other groups. The duration of these seizure episodes, however, were not statistically different among experimental groups. On the other hand, VAChT KD(HOM) had the shortest latency to isoelectric EEG, when compared to WT and KD(HET). Our results indicate that a reduction of brain VAChT protein to the levels found in VAChT KD(HOM) mice alters the epileptic response to PILO. Thus, fine-tuning modulation of cholinergic tone can affect the susceptibility of epileptic responses to pilocarpine. PMID- 18394803 TI - Transient early expression of TNF-alpha in sciatic nerve and dorsal root ganglia in a mouse model of painful peripheral neuropathy. AB - The proinflammatory cytokine tumor necrosis factor-alpha (TNF) is an important mediator in neuropathic pain. We investigated the temporal pattern of TNF mRNA expression in the sciatic nerve, in dorsal root ganglia (DRG) and spinal cord in the mouse chronic constriction injury model of neuropathy with quantitative real time polymerase chain reaction. Neuropathic pain-like behaviour was monitored by evaluating thermal hyperalgesia and mechanical allodynia. Pain-related behaviour and TNF expression were evaluated 6 h, 1, 3, 7 and 14 days after injury. Naive animals and sham-operated mice were used as controls. We found an early upregulation of sciatic nerve TNF mRNA levels in chronic constriction injury (CCI) and sham-operated animals 6 h after surgery: 1 day later TNF overexpression was present in CCI mice only and disappeared 3 days after injury. The mRNA cytokine levels were elevated in DRG 1 and 3 days after surgery in CCI animals only, while the cytokine was not modulated in the spinal cord. A significant hyperalgesia was present in CCI and sham-operated mice at 6 h and 1 day, while at later time point only CCI mice presented lower thresholds. Mechanical allodynia was already present only in CCI animals 6 h from surgery and remained constant up to the 14 th day. The results indicate that a transient early TNF upregulation takes place in peripheral nervous system after CCI that can activate a cascade of proinflammatory/pronociceptive mediators. PMID- 18394804 TI - Changes in mu rhythm during action observation and execution in adults with Down syndrome: implications for action representation. AB - The human mirror neuron system is thought to be the underlying basis of perception-action coupling involved in imitation and action understanding. In order to examine this issue we examined the recruitment of the mirror neuron system, as reflected in mu rhythm suppression in a population of adults with Down syndrome (DS) with known strengths in imitation but with impairments in perceptual-motor coupling. Ten healthy adults and 10 age-matched adults with (DS) participated in the study. Subjects were asked to make self-paced movements (execution), and view movements made by the experimenter (observation). The action consisted of reaching with the dominant hand to grasp and lift a cup. Cortical responses were recorded with a whole head magnetoencephalography (MEG) system. Both groups demonstrated significant attenuation of the mu rhythm in bilateral sensorimotor areas when executing the action. Typical adults also demonstrated significant mu suppression in bilateral sensorimotor areas during observation of the action. In contrast, when observing the movement, adults with DS showed a significantly reduced overall attenuation of mu activity with a distinct laterality in the pattern of mu suppression. These results suggest that there is a dysfunction in the execution/observation matching system in adults with DS and has implications for the functional role of the human mirror neuron system. PMID- 18394805 TI - Sex-specific pain modulation: the growth factor, neuregulin-1, as a pro nociceptive cytokine. AB - An increasing amount of evidence indicates that there are significant sex differences in clinical and experimental pain sensitivity in men and women. While it is now clear that the endogenous sex steroids are involved in mediating these sex differences, the cellular and molecular mechanisms that underlie their effects on nociceptive sensitivity remain elusive. Recent studies have shown that sex steroids are potent regulators of gene expression in glial cells, particularly astrocytes. This review specifically highlights some of the evidence of sex steroid regulation of growth factor expression. Growth factors have been shown to be potent pro-nociceptive mediators in rodents. Thus, regulation of their expression by sex steroids may be a general mechanism by which sex steroids exert their effect on pain sensitivity. One such mechanism, the progesterone specific regulation of the growth factor, neuregulin-1, following nerve root injury in the rat, is described in detail. Neuregulin-1 expression is increased in spinal cord astrocytes only in female rats with circulating progesterone. Neuregulin-1 has also been shown to produce transient tactile allodynia when delivered intrathecally in rats. Our understanding of growth factor regulation by sex steroids promises to open up new avenues of investigation into the mechanisms that drive sex differences in pain sensitivity. PMID- 18394806 TI - Estimating the clinical effectiveness of cognitive behavioural therapy in the clinic: evaluation of a CBT informed pain management programme. AB - Randomized controlled trials and meta-analyses provide evidence for the efficacy of cognitive-behaviourally informed treatment (CBT) programmes for chronic pain. The current study aims to provide practice-based evidence for the effectiveness of CBT in routine clinical settings. Over a 10-year period 1013 pain patients were accepted into a 4 week in-patient pain management programme. Data from more than 800 patients was available at pre-treatment and at one month post-treatment and for around 600 patients at pre-treatment and at 9 months follow-up. Measures reported in this analysis were pain experience and interference, psychological distress (depression and anxiety), self-efficacy, catastrophizing, and walking. Change from pre-treatment to post-treatment and follow-up was assessed with conventional statistical tests, the computation of effect sizes and by the reliable change index (RCI) and clinically significant change (CSC) methodology. These analyses provide evidence of statistical improvement at post-treatment and follow-up and the RCI/CSC methodology suggested that between 1 in 3 and 1 in 7 (depending on the outcome measure) achieved clinically significant gains. There was also evidence that a small percentage of patients (1-2%) reliably deteriorated during the period of treatment. The limitations in the inferences that can be drawn from this study and of the methodology are discussed. A case is made for the application of benchmarking methods using data from RCTs in order to more fully evaluate practice and to generate better quality practice based evidence. PMID- 18394807 TI - Comparative experimental Haemonchus contortus infection of two sheep breeds native to the Canary Islands. AB - This study compares the susceptibility to Haemonchus contortus infection in two breeds of sheep endemic to the Canary Islands, the Canaria Hair Breed sheep and the Canaria sheep. Sheep were experimentally infected with 20,000 larvae of H. contortus and animals killed on days 7 and 28 post-infection. No difference between sheep breeds were detected in immature worm counts at days 7 or 28 post infection. However, in comparison to the Canaria sheep breed, the Canaria Hair Breed sheep showed lower mean faecal egg counts, lower adult worm counts, lower number of eggs in utero and female worm stunting. Overall, these data suggest that the Canaria Hair Breed sheep has a greater resistance to H. contortus infection than Canaria sheep, and that this resistance may act at the level of the adult parasite. PMID- 18394808 TI - Distribution of Cryptosporidium parvum subtypes in calves in Germany. AB - Cryptosporidium DNA was extracted from 134 faecal specimens from pre-weaned calves from different German Federal States (age range, 3-15 days old), which tested positive for oocysts by microscopic analysis. The 18S rDNA gene and the oocyst wall protein gene (COWP) were used as targets for PCR and RFLP techniques. Cryptosporidium species were identified by using SspI, MboII and RsaI endonucleases for the digestion of 18S rDNA and COWP amplified fragments, respectively. In all samples, restriction patterns corresponding to Cryptosporidium parvum were identified, which is in agreement with abundant literature data indicating C. parvum as the most common species in pre-weaned calves. In order to estimate the genetic heterogeneity among C. parvum calf isolates, 53 samples chosen to represent different German Federal States were successfully subtyped by sequence analysis of the highly polymorphic 60-kDa glycoprotein gene. All isolates belonged to the allele IIa (with seven subtypes), with the exception of one isolate that belonged to the allele IId. Moreover, three novel subtypes of the allele family IIa have been found. This study confirms the utility of genotyping and subtyping tools in characterizing the transmission of Cryptosporidium spp. This is the first molecular epidemiological report about subtyping of Cryptosporidium bovine isolates in Germany. PMID- 18394809 TI - Experimental infection of cats (Felis catus) with Tritrichomonas foetus isolated from cattle. AB - Tritrichomonas foetus is recognized as the causative agent of venereal trichomoniasis in cattle. It is characterized by embryonic and early fetal death and post-coital pyometra, and feline trichomoniasis, manifest as chronic, large bowel diarrhea. Many of the infected cats are less than 2 years old and specific routes of transmission remain unknown. We recently demonstrated that feline isolates of T. foetus can successfully infect heifers, resulting in pathologic changes similar, but not identical to those previously reported as representative of bovine trichomoniasis. In this study, we experimentally infected six cats less than 1 year of age with a bovine (D-1) isolate of T. foetus and one cat with a feline (AUTf-1) isolate of T. foetus. Within 2 weeks, the cat infected with the feline (AUTf-1) isolate was culture positive for trichomonads in weekly fecal samples. At the end of 5 weeks, only one cat infected with the bovine (D-1) isolate was fecal culture positive for trichomonads. At necropsy, the intestine of each cat was removed and divided into five sections (ileum, cecum, anterior, medial and posterior colon). Contents from each section were collected and cultured. The cat infected with the feline (AUTf-1) isolate was culture positive in the ileum, cecum, medial and posterior colon. Two cats infected with the bovine (D-1) isolate were culture positive in the cecum only. Additionally, each intestinal section was submitted to a pathologist for histopathological examination. The combined results indicate that there are demonstrable differences between the feline (AUTf-1) and bovine (D-1) isolates regarding their infectivity in cats. PMID- 18394810 TI - Epizootics of metazoan gill parasites did not threaten feasibility of farming southern bluefin tuna (Thunnus maccoyii) in a trial extending over summer months. AB - Tuna farming off Port Lincoln, Australia, involves catching wild 2-4-year-old southern bluefin tuna in summer and then fattening for periods of 2-8 months. As fresh product is not available year-round, the feasibility of maintaining tuna for longer periods was trialled, including over a summer season, when temperatures may exceed 24 degrees C. As the rates of growth and reproduction in ectoparasites of fishes are usually most rapid during warm temperatures, parasite epizootics at this time may adversely affect the health of tuna. We collected epidemiological data on burdens of metazoans on the gills of tuna from the time of stocking in April 2005 through to final harvest in August 2006 (N=220). We document an epizootic of the copepod Pseudocycnus appendiculatus, characterised by a significant increase in both prevalence and mean intensity in the first winter, followed by a decline in these parameters over the next 12 months. This epizootic pattern appears to be independent of seasonal changes in temperature. For two other species, a second copepod (Euryphorus brachypterus) and a polyopisthocotylean flatworm (Hexostoma thynni), there were no clearly discernible trends in infections. As the high water temperatures over the summer period did not lead to increased infections of any species of gill parasites, we conclude that they do not threaten the feasibility of farming of Thunnus maccoyii. PMID- 18394811 TI - Zinc supplementation increases resistance to experimental infection by Trypanosoma cruzi. AB - It is well recognized that zinc is an essential trace element for all organisms, influencing growth and affecting the development and integrity of the immune system. It is also well known that the protective response against Trypanosoma cruzi depends on both innate and acquired immunity and for the control of the parasite load and host survival, the participation of special cells such natural killer (NK), T and B lymphocytes and macrophages are required. So the aims of this study were to evaluate the effects of zinc supplementation on the host's immune response infected with T. cruzi. Our data point in the direction that zinc supplementation triggered enhanced thymocyte and splenocyte proliferation as compared to unsupplied group of animals. It is also important to emphasize that interleukin-12 (IL-12) participates in the resistance to several intracellular pathogens including T. cruzi. Our findings demonstrate an enhanced production of IL-12 during the acute phase of infection in zinc-supplied groups. So we conclude that zinc supplementation leads to an effective host's immune response by up modulating the host's immune response, thus contributing in the reduction of blood parasites and the harmful pathogenic effects of the experimental Chagas' disease. PMID- 18394812 TI - Chronic psychosocial stress affects corticotropin-releasing factor in the paraventricular nucleus and central extended amygdala as well as urocortin 1 in the non-preganglionic Edinger-Westphal nucleus of the tree shrew. AB - Stressful stimuli evoke neuronal and neuroendocrine responses helping an organism to adapt to changed environmental conditions. Chronic stressors may induce maladaptive responses leading to psychiatric diseases, such as anxiety and major depression. A suitable animal model to unravel mechanisms involved in the control of adaptation to chronic stress is the psychological subordination stress in the male tree shrew. Subordinate male tree shrews exhibit chronic hypothalamo pituitary-adrenal (HPA) activation as reflected in continuously elevated cortisol secretion, and structural changes in the hippocampal formation. Corticotropin releasing factor (CRF) is the major peptide released upon activation of the HPA axis in response to stress. Recent evidence suggests that besides CRF, urocortin 1 (Ucn1) also plays a role in stress adaptation. We have tested the significance of CRF and Ucn1 in adaptation to chronic psychosocial stress in male tree shrews exposed for 35 days to daily psychosocial conflict, by performing semi quantitative immunocytochemistry for CRF in the parvocellular hypothalamic paraventricular nucleus (pPVN), extended amygdala, viz. central extended amygdala (CeA) and dorsolateral nucleus of the bed nucleus of the stria terminalis (BNSTdl) as well as that for Ucn1 in the non-preganglionic Edinger-Westphal nucleus (npEW). Compared to unstressed animals, psychosocial stress resulted in an immediate and sustained activation of the HPA axis and sympathetic tone as well as reduced testosterone concentration and decreased body and testis weights vs. non-stressed tree shrews. In the pPVN, the number of CRF-immunoreactive neurons and the specific signal density of CRF-immunoreactive fiber terminals in the CeA were strongly reduced (-300 and -40%, respectively; P<0.05), whereas no significant difference in CRF fiber density was found in BNSTdl. The npEW revealed 4 times less Ucn1-immunoreactive neurons (P<0.05). These clear effects on both Ucn1- and CRF-neuropeptide contents may reflect a crucial mechanism enabling the animal to adapt successfully to the stressors, and point to the significance of the pPVN, CeA and npEW in stress-induced brain diseases. PMID- 18394813 TI - Does lower response to alcohol explain the high co-morbidity between bipolar disorder and alcohol dependence? AB - The frequency of the co-occurrence between alcohol use disorders and bipolar disorder is well documented. However, the etiology of this association is discussed. Among the most quoted hypothesis, high alcohol consumption has been considered as a form of self-medication. At variance with this assumption, we hypothesized that mania is associated with a low level of response to alcohol. Indeed, a low level of response to alcohol was previously associated with an increased risk of alcohol dependence in a high risk population. We discuss the neurobiological findings supporting our assumption, including the decreased reward-related brain activation in mania and further propose a brief study design to test this hypothesis. PMID- 18394814 TI - Comparison of FDG-PET/CT and CT for delineation of lumpectomy cavity for partial breast irradiation. AB - PURPOSE: The success of partial breast irradiation critically depends on proper target localization. We examined the use of fluorodeoxyglucose-positron emission tomography (FDG-PET)/computed tomography (CT) for improved lumpectomy cavity (LC) delineation and treatment planning. METHODS AND MATERIALS: Twelve breast cancer patients underwent FDG-PET/CT on a GE Discovery scanner with a median time from surgery to PET/CT of 49 days. The LC was contoured on the CT scan by a radiation oncologist and, together with a nuclear medicine physician, on the PET/CT scan. The volumes were calculated and compared in each patient. Treatment planning target volumes (PTVs) were calculated by expanding the margin 2 cm beyond the LC, maintaining a 5-mm margin from the skin and chest wall, and the treatment plans were evaluated. In addition, a study with a patient-like phantom was conducted to evaluate the effect that the window/level settings might have on contouring. RESULTS: The margin of the LC was well visualized on all FDG-PET images. The phantom results indicated that the difference between the known volume and the FDG-PET-delineated volume was <10%, regardless of the window/level settings. The PET/CT volumes were larger than the CT volumes in all cases (median volume ratio, 1.68; range, 1.24-2.45; p = 0.004). The PET/CT-based PTVs were also larger than the CT-based PTV (median volume ratio, 1.16; range, 1.08-1.64; p = 0.006). In 9 of 12 patients, a CT-based treatment plan did not provide adequate coverage of the PET/CT-based PTV (99% of the PTV received <95% of the prescribed dose), resulting in substantial cold spots in some plans. In these cases, treatment plans were generated which were specifically designed to cover the larger PET/CT based PTV. Although these plans showed an increased dose to the normal tissues, the increases were modest: the non-target breast volume receiving > or =50 Gy, lung volume receiving > or =30 Gy, and heart volume receiving > or =5 Gy increased by 5.7%, 0.8%, and 0.2%, respectively. The normal tissue dose-volume objectives were still met with these plans. CONCLUSION: The results of our study have shown that FDG-PET/CT can be used to define the LC volume. The increased FDG uptake was likely a result of postoperative inflammation in the LC. The targets defined using PET/CT were significantly larger than those defined with CT alone. Our results have shown that treatment plans can be generated to cover these larger PET/CT target volumes with only a modest increase in irradiated tissue volume compared with CT-determined PTVs. PMID- 18394815 TI - Tangential radiotherapy without axillary surgery in early-stage breast cancer: results of a prospective trial. AB - PURPOSE: To determine the risk of regional-nodal recurrence in patients with early-stage, invasive breast cancer, with clinically negative axillary nodes, who were treated with breast-conserving surgery, "high tangential" breast radiotherapy, and hormonal therapy, without axillary surgery or the use of a separate nodal radiation field. METHODS AND MATERIALS: Between September 1998 and November 2003, 74 patients who were >/=55 years of age with Stage I-II clinically node-negative, hormone-receptor-positive breast cancer underwent tumor excision to negative margins without axillary surgery as a part of a multi-institutional prospective study. Postoperatively, all underwent high-tangential, whole-breast radiotherapy with a boost to the tumor bed, followed by 5 years of hormonal therapy. RESULTS: For the 74 patients enrolled, the median age was 74.5 years, and the median pathologic tumor size was 1.2 cm. Lymphatic vessel invasion was present in 5 patients (7%). At a median follow-up of 52 months, no regional-nodal failures or ipsilateral breast recurrences had been identified (95% confidence interval, 0-4%). Eight patients died, one of metastatic disease and seven of other causes. CONCLUSION: In this select group of mainly older patients with early-stage hormone-responsive breast cancer and clinically negative axillary nodes, treatment with high-tangential breast radiotherapy and hormonal therapy, without axillary surgery, yielded a low regional recurrence rate. Such patients might be spared more extensive axillary treatment (axillary surgery, including sentinel node biopsy, or a separate nodal radiation field), with its associated time, expense, and morbidity. PMID- 18394816 TI - concomitant chemoradiotherapy with mitomycin C and cisplatin in advanced unresectable carcinoma of the head and neck: phase I-II clinical study. AB - PURPOSE: To evaluate the toxicity and efficacy of concomitant chemoradiotherapy with mitomycin C and cisplatin in the treatment of advanced unresectable squamous cell carcinoma of the head and neck. PATIENTS AND METHODS: Treatment consisted of conventional radiotherapy (70 Gy in 35 fractions), mitomycin C 15 mg/m(2) IV, applied after the delivery of 10 Gy, and cisplatin at an initial dose of 10 mg/m(2)/d IV, applied during the last 10 fractions of irradiation ("chemoboost"). The cisplatin dose was escalated with respect to the toxic side effects by 2 mg/m(2)/d up to the maximum tolerated dose (MTD) or at the most 14 mg/m(2)/d (Phase I study), which was tested in the subsequent Phase II study. RESULTS: All 36 patients had Stage T4 and/or N3 disease, and the majority had oropharyngeal (50%) or hypopharyngeal (39%) primary tumors. Six patients were treated at each of the three cisplatin dose levels tested (Phase I study). Dose-limiting toxicity was not reached even at 14 mg/m(2)/d of cisplatin, which was determined as the MTD and tested in an additional 18 patients (Phase II study). After a median follow-up time of 48 months, 4-year locoregional control, failure-free, and overall survival rates were 30%, 14%, and 20%, respectively. In 24 patients treated at the cisplatin dose level of 14 mg/m(2)/d, the corresponding rates were 40%, 20%, and 22%, respectively. CONCLUSION: Concomitant chemoradiotherapy with mitomycin C and cisplatin "chemoboost" at 14 mg/m(2)/d is feasible, with encouraging survival results if the extremely poor disease profile of the treated patients is considered. PMID- 18394817 TI - Bronchoscopic implantation of a novel wireless electromagnetic transponder in the canine lung: a feasibility study. AB - PURPOSE: The success of targeted radiation therapy for lung cancer treatment is limited by tumor motion during breathing. A real-time, objective, nonionizing, electromagnetic localization system using implanted electromagnetic transponders has been developed (Beacon electromagnetic transponder, Calypso Medical Technologies, Inc., Seattle, WA). We evaluated the feasibility and fixation of electromagnetic transponders bronchoscopically implanted in small airways of canine lungs and compared to results using gold markers. METHODS AND MATERIALS: After approval of the Animal Studies Committee, five mongrel dogs were anesthetized, intubated, and ventilated. Three transponders were inserted into the tip of a plastic catheter, passed through the working channel of a flexible bronchoscope, and implanted into small airways of a single lobe using fluoroscopic guidance. This procedure was repeated for three spherical gold markers in the opposite lung. One, 7, 14, 28, and 60 days postimplantation imaging was used to assess implant fixation. RESULTS: Successful bronchoscopic implantation was possible for 15 of 15 transponders and 12 of 15 gold markers; 3 markers were deposited in the pleural space. Fixation at 1 day was 15 of 15 for transponders and 12 of 12 for gold markers. Fixation at 60 days was 6 of 15 for transponders and 7 of 12 for gold markers, p value = 0.45. CONCLUSIONS: Bronchoscopic implantation of both transponders and gold markers into the canine lung is feasible, but fixation rates are low. If fixation rates can be improved, implantable electromagnetic transponders may allow improved radiation therapy for lung cancer by providing real-time continuous target tracking. Developmental work is under way to improve the fixation rates and to reduce sensitivity to implantation technique. PMID- 18394818 TI - Intraoperative radiation therapy for locally advanced and recurrent soft-tissue sarcomas in adults. AB - PURPOSE: To analyze the outcomes of and identify prognostic factors for patients treated with surgery and intraoperative radiotherapy (IORT) for locally advanced and recurrent soft-tissue sarcoma in adults from a single institution. METHODS AND MATERIALS: We retrospectively reviewed 50 consecutive patients treated with IORT to 62 sites of disease. Primary sites included retroperitoneum-pelvis (78%), extremity (8%), and other (14%). Seventy percent of patients had recurrent disease failing prior surgery (70%) and/or radiation (32%). Mean disease-free interval (DFI) before IORT was 1.9 years (range, 2 weeks-5.4 years). The IORT was delivered with orthovoltage X-rays using individually sized beveled cone applicators. Clinical characteristics were as follows: mean tumor size, 10 cm (range, 1-25 cm); high-grade histologic subtype (72%); and mean dose, 1,159 cGy (range, 600-1,600 cGy). Postoperative radiation or chemotherapy was administered to 37% of IORT Sites and 32% of patients, respectively. Outcomes measured were infield control (IFC), locoregional control (LRC), distant metastasis-free survival (DMFS), disease-specific survival (DSS), and treatment-related complications. Mean and median follow-up of alive patients were 59 and 35 months, respectively. RESULTS: Kaplan-Meier 5-year IFC, LRC, DMFS, and DSS probabilities for the entire group were 55%, 26%, 51%, and 25%, respectively. Prognostic factors found to be significant (p < 0.05) on multivariate analysis were prior DFI and tumor size for LRC, extremity location and leiomyosarcoma histologic subtype for DMFS, and prior DFI for DSS. Our cohort had five Grade 3/4 complications associated with treatment or a 5-year Kaplan-Meier Grade 3/4 complication-free survival rate of 85%. CONCLUSIONS: IORT after tumor reductive surgery is well tolerated and seems to confer IFC in carefully selected patients. PMID- 18394819 TI - Baseline pulmonary function as a predictor for survival and decline in pulmonary function over time in patients undergoing stereotactic body radiotherapy for the treatment of stage I non-small-cell lung cancer. AB - PURPOSE: To examine the effect of baseline forced expiratory volume in 1 second (FEV(1)) and diffusion capacity for carbon monoxide (Dl(co)) on posttreatment survival and pulmonary function decrease after stereotactic body radiotherapy (SBRT) for patients with early-stage non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: Seventy medically inoperable patients with Stage I NSCLC were treated with definitive SBRT to a dose of 6,000 (Stage IA) or 6,600 cGy (Stage IB), given in three equal fractions. Baseline and serial posttreatment pulmonary function data were collected. RESULTS: Median age was 70.5 years, and median follow-up was 2.17 years. Median pretreatment FEV(1) and Dl(co) were 1.05 L and 10.06 mg/min/mm Hg, respectively. There was no significant decrease in survival in patients with baseline FEV(1) and Dl(co) less than the median value and less than the lowest quartile, whereas patients with values greater than the highest quartile of baseline FEV(1) had significantly inferior survival. There was no significant effect of pretreatment FEV(1) or Dl(co) on posttreatment levels. There was a statistically significant decrease in Dl(co) of 1.11 mg/min/mm Hg/y. CONCLUSIONS: Poor baseline pulmonary function did not predict decreased survival or pulmonary function after treatment. A statistically significant decrease in Dl(co) after treatment was seen, similar to decreases seen in studies delivering standard thoracic radiotherapy. We conclude that low pretreatment FEV(1) and/or Dl(co) alone should not be used to exclude patients with NSCLC from treatment with SBRT. PMID- 18394820 TI - [Human monkeypox]. AB - Unlike other recent viral emergences, which were in majority caused by RNA viruses, the monkeypox results from infection by a DNA virus, an orthopoxvirus closely related to both vaccine and smallpox viruses and whose two genomic variants are known. Unexpectedly isolated from captive Asiatic monkeys and first considered as an laboratory curiosity, this virus was recognised in 1970 as an human pathogen in tropical Africa. Here it was responsible for sporadic cases following intrusions (for hunting) into tropical rain forests or rare outbreak with human-to-human transmission as observed in 1996 in Democratic Republic of Congo. As monkeypox in humans is not distinguishable from smallpox (a disease globally eradicated in 1977) it was only subjected to vigilant epidemiological surveillance and not considered as a potential threat outside Africa. This point of view radically changed in 2003 when monkeypox was introduced in the USA by African wild rodents and spread to 11 different states of this country. Responsible for 82 infections in American children and adults, this outbreak led to realize the sanitary hazards resulting from international trade of exotic animals and scientific investigations increasing extensively our knowledge of this zoonosis. PMID- 18394821 TI - [Treatment of venous thromboembolic disease in cancer patients]. AB - Venous thromboembolism (VTE) disease, as defined by the occurrence of deep venous thrombosis or pulmonary embolism, occurs among 4 to 20% of patients with cancer and is a leading cause of death among these patients. Use of classical anticoagulation to treat VTE in a cancer patient is associated with a higher risk of major bleeding and of VTE recurrence as compared to noncancer patients. Updated comprehensive and systematic review of current data from the medical literature allows to reconsider the classical approach used for anticoagulant treatment in cancer patients and to implement adapted recommendations. In 2008, the use of daily subcutaneous low-molecular-weight heparin (LMWH) for at least three to six months is recommended as first line therapy to treat VTE disease in cancer patients. If LMWH are contra-indicated (renal insufficiency), other therapeutic approaches are warranted, such as use of unfractionated heparin (UFH) with early introduction of anti-vitamin K for at least three months or venous cava filter in case of absolute contra-indications to anticoagulation. VTE prophylaxis in cancer patients relies on the same therapeutic approaches as currently used for noncancer patients at high risk of VTE. The definition of more specific prophylactic approaches for patients with cancer considered at higher risks of VTE, will be the subject of many clinical trials in the forthcoming years. PMID- 18394822 TI - [New antitumor agent: In vitro activity on breast carcinoma cells]. AB - Previous work showed that established interactions between water-soluble polymers and cell membrane receptors can lead to modulate cell proliferation and differentiation in vitro. These polymers can be considered as bioactive. The aim of this work was to establish the consequences of the interactions between human breast cancer cells MCF7 and polymers of various chemical compositions regarding cell adhesion and proliferation onto tissue culture plate. Water soluble copolymers were synthesized by radical polymerization and are composed of methacrylic acid and sodium styrene sulphonate units. The modulation of the MCF7, biological-induced by these polymers of various compositions, was evaluated. The influence of the polymers chemical composition on the kinetics of cell proliferation, as well as cell morphology and spreading, were studied. A polymer concentration-dependent inhibition effect was observed. One hundred microgram per liter polymers solutions induced strong inhibition of cell proliferation, as well as a change of the MCF7 cells morphology, which can be related to an inhibition of cell spreading. The polymers/MCF7 cells interactions are modulated by the chemical composition of the copolymers and then the respective rate in sulphonate and carboxylate groups distributed along the macromolecular chain. PMID- 18394824 TI - Ultradian activity rhythms in large groups of newly hatched chicks (Gallus gallus domesticus). AB - A clutch of young chicks housed with a mother hen exhibit ultradian (within day) rhythms of activity corresponding to the brooding cycle of the hen. In the present study clear evidence was found of ultradian activity rhythms in newly hatched domestic chicks housed in groups larger than natural clutch size without a mother hen or any other obvious external time-keeper. No consistent synchrony was found between groups housed in different pens within the same room. The ultradian rhythms disappeared with time and little evidence of group rhythmicity remained by the third night. This disappearance over time suggests that the presence of a mother hen may be pivotal for the long-term maintenance of these rhythms. The ultradian rhythm of the chicks may also play an important role in the initiation of brooding cycles during the behavioural transition of the mother hen from incubation to brooding. Computer simulations of individual activity rhythms were found to reproduce the observations made on a group basis. This was achievable even when individual chick rhythms were modelled as independent of each other, thus no assumptions of social facilitation are necessary to obtain ultradian activity rhythms on a group level. PMID- 18394823 TI - Enterococcus faecium isolated from bone marrow transplant patients in Tunisia: high prevalence of antimicrobial resistance and low pathogenic power. AB - OBJECTIVES: Investigation of the occurrence and antibiotic susceptibility of Enterococcus faecium isolates, collected during four years from neutropenic patients at the Tunisian bone marrow transplantation centre. MATERIALS AND METHODS: E. faecium strains were identified by conventional methods and by the Api20 Strep (Bio-Merieux, France). Antibiotic susceptibility was determined by the disk diffusion method on Mueller-Hinton agar and interpreted as recommended by CA-SFM. MICs of ampicillin, vancomycin, and teicoplanin were determined by E test method. RESULTS: Two hundred and thirty five E. faecium isolates were recovered from stool cultures or rectal swabs (229), throat (three), urine (two), and pus of wound (one). None was responsible for bacteraemia. Ampicillin resistance, without production of beta-lactamase, was observed in 43.8% of isolates. All the isolates were susceptible to glycopeptides. High rates of resistance were observed: high-level resistance (HLR) to gentamicin (33.6%), HLR to kanamycin (55.7%), HLR to streptomycin (47.6%), erythromycin (86.4%), ciprofloxacin (78.7%), rifampicin (85%), and tetracycline (43%). Strains with HLR to gentamicin were significantly more resistant to ampicillin and streptomycin. Multiple drug resistance was observed in most isolates. CONCLUSION: These findings demonstrated the low pathogenic power of E. faecium in our patients, and the high frequencies of resistance to ampicillin and aminoglycosides. In the absence of glycopeptide-resistance, vancomycin remains an alternative treatment against multidrug resistant strains. PMID- 18394825 TI - Human choice among five alternatives when reinforcers decay. AB - Human participants played a computer game in which choices among five alternatives were concurrently reinforced according to dependent random-ratio schedules. "Dependent" indicates that choices to any of the wedges activated the random-number generators governing reinforcers on all five alternatives. Two conditions were compared. In the hold condition, once scheduled, a reinforcer - worth a constant five points - remained available until it was collected. In the decay condition, point values decreased with intervening responses, i.e., rapid collection was differentially reinforced. Slopes of matching functions were higher in the decay than hold condition. However inter-subject variability was high in both conditions. PMID- 18394826 TI - A novel SINE family occurs frequently in both genomic DNA and transcribed sequences in ixodid ticks of the arthropod sub-phylum Chelicerata. AB - Reassociation kinetics and flow cytometry data indicate that ixodid tick genomes are large, relative to most arthropods, containing>or=10(9) base pairs. The molecular basis for this is unknown. We have identified a novel small interspersed element with features of a tRNA-derived SINE, designated Ruka, in genomic sequences of Rhipicephalus appendiculatus and Boophilus (Rhipicephalus) microplus ticks. The SINE was also identified in expressed sequence tag (EST) databases derived from several tissues in four species of ixodid ticks, namely R. appendiculatus, B. (R.) microplus, Amblyomma variegatum and also the more distantly related Ixodes scapularis. Secondary structure predictions indicated that Ruka could adopt a tRNA structure that was, atypically, most similar to a serine tRNA. By extrapolation the frequency of occurrence in the randomly selected BAC clone sequences is consistent with approximately 65,000 copies of Ruka in the R. appendiculatus genome. Real time PCR analyses on genomic DNA indicate copy numbers for specific Ruka subsets between 5800 and 38,000. Several putative conserved Ruka insertion sites were identified in EST sequences of three ixodid tick species based on the flanking sequences associated with the SINEs, indicating that some Ruka transpositions probably occurred prior to speciation within the metastriate division of the Ixodidae. The data strongly suggest that Class I transposable elements form a significant component of tick genomes and may partially account for the large genome sizes observed. PMID- 18394827 TI - Assessment of suitability of two serotype A candidate vaccine strains for inclusion in FMD vaccine in India. AB - The recent type A foot and mouth disease virus field isolates recovered in India are shown to be antigenically quite divergent from the in-use vaccine strain (IND 17/82), warranting the selection of a suitable vaccine strain which can cover this diversity in antigenic spectrum. In earlier studies employing neutralization test with anti-146S rabbit sera raised against eight candidate vaccine strains, IND 81/00 and IND 40/00 belonging to genotype VII were found to offer the best antigenic coverage. In order to assess the credibility of IND 81/00 and IND 40/00 as vaccine strains, 17 recent isolates received during 2005-2006 and representative isolates from older genotypes were subjected to two-dimensional micro-neutralization assay using bovine convalescent serum (against IND 81/00 and IND 40/00) and bovine vaccinate serum (against IND 40/00). From the results it is evident that both the isolates IND 81/00 (antigenic relationship 'r-value' >0.40 with 86% of isolates) and IND 40/00 ('r-value' >0.40 with 78% of isolates) show nearly equal antigenic relatedness with the recent field viruses and hence both of these are effective vaccine candidates in present context. Though very limited in its extent, these useful data obtained with antisera raised in homologous host system are logical extension of the on going quest for the appropriate vaccine strain and circumvents species disparities in the immune recognition of epitopes. PMID- 18394828 TI - Oxygen tension and medium type actions on blastocyst development and interferon tau secretion in cattle. AB - Most current in vitro production systems terminate at the blastocyst stage in cattle. The goal of the present research was to identify culture conditions that support individual blastocyst survival and interferon-tau (IFNT) production in cattle. In the first study, two media (medium 199 [M199] and potassium simplex optimized medium [KSOM]) and two oxygen tensions (5 and 20%) were compared for their ability to sustain blastocyst survival and IFNT production from days 8 to 11 post-insemination. Survival and total cell numbers were greater (P<0.05) for blastocysts cultured in M199 in a 5% oxygen environment compared with other medium and oxygen treatment combinations. Serum supplementation was required for blastocyst survival and IFNT production. IFNT concentrations in conditioned medium were similar for blastocysts cultured in M199 or KSOM, but blastocysts incubated in 5% oxygen produced less (P<0.001) IFNT than their 20% oxygen counterparts. Oxidative stress was not responsible for the increase in IFNT concentrations. Supplementation with fibroblast growth factor 2 did not affect cell numbers but increased (P<0.02) IFNT concentrations for blastocysts cultured in 5% oxygen but not those cultured in 20% oxygen. In conclusion, culturing blastocysts of cattle in a 5% oxygen environment with M199 containing serum sustains embryo viability and permits constitutive and inducible IFNT production. Incubation in 20% oxygen increases IFNT production. The mechanism responsible for this event and its physiological relevance to conceptus development in utero remain unknown. PMID- 18394829 TI - One-year therapy with 10mg/day DHEA alone or in combination with HRT in postmenopausal women: effects on hormonal milieu. AB - The purpose of this study was to evaluate the effects on hormonal milieu of 1 year therapy with 10 mg/day oral dehydroepiandrosterone (DHEA) or 50 microg transdermal estradiol plus 100 mg/day oral micronized progesterone in a group of 20 healthy postmenopausal women (age=50-58 and years since menopause (ysm)=1-6) and also the effects observed by combining these two therapies in a group of 12 postmenopausal women (age=54-61 and ysm=6-10) characterized by lower baseline DHEA and DHEAS levels (<2.40 and <0.55 microg/ml, respectively). DHEA produced a significant rise in androgens levels, whereas HRT did not. Moreover, DHEA alone induced a significantly lower increase in estrogens and beta-endorphin levels and a higher decrease in cortisol levels than HRT. DHEA and HRT also produced a significant similar increase in allopregnanolone levels. DHEA plus HRT induced a significantly higher increase in testosterone and estradiol and a lower increase in allopregnanolone and beta-endorphin levels and a significantly lower decrease in cortisol levels than HRT alone treated group. A similar increase was observed in progesterone and SHBG levels in all groups. These results suggest that 10-mg DHEA seems to be the proper dose to replace androgen deficiency in subjects with reduced Delta-5 androgens plasma levels. However, the aging process and the number of years since menopause may further modulate the effects of hormone therapy on hormonal milieu. PMID- 18394830 TI - Stability of lyophilized human growth hormone. AB - To evaluate relationships between the extent of protein-excipient interactions, structural relaxation of an amorphous matrix, and the physico-chemical stability of a protein, human growth hormone (hGH) was lyophilized with sucrose and trehalose in a 1:2 weight ratio. The protein-excipient interactions were analyzed immediately after lyophilization with isoperibol solution calorimetry (ISC), water sorption analysis (WSA), differential scanning calorimetry (DSC) and Fourier Transform Infrared Spectroscopy (FTIR). The physical and chemical stability of hGH during storage at 50 degrees C was monitored by reverse phase (RP)-HPLC, SEC-HPLC and UV absorption spectroscopy. The hGH formulation containing sucrose demonstrated greater protein-excipient interactions and faster initial relaxation times compared to the trehalose formulation. Although both formulations had similar chemical stability (rate of deamidation), physical stabilities (e.g. degree of aggregation) were different. The hGH/sucrose formulation manifested a higher rate and lower extent of insoluble aggregate formation. The decreased amount of aggregation in the sucrose formulation could be correlated with a greater extent of protein-excipient interactions and the presence of a more homogeneous mixture. In contrast, the higher rate of aggregation in the sucrose formulation could be directly correlated with the higher molecular mobility of the matrix. PMID- 18394831 TI - Application of process analytical technology in tablet process development using NIR spectroscopy: blend uniformity, content uniformity and coating thickness measurements. AB - Near-infrared (NIR) spectroscopy was employed as a process analytical technique in three steps of tabletting process: to monitor the blend homogeneity, evaluate the content uniformity of tablets and determine the tablets coating thickness. A diode-array spectrometer mounted on a lab blender (SP15 NIR lab blender) was used to monitor blend uniformity using a calibration-free model with drug concentration ranging from 2.98 to 9.25% (w/w). The method developed accurately depicted the changes in concentration of the drug during blending and the positive effect of a delumping step in the production process. Blend homogeneity was reached within 2 min of the blending step post-delumping, with relative standard deviation (R.S.D.) values varying from 1.0 to 2.5% depending on the drug concentration of the blend. A Fourier-transform spectrometer (Bruker MPA) was used to analyze content uniformity and coating thickness with calibration based models. Prediction of a validation set with tablets compacted at pressures not present in the calibration set yielded an root mean square error of cross validation (RMSEP) of 1.94%; prediction of tablets compacted at pressures present in the calibration set yielded a RMSEP of 1.48%. Performance of the model was influenced by several physical tablet properties, which could be reduced by spectral pre-processing. A model based on reflectance spectra predicted coating thickness and its variation more accurately than the model based on transmission spectra. Inter-tablet coating variation was predicted with NIR and compared to reference thickness measurements. Both methods gave comparable results. Initial inter-tablet variation of tablets sampled in-process during coating was high, but stabilized after 30 min into the process. PMID- 18394832 TI - Efficacy and toxicity of Eudragit-coated chitosan-succinyl-prednisolone conjugate microspheres using rats with 2,4,6-trinitrobenzenesulfonic acid-induced colitis. AB - A targeted delivery system for inflammatory bowel disease (IBD), Eudragit L100 (EuL)-coated chitosan (Ch)-succinyl-prednisolone (SP) conjugate microspheres (Ch SP-MS/EuL), were designed and examined in vivo for efficacy and toxicity. Their preparation was conducted in the same manner as previously; that is, by synthesis of the conjugate by carbodiimide coupling of Ch and SP, conversion into microspheres (Ch-SP-MS), and coating of Ch-SP-MS with EuL. Experimental colitis was induced by instillation of 2,4,6-trinitrobenzenesulfonic acid (TNBS) into the colon in rats. Drugs were administered once or twice a day intragastrically for three consecutive days. Visible colitis severity, colon/body weight ratio and myeloperoxidase activity were measured as inflammatory indices. Toxicity was examined from the decrease in the thymus/body weight ratio. Efficacy was dose dependent and the greatest in the order Ch-SP-MS/EuL>Ch-SP-MS>prednisolone (PD) alone, and Ch-SP-MS/EuL showed excellent recovery of colitis states. Toxicity was the greatest in the order PD>>Ch-SP-MS>Ch-SP-MS/EuL. Ch-SP-MS and Ch-SP-MS/EuL reduced significantly the thymic atrophy caused by PD. It was demonstrated that Ch-SP-MS/EuL enhanced effectiveness of PD and reduced toxic side effects of PD greatly. Also, these results established the prediction by previous in vitro and in vivo studies. PMID- 18394833 TI - Boronate-containing polymers form affinity complexes with mucin and enable tight and reversible occlusion of mucosal lumen by poly(vinyl alcohol) gel. AB - Copolymers of N-acryloyl-m-aminophenylboronic acid (NAAPBA) with N,N dimethylacrylamide (DMAA) formed insoluble interpolymer complexes with mucin from porcine stomach at pH 9.0. The complex formation based on boronate-sugar interactions took place between the similarly charged macromolecules and resulted in coacervate particles formation, which depended both on pH and ionic strength of the solution. The coacervation rate displayed a maximum at the intermediate DMAA-NAAPBA copolymer: mucin weight ratio, that is a pattern typical of interpolymer complex formation. The effective hydrodynamic particle diameter of the coacervates monotonously grew from 155+/-20 nm up to 730+/-120 nm in 2 days in 0.1M sodium bicarbonate buffer solution, pH 9.0. Electrophoretic mobility of the resultant nanoparticles was intermediate between those of individual polymers, whereas the particles zeta-potential was -7.5+/-0.4 mV in the above buffer solution. Pre-treatment of the inner mucosal epithelium of excised male pig urethras with 5% (w/v) solutions of acrylamide-NAAPBA or DMAA-NAAPBA copolymers at pH 8.8 allowed for tight occlusion of the lumen by poly(vinyl alcohol)-borax gel injected via a two-way catheter. Leakage of 0.15M NaCl solution through the thus occluded organs could be prevented, while the leakage through the organs occluded by the gel without the pre-treatment was unavoidable. The gel plug could be quickly dissolved on demand after injection of 5% (w/v) aqueous fructose solution into the lumen. The described technique may be useful for temporal occlusion of mucosal lumens in living organisms. In contrast to the conventional mucoadhesive polymers like polyacrylic acid or chitosan, the boronate-containing copolymers display their mucoadhesivity at weakly alkaline pH of 8-9 and physiological ionic strength. PMID- 18394834 TI - Relationship between dissolution efficiency of Oxazepam/carrier blends and drug and carrier molecular descriptors using multivariate regression analysis. AB - Quantitative structure-property relationships were developed for predicting the enhancement of dissolution rate of the model lipophilic drug Oxazepam (Oxa) from blends (BLs) with 12 structurally different carriers at three different drug/carrier weight ratios (1/5, 1/10, and 1/20). To this end, 36 BLs were prepared by the solvent-evaporation method and characterized by spectroscopic (FT IR), thermoanalytical (DSC) and X-ray diffraction studies. The dissolution rate of the examined systems was quantified by logDE/DE(Oxa), where DE and DE(Oxa) are the dissolution efficiencies of the BL and pure drug, respectively. Twenty molecular descriptors, including parameters for size, lipophilicity, cohesive energy density (CED), and hydrogen bonding capacity were calculated and together with the experimental melting point (MP), were used in multivariate analysis. Twelve pertinent variables were detected after looking at the results of principal component analysis (PCA) and cluster analysis, and reliable six descriptor models generated by Partial Least Squares-Projection to Latent Structures (PLS) method. Satisfactory coefficient of determination values were obtained (i.e., R(2) equal to 0.794 and Q(2) equal to 0.705). The equations generated can predict with reasonable accuracy the dissolution rate increase of the model lipophilic drug/carrier BLs. PMID- 18394835 TI - In vitro antifungal activity of extracts and neolignans from Piper regnellii against dermatophytes. AB - The present study was designated to evaluate the in vitro antidermatophyte activity of extracts from leaves of Piper regnellii as well as of the bioactivity directed isolation of neolignans. The antifungal assay was performed by microdilution techniques. The hydroalcoholic extract of Piper regnellii leaves presented a strong activity against the dermatophyte fungi Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis and Microsporum gypseum with MICs of 15.62, 15.62, 15.62 and 62.5 microg/ml, respectively. On light microscopy and scanning electron microscopy of nail fragments not exposed to hydroalcoholic extract of Piper regnelli leaves, well-formed and extensive mycelial growth was seen. On nail fragments exposed to hydroalcoholic extract at concentrations more than 1.2mg/ml and then inoculated with spore suspension, growth was not seen. The hydroalcoholic extract was fractionated on silica gel in to nine fractions. The active chloroform fraction was lyophilized and chromatographed by column chromatography on silica gel. Structures were established by comparison with literature data and identified as eupomatenoid-3 and eupomatenoid-5. The pure compounds showed strong activity on Trichophyton rubrum with MIC of 50 and 6.2 microg/ml, respectively. Comparing the activity of the active chloroform fraction obtained from hydroalcoholic crude extract with that of isolated compound eupomatenoid-5, it is clear that this showed the same results against Trichophyton rubrum. The results showed that the plant could be explored for possible antifungal agents and provides preliminary scientific validation for the traditional medicinal use of this plant. PMID- 18394836 TI - Ion channel mechanism and ingredient bases of Shenfu Decoction's cardiac electrophysiological effects. AB - AIM OF THE STUDY: Ion channel mechanism of cardiac electrophysiological effects of Shenfu Decoction (SFD, Ginseng and Aconiti Praeparatae Decoction), a traditional Chinese medicine (TCM) prescription, and its ingredient bases were investigated in guinea pigs. MATERIALS AND METHODS: After administration of an injection made from SFD (Shenfu Injection, SFI), the indexes of transmembrane action potential (TAP) in vivo and sodium channels in isolated ventricular myocyte were assayed by suspended microelectrodes and patch clamp techniques respectively, and ingredients of SFD were compared with. RESULTS: After administration of SFI, the action potential amplitude (APA) and maximum velocity (V(max)) of TAP decreased. In the presence of either SFI or Fuzi active ingredient (FZAI)(5, 10, and 15%), not any other ingredient, the density of voltage-dependent sodium current (I(Na)) decreased significantly, while the inhibition ratio of SFI was larger. EC(50) of SFI was less than the one of FZAI, and SFI displayed effects on I(Na) in wider voltage scope than FZAI in current voltage curve. Both SFI and FZAI shifted the steady-state inactivation curve of sodium channels to the left, and the recovery curve to the right. CONCLUSIONS: The results indicated that the cardiac electrophysiological effects of SFI were exerted by blocking sodium channels, and FZAI contributed to such effects most but inferior to SFI, which justified its use in anti-arrhythmia, myocardial protection, etc. PMID- 18394837 TI - [Various typing methods of Pseudomonas aeruginosa strains isolated from cystic fibrosis patients]. AB - Typing methods are essential to understand the epidemiology of bacterial infections. Strain typing is important for the detection of sources or routes of infections, identification between endemic and epidemic strains and prevention of transmission between patients. Some Pseudomonas aeruginosa cystic-fibrosis strains could not be typed with conventional typing methods. Due to the diverse phenotypic nature of P. aeruginosa, phenotyping methods are not discriminatory enough to identify strains belonging to the same genotype. Thus, molecular typing methods are required. These methods should be applied when data from phenotypic typing analysis becomes ambiguous, such as in cystic fibrosis. Molecular typing methods, developed over the past decade, are highly discriminatory in capacity and reproducibility. However, they are more likely applied in specialized laboratories since they are expensive and increase the workload. A reliable and low-cost typing system is required for better defining the epidemiology of this pathogen and designing more rational policies of infection control. Comparison between typing methods will pinpoint the limits and effectiveness of each method and will improve in turn the choice of a nonspecialized laboratory in terms of simplicity, time and cost. PMID- 18394838 TI - [Severe lower gastrointestinal bleeding related to prostate biopsy]. PMID- 18394839 TI - Secretory diarrhoea with high faecal potassium concentrations: a new mechanism of diarrhoea associated with colonic pseudo-obstruction? Report of five patients. AB - OBJECTIVE: To report the mechanism of diarrhoea in patients with subacute colonic pseudo-obstruction, profuse secretory diarrhoea and hypokalemia. PATIENTS: Five consecutive patients who developed colonic pseudo-obstruction, profuse watery diarrhoea and severe hypokalemia. Investigations excluded mechanical intestinal obstruction. Usual cause of diarrhoea were ruled out. Abdominal distension and diarrhoea improved simultaneously in all cases after colonoscopic decompression or intravenous neostigmine. RESULTS: Faecal ionograms showed a low osmotic gap and high faecal potassium concentration explaining the hypokalemia: 100 to 180 mEq/kg (usually inferior than 50 mEq/l in case of secretory diarrhoea) and low faecal sodium concentrations. Potassium salts were the only factor identified as the driving osmotic force for the diarrhoea. CONCLUSION: Secretory diarrhoea is classically due to chloride active secretion with passive sodium secretion or to inhibition of sodium absorption. In five cases of Ogilvie's syndrome we evidenced an original mechanism of secretory diarrhoea due to active potassium secretion responsible of a profound hypokalemia. This novel type of diarrhoea may be a hallmark of colonic pseudo-obstruction due to colonic distension. PMID- 18394840 TI - Identification of a degradation product in stressed tablets of olmesartan medoxomil by the complementary use of HPLC hyphenated techniques. AB - An unknown degradation product (DP-1) increased in olmesartan medoxomil (OLM) tablets stored at 40 degrees C/75% r.h., reaching 0.72% after 6 months. The molecular weight and fragment information obtained by LC-MS suggested that DP-1 was a dehydrated dimer of olmesartan (OL) and the presence of ester carbonyl group was indicated by solvent-elimination LC-IR analysis. LC-(1)H NMR confirmed the structure of DP-1 as an esterified dimer of OL. Rapid and accurate identification of the degradation product was achieved by the complementary use of HPLC hyphenated techniques without complicated isolation or purification processes. PMID- 18394842 TI - Quantitative determination of underivatized polyamines by using isotope dilution RP-LC-ESI-MS/MS. AB - A rapid, sensitive and selective method using LC-MS/MS was developed and validated for the simultaneous quantitative determination of five polyamines N1,N12-diethylspermine (DESpm), N-ethylspermine (EtSpm), N1-ethylspermidine (EtSpd), spermidine (Spd) and N1-ethyl-1,3-diaminopropane (EtDAP) without any derivatization steps. The LC-MS/MS system was operated using the positive electrospray ionization (ESI) mode. The chromatographic separation only took 10min and was performed on a reversed phase C18 column with 0.1% heptafluorobutyric acid as the ion-pairing agent and acetonitrile gradient. Stable, deuterium labelled internal reference compounds of the five analytes were included in the quantification. The lower limit of quantification for all of the five analytes was 0.03microM and the method was linear for DESpm, EtSpd, Spd and EtDAP over the range of 0.03-60microM and for EtSpm over the range of 0.03 30microM. Correlation coefficients (R2) were always >0.995 for all the analytes. The precision of the overall method ranged from 0.2 to 9.7% as intra-day variability and from 0.9 to 6.8% as inter-day variability. The intra-day and inter-day accuracy of the assay ranged between 87.6-109.8% and 89.6-106.6%, respectively. The method has been applied successfully to quantify metabolites of DESpm as a substrate for recombinant human polyamine oxidase. PMID- 18394841 TI - Characterization of phenolic compounds in the Chinese herbal drug Artemisia annua by liquid chromatography coupled to electrospray ionization mass spectrometry. AB - A simple and rapid method has been established for the screening of the main phenolic compounds in Artemisia annua by LC-DAD-ESI-MS(n). A total of 40 phenolic compounds were identified or tentatively characterized in the methanol extract of A. annua, including 8 C-glycosyl flavonoids, 5 O-glycosyl flavonoids, 3 flavonoid aglycones, 21 quinic acid derivatives, 2 benzoicacid glucosides and 1 coumarin. The C-glycosyl flavonoids were reported from A. annua for the first time and they were found to be a new type of main constituents, and might be responsible for its antioxidant and antiviral activity. Quinic acid derivatives were also found to be the major constituents of A. annua. PMID- 18394843 TI - Quantitative analysis of cocaine in human hair by HPLC with fluorescence detection. AB - Cocaine is currently one of the most widespread abuse drugs in the world. Since hair cocaine concentrations are a reliable marker of exposition to the drug, an original liquid chromatographic method has been developed for the determination of cocaine in human hair. The chromatographic analysis was carried out on a Hydro RP C18 column, using a mobile phase containing a phosphate buffer (pH 3.0) acetonitrile-methanol (75:15:10, v/v/v). Native cocaine fluorescence was monitored at 315nm while exciting at 230nm. Mirtazapine was used as the internal standard. Sample pre-treatment was carried out by incubative extraction with 0.1M HCl followed by solid-phase extraction with C2 cartridges. Good linearity was obtained over a working range of 0.3-100.0ng/mg. Both extraction yield (>89%) and precision values (R.S.D.<6.2%) were highly satisfactory. The method was successfully applied to hair samples collected from cocaine users. Thus, the method is suitable for the long-term monitoring of cocaine use by means of hair testing. PMID- 18394844 TI - Development of multiplex real-time hybridization probe reverse transcriptase polymerase chain reaction for specific detection and differentiation of Enterovirus 71 and Coxsackievirus A16. AB - Large outbreaks of hand, foot, and mouth disease have been reported in the Asia Pacific region over the last few years and resulted in significant fatalities. The 2 main etiologic agents are Enterovirus 71 (EV71) and Coxsackievirus A16 (CA16). Both viruses are closely related genetically and show similar clinical symptoms. However, EV71 are associated with neurologic complications and can lead to fatalities. In this study, we developed a multiplex real-time hybridization probe reverse transcriptase polymerase chain reaction to detect and differentiate EV71 from CA16 using the LightCycler (Roche Molecular Biochemicals). Specific primers and hybridization probes were designed based on highly conserved VP1 region of EV71 or CA16. Our results showed high specificity and sensitivities in detecting EV71 or CA16 from 67 clinical specimens, and no other enterovirus serotype was detected. Rapid diagnosis to differentiate EV71 from CA16 in outbreak situations will enable pediatricians to identify and manage the patients more effectively. PMID- 18394845 TI - Characterization of ST80 Panton-Valentine leukocidin-positive community-acquired methicillin-resistant Staphylococcus aureus clone in Tunisia. AB - The spread of community-acquired methicillin-resistant Staphylococcus aureus (CA MRSA) has been reported in communities worldwide. In this study, we characterized 64 Tunisian CA-MRSA by agr typing, polymerase chain reaction assay for 20 virulence genes, staphylococcal chromosomal cassette mec (SCCmec) typing, pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and protein A gene (spa) typing. All our isolates were lukS-PV-lukF-PV positive, etd positive, and edin positive. They harbored SCCmec type IV and belonged to agr group 3. PFGE typing showed that our isolates were distributed in 11 different pulsotypes. spa typing and MLST, performed with isolates representative of each PFGE pattern, revealed that all isolates had a unique spa type (t044) and a common sequence type (ST80). The isolates showed susceptibility to the majority of antibiotics, and resistance to kanamycin, erythromycin, and tetracycline, but intermediate resistance to fusidic acid. Full analysis of our results revealed that our isolates were nonmultiresistant and belonged to a single clonal type ST80. PMID- 18394846 TI - Primitive transport vehicle (varangele) accidents. PMID- 18394847 TI - Repeat Wellens' syndrome: case report of critical proximal left anterior descending artery restenosis. AB - We describe the case of a 40-year-old woman who presented to the Emergency Department with resolving chest pain. The initial electrocardiogram (ECG) showed biphasic T-waves in V2-V4, which was recognized as Wellens' syndrome, or acute coronary T-wave syndrome. Emergent cardiac catheterization revealed 95% stenosis of the previously placed stent in the proximal left anterior descending artery (LAD). Review of her records from 3 years prior revealed similar ECG findings consistent with Wellens' syndrome, at which time a stent was placed for critical proximal LAD stenosis. We report this case to increase awareness of the T-wave abnormalities associated with Wellens' syndrome. There is significant morbidity and mortality that can occur in the absence of emergent coronary revascularization. This report of repeated Wellens' syndrome in the same patient demonstrates both types of precordial T-wave abnormalities that characterize Wellens' syndrome. PMID- 18394848 TI - Sexual assault victims in the emergency department: analysis by demographic and event characteristics. AB - The objective of this study was to analyze demographic and event characteristics of patients presenting to the Emergency Department (ED) for evaluation after sexual assault, using a Sexual Assault Nurse Examiner standardized database. Data were prospectively collected as part of the Sexual Assault Nurse Examiner program at an urban teaching hospital. This study reviewed all ED patient records with a complaint of sexual assault between January 1, 2000 and December 31, 2004. Data were collected on 1172 patients; 92.6% were women, with a mean age of 27 years. The sample was 59.1% black, 38.6% white, and 2.3% "Other." Black victims of sexual assault were significantly more likely to be young (25 years or less) than Whites. Over half (54%) reported involvement of drugs or alcohol during the event. Fifty-three percent knew their assailant(s), and black and young patients were significantly more likely to know the perpetrator(s). Threats of force were common (72.4% of sample), and multiple assailants were uncommon (18.1% of sample). Physical evidence of trauma was present in more than half (51.7%), with increased rates among Whites and older persons. Multivariate analysis showed that race, age, threats, and substance use during the event were independent risk factors for evident trauma on physical examination. Survivors of sexual assault who present to the ED are overwhelmingly female, relatively young, often know the perpetrator of the event, and are likely to be threatened and show signs of physical trauma. Differences between patients according to demographic and event characteristics may have important implications for ED management and treatment plans. PMID- 18394849 TI - Subcapsular renal hematoma complicating acute pyelonephritis. AB - Subcapsular renal hematoma (SRH) is an extremely rare complication of acute pyelonephritis (APN). We report a case of SRH complicating APN with a ureteral stone, together with a review of the literature. PMID- 18394850 TI - Gastric rupture from cardiopulmonary resuscitation or seizure activity? A case report. AB - Ascertaining the cause of a gastric rupture is difficult and can be complicated by events occurring in the pre-hospital phase of patient care. To our knowledge, only seven cases of cardiopulmonary resuscitation (CPR)-related injuries to the stomach have been reported in the literature since 1997. We found no reports of seizure-related gastric injury. We present the case of a patient who sustained a gastric rupture, tension pneumoperitoneum, and a tension pneumomediastinum after retching, seizure, CPR, and advanced cardiac life support maneuvers. PMID- 18394851 TI - Bradycardia and confusion. PMID- 18394852 TI - Lupus anticoagulant syndrome: a case report. AB - Thromboses of the deep veins (DVT) and potential complications are disorders with which physicians need to be well familiarized so they can be recognized and managed. Hypercoagulable states play a significant role in the development of DVT and these disorders must be suspect when thrombosis occurs, especially in the upper extremities. Antiphospholipid antibody syndrome (APS) is one such hypercoagulable state in which autoantibodies are formed against anionic phospholipid complexes. In vivo, these antibodies cause a hypercoagulable state through a number of proposed mechanisms, whereas in vitro they interfere with the assembly of phospholipid complexes, thereby inhibiting coagulation and prolonging various clotting times. Though appearing to be anticoagulated on ancillary testing, patients with APS are actually in a pro-thrombotic state requiring treatment with the anticoagulants heparin and warfarin. This case discussion focuses on the recognition and treatment of thrombotic events in the setting of APS. PMID- 18394853 TI - The (f)utility of bedside registration? PMID- 18394854 TI - Phantom shocks in patients with an implantable cardioverter defibrillator. AB - Phantom shock is the sensation of shock in the absence of an actual implantable cardioverter-defibrillator (ICD) discharge. The ICD is now the first-line therapy for patients with ventricular tachycardia and fibrillation. There has been a significant increase in the number of patients with an ICD and patients presenting to the Emergency Department (ED) after a shock for evaluation and device interrogation. Phantom shock is more likely to be nocturnal in the first 6 months after implantation, and patients are more likely to be clinically depressed and have higher levels of anxiety. There is no specific treatment. We report 3 patients who presented to the ED with the sensation of ICD discharges, however, on device interrogation had no shocks and no dysrhythmias. PMID- 18394855 TI - Gas1 reduces Ret tyrosine 1062 phosphorylation and alters GDNF-mediated intracellular signaling. AB - The present results show that the expression of Growth Arrest Specific1 (Gas1) in SH-SY5Y neuroblastoma cells significantly inhibits the increased phosphorylation of tyrosine 1062 of the Ret receptor tyrosine kinase induced by glial-cell-line derived neurotrophic factor (GDNF). We also observed that Gas1 significantly reduces the activation of Akt. GDNF and members of its family of ligands (GFLs), signal through a molecular complex consisting of one of its receptors (GFRalphas) and the Ret receptor tyrosine kinase. GDNF is a key component to preserve several cell populations in the nervous system, including dopaminergic and motor neurons, and also participates in the survival and differentiation of peripheral neurons such as enteric, sympathetic and parasympathetic. On the other hand, Gas1 is a molecule involved in cell arrest that can induce apoptosis when over-expressed in different cell lines, including cells of neuronal and glial origin. Although, Gas1 is widely expressed during development, its role in vivo has not yet been clearly defined. We recently showed the structural homology between Gas1 and GFRalphas, thus suggesting that the physiological role of Gas1 is that of modulating the biological responses induced by GDNF and/or other members of this family of signaling molecules. The results of this work are consistent with the hypothesis of Gas1 acting as a negative modulator of GDNF signaling. PMID- 18394856 TI - A case for clarity in the writing of health statements. AB - OBJECTIVE: While effectively written health-related materials can facilitate self care, using negative wording may backfire, as patients may forget negations and remember the opposite of intended information. The present study attempts to determine if older readers are particularly susceptible to misremembering negatively worded health-related statements. METHODS: Younger and older adults read positively and negatively worded statements and were later tested for memory of the exact statements, statements opposite in meaning due to the addition or deletion of a negation, and entirely new statements. RESULTS: For intact statements, both groups recognized more positive than negative statements. When statements reversed meanings, younger adults were more likely to accurately recognize items that changed from negative to positive than were older adults, who were more likely to incorrectly endorse changed items as intact when items began negative and became positive than the reverse. CONCLUSION: The inclusion of negations, often used in attention-grabbing techniques such as myth-busting, can backfire, especially among older audiences who are less likely to accurately remember negatively worded items and especially likely to endorse their positive counterparts. PRACTICE IMPLICATIONS: Designers of healthcare materials should avoid using negatively worded statements, which may be misremembered by vulnerable populations, in favor of positive wording. PMID- 18394857 TI - Expanding use of emergency endovascular repair for ruptured abdominal aortic aneurysms: disparities in outcomes from a nationwide perspective. AB - BACKGROUND: Endovascular repair (EVAR) of abdominal aortic aneurysms (AAA) has become widely accepted in the elective setting but remains controversial for emergency repair of ruptured aneurysms (rAAA). We sought to examine the national trends in use and associated outcomes with EVAR. METHODS: The Nationwide Inpatient Sample (NIS) was used to analyze all admissions for rAAA from 2001 through 2004. Nationwide temporal trends and demographics using weighted samples were evaluated. Focused univariate and multivariate analyses comparing outcomes from open repair and EVAR were done for the years 2003 and 2004. RESULTS: There were 28,123 admissions for rAAA, with a stepwise decline in admissions from 2001 to 2004. Use of EVAR increased significantly from 6% of all emergency repairs in 2001 to 11% in 2004 (P < .01). Mortality for EVAR declined significantly from 43% to 29% (P < .01), but mortality with open repair showed no change (40% to 43%). From the 2003 to 2004 data set, 949 EVAR and 8982 open repairs were identified. Compared with open repair, the EVAR patients had lower mortality (31% vs 42%), shorter hospital stay (6 vs 9 days), and were more likely to be discharged to home (59% vs 37%, all P < .01). The total hospital charges for EVAR and open repair were similar (dollars 71,428 vs $74,520, P = .59). Mortality for EVAR was significantly higher at nonteaching hospitals compared with teaching centers (55% vs 21%, P < .01) and at nonteaching centers, even exceeding that of open repair (46%). Regression modeling confirmed the overall benefits of EVAR as well as the worse outcomes at nonteaching facilities after adjusting for patient comorbidities, disease severity, and hospital or system covariates. CONCLUSIONS: Endovascular repair is being increasingly used in the emergency management of ruptured AAA, with steadily decreasing mortality during the study period. Endovascular AAA repair is associated with improved mortality and outcomes compared with open repair, but results in nonteaching centers are substantially worse than those in teaching hospitals. PMID- 18394858 TI - [Managing comorbidities in hepatology: toward a more holistic medicine]. AB - Approximately 20% of patients infected with the hepatitis B or C virus (HBV and HCV) develop cirrhosis of the liver. It is essential, especially for preventive purposes, to test for related etiological factors, especially excess alcohol consumption, metabolic syndrome, and obesity. The frequency of these health problems and their hepatic tropism explain these frequent associations. In patients with chronic HBV and HCV, alcohol consumption and metabolic syndrome increase the risk of fibrosis; in those with HCV, they also reduce the likelihood of treatment response. In patients with alcoholic hepatitis, overweight increases cirrhotic risk. If cytolysis persists after the identified factor is controlled, another etiologic factor must be sought and treated. For patients with excess alcohol consumption and similarly those with metabolic syndrome, it is essential to differentiate between dependency, which is more difficult to treat, and other risk situations, for which the efficacy of a brief intervention by the physician has been demonstrated. In this more holistic approach, the physician treats a person with liver disease, rather than just a diseased liver. PMID- 18394859 TI - Absence of cognitive deficits following deep brain stimulation of the subthalamic nucleus for the treatment of Parkinson's disease. AB - Electrical stimulation of the subthalamic nucleus is an effective treatment for the motor symptoms of Parkinson's disease. While most patients who undergo this procedure do not appear to suffer behavioral side effects, a minority experience cognitive or emotional deficits, and longitudinal studies have reported declines; however, the measures of cognitive function used have been limited. One explanation for the possible disturbance of cognitive functions is that electrical stimulation of the subthalamic nucleus disrupts the normal flow of information within cortico-striatal loops involving prefrontal, associative, or limbic cortex. We wished to assess the effect of high frequency electrical stimulation of the subthalamic nucleus in Parkinson's disease patients while they performed a comprehensive neuropsychological test battery. We selected cognitive tasks known to test the function of different cortical areas, including tests of executive function, cognitive flexibility, attention, memory, language and visual perception. Patients were tested on two separate days, with the stimulators turned on or off. Test scores were also compared to preoperative performance. In our sample of 15 patients without dementia or major depression there was no deterioration on any cognitive test as a result of stimulation. We conclude that electrical stimulation of the motor subthalamic nucleus does not cause appreciable declines in cognitive function in well-selected patients. PMID- 18394860 TI - Predicting children's post-traumatic stress symptoms following hospitalization for accidental injury: combining the Child Trauma Screening Questionnaire and heart rate. AB - This study investigated the utility of combining the Child Trauma Screening Questionnaire (CTSQ) [Kenardy, J. A., Spence, S. H., & Macleod, A. C. (2006). Screening for post-traumatic stress disorder in children after accidental injury. Pediatrics, 118, 1002-1009] and children's heart rate (HR; emergency department and 24-h post-admission) to identify children likely to develop post-traumatic stress disorder (PTSD) symptoms at 1 and 6 months post-injury. Children completed the CTSQ within 2 weeks of injury. PTSD symptoms were assessed with the Anxiety Disorders Interview Schedule for DSM-IV [Silverman, W. K., & Albano, A. M. (1996). Anxiety Disorders Interview Schedule for DSM-IV, Child Version, Parent Interview Schedule. Orlando, Florida: The Psychological Corporation], for 79 children aged 7-16 years. A combination of the CTSQ plus HR (CTSQ-HR) was better than the CTSQ alone or HR alone at identifying children likely to develop PTSD symptoms. These findings suggest that the CTSQ-HR screen may increase identification of children who are likely to develop PTSD symptoms, enabling development of targeted prevention programs. PMID- 18394862 TI - Investigations of the effects of the antimalarial drug dihydroartemisinin (DHA) using the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). AB - Artemisinin derivatives are effective and safe drugs for treating malaria, but they are not recommended during the first trimester of pregnancy because of resorptions and abnormalities observed in animal reproduction studies. Previous studies in rats showed that artemisinin embryotoxicity derives from the depletion of primitive red blood cells (RBCs) over a narrow critical time window (gestation Days 9-14). In order to further investigate the susceptibility of primitive RBCs to artemisinins and to establish whether this susceptibility is species-specific or inherent to the compound, we studied dihydroartemisinin (DHA), both a drug in its own right and the main metabolite of current artemisinin derivatives in use, in the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). This model readily allows investigation and monitoring of primitive and definitive RBCs. Effects on frog larvae exposed to DHA for 48 h during early embryonic development, starting from 24 h post fertilization, were similar to those on rat embryos in terms of reduction in the number of primitive RBCs (clonally produced within the ventral blood island). In contrast, RBCs of older larvae (stage 47, produced at the definitive sites of hematopoiesis) were affected minimally and subsequently recovered. Compared to rat embryos, the frog larvae had no areas of necrosis but they shared similar heart defects. The mitochondrion appeared to be the main subcellular target, similar to observations in Plasmodium. These results implicate artemisinin-induced embryotoxicity through perturbation of metabolically active RBCs; whereas this mode of action does not appear to be species-specific, the stages of susceptibility varied between different species. The window of susceptibility and duration of exposure must be considered to evaluate the clinical relevance of these findings. PMID- 18394863 TI - To dichotomize or not to dichotomize? PMID- 18394861 TI - Social regulation of leukocyte homeostasis: the role of glucocorticoid sensitivity. AB - Recent small-scale genomics analyses suggest that physiologic regulation of pro inflammatory gene expression by endogenous glucocorticoids may be compromised in individuals who experience chronic social isolation. The present study assessed the relationship between leukocyte distributional sensitivity to glucocorticoid regulation and subjective social isolation in a large population-based sample of older adults. Initial analyses confirmed that circulating neutrophil percentages were elevated, and circulating lymphocyte and monocyte percentages were suppressed, in direct proportion to circulating cortisol levels. However, leukocyte distributional sensitivity to endogenous glucocorticoids was abrogated in individuals reporting either occasional or frequent experiences of subjective social isolation. This finding held in both non-parametric univariate analyses and in multivariate linear models controlling for a variety of biological, social, behavioral, and psychological confounders. The present results suggest that social factors may alter immune cell sensitivity to physiologic regulation by the hypothalamic-pituitary-adrenal axis in ways that could ultimately contribute to the increased physical health risks associated with social isolation. PMID- 18394864 TI - NKCC1 and KCC2 prevent hyperexcitability in the mouse hippocampus. AB - During postnatal development of the central nervous system (CNS), the response of GABA(A) receptors to its agonist undergoes maturation from depolarizing to hyperpolarizing. This switch in polarity is due to the developmental decrease of the intracellular Cl concentration in neurons. Here we show that absence of NKCC1 in P9-P13 CA3 pyramidal neurons, through genetic manipulation or through bumetanide inhibition, results in a significant increase in cell excitability. Furthermore, the pro-convulsant agent 4-aminopyridine induces seizure-like events in NKCC1-null mice but not in wild-type mice. Measurements of muscimol responses in the presence and absence of NKCC1 shows that the Na-K-2Cl cotransporter only marginally affects intracellular Cl(-) in P9-P13 CA3 principal neurons. However, large increases in intracellular Cl(-) are observed in CA3 pyramidal neurons following increased hyperexcitability, indicating that P9-P13 CA3 pyramidal neurons lack robust mechanisms to regulate intracellular Cl(-) during high synaptic activity. This increase in the Cl(-) concentration is network-driven and activity-dependent, as it is blocked by the non-NMDA glutamate receptor antagonist DNQX. We also show that expression of the outward K-Cl cotransporter, KCC2, prevents the development of hyperexcitability, as a reduction of KCC2 expression by half results in increased susceptibility to seizure under control and 4-AP conditions. PMID- 18394866 TI - PDE4B polymorphisms and decreased PDE4B expression are associated with schizophrenia. AB - Schizophrenia has a complex genetic underpinning and variations in a number of candidate genes have been identified that confer risk of developing the disorder. We report in the present studies that several single nucleotide polymorphisms (SNPs) and a two-SNP haplotype in PDE4B are associated with an increased incidence of schizophrenia in two large populations of Caucasian and African American patients. The SNPs in PDE4B associated with schizophrenia occur in intronic sequences in the vicinity of a critical splice junction that gives rise to the expression of PDE4B isoforms with distinct regulation and function. We also observed specific decreases in phosphodiesterase 4B (PDE4B) isoforms in brain tissue obtained postmortem from patients diagnosed with schizophrenia and bipolar disorder. PDE4B metabolically inactivates the second messenger cAMP to regulate intracellular signaling in neurons throughout the brain. Thus, the present observations suggest that dysregulation of intracellular signaling mediated by PDE4B is a significant factor in the cause and expression, respectively, of schizophrenia and bipolar disorder and that targeting PDE4B regulated signaling pathways may yield new therapies to treat the totality of these disorders. PMID- 18394865 TI - Long-term decrease in calbindin-D28K expression in the hippocampus of epileptic rats following pilocarpine-induced status epilepticus. AB - Acquired epilepsy (AE) is characterized by spontaneous recurrent seizures and long-term changes that occur in surviving neurons following an injury such as status epilepticus (SE). Long-lasting alterations in hippocampal Ca(2+) homeostasis have been observed in both in vivo and in vitro models of AE. One major regulator of Ca(2+) homeostasis is the neuronal calcium binding protein, calbindin-D28k that serves to buffer and transport Ca(2+) ions. This study evaluated the expression of hippocampal calbindin levels in the rat pilocarpine model of AE. Calbindin protein expression was reduced over 50% in the hippocampus in epileptic animals. This decrease was observed in the pyramidal layer of CA1, stratum lucidum of CA3, hilus, and stratum granulosum and stratum moleculare of the dentate gyrus when corrected for cell loss. Furthermore, calbindin levels in individual neurons were also significantly reduced. In addition, the expression of calbindin mRNA was decreased in epileptic animals. Time course studies demonstrated that decreased calbindin expression was initially present 1 month following pilocarpine-induced SE and lasted for up to 2 years after the initial episode of SE. The results indicate that calbindin is essentially permanently decreased in the hippocampus in AE. This decrease in hippocampal calbindin may be a major contributing factor underlying some of the plasticity changes that occur in epileptogenesis and contribute to the alterations in Ca(2+) homeostasis associated with AE. PMID- 18394867 TI - Striatal dopaminergic terminals in type 1 and type 2 alcoholics measured with [3H]dihydrotetrabenazine and human whole hemisphere autoradiography. AB - A number of studies have pointed to the importance of dopamine system in the context of alcoholism. Previous studies have shown lower dopamine transporter levels on late-onset Cloninger type 1 alcoholics. However, whether this lower level is due to a lower level of dopamine transporter protein or a lower level of dopaminergic nerve terminals remains unclear. The aim of this study was to compare putative alterations of dopaminergic terminals in caudate, putamen and nucleus accumbens of type 1 and type 2 alcoholics and healthy controls by using [(3)H]dihydrotetrabenazine as a radioligand in postmortem human whole hemisphere autoradiography. We compared the present results with the findings of our earlier studies on the dopamine transporter in these same subjects, demonstrating that alcoholics do not differ significantly from controls in striatal [(3)H]dihydrotetrabenazine binding. Although type 1 alcoholics have been reported to have up to 36% lower striatal dopamine transporter levels than controls, the results suggest that the density of their dopaminergic nerve terminals is not altered. PMID- 18394869 TI - A new principal component analysis-based approach for testing "similarity" of drug dissolution profiles. AB - A new approach for testing batch "similarity" through comparison of drug dissolution profiles, based on principal component analysis with the establishment of a confidence region (PCA-CR), is presented. The dissolution curves corresponding to three brands each of Furosemide and Acetaminophen tablets, taken as model drugs, were prepared by dissolution measurements at multiple pre-specified time points. Reference and test data were simultaneously subjected to PCA and pairwise comparisons between the dissolution characteristics of lots of the same and different brands were carried out. The comparisons involved plotting the weighed scores of the first two principal components of reference and test lots, while decision about "similarity" was made by checking for inclusion of more than 80% of the tablets of the test lot in the 95% confidence ellipse of the reference samples. Two published datasets were also analyzed in the same fashion and all the results were compared with information provided by the difference (f1) and similarity (f2) factor tests. Unlike the f2 criterion, the proposed method reflects variability within the individual dissolution curves, being also highly sensitive to profile (shape and size) variations. Comparison between the area enclosed by the confidence ellipses of the weighed scores plot and the region obtained from the bootstrap-calculated acceptable values of the corresponding f2 tests suggested that PCA-CR represents, in general, a more discriminating standard. PMID- 18394868 TI - Catch and release cell sorting: electrochemical desorption of T-cells from antibody-modified microelectrodes. AB - The development of integrated microsystems capable of interrogation, characterization and sorting of mammalian cells is highly significant for further advancement of point-of-care diagnostics and drug discovery fields. The present study sought to design a novel strategy for releasing antibody-bound cells through electrochemical disruption of the underlying antibody (Ab) layer. A microsystem for selective capture and release of cells consisted of an array of individually addressable gold microelectrodes fabricated on a glass substrate. Poly(ethylene glycol) (PEG) hydrogel photolithography was employed to make the glass regions non-fouling, thus, ensuring selective localization of proteins and cells on the microelectrodes. The gold surfaces were decorated with anti-CD4 Ab molecules using standard alkanethiol self-assembly and carbodiimide coupling approaches. The Ab-functionalized electrodes selectively captured model T lymphocytes (Molt-3 cells) expressing CD4 antigen while minimal cell adhesion was observed on PEG hydrogel-modified glass substrates. Importantly, application of a reductive potential (-1.2V vs. Ag/AgCl reference electrode) resulted in release of surface-bound T-cells from the electrode surface. Cyclic voltammetry and fluorescence microscopy were employed to verify that the detachment of captured T cells was indeed due to the electrochemical disruption of the underlying alkanethiol-Ab layer. In the future, the cell sorting approach described here may be combined with microfluidic delivery to enable Ab-mediated capture of T lymphocytes or other cell types followed by release of select cells for downstream gene expression studies or re-cultivation. PMID- 18394870 TI - Salinity tolerance in Hyalophysa chattoni (Ciliophora, Apostomatida), a symbiont of the estuarine grass shrimp Palaemonetes pugio. AB - The apostome ciliate Hyalophysa chattoni, a symbiont of the estuarine grass shrimp Palaemonetes pugio, was tested for its growth and reproductive ability in a wide range of salinities from 0.1 to 55 ppt. Shrimp, with their attached ciliates, were slowly acclimated to different salinities in order to assess protozoan cell size and division. The trophont and tomont stages of the ciliate life cycle were analyzed. In both stages, cell size increased with salinity from 0.1 to 20 ppt. Cell size leveled in the 20-35 ppt range, and decreased at higher salinities. The number of daughter cells produced per tomont cyst correlated with increased cell size, and also correlated with increased salinity. Additionally, increased salinity correlated with an increase in the percentage of cells able to divide and excyst as tomite stages. These results indicate that H. chattoni is able to grow and divide more effectively at salinities closer to seawater than in the estuarine environment from which they were collected. Though able to survive salinities from 0.1 to 55 ppt, the species is better adapted for an existence in the higher salt concentrations. PMID- 18394871 TI - Discovery and integration of univariate patterns from daily individual organ failure scores for intensive care mortality prediction. AB - OBJECTIVES: The current established mortality predictive models in the intensive care rely only on patient information gathered within the first 24 hours of admission. Recent research demonstrated the added prognostic value residing in the sequential organ-failure assessment (SOFA) score which quantifies on each day the cumulative patient organ derangement. The objective of this paper is to develop and study predictive models that also incorporate univariate patterns of the six individual organ systems underlining the SOFA score. A model for a given day d predicts the probability of in-hospital mortality. MATERIALS AND METHODS: We use the logistic framework to combine a summary statistic of the historic SOFA information for a patient together with selected dummy variables indicating the occurrence of univariate frequent temporal patterns of individual organ system functioning. We demonstrate the application of our method to a large real-life data set from an intensive care unit (ICU) in a teaching hospital. Model performance is tested in terms of the AUC and the Brier score. RESULTS: An algorithm for categorization, discovery, and selection of univariate patterns of individual organ scores and the induction of predictive models. The case-study resulted in six daily models corresponding to days 2-7. Their AUC ranged between 0.715 and 0.794 and the Brier scores between 0.161 and 0.216. Models using only admission data but recalibrated for days 2-7 generated AUC ranging between 0.643 and 0.761 and Brier scores ranged between 0.175 and 0.230. CONCLUSIONS: The results show that temporal organ-failure episodes improve predictions' quality in terms of both discrimination and calibration. In addition, they enhance the interpretability of models. Our approach should be applicable to many other medical domains where severity scores and sub-scores are collected. PMID- 18394872 TI - Sexual intercourse and respiratory failure. AB - Sexual activity is an important component of quality of life in patients suffering from chronic illnesses. To our knowledge, the effects of sexual activity on gas exchange in patients with respiratory failure have not been yet studied. To such an extent, we evaluated the oxygen saturation (SaO2), by a pulse oxymeter, during three different sexual performances in a 63-yr-old patient affected by chronic obstructive pulmonary disease (COPD) on long-term oxygen therapy (LTOT). The sexual performances were divided in four periods: basal, sex, 10 min after sex and relax. In each performance during sex, we observed a significant increase of either heart rate (HR) or SaO2, with the highest value of the latter achieved within the 10 min of the post-sex period. SaO2 returned to basal value (pre-sex) by the end of the relax period. We conclude that the observed improvement of SaO2 during sexual activity might be due to a better ventilation/perfusion ratio (V/Q) obtained for either an increase of ventilation (hyperventilation) and perfusion (tachycardia), without significant muscle expenditure. PMID- 18394873 TI - Reduced number and activity of circulating endothelial progenitor cells in patients with idiopathic pulmonary arterial hypertension. AB - BACKGROUND: Endothelial dysfunction plays a central and critical role in the initiation and development of idiopathic pulmonary arterial hypertension (IPAH), and a variety of evidence suggests that endothelial progenitor cells (EPCs) constitute one aspect of endothelium repair. In addition, transplantation of EPCs could attenuate pulmonary hypertension induced by monocrotaline in rats. However, it has not been examined and reported whether circulating EPCs from patients with IPAH are damaged. METHODS: EPCs were isolated and cultured from patients with IPAH (n=20) and matched healthy volunteers (n=20). Circulating EPC numbers (enumerated as AC133+KDR+ cells) as well as migratory and adhesive activity were assessed. Blood levels of vascular endothelial growth factor (VEGF), homocysteine (Hcy), B-type natriuretic peptide (BNP), von Willebrand Factor (vWF) and interleukin-6 (IL-6) were also measured. RESULTS: A significant decrease was observed in circulating EPC (AC133+KDR+ cells, 86.6+/-20.7cells/ml blood vs. 119.6+/-25.4cells/ml blood, P<0.001) numbers and the cell numbers expanded in vitro (47.2+/-14.5 vs. 70.7+/-15.2EPCs/x200 field; P<0.001) in patients with IPAH. EPCs from patients with IPAH were significantly impaired in their migratory capacity and ability to adhere to fibronectin. Blood levels of VEGF, Hcy, BNP, vWF and IL-6 were elevated in patients with IPAH. EPC numbers and activity were inversely related to Hcy, IL-6, BNP and vWF. CONCLUSIONS: Our observations indicated that EPC numbers and functional capacity were impaired in patients with IPAH, which might not only give potential insight into the pathophysiological mechanisms but also might be useful for identifying suitable therapeutic targets in these patients. PMID- 18394874 TI - Benzodiazepine and opioid sedation attenuate the sympathetic response to fiberoptic bronchoscopy. Prophylactic labetalol gave no additional benefit. Results of a randomized double-blind placebo-controlled study. AB - BACKGROUND: Hypertension and tachycardia are common during fiber-optic bronchoscopy (FOB), and this may lead to cardiac ischemia. The prophylactic addition of a beta-adrenergic anatagonist might mask this response and prevent the deleterious cardiovascular effects of FOB. METHODS: We performed a randomized double-blind placebo-controlled trial of labetalol 10mg iv given with midazolam alfentanil sedation. We monitored heart rate (HR) and systolic/diastolic blood pressure (SBP/DBP) throughout the bronchoscopy and calculated the rate-pressure product (RPP=(HRxSBP)/100). One-hundred twenty patients were enrolled. RESULTS: In the placebo group, there was no rise in HR, SBP, DBP or RPP, and there was no difference between the placebo and labetalol groups. Adverse events during bronchoscopy were similar in both groups. In a subgroup of patients undergoing interventional bronchoscopy, there was a trend towards lower SBP (p=0.06). CONCLUSIONS: Patients undergoing FOB under adequate midazolam-alfentanil sedation do not develop excessive sympathetic drive that may lead to cardiac stress. The addition of labetalol did not confer additional benefit or risk to the patients. (ClinicalTrials.gov number, NCT00394537). PMID- 18394875 TI - Comparison of salmeterol/fluticasone propionate (FP) combination with FP+sustained release theophylline in moderate asthma patients. AB - OBJECTIVES: To compare the efficacy and safety of the salmeterol/fluticasone propionate combination product with concurrent sustained release theophylline plus fluticasone propionate in adult Japanese patients with persistent asthma. DESIGN: Multicentre, randomised, double-blind, double-dummy, parallel-group study. PATIENTS AND INTERVENTIONS: Three hundred and eighty-three asthmatic patients receiving sustained release theophylline 200-400mg/day entered the study and were randomised to receive either salmeterol/fluticasone propionate combination (SFC) 50microg/250microg+1 placebo tablet, fluticasone propionate 250microg+1 sustained release theophylline 200mg (SR-T+FP), twice daily for 8 weeks. RESULTS: The adjusted mean change morning peak expiratory flow (PEF) over 8 weeks was 29.8L/min in the SFC group and 16.3L/min in the SR-T+FP group, with a treatment difference of 13.4L/min (p=0.0004). SFC improved evening PEF, FEV1, V50 and V25 at the completion of treatment to a greater extent than SR-T+FP (all p<0.05). A higher percentage of patients on SFC were symptom free (p=0.0286) and rescue free (ns) than those on SR-T+FP. There was not a statistically significant difference between treatments in symptom scores. Both treatments were well tolerated. CONCLUSIONS: The finding that SFC was associated with greater improvements in lung function than SR-T+FP, a commonly employed treatment for asthmatic patients in Japan, suggests that SFC should be the preferred therapeutic option in these patients. PMID- 18394877 TI - Cell regulation. PMID- 18394876 TI - FoxO transcription factors in the maintenance of cellular homeostasis during aging. AB - The FoxO family of Forkhead transcription factors functions at the interface of tumor suppression, energy metabolism, and organismal longevity. FoxO factors are key downstream targets of insulin, growth factor, nutrient, and oxidative stress stimuli that coordinate a wide range of cellular outputs. FoxO-dependent cellular responses include gluconeogenesis, neuropeptide secretion, atrophy, autophagy, apoptosis, cell cycle arrest, and stress resistance. This review will discuss the roles of the mammalian FoxO family in a variety of cell types, from stem cells to mature cells, in the context of the whole organism. Given the overwhelming evidence that the FoxO factors promote longevity in invertebrates, this review will also discuss the potential role of the FoxO factors in the aging of mammalian organisms. PMID- 18394878 TI - Metabolic engineering of plant volatiles. AB - Metabolic engineering of the volatile spectrum offers enormous potential for plant improvement because of the great contribution of volatile secondary metabolites to reproduction, defense and food quality. Recent advances in the identification of the genes and enzymes responsible for the biosynthesis of volatile compounds have made this metabolic engineering highly feasible. Notable successes have been reported in enhancing plant defenses and improving scent and aroma quality of flowers and fruits. These studies have also revealed challenges and limitations which will be likely surmounted as our understanding of plant volatile network improves. PMID- 18394879 TI - Tamoxifen exposure in relation to gastric adenocarcinoma development. AB - Epidemiological research has indicated that the anti-oestrogen tamoxifen, used in breast cancer therapy, may increase the risk of gastric adenocarcinoma of the intestinal but not of the diffuse type. To test this hypothesis, and evaluate possible involvement of oestrogen receptors (ERs), we conducted a study amongst tamoxifen users and non-users. The study participants comprised women in the county of Stockholm who in the Swedish Cancer Register were first recorded with breast cancer and subsequently gastric cancer during the period January 1958 August 2005. Medical records were scrutinised to verify the diagnoses and classify into use or non-use of tamoxifen. Tumour material was reviewed histologically to verify gastric adenocarcinoma diagnosis and classify these cancers into intestinal or diffuse type. Intestinal adenocarcinomas were analysed immunohistochemically for the presence of ER alpha, beta and beta cx. Amongst 68 women with verified gastric adenocarcinoma, 30 had been treated with tamoxifen and 38 not. The intestinal type of gastric adenocarcinoma was not more frequent amongst tamoxifen users (27%) than amongst non-users (34%) (p=0.601). There were no material differences between the tamoxifen groups regarding distribution of any of the three ERs of the intestinal adenocarcinoma specimens. Tamoxifen users had a shorter latency between breast cancer and gastric adenocarcinoma (4 versus 13 years) which was similar in the intestinal and diffuse types. This study does not support the hypothesis that tamoxifen increases the isolated risk of the intestinal type, but it indicates that tamoxifen use might accelerate the tumour progression or increase the overall risk of gastric adenocarcinoma. PMID- 18394880 TI - Predicting survival in patients with advanced disease. AB - Prognostication is an important clinical skill for all clinicians, particularly those clinicians working with patients with advanced cancer. However, doctors can be hesitant about prognosticating without a fundamental understanding of how to formulate a prognosis more accurately and how to communicate the information with honesty and compassion. Irrespective of the underlying type of malignancy, most patients with advanced cancer experience a prolonged period of gradual decline (months/years) before a short phase of accelerated decline in the last month or two. The main indicators of this final phase are poor performance status, weight loss, symptoms such as anorexia, breathlessness or confusion and abnormalities on laboratory parameters (e.g. high white cell count, lymphopaenia, hyopalbuminaemia, elevated lactate dehydrogenase or C-reactive protein). The clinical estimate of survival remains a powerful independent prognostic indicator, often enhanced by experience, but research has only begun to understand the different biases affecting clinicians' estimates. More recent research has shown probabilistic predictions to be more accurate than temporal predictions. Simple, reliable and valid prognostic tools have been developed in recent years that can be used readily at the bedside of terminally ill cancer patients. The greatest accuracy occurs with the use of a combination of subjective prognostic judgements and objective validated tools. Communicating survival predictions is an important part of cancer care and guidelines exist for improving delivery of such information. Important cultural differences may influence communication strategies and should be recognised in clinical encounters. More well-designed studies of prognosis and its impact on decision making are needed. The benefits and limitations of prognostication should be considered in many clinical decisions. PMID- 18394881 TI - A comparison of CDKN2A mutation detection within the Melanoma Genetics Consortium (GenoMEL). AB - CDKN2A is the major melanoma susceptibility gene so far identified, but only 40% of three or more case families have identified mutations. A comparison of mutation detection rates was carried out by "blind" exchange of samples across GenoMEL, the Melanoma Genetics Consortium, to establish the false negative detection rates. Denaturing high performance liquid chromatography (DHPLC) screening results from 451 samples were compared to screening data from nine research groups in which the initial mutation screen had been done predominantly by sequencing. Three samples with mutations identified at the local centres were not detected by the DHPLC screen. No additional mutations were detected by DHPLC. Mutation detection across groups within GenoMEL is carried out to a consistently high standard. The relatively low rate of CDKN2A mutation detection is not due to failure to detect mutations and implies the existence of other high penetrance melanoma susceptibility genes. PMID- 18394882 TI - Efficient lipid production with Trichosporon fermentans and its use for biodiesel preparation. AB - Effects of medium components and culture conditions on biomass and lipid production of Trichosporon fermentans were studied. The optimal nitrogen source, carbon source and C/N molar ratio were peptone, glucose and 163, respectively. The favorable initial pH of the medium and temperature were 6.5 and 25 degrees C. Under the optimized conditions, a biomass of 28.1 g/l and a lipid content of 62.4% could be achieved after culture for 7 days, which were much higher than the original values (19.4 g/l and 50.8%) and the results reported by other groups. T. fermentans could grow well in pretreated waste molasses and a lipid yield of 12.8 g/l could be achieved with waste molasses of 15% total sugar concentration (w/v) at pH 6.0, representing the best result with oleaginous microorganisms on agro industrial residues. Addition of various sugars to the pretreated molasses could efficiently enhance the accumulation of lipid and the lipid content reached as high as above 50%. Similar to vegetable oils, the lipid mainly contains palmitic acid, stearic acid, oleic acid and linoleic acid and the unsaturated fatty acids amount to about 64% of the total fatty acids. The microbial oil with an acid value of 5.6 mg KOH/g was transesterified to biodiesel by base catalysis after removal of free fatty acids and a high methyl ester yield of 92% was obtained. PMID- 18394883 TI - Three-step biological process for the treatment of the liquid fraction of cattle manure. AB - The liquid fraction of cattle manure was subjected to a biological treatment combining anoxic-anaerobic and oxic processes in order to stabilize the organic matter and reduce nitrogen and phosphorus so as to avoid problems of pollution when applying it to the land. The anoxic process was carried out at 30 degrees C in a CSTR reactor, the anaerobic process in a UASB reactor at 37 degrees C and the oxic treatment in another CSTR at 20 degrees C. The following results were obtained when working under optimum conditions (removal efficiencies in brackets): COD was reduced from 42 to 3.8 g/L (>90%); total solids from 41 to 14 (67%); total volatile solids from 22 to 7.0 (68%); total Kjeldahl nitrogen from 2.2 to 0.1 g/L (95%); NH4(-)-N from 1.10 to 0.02 g/L (98%) and Total-Phosphorus from 0.696 to 0.058 g/L (92%). Nitrates, undetected in the liquid fraction of cattle manure, were present in the final effluent as a result of nitrification. To reduce the amount of nitrates, different recirculation rates were tested. The minimum nitrate concentration achieved was 127 mg/L using a recirculation ratio of 4. PMID- 18394884 TI - Small molecules with structural similarities to siderophores as novel antimicrobials against Mycobacterium tuberculosis and Yersinia pestis. AB - Drugs inhibiting the iron scarcity-induced, siderophore-mediated iron-scavenging systems of Mycobacterium tuberculosis (Mtb) and Yersinia pestis (Yp) may provide new therapeutic lines of defense. Compounds with structural similarities to siderophores were synthesized and evaluated as antimicrobials against Mtb and Yp under iron-limiting conditions, which mimic the iron scarcity these pathogens encounter and must adapt to in the host, and under standard iron-rich conditions for comparison. New antimicrobials were identified, some of which warrant exploration as initial leads against potentially novel targets and small-molecule tools to assist in the elucidation of targets specific to iron-scarcity adapted Mtb and Yp. PMID- 18394885 TI - Microarray-based label-free detection of RNA using bispyrene-modified 2'-O-methyl oligoribonucleotide as capture and detection probe. AB - A novel oligonucleotide microarray that can detect RNAs without fluorescent labeling of sample RNAs was developed. As a capture and detection probe, bispyrene-modified 2'-O-methyl oligoribonucleotide (OMUpy2), whose fluorescence was dramatically increased when hybridized with its complementary RNA, was adopted. Fluorescence of the OMUpy2 tethered on the glass surface was enhanced as much as 22-fold by the addition of complementary oligoribonucleotide. PMID- 18394886 TI - 4'-Methyl-4,5'-bithiazole-based correctors of defective delta F508-CFTR cellular processing. AB - The synthesis and Delta F508-CFTR corrector activity of a 148-member methylbithiazole-based library are reported. Synthetic routes were devised and optimized to generate methylbithiazole analogs in four steps. Corrector potency and efficacy were assayed using epithelial cells expressing human Delta F508 CFTR. These structure-activity data establish that the bithiazole substructure plays a critical function; eight novel methylbithiazole correctors were identified with low micromolar potencies. PMID- 18394887 TI - 5-Sulfonyl-benzimidazoles as selective CB2 agonists. AB - A novel series of benzimidazole CB2-receptor agonists was synthesized and the structure-activity relationship explored. The results showed agonistic activities with an EC(50) up to 0.5 nM and excellent selectivity (>4000-fold) over the CB1 receptor. The size of the substituent on the 2-position determined the level of agonism, ranging from inverse agonism to partial agonism to full agonism, which was more pronounced for the rat CB2 receptor. A wide variation of sulfonyl substituents at the benzimidazole 5-position was tolerated, which was used to optimize the drug-like properties. This resulted into lead compound 14j that can be used to investigate the potential of a selective, peripherically acting CB2 agonist. The in vitro profile of key compounds is displayed using pie bar charts (VlaaiVis). PMID- 18394888 TI - A novel mutation in the dynamin 2 gene in a Charcot-Marie-Tooth type 2 patient: clinical and pathological findings. AB - Mutations in dynamin 2 (DNM2) have been associated with autosomal dominant centronuclear myopathy, dominant intermediate Charcot-Marie-Tooth (CMT) type B and CMT2. Here, we report a novel DNM2 mutation in the Pleckstrin homology domain of DNM2 (p.K559del) in a patient with an axonal length-dependent sensorimotor polyneuropathy predominantly affecting the lower limbs. Neuropathy is associated with congenital cataracts, ophthalmoparesis, ptosis and neutropenia. There was no evidence of a skeletal myopathy on EMG or muscle biopsy. We suggest that this constellation of clinical features can help the diagnosis and selection of patients for direct DNM2 genetic analysis. PMID- 18394889 TI - Evidence for degeneration of the Y chromosome in the dioecious plant Silene latifolia. AB - The human Y--probably because of its nonrecombining nature--has lost 97% of its genes since X and Y chromosomes started to diverge [1, 2]. There are clear signs of degeneration in the Drosophila miranda neoY chromosome (an autosome fused to the Y chromosome), with neoY genes showing faster protein evolution [3-6], accumulation of unpreferred codons [6], more insertions of transposable elements [5, 7], and lower levels of expression [8] than neoX genes. In the many other taxa with sex chromosomes, Y degeneration has hardly been studied. In plants, many genes are expressed in pollen [9], and strong pollen selection may oppose the degeneration of plant Y chromosomes [10]. Silene latifolia is a dioecious plant with young heteromorphic sex chromosomes [11, 12]. Here we test whether the S. latifolia Y chromosome is undergoing genetic degeneration by analyzing seven sex-linked genes. S. latifolia Y-linked genes tend to evolve faster at the protein level than their X-linked homologs, and they have lower expression levels. Several Y gene introns have increased in length, with evidence for transposable-element accumulation. We detect signs of degeneration in most of the Y-linked gene sequences analyzed, similar to those of animal Y-linked and neo-Y chromosome genes. PMID- 18394890 TI - JNK signaling controls border cell cluster integrity and collective cell migration. AB - Collective cell movement is a mechanism for invasion identified in many developmental events. Examples include the movement of lateral-line neurons in Zebrafish, cells in the inner blastocyst, and metastasis of epithelial tumors [1]. One key model to study collective migration is the movement of border cell clusters in Drosophila. Drosophila egg chambers contain 15 nurse cells and a single oocyte surrounded by somatic follicle cells. At their anterior end, polar cells recruit several neighboring follicle cells to form the border cell cluster [2]. By stage 9, and over 6 hr, border cells migrate as a cohort between nurse cells toward the oocyte. The specification and directionality of border cell movement are regulated by hormonal and signaling mechanisms [3]. However, how border cells are held together while they migrate is not known. Here, we show that a negative-feedback loop controlling JNK activity regulates border cell cluster integrity. JNK signaling modulates contacts between border cells and between border cells and substratum to sustain collective migration by regulating several effectors including the polarity factor Bazooka and the cytoskeletal adaptor D-Paxillin. We anticipate a role for the JNK pathway in controlling collective cell movements in other morphogenetic and clinical models. PMID- 18394891 TI - Regulation of cell adhesion and collective cell migration by hindsight and its human homolog RREB1. AB - Cell movements represent a major driving force in embryonic development, tissue repair, and tumor metastasis [1]. The migration of single cells has been well studied, predominantly in cell culture [2, 3]; however, in vivo, a greater variety of modes of cell movement occur, including the movements of cells in clusters, strands, sheets, and tubes, also known as collective cell migrations [4, 5]. In spite of the relevance of these types of movements in both normal and pathological conditions, the molecular mechanisms that control them remain predominantly unknown. Epithelial follicle cells of the Drosophila ovary undergo several dynamic morphological changes, providing a genetically tractable model [6]. We found that anterior follicle cells, including border cells, mutant for the gene hindsight (hnt) accumulated excess cell-cell adhesion molecules and failed to undergo their normal collective movements. In addition, HNT affected border cell cluster cohesion and motility via effects on the JNK and STAT pathways, respectively. Interestingly, reduction of expression of the mammalian homolog of HNT, RREB1, by siRNA inhibited collective cell migration in a scratch wound healing assay of MCF10A mammary epithelial cells, suppressed surface activity, retarded cell spreading after plating, and led to the formation of immobile, tightly adherent cell colonies. We propose that HNT and RREB1 are essential to reduce cell-cell adhesion when epithelial cells within an interconnected group undergo dynamic changes in cell shape. PMID- 18394892 TI - ARF GEF-dependent transcytosis and polar delivery of PIN auxin carriers in Arabidopsis. AB - Cell polarity manifested by the polar cargo delivery to different plasma-membrane domains is a fundamental feature of multicellular organisms. Pathways for polar delivery have been identified in animals; prominent among them is transcytosis, which involves cargo movement between different sides of the cell [1]. PIN transporters are prominent polar cargoes in plants, whose polar subcellular localization determines the directional flow of the signaling molecule auxin [2, 3]. In this study, we address the cellular mechanisms of PIN polar targeting and dynamic polarity changes. We show that apical and basal PIN targeting pathways are interconnected but molecularly distinct by means of ARF GEF vesicle trafficking regulators. Pharmacological or genetic interference with the Arabidopsis ARF GEF GNOM leads specifically to apicalization of basal cargoes such as PIN1. We visualize the translocation of PIN proteins between the opposite sides of polarized cells in vivo and show that this PIN transcytosis occurs by endocytic recycling and alternative recruitment of the same cargo molecules by apical and basal targeting machineries. Our data suggest that an ARF GEF dependent transcytosis-like mechanism is operational in plants and provides a plausible mechanism to trigger changes in PIN polarity and hence auxin fluxes during embryogenesis and organogenesis. PMID- 18394893 TI - Wind selection and drift compensation optimize migratory pathways in a high flying moth. AB - Numerous insect species undertake regular seasonal migrations in order to exploit temporary breeding habitats [1]. These migrations are often achieved by high altitude windborne movement at night [2-6], facilitating rapid long-distance transport, but seemingly at the cost of frequent displacement in highly disadvantageous directions (the so-called "pied piper" phenomenon [7]). This has lead to uncertainty about the mechanisms migrant insects use to control their migratory directions [8, 9]. Here we show that, far from being at the mercy of the wind, nocturnal moths have unexpectedly complex behavioral mechanisms that guide their migratory flight paths in seasonally-favorable directions. Using entomological radar, we demonstrate that free-flying individuals of the migratory noctuid moth Autographa gamma actively select fast, high-altitude airstreams moving in a direction that is highly beneficial for their autumn migration. They also exhibit common orientation close to the downwind direction, thus maximizing the rectilinear distance traveled. Most unexpectedly, we find that when winds are not closely aligned with the moth's preferred heading (toward the SSW), they compensate for cross-wind drift, thus increasing the probability of reaching their overwintering range. We conclude that nocturnally migrating moths use a compass and an inherited preferred direction to optimize their migratory track. PMID- 18394894 TI - Cell-polarity dynamics controls the mechanism of lumen formation in epithelial morphogenesis. AB - Many organs consist of tubes of epithelial cells enclosing a central lumen. How the space of this lumen is generated is a key question in morphogenesis. Two predominant mechanisms of de novo lumen formation have been observed: hollowing and cavitation. In hollowing, the lumen is formed by exocytosis and membrane separation, whereas, in cavitation, the lumen is generated by apoptosis of cells in the middle of the structure [1, 2]. Using MDCK cells in three-dimensional cultures, we found an inverse correlation between polarization efficiency and apoptosis. When cells were grown in collagen, where cells polarized slowly, apoptosis was needed for lumen formation. However, in the presence of Matrigel, which allowed rapid polarization, lumens formed without apoptosis. If polarization in Matrigel was perturbed by blocking formation of the apical surface by RNAi of Cdc42, lumens formed by apoptosis. In a complementary approach, we plated cells at high density so that aggregates formed with little polarity. These aggregates required apoptosis to form lumens, whereas cells plated at low density formed cysts with rapidly polarizing cells and did not need apoptosis to form lumens. The mechanism of lumen formation in the 3D-MDCK model can shift between hollowing and cavitation, depending on cell polarization. PMID- 18394895 TI - Temporal reciprocity of miRNAs and their targets during the maternal-to-zygotic transition in Drosophila. AB - During oogenesis, female animals load their eggs with messenger RNAs (mRNAs) that will be translated to produce new proteins in the developing embryo. Some of these maternally provided mRNAs are stable and continue to contribute to development long after the onset of transcription of the embryonic (zygotic) genome. However, a subset of maternal mRNAs are degraded during the transition from purely maternal to mixed maternal-zygotic gene expression. In Drosophila, two independent RNA degradation pathways are used to promote turnover of maternal transcripts during the maternal-to-zygotic transition [1]. The first is driven by maternally encoded factors, including SMAUG [2], whereas the second is activated about 2 hr after fertilization, coinciding with the onset of zygotic transcription. Here, we report that a cluster of zygotically expressed microRNAs (miRNAs) targets maternal mRNAs for turnover, as part of the zygotic degradation pathway. miRNAs are small noncoding RNAs that silence gene expression by repressing translation of their target mRNAs and by promoting mRNA turnover. Intriguingly, use of miRNAs to promote mRNA turnover during the maternal-to zygotic transition appears to be a conserved phenomenon because a comparable role was reported for miR-430 in zebrafish [3]. The finding that unrelated miRNAs regulate the maternal to zygotic transition in different animals suggests convergent evolution. PMID- 18394896 TI - Flap endonuclease 1 contributes to telomere stability. AB - Telomere stability plays an important role in the preservation of genomic stability and is maintained through the coordinated actions of telomere-specific proteins and DNA repair and replication proteins [1, 2]. Flap endonuclease 1 (FEN1) is a protein that plays a role in lagging-strand DNA replication, base excision repair, homologous recombination, and reinitiation of stalled replication forks [3, 4]. Here, we demonstrate that FEN1 depletion leads to telomere dysfunction characterized by the presence of gammaH2AX and sister telomere loss. Expression of catalytically active telomerase, the reverse transcriptase that adds telomeric repeats to chromosome ends, was sufficient to rescue telomere dysfunction upon FEN1 depletion. Strikingly, FEN1 depletion exclusively abrogates telomeres replicated by lagging-strand DNA replication. Genetic rescue experiments utilizing FEN1 mutant proteins that retained the ability to localize to telomeric repeats revealed that FEN1's nuclease activity and ability to interact with the Werner protein (WRN) and telomere-binding protein (TRF2) were required for FEN1 activity at the telomere. Given FEN1's role in lagging-strand DNA replication and reinitiation of stalled replication forks, we propose that FEN1 contributes to telomere stability by ensuring efficient telomere replication. PMID- 18394897 TI - RNA interference guides histone modification during the S phase of chromosomal replication. AB - BACKGROUND: Heterochromatin is chromosomal material that remains condensed throughout the cell division cycle and silences genes nearby. It is found in almost all eukaryotes, and although discovered (in plants) almost 100 years ago, the mechanism by which heterochromatin is inherited has remained obscure. Heterochromatic silencing and histone H3 lysine-9 methylation (H3K9me2) depend, paradoxically, on heterochromatic transcription and RNA interference (RNAi). RESULTS: Here, we show that heterochromatin protein 1 in fission yeast (Swi6) is lost via phosphorylation of H3 serine 10 (H3S10) during mitosis, allowing heterochromatic transcripts to transiently accumulate in S phase. Rapid processing of these transcripts into small interfering RNA (siRNA) promotes restoration of H3K9me2 and Swi6 after replication when cohesin is recruited. We also show that RNAi in fission yeast is inhibited at high temperatures, providing a plausible mechanism for epigenetic phenomena that depend on replication and temperature, such as vernalization in plants and position effect variegation in animals. CONCLUSIONS: These results explain how "silent" heterochromatin can be transcribed and lead to a model for epigenetic inheritance during replication. PMID- 18394899 TI - Cellular functions of GEF-H1, a microtubule-regulated Rho-GEF: is altered GEF-H1 activity a crucial determinant of disease pathogenesis? AB - The Rho guanine nucleotide exchange factor GEF-H1 is uniquely regulated by microtubule binding and is crucial in coupling microtubule dynamics to Rho-GTPase activation in a variety of normal biological situations. Here, we review the roles of GEF-H1 in epithelial barrier permeability, cell motility and polarization, dendritic spine morphology, antigen presentation, leukemic cell differentiation, cell cycle regulation, and cancer. GEF-H1 might also contribute to pathophysiological signaling involved in leukemias, and in cancers associated with mutated p53 tumor suppressor gene, epithelial and endothelial cell dysfunction, infectious disease, and cardiac hypertrophy. We suggest that GEF-H1 could be a novel therapeutic target in multiple human diseases. PMID- 18394898 TI - Distinct innate immune responses to infection and wounding in the C. elegans epidermis. AB - BACKGROUND: In many animals, the epidermis is in permanent contact with the environment and represents a first line of defense against pathogens and injury. Infection of the nematode Caenorhabditis elegans by the natural fungal pathogen Drechmeria coniospora induces the expression in the epidermis of antimicrobial peptide (AMP) genes such as nlp-29. Here, we tested the hypothesis that injury might also alter AMP gene expression and sought to characterize the mechanisms that regulate the innate immune response. RESULTS: Injury induces a wound-healing response in C. elegans that includes induction of nlp-29 in the epidermis. We find that a conserved p38-MAP kinase cascade is required in the epidermis for the response to both infection and wounding. Through a forward genetic screen, we isolated mutants that failed to induce nlp-29 expression after D. coniospora infection. We identify a kinase, NIPI-3, related to human Tribbles homolog 1, that is likely to act upstream of the MAPKK SEK-1. We find NIPI-3 is required only for nlp-29 induction after infection and not after wounding. CONCLUSIONS: Our results show that the C. elegans epidermis actively responds to wounding and infection via distinct pathways that converge on a conserved signaling cassette that controls the expression of the AMP gene nlp-29. A comparison between these results and MAP kinase signaling in yeast gives insights into the possible origin and evolution of innate immunity. PMID- 18394900 TI - Intermittent versus continuous renal replacement therapy: a matter of controversy. AB - BACKGROUND: Acute Renal Failure (ARF) requiring some form of replacement therapy is a frequent complication in the critically ill patient. Despite potential therapeutic advantages the expectation of an improvement in patient outcomes using Continuous Renal Replacement Therapy (CRRT) compared to conventional Intermittent Haemodialysis (IHD) remains controversial. AIMS AND METHOD: This article will review the literature on the issues surrounding the use of IHD versus CRRT in the management of the critically ill patient. Articles were selected according to level of evidence with priority given to meta-analyses and randomised controlled trials. DISCUSSION: Several operational features of CRRT allow this technique to be tolerated more easily in critical illness than IHD. The gradual removal of fluid reduces the incidence of hypotension and the risk of volume overload. Decreased variability in the concentration of solutes enables greater azotemia control. However, CRRT is required to operate uninterrupted to achieve a treatment dose that is equivalent to a conventional IHD treatment schedule. In the absence of definitive evidence to validate superior patient survival and return of renal function there is disagreement as to the most appropriate form of Renal Replacement Therapy (RRT) for the critically ill patient. The introduction of 'hybrid' therapies offers a further alternative treatment strategy, which combine favourable aspects of IHD and CRRT. CONCLUSION: The decision to use IHD or CRRT should be guided by the therapeutic needs of the patient rather than the operational differences between the two techniques. The resources and expertise available at the organisation are also important in determining the mode best able to manage the critically ill patient at any stage and may change according to the severity of illness. The emergence of hybrid therapies provides a compromise option which encompasses many of the features of both systems, but does not embrace all options of either approach. PMID- 18394902 TI - Structure and mode of action of clostridial glucosylating toxins: the ABCD model. AB - Toxins A and B, which are the major virulence factors of antibiotic-associated diarrhea and pseudomembranous colitis caused by Clostridium difficile, are the prototypes of the family of clostridial glucosylating toxins. The toxins inactivate Rho and Ras proteins by glucosylation. Recent findings on the autocatalytic processing of the toxins and analysis of the crystal structures of their domains have made a revision of the current model of their actions on the eukaryotic target cells necessary. PMID- 18394903 TI - Connecting loose ends: a unique linear plasmid or a new model system? PMID- 18394904 TI - Hypericin uptake: a prognostic marker for survival in high-grade glioma. AB - Currently adjuvant chemotherapy for glioblastoma patients can prolong survival time relative to patients who receive only surgery and radiotherapy. Despite these improvements and experimental and clinical efforts the prognosis for glioblastoma patients remains poor. At present, interest is focused on individual prognostic factors influencing patient responses to therapy. Photodynamic therapy may be a promising therapeutic option in the treatment of glioblastoma. In this investigation we examined whether uptake of hypericin (HY), a fluorescent photosensitization agent, by ex vivo glioblastoma cell lines correlates with prognosis of the individual from which the cell lines were derived. Twelve primary human glioma cell cultures were incubated with 20 micromol HY. Fluorescence intensity was measured using fluorescence microscopy. Three patients suffered from an anaplastic astrocytoma, WHO grade III, nine had a glioblastoma, WHO grade IV. In 6/12 patients complete tumour resection was possible. The mean survival time of the six patients in whom complete tumour resection was performed was 26 months, compared with 5 months for those who underwent incomplete resection. Eleven patients received radiation therapy. The five patients who received chemotherapy survived for a mean duration of 26 months, compared with the seven patients who survived for a mean duration of 5 months without chemotherapy. Statistical analysis using a parametric survival model based on the Weibull distribution showed that fluorescence intensity was the variable with the lowest p-value associated with survival (p=0.0225). An increase of 553 arbitrary units of fluorescence intensity is predicted to double survival time. Uptake of HY, a lipophilic molecule, is assumed to be related to low-density lipoprotein (LDL) uptake and metabolism. Cell proliferation is associated with a high turnover of cholesterol and membrane growth, which is related to cholesterol uptake by LDL. In summary, HY uptake by ex vivo glioblastoma cell cultures seems to be positively associated with survival of patients with malignant glioma. PMID- 18394905 TI - Primary spontaneous CSF rhinorrhea through the clivus: possible etiopathology. AB - Primary spontaneous cerebrospinal fluid (CSF) rhinorrhea is rare. Only two such cases with a defect in the clivus have been previously reported in the world literature. The purpose of this report is not to discuss the operative approach or results but the etiopathogenesis of the defect at this very rare site. One patient had a defect in the posterior wall of the sphenoid sinus, just caudal to the dorsum sellae. In the other, a psuedomeningocele (thickened arachnoid outpouching) was found protruding into the sphenoid sinus through a defect in the middle of the clivus. Both cases were successfully managed with transsphenoidal surgery. Two additional cases of primary spontaneous CSF rhinorrhea through a defect in clivus are reported. The possible mechanism of the fistulae are discussed. PMID- 18394906 TI - Synthesis of glutamic acid analogs as potent inhibitors of leukotriene A4 hydrolase. AB - Leukotriene B(4) (LTB(4)) is a potent pro-inflammatory mediator that has been implicated in the pathogenesis of multiple diseases, including psoriasis, inflammatory bowel disease, multiple sclerosis and asthma. As a method to decrease the level of LTB(4) and possibly identify novel treatments, inhibitors of the LTB(4) biosynthetic enzyme, leukotriene A(4) hydrolase (LTA(4)-h), have been explored. Here we describe the discovery of a potent inhibitor of LTA(4)-h, arylamide of glutamic acid 4f, starting from the corresponding glycinamide 2. Analogs of 4f are then described, focusing on compounds that are both active and stable in whole blood. This effort culminated in the identification of amino alcohol 12a and amino ester 6b which meet these criteria. PMID- 18394907 TI - Effects of modifications of the linker in a series of phenylpropanoic acid derivatives: Synthesis, evaluation as PPARalpha/gamma dual agonists, and X-ray crystallographic studies. AB - A new series of alpha-aryl or alpha-heteroarylphenyl propanoic acid derivatives was synthesized that incorporate acetylene-, ethylene-, propyl-, or nitrogen derived linkers as a replacement of the commonly used ether moiety that joins the central phenyl ring with the lipophilic tail. The effect of these modifications in the binding and activation of PPARalpha and PPARgamma was first evaluated in vitro. Compounds possessing suitable profiles were then evaluated in the ob/ob mouse model of type 2 diabetes. The propylene derivative 40 and the propyl derivative 53 demonstrated robust plasma glucose lowering activity in this model. Compound 53 was also evaluated in male Zucker diabetic fatty rats and was found to achieve normalization of glucose, triglycerides, and insulin levels. An X-ray crystal structure of the complex of 53 with the PPARgamma-ligand-binding domain was obtained and discussed in this report. PMID- 18394908 TI - Selective etching of dislocations in GaN and quantitative SEM analysis with shape reconstruction method. AB - Due to the differences in etch-pit morphologies, chemical etching offers a possibility to determine densities of dislocations in respect to their type. In the present paper we propose a method, which implements a simple shape-from shading procedure, i.e. with results derived from image brightness dependence on surface slope. It allows estimation of etch-pit depth distributions from scanning electron microscopy micrographs. This method is used to obtain depth distributions from GaN surface after etching in molten KOH-NaOH eutectic mixture. Depth distributions are used to estimate densities of etch-pits related to a given dislocation type. The distributions are compared with dislocation densities determined with transmission electron microscopy. PMID- 18394909 TI - Androgen receptor in the Mongolian gerbil ventral prostate: evaluation during different phases of postnatal development and following androgen blockage. AB - The normal growth, differentiation and maintenance of the morphofunctional integrity of the prostate gland are dependent on the interaction of constant levels of androgens with their receptors. The need to study the responses to hormones under several conditions and the effect of their blockage is due to the fact that the human prostate is the site of a great number of age-related diseases, and the ones with a major medical importance are prostate cancer (CaP) and benign prostatic hyperplasia (BPH), which can both be treated with androgen suppression. Seventy-five male gerbils were divided, randomly, into 3 groups of 25 animals each, where each group corresponded to one phase of postnatal development. In each phase, it was possible to morphologically and stereologically analyze the compartments of prostatic ventral lobe, as well as to immunohistochemically analyze the degree of expression of androgen receptors (ARs) after the androgen blockage therapies. In addition, it was possible to establish the hormonal dosage of serum testosterone levels given the comparative approach of the expression of androgen receptors. There is a pattern of AR distribution in the prostatic ventral lobe throughout postnatal development, in which the younger the animal is the higher, the interaction of circulating androgens that stimulate the AR expression in both the epithelial and stromal compartments. The androgen blockage therapies decreased AR expression in the prostatic compartments, but the androgen reposition after these blockages was not sufficient to recover the glandular structure or stimulate the AR expression up to normal physiological conditions. Both the regulation and distribution of androgen receptors along the gerbil prostatic tissues are complex mechanisms that are likely to be genetically regulated by androgens prenatally or by other factors that are still unknown. This rodent species seems to be a valuable model in the attempt to improve the understanding of the morphophysiological and pathological behavior of this important gland in humans throughout aging and to stimulate new therapeutic ideas to fight prostate cancer. PMID- 18394910 TI - Improved and semi-automated GMP-compliant radiosynthesis of [11C]docetaxel. AB - [(11)C]Docetaxel (Taxotere) has been synthesized via an improved synthesis route, which involves a more efficient intermediate removal of the excess 1,2,2,2 tetrachloroethyl chloroformate. Furthermore, the purification and formulation into a human applicable solution of [(11)C]docetaxel was developed. 1,2,2,2 Tetrachloroethyl chloroformate is used for the synthesis of the intermediate [(11)C]tert-butyl-1,2,2,2-tetrachloroethyl carbonate that in the final reaction step reacts with the precursor, the primary amine of docetaxel, yielding [(11)C]docetaxel. The purified and isolated product was obtained in 10+/-2% overall decay-corrected radiochemical yield. The total synthesis time was 67 min and the specific activity was 9-17 GBq/micromol (N=7). PMID- 18394911 TI - Measurement of (241)Am L X-ray emission probabilities. AB - Measurements of (241)Am L X-ray emission probabilities were conducted using both HPGe and Si(Li) detectors. The efficiency calibrations of these detectors were performed by means of a tunable, monochromatic photon beam and the determination of the thickness of absorbing materials inside the detector. These efficiency calibrations were obtained without any reference to radionuclide decay data, and with 0.8% relative standard uncertainty. The complex L X-ray region was processed using Voigt functions and by taking account of the detector response function established with the monochromatic beam. Twenty-two components of the L X-ray group were identified and quantified. The present results are compared with previously published data. PMID- 18394912 TI - Modified channel retractor for the sagittal split ramus osteotomy: a technical note. AB - The authors describe a modification of the commonly used inferior border channel retractor for the sagittal split osteotomy of the mandible. We modified this instrument by hollowing out the shank and working surface allowing improved access during the vertical cut through the inferior border of the mandible. Advantages of this modification over the original design include less dulling of the cutting instrument, less damage to the channel retractor, less debris in the surgical wound, and most importantly, less damage to the surrounding soft tissue. In our experience with this instrument, there have been no incidences of excessive blood loss requiring transfusion, avascular necrosis, bony nonunion or permanent nerve injury. We believe this modification improves manouverability during orthognathic surgery thereby decreasing morbidity and operating time, thereby making a successful outcome more likely. PMID- 18394913 TI - Thymoma appearing 10 years after an extended thymectomy for myasthenia gravis. AB - Occurrence of thymoma after an extended thymectomy through a full median sternotomy for nonthymomatous thymectomy has been very rarely reported. A 60-year old male patient who had myasthenia gravis (MG) for 11 years had an extended thymectomy operation with a pathology of thymic hyperplasia and developed a mass in the aortopulmonary window. We resected the mass via anterior left thoracotomy by dividing the ductus arteriosus and mobilizing the aorta. Cord vocal augmentation procedure was done due to the resection of the left recurrent laryngeal nerve. PMID- 18394914 TI - 'The way things are done around here'-Developing a culture of effectiveness: a pre-requisite to individual and team effectiveness in critical care. PMID- 18394916 TI - Glioblastoma in adults. AB - Glioblastoma (GBM) is the most malignant among astrocytic tumours and is associated with a poor prognosis. Age, performance status, mini-mental status examination score, methylation status of methylguanine methyltransferase promoter and extent of surgery constitute the main prognostic factors. Surgery aimed to complete resection should be the first therapeutic modality in the management of glioblastoma. However, complete resection is virtually impossible due to infiltrative nature of this disease and relapse is almost inevitable. Postoperative concomitant chemo-radiation is the standard treatment and consists of 60Gy of external-beam radiotherapy (to be delivered to a target volume including a 2-3cm ring of tissue surrounding the perimeter of the contrast enhancing lesion on pre-operative CT/MRI scans) plus temozolomide (TMZ) administered concomitantly (75mg/m(2) daily) and after radiotherapy (150 200mg/m(2), for 5 days every 4 weeks). At time of recurrence/progression, a nitrosourea-based chemotherapy constitutes a reasonable option, as well as a temozolomide re-challenge for patients without progression during prior temozolomide treatment. PMID- 18394917 TI - Cognitive functions and elderly cancer patients receiving anticancer treatment: a prospective study. AB - It has been reported that anticancer treatment may cause cognitive impairment. Elderly patients in particular could be at increased risk for treatment-related cognitive deterioration. A consecutive series of cancer out-patients >or=65 years old were prospectively assessed by means of a neuropsychological test Cambridge Cognitive Examination (CAMCOG) test at baseline, and after 3 and 6 months from study entry. Patients were categorized in three groups (group 1, no anticancer treatment; group 2, receiving chemotherapy; group 3, receiving endocrine therapy). Comprehensive geriatric assessment was performed at the three time points evaluation. Sixty-one patients were enrolled (32, 16 and 13, in groups 1, 2, and 3, respectively). At baseline, cognitive function was directly correlated to Activities of Daily Living (ADL) and Instrumental Activities of Daily Living (IADL) scores and was associated with higher educational level and absence of depression. Overall, cognitive function did not worsen across time in each group. However, more patients in the CT group showed worsening in memory skills, and more patients in the ET and CT group experienced reduction in the attention score. PMID- 18394918 TI - Breast cancer and pregnancy: challenges of chemotherapy. AB - BACKGROUND: Breast cancer is the second most frequently occurring malignancy during pregnancy. As evidence-based data on diagnostics and treatment is lacking, current recommendations mostly derive from nonrandomized experiences. We reviewed the current literature with focus on chemotherapy during pregnancy and lactation. RESULTS: The diagnosis of pregnancy associated breast cancer implies the challenge to balance between a life-saving therapy for the mother's breast cancer and a potentially life-threatening therapy for the fetus. With few limitations, surgery and chemotherapy can be performed during pregnancy, preferably in the second and third trimester, whereas radiotherapy and endocrine or antibody treatment should be postponed until after delivery. CONCLUSION: Breast cancer during pregnancy and lactation remains a therapeutic and ethical multidisciplinary challenge. Close cooperation between all disciplines is inevitable to find an optimal treatment strategy for the mother and her unborn child. PMID- 18394919 TI - Dissociation of ACTH and glucocorticoids. AB - It is increasingly clear that significant differential regulation of pituitary and adrenal gland activation exists, leading to a dissociation of plasma adrenocorticotropic hormone and corticosteroid secretion during fetal, postnatal and adult life. An increasing number of preclinical and clinical studies report dissociation of adrenocorticotropic hormone and cortisol levels in critical illness, inflammation and mental disorders. Mechanisms involve an altered adrenal sensitivity, aberrant receptor expression or modulation of adrenal function by cytokines, vasoactive factors or neuropeptides. The degree of dissociation has been associated with the level of complications of sepsis, surgery, malignant disease and depression. The separation of adrenocorticotropic hormone and corticosteroid secretion is of clinical relevance and should be incorporated into our view on endocrine stress regulation. PMID- 18394920 TI - Use of 18O labels to monitor deamidation during protein and peptide sample processing. AB - Nonenzymatic deamidation of asparagine residues in proteins generates aspartyl (Asp) and isoaspartyl (isoAsp) residues via a succinimide intermediate in a neutral or basic environment. Electron capture dissociation (ECD) can differentiate and quantify the relative abundance of these isomeric products in the deamidated proteins. This method requires the proteins to be digested, usually by trypsin, into peptides that are amenable to ECD. ECD of these peptides can produce diagnostic ions for each isomer; the c. + 58 and z - 57 fragment ions for the isoAsp residue and the fragment ion ((M + nH)((n-1)+.) - 60) corresponding to the side-chain loss from the Asp residue. However, deamidation can also occur as an artifact during sample preparation, particularly when using typical tryptic digestion protocols. With 18O labeling, it is possible to differentiate deamidation occurring during trypsin digestion which causes a +3 Da (18O1 + 1D) mass shift from the pre-existing deamidation, which leads to a +1-Da mass shift. This paper demonstrates the use of (18)O labeling to monitor three rapidly deamidating peptides released from proteins (calmodulin, ribonuclease A, and lysozyme) during the time course of trypsin digestion processes, and shows that the fast (approximately 4 h) trypsin digestion process generates no additional detectable peptide deamidations. PMID- 18394921 TI - Know yourself and you shall know the other... to a certain extent: multiple paths of influence of self-reflection on mindreading. AB - Social and neurocognitive research suggests that thinking about one's own thinking and thinking about the thinking of others-termed 'mindreading', 'metacognition', 'social cognition' or 'mentalizing' are not identical activities. The ability though to think about thinking in the first person is nevertheless related to the ability to think about other's thoughts in the third person. Unclear is how these phenomena influence one another. In this review, we explore how self-reflection and autobiographical memory influence the capacity to think about the thoughts and emotions of others. We review studies suggesting that the more individuals are able to reflect on and retrieve episodes from their life narratives, the more they are likely to grasp others' thoughts and emotions. We discuss evidence supporting this possibility including studies of the neurocognitive bases of empathy and self-awareness and how different aspects of self-reflection may impact on mindreading. We also draw from clinical reports how improved self-reflection may result in a more nuanced mindreading, namely persons suffering from schizophrenia and narcissistic personality disorder. We finally discuss the implications for research and practice and consider whether there are conditions in which the reverse is true, where self-reflection might impair mindreading or in which mindreading may facilitate self-reflection. PMID- 18394922 TI - Assessment of parameter settings for SPM5 spatial normalization of structural MRI data: application to type 2 diabetes. AB - Spatial normalization is the process of standardizing images of different subjects into the same anatomical space. The goal of this work was to assess standard and unified methods in SPM5 for the normalization of structural magnetic resonance imaging (MRI) data acquired in mid-life/elderly subjects with diabetes. In this work, we examined the impact of different parameters (i.e. nonlinear frequency cutoff, nonlinear regularization and nonlinear iterations) on the normalization, in terms of the residual variability. Total entropy was used to assess the residual anatomical variability after spatial normalization in a sample of 14 healthy mid-life/elderly control subjects and 24 mid-life/elderly subjects with type 2 diabetes. Spatial normalization was performed using default settings and by varying a single parameter or a combination of parameters. Descriptive statistics and nonparametric tests were used to examine differences in total entropy. Statistical parametric mapping analyses were performed to evaluate the influence of parameter settings on the spatial normalization. Total entropy results and SPM analyses suggest that the best parameters for the spatial normalization of mid-life/elderly image data to the MNI template, when applying the standard approach, correspond to the default cutoff (25 mm), heavy regularization, and the default number of nonlinear iterations (16). On the other hand, when applying the unified approach, the default parameters were the best for spatial normalization of mid-life/elderly image data to the MNI priors. These findings are relevant for studies of structural brain alterations that may occur in normal aging, chronic medical conditions, neuropsychiatric disorders, and neurodegenerative disorders. PMID- 18394923 TI - Brain activation in discourse comprehension: a 3t fMRI study. AB - To date a very small number of functional neuroimaging studies have specifically examined the effects of story coherence on brain activation using long narratives, a procedure fundamental to the study of global coherence. These studies, however, not only yielded notably divergent results, but also featured a number of caveats. It is the purpose of the present study to try to overcome some of these limitations. A left precuneus/posterior cingulate activation related to global coherence comprehension was in consonance with a part of previous literature. However, our most important results corresponded to left parietal regions (angular gyrus, BA 39), this diverging from the previous studies. Recent developments of the situational models of narrative comprehension could explain all these apparently inconsistent results. According to these, different situation models would be created as a function of the content of the narratives, which would yield in turn different patterns of brain activity. Our data also suggest that the same content might also give place to different situation models as a function of the degree of global coherence achieved by the reader or listener. PMID- 18394925 TI - Reproducibility of brain tissue volumes in longitudinal studies: effects of changes in signal-to-noise ratio and scanner software. AB - It is imperative that users of voxel-based morphometry (VBM) be aware of its reproducibility and the factors which influence results. We assessed the reproducibility of a VBM software package (SPM5) in measuring gray matter (GM) and white matter (WM) volumes from at least two consecutive 3D T1-weighted studies in 64 subjects. Factors investigated were the inter-study interval (ISI: 2.2 h to 124 days), signal-to-noise ratio (SNR: number of image averages (NA)=1 or 2), scanner software version and idle time. SNR was measured by direct estimation of tissue noise (SNR(TN)) and mean intensity in noise-only voxels (SNR(NO)). After the scanner software upgrade, voxel intensity increased 5-fold and WM mean SNR(TN) by 24% (p<0.001). Mean WM and GM volume changes in consecutive studies were near 0% (absolute SD of 7 ml and 10 ml respectively). Studies acquired with original scanner software showed a small (1.6%) mean GM volume increase attributed to SNR(TN) increases in five subjects due to scanner maintenance. GM volumes increased with SNR(TN) across the software upgrade (up to 4.3%; p<0.01) and NA=2 acquisitions (up to 4.1%; p<0.001). GM and WM volumes were independent of ISI when ISI did not encompass the software change. Scanner idle times of >6 h decreased SNR by 7% (p<0.001). SPM5 failed to include visible peripheral GM in only 2 subjects. SNR(TN) increases were greater than SNR(NO) increases across the software upgrade. It was concluded that SNR changes significantly influence SPM5-derived GM volumes. SNR may be influenced by scanner software upgrades and hardware condition and should be routinely assessed in studies of brain volume. PMID- 18394924 TI - Diffuse optical tomography of pain and tactile stimulation: activation in cortical sensory and emotional systems. AB - Using diffuse optical tomography (DOT), we detected activation in the somatosensory cortex and frontal brain areas following tactile (brush) and noxious heat stimulation. Healthy volunteers received stimulation to the dorsum of the right hand. In the somatosensory cortex area, tactile stimulation produced a robust, contralateral to the stimulus, hemodynamic response with a weaker activation on the ipsilateral side. For the same region, noxious thermal stimuli produced bilateral activation of similar intensity that had a prolonged activation with a double peak similar to results that have been reported with functional MRI. Bilateral activation was observed in the frontal areas, oxyhemoglobin changes were positive for brush stimulation while they were initially negative (contralateral) for heat stimulation. These results suggest that based on the temporal and spatial characteristics of the response in the sensory cortex, it is possible to discern painful from mechanical stimulation using DOT. Such ability might have potential applications in a clinical setting in which pain needs to be assessed objectively (e.g., analgesic efficacy, pain responses during surgery). PMID- 18394926 TI - MRI of optic neuritis in a rat model. AB - Neuritis of the optic nerve is one of the most frequent early symptoms of multiple sclerosis. There are only scarce data correlating magnetic resonance imaging (MRI) contrast alterations with the underlying pathology, that is inflammation, demyelination, and axonal damage. Here we studied optic neuritis in a rat model of experimental autoimmune encephalomyelitis by comparing in vivo MRI findings from multiple techniques (T1, T2, proton density, magnetization transfer) to histopathology. We further assessed a breakdown of the blood-brain barrier by using Gd-DTPA and indirectly estimated the intracellular accumulation of calcium as a consequence of axonal damage by using manganese-enhanced MRI. Hyperintensity on T2-weighted images and signal enhancement after Gd-DTPA were highly sensitive to lesions of the optic nerve but did not differentiate between mild, moderate, and severe damage. Signal reduction on T1-weighted images was less sensitive but correlated well with the severity of tissue damage. No significant changes in magnetization transfer ratio were observed. Manganese ions tended to accumulate in the central parts of the inflamed optic nerve. The resulting signal enhancement at 24 h after administration positively correlated with the severity of axonal loss. Thus, manganese might be an indicator of intracellular calcium accumulation that is known to be associated with axon damage. Although none of the methods alone distinguished between inflammation, demyelination, and reduced axon density, their specific capabilities should prove useful for future in vivo MRI studies of optic neuritis in both animal models and humans. PMID- 18394927 TI - Analyzing information flow in brain networks with nonparametric Granger causality. AB - Multielectrode neurophysiological recording and high-resolution neuroimaging generate multivariate data that are the basis for understanding the patterns of neural interactions. How to extract directions of information flow in brain networks from these data remains a key challenge. Research over the last few years has identified Granger causality as a statistically principled technique to furnish this capability. The estimation of Granger causality currently requires autoregressive modeling of neural data. Here, we propose a nonparametric approach based on widely used Fourier and wavelet transforms to estimate both pairwise and conditional measures of Granger causality, eliminating the need of explicit autoregressive data modeling. We demonstrate the effectiveness of this approach by applying it to synthetic data generated by network models with known connectivity and to local field potentials recorded from monkeys performing a sensorimotor task. PMID- 18394929 TI - Unraveling the evolutionary history of the hyperdiverse ant genus Pheidole (Hymenoptera: Formicidae). AB - Pheidole is currently the most species rich genus of ants in the world, with many taxa still awaiting description. In this study, I reconstruct the phylogeny of Pheidole using molecular characters from three mitochondrial genes and two nuclear genes for approximately 140 species. The phylogenetic relationships of Pheidole are investigated with special interest in understanding factors that may have led to their remarkable diversity. The results presented here establish a framework for understanding the explosive radiation of this group by providing (1) a phylogenetic estimate, and (2) a comparative analysis of life history traits that are likely to have been important in the diversification of the group. In all analyses, Pheidole is recovered as a monophyletic lineage, and molecular clock estimates infer an age of 58.4-61.2 million years ago (Ma) for crown group members of the genus. Using an estimate of diversification rate, it appears that Pheidole has undergone 0.108-0.103 speciation events per million years. Previous hypotheses of species groups were largely not upheld in the analyses presented here. Workers of the genus Pheidole are dimorphic with a minor and major (soldier) subcaste. A third subcaste of super majors is known in eight species of Pheidole and this trait was found to have arisen multiple times throughout the phylogeny. Seed harvesting is common among species of the genus and is thought to be one of the factors leading to the diversification of the group, but additional data will be required to further test this hypothesis. To address biogeographic questions on the origin of the genus, both New and Old World species were included in these analyses, and the results suggest that Pheidole is New World in origin with a possible single introduction into the Old World. PMID- 18394928 TI - Segmentation of age-related white matter changes in a clinical multi-center study. AB - Age-related white matter changes (WMC) are thought to be a marker of vascular pathology, and have been associated with motor and cognitive deficits. In the present study, an optimized artificial neural network was used as an automatic segmentation method to produce probabilistic maps of WMC in a clinical multi center study. The neural network uses information from T1- and T2-weighted and fluid attenuation inversion recovery (FLAIR) magnetic resonance (MR) scans, neighboring voxels and spatial location. Generalizability of the neural network was optimized by including the Optimal Brain Damage (OBD) pruning method in the training stage. Six optimized neural networks were produced to investigate the impact of different input information on WMC segmentation. The automatic segmentation method was applied to MR scans of 362 non-demented elderly subjects from 11 centers in the European multi-center study Leukoaraiosis And Disability (LADIS). Semi-manually delineated WMC were used for validating the segmentation produced by the neural networks. The neural network segmentation demonstrated high consistency between subjects and centers, making it a promising technique for large studies. For WMC volumes less than 10 ml, an increasing discrepancy between semi-manual and neural network segmentation was observed using the similarity index (SI) measure. The use of all three image modalities significantly improved cross-center generalizability compared to neural networks using the FLAIR image only. Expert knowledge not available to the neural networks was a minor source of discrepancy, while variation in MR scan quality constituted the largest source of error. PMID- 18394930 TI - Nutritional risk factors for the development of hypertension in diabetic patients. AB - The aim of this cross-sectional study was to determine the significant limitations, sensitivity, specificity, partial correlations, and odds ratios of nutrient intake in patients with and without hypertension with Type 2 diabetes mellitus. Diabetic patients (n=220) with clinical diagnosis of hypertension and diabetic patients (n=230) without hypertension were included in this study. The questionnaire form included a list of 65 food items formed from five main food groups (grain, meat and alternatives, dairy products, vegetables-fruits and fat) and 25 dietary habits. When both groups were compared and analyzed by logistic regression, black tea consumption (OR=0.823, P<.001), vegetables-fruits scores (OR=0.853, P<.001), triglycerides (OR=0.726, P<.05), waist-to-hip ratio (WHR) (OR=0.777, P<.01) and high-density lipoprotein cholesterol (HDL-C) (OR=0.526, P<.001) made significant differences. In ROC curves, the area under the curve of black tea (0.921), vegetables-fruits (0.906), triglycerides (0.889), WHR (0.881) and HDL-C (0.820) provided high accuracy to distinguish between patients with and without hypertension (P<.001). In diabetic patients without hypertension, significant partial correlations were observed between blood pressure and dairy products (systolic: r=)0.14; diastolic: r=)0.14, P<.05), vegetables-fruits groups (systolic: r=)0.18; diastolic: r=)0.17, P<.01) and black tea intake (systolic: r=)0.23; diastolic: r=)0.22, P<.001). It has been found that higher intake of black tea and vegetables-fruits consumption in diabetic patients protect against developing hypertension. PMID- 18394931 TI - Normal insulin response to short-term intense exercise is abolished in Type 2 diabetic patients treated with gliclazide. AB - BACKGROUND: Physical activity is an essential component of diabetes management; however, exercise is associated with the risk for metabolic decompensation. The aim of the study was to analyze insulin response to the short-term intense exercise in middle-aged Type 2 diabetic patients treated with gliclazide. MATERIALS AND METHODS: Fourteen Type 2 diabetic patients (47.9+/-1.6 years, mean+/-S.E.M.), treated with gliclazide, and 14 healthy controls (45.1+/-1.0 years) were submitted to standard graduated submaximal (90% HR(max)) exercise treadmill testing at 2 h after standardized breakfast. Serum glucose, insulin, proinsulin, C peptide, growth hormone, insulin-like growth factor-1, and cortisol concentrations; and plasma lactate, glucagon, epinephrine, and norepinephrine concentrations were measured during the periexercise period up to the 60th min of the recovery period. RESULTS: Significant hemodynamic (heart rate, systolic, and diastolic blood pressure), metabolic (lactate concentration), and hormonal (epinephrine and norepinephrine levels) responses to the exercise were similar in patients and healthy subjects. Glucose, insulin, and proinsulin levels were higher in the diabetic group at the preexercise and at all the next analyzed time points. The insulin concentration increased during the postprandial period in both groups and decreased subsequently during the exercise only in the control group, without concurrent C peptide decline. The C peptide-to-insulin ratio increased during the exercise and decreased immediately postexercise only in the control group. CONCLUSIONS: The initial decrease of the insulin serum concentration during short-term intense exercise in normal middle-aged men is primarily related to the increased clearance of the hormone. Normal insulin response to the exercise was abolished in Type 2 diabetic patients treated with gliclazide. PMID- 18394932 TI - Fifty percent area reduction after 4 weeks of treatment is a reliable indicator for healing--analysis of a single-center cohort of 704 diabetic patients. AB - INTRODUCTION: The aim of the study was to investigate whether an area reduction greater than 50% within the first 4 weeks of treatment is associated with a higher long-term probability of healing. PATIENTS AND METHODS: We treated diabetic foot ulcers according to a comprehensive interdisciplinary wound care protocol. Follow-up was documented through a special wound documentation system. Data were entered into SPSS for statistical analysis to calculate the probability of healing according to the Kaplan-Meier method. Results were expressed as median (minimum-maximum), and the percentage of area reduction (PA) was defined as [(area(4 weeks)/area(baseline))x100)/area(baseline). Patients were divided into responders when PA reached at least 50% and nonresponders when PA was less than 50%. Healing was defined as PA=100%. RESULTS: In total, 704 patients were included into the analysis. Median time of follow-up was 71 (2-365) days. Wound duration was 31 (1-4018) days, and the initial wound size was calculated to be 1.18 (0.1-99) cm(2). In 27.8%, there was a positive probing to bone; in 64.5%, both pedal pulses were not palpable. Major amputation rate was 2.8% and minor amputation rate was 10.2%. The overall probability of healing was 35% after 12 weeks, 41% after 16 weeks, and 73% after 1 year. The surrogate visit (4 weeks) was performed after a median of 27 (14-42) days without a difference between responders and nonresponders. There were 334 (47%) responders and 370 (53%) nonresponders. Responders had a significantly higher probability of healing compared with nonresponders (12 weeks: 52.3% vs. 18.4%, P=.0001; 16 weeks: 46.7% vs. 26.5%, P=.0001; 1 year: 82.5% vs. 64.9%, P=.0001). CONCLUSIONS: The calculation of the percentage of area reduction after 4 weeks of treatment is a valid tool to estimate the probability of healing. In clinical practice, a reevaluation of the treatment schedule is recommended for wounds that do not reach 50% area reduction within the first 4 weeks of therapy. PMID- 18394933 TI - Cardioprotective effect of vitamin E: rescues of diabetes-induced cardiac malfunction, oxidative stress, and apoptosis in rat. AB - AIM: This study was designed to assess the effect of vitamin E on cardiac autonomic neuropathy, cardiomyocyte apoptosis, and the status of oxidative stress in the heart under hyperglycemic conditions, in vivo. METHODS: Wistar male rats (n=16) were made hyperglycemic by streptozotocin at 6 months of age. Normal Wistar rats (n=8) of the same age were used as the control group. Diabetic rats were divided into two groups, nontreated and those treated with vitamin E (300 mg/day). Stable hyperglycemic status was proved by levels of blood sugar and HbA(1c). Lipid peroxidation, protein oxidation, and cellular antioxidant defense were measured by 8-isoprotane, protein carbonyl content, and superoxide dismutase (SOD) activity, respectively. RESULTS: Cardiac complications such as autonomic neuropathy as prolonged QT interval along with significant increases in level of 8-isoprotane, protein carbonyl content, and SOD activity were observed after 6 weeks. Structural abnormality was also observed as severe induction of apoptosis in cardiomyocytes. CONCLUSION: Significant decline in apoptosis, lipid peroxidation, protein oxidation, and QT interval resulted from vitamin E administration, which strongly implies that this radical scavenger may promote a convalescing effect on diabetic cardiomyopathy through the attenuation of oxidative stress and abrogation of apoptotic signals, which was verified by restoring normal QT interval. PMID- 18394935 TI - Growth factor contents of autologous human sera prepared by different production methods and their biological effects on chondrocytes. AB - To discuss the autologous serum production for cartilage tissue engineering, we compared three kinds of sera: whole blood-derived serum (WBS), platelet containing plasma-derived serum (PCS), and plasma-derived serum (PDS), on the growth factor contents and their biological effects on human auricular chondrocytes. EGF, VEGF and PDGF levels were highest in WBS, while PCS and PDS followed WBS. The proliferation effects of WBS were the most pronounced, followed by that of PCS, both of which realized a 1000-fold-increase in chondrocyte numbers at the third passage, whereas PDS reached it after passage 4. No significant differences were observed in histology or cartilaginous matrix measurements of tissue-engineered cartilage produced from chondrocytes cultured under different serum conditions. WBS would be clinically useful because of its potent proliferation effects, while PCS, which possibly saves the red cell concentrate, may be an option in cases where there are elevated risks of blood loss. PMID- 18394937 TI - Linear phase slope in pulse design: application to coherence transfer. AB - Using optimal control methods, robust broadband excitation pulses can be designed with a defined linear phase dispersion. Applications include increased bandwidth for a given pulse length compared to equivalent pulses requiring no phase correction, selective pulses, and pulses that mitigate the effects of relaxation. This also makes it possible to create pulses that are equivalent to ideal hard pulses followed by an effective evolution period. For example, in applications, where the excitation pulse is followed by a constant delay, e.g. for the evolution of heteronuclear couplings, part of the pulse duration can be absorbed in existing delays, significantly reducing the time overhead of long, highly robust pulses. We refer to the class of such excitation pulses with a defined linear phase dispersion as ICEBERG pulses (Inherent Coherence Evolution optimized Broadband Excitation Resulting in constant phase Gradients). A systematic study of the dependence of the excitation efficiency on the phase dispersion of the excitation pulses is presented, which reveals surprising opportunities for improved pulse sequence performance. PMID- 18394938 TI - Capillary scale NMR flow mapping. AB - Stopped-flow NMR at capillary scale has many advantages over traditional methods of introducing the sample into the probe, particularly when large numbers of samples must be examined. This work describes application of a simple method for direct visualization of a sample inside the flow cell of flow NMR systems to capillary scale analysis. We describe the details of the method and show how it can be used to measure the optimum flow rate for a capillary NMR system and how to determine the optimum sampling efficiency for small samples. PMID- 18394936 TI - NKG2D-deficient mice are defective in tumor surveillance in models of spontaneous malignancy. AB - Ligands for the NKG2D stimulatory receptor are frequently upregulated on tumor lines, rendering them sensitive to natural killer (NK) cells, but the role of NKG2D in tumor surveillance has not been addressed in spontaneous cancer models. Here, we provided the first characterization of NKG2D-deficient mice, including evidence that NKG2D was not necessary for NK cell development but was critical for immunosurveillance of epithelial and lymphoid malignancies in two transgenic models of de novo tumorigenesis. In both models, we detected NKG2D ligands on the tumor cell surface ex vivo, providing needed evidence for ligand expression by primary tumors. In a prostate cancer model, aggressive tumors arising in NKG2D deficient mice expressed higher amounts of NKG2D ligands than did similar tumors in wild-type mice, suggesting an NKG2D-dependent immunoediting of tumors in this model. These findings provide important genetic evidence for surveillance of primary tumors by an NK receptor. PMID- 18394939 TI - Molecular mechanics and molecular orbital simulations on specific interactions between peroxisome proliferator-activated receptor PPARalpha and plasticizer. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) has various physiological functions such as lipid and glucose metabolism, inflammation and fibrosis in living organisms. Many types of ligand molecules such as phthalate and adipate esters control these physiological functions. In the present study, to elucidate the dependence of PPARalpha properties on ligand binding, we investigated stable structures and electronic properties for the complexes of PPARalpha and phthalate as well as adipate esters, which are used as a plasticizer, by molecular simulations based on molecular mechanics and molecular orbital methods. Furthermore, to elucidate the influence of these esters in vivo, we injected them into male mice and observed the change in the expression of PPARalpha-related enzymes. The comparison between the calculated and observed results indicates that the change in the expression has a correlation with the size of energy gaps between highest occupied and lowest unoccupied molecular orbitals of the complexes with mouse PPARalpha and esters. PMID- 18394940 TI - Effects of neuraminidase on equine isolated bronchi. AB - It has been demonstrated in mammals that the airway hyper-responsiveness, which accompanies viral infections, is the result of increased reflex bronchoconstriction due to inhibition of muscarinic prejunctional receptors, which belong to M2 subtypes. Multiple mechanisms account for virus-induced M2 receptor dysfunction. Viral neuraminidase may deglycosylate the M2 receptor, decreasing acetylcholine affinity. Equine influenza remains a common viral respiratory disease of horses worldwide, which results in loss to the equine industry, by decreasing performance, convalescence time and loss of peak performance due to chronic sequelae, such as airway hyper-responsiveness. The purpose of this study was to evaluate the effect of neuraminidase on equine isolated bronchi, assessed in equine bronchial smooth muscle rings, derived from five healthy equine male lungs. A pretreatment with vehicle did not modify contraction induced by electrical field stimulation (EFS) studies at each frequency tested. A pretreatment with pilocarpine (1-100 microM) significantly reduced, while methoctramine (1-100 microM) significantly increased contraction induced by EFS. Finally, neuraminidase (0.5 UI) significantly increased contraction induced by EFS. These results suggest that airway hyper responsiveness that follows a viral influenza infection might be related to a dysfunction of muscarinic prejunctional receptors. PMID- 18394941 TI - Influence of temperature acclimation and gut content on the supercooling ability of the land snail Cornu aspersum. AB - The invasive land snail Cornu aspersum possesses a low ability to supercool (c. 5 degrees C in winter) and survives only minimal ice formation in its body fluids, what may limit its expansion to colder environments. In the present study, we investigated the influence of acclimation and starvation on its supercooling ability. During eight weeks, individuals were maintained at 20 degrees C, fed or starved, or placed at 5 degrees C, directly or with progressive acclimation to cold and shorter photoperiod. Temperature of crystallisation of whole individual (Tc(I)) and hemolymph (Tc(H)), mass data and gut content were recorded every two weeks. Hemolymphatic glucose and glycerol were measured at the end of experiment and occurrence of intestinal ice-nucleating agents (INA) was researched. Acclimation had no effect on Tc(I) but stimulated purging of the gut. Starvation induced a slight decrease of Tc(I) whereas a high quantity of alimentary particles in the digestive tract limited the supercooling ability. Glucose and glycerol were not synthesized in cold conditions. Mean Tc(H) was low (c. -17 degrees C), some INA being present in hemolymph of fed animals. Intestinal content of starved individuals exhibited a mean Tc of c. -6 degrees C, decreasing to c. -12 degrees after heating, suggesting the presence of organic INA. PMID- 18394942 TI - Isolation and partial characterization of three isoamylases of Rhyzopertha dominica F. (Coleoptera: Bostrichidae). AB - Three isoamylases of Rhyzopertha dominica (termed RdA70, RdA79, and RdA90 according to their relative mobility in gel electrophoresis) were isolated by ammonium sulfate fractionation and hydrophobic interaction chromatography. RdA70 and RdA79 showed an optimal pH of 7.0, whereas for RdA90 the optimal pH was 6.5. The three isoamylases remained stable at 50 degrees C for 1 h, but at 60 degrees C, all lost 50% of their activity in 20 min and were completely inactivated in 1 h. RdA70 and RdA79 were inhibited by albumin extracts from wheat samples varying widely in amylase inhibitory activity; however, RdA90 was highly resistant to inhibition. beta-Mercaptoethanol up to 30 mM increased the activity of the three isoamylases by 2.5-fold. The action pattern of the three isoamylases was typical of endoamylases; however, differences were observed on the hydrolytic efficiency rates measured as V(max)/K(m) ratio on starch, amylopectin, and amylose. The hydrolyzing action of RdA90 on starch and amylopectin (V(max)/K(m)=90.4+/-2.3 and 78.9+/-6.6, respectively) was less efficient than that on amylose (V(max)/K(m)=214+/-23.2). RdA79 efficiently hydrolyzed both amylopectin and amylose (V(max)/K(m)=260.6+/-12.9 and 326.5+/-9.4, respectively). RdA70 hydrolyzed starch and amylose at similar rates (V(max)/K(m)=202.9+/-5.5 and 215.9+/-6.2, respectively), but amylopectin was a poor substrate (V(max)/K(m)=124.2+/-7.4). The overall results suggest that RdA70 and RdA79 appear to belong to a group of saccharifying isoamylases that breaks down long fragments of oligosaccharide chains produced by the hydrolytic action of RdA90. The simultaneous action of the three isoamylases on starch, aside from the high resistance of RdA90 to wheat amylase inhibitors, might allow R. dominica to feed and reproduce successfully on the wheat kernel. PMID- 18394943 TI - Frequency and associated factors of proteinuria in Iranian HIV-positive patients. AB - BACKGROUND: Screening HIV-positive patients for proteinuria would result in early recognition of HIV-associated nephropathy (HIVAN). This would allow diagnosis and treatment of HIVAN at an early stage and hence prevent further disease progression. This study was undertaken to determine the frequency of proteinuria and its associated factors in Iranian HIV-positive patients. METHODS: In this study, 171 HIV-positive patients were screened for proteinuria. Proteinuria was defined as > or =1+ protein on the urine dipstick. A questionnaire was used to collect patient sociodemographic and clinical data. Hepatitis B surface antigen (HBsAg), hepatitis C antibody (anti-HCV), serum albumin, and creatinine were tested in all patients. CD4 counts were obtained by flow cytometry. RESULTS: Out of 171 HIV-positive patients, 21 (12.3%) had proteinuria. There were no significant differences between patients with and without proteinuria with regard to age, sex, risk behaviors for HIV acquisition, stage of infection, concurrent antiretroviral therapy, systolic and diastolic blood pressure, serum albumin and creatinine, glomerular filtration rate (GFR), and presence of anti-HCV or HBsAg. Patients with proteinuria had a lower CD4 count and creatinine clearance than those without proteinuria. CONCLUSION: Proteinuria was relatively high in Iranian HIV-positive patients. The group at higher risk was that of patients with lower CD4 counts and creatinine clearance. PMID- 18394944 TI - Congenital spinal tuberculosis associated with asymptomatic endometrial tuberculosis: A rare case report. AB - Spinal tuberculosis constitutes 50% of all musculoskeletal tuberculosis. However, literature regarding congenital spinal tuberculosis is very scanty. Congenital spinal tuberculosis was diagnosed in a two-month-old child on the basis of age at presentation (gibbus since three weeks of age), hepatomegaly, raised ESR, radiological destruction of D10-D11 vertebrae, asymptomatic maternal endometrial tuberculosis and tuberculous histopathology from CT guided biopsy specimen from D10 vertebra. Both child and mother were treated by antitubercular treatment. Child improved symptomatically, gibbus became less prominent and ESR became normal at the end of one year of treatment. Patient had no recurrence during last two years of follow-up. This is the first case of congenital tuberculosis of spine with a documented source of infection from asymptomatic maternal endometrial tuberculosis. CT guided core biopsy from vertebra/aspiration from the paravertebral abscess help in early diagnosis and treatment to reduce neurological morbidity and mortality. Endometrial biopsy help in establishing the diagnosis of congenital tuberculosis and adequate antitubercular treatment in such cases may provide protection to fetus in subsequent pregnancy. PMID- 18394945 TI - Circulating adrenomedullin levels in ankylosing spondylitis and Familial Mediterranean Fever. AB - INTRODUCTION: Adrenomedullin (AM) is a 52-amino acid peptide with vasorelaxant properties. Apart from its roles on vascular tonus, AM can also contribute to inflammatory events. Plasma AM levels were elevated in connective tissue diseases and vasculitic disorders. Ankylosing spondylitis (AS) is a chronic inflammatory disease of the spine initiating in the sacroiliac joints. Familial Mediterranean Fever (FMF) is a hereditary disorder characterized by self-limiting acute attacks of fever and the presence of sustained subclinical inflammation in the attack free periods. In this study, we investigated plasma AM levels in patients with AS and patients with FMF. METHODS: Twenty AS patients with active disease manifestations (mean age: 41.6+/-10.9 years, female/male: 7/13), 28 FMF patients with acute attack (mean age: 27.4+/-10.7 years, female/male: 17/11), and 26 healthy controls (mean age: 39.9+/-5.5 years, female/male: 16/10) were enrolled in this study. AM levels were also measured in 11 FMF patients 2 months after the cessation of their attacks. AM levels of those 11 patients during their FMF attacks and attack-free periods were also compared. RESULTS: Median plasma AM levels were 23.86 (17.24-40.09) pmol/mL, 27.33 (17.24-38.52) pmol/mL, and 26.11 (17.05-37.42) pmol/mL in AS patients, FMF patients with acute attack, and healthy controls, respectively (p>0.05). AM levels were also similar in the attack-free periods of FMF patients [26.35 (24.35-34.14) pmol/mL]. There was no correlation between plasma AM levels and C-reactive protein, or between plasma AM levels and erythrocyte sedimentation rate. CONCLUSIONS: AM does not seem to have any role in the pathogenesis of AS and FMF. Previous reports of elevated levels of AM in connective tissue disorders and vasculitic diseases are probably disease specific, and AM does not seem to be a common component of inflammatory rheumatic disorders. PMID- 18394946 TI - [Offer external cephalic version, you will avoid breech deliveries!]. PMID- 18394947 TI - The solvent-free synthesis of 1,4-dihydropyridines under ultrasound irradiation without catalyst. AB - The condensation of aldehydes, ethyl acetoacetate and ammonium acetate result 1,4 dihydropyridines in 82-99% yields under ultrasound irradiation without solvent and catalyst at room temperature. Compared with conventional methods, the main advantages of the present procedure are milder conditions, shorter reaction time and higher yields. PMID- 18394949 TI - Grey matter atrophy in early versus late dementia in Parkinson's disease. AB - There is considerable heterogeneity within patients with Parkinson's disease (PD) regarding time from onset of PD to dementia. We compared grey matter atrophy using magnetic resonance images (MRIs) of patients with PD who developed dementia early and late in the disease course. The group with early dementia had more atrophy than the group with late dementia in some areas, while the late dementia group had symmetrical reduction in concentration of grey matter in the insula bilaterally compared to the early dementia group. Our results indicate that the early development of dementia in PD is associated with more severe degeneration of cortical and sub-cortical structures. PMID- 18394950 TI - Resistance against novel anticancer metal compounds: differences and similarities. AB - The platinum antitumor drugs cisplatin, carboplatin and oxaliplatin are widely used components of modern cancer chemotherapy. However, their success is limited by severe adverse effects and because of the impact of intrinsic and acquired resistance mechanisms on tumor responses. Consequently, intense efforts have been made to develop new metal compounds that either display enhanced tumor specificity or are less prone to the development of resistance. Despite the synthesis of thousands of compounds during the last decades only very few novel metal drugs have successfully reached clinical development and/or approval so far. In this review we summarize the current knowledge on drug resistance mechanisms against novel metal compounds (including platinum, arsenic, ruthenium, gallium, titanium, copper, and lanthanum drugs), and address the question whether there might exist a general metal-drug resistance phenotype. PMID- 18394951 TI - Identification and characterization of Paragonimus westermani leucine aminopeptidase. AB - Paragonimus westermani is a tissue-invading trematode parasite that causes inflammatory lung disease as well as systemic infections including cerebral invasion in carnivorous mammals. While aminopeptidases play important roles in trematodes in the catabolism of host hemoglobin, an essential source of nutrient for the parasite, little is known about aminopeptidase in Paragonimus. Presently, we isolated a cDNA encoding a 58 kDa P. westermani leucine aminopeptidase (PwLAP). Deduced amino acid sequence of PwLAP exhibited significant sequence homology with LAP from Schistosoma spp. and Fasciola hepatica. Biochemical analysis of the recombinant PwLAP protein demonstrated preferential substrate specificity for Leu-NHMec and inhibition by EDTA, 1,10-phenanthroline, and bestatin, which are conserved characteristics of the M17 family of leucine aminopeptidase. PwLAP exhibited relatively higher enzyme activity in the presence of Mn2+ compared to Schistosoma mansoni LAP. Based on the biochemical properties and immunohistochemical analysis, PwLAP is concluded to represent a leucine aminopeptidase. The enzyme is most likely responsible for the catabolism of host hemoglobin, and, hence, represents a potential target of Paragonimus chemotherapy. PMID- 18394952 TI - Novel mechanistic aspects on the reaction between low spin Fe(II) Schiff base amino acid complexes and hydrogen peroxide-spectrophotometric tracer of intraperoxo intermediate catalyzed reaction. AB - The kinetics and mechanism of the reaction of hydrogen peroxide with some Fe(II) Schiff base complexes were investigated spectrophotometrically in aqueous solution at pH 8 and 35 degrees C under pseudo-first-order conditions. The used ligands were derived from salicylaldehyde or o-hydroxynaphthaldehyde and some amino acids (l-leucine, l-iso-leucine, l-serine, l-methionine and dl-tryptophan). It was found that the formation of the purple interaperoxo complex appears only above pH 7.5. The reaction consists of two steps. The first step involves reversible formation of the intraperoxo intermediate which renders irreversible at pH 8. The second step consists of inner-sphere electron transfer. The suggested scheme illustrated first-order kinetics at low [H(2)O(2)] and zero order at high [H(2)O(2)]. Moreover, the activation parameters of the reaction were evaluated. PMID- 18394953 TI - ESR, electrochemical and cyclodextrin-inclusion studies of triazolopyridyl pyridyl ketones and dipyridyl ketones derivatives. AB - The electron spin resonance (ESR) spectra of free radicals obtained by electrolytic reduction of triazolopyridyl pyridyl ketones and dipyridyl ketones derivatives were measured in dimethylsulfoxide (DMSO). The hyperfine patterns indicate that the spin density delocalization is dependent of the rings presented in the molecule. The electrochemistry of these compounds was characterized using cyclic voltammetry, in DMSO as solvent. When one carbonyl is present in the molecule one step in the reduction mechanism was observed while two carbonyl are present two steps were detected. The first wave was assigned to the generation of the correspondent free radical species, and the second wave was assigned to the dianion derivatives. The phase-solubility measurements indicated an interaction between molecules selected and cyclodextrins in water. These inclusion complexes are 1:1 with betaCD, and HP-betaCD. The values of Ks showed a different kind of complexes depending on which rings are included. AM1 and DFT calculations were performed to obtain the optimized geometries, theoretical hyperfine constants, and spin distributions, respectively. The theoretical results are in complete agreement with the experimental ones. PMID- 18394954 TI - Genetic diversity of the Kaposi's sarcoma herpesvirus K1 protein in AIDS-KS in Zimbabwe. AB - BACKGROUND: Kaposi's sarcoma-associated herpesvirus (KSHV) encodes genetically diverse K1 alleles which have unique geographic distributions. Little is known about K1 genetic diversity in Zimbabwe where acquired immunodeficiency syndrome associated KS (AIDS-KS) is epidemic. OBJECTIVE: Evaluate K1 diversity in Zimbabwe and compare Zimbabwean K1 diversity to other areas in Africa. STUDY DESIGN: K1 nucleotide sequence was determined for AIDS-KS cases in Zimbabwe. K1 references sequences were obtained from Genbank. RESULTS: Among 65 Zimbabwean AIDS-KS cases, 26 (40%) were K1 subtype A and 39 (60%) were subtype B. Zimbabwean subtype A sequences grouped only with African intratype A5 variants. Zimbabwean subtype B sequences grouped with multiple intratype African variants: 26 B1 (26%), four B3 (6%) and nine highly divergent B4 (14%). Zimbabwean subtype B had a lower synonymous to nonsynonymous mutation ratio (median 0.59 versus 0.66; P=0.008) and greater distance to the most recent common ancestor (median 0.03 versus 0.009; P<0.001) compared to subtype A. Within the B subgroup, the distribution of intratype B variants differed in Zimbabwe and Uganda (P=0.004). CONCLUSIONS: Greater positive selection and genetic diversity in K1 subtype B compared to subtype A5 exist in Zimbabwe. However, there were no significant associations between K1 subtype and the clinical or demographic characteristics of AIDS-KS cases. PMID- 18394955 TI - Evaluation of the Loopamp (loop-mediated isothermal amplification) kit for detecting Norovirus RNA in faecal samples. AB - BACKGROUND: Noroviruses (NoVs) are associated with outbreaks of diarrhoeal illness in hospitals, nursing and residential homes and other institutional settings. NoV strains exhibit wide genetic diversity, and different virus genogroups and genotypes co-circulate in any geographical region at the same time, although most outbreaks of gastroenteritis are predominantly associated with genogroup II. The reverse transcription-polymerase chain reaction (RT-PCR) is the gold standard for detecting NoVs in clinical samples. OBJECTIVES: This study evaluates commercialised Loopamp kits for detecting NoV GI and NoV GII in faecal samples collected from patients with gastroenteritis and compares the results with those obtained using real-time RT-PCR with NoV genogroup sequence specific detection. STUDY DESIGN: Five hundred and ten faecal samples collected from patients with gastroenteritis were evaluated for the presence of NoV using the gold-standard real-time RT-PCRs and the Loopamp assays. RESULTS: The Loopamp Norovirus GI and GII detection kits performed well compared to genogroup-specific real-time RT-PCR. Although the sensitivity of detection of GI strains (83.3%) was less than that for GII strains (97.4%), this will have little impact on the laboratory diagnosis of NoV, since GII strains are associated with the majority of outbreaks examined. CONCLUSIONS: The Loopamp GII detection kit is a sensitive method for detecting all the commonly circulating GII-4 strains included in the evaluation panel. PMID- 18394956 TI - Time-locked association between rapid cerebral blood flow modulation and attentional performance. AB - OBJECTIVE: The study investigated relationships between rapid cerebral hemodynamic modulation and attentional performance. Based on former results on complex cognitive functioning, a specific association between the first seconds of the hemodynamic response and performance was hypothesized. METHODS: Using transcranial Doppler sonography, blood flow velocities in the middle cerebral arteries of both hemispheres were recorded in 48 healthy subjects. The applied task comprised motor reactions on a visual stimulus which was preceded by an acoustic warning signal (interstimulus interval 5s). Task-induced hemodynamic changes were assessed second-by-second, and related to reaction time using analysis of variance and linear regression. RESULTS: A right dominant blood flow response was observed. Flow velocity increase in the middle fraction of the interstimulus interval, i.e. seconds 2 and 3 after the cuing signal, significantly correlated with reaction time. This was not the case for the very early and late components of the response. CONCLUSIONS: The results suggest a time-locked association between cerebral blood flow increase and attentional performance. This is in accordance with neurophysiological studies that revealed the closest relationship between brain perfusion and cortical activity during a similar time window. SIGNIFICANCE: The study supports the assumption of a specific, relatively early time interval in which relationships between cerebral blood flow and behavior become apparent. PMID- 18394957 TI - Evolutionary analysis of synteny and gene fusion for pyrimidine biosynthetic enzymes in Euglenozoa: an extraordinary gap between kinetoplastids and diplonemids. AB - A unique feature of the genome architecture in the parasitic trypanosomatid protists is large-scale synteny. We addressed the evolutionary trait of synteny in the eukaryotic group, Euglenozoa, which consists of euglenoids (earliest branching), diplonemids, and kinetoplastids (trypanosomatids and bodonids). Synteny of the pyrimidine biosynthetic (pyr) gene cluster, which constitutes part of a large syntenic cluster in trypanosomatids and includes four separate genes (pyr1-pyr4) and one fused gene (pyr6/pyr5 fusion), was conserved in the bodonid, Parabodo caudatus. In the diplonemid, Diplonema papillatum, we identified pyr4 and pyr6 genes. Phylogenetic analyses of pyr4 and pyr6 showed the separate origin of each in kinetoplastids and euglenoids/diplonemids and suggested that kinetoplastids have acquired these genes via lateral gene transfer (LGT). Because replacement of genes by non-orthologs within the syntenic cluster is highly unlikely, we concluded that, after separation of the line leading to diplonemids, the syntenic pyr gene cluster was established in the common ancestor of kinetoplastids, preceded by their acquisition via LGT. Notably, we found that diplonemid pyr6 is a stand-alone gene, inconsistent with both euglenoid pyr5/pyr6 and kinetoplastid pyr6/pyr5 fusions. Our findings provide insights into the evolutionary gaps within Euglenozoa and the evolutionary trait of rearrangement of gene fusion in this lineage. PMID- 18394958 TI - Distribution of corazonin and pigment-dispersing factor in the cephalic ganglia of termites. AB - Distribution of neurones detectable with antisera to the corazonin (Crz) and the pigment-dispersing factor (PDF) was mapped in the workers or pseudergates of 10 species representing six out of seven termite families. All species contained two triads of Crz-immunoreactive (Crz-ir) neurones in the protocerebrum. Their fibres were linked to the opposite hemisphere, formed a network in the fronto-lateral protocerebrum, and projected to the corpora cardiaca (CC); in most species the fibres also supplied the deuto- and tritocerebrum and the frontal ganglion. Some species possessed additional Crz-ir perikarya in the protocerebrum and the suboesophageal ganglion (SOG). The PDF-ir somata were primarily located in the optic lobe (OL) and SOG. OL harboured a group (3 groups in Coptotermes) of 2-6 PDF-ir cells with processes extending to the medulla, connecting to the contralateral OL, forming 1-2 networks in the protocerebrum, and in most species running also to CC. Such a PDF-ir system associated with the OL was missing in Reticulitermes. Except for Mastotermes, the termites contained 1-2 PDF-ir cell pairs in the SOG and two species had additional perikarya in the protocerebrum. The results are consistent with the view of a monophyletic termite origin and demonstrate how the Crz-ir and PDF-ir systems diversified in the course of termite phylogeny. PMID- 18394959 TI - First maxillae suction discs in Branchiura (Crustacea): development and evolution in light of the first molecular phylogeny of Branchiura, Pentastomida, and other "Maxillopoda". AB - The fish ectoparasites Branchiura (Crustacea) display two different ways of attachment to the fish surface as adults: the first maxillae are either hooks (Dolops) or suction discs (Argulus, Chonopeltis, and Dipteropeltis). In larval Argulus foliaceus the first maxillae are hooks. With the first molecular phylogeny of the Branchiura as a background, the present paper discusses the evolutionary scenarios leading to hooks versus suction discs. Specific homologies exist between larval Argulus foliaceus hooks and adult Dolops ranarum hooks. These include the presence of a comparable number of segments/portions and a distal segment terminating in a double structure: a distal two-part hook (in Argulus) or one hook and an associate spine-like structure (in Dolops). In the phylogenetic reconstruction based on three molecular markers (mitochondrial 16S rRNA, nuclear 18S and 28S rRNA), Dolops ranarum is found to be in a sister group position to all other Branchiura, which in this analysis include six Argulus and one Chonopeltis sequences. Based on the molecular phylogeny a likely evolutionary scenario is that the ancestral branchiuran used hooks (on the first maxilla) for attachment, as seen in Dolops, of which the proximal part was subsequently modified into suction discs in Argulus and Chonopeltis (and Dipteropeltis). The sister group relationship of the Branchiura and Pentastomida is confirmed based on the most comprehensive taxon sampling until now. No evidence was found for a branchiuran in-group position of the Pentastomida. PMID- 18394960 TI - Mandibular glands of male European beewolves, Philanthus triangulum (Hymenoptera, Crabronidae). AB - Males of a solitary digger wasp, the European beewolf, Philanthus triangulum, possess large mandibular glands that have been reported to produce a scent marking pheromone. We analysed the morphology and ultrastructure of these glands using light microscopy as well as scanning and transmission electron microscopy. The paired glands are located laterally in the head and each side consists of a larger and a smaller part. Both parts possess a collecting duct each with distinct openings at the mandible base. However, the collecting duct of the larger part is additionally connected to the pharynx through a lateral extension. The collecting ducts are bordered by a monolayered epithelium lined with cuticle that exhibits conspicuous ramified protuberances. About 1400 acini consisting of class 3 gland cells surround the ducts and are connected to them through conducting canals. The main components in the cytoplasm of these gland cells are mitochondria, well-developed smooth endoplasmatic reticulum, and electron lucent vesicles suggesting a high secretory activity. The connection between the large gland parts and the pharynx suggests that the secretion of the mandibular glands might not only be delivered directly onto the mandibles but might also be transported to and stored in the postpharyngeal gland. PMID- 18394961 TI - Inhibition of PI-3 kinase for treating respiratory disease: good idea or bad idea? AB - Inhibition of one or more members of the phosphoinositide 3-kinase (PI3K) family for the treatment of respiratory diseases remains the goal of many pharmaceutical companies over the past 20 years. Here we briefly review the PI3K family, then focus on the assessment of each isoform as a drug discovery target. The rationale for PI3Kalpha inhibition in the treatment of lung cancer, and PI3Kbeta inhibitors in pulmonary thrombotic processes, are balanced with a potential side effect profile affecting metabolism and/or foetal development. Roles for PI3Kdelta in inflammatory lung diseases and PI3Kgamma in asthma are weighed against the consequences of manipulating key immune cell populations. We also discuss the current status and future potential of PI3K inhibitors in respiratory disease. PMID- 18394962 TI - Innate immune 'self' recognition: a role for CD47-SIRPalpha interactions in hematopoietic stem cell transplantation. AB - Self-nonself discrimination is a central property of the immune system. This paradigm was originally established in the context of tissue transplantation, leading to the discovery of major histocompatibility complex molecules as signals of 'self'. However, accumulating evidence has shown that innate immune cells are regulated in a similar fashion. Recent evidence has suggested that interactions between the 'self' molecule CD47 and the innate inhibitory receptor signal regulatory protein-alpha expressed on macrophages may be a critical determinant of transplant engraftment, supporting the concept that 'self'-awareness is a general property of all immune cells. PMID- 18394963 TI - Tissue distribution of lymphocytes and plasma cells and the role of the gut. PMID- 18394965 TI - The specificity and sensitivity of transcranial ultrasound in the differential diagnosis of Parkinson's disease: a prospective blinded study. AB - BACKGROUND: Increased echogenicity of the substantia nigra (SN), as determined by transcranial sonography (TCS), is characteristic of idiopathic Parkinson's disease (iPD). The results of initial retrospective studies indicate that this ultrasound sign is specific for iPD and can help to differentiate it from atypical parkinsonian syndromes (aPS); however, these early studies were done in patients with later disease stages and known clinical diagnosis. We aimed to determine the diagnostic value of TCS in the early stages of parkinsonian syndromes, when the clinical symptoms often do not enable a definite diagnosis to be made. METHODS: 60 patients who presented with the first, but still unclear, clinical symptoms of parkinsonism had TCS in this prospective blinded study. Investigators were blinded to the results of the clinical investigations, the ultrasound findings, and the diagnosis at time of investigation. The patients were followed-up every 3 months for 1 year to assess and re-evaluate the clinical symptoms. The patients in whom a clinical diagnosis could not be made with certainty were investigated with raclopride PET or dopamine transporter single photon emission computed tomography (SPECT), or both. FINDINGS: A clinical diagnosis of parkinsonism could not be established at baseline in 38 patients. At 12 months, 39 patients were clinically categorised as having iPD. Compared with endpoint diagnosis, the sensitivity of TCS at baseline was 90%7% and the specificity was 82.4%; the positive predictive value of TCS for iPD was 92.9% and the classification accuracy was 88.3%. INTERPRETATION: TCS is an easy to implement, non-invasive, and inexpensive technique that could help in the early differential diagnosis of parkinsonian syndromes. The routine use of TCS in the clinic could enable disease-specific therapy to be started earlier. FUNDING: Michael J Fox Foundation for Parkinson's Research. PMID- 18394966 TI - Transcranial ultrasound in Parkinson's disease. PMID- 18394967 TI - Association of the interleukin (IL)-16 gene polymorphisms with Graves' disease. AB - Interleukin (IL)-16 was one of the cytokines with the function of T helper cell recruitment, whose expression in the thyrocyte and orbital fibroblast of Graves' disease (GD) patients was increased. Recently association of IL-16 gene polymorphisms with autoimmune diseases had been reported. However, there was little known about the impact of IL-16 gene polymorphisms on GD. In this study, we performed a case-control association study of three tagSNPs (rs4778889 rs1131445-rs4778641) within the IL-16 gene on 258 patients with GD and 208 healthy subjects in the Chinese population. Our data showed that common IL-16 variants were associated with GD (P=0.013-0.0186) and Graves' disease associated ophthalmopathy (GO) (P=0.0033-0.041). A novel protective haplotype containing the three tagSNPs (C-T-C) was observed in association with GO (P=0.013). In conclusion, IL-16 gene was significantly associated with susceptibility to Graves' disease and Graves' disease associated ophthalmopathy in the Chinese population. PMID- 18394969 TI - From user needs to system specifications: multi-disciplinary thematic seminars as a collaborative design method for development of health information systems. AB - This paper presents a new multi-disciplinary method for user needs analysis and requirements specification in the context of health information systems based on established theories from the fields of participatory design and computer supported cooperative work (CSCW). Whereas conventional methods imply a separate, sequential needs analysis for each profession, the "multi-disciplinary thematic seminar" (MdTS) method uses a collaborative design process. Application of the method in elderly homecare resulted in prototypes that were well adapted to the intended user groups. Vital information in the points of intersection between different care professions was elicited and a holistic view of the entire care process was obtained. Health informatics-usability specialists and clinical domain experts are necessary to apply the method. Although user needs acquisition can be time-consuming, MdTS was perceived to efficiently identify in-context user needs, and transformed these directly into requirements specifications. Consequently the method was perceived to expedite the entire ICT implementation process. PMID- 18394968 TI - Local and systemic release of cytokines, nerve growth factor, prostaglandin E2, and substance P in incisional wounds and serum following cesarean delivery. AB - The objectives of this study were to test the feasibility of measuring inflammatory and nociceptive biochemical mediators at the surgical site and to evaluate the relationship between wound and serum levels as well as determine any associations between mediator release, pain, and analgesic consumption after cesarean delivery. Twenty healthy women undergoing elective cesarean delivery with spinal anesthesia were enrolled. Wound exudate and serum mediators, pain scores, and analgesic consumption were measured at 1, 6, 24, and 48 hours after cesarean. In wound exudate, 19 of 20 mediators were reliably detected including interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, tumor necrosis factor-alpha, interferon-gamma, granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein 1 (MCP-1) and macrophage inflammatory protein 1 (MIP-1beta), nerve growth factor (NGF), prostaglandin E2 (PG-E2), and substance P. Wound PG-E2 and various cytokines peaked early, whereas NGF showed a more delayed release. There were no correlations between the concentration versus time profile of wound and serum cytokines. Analgesic consumption during the first 24 hours after surgery was negatively correlated with IL-1beta, IL-6, and G-CSF in the wound exudate. This study demonstrates the feasibility of collecting and measuring nociceptive and inflammatory mediators in surgical wounds at specific time points. The lack of significant correlations between wound and serum levels emphasizes the importance of determining site-specific release if localized pathologies are to be studied. PERSPECTIVE: This study demonstrates the feasibility of measuring real-time nociceptive and inflammatory mediators in surgical wounds. Our findings confirm the lack of correlation between wound and serum levels of many pro-inflammatory and anti-inflammatory cytokines and nerve growth factor. PMID- 18394970 TI - Positive selection in penicillin-binding proteins 1a, 2b, and 2x from Streptococcus pneumoniae and its correlation with amoxicillin resistance development. AB - The efficacy of beta-lactam antibiotics in Streptococcus pneumoniae has been compromised because of the development of altered penicillin-binding proteins (PBPs), however, this has been less so for amoxicillin than for penicillin. Recently, there have been a number of important methods developed to detect molecular adaptation in protein coding genes. The purpose of this study is to employ modern molecular selection approaches to predict sites under positive selection pressure in PBPs, derived from a large international S. pneumoniae collection of amoxicillin resistant and susceptible isolates, and encompassing a comparative data set of 354 pbp1a, 335 pbp2b, and 389 pbp2x gene sequences. A correspondence discriminant analysis (CDA) of positively selected pbp sites and amoxicillin MIC (minimum inhibitory concentration) values is then used to detect sites under positive selection pressure that are important in discriminating different amoxicillin MICs. Molecular adaptation was evident throughout PBP2X, with numerous positively selected sites in both the transpeptidase (TP) and C terminal domains, strongly correlated with discriminating amoxicillin MICs. In the case of PBP1A positive selection was present in the glycosyltransfer (GT), TP and C-terminal domains. Sites within the TP domain tended to be correlated with the discrimination of low from intermediate MICs, whereas sites within the C terminal tail, with a discrimination of intermediate from fully resistant. Most of the positively selected sites within PBP2B were in the N-terminal domain and were not correlated with amoxicillin MICs, however, several sites taken from the literature for the TP domain were strongly associated with discriminating high from intermediate level amoxicillin resistance. Many of the positively selected sites could be directly associated with functional inferences based on the crystal structures of these proteins. Our results suggest that clinical emphasis on TP domain sequences of these proteins may result in missing information relevant to antibiotic resistance development. PMID- 18394971 TI - Host genetics in African trypanosomiasis. AB - In Africa, the protozoan parasite of the genus Trypanosoma causes animal (AAT) and human African trypanosomiasis (HAT). These diseases are responsible for considerable mortality and economic losses, and until now the drugs commonly used have often been very toxic and expensive, with no vaccine available. A range of clinical presentations, from chronic to acute symptoms, is observed in both AAT and HAT. Host, parasite, and environmental factors are likely to be involved in this clinical variability. In AAT, some West African cattle (N'Dama, Bos taurus) have the ability to better control the disease development (and therefore to remain productive) than other taurine breeds (Zebu, Bos indicus). This phenomenon is called trypanotolerance and seems to have major genetic components. In humans, tolerance/resistance to the disease is suspected, however, this needs confirmation. This review focuses on recent advances made in the field of host genetics in African trypanosomiasis in animals (mouse and bovine) and humans. The perspectives for the development of new control strategies and their applications as well as a better understanding of the physiopathology of the disease are discussed. PMID- 18394972 TI - Prevalence and diversity of Lyme borreliosis bacteria in marine birds. AB - A potential role of seabirds in spreading Lyme disease (LB) spirochetes over large spatial scales was suggested more than 10 years ago when Borrelia garinii was observed in marine birds of both hemispheres. Since then, there have been few studies examining the diversity of Borrelia spp. circulating in seabirds, or the potential interaction between terrestrial and marine disease cycles. To explore these aspects, we tested 402 Ixodes uriae ticks collected from five colonial seabird species by amplification of the flaB gene. Both the average prevalence (26.0%+/-3.9) and diversity of LB spirochetes was high. Phylogenetic analyses grouped marine isolates in two main clades: one associated with B. garinii and another with B. lusitaniae, a genospecies typically associated with lizards. One sequence also clustered most closely with B. burgdorferi sensu stricto. Prevalence in ticks varied both among seabird species within colonies and among colonies. However, there was no clear association between different Borrelia isolates and a given seabird host species. Our findings indicate that LB spirochetes circulating in the marine system are more diverse than previously described and support the hypothesis that seabirds may be an important component in the global epidemiology and evolution of Lyme disease. Future work should help determine the extent to which isolates are shared between marine and terrestrial systems. PMID- 18394973 TI - Ageing and urban-rural drift in Nigeria: coping or dispensing with city accommodation in retirement. AB - The massive press attentions attracted by the plight of pensioners in Nigeria appear to provide the extrinsic nudge and popular support for the 2004 pension reform. In some sense, these sets of exposes offered a new peephole into the yawning care and support gap for the elderly in the country. Against the background of counteracting processes of population ageing and receding formal and informal social protection and support mechanisms, the country appears ill prepared to tackle the unfolding old age bulge and its myriad challenges. The paper addresses one of the most basic needs and choices confronting older people after retirement: the resolve to cope or dispense with city (or urban) accommodation. Such decisions have become increasingly critical not only because they are principal determinants of urban-rural drift but also because of the changing circumstances (of weakening support systems and increased vulnerability) under which they are made. The paper explores these important ageing issue and their implications for policy in Nigeria. PMID- 18394975 TI - Electrical percutaneous tibial stimulation modulates within-a-breath respiratory drive in man. AB - A sharp intake of breath followed by a strong vocalisation is widely observed in response to acute pain although its function and mechanism is poorly understood. This study investigated the effect of percutaneous (overlying the tibial bone) electrical stimulation delivered early (20-30% of inspiratory time) during inspiration (INSP) or expiration (EXP) (20-30% of expiratory time) at sensory intensities at (100%), above (125%) and below (50% and 75%) the pre-determined pain threshold (PT), upon within-a-breath respiratory parameters (via pneumotachography). All INSP stimulation intensities provoked significant inspiratory time shortening thereby elevating mean inspiratory flow. Tidal volume, but not peak flow was increased in response to 100% PT and 125% PT stimulation (vs. PRE). Shortening and increased tidal volume combined to evoke significant mean inspiratory airflow increments. In contrast, EXP stimulation failed to evoke any effect. Thus, our study provides evidence of a within-a breath inspiratory-specific, augmentory response to noxious stimulation. PMID- 18394974 TI - Opioidergic and dopaminergic modulation of respiration. AB - Opioids, dopamine and their receptors are present in many regions of the bulbar respiratory network. The physiological importance of endogenous opioids to respiratory control has not been explicitly demonstrated. Nonetheless, studies of opioidergic respiratory mechanisms are important because synthetic opiate drugs have respiratory side effects that in some situations pose health risks and limit their therapeutic usefulness. They can depress breathing depth and rate, blunt respiratory responsiveness to CO2 and hypoxia, increase upper airway resistance and reduce pulmonary compliance. The opiate respiratory disturbances are mainly due to agonist activation of mu- and delta-subtypes of receptor and involve specific types of respiratory-related neurons in the ventrolateral medulla and the dorsolateral pons. Endogenous dopaminergic modulation in the CNS and carotid bodies enhances CO2-dependent respiratory drive and depresses hypoxic drive. In the CNS, synthetic agonists with selectivity for D1-and D4-types of receptor slow respiratory rhythm, whereas D2-selective agonists modulate acute and chronic responses to hypoxia. D1-receptor agonists also act centrally to increase respiratory responsiveness to CO2, and counteract opiate blunting of CO2 dependent respiratory drive and depression of breathing. Cellular targets and intracellular mechanisms responsible for opioidergic and dopaminergic respiratory effects for the most part remain to be determined. PMID- 18394976 TI - Correlation between predicted and actual consequences of capsule endoscopy on patient management. AB - BACKGROUND: Capsule endoscopy (CE) is a relatively new diagnostic modality in the evaluation of patients with suspected small bowel pathology. It is unclear to what extent physicians are able to predict the clinical consequences of CE on patient management. METHODS: In this prospective study, 180 consecutive CE examinations were analysed. Prior to CE, referring physicians were asked to indicate the consequences of CE according to potential different CE outcomes. The influence of CE on patient management was determined with at least 1 year follow up. Management consequences were defined as major (surgical or endoscopic intervention, or medical therapy) or minor (nonspecific therapy, including iron supplementation, or no further diagnostic tests). RESULTS: CE led to major management consequences in 32% of cases. Of patients with obscure gastrointestinal bleeding and normal CE findings, 91% were independent of blood transfusions and experienced no further bleeding episodes during a mean follow-up of 33 months. In 78% of 118 cases that were evaluated, the actual consequences of CE matched the consequences predicted by the referring physicians. CONCLUSION: CE had a major impact on patient management in about one third of investigations. In the majority of cases, physicians adequately predicted the clinical consequences of CE. PMID- 18394977 TI - Nutcracker oesophagus: association with chest pain and dysphagia controlling for gastro-oesophageal reflux. AB - BACKGROUND: The association between nutcracker oesophagus, gastro-oesophageal reflux and their symptoms is controversial. AIM: To evaluate the association of nutcracker oesophagus with chest pain and dysphagia controlling for gastro oesophageal reflux. METHODS: From a database of 935 consecutive patients investigated with oesophageal manometry and pH-metry, we selected all patients with nutcracker oesophagus including diffuse and segmental patterns. Patients with normal oesophageal peristalsis served as controls. Symptoms assessment, manometry testing and 24h oesophageal pH monitoring off acid-suppressive medications were performed following a standardized protocol. The associations between nutcracker oesophagus and symptoms were assessed by logistic regression analysis. RESULTS: Nutcracker oesophagus was found in 60 patients (6.4%), of which 30 had diffuse nutcracker oesophagus and 30 had segmental nutcracker oesophagus. The control group was composed by 656 patients with normal oesophageal peristalsis. Diffuse nutcracker oesophagus was associated with chest pain (odds ratio 4.3; 95% CI 1.9-9.9; P<0.0001) and dysphagia (odds ratio 5.3; 95% CI 2.3-12.2; P<0.0001), whereas segmental nutcracker oesophagus was associated with chest pain (odds ratio 2.8; 95% CI 1.1-6.9; P=0.026), controlling for total oesophageal acid exposure, age, sex and lower oesophageal sphincter (LOS) pressure. CONCLUSION: This study suggests that both diffuse and segmental nutcracker oesophagus should be regarded as meaningful abnormalities and not mere manometric curiosities. PMID- 18394978 TI - Feasibility of electrocautery snaring as the final step of endoscopic submucosal dissection for stomach epithelial neoplasms. AB - BACKGROUND: Endoscopic submucosal dissection (ESD) is a novel endoluminal technique that permits the resection of gastric neoplasms. AIM: To analyse the feasibility of snaring as the final step of ESD. PATIENTS AND METHODS: One hundred and ninety-nine consecutive gastric neoplasms resected by four ESD experts from January 2004 to May 2007 were investigated. Forty-five (22.6%) were finally resected finally using a snare. Rates of en bloc resection, complete (R0 plus en bloc) resection, mean operation time, and complications were assessed between the snaring and the non-snaring groups. RESULTS: En bloc resection rate was significantly lower and delayed bleeding rate was significantly higher in the snaring group than in the non-snaring group (91.1% [41/45] vs. 100% [154/154], 11.1% [5/45] vs. 1.9% [3/154], respectively), although complete resection rate (86.7% [39/45] vs. 92.9% [143/154]) and mean operation time (70.2 min vs. 75.8 min) were not significantly different between the two groups. Six perforation cases (3 [6.7%] in the snaring group, 3 [1.9%] in the non-snaring group) were observed, but snaring did not lead to perforation in any case. When the subjects were divided into small (< or = 2 cm) and large (> 2 cm) tumours, en bloc resection rate in large tumours was still significantly different between the groups (76.9% [10/13] vs. 100% [67/67]), whereas in small tumours it was no longer significantly different (96.9% [31/32] vs. 100% [87/87]). CONCLUSIONS: Snaring may facilitate successful ESD for smaller tumours, but multiple-piece resection should be taken into account especially for larger tumours. PMID- 18394979 TI - Single nucleotide polymorphisms in human Paneth cell defensin A5 may confer susceptibility to inflammatory bowel disease in a New Zealand Caucasian population. AB - BACKGROUND: Human Paneth cell alpha-defensins, especially DEFA5, are involved in maintaining homeostasis of the human microbial microflora. Since breakdown of normal mucosal antibacterial defence occurs in inflammatory bowel disease (IBD), variants in the DEFA5 gene could be associated with IBD risk. SUBJECTS: A cohort of 25 patients with indeterminate colitis (IC), 405 with ulcerative colitis (UC), and 385 with Crohn's disease (CD), were compared with 201 control individuals from the Canterbury region in New Zealand. METHODS: A 15 kb haplotype block surrounding DEFA5 contained 35 HapMap markers which were polymorphic in Caucasians. Four markers (A-D) were selected to tag 27 of the 35 markers at r(2)>0.68, and were genotyped in DNA samples. RESULTS: Minor allele frequencies for all single nucleotide polymorphisms (SNPs) were somewhat elevated in patients. Subgroup analysis showed SNP A had odds ratio 1.44 in UC patients with pancolitis (95% C.I. 1.07-1.94), SNP B odds ratio 2.37 in CD patients with onset prior to 17 years age (95% C.I. 1.12-5.03), SNP C odds ratio 1.68 in UC patients with left colonic localisation (95% C.I. 1.12-2.52), and SNP D had odds ratio 1.56 in CD patients with one or more relatives with IBD (95% C.I. 1.03-2.35). Two two-marker haplotypes and one three-marker haplotype were associated with UC (p values 0.025-0.05). CONCLUSIONS: The SNPs genotyped in our study were surrogates for common variants, and observed associations between these and IBD status are likely due to linkage disequilibrium with a functional common DEFA5 variant. Identifying such functional variants will be prioritized in subsequent work. PMID- 18394980 TI - An in vitro evaluation of the Ca/P ratio for the cytocompatibility of nano-to micron particulate calcium phosphates for bone regeneration. AB - Calcium phosphate based bioceramics have been widely used for orthopedic applications due to their chemical similarity to natural bone. The Ca/P stoichiometry of calcium phosphates strongly influences their performance under biological conditions, which have not yet been fully elucidated to date. For this reason, the objective of this in vitro study was to understand the relationship between the Ca/P ratio of nano-to-micron particulate calcium phosphate substrates and their biological properties, such as osteoblast (bone-forming cell) viability, collagen production, alkaline phosphatase activity and nitric oxide (NO) production. A group of calcium phosphates with Ca/P ratios between 0.5 and 2.5 were obtained by intentionally adjusting the Ca/P stoichiometry of the initial reactants necessary for calcium phosphate precipitation. For samples with 0.5 and 0.75 Ca/P ratios, tricalcium phosphate (TCP) and Ca(2)P(2)O(7) phases were observed. In contrast, for samples with 1.0 and 1.33 Ca/P ratios, the only stable phase was TCP. For samples with a 1.5 Ca/P ratio, the TCP phase was dominant; however, small amounts of the hydroxyapatite (HA) phase started to appear. For samples with a 1.6 Ca/P ratio, the HA phase was dominant. Lastly, for samples with 2.0 and 2.5 Ca/P ratios, the CaO phase started to appear in the HA phase which was the dominant phase. Moreover, the average grain size and the average pore size decreased from micron-scale (e.g. 1370nm for a 0.5 Ca/P ratio) to nano-scale (e.g. 262nm for a 2.5 Ca/P ratio) with increasing Ca/P ratios. The porosity (%) of calcium phosphate substrates also decreased with increasing Ca/P ratios. Previous in vitro results demonstrated increased osteoblast adhesion on calcium phosphates with higher Ca/P ratios (up to 2.5). The present study showed that the collagen production by osteoblasts was similar between all the calcium phosphates but slightly lower with a 1.6 Ca/P ratio. Greater alkaline phosphatase activity by osteoblasts was observed in all the cultures with various calcium phosphates (0.5-2.5 Ca/P ratios) than in the control (only cells in culture). Ca/P ratios of <2 and 1 optimized osteoblast viability and promoted alkaline phosphatase activity in osteoblasts, respectively. However, the presence of the CaO phase in Ca/P ratios 2.0 increased osteoblast NO production and decreased osteoblast viability. In summary, this study provided evidence that the Ca/P ratio of calcium phosphate is a very important factor that should be considered when selecting nano-to-micron particulate calcium phosphates for various orthopedic applications. PMID- 18394981 TI - Identification of the functional activity of the [A-4] amelogenin gene splice product in newborn mouse ameloblasts. AB - In the mouse tooth organ, shortly after birth, ameloblasts acquire their secretory phenotype, which is characterized by the prominent expression and subsequent secretion of two isoforms of amelogenin, M180 and M59 (LRAP, [A-4]). Amelogenin deposition into the ameloblast extracellular matrix promotes enamel biomineralization. A complex set of intercellular signaling events, reciprocal communications between the developing oral epithelium and its underlying dental mesenchyme, guide the expression of amelogenin mRNA, and limit it to a defined period of tooth development. In tooth germ organ culture, addition of the [A-4] isoform, lacking amelogenin exon 4 and exon 6 segments a, b, c, was shown to affect ameloblast development. To understand the basis for this regulatory activity, we have studied the effects of r[A-4] on ameloblast-like LS8 cells, and the role of the putative [A-4] cell surface receptor, LAMP1, as well as the related receptor LAMP3. In the LS8 cells, the expression of the spliced isoforms of amelogenin, LAMP1, and LAMP3 were identified by RT-PCR, and real-time PCR semi quantitative analysis assessed the modulation of M180 message. M180 mRNA was up regulated by exogenous [A-4], and this was further increased by blockade of LAMP1, suggesting additive effects between the intracellular signaling pathways activated by the discrete agonists. Immunofluorescence staining identified the patterns of [A-4] and LAMP1 localization in LS8 cells. Internalized r[A-4] was co localized with LAMP1 in late endosomal/lysosomal compartments. Thus, the LAMP1 and [A-4] intracellular sorting pathways are interrelated. The nitric oxide (NO) signaling pathway was activated by exogenous [A-4]. [A-4] modulated inducible nitric oxide synthase (iNOS, NOS2) and endothelial nitric oxide synthase (eNOS, NOS3) expression, albeit, to different extents. NOS2 was significantly up regulated after 4 h, while NOS3 increased slightly after 24 h. Co-treatment of LS8 cells with r[A-4] and anti-LAMP1 antibodies further enhanced NOS2 expression. Anti-LAMP1 antibodies did not abrogate NO production in LS8 cells treated for 4 h with r[A-4], but the iNOS inhibitor, l-Nil, down-regulated both NO production and the expression of M180 mRNA. These data suggest that [A-4] modulates M180 mRNA expression, partly, via the NO signaling pathway. PMID- 18394982 TI - The immortal strand hypothesis: how could it work? PMID- 18394983 TI - Sorting out small RNAs. AB - Small RNAs carry out their functions by guiding Argonaute (AGO) proteins to their targets. Diverse types of small RNAs and multiple AGO proteins exist in most eukaryotic species, but how small RNAs are sorted into specific AGO complexes remains unclear. Two papers in this issue (Mi et al., 2008; Montgomery et al., 2008) now reveal the importance of the 5' terminal nucleotide of the small RNA in the sorting process in Arabidopsis. PMID- 18394984 TI - TGFbeta primes breast tumor cells for metastasis. AB - The microenvironment of a tumor is known to influence tumor progression and spread to distant sites (metastasis). Padua et al. (2008) now show that transient exposure of breast cancer cells to the signaling molecule transforming growth factor beta (TGFbeta) promotes their extravasation from blood vessels and entry into the lung by upregulation of the adipokine angiopoietin-like 4 (ANGPTL4). Their work shows that the later stages of metastasis can be influenced by transient signals produced in the primary tumor microenvironment. PMID- 18394985 TI - A tRNA with oncogenic capacity. AB - Overexpression of Brf1, a transcription factor of the RNA polymerase III apparatus, can transform cells in vitro and cause tumor formation in vivo. Marshall et al. (2008) now show that one of the transcriptional products of RNA polymerase III, the initiator tRNA(Met), mediates this effect, revealing an unexpected role for this tRNA in tumorigenesis. PMID- 18394986 TI - A new gene for auxin synthesis. AB - There is much interest in understanding the pathways that trigger biosynthesis of the plant hormone auxin. In this issue, Stepanova et al. (2008) and Tao et al. (2008) reveal that a small family of tryptophan aminotransferases catalyze formation of indole-3-pyruvic acid (IPA) from L-tryptophan (L-Trp), the first step in a pathway for auxin biosynthesis. PMID- 18394987 TI - A SNO storm in skeletal muscle. AB - Dysregulated S-nitrosylation of proteins characterizes a broad array of human disorders, but its role in disease etiology is not well understood. Two new studies (Durham et al., 2008; Bellinger et al., 2008) now show that hyper-S nitrosylation of the ryanodine receptor calcium release channel (RyR1) in skeletal muscle disrupts calcium ion flux. This disruption underlies the impaired contractility and cellular damage of skeletal muscle during strenuous exercise and in a spectrum of congenital muscle disorders including malignant hyperthermia. PMID- 18394988 TI - Eph-ephrin bidirectional signaling in physiology and disease. AB - Receptor tyrosine kinases of the Eph family bind to cell surface-associated ephrin ligands on neighboring cells. The ensuing bidirectional signals have emerged as a major form of contact-dependent communication between cells. New findings reveal that Eph receptors and ephrins coordinate not only developmental processes but also the normal physiology and homeostasis of many adult organs. Imbalance of Eph/ephrin function may therefore contribute to a variety of diseases. The challenge now is to better understand the complex and seemingly paradoxical signaling mechanisms of Eph receptors and ephrins, which will enable effective strategies to target these proteins in the treatment of diseases such as diabetes and cancer. PMID- 18394989 TI - RyR1 S-nitrosylation underlies environmental heat stroke and sudden death in Y522S RyR1 knockin mice. AB - Mice with a malignant hyperthermia mutation (Y522S) in the ryanodine receptor (RyR1) display muscle contractures, rhabdomyolysis, and death in response to elevated environmental temperatures. We demonstrate that this mutation in RyR1 causes Ca(2+) leak, which drives increased generation of reactive nitrogen species (RNS). Subsequent S-nitrosylation of the mutant RyR1 increases its temperature sensitivity for activation, producing muscle contractures upon exposure to elevated temperatures. The Y522S mutation in humans is associated with central core disease. Many mitochondria in the muscle of heterozygous Y522S mice are swollen and misshapen. The mutant muscle displays decreased force production and increased mitochondrial lipid peroxidation with aging. Chronic treatment with N-acetylcysteine protects against mitochondrial oxidative damage and the decline in force generation. We propose a feed-forward cyclic mechanism that increases the temperature sensitivity of RyR1 activation and underlies heat stroke and sudden death. The cycle eventually produces a myopathy with damaged mitochondria. PMID- 18394990 TI - TGFbeta primes breast tumors for lung metastasis seeding through angiopoietin like 4. AB - Cells released from primary tumors seed metastases to specific organs by a nonrandom process, implying the involvement of biologically selective mechanisms. Based on clinical, functional, and molecular evidence, we show that the cytokine TGFbeta in the breast tumor microenvironment primes cancer cells for metastasis to the lungs. Central to this process is the induction of angiopoietin-like 4 (ANGPTL4) by TGFbeta via the Smad signaling pathway. TGFbeta induction of Angptl4 in cancer cells that are about to enter the circulation enhances their subsequent retention in the lungs, but not in the bone. Tumor cell-derived Angptl4 disrupts vascular endothelial cell-cell junctions, increases the permeability of lung capillaries, and facilitates the trans-endothelial passage of tumor cells. These results suggest a mechanism for metastasis whereby a cytokine in the primary tumor microenvironment induces the expression of another cytokine in departing tumor cells, empowering these cells to disrupt lung capillary walls and seed pulmonary metastases. PMID- 18394991 TI - Elevated tRNA(iMet) synthesis can drive cell proliferation and oncogenic transformation. AB - Characteristics of transformed and tumor cells include increased levels of protein synthesis and elevated expression of RNA polymerase (pol) III products, such as tRNAs and 5S rRNA. However, whether deregulated pol III transcription contributes to transformation has been unclear. Generating cell lines expressing an inducible pol III-specific transcription factor, Brf1, allowed us to raise tRNA and 5S rRNA levels specifically. Brf1 induction caused an increase in cell proliferation and oncogenic transformation, whereas depletion of Brf1 impeded transformation. Among the gene products induced by Brf1 is the tRNA(iMet) that initiates polypeptide synthesis. Overexpression of tRNA(iMet) is sufficient to stimulate cell proliferation and allow immortalized fibroblasts to form foci in culture and tumors in mice. The data indicate that elevated tRNA synthesis can promote cellular transformation. PMID- 18394992 TI - Independent positioning and action of Escherichia coli replisomes in live cells. AB - A prevalent view of DNA replication has been that it is carried out in fixed "replication factories." By tracking the progression of sister replication forks with respect to genetic loci in live Escherichia coli, we show that at initiation replisomes assemble at replication origins irrespective of where the origins are positioned within the cell. Sister replisomes separate and move to opposite cell halves shortly after initiation, migrating outwards as replication proceeds and both returning to midcell as replication termination approaches. DNA polymerase is maintained at stalled replication forks, and over short intervals of time replisomes are more dynamic than genetic loci. The data are inconsistent with models in which replisomes associated with sister forks act within a fixed replication factory. We conclude that independent replication forks follow the path of the compacted chromosomal DNA, with no structure other than DNA anchoring the replisome to any particular cellular region. PMID- 18394993 TI - Resolution of sister centromeres requires RanBP2-mediated SUMOylation of topoisomerase IIalpha. AB - RanBP2 is a nucleoporin with SUMO E3 ligase activity that functions in both nucleocytoplasmic transport and mitosis. However, the biological relevance of RanBP2 and the in vivo targets of its E3 ligase activity are unknown. Here we show that animals with low amounts of RanBP2 develop severe aneuploidy in the absence of overt transport defects. The main chromosome segregation defect in cells from these mice is anaphase-bridge formation. Topoisomerase IIalpha (Topo IIalpha), which decatenates sister centromeres prior to anaphase onset to prevent bridges, fails to accumulate at inner centromeres when RanBP2 levels are low. We find that RanBP2 sumoylates Topo IIalpha in mitosis and that this modification is required for its proper localization to inner centromeres. Furthermore, mice with low amounts of RanBP2 are highly sensitive to tumor formation. Together, these data identify RanBP2 as a chromosomal instability gene that regulates Topo IIalpha by sumoylation and suppresses tumorigenesis. PMID- 18394994 TI - Monitoring protein conformation along the pathway of chaperonin-assisted folding. AB - The GroEL/GroES chaperonin system mediates protein folding in the bacterial cytosol. Newly synthesized proteins reach GroEL via transfer from upstream chaperones such as DnaK/DnaJ (Hsp70). Here we employed single molecule and ensemble FRET to monitor the conformational transitions of a model substrate as it proceeds along this chaperone pathway. We find that DnaK/DnaJ stabilizes the protein in collapsed states that fold exceedingly slowly. Transfer to GroEL results in unfolding, with a fraction of molecules reaching locally highly expanded conformations. ATP-induced domain movements in GroEL cause transient further unfolding and rapid mobilization of protein segments with moderate hydrophobicity, allowing partial compaction on the GroEL surface. The more hydrophobic regions are released upon subsequent protein encapsulation in the central GroEL cavity by GroES, completing compaction and allowing rapid folding. Segmental chain release and compaction may be important in avoiding misfolding by proteins that fail to fold efficiently through spontaneous hydrophobic collapse. PMID- 18394995 TI - Structural mechanism of demethylation and inactivation of protein phosphatase 2A. AB - Protein phosphatase 2A (PP2A) is an important serine/threonine phosphatase that plays a role in many biological processes. Reversible carboxyl methylation of the PP2A catalytic subunit is an essential regulatory mechanism for its function. Demethylation and negative regulation of PP2A is mediated by a PP2A-specific methylesterase PME-1, which is conserved from yeast to humans. However, the underlying mechanism of PME-1 function remains enigmatic. Here we report the crystal structures of PME-1 by itself and in complex with a PP2A heterodimeric core enzyme. The structures reveal that PME-1 directly binds to the active site of PP2A and that this interaction results in the activation of PME-1 by rearranging the catalytic triad into an active conformation. Strikingly, these interactions also lead to inactivation of PP2A by evicting the manganese ions that are required for the phosphatase activity of PP2A. These observations identify a dual role of PME-1 that regulates PP2A activation, methylation, and holoenzyme assembly in cells. PMID- 18394996 TI - Rapid synthesis of auxin via a new tryptophan-dependent pathway is required for shade avoidance in plants. AB - Plants grown at high densities perceive a decrease in the red to far-red (R:FR) ratio of incoming light, resulting from absorption of red light by canopy leaves and reflection of far-red light from neighboring plants. These changes in light quality trigger a series of responses known collectively as the shade avoidance syndrome. During shade avoidance, stems elongate at the expense of leaf and storage organ expansion, branching is inhibited, and flowering is accelerated. We identified several loci in Arabidopsis, mutations in which lead to plants defective in multiple shade avoidance responses. Here we describe TAA1, an aminotransferase, and show that TAA1 catalyzes the formation of indole-3-pyruvic acid (IPA) from L-tryptophan (L-Trp), the first step in a previously proposed, but uncharacterized, auxin biosynthetic pathway. This pathway is rapidly deployed to synthesize auxin at the high levels required to initiate the multiple changes in body plan associated with shade avoidance. PMID- 18394997 TI - TAA1-mediated auxin biosynthesis is essential for hormone crosstalk and plant development. AB - Plants have evolved a tremendous ability to respond to environmental changes by adapting their growth and development. The interaction between hormonal and developmental signals is a critical mechanism in the generation of this enormous plasticity. A good example is the response to the hormone ethylene that depends on tissue type, developmental stage, and environmental conditions. By characterizing the Arabidopsis wei8 mutant, we have found that a small family of genes mediates tissue-specific responses to ethylene. Biochemical studies revealed that WEI8 encodes a long-anticipated tryptophan aminotransferase, TAA1, in the essential, yet genetically uncharacterized, indole-3-pyruvic acid (IPA) branch of the auxin biosynthetic pathway. Analysis of TAA1 and its paralogues revealed a link between local auxin production, tissue-specific ethylene effects, and organ development. Thus, the IPA route of auxin production is key to generating robust auxin gradients in response to environmental and developmental cues. PMID- 18394998 TI - SnapShot: The splicing regulatory machinery. PMID- 18394999 TI - A mobility shift detection method for DNA methylation analysis using phosphate affinity polyacrylamide gel electrophoresis. AB - We describe a procedure for DNA methylation analysis using the bisulfite-mediated cytosine-to-uracil conversion of a target DNA followed by methylation-specific polymerase chain reaction (MSP) and phosphate affinity polyacrylamide gel electrophoresis (PAGE). The MSP was performed using a 1:1 mixture of 5' phosphorylated methylation-specific and 5'-OH non-methylation-specific primers. The PAGE using an immobilized phosphate-binding tag molecule (i.e., a polyacrylamide-bound dizinc(II) complex, Zn(2+)-Phos-tag), which selectively captures the 5'-phosphorylated DNA fragment, enabled the mobility shift detection of the methylation-specific product as a slower migration band. Using this novel procedure, we demonstrated the detection of a methylated cytosine base in a pUC19 plasmid. PMID- 18395000 TI - Measurement of the second osmotic virial coefficient for protein solutions exhibiting monomer-dimer equilibrium. AB - The second osmotic virial coefficient (B) is a measure of solution nonideality that is useful for predicting conditions favorable for protein crystallization and for inhibition of aggregation. Static light scattering is the technique most commonly used to determine B values, typically using protein concentrations less than 5 mg/mL. During static light scattering experiments at low protein concentrations, frequently the protein is assumed to exist either as a single nonassociating species or as a combination of assembly states independent of protein concentration. In the work described here, we examined the limit for ignoring weak reversible dimerization (Kd > or =1 mM) by comparing B values calculated with and without accounting for self-association. Light scattering effects for equilibrium dimer systems with Kd <20 mM and Kd <1 mM will significantly affect apparent B values measured for 20 and 150-kDa proteins, respectively. To interpret correctly light scattering data for monomer-dimer equilibrium systems, we use an expanded coefficient model to account for separate monomer-monomer (B(22)), monomer-dimer (B(23)), and dimer-dimer (B(33)) interactions. PMID- 18395001 TI - Probing the functional tolerance of the b subunit of Escherichia coli ATP synthase for sequence manipulation through a chimera approach. AB - A dimer of 156-residue b subunits forms the peripheral stator stalk of eubacterial ATP synthase. Dimerization is mediated by a sequence with an unusual 11-residue (hendecad) repeat pattern, implying a right-handed coiled coil structure. We investigated the potential for producing functional chimeras in the b subunit of Escherichia coli ATP synthase by replacing parts of its sequence with corresponding regions of the b subunits from other eubacteria, sequences from other polypeptides having similar hendecad patterns, and sequences forming left-handed coiled coils. Replacement of positions 55-110 with corresponding sequences from Bacillus subtilis and Thermotoga maritima b subunits resulted in fully functional chimeras, judged by support of growth on nonfermentable carbon sources. Extension of the T. maritima sequence N-terminally to position 37 or C terminally to position 124 resulted in slower but significant growth, indicating retention of some capacity for oxidative phosphorylation. Portions of the dimerization domain between 55 and 95 could be functionally replaced by segments from two other proteins having a hendecad pattern, the distantly related E subunit of the Chlamydia pneumoniae V-type ATPase and the unrelated Ag84 protein of Mycobacterium tuberculosis. Extension of such sequences to position 110 resulted in loss of function. None of the chimeras that incorporated the leucine zipper of yeast GCN4, or other left-handed coiled coils, supported oxidative phosphorylation, but substantial ATP-dependent proton pumping was observed in membrane vesicles prepared from cells expressing such chimeras. Characterization of chimeric soluble b polypeptides in vitro showed their retention of a predominantly helical structure. The T. maritima b subunit chimera melted cooperatively with a midpoint more than 20 degrees C higher than the normal E. coli sequence. The GCN4 construct melted at a similarly high temperature, but with much reduced cooperativity, suggesting a degree of structural disruption. These studies provide insight into the structural and sequential requirements for stator stalk function. PMID- 18395002 TI - Polycation nanostructured lipid carrier, a novel nonviral vector constructed with triolein for efficient gene delivery. AB - A novel nonviral gene transfer vector was developed by modifying nanostructured lipid carrier (NLC) with cetylated polyethylenimine (PEI). Polycation nanostructured lipid carrier (PNLC) was prepared using the emulsion-solvent evaporation method. Its in vitro gene transfer properties were evaluated in the human lung adenocarcinoma cell line SPC-A1 and Chinese Hamster Ovary (CHO) cells. Enhanced transfection efficiency of PNLC was observed after the addition of triolein to the PNLC formulation and the transfection efficiency of the optimized PNLC was comparable to that of Lipofectamine 2000. In the presence of 10% serum the transfection efficiency of the optimal PNLC was not significantly changed in either cell line, whereas that of Lipofectamine 2000 was greatly decreased in both. Thus, PNLC is an effective nonviral gene transfer vector and the gene delivery activity of PNLC was enhanced after triolein was included into the PNLC formulation. PMID- 18395003 TI - EGF stimulates Cdc42-dependent translocation of SCC antigen to the plasma membrane. AB - Squamous cell carcinoma (SCC) antigen, including intracellular serine protease inhibitors, is widely used as a laboratory marker for cancers of squamous cell origin. Clinical evidences suggest that increased tissue-expression of SCC antigen predicts an invasive phenotype of cancer cells. Herein, we demonstrated that over-expression of SCC antigen increased the rate of EGF-stimulated cell migration. In the search for the underlying molecular mechanism, we have discovered that SCC antigen was translocated to the plasma membrane upon EGF stimulation and co-localized with polymerized-actin at lamellipodia. We further showed that, co-expression of Cdc42, a downstream target of the EGF receptor, enhanced translocation of the SCC antigen, while co-expression of dominant inhibitory Cdc42 diminished its translocation. These results suggest that EGF Cdc42 signal regulates the translocation of SCC antigen to the plasma membrane. Lamellipodia at the leading edge might be a site of action of SCC antigen. PMID- 18395005 TI - JNK activation mediates the apoptosis of xCT-deficient cells. AB - System X(c)(-) is an anionic amino acid transport system highly specific for cystine and glutamate. The underlying mechanism of cell death of cultured cells from the subtle gray (sut) mouse which contains an xCT null mutation remains elucidated. Our results show that the death of sut cells is likely caused by apoptosis mediated by c-Jun N-terminal kinase (JNK). The JNK activation triggers both a caspase-dependent (caspases-9 and -3) and an ER stress-mediated (eIF2 and CHOP) pathway to induce apoptosis. These findings suggest the possible pathways involved in the cell death of xCT-deficient cells. PMID- 18395004 TI - Catalytic mechanism of inulinase from Arthrobacter sp. S37. AB - Detailed catalytic roles of the conserved Glu323, Asp460, and Glu519 of Arthrobacter sp. S37 inulinase (EnIA), a member of the glycoside hydrolase family 32, were investigated by site-directed mutagenesis and pH-dependence studies of the enzyme efficiency and homology modeling were carried out for EnIA and for D460E mutant. The enzyme efficiency (k(cat)/K(m)) of the E323A and E519A mutants was significantly lower than that of the wild-type due to a substantial decrease in k(cat), but not due to variations in K(m), consistent with their putative roles as nucleophile and acid/base catalyst, respectively. The D460A mutant was totally inactive, whereas the D460E and D460N mutants were active to some extent, revealing Asp460 as a catalytic residue and demonstrating that the presence of a carboxylate group in this position is a prerequisite for catalysis. The pH dependence studies indicated that the pK(a) of the acid/base catalyst decreased from 9.2 for the wild-type enzyme to 7.0 for the D460E mutant, implicating Asp460 as the residue that interacts with the acid/base catalyst Glu519 and elevates its pK(a). Homology modeling and molecular dynamics simulation of the wild-type enzyme and the D460E mutant shed light on the structural roles of Glu323, Asp460, and Glu519 in the catalytic activity of the enzyme. PMID- 18395006 TI - Rehmannia inhibits adipocyte differentiation and adipogenesis. AB - Rehmannia glutinosa, a Traditional Chinese Medicine (TCM), has been used to increase physical strength. Here, we report that Rehmannia glutinosa extract (RE) inhibits adipocyte differentiation and adipogenesis. RE impairs differentiation of 3T3-L1 preadipocytes in a dose-dependent manner. At the molecular level, treatment with RE inhibits expression of the key adipocyte differentiation regulator C/EBPbeta, as well as C/EBPalpha and the terminal marker protein 422/aP2, during differentiation of preadipocytes into adipocytes. Additionally, RE inhibits the mitotic clonal expansion (MCE) process of adipocyte differentiation, and RE prevents localization of C/EBPbeta to the centromeres. RE also prevents high fat diet (HFD) induced weight gain and adiposity in rats. Taken together, our results indicate that Rehmannia glutinosa extract inhibits preadipocyte differentiation and adipogenesis in cultured cells and in rodent models of obesity. PMID- 18395007 TI - Osteogenic differentiation of human adipose tissue-derived stromal cells (hASCs) in a porous three-dimensional scaffold. AB - Recent studies have shown that liposuction aspirates from rat, rabbit, mouse, and human sources contain pluripotent adipose tissue-derived stromal cells (ASCs) that can differentiate into various mesodermal cell types, including osteoblasts, myoblasts, chondroblasts, and preadipocytes. To develop a research model for autologous bone tissue engineering, we isolated ASCs from human liposuction aspirates (hASCs) and induced their osteogenic differentiation in three dimensional poly(dl-lactic-co-glycolic acid) (PLGA) scaffolds. Human liposuction aspirates were proteolytically digested and centrifuged to obtain hASCs. After primary culture in control media and expansion to three passages, the cells were seeded in two-dimensional plates or three-dimensional PLGA scaffolds and cultured in osteogenic media for 4 weeks. In two-dimensional culture, osteogenesis was assessed by RT-PCR analysis of the osteogenic-specific bone sialoprotein mRNA, by alkaline phosphatase staining, and by von Kossa staining. In three-dimensional culture, osteogenesis was assessed by von Kossa and alizarine red S staining at 1, 2, and 4 weeks following osteogenic induction. hASCs incubated in two dimensional osteogenic media stained positively for alkaline phosphatase and with von Kossa stain after 2 weeks of differentiation. Expression of the osteogenesis specific bone sialoprotein gene was detected by RT-PCR after 2 weeks of differentiation. PLGA scaffolds seeded with hASCs showed multiple calcified extracellular matrix nodules by von Kossa and alizarine red S staining after 2 weeks of differentiation. In conclusion, the authors identified an osteogenic potential of hASCs and demonstrated osteogenic differentiation of hASCs into an osteogenic lineage in three-dimensional PLGA scaffolds. PMID- 18395008 TI - Gene expression patterns in glucose-stimulated podocytes. AB - To explore the mechanisms of podocyte injury under diabetic conditions, we performed an expression profile in glucose-stimulated podocytes. Differential gene expression profiles between conditionally immortalized mouse podocytes cultured in medium containing 5.6 and 30 mM glucose were measured with oligonucleotide microarrays. Of the genes identified, heme oxygenase-1, vascular endothelial growth factor-A, and thrombospondin-1 showed a consistently increased pattern, whereas angiotensin-converting enzyme-2 and peroxisomal proliferator activator receptor-gamma were down-regulated. These results were validated using real-time PCR and western blotting in podocytes, and with immunohistochemistry on renal tissues from streptozotocin-induced diabetic rats. Not only is this the first report of gene expression profiling of podocyte injury under diabetic conditions, but the identified genes are promising targets for future diabetes research. PMID- 18395009 TI - IRF-2 regulates NF-kappaB activity by modulating the subcellular localization of NF-kappaB. AB - Nuclear Factor-kappa B (NF-kappaB) is a transcription factor essential to the control of cell proliferation, survival, differentiation, immune response, and inflammation. Constitutive NF-kappaB activation has been observed in a broad variety of solid tumors and hematological malignancies, which suggests that NF kappaB signaling may perform a critical role in the development of human cancers. Interferon regulatory factor-2 (IRF-2), an antagonistic transcriptional repressor of IRF-1, evidences oncogenic potential, but little is currently known regarding the mechanism underlying the oncogenic activities of IRF-2. In this study, we report that IRF-2 recruits RelA/p65 transcription factors into the nucleus via physical interaction. While the nuclear recruitment of RelA by IRF-2 augments TNFalpha-induced NF-kappaB dependent transcription, the N-terminal truncated mutant form of IRF-2 inhibits the nuclear localization of RelA, and thus interferes with NF-kappaB activation. Furthermore, the knockdown of IRF-2 by IRF 2 siRNA attenuates TNFalpha-induced NF-kappaB dependent transcription by inhibiting the nuclear localization of RelA. Thus, these results show that IRF-2 regulates NF-kappaB activity via the modulation of NF-kappaB subcellular localization. PMID- 18395010 TI - Thyroid transcription factor-1 exhibits osmosensitive transcription in brain derived cell lines. AB - Thyroid transcription factor-1 (TTF-1) belongs to the Nkx family of homeodomain containing proteins and regulates expression of several important genes in the brain. Our previous studies showed that TTF-1 plays an important role in water homeostasis in the subfornical organ of rats and is involved in cerebrospinal fluid formation by regulation of aquaporin-1 transcription in the choroid plexus. In this study, we examined changes in TTF-1 transcription in response to hypertonicity using promoter assays. TTF-1 was synthesized in several osmosensitive regions of the rat brain. TTF-1 promoter activity was diminished by treatment with hypertonic solutions in a time- and dose-dependent manner in brain derived cell lines. Additionally, TTF-1 was involved in the regulation of angiotensinogen (Aogen) transcription under a hyperosmotic condition through specific binding domains in the Aogen promoter. These results suggest a possible role of TTF-1 in brain fluid homeostasis in response to changes in the osmotic environment. PMID- 18395011 TI - RIP and RALyase cleave the sarcin/ricin domain, a critical domain for ribosome function, during senescence of wheat coleoptiles. AB - Type-I ribosome-inactivating protein (RIP), which is found in many plants, catalyzes depurination of a specific adenine in the sarcin/ricin domain (SRD) of the large rRNA causing loss of ribosomal activity. Previously, we found a RNA apurinic site-specific lyase (RALyase) that catalytically cleaved the phosphodiester bond at the RIP-dependent depurination site by beta-elimination reaction. Here we show that both the RIP activity and RIP-RALyase-mediated cleavage of SRD in the cytoplasmic ribosome were induced at the late stage of senescence of wheat coleoptiles. Following this process, tissue death was observed. Furthermore, transgenic tobacco plants expressing glucocorticoid induced RIP developed senescence-like phenotype. Our results suggest that ribosome inactivation due to the cleavage of SRD by the inducible RIP and constitutively expressed RALyase may be a unique plant system that mediates programmed cell death at the late senescent stage. PMID- 18395012 TI - Inhibition of cholesterol transport into skin cells in cultures by phytosterol loaded microemulsion. AB - Cholesterol and plant phytosterols are lipophilic compounds solubilized by intestinal micelles in a competitive manner. In this work, we used radioactive cholesterol- and phytosterol-loaded oil-in-water microemulsions to follow their incorporation and mutual competition in HaCaT keratinocytes, SZ95 sebocytes, and skin pieces in cultures. Dynamic light scattering showed homogenous nanostructures of 10.5+/-1.5 nm diameter and cryo-transmission electron microscopy confirmed the presence of uniform spherical droplets of 7.0+/-1.0 nm diameter. Up to 320 nmol/ml of cholesterol can be solubilized and transported into cells with minimal toxic effect by 0.5 wt% nanodroplets in a cell medium. Phytosterols inhibit incorporation of cholesterol into cells, in vitro, at molar ratios (phytosterols/cholesterol) of 4 and above. The loaded nanodroplets accumulate in intracellular vesicles (presumably endosomes). No metabolic conversion of cholesterol or phytosterols was found in these cells, in vitro, after 24 h, at 37 degrees C. PMID- 18395013 TI - Human fucosyltransferase IX: specificity towards N-linked glycoproteins and relevance of the cytoplasmic domain in intra-Golgi localization. AB - The alpha3-fucosyltransferase IX (FUT9) catalyses the transfer of fucose in an alpha3 linkage onto terminal type II (Galbeta4GlcNAc) acceptors, the final step in the biosynthesis of the Lewisx (Lex) epitope, in neurons. In this work, FUT9 cloned from NT2N neurons and overexpressed in HeLa cells (FUT9wt), was found to efficiently fucosylate asialoerythropoietin (asialoEPO), and bovine asialofetuin, but not sialylated EPO. Analysis by HPAEC-PAD and MALDI/TOF-MS revealed predominantly mono-fucosylation by FUT9wt of type II di-, tri- and tetraantennary N-glycans with proximal fucose, with and without N-acetylactosamine repeats from asialoEPO. Minor amounts of difucosylated structures were also found. The results suggested that FUT9 could fucosylate Lex carrier-glycoproteins in neurons. Furthermore, FUT9wt was found to be activated by Mn2+ and it was capable of synthesizing Lea, although to a lesser extent than Lex and Ley. In vivo, HeLa cells transfected with FUT9wt expressed de novo Lex, as detected by immunofluorescence microscopy. FUT9 was found to be a trans-Golgi and trans-Golgi network (TGN) glycosyltransferase from confocal immunofluorescence co localization with the markers of the secretory pathway beta4 galactosyltransferase (trans-Golgi and TGN) and TGN-46 (TGN). Deletion of the cytoplasmic domain caused a shift to the cis-Golgi, thus suggesting that information for intra-Golgi localization is contained within the cytoplasmic domain. PMID- 18395014 TI - Quadruplex DNA crystal structures and drug design. AB - Crystallographic studies of G-quadruplex nucleic acids have resulted in a small group of structures to date. Their morphological and detailed conformational features are described here, emphasizing the stability of the G-tetrad core and the flexibility of loops, especially upon ligand binding. Implications for drug design are discussed, in the context of the druggability of both telomeric and non-telomeric quadruplex DNAs. PMID- 18395015 TI - Oxidative and nitrative protein modifications in Parkinson's disease. AB - Parkinson's disease (PD) is a complex neurodegenerative syndrome likely involving contributions from various factors in individuals including genetic susceptibility, exposure to environmental toxins, and the aging process itself. Increased oxidative stress appears to be a common causative aspect involved in the preferential loss of dopaminergic neurons in a region of the brain prominently affected by the disorder, the substantia nigra (SN). Loss of dopaminergic SN neurons is responsible for the classic clinical motor symptoms associated with PD. Several oxidative and nitrative posttranslational modifications (PTMs) have been identified on proteins pertinent to PD that may affect this or other aspects of disease progression. In this review, we discuss several examples of such PTMs to illustrate their potential consequences in terms of initiation or progression of PD neuropathophysiology. PMID- 18395016 TI - Deletion of apolipoprotein E gene modifies the rate of depletion of alpha tocopherol (vitamin E) from mice brains. AB - Our previous reports show that apolipoprotein E (apoE) influences the dynamics of alpha tocopherol (vitamin E) in brain. In this investigation, the patterns of depletion of alpha tocopherol from tissues of apoE deficient and wild type mice were compared after the animals were fed vitamin E deficient diets. Alpha tocopherol concentrations in specific regions of the brain and peripheral tissues at different times were determined by HPLC with electrochemical detection. ApoE deficiency significantly retarded the rate of depletion of alpha tocopherol from all regions of the brain. In addition, comparison of the rates of depletion of alpha tocopherol in both apoE deficient and wild type animals showed that cerebellum behaved differently from other areas such as cortex, hippocampus and striatum. This reinforces the uniqueness of cerebellum with regard to vitamin E biology. Patterns of depletion of tocopherol from peripheral tissues were different from brain. Serum tocopherol was higher in apoE deficient animals and remained higher than wild type during E deficiency. Depletion of liver tocopherol also tended to be unaffected by apoE deficiency. Our current and previous observations strongly suggest that apoE has an important role in modulating tocopherol concentrations in brain, probably acting in concert with other proteins as well. PMID- 18395019 TI - Results of a survey on current surgical smoke control practices. AB - Researchers at Duke University Medical Center, Durham, NC, conducted a simple, web-based survey of AORN members to evaluate surgical smoke control practices. Survey respondents from various medical specialties and facilities throughout North America indicated their facilities' level of compliance with established surgical smoke control measures. Survey results indicate that many facilities have not implemented best practices for protecting patients and health care workers from surgical smoke hazards, especially smoke created during electrosurgical, electrocautery, and diathermy procedures. PMID- 18395020 TI - Mandatory reporting and pay for performance: health care infections in the limelight. AB - Health care-associated infections (HAIs) have received national attention because of recent news stories in print and on television. many states have mandatory reporting requirements for hospital HAI rates, and as of October 1, 2008, Medicare no longer will pay for certain HAI-related costs. Collaboration among health care providers, patients, government agencies, professional and consumer groups, and payers and improved adherence to infection-prevention protocols can dramatically reduce the incidence of HAIs. PMID- 18395021 TI - Playing it safe: simulated team training in the OR. AB - Health care professionals are expected to make prompt, accurate decisions in life threatening emergency situations in the OR. Perioperative leaders are faced with the challenge of ensuring that OR team members are prepared to respond to potentially lethal emergencies, including rare and infrequent events. This article describes the importance of team training for all members of the OR team. The use of simulation is discussed as a tool to accomplish this goal. Steps for developing a simulation scenario are presented. PMID- 18395022 TI - The anxiety- and pain-reducing effects of music interventions: a systematic review. AB - Musical interventions have been used in health care settings to reduce patient pain, anxiety, and stress, although the exact mechanism of these therapies is not well understood. This article provides a systematic review of 42 randomized controlled trials of the effects of music interventions in perioperative settings. Music intervention had positive effects on reducing patients' anxiety and pain in approximately half of the reviewed studies. Further research into music therapy is warranted in light of the low cost of implementation and the potential ability of music to reduce perioperative patient distress. PMID- 18395023 TI - Registered nurse circulator legislation in Connecticut. AB - Developing regulations or legislation in every state to mandate an RN as the circulator in operating and invasive procedure rooms is critical to patient safety. Current regulatory language is ambiguous and may lead health care facility administrators to consider using licensed practical nurses, licensed vocational nurses, or surgical technologists in the circulator role. The State of Connecticut currently has no legislative or regulatory language regarding the RN as circulator, but some nurses are trying to change this. Communication with state legislators, has made it clear that legislators are in need of education about perioperative nursing and the circulator role. PMID- 18395025 TI - Placing patient safety first. PMID- 18395029 TI - A new model of ovarian carcinogenesis may influence early detection strategies. PMID- 18395030 TI - Early detection and treatment of ovarian cancer: shifting from early stage to minimal volume of disease based on a new model of carcinogenesis. AB - The goal of ovarian cancer screening is to detect disease when confined to the ovary (stage I) and thereby prolong survival. We believe this is an elusive goal because most ovarian cancer, at its earliest recognizable stage, is probably not confined to the ovary. We propose a new model of ovarian carcinogenesis based on clinical, pathological, and molecular genetic studies that may enable more targeted screening and therapeutic intervention to be developed. The model divides ovarian cancer into 2 groups designated type I and type II. Type I tumors are slow growing, generally confined to the ovary at diagnosis and develop from well-established precursor lesions so-called borderline tumors. Type I tumors include low-grade micropapillary serous carcinoma, mucinous, endometrioid, and clear cell carcinomas. They are genetically stable and are characterized by mutations in a number of different genes including KRAS, BRAF, PTEN, and beta catenin. Type II tumors are rapidly growing, highly aggressive neoplasms that lack well-defined precursor lesions; most are advanced stage at, or soon after, their inception. These include high-grade serous carcinoma, malignant mixed mesodermal tumors (carcinosarcomas), and undifferentiated carcinomas. The type II tumors are characterized by mutation of TP53 and a high level of genetic instability. Screening tests that focus on stage I disease may detect low-grade type I neoplasms but miss the more aggressive type II tumors, which account for most ovarian cancers. A more rational approach to early detection of ovarian cancer should focus on low volume rather than low stage of disease. PMID- 18395031 TI - Fibroids and reproductive outcomes: a systematic literature review from conception to delivery. AB - We examined the published relationship between uterine fibroids and reproductive outcomes. Submucosal fibroids had the strongest association with lower ongoing pregnancy rates, odds ratio, 0.5; 95% confidence interval, 0.3-0.8, primarily through decreased implantation. Cumulative pregnancy rates appeared slightly lower in patients with intramural fibroids 36.9% vs 41.1%, which may reflect biases in the literature; however, patients with intramural fibroids also experienced more miscarriages, 20.4% vs 12.9%. Adverse obstetric outcomes are rare and may reflect age or other differences in fibroid populations. Increased risk of malpresentation (odds ratio, 2.9; 2.6-3.2), cesarean (odds ratio, 3.7; 3.5-3.9), and preterm delivery (odds ratio, 1.5; 1.3-1.7) are reported; however, the incidence of labor dystocia was low (7.5%). There was no conclusive evidence that intramural or subserosal fibroids adversely affect fecundity. More prospective, controlled trials are needed to assess the effects of myomectomy. Good maternal and neonatal outcomes are expected in pregnancies with uterine fibroids. PMID- 18395033 TI - Pseudoamniotic band syndrome: a rare complication of monochorionic twins with fetofetal transfusion syndrome treated by laser coagulation. AB - OBJECTIVE: The purpose of this study was to assess the incidence and risk factors of limb constriction defects that are related to pseudoamniotic band syndrome (PABS) after selective fetoscopic laser surgery (FLS) in fetofetal transfusion syndrome (FFTS). STUDY DESIGN: All consecutive cases of FFTS that were treated by selective FLS between 1999 and 2006 were examined prospectively for PABS at the time of delivery. Incidence and characteristics of PABS were reported. Univariate analysis was conducted to look for potential risk factors of developing PABS. RESULTS: The 438 consecutive FFTS cases were treated at 15-26 weeks of gestation; PABS developed in 8 cases (1.8 %). The affected twin was always the former recipient. The diagnosis was made prenatally in 2 of 8 cases (25%). All cases survived the perinatal period. PABS affected fetal leg, arm, and foot in 3, 4, and 1 cases, respectively. In 5 (62.5%) and 7 (87.5%) cases, PABS occurred after premature rupture of membranes and intrauterine death of the donor, respectively. In 4 cases (50%), there was both premature rupture of membranes (PROM) and intrauterine fetal death; in 3 cases (37.5%), there was intrauterine fetal death alone, and in 1 case (12.5%), there was PROM alone. In the remaining 430 cases, PROM occurred in 62 cases (14.4%) and 66 cases (15.3%) within and after 3 weeks after surgery, respectively. PROM was significantly more frequent within the group that was complicated with PABS than within the rest of the cohort (P = .05). No maternal, fetal, or perioperative risk factor could be identified. CONCLUSION: Awareness and targeted serial ultrasound evaluation in this high-risk group may improve prenatal diagnosis, counseling, and management of PABS after FLS. PMID- 18395032 TI - Twin-to-twin transfusion syndrome: an antiangiogenic state? AB - OBJECTIVE: An imbalanced chronic blood flow between the donor and recipient twin through placental vascular anastomoses is the accepted pathophysiology of twin-to twin transfusion syndrome (TTTS). Vascular endothelial growth factor receptor-1 (VEGFR-1) mRNA is overexpressed only in the syncytiotrophoblast of the donor twin in some cases of TTTS. This study was conducted to determine maternal plasma concentrations of placental growth factor (PlGF), soluble VEGFR-1, and soluble endoglin (s-Eng) in monochorionic-diamniotic pregnancies with and without TTTS. STUDY DESIGN: This case-control study included monochorionic-diamniotic pregnancies between 16-26 weeks with and without TTTS. Maternal plasma concentrations of PlGF, sVEGFR-1, and s-Eng were determined with ELISA. A P value < .05 was considered statistically significant. RESULTS: Patients with TTTS had higher median plasma concentrations of s-Eng (14.8 ng/mL vs 7.8 ng/mL; P < .001) and sVEGFR-1 (6383.1 pg/mL vs 3220.1 pg/mL; P < .001]; and lower median plasma concentrations of PlGF (115.5 pg/mL vs 359.3 pg/mL; P = .002) than those without TTTS. CONCLUSION: We propose that an antiangiogenic state may be present in some cases of TTTS. PMID- 18395035 TI - The pharmaceutical industry: friend or foe? PMID- 18395034 TI - Nucleated red blood cells are a direct response to mediators of inflammation in newborns with early-onset neonatal sepsis. AB - OBJECTIVE: The objective of the study was to test the hypothesis that inflammation modulates fetal erythroblastosis and/or the release of nucleated red blood cells (NRBCs) independent of hypoxia or fetal stress. We sought to determine whether fetal inflammation is associated with an elevation in neonatal NRBC count in the setting of inflammation-associated preterm birth. STUDY DESIGN: The relationships between peripheral NRBC count, histological chorioamnionitis, umbilical cord interleukin (IL)-6, erythropoietin (EPO), cortisol, and acid-base status were analyzed in 68 preterm singletons, born to mothers who had an amniocentesis to rule out infection. Proteomic profiling of amniotic fluid identified presence of intraamniotic inflammation according to established parameters. NRBC counts were assessed within 1 hour of birth. Early-onset neonatal sepsis (EONS) was established based on hematological and microbiological indices. IL-6, EPO, and cortisol levels were measured by immunoassays. Fetal acid base status was determined within 10 minutes of delivery. Parametric or nonparametric statistics were used. RESULTS: Fetuses with EONS (n = 19) were delivered at earlier gestational ages (mean +/- SD: 27.1 +/- 2.8 weeks, P = .001) and more often by mothers with intraamniotic inflammation (P = .022) and histological chorioamnionitis (P < .001). Neonates with EONS had higher absolute NRBC counts (P = .011). NRBC counts were directly correlated with cord blood IL-6 levels (P < .001) but not with EPO, cortisol or parameters of acid-base status levels regardless of EONS status. These relationships remained following correction for gestational age, diabetes, intrauterine growth restriction, and steroid exposure. CONCLUSION: In the setting of inflammation-associated preterm birth and in the absence of hypoxia, elevations in NRBCs in the early neonatal period may be a direct response of exposure to inflammatory mediators in utero. PMID- 18395036 TI - Fetal outcome in motor-vehicle crashes: effects of crash characteristics and maternal restraint. AB - OBJECTIVE: This project was undertaken to improve understanding of factors associated with adverse fetal outcomes of pregnant occupants involved in motor vehicle crashes. STUDY DESIGN: In-depth investigations of crashes involving 57 pregnant occupants were performed. Maternal and fetal injuries, restraint information, measures of external and internal vehicle damage, and details about the crash circumstances were collected. Crash severity was calculated using vehicle crush measurements. Chi-square analysis and logistic regression models were used to determine factors with a significant association with fetal outcome. RESULTS: Fetal outcome is most strongly associated with crash severity (P < .001) and maternal injury (P = .002). Proper maternal belt-restraint use (with or without airbag deployment) is associated with acceptable fetal outcome (odds ratio = 4.5, P = .033). Approximately half of fetal losses in motor-vehicle crashes could be prevented if all pregnant women properly wore seat belts. CONCLUSION: Higher crash severity, more severe maternal injury, and lack of proper seat belt use are associated with a higher risk of adverse fetal outcome. These results strongly support recommendations that pregnant women use properly positioned seatbelts. PMID- 18395037 TI - Vaginal birth after cesarean: clinical risk factors associated with adverse outcome. AB - OBJECTIVE: The objective of the study was to identify vaginal birth after cesarean (VBAC) success rates and maternal and neonatal complication rates for selected antenatal conditions. STUDY DESIGN: This was a population-based cohort study using administrative discharge data for women delivering in California hospitals during 2002. RESULTS: Among 41,450 women, 29.72% (12,320 of 41,450) had maternal, fetal, or placental conditions complicating pregnancy. Attempted VBAC rates and VBAC success rates varied widely by these clinical condition, ranging from 10% to 73%. The VBAC success rate for low-risk women (no conditions) was 73.76% vs 50.31% for high-risk women (at least 1 condition), P < .0001. Absolute rates of maternal and neonatal complications were low (less than 1-2%), and the rate of adverse events was higher in the high-risk clinical group as compared with the low-risk clinical group. CONCLUSION: Variation in rates of VBAC success and childbirth morbidities can be partially attributed to clinical factors complicating pregnancy. Women without such conditions show improved VBAC success and fewer maternal and neonatal complications. PMID- 18395038 TI - Gene expression profiling of human oocytes at different maturational stages and after in vitro maturation. AB - OBJECTIVE: The purpose of this study was to catalog genes expressed in human oocytes at germinal vesicle (GV) or metaphase II (MII) stage and to compare gene profiles between oocytes matured in vivo (in vivo-MII) and in vitro (IVM-MII). STUDY DESIGN: University-based research utilizing unfertilized oocytes analyzed for > 29,000 genes with RNA amplification and microarray. RESULTS: GV, in vivo MII, and IVM-MII oocytes expressed 12,219, 9735, and 8510 genes, respectively. There was extensive overlap among the 3 groups, but also some significant differences. In particular, in vivo-MII and IVM-MII oocytes shared very similar patterns of gene expression. However, some immature patterns of expression, reminiscent of GVs, persisted in IVM-MIIs. CONCLUSION: In vitro maturation is an attractive strategy for IVF treatment; however, current IVM methods produce oocytes that perform poorly in the context of IVF. Data from the current study suggest that although IVM-MII oocytes closely resemble in vivo-MII oocytes for cellular pathways related to nuclear maturity, several pathways associated with cytoplasmic functions continue to be expressed in an immature manner. Additionally, IVM-MII oocytes have differences in the expression of genes related to cellular storage and homeostasis. Differentially expressed genes/pathways provide clues for the optimization of IVM techniques. PMID- 18395039 TI - Is there a therapeutic impact to regional lymphadenectomy in the surgical treatment of endometrial carcinoma? AB - OBJECTIVE: The objective of the study was to evaluate whether surgical removal of regional lymph nodes influences the overall survival of women with endometrial carcinoma. STUDY DESIGN: All patients with endometrioid-type endometrial cancer treated at our institution between January 1993 and December 2004 were reviewed. The Classification and Regression Tree (CART) method, a form of recursive partitioning, was used. RESULTS: In all, 1035 patients were evaluated. International Federal of Gynecology and Obstetrics stage included the following: stage I, 824; stage II, 65; stage III, 109; and stage IV, 37. Lymph nodes were removed in 524 patients (51%). The median number of nodes removed was 16. Using the CART hierarchically, stage, age, adjuvant therapy, and the removal of 10 lymph nodes or more emerged as predictors of overall survival. CONCLUSION: This study emphasizes the importance of lymph node dissection in endometrial cancer. Lymph node dissection is essential for accurate surgical staging, which remains the most important prognostic factor. In addition to well-known clinicopathologic risk factors for survival, the removal of 10 or more regional lymph nodes was associated with improved overall survival in lower-stage, older patients who received no adjuvant therapy or brachytherapy only. PMID- 18395040 TI - Epidemiology of low-grade serous ovarian cancer. AB - OBJECTIVE: The objective of the study was to determine whether the epidemiology of low-grade ovarian serous cancers is distinct from that of high-grade ovarian serous cancers. STUDY DESIGN: The National Cancer Institute's Surveillance Epidemiology and End Results (SEER) program reports incidence, and survival, data from representative population based cancer registries. The SEER data are analyzed to compare the descriptive epidemiologies of low- and high-grade lesions. RESULTS: Mean survival (99 vs 57 months) and age (55.5 vs 62.6 years) trends in annual incidence rate (-3.8% vs 1.4%), rate ratio of advanced to early disease (1.9 vs 10.2), rate ratio of post- to premenopausal incidence (5.0 vs 13.0). and difference in average age between advanced and early stage (none vs 2.5 years) are significantly different for patients with low-grade, compared with high-grade, disease. CONCLUSION: Epidemiology of low-grade tumors appears sufficiently different from that of high-grade lesions to support the concept that low-grade ovarian serous cancers constitute a distinct clinical, and perhaps biologic, entity. PMID- 18395041 TI - Reliability and validity of reflection exercises for obstetrics and gynecology residents. AB - OBJECTIVE: Self-evaluation is an essential skill throughout a physician's career, and reflection is thought to be a necessary mechanism for effective self evaluation. The aim of our study was to establish the reliability and validity of structured assessments of critical self-reflection. STUDY DESIGN: Thirty-two residents completed 6 exercises that were scored from 0 (no description of event) to 6 (deep reflection). We calculated interrater and internal consistency reliability for the exercises and compared scores by postgraduate year and with other competency assessments. RESULTS: Residents completed 183 reflections. Interrater reliability was 0.89. Surgical skill reflections scored highest (score, 3.2 +/- 0.91 [SD]). Five exercises had adequate internal consistency reliability (0.62). Senior residents received higher reflection scores than junior residents; the magnitude of difference was similar for other competency measures and not statistically significant. Reflection scores were correlated with professionalism and communication skill assessments (score, 0.36-0.37; P < .01) but not with medical knowledge. CONCLUSION: Self-reflection can be assessed reliably with scored exercises that demonstrate concurrent validity with other assessments. We encourage further research that should include multiple training programs to further evaluate our approach for the assessment of reflection in postgraduate education. PMID- 18395042 TI - Protection against cisplatin-induced ovarian damage by the antioxidant sodium 2 mercaptoethanesulfonate (mesna) in female rats. AB - OBJECTIVE: The hypothesis was that the administration of the antioxidant mesna (sodium 2-mercaptoethanesulfonate) during chemotherapy would protect ovaries against follicular damage. STUDY DESIGN: Sprague-Dawley rats were treated with saline solution, mesna-plus-cisplatin, or cisplatin. Immunohistochemistry was used to evaluate the Mullerian inhibiting substance (MIS) positive follicles. Serum and ovarian MIS were measured with enzyme-linked immunosorbent assay and Western blot analysis, respectively. Apoptosis in ovaries was studied by terminal deoxynucleotidyl transfer biotin-d UTP nick end labeling (TUNEL) assay. RESULT: Immunofluorescence staining for MIS was higher in preantral follicles in the mesna-plus-cisplatin group. The ovarian and serum MIS levels were higher in the mesna-plus-cisplatin than in the cisplatin alone group. There were no differences statistically in the TUNEL and the ovarian cyst analyses. CONCLUSION: Mesna, which was used at the time of cisplatin administration, protected ovaries against damage. The data that are presented challenge the existing clinical paradigm that gonadotropin-releasing hormone agonists represent the only medical method for the protection of ovaries during chemotherapy. Alternative medical means to protect ovaries during chemotherapy may be achievable. PMID- 18395043 TI - Early preterm birth: association between in utero exposure to acute inflammation and severe neurodevelopmental disability at 6 years of age. AB - OBJECTIVE: The purpose of this study was to determine the association between in utero exposure to acute inflammation and long-term major neurodevelopmental disability at age 6 years among children born prior to 32 weeks' gestation. STUDY DESIGN: This was a follow-up investigation of a cohort of maternal-infant dyads delivered between 23 and < 32 weeks' gestation. Surviving infants (and their mothers or caregivers) underwent a battery of psychological and neurodevelopmental tests between 5 and 8 years of age. Pregnancy and neonatal data were analyzed among children with versus those without major neurodevelopmental disability (including IQ < 70 [n = 41], cerebral palsy [CP, n = 11], and a composite major disability [n = 52]). RESULTS: A total of 261 (70%) of the 375 maternal-infant dyads with surviving children were successfully recruited and evaluated at 6.8 +/- 0.7 years. Mean delivery gestational age (GA) and birthweight were 28.8 +/- 2.2 weeks and 1163 +/- 382 g, respectively. Neither surrogate indicators for nor direct markers of in utero exposure to acute inflammation were significantly associated with severe adverse outcomes. Delivery GA was significantly associated with outcome. Logistic regression indicated that each increasing gestational week was associated with a significantly decreased risk of an IQ < 70 (OR 0.75, 95% CI 0.6-0.9). An average 1.9 point increase in IQ at 6 years of age was observed per gestational week gained (23 to 32 weeks). Periventricular leukomalacia was associated with a 9.6 point mean deficit in IQ. The perceptive vocabulary scores (IQ proxy) of primary caregivers were significantly lower among children with an IQ < 70 vs > or = 70 (87.5 +/- 11.5 vs 92.1 +/- 11.2, P = .016). CONCLUSION: Among children born between 23 and 32 weeks' gestation, neonatal complications, GA at delivery, and caregiver IQ, but not in utero exposure to acute inflammation, were associated with increased risk of severe adverse neurodevelopmental outcomes at age 6 years. PMID- 18395044 TI - Prediction of preterm birth in symptomatic women using decision tree modeling for biomarkers. AB - OBJECTIVE: The objective of the study was to use recursive partitioning (RP) to identify gestational age-specific and threshold values for infectious and endocrine biomarkers of imminent delivery. STUDY DESIGN: RP was developed using a previously collected data set and then applied to a prospectively collected cohort of women in threatened preterm labor. Predictors of preterm birth were considered, including white blood cell count (WBC), corticotrophin-releasing hormone (CRH), cortisol, and maternal age. RESULTS: At 22-27 weeks' gestation, WBC of greater than 12,000/mL was the most accurate predictor of delivery within 48 hours; at 28-31 weeks' gestation, CRH of greater than 684 pg/mL was the most accurate predictor; and at 32-26 weeks' gestation, CRH and maternal age were the most important variables. CONCLUSIONS: These results indicate that maternal WBC greater than 12,000/mL prior to 28 weeks' gestation and CRH beyond 28 weeks are the most accurate biomarkers in predicting preterm birth within 48 hours. RP assists in establishing clinically relevant and gestational age-specific threshold levels for these variables. PMID- 18395045 TI - Diminished survival of human cytotrophoblast cells exposed to hypoxia/reoxygenation injury and associated reduction of heparin-binding epidermal growth factor-like growth factor. AB - OBJECTIVE: The antiapoptotic action of heparin-binding epidermal growth factor (HBEGF)-like growth factor and its regulation by O(2) constitutes a key factor for trophoblast survival. The hypothesis that cytotrophoblast survival is compromised by exposure to hypoxia-reoxygenation (H/R) injury, which may contribute to preeclampsia and some missed abortions, prompted us to investigate HBEGF regulation and its role as a survival factor during H/R in cytotrophoblast cells. STUDY DESIGN: A transformed human first-trimester cytotrophoblast cell line HTR-8/SVneo was exposed to H/R (2% O(2) followed by 20% O(2)) and assessed for HBEGF expression and cell death. RESULTS: Cellular HBEGF declined significantly within 30 minutes of reoxygenation after culture at 2% O(2). H/R significantly reduced proliferation and increased cell death when compared with trophoblast cells cultured continuously at 2% or 20% O(2). Restoration of cell survival also was achieved by adding recombinant HBEGF during reoxygenation. HBEGF inhibited apoptosis through its binding to either human epidermal receptor (HER)-1 or HER4, its cognate receptors. CONCLUSION: These results provide evidence that cytotrophoblast exposure to H/R induces apoptosis and decreased cell proliferation. HBEGF accumulation is diminished under these conditions, whereas restoration of HBEGF signaling improves trophoblast survival. PMID- 18395046 TI - Mechanical homeostasis is altered in uterine leiomyoma. AB - OBJECTIVE: Uterine leiomyoma produce an extracellular matrix (ECM) that is abnormal in its volume, content, and structure. Alterations in ECM can modify mechanical stress on cells and lead to activation of Rho-dependent signaling and cell growth. Here we sought to determine whether the altered ECM that is produced by leiomyoma was accompanied by an altered state of mechanical homeostasis. STUDY DESIGN: We measured the mechanical response of paired leiomyoma and myometrial samples and performed immunogold, confocal microscopy, and immunohistochemical analyses. RESULTS: Leiomyoma were significantly stiffer than matched myometrium. The increased stiffness was accompanied by alteration of the ECM, cell shape, and cytoskeleton in leiomyoma, compared with myometrial samples from the same uterus. Levels of AKAP13, a protein that is known to activate Rho, were increased in leiomyoma compared to myometrium. AKAP13 was associated with cytoskeletal filaments of immortalized leiomyoma cells. CONCLUSION: Leiomyoma cells are exposed to increased mechanical loading and show structural and biochemical features that are consistent with the activation of solid-state signaling. PMID- 18395048 TI - Predicting congenital cytomegalovirus infection: Guerra et al. AB - The article below summarizes a roundtable discussion of a study published in this issue of the Journal in light of its methodology, relevance to practice, and implications for future research. Article discussed: Guerra B, Simonazzi G, Puccetti C, et al. Ultrasound prediction of symptomatic congenital cytomegalovirus infection. Am J Obstet Gynecol 2008;198:380.e1-380.e7. The full discussion appears at www.AJOG.org, page e1-e3. PMID- 18395049 TI - A dangling development. PMID- 18395047 TI - Impact of vessel maturation on antiangiogenic therapy in ovarian cancer. AB - OBJECTIVE: The purpose of this study was to examine the functional and therapeutic significance of pericytes in ovarian cancer vasculature. STUDY DESIGN: Tumor vessel morphologic condition and efficacy of endothelial and pericyte targeting were examined with the use of in vivo ovarian cancer models. The expression of platelet-derived growth factor (PDGF) ligands and receptors was examined in endothelial, pericyte-like, and ovarian cancer cells. RESULTS: Relative to normal vessels, tumor vasculature was characterized by loosely attached pericytes in reduced density. PDGF-BB was expressed predominantly by the endothelial and cancer cells, whereas PDGFRbeta was present in pericyte-like cells. PDGF-BB significantly increased the migration of and VEGF production by pericyte-like cells; PDGFRbeta blockade abrogated these effects. Dual VEGF (VEGF Trap) and PDGF-B (PDGF-Trap) targeted therapy was more effective in inhibiting in vivo tumor growth than either agent alone. CONCLUSION: Aberrations in the tumor microenvironment contribute to endothelial cell survival. Strategies that target both endothelial cells and pericytes should be considered for clinical trials. PMID- 18395050 TI - Race as a social construct: the genetic fallacy. PMID- 18395051 TI - Chemometric methods for evaluation of chromatographic separation quality from two way data--a review. AB - Most traditional chromatographic separation criteria or response functions are defined on chromatograms recorded by single-channel detectors, e.g. a spectrometer measuring the absorbance at a single wavelength or a thermal conductivity detector. When the peaks are seriously overlapped, usually there is a lack of the information concerning the total number of chemical components, overlap degree of the peaks and peak purity. Such information characterizes some crucial aspects of separation process and lack of it will lead to an inaccurate and misleading evaluation of separation quality as well as some computational ambiguity for many traditional response functions. In contrast, hyphenated chromatography-(multi-channel) spectroscopy instruments together with chemometric methods will largely increase the information content available in chromatographic detection. Such information, if properly used, can cast a new light on evaluation of chromatographic separation quality. The main objective of this article is to review chemometric methods devoted to estimation of the number of chemical components, determination of elution sequence and assessment of peak purity. Some newly defined response functions or separation criteria based on extracted information by chemometric methods are also introduced. The methods reviewed are limited to those for treating two-way data obtained by hyphenation of high-performance liquid chromatography with multi-channel detectors. We prefer to provide a comprehensive view of such methods rather than present a full list of all the methods developed. Further details of some important methods are touched upon in favor of employment and understanding of them by researchers not very familiar with chemometrics. PMID- 18395052 TI - Titanium dioxide determination in sunscreen by energy dispersive X-ray fluorescence methodology. AB - Nowadays there are many sun-protection cosmetics incorporating chemical and/or physical UV filters as active ingredients and there are no official methods to determine these kinds of compounds in sunscreen cosmetics. The objective of this work is to estimate TiO(2) concentration, without sample preparation, employing a portable energy dispersive X-ray fluorescence (EDXRF), aiming to estimate the sun protection factor (SPF) due to the physical barrier in sunscreen composition, and also identify the metals present in the samples. A portable EDXRF system was used for the analysis of fifteen commercial samples. It was also prepared three formulations estimated in FPS-30 using TiO(2) at 5%. Quantification was performed using calibration curves with standards from 1 to 30%. The physical barrier contribution in the SPF, associated to Ti concentration, was determined for all samples. The presence of some elements, like K, Zn, Br and Sr was detected in the sunscreen, identifying chemical elements that were not cited in the formulations. Three commercial samples were analyzed for total SPF determination and the result shows that the measured value is 10% lower than the nominal one. PMID- 18395053 TI - A new strategy for solving matrix effect in multivariate calibration standard addition data using combination of H-point curve isolation and H-point standard addition methods. AB - This work presents a new and simple strategy for solving matrix effects using combination of H-point curve isolation method (HPCIM) and H-point standard addition method (HPSAM). The method uses spectrophotometric multivariate calibration data constructed by successive standard addition of an analyte into an unknown matrix. By successive standard addition of the analyte, the concentrations of remaining components (interferents) remain constant and therefore give constant cumulative spectrum for interferents in the unknown mixture. The proposed method firstly extracts such spectrum using H-point curve isolation method and then applies the obtained cumulative interferents spectrum for determination of analyte by H-point standard addition method. In order to evaluate the applicability of the method a simulated as well as several experimental data sets were tested. The method was then applied to the determination of paracetamol in pharmaceutical tablets and copper in urine samples and in a copper alloy. PMID- 18395054 TI - Electrogenerated chemiluminescence: an oxidative-reductive mechanism between quinolone antibiotics and tris(2,2'-bipyridyl)ruthenium(II). AB - The cyclic voltammetry and electrogenerated chemiluminescent (ECL) reactions of a series of quinolone and fluoroquinolone antibiotics were investigated in a flow injection analysis (FIA) system. 7-Piperazinyl fluoroquinolone antibiotics were found to participate as a coreactant in an oxidative-reductive ECL mechanism with tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)(3)(2+)) as the luminescent reagent. The reaction mechanism was investigated in order to understand and optimize the processes leading to light emission. The optimal conditions included a solution pH approximately 7 at a flow rate of 3.0 mL min(-1) with no added organic modifier and application of 1.2V vs. a Pt quasi-reference electrode (QRE). Fluoroquinolones containing a tertiary distal nitrogen on the piperazine ring, such as enrofloxacin and ofloxacin, reacted to produce more intense ECL than those with a secondary nitrogen, such as ciprofloxacin and norfloxacin. The method linear range, precision, detection limits, and sensitivity for the detection of enrofloxacin and ciprofloxacin were compared to that of tripropylamine. The method was applied to the determination of the ciprofloxacin content in a pharmaceutical preparation. The assay is discussed in terms of its analytical figures of merit, ease of use, speed, accuracy and application to pharmaceutical samples. PMID- 18395055 TI - Label-free aptasensor for platelet-derived growth factor (PDGF) protein. AB - A label-free aptasensor for platelet-derived growth factor (PDGF) protein is reported. The aptasensor uses mixed self-assembled monolayers (SAMs) composed of a thiol-modified PDGF binding aptamer and 6-mercaptohexanol (MCH) on a gold electrode. The SAMs were characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV) before and after binding of the protein using [Fe(CN)(6)](3-/4-), a redox marker ion as an indicator for the formation of a protein-aptamer complex. The CVs at the PDGF modified electrode showed significant differences, such as changes in the peak currents and peak-to-peak separation, before and after binding of the target protein. The EIS spectra, in the form of Nyquist plots, were analyzed with a Randles circuit while the electron transfer resistance R(ct) was used to monitor the binding of the target protein. The results showed that, without any modification to the aptamer, the target protein can be recognized effectively at the PDGF binding aptamer SAMs at the electrode surface. Control experiments using non-binding oligonucleotides assembled at the electrode surfaces also confirmed the results and showed that there was no formation of an aptamer-protein complex. The DPV signal at the aptamer functionalized electrode showed a linearly decreased marker ion peak current in a protein concentrations range of 1-40 nM. Thus, label-free detection of PDGF protein at an aptamer modified electrode has been demonstrated. PMID- 18395056 TI - A combination of statistical and analytical evaluation methods as a new optimization strategy for the quantification of pharmaceutical residues in sewage effluent. AB - In this study, a new solid-phase microextraction (SPME) method for simultaneous extraction of pharmaceutical compounds with acidic and basic characteristics (ibuprofen, fenoprofen, diclofenac, diazepam and loratadine) from residual water samples is proposed. In this procedure, the extraction is processed using two distinct sample pH values. The extraction is begun at pH 2.5 to promote the sorption of acidic pharmaceuticals and after 35 min the sample pH is changed to 7.0 by adding 0.4 mol L(-1) disodium hydrogenphosphate, so that the basic compounds can be sorbed by the fiber (20 min). The pH change is performed without interruption of the extraction process. A comparison between the proposed method and the SPME method applied to each group of the target compounds was performed. Gas chromatography coupled to mass spectrometry was used for separation and detection of analytes. The extraction conditions for the three methods were optimized using full factorial experimental design, response surface through a Doehlert matrix and central composite design. Limits of detection (0.02-0.43 microg L(-1)) and correlation coefficients (0.9970-0.9998) were determined for the three methods. The proposed extraction procedure was applied to samples of sewage treatment plant effluent and untreated wastewater. Recovery and relative standard deviation values ranged from 67 to 116% and 4.6 to 14.5%, respectively, for all compounds studied. Modification of sample pH during the extraction procedure was shown to be an excellent option for all of the compounds and may be extended to the simultaneous extraction of other compounds with different acid base characteristics. PMID- 18395057 TI - Flow methodology for methanol determination in biodiesel exploiting membrane based extraction. AB - A methodology based in flow analysis and membrane-based extraction has been applied to the determination of methanol in biodiesel samples. A hydrophilic membrane was used to perform the liquid-liquid extraction in the system with the organic sample fed to the donor side of the membrane and the methanol transfer to an aqueous acceptor buffer solution. The quantification of the methanol was then achieved in aqueous solution by the combined use of immobilised alcohol oxidase (AOD), soluble peroxidase and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). The optimization of parameters such as the type of membrane, the groove volume and configuration of the membrane unit, the appropriate organic solvent, sample injection volume, as well as immobilised packed AOD reactor was performed. Two dynamic analytical working ranges were achieved, up to 0.015% and up to 0.200% (m/m) methanol concentrations, just by changing the volume of acceptor aqueous solution. Detection limits of 0.0002% (m/m) and 0.007% (m/m) methanol were estimated, respectively. The decision limit (CCalpha) and the detection capacity (CCbeta) were 0.206 and 0.211% (m/m), respectively. The developed methodology showed good precision, with a relative standard deviation (R.S.D.) <5.0% (n=10). Biodiesel samples from different sources were then directly analyzed without any sample pre-treatment. Statistical evaluation showed good compliance, for a 95% confidence level, between the results obtained with the flow system and those furnished by the gas chromatography reference method. The proposed methodology turns out to be more environmental friendly and cost effective than the reference method. PMID- 18395058 TI - Analysis of alkaloids in Coptis chinensis Franch by accelerated solvent extraction combined with ultra performance liquid chromatographic analysis with photodiode array and tandem mass spectrometry detections. AB - A new method based on accelerated solvent extraction (ASE) followed by ultra performance liquid chromatography (UPLC) analysis has been developed for the identification and quantification of major alkaloids in extracts of Coptis chinensis Franch. The UPLC system consisted of a dual detection system of photodiode array detector (PDA) and positive ion electrospray ionization-tandem mass spectrometry (ESI-MS/MS) in sequential configuration. The operational parameters of ASE including extraction solvent, extraction temperature, static extraction time and extraction cycles were optimized. UPLC analysis was performed on an ACQUITY UPLC BEH C(18) column eluted by a mobile phase of acetonitrile spiked with a buffer solution consisting of 0.50% acetic acid and 20 mmol L(-1) ammonium acetate. A tandem quadrupole spectrometer operating in either full scan mode or in MS/MS mode for multiple reaction monitoring (MRM) was used for the identification and quantitative analysis of eight major alkaloids in C. chinensis Franch extracts. The samples were also analyzed on a high-performance liquid chromatography-electrospray ionization-time-of-flight mass spectrometry (HPLC-ESI TOF-MS) system to confirm the identification results. Three of the eight major alkaloids, berberine, palmatine and jatrorrhizine were quantified by UPLC-PDA and UPLC-MS/MS. The results indicated that both UPLC-PDA and UPLC-MS/MS methods were simple, sensitive and reliable for the determination of alkaloids in C. chinensis Franch. Seven Huanglian samples from different locations were analyzed using the established methods. UPLC fingerprints based on the distribution of the eight major alkaloids can serve as a rapid and reliable method for the authentication and quality evaluation of traditional Chinese medicine (TCM) herbs. PMID- 18395059 TI - Chemometric determination of arsenic and lead in untreated powdered red paprika by diffuse reflectance near-infrared spectroscopy. AB - It has been evaluated the potential of near-infrared (NIR) diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) as a way for non-destructive measurement of trace elements at microg kg(-1) level in foods, with neither physical nor chemical pre-treatment. Predictive models were developed using partial least-square (PLS) multivariate approaches based on first-order derivative spectra. A critical comparison of two spectral pre-treatments, multiplicative signal correction (MSC) and standard normal variate (SNV) was also made. The PLS models built after using SNV provided the best prediction results for the determination of arsenic and lead in powdered red paprika samples. Relative root-mean-square error of prediction (RRMSEP) of 23% for both metals, arsenic and lead, were found in this study using 20 well characterized samples for calibration and 13 additional samples as validation set. Results derived from this study showed that NIR diffuse reflectance spectroscopy combined with the appropriate chemometric tools could be considered as an useful screening tool for a rapid determination of As and Pb at concentration level of the order of hundred microg kg(-1). PMID- 18395060 TI - Simultaneous analysis of riboflavin and aromatic amino acids in beer using fluorescence and multivariate calibration methods. AB - The study demonstrates an application of the front-face fluorescence spectroscopy combined with multivariate regression methods to the analysis of fluorescent beer components. Partial least-squares regressions (PLS1, PLS2, and N-way PLS) were utilized to develop calibration models between synchronous fluorescence spectra and excitation-emission matrices of beers, on one hand, and analytical concentrations of riboflavin and aromatic amino acids, on the other hand. The best results were obtained in the analysis of excitation-emission matrices using the N-way PLS2 method. The respective correlation coefficients, and the values of the root mean-square error of cross-validation (RMSECV), expressed as percentages of the respective mean analytic concentrations, were: 0.963 and 14% for riboflavin, 0.974 and 4% for tryptophan, 0.980 and 4% for tyrosine, and 0.982 and 19% for phenylalanine. PMID- 18395061 TI - A novel application of nylon membranes to the luminescent determination of benzo[a]pyrene at ultra trace levels in water samples. AB - Very simple and highly sensitive methods are presented for the determination of benzo[a]pyrene, one of the most carcinogenic polycyclic aromatic hydrocarbons (PAHs). The approaches are based on solid-phase extraction of the analyte on a nylon membrane via a syringe procedure, and its fluorescent or phosphorescent determination on the solid surface. While the native fluorescence of benzo[a]pyrene retained on a nylon surface is measured directly, room-temperature phosphorescence is induced by spotting a few microlitres of thallium(I) nitrate solution on the surface (heavy-atom effect). An enhancement of the phosphorescence signal was corroborated when the measurements were carried under a nitrogen atmosphere. The analytical figures of merit obtained under the best experimental conditions demonstrate the capability of detecting benzo[a]pyrene at a sub-parts-per-trillion (sub-ng L(-1)) level. The potential interference from other common PAHs and also from different metal ions was studied. The feasibility of determining benzo[a]pyrene in real samples was successfully evaluated through the analysis of spiked tap, underground and mineral water samples of different origins. Recoveries obtained from spiked river waters were successfully compared with those provided by a reference method, through rigorous statistical analysis. PMID- 18395062 TI - Profiling of 19-norsteroid sulfoconjugates in human urine by liquid chromatography mass spectrometry. AB - 19-Nortestosterone (nandrolone) major metabolites in human urine are excreted as sulfoconjugated and glucuroconjugated forms. A sensitive and selective liquid chromatography/tandem mass spectrometry (LC/MS/MS) method in negative ESI mode was developed for direct quantification of 19-norandrosterone sulfate (19-NAS) and 19-noretiocholanolone sulfate (19-NES). For both sulfoconjugates, the [M-H]( ) ion at m/z 355 and the fragment ion at m/z 97 were used as the precursor and product ions, respectively. The purification method involved a complete and rapid separation of sulfates and glucuronides in two extracts after loading the sample on a weak anion exchange solid phase extraction support (SPE Oasis WAX). Then, sulfates were separated by LC (Uptisphere ODB, 150 mm x 3.0 mm, 5 microm) and analyzed on a linear trap and a triple quadrupole mass spectrometer. The lower limit of detection (LLOD) and lowest limit of quantification (LLOQ) were of 100 pg mL(-1) and 1 ng mL(-1), respectively. Assay validation demonstrated good performances in terms of trueness (92.0-104.9%), repeatability (0.6-7.2%) and intermediate precision (1.3-10.8%) over the range of 1-2500 ng mL(-1). Finally, 19-NAS and 19-NES in urine samples collected after intake of 19 norandrostenedione (nandrolone precursor) were quantified. This assay may be easily implemented to separate glucuronide and sulfate steroids from urine specimens prior to quantification by LC/MS/MS. PMID- 18395063 TI - Electronic clinical challenges and images in GI. Migration of a gastric bezoar. PMID- 18395064 TI - Electronic clinical challenges and images in GI. Meckel's diverticulum. PMID- 18395066 TI - Gaps in adopting conflicts of interest policies among u.s. Medical schools. PMID- 18395067 TI - FDA set to tighten guidelines on industry dissemination of information on unapproved uses of medical products. PMID- 18395070 TI - AGA PAC: an essential tool to advance gastroenterology. PMID- 18395071 TI - Women in gastroenterology: exciting times and trends. PMID- 18395072 TI - Endoscopic imaging of angiogenesis in vivo. PMID- 18395073 TI - Clinical challenges and images in GI. Esophagopleural fistula. PMID- 18395074 TI - Clinical challenges and images in GI. Intestinal Waldenstrom macroglobulinemia. PMID- 18395075 TI - Gastric cancer risk in patients with premalignant gastric lesions: a nationwide cohort study in the Netherlands. AB - BACKGROUND & AIMS: A cascade of precursor lesions (eg, atrophic gastritis, intestinal metaplasia, and dysplasia) precedes most gastric adenocarcinomas. Quantification of gastric cancer risk in patients with premalignant gastric lesions is unclear, however. Consequently, endoscopic surveillance is controversial, especially in Western populations. METHODS: To analyze current surveillance practice and gastric cancer risk in patients with premalignant gastric lesions, all patients with a first diagnosis between 1991 and 2004 were identified in the Dutch nationwide histopathology registry (PALGA); follow-up data were evaluated until December 2005. RESULTS: In total, 22,365 (24%) patients were diagnosed with atrophic gastritis, 61,707 (67%) with intestinal metaplasia, 7616 (8%) with mild-to-moderate dysplasia, and 562 (0.6%) with severe dysplasia. Patients with a diagnosis of atrophic gastritis, intestinal metaplasia, or mild to-moderate dysplasia received re-evaluation in 26%, 28%, and 38% of cases, respectively, compared with 61% after a diagnosis of severe dysplasia (P < .001). The annual incidence of gastric cancer was 0.1% for patients with atrophic gastritis, 0.25% for intestinal metaplasia, 0.6% for mild-to-moderate dysplasia, and 6% for severe dysplasia within 5 years after diagnosis. Risk factors for gastric cancer development were increasing severity of premalignant gastric lesions at initial diagnosis (eg, severe dysplasia, hazard ratio 40.14, 95% confidence interval 32.2-50.1), increased age (eg, 75-84 years, hazard ratio 3.75, 95% confidence interval 2.8-5.1), and male gender (hazard ratio 1.50, 95% CI 1.3-1.7). CONCLUSIONS: Patients with premalignant gastric lesions are at considerable risk of gastric cancer. As current surveillance of these patients is inconsistent with their cancer risk, development of guidelines is indicated. PMID- 18395076 TI - Parental obesity and offspring serum alanine and aspartate aminotransferase levels: the Framingham heart study. AB - BACKGROUND & AIMS: Obesity is an important correlate of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. We sought to examine the relations between parental obesity and the serum ALT and AST levels among offspring in a community-based sample. METHODS: Participants (n = 1732) of the Framingham Offspring Study (50% women; mean age, 42 years) who had serum ALT and AST measurements and both parents in the original Framingham cohort were studied. Study participants were grouped into early-onset parental obesity (n = 193) (at least one parent obese), later-onset parental obesity (n = 460), and no parental obesity (n = 1079) subgroups. The association between elevated ALT or AST levels and parental obesity was tested using generalized estimating equations to account for familial correlations. RESULTS: In multivariable analysis including adjustment for offspring obesity, significantly higher ALT levels were observed among individuals with paternal early-onset obesity as compared with those without paternal obesity (P = .02). Offspring with early-onset paternal obesity were more likely to have elevated ALT levels compared with those without paternal obesity (odds ratio, 1.75; 95% confidence interval, 1.06-2.89; P = .03). There was no association with elevated ALT levels among offspring with maternal early-onset obesity (odds ratio, 1.10; 95% confidence interval, 0.76-1.59; P = .61). There was no association between parental obesity and serum AST levels. CONCLUSIONS: Early-onset paternal obesity, but not maternal obesity, increases the odds of elevated serum ALT levels in offspring, suggesting a predisposition to developing elevated serum ALT levels that may be mediated through familial early-onset obesity. PMID- 18395077 TI - Performance of transient elastography for the staging of liver fibrosis: a meta analysis. AB - BACKGROUND & AIMS: Transient elastography has been studied in a multitude of liver diseases for the staging of liver fibrosis with variable results. A meta analysis was performed to assess the overall performance of transient elastography for the diagnosis of liver fibrosis and to analyze factors influencing the diagnostic accuracy. METHODS: Literature databases and international conference abstracts were searched. Inclusion criteria were as follows: evaluation of transient elastography, liver biopsy as reference, and assessment of the area under the receiver operating characteristic curve (AUROC). The meta-analysis was performed using the random-effects model for the AUROC, summary receiver operating curve techniques, as well as meta-regression approaches. RESULTS: Fifty studies were included in the analysis. The mean AUROC for the diagnosis of significant fibrosis, severe fibrosis, and cirrhosis were 0.84 (95% confidence interval [CI], 0.82-0.86), 0.89 (95% CI, 0.88-0.91), and 0.94 (95% CI, 0.93-0.95), respectively. For the diagnosis of significant fibrosis a significant reduction of heterogeneity of the AUROC was found when differentiating between the underlying liver diseases (P < .001). Other factors influencing the AUROC were the scoring system used and the country in which the study was performed. Age, body mass index, and biopsy quality did not have a significant effect on the AUROC. CONCLUSIONS: Transient elastography can be performed with excellent diagnostic accuracy and independent of the underlying liver disease for the diagnosis of cirrhosis. However, for the diagnosis of significant fibrosis, a high variation of the AUROC was found that is dependent on the underlying liver disease. PMID- 18395078 TI - The natural history of small-duct primary sclerosing cholangitis. AB - BACKGROUND & AIMS: The long-term prognosis of patients with small-duct primary sclerosing cholangitis (PSC) remains incompletely characterized. We aimed at determining the natural history and long-term outcomes of a large number of patients with small-duct PSC. METHODS: Data from 83 patients with well characterized small-duct PSC from several medical institutions in Europe and the United States were combined. Each patient with small-duct PSC was randomly matched to 2 patients with large-duct PSC by age, gender, calendar year of diagnosis, and institution. RESULTS: The median age at diagnosis in both groups was 38 years (61% males). Nineteen (22.9%) of the 83 patients with small-duct PSC progressed to large-duct PSC in a median of 7.4 (interquartile range [IQR], 5.1 14) years. One patient with small-duct PSC who progressed to large-duct PSC was diagnosed with cholangiocarcinoma but after progression to large-duct PSC; 20 patients with large-duct PSC developed cholangiocarcinoma. Patients with small duct PSC had a significantly longer transplantation-free survival compared with large-duct PSC patients (13 years [IQR, 10-17] vs 10 years [IQR, 6-14], respectively; hazard ratio, 3.04; 95% confidence interval: 1.82-5.06; P < .0001). Two patients with small-duct PSC who underwent liver transplantation had recurrence of small-duct PSC in the graft 9 and 13 years, respectively, after transplantation. CONCLUSIONS: Small-duct PSC is a disease of progressive potential but associated with a better long-term prognosis as compared with large duct PSC. Small-duct PSC may recur after liver transplantation. Cholangiocarcinoma does not seem to occur in patients with small-duct PSC, unless the disease has progressed to large-duct PSC. PMID- 18395079 TI - Prevalence and clinical profile of pancreatic cancer-associated diabetes mellitus. AB - BACKGROUND & AIMS: Information on the clinical profile of pancreatic cancer (PaC) associated diabetes (DM) is limited. We compared the prevalence and clinical characteristics of DM in subjects with and without PaC. METHODS: We prospectively recruited 512 newly diagnosed PaC cases and 933 controls of similar age, who completed demographic and clinical questionnaires and had fasting blood glucose (FBG) levels measured at recruitment and after pancreaticoduodenectomy (n = 105). Subjects with a FBG level >126 mg/dL or who were on antidiabetic treatment were classified as having DM. RESULTS: DM was more prevalent (47% vs 7%; P < .001) and predominantly of new onset (<2-year duration) (74% vs 53%; P = .002) among cases compared with controls. Among PaC cases, those with DM (n = 243) were older (68 +/- 10 vs 64 +/- 12 years; P < .001), reported higher usual adult body mass index (30 +/- 6 vs 27 +/- 5 kg/m(2); P < .001), and had a greater frequency of family history of DM (47% vs 31%; P < .001) compared with those without DM (n = 269). After pancreaticoduodenectomy, while DM resolved in 17 of 30 patients (57%) with new-onset DM, its prevalence was unchanged in patients with long-standing DM (n = 11) (P = .009). CONCLUSIONS: PaC is a powerful diabetogenic state; DM associated with PaC is often new-onset, resolves following cancer resection, and appears to be associated with conventional risk factors for DM. New-onset DM in patients with PaC is likely induced by the tumor. PMID- 18395080 TI - Immune response against frameshift-induced neopeptides in HNPCC patients and healthy HNPCC mutation carriers. AB - BACKGROUND & AIMS: Colorectal cancers (CRC) associated with the hereditary nonpolyposis colorectal cancer (HNPCC) syndrome display high-level microsatellite instability (MSI-H) as a consequence of mismatch repair deficiency. HNPCC associated CRC frequently show features of a pronounced immune response, most likely resulting from the MSI-induced generation of novel tumor-specific carboxy terminal frameshift peptides (FSPs). However, the role of FSP-specific immune surveillance mechanisms in HNPCC are unknown at present. METHODS: The efficacy of tumor-infiltrating T cells isolated from MSI-H CRCs (n = 3) was examined by in vitro killing assays. FSP-specific T-cell responses were measured by enzyme linked immunospot in the peripheral blood from patients with MSI-H CRC (n = 32), healthy HNPCC mutation carriers (n = 16), patients with microsatellite stable (MSS) CRC (n = 17), and healthy donors (n = 22). RESULTS: Tumor-infiltrating T cells isolated from MSI-H CRCs specifically recognized MSI-induced FSPs and showed cytotoxic activity against MSI-H but not MSS CRC cells. FSP-specific T cell responses were detected in the majority of peripheral blood samples from patients with MSI-H but not MSS CRC. Interestingly, FSP-specific T-cell reactivity was already detectable in the peripheral blood of healthy HNPCC family members with germline mutations but without history of tumor development. CONCLUSIONS: These data suggest that FSPs presented by DNA mismatch repair deficient CRC cells are effectively recognized by the patient's immune system and may explain the characteristic clinicopathologic features of HNPCC-associated but also sporadic MSI-H CRCs. These observations are of high relevance for the development of FSP-based vaccination approaches, particularly for the preventive application in HNPCC mutation carriers. PMID- 18395081 TI - The specific immune response to tumor antigen CP1 and its correlation with improved survival in colon cancer patients. AB - BACKGROUND & AIMS: The present study was undertaken to determine the expression of a newly identified tumor antigen cancer-placenta 1 (CP1) in colorectal carcinoma (CRC) and explore the CP1-specific immune response in CRC patients and its correlation with patient survival. METHODS: CP1 expression was determined by reverse-transcription polymerase chain reaction, immunohistochemistry, and Western blot analysis. Serum antibodies against CP1 were detected by enzyme linked immunosorbent assay, and T-cell response was measured by interferon gamma/granzyme-B release enzyme-linked immunospot assays. The HLA-A2-restricted epitopes in CP1 were predicted by bioinformatics and then experimentally validated by enzyme-linked immunospot assay. RESULTS: CP1 expression was detected in a significant number of CRC tissues, reaching 47.6% at the messenger RNA (mRNA) level and 28.6% at the protein level. Of patients with CP1 mRNA(+) tumors, more than 50% had CP1-responsive CD4(+) and CD8(+) T cells and 30% spontaneous occurring antibodies against CP1. Further studies revealed 2 dominant HLA-A2 restricted epitopes in the CP1 antigen: p31-39 and p58-66. In a follow-up study up to 33 months after surgery, 9 of the 10 patients with CP1-specific CD8 T-cell response survived, whereas 6 of the 8 nonresponders died. Kaplan-Meier analysis indicated a significant correlation between T-cell response and patient survival. CONCLUSIONS: CP1 represents a new class of tumor-specific shared antigen. Its high expression in CRC tissues, prevalence of CP1-specific immune responses in CP1 mRNA(+) CRC patients, and positive correlation with survival suggest that the antigen may be a useful target for cancer immunotherapy. PMID- 18395082 TI - Kruppel-like factor 5 mediates transmissible murine colonic hyperplasia caused by Citrobacter rodentium infection. AB - BACKGROUND & AIMS: Kruppel-like factor 5 (KLF5) is a transcription factor that is highly expressed in proliferating crypt cells of the intestinal epithelium. KLF5 has a pro-proliferative effect in vitro and is induced by mitogenic and stress stimuli. To determine whether KLF5 is involved in mediating proliferative responses to intestinal stressors in vivo, we examined its function in a mouse model of transmissible murine colonic hyperplasia triggered by colonization of the mouse colon by the bacteria Citrobacter rodentium. METHODS: Heterozygous Klf5 knockout (Klf5(+/-)) mice were generated from embryonic stem cells carrying an insertional disruption of the Klf5 gene. Klf5(+/-) mice or wild-type (WT) littermates were infected with C rodentium by oral gavage. At various time points postinfection, mice were killed and distal colons were harvested. Colonic crypt heights were determined morphometrically from sections stained with H&E. Frozen tissues were stained by immunofluorescence using antibodies against Klf5 and the proliferation marker, Ki67, to determine Klf5 expression and numbers of proliferating cells per crypt. RESULTS: Infection of WT mice with C rodentium resulted in a 2-fold increase in colonic crypt heights at 14 days postinfection and was accompanied by a 1.7-fold increase in Klf5 expression. Infection of Klf5(+/-) mice showed an attenuated induction of Klf5 expression, and hyperproliferative responses to C rodentium were reduced in the Klf5(+/-) animals as compared with WT littermates. CONCLUSION: Our study shows that Klf5 is a key mediator of crypt cell proliferation in the colon in response to pathogenic bacterial infection. PMID- 18395083 TI - Gliadin activates HLA class I-restricted CD8+ T cells in celiac disease intestinal mucosa and induces the enterocyte apoptosis. AB - BACKGROUND & AIMS: The extensive infiltration of CD8(+) T cells in the intestinal mucosa of celiac disease (CD) patients is a hallmark of the disease. We identified a gliadin peptide (pA2) that is selectively recognized by CD8(+) T cells infiltrating intestinal mucosa of HLA-A2(+) CD patients. Herein, we investigated the phenotype, the tissue localization, and the effector mechanism of cells responsive to pA2 by using the organ culture of CD intestinal mucosa. The target of pA2-mediated cytotoxicity was also investigated by using the intestinal epithelial cell lines Caco2 and HT29, A2(+) and A2(-), respectively, as target cells. METHODS: Jejunal biopsy specimens from CD patients were cultured in vitro with pA2, and cellular activation was evaluated by immunohistochemistry and cytofluorimetric analysis. Cytotoxicity of pA2-specific, intestinal CD8(+) T cells was assayed by granzyme-B and interferon-gamma release and by apoptosis of target cells. RESULTS: pA2 challenge of A2(+) CD mucosa increased the percentage of CD8(+)CD25(+) and of CD80(+) cells in the lamina propria, the former mainly localized beneath the epithelium, as well as the number of terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling-positive cells (TUNEL(+)) in the epithelium. Intraepithelial CD3(+) cells and enterocyte expression of Fas were also increased. CD8(+)CD25(+) and CD8(+)FASL(+) T cells were significantly increased in cell preparations from biopsy specimens cultured with pA2. CD8(+) T-cell lines released both granzyme-B and interferon-gamma following recognition of pA2 when presented by Caco2 and not by HT29. CONCLUSIONS: These data indicate that gliadins contain peptides able to activate, through a TCR/HLA class I interaction, CD8-mediated response in intestinal CD mucosa and to induce the enterocyte apoptosis. PMID- 18395084 TI - Direct presentation of antigen by lymph node stromal cells protects against CD8 T cell-mediated intestinal autoimmunity. AB - BACKGROUND & AIMS: Disruption of the enteric glial cell (EGC) network is an early pathologic feature in Crohn's disease. To determine the contribution of antigen specific CD8 and CD4 T cells to the breakdown of the EGC network, we studied specific autoimmune targeting of an ectopic antigen expressed by EGCs. METHODS: Transgenic mice (GFAP-HA), which express the influenza hemagglutinin (HA) in EGCs, were either crossed with mice transgenic for a T-cell receptor (TCR) specific for a major histocompatibility complex (MHC) class I epitope of HA (CL4 TCR) or were adoptively transferred with conventional CL4 T cells. These were compared with GFAP-HA mice transferred with conventional T cells specific for an MHC class II epitope of HA (6.5). RESULTS: Both CD8 and CD4 T-cell subtypes were activated in vivo in an antigen-specific manner; however, they differed substantially in their ability to expand in the mesenteric lymph nodes, trigger proinflammatory cytokines, and induce autoimmune damage in the intestine. Direct presentation of antigen, provided by lymph node stromal cells, caused the activation and deletion of CD8 T cells. This mechanism of T-cell tolerance did not affect CD4 T cells, which produced antigen-specific lethal autoimmunity. CONCLUSIONS: Our observations support a recently identified mechanism of peripheral T-cell tolerance that specifically protects against autoimmunity mediated by conventional CD8 T cells. Furthermore, we show that conventional CD4 T cells are not affected by this mechanism of tolerance, and their targeting of EGCs produces lethal intestinal autoimmunity. PMID- 18395085 TI - Regulation of gut inflammation and th17 cell response by interleukin-21. AB - BACKGROUND & AIMS: Interleukin (IL)-21, a T-cell-derived cytokine, is overproduced in inflammatory bowel diseases (IBD), but its role in the pathogenesis of gut inflammation remains unknown. We here examined whether IL-21 is necessary for the initiation and progress of experimental colitis and whether it regulates specific pathways of inflammation. METHODS: Both dextran sulfate sodium colitis and trinitrobenzene sulfonic acid-relapsing colitis were induced in wild-type and IL-21-deficient mice. CD4(+)CD25(-) T cells from wild-type and IL-21-deficient mice were differentiated in T helper cell (Th)17-polarizing conditions, with or without IL-21 or an antagonistic IL-21R/Fc. We also examined whether blockade of IL-21 by anti-IL-21 antibody reduced IL-17 in cultures of IBD lamina propria CD3(+) T lymphocytes. Cytokines were evaluated by real-time polymerase chain reaction and/or enzyme-linked immunosorbent assay. RESULTS: High IL-21 was seen in wild-type mice with dextran sulfate sodium- and trinitrobenzene sulfonic acid-relapsing colitis. IL-21-deficient mice were largely protected against both colitides and were unable to up-regulate Th17-associated molecules during gut inflammation, thus suggesting a role for IL-21 in controlling Th17 cell responses. Indeed, naive T cells from IL-21-deficient mice failed to differentiate into Th17 cells. Treatment of developing Th17 cells from wild-type mice with IL-21R/Fc reduced IL-17 production. Moreover, in the presence of transforming growth factor-beta1, exogenous IL-21 substituted for IL-6 in driving IL-17 induction. Neutralization of IL-21 reduced IL-17 secretion by IBD lamina propria lymphocytes. CONCLUSIONS: These results indicate that IL-21 is a critical regulator of inflammation and Th17 cell responses in the gut. PMID- 18395087 TI - Localization, trafficking, and significance for acid secretion of parietal cell Kir4.1 and KCNQ1 K+ channels. AB - BACKGROUND & AIMS: K(+) recycling at the apical membrane of gastric parietal cells is a prerequisite for gastric acid secretion. Two K(+) channels are currently being considered for this function, namely KCNQ1 and inwardly rectifying K(+) channels (Kir). This study addresses the subcellular localization, trafficking, and potential functional significance of KCNQ1 and Kir4.1 channels during stimulated acid secretion. METHODS: The effect of pharmacologic KCNQ1 blockade on acid secretion was studied in cultured rat and rabbit parietal cells and in isolated mouse gastric mucosa. The subcellular localization of KCNQ1 and Kir4.1 was determined in highly purified membrane fractions by Western blot analysis as well as in fixed and living cells by confocal microscopy. RESULTS: In cultured parietal cells and in isolated gastric mucosa, a robust acid secretory response was seen after complete pharmacologic blockade of KCNQ1. Both biochemical and morphologic data demonstrate that Kir4.1 and KCNQ1 colocalize with the H(+)/K(+)-ATPase but do so in different tubulovesicular pools. All Kir4.1 translocates to the apical membrane after stimulation in contrast to only a fraction of KCNQ1, which mostly remains cytoplasmic. CONCLUSIONS: Acid secretion can be stimulated after complete pharmacologic blockade of KCNQ1 activity, suggesting that additional apical K(+) channels regulate gastric acid secretion. The close association of Kir4.1 channels with H(+)/K(+)-ATPase in the resting and stimulated membrane suggests a possible role for Kir4.1 channels during the acid secretory cycle. PMID- 18395086 TI - Helicobacter pylori induces an antimicrobial response in rhesus macaques in a cag pathogenicity island-dependent manner. AB - BACKGROUND & AIMS: We used the rhesus macaque model to study the effects of the cag pathogenicity island (cag PAI) on the H pylori host-pathogen interaction. METHODS: H pylori-specific pathogen-free (SPF) monkeys were experimentally challenged with wild-type (WT) H pylori strain J166 (J166WT, n = 4) or its cag PAI isogenic knockout (J166Deltacag PAI, n = 4). Animals underwent endoscopy before and 1, 4, 8, and 13 weeks after challenge. Gastric biopsies were collected for quantitative culture, histopathology, and host gene expression analysis. RESULTS: Quantitative cultures showed that all experimentally challenged animals were infected with J166WT or its isogenic J166Deltacag PAI. Histopathology demonstrated that inflammation and expansion of the lamina propria were attenuated in animals infected with J166Deltacag PAI compared with J166WT. Microarray analysis showed that of the 119 up-regulated genes in the J166WT infected animals, several encode innate antimicrobial effector proteins, including elafin, siderocalin, DMBT1, DUOX2, and several novel paralogues of human-beta defensin-2. Quantitative RT-PCR confirmed that high-level induction of each of these genes depended on the presence of the cag PAI. Immunohistochemistry confirmed increased human-beta defensin-2 epithelial cell staining in animals challenged with J166WT compared with either J166Deltacag PAI-challenged or uninfected control animals. CONCLUSIONS: We propose that one function of the cag PAI is to induce an antimicrobial host response that may serve to increase the competitive advantage of H pylori in the gastric niche and could even provide a protective benefit to the host. PMID- 18395088 TI - Gastrin-mediated interleukin-8 and cyclooxygenase-2 gene expression: differential transcriptional and posttranscriptional mechanisms. AB - BACKGROUND & AIMS: Gastrin induces the expression of cyclooxygenase (COX)-2 and interleukin (IL)-8; however, the mechanism(s), especially in gastric epithelial cells, is not well understood. Here, we have determined the intracellular mechanisms mediating gastrin-dependent gene expression. METHODS: AGS-E human gastric cancer cell line stably expressing cholecystokinin-2 receptor was treated with amidated gastrin-17. Real-time polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay were performed to determine COX-2 and IL-8 expression and Akt, Erk, and p38 phosphorylation. Gene promoter activity was determined by luciferase assay. Electrophoretic mobility shift assay analysis was performed for nuclear factor kappaB (NF-kappaB) and activator protein-1 activity. RNA stability was determined after actinomycin D treatment. HuR localization was determined by immunocytochemistry. RESULTS: Gastrin induced COX-2 and IL-8 expression in AGS-E cells, which was inhibited by phosphatidylinositol 3' kinase (PI3K) and p38 inhibitors. Gastrin-mediated Akt activation was observed to be downstream of p38. IL-8 expression was dependent on COX-2-mediated prostaglandin E(2) synthesis. In the presence of an NF-kappaB inhibitor MG132, IL-8 transcription was inhibited, but not that of COX-2. This was confirmed after knockdown of the p65 RelA subunit of NF-kappaB. Further studies showed that COX-2 gene transcription is regulated by activator protein-1. Gastrin increased the stability of both COX-2 and IL-8 messenger RNA (mRNA) in a p38-dependent manner, the half-life increasing from 31 minutes to 8 hours and approximately 4 hours, respectively. Gastrin, through p38 activity, also enhanced HuR expression, nucleocytoplasmic translocation, and enhanced COX-2 mRNA binding. CONCLUSIONS: Gastrin differentially induces COX-2 and IL-8 expression at the transcriptional and posttranscriptional levels by PI3K and p38 mitogen-activated protein kinase pathways, respectively. PMID- 18395089 TI - Progenitors of interstitial cells of cajal in the postnatal murine stomach. AB - BACKGROUND & AIMS: Maintaining the integrity of networks of interstitial cells of Cajal (ICC) is essential to preserve orderly contractile activity and neuroregulation in the gastrointestinal tract and to restore these functions after tissue damage or surgeries. Maintenance of ICC requires insulin-dependent or insulin-like growth factor I (IGF-I)-dependent production of membrane-bound stem cell factor (SCF) and may involve regeneration from local progenitors. Our goal was to identify ICC precursors in postnatal murine gastric muscles. METHODS: We used flow cytometry and immunohistochemistry to examine freshly dissected and cultured muscles for cells expressing CD34, an adhesion molecule expressed by stromal tumors; CD44, which occurs on mesenchymal stem cells; and receptors for SCF (Kit), insulin (Insr), and IGF-I (Igf1r). Slow waves were studied by intracellular recording. RESULTS: In gastric muscles, we identified rare, Kit(low)CD44(+)CD34(+)Insr(+)Igf1r(+) cells resembling common embryonic precursors of ICC and smooth muscle. These putative progenitors were absent from organotypic cultures lacking mature ICC (Kit(+)CD44(+)CD34(-)Insr(-)Igf1r(-)) due to prolonged insulin/IGF-I deprivation but were rescued by IGF-I that also prevented ICC loss. Soluble SCF failed to prevent the loss of mature ICC but dramatically expanded the putative progenitors, which supported robust slow wave activity despite retaining an immature, Kit(+)CD44(+)CD34(+)Insr(+)Igf1r(+) phenotype. Differentiation of these cells into mature, network-forming ICC required IGF-I. Conversely, restoration of ICC networks by IGF-I after prolonged insulin and IGF-I deprivation required the survival of the presumed progenitors. CONCLUSIONS: Kit(low)CD44(+)CD34(+)Insr(+)Igf1r(+) cells may be local progenitors for gastric ICC and stromal tumors. Loss of these cells may contribute to gastrointestinal dysmotilities. PMID- 18395090 TI - Activation of extracellular signal-regulated protein kinase in sensory neurons after noxious gastric distention and its involvement in acute visceral pain in rats. AB - BACKGROUND & AIMS: Changes in the properties of visceral sensory neurons contribute to the development of gastrointestinal pain. However, little is known about the molecules involved in mechanosensation from the gastrointestinal tract. We investigated the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), a member of the mitogen-activated protein kinase cascade, in dorsal root ganglion (DRG) and nodose ganglion (NG) neurons by noxious gastric distention (GD) and its involvement in acute visceral pain in rats. METHODS: Electromyographic responses to gastric balloon distention through gastrostomy were recorded from the acromiotrapezius muscle in rats after splanchnic nerve resection or vagotomy and in control rats. We then examined the phosphorylated ERK1/2 (p-ERK1/2) labeling in the DRG and NG after GD using immunohistochemistry. RESULTS: Gastric distention induced p-ERK1/2 in DRG and NG neurons with a peak at 2 minutes after stimulation. We found a stimulus intensity-dependent increase in the number of activated neurons, and this activation corresponded well with the incidence of the visceromotor response. Most of these p-ERK1/2-labeled neurons were small- and medium-sized neurons that coexpressed transient receptor potential vanilloid 1 ion channel and acid-sensing ion channel 3. Splanchnic nerve resection, but not vagotomy, affected the visceromotor response, and attenuated the ERK1/2 activation in DRG neurons produced by GD. Furthermore, intrathecal administration of the mitogen-activated protein kinase kinase 1/2 inhibitor, U0126, altered the response to noxious GD. CONCLUSIONS: The activation of ERK1/2 pathways in DRG neurons by noxious GD may be correlated with functional activity, and may be involved in acute visceral pain. PMID- 18395091 TI - Perturbation of hoxb5 signaling in vagal neural crests down-regulates ret leading to intestinal hypoganglionosis in mice. AB - BACKGROUND & AIMS: The enteric nervous system (ENS) controls intestinal peristalsis, and defective development of this system results in hypo/aganglionosis, as seen in Hirschsprung's disease. In the embryo, vagal neural crest cells (NCC) migrate and colonize the intestine rostrocaudally then differentiate into the ganglia of the ENS. Vagal NCC express the homeobox gene Hoxb5, a transcriptional activator, in human and mouse, so we used transgenic mice to investigate the function of Hoxb5 and the receptor tyrosine kinase gene Ret, which is affected in many patients with Hirschsprung's disease, in ENS development. METHODS: We perturbed the Hoxb5 pathway by expressing a chimeric protein enb5, in which the transcription activation domain of Hoxb5 was replaced with the repressor domain of the Drosophila engrailed protein (en), in vagal NCC. This enb5 transcriptional repressor competes with wild-type Hoxb5 for binding to target genes, exerting a dominant negative effect. RESULTS: We observed that 30.6% +/- 2.3% of NCC expressed enb5 and that these enb5-expressing NCC failed to migrate to the distal intestine. A 34%-37% reduction of ganglia (hypoganglionosis) and slow peristalsis and, occasionally, absence of ganglia and intestinal obstruction were observed in enb5-expressing mice. Ret expression was markedly reduced or absent in NCC and ganglia, and enb5 blocked Hoxb5 induction of Ret in neuroblastoma cells. CONCLUSIONS: Our data indicate that Ret is a downstream target of Hoxb5 whose perturbation causes Ret haploinsufficiency, impaired NCC migration, and hypo/aganglionosis, suggesting that Hoxb5 may contribute to the etiology of Hirschsprung's disease. PMID- 18395092 TI - Dramatically increased intestinal absorption of cholesterol following hypophysectomy is normalized by thyroid hormone. AB - BACKGROUND & AIMS: Hypopituitarism is associated with dyslipidemia, and feeding hypophysectomized rats cholesterol induces severe hypercholesterolemia. This study aimed to unravel further how hypophysectomy alters cholesterol and bile acid metabolism. METHODS: Intact and hypophysectomized rats were studied during challenge with dietary cholesterol and ezetimibe and upon hormonal substitution with growth hormone, cortisone, and thyroid hormone. RESULTS: Five findings were established in hypophysectomized rats: (1) The intestinal absorption of cholesterol is doubled. (2) Treatment with ezetimibe abolishes the increases in serum and liver cholesterol. (3) Only thyroid hormone treatment normalizes the increased absorption of cholesterol. (4) The intestinal gene expression of cholesterol transporters NPC1L1 and ABCG5/G8 is unaltered, whereas the hepatic expression of ABCG5/G8 is diminished but strongly stimulated by thyroid hormone. The latter mechanism was supported by measurements of biliary cholesterol and of fecal neutral steroids. (5) The reduced hepatic expression of ABCG5/G8 and Cyp7a1 was normalized by cholesterol feeding, suggesting that other nonestablished mechanisms under pituitary control are important to maintain rats resistant to dietary cholesterol. CONCLUSIONS: The intestinal absorption of dietary cholesterol is under pituitary control largely exerted by thyroid hormone. Hepatic secretion of cholesterol and ABCG5/G8 expression are strongly stimulated in hypophysectomized rats during treatment with thyroid hormone. PMID- 18395093 TI - Cell type-dependent pro- and anti-inflammatory role of signal transducer and activator of transcription 3 in alcoholic liver injury. AB - BACKGROUND & AIMS: Signal transducer and activator of transcription 3 (STAT3) is known to be activated in human alcoholic liver disease, but its role in the pathogenesis of alcoholic liver injury remains obscure. METHODS: The role of STAT3 in alcoholic liver injury was investigated in hepatocyte-specific STAT3 knockout (H-STAT3KO) mice and macrophage/neutrophil-specific STAT3 KO (M/N STAT3KO) mice. Alcoholic liver injury was achieved by feeding mice a liquid diet containing 5% ethanol for up to 8 weeks. RESULTS: Compared with wild-type mice, feeding H-STAT3KO mice with an ethanol-containing diet induced greater hepatic steatosis, hypertriglyceridemia, and hepatic expression of lipogenic genes (sterol regulatory element-binding protein, fatty acid synthase, acetyl-CoA carboxylase-1, and stearoyl-CoA desaturase 1), but less inflammation and lower expression of hepatic proinflammatory cytokines. In contrast, ethanol-fed M/N STAT3KO mice showed more hepatic inflammation, worse injury, and increased hepatic expression of proinflammatory cytokines compared with wild-type mice. Kupffer cells isolated from ethanol-fed H-STAT3KO mice produced similar amounts of reactive oxygen species and tumor necrosis factor alpha, whereas Kupffer cells from M/N-STAT3KO mice produced more reactive oxygen species and tumor necrosis factor alpha compared with wild-type controls. CONCLUSIONS: These findings suggest that STAT3 regulates hepatic inflammation in a cell type-dependent manner during alcoholic liver injury: STAT3 in hepatocytes promotes whereas STAT3 in macrophages/Kupffer cells suppresses inflammation. In addition, activation of hepatocellular STAT3 ameliorates alcoholic fatty liver via inhibition of sterol regulatory element-binding protein 1c expression. PMID- 18395094 TI - Nuclear factor-eythroid 2-related factor 2 prevents alcohol-induced fulminant liver injury. AB - BACKGROUND & AIMS: The transcription factor nuclear factor-eythroid 2-related factor 2 (Nrf2(-/-)) is essential for protecting cells against xenobiotic and oxidative stress. Increased oxidative stress has been implicated in the pathophysiology of many diseases including ethanol-induced liver disease. Therefore, the role of Nrf2(-/-) in ethanol-induced liver injury was investigated. METHODS: Wild-type and Nrf2(-/-) mice were fed with the ethanol diet, followed by examination of liver pathology, mortality, and ethanol metabolism. RESULTS: Nrf2(-/-) mice displayed a dramatically increased mortality associated with liver failure when fed doses of ethanol that were tolerated by WT mice. Nrf2(-/-) mice showed a significantly reduced ability to detoxify acetaldehyde, leading to an accumulation of the toxic metabolite. Loss of Nrf2(-/ ) caused a marked steatosis in livers of ethanol-fed mice, and Srebp1 was identified as a candidate transcription factor responsible for lipogenic enzyme induction. Furthermore, ethanol consumption led to a progressive depletion of total and mitochondrial reduced glutathione, which was associated with more pronounced structural and functional changes to mitochondria of Nrf2(-/-) mice. In addition, ethanol feeding elicited an aggravated inflammatory response mediated by Kupffer cells in Nrf2(-/-) mice as shown by an increased tumor necrosis factor-alpha secretion and activation of the interleukin-6/Stat-3 pathway. Together these changes lead to a vicious cycle of accumulating hepatocellular damage, ultimately leading to liver failure and death of Nrf2(-/-) mice. CONCLUSIONS: Our data establish a central role for Nrf2(-/-) in the protection against ethanol-induced liver injury. PMID- 18395095 TI - Keratin mutation predisposes to mouse liver fibrosis and unmasks differential effects of the carbon tetrachloride and thioacetamide models. AB - BACKGROUND & AIMS: Keratins 8 and 18 (K8/K18) are important hepatoprotective proteins. Animals expressing K8/K18 mutants show a marked susceptibility to acute/subacute liver injury. K8/K18 variants predispose to human end-stage liver disease and associate with fibrosis progression during chronic hepatitis C infection. We sought direct evidence for a keratin mutation-related predisposition to liver fibrosis using transgenic mouse models because the relationship between keratin mutations and cirrhosis is based primarily on human association studies. METHODS: Mouse hepatofibrosis was induced by carbon tetrachloride (CCl(4)) or thioacetamide. Nontransgenic mice, or mice that over express either human Arg89-to-Cys (R89C mice) or wild-type K18 (WT mice) were used. The extent of fibrosis was evaluated by quantitative real-time reverse transcription polymerase chain reaction of fibrosis-related genes, liver hydroxyproline measurement, and Picro-Sirius red staining and collagen immunofluorescence staining. RESULTS: Compared with control animals, CCl(4) led to similar liver fibrosis but increased injury in K18 R89C mice. In contrast, thioacetamide caused more severe liver injury and fibrosis in K18 R89C as compared with WT and nontransgenic mice and resulted in increased messenger RNA levels of collagen, tissue inhibitor of metalloproteinase 1, matrix metalloproteinase 2, and matrix metalloproteinase 13. Analysis in nontransgenic mice showed that thioacetamide and CCl(4) have dramatically different molecular expression responses involving cytoskeletal and chaperone proteins. CONCLUSIONS: Over expression of K18 R89C predisposes transgenic mice to thioacetamide- but not CCl(4)-induced liver fibrosis. Differences in the keratin mutation-associated fibrosis response among the 2 models raise the hypothesis that keratin variants may preferentially predispose to fibrosis in unique human liver diseases. Findings herein highlight distinct differences in the 2 widely used fibrosis models. PMID- 18395096 TI - Hepatocyte growth factor suppresses profibrogenic signal transduction via nuclear export of Smad3 with galectin-7. AB - BACKGROUND & AIMS: Hepatocyte growth factor (HGF) and transforming growth factor beta (TGF-beta) regulate diversified cellular functions and often act antagonistically against each other. For example, TGF-beta is the most potent factor accelerating liver fibrosis, whereas HGF treatment prevents its progression. Here, we propose a novel molecular mechanism by which HGF counter represses TGF-beta-stimulated profibrogenic signal transduction. METHODS: Effects of HGF on TGF-beta-responsive gene transcription of type I collagen, the major matrix component of fibrotic liver, were examined by using cultured hepatic stellate cells (HSC) and transgenic mice harboring alpha2(I) collagen gene (COL1A2) promoter. Expression and subcellular localization of Smad3 were determined by Western blot analyses and immunofluorescence staining, respectively. A mass spectrometric analysis was employed to identify immunoprecipitated proteins with antiphospho-Smad2/3 antibodies. RESULTS: Over expression of HGF inhibited COL1A2 transcription in cultured HSC and suppressed activation of COL1A2 promoter in liver tissue induced by carbon tetrachloride administration. A mass spectrometric analysis identified galectin-7 as one of the immunoprecipitated proteins with antiphospho-Smad2/3 antibodies following HGF treatment. HGF accelerated nuclear export of Smad3 by enhancing its interaction with galectin-7. Transfection of cells with galectin-7 small interfering RNA inhibited nuclear export of Smad3 and abolished suppressive effect of HGF on expression of TGF-beta-responsive genes such as COL1A2 and plasminogen activator inhibitor-1. On the other hand, over expression of galectin-7 suppressed TGF-beta stimulated expression of those target genes. CONCLUSIONS: These results reveal a novel function of intracellular galectin-7 as a transcriptional regulator via its interaction with Smad3 and provide a molecular basis for the antifibrotic effect of HGF. PMID- 18395097 TI - EPS15R, TASP1, and PRPF3 are novel disease candidate genes targeted by HNF4alpha splice variants in hepatocellular carcinomas. AB - BACKGROUND & AIMS: The orphan nuclear receptor HNF4alpha is a member of the hepatic transcription factor network. This protein plays a pivotal role in liver development and hepatocellular differentiation. Nine splice variants have been identified, some of which are specifically regulated in disease. The role of HNF4alpha splice variants in hepatocellular carcinomas (HCC) is unknown. Here, we report an identification of novel candidate genes targeted by splice variants of HNF4alpha. METHODS: We used chromatin immunoprecipitation followed by cloning and sequencing of DNA. Expression of HNF4alpha P1 and P2 promoter-driven isoforms and of genes targeted by HNF4alpha were analyzed by quantitative reverse transcription polymerase chain reaction, Western blotting, electrophoretic mobility shift assay, and immunohistochemistry. RESULTS: We observed a remarkable switch in gene and protein expression from P1 to P2 promoter-driven fetal isoforms of HNF4alpha in transgenic livers and HCCs of epidermal growth factor (EGF) overexpressing mice and in human HCCs. We further identified EGF-receptor substrate (EPS15R), related EPS15, the premessenger RNA processing factor 3 (PRPF3), and taspase 1 (TASP1) as novel HNF4alpha disease regulated genes with induced expression in mouse and in human HCCs. We suggest EPS15 and EPS15R mediated internalization of activated EGF receptor to result in receptor recycling as to reinforce the proliferative response to EGF. Regulation of the type 2 asparaginase TASP1 and of the splicing factor PRPF3 further documents a switch to fetal liver programs in HCC. CONCLUSIONS: We report induction of P2 promoter-driven HNF4alpha splice variants and regulation of disease candidate genes in EGF-induced mouse and human HCC. PMID- 18395098 TI - Severe bile salt export pump deficiency: 82 different ABCB11 mutations in 109 families. AB - BACKGROUND & AIMS: Patients with severe bile salt export pump (BSEP) deficiency present as infants with progressive cholestatic liver disease. We characterized mutations of ABCB11 (encoding BSEP) in such patients and correlated genotypes with residual protein detection and risk of malignancy. METHODS: Patients with intrahepatic cholestasis suggestive of BSEP deficiency were investigated by single-strand conformation polymorphism analysis and sequencing of ABCB11. Genotypes sorted by likely phenotypic severity were correlated with data on BSEP immunohistochemistry and clinical outcome. RESULTS: Eighty-two different mutations (52 novel) were identified in 109 families (9 nonsense mutations, 10 small insertions and deletions, 15 splice-site changes, 3 whole-gene deletions, 45 missense changes). In 7 families, only a single heterozygous mutation was identified despite complete sequence analysis. Thirty-two percent of mutations occurred in >1 family, with E297G and/or D482G present in 58% of European families (52/89). On immunohistochemical analysis (88 patients), 93% had abnormal or absent BSEP staining. Expression varied most for E297G and D482G, with some BSEP detected in 45% of patients (19/42) with these mutations. Hepatocellular carcinoma or cholangiocarcinoma developed in 15% of patients (19/128). Two protein-truncating mutations conferred particular risk; 38% (8/21) of such patients developed malignancy versus 10% (11/107) with potentially less severe genotypes (relative risk, 3.7 [confidence limits, 1.7-8.1; P = .003]). CONCLUSIONS: With this study, >100 ABCB11 mutations are now identified. Immunohistochemically detectable BSEP is typically absent, or much reduced, in severe disease. BSEP deficiency confers risk of hepatobiliary malignancy. Close surveillance of BSEP-deficient patients retaining their native liver, particularly those carrying 2 null mutations, is essential. PMID- 18395099 TI - ATP7B copper-regulated traffic and association with the tight junctions: copper excretion into the bile. AB - BACKGROUND & AIMS: The copper transporter ATP7B plays a central role in the elimination of excess copper by the liver into the bile, yet the site of its action remains controversial. The studies reported here examine the correspondence between the site of ATP7B action and distribution and the pathways of copper disposal by the liver. METHODS: Microscopy and cell fractionation studies of polarized Can 10 cells forming long-branched bile canaliculi have been used to study the cellular distribution of ATP7B. Copper excretion into the bile was studied in perfused rat liver. RESULTS: Copper excess provokes a massive download of the ATP7B retained in the trans-Golgi network into the bile canalicular membrane. Furthermore, a stable ATP7B pool is localized to the tight junctions that seal the bile canaliculi. The profile of Cu(64) excretion into the bile by isolated rat livers perfused under one-pass conditions provides evidence of copper excretion by 2 separate mechanisms, transcytosis across the hepatocyte and paracellular transport throughout the tight junctions. CONCLUSIONS: Whereas the ATP7B retained in the trans-Golgi-network is massively translocated to the bile canalicular membrane in response to increased copper levels, a pool of ATP7B associated with the tight junctions remains stable. In situ studies indicate that copper is excreted into the bile by 2 separate pathways. The results are discussed in the frame of the normal and impeded excretion of copper into the bile. PMID- 18395100 TI - Chemoprevention of colorectal neoplasia: the potential for personalized medicine. AB - CRC development is a multi-step process that spans 10 to 15 years, thereby providing an opportunity for early detection and even prevention. The poor survival rate of advanced CRC has prompted the emphasis on prevention of this disease. CRC screening and removal of adenomas is an effective intervention, and is the cornerstone of prevention. However, screening efforts have had limited impact due to less than optimal compliance with guidelines. Chemoprevention involves the long-term use of a variety of oral agents that can delay, prevent or even reverse the development of adenomas in the large bowel, thus interfering with the multi-step progessing from adenoma to carcinoma. This effect is of particular importance to individuals with a hereditary prediposition to colorectal neoplasia and to those who are especially susceptile to the environmental causes of CRC. NSAIDs have drawn the most attention as chemoprevention agents. Sulindac and celecoxib are effective in promoting poly regression in high risk individuals with Familial Adenomatous Polyposis (FAP). In the more common sporadic setting the APROVe (refecoxib), APC and PreSAP (Celecoxib) trials have shown a significant reduction in adenoma recurrence but important concerns exist regarding cardiovascular toxicity associated with selective COX-2 inhibitors. These landmark studies are very important, as they provide a proof of concept that we can prevent high risk adenomas that can lead to CRC development. The ideal chemopreventive agent remains to be discovered with great emphasis on need not to harm. Possibly, combinations of agents will maximize effectiveness while limiting drug toxicity. Finally, personalized approaches will include the ability to predict risk and toxicity. PMID- 18395101 TI - NSAIDs, risks, and gastroprotective strategies: current status and future. PMID- 18395102 TI - Immunology and the Lynch syndrome. PMID- 18395103 TI - Ultimately, it matters where, when, and what they'll meet. PMID- 18395104 TI - Progenitor cells of interstitial cells of Cajal: on the road to tissue repair. PMID- 18395105 TI - Could ATP7B export Cu(I) at the tight junctions and the apical membrane? PMID- 18395106 TI - Fear of loss, not promise of gain, drives practice patterns in Barrett's esophagus. PMID- 18395108 TI - Loperamide use for acute infectious diarrhea in children: safe and sound? PMID- 18395109 TI - The new look of colorectal cancer stem cells. PMID- 18395110 TI - vacA i-region subtyping. PMID- 18395111 TI - Insulin resistance and colorectal adenomas. PMID- 18395114 TI - Combination of hepatitis B viral factors and advanced liver disease in HBeAg negative patients: the more, the worse? PMID- 18395116 TI - HBV genotype and clinical outcome of chronic hepatitis B: facts and puzzles. PMID- 18395118 TI - The hominoid remains from the middle Miocene site of Pasalar, Turkey. Preface. PMID- 18395119 TI - A new hominoid species from the middle Miocene site of Pasalar, Turkey. AB - A new species of fossil hominoid is described from the middle Miocene deposits at Pasalar, Turkey. It is the less common of the two Pasalar species discussed by Martin and Andrews (1993), making up approximately 10% of the individuals in the Pasalar hominoid sample according to analyses of the minimum number of individuals. To the diagnostic features of I(1) described by Alpagut et al. (1990) and Martin and Andrews (1993) can now be added further diagnostic features of all the anterior teeth, as well as both upper premolars and P(3). These include discrete, nonmetric features and metric differences at all the noted tooth positions. Attempts to distinguish the upper and lower molars of the two species have so far been unsuccessful, with the possible exception of M(3). The morphology of the new species is similar in most respects to that of Kenyapithecus wickeri from Fort Ternan, especially concerning maxillary morphology. They share robust and moderately deep maxillary alveolar processes, a restricted maxillary sinus with an elevated and uncomplicated floor, lacking the compartmentalization evident to varying degrees in many other taxa, and a zygomatic process that originates and turns laterally fairly high above the alveolar margin. There are also a number of distinctive similarities in the dentition, particularly for I(1), C(1), P(4) and P(3). The I(1) morphology in particular, with greatly hypertrophied lingual marginal ridges bounding a uniformly thickened basal crown area, is distinctive among Miocene hominoids. All of these similarities serve to reinforce the differences noted by others between the derived morphology of K. wickeri and the more primitive morphology of Equatorius africanus from Maboko and Kipsaramon. The new species differs from K. wickeri in morphological details of most of the anterior and premolar teeth that are known for both species, despite the general morphological similarity, and in the size of I(1) versus I(2). One striking feature of the new species is a relatively large incisive fossa, although it cannot be determined if this is associated with an open palatine fenestra, as in many early Miocene hominoids, or a minimally overlapping palate and nasoalveolar clivus, as in some middle and late Miocene hominoids. PMID- 18395120 TI - Interproximal wear facets and tooth associations in the Pasalar hominoid sample. AB - Interproximal wear facets were examined on hominoid teeth from the middle Miocene site at Pasalar, Turkey. The aim was to find matches between adjacent premolar and molar teeth from single individuals that were collected in the field as isolated teeth and use them to reconstruct tooth rows. These were then used to investigate: (1) the wear gradient on the molar teeth; (2) the dispersal of teeth from single mandibles and maxillae; (3) the size ratios among the molars; and (4) the number of individuals represented by the hominoid sample. Facets were scored for size and shape and were assessed visually using photographs and superimposed outline drawings on acetate transparencies. Out of a sample of approximately 1,500 teeth collected between 1983 and 1996, 532 molars and 258 premolars produced apparent matches making up 160 tooth rows. These were then examined rigorously for morphological consistency and state of wear, and, employing the criterion that only the most unequivocal associations should be used, the final number was reduced to 48 tooth rows-31 mandibular and 17 maxillary. The tooth associations represent a minimum of 21 individuals and probably as many as 34. Molar wear was rapid, with M1s having almost twice as much wear as M3s, as measured by a wear-gradient index. The M2s are intermediate but generally closer to M1s in degree of wear, as are P4s. This wear pattern suggests either delayed eruption of M3s or extremely abrasive diets causing rapid, heavy wear. There is some indication that the wear patterns in Griphopithecus alpani and Kenyapithecus kizili are different, with the latter perhaps having a lower wear gradient, but the K. kizili sample is very small. In both species, the M2 is the largest molar and the M1 is the smallest. Separation of individual teeth in the 48 tooth associations varied from widely separated-up to 8.5m apart-to within a few centimeters of each other. One tooth row (D922) was found with the teeth in contact but the maxillary bone had dissolved away. Two dispersal mechanisms have been identified from earlier taphonomic work: transport of disarticulated elements to the fossil site and reworking of sediments by spring action. PMID- 18395121 TI - The deciduous dentition of Griphopithecus alpani from Pasalar, Turkey. AB - Seventy-four hominoid primary teeth have been recovered from the middle Miocene site of Pasalar, Turkey, constituting the largest sample of deciduous teeth for any species of fossil ape. Morphological features that characterize the permanent teeth of Griphopithecus alpani from the site have also been identified in some of these deciduous teeth, including a lingual pillar on the di(1)s. These features plus the overwhelming preponderance of G. alpani permanent teeth at the site suggest that all of the deciduous teeth belong to this species. Contrary to the situation in the permanent teeth, nothing in the morphology of the primary dentition suggests the representation of a second species. The age profile of the non-adult hominoids was reconstructed based on the degree and type of wear recorded on the dp4s, the most abundant deciduous tooth in the sample, assuming a similar eruption chronology to that of Pan troglodytes. This analysis indicates underrepresentation of very young individuals in the sample and high mortality for individuals belonging to the 3-5-years age cohort, a situation that could be due to the effects of stress related to weaning. The coefficient of variation and range-index values obtained for the majority of tooth types are equal to or greater than the comparable values in a sample of P. troglodytes, in some cases at much smaller sample sizes. One possible explanation for this is that there was greater sexual dimorphism in the G. alpani deciduous dentition than in Pan, which would mirror the condition of the permanent dentition. PMID- 18395122 TI - Hominoid phalanges from the middle Miocene site of Pasalar, Turkey. AB - Eleven proximal and ten intermediate partial or complete hominoid phalanges have been recovered from the middle Miocene site of Pasalar in Turkey. Based on species representation at Pasalar, it is likely that most or all of the phalanges belong to Griphopithecus alpani rather than Kenyapithecus kizili, but both species may be represented. All of the complete or nearly complete phalanges appear to be manual, so comparisons to extant and other fossil primate species were limited to manual phalanges. Comparisons were made to extant hominoid and cercopithecoid primate genera expressing a variety of positional repertoires and varying degrees of arboreality and terrestriality. The comparisons consisted of a series of bivariate indices derived from previous publications on Miocene catarrhine phalangeal morphology. The proximal phalanges have dorsally expanded proximal articular surfaces, which is characteristic of cercopithecoids and most other Miocene hominoids, and indicates that the predominant positional behaviors involved pronograde quadrupedalism. Among the extant primates, many of the proximal and intermediate phalangeal indices clearly distinguish more habitually terrestrial taxa from those that are predominantly arboreal, and especially from taxa that commonly engage in suspensory activities. For nearly every index, the values of the Pasalar phalanges occupy an intermediate position-most similar to values for Pan and, to a lesser extent, Macaca-indicating a generalized morphology and probably the use of both arboreal and terrestrial substrates. At least some terrestrial activity is also compatible with reconstructions of the Pasalar habitat. Most proximal and intermediate phalanges of other middle and late Miocene hominoids have similar index values to those of the Pasalar specimens, revealing broadly similar manual phalangeal morphology among many Miocene hominoids. PMID- 18395123 TI - Identification of a single birth cohort in Kenyapithecus kizili and the nature of sympatry between K. kizili and Griphopithecus alpani at Pasalar. AB - The sample of the less common hominoid species at Pasalar, Kenyapithecus kizili, is characterized by a number of unusual attributes. All ten of the upper central incisors attributed to this species show a distinct, identical pattern of two linear enamel hypoplasias. The two hypoplasias occur on the same portion of the labial crown face, revealing that the two hypoplasia-causing events occurred at the same stage of development in all individuals. The morphology of the two hypoplasias and the amount of time between them, as determined by both their separation and counts of perikymata, are also the same on all teeth. In addition, all of the approximately 70 teeth assigned to K. kizili appear to come from young adults based on degrees of wear; there are no younger or older individuals (diagnostic morphology at most tooth positions would be evident even with heavy wear). Thus, all of the K. kizili individuals (minimum number of individuals is nine: seven males, two females) appear to have died at essentially the same age. It is concluded that the most plausible interpretation of all these features is that the incisor hypoplasias were caused by the same two events in all the K. kizili individuals and that these individuals therefore represent a single birth cohort. As such, and because they died at essentially the same age, they would also have died at the same time, which is consistent with the catastrophic nature of the Pasalar deposits. The number of coincidences needed to explain all of the attributes of the K. kizili sample if these animals were born in, and died in, different years seems highly improbable. Moreover, the lack of a typical age class structure for the K. kizili sample, or any age-class structure at all beyond the one age class of young adult, strongly suggests that the species was not resident in the area that contributed to the Pasalar accumulation, and that K. kizili was not permanently sympatric with the other Pasalar hominoid, Griphopithecus alpani. Rather, the nine K. kizili individuals must have been transients in, or recent immigrants to, the area at the time of the events that led to the formation of the site. Recent observations on social associations in male chimpanzees offer at least a possible interpretive framework to explain this unprecedented occurrence in the primate fossil record. PMID- 18395125 TI - Submerged culture fermentation of "higher fungi": the macrofungi. PMID- 18395124 TI - A ferment of fermentations: reflections on the production of commodity chemicals using microorganisms. PMID- 18395126 TI - Bioprocessing using novel cell culture systems. PMID- 18395127 TI - Nanotechnology in the detection and control of microorganisms. PMID- 18395128 TI - Metabolic aspects of aerobic obligate methanotrophy. PMID- 18395129 TI - Bacterial efflux transport in biotechnology. PMID- 18395130 TI - Antibiotic resistance in the environment, with particular reference to MRSA. PMID- 18395131 TI - Host defense peptides in the oral cavity. PMID- 18395132 TI - Foreword. The 8th Annual European Round Table on Fabry Disease. PMID- 18395133 TI - Implications of early renal changes in Fabry disease. PMID- 18395134 TI - Newborn screening in Fabry disease: what can be achieved with early diagnosis? PMID- 18395135 TI - Prominence of glomerular and vascular changes in renal biopsies in children and adolescents with Fabry disease and microalbuminuria. PMID- 18395136 TI - Fabry disease--whom to treat and when. PMID- 18395137 TI - Early detection and treatment of Fabry disease. PMID- 18395138 TI - Early detection of Fabry disease: cardiac cases. PMID- 18395139 TI - Neurology in Fabry disease. PMID- 18395140 TI - The immunogenicity of therapeutic proteins and the Fabry antibody standardization initiative. PMID- 18395141 TI - Assessment of pain and somatosensory function in Fabry disease: early diagnosis. PMID- 18395142 TI - Optimising early warning scoring systems. PMID- 18395143 TI - Cancer Survivorship. Preface. PMID- 18395144 TI - Cancer survivors and survivorship research: a reflection on today's successes and tomorrow's challenges. AB - As advances in cancer medicine turn this once uniformly fatal illness into a curable disease for growing numbers and a chronic illness for many, understanding and meeting the needs of long-term cancer survivors and their caregivers has become a major public health challenge, a challenge made more urgent by the aging of the population. This article reviews the profile of today's cancer survivors along with the demographic information on what this profile might look like in the future. Current directions in and the knowledge gained from the growing body of cancer survivorship research and the science of the long-term and late consequences to individuals, families, and society of people living longer with a cancer history are delineated. PMID- 18395145 TI - Follow-up of adult cancer survivors: new paradigms for survivorship care planning. AB - The reality of cancer care in the twenty first century is that patients live longer and are more likely to receive care from multiple providers across diverse delivery systems over many years. To meet the challenge of optimal survivor care, a summary of cancer treatment and a formal plan for survivorship must be explained to patients and shared among providers at the end of cancer treatment. These plans must be dynamic documents that change with the circumstances of individual patients, the growth of knowledge, and the guidelines in specific relevant areas. In the new paradigm, open communication across the spectrum of survivor needs and concerns (as contained in a survivorship care plan) will successfully transition cancer patients to healthy survivors. Survivorship planning must become an integral part of every oncologist's education and practice. PMID- 18395146 TI - Survivors of childhood cancer: coming of age. AB - The development of curative therapy for most pediatric malignancies has produced a growing population of childhood cancer survivors who are at increased risk for a variety of health problems resulting from their cancer or its treatment. Risk based survivor care is recommended for all survivors. To optimize risk-based survivor care, several groups have organized health screening guidelines based on evidence from the literature linking specific therapeutic interventions with late treatment complications. This article addresses the scope of long-term health effects after pediatric cancer, the challenges in coordinating long-term survivor care, and health screening guideline resources available to facilitate survivor care. PMID- 18395148 TI - Testicular cancer patients: considerations in long-term follow-up. AB - Over the past 30 years, testicular tumors have become the paradigm for a curable adult cancer. Numerous factors have contributed to this success, including the introduction of newer treatment approaches, such as cisplatin-based combination chemotherapy and curative retroperitoneal lymph node dissection. Moreover, the last three decades have witnessed the evolution of newer diagnostic methods, improvements in staging, the evaluation of patient response, and the monitoring of relapse. These treatment successes have been accompanied by the emergence of the late effects of testicular cancer and its treatment, including second primary cancers, cardiovascular sequelae, the metabolic syndrome, gonadal toxicity, neurotoxicity, and pulmonary sequelae. An overview of these late effects and recommendations for patient follow-up are presented in this article. PMID- 18395147 TI - Hodgkin lymphoma: the follow-up of long-term survivors. AB - Hodgkin lymphoma (HL) is a disease that typically strikes children and young adults, with more than 80% of those affected being cured. Consequently, HL survivors can live for decades with the persistent and late-emerging effects of the disease and its treatment. The focus of this article is the long-term medical management of HL survivors, specifically those who are 5 years or more off therapy without evidence of relapse, when the focus of follow-up care shifts from detecting relapse to minimizing the morbidity associated with the late effects of treatment. PMID- 18395149 TI - The genetics of cancer survivorship. AB - Constitutional (hereditary) genetic variation and somatic genetic alterations acquired during transformation to the neoplastic phenotype are both critical determinants of cancer outcome, and can ultimately have a significant effect on cancer survivorship. This article discusses the role of constitutional and somatic genetics in determining outcome and survivorship following a diagnosis of cancer using illustrative examples primarily from the hematologic malignancies. PMID- 18395150 TI - Second primary cancers: an overview. AB - Substantial improvements in the past few decades in cancer detection and supportive care along with advances in therapy have led to growing numbers of cancer survivors. In view of the prolongation of survival in increasing numbers of patients, identification and quantification of the late effects of cancer and its therapy have become critical. One of the most serious events experienced by cancer survivors is the diagnosis of a new cancer. The number of patients who have second or higher-order cancers is increasing, and solid tumors are a leading cause of mortality among several populations of long-term survivors, including patients who have Hodgkin lymphoma. The focus of this article is treatment associated malignancies in survivors of selected adult cancers. PMID- 18395151 TI - Fertility and sexuality in young cancer survivors who have adult-onset malignancies. AB - Post-treatment fertility and sexuality are important issues to be considered before treatment starts in young adult patients who have cancer. Most male cancer survivors who have malignancies typical for young adulthood preserve the potential to father a child. Fertility remains decreased in comparable females treated for cancer in reproductive years. Postdiagnosis sexual life is affected, more so in females than in males, and was found to be strongly associated with emotional aspects of partnership. Communication about fertility and sexuality must become an integrated part of cancer survivors' follow-up. PMID- 18395152 TI - Long-term cardiac and pulmonary complications of cancer therapy. AB - Cardiac complications resulting from chemotherapy and radiation pose a significant risk for morbidity and mortality to the cancer survivor. Cardiac side effects may progress over time and are a concern for patients treated during childhood. Long-term pulmonary complications are relatively infrequent, and acute respiratory effects of drugs (mostly bleomycin) or radiation normally resolve early after therapy. Although most cardiovascular risk statistics and clinical experience are derived from patients treated before 1985, the modern radiation approach that limits the exposure of the heart and reduces the total dose seems to attenuate the previously observed cardiovascular risk. Potential preventive measures for high-risk patients are of increasing interest but remain experimental. PMID- 18395154 TI - Integrative oncology: complementary therapies for cancer survivors. AB - Cancer survivors experience a wide range of symptoms during and following completion of treatment, and some of these symptoms may persist for years or even decades. While pharmacologic treatments relieve many symptoms, they too may produce difficult side effects. Complementary therapies are noninvasive, inexpensive, and useful in controlling symptoms and improving quality of life, and they may be accessed by patients themselves. Rigorous scientific research has produced evidence that acupuncture, massage therapy, music, and mind-body therapies effectively and safely reduce physical and emotional symptoms. These therapies provide a favorable risk-benefit ratio and permit cancer survivors to help manage their own care. PMID- 18395155 TI - Cancer survivorship: advocacy organizations and support systems. AB - When oncology evolved into a specialized field of medicine more than four decades ago, the primary goals of most cancer treatment included the extension of patients' life expectancies and the occasional hope for cure. Physicians were seen as the principal and solitary advocate for patients, and information regarding cancer diagnosis, treatment, and side effects was delivered or screened by a doctor. Patient education materials were scarce, formalized support systems were nonexistent, and the future was often difficult to define. Patient advocacy has since expanded to models of self, organizational, and public policy advocacy. This article provides examples of advocacy organizations and support systems that offer guidance to providers and patients throughout the continuum of cancer care and into longer-term survival. PMID- 18395156 TI - Cancer survivorship: facing forward. AB - In the last three decades, the number of cancer survivors in the United States has tripled and is growing by 2% each year. In 2004, there were an estimated 10.7 million cancer survivors (representing 3.5% of the United States population) with a concomitant effect on public health. The growing and heterogeneous population of cancer survivors provides important opportunities for clinical and epidemiologic research into cancer biology, long-term treatment effects, prevention, and interventional research. In this article, the authors briefly review the history of the efforts that served to coalesce efforts to champion survivorship research, identify future challenges, and provide a perspective on future recommendations. PMID- 18395153 TI - Promoting a healthy lifestyle among cancer survivors. AB - With improving longevity, the late-occurring adverse effects of cancer and its treatment are becoming increasingly apparent. As in other clinical populations, healthy lifestyle behaviors encompassing weight management, a healthy diet, regular exercise, and smoking cessation have the potential to reduce morbidity and mortality significantly in cancer survivors. This article addresses the strength of evidence for recommendations in areas of weight management, diet, exercise, and smoking cessation; and the current evidence examining the efficacy of various intervention approaches to promote health behavior changes among adult cancer survivors. PMID- 18395157 TI - What ever happened to the doctor's soul? PMID- 18395158 TI - Prognostication--the lost skill of medicine. AB - Making a prognosis is one of the primary functions of the medical profession. At the end of the nineteenth century prognostication took up approximately ten percent of medical textbooks, by 1970 this had fallen to nearly zero. Given medical technology's awesome ability to prolong the process and suffering of dying today's patients need to know their prognosis in order to make choices about their treatment options. Whilst precise predictions of the future are obviously not possible, relatively simple mathematical modelling techniques can make reasonable estimates of likely outcomes for individual patients. The life expectancy of a patient of any age with any illness can be estimated provided the disease-specific mortality of the illness is known. Decision analysis or logistic regression models can then be used to determine the risks and benefits of various treatment options. A patient's prognosis does not just depend on their age and primary diagnosis, but also on the severity of their illness, their functional capacity both prior to and during the illness and the number of co-morbidities also suffered from. Several predictive instruments have been developed to help simplify the prediction of the outcome of individual patients. There are conflicting reports on how these models compare with doctors' intuition--whatever their strengths and weaknesses it is unlikely that they worsen clinical judgement. Therefore, all doctors should become familiar with them and use them appropriately. PMID- 18395159 TI - Cardiac resynchronisation therapy: evidence based benefits and patient selection. AB - Despite the improvement in pharmacologic treatment of heart failure, many patients continue to have severe persistent symptoms, and their prognosis remains poor. One of the most recent advances in heart failure management is the concept of cardiac resynchronization therapy (CRT) with right and left ventricular pacing. Large clinical trials have demonstrated morbidity and mortality benefits of CRT in patients with moderate to severe drug refractory heart failure (New York Heart Association (NYHA) functional class III or IV), and ejection fraction < or = 35% with QRS duration > or = 120 ms. Despite the documented benefits, 20 30% of patients selected to have CRT do not respond to this treatment. Echocardiography will probably play a more important role in better selecting patients with mechanical dyssynchrony who are more likely to respond to CRT. This article reviews the available evidence for CRT as well as the way to select responders to this rather invasive therapy. PMID- 18395160 TI - Barriers to obesity treatment. AB - Obesity, one of the most prevalent health problems in the Western world, is a chronic and progressive condition. Therefore, as with other chronic diseases, patients with obesity require lifelong treatment. Long-term efficacy and effectiveness of obesity treatments is notoriously poor. This may in part be attributable to the substantial barriers that undermine long-term obesity management strategies. These can include lack of recognition of obesity as a chronic condition, low socioeconomic status, time constraints, intimate saboteurs, and a wide range of comorbidities including mental health, sleep, chronic pain, musculoskeletal, cardiovascular, respiratory, digestive and endocrine disorders. Furthermore, medications used to treat some of these disorders may further undermine weight-loss efforts. Lack of specific obesity training of health professionals, attitudes and beliefs as well as coverage and availability of obesity treatments can likewise pose important barriers. Health professionals need to take care to identify, acknowledge and address these barriers where possible to increase patient success as well as compliance and adherence with treatments. Failure to do so may further undermine the sense of failure, low self esteem and self efficacy already common among obese individuals. Addressing treatment barriers can save resources and increase the prospect of long-term success. PMID- 18395161 TI - Microscopic colitis: a common and an easily overlooked cause of chronic diarrhoea. AB - Microscopic colitis, comprising collagenous colitis and lymphocytic colitis, is characterised clinically by chronic watery diarrhoea, a macroscopically normal colonic mucosa where diagnostic histopathological features are seen on microscopic examination. The annual incidence of each disorder is 4-6/100,000 inhabitants, with a peak incidence in 60-70 year old individuals and a noticeable female predominance in collagenous colitis. The aetiology is unknown. Abdominal pain, weight loss, fatigue, and faecal incontinence are common symptoms in addition to chronic diarrhoea that impair the health-related quality of life of the patient. There is an association to other autoimmune disorders such as celiac disease, diabetes mellitus, thyroid disorders and arthritis. Budesonide is the best-documented short-term treatment, but the optimal long-term strategy needs further study. The long-term prognosis is good and the risk of complications including colonic cancer is low. PMID- 18395162 TI - Electrocardiographic QT interval and cardiovascular reactivity in fibromyalgia differ from chronic fatigue syndrome. AB - BACKGROUND: Fibromyalgia (FM) and chronic fatigue syndrome (CFS) frequently overlap clinically and have been considered variants of one common disorder. We have recently shown that CFS is associated with a short corrected electrocardiographic QT interval (QTc). In the present study, we evaluated whether FM and CFS can be distinguished by QTc. METHODS: The study groups were comprised of women with FM (n=30) and with CFS (n=28). The patients were evaluated with a 10 min supine-30 min head-up tilt test. The electrocardiographic QT interval was corrected for heart rate (HR) according to Fridericia's equation (QTc). In addition, cardiovascular reactivity was assessed based on blood pressure and HR changes and was expressed as the 'hemodynamic instability score' (HIS). RESULTS: The average supine QTc in FM was 417 ms (SD 25) versus 372 ms (SD 22) in CFS (p<0.0001); the supine QTc cut-off <385.7 ms was 79% sensitive and 87% specific for CFS vs. FM. The average QTc at the 10th minute of tilt was 409 ms (SD 18) in FM versus 367 ms (SD 21) in CFS (p<0.0001); the tilt QTc cut-off <383.3 ms was 71% sensitive and 91% specific for CFS vs. FM. The average HIS in FM patients was -3.52 (SD 1.96) versus +3.21 (SD 2.43) in CFS (p<0.0001). CONCLUSION: A relatively short QTc and positive HIS characterize CFS patients and distinguish them from FM patients. These data may support the contention that FM and CFS are separate disorders. PMID- 18395164 TI - Admission characteristics predicting longer length of stay among elderly patients hospitalized for decompensated heart failure. AB - BACKGROUND: Acutely decompensated heart failure (HF) has become the leading cause of hospitalization for people aged 65 or older. Hospital length of stay (LOS) is a key determinant of higher hospitals costs. The aim of our study is to identify the admission characteristics that predict a longer LOS for elderly patients admitted for an acute exacerbation of HF. METHODS: We prospectively evaluated 324 patients (65 years of age or older), who were consecutively admitted for decompensated HF to a tertiary teaching hospital. Variables present at the time of emergency room evaluation that could predict a longer hospital LOS were determined by comparing the characteristics of patients hospitalized for less than 4 days with those of patients needing a longer stay. RESULTS: There were 191 women (59%) and 133 men in the study, with an average age of 78.6 years and a mean LOS of 7.1 days. Multivariate regression models identified two independent predictors of a hospital stay longer than four days: female gender (p=0.03, OR 1.645, 95% CI 1.047-2.584) and poorer NYHA functional class (p<0.01, OR 1.699, 95% CI 1.135-2.542). CONCLUSION: In elderly patients admitted for decompensated HF, the female gender and a worse functional class at the time of admission were associated with a longer subsequent LOS. PMID- 18395163 TI - Clinical outcome in patients with peripheral artery disease. Results from a prospective registry (FRENA). AB - BACKGROUND: The risk of future cardiovascular events in patients with peripheral artery disease (PAD) is often underestimated. PATIENTS AND METHODS: FRENA is an ongoing, observational registry of consecutive outpatients with symptomatic PAD, coronary artery disease (CAD) or cerebrovascular disease (CVD). We compared the incidence of major cardiovascular events (i.e., myocardial infarction, ischemic stroke, critical limb ischemia, or cardiovascular death) during a 12-month follow up period in a series of consecutive outpatients with PAD, CAD or CVD. RESULTS: As of December 2006, 1265 patients had been enrolled in FRENA who completed the 12-month follow-up. Of these, 417 patients (33%) had PAD, 474 (37%) had CAD, 374 (30%) had CVD. Patients with PAD had an increased incidence of major cardiovascular events per 100 patient-years: 17 (95% CI: 13-22) vs. 7.9 (5.5-11) in those with CAD, or 8.9 (6.1-13) in those with CVD. Compared to patients with CAD or CVD those with PAD had a similar incidence of myocardial infarction or stroke, but a higher incidence of critical limb ischemia, limb amputation and death. This incidence increased with the severity of the symptoms: 8.7 (95% CI: 5.3-13) in patients in Fontaine stage IIa; 25 (95% CI: 16-38) in stage IIb; 26 (95% CI: 13-47) in stage III; 42 (95% CI: 24-67) in stage IV. CONCLUSIONS: Our data confirm a higher incidence of major cardiovascular events for patients with PAD, as well as a correlation of these events with the severity of PAD. PMID- 18395165 TI - Atorvastatin monotherapy vs. combination therapy in the management of patients with combined hyperlipidemia. AB - BACKGROUND: Mixed hyperlipidemia is a common disorder characterized by elevated VLDL and LDL levels. Patients with this syndrome usually are in need of combination therapy, comprising a fibric acid derivate with a statin drug in order to achieve LDL and triglyceride target values. Atorvastatin is a hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitor demonstrated to be effective in reducing both cholesterol (CHOL) and triglyceride (TG) levels in humans. We examined the efficacy of atorvastatin as monotherapy in achieving a better or the same lipid profile in patients with mixed hyperlipidemia treated with combination therapy. DESIGN: We compared atorvastatin with a combination of a fibric acid derivate and a statin drug (other than atorvastatin) in a 24-week, prospective randomized, open-label study of 27 patients with mixed hyperlipidemia. METHODS: All 27 patients had been treated with statin-fibrate therapy in different regimens for at least a year. Atorvastatin at a daily dose of 20 mg was substituted for statin-fibrate therapy. Lipid and safety profiles were assessed. RESULTS: Atorvastatin significantly reduced total cholesterol, LDL C, and HDL-C compared to statin-fibrate therapy. In contrast, TG and glucose levels were significantly elevated with atorvastatin. Target LDL-C and TG was achieved in 10 patients with the single therapy of atorvastatin vs. 6 patients under statin-fibrate. In 16 patients, atorvastatin was at least as effective as, or better than, the combination therapy, and was recommended for continuation of treatment. CONCLUSION: Atorvastatin is an adequate monotherapy for many mixed hyperlipidemia patients. We recommend atorvastatin be considered for every patient suffering from mixed hyperlipidemia. PMID- 18395166 TI - Classical cardiovascular risk factors according to fasting plasma glucose levels. AB - BACKGROUND: To compare the prevalence of classical cardiovascular risk factors (CVRF) and metabolic syndrome (MetS) in our population according to fasting plasma glucose levels (FPG). METHODS: We have studied 344 subjects between 20-70 years of age, recruited in a Primary Care Clinic. Subjects were divided into four groups according to their fasting plasma glucose (FPG) values: normal plasma glucose (NG) when FPG < 5.6 mmol/L; FPG between 5.6 and 6.0 mmol/L (FPG1); FPG between 6.1-6.9 mmol/L (FPG2); and diabetes (DM) FPG > or = 7 mmol/L or previous diagnosis of diabetes. Cardiovascular risk factors (hypertension, TC/HDL-C index and Apo B values), presence of the MetS and indirect measure of insulin resistance (HOMA) were analyzed. RESULTS: Subjects with FPG2 have a prevalence of classic CVRF and MetS similar to that observed in subjects with type 2 diabetes mellitus (T2DM). The TC:HDL-C index > or = 5 in 56% and 57%, Apo B > or = 1.2 g/L in 59% and 57%, hypertension in 60% and 54% of FPG2 and T2DM subjects, respectively. MetS was diagnosed in 79% of FPG2 and 80% of T2DM. We found significant differences with FPG1 group who presented low CVRF and MetS proportion. CONCLUSION: In our population FPG2 and T2DM subjects show a similar cardiovascular risk profile. On the other hand, such risk is significantly lower in subjects with FPG between 5.6-6.0 mmol/L. These results might have practical implications. PMID- 18395167 TI - Pulmonary embolism associated with the use of anabolic steroids. AB - We present the case of a 56-year-old man with deep vein thrombosis (DVT) and pulmonary embolism (PE). He had been given intramuscular injections of testosterone and the anabolic-androgenic steroid nandrolone, due to a muscle injury, a total of three times prior to manifestation of the symptoms. An ultrasonographic examination of the right leg revealed a DVT and computed tomography of the pulmonary arteries showed PE. The thromboembolic episodes in this previously healthy patient were in all probability associated with intramuscular injections of testosterone and nandrolone, to which there is a clear correlation in time. PMID- 18395168 TI - Diagnostic pitfall: Mycobacterium avium complex pulmonary infection and positive ANCA. AB - We report the case of a 58-year-old female who presented with productive cough, weight loss, pulmonary nodular infiltrates and cavitations. She had a positive anti-neutrophil cytoplasmic antibodies (ANCA) test. A diagnosis of vasculitis was considered and a video-assisted thoracoscopic biopsy of the lung nodules was organised. However, prior to the biopsy, the sputum results revealed the presence of acid-fast bacilli, which were identified as Mycobacterium avium complex. A repeat ANCA assay was positive for atypical ANCA with negative proteinase-3 and myeloperoxidase titres. The patient was treated with rifampicin, ethambutol and clarithromycin with clinical and radiological improvement. The objective of this report is to highlight a rare association between positive ANCA titres and a non tuberculous mycobacterial infection as a misdiagnosis and treatment of this patient with immunosuppressive therapy might have led to serious consequences. PMID- 18395169 TI - Stridor secondary to hypocalcemia in the elderly: an unusual presentation. AB - Though hypocalcemic stridor is a well-recognized pediatric emergency, it has rarely been reported in the elderly, and hypocalcemia of nutritional origin presenting with stridor in this group has not previously been reported. We report an elderly patient who presented with stridor and intermittent sudden airway obstruction resulting from laryngospasm secondary to hypocalcemia of nutritional origin and we present a brief review of this life-threatening complication of hypocalcemia. PMID- 18395170 TI - Interference in free thyroxine immunoassay. AB - We report two patients who presented with non-specific symptoms of tiredness that prompted their clinician to order a thyroid function test. In both cases, free thyroxine (T4) was reported to be high, with a normal thyroid-stimulating hormone (TSH). Repeated blood tests from the same laboratory using the same assay method yielded similar results. Due to discrepancies between the clinical and laboratory findings, we decided to repeat the blood tests at another laboratory using a different assay method. This revealed normal free T4 and TSH levels, suggesting that heterophile antibodies were interfering with the free T4 assay, leading to clinical confusion. PMID- 18395171 TI - Susac's syndrome presenting with consciousness disturbance. PMID- 18395173 TI - Comments on "Isolated hypoaldosteronism: an overlooked cause of hyponatraemia". PMID- 18395174 TI - Polypharmacy: an undervalued component of complexity in the care of elderly patients. PMID- 18395175 TI - How accurate are diagnostic coding data? PMID- 18395176 TI - Kaposi's sarcoma in an immunocompetent patient following corticosteroid therapy. PMID- 18395177 TI - Malaria research, 1980-2004, and the burden of disease. AB - Malaria is estimated to cause about 1.6% of the 57 million deaths occurring annually and 2.3% of the disease burden. However, it accounts for only about 0.4% of world biomedical research, and this percentage is barely changing. Most of the research takes place in Europe and North America, which are little affected directly by the disease, 90% of whose burden occurs in sub-Saharan Africa. Research includes both pharmaceutical and non-pharmaceutical approaches; the fastest growing ones involve the artemisinins and genetics. Leading countries in malaria research (including India, Thailand, Kenya and Nigeria) differ greatly in the subjects that they favour. PMID- 18395178 TI - Seroepidemiology and serological follow-up of anti-leptospiral IgG in children in Southern Vietnam. AB - A follow-up study was conducted with 23 months interval to investigate the seroepidemiology and persistence of Leptospira IgG antibodies among healthy children in Binh Thuan province, Southern Vietnam. Sera from 262 children (7-13 years of age) were collected and analysed with a commercially available enzyme linked immunosorbent assay (ELISA) for Leptospira IgG. Seroconversion was observed in 10.4% (22 of 211, 95% CI: 5.6-26.7) of the children, of whom 18 (8.5%) had probably and four (1.9%) had certainly been exposed to Leptospira. Based on the reduction of sero-negatives of 1.9% among children who have been certainly exposed, the annual seroconversion rate, a measure of the incidence rate of Leptospira infections, corresponds to 0.99% (95% CI: 0.39-2.52). In 61% (31 of 51, 95% CI: 47.1-73.0) of the children with past-infection, Leptospira IgG antibodies remain detectable after 2 years. Data from this study indicate that IgG antibody responses against Leptospira may persist at least for 2 years in children without manifestations of leptospirosis. Results of study uncover the true incidence of leptospirosis infection, the dynamics of waxing and waning antibody concentrations and points at a larger burden of clinically non significant Leptospira infections in Southern Vietnam. This also indicates background reactivity for serological testing and thus serological result of a single serum sample must be carefully interpreted. PMID- 18395179 TI - [Staphylococcus aureus bacteremia and endocarditis]. AB - The prevalence of Stapylococcus bacteriaemia is increasing worldwide, because of the increasing use of invasive procedures leading to nosocomial infections, but also of a changing way of life (increasing fashion for tattoos or piercing, use of intravenous drugs). Infective endocarditis develops in 10-30% of the cases of staphylococcus bacteriaemia. Staphylococcus aureus endocarditis must be suspected when it develops in the year following heart surgery or implantation of permanent devices. In drug users, it usually involves the tricuspid valve. According to the resistance of the germ to meticillin, antibiotic therapy uses a combination of intravenous penicillin or glycopeptide and an aminoside. Other antibiotics such as fosfomycin, rifampicin, fusidic acid, or clindamycin can be used when aminosides are contra-indicated. The role of newer antibiotic agents, such as daptomycin or linezolide, remains to be established. PMID- 18395180 TI - [New anti-Gram+antibiotics: which role in infective endocarditis?]. AB - Though the increased resistance to antibiotics observed worldwide is not a major concern in France, treatment of methicillin-resistant S. aureus has important limitations. New antibiotics have recently been marketed or will soon be (quinupristin-dalfopristin, linelozide, tigecyclin, daptomycin, ceftobiprole, dalbavancin). Their role, which has been documented in several forms of acute infections, remains to be established in infective endocarditis. At present, only treatment of methicillin-resistant S. aureus right-sided endocarditis justifies the use of daptomycin, preferably in association with rifampicin, when the use of vancomycin is not possible or contraindicated. PMID- 18395181 TI - [New microbiologic diagnostic techniques in infective endocarditis]. AB - The new techniques used for identifying causal germs in infective endocarditis are reviewed. Techniques such as polymerase chain reaction are now used to identify previously unrecognised germs. Choosing the appropriate microbiologic technique requires close collaboration between clinicians and microbiologists. PMID- 18395182 TI - Ectopic hyperprolactinaemia in a woman with a mesocolic perivascular epithelioid cell tumor ("PEComa"). AB - Ectopic prolactin secretion remains exceptional and originates mainly from malignant tumors. We report the case of a 47-year-old woman who presented amenorrhea leading to unravel important hyperprolactinaemia (269 ng/mL) with no hypothalamo-pituitary mass on magnetic resonance imaging (MRI). Pelvic imaging revealed the presence of a large pelvic mass that originated from the mesocolon. After complete surgical extraction, histological examination was in favour of a "perivascular epithelioid cell tumor" (PEComa). Prolactin levels normalized after surgical extraction and remained normal after a 3-year follow-up, totally free of tumour recurrence and/or metastasis. This suggests that hyperprolactinaemia was most likely related to the PEComa, despite negative reactions with antiprolactin antibodies at immunohistochemistry. Alternatively to a direct prolactin secretion by the tumor, one could hypothesize that the tumour secreted a prolactin stimulating factor or a dopamine antagonist that could not be identified. In conclusion, in face of an important hyperprolactinaemia without any hypothalamic pituitary mass, it remains important to search for an ectopic prolactin production, such as a PEComa. PMID- 18395183 TI - Sleep and errors in a group of Australian hospital nurses at work and during the commute. AB - There is a paucity of information regarding Australian nurses' sleep and fatigue levels, and whether they result in impairment. Forty-one Australian hospital nurses completed daily logbooks for one month recording work hours, sleep, sleepiness, stress, errors, near errors and observed errors (made by others). Nurses reported exhaustion, stress and struggling to remain (STR) awake at work during one in three shifts. Sleep was significantly reduced on workdays in general, and workdays when an error was reported relative to days off. The primary predictor of error was STR, followed by stress. The primary predictor of extreme drowsiness during the commute was also STR awake, followed by exhaustion, and consecutive shifts. In turn, STR awake was predicted by exhaustion, prior sleep and shift length. Findings highlight the need for further attention to these issues to optimise the safety of nurses and patients in our hospitals, and the community at large on our roads. PMID- 18395184 TI - Comprehensive change management concepts. Development of a participatory approach. AB - During the last years, many change projects in organizations did not have the planned success. Therefore at first, the causes for these failures and the success factors contributing to organizational change have to be discussed. To get better results, a comprehensive change management concept has been developed and tested in an ongoing research project. By using concepts for an integrated assessment and design of organizations, an approach for analyzing the current situation has been elaborated to identify "lack of integration" in the change initiatives of a company. To realize an integrated overall approach of modernization by harmonizing different methods and concepts, first, one has to prove their relationship to policy and strategy (vertical harmonization). The second step is to take into account the fact that there has to be a logical fit between the single concepts (horizontal harmonization). But even if all elements are logically coherent, that does not mean that the people working in the company also see this coherence. Therefore, in addition to the "logical fit", one has to examine the "psychological fit". In the end, a concept for analyzing the status quo in an organization as a result of "objective data" and "subjective data" originated. Subsequently, instruments for harmonizing different modernizing concepts have been applied. As part of the comprehensive change management concept participatory ergonomic approaches have been used during the project. The present study shows this approach in the case of one company. PMID- 18395185 TI - Effects of threat cues on attentional shifting, disengagement and response slowing in anxious individuals. AB - According to cognitive models of anxiety, attentional biases for threat may cause or maintain anxiety states. Previous research using spatial cueing tasks has been interpreted in terms of difficulty in disengaging attention from threat in anxious individuals, as indicated by contrasts of response times (RTs) from threat cue versus neutral cue trials. However, on spatial cueing tasks, differences in RT between threat cue and neutral cue trials may stem from a slowing effect of threat on RT, as well as effects on allocation of visuospatial attention. The present study examined the effects of threat cues on both attentional cueing and response slowing. High and low anxious individuals completed a central cue task, which assessed threat-related response slowing, and a spatial cueing task, which assessed attentional biases for angry, happy and neutral faces. Results indicated that interpretation of the anxiety-related bias for threat depended on whether the effect of response slowing was taken into account. The study illustrates an important problem in using the modified spatial cueing task to assess components of threat-related attentional bias. As this experimental method may reflect both threat-related attentional cueing and response slowing effects, it cannot be assumed to provide pure measures of shift or disengagement components of attention bias. PMID- 18395186 TI - Alterations in S-adenosylhomocysteine metabolism decrease O6-methylguanine DNA methyltransferase gene expression without affecting promoter methylation. AB - The DNA repair enzyme O(6)-methylguanine DNA methyltransferase (MGMT) protects cells against the cytotoxic effects of alkylating agents. Therefore, modulation of MGMT expression in tumors is a possible strategy for improving the efficiency of cancer therapy. MGMT expression and activity is lost frequently in association with DNA hypermethylation of the MGMT promoter region. Since DNA and mRNA methylation are controlled by intracellular S-adenosylmethionine (AdoMet) and S adenosylhomocysteine (AdoHcy) levels, we hypothesized a role for AdoMet/AdoHcy ratio in the regulation of MGMT promoter methylation and mRNA expression. Our initial studies showed that AdoMet/AdoHcy ratios vary over a wide range (7.0-50) in different glioblastoma and hepatoma cell lines. The studied cell lines exhibit distinct MGMT promoter methylation patterns: MGMT promoter was completely unmethylated in LN-18 and Tu 132 cells, hypermethylated in LN-229, U87-MG, and Tu 113 cells, and partially methylated in HepG2 cells. Furthermore, MGMT promoter methylation patterns and global DNA methylation are not related to intracellular AdoMet/AdoHcy ratio under control conditions. To lower AdoMet/AdoHcy ratio to values <1 we used AdoHcy hydrolase inhibitor adenosine-2',3'-dialdehyde (30 microM) and found that neither short-term (24 h) nor long-term changes (7 weeks) in AdoMet/AdoHcy ratio altered global or MGMT promoter methylation. However, experimentally elevated AdoHcy levels significantly decreased MGMT mRNA levels by >50% in all MGMT-expressing cell lines, which is most likely the result of impaired mRNA methylation. Thus, the present study suggests elevation of AdoHcy levels by AdoHcy hydrolase inhibition as a novel pharmacological approach to modulate MGMT expression and to increase the responsiveness to alkylating agents. PMID- 18395187 TI - Inhibition of caspase-3 activity and activation by protein glutathionylation. AB - Protein glutathionylation is a post-translational modification that may account for a broad mechanism of redox signaling. The caspase family of cysteine proteases represents a potential target for regulation by glutathionylation. To examine this, caspase proteins, derived from HL-60 cells after activation with actinomycin D, were incubated with GSSG. Total protein glutathionylation was enhanced and caspase-3 activity was inhibited in a dose- and time-dependent manner by GSSG. Caspase inhibition was reversible by thiol-specific reducing reagents. Proteolytic activation of caspases was also affected, as the activation of procaspase-3 and procaspase-9 in HL-60 cell extracts induced by cytochrome c and dATP was inhibited by pre-incubation with GSSG. When biotin-labeled GSSG was incubated with recombinant caspase-3, biotin label was found associated with both p12 and p17 subunits of active caspase-3 by non-reducing SDS-PAGE. Caspase-3 glutathionylation was confirmed by matrix assisted laser desorption ionization (MALDI) mass spectrometric analysis of GSSG-treated recombinant caspase-3. Specific sites of glutathionylation were identified as Cys(135) of the p17 protein (the active site) and Cys(45) of the p12 protein. These results indicate that glutathionylation of caspase can occur at physiologically relevant concentrations of GSSG and results in the inhibition of caspase activation and activity. PMID- 18395188 TI - Up-regulation of pro-nerve growth factor, neurotrophin receptor p75, and sortilin is associated with retrovirus-induced spongiform encephalomyelopathy. AB - The progressive spongiform encephalomyelopathy caused by ts1, a neuropathogenic temperature-sensitive mutant of Moloney murine leukemia virus (MoMuLV-ts1), results in motor neuronal loss without direct neuronal infection. We have previously reported that ts1-mediated neuronal degeneration in mice has a multifactorial pathogenesis. Here, we report that in the ts1-infected central nervous system (CNS) activated neural cells showed intense immunoreactivity for pro-nerve growth factor (proNGF), neurotrophin receptor p75 (p75(NTR)), and sortilin in the areas showing spongiform changes. Since recent studies suggested that proNGF is more active than mature NGF in inducing neuronal death after binding to co-receptors p75(NTR)/sortilin, we hypothesized that overexpression of proNGF, sortilin and p75(NTR) play a role in ts1-induced neurodegeneration. We found that proNGF and p75(NTR), but not sortilin, mRNA and protein were significantly elevated in ts1-infected brainstem compared to non-infected control tissue. There was extensive tyrosine phosphorylation of p75(NTR), a marker for its activation, in ts1-infected brainstem with abundance in degenerating neurons. We explored whether the increase in the in vivo proNGF expression also occurs in cultured immortalized C1 astrocytes infected by ts1 virus. The proNGF level was significantly increased in infected C1 cells compared to control cells only after addition of fibroblast growth factor (FGF-1). We also showed increased expression of FGF-1 in the CNS of ts1-infected mice. Our findings suggest that the FGF-1 signaling pathway may be responsible for the overexpression of proNGF in neural cells during pathogenesis of ts1-induced neurodegeneration. This study provides new in vivo insights into the possible role of proNGF and its receptors in ts1 induced neurodegeneration. PMID- 18395190 TI - Unmyelinated primary afferents from adjacent spinal nerves intermingle in the spinal dorsal horn: a possible mechanism contributing to neuropathic pain. AB - Peripheral nerve injury in animals can cause neuropathic pain often expressed in the form of hyperalgesia and allodynia. Spinal nerve ligation, in which the fifth and sixth lumbar (L5 and L6) or only the L5 spinal nerve is ligated and cut, is a model commonly used to produce neuropathic pain. The purpose of the present study was to test whether there is any anatomical evidence to support the suggestion that terminating unmyelinated (C) fibres of injured and adjacent uninjured nerves interact at the level of the spinal dorsal horn. Thus, in the first series of experiments, rats received injections of anterograde tracers, either wheat germ agglutinin (WGA) conjugated to horseradish peroxidase or Bandeiraea simplicifolia isolectin B4 (IB4), into the L4 or L5 spinal nerves. Results with both tracers showed that the central terminals of nerve L4 were concentrated in both L4 and L3 segments of the dorsal horn with clear although reduced levels of labelling in L2 and L5. Similarly, the central terminals of nerve L5 were found in both L5 and L4 again with less labelling in L3 and L6. These results suggest an intermingling of primary afferents of adjacent nerves at the level of the spinal dorsal horn. A second series of experiments was therefore conducted to test whether primary afferent terminals from adjacent nerves target the same neuronal elements in the regions of overlap. Consequently, additional rats were injected with WGA into the L5 spinal nerve and IB4 into the adjacent L4 spinal nerve. Double immunofluorescent staining and confocal microscopy revealed that IB4-labelled and WGA-labelled boutons, belonging to L4 and L5 spinal nerves, terminated in the same region within the L4 spinal segment. This suggests that neurons located in regions of overlap receive input from both L4 (intact) and L5 (injured) afferents. Consequently, spinal neurons located in regions of terminal overlap may show augmented responses to activation of the intact L4 nerve due to neuronal sensitisation resulting from injury to the adjacent L5 nerve. This may in part provide an anatomical basis for hyperalgesic reaction to injury. PMID- 18395189 TI - Purinergic P2X receptors presynaptically increase glutamatergic synaptic transmission in dorsolateral periaqueductal gray. AB - Purinergic P2X receptors have been reported to be present in regions of the midbrain periaqueductal gray (PAG). The purpose of this study was to determine the role of presynaptic P2X receptors in modulating excitatory and inhibitory synaptic inputs to the dorsolateral PAG (dl-PAG), which has abundant neuronal connections. First, whole cell voltage-clamp recording was performed to obtain excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) of the dl-PAG neurons. Our data show that alpha, beta-methylene ATP (a P2X receptor agonist), in the concentration of 50 microM, significantly increased the frequency of miniature EPSCs without altering the amplitude of miniature EPSCs in eight tested neurons. The effects were attenuated by PPADS, an antagonist to P2X receptors. Furthermore, alpha, beta-methylene ATP increased the amplitude of evoked EPSCs, and decreased the paired-pulse ratio of eEPSCs in ten neurons. In contrast, activation of P2X had no distinct effect on IPSCs. In addition, immunofluorescent methods demonstrate that P2X labeling was co-localized with a presynaptic marker, synaptophysin, in the dl-PAG. The results of the current study provide the first evidence indicating that P2X receptors facilitate glutamatergic synaptic transmission in the dl-PAG via presynaptic mechanisms. PMID- 18395192 TI - Central projections of the sensory innervation of the rat middle meningeal artery. AB - Headaches, especially migraine, involve not only pain but also aspects such as vasodilation of cranial vessels and sensitization of nerve endings, processes dependent on and connected to the central nervous system. To understand pathogenic mechanisms of headache, it is important to elucidate the central projections of sensory nerves that innervate cranial vessels, of which the middle meningeal artery (MMA) is the largest artery supplying the dura mater. In this study, cholera toxin subunit b (CTb) or wheat germ agglutinin-horseradish peroxidase conjugate (WGA-HRP) was applied on the adventitia of MMA. After perfusion fixation, the brainstem, the C1-C4 spinal segments and the trigeminal and C2 dorsal root ganglia were removed and sections from these tissues were processed to visualize transported tracers. Labeled cell bodies were seen ipsilaterally in the trigeminal and C2 dorsal root ganglia. Labeled nerve terminations were found ipsilaterally in the lateral part of the spinal dorsal horn of segments C1-C3 and in the caudal and interpolar parts of the spinal trigeminal nucleus. WGA-HRP labeled terminations were mainly located in laminae I and II, whereas CTb labeled terminations located in laminae III-V. These results indicate that sensory information from the MMA is transmitted through both trigeminal and cervical spinal nerve branches to a region in the central nervous system extending rostrally from the C3 dorsal horn to the interpolar part of the spinal trigeminal nucleus. Our data further substantiates that the sensory innervation of the MMA, in addition to putative nociceptive afferents, include a population of large caliber afferents with an as yet unclear but presumably non nociceptive role. PMID- 18395191 TI - Reelin, a guidance signal for the regeneration of the entorhino-hippocampal path. AB - The importance of reelin for cortical lamination in the developing CNS is well established, but its role in nerve fiber growth is not well understood. In this study, hippocampal slices were co-cultured with entorhinal slices of GFP mice, in order to compare the growth of the entorhino-hippocampal path in wild type and reeler mice. On day 6, regenerated fibers were seen to navigate from the entorhinal cortex into the hippocampus. The results showed that in wild type mice, regenerated fibers grew along the molecular layer of the dentate gyrus, and only a few fibers were found to penetrate through the granular layer into the hilus. This specific orientation was similar to the perforant path in vivo. Compared with perforant path regeneration in wild type mice, reeler mice seemed to have lost their specific orientation and proper termination in the hippocampus. Without the guidance signal from reelin, the regenerated fibers left the molecular layers and continued to grow aberrantly, i.e., in the granular layer, hilus, pyramidal layer and even stratum oriens. Particularly in the dentate gyrus, the fibers meandered around the cells in the hilus and resembled a network. The study concludes that reelin also serves as an important guidance signal for neuroregeneration of the perforant path. PMID- 18395193 TI - Induction of BiP, an ER-resident protein, prevents the neuronal death induced by transient forebrain ischemia in gerbil. AB - Endoplasmic reticulum (ER) stress, which is caused by the accumulation of unfolded proteins in the ER lumen, is associated with stroke and neurodegenerative diseases such as Parkinson's and Alzheimer's diseases. We evaluated the effect of a selective inducer of immunoglobulin heavy chain binding protein (BiP) (BiP inducer X; BIX) against both tunicamycin-induced cell death (in SH-SY5Y cells) and the effects of global transient forebrain ischemia (in gerbils). BIX significantly induced BiP expression both in vitro and in vivo. Pretreatment with BIX at 2 or 5 microM reduced the cell death induced by tunicamycin in SH-SY5Y cells. In gerbils subjected to forebrain ischemia, prior treatment with BIX (intracerebroventricular injection at 10 or 40 microg) protected against cell death and decreased TUNEL-positive cells in the hippocampal CA1 subfield. These findings indicate that this selective inducer of BiP could be used to prevent the neuronal damage both in vitro and in vivo. PMID- 18395194 TI - Glia maturation factor modulates beta-amyloid-induced glial activation, inflammatory cytokine/chemokine production and neuronal damage. AB - Glia maturation factor (GMF), discovered and characterized in our laboratory, is a highly conserved protein primarily localized in mammalian central nervous system. Previously we demonstrated that GMF is required in the induced production of proinflammatory cytokines and chemokines in brain cells. We now report that ventricular infusion of human amyloid beta peptide1-42 (Abeta1-42) in mouse brain caused glial activation and large increases in the levels of GMF as well as induction of inflammatory cytokine/chemokine known for launching the neuro inflammatory cascade in Alzheimer's disease (AD). To test the hypothesis that GMF is involved in the pathogenesis of AD, we infused Abeta1-42 in the brain of GMF deficient (GMF-KO) mice, recently prepared in our laboratory. GMF-deficient mice showed reduced glial activation and significantly suppressed proinflammatory cytokine/chemokine production following Abeta infusion compared to wild type (Wt) mice. The decrease in glial activation in the GMF-KO mice is also associated with significant reduction in Abeta induced loss of pre-synaptic marker, synaptophysin, and post-synaptic density protein-95 (PSD 95). We also examined the potential relationship between GMF or lack of it with learning and memory using the T-maze, Y-maze, and water maze, hippocampal-dependent spatial memory tasks. Our results show that memory retention was improved in GMF-KO mice compared to Wt controls following Abeta infusion. Diminution of these Abeta1-42 effects in primary cultures of GMF-KO astrocyte and microglia were reversed by reconstituted expression of GMF. Taken together, our results indicate a novel mediatory role of GMF in the neuro-inflammatory pathway of Abeta and its pro inflammatory functions. PMID- 18395195 TI - Morphological and functional changes of blood-brain barrier in kindled rats with cortical dysplasia. AB - Cortical dysplasia (CD) is one of the major causes contributing to epileptogenesis associated with blood-brain-barrier (BBB) disturbances. The current study investigated the functional and ultrastructural changes of BBB in pentylenetetrazole (PTZ)-kindled rats with CD. Pregnant rats on E17 were exposed to 145 cGy of gamma-irradiation and offspring were used for experiments. The rats were given PTZ three times per week to induce kindling. The permeability of BBB was determined by using sodium fluorescein (NaFlu). Immunohistochemistry for occludin, GFAP and c-fos, western-blot analysis for occludin and electron microscopy for the ultrastructural alterations in BBB were performed. The brain level of NaFlu did not increase in rats with CD and/or kindling. Following administration of a convulsive dose of PTZ, a significant increase in BBB permeability was observed in kindled rats with CD. Occludin immunoreactivity and expression remained essentially unchanged in all groups. Slightly enhanced immunoreactivity for GFAP was observed in all groups except control. c-fos immunoreactivity in brain sections of kindled rats with CD displayed a striking increase by convulsive PTZ challenge. Tight junctions were ultrastructurally intact, whereas markedly increased number of pinocytotic vesicles was noted in brain endothelium of kindled rats with CD by convulsive dose of PTZ. The present study showed that epileptic seizures induced by convulsive PTZ challenge during kindling-mediated epileptogenesis in the presence of CD changed both functional and ultrastructural properties of the BBB and considerably enhanced transendothelial vesicular transport, while paracellular pathway was apparently not involved in this setting. PMID- 18395196 TI - Transitive and pseudo-transitive inferences. AB - Given that A is longer than B, and that B is longer than C, even 5-year-old children can infer that A is longer than C. Theories of reasoning based on formal rules of inference invoke simple axioms ("meaning postulates") to capture such transitive inferences. An alternative theory proposes instead that reasoners construct mental models of the situation described by the premises in order to draw such inferences. An unexpected consequence of the model theory is that if adult reasoners construct simple models of typical situations, then they should infer transitive relations where, in certain cases, none exists. We report four studies corroborating the occurrence of these "pseudo-transitive" fallacies. Experiment 1 established that individuals' diagrams of certain non-transitive relations yield transitive conclusions. Experiment 2 showed that these premises also give rise to fallacious transitive inferences. Experiment 3 established that when the context suggested alternatives to the simple models, the participants made fewer errors. Experiment 4 showed that tense is an important aspect of meaning which affects whether individuals draw transitive conclusions. We discuss the implications of these results for various theories of reasoning. PMID- 18395197 TI - A novel mRNA 3' untranslated region translational control sequence regulates Xenopus Wee1 mRNA translation. AB - Cell cycle progression during oocyte maturation requires the strict temporal regulation of maternal mRNA translation. The intrinsic basis of this temporal control has not been fully elucidated but appears to involve distinct mRNA 3' UTR regulatory elements. In this study, we identify a novel translational control sequence (TCS) that exerts repression of target mRNAs in immature oocytes of the frog, Xenopus laevis, and can direct early cytoplasmic polyadenylation and translational activation during oocyte maturation. The TCS is functionally distinct from the previously characterized Musashi/polyadenylation response element (PRE) and the cytoplasmic polyadenylation element (CPE). We report that TCS elements exert translational repression in both the Wee1 mRNA 3' UTR and the pericentriolar material-1 (Pcm-1) mRNA 3' UTR in immature oocytes. During oocyte maturation, TCS function directs the early translational activation of the Pcm-1 mRNA. By contrast, we demonstrate that CPE sequences flanking the TCS elements in the Wee1 3' UTR suppress the ability of the TCS to direct early translational activation. Our results indicate that a functional hierarchy exists between these distinct 3' UTR regulatory elements to control the timing of maternal mRNA translational activation during oocyte maturation. PMID- 18395198 TI - Disp1 regulates growth of mammalian long bones through the control of Ihh distribution. AB - Dispatched1 (Disp1) is required for the release of cholesterol modified hedgehog (Hh) proteins from producing cells. We investigated the role of Disp1 in Indian hedgehog (Ihh) signaling in the developing bone bypassing the lethality of the Disp1(C829F) allele at early somite stages through the supply of non-cholesterol modified Sonic hedgehog (N-Shh). The long bones that develop in the absence of wild-type Disp1, while clearly shorter, have a juxtaposition of proliferating and non-proliferating hypertrophic chondrocytes that is markedly more normal in organization than those of ihh null mutants. Direct analysis of Ihh trafficking in the target field demonstrates that Ihh is distributed well beyond Ihh expressing cells though the range of movement and signaling action is more restricted than in wild-type long bones. Consequently, a PTHrP-Ihh feedback loop is established, but over a shorter distance, reflecting the reduced range of Ihh movement. These analyses of the Disp1(C829F) mutation demonstrate that Disp1 is not absolutely required for the paracrine signaling role of Ihh in the skeleton. However, Disp1 is critical for the full extent of signaling within the chondrocyte target field and consequently the establishment of a normal skeletal growth plate. PMID- 18395199 TI - Fish consumption and PCB-associated health risks in recreational fishermen on the James River, Virginia. AB - Consumption of sport-caught fish contaminated with high levels of polychlorinated biphenyls (PCBs) may pose human health risks. To obtain estimates of fish consumption and fishing behaviors in recreational fishermen in Virginia, on-site interviews (n=143; 134 men and 9 women) were conducted at seven public boat landings along the James River. Using existing PCB concentration data from James River fish tissue samples collected from 1997, and 1999 to 2001, default and point estimates were calculated and Monte Carlo analyses conducted to estimate potential risks under different consumption scenarios. A mean of 55 fish meals/yr and 14 James River sport-caught fish (JRSCF) meals/yr were reported. Caucasians fished less often (mean of 58 d), consumed less fish (mean of 43 meals/yr) and had smaller portion sizes (mean of 11.7 oz) compared to other races combined (130 d; 82 meals/yr; and 15.6 oz). On average, respondents reported consuming 10 meals of James River catfish a year (5 kg/yr). Risk estimates produced from Monte Carlo analysis were consistently lower than the default and point estimates. Several individuals exceeded acceptable risk levels and the mean cancer and non-cancer risks among catfish consumers exceeded acceptable levels. Eighteen percent of individuals had no knowledge of fish advisories in Virginia and 4% of the subjects indicated they would consume fish under advisory. Based on reported consumption, a significant risk to recreational fishermen, as a result of consuming PCB-contaminated catfish, was found. Risks associated with consuming other species were within acceptable limits. PMID- 18395200 TI - Diabetes-, stress- and ageing-related changes in synaptic plasticity in hippocampus and neocortex--the same metaplastic process? AB - Learning and memory in the brain likely occur through activity-dependent, long lasting changes in synaptic transmission. Two opposite activity-dependent synaptic modifications have been identified so far, long-term potentiation and long-term depression. In many brain areas including hippocampal CA1 and neocortex, the level of postsynaptic depolarization controls the magnitude and sign of plasticity: long-term depression is obtained after low depolarizations, whereas long-term potentiation requires stronger ones. Synaptic plasticity also depends on prior synaptic activity. Activity-dependent modulation of subsequent induction of synaptic plasticity, termed "priming" or "metaplasticity", is due, at least in part, to concomitant opposite shifts in the levels of postsynaptic depolarization needed to elicit synaptic plasticity: in previously activated or potentiated synapses, induction of long-term potentiation requires a larger depolarization and that of long-term depression a smaller one compared with naive synapses - i.e. potentiation is inhibited and depression promoted - and vice versa in depressed synapses. Many species including humans express cognitive deficits during ageing, diseases (diabetes mellitus, ...) and psychological insults (stress, ...). Interestingly, diabetic, stressed and aged rats show robust long-term depression and long-term potentiation. But, as in metaplasticity, induction of long-term potentiation requires a larger postsynaptic depolarization and that of long-term depression a smaller one compared with young control animals. Moreover, diabetes- and activity-dependent modulation of synaptic plasticity exhibit occlusion. This suggests that diabetes, stress and ageing act on synaptic plasticity through common mechanisms with metaplasticity. Such persistent inhibition of long-term potentiation and facilitation of long-term depression might lead to activity-dependent synapse weakening and contribute to cognitive impairments. PMID- 18395201 TI - Diabetes and other vascular risk factors for dementia: which factor matters most? A systematic review. AB - Vascular risk factors, such as type 2 diabetes, hypertension, obesity and dyslipidaemia often co-occur. Each of these factors has been associated with an increased risk of dementia, but it is uncertain which factor imposes the greatest risk. Moreover, the effect of age at time of exposure may differ across factors. This paper systematically reviews the evidence for the association of each of these risk factors with dementia. Longitudinal population-based studies that assessed the incidence of dementia in relation to diabetes (n=14), hypertension (n=13), dyslipidaemia (n=8) or obesity (n=9) were included. All four risk factors were indeed associated with an increased risk of dementia, but the results of studies on diabetes and obesity were most consistent. The magnitude of the effects was comparable across the risk factors, with odds ratios for 'any dementia' around 1.5. For hypertension, obesity and dyslipidaemia age appeared to modulate the association: the risk of dementia was generally largest in studies that measured the risk factor in midlife (compared to late life) and had a long follow-up time. At midlife, the population attributable risk of dementia was highest for hypertension, up to 30% of cases of late life dementia. Later in life diabetes appears to convey the highest risk of dementia. This review shows that vascular risk factors should be regarded as a major target for preventive measures, but that timing of such measures appears to be critical. PMID- 18395202 TI - Pax3 activation promotes the differentiation of mesenchymal stem cells toward the myogenic lineage. AB - Mesenchymal stem cells (MSCs) residing within the bone marrow (BM) differentiate into multiple lineages, including fat, bone, and cartilage. Because MSCs are multipotent and have a great capacity to be expanded in vitro, these cells are an attractive candidate for clinical applications to repair or regenerate damaged tissues of mesenchymal origin. However, application of MSCs to muscle degenerative diseases has been hampered by the poor differentiation of MSCs into the muscle lineage. To date most methods require the presence of strong non physiological agents, such as azacytidine. In the present study we explored the potential of Pax3, the master regulator of the embryonic myogenic program, to promote myogenic differentiation from MSCs. Our results clearly demonstrate that Pax3 promotes the differentiation of MSCs towards the myogenic lineage, which occurs at the expense of other mesenchymal lineages including fat, bone, and cartilage. This effect is cell type-selective since Pax3 overexpression in endothelial cells fails to promote myogenesis. These results highlight the potential of regulating transcriptional pathways to direct differentiation of adult stem cells. PMID- 18395203 TI - Eimeria tenella: analysis of differentially expressed genes in the monensin- and maduramicin-resistant lines using cDNA array. AB - Drug resistance in coccidial populations has been a major problem to the prophylactic chemotherapy. Nevertheless, the mechanisms of the resistance are still poorly understood. In this report, cDNA array was designed based on the cDNA library for analysis of gene expression profile of the drug-resistant lines and their sensitive parental lines of Eimeria tenella. Two thousand eight hundred and six ESTs (expressed sequence tags) were obtained from 9600 clones which were randomly derived from the cDNA library with the 3' end sequencing. A total of 1424 TUTs (tentative unique transcripts) were determined from the database of our ESTs by bioinformatics analysis, from which a cDNA array was developed. The comparison of monensin-resistant line (MonR) and maduramicin-resistant line (MadR) with their sensitive parental lines was undertaken independently. It was observed that the number of the up-regulated genes was 5.58-fold more than that of the down-regulated genes in MonR when compared with its parental line. The up regulated genes were mainly involved in cytoskeletal rearrangements and energy metabolism. In MadR, the number of the down-regulated genes was 3.07-fold more than the up-regulated genes, which were mainly related to invasion and cytoskeletal genes. However, in MadR the level of the glycometabolism-related and potential transporter genes were reduced. Our data suggest that the mechanisms of monensin and maduramicin resistance of E. tenella might be a very complex process. PMID- 18395204 TI - Gender-specific expression of multiple estrogen receptors, growth hormone receptors, insulin-like growth factors and vitellogenins, and effects of 17 beta estradiol in the male tilapia (Oreochromis mossambicus). AB - Gender-specific expression of estrogen receptors (ER alpha and ER beta), growth hormone receptors (GHR1 and GHR2), insulin-like growth factors (IGF-I and IGF-II) and three vitellogenins (Vgs A-C) was examined in the liver, gonad, pituitary, and brain of sexually mature male, female, and 17 beta-estradiol (E2)-treated male tilapia (Oreochromis mossambicus). Reflecting greater growth rate in male tilapia, hepatic expression of GHR1, GHR2, IGF-I and IGF-II as well as plasma IGF I levels were higher in males than in females, whereas the expression of Vgs A-C and ER alpha was higher in females. On the other hand, expression of all genes measured was higher in the ovary than in testis. Forty eight hours after E2 injection (5 microg/g) into male fish, hepatic expression of most transcripts measured were altered to levels that were similar to those seen in females. The changes included decreased expression of GHR1, GHR2, IGF-I, and IGF-II, and increased expression of ER alpha and Vgs A-C. E2 treatment also increased Vg and decreased IGF-I in the plasma. Brain expression of ER alpha, ER beta, GHR1, and IGF-I was higher in females than in males, whereas pituitary expression of GHR2 and IGF-I was lower in females; only brain expression of GHR1 was increased by E2 treatment. These findings suggest that E2 stimulates Vg production primarily through activation of ER alpha and down-regulation of the GH/IGF-I axis, thus shifting energy from somatic growth towards vitellogenesis at the level of the liver. PMID- 18395205 TI - Ovarian P450 aromatase activity in the catfish Heteropneustes fossilis: seasonal changes and effects of catecholestrogens. AB - Ovarian microsomal aromatase (P450arom) activity was studied in relation to season and incubation of follicles with catecholestrogens [(2-hydroxyestradiol 17beta (2-OHE2) and 2-methoxyestradiol-17 beta (2-methoxyE2)] using a product (estradiol-17 beta) assay. Peak P450arom activity was noticed in late preparatory phase (April) and it decreased significantly in pre-spawning, spawning and post spawning phases to give the lowest value in resting phase. Apparent Km and Vmax of the enzyme varied significantly and the values were high in the preparatory (vitellogenic) phase (Km 74.62+/-1.73 nM, Vmax 0.81+/-0.01 pmol/mg protein/min) and low in the spawning (post-vitellogenic) phase (Km 62.01+/-1.68 nM, Vmax 0.69+/-0.002 pmol/mg protein/min). The incubation of the ovarian microsomes with 2-OHE2 elicited significant biphasic effects on enzyme activity. In the vitellogenic phase, concentrations of the steroid up to 1 microM inhibited enzyme activity significantly with the highest inhibition at 10nM. However, in the post vitellogenic ovary, the highest inhibition was registered at 100 nM. The higher concentrations (10 microM or 100 microM) did not elicit any significant change compared to the control groups. A comparison of the aromatase inhibition index (AI50, indicates 50% inhibition of aromatase activity) of fadrozole, a known aromatase inhibitor and 2-OHE2 shows that the AI50 was 4.4 nM for fadrozole and 0.864 nM (vitellogenic phase) and 1.31 nM (post-vitellogenic phase) for 2-OHE2 indicating higher potency of the latter. The incubation of the ovarian microsomes with 2-methoxyE2 increased enzyme activity only at the higher concentrations (1 100 microM). The results show seasonality in the potential of the ovary to synthesize E2 and the potent enzyme inhibiting activity of 2-OHE2, which is reported for the first time. PMID- 18395207 TI - Frequent recombination events generate diversity within the multi-copy variant antigen gene families of Plasmodium falciparum. AB - The human malaria parasite Plasmodium falciparum utilises a mechanism of antigenic variation to avoid the antibody response of its human host and thereby generates a long-term, persistent infection. This process predominantly results from systematic changes in expression of the primary erythrocyte surface antigen, a parasite-produced protein called PfEMP1 that is encoded by a repertoire of over 60 var genes in the P. falciparum genome. var genes exhibit extensive sequence diversity, both within a single parasite's genome as well as between different parasite isolates, and thus provide a large repertoire of antigenic determinants to be alternately displayed over the course of an infection. Whilst significant work has recently been published documenting the extreme level of diversity displayed by var genes found in natural parasite populations, little work has been done regarding the mechanisms that lead to sequence diversification and heterogeneity within var genes. In the course of producing transgenic lines from the original NF54 parasite isolate, we cloned and characterised a parasite line, termed E5, which is closely related to but distinct from 3D7, the parasite used for the P. falciparum genome nucleotide sequencing project. Analysis of the E5 var gene repertoire, as well as that of the surrounding rif and stevor multi-copy gene families, identified examples of frequent recombination events within these gene families, including an example of a duplicative transposition which indicates that recombination events play a significant role in the generation of diversity within the antigen encoding genes of P. falciparum. PMID- 18395206 TI - AR, apoE, and cognitive function. AB - Reduced androgen levels in aged men and women might be risk factors for age related cognitive decline and Alzheimer's disease (AD). Ongoing clinical trials are designed to evaluate the potential benefit of estrogen in women and of testosterone in men. In this review, we discuss the potential beneficial effects of androgens and androgen receptors (ARs) in males and females. In addition, we discuss the hypothesis that AR interacts with apolipoprotein (apoE)4, encoded by epsilon4 and a risk factor for age-related cognitive decline and AD, and the potential consequences of this interaction. PMID- 18395208 TI - Contemporary risk factor control and walking dysfunction in individuals with peripheral arterial disease: NHANES 1999-2004. AB - BACKGROUND: Lower extremity peripheral arterial disease (PAD) is a coronary heart disease (CHD) risk equivalent. Selected studies have demonstrated less intense risk factor management and diminished mobility in individuals with PAD as compared to individuals with clinical recognized CHD. However, comparable data have not been reported from a nationally representative population. OBJECTIVES: To assess the prevalence, treatment, and control of cardiovascular risk factors among individuals with PAD as defined by an ankle-brachial index (ABI) <0.90 (but without recognized CHD) as compared with individuals with recognized CHD (but without PAD). A second objective was to evaluate the diagnostic accuracy of measures of walking dysfunction to identify individuals with PAD. DESIGN, SETTING, AND PARTICIPANTS: We analyzed data from 7571 participants aged 40 or older who participated in the National Health and Nutrition Examination Survey (NHANES) 1999-2004, a nationally representative cross-sectional survey of the U.S. population. RESULTS: The prevalence of PAD without CHD was 4.1% (95% CI 3.6 4.5) compared to 7.9% (7.1, 8.9) for CHD (without PAD). Hypertension prevalence was similar, but treatment and control rates were lower among individuals with PAD compared to CHD (treatment: 69% vs. 84%, p<0.001; control: 50% vs. 63%, p=0.01). Treatment of hypercholesterolemia was lower among individuals with PAD (54% vs. 79%, p<0.001) but control was similar (83% vs. 85%, p=0.78). Diabetes awareness, treatment, and control did not differ between the two groups. Walking mobility limitations were specific, but insensitive, for the identification of individuals with PAD. CONCLUSION: PAD in the absence of clinically recognized CHD is under-treated and poorly controlled in the general U.S. population. Leg symptoms are not adequate to identify individuals with PAD, who are at high risk of ischemic events. PMID- 18395209 TI - Deficits in the way to achieve balance related to mechanisms of dynamic stability control in the elderly. AB - The purpose of this study was to examine the postural corrections related to components of dynamic stability aimed to increase our understanding of successful postural control among the elderly population. This was done by comparing balance behaviour of older adults who were able to recover stability (stable) and others who failed to regain stability (unstable) with a single step after a forward fall. Thirty-eight old male adults (64+/-3yr, 176+/-6cm, 78.5+/-7.8kg) had to recover balance after a sudden induced forward fall. All participants performed maximal isometric ankle plantarflexion and knee extension contractions on a dynamometer. The elongation of the gastrocnemius medialis and the vastus lateralis tendon and aponeuroses during isometric contraction was examined by ultrasonography. There were no differences in leg-extensor muscle strength or tendon stiffness between the two groups showing that the muscle tendon capacities may not be the reason for the observed differences in dynamic stability control. The unstable participants created a higher horizontal ground reaction push-off force of the support limb in the second part ( approximately 260ms after release) of the phase until touchdown leading to an unstable body position at touchdown. The results indicate deficits in the way to achieve balance related to mechanisms responsible for dynamic stability control within the elderly population. PMID- 18395210 TI - Reliability of upper and lower extremity anthropometric measurements and the effect on tissue mass predictions. AB - Accurate modeling of soft tissue motion effects relative to bone during impact requires knowledge of the mass of soft and rigid tissues in living people. Holmes et al., [2005. Predicting in vivo soft tissue masses of the lower extremity using segment anthropometric measures and DXA. Journal of Applied Biomechanics, 21, 371 382] developed and validated regression equations to predict the individual tissue masses of lower extremity segments of young healthy adults, based on simple anthropometric measurements. However, the reliability of these measurements and the effect on predicted tissue mass estimates from the equations has yet to be determined. In the current study, two measurers were responsible for collecting two sets of unilateral measurements (25 male and 25 female subjects) for the right upper and lower extremities. These included 6 lengths, 6 circumferences, 8 breadths, and 4 skinfold thicknesses. Significant differences were found between measurers and between sexes, but these differences were relatively small in general (75-80% of between-measurer differences were <1cm). Within-measurer measurement differences were smaller and more consistent than those between measurers in most cases. Good to excellent reliability was demonstrated for all measurement types, with intra-class correlation coefficients of 0.79, 0.86, 0.85 and 0.86 for lengths, circumferences, breadth and skinfolds, respectively. Predicted tissue mass magnitudes were moderately affected by the measurement differences. The maximum mean errors between measurers ranged from 3.2% to 24.2% for bone mineral content and fat mass, for the leg and foot, and the leg segments, respectively. PMID- 18395211 TI - Dynamic gait stability index based on plantar pressures and fuzzy logic. AB - Stability during locomotion, or dynamic stability, is critical to ensure safe locomotion and a high quality of life. A dynamic stability measure should be easily applied in a clinical setting and must provide a quantitative index that can be used for comparisons over a range of tasks and environments. Plantar foot pressure data acquired by shoe-insole sensors have potential to provide such a measure. To generate a quantitative dynamic gait stability index, six gait parameters were extracted from a commercial plantar pressure measurement system (F-Scan): anterior-posterior (A/P) center of force (CoF) motion, medial-lateral (M/L) CoF motion, maximum lateral position, cell triggering, stride time (ST), and double support time (DST). A fuzzy logic controller combined these six parameters and generated the index. To validate the stability index, 15 healthy subjects performed four tasks intended to induce increasing levels of instability. Fifty-seven gait parameter combinations were assessed to determine the most effective index. A combination of A/P motion, M/L motion, maximum lateral position, and cell triggering parameters was the most consistently effective index across all subjects. However, small changes in ST and DST for able-bodied subjects may have reduced the effectiveness of these measures in the index calculation. The index combining all six parameters should be investigated further with populations with disabilities or pathological gait. PMID- 18395212 TI - Structural simulations of prosthetic tri-leaflet aortic heart valves. AB - This study presents a combined computational and experimental approach for the nonlinear structural simulations of polymeric tri-leaflet aortic valves (PAVs). Nonlinear shell-based and quasi-static finite-element (FE) structural models are generated for a prosthetic valve geometry that includes the leaflets, stents and root materials, such as the bottom base and outside walls. The PAV structural model is subject to an ensemble averaged transvalvular pressure waveform measured from repeated in vitro tests conducted with a left heart simulator. High resolution optical measurements are used to measure the in vitro kinematics of the leaflets and the stents. Qualitative and quantitative deformation measures are defined in order to compare the predicted kinematics from the PAV models with the in vitro measurements. Six new quantitative deformation metrics are introduced. They include three distances measuring the current PAV geometric center to the leaflet edges while additional three distances define the stent post-to-stent post (SPTSP) distances. The structural model is able to predict the kinematic deformation metrics with maximum errors around 10% especially in systole where the displacements are larger in magnitude. The combined structural modeling with experimental simulations along with the new proposed deformation metrics provide an effective way to study the PAV structural behavior and a path for improving the structural design of prosthetic valves. PMID- 18395213 TI - A theoretical analysis of an optimal chainring shape to maximize crank power during isokinetic pedaling. AB - Previous studies have sought to improve cycling performance by altering various aspects of the pedaling motion using novel crank-pedal mechanisms and non circular chainrings. However, most designs have been based on empirical data and very few have provided significant improvements in cycling performance. The purpose of this study was to use a theoretical framework that included a detailed musculoskeletal model driven by individual muscle actuators, forward dynamic simulations and design optimization to determine if cycling performance (i.e., maximal power output) could be improved by optimizing the chainring shape to maximize average crank power during isokinetic pedaling conditions. The optimization identified a consistent non-circular chainring shape at pedaling rates of 60, 90 and 120 rpm with an average eccentricity of 1.29 that increased crank power by an average of 2.9% compared to a conventional circular chainring. The increase in average crank power was the result of the optimal chainrings slowing down the crank velocity during the downstroke (power phase) to allow muscles to generate power longer and produce more external work. The data also showed that chainrings with higher eccentricity increased negative muscle work following the power phase due to muscle activation-deactivation dynamics. Thus, the chainring shape that maximized average crank power balanced these competing demands by providing enough eccentricity to increase the external work generated by muscles during the power phase while minimizing negative work during the subsequent recovery phase. PMID- 18395214 TI - Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels. AB - Molecular imprinting is a method for making selective binding sites in synthetic polymers using a molecular template. The aim of this study is to prepare lysozyme imprinted supermacroporous cryogels which can be used for the purification of lysozyme (Lyz) from egg white. N-Methacryloyl-(L)-histidinemethylester (MAH) was chosen as the metal-coordinating monomer. In the first step, Cu2+ was complexed with MAH and the lysozyme-imprinted poly(HEMA-MAH) [Lyz-MIP] cryogel were produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) in an ice bath. After that, the template (i.e., lysozyme) was removed using 0.05 M phosphate buffer containing 1M NaCl (pH 8.0). The maximum lysozyme adsorption capacity was 22.9 mg/g polymer. The relative selectivity coefficients of Lyz-MIP cryogel for lysozyme/bovine serum albumin and lysozyme/cytochrome c were 4.6 and 3.2 times greater than non-imprinted poly(HEMA MAH) (NIP) cryogel, respectively. Purification of lysozyme from egg white was also monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purity of the desorbed lysozyme was about 94% with recovery about 86%. The Lyz-MIP cryogel could be used many times without decreasing the adsorption capacity significantly. PMID- 18395215 TI - Quaternary size distribution of soluble aggregates of glutathione-S-transferase purified viral protein as determined by asymmetrical flow field flow fractionation and dynamic light scattering. AB - Polyomavirus VP1 protein in pentamer form was expressed in E. coli and purified using glutathione-S-transferase (GST) affinity chromatography. Purified GST tagged protein was found to exist as soluble aggregates with a size distribution of 1-52 tagged pentamers (340-1800 x 10(3)kDa), as determined by asymmetrical flow field flow fractionation with multiple angle light scattering (AFFFF-MALS). Aggregation did not inhibit tag removal by enzymatic cleavage, implying that the quaternary structure of the VP1 pentamers had been maintained. Elution gel filtration (EGF) was utilized to prepare a solution enriched with protein small enough to access resin pores (LMWe) as well as solution enriched with protein excluded from resin pores (HMWe). Material size distributions within both solutions were determined using AFFFF-MALS (radius of gyration LMWe: 5-10nm; HMWe: 10-35 nm) and dynamic light scattering (DLS) (hydrodynamic diameter LMWe: 10-90 nm; HMWe: 20-300 nm). DLS and AFFFF-MALS analysis of each fraction of affinity chromatography purified material identified the elution profiles of large and small aggregate structures. DLS readings of all fractions were significantly affected by the presence of high molecular weight aggregates, with Z-average hydrodynamic diameter values reflecting the mass ratio of large and small aggregate structures in a solution. The methods utilized in this study have the potential to be used during chromatographic purification of all proteins that exist as soluble aggregates to determine size distribution. The finding that GST tagged viral proteins exist as soluble aggregates has implications for existing immunological studies that utilize them. PMID- 18395217 TI - Seedless synthesis of octahedral gold nanoparticles in condensed surfactant phase. AB - We report a seedless synthetic method of gold octahedral nanoparticles in an aqueous phase. Eight facets with {111} crystalline structures of octahedral nanoparticles could be formed in an aqueous medium when the gold salt was reduced by ascorbic acid at room temperature in the presence of cetyltrimethylammonium bromide as a shape-inducing agent, and hydrogen peroxide as a reaction promoter. The growth kinetics and surface crystalline structures were characterized by UV vis spectroscopy, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy. PMID- 18395218 TI - Microviscosity of hydrophobically modified hydroxyethyl cellulose aqueous solutions. AB - The microviscosity of hydrophobically modified hydroxyethyl cellulose (HMHEC) aqueous solutions is experimentally determined by conductometry with added ions as probe. Compared to its bulk viscosity, the microviscosity of HMHEC solution could be lower by four orders of magnitude. Since the electric conductivity reduction of added NaCl is almost the same for HMHEC and its unmodified counterpart at an equal weight concentration, one can conclude that the hydrophobic modification for the polymer hardly has any effect on the solution's microviscosity. PMID- 18395219 TI - Comparison of performance on two nonverbal intelligence tests by adolescents with and without language impairment. AB - Definitions of specific language impairment (SLI), for both research and clinical purposes, often state that nonverbal IQ scores must be within normal limits. This use of nonverbal IQ has been criticized on several grounds, including lack of equivalence between tests. In the current study, a sample of 204 adolescents with and without language impairment took two nonverbal IQ tests. Scores on the tests were compared, as was the classification of individuals into diagnostic categories according to the two tests. Results indicated that there were significant differences between test scores for clinical subgroups, and classification of individuals as having SLI varied substantially depending on the test used. The findings are interpreted as providing further evidence that nonverbal IQ should be used only cautiously, if at all, in defining SLI. LEARNING OUTCOMES: Readers will (1) understand problems with the use of nonverbal IQ criteria in diagnosis of SLI and (2) be able to cite data showing that different nonverbal IQ tests can lead to different diagnoses. PMID- 18395220 TI - Real time detection of anthrax spores using highly specific anti-EA1 recombinant antibodies produced by competitive panning. AB - We describe a targeted approach for the production of biological recognition elements capable of fast, specific detection of anthrax spores on biosensor surfaces. The aim was to produce single chain antibodies (scFvs) to EA1, a Bacillus anthracis S-layer protein that is also present, although not identical, in related to Bacillus species. The aim of the work was to produce antibodies that would detect B. anthracis EA1 protein and intact spores with a high degree of specificity, but would not detect other Bacillus species. Existing monoclonal antibodies were evaluated and found to recognise B. anthracis EA1 and S-layer proteins from other closely related Bacillus species. Recombinant anti-EA1 scFvs were isolated from B. anthracis immune library that contained antibody genes raised against B. anthracis spores and purified exosporium. Two approaches for scFv selection were used; standard (non-competitive) panning, and competitive panning. The non-competitive biopanning strategy isolated scFvs that recognised EA1 from B. anthracis, but also cross-reacted with other Bacillus species. In contrast, the competitive panning approach used S-layer proteins from other Bacillus species to generate scFvs that were highly specific to B. anthracis EA1 and demonstrated apparent nanomolar binding affinities. Specific, real time detection of B. anthracis spores was demonstrated with these scFvs using an evanescent wave biosensor, the Resonant Mirror. The approach described can be used to generate specific antibodies to any desired target where homologous proteins also exist in closely related species, and demonstrates clear advantages to using recombinant technology to produce biological recognition elements for detection of biological threat agents. PMID- 18395221 TI - Enumeration of functionally active anti-Aspergillus T-cells in human peripheral blood. AB - Invasive aspergillosis remains a life-threatening complication in patients undergoing allogeneic stem cell transplantation (SCT). Since CD4(+) T-cells provide a critical secondary defense against Aspergillus spp., the quantification of "functional" anti-Aspergillus T-cells might be important in the clinical care of allogeneic transplant patients. We present a rapid, simple and reproducible method to enumerate functionally active, cytokine-producing anti-Aspergillus T cells in peripheral blood by means of flow cytometry, by which these cells were also phenotypically characterized as memory CD4(+) T-cells. When using 100,000 PBMCs and requiring a minimum of 50 events, at least one anti-Aspergillus T-cell among 1000 CD4(+) T-cells can be detected. Compared to healthy individuals, the number of anti-Aspergillus T-cells in patients up to one year after SCT was significantly lower. The presented method might help to define hematopoietic transplant recipients who will benefit from adoptive transfer of anti-Aspergillus T cells. PMID- 18395222 TI - Insecticidal activity of a basement membrane-degrading protease against Heliothis virescens (Fabricius) and Acyrthosiphon pisum (Harris). AB - ScathL is a cathepsin L-like cysteine protease derived from the flesh fly Sarcophaga peregrina that functions in basement membrane (BM) remodeling during insect development. A recombinant baculovirus expressing ScathL (AcMLF9.ScathL) kills larvae of the tobacco budworm, Heliothis virescens, significantly faster than the wild-type virus. Here, we show that the occurrence of larval melanization prior to death was closely associated with the onset of high cysteine protease activity of ScathL in the hemolymph of fifth instars infected with AcMLF9.ScathL, but not with AcMLF9.ScathL.C146A, a recombinant baculovirus expressing a catalytic site mutant of ScathL. Fragmented fat body, ruptured gut and malpighian tubules, and melanized tracheae were observed in AcMLF9.ScathL infected larvae. Phenoloxidase activity in hemolymph was unchanged, but the pool of prophenoloxidase was significantly reduced in virus-infected larvae and further reduced in AcMLF9.ScathL-infected larvae. The median lethal dose (LD(50)) for purified ScathL injected into fifth-instar H. virescens was 11.0 microg/larva. ScathL was also lethal to adult pea aphids, Acyrthosiphon pisum with a similar loss of integrity of the gut and fat body. Injection with purified ScathL.C146A or bovine trypsin at 20 microg/larva did not produce any effect in either insect. These results illustrate the potent insecticidal effects of ScathL cysteine protease activity and the potential for use of ScathL in development of insect resistant transgenic plants when combined with an appropriate delivery system. PMID- 18395223 TI - Crystal structure and Raman studies of dsFP483, a cyan fluorescent protein from Discosoma striata. AB - To better understand the diverse mechanisms of spectral tuning operational in fluorescent proteins (FPs), we determined the 2.1-A X-ray structure of dsFP483 from the reef-building coral Discosoma. This protein is a member of the cyan class of Anthozoa FPs and exhibits broad, double-humped excitation and absorbance bands, with a maximum at 437-440 nm and a shoulder at 453 nm. Although these features support a heterogeneous ground state for the protein-intrinsic chromophore, peak fluorescence occurs at 483 nm for all excitation wavelengths, suggesting a common emissive state. Optical properties are insensitive to changes in pH over the entire range of protein stability. The refined crystal structure of the biological tetramer (space group C2) demonstrates that all protomers bear a cis-coplanar chromophore chemically identical with that in green fluorescent protein (GFP). To test the roles of specific residues in color modulation, we investigated the optical properties of the H163Q and K70M variants. Although absorbance bands remain broad, peak excitation maxima are red shifted to 455 and 460 nm, emitting cyan light and green light, respectively. To probe chromophore ground-state features, we collected Raman spectra using 752-nm excitation. Surprisingly, the positions of key Raman bands of wild-type dsFP483 are most similar to those of the neutral GFP chromophore, whereas the K70M spectra are more closely aligned with the anionic form. The Raman data provide further evidence of a mixed ground state with chromophore populations that are modulated by mutation. Possible internal protonation equilibria, structural heterogeneity in the binding sites, and excited-state proton transfer mechanisms are discussed. Structural alignments of dsFP483 with the homologs DsRed, amFP486, and zFP538 K66M suggest that natural selection for cyan is an exquisitely fine-tuned and highly cooperative process involving a network of electrostatic interactions that may vary substantially in composition and arrangement. PMID- 18395224 TI - The controlling roles of Trp60 and Trp95 in beta2-microglobulin function, folding and amyloid aggregation properties. AB - Amyloidosis associated to hemodialysis is caused by persistently high beta(2) microglobulin (beta(2)m) serum levels. beta(2)m is an intrinsically amyloidogenic protein whose capacity to assemble into amyloid fibrils in vitro and in vivo is concentration dependent; no beta(2)m genetic variant is known in the human population. We investigated the roles of two evolutionary conserved Trp residues in relation to beta(2)m structure, function and folding/misfolding by means of a combined biophysical and functional approach. We show that Trp60 plays a functional role in promoting the association of beta(2)m in class I major histocompatibility complex; it is exposed to the solvent at the apex of a protein loop in order to accomplish such function. The Trp60-->Gly mutation has a threefold effect: it stabilizes beta(2)m, inhibits beta(2)m amyloidogenic propensity and weakens the interaction with the class I major histocompatibility complex heavy chain. On the contrary, Trp95 is buried in the beta(2)m core; the Trp95-->Gly mutation destabilizes the protein, which is unfolded in solution, yielding nonfibrillar beta(2)m aggregates. Trp60 and Trp95 therefore play differential and complementary roles in beta(2)m, being relevant for function (Trp60) and for maintenance of a properly folded structure (Trp95) while affecting in distinct ways the intrinsic propensity of wild-type beta(2)m towards self-aggregation into amyloid fibrils. PMID- 18395225 TI - Sequence and structural determinants of strand swapping in cadherin domains: do all cadherins bind through the same adhesive interface? AB - Cadherins are cell surface adhesion proteins important for tissue development and integrity. Type I and type II, or classical, cadherins form adhesive dimers via an interface formed through the exchange, or "swapping", of the N-terminal beta strands from their membrane-distal EC1 domains. Here, we ask which sequence and structural features in EC1 domains are responsible for beta-strand swapping and whether members of other cadherin families form similar strand-swapped binding interfaces. We created a comprehensive database of multiple alignments of each type of cadherin domain. We used the known three-dimensional structures of classical cadherins to identify conserved positions in multiple sequence alignments that appear to be crucial determinants of the cadherin domain structure. We identified features that are unique to EC1 domains. On the basis of our analysis, we conclude that all cadherin domains have very similar overall folds but, with the exception of classical and desmosomal cadherin EC1 domains, most of them do not appear to bind through a strand-swapping mechanism. Thus, non classical cadherins that function in adhesion are likely to use different protein protein interaction interfaces. Our results have implications for the evolution of molecular mechanisms of cadherin-mediated adhesion in vertebrates. PMID- 18395226 TI - Identification of a new homozygous frameshift insertion mutation in the SIL1 gene in 3 Japanese patients with Marinesco-Sjogren syndrome. AB - Marinesco-Sjogren syndrome (MSS) is an autosomal recessive multisystem disorder characterized by cerebellar ataxia, cataracts, progressive muscular weakness, and developmental and mental retardation. Recently, mutations in the SIL1 gene on chromosome 5q31 have been shown to be a cause of MSS. We sequenced the entire SIL1-coding region in 3 unrelated Japanese patients with classical MSS and identified a novel homozygous frameshift insertion mutation, 936_937insG, in exon 9 in all 3 patients. PMID- 18395227 TI - A protein interaction network associated with asthma. AB - Identifying candidate genes related to complex diseases or traits and mapping their relationships require a system-level analysis at a cellular scale. The objective of the present study is to systematically analyze the complex effects of interrelated genes and provide a framework for revealing their relationships in association with a specific disease (asthma in this case). We observed that protein-protein interaction (PPI) networks associated with asthma have a power law connectivity distribution as many other biological networks have. The hub nodes and skeleton substructure of the result network are consistent with the prior knowledge about asthma pathways, and also suggest unknown candidate target genes associated with asthma, including GNB2L1, BRCA1, CBL, and VAV1. In particular, GNB2L1 appears to play a very important role in the asthma network through frequent interactions with key proteins in cellular signaling. This network-based approach represents an alternative method for analyzing the complex effects of candidate genes associated with complex diseases and suggesting a list of gene drug targets. The full list of genes and the analysis details are available in the following online supplementary materials: http://biosoft.kaist.ac.kr:8080/resources/asthma_ppi. PMID- 18395228 TI - Ecotoxicological evaluation of polycyclic aromatic hydrocarbons using marine invertebrate embryo-larval bioassays. AB - The toxicity of polycyclic aromatic hydrocarbons (PAHs) was determined using mussel, sea-urchin and ascidian embryo-larval bioassays. Fluorescent light exposure enhanced phenanthrene, fluoranthene, pyrene and hydroxypyrene toxicity in comparison with dark conditions, but not naphthalene and fluorene toxicity. The toxicity of PAHs was inversely related to their K(OW) values following QSAR models derived for baseline toxicity of general narcotics, whereas the obtained regression using toxicity data from photoactivated PAHs significantly departed from the general narcosis model. Also, the mixture toxicity of five PAHs to the larval growth of the sea-urchin was compared with predictions derived from the concentration addition concept, indicating less than additive effects. Finally, we compared our toxicity data with worst-case environmental concentrations in order to provide a preliminary estimate of the risk to the marine environment. Naphthalene, fluorene and pyrene are not considered to pose a risk to sea-urchin, mussel or ascidian larvae, whilst phenanthrene and fluoranthene may pose a risk for mussel and sea-urchin. Moreover, a higher risk for those species is expected when we consider the photoactivation of the PAHs. PMID- 18395229 TI - Regional variations in trace element concentrations in tissues of black tiger shrimp Penaeus monodon (Decapoda: Penaeidae) from South Vietnam. AB - The goal of the present study was to examine the specific bioaccumulation of 22 trace elements in muscle, exoskeleton and hepatopancreas of black tiger shrimp Penaeus monodon from the Mekong River Delta (MRD), and the South Key Economic Zone (SKEZ), South Vietnam. The general tendency in most trace element concentrations among different tissues were hepatopancreas>exoskeleton>muscle. Comparisons of trace element levels in tissues between the two regions showed that concentrations of Se in muscle and As in all three tissues were higher in SKEZ; whereas in MRD, the higher concentrations of most elements such as Mn, Cu, Cd, Ba, Hg, were observed in tissues. These geographical variations in trace element levels may reflect the differences in human activities between the two regions of South Vietnam. The target hazard quotient (THQ) values for trace elements (<1) indicate that local residents are not exposed to potential health risks via the consumption of shrimp. PMID- 18395230 TI - On global and local critical points of extended contact process on homogeneous trees. AB - We study spatial stochastic epidemic models called households models. The households models have more than two states at each vertex of a graph in contrast to the contact process. We show that, in the households models on trees, two thresholds of infection rates characterize epidemics. The global critical infection rate is defined by epidemic occurrence. However, some households may be eventually disease-free even for infection rates above the global critical infection rate, in as far as they are smaller than the local critical point. Whether the global one is smaller than the local one depends on the graph and the model. We show that, in the households models, the global one is smaller than the local one on homogeneous trees. PMID- 18395231 TI - [Giant prebulbomedullary arachnoid cyst. Case report and review of the literature]. AB - Usual locations of arachnoid cyst are the middle cranial fossa in 50-60%, cerebellopontine angle (10%) and suprasellar area (10%). Most of these malformations are asymptomatic. Premedullar arachnoid cysts are extremely rare. All previous cases reported were operated. We report a case of an asymptomatic giant craniocervical junction arachnoid cyst with a follow up of five years. In 2002, an adolescent consulted for persistent cervical pain. Encephalic MR showed a giant ventral craniocervical junction arachnoid cyst. Neurologic examination was normal. Conservative treatment was decided with a clinical follow up and repeated MR in case of persistent cervicalgia. Craniocervical junction arachnoid cysts are anecdotic. Medical care cannot be standardized. Pathogenesis and management are discussed. PMID- 18395233 TI - Neuropsychological correlates of whole brain atrophy in Alzheimer's disease. AB - Alzheimer's disease (AD) is associated with excess whole brain volume loss, and progressive cognitive impairment. We aimed to study the extent to which these two potential biomarkers of AD progression are correlated. Forty-six patients with sporadic AD were tested with a neuropsychometric battery including test of verbal and visual memory, vocabulary, arithmetic, naming, visuoperceptual skills and reasoning at two time-points, approximately 1 year apart; annualised rates of change for each test were calculated. Each subject also attended for up to twelve T1-weighted volumetric MRI scans at fixed intervals over a 2-year period. For each individual all possible scan-pairs were positionally registered, and whole brain atrophy rates were calculated using the brain boundary shift integral. Linear mixed models were used to investigate associations between atrophy rate and coincident change in each neuropsychometric score. Each model estimated the effect of a unit change in score, plus the additional effect of a fall to floor, after adjusting for baseline levels. 467 MRI scans were performed, permitting 2199 individual measures of change to be made. The model-derived mean atrophy rate was 2.23% per year with a between-subject SD of 0.99% per year. Increasing atrophy rate was significantly associated with rate of change in a number of non memory based neuropsychological scores, with the strongest association seen with longitudinal change in matrix reasoning (p=0.004). These results provide further evidence that cerebral atrophy is a clinically relevant marker of AD progression. This methodology whereby data from patients falling to floor on a given test may be included and accounted for, rather than discarded, may find broader application in clinical studies incorporating neuropsychometric outcomes. PMID- 18395234 TI - Finding the hidden faces: schizophrenic patients fare worse than healthy subjects. AB - Humans have a special ability to recognize human faces that transcends and is separate from the usual discrimination abilities of the visual system. Schizophrenia patients are known to have an impaired ability to recognize facial affect, a deficit that may stem from a more profound problem of face identification and perception. The special skill of detecting human faces relies upon numerous capabilities, including gestalt perception, "filling-in", and proper gaze scanning, facilities that recent research has shown to be deficient in schizophrenia patients. We therefore hypothesized that schizophrenia patients have a deficit in their perception and structural analysis of human faces that will manifest in their difficulty in detecting faces hidden within neutral pictures. We tested the ability of 35 schizophrenia patients and 32 healthy controls at detecting hidden human faces and animals or animal faces hidden within eight pictures. Overall, the patients recognized fewer items compared to controls, however, it was specifically the human faces in which this difference was significant, suggesting that schizophrenic patients have a specific deficit in their human face recognition system. PMID- 18395235 TI - Yield of fluorescence microscopy versus culture for tuberculosis at a middle income country referral hospital. AB - The aim of this study was to determine the usefulness of fluorescence microscopy (FM) at a referral centre in a middle-income country. Direct Ziehl-Neelsen (ZN) and direct, as well as concentrated, smear FM were performed on 2179 suspect sputa, with Lowenstein-Jensen (LJ) culture as the gold standard. ZN, direct FM and concentration FM detected 36.0, 38.6 and 37.0%, respectively, of 272 culture positive specimens. Patient-wise, there were 8.1% (126/1553) positives on any smear compared with 12.0% (187/1553) on any culture. ZN, direct FM and concentrated FM smear were positive in 43.3, 46.5 and 45.5%, respectively, of culture-proven cases. All differences between microscopy and culture were significant (P<0.001), but not those between microscopy techniques. Acid-fast bacilli (AFB) were not rare in 60% of 48 duplicate smears, positive in ZN or FM only. Simple LJ culture, but not FM on direct or concentrated smears, was thus significantly more sensitive than ZN smears. The considerable numbers of AFB found in positive direct smears from discordant microscopy sets suggest that repeating smears can improve microscopy sensitivity more than variations of correctly executed technique, provided that overload is avoided. Thus FM could be particularly useful, as it is time-saving and could protect against the sensitivity loss associated with high workload. PMID- 18395236 TI - Intestinal parasitic infections in adolescent girls from two boarding schools in southern Benin. AB - Intestinal parasitic infections (IPI), especially helminths, represent a major public health problem that increase iron deficiency anaemia in developing countries. This study investigated the prevalence, risk factors and nutritional consequences of IPIs in 180 adolescent girls aged 12-17 years living in two boarding schools in southern Benin. Data were collected using a structured questionnaire and laboratory analysis of blood and faecal samples. The relationships between socioeconomic indicators, IPIs and iron status were analysed using logistic regression analysis. Fifty percent of the subjects were infected with at least one IPI: 2% with helminths, 41% with protozoa and 7% with two or more intestinal parasites. Adolescent girls from a large family and those whose mothers were manual workers showed a higher risk of intestinal parasitism (odds ratio (OR)=3.5, 95% CI 2.5-5.2 (P=0.02) and OR=2.4, 95% CI 2.0-3.0 (P=0.03), respectively). Likewise, drinking untreated water was also a high risk factor for infection (OR=2.3, 95% CI 1.5-2.4; P=0.03). No significant association was observed between IPIs and iron deficiency or iron deficiency anaemia, which can be explained by the low wormload observed. These findings reinforce the need to involve mothers in health initiatives to control intestinal parasitism in Benin. PMID- 18395237 TI - Progesterone metabolites rapidly stimulate calcium influx in human platelets by a src-dependent pathway. AB - The effects of several steroids and their metabolites were examined for their ability to rapidly alter intracellular free calcium ([Ca(2+)](i)) in the anucleate human platelet. Earlier studies suggested that steroids had direct and rapid non-genomic effects to alter platelet physiology. The rationale for performing this study was to investigate the signal transduction events being activated by steroids. Super-physiologic concentrations (1.0-10.0microM) of beta estradiol and several estradiol metabolites and analogs potentiated (approximately twofold) the action of thrombin to elevate [Ca(2+)](i) in platelets, whereas 10.0microM progesterone inhibited the action of thrombin by 10 15%. Progesterone and beta-estradiol by themselves did not affect [Ca(2+)](i). Progesterone metabolites can achieve high blood concentrations. Some progesterone metabolites, particularly those in the beta-conformation, were potent stimulators of Ca(2+) influx and intracellular Ca(2+) mobilization in platelets. They activated phospholipase C because their ability to increase [Ca(2+)](i) was inhibited by the phospholipase C inhibitor U-73122. The ability of pregnanediol and collagen to increase [Ca(2+)](i) was inhibited by the src tyrosine kinase inhibitor PP1, whereas the actions of thrombin and thapsigargin to increase [Ca(2+)](i) were not affected by PP1. The effects of progesterone metabolites to increase [Ca(2+)](i) were observed with concentrations as low as 0.1microM. Pregnanolone synergized with thrombin to increase [Ca(2+)](i). It is hypothesized that human platelets possess receptors for progesterone metabolites. These receptors when stimulated will activate platelets by causing a rapid increase in [Ca(2+)](i). Pregnanolone, isopregnanediol and pregnanediol were the most effective stimulators of this newly identified src-dependent signal transduction system in platelets. Progesterone metabolites may regulate platelet aggregation and hence thrombosis in vivo. PMID- 18395238 TI - A subset of equine sarcoids harbours BPV-1 DNA in a complex with L1 major capsid protein. AB - Bovine papillomavirus type 1 or 2 (BPV-1, BPV-2) are accepted causal factors in equine sarcoid pathogenesis. Whereas viral genomes are consistently found and expressed within lesions, intact virions have never been detected, thus permissiveness of sarcoids for BPV-1 replication remains unclear. To reassess this issue, an immunocapture PCR (IC/PCR) was established using L1-specific antibodies to capture L1-DNA complexes followed by amplification of the viral genome. Following validation of the assay, 13 sarcoid-bearing horses were evaluated by IC/PCR. Samples were derived from 21 tumours, 4 perilesional/intact skin biopsies, and 1 serum. Tissue extracts from sarcoid-free equines served as controls. IC/PCR scored positive in 14/24 (58.3%) specimens obtained from sarcoid patients, but negative for controls. Quantitative IC/PCR demonstrated <125 immunoprecipitable viral genomes/50 microl extract for the majority of specimens. Moreover, full-length BPV-1 genomes were detected in a complex with L1 proteins. These complexes may correspond to virion precursors or intact virions. PMID- 18395239 TI - Involvement of the Escherichia coli endoribonucleases G and E in the secondary processing of RegB-cleaved transcripts of bacteriophage T4. AB - Sequence-specific endoribonuclease RegB of bacteriophage T4 cleaves early phage mRNAs and facilitates the transition between early and subsequent phases of T4 gene expression. The great majority of RegB targets have been identified in the intergenic regions of T4 transcripts, frequently in the Shine-Dalgarno sequences. Here we show that localization of RegB targets is not restricted to intergenic regions of mRNA. We detected 30 intragenic RegB sites in T4 transcripts that are differently susceptible to cleavage. Four RegB-processed mRNAs were previously shown to undergo further processing at so-called "secondary sites". We have found three additional transcripts carrying clear targets for both RegB and another endoribonuclease. We show that secondary cuts within RegB-processed T4 mRNAs are generated mainly by Escherichia coli RNase G, but that in some cases RNase E can recognize the same targets. Using plasmid-phage systems we demonstrate that T4 infection favours cleavage by the host endoribonucleases at these sites. PMID- 18395240 TI - Genetic diversity and purifying selection in West Nile virus populations are maintained during host switching. AB - To investigate differential evolutionary rates and selective forces of WNV in hosts and vectors, we measured the genetic diversity that arose during alternating passage in mosquitoes and birds. Within-host genetic diversity was monitored in each of three experimentally passed replicates, and the complete genome sequence of each WNV strain was determined after passage. The intrahost genetic diversity that arose during alternating passage was significantly greater than the diversity generated during chicken-only passage and similar to mosquito only passage. dN/dS ratios suggested purifying selection similar to chick-passed virus, but not to mosquito-passed virus. Thus, the abundant genetic variation contributed to WNV populations through infection of mosquitoes and the strong purifying selection contributed by infection of birds may be maintained despite frequent host switching. PMID- 18395241 TI - Cuttlefish camouflage: the effects of substrate contrast and size in evoking uniform, mottle or disruptive body patterns. AB - Cuttlefish are cephalopod molluscs that achieve dynamic camouflage by rapidly extracting visual information from the background and neurally implementing an appropriate skin (or body) pattern. We investigated how cuttlefish body patterning responses are influenced by contrast and spatial scale by varying the contrast and the size of checkerboard backgrounds. We found that: (1) at high contrast levels, cuttlefish body patterning depended on check size; (2) for low contrast levels, body patterning was independent of "check" size; and (3) on the same check size, cuttlefish fine-tuned the contrast and fine structure of their body patterns, in response to small contrast changes in the background. Furthermore, we developed an objective, automated method of assessing cuttlefish camouflage patterns that quantitatively differentiated the three body patterns of uniform/stipple, mottle and disruptive. This study draws attention to the key roles played by background contrast and particle size in determining an effective camouflage pattern. PMID- 18395242 TI - Dioxins, PCBs, metals, metalloids, pesticides and antimicrobial residues in wild and farmed Australian southern bluefin tuna (Thunnus maccoyii). AB - Data concerning the concentration of dioxins (PCDD/PCDFs), polychlorinated biphenyls (PCBs), metals, metalloids, pesticides and antimicrobials residues are reported for the edible portion of Australian wild and farmed southern bluefin tuna (Thunnus maccoyii). In 2004 wild caught (n=5) and farmed (n=26) southern bluefin tuna (SBT) were collected. The mean concentration of total mercury on a fresh weight (fw) basis was 0.34 mg/kg in wild SBT and 0.31 mg/kg in farmed SBT. Concentrations of dioxins (PCDD/PCDFs) and polychlorinated biphenyls (PCBs) found in wild SBT were extremely low. The mean concentration (fw) of the sum of PCDD/PCDFs and the dioxin-like PCBs was 0.27 pg TEQ/g in wild SBT and 0.61 pg TEQ/g in farmed SBT. Mean PCB total concentration (fw basis) was 0.47 ng/g in wild SBT and 6.60 ng/g in farmed SBT. No detectable residues of any pesticide or antimicrobial compounds were found. All residues detected were within regulatory limits set by the Japanese Competent Authority, the Ministry of Health, Labour and Welfare (MHLW) and the Australian Competent Authority, Food Standards Australia New Zealand (FSANZ). The need for harmonisation of international product integrity procedures for bluefin tunas in international trade is highlighted. PMID- 18395243 TI - Quantitative structure-toxicity relationship study of lethal concentration to tadpole (Bufo vulgaris formosus) for organophosphorous pesticides. AB - In the present study more than 1,000 structural parameters of 41 organophosphorus pesticides (OPs) were calculated using the software ChemOffice 8.03 and Dragon 2.1. Then, with multivariate linear regression and best subset regression analyses, different equations were derived to calculate the lethal toxicity, LC(50), for these 41 organophosphorous pesticides found in tadpoles (Bufo vulgaris formosus). An equation was developed for all selected OPs, especially those with relatively low toxicity levels (LC(50)>4.5mM) that accounted for 89.09% of the variability in the toxic effect. The equation indicated that the main contributions to OPs toxicity with tadpoles were the electrostatic contribution qH(+) (maximum net positive H atomic charge), spatial autocorrelation (MATS7 m) and hydrophobicity (lgK(ow)), with the two former being the most important parameters. For OPs with high toxicity, however, different structural parameters were introduced. The following equation was developed with LC(50)<4.5mM. These equations implied that with different levels of toxicity there could have different mechanisms in the tadpole. Furthermore, the results showed that molecular structural parameters had a particular value in modeling chemical reactivity within a homologous series of compounds. PMID- 18395244 TI - Variation in particulate PAHs levels and their relation with the transboundary movement of the air masses. AB - The levels of particulate polycyclic aromatic hydrocarbons (PAHs) were determined with a fast analytical approach to study their seasonal variations at Menen (Belgium) during 2003; they were found to be 5-7 times higher in January, February and December, in comparison to May, June and August. The annual average concentration of the sum of 16 US Environmental Protection Agency (EPA) criteria PAHs was 6.7 ng/m3 and around 63% of it was found to be probably carcinogenic to humans. The application of diagnostic ratio and principal component analysis showed vehicular emission as a major source. An increased ratio of 'combustion PAHs' to 'total EPA-PAHs' during the winter season indicated towards combustion activities. Further, the differences in PAHs concentration were assessed with relation to backward air mass trajectories, which show that the levels of PAHs increase when there is an air mass movement from Central and Western Europe and a fall when the trajectories spend most of their 4-day time over the Atlantic Ocean or in the Arctic region. PMID- 18395245 TI - Effects of pyrite sludge pollution on soil enzyme activities: ecological dose response model. AB - A laboratory study was conducted to evaluate the response of soil enzyme activities (acid and alkaline phosphatase, beta-glucosidase, arylsulfatase, urease and dehydrogenase) to different levels of trace elements pollution in soils representative of the area affected by the pyrite sludge mining spill of Aznalcollar (Guadiamar basin, SW Spain). Three uncontaminated soils from the study area were mixed with different loads of pyrite sludge to resemble field conditions and criteria applied for reclamation practices following the pollution incident: 0% ("reference" or background level), 1.3% ("attention level", further monitoring required), 4% ("intervention level", further cleaning and liming required) and 13% (ten times the "attention level"). Enzyme activities were analysed 4, 7, 14, 21, 34 and 92 days after pollutant addition and those measured after 92 days were used to calculate the ecological dose value (ED50). Soil enzyme activities and pH decreased after the pyrite sludge addition with respect to the "reference level" (0% pyrite sludge), whereas soil bioavailable (DTPA extractable) trace elements concentration increased. Arylsulfatase, beta glucosidase and phosphatase activities were reduced by more than 50% at 1.3% pyrite sludge dose. Arylsulfasate was the most sensitive soil enzyme (in average, ED50=0.99), whereas urease activity showed the lowest inhibition (in average, ED50=7.87) after pyrite sludge addition. Our results showed that the ecological dose concept, applied to enzyme activities, was satisfactory to quantify the effect of a multi-metalic pollutant (pyrite sludge) on soil functionality, and would provide manageable data to establish permissible limits of trace elements in polluted soils. Additionally, we evaluate the recovery of enzyme activities after addition of sugar-beet lime (calcium carbonate) to each experimentally polluted soil. The amount of lime added to each soil was enough to raise the pH to the original value (equal to control soil), resembling field remediation practices. After lime amendment, soil recovery was still incomplete in terms of bioavailable trace elements. However, the recovery of soil enzyme activities varied widely, ranging from 0 to 100% depending on soil type and the specific enzyme. PMID- 18395247 TI - Axial stretch enhances sarcoplasmic reticulum Ca2+ leak and cellular Ca2+ reuptake in guinea pig ventricular myocytes: experiments and models. AB - Cardiac cellular calcium (Ca2+) handling is the well-investigated mediator of excitation-contraction coupling, the process that translates cardiac electrical activation into mechanical events. The reverse--effects of mechanical stimulation on cardiomyocyte Ca2+ handling--are much less well understood, in particular during the inter-beat period, called 'diastole'. We have investigated the effects of diastolic length changes, applied axially using a pair of carbon fibres attached to opposite ends of Guinea pig isolated ventricular myocytes, on the availability of Ca2+ in the main cellular stores (the sarcoplasmic reticulum; SR), by studying the rest-decay of SR Ca2+ content [Ca2+]SR, and the reloading of the SR after prior depletion of Ca2+ from the cell. Cells were loaded with Fura-2 AM (an indicator of the cytosolic 'free' Ca2+ concentration, [Ca2+]i), and pre conditioned by field-stimulation (2 Hz) at 37 degrees C, while [Ca2+]i transients and sarcomere length (SL) were recorded simultaneously. After reaching a steady state in the behaviour of observed parameters, stimulation was interrupted for between 5 and 60s, while cells were either held at resting length, or stretched (controlled to cause a 10% increase in SL, to aid inter-individual comparison). Thereafter, each cell was returned to its original resting length, followed by swift administration of 10mM of caffeine (in Na+/Ca2+-free solution), which causes the release of Ca2+ from the SR (caffeine), but largely prevents extrusion of Ca2+ from the cytosol to the cell exterior (Na+/Ca2+-free solution). By comparing the [Ca2+]i in cells exposed/not exposed to diastolic stretch of different duration, we assessed the rest-decay dynamics of [Ca2+]SR. To assess SR reloading after initial Ca2+ depletion, the same stretch protocol was implemented after prior emptying of the cell by application of 10mM of caffeine in normal Tyrode solution (which causes Ca2+ to be released from the SR and extruded from the cell via the Na+/Ca2+ exchanger; NCX). Axial stretch enhanced the rate of both rest-decay and reloading of [Ca2+]SR. Application of 40 microM streptomycin, a blocker of stretch-activated ion channels, did not affect the stretch-induced increase in SR reloading. This behaviour was reproduced in a computer simulation study, using a modified version of the 2006 Iribe-Kohl-Noble model of single cardiac myocyte Ca2+ handling, suggesting that stretch increases both Ca2+ leak from the SR and Ca2+ influx via the sarcolemma. This may have important implications for the mobilisation of Ca2+ in stretched cells, and could contribute to the regional 'matching' of individual cardiomyocyte contractility to dynamic, and regionally varying, changes in mechanical loads, such as diastolic pre-load, of cardiac tissue. PMID- 18395248 TI - Overexpression of HIPK2 circumvents the blockade of apoptosis in chemoresistant ovarian cancer cells. AB - OBJECTIVE: Chemoresistance, due to inhibition of apoptotic response, is the major reason for the failure of anticancer therapies. HIPK2 regulates p53-apoptotic function via serine-46 (Ser46) phosphorylation and activation of p53 is a key determinant in ovarian cancer cell death. In this study we determined whether HIPK2 overexpression restored apoptotic response in chemoresistant cancer cells. METHODS: Using cisplatin chemosensitive (2008) and chemoresistant (2008C13) ovarian cancer cell lines we compared drug-induced activation of the HIPK2/p53Ser46 apoptotic pathway. The levels of HIPK2, Ser46 phosphorylation, and PARP cleavage were detected by Western blotting. The p53Ser46 apoptotic commitment was evaluated by luciferase assay using the Ser46 specific AIP1 target gene promoter. The apoptotic pathway was detected by caspase-3, -8, and -9 activities. RESULTS: HIPK2 was expressed differently in sensitive versus chemoresistant cells in response to different chemotherapeutic drugs (i.e., cisplatin and adriamycin), though the p53Ser46 apoptotic pathway was not defective in chemoresistant 2008C13 cells. Thus, 2008C13 cells were resistant to cisplatin but sensitive to adriamycin-induced apoptosis through activation of the HIPK2/p53Ser46 pathway. HIPK2 knock-down inhibited the adriamycin-induced apoptosis in 2008C13 cells. Exogenous HIPK2 triggered apoptosis in chemoresistant cells, associated with induction of p53Ser46-target gene AIP1. CONCLUSIONS: HIPK2 is an important regulator of p53 activity in response to a chemotherapeutic drug. These results suggest that different drug-activated pathways may regulate HIPK2 and that HIPK2/p53Ser46 deregulation is involved in chemoresistance. Exogenous HIPK2 might represent a novel therapeutic approach to circumvent inhibition of apoptosis in treatment of chemoresistant ovarian cancers with wtp53. PMID- 18395246 TI - Cellular mechanisms of arrhythmogenic cardiac alternans. AB - Despite the strong association between mechanical dysfunction of the heart and sudden death due to arrhythmias, the causal relationship is not well understood. Cardiac alternans has been linked to arrhythmogenesis and can be mediated by intracellular calcium handling. Given the integral role intracellular calcium plays in contractile function, calcium-mediated alternans may represent an important mechanistic link between mechanical dysfunction and electrical instability. This relationship, however, is not well understood due to complex feedback between membrane currents, intracellular calcium, and contraction. This manuscript describes the cellular mechanisms of cardiac alternans. Through several pathways, calcium transient alternans is coupled to repolarization alternans that can form a substrate for reentrant excitation. Abnormal intracellular calcium cycling, either impaired release or impaired reuptake of sarcoplasmic reticulum calcium, is a cellular mechanism of calcium transient alternans. Thus, cardiac alternans is an important mechanistic link between mechanical dysfunction and sudden cardiac death. PMID- 18395249 TI - The role of cerium redox state in the SOD mimetic activity of nanoceria. AB - Cerium oxide nanoparticles (nanoceria) have recently been shown to protect cells against oxidative stress in both cell culture and animal models. Nanoceria has been shown to exhibit superoxide dismutase (SOD) activity using a ferricytochrome C assay, and this mimetic activity that has been postulated to be responsible for cellular protection by nanoceria. The nature of nanoceria's antioxidant properties, specifically what physical characteristics make nanoceria effective at scavenging superoxide anion, is poorly understood. In this study electron paramagnetic resonance (EPR) analysis confirms the reactivity of nanoceria as an SOD mimetic. X-ray photoelectron spectroscopy (XPS) and UV-visible analyses of nanoceria treated with hydrogen peroxide demonstrate that a decrease in the Ce 3(+)/4(+) ratio correlates directly with a loss of SOD mimetic activity. These results strongly suggest that the surface oxidation state of nanoceria plays an integral role in the SOD mimetic activity of nanoceria and that ability of nanoceria to scavenge superoxide is directly related to cerium(III) concentrations at the surface of the particle. PMID- 18395250 TI - Rapid effects of 17beta-estradiol on epithelial TRPV6 Ca2+ channel in human T84 colonic cells. AB - The control of calcium homeostasis is essential for cell survival and is of crucial importance for several physiological functions. The discovery of the epithelial calcium channel Transient Receptor Potential Vaniloid (TRPV6) in intestine has uncovered important Ca(2+) absorptive pathways involved in the regulation of whole body Ca(2+) homeostasis. The role of steroid hormone 17beta estradiol (E(2)), in [Ca(2+)](i) regulation involving TRPV6 has been only limited at the protein expression levels in over-expressing heterologous systems. In the present study, using a combination of calcium-imaging, whole-cell patch-clamp techniques and siRNA technology to specifically knockdown TRPV6 protein expression, we were able to (i) show that TRPV6 is natively, rather than exogenously, expressed at mRNA and protein levels in human T84 colonic cells, (ii) characterize functional TRPV6 channels and (iii) demonstrate, for the first time, the rapid effects of E(2) in [Ca(2+)](i) regulation involving directly TRPV6 channels in T84 cells. Treatment with E(2) rapidly (<5 min) enhanced [Ca(2+)](i) and this increase was partially but significantly prevented when cells were pre-treated with ruthenium red and completely abolished in cells treated with siRNA specifically targeting TRPV6 protein expression. These results indicate that when cells are stimulated by E(2), Ca(2+) enters the cell through TRPV6 channels. TRPV6 channels in T84 cells contribute to the Ca(2+) entry/signalling pathway that is sensitive to 17beta-estradiol. PMID- 18395253 TI - Circumvention of glucocorticoid resistance in childhood leukemia. AB - In this study, we determined if in vitro resistance to prednisolone and dexamethasone could be circumvented by cortivazol or methylprednisolone, or reversed by meta-iodobenzylguanidine in pediatric lymphoblastic and myeloid leukemia. As there were strong correlations between the LC50 values (drug concentration inducing 50% leukemic cell kill, LCK) of the different glucocorticoids and median prednisolone/methylprednisolone, prednisolone/dexamethasone and prednisolone/cortivazol LC50 ratios did not differ between the leukemia subtypes, we conclude that none of the glucocorticoids had preferential anti-leukemic activity. Meta-iodobenzylguanidine however, partially reversed glucocorticoid resistance in 19% of the lymphoblastic leukemia samples. PMID- 18395252 TI - TARC and IL-5 expression correlates with tissue eosinophilia in peripheral T-cell lymphomas. AB - The current study attempts to characterize the eosinophilia associated with T cell lymphomas and to investigate its possible relationship with the secretion of eosinophil-stimulating factors by lymphoma cells and/or intra-tumoral surrounding cells. Paraffin-embedded specimens from 50 patients diagnosed with peripheral T cell lymphomas, either unspecified (PTCL-U, n=30) or angioimmunoblastic (AITL, n=20) were morphologically assessed for intra-tumoral eosinophilia and analyzed by immunohistochemistry using specific antibodies directed against TARC, IL-5, RANTES, and eotaxin. The AITL and PTCL-U cases contained a mean of 147+/-41 and 102+/-37 eosinophils per 10 high power fields, respectively. Thirty-two of 47 cases (68%) showed IL-5-positive lymphoma cells while 15/50 (30%) tumors showed variable staining for TARC in scattered non-lymphoid cells with dendritic morphology. TARC and IL-5-positive cases possessed significantly more eosinophils. Our data indicate that IL-5 and TARC expression highly correlate with eosinophilia in T-cell lymphomas, suggesting that these chemokines are involved in the recruitment of eosinophils into the tumors. PMID- 18395254 TI - The BAFF-Interacting receptors of chickens. AB - The TNF superfamily cytokine BAFF has crucial roles in homoeostatic regulation of B cell populations in mammals. Similar effects on peripheral B cells have been reported for chicken as for mammalian BAFF. Unlike mammalian BAFF, chicken BAFF is produced by B cells, implying an autocrine loop and consequent differences in regulation of B cell homoeostasis. Understanding of these mechanisms requires investigation of BAFF-binding receptors in chickens. We identified and characterised chicken receptors BAFFR and TACI, but found that the gene encoding the third BAFF-binding receptor, BCMA, was disrupted, implying differences in mechanisms for maintenance of long-lived antibody responses. A BAFFR-Ig fusion protein expressed in vivo lowered B cell numbers, showing that it was functional under physiological conditions. We found changes in the ratio of BAFFR and TACI mRNAs in the bursa after hatch that may account for the altered requirements for B cell survival at this stage of development. PMID- 18395251 TI - Mitochondrial biogenesis and turnover. AB - Mitochondrial biogenesis is a complex process involving the coordinated expression of mitochondrial and nuclear genes, the import of the products of the latter into the organelle and turnover. The mechanisms associated with these events have been intensively studied in the last 20 years and our understanding of their details is much improved. Mitochondrial biogenesis requires the participation of calcium signaling that activates a series of calcium-dependent protein kinases that in turn activate transcription factors and coactivators such as PGC-1alpha that regulates the expression of genes coding for mitochondrial components. In addition, mitochondrial biogenesis involves the balance of mitochondrial fission-fusion. Mitochondrial malfunction or defects in any of the many pathways involved in mitochondrial biogenesis can lead to degenerative diseases and possibly play an important part in aging. PMID- 18395256 TI - The effects of Panax ginseng and Spirulina platensis on hepatotoxicity induced by cadmium in rats. AB - Cadmium is an environmental and industrial cumulative pollutant that affects many organs, specially the liver. The protective effect of Spirulina platensis and Panax ginseng on cadmium-induced oxidative stress and hepatotoxicity was evaluated in adult female Wistar albino rats. At the end of the 1-month experimental period, all animals were fasted for 12h and liver samples were taken for the determination of malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and nitric oxide (NO) levels. S. platensis and P. ginseng treatments showed marked decrease lipid peroxidation and increase of the endogenous antioxidants levels. The cadmium-induced histopathological changes were also minimized with the tested extracts. These results suggest that S. platensis and P. ginseng might play a role in reducing the toxic effect of cadmium and its antioxidant properties seem to mediate such a protective effect. PMID- 18395255 TI - Oligodeoxynucleotides containing CpG motifs (CpG-ODN) predominantly induce Th1 type immune response in neonatal chicks. AB - Earlier, we demonstrated that intramuscular administration of oligodeoxynucleotides containing CpG motifs (CpG-ODN) induces protection in neonatal chicks against a lethal challenge of Escherichia coli. However, the mechanism of induction of the protection was not clear. In an attempt to elucidate the mechanism of induced protection, we determined the kinetics of expression of cytokines/chemokines in the spleen and bursa of Fabricius of newly hatched chicks that had received intramuscular administration of CpG-ODN or non CpG ODN compared to saline-treated controls. SyBr green, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis of the RNA demonstrated increased expression of IL-1beta, IL-6, IL-8, IL-10, IL-18, IFN gamma and MIP-3alpha mRNAs in the spleen and; IL-10 and IFN-alpha in bursa of Fabricious of chicks that had received CpG-ODN. However, non-CpG ODN failed to induce any of the cytokine. The increased level of IL-18 and IFN-gamma but not IL 4 mRNA suggests that the administration of CpG-ODN elicits a Th1 biased immune response, which may be important in inducing protection against infections in neonatal chicks. To our knowledge, this is the first report evaluating the induction of cytokines/chemokines in neonatal chicks following administration of CpG-ODN. PMID- 18395257 TI - Acute and chronic effects of testosterone and 4-hydroxyandrostenedione to the crustacean Daphnia magna. AB - Steroid compounds have been globally detected in surface waters. The ecological impacts of these biologically active chemicals are largely unknown. Toxicity of testosterone and 4-hydroxyandrostenedione was assessed for the freshwater cladoceran Daphnia magna. Acute toxicity tests showed that 6.20 mg L(-1) of testosterone, the highest concentration tested, did not have effect on the daphnids, whereas 4-hydroxyandrostenedione had an EC(50) of 4.26 mg L(-1). Chronic toxicity tests were carried out using survival, body length, fecundity, and fertility as endpoints. Long-term testosterone exposure reduced D. magna fecundity and fertility at concentrations ranging from 0.31 to 2.48 mg L(-1). The significant decrease in fecundity was associated with an increase in aborted eggs. Long-term 4-hydroxyandrostenedione exposure at 0.84 mg L(-1) increased the mortality of the neonates. The chronic toxicity effects were observed at concentrations higher than the measured environmental concentrations of these compounds. Nevertheless, the reproductive impairment of the daphnids is likely to occur at environmental levels as an ultimate response to long-term exposure. PMID- 18395258 TI - Effects of nonylphenol on cholinesterase and carboxylesterase activities in male guppies (Poecilia reticulata). AB - Compared to the estrogenic effects of 4-nonylphenol (NP), there is little data available on other potential toxic effects of NP in aquatic animals. The effects of NP on cholinesterase (ChE) and carboxylesterase (CbE) activities of male guppies exposed to 10, 60, 150, or 300 MUg L(-1) NP were examined after 1, 2, 4, and 7 days of treatment. A significant muscle ChE inhibition, that used acetylthiocholine iodide as a substrate, was noted in male guppies in all NP treatment groups after a 4-day exposure, and 60 and 150 MUg L(-1) of NP treatment groups after a 7-day exposure. All guppies exposed to 300 MUg L(-1) NP died during the 7-day treatment. However, there was no significant inhibition of muscle ChE that used butyrylthiocholine iodide as a substrate in male guppies for any NP treatments in different exposure times. There were no CbE activity differences in livers of male guppies among NP treatment groups after different exposure times. This is the first report showing the ChE activity inhibition by NP in fish. Further mechanistic studies are needed to define how NP directly or indirectly alters ChE activities at molecular level. The implication of ChE inhibition of NP on potential impacts of aquatic animals also warrants further research. PMID- 18395259 TI - Cytogenetic alterations induced by SPL (spent potliners) in meristematic cells of plant bioassays. AB - Spent potliners (SPL) is solid waste generated by the aluminum industry during the manufacture of aluminum metal. Information on the cytotoxicity effect of SPL is necessary to facilitate understanding of their action on organisms and to subsidize environmentally correct solutions. Thus, the aim of the present investigation is to compare the effect of SPL on meristematic cells of Allium cepa and Zea mays and also to discuss the mechanisms of SPL cytotoxicity involved. A strong inhibition on root growth in higher SPL concentrations has been observed in both A. cepa and Z. mays. For cytogenetic analysis, the results showed a reduction of mitotic index and increase of different abnormalities as the SPL concentration increased. We observed bridges, chromosome fragments, stickiness, multipolar anaphase, later segregation and cell death. In general, it was possible to observe an increase of different abnormalities as the SPL concentration increased. It is obvious from the results of the present investigation that SPL is cytotoxic on meristematic cells of plant tests (A. cepa and Z. mays). PMID- 18395260 TI - Searching for autoimmunity in "antibiotic-refractory" Lyme arthritis. AB - In two recent articles published in Molecular Immunology, Steere and colleagues continue their search for an autoimmune mechanism of arthritis in patients who have failed short-course antibiotic therapy for Borrelia burgdorferi, the spirochetal agent of Lyme disease. As in previous attempts, the authors fail to elucidate a molecular mimicry mechanism for the putative autoimmune process, leading to the conclusion that there is no credible scientific evidence for a post-infectious autoimmune mechanism of arthritis in chronic Lyme disease. PMID- 18395261 TI - Reshuffling of ancient peptide binding motifs between HLA-DRB multigene family members: old wine served in new skins. AB - In most primate species, the class II region of the Major Histocompatibility Complex (MHC) displays diversity with regard to gene copy number and combination of DRB genes present per region configuration. Some of these loci exhibit extremely high levels of allelic variability, whereas others display only moderate levels of polymorphism. To understand the evolutionary history of the various HLA-DR region genes, a large number of full-length sequences of rhesus macaques, chimpanzees and humans were determined. The exon-intron organisation of the DRA gene, displaying only low levels of polymorphism, appears to have been highly conserved during primate evolution. The physical length of various DRB genes/alleles, however, fluctuates significantly in primates due to the presence of indels (insertions/deletions), mainly mapping to intron 1. Phylogenetic evidence supports the notion that the generation of new DRB genes is a dynamic and steadily ongoing process. Indeed, most of the primate DRB alleles investigated represent relatively young entities, possessing species-unique sequences. This seems to contradict the current view that the highly similar peptide binding motifs of many HLA-, Patr- and Mamu-DR molecules, encoded by exon 2 of the DRB gene, represent old entities, which predate primate speciation. As no evidence was found for convergent evolution, the combination of these two observations indicates that ancient peptide binding motifs are frequently reshuffled among duplicated members of the HLA-DRB multigene family. PMID- 18395262 TI - Identification of outer membrane protein ompR from rickettsia-like organism and induction of immune response in Crassostrea ariakensis. AB - Rickettsia-like organism (RLO) caused mass mortality of oysters, but little is known about the protective immune response to this microorganism. The present study was undertaken to identify a gene, ompR, encoding an outer membrane protein of rickettsia-like organism from oyster Crassostrea ariakensis. The role of this protein in promoting immune responses was characterized through analyzing the interaction between RLO and oyster. The results indicated: (i) full-length DNA of ompR is 531 bp and encodes 176 amino acid residues. Theoretical isoelectric point and molecular weight for the ompR protein are 9.76 and 19.76 kDa, respectively; (ii) the recombinant ompR was successfully expressed in Escherichia coli BL21 (DE3) cells, and the titre of anti-ompR antibody raised against rabbits was about 1:4100. A specific immunoreactive band was detected when anti-ompR antibody was opposed to the total outer membrane proteins of RLO; (iii) the expression level of TNF-alpha (tumor necrosis factor-alpha) and Myd88 (myeloid differentiation factor 88) in hemocytes was induced by ompR, whereas TGF-beta (transforming growth factor-beta) was not; (iv) in hemocytes monolayers, a rapid and persistent increase in the level of phosphorylated P38 and a large decrease in the level of phosphorylated JNK were induced by ompR, whereas the level of phosphorylated ERK did not change with ompR incubation; (v) the DNA binding activity of NF-kappaB (nuclear factor kappaB) in hemocytes increased after ompR stimulation. PMID- 18395263 TI - Cerebrospinal fluid levels of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in subacute sclerosing panencephalitis. AB - OBJECTIVES: To investigate the brain inflammation and damage in subacute sclerosing panencephalitis (SSPE), the cerebrospinal fluid (CSF) concentrations of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were determined in SSPE patients. METHODS: CSF MMP-9 and TIMP-1 levels were measured in 23 patients with SSPE in Papua New Guinea by ELISA. RESULTS: CSF MMP-9 levels and MMP-9/TIMP-1 ratios of SSPE patients were significantly higher than controls (p<0.001 and p=0.005, respectively). There were no significant differences in CSF TIMP-1 levels between SSPE patients and controls. CONCLUSIONS: Previous studies suggested that CSF MMP-9 levels reflect inflammatory damage to the brain. Our findings suggest that the MMP-9 level in CSF is an indicator of inflammatory damage to the brain in SSPE. PMID- 18395264 TI - Comparison of the sensitivity and specificity of two whole blood interferon-gamma assays for M. tuberculosis infection. AB - OBJECTIVES: To compare the sensitivity and the specificity of the QuantiFERON-TB Gold (QFT-G) and QuantiFERON-TB Gold In Tube (QFT-GIT) diagnostic tests for Mycobacterium tuberculosis infection. METHODS: One-hundred patients with culture and/or PCR confirmed M. tuberculosis infection and 168 volunteers with no risk factors for M. tuberculosis infection were tested to estimate sensitivity and specificity, respectively. RESULTS: Analysis of data from the tuberculosis (TB) patients with valid results found the sensitivity of QFT-GIT (92.6%, 87/94) to be significantly higher than that for the QFT-G test (81.4%, 79/97; p=0.023). The specificity of both QFT-GIT and QFT-G was 98.8% (CI: 95.1%-99.8%) with 2 of the 160 low risk subjects with valid results for both tests being positive. Data analysis confirmed the manufacturer's recommended test cut-off as being optimal, but identified higher sensitivity could be obtained by using a lower cut-off, with only a moderate decrease in specificity. CONCLUSIONS: The QFT-GIT test had enhanced sensitivity for detection of M. tuberculosis infection over the QFT-G test, whilst maintaining equivalent high specificity. The logistic benefits of the QFT-GIT test format, as well as its higher sensitivity, should enable enhanced TB control. PMID- 18395265 TI - The alternating current polarographic behavior and determination of lansoprazole and omeprazole in dosage forms and biological fluids. AB - The alternating current (ACt) polarographic behavior of lansoprazole (LNS) and omeprazole (OMP) was studied in Britton Robinson buffers (BRb) over the pH range 4.1-11.5. In BRb of pH 9.6 and 10.5, well-defined ACt peaks were obtained for both LNS and OMP, respectively. The current-concentration plots were rectilinear over the ranges of 0.4-20 microg mL(-1) and 0.2-10 microg mL(-1) for LNS and OMP respectively. The minimum detection limits (S/N=2) were 0.02 microg mL(-1) (5.4x10(-8) M) and 0.01 microg mL(-1) (2.9x10(-8) M) for LNS and OMP, respectively. The proposed method was successfully applied to the analysis of the two drugs in their commercial capsules. The average percent recoveries were favorably compared to those obtained by reference methods. Co-administered drugs such as naproxen and methotrexate did not interfere with the proposed method. The proposed method was further extended to the in-vitro determination of the lansoprazole in spiked plasma, the percentage recoveries was 98.47+/-1.29 (n=4). The pathway for the electrode reaction for both drugs involved reduction of the sulphonyl group into the corresponding thiol group at the Dropping Mercury Electrode. The advantages of the method were time saving and more sensitive than the other published voltammetric method. Yet The present study is the first report on the use of alterating current polarography (ACt) in this respect. PMID- 18395266 TI - A digital sequence method of dynamic olfactory characterization. AB - Measurements of system dynamics, such as input-output frequency response estimation, have been widely used in neuroscience. Dynamic characterization of sensory systems has been particularly useful because both the amplitude and time dependent properties of sensory input signals can often be accurately controlled. However, chemoreceptors have proved less amenable to these approaches because it is often difficult to accurately modulate or measure chemical concentration at a sensory receptor. New methods of dynamic olfactory stimulation have recently been introduced that combine controlled mechanical release of odorant with detection by photoionization of surrogate tracer gas mixed with the odorant. We have developed a modified version of this approach based on rapid binary switching of odorant flow using pseudo-random binary signals (maximum-length sequences, or M sequences) generated by a software shift register. This system offers several advantages over previous methods, including higher frequency range stimulation, experimental simplicity and the possibility of computational efficiencies. We show that there is predictable dynamic odorant concentration at the sensory receptor and we explore the stimulation parameters as functions of total air flow rate and spatial location. A typical application of the system is shown by measuring the frequency response function of a Drosophila electroantennogram. PMID- 18395267 TI - Specific depressive symptoms and disorders as associates and predictors of suicidal acts across adolescence. AB - OBJECTIVE: To examine the role of depressive symptoms and disorders as associates and predictors of suicidal acts across adolescence. METHOD: A representative sample of Norwegian school students (N = 2464, mean age 13.7 years) in grades 8 and 9 was reassessed after one year (T2) with the same questionnaire. All high scorers of depressive symptoms on the Mood and Feelings Questionnaire (MFQ) at T2 were defined as cases. One control from low or middle scorers, matched for age and gender, was randomly assigned to every two cases. This subset (n = 345) was diagnostically assessed by face-to-face K-SADS-PL interviews (mean age = 14.9 years). The same subset was reassessed after 5 years (T3) by using the same questionnaire (n = 252, mean age = 20.0 years) and telephone K-SADS-PL interviews (n = 242). The participation rate was 76.9% (n = 265). RESULTS: Cognitive symptoms dominated the depressive symptom profile among suicide attempters, irrespective of age and time. Among younger adolescents, suicidal thoughts and acts of self-harm without suicidal intent were associated with suicidal acts. Recurrent thoughts about death, hopelessness, disturbed concentration and middle insomnia were associates of suicidal acts among older adolescents. Worthlessness by 15 years was a significant predictor of suicidal acts between 15 to 20 years. MDD and a depressive episode, not otherwise specified, continued to be significant associates among younger adolescents, while dysthymia by 15 years remained a predictor of suicidal acts between 15 to 20 years, even when controlled for depressive symptoms. CONCLUSIONS: Self-harm without suicidal intent, middle insomnia, cognitive depressive symptoms and a formal psychiatric diagnosis of any depressive disorder should alert professionals in the risk assessment of suicidal adolescents. PMID- 18395268 TI - Bioelectric impedance is a better indicator of obesity in men with schizophrenia than body mass index. AB - Body mass index (BMI) is commonly used as an indicator of obesity, although in both clinical and research settings the use of bioelectric impedance analysis (BIA) is commonplace. The purpose of this study was to examine the relationship between BMI, BIA and percentage body fat to determine whether either is a superior indicator of obesity in men with schizophrenia. The reference method of deuterium dilution was used to measure total body water and, subsequently, percentage body fat in 31 men with schizophrenia. Comparisons with the classification of body fat using BMI and BIA were made. The correlation between percentage body fat and BMI was 0.64 whereas the correlation between percentage body fat and BIA was 0.90. The sensitivity and specificity in distinguishing between obese and overweight participants was 0.55 and 0.80 for BMI and 0.86 and 0.75 for BIA. BIA proved to be a better indicator of obesity than BMI. BMI misclassified a large proportion of men with schizophrenia as overweight when they had excess adiposity of sufficient magnitude to be considered as obese. Because of the widespread use of BMI as an indicator of obesity among people with schizophrenia, the level of obesity among men with schizophrenia may be in excess of that previously indicated. PMID- 18395269 TI - Anti-inflammatory action of alpha-melanocyte stimulating hormone (alpha-MSH) in anti-CD3/CD28-mediated spleen and CD4(+)CD25(-) T cells and a partial participation of IL-10. AB - alpha-Melanocyte stimulating hormone (alpha-MSH) has been shown to inhibit the production and the effects of pro-inflammatory cytokine by inflammatory cells in innate immunity. We have determined whether alpha-MSH inhibits anti-CD3/CD28 mediated spleen cells and CD4(+)CD25(-) T cells proliferation and its mechanism of action. The proliferation of anti-CD3/CD28-mediated spleen cells and CD4(+)CD25(-) T cells markedly were suppressed by 50-100 nM and 5-100 nM alpha MSH, respectively. alpha-MSH (100 nM) increased the production of the anti inflammatory cytokine IL-10 and decreased the production of the pro-inflammatory cytokine IL-2 and IFN-gamma from CD4(+)CD25(-) T cells. Moreover, anti-IL-10 blocking Ab decreased the inhibitory effects of anti-CD3/CD28-mediated spleen cells and CD4(+)CD25(-) T cells proliferation by alpha-MSH, indicating a partial participation of IL-10 in its mechanism of inhibitory action. These results suggest that alpha-MSH may be useful for treatment of autoimmune diseases and transplantation involving innate and adaptive immunity. PMID- 18395270 TI - The universal newborn hearing screening program at the University Hospital of Ferrara: focus on costs and software solutions. AB - In the present paper, the authors report the results of the Universal Newborn Hearing Screening (UNHS) project at the University Hospital of Ferrara. A total of 6,759 full-term newborns and a total of 1,016 NICU babies were tested at the University Hospital of Ferrara, from January 2000 to December 2006. The paper presents information from clinically acceptable screening procedures developed and tested during the 6 years of the program and addresses two questions pertinent to hearing screening: (i) the cost-estimate of a UNHS program based on European economical and administration premises and (ii) the development of a database-structure for the evaluation of the UNHS/NHS performance and the individual patient tracking. PMID- 18395271 TI - Adenotonsillar pathology in post-transplant patients. AB - OBJECTIVE/HYPOTHESIS: The purpose of this study is to evaluate the efficacy of adenotonsillar hypertrophy, Epstein-Barr virus (EBV) titers, and flow cytometry in the evaluation of post-transplant lymphoproliferative disease (PTLD) in an at risk pediatric population and to propose flow cytometry is the most effective means of diagnosis and directed treatment. STUDY DESIGN/METHODS: The authors conducted a retrospective chart review of all pediatric transplant patients referred for adenotonsillectomy at a tertiary care center between May 2000 and May 2003. Thirteen patients were identified. All tonsils were evaluated for follicular lymphoid hyperplasia and atypical lymphoid cells. Seven of thirteen samples were stained for EBV RNA, and twelve of thirteen samples were tested using flow cytometry. Positive flow cytometry diagnosis was based on the presence of atypical lymphocytes, gene arrangement, or both. RESULTS: Seven of thirteen (7/13) examined patients were found to have adenotonsillar hypertrophy. Ten of thirteen (10/13) patients had elevated EBV serum titers. Four of thirteen (4/13) patients were found PTLD positive by flow cytometry. Of the four positive patients, all four patients were found to have elevated EBV titers and only two patients had adenotonsillar hypertrophy. CONCLUSIONS: Adenotonsillar hypertrophy is not a sensitive indicator of PTLD. EBV serum titers are sensitive but nonspecific. Flow cytometry is the most effective diagnostic modality and allows physicians to tailor treatment to specific forms of PTLD. PMID- 18395272 TI - The effect of implantation age on visual attention skills. AB - OBJECTIVE: The aim of this study is to evaluate the visual attention skills of deaf children with cochlear implants and to determine whether age at implantation had any impact on visual attention skills. SUBJECT AND METHODS: Thirty children with cochlear implants aged 6 and 11 years were evaluated and were compared with age matched 36 normal hearing children. The children with cochlear implants were analyzed in to two groups according to their age at implantation; under and at/over 4 years old. The Gordon Diagnostic System (GDS) was used in order to evaluate visual attention skills of cochlear implanted (Group 1 and Group 2) and children with normal hearing. The number of correct responses, extraneous button presses (commission errors) and the number of omission errors or the failure to respond to target stimuli which were recorded by GDS was used to evaluate the visual attention skills of the subjects. Also six different types of commission errors (19X, XX9, XX1, X1X, X9X, XXX) were analyzed and compared both for normal and cochlear implant user children. RESULTS: Concerning the total number of correct response, omission and commission errors no statistical difference was found between two implanted groups. Besides, both groups with cochlear implantation differed from normal hearing peers on the total number of correct responses, omission errors (p<0.017). According to these findings cochlear implanted children had less correct responses and had more omission errors compared with control group. These findings clearly show that children with cochlear implants performed poorly on visual attention task which requires constant attention together with the ability to exert impulse control despite they hear enhanced sensory information by cochlear implants. Concerning the types of commission errors Group 1 and 2 performed similarly in all types of errors (p>0.017). Besides, Group 1 and 2 made more errors in types 19X, XX9 and XXX, XX9 compared with the control group (p<0.017). Group 2 who had their implants after age of 4 years old had more commission errors than the control group (p<0.017) and positive correlation was found between the total commission errors and XX9 error type with age at implantation. These findings show that children who had their implants at older ages made more commission errors and XX9 type of commission error. The children who were implanted after 4 years old were less mature; unsuccessful in controlling their impulsive behaviors and more careless where they can not get enough benefit from social and environmental motives compared with their normal hearing peers. CONCLUSION: This present research points out the positive effect of early implantation on visual attention and the possible adversities such as impulsivity, over-impatience and attention deficits which have been previously suggested as preventive or slowing down factors for the pace of auditory-verbal therapy for cochlear implantees. Assuming that the results of this study support the theory of insufficiency which suggested that the deprivation in auditory system would also adversely affect cognitive structures involved in intermodal processing the importance of early implantation is underlined. If early cochlear implantation will be carried out especially at the very critical period of language development (before age of four), this will help the child, in his/her future life, not only with his/her hearing skills and communication but for his/her attention skills and behavior. PMID- 18395273 TI - Tonsillectomy versus tonsillotomy performed with scissors in children with tonsillar hypertrophy. AB - OBJECTIVE: We sought to determine the morbidity and the short and long-term effectiveness of tonsillotomy (partial tonsillectomy) performed with scissors compared with conventional cold knife (total) tonsillectomy. METHODS: We conducted a 2-year prospective non-randomized study at a tertiary children's hospital. One thousand and twenty-three children were recruited (243 in the tonsillotomy and 780 in the tonsillectomy group). Age, sex, weight, and time of operation, immediately post-operative complications and pain were recorded. Eighteen months after the operation data on weight, parents' satisfaction and recurrence of obstructed symptoms was obtained in 60 randomly selected children of each group. RESULTS: Post-operative complications rates were very low in both groups and their differences did not reach statistical significance. However, tonsillotomies were significantly better than tonsillectomies in relation to post operative pain the 1st (P<0.001) and the 7th (P<0.001) post-operative day, and lasted an average of 2.5 min less. Concerning long-term effects, both methods resulted in an increase of patients' weight without the difference in weight increase be statistically significant. In addition, parents' satisfaction as well as oral malodour do not seem to differ significantly. Finally, although twice as many children in the tonsillotomy group had a recurrence of snoring compared to tonsillectomy group, only in about half of them was the problem regarded significant enough by their parents to require medical consultation or reoperation. At the end, an estimated 3.5% of the tonsillotomy group was offered a tonsillectomy in the long term. CONCLUSION: This trial shows that scissors tonsillotomy in combination with an adenoidectomy is a viable therapeutic option with less immediate post-operative morbidity than tonsillectomy in young children whose sleep disorder breathing is due to a combination of tonsilar and adenoid hypertrophy. However, parents should be informed of the possibility of regrowth and reoperation. PMID- 18395274 TI - A role for the spine apparatus in LTP and spatial learning. AB - Long-term potentiation (LTP) of synaptic strength is a long-lasting form of synaptic plasticity that has been linked to information storage. Although the molecular and cellular events underlying LTP are not yet fully understood, it is generally accepted that changes in dendritic spine calcium levels as well as local protein synthesis play a central role. These two processes may be influenced by the presence of a spine apparatus, a distinct neuronal organelle found in a subpopulation of telencephalic spines. Mice lacking spine apparatuses (synaptopodin-deficient mice) show deficits in LTP and impaired spatial learning supporting the involvement of the spine apparatus in synaptic plasticity. In our review, we consider the possible roles of the spine apparatus in LTP1 (protein synthesis-independent), LTP2 (translation-dependent and transcription independent) and LTP3 (translation- and transcription-dependent) and discuss the effects of the spine apparatus on learning and memory. PMID- 18395275 TI - Effects of visual cortex lesions on perceptual grouping in rats. AB - Neural mechanisms mediating perceptual grouping serve to enhance associations among stimulus elements, thereby establishing unified forms. The goals of the present study were to identify cortical areas necessary to perceptually group spatially isolated elements, and to determine if these areas are distinct from regions necessary for the discrimination of simple, solid forms. Rats were trained to discriminate horizontal and vertical lines that were either solid or composed of disjunct elements in which discrimination required perceptual grouping by proximity. Psychophysical procedures established the limits at which proximity served as a cue for grouping. Following perceptual measurements, ablations were made to selective sites within visual cortex. Lesions within area 17 or area 18A, including their interface, produced nearly complete impairment of solid line discrimination as well as perceptual grouping at all levels of proximity, whereas lesions to areas 18 or the far lateral extent of area 18A no effect on these perceptual capacities. These results indicate that grouping by proximity requires early visual processing areas, and shares cortical areas necessary for simple pattern discrimination. These results suggest that mechanisms of grouping modify primary stimulus representations, constructing a pattern of activity functional similar to that elicited by solid forms. PMID- 18395276 TI - Effect of atrazine on metamorphosis and sexual differentiation in Xenopus laevis. AB - There is a growing international concern that commonly used environmental contaminants have the potential to disrupt the development and functioning of the reproductive system in amphibians. One such chemical of interests is the herbicide atrazine. Effects of atrazine on sex differentiation were studied using wild-type Xenopus laevis tadpoles and all-ZZ male cohorts of X. laevis tadpoles, produced by mating wild-type ZZ male to sex-reversed ZZ male (female phenotype). Stage 49 tadpoles were exposed to 0.1-100 ppb atrazine or 0.27 ppb (1 nM) 17beta estradiol (E(2)) until all larvae completed metamorphosis (stage 66). Metamorphosis, gonadal morphology and histology, CYP19 (P450 aromatase) mRNA induction, and hepatic vitellogenin (VTG) induction were investigated. Effects of atrazine on VTG-induction were also assessed in vitro in primary-cultured X. laevis hepatocytes. Atrazine had no effect on metamorphosis of developing wild type or all-male X. laevis larvae. Statistical increase in female ratios was observed in 10 and 100 ppb atrazine groups in comparison with control group. While no hermaphroditic froglet was observed in all atrazine groups. In ZZ males, sex reversal was induced by 0.27 ppb E(2), but not by atrazine at concentrations of 0.1 and 1.0 ppb. In addition, neither P450 aromatase mRNA in the gonad nor hepatic VTG were induced by atrazine. Furthermore, VTG was not induced by 1000 ppb atrazine in primary-cultured hepatocytes. Our results indicate that female ratios in developing X. laevis tadpoles were increased by 10 and 100 ppb atrazine under the present experimental conditions. While the other endpoints showed no effect in the range of 0.1-100 ppb atrazine. These results suggest that effect of atrazine on sexual differentiation was not caused by estrogenic action and has no induction ability of P450 aromatase gene in gonad. PMID- 18395277 TI - Market incentives and pharmaceutical innovation. AB - I study the impact of the Orphan Drug Act (ODA), which established tax incentives for rare disease drug development. I examine the flow of new clinical drug trials for a large set of rare diseases. Among more prevalent rare diseases, the ODA led to a significant and sustained increase in new trials. The impact for less prevalent rare diseases was limited to an increase in the stock of drugs. Tax credits can stimulate R & D; yet because they leave revenue margins unaffected, tax credits appear to have a more limited impact on private innovation in markets with smaller revenue potential. PMID- 18395278 TI - Ultrafiltration-based techniques for rapid and simultaneous concentration of multiple microbe classes from 100-L tap water samples. AB - This study focused on ultrafiltration as a technique for simultaneously concentrating and recovering viruses, bacteria and parasites in 100-L drinking water samples. A chemical dispersant, sodium polyphosphate, and Tween 80 were used to increase microbial recovery efficiencies. Secondary concentration was performed to reduce sample volumes to 3-5 mL for analysis using tissue culture, microscopy, and real-time PCR and RT-PCR. At seeding levels of 100-1000 (CFU, PFU, oocysts, or particles), a "high-flux" ultrafiltration procedure was found to achieve mean recoveries of 51-94% of simultaneously seeded MS2 bacteriophage, echovirus 1, Salmonella enterica subsp. enterica serovar Typhimurium, Bacillus atrophaeus subsp. globigii endospores, Cryptosporidium parvum oocysts, and 4.5 mum microspheres. When 4-7% of the final sample concentrate volume was assayed using real-time PCR and RT-PCR, overall method sensitivities were <100 C. parvum oocysts, <240 PFU echovirus 1, <100 CFU Salmonella and approximately 160 CFU B. atrophaeus spores in 100-L drinking water samples. The "high-flux" ultrafiltration procedure required approximately 2 h, including time required for backflushing. Secondary concentration procedures required an additional 1-3 h, while nucleic acid extraction and real-time PCR procedures required an additional 2-2.5 h. Thus, this study demonstrated that efficient recovery and sensitive detection of diverse microbes in 100-L drinking water samples could be achieved within 5-8 h using ultrafiltration, rapid secondary processing techniques, and real-time PCR. PMID- 18395279 TI - Development of a rapid and sensitive immunoassay for detection and subsequent recovery of Bacillus anthracis spores in environmental samples. AB - Bacillus anthracis is considered a major threat as an agent of bioterrorism. B. anthracis spores are readily dispersed as aerosols, are very persistent, and are resistant to normal disinfection treatments. Immunoassays have been developed to rapidly detect B. anthracis spores at high concentrations. However, detection of B. anthracis spores at lower concentrations is problematic due to the fact that closely related Bacillus species (e.g., B. thuringiensis) can cross-react with anti-B. anthracis antibodies, resulting in false positive detections. Subsequent polymerase chain reaction (PCR) analysis is required to differentiate virulent strains. We report here on a protocol for the rapid, sensitive detection of B. anthracis spore using the Integrating Waveguide Biosensor followed by a method for the rapid release and germination of immunocaptured spores. A detection limit of ca. 10(3) spores was achieved by incubating spores simultaneously with capture and detection antibodies ("liquid-phase" assay) prior to capture on capillary tubes/waveguides. Subsequent incubation with BHI broth directly in capillary tubes allowed for rapid germination, outgrowth, and release of spores, resulting in vegetative cells for PCR analysis. PMID- 18395280 TI - Comparative evaluation of radiochemotherapy with temozolomide versus standard-of care postoperative radiation alone in patients with WHO grade III astrocytic tumors. AB - Outcome after radiochemotherapy (RCHT) with temozolomide (TMZ) versus radiotherapy (RT) for WHO grade III astrocytic tumors was evaluated. No significant difference in overall survival or progression-free survival between both groups was calculated. RCHT seems not to result in an improved outcome. Further randomized studies are needed to support these results. PMID- 18395281 TI - A study of EEG coherence in DCD children during motor synchronization task. AB - This paper investigates the hypothesis that the coordination difficulties of DCD children are associated with an increased coherence in the cortical motor regions, which persists with age. Forty-eight children participated in the study (24 DCD and 24 Controls). Their ages ranged from 8 to 13 years, divided into three groups (8-9, 10-11, and 12-13 years old). Children were required to perform finger flexion or extension either in synchrony or in syncopation with a rhythmic metronome, while a 32-channel EEG was recorded. Along with stability measures of motor performance, we analyzed the spectral EEG coherence between intrahemispheric (left frontal/left central; left central/left parietal) and interhemispheric (left central/right central) sites. Spectral coherence assesses functional coupling between distant areas of the brain. Two frequency bands related to sensorimotor activation were chosen: alpha (8-12 Hz) and beta (12-30 Hz). The synchrony task was chosen as a rest condition against which the two syncopation conditions at 0.5 Hz and 1.3 Hz were contrasted. For intrahemispheric comparison, 8-9-year-old DCD children showed that coherence between fronto central regions increased for both rhythms and conditions, as compared to controls. No difference was found for interhemispheric comparisons. As frontal sites are related to motor planning, our results suggest that youngest DCD children were forced to maintain a high level of pre-programming to compensate for the difficulties caused by the perceptual-motor requirements of the task in light of their coordination disorder. PMID- 18395282 TI - Physical fitness and health indices in children, adolescents and adults with high or low motor competence. AB - The overall purpose of the study was to examine if individuals with low motor competence achieve age-adequate fitness and health. A group of 149 children, adolescents, and adults with low or high motor competence participated in motor, fitness, and health assessments. Individuals with low motor competence did not differ on their basic physiological health parameters, but they had less optimal levels of overall health and fitness indices than those with high motor competence. As a function of age, musculoskeletal fitness was significantly compromised for the low motor competence group. The metabolic indices suggested that the low motor competence group had significantly higher BMI's compared to the high motor competence group. Motor skills and static balance were significant predictors of the BMI. Exercise intensity differed between children in the low and high motor competence group. The findings suggest that individuals with low motor competence have compromised health-related fitness. In order to discriminate between individuals with high and low motor competence, fitness assessment should include at least back extension, curl ups, and sit and reach. In addition, health-related fitness measurements such as BMI, waist circumference, blood lipid profile and bone mineral density are also recommended. PMID- 18395283 TI - Functionalized micelles from block copolymer of polyphosphoester and poly(epsilon caprolactone) for receptor-mediated drug delivery. AB - Cellular specific micellar systems from functional amphiphilic block copolymers are attractive for targeted intracellular drug delivery. In this study, we developed reactive micelles based on diblock copolymer of poly(ethyl ethylene phosphate) and poly(epsilon-caprolactone). The micelles were further surface conjugated with galactosamine to target asialoglycoprotein receptor (ASGP-R) of HepG2 cells. The size of micellar nanoparticles was about 70nm in diameter, and nanoparticles were negatively charged in aqueous solution. Through recognition between galactose ligands with ASGP-R of HepG2 cells, cell surface binding and internalization of galactosamine-conjugated micelles were significantly promoted, which were demonstrated by flow cytometric analyses using rhodamine 123 fluorescent dye. Paclitaxel-loaded micelles with galactose ligands exhibited comparable activity to free paclitaxel in inhibiting HepG2 cell proliferation, in contrast to the poor inhibition activity of micelles without galactose ligands particularly at lower paclitaxel doses. In addition, population of HepG2 cells arrested in G2/M phase was in positive response to paclitaxel dose when cells were incubated with paclitaxel-loaded micelles with galactosamine conjugation, which was against the performance of micelles without galactose ligand, owing to the ligand-receptor interaction. The surface functionalized micellar system is promising for specific anticancer drug transportation and intracellular drug release. PMID- 18395284 TI - [The effectiveness of constraint-induced movement therapy for children with hemiplegia following acquired brain injury]. AB - Constraint-induced movement therapy is a promising technique for improving upper limb function in adults with hemiplegia. It involves restraint of the non involved limb and intensive movement practice with the paretic limb. Although the technique has been applied successfully to children with cerebral palsy, only two studies have used it in children with acquired brain injury. OBJECTIVE: To assess the feasibility and efficacy of constraint-induced movement therapy in children with acquired brain injury. METHODS: We used a single-subject experimental design in three children (aged five at the time of the intervention) with hemiplegia in the chronic phase following acquired brain injury. The intervention involved restraint of the unaffected arm with a Mayo Clinic elbow brace for seven hours a day in a hospital setting, together with three hours a day of physical and occupational therapy rehabilitation for five days a week for two weeks. The children were assessed twice at baseline and then once immediately post-treatment and again two months post-treatment. Assessment included a range of timed, quantitative measures of upper limb use, assessment of unilateral spatial neglect and qualitative assessment by therapists and parents in terms of activities of daily living. RESULTS: The three children completed the full protocol and improved significantly in all timed, quantitative tests of motor function. These improvements were partially maintained at two months. No improvement in unilateral spatial neglect was found in the paper-and-pencil tasks, although less spatial neglect was observed in activities of daily living such as eating and walking. CONCLUSION: Constraint-induced movement therapy appears to be both feasible and efficient in children with acquired brain injury. PMID- 18395285 TI - [What is the interest of early mobilization after total hip arthroplasty? Development of French guidelines for clinical practice]. AB - AIMS: To develop clinical practice guidelines for early mobilisation after total hip replacement (THR). METHOD: We used the French Society of Physical and Rehabilitation Medicine (Sofmer) methodology, which associates a systematic review of the literature, the collection of information regarding current clinical practice and external review by a multidisciplinary expert panel. RESULTS: Recommending early mobilisation after THR is not established by a review of the literature. A survey of French clinical practice allows for recommending early mobilisation in the context of complex hip issues. Trials with good methodology must be developed to evaluate the interest of early functional mobilisation corresponding to when patients first stand and take their first steps after surgery. These trials should focus mainly on the final pain, functional status, and reduction of handicap. PMID- 18395286 TI - [Comparative study of dyspraxia: neuropsychological evaluation of children with developmental dyspraxia or consecutive to a preterm birth]. AB - OBJECTIVES: The aim of the present study was to compare and contrast developmental dyspraxia and dyspraxia following preterm birth. MATERIAL: Three different domains (attention-executive function, sensorimotor and visuospatial functions) were evaluated using Nepsy battery tests and the Purdue pegboard test. METHOD: The results of a neuropsychological evaluation of 32 children with developmental dyspraxia (the DVLT group) were compared with those of 16 children with dyspraxia following preterm birth (the AP group). The population inclusion and exclusion criteria were set according to Gerard's (Gerard C-L. Le concept de dyspraxie. In: Gerard C-L, Brun V, editors. Les dyspraxies de l'enfant. Paris: Masson; 2005. p. 15-24.) definition of dyspraxia. RESULTS: The two groups had similar visuospatial performance. However, differences between the DVLT and AP children appeared in a test of attention-executive function (visual attention, specifically) and in some sensorimotor tests, with better performance by the DVLT group in all situations. CONCLUSION: The difficulties exhibited by preterm-born children (poor upper limb motor control, more severe dyspraxia and the presence of oculomotor impairments) could explain the latter's poor performance relative to children with developmental dyspraxia. Indeed, oculomotor impairments are especially likely to impair performance in visual attention and visuomotor accurary tests - the two situations in which preterm-born children performed worse. PMID- 18395288 TI - Critical role of CD44 in hepatotoxin-mediated liver injury. AB - BACKGROUND/AIMS: Blocking of adhesion molecules is considered to be one of the therapeutic strategies inflammatory diseases, although it remains unclear whether this strategy is beneficial. METHODS: We used CD44-deficient mice to assess whether inhibition of CD44 could control liver injury caused by carbon tetrachloride (CCl(4)). RESULTS: CD44-deficient mice exhibited suppressed liver inflammation during the early phase (within 6h) after CCl(4) injection due to reduced inflammatory cell infiltration and cytokine production, but showed severe liver inflammation with increased numbers of apoptotic hepatocytes at the late phase (after 12h). The induction of hepatocyte apoptosis was triggered by reduced NF-kappaB activity, which was induced by the low inflammatory cytokine concentrations. Furthermore, macrophages contributed to the induction of hepatocyte apoptosis, since neutralization by an anti-CD11b antibody significantly protected against hepatocyte apoptosis. Finally, we found that blocking of MIP-2 and TNF-alpha reduced hepatocyte apoptosis with decreased numbers of intrahepatic leukocytes and reduced inflammatory cytokine production. CONCLUSIONS: These findings suggest that targeting of CD44 as a therapeutic approach for inflammatory liver diseases may require caution for particular immune systems in the liver. PMID- 18395287 TI - Fructose consumption as a risk factor for non-alcoholic fatty liver disease. AB - BACKGROUND/AIMS: While the rise in non-alcoholic fatty liver disease (NAFLD) parallels the increase in obesity and diabetes, a significant increase in dietary fructose consumption in industrialized countries has also occurred. The increased consumption of high fructose corn syrup, primarily in the form of soft drinks, is linked with complications of the insulin resistance syndrome. Furthermore, the hepatic metabolism of fructose favors de novo lipogenesis and ATP depletion. We hypothesize that increased fructose consumption contributes to the development of NAFLD. METHODS: A dietary history and paired serum and liver tissue were obtained from patients with evidence of biopsy-proven NAFLD (n=49) without cirrhosis and controls (n=24) matched for gender, age (+/-5 years), and body mass index (+/-3 points). RESULTS: Consumption of fructose in patients with NAFLD was nearly 2- to 3-fold higher than controls [365 kcal vs 170 kcal (p<0.05)]. In patients with NAFLD (n=6), hepatic mRNA expression of fructokinase (KHK), an important enzyme for fructose metabolism, and fatty acid synthase, an important enzyme for lipogenesis were increased (p=0.04 and p=0.02, respectively). In an AML hepatocyte cell line, fructose resulted in dose-dependent increase in KHK protein and activity. CONCLUSIONS: The pathogenic mechanism underlying the development of NAFLD may be associated with excessive dietary fructose consumption. PMID- 18395290 TI - Overview of the clinical application of regenerative medicine products in Japan. AB - OBJECTIVE: To identify barriers to the clinical application of regenerative medicine products (RMPs) in Japan. METHODS: Current Japanese regulatory systems and guidelines were compared with those of the United States (US) and the European Union (EU). A questionnaire was administered to representatives from 23 Japanese companies and 10 research institutes, and an in-person semi-structured interview was conducted with representatives from 10 companies that develop RMP. RESULTS: We found that Japan, the US and the EU have similar pre-clinical safety guideline frameworks relating to RMP. However, differences exist between these countries with respect to their review and approval systems and the implementation of guidelines, and these represent major barriers to the clinical application of RMP in Japan. Most companies studied are facing regulatory hurdles such as stringent review processes and regulatory guidelines that do not provide detailed practical examples of the pre-clinical quality and safety data required. CONCLUSIONS: These results suggest that effective regulatory infrastructure including regulatory systems, guidelines, and communication channels between product developers and regulatory bodies are essential for the prompt clinical application of RMP in Japan. PMID- 18395289 TI - Antibiotics protect against fructose-induced hepatic lipid accumulation in mice: role of endotoxin. AB - BACKGROUND/AIMS: Consumption of refined carbohydrates in soft drinks has been postulated to be a key factor in the development of non-alcoholic fatty liver disease (NAFLD). The aim of the present study was to test the effects of ad libitum access to different sugars consumed in drinking water on hepatic fat accumulation. METHODS: For 8 weeks, C57BL/J6 mice had free access to solutions containing 30% glucose, fructose, sucrose, or water sweetened with artificial sweetener (AS) or plain water. Body weight, caloric intake, hepatic steatosis and lipid peroxidation were assessed. RESULTS: Total caloric intake and weight gain were highest in mice exposed to glucose. In contrast, hepatic lipid accumulation was significantly higher in mice consuming fructose compared to all other groups. Moreover, endotoxin levels in portal blood and lipid peroxidation as well as TNFalpha expression were significantly higher in fructose fed mice than in all other groups. Concomitant treatment of fructose fed mice with antibiotics (e.g., polymyxin B and neomycin) markedly reduced hepatic lipid accumulation in fructose fed mice. CONCLUSIONS: These data support the hypothesis that high fructose consumption may not only lead to liver damage through overfeeding but also may be directly pro-inflammatory by increasing intestinal translocation of endotoxin. PMID- 18395292 TI - let-7 microRNA expression and the distinction between nonmucinous and mucinous bronchioloalveolar carcinomas. PMID- 18395291 TI - Multilayered polyelectrolyte assemblies as platforms for the delivery of DNA and other nucleic acid-based therapeutics. AB - Materials that provide spatial and temporal control over the delivery of DNA and other nucleic acid-based agents from surfaces play important roles in the development of localized gene-based therapies. This review focuses on a relatively new approach to the immobilization and release of DNA from surfaces: methods based on the layer-by-layer assembly of thin multilayered films (or polyelectrolyte multilayers, PEMs). Layer-by-layer methods provide convenient, nanometer-scale control over the incorporation of DNA, RNA, and oligonucleotide constructs into thin polyelectrolyte films. Provided that these assemblies can be designed in ways that permit controlled film disassembly under physiological conditions, this approach can contribute new methods for spatial and/or temporal control over the delivery of nucleic acid-based therapeutics in vitro and in vivo. We describe applications of layer-by-layer assembly to the fabrication of DNA-containing films that can be used to provide control over the release of plasmid DNA from the surfaces of macroscopic objects and promote surface-mediated cell transfection. We also highlight the application of these methods to the coating of colloidal substrates and the fabrication of hollow micrometer-scale capsules that can be used to encapsulate and control the release or delivery of DNA and oligonucleotides. Current challenges, gaps in knowledge, and new opportunities for the development of these methods in the general area of gene delivery are discussed. PMID- 18395293 TI - Effects of gibberellin A(3) on growth and microcystin production in Microcystis aeruginosa (cyanophyta). AB - Environmental factors that affect the growth and microcystin production of microcystis have received worldwide attention because of the hazards microcystin poses to environmental safety and public health. Nevertheless, the effects of organic anthropogenic pollution on microcystis are rarely discussed. Gibberellin A(3) (GA(3)) is a vegetable hormone widely used in agriculture and horticulture that can contaminate water as an anthropogenic pollutant. Because of its common occurrence, we studied the effects of GA(3) on growth and microcystin production of Microcystis aeruginosa (M. aeruginosa) PCC7806 with different concentrations (0.001-25mg/L) in batch culture. The control was obtained without gibberellin under the same culture conditions. Growth, estimated by dry weight and cell number, increased after the GA(3) treatment. GA(3) increased the amounts of chlorophyll a, phycocyanin and cellular-soluble protein in the cells of M. aeruginosa PCC7806, but decreased the accumulation of water-soluble carbohydrates. In addition, GA(3) was observed to affect nitrogen absorption of the test algae, but to have no effect on the absorption of phosphorus. The amount of microcystin measured by enzyme-linked immunosorbent assay (ELISA) increased in GA(3) treatment groups, but the stimulatory effects were different in different culture phases. It is suggested that GA(3) increases M. aeruginosa growth by stimulating its absorbance of nitrogen and increasing its ability to use carbohydrates, accordingly increasing cellular pigments and thus finally inducing accumulation of protein and microcystin. PMID- 18395294 TI - Alefacept for moderate to severe atopic dermatitis: a pilot study in adults. AB - BACKGROUND: Atopic dermatitis is a common inflammatory skin condition with acute and chronic phases showing a prevalence of memory T cells. Alefacept is a fully human LFA-3/IgG1 fusion protein that inhibits T-cell activation and selectively reduces memory T cells, which may prove to be effective in the treatment of atopic dermatitis. OBJECTIVE: We sought to evaluate clinical response of alefacept intramuscular (IM) injection for 16 weeks in adults with atopic dermatitis. METHODS: This was an open-label study of a 16-week treatment regimen of alefacept IM injection in adults with moderate to severe inflammatory atopic dermatitis. Patients received alefacept (30 mg IM) weekly for the first 8 weeks. At week 9, patients who did not achieve a 50% reduction in their Eczema Area Severity Index (EASI) score continued on alefacept (30 mg IM) weekly; those patients with a 50% reduction in their EASI (EASI 50) score or higher had their weekly dose decreased (15 mg IM) for the remaining 8 weeks. RESULTS: Nine patients with moderate to severe atopic dermatitis were enrolled and treated. At the primary end point, week 18, 1 patient achieved EASI 50 score and 1 patient achieved EASI 90 score; 4 patients had a decrease in EASI score of less than 50%, 1 patient had an increase in EASI score, and 2 patients withdrew early before the primary end point because of worsening disease. A Physician Global Assessment score of mild was achieved in 2 patients and 1 patient achieved a Physician Global Assessment score of almost clear. Minimal pruritus was reported by 3 patients and 1 patient reported no pruritus. The 16-week course of alefacept was well tolerated. LIMITATIONS: The study was inherently limited by its small sample size, concomitant use of antihistamines, and open-label design, which increases the likelihood of observer and self-assessment bias. CONCLUSION: The treatment regimen of alefacept for 16 weeks was well tolerated by our patients. Although, in this study, only 2 of the 9 patients with atopic dermatitis responded to treatment with alefacept, the study was inherently limited by the small sample size. Additional studies with a larger sample size, continued weekly use, or concomitant use of ultraviolet-B light therapy may be warranted to evaluate the possibility of alefacept as a therapy for patients with chronic atopic dermatitis. PMID- 18395295 TI - Knowledge and attitude toward Crimean-Congo haemorrhagic fever in occupationally at-risk Iranian healthcare workers. AB - We assessed the knowledge and attitude (K&A) toward Crimean-Congo haemorrhagic fever (CCHF) of occupationally at-risk healthcare workers (HCWs). A cross sectional survey was performed in three referral hospitals in the Systan Baluchestan and Isfahan provinces of Iran where CCHF is highly endemic. In all, 191/209 eligible HCWs were enrolled (response rate: 93%). All but 11 (5.8%) had heard of CCHF. The mean K&A scores of the respondents were 50.34% and 79.25%, respectively. The correlation between K&A was significant (correlation coefficient: 0.542; P<0.001). Being a physician, working in Isfahan (versus the relatively deprived Systan-Baluchestan) and relying on academic material rather than local media were independent factors significantly associated with more knowledge; higher education and laboratory staff with better attitude were also significant factors. Although HCWs showed the best K&A for preventive measures, only 44% wore gloves and masks for contact with CCHF patients and 22% failed to observe any safety measure. Those with a history of percutaneous contact (6.3%) had significantly lower knowledge scores (P=0.047). There is a need to establish professional education campaigns in highly endemic deprived areas in order to improve physicians' attitudes, encourage nurses' use of academic materials and increase the knowledge of less-educated HCWs. PMID- 18395296 TI - Results from the Scottish National HAI Prevalence Survey. AB - A national point prevalence survey was undertaken over the period of one calendar year in Scotland from October 2005 to October 2006. The prevalence of healthcare associated infection (HAI) was 9.5% in acute hospitals and 7.3% in non-acute hospitals. The highest prevalence of HAI in acute hospital inpatients was found in the following specialties: care of the elderly (11.9%), surgery (11.2%), medicine (9.6%) and orthopaedics (9.2%). The lowest prevalence was found in obstetrics (0.9%). The most common types of HAI in acute hospital inpatients were: urinary tract infections (17.9% of all HAI), surgical site infections (15.9%) and gastrointestinal infections (15.4%). In non-acute hospitals one in ten inpatients in two specialties (combined) - medicine (11.4%) and care of the elderly (7.8%) - was found to have HAI, and one in 20 inpatients in psychiatry (5.0%) had HAI. In non-acute hospital patients, urinary tract infections were frequent (28.1% of all HAI) and similarly skin and soft tissue infection (26.8% of all HAI). When combined, these two HAI types affected 4% of all the inpatients in non-acute hospitals. This is the first survey of its kind in Scotland and describes the burden of HAI at a national level. PMID- 18395297 TI - Synthesis, antitumor evaluation and DNA binding studies of novel amidino benzimidazolyl substituted derivatives of furyl-phenyl- and thienyl-phenyl acrylates, naphthofurans and naphthothiophenes. AB - A series of amidino-substituted benzimidazoles, related to furyl-phenyl- and thienyl-phenyl-acrylates, naphthofurans and naphthothiophenes were prepared, their antitumor evaluation and interactions with ct-DNA have been investigated. All tested compounds show differential and strong antitumor activity without apparent difference depending on their structures. Interestingly, the MCF-7 tumor cell line is highly sensitive to all compounds. Compounds 6-9 showed noticeable selectivity in regard to normal fibroblasts (WI 38). Compounds 4-9 interact with ct-DNA by more binding modes, whose mutual distribution is dependent on the compound/DNA ratio. The "acyclic"4-6 and "cyclic" compound 7 interact mostly within the minor groove of DNA, although partial intercalation of 6 and 7 cannot be excluded. The "cyclic" compounds 8 and 9 intercalate between DNA base pairs at high excess of DNA over compounds. PMID- 18395298 TI - Pharmacophore based discovery of potential antimalarial agent targeting haem detoxification pathway. AB - Pharmacophore hypotheses were generated from molecules having putative antimalarial activities targeting haem detoxification pathway of malarial parasite. A training set consisting of 33 compounds, whose activities were evaluated for haem polymerization inhibition and against chloroquine resistant (K1) strain of Plasmodium falciparum, was optimized to generate hypotheses. The hypothesis showing optimum correlation between actual and estimated activities was validated by Fischer's randomization test at 95% confidence level and used as a model to screen our in-house compound database. Nicotinic acid [trans-3-(4 ethoxy-3-methoxy-phenyl)-1-(4-hydroxy-phenyl)-allylidene]-hydrazide (ALH5) was obtained as a hit. The compound was synthesized and evaluated against chloroquine sensitive (MRC-02) and resistant (RKL9) strains of malarial parasite P. falciparum. The compound showed antimalarial activity in nanomolar range and was found comparable with chloroquine. PMID- 18395299 TI - 1,3,4-Oxadiazole/thiadiazole and 1,2,4-triazole derivatives of biphenyl-4-yloxy acetic acid: synthesis and preliminary evaluation of biological properties. AB - A series of 1,3,4-oxadiazole/thiadiazole and 1,2,4-triazole derivatives of biphenyl-4-yloxy acetic acid were synthesized in order to obtain new compounds with potential anti-inflammatory activity, analgesic activity and lower ulcerogenic potential. All compounds were evaluated for their anti-inflammatory activity by the carrageenan induced rat paw edema test method. The compounds possessing potent anti-inflammatory activity were further tested for their analgesic, ulcerogenic and antioxidant activities. Out of all tested compounds, the compounds 3, 7, 17 and 20, showed significant reduction in rat paw edema induced by carrageenan treatment. These compounds showed significant analgesic effect and at an equimolar oral doses relative to flurbiprofen were also found to be non-gastrotoxic in rats. Compound 17 was evaluated as the lead compound having more anti-inflammatory activity (81.81%) than the reference drug (79.54%), low ulcerogenic potential and protective effect on lipid peroxidation. PMID- 18395300 TI - Synthesis and biological evaluation of aminomethylphenol derivatives as inhibitors of the murine GABA transporters mGAT1-mGAT4. AB - A series of N-substituted aminomethylphenol derivatives was synthesized by reductive amination. To study the inhibitory potency of the target compounds at the murine GABA transporters (mGAT1-mGAT4), a [(3)H]GABA uptake test system in a 96-well format based on HEK cells stably expressing mGAT1-mGAT4 was established and validated. Inhibitory potencies at mGAT1-mGAT4 in the micromolar range and a slight subtype selectivity for mGAT3 were observed for the synthesized aminomethylphenol derivatives. Among the compounds investigated 5-n dodecylaminomethyl-2-methoxyphenol (21) was found to be most potent with an IC(50) value at mGAT3 of about 3muM. PMID- 18395301 TI - [Abdominal pain in essential thrombocythemia]. PMID- 18395302 TI - [Appropriateness of hospitalisation use: about the difference between productivity and health efficiency]. PMID- 18395303 TI - [Spontaneous low cerebospinal fluid pressure syndrome. A case report and literature review]. AB - INTRODUCTION: Spontaneous low cerebrospinal fluid pressure syndrome is a spontaneous intracranial hypotension pressure due to a cerebrospinal fluid leak without any known dural effraction. It is clinically characterised by postural headaches relieved by supine position. We report a 38-year-old patient with this syndrome and review the literature. EXEGESIS: The diagnosis is sometimes difficult in atypical presentation of the syndrome and can lead to incapacitating chronic headache and rarely to complications. Cerebral magnetic resonance imaging has dramatically improved identification, diagnosis and management of this syndrome. Treatment is mainly based on blood patch realisation. Cerebrospinal fluid leak probably due to a spontaneous defect in the dural mater is suspected to be the main mechanism of this syndrome without any history of lumbar puncture or penetrating trauma. CONCLUSION: Early diagnosis, often easy on the basis of clinical characteristics of the headache may avoid complications. PMID- 18395304 TI - [Potential nosocomial disseminated infection due to Nocardia asteroides after a prosthesis insertion in an immunocompetent patient]. AB - Nocardia infections are rare and usually occurred in immunocompromised patients with systemic dissemination from a lung infection. We report a case of an immunocompetent patient in whom Nocardia asteroides had cause psoas and cerebral abcess without pulmonary infection, a short period after a hip prosthesis insertion. The clinical history is highly suggestive of a hospital-acquired infection. PMID- 18395305 TI - Immunogenicity and safety of three consecutive lots of a new preservative-free inactivated hepatitis A vaccine (Healive): a double-blind, randomized and controlled trial. AB - Immunization is considered as the most effective way for the prophylaxis of hepatitis A virus (HAV) infection. This study aimed to evaluate the immunogenicity and safety of three consecutive lots of a new preservative-free inactivated hepatitis A vaccine (Healive) in healthy children. A double-blind, randomized and controlled clinical trial was conducted in healthy volunteers aged from 1 to 8 years. Total 400 subjects were enrolled and assigned into four groups, receiving one of the three lots of Healive or an established control vaccine. The vaccination was two-dose regimen with 6 months apart. Anti-HAV titers were determined at the 1st, 6th and 7th month. The results showed that Healive was highly immunogenic in children with 100% seroconversion rate (SR) and 3237-3814 mIU/ml geometry mean titer (GMT) 1 month after the second dose. The immunogenicity of Healive was statistically higher than that of the control vaccine with respect to GMT and SR (P=0.037 to P<0.001). Both Healive and control vaccine were well tolerated with 1-5% incidence of overall adverse reactions (P>0.298). Severe adverse reaction was not reported. Both SRs (1, 6 and 7 months) and GMTs (1 and 7 months) in subjects receiving one of the three consecutive lots of Healive had not statistical difference (P=0.114-0.710), suggesting that Healive was well consistent. The immune responses in younger children (1-3 years) and older children (4-8 years) were similar to each other (P=0.187-0.963). The present study indicated that Healive was greatly consistent between production lots, well tolerated and highly immunogenic in children, which made the preservative-free inactivated hepatitis A vaccine well suitable for inclusion in the routine programme of children vaccination. PMID- 18395306 TI - Multi-antigen vaccines based on complex adenovirus vectors induce protective immune responses against H5N1 avian influenza viruses. AB - There are legitimate concerns that the highly pathogenic H5N1 avian influenza virus could adapt for human-to-human transmission and cause a pandemic similar to the 1918 "Spanish flu" that killed 50 million people worldwide. We have developed pandemic influenza vaccines by incorporating multiple antigens from both avian and Spanish influenza viruses into complex recombinant adenovirus vectors. In vaccinated mice, these vaccines induced strong humoral and cellular immune responses against pandemic influenza virus antigens, and protected vaccinated mice against lethal H5N1 virus challenge. These results indicate that this multi antigen, broadly protective vaccine may serve as a safer and more effective approach than traditional methods for development of a pandemic influenza vaccine. PMID- 18395307 TI - Superiority of needle-free transdermal plasmid delivery for the induction of antigen-specific IFNgamma T cell responses in the dog. AB - Although successful needle-free DNA vaccination has been described on several occasions, the true benefit of this delivery technology over needle-based injections for DNA vaccination of dogs has not yet been documented. We conducted a side-by-side comparison of needle-free transdermal plasmid delivery vs. intramuscular vs. intradermal needle-based delivery of the same plasmid in dogs. Our data confirmed the importance of the route of plasmid delivery and further established the unique potential of needle-free transdermal plasmid delivery to elicit strong antigen-specific, hTyr-specific IFNgamma T in the dog. Further, this study demonstrated that properly enabled DNA vaccination has the potential to trigger very significant cell-based immune responses in dogs, establishing needle-free transdermal plasmid delivery as a critical technology for successful immunotherapy of cancer and/or chronic infectious diseases in companion animal medicine. PMID- 18395308 TI - Decision-making in environmental radiation protection: using the ERICA Integrated Approach. AB - The ERICA Integrated Approach and its associated tool and databases provide a method by which the likely impact of radioactive discharges on the environment can be evaluated, see Fig. 1. The various factors, which should be taken into account when making decisions both during and after an assessment has been made, are discussed for each stage in the assessment process for a hypothetical case study. The assessment will demonstrate the issues associated with the decision making process at Tiers 1 and 2 within the ERICA Tool and how they may vary. The case study, set in England, evaluates the environmental impact of radioactive substances released under authorisation in response primarily to conservation legislation, because of the need to demonstrate that no adverse impacts will occur on Natura 2000 sites as a result of the release of an authorised substance. PMID- 18395309 TI - Are pregnant women of non-Northern European descent more anaemic than women of Northern European descent? A study into the prevalence of anaemia in pregnant women in Amsterdam. AB - OBJECTIVES: to investigate the prevalence of anaemia in pregnancy according to the cut-off points used in the national clinical guideline 'Anaemia in Primary Care Midwifery Practice', and to investigate a possible difference in prevalence between pregnant women of Northern European descent compared with women of non Northern European descent. The study also investigated whether any difference in prevalence resulted in a higher risk of a transferred home delivery, thus reducing the chance for women to choose the place of birth. DESIGN: retrospective cohort study. SETTING: primary care midwifery practices in a highly urbanised area in Amsterdam, the Netherlands. PARTICIPANTS: all patients in three midwifery practices over 1 year were selected provided that they met the inclusion and exclusion criteria. The practices were selected on the basis of urbanisation, resulting in an adequate ethnic population which was comparable with the ethnic mix in other similarly urbanised areas in the Netherlands. This resulted in a study population of 828 pregnant women of Northern European and non-Northern European descent with low-risk singleton pregnancies during 2003. ANALYSIS: mean haemoglobin level and anaemia rate were computed for the total study population as well as separate ethnic groups. Differences between groups were determined using chi(2) tests, and effect sizes were expressed in relative risks including 95% confidence intervals (CI). FINDINGS: the prevalence of anaemia in pregnancy was 3.4% (n = 28) at booking (first visit) and 2.7% (n = 22) at approximately 30 weeks of gestation. The relative risk of anaemia at booking was 5.9 (95% CI 2.1 16.7) for pregnant women of non-Northern European descent compared with those of Northern European descent. The relative risk of anaemia at approximately 30 weeks of gestation was 22 (95% CI 3-162) for pregnant women of non-Northern European descent compared with those of Northern European descent. The incidence of transferred home delivery was 3.1% in the study population. The relative risk for transferred home delivery was 24.1 (95% CI 3.3-176.7) for pregnant women of non Northern European descent compared with those of Northern European descent. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: pregnant women of non-Northern European descent are a specific group in terms of anaemia and are at high risk of a low haemoglobin level for all cut-off values; they have a higher chance of becoming anaemic in pregnancy than women of Northern European descent. As a result, they have a higher risk of a transferred home delivery, taking away their opportunity to choose the place of birth. Haemoglobinopathies did not explain the higher risk of anaemia in pregnant women of non-Northern European descent. Being of non-Northern European descent should be added as a risk factor for anaemia in pregnancy until more is known about the background of this risk factor. PMID- 18395310 TI - Influences of hip external rotation strength on knee mechanics during single-leg drop landings in females. AB - BACKGROUND: To determine the influence of hip external rotation strength on kinematic and kinetic variables during single-leg drop landings. METHODS: Females were divided into strong and weak groups based on isometric hip external rotation strength and lower extremity flexion-extension and varus-valgus kinematics and kinetics were evaluated during single-legged 40 cm drop landings. FINDINGS: Hip external rotation strength had no effect on sagittal or frontal plane angular motion throughout the landing cycle at the hip and knee. The strong group generated a statistically significant decrease in the peak vertical ground reaction force and external knee flexor moment. The weak group produced a greater external knee adduction moment, net knee anterior shear joint reaction force, and a greater hip external adduction moment. INTERPRETATION: Subjects with greater hip and quadriceps/hamstring strength exhibited a significant decrease in the vertical ground reaction force and external knee adduction and flexor moments. Hip and knee strength appear to relate to differences in high risk landing strategies. PMID- 18395311 TI - Groundwater-soil-crop relationship with respect to arsenic contamination in farming villages of Bangladesh--a preliminary study. AB - To clarify the groundwater-soil-crop relationship with respect to arsenic (As) contamination, As concentration was measured in tubewell (TW) water, surface soil from farmyards and paddy fields, and fresh taro (Colocasia esculenta) leaves from farmyards in the farming villages of Bangladesh. The As concentration in TW water from farmyards was at least four times higher than the Bangladesh drinking water standard, and the concentration in fresh taro leaves was equal to or higher than those reported previously for leafy vegetables in Bangladesh. As concentration of surface soils in both farmyards and paddy fields was positively correlated with that of the TW water. Further, the concentration in surface soil was positively correlated with levels in fresh taro leaves in the farmyard. This study, therefore, clarified the groundwater-soil-crop relationship in farmyards and the relationship between groundwater-soil in paddy fields to assess the extent of As contamination in Bangladeshi villages. PMID- 18395312 TI - Monitoring nitrogen accumulation in mosses in central European forests. AB - In order to assess whether nitrogen (N) loads in mosses reflect different land uses, 143 sites in North Rhine-Westphalia, the Weser-Ems Region and the Euro Region Nissa were sampled between 2000 and 2005. The data were analysed statistically with available surface information on land use and forest conditions. N bioaccumulation in mosses in the Weser-Ems Region with high densities of agricultural land use and livestock exceeded the concentrations in the more industrialised Euro Region Nissa. In all three study areas agricultural and livestock spatial densities were found to be positively correlated with N bioaccumulation in mosses. In North Rhine-Westphalia, the N concentrations in mosses was also moderately correlated with N concentrations in leaves and needles of forest trees. The moss method proved useful to assess the spatial patterns of N bioaccumulation due to land use. PMID- 18395313 TI - Critical loads and their exceedances at intensive forest monitoring sites in Europe. AB - Intensive forest monitoring by means of harmonised methods has been conducted in Europe for more than a decade. Risks of atmospheric nitrogen and sulphur deposition are assessed by means of calculations of critical loads and their exceedances. In the present study throughfall and bulk deposition of nitrate (N NO(3)), ammonium (N-NH(4)) and sulphate (S-SO(4)) show marked spatial patterns and temporal trends. In the period of observation (1999-2004), sulphate deposition on intensive monitoring plots decreased by about one quarter. This is in line with the reduction of S deposition by 70% since 1981 in Europe as a result of successful air pollution control politics under the Convention on Long range Transboundary Air Pollution (CLRTAP). However, sulphate and especially nitrate and ammonium deposition were found to still exceed critical loads at many forest sites, indicating a continued need for further implementation of air pollution abatement strategies. PMID- 18395314 TI - [Jules Guerin (1801-1885) and wound pneumatic occlusion, a precursor of V.A.C.((R))]. AB - Vacuum-Assisted Closure System((R)) (V.A.C.((R))), tested and introduced by MJ Moryk- was and LC Argenta in 1977, is a wide-spread procedure used for the treatment of defects and chronic wounds. In the middle of the 19th century, a French surgeon tried hopelessly to convince the medical authorities by that time of the interest of a similar method. In 1844, Jules Guerin (1801-1885) imagined a process and described the technique and the results in the prestigious Academy of Medicine and Sciences in Paris in 1866 and 1867. Jules Guerin proposed to treat chronic wounds and defects by what he called "pneumatic occlusion". He proposed an apparatus with three parts: a process dressing applied to be wound a rubber occlusive membrane linked to an empty bell with a manometer. Jules Guerin presented his method with the indications, severed times at the Academy of Medicine. Results were detailed on complex wounds or amputations strumps. He explained with detail the action of the pneumatic occlusion but without taking on to account the role microorganisms and the absence of hygiene in infections. Unfortunately for him, his presentations were contemporary to the works of Pasteur and Lister. All surgeons privileged the use of antiseptics on wounds. The works of Jules Guerin were forgotten. PMID- 18395315 TI - Effect of curing mode on the polymerization characteristics of dual-cured resin cement systems. AB - OBJECTIVES: To evaluate the effects of different curing conditions on the degree of conversion (DC) of dual-cured cementing systems [combination of bonding agent (BA) and resin cement (RC)] using infrared spectroscopy. METHODS: Four fourth generation products [Scotchbond Multipurpose Plus/RelyX (3M ESPE), Optibond/Nexus 2 (Kerr), All Bond2/Duolink (Bisco), and Bond-It!/Lute-It! (Pentron)], and three fifth generation materials [Bond1/Lute-It! (Pentron), Prime&Bond NT Dual Cure/Calibra (Dentsply), and Optibond Solo Dual Cure/Nexus 2 (Kerr)] were applied to the surface of a horizontal attenuated-total-reflectance unit, and were polymerized using one of four conditions: self-cure (SC), direct light exposure through glass slide (DLE, XL3000/3M ESPE) or through pre-cured resin discs (shades A2;A4/2mm thick/Z250/3M ESPE). Infrared spectra of the uncured cementing systems were recorded immediately after application to the ATR, after the system was light-cured or left to self-cure, and spectra were obtained 5 and 10 min later. DC was calculated using standard techniques of observing changes in aliphatic-to-aromatic peak ratios pre- and post-curing. Data (n=5) were analyzed by two-way repeated measures ANOVA and Tukey's test (p=0.05). RESULTS: Changes in aliphatic-to-aromatic peak ratios before and after placing RC onto the BA demonstrated that a combined layer was created. All groups exhibited higher DC after 10 min than after 5 min, except the DLE group of Bond-it!/Lute-it!. No significant differences in DC were observed among light-activated groups regardless of the resin disc shade in three of the four fourth generation cementing systems. The SC groups exhibited lower DC than the DLE groups for both fourth and fifth generation products either after 5 or 10 min. CONCLUSION: The chemistry of the bonding interface changed when RCs were applied to uncured BAs. The presence of an indirect restoration can decrease the DC of some cementing systems and the self-curing mode leads to lower DC than the light-activating one. PMID- 18395316 TI - Listeners modulate temporally selective attention during natural speech processing. AB - Spatially selective attention allows for the preferential processing of relevant stimuli when more information than can be processed in detail is presented simultaneously at distinct locations. Temporally selective attention may serve a similar function during speech perception by allowing listeners to allocate attentional resources to time windows that contain highly relevant acoustic information. To test this hypothesis, event-related potentials were compared in response to attention probes presented in six conditions during a narrative: concurrently with word onsets, beginning 50 and 100 ms before and after word onsets, and at random control intervals. Times for probe presentation were selected such that the acoustic environments of the narrative were matched for all conditions. Linguistic attention probes presented at and immediately following word onsets elicited larger amplitude N1s than control probes over medial and anterior regions. These results indicate that native speakers selectively process sounds presented at specific times during normal speech perception. PMID- 18395318 TI - Salts enhance both protein stability and amyloid formation of an immunoglobulin light chain. AB - Amyloid fibrils are associated with sulfated glycosaminoglycans in the extracellular matrix. The presence of sulfated glycosaminoglycans is known to promote amyloid formation in vitro and in vivo, with the sulfate groups playing a role in this process. In order to understand the role that sulfate plays in amyloid formation, we have studied the effect of salts from the Hofmeister series on the protein structure, stability and amyloid formation of an amyloidogenic light chain protein, AL-12. We have been able to show for the first time a direct correlation between protein stability and amyloid formation enhancement by salts from the Hofmeister series, where SO(4)(2-) conferred the most protein stability and enhancement of amyloid formation. Our study emphasizes the importance of the effect of ions in the protein bound water properties and downplays the role of specific interactions between the protein and ions. PMID- 18395319 TI - Stem cells for regenerative medicine--biological attributes and clinical application. AB - The place of stem cells in the therapeutic armamentarium is currently subject to intense scrutiny. One issue is the nature of the stem cells themselves, and their relationship to better-known stem cells, such as hematopoietic stem cells. Here it is argued that the stem cells we propose for therapeutic application may be more closely related to embryonic cells than are hematopoietic stem cells. A second issue is how the stem cells may best be manipulated for therapeutic use; this includes stem cell purification, amplification, fate selection, and suitability for clinical use. PMID- 18395320 TI - Predictive models of turbidity and water depth in the Donana marshes using Landsat TM and ETM+ images. AB - We have used Landsat-5 TM and Landsat-7 ETM+ images together with simultaneous ground-truth data at sample points in the Donana marshes to predict water turbidity and depth from band reflectance using Generalized Additive Models. We have point samples for 12 different dates simultaneous with 7 Landsat-5 and 5 Landsat-7 overpasses. The best model for water turbidity in the marsh explained 38% of variance in ground-truth data and included as predictors band 3 (630-690 nm), band 5 (1550-1750 nm) and the ratio between bands 1 (450-520 nm) and 4 (760 900 nm). Water turbidity is easier to predict for water bodies like the Guadalquivir River and artificial ponds that are deep and not affected by bottom soil reflectance and aquatic vegetation. For the latter, a simple model using band 3 reflectance explains 78.6% of the variance. Water depth is easier to predict than turbidity. The best model for water depth in the marsh explains 78% of the variance and includes as predictors band 1, band 5, the ratio between band 2 (520-600 nm) and band 4, and bottom soil reflectance in band 4 in September, when the marsh is dry. The water turbidity and water depth models have been developed in order to reconstruct historical changes in Donana wetlands during the last 30 years using the Landsat satellite images time series. PMID- 18395317 TI - Neural phenotypes of common and rare genetic variants. AB - Neuroimaging methods offer a powerful way to bridge the gaps between genes, neurobiology and behavior. Such investigations may be further empowered by complementary strategies involving chromosomal abnormalities associated with particular neurobehavioral phenotypes, which can help to localize causative genes and better understand the genetics of complex traits in the general population. Here we review the evidence from studies using these convergent approaches to investigate genetic influences on brain structure: (1) studies of common genetic variations associated with particular neuroanatomic phenotypes, and (2) studies of possible 'genetic subtypes' of neuropsychiatric disorders with very high penetrance, with a focus on neuroimaging studies using novel computational brain mapping algorithms. Finally, we discuss the contribution of behavioral neurogenetics research to our understanding of the genetic basis of neuropsychiatric disorders in the broader population. PMID- 18395321 TI - Real-time 3-D contrast-enhanced transcranial ultrasound and aberration correction. AB - Contrast-enhanced (CE) transcranial ultrasound (US) and reconstructed 3-D transcranial ultrasound have shown advantages over traditional methods in a variety of cerebrovascular diseases. We present the results from a novel ultrasound technique, namely real-time 3-D contrast-enhanced transcranial ultrasound. Using real-time 3-D (RT3D) ultrasound and microbubble contrast agent, we scanned 17 healthy volunteers via a single temporal window and nine via the suboccipital window and report our detection rates for the major cerebral vessels. In 71% of subjects, both of our observers identified the ipsilateral circle of Willis from the temporal window, and in 59% we imaged the entire circle of Willis. From the suboccipital window, both observers detected the entire vertebrobasilar circulation in 22% of subjects, and in 44%, the basilar artery. After performing phase aberration correction on one subject, we were able to increase the diagnostic value of the scan, detecting a vessel not present in the uncorrected scan. These preliminary results suggest that RT3D CE transcranial US and RT3D CE transcranial US with phase aberration correction have the potential to greatly impact the field of neurosonology. PMID- 18395322 TI - Comparing the Gleason prostate biopsy and Gleason prostatectomy grading system: the Lahey Clinic Medical Center experience and an international meta-analysis. AB - BACKGROUND: The accuracy of the prostate biopsy Gleason grade to predict the prostatectomy Gleason grade varies tremendously in the literature. OBJECTIVES: Determine the accuracy and distribution of the prostate biopsy Gleason grade and prostatectomy Gleason grade at LCMC (Lahey Clinic Medical Center) and worldwide. DESIGN, SETTING, AND PARTICIPANTS: Participants included 2890 patients who had not received preoperative hormones, and for whom preoperative and postoperative Gleason sums were available. Participants underwent radical prostatectomy at LCMC, an academic referral center, from 1982-2007. Studies for the meta-analysis were selected from Medline: 1994-2007. Search criteria included keywords "Gleason," "biopsy," and "prostatectomy," >/=200 patients, and whether the biopsy and prostatectomy Gleason scores categorized into the predefined Gleason grades. The meta-analysis included 15 studies and the LCMC database for 14,839 total patients. MEASUREMENTS: Gleason scores 2-6, 7, and 8-10 were converted to low, moderate, and high grade, respectively. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value were calculated. The kappa statistic and chi-square were used to compare biopsy and prostatectomy grades. RESULTS AND LIMITATIONS: The percentage of patients in whom the prostatectomy grade was accurately predicted, upgraded, and downgraded was 58%, 36%, and 5% at LCMC and 63%, 30%, and 7% in the meta-analysis, respectively. The PPV for low-, moderate-, and high-grade cancer was 54%, 70%, and 60% for LCMC and 62%, 70%, and 50% for the meta-analysis, respectively. The sensitivity decreased with increasing Gleason grade (low, moderate, and high) for LCMC (91%, 38%, 28%) and the meta-analysis (90%, 40%, 33%), respectively. The distribution of low-, moderate-, and high-grade cancer on biopsy (69%, 25%, and 6%) and prostatectomy specimen (47%, 44%, and 9%) demonstrated only "fair" agreement (kappa, 0.37). CONCLUSIONS: Patients and practitioners need to be cognizant of significant upgrading for low-grade disease and the downgrading for high-grade disease. PMID- 18395323 TI - Editorial comment on: Metastin inhibits migration and invasion of renal cell carcinoma with overexpression of metastin receptor. PMID- 18395324 TI - Laparoscopic radical cystectomy-where are we really? PMID- 18395325 TI - Metastin inhibits migration and invasion of renal cell carcinoma with overexpression of metastin receptor. AB - BACKGROUND: Metastin, the final peptide of the KiSS-1 gene, has been proposed to suppress cell motility. OBJECTIVE: This study investigated whether renal cell carcinoma (RCC) tissue expresses metastin or its receptor, and clarified whether metastin can suppress migration and/or invasion and/or proliferation of RCC cells in vitro. DESIGN, SETTING, AND PARTICIPANTS: Twenty-five RCC samples were submitted. Fresh RCC tissues were prepared for real-time RT-PCR, and formalin fixed and paraffin-embedded tissues blocks were examined by immunohistochemistry. RCC cell lines Caki-1 and ACHN were supplied for cell migration, invasion, and proliferation assays. MEASUREMENTS: Real-time RT-PCR was performed by using Taq Man gene expression system. ENVISION system was used in immunohistochemistry. Wound-healing assay and matrigel assays were used to identify migration and invasion abilities of RCC cell lines. Cell Counting Kit-8 was applied to measure the cell proliferation. Cell morphology was examined under a META system. Statistical analysis was performed with SPSS15.0J. RESULTS AND LIMITATIONS: In twenty-five RCC samples, the mRNA level of metastin receptor was identified to be significantly higher than non-neoplastic renal cortex by real-time RT-PCR (p=0.011). Immunohistochemical study also detected metastin receptor protein in all RCC tumors. In vitro, this study showed that metastin inhibited migration and invasion of Caki-1 and ACHN cells. In contrast, it had no effects on cell proliferation. Metastin (10 micromol/l) induced excessive formation of focal adhesions and stress fibers in Caki-1 and ACHN cells; this phenomenon was inhibited by pretreating pharmacological Rho-kinase inhibitor (Y-27632) to those cells. CONCLUSION: This is the first report regarding overexpression of the metastin receptor hOT7T175 in human RCC. We demonstrate that metastin can inhibit migration and invasion of the RCC cell line, which is regulated by a Rho-kinase inhibitor. Metastin and its receptor are therefore probable targets for suppressing RCC. PMID- 18395326 TI - Comparable efficacy of once-daily versus on-demand vardenafil in men with mild-to moderate erectile dysfunction: findings of the RESTORE study. AB - BACKGROUND: Phosphodiesterase (type) 5 (PDE5) inhibitors are currently administered on demand for treatment of erectile dysfunction (ED). Once-daily dosing has been suggested to benefit patients. OBJECTIVE: To determine whether daily vardenafil use provides added clinical benefits to patients compared with on-demand dosing. DESIGN, SETTING, AND PARTICIPANTS: In this placebo-controlled, double-blind, multicentre parallel-group study, men with mild-to-moderate ED were randomised to 24 wk of treatment, followed by a 4-wk washout. INTERVENTION: Patients were randomised to receive once-daily vardenafil 10mg plus on-demand placebo for 12 or 24 wk, or once-daily placebo plus on-demand vardenafil 10mg for 24 wk. MEASUREMENTS: Primary efficacy variable was the between-group difference in change in International Index of Erectile Function-Erectile Function domain (IIEF-EF) score from baseline to end of washout. Secondary variables included change from baseline in proportion of positive respondents to Sexual Encounter Profile questions and in satisfaction with treatment as assessed with the Treatment Satisfaction Scale (TSS). RESULTS AND LIMITATIONS: LS mean changes from baseline in IIEF-EF scores were 2.02, 2.29, and 2.63 for vardenafil 12 wk once daily, 24 wk once daily, and 24 wk on demand, respectively. After washout, the trend was towards improved IIEF-EF scores in the on-demand group (20.58 [+/ 0.96]) versus both once-daily groups (12 wk, 19.88 [+/-0.93]; 24 wk, 20.11 [+/ 0.94]). Furthermore, there were no significant between-group differences in the percentage of patients with "normal" erectile function. TSS analyses demonstrated no significant differences between treatment groups. This study recruited patients with mild-to-moderate ED; therefore, the results may not be the same as in patients with severe ED. CONCLUSIONS: Once-daily vardenafil did not produce greater sustained effects on EF than on-demand vardenafil in men with mild-to moderate ED, suggesting that daily dosing of PDE5 inhibitors does not produce sustained clinical benefits beyond cessation of treatment above those observed with on-demand administration. PMID- 18395327 TI - Editorial comment on: comparing the Gleason prostate biopsy and Gleason prostatectomy grading system: the Lahey Clinic Medical Center experience and an international meta-analysis. PMID- 18395328 TI - Editorial comment on: comparing the Gleason prostate biopsy and Gleason prostatectomy grading system: the Lahey Clinic Medical Center experience and an international meta-analysis. PMID- 18395329 TI - Editorial comment on: Metastin inhibits migration and invasion of renal cell carcinoma with overexpression of metastin receptor. PMID- 18395330 TI - Editorial comment on: comparable efficacy of once-daily versus on-demand vardenafil in men with mild-to-moderate erectile dysfunction: findings of the RESTORE study. PMID- 18395331 TI - Skeletal stem cells: phenotype, biology and environmental niches informing tissue regeneration. AB - Advances in our knowledge of the biology of skeletal stem cells, together with an increased understanding of the regeneration of normal tissue offer exciting new therapeutic approaches in musculoskeletal repair. Skeletal stem cells from various adult tissues such as bone marrow can be identified and isolated based on their expression of a panel of markers associated with smooth muscle cells, pericytes and endothelial cells. Thus, skeletal stem cell-like populations within bone marrow may share a common perivascular stem cell niche within the microvascular network. To date, the environmental niche that nurtures and maintains the stromal stem cell at different anatomical sites remains poorly understood. However, an understanding of the osteogenic and perivascular niches will inform identification of the key growth factors, matrix constituents and physiological conditions that will enhance the ex vivo amplification and differentiation of osteogenic stem cells to mimic native tissue critical for tissue repair. This review will examine skeletal stem cell biology, the advances in our understanding of the skeletal and perivascular niche and interactions therein and the opportunities to harness that knowledge for musculoskeletal regeneration. PMID- 18395332 TI - Digital acroparesthesia caused by pontine infarction. PMID- 18395333 TI - Dietary agents for chemoprevention of prostate cancer. AB - Prostate cancer (CaP) is the leading cause of cancer-related deaths in American men, responsible for over 29,000 deaths in the year 2007. Chemoprevention is a plausible and cost-effective approach to reduce cancer morbidity and mortality through inhibition of precancerous events before the occurrence of clinical disease. Indeed, CaP is an ideal candidate disease for chemopreventive intervention as it is typically diagnosed in the elderly population with a relatively slower rate of growth and progression. The potential of dietary substances to act as chemopreventive agents against CaP is increasingly appreciated. Further, epidemiological studies have identified significant correlations between CaP incidence and dietary habits. It is hoped that, combining the knowledge based on agents with targets, we will be able to build an armamentarium of naturally occurring chemopreventive substances that could prevent or slow down the development and progression of CaP. In this review, we have summarized the findings from clinical and preclinical studies on dietary agents including green tea, pomegranate, lupeol, fisetin, and delphinidin that are currently being investigated in our laboratory for their chemopreventive potential against CaP. PMID- 18395334 TI - Removal of lead from aqueous solutions with a treated spent bleaching earth. AB - A spent bleaching earth from an edible oil refinery has been treated by impregnation with a normal sodium hydroxide solution followed by mild thermal treatment (100 degrees C). The obtained material (TSBE) was washed, dried and characterized by X-ray diffraction, FTIR, SEM, BET and thermal analysis. The clay structure was not apparently affected by the treatment and the impregnated organic matter was quantitatively removed. We have investigated the sorption of lead on this material, the spent bleaching earth (SBE) and the virgin bleaching earth (VBE). The kinetic results fit the pseudo second-order kinetic model and the Weber & Morris, intraparticle diffusion model. The pH had effect on the sorption efficiency. The sorption isotherms followed the Langmuir model for various sorbent concentrations with good values of determination coefficient. A comparison between the results obtained with this material and those of the literature highlighted a good removal capacity of the treated spent bleaching earth at low cost. PMID- 18395335 TI - Employment of polyurethane foam for the adsorption of Methylene Blue in aqueous medium. AB - This work presents a detailed study about the adsorption of Methylene Blue (MB) onto polyether type polyurethane foam (PUF). The adsorption process is based on the formation of a hydrophobic ionic-pair between cationic dye MB and dodecylsulfate anion (SDS), which present high affinity by PUF. Set-up employed in the study was built up by adjusting a 200mg cylinder of PUF to the arm of an overhead stirrer. The system was characterized in relation to equilibrium, kinetic and thermodynamic aspects and it was modeled by employing Langmuir and Freundlich isotherms. Obtained results showed that the ratio between SDS and MB concentrations plays an important role on the adsorption process. According to Langmuir isotherm, a maximum adsorption capacity of 7.20 x 10(-5) mol MBg(-1) was achieved when optimized operational conditions were employed. The adsorption rate seems to be regulated by an intraparticle diffusion mechanism. Adsorption process was spontaneous (negative DeltaG) at ambient temperature and presented an endothermic characteristic (positive DeltaH). Sequential extraction experiments were carried out by changing PUF plugs in 30 min time intervals and around 96% of the MB present in solution could be removed through consecutive extractions with six 200 mg PUF cylinders. PMID- 18395336 TI - Assessment of cement kiln dust (CKD) for stabilization/solidification (S/S) of arsenic contaminated soils. AB - A stabilization/solidification (S/S) process for arsenic (As) contaminated soils was evaluated using cement kiln dust (CKD). Laboratory-prepared slurries, made of either kaolinite or montmorillonite, and field soils spiked with either As(3+) or As(5+) were prepared and treated with CKD ranging from 10 to 25 wt%. Sodium arsenite and sodium arsenate at 0.1 wt% were used to simulate arsenite (As(3+)) and arsenate (As(5+)) source contamination in soils, respectively. The effectiveness of treatment was evaluated at curing periods of 1- and 7-days based on the toxicity characteristic leaching procedure (TCLP). As-CKD and As-clay-CKD slurries were also spiked at 10 wt% to evaluate As immobilization mechanism using X-ray powder diffraction (XRPD) analyses. Overall, the TCLP results showed that only the As(5+) concentrations in kaolinite amended with 25 wt% CKD after 1 day of curing were less than the TCLP regulatory limit of 5mg/L. Moreover, at 7 days of curing, all As(3+) and As(5+) concentrations obtained from kaolinite soils were less than the TCLP criteria. However, none of the CKD-amended montmorillonite samples satisfied the TCLP-As criteria at 7 days. Only field soil samples amended with 20 wt% CKD complied with the TCLP criteria within 1 day of curing, where the source contamination was As(5+). XRPD and scanning electron microscopy (SEM)-energy dispersive X-ray spectroscopy (EDX) results showed that Ca-As-O and NaCaAsO(4).7.5H(2)O were the primary phases responsible for As(3+) and As(5+) immobilization in the soils, respectively. PMID- 18395338 TI - Photocatalytic degradation of methyl tert-butyl ether in the gas-phase: a kinetic study. AB - Methyl tert-butyl ether (MTBE) is the basic oxygenated motor fuel additive in Europe and is included in volatile organic compounds (VOCs), which can produce photochemical oxidants. In the present study the gas-phase photocatalytic oxidation (PCO) of MTBE over illuminated titanium dioxide was carried out at ambient temperature in a plug flow reactor. The intermediates detected are mainly tert-butyl formate and acetone, while the final products are CO(2) and water. The system was sensitive to the oxygen concentration, for concentrations up to 15% (v/v). Moisture had a positive effect on the reaction, obtaining an optimum value near 45% relative humidity for 200ppmv MTBE initial concentration. A reaction scheme has been proposed for the interpretation of the experimental results and a kinetic study was conducted, using the Langmuir-Hinshelwood kinetics equation. The MTBE rate constant was 1.545 x 10(-6)Ms(-1)g(cat)(-1) for the reaction without moisture and 2.46 x 10(-6)Ms(-1)g(cat)(-1) for the reaction in the presence of moisture and the adsorption constant was 2.187 x 10(5)M(-1) independent of humidity. PMID- 18395337 TI - A novel process for the removal of aniline from wastewaters. AB - The aim of this research was to develop a solid regenerable catalytic adsorbent capable of removing aniline from aqueous solutions. A H-Beta zeolite was first loaded with copper in an ion-exchange process to enhance its catalytic activity. Experimental results indicated an aniline adsorption level of approximately 106 114 mg g(-1) for each of the unmodified H-Beta, the 0.5% (w/w) Cu-Beta or the 1.4 % (w/w) Cu-Beta zeolites. The adsorption processes followed the Langmuir model and the level of aniline adsorbed was largely unaffected by a change in temperature. Assessment of the aqueous stability of the exchanged copper on the Beta zeolites indicated minimum copper leaching in the range pH 5-11 thus providing a stable working pH range for both the 0.5% (w/w) and 1.4% (w/w) Cu Beta adsorbent materials. Catalytic oxidation studies on the adsorbed aniline indicated that the presence of copper in the zeolites significantly enhanced the degradation of aniline to predominantly carbon dioxide, water and nitrogen. Five successive adsorption/catalytic oxidation cycles did not diminish the aniline adsorption capacity of the copper loaded zeolites but there was a small loss in the efficacy of the catalytic oxidation of the adsorbed aniline by the end of the 5th cycle. PMID- 18395339 TI - Pilot study of temporal variations in lead bioaccessibility and chemical fractionation in some Chinese soils. AB - The effect of ageing, following the addition of approximately 400mgkg(-1) lead (Pb) as Pb(NO(3))(2), on Pb bioaccessibility was examined in five typical Chinese soils using a physiologically based extraction test. Sequential extraction was employed to identify the source fraction(s) of bioaccessible Pb in the soils. Pb bioaccessibility decreased exponentially to nearly steady levels in mildly acidic or alkali (pH 6.09-7.43) soils, for both gastric (69.91-71.75%) and small intestinal (7.53-9.63%) phases within the first 2-4 weeks and 1-2 months of incubation, respectively; however, it took only 1-2 weeks for strongly acidic ( approximately pH 4.5) soils to reach nearly steady levels of Pb bioaccessibility (73.01-74.46% and 10.30-10.98% in the gastric and small intestinal phases, respectively). In addition to the water-soluble and exchangeable fractions, the carbonate fraction of mildly acidic or alkali soils appeared to be a third main source of bioaccessible Pb in the small intestinal phase; however, bioaccessible Pb was likely to derive principally from Pb in the water-soluble and exchangeable fractions of strongly acidic soils. Bioaccessible Pb in the gastric phase appeared to derive from all the fractions in all five studied soils, even the residual fraction. PMID- 18395340 TI - Synthesis and metal ion binding properties of thiaaza crown macrocycles. AB - Two new macrocyclic ligands (3) and (4), containing nitrogen-sulfur donor atoms, were designed and synthesized in a multi-step reaction sequence. The macrocycles (3) and (4) were used in solvent extraction of metal picrates such as Ag(+), Hg(2+), Cd(2+), Zn(2+), Cu(2+), Ni(2+), Mn(2+), Co(2+), and Pb(2+) from aqueous phase to the organic phase. The metal picrate extractions were investigated at 25+/-0.1 degrees C by using UV-visible spectrometry. The extractability and selectivity of the mentioned metal picrates were evaluated according to the organic solvents. The values of the extraction constants (logK(ex)) and the complex compositions were determined for the extracted complexes. PMID- 18395341 TI - Reinvestigation of the effect of orexin A on catecholamine release from adrenal chromaffin cells. AB - Orexins have been shown to be implicated in the regulation of adrenal medulla functions. However, there are still inconsistent investigations on the effects of orexins on catecholamine release from chromaffin cells in varying species. In the present study, using the carbon-fiber amperometry, we investigated whether orexin A would stimulate catecholamine release from rat and mouse adrenal chromffin cells. Puff application of orexin A dose-dependently induced amperometric currents in the cultured rat chromaffin cells, which was completely blocked by the selective OX1R antagonist SB-334867 or by the removal of extracellular calcium. Likewise, in the mouse adrenal medulla slices, orexin A also induced catecholamine release mainly through the activation of OX1R. These results gain insight into our understanding of the pharmacological relevance of orexin system in modulating neuroendocrine functions. PMID- 18395342 TI - Inhibition of MPEP on the development of morphine antinociceptive tolerance and the biosynthesis of neuronal nitric oxide synthase in rat spinal cord. AB - We evaluated the ability of spinally administered 2-methyl-6-(phenylethynyl) pyridine (MPEP), a selective antagonist of the metabotropic glutamate receptor subtype 5 (mGluR5), and 2-chloro-5-hydroxyphenylglycine (CHPG), an mGluR5 agonist, to modulate the antinociceptive action and tolerance of intrathecal (i.t.) morphine infusion in rats, and assessed the expression of spinal nitric oxide synthase (NOS). MPEP co-infused with morphine not only preserved the analgesia and retarded the development of antinociceptive tolerance, but also partially inhibited the up-regulation of spinal nNOS protein. However, the loss of morphine antinociceptive effect and tolerance were accelerated when CHPG and morphine were co-infused, while spinal nNOS activity was significantly up regulated. We hypothesize that activation of mGluR5 and NMDA receptors occurs after the appearance of antinociceptive tolerance to morphine. The activation of these receptors might stimulate an increased concentration of intracellular calcium and activation of PKC, which both play a vital role in the development of morphine antinociceptive tolerance and expression of spinal NOS. The synergistic effect which seems to exist between mGluRs and iGluRs may also contribute to this phenomenon. PMID- 18395343 TI - Onset of feeding at birth--perinatal development of the hypothalamic mechanisms that induce appetite and feeding in the newborn. AB - Orexin-A and -B are hypothalamic peptides which, in the adult brain, are associated with arousal, increased vigilance, and the seeking and ingestion of food. Because the fetus is mostly asleep, and hunger is a physiological state unlikely to arise until birth, we hypothesized that orexigenic neurons in the lateral and dorso-medial hypothalamic areas (LHA, DMH) and their projections to the locus coeruleus (LC) would develop only near the time of birth. We therefore determined orexin expression in fetal sheep, where birth occurs over a tightly regulated interval of 146-148 days gestation. Immunohistochemistry was used to determine the presence and distribution of orexin-A positive fibres and cells at the level of the hypothalamus and LC in fetal (125-137 and 145+ days gestation age) and newborn sheep brains. Orexin was measured by radioimmunoassay in plasma samples taken from chronically catheterised fetal and newborn sheep, and in CSF taken from fetuses and lambs at postmortem. Orexin-A positive cells bodies were observed in the hypothalamus, and orexin-A fibres were found throughout all hypothalamic, thalamic, and brain stem regions of all the fetal and newborn brains examined. Orexin-A was present in plasma and CSF at similar concentrations in fetal and newborn sheep. The presence of orexin in hypothalamic neurons and CSF throughout late gestation suggests that orexinergic regulation of hunger, appetite and the sleep/wake cycle is inhibited, by mechanisms yet to be identified, until the time of parturition. PMID- 18395344 TI - Cilostazol promotes survival of axotomized retinal ganglion cells in adult rats. AB - Cilostazol (CLZ), a selective inhibitor of cyclic nucleotide phosphodiesterase 3, has been shown to reduce neuronal cell death after a transient cerebral infarction. The mechanism for this reduction was suggested to be an elevation of intracellular cAMP or an inhibition of tumor necrosis factor alpha. Optic nerve injury leads to retinal ganglion cell (RGC) death possibly from a deprivation of neurotrophic factors and/or the down-regulation of intracellular cAMP. The purpose of this study was to determine if CLZ can rescue RGCs after optic nerve transection by inhibiting cyclic nucleotide phosphodiesterase 3. To examine this, the mean densities of surviving RGCs after optic nerve transection were determined in retinas that received an intravitreal injection of CLZ and in retinas that received vehicle. Our results showed that the density of surviving RGCs in the retina with intravitreal CLZ were significantly higher than that with vehicle injection on day 7. The CLZ was effective in promoting the survival at more than 0.05% concentration. The neuroprotective effects induced by 0.05% CLZ could be observed even 14 days after optic nerve transection. Furthermore, combined application of protein kinase A (PKA) inhibitor, KT5720 (10 microM) and 0.05% CLZ significantly decreased the density of surviving RGCs compared to that with only 0.05% CLZ. Based on these data, we concluded that CLZ enhances the survival of axotomized RGC in vivo, possibly depending on the activation of PKA pathway. PMID- 18395345 TI - Efficacy and safety of duloxetine for treatment of fibromyalgia in patients with or without major depressive disorder: Results from a 6-month, randomized, double blind, placebo-controlled, fixed-dose trial. AB - The primary objectives of this study were to assess the efficacy and safety of duloxetine for reducing pain severity in fibromyalgia patients with or without current major depressive disorder. This was a 6-month, multicenter, randomized, double-blind, placebo-controlled study. In total, 520 patients meeting American College of Rheumatology criteria for fibromyalgia were randomly assigned to duloxetine (20 mg/day, 60 mg/day, or 120 mg/day) or placebo, administered once daily, for 6 months (after 3 months, the duloxetine 20-mg/day group titrated to 60 mg/day). The co-primary outcome measures were the Brief Pain Inventory (BPI) average pain severity score and Patient Global Impressions of Improvement (PGI-I) score. Safety was assessed via treatment-emergent adverse events, and changes in vital sign, laboratory, and ECG measures. Compared with placebo-treated patients, those patients treated with duloxetine 120 mg/day improved significantly more on the co-primary outcome measures at 3 months (change in BPI score [-2.31 vs -1.39, P<0.001] and PGI-I [2.89 vs 3.39, P=0.004]) and at 6 months (change in BPI [-2.26 vs -1.43, P=0.003] and PGI-I [2.93 vs 3.37, P=0.012]). Compared with placebo, treatment with duloxetine 60 mg/day also significantly improved the co-primary measures at 3 months and BPI at 6 months. Duloxetine was efficacious in patients both with and without major depressive disorder. There were no clinically significant differences between treatment groups in changes in vital signs, laboratory measures, or ECG measures. Study results demonstrated that duloxetine at doses of 60 mg/day and 120 mg/day appears to be safe and efficacious in patients with fibromyalgia. PMID- 18395346 TI - Models for experimental infection of dogs fed with tissue from fetuses and neonatal cattle naturally infected with Neospora caninum. AB - Three models were designed to investigate the development and enteroepithelial phase of Neospora caninum in dogs, and to induce oocyst production by the parasite. In the first model, three dogs were fed raw fetal bovine tissue on two occasions. The bovine fetal tissue had been stored at 4 degrees C for 14 days and 16 days, respectively, and had tested positive for neosporosis using histopathology and immunohistochemistry. In the second model, nine dogs from the beginning of the experiment until euthanasia were fed portions of tissue from bovine fetuses that been stored at 4 degrees C for between 1 and 2 days. Three fetuses had tested positive for neosporosis using histopathology and immunohistochemistry. In the third model, three dogs were fed pieces of raw tissue from two neonatal calves that had not received colostrum, and which had tested positive for antibodies against N. caninum. The brains of these calves were positive for neosporosis by histopathology and immunohistochemistry. In all three models, none of the dogs excreted oocysts of N. caninum, developed intestinal parasites or seroconverted. PMID- 18395347 TI - Impact of previous infestation on dynamics of circulating hypodermin C in cattle artificially infested with Hypoderma lineatum (Diptera: Oestridae). AB - Four groups of cattle were artificially infested with 50 first instar Hypoderma lineatum after either a primary natural infestation or in the absence of a primary infestation. In two groups the primary infestation had been terminated by the application of either an organophosphate insecticide or a macrocyclic lactone parasiticide. Circulating hypodermin C and specific antibodies were measured for 40 weeks after the artificial infestation. Stage specific mortality of the larvae was also monitored. Previously uninfested cattle exhibited typical antibody and antigen profiles during the infestations. Antibodies were first detected on Week 7p.i., they rose to maximum values between Weeks 24 and 25, then declined as larvae reached the back. A second peak occurred at Week 32 and antibody remained more less constant thereafter. Previously infested groups exhibited a dramatic anamnestic response by Week 3p.i. with antibody levels peaking at Week 8. A second peak was noted between Weeks 24 and 26p.i. after which antibody levels declined and then remained relatively stable. The dynamics of circulating hypodermin C in the previously infested cattle resembled those in the previously uninfested cattle. Mortality of first instars did not differ among the four groups. Similarly mortality of second and third instars, in the warble, did not differ although there was a tendency for higher mortality in the previously infested, untreated group. These results reinforce previous work demonstrating the development of a significant immune response during the primary infestation that is reflected in the rapid and substantial production of antibodies upon re infestation. It is significant that a challenge model using subcutaneous injection of newly hatched first instars avoids host immune responses in the skin that result in substantial larval mortality. The current data also support the concept that migrating first instars induce significant reduction in host immune response. There is a peak of antibody production as antigen levels peak during first instar migration to the back. As larvae molt to the second instar and antigen production ceases there is a persistence of antibody which suggests release of the immune response from the suppression induced by the first instar secretory antigens. PMID- 18395348 TI - Field efficacy and safety of a combination of moxidectin and imidacloprid for the prevention of feline heartworm (Dirofilaria immitis) infection. AB - Throughout the end of March to beginning of May 2006, 212 owned cats and 608 owned dogs from a heavy endemic area for canine heartworm (HW) disease in northern Italy have been examined to assess HW infection prevalence. Both cats and dogs were clinically examined and blood samples were taken from each animal to be examined for HW antibody (Ab). Ab-positive cats were further examined for circulating microfilariae, HW antigens (Ag) and by echocardiography (ECHO) to assess the presence of adult worms. Dogs were clinically examined and blood samples taken from each animal were examined for circulating microfilariae and for HW Ag. Ten cats (4.7%) were found Ab positive. Of these, 6 cats were Ag positive (2.6%) and in 4 (1.8%) the worms were visualized by ECHO. HW prevalence in dogs was 36% (221/608). One hundred and seventy-six (29%) were both microfilaraemic and Ag positive, 40 (7%) had occult infections (no circulating microfilariae) and 7 (1%) were microfilaraemic but Ag negative. Upon owners' consent, 132 cats (including cats Ab and/or Ag and ECHO positive) were prophylactically treated against HW disease with an imidacloprid/moxidectin spot on combination (10% imidacloprid/1% moxidectin) monthly administered for 6 months. Cats were re-examined for HW infection in November, 1 month after the last drug administration, and in May-June 2007, 7-8 months after the last treatment. All 122 cats found HW negative before treatment, were found negative at the two examinations at the end of study. The 4 cats Ab positive, 2 cats Ab and Ag positive and 1 Ab, Ag and ECHO positive at the beginning of treatment were found negative. Throughout the treatment, transitory hypersalivation and generic signs of annoyance were reported by owners in 6 cats (4.5%). All signs regressed spontaneously. PMID- 18395349 TI - Determining the zoonotic significance of Giardia and Cryptosporidium in Australian dogs and cats. AB - In a recent study of intestinal parasites in dogs and cats in Australia, Giardia was found to be the most prevalent parasite in dogs. The aim of the current study through the use of molecular tools was to determine the zoonotic significance of the Giardia and Cryptosporidium isolates recovered from dogs and cats during the Australian study. Of the isolates successfully amplified all but one of the Giardia from dogs was either Assemblage C and/or D, with one Assemblage A. Of the cat samples amplified all but one were Assemblage F, with one Assemblage D. We hypothesize that the lack of zoonotic Giardia Assemblages recovered is a result of their being a low prevalence of Giardia in the human population. The Cryptosporidium recovered from dogs and cats was determined to be C. canis and C. felis, respectively, a finding which supports growing evidence that Cryptosporidium in companion animals is of limited public health significance to healthy people. PMID- 18395350 TI - "Reconstruction of the face and neck scar contractures using staged transfer of expanded "super-thin flaps"". PMID- 18395351 TI - Phytophotodermatitis: case reports of children presenting with blistering after preparing lime juice. PMID- 18395352 TI - What grabs his attention but not hers? Estrogen correlates with neurophysiological measures of vocal change detection. AB - Prior research revealed sex differences in the processing of unattended changes in speaker prosody. The present study aimed at investigating the role of estrogen in mediating these effects. To this end, the electroencephalogram (EEG) was recorded while participants watched a silent movie with subtitles and passively listened to a syllable sequence that contained occasional changes in speaker prosody. In one block, these changes were neutral, whereas in another block they were emotional. Estrogen values were obtained for each participant and correlated with the mismatch negativity (MMN) amplitude elicited in the EEG. As predicted, female listeners had higher estrogen values than male listeners and showed reduced MMN amplitudes to neutral as compared to emotional change in speaker prosody. Moreover, in both, male and female listeners, MMN amplitudes were negatively correlated with estrogen when the change in speaker prosody was neutral, but not when it was emotional. This suggests that estrogen is associated with reduced distractibility by neutral, but not emotional, events. Emotional events are spared from this reduction in distractibility and more likely to penetrate voluntary attention directed elsewhere. Taken together, the present findings provide evidence for a role of estrogen in human cognition and emotion. PMID- 18395353 TI - Are sexual side effects of prolactin-raising antipsychotics reducible to serum prolactin? AB - OBJECTIVE: To assess the degree to which sexual side effects (SSE) are associated with prolactin-raising antipsychotics, and to what degree such SSE are reducible to serum prolactin levels. METHOD: A large sample (n=264) of patients treated for 6 weeks with prolactin-raising and prolactin-sparing antipsychotics was assessed for changes in sexual performance in terms of libido, arousal and orgasm using the Antipsychotics and Sexual Functioning Questionnaire. For men also erection and ejaculation were evaluated. At 6 weeks, prolactin levels were assessed and analyzed in relation to sexual performance. RESULTS: Men and women reported SSE (libido and orgasm) with about the same frequency. Prolactin-raising medication induced significantly more SSE than prolactin-sparing medication (adjusted OR=3.4, 95% CI: 1.8, 6.5) with 43% of emerging SSE attributable to prolactin raising medication. When adjusted for serum prolactin, the association between prolactin-raising medication and SSE was reduced but remained significant (OR=2.1, 95% CI: 1.0, 4.5); 27% of emerging SSE remained attributable to prolactin-raising medication. For erectile and ejaculatory dysfunction in men, the attributable fraction due to prolactin-raising medication was 32% before, and 11% after adjustment for serum prolactin. CONCLUSIONS: Around 40% of emerging SSE in schizophrenia are attributable to the prolactin-raising properties of antipsychotic medication. Of this attributable fraction, around one-third to two thirds is directly reducible to the effects of serum prolactin. PMID- 18395354 TI - Old biliverdin reductase: links to insulin resistance and may be a novel therapeutic target. AB - Insulin resistance has been assigned a pivotal role in the pathological progression associated with type 2 diabetes and other chronic metabolic diseases. However, the molecular mechanism involved in this progression is still incompletely understood, and there are still no effective approaches to scavenge it. Many biological molecules, such as ROS, IRS-1, PI3K, have been identified involving in the causes of insulin resistance. Restoring these molecules could ameliorate the phenomenon of insulin resistance. BVR was known for a long time solely as an enzyme reducing biliverdin to bilirubin in the heme metabolic pathway. Presently, accumulative research data showed that BVR was a strong antioxidant enzyme, which could scavenge the excess ROS, and the characteristics of kinase activity and binding with p85 could modulate the biological function of IRS-1 and PI3K. We hypothesize that BVR has a significant role in the progression of insulin resistance, and it will be a promising therapeutic target for treating insulin resistance. PMID- 18395355 TI - Hearing loss after radiotherapy for pediatric brain tumors: effect of cochlear dose. AB - PURPOSE: To determine the effect of cochlear dose on sensorineural hearing loss in pediatric patients with brain tumor treated by using conformal radiation therapy (CRT). PATIENTS AND METHODS: We studied 78 pediatric patients (155 ears) with localized brain tumors treated in 1997-2001 who had not received platinum based chemotherapy and were followed up for at least 48 months. They were evaluated prospectively by means of serial pure-tone audiograms (250 Hz-8 kHz) and/or auditory brainstem response before and every 6 months after CRT. RESULTS: Hearing loss occurred in 14% (11 of 78) of patients and 11% (17 of 155) of cochleae, with onset most often at 3-5 years after CRT. The incidence of hearing loss was low for a cochlear mean dose of 30 Gy or less and increased at greater than 40-45 Gy. Risk was greater at high frequencies (6-8 kHz). In children who tested abnormal for hearing, average hearing thresholds increased from a less than 25 decibel (dB) hearing level (HL) at baseline to a mean of 46 +/- 13 (SD) dB HL for high frequencies, 41 +/- 7 dB HL for low frequencies, and 38 +/- 6 dB HL for intermediate frequencies. CONCLUSIONS: Sensorineural hearing loss is a late effect of CRT. In the absence of other factors, including ototoxic chemotherapy, increase in cochlear dose correlates positively with hearing loss in pediatric patients with brain tumor. To minimize the risk of hearing loss for children treated with radiation therapy, a cumulative cochlear dose less than 35 Gy is recommended for patients planned to receive 54-59.4 Gy in 30-33 treatment fractions. PMID- 18395357 TI - Segmental urethral dosimetry and urinary toxicity in patients with no urinary symptoms before permanent prostate brachytherapy. AB - PURPOSE: To determine whether segmental urethral dosimetry is predictive for the degree of urinary morbidity after prostate brachytherapy in patients with no urinary symptoms before prostate brachytherapy. METHODS AND MATERIALS: Between May 2000 and November 2005, 1,107 patients underwent iodine-125 monotherapy with urethral sparing techniques. A total of 166 patients fulfilled the selection criteria: baseline (International Prostate Symptom Score) IPSS < or =5, no androgen deprivation therapy, and prostate ultrasound planning volumes (PUTV) <45 mL. The median follow-up was 44 months. Urinary morbidity was defined by maximum increase in IPSS, time to IPSS resolution, maximum Radiation Therapy Oncology Group (RTOG) score, time to RTOG resolution, and urinary retention. Surrogate deviated urethra was contoured and doses calculated at the base, mid-prostate, apex, and urogenital diaphragm. Univariate and multivariate analysis was used to evaluate urethral and prostate dosimetry, age, PUTV, and number of needles for their association with urinary morbidity. RESULTS: Urethral dose was fairly constant in all urethra segments except prostate base, where the variation in does was large. On multivariate analysis, higher urethral base D50, V100, and larger PUTV were predictive for higher maximum increase in IPSS. Higher urethral base V100 and larger PUTV predicted for prolonged IPSS resolution. Higher urethral base D50 and larger needle number predicted for longer RTOG resolution. Higher urethral base V100 predicted for RTOG > or =2 toxicity. CONCLUSIONS: Radiation dose to the urethral base, larger PUTV, and needle number, predicted for increased urinary toxicity after prostate brachytherapy. Correlation between urinary morbidity and urethral base dosimetry may reflect a large variation in urethral dose observed at the prostate base. PMID- 18395356 TI - Infusional 5-fluorouracil and ZD1839 (Gefitinib-Iressa) in combination with preoperative radiotherapy in patients with locally advanced rectal cancer: a phase I and II Trial (1839IL/0092). AB - PURPOSE: To report the final data of a Phase I and II study (1839IL/0092) on the combination of an anti-epidermal growth factor receptor drug (gefitinib), infusional 5-fluorouracil, and preoperative radiotherapy in locally advanced, resectable rectal cancer. METHODS AND MATERIALS: Patients received 45 Gy in the posterior pelvis plus a boost of 5.4 Gy on the tumor and corresponding mesorectum. Infusional 5-fluorouracil (5-FU) and gefitinib (250 and 500 mg/day) were delivered during all radiotherapy course. An IORT boost of 10 Gy was allowed. The main endpoints of the study were to establish dose-limiting toxicity (DLT) and to evaluate the rate of pathologic response according to the tumor regression grade (TRG) Mandard score. RESULTS: A total of 41 patients were enrolled. The DLT was not reached in the 6 patients enrolled in the dose escalation part of the study. Of the 33 patients in the Phase II, TRG 1 was recorded in 10 patients (30.3%) and TRG 2 in 7 patients (21.2 %); overall 17 of 33 patients (51.5%) had a favorable endpoint. Overall, Grade 3+ toxicity was recorded in 16 patients (41%); these included Grade 3+ gastrointestinal toxicity in 8 patients (20.5%), Grade 3+ skin toxicity in 6 (15.3%), and Grade 3+ genitourinary toxicity in 4 (10.2%). A dose reduction of gefitinib was necessary in 24 patients (61.5%). CONCLUSIONS: Gefitinib can be associated with 5-FU-based preoperative chemoradiation at the dose of 500 mg without any life-threatening toxicity and with a high pCR (30.3%). The relevant rate of Grade 3 gastrointestinal toxicity suggests that 250 mg would be more tolerable dose in a neaoadjuvant approach with radiotherapy and infusional 5-FU. PMID- 18395358 TI - Comparison of 2D radiographic images and 3D cone beam computed tomography for positioning head-and-neck radiotherapy patients. AB - PURPOSE: To assess the positioning accuracy using two-dimensional kilovoltage (2DkV) imaging and three-dimensional cone beam CT (CBCT) in patients with head and neck (H&N) cancer receiving radiation therapy. To assess the benefit of patient-specific headrest. MATERIALS AND METHODS: All 21 patients studied were immobilized using thermoplastic masks with either a patient-specific vacuum bag (11 of 21, IMA) or standard clear plastic (10 of 21, IMB) headrests. Each patient was imaged with a pair of orthogonal 2DkV images in treatment position using onboard imaging before the CBCT procedure. The 2DkV and CBCT images were acquired weekly during the same session. The 2DkV images were reviewed by oncologists and also analyzed by a software tool based on mutual information (MI). RESULTS: Ninety-eight pairs of assessable 2DkV-CBCT alignment sets were obtained. Systematic and random errors were <1.6 mm for both 2DkV and CBCT alignments. When we compared shifts determined by CBCT and 2DkV for the same patient setup, statistically significant correlations were observed in all three major directions. Among all CBCT couch shifts, 4.1% > or = 0.5 cm and 18.7% > or = 0.3 cm, whereas among all 2DkV (MI) shifts, 1.7% > or = 0.5 cm and 11.2% > or = 0.3 cm. Statistically significant difference was found on anteroposterior direction between IMA and IMB with the CBCT alignment only. CONCLUSIONS: The differences between 2D and 3D alignments were mainly caused by the relative flexibility of certain H&N structures and possibly by rotation. Better immobilization of the flexible neck is required to further reduce the setup errors for H&N patients receiving radiotherapy. PMID- 18395359 TI - Definitive radiotherapy for T1-T2 squamous cell carcinoma of pyriform sinus. AB - PURPOSE: To report the long-term results after definitive radiotherapy (RT) for T1-T2 pyriform sinus squamous cell carcinoma. PATIENTS AND METHODS: The data from 123 patients with T1-T2 pyriform sinus squamous cell carcinoma treated with RT with or without neck dissection between November 1964 and June 2003 were analyzed. The median follow-up for all patients was 3.2 years, and the median follow-up for living patients was 10.7 years. RESULTS: The 5-year local control, locoregional control, freedom from distant metastasis, cause-specific survival, and overall survival rate was 85%, 70%, 75%, 61%, and 35%, respectively. The ultimate local control rate, including successful salvage of RT failure, for T1 and T2 cancer patients was 96% and 94%, respectively. The overall local control rate with a functional larynx was 83%. Pretreatment computed tomography tumor volume data were available for 55 patients. The median computed tomography tumor volume was 4.2 cm(3) (range, 0-22.4). Local control was worse for patients with a tumor volume >6.5 cm(3) compared with those with a smaller tumor volume. Of the 123 patients, 16% developed moderate to severe acute (2%), late (9%), or postoperative (5%) complications. CONCLUSIONS: Local control with larynx preservation after definitive RT for T1-T2 pyriform sinus squamous cell carcinoma likely results in local control and survival similar to that after total laryngectomy or larynx-conserving surgery. Two-thirds of our living patients retained a functional larynx. PMID- 18395360 TI - Prospective clinical trial of bladder filling and three-dimensional dosimetry in high-dose-rate vaginal cuff brachytherapy. AB - PURPOSE: To investigate the effect of bladder filling on dosimetry and to determine the best bladder dosimetric parameter for vaginal cuff brachytherapy. METHODS AND MATERIALS: In this prospective clinical trial, a total of 20 women underwent vaginal cylinder high-dose-rate brachytherapy. The bladder was full for Fraction 2 and empty for Fraction 3. Dose-volume histogram and dose-surface histogram values were generated for the bladder, rectum, and urethra. The midline maximal bladder point (MBP) and the midline maximal rectal point were recorded. Paired t tests, Pearson correlations, and regression analyses were performed. RESULTS: The volume and surface area of the irradiated bladder were significantly smaller when the bladder was empty than when full. Of the several dose-volume histogram and dose-surface histogram parameters evaluated, the bladder maximal dose received by 2 cm(3) of tissue, volume of bladder receiving >/=50% of the dose, volume of bladder receiving >/=70% of the dose, and surface area of bladder receiving >/=50% of the dose significantly predicted for the difference between the empty vs. full filling state. The volume of bladder receiving >/=70% of the dose and the maximal dose received by 2 cm(3) of tissue correlated significantly with the MBP. Bladder filling did not alter the volume or surface area of the rectum irradiated. However, an empty bladder did result in the nearest point of bowel being significantly closer to the vaginal cylinder than when the bladder was full. CONCLUSIONS: Patients undergoing vaginal cuff brachytherapy treated with an empty bladder have a lower bladder dose than those treated with a full bladder. The MBP correlated well with the volumetric assessments of bladder dose and provided a noninvasive method for reporting the MBP dose using three dimensional imaging. The MBP can therefore be used as a surrogate for complex dosimetry in the clinic. PMID- 18395361 TI - Unresectable carcinoma of the paranasal sinuses: outcomes and toxicities. AB - PURPOSE: To evaluate long-term outcomes and toxicity in patients with unresectable paranasal sinus carcinoma treated with radiotherapy, with or without chemotherapy. METHODS AND MATERIALS: Between January 1990 and December 2006, 39 patients with unresectable Stage IVB paranasal sinus carcinoma were treated definitively with chemotherapy plus radiotherapy (n = 35, 90%) or with radiotherapy alone (n = 4, 10%). Patients were treated with three-dimensional conformal radiotherapy (n = 18, 46%), intensity-modulated radiotherapy (n = 12, 31%), or conventional radiotherapy (n = 9, 23%) to a median treatment dose of 70 Gy. Most patients received concurrent platinum-based chemotherapy (n = 32, 82%) and/or concomitant boost radiotherapy (n = 29, 74%). RESULTS: With a median follow-up of 90 months, the 5-year local progression-free survival, regional progression-free survival, distant metastasis-free survival, disease-free survival, and overall survival were 21%, 61%, 51%, 14%, and 15%, respectively. Patients primarily experienced local relapse (n = 25, 64%), mostly within the irradiated field (n = 22). Nine patients developed neck relapses; however none of the 4 patients receiving elective neck irradiation had a nodal relapse. In 13 patients acute Grade 3 mucositis developed. Severe late toxicities occurred in 2 patients with radionecrosis and 1 patient with unilateral blindness 7 years after intensity-modulated radiation therapy (77 Gy to the optic nerve). The only significant factor for improved local progression-free survival and overall survival was a biologically equivalent dose of radiation >/=65 Gy. CONCLUSIONS: Treatment outcomes for unresectable paranasal sinus carcinoma are poor, and combined-modality treatment is needed that is both more effective and associated with less morbidity. The addition of elective neck irradiation may improve regional control. PMID- 18395362 TI - Gemcitabine chemotherapy and single-fraction stereotactic body radiotherapy for locally advanced pancreatic cancer. AB - PURPOSE: Fractionated radiotherapy and chemotherapy for locally advanced pancreatic cancer achieves only modest local control. This prospective trial evaluated the efficacy of a single fraction of 25 Gy stereotactic body radiotherapy (SBRT) delivered between Cycle 1 and 2 of gemcitabine chemotherapy. METHODS AND MATERIALS: A total of 16 patients with locally advanced, nonmetastatic, pancreatic adenocarcinoma received gemcitabine with SBRT delivered 2 weeks after completion of the first cycle. Gemcitabine was resumed 2 weeks after SBRT and was continued until progression or dose-limiting toxicity. The gross tumor volume, with a 2-3-mm margin, was treated in a single 25-Gy fraction by Cyberknife. Patients were evaluated at 4-6 weeks, 10-12 weeks, and every 3 months after SBRT. RESULTS: All 16 patients completed SBRT. A median of four cycles (range one to nine) of chemotherapy was delivered. Three patients (19%) developed local disease progression at 14, 16, and 21 months after SBRT. The median survival was 11.4 months, with 50% of patients alive at 1 year. Patients with normal carbohydrate antigen (CA)19-9 levels either at diagnosis or after Cyberknife SBRT had longer survival (p <0.01). Acute gastrointestinal toxicity was mild, with 2 cases of Grade 2 (13%) and 1 of Grade 3 (6%) toxicity. Late gastrointestinal toxicity was more common, with five ulcers (Grade 2), one duodenal stenosis (Grade 3), and one duodenal perforation (Grade 4). A trend toward increased duodenal volumes radiated was observed in those experiencing late effects (p = 0.13). CONCLUSION: SBRT with gemcitabine resulted in comparable survival to conventional chemoradiotherapy and good local control. However, the rate of duodenal ulcer development was significant. PMID- 18395363 TI - IFNgamma and antibody responses among French nurses during a tuberculosis contact tracing investigation. AB - STUDY: A comparative study which compared PPD skin testing inserted according to the French Society of Pneumology's recommendations and interferon gamma release assay (IGRA) (QuantiFERON((R)) TB Gold In-tube, QF-TB-IT, Cellestis, Carnegie, Australia) was performed during a tuberculosis contact investigation in our hospital. PATIENTS: Nineteen French health-care workers (HCWs) volunteered to participate. All of the HCW enrolled were BCG vaccinated and had a normal chest X ray at entry. RESULTS: Among the HCW, 68.4% were TST positive. By comparison, only 31.6% had a positive QF-TB-IT result. We took advantage of the negative tube and the corresponding plasma for antibody detection by ELISA. None were ELISA positive. Fourteen HCWs were followed up. None of the HCWs accepted a course of antiTB chemoprophylaxis. Despite the difficulty in establishing a trend in kinetics, we saw the complexity of interpretation of a dynamic T-cell response after contact with an index case. CONCLUSION: This initial and first French picture provides us with the observation that only 44% of TST-positive HCW were IGRA positive, and the IGRA test allowed the detection of LTBI in two TST negative HCWs. PMID- 18395364 TI - Intolerance amongst deer species at feeding: roe deer are uneasy banqueters. AB - The function of inter-specific interactions in large herbivore assemblies is under debate. We have studied inter-specific interactions between roe and fallow deer on pastures, to assess whether competition between these Cervidae occurs through behavioural interference. Roe were displaced by fallow deer (i.e. the former moved away from the latter at a distance of >50 m) in 83% of cases (N=127). Ninety-four percentage (N=83) of displacement events occurred while roe deer were feeding; in 50% of these cases (N=78), roe stopped grazing and left the feeding ground. Even when fallow deer did not show any sign of direct aggression to roe, these moved away from fallow in 72% of cases (N=127). Vigilance rate was significantly greater in roe than in fallow deer, irrespectively of the presence of the other species. When roe and fallow deer grazed within 50 m from each other, vigilance rate increased significantly in roe, but not in fallow deer. Roe deer, in a group, were significantly more tolerant of the presence of fallow deer (even in group), than when solitary. Fallow deer seemed to be able to exclude roe deer from feeding sites through behavioural intolerance. PMID- 18395365 TI - Higher magnitude cash payments improve research follow-up rates without increasing drug use or perceived coercion. AB - In a prior study [Festinger, D.S., Marlowe, D.B., Croft, J.R., Dugosh, K.L., Mastro, N.K., Lee, P.A., DeMatteo, D.S., Patapis, N.S., 2005. Do research payments precipitate drug use or coerce participation? Drug Alcohol Depend. 78 (3) 275-281] we found that neither the mode (cash vs. gift card) nor magnitude ($10, $40, or $70) of research follow-up payments increased rates of new drug use or perceptions of coercion. However, higher payments and payments in cash were associated with better follow-up attendance, reduced tracking efforts, and improved participant satisfaction with the study. The present study extended those findings to higher payment magnitudes. Participants from an urban outpatient substance abuse treatment program were randomly assigned to receive $70, $100, $130, or $160 in either cash or a gift card for completing a follow-up assessment at 6 months post-admission (n congruent with 50 per cell). Apart from the payment incentives, all participants received a standardized, minimal platform of follow-up efforts. Findings revealed that neither the magnitude nor mode of payment had a significant effect on new drug use or perceived coercion. Consistent with our previous findings, higher payments and cash payments resulted in significantly higher follow-up rates and fewer tracking calls. In addition participants receiving cash vs. gift cards were more likely to use their payments for essential, non-luxury purchases. Follow-up rates for participants receiving cash payments of $100, $130, and $160 approached or exceeded the FDA required minimum of 70% for studies to be considered in evaluations of new medications. This suggests that the use of higher magnitude payments and cash payments may be effective strategies for obtaining more representative follow-up samples without increasing new drug use or perceptions of coercion. PMID- 18395366 TI - Coordinated expression of clustered cancer/testis genes encoded in a large inverted repeat DNA structure. AB - Cancer/testis antigens (CTA) are expressed in cancers and testis or placenta only and, therefore are considered promising targets for cancer immunotherapy and diagnosis. One family of CTA is the MAGEA family which comprises 13 members and was shown to be expressed synchronously with members from the CSAG (TRAG-3) family of CTA. The MAGEA genes are arranged in 4 subclusters located on the X chromosome. Subcluster III exposes a remarkable gene organization with an inverted repeat (IR) DNA structure of a triplicated couplet of a MAGEA gene and a CSAG gene. Analyzing the mRNA expression pattern of all genes of the MAGEA and CSAG family of cancer/testis genes, we show that the MAGEA and CSAG genes encoded in the large IR are expressed coordinately and independent from the MAGEAs encoded outside the IR. These results reinforce our hypothesis that the large MAGEA/CSAG-IR DNA structure has an impact on the regulation of gene expression. PMID- 18395367 TI - Functional diversification of a protease inhibitor gene in the genus Drosophila and its molecular basis. AB - The mutually exclusive use of alternative reactive site loop (RSL) cassettes due to alternative splicing of serpin (serine protease inhibitor) gene transcripts is a widespread strategy to create target-selective protease inhibitors in the animal kingdom. Since molecular basis and evolution of serpin RSL cassette exon amplification and diversification are unexplored, the exon-intron organization of the serpin gene spn4 from 12 species of the genus Drosophila was studied. The analysis of the gene structures shows that both number and target enzyme specificities of Spn4 RSL cassettes are highly variable in fruit flies and includes inhibitor variants with novel antiproteolytic activities in some species, indicating that RSL diversity is the result of adaptive evolution. Comparative genomics suggests that interallelic gene conversion and/or recombination events contribute to RSL cassette exon amplification. Due to an intron that is located at the most suitable position within the RSL region, multiple inhibitors can be formed in an economic manner that are both efficient and target-selective, allowing fruit flies to control an astonishing variety of proteases with different cleavage chemistry and evolutionary ancestry. PMID- 18395368 TI - Molecular evolution and characterization of hepcidin gene products in vertebrates. AB - Antimicrobial peptides (AMPs) include a diverse group of gene-encoded molecules that play a role in innate defense in many organisms. Evolutionary analyses of the AMP genes can be challenging because of gene duplication and diversification. Recently discovered, hepcidins are small, cysteine-rich antimicrobial peptides that also function as hormonal regulators of iron homeostasis. In this paper we investigated the organization, expression and molecular evolution of hepcidin. From searches of the literature and public genomic databases we collected 68 different hepcidin gene products from 51 different species, all among the vertebrates. Although some species have multiple hepcidin homologues, we suggest that each contains only one copy that functions as an iron regulator. Despite the recent report of hepcidin sequences in the pigeon (Fu, Y.M., Li, S.P., Wu, Y.F., Chang, Y.Z., 2007. Identification and expression analysis of hepcidin-like cDNAs from pigeon (Columba livia). Mol. Cell. Biochem. 305, 191-197.), searches of the chicken genomic, EST, and HTGS databases did not reveal any evidence of the presence of this gene in birds. This, along with the absence of reported avian transferrin receptor 2 and hemojuvelin sequences, suggests that iron homeostasis in birds may be regulated by an alternative mechanism. PMID- 18395369 TI - Antimicrobial resistance and genetic characterization of fluoroquinolone resistance of Pseudomonas aeruginosa isolated from canine infections. AB - Infections with antimicrobial-resistant bacteria are a great challenge in both human and veterinary medicine. The purpose of this study was to determine antimicrobial susceptibility of 106 strains of Pseudomonas aeruginosa isolated from dogs with otitis and pyoderma from 2003 to 2006 in the United States. Three antimicrobial panels, including 6 classes and 32 antimicrobial agents, were used. A wide range of susceptibility patterns were noted with some isolates being resistant to between 8 and 28 (mean 16) of the antimicrobials tested. Among the beta-lactams, all isolates were resistant to ampicillin, cefoxitin, cefpodoxime, cephalothin and cefazolin followed by amoxicillin/clavulanic acid (99%), ceftiofur (97%), ceftriaxone (39%), cefotaxime (26%), and cefotaxime/clavulanic acid (20%), whereas less than 7% of isolates were resistant to ceftazidime/clavulanic acid, ceftazidime, piperacillin/tazobactam or cefepime. Two isolates were resistant to the carbapenems. Among the quinolones and fluoroquinolones, the most isolates were resistant to naladixic acid (96%), followed by orbifloxacin (52%), difloxacin (43%), enrofloxacin (31%), marbofloxacin (27%), gatifloxacin (23%), levofloxacin (21%), and ciprofloxacin (16%). Among the aminoglycosides, the most resistance was seen to kanamycin (90%), followed by streptomycin (69%), gentamicin (7%), and amikacin (3%). Of the remaining antimicrobials 100% of the isolates were resistant to chloramphenicol followed by tetracycline (98%), trimethoprim/sulfamethoxazole (57%), and sulfisoxazole (51%). Point mutations were present in gyrA, gyrB, parC, and/or parE genes among 34 of the 102 naladixic acid-resistant isolates. Two isolates contained class 1 integrons carrying aadA gene conferring streptomycin and spectinomycin resistance. The findings suggest that many antimicrobial agents commonly used in companion animals may not constitute appropriate therapy for canine pseudomonas infections. PMID- 18395370 TI - Excretion of BSE and scrapie prions in stools from murine models. AB - Faeces from infected animals have been suggested as a potential source of contamination and transmission of prion diseases in the environment. This work describes the development of a procedure for the detection of PrP(res) in stools which is based on a detergent-based extraction and immunoprecipitation (IP). The procedure was evaluated by analyzing TSE-spiked sheep and mice faeces, and proved to be specific for PrP(res) with sensitivities of 5-10 microg of infected brain tissue. In order to analyze the shedding of prions, we studied stools from orally inoculated mice over 4-days post-inoculation and also stools from terminally sick scrapie-infected mice. PrP(res) was only detected in stools shortly after the oral ingestion of TSE agents. The procedure described could be a useful tool for studying the excretion of prions and for evaluating potential environmental contamination by prions. PMID- 18395372 TI - Verminoside- and verbascoside-induced genotoxicity on human lymphocytes: involvement of PARP-1 and p53 proteins. AB - Verminoside and verbascoside are natural compounds present in plants used in traditional medicine. They exhibit several biological activities including anti inflammatory, anti-bacterial and anti-tumor properties. The potential applications of these compounds as ingredients in pharmaceutical formulations and cosmetics prompted us to investigate on cytotoxic and genotoxic activity of verminoside and verbascoside on human lymphocytes using genetic toxicity assays recommended in preclinical studies by the US Food and Drug Administration (FDA). We analyzed chromosome aberrations (CAs) and sister chromatid exchanges (SCEs) as well as the mitotic index (MI) and cell viability after the treatments with verminoside and verbascoside. This report is the first to clearly demonstrate a significant increase of structural CAs and SCEs on normal human lymphocytes associated with a reduction of the MI in both verminoside- and verbascoside treated cells. Moreover, we observed enhanced protein expression levels of PARP-1 and p53 that are key regulatory proteins involved in cell proliferation and DNA repair. Interestingly, mass spectrometric analysis of the compounds in the culture supernatants also showed that verminoside remained unchanged during the culture period while verbascoside was hydrolyzed to its derivative, caffeic acid and the last one seems to be responsible for the observed biological activity. PMID- 18395371 TI - Immunochemical assessment of deoxynivalenol tissue distribution following oral exposure in the mouse. AB - Deoxynivalenol (DON or vomitoxin) is a trichothecene mycotoxin commonly found in cereal grains that adversely affects growth and immune function in experimental animals. A competitive enzyme-linked immunosorbent assay (ELISA) was used to monitor the kinetics of distribution and clearance of DON in tissues of young adult B6C3F1 male mice that were orally administered 25mg/kg bw of the toxin. DON was detectable from 5 min to 24h in plasma, liver, spleen and brain and from 5 min to 8h in heart and kidney. The highest DON plasma concentrations were observed within 5-15 min (12 microg/mL) after dosing. There was rapid clearance following two-compartment kinetics (t(1/2)alpha=20.4 min, t 1/2 beta=11.8h) with 5% and 2% maximum plasma DON concentrations remaining after 8 and 24h, respectively. DON distribution and clearance kinetics in other tissues were similar to that of plasma. At 5 min, DON concentrations in mug/g were 19.5+/-1.9 in liver, 7.6+/-0.5 in kidney, 7.3+/-0.8 in spleen, 6.8+/-0.9 in heart and 0.8+/ 0.1 in the brain. DON recoveries in tissues by ELISA were comparable to a previous study that employed (3)H-DON and 25mg/kg bw DON dose. The ELISA was further applicable to the detection of DON in plasma of mice exposed to the toxin via diet. This approach provides a simple strategy that can be used to answer relevant questions in rodents of how dose, species, age, gender, genetic background and route/duration of exposure impact DON uptake and clearance. PMID- 18395373 TI - Effect of hen's egg yolk on capacitation and acrosome reaction of diluted canine spermatozoa. AB - The aim of this study was to investigate the influence of progesterone, cholesterol and calcium (Ca(2+)) in an egg-yolk-containing extender on capacitation and acrosome reactions (AR) of diluted canine spermatozoa during 4 days of cooled-storage. For this purpose, we first investigated the effect of supplementation of a Tris-citrate-fructose buffer (TCF) with progesterone in a final concentration of 0.1, 0.2 and 1.0 microg progesterone/ml TCF-diluted semen. We then compared the effects of TCF and the same buffer-containing 20% egg yolk (TCF-EY). In egg yolks and the TCF-EY, progesterone was measured by enzyme immunoassay, cholesterol by enzymatic colorimetry and Ca(2+) by flame atomic absorption spectrophotometry. For both experiments, ejaculates from eight dogs were used. For the comparison of diluents, one ejaculate was divided and one half diluted with TCF, the other with TCF-EY. One half of each TCF- and TCF-EY-diluted sample was evaluated immediately (D1), the other after storage for 4 days at +4 degrees C (D4). In diluted semen, motility and viability were measured by a computer assisted sperm analyzer (CASA; Sperm Vision, Minitub, Germany), capacitation and AR were evaluated with a modified chlortetracycline assay (CTC) and the AR additionally by flow cytometry. RESULTS: Supplementation of progesterone revealed, that between D1 and D4, total and progressive motility decreased with all progesterone concentrations, while viability as well as percentage of capacitated and acrosome reacted spermatozoa stayed constant. Progesterone-, cholesterol- and Ca(2+) concentrations in egg yolks were 524.8+/ 131.4 ng/g, 13.9+/-2.03 mg/g and 1.27+/-0.17 mg/g, respectively. In the TCF-EY diluent, the respective values were 210.9 ng/g, 2.52 mg/g and 1.1mg/g. In TCF semen, at D1, motility and viability were significantly higher than in TCF-EY samples (p<0.05), however at D4, no significant differences were detectable. Further, in TCF-semen, percentages of spermatozoa with intact membranes decreased significantly (p<0.05) and capacitated spermatozoa increased (p<0.05), which was not seen in TCF-EY-samples. In all samples, low percentages of AR were detected and after 4 days, the highest value of AR in TCF-EY-samples was 5.3% on average, as detected by flow cytometry. We therefore conclude that progesterone from egg yolk in routine extenders does not substantially influence semen longevity or AR of canine semen during cold-storage for 4 days. In contrary, egg yolk seems to prevent a significant increase in capacitated spermatozoa. PMID- 18395374 TI - Role of a novel multifunctional excipient poly(ethylene glycol)-block-oligo(vinyl sulfadimethoxine) in controlled release of lysozyme from PLGA microspheres. AB - This study investigated the effect of ion-pairing of anionic polyelectrolytes: our novel poly(ethylene glycol)-block-oligo(vinyl sulfadimethoxine) (PEG-OVSDM) and poly(ethylene glycol)-block-poly(l-aspartic acid) (PEG-PAA) with cationic lysozyme on retention of protein stability during emulsification. Soluble lysozyme recovery after exposure to the deleterious interface was 42-88% (when ion-paired with PEG-OVSDM, PEG-OVSDM concentration dependent) compared to only 30% for free lysozyme. PEG-OVSDM provided a higher stabilization of lysozyme than PEG-PAA (36-60%). Lysozyme when recovered in the aqueous phase and analyzed by chromatography, enzymatic assay, fluorescence, and mass spectrometry showed no significant physicochemical change when compared with a lysozyme standard. Lysozyme was incorporated into poly(lactide-co-glycolide) (PLGA) microspheres via the typical double emulsion method. Incorporation of lysozyme complexes led to a higher encapsulation efficiency and loading amount, and a lower incidence of insoluble lysozyme aggregates compared to the control microspheres containing lysozyme only. More significantly, ion-pairing was able to dramatically reduce the initial lysozyme release to 18% compared with 50% from control microspheres and provided an overall better control of protein release. PEG-PAA was less effective than PEG-OVSDM in controlling the release probably due to weaker interactions between this polyelectrolyte and lysozyme. Manipulation of such polyelectrolyte-protein complexation may play a role in protein-controlled delivery. PMID- 18395375 TI - [Efavirenz versus indinavir among HIV-1 naive patients in Abidjan (Ivory Coast)]. AB - OBJECTIVE: The authors had for aim to compare the therapeutic efficiency and tolerance of 2 NRTI+efavirenz (EFV) versus 2 NRTI+indinavir (IDV) in HIV infected adults in Abidjan. METHODS: A retrospective and multicentric study was made on 327 HIV-1 naive patients, 142 in the EFV group and 185 in the IDV group followed in Abidjan from November 1998 to December 2003. The analysis concerned clinical advents (opportunistic infections) and immunovirological parameters (CD4, viral load). Patients received 2 NRTI such as AZT+3TC or D4T+3TC combined either with EFV or IDV. The principal judgement criterion was therapeutic failure. We assessed the percentage of patients with undetectable viral load and the frequency of grade 3-4 adverse effects after 24 months of follow-up. RESULTS: Clinical improvement of patients' state and regression of opportunistic infections were identical in the two groups. The average gain of CD4 was superior to 177 in EFV versus +219 in IDV (p=0.004). The percentage of patients with undetectable viral load was 66% for EFV versus 59% for IDV (p=0.04). The frequency of adverse effects was more elevated with EFV than IDV, 39% versus 23% (p=0.002) initially, but seemed to decrease later. CONCLUSION: HAART with EFV is at least as efficient as with IDV in terms of reduction of viral load and increased CD4 count and is an excellent low-cost first line treatment. PMID- 18395376 TI - [Epidemiological aspects of human Q fever in Indre-et-Loire between 2003 and 2005 and comparison with caprine Q fever]. AB - OBJECTIVE: Chronic infection is the major risk of Q fever. C. burnetii infections result from the inhalation of contaminated aerosols. Indre-et-Loire is a rural French area with numerous goat farms. We evaluated human Q fever epidemiology and compared it with Q fever in goats. DESIGN: This retrospective study was made between 2003 and 2005. The diagnosis of C. burnetii infection was based on serologic findings from all the subdivision laboratories. Antibodies were detected by using indirect immunofluorescence. Farm animal data was processed by ELISA on blood samples from goats and cattle after Q fever related abortion in 2006 and results of PCR-processed milk samples from 156 goat farms. RESULTS: Forty human cases were studied: 38 acute Q fever (11 pneumonia, 10 hepatitis, 10 pneumonia with hepatitis, two isolated fever) and six chronic Q fever (four endocarditis). Sixteen patients (40%) had been professionally exposed, 10 (25%) of whom were goat farmers. Eight (20%) had been in contact with placenta. All the human cases were located in the south of Indre-et-Loire. Twenty percent of the volunteer goat farms had at least one milk sample positive for Q fever by PCR. Forty-nine of the 75 goat abortion samples were positive in ELISA. Ninety-two of the goat farms with positive samples were located in the south of Indre-et-Loire. CONCLUSION: This study revealed similar location of human and caprine Q fever. Identifying such geographical correlation may lead to improving prevention and detection. PMID- 18395377 TI - [Bacteremia caused by Kocuria kristinae in a patient with acute leukaemia]. PMID- 18395378 TI - [Frequency and antibiotic susceptibility of bacteria identified in urine]. AB - OBJECTIVE: The authors had for aim to evaluate the frequency of isolation and the antibiotic susceptibility of bacteria responsible for urinary infections among in and out-patients. METHODS: A retrospective study was carried out at the Microbiology Laboratory of the Mohammed-V Military Instruction Hospital in Rabat, on bacteria isolated from 896 positive cytobacteriologic examinations of urine, from January 1 to December 31, 2005. RESULTS: Among the 7472 urinary samples, 896 answered the criteria of urinary infection (12%). Urinary infections were more frequent in in-patients (57.8%; sex-ratio F/M=1:16). Enterobacteria were the most frequent (85%) and especially Escherichia coli (44.7%). Gram positive bacteria (11.6%) were more frequently Staphylococcus saprophyticus (3.1%). E. coli was highly resistant to amoxicillin, and sulfamethoxazole, whereas quinolones and aminosides maintained a good activity and third generation cephalosporines remain active (97% of susceptibility). Among the enterobacteria present, 3.2% were resistant to third generation cephalosporines with extended spectrum betalactamase. PMID- 18395379 TI - [Molecular strain typing contribution to epidemiology of tuberculosis in Limousin (1998 to 2006)]. AB - OBJECTIVES: We conducted a molecular epidemiology of Mycobacterium tuberculosis in Limousin, a French area with a low incidence of tuberculosis (4.8/100,000 inhabitants in 2005) to define the molecular diversity and the pattern of transmission. DESIGN: Two hundred and fifty-nine strains were isolated (each strain corresponds to one patient) from 1998 to 2006. Both spoligotyping and MIRU15 were chosen for our study because of their discriminatory power. RESULTS: Only 165 medical records were available: 99M/66F, mean age 56.4 years (14-94), 32.7% foreign-born patients, 16.9% homeless or living in shelters, 21.8% of immunocompromised patients (three HIV positive), 14.5% of alcohol addicts. Pulmonary manifestations were predominant (81.8%) with 45.1% of positive smears. Two strains among the 259 presented a multidrug resistance. Spoligotyping identified 136/259 spoligotypes (110 unique, 26 clusters composed of two to 36 isolates); within these 26 clusters, ST53 (n=36) and ST50 (n=19) were the most frequent. Three major families were observed as follow: T1 (30%), Haarlem (30%) and LAM (20%). MIRU15 identified 28/36 isolates in the ST53 group and 14/19 in the ST50 group. Eleven clusters (32 strains) with identical ST-MIRU15 were obtained with a proved case of recent transmission. Alcohol dependence, immunosuppression and pulmonary infections seem to be involved in transmission factors. CONCLUSION: M. tuberculosis strains isolated in Limousin are characterized by their high genetic diversity. The rate of recent transmission (8.1%) is low and therefore a reactivation process is predominant in this area. PMID- 18395380 TI - Antibiotic susceptibility and mechanisms of beta-lactam resistance among clinical strains of Pseudomonas aeruginosa: first report in Algeria. AB - OBJECTIVE: Pseudomonas aeruginosa is a major causative agent of hospital infections. Studies on this subject being rare in Algeria, we determined the antibiotic susceptibility of P. aeruginosa and investigated the mechanisms of beta-lactam resistance and the spread of multidrug resistant strains in the university affiliated Hospital of Tlemcen (Algeria). DESIGN: One hundred and ninety-nine consecutive strains of P. aeruginosa were collected between November 2005 and February 2007. MICs of antibiotics were measured by the agar dilution method. The resistance mechanisms to beta-lactams were identified phenotypically or by molecular methods (isoelectrofocusing, PCR and sequencing). Strains expressing a secondary beta-lactamase were serotyped and genotyped (Random Amplified Polymorphic DNA). RESULTS: The proportion of susceptible isolates were: ticarcillin (56%), piperacillin-tazobactam (81%), ceftazidime (88%), cefepime (80%), aztreonam (64%), imipenem (65%), amikacin (83%), tobramycin (81%) and ciprofloxacin (97%) according to the French CASFM breakpoints. Resistance to beta lactams was linked to the production of transferable beta-lactamases (16%), overproduction of cephalosporinase AmpC (12%) and/or non-enzymatic mechanisms such as the loss of porin OprD (35%) and overproduction of the active efflux system MexAB-OprM (24%). High level resistance to ticarcillin was due to the expression of beta- lactamase OXA-10 alone or associated with TEM-110. A genotypic analysis revealed the spread of a multidrug resistant epidemic clone expressing these two acquired beta-lactamases in the surgical ICU. CONCLUSIONS: This study shows that resistance to antibiotics, in particular to imipenem of P. aeruginosa, is becoming a cause of concern in the Hospital of Tlemcen. PMID- 18395381 TI - [New use of primaquine for malaria]. AB - Primaquine is the only available drug to treat Plasmodium vivax liver stages (hypnozoites). It has been used for more than five decades and is now included in an increasing number of clinical guidelines. The major concern is induced hemolysis when administered to glucose-6-phosphate-dehydrogenase deficient patients. Primaquine could be used for causal prophylaxis during and after exposure or for presumptive antirelapse therapy (PART) in case of high exposure to P. vivax. A radical cure is used to avoid relapse for patients with a confirmed bloodstream infection with P. vivax or P. ovale. In France, primaquine is not approved for prevention and treatment and its use requires a specific temporary authorization. PMID- 18395382 TI - [Early onset neonatal infections in the South of the Reunion Island: Incidence and use of 2002 ANAES risk criteria]. AB - OBJECTIVES: The aim of this study was to describe the incidence of early onset neonatal infections (EONI) in the southern part of the Reunion Island, and to study the application of ANAES criteria. PATIENTS AND METHODS: A cross-sectional study was made of data collected for all live births having occurred between 1st January 2001 and 31st December 2004. RESULTS: Four hundred and thirty-seven in 16,071 neonates (out of 21,231 live births) presented with a certain or probable EONI, accounting for a regional rate of 20 per thousand (CI95 % 18-23 per thousand). Among 437 EONIs, group B streptococcus (GBS) was reported in 70.5% of the cases (n=308), Gram negative bacteria in 19.9% (n=87), of which nearly two thirds of Escherichia coli (n=56). Applying ANAES criteria led to identify 380 EONIs among 437 proven infections (sensitivity: 87%, specificity: 26%). A logistic regression analysis identified eight EONI predictors for the 7015 neonates for whom the mother GBS screening was documented: GBS positive vaginal culture (OR 4.2; CI95% 3.3-5.4), unexplained preterm birth less than 35 weeks (OR 5.7; CI95% 3.7-8.7), prolonged rupture of membranes greater than or equal to 18 hours (OR 2.1; CI95% 1.4-3.0), maternal fever greater than or equal to 37.8 degrees C (OR 3.2; CI95% 2.3-4.5), fetal tachycardia greater than or equal to 160 ppm (OR 2.7; CI95% 1.8-4.0), and thin (OR 1.6; CI95% 1.2-2.1) or thick meconium stained amniotic fluid (OR 3.0; CI95% 2.1-4.5) or fetid fluid (OR 14.8; CI95% 4.2 51.8). CONCLUSION: The incidence of EONIS far exceeded that observed in metropolitan France, and the ANAES criteria lack sensitivity and specificity. PMID- 18395383 TI - [Demographic, clinical, immunovirological and therapeutic features of 8714 HIV infected French patients included in the Nadis Hospital Cohorte in 2006]. AB - OBJECTIVES: The authors had for aim to describe demographic, immunovirilogical and therapeutic characteristics of HIV infected patients enrolled in a French clinical cohort. STUDY DESIGN: A cross-sectional analysis was performed on 30 September 2006, among patients followed in seven French University Medical Centers using the Nadis computerized medical file. RESULTS: Among 8714 patients enrolled (median age 43 years: 15-86, sex ratio 2.37), sexual transmission was the most frequent route of infection (heterosexual: 39.3%, homosexual: 34.8%). HIV and hepatitis C (HCV) or B (HBV) co-infection rates were at 19.2 and 5.8%, respectively. The number of patients who had a triple infection with HIV-HBV-HCV were 1.7%. CDC aids classification was A: 56.7%, B: 19.6%, C: 23.7%. We observed a higher proportion of female patients and an increase of the median age. The number of patients receiving antiretroviral therapy (ARV) at the study date were 81.7%, 11.7% were ARV-naive and 6,6% on a treatment interruption. Under ARV, the median CD4 count was 478cells per millimetre cube (1 to 2166) and 84.8% of patients had an undetectable viral load (VL). Among these patients, 10% had a CD4 cell count inferior or equal to 200 per millimetre cube. Patients followed in centers that participated in this study were different regarding sex, transmission routes, and HBV or HCV co-infection rates, but not regarding the proportion of patients with undetectable VL. ARV combination included more frequently protease inhibitors than nonnucleosidic reverse transcriptase (50% versus 26.2%). CONCLUSION: Among the 8714 HIV-infected patients on ARV, 85% had a VL inferior or equal to 400 per millilitre, 10% of whom had CD4 cell counts inferior or equal to 200 per millimetre cube. The proportion of patients on ARV with undetectable VL was comparable in centers who participated in this study. PMID- 18395384 TI - [Clostridium clostridiiforme vertebral osteomyelitis]. PMID- 18395386 TI - Endovascular reconstruction of completely occluded left brachiocephalic vein in haemodialysis patients--tips and tricks. AB - Venous stenoses or occlusions are common clinical problems in haemodialysis patients and interventional endovascular techniques are highly effective and offer minimally invasive solutions. Not only venous stenoses but also complete occlusions can be treated by endovascular techniques. Tips and tricks for recanalisation in these difficult topographic locations are presented. PMID- 18395385 TI - Increased iron content and RNA oxidative damage in skeletal muscle with aging and disuse atrophy. AB - Muscle atrophy with aging or disuse is associated with deregulated iron homeostasis and increased oxidative stress likely inflicting damage to nucleic acids. Therefore, we investigated RNA and DNA oxidation, and iron homeostasis in gastrocnemius muscles. Disuse atrophy was induced in 6- and 32-month old male Fischer 344/Brown Norway rats by 14 days of hind limb suspension (HS). We show that RNA, but not DNA, oxidative damage increased 85% with age and 36% with HS in aged muscle. Additionally, non-heme iron levels increased 233% with aging and 83% with HS at old age, while staining for free iron was strongest in the smallest fibers. Simultaneously, the mRNA abundance of transferrin receptor-1 decreased by 80% with age and 48% with HS for young animals, while that of the hepcidin regulator hemojuvelin decreased 37% with age, but increased about 44% with disuse, indicating a dysregulation of iron homeostasis favoring increased intracellular free iron in atrophied muscles. RNA and DNA concentrations increased with age and were negatively correlated with muscle mass, whereas protein concentrations decreased with aging, indicating a preferential loss of protein compared to nucleic acids. Furthermore, xanthine oxidase activity increased with age, but not with HS, while mRNA abundance of the Y box-binding protein-1, which has been suggested to bind oxidized RNA, did not change with age or HS. These results suggest that RNA oxidation, possibly mediated by increased non-heme iron, might contribute to muscle atrophy due to disuse particularly in aged muscle. PMID- 18395387 TI - Breast cancer with low FDG uptake: characterization by means of dual-time point FDG-PET/CT. AB - BACKGROUND: Malignant breast lesions usually are differentiated by FDG-PET with a semiquantitative FDG standardized uptake value (SUV) of 2.5. However, the frequency of breast cancer with an SUV of less than or equal to 2.5 is noteworthy, and often present diagnostic challenges. This study was undertaken to evaluate the accuracy of dual-time point FDG-PET/CT with FDG standardized uptake value (SUV) calculation in the characterization of such breast tumors. METHODS: Forty-nine female patients with newly diagnosed breast cancer were found to have primary breast cancer with minimally increased FDG uptake and met the criteria for inclusion in this study by having borderline levels of increased FDG uptake (SUV max less than or equal to 2.5) in the initial FDG-PET/CT images. Consequently, they underwent further delayed phase FDG-PET/CT scan for better evaluation of the disease. RESULTS: Of the 49 cancer lesions; the majority were found to have rising or unvarying dual-time changes in SUV max (75.5%). The median value of SUV max increases by 25% between the early and delayed scan. The means+/-S.D. of the SUV max1, the SUV max2, and the Delta SUV max% were 1.2+/ 0.6%, 1.3+/-0.9%, and 5.1+/-22.4%, respectively. The receiver-operating characteristic (ROC) analysis proved that the highest accuracy for characterization of malignant breast lesions was obtained when a Delta SUV max% cut-off value 0.0% was used as criteria for malignant FDG uptake-change over time with sensitivity 75.5%, and false-positive rate 20.4%. CONCLUSION: These results suggested that dual-time FDG-PET/CT imaging with standardized uptake value (SUV) estimation can improve the accuracy of the test in the evaluation of breast cancer with low FDG uptake. PMID- 18395388 TI - Evaluation of carotid vessel wall enhancement with image subtraction after gadobenate dimeglumine-enhanced MR angiography. AB - OBJECTIVES: This study was aimed at testing the value of image subtraction for evaluating carotid vessel wall enhancement in contrast-enhanced MR angiography (MRA). MATERIALS AND METHODS: IRB approval was obtained. The scans of 81 consecutive patients who underwent carotid MRA with 0.1 mmol/kg of gadobenate dimeglumine were reviewed. Axial carotid 3D T1-weighted fast low-angle shot sequence before and 3 min after contrast injection were acquired and subtracted (enhanced minus unenhanced). Vessel wall enhancement was assigned a four-point score using native or subtracted images from 0 (no enhancement) to 3 (strong enhancement). Stenosis degree was graded according to NASCET. RESULTS: With native images, vessel wall enhancement was detected in 20/81 patients (25%) and in 20/161 carotids (12%), and scored 2.0+/-0.6 (mean+/-standard deviation); with subtracted images, in 21/81 (26%) and 22/161 (14%), and scored 2.5+/-0.6, respectively (P<0.001, Sign test). The overall stenosis degree distribution was: mild, 41/161 (25%); moderate, 77/161 (48%); severe, 43/161 (27%). Carotids with moderate stenosis showed vessel wall enhancement with a frequency (17/77, 22%) significantly higher than that observed in carotids with mild stenosis (1/41, 2%) (P=0.005, Fisher exact test) and higher, even though with borderline significance (P=0.078, Fisher exact test), than that observed in carotids with severe stenosis (4/43, 9%). CONCLUSION: Roughly a quarter of patients undergoing carotid MRA showed vessel wall enhancement. Image subtraction improved vessel wall enhancement conspicuity. Vessel wall enhancement seems to be an event relatively independent from the degree of stenosis. Further studies are warranted to define the relation between vessel wall enhancement and histopathology, inflammatory status, and instability. PMID- 18395389 TI - Guaiazulene in health care products: determination by GC-MS and HPLC-DAD and photostability test. AB - A liquid chromatographic method with UV detection (HPLC-DAD) and a gas chromatographic method coupled with mass spectrometry (GC-MS) have been developed for the determination of guaiazulene (GA) in complex matrices such as creams and toothpastes. A solid phase extraction (SPE) sample pre-treatment on a polymeric sorbent (Strata-X polymer) was applied before the HPLC analyses, which were performed on a XTerra C8 stationary phase, using a mobile phase consisting of acetonitrile-water 50:50 (v/v). For GC-MS analyses, solid-liquid extraction (creams) and SPE (toothpastes) were applied. The proposed methods, based on techniques with different selectivity, were validated and both proved to be suitable to obtain an unambiguous identification and reliable determination of GA in commercial health care products (creams and toothpastes), giving concordant results. Moreover, the described methods can offer a useful analytical support for photostability studies of GA, a photolabile natural compound, in creams. PMID- 18395390 TI - Pharmacokinetics and tissue distribution study of orientin in rat by liquid chromatography. AB - A simple HPLC-UV method was established for the determination of orientin in plasma and different tissues of rat (heart, liver, spleen, lung, kidney, brain, stomach and small intestine). The separation was achieved by HPLC on a C(18) column with a mobile phase composed of acetonitrile-0.1% acetic acid (20:80, v/v), UV detection was used at 348 nm. Good linearity was found between 0.250 50.0 microg/ml (r(2) = 0.9966) for plasma samples and 0.050-50.0 microg/ml (r(2)> or =0.9937) for the tissue samples, respectively. Within- and between-day precisions expressed as the relative standard deviation (R.S.D.) for the method were 2.3-9.6% and 3.0-7.4%, respectively. The relative recoveries of orientin ranged from 95.4 to 100.6% for plasma and 93.1 to 107.9% for tissue homogenates. The developed method was successfully applied to the pharmacokinetics and tissue distribution research after intravenous administration of a 20 mg/kg dose of orientin to healthy Sprague-Dawley rats. The main pharmacokinetics parameters obtained presented that orientin was quickly distributed and eliminated within 90 min after intravenous administration. The tissue distribution results showed that liver, lung and kidney were the major distribution tissues of orientin in rats, and that orientin had difficulty in crossing the blood-brain barrier. It was also found that there was no long-term accumulation of orientin in rat tissues. PMID- 18395391 TI - Microdialysis as a tool to determine free kidney levels of voriconazole in rodents: a model to study the technique feasibility for a moderately lipophilic drug. AB - Microdialysis has been employed for the in vivo measurement of endogenous compounds and a variety of drugs in different tissues. The applicability of this technique can be limited by drug lipophilicity which can impair the diffusion through dialysis membrane. The objective of this study was to evaluate the feasibility of using microdialysis to study kidney penetration of voriconazole, a moderately lipophilic antifungal triazolic agent (LogD7.4=1.8). Microdialysis probe recoveries were investigated in vitro by dialysis and retrodialysis using four different drug concentrations (0.1-2 microg/mL) at five flow rates (1-5 microL/min). Recoveries were dependent on the method used for the determination as well as on the flow rate, but independent of drug concentration. The average apparent recoveries determined by dialysis and retrodialysis, at flow rate of 2 microL/min, were 21.1+/-1.5% and 28.7+/-2.0%, respectively. Recovery by retrodialysis was bigger than the recovery by dialysis. The average apparent dialysis/retrodialysis recovery ratio in vitro was 0.73 for all concentrations investigated. The differences between retrodialysis and dialysis recoveries were attributed to the drug's binding to the plastic tubing before and after the dialysis membrane which was experimentally evaluated and mathematically modeled. The in vivo apparent recovery determined by retrodialysis in healthy Wistar rats' kidney was 38.5+/-3.5%, similar to that observed in vitro using the same method (28.7+/-2.0%). The in vivo apparent recovery after correcting for plastic tubing binding (25.1+/-2.8%) was successfully used for determining free kidney levels of voriconazole in rats following 40 and 60mg/kg oral dosing. The results confirmed that microdialysis can be used as sampling technique to determine free tissue levels of moderately lipophilic drugs once the contribution of tubing binding and membrane diffusion on the apparent recovery are disentangled. PMID- 18395392 TI - Presence, communication and treatment of fatigue and pain complaints in incurable cancer patients. AB - OBJECTIVE: This study describes the experiences of fatigue and pain in incurable cancer patients and the treatment they receive. METHODS: Patients were recruited via medical specialists from hospitals in the South and Southwest of the Netherlands. Hundred and twenty-five incurable cancer patients filled out a written questionnaire and were also interviewed at home. RESULTS: Ninety percent reported to suffer from fatigue and 48% had pain. Forty-five percent had discussed fatigue with a healthcare professional and 55% had discussed pain. Fifteen percent reported to receive medical treatment for their fatigue and 29% received pain treatment. Treatment for fatigue and pain treatment had been recently adjusted in 4% of the patients with fatigue complaints and 21% of the patients with pain complaints. CONCLUSION: Although fatigue is a more common problem than pain in patients with incurable cancer, less attention in the care is paid to fatigue and its treatment than to pain. PRACTICE IMPLICATIONS: Fatigue deserves more attention in the care policy for incurable cancer patients and more research should be focused on interventions to address fatigue in this group of cancer patients. PMID- 18395393 TI - Patients' responsiveness to a decision support tool for primary prevention of cardiovascular diseases in primary care. AB - OBJECTIVE: Assessment of patients' responsiveness to a decision support tool for primary prevention of cardiovascular diseases (CVDs). The booklet focuses on barriers at patient level. METHODS: Process evaluation of an intervention in primary care. Patients at high or potentially high-cardiovascular risk were asked by their GP to prepare themselves for a second consultation in order to participate in decisions on risk management. OUTCOMES: Patients' actually having read the booklet and returning for the second consultation; comprehension and perceived relevance of the information; perceived reassurance. RESULTS: 17 GPs, in the intervention arm of a cluster RCT, issued 276 decision support tools during the first consultation and were instructed to ask them to return for a second consultation to discuss their CVD risk. Patients had a mean age of 54 years, 47% were male and 19% actually had a high cardiovascular risk. Data on 239 patients, a mixture of returnees and non-returnees, showed that they all read the booklet; comprehension was fair to good; 85% perceived the information as relevant; 68% of the patients felt reassured by the information. Satisfaction with the first consultation was higher in the non-returnees. CONCLUSIONS: Cardiovascular prevention spread over two consultations with use of a decision support tool for patients is not easily applicable for GPs. However, based on the findings of good patients' responsiveness, we recommend further development and implementation of decision support tools in primary care. PRACTICE IMPLICATIONS: Decision support for primary CV-prevention is welcomed by patients but needs further adjustment of both the GP and the organization of CV-prevention in primary care. Sharing information between professional and patient on a personal CV-risk management plan is difficult, more training is needed. PMID- 18395394 TI - Self-investigation to explore the impact of juvenile arthritis on adolescent life: a case-study. AB - OBJECTIVE: To gain insight into the personal experience and feelings of an adolescent with a chronic disease. METHODS: We report on the application of the self-confrontation method (SCM), illustrated by a case-example of an adolescent with juvenile idiopathic arthritis. RESULTS: Although taken at face value she was not impeded by the arthritis, through self-assessment with the SCM this adolescent acknowledged and addressed the emotional struggle to keep the arthritis secret and to constantly test the physical limits of her body. After the process of self-reflection, the adolescent showed a better integration of her arthritis experiences into her life story. CONCLUSION: With the SCM the adolescent could explore her own functioning and well-being on a manifest, as well as on an emotional and motivational level. PRACTICE IMPLICATIONS: In future research, by studying the self-investigations of a group of adolescents with chronic diseases, common risk factors for the development of a stable identity during adolescence might be identified. In clinical care, the SCM promotes self knowledge, allowing for an intrinsic motivation to deal with the emotional impact of the disease. PMID- 18395395 TI - Relationship of health behavior theories with self-efficacy among insufficiently active hypertensive African-American women. AB - OBJECTIVE: While self-efficacy plays an important role in physical activity, relatively little research has examined this construct in minorities. This study identified theoretical correlates associated with self-efficacy among insufficiently active, hypertensive Black women. METHODS: Correlates of self efficacy to: (1) overcoming barriers to physical activity; (2) making time for activity; and (3) "sticking with" physical activity were studied. RESULTS: Sixty one women (M=50.48+/-4.2 years) participated. We accounted for 32% of the variance in confidence in overcoming barriers. Women confident in overcoming barriers reported less worry about physical activity. The TTM processes of change were also in the model: consciousness raising, environmental reevaluation, counter conditioning, and self-liberation. We accounted for 16% of the variance in "making time" self-efficacy. An aversiveness barrier (e.g., physical activity is boring, physical activity is hard work) was the dominant variable in the model. Confidence to 'stick with' physical activity was associated with self reevaluation (i.e., reflection on how personal values correspond to behavior). Social support and competing demands were not associated with self-efficacy. CONCLUSIONS: Consistent with Social Cognitive Theory, results suggest that self efficacy is behavior specific and each measure likely provides unique information. PRACTICE IMPLICATIONS: Interventions should be tailored to address specific self-efficacy types. PMID- 18395396 TI - Evidence-based training of frontline health workers for door-to-door health promotion: a pilot randomized controlled cluster trial with Lady Health Workers in Sindh Province, Pakistan. AB - OBJECTIVE: Demonstrate the effective use of community-based evidence for health promotion by Lady Health Workers (LHWs) in Sindh, Pakistan. METHODS: A baseline study on mothers and children provided local evidence for risk communication tools designed and tested by LHWs. The communities were randomized to intervention and control. LHWs visited women before and after childbirth to discuss safe practices in pregnancy, in the intervention group LHW using the new tools and in the control group using their standard procedures. A household survey and focus groups permitted assessment of the impact of the intervention. RESULTS: Women in the intervention communities were more likely to attend prenatal checkups, to stop routine heavy work during pregnancy, to give colostrum to newborn babies, and to maintain exclusive breastfeeding for four months. Community focus groups confirmed a positive reaction to the tools. CONCLUSION: Discussion by lay health workers of local evidence underlying safe motherhood messages improved uptake of protective health practices. PRACTICE IMPLICATIONS: Door-to-door health promotion based on culturally appropriate interaction around relevant evidence can have a positive impact on health practices. Engaging health workers from the onset builds capacities, improves dialogue within the health system and performance of frontline health workers. PMID- 18395397 TI - Randomized controlled trial of a psychoeducation program for the self-management of chronic cardiac pain. AB - Cardiac pain arising from chronic stable angina (CSA) is a cardinal symptom of coronary artery disease and has a major negative impact on health-related quality of life (HRQL), including pain, poor general health status, and inability to self manage. Current secondary prevention approaches lack adequate scope to address CSA as a multidimensional ischemic and persistent pain problem. This trial evaluated the impact of a low-cost six-week angina psychoeducation program, entitled The Chronic Angina Self-Management Program (CASMP), on HRQL, self efficacy, and resourcefulness to self-manage anginal pain. One hundred thirty participants were randomized to the CASMP or three-month wait-list usual care; 117 completed the study. Measures were taken at baseline and three months. General HRQL was measured using the Medical Outcomes Study 36-Item Short Form and the disease-specific Seattle Angina Questionnaire (SAQ). Self-efficacy and resourcefulness were measured using the Self-Efficacy Scale and the Self-Control Schedule, respectively. The mean age of participants was 68 years, 80% were male. Analysis of variance of change scores yielded significant improvements in treatment group physical functioning [F=11.75(1,114), P<0.001] and general health [F=10.94(1,114), P=0.001] aspects of generic HRQL. Angina frequency [F=5.57(1,115), P=0.02], angina stability [F=7.37(1,115), P=0.001], and self efficacy to manage disease [F=8.45(1,115), P=0.004] were also significantly improved at three months. The CASMP did not impact resourcefulness. These data indicate that the CASMP was effective for improving physical functioning, general health, anginal pain symptoms, and self-efficacy to manage pain at three months and provide a basis for long-term evaluation of the program. PMID- 18395398 TI - Sex differences in the adequacy of pain management among patients referred to a multidisciplinary cancer pain clinic. AB - Few studies have evaluated sex differences in the adequacy of pain management in cancer. Existing studies have been marked by methodological limitations and results have been mixed. The present study sought to determine whether sex was associated with pain severity and pain management in cancer patients newly referred by their primary oncology team to a multidisciplinary cancer pain clinic. One hundred thirty-one cancer patients completed the Brief Pain Inventory Short Form and medical chart review was conducted to obtain patients' clinical characteristics and pain treatment data. There were no differences between males and females in ratings of worst pain in the last week. Females were significantly less likely to have been prescribed high potency opioids by their primary oncology team and significantly more likely to report inadequate pain management as measured by Pain Management Index scores. These results suggest a sex bias in the treatment of cancer pain and support the routine examination of the effect of sex in cancer pain research. PMID- 18395399 TI - Nonconvulsive status epilepticus in terminally ill patients-a diagnostic and therapeutic challenge. AB - Nonconvulsive status epilepticus (NCSE) is rarely considered as a cause of confusion in terminally ill patients. The clinical presentation varies from altered mental status to coma, without visible convulsions. Electroencephalogram is the most important diagnostic tool to identify epileptiform activity. Treatment should be initiated following a stepwise model, avoiding intubation and transfer to the intensive care unit. Although mortality rates are high, in some patients NCSE can be reversed by treatment. NCSE should be considered in a differential diagnosis of all terminally ill patients with sudden mental status changes. We present two cases and discuss treatment options and the borders of therapy. PMID- 18395400 TI - Regulating palliative care: the case of hospice. AB - Palliative care services provided to patients and families vary substantially across hospices. Literature suggests regulation can act as a standardizing force in health care delivery. However, little is known about the effect of regulation on the delivery of palliative care in hospice and whether its effect differs for different types of hospice providers. We estimated the association between regulation, defined as Medicare hospice certification, and the delivery of palliative care in hospice using a nationally representative data set of 9,409 patients from 2,066 hospices surveyed in the National Home and Hospice Care Survey, 1992-2000. Using multivariable analysis, we found Medicare hospice certification was associated with a significantly broader range of services provided to patients (odds ratio [OR]=2.45; 95% confidence interval [CI]: 1.16, 5.17). This effect was significantly more pronounced (P-value for interaction=0.001) among for-profit hospices (OR=15.24; 95% CI: 4.06, 57.17) than among nonprofit hospices (OR=1.53; 95% CI: 0.75, 3.14). The effect of ownership on certification differences was most apparent for the provision of skilled nursing (prevalence difference in difference=52.4%), spiritual care (prevalence difference in difference=49.6%), and social services (prevalence difference in difference=48.1%). This study is the first to demonstrate the substantial association between the regulation of hospices and the provision of a multidisciplinary range of services to patients and families. It provides valuable insights regarding the potential role of regulation in standardizing the quality of palliative care across the increasingly diverse palliative care programs developing outside of hospice. PMID- 18395401 TI - Hospice and palliative care development in India: a multimethod review of services and experiences. AB - Palliative care has been developing in India since the mid-1980s, but there is a dearth of evidence about service provision on which to base national policy and practice. The aim of this study was to assess the current state of palliative care in India, mapping the existence of services state by state, and documenting the perspectives and experiences of those involved. A multimethod review was used, which included synthesis of evidence from published and grey literature, ethnographic field visits, qualitative interviews with 87 individuals from 12 states, and collation of existing public health data. The review identified 138 hospice and palliative care services in 16 states and union territories. These are mostly concentrated in large cities, with the exception of Kerala, where they are much more widespread. Nongovernmental organizations, public and private hospitals, and hospices are the predominant sources of provision. We were unable to identify palliative care services in 19 states/union territories. Development of services is uneven, with greater provision evident in the south than the north, but for the majority of states, coverage is poor. Barriers to the development of palliative care include: poverty, population density, geography, opioid availability, workforce development, and limited national palliative care policy. Successful models exist for the development of affordable, sustainable community-based palliative care services. These have arisen from adapting Western models of hospice and palliative care for implementation in the Indian cultural context. Further work is required to ensure that the growing interest in hospice and palliative care in India is used to increase the momentum of progress. PMID- 18395402 TI - Assessment of xerostomia and its impact on quality of life in head and neck cancer patients undergoing radiotherapy, and validation of the Taiwanese version of the xerostomia questionnaire. AB - The purposes of this study were to (a) explore the impact of xerostomia and saliva flow on quality of life and (b) validate the Taiwanese version of the Xerostomia Questionnaire (XQ) for patients undergoing radiotherapy (RT) for head and neck cancer in Taiwan. This was a prospective longitudinal study. Instruments consisted of the Xerostomia Questionnaire-Taiwan version (XQ-T) and the Medical Outcomes Study Short Form-36 Taiwan Version. Salivary output was measured by collecting unstimulated whole saliva. The questionnaires and measurements of salivary output were completed before RT was initiated and at two, four, six, and eight weeks after RT had started. Changes in xerostomia scores, quality of life, saliva flow, and predictors of quality of life over time were examined by using general estimating equations. The XQ-T is the first xerostomia measurement instrument developed for use with Taiwanese cancer patients and demonstrated excellent reliability and validity. Saliva flow was significantly correlated with XQ-T scores at two, four, six, and eight weeks after RT had started, but not before RT had begun. Saliva flow and quality-of-life scores significantly diminished and xerostomia scores significantly increased over the eight-week period. Saliva flow and XQ-T scores significantly predicted quality of life, after adjusting for the maturation effect. The results of this study show that the XQ-T is the first xerostomia measurement instrument to be developed for Taiwanese cancer patients and demonstrates excellent reliability and validity. PMID- 18395403 TI - Magnesium sulfate via a mucous fistula in the management of short bowel syndrome. PMID- 18395404 TI - Palliative percutaneous nephrostomy in recurrent cervical cancer: a retrospective analysis of 50 consecutive cases. AB - Cervical cancer is a public health problem in Brazil, with annual incidence rates of 20-40 cases/100,000 women. Most patients with recurrent disease have symptoms from locoregional disease and may develop renal failure. This study aims to evaluate the outcome of patients with recurrent cervical cancer who underwent percutaneous nephrostomy (PN). We reviewed the medical records of 50 such patients who were referred to the Palliative Care Unit of the Brazilian National Cancer Institute from January 2002 to October 2006. Median age was 44 years (range, 26-67 years). Half the patients had improvement in pain or uremic symptoms, and seven (14%) had improved performance status (PS) after the procedure. Thirty patients (60%) had improvement of renal function; median creatinine levels before and after PN were 6.4 and 3.7mg/dL, respectively (P<0.05). Median overall survival after PN was 8.9 weeks (95% confidence interval [CI]: 7.4-10.3). Median survival was 9.9 weeks (95% CI: 8.7-11.0) in 40 patients with baseline PS 1-3 and one week (95% CI: 0.1-1.9) in 10 patients with PS 4 (log rank, P<0.0001). Median survival in patients with and without improvement of renal function after PN was 10.0 weeks (95% CI: 8.6-11.3) and 2.6 weeks (95% CI: 0-11.3), respectively (log rank, P=0.01). Twenty-nine patients (58%) died from renal failure. Complications were mainly urinary tract infection (n=10), catheter loss (n=9), and bleeding (n=1). These data suggest that PN can be of clinical benefit for carefully selected patients with recurrent cervical cancer. PMID- 18395405 TI - Persistent activation of Akt or ERK prevents the toxicity induced by saturated and polyunsaturated fatty acids in RINm5F beta-cells. AB - The aim of this study was to investigate whether the toxicity of saturated and polyunsaturated fatty acids (PUFA) on RINm5F cells is related to the phosphorylation state of Akt, ERK and PKC delta. The regulation of these kinases was compared in three experimental designs: (a) 4h-exposure, (b) 4h-exposure and a subsequent withdrawn of the FA for a 20 h period and (c) 24h-exposure. Saturated and PUFA were toxic to RINm5F cells even at low concentrations. Also, evidence is provided for a late (i.e. the effect only appeared hours after the treatment) and a persistent regulation (i.e. maintenance of the effect for several hours) of Akt, ERK and PKC delta phosphorylation by the FA. Late activation of PKC delta seems important for palmitate cytotoxicity. Persistent activation of the survival proteins Akt and ERK by stearate, oleate and arachidonate might play an important role to prevent the toxic effect of posterior PKC delta activation. The results shown may explain why a short-period exposure to FA is not enough to induce cytotoxicity in pancreatic beta-cells, since survival pathways are activated. Besides, when this activation is persistent, it may overcome a posterior induction of death pathways. PMID- 18395406 TI - Gene expression profiling of in vitro cultured macrophages after exposure to the respiratory sensitizer hexamethylene diisocyanate. AB - Occupational exposure to chemicals is one of the main causes of respiratory allergy and asthma. Identification of chemicals that trigger allergic asthma is difficult as underlying processes and specific markers have not yet been clearly defined. Moreover, adequate classification of the respiratory toxicity of chemicals is hampered due to the lack of validated in vivo and in vitro test methods. The study of differential gene expression profiles in appropriate human in vitro cell systems is a promising approach to identify selective markers for respiratory allergy. As alveolar macrophages display important immunological and inflammatory properties in response to foreign substances in the lung, we aimed at gaining more insight in changes of human macrophages transcriptome and to identify selective genetic markers for respiratory sensitization in response to hexamethylene diisocyanate (HDI). In vitro cultures of human THP-1 cells were differentiated into macrophages and exposed to 55 microg/ml HDI for 6 and 10h. Using human oligonucleotide microarrays, changes were observed in the expression of genes that are involved in diverse biological and molecular processes, including detoxification, oxidative stress, cytokine signaling, and apoptosis, which can lead to the development of asthma. These genes are possible markers for respiratory sensitization caused by isocyanates. PMID- 18395407 TI - Screening for neuropsychological impairment using Reitan and Wolfson's preliminary neuropsychological test battery. AB - This study examined the utility of a screening battery developed by Reitan & Wolfson, 2006 for predicting neuropsychological impairment on the Halstead-Reitan neuropsychological test battery for adults. Using archival neuropsychological data from 69 litigants seen in a private practice setting, the Pearson correlation between the General Neuropsychological Deficit Scale (GNDS) score and the total Neuropsychological Deficit Scale (NDS) score from the screening battery (SBNDS) was .82. ROC curve analysis determined that the AUC was .88. Using a cutoff score of 9, as recommended by Reitan and Wolfson, the screening battery had excellent specificity but only fair sensitivity for identifying individuals with neuropsychological impairment on the Halstead-Reitan battery. Using a cutoff score of 8, the sensitivity and specificity of the screening battery was comparable to the findings of Reitan and Wolfson. The findings from this study indicate the optimal cutoff score for the screening battery may vary with different populations. The positive predictive power (PPP) and negative predictive power (NPP) were calculated for various base rates for cut scores with both sensitivity and specificity of greater than .600, and this information is provided. PMID- 18395408 TI - Exploring the utility of the Spence Children's Anxiety Scales parent- and child report forms in a North American sample. AB - This study examined the psychometric properties of the Spence Children's Anxiety Scale child- and parent-report forms in a sample of 85 children from the community and 85 children with anxiety disorders matched on sex and age. Results indicated that both versions of the scale had good internal reliability, with the exception of the physical injury fears subscale. All subscales and the total scores of both versions discriminated children in the anxious group from community controls. Strong correlations between child and parent reports supported the concurrent validity of the measure. Furthermore, convergent and divergent validity of the scales were supported by significant correlations with negative affect and physiological hyperarousal to a greater extent than with positive affect indicating that SCAS and SCAS-P measured anxiety rather than depressive symptoms. Mean differences with research from other countries were apparent for selected subscales and supported the need for culture-specific norms. PMID- 18395409 TI - Evaluation of intolerance of uncertainty: development and validation of a new self-report measure. AB - Intolerance of uncertainty represents an important risk factor for development of anxiety disorders. However, few measures have been developed in order to evaluate this construct. Four studies were conducted in order to validate a new instrument evaluating intolerance of uncertainty: the Intolerance of Uncertainty Inventory (IUI). The first study described the questionnaire's development and evaluated the psychometric properties of its preliminary version. Study 2 examined the reliability and the factorial validity of the final version of the questionnaire, while Study 3 mainly addressed its convergent validity. Finally, Study 4 examined the questionnaire's temporal stability. Factorial analyses confirmed the IUI's validity. Results also supported the IUI's reliability, convergent validity, and temporal stability. The IUI is the first instrument that offers the possibility of measuring intolerance of uncertainty as a tendency to consider uncertainties to be unacceptable, as well as in terms of cognitive and behavioral manifestations. PMID- 18395410 TI - Coping style and immunity in animals: making sense of individual variation. AB - Predicting the individual vulnerability to immune mediated disease is one of the main challenges of modern biomedical research. However, the question of individual behavioral and physiological characteristics that might predict this vulnerability has been subject of research and debate for a long time. This paper will argue that animal models aimed at individual vulnerability should consider the biological function of variation in nature. An increasing number of studies show the ecological significance of variation within a species. Based on behavioral studies in several vertebrate species two coping style can be distinguished. Variation in coping style appears to play a role in the population dynamics and the evolutionary fitness of the species. Coping styles are reflected in a stable differentiation in the behavioral and physiological stress responsiveness over time and across situations. Based on the observations that the individual level of offensive aggressive behavior (i.e., the tendency to defend the home territory) is strongly related to the way animals react to various other environmental challenges, it is argued that the individual's level of offensiveness is an important indicator and component of a more trait-like behavioral and physiological response pattern (coping style) to environmental demands. The coping style of aggressive animals is principally aimed at a (pro)active prevention or manipulation of a stressor whereas the non-aggressive individuals tend to passively accept or react to it. Proactive coping is associated with high sympathetic reactivity to stressors whereas the more passive or reactive coping style generally has a higher HPA axis reactivity. In view of the immune modulating nature of these major neuroendocrine stress systems, one might expect that coping styles will be reflected in a differential vulnerability to immune mediated disease as well. Indeed, several studies have demonstrated such a relationship, indicating that the functional variation in coping style and related neuroendocrine stress reactivity, as it occurs in nature, might be a good standard for studies aimed at understanding individual vulnerability. This is in agreement with more recent views that also in humans stress reactivity may be the best predictor for the individual vulnerability to immune mediated diseases. This asks for a more fundamental and translational approach of individual disease vulnerability based on a common biological basis of individual differentiation in behavior and physiology in humans and animals. PMID- 18395411 TI - Myeloproliferative disorder as the cause of peripheral ischemia in a young patient. AB - A 35-year-old patient with rest pain and peripheral gangrene was referred with a diagnosis of Takayasu's disease but had myeloproliferative disorder (primary thrombocythemia). The angiogram revealed surgically noncorrectable small vessel occlusive disease with no evidence of atheroma. Medical therapy with cytotoxic and antiplatelet agents achieved satisfactory results with excellent wound healing after surgical amputation of necrotic tissue. The need for early diagnosis of this condition is emphasized. PMID- 18395412 TI - A novel molecular mechanism to account for the action of resveratrol against reperfusion injury. PMID- 18395413 TI - Spontaneous early recanalization of an acute symptomatic critical stenosis of the extracranial internal carotid artery: a case report. AB - We present a 63-year-old female with critical stenosis of the extracranial portion of the internal carotid artery (ICA), diagnosed using color-coded Duplex ultrasonography (CCDU) and magnetic resonance imaging. Nine days later, the patient showed profound clinical improvement, at which time spontaneous recanalization of the previously highly stenosed ICA was seen in follow-up CCDU and magnetic resonance angiography (MRA) and confirmed by three-dimensional computed tomography angiography (CTA). The detection of recanalization is important in predicting the patient's prognosis and deciding a suboptimal secondary prophylaxis strategy of medical or surgical treatment. Therefore, we suggest performing imaging studies immediately in patients, especially potential surgical or stenting candidates showing great changes in neurological function, through examination with CCDU and confirmation with MRA or CTA, all safe and minimally invasive methods, to see whether recanalization has occurred. PMID- 18395415 TI - Simple procedures to obtain exogenous internal controls for use in RT-PCR detection of bovine pestiviruses. AB - Pestiviruses are ubiquitous pathogens of cattle and frequent adventitious viruses in biologicals. Furthermore, it has been suggested that these agents might be related to infantile gastroenteritis and microencephaly. Since the virus is highly prevalent in fetal bovine serum, the risk of contamination is high in most laboratories. Thus, the implementation of detection methods in all laboratories is of worth. Despite continuous surveillance, these agents have been detected in cell lines, fetal bovine serum, live and inactivated animal and human vaccines and interferon for human use. In this report, DNA and RNA internal controls (ICs) which can be implemented in laboratories with minimal equipment are described. The developed standards can be added before RNA purification, allowing to monitor all steps of the protocol (viral RNA extraction, reverse transcription and cDNA amplification). It is shown that inhibitory effects that could lead to decreased sensitivity can be minimized by controlling the amount of mimic molecules added to the samples. A method to avoid the problem of DNA traces present in in vitro transcribed RNA preparations is provided. PMID- 18395414 TI - Value of cutting balloon angioplasty for limb salvage in patients with obstruction of popliteal and distal arteries. AB - Popliteal-to-distal bypass is still the gold standard for limb salvage. However, some patients, especially elderly or diabetic patients, are not eligible for such treatment; and problems may arise, including poor healing of distal surgical wounds, delayed resumption of ambulation, and prolonged hospitalization. This prospective multicenter study carried out on an intent-to-treat basis includes 53 extremities in 48 patients presenting critical ischemia due to infrageniculate arterial lesions with no proximal lesions. Two populations were isolated: diabetic patients (56.6%) and elderly patients over 80 years (45%). In 82% of cases the arterial lesions were long, i.e., more than 1 cm. The limb salvage rate at 1 year was 81%. Postoperative mortality was 9%, and mortality at 1 year was 22.6%. These results show that cutting balloon angioplasty can be proposed as primary treatment in patients with critical ischemia due to popliteal and distal artery lesions. PMID- 18395416 TI - Can social stories enhance the interpersonal conflict resolution skills of children with LD? AB - Since many children with learning disabilities (LD) face interpersonal conflict resolution problems, this study examines the efficacy of social stories in helping them choose more appropriate interpersonal conflict resolution strategies. A social story was recorded and played to the 31 children with LD in the experimental group twice a week for a period of 1 month, while the 32 children with LD in the control group did not receive any intervention. The effects of the intervention were systematically examined by means of an interview with the participants, while teachers completed the T-MESSY (Matson, J. L. (1990). Matson Evaluation of Social Skills With Youngsters: Manual. Worthington, OR: International Diagnostic Systems). All children chose mainly avoidance and hostile strategies before the intervention, but children in the experimental group chose predominantly positive strategies both after the intervention and at follow-up in comparison to control children. Furthermore, children with LD who received the intervention were rated by their teachers as engaging in significantly less inappropriate social behaviors after the intervention and at follow-up in comparison to control children. The recorded changes in the choice of interpersonal conflict resolution strategies and the more positive teacher ratings for the experimental group indicate that social stories constitute a powerful intervention for the enhancement of the social competence of children with LD. PMID- 18395417 TI - Laryngoaltimeter: a new ambulatory device for laryngeal height control, preliminary results. AB - The vertical larynx position has significant effects on vocal tract resonances and on the biomechanical properties of the vocal folds. It is generally agreed that the larynx should be kept in a comfortably low position during singing, and voice problems are often associated with a habitually raised larynx. A new method, and a battery operated portable device called Laryngoaltimeter, was developed to control laryngeal height continuously for therapeutic or educational purposes. Two similar condenser microphones attached on the suprasternal notch and supraglottic region were used to capture corresponding vibrations during phonation. The microphone signals were then filtered through a band pass filter (90-240 Hz), digitized, and compared to each other by detection algorithms of the Laryngoaltimeter. When the supraglottic microphone received subglottic resonance related vibrations due to laryngeal elevation, auditory and visual signals were produced as a biofeedback by the device. Waterfall amplitude spectrograms of the microphone signals verified that the frequencies captured by each microphone were dissimilar before and similar after laryngeal elevation. The accuracy of the device was found to be 87% on 13 subjects having different voice classifications as a demonstration of its use. Laryngoaltimeter is a prototype device and needs to be developed. Further research may also be established to investigate the habitual effects of keeping vertical position low by using Laryngoaltimeter as a biofeedback device during therapy exercises or voice lessons. PMID- 18395418 TI - Predicted singers' vocal fold lengths and voice classification-a study of x-ray morphological measures. AB - Students admitted to the solo singing education at the University of Music Dresden, Germany have been submitted to a detailed physical examination of a variety of factors with relevance to voice function since 1959. In the years 1959 1991, this scheme of examinations included X-ray profiles of the singers' vocal tracts. This material of 132 X-rays of voice professionals was used to investigate different laryngeal morphological measures and their relation to vocal fold length. Further, the study aimed to investigate if there are consistent anatomical differences between singers of different voice classifications. The study design used was a retrospective analysis. Vocal fold length could be measured in 29 of these singer subjects directly. These data showed a strong correlation with the anterior-posterior diameter of the subglottis and the trachea as well as with the distance from the anterior contour of the thyroid cartilage to the anterior contour of the spine. These relations were used in an attempt to predict the 132 singers' vocal fold lengths. The results revealed a clear covariation between predicted vocal fold length and voice classification. Anterior-posterior subglottic-tracheal diameter yielded mean vocal fold lengths of 14.9, 16.0, 16.6, 18.4, 19.5, and 20.9mm for sopranos, mezzo-sopranos, altos, tenors, baritones, and basses, respectively. The data support the assumption that there are consistent anatomical laryngeal differences between singers of different voice classifications, which are of relevance to pitch range and timbre of the voice. PMID- 18395419 TI - The frequency of perceived stress, anxiety, and depression in patients with common pathologies affecting voice. AB - The study's objectives were to investigate (1) the frequency of perceived stress, anxiety, and depression for patients with common voice disorders, (2) the distribution of these variables by diagnosis, and (3) the distribution of the variables by gender. Retrospective data were derived from self-report questionnaires assessing recent stress (Perceived Stress Scale-10), anxiety, and depression (Hospital Anxiety and Depression Scale) in a cohort of new patients presenting to a voice clinic. Data are presented on 160 patients with muscle tension dysphonia (MTD), benign vocal fold lesions, paradoxical vocal fold movement disorder (PVFMD), or glottal insufficiency. Pooled data indicated that average stress, anxiety, and depression scores were similar to those found for the healthy population. However, 25.0%, 36.9%, and 31.2% of patients showed elevated stress, anxiety, and depression scores, respectively, compared to norms. Patients with PVFMD had the most frequent occurrence-and patients with glottal insufficiency had the least frequent occurrence of elevated stress, anxiety, and depression. Stress and depression were more common with MTD than with lesions, whereas reverse results were obtained for anxiety. More females than males had elevated stress, anxiety, and depression scores. The data are consistent with suggestions that stress, anxiety, and depression may be common among some patients with PVFMD, MTD, and vocal fold lesions and more common for women than men. However, individual variability in the data set was large. Further studies should evaluate the specific role of these conditions for selected categories of voice disorders in susceptible individuals. PMID- 18395420 TI - Predictors of phoneme and stress perception in undergraduate students of singing. AB - The purpose of this exploratory study was to examine the relationship between undergraduate vocal music majors' diction acquisition abilities for singing in a nonnative language (as rated both by themselves and by their studio voice teachers) and their scores on an objective test of phonemic and stress perception. Ten students with varying levels of university voice training served as participants. The results showed significant negative correlations between each of the teachers' four ratings and the students' scores on the phonemic awareness subtest. In addition, 20% of the students demonstrated evidence of underdeveloped phonemic awareness skills, as indicated by their below average test performance. Considerable individual differences were also observed in the students' abilities to track phonemes within a sequence of phonemes, count and track syllables within a sequence of syllables, and track combinations of phoneme and syllable changes in sequence, as evidenced by subtest performance scores. These findings corroborate existing reports which indicate that approximately 30% of the population does not fully develop phonemic awareness skills in the absence of special training. The findings support the utility of this objective test of phonemic and stress perception as a means of identifying students who will have difficulty with diction acquisition, and point to possibilities for pretraining to improve their response to diction instruction. PMID- 18395421 TI - Histologic characterization of human scarred vocal folds. AB - Vocal fold scarring remains a significant problem. Although several animal models have been developed to improve our understanding of the histopathology, the histologic features of scarred human vocal folds have rarely been reported. The present case studies aimed to define the histologic changes of scarred human vocal folds caused by cordectomy or cordotomy. Ten patients with the scarred vocal folds were involved in this study. Nine patients with early glottic cancer underwent endoscopic cordectomy, and one patient underwent superficial cordotomy for idiopathic scar. The postcordectomy or cordotomy scar was biopsied or resected 3-13 months after the original procedure. After confirming absence of any tumor in cancer patients, the remaining specimens were used in the present study. Histologic examination investigated deposition of extracellular matrix (ECM) including collagen, elastin, hyaluronic acid (HA), fibronectin, and decorin in the lamina propria of the scarred vocal folds. There was a wide range of variation in the deposition of ECM in scarred vocal folds. Excessive and disorganized collagen deposition was observed in most cases that had undergone deep resection of the lamina propria, whereas deposition of collagen was mild and well organized after superficial resection. Decorin was retained in all cases after superficial cordectomy or cordotomy, but varied after deep resection. Deposition of elastin, HA, and fibronectin varied regardless of depth of injury. Histology of scarred vocal folds may vary with degree of injury and individual healing mechanism. PMID- 18395422 TI - Endogenous Gas6 and Ca2+ -channel activation modulate phagocytosis by retinal pigment epithelium. AB - Mutation or loss of MerTK as well as deficiency of alphavbeta5-integrins, gives rise to retinal-degeneration due to inefficient phagocytosis of photoreceptor outer-segment fragments by the retinal pigment epithelium (RPE). This study shows that Gas6 expressed endogenously by human RPE promotes phagocytosis. The RPE expresses Gas6 more highly in vivo and in serum-reduced conditions in vitro than in high-serum conditions, suggesting a negative-feedback control. An antibody blockage approach revealed that Gas6-expressing RPE phagocytizes photoreceptor outer-segment fragments due to stimulation of MerTK by endogenous Gas6 in vitro. MerTK- and Gas6-antibodies reduced phagocytosis. Blocking L-type Ca(2+)-channels with nifedipine inhibited MerTK dependent phagocytosis in vitro. Application of integrin inhibitory, soluble, RGD-containing peptides or soluble vitronectin reduced L-type Ca(2+)-channel currents in RPE. Herbimycin A, which reduces phosphorylation of integrin receptor-associated proteins and decreases L-type Ca(2+)-channel currents in RPE, eliminates the inhibiting vitronectin effect and abolishes phagocytosis. Thus, Gas6-promoted phagocytosis was inhibited by L-type Ca(2+)-channel blockage, which in turn may be activated by integrin receptor stimulation. These results suggest that L-type Ca(2+)-channels could be regulated downstream of both MerTK and alphavbeta5-integrin, indicating that the binding and uptake mechanisms of phagocytosis are part of a converging pathway. PMID- 18395423 TI - Different kinases desensitize the human delta-opioid receptor (hDOP-R) in the neuroblastoma cell line SK-N-BE upon peptidic and alkaloid agonists. AB - In a previous work, we described a differential desensitization of the human delta-opioid receptor (hDOP-R) by etorphine (a non-selective and alkaloid agonist) and delta-selective and peptidic agonists (DPDPE ([D Pen(2,5)]enkephalin) and deltorphin I (Tyr-D-Ala-Phe-Asp-Val-Val-Gly-NH(2))) in the neuroblastoma cell line SK-N-BE (Allouche et al., Eur. J. Pharmacol., 371, 235, 1999). In the present study, we explored the putative role of different kinases in this differential regulation. First, selective chemical inhibitors of PKA, PKC and tyrosine kinases were used and we showed a significant reduction of etorphine-induced opioid receptor desensitization by the bisindolylmaleimide I (PKC inhibitor) while genistein (tyrosine kinase inhibitor) was potent to impair desensitization induced by the different agonists. When the PKA was inhibited by H89 pretreatment, no modification of opioid receptor desensitization was observed whatever the agonist used. Second, we further studied the role of G protein coupled receptor kinases (GRKs) and by using western-blot experiments we observed that only the GRK2 isoform was expressed in the SK-N-BE cells. Next, the neuroblastoma cells were transfected with the wild type GRK2 or its dominant negative mutant GRK2-K220R and the inhibition on cAMP level was determined in naive and agonist-pretreated cells. We showed that over-expression of GRK2-K220R totally abolished etorphine-induced receptor desensitization while no effect was observed with peptidic agonists and over-expression of GRK2 selectively impaired cAMP inhibition promoted by etorphine suggesting that this kinase was involved in the regulation of hDOP-R activated only by etorphine. Third, correlation between functional experiments and phosphorylation of the hDOP-R after agonist activation was assessed by western-blot using the specific anti-phospho-DOP-R Ser(363) antibody. While all agonists were potent to increase phosphorylation of opioid receptor, we showed no impairment of receptor phosphorylation level after PKC inhibitor pretreatment. Upon agonist activation, no enhancement of receptor phosphorylation was observed when the GRK2 was over-expressed while the GRK2 K220R partially reduced the hDOP-R Ser(363) phosphorylation only after peptidic agonists pretreatment. In conclusion, hDOP-R desensitization upon etorphine exposure relies on the GRK2, PKC and tyrosine kinases while DPDPE and deltorphin I mediate desensitization at least via tyrosine kinases. Although the Ser(363) was described as the primary phosphorylation site of the mouse DOP-R, we observed no correlation between desensitization and phosphorylation of this amino acid. PMID- 18395424 TI - Orbitofrontal, amygdala and hippocampal volumes in teenagers with first presentation borderline personality disorder. AB - It is not known whether the fronto-limbic volume reductions found in adults with established borderline personality disorder (BPD) are present early in the disorder. The aim of the study was to investigate orbitofrontal cortex (OFC), hippocampal and amygdala volumes in a first-presentation teenage BPD sample with minimal exposure to treatment. Groups of 20 BPD patients and 20 healthy control participants underwent magnetic resonance imaging. Hippocampal, amygdala, OFC and whole brain volumes were estimated and compared between the two groups. Analysis of variance revealed reversal of the normal (right>left) asymmetry of OFC grey matter volume in the BPD group, reflecting right-sided OFC grey matter loss in the BPD group compared with control participants. No significant differences were found for amygdala or hippocampal volumes comparing BPD with control participants. We identified OFC but not hippocampal or amygdala volumetric differences early in the course of BPD. Hippocampal and amygdala volume reductions observed in adult BPD samples might develop during the course of the disorder, although longitudinal studies are needed to examine this. PMID- 18395426 TI - Abstracts of 19th Helsinki Drug Research 2008. June 9-11, 2008. Helsinki, Finland. PMID- 18395425 TI - Interaction of nicotine with magnesium aluminum silicate at different pHs: characterization of flocculate size, zeta potential and nicotine adsorption behavior. AB - The purposes of this study were to prepare dispersions in various ratios of nicotine (NCT) and magnesium aluminum silicate (MAS) at different pHs and to investigate interaction of NCT with MAS by characterizing microscopic morphology, particle size and zeta potential of MAS-NCT flocculates. Moreover, the NCT adsorption onto MAS at different pHs were also investigated. At basic medium, incorporating NCT into MAS dispersion brought about a small decrease in the zeta potential of MAS, leading to a loose flocculate formation of MAS. This is likely to be due to an adsorption of unionized form of NCT onto MAS via intermolecular hydrogen bonding. The lower zeta potential, denser matrix structure and larger size of the flocculates was found at neutral and acidic media because the protonated species of NCT could interact with the negatively charged MAS by electrostatic force. In addition the flocculates formed at pH 4 possibly possessed a higher density than those formed at pH 7, suggesting that the diprotonated species of NCT at pH 4 caused stronger interaction with MAS. The adsorption isotherms of NCT onto MAS at different pHs can be described not only using the Langmuir model, but also using the Freundlich model. The higher affinity of NCT adsorption onto MAS at neutral and acidic media was found. However, the adsorption capacity to form NCT monolayer reduced with decreasing the pH of the dispersions because of an adsorption of hydronium ions and a decrease in surface area of adsorption site by flocculation. These findings suggested that the flocculation of MAS dispersion could be induced by incorporation of NCT. The characteristics, such as particle size and zeta potential, of the NCT-MAS flocculates and the adsorption isotherms of NCT onto MAS were depended upon pH of dispersion, in which the different charged species of NCT were formed. PMID- 18395427 TI - Long-term survival and late toxicity after chemoradiotherapy for cervical cancer- the Addenbrooke's experience. AB - AIM: To evaluate the long term cause-specific survival and late toxicity of chemoradiotherapy for carcinoma of the cervix treated outside research settings. MATERIALS AND METHODS: Between May 1999 and April 2003, 74 patients with carcinoma of the cervix were treated with radical radiotherapy given concurrently with weekly cisplatin chemotherapy. Three patients died during treatment, leaving 71 patients available for analysis of long-term survival and late toxicity of treatment. In total, 56 patients (78.9%) received chemoradiotherapy as primary radical treatment. Ten patients (14.1%) received chemoradiotherapy as adjuvant treatment after radical surgery. The remaining five patients (7.0%) received chemoradiotherapy as salvage treatment for pelvic recurrences after previous surgery. Forty-seven (66.2%) patients had squamous cell carcinomas, whereas 24 (33.8%) patients had adenocarcinomas. RESULTS: The median follow-up for surviving patients was 64 months. The actuarial 5-year cause-specific survival for the 66 patients undergoing primary treatment (chemoradiotherapy+/-surgery) was 54.6%. The cause-specific survival by International Federation of Gynecology and Obstetrics (FIGO) disease stage was 58.3% for stage I disease, 69.9% for stage II disease and 20.8% for stage III disease. The actuarial 5-year pelvic control rate for the same group of patients was 73.3% overall (stage I=79.2%, stage II=89.0%, stage III=33.3%). Four of the five patients treated for recurrent disease are alive and well with a median follow-up of 70 months. Of the 66 patients undergoing primary treatment, seven (10.6%) had persistent disease after chemoradiotherapy. Of the 22 patients (33.3%) who relapsed >6 months after treatment, eight (36.4%) relapsed within the pelvis alone, 12 (54.5%) had metastatic disease alone, whereas two (9.1%) had both local and distant relapse. The overall rates of pelvic and distant relapse were 25.8 and 21.2%, respectively. Eight of 23 patients (34.8%) with adenocarcinomas developed metastatic disease compared with only six of 43 patients (14.0%) with squamous cell tumours. Thirteen patients (18.3%) had at least one complication that was classified as grade 3 or 4. Six patients (8.5%) had grade 3 or 4 urinary complications, five (7.0%) had grade 3 or 4 bowel complications and six (8.5%) had grade 3 or 4 complications affecting other organs. Five patients had grade 3 or 4 complications affecting more than one organ. The actuarial rate for grade 3 or 4 urinary complications was 14.5%, 9.4% for grade 3 or 4 bowel complications and 11.4% for grade 3 or 4 complications affecting other organs. The overall actuarial risk for grade 3 or 4 long-term morbidity in the study group was 28.2%. There were no significant correlations between the incidence of serious late toxicity and disease stage, field arrangement, treatment volumes or postoperative radiotherapy. CONCLUSIONS: Our study has shown that the addition of chemotherapy to radiotherapy for cervical cancer probably improves the survival of patients treated outside research settings, but the benefit may not be as large as that obtained in clinical trials and the risk of serious late toxicity is increased. Further developments to improve survival and local control and to minimise toxicity are therefore necessary. PMID- 18395428 TI - Socio-economic differences in the prevalence of diabetes in Italy: the population based Turin study. AB - BACKGROUND AND AIMS: A surveillance programme on diabetes based on administrative data is being implemented in the city of Turin, Italy. The aim of this study is to assess socio-economic differences in the prevalence of diabetes in this large Italian population-based cohort. METHODS AND RESULTS: People with known diabetes resident in Turin on July 31, 2003 were identified through three data sources: the regional register of persons with diabetes, hospital discharges and prescriptions for antidiabetic drugs. Data sources were linked to the Turin population register to obtain individual data on educational level and census tract median income. Missing cases were estimated by using the capture-recapture method. We identified 34,420 persons with diabetes; prevalence adjusted for undercount was 4.91% (95% Confidence Intervals: 4.69-5.22) among men and 4.68% (4.41-5.08) among women. Age adjusted prevalence ratios between low and high educational levels were 2.32 (2.23-2.41) in men, and 3.45 (3.28-3.62) in women. Social inequalities were larger in women than in men and in people aged 21-65 years than in those age >65 years. CONCLUSION: This population-based study shows that there are socio-economic inequalities in the prevalence of the disease, particularly in women, and in young people. Our findings indicate that: (1) prevention of diabetes should be mainly focused on the socially disadvantaged strata of the population; (2) a low cost surveillance programme of diabetes using routinely collected data is feasible to better assist public health policies. PMID- 18395429 TI - Sarcopenic obesity: a new category of obesity in the elderly. AB - BACKGROUND AND AIM: In elderly patients, age-related changes in body composition, as well as the increased prevalence of obesity, determine a combination of excess weight and reduced muscle mass or strength, recently defined as sarcopenic obesity (SO). This review examines the main studies regarding sarcopenic obesity in the elderly. DATA SYNTHESIS: Definition of SO necessarily combines those of sarcopenia and obesity. The prevalence of sarcopenia and SO increases with age. Muscle and fat mass are strongly interconnected from a pathogenetic point of view. A better understanding of the mechanisms which lead from loss of muscle mass to fat gain or vice versa from fat gain to muscle loss seems to be crucial. Recent data suggest that peptides produced by adipose tissue may play an important role in the pathophysiology of SO, thus more research is needed to better characterize this new area. Obesity and sarcopenia in the elderly may potentiate each other maximizing their effects on disability, morbidity and mortality. Identifying elderly subjects with SO should be mandatory; effective treatment of sarcopenia and SO may attenuate its clinical impact. CONCLUSION: The concept of SO may help to clarify the relationship between obesity, morbidity and mortality in the elderly. PMID- 18395430 TI - Specific permeability modulation of intestinal paracellular pathway by chitosan poly(isobutylcyanoacrylate) core-shell nanoparticles. AB - This work is focused on the evaluation of the in vitro permeation modulation of chitosan and thiolated chitosan (chitosan-TBA) coated poly(isobutylcyanoacrylate) (PIBCA) nanoparticles as drug carriers for mucosal administration. Core-corona nanoparticles were obtained by radical emulsion polymerisation of isobutylcyanoacrylate (IBCA) with chitosan of different molecular weights and different proportions of chitosan/chitosan-TBA. In this work, the effect of these nanoparticles on the paracellular permeability of intestinal epithelium was investigated using the Ussing chamber technique, by adding nanoparticle suspensions in the mucosal side of rat intestinal mucosa. Results showed that permeation of the tracer [14C]mannitol and the reduction of transepithelial electrical resistance (TEER) in presence of nanoparticles were more pronounced in those formulations prepared with intermediate amounts of thiolated polymer. This effect was explained thanks to the high diffusion capacity of those nanoparticles through the mucus layer that allowed them to reach the tight junctions in higher extent. It was concluded that, although a first contact between nanoparticles and mucus was a mandatory condition for the development of a permeation enhancement effect, the optimal effect depended on the chitosan/chitosan-TBA balance and the conformational structure of the particles shell. PMID- 18395431 TI - Dissolution rate enhancement of the novel antitumoral beta-lapachone by solvent change precipitation of microparticles. AB - beta-Lapachone [betaLAP] is a novel antitumor drug, which was recently on clinical trials with promising preliminary results. Problems derived from its low water solubility, its instability in solution and its high therapeutic dose constitute some challenges for pharmaceutical researchers. The purpose of the present work is to enhance the limited dissolution rate of betaLAP by the design of particles using a solvent change precipitation process. The procedure induces the spontaneous crystalline growth of the betaLAP in the presence of a stabilizing polymer (Hydroxypropylmethylcellulose) that limits the size of the particles generated. Physicochemical characterization of microparticles and the betaLAP dissolution rate was carried out. The utility of the betaLAP microcrystals in the development of tablets with adequate dissolution properties was also stated. The procedure was optimized in order to obtain stable and homogeneous particles with a small mean particle size (approximately 3 microm) and a narrow particle size distribution. There were no differences between the drying methods evaluated (in an oven and freeze-drying) with regard to particle morphology or dissolution behaviour, which is almost instantaneous. Tablets having suitable mechanical properties were produced by dry granulation prior to compression. The compression process did not compromise betaLAP dissolution characteristics. PMID- 18395432 TI - How do we compare dose to cells in vitro with dose to live animals and humans? Some experiences with inhaled substances. AB - The inhalation route provides closer contact between the ambient environment and living cells than the other major routes-of-entry to the body. In addition, the ambient air transporting exogenous agents to the close proximity of living cells can maintain the reactivity of such agents until they deposit in the extracellular lining layer of the lungs at micrometer distances from the airway epithelium. While toxicity may often occur following the systemic distribution of exogenous substances via the respiratory tract, it is the situation when toxicity occurs at the airway portal-of-entry that pose a particular challenge in the mapping of the dosimetry. In such case, the volume of distribution of solutes before they may enter the airway cells is very small. Therefore exposures of the airway epithelium are more variable and can be much higher than exposures of viable cells of the skin or the gastro-intestinal tract at similar concentrations in the ambient environment. Especially with aerosol exposures, local concentrations around deposited particles can be exceedingly high. To simulate these exposures in a cell culture is a difficult task. However, in order to bridge the in vivo-in vitro gap two methods can be used: (1) Specially designed in vitro systems to better mimic the physiology/morphology of cells residing in the respiratory tract. (2) Mathematical models to analyse the toxicokinetics of the in vitro systems and extrapolate to the corresponding in vivo situation. Both strategies will be exemplified and discussed. PMID- 18395433 TI - Foreword: Congress on Alternative Test Methods in Inhalation Toxicology. PMID- 18395434 TI - Differential control of the PcoI/PcoR quorum-sensing system in Pseudomonas fluorescens 2P24 by sigma factor RpoS and the GacS/GacA two-component regulatory system. AB - A quorum-sensing (QS) locus PcoI/PcoR had been identified previously in the biological control bacterium Pseudomonas fluorescens 2P24. In this paper, we investigated the upstream regulators that influence the transcription of the N acyl homoserine lactone (AHL) biosynthase gene pcoI using a chromosomal pcoIColon, two colonslacZ fusion reporter strain. Stationary sigma factor RpoS was identified as a negative regulator of QS system using a random mini-Tn5 mutant procedure. Furthermore, deletion mutagenesis and complementation experiments demonstrated that the two-component system GacS/GacA positively regulated the QS system by upregulating pcoI transcription. However, compared with the gacA or gacS mutant, introduction of a second mutation of rpoS in the gacA(-) or gacS(-) background did not lead to further change in the transcriptional expression of the pcoI gene or the synthesis of AHL. Our results suggest that in strain 2P24, RpoS could only play its negative regulatory role on the pcoI gene under a functional GacS/GacA system background. PMID- 18395435 TI - Beta-galactosidase monitoring by a biosensor based on Clark electrode: its optimization, characterization and application. AB - beta-Galactosidase is an hydrolase enzyme that catalyzes the hydrolysis of beta galactosides into monosaccharides. Substrates of different beta-galactosidases include ganglioside GM1, lactosylceramides, lactose, and various glycoproteins. A novel aspect of the activity determination of beta-galactosidase was presented. A glucose oxidase biosensor based on Clark electrode was utilized in order to monitor beta-galactosidase. Immobilization of glucose oxidase was made by gelatin and glutaraldehyde as cross-linker. Several parameters such as glucose oxidase activity, gelatin amount, and glutaraldehyde percentage for cross-linking were optimized. The most important parameter, lactose concentration in working buffer was studied in detail. Optimum temperature, thermal stability, optimum pH, buffer system and its concentration effect on the biosensor system, repeatability, reproducibility, and storage and operational stabilities of the biosensor were identified. A linear detection range for beta-galactosidase was observed between 9.4 x 10(-5) and 3.2 x 10(-2)U/ml. Finally, beta-galactosidase activity in artificial intestinal juice was investigated by the biosensor and the results obtained were compared with a reference spectrophotometric method. PMID- 18395436 TI - Glucose sensor based on nano-baskets of tin oxide templated in porous alumina by plasma enhanced CVD. AB - A feasibility study of glucose oxidase (GOx) immobilized tin oxide thin films, consisting of nano-baskets, for glucose sensing is presented. The nano-baskets of SnO(2) were grown on in-house fabricated anodized aluminum oxide pores of approximately 80-nm diameter using plasma enhanced chemical vapor deposition (PECVD) at an RF power of 60W. Hydrated stannic chloride was used as a precursor and O(2) (20 sccm) as a reactant gas. The deposition was carried out from 350 to 450 degrees C at a pressure of 0.2 Torr for 15 min each. Deposition at 450 degrees C resulted in crystalline film with basket-like (nano-sized) structure. GOx was immobilized by physical adsorption (soaking films in GOx solution containing 1000 units for 3h). Increase in film conductivity was noticed after GOx immobilization. The immobilized films were found sensitive to glucose (C(2)H(12)O(6), dextrose) concentration from 10 to 360 mg/dl. Sensitivity increases linearly with glucose concentration. Nano-baskets resulted in higher sensitivity in comparison with other structures. From the elemental analyses of the films after GOx immobilization, GOx was found covalently attached with tin oxide, as evident by N 1s peak in the photoelectron spectra. A possible sensing mechanism is presented and discussed. PMID- 18395437 TI - Personalizing foods: is genotype necessary? AB - The inescapable conclusion of a just a decade of nutrigenomics research must now be brought to practice. Humans differ in their responses to diet and many of these differences are being assigned to genetic polymorphisms. However, differences in the varying responses to diet between humans are not solely because of genetic variation. Lifestage, lifestyle, prior nutritional and physiological variables and even your mother's microflora all influence the differences between humans. The question becomes: are all of these inputs to an individual's health measurable as part of a nutritional phenotype assessment? The answer to this question is increasingly, yes. As variations in humans can be both measured and even more importantly understood, the implications of those measures to dietary guidance become actionable. More accurate assessment of the inputs to human health and the consequences of those inputs measured as accurate proteomic and metabolomic analyses would bring personalized health to practice far faster than waiting for a predictive knowledge of genetic variation. PMID- 18395438 TI - Mortality among Thorotrast-exposed patients and an unexposed comparison group in the German Thorotrast study. AB - Thorotrast was the brand name of a stabilised colloidal solution of thorium dioxide which was used preferentially as an X-ray contrast medium for arteriography between 1930 and 1950. The administration of the medium led to lifelong chronic alpha-particle irradiation by thorium decay products, mainly in the organs of deposition. Several epidemiological follow-up studies were set up after recognition of these side-effects among which the German study was the largest. After an extended follow-up, by 2004 only nine out of 2326 originally exposed subjects were still alive (while 151 of the comparison group, which originally numbered 1890 subjects, survived) and partially more than 70 years observation and chronic exposure time could be studied allowing for further observations to be made about long-term mortality effects of Thorotrast exposure. Median life-expectancy was shortened by 14 years and mortality increased, affecting total mortality SMR=287 for males, SMR=387 for females) as well as cause-specific, especially liver cancer (SMR=16,695 and SMR=12,680, respectively), and the haematopoietic system (SMR=556 and SMR=504, respectively), but not lung cancer. Mortality (total and selected cause-specific) increased with cumulative time since first exposure. PMID- 18395440 TI - Melatonin prevents glucocorticoid inhibition of cell proliferation and toxicity in hippocampal cells by reducing glucocorticoid receptor nuclear translocation. AB - Glucocorticoids are the main product of the adrenal cortex and participate in multiple cell functions as immunosupressors and modulators of neural function. Within the brain, glucocorticoid activity is mediated by high-affinity mineralocorticoid and low-affinity glucocorticoid receptors. Among brain cells, hippocampal cells are rich in glucocorticoid receptors where they regulate excitability and morphology. Also, elevated glucocorticoid levels suppress hippocampal neurogenesis in adults. The pineal neuroindole, melatonin, reduces the affinity of glucocorticoid receptors in rat brain and prevents glucocorticoid induced apoptosis. Here, the ability of melatonin to prevent glucocorticoid induced cell death in hippocampal HT22 cells was investigated in the presence of neurotoxins. Results showed that glucocorticoids reduce cellular growth and also enhance sensitivity to neurotoxins. We found a G(1) cell cycle arrest mediated by an increase of cyclin/cyclin-dependent kinase inhibitor p21(WAF1/CIP1) protein after dexamethasone treatment and incremental change in amyloid beta protein and glutamate toxicity. Melatonin prevents glucocorticoids inhibition of cell proliferation and reduces the toxicity caused by glucocorticoids when cells were treated with dexamethasone in combination with neurotoxins. Although, melatonin does not reduce glucocorticoid receptor mRNA or protein levels, it decreases receptor translocation to nuclei in these cells. PMID- 18395439 TI - Ligand specificity and evolution of liver X receptors. AB - Liver X receptors (LXRs) are key regulators of lipid and cholesterol metabolism in mammals. Little is known, however, about the function and evolution of LXRs in non-mammalian species. The present study reports the cloning of LXRs from African clawed frog (Xenopus laevis), Western clawed frog (Xenopus tropicalis), and zebrafish (Danio rerio), and their functional characterization and comparison with human and mouse LXRs. Additionally, an ortholog of LXR in the chordate invertebrate Ciona intestinalis was cloned and functionally characterized. Ligand specificities of the frog and zebrafish LXRs were very similar to LXRalpha and LXRbeta from human and mouse. All vertebrate LXRs studied were activated robustly by the synthetic ligands T-0901317 and GW3965 and by a variety of oxysterols. In contrast, Ciona LXR was not activated by T-0901317 or GW3965 but was activated by a limited number of oxysterols, as well as some androstane and pregnane steroids. Pharmacophore analysis, homology modeling, and docking studies of Ciona LXR predict a receptor with a more restricted ligand-binding pocket and less intrinsic disorder in the ligand-binding domain compared to vertebrate LXRs. The results suggest that LXRs have a long evolutionary history, with vertebrate LXRs diverging from invertebrate LXRs in ligand specificity. PMID- 18395441 TI - Structure-activity relationships of synthetic progestins in a yeast-based in vitro androgen bioassay. AB - The recent identification of tetrahydrogestrinone (THG), a non-marketed designer androgen used for sports doping but previously undetectable by established mass spectrometry-based urine drug screens, and its production by a facile chemical modification of gestrinone has raised concerns about the risks of developing designer androgens from numerous marketed progestins. We therefore have used yeast-based in vitro androgen and progesterone bioassays to conduct a structure activity study assessing the intrinsic androgenic potential of commercially available progestins and their derivatives, to identify those compounds or structures with the highest risk of forming a basis for such misapplication. Progestins had a wide range of androgenic bioactivity that was not reliably predicted for individual steroids by their progestin bioactivity. 17alpha Hydroxyprogesterone and 19-norprogesterone derivatives with their bulky 17beta substituents were strong progestins but generally weak androgens. 17alpha Ethynylated derivatives of testosterone, 19-nortestosterone and 18-methyl-19 nortestosterone such as gestrinone, ethisterone, norethisterone and norgestrel had the most significant intrinsic androgenicity of all the commercially marketed progestins. Facile chemical modification of the 17alpha-ethynyl group of each of these progestins produces 17alpha-methyl, ethyl and allyl derivatives, including THG and norbolethone, which further enhanced androgenic bioactivity. Thus by using the rapid and sensitive yeast bioassay we have screened a comprehensive set of progestins and associated structures and identified the ethynylated testosterone, 19-nortestosterone and 18-methyl-19-nortestosterone derivatives as possessing the highest risk for abuse and potential for conversion to still more potent androgens. By contrast, modern progestins such as progesterone, 17alpha hydroxyprogesterone and 19-norprogesterone derivatives had minimal androgenic bioactivity and pose low risk. PMID- 18395442 TI - Diphenidol-related diamines as novel muscarinic M4 receptor antagonists. AB - A series of hydrochloride derivatives 2a-9a and quaternary ammonium derivatives 3b-9b of diphenidol have been synthesized and characterized in receptor binding and cellular functional assays versus human muscarinic M(1)-M(5) receptors expressed in CHO cells. Compound 8b, a methiodide derivative with a bipiperidinyl moiety and a second diphenidol framework, showed a potent and selective M(4) activity as competitive antagonist. Moreover 8b, acting as an allosteric modulator, was able to retard the dissociation rate of [(3)H]-N-methylscopolamine from CHO-M(4) cell membranes exposed to atropine. Taken together, these data suggest that 8b might open new avenues to the discovery of novel multivalent antagonists for the muscarinic receptors. PMID- 18395443 TI - Discovery and preclinical studies of 5-isopropyl-6-(5-methyl-1,3,4-oxadiazol-2 yl)-N-(2-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)pyrrolo[2,1-f][1,2,4]triazin-4 amine (BMS-645737), an in vivo active potent VEGFR-2 inhibitor. AB - We report herein a series of substituted N-(1H-pyrrolo[2,3-b]pyridin-5 yl)pyrrolo[2,1-f][1,2,4]triazin-4-amines as inhibitors of vascular endothelial growth factor receptor-2 tyrosine kinase. Through structure-activity relationship studies, biochemical potency, pharmacokinetics, and kinase selectivity were optimized to afford BMS-645737 (13), a compound with good preclinical in vivo activity against human tumor xenograft models. PMID- 18395444 TI - A novel pyrene-guanidiniocarbonyl-pyrrole cation efficiently differentiates between ds-DNA and ds-RNA by two independent, sensitive spectroscopic methods. AB - At micromolar concentrations and equimolar conditions in respect to basepairs, a novel pyrene-guanidiniocarbonyl-pyrrole cation 1 exhibited a strong ICD signal at about lambda=300nm specifically upon the interaction with ds-DNA, while under the same conditions a new fluorescence maximum at lambda=480nm appeared exclusively upon the addition of ds-RNA. PMID- 18395445 TI - Candidate screening of the bovine and feline spinal muscular atrophy genes reveals no evidence for involvement in human motor neuron disorders. AB - Spinal muscular atrophy (SMA) is a devastating neuromuscular disease characterised by progressive loss of spinal motor neurons. Mutations in the genes underlying spontaneous bovine and feline models of SMA have recently been described. The clinical and pathological features of these disorders are similar to human forms of SMA making both genes excellent candidates in patients with motor neuron loss of no known aetiology. Here we report that a screen for mutations in coding regions and splice sites of the LIX1 and FVT1 genes in a cohort of 96 non-5q SMA patients and 119 familial and sporadic Amyotrophic Lateral Sclerosis patients identified no obvious pathogenic changes. This study indicates that mutations in these genes do not contribute significantly to the cause of motor neuron diseases in the human population. PMID- 18395446 TI - Modulation of mitochondrial morphology by bioenergetics defects in primary human fibroblasts. AB - Mitochondria are dynamic organelles with continuous fusion and fission, the equilibrium of which results in mitochondrial morphology. Evidence points to there being an intricate relationship between mitochondrial dynamics and oxidative phosphorylation. We investigated the bioenergetics modulation of mitochondrial morphology in five control cultured primary skin fibroblasts and seven with genetic alterations of oxidative phosphorylation. Under basal conditions, control fibroblasts had essentially filamentous mitochondria. Oxidative phosphorylation inhibition with drugs targeting complex I, III, IV or V induced partial but significant mitochondrial fragmentation, whereas dissipation of mitochondrial membrane potential (D Psi m) provoked complete fragmentation, and glycolysis inhibition had no effect. Oxidative phosphorylation defective fibroblasts had essentially normal filamentous mitochondria under basal conditions, although when challenged some of them presented with mild alteration of fission or fusion efficacy. Severely defective cells disclosed complete mitochondrial fragmentation under glycolysis inhibition. In conclusion, mitochondrial morphology is modulated by D Psi m but loosely linked to mitochondrial oxidative phosphorylation. Its alteration by glycolysis inhibition points to a severe oxidative phosphorylation defect. PMID- 18395447 TI - A review of equine muscle disorders. AB - Muscle disorders are a common cause of disability in horses. For many years, clinical manifestations such as muscle pain, exercise intolerance, weakness, and stiffness were believed to be caused by a single syndrome. However, in the past years a broad spectrum of muscle disorders have been recognized including glycogen and polysaccharide storage myopathies, malignant hyperthermia, mitochondrial myopathy, hyperkalemic periodic paralysis and others. For some, a specific mutation has been identified. Recognition of the myopathic clinical phenotype and thorough clinical, electrodiagnostic, and histological evaluations are essential to further our understanding of equine myopathies. Advances in understanding equine myopathies may potentially benefit other species including humans. PMID- 18395448 TI - Left ventricular non-compaction in a patient with myotonic dystrophy type 2. AB - Cardiac involvement is frequent in myotonic dystrophy type 2 (DM2) with dilated cardiomyopathy and severe arrhythmias having been reported. Left ventricular non compaction is a cardiomyopathy often associated with neuromuscular disorders. We report the case of a 61-year-old man with DM2 treated for 5 years for a suspected dilated cardiomyopathy. Echocardiography showed left ventricular non-compaction typical pattern, with prominent apical trabeculations and intertrabecular spaces perfused from ventricular cavity. MRI confirmed the diagnosis. Physicians should be aware of the risk of severe cardiac complications in DM2 patients. Left ventricular non-compaction diagnosis is often overlooked. Neurological examination should be performed in all patients with left ventricular non compaction. PMID- 18395449 TI - Similarities and differences in aneurysmal subarachnoid haemorrhage between eastern Finland and northern Sydney. AB - A significant body of literature on aneurysmal subarachnoid haemorrhage has emerged from Finland. The Finnish source from a small founding population, rather than from a wide heterogeneous source such as used for other populations, suggests the need for caution when attempting to generalise using Finnish data. This study aims to identify the differences and similarities between the aneurysmal subarachnoid haemorrhage populations of eastern Finland and northern Sydney to ascertain whether information that is derived from Finland has applicability to an Australian context. Existing aneurysmal subarachnoid haemorrhage databases of Kuopio University Hospital in eastern Finland and the Royal North Shore and Dalcross Private Hospitals in northern Sydney from 2000 to 2005 were combined and analysed. A total of 879 patients were identified. Comparisons fell into three categories: features that were similar between the two populations; those with apparent differences that may be explained by methods of data collection or referral patterns; and differences that probably represent a true difference between these populations. The differences suggest that genetic predispositions for aneurysm development may not affect the likelihood for aneurysmal rupture. PMID- 18395450 TI - Subependymal giant cell astrocytoma with intratumoral hemorrhage in the absence of tuberous sclerosis. AB - Subependymal giant cell astrocytoma (SEGA) is an uncommon tumor that usually occurs in the setting of tuberous sclerosis (TS) syndrome. We report a rare case of an intratumoral and a small intraventricular hemorrhage complicating a SEGA in an adult patient without any signs of TS. Although pre-operative CT and MRI findings for the tumor were typical of SEGA, SEGA was not considered in the differential diagnosis because the patient was lacking any symptoms of TS. This is the second report of intraventricular and intratumoral hemorrhage complicating a SEGA and the first case in which these complications occurred in an adult patient in whom there was no previous suspicion of systemic disease. PMID- 18395451 TI - Muscle atrophy associated with multiple sclerosis: a benign condition or the onset of amyotrophic lateral sclerosis? AB - When muscular atrophy develops in multiple sclerosis (MS) patients, its etiology may vary from benign to serious. We describe an individual with a 24-year history of MS who developed amyotrophic lateral sclerosis (ALS). The literature is reviewed with particular attention to the clinical and electrophysiologic characteristics of patients with MS and muscular atrophy and to those rare patients with concurrent MS and ALS. Circumstances warranting a heightened suspicion for concurrent ALS are discussed and careful evaluation of similar patients is encouraged. PMID- 18395452 TI - Traumatic intracranial aneurysm: a brief review. AB - Traumatic intracranial aneurysms are uncommon and represent fewer than 1% of all cerebral aneurysms. They may develop after blunt or penetrating head injuries and can present both diagnostic challenges and surgical difficulties. Because traumatic aneurysms are fragile and prone to rupture, early diagnosis with cerebral angiography and prompt treatment are essential. We present two patients with traumatic aneurysms and discuss their aetiology, classification, clinical presentations, diagnosis, and treatment options. PMID- 18395453 TI - Venous and arterial thrombo-embolic complications of hormonal treatment in a male to-female transgender patient. AB - We present a male-to-female (MTF) transgender patient admitted with a pulmonary embolism. The patient had been treated with high-dose oestrogens since the age of 16. Following a prolonged period of hypotension, our patient sustained cerebral border zone infarcts. There was evidence of bilateral carotid stenosis on Doppler ultrasound. We discuss the treatment and vascular complications of gender dysphoria. PMID- 18395454 TI - Carboxymethylated pyridoindole antioxidants as aldose reductase inhibitors: Synthesis, activity, partitioning, and molecular modeling. AB - Starting from the efficient hexahydropyridoindole antioxidant stobadine, a series of carboxymethylated tetrahydro- and hexahydropyridoindole derivatives was synthesized and tested for the inhibition of aldose reductase, an enzyme involved in the etiology of diabetic complications. In vitro inhibiton of rat lens aldose reductase was determined by a conventional method. Kinetic analysis of (2-benzyl 2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-8-yl)-acetic acid (5b) and (2-phenethyl 2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-8-yl)-acetic acid (5c), the most potent compounds in this series with activities in micromolar range, showed uncompetitive inhibition. In addition to the importance of the acidic function, the inhibition efficacy was highly influenced by the steric conformation of the lipophilic aromatic backbone when comparing tetrahydro- and hexahydropyridoindole congeners. Selectivity with respect to the closely related aldehyde reductase was determined by measuring the corresponding inhibitory activities. Antioxidant action of the novel compounds was documented in a DPPH test and in a liposomal membrane model, oxidatively stressed by peroxyl radicals. The presence of a basicity center at the tertiary nitrogen, in addition to the acidic carboxylic function, predisposes these compounds to form double charged zwitterionic species, a characteristic which may remarkably affect their pH-lipophilicity profile. For compounds 5b and 5c, a maximal distribution ratio in a system comprised of 1-octanol/phosphate buffer was recorded near the neutral physiological pH, the region where the isoelectric point lies. Molecular docking simulations into the ALR2 active site performed for the zwitterionic species provided an explanation for the observed structure-activity relationships and the calculated parameters were in agreement with characteristic differences in the stereoelectronic profiles of the tetrahydro- versus hexahydropyridoindoles. 'Drug likeness' of the novel aldose reductase inhibitors was assessed by applying the criteria of Lipinski's 'rule of five'. PMID- 18395455 TI - Additive SMILES-based optimal descriptors in QSAR modelling bee toxicity: Using rare SMILES attributes to define the applicability domain. AB - The additive SMILES-based optimal descriptors have been used for modelling the bee toxicity. The influence of relative prevalence of the SMILES attributes in a training and test sets to the models for bee toxicity has been analysed. Avoiding the use of rare attributes improves statistical characteristics of the model on the external test set. The possibility of using the probability of the presence of SMILES attributes in training and test sets for rational definition of the applicability domain is discussed. PMID- 18395456 TI - Influence of the Si cap layer on the SiGe islands morphology. AB - Transmission electron microscopy (TEM), atomic force microscopy (AFM), and EDX methods were used to study morphology and chemical composition of SiGe/Si(001) islands grown at 700 degrees C and covered at 550 degrees C and 700 degrees C by Si layers of different thickness. The samples were grown in ultra high vacuum chemical vapor deposition process (UHVCVD) controlled with in situ reflection of high-energy electron diffraction (RHEED). The islands transformed from initial pyramid and dome shapes to lens shape for 1.4 nm and 3.7 nm cap layer thickness at 550 degrees C and 700 degrees C, respectively. An increase of lateral to vertical ratio was observed during the transformation. For the higher depositing temperature the ratio was bigger and was increasing continuously with cap layer thickness. Also, with increasing Si cap layer thickness, the Ge concentration was decreasing, which was more observable for higher capping temperature. PMID- 18395457 TI - Optical properties and bandgaps from low loss EELS: pitfalls and solutions. AB - We investigate the impacts of zero loss peak (ZLP) removal and retardation effects altering the electron energy loss spectrum on the optical properties obtained by using Kramers-Kronig analysis and on the determination of the bandgap. For this purpose we use amorphous SiN(x):H having a bandgap of Eg(SiNx:H)= 5.5 eV. We demonstrate that for bandgap determination not only the accurate removal of the ZLP is crucial, moreover also retardation losses have to be taken into account. The same is valid for an accurate determination of the optical properties of semiconductors which can be done if the retardation effects are treated correctly or avoided at all before Kramers-Kronig analysis is applied. Beside the detailed study on using SiN(x):H we discuss the impact of the retardation effects on several other semiconductors and insulators, like GaP. PMID- 18395458 TI - Application of "wet" extrapolation method for activity standardisation of electron capture radionuclides. AB - The activity of electron capture radionuclides is usually determined by 4pi(proportional counter, PC)-gamma coincidence counting. The corrections necessary for the final activity value calculation are obtained by an extrapolation method. Variation of the PC detection efficiency can be achieved through different methods, e.g. by changing the self-absorption of the source using absorbing foils or by adding carriers. Another possibility is a "wet" extrapolation method, which utilises an absorption change during the drying of a water droplet added onto the source surface. In this paper, slopes of extrapolation curves and resulting activity values obtained by different methods are compared for several radionuclides ((54)Mn, (139)Ce, (88)Y, (57)Co). In some cases a digital coincidence system was used for the analysis of measured data. The "wet" extrapolation, due to its very simple procedure, seems to be convenient for routine measurement and its accuracy is similar to the other methods. PMID- 18395459 TI - Tiagabine is neuroprotective in the N171-82Q and R6/2 mouse models of Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by chorea, incoordination, and shortened life-span, and by huntingtin inclusions and neurodegeneration. We previously screened the 1040 FDA approved compounds from the NINDS compound library and found that a compound, nipecotic acid, significantly reduced mutant huntingtin aggregations and blocked cell toxicity in an inducible cell model of HD. Because nipecotic acid does not cross the blood-brain barrier (BBB), we studied its analogue, tiagabine, which is able to cross the BBB, in both N171-82Q and R6/2 transgenic mouse models of HD. Tiagabine was administered intraperitoneally at 2 and 5 mg/kg daily in HD mice. We found that tiagabine extended survival, improved motor performance, and attenuated brain atrophy and neurodegeneration in N171-82Q HD mice. These beneficial effects were further confirmed in R6/2 HD mice. The levels of tiagabine at effective doses in mouse serum are comparable to the levels in human patients treated with tiagabine. These results suggest that tiagabine may have beneficial effects in the treatment of HD. Because tiagabine is an FDA-approved drug, it may be a promising candidate for future clinical trials for the treatment of HD. PMID- 18395460 TI - Activation of brain metabolism and fos during limbic seizures: the role of locus coeruleus. AB - The noradrenergic nucleus Locus Coeruleus (LC) densely innervates limbic structures. In rats, the damage to LC by the neurotoxin DSP-4, converts episodic limbic seizures induced by bicuculline infusion in the anterior piriform cortex (APC) into self-sustaining status epilepticus (SE). SE induced by this approach is similar to SE induced by co-infusing cyclothiazide and bicuculline into APC in rats bearing an intact LC. As opposed to other commonly used rat SE models (e.g. systemic kainate or pilocarpine), this approach allows one to analyze the effects of SE on brain regions which are solely due to spreading of seizure activity, rather than to direct effect of systemic chemoconvulsant. We evaluated the expression of Fos protein (an immediate early gene product), and the local cerebral metabolic rates for [14C] 2-deoxyglucose (lCMRglc), in rats following SE induced either by cyclothiazide+bicuculline or by DSP-4+bicuculline. We demonstrated that regional Fos expression after SE does not parallel the increase in lCMRglc, in LC-lesioned rats. In DSP-4+bicuculline rats there is an overall lower expression of the protein as compared with the cyclothiazide+bicuculline or bicuculline alone groups; even more, such a difference co-exists with an higher lCMRglc in the DSP-4+bicuculline-treated rats in some regions, as compared with the other groups. These data show that LC neurons play an important role in determining immediate early genes expression even in conditions of strong pathological activation, such as limbic SE. This might have relevant effects in the plastic mechanisms related with epileptogenesis. PMID- 18395461 TI - Revised determination of free and complexed myrosinase activities in plant extracts. AB - The enzyme myrosinase (thioglucoside glucohydrolase, EC 3.2.1.147, formerly EC 3.2.3.1) catalyzes the hydrolysis of glucosinolates after tissue damage in plants of the order Brassicales. The various myrosinase isoforms occur either as free soluble dimers or as insoluble complexes. We propose a reliable method for determination of both soluble and insoluble myrosinase activity concentrations in partially purified plant extracts. The procedure requires the removal of endogenous glucosinolates through ion-exchange columns previous to enzyme measurements. Myrosinase activity was assayed in continuous mode by photometric quantification of the released glucose using glucose-oxidase with peroxidase and colorimetric indicators. The measurement of the colored product at 492nm has a favorable signal to noise ratio both in clear extract solutions (free dimers) and in turbid pellet suspensions (insoluble complexes). No interferences by ascorbic acid were found in continuous analyses. With the recommended sample preparation methods and assay conditions potential activities in damaged plant tissues can be characterized which are involved in plant defense mechanisms. PMID- 18395462 TI - About human tumor antigens to be used in immunotherapy. AB - The choice of antigens to be used in cancer immunotherapy remains a crucial and difficult issue. This review highlights some properties of the different groups of human tumor antigens recognized by T lymphocytes, focusing on parameters that should influence this choice, such as tumor specificity and level of antigen expression. PMID- 18395463 TI - The fast kinetics of AMPA GluR3 receptors is selectively modulated by the TARPs gamma 4 and gamma 8. AB - Transmembrane AMPA receptor-associated regulatory proteins (TARPs) modulate the kinetics of AMPA receptors and increase their surface expression. This study compared the effects of the TARPs gamma2, gamma3, gamma4 and gamma8 on the AMPA receptor subunits GluR1-3. We found that the deactivation and desensitization time constants of GluR3 receptors were greatly increased by gamma4 and gamma8 (approximately 3 fold) whereas gamma2 and gamma3 caused minimal changes. This stands in contrast to the effects on GluR1 and GluR2 receptors which were similar across all four TARPs. Other response parameters like the current density, the EC(50) for glutamate-induced peak currents, and kainate-induced currents were in general more effectively modulated by gamma2 and gamma3 in all subunits, including GluR3. All TARPs were effective in increasing the unitary conductance. The differential TARP effects on deactivation time constants of the GluR3 subunit are likely to have a significant impact on synaptic responses across different neurons in the brain. PMID- 18395464 TI - Timing and duration of incarceration and high-risk sexual partnerships among African Americans in North Carolina. AB - PURPOSE: Incarceration may contribute to HIV transmission by disrupting stable partnerships and promoting high-risk partnerships. We investigated incarceration and high-risk partnerships among African Americans in North Carolina. METHODS: We conducted a weighted analysis using the North Carolina Rural Health Project (N = 320), a population-based case-control study of HIV among African Americans. We measured associations between timing and duration of incarceration and high-risk partnerships (multiple partnerships or sex trade for money or drugs). RESULTS: Duration of incarceration appeared to be more important than how long ago incarceration occurred. After adjustment for sociodemographic indicators, high risk partnerships were associated with short-term (<1 month) incarceration of the respondent versus no respondent incarceration (men: adjusted prevalence ratio (aPR) 1.9, 95% confidence interval (95% CI) 1.2-2.8; women: aPR 3.1, 95% CI 1.2 8.3). High-risk partnerships also were associated with incarceration of a partner versus no partner incarceration (men: aPR 1.8, 95% CI 1.1-3.0; women: aPR 2.0, 95% CI 1.1-3.8). Among men, associations remained when adjusting for substance use. Among women, adjustment for substance use weakened estimates due to the strong correlation between substance use and incarceration. CONCLUSIONS: HIV prevention programs targeting currently and formerly incarcerated individuals and their partners may decrease HIV in African American communities with high incarceration rates. PMID- 18395465 TI - Historical records as a source of information for childhood socioeconomic status: results from a pilot study of decedents. AB - PURPOSE: The validity of parental occupation recalled by adult children is not established, yet it is commonly used to measure childhood socioeconomic status (SES). We investigated the feasibility of using data from historical records to validate recalled parental SES. METHODS: Data from death certificates and applications for Social Security numbers (parents' names, date and place of birth) were used to locate birth certificates and 1930 census records of 416 decedents in Forsyth County, NC, to verify parental occupation and childhood residence. RESULTS: Birth certificates and/or census records were located for 85% of decedents. Of 257 for whom both records were searched, both were found for 60%, only a census record for 10%, and only a birth certificate for 24%. Among those with father's occupation recorded on both records (n = 138), occupational category matched on 89% of records (kappa = 0.86). Place of residence/birth, which can be linked with census-based county socioeconomic indicators, was also highly concordant across records. CONCLUSIONS: These results demonstrate that birth and census records can be located for most decedents and that the childhood SES data contained therein is highly concordant. Thus they are an alternative to recalled childhood SES and a source of validation data in life course studies. PMID- 18395466 TI - The effect of forearm support on children's head, neck and upper limb posture and muscle activity during computer use. AB - Use of computers by children has increased rapidly, however few studies have addressed factors which may reduce musculoskeletal stress during computer use by children. This study quantified the postural and muscle activity effects of providing forearm support when children used computers. Twelve male and 12 female children (10-12 years) who regularly used computers were recruited. Activities were completed using a computer with two workstation configurations, one of which provided for forearm support on the desk surface. 3D posture was analysed using an infra-red motion analysis system. Surface EMG was collected from five muscle groups in the neck/shoulder region and right upper limb. Providing a support surface resulted in more elevated and flexed upper limbs. The use of forearm or wrist support was associated with reduced muscle activity for most muscle groups. Muscle activity reductions with support were of sufficient magnitude to be clinically meaningful. The provision of a supporting surface for the arm is therefore likely to be useful for reducing musculoskeletal stresses associated with computing tasks for children. PMID- 18395467 TI - Self-memory biases in explicit and incidental encoding of trait adjectives. AB - An extensive literature has demonstrated that encoding information in a self referential manner enhances subsequent memory performance. This 'self-reference effect' (i.e., better memory for self-referent than other-referent information) is generally elicited in paradigms that require participants to evaluate the self descriptiveness of personality characteristics. Extending work of this kind, the current research explored the possibility that explicit evaluative processing is not a necessary precondition for the emergence of this effect. Rather, responses to self cues may enhance item encoding even in the absence of explicit evaluative instructions. We explored this hypothesis by testing memory for items encoded in either an evaluative or relational context. The relational context was achieved by requesting participants to report the spatial relationship between target stimuli, and visual or verbal referent cues. The results revealed a self-referent memory advantage, regardless of the encoding context or triggering cue. The theoretical implications of these findings are considered. PMID- 18395468 TI - Increased cortico-cortical functional connectivity in early-stage Parkinson's disease: an MEG study. AB - We set out to determine whether changes in resting-state cortico-cortical functional connectivity are a feature of early-stage Parkinson's disease (PD), explore how functional coupling might evolve over the course of the disease and establish its relationship with clinical deficits. Whole-head magnetoencephalography was performed in an eyes-closed resting-state condition in 70 PD patients with varying disease duration (including 18 recently diagnosed, drug-naive patients) in an "OFF" medication state and 21 controls. Neuropsychological testing was performed in all subjects. Data analysis involved calculation of three synchronization likelihood (SL, a general measure of linear and non-linear temporal correlations between time series) measures which reflect functional connectivity within (local) and between (intrahemispheric and interhemispheric) ten major cortical regions in five frequency bands. Recently diagnosed, drug-naive patients showed an overall increase in alpha1 SL relative to controls. Cross-sectional analysis in all patients revealed that disease duration was positively associated with alpha2 and beta SL measures, while severity of parkinsonism was positively associated with theta and beta SL measures. Moderately advanced patients had increases in theta, alpha1, alpha2 and beta SL, particularly with regard to local SL. In recently diagnosed patients, cognitive perseveration was associated with increased interhemispheric alpha1 SL. Increased resting-state cortico-cortical functional connectivity in the 8-10 Hz alpha range is a feature of PD from the earliest clinical stages onward. With disease progression, neighboring frequency bands become increasingly involved. These findings suggest that changes in functional coupling over the course of PD may be linked to the topographical progression of pathology over the brain. PMID- 18395469 TI - Individual alpha-frequency correlates with amplitude of visual evoked potential and hemodynamic response. AB - In a simultaneous electroencephalography (EEG) and near-infrared spectroscopy (NIRS) study, the predictive value of the individual alpha-frequency at rest (IAF) for the amplitude of neuronal and vascular responses to visual stimulation was investigated. Across subjects, we find (i) an inverse relationship between IAF and the amplitude of the alpha-rhythm at rest. The IAF also predicts (ii) the amplitude of the visual evoked potential (VEP), as well as (iii) the amplitude of the alpha-rhythm during stimulation. Most importantly, (iv) IAF correlates with the oxygenation response to visual stimulation: A high IAF predicts a low alpha amplitude at rest, a small VEP amplitude and a small oxygenation response. Conversely, a low IAF predicts high alpha-amplitude and larger electrophysiological and vascular responses to stimulation. Based on these findings, we assume that the relationship between IAF and neuronal and vascular response stems from the size of the network recruited for visual processing. The relation between IAF, alpha-amplitude, evoked potential and vascular response is discussed in the framework of a simple heuristic model. The results may partly explain the large intersubject variability observed in recently published concurrent EEG-fMRI studies. PMID- 18395470 TI - Neural correlates of age-related visual search decline: a combined ERP and sLORETA study. AB - Differences in the neural systems underlying visual search processes for young (n=17, mean age 19.6+/-1.9) and older (n=22, mean age 68.5+/-6) subjects were investigated combining the Event-Related Potential (ERP) technique with standardized Low-Resolution brain Electromagnetic Tomography (sLORETA) analyses. Behavioral results showed an increase in mean reaction times (RTs) and a reduction in hit rates with age. The ERPs were significantly different between young and older subjects at the P3 component, showing longer latencies and lower amplitudes in older subjects. These ERP results suggest an age-related decline in the intensity and speed of visual processing during visual search that imply a reduction in attentional resources with normal aging. The sLORETA data revealed a significantly reduced neural differentiation in older subjects, who recruited bilateral prefrontal regions in a nonselective manner for the different search arrays. Finally, sLORETA between-group comparisons revealed that relative to young subjects, older subjects showed significantly reduced activity in anterior cingulate cortex as well as in numerous limbic and occipitotemporal regions contributing to visual search processes. These findings provide evidence that the neural circuit supporting this cognitive process is vulnerable to normal aging. All these attentional factors could contribute to poorer performance of older compared to young subjects in visual search tasks. PMID- 18395471 TI - Diffusion tensor imaging of normal white matter maturation from late childhood to young adulthood: voxel-wise evaluation of mean diffusivity, fractional anisotropy, radial and axial diffusivities, and correlation with reading development. AB - Using diffusion tensor MR imaging (DTI) and advanced voxel-wise analysis tools, we study diffusivity and anisotropy changes of white matter from late childhood to young adulthood, and correlate quantitative diffusion indices with Chinese and English reading performance scores. Seventy-five normal healthy school going ethnic Chinese students and young adults of three age groups were recruited (group 1, n=24, mean+/-SD=7.4+/-0.3 years; group 2, n=27, mean+/-SD=10.3+/-0.5 years; group 3, n=24, mean+/-SD=22.8+/-2.3 years). DTI was performed with 3 mm isotropic resolution to cover the entire brain. Voxel-wise analysis was performed using Tract-Based Spatial Statistics (TBSS) to localize regions of white matter showing significant changes of fractional anisotropy (FA), mean diffusivity (MD), and axial and radial diffusivities between groups. We found increased FA and decreased MD with increasing age in regions of cerebellar white matter, right temporal white matter, and a large portion of the superior frontal and parietal white matter driven by both the reduction of radial diffusivity and axial diffusivity with the former to a greater extent. Changes were continual from late childhood to young adulthood. Findings were confirmed by region-of-interest analysis in specific white matter tracts. After controlling for the effect of age, significant correlation was found between diffusion indices of the anterior limb of the left internal capsule and Chinese reading score (p=0.05), and of the corona radiata and English reading score (p=0.026 and p=0.029 for left and right, respectively). These DTI indices likely reflect the multiple biological processes that occur during brain development which provide the neural substrate for ongoing functional connectivity including for reading development. PMID- 18395472 TI - Is retaining the youthful functional anatomy underlying speed of information processing a signature of successful cognitive ageing? An event-related fMRI study of inspection time performance. AB - It has been hypothesized that individual differences in cognitive ageing might in part be based on the relative preservation of speed of information processing. However, the biological foundations of processing speed are not understood. Here we compared two groups of non-demented older people who had relatively similar IQs at age 11 but differed markedly in non-verbal reasoning ability at age 70: 'cognitive sustainers' (n=25), and 'cognitive decliners' (n=15). Using an event related fMRI design, we studied the BOLD response while they performed an inspection time task. Inspection time is a two-alternative forced choice, backward masking test of the speed of the early stages of visual information processing. Inspection time has a well-established, significant association with higher cognitive abilities. The group of cognitive sustainers showed a pattern of BOLD activation-deactivation in response to inspection time stimulus duration differences that was similar to a healthy young sample [Deary, I.J., Simonotto, E., Meyer, M., Marshall, A., Marshall, I., Goddard, N., Watdlaw, J.M., 2004a. The functional anatomy of inspection time: an event-related fMRI study. NeuroImage 22, 1466-1479]. The group of cognitive decliners lacked these clear neural networks. The relative preservation of complex reasoning skills in old age may be associated with the preservation of the neural networks that underpin fundamental information processing in youth. PMID- 18395473 TI - Psychological treatment of patients with psychogenic non-epileptic seizures: an outcome study. AB - INTRODUCTION: It is estimated that up to 25% of patients referred to specialised epilepsy centers suffer from psychogenic non-epileptic seizures (PNES). The prognosis is unfavourable and there are no generally accepted treatment protocols. METHOD: In this study, the effect of an uncontrolled, prospective inpatient treatment program for PNES patients is evaluated. The treatment is multidisciplinary and based on cognitive behavioural principles. Seizure control, general psychopathology, anxiety, depression, coping, dissociation and health related quality of life are evaluated. Twenty-two patients participated in the study of which 16 patients were followed 6 months after treatment. RESULTS: After follow-up, 81% of patients had a seizure reduction of over 50%, and half of them became seizure-free. Measures of anxiety, depression and dissociation tended to normalize, coping was more adequate and health related quality of life was increased slightly. In the period between the end of treatment and follow-up the most positive effects are maintained and even strengthened. Patients who became seizure-free at follow-up improved more on the psychological outcome measures than patients with continuing seizures. CONCLUSION: The outcome suggests effectiveness of the treatment. PNES patients may profit from a comprehensive, multidisciplinary treatment program following cognitive behavioural principles. Seizure cessation appears to be an important factor in the improvement of psychological functioning. PMID- 18395474 TI - Acetaminophen and the sodium balance during febrile seizures. PMID- 18395476 TI - Time course and efficiency of protein synthesis inhibition following intracerebral and systemic anisomycin treatment. AB - Since its discovery in the 1960s, anisomycin has been used for studying the impact of protein synthesis for manifold cerebral processes such as long-term plastic changes after learning. The common limitation of nearly all pharmacological experiments, including anisomycin treatment, is to precisely verify the affected brain regions. Here we illustrate anisomycin effects on protein synthesis in distinct brain regions of mice (C57BL/6JOlaHsd), revealing differences between three modes of anisomycin application (subcutaneous, s.c.; intraperitoneal, i.p.; local microinfusions into the hippocampus). Our method is based on inhibition of the incorporation of the radioactively-labelled amino acids [(35)S]-Methionine/Cysteine into newly synthesised proteins. Washing the brain slices before autoradiography removes pools of amino acids, which have not been incorporated into newly synthesised proteins, thus, illustrating pure protein synthesis. By comparing different routes of systemic anisomycin application (i.p. versus s.c.; 150 mg/kg) it became evident that the effect of i.p. injection of anisomycin is fully reversed after 6 h, whereas s.c. injection is inhibiting protein synthesis in the hippocampus even 9 h after application. Local microinfusions of anisomycin into the hippocampus were shown to have long lasting effects as well, which reversed as late as 9 h after injection. The diffusion of anisomycin was maximal at 3 h after injection and more precisely confined to the intended area using a lower dose (20 microg/site) instead of the commonly used dose of 62.5 microg/site. The broad time window of anisomycin action, as revealed in our study, has to be considered, if it comes to the interpretation of time course studies within the context of protein synthesis dependent processes. PMID- 18395475 TI - Effects of sustained interstitial fluid pressurization under migrating contact area, and boundary lubrication by synovial fluid, on cartilage friction. AB - OBJECTIVE: This experimental study tests two hypotheses which address outstanding questions in cartilage lubrication: can the friction coefficient remain low under sustained physiological loading conditions? How effective is synovial fluid (SF) in the lubrication of articular cartilage? Based on theory, it is hypothesized that migrating contact areas can maintain elevated cartilage interstitial fluid pressurization, thus a low friction coefficient, indefinitely. It is also hypothesized that the beneficial effects of SF stem from boundary lubrication rather than fluid-film lubrication. DESIGN: Five experiments were conducted on immature bovine femoro-tibial joints, to compare the frictional response under migrating vs stationary contact areas; the frictional response in SF vs saline; the role of sliding velocity and the role of congruence on the friction coefficient. RESULTS: Migrating contact area could maintain a low friction coefficient under sustained physiological conditions of loading for at least 1 h. SF reduced the friction coefficient by a factor of approximately 1.5 relative to saline. However, interstitial fluid pressurization was far more effective, reducing the friction coefficient by a factor of approximately 60 relative to equilibrium (zero-pressure) conditions. It was confirmed that SF acts as a boundary lubricant. CONCLUSIONS: These results emphasize the importance of interstitial fluid pressurization on the frictional response of cartilage. They imply that the mechanical integrity of cartilage must be maintained to produce low friction in articular joints. The more limited effectiveness of SF implies that intra-articular injections of lubricants in degenerated joints may have only limited effectiveness on their tribological properties. PMID- 18395477 TI - Intensive lipid-lowering therapy ameliorates novel calcification markers and GSM score in patients with carotid stenosis. AB - OBJECTIVES/DESIGN: Carotid plaque echogenicity quantified by the Gray-Scale Median (GSM) score has been associated with plaque vulnerability. The aim of this study was to assess whether intensive lipid-lowering treatment with atorvastatin in patients with carotid artery stenosis ameliorates novel vascular calcification inhibitors, such as osteopontin (OPN) and osteoprotegerin (OPG), and improves GSM score. METHODS: Ninety-seven patients with carotid stenosis (>40%), but without indication for intervention, were treated for 6 months with atorvastatin (10mg 80mg) to target LDL<100mg/dl. Fifty-two age-and sex-matched healthy individuals served as the control group. Blood samples and GSM were obtained at the beginning and after 6 months. RESULTS: Systolic blood pressure, hsCRP, fibrinogen, OPN and OPG levels differed significantly between patients with carotid stenosis and healthy controls at baseline (p<0.05). Atorvastatin treatment improved lipid profile and significantly reduced hsCRP (p=0.002), WBC count (p=0.041), OPN (p<0.001) and OPG levels (p<0.001). GSM score increased considerably after atorvastatin therapy (from 58.33+/-24.38 to 79.33+/-22.3; p<0.001) and that effect appeared related to OPN (p=0.001), OPG (p=0.013) and LDL (p=0.01) reduction. CONCLUSIONS: In patients with carotid stenosis, intensive lipid lowering therapy with statins attenuates serum OPN and OPG levels and enhances carotid plaque echogenicity, outlining their beneficial effects on plaque stability. PMID- 18395478 TI - Implications of a poroelastic cytoplasm for the dynamics of animal cell shape. AB - Two views have dominated recent discussions of the physical basis of cell shape change during migration and division of animal cells: the cytoplasm can be modeled as a viscoelastic continuum, and the forces that change its shape are generated only by actin polymerization and actomyosin contractility in the cell cortex. Here, we question both views: we suggest that the cytoplasm is better described as poroelastic, and that hydrodynamic forces may be generally important for its shape dynamics. In the poroelastic view, the cytoplasm consists of a porous, elastic solid (cytoskeleton, organelles, ribosomes) penetrated by an interstitial fluid (cytosol) that moves through the pores in response to pressure gradients. If the pore size is small (30-60nm), as has been observed in some cells, pressure does not globally equilibrate on time and length scales relevant to cell motility. Pressure differences across the plasma membrane drive blebbing, and potentially other type of protrusive motility. In the poroelastic view, these pressures can be higher in one part of a cell than another, and can thus cause local shape change. Local pressure transients could be generated by actomyosin contractility, or by local activation of osmogenic ion transporters in the plasma membrane. We propose that local activation of Na(+)/H(+) antiporters (NHE1) at the front of migrating cells promotes local swelling there to help drive protrusive motility, acting in combination with actin polymerization. Local shrinking at the equator of dividing cells may similarly help drive invagination during cytokinesis, acting in combination with actomyosin contractility. Testing these hypotheses is not easy, as water is a difficult analyte to track, and will require a joint effort of the cytoskeleton and ion physiology communities. PMID- 18395479 TI - Acupuncture and dry needling in the management of myofascial trigger point pain: a systematic review and meta-analysis of randomised controlled trials. AB - Pain from myofascial trigger points is often treated by needling, with or without injection, although evidence is inconclusive on whether this is effective. We aimed to review the current evidence on needling without injection, by conducting a systematic literature review. We searched electronic databases to identify relevant randomised controlled trials, and included studies where at least one group were treated by needling directly into the myofascial trigger points, and where the control was either no treatment, or usual care; indirect local dry needling or some form of placebo intervention. We extracted data on pain, using VAS scores as the standard. Seven studies were included. One study concluded that direct dry needling was superior to no intervention. Two studies, comparing direct dry needling to needling elsewhere in the muscle, produced contradictory results. Four studies used a placebo control and were included in a meta analysis. Combining these studies (n=134), needling was not found to be significantly superior to placebo (standardised mean difference, 14.9 [95%CI, 5.81 to 33.99]), however marked statistical heterogeneity was present (I(2)=88%). In conclusion, there is limited evidence deriving from one study that deep needling directly into myofascial trigger points has an overall treatment effect when compared with standardised care. Whilst the result of the meta-analysis of needling compared with placebo controls does not attain statistically significant, the overall direction could be compatible with a treatment effect of dry needling on myofascial trigger point pain. However, the limited sample size and poor quality of these studies highlights and supports the need for large scale, good quality placebo controlled trials in this area. PMID- 18395480 TI - Efficacy and safety of individualized growth hormone treatment in adult Japanese patients with growth hormone deficiency. AB - OBJECTIVE: The aim of the study was to evaluate the efficacy and safety of growth hormone (GH) treatment in Japanese adult patients with GH-deficiency. In the extension of the efficacy study, the effect of individualized-dosing (ID), based on insulin-like growth factor-I (IGF-I) levels, and fixed-dose (FD) GH regimens on body composition, were compared in Japanese GH-deficient adults. DESIGN: Randomized, double-blind (DB), placebo-controlled, 24-week treatment period followed by 48-week, open-label study in 43 endocrinology clinics in Japan. Patients received DB treatment with GH (0.012 mg/kg/day; n=57) or placebo (n=60) followed by open-label GH in an ID (n=75) or FD (0.012 mg/kg/day; n=38) regimen. SUBJECTS: Adult Japanese GH-deficient patients (peak GH<3 ng/mL). MEASUREMENTS: Trunk and total body fat (BF), lean body mass (LBM), and adverse events were determined. RESULTS: Percentage trunk fat was reduced significantly more in GH- than in placebo-treated patients at 24 weeks (-16.2 vs. 1.7%, p<0.0001). Open label treatment with an ID or FD GH regimen provided similar reductions in percentage trunk fat (-8.12 vs. -9.35%), and total BF (-0.92 vs. -0.70 kg) and a comparable increase in LBM (1.032 vs. 0.97 kg). Mean+/-SD GH doses (mg/kg/day) at 48 weeks were significantly lower with the ID GH regimen (ID, 0.0082+/-0.0050; FD, 0.0095+/-0.0033; p<0.05). The safety profile was comparable between ID and FD groups. CONCLUSIONS: Treatment with GH was associated with a significant reduction in trunk fat and improvement in serum lipid profile in Japanese adult GH-deficient patients. The improvement in body composition and tolerability were comparable between ID and FD GH regimens despite a significantly lower daily GH dose with the ID regimen. PMID- 18395481 TI - Clinical, ethical and legal considerations in the treatment of newborns with non ketotic hyperglycinaemia. AB - Non-ketotic hyperglycinaemia (NKH) is a devastating neurometabolic disorder leading, in its classical form, to early death or severe disability and poor quality of life in survivors. Affected neonates may need ventilatory support during a short period of respiratory depression. The transient dependence on ventilation dictates urgency in decision-making regarding withdrawal of therapy. The occurrence of patients with apparent transient forms of the disease, albeit rare, adds uncertainty to the prediction of clinical outcome and dictates that the current practice of withholding or withdrawing therapy in these neonates be reviewed. Both bioethics and law take the view that treatment decisions should be based on the best interests of the patient. The medical-ethics approach is based on the principles of non-maleficence, beneficence, autonomy and justice. The law relating to withholding or withdrawing life-sustaining treatment is complex and varies between jurisdictions. Physicians treating newborns with NKH need to provide families with accurate and complete information regarding the disease and the relative probability of possible outcomes of the neonatal presentation and to explore the extent to which family members are willing to take part in the decision making process. Cultural and religious attitudes, which may potentially clash with bioethical and juridical principles, need to be considered. PMID- 18395483 TI - A febrile traveler: mixed infection with waterborne organisms. PMID- 18395482 TI - Treatment outcomes and relapses of pulmonary tuberculosis in Lazio, Italy, 1999 2001: a six-year follow-up study. AB - OBJECTIVES: The aim of this study was to enhance tuberculosis (TB) treatment outcome monitoring by linking diverse surveillance systems and estimating treatment outcomes including relapse. METHODS: Tuberculosis treatment was surveyed in the Lazio region (Italy) from 1999 to 2001; a six-year follow-up of notified cases was undertaken to detect relapses. The results were analyzed as a population-based case-control study comparing each unsuccessful outcome and relapse with eligible controls. RESULTS: Of the 974 patients who entered the survey, 805 (82.6%) had complete treatment evaluations; 398 (49.4%) had a successful outcome, 401 (49.8%) had an unsuccessful outcome, and six developed chronic TB. Death was associated with age >64 years (OR 5.9; 95% CI 3.1-11.2), male gender (OR 2.1; 95% CI 1.0-4.4), and using second-line drugs (OR 2.3; 95% CI 1.0-5.4). Treatment failure was associated with previous treatment (OR 3.0; 95% CI 1.4-6.7) and being male, being foreign born (OR 6.6; 95% CI 2.1-21.2), receiving second-line drugs (OR 7.4; 95% CI 1.8-29.5), and receiving modified therapy (OR 5.1; 95% CI 1.7-14.9). Relapses after successful outcomes were detected in 5.5%, for which the strongest predictor was having extrapulmonary lesions (OR 22.8; 95% CI 1.8-287.3). CONCLUSIONS: Linking our survey data to other surveillance systems improved the mortality estimates and detected a high rate of relapse. Having received previous treatment and being a foreigner were independent determinants of treatment failure, suggesting that both acquired and primary drug resistance affect TB patients in Lazio. PMID- 18395484 TI - Failed DBS for palliation of visual problems in a case of oculopalatal tremor. AB - OBJECTIVE: To report the results of attempted bilateral red nucleus (RN) deep brain stimulation (DBS) for the palliative treatment of visual problems associated with oculopalatal tremor (OPT). BACKGROUND: It is hypothesized that OPT results from a defect in the Guillain-Mollaret triangle, a circuit that includes connections with the dentate nucleus, the contralateral red nucleus, and the inferior olive. We present a high functioning patient (an accountant) who underwent a palliative trial of RN region DBS in an approach targeted through the subthalamic nucleus region. The aim was to reduce eye tremor and improve vision through interruption of the pathologically oscillating circuit in the Guillain Mollaret triangle. METHODS: Following informed consent, a patient with OPT (and failure of multiple classes of medication and botulinum toxin therapy) underwent placement of bilateral DBS electrodes within the region of the RN. He underwent preoperative testing and testing after 12 months of continuous stimulation with the device in monopolar, bipolar, low frequency, and high frequency settings. RESULTS: The patient did not demonstrate significant changes in the neurological examination following the procedure and postoperative programming sessions. Eye tremor was monitored pre- and postoperatively by ocular EMG and did not change in frequency. Following the one-year trial, stimulation was discontinued as there were no improvements in vision. CONCLUSION: DBS for OPT was not clinically effective. There were many potential reasons for failed efficacy including a failure to implant the electrodes deep and medial enough into the target region because of stimulation induced side effects. Other targets within the Guillain Mollaret circuit (and outside of the circuit) may be more useful, though they may prove to be less safe and even more difficult to access. Better custom designed DBS leads may be needed for such small targets in critical brain regions. PMID- 18395485 TI - Proteases and antiproteases in immune defense, tissue homeostasis and development. PMID- 18395486 TI - Hyper-responsiveness of IPF/UIP fibroblasts: interplay between TGFbeta1, IL-13 and CCL2. AB - One of the hallmarks of idiopathic pulmonary fibrosis with a usual interstitial pneumonia histological pathology (IPF/UIP) is excess collagen deposition, due to enhanced fibroblast extracellular matrix synthetic activity. Studies using murine models of lung fibrosis have elucidated a pro-fibrotic pathway involving IL-13 driving CCL2, which in turn drives TGFbeta1 in lung fibroblasts. Therefore, we sought to determine whether this pathway exists in the human fibrotic setting by evaluating human IPF/UIP fibroblasts. IPF/UIP fibroblasts have an increased baseline fibrotic phenotype compared to non-fibrotic fibroblasts. Interestingly, non-fibrotic fibroblasts responded in a pro-fibrotic manner to TGFbeta1 but were relatively non-responsive to IL-13 or CCL2, whereas, IPF/UIP cells were hyper responsive to TGFbeta1, IL-13 and CCL2. Interestingly, TGFbeta1, CCL2 and IL-13 all upregulated TGFbeta receptor and IL-13 receptor expression, suggesting an ability of the mediators to modulate the function of each other. Furthermore, in vivo, neutralization of both JE and MCP5, the two functional orthologs of CCL2, during bleomycin-induced pulmonary fibrosis significantly reduced collagen deposition as well as JE and CCR2 expression. Also in the bleomycin model, CTGF, which is highly induced following TGFbeta stimulation, was attenuated with anti JE/anti-MCP5 treatment. Overall this study demonstrates an interplay between TGFbeta1, IL-13 and CCL2 in IPF/UIP, where these three mediators feedback on each other, promoting the fibrotic response. PMID- 18395487 TI - Properties, intracellular localization, and stage-specific expression of membrane bound beta-glucosidase, BglM1, from Physarum polycephalum. AB - Physarum polycephalum expresses a membrane-bound beta-glucosidase (BglM1) with a molecular mass of 130 kDa. The primary structure of BglM1 consists of a glycosyl hydrolase family 3 domain at an amino-terminal domain and a carboxyl-terminal region without homology to the sequence of known glycosidases. The latter region contains two calx-beta motifs known as Ca(2+)-binding sites; an RGD sequence, which is known to be a cell attachment sequence; and a transmembrane region. The molecular mass calculated from the amino acid sequence is 130 kDa, but that in the crude extract was estimated by SDS-PAGE to be 230 kDa, and decreased to 130 kDa during purification. However, when BglM1 was purified in the presence of calcium ion, the molecular mass remained 230 kDa. The biochemical characteristics of the 130- and 230-kDa BglM1 forms were analyzed: differences were found in the kinetic data for some substrates specific for both these enzymes; however, no difference was found in their intrinsic characteristics such as optimum pH and temperature. In addition, the molecular mass of native BglM1 with a calcium ion was estimated to be 1,000 kDa or larger by gel filtration. These results suggest that the calcium ion influences the conformation of BglM1. The evidence that BglM1 localizes on the plasma membrane of plasmodia was confirmed using immunofluorescence microscopy. Although Physarum BglM1 was expressed in microplasmodia and plasmodia, little expression was detected in other stages. BglM1 may have some function only in multinuclear cells. PMID- 18395489 TI - Novel receptor kinases involved in growth regulation. AB - The growth of plant cells involves a constant adjustment of synthesis and rearrangement of cell wall polymers. Recently, three plasma membrane-bound receptor kinases related to CrRLK1 have been shown to be involved in the negative control of cell growth in different contexts. THESEUS1 is activated in mutants deficient for cellulose and may act as a cell wall integrity sensor inhibiting cell elongation. FERONIA is polarly localized in synergid cells of the female gametophyte and is required for growth cessation of compatible pollen tubes and subsequent delivery of sperm cells. AmRLK is involved in the control of the polar conical outgrowth of epidermal cells of Antirrhinum petals. The conservation of both extracellular and kinase domains suggests that the three receptors bind to related ligands and have similar cellular outputs, which may involve the production of reactive oxygen species. PMID- 18395488 TI - Proteases and cystic fibrosis. AB - Cystic fibrosis is the most common, inherited fatal disease in Caucasians. The major cause of morbidity and mortality is chronic lung disease due to infection and inflammation in the airways leading to bronchiectasis and respiratory failure. The signature pathologic features of CF lung disease including abnormal mucus obstructing airways, chronic infection with Staphylococcus aureus, Pseudomonas aeruginosa and other gram negative bacteria, and a robust neutrophil dominant airway inflammation, are exacerbated by unopposed proteases present at high concentrations in the ASL. There is strong evidence that proteases, particularly neutrophil elastase, contribute to the pathology of CF by impairing mucociliary clearance, interfering with innate immune functions, and perpetuating neutrophilic inflammation. The mechanisms employed by proteases to impact airway function in CF will be reviewed. PMID- 18395490 TI - Biochemical properties of Cu/Zn-superoxide dismutase from fungal strain Aspergillus niger 26. AB - The fungal strain Aspergillus niger produces two superoxide dismutases, Cu/Zn-SOD and Mn-SOD. The primary structure of the Cu/Zn-SOD has been determined by Edman degradation of peptide fragments derived from proteolytic digests. A single chain of the protein, consisting of 153 amino acid residues, reveals a very high degree of structural homology with the amino acid sequences of other Aspergillus Cu/Zn SODs. The molecular mass of ANSOD, measured by MALDI-MS and ESI-MS, and calculated by its amino acid sequence, was determined to be 15821 Da. Only one Trp residue, at position 32, and one disulfide bridge were identified. However, neither a Tyr residue nor a carbohydrate chain occupying an N-linkage site (-Asn Ile-Thr-) were found. Studies on the temperature and pH dependence of fluorescence, and on the temperature dependence of CD spectroscopic properties, confirmed that the enzyme is very stable, which can be explained by the stabilising effect of the disulfide bridge. The enzyme retains about 53% of its activity after incubation for a period of 30 min at 60 degrees C, and 15% at 85 degrees C. PMID- 18395491 TI - Umbilical cord blood transplantation: basic biology and clinical challenges to immune reconstitution. AB - Allogeneic stem cell transplantation has continued to evolve as a common procedure for the treatment of hematological malignancies and bone marrow failure. Donor bone marrow and mobilized peripheral stem cells are routinely employed for the reconstitution of immune function in leukemia and lymphoma patients following radiation and/or chemotherapy. Unfortunately, only 30% of patients have an HLA-identical sibling donor and the identification of matched unrelated donors, particularly for minorities, can present an exceptional challenge. The transplantation of umbilical cord blood (UCB) represents the most recent strategy to expand the potential donor pool while maintaining an acceptable level of treatment-related complications. First utilized in children, UCB transplantation permits a higher degree of HLA disparity while demonstrating a reduction in the incidence and severity of graft-versus-host disease (GvHD) compared to previous transplantation modalities. Despite the apparent decrease in GvHD, relapse rates remain comparable to transplantation with bone marrow or mobilized peripheral blood suggesting a strong graft-versus-leukemia/lymphoma (GvL) effect. However, several issues complicate the use of UCB transplantation and its extension to the treatment of adults. Many infections that afflict transplant patients are particularly frequent and more severe in the context of UCB transplantation. UCB T-cells are naive and therefore display less proliferation and IFN-gamma production in response to cognate antigen and also appear to demonstrate defects in signal transduction mechanisms. In addition, UCB contains T regulatory cells (Treg) with more potent suppressor function than adult Treg. Furthermore, adult patients often require more total cells and CD34+ progenitors for transplantation than a single UCB unit can provide. Thus, strategies to expand selected subpopulations from UCB and the use of multi-unit transplantation are areas of active research. This review will provide a condensed summary of the clinical history of UCB transplantation and emphasize the advantages and disadvantages of this approach to hematological malignancies in comparison to other methods of hematopoietic stem cell transplantation. Subsequently, it will mainly focus on the current challenges to immune reconstitution presented by UCB transplantation, recent research into their cellular and molecular mechanisms, and experimental approaches to overcome them. PMID- 18395492 TI - The Src/ABL kinase inhibitor dasatinib (BMS-354825) inhibits function of normal human T-lymphocytes in vitro. AB - Dasatinib (BMS-354825) is a Src/ABL tyrosine kinase inhibitor currently approved for the treatment of chronic myeloid leukemia. Dasatinib has increased potency against ABL compared to the current therapy imatinib, and is effective in many cases where disease is resistant to imatinib. Dasatinib also inhibits many Src family tyrosine kinases. We have demonstrated in this study that dasatinib is able to block the function of normal human T-lymphocytes in vitro at clinically relevant concentrations. T-cell functions including proliferation, activation and cytokine production were all uniformly inhibited in the presence of dasatinib. We also demonstrated inhibition of TCR signalling through Src-family kinase LCK, and predicted that inhibition of LCK and other kinases involved in T-cell signalling by dasatinib is responsible for the suppression of T-cell function. These findings raise the concern about potential T-cell inhibition in patients taking dasatinib, and suggest a possible application for the treatment of T-cell mediated immune disorders. PMID- 18395493 TI - Conventional myomectomy. AB - In addition to the conventional/older treatments of myomectomy and hysterectomy, the options now available to the woman with symptomatic fibroids, especially if she wishes to conserve her uterus, include medical treatments such as mifepristone, minimally invasive therapies such as uterine artery embolization (UAE) or magnetic-resonance-guided focused ultrasound surgery (MRgFUS), and laparoscopic or vaginal myomectomy. It is generally accepted, and with justification, that conventional myomectomy is associated with significant morbidity, especially excessive peri-operative blood loss, recurrence of the fibroids and adhesion formation, which might compromise the very reason, i.e. fertility, which the operation is performed to preserve. However, the newer treatments have significant limitations: medical treatments are promising but, to date, have been found to be of limited efficacy; UAE is still under evaluation and its impact on fertility has yet to be researched; and MRgFUS is an even newer therapy which is limited to centres with high technology and hugely expensive open magnetic resonance imaging facilities. Both UAE and MRgFUS cause shrinkage rather than removal of the fibroids, and have limited efficacy when used with really large, multiple fibroids. Laparoscopic myomectomy is also limited by the size and number of fibroids that can be treated by this approach, and demands laparoscopic skills that are still lacking in most institutions; limitations which also apply to vaginal myomectomy. It is therefore evident that conventional abdominal myomectomy still has a major role to play. There are no limitations on size and number of fibroids, and there are good data showing improvement in outcomes of assisted reproduction treatments following myomectomy. The widespread fallacy is probably the assumption that any gynaecological surgeon can perform a myomectomy; good conventional myomectomy demands no less skill than the laparoscopic approach. There is a need to continue to refine and innovate, especially with regard to reducing blood loss during surgery, reducing the risk of adhesion formation, reducing the risk of recurrence, and reconstruction of uteri to approximate anatomical normality and physiological integrity so that they can carry a pregnancy without complications such as scar rupture. This chapter will review the position of conventional myomectomy and describe approaches to optimizing outcomes following myomectomy. PMID- 18395494 TI - Segmentation of cDNA microarray images by kernel density estimation. AB - The segmentation of cDNA microarray spots is essential in analyzing the intensities of microarray images for biological and medical investigation. In this work, nonparametric methods using kernel density estimation are applied to segment two-channel cDNA microarray images. This approach groups pixels into both a foreground and a background. The segmentation performance of this model is tested and evaluated with reference to 16 microarray data. In particular, spike genes with various contents are spotted in a microarray to examine and evaluate the accuracy of the segmentation results. Duplicated design is implemented to evaluate the accuracy of the model. The results of this study demonstrate that this method can cluster pixels and estimate statistics regarding spots with high accuracy. PMID- 18395495 TI - An ontology-driven semantic mashup of gene and biological pathway information: application to the domain of nicotine dependence. AB - OBJECTIVES: This paper illustrates how Semantic Web technologies (especially RDF, OWL, and SPARQL) can support information integration and make it easy to create semantic mashups (semantically integrated resources). In the context of understanding the genetic basis of nicotine dependence, we integrate gene and pathway information and show how three complex biological queries can be answered by the integrated knowledge base. METHODS: We use an ontology-driven approach to integrate two gene resources (Entrez Gene and HomoloGene) and three pathway resources (KEGG, Reactome and BioCyc), for five organisms, including humans. We created the Entrez Knowledge Model (EKoM), an information model in OWL for the gene resources, and integrated it with the extant BioPAX ontology designed for pathway resources. The integrated schema is populated with data from the pathway resources, publicly available in BioPAX-compatible format, and gene resources for which a population procedure was created. The SPARQL query language is used to formulate queries over the integrated knowledge base to answer the three biological queries. RESULTS: Simple SPARQL queries could easily identify hub genes, i.e., those genes whose gene products participate in many pathways or interact with many other gene products. The identification of the genes expressed in the brain turned out to be more difficult, due to the lack of a common identification scheme for proteins. CONCLUSION: Semantic Web technologies provide a valid framework for information integration in the life sciences. Ontology driven integration represents a flexible, sustainable and extensible solution to the integration of large volumes of information. Additional resources, which enable the creation of mappings between information sources, are required to compensate for heterogeneity across namespaces. RESOURCE PAGE: http://knoesis.wright.edu/research/lifesci/integration/structured_data/JBI-2008/ PMID- 18395496 TI - Role of respiratory system impedance in the difference of ventilatory control between children and adults. AB - How children are able to adapt their ventilation to the intensity of exercise faster than adults remain unclear. We hypothesized that differences of VE observed between children and adults depend on either peripheral chemoreceptors or central command activity. We examined ventilatory control parameters in either normoxic or hypoxic condition (FI 02 =0.15). We analyzed the adaptability of the respiratory exchanges by (i) the measurement of ventilatory kinetics time constant and (ii) the central command by the mouth-occlusion pressure (P0.1). A group of nine pre-pubescent children (mean age 9.5+/-1 years) and a group of eight adults (mean age 24+/-3.1 years) performed a constant-load exercise. In normoxia, children had significantly shorter time-constant (tau) VCO2 (respectively, 38.5+/-4.3 and 53.1+/-5.3s; P<0.001), tau VE (respectively, 52.5+/ 13.1s vs. 66.1+/-12.3s; P<0.001), and tau P0.1 (57.4+/-15.4 and 61.0+/-12.9s, respectively; P<0.001) than adults. In hypoxia, children exhibited shorter tau P0.1/VT/Ti compare to adults. Reinforced by the significant correlation between tau VE and tau P0.1/VT/Ti for children but not adults, we concluded that ventilatory response differences could be due in part to the respiratory system impedance. PMID- 18395497 TI - Glutamatergic and purinergic mechanisms on respiratory modulation in the caudal NTS of awake rats. AB - In the present study we evaluated the role of ionotropic glutamate receptors and purinergic P2 receptors in the caudal commissural NTS (cNTS) on the modulation of the baseline respiratory frequency (fR), and on the tachypneic response to chemoreflex activation in awake rats. The selective antagonism of ionotropic glutamate receptors with kynurenic acid (2 nmol/50 nl) in the cNTS produced a significant increase in the baseline fR but no changes in the tachypneic response to chemoreflex activation. The selective antagonism of purinergic P2 receptors by PPADS (0.25 nmol/50 nl) in the cNTS produced no changes in the baseline fR or in the tachypneic response to chemoreflex activation. The data indicate that glutamate acting on ionotropic receptors in the cNTS plays a inhibitory role on the modulation of the baseline fR but had no effect on the tachypneic response to chemoreflex activation, while ATP acting on P2 receptors in the cNTS plays no major role in the modulation of the baseline fR or in the tachypneic response to chemoreflex activation. We suggest that neurotransmitters other than l-glutamate and ATP are involved in the processing of the tachypneic response of the chemoreflex at the cNTS level. PMID- 18395498 TI - Cytoskeletal mechanics in airway smooth muscle cells. AB - Mechanical properties and contractility of airway smooth muscle tissue are largely responsible for airway narrowing and airway hyperresponsiveness in asthma. To explain these pathological phenomena, investigators have studied the mechanical behaviour of airway smooth muscle cells and its relationship to the underlying cellular biophysical and biochemical mechanisms. During the past decade, a growing body of evidence has indicated that a deformable intracellular polymer network, known as the cytoskeleton, plays a major role in transmitting and distributing mechanical forces within the cell and in their conversion into biochemical responses. We review here evidence suggesting that the tensed and crosslinked cytoskeletal lattice, the contractile apparatus, and the cytoskeleton extracellular matrix interactions are key determinants of mechanical properties and mechanosensing of airway smooth muscle cells, with the mechanical distending stress of the cytoskeleton playing the central role. PMID- 18395499 TI - Simultaneous measurement of tryptophan and related compounds by liquid chromatography/electrospray ionization tandem mass spectrometry. AB - We have expanded a liquid chromatographic-tandem mass spectrometric method that measures 3-hydroxykynurenine and 3-hydroxyanthranilic acid in addition to tryptophan and kynurenine both intra- and extracellularly. After reversed phase HPLC separation, the compounds were detected in the MS positive multiple reaction monitoring mode. We found a good linear response for each tryptophan metabolite. The lower limit of quantification for each compound ranged from 0.01 to 0.1 microM. The extraction efficiencies from spiked cell samples and culture medium ranged between 83 and 111% and the overall coefficient of variation of analyses was less than 7%. Using our method, we found tryptophan metabolites in the cells and the culture medium of LN229 human glioma cells were stimulated by interferon gamma, a known inducer of indoleamine 2,3-dioxygenase. The intracellular concentrations of kynurenine, 3-hydroxykynurenine and 3-hydroxyanthranilic acid were higher than those in the medium. This is the first report of a method for the simultaneous determination of tryptophan and its metabolic products both intra- and extracellularly. PMID- 18395501 TI - Consensus recommendations for managing asymptomatic persistent non-virus non alcohol related elevation of aminotransferase levels: suggestions for diagnostic procedures and monitoring. AB - A persistent increase in non-virus non-alcohol related aminotransferase levels can have multiple causes, which differ in terms of prevalence and clinical importance. In the general population, the most frequent cause is non-alcoholic hepatic steatosis, which can evolve into steato-hepatitis and cirrhosis. The treatment for steatosis and non-alcoholic steato-hepatitis consists of modifying lifestyles, whereas the effectiveness of drug treatment remains to be determined. Other much less frequent (yet not rare) causes of persistent non-virus non alcohol related elevations in aminotransferase levels are celiac disease and hemochromatosis, whereas autoimmune hepatitis, primary biliary cirrhosis, primary sclerosing cholangitis, and alpha-1-anti-trypsin deficit are rare. Given that some of these conditions are susceptible to treatment, early diagnosis is important. No epidemiological data are available for evaluating the prevalence of elevated aminotransferase levels correlated with the toxicity of drugs or other xenobiotics, including herbal products. The present document, created by a panel of experts based on a systematic review of scientific evidence, is mainly geared towards physicians working in General Medicine and Transfusion Centres, who generally represent the first contact of persons with elevated aminotransferase levels. The document includes suggestions for diagnosing causes of persistent non virus non-alcohol related increases in aminotransferase levels, considering the frequency and response to treatment. The conditions requiring specialized visits are also indicated. PMID- 18395503 TI - The relationship between breast cancer and breast augmentation with injected polyacrylamide gel: two case reports. PMID- 18395502 TI - Estrogens stimulate the proliferation of human cholangiocarcinoma by inducing the expression and secretion of vascular endothelial growth factor. AB - BACKGROUND: Estrogens may induce the proliferation of neoplastic cells by activating neo-angiogenesis. AIM: To evaluate the effect of estrogens on the expression of vascular endothelial growth factor (VEGF) and related receptors (VEGF-R) in human cholangiocarcinoma and the role played by VEGF in mediating the proliferative effects of estrogens. METHODS: Seven biopsies of intra-hepatic cholangiocarcinoma and the HuH-28 cell lines were investigated. Cell proliferation was measured by both PCNA Western blot and MTS proliferation assay. RESULTS: By immunohistochemistry, biopsies of human cholangiocarcinoma stained positively for VEGF-A and VEGF-C and related receptors. HuH-28 cells expressed VEGF-A, -C, and VEGFR-1, -2, -3 and, their protein level was enhanced by 17beta estradiol in association with the stimulation of cell proliferation. 17beta Estradiol-stimulated proliferation of HuH-28 cells was blocked by 70% by VEGF TRAP, a receptor-based VEGF inhibitor. 17beta-Estradiol induced the secretion of VEGF in the supernatant of HuH-28 cells. The stimulatory effect of 17beta estradiol on the protein expression of VEGF-A, VEGF-C and VEGFR-1, -2, -3 was blocked by antagonists of ER (Ici182,780) or insulin-like growth factor 1 receptor (alphaIR3). CONCLUSIONS: With the limitations of experiments performed in a cell line, our study indicates that VEGF plays a major role in mediating the proliferative effects of estrogens on human cholangiocarcinoma. PMID- 18395504 TI - Computer-based screening of chest X-rays for vertebral compression fractures as an osteoporosis index in men. AB - We evaluated the recognition of osteoporosis in the veteran male population through a computer-based review of chest X-ray (CXR) reports in the Veterans Affairs Northern California Health Care System database, looking for unrecognized vertebral fractures. All CXR reports between January 1, 2000 and December 31, 2001, were scanned for the terms "compression" or "wedg (where the "" indicates a wild card search encompassing such terms as "wedge" or "wedging")". During this time, 26,994 CXR examinations were performed on 18,069 patients. 22,494 (83.3% of the total) CXR examinations were done in 14,561 men >or=50 years of age. 780 CXR reports (3.5%) encompassing 664 men (4.5%) contained at least one key phrase suggesting osteoporosis. Three years later, 495 of these 664 men were still living. 99 of these (20%) had been diagnosed with osteoporosis, 72 (15%) had a dual-energy X-ray absorptiometry (DXA) scan, and 89 (18%) had ever been prescribed a bisphosphonate. Overall, only 126 (25%) men had chart documentation indicating some recognition by the provider of the abnormality reported on CXR. We conclude that a significant fraction of men >50 years old may have unrecognized osteoporosis severe enough to result in vertebral fracture. We conclude that computerized screening of CXR reports may represent an effective strategy to aid clinicians in identifying men at risk for further debilitating fractures. PMID- 18395505 TI - On-gel fluorescent visualization and the site identification of S-nitrosylated proteins. AB - Mounting evidence indicates that S-nitrosylation of critical cysteine residues in a protein represents a common feature of protein function regulation and cell signaling. However, the progress in studying the exact role of S-nitrosylation has been hampered by a lack of rapid and accurate methods for the detection of these S-nitrosylated proteins and the exact modification sites. In this article, we report a fluorescence-based method in which the S-nitrosylated cysteines are converted into 7-amino-4-methylcoumarin-3-acetic acid (AMCA) fluorophore-labeled cysteines-termed the AMCA switch method. The labeled proteins are then analyzed by nonreducing SDS-PAGE, and the S-nitrosylated proteins can be readily detected as brilliant blue bands after the activation of ultraviolet light. Furthermore, the sites of modification can be determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) after in-gel tryptic digestion of the fluorescent band, and the recognizable AMCA tag in the MS spectra ensures the accurate site identification of the nitrosocysteines. Therefore, our method offers some apparent advantages by direct visualization of on-gel image of S-nitrosylated proteins, shorter experiment time by skipping the anti-biotin immunoblotting step, and elimination of the potential interference of endogenous biotinylated proteins. Based on this method, we detected the S-nitrosylation and the modified site in bovine serum albumin and gankyrin after in vitro S-nitrosylation. Overall, our results indicate that the AMCA switch method is a fast and accurate method to identify the S-nitrosylated protein and the modification sites. PMID- 18395500 TI - Colonoscopy practice in Italy: a prospective survey on behalf of the Italian Association of Hospital Gastroenterologists. AB - BACKGROUND: Examining the current practice is important for the benchmarking of quality of colonoscopy and the comparison with the standards and recommendations expected by professional societies. AIM: To describe colonoscopy practice in Italy, on the basis of prospective analysis of a large number of examinations performed by operators with different levels of expertise in a wide range of unselected centres. DESIGN AND SETTING: Cross-sectional, prospective and multicentre study. METHODS: The main features of each endoscopy centre (structure indicators) were collected through the use of a standardised questionnaire. A second questionnaire was used to prospectively record details of all the consecutive colonoscopies performed in a 2-week study period. RESULTS: Data from 278 centres and 12,835 consecutive colonoscopies were evaluated. Centres were uniformly distributed throughout Italy - north, centre and south - as was their organizational complexity and workload. Overall, adequate facilities (i.e. cleaning area for disinfection/reprocessing, equipped recovery room), and safety equipment (i.e. pulse oximetry, equipment for emergency cardiopulmonary resuscitation) were lacking in a considerable amount of centres, especially in those with a lower degree of organizational complexity. Written informed consent was routinely required in 87% of the centres, but a specific consent for colonoscopy, including detailed information on adverse events, was adopted by 66%. Regular programs for recording some quality indicators (i.e. cecal intubation, quality of bowel cleansing, patients' satisfaction and complications) were implemented in a minority of centres. About 93% of the colonoscopies were performed for diagnostic purpose; screening and surveillance accounted for 13.7% and 25.3% of the indications, respectively. Sedation and/or analgesia was administered in about half of the patients. Overall, colonoscopies were completed to the cecum in 80.7% of cases, and only 22.1% of the centres reported a cecal intubation rate >or=90%; this figure was reported in 33.1% of centres with high organizational complexity and in 14.4% of outpatient units. The overall incidence of immediate complications was low (2 perforations, 26 bleedings and 32 serious cardiorespiratory complications). CONCLUSIONS: This study documented a wide variation in colonoscopy practice between centres and highlighted problem areas where interventions are needed to improve performance and safety of the examinations. PMID- 18395506 TI - Spectral resolution of a second binding site for Nile Red on cytochrome P4503A4. AB - Nile Red is sequentially metabolized by cytochrome P4503A4 to the N-monoethyl and N-desethyl products, which typifies the metabolism of many amine containing drugs. Sequential metabolism of a single substrate results in complex kinetics that confound predictive models of drug clearance. As a fluorescent model for drugs which undergo sequential metabolism, Nile Red provides the opportunity to monitor drug-CYP interactions wherein the fluorescent properties of Nile Red could, in principle, be exploited to determine individual rate and equilibrium constants for the individual reactions. Previously, it was shown that Nile Red binds at the active site and fluoresces (K(D) approximately 50nM) with maximum emission at approximately 620nm, but it was unclear whether a red-shifted emission, at approximately 660nm, consisted of only free Nile Red or Nile Red bound at a second site on the protein. Here, equilibrium binding studies, including 'reverse titrations' spanning low ratios of CYP3A4/Nile Red, indicate two binding sites for Nile Red with a contribution to the 'red emission' greater than can be accounted for by free Nile Red. Singular value decomposition affords basis spectra for both spectral components and fits well to the experimentally determined concentration dependence of Nile Red emission. In addition, the red spectral component, with an apparent K(D)=2.2muM, is selectively eliminated by titration with the known allosteric effectors of CYP3A4, alpha-napthoflavone and testosterone. Furthermore, the double mutant L2311F/D214E, previously demonstrated to perturb a peripheral allosteric site, lacks the red-emitting Nile Red binding site, but retains the blue-emitting site. Together these data indicate that a second Nile Red site competes with other effectors of CYP3A4 at a site that results in Nile Red emission at 660nm. PMID- 18395507 TI - Sphingolipids and cellular cholesterol homeostasis. Effect of ceramide on cholesterol trafficking and HMG CoA reductase activity. AB - We previously showed that degradation of cellular sphingomyelin (SM) by SMase C results in a greater stimulation of cholesterol translocation to endoplasmic reticulum, compared to its degradation by SMase D. Here we investigated the hypothesis that the effect of SMase C is partly due to the generation of ceramide, rather than due to depletion of SM alone. Inhibition of hydroxymethylglutaryl CoA reductase (HMGCR) activity was used as a measure of cholesterol translocation. Treatment of fibroblasts with SMase C resulted in a 90% inhibition of HMGCR, whereas SMase D treatment inhibited it by 29%. Treatment with exogenous ceramides, or increasing the endogenous ceramide levels also inhibited HMGCR by 60-80%. Phosphorylation of HMGCR was stimulated by SMase C or exogenous ceramide. The effects of ceramide and SMase D were additive, indicating the independent effects of SM depletion and ceramide generation. These results show that ceramide regulates sterol trafficking independent of cellular SM levels. PMID- 18395509 TI - Thyroid hormone receptor mediates human MDR1 gene expression-Identification of the response region essential for gene expression. AB - P-glycoprotein, encoded by the MDR1 gene, is a drug efflux transporter that is expressed in various tissues and plays an important role in the absorption and elimination of many drugs and xenobiotics. Induction of the MDR1 gene affects drug disposition and the efficacy of drug treatment. In this study, we demonstrated that the thyroid hormone receptor (TR) induces MDR1 gene expression in a thyroid hormone (TH)-dependent manner. The 5'-upstream region of the human MDR1 gene was examined for the presence of TH-responsive elements. Luciferase reporter gene assays revealed that the TH response region is located between -7.9 and -7.8kb upstream from the transcription start site of MDR1. The region contains two TH response clusters, one of which includes a direct repeat with a three-nucleotide spacer (DR3) and a four-nucleotide spacer DR4(I), and the other of which includes two DR4s (II and III). Mutation analyses indicated that every direct repeat has a unique contribution to the TH response. In particular, DR4(I) was shown to be the most important element. Chromatin immunoprecipitation assays revealed that TR and retinoid X receptor (RXR) bind to the TH response region, and gel mobility shift assays confirmed that one molecule of TR/RXR heterodimer binds to each of the clusters in this region, with preferential binding to the upstream one. We furthermore demonstrated that two molecules of TR/RXR could bind simultaneously to the TH response region. The order of binding affinity to the direct repeats was DR4(I)>DR4(II)>DR4(III) approximately DR3. Our results indicate that these two closely spaced TR/RXR-binding clusters are both required for the maximal induction of MDR1 gene expression mediated by TR. PMID- 18395510 TI - Chaperonins: The hunt for the Group II mechanism. AB - Chaperonins are multi-subunit complexes that enhance the efficiency of protein folding reactions by capturing protein substrates in their central cavities. They occur in all prokaryotic and eukaryotic cell types and, alone amongst molecular chaperones, chaperonin knockouts are always lethal. Chaperonins come in two forms; the Group I are found in bacteria, mitochondria and plastids [W.A. Fenton, A.L. Horwich, Q. Rev. Biophys. 36 (2003) 229-256, [1]] and the Group II in the eukaryotic cytoplasm and in archaea [N.J. Cowan, S.A. Lewis, Adv. Protein Chem. 59 (2001) 73-104, [2]]. Both use energy derived from ATP binding and hydrolysis to drive a series of structural rearrangements that enable them to capture, engulf and then release polypeptide chains that have either not yet acquired the native, biologically active state or have been denatured in the cell. PMID- 18395508 TI - Functions of RANKL/RANK/OPG in bone modeling and remodeling. AB - The discovery of the RANKL/RANK/OPG system in the mid 1990s for the regulation of bone resorption has led to major advances in our understanding of how bone modeling and remodeling are regulated. It had been known for many years before this discovery that osteoblastic stromal cells regulated osteoclast formation, but it had not been anticipated that they would do this through expression of members of the TNF superfamily: receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG), or that these cytokines and signaling through receptor activator of NF-kappaB (RANK) would have extensive functions beyond regulation of bone remodeling. RANKL/RANK signaling regulates osteoclast formation, activation and survival in normal bone modeling and remodeling and in a variety of pathologic conditions characterized by increased bone turnover. OPG protects bone from excessive resorption by binding to RANKL and preventing it from binding to RANK. Thus, the relative concentration of RANKL and OPG in bone is a major determinant of bone mass and strength. Here, we review our current understanding of the role of the RANKL/RANK/OPG system in bone modeling and remodeling. PMID- 18395511 TI - EGFR signalling is required for Paracentrotus lividus endomesoderm specification. AB - The EGFR pathway is critical for cell fate specification throughout the development of several organisms. Here we identified in sea urchin an EGFR related antigen maternally expressed and showing a dynamic pattern of localization during development. To investigate the role played by the EGFR in Paracentrotus lividus development we blocked its activity by using the EGFR kinase inhibitor AG1478. This treatment produces decrease of EGFR phosphorylation, and embryos with various defects especially in the endomesoderm territory until to obtain an animalized phenotype. These effects are rescued by the addition of TGF-alpha, an EGFR ligand. The role played by EGFR-like along the animal/vegetal axis was also detected, after AG1478 treatment, by the extended distribution of HE and decreased nuclearization of beta-catenin in vegetal cells. Moreover, inhibition of EGFR-like reduced ERK phosphorylation, necessary for cell fate specification in the micromeres and their derivates. Taken together these results indicate that EGFR-like activity is required both for A/V axis formation and endomesoderm differentiation. PMID- 18395512 TI - Diphenyleneiodonium acutely inhibits reactive oxygen species production by mitochondrial complex I during reverse, but not forward electron transport. AB - We investigated the effects of diphenyleneiodonium (DPI) on superoxide production by complex I in mitochondria isolated from rat skeletal muscle. Superoxide production was measured indirectly as hydrogen peroxide production. In a conventional medium containing chloride, DPI strongly inhibited superoxide production by complex I driven by reverse electron transport from succinate. In principle, this inhibition could be explained by an observed decrease in the mitochondrial pH gradient caused by the known chloride-hydroxide antiport activity of DPI. In a medium containing gluconate instead of chloride, DPI did not affect the pH gradient. In this gluconate medium, DPI still inhibited superoxide production driven by reverse electron transport, showing that the inhibition of superoxide production was not dependent on changes in the pH gradient. It had no effect on superoxide production during forward electron transport from NAD-linked substrates in the presence of rotenone (to maximise superoxide production from the flavin of complex I) or antimycin (to maximise superoxide production from complex III), suggesting that the effects of DPI were not through inhibition of the flavin. We conclude that DPI has the novel and potentially very useful ability to prevent superoxide production from the site in complex I that is active during reverse electron transport, without affecting superoxide production during forward electron transport. PMID- 18395513 TI - Inhibition of Bufo arenarum oocyte maturation induced by cholesterol depletion by methyl-beta-cyclodextrin. Role of low-density caveolae-like membranes. AB - The invaginated structure of caveolae seems to provide an optimal environment for hormone binding leading to oocyte meiotic maturation. We conducted a quantitative analysis of lipids and proteins of detergent-free low-density membranes isolated from Bufo arenarum oocytes and we modulated cellular cholesterol to further understand how these domains perform their regulatory functions in the amphibian system. Light membranes derive from the plasma membrane as suggested by the enrichment in the activity of 5'nucleotidase. Lipid analysis by chromatography techniques revealed that this fraction is enriched in phosphatidylserine and cholesterol and that it evidences an important level of sphingomyelin. The finding of a single 21 kDa caveolin in light membranes indicates the presence of caveolae-like structures in B. arenarum oocytes. In support of this finding, c Src is significantly associated to this fraction. Cholesterol content of oocytes treated with methyl-beta-cyclodextrin (MbetaCD) decreased when compared to control oocytes. Drug treatment inhibited meiotic maturation in a dose-dependent manner and affected the localization of caveolin and c-Src among membrane fractions. Repletion of cholesterol showed a recovery of the ability of MbetaCD treated oocytes to mature, particularly at the 25 mM concentration in which reversibility was close to the control level. Results highlight the importance of caveolae-like microdomains for maturation signaling in Bufo oocytes. PMID- 18395514 TI - Importance of the phosphocholine linkage on sphingomyelin molecular properties and interactions with cholesterol; a study with phosphate oxygen modified sphingomyelin-analogues. AB - We have characterized the molecular properties and membrane behavior of synthetically modified sphingomyelin analogues, modified on the oxygen connecting the phosphocholine group to the ceramide backbone. The oxygen was replaced with an S-atom (S-PSM), an NH-group (NH-PSM) or a CH(2)-group (CH(2)-PSM). Diphenylhexatriene and Laurdan anisotropy experiments showed that an S-linkage increased and NH- and CH(2)-linkages decreased the stability of PSM-analogue bilayer membranes as compared to PSM. When the polarity of the interface was probed using Laurdan, S-PSM appeared to have a lower polarity as compared to PSM whereas NH-PSM and CH(2)-PSM had higher polarities of their respective interfaces. Fluorescence quenching-studies with cholestatrienol showed that all compounds formed SM/cholesterol-rich domains. The S-PSM/cholesterol and PSM/cholesterol domains displayed a similar thermostability, whereas NH PSM/cholesterol and CH(2)-PSM/cholesterol domains were less thermostable. DSC on vesicles containing the PSM-analogues showed a more complex melting behavior as compared to PSM, whereas equimolar mixtures of the PSM-analogues and PSM showed almost ideal mixing with PSM for NH- and S-PSM. Our data show that the properties of the bond linking the phosphocholine head group to the 1-hydroxyl on the ceramide molecule is important for the stability of SM/SM and SM/cholesterol interactions. PMID- 18395515 TI - The alpha-5 helix of Bax is sensitive to ubiquitin-dependent degradation. AB - The pro-apoptotic protein Bax is instable in many cancer cells but the mechanism of Bax degradation remains unclear. Four different lengths of deductive Bax degradation sensitive (BDS) sequences within BH3-BH1 region, BDS-1 (Bax 67-124), BDS-3 (Bax 74-107), BDS-5 (Bax 67-107), and BDS-7 (Bax 74-124), were tested for the susceptibility to ubiquitin-dependent degradation. Both BDS-1 and BDS-7 which contain the alpha5 helix, a putative pore-forming domain of Bax, are sensitive to proteasome-dependent degradation and ubiquitin-conjugation. The Bax alpha5 deletion mutant (Bax-Deltaalpha5) was stable and also maintained its apoptosis inducing ability. Deletion of helices alpha1 and part of alpha2 (Bax-Delta1-66) or helices alpha3 and alpha4 (Bax-Deltaalpha3,4) did not affect the sensitivity to degradation. However, Bax-Delta1-66 mutant was not able to induce apoptosis. Thus, we propose that the alpha5 helix of Bax is sensitive to ubiquitin-dependent degradation. Moreover, Bax mutant retains its pro-apoptosis ability when the alpha5 helix is deleted. PMID- 18395516 TI - Single amino acid of an octopamine receptor as a molecular switch for distinct G protein couplings. AB - Octopamine (OA) is thought to be the invertebrate counterpart of noradrenaline and regulates various behavioral patterns of invertebrates by activating OA receptors. As a typical G protein-coupled receptor, BmOAR1, a Bombyx mori alpha adrenergic-like OA receptor, is coupled to both G(s) and G(q) proteins to induce the release of the intracellular second messengers cAMP and Ca(2+). In this study, we examined the pharmacological and functional properties of the cloned OA receptor, using OA enantiomers. The wild-type OA receptor exhibited significant stereoselectivity for OA enantiomers in cAMP production and binding affinity, but not in calcium signaling response. On the contrary, the Y412F mutant abolished the discrimination between OA enantiomers in the binding affinity and did not evoke any cAMP signaling response. This mutant exhibited levels of potency and efficacy similar to those of the wild-type receptor in the calcium assays. Taken together, these results suggest that Tyr412 might act as a molecular switch to regulate distinct G protein couplings, and a sequential activation model is proposed for such specific-residue-dependent, selective activation in receptors that are coupled to multiple G proteins. PMID- 18395517 TI - Induction of MHC class I-related chain B (MICB) by 5-aza-2'-deoxycytidine. AB - 5-Aza-2'-deoxycytidine (5-aza-dC), a DNA methyltransferase inhibitor, exerts antitumor activity through induction of cell cycle arrest, apoptosis and DNA damage. In this study, we showed that MHC class I-related chain B (MICB), a ligand of the NKG2D receptor expressed by natural killer cells and activated CD8(+) T cells, was upregulated following 5-aza-dC treatment. The upregulation of MICB was accompanied by promoter DNA demethylation and DNA damage. Furthermore, the upregulation of MICB was partially prevented by pharmacological or genetic inhibition of ataxia telangiectasia mutated (ATM) kinase. Our results suggest that promoter DNA demethylation, in combination with DNA damage, contribute to the upregulation of MICB induced by 5-aza-dC. PMID- 18395518 TI - Driving force of binding of amyloid beta-protein to lipid bilayers. AB - Amyloid beta-protein (Abeta) has been reported to interact with a variety of lipid species, although the thermodynamic driving force remains unclear. We investigated the binding of Abetas labeled with the dye diethylaminocoumarin (DAC Abetas) to lipid bilayers under various conditions. DAC-Abeta-(1-40) electrostatically bound to anionic and cationic lipids at acidic and alkaline interfacial pH, respectively. However, at neutral pH, electroneutral Abeta did not bind to these lipids, indicating little hydrophobic interaction between Abeta (1-40) and the acyl chains of lipids. In contrast, DAC-Abeta associated with glycolipids even under electroneutral conditions. These results suggested that hydrogen-bonding as well as hydrophobic interactions with sugar groups of glycolipids drive the membrane binding of Abeta-(1-40). PMID- 18395519 TI - Soluble tissue factor has unique angiogenic activities that selectively promote migration and differentiation but not proliferation of endothelial cells. AB - The level of circulating tissue factor (TF) is up-regulated in human angiogenesis related malignancies. However, whether circulating TF has angiogenic activities has not been determined. Soluble TF (sTF) is the main domain of circulating TF. Here, using cell migration, wound healing, and tubule formation assays, human recombinant sTF was found to significantly promote the migration and differentiation of endothelial cells. The stress fiber formation and rearrangement induced by sTF observed through immunofluorescence microscope may be responsible for the stimulatory migration effect of sTF. Nevertheless, sTF had no effects on endothelial cell proliferation. Interestingly, sTF can be internalized by endothelial cells, which implies a novel mechanism for sTF in angiogenesis. These results suggest that sTF has unique angiogenic activities and may serve as a potential therapeutic target to treat diseases associated with angiogenesis such as cancer and rheumatoid arthritis. PMID- 18395520 TI - A novel protein, MUDENG, induces cell death in cytotoxic T cells. AB - A screening system comprised of a randomized hybrid-ribozyme library has previously been used to identify pro-death genes in Fas-mediated apoptosis, and short sequence information of candidate genes from this system was previously reported by Kawasaki and Taira [H. Kawasaki, K. Taira, A functional gene discovery in the Fas-mediated pathway to apoptosis by analysis of transiently expressed randomized hybrid-ribozyme libraries, Nucleic Acids Res. 30 (2002) 3609 3614]. In this study, we have cloned the full-length of the candidate's open reading frames and found that one of the candidates, referred to as MUDENG (Mu-2 related death-inducing gene), which is composed of 490 amino acids that contain the adaptin domain found in the mu2 subunit of APs related to clathrin-mediated endocytosis, is able to induce cell death by itself. Ectopic expression of MUDENG induced cell death in Jurkat T cells and HeLa cells. In addition, when MUDENG expression was evaluated by immnuohistochemical staining, it was found in most tissues, including the intestine and testis. Furthermore, MUDENG appears to be evolutionary conserved from mammals to amphibians, suggesting that it may have a common role in cell death. Taken together, these results suggest that MUDENG is likely to play an important role in cell death in various tissues. PMID- 18395521 TI - Autoproteolytic stability of a trypsin from the marine crab Cancer pagurus. AB - Autoproteolytic stability is a crucial factor for the application of proteases in biotechnology. In contrast to vertebrate enzymes, trypsins from shrimp and crayfish are known to be resistant against autolysis. We show by characterisation of a novel trypsin from the gastric fluid of the marine crab Cancer pagurus that this property might be assigned to the entire class of crustaceans. The isolated and cloned crab trypsin (C.p.TryIII) exhibits all characteristic properties of crustacean trypsins. However, its overall sequence identity to other trypsins of this systematic class is comparatively low. The high resistance against autoproteolysis was determined by mass spectrometry, which revealed a low susceptibility of the N-terminal domain towards autolysis. By homology modelling of the tertiary structure, the elevated stability was attributed to the distinctly different pattern of autolytic cleavage sites, which is conserved in all known crustacean trypsin sequences. PMID- 18395522 TI - Interleukin-6 serum level assessment using a new qualitative point-of-care test in sepsis: A comparison with ELISA measurements. AB - OBJECTIVES: Interleukin-6 (IL-6) is involved in inflammatory diseases, provides prognostical information, and allows the early identification and monitoring of septic patients. We investigated whether IL-6 can be assessed using a new densitometric point-of-care (POC) bedside assay. DESIGN AND METHODS: 392 samples were prospectively collected from 57 intensive care unit patients (38 male, age: 45.2 +/- 16.9years, APACHE II score: 25.4 +/- 4.8). Blinded IL-6 measurements were performed using conventional semiautomatic enzyme-linked immunosorbent assays (ELISA) and the POC test. RESULTS: Mean IL-6 levels were 102.9 +/- 388.6pg/mL (ELISA) and 97.0 +/- 535.5 (POC). A significant correlation was found (p < 0.0001, r = 0.92). The sensitivity/specificity for sepsis was 82.6%/86.5% (ELISA, AUC: 0.881), and 76.4%/95.0% (POC, AUC: 0.868). CONCLUSIONS: Here we demonstrate significant correlations of IL-6 levels determined using a POC test and semiautomatic ELISA. ROC analyses revealed no significant differences between the two tests. With a turn-around time of 20min, the bedside IL-6 test is a new tool that may help to initiate early goal-directed therapy. PMID- 18395523 TI - The sodium pump Ena1p provides mechanistic insight into the salt sensitivity of vacuolar protein sorting mutants. AB - The vacuolar/endosomal network has an important but as yet undefined role in the cellular tolerance to salt stress. We hypothesized that the mechanistic basis for the importance of vacuolar protein sorting (vps) components in salt tolerance is the targeting of the crucial sodium exporter Ena1p to the plasma membrane. The link between Ena1p and the vps components was established by the observation that overexpression of Ena1p could suppress the salt sensitivity of the ESCRT knockouts vps20Delta, snf7/vps32Delta and snf8/vps22Delta. To further investigate this functional interaction, fluorescence microscopy was utilized to monitor localization of GFP-tagged Ena1p. For all analyzed vps mutants, Ena1p seemed properly localized to the plasma membrane, even during saline growth. However, quantitative differences in plasma membrane localized Ena1p were recorded; e.g. the highly salt sensitive pep12Delta mutant exhibited substantially enhanced Ena1p levels. In addition, the kinetics of Ena1p localization to the plasma membrane was severely delayed in several vps mutants, and this delay correlated to the salt specific growth defect. This paper discusses potential mechanistic hypotheses, like Ena1p transporter activity or localization kinetics, or ESCRT component's influence on signaling, for linking endosomal sorting functions to cellular salt sensitivity. PMID- 18395524 TI - ROS accumulation and oxidative damage to cell structures in Saccharomyces cerevisiae wine strains during fermentation of high-sugar-containing medium. AB - To further elucidate the impact of fermentative stress on Saccharomyces cerevisiae wine strains, we have here evaluated markers of oxidative stress, oxidative damage and antioxidant response in four oenological strains of S. cerevisiae, relating these to membrane integrity, ethanol production and cell viability during fermentation in high-sugar-containing medium. The cells were sampled at different fermentation stages and analysed by flow cytometry to evaluate membrane integrity and accumulation of reactive oxygen species (ROS). At the same time, catalase and superoxide dismutase activities, trehalose accumulation, and protein carbonylation and degradation were measured. The results indicate that the stress conditions occurring during hypoxic fermentation in high-sugar-containing medium result in the production of ROS and trigger an antioxidant response. This involves superoxide dismutase and trehalose for the protection of cell structures from oxidative damage, and protein catabolism for the removal of damaged proteins. Cell viability, membrane integrity and ethanol production depend on the extent of oxidative damage to cellular components. This is, in turn, related to the 'fitness' of each strain, which depends on the contribution of individual cells to ROS accumulation and scavenging. These findings highlight that the differences in individual cell resistances to ROS contribute to the persistence of wine strains during growth under unfavourable culture conditions, and they provide further insights into our understanding of yeast behaviour during industrial fermentation. PMID- 18395525 TI - Protein and lipid nitration: role in redox signaling and injury. AB - Protein and lipid nitration represent novel footprints of oxidative and nitrative stress processes. In this review, we first discuss the mechanisms of formation of protein 3-nitrotyrosine and nitrated fatty acids as well as their key biological and signaling actions. Elevation of protein 3-nitrotyrosine levels is associated to tissue injury, and some specific nitrated proteins play a causative role in disease progression; on the other hand, the substantiation on the role of tyrosine nitration on redox signaling is rather scarce. Herein, we also provide evidence to support that the nitration of lipids (i.e. to nitrofatty acids) results in the formation of novel endogenous modulators of redox processes, partially counteracting pro-inflammatory effects of oxidant exposure. PMID- 18395526 TI - Redox control in trypanosomatids, parasitic protozoa with trypanothione-based thiol metabolism. AB - Trypanosomes and leishmania, the causative agents of several tropical diseases, possess a unique redox metabolism which is based on trypanothione. The bis(glutathionyl)spermidine is the central thiol that delivers electrons for the synthesis of DNA precursors, the detoxification of hydroperoxides and other trypanothione-dependent pathways. Many of the reactions are mediated by tryparedoxin, a distant member of the thioredoxin protein family. Trypanothione is kept reduced by the parasite-specific flavoenzyme trypanothione reductase. Since glutathione reductases and thioredoxin reductases are missing, the reaction catalyzed by trypanothione reductase represents the only connection between the NADPH- and the thiol-based redox metabolisms. Thus, cellular thiol redox homeostasis is maintained by the biosynthesis and reduction of trypanothione. Nearly all proteins of the parasite-specific trypanothione metabolism have proved to be essential. PMID- 18395527 TI - 6-Hydroxydopamine (6-OHDA) induces Drp1-dependent mitochondrial fragmentation in SH-SY5Y cells. AB - Mitochondrial alterations have been associated with the cytotoxic effect of 6 hydroxydopamine (6-OHDA), a widely used neurotoxin to study Parkinson's disease. Herein we studied the potential effects of 6-OHDA on mitochondrial morphology in SH-SY5Y neuroblastoma cells. By immunofluorescence and time-lapse fluorescence microscopy we demonstrated that 6-OHDA induced profound mitochondrial fragmentation in SH-SY5Y cells, an event that was similar to mitochondrial fission induced by overexpression of Fis1p, a membrane adaptor for the dynamin related protein 1 (DLP1/Drp1). 6-OHDA failed to induce any changes in peroxisome morphology. Biochemical experiments revealed that 6-OHDA-induced mitochondrial fragmentation is an early event preceding the collapse of the mitochondrial membrane potential and cytochrome c release in SH-SY5Y cells. Silencing of DLP1/Drp1, which is involved in mitochondrial and peroxisomal fission, prevented 6-OHDA-induced fragmentation of mitochondria. Furthermore, in cells silenced for Drp1, 6-OHDA-induced cell death was reduced, indicating that a block in mitochondrial fission protects SH-SY5Y cells against 6-OHDA toxicity. Experiments in mouse embryonic fibroblasts deficient in Bax or p53 revealed that both proteins are not essential for 6-OHDA-induced mitochondrial fragmentation. Our data demonstrate for the first time an involvement of mitochondrial fragmentation and Drp1 function in 6-OHDA-induced apoptosis. PMID- 18395528 TI - Nitrated fatty acids: mechanisms of formation, chemical characterization, and biological properties. AB - Nitrated derivatives of unsaturated fatty acids are formed under oxidative and nitrative stress conditions, and are detected and structurally characterized in cell membranes, cardiac tissue, human plasma, and urine. Nitro-fatty acids display pleiotropic activities, including modulation of macrophage activation, prevention of leukocyte and platelet activation, and promotion of blood vessel relaxation. However, mechanisms of formation and levels reached in inflammatory milieu are poorly characterized. In this review, we discuss potential mechanisms of formation of nitro-fatty acids and their key chemical and biochemical properties. A major focus is to analyze nitrated lipids as novel signaling mediators leading to secondary changes in protein function via electrophilic based modifications as well as inhibition of inflammatory cell function, thus representing the convergence of lipid and nitric oxide signaling pathways. PMID- 18395530 TI - Molecular and kinetic characterization of two UDP-glucose pyrophosphorylases, products of distinct genes, from Arabidopsis. AB - UDP-glucose pyrophosphorylase (UGPase) is an important enzyme in the production (and conversions) of UDP-glucose, a key precursor for carbohydrate biosynthesis. cDNAs corresponding to two UGPase isozymes in Arabidopsis were overexpressed in Escherichia coli and, subsequently, the recombinant proteins were purified and characterized. Both proteins were highly conserved, sharing 93% identity. Based on crystal structure-derived images, the main amino acid differences mapped to N- and C-termini domains, but not to central active site region. The two proteins existed mainly as monomers, and they had similar molecular masses of ca. 53 kDa. However, comparison of molecular masses of UGPases from Arabidopsis root and leaf extracts revealed that the root protein was slightly larger, suggesting a post translational modification. Specific activity of the purified UGPase-1 was ca. 10 30% lower than that of UGPase-2, depending on direction of the reaction, whereas its K(m) values with all substrates in both directions of the reaction were consistently ca. twice lower than those of UGPase-2 (0.03-0.14 mM vs. 0.07-0.36 mM, respectively). Both proteins were "true" UGPases, and had no activity with ADP-glucose/ATP or galactose-1-P. Equilibrium constant for both proteins was ca. 0.3, suggesting preference for the pyrophosphorolysis direction of the reaction. The data are discussed with respect to potential roles of UGPase in carbohydrate synthesis/metabolism in Arabidopsis. PMID- 18395529 TI - The functional interaction on in vitro gene expression of APOA5 SNPs, defining haplotype APOA52, and their paradoxical association with plasma triglyceride but not plasma apoAV levels. AB - Plasma triglyceride (TG) and apoAV levels are reported to be positively correlated, yet SNPs defining haplotype APOA52 have consistently shown association with elevated plasma triglyceride (TG) but not plasma apoAV levels. We previously reported that individually -1131T>C, -3A>G and +1891T>C did not influence luciferase activity or in vitro translation efficiency. To investigate the combined effect of these SNPs additional constructs were examined. Compared to the wildtype -1131T/-3A/+1891T (TAT), the triple rare allele construct -1131C/ 3G/+1891C (CGC) conferred 46% lower luciferase activity (p<0.0001), showing these SNPs are acting co-operatively. Although only these two combinations occur in vivo, we experimentally altered the TAT construct one site at a time; -3G (TGT) had the largest effect (94% lower luciferase), with lesser effects from CAT ( 77%) and TAC (-70.3%) (all p<0.0001). Deletion constructs excluding one site at a time showed that -3G/1891C ( -GC) in combination, compared to -AT, was having the largest effect on luciferase activity (-59%, p=0.055). Using sequence homology and EMSA analysis no transcription factor binding at -1131 or +1891 was identified, though +1891 lies within a putative mRNA stability motif. Taken together, these data identify -3A>G in the Kozak sequence as functional, affecting translation initiation and driving the haplotype effects, while showing interaction with +1891T>C and to a lesser extent -1131T>C. A paradox arises since these results predict that APOA52 will lead to reduced apoAV with concomitant reduced LPL activation or lipoprotein-receptor interaction, resulting in higher plasma TG levels. We conclude that APOA5 expression, and not circulating plasma apoAV levels, is causatively associated with plasma TG levels. PMID- 18395531 TI - Formation of amyloid fibrils by bovine carbonic anhydrase. AB - Amyloids are typically characterized by extensive aggregation of proteins where the participating polypeptides are involved in formation of intermolecular cross beta-sheet structures. Alternate structure attainment and amyloid formation has been hypothesized to be a generic property of a polypeptide, the propensities of which vary widely depending on the polypeptide involved and the physicochemical conditions it encounters. Many proteins that exist in the normal form in-vivo have been shown to form amyloid when incubated in partially denaturing conditions. The protein bovine carbonic anhydrase II (BCA II) when incubated in mildly denaturing conditions showed that the partially unfolded conformers assemble together and form ordered amyloid aggregates. The properties of these aggregates were tested using the traditional Congo-Red (CR) and Thioflavin-T (ThT) assays along with fluorescence microscopy, transmission electron microscopy (TEM), and circular dichroism (CD) spectroscopy. The aggregates were found to possess most of the characteristics ascribed to amyloid fibers. Thus, we report here that the single-domain globular protein, BCA II, is capable of forming amyloid fibrils. The primary sequence of BCA II was also analyzed using recurrence quantification analysis in order to suggest the probable residues responsible for amyloid formation. PMID- 18395532 TI - Serotonin, stress, and conditioning. PMID- 18395533 TI - Fractures related to occlusal overload with single posterior implants: a clinical report. AB - This clinical report describes 2 patient situations in which fractures related to occlusal overload occurred with a single posterior implants. The initial clinical presentation of both patients appeared to be screw loosening, but upon further examination, implant and abutment fractures were identified. Several factors are described that have been implicated in the etiology of implant fractures, including occlusal overload, implant location, inadequate fit of the prosthesis, design of the prosthesis, progressive bone loss, metal fatigue, implant diameter, manufacturing defects, and galvanic activity. This article describes the management of implant and abutment fractures and discusses possible mechanisms of failure for the patient situations presented. Careful treatment planning and execution of implant therapy is necessary to minimize the risk of implant and component fractures. PMID- 18395534 TI - Oral rehabilitation of a 12-year-old patient diagnosed with a central giant cell granuloma using a fibula graft and an implant-supported prosthesis: a clinical report. AB - This clinical report describes the oral rehabilitation of a 12-year-old boy with a central giant cell granuloma of the mandible. He underwent en bloc resection of a central giant cell granuloma, free vascularized fibula reconstruction, implant placement, and prosthesis fabrication. The multidisciplinary approach successfully restored function and esthetics. Considerations regarding the extensive surgical and prosthetic rehabilitation of a young adult with remaining growth are discussed. PMID- 18395535 TI - Removable partial denture design using milled abutment surfaces and minimal soft tissue coverage for periodontally compromised teeth: a clinical report. AB - This clinical report describes the treatment of a partially edentulous patient with periodontally compromised teeth using a combination of single crowns and a removable partial denture (RPD). The RPD was designed to combine the benefits of milled surfaces and hygienic principles while allowing modification and addition of artificial teeth, should natural teeth be lost. PMID- 18395536 TI - Residual dentin thickness in bifurcated maxillary first premolars after root canal and post space preparation with parallel-sided drills. AB - STATEMENT OF PROBLEM: The primary factor that determines the prognosis of endodontically treated and restored teeth is preservation of sound dentin. The potential of these teeth to fracture is directly related to the amount of dentin removed. PURPOSE: The purpose of this study was to measure residual dentin thickness of bifurcated maxillary first premolars, in vitro, after root canal and conservative post space preparation. MATERIAL AND METHODS: Human premolars (n=13) with bifurcation at the junction of cervical and middle root thirds were embedded in a muffle device and horizontally sectioned 2, 4, and 6 mm apically to the cement-enamel junction. Residual dentin thickness was measured at the buccal, lingual, mesial, and distal aspects, apically to the bifurcation, and at the buccal or lingual, mesial, and distal aspects, coronally to the bifurcation. This procedure was repeated at baseline, after endodontic preparation (K-40 file), and after preparing the post space (ParaPost drills Nos. 3 and 4). Residual dentin thickness was analyzed using 3- or 4-way repeated measures ANOVA (alpha=.05). RESULTS: Endodontic therapy and post preparation reduced more dentin in the bifurcation aspects of both roots compared to outer aspects (P=.001). Post space preparation jeopardized the roots due to a residual dentin thickness of less than the recommended minimum 1-mm thickness in 61% (lingual) and 77% (buccal) of roots. CONCLUSIONS: Lack of residual dentin thickness after post preparation implies that the use of posts in maxillary first premolars should be limited. When mandatory, it is recommended that the lingual root be used and the buccal root be avoided. PMID- 18395537 TI - Dimensional accuracy of resultant casts made by a monophase, one-step and two step, and a novel two-step putty/light-body impression technique: an in vitro study. AB - STATEMENT OF PROBLEM: Dimensional accuracy when making impressions is crucial to the quality of fixed prosthodontic treatment, and the impression technique is a critical factor affecting this accuracy. PURPOSE: The purpose of this in vitro study was to compare the dimensional accuracy of a monophase, 1- and 2-step putty/light-body, and a novel 2-step injection impression technique. MATERIAL AND METHODS: A stainless steel model with 2 abutment preparations was fabricated, and impressions were made 15 times with each technique. All impressions were made with an addition-reaction silicone impression material (Aquasil) and a stock perforated metal tray. The monophase impressions were made with regular body material. The 1-step putty/light-body impressions were made with simultaneous use of putty and light-body materials. The 2-step putty/light-body impressions were made with 2-mm-thick resin-prefabricated copings. The 2-step injection impressions were made with simultaneous use of putty and light-body materials. In this injection technique, after removing the preliminary impression, a hole was made through the polymerized material at each abutment edge, to coincide with holes present in the stock trays. Extra-light-body material was then added to the preliminary impression and further injected through the hole after reinsertion of the preliminary impression on the stainless steel model. The accuracy of the 4 different impression techniques was assessed by measuring 3 dimensions (intra- and interabutment) (5-mum accuracy) on stone casts poured from the impressions of the stainless steel model. The data were analyzed by 1-way ANOVA and Student Newman-Keuls test (alpha=.05). RESULTS: The stone dies obtained with all the techniques had significantly larger dimensions as compared to those of the stainless steel model (P<.01). The order for highest to lowest deviation from the stainless steel model was: monophase, 1-step putty/light body, 2-step putty/light body, and 2-step injection. Significant differences among all of the groups for both absolute dimensions of the stone dies, and their percent deviations from the stainless steel model (P<.01), were noted. CONCLUSIONS: The 2-step putty/light body and 2-step injection techniques were the most dimensionally accurate impression methods in terms of resultant casts. PMID- 18395538 TI - The effect of different investment techniques on the surface roughness and irregularities of gold palladium alloy castings. AB - STATEMENT OF PROBLEM: Surface roughness and irregularities can affect the fit of a restoration. It is unknown whether different investment techniques have an effect on surface roughness and irregularities of gold palladium alloy castings. PURPOSE: The purpose of this study was to evaluate the surface roughness and irregularities of gold palladium alloy castings obtained using different investment techniques. MATERIAL AND METHODS: Forty disk-shaped wax patterns were prepared and divided into 4 groups. A phosphate-bonded, carbon-free investment was used for the investment procedures. Twenty specimens were invested using a vacuum mixer, while the remainder were invested using a vacuum mixer and investor. The specimens in both broad categories were divided evenly to set in 2 different conditions of pressure. Half were left to set under atmospheric pressure for 1 hour, while the rest were placed in a compression chamber under a pressure of 3 bars for 24 minutes, then allowed to bench set for another 36 minutes. A profilometer was used to evaluate the surface roughness (mum) of the castings. The specimens were also studied under x10 magnification for surface irregularities. Two-way ANOVA was used to examine the relationships among surface roughness, investing, and pressure (alpha=.05). Logistic regression was used to explore the relationships among surface irregularities, investing, and pressure. RESULTS: Two-way ANOVA for surface roughness did not reveal any statistically significant differences. However, there was a trend for the specimens set under atmospheric pressure to present lower values of surface roughness than the specimens set under positive pressure (P=.095). The logistic regression for surface irregularities showed a highly significant impact of pressure. The odds of any surface irregularities for specimens under atmospheric pressure were 9.12 times higher than the odds for specimens under positive pressure (P=.016). CONCLUSIONS: Within the limitations of this study, the results suggest that specimens set under atmospheric pressure are much more likely to present surface irregularities than specimens set under positive pressure. The use of pressure can help produce castings with fewer surface irregularities. PMID- 18395539 TI - Effect of cement selection and finishing technique on marginal opening of cast gold inlays. AB - STATEMENT OF PROBLEM: The standard cement for partial-veneer cast gold restorations has been zinc phosphate. With increasing interest in using resin and resin-modified glass ionomer cements for this purpose, marginal adaptation with use of these newer cements must be assessed. PURPOSE: The purpose of this in vitro study was to determine if the marginal adaptation of cast gold inlays is comparable for 3 different luting agents and 3 finishing techniques. MATERIAL AND METHODS: Cast gold inlays were cemented into triangular preparations on the buccal surface of 45 extracted human molars. Three operators each used a different technique for finishing margins: finishing during cementation (FD), finishing before cementation (FB), and finishing before and during cementation (FBA). The 15 teeth for each operator were divided into 3 subgroups of 5 each to accommodate 3 cements: zinc phosphate (Fleck's zinc phosphate: ZP), resin modified glass ionomer (RelyX Luting: RXL), and self-adhesive modified resin (RelyX Unicem: RXU). For all specimens, fine sandpaper disks followed by fine cuttle disks were used for finishing the castings. The marginal gap was measured using a Nikon measuring microscope at x50 magnification. Data were analyzed with a 1-way ANOVA for each cement (alpha=.05). RESULTS: There were significant differences among finishing techniques for each cement (P<.05). For ZP, the smallest mean marginal gaps were for FD (31 microm) and FBA (42 microm). For RXL, FBA produced the smallest gap (19 microm). For RXU, FB (23 microm) and FBA (22 microm) were optimal. CONCLUSIONS: Within the limitations of this in vitro study, the marginal gap attained with cementation of partial-veneer cast gold restorations with ZP (31 microm) can also be obtained using RXL or RXU. The techniques of FD or FBA are best used for ZP, whereas the FBA technique is best for RXL, and either FB or FBA is optimal for RXU. The FBA technique yielded a small gap; this was common to all 3 cements. PMID- 18395540 TI - Classification system for conventional crown and fixed partial denture failures. AB - The dental literature is replete with reports on the many aspects of failure encountered with traditional fixed prosthodontic treatment, including longitudinal survival studies of crowns and fixed partial dentures and reasons for failures. However, criteria for grading or classifying the type and severity of these failures are inadequate. A classification system for conventional fixed prosthodontic failures based on severity is presented. PMID- 18395541 TI - Occlusion for fixed prosthodontics: a historical perspective of the gnathological influence. AB - This article addresses the historical perspective of the gnathological influence upon the concepts of occlusion for fixed prosthodontics. A critical assessment and subsequent scientific validation of occlusal theories require an understanding of their evolution in the formative years and the subsequent development of effective models for clinical practice. While gnathological concepts offer a structured methodology for prosthodontic procedures, further research is needed to corroborate current occlusal treatment approaches. This review focuses on the "classic" fixed prosthodontic literature and the currently available scientific literature involving fixed prosthodontic dentate occlusion and gnathology. A MEDLINE search was performed to identify English-language peer reviewed publications spanning the last 56 years, along with an extensive hand search for years prior. Electronic searches of the literature were performed in MEDLINE using the key words: case series, clinical trials, cohort studies, fixed partial denture occlusion, dental occlusion, dental occlusion research, centric relation, incisal guidance, maximal intercuspation, occlusal vertical dimension, and occlusion, in various combinations to obtain potential references for review. A total of 10,382 English-language nonduplicate titles were obtained for 1950 2006 for the key words "dental occlusion." Other key word searches produced smaller numbers of articles, many of which were duplicates due to multiple searches and were subsequently eliminated. Manual hand searching of the MEDLINE reference list and other journals of interest was performed to identify any articles missed in the original search. Articles were included for review if they contained emerging occlusal theories, new technologies, or occlusal studies that included multiple subjects in contrast to case reports. PMID- 18395542 TI - Ten-year follow-up of mandibular advancement devices for the management of snoring and sleep apnea. AB - STATEMENT OF PROBLEM: Intraoral mandibular advancement devices have become widely used in recent years for the management of snoring and sleep apnea, and short term effectiveness has been demonstrated. However, there is a shortage of data regarding long-term compliance. PURPOSE: The purpose of this study was to investigate the long-term compliance of patients who were provided with a mandibular advancement device. MATERIAL AND METHODS: Records of 180 patients who were provided with a mandibular advancement device in 1996 were available for review. A questionnaire was sent to all of these patients inquiring about continued device usage, comfort, and effectiveness. Questions were also asked about smoking, alcohol consumption, height, and weight. Data were analyzed with chi-square tests for any association between these factors and success of the device (alpha=.05). RESULTS: The response rate was 40%, with 72 replies. Of this number, 34 patients were currently wearing the device every night, with a further 13 wearing the device for up to 6 nights per week. Thirty-one of the respondents who were wearing the device felt more refreshed on waking. The median body mass index (BMI) was 30, 8 were smokers, and 12 subjects drank more than 20 units of alcohol per week. Few adverse effects of the device were reported. CONCLUSIONS: The mandibular advancement device appears to be an effective long-term solution for a significant number of patients with problem snoring and also those with mild to moderate obstructive sleep apnea. PMID- 18395543 TI - A technique for stabilizing casts during the mounting procedure. PMID- 18395544 TI - A modified spruing technique for multiple-unit fixed partial denture frameworks. PMID- 18395545 TI - A simple method to duplicate a denture for an implant surgical guide. PMID- 18395546 TI - Therapeutic approaches for control of transcription factors in allergic disease. AB - The inflammatory response observed in allergic disease involves multiple cell types but is orchestrated in part by the T(H)2 cytokines IL-4, IL-5, and IL-13. In recent years, the transcription factors that control the expression and function of these cytokines have been elucidated, including signal transducer and activator of transcription 6, GATA3, nuclear factor of activated T cells, and nuclear factor kappaB. These molecules are attractive targets for therapeutic intervention because they regulate the expression of numerous effector molecules and functions simultaneously. For instance, the immunosuppressive agents glucocorticoids and cyclosporin A both function by repressing the activity of transcription factors through a variety of mechanisms. In this review we examine the role of each transcription factor in allergic disease and discuss approaches that have been taken to therapeutically interfere with transcription factor function in allergic disease. PMID- 18395547 TI - T-cell effector pathways in allergic diseases: transcriptional mechanisms and therapeutic targets. AB - Originally interpreted within the framework of a binary T(H)1/T(H)2 paradigm, our knowledge of the pathogenesis of atopic diseases has broadened to incorporate the contribution of T regulatory cells and the newly described proinflammatory T(H)17 cell lineage. The commitment of peripheral T-cell clones to undergo differentiation into one of those lineages is shaped by self-reinforcing transcriptional circuitries that center on key transcriptional regulators: T-box expressed in T cells (T(H)1), GATA-3 (T(H)2), forkhead box p3 (T regulatory cells), and retinoid-related orphan receptor gammatau/retinoid-related orphan receptor alpha (T(H)17). These circuits function both to establish the respective lineage phenotype and to enable epigenetic changes that maintain those phenotypes long-term. This evolving view of how signaling and transcriptional networks generate effector T-cell responses suggests novel therapeutic approaches to reprogram effector T-cell lineage commitment in allergic diseases in favor of tolerance induction. PMID- 18395549 TI - Allergens are distributed into few protein families and possess a restricted number of biochemical functions. AB - BACKGROUND: Existing allergen databases classify their entries by source and route of exposure, thus lacking an evolutionary, structural, and functional classification of allergens. OBJECTIVE: We sought to build AllFam, a database of allergen families, and use it to extract common structural and functional properties of allergens. METHODS: Allergen data from the Allergome database and protein family definitions from the Pfam database were merged into AllFam, a database that is freely accessible on the Internet at http://www.meduniwien.ac.at/allergens/allfam/. A structural classification of allergens was established by matching Pfam families with families from the Structural Classification of Proteins database. Biochemical functions of allergens were extracted from the Gene Ontology Annotation database. RESULTS: Seven hundred seven allergens were classified by sequence into 134 AllFam families containing 184 Pfam domains (2% of 9318 Pfam families). A random set of 707 sequences with the same taxonomic distribution contained a significantly higher number of different Pfam domains (479 +/- 17). Classifying allergens by structure revealed that 5% of 3012 Structural Classification of Proteins families contained allergens. The biochemical functions of allergens most frequently found were limited to hydrolysis of proteins, polysaccharides, and lipids; binding of metal ions and lipids; storage; and cytoskeleton association. CONCLUSION: The small number of protein families that contain allergens and the narrow functional distribution of most allergens confirm the existence of yet unknown factors that render proteins allergenic. PMID- 18395550 TI - A polymorphism controlling ORMDL3 expression is associated with asthma that is poorly controlled by current medications. AB - BACKGROUND: The specific genetic contributions to childhood asthma have been difficult to elucidate. A recent whole-genome association study suggested that single nucleotide polymorphisms at loci controlling the expression of the ORMDL3 gene and others in the neighborhood of the NRG1 and ERO1LB genes might be important. OBJECTIVE: We sought to replicate the associations of these genetic markers with asthma in a large population of asthmatic patients from Scotland and to assess the effect of these variants on asthma outcomes. METHODS: Using mouthwash-derived DNA and clinical interviews and measurements, we investigated the association of 3 single nucleotide polymorphisms in the candidate genes with susceptibility to asthma in a case-control study and also exacerbations in a group of 1054 patients aged 3 to 22 years. RESULTS: A common C/T polymorphism at a locus controlling ORMDL3 gene expression (rs7216389) was significantly associated with the risk of childhood asthma (P = 1.73 x 10(-12)), with a single copy of the T allele conferring an odds ratio of 1.50 (95% CI, 1.24-1.81) and 2 copies of the T allele conferring an odds ratio of 2.11 (95% CI, 1.71-2.61), respectively. In asthmatic patients the T allele was associated with exacerbations of the condition (P = .008). Polymorphisms at the loci of nearby genes for NRG1 (rs4512342) and ERO1LB (rs10924993) were associated with neither the occurrence of nor exacerbations of asthma. CONCLUSION: A common genetic variation at a locus controlling the expression of the ORMDL3 locus increases the susceptibility to asthma and is associated with poor control of the condition in children and young adults. PMID- 18395551 TI - Corticosteroids enhance CD8+ T cell-mediated airway hyperresponsiveness and allergic inflammation by upregulating leukotriene B4 receptor 1. AB - BACKGROUND: Leukotriene B4 (LTB4) is a potent inflammatory lipid mediator that binds to LTB4 receptor 1 (BLT1). Ligation of BLT1 by LTB4 plays an important role in the recruitment of effector memory CD8+ T cells into the airways of sensitized and challenged mice. OBJECTIVES: The effects of the corticosteroid dexamethasone (DEX) on BLT1-expressing effector memory CD8+ T cells and effector memory CD8+ T cell-mediated airway hyperresponsiveness (AHR) and allergic inflammation were determined. METHODS: Effector memory CD8+ T cells were generated from ovalbumin(257-264)-primed mononuclear cells from OT-1 mice in the presence of IL 2. In some cultures DEX was added. The effects of DEX on BLT1 expression, LTB4 induced Ca2+ influx, phosphorylation of extracellular signal-regulated kinase 1/2, chemotaxis, and effector memory CD8+ T cell-mediated AHR were examined. RESULTS: DEX-treated effector memory CD8+ T cells showed significant increases in surface expression of BLT1, LTB4-induced intracellular Ca2+ influx, phosphorylation of extracellular signal-regulated kinase 1/2, and chemotaxis. Upregulation of BLT1 by DEX was accompanied by increased IL-2 receptor expression. Adoptive transfer of DEX-treated effector memory CD8+ T cells into ovalbumin-sensitized and ovalbumin-challenged CD8-/- mice resulted in significant increases in AHR, allergic inflammation, goblet cell metaplasia, and numbers of both CD8+ and CD4+ T cells in the bronchoalveolar lavage fluid and lungs. CONCLUSIONS: Corticosteroids upregulate BLT1 on effector memory CD8+ T cells and related signaling pathways and potentiate allergic airway inflammation and AHR induced by these cells. PMID- 18395552 TI - Cell-specific activation profile of extracellular signal-regulated kinase 1/2, Jun N-terminal kinase, and p38 mitogen-activated protein kinases in asthmatic airways. AB - BACKGROUND: Many airway cells manifest signs of chronic activation in asthma. The mechanism of this chronic activation is unknown. OBJECTIVES: We sought to study the activation of the mitogen-activated protein kinase (MAPK) signaling pathway in airway cells. METHODS: Endobronchial biopsy specimens from patients with severe and mild asthma (n = 17 in each group) and healthy control subjects (n = 15) were analyzed for the phosphorylated MAPKs extracellular signal-regulated kinase (ERK) 1/2, p38, and Jun N-terminal kinase (JNK) and their downstream effectors by means of immunofluorescence staining. Airway epithelial activation of ERK1/2 and p38 was studied by using Western blotting. Epithelial function was studied by means of real-time PCR, ELISA, and the thymidine incorporation assay. RESULTS: We detected strong phospho-ERK1/2 staining in airway epithelium and smooth muscle cells in biopsy specimens from asthmatic patients. Fluorescent areas per image, as well as mean fluorescence intensity, were significantly (P < .0001) different among the 3 study groups (patients with severe asthma, patients with mild asthma, and healthy control subjects). Patients with severe asthma also demonstrated strong phospho-p38 staining, mostly in epithelial cells, which was significantly different from that in patients with mild asthma (P = .0001) and healthy control subjects (P = .02). Phospho-JNK primarily stained airway smooth muscle cells. Healthy subjects showed the highest intensity of phospho-JNK staining compared with that seen in patients with severe (P = .004) and mild asthma (P = .003). Inhibition of ERK1/2 and p38 in primary airway epithelial cells blocked their proliferation and expression of select, but not all, chemokines. CONCLUSIONS: Significant phosphorylation of ERK1/2 and p38 and their correlation with disease severity suggests that the foregoing signaling pathways play an important role in asthma. The ERK1/2 and p38 pathways regulate epithelial cell secretory function and proliferation. PMID- 18395553 TI - Potential role of vasomotor effects of fibrinogen in bradykinin-induced angioedema. AB - BACKGROUND: Although bradykinin is known to play a major role in the pathophysiology of hereditary and angiotensin-converting enzyme inhibitor (ACEi) induced angioedema, other factors acting as triggers or enhancers are likely important as well. OBJECTIVE: We hypothesized that fibrinogen might contribute to ACEi-induced angioedema (eg, through direct actions on vascular tone). METHODS: Plasma levels of fibrinogen were determined in 59 patients with acute angioedema. Vascular activity of human and bovine fibrinogen and its effects on bradykinin induced vasodilation and phosphorylation of vasodilator-stimulated phosphoprotein were investigated in small (0.8-1.4 mm in diameter) porcine coronary artery and human internal thoracic artery (ITA) segments. RESULTS: In patients with ACEi induced angioedema, fibrinogen levels (481 +/- 22 mg/dL, n = 39) were significantly higher than in patients with idiopathic angioedema (302 +/- 15 mg/dL, P < .001). Fibrinogen (1-15 mumol/L) induced a concentration-dependent vasodilation in preconstricted small porcine coronary arteries (n = 13), reaching a maximum vasodilator effect of 70% +/- 4.7%. Likewise, fibrinogen induced a 52.1% +/- 9.1% (n = 7) vasodilation in ITA rings. Fibrinogen vasorelaxations were completely inhibited by abciximab and diminished by endothelial denudation and treatment with the nitric oxide synthase inhibitor L-nitroargininemethylester and glibenclamide (P < .01). Importantly, fibrinogen increased the vasodilator potency of bradykinin by 10-fold (P < .0001) and increased bradykinin-induced vasodilator-stimulated phosphoprotein phosphorylation (P < .01). CONCLUSION: The increase of plasma fibrinogen levels, its vasodilator activity in human ITAs, and the potentiation of bradykinin-induced vasodilation suggest that fibrinogen might contribute to the pathophysiology of ACEi-induced angioedema. Thus acute-phase proteins, such as fibrinogen, might be viewed as risk factors for bradykinin induced angioedema. PMID- 18395555 TI - The effects of laparoscopic Nissen fundoplication to enhance pulmonary function in the treatment of a patient with severe asthma and gastroesophageal reflux disease. PMID- 18395556 TI - Is exposure to media violence a public-health risk? PMID- 18395554 TI - Cat ownership is a risk factor for the development of anti-cat IgE but not current wheeze at age 5 years in an inner-city cohort. AB - BACKGROUND: Cat ownership is inversely associated with atopy and asthma in some areas of the world, but the relevance of cat ownership to allergic disease in the inner city is less known. OBJECTIVE: We sought to evaluate the relationship between cat ownership and the development of early sensitization and wheeze. METHODS: By using a prospective birth cohort study, Dominican and African American mothers living in New York City underwent repeated questionnaires about their child from birth to age 5 years. Sera collected from children at ages 2 (n = 323), 3 (n = 336), and 5 (n = 242) years were assayed for anti-cat IgE and anti Fel d 1 IgG antibodies. RESULTS: Cat ownership was a significant risk factor for the development of anti-cat IgE by age 2 years (risk ratio [RR], 6.4; 95% CI, 1.9 22) but not for anti-cat IgE development between the ages of 2 and 5 years (RR, 0.88; 95% CI, 0.24-2.3). Current wheeze was significantly more common among those children with anti-cat IgE at ages 3 (RR, 3.5; 95% CI, 2.1-6.0) and 5 (RR, 3.4; 95% CI, 2.3-4.9) years. Cat ownership was inversely associated with current wheeze at age 5 years among children without anti-cat IgE (RR, 0.26; 95% CI, 0.083-0.81). Among children with anti-cat IgE, a similar trend was observed (RR, 0.57; P = .044, Fisher exact test), although one with borderline statistical significance. CONCLUSIONS: Despite a positive association with sensitization, cat ownership in this inner-city cohort was inversely associated with wheeze, potentially suggesting an IgE-independent protective mechanism in this community. PMID- 18395557 TI - A global dialogue on a global crisis. PMID- 18395558 TI - Sale of infant cough medicines splutters to a halt. PMID- 18395559 TI - A new initiative and invitation for health monitoring, tracking, and evaluation. PMID- 18395560 TI - Intimate-partner violence and women's health. PMID- 18395561 TI - Screening for neuroblastoma: a resurrected idea? PMID- 18395562 TI - Human rotavirus vaccines: too early for the strain to tell. PMID- 18395564 TI - Fine-needle aspiration in rural areas. PMID- 18395563 TI - The Lancet-UCL Commission: health effects of climate change. PMID- 18395565 TI - Doctors' standard of proof rejected by lawyers? PMID- 18395566 TI - EQUATOR: reporting guidelines for health research. PMID- 18395567 TI - Claudia Garcia-Moreno. PMID- 18395568 TI - Symptoms. PMID- 18395569 TI - Cholesterol, statins, and mortality. PMID- 18395570 TI - Cholesterol, statins, and mortality. PMID- 18395571 TI - Cholesterol, statins, and mortality. PMID- 18395573 TI - Cholesterol, statins, and mortality. PMID- 18395574 TI - A wearable haemodialysis device. PMID- 18395575 TI - A wearable haemodialysis device. PMID- 18395577 TI - Intimate partner violence and women's physical and mental health in the WHO multi country study on women's health and domestic violence: an observational study. AB - BACKGROUND: This article summarises findings from ten countries from the WHO multi-country study on women's health and domestic violence against women. METHODS: Standardised population-based surveys were done between 2000 and 2003. Women aged 15-49 years were interviewed about their experiences of physically and sexually violent acts by a current or former intimate male partner, and about selected symptoms associated with physical and mental health. The women reporting physical violence by a partner were asked about injuries that resulted from this type of violence. FINDINGS: 24,097 women completed interviews. Pooled analysis of all sites found significant associations between lifetime experiences of partner violence and self-reported poor health (odds ratio 1.6 [95% CI 1.5-1.8]), and with specific health problems in the previous 4 weeks: difficulty walking (1.6 [1.5-1.8]), difficulty with daily activities (1.6 [1.5-1.8]), pain (1.6 [1.5 1.7]), memory loss (1.8 [1.6-2.0]), dizziness (1.7 [1.6-1.8]), and vaginal discharge (1.8 [1.7-2.0]). For all settings combined, women who reported partner violence at least once in their life reported significantly more emotional distress, suicidal thoughts (2.9 [2.7-3.2]), and suicidal attempts (3.8 [3.3 4.5]), than non-abused women. These significant associations were maintained in almost all of the sites. Between 19% and 55% of women who had ever been physically abused by their partner were ever injured. INTERPRETATION: In addition to being a breach of human rights, intimate partner violence is associated with serious public-health consequences that should be addressed in national and global health policies and programmes. PMID- 18395578 TI - Effectiveness of screening for neuroblastoma at 6 months of age: a retrospective population-based cohort study. AB - BACKGROUND: In Japan, a nationwide programme between 1984 and 2003 screened all infants for urinary catecholamine metabolites as a marker for neuroblastoma. Before 1989, this was done by qualitative spot tests for vanillylmandelic acid in urine, and subsequently by quantitative assay with high-performance liquid chromatography (HPLC). However, the Japanese government stopped the mass screening programme in 2003, after reports that it did not reduce mortality due to neuroblastoma. We aimed to assess the effectiveness of the programme, by comparing the rates of incidence and mortality from neuroblastomas diagnosed before 6 years of age in three cohorts. METHODS: We did a retrospective population-based cohort study on all children born between 1980 and 1998, except for a 2-year period from 1984. We divided these 22,289,695 children into three cohorts: children born before screening in 1980-83 (n=6,130,423); those born during qualitative screening in 1986-89 (n=5,290,412); and those born during quantitative screening 1990-98 (n=10,868,860). We used databases from hospitals, screening centres, and national cancer registries. Cases of neuroblastoma were followed up for a mean of 78.7 months. FINDINGS: 21.56 cases of neuroblastoma per 100,000 births over 72 months were identified in the qualitatively screened group (relative risk [RR] 1.87, 95% CI 1.66-2.10), and 29.80 cases per 100,000 births over 72 months in the quantitatively screened group (RR 2.58, 2.33-2.86). The cumulative incidence of neuroblastoma in the prescreening cohort (11.56 cases per 100,000 births over 72 months) was lower than that in other cohorts (p<0.0001 for all comparisons), but more neuroblastomas were diagnosed after 24 months of age in this cohort (p=0.0002 for qualitative screening vs prescreening, p<0.0001 for quantitative screening vs prescreening). Cumulative mortality was lower in the qualitative screening (3.90 cases per 100,000 livebirths over 72 months) and quantitative screening cohorts (2.83 cases) than in the prescreening cohort (5.38 cases). Compared with the prescreening cohort, the relative risk of mortality was 0.73 (95% CI 0.58-0.90) for qualitative screening, and 0.53 (0.42-0.63) for quantitative screening. Mortality rates for both the qualitative and quantitative screening groups were lower than were those for the prescreening cohort (p=0.0041 for prescreening vs qualitative screening, p<0.0001 for prescreening vs quantitative screening). INTERPRETATION: More infantile neuroblastomas were recorded in children who were screened for neuroblastoma at 6 months of age than in those who were not. The mortality rate from neuroblastoma in children who were screened at 6 months was lower than that in the prescreening cohort, especially in children screened by quantitative HPLC. Any new screening programme should aim to decrease mortality, but also to minimise overdiagnosis of tumours with favourable prognoses (eg, by screening children at 18 months). PMID- 18395580 TI - Red spots on the hands and red blood in the stools. PMID- 18395579 TI - Efficacy and safety of an oral live attenuated human rotavirus vaccine against rotavirus gastroenteritis during the first 2 years of life in Latin American infants: a randomised, double-blind, placebo-controlled phase III study. AB - BACKGROUND: Peak incidence of rotavirus gastroenteritis is seen in infants between 6 and 24 months of age. We therefore aimed to assess the 2-year efficacy and safety of an oral live attenuated human rotavirus vaccine for prevention of severe gastroenteritis in infants. METHODS: 15 183 healthy infants aged 6-13 weeks from ten Latin American countries randomly assigned in a 1 to 1 ratio to receive two oral doses of RIX4414 or placebo at about 2 and 4 months of age in a double-blind, placebo-controlled phase III study were followed up until about 2 years of age. Primary endpoint was vaccine efficacy from 2 weeks after dose two until 1 year of age. Treatment allocation was concealed from investigators and parents of participating infants. Efficacy follow-up for gastroenteritis episodes was undertaken from 2 weeks after dose two until about 2 years of age. Analysis was according to protocol. This study is registered with ClinicalTrials.gov, number NCT00140673 (eTrack444563-023). FINDINGS: 897 infants were excluded from the according-to-protocol analysis. Fewer cases (p<0.0001) of severe rotavirus gastroenteritis were recorded for the combined 2-year period in the RIX4414 group (32 [0.4%] of 7205; 95% CI 0.3-0.6) than in the placebo group (161 [2.3%] of 7081; 1.9-2.6), resulting in a vaccine efficacy of 80.5% (71.3-87.1) to 82.1% (64.6-91.9) against wild-type G1, 77.5% (64.7-86.2) against pooled non-G1 strains, and 80.5% (67.9-88.8) against pooled non-G1 P[8] strains. Vaccine efficacy for hospital admission for rotavirus gastroenteritis was 83.0% (73.1 89.7) and for admission for diarrhoea of any cause was 39.3% (29.1-48.1). No cases of intussusception were reported during the second year of follow-up. INTERPRETATION: Two doses of RIX4414 were effective against severe rotavirus gastroenteritis during the first 2 years of life in a Latin American setting. Inclusion of RIX4414 in routine paediatric immunisations should reduce the burden of rotavirus gastroenteritis worldwide. PMID- 18395581 TI - Health metrics and evaluation: strengthening the science. AB - With the growing importance of health in the global agenda comes the responsibility to develop a scientific foundation of metrics and evaluation. The scope of this emerging field can be viewed in terms of key topics, including health outcomes, other social outcomes related to health systems, health services, resource inputs, evaluations of programmes and systems, and analyses to support policy choice. It can also be defined in terms of key activities that are needed to strengthen the scientific basis of the field: development of new methods, instruments, software, and hardware; setting global norms and standards for data collection; increasing the availability of high-quality primary data; systematic analysis and synthesis of existing datasets; strengthening national capacity to obtain, analyse, and use data; and reporting and disseminating results. We explore in depth topics with major scientific challenges and institutional and cultural barriers that are slowing the development of the field. Cutting across the various topical areas and disciplinary approaches to these problems are some common scientific issues, including limited comparability of measurement, uncorrected known biases in data, no standard approach to missing data, unrealistic uncertainty estimates, and the use of disease models that have not been properly validated. Only through concerted action will it be possible to assure the production, reproduction, and use of knowledge that is crucial to the advancement of global health. PMID- 18395583 TI - Public-sector health-care reforms that work? A case study of the US Veterans Health Administration. PMID- 18395584 TI - Black lymph nodes--and a colourful skin. PMID- 18395585 TI - Special issue: redox regulation of protein folding. Preface. PMID- 18395586 TI - Witness to a revolution. PMID- 18395582 TI - Premenstrual syndrome. AB - Most women of reproductive age have some physical discomfort or dysphoria in the weeks before menstruation. Symptoms are often mild, but can be severe enough to substantially affect daily activities. About 5-8% of women thus suffer from severe premenstrual syndrome (PMS); most of these women also meet criteria for premenstrual dysphoric disorder (PMDD). Mood and behavioural symptoms, including irritability, tension, depressed mood, tearfulness, and mood swings, are the most distressing, but somatic complaints, such as breast tenderness and bloating, can also be problematic. We outline theories for the underlying causes of severe PMS, and describe two main methods of treating it: one targeting the hypothalamus pituitary-ovary axis, and the other targeting brain serotonergic synapses. Fluctuations in gonadal hormone levels trigger the symptoms, and thus interventions that abolish ovarian cyclicity, including long-acting analogues of gonadotropin-releasing hormone (GnRH) or oestradiol (administered as patches or implants), effectively reduce the symptoms, as can some oral contraceptives. The effectiveness of serotonin reuptake inhibitors, taken throughout the cycle or during luteal phases only, is also well established. PMID- 18395590 TI - Wow factor. PMID- 18395593 TI - 'Sniffing' for airways. PMID- 18395594 TI - Looking the part: why our appearance matters when we're responding to calls. PMID- 18395595 TI - Employees with 'INK': the need for a policy on tattoos & other body art. PMID- 18395597 TI - Write it right keeping your PCR clinical & factual. PMID- 18395598 TI - The hours: how many shifts are we really working? PMID- 18395599 TI - What current studies mean to EMS. PMID- 18395600 TI - From idea to innovation: tips for engineering safety, comfort & communications into your next ambulance. PMID- 18395601 TI - The secrets to ambulance financing: what to watch for & watch out for. PMID- 18395602 TI - The ins & outs of patient meds: knowing about common medications leads to improved patient care. PMID- 18395603 TI - Siren food. PMID- 18395605 TI - WITHDRAWN: Peripheral mGluR5 antagonist attenuated craniofacial muscle pain and inflammation but not mGluR1 antagonist in lightly anesthetized rats. AB - The present study investigated the role of peripheral group I metabotropic glutamate receptors (mGluRs) in MO-induced nociceptive behaviour and inflammation in the masseter muscles of lightly anesthetized rats. Experiments were carried out on male Sprague-Dawley rats weighing 300-400 g. After initial anesthesia with sodium pentobarbital (40 mg/kg, i.p.), one femoral vein was cannulated and connected to an infusion pump for intravenous infusion of sodium pentobarbital. The rate of infusion was adjusted to provide a constant level of anesthesia. Mustard oil (MO, 30 microl) was injected into the mid-region of the left masseter muscle via a 30-gauge needle over 10s. After 30 microl injection of 5, 10, 15, or 20% MO into the masseter muscle, the total number of hindpaw shaking behaviour and extravasated Evans' blue dye concentration in the masseter muscle were significantly higher in the MO-treated group in a dose-dependent manner compared with the vehicle (mineral oil)-treated group. Intramuscular pretreatment with 3 or 5% lidocaine reduced MO-induced hindpaw shaking behaviour and increases in extravasated Evans' blue dye concentration. Intramuscular pretreatment with 5mM MCPG, non-selective group I/II mGluR antagonist, or MPEP, a selective group I mGluR5 antagonist, produced a significant attenuation of MO-induced hindpaw shaking behaviour and increases in extravasated Evans' blue dye concentration in the masseter muscle while LY367385, a selective group I mGluR1 antagonist, did not affect MO-induced nociceptive behaviour and inflammation in the masseter muscle. These results indicate that peripheral mGluR5 plays important role in mediating MO-induced nociceptive behaviour and inflammation in the craniofacial muscle. PMID- 18395604 TI - Use of a modified prepulse inhibition paradigm to assess complex auditory discrimination in rodents. AB - Prepulse inhibition (PPI; also termed startle reduction or reflex modification, see Ref. [H.S. Hoffman, J.R. Ison, Reflex modification in the domain of startle: I. Some empirical findings and their implications for how the nervous system processes sensory input, Psychol. Rev. 87 (1980) 175-189]) provides an efficient and accurate method to assess both simple and complex acoustic discrimination in rodents [J.R. Ison, G.R. Hammond, Modification of the startle reflex in the rat by changes in the auditory and visual environments, J. Comp. Physiol. Psychol. 75 (1971) 435-452]. Assessment of acoustic processing using PPI is less time consuming than operant conditioning paradigms, allows for the testing of many subjects simultaneously, and largely eliminates confounds due to motivation and attention [M. Clark, G. Rosen, P. Tallal, R.H. Fitch, Impaired processing of complex auditory stimuli in rats with induced cerebrocortical microgyria, J. Cog. Neurosci. 12 (2000) 828-839]. Moreover, PPI procedures allow for data acquisition from the first day of testing, and can be used on rats as young as P14-15 [J.T. Friedman, A. Peiffer, M. Clark, A. Benasich, R.H. Fitch, Age and experience related improvements in gap detection in the rat, Dev. Brain Res. 152 (2004) 83 91; M. McClure, S. Threlkeld, G. Rosen, R.H. Fitch, Rapid auditory processing and learning deficits in rats with P1 versus P7 neonatal hypoxic-ischemic injury, Behav. Brain Res. 172 (2006) 114-121; S.W. Threlkeld, M.M. McClure, G.D. Rosen, R.H. Fitch, Developmental timeframes for the induction of microgyria and rapid auditory processing deficits in the rat, Brain Res. 1109 (2006) 22-31]. For these and additional reasons, the PPI paradigm has more recently been adapted to the assessment of complex acoustic discrimination (tone sequences and FM sweeps), and applied to the study of normally developing as well as neuropathologically affected rodent populations. The purpose of the current review is to provide a background on the PPI paradigm, and to summarize what has been learned more recently using modified versions of PPI with rodent models. PMID- 18395606 TI - Muscle synergies involved in shifts of the center of pressure while standing on a narrow support. AB - We investigated multi-muscle synergies during preparation to push a load forward and their changes with different support conditions. We hypothesized that the subjects show unchanged mode structure and would be able to form multi-mode COP stabilizing synergies while standing on an unstable board. Eight healthy subjects participated in the study. Standing subjects performed load-pushing tasks under two conditions, "normal support" and "ML narrow support". Electromyographic (EMG) signals of 12 postural muscles were recorded and analyzed. The participants also performed standard tasks associated with releasing a load. These trials were used to identify muscle groupings (M-modes) associated with shifts of the center of pressure (COP) and relations between small changes in the M-modes and COP shifts in different support conditions. The subjects showed unchanged mode structure across different support conditions. The framework of the uncontrolled manifold hypothesis was used to partition the EMG variance across load-pushing trials into two components that kept constant and changed the COP coordinates in the anterior posterior (AP) direction. This analysis has allowed us associate changes in the contribution of muscles with COP shifts under different support conditions. Different time profiles of the synergies were observed related to the COP shifts across conditions. This outcome supports a view that indices of multi-muscle (multi-M-mode) synergies can show anticipatory changes in preparation for a predictable perturbation. PMID- 18395607 TI - Architectonic subdivisions of the amygdalar complex of a primitive marsupial (Didelphis aurita). AB - The architecture of the amygdaloid complex of a marsupial, the opossum Didelphis aurita, was analyzed using classical stains like Nissl staining and myelin (Gallyas) staining, and enzyme histochemistry for acetylcholinesterase and NADPH diaphorase. Most of the subdivisions of the amygdaloid complex described in eutherian mammals were identified in the opossum brain. NADPH-diaphorase revealed reactivity in the neuropil of nearly all amygdaloid subdivisions with different intensities, allowing the identification of the medial and lateral subdivisions of the cortical posterior nucleus and the lateral subdivision of the lateral nucleus. The lateral, central, basolateral and basomedial nuclei exhibited acetylcholinesterase positivity, which provided a useful chemoarchitectural criterion for the identification of the anterior basolateral nucleus. Myelin stain allowed the identification of the medial subdivision of the lateral nucleus, and resulted in intense staining of the medial subdivisions of the central nucleus. The medial, posterior, and cortical nuclei, as well as the amygdalopiriform area did not exhibit positivity for myelin staining. On the basis of cyto- and chemoarchitectural criteria, the present study highlights that the opossum amygdaloid complex shares similarities with that of other species, thus supporting the idea that the organization of the amygdala is part of a basic plan conserved through mammalian evolution. PMID- 18395609 TI - Colocalization of dopamine and GABA in spinal cord neurones in the sea lamprey. AB - In this study, double immunofluorescence methods were used to investigate possible colocalization of the neurotransmitters dopamine [DA] and GABA in rostral spinal cord neurones in the upstream migrating adult sea lamprey (Petromyzon marinus). Double immunofluorescence revealed that all the DA immunoreactive (ir) cerebrospinal fluid-contacting (CSF-c) cells, approximately 30% of the medioventral DA-ir cells, and most of the DA-ir cells located in the grey lateral to the central canal were also GABA-ir. The results also revealed some DA-ir cells located dorsally to the central canal, which increases the number of dopaminergic cell types known in lamprey. Double-labelled fibres were mainly distributed in the ventral column, and double-labelled boutons contacted some dorsal GABA-ir CSF-c cells, as well as some non-CSF-c GABA-ir cells and ventromedial dendrites of motoneurones. The findings reveal colocalization of dopamine and GABA in some cells and fibres, which suggests co-release of these substances in some synaptic terminals. Although dopaminergic/GABAergic CSF-c cells have been reported in some other vertebrates, the other double-labelled spinal populations appear exclusive to lampreys. PMID- 18395608 TI - Modulation of vascular endothelial growth factor (VEGF) expression in motor neurons and its electrophysiological effects. AB - Previous studies have shown that VEGF expression in forebrain increases after experimental manipulations that increase neuronal activity. One question is whether this also occurs in motor neurons. If so, it could be potentially advantageous from a therapeutic perspective, because VEGF prevents motor neuron degeneration. Therefore, we asked whether endogenous VEGF expression in motor neurons could be modulated. We also asked how VEGF exposure would influence motor neurons using electrophysiology. Immunocytochemistry showed that motor neuron VEGF expression increased after a stimulus that increases neuronal and motor activity, i.e., convulsive seizures. The increase in VEGF immunoreactivity occurred in all motor neuron populations that were examined 24h later. This effect was unlikely to be due to seizure-induced toxicity, because silver degeneration stain did not show the typical appearance of a dying or dead neuron. To address the effects of VEGF on motor neuron function, VEGF was applied directly to motor neurons while recording intracellularly, using a brainstem slice preparation. Exposure to exogenous VEGF (200 ng/ml) in normal conditions depressed stimulus-evoked depolarization of hypoglossal motor neurons. There was no detectable effect of VEGF on membrane properties or firing behavior. We suggest that VEGF is upregulated in neurons when they are strongly activated, and VEGF depresses neuronal excitation as a compensatory mechanism. Failure of this mechanism may contribute to diseases that involve a dysregulation of VEGF, excessive excitation of motor neurons, and motor neuron loss, such as amyotrophic lateral sclerosis (ALS). PMID- 18395610 TI - TMS of the posterior parietal cortex delays the latency of unpredictable saccades but not when they are combined with predictable divergence. AB - This study tests the influence of transcranial magnetic stimulation (TMS) of the posterior parietal cortex (PPC) on the initiation of horizontal and vertical saccades, alone or combined with a predictable divergence. A gap paradigm was used; TMS was applied 100 ms after target onset. TMS of the left PPC increased the latency of unpredictable rightward saccades, while TMS of the right PPC increased the latency of unpredictable downward saccades. Yet, when unpredictable saccades were combined with predictable divergence, neither component was affected. We suggest that in the latter case, the initiation of both components was taken in charge by another area, e.g. frontal. Thus, even when one component was predictable, a common mechanism controls the initiation of both components. The results confirm that TMS only modifies the latency when the cortical area stimulated is involved in the triggering of the eye movement. PMID- 18395611 TI - Neuroprotective effect of aminoguanidine on iron-induced neurotoxicity. AB - Iron is a commonly used metal to induce neuronal hyperactivity and oxidative stress. Iron levels rise in the brain in some neurodegenerative disorders such as Parkinson's and Alzheimer's diseases. A body of evidence indicates a link between neuronal death and nitric oxide. The present study was performed to investigate whether nitric oxide produced by inducible nitric oxide synthase is involved in iron-induced neuron death. For this purpose rats were divided into four groups: control, iron, aminoguanidine and iron+aminoguanidine. Animals in iron and iron+aminoguanidine groups received intracerebroventricular FeCl3 injection (200 mM, 2.5 microl). Rats belonging to control and aminoguanidine groups received the same amount of saline into the cerebral ventricles. All animals were kept alive for 10 days following the operation and animals in aminoguanidine and iron+aminoguanidine groups received intraperitoneal aminoguanidine injections once a day (100mg/kg day) during this period. After 10 days, rats were perfused intracardially under deep urethane anesthesia. Removed brains were processed using the standard histological techniques. The total numbers of neurons in hippocampus of all rats were estimated with the unbiased stereological techniques. It was found that aminoguanidine decreased mean neuron loss from 43.4% to 20.3%. Results of the present study suggest that aminoguanidine may attenuate the neurotoxic effects of iron by inhibiting inducible nitric oxide synthase. PMID- 18395612 TI - Acute baroreceptor unloading evokes Fos expression in anesthetized rat brain. AB - Functional organization of brain pathways subserving the baroreceptor reflex was investigated by mapping immunoreactivity of Fos protein, a neuronal activity marker, in response to acute baroreceptor unloading in anesthetized rats. Compared with normal control and sham operation, sinoaortic denervation (SAD) evoked a distinctive pattern of Fos expression in the nucleus tractus solitarii (NTS), the primary termination of baroreceptor afferents. The sinoaortic denervation also induced a prominent and reproducible Fos expression in specific regions of the brainstem and forebrain, which receive direct or indirect inputs from the nucleus tractus solitarii. These brain regions included the rostral ventrolateral medulla (RVLM), lateral parabrachial nucleus (LPBN), hypothalamic paraventricular nucleus (PVN), supraoptic nucleus (SON), and central amygdaloid nucleus (CeA). These findings help us to identify brain regions that are specifically responsive to decreased arterial baroreceptor activity, without the accompanying confounding variables of behavioral arousal or stress. PMID- 18395613 TI - Increased thirst and drinking in Huntington's disease and the R6/2 mouse. AB - While Huntington's disease (HD) is a condition that primarily involves the basal ganglia, there is evidence to suggest that the hypothalamus is also affected. Because the osmoreceptors regulating thirst are situated in the circumventricular region of the hypothalamus, we were interested in whether altered thirst is a part of the HD phenotype. We used the LABORAS behavioural monitoring system and water consumption to show that drinking behaviour was abnormal in R6/2 mice. By 10 weeks of age, R6/2 mice spent significantly more time drinking and drank a greater volume than their wild-type (WT) littermates. The numbers of immunoreactive vasopressin neurons in the paraventricular nucleus (PVN) of the hypothalamus in R6/2 mice were significantly decreased from 8 weeks of age, suggesting that the change in drinking behaviour may be the result of hypothalamic dysfunction. We gave a xerostomia (dry mouth) questionnaire to HD patients and control subjects, and also measured their urine osmolality and serum vasopressin. The mean total xerostomia score was significantly higher in HD patients than in controls, indicating greater thirst in HD patients. Urine osmolality was unaffected in HD patients up to clinical stage III, and none of the patients had diabetes. However, serum vasopressin was increased, suggesting a dysregulation in the control of hypothalamic vasopressin release. A dry mouth can affect taste, mastication and swallowing, all of which may contribute to the significant weight loss seen in both HD patients and R6/2 mice, as can dehydration. We suggest that increased thirst may be an important and clinically relevant biomarker for the study of disease progression in HD. PMID- 18395614 TI - CSF phosporylated TAU protein levels correlate with cerebral glucose metabolism assessed with PET in Alzheimer's disease. AB - One major goal of drug development would be the establishment of biomarkers as objective indicators of normal biological and pathogenetic processes, or pharmacological response to a therapeutic intervention. A potential approach is to investigate proteins in CSF linked to key neuropathological features of Alzheimer's disease (AD). Recently CSF phosphorylated-Tau (p-Tau) levels have been reported to reflect neurofibrillary changes within the brain of patients with AD, however the use of serial CSF investigations in order to monitor the disease progression is not applicable. PET with FDG reveals characteristic patterns in AD patients, however so far no correlation between in vivo metabolic information and pathological features has been reported. In the present study, we tested whether CSF Tau levels correlate with metabolic rate for glucose consumption in a cohort of 28 AD patients. We found a statistically significative correlation between both CSF total and p-TAU protein and relative metabolic indexes obtained from 18FDG-PET scans in parietal, temporal and occipital lobes bilaterally. These results indicate the existence of a correlation between impairment of cerebral metabolism, estimated throughout FDG-PET, and CSF Tau protein levels. PMID- 18395615 TI - Wakefulness-sleep transition: emerging electroencephalographic similarities with the rapid eye movement phase. AB - The covert-rapid eye movement (REM) sleep hypothesis of dreaming suggests that elements of REM sleep emerge during sleep onset, leading to vivid hypnagogic imagery. We tested the physiological part of this hypothesis by analysing scalp recorded electroencephalograms of 15 human subjects during wake-sleep transition and subsequent night time sleep. Wake-sleep transition was categorised semi automatically as alpha activity, alpha dropout and as early Stage 2 sleep. The slow oscillation, the slow and the fast subdivisions of the delta and the theta frequencies respectively, as well as alpha and sigma bands were analysed. The similarity of individual-specific wake-sleep transition periods and the whole night Stage 2 or REM sleep periods was expressed in a composite similarity measure covering the spectral power of all analysed frequency bands and in frequency-specific similarities related to power values in single bands. A significant increase in composite similarity with the whole night REM sleep emerged in the period of alpha dropout and diminished in early Stage 2 sleep. The alpha dropout period was more similar to whole night REM sleep than to whole night Stage 2 sleep. These region-independent effects were mirrored in region specific manner by frequency bands of the delta-slow theta range. Findings are in accordance with the covert REM sleep hypothesis, with previous electrocorticographic results and with the frequency range of the sawtooth waves in humans. PMID- 18395616 TI - Mu-calpain mediates hippocampal neuron death in rats after lithium-pilocarpine induced status epilepticus. AB - Status epilepticus (SE) is a severe clinical manifestation of epilepsy which causes brain damage. The pathological process and underlying mechanisms involved in the programmed cell death (PCD) are still not fully clear. In the current study, rats were induced SE by lithium-pilocarpine administration. Our data showed hippocampal neurons death appeared at 6h after SE and sustained for 7 days. By blotting the activation of mu-calpain and its specific cleavage of nonerythroid alpha-spectrin (alphaSpII) (145 kDa) was evident at 1 and 3 days after SE, which coincided with Bid activation, apoptosis inducing factor (AIF) translocation and cytochrome c release from mitochondria, whereas, activated caspase-3 and caspase-3-specific fragments of alphaSpII (120 kDa) predominantly appeared at 5 and 7 days after SE. Moreover, MDL-28170, a calpain inhibitor, partially rescued the neuron death and attenuated the expression of activated mu calpain, cleavage of Bid (15 kDa), AIF translocation and cytochrome c release. Taken together, our study indicated that mu-calpain mediated hippocampal neuron PCD is prior to caspase-3 activation. It functioned via translocation of Bid, AIF and cytochrome c release. PMID- 18395617 TI - Induction of orientation-specific LTP-like changes in human visual evoked potentials by rapid sensory stimulation. AB - Recent research suggests that rapid visual stimulation can induce long-term potentiation-like effects non-invasively in humans. However, to date, this research has provided only limited evidence for input-specificity, a fundamental property of cellular long-term potentiation. In the present study we extend the evidence for input-specificity by investigating the effect of stimulus orientation. We use sine wave gratings of two different orientations to show that rapid visual stimulation can induce orientation-specific potentiation, as indexed by changes in the amplitude of a late phase of the N1 complex of the visual evoked potential. This result suggests that discrete populations of orientation tuned neurons can be selectively potentiated by rapid visual stimulation. Furthermore, our results support earlier studies that have suggested that the locus of potentiation induced by rapid visual stimulation is visual cortex. PMID- 18395618 TI - Neuroprotective effects of sodium orthovanadate after hypoxic-ischemic brain injury in neonatal rats. AB - Sodium orthovanadate (SOV), a competitive inhibitor of protein tyrosine phosphatases, is neuroprotective in adult animals following an ischemic event. The present study evaluated whether SOV might be protective in a rat pup hypoxic ischemic (HI) model. Seven-day-old rat pups had the right carotid artery permanently ligated followed by 140 min of hypoxia (8% oxygen). SOV 1.15, 2.3, 4.6, 9.2 or 18.4 mg/kg and vehicle were administered by i.p. injection at 5 min after reoxygenation. Brain damage was evaluated by weight loss of the right hemisphere at 22 days after hypoxia and by gross and microscopic morphology. SOV lowered blood glucose at doses of 1.15, 2.3 and 4.6 mg/kg and induced toxic effects at 9.2mg/kg. The doses of 2.3 and 4.6 mg/kg of SOV significantly reduced brain weight loss (p<0.05), but treatment with 1.15 or 9.2mg/kg did not. SOV 4.6 mg/kg also improved the histopathologic score and diminished the HI induced reduction of Akt and ERK-1/2 phosphorylation in the cortex (p<0.05) and increased the density of BrdU-positive cells in the subventricular zone (p<0.01). In conclusion, SOV has neuroprotective effects in the neonatal rat HI model partially mediated by activating Akt and ERK-1/2 pathways. PMID- 18395619 TI - Ascorbic acid supplementation could affect passive avoidance learning and memory in rat. AB - Ascorbic acid (vitamin C) is required for health and, in particular, its supplementation has beneficial effects in some pathological conditions. There are conflicting reports regarding the usefulness of ascorbic acid in the treatment of dementia. In this study, we investigated the effects of acute, short- and long term pre-training administration of ascorbic acid (60,120 mg/kg) on passive avoidance learning (PAL) and memory in rats. Retention test was done 24h after training. The results showed that acute injection of ascorbic acid had no significant effect on PAL. On the other hand, both in the short- and long-term ascorbic acid treated groups trials to acquisition were less than control group. Also, ascorbic acid prolonged the step-through latency (STL) and decreased the time spent in the dark compartment in retention test. Thus, it can be concluded that short- and long-term supplementation with ascorbic acid has facilitatory effects on acquisition and retrieval processes of passive avoidance learning and memory in rats. PMID- 18395620 TI - An organotellurium compound with antioxidant activity against excitotoxic agents without neurotoxic effects in brain of rats. AB - The glutamatergic system is an important target in many neurodegenerative diseases and for several neurotoxic drugs. Organotellurium compounds are often very good free radical scavengers' agents. Recently, we reported that diethyl-2 phenyl-2-tellurophenyl vinylphosphonate is a compound with low toxicity in vitro and in vivo, as well as also possesses antioxidant activity against iron-induced lipid peroxidation. The aim of this study was to evaluate in vitro the antioxidant and mitochondrial protective effect of this organotellurium compound against quinolinic acid (QA) and sodium nitroprusside (SNP), and to evaluate the in vitro actions of this organotellurium compound in the glutamatergic system in brain of rats. We observed that the telluro vinylphosphonate possess an antioxidant activity against QA and SNP at micromolar concentrations. When tested at antioxidant concentrations (from 2 to 10muM), the compound does not affect the mitochondrial viability and [3H]glutamate uptake in slices from cerebral cortex, hippocampus and striatum, [3H]glutamate release from synaptosomal preparations and [3H]glutamate binding in membrane preparation. Our data suggest that the telluro vinylphosphonate act as an antioxidant in the central nervous system in vitro with no effects on the glutamatergic system; nevertheless more studies in different models of brain injury must be performed in order to corroborate our findings. PMID- 18395621 TI - TrkA pathway(s) is involved in regulation of TRPM7 expression in hippocampal neurons subjected to ischemic-reperfusion and oxygen-glucose deprivation. AB - The transient receptor potential melastatin 7 (TRPM7) is recently revealed playing a key role in anoxic neuronal death. Meanwhile, nerve growth factor (NGF), through activating TrkA pathway, has been widely accepted as a crucial factor for neuronal survival during cerebral ischemia. In this study, middle cerebral artery occlusion (MCAO) for 1h and reperfusion for 5, 10, 20 and 30 h revealed an increasing up-regulation of TRPM7 expression in ipsilateral hippocampus after reperfusion and such a change reached a peak at 20 h, with high expression being maintained up to 30 h. Intracerebroventricular injection of NGF (500 ng) 30 min before MCAO, the expression of TRPM7 at 20 h after reperfusion was significantly reduced, and the effect of NGF was completely abolished by co injection of TrkA inhibitor K252a. Furthermore, TRPM7 was found residing on hippocampal neuronal body and process, and TrkA was concurrently observed on the cell body by immunofluorescence staining. In agreement with in vivo, pre incubation of cultured hippocampal neurons with NGF markedly down-regulated TRPM7 high expression at 20 h after oxygen-glucose deprivation (OGD), and this effect could be abrogated by K252a. In addition, when inhibitors wortmannin, U73122 and U0126 were introduced to block phosphatidylinositol-3 kinase (PI-3K), phospholipase C-gamma (PLC-gamma) and ras/mitogen-activated protein kinase (MAPK) pathways, respectively, only PI-3K inhibitor wortmannin substantially abolished NGF effects. These results suggest that TrkA, after being activated by NGF, can prevent up-regulation of TRPM7 expression in hippocampal neurons during reperfusion through PI-3K signal pathway. These findings open a new way for further investigations of the potential roles of TRPM7 and NGF in cerebral ischemia-reperfusion. PMID- 18395622 TI - Methylphenidate sensitization is prevented by prefrontal cortex lesion. AB - Methylphenidate (MPD), also known as Ritalin, is a widely used treatment for Attention Deficit Hyperactivity Disorder. Repeated administration of MPD causes dose-dependent sensitization. MPD binds to dopamine (DA) transporters, and DA, therefore, remain in the synaptic cleft for longer time, resulting in an indirect DA agonist effect. MPD affects neurotransmission in brain regions including the prefrontal cortex (PFC). The mechanisms of sensitization to MPD are not clear. The aim of this study was to investigate the role of prefrontal cortex in effects of acute and chronic MPD administration, using the open field assay and male Sprague-Dawley rats with bilateral electrolytic lesions of PFC. After 1 day of control recording, following saline injection, the animals were divided randomly into three groups, (1) an intact control group, (2) a sham group, and (3) a lesion group. Then, groups 2 and 3 underwent surgery, followed by 5 days of recovery. Recordings were resumed following 1 day of saline injection and following six consecutive daily injections of 2.5mg/kg MPD, 3 days of washout period, and another day of re-challenge injection of 2.5mg/kg MPD. Acute MPD elicited increases in locomotor activity, similar to those observed from intact animals, in both sham and lesion groups. The sham group was behaviorally sensitized while the PFC lesion group failed to exhibit behavioral sensitization. These results suggest that the PFC does not interfere with the acute effects of MPD on locomotor activity but is required for development of behavioral sensitization to MPD. PMID- 18395623 TI - Chronic disruption of circadian rhythms impairs hippocampal memory in the rat. AB - Circadian related disorders and alterations in sleep-wake patterns are common complaints in the elderly, especially those diagnosed with Alzheimer's disease. The negative physical and psychological effects resulting from chronic circadian disruption are numerous and appear to be positively correlated with the length of time an individual has suffered from a circadian disorder. In the current paper, we explore the effects of acute and chronic disruption of circadian rhythms on memory using a phase shifting schedule that can continually challenge the rats' circadian system by using repeated phase shifts and recovery sessions. We demonstrate a significant learning and memory deficit on a spatial version of the water maze task in the chronically phase shifted, but not in the acutely phase shifted animals. Moreover, we find no impairment in fear conditioning suggesting that chronic phase shifting predominantly affects hippocampal memory. We propose that chronic circadian disruption may play a role in the development of age related cognitive deficits and dementia in the elderly. PMID- 18395624 TI - Adolescent nicotine treatment changes the response of acetylcholine systems to subsequent nicotine administration in adulthood. AB - Nicotine alters the developmental trajectory of acetylcholine (ACh) systems in the immature brain, with vulnerability extending from fetal stages through adolescence. We administered nicotine to adolescent rats (postnatal days PN30-47) and then examined the subsequent response to nicotine given in adulthood (PN90 107), simulating plasma levels in smokers, and performing evaluations during nicotine treatment (PN105) and withdrawal (PN110, PN120 and PN130), as well as assessing persistent changes at 6 months of age (PN180). We measured nicotinic acetylcholine receptor (nAChR) binding, choline acetyltransferase (ChAT) activity, a marker for ACh terminals, and hemicholinium-3 (HC3) binding to the choline transporter, an index of ACh presynaptic activity. By itself, adolescent nicotine exposure evoked sex-selective deficits in cerebrocortical HC3 binding while elevating ChAT in young adulthood in striatum and midbrain. Nicotine given in adulthood produced profound nAChR upregulation lasting 2 weeks after discontinuing treatment, and decrements in cerebrocortical and striatal HC3 binding emerged during withdrawal, indicative of reduced ACh synaptic activity. For all three parameters, adolescent nicotine altered the responses to nicotine given in adulthood, producing both sensitization and desensitization that depended on sex and brain region, effects that parallel the disparate behavioral outcomes reported for these treatments. The interaction seen here for the impact of adolescent nicotine exposure on adult nicotine responses was substantially greater than that found previously for the effects of prenatal nicotine exposure on adult responses. Our findings thus reinforce the importance of adolescence as a critical period in which the future responsiveness to nicotine is programmed. PMID- 18395625 TI - Minimal-incision cardiac surgery. Introduction. PMID- 18395626 TI - Minimally invasive coronary artery bypass grafting. AB - Limited access, off-pump coronary artery bypass grafting for revascularization of all the various coronary arteries is an acceptable alternative to standard on pump coronary bypass grafting through sternotomy. A variety of small, targeted incisions are used to approach various coronary locations. Technical advances in conduit harvesting, stabilization, cardiac positioning devices, and anastomotic connectors have made these procedures more standardized and replicable. This has resulted in reduced morbidity as a consequence of less invasive approaches. These efforts have paved the way for the ultimate goal of same day surgical coronary revascularization. PMID- 18395627 TI - Minimally invasive valve surgery. AB - As alternatives to standard sternotomy, surgeons have developed innovative, minimally invasive approaches to conducting valve surgery. Through very small skin incisions and partial upper sternal division for aortic valve surgery and right minithoracotomy for mitral surgery, surgeons have become adept at performing complex valve procedures. Beyond cosmetic appeal, apparent benefits range from decreased pain and bleeding to improved respiratory function and recovery time. The large retrospective studies and few small prospective randomized studies are herein briefly summarized. The focus is then directed toward describing specific intraoperative technical details in current clinical use, covering anesthetic preparation, incision, mediastinal access, cardiovascular cannulation, valve exposure, and valve reconstruction. Finally, unique situations such as pulmonic valve surgery, reoperations, beating heart surgery, and robotics are discussed. PMID- 18395628 TI - Minimally invasive surgery with a partial sternotomy "J" approach. AB - The minimally invasive partial sternotomy "J" incision can be used for most isolated mitral valves, tricuspid valves, aortic valves, atrial septal defects, maze procedures, aortic repairs, and aortic valve reoperations. This article reviews the technical approaches and outcomes for various procedures. For 2,004 mitral valve repairs, our 30-day in-hospital mortality was 0.2% and for 1,103 aortic valve procedures, it was 0.8%. In addition to both a better cosmetic result and earlier return to work, the benefits include less blood loss, less pain, better respiratory function, and better 1-year survival for mitral valve procedures. Similar results have been obtained for aortic valve procedures. Thus, all patients undergoing isolated aortic or mitral valve repairs are offered a minimally invasive operation. PMID- 18395629 TI - Transcatheter percutaneous and transapical aortic valve replacement. AB - Transcatheter aortic valve implantation has evolved rapidly since the first successful procedure over 5 years ago. The femoral arterial and the left ventricular apical approaches are currently favored. Initial skepticism has to some degree given way to acceptance that the procedure might be a reasonable alternative for selected patients who might be at high risk with conventional thoracotomy and cardiopulmonary bypass. Uncertainty remains about outcomes, implications, durability, and the appropriate role for this new therapy. It appears likely that transcatheter valve implantation will become a more widely available and accepted therapeutic option. PMID- 18395630 TI - Minimally invasive surgical treatment of atrial fibrillation. AB - Surgical ablation for atrial fibrillation (AF) can now be successfully performed utilizing minimally invasive techniques, as surgeon experience with these approaches has increased, and technologic advances in minimally invasive instrumentation have been made. Surgical management of atrial fibrillation, as well as our minimally invasive approach, will be discussed in depth. PMID- 18395631 TI - Minimal incision congenital cardiac surgery. AB - Minimally invasive techniques have had limited application in congenital cardiac surgery, primarily due to the complexity of the defects, small working area, and the fact that most defects require exposure to intracardiac structures. Advances in cannula design and instrumentation have allowed application of minimal incision techniques but in most cases, cardiopulmonary bypass is still required. Image guided surgery, which uses noninvasive imaging to guide intracardiac procedures, holds the promise of permitting performance of reconstructive surgery in the beating heart in children. PMID- 18395632 TI - 3D echocardiographic visualization for intracardiac beating heart surgery and intervention. AB - Three-dimensional echocardiography has emerged as an essential tool for visualizing cardiac anatomy and for making more accurate measurements of cardiac structure and function. Recently, improvements in 3D beam-forming and transducer technologies have allowed higher resolution imaging from a transesophageal echocardiographic probe. This is creating new avenues for real-time visualization of intracardiac procedures without the need for cardiopulmonary bypass or opening the beating heart. Evolutions in visualization will allow a wider array of reparative procedures to be performed minimally invasively within a beating heart. PMID- 18395633 TI - Real-time magnetic resonance imaging guidance for cardiovascular procedures. AB - Magnetic resonance imaging (MRI) of the cardiovascular system has proven to be an invaluable diagnostic tool. Given the ability to allow for real-time imaging, MRI guidance of intraoperative procedures can provide superb visualization, which can facilitate a variety of interventions and minimize the trauma of the operations as well. In addition to the anatomic detail, MRI can provide intraoperative assessment of organ and device function. Instruments and devices can be marked to enhance visualization and tracking, all of which is an advance over standard X ray or ultrasonic imaging. PMID- 18395634 TI - Real-time percutaneous optical imaging of anatomical structures in the heart through blood using a catheter-based infrared imaging system. AB - The ability to optically image structures and instrumentation within the heart during procedures is limited by the presence of blood in the field. The goal of our research was to design, develop, and evaluate technology for a catheter-based optical imaging system that enables intracardiac and intravascular visualization in real time through blood. Based on Mie optical scattering theory, imaging through blood using infrared light was theoretically feasible, but scattering in the near-infrared wavelengths (1100 to 1300 nm) generated substantial noise in the image despite relatively low absorption. Using illumination between 1550 and 1650 nm provided better images, as the effect of scattering is less while the effect of absorption is greater. Absorption losses can be overcome by increasing light intensity. Infrared (IR) transmitting endoscopes were constructed using novel flexible illumination and imaging bundles. Endoscope designs, all 7.5 Fr. in outer diameter, were used to obtain images of the coronary sinus, tricuspid valve, and great vessels in 25 pigs, 16 dogs, 1 calf, and 1 sheep. Imaging was successful in all 43 animals, but the coronary sinus was not always visualizable. After obtaining FDA 510(k) approval, the device was used to acquire images in 50 patients during placement of electrophysiologic leads via right heart catheterization. Clinical experience demonstrates successful visualization in the heart in 45 patients, although coronary sinus images were obtained only in 39 patients. High heart rates, large dilated hearts, and problems with catheter design prevented visualization in all patients. On occasion, it was possible to visualize the tricuspid valve. Infrared endoscopy allows for visualization of intimal surfaces of blood vessels, cardiac chambers, and valves through flowing blood. While technical challenges remain, the feasibility of the approach has been demonstrated. PMID- 18395635 TI - Management of complications of lung resection. Introduction. PMID- 18395636 TI - Complications in patients with severe emphysema. AB - Thoracic surgeons frequently evaluate patients with severe emphysema and concomitant pathology requiring pulmonary resection. There are no absolute guidelines defining the suitability of a given patient for resection. In this review, we outline our approach to evaluating and treating patients with severe emphysema in need of resection. We describe the lessons learned from lung volume reduction surgery and apply that knowledge to the care of the patient with severe emphysema. Careful preoperative evaluation of the patient's lung anatomy, distribution of emphysematous changes in the lung, and overall health is essential to identifying the appropriate candidate for resection and avoiding postoperative complications. PMID- 18395637 TI - Complications of thoracoscopic pulmonary resection. AB - Thoracoscopic strategies are becoming increasingly utilized in the management of patients with thoracic disease processes, including primary pulmonary malignancy, secondary pulmonary malignancy, granulomatous lung disease, and pleural processes. Although minimally invasive approaches have been demonstrated to improve outcomes and reduce complications, as compared to the conventional approach, the prevention, early recognition, and effective management of complications after thoracoscopic pulmonary resection are still critical factors in optimizing outcomes. PMID- 18395638 TI - Complications of extrapleural pneumonectomy. AB - Extrapleural pneumonectomy (EPP) is a technically demanding operation that includes two body cavities within the operative field. Avoidance of postoperative deaths requires an experienced team that can identify common morbidities. The ability to recognize and intervene correctly in the early phases of postoperative complications can avoid deaths. Although our group has experienced only 20 deaths following 496 EPPs (4% mortality), there were 328 patients (66%) who suffered morbidity. We have drawn upon our single-institution experience to describe the most common source of morbidity following EPP and to offer successful interventions. PMID- 18395639 TI - Anatomical lung resection after neoadjuvant chemoradiotherapy. AB - The critical role for anatomical lung resection -- segmentectomy, lobectomy, pneumonectomy -- in the treatment of Stage I and II non-small cell lung cancer is undisputed. In contrast, the primacy of surgery in the management of Stage III disease is not established. Increasingly, however, the multimodality approach to locally advanced lung cancer has gained acceptance, and the integration of surgery into the treatment algorithms for Stage III cancers, particularly N2 spread, has evolved. Herein, the important steps in this evolution are defined. The concept of induction or neoadjuvant chemoradiotherapy followed by resection is emphasized, and evidence supporting surgery's therapeutic value in this schema is provided. Our center's strategy for the successful and safe delivery of trimodality care is comprehensively outlined. PMID- 18395640 TI - Bronchial stump dehiscence: update on prevention and management. AB - Bronchial stump dehiscence is a serious complication and is associated with important morbidity and increased mortality. Because of this, efforts to prevent and treat these occurrences are essential. Etiology, risk factors, diagnosis and prevention, as well as current treatment options with an emphasis on surgical repair with muscle flaps are reviewed and discussed. PMID- 18395641 TI - Respiratory failure following pulmonary resection. AB - Improvements in the perioperative management of the patient undergoing pulmonary resections have reduced postoperative complication rates steadily in the last several decades. However, postresection respiratory failure, particularly lung injury with no discernible cause, remains a major cause of morbidity and mortality. Because the incidence of this entity is relatively low, the terminology, pathogenesis, and optimal management are poorly delineated in the literature. The purpose of this review is to describe the criteria used to define postresection lung injury, discuss the possible etiologic factors, and outline currently available treatment strategies. PMID- 18395644 TI - Exercise for preventing childhood obesity. AB - Childhood obesity is a key public health issue in the United States and around the globe in developed and developing countries. Obese children are at increased risk of acute medical illnesses and chronic diseases-in particular, osteoarthritis, diabetes mellitus, and cardiovascular disease, which can lead to poor quality of life; increased personal and financial burden to individuals, families, and society; and shortened lifespan. Physical inactivity and sedentary lifestyle are associated with being overweight in children and adults. Thus it is imperative to consider exercise and physical activity as a means to prevent and combat the childhood obesity epidemic. Familiarity with definitions of weight status in children and health outcomes like metabolic syndrome is crucial in understanding the literature on childhood obesity. Exercise and physical activity play a role in weight from the prenatal through adolescent time frame. A child's family and community impact access to adequate physical activity, and further study of these upstream issues is warranted. Recommended levels of physical activity for childhood obesity prevention are being developed. PMID- 18395645 TI - Role of the adolescent preparticipation physical examination. AB - With the exception of Rhode Island, all states require high school athletes to undergo a preparticipation examination. These examinations may vary from state to state, however. This article covers the basics of the history, physical examination, special tests, and issues surrounding clearance for various diagnoses. PMID- 18395646 TI - Strength training recommendations for the young athlete. AB - In recent years there has been a significant increase in the number of youth participating in organized and competitive sports. Recent studies have supported the participation of preadolescent athletes in strength training to improve health and performance in sports. This article presents the most recent data available to help youth develop a safe and effective strength training program. Variables, such as the recommended rate of progression, the number of sets and repetitions an athlete should perform on each exercise, and how often an athlete needs to workout to avoid loss of strength achieved during a period of strength training are presented. PMID- 18395647 TI - Adolescent sports concussion. AB - Approximately 2 million sports and recreation concussive injuries occur per year in the United States, which may be an underestimate because of inconsistent data reporting. The field of concussion management has evolved rapidly over the last 10 years, and with these advances comes new understanding of the significant symptomatic and cognitive impairments of concussion. These sequelae are more fully realized and may last longer than previously thought. Data have emerged regarding pathophysiology of concussion, risk factors, outcome, effects of repetitive injury, subtypes of concussive injury, and treatment protocols. This evidence calls for more conservative management of concussion, particularly in younger athletes, and demonstrates the shortcomings of concussion guidelines. PMID- 18395648 TI - Shoulder and elbow injuries in the adolescent athlete. AB - The shoulder and elbow represent two of the most commonly injured joints in the adolescent population. Specific injuries vary by sport and can involve various structures, depending on the mechanism of injury. Unlike the adult shoulder, the immature skeletal structure of the adolescent athlete can lead to several unique injuries. By understanding the special demands placed on the immature shoulder, the sports physician can more effectively treat the resultant injury. This article reviews the diagnosis and management of unique injuries to the shoulder and elbow in the adolescent athlete. PMID- 18395649 TI - Low back pain in the adolescent athlete. AB - Low back pain is a common problem among young athletes. These individuals are at risk for significant structural injuries or nonmechanical problems that can be associated with their symptoms. Any athlete who has severe, persisting, or activity-limiting symptoms must be evaluated thoroughly. Clinicians must have a working knowledge of the developmental issues, injury patterns, and particular conditions that may affect a given athlete and be able to work with patients in addition to families, coaches, trainers, and others involved in the care and training of the injured athlete. PMID- 18395650 TI - Examination and treatment of pediatric injuries of the hip and pelvis. AB - Injuries to the hip and pelvis are the least common of lower extremity injuries in youth sports, but include many of the more serious conditions. This article describes the bone and soft-tissue conditions of the hip and pelvis that may present to health care providers caring for the pediatric and adolescent sporting population. The article discusses epidemiology, mechanisms, clinical presentation, evaluation, and treatment options. PMID- 18395651 TI - Acute knee injuries in skeletally immature athletes. AB - The knee is the body part most commonly injured as a consequence of collisions, falls, and overuse occurring from childhood sports. The number of sports-related injuries is increasing because of active participation of children in competitive sports. Children differ from adults in many areas, such as increased rate and ability of healing, higher strength of ligaments compared with growth plates, and continued growth. Growth around the knee can be affected if the growth plates are involved in injuries. This article discusses fractures, anterior and posterior cruciate ligament injuries, and meniscal and patellar conditions. PMID- 18395652 TI - Common injuries of the foot and ankle in the child and adolescent athlete. AB - A myriad problems in the foot and ankle are specific to child and adolescent athletes. The anatomy of young athletes with respect to the presence of a growth plate makes their injury patterns different from those seen in adults. The main general injury patterns seen in the feet and ankles of children are related to growth and development or occur from overuse syndromes or acute trauma. In this article we outline in an anatomically oriented manner most of the common problems in this population. PMID- 18395653 TI - Nutritional requirements of the child and teenage athlete. AB - There has been an explosion in sports participation, especially for women, in the last 35 years mainly because of Title IX. In 2005-2006, nearly 3 million girls and 4.2 million boys participated in high school athletics, and many more participated in club sports and recreational activities. On the other end of the spectrum, the prevalence of obesity in the United States is at an all-time high. Proper nutrition in combination with the appropriate amount of physical activity is of paramount importance for this era of adolescents. PMID- 18395654 TI - Psychologic stress related to injury and impact on sport performance. AB - Injury rates are high among children and adolescent athletes. Psychosocial stressors, such as personality, history of stressors, and life event stress can influence injury occurrence. After injury, those same factors plus athletic identity, self-esteem, and significant others-such as parents, coaches, and teammates-can affect injury response, recovery and subsequent sport performance. Goal setting, positive self-talk, attribution theory, and relaxation or mental imagery are psychologic interventions that can help injured athletes cope with psychosocial stressors. Medical professionals should be aware of the potential influence that psychosocial stressors and psychologic interventions can have on injury occurrence, injury recovery, and sport performance. PMID- 18395655 TI - Long term outcomes in pediatric surgery. Preface. PMID- 18395656 TI - Contemporary outcomes research: tools of the trade. AB - Outcomes are, simply put, why a surgeon comes to work each day. For decades, surgeons have insisted on a regular self-examination of outcomes to ensure the optimal treatment of our patients. Clinical research in pediatric surgery has largely subsisted on outcome analysis as it relates to the rudimentary end-result of an operation, utilizing variables such as mortality, operative time, specific complication rates, and hospital length of stay to name a few. Recently, outcomes research has become a more complex endeavor. This issue of Seminars in Pediatric Surgery addresses a wide array of these newfound complexities in contemporary outcomes research. The purpose of this review is to assist the pediatric surgeon in understanding the tools that are used in contemporary outcomes research and to be able to use this information to ask new questions of our patients and ourselves as we continue to strive for excellence in caring for sick infants and children. PMID- 18395657 TI - Imperforate anus: long- and short-term outcome. AB - Anorectal malformations are common anomalies observed in neonates. Survival of these babies is currently achieved in most cases and improvements in operative technique, patient care, and better follow-up have led to improved functional results. A new, simplified classification system (Krickenbeck classification) and method of functional assessment has led to an improved understanding of these anomalies and has allowed for a better comparison of outcomes. Following successful anatomical repair and appropriate programs of bowel care, socially acceptable continence can be achieved in a majority of patients, especially those with an intact sacrum. PMID- 18395658 TI - The Congenital Diaphragmatic Hernia Study Group: a voluntary international registry. AB - The Congenital Diaphragmatic Hernia Registry (CDHR) was established to collect data on all infants treated at participating institutions with congenital diaphragmatic hernia (CDH) to assess therapy and improve outcome. Since 1995, the CDHR has accumulated data on over 4000 infants. The collective efforts of the CDH Study Group have provided valuable information regarding the efficacy of various therapeutic interventions and have tried to establish predictors of outcome. Because CDH is an uncommon, heterogeneous structural anomaly with a wide spectrum of severity, individual institutions may vary significantly in their experiences and treatment approaches. International multicenter registries, like the CDHR, are useful to provide clinically relevant direction by accumulating far more data on a large patient population than any single institution might. PMID- 18395659 TI - Neonatal necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) remains a major cause of morbidity and death in neonates. The 30% to 50% mortality rate for NEC with perforation has not changed appreciably in the past 30 years. The critical relevant outcomes following NEC include survival, gastrointestinal function, and neurodevelopmental status. In each of these areas, initial anecdotal and case-series analysis has been followed by studies using more sophisticated methods of analysis. The single most important predictor of outcome, besides gestational age, is whether or not the disease has progressed to the point requiring surgical intervention. Patients with NEC requiring operation have a high mortality. Moreover, the vast majority of morbidity following NEC occurs in the patients who survive following operation. The purpose of this review is to examine the evolution of evidence regarding outcomes for patients with NEC and to provide an update on our current state of knowledge. PMID- 18395660 TI - Outcomes in pediatric trauma care. AB - Variation in clinical practice patterns has attracted the attention of specialty organizations, payers, government health agencies, and the public. Such variation raises concerns about efficacy and cost relative to the care provided. Consequently, the establishment of national benchmarks has become an increasing priority in trauma care as well as elsewhere in the health sector. Comparing treatment of pediatric injury by hospital type or physician expertise has often created more controversy than conformity. Three key components that help define quality of care include infrastructure, process, and outcome. This report will highlight outcomes studies in pediatric trauma care, often comparing outcomes by physician and hospital expertise. We will discuss pediatric trauma mortality and outcomes in abdominal, closed head, and orthopedic injuries with an emphasis on functional outcomes. Much of the data are derived from large regional and national databases, which are increasingly available and useful in the analysis of specific aspects of our health care delivery system. PMID- 18395661 TI - Biliary atresia: service delivery and outcomes. AB - Biliary atresia is a complex disorder dependent on multidisciplinary management. A series of comprehensive national audits in the United Kingdom and France exposed a clear relationship between center volume and clinical outcomes. Different models were adopted in each country in an attempt to improve results. In the United Kingdom, the management of biliary atresia was centralized to three specialist units in 1999, whereas in France, a strategy of decentralized management with closer inter-unit cooperation was adopted in 1997. Both policy changes led to improved outcomes for infants with biliary atresia, but only centralization improved the overall results of Kasai portoenterostomy. Other countries have adopted alternative systems of audit based on voluntary registries, but the impact of these on clinical outcomes at a national level remains unknown. The utility of monitoring tools in assessing performance in biliary atresia, the importance of risk stratification, and the need for standardized definitions of outcome are highlighted. PMID- 18395662 TI - Outcomes following liver transplantation. AB - As the field of Liver Transplantation has matured, survival alone is no longer an acceptable single metric of success. This chapter explores the impact of the PELD system for donor organ allocation, surgical modification of donor organs, living donation, and long-term transplant-related complications on overall quality of life and outcome. Strategies to improve survival, overall outcome, and health related quality of life in long-term recipients are outlined. PMID- 18395663 TI - Developing a NSQIP module to measure outcomes in children's surgical care: opportunity and challenge. AB - Under the guidance of the American College of Surgeons (ACS) and in partnership with the US Department of Veterans Affairs (VA), the National Surgical Quality Improvement Program (NSQIP) has been developed to improve the quality of surgical care in adults on a national level. Its purpose is to provide reliable, risk adjusted outcomes data so that surgical quality can be assessed and compared between institutions. Data analysis consists of reporting observed to expected ratios (O/E) for 30-day postoperative mortality and morbidity measurements. A surgical clinical nurse reviewer is assigned at each medical center to collect information on 97 variables, including preoperative, operative, and postoperative factors for patients undergoing major operations in the specialties of general and vascular surgery. Eligible operations are entered into the database on a structured 8-day cycle to ensure representative sampling of cases. Since the introduction of the program into the VA system, there has been a 47% reduction in 30-day postoperative mortality and a 42% reduction in 30-day postoperative morbidity. Over 160 institutions have enrolled with the ACS in its adult NSQIP. In 2005, a planning committee was formed by the ACS and the American Pediatric Surgical Association to explore the development of a children's surgery NSQIP module. In conjunction with the Colorado Health Outcomes Program at the University of Colorado, a program potentially applicable to all children's surgical specialties has been designed. This manuscript describes the development of that Children's ACS-NSQIP module. PMID- 18395664 TI - Outcomes of pediatric anesthesia. AB - Improvement in anesthesia outcomes has derived from advances in safety science related to equipment, drugs, human factors analysis, professional standardization and organization, subspecialty care, and regionalization. Outcomes of pediatric anesthesia have improved, but universal outcome measures are lacking. Because of the limitations of small numbers, future improvement efforts will necessarily involve multiple disciplines, institutions, and regions, and will require sophisticated systems approaches. PMID- 18395666 TI - Bleeding complications in dermatologic surgery. AB - Although the overall incidence is low, bleeding complications in dermatologic surgery can occur and be the source of significant patient morbidity. In this article, we summarize the key aspects of preoperative assessment of patients at risk for bleeding. A review of current issues and literature regarding safe continuation of anticoagulant and antiplatelet medications in dermatologic surgery patients is also presented. In addition, principles for management of bleeding events, should they occur, are also highlighted. PMID- 18395667 TI - Update on botulinum toxin. AB - Botulinum toxin for facial enhancement is currently the most popular aesthetic procedure performed in the United States. New developments have occurred within the last few years. Patients prefer having multiple areas of the upper face treated which increases patient satisfaction. Treatment of the forehead is now being accomplished with fewer units of botulinum toxin. This helps preserve the natural look of some movement of the forehead. Men require more units of botulinum toxin than women. Combination therapy using botulinum toxin along with lasers or filler substances is ideal. Aesthetic medicine knowledge has progressed, contributing a greater understanding of botulinum treatment for advanced areas of the face. The orbicularis oris, mentalis, and depressor anguli oris are now routinely treated and help improve overall facial appearance. Other forms of botulinum toxins (additional type A or type B toxins) are available, each with advantages and disadvantages. PMID- 18395668 TI - What is new in the treatment of steroid-induced osteoporosis? AB - Glucocorticoid-induced osteoporosis (GIOP) is a serious complication resulting from long-term steroid treatment. In addition to several nonpharmacologic therapies recommended by the American College of Rheumatology, various pharmacologic therapies, such as calcium, vitamin D, hormone-replacement therapy, calcitonin, and bisphosphonates, can be used to prevent and/or treat GIOP. Bisphosphonates, which are potent inhibitors of bone resorption, are considered the most effective and first-line agents for increasing bone mineral density and decreasing the risk of fracture. Human parathyroid hormone has emerged as a promising agent for the treatment of severe GIOP when used alone or in combination with a bisphosphonate. PMID- 18395669 TI - Depression and suicidal behavior in acne patients treated with isotretinoin: a systematic review. AB - Isotretinoin (13-cis retinoic acid) is an effective treatment for severe cystic or recalcitrant acne vulgaris; however, concerns have been raised regarding its potential association with depression and suicidal behavior. We sought to explore the proposed relationship between isotretinoin use and the risk of depression and attempted and completed suicide in patients with acne vulgaris by performing a systematic literature search for studies reporting primary data on depression and suicidal behavior in patients treated with isotretinoin for acne vulgaris. Nine studies met the qualifying criteria for our analysis. Rates of depression among isotretinoin users ranged from 1% to 11% across studies, with similar rates in oral antibiotic control groups. Overall, studies comparing depression before and after treatment did not show a statistically significant increase in depression diagnoses or depressive symptoms. Some, in fact, demonstrated a trend toward fewer or less severe depressive symptoms after isotretinoin therapy. This decrease was particularly evident in patients with pretreatment scores in the moderate or clinical depression range. No correlation between isotretinoin use and suicidal behavior was reported, although only one retrospective study presented data on this topic. Although the current literature does not support a causative association between isotretinoin use and depression, there are important limitations to many of the studies. The available data on suicidal behavior during isotretinoin treatment are insufficient to establish a meaningful causative association. PMID- 18395670 TI - Epidermal nevus syndromes. AB - The term "epidermal nevus syndrome" (ENS) has been used to describe the association of epidermal hamartomas and extra-cutaneous abnormalities. Epidermal nevi follow the lines of Blaschko. The majority of the extra-cutaneous manifestations involve the brain, eye, and skeletal systems. Several subsets with characteristic features have been delineated including the nevus sebaceous syndrome, Proteus syndrome, CHILD syndrome, Becker nevus syndrome, nevus comedonicus syndrome, and phakomatosis pigmentokeratotica. Epidermal nevi have been associated with benign and malignant neoplasms. Advances in molecular biology have revealed that the manifestations of ENS are due to genomic mosaicism. It is likely that the varied clinical manifestations of ENS are due in great part to the functional effects of specific genetic defects. Optimal management of the patient with ENS involves an interdisciplinary approach. Amelioration of the cutaneous features of ENS has been difficult but there have been advances, especially in the use of lasers. PMID- 18395671 TI - Congenital melanocytic nevi: treatment modalities and management options. AB - Congenital melanocytic nevi can be cosmetically disfiguring, give rise to melanoma, and suggest the presence of neurocutaneous melanocytosis. Management decisions must be tailored for each patient and each nevus, taking into consideration the risk for developing malignancy, risk for developing symptomatic neurocutaneous melanocytosis, cosmetic implications of having the nevus, cosmetic implications of any resultant surgical scars from their removal, adverse effects that the nevus may have on psycho-social development, and the adverse effects and long-term sequelae of any surgical intervention. The advantages and disadvantages of different modalities used in the treatment of congenital melanocytic nevi are discussed. Organizational flow diagrams are presented to help clinicians in managing patients with different sized congenital melanocytic nevi. PMID- 18395672 TI - The effect of pharmacological stress on intraventricular dyssynchrony in left ventricular systolic dysfunction. AB - BACKGROUND: Cardiac resynchronisation therapy (CRT) improves symptoms and exercise capacity in many patients with heart failure (HF) who have left ventricular systolic dysfunction (LVSD) and markers of dyssynchrony. LV dyssynchrony is conventionally measured at rest but the symptoms of heart failure occur predominantly on exercise. Induction or exacerbation of dyssynchrony during stress might identify additional patients who could benefit from CRT. METHODS AND RESULTS: Seventy-seven patients (47 with QRSd<120 ms and 30 with QRSd>120 ms) with heart failure due to left ventricular systolic dysfunction and 22 normal subjects underwent dobutamine stress echocardiography using colour tissue Doppler imaging. Left intraventricular dyssynchrony was measured as the standard deviation of the time to peak velocity from the onset of the QRS (Ts-SD) and the difference between the maximum and minimum time to peak velocity (Tscor-diff) in the 12 non-apical segments at rest and during peak stress. Timings were corrected for heart rate. The mean values of these indices increased with stress in both groups of patients but not in control subjects (p<0.001). The prevalence of conventionally-defined dyssynchrony also increased with stress. CONCLUSION: In patients with heart failure, the severity and the prevalence of intraventricular dyssynchrony increase with stress. Whether stress-induced dyssynchrony will identify patients who might benefit from CRT awaits further research. PMID- 18395673 TI - Left ventricular pacing site in cardiac resynchronization therapy: clinical follow-up and predictors of failed lateral implant. AB - The effects of the left ventricular (LV) pacing site on the clinical results of resynchronization therapy (CRT) are not well characterized. The aim of this study was to define the effect of LV lead location on clinical response and LV remodelling, and to identify predictors of failure to implant the LV lead in a lateral location. One hundred and seventy two consecutive patients were evaluated at baseline and 6 months after CRT. In 128 patients, the LV lead was implanted in the lateral region (Group 1), while 44 received an anterior implant due to anatomical or electrical factors (Group 2). Group 1 was associated with a significantly better functional outcome assessed both by NYHA class (p<0.001) and by the six-minute-walk test (p=0.01) compared with group 2. LV ejection fraction and volumes, and inter- and intraventricular dyssynchrony only improved significantly (p<0.01) in group 1. The only independent predictor of a failed lateral implant was the presence of ischaemic cardiomyopathy (OR 3.29, 95% CI 2.2 13.7; p=0.02). In conclusion, a lateral lead location results in a better functional outcome and greater reverse LV remodelling compared with anterior locations. The presence of ischaemic cardiomyopathy is a risk factor for a failed lateral LV implant. PMID- 18395674 TI - Sex and sex steroids in cardiovascular-renal physiology and pathophysiology. PMID- 18395676 TI - Estrogen receptors in the kidney: lessons from genetically altered mice. AB - BACKGROUND: Sex differences in human and animal models of kidney disease suggest that estrogen receptor (ER)-mediated events may modulate these processes. Genetically altered mice lacking one or both ERs provide a powerful tool to study these phenomena. OBJECTIVE: This article examines sex differences in the kidney, particularly the role of ERs. METHODS: To identify pertinent studies in genetically altered mice, a literature search was conducted on the MEDLINE database from January 1966 to July 2007, using the search terms estrogen receptor, kidney, and mice. Our group examined the effect of the ER-alpha knockout genotype on the kidney in streptozotocin-induced diabetes mellitus and compensatory kidney growth after uninephrectomy. RESULTS: Female mice lacking ERa had reduced renal growth, including glomerular enlargement after 2 weeks of streptozotocin-induced diabetes mellitus and compensatory kidney growth 48 hours after uninephrectomy. CONCLUSION: ER-mediated events influence kidney growth and disease in female mice. PMID- 18395675 TI - Estrogens and the diabetic kidney. AB - BACKGROUND: Across all ages, the incidence and rate of progression of most nondiabetic renal diseases are markedly higher in men compared with age-matched women. These observations suggest that female sex may be renoprotective. In the setting of diabetes, however, this female protection against the development and progression of renal disease is diminished. OBJECTIVE: This review aimed to summarize our current understanding of sex differences in the development and progression of diabetic renal disease, and of the contribution of sex hormones, particularly estrogens, to the pathophysiology of this disease. We also attempted to answer why female sex does not protect the diabetic kidney. METHODS: Using terms such as gender, sex, diabetes, diabetic nephropathy, estrogens, and sex hormones, the PubMed database was searched for English-language articles; targeted searches were conducted using terms such as gender/sex differences in diabetic renal disease. No restrictions were imposed on publication dates. RESULTS: Although the existing data regarding the sex differences in the incidence and progression of diabetic renal disease are inconclusive, the undisputed fact is that women with either type 1 or type 2 diabetes mellitus exhibit a much higher incidence of renal disease compared with nondiabetic women. It is conceivable that the loss of female sex as a renoprotective factor in diabetes may be related to the abnormal regulation of sex hormone concentrations. Both clinical and experimental data suggest that diabetes may be associated with an imbalance in estradiol concentrations. Supplementation with 17beta-estradiol or administration of selective estrogen receptor modulators reduces the incidence of diabetes and attenuates the progression of diabetic renal disease. CONCLUSIONS: Serum concentrations of ovarian hormones may provide a new means for predicting future risk of renal complications in diabetes. Exogenous steroid hormones may be an effective treatment for attenuating the progression of diabetic nephropathy. PMID- 18395677 TI - Androgens stimulate proximal tubule transport. AB - BACKGROUND: Disrupting the enzyme cytochrome P4a14 in mice leads to hypertension, which is more severe in male than in female mice and appears to be due to androgen excess. Androgens are known to increase expression of angiotensinogen,but the effect of androgens on proximal tubule transport is unknown. OBJECTIVE: These studies aimed to determine the effect of androgens on proximal tubule transport. METHODS: Proximal tubules from knockout (KKO) and wild ttype (WWT) (SSV/1129) mice were perfused in vitro. Volume resorption (JJ v ) was measured using 3 H-methoxy inulin as a volume marker. In separate experiments, male Sprague-Dawley rats were given dihydrotestosterone (DDHT) injections IP for 10 days. Proximal tubule transport was measured in this model using in vivo microperfusion. The renal expression of angiotensinogen was measured by Northern analysis, and brush border membrane protein abundance of the sodium-hhydrogen exchanger isoform 3 (NNHE3) was measured by Western blotting in the control and DHT-ttreated rats. RESULTS: Mean (SSE) Jv was significantly elevated in proximal tubules from KO mice compared with WT mice (11.11 [0.006] vs 0.77 [0.112] nL/mm . mm, respectively; P<0.05). The mean proximal tubule Jv rate was significantly higher in DHT-ttreated rats than in control rats given vehicle injections (44.57 [0.331] vs 3.31 [0.223] nL/mm . min, respectively; P<0.01). Luminal perfusion with either enalaprilat or losartan decreased the proximal tubule J v rate in DHT ttreated rats to a greater degree than in control rats. The DHT-treated rats had higher blood pressures and lower serum angiotensin II concentrations than did the control rats. CONCLUSION: Results suggest that androgens may directly upregulate the proximal tubule reninangiotensin system, increase the expression of NHE3, and increase the Jv rate, thereby increasing extracel-lular volume and blood pressure and secondarily decreasing serum angiotensin II concentrations. PMID- 18395678 TI - Sex differences in the fetal programming of hypertension. AB - BACKGROUND: Numerous clinical and experimental studies support the hypothesis that the intrauterine environment is an important determinant of cardiovascular disease and hypertension. OBJECTIVE: This review examined the mechanisms linking an adverse fetal environment and increased risk for chronic disease in adulthood with an emphasis on gender differences and the role of sex hormones in mediating sexual dimorphism in response to a suboptimal fetal environment. METHODS: This review focuses on current findings from the PubMed database regarding animal models of fetal programming of hypertension, sex differences in phenotypic outcomes, and potential mechanisms in offspring of mothers exposed to an adverse insult during gestation. For the years 1988 to 2007, the database was searched using the following terms: fetal programming, intrauterine growth restriction, low birth weight, sex differences, estradiol, testosterone, high blood pressure, and hypertension. RESULTS: The mechanisms involved in the fetal programming of adult disease are multifactorial and include alterations in the regulatory systems affecting the long-tterm control of arterial pressure. Sex differences have been observed in animal models of fetal programming, and recent studies suggest that sex hormones may modulate activity of regulatory systems, leading to a lower incidence of hypertension and vascular dysfunction in females compared with males. CONCLUSIONS: Animal models of fetal programming provide critical support for the inverse relationship between birth weight and blood pressure. Experimental models demonstrate that sex differences are observed in the pathophysiologic response to an adverse fetal environment. A role for sex hormone involvement is strongly suggested,with modulation of the renin-angiotensin system as a possible mechanism. PMID- 18395679 TI - Pathophysiology of hypertension in response to placental ischemia during pregnancy: a central role for endothelin? AB - BACKGROUND: Preeclampsia is new-onset hypertension with proteinuria during pregnancy. The initiating event in preeclampsia has been postulated to involve reduced placental perfusion, which leads to widespread dysfunction of the maternal vascular endothelium. OBJECTIVE: The main objective of this brief review was to highlight some of the recent advances in our understanding of the mechanisms whereby the endothelin (ET) system, via ET type A (ETA) receptor activation, modulates blood pressure in preeclamptic women and in animal models of pregnancy-related hypertension. METHODS: This review focused on the role of ET and tumor necrosis factor-alpha (TNF-alpha) in preeclampsia, with emphasis on the pathophysiology of hypertension in response to placental ischemia in animal models of pregnancy. Relevant published data were identified by searching PubMed and supplemented with contributions from our laboratory. RESULTS: Studies in preeclamptic women indicate that their hypertension is associated with increases in ET synthesis. Recent studies in pregnant rats indicate that the ET system is activated in response to reductions in uterine perfusion pressure and to chronic elevations in serum TNF-alpha concentrations. In these 2 animal models, the findings also suggest that ET A receptor activation may play a role in mediating hypertension. CONCLUSIONS: Although recent studies in animal models implicate an important role for the ET system in preeclampsia, the usefulness of selective ET A receptor antagonists for the treatment of hypertension in women with preeclampsia remains unclear. This important question will not be answered until well-controlled clinical studies using specific ET A receptor antagonists are conducted for women with preeclampsia. PMID- 18395680 TI - Need for research on estrogen receptor function: importance for postmenopausal hormone therapy and atherosclerosis. AB - BACKGROUND: Cardiovascular disease is the leading cause of morbidity and mortality in men and women worldwide. Although rare in premenopausal women, its incidence rises sharply after menopause, indicating atheroprotective effects of endogenous estrogens. OBJECTIVE: This review discusses the differential effects of estrogen receptor function on atherosclerosis progression in pre- and postmenopausal women, including aspects of gender differences in vascular physiology of estrogens and androgens. METHODS: Recent advances in the understanding of the pathogenesis of atherosclerosis, estrogen receptor function, and hormone therapy are reviewed, with particular emphasis on clinical and molecular issues. RESULTS: Whether hormone therapy can improve cardiovascular health in postmenopausal women remains controversial. Current evidence suggests that the vascular effects of estrogen are affected by the stage of reproductive life, the time since menopause, and the extent of subclinical atherosclerosis. The mechanisms of vascular responsiveness to sex steroids during different stages of atherosclerosis development remain poorly understood in women and men. CONCLUSION: In view of the expected increase in the prevalence of atherosclerotic vascular disease worldwide due to population aging, research is needed to determine the vascular mechanism of endogenous and exogenous sex steroids in patients with atherosclerosis. Such research may help to define new strategies to improve cardiovascular health in women and possibly also in men. PMID- 18395681 TI - Gender and human chronic renal disease. AB - BACKGROUND: Gender affects the incidence, prevalence, and progression of renal disease. In animal models of the disease, female sex appears to modify the course of progression. Hormonal manipulation by male or female castration also changes the course of renal disease progression, suggesting direct effects of sex hormones in influencing the course of these maladies. OBJECTIVE: This review examines the pertinent animal and human studies assessing the role of gender, and strives to shed light on the possible physiologic mechanisms underlying the effect of gender, on renal disease progression. METHODS: A summary and evaluation of past and recent studies describing the rate of renal disease progression in animal models and humans as it pertains to gender is provided. In addition, studies elucidating the factors involved in the more modest renal progression rate in females are reviewed and conclusions drawn. Relevant English-language publications were identified by searching the PubMed database from January 1990 until November 2007 using the search terms gender, sex, renal disease, and kidney. RESULTS: In polycystic kidney disease, membranous nephropathy, immunoglobulin A nephropathy, and "chronic renal disease of unknown etiology," men progress at a faster rate to end-stage renal failure than do women. In type 1 diabetes mellitus, there is evidence that males are more likely to manifest signs of renal disease, such as proteinuria. The factors involved in this gender disparity may include diet, kidney and glomerular size, differences in glomerular hemodynamics, and the direct effects of sex hormones. In many, but not all, animal models of renal disease, estrogens slow progression rate. Several studies have recently evaluated the effect of selective estrogen receptor modulators on renal function in humans. CONCLUSION: Further studies assessing the factors involved in the gender disparity in renal disease progression and the effects of hormonal treatments are warranted. PMID- 18395682 TI - Genetic and pharmacologic strategies to determine the function of estrogen receptor alpha and estrogen receptor beta in cardiovascular system. AB - BACKGROUND: The biological functions of estrogens extend beyond the female and male reproductive tract, affecting the cardiovascular and renal systems. Traditional views on the role of postmenopausal hormone therapy (HT) in protecting against heart disease, which were challenged by clinical end point studies that found adverse effects of combined HT, are now being replaced by more differentiated concepts suggesting a beneficial role of early and unopposed HT that does not include a progestin. OBJECTIVE: We reviewed recent insights, concepts, and research results on the biology of both estrogen receptor (ER) subtypes, ERalpha and ERbeta, in cardiac and vascular tissues. Knowledge of these ER subtypes is crucial to understanding gender and estrogen effects and to developing novel, exciting strategies that may have a profound clinical impact. METHODS: This review focuses on in vivo studies and includes data presented at the August 2007 meeting of the American Physiological Society as well as data from a search of the MEDLINE and Ovid databases from January 1986 to November 2007. Search results were restricted to English-language publications, using the following search terms: estrogen, estrogen receptor alpha, estrogen receptor beta, estrogen receptor alpha agonist, estrogen receptor alpha antagonist, estrogen receptor beta agonist, estrogen receptor beta antagonist, PPT, DPN, heart, vasculature, ERKO mice, BERKO mice, transgenic mice, and knockout mice. RESULTS: Genetic mouse models and pharmacologic studies that employed selective as well as nonselective ER agonists support the concept that both ER subtypes confer protective effects in experimental models of human heart disease, including hypertension, cardiac hypertrophy, and chronic heart failure. CONCLUSIONS: Genetic models and novel ligands hold the promise of further improving our understanding of estrogen action in multiple tissues and organs. These efforts will ultimately enhance the safety and efficacy of HT and may also result in new applications for synthetic female sex hormone analogues. PMID- 18395683 TI - Sex steroids and vascular responses in hypertension and aging. AB - BACKGROUND: Sex hormones play a significant role in human physiology. Estrogen may have protective effects in the cardiovascular system, as evidenced by the decreased incidence of cardiovascular disease (CVD) in premenopausal compared with postmenopausal women. OBJECTIVE: This review highlights the acute and long term effects of sex hormones on the vascular endothelium and vascular smooth muscle (VSM) in adults. Changes in the sex hormone mix, their receptors, and their effects on vascular function in hypertension and aging are also discussed. METHODS: Literature collected from the National Centers for Biotechnology Information as identified by a PubMed database search, as well as our experimental work, was used to highlight current knowledge regarding vascular responses to sex hormones in hypertension and in aging. RESULTS: Experiments in adult female animals have shown that estrogen induces endothelium-dependent vascular relaxation via the nitric oxide (NO), prostacyclin, and hyperpolarization pathways. Also, surface membrane estrogen receptors (ERs) decrease intracellular free Ca2+ concentration and perhaps protein kinase C dependent VSM contraction. However, clinical trials such as the Heart and Estrogen/progestin Replacement Study (HERS), HERS-II, and the Women's Health Initiative did not support the experimental findings and demonstrated adverse cardiovascular events of hormone therapy (HT) in aging women. The lack of vascular benefits of HT may be related to the hormone used, the ER, or the patient's cardiovascular condition or age. Experiments on vascular strips from aging (16-month-old) female spontaneously hypertensive rats have shown reduced ER mediated NO production from endothelial cells and decreased inhibitory effects of estrogen on Ca2+ entry mechanisms of VSM contraction. The age-related decrease in ER-mediated vascular relaxation may explain the decreased effectiveness of HT on CVD in aging women. CONCLUSIONS: New HT strategies should further examine the benefits of natural estrogens and phytoestrogens. Transdermal estrogen may be more effective than the oral form, and specific ER modulators may maximize the vascular benefits and reduce the risk of invasive breast cancer. Variants of vascular ERs should be screened for genetic polymorphisms and postmenopausal decrease in the amount of downstream signaling mechanisms. HT may be more effective during the menopausal transition than in late menopause. Progesterone, testosterone, or their specific modulators may be combined with estrogen to provide alternative HT strategies. Thus, HT type, dose, route of administration, and timing should be customized, depending on the patient's cardiovascular condition and age, thereby enhancing the vascular benefits of HT in aging women. PMID- 18395684 TI - Differential effects of sex steroids in young and aged female mRen2.Lewis rats: a model of estrogen and salt-sensitive hypertension. AB - BACKGROUND: Male-female differences in the expression of hypertension and in end organ damage are evident in both experimental models and human subjects, with males exhibiting a more rapid onset of cardiovascular disease and mortality than do females. The basis for these male-female differences is probably the balance of the complex effects of sex steroids (androgens, estrogen, progesterone) and their metabolites on the multiple regulatory systems that influence blood pressure (BP). A key target of estrogen and other steroids is likely to be the different components of the renin-angiotensin-aldosterone system (RAAS). OBJECTIVE: The aim of this study was to review the current experimental evidence on the protective effects of estrogen in hypertensive models. METHODS: The search terms estrogen , renin-aangiotensin-aldosterone system, renin receptor, salt sensitivity, endorgan damage, hypertension, kidney, mRen2. Lewis, and injury markers were used to identify relevant publications in the PubMed database (restricted to the English language) from January 1990 to October 2007. RESULTS: In a new congenic model that expresses the mouse renin 2 gene (mRen2. Lewis), estrogen depletion (via ovariectomy [OVX ]) in young rats was found to have a marked stimulatory effect on the progression of increased BP and cardiac dysfunction. Moreover, estrogen depletion exacerbated salt-sensitive hypertension and the extent of salt-induced cardiac and renal injury in young mRen2. Lewis rats, which probably reflected the inability to appropriately regulate various components of the RAAS. However, OVX in aged mRen2. Lewis rats conveyed renal protective effects from a high-salt diet compared with intact hypertensive littermates (64 weeks), and these effects were independent of changes in BP. CONCLUSION: These studies in hypertensive mRen2. Lewis rats underscored the influence of ovarian hormones on BP and tissue injury, as well as the plasticity of this response, apparently due to age and salt status. PMID- 18395686 TI - Estrogen, inflammation, and platelet phenotype. AB - BACKGROUND: Although exogenous estrogenic therapies increase the risk of thrombosis, the effects of estrogen on formed elements of blood are uncertain. OBJECTIVE: This article examines the genomic and nongenomic actions of estrogen on platelet phenotype that may contribute to increased thrombotic risk. METHODS: To determine aggregation, secretion, protein expression, and thrombin generation, platelets were collected from experimental animals of varying hormonal status and from women enrolled in the Kronos Early Estrogen Prevention Study. RESULTS: Estrogen receptor beta predominates in circulating platelets. Estrogenic treatment in ovariectomized animals decreased platelet aggregation and adenosine triphosphate (ATP) secretion. However, acute exposure to 17beta-estradiol did not reverse decreases in platelet ATP secretion invoked by lipopolysaccharide. Thrombin generation was positively correlated to the number of circulating microvesicles expressing phosphatidylserine. CONCLUSION: Assessing the effect of estrogen treatments on blood platelets may lead to new ways of identifying women at risk for adverse thrombotic events with such therapies. PMID- 18395687 TI - Newborn weight charts underestimate the incidence of low birthweight in preterm infants. AB - OBJECTIVE: The objective of the study was to compare sonographic fetal weight estimates with newborn weight charts and analyze the predictive accuracy of the ponderal index (PI) in preterm infants. STUDY DESIGN: We generated sonographic reference curves for fetal weight and PI estimates from a database of fetal biometric records from 12,589 term deliveries. We then plotted sonographic and newborn weight and PI of 2406 preterm newborns on these curves and compared them with published newborn weight charts. RESULTS: The third centiles of sonographic and newborn weights diverged markedly between 25 and 36 weeks of gestation and by more than 400 g at 32-33 weeks. In contrast, sonographic and newborn PI values were similar despite uncertainties as to fetal length. CONCLUSION: We suggest using sonographic reference fetal weights to screen preterm newborns for low birthweight. Uncertainties in fetal length threaten the reliability of the PI. PMID- 18395685 TI - Effects of botanical dietary supplements on cardiovascular, cognitive, and metabolic function in males and females. AB - BACKGROUND: The onset of menopause marks a pivotal time in which the incidence of hypertension and of cardiovascular disease (CVD) begins to increase dramatically in women. Before menopause, the incidences of these diseases are significantly lower in women than in age-matched men. After menopause, the rates of these diseases in women eventually approximate those in men. The loss of endogenous estrogen at menopause has been traditionally believed to be the primary factor involved in these changes. OBJECTIVE: This review summarizes recent findings regarding the effectiveness of botanicals in the treatment of some menopausal symptoms and other symptoms of aging (eg, rise in arterial pressure, cognitive decline, insulin resistance, and hyperlipidemia). METHODS: Articles were selected for inclusion in this review based on the significance of the research and contribution to the current understanding of how each botanical elicits cardioprotective effects. To this end, PubMed and MEDLINE databases were searched, using terms that included the name of the specific botanical along with the relevant aspects of its action(s), such as blood pressure, glycemic control, and lipids. Most of the articles used were published within the past 5 years, although some older articles that were seminal in advancing the current understanding of botanicals were also included. RESULTS: Soy has been found to lower plasma lipid concentrations and arterial pressure in postmenopausal women and age-matched men, and to have protective effects in heart disease and atherosclerosis of the carotid and coronary circulation. Soy was also found to lower fasting insulin concentrations and glycosylated hemoglobin concentrations. Grape seed extract, another frequently used botanical, contains polyphenols that have been found to reduce arterial pressure and salt-sensitive hypertension in estrogendepleted animal models. CONCLUSION: Several botanical compounds have been found to have beneficial effects in the treatment of the symptoms of menopause and other symptoms of aging, including CVD, cognitive decline, and metabolic diseases. PMID- 18395688 TI - Reducing high-order perineal laceration during operative vaginal delivery. AB - OBJECTIVE: This study was undertaken to assess the impact of a focused intervention on reducing high-order (third and fourth degree) perineal lacerations during operative vaginal delivery. STUDY DESIGN: The following recommendations for clinical management were promulgated by departmental lectures, distribution of pertinent articles and manuals, training of physicians, and prominent display of an instructional poster: (1) increased utilization of vacuum extraction over forceps delivery; (2) conversion of occiput posterior to anterior positions before delivery; (3) performance of mediolateral episiotomy if episiotomy was deemed necessary; (4) flexion of the fetal head and maintenance of axis traction; (5) early disarticulation of forceps; and (6) reduced maternal effort at expulsion. Peer comparison was encouraged by provision of individual and departmental statistics. Clinical data were extracted from the labor and delivery database and the medical record. RESULTS: One hundred fifteen operative vaginal deliveries occurred in the 3 quarters preceding the intervention, compared with 100 afterward (P = .36). High-order laceration with operative vaginal delivery declined from 41% to 26% (P = .02), coincident with increased use of vacuum (16% vs 29% of operative vaginal deliveries, P = .02); fewer high order lacerations after episiotomy (63% vs 22%, P = .003); a nonsignificant reduction in performance of episiotomy (30% vs 23%, P = .22); and a nonsignificant increase in mediolateral episiotomy (14% vs 30% of episiotomies, P = .19). CONCLUSION: Introduction of formal practice recommendations and performance review was associated with diminished high-order perineal injury with operative vaginal delivery. PMID- 18395689 TI - Feasibility of platinum-based chemotherapy during pregnancy. PMID- 18395690 TI - Follow-up in obese pregnant women to prevent stillbirth. PMID- 18395691 TI - Hormones restore biomechanical properties of the vagina and supportive tissues after surgical menopause in young rats. AB - OBJECTIVE: The objective of the study was to determine the impact of hormones on the biomechanical properties of the vagina and its supportive tissues following surgical menopause in young vs middle-aged rats. STUDY DESIGN: Long-Evans rats (4 month virgin [n = 34], 4-month parous [n = 36], and 9-month parous [n = 34]), underwent ovariectomy (OVX) or sham surgery. OVX animals received hormones (estrogen [E2] or estrogen plus progesterone [E2 plus P4]), placebo, or a matrix metalloproteinase inhibitor (chemically modified tetracycline-8 [CMT-8]). Animals were euthanized after 8 weeks and the biomechanical properties of the vagina and supportive tissues determined. Data were analyzed using a 1-way analysis of variance and posthoc tests. RESULTS: OVX induced a rapid decline in the biomechanical properties of pelvic tissues in young but not middle-aged rats. Supplementation with E2, E2 plus P4, or CMT-8 restored tissues of young rats to control levels with no effect on middle-aged tissues. Parity did not have an impact on tissue behavior. CONCLUSION: OVX has a differential effect on the tissues of young vs middle-aged rats. PMID- 18395692 TI - Tailored intervention to increase dual-contraceptive method use: a randomized trial to reduce unintended pregnancies and sexually transmitted infections. AB - OBJECTIVE: The objective of the study was to determine whether a transtheoretical model-tailored expert system intervention increases dual-method contraceptive use, compared with a nontailored educational intervention. STUDY DESIGN: We performed a randomized clinical trial of 542 women at high risk for sexually transmitted infections (STIs) and unintended pregnancy. The intervention group received computer-based tailored feedback using a multimedia program. The control group received general contraceptive information and nontailored advice. RESULTS: Participants in the intervention group were more likely to report use of dual contraceptive methods during follow-up (adjusted hazard rate ratio 1.70, 95% confidence interval 1.09, 2.66), compared with controls. However, there were no differences in the rates of incident STI or unintended pregnancy between the 2 groups. CONCLUSIONS: The computer-based transtheoretical model-tailored intervention resulted in a 70% increase in reported dual-method contraceptive use in a group of women at high risk for STIs and unintended pregnancy. Inconsistent use of dual methods may explain the lack of effect on unintended pregnancy rates and incident STIs. PMID- 18395693 TI - Clostridium difficile-associated diarrhea: an emerging threat to pregnant women. AB - OBJECTIVE: To estimate if Clostridium difficile-associated disease (CDAD) is increasing in peripartum women. STUDY DESIGN: Peripartum CDAD was assessed through 1) passive surveillance collecting clinical and pathology data on severe cases and 2) survey among infectious disease consultants (ICDs) in the Emerging Infections Network. RESULTS: Ten severe cases were collected; most had associated antibiotic use. Seven women were either admitted to the ICU or underwent colectomy. Three infants were stillborn, and 3 women died. The epidemic Clostridium difficile strain was found in 2 cases. Among 798 ICDs, 419 (52%) participated in the survey. Thirty-seven respondents (9%) recalled 55 cases, mostly in the postpartum period with 21 complications, mainly due to relapse. CONCLUSION: Severe CDAD may be increasing in peripartum women. Clinicians should have a low threshold for testing, be aware of the potential for severe outcomes, and take steps to reduce both the risk of disease and resultant complications. PMID- 18395694 TI - The sleep and performance of train drivers during an extended freight-haul operation. AB - The sleep and performance of train drivers was monitored across a 106-h rail operation between the Australian cities of Adelaide and Perth. The drivers worked alternating 8-h shift rotations across the operation and rested in specially equipped, crew-van carriages during non-work periods. The crew-van rest opportunities were associated with shorter bedtime spans, less total sleep time, and poorer sleep efficiency than sleeps initiated at home. The duration of crew van sleeps was primarily dependent on the time of day at which the rest opportunities occurred. Overall, drivers incurred a significant cumulative sleep loss across the duration of the operation. Despite the deficit, drivers were able to sustain vigilance performance across the operation. PMID- 18395695 TI - Key elements on team achievement: a retrospective analysis. AB - This study is a historical reconstruction and in-depth scrutiny of one very successful team initiative within a Quality Improvement (QI) program in a municipal government organization. The identification of the essential concepts that contributed to the team's success and their integration into a local explanatory theory of team achievement is the final purpose of the study. The focus of this retrospective study is on the team process, with primary attention given to team members' perspectives and reflections on the project development. Data collection and analysis were conducted with methods drawn from the qualitative research tradition and from Quality Management. Main findings point to the importance of consistent management support, correct team composition with an emphasis on the team leader choice and demeanor, and to the central role of training in the group conduct. The negotiated approach to decision-making employed by the team, which reflected its organizational context, and the effort and mechanisms that allowed the group to reach equilibrium between internal and external interests proved critical for its ultimate achievement. PMID- 18395696 TI - Interplay of mechanical loading and growth factors in the mandibular condyle. AB - The mandibular condyle is an important growth site in the developing mandible. The growth of the condyle is known to be highly adaptable to functional factors. This property is exploited in orthodontics for the treatment of class II malocclusions and mandibular asymmetries. However, there is an ongoing debate on the efficacy of functional appliances. The comparison of experimental studies is complicated by the lack of detailed analyses of the load distribution within the condyle. In spite of this, there is a large body of evidence showing that mechanical manipulation of the condyle induces metabolic changes, and changes in the expression of growth factors and other signalling molecules. This review aims to give an overview of the role of growth factors in the condyle with special emphasis on their responsiveness to mechanical perturbation. PMID- 18395697 TI - Effects of commonly used food preservatives on biofilm formation of Streptococcus mutans in vitro. AB - OBJECTIVE: Sodium benzoate (SB), potassium sorbate (PS) and sodium nitrite (SN) are commonly used food preservatives. In this in vitro study, the effects of these substances on biofilm formation of Streptococcus mutans were analysed. METHODS: In addition to the microtiter plate test (MPT), a biofilm reactor containing bovine enamel slabs (BES) was used to study the influence of food preservatives on biofilm formation in 5 independent periods of 4 days each. These included one period with chlorhexidine digluconate (CHX) as a positive control as well as a period with growth medium alone as a negative control. The vitality of the biofilm on BES was detected using live/dead staining and confocal laser scanning microscopy. Additionally, the number of colony forming units (CFU) was determined. RESULTS: In MPT 0.12% SN significantly reduced the biofilm formation. PS at a concentration of 0.4% tended to inhibit biofilm formation, whereas the inhibition for 0.8% PS was significant. Less inhibition was caused by 0.8% SB. In the biofilm reactor 0.06% of SN, 0.1% of SB and 0.1% PS significantly reduced the covering grade as well as the CFU of the biofilm. Biofilm vitality was reduced significantly by CHX to a level of 32.5% compared to the control. Only SB reduced the vitality to a level of 19.1%. SN and PS showed no influence on biofilm vitality. CONCLUSION: This study indicates the potential of food preservatives as inhibitory agents in S. mutans biofilm formation, which should be kept in mind when studying the effects of conserved food on dental plaque biofilm in situ. PMID- 18395698 TI - The effect of Streptococcus mutans and Candida glabrata on Candida albicans biofilms formed on different surfaces. AB - Although Candida containing biofilms contribute to the development of oral candidosis, the characteristics of multi-species Candida biofilms and how oral bacteria modulate these biofilms is poorly understood. The aim of this study was to investigate interactions between Candida albicans and either Candida glabrata or Streptococcus mutans in biofilms grown on various surfaces, with or without saliva. Hydroxyapatite (HA), polymethylmetacrylate (PMMA) and soft denture liner (SL) discs were used as substratum. Counts of viable micro-organisms in the accumulating biofilm layer were determined and converted to colony forming units per unit surface area. Confocal laser scanning microscopy was used to characterize biofilms and to quantitate the number of hyphae in each condition tested. Viable counts of C. albicans and C. glabrata per mm(2) decreased in the order HA>PMMA>SL (p<0.05). Biofilms grown on saliva-coated specimens harboured fewer C. glabrata than uncoated specimens (p<0.05). Glucose and the presence of S. mutans suppressed C. albicans hyphal formation. Dual Candida species biofilms did not show competitive interaction between the two species. We conclude that Candida biofilms are significantly affected by saliva, substratum type and by the presence of other micro-organisms. PMID- 18395699 TI - Cerebrospinal fluid biomarkers in Parkinson's disease with dementia and dementia with Lewy bodies. AB - BACKGROUND: Clinical criteria for differentiating Parkinson's disease (PD) with dementia (PDD) from dementia with Lewy bodies (DLB) are unsatisfactory. Their existence as distinct clinicopathologic entities is still debated, although the burden of Alzheimer's disease (AD) pathology seems higher in DLB. Thus, analysis of cerebrospinal fluid (CSF) biomarkers (beta-amyloid(1-42) [Abeta42], total tau, and hyperphosphorylated tau [p-tau]) in living subjects might provide significant pathophysiological information on these diseases. METHODS: Cerebrospinal fluid biomarkers were measured in DLB (n = 19), PDD (n = 18), and AD (n = 23) subjects matched for age, sex, and dementia severity, as well as in PD (n = 20) and normal control subjects (n = 20). RESULTS: DLB showed the lowest mean CSF Abeta42 levels, with a negative association to dementia duration (rho = -.42, p = .07). In DLB patients, mean CSF total tau levels were significantly lower than in AD patients (508 +/- 387 vs. 960 +/- 619, respectively) but twofold to threefold higher than in PDD (286 +/- 184), PD (160 +/- 64), or normal control subjects (177 +/- 76), with a positive association to dementia severity (Mini-Mental State Examination: rho = -.54, p = .02; Milan Overall Dementia Assessment: rho = -.66, p = .002). PDD patients had mean CSF Abeta42 and total tau levels similar to those seen in PD patients. Hyperphosphorylated tau was significantly increased in the AD group only. CONCLUSIONS: Cerebrospinal fluid Abeta42 and total tau have a different behavior in DLB and PDD, being related to duration and severity of dementia in DLB alone. Hyperphosphorylated tau is not significantly altered in these conditions. PMID- 18395700 TI - Sensory gating in schizophrenia: P50 and N100 gating in antipsychotic-free subjects at risk, first-episode, and chronic patients. AB - BACKGROUND: Abnormal sensory gating in schizophrenia has frequently been reported; however, only limited data on unmedicated patients and patients at risk to develop a psychosis have, as yet, been available. METHODS: P50 and N100 suppression were assessed with an auditory double-click paradigm in five groups: 18 at-risk subjects who did not develop a full psychosis within the follow-up period of 2 years, 21 truly prodromal subjects who developed frank psychosis within the follow-up period, 46 antipsychotic-naive subjects with first-episode schizophrenia, 20 antipsychotic-free subjects with chronic schizophrenia, and 46 healthy control subjects. RESULTS: P50 and N100 suppression indices differed significantly between groups and were lowest in chronic schizophrenia patients. Compared with healthy control subjects, P50 suppression was significantly impaired in at-risk subjects, truly prodromal and first-episode patients (stimulus 2 [S2]/stimulus 1 [S1] P50 amplitude ratio), and chronic schizophrenia patients (difference and ratio), and N100 suppression was significantly reduced in truly prodromal and first-episode patients (S1-S2 difference) and in chronic schizophrenia patients (difference and ratio) but not at-risk subjects. At-risk subjects with and without conversion to psychosis did not significantly differ on any test parameter. CONCLUSIONS: Sensory gating is already impaired in early stages of schizophrenia, though this is most prominent in chronic stages. Future studies will have to clarify the type and impact of variables modifying sensory gating disturbances, such as illness progression and genetic load. Furthermore, the meaning and nature of differences between P50 and N100 suppression need further elucidation. PMID- 18395701 TI - The role of the cerebellum in schizophrenia. AB - For many years the cerebellum has been considered to serve as a coordinator of motor function. Likewise, for many years schizophrenia has been considered to be a disease that primarily affects the cerebrum. This review summarizes recent evidence that both these views must be revised in the light of emerging evidence about cerebellar function and the mechanisms of schizophrenia. Evidence indicating that the cerebellum plays a role in higher cortical functions is summarized. Evidence indicating that cerebellar abnormalities occur in schizophrenia is also reviewed. These suggest interesting directions for future research. PMID- 18395702 TI - Modafinil modulates GABA-activated currents in rat hippocampal pyramidal neurons. AB - Modafinil is a well-known psychoactive drug used to treat narcolepsy, hypoglycemia, cerebral ischemia and Parkinson's disease. Previous studies showed that ATP-sensitive potassium channels (K(ATP)) play a key role in response to cerebral ischemia, hypoglycemia or metabolic inhibition. Modafinil (0.01-1 mM) dose-dependently decreased the GABA-activated currents (I(GABA)). Pretreatment with the K(ATP) channel blocker, glibenclamide (10 microM), significantly reduced the decrease of I(GABA) caused by modafinil. Thus, the inhibitory effect of modafinil on the I(GABA) is indirect by modulating K(ATP) channel activation, at least in part mediated by K(ATP) channel. PMID- 18395704 TI - 2D selective-TOCSY-DQFCOSY and HSQC-TOCSY NMR experiments for assignment of a homogeneous asparagine-linked triantennary complex type undecasaccharide. AB - The assignment of (1)H and (13)C NMR signals of a complex type triantennary asialooligosaccharide was examined using 2D selective-TOCSY-DQFCOSY and HSQC TOCSY experiments. The 2D selective-TOCSY-DQFCOSY experiment exhibits a 2D DQFCOSY spectrum of an individual monosaccharide in the undecasaccharide, although the NMR signals of several monosaccharides in the triantennary undecasaccharide are heavily overlapped. Selective excitation of each anomeric proton signal and subsequent TOCSY experiment afforded transverse magnetization corresponding to all of the proton signals of the monosaccharide. This magnetization was then developed with the corresponding DQFCOSY pulse sequence to afford the DQFCOSY spectrum of the individual monosugars. In this case, four GlcNAc-b, -e, -j, and -h residues were excited as a mixture. In order to assign (13)C signals, a conventional 2D HSQC-TOCSY spectrum was examined and compared with an unambiguous assignment of 2D selective-TOCSY-DQFCOSY thus obtained. This systematic analysis made it possible to obtain an assignment of the (1)H and (13)C NMR signals of the triantennary undecasaccharide. In addition, these experiments also revealed all of the glycosyl positions in the triantennary undecasaccharide. PMID- 18395703 TI - CRBL cells: establishment, characterization and susceptibility to prion infection. AB - The cerebellum is involved in complex physiological functions including motor control, sensory perception, cognition, language, and emotion. Humans and animals with prion diseases are characterized clinically by ataxia, postural abnormalities and cognitive decline. Pathology in the cerebellum affected by prions includes spongiform degeneration, neuronal loss, and gliosis. To develop an in vitro model system for studying prion biology in cerebellar cells, we established and characterized an immortal cell line (CRBL) isolated from the cerebellum of mice lacking expression of a protein involved in cell cycle arrest. The characteristics of the cells include morphological heterogeneity, rapid proliferation, serum responsiveness during growth, and a change in the number of chromosomes. CRBL cells expressed both neuronal and glial cell markers as well as a considerable level of cellular prion protein, PrP(C). Upon in vitro infection, CRBL cells exhibited selective susceptibility to prions isolated from different sources. These cells chronically propagated prions from SMB cells. Strain specific prion infection in CRBL cells was not due to instability of the cell line, allelic variance, or mutations in the PrP gene. Molecular properties of prions derived from SMB cells were maintained in the infected CRBL cells. Our results suggest that the specific interaction between a prion strain and hosts determined the selective susceptibility of CRBL cells, which reflects the conditions in vivo. In addition to the future studies revealing cellular and molecular mechanism involved in prion pathogenesis, CRBL cells will contribute to the studies dealing with prion strain properties and host susceptibilities. PMID- 18395705 TI - Does language guide event perception? Evidence from eye movements. AB - Languages differ in how they encode motion. When describing bounded motion, English speakers typically use verbs that convey information about manner (e.g., slide, skip, walk) rather than path (e.g., approach, ascend), whereas Greek speakers do the opposite. We investigated whether this strong cross-language difference influences how people allocate attention during motion perception. We compared eye movements from Greek and English speakers as they viewed motion events while (a) preparing verbal descriptions or (b) memorizing the events. During the verbal description task, speakers' eyes rapidly focused on the event components typically encoded in their native language, generating significant cross-language differences even during the first second of motion onset. However, when freely inspecting ongoing events, as in the memorization task, people allocated attention similarly regardless of the language they speak. Differences between language groups arose only after the motion stopped, such that participants spontaneously studied those aspects of the scene that their language does not routinely encode in verbs. These findings offer a novel perspective on the relation between language and perceptual/cognitive processes. They indicate that attention allocation during event perception is not affected by the perceiver's native language; effects of language arise only when linguistic forms are recruited to achieve the task, such as when committing facts to memory. PMID- 18395706 TI - Requirement of Oct3/4 function for germ cell specification. AB - In mammalian embryos, PGCs (primordial germ cells) are specified from a pluripotent epiblast cell population after implantation. In this study, we demonstrated an essential role for the germline-specific transcription factor Oct3/4 in PGC specification. We generated chimeric embryos with ZHBTc4 ES cells lacking both alleles of the Oct3/4 gene (pou5f1). Pluripotency was maintained by an Oct3/4 transgene, and its expression was suppressed by doxycycline (Dox). Transcription of the Oct3/4 transgene in the ES-derived cells unexpectedly suffered constitutive suppression in chimeric embryos without Dox, and the ES derived cells contributed to PGC precursor-like cells, but failed to form PGCs. We then attempted to rescue Oct3/4 expression in the ES-derived cells in the chimeric embryos by introducing an additional Oct3/4 transgene. The ES cell derived cells indeed recovered Oct3/4 transcription in these chimeric embryos, and were successfully specified to PGCs. We further confirmed the requirement of Oct3/4 by using another derivative of ZHBTc4 ES cells in which a Dex (dexamethasone)-dependent Oct3/4 transgene was introduced. In the presence of Dox, Oct3/4 protein was absent in the nuclei of the ES-derived cells, which failed to form PGCs. In contrast, the ES-derived cells could be specified to PGCs after activation of Oct3/4 function in the presence of Dex. PMID- 18395707 TI - Aurora-A kinase Ser349 phosphorylation is required during Xenopus laevis oocyte maturation. AB - Xenopus laevis Aurora-A is phosphorylated in vivo onto three amino acids: Ser53, Thr295 and Ser349. The activation of the kinase depends on its autophosphorylation on Thr295 within the T-loop. The phosphorylation of Ser53 by still unknown kinase(s) prevents its degradation. The present work focused on the regulation of Aurora-A function via Ser349 phosphorylation. Mutagenesis of Ser349 to alanine (S349A) had few impact in vitro on the capability of the kinase to autophosphorylate as well as on its activity. These data in addition to in gel kinase assays and site-specific proteolytic digestion experiments prove that Ser349 is clearly neither a primary autophosphorylation site, nor an autophosphorylation site depending on the priming phosphorylation of Thr295. Using specific antibodies, we also show that the phosphorylation of Aurora-A Ser349 is a physiological event during Xenopus oocyte maturation triggered by progesterone. A peak of phosphorylation paralleled the decrease of Aurora activity observed between meiosis I and II. In response to progesterone, X. laevis stage VI oocytes microinjected with the Aurora-A S349A mutant proceeded normally to germinal vesicle breakdown (GVBD), but degenerated rapidly soon after. Since phosphorylation of Ser349 is responsible for a decrease in kinase activity, our results suggest that a down-regulation of Aurora-A activity involving Ser349 phosphorylation is required in the process of maturation. PMID- 18395708 TI - The C. elegans glycosyltransferase BUS-8 has two distinct and essential roles in epidermal morphogenesis. AB - Ventral enclosure in Caenorhabditis elegans involves migration of epidermal cells over a neuroblast substrate and subsequent adhesion at the ventral midline. Organisation of the neuroblast layer by ephrins and their receptors is essential for this migration. We show that bus-8, which encodes a predicted glycosyltransferase, is essential for embryonic enclosure and acts in or with ephrin signalling to mediate neuroblast organisation and to permit epidermal migration. BUS-8 acts non-cell-autonomously in this process, and likely modifies an extracellular regulator of ephrin signalling and cell organisation. Weak and cold-sensitive alleles of bus-8 show that the gene has a separate and distinct post-embryonic role, being essential for epidermal integrity and production of the cuticle surface. This disorganisation of the epidermis and cuticle layers causes increased drug sensitivity, which could aid the growing use of C. elegans in drug screening and chemical genomics. The viable mutants are also resistant to infection by the pathogen Microbacterium nematophilum, due to failure of the bacterium to bind to the host surface. The two separate essential roles of BUS-8 in epidermal morphogenesis add to our growing understanding of the widespread importance of glycobiology in development. PMID- 18395709 TI - Metalloporphyrins as a therapeutic drug class against peroxynitrite in cardiovascular diseases involving ischemic reperfusion injury. AB - Peroxynitrite is well-recognized as being capable of inducing damaging cellular effects and has been identified as a mediator of cell damage in numerous disease states, including cardiovascular diseases. Metalloporphyrins are a class of molecule that represents an exciting new pharmacological approach to reducing peroxynitrite levels. These compounds catalyze the conversion of the harmful peroxynitrite molecules into less toxic derivatives and can be considered the reasonable intervention to reduce the toxicity of peroxynitrite. Several compounds have been synthesized and tested with promising results. Differences in the metalloporphyrin structure affect their reactivity. Iron-based metalloporphyrins display the highest rate of peroxynitrite decomposition with the narrowest scope of side reactions, whereas manganese-based metalloporphyrins react slower and with more pronounced secondary reactions, notably functioning as superoxide dismutase mimetics. This review examines the evidence that peroxynitrite is operative in patients with cardiovascular disease focusing on metalloporphyrin peroxynitrite decomposition catalysts and the evidence for their utility in the pharmacological treatment of major cardiovascular diseases. The data suggest that modification of peroxynitrite-induced cardiovascular injury is an intriguing and useful treatment approach. PMID- 18395710 TI - Neurobiology of the metabolic syndrome: an allostatic perspective. AB - The metabolic syndrome is a cluster of more or less related metabolic and cardiovascular derangements including visceral obesity, insulin resistance, blood and tissue dislipidemia, high blood pressure and it is often associated with neuroendocrine and immunological dysregulations. The aetiology of this syndrome is clinically highly relevant because it predisposes to life-threatening complications, such as Diabetes Mellitus, kidney failure, cardiovascular disease, and certain types of cancer. Contributing factors include a sedentary life-style combined with increased dietary fat intake and psychosocial stress. From a biological viewpoint, however, the metabolic syndrome can be considered as a maladaptive consequence of an initially successful adaptation to high environmental demands. As opposed to pre-historic times - when environmental demands were usually energy-costly (e.g., fight/flight/hunt) and nutritional resource often inadequate - energy-utilizing actions serve no longer an optimal solution to deal with environmental demands of current human society. This paper describes the interactions between psychosocial stress and nutrition and how these may affect emotional and metabolic components of the metabolic syndrome. A deeper understanding of these interactions is necessary to come to effective treatment and prevention of the metabolic syndrome in the future. PMID- 18395711 TI - Regulation of Akt mRNA and protein levels by glycogen synthase kinase-3beta in adrenal chromaffin cells: effects of LiCl and SB216763. AB - In cultured bovine adrenal chromaffin cells, where Akt1 is the predominant isoform over Akt2 and Akt3, chronic (> or =12 h) treatment with 1-20 mM LiCl, an inhibitor of glycogen synthase kinase-3, decreased Akt1 level by approximately 52% (EC50=3.7 mM; t1/2=l2 h); it was associated with LiCl-induced increased levels of Ser9-phosphorylated glycogen synthase kinase-3beta (approximately 37%) and beta-catenin (approximately 59%), two hallmarks of glycogen synthase kinase 3beta inhibition. The same LiCl treatment did not change phosphoinositide 3 kinase, phosphoinositide-dependent kinase 1, and extracellular signal-regulated kinase-1/2 levels. Treatment with SB216763 [3-(2,4-dichlorophenyl)-4-(1-methyl-1H indol-3-yl)-1H-pyrrole-2,5-dione], a selective inhibitor of glycogen synthase kinase-3, lowered Akt1 level by approximately 67% (EC50=2 microM; t1/2=l2 h), when SB216763 caused concentration- and time-dependent increase of beta-catenin level by approximately 76%. LiCl- or SB216763-induced Akt1 decrease, as well as increases of Ser9-phosphorylated glycogen synthase kinase-3beta and beta-catenin were restored to the control levels of nontreated cells after the washout of LiCl (20 mM for 24 h)- or SB216763 (30 microM for 24 h)-treated cells. LiCl-induced Akt1 reduction was not prevented by beta-lactone, lactacystin (two inhibitors of proteasome), calpastatin (an inhibitor of calpain), or leupeptin (an inhibitor of lysosome). LiCl decreased Akt1 mRNA level by 20% at 6 h, with no effect on Akt1 mRNA stability. These results suggest that glycogen synthase kinase-3beta inhibition caused down-regulation of Akt1 mRNA and Akt1 protein levels; conversely, constitutive activity of glycogen synthase kinase-3beta maintains steady-state level of Akt1 in quiescent adrenal chromaffin cells. PMID- 18395712 TI - Depletion of serotonin decreases the effects of the kappa-opioid receptor agonist U-69593 on cocaine-stimulated activity. AB - Treatment with a kappa-opioid receptor agonist for 5 days decreases locomotor activity and reduces activity in response to a cocaine challenge 3 days later. In addition, chronic cocaine increases kappa-opioid receptor density, striatal dynorphin, and dynorphin gene expression in the striatum. The upregulation of kappa-opioid receptors after cocaine treatment occurs predominantly in brain regions that are highly innervated by serotonin. To determine if serotonin plays a role in the effects of kappa-opioid receptor agonists on cocaine-stimulated activity, parachloroamphetamine (PCA), which depleted serotonin by 53%-66%, or saline, was given prior to a five-day treatment with U-69593 or vehicle. Three days later each rat received a single injection of cocaine and locomotor activity was measured. Treatment with PCA had no effect on the ability of U-69593 alone to decrease locomotor activity. Thus, the behavioral effects of U-69593 alone were not dependent upon serotonin. In rats pretreated with saline, U-69593 treatment significantly blocked the locomotor-activating effects of cocaine. Following PCA pretreatment, however, there were no significant differences in locomotor activity in rats challenged with an injection of cocaine after treatment with U 69593 or vehicle. Thus, serotonin depletion prevented the long-lasting blockade of the locomotor-activating effects of cocaine subsequent to repeated administration of U-69593 but did not alter the effects of cocaine in rats that were treated with vehicle. Thus, the effects of PCA on U-69593 are not due to non specific alterations in cocaine-induced locomotor activity. These findings suggest that serotonin plays an important role in mediating the effects of kappa opioid receptor agonists on the behavioral response to cocaine. PMID- 18395713 TI - Human mesenchymal stem cell proliferation is regulated by PGE2 through differential activation of cAMP-dependent protein kinase isoforms. AB - The conditions used for in vitro differentiation of hMSCs contain substances that affect the activity and expression of cyclooxygenase enzymes (COX1/COX2) and thereby the synthesis of prostanoids. hMSC constitutively produce PGE2 when cultivated in vitro. In this study we have investigated effects of PGE2 on proliferation of hMSC. We here demonstrate that one of the main control molecules in the Wnt pathway, GSK-3 beta, is phosphorylated at the negative regulatory site ser-9 after treating the cells with PGE2. This phosphorylation is mediated by elevation of cAMP and subsequent activation of PKA. Furthermore, PGE2 treatment leads to enhanced nuclear translocation of beta-catenin, thus influencing cell proliferation. The presence of two PKA isoforms, types I and II, prompted us to investigate their individual contribution in PGE2-mediated regulation of proliferation. Specific activation of PKA type II with synthetic cAMP analogues, resulted in enhancement of proliferation. On the other side, we found that treatment of hMSC with high concentrations of PGE2 inhibited cell proliferation by arresting the cells in G0/G1 phase, an effect we found to be mediated by PKA I. Hence, the two different PKA isoforms seem to have opposing functions in the regulation of proliferation and differentiation in these cells. PMID- 18395714 TI - Corneal response to femtosecond laser photodisruption in the rabbit. AB - In this report we evaluated the effect of femtosecond laser energy on the development of corneal haze and keratocyte activation in rabbits following intra stromal photodisruption to create LASIK flaps using a modified commercial femtosecond surgical laser. Three groups of flap parameters were studied: 1.5 microJ/pulse with 10 microm spot separation and complete side cut (Group 1); 3.5 microJ/pulse with 14 microm spot separation and complete side cut (Group 2); 3.5 microJ/pulse with 14 microm spot separation and partial (50 microm) side cut (Group 3). All flaps were left attached without lifting to avoid epithelial contamination. Rabbits were then evaluated pre- and post-operatively by quantitative in vivo and ex vivo confocal microscopy. The achieved flap thickness 1 week after surgery averaged 88.9+/-12.8, 90.8+/-6.9 and 86.5+/-6.8 microm for Groups 1-3 respectively (p=NS). Interface thickness was significantly greater (p<0.05) in the higher energy groups averaging 40.0+/-11.2 and 37.7+/-5.7 microm for Groups 2-3 compared to 28.6+/-4.5 microm for Group 1. Corneal haze was barely detectible and not significantly different between groups, although haze was detected in the region of the side-cuts in Groups 1 and 2. No clinically significant changes in stromal or epithelial thickness were noted. Laser confocal microscopy showed the presence of small diameter cells within the flap interface that resided within disrupted regions of the corneal collagen lamellae. Keratocyte activation was only detected in regions of the 100% side cut and not over the flap interface. In conclusion, the results of this study indicate that photodisruption of the corneal stroma alone without flap elevation regardless of laser energy does not induce significant corneal haze in the rabbit. However, a thicker stromal interface was seen with the higher energy suggesting greater stromal damage. PMID- 18395715 TI - Effects of rTMS on grip force control following subcortical stroke. AB - Within the concept of interhemispheric competition we tested the effect of inhibitory 1 Hz repetitive transcranial magnetic stimulation (rTMS), applied over the primary motor cortex of the unaffected hemisphere, upon dexterity of the affected hand in subcortical stroke patients. Subjects grasped, lifted and held an instrumented object between the index finger and thumb with both the affected and unaffected hand prior to (baseline) and following 1 Hz rTMS applied over (i) the vertex (control stimulation) and (ii) the primary motor cortex of the unaffected hemisphere. Compared to baseline, 1 Hz rTMS applied over the unaffected primary motor cortex, but not the vertex, improved the efficiency and timing of grasping and lifting with the affected hand. Our data support the interhemispheric competition concept and furthermore reinforce current efforts to implement rTMS in novel approaches to stroke rehabilitation. PMID- 18395716 TI - Coenzyme Q10 treatment in infertile men with idiopathic asthenozoospermia: a placebo-controlled, double-blind randomized trial. AB - OBJECTIVE: To evaluate the effectiveness of coenzyme Q(10) treatment in improving semen quality in men with idiopathic infertility. DESIGN: Placebo-controlled, double-blind randomized trial. SETTING: Andrology Unit, Department of Internal Medicine, Polytechnic University of Marche, Italy. PATIENT(S): Sixty infertile patients (27-39 years of age) with the following baseline sperm selection criteria: concentration >20 x 10(6)/mL, sperm forward motility <50%, and normal sperm morphology >30%; 55 patients completed the study. INTERVENTION(S): Patients underwent double-blind therapy with coenzyme Q(10), 200 mg/day, or placebo; the study design was 1 month of run-in, 6 months of therapy or placebo, and 3 months of follow-up. MAIN OUTCOME MEASURE(S): Variations in semen parameters used for patient selection and variations of coenzyme Q(10) and ubiquinol concentrations in seminal plasma and spermatozoa. RESULT(S): Coenzyme Q(10) and ubiquinol increased significantly in both seminal plasma and sperm cells after treatment, as well as spermatozoa motility. A weak linear dependence among the relative variations, baseline and after treatment, of seminal plasma or intracellular coenzyme Q(10) and ubiquinol levels and kinetic parameters was found in the treated group. Patients with a lower baseline value of motility and levels of coenzyme Q(10) had a statistically significant higher probability to be responders to the treatment. CONCLUSION(S): The exogenous administration of coenzyme Q(10) increases the level of the same and ubiquinol in semen and is effective in improving sperm kinetic features in patients affected by idiopathic asthenozoospermia. PMID- 18395717 TI - The impact of hemochromatosis mutations and transferrin genotype on gonadotropin serum levels in infertile men. AB - OBJECTIVE: To address the possibility that HFE mutations and TF gene polymorphism cause dysfunction of spermatogenesis and/or the hypothalamic-pituitary-gonadal axis via contribution to long-term iron overload in the testes and brain. DESIGN: Case-control and association study. SETTING: Clinic of obstetrics and gynecology and university-based research laboratory. PATIENT(S): 127 infertile men (including 97 with idiopathic infertility) and 188 controls of proven fertility. INTERVENTION(S): Polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP). MAIN OUTCOME MEASURE(S): HFE mutations and transferrin allelic polymorphism, and testosterone, prolactin, and gonadotropin serum levels. RESULT(S): The frequencies of the analyzed alleles and genotypes showed no statistically significant difference between infertile men and controls. Sperm count and progressive sperm motility did not correlate with HFE or TF genotype, or their combination. After excluding patients with clinical hypogonadism or varicocele from further analysis, a statistically significant correlation between serum follicle-stimulating hormone and luteinizing hormone levels and the combined HFE H63D/TFC2 genotype was found in 97 men with idiopathic infertility. CONCLUSION(S): The combined HFE H63D/TF-C2 genotype contributed to 4.1% and 10.6% of follicle-stimulating hormone and luteinizing hormone variation, respectively, in infertile men, raising mean hormonal values above the normal physiologic range. Therefore, HFE and TF genes together may influence the hypothalamic pituitary-gonadal axis, functioning at the pituitary or testes level. PMID- 18395718 TI - Impact of day 3 or day 5 embryo transfer on pregnancy rates and multiple gestations. AB - OBJECTIVE: To test the hypothesis that day 5 ET (D5ET) is superior to day 3 ET (D3ET) in pregnancy outcome and that it also reduces multiple gestations. DESIGN: Retrospective cohort study. SETTING: Assisted reproductive technologies program at Wilford Hall Medical Center. PATIENT(S): Patients electing for either D3ET or D5ET. INTERVENTION(S): Participants meeting inclusion criteria for D5ET elected either D3ET or D5ET. MAIN OUTCOME MEASURE(S): Cycles were compared by day of transfer and further stratified by patient age (<35 years and 35-40 years). The number of oocytes retrieved, embryos on day 3, embryos transferred, pregnancy rate, implantation rate, and twin and high order multiples (>or=triplets) rates were compared. RESULT(S): Of the 274 patients who met our inclusion criteria, 153 underwent a D3ET and 121 underwent a D5ET. The D5ET group had a significantly lower mean age and number of embryos transferred and a higher implantation rate (56% vs. 42%) than the D3ET group. Patients who were 35-40 years old had a significantly higher live-birth rate (68% vs. 40%). Although not statistically significant, the D5ET groups had higher clinical pregnancy (73% vs. 65%) and twin pregnancy (33% vs. 25%) rates. CONCLUSION(S): Blastocyst transfer resulted in fewer embryos transferred, with a trend toward improved clinical pregnancy and higher twin pregnancy rates. Live-birth rates were improved in patients 35-40 years of age. Younger patients opting for D5ET should do so with a commitment toward single ET. PMID- 18395719 TI - A spur sign in the EUS evaluation of dysphagia. PMID- 18395720 TI - Regional differences in the pituitary distribution of luteinizing hormone in the gonadectomized and proestrous female rat. AB - Previous data have shown that regional differences in the presence of anterior pituitary luteinizing hormone (LH) generally correlate with the comparable disparities in distribution of gonadotropes throughout the gland. In female rats, the differences are apparent over the estrous cycle, but are more prominent during the hours preceding the proestrus surge of LH. The current experiments examined (1) if such regional disparities are present throughout the surge window, (2) if differences are mirrored by release of LH in vitro and (3) if the appearance of regional differences is altered in ovariectomized females. Results showed that a comparative elevation in the rostral portion of the pituitary during the pre-surge period diminishes and finally disappears concurrent with the rise in circulating LH. This increase in rostral LH concentrations is reflected in this region by a comparable effect in vitro on stimulated LH secretion from pituitary fragments, although the effect is somewhat diminished by referencing release against tissue concentrations of LH present in a contralateral rostral fragment. Ovariectomies conducted at 1500h on proestrus, at a time when a significant regional difference has faded, resulted in a prompt increase in LH across all areas of the pituitary, and the emergence of a marked augmentation in rostral concentrations over the ensuing 72h. The effect was not seen when ovariectomies were performed on estrus. These data show that, while a regional disparity in anterior pituitary LH is present as circulating concentrations of estradiol rise prior to the LH surge, the removal of this steroid feedback at a time when LH synthesis is normally amplified accentuates the difference between the rostral region and other areas of the pituitary. PMID- 18395721 TI - John J. Sciarra IJGO Prize Paper Award. PMID- 18395722 TI - Factors associated with regular cervical cancer screening. AB - OBJECTIVE: To identify the factors associated with regular cervical screening (CS) in the French female population. METHODS: Face-to-face interviews with 5354 randomly selected French women were carried out in this population-based, observational, cross-sectional study. The variables found significant by univariate analysis were entered in a stepwise analysis. A multivariate logistic model constructed with these variables permitted to identify which were significantly related to women undergoing regular CS. RESULTS: A significant relationship was found between regular CS and previous screening for HIV infection, being professionally active, fear of cervical cancer, level of education, and having had a gynecologic check-up in the past 5 years. CONCLUSION: Since the study sample was known to be representative of the female adult population, most of our observations may have important public health implications in France, where information on HPV infection and its possible consequences need to be improved. PMID- 18395723 TI - Effect of intestinal production of equol on menopausal symptoms in women treated with soy isoflavones. AB - OBJECTIVE: To evaluate the effect of soy isoflavones on menopausal symptoms in women who do and who do not produce equol, a daidzein metabolite. METHOD: A randomized, double-blind, placebo-controlled clinical trial was conducted over 6 months with 96 healthy menopausal women. After taking take 135 mg of isoflavones daily for 1 week, the women in the study group were assigned to the equol producing (EP) or the non-EP group according to the presence or absence of equol in their urine. Menopausal symptoms were evaluated using a modified Kupperman Index. RESULT: Compared with the placebo group, the scores for hot flashes and excessive sweating were significantly reduced after 3 months, and the scores for weakness, palpitations, limb paresthesia, and total symptoms after 6 months, in the EP group only. CONCLUSIONS: Isoflavone supplementation improves menopausal symptoms only in women with the ability to produce equol. PMID- 18395724 TI - Social costs of skilled attendance at birth in rural Ghana. AB - OBJECTIVE: To examine the social costs to women of skilled attendance at birth in rural Ghana. METHOD: Ethnographic data were obtained through participant observation, interviews, case histories, and focus groups and were analyzed alongside data from a birth cohort of 2878 singletons born in the Kintampo study district between July 2003 and June 2004. RESULTS: Most women delivered at home. Home delivery raises a woman's status in her community, while seeking skilled attendance lowers it. Women feel that seeking assistance in childbirth wastes other people's time and they value secrecy in labor. Negative treatment by health providers and expensive supplies needed for delivery also act as barriers. CONCLUSION: The social costs of obtaining skilled attendance at birth must be offset by community level strategies such as mobilization of older women and husbands, and ensuring health providers extend professional, humane care to laboring women. PMID- 18395725 TI - Pelvic denervation procedures: a current reappraisal. AB - The introduction of minimally invasive techniques over recent years has led to the resurgence of pelvic denervation procedures such as presacral neurectomy and uterine nerve ablation being performed for women with dysmenorrhea and pelvic pain. Women who have failed medical therapy with persistent and debilitating symptoms may certainly benefit from these procedures. However, presacral neurectomy and uterine nerve ablation are distinct procedures that require appropriate patient selection in order to optimize pain relief. Whereas presacral neurectomy may be effective for both primary dysmenorrhea and endometriosis related pelvic pain, the role of uterine nerve ablation should be reserved for patients with primary dysmenorrhea only, as evidenced by several randomized trials. PMID- 18395726 TI - An interrater reliability study of the Braden scale in two nursing homes. AB - BACKGROUND: Adequate risk assessment is essential in pressure ulcer prevention. Assessment scales were designed to support practitioners in identifying persons at pressure ulcer risk. The Braden scale is one of the most extensively studied risk assessment instruments, although the majority of studies focused on validity rather than reliability. OBJECTIVES: The first aim was to measure the interrater reliability of the Braden scale and its individual items. The second aim was to study different statistical approaches regarding interrater reliability estimation. DESIGN AND METHODS: An interrater reliability study was conducted in two German nursing homes. Residents (n = 152) from 8 units were assessed twice. The raters were trained nurses with a work experience ranging from 0.5 to 30 years. Data were analysed using an overall percentage of agreement, weighted and unweighted kappa and the intraclass correlation coefficient. RESULTS: Differences between nurses rating the overall Braden score ranged from 0 up to 9 points. Interrater reliability expressed by the intraclass correlation coefficient ranged from 0.73 (95% CI 0.26 - 0.91) to 0.95 (95% CI 0.87 - 0.98). Calculated intraclass correlation coefficients for individual items ranged from 0.06 (95% CI -0.31 to 0.48) to 0.97 (95% CI 0.93-0.99) with the lowest values being measured for the items "sensory perception" and "nutrition". There was no association between work experience and the level of interrater reliability. With two exceptions, simple kappa-values were always lower than weighted kappa-values and intraclass correlation coefficients. CONCLUSIONS: Although the calculated interrater reliability coefficients for the total Braden score were high in some cases, several clinically relevant differences occurred between the nurses. Due to interrater reliability being very low for the items "sensory perception" and "nutrition", it is doubtful if their assessment contributes to any valid results. The calculation of weighted kappa or intraclass correlation coefficients is the most appropriate interrater reliability estimates. PMID- 18395727 TI - Older people and adherence with medication: a review of the literature. AB - BACKGROUND: Older people represent a sizeable population of the UK. Many older people receive drug treatment for long-term conditions. Adherence with medication is therefore an important clinical, financial and resource intensive concern. OBJECTIVES: This review aimed to examine patient's beliefs, perceptions and views in relation to adherence with medication. DESIGN: A comprehensive search of the literature was undertaken using numerous approaches. The search of revealed 30 research papers. FINDINGS: Articles were initially evaluated using Critical Appraisal Skills Programme principles to identify those relevant to the review. Relevant studies were then subjected to a narrative analysis to assist the development of relevant themes. Four themes were identified; experience of adherence; perceptions and attitudes to medication adherence and non-adherence; patients acceptance of their illness and impact on medication taking behaviour and shared decision making. CONCLUSIONS: The findings of this review imply that there is a need for more emphasis on shared decision making between the older patient and the prescriber. Using this approach, adherence with medication may improve. There is also a need to develop a standardized measure of medication adherence. PMID- 18395728 TI - Reduction of charge-modified LDL by statin therapy in patients with CHD or CHD risk factors and elevated LDL-C levels: the SPECIAL Study. AB - Various forms of atherogenic modified low-density lipoprotein (LDL) including oxidized LDL and small, dense LDL have increased negative charge as compared to normal LDL. Charge-modified LDL (electronegative LDL) and normal LDL subfractions in plasma are analyzed by capillary isotachophoresis (cITP) as fast-migrating LDL (fLDL) and slow-migrating LDL (sLDL). We examined the effects of pravastatin and simvastatin on charge-based LDL subfractions as determined by cITP in patients with hypercholesterolemia. Patients (n=72) with CHD or CHD risk factors and elevated LDL cholesterol (LDL-C) levels were randomly assigned to receive pravastatin or simvastatin. After treatment with statins for 3 and 6 months, both cITP fLDL and sLDL were reduced (p<0.05) from the baseline, but the effects did not differ between treatment with pravastatin and simvastatin. At baseline and after treatment for 3 months, cITP sLDL was correlated with LDL-C, but fLDL was correlated with inflammatory markers, high-sensitive C-reactive protein and LDL associated platelet-activating factor acetylhydrolase, and atherogenic lipoproteins, remnant-like particle cholesterol and small, dense LDL cholesterol. In conclusion, cITP fLDL was related to inflammatory markers and atherogenic lipoproteins and was reduced by treatment with statins. Charge-modified LDL subfraction could be a potential marker for atherosclerosis and a target for therapy. PMID- 18395729 TI - Determination of molindone enantiomers in human plasma by high-performance liquid chromatography-tandem mass spectrometry using macrocyclic antibiotic chiral stationary phases. AB - A sensitive and selective bioanalytical assay was developed and validated for the determination of enantiomeric molindone in human plasma using high-performance liquid chromatography-tandem mass spectrometry along with supported liquid extraction procedures. The chiral separation was evaluated and optimized on macrocyclic antibiotic type chiral stationary phases (CSPs) based on teicoplanin aglycone (Chirobiotic TAG) in polar organic, polar ionic, and reversed-phase mode chromatography, respectively. Complete baseline separation was achieved on a Chirobiotic TAG column under isocratic condition in reversed-phase chromatography. The method validation was conducted using a Chirobiotic TAG column (100 mm x 2.1 mm) over the curve range 0.100-100 ng/ml for each molindone enantiomer using 0.0500 ml of plasma sample. The flow rate was 0.8 ml/min and the total run time was 9 min. Supported liquid extraction in a 96-well plate format was used for sample preparation. Parameters including recovery, matrix effect, linearity, sensitivity, specificity, carryover, precision, accuracy, dilution integrity, and stability were evaluated. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels were RSD meso-tetracyclopentylcalix[4]pyrrole (TPCP)>meso-tetracyclohexylcalix[4]pyrrole (THCP). The ion-pairing constants follow the order under given solvent: Me4N+>Et4N+>Bu4N+, and ion-pairing constant of C4P follows the order: THCP>TPCP>OMCP. As though, ion-pairing association is intensified by steric hindrance. PMID- 18395732 TI - Simultaneous analysis of 16 sulfonamide and trimethoprim antibiotics in environmental waters by liquid chromatography-electrospray tandem mass spectrometry. AB - A sensitive liquid chromatography-electrospray tandem mass spectrometry method combined with solid-phase extraction and silica cartridge cleanup was established for 16 sulfonamides and trimethoprim in various water matrices. Signal suppression of all target analytes in sewage treatment plant influent, effluent and river water was improved by this method developed in this study. The method detection limits for 17 analytes were 20-200 pg/L for influent, 16-120 pg/L for effluent and 8.0-60 pg/L for river water with overall mean recoveries of 62-102% in all studied matrices. This method was used to analyze residual sulfonamides and trimethoprim in wastewater and river samples from Japan, and 8 analytes (0.08 (sulfadimethoxine)-161 ng/L (sulfapyridine) in wastewater and 10 (0.03 (sulfamethizol)-8.9 ng/L (sulfaquinoxaline) in river samples were detected. PMID- 18395733 TI - Direct synthesis of hierarchical monolithic silica for high performance liquid chromatography. AB - The silica-based monolith exhibiting a hierarchical bimodal porous structure has been directly synthesized via lytropic mesophase. The hydrolysis and condensation of tetramethoxysilane (TMOS) in the presence of poly(ethylene oxide)-block poly(propylene oxide)-block-poly(ethylene oxide) (P123) and acetic acid results in silica monolith with MSU-type mesoporous structure embedded in the skeleton of the interconnected macropore. The silica monolith with bimodal porous structure can separate benzene and phenol with high flow rate and low back-pressure. Moreover, the chromatographic property of C18-grafted silica monolith is investigated in the separation of aromatic molecules. Our primary result shows that the silica monolith with interconnected macropore and MSU-type mesopore is a promising packing material as stationary phase for high performance liquid chromatography. PMID- 18395734 TI - Micro liquid chromatography coupled with evaporative light scattering detector at ambient and high temperature: optimization of the nebulization cell geometry. AB - The recent developments in liquid chromatography (LC) are mainly dedicated to both system miniaturization (micro-, capillary-, and nano-LC) and analysis time decrease (fast-, and ultra-fast-LC). For the latter, several strategies can be used, and high temperature liquid chromatography (HTLC) seems very promising and easy to implement, especially in miniaturized system. In LC, the evaporative light scattering detector (ELSD) is considered an attractive alternative to conventional detector such as UV-vis due to its versatility and quasi universality. Therefore, the compatibility of ELSD with micro-LC and micro-HTLC was investigated for several pharmaceutical compounds of interest. The nebulization process appeared to be the most critical parameter for performing the coupling and maintaining an efficient separation. Therefore, appropriate modifications in the nebulization cell geometry were brought to make ELSD fully compatible with micro-LC. The impact of optimized nebulization cell on chromatographic performance was evaluated in terms of efficiency and sensitivity. Finally, highly efficient, sensitive and fast separations of pharmaceutical drugs were performed with both techniques and the customized nebulization cell design. PMID- 18395735 TI - Novel approach to microwave-assisted extraction and micro-solid-phase extraction from soil using graphite fibers as sorbent. AB - A single-step extraction-cleanup procedure involving microwave-assisted extraction (MAE) and micro-solid-phase extraction (micro-SPE) has been developed for the analysis of polycyclic aromatic hydrocarbons (PAHs) from soil samples. Micro-SPE is a relatively new extraction procedure that makes use of a sorbent enclosed within a sealed polypropylene membrane envelope. In the present work, for the first time, graphite fiber was used as a sorbent material for extraction. MAE-micro-SPE was used to cleanup sediment samples and to extract and preconcentrate five PAHs in sediment samples prepared as slurries with addition of water. The best extraction conditions comprised of microwave heating at 50 degrees C for a duration of 20 min, and an elution (desorption) time of 5 min using acetonitrile with sonication. Using gas chromatography (GC)-flame ionization detection (FID), the limits of detection (LODs) of the PAHs ranged between 2.2 and 3.6 ng/g. With GC-mass spectrometry (MS), LODs were between 0.0017 and 0.0057 ng/g. The linear ranges were between 0.1 and 50 or 100 microg/g for GC-FID analysis, and 1 and 500 or 1000 ng/g for GC-MS analysis. Granular activated carbon was also used for the micro-SPE device but was found to be not as efficient in the PAH extraction. The MAE-micro-SPE method was successfully used for the extraction of PAHs in river and marine sediments, demonstrating its applicability to real environmental solid matrixes. PMID- 18395736 TI - Combination of artificial neural network technique and linear free energy relationship parameters in the prediction of gradient retention times in liquid chromatography. AB - In this work multiple linear regression (MLR) and artificial neural network (ANN) were used to predict the gradient retention times of diverse sets of organic compounds in four separate data sets. Descriptors which were used as inputs of these models are five linear free energy relationship (LFER) solute parameters including E, S, A, B and V. In the first step eight separate multiple linear regression and artificial neural network models were used to predict the gradient retention time for each gradient condition separately. Results obtained in this step reveal that there are significant relations between LFER parameters and gradient retention times of solutes in liquid chromatography. Then MLR and ANN were applied to develop more general models in which several different gradient elution conditions were used. The performances of these models are compared in terms of their standard errors and also correlation analysis. The results obtained reveal that although there are no significant differences between ANN and MLR in separate modeling of the gradient retention times, ANN has a significant superiority over MLR models in developing the general models for various gradient elution conditions. The results of sensitivity analysis on ANN models indicate that the order of importance for input terms in separate ANN models is Vx>B>S>E>A and in the case of combined ANN model is Vx>B>tg>S>E>A, which are in agreement with the order of percentage of significance terms that obtained from the MLR models. PMID- 18395737 TI - Determination of oxalate in black liquor by headspace gas chromatography. AB - This study demonstrated a headspace gas chromatographic method (HS-GC) for the determination of oxalate content in black liquor (alkaline aqueous solution of inorganic chemicals and dissolved wood species from the alkaline pulping of wood). The method described in this paper is based on the reaction between oxalic and manganese dioxide in an acidic medium, in which oxalic acid is converted to carbon dioxide that is measured with a GC using a thermal conductivity detector. The challenge in developing this method was ensuring complete conversion of oxalic acid while minimizing the contribution of side reactions between carbohydrates, lignin and manganese dioxide to the carbon dioxide measured. It was found that a complete conversion of oxalate to carbon dioxide can be achieved within 3 min at a temperature of 70 degrees C; a MnO(2):C(oxalate) (concentration of H(2)C(2)O(4)+HC(2)O(4)(-)+C(2)O(4)(2-)) mole ratio of 60 and H(2)SO(4) concentration of 0.005-0.01 mol/L in the headspace vial. The method can detect concentrations as low as 0.39 microg of oxalate. The standard deviation was found to be 7% while recovery experiments with black liquor showed recoveries of 93 108% which were deemed acceptable for analysis of oxalate in an industrial sample such as black liquor. PMID- 18395738 TI - Wormlike micelles in mixed amino acid-based anionic/nonionic surfactant systems. AB - We present the formation of viscoelastic wormlike micelles in mixed amino acid based anionic and nonionic surfactants in aqueous systems in the absence of salt. N-Dodecylglutamic acid (designated as LAD) has a higher Krafft temperature; however, on neutralization with alkaline amino acid l-lysine, it forms micelles and the solution behaves like a Newtonian fluid at 25 degrees C. Addition of tri(oxyethylene) monododecyl ether (C(12)EO(3)) and tri(oxyethylene) monotetradecyl ether (C(14)EO(3)) to the dilute aqueous solution of the LAD lysine induces one-dimensional micellar growth. With increasing C(12)EO(3) or C(14)EO(3) concentration, the solution viscosity increases gradually, but after a certain concentration, the elongated micelles entangle forming a rigid network of wormlike micelles and the solution viscosity increases tremendously. Thus formed wormlike micelles show a viscoelastic character and follow the Maxwell model. Tri(oxyethylene) monohexadecyl ether (C(16)EO(3)), on the other hand, could not form wormlike micelles, although the solution viscosity increases too. The micelles become elongated; however, they do not appear to form a rigid network of wormlike micelles in the case of C(16)EO(3). Rheological measurements have shown that zero shear viscosity (eta(0)) increases with the C(12)EO(3) concentration gradually at first and then sharply, and finally decreases before phase separation. However, no such maximum in the eta(0) plot is observed with the C(14)EO(3). The eta(0) increases monotonously with the C(14)EO(3) concentration till phase separation. In studies of the effect of temperature on the wormlike micellar behavior it has been found that the eta(0) decays exponentially with temperature, following an Arrehenius behavior and at sufficiently higher temperatures the solutions follow a Newtonian behavior. The flow activation energy calculated from the slope of log eta(0) versus 1/T plot is very close to the value reported for typical wormlike micelles. Finally, we also present the effect of neutralization degree of lysine on the rheology and phase behavior. The formation of wormlike micelles is confirmed by the Maxwell model fit to the experimental rheological data and by Cole-Cole plots. PMID- 18395739 TI - Synthesis of poly(N-isopropylacrylamide)-b-poly(2-vinylpyridine) block copolymers via RAFT polymerization and micellization behavior in aqueous solution. AB - Poly(N-isopropylacrylamide)-b-poly(2-vinylpyridine) (PNIPAM-b-P2VP) block copolymers were synthesized for the first time via reversible addition fragmentation chain transfer (RAFT) polymerization in the presence of S-1-dodecyl S(')-(a,a(')-dimethyl-a('')-acetic acid)trithiocarbonate as chain transfer agent (CTA) and 2,2(')-azobis(isobutyronitrile) as initiator. Both pH- and thermo induced micellization behavior of the PNIPAM(59)-b-P2VP(102) block copolymer in dilute aqueous solution was investigated by pyrene fluorescence, dynamic and static light scattering, transmission electron microscopy and (1)H NMR. The results show that the critical aggregation pH value of the block copolymer is around 5 and the critical aggregation temperature of the block copolymer is around 42 degrees C. A reversible transition between P2VP-core and PNIPAM-core micelles can be observed through an intermediate unimer state in aqueous solution. PMID- 18395740 TI - Dynamics of lipid accumulation by the fat body of Rhodnius prolixus: the involvement of lipophorin binding sites. AB - In insects, lipids are stored in the fat body, mainly as triacylglycerol (TAG). In Rhodnius prolixus, a hematophagous hemipteran, lipids are accumulated after blood meal to be used later on. In adult females, at the second day after feeding, the amount of TAG was 57+/-17 microg/fat body, it increased almost five times and at fourth day it was 244+/-35 microg/fat body. TAG content remained constant until day 13, but it then decreased and, at day 20th it was very low (31+/-4.9 microg/fat body). Radiolabeled free fatty acid was used to follow lipid accumulation by the fat body, as it was previously shown that, in R. prolixus, injected free fatty acids associate with lipophorin, a major hemolymphatic lipoprotein. (3)H-palmitic acid was injected into the hemocoel of R. prolixus females. It disappeared from the hemolymph very rapidly, and radioactivity was incorporated by the fat body. Sixty minutes after injection, radioactivity in the fat body was found mainly in TAGs. The capacity of the fat body to incorporate fatty acids from the hemolymph varied according to the days after blood meal, and it was maximal around the fourth day. Lipophorin binding to specific sites in fat body membrane preparations also showed variation at different days. When membranes obtained from insects at the second, fifth and tenth days were compared, binding was highest at fifth day after feeding. PMID- 18395741 TI - New cell lines derived from the black cutworm, Agrotis ipsilon, that support replication of the A. ipsilon multiple nucleopolyhedrovirus and several group I nucleopolyhedroviruses. AB - New cell lines were recently developed from the embryos of the black cutworm, Agrotis ipsilon (Lepidoptera: Noctuidae). A primary culture was initiated from 4 day-old A. ipsilon eggs in ExCell420 medium supplemented with 5% fetal bovine serum. This initial culture produced sufficient cell growth to allow subcultivation and eventually led to the establishment of eight distinct strains. Two of these strains (AiE1611T and AiEd6T) were selected for further characterization. Extracts of these strains were compared to an extract from A. ipsilon eggs by isozyme analysis and shown to be from the same species. Both strains were susceptible to infection by the A. ipsilon multiple nucleopolyhedrovirus (AgipMNPV), as well as to lepidopteran group I NPVs from A. californica, Anagrapha falcifera, Anticarsia gemmatalis, Galleria mellonella, Helicoverpa armigera, Plutella xylostella, and Rachiplusia ou, with large numbers of occlusion bodies produced in most of the inoculated cells. The cell lines did not support the replication of group II NPVs from Helicoverpa zea, Lymantria dispar, and Spodoptera exigua. Both cell lines produced confluent monolayers in plaque assays and supported the formation of plaques upon infection with AgipMNPV and Autographa californica (Ac)MNPV. Twenty AgipMNPV plaques were picked from either AiE1611T or AiEd6T monolayers, and the plaque isolates were serially passaged three times through A. ipsilon cells. Only one isolate from AiE1611T cells exhibited genotypic variation in the form of an altered restriction fragment profile. Our results suggest these new lines can be useful in the study of AgipMNPV and A. ipsilon cellular and molecular biology. PMID- 18395742 TI - Substrate recognition mechanism of alpha-1,6-glucosidic linkage hydrolyzing enzyme, dextran glucosidase from Streptococcus mutans. AB - We have determined the crystal structure of Streptococcus mutans dextran glucosidase, which hydrolyzes the alpha-1,6-glucosidic linkage of isomaltooligosaccharides from their non-reducing ends to produce alpha-glucose. By using the mutant of catalytic acid Glu236-->Gln, its complex structure with the isomaltotriose, a natural substrate of this enzyme, has been determined. The enzyme has 536 amino acid residues and a molecular mass of 62,001 Da. The native and the complex structures were determined by the molecular replacement method and refined to 2.2 A resolution, resulting in a final R-factor of 18.3% for significant reflections in the native structure and 18.4% in the complex structure. The enzyme is composed of three domains, A, B and C, and has a (beta/alpha)(8)-barrel in domain A, which is common to the alpha-amylase family enzymes. Three catalytic residues are located at the bottom of the active site pocket and the bound isomaltotriose occupies subsites -1 to +2. The environment of the glucose residue at subsite -1 is similar to the environment of this residue in the alpha-amylase family. Hydrogen bonds between Asp60 and Arg398 and O4 atom of the glucose unit at subsite -1 accomplish recognition of the non reducing end of the bound substrate. The side-chain atoms of Glu371 and Lys275 form hydrogen bonds with the O2 and O3 atoms of the glucose residue at subsite +1. The positions of atoms that compose the scissile alpha-1,6-glucosidic linkage (C1, O6 and C6 atoms) are identical with the positions of the atoms in the scissile alpha-1,4 linkage (C1, O4 and C4 atoms) of maltopentaose in the alpha amylase structure from Bacillus subtilis. The comparison with the alpha-amylase suggests that Val195 of the dextran glucosidase and the corresponding residues of alpha-1,6-hydrolyzing enzymes participate in the determination of the substrate specificity of these enzymes. PMID- 18395744 TI - Crystal structure of the peptidoglycan recognition protein at 1.8 A resolution reveals dual strategy to combat infection through two independent functional homodimers. AB - The mammalian peptidoglycan recognition protein-S (PGRP-S) binds to peptidoglycans (PGNs), which are essential components of the cell wall of bacteria. The protein was isolated from the samples of milk obtained from camels with mastitis and purified to homogeneity and crystallized. The crystals belong to orthorhombic space group I222 with a=87.0 A, b=101.7 A and c=162.3 A having four crystallographically independent molecules in the asymmetric unit. The structure has been determined using X-ray crystallographic data and refined to 1.8 A resolution. Overall, the structures of all the four crystallographically independent molecules are identical. The folding of PGRP-S consists of a central beta-sheet with five beta-strands, four parallel and one antiparallel, and three alpha-helices. This protein fold provides two functional sites. The first of these is the PGN-binding site, located on the groove that opens on the surface in the direction opposite to the location of the N terminus. The second site is implicated to be involved in the binding of non-PGN molecules, it also includes putative N-terminal segment residues (1-31) and helix alpha2 in the extended binding. The structure reveals a novel arrangement of PGRP-S molecules in which two pairs of molecules associate to form two independent dimers. The first dimer is formed by two molecules with N-terminal segments at the interface in which non PGN binding sites are buried completely, whereas the PGN-binding sites of two participating molecules are fully exposed at the opposite ends of the dimer. In the second dimer, PGN-binding sites are buried at the interface while non-PGN binding sites are fully exposed at the opposite ends of the dimer. This form of dimeric arrangement is unique and seems to be aimed at enhancing the capability of the protein against specific invading bacteria. This mode of functional dimerization enhances efficiency and specificity, and is observed for the first time in the family of PGRP molecules. PMID- 18395743 TI - Kinetic control of Mg2+-dependent melting of duplex DNA ends by Escherichia coli RecBC. AB - Escherichia coli RecBCD is a highly processive DNA helicase involved in double strand break repair and recombination that possesses two helicase/translocase subunits with opposite translocation directionality (RecB (3' to 5') and RecD (5' to 3')). RecBCD has been shown to melt out approximately 5-6 bp upon binding to a blunt-ended duplex DNA in a Mg(2+)-dependent, but ATP-independent reaction. Here, we examine the binding of E. coli RecBC helicase (minus RecD), also a processive helicase, to duplex DNA ends in the presence and in the absence of Mg(2+) in order to determine if RecBC can also melt a duplex DNA end in the absence of ATP. Equilibrium binding of RecBC to DNA substrates with ends possessing pre-formed 3' and/or 5' single-stranded (ss)-(dT)(n) flanking regions (tails) (n ranging from zero to 20 nt) was examined by competition with a fluorescently labeled reference DNA and by isothermal titration calorimetry. The presence of Mg(2+) enhances the affinity of RecBC for DNA ends possessing 3' or 5'-(dT)(n) ssDNA tails with n<6 nt, with the relative enhancement decreasing as n increases from zero to six nt. No effect of Mg(2+) was observed for either the binding constant or the enthalpy of binding (Delta H(obs)) for RecBC binding to DNA with ssDNA tail lengths, n>or=6 nucleotides. Upon RecBC binding to a blunt duplex DNA end in the presence of Mg(2+), at least 4 bp at the duplex end become accessible to KMnO(4) attack, consistent with melting of the duplex end. Since Mg(2+) has no effect on the affinity or binding enthalpy of RecBC for a DNA end that is fully pre-melted, this suggests that the role of Mg(2+) is to overcome a kinetic barrier to melting of the DNA by RecBC and presumably also by RecBCD. These data also provide an accurate estimate (Delta H(obs)=8+/-1 kcal/mol) for the average enthalpy change associated with the melting of a DNA base-pair by RecBC. PMID- 18395745 TI - Salt-bridge dynamics control substrate-induced conformational change in the membrane transporter GlpT. AB - Active transport of substrates across cytoplasmic membranes is of great physiological, medical and pharmaceutical importance. The glycerol-3-phosphate (G3P) transporter (GlpT) of the E. coli inner membrane is a secondary active antiporter from the ubiquitous major facilitator superfamily that couples the import of G3P to the efflux of inorganic phosphate (P(i)) down its concentration gradient. Integrating information from a novel combination of structural, molecular dynamics simulations and biochemical studies, we identify the residues involved directly in binding of substrate to the inward-facing conformation of GlpT, thus defining the structural basis for the substrate-specificity of this transporter. The substrate binding mechanism involves protonation of a histidine residue at the binding site. Furthermore, our data suggest that the formation and breaking of inter- and intradomain salt bridges control the conformational change of the transporter that accompanies substrate translocation across the membrane. The mechanism we propose may be a paradigm for organophosphate:phosphate antiporters. PMID- 18395746 TI - Tet repressor induction by tetracycline: a molecular dynamics, continuum electrostatics, and crystallographic study. AB - The Tet repressor (TetR) mediates the most important mechanism of bacterial resistance against tetracycline (Tc) antibiotics. In the absence of Tc, TetR is tightly bound to its operator DNA; upon binding of Tc with an associated Mg(2+) ion, it dissociates from the DNA, allowing expression of the repressed genes. Its tight control by Tc makes TetR broadly useful in genetic engineering. The Tc binding site is over 20 A from the DNA, so the binding signal must propagate a long distance. We use molecular dynamics simulations and continuum electrostatic calculations to test two models of the allosteric mechanism. We simulate the TetR:DNA complex, the Tc-bound, "induced" TetR, and the transition pathway between them. The simulations support the model inferred previously from the crystal structures and reveal new details. When [Tc:Mg](+) binds, the Mg(2+) ion makes direct and water-mediated interactions with helix 8 of one TetR monomer and helix 6 of the other monomer, and helix 6 is pulled in towards the central core of the structure. Hydrophobic interactions with helix 6 then pull helix 4 in a pendulum motion, with a maximal displacement at its N-terminus: the DNA interface. The crystal structure of an additional TetR reported here corroborates this motion. The N-terminal residue of helix 4, Lys48, is highly conserved in DNA binding regulatory proteins of the TetR class and makes the largest contribution of any amino acid to the TetR:DNA binding free energy. Thus, the conformational changes lead to a drastic reduction in the TetR:DNA binding affinity, allowing TetR to detach itself from the DNA. Tc plays the role of a specific Mg(2+) carrier, whereas the Mg(2+) ion itself makes key interactions that trigger the allosteric transition in the TetR:Tc complex. PMID- 18395747 TI - Structural basis for autoinhibition of ESCRT-III CHMP3. AB - Endosomal sorting complexes required for transport (ESCRT-0, ESCRT-I, ESCRT-II, and ESCRT-III) are selectively recruited to cellular membranes to exert their function in diverse processes, such as multivesicular body biogenesis, enveloped virus budding, and cytokinesis. ESCRT-III is composed of members of the charged multivesicular body protein (CHMP) family--cytosolic proteins that are targeted to membranes via yet unknown signals. Membrane targeting is thought to result in a membrane-associated protein network that presumably acts at a late budding step. Here we provide structural evidence based on small-angle X-ray scattering data that ESCRT-III CHMP3 can adopt two conformations in solution: a closed globular form that most likely represents the cytosolic conformation and an open extended conformation that might represent the activated form of CHMP3. Both the closed and open conformations of CHMP3 interact with AMSH with high affinity. Although the C-terminal region of CHMP3 is required for AMSH interaction, a peptide thereof reveals only weak binding to AMSH, suggesting that other regions of CHMP3 contribute to the high-affinity interaction. Thus, AMSH, including its MIT (microtubule interacting and transport) domain, interacts with ESCRT-III CHMP3 differently from reported Vps4 MIT domain-CHMP protein interactions. PMID- 18395748 TI - Vascular prostheses: performance related to cell-shear responses. AB - BACKGROUND: This work concerned the endothelialization of vascular prostheses and subsequent improvement of functionality with respect to tissue engineering. The aim of the study was to investigate the initial, pre-shear stress cellular behavior with respect to three vascular biomaterials to explain subsequent cellular responses to physiological shear stresses. MATERIALS AND METHODS: Expanded polytetrafluoroethylene (ePTFE), polyethyleneterephthalate (polyester; Dacron; PET), and electrostatically spun polyurethane (PU) (all pre-impregnated with collagen I/III) were cell-seeded with L929 immortalized murine fibroblasts or human umbilical vein endothelial cells (HUVECs). Cytoskeletal involvement, cell height profiles, and immunohistochemistry were examined after 7 d static culture. RESULTS: All three vascular biomaterials demonstrated different structures. Cell behavior varied both between the materials and the two cell types: cytoskeletal involvement was greater for the HUVECs and the more fibrous surfaces; height profiles were greater for the L929 and PET, and lowest on PU. Immunohistochemistry of HUVEC samples also showed differences: PU revealed the greatest expression of intercellular adhesion molecule-1 and E-selectin (PET and ePTFE the lowest, respectively); ePTFE produced the greatest for vascular cell adhesion molecule-1 (PET the lowest). CONCLUSIONS: Material substrate influenced the cellular response. Cells demonstrating firm adhesion increased their cytoskeletal processes and expression of cell-substratum and inter-cellular adhesion markers, which may explain their ability to adapt more readily to shear stress. The fibrous PU structure appeared to be most suited to further shear stress exposure. This study demonstrated the potential of the underlying vascular material to affect the long-term cellular functionality of the prosthesis. PMID- 18395749 TI - EH Networks as a scaffold for skeletal muscle regeneration in abdominal wall hernia repair. AB - Incisional hernias are a common clinical problem occurring in up to 10% of all patients undergoing abdominal procedures. Primary closure, synthetic biomaterials, as well as xenografts and allografts have been used in hernia defect repair. Despite these approaches, the incidence of hernia recurrence ranges from 32% to 63%. To address this high recurrence rate, we propose an incisional hernia treatment that utilizes a functional biomaterial developed for skeletal muscle regeneration. In particular, we have developed a cyclic acetal biomaterial (EH network) based on 5-ethyl-5-(hydroxymethyl)-beta,beta-dimethyl 1,3-dioxane-2-ethanol diacrylate. Initial tests of the scaffold's mechanical properties indicate that the complex modulus of the EH network decreased after a significant increase in initiator concentration. Subsequent studies indicate that EH networks promote myoblastic cell attachment and proliferation as well as delivers functional insulin-like growth factor-1 to an in vitro population of skeletal myoblasts. This work establishes that an EH network, a degradable cyclic acetal biomaterial, can function as a scaffold for skeletal muscle engineering. PMID- 18395750 TI - Interleukin-1alpha enhances angiogenesis and is associated with liver metastatic potential in human gastric cancer cell lines. AB - BACKGROUND: To better understand the underlying mechanism of liver metastasis formation in human gastric cancer, we evaluated the angiogenic capabilities of human gastric cancer cell lines with different metastatic potentials as well as the role of interleukin (IL)-1alpha in the angiogenic process. MATERIALS AND METHODS: Reverse transcription-polymerase chain reaction was used to detect the expression of IL-1alpha and vascular endothelial growth factor (VEGF) mRNA in gastric cancer cell lines with different liver metastatic potentials. Levels of VEGF secreted by human gastric cancer cells were measured by enzyme-linked immunosorbent assay. We also examined how gastric cancer cells with different metastatic potentials influence the proliferation and tube formation of human umbilical vein endothelial cells (HUVECs) using the Premix WST-1 cell proliferation assay system and an angiogenesis assay, respectively. RESULTS: IL 1alpha expression levels were significantly correlated with liver metastatic potential in gastric cancer cell lines. Levels of VEGF secreted by gastric cancer cells appear to be regulated by IL-1alpha through IL-1 receptor Type 1 and were correlated with liver metastatic potential. Both HUVEC proliferation and tube formation were strongly enhanced by coculture with high liver-metastatic gastric cancer cells and were enhanced to a similar extent by culture in the presence of IL-1alpha. In contrast, blockade of IL-1alpha inhibited both HUVEC proliferation and angiogenesis. CONCLUSIONS: IL-1alpha may play a role in liver metastasis of gastric cancer via enhanced vascular endothelial cell proliferation and angiogenesis. PMID- 18395751 TI - Bimodal electric tissue ablation-long term studies of morbidity and pathological change. AB - BACKGROUND: Radiofrequency ablation is a popular method of treating unresectable liver tumors but tumors greater than 3 cm in diameter have a much greater risk of local recurrence after treatment. Bimodal electric tissue ablation is a modified form of radiofrequency ablation that creates significantly larger ablations by the addition of extra direct current circuitry. This may help to reduce the risk of local recurrence in these larger tumors. Prior to use in a clinical setting, a long term study was performed to assess associated morbidity and the pathological changes in the ablations. METHODS: In eight pigs, six ablations were performed in each liver. Pigs were euthanized at 2 d, 2 wk, 2 mo, and 4 mo, and the ablations were assessed macroscopically and microscopically for pathological change. Regular blood tests were performed to assess changes in liver function. At death, any other abnormalities detected were reported. RESULTS: Histopathological examination of ablation zones revealed tissue death by coagulative necrosis and healing by fibrotic scarring. Transient rises in serum liver enzymes were seen in the postoperative period. Skin necrosis was noted at the site of the positive electrode of the direct electrical current but no other form of morbidity was seen associated with the procedure. CONCLUSIONS: Although the positive electrode placement requires further consideration, bimodal electric tissue ablation appears to be safe and behaves in a similar fashion to other thermal therapies such as standard radiofrequency ablation. PMID- 18395752 TI - Pitavastatin inhibits intimal hyperplasia in rabbit vein graft. AB - BACKGROUND: The autologous saphenous vein graft is currently the most suitable conduit for arterial bypass of the lower limbs. Various approaches have been attempted to control vein graft intimal hyperplasia, and several recent reports have suggested that statin use may be linked to improved patency of vein grafts. In this study, the efficacy of pitavastatin was evaluated on intimal hyperplasia and midkine expression of experimental normocholesterolemic rabbit autologous vein graft. MATERIALS AND METHODS: Rabbits were fed regular rabbit chow, and in half of them, pitavastatin (1 mg/kg/d) was administered. A week after starting the treatment, jugular vein was implanted into the carotid artery. At 2 and 4 wk after the operation, vein grafts were harvested, and intimal hyperplasia of vein grafts were assessed. Cell proliferation in neointima was determined by proliferative cell nuclear antigen and Ki-67 stain 2 wk after implantation. In addition, the effect of pitavastatin on midkine, a heparin-binding growth factor, expressed in vein grafts was analyzed by Western blotting. RESULTS: The intimal hyperplasia in the pitavastatin group was significantly suppressed compared with the control group. Both proliferative cell nuclear antigen and Ki-67 labeling index were significantly lower in the pitavastatin group, and pitavastatin significantly reduced midkine expression of vein graft. CONCLUSIONS: These results demonstrate the efficacy of pitavastatin in reducing the degree of intimal hyperplasia of rabbit autologous vein grafts under normocholesterolemic condition. The mechanism of inhibition of intimal hyperplasia might be associated with midkine suppression. PMID- 18395753 TI - Hemoglobin-glutamer 200 reduces reperfusion injury of the cold preserved rat liver by induction of heme oxygenase-1. AB - Microcirculatory failure after cold liver preservation and reperfusion impairs tissue oxygenation and causes additional organ damage. Hemoglobin-glutamer (HbG) 200 is a hemoglobin-based oxygen carrying solution capable to improve organ oxygenation. The aim of this study was to evaluate its potential to decrease reperfusion injury after cold liver preservation. Therefore, Wistar rat livers were stored at 4 degrees C for 24 h and reperfused in the isolated perfused rat liver model with a sanguineous perfusate for 180 min. The perfusate consisted of rat blood and Krebs-Henseleit solution (Group A), supplemented by either HES 6% (Group B), or HbG (Groups C and D). In Group D heme oxygenase (HO) activity was blocked by intraperitoneal tin protoporphyrin-IX application before organ harvest. HbG supplementation increased the perfusate hemoglobin by 3,3 g/dL. After 180 min reperfusion perfusate alanine aminotransferase levels (72 +/- 27 micro/L) were significantly reduced in Group C, compared with Groups A and B (140 +/- 28 micro/L and 203 +/- 62 micro/L, respectively). These results correlated with a significant increase of HO-1 expression and activity during reperfusion. These effects could be abolished by tin protoporphyrin-IX application. HbG has been proven to be effective to reduce cold liver preservation-reperfusion injury. The positive effect on reperfusion injury depends on the induction of HO-1, which increases the bilirubin production, an important antioxidant acting as intracellular radical scavenger. PMID- 18395754 TI - Pre-hospital delay in the use of intravenous rt-PA for acute ischemic stroke in Japan. AB - In this study we investigated the factors associated with pre-hospital delay to treat acute ischemic stroke and transient attack with intravenous recombinant tissue-plasminogen activator (rt-PA) in Japan. In 625 patients, we investigated the pathways and times of their arrival to our hospital, and the significant and independent factors in the patients' clinical backgrounds associated with delayed arrival (>2 h after notice). In total, 287 patients arrived at our hospital directly via EMS, 113 came by themselves, and 225 transferred from other institutes. Delayed arrivals occurred in 423 patients (68%). Multivariate analyses showed that staying in another hospital at notice, a worsened course, and referral from other institutes were positively related, and evening onset, having a witness at onset, loss of consciousness, and using EMS were negatively related to delayed arrival; a worsened course was positively related, and staying in other hospital at notice, having a witness at onset, loss of consciousness, and a high NIHSS on admission were negatively related to delayed alert; hypercholesterolemia and onset in a nursing home were positively related, and staying in other hospital at notice, loss of consciousness, and a high NIHSS on admission were negatively related to not using EMS. A lack of knowledge concerning stroke emergency by medical staff as well as the general public may be responsible for some stroke patients losing the chance for rt-PA treatment. PMID- 18395755 TI - Spatial distribution of VEGF isoforms and chemotactic signals in the vicinity of a tumor. AB - We propose a mathematical model that describes the formation of gradients of different isoforms of vascular endothelial growth factor (VEGF). VEGF is crucial in the process of tumor-induced angiogenesis, and recent experiments strongly suggest that the molecule is most potent when bound to the extracellular matrix (ECM). Using a system of reaction-diffusion equations, we study diffusion of VEGF, binding of VEGF to the ECM, and cleavage of VEGF from the ECM by matrix metalloproteases (MMPs). We find that spontaneous gradients of matrix-bound VEGF are possible for an isoform that binds weakly to the ECM (i.e. VEGF(165)), but cleavage by MMPs is required to form long-range gradients of isoforms that bind rapidly to the ECM (i.e. VEGF(189)). We also find that gradient strengths and ranges are regulated by MMPs. Finally, we find that VEGF molecules cleaved from the ECM may be distributed in patterns that are not conducive to chemotactic migration toward a tumor, depending on the spatial distribution of MMP molecules. Our model elegantly explains a number of in vivo observations concerning the significance of different VEGF isoforms, points to VEGF(165) as an especially significant therapeutic target and indicator of a tumor's angiogenic potential, and enables predictions that are subject to testing with in vitro experiments. PMID- 18395756 TI - L-triiodothyronine differentially and nongenomically regulates synaptosomal protein phosphorylation in adult rat brain cerebral cortex: role of calcium and calmodulin. AB - Adult-onset thyroid disorders in humans impair several important central nervous system functions, causing various neuropsychiatric diseases. However, the mechanisms of thyroid hormone (TH) action in the mature mammalian brain remain unclear. Recent nongenomic actions of TH in adult brains are spotlighted. Many nongenomic mechanisms are modulated by phosphorylation-dephosphorylation of substrate proteins. In the present study, L-triiodothyronine (L-T3) demonstrated differential regulation of phosphorylation status of five different synaptosomal proteins (63, 53, 38, 23, and 16 kD) in both a Ca(2+)/calmodulin (CaM)-dependent and -independent manner. L-T3 increased the level of phosphorylation of all these five proteins. Ca(2+)/CaM further stimulated phosphorylation of 63- and 53-kD proteins by L-T3, which were inhibited both by EGTA (Ca(2+)-chelator) or KN62 (Ca(2+)/CaM kinase-II [CaMK-II] inhibitor), suggesting the role of CaMK-II. L-T3 increased the phosphorylation of 23- and 38-kD proteins; the effect was independent of EGTA or KN62. The presence of Ca(2+) decreased L-T3-induced phosphorylation of 63-, 53- and 38-kD proteins. Surprisingly, l-T3-induced phosphorylation of 16-kD protein was not augmented further with Ca(2+) or Ca(2+)/CaM; instead, the presence of CaM abolished the L-T3-induced phosphorylation. EGTA or KN62 could not restore the effect of CaM-induced dephosphorylation of this protein. This study identified the role of Ca(2+)/CaM in the regulation of L-T3-induced protein phosphorylation and supported a unique nongenomic mechanism of second messenger-mediated regulation of protein phosphorylation by TH in mature rat brain. This has profound implications for higher mental functions and strategies for novel therapeutics. PMID- 18395757 TI - Pyrolysis of spill oils adsorbed on zeolites with product oils recycling. AB - Experimentally, a feasibility study for adsorption and catalytic pyrolysis of spill oils on Cu/ZSM-5 for recycling of light oils has been conducted in the present work. The adsorption and pyrolysis of model compounds such as heptane, toluene, and diesel (to stimulate the spill oils) on Cu/ZSM-5 have been investigated on a continuous fixed-bed reactor. By component fitted X-ray absorption near edge structural (XANES) spectroscopy, catalytic active species such as metallic copper (Cu) (77-84%) and Cu(2)O (6-7%) are found in the channels of ZSM-5 during pyrolysis of heptane or toluene. Pyrolysis of diesel effected by Cu/ZSM-5 yields gas (C(1)-C(5)) (32%) and light oil (68%) that can be used as auxiliary fuels. PMID- 18395758 TI - Evaluation of induction of metallothionein-like proteins (MTLPs) in the polychaetes for biomonitoring of heavy metal pollution in marine sediments. AB - Polychaetes are suitable organisms for evaluation of impact of sediment pollution. We evaluated toxicity of cadmium and copper and measured metallothionein-like proteins (MTLPs) in the polychaete Perinereis nuntia. At the same concentration ranges copper was unexpectedly more toxic than cadmium. Copper also caused no significant increase in MTLPs in the polychaetes. When P. nuntia and another polychaete species, Cirratulus cirratus were cultured in the contaminated sediments collected from Lake Sihwa (Korea), a high mortality of 80% was observed on day 6 in P. nuntia in the sediment with the highest metal concentration. However, no mortality was observed up to 35 days in C. cirratus in any sediment. MTLP contents between two species also varied. These findings suggest that MTLP induction response in the polychaetes varies with the metal type and species and it may be used as a biomarker of sediment pollution in the polychaetes after further validation and field trials. PMID- 18395759 TI - Intracerebroventricular injection of propionic acid, an enteric bacterial metabolic end-product, impairs social behavior in the rat: implications for an animal model of autism. AB - Environmental, dietary, and gastrointestinal factors may contribute to autism spectrum disorders (ASD). Propionic acid (PPA) is a short chain fatty acid, a metabolic end-product of enteric bacteria in the gut, and a common food preservative. Recent evidence indicates that PPA can cause behavioral abnormalities and a neuroinflammatory response in rats. Social behavior was examined in similarly-treated pairs of adult male Long-Evans rats placed in an open field following intracerebroventricular (ICV) injection of PPA (4 microl of 0.26 M solution) or control compounds. Behavior was analyzed using both the EthoVision behavior tracking system and by blind scoring of videotapes of social behaviors. Compared to controls, rats treated with PPA displayed social behavior impairments as indicated by significantly greater mean distance apart, reduced time spent in close proximity, reduced playful interaction, and altered responses to playful initiations. Treatment with another short chain fatty acid, sodium acetate, produced similar impairments, but treatment with the alcohol analog of PPA, 1-propanol, did not produce impairments. Immunohistochemical analysis of brain tissue taken from rats treated with PPA revealed reactive astrogliosis, indicating a neuroinflammatory response. These findings suggest that PPA can change both brain and behavior in the laboratory rat in a manner that is consistent with symptoms of human ASD. PMID- 18395761 TI - Longitudinal studies of semantic dementia: the relationship between structural and functional changes over time. AB - The pattern of brain atrophy in semantic dementia and its associated cognitive effects have attracted a considerable body of research, but the nature of core impairments remains disputed. A key issue is whether the disease encompasses one neurocognitive network (semantics) or two (language and semantics). In order to address these conflicting perspectives, we conducted a longitudinal investigation of two semantic dementia patients, in which behavioural performance across a range of measures of language and semantic performance was assessed and interpreted in the context of annually acquired MRI scans. Our results indicated a core semantic impairment in early stages of the disease, associated with atrophy of the inferior, anterior temporal cortex. Linguistic impairments emerged later, and were contingent on atrophy having spread into areas widely believed to subserve core language processes (left posterior perisylvian, inferior frontal and insular cortex). We claim, therefore, that phonological, syntactic and morphological processing deficits in semantic dementia reflect damage to core language areas. Further, we propose that much of the current controversy over the nature of deficits in semantic dementia reflect a tendency in the literature to adopt a static perspective on what is a progressive disease. An approach in which the relationship between progressive neural changes and behavioural change over time is carefully mapped, offers a more constraining data-set from which to draw inferences about the relationship between language, semantics and the brain. PMID- 18395762 TI - Flavonols from Pterogyne nitens and their evaluation as myeloperoxidase inhibitors. AB - A myeloperoxidase inhibitory kaempferol derivative, namely pterogynoside (1), was isolated from fruits of Pterogyne nitens, along with six known flavonols, kaempferol, afzelin, kaempferitrin, quercetin, isoquercetrin and rutin. The structures of all compounds were elucidated primarily from 1D and 2D NMR spectroscopic analyses, as well as by high resolution mass spectrometry. All flavonols were screened to identify secondary metabolites as potential myeloperoxidase (MPO) inhibitors, and at concentrations of 0.50-50nM, quercetin (5), isoquercitrin (6) and rutin (7) exhibited strong inhibitory effects with IC50 values of 1.22+/-0.01, 3.75+/-0.02 and 3.60+/-0.02, respectively. The MPO activity detected for the new derivative 1 was markedly decreased (IC(50) 10.3+/ 0.03) when compared with known flavonols 5-7, and interestingly increased when tested against ABTS scavenging activity. PMID- 18395760 TI - Mapping progressive brain structural changes in early Alzheimer's disease and mild cognitive impairment. AB - Alzheimer's disease (AD), the most common neurodegenerative disorder of the elderly, ranks third in health care cost after heart disease and cancer. Given the disproportionate aging of the population in all developed countries, the socio-economic impact of AD will continue to rise. Mild cognitive impairment (MCI), a transitional state between normal aging and dementia, carries a four- to sixfold increased risk of future diagnosis of dementia. As complete drug-induced reversal of AD symptoms seems unlikely, researchers are now focusing on the earliest stages of AD where a therapeutic intervention is likely to realize the greatest impact. Recently neuroimaging has received significant scientific consideration as a promising in vivo disease-tracking modality that can also provide potential surrogate biomarkers for therapeutic trials. While several volumetric techniques laid the foundation of the neuroimaging research in AD and MCI, more precise computational anatomy techniques have recently become available. This new technology detects and visualizes discrete changes in cortical and hippocampal integrity and tracks the spread of AD pathology throughout the living brain. Related methods can visualize regionally specific correlations between brain atrophy and important proxy measures of disease such as neuropsychological tests, age of onset or factors that may influence disease progression. We describe extensively validated cortical and hippocampal mapping techniques that are sensitive to clinically relevant changes even in the single individual, and can identify group differences in epidemiological studies or clinical treatment trials. We give an overview of some recent neuroimaging advances in AD and MCI and discuss strengths and weaknesses of the various analytic approaches. PMID- 18395763 TI - Water, water, everywhere, nor any drop to drink. PMID- 18395764 TI - Non-linear density dependence in time series is not evidence of non-logistic growth. AB - Time series of population density are often used to seek deviations from logistic regulation by testing for a non-linear decline in per capita growth rate with density. Here I show that this method fails when the interval between observations is not matched by the timing of density impacts on growth. Time series overestimate instantaneous density impacts at low density and underestimate them at high density. More generally, logistic growth produces a deterministically decelerating decline in per capita growth with density if the interval between measures of population size exceeds any lag in density response. Deceleration arises independently out of stochastic density fluctuations, and under-compensating regulation. These multiple influences lead to the conclusion that sequential density estimates provide insufficient information on their own to reveal the identity of non-logistic growth processes. They can yield estimates of density compensation, however, which may suggest time lags in density dependence. Analysis of an empirical time series illustrates the issues. PMID- 18395765 TI - Teratogen-induced distortions in the classical NF-kappaB activation pathway: correlation with the ability of embryos to survive teratogenic stress. AB - Studies with diverse teratogens implicated the transcription factor NF-kappaB in mechanisms determining teratological susceptibility of embryos. Here, a teratogen such as cyclophosphamide (CP) was used to test whether teratogenic insult alters the classical NF-kappaB activation pathway, and how these alterations correlate with the ability of mouse embryos to resist the teratogen-induced process of maldevelopment. We observed that embryos tested 24 h after the exposure of females to 40 mg/kg CP exhibited a dramatic decrease in the level of NF-kappaB (p65 subunit)-DNA binding, IkappaB kinase beta (IKKbeta) activity, expression of p65 and IKKbeta proteins, as well as NF-kappaB inhibitory proteins (IkappaBs) such as IkappaBalpha, IkappaBbeta, and IkappaBepsilon, and died within the next 24 h. Embryos of females exposed to 15 mg/kg CP exhibited only a decrease in NF kappaB-DNA binding and IKKbeta activity at 24 h. However, at 48 h, a more prominent decrease in NF-kappaB activity was observed, accompanied by a decreased expression of p65 and IKKbeta proteins. These embryos died within the next 24 h. After treatment with 10 mg/kg CP, embryos survived until the end of the antenatal period of development, demonstrating a transient decrease in NF-kappaB-DNA binding activity and no alterations in NF-kappaB signaling. These results suggest that the classical NF-kappaB activation pathway may be among targets that teratogens engage to initiate abnormal development. Besides, the observation that embryos destined to be dead exhibited a dramatically decreased rate of cell proliferation suggests a pathway, whereby teratogen-induced alterations in NF kappaB signaling may culminate in such a final effect as embryonic death. PMID- 18395766 TI - Protection against heat and staurosporine mediated apoptosis by the HSV-1 US11 protein. AB - US11 protein, one of herpes simplex virus type 1 (HSV-1) true late gene products, plays a role in the virally induced post-transcriptional control of gene expression. In addition, US11 expression also interferes with the cellular response to HSV-1 infection that can lead to apoptosis. We have previously shown that US11 expression enhanced the recovery of cellular protein synthesis and increased cell survival in response to thermal stress. Since heat shock can activate apoptosis, we tested for a possible anti-apoptotic behavior of US11. Here, we show that, in HeLa cells, US11 expression strongly reduced heat induced apoptosis, a phenomenon independent of Hsp expression and characterized by a delayed cytochrome c efflux from mitochondria and reduced caspase 3 activation. Moreover, US11 expression also protected against staurosporine induced apoptosis. Hence, our results favor an anti-apoptotic activity of US11 polypeptide that appears to be located at the level of mitochondria or upstream signaling pathways. PMID- 18395767 TI - Intertrial target-feature changes do not lead to more distraction by singletons: target uncertainty does. AB - The presence of an irrelevant singleton disrupts search for a singleton target substantially more when the target feature varies unpredictably (mixed-singleton search) than when it is known in advance (fixed-singleton search). This finding suggests that advance knowledge of the target feature guides singleton search. Pinto et al. [Pinto, Y., Olivers, C. N. L., & Theeuwes, J. (2005). Target uncertainty does not lead to more distraction by singletons: Intertrial priming does. Perception & Psychophysics, 67, 1354-1361] proposed an alternative account, according to which this difference results from inter-trial priming effects. They based their argument on the finding that distractor interference is reduced when the singleton target feature repeats vs. switches from one trial to the next. However, Lamy et al. [Lamy, D., Carmel, T., Egeth, H., & Leber, A. (2006). Effects of search mode and inter-trial priming on singleton search. Perception & Psychophysics, 68, 919-932] reported no such modulation of distractor interference by target-feature repetition. Here, we show that differences in design (blocking conditions of distractor presence in the former study vs. randomly mixing them in the latter) account for this discrepancy. We conclude that the different task demands induced by the blocked distractor-present and distractor-absent conditions rather than distractor presence per se interact with intertrial priming effects. These findings argue against the claim that singleton search relies exclusively on stimulus-driven factors and suggest that preknowledge of the target feature, when available, can guide attention. In addition, the present results challenge the ambiguity hypothesis of intertrial priming, according to which increased competition for attentional selection boosts inter-trial priming effects. PMID- 18395768 TI - Binocular adaptation to near addition lenses in emmetropic adults. AB - Near addition lenses are prescribed to pre-presbyopic individuals for treatment of binocular motor problems such as convergence excess and to control the progression of myopia. To date, no investigation has looked at the complete sequence of binocular motor responses during a period of near work with +2D lenses. This investigation evaluated changes to accommodation and vergence responses when young adults sustained fixation at 33 cm with +2D addition lenses. In addition, the effect of the accommodative vergence cross-link (AV/A) on the magnitude and the completeness of binocular adaptation to these lenses were evaluated. The results showed that +2D lenses initiate an increase in exophoria and convergence driven accommodation. The degree of the initial induced phoria was dependant upon the magnitude of the AV/A ratio. Vergence adaptation occurred after 3 min of near fixation and reduced the exophoria and convergence driven accommodation. The magnitude of vergence adaptation was dependant upon the size of the induced phoria and hence the AV/A ratio. The completeness of adaptation was seen to vary inversely with induced exophoria and thus the AV/A ratio. PMID- 18395769 TI - Enhanced coagulation of ferric chloride aided by tannic acid for phosphorus removal from wastewater. AB - Phosphorus removal from wastewater is of great importance. In the present study, ferric chloride was selected as the coagulant, and tannic acid (TA), a natural polymer, as the coagulant aid to develop an effective coagulation process with the emphasis of phosphorus recovery from different types of wastewater. The results showed that TA can accelerate the settling speed by forming flocs with large size, reduce the residual Fe(III) to eliminate the yellow color caused by Fe(III), and slightly increase the phosphorus removal efficiency. The precipitate formed by TA-aided coagulation showed the advantage of releasing phosphorus faster than ferric phosphate, indicating the possibility of phosphorus recovery from wastewater as slow release fertilizer. To further understand the structural characteristics of the precipitate, analytical techniques such as Raman spectroscopy, X-ray photoelectron spectroscopy and matrix-assisted laser desorption ionization-time of flight mass spectrometry were employed. The analytical results indicated that TA-Fe-P complex was formed during the coagulation/flocculation processes. Solid phase in the precipitate consisted of TA-Fe-P complex, Fe-TA complex and/or ferric hydroxyphosphate. PMID- 18395770 TI - The primate subarcuate fossa and its relationship to the semicircular canals part II: adult interspecific variation. AB - Studies have reported an empirical link between the size of the semicircular canals and locomotor agility across adult primates. In this paper, we investigate the possibility that this relationship does not follow from the function of the semicircular canals to sense head rotations, but rather reflects spatial constraints imposed by the subarcuate fossa. The latter sits among the three canals and contains the petrosal lobule of the cerebellar paraflocculus, a structure involved in neural processing of locomotion-related eye movements. Hence, it is feasible that agility-related variations of lobule and fossa size affect the arc size of the surrounding semicircular canals. The present study tests such hypothetical correlations by evaluating canal size, fossa size, and agility among extant adult primates. Phylogenetically informed multivariate regression analyses show that, after controlling for body mass, the size of the subarcuate fossa has a significant positive effect on the overall size of the anterior canal and the width of the posterior canal. Multivariate regressions involving the height of the posterior canal and overall size of the lateral canal are not significant. Further bivariate analyses confirm that fossa size is unlikely to play a role in the previously reported link between agility and the size of the posterior and lateral canals. However, fossa size, especially its opening though the arc of the anterior canal, cannot be excluded as a factor that influences the size of the anterior canal more than agility. The findings show that the most reliable functional signals pertaining to locomotion in species that possess a patent subarcuate fossa are likely to come from the lateral canal and are least likely to come from the anterior canal. PMID- 18395771 TI - Lipid profiles in patients with rheumatoid arthritis: mechanisms and the impact of treatment. AB - OBJECTIVE: To describe the impact of rheumatoid arthritis (RA), and its treatment, on lipoprotein levels with potential implications for atherosclerosis. METHODS: A PubMed literature search was undertaken for studies published between 1990 and May 2007, using the search terms "rheumatoid arthritis" AND "lipid" OR "lipoprotein," and including all relevant drug treatment terms for glucocorticoids, disease-modifying antirheumatic drugs, and biologics. RESULTS: Patients with RA face an increased risk of developing premature cardiovascular disease and limited ability to modify risk factors, eg, through exercise. RA is associated with an abnormal lipoprotein pattern, principally low levels of high density lipoprotein (HDL) cholesterol. Most treatments for RA tend to improve the atherogenic index (total/HDL cholesterol ratio), with more evidence for biologics in this regard. The improvement in the lipoprotein profile in RA appears to be associated with suppression of inflammation. CONCLUSIONS: Lipid levels should be monitored and managed in patients with RA to minimize the long-term risk of cardiovascular disease. More research is needed to quantify the relationship between systemic inflammation and lipoprotein levels and to determine the impact of specific lipoprotein particles, eg, small dense low-density lipoprotein and subfractions of HDL on long-term risk. Control of inflammation may have an effect on modifying cardiovascular risk. PMID- 18395772 TI - Endothelial dysfunction, carotid intima-media thickness, and accelerated atherosclerosis in rheumatoid arthritis. PMID- 18395773 TI - Risk factors associated with different stages of atherosclerosis in Colombian patients with rheumatoid arthritis. AB - OBJECTIVES: Rheumatoid arthritis (RA) is associated with an increased prevalence of cardiovascular disease (CVD). Since atherosclerosis development is a gradual process of damage inside the artery wall, and the phenotype-genotype correlation of complex diseases may vary depending on ethnicity, we sought to investigate the influence of clinical features, routine inflammatory markers, and the genetic component of RA on different stages of atherosclerosis in northwestern Colombian patients with RA. METHODS: A group of 140 patients with RA were enrolled in this study. All patients underwent a noninvasive evaluation of endothelial function by flow-mediated vasodilation (FMV) and an assessment of carotid intima-media thickness (IMT) by high-resolution B-mode ultrasonography. The patients were classified into 3 categories: endothelial dysfunction (FMV <5%), increased IMT (0.91-1.29 mm), and plaque (IMT >1.30 mm). The risk of being in each category was assessed by investigating traditional and nontraditional cardiovascular risk factors. For each stage of atherosclerosis development, we searched for nontraditional risk factors that were significantly associated with the stage after adjusting for traditional risk factors and current age. RESULTS: Rheumatoid factor seropositivity was significantly associated with endothelial dysfunction (adjusted odds ratio, AOR = 3.0). A duration of RA >10 years (AOR = 29.0) and being a carrier of an HLA-DRB1 shared epitope allele (AOR = 4.8) were associated with atherosclerotic plaque. No association of extra-articular manifestations, anticyclic citrullinated peptide (anti-CCP3) antibodies, and tumor necrosis factor -308 polymorphism with CVD was found. CONCLUSIONS: Our results reveal the presence of RA-related risk factors for CVD which act independently of traditional risk factors. These factors can be used by clinicians to predict CVD in RA patients, and this data should assist in the development of public health policies in our population for the improvement of patient outcomes. PMID- 18395774 TI - Insertion of inferior vena cava filters in patients with the antiphospholipid syndrome. AB - BACKGROUND: The antiphospholipid syndrome (APS) is a disease with a high prevalence of thromboembolic events, especially pulmonary emboli (PE). These events may recur despite anticoagulation therapy. In such cases, placement of an inferior vena cava (IVC) filter may be considered to prevent propagation of a distal thrombus toward the pulmonary vessels. It is unclear whether the placement of such a filter is beneficial in patients with APS. OBJECTIVE: Retrospective evaluation of the value of IVC filter placement as prophylaxis against recurrent pulmonary emboli in patients with medically treated APS. METHODS: We identified 10 patients suffering from APS who, despite anticoagulation treatment, had recurrent thromboembolic events. All of them underwent placement of an IVC filter. We examined their medical files for further recurrences. RESULTS: Of the 10 patients in our study, only 1 had a documented PE following the intervention. The remaining patients had no evidence of PE after the filter insertion. Five of the 10 patients died, 2 of them suddenly. In those 2 patients, the cause of death is unknown, but PE cannot be excluded. CONCLUSION: IVC filters seem to be protective against recurrent PE in APS patients but the true extent of their efficacy requires further study. PMID- 18395775 TI - High adiponectin and adiponectin receptor 1 expression in synovial fluids and synovial tissues of patients with rheumatoid arthritis. AB - OBJECTIVES: The major objectives of this article are first to measure the levels of expression of adiponectin and its 2 receptors (adipoR1 and adipoR2) in the peripheral blood mononuclear cells and in the synovial compartment of rheumatoid arthritis (RA) patients, and second, to assess their pro-inflammatory potential. Osteoarthritis patients and healthy subjects served as controls. METHODS: Expression of adiponectin, adipoR1, and adipoR2 were assayed by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistology. The potential pro-inflammatory activity of adiponectin was studied by adding recombinant adiponectin to cultures of synovial fibroblasts. RESULTS: Immunohistology showed that numerous cells in the synovial biopsies of RA expressed adiponectin, adipoR1, and adipoR2. The synovial fibroblasts were distinctly rich in adiponectin. As expected, high adiponectin levels were present in the synovial fluids. In contrast to the synovial compartment, in peripheral blood mononuclear cells, only adipoR1 exceeded those of osteoarthritis and healthy subjects. When recombinant adiponectin was added to cultures of synovial fibroblasts, it induced up to 8.1- and 11.4-fold increase in the release of monocyte chemoattractant protein-1 and interleukin-6. CONCLUSIONS: The adiponectin adipokine axis might play a role in RA. PMID- 18395777 TI - Do clear cell ovarian carcinomas have poorer prognosis compared to other epithelial cell types? A study of 1411 clear cell ovarian cancers. AB - OBJECTIVE: To compare the clinico-pathologic characteristics and survival of women with clear cell versus other epithelial ovarian cancers. METHODS: Data were obtained from the Surveillance, Epidemiology and End Results Program between 1988 and 2001 and analyzed using Kaplan-Meier and Cox proportional hazards models. RESULTS: Of 28,082 women with epithelial ovarian cancer, 1411 (5%) had clear cell, 13,835 (49.3%) papillary serous, 3655 (13%) endometrioid, 2711 (9.7%) mucinous, and 6470 (23%) had unspecified histologies. The median age of overall patients was 64 years; with clear cell patients presenting at younger age (55 years). The proportion of clear cell histology was significantly higher in Asians versus Whites, Blacks, and others (11.1% versus 4.8%, 3.1%, and 5.5%; p<0.001). Clear cell carcinoma is more likely to be diagnosed at early-stage (67.3%) compared to 19.2% in serous, 61.6% endometrioid, and 61.3% in mucinous carcinomas (p<0.005). Retroperitoneal lymph node metastases were found in 13.6% of serous carcinomas, 7.9% clear cell, 7.3% endometrioid, and 3.8% of mucinous (p<0.001). Adjusted for stage, the 5-year disease-specific survival of patients with clear cell carcinoma is worse compared to serous: 85.3% vs. 86.4% for stage I, 60.3% vs. 66.4% stage II, 31.5% vs. 35.0% stage III, and 17.5% vs. 22.2% for stage IV, respectively (p<0.001). On multivariate analysis, age, stage, grade, histology, and surgical treatment were independent predictors of disease-specific survival. CONCLUSIONS: Our data suggest that women with clear cell ovarian cancer present at a younger age, are more likely to be Asian, and have a poorer prognosis compared to serous cancers. PMID- 18395778 TI - Stage IV endometrial cancer after failed conservative management: a case report. AB - BACKGROUND: With younger women becoming increasingly overweight, endometrial cancer is becoming a concern in this group. These women are likely to desire fertility preservation. There are no randomized controlled data to guide conservative therapy. CASE: A 24-year-old female with endometrial adenocarcinoma diagnosed in December 2003 was treated conservatively with Megace and levonorgestrel intrauterine device. She was lost to follow-up and represented in March 2006. Endometrial biopsy revealed grade 1 endometrial adenocarcinoma. Definitive surgical therapy was recommended, however, the patient declined surgery in the interest of preserving fertility until November 2006. Final pathology revealed a stage IV, grade 1 endometrioid endometrial cancer. CONCLUSION: Women with endometrial cancer who desire fertility preservation should be counseled regarding the possible risk of advanced disease if surgical therapy is delayed. PMID- 18395779 TI - Potential role of Renin-Angiotensin-system for tumor angiogenesis in receptor negative breast cancer. AB - OBJECTIVE: This study examined the potential role of Angiotensin II for the regulation of angiogenesis associated genes in receptor positive and negative human breast cancer. METHODS: Expression of different Renin-Angiotensin system (RAS) components in human breast cancer tissue was investigated using immunofluorescence, and in a receptor positive (MCF-7) and receptor negative (MDA MB 468) breast cancer cell line by performing immunocytochemistry and RT-PCR. Both cell lines were stimulated with Angiotensin II and Angiotensin II receptor type 1 (At(1)R) blocker Candesartan, and gene expression of vascular endothelial growth factor (VEGF), Angiopoietin 1 and 2 (Ang-1 and Ang-2), tissue inhibitor of matrix metalloproteinases 1 (TIMP-1), and hypoxia inducible transcription factor 2alpha (HIF-2alpha) were quantified by TaqMan-Real-Time PCR analysis. RESULTS: RAS components, Angiotensinogen, Renin, Angiotensin I-converting enzyme (ACE), and At(1)R and At(2)R were expressed in hormone-receptor negative and positive human breast cancer tissue as well as in MDA-MB 468 and in MCF-7 human breast cancer cells. In addition, we found expression of VEGF, Ang-1, TIMP-1, and HIF 2alpha in both cell lines. However, only in receptor negative MDA-MB 468 cells, did Angiotensin II significantly increase gene expression of VEGF, HIF-2alpha, and TIMP-1. This effect was completely inhibited by Candesartan. CONCLUSION: In conclusion, it is hypothesized that Angiotensin II may be involved in regulation of tumor angiogenesis especially in receptor negative breast cancer by regulation of angiogenesis associated genes via At(1)R. These findings are the first evidence for targeting tumor angiogenesis by inhibition of At(1)R in receptor negative human breast cancer cells and may lead to new therapeutical anticancer strategies based upon inhibition of At(1)R. PMID- 18395780 TI - Novel discriminative stimulus effects of TPA023B, subtype-selective gamma aminobutyric-acid(A)/benzodiazepine modulator: comparisons with zolpidem, lorazepam, and TPA023. AB - Anxiolytics with fewer unwanted effects may be created by varying GABAergic efficacy at the BZ binding site across GABA(A) receptor subtypes. TPA023 and TPA023B have in vitro antagonist efficacy at alpha(1) subtypes and partial agonist efficacy at alpha(2/3) subtypes. TPA023B has partial-agonist efficacy at alpha(5); TPA023 has none. Drug discrimination procedures were used to determine whether the novel GABA(A) receptor efficacy profiles would be reflected in a model of subjective effects of BZ-site ligands. Rats were trained to discriminate TPA023, TPA023B, the nonselective BZ anxiolytic lorazepam, or the alpha(1) selective hypnotic zolpidem. The lorazepam, zolpidem, and TPA023 discriminations were learned in < 50 sessions. The TPA023B training group showed no evidence of acquiring the TPA023B discrimination after 160 sessions despite various procedural manipulations. Neither zolpidem- nor lorazepam-trained rats generalized to TPA023B. Within the same dose range, however, TPA023-trained rats generalized fully and dose-dependently to TPA023B. Number of training sessions to regain criterion discrimination performance following TPA023B tests in the lorazepam, zolpidem, and TPA023 groups increased as a function of dose, likely due to effects of residual TPA023B. Together with previous data, the present results suggest that elimination of alpha(1) efficacy plus reductions in alpha(2/3) efficacy permits anxiolysis but decreases BZ-like interoceptive stimulus effects. PMID- 18395781 TI - Questioning the economic evaluation of omalizumab. PMID- 18395782 TI - Airborne allergy to wheat lipid transfer protein without food allergy? PMID- 18395783 TI - Filaggrin variants confer susceptibility to asthma. PMID- 18395784 TI - Gender and psychotropic medication use: the role of intimate partner violence. AB - OBJECTIVE: Women are known to use more psychotropic medications than men which may be linked to women's greater exposure to intimate partner violence (IPV). METHOD: The use of medications for sleep, depression and anxiety in adults in the 1999 Canadian General Social Survey was assessed. Rates of medication use by adults exposed to IPV (physical, sexual, emotional and financial) were compared to rates of those reporting no IPV. RESULTS: More women (14.9%) than men (9.6%) reported use of psychotropic medications in the preceding month. Rates were significantly higher in both women and men who reported IPV. This link was still present after key sociodemographic and health predictors of medication use were held constant. CONCLUSION: This random population based study provides the first data to support the idea that IPV may explain at least some of the increased psychotropic medication use by women. IPV should be included as a predictor variable in future studies investigating psychotropic medication use which itself can be added to the list of adverse health risks of IPV. PMID- 18395785 TI - Effect of mouth-motion fatigue and thermal cycling on the marginal accuracy of partial coverage restorations made of various dental materials. AB - OBJECTIVES: To investigate the influence of mouth-motion fatigue on marginal accuracy of partial-coverage-restorations-(PCRs) of various dental materials. METHODS: Eighty molars were prepared equally and divided into five groups (n=16). PCRs were fabricated of following dental materials: Group-GO=Gold-Pontor MPF(double dagger), Group-TA=Targis*, Group-EX=IPS-e.max-Press*, Group-EM=IPS Empress*, Group-PC=ProCAD*/Cerec 3(dagger) ((double dagger)Metalor/*Ivoclar Vivadent/(dagger)Sirona-Dental-System). Gold-PCRs were cemented conventionally. Residual 64 PCRs were adhesively luted and subjected to masticatory loading (1.2million-cycles, 1.6Hz, 49N) and thermal cycling (5 degrees C/55 degrees C, 60s, dwell-time, 5500cycles). Discrepancies in marginal-accuracy were examined on epoxy replicas (200 x magnification). Statistical analysis was performed by unpaired and paired t-tests (alpha=0.05). RESULTS: After cementing, marginal accuracy (geometrical mean)[95% confidence limits] was recorded: GO-47[43 51]microm, TA-42[38-45]microm, EX-60[52-67]microm, EM-52[45-60]microm and PC 75[59-94]microm. No significant differences were found between groups GO, TA and EM. Values of Group-EX were significantly higher compared to Group-TA (p=0.04). Group-PC demonstrated significantly decreased marginal-accuracy towards groups GO (p=0.03) and TA (p=0.02). Except for Group-GO (p=0.01), no significant changes in marginal-accuracy were observed after mouth-motion fatigue and thermal cycling (GO-42[38-45]microm, TA-42[38-47]microm, EX-56[49-65]microm, EM-54[46-64]microm and PC-71[59-84]microm). However, Group-GO and Group-EM showed significant deviations in marginal-accuracy after aging (p=0.04). Marginal discrepancies of groups EX and EM were similar (p=1.0). Values of Group-PC were significantly higher when compared to groups GO (p=0.01) and TA (p=0.02). Buccal-lingual marginal discrepancies were significantly higher than mesial-distal in all groups and stages. SIGNIFICANCE: Cast-gold-PCRs demonstrated superior marginal-accuracy, followed by indirect ceromer PCRs. All-ceramic PCRs showed in vitro clinical acceptable marginal widths, even though CAD/CAM fabrication slightly compromised marginal-accuracy. Since preparation margins were not exposed to occlusal loading directly, increased marginal discrepancies due to marginal chipping during mouth motion fatigue were not experienced. PMID- 18395786 TI - Cloning a new cytochrome P450 isoform (CYP356A1) from oyster Crassostrea gigas. AB - We have cloned the full-length cDNA of the first member of a new cytochrome P450 (CYP) family from the Pacific oyster Crassostrea gigas. This new CYP gene was obtained based on an initial 331bp fragment previously identified among the list of the differentially expressed genes in oysters exposed to untreated domestic sewage. The full-length CYP has an open reading frame of 1500bp and based on its deduced amino acid sequence was classified as a member of a new subfamily, CYP356A1. A phylogenetic analysis showed that CYP356A1 is closely related to members of the CYP17 and CYP1 subfamilies. Semi-quantitative RT-PCR was performed to analyze the CYP356A1 expression in different tissues of the oyster (digestive gland, gill, mantle and adductor muscle). Results showed slightly higher CYP356A1 expression in digestive gland and mantle, than the other tissues, indicating a possible role of the CYP356A1 in xenobiotic biotransformation and/or steroid metabolism. PMID- 18395787 TI - Biochemical alterations in juvenile carp (Cyprinus carpio) exposed to zinc: glutathione reductase as a target. AB - The aim of this study was to investigate biochemical changes in juvenile carp (Cyprinus carpio) exposed to zinc chloride (10, 30 and 100 microM) for a period of 48 h. Zinc exposure caused a concentration-dependent reduction in glutathione reductase (GR) activity in gills, liver and brain. Gill glutathione S-transferase (GST) was reduced when animals were exposed to the highest concentration of 100 microM zinc. The phosphorylation of p38(MAPK) increased in the brain of fish exposed to zinc 100 microM, while phosphorylation of the extracellular signal regulated protein kinase 1/2 (ERK1/2) and c-Jun N-terminal protein kinase 1/2 (JNK1/2) remained unchanged. Expression of proteins HSP60 and HSP70 were not affected by zinc exposure. Considering the significant concentration-dependent inhibition of GR in all tissues analyzed, this enzyme could be a potential biomarker of exposure to zinc, which has to be confirmed. PMID- 18395788 TI - Differential gene expression in oyster exposed to sewage. AB - In order to investigate the influence of domestic sewage on the gene expression of Pacific oysters Crassostrea gigas, suppression subtractive hybridization (SSH) method was employed. Oysters were sampled at a farming area and, after 10 days of acclimation in the laboratory, were exposed to untreated domestic sewage diluted 33% for 48 h. Gills of male oysters were excised for total RNA extraction. mRNA was purified and the differential gene expression was analyzed by SSH. We obtained 61 cDNA sequences but only 15 were identified, which includes fatty acid binding protein, multidrug resistance protein, omega glutathione S-transferase, cytochrome P450 isoform CYP356A1, among others. The identified genes are associated with different metabolic functions like biotransformation, membrane transport, aerobic metabolism and translational machinery, evidencing the potential toxic effects elicited by these effluents. PMID- 18395789 TI - In vitro study of the effects of cadmium on the activation of the estrogen response element using the YES screen. AB - Estrogenic potential of environmental samples is frequently assessed using receptor-based functional assays. Using the yeast estrogen screen (YES) developed by Routledge and Sumpter, we assessed the ability of cadmium to activate the estrogen receptor-mediated response. No induced transcriptional activity was observed with a range of CdCl2 concentrations (1 nm-1mM). But, when combining cadmium with the model compound 17beta-estradiol, cadmium was able to significantly potentiate the induced estrogenic response for concentrations ranging from 15 nM to 1 microM. A maximal effect was observed at 0.5 microM with a ten fold reduction of the 17beta-estradiol EC50. PMID- 18395790 TI - Comparison of the antioxidant defense system in Crassostrea rhizophorae and Crassostrea gigas exposed to domestic sewage discharges. AB - Oysters Crassostrea rhizophorae and Crassostrea gigas were kept for fourteen days at four sites in Sao Jose, SC, Brazil, chosen according to a sewage discharge contamination gradient. Enzymatic activities of CAT, GST, G6PDH and GR were evaluated in gills and digestive glands. Higher CAT activity was observed in tissues of C. rhizophorae in response to sewage contamination possibly indicating peroxisome proliferation induction. C. gigas showed elevated G6PDH activity in digestive gland, and GR in gills, after domestic sewage exposure. In conclusion, C. rhizophorae and C. gigas showed different biochemical responses after in situ exposure to domestic sewage. However, C. rhizophorae showed more significant changes in CAT suggesting that this organism could be a better monitor to this kind of effluent. PMID- 18395791 TI - Keratin transamidation. AB - Low molecular weight keratin was self-crosslinked by microbial transglutaminase (mTGase) for application to wool fabric. From amino acid determination, keratin produced by the alkaline hydrolysis of wool showed requisite glutamine and lysine required for mTGase-mediated transamidation. Keratin showed less lysyl amine content after combination with mTGase as proof of self-crosslinking. Gel electrophoretic patterns provided evidence of self-crosslinking with the development of relatively higher molecular weight protein bands within 30 min after mTGase was combined with keratin at 30 degrees C. Increase in the deconvoluted amide II band from IR spectra of keratin after combination with mTGase provided further evidence of transamidation. By examining the ability of keratin to self-crosslink, past findings were elucidated whereby mTGase-mediated crosslinking imparted strength to wool and keratin controlled its dimensional stability. mTGase-catalyzed isopeptide bond formation of keratin to form monosubstituted gamma-amides of peptide-bound glutamine in epsilon-amino-(gamma glutamyl)lysine crosslinks. This system was effective for binding wool to wool, keratin to wool, and keratin to keratin in self-crosslinking. PMID- 18395792 TI - A synthetic amyloid lawn system for high-throughput analysis of amyloid toxicity and drug screening. AB - Amyloid-beta (Abeta) is the major constituent of senile plaques in the brains of Alzheimer's disease patients. In order to develop an efficient in vitro system for studying the interaction of cells with Abeta aggregates, we have prepared a synthetic amyloid lawn by immobilizing Abeta peptides over a functionalized glass surface and subsequently incubating the template in a fresh Abeta solution. On the top of different types of amyloid lawns (e.g. monomeric, oligomeric, and fibrillar), we cultivated PC12 cells, creating physical contacts between the cells and the lawns. Results indicated that cell viability was differentially affected when grown atop different Abeta lawns while cells were well adhered onto the surface of these Abeta lawns. The mode of cell death by Abeta lawn was confirmed to be apoptotic rather than necrotic, showing that cells undergo suicide by just contact with Abeta lawn. While conventional 'solution-based' methods for testing amyloid toxicity suffer from problems such as lot-to-lot variations, continued fibrillation, and heterogeneous population of aggregates, our 'surface-based' lawn system is suitable for high-throughput analysis of amyloid toxicity, which may enable high-throughput screening of potential drug candidates for treating amyloid diseases with the goal of reducing the cell death on the lawn. PMID- 18395793 TI - Immobilization of urokinase on the islet surface by amphiphilic poly(vinyl alcohol) that carries alkyl side chains. AB - Transplantation of islets of Langerhans (islets) is a promising method to treat insulin-dependent diabetes mellitus (type I diabetes). However, insulin independence is typically realized for only approximately 30% of transplant recipients, even with sufficient numbers of islets from multiple donors. Innate immunological reactions triggered by blood coagulation play a key role in the loss of islets at the early stage. Here we propose a method to inhibit blood coagulation on the islet surface. A plasminogen activator, urokinase, was immobilized on the islet surface via a poly(vinyl alcohol) (PVA) derivative that carries alkyl chains and thiol groups. When the PVA derivative was added to an islet suspension, the alkyl side chains spontaneously anchored into the lipid bilayer membranes of islet cells. The surfaces of islets were covered with the PVA derivative. Urokinase modified with maleimide groups could be immobilized onto the islet surface by thiol/maleimide bonding with the layer of PVA derivatives. Urokinase-immobilized islets exhibited fibrinolytic properties, indicating that blood coagulation can be controlled on the islet surface. Urokinase immobilization on islets, which does not impair insulin release, represents a promising method to reduce early graft loss after intraportal islet transplantation. PMID- 18395794 TI - Antioxidant gene expression in preeclamptic placentae: a preliminary investigation. AB - Oxidative stress has been implicated in the pathogenesis of preeclampsia. This study measured the relative mRNA expression of antioxidant proteins glutathione peroxidase 1 and 4, glutathione reductase, thioredoxin 1 and 2, thioredoxin reductase 1, thioredoxin peroxidase 3 and superoxide dismutase 1 and 2 in preeclamptic and non-preeclamptic placentae. Quantitative real-time PCR was conducted on placental mRNA isolated from preeclamptic and control patients. Cycle threshold numbers and fold differences were calculated as a measure of linear product amplification and used for comparison. The mRNA expression of glutathione reductase was significantly reduced (fold difference 0.41, p<0.05) in preeclamptic placenta when compared to controls while the expression of thioredoxin peroxidase 3 was significantly increased (fold difference 3.25, p<0.001) in the preeclamptic placentae. No significant difference in expression was observed for glutathione peroxidase 1 and 4, thioredoxin 1 and 2, thioredoxin reductase 1 and superoxide dismutase 1 and 2. These results suggest that it is the abnormal oxidative insult associated with preeclampsia not mRNA expression of antioxidant proteins that may be responsible for reduced antioxidant enzyme activity in preeclamptic placentae. PMID- 18395795 TI - Detection of elastin derived peptides in cerebrospinal fluid of patients with first ever ischaemic stroke. AB - OBJECTIVE: We have previously reported the optimized methods for the detection of elastin derived peptides (EDP) in the serum, synovial fluid, and bronchoalveolar lavage. The aim of the present study was to investigate whether EDP are detectable in cerebrospinal fluid (CSF) of patients with acute brain ischaemia. PATIENTS AND METHODS: Twenty-seven first ever ischaemic stroke patients (mean age 61.5+/-10.8 years; age range 47-70 years; 12 women) were studied in acute phase (1-15 days after the onset) with clinical evaluations, radiological assessments, and the analysis of serum and CSF based on Western blot and ELISA for the detection and quantification of EDP. RESULTS: None of the serum EDP concentrations are significantly higher in stroke patients compared with 25 healthy control individuals. However, EDP levels in CSF are strongly (p<0.0001) elevated compared with healthy subjects. They correlated with total cholesterol (r=0.53; p=0.02), triglycerides (r=0.67; p=0.004) and retinopathy (r=0.24; p=0.03), and with the interval between the stroke onset and the time of lumbar puncture (r=0.35; p=0.02). CONCLUSION: EDPs are detectable in CSF of healthy subjects and patients with ischaemic stroke. Acute brain infarction is followed by increased levels of EDP in CSF. PMID- 18395796 TI - Rapid accumulation of an interleukin 17 homolog transcript in Crassostrea gigas hemocytes following bacterial exposure. AB - Interleukin-17 (IL-17) gene models have been found in the sequenced genomes of Strongylocentrotus purpuratus and Caenorhabditis elegans. However, there have been no published reports on the empirical cloning and characterization of any interleukin cDNAs in invertebrates. From a Pacific oyster (Crassostrea gigas) hemocyte cDNA library, two clones were obtained that encoded a protein similar to vertebrate IL-17s. The putative oyster IL-17 homolog (CgIL-17) was 27% identical to rainbow trout IL-17D, 21% to human IL-17D and 24% to an IL-17D-like gene model obtained from the annotation of the sea urchin genome. IL-17s from the oyster, sea urchin, trout and human, contained conserved cysteine residues found in all forms of IL-17 in mammals. Injection of bacteria into C. gigas oysters produced a large and rapid elevation in CgIL-17 transcript abundance in hemocytes, suggesting that this is a very early response gene to pathogens that may be responsible for the stimulation of other immune genes in the oyster. PMID- 18395797 TI - Expression of three serine protease genes from the South East Asian malaria vector, Anopheles dirus, in relation to blood feeding and parasite infection. AB - Three serine protease cDNA clones were isolated from Anopheles dirus, a major vector of malaria in Southeast Asia. Transcript abundance was examined following infection by Plasmodium falciparum by RT-PCR analysis. SerF3 exhibited increased transcript abundance in the whole body at 10 days post-infection with P. falciparum. All three genes are candidates for further investigation to determine their roles in mosquito immune responses. PMID- 18395798 TI - A method for generating sticky-end PCR products which facilitates unidirectional cloning and the one-step assembly of complex DNA constructs. AB - We have developed and tested a method for the restriction enzyme-independent generation of sticky-end PCR products. The method is suitable for use with a proof-reading polymerase such as pfu, or any other heat-stable polymerase which produces a blunt-end product. The technique can be used to achieve unidirectional cloning of PCR products with an efficiency greater than 90%. Because the sequences of the sticky ends are defined by the user and potentially can be of any length, the method can also be exploited for the one-step construction of recombinant plasmids from multiple functional cassettes, without the use of restriction enzymes. PMID- 18395799 TI - Foxp3 induces IL-4 gene silencing by affecting nuclear translocation of NFkappaB and chromatin structure. AB - The forkhead family protein Foxp3 is a unique marker of regulatory T cells and plays a crucial role in the development and function of those cells. Ectopic expression of Foxp3 abolishes the expression of many cytokines in uncommitted cells but there is little information about whether it causes gene silencing in differentiated cells. In this study, we showed that ectopic expression of Foxp3 in primary T helper 2 cells abolished IL-4 gene expression. Foxp3 inhibited nuclear translocation of NFkappaB by increasing the stability of the NFkappaB inhibitor IkappaBalpha, which in turn reduced in vivo binding of NFkappaB to the IL-4 promoter region. Moreover, Foxp3 over-expression induced inactive chromatin structure by decreasing in vivo binding levels of acetylated histone 3 while increasing methylated histone 3 at lysine 9 in the IL-4 genomic locus. Our results suggest that Foxp3 could induce gene silencing by inhibiting NFkappaB activity and by causing its target loci to adopt an inactive chromatin configuration. PMID- 18395800 TI - Clinical course of retrobulbar hemangioblastomas in von Hippel-Lindau disease. AB - OBJECTIVE: To report clinical findings of rare retrobulbar optic nerve hemangioblastomas associated with von Hippel-Lindau disease (VHL). DESIGN: Retrospective observational case series. PARTICIPANTS: Nine patients with VHL. METHODS: The clinical course and magnetic resonance imaging findings of patients with VHL and hemangioblastomas affecting the anterior visual pathway from the intraorbital optic nerve to the optic chiasm are reviewed. MAIN OUTCOME MEASURE: Clinical course of retrobulbar optic nerve hemangioblastomas. RESULTS: The mean age of VHL diagnosis was 24+/-14 years, and mean follow-up was 5+/-4 years. All had other CNS lesions and retinal hemangioblastomas. Approximately 50% (5/9) had a previous enucleation or had visual acuity loss (4/9), some due to other VHL ocular complications. Four patients underwent surgical resection of an intracranial hemangioblastoma. Growth patterns and pathology are similar to those of other hemangioblastomas in the CNS. CONCLUSIONS: Although these lesions are rare, patients with VHL who present with signs of optic neuropathy should be evaluated for anterior visual pathway hemangioblastomas impinging on the optic nerve from the orbit to the chiasm. On neuroimaging, the hemangioblastomas may demonstrate chiasmal or optic tract edema, associated cysts, and T(2) flow voids. Lesions may remain radiologically and clinically stable, evolve radiographically with no visual or neurological progression, or progress clinically and radiographically. Patients at risk for visual loss should be considered for surgical resection. Close coordination among neuroradiology, neurosurgery, and ophthalmology patient care teams is advised for optimal management of these patients. PMID- 18395801 TI - Treatment of cats with feline infectious peritonitis. AB - Feline infectious peritonitis (FIP) infection resulting in clinical signs is invariably fatal despite clinical intervention. As FIP is an immune-mediated disease, treatment is mainly aimed at controlling the immune response triggered by the infection with the feline coronavirus (FCoV). Immune suppressive drugs such as prednisone or cyclophosphamide may slow disease progression but do not produce a cure. In nearly every published case report of attempted therapy for clinical FIP, glucocorticoids have been used; there are, however, no controlled studies that evaluate the effect of glucocorticoids as a therapy for FIP. Some veterinarians prescribe immune modulators to treat cats with FIP with no documented controlled evidence of efficacy. It has been suggested that these agents may benefit infected animals by restoring compromised immune function, thereby allowing the patient to control viral burden and recover from clinical signs. However, a non-specific stimulation of the immune system may be contraindicated as clinical signs develop and progress as a result of an immune mediated response to the mutated FCoV. PMID- 18395802 TI - Cochlear implantation in individuals with Usher type 1 syndrome. AB - OBJECTIVE: To analyze the occurrence of the Usher type 1 (USH1) gene mutations in cochlear implant recipients with deaf-blind Usher syndrome, and to assess the potential effect of these genes and other factors on the therapeutic outcome. STUDY DESIGN: Case series study of nine patients with the phenotypic diagnosis of USH1. METHODS AND SUBJECTS: Mutation analysis of four USH1 genes (MYO7A, USH1C, CDH23, and PCDH15) by single strand conformational polymorphism (SSCP) and direct sequencing methods. Pre- and post-implantation audiologic tests including pure tone audiometry, speech perception measures, and qualitative assessment of auditory performance. Nine USH1 patients who received their cochlear implants at the University of Miami Ear Institute, Miami, FL, USA, and at the Department of Cochlear Implants, Great Ormond Street Hospital for Children, London, UK. RESULTS: DNA samples from five of the nine patients were available for mutation analysis. Three of the five patients were found to carry USH1 mutations including two with a truncated mutation in CDH23 and one being a digenic inheritance with mutations in CDH23 and PCDH15. We may have failed to detect mutations in the amplicons analyzed, as neither SSCP nor direct sequencing, even combined, detects all mutations present. Our failure to detect mutations in all five patients may also confirm the genetic heterogeneity of USH1 and additional USH1 loci remain to be mapped. Pre-implantation assessment indicated that all of the subjects were pre-linguistically profoundly deaf, had no consistent response to sound, had varying degrees of auditory-oral habilitation. Age at implantation ranged from 2 to 11 years. There was post-implantation improvement in sound detection and speech recognition measures in closed-set format in all patients. Children implanted at an age of 3 years or less showed good open-set speech perception with lip-reading. All patients are implant users. Those patients who do not show open-set perception still use the cochlear implant as an adjunct of lip-reading or total communication. CONCLUSION: Testing for mutations in the USH1 genes allows early identification and intervention of children with USH1; timely intervention is important to maximize the development of useful auditory-oral communication skills prior to the onset of the visual impairment. PMID- 18395803 TI - The effect of task type on fundamental frequency in children. AB - OBJECTIVE: The purpose of this study was to examine the influence of task type on the fundamental frequency (F(0)) produced by young children. Fundamental frequency is a parameter which describes the rate of vocal fold vibration. The influence of task type on F(0) values is important for health professionals designing and implementing assessment protocols for children with voice disorders. METHODS: Forty-eight healthy children between the ages of 5.0 and 7.11 years were evaluated in this study. Each child completed four tasks used to elicit a voice sample for subsequent analysis of F(0). The tasks included: (a) sustaining the vowel/a/(Vowel), (b) sustaining the vowel embedded in a word at the end of a phrase (Phrase), (c) repeating a sentence (Sentence), and (d) counting from 1 to 10 (Counting). Each child was evaluated by one of two clinicians who had an equal number of years of training and were the same age and gender. RESULTS: A repeated measures analysis of variance (ANOVA) was used to examine the influence of task type on F(0) values. The results revealed a significant difference in F(0) between the four elicitation tasks (p=.002). Pair wise comparisons revealed that Counting elicited higher F(0) values compared to Phrase (p=.018) and Sentence tasks (p=.001). There were no significant interaction effects for task by age, gender, or clinician who evaluated the child (p>.05). CONCLUSIONS: The results of this study revealed that task type does significantly influence F(0) values in young children. This finding is clinically important as it indicates that the same task should be used to monitor changes in the voice over time in relationship to surgical or behavioral interventions. PMID- 18395804 TI - The use of Armored RNA as a multi-purpose internal control for RT-PCR. AB - Real time reverse transcriptase-PCR (RT-PCR) is now used commonly for the detection of viral pathogens in respiratory samples. However, due to potential inhibition of the RT-PCR or inefficient extraction, this sample type can present significant challenges to accurate patient testing. The goal of this study was to create an internal control to be multiplexed in a real time RT-PCR assay for detecting a viral target in respiratory samples. This report describes an Armored RNA (aRNA) internal control developed originally to be multiplexed in a real time RT-PCR assay for detecting SARS-associated Coronavirus, but can be incorporated into any RT-PCR assay. The internal control primers and probe target a region in the coat protein gene of the E. coli F-specific bacteriophage ms2, which is contained within the aRNA. PMID- 18395806 TI - RNA editing, DNA recoding and the evolution of human cognition. AB - RNA editing appears to be the major mechanism by which environmental signals overwrite encoded genetic information to modify gene function and regulation, particularly in the brain. We suggest that the predominance of Alu elements in the human genome is the result of their evolutionary co-adaptation as a modular substrate for RNA editing, driven by selection for higher-order cognitive function. We show that RNA editing alters transcripts from loci encoding proteins involved in neural cell identity, maturation and function, as well as in DNA repair, implying a role for RNA editing not only in neural transmission and network plasticity but also in brain development, and suggesting that communication of productive changes back to the genome might constitute the molecular basis of long-term memory and higher-order cognition. PMID- 18395805 TI - Circuit-based framework for understanding neurotransmitter and risk gene interactions in schizophrenia. AB - Many risk genes interact synergistically to produce schizophrenia and many neurotransmitter interactions have been implicated. We have developed a circuit based framework for understanding gene and neurotransmitter interactions. NMDAR hypofunction has been implicated in schizophrenia because NMDAR antagonists reproduce symptoms of the disease. One action of antagonists is to reduce the excitation of fast-spiking interneurons, resulting in disinhibition of pyramidal cells. Overactive pyramidal cells, notably those in the hippocampus, can drive a hyperdopaminergic state that produces psychosis. Additional aspects of interneuron function can be understood in this framework, as follows. (i) In animal models, NMDAR antagonists reduce parvalbumin and GAD67, as found in schizophrenia. These changes produce further disinhibition and can be viewed as the aberrant response of a homeostatic system having a faulty activity sensor (the NMDAR). (ii) Disinhibition decreases the power of gamma oscillation and might thereby produce negative and cognitive symptoms. (iii) Nicotine enhances the output of interneurons, and might thereby contribute to its therapeutic effect in schizophrenia. PMID- 18395808 TI - Glial alterations in the hippocampus of rats submitted to ibotenic-induced lesion of the nucleus basalis magnocellularis. AB - Lesion of the nucleus basalis magnocellularis (nbm) is a suitable approach to study cognitive deficit and behavior alterations involving cholinergic dysfunction, which is associated with the major types of dementia. Cortical astrogliosis also has been described in this model, but it is not clear whether hippocampal astrocytes are activated. In this study, we investigated possible specific astrocyte alterations in the hippocampi of Wistar rats submitted to nbm damage with ibotenic acid, investigating the content and immunohistochemistry of glial fibrillary acidic protein (GFAP), as well as S100B protein content, glutamate uptake and glutamine synthetase activity on the 7th and 28th post lesion days. Cognitive deficit was confirmed by the step-down inhibitory avoidance task. Interestingly, we found a decrease in GFAP content, S100B content and glutamate uptake activity in the hippocampus on the 28th day after nbm lesion. No alterations were observed in glutamine synthetase activity or in the cerebrospinal fluid S100B content. Although our data suggest caution in the use of nbm lesion with ibotenic acid as a dementia model, it is possible that these alterations could contribute to the cognitive deficit observed in these rats. PMID- 18395807 TI - Axonal growth therapeutics: regeneration or sprouting or plasticity? AB - Loss of function after neurological injury frequently occurs through the interruption of axonal connectivity, rather than through cell loss. Functional deficits persist because a multitude of inhibitory factors in degenerating myelin and astroglial scar prevent axonal growth in the adult brain and spinal cord. Given the high clinical significance of achieving functional recovery through axonal growth, substantial research effort has been, and will be, devoted toward this desirable goal. Unfortunately, the labels commonly used in the literature to categorize post-injury axonal anatomy might hinder advancement. In this article, we present an argument for the importance of developing precise terms that describe axonal growth in terms of the inciting event, the distance of axonal extension and the timing of axonal growth. The phenotypes produced by molecular interventions that overcome astroglial scar or myelin-associated inhibitors are reframed and discussed in this context. PMID- 18395810 TI - Description and validation of a geriatric multidimensional graphical instrument for promoting longitudinal evaluation. AB - The debate about the adoption of standard multidimensional geriatric assessment instruments is mainly due to the lack of consensus on the feasibility and requirements for such instruments by both the health and the social care professions. A tool called ValGraf was developed in the attempt to give an original answer to these and other controversial issues. The aim of this study was to evaluate the ValGraf for acceptability, concurrent validity and factorial structure. The functional and cognitive impairments as ascertained by the ValGraf were compared with Katz index and Folstein's Mini Mental State Examination (MMSE). Subjects (N=210) of four nursing homes were assessed by two independent teams of nurses. The factorial validity study involved 8280 subjects living in nursing homes. Assessments were conducted throughout the 2001 by 20 geriatricians. The agreement between the ValGraf sections concerning independence in daily living and Katz's index was almost total (r=0.96) and that between ValGraf items on cognition and MMSE was very good (0.73). Factor analysis revealed that 13 coherent factors explained 53% of total variance. ValGraf was proved to be acceptable and comprehensive, criterion valid, at least as daily activities and cognitive status are concerned, and to have a coherent factorial structure. PMID- 18395809 TI - A novel small heat shock protein 12.6 (HSP12.6) from Brugia malayi functions as a human IL-10 receptor binding protein. AB - Phage display cDNA expression library of the third stage larvae (L3) of Brugia malayi was screened for identifying target(s) that bound to the human interleukin 10 receptor (huIL10R). This iterative screening identified an insert that showed significant homology to Caenorhabditis elegans HSP12.6. The gene was designated B. malayi HSP12.6 (BmHSP12.6) and has orthologues in several gastrointestinal nematode genome (Ancylostoma caninum, Ascaris lumbricoides and Ascaris suum) but the gene or gene product has not been studied further in these parasites. Structural analyses of BmHSP12.6 showed that it has a highly conserved alpha crystallin central domain that is characteristic of other small heat shock proteins (HSPs). BmHSP12.6 has a short N-terminal domain and an unusually small C terminal domain flanking the crystallin domain suggesting that this protein belongs to a novel class of small HSPs. BmHSP12.6 appears to be differentially transcribed with highest expression in the vertebrate stages of the parasite (L4, adult and mf) compared to its mosquito vector stage (L3). More importantly recombinant BmHSP12.6 bound to huIL10R in a dose dependent fashion and inhibited the binding of human IL-10 (huIL10) to huIL10R in vitro. rBmHSP12.6 also enhanced the growth and proliferation of MC/9 mast cells in vitro similar to huIL10. This study thus describes a novel small HSP from B. malayi that has the capacity to bind to huIL10R, block binding of huIL10 to huIL10R and function similar to huIL10. PMID- 18395811 TI - Dose-volume and biological-model based comparison between helical tomotherapy and (inverse-planned) IMAT for prostate tumours. AB - BACKGROUND AND PURPOSE: Helical tomotherapy (HT) and intensity-modulated arc therapy (IMAT) are two arc-based approaches to the delivery of intensity modulated radiotherapy (IMRT). Through plan comparisons we have investigated the potential of IMAT, both with constant (conventional or IMAT-C) and variable (non conventional or IMAT-NC, a theoretical exercise) dose-rate, to serve as an alternative to helical tomotherapy. MATERIALS AND METHODS: Six patients with prostate tumours treated by HT with a moderately hypo-fractionated protocol, involving a simultaneous integrated boost, were re-planned as IMAT treatments. A method for IMAT inverse-planning using a commercial module for static IMRT combined with a multi-leaf collimator (MLC) arc-sequencing was developed. IMAT plans were compared to HT plans in terms of dose statistics and radiobiological indices. RESULTS: Concerning the planning target volume (PTV), the mean doses for all PTVs were similar for HT and IMAT-C plans with minimum dose, target coverage, equivalent uniform dose (EUD) and tumour control probability (TCP) values being generally higher for HT; maximum dose and degree of heterogeneity were instead higher for IMAT-C. In relation to organs at risk, mean doses and normal tissue complication probability (NTCP) values were similar between the two modalities, except for the penile bulb where IMAT was significantly better. Re-normalizing all plans to the same rectal toxicity (NTCP=5%), the HT modality yielded higher TCP than IMAT-C but there was no significant difference between HT and IMAT-NC. The integral dose with HT was higher than that for IMAT. CONCLUSIONS: with regards to the plan analysis, the HT is superior to IMAT-C in terms of target coverage and dose homogeneity within the PTV. Introducing dose-rate variation during arc-rotation, not deliverable with current linac technology, the simulations result in comparable plan indices between (IMAT-NC) and HT. PMID- 18395812 TI - New and emerging approaches to understanding developmental coordination disorder. PMID- 18395813 TI - Improved sequence-based prediction of protein secondary structures by combining vacuum-ultraviolet circular dichroism spectroscopy with neural network. AB - Synchrotron-radiation vacuum-ultraviolet circular dichroism (VUVCD) spectroscopy can significantly improve the predictive accuracy of the contents and segment numbers of protein secondary structures by extending the short-wavelength limit of the spectra. In the present study, we combined VUVCD spectra down to 160 nm with neural-network (NN) method to improve the sequence-based prediction of protein secondary structures. The secondary structures of 30 target proteins (test set) were assigned into alpha-helices, beta-strands, and others by the DSSP program based on their X-ray crystal structures. Combining the alpha-helix and beta-strand contents estimated from the VUVCD spectra of the target proteins improved the overall sequence-based predictive accuracy Q(3) for three secondary structure components from 59.5 to 60.7%. Incorporating the position-specific scoring matrix in the NN method improved the predictive accuracy from 70.9 to 72.1% when combining the secondary-structure contents, to 72.5% when combining the numbers of segments, and finally to 74.9% when filtering the VUVCD data. Improvement in the sequence-based prediction of secondary structures was also apparent in two other indices of the overall performance: the correlation coefficient (C) and the segment overlap value (SOV). These results suggest that VUVCD data could enhance the predictive accuracy to over 80% when combined with the currently best sequence-prediction algorithms, greatly expanding the applicability of VUVCD spectroscopy to protein structural biology. PMID- 18395814 TI - An identification of stem cell-resembling gene expression profiles in high-grade astrocytomas. AB - High-grade astrocytomas are among the most intractable types of cancers and are often fatal. Previous studies have suggested that high-grade astrocytomas may adopt the self-renewal and migration properties of neural stem cells (NSCs) to proliferate and spread by expressing the stem cell-specific genes. However, despite a few common molecules being documented, the molecular basis underlying these similarities remains largely unknown. To have a better understanding of the stem cell characteristics of high-grade astrocytomas, we performed the study to identify the stem cell-resembling gene expression profile in high-grade astrocytomas. cDNA microarray analysis was used to detect the differentially expressed genes of isolated human high-grade astrocytomas versus their peritumoral tissue counterparts, and the identification of stem cell-resembling genes was approached by comparing the high-grade astrocytomas-specific gene expression profile with that of NSCs identified by our previous study and other groups. We identified more than 200 high-grade astrocytomas-specific genes in this study, and near 10% genes or gene families of them exhibited similar up or down expression patterns as in NSCs. Further analysis indicated that these genes were actively involved in cell proliferation, adhesion, migration, and metastasis. This study revealed a list of stem cell-specific genes in high-grade astrocytomas, which was likely to have critical roles in determining the "stem" characteristics of high-grade astrocytomas. PMID- 18395815 TI - CLUMPHAP: a simple tool for performing haplotype-based association analysis. AB - The completion of the HapMap Project and the development of high-throughput single nucleotide polymorphism genotyping technologies have greatly enhanced the prospects of identifying and characterizing the genetic variants that influence complex traits. In principle, association analysis of haplotypes rather than single nucleotide polymorphisms may better capture an underlying causal variant, but the multiple haplotypes can lead to reduced statistical power due to the testing of (and need to correct for) a large number of haplotypes. This paper presents a novel method based on clustering similar haplotypes to address this issue. The method, implemented in the CLUMPHAP program, is an extension of the CLUMP program designed for the analysis of multi-allelic markers (Sham and Curtis [1995] Ann. Hum. Genet. 59(Pt1):97-105). CLUMPHAP performs a hierarchical clustering of the haplotypes and then computes the chi(2) statistic between each haplotype cluster and disease; the statistical significance of the largest of the chi(2) statistics is obtained by permutation testing. A significant result suggests that the presence of a disease-causing variant in the haplotype cluster is over-represented in cases. Using simulation studies, we have compared CLUMPHAP and more widely used approaches in terms of their statistical power to identify an untyped susceptibility locus. Our results show that CLUMPHAP tends to have greater power than the omnibus haplotype test and is comparable in power to multiple regression locus-coding approaches. PMID- 18395817 TI - Digital differential interference contrast autofocus for high-resolution oil immersion microscopy. AB - Continued advances in cellular fluorescent biosensors enable studying intracellular protein dynamics in individual, living cells. Autofocus is valuable in such studies to compensate for temperature drift, uneven substrate over multiple fields of view, and cell growth during long-term high-resolution time lapse studies of hours to days. Observing cellular dynamics with the highest possible resolution and sensitivity motivates the use of high numerical aperture (NA) oil-immersion objectives, and control of fluorescence exposure to minimize phototoxicity. To limit phototoxicity, to maximize light throughput of the objective for biosensor studies, and because phase contrast is distorted by the meniscus in microtiter plates, we studied autofocus in differential interference contrast (DIC) microscopy with a 60x 1.45 NA oil objective after removing the analyzer from the fluorescent light path. Based on a study of the experimental DIC modulation transfer function, we designed a new bandpass digital filter for measuring image sharpness. Repeated tests of DIC autofocus with this digital filter on 225 fields-of-view resulted in a precision of 8.6 nm (standard deviation). Autofocus trials on specimens with thicknesses from 9.47 to 33.20 mum, controlled by cell plating density, showed that autofocus precision was independent of specimen thickness. The results demonstrated that the selected spatial frequencies enabled very high-precision autofocus for high NA DIC automated microscopy, thereby potentially removing the problems of meniscus distortion in phase contrast imaging of microtiter plates and rendering the toxicity of additional fluorescence exposure unnecessary. PMID- 18395818 TI - ERp29, an endoplasmic reticulum secretion factor is involved in the growth of breast tumor xenografts. AB - Cancer cells are committed to an actively secretory state that facilitates communication with their microenvironment. We have addressed the role of ERp29, a novel endoplasmic reticulum secretion factor in mammary carcinogenesis using MCF 7 human breast cancer cells as a model. Xenografts originating from cells stably transfected with dominant-negative form of ERp29 were smaller and better differentiated than those derived from cells overexpressing wild-type ERp29. Similar effects were observed by siRNA-mediated ERp29 silencing in xenografts. However, unlike xenografts, the modulation of ERp29 in vitro did not affect the rate of cell proliferation. In addition, we have evaluated the expression of ERp29 in the resting and lactating mammary glands of mice as well as in the human primary breast tumors. About 25% of breast cancers and also lactating mammary glands were expressing ERp29 while the resting glands did not. Taken together these data suggest the active involvement of ERp29 in the malignant conversion of mammary epithelial cells. PMID- 18395819 TI - No pathogenic rearrangement within the DISC 1 gene in psychosis. AB - A translocation disrupting the DISC 1 gene segregates with schizophrenia and related psychiatric disorders in a large Scottish family. Mutation screening of this gene by routine PCR-based methods has remained largely negative. We sought to detect rearrangements affecting DISC 1 in 347 individuals meeting the DSM3R criteria for schizophrenia or schizoaffective disorder, 70 subjects with bipolar disorder and 377 psychiatrically healthy controls, but failed to detect any pathological rearrangement. PMID- 18395820 TI - Electrospinning fabrication of high-trackable catheter tip with gradually graded or gradient flexibility. AB - Increasing demand for a flexible catheter tip exists in the rapidly growing fields of vascular and interventional radiology. This article describes a fabrication technology producing a catheter tip with gradually graded or gradient flexibility. Based on deflection mechanics of a tubular construct, three models were incorporated into structural designs. The models included graded wall thickness, tapered shape, or graded Young's moduli. Electrospinning using elastomeric polymer [poly(L-lactide-co-epsilon-caprolactone), polyurethane] solutions on a transversely and rotationally moving mandrel enabled preparation of three prototype tubular tips by either regio-specific control of wall thickness, tapered mandrel, or regiospecific deposition of polyurethanes with different Young's moduli. All prototype tips exhibited high flexibility at tip ends, as verified using a bifurcated vessel model. PMID- 18395821 TI - Mechanical and flow characterization of Sponceram carriers: Evaluation by homogenization theory and experimental validation. AB - The experimental evidence of the dependence of cell proliferation and differentiation in vitro on the mechanical environment aims to the need of characterization of porous scaffolds in terms of mechanical and flow properties. In this sense, the Young's modulus and intrinsic permeability for three types of Sponceram(R) cell carriers developed for in-vitro applications are here analyzed. Young's modulus and ultimate compression stress were obtained by performing a two plates compression test carried out in a universal microtester machine Instron(R) for several representative samples of each specimen. A permeability test was also implemented to correlate flow rate and pressure gradient in the linear range. Furthermore, porosity and specific surface were obtained through micro-CTs of the scaffold microstructure. These experimental data were compared with those obtained numerically by homogenization for several representative volume elements (RVEs) of the scaffolds microstructure. The good agreement found between numerical and experimental results let us consider that the use of numerical techniques is an attractive tool for the analysis of complex scaffold microstructures. Moreover, Sponceram(R) carriers are shown to have very appropriate properties as bone bioscaffolds which let us recommending further clinical and numerical research on these specific materials. PMID- 18395822 TI - Antimicrobial properties and elution kinetics of linezolid-loaded hip spacers in vitro. AB - Bacterial adhesion to and -persistence on antibiotic-loaded bone cement is an increasing problem. New antibiotics with good antimicrobial and pharmacokinetic properties (e.g. linezolid) may be the solution to this problem; however, few data concerning linezolid-loaded acrylic cement are currently available. Ten gentamicin-linezolid-loaded hip spacers (1 g gentamicin/2.4 g linezolid/80 g PMMA; five spacers including a metallic endoskeleton, five with no metallic components) were tested in vitro against a strain of methicillin-resistant Staphylococcus aureus with regard to antibiotic release and bacteria growth inhibition. Daily, the antibiotic elution was determined by high liquid performance chromatography (linezolid) and fluorescence polarization immunoassay (gentamicin), the bacteria growth inhibition photometrically at 546 nm. All spacers demonstrated growth inhibition for 8 days. Peak average concentrations were reached for both agents on day 1 (gentamicin-35.10 mug/mL [24.10-52.52], linezolid-36.28 mug/mL [22.87-71.76]). After 8 days, 0.97% [0.93-1.05%] of the initial amount of linezolid and 3.13% [2.85-3.31%] of gentamicin were meanly released from spacers containing a metallic endoskeleton. In those containing of simple cement these values were 1.22% [0.91-1.59%] and 2.67% [2.12-2.73%], respectively. Linezolid demonstrated acceptable elution kinetics from bone cement; however, further experimental research and animal studies should clarify any possible side effect of linezolid-loaded cement media before definitive use in the clinical practice. PMID- 18395823 TI - Early healing of nanothickness bioceramic coatings on dental implants. An experimental study in dogs. AB - Thick bioceramic coatings like plasma sprayed hydroxyapatite have been shown to increase the overall tissue response and biomechanical fixation of dental implants. However, the presence and potential fracture of a bone-coating-metallic substrate interface at long times after implantation led these implants to fall from favor in clinical practice. The purpose of this study was to evaluate the biomechanical fixation and biological response of Ca- and P-based, 20-50 nm thickness bioceramic deposition on a previously alumina-blasted/acid-etched Ti 6Al-4V implant surface in a dog model. Cylindrical alumina-blasted/acid-etched (AB/AE) (Control, n = 16), and Nanothickness bioceramic coated AB/AE(Nano, n = 16) implant surfaces were surgically placed in dogs proximal tibia and remained for 2 and 4 weeks in vivo. Following euthanization, the implants-in-bone were mounted in epoxy and pullout at a 0.5 mm/min rate. Following mechanical testing, the specimens were decalcified and processed (Hematoxylin and Eosin) for standard transmitted light microscopy evaluation. Percent bone-to-implant contact (BIC) to the pulled out implant surface was determined through computer software. Statistical analyses were performed by one-way ANOVA at 95% level of significance and Tukey's post-hoc multiple comparisons. No significant differences in pullout force were observed (p > 0.88): 2W Control (212.08 +/- 42.96 N), 2W Nano (224.35 +/- 42.97 N), 4W Control (207.07 +/- 42.97 N), and 4W Nano (190.15 +/- 45.94 N). No significant differences in %BIC were observed (p > 0.94): 2W Control (72.66 +/ 8.51), 2W Nano (69.44 +/- 8.51), 4W Control (70.44 +/- 8.51), and 4W Nano (69.11 +/- 9.09). It is shown that 20-50 nm thickness bioceramic depositions onto previously alumina-blasted/acid-etched substrates did not improve the biomechanical fixation and the BIC at early implantation times, and studies concerning shorter and longer implantation times are recommended for confirmation or before a conclusion can be made. PMID- 18395824 TI - Effects of monomers eluted from dental resin restoratives on osteoblast-like cells. AB - Resin-modified glass-ionomer (RMGIC) has been demonstrated to exhibit inhibition on the growth and differentiation of osteoblasts on its surface. In this study, the hypothesis that the different levels of inhibitory effects on osteoblasts of resin restoratives depend on elution of unpolymerized monomers was examined. Release of monomers from cured specimens of Bis-GMA/TEGDMA-composites, MMA-resin cements, or HEMA-containing RMGIC was determined and osteoblast-like MC3T3-E1 cells were cultured in the presence of 100-10 microg/mL of TEGDMA, 10-1 microg/mL of MMA, or 400-50 microg/mL of HEMA according to the release concentrations. Cell proliferation, alkaline phosphatase (ALP) activity, the expression of osteoblastic markers, and mineralized tissue formation were evaluated. TEGDMA and MMA at the concentrations tested did not affect the growth of MC3T3-E1 and exhibited little harmful effects on their differentiation and mineralization. On the contrary, HEMA inhibited proliferation, ALP activities, the expression of osteocalcin, and mineralized tissue formation at 200 microg/mL or more. These results indicate that HEMA at the concentrations similar to that observed in elution tests affected osteoblastic proliferation, differentiation, and mineralization, suggesting that elution of unreacted HEMA could be the main component of the adverse effects of RMGIC on osteoblast-like cells and influences of resin restoratives on the osteoblasts are possibly dependant on release characteristics of unpolymerized monomers. PMID- 18395825 TI - Three-layered membranes of collagen/hydroxyapatite and chitosan for guided bone regeneration. AB - The purpose of this study was to design a novel hybrid membrane with optimized properties for guided bone regeneration (GBR). Both the top and bottom layers of the sandwich-structured membrane were composed of collagen containing 20 wt% hydroxyapatite (HA), while the middle layer was made of chitosan. The above three layers were formulated into an integral membrane from their respective slurries through a layer-by-layer filtration process. The phase and composition of the membrane were confirmed by FT-IR and XRD analyses. The observation of its morphology by SEM showed that the membrane had a porous structure and structural integrity. The chitosan layer ensured the high tensile strength and elastic modulus of the membrane, while the presence of the collagen/HA composite layers endowed it with good flexibility and bioactivity. These results suggest that the integrated membrane prepared in this study would have the potential for use as a GBR material. PMID- 18395827 TI - Neuronal assemblies of the Drosophila mushroom body. AB - The mushroom body (MB) of the insect brain has important roles in odor learning and memory and in diverse other brain functions. To elucidate the anatomical basis underlying its function, we studied how the MB of Drosophila is organized by its intrinsic and extrinsic neurons. We screened for the GAL4 enhancer-trap strains that label specific subsets of these neurons and identified seven subtypes of Kenyon cells and three other intrinsic neuron types. Laminar organization of the Kenyon cell axons divides the pedunculus into at least five concentric strata. The alpha', beta', alpha, and beta lobes are each divided into three strata, whereas the gamma lobe appears more homogeneous. The outermost stratum of the alpha/beta lobes is specifically connected with a small, protruded subregion of the calyx, the accessory calyx, which does not receive direct olfactory input. As for the MB extrinsic neurons (MBENs), we found three types of antennal lobe projection neurons, among which two are novel. In addition, we resolved 17 other types of MBENs that arborize in the calyx, lobes, and pedunculus. Lobe-associated MBENs arborize in only specific areas of the lobes, being restricted along their longitudinal axes, forming two to five segmented zones in each lobe. The laminar arrangement of the Kenyon cell axons and segmented organization of the MBENs together divide the lobes into smaller synaptic units, possibly facilitating characteristic interaction between intrinsic and extrinsic neurons in each unit for different functional activities along the longitudinal lobe axes and between lobes. Structural differences between lobes are also discussed. PMID- 18395826 TI - Enzymatic biodegradation of HEMA/bisGMA adhesives formulated with different water content. AB - Dentin adhesives may undergo phase separation when bonding to wet demineralized dentin. We hypothesized that adhesives exhibiting phase separation will experience enhanced biodegradation of methacrylate ester groups. The objective of this project was to study the effect of enzyme-exposure on the release of methacrylic acid (MAA) and 2-hydroxyethyl methacrylate (HEMA) from adhesives formulated under conditions simulating wet bonding. HEMA/bisGMA(2,2-bis[4(2 hydroxy-3-methacryloyloxy-propyloxy)-phenyl] propane), 45/55 w/w ratio, was formulated with different water content: 0 Wt % (A00), 8 wt % (A08), and 16 wt % (A16). After a three day prewash, adhesive discs were incubated with/without porcine liver esterase (PLE) in phosphate buffer (PB, pH 7.4) at 37 degrees C for 8 days. Supernatants were collected daily and analyzed for MAA and HEMA by HPLC. For all formulations, daily MAA release in the presence of PLE was increased compared to MAA release in PB. HEMA release in the presence of PLE was not detected while HEMA release was consistently measured in PB. A08 and A16 released significantly larger amounts of HEMA compared to A00. Analysis of the cumulative release of analytes showed that the leachables in PLE was significantly increased (p < 0.05) as compared with that released in PB indicating that MAA release was not only formed from unreacted monomers but from pendant groups in the polymer network. However, the levels of analytes HEMA in PB or MAA in PLE were increased in A08 and A16 as compared with A00, which suggests that there could be a greater loss of material in HEMA/bisGMA adhesives that experience phase separation under wet bonding conditions. PMID- 18395828 TI - Thalamo-telencephalic pathways in the fire-bellied toad Bombina orientalis. AB - It was suggested that among extant vertebrates, anuran amphibians display a brain organization closest to the ancestral tetrapod condition, and recent research suggests that anuran brains share important similarities with the brains of amniotes. The thalamus is the major source of sensory input to the telencephalon in both amphibians and amniote vertebrates, and this sensory input is critical for higher brain functions. The present study investigated the thalamo telencephalic pathways in the fire-bellied toad Bombina orientalis, a basal anuran, by using a combination of retrograde tract tracing and intracellular injections with the tracer biocytin. Intracellular labeling revealed that the majority of neurons in the anterior and central thalamic nuclei project to multiple brain targets involved in behavioral modulation either through axon collaterals or en passant varicosities. Single anterior thalamic neurons target multiple regions in the forebrain and midbrain. Of note, these neurons display abundant projections to the medial amygdala and a variety of pallial areas, predominantly the anterior medial pallium. In Bombina, telencephalic projections of central thalamic neurons are restricted to the dorsal striato-pallidum. The bed nucleus of the pallial commissure/thalamic eminence similarly targets multiple brain regions including the ventral medial pallium, but this is accomplished through a higher variety of distinct neuron types. We propose that the amphibian diencephalon exerts widespread influence in brain regions involved in behavioral modulation and that a single dorsal thalamic neuron is in a position to integrate different sensory channels and distribute the resulting information to multiple brain regions. PMID- 18395829 TI - The use of nanoindentation for characterizing the properties of mineralized hard tissues: state-of-the art review. AB - The use of nanoindentation to determine nanomechanical properties of mineralized tissues has been investigated extensively. A detailed, critical, and comprehensive review of this literature is the subject of the present work. After stating the motivation for the review, a succinct presentation of the challenges, advantages, and disadvantages of the various quasi-static nanoindentation test methods (to obtain elastic modulus, E, and hardness, H) and dynamic test methods (to obtain storage and loss moduli and/or loss/damping factor) is given in the form of a primer. Explicative summaries of literature reports on various intrinsic and extrinsic factors that significantly influence E and H, followed by 15 suggested topics for future research, are included additionally. This review is designed to present a compact guide to the principles of the nanoindentation technique and to emphasize considerations when determining material properties of mineralized tissues. PMID- 18395830 TI - Function of Pax1 and Pax9 in the sclerotome of medaka fish. AB - We examined the expression and functions of Pax1 and Pax9 in a teleost fish, the medaka Oryzias latipes. While Pax1 and Pax9 show distinct expression in the sclerotome in amniotes, we could not detect the differential expression of Pax1 and Pax9 in the developing sclerotome of the medaka. Furthermore, unlike the mouse, in which Pax1 is essential for development of the vertebral body, and where the neural arch is formed independent of either Pax1 or Pax9, our morpholino knockdown experiments revealed that both Pax1 and Pax9 are indispensable for the development of the vertebral body and neural arch. Therefore, we conclude that after gene duplication, Pax1 and Pax9 subfunctionalize their roles in the sclerotome independently in teleosts and amniotes. In Stage-30 embryo, Pax9 was strongly expressed in the posterior mesoderm, as was also observed for mouse Pax9. Since this expression was not detected for Pax1 in the mouse or fish, this new expression in the posterior mesoderm likely evolved in Pax9 of ancestral vertebrates after gene duplication. Two-month-old fish injected with Pax9 morpholino oligonucleotide showed abnormal morphology in the tail hypural skeletal element, which may have been related to this expression. PMID- 18395831 TI - Induction of apoptosis in SF21 cell line by conditioned medium of the entomopathogenic fungus, Nomuraea rileyi, through Sf-caspase-1 signaling pathway. AB - The apoptosis in SF-21 cell line can be induced by the conditioned medium (CM) of the entomopathogenic fungus, Nomuraea rileyi, based on changes in morphology and formation of apoptotic bodies in cultured cells, and with the onset of DNA fragmentation as shown by TUNEL staining and agarose electrophoresis. Moreover, the induction of apoptosis in SF-21 cells was inhibited by adding the inhibitor of effector caspase, viz. z-DEVD-fmk, to the CM, indicating that Sf-caspase-1 is involved in this apoptosis. Similarly, the inhibitor of initiator caspase, viz., z-VAD-fmk, inhibited apoptosis. Therefore, both initiator and effector caspases are possibly involved in the apoptosis of SF-21 cells. In addition, we detected Sf-caspase-1 activity in the process of apoptosis in SF-21 cells, suggesting that the effector caspase in SF-21 is similar to that found in mammalian cells. Our results also indicated that the apoptosis found in this line is accomplished through a Sf-caspase-1 signaling pathway. PMID- 18395832 TI - Reduction of phosphorylated histone H3 serine 10 and serine 28 cell cycle marker intensities after DNA damage. AB - Phosphorylated histone H3 at serine 10 and serine 28 (H3Ser10 and H3Ser28) have been recognized as cell cycle markers to evaluate the late-G(2)/M status of cell and tissue samples. I report about the reduction phenomena of H3Ser10 and Ser28 phosphorylation (H3Ser10P and 28P) at the late-G(2) through M cell cycle phases in association with DNA damage caused by hydrogen peroxide (H(2)O(2)). The levels of H3Ser10P and Ser28P decreased between 15 and 60 min after H(2)O(2) addition in an inverse correlation manner with H2AX Ser139 phosphorylation (gammaH2AX). Experiments using wortmannin suggested that the reduction events of H3Ser10P/28P are under the control of phosphatidylinositol 3-kinase-like kinases. Fluorescence microscopic observation showed that H3Ser10 and Ser28 on telomeric portions of condensed M-phase chromosomes retained their strongly phosphorylated status even after 60 min of H(2)O(2) treatment. In addition, these chromosome parts were poorly gammaH2AX positive showing mutually exclusive distribution patterns between H3Ser10P/28P and gammaH2AX. Considering these data, I hypothesize that the reduction of the H3Ser10P/28P is a part of the DNA damage response processes. It is advisable to pay careful attention to these phenomena at the time of designing cell cycle assay protocols with H3Ser10P or Ser28P mitosis markers when DNA damaging process is expected to occur. PMID- 18395833 TI - Potential involvement of the interaction between insulin-like growth factor binding protein (IGFBP)-6 and LIM mineralization protein (LMP)-1 in regulating osteoblast differentiation. AB - Insulin-like growth factor binding protein (IGFBP)-6 has been reported to inhibit differentiation of myoblasts and osteoblasts. In the current study, we explored the mechanisms underlying IGFBP-6 effects on osteoblast differentiation. During MC3T3-E1 osteoblast differentiation, we found that IGFBP-6 protein was down regulated. Overexpression of IGFBP-6 in MC3T3-E1 and human bone cells inhibited nodule formation, osteocalcin mRNA expression and ALP activity. Furthermore, accumulation of IGFBP-6 in the culture media was not required for any of these effects suggesting that IGFBP-6 suppressed osteoblast differentiation by an intracellular mechanism. A yeast two-hybrid screen of an osteosarcoma library was conducted to identify intracellular binding partners to account for IGFBP-6 inhibitory effects on osteoblast differentiation. LIM mineralizing protein (LMP 1) was identified as a high affinity IGFBP-6 binding partner. Physical interaction between IGFBP-6 and LMP-1 was confirmed by co-immunoprecipitation. Fluorescent protein fusion constructs for LMP-1 and IGFBP-6 were transiently transfected into osteoblasts to provide evidence of subcellular locations for each protein. Coexpression of LMP-1-GFP and IGFBP-6-RFP resulted in overlapping subcellular localization of LMP-1 and IGFBP-6. To determine if there was a functional association of IGFBP-6 and LMP-1 as well as a physical association, we studied the effect of IGFBP-6, LMP-1 and their combination on type I procollagen promoter activity. LMP-1 increased promoter activity while IGFBP-6 reduced promoter activity, and coexpression of LMP-1 with IGFBP-6 abrogated IGFBP-6 suppression. These studies provide evidence that overexpression of IGFBP-6 suppresses human and murine osteoblast differentiation, that IGFBP-6 and LMP-1 physically interact, and supports the conclusion that this interaction may be functionally relevant. PMID- 18395834 TI - Use of ERT2-iCre-ERT2 for conditional transgenesis. AB - We examined the use of ERT2-iCre-ERT2 (Cre2ERT2), a tamoxifen-regulated form of Cre that has been described to have a background activity lower than that of other tamoxifen-regulated Cre constructs, for establishing performant conditional deleter mouse lines. Cre2ERT2 was inserted by homologous recombination into the Rosa26 locus. These mice were mated with R26R Cre-reporter mice. No recombination could be observed in the progenies in the absence of tamoxifen treatment. Tamoxifen treatment at E13-14 led to a high level, albeit variable, recombination in most of the tissues examined: liver, heart, kidney, brain, lung etc. Treatment of adult animals also induced recombination in these tissues, although at a lower level. Northern blot and qPCR studies suggested that these differences are not linked to significant variations of the level of expression of Cre2ERT2. Thus, Cre2ERT2 appears to be a good alternative to existing modulatable Cre systems, displaying a lack of background activity and a high-level inducibility in vivo. PMID- 18395835 TI - Cre reporter mouse expressing a nuclear localized fusion of GFP and beta galactosidase reveals new derivatives of Pax3-expressing precursors. AB - A new Cre-reporter strain of mouse has been developed that expresses a fusion protein derived from the lacZ gene fused to GFP with a nuclear localization signal. This construct is expressed from the ROSA26 locus upon Cre-mediated recombination that removes a loxP-flanked PGK-neo cassette, thus allowing for detection of Cre activity in all tissues. This reporter strain, which is similar to prior R26R and R26EGFP strains, has certain advantages related to the nuclear expression and the combined expression of both beta-galactosidase and GFP activities. We show that the use of this newly developed reporter line allows for enhanced resolution, detection and co-localization. Thus, we report a previously unrecognized subset of venous endothelial cells derived from Pax3 expressing precursors. PMID- 18395836 TI - The localization of PMCA1b in epithelial cells and aposomes of the rat coagulating gland is influenced by androgens. AB - BACKGROUND: Rat coagulating gland epithelial cells export proteins by an apocrine secretion mode within membrane blebs arising from the apical plasma membrane. Using a pan-PMCA antibody, we have recently shown the plasma membrane Ca(2+) ATPase (PMCA) being part of the apical plasma membrane of epithelial cells and incorporated into the aposomal membrane. The mRNA of PMCA isoforms 1 and 4 respectively, have been detected by RT-PCR in rat coagulating gland. METHODS: In order to identify which PMCA isoform is integrated into aposomes during apocrine secretion and whether or not PMCA export is influenced by androgens RT-PCR, in situ hybridization, Western blotting, and immunofluorescence experiments were performed. RESULTS: PMCA1b is the isoform which is expressed and located in the apical plasma membrane of coagulating gland epithelial cells and is integrated into the aposomal membrane. In contrast, PMCA4 mRNA and protein are restricted to the stroma. Androgen deprivation by castration within 14 days leads to an accumulation of PMCA1b in coagulating gland epithelium, while aposomes are not detected anymore. CONCLUSIONS: We showed for the first time that PMCA isoform 1b is released via aposomes of the epithelial cells of the rat coagulating gland and that the localization of PMCA1b in the epithelial cells is influenced by androgens. PMID- 18395837 TI - Mouse Dach1 and Dach2 are redundantly required for Mullerian duct development. AB - dachshund/Dach gene family members encode transcriptional cofactors with highly conserved protein interaction domains and are expressed in the developing eyes, brains, and limbs in insects and vertebrates. These observations suggest that the developmental roles of dachshund/Dach in these tissues have been conserved since the divergence of arthropods and chordates. However, while Drosophila dachshund mutants have abnormalities in eye, brain, limbs, mouse Dach1 or Dach2 knockout mutants do not exhibit gross anatomical malformations in these tissues. In addition, Dach1/2 double homozygotes have intact eyes and limbs. Here we show that in Dach1/Dach2 double mutants, female reproductive tract (FRT) development is severely disrupted. This defect is associated with the Mullerian duct (MD) and not the Wolffian duct (WD), which normally differentiate into either the FRT or male reproductive tract (MRT), respectively. Dach1 and Dach2 are expressed in the MD, and in Dach1/2 double mutants, MD expression of Lim1 and Wnt7a is abnormal and MD development is disrupted. In contrast, WD and MRT development are not grossly affected. We propose that Dach1 and Dach2 proteins may redundantly control FRT formation by regulating the expression of target genes required for development of the MD. This vertebrate Dach1/2 function may have been conserved during arthropod evolution, as Drosophila dachshund mutants also exhibit an FRT phenotype. PMID- 18395839 TI - Temporally controlled prostate epithelium-specific gene alterations. AB - Employing the Hprt locus as the site for targeted transgenesis we have developed mice expressing the tamoxifen-inducible Cre-ER(T2) fusion protein under the control of the ARR2-rat probasin promoter. This system enables external control over the timing of prostate epithelial cell-specific gene alterations. Using both the ROSA26-lacZ and ROSA26-EYFP reporter strains to monitor recombinase activity, Cre-ER(T2) was found to be specifically expressed in the prostatic epithelium and was strictly tamoxifen dependent. This strain thus allows precise control over the timing of gene alterations in the mouse prostate, enabling analyses of the phenotypic consequences of gene alterations in mice of any age. It also provides an ideal platform to study the impact of environmental, hormonal, and age-related factors on prostate tumorigenesis. This latter feature will be of particular value given the paucity of murine models that accurately mimic the late onset and prolonged natural history of human prostate cancer. PMID- 18395840 TI - Developmental toxicity assessment of thermoresponsive poly(N-isopropylacrylamide co-acrylamide) oligomers in CD-1 mice. AB - BACKGROUND: Although polymers and hydrogels are used successfully in biomedical applications, including implants and drug delivery devices, smaller molecular weight oligomers, such as those investigated here, have not been extensively studied in vivo. Poly(N-isopropylacrylamide-co-acrylamide), or P(NIPAAm-co-AAm), has a unique thermoresponsive behavior and is under investigation as a novel drug delivery system for metastatic cancer treatment. To date, no studies have been published regarding the safety of P(NIPAAm-co-AAm) to the conceptus. METHODS: From gestation days (GD) 6-16, pregnant CD-1 mice were dosed via i.p. injection with aqueous solutions containing 500, 750, or 1,000 mg/kg/d P(NIPAAm-co-AAm). Dams were sacrificed on GD 17 and their litters were examined for abnormalities. RESULTS: P(NIPAAm-co-AAm) caused no statistically significant difference in maternal weight gain or percent resorbed or dead fetuses compared to control values, but fetal weight was significantly decreased in the two highest dosage groups. CONCLUSIONS: At the highest dosages employed, maternal exposure to P(NIPAAm-co-AAm) was associated with decreased fetal weight. However, as the estimated human exposure levels for persons using this system would be some 1,500 fold lower than the lowest dosage administered in this study, the authors feel that this oligomer was not shown to pose a biologically significant risk at relevant human dosages. PMID- 18395841 TI - Selective portal vein embolization and ligation trigger different regenerative responses in the rat liver. AB - Two strategies are clinically available to induce selective hypertrophy of the liver: portal vein embolization (PVE) and portal vein ligation (PVL). The aim of this study was to compare the impact of PVE and PVL on liver regeneration. Rats were subjected to 70% PVL, 70% PVE, 70% partial hepatectomy (PH) (positive control), or sham operation (negative control). PVL and PVE of liver segments were validated by portography and histology, demonstrating obstruction of the involved portal branches. Liver weight and markers of regeneration were assessed at 24, 48, and 72 hours, and 7 days after surgery (n = 5). Sinusoidal perfusion was examined by intravital microscopy. The weight of the regenerating liver segments increased continuously in all groups, with the highest weight gain after PH, which also disclosed the strongest proliferative activity. In Ki-67 and PCNA stainings, hepatocyte proliferation after PVL was more pronounced than after PVE (P = 0.01). Volumetric blood flow and functional sinusoidal density were lower after PVE than after PVL (P = 0.006, P = 0.02, respectively). The accumulation of Kupffer cells 24 hours after the intervention was highest after PH. Transcript levels of cytokines (interleukin-1beta, tumor necrosis factor-alpha, interleukin 6) peaked at 24 hours and were highest after PH. The embolized part of the liver after PVE showed prominent foreign body reaction in the portal triad with accumulation of macrophages. CONCLUSION: PVL is superior to PVE in inducing a regenerative response of the remnant liver. The impairment of liver regeneration after PVE may be a consequence of macrophage trapping in the occluded segment due to a foreign body reaction. Lower blood flow and lower accumulation of macrophages, particularly Kupffer cells, in the regenerating part of the liver likewise causes impaired liver regeneration after PVE. PMID- 18395838 TI - The genetics of essential metal homeostasis during development. AB - The essential metals copper, zinc, and iron play key roles in embryonic, fetal, and postnatal development in higher eukaryotes. Recent advances in our understanding of the molecules involved in the intricate control of the homeostasis of these metals and the availability of natural mutations and targeted mutations in many of the genes involved have allowed for elucidation of the diverse roles of these metals during development. Evidence suggests that the ability of the embryo to control the homeostasis of these metals becomes essential at the blastocyst stage and during early morphogenesis. However, these metals play unique roles throughout development and exert pleiotropic, metal specific, and often cell-specific effects on morphogenesis, growth, and differentiation. Herein, we briefly review the major players known to be involved in the homeostasis of each of these essential metals and their known roles in development. PMID- 18395842 TI - Lymphoid neogenesis and immune infiltration in aged liver. AB - Immune dysregulation and inflammation play a major role in the pathology of age related disorders. In an earlier study, the microarray data from our laboratory indicated an increase in inflammation-related gene expression in the liver with age. We further investigated immune-related changes in the aged liver and found that the levels of inflammatory cytokines, chemokines, and inflammatory genes were higher in aged animals. Immunohistochemical studies showed that immune cells formed clusters or foci in the livers of old mice, preferentially near the perivascular regions. Further analysis revealed an enrichment of macrophages, T cells, B cells, natural killer cells, and neutrophils in old liver. Characterization of the immune clusters showed the presence of shared markers of tertiary lymphoid neogenesis. Levels of lymph node homing cytokines were elevated. Expression of immunoglobulin and recombinase gene transcripts was also higher, indicating the presence of ectopic lymphoid structures in the aged liver. CONCLUSION: Aged liver exhibits a marked inflammatory status accompanied by increased immune cell infiltration. Inflammation and ectopic lymphoid structures have previously been shown to be associated with carcinogenesis, a condition that becomes more prevalent with age. Thus, further study of inflammation-related changes in the microenvironment of the aged liver could provide insights into these disorders. PMID- 18395843 TI - Mesenchymal stem cell-derived molecules directly modulate hepatocellular death and regeneration in vitro and in vivo. AB - Orthotopic liver transplantation is the only proven effective treatment for fulminant hepatic failure (FHF), but its use is limited because of organ donor shortage, associated high costs, and the requirement for lifelong immunosuppression. FHF is usually accompanied by massive hepatocellular death with compensatory liver regeneration that fails to meet the cellular losses. Therefore, therapy aimed at inhibiting cell death and stimulating endogenous repair pathways could offer major benefits in the treatment of FHF. Recent studies have demonstrated that mesenchymal stem cell (MSC) therapy can prevent parenchymal cell loss and promote tissue repair in models of myocardial infarction, acute kidney failure, and stroke through the action of trophic secreted molecules. In this study, we investigated whether MSC therapy can protect the acutely injured liver and stimulate regeneration. In a D galactosamine-induced rat model of acute liver injury, we show that systemic infusion of MSC-conditioned medium (MSC-CM) provides a significant survival benefit and prevents the release of liver injury biomarkers. Furthermore, MSC-CM therapy resulted in a 90% reduction of apoptotic hepatocellular death and a three fold increment in the number of proliferating hepatocytes. This was accompanied by a dramatic increase in the expression levels of 10 genes known to be up regulated during hepatocyte replication. Direct antiapoptotic and promitotic effects of MSC-CM on hepatocytes were demonstrated using in vitro assays. CONCLUSION: These data provide the first clear evidence that MSC-CM therapy provides trophic support to the injured liver by inhibiting hepatocellular death and stimulating regeneration, potentially creating new avenues for the treatment of FHF. PMID- 18395844 TI - Chronic genotype 4 hepatitis C. PMID- 18395845 TI - Mitochondrial dysfunction by complex II inhibition delays overall cell cycle progression via reactive oxygen species production. AB - Mitochondrial complex II defect has recently been implicated in cellular senescence and in the ageing process of which a critical phenotype is retardation and arrest of cellular growth. However, the underlying mechanisms of how complex II defect affects cellular growth, remain unclear. In this study, we investigated the effect of complex II inhibition using a subcytotoxic dose (400 microM) of 2 thenoyltrifluoroacetone (TTFA), a conventional complex II inhibitor, on cell cycle progression. TTFA (400 microM) directly decreased KCN-sensitive cellular respiration rate to 67% of control and disrupted the mitochondrial membrane potential. In contrast to other respiratory inhibitors such as rotenone, antimycin A, and oligomycin, TTFA prolonged the duration of each phase of the cell cycle (G1, S, and G2/M) equally, thereby delaying overall cell cycle progression. This delay was accompanied by a biphasic increase of reactive oxygen species (ROS) and concurrent glutathione oxidation, in addition to a slight decrease in the cellular ATP level. Finally, the delay in cell cycle progression caused by TTFA was proved to be mainly due to ROS overproduction and subsequent oxidative stress, as evidenced by its reversal following pretreatment with antioxidants. Taken together, these results suggest that an overall delay in cell cycle progression due to complex II defects may contribute to ageing and degenerative diseases via inhibition of cellular growth and proliferation without arrest at any specific phase of the cell cycle. PMID- 18395846 TI - Quantitative comparison of PTH1R in breast cancer MCF7 and osteosarcoma SaOS-2 cell lines. AB - The aim of the present study was to compare the classical parathyroid hormone/parathyroid hormone-related peptide (PTH/PTHrP) receptors in MCF7 breast cancer cells with SaOS-2 osteosarcoma cell line. Quantitative binding showed that (125)I-PTHrP-1-34(Tyr) binds with a single binding site in both cells. However (125)I-PTHrP-1-34(Tyr) has higher affinity binding in MCF7 (K(D) = 1.88 +/- 0.08 nM) than in SaOS-2 cells (K(D) = 4.4 +/- 0.185 nM). The competitive binding using 3.3 nM (125)I-PTHrP-1-34(Tyr) with increasing amounts (0.33-33 nM) of unlabelled human PTHrP-1-34, PTHrP-7-34, PTHrP-1-86 His(5)-PTHrP-1-36, His(5)-Phe(23)-PTHrP 1-36 or PTH-1-34 revealed different displacements. In SaOS-2 the PTHrP-7-34 and PTHrP-1-86 caused similar displacement compared with 73% by PTH-1-34 and 70% by PTHrP-1-34. However, in MCF7, PTHrP-7-34, PTHrP-1-86 and PTH-1-34 displaced by 54%, 72% and 67%, respectively, compared to 87% by PTHrP-1-34. The His(5)-Phe(23) PTHrP-1-36 caused an increase in the K(D) from 2.0 +/- 0.03 nM to 2.75 +/- 0.045 nM in MCF7 cells, but had no significant effect in SaOS-2 cells. The PTH/PTHrP receptor in both cell lines revealed a single 85 KDa band with different intensity. Our results suggest that the PTH/PTHrP receptor in MCF7 cells has higher binding affinity for PTHrP than PTH compared to the receptor in SaOS-2 cells. PMID- 18395847 TI - Restoring the balance: harnessing regulatory T cells for therapy in rheumatoid arthritis. AB - Treg play a vital role in the maintenance of tolerance to self antigens, thereby preventing disease through the active suppression of proliferation and pro inflammatory cytokine production by autoreactive T cells. Here we discuss strategies aimed at enhancing Treg function in patients with rheumatoid arthritis with the ultimate aim of restoring lasting tolerance but without increasing the risk of infections or cancer. PMID- 18395848 TI - Impaired function of dendritic cells translocating the liver sinusoids: a veto effect contributing to intrahepatic tolerance? AB - Due to its unique architecture and conditions of blood flow, the liver is acknowledged as an immunologically unusual organ associated with primary activation of naive T cells and the induction of tolerance. Several mechanisms have been proposed to be involved in this process. Most suggest that naive T cells activated in situ in the hepatic sinusoids are deleted or silenced following activation by liver cells acting as antigen presenting cells. Hepatocytes, liver sinusoidal endothelial cells and bone marrow-derived cells (including Kupffer cells and DC) have been shown to support primary activation in situ and play some role in tolerance induction. Although most liver DC have been described to be immature and located in sites inaccessible to naive T cells, some blood-borne DC have been shown to translocate via the sinusoids where naive T cells recirculate. Thus, the presence of mature DC with potential immunogenicity in the sinusoids might give contradictory signals to the naive T cells activated within this organ. In this issue of the European Journal of Immunology, liver sinusoidal endothelial cells are shown to impair the DC ability to induce the proliferation of naive T cells in vitro via an unknown mechanism. Although these findings need to be confirmed in a physiological setting, regulation of the function of DC translocating the sinusoids might represent a new mechanism contributing to T cell tolerance in the liver. PMID- 18395849 TI - TLR4 and S1P receptors cooperate to enhance inflammatory cytokine production in human gingival epithelial cells. AB - Toll-like receptors (TLR) are pattern recognition receptors for highly conserved microbial molecular patterns. Activation of TLR is a pivotal step in the initiation of innate, inflammatory, and immune defense mechanisms. Recent findings indicate that G protein-coupled receptors (GPCR) may modulate TLR signaling, but it is unclear which GPCR are involved in this process. One such cooperation between GPCR and TLR can be attributed to the sphingosine 1-phosphate (S1P) receptor family. The S1P receptors (S1P1-5) are a family of GPCR with a high affinity for S1P, a serum-borne bioactive lipid associated with diverse biological activities such as inflammation and healing. In this study, we show that pro-inflammatory cytokine production, including IL-6 and IL-8, was increased with LPS and concomitant S1P stimulation. Furthermore, elevated cytokine production following LPS and S1P challenge in human gingival epithelial cells (HGEC) was significantly reduced when TLR4, S1P1 or S1P3 signaling was blocked. Our study also shows that S1P1 and S1P3 expression was induced by LPS in HGEC, and this elevated expression enhanced the influence of S1P in its cooperation with TLR4 to increase cytokine production. This cooperation between TLR4 and S1P1 or S1P3 demonstrates that TLR4 and GPCR can interact to enhance cytokine production in epithelial cells. PMID- 18395850 TI - N-terminal flanking residues of a diabetes-associated GAD65 determinant are necessary for activation of antigen-specific T cells in diabetes-resistant mice. AB - A diabetes-associated peptide in the glutamic acid decarboxylase 65 (GAD65) molecule, p524-543, activates two distinct populations of T cells, which apparently play opposite roles in the development of diabetes in NOD mice. By comparing the fine specificity of these two T cell repertoires using a nested set of truncated peptides that cover the p524-543 region, we found, surprisingly, that all clones recognized the same core within this peptide, p530-539. The core itself was non-immunogenic, but the residues flanking this shared sequence played the crucial role in selecting T cells to activate. A peptide missing N-terminal flanking residues at position 528 and 529 was stimulatory in NOD but not in MHC matched, NOD-resistant (NOR) mice, suggesting that a protective response in the resistant mice may require T cell recognition of one or more of the N-terminal flanking residues. T cell repertoire studies demonstrated selective clonal expansions within the BV4 TCR family that dominates the p524-543 response in NOD but not in NOR mice. These data suggest that processing or trimming events affecting T cell recognition of very few flanking residues of diabetes-associated determinants might be involved in the protective response in NOR mice. PMID- 18395851 TI - CD137 ligand reverse signaling has multiple functions in human dendritic cells during an adaptive immune response. AB - T cell activation via dendritic cells (DC) is an important step in the adaptive immune response, which requires DC maturation, migration to lymph nodes and presentation of antigen to T cells. CD137 receptor expressed on activated T cells is a potent costimulatory molecule. Here, we investigated the functions of CD137 ligand (CD137L) in human monocyte-derived DC during an immune response. Cross linking of CD137L on DC leads to cell maturation in an autocrine fashion, mostly via release of TNF-alpha. Reverse signaling of CD137L also mediates migration of DC via up-regulation of the CCR7 chemokine receptor, demonstrated by an in vivo MIP-3beta-dependent SCID mouse migration model. Finally, CD137L-activated DC induce differentiation of human T cells into potent Th1 effectors. Cocultivation of autologous T cells and CD137L-activated DC in an antigen-specific reaction leads to T cell proliferation and the release of IL-12p70 and IFN-gamma. These findings deliver new insights into the multiple effects of reverse signaling of CD137L in human DC during the initiation of an adaptive immune response, including the key features of DC maturation, migration and, ultimately, antigen specific T cell differentiation. PMID- 18395855 TI - Regulatory T cells in the past and for the future. PMID- 18395856 TI - Regulatory T cells and immune computation. AB - The role of Treg in immune regulation is the topic of this Viewpoint series in the European Journal of Immunology (EJI); the question to be discussed in this section is the effector function of Treg in immune regulation. In this manuscript, we take on the following three postulates outlined by Rolf Zinkernagel on the role of Treg in the control of immunity. First, the immune response is regulated primarily by the antigen and not by Treg. Second, immune non-responsiveness results from the deletion of specific receptor-bearing T cells. Third, there is no definitive proof of the existence of specialized Treg that know what is needed for an equilibrated immune response. Herein, we discuss data demonstrating the existence of specialized Treg and therefore arguing against the validity of the first two postulates. However, based on the reactive nature of the immune system, we agree with Rolf's third postulate in that Treg cannot know ahead of time an ideal set-point for immune homeostasis. PMID- 18395857 TI - How many mechanisms do regulatory T cells need? AB - A plethora of new regulatory T cell (Treg) mechanisms have recently emerged. This raises two important questions. First, how many molecules or mechanisms are required for Treg to work? Second, how should we evaluate the contribution of any given Treg molecule/mechanism and how is this likely to relate (or not) to the phenotype seen in Scurfy/Foxp3-deficient mice? In this discussion piece, I will briefly outline our current understanding of the Treg arsenal and address these important questions. PMID- 18395858 TI - Critical role of IL-2 and TGF-beta in generation, function and stabilization of Foxp3+CD4+ Treg. AB - CD4+Foxp3+ Treg consist of two indistinguishable subsets induced in either the thymus or the periphery. In addition to their suppressive activities, IL-6 can convert natural Treg to pro-inflammatory IL-17-producing cells, but those induced with IL-2 and TGF-beta remain Treg. Unlike mouse CD4+CD25(-) cells, which rapidly become polyclonal Foxp3+CD25+ Treg when activated appropriately with IL-2 and TGF beta, human T cells require multiple stimulations to become similar suppressor cells. PMID- 18395859 TI - The critical contribution of TGF-beta to the induction of Foxp3 expression and regulatory T cell function. AB - Stimulation of mouse CD4+ T cells in the presence of TGF-beta results in the expression of Foxp3 and induction of Treg function. Stimulation of human CD4+ T cells under similar conditions results in the expression of Foxp3, but the cells lack regulatory cell function. TGF-beta expressed on the surface of Treg also induces Foxp3 expression and Treg function in responder cells. Both of these mechanisms may play a role in vivo in the induction of antigen-specific extra thymic Treg. PMID- 18395860 TI - Role of Foxp3-positive regulatory T cells during infection. AB - Surviving an infection requires the generation of an immune response that controls the invading pathogen while limiting collateral damage to self tissues that may result from an exuberant immune response. Various populations of regulatory cells, including Foxp3+ Treg, have been shown to play a central role in the establishment of these controlled immune responses. In this review, I discuss current hypotheses and points of polemic associated with the origin, mode of action and antigen specificity of Foxp3+ Treg during infection. PMID- 18395861 TI - Human regulatory T cells and autoimmunity. AB - CD4+CD25+ regulatory T cells (Treg) appear to be critical in regulating immune responses to self-antigens. Treg deficiency is associated with several human autoimmune diseases. Although substantial progress has been made in the study of murine and human Treg, their fundamental mechanism of action remains unknown. In this review, we discuss the phenotype of human natural Treg, their functional mechanism, and their role in autoimmune disease. PMID- 18395862 TI - Is FOXP3 a bona fide marker for human regulatory T cells? AB - FOXP3 is a specific marker for naturally occurring regulatory T cells (nTreg). FOXP3 expression is correlated with development and function of nTreg. Recent evidence suggests that, upon activation, human effector T (Teff) cells and type 1 regulatory T (Tr1) cells can express FOXP3, albeit transiently. While the role of transient FOXP3 expression in Teff cells is poorly understood, it is becoming clear that it does not correlate with suppressor function. Furthermore, FOXP3 independent mechanisms, mediated by IL-10, contribute to the induction and suppressor functions of Tr1 cells. PMID- 18395863 TI - Rethinking the molecular definition of regulatory T cells. AB - It has been postulated that expression of the transcription factor Foxp3 "specifically" identifies the regulatory T cell (Treg) lineage and it acts as its "master regulator" or "lineage specification factor". However, emerging evidence challenges these views and suggests that the Treg lineage may be determined by a higher-order regulatory process that ensures stable and high levels Foxp3 expression. PMID- 18395864 TI - Clinical application of regulatory T cells for treatment of type 1 diabetes and transplantation. AB - Immune regulation is a complex process that depends on the maintenance of self tolerance while retaining robust immune responses against microbes. The emergence of Treg as a central mechanism for immune regulation has generated a new paradigm where Treg-resistant memory T cells and/or "defective" Treg lead to a breakdown in tolerance resulting in immune pathology. In this perspective, we highlight the opportunities and challenges in the field of Treg therapy. PMID- 18395865 TI - Inhibin A measurement using an automated assay platform. AB - BACKGROUND: Second-trimester measurement of maternal serum inhibin A is widely used for Down syndrome screening. To date, only a manual enzyme-linked immunosorbent assay (ELISA) produced by Diagnostic Systems Laboratories, Inc (DSL) has been available. The objective of this study was to compare the DSL assay with a new automated assay produced by Beckman Coulter, Inc (Access). METHODS: Residual serum samples from 570 women, who were receiving routine screening for Down syndrome, were retrieved from storage. The Access assay sensitivity, linearity and reproducibility were determined and a method comparison was performed. Inhibin A levels were measured using both assays. Twenty samples from women with confirmed Down syndrome pregnancy were also tested. RESULTS: The Access assay had coefficients of variation of less than 10% across the range of values tested, and a sensitivity below 1 pg/mL. The DSL and Access inhibin A assay values were highly correlated (r = 0.961, r(2) = 0.923), with no apparent outliers. Inhibin A values from the Access assay were a constant 23% lower (95% CI 1-41%) than corresponding values from the DSL assay. Median values from 15 to 20 completed weeks' gestation were computed and found to be consistent with expectations. The weight-adjusted multiples of the median (MoM) levels in the unaffected pregnancies fit a log Gaussian distribution well between at least the 5th and 95th percentiles with corresponding log standard deviations of 0.1960 and 0.1919 for DSL and Access, respectively. CONCLUSIONS: With median inhibin A levels appropriately calculated for the Access assay, Down syndrome screening performance is expected to be comparable to that obtained with the manual DSL assay. PMID- 18395866 TI - Clinico-pathologic features and outcome of Japanese patients with peripheral T cell lymphomas. AB - We studied the clinico-pathologic features and treatment outcome of patients with peripheral T-cell lymphoma (PTCL). This study included 215 patients with T/natural killer (NK)-cell lymphoma, including 59 with PTCL-unspecified (PTCL-U), 42 with angioimmunoblastic T-cell lymphoma (AILT) and 20 with anaplastic large cell lymphoma (ALCL). Most of the analyses were performed on patients with AILD, ALCL and PTCL-U. The patients with AILT and PTCL-U tended to be older than those with ALCL. Stage III/IV disease was seen in 90.5% of the AILT cases, 55% of the ALCL cases and 67.8% of the PTCL-U cases. In addition, 61.9% of the AILT cases had an international prognostic index (IPI) of H-I or H risk. The 5-year progression-free survival (PFS) and overall survival (OS) rates were 72.2 and 76.1% among the ALCL cases, 40.7 and 42.2% among the PTCL-U cases and 31.2 and 49.3% among the AILT cases, respectively. Among the patients with PTCL-U, the 5 year PFS and OS rates in group low (L), low-intermediate (L-I), high-intermediate (H-I) or high (H) risk group of IPI were: 47.6 and 56.1%, 55.6 and 53.8%, 42.4 and 40.1% and 9.1 and 9.1%, respectively. The 5-year PFS and OS rates in group 1, 2, 3 or 4 by prognostic index of PTCL-U (PIT) were: 88.9 and 85.7%, 57.1 and 54.9%, 33.5 and 28.8% or 13.3 and 13.3%, respectively. The 5-year PFS and OS rates among patients who received CHOP therapy, CyclOBEAP [cyclophosphamide (CPA), vincristine (VCR), bleomycine, etoposide, doxorubicin (DXR), prednisone (PDN)] therapy or autologous stem cell transplantation were: 22 and 25.7%, 59 and 61.7% or 33.3 and 60%, respectively. Multivariate analysis revealed that the PIT score was associated with OS and PFS. These results indicate that the presence of bone marrow (BM) involvement is an independent prognostic factor which may predict both OS and PFS. PTCL-U is a heterogeneous disease with regard to histological type and pathological state. Because PTCL-U is generally not responsive to CHOP therapy, new treatment strategies need to be developed. PMID- 18395867 TI - The diabetic foot in the developing world. AB - This short article provides a brief overview of the epidemiology of diabetes and the diabetic foot in developing countries, reviews evidence that diabetic foot care is potentially one of the most cost-effective interventions for people with diabetes in developing countries, and outlines some of the challenges to improving access to foot care and diabetes care in general. PMID- 18395868 TI - Analysis of multiple loci can increase reliability of detection of fetal Y chromosome DNA in maternal plasma. AB - OBJECTIVE: Aiming to develop more reliable methods for determination of fetal gender from maternal plasma we compared three different systems of polymerase chain reaction (PCR) detection of Y-chromosome DNA. METHODS: Cell-free DNA was isolated from 96 samples of maternal plasma and (1) amplified using AmpFLSTR Identifiler (15 autosomal STR loci and amelogenin) or AmpFLSTR-Yfiler (16 Y chromosome STR loci) kits and subsequently analyzed on ABI-PRISM 310 Genetic Analyzer, or (2) analyzed using Quantifiler-Y DNA-Quantification kit. Gender of fetuses was confirmed by cytogenetic analysis or phenotypically at birth. RESULTS AND CONCLUSIONS: AmpFLSTR-Identifiler and Quantifiler-Y Human-Quantification kits were rather reliable in determining fetal gender (92.5 and 98.1%, respectively), but false negatives were still present in both systems. AmpFLSTR-Yfiler was found to be fully reliable as it amplified Y-chromosome in all cases of male fetuses, and was thus 100% correct in determining fetal gender. In addition, it enabled comparison of polymorphic Y-chromosome loci between father and a child, thus further supporting specificity of obtained results. PMID- 18395869 TI - Anxiety levels in women undergoing prenatal maternal serum screening for Down syndrome: the effect of a fast reporting system by mobile phone short-message service. AB - OBJECTIVE: To study the effect of fast reporting by mobile phone short-message service (SMS) on anxiety levels in women undergoing prenatal biochemical screening for Down syndrome. METHOD: From January 2005 to December 2006, 2782 women undergoing prenatal biochemical serum screening were randomized into fast reporting by SMS (group A) or without mobile phone reporting (group B). Anxiety levels were measured with the Spielberger State-Trait Anxiety Inventory (STAI) before prenatal screen testing, before the appointed clinic (when the SMS report had already been given to group A), and 3 days after the appointed clinic (when the full screening report had been given to groups A and B). RESULTS: For screen negative women, anxiety scores did not differ between groups before prenatal screen testing and 3 days after the appointed clinic. The state-anxiety scores measured on the second occasion had declined significantly in group A. The state anxiety scores in both groups increased over the 3-week period after being informed of positive screen results. The trait- and state-anxiety scores at all points did not differ between the two groups of screen-positive women. CONCLUSIONS: The provision of a routine reporting system plus additional SMS report revealed some overall benefits in reducing anxiety among women with screen negative result. PMID- 18395870 TI - First trimester prenatal diagnosis of 13q-syndrome presenting with increased nuchal translucency, Dandy-Walker malformation and small parietal encephalocoele. PMID- 18395871 TI - New developments in diagnosing and treating diabetic foot infections. AB - Foot infections are common in persons with diabetes and are often the proximate cause of lower extremity amputation. There have been many publications in the past few years dealing with the appropriate ways to diagnose and treat diabetic foot infections. This review presents information gathered from a comprehensive, ongoing surveillance of the literature (published and abstracts) over the past 4 years. Prospective studies have now defined the epidemiology of diabetic foot infections, as well as methods to score and classify the wounds. Several recently published guidelines can assist clinicians in managing these infections. The etiologic agents of infection have been well-defined, and the prevalence of multi drug-resistance pathogens is growing. Molecular methods offer great promise for quicker and more sensitive diagnosis of infection. New antimicrobial agents, both systemic and topical, as well as novel local treatments, have been shown to be effective in various studies. Improved methods of deploying older agents have added to the variety of treatment approaches now available. Several adjunctive treatments may benefit some patients but their role is as yet unclear. While there is much yet to learn about the most cost-effective ways to diagnose and treat diabetic foot infections the main effort is now to disseminate the available information and facilitate employing the evidence-based guideline recommendations. PMID- 18395872 TI - Refined fluorescent STR quantification of cell-free fetal DNA during pregnancy in physiological and Down syndrome fetuses. AB - BACKGROUND: Cell-free fetal (cff) DNA analysis by short tandem repeats (STR) has the advantage of better recognizing the different genotypes. However, quantitative examination by quantitative fluorescent (QF) polymerase chain reaction (PCR) by STRs is limited to only a rough approximation. This project focuses on a more precise calculation of the relative cff DNA amount tested in the STRs' loci. METHODS: The cff DNA was analyzed in 363 samples from 258 pregnant women with physiological fetuses in different stages of pregnancy (from 4-37 gestational weeks) separately in three STRs [D21S1435, D21S1446 and PentaD (pD)] and also by gonosomal sequences amelogenin gene, X/Y-linked/testis specific protein, Y-linked (AMELX/Y/TSPY). Seventeen samples of cff DNA from fetuses with Down syndrome (DS) were compared. We optimized the refined quantitative fluorescent (RQF) PCR for STRs in a particular locus. RESULTS AND CONCLUSIONS: The cff DNA detection rate was 74% in at least one of the STRs. The efficiency decreased from shorter to longer PCR fragments. All three STR and gonosomal loci proved an increase in cff DNA during pregnancy. The stutter variability rate is greatest in short STR fragments and decreases as the STR fragments increase in length. Results showed that DS samples had a significantly higher amount of cff DNA. PMID- 18395873 TI - Maternal serum ADAM12s in the late first trimester of pregnancies with Trisomy 21. AB - BACKGROUND: ADAM12s is a placenta-derived glycoprotein that is involved in growth and differentiation, and has been shown to be a potential first-trimester and second-trimester marker of Trisomy 21 and other aneuploides. Maternal ADAM12s concentrations show a considerable temporal variation with gestational age and here we study the levels at 11-13 weeks of gestation to establish the effectiveness or otherwise at a time when other established markers are used. MATERIALS AND METHODS: Samples collected as part of routine first-trimester screening were retrieved from storage. In total, 46 samples from pregnancies with Trisomy 21 were identified and collected between 11 and 13 weeks of gestation-of these 83% had been identified by combined first-trimester screening. A series of 414 gestational age-matched samples collected during the same period formed the control group. ADAM12s was measured by a new DELFIA assay incorporating two monoclonals (6E6 and 8F8). Results were expressed as weight-corrected multiples of the median (MoM). RESULTS: The median MoM ADAM12s rose from 0.914 at 11 weeks to 1.032 at 13 weeks. CONCLUSIONS: Combining the data from this study and other published studies suggests that ADAM12s is unlikely to be of much additional value when screening for Trisomy 21 in the period 11-13 weeks. More studies are required looking at the potential of ADAM12s prior to 10 weeks. PMID- 18395874 TI - Pregnancy outcome following prenatal diagnosis of sex chromosome abnormalities in Mainland China. PMID- 18395875 TI - Inherited unbalanced structural chromosome abnormalities at prenatal chromosome analysis are rarely ascertained through recurrent miscarriage. AB - OBJECTIVE: To determine the mode of ascertainment of inherited unbalanced structural chromosome abnormalities detected at prenatal chromosome analysis. METHODS: From the databases of three centres for clinical genetics in the Netherlands, all cases of inherited unbalanced structural chromosome abnormalities detected at prenatal chromosome analysis in the period 1992-2000 were selected. The mode of ascertainment was identified by examining the reason for prenatal chromosome analysis and the reason for parental chromosome analysis of the first structural chromosome abnormality detected within the family. RESULTS: Totally 56 cases of inherited unbalanced structural chromosome abnormalities were detected at prenatal chromosome analysis. Only one case was ascertained through two previous miscarriages (2%). The main modes of ascertainment were a previous child with an unbalanced karyotype (48%), congenital abnormalities at ultrasound examination (20%), and advanced maternal age (9%). The remaining cases had a different mode of ascertainment. CONCLUSION: Inherited unbalanced structural chromosome abnormalities detected at prenatal chromosome analysis are rarely ascertained through two or more miscarriages. PMID- 18395876 TI - Prenatal testing for a novel EBP missense mutation causing X-linked dominant chondrodysplasia punctata. AB - OBJECTIVE: To determine the pathogenicity of a novel conserved missense mutation, p.Ser98Phe, in the emopamil binding protein (EBP) gene in order to perform a prenatal diagnostic test for X-linked dominant chondrodysplasia punctata (CDPX2) in a male foetus at 50% risk. METHODS: Family members were tested for p.Ser98Phe using PCR and sequence analysis of leucocyte or buccal cell DNA. Haplotype analysis was employed to identify the grandparental chromosome on which p.Ser98Phe had arisen. In silico protein analyses were used to predict whether p.Ser98Phe significantly altered the EBP protein structure. RESULTS: The mutation was detected in the proband and her affected mother but not in the maternal grandmother, who was thought to be mildly affected. However, haplotype analysis showed that p.Ser98Phe had arisen de novo on the grandpaternal X chromosome. Protein secondary structure predictions suggested that p.Ser98Phe alters the properties of the helix within which it is located. CONCLUSION: We concluded that p.Ser98Phe is likely to be pathogenic and proceeded with prenatal testing. The male foetus had not inherited p.Ser98Phe and his unaffected status was confirmed at birth. This family demonstrates some of the difficulties in interpreting the significance of novel missense mutations, particularly when results are needed urgently for prenatal diagnosis. PMID- 18395877 TI - Familial Beckwith-Wiedemann syndrome due to CDKN1C mutation manifesting with recurring omphalocele. PMID- 18395878 TI - Fetal genital anomalies: an aid to diagnosis. AB - OBJECTIVE: To report our experience with the prenatal diagnosis of fetal genital anomalies and suggest a protocol for management. METHODS: A retrospective review of all the cases with fetal genital anomalies or phenotype and genotype discrepancy identified by prenatal ultrasound. RESULTS: Twenty cases with abnormal fetal genitalia and four with a phenotype and genotype discrepancy were diagnosed prenatally. Genital anomalies were rarely found in isolation, most were found in combination with renal or multiple structural anomalies. The etiology of abnormal genitalia was broad and included metabolic, chromosomal and genetic syndromes. CONCLUSION: Prenatal detection of genital anomalies should stimulate a detailed ultrasound examination and determination of genotypic sex. Measurement of 17-OHP and Delta(4)-androstenedione or metabolites of the cholesterol pathway in the amniotic fluid and/or maternal urine may be helpful in making a definitive diagnosis. Identification of genital anomalies in fetuses with renal or multiple abnormalities can aid prenatal diagnosis, thereby facilitating accurate counseling of parents who are then in a better position to make informed choices. PMID- 18395879 TI - Prenatal diagnosis of concomitant Wolf-Hirschhorn syndrome and split hand-foot malformation associated with partial monosomy 4p (4p16.1-->pter) and partial trisomy 10q (10q25.1-->qter). PMID- 18395880 TI - A cross-sectional survey of herbal remedy taking by United Arab Emirate (UAE) citizens in Abu Dhabi. AB - PURPOSE: To determine patterns of United Arab Emirates (UAE) citizen herbal medicine taking. METHODS: Questionnaire survey of UAE citizens attending a primary healthcare clinic in Abu Dhabi. RESULTS: Three hundred and thirty surveys were analysed. Seventy-six per cent (250/330) had previously used herbal products, and of these 38% (94/248) were currently taking at least one herb. In total, 65 different herbs were being taken to treat 48 conditions. Respondents overwhelmingly felt herbal medicines were safe, yet 27 people reported experiencing an adverse drug reaction. Causality was not established because of multiple herbal use and a propensity for concurrent herbal and Western medicine taking. Respondents relied heavily on family and friends for advice and bought herbs from a 'condimental shop' (local herbal seller/healer). Despite high herbal use, respondents still preferred Western medicine to treat chronic diseases. CONCLUSIONS: This study has shown that UAE nationals have faith and confidence in herbal medicines and are consequently high users of them despite the country's progressive adoption of Western approaches to health care. PMID- 18395881 TI - Structure elucidation of five new iridoid glucosides from the leaves of Avicennia marina. AB - A chemical examination of the mangrove plant Avicennia marina (Forrsk.) Vierh resulted in the isolation and characterization of five new iridoids, marinoids A E (1-5), along with 2'-cinnamoyl-mussaenosidic acid (6), 2'-O-(4'' methoxycinnamoyl) mussaenosidic acid (7), 2'-O-(4''-hydroxycinnamoyl) mussaenosidic acid (8), and 3(R)-hydroxy-5-phenyl-4(E)-pentenoic acid (9). The structures of 1-5 were elucidated on the basis of 1D and 2D NMR (COSY, HMQC, HMBC, and NOESY) in association with IR and MS data analysis. PMID- 18395882 TI - The 'hybrid cell': a new compensated infinity cell for larger radius ion excitation in Fourier transform ion cyclotron resonance mass spectrometry. AB - A new 'hybrid' ion cyclotron resonance (ICR) trap is proposed and analyzed by computer simulations. The trap is basically a hybrid of a segmented end cap (Infinity) and capacitively coupled cylindrical cell, with additional electrodes placed at the center of each end cap. The new trap produces an on-axis electric field z-profile similar to that of the Infinity cell or capacitively coupled open cylindrical cell during ion excitation. Simion simulations demonstrate that, during detection, appropriate changes of the potentials applied to the two new sets of electrodes produce a radial electric field z-profile that more closely approaches that for an ideal axial three-dimensional quadrupolar potential at high post-excitation ICR orbital radius, for improved signal-to-noise ratio and resolving power, and minimal m/z-discrimination. PMID- 18395883 TI - Adherence to the immunomodulatory drugs for multiple sclerosis: contrasting factors affect stopping drug and missing doses. AB - BACKGROUND: Long-term immunomodulatory drug (IMD) treatment is now common in multiple sclerosis (MS). However, predictors of adherence are not well understood; past studies lacked lifestyle factors such as alcohol use and predictors of missed doses have not been evaluated. We examined both levels of non-adherence-stopping IMD and missing doses. METHODS: This longitudinal prospective study followed a population-based cohort (n = 199) of definite MS patients in Southern Tasmania (January 2002 to April 2005, source population 226 559) every 6 months. Baseline factors (demographic, clinical, psychological and cognitive) affecting adherence were examined by logistic regression and a longitudinal analysis (generalized estimating equation (GEE)). RESULTS: Of the 97 patients taking an IMD (mean follow-up = 2.4 years), 73% (71/97) missed doses, with 1 in 10 missing > 10 doses in any 6-month period. Missed doses were positively associated with alcohol amount consumed per session (p = 0.008). A history of missed doses predicted future missed doses (p < 0.0005). Over one quarter (27/97) stopped their current IMD, which was associated with lower education levels (p = 0.032) and previous relapses (p = 0.05). No cognitive or psychological test predicted adherence. CONCLUSIONS: There were few strong predictors of missed doses, although people with MS consuming more alcoholic drinks per session are at a higher risk of missing doses. Divergent factors influenced the two levels of non-adherence indicating the need for a multifaceted approach to improving IMD adherence. In addition, missed doses should be assessed and incorporated into clinical trial design and clinical practice as poor adherers could impact on clinical outcomes. PMID- 18395884 TI - Monosize polycationic nanoparticles as non-viral vectors for gene transfer to HeLa cells. AB - Monosized cationic nanoparticles were produced by emulsifier-free emulsion polymerization of styrene, poly(ethylene glycol) methacrylate and N-[3-(dimethyl amino) propyl] methacrylamide, conducted in the presence of a cationic initiator, 2,2'-azobis (2-methylpropionamidine) dihydrochloride, using different amounts of ingredients and at different conditions. The structure of the terpolymers was confirmed by (1)H-NMR and FTIR spectroscopy. The nanoparticles, with an average size of 71.3 nm [polydispersity index (PDI), 1.110] and a Zeta potential of 65.6 mV obtained by a zeta sizer, were used in the transfection studies. HeLa cells were transfected in in vitro cell cultures with these non-viral nanoparticle vectors with a green fluorescent protein (GFP)-expressing plasmid DNA. The transfer of the cationic nanoparticles into the cells and GFP expressions with the conjugates of the nanoparticles and the GFP-expressing plasmid were followed by both light and fluorescent microscopy. The GFP expression efficiency was unexpectedly high (up to 90%). PMID- 18395885 TI - The Frontal Systems Behavior Scale in the evaluation of dementia. PMID- 18395886 TI - Depression in elderly life sentence prisoners. AB - BACKGROUND: The life sentence population is growing older and increasing in number. Despite the potential negative physical and social environment in prisons little is known about the prevalence or aetiology of depression in elderly 'lifers'. AIMS: To determine the prevalence and associated risk factors of depression in elderly life sentence prisoners. METHOD: One hundred and twenty-one elderly life/indeterminate sentence prisoners from two category B prisons in the United Kingdom were interviewed using the Geriatric Depression Scale and the relationship with prison and non prison specific variables analysed. RESULTS: Over half of the prisoners scored above the threshold for mild depression. The length of sentence served and other prison related variables were not associated with the depression score. However, the imported chronic physical ill health was strongly related to depression score. CONCLUSIONS: Depression in long term prisoners is common and is related to the burden of imported chronic ill health as opposed to specific effects of imprisonment. PMID- 18395887 TI - The frequency of cognitive dysfunction in elderly Sri Lankans with type 2 diabetes mellitus. PMID- 18395888 TI - Total serum cholesterol levels and Alzheimer's dementia in patients with Down syndrome. AB - BACKGROUND: The risk for dementia in Alzheimer's disease (DAD) in adults with Down syndrome (DS) is higher than in the general adult population. Hypercholesterolaemia has been reported as a risk factor for DAD in the general population. This study investigated the role of serum cholesterol levels in the onset of DAD in the DS population. METHODS: This study investigated total serum cholesterol levels in 179 DS persons (with and without DAD). The possible association between Apolipoprotein E and amyloid beta1-40 and beta1-42 levels was also investigated. RESULTS: No statistically significant association was found between total serum cholesterol levels and dementia in AD or with amyloid beta levels. However for DS adults with an apoE epsilon4 allele significantly higher serum cholesterol levels were found. CONCLUSION: Hypercholesterolaemia is not a risk factor for DAD in persons with DS. However, DS persons with an apoE epsilon4 allele are susceptible to high serum cholesterol. Such individuals should be screened on a regular basis. PMID- 18395889 TI - Longitudinal assessment of psychotherapeutic day hospital treatment for elderly patients with depression. AB - OBJECTIVE: Although previous studies suggested that psychiatric day hospital care is a valuable alternative to inpatient treatment, its effectiveness for elderly patients is disputed. Small number of cases, poor definition of the psychotherapeutic setting, and absence of systematic assessment at different time points may explain the observed discrepancies. We performed an assessment of a psychiatric day hospital treatment combining individual and group psychotherapy in a series of 122 elderly depressed outpatients. METHODS: The Geriatric Depression Scale, Short Form Survey, as well as a Therapeutic Community Assessment Scale and Group Evaluation Scale were repeated at admission, 3, 6, 12 months and discharge. The day hospital program was based on psychotherapeutic treatment combining individual and group settings. All patients presented with major depression or a depressive episode of bipolar disease. Variables included severity of depressive symptoms, quality of life, adhesion to therapeutic community treatment and progress in groups of psychotherapy, art-therapy, and psychomotricity. RESULTS: There was a significant reduction of depressive symptoms, and improvement in mental quality of life across all time points studied. Adhesion to therapeutic community increased from admission to discharge. This was also the case for the progress in group therapy for all three groups used, yet the evolution of this parameter at intermediate time points was highly variable. Neither demographic characteristics, nor pharmacological treatment or presence of stressful life events predicted the clinical improvement. CONCLUSIONS: Psychotherapeutic care program in day hospitals may improve clinical status and quality of life in elderly depressed patients. PMID- 18395890 TI - Painful diabetic neuropathy: treatment and future aspects. AB - Around one of three diabetic patients is affected by distal symmetric polyneuropathy (DSP) which represents a major health problem, as it may present with partly excruciating neuropathic pain and is responsible for substantial morbidity and increased mortality. Treatment is based on four cornerstones: (1) multifactorial intervention aimed at (near)-normoglycaemia and reduction in cardiovascular risk factors, (2) treatment based on pathogenetic mechanisms, (3) symptomatic treatment, and (4) avoidance of risk factors and complications. Among the pathogenetic treatments only alpha-lipoic acid and epalrestat are available for treatment in several countries. Neuropathic pain, which is present in 8-26% of diabetic patients, exerts a substantial impact on the quality of life, particularly by causing considerable interference in sleep and enjoyment of life. Non-pharmacologic options such as nerve or muscle stimulation should always be given consideration. Among the centrally acting analgesic drugs for many years mainly the tricyclic antidepressants (TCA), carbamazepine, gabapentin, and opioids have been used to treat neuropathic pain. More recently, significant pain relief has been reported in clinical trials of painful diabetic neuropathy using agents such as the dual selective serotonin noradrenaline reuptake inhibitor (SNRI), duloxetine and the anticonvulsant pregabalin, a specific modulator of the alpha(2)delta subunit of the voltage-dependent calcium channels. A promising new anticonvulsant is lacosamide. In future, drug combinations might also include those aimed at symptomatic pain relief and quality of life on one hand and improvement or slowing the progression of the underlying neuropathic process on the other hand. PMID- 18395891 TI - Reverse micellar microextraction for rapid analysis of thiol-containing peptides and amino acids by atmospheric-pressure matrix-assisted laser desorption/ionization ion trap and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Simple, rapid and inexpensive one-step reverse micellar microextraction (RMME) procedures were combined with matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for the determination of thiol-containing peptides and amino acids. In this investigation, a thiol-containing peptide (HW6) was chosen as model compound to understand the mechanism of RMME. The electrostatic interactions between the thiol-containing peptide and reverse micelles were proposed to be reason for the transfer of analytes from the aqueous phase to the organic phase. Reverse micelles were formed by the cationic surfactant, methyltrioctylammonium chloride (MTOAC). The best extraction efficiency of HW6 was obtained under the following conditions: pH 11.0, ionic strength 5.0 mM of KCl and micelle concentration 7.0 mM of MTOAC. The limits of detection (LODs) obtained for HW6 in water, urine and plasma samples were 0.15, 0.19 and 0.28 microM, respectively, with relative standard deviation (RSD) values in the range +/-8.8-10.5%. The sensitivity obtained in water by the present method was 45-fold higher than that of the conventional use of atmospheric-pressure (AP)-MALDI MS. Furthermore, the applicability of the proposed approach was extended for the determination of thiol-containing amino acids in sample solutions by using MALDI time-of-flight (TOF) MS. PMID- 18395892 TI - Reply to the letter to the editor "Long-term cannabinoid receptor (CB1) blockade in obesity: implications for the development of colorectal cancer". PMID- 18395893 TI - Wireless capsule endoscopy. AB - Wireless capsule endoscopy is a new technique that allows complete exploration of the small bowel without external wires. Its role has been analyzed in many small bowel diseases such as obscure gastrointestinal bleeding, Crohn's disease and gastrointestinal polyposis syndromes with promising results. Studies on other pathologies (i.e. small bowel tumour, celiac disease) are under evaluation to define the role of this technique. PMID- 18395894 TI - Current view of the immunopathogenesis in inflammatory bowel disease and its implications for therapy. AB - Al though the aetiology of inflammatory bowel disease (IBD) remains unknown, the pathogenesis is gradually being unravelled, seeming to be the result of a combination of environmental, genetic, and immunological factors in which an uncontrolled immune response within the intestinal lumen leads to inflammation in genetically predisposed individuals. Multifactorial evidence suggests that a defect of innate immune response to microbial agents is involved in IBD. This editorial outlines the immunopathogenesis of IBD and their current and future therapy. We present IBD as a result of dysregulated mucosal response in the intestinal wall facilitated by defects in epithelial barrier function and the mucosal immune system with excessive production of cytokines growth factors, adhesion molecules, and reactive oxygen metabolites, resulting in tissue injury. Established and evolving therapies are discussed in the second part of this editorial and at the end of this section we review new therapies to modulate the immune system in patients with IBD. PMID- 18395895 TI - Role of endoscopy in the management of acute diverticular bleeding. AB - Colonic diverticulosis is one of the most common causes of lower gastrointestinal bleeding. Endoscopy is not only a useful diagnostic tool for localizing the bleeding site, but also a therapeutic modality for its management. To date, haemostatic methods have included adrenaline injection, mechanical clipping, thermal and electrical coagulation or combinations of them. The results of all published data are herein reviewed. PMID- 18395896 TI - Endoscopic mucosal resection in the upper gastrointestinal tract. AB - Endoscopic mucosal resection (EMR) is a technique used to locally excise lesions confined to the mucosa. Its main role is the treatment of advanced dysplasia and early gastrointestinal cancers. EMR was originally described as a therapy for early gastric cancer. Recently its use has expanded as a therapeutic option for ampullary masses, colorectal cancer, and large colorectal polyps. In the Western world, the predominant indication for EMR in the upper gastrointestinal tract is the staging and treatment of advance dysplasia and early neoplasia in Barrett's esophagus. This review will describe the basis, indications, techniques, and complications of EMR, and its role in the management of Barrett's esophagus. PMID- 18395897 TI - Relationship between cell adhesion molecules expression and the biological behavior of gastric carcinoma. AB - AIM: To evaluate the relationship between the expression of cell adhesion molecules (CAMs) and the biological behavior of gastric carcinoma. METHODS: Expression of syndecan-1, E-cadherin and integrin beta3 were evaluated by immunohistochemical study in a total of 118 gastric carcinomas and 20 non-tumor gastric mucosas. RESULTS: The expressions of syndecan-1 and E-cadherin were significantly lower in gastric carcinoma compared to non-tumor gastric mucosa, and the low expression rates were positively correlated to the tumor invasion depth, vessel invasion, lymph node metastasis and distant metastasis (P < 0.01 in all cases). However, the expression of integrin beta3 was significantly higher in gastric carcinoma compared to non-tumor gastric mucosa, and the high expression rates were positively correlated to the tumor invasion depth, vessel invasion, lymph node metastasis and distant metastasis (P < 0.01 in all cases). In addition, the three protein expressions were correlated to the tumor growth pattern (P < 0.01, P < 0.01, and P < 0.05 respectively), but not correlated to tumor differentiation (P > 0.05, P > 0.05 and P > 0.05 respectively). Positive correlation was observed between the expressions of syndecan-1 and E-cadherin, but they which were negatively correlated to the expression of integrin beta3 (P < 0.01 in all cases). Univariate analysis demonstrated that the mean survival time and 5-year survival rate were lower in the cases with low expressions of syndecan-1 and E-cadherin and high expression of integrin beta3 (P < 0.01, in all cases). COX multivariate analysis showed that the expression level of syndecan-1 could be an independent prognostic index of gastric carcinoma (P < 0.01), whereas E-cadherin and integrin beta3 could not be independent indexes (P > 0.05, P > 0.05 respectively). CONCLUSION: The low expression of syndecan-1 and E-cadherin and the high expression of integrin beta3 are significantly correlated with the invasion and metastasis of gastric carcinoma, and they are highly correlated with each other. Therefore they may serve as important prognostic markers of gastric carcinoma. PMID- 18395898 TI - Long-term outcome of percutaneous ethanol injection therapy for minimum-sized hepatocellular carcinoma. AB - AIM: To evaluate long-term follow-up of minimum-sized hepatocellular carcinoma (HCC) treated with percutaneous ethanol injection (PEI). METHODS: PEI was applied to 42 lesions in 31 patients (23 male and eight female) with HCC < 15 mm in diameter, over the past 15 years. RESULTS: Overall survival rate was 74.1% at 3 years, 49.9% at 5 years, 27.2% at 7 years and 14.5% at 10 years. These results are superior to, or at least the same as those for hepatic resection and radiofrequency ablation. Survival was affected only by liver function, but not by sex, age, etiology of Hepatitis B virus or Hepatitis C virus, alpha-fetoprotein levels, arterial and portal blood flow, histological characteristics, and tumor multiplicity or size. Patients in Child-Pugh class A and B had 5-, 7- and 10 years survival rates of 76.0%, 42.2% and 15.8%, and 17.1%, 8.6% and 0%, respectively (P = 0.025). CONCLUSION: Treatment with PEI is best indicated for patients with HCC < 15 mm in Child-Pugh class A. PMID- 18395899 TI - Anti-cancer and anti-angiogenic effects of curcumin and tetrahydrocurcumin on implanted hepatocellular carcinoma in nude mice. AB - AIM: To determine the effect of tetrahydrocurcumin (THC) on tumor angiogenesis compared with curcumin (CUR) by using both in vitro and in vivo models of human hepatocellular carcinoma cell line (HepG2). METHODS: The 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was used for testing the anti proliferating activities of CUR and THC. In male BALB/c nude mice, 2 multiply 10(6) human HepG2 cells were inoculated onto a dorsal skin-fold chamber. One day after HepG2 inoculation, the experimental groups were fed oral daily with CUR or THC (300 mg/kg or 3000 mg/kg). On d 7, 14 and 21, the tumor microvasculature was observed using fluorescence videomicroscopy and capillary vascularity (CV) was measured. RESULTS: Pathological angiogenic features including microvascular dilatation, tortuosity, and hyper-permeability were observed. CUR and THC could attenuate these pathologic features. In HepG2-groups, the CV were significantly increased on d 7 (52.43%), 14 (69.17%), and 21 (74.08%), as compared to controls (33.04%, P < 0.001). Treatment with CUR and THC resulted in significant decrease in the CV (P < 0.005 and P < 0.001, respectively). In particular, the anti angiogenic effects of CUR and THC were dose-dependent manner. However, the beneficial effect of THC treatment than CUR was observed, in particular, from the 21 d CV (44.96% and 52.86%, P < 0.05). CONCLUSION: THC expressed its anti angiogenesis without any cytotoxic activities to HepG2 cells even at the highest doses. It is suggested that anti-angiogenic properties of CUR and THC represent a common potential mechanism for their anti-cancer actions. PMID- 18395900 TI - Serial changes in expression of functionally clustered genes in progression of liver fibrosis in hepatitis C patients. AB - AIM: To investigate the relationship of changes in expression of marker genes in functional categories or molecular networks comprising one functional category or multiple categories in progression of hepatic fibrosis in hepatitis C (HCV) patients. METHODS: Marker genes were initially identified using DNA microarray data from a rat liver fibrosis model. The expression level of each fibrosis associated marker gene was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) in clinical biopsy specimens from HCV-positive patients (n = 61). Analysis of changes in expression patterns and interactions of marker genes in functional categories was used to assess the biological mechanism of fibrosis. RESULTS: The profile data showed several biological changes associated with progression of hepatic fibrosis. Clustered genes in functional categories showed sequential changes in expression. Several sets of clustered genes, including those related to the extracellular matrix (ECM), inflammation, lipid metabolism, steroid metabolism, and some transcription factors important for hepatic biology showed expression changes in the immediate early phase (F1/F2) of fibrosis. Genes associated with aromatic amino acid (AA) metabolism, sulfur-containing AA metabolism and insulin/ Wnt signaling showed expression changes in the middle phase (F2/F3), and some genes related to glucose metabolism showed altered expression in the late phase of fibrosis (F3/F4). Therefore, molecular networks showing serial changes in gene expression are present in liver fibrosis progression in hepatitis C patients. CONCLUSION: Analysis of gene expression profiles from a perspective of functional categories or molecular networks provides an understanding of disease and suggests new diagnostic methods. Selected marker genes have potential utility for biological identification of advanced fibrosis. PMID- 18395901 TI - Pancreatic stellate cells promote proliferation and invasiveness of human pancreatic cancer cells via galectin-3. AB - AIM: To investigate the role of pancreatic stellate cells (PSCs) and galectin-3 (GAL-3) in the proliferation and infiltration of pancreatic cancer cell line SW1990. METHODS: Human pancreatic cancer cell line SW1990 and PSCs were cultured in vitro. Supernatant fluid of cultured PSCs and SW1990 cells was collected. Expression of GAL-3 in SW1990 cells and PSCs was detected by ELISA, RT-PCR and Western blotting. Proliferation of cultured PSCs and SW1990 cells was measured by 3-(4, 5-methylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Infiltration of SW1990 cells was detected by a cell infiltration kit. RESULTS: SW1990 cells expressed GAL-3 and this was up-regulated by the supernatant fluid of cultured PSCs. PSCs did not express GAL-3. SW1990 cells stimulated proliferation of PSCs via GAL-3. GAL-3 antibody inhibited SW1990 cell proliferation, while the supernatant fluid of PSCs stimulated proliferation of SW1990 cells through interaction with GAL-3 protein. The supernatant fluid of PSCs enhanced the invasiveness of SW1990 cells through interaction with GAL-3. CONCLUSION: GAL-3 and PSCs were involved in the proliferation and infiltration process of pancreatic cancer cells. PMID- 18395902 TI - Probiotic intervention has strain-specific anti-inflammatory effects in healthy adults. AB - AIM: To evaluate the effects of three potentially anti-inflammatory probiotic bacteria from three different genera on immune variables in healthy adults in a clinical setting based on previous in vitro characterization of cytokine responses. METHODS: A total of 62 volunteers participated in this randomized, double-blind and placebo-controlled parallel group intervention study. The volunteers were randomized to receive a milk-based drink containing either Lactobacillus rhamnosus GG (LGG), Bifidobacterium animalis ssp. lactis Bb12 (Bb12), or Propionibacterium freudenreichii ssp. shermanii JS (PJS) or a placebo drink for 3 wk. Venous blood and saliva samples were taken at baseline and on d 1, 7 and 21. Fecal samples were collected at baseline and at the end of intervention. RESULTS: The serum hsCRP expressed as the median AUC(0-21) (minus baseline) was 0.018 mg/L in the placebo group, -0.240 mg/L in the LGG group, 0.090 mg/L in the Bb12 group and -0.085 mg/L in the PJS group (P = 0.014). In vitro production of TNF-alpha from in vitro cultured peripheral blood mononuclear cells (PBMC) was significantly lower in subjects receiving LGG vs placebo. IL-2 production from PBMC in the Bb12 group was significantly lower compared with the other groups. CONCLUSION: In conclusion, probiotic bacteria have strain-specific anti-inflammatory effects in healthy adults. PMID- 18395903 TI - A combination therapy of ethanol injection and radiofrequency ablation under general anesthesia for the treatment of hepatocellular carcinoma. AB - AIM: To summarize the effects of laparoscopic ethanol injection and radiofrequency ablation (L-EI-RFA), thoracoscopic (T-EI-RFA) and open-surgery assisted EI-RFA (O-EI-RFA) under general anesthesia for the treatment of hepatocellular carcinoma (HCC). METHODS: Time-lag performance of RFA after ethanol injection (Time-lag PEI-RFA) was performed in all cases. The volume of coagulated necrosis and the applied energy for total and per unit volume coagulated necrosis were examined in the groups treated under general (group G) or local anesthesia (group L). RESULTS: The results showed that the total applied energy and the applied energy per unit volume of whole and marginal, coagulated necrosis were significantly larger in group G than those in the group L, resulting in a larger volume of coagulated necrosis in the group G. The rate of local tumor recurrence within one year was extremely low in group G. CONCLUSION: These results suggest that EI-RFA, under general anesthesia, may be effective for the treatment of HCC because a larger quantity of ethanol and energy could be applied during treatment under painfree condition for the patients. PMID- 18395904 TI - Serum type IV collagen level is predictive for esophageal varices in patients with severe alcoholic disease. AB - AIM: To determine factors predictive for esophageal varices in severe alcoholic disease (SAD). METHODS: Abdominal ultrasonography (US) was performed on 444 patients suffering from alcoholism. Forty-four patients found to have splenomegaly and/ or withering of the right liver lobe were defined as those with SAD. SAD patients were examined by upper gastrointestinal (UGI) endoscopy for the presence of esophageal varices. The existence of esophageal varices was then related to clinical variables. RESULTS: Twenty-five patients (56.8%) had esophageal varices. A univariate analysis revealed a significant difference in age and type IV collagen levels between patients with and without esophageal varices. A logistic regression analysis identified type IV collagen as the only independent variable predictive for esophageal varices (P = 0.017). The area under the curve (AUC) for type IV collagen as determined by the receiver operating characteristic (ROC) for predicting esophageal varices was 0.78. CONCLUSION: This study suggests that the level of type IV collagen has a high diagnostic accuracy for the detection of esophageal varices in SAD. PMID- 18395905 TI - Early effects of Lansoprazole orally disintegrating tablets on intragastric pH in CYP2C19 extensive metabolizers. AB - AIM: To compare rabeprazole (RPZ; 10 mg) with Lansoprazole orally disintegrating tablets (LPZ; 30 mg OD) in terms of antisecretory activity and blood drug concentration after a single dose. METHODS: Eight H pylori -negative cytochrome P450 (CYP) 2C19 extensive metabolizers were assigned to receive a single oral dose of RPZ 10 mg or LPZ 30 mg OD. Twelve hour intragastric pH monitoring was performed on the day of treatment. Blood samples were also collected after the administration of each drug. RESULTS: LPZ 30 mg OD induced a significantly earlier rise in blood drug concentration than RPZ 10 mg; consequently, LPZ 30 mg OD induced a significantly earlier rise in median pH in the third and fourth hours of the study. CONCLUSION: In H pylori-negative CYP2C19 extensive metabolizers, LPZ 30 mg OD induced a significantly faster inhibition of gastric acid secretion than RPZ 10 mg. PMID- 18395906 TI - p16 promoter hypermethylation: a useful serum marker for early detection of gastric cancer. AB - AIM: To determine p16 promoter hypermethylation in gastric tumoral tissue and serum samples, its impact on p16-protein expression, and correlation with clinical and histological features. METHODS: Samples were obtained from 52 histologically confirmed cases of gastric adenocarcinoma. Gastric tissue and serum of 50 age- and sex-matched individuals with normal gastroscopy and biopsy were obtained as control samples. Methylation-specific polymerase chain reaction (MSP) was used to evaluate methylation status of p16 promoter. p16-protein expression was analyzed by immunohistochemical staining on paraffin-embedded sections. RESULTS: Methylation was detected in 44.2% (23/52) of tumoral tissues. 60.9% of them were also methylated in serum, i.e., 26.9% of all patients (14/52). Methylation was not detected in tissue and sera of control samples. p16-protein expression was decreased in 61.5% of cases (32/52), and was significantly associated with promoter hypermethylation (P < 0.001). Methylation was significantly more frequent in higher pathological grades (P < 0.05). Methylation was not associated with other clinicopathological features and environmental factors including H pylori infection and smoking. CONCLUSION: p16 promoter hypermethylation is an important event in gastric carcinogenesis. It is the principle mechanism of p16 gene silencing. It is related to malignant tumor behavior. Detection of DNA methylation in serum may be a biomarker for early detection of gastric cancer. PMID- 18395907 TI - Prospective evaluation of small bowel preparation with bisacodyl and sodium phosphate for capsule endoscopy. AB - AIM: To determine the effect of Prepacol, a combination of sodium phosphate and bisacodyl, on transit and quality of capsule endoscopy (CE). METHODS: Fivety two consecutive patients were included in this prospective study. CE was performed following a 12 h fasting period. Twenty six patients were randomized for additional preparation with Prepacol. The quality of CE was assessed separately for the proximal and the distal small bowel by 3 experienced endoscopists on the basis of a graduation which was initially developed with 20 previous CE. RESULTS: Preparation with Prepacol accelerated small bowel transit time (262 +/- 55 min vs 287 +/- 97 min), but had no effect on the quality of CE. Visibility was significantly reduced in the distal compared to the proximal small bowel. CONCLUSION: The significantly reduced visibility of CE in the distal small bowel allocates the need for a good preparation. Since Prepacol has no beneficial effect on CE the modality of preparation and the ideal time of application remains unclear. Further standardized examinations are necessary to identify sufficient preparation procedures and to determine the impact of the volume of the preparation solution. PMID- 18395908 TI - Effect of Prometheus liver assist system on systemic hemodynamics in patients with cirrhosis: a randomized controlled study. AB - AIM: To evaluate treatment safety and hemodynamic changes during a single 6-h treatment with the Prometheus liver assist system in a randomized, controlled study. METHODS: Twenty-four patients were randomized to either the study group or to one of two control groups: Fractionated Plasma Separation Adsorption and Dialysis, Prometheus system (Study group; n = 8); Molecular Adsorbent Recirculation System (MARS) (Control group 1, n = 8); or hemodialysis (Control group 2; n = 8). All patients included in the study had decompensated cirrhosis at the time of the inclusion into the study. Circulatory changes were monitored with a Swan-Ganz catheter and bilirubin and creatinine were monitored as measures of protein-bound and water-soluble toxins. RESULTS: Systemic hemodynamics did not differ between treatment and control groups apart from an increase in arterial pressure in the MARS group (P = 0.008). No adverse effects were observed in any of the groups. Creatinine levels significantly decreased in the MARS group (P = 0.03) and hemodialysis group (P = 0.04). Platelet count deceased in the Prometheus group (P = 0.04). CONCLUSION: Extra-corporal liver support with Prometheus is proven to be safe in patients with endstage liver disease but does not exert the beneficial effects on arterial pressure as seen in the MARS group. PMID- 18395909 TI - Ultrasonography in differentiation between chronic viral hepatitis and compensated early stage cirrhosis. AB - AIM: To assess the value of gray scale (GS) and colour Doppler ultrasonography (CDU) in differentiating the progression of chronic viral hepatitis (CVH) and compensated liver cirrhosis (CIR). METHODS: Seventy-two patients and 32 normal individuals who were used as controls were studied. Forty-four patients suffered from CVH and 28 from CIR. All patients were underwent to liver biopsy. Multiple qualitative and quantitative variables were studied in liver, portal vein (PV), hepatic artery (HA) and spleen with GS and CDU. On the basis of the obtained CDU data, several known indexes were calculated. In addition, alternative indices [PV diameter (D)/time average mean velocity (VTAM), HA/PV VTAM ratio] were calculated and studied. RESULTS: ROC analysis showed that PV congestion index, PV D/VTAM and HA/PV VTAM indices had the best sensitivity and specificity in discriminating CVH from CIR. Stepwise discriminant analysis showed that 88.9% of the originally grouped cases could be correctly classified by the three qualitative and four quantitative variables selected as statistically significant predictors. Among the CVH patients who underwent to biopsy, statistically significant changes were found in those at fibrosis stage 5 compared to fibrosis stages 1-4. CONCLUSION: Simple GS and CDU parameters discriminate CVH from CIR. The alternative Doppler indexes can accurately differentiate chronic virus hepatitis from cirrhosis. These indexes can be used in monitoring chronic virus hepatitis and avoiding unnecessary biopsies. PMID- 18395910 TI - Endoscopic band ligation and endoscopic hemoclip placement for patients with Mallory-Weiss syndrome and active bleeding. AB - AIM: To compare the hemostatic efficacy and safety of two mechanical endoscopic methods: endoscopic band ligation (EBL) and endoscopic hemoclip placement (EHP) in patients with actively bleeding Mallory-Weiss syndrome (MWS). METHODS: A prospective randomized study to compare the efficacy and safety of EHP with EBL was performed from January 2002 to August 2005. Forty-one patients with active bleeding from MWS were treated with EHP (n = 21) or EBL (n = 20). RESULTS: There were no significant differences between groups with respect to clinical and endoscopic characteristics. The mean number of hemoclips applied was 3.2 +/- 1.5 and the mean number of bands applied was 1.2 +/- 0.4. Primary hemostasis was achieved in all patients. Recurrent bleeding was observed in one patient from the EHP group and two from the EBL group. Patients with recurrent bleeding were treated by the same modality as at randomization and secondary hemostasis was achieved in all. There were no significant differences between the two groups in total transfusion amount or duration of hospital stay. No complications or bleeding-related death resulted. CONCLUSION: EHP and EBL are equally effective and safe for the management of active bleeding in patients with Mallory-Weiss syndrome, even in those with shock or comorbid diseases. PMID- 18395911 TI - Effects of honey as a scolicidal agent on the hepatobiliary system. AB - AIM: To examine the effects of 10% diluted honey, which has been shown to be scolicidal, on the liver and biliary system and determine whether it could be used as a scolicidal agent in the presence of biliary-cystic communication. METHODS: Thirty Wistar-Albino rats were divided into two groups. Honey with 10% dilution in the study group and 0.9% saline (NaCl) in the control group were injected into the common bile ducts of rats through a 3-mm duodenotomy. The animals were sacrificed 6 mo after the procedure. Histopathological, biochemical, and radiological examinations were performed for evaluation of side effects. RESULTS: At the end of the sixth month, liver function tests were found to be normal in both groups. The tissue samples of liver and ductus choledochus of the honey group showed no histomorphologic difference from the control group. No stricture on the biliary tree was detected on the retrograde cholangiograms. CONCLUSION: According to these results, we concluded that 10% diluted honey could be used as scolicidal agent safely in the presence of biliary-cystic communication. PMID- 18395912 TI - Predictive factors for early aspiration in liver abscess. AB - AIM: To determine the predictive factors for early aspiration in liver abscess. METHODS: A retrospective analysis of all patients with liver abscess from 1995 to 2004 was performed. Abscess was diagnosed as amebic in 661 (68%) patients, pyogenic in 200 (21%), indeterminate in 73 (8%) and mixed in 32 (3%). Multiple logistic regression analysis was performed to determine predictive factors for aspiration of liver abscess. RESULTS: A total of 966 patients, 738 (76%) male, mean age 43 +/- 17 years, were evaluated: 540 patients responded to medical therapy while adjunctive percutaneous aspiration was performed in 426 patients. Predictive factors for aspiration of liver abscess were: age > or = 55 years, size of abscess > or = 5 cm, involvement of both lobes of the liver and duration of symptoms > or = 7 d. Hospital stay in the aspiration group was relatively longer than in the non aspiration group. Twelve patients died in the aspiration group and this mortality was not statistically significant when compared to the non aspiration group. CONCLUSION: Patients with advanced age, abscess size > 5 cm, both lobes of the liver involvement and duration of symptoms > 7 d were likely to undergo aspiration of the liver abscess, regardless of etiology. PMID- 18395913 TI - Enhancement of CD4+ T cell activities and modulation of Th1/Th2 lineage development in radiated tumor-bearing rats treated with male zooid of Antheraea pernyi extracts. AB - AIM: To investigate whether supplementation of male zooid of Antheraea pernyi extracts (MZAPE) could enhance immune function of radiated tumor-bearing rats. METHODS: Eighty male Wistar rats were randomly divided into a control group, a simple radiation group, a MZAPE group, and a radiation plus MZAPE group. With the tumor model established by implanting Walker-256 ascites tumor cells, tumor weight and tumor control rate were calculated. The rats in the simple radiation and radiation plus MZAPE groups were underwent to radiation at 10 Gy within 2 d. In the MZAPE and radiation plus MZAPE groups, the MZAPE was gavaged at a dose of 16.53 mg/kg once a day for 7 d. T cell subsets in peripheral blood were determined by flow cytometry and the expression of IL-2, IFN-gamma, IL-4 and IL 10 in sera were determined by ELISA on the 8th d. RESULTS: The tumor weight of simple radiation group, MZAPE group and radiation plus MZAPE group was lower than that of control group (P < 0.01) and tumor control rates were 63.08% +/- 6.43%, 69.86% +/- 7.12% and 35.30% +/- 7.67%, respectively. CD4+ T and CD8+ T cells in the peripheral blood of the simple radiation group were fewer than in control group. In the MZAPE and radiation plus MZAPE groups, the number of CD4+ T cells was higher while CD8+ T cells was lower than in the control and simple radiation groups. Expression of IL-2 and INF-gamma in the radiation group was lower than in control group, and significantly enhanced during MZAPE therapy (P < 0.05). Expression of IL-4 and IL-10 in the radiation group had no significant changes compared with the control group, and decreased significantly after MZAPE treatment (P < 0.01). CONCLUSION: MZAPE administration may help improve the immune function of the radiated tumor-bearing rats and reverse the radiation induced immune inhibition by promoting the proliferation of T helper cells and inducing the transdifferentiation from Th2 to Th1. PMID- 18395914 TI - Effect of Oxymatrine on the TGFbeta-Smad signaling pathway in rats with CCl4 induced hepatic fibrosis. AB - AIM: To explore the anti-fibrotic effect of Oxymatrine on CCl4-induced liver fibrosis in rats and its modulation on the TGFbeta-Smad signaling pathway. METHODS: One hundred healthy male SD rats were randomly divided into three groups: normal group (n = 20), treatment group of Oxymatrine (n = 40) and CCl4 induced fibrosis group (n = 40). Experimental hepatic fibrosis was induced by subcutaneous injection of carbon tetrachloride (CCl4 soluted in liquid paraffin with the concentration of 300 g/L, the dosage of injection was 3 mL/kg, twice per week for 8 wk). The treated rats received Oxymatrine via celiac injection at a dosage of 10 mg/kg twice a week at the same time. The deposition of collagen was observed with H&E and Masson staining. The concentration of serum TGF-beta1 was assayed with ELISA. The gene expression of Smads and CBP (CREB binding protein) was detected with in situ hybridization (ISH) and immunohistochemistry (IH), respectively. All the experimental figures were scanned and analyzed with special figure-analysis software. RESULTS: A significant reduction of collagen deposition and rearrangement of the parenchyma was noted in the liver tissue of Oxymatrine treated rats. The semiquantitative histological scores (2.43 +/- 0.47 microm2 vs 3.76 +/- 0.68 microm2, P < 0.05) and average area of collagen in those rats were significantly decreased when compared with hepatic cirrhosis model rats (94.41 +/ 37.26 microm2 vs 290.86 +/- 89.37 microm2, P < 0.05). The gene expression of Smad 3 mRNA was considerably decreased in the treated animals. The A value of Smad 3 mRNA was lower in the treated rats than the model rats (0.034 +/- 0.090 vs 0.167 +/- 0.092, P < 0.05). Contrarily, the A value of Smad 7 mRNA was increased considerably in the treated animals (0.175 +/- 0.065 vs 0.074 +/- 0.012, P < 0.05). There was an obvious decrease in the expression of CBP mRNA in treated rats as illuminated by a reduction of its A value when compared with model rats (0.065 +/- 0.049 vs 0.235 +/- 0.025, P < 0.001). CONCLUSION: Oxymatrine is effective in reducing the production and deposition of collagen in the liver tissue of experimental rats. Oxymatrine could promote the expression of Smad 7 and inhibit the expression of Smad 3 and CBP in CCl4-induced hepatic fibrosis in SD rats, could modulate the fibrogenic signal transduction of TGFbeta-Smad pathway. PMID- 18395915 TI - Intraperitoneal administration of gonadotropin-releasing hormone-PE40 induces castration in male rats. AB - AIM: To evaluate the long-term effects of gonadotropinreleasing hormone (GnRH) based vaccine on levels of GnRH antibody and testosterone, and vaccine-induced immunocastration on sexual behavior of male rats. METHODS: The rats were treated with GnRH-PE40 intraperitoneally every other day for 12 wk. GnRH antibody and testosterone level in rat blood were determined by ELISA and radioimmunoassay, respectively. Morphological changes in testes and sexual behavior of rats were evaluated. RESULTS: GnRH-PE40 induced a high production in GnRH antibody, decreased the serum testosterone level, testis atrophy and sexual function in rats. CONCLUSION: Intraperitoneal administration of GnRHPE40 produces structural and functional castration of male rat reproductive system by inducing anti-GnRH antibody. PMID- 18395916 TI - Expression of connective tissue growth factor in tumor tissues is an independent predictor of poor prognosis in patients with gastric cancer. AB - AIM: To examine the expression of connective tissue growth factor (CTGF), also known as CCN2, in gastric carcinoma (GC), and the correlation between the expression of CTGF, clinicopathologic features and clinical outcomes of patients with GC. METHODS: One hundred and twenty-two GC patients were included in the present study. All patients were followed up for at least 5 years. Proteins of CTGF were detected using the Powervision two-step immunostaining method. RESULTS: Of the specimens from 122 GC patients analyzed for CTGF expression, 58 (58/122, 47.5%) had a high CTGF expression in cytoplasm of gastric carcinoma cells and 64 (64/122, 52.5%) had a low CTGF expression. Patients with a high CTGF expression showed a higher incidence of lymph node metastasis than those with a low CTGF expression (P = 0.032). Patients with a high CTGF expression had significantly lower 5-year survival rate than those with a low CTGF expression (27.6% vs 46.9%, P = 0.0178), especially those staging I + II + III (35.7% vs 65.2%, P = 0.0027). CONCLUSION: GC patients with an elevated CTGF expression have more lymph node metastases and a shorter survival time. CTGF seems to be an independent prognostic factor for the successful differentiation of high-risk GC patients staging I + II + III. Over-expression of CTGF in human GC cells results in an increased aggressive ability. PMID- 18395917 TI - Endoscopic ultrasonography-guided trucut biopsy for the preoperative diagnosis of peripancreatic Castleman's disease: a case report. AB - Castleman's disease (CD) of the pancreas/peripancreas is extremely rare. The recently introduced, endoscopic ultrasonography (EUS)-guided trucut biopsy (TCB) is a useful diagnostic modality for obtaining tissue samples from peripancreatic lesions. However, its role in diagnosing CD remains unknown. We report a case of localized, peripancreatic, hyaline-vascular CD biopsied using EUS. The pathology results were initially interpreted as an extranodal, marginal-zone B-cell lymphoma. However, polymerase chain reaction (PCR) study for the IgH gene rearrangement revealed a polyclonal pattern. We also reviewed the relevant literature. To our knowledge, this is the first illustrated report on EUS-TCB findings of CD with its pathology results of EUS-TCB mimicked a B-cell lymphoma. PMID- 18395918 TI - Primary rectal signet ring cell carcinoma with peritoneal dissemination and gastric secondaries. AB - Disseminated signet ring cell carcinomas frequently arise from the stomach. However, primaries in the colon and rectum have also been reported. We present a 68 year old lady who presented with a change in her bowel habit. Colonoscopy showed a stenosing rectal tumour at 7 cm to 8 cm from the anal verge. Multiple scattered ulcers were also noted along the entire length of the colon. Biopsy of the lesions revealed signet ring cell adenocarcinoma. Gastroscopy showed multiple nodules with ulceration over several areas of the stomach which were similar in appearance to the colonic lesions. However, no primary tumour of the stomach was seen. Biopsy of the gastric lesions also showed signet ring cell adenocarcinoma. Computed tomography scan of the abdomen and pelvis revealed circumferential tumour at the rectosigmoid junction with possible invasion into the left ischiorectal fossa. The overall picture was that of a primary rectal signet ring cell carcinoma with peritoneal dissemination. The patient was referred for palliative chemotherapy in view of the disseminated disease. In the present report, we discuss this interesting pathological entity and review the role of various histolological techniques in helping to identify the primary tumor. PMID- 18395919 TI - Diagnosis and treatment of Gardner syndrome with gastric polyposis: a case report and review of the literature. AB - Gardner syndrome (GS) is an autosomal dominant disease characterized by the presence of colonic polyposis, osteoma and soft tissue tumors. It is regarded as a clinical subgroup of familial adenomatous polyposis (FAP) and may present at any age from 2 mo to 70 years with a variety of symptoms, either colonic or extracolonic. We present a case of a 23-year-old female patient with GS who presented with gastric polyposis and was successively treated with restorative proctocolectomy in combination with ileal pouch anal anastomosis (RPC/ IPAA), ileostomy, ileostomy closure operation, snare polypectomy during 8 mo. After operation, the patient took oral traditional Chinese medicine pills made of Fructus mume and Bombyx batryticatu for about 6 mo. The innutrition and anaemia of this patient were gradually improved. Gastroscopy showed that the remnant gastric polypi gradually decreased and finally disappeared 19 mo after the first operation. The patient had 2-3 times of solid stool per day at the time we wrote this paper. PMID- 18395921 TI - Role of ABCC2 common variants in intrahepatic cholestasis of pregnancy. AB - The pathogenesis of intrahepatic cholestasis of pregnancy (ICP), a disorder that adversely affects maternal wellbeing and fetal outcome, is unclear. However, multiple factors probably interact along with a genetic predisposition. We would like to add some comments on a paper recently published concerning the role of ABCB11 and ABCC2 polymorphisms in both ICP and contraceptive-induced cholestasis, especially in the light of our recently published findings about a positive association between ICP and ABCC2 common variants. PMID- 18395922 TI - Brain-derived neurotrophic factor-mediated retrograde signaling required for the induction of long-term potentiation at inhibitory synapses of visual cortical pyramidal neurons. AB - High-frequency stimulation (HFS) induces long-term potentiation (LTP) at inhibitory synapses of layer 5 pyramidal neurons in developing rat visual cortex. This LTP requires postsynaptic Ca2+ rise for induction, while the maintenance mechanism is present at the presynaptic site, suggesting presynaptic LTP expression and the necessity of retrograde signaling. We investigated whether the supposed signal is mediated by brain-derived neurotrophic factor (BDNF), which is expressed in pyramidal neurons but not inhibitory interneurons. LTP did not occur when HFS was applied in the presence of the Trk receptor tyrosine kinase inhibitor K252a in the perfusion medium. HFS produced LTP when bath application of K252a was started after HFS or when K252a was loaded into postsynaptic cells. LTP did not occur in the presence of TrkB-IgG scavenging BDNF or function blocking anti-BDNF antibody in the medium. In cells loaded with the Ca2+ chelator BAPTA, the addition of BDNF to the medium enabled HFS to induce LTP without affecting baseline synaptic transmission. These results suggest that BDNF released from postsynaptic cells activates presynaptic TrkB, leading to LTP. Because BDNF, expressed activity dependently, regulates the maturation of cortical inhibition, inhibitory LTP may contribute to this developmental process, and hence experience-dependent functional maturation of visual cortex. PMID- 18395923 TI - Directed immobilization of DNA-binding proteins on a cognate DNA-modified chip surface. AB - Here we describe a useful method for the site-directed immobilization of proteins with a DNA-binding domain (DNA-BD) on the cognate DNA-coated gold surface for surface plasmon resonance (SPR) imaging analyses. In order to assess the performance of this procedure, we utilized two DNA-BDs, yeast GAL4 DNA-BD, and bacterial LexA DNA-BD. After the immobilization of the cognate double-stranded DNAs (dsDNAs) to a gold chip surface with a monolayer of poly(l-lysine) for sequence-specific DNA-protein interaction, purified recombinant GAL4 DNA-BD:EGFP and LexA DNA-BD:RFP fusion proteins were applied to a dsDNA-spotted gold chip, and were subsequently analyzed using an SPR imaging system. Consequently, the recombinant DNA-binding proteins, GAL4 DNA-BD:EGFP and LexA DNA-BD:RFP, were shown to bind selectively to their cognate DNA sequences on the gold chip. Collectively, our results revealed that sequence-specific dsDNA microarray approach could prove useful in performing the site-directed immobilization of DNA binding proteins onto a gold thin film in a parallel format, and thereby potentially allowing for the analysis of transcription factor binding profiling as well as for the monitoring of protein-protein interactions between target proteins with DNA-binding domain as a fusion tag and their binding partners. PMID- 18395924 TI - On-line monitoring of lipid storage in yeasts using impedance spectroscopy. AB - Bioremediation technologies and many environmentally sound biosyntheses rely on the catalytic potential of whole cells. For analyzing and controlling such processes robust real-time indicators for the concentration of intact cells such as impedance are required. The conventional method measures the capacitances of cell suspensions at one or two frequencies and correlates them with biomass concentrations. However, cell inclusions such as lipid droplets or overproduced enzymes may block intracellular ion paths, thereby possibly modifying the dielectric properties of the cells. To test the hypothesis that the total impedance spectrum into the analysis may provide useful information about cell inclusions, the impedance spectrum of a technical culture of the oleaginous yeast Arxula adeninivorans was measured and evaluated every 15s. This yeast is a good test object since it stores the excess of assimilated carbon in experimentally controllable lipid droplets. Upon correction for possible impedance signal interferences, we derived different empirical methods suitable to indicate incipient lipid formation. The methods were designed to act on-line and are thus principally suited for real-time monitoring of cell inclusions. In search for optimised bioprocess monitoring we tested a heuristic spectrum analysis using integrative statistics (RDA). With this approach we were able to accurately detect the formation of cell inclusions, which is potentially valuable for future bioprocess control strategies. PMID- 18395925 TI - In vivo selection of intrabodies specifically targeting protein-protein interactions: a general platform for an "undruggable" class of disease targets. AB - Protein-protein interactions represent a major potential drug target for many human diseases, but these are unanimously considered undruggable with small chemical molecules. We have developed 3-SPLINT, a novel technology for the selection of antibodies that are intrinsically endowed with the ability to interfere with a given protein-protein interaction. The selection procedure exploits the recently described yeast SPLINT libraries of intrabodies, adapting them to a reverse-hybrid system, yielding the selection of recombinant antibodies that are able to disrupt a target protein-protein interaction in vivo. This class of antibodies should therefore perturb an individual protein-protein interaction, without perturbing the scaffolding function of the target protein in that complex, or other protein interactions of that same protein. We provide here a proof of concept of the technology, by the de novo selection of antibodies against two distinct interacting protein pairs: the GABARAP, which interact with the gamma2 subunit of GABA(A) receptor, and the p65 protein dimer, involved in the NF-kappaB-mediated signalling transduction pathway. Intrabodies selected against the latter were functionally validated in cells. Such antibodies, by interfering with the dimerization domain of p65, lead to an activation of the NF kappaB-mediated transcriptional activity, which is normally inhibited by p65 knock-down RNAi. This provides a clear-cut demonstration that interfering with a protein interaction can be functionally very different from physically removing one of the interacting proteins. The 3-SPLINT approach provides a general and finer tool for the functional validation of selected protein interactions in protein networks, and is ideally applied to protein "hubs", displaying multiple distinct interactions. 3-SPLINT will therefore complement RNAi-based approaches, in the toolkit of target validation strategies, and is amenable to the systematic isolation of comprehensive sets of antibodies against most protein-protein interactions of a given protein network. PMID- 18395926 TI - Adsorption of insulin onto infusion sets used in adult intensive care unit and neonatal care settings. AB - INTRODUCTION: Insulin adsorption onto infusion equipment may affect glycaemic control. METHODS: The change in insulin concentration during delivery through tubing employed for adult ICU and neonatal patients was determined using continuous flow UV analysis. RESULTS: Insulin adsorption depended on tubing composition, dimensions and flow rate, being highest for neonatal polyvinylchloride tubing at low flow rates. CONCLUSION: In continuous insulin therapy, we should consider the nature of the infusion set and flow rate. PMID- 18395927 TI - High-normal blood pressure, impaired glucose regulation and metabolic syndrome have variable impact on central artery stiffness. AB - AIMS: This cross-sectional study compared carotid artery stiffness in metabolic syndrome (MS) subjects with high-normal blood pressure (HNBP) and/or impaired glucose regulation. METHODS: Eighty-nine subjects (40M and 49F, 53.5+/-9.3 years) with either HNBP, impaired glucose regulation, or both were studied. The following measurements were included: risk factors for MS (waist circumference, blood pressure (BP), glucose, triglycerides and high-density lipoprotein (HDL) cholesterol), and carotid artery stiffness (Doppler ultrasound). RESULTS: Forty four subjects (20M and 24F) were classified as without MS (MS-) and 45 subjects (20M and 25F) as with MS (MS+). As expected, each component of MS was different between the groups (all p<0.05). Arterial distensibility in MS+ was significantly lower than in MS- (0.152+/-0.010 vs. 0.181+/-0.0091/mmHg x 10(-2), p<0.05), while beta stiffness index was not different between the two groups (12.3+/-1.1 vs. 10.5+/-1.1 AU, p=0.115). Multivariate analysis revealed that carotid artery stiffness was independently associated with resting HR together with age (both p<0.05). CONCLUSIONS: These results show that, in subjects with HNBP and/or impaired glucose regulation: (1) the presence of MS decreases arterial distensibility of the carotid artery but do not increase beta stiffness index and (2) the presence of MS dose not synergistically increase carotid artery stiffness. PMID- 18395928 TI - Systematic review of symptom control and quality of life in studies on chemotherapy for advanced non-small cell lung cancer: how CONSORTed are the data? AB - BACKGROUND: The effect of chemotherapy on survival of patients with advanced NSCLC is modest, therefore patient reported outcomes (PRO's) are of high interest in randomized controlled trials (RCTs). CONSORT (CONsolidated Standards On Reporting Trials) is a quality checklist of 22 items for the conduct and reporting of RCTs. The aim of this report was to analyse to what extent the different RCTs with information on PRO's adhere to the CONSORT statement. METHODS: Systematic review of RCTs using PRO's either as primary or secondary endpoint. Compliance with the (revised) CONSORT statement was checked by 2 independent reviewers by making for each study the simple sum of the 22 CONSORT items, or a weighted score with a maximum rating of 31 points. RESULTS: The median weighted CONSORT score of the different RCTs was 25, with a remarkable difference from 12 till 30. There was no significant change over time, nor difference between academic and commercial studies, but a significant correlation between CONSORT agreement and journal type (P<0.0001). Adherence to CONSORT was similar for studies comparing chemotherapy with best supportive care alone, comparing different first-line chemotherapies with PRO either as primary or secondary endpoint, or studies looking at second-line chemotherapy. Benefit in PRO's was reported in all of these settings. CONCLUSION: The overall adherence of peer-reviewed RCTs to CONSORT is reasonable, with nonetheless major differences between journals, and with no clear sign of change over time. Apart from modest survival differences, benefits in PRO endpoints are present in all categories of studies we analysed. PMID- 18395929 TI - Frequent p16 inactivation by homozygous deletion or methylation is associated with a poor prognosis in Japanese patients with pleural mesothelioma. AB - This study examined the p16 expression status and the P16 gene deletion and methylation status in specimens from Japanese patients with malignant pleural mesothelioma (MPM). Immunohistochemical staining for p16 protein and fluorescence in situ hybridization for the P16 gene were performed using specimens from 30 Japanese patients with primary MPM. The methylation status of the P16 gene was examined in 13 patients whose frozen tumor specimens were available using a methylation-specific PCR assay. Among the 30 patients, the loss of p16 protein expression was observed in 24 patients (80.0%). Twenty-one patients had homozygous deletions, and 9 patients retained the P16 gene. None of the patients with P16 homozygous deletions exhibited p16-positive expression, and 3 patients who retained the P16 gene did not exhibit p16-positive expression. Aberrant P16 methylation was present in two patients with an intact P16 gene but without p16 expression. These results suggest that either a homozygous deletion or methylation is responsible for P16 inactivation. Regarding the prognosis, patients with p16-negative expression had a significantly shorter survival time than those with p16-positive expression (P=0.040). Our study showed that P16 inactivation by homozygous deletions or methylation is a frequent event in Japanese patients with MPMs, relating to poor prognosis. Homozygous deletion is the major cause of P16 inactivation, but methylation also lead to the inactivation of P16 when the P16 alleles are retained. PMID- 18395930 TI - Expression of excision repair cross-complementation group 1 and class III beta tubulin predict survival after chemotherapy for completely resected non-small cell lung cancer. AB - In this study, we examined the expression of excision repair cross complementation group 1 (ERCC1) protein in 90 completely resected lung cancer samples from patients who received adjuvant or neo-adjuvant platinum-based chemotherapy. Epidermal growth factor receptor (EGFR) was also studied in these samples. We also examined class III beta-tubulin protein expression in 50 patients treated with a platinum-based drug plus paclitaxel. Among 90 patients treated with platinum-based chemotherapy, the loss of ERCC1 protein expression was associated with a better prognosis (p=0.0068). The effect of ERCC1 expression on survival was not seen in a separate set of 59 patients who underwent curative resection but did not receive adjuvant chemotherapy. Among 50 patients treated with a platinum-based drug plus paclitaxel, loss of class III beta-tubulin protein expression was also associated with a better prognosis (p=0.0303). When combined, patients with a tumor that was negative for both ERCC1 and class III beta-tubulin had a significantly longer overall survival than those with a tumor that expressed either ERCC1 or class III beta-tubulin (p=0.0230). There was no relationship between the presence of an EGFR mutation and the patients' survival after the platinum-based chemotherapy. In conclusion, we found that the loss of ERCC1 and class III beta-tubulin protein expression were predictors of better survival in patients who received a platinum-based plus taxane chemotherapy. PMID- 18395931 TI - Eating disorder risk behavior in Brazilian adolescents from low socio-economic level. AB - We investigated the prevalence, by gender, age and nutritional status, of eating disorder (ED) risk behavior, using a simplified self-report questionnaire in a probabilistic sample of 561, 12-19-year-old students from public schools in the metropolitan area of Rio de Janeiro, Brazil. Sex- and age-specific body mass index cut-offs were used to assess nutritional status. The prevalence of overweight/obesity was 16.2% and of being underweight was 2.5%; 37.3% of the adolescents studied presented symptoms of binge eating (BE) and 24.7% would go on a strict diet at least once a week, both cases more frequent in females (40.8% vs. 25.3%; 31.2% vs. 10.5%, respectively). Older students were shown to be more susceptible to binge eating and younger students more susceptible to strict dieting. Overweight adolescents were shown to be more susceptible to strict dieting than normal-weight adolescents, regardless of sex and age. The prevalence of binge eating and strict dieting was high in low-income Brazilian adolescents and females are at greater risk of developing eating disorders than males. The greater prevalence of strict dieting in younger students shows they are at nutritional risk. PMID- 18395932 TI - Children's nutrition and health in adulthood. AB - Scientific and medical studies report that the environment, and particularly physical activity and the diet, changes the metabolic programming. A systematic review of size and growth during infancy supports conclusions with regard to preventing later obesity. Moreover, the correlation has been established between birth weight, arterial blood pressure, plasmatic lipid level, and cardiovascular mortality. The European Nutrition Committee has written new recommendations with lower protein levels. A lot of data support the application of blocking sensibilisation through intervention in pregnancy and lactation. Primary prevention is possible with breastfeeding for the first 4-6 months of life, particularly for high-risk allergic families. This is a priority for the national health program in France for controlling the increasing obesity. PMID- 18395933 TI - Food memory and its relation with age and liking: an incidental learning experiment with children, young and elderly people. AB - The present study compared incidental learning and food memory in children, young adults and elderly people for three sensory modalities (taste, texture and aroma). The relation of gender and liker-status (i.e. how much we like a product) with food memory was also investigated. Participants received a complete meal including a custard dessert used as target under incidental learning conditions. 24h later, participants were confronted with a series of samples consisting of the target and slightly modified versions of the target (distractors) and were unexpectedly asked to perform an "absolute memory" ("Did you eat this sample yesterday?") and a "relative memory" test ("Is the present sample less/equal/more pleasant than the one you ate yesterday?"). Participants also performed a hedonic and a discrimination test. Memory for the custard was poor and did not depend on age, but it was related to gender, and to how much participants liked the product. Females and high-likers outperformed males and low-likers in the absolute memory task, but they were not better in discriminating the products on both the hedonic and the perceptual dimension. Results also showed that, contrary to common belief, not all sensory aspects that can be discriminated in perception and in liking, are equally well remembered. PMID- 18395934 TI - Warm lidocaine/tetracaine patch versus placebo before pediatric intravenous cannulation: a randomized controlled trial. AB - STUDY OBJECTIVE: We compare the pain of intravenous (IV) cannulation in pediatric emergency department (ED) patients after applying a topical lidocaine/tetracaine patch versus placebo. We hypothesized that application of the active patch would reduce the pain of IV cannulation by at least 15 mm. METHODS: We conducted a randomized, double-blind, placebo-controlled trial in a suburban academic ED. Patients aged 3 to 17 years who required nonemergency IV cannulation were eligible for enrollment. At triage, a nurse placed a commercially available topical lidocaine/tetracaine patch or an identical-looking placebo patch over the antecubital or hand vein in patients for whom an IV catheter was anticipated. After IV cannulation by the treating nurse, the pain of cannulation was measured on a validated 100-mm visual analogue scale or Wong Baker scale. Outcomes were compared between groups with Mann-Whitney U, Student t, and chi(2) tests. A sample of 40 patients had 80% power to detect a 13-mm difference in pain scores. RESULTS: Forty-five patients were randomized to lidocaine/tetracaine patch (22) or placebo (23), and IV cannulation was attempted in 40 of these patients. Mean age was 10 years (SD=4.3), 35% were female patients. The median pain of IV cannulation in the active treatment group (18 mm [interquartile range (IQR) 1 to 40 mm]) was significantly lower than in the placebo group (35 mm [IQR 20 to 59 mm]; P=.04). Adequate pain relief was more common in the active treatment group (75% [95% confidence interval (CI) 53% to 89%] versus 35% [95% CI 18% to 57%]; difference 40% [95% CI 6% to 64%]). The number of successful IV cannulations after the first attempt was similar in both the lidocaine/tetracaine and the placebo groups (90% [95% CI 70% to 97%] versus 85% [95% CI 64% to 95%]; difference 5% (95% CI -21% to 30%). CONCLUSION: Application of a topical lidocaine/tetracaine patch resulted in a modest reduction in the pain of IV cannulation in pediatric ED patients and did not alter the rate of successful cannulations. PMID- 18395935 TI - Characteristics of women with fetal loss in symptomatic first trimester pregnancies with documented fetal cardiac activity. AB - STUDY OBJECTIVE: The purpose of this study is to determine fetal outcomes of women diagnosed with live intrauterine pregnancy after emergency department (ED) presentation for abdominopelvic pain or vaginal bleeding during the first trimester. METHODS: A retrospective medical record review of prospectively recorded data of consecutive ED charts from December 2005 to June 2006 was performed to identify patients diagnosed with live intrauterine pregnancy. Demographic data, obstetric/gynecologic history, and presenting symptoms were obtained. Outcomes were determined by computerized medical records. Fetal loss was diagnosed by decreasing beta-Human chorionic gonadotropin or pathology specimen. Live birth was diagnosed by viable fetus at 20-week-gestation ultrasonography or delivery. RESULTS: A total of 837 patients were evaluated during the first trimester. Three hundred forty patients (41%) met inclusion criteria, with a diagnosis of live intrauterine pregnancy. Outcome data were obtained for 303 (89%) of these patients. Fetal loss occurred in 28 (9.2%) pregnancies (95% confidence interval [CI] 5.9% to 12.5%). Fetal loss incidence was 13.8% (95% CI 9.9% to 17.7%) in patients presenting with vaginal bleeding compared with 2.5% (95% CI 0.007% to 4.3%) in patients without bleeding (P<0.002). Twenty five of 28 (89%) patients with resulting fetal loss presented with vaginal bleeding. Vaginal bleeding was the most important predictor of fetal loss; risk ratio 5.6 (95% CI 1.7 to 18.2). CONCLUSION: Fetal loss before 20 weeks occurs in 9.2% of patients with live intrauterine pregnancy diagnosed by ultrasonography. Vaginal bleeding carries a higher fetal loss rate of 13.8%. These data will assist the emergency physician in counseling women experiencing symptomatic first trimester pregnancy. PMID- 18395936 TI - Career satisfaction in emergency medicine: the ABEM Longitudinal Study of Emergency Physicians. AB - STUDY OBJECTIVE: The primary objective of this study is to measure career satisfaction among emergency physicians participating in the 1994, 1999, and 2004 American Board of Emergency Medicine Longitudinal Study of Emergency Physicians. The secondary objectives are to determine factors associated with high and low career satisfaction and burnout. METHODS: This was a secondary analysis of a cohort database created with stratified, random sampling of 1,008 emergency physicians collected in 1994, 1999, and 2004. The survey consisted of 25 questions on professional interests, attitudes, and goals; 17 questions on training, certification, and licensing; 36 questions on professional experience; 4 questions on well-being and leisure activities; and 8 questions about demographics. Data were analyzed with a descriptive statistics and panel series regression modeling (Stata/SE 9.2 for Windows). Questions relating to satisfaction were scored with a 5-point Likert-like scale, with 1=not satisfied and 5=very satisfied. Questions relating to stress and burnout were scored with a 5-point Likert-like scale, with 1=not a problem and 5=serious problem. During analysis, answers to the questions "Overall, how satisfied are you with your career in emergency medicine?" "How much of a problem is stress in your day-to day work for pay?" "How much of a problem is burnout in your day-to-day work for pay?" were further dichotomized to high levels (4, 5) and low levels (1, 2). RESULTS: Response rates from the original cohort were 94% (945) in 1994, 82% (823) in 1999, and 76% (771) in 2004. In 2004, 65.2% of emergency physicians reported high career satisfaction (4, 5), whereas 12.7% of emergency physicians reported low career satisfaction (1, 2). The majority of respondents (77.4% in 1994, 80.6% in 1999, 77.4% in 2004) stated that emergency medicine has met or exceeded their career expectations. Despite overall high levels of career satisfaction, one-third of respondents (33.4% in 1994, 31.3% in 1999, 31% in 2004) reported that burnout was a significant problem. CONCLUSION: Overall, more than half of emergency physicians reported high levels of career satisfaction. Although career satisfaction has remained high among emergency physicians, concern about burnout is substantial. PMID- 18395937 TI - Ghrelin improves burn-induced multiple organ injury by depressing neutrophil infiltration and the release of pro-inflammatory cytokines. AB - Mechanisms of burn-induced skin and remote organ injury involve oxidant generation and the release of pro-inflammatory cytokines. In this study the possible antioxidant and anti-inflammatory effects of ghrelin were evaluated in a rat model of thermal trauma. Wistar albino rats were exposed to 90 degrees C bath for 10 s to induce thermal trauma. Ghrelin, was administered subcutaneously (10 ng/kg/day) after the burn injury and repeated twice daily. Rats were decapitated at 6 h and 48 h after burn injury and blood was collected for the analysis of pro inflammatory cytokines (TNF-alpha and IL-1beta), lactate dehydrogenase (LDH) activity and antioxidant capacity (AOC). In skin, lung and stomach tissue samples malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) and Na(+)-K(+)-ATPase activity were measured in addition to the histological analysis. DNA fragmentation ratio in the gastric mucosa was also evaluated. Burn injury caused significant increase in both cytokine levels, and LDH activity, while plasma AOC was found to be depleted after thermal trauma. On the other hand, in tissue samples the raised MDA levels, MPO activity and reduced GSH levels, Na(+)-K(+)-ATPase activity due to burn injury were found at control levels in ghrelin-treated groups, while DNA fragmentation in the gastric tissue was also reduced. According to the findings of the present study, ghrelin possesses a neutrophil-dependent anti-inflammatory effect that prevents burn induced damage in skin and remote organs and protects against oxidative organ damage. PMID- 18395939 TI - Disturbance of automatic auditory change detection in dementia associated with Parkinson's disease: A mismatch negativity study. AB - OBJECTIVE: To investigate whether automatic auditory change detection, as measured by the mismatch negativity (MMN) event-related potential waveform, differs in dementia associated with Parkinson's disease (PDD) and dementia with Lewy-bodies (DLB) as compared to Alzheimer's disease (AD), Parkinson's disease without dementia (PD) and healthy control subjects (HC). METHOD: Seventeen DLB, 15 PDD, 16 PD, 16 AD patients and 18 HC subjects participated. A passive MMN event-related potential paradigm and an oddball-distractor reaction time paradigm were presented. RESULTS: The PDD patients had reduced MMN area and amplitude compared to the DLB, PD, and the HC groups. The MMN area correlated significantly with number of missed target stimuli in the oddball-distractor task, and the PDD group missed targets significantly more often than the DLB group. CONCLUSION: The results indicate that PDD patients to a larger degree than patients with DLB have a deficit of automatic auditory change detection that contributes to impairment in their ability to selectively attend and respond to deviant auditory stimuli. PMID- 18395938 TI - Pharmacologic study of C-terminal fragments of frog skin calcitonin gene-related peptide. AB - The calcitonin gene-related peptide from the skin of the frog Phyllomedusa bicolor (pbCGRP) is a 37-residue neuropeptide that differs from human alpha CGRP (halphaCGRP) at 16 positions. The affinities of the C-terminal fragments of pbCGRP and halphaCGRP were evaluated in SK-N-MC cells: pbCGRP(8-37) (K(i)=0.2nM) and pbCGRP(27-37) (K(i)=95nM) were, respectively, 3 times and 20 times more potent than the human fragments halphaCGRP(8-37) and halphaCGRP(27-37). Their antagonistic potencies were measured in SK-N-MC and Col 29 cells, and the rat vas deferens. pbCGRP(8-37) inhibited the halphaCGRP-stimulated production of cAMP by SK-N-MC and Col 29 cells 3 to 4 times more strongly than halphaCGRP(8-37). Thus pbCGRP(8-37) is the most potent CGRP-1 competitive antagonist of all the natural sequences reported to date. pbCGRP(27-37) was also as potent as [D(31), A(34), F(35)] halphaCGRP(27-37), a prototypic antagonist analog derived from structure activity relationship studies of halphaCGRP(8-37). PMID- 18395940 TI - Neurovascular coupling in Alzheimer patients: effect of acetylcholine-esterase inhibitors. AB - Dualistic effects of acetylcholine-esterase inhibitors on neuronal as well as vasoregulative function have been debated. This study investigated for the first time effects of medication on both components. Visually evoked potentials and resultant hemodynamic responses were assessed in Alzheimer patients (n=31) without vascular lesions in a MRI scan and compared to controls (n=20). After baseline recordings (AD0) tests were repeated under 2x1.5 to 2x3mg (AD1) and 2x4.5 to 2x6mg (AD2) rivastigmine/d. Long-term effects were investigated under 6 months of medication (AD2L). The ADAS, MMSE and DEMTECT were used to assess cognitive function at AD0, AD2 and AD2L. Improvement in vasoregulative function was independent from changes in evoked potentials. Acetylcholine-esterase inhibitors demonstrate substantial vascular effects in humans, which are independent from changes in neuronal function. PMID- 18395941 TI - Quantitative structure-activity relationship (QSAR) of aryl alkenyl amides/imines for bacterial efflux pump inhibitors. AB - A quantitative structure-activity relationship (QSAR) analysis has been performed on a data set of 42 aryl alkenyl amides/imines as bacterial efflux pump inhibitors. Several types of descriptors including topological, spatial, thermodynamic, information content and E-state indices have been used to derive a quantitative relationship between the efflux pump inhibiting activity and structural properties. Algorithm based on genetic function approximation method of variable selection was used to generate the model. Statistically significant model (with r(2)=0.87) was obtained with the descriptors like radius of gyration and heat of formation besides E-state indices, AlogP atom types and solvent accessible charged surface area playing an important role in determining the activity of the compounds against bacterial efflux pump. The model was also tested successfully for external validation criteria. The model is not only able to predict the activity of new compounds but also explained the important regions in the molecules in quantitative manner. PMID- 18395942 TI - Synthesis, spectral characterization, in vitro antibacterial, antifungal and cytotoxic activities of Co(II), Ni(II) and Cu(II) complexes with 1,2,4-triazole Schiff bases. AB - A series of metal complexes of cobalt(II), nickel(II) and copper(II) have been synthesized with newly synthesized biologically active 1,2,4-triazole Schiff bases derived from the condensation of 3-substituted-4-amino-5-mercapto-1,2,4 triazole and 8-formyl-7-hydroxy-4-methylcoumarin, which have been characterized by elemental analyses, spectroscopic measurements (IR, UV-vis, fluorescence, ESR), magnetic measurements and thermal studies. Electrochemical study of the complexes is also reported. All the complexes are soluble to limited extent in common organic solvents but soluble to larger extent in DMF and DMSO and are non electrolytes in DMF and DMSO. All these Schiff bases and their complexes have also been screened for their antibacterial (Escherichia coli, Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa and Salmonella typhi) and antifungal activities (Aspergillus niger, Aspergillus flavus and Cladosporium) by MIC method. The brine shrimp bioassay was also carried out to study their in vitro cytotoxic properties. PMID- 18395943 TI - [Evaluation of in-hospital prescription of fluoroquinolones]. AB - PURPOSE: Quinolones are widely prescribed in various hospital departments (about 20% of antibiotics prescriptions in some of them) and antibiotic resistances is increasingly documented. The aim of this prospective study was to assess the prescriptions of quinolones in several departments of medicine of Rouen University Hospital and compare them to the guidelines of good medical practice for antibiotic prescriptions. METHODS: This study was performed in four medicine departments during two months. When a physician prescribed a quinolone treatment, he had to record informations regarding the infection (type and site, bacteriological proof or not), the patient (age, sex and glomerular filtration rate) and the prescription of quinolone (indication, dosage, administration, combination with another antibiotic, duration of treatment). RESULTS: A hundred and three prescriptions of quinolones were analysed (mean age 75+/-0.5 years). Quinolones treatments were more often used alone, in first intention and without bacteriological proof. The dosage of quinolones was generally adapted but the duration of treatment was often excessive. Quinolones prescriptions were in majority in accordance with the guidelines for pulmonary infections, but were more often inappropriate for urinary infections. CONCLUSION: According to the infections, this study shows discrepancies with the published guidelines with respect to prescription of quinolones, but there are also differences between available guidelines that may at least in part explain our results. An harmonization and a large diffusion of uniform guidelines could improve patients antibiotherapy. PMID- 18395944 TI - [Rabeprazole induced acute interstitial nephritis]. AB - A 84-year-old women underwent renal biopsy because of rapidly progressive renal failure. Rabeprazole induced interstitial nephritis was diagnosed. Interstitial nephritis may complicate the course of any proton pump inhibitor treatment. It is a rare and serious complication. Clinician's awareness of this adverse event is essential for early diagnosis and prompt recovery. PMID- 18395945 TI - [Therapeutic consequences of thrombophilic testing]. AB - OBJECTIVE: The objectives of this article are to review the data about the consequences of thrombophilia testing and to think about its indications. CURRENT KNOWLEDGE AND KEY POINTS: The indications of congenital thrombophilic testing have extended since the discovery of prevalent abnormalities, such as mutations of factor V or II genes. However, thrombophilia does not result in a significant increase in the risk of recurrence unlike the spontaneous occurrence of thrombotic events. The factor V Leiden mutation is associated with a moderate increase in recurrence rate, while the G20210A mutation of factor II is not associated with a significant increase in recurrence. Regarding the decrease in natural anticoagulants is concerned, there is no definite conclusion, although the decrease in antithrombin is suspected of being associated with an increase in recurrence. FUTURE PROSPECTS AND PROJECTS: Finally, identification of a constitutional thrombophilia most often do not influence the therapeutic decisions unless some rare abnormalities are found, such as a decrease in antithrombin, homozygous mutations in factors V or II genes or associations of thrombophilia. One must remember that antiphospholipid antibodies must be searched because their impact on recurrences is well-known. Diagnostic work-up for thrombophilia is not useful after a distal or a superficial venous thrombosis (except for antiphospholipid antibodies in case of distal venous thrombosis). PMID- 18395946 TI - Screening malnutrition in hospital outpatients. Can the SNAQ malnutrition screening tool also be applied to this population? AB - BACKGROUND & AIMS: It is known from earlier studies that only 15% of the malnourished hospital outpatient population is recognized and receives nutritional treatment. To increase this number, a quick and easy malnutrition screening tool would be helpful. Because such a tool is lacking, we developed one by using the SNAQ (Short Nutritional Assessment Questionnaire) as a basis. The aim of this study was to develop a quick and easy malnutrition screening tool and to measure its diagnostic accuracy in malnourished hospital outpatients. METHODS: First, an optimal set of questions was selected for the preoperative outpatient population. Secondly, the diagnostic accuracy for the preoperative outpatients was determined (979 patients) and finally, the diagnostic accuracy for general hospital outpatients was established (705 patients). RESULTS: The three original SNAQ questions proved to be the best set of questions for the outpatient population as well. In the preoperative and general outpatient population the diagnostic accuracy resulted respectively in a sensitivity of 53% and 67%, a specificity of 97% and 98%, a positive predictive value of 69% and 72% and a negative predictive value of 94% and 97%. CONCLUSIONS: With an acceptable diagnostic accuracy it may be concluded that the original SNAQ malnutrition screening tool is valid for the hospital outpatient population. PMID- 18395947 TI - Level of protection of chickens against highly pathogenic H5 avian influenza virus with Newcastle disease virus based live attenuated vector vaccine depends on homology of H5 sequence between vaccine and challenge virus. AB - Vaccination of poultry against avian influenza is of high priority, in particular after the dramatic spread of subtype H5N1 in Asia, Africa and Europe. Newcastle disease virus (NDV) has been developed as a vector for the expression of the main immunogen of avian influenza virus, hemagglutinin (HA). An NDV vector based vaccine has several advantages. It allows easy serological differentiation between infected and vaccinated animals by the detection of antibodies against non-HA influenza proteins. Moreover, it can be administered easily to large numbers of animals by spray or drinking water. We recently showed that chickens could be protected against infection with highly pathogenic avian influenza virus (HPAIV) A/chicken/Italy/8/98 (H5N2) after immunization with a recombinant Newcastle disease virus, NDVH5m, which expresses the homologous hemagglutinin. Here, we describe that immunization with NDVH5m conferred only partial protection against lethal infection with heterologous HPAIV A/duck/Vietnam/TG24-01/05 (H5N1). Comparison of amino acid sequences of both H5 proteins showed only 93.6% amino acid identity. Therefore, a new NDV recombinant (NDVH5Vm) was generated which expresses the H5 protein of HPAIV A/chicken/Vietnam/P41/05 (H5N1). This recombinant virus protected chickens against lethal infection with HPAIV H5N1 (Vietnam) already after one immunization. Our data thus show that application of a vector-based vaccine in the control of influenza may require adaptation of the vaccine to currently circulating viruses. PMID- 18395948 TI - Despite Leishvaccine and Leishmune trigger distinct immune profiles, their ability to activate phagocytes and CD8+ T-cells support their high-quality immunogenic potential against canine visceral leishmaniasis. AB - Phenotypic features of peripheral blood leukocytes have been investigated as a pre-requisite to characterize the protective immunity attributed to both Leishvaccine and Leishmune. Our results showed that either those vaccine were accompanied by distinct profiles on innate immune compartment. While Leishvaccine promoted early changes in phenotypic features of neutrophils and eosinophils with late involvement of monocytes, Leishmune induced early and persistent activation of neutrophils and monocytes, without changes on eosinophil activation status. Regarding the adaptive immunity, Leishvaccine sponsored a mixed profile, associated with phenotypic changes of T and B-lymphocytes. Major phenotypic changes in CD4+ T-cells with transient activation of CD8+ T-cell, besides decreased frequency of B-cell expressing CD32 were the hallmark of Leishvaccine. In contrast, Leishmune was associated with phenotypic changes in T-lymphocytes, particularly in CD8+ T-cells, and selective up-regulation of CD3+CD5+LowCD8+ cells. We hypothesized that this dissimilar alteration in immunological events would represent phenomenon directly related with the molecular nature of these vaccines besides the distinct adjuvants employed. However, it is important to emphasize that both immunobiologicals are able to activate phagocytes and CD8+ T cells and therefore could be considered priority vaccines with a high-quality immunogenic potential against CVL. PMID- 18395949 TI - Cosmogenic 22Na as an index of the residence time of water in freshwater basins: a review. AB - The goal of this review is to draw attention to an opportunity for the use of cosmogenic (22)Na for dating young surface and underground waters. After 1961 when a significant quantity of (22)Na was released into the environment as a result of nuclear weapon tests, its concentrations in river waters were greatly increased, and a return to natural (cosmogenic) levels took until the mid-1980s. The studies made during this non-steady-state period showed that the one-box model for freshwater basin correctly describes the experimental data. For the 19 studied basins of Russia, the Baltic States and Japan, a calculation based on this model gave values for the mean residence time of water in the range from 4 to 23 years. Now, only cosmogenic (22)Na is in the environment, and it is the single cosmogenic radionuclide at present, which can serve as a steady-state tracer for dating young waters (up to some decades). PMID- 18395950 TI - Using your computer monitor as an X-ray viewing box. PMID- 18395951 TI - EMED Scientific Meeting ESM. PMID- 18395952 TI - Univariate and multivariate characterization of heavy fuel oil weathering and biodegradation in soil. AB - Polycyclic aromatic compounds (PACs) of petrogenic origin are widespread contaminants of the environment. We investigated the weathering and biodegradation of fuel oil PACs in microcosms that contained natural soil or natural soil primed by adding bioremediated soil to a 2% concentration. In the primed soil, lag-phases were shorter, and biodegradation was more pronounced. To describe PAC degradation, changes in the relative distribution of PACs and changes in isomer ratios, that are specific for biodegradation, were determined by univariate and multivariate techniques. A multivariate approach based on normalized areas or isomer ratios, in combination with principal component analysis, provided the most detailed description of weathering and biodegradation. In particular, the sequence of degradation of PACs was more transparent when using the multivariate approach. We also identified new diagnostic ratios of dimethylphenanthrenes for use in the study of moderately and severely biodegraded samples. PMID- 18395953 TI - Foreword: Biomonitoring Equivalents special issue. AB - The challenge of interpreting results of biomonitoring for environmental chemicals in humans is highlighted in this Foreword to the Biomonitoring Equivalents (BEs) special issue of Regulatory Toxicology and Pharmacology. There is a pressing need to develop risk-based tools in order to empower scientists and health professionals to interpret and communicate the significance of human biomonitoring data. The BE approach, which integrates dosimetry and risk assessment methods, represents an important advancement on the path toward achieving this objective. The articles in this issue, developed as a result of an expert panel meeting, present guidelines for derivation of BEs, guidelines for communication using BEs and several case studies illustrating application of the BE approach for specific substances. PMID- 18395954 TI - WITHDRAWN: Ethical issues and ethics reviews in social science research. AB - This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause. PMID- 18395955 TI - Exercise, APOE, and working memory: MEG and behavioral evidence for benefit of exercise in epsilon4 carriers. AB - Performance on the Sternberg working memory task, and MEG cortical response on a variation of the Sternberg task were examined in middle-aged carriers and non carriers of the APOE epsilon4 allele. Physical activity was also assessed to examine whether exercise level modifies the relationship between APOE genotype and neurocognitive function. Regression revealed that high physical activity was associated with faster RT in the six- and eight-letter conditions of the Sternberg in epsilon4 carriers, but not in the non-carriers after controlling for age, gender, and education (N=54). Furthermore, the MEG analysis revealed that sedentary epsilon4 carriers exhibited lower right temporal lobe activation on matching probe trials relative to high-active epsilon4 carriers, while physical activity did not distinguish non-carriers (N=23). The M170 peak was identified as a potential marker for pre-clinical decline as epsilon4 carriers exhibited longer M170 latency, and highly physically active participants exhibited greater M170 amplitude to matching probe trials. PMID- 18395956 TI - Elevated phosphatidylinositol 3-kinase activation and its clinicopathological significance in cervical cancer. AB - OBJECTIVE: The objective was to study the role of PI3K signaling in the development of cervical cancer and the antitumor effect of PI3K inhibitors. STUDY DESIGN: PI3K protein and mRNA expression of cervical cancer and non-neoplastic tissues were analyzed by Western blotting and RT-PCR. PI3K/Akt/mTOR pathway components in HeLa cells were assessed by immunocytochemistry and Western blotting. The inhibitive effect of LY294002 on HeLa cells was studied using MTT assay and flow cytometry. RESULTS: PI3K protein expression was detected in 25 out of 31 tumor specimens. Compared with non-neoplastic tissues, significant overexpression was observed in tumor tissues. For PI3K overexpression with all clinicopathological features, a decreasing trend in adenocarcinoma, advanced stage, and grade was observed. PI3K inhibitor LY294002 efficiently inhibited HeLa cell growth with IC50 of 20.77 microM, and induced apoptosis. The apoptotic rate was 36% at 3h after LY294002 treatment. These pharmacological roles of LY294002 might be played through the PI3K/Akt/mTOR signaling pathway. CONCLUSIONS: The PI3K signaling pathway was implicated in the development of cervical cancer. The activation of its signaling molecules might have clinical implications. Novel targeted therapies for the PI3K/Akt/mTOR signaling pathway components could provide a useful adjuvant therapeutic strategy for cervical cancer. PMID- 18395957 TI - The effect of tobacco exposure on maternal and fetal thyroid function. AB - OBJECTIVE: To determine the effect of smoking on maternal and fetal thyroid function. STUDY DESIGN: This prospective cohort study involved healthy women undergoing elective cesarean section for term singleton infants. Maternal and fetal thyroid indices were compared between smokers (n=21) and non-smokers (n=83). RESULTS: Maternal thyrotropin (TSH) concentrations were significantly lower in smokers than non-smokers (1.75 mIU/L versus 2.15 mIU/L, respectively, p=0.007), with similar free thyroxine (FT4) concentrations (9.59 pmol/L versus 9.56 pmol/L, p=0.755). For women who smoked, the correlation between the average number of cigarettes smoked per day and maternal TSH concentrations was -0.427, p=0.054. Infants of smokers and non-smokers had similar umbilical TSH (5.43 mIU/L versus 5.82 mIU/L, p=0.124) and FT4 concentrations (13.06 pmol/L versus 13.57 pmol/L, p=0.049). CONCLUSION: We demonstrated for the first time that women who smoke during pregnancy have significantly lower TSH concentrations than non smokers. PMID- 18395958 TI - Primary rhabdomyosarcoma with an epithelioid appearance of the fallopian tube: an adult case. PMID- 18395959 TI - Roles of two surface residues near the access channel in the substrate recognition by cytochrome P450cam. AB - Detailed stopped-flow kinetics of binding of 1R-camphor to cytochrome P450cam has been studied at different temperatures for the wild type as well as for two site specific mutants T192E and S190D of the enzyme, where the surface exposed Threonine and Serine residues were mutated by acidic amino acids. The near-UV and visible circular dichroism spectra as well as the intrinsic fluorescence spectra of the WT and mutant enzymes showed that the mutation of the enzyme did not affect the tertiary structure of the enzyme over the temperature range 4-30 degrees C. The S190D mutation did not show any significant change in the rate constants of the substrate association while they were much lower in the T192E mutant compared to the WT enzyme. The activation energies for substrate association and dissociation processes were determined from the analysis of temperature dependence of the rate constants by the Arrhenius equation over the temperature range 4-19 degrees C. The activation energy for the substrate association was found to be significantly higher in the T192E mutant compared to the S190D mutant or the WT enzyme. The results showed that the Threonine 192 that resides on the F-G loop and directed towards the putative substrate access channel of the enzyme, plays an important role in recognition of the substrate at the surface of the enzyme. These results showed that though the active site of the enzyme resides deep inside the protein matrix, the substrate is recognized at the surface of the enzyme and directed towards the active site through the access channel. PMID- 18395960 TI - Building spectral libraries for wetlands land cover classification and hyperspectral remote sensing. AB - Recent advances in remote sensing provide opportunities to map plant species and vegetation within wetlands at management relevant scales and resolutions. Hyperspectral imagers, currently available on airborne platforms, provide increased spectral resolution over existing space-based sensors that can document detailed information on the distribution of vegetation community types, and sometimes species. Development of spectral libraries of wetland species is a key component needed to facilitate advanced analytical techniques to monitor wetlands. Canopy and leaf spectra at five sites in California, Texas, and Mississippi were sampled to create a common spectral library for mapping wetlands from remotely sensed data. An extensive library of spectra (n=1336) for coastal wetland communities, across a range of bioclimatic, edaphic, and disturbance conditions were measured. The wetland spectral libraries were used to classify and delineate vegetation at a separate location, the Pacheco Creek wetland in the Sacramento Delta, California, using a PROBE-1 airborne hyperspectral data set (5m pixel resolution, 128 bands). This study discusses sampling and collection methodologies for building libraries, and illustrates the potential of advanced sensors to map wetland composition. The importance of developing comprehensive wetland spectral libraries, across diverse ecosystems is highlighted. In tandem with improved analytical tools these libraries provide a physical basis for interpretation that is less subject to conditions of specific data sets. To facilitate a global approach to the application of hyperspectral imagers to mapping wetlands, we suggest that criteria for and compilation of wetland spectral libraries should proceed today in anticipation of the wider availability and eventual space-based deployment of advanced hyperspectral high spatial resolution sensors. PMID- 18395961 TI - Quantitative assessment of breast lesion viscoelasticity: initial clinical results using supersonic shear imaging. AB - This paper presents an initial clinical evaluation of in vivo elastography for breast lesion imaging using the concept of supersonic shear imaging. This technique is based on the combination of a radiation force induced in tissue by an ultrasonic beam and an ultrafast imaging sequence capable of catching in real time the propagation of the resulting shear waves. The local shear wave velocity is recovered using a time-offlight technique and enables the 2-D mapping of shear elasticity. This imaging modality is implemented on a conventional linear probe driven by a dedicated ultrafast echographic device. Consequently, it can be performed during a standard echographic examination. The clinical investigation was performed on 15 patients, which corresponded to 15 lesions (4 cases BI-RADS 3, 7 cases BI-RADS 4 and 4 cases BI-RADS 5). The ability of the supersonic shear imaging technique to provide a quantitative and local estimation of the shear modulus of abnormalities with a millimetric resolution is illustrated on several malignant (invasive ductal and lobular carcinoma) and benign cases (fibrocystic changes and viscous cysts). In the investigated cases, malignant lesions were found to be significantly different from benign solid lesions with respect to their elasticity values. Cystic lesions have shown no shear wave propagate at all in the lesion (because shear waves do not propage in liquid). These preliminary clinical results directly demonstrate the clinical feasibility of this new elastography technique in providing quantitative assessment of relative stiffness of breast tissues. This technique of evaluating tissue elasticity gives valuable information that is complementary to the B-mode morphologic information. More extensive studies are necessary to validate the assumption that this new mode potentially helps the physician in both false-positive and false-negative rejection. PMID- 18395962 TI - Transmitted ultrasound pressure variation in micro blood vessel phantoms. AB - Silica, cellulose and polymethylmethacrylate tubes with inner diameters of ten to a few hundred microns are commonly used as blood vessel phantoms in in vitro studies of microbubble or nanodroplet behavior during insonation. However, a detailed investigation of the ultrasonic fields within these micro-tubes has not yet been performed. This work provides a theoretical analysis of the ultrasonic fields within micro-tubes. Numerical results show that for the same tube material, the interaction between the micro-tube and megaHertz-frequency ultrasound may vary drastically with incident frequency, tube diameter and wall thickness. For 10 MHz ultrasonic insonation of a polymethylmethacrylate (PMMA) tube with an inner diameter of 195 microm and an outer diameter of 260 microm, the peak pressure within the tube can be up to 300% of incident pressure amplitude. However, using 1 MHz ultrasound and a silica tube with an inner diameter of 12 microm and an outer diameter of 50 microm, the peak pressure within the tube is only 12% of the incident pressure amplitude and correspondingly, the spatial-average-time-average intensity within the tube is only 1% of the incident intensity. PMID- 18395963 TI - Chronic pelvic pain and lower urinary tract symptoms in both sexes: analysis of 2749 participants of an urban health screening project. AB - BACKGROUND: Recent studies question the role of the prostate as the key factor in the pathogenesis of chronic pelvic pain syndrome (CPPS). OBJECTIVE: To compare symptoms related to CPPS and lower urinary tract symptoms (LUTS) in both sexes. DESIGN, SETTING, AND PARTICIPANTS: Participants of a voluntary health examination in Vienna. INTERVENTION AND MEASUREMENTS: All participants completed a detailed questionnaire containing the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI), with the female homolog of each male anatomical term use on questionnaires for female participants, the International Prostate Symptom Score (IPSS), and additional questions on pelvic pain. Furthermore, all participants underwent a detailed health investigation performed by a general practitioner. RESULTS AND LIMITATIONS: The study cohort comprised 1768 men and 981 women. The mean NIH-CPSI was 7.2+/-0.1 in women and 3.8+/-0.2 in men (p<0.001). In subject up to the age of 70 yr, the NIH-CPSI was higher in women (p<0.001). The NIH-CPSI increased with age in men (p<0.001), yet not in women (p=0.4). The prevalence of symptoms suggestive of CPPS in this selected population was 5.7% in women and 2.7% in men, and was higher in premenopausal women (p=0.03). Until the age of 50 yr, NIH-CPSI pain score in women exceeded that of men (p<0.001). The mean IPSS was higher in women (p<0.001). Storage symptoms were higher in women up to 60 yr, and voiding symptoms were higher in men above 60 yr. In men and women with symptoms suggestive of CPPS, the mean IPSS was significantly higher compared with those without CPPS symptoms (p<0.001). Limitations of our study are (1) that a urological evaluation was not performed and (2) that the questionnaire was not formerly validated for females. CONCLUSION: The preponderance of CPPS in females raises questions about the etiological role of the prostate in all cases with chronic pelvic pain and suggests that other pathomechanisms are likely to be involved. PMID- 18395964 TI - Generation of a functional, differentiated porcine urothelial tissue in vitro. AB - BACKGROUND: The primary function of urothelium is to serve as a physical urinary barrier. This function is dependent on features expressed at the molecular level that are acquired during cytodifferentiation. Urothelial cells lose differentiated and functional characteristics when propagated in vitro. OBJECTIVE: To investigate methods of inducing molecular and functional differentiation of normal porcine urothelial (NPU) cells in vitro. DESIGN AND MEASUREMENTS: NPU cells were isolated from normal porcine bladders and propagated in a low-calcium keratinocyte serum-free medium. Effects of 5% fetal bovine serum (FBS) and exogenous calcium were investigated. Molecular differentiation was assessed by immunolabelling for urothelial differentiation-associated proteins (UPIIIa, CK20, ZO-1), and barrier function was assessed by measurement of transepithelial electrical resistance (TER). RESULTS: NPU cell cultures grew as monolayers in low-calcium, serum-free medium. Supplementation with 5% FBS and/or physiological calcium resulted in stratification into basal, intermediate, and superficial cell zones. Superficial cells were positive for UPIIIa, CK20, and ZO 1. TER measurement showed that NPU cells grown with FBS had significantly enhanced barrier function (6,720 ohms.cm(2)+/-1312 SD) compared with cells grown without FBS (102 ohms.cm(2)+/-34 SD; p<0.001). LIMITATIONS: Importantly, our study demonstrates that expression of differentiation-associated immunohistochemical markers by cultured urothelial cells can be regarded as evidence of only morphological differentiation and does not represent a surrogate marker of function. CONCLUSIONS: We have shown that normal porcine bladder urothelium has many cell biological properties equivalent to normal human urothelium, making it an excellent research substitute for difficult-to-obtain tissue. A differentiated, functional barrier urothelium has been produced from porcine bladder urothelial cells propagated in vitro and displays molecular and functional properties equivalent to native urothelium. This tissue has application in developing tissue-engineered bladders with urinary barrier properties and as a research tool for understanding the relationship between molecular and functional tissue differentiation. PMID- 18395965 TI - Editorial comment on: Chronic pelvic pain and lower urinary tract symptoms in both sexes: analysis of 2749 participants of an urban health screening project. PMID- 18395966 TI - Editorial comment on: Chronic pelvic pain and lower urinary tract symptoms in both sexes: analysis of 2749 participants of an urban health screening project. PMID- 18395967 TI - Identification of SPAM messages using an approach inspired on the immune system. AB - In this paper, an immune-inspired model, named innate and adaptive artificial immune system (IA-AIS) is proposed and applied to the problem of identification of unsolicited bulk e-mail messages (SPAM). It integrates entities analogous to macrophages, B and T lymphocytes, modeling both the innate and the adaptive immune systems. An implementation of the algorithm was capable of identifying more than 99% of legitimate or SPAM messages in particular parameter configurations. It was compared to an optimized version of the naive Bayes classifier, which has been attained extremely high correct classification rates. It has been concluded that IA-AIS has a greater ability to identify SPAM messages, although the identification of legitimate messages is not as high as that of the implemented naive Bayes classifier. PMID- 18395968 TI - A PPAR-independent pathway to PUFA-induced COX-2 expression. AB - Polyunsaturated fatty acids (PUFAs) induce COX-2 in bovine endometrial stromal cells through activation of peroxisome-proliferator-activated receptor alpha (PPARalpha). We have investigated alternative (PPAR-independent) pathways to COX 2 induction using a reporter construct driven by a COX-2 gene promoter sequence lacking a PPAR response element. This construct was induced by PUFAs, but not by PPAR agonists. PPAR-independent reporter gene expression occurred 6h after PPAR dependent induction of the endogenous COX-2 gene. In contrast to PPAR-dependent COX-2 induction, which is not affected by NF-kappaB inhibitors, the PPAR independent pathway was blocked by the NF-kappaB inhibitor MG132 or following deletion of NF-kappaB sites in the COX-2 promoter. The PPAR-independent effect of PUFA was mimicked by the PKC activators 4beta-PMA and prostaglandin F(2alpha), but was not blocked by the PKC inhibitor RO318425. The results demonstrate a pathway to the induction of COX-2 by PUFAs requiring NF-kappaB but not PPAR or PKC. PMID- 18395969 TI - Somatostatin: the beginnings, 1972. PMID- 18395970 TI - Signal-dependent regulation of gene expression as a target for cancer treatment: inhibiting p38alpha in colorectal tumors. AB - In the last year, several evidences indicated that pharmacological manipulation of relevant signaling pathways could selectively affect gene expression to influence cell fate. These findings render of extreme importance the elucidation of how external stimuli are transduced to mediate chromatin modifications, resulting in a permissive or repressive environment for gene expression. These signaling cascades activate or repress the function of chromatin binding proteins that represent attractive pharmacological targets for human diseases. Actually, the closer the target is to chromatin, the more the transcriptional effect will be selective. Recent studies suggest that pharmacological manipulation of signaling pathways to modulate cell fate is indeed possible and that chromatin associated kinases could represent an optimal target. The p38 MAPK is the prototype of this class of enzymes and its central role in the transcription process is evolutionary conserved. In this review we will focus on the possibility to inhibit p38alpha in colorectal cancer to arrest tumor progression and induce autophagic cell death. PMID- 18395972 TI - A novel function of CD82/KAI-1 on E-cadherin-mediated homophilic cellular adhesion of cancer cells. AB - In this study, we analyzed the effect of the metastasis suppressor CD82/KAI-1, a member of the tetraspanin superfamily, on intercellular adhesion on cancer cells. The newly established invasion assay and the cell aggregation assay revealed that CD82 strengthens E-cadherin-mediated intercellular adhesion. Interestingly, ectopic expression of CD82 stabilized E-cadherin/beta-catenin complex formation. Furthermore, CD82 reduced tyrosine phosphorylation of beta-catenin on HGF stimulation. Taken together, CD82 may stabilize or strengthen E-cadherin dependent intercellular adhesion by regulating beta-catenin-mediated signal transduction on cancer cells, and consequently, prevent cancer cells from seceding from the primary tumor site. PMID- 18395971 TI - c-Met gene amplification is associated with advanced stage colorectal cancer and liver metastases. AB - The c-Met proto-oncogene encodes a receptor tyrosine kinase (TK) that promotes invasive tumor growth and metastasis. Recent studies show that the presence of c Met gene amplification is predictive for selective c-Met TK inhibitors in gastric cancer and lung cancer. In this study, we utilized a highly quantitative PCR/ligase detection reaction technique to quantify c-Met gene copy number in primary colorectal cancer (CRC) (N=247), liver metastases (N=147), and paired normal tissues. We identified no differences in c-Met gene copy number between normal colonic mucosa and liver tissue. However, mean c-Met gene copy number was significantly elevated in CRC compared with normal mucosa (P<0.001), and in liver metastases compared with normal liver (P<0.001). Furthermore, a significant increase in c-Met was seen in liver metastases compared with primary CRC (P<0.0001). c-Met gene amplification was observed in 2% (3/177) of localized cancers, 9% (6/70) of cancers with distant metastases (P<0.02), and 18% (25/147) of liver metastases (P<0.01). Among patients treated by liver resection, there was a trend toward poorer 3-year survival in association with c-Met gene amplification (P=0.07). Slight increases in c-Met copy number can be detected in localized CRCs, but gene amplification is largely restricted to Stage IV primary cancers and liver metastases. c-Met gene amplification is linked to metastatic progression, and is a viable target for a significant subset of advanced CRC. PMID- 18395973 TI - Perifosine induces differentiation and cell death in prostate cancer cells. AB - We analyzed the mechanism of action for perifosine (D-21266), a new synthetic alkylphospholipid Akt inhibitor, using LNCaP and PC-3 prostate cancer cells. Perifosine treatment of PC-3 cells resulted in cytostatic and cytotoxic effects. Cytostatic effects were characterized by cell growth arrest, cell cycle block, and morphological changes, such as a cell enlargement and granulation, hallmarks of differentiating PC-3 cells. Specific differentiation markers including prostasomal, secretory and plasma membrane proteins, and keratins were induced by perifosine. Among them, we detected strong induction and secretion of CEACAM5 protein. In contrast, perifosine strongly reduced caveolin-1 RNA levels. Cytotoxic effects included para-apoptosis, apoptosis, and necrosis. To pursue the mechanisms responsible for these activities we focused on signaling pathways that lie downstream of Akt. Perifosine-triggered GSK-3beta activation in PC-3 and LNCaP cells resulted in the expression of GSK-3beta-related differentiation markers. This expression was reduced in the presence of specific siRNA for GSK 3beta or for its target CREB protein. The use of the GSK-3beta inhibitor lithium chloride indicated that GSK-3beta partially protects prostate cancer cells from the cytotoxic effects of perifosine. Together, these findings indicate that perifosine induces GSK-3beta-related differentiation and caspase-independent cell death in prostate cancer PC-3 cells. In addition our results identify specific biomarkers for perifosine therapy. PMID- 18395974 TI - Quantitative and qualitative characterization of plasma DNA identifies primary and recurrent colorectal cancer. AB - Because plasma DNA may be a useful tool for cancer detection, we screened primary tumors and related multiple plasma samples at the time of surgery and during the follow-up period for plasma DNA level as well as for K-Ras mutations and p16INK4a promoter hypermethylation in colorectal cancer patients. At the time of surgery, DNA levels were higher in tumor patients than in healthy donors, and K-Ras and p16INK4a alterations were detected in 7 and 11 cancers respectively, and in all related plasma samples. During the follow-up, plasma DNA levels decrease progressively but rapidly increased when a relapse occurred, whereas K-Ras and p16INK4a alterations were detected only in relapsed patients. Therefore, combined quantitative and qualitative analyses of plasma DNA confirm the presence of colorectal cancer, define disease-free status and indicate the presence of relapse. PMID- 18395975 TI - Inherent safety key performance indicators for hydrogen storage systems. AB - The expected inherent safety performance of hydrogen storage technologies was investigated. Reference schemes were defined for alternative processes proposed for hydrogen storage, and several storage potentialities were considered. The expected safety performance of alternative process technologies was explored estimating key performance indicators based on consequence assessment and credit factors of possible loss of containment events. The results indicated that the potential hazard is always lower for the innovative technologies proposed for hydrogen storage, as metal or complex hydrides. This derived mainly from the application of the inherent safety principles of "substitution" and "moderation", since in these processes hydrogen is stored as a less hazardous hydride. However, the results also evidenced that in the perspective of an industrial implementation of these technologies, the reliability of the auxiliary equipment will be a critical issue to be addressed. PMID- 18395976 TI - Synthesized layered inorganic-organic magnesium organosilicate containing a disulfide moiety as a promising sorbent for cations removal. AB - A new-layered inorganic-organic magnesium organosilicate was synthesized through a single step template sol-gel route under mild conditions, using a new alkoxysilane, containing a 2-aminophenyldisulfide molecule. Elemental analysis data based on the nitrogen atom showed an incorporation of 1.97mmol of organic pendant groups for each gram of the hybrid formed. The X-ray diffraction patterns demonstrated that this nanocompound exhibited lamellar structure, in agreement with that found for natural inorganic silicates. Infrared spectroscopy and nuclear magnetic resonance for the (29)Si nucleus in the solid state are in agreement with the success of the proposed synthetic method. The presence of nitrogen and sulfur basic centers attached to the pendant groups inside the lamellar structure is used as basic centers to coordinate cations from aqueous solution at the solid/liquid interface. The isotherms were fitted to Langmuir and Freundlich models. The maxima adsorption capacities for copper, lead and cadmium, calculated from Langmuir model, were 3.28, 1.42 and 0.35mmol g(-1), respectively. These values are comparable to other adsorbing nanomaterials. This behavior suggested that this new inorganic-organic hybrid could be employed as a promising adsorbent for cation removal from polluted systems. PMID- 18395977 TI - Photolytic decolorization of Rose Bengal by UV/H(2)O(2) and data optimization using response surface method. AB - Rose Bengal (C.I. name is Acid Red 94) was irradiated with UV light in the presence of hydrogen peroxide. The photoinduced decolorization of the dye was monitored spectrophotometrically. The apparent rate of decolorization was calculated from the observed absorption data and was found to be pseudo first order. A systematic study of the effect of dye concentration and H(2)O(2) concentration on the kinetics of dye decolorization was also carried out. Dye decolorization increased with increasing H(2)O(2) concentration and decreasing dye concentration. The maximum dye decolorization was determined as 90% with 0.005 mM dye at optimum 0.042 M H(2)O(2) and pH 6.6. Additionally, the effect on decolorization of this dye in the presence of some additives (ions) was also investigated. It was seen that sulphite caused a maximum effect on % decolorization of the dye solution. A plausible explanation involving the probable radical initiated mechanism was given to explain the dye decolorization. The experimental data was also optimized using the response surface methodology (RSM). According to ANOVA results, the proposed model can be used to navigate the design space. It was found that the response of Rose Bengal degradation is very sensitive to the independent factors of dye concentration, H(2)O(2) concentration, pH and reaction time. The proposed model for D-optimal design fitted very well with the experimental data with R(2) and R(adj)(2) correlation coefficients of 0.85 and 0.80, respectively. PMID- 18395978 TI - Expression profile of PRAF2 in the human brain and enrichment in synaptic vesicles. AB - PRA1 domain family, member 2 (PRAF2) is a novel 19-kDa protein with a prenylated Rab acceptor 1 (PRA1) motif and four transmembrane domains. Our previous studies revealed that PRAF2 is highly expressed in the brain and serves as a candidate prognostic marker in neuroblastoma (NB). PRAF2 is related to proteins PRAF1 (PRA1, prenylin, Yip3) and PRAF3 (GTRAP3-18, JWA, Arl6-IP5), both of which are enriched in the brain and implicated in cellular transport and endo/exocytic vesicle trafficking. However, the function for PRAF2 remains unknown. In this study, we analyzed the distribution and localization of PRAF2 in the mature human brain using two new antibodies specific for the protein. Analysis by immunohistochemistry revealed that in the human cerebellum, the PRAF2 protein was strongly expressed in Purkinje cells and, more moderately, in cells of the molecular and the granular layers. In the cerebral cortex, hippocampus, and lateral ventricles, PRAF2 protein was detected in neuronal cells, but not in non neuronal cells. Intriguingly, immunoblot analysis revealed that PRAF2 is enriched in synaptic vesicles (SVs) prepared from rat brains. The expression of PRAF2 in specific regions of the brain including SVs suggest an important physiological function for this novel protein, possibly by participating in multiple aspects of SV maturation, transport, and signal transmission. PMID- 18395979 TI - Unmyelinated tactile afferents have opposite effects on insular and somatosensory cortical processing. AB - A previous functional magnetic resonance imaging (fMRI) study of an A-beta deafferented subject (GL) showed that stimulation of tactile C afferents (CT) activates insular cortex whereas no activation was seen in somatosensory cortices. Psychophysical studies suggested that CT afferents contribute to affective but not to discriminative aspects of tactile stimulation. We have now examined cortical processing following CT stimulation in a second similarly deafferented subject (IW), as well as revisited the data from GL. The results in IW showed similar activation of posterior insular cortex following CT stimulation as in GL and so strengthen the view that CT afferents underpin emotional aspects of touch. In addition, CT stimulation evoked significant fMRI deactivation in somatosensory cortex in both subjects supporting the notion that CT is not a system for discriminative touch. PMID- 18395980 TI - Association between AKT1 gene and Parkinson's disease: a protective haplotype. AB - Variation in AKT1 has been associated with schizophrenia, bipolar disease and type II diabetes. The aim of the present study was to investigate the potential role of variability within AKT1 as a risk factor for Parkinson's disease (PD). We performed a case-control association analysis of AKT1 in a Greek cohort of PD using four tagging SNPs and five constructed haplotypes. To assess the structure of this locus in different populations we have performed linkage disequilibrium (LD) analysis using these variants [dunning]. In multilocus analysis, the frequency of a four-SNP1/2/3/4 haplotype was significantly higher in controls in comparison with PD patients (chi(2)=19.76, p=0.00009, OR=0.11 C.I.=0.03-0.35). The association remained significant even after Bonferroni correction for the number of haplotypes (p=0.0002). So, this certain haplotype was significantly associated with reduced risk of the disease. The data presented here suggest the involvement of AKT1 in protection of PD through many possible mechanisms involving different signaling pathways that could be potential therapeutic targets in the future. PMID- 18395981 TI - Overexpression of APP stimulates basal and constitutive exocytosis in PC12 cells. AB - The mechanisms that underlie the altered neurotransmitter system in Alzheimer's disease (AD) are not well understood. Amyloid precursor protein (APP) is a precursor protein for beta-amyloid, an important trigger protein in the pathogenesis of AD. Duplication of the APP gene as well as APP genes that contain certain mutations has been reported to be associated with familial AD (FAD), and a role of APP in neurotransmission has been suggested recently. This study examines the role of APP in exocytosis in PC12 cells using transfected human growth hormone (hGH) as a reporter for secretion. It was found that overexpression of APP or expression of the Swedish FAD mutation (APPsw) in PC12 cells significantly increased the basal secretion and constitutive secretion of hGH. Expression of an APP phosphorylation-deficient mutant decreased both basal and constitutive secretion relative to the APP wild-type, suggesting a role for APP-Thr668 phosphorylation in secretion in PC12 cells. Overexpression of X11alpha, a protein that stabilizes cellular APP, also increased the basal secretion of hGH but, contrary to APP, decreased the constitutive secretion of hGH, suggesting that basal and constitutive secretion is likely to proceed via distinct pathways and that the increase in the basal secretion of hGH may result from APP-X11alpha interaction. These results demonstrate an unknown role for APP in secretion, and suggest that elevated levels of APP or APP mutation in FAD brains contribute to the altered neurotransmitter pathology of AD through stimulation of basal and constitutive secretion. PMID- 18395982 TI - Doctors' attitudes and beliefs regarding acute low back pain management: A systematic review. AB - The aim of this systematic review was to determine the attitudes and beliefs of doctors to acute low back pain, and the factors that influence these. The review comprised three phases: a methodological assessment of databases (Medline, EMBASE, Psychinfo, BIOSIS, CINAHL, and the Cochrane Central Register of Controlled Trials) identified potential papers; these were screened for inclusion criteria by two independent reviewers, the extraction of data and the rating of internal validity and strength of the evidence, using valid and reliable scales from accepted papers. Themes were then identified from the accepted literature. The search generated a total of 15 papers of both qualitative (n=3) and quantitative (n=12) methodologies. Themes that emerged included doctors' attitudes and beliefs, and four factors that influenced attitudes and beliefs: doctors' specialty, demographic factors, personal beliefs and education. There was consistent evidence that doctors' specialty impacted their attitudes and beliefs: lack of consensus regarding the natural history of LBP, around treatment options, and issues regarding work. There was inconsistent evidence that demographic factors (age) and level of education impacted doctors' attitudes and beliefs. Strategies to address/ modify these attitudes and beliefs are required, as in some cases they are at odds with guideline recommendations. Long term, these changes in these areas have the potential to maximise patient-care, and reduce costs to health services. PMID- 18395983 TI - C-fiber spontaneous discharge evoked by chronic inflammation is suppressed by a long-term infusion of lidocaine yielding nanogram per milliliter plasma levels. AB - Nociceptors innervating inflamed tissue acquire an abnormal spontaneous discharge that is believed to be at least part of the reason for the persistent spontaneous pain, allodynia, and hyperalgesia that accompany inflammation. Recent studies suggest that patients with chronic inflammatory pain may obtain an analgesic effect with transdermal application of lidocaine that yields very low plasma levels (130-225 ng/ml). The aim of this study was to investigate whether a 7-day exposure to such low plasma levels of lidocaine had an effect on inflammation evoked spontaneous discharge in the rat. Seven days after a hind-paw injection of Complete Freund's Adjuvant (CFA), we confirmed the presence of myositis, pain hypersensitivity, and a significant increase in the incidence of spontaneous discharge in A-fibers and C-fibers innervating both muscle and skin. We then compared the incidence of spontaneous discharge in muscle and cutaneous fibers in CFA-injected animals treated with a 7-day infusion of saline or lidocaine. The lidocaine infusion yielded a plasma level of 210 ng/ml. The muscle C-fiber discharge was completely inhibited by the lidocaine infusion and the cutaneous C fiber discharge was suppressed by 50%. Lidocaine infusion had no effect on the incidence of spontaneous discharge in muscle or cutaneous A-fibers. Lidocaine infusion reduced mechano-hyperalgesia but had no effect on mechano-allodynia or heat-hyperalgesia. We conclude that the analgesic effects seen clinically with transdermal lidocaine administration yielding low plasma levels may be due to a systemic drug action on spontaneously active C-fibers. PMID- 18395984 TI - Symmetry in electron diffractions from helical structures. AB - In this work, we elucidate the regular rule of the symmetry in electron diffraction patterns from helical structures. It is affected by the value of the dominating Bessel function orders for the layer lines and relative orientation of the samples with respect to the electron beam. For Single-Walled Carbon nanotubes (SWCNTs), we use analytic analysis and computer simulations to demonstrate that the 2mm symmetry of the electron diffraction may break down not only from an achiral SWCNT, but also from a chiral SWCNT. Also, the simulation work for B-DNA is presented to corroborate the theoretical analysis. PMID- 18395986 TI - MRI induced fourth-degree burn in an extremity, leading to amputation. PMID- 18395987 TI - Lightning injury: a review. AB - Lightning is an uncommon but potentially devastating cause of injury in patients presenting to burn centers. These injuries feature unusual symptoms, high mortality, and significant long-term morbidity. This paper will review the epidemiology, physics, clinical presentation, management principles, and prevention of lightning injuries. PMID- 18395988 TI - Pharmacokinetic variability of extended interval tobramycin in burn patients. AB - BACKGROUND: Aminoglycosides are mandatory in the treatment of severe infections in burns. However, their pharmacokinetics are difficult to predict in critically ill patients. Our objective was to describe the pharmacokinetic parameters of high doses of tobramycin administered at extended intervals in severely burned patients. METHODS: We prospectively enrolled 23 burned patients receiving tobramycin in combination therapy for Pseudomonas species infections in a burn ICU over 2 years in a therapeutic drug monitoring program. Trough and post peak tobramycin levels were measured to adjust drug dosage. Pharmacokinetic parameters were derived from two points first order kinetics. RESULTS: Tobramycin peak concentration was 7.4 (3.1-19.6)microg/ml and Cmax/MIC ratio 14.8 (2.8-39.2). Half-life was 6.9 (range 1.8-24.6)h with a distribution volume of 0.4 (0.2 1.0)l/kg. Clearance was 35 (14-121)ml/min and was weakly but significantly correlated with creatinine clearance. CONCLUSION: Tobramycin had a normal clearance, but an increased volume of distribution and a prolonged half-life in burned patients. However, the pharmacokinetic parameters of tobramycin are highly variable in burned patients. These data support extended interval administration and strongly suggest that aminoglycosides should only be used within a structured pharmacokinetic monitoring program. PMID- 18395989 TI - A non-surgical device for faecal diversion in the management of perineal burns. AB - BACKGROUND: Burns to the perineal, buttock and upper thigh areas are frequently exposed to continual faecal contamination which results in sepsis, graft loss, delayed wound healing and shrinkage of scars. A temporary diverting colostomy may be required. Two specifically designed intrarectal catheters were evaluated for their safety and ability to divert faeces away from the burn and allow wound healing. METHODS: A prospective study was conducted involving patients at the burns centre. Either the Zassi Bowel Management System or the Flexi-Seal Fecal Management System were used. These differed only in the presence of a specific intraluminal balloon in the Zassi system to facilitate retention of infused irrigates. Data regarding skin graft success, wound contamination and adverse events were collected. RESULTS: The study included eight participants, five of whom were treated successfully without colostomy. Four participants experienced complications, comprising one bowel occlusion, one anal ulceration and two reversible cases of anal atony. CONCLUSION: A specifically designed intrarectal catheter can divert faeces to allow wound healing, and may avert colostomy. More studies are necessary to evaluate safety. PMID- 18395990 TI - Cultured keratinocytes in fibrin with decellularised dermis close porcine full thickness wounds in a single step. AB - The purpose of this study was to assess the feasibility of combined keratinocyte and dermal scaffold transplantation performed in a single step for treatment of full-thickness wounds. Cultured autologous keratinocytes were suspended in fibrin and grafted together with decellularised human dermis (Alloderm) in a porcine animal model, involving 10 animals over 4 weeks. Wound healing was evaluated by planimetry. Histology included morphological analysis as well as immunohistochemistry at regular intervals (1, 2 and 4 weeks). The results showed both successful histo-integration of the in vivo composite grafts and reduced wound contraction, compared with the control group (plain epithelial grafts). Histologically a neo-epithelium originated from the grafted cells on top of the decellularised dermis, as well as a reconstituted basement membrane. After 4 weeks cellular ingrowth into the dermal matrix could be observed. The successful combination of a keratinocyte-fibrin suspension and acellular dermis applied in a single step onto full-thickness wounds resulted in closure. PMID- 18395991 TI - Comparison of premortem clinical diagnosis and autopsy findings in patients with burns. AB - INTRODUCTION: Despite the diagnostic advancements, some clinically important diagnoses remain undetected during intensive care in burn patients. The aim of this study was to compare the premortem clinical diagnoses and autopsy findings. PATIENTS AND METHODS: A retrospective review of all burn deaths during 1995-2005 was conducted. The clinical diagnoses and autopsy reports were reviewed, and diagnostic discrepancies were classified into four categories, according to the impact on the treatment. RESULTS: Overall mortality during the study period was 5.4%. Altogether 74 deaths were recorded, of which 71 were included in the study. Typical patient was a 58-year-old male with flame burn of %TBSA 49, ABSI 10. Clinical diagnostic discrepancies were found in 14.1% of the patients; one diagnostic discrepancy was recorded in each of the patients. Of these diagnostic discrepancies, 8.5% were considered major, and 5.6% would have altered the clinical outcome or therapy, if known at the time. Diagnostic discrepancies consisted of one cardiovascular, seven respiratory and two gastrointestinal missed diagnosis. The most common missed diagnosis was pneumonia. CONCLUSION: This study emphasizes the usefulness of autopsies to provide valuable clinical data for the treatment of burn patients. It also highlights the few missed diagnoses which may occur in burn patients. PMID- 18395992 TI - Identification of capsaicin-sensitive rectal mechanoreceptors activated by rectal distension in mice. AB - Rodents detect visceral pain in response to noxious levels of rectal distension. However, the mechanoreceptors that innervate the rectum and respond to noxious levels of rectal distension have not been identified. Here, we have identified the mechanoreceptors of capsaicin-sensitive rectal afferents and characterized their properties in response to circumferential stretch of the rectal wall. We have also used the lethal spotted (ls/ls) mouse to determine whether rectal mechanoreceptors that respond to capsaicin and stretch may also develop in an aganglionic rectum that is congenitally devoid of enteric ganglia. In wild type (C57BL/6) mice, graded increases in circumferential stretch applied to isolated rectal segments activated a graded increase in firing of slowly-adapting rectal mechanoreceptors. Identical stimuli applied to the aganglionic rectum of ls/ls mice also activated similar graded increases in firing of stretch-sensitive rectal afferents. In both wild type and aganglionic rectal preparations, focal compression of the serosal surface using von Frey hairs identified mechanosensitive "hot spots," that were associated with brief bursts of action potentials. Spritzing capsaicin (10 microM) selectively onto each identified mechanosensitive hot spot activated an all or none discharge of action potentials in 32 of 56 identified hot spots in wild type mice and 24 of 62 mechanosensitive hot spots in the aganglionic rectum of ls/ls mice. Each single unit activated by both capsaicin and circumferential stretch responded to low mechanical thresholds (1-2 g stretch). No high threshold rectal afferents were ever recorded in response to circumferential stretch. Anterograde labeling from recorded rectal afferents revealed two populations of capsaicin-sensitive mechanoreceptor that responded to stretch: one population terminated within myenteric ganglia, the other within the circular and longitudinal smooth muscle layers. In the aganglionic rectum of ls/ls mice, only the i.m. mechanoreceptors were identified. Both myenteric and i.m. mechanoreceptors could be identified by their immunoreactivity to the anti-TRPV1 antibody and the vesicular glutamate transporter, Vglut2. Myenteric mechanoreceptors had a unique morphology, consisting of smooth bulbous nodules that ramified within myenteric ganglia. In summary, the rectum of wild type mice is innervated by at least two populations of capsaicin-sensitive rectal mechanoreceptor, both of which respond to low mechanical thresholds within the innocuous range. These findings suggest that the visceral pain pathway activated by rectal distension is likely to involve low threshold rectal mechanoreceptors that are activated within the normal physiological range. PMID- 18395993 TI - Anandamide and neutrophil function in patients with fibromyalgia. AB - Fibromyalgia (FM) is a common stress-related painful disorder. There is considerable evidence of neuroimmunologic alterations in FM which may be the consequence of chronic stress and pain or causally involved in the development of this disorder. The endocannabinoid system has been shown to play a pivotal role in mammalian nociception, is activated under stressful conditions and can be an important signaling pathway for immune modulation. The endocannabinoid system could therefore be involved in the complex pathophysiology of FM. We tested this hypothesis by evaluating the effects of stress hormones and the endocannabinoid anandamide on neutrophil function in patients with FM. We determined plasma levels of catecholamines, cortisol and anandamide in 22 patients with primary FM and 22 age- and sex-matched healthy controls. Neutrophil function was characterized by measuring the hydrogen peroxide (H2O2) release (oxidative stress) and the ingestion capabilities of neutrophils (microbicidal function). FM patients had significantly higher norepinephrine and anandamide plasma levels. Neutrophils of FM patients showed an elevated spontaneous H2O2 production. The ability of neutrophils to adhere was negatively correlated with serum cortisol levels. Adhesion and phagocytosis capabilities of neutrophils correlated positively with anandamide plasma levels. In conclusion, patients with FM might benefit from pharmacologic manipulation of endocannabinoid signaling which should be tested in controlled studies. PMID- 18395994 TI - [Venous thromboembolism associated with long-term use of central venous catheters in cancer patients]. AB - Increased incidence of cancers and the development of totally implanted venous access devices that contain their own port to deliver chemotherapy will lead to a greater than before numbers of central venous catheter-related thrombosis (CVCT). Medical consequences include catheter dysfunction and pulmonary embolism. Vessel injury caused by the procedure of CVC insertion is the most important risk factor for development of CVCT. This event could cause the formation of a fresh thrombus, which is reversible in the large majority of patients. In some cases, thrombus formation is not related to catheter insertion. The incidence of CVC related DVT assessed by venography has been reported to vary from 30 to 60% but catheter-related DVT in adult patients is symptomatic in only 5% of cases. The majority of patients with CVC-related DVT is asymptomatic or has nonspecific symptoms: arm or neck swelling or pain, distal paresthesias, headache, congestion of subcutaneous collateral veins. In the case of clinical suspicion of CVC related deep venous thrombosis (DVT), compressive ultrasonography (US), especially with doppler and color imaging, currently is first used to confirm the diagnosis. Consequently, contrast venography is reserved for clinical trials and difficult diagnostic situations. There is no consensus on the optimal management of patients with CVC-related DVT. Treatment of CVC-related VTE requires a five- to seven-day course of adjusted-dose unfractionated heparin or low molecular weight heparin (LMWH) followed by oral anticoagulants. Long-term LMWH that has been shown to be more effective than oral anticoagulant in cancer patients with lower limb DVT, could be used in these patients. The efficacy and safety of pharmacologic prophylaxis for CVC related thrombosis is not established and the last recommendations suggest that clinicians not routinely use prophylaxis to try to prevent thrombosis related to long-term indwelling CVCs in cancer patients. Additional studies performed in high risk populations with appropriate dosage and timing will help to define which patients could benefit from prophylaxis. PMID- 18395995 TI - A simultaneous procedure facilitates acquisition under an optimal interstimulus interval in artificial neural networks and rats. AB - In a computer simulation, a neural network first received a simultaneous procedure, where the interstimulus interval (ISI) was 0 time-steps (ts). Output activations were near zero under this procedure. The network then received a forward-delay procedure where the ISI was 8 ts. Output activations increased to the near-maximum level faster than those of a control network that first received an explicitly unpaired procedure. Comparable results were obtained with rats that first received trials where a retractable lever was presented for 3s concurrently with access to water. Low-lever pressing was observed under this procedure. The rats then received trials where the lever was followed 15s after by water. Lever pressing appeared faster than a control group that received the 15-s ISI after an explicitly unpaired procedure. The model used in the simulation explains these results as connection-weight increments that promote little output activations in a simultaneous procedure, but facilitate acquisition in an optimal ISI. PMID- 18395996 TI - Caffeine induction of Cyp6a2 and Cyp6a8 genes of Drosophila melanogaster is modulated by cAMP and D-JUN protein levels. AB - Cytochrome P450 monooxygenases or CYPs, a family of endobiotics and xenobiotics metabolizing enzymes, are found in all organisms. We reported earlier that the promoters of Drosophila Cyp6a2 and Cyp6a8 genes are induced by caffeine. Since caffeine antagonizes adenosine receptor (AdoR) and inhibits cAMP phosphodiesterase (PDE), we used luciferase reporter gene to examine whether in SL-2 cells and adult Drosophila, induction of the two Cyp6 genes is mediated via AdoR and/or PDE pathway. Results showed that AdoR is not involved because AdoR agonists or antagonists do not affect the Cyp6 promoter activities. However, inhibition of PDE by specific inhibitors including caffeine causes induction of both Cyp6 gene promoters. We also found that flies mutant for dunce gene coding for cAMP-PDE, have higher Cyp6a8 promoter activity than the wild-type flies. We demonstrate that caffeine treatment increases intracellular cAMP levels, and cAMP treatment induces the Cyp6 gene promoters. Since both Cyp6 genes have multiple sites for JUN transcription factors, which generally play a positive role in cAMP pathway, effect of Drosophila jun (D-jun) on the Cyp6a8 promoter activity was examined. Results showed that the expression of D-jun sense plasmid causes downregulation rather than activation of the Cyp6a8 promoter. Conversely, expression of antisense plasmid increased the promoter activity. Interestingly, caffeine treatment decreased the D-JUN protein level in SL-2 cells as well as in adult flies. These results suggest that D-jun acts as a negative regulator, and caffeine induction of Cyp6a8 and Cyp6a2 genes is mediated by the upregulation of cAMP pathway and downregulation of the D-JUN protein level. PMID- 18395997 TI - Expression analysis of the calcineurin B-like gene family in rice (Oryza sativa L.) under environmental stresses. AB - Calcium plays a crucial role as a second messenger in mediating various defense responses under environmental stresses. Calcineurin B-like (CBL) proteins have been implicated as important Ca2+ sensors in plant-specific calcium signaling. Based on the similarity of sequence, ten CBL genes were identified by searching the rice japonica genome database, which were randomly distribute on chromosomes 1, 2, 3, 5, 10 and 12. By semi-quantitative RT-PCR approach the expression pattern of each gene was detected in various organs at the adult stage and seedlings treated with NaCl, PEG and cold stresses or exogenous ABA. The results showed that the induction of each rice CBL gene was not only responsive to different stress conditions, but also organ specific. In vivo targeting experiment revealed that OsCBL8 localized to the plasma membrane, which was consistent with OsCBL4 and SOS3 previously reported. To elucidate the putative function of OsCBL8 gene, transgenic rice plants overexpressing OsCBL8 gene were generated by the Agrobacterium-mediated approach. The OsCBL8 transgenic rice seedlings showed more tolerance to salt stress than non-transgenic seedlings. PMID- 18395998 TI - Genetic heterogeneity of bovine viral diarrhoea virus (BVDV) isolates from Turkey: identification of a new subgroup in BVDV-1. AB - Genetic heterogeneity of Turkish ruminant pestiviruses was investigated by phylogenetic analysis of complete N(pro) encoding nucleotide sequences. A total of 30 virus isolates obtained from 15 provinces around the country between 1997 and 2005 were included in the phylogenetic analysis. Virus isolates mostly originated from cattle with one isolate from sheep. The bovine isolates all belonged to BVDV-1, the sheep isolate to BVDV-2. Fifteen isolates formed a new subgroup within BVDV-1, tentatively named BVDV-1l. The remaining bovine isolates were typed as BVDV-1a (n=4), BVDV-1b (n=4), BVDV-1d (n=3), BVDV-1f (n=2) and BVDV 1h (n=1). The isolates allocated to BVDV-1l originated from various geographical regions in different years. There was no correlation between genetic grouping and locations where isolates were obtained. Viruses originating from one farm in most cases belonged to the same subgroup (n=5). This study indicates that the newly detected subgroup BVDV-1l is predominant and widespread in Turkey. Moreover, an ovine virus isolate was identified as the first member of BVDV-2 reported in Turkey. A serological survey using samples from western Turkey indicated that BVDV-2 is also present in cattle. PMID- 18395999 TI - Formation of DHP-derived DNA adducts from metabolic activation of the prototype heliotridine-type pyrrolizidine alkaloid, heliotrine. AB - Pyrrolizidine alkaloid-containing plants are widespread in the world and may be the most common poisonous plants affecting livestock, wildlife, and humans. Pyrrolizidine alkaloids require metabolism to exert their genotoxicity and tumorigenicity. Our mechanistic studies have determined that metabolism of the retronecine-type (riddelliine, retrorsine, and monocrotaline), heliotridine-type (lasiocarpine), and otonecine-type (clivorine) tumorigenic pyrrolizidine alkaloids in vivo and/or in vitro all generates a common set of 6,7-dihydro-7 hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP)-derived DNA adducts responsible for tumor induction. All the pyrrolizidine alkaloids studied previously are diesters with an ester linkage at the C7 and C9 positions of the necine base. In this study, we report that F344 rat liver microsomal metabolism of heliotrine, a tumorigenic monoester bearing a hydroxyl group at the C7 of the necine base, resulted in the formation of the dehydroheliotridine (DHH) metabolite. When incubations of heliotrine were carried out in the presence of calf thymus DNA, the same set of DHP-derived DNA adducts was formed. These results support that DHP-derived DNA adducts are potential common biomarkers of pyrrolizidine alkaloid exposure and tumorigenicity. For comparison, the dehydroretronecine (DHR)-derived DNA adducts formed from metabolism of riddleiine, retrorsine, monocrotaline, riddelleiine N-oxide, and retrorsine N-oxide were measured in parallel; the levels of DHP-derived DNA adduct formation were in the order: riddelliine approximately retrorsine>monocrotaline>retrorsine N-oxide>or=riddelliine N oxide>heliotrine. PMID- 18396000 TI - The in vivo effects of Tulbhagia violacea on blood pressure in a salt-sensitive rat model. AB - AIM OF THE STUDY: The in vivo effects of Tulbhagia violacea on systemic arterial blood pressure and on the renin-angiotensin system in a Dahl salt-sensitive rat model were investigated. MATERIALS AND METHODS: Animals were treated for 14 days intraperitoneally as follows: Tulbhagia violacea (Tvl) (50mg/kg b.w.), captopril (Cap) (10mg/kg b.w.) or DMSO (Con). Baseline blood pressures were recorded prior to the commencement of the study and biweekly during the experimental period. Urine volume and sodium concentration were measured during the experimental period. On day 15, animals were anaesthetized (sodium thiopentane, 50mg/kg, i.p.), blood samples for aldosterone levels were taken and the kidneys removed for determining AT1a mRNA expression. RESULTS: Cap and Tvl groups showed significantly reduced AT1a mRNA expressions by 3.11- and 5.03-fold, respectively, when compared to the Con group (p<0.05). When compared to baseline blood pressures (day 0); Cap and Tvl showed reductions in systolic blood pressure (SBP) of 7.76+/-0.41% and 9.12+/-0.31%, respectively (mean% decrease from day 0 to day 14). In contrast, in the Con group the systolic blood pressure increased from day 0 to day 14 by 4.66+/-0.56%. Blood pressure changes in all treated groups differed from Con significantly. Systolic blood pressure decreased with the decrease in AT1a mRNA expressions in these groups. When comparing day 0 to day 14, urine output increased in the Cap and Tvl groups. In the Con group, urinary volume was reduced by day 14 as compared to day 0. Urinary sodium excretion was increased in the treated groups by day 14. CONCLUSION: It can be concluded that Tulbhagia violacea reduces systemic arterial blood pressure in the Dahl rat by decreasing renal AT1 receptor gene expression and hence modulating sodium and water homeostasis. PMID- 18396001 TI - Recurrent facial nerve palsy associated with anti-GQ1b IgG antibodies. AB - Recurrent facial nerve palsy (RFNP) in childhood is usually considered to be a benign disorder. We report a 13-year-old female affected with RFNP in the absence of other neurologic signs, in which elevated serum IgG and IgM anti-GQ1b antibodies were detected. To our knowledge, this is the first example in the literature of RFNP, associated with anti-GQ1b IgG antibodies. The possible role of anti-GQ1b antibodies in isolated cranial neuropathy is discussed. PMID- 18396002 TI - A case of galactosialidosis with a homozygous Q49R point mutation. AB - Galactosialidosis is a rare lysosomal storage disease caused by a combined deficiency of lysosomal beta-galactosidase and neuraminidase, due to a primary defect in protective protein/cathepsin A. Three subtypes are recognized: the early infantile type, the late infantile type, and the juvenile/adult type. Here, we report a case of early infantile galactosialidosis in a female who was born at 31 weeks of gestation, after detection of fetal ascites at 21 weeks of gestation and development of fetal hydrops. After birth she received intensive treatment that led to improvement of edema and pleural effusion, but ascites slowly developed. She died of renal failure on day 207. An autopsy showed that all organs contained vacuolated cells, compatible with a storage disease. The patient had decreased activity of beta-galactosidase and undetectable neuraminidase activity in fibroblasts. A single A-G base transition at position 146 of exon 1 (Q49R) in protective protein/cathepsin A gene was found. The mutation has been reported previously in a Japanese patient with different phenotypes. However homozygous Q49R mutation detected in our case was severe prognosis. PMID- 18396003 TI - Idiopathic mesenteric venous thrombosis: report of a case. AB - INTRODUCTION: Idiopathic mesenteric venous thrombosis is a rare entity. An early diagnosis and thrombolytic and anticoagulant therapy are very important. PATIENT AND METHODS: We report a case of a patient, without any specific known risk factor, with small intestinal ischemia secondary to superior mesenteric vein thrombosis (SMVT). RESULTS: In our case, only a computed tomography (CT) abdominal scan permitted the diagnosis of SMVT. The patient was successfully treated by resection of the infarcted bowel with primary anastomosis and immediate postoperative anticoagulation. CONCLUSIONS: Diagnosis of intestinal ischemia from mesenteric venous thrombosis (MVT) is often delayed because the symptoms are nonspecific. Moreover, when there is not any known predisposing factor, the diagnosis may become even more difficult with significant morbidity and mortality. CT abdominal scan done early in case of nonspecific abdominal pain, since the patients had a previous history of venous thrombosis, may not require a surgical treatment of MVT. PMID- 18396004 TI - Volatilisation of metals and metalloids: an inherent feature of methanoarchaea? AB - As shown by recent studies, anaerobic members of Archaea and Bacteria are involved in processes that transform ionic species of metals and metalloids (arsenic, antimony, bismuth, selenium, tellurium and mercury) into volatile and mostly toxic derivatives (mainly methyl derivatives or hydrides). Since the fact that these transformations proceed in both environmental settings and in parts of the human body, we have to consider that these processes also interfere directly with human health. The diversity of the volatile derivatives produced and their emission rates were significantly higher in methanoarchaeal than in bacterial strains, which supports the pivotal role of methanoarchaea in transforming metals and metalloids (metal(loid)s) into their volatile derivatives. Compared with methanoarchaea, 14 anaerobic bacterial strains showed a significantly restricted spectrum of volatilised derivatives and mostly lower production rates of volatile bismuth and selenium derivatives. Since methanoarchaea isolated from the human gut (Methanosphaera stadtmanae, Methanobrevibacter smithii) showed a higher potential for metal(loid) derivatisation compared to bacterial gut isolates, we assume that methanoarchaea in the human gut are mainly responsible for the production of these volatile derivatives. The observation that trimethylbismuth ((CH(3))(3)Bi), the main volatile derivative of bismuth produced in human feces, inhibited growing cultures of Bacteroides thetaiotaomicron, a representative member of the human physiological gut flora, suggests that these volatiles exert their toxic effects on human health not only by direct interaction with host cells but also by disturbing the physiological gut microflora. PMID- 18396005 TI - RNAi-induced down-regulation of Mecp2 expression in the rat brain. AB - The MECP2 (methyl-CpG-binding protein 2) gene has been implicated in the pathogenesis of Rett Syndrome. A rat model with a down-regulated Mecp2 was established using a recombinant lentiviral vector expressing small hairpin RNA of the rat Mecp2 gene. Four recombinant vectors were constructed by inserting sequences of small hairpin RNA targeting the rat Mecp2 gene. To determine which vector resulted in optimal down-regulation, rat skin fibroblasts were transfected with four recombinant vectors, respectively. The recombinant vector mecp2-sh-1 decreased expression of Mecp2 mRNA and protein relative to the Control group. 12 24h after birth, neonatal rat pups were divided into three groups, Re-Lenti group (injected with recombinant lentiviral vector, mecp2-sh-1), Lenti group (injected with lentivirus containing no inserted sequences) and a non-injected Control group (Control group). Rats in the injection groups were given intraventricular injections. 7 days and 21 days following injection, no inflammation was observed in the rat brain in the two injected groups, whereas EGFP expression was readily detectable. Sensory-motor reflexes were transiently abnormal in Re-Lenti group, whereas no abnormality was observed in either Control group. Mecp2 mRNA was lower in the hippocampus and cerebral cortex in the Re-Lenti group relative to the Lenti and Control groups. Although no typical RTT like symptoms were observed, the neonatal rats injected with recombinant lentivirus displayed some transient neurobehavioral abnormalities during early development. Bdnf mRNA expression decreased in the hippocampus, supporting the hypothesis that Bdnf may be a target gene of MeCP2 in the CNS. PMID- 18396006 TI - Eating disorders and obsessive-compulsive disorder: A dimensional approach to purported relations. AB - The purpose of this research was to investigate the specificity of purported relations between symptoms of eating disorders (ED) and obsessive-compulsive disorder (OCD). Whereas most research has focused on diagnostic comorbidity or between-groups analyses, this study took a dimensional approach to investigate specific relations among symptoms of anorexia, bulimia, and OCD, as well as panic, depression, and general distress in a student sample (N=465). Results were that all symptoms showed significant zero-order correlations, including all ED OCD pairings. After removing general distress variance, however, none of three OCD scales significantly predicted anorexia; only compulsive washing among OCD scales significantly predicted bulimia. Hierarchical multiple regression demonstrated that panic and depression out-performed OCD in predicting bulimia symptoms. Overall, symptoms of ED and OCD did not show unique relations at the level of core dimensions of each construct. A possible link between bulimia and compulsive washing is worth further study. PMID- 18396007 TI - Symptom severity predicts degree of T cell activation in adult women following childhood maltreatment. AB - Although depression is often associated with a reduction in cellular immune responses, other types of emotional disturbance and psychopathology can activate certain aspects of immunity. Activation markers on T cells, in particular, have been found to be elevated in post-traumatic stress states. However, little is known about the relationship between the severity of PTSD symptoms and the degree of change in T cell phenotypes, or about the potential role of neuroendocrine factors in mediating the association. Twenty-four women with a history of sexual trauma during childhood, including 11 who met diagnostic criteria for PTSD, were compared to 12 age-matched, healthy women without a history of maltreatment. The women provided fasted blood samples for enumeration of cell subsets by immunofluorescence and 24-h urine samples for analysis of catecholamine and cortisol levels. The percent of T cells expressing CD45RA, an early activation marker, was higher in the PTSD diagnosed women, and the levels correlated positively with intrusive symptoms and negatively with avoidant symptoms. These alterations in cell surface markers did not appear to be mediated by norepinephrine (NE) or cortisol, making them a distinctive and independent biomarker of arousal and disturbance in PTSD. PMID- 18396008 TI - Trastuzumb exposure during pregnancy. PMID- 18396009 TI - Morphometric analysis of the AMPA-type neurons in the Deiter's vestibular complex of the chick brain. AB - Chicken (Gallus gallus) brains were used to investigate the typology and the immunolabel pattern for the subunits composing the AMPA-type glutamate receptors (GluR) of hindbrain neurons of the dorsal (dND) and ventral nuclei (vND) of the Deiter's vestibular complex (CD), which is the avian correspondent of the lateral vestibular nucleus (LVN) of mammals. Our results revealed that neurons of both divisions were poor in GluR1. The vND, the GluR2/3+ and GluR4+ label presented no area or neuronal size preference, although most neurons were around 75%. The dND neurons expressing GluR2/3 are primarily around 85%, medium to large-sized 85%, and predominantly 60% located in the medial portion of the rostral pole and in the lateral portion of the caudal pole. The majority of dND neurons containing GluR4 are also around 75%, larger (70% are large and giant), exhibiting a distribution that seems to be complementary to that of GluR2/3+ neurons. This distinct arrangement indicates functional differences into and between the DC nuclei, also signaling that such variation could be attributed to the diverse nature of the subunit composition of the GluRs. Discussion addresses the morphological and functional correlation of the avian DC with the LVN of mammals in addition to the high morphological correspondence, To include these data into the modern comparative approach we propose to adopt a similar nomenclature for the avian divisions dND and vND that could be referred as dLVN and vLVN. PMID- 18396010 TI - Digital imaging, archiving, and structured reporting in pediatric echocardiography: impact on laboratory efficiency and physician communication. AB - BACKGROUND: Digital structured reporting (DSR) is an emerging technology in medical information management. In November of 2006, the pediatric echocardiography laboratory at Children's Hospital of Wisconsin transitioned from transcription-based reporting of results to DSR. METHODS: Transthoracic echocardiograms were reviewed for the study duration and time to report completion for October of 2006 (the last month of phone-based transcription) and January of 2007 (the third month of DSR). For both months, the 5 cardiologists responsible for echocardiography reporting were evaluated for report generation time and the 5 sonographers were evaluated for study duration. Data for physicians and sonographers were analyzed individually, as a group between months, between examination types (brief vs comprehensive), and diagnoses (normal vs abnormal findings). RESULTS: The time to generate a final report to the electronic medical record/auto-fax to the referring physician for all examination types between months decreased significantly from the transcription month to the DSR month (median 23.8 vs 1.2 hours; P =.001). This decrease was consistent among physicians and occurred despite an increase in monthly echocardiography volume of 12% between the study periods. No difference was noted in report generation times between brief and comprehensive transthoracic echocardiography for transcription (P =.220) or DSR (P =.185). Analysis of diagnoses after instituting DSR revealed significantly shorter report generation time for normal examination findings versus abnormal examination findings (0.97 vs 1.5 hours; P =.001). Study duration decreased significantly from the transcription month to the DSR month for all examination types (32 vs 27 minutes; P =.001). For both months, the study duration was longer for abnormal versus normal transthoracic echocardiography findings (transcription: 37 vs 27 minutes; DSR: 32 vs 23 minutes). CONCLUSIONS: DSR is a superior technology for pediatric echocardiography reporting and an important next step to increase efficiency and customer satisfaction. Although congenital heart disease increases DSR study report time, the value of digitally scripted, database-compatible, complete segmental analysis of the heart and vascular structures with direct transfer of quantitative data from the ultrasound machine to the report without the need for a transcription interface justifies the time expense. DSR should be a goal for all pediatric echocardiography laboratories to decrease consumer wait time, increase laboratory efficiency, and improve compliance with Intersocietal Commission for the Accreditation of Echocardiography Laboratories standards. PMID- 18396011 TI - Tricuspid annular and septal Doppler tissue velocities are reduced in pediatric heart transplant recipients without acute rejection. AB - BACKGROUND: We sought to determine any differences between myocardial velocities in healthy children and pediatric heart transplant recipients without acute allograft rejection and to study the clinical factors that could alter these velocities. METHODS: Fifty-eight pediatric heart transplant recipients without rejection and 27 healthy controls were prospectively enrolled. Doppler tissue imaging (DTI) was performed at the tricuspid annulus, septum, and mitral annulus. The influence of the following factors on DTI was assessed: time since transplant, extracorporeal membrane oxygenation, systemic hypertension, graft atherosclerosis, pulmonary hypertension, donor heart ischemic time, and previous rejection. RESULTS: The mean tricuspid annular and septal tissue velocities were significantly reduced in the transplant group compared with controls. The mitral annular velocities were similar in the two groups. Donor heart ischemic time and previous rejection significantly altered DTI velocities. CONCLUSION: Tricuspid annular and septal Doppler tissue velocities are significantly reduced in pediatric heart transplant recipients without allograft rejection and can be altered by prolonged donor heart ischemic time and previous rejection. PMID- 18396012 TI - ERK activation is regulated by E2F1 and is essential for E2F1-induced S phase entry. AB - The E2F family of transcription factors regulates a diverse array of cellular functions including cell cycle progression, cell differentiation and apoptosis. Recent studies indicate that E2F1 influences the activity of signal transduction pathways. We identify here a novel link between E2F1 and the Ras/Raf/MEK/ERK signaling pathway, namely that E2F1 levels affect growth factor-induced ERK phosphorylation. Specifically, downregulating E2F1 inhibits PDGF-induced ERK phosphorylation and ectopic expression of E2F1 sensitizes cells to PDGF. We demonstrate that E2F1 induces ERK activation via a transcriptional mechanism and upregulates the expression of two guanine nucleotide exchange factors, RASGRP1 and RASGEF1B, which promote Ras activation. Furthermore, we show that E2F1 induced ERK activity is essential for E2F1-induced S phase entry. Current literature dictates that the cyclin D/pRB/E2F pathway lies downstream of the mitogenically activated Ras/Raf/MEK/ERK cascade. Our results indicate that the relationship between these signaling modules is not a simple unidirectional linear one and suggests there exists a positive feedback loop that may enhance both ERK signaling and E2F1 activity. PMID- 18396013 TI - Lysophosphatidic acid (LPA) induces the expression of VEGF leading to protection against apoptosis in B-cell derived malignancies. AB - Vascular endothelial growth factor (VEGF) is a survival and angiogenesis factor that is a target for therapy in a variety of cancers. In many hematological malignancies, VEGF production is increased leading to cell survival responses. Herein, we demonstrate that lysophosphatidic acid (LPA) induces mRNA expression of VEGF in the multiple myeloma cell line, U266, the Burkitt's lymphoma cell line, BJAB, and the chronic lymphocytic leukemia (CLL)-like cell line, I-83. This increase in mRNA levels of VEGF corresponded with increased luciferase activity of the VEGF promoter in BJAB and I-83 cells and increased protein levels in I-83 cells. Secretion of VEGF was also increased in these cells following LPA treatment. LPA treatment also caused the activation of both VEGFR1 and VEGFR2. The increase in VEGF expression by LPA is mediated by the activation of c-Jun N terminal Kinase (JNK) and transcription factor NFkappaB since blocking JNK or NFkappaB activation inhibited LPA induced VEGF expression. Furthermore, we have demonstrated that LPA protects cells from apoptosis and blocking activation of both VEGFR1 and VEGFR2 using a VEGF receptor kinase inhibitor prevented LPA survival responses. Knocking down expression of VEGFR1 and inhibiting activation of NFkappaB and JNK also blocked LPA induced protection against apoptosis. Taken together, this indicates that LPA contributes to VEGF production in B cell malignancies leading to cell survival. PMID- 18396014 TI - Sebaceous carcinoma of the parotid gland in children: an immunohistochemical and ploidy study. AB - Sebaceous carcinoma (SC) is a rare malignancy, affecting mainly the periocular glands. To the best of the authors' knowledge, this is the first English-language report of parotid SC affecting children; two cases are presented. Immunohistochemical studies included 29 different antibodies (15 of these were cytokeratins, CKs). For each case, DNA ploidy status was determined using isolated nuclei stained with Feulgen and analysed using a DNA image cytometry system. Most of the tumour cells were positive for CKs AE1/AE3, 34B12, 5 and 7. The CK14 pattern depicted the monolayer of basal cells surrounding the islands of malignant tissue, while the more central sebaceous differentiated cells were negative. Epithelial membrane antigen was strongly positive in the well differentiated cells, while most of the basaloid peripheral cells were negative, and only a few cells were positive for carcinoembryonic antigen. beta catenin, E cadherin and C-erb B2 were expressed by most of the cells including the more differentiated sebaceous cells. Tumour cells were negative for muscle or myoepithelial markers, androgen, oestrogen and progesterone receptors. Both SCs were uniformly diploid, and showed low proliferative indices for p53, Ki-67 and Mcm-2, which is consistent with the good clinical course presented by these patients so far. PMID- 18396015 TI - On the effects of topical synthetic pseudoceramides: comparison of possible keratinocyte toxicities provoked by the pseudoceramides, PC104 and BIO391, and natural ceramides. AB - BACKGROUND: Ceramides (Cer) in the stratum corneum are essential for epidermal permeability barrier function. Thus, topical Cer replacement therapy has been employed to improve barrier function in clinical situations associated with Cer deficiency, e.g., atopic dermatitis. Because of the disadvantages of both natural and skin identical-Cer (central nervous system origins and cost, respectively), synthetic chemical mimics, or pseudoceramides (pseudo-Cer), have been utilized as Cer substitutes. Whereas increased levels of intracellular Cer trigger cell growth inhibition and apoptosis, Cer levels are maintained by metabolic/catabolic pathways protecting cells from Cer-induced apoptosis. However, since the metabolic fates of each pseudo-Cer remain unknown, their widespread deployment in topical agents has raised concern about potential toxicities. OBJECTIVE: We compared the effects of two chemically unrelated commercially available pseudo Cer to exogenous cell-permeant (C2)- or natural (C18)-Cer on cell growth and apoptosis thresholds in cultured human keratinocytes (CHK). METHODS: Cell growth and cell toxicity of CHK exposed to either C2-Cer or pseudo-Cer were assessed by MTT and lactate dehydrogenase release assays. Mitochondrial membrane potential, an indicator of apoptosis, was measured using membrane permeabilized semi-intact keratinocytes exposed C2-Cer, natural-Cer or pseudo-Cer. RESULTS: While the cell permeant-Cer inhibits keratinocyte growth and increases cell toxicity, neither of the pseudo-Cer showed these effects. Decreased mitochondrial membrane potential occurred in CHK incubated with cell-permeant- and natural-Cer, but not pseudo Cer. CONCLUSIONS: Taken together with preclinical safety studies of these pseudo Cer and their widespread use over the counter without evidence of toxicity, these studies provide further assurance about the safety of these pseudo-Cer for topical use. PMID- 18396016 TI - [Prospective epidemiological study of rotavirus gastroenteritis in Europe (REVEAL study). Results in the French area of the study]. AB - PRIMARY OBJECTIVE: To estimate the incidence of acute gastroenteritis (AGE) and rotavirus acute gastroenteritis (RVAGE) in children less than 5 years of age seeking medical care in primary care, emergency department, and hospital settings. SECONDARY OBJECTIVES: To compare the clinical profile of RVAGE and non RVAGE and to describe the distribution of RV serotypes among RVAGE cases. METHODS: A prospective primary care, emergency ward and hospital-based observational study was conducted during 1 year in a selected city of France with 250,000 inhabitants. Children less than 5 years of age presenting with symptoms of AGE were included. Rotavirus was identified using an Elisa test in stools. RESULTS: The estimated annual incidence of RVAGE was 1.56% for AGE and 0.87% for RVAGE in hospital, 5.87% for AGE and 2.65% for RVAGE in emergency-wards, 7.39% for AGE and 1.45% for RVAGE in primary care. Total incidence was 14.82% for AGE and 4.96% for RVAGE among children less than 5 years of age. RVAGE were more clinically severe than the AGE: dehydration (26.8% vs. 14.7%, p<0.0001), vomiting 84.9% vs. 60.9%, p<0.0001), fever (74.3% vs. 44.4%, p<0.0001), lethargy (84.9% vs. 70.2%, p<0.0001). G9 serotype was the most frequent serotype encountered (54.7%) during the study period followed by G3 serotype (33.6%) and G2 serotype (7.9%). CONCLUSION: In this study, RVAGE, caused by serotypes G9 and G3, represented about 1/3 of AGE and were more severe than non-RV AGE with twice as high dehydration rate. These results underline the need for continued promotion on the use of oral rehydration fluids and provide some arguments on the benefits of vaccination against rotavirus and also permanent virological monitoring of circulating serotypes. PMID- 18396017 TI - [Heat stroke and topiramate]. AB - Heatstroke, which is a major disorder related to environmental hyperthermia, is a rare event in children. The risk is increased with predisposing medical conditions and specific medications. We report the case of a 10-year-old epileptic patient, who received topiramate. Topiramate causes hypohydrosis and hyperthermia. We suggest that topiramate treatment may be a risk factor for heatstroke. PMID- 18396018 TI - Impact of obesity on young healthy male adults. PMID- 18396019 TI - Improved formulation of photosensitizer chlorin e6 polyvinylpyrrolidone for fluorescence diagnostic imaging and photodynamic therapy of human cancer. AB - An improved formulation of the photosensitizer chlorin e6 (Ce6) in combination with the hydrophilic polymer polyvinylpyrrolidone (PVP) was investigated for its potential clinical applications in fluorescence diagnosis and photodynamic therapy (PDT) of cancer. This study reports the comparative preclinical biodistribution and efficacy of Ce6 delivered with or without PVP versus dimethyl sulfoxide (DMSO). The safety and fluorescence pharmacokinetics of Ce6-PVP in humans was also accessed. Biodistribution results showed that Ce6-PVP had higher tumor to normal tissue ratio compared to the other formulations. The sensitivity and specificity derived from the area under the receiver operating characteristics curves showed that the formulations were able to discriminate tumor from peritumoral muscle in the following order: Ce6-PVP > Ce6 > Ce6-DMSO. In vitro PDT results showed that Ce6-PVP was found to induce selective phototoxicity in leukemic cells compared to peripheral mononuclear blood cells. In addition, in vivo light irradiation at 1h after Ce6-PVP was found to induce greater tumor necrosis without causing animal toxicity. In patients, preferential accumulation of Ce6-PVP was observed in angiosarcoma lesions compared to normal skin following intravenous administration. In conclusion, PVP significantly enhanced the Ce6 concentration in tumors compared with Ce6 alone and increased the therapeutic index of PDT without any side effects in animal model. No serious adverse events were observed in human as well. PMID- 18396020 TI - Effect of the type of lubricant on the characteristics of orally disintegrating tablets manufactured using the phase transition of sugar alcohol. AB - The aim of this study was to evaluate the effect of lubricants on the characteristics of orally disintegrating (OD) tablets manufactured using the phase transition of sugar alcohol. OD tablets were produced by directly compressing a mixture containing lactose-xylitol granules, disintegrant, glidant and lubricant, and subsequent heating. The effect of the type of lubricant on the tablet characteristics was evaluated using magnesium stearate (Mg-St), sodium stearyl fumarate (SSF), and talc as lubricants. The hardness of the tablets increased to ca. 6kp as a result of heating, regardless of the kind of lubricant. The oral disintegration time of the tablets containing Mg-St or SSF increased with an increase in the hardness. In contrast, the oral disintegration time of the tablets containing talc was not changed despite of an increase in hardness. The water absorption rate of the tablets containing talc was much faster than that of the tablets containing other lubricants. The surface free energy measurement showed that the polarity of the tablet components containing talc was remarkably increased by heating. The water absorption rate of the tablets containing talc was also increased by heating. These results indicate that a more hydrophilic surface might be attained by heating the talc. Consequently, talc was demonstrated to be the most desirable lubricant for the preparation of OD tablets based on the principle of the phase transition of sugar alcohol. PMID- 18396021 TI - Statistical analysis of in vitro data for risk assessment - exemplified for a case of Ames test data. AB - Ames test data of experiments with smoke of six cigarette types were used for dose-response analysis and for derivation of a measure of mutagenic potency. Each cigarette type had been tested using a smoking machine and four dilutions of the smoke of each of seven cycles (one to seven cigarettes). Three plates had been exposed per cigarette number/smoke dilution combination and three control plates had been simultaneously exposed to clean air with each set of smoke-exposed plates. It was the aim of the statistical analysis to determine the slopes of dose-response relationships of various cigarette types and to compare them using statistical tests. Basically, the following procedure is recommended: (1) calculate a dose measure on the basis of the number of smoked cigarettes per cycle and dilution air flow. (2) Use the absolute count values of the individual plates as effect variable. (3) Describe the dose-response relations of the individual cigarette types on the basis of all available data with a polynomial model by means of Poisson regression analysis accounting for overdispersion. (4) Identify the linear dose-response region using the likelihood ratio test and restrict the data set to this region. (5) Use the slope of the linear model in the restricted data set as the basis of the mutagenicity measure. (6) Compare the slope for the individual cigarette type with the slope for a reference cigarette by means of multivariate Poisson regression using the likelihood ratio test and accounting for overdispersion. It is finally recommended to express the mutagenic potency as percentages related to the reference cigarette K2R4F. This type of cigarette was set here equal to 100%; the following values are then obtained for some commercially available cigarette types: type A 25%, type B 90%, type C 119%, type D 13%, type E 59%. The differences are statistically significant. PMID- 18396022 TI - Schlemm, the body snatcher? AB - Friedrich Schlemm (1795-1858) is well known for his original description of the scleral venous sinus, known since as Schlemm's canal. He grew up in a village in the Duchy of Braunschweig (Brunswick). As his family could not afford higher education, he was apprenticed to a barber-surgeon in Braunschweig. This gave him the opportunity to study anatomy and surgery at the local Anatomico-Surgical Institute. Recently discovered archival sources demonstrate that, in June of 1816, Schlemm and a fellow student disinterred the body of a deceased woman late at night in a Braunschweig churchyard to bring the body to this Institute and study the effects of rickets on the woman's bones. They were caught and sentenced to 4 weeks of prison. Subsequently, Schlemm left Braunschweig and found work as a low-rank army surgeon in Berlin. Professor Rudolphi, the director of the Berlin Institute of Anatomy, took note of Schlemm's manual dexterity in anatomical dissection and supported his impressive career. Schlemm eventually became full professor of anatomy in 1833 and spent his remaining 25 years in Berlin with a focus on teaching students and training surgeons. As historical background information is largely lacking in this regard, it is impossible to decide whether Schlemm's episode of grave robbing was a solitary instance or a more common method of acquiring bodies for anatomical instruction in early 19th century Germany. PMID- 18396023 TI - Morphological analysis of the functional design of the connection between the alimentary tract and the gas bladder in air-breathing lepisosteid fish. AB - There is considerable controversy in the literature regarding the existence of a "longitudinal slit" versus a "duct" connecting the respiratory gas bladder with the alimentary tract in various species of garfishes (Lepisosteidae). The aim of the present work has been to address these discrepancies on the basis of our own work on longnose gar Lepisosteus osseus in conjunction with a review of the literature. We provide anatomical evidence for a better understanding of the functional advantages of the "longitudinal slit" versus the "duct" in respect to feeding and respiration in juveniles longnose gar. The anterior part of the garfish body cavity, from the oral cavity to the stomach, was used for morphological and histological analysis. It has been observed that the air bladder in the longnose gar is open to the alimentary tract through a longitudinal slit which is present in the glottis on the dorsal side of the oesophagus. The external side of the glottal ridges in the anterior oesophagus, on the side of the opening, consist of stratified squamous epithelium which then turns into the pseudostratified columnar ciliated epithelium in the posterior oesophagus. The glottal ridges are comprised of epithelia and radially arranged striated muscle fibres surrounded by connective tissue. It can be surmised that the tunica muscularis in the anterior oesophagus of garfish aids the glottis in moving air in and out because it supports the dorsal retractor muscle in propelling air through the longitudinal slit from the buccal cavity. When the buccal cavity of the garfish is contracted during holding prey, the muscles of the gas bladder wall and tunica muscularis, located inside and outside the glottal ridge, participate in exhalation. PMID- 18396024 TI - Effects of two sterol biosynthesis inhibitor fungicides (fenpropimorph and fenhexamid) on the development of an arbuscular mycorrhizal fungus. AB - The effects of different concentrations (0.2, 2, 20, 200mgl(-1)) of two sterol biosynthesis inhibitor (SBI) fungicides, i.e. fenpropimorph and fenhexamid, were evaluated on the spore germination, germ tube elongation, sporulation, and root colonization of Glomus intraradices grown monoxenically in association with transformed carrot roots. The percentage of germinated spores incubated on the SBI fungicides and the length of the germ tubes decreased with increasing concentrations of both fungicides. However, for spore germination this impact was fungistatic rather than fungicidal. Extraradical mycelium architecture and spore production in contact with the SBI fungicides were also strongly impacted at high concentration (20mgl(-1)). Conversely, the colonization of roots developing in the fungicide-free compartment, but interconnected with the extraradical mycelium developing on the SBI fungicides, appeared unaffected. Our results demonstrated that the monoxenic culture system could be used as a standardized, reproducible technique to compare the impacts of different molecules on arbuscular mycorrhizal fungi, and for the initial screening of new candidate molecules before registration. PMID- 18396025 TI - Whey for mass production of Beauveria bassiana and Metarhizium anisopliae. AB - Spore production of Beauveria bassiana and Metarhizium anisopliae was studied in a novel whey-based culture media. Spore yield and viability were determined for two B. bassiana (GHA-726 and CA-603) and two M. anisopliae (CA-1 and IMI 330189) isolates following production in three whey-based systems: solid, liquid, and a diphasic production system. Our study indicated that whey permeate can be used effectively for production of spores of entomopathogenic fungi. However, spore yield and viability were significantly influenced by fungal isolate, whey concentration, and the type of production process used. Under the conditions defined in the present study, spore yields ranging from 1.3x10(9)-10x10(11) spores l(-1) of whey medium could be obtained depending on the strain and production process used. Our study revealed that spores produced by all strains in whey-based solid and liquid media showed between 73-99% viability; germination rates were comparable with those obtained using the standard SDA medium. In the two-stage production process, the viabilities of conidia produced by GHA-726, CA 603, and CA-1 were 35-86, 32-98, and 6-29%, respectively; viability was correlated with whey concentration and isolates. Whey permeate can be used as a growth substrate for mass production of biocontrol fungi. We hypothesize that spore yield and viability could be improved by careful selection of whey content in the medium, incorporation of critical additives and optimization of culture conditions. PMID- 18396026 TI - Nebulisers or spacers for the administration of bronchodilators to those with asthma attending emergency departments? AB - BACKGROUND: Systematic reviews and national guidelines conclude that the nebulised route of administration of bronchodilators has no advantage over the use of a spacer in moderately severe exacerbations of asthma. Whether this recommendation is implemented and whether it might affect use of staff time is unknown. OBJECTIVES: To determine the current method of administration of bronchodilators to those with non-life-threatening asthma attending emergency departments (ED) in London, UK and to monitor the implementation of a new policy to administer bronchodilators by spacers in one ED with a special reference to the time taken by nurses to administer the therapy by two different routes. METHODS: Thirty-five EDs in Greater London were surveyed regarding their current practice. A time and motion study was then undertaken in one department observing nurses administering bronchodilators in the 3 weeks before and 3 weeks after a departmental policy change to favour the use of spacer devices rather than nebulisers. RESULTS: The majority of EDs (94.3%) in Greater London were using the nebulised route of administering bronchodilators to the majority of their adult patients. Spacers were more commonly used for the treatment of children (60.3% of departments using spacers and nebulisers or spacers alone). Over half of the hospitals surveyed (51.4%) were unaware that the British Guidelines on Asthma Management suggested that outcomes were the same and that there were potential advantages in the use of a spacer for both adults and children. Time and motion studies showed that the use of a spacer took no more nursing time than administration of the bronchodilator via a nebuliser; in fact treatment and set up time were considerably lower for spacers. CONCLUSION: Spacer administration of bronchodilators to those with asthma attending EDs utilises less treatment time than use of a nebuliser. A survey of EDs in Greater London has shown that despite guideline conclusions there appears to be little evidence of reduction in use of nebulisers; a fear that use of alternatives might take nurses longer is not supported by this study. PMID- 18396027 TI - Association between C-589T polymorphisms of interleukin-4 gene promoter and asthma: a meta-analysis. AB - BACKGROUND: A number of studies of genetic epidemiology have assessed the association of C-589T (also referred to as C-590T; rs number, 2243250) polymorphisms in the promoter region of interleukin-4 (IL-4) gene with asthma in different populations. However, the results are inconsistent and inconclusive. OBJECTIVES: We performed a meta-analysis of the association between C-589T polymorphisms of IL-4 and asthma with the following objectives: to estimate the magnitude of the gene effect and the possible mode of inheritance. METHODS: A genetic model-free approach was used to perform a meta-analysis. Asthma (atopy status nondefined), nonatopic and atopic asthma subgroups were separately analyzed. Heterogeneity and publication bias were also explored. RESULTS: Our meta-analysis summarized the evidence regarding the association between C-589T polymorphisms in the promoter region of IL-4 gene and asthma. When all asthma groups were pooled, a significant association of increased asthma risk and T allele was found. In subgroup analysis, our results indicated a significant association and a recessive genetic mode of C-589T polymorphisms of IL-4 with atopic asthma. The CC genotype was about 21 percent less likely to have atopic asthma than the genotype CT and TT. However, C-589T polymorphisms were not significantly associated with nonatopic asthma. CONCLUSIONS: This meta-analysis suggests there may be an important effect of single nucleotide polymorphisms (SNPs) in the promoter region of IL-4 gene on the pathogenesis of atopic asthma. This warrants further investigation in larger studies and meta-analysis. PMID- 18396028 TI - Effects of inspiratory muscle thixotropy on the 6-min walk distance in COPD. AB - Thixotropy conditioning of inspiratory muscles at lower lung volumes decreases functional residual capacity (FRC) of following breath cycles with increases in inspiratory capacity. It remains uncertain whether this conditioning would improve exercise tolerance in chronic obstructive pulmonary disease (COPD). Sixteen male stable COPD patients with mild to severe airway obstruction participated. Before the study, all patients completed the 6-min walk test at least twice. The 6-min walk distance (6MWD) was measured after single inspiratory muscle training (IMT) maneuver or without intervention (control) in a randomized cross-over fashion. The 6MWD was also measured after thixotropy conditioning of inspiratory muscles at the maximal expiratory position or without intervention (control). There were no significant differences in the 6MWD after the IMT maneuver (493.2+/-83.7m, P>0.05) versus without intervention (495.7+/-85.9m). The 6MWD after thixotropy conditioning (526.2+/-96.3m, P=0.030) was significantly higher than the 6MWD without intervention (504.3+/-94.1m). The 95% confidence interval of the difference was from 2.6 to 41.2m. Percentage predicted FRC correlated positively with differences in the 6MWD between control and after the thixotropy conditioning maneuver (r=0.78, P=0.007), whereas percentage predicted forced expiratory volume in 1s or the BODE index did not correlate with differences in the 6MWD (P>0.05). Thixotropy conditioning increases self-paced walking distance in patients with COPD. Patients with higher resting FRC benefited more from the conditioning with greater walking distance. PMID- 18396029 TI - Community-acquired methicillin-resistant Staphylococcus aureus pneumonia. AB - Community-acquired methicillin-resistant Staphylococcus aureus is increasingly recognized as an important pathogen causing skin and soft tissue infections. We report a case of severe necrotizing pneumonia caused by community-acquired methicillin-resistant S. aureus in a peripartum woman. This case illustrates that community-acquired methicillin-resistant S. aureus must be considered as a potential pathogen in severe community-acquired pneumonia. PMID- 18396030 TI - Exhaled nitric oxide: independent effects of atopy, smoking, respiratory tract infection, gender and height. AB - Measurement of exhaled nitric oxide is widely used in respiratory research and clinical practice, especially in patients with asthma. However, interpretation is often difficult, due to common interfering factors, and little is known about interactions between factors. We assessed the influences and interactions of factors such as smoking, respiratory tract infections and respiratory allergy concerning exhaled nitric oxide values, with the aim to derive a scheme for adjustment. We studied 897 subjects (514 females, 383 males; mean age+/-standard deviation 34.5+/-13.0 years) with and without respiratory allergy (allergic rhinitis and/or asthma), smoking and respiratory tract infection. Logarithmic nitric oxide levels were described by an additive model comprising respiratory allergy, smoking, respiratory tract infection, gender and height (p0.001 each), without significant interaction terms. Geometric mean was 17.5ppb in a healthy female non smoker of height 170cm, whereby respiratory allergy corresponded to a change by factor 1.50, smoking 0.63, infection 1.24, male gender 1.17, and each 10cm increase (decrease) in height to 1.11 (0.90). Factors were virtually identical when excluding asthma and using the category allergic rhinitis instead of respiratory allergy (n=863). Within each category formed by combinations of these different predictors, the range of residual variation was approximately constant. We conclude that the factors influencing exhaled nitric oxide, which we analyzed, act independently of each other. Thus, circumstances such as smoking and respiratory tract infection do not appear to affect the usefulness of exhaled nitric oxide, provided that appropriate factors for adjustment are applied. PMID- 18396031 TI - The requisite level of theory for the computational design of molecularly imprinted silica xerogels. AB - The present manuscript reports the first application of molecular modelling to the design of molecularly imprinted polymers (MIPs) prepared by alkoxysilane sol gel polymerization. The major goal was to determine the requisite level of theory for the selection of suitable alkoxysilane functional monomers. A comparative study, applied to the design of a MIP for beta-damascenone, involving different levels of theory, basis set superposition error (BSSE) correction and basis set augmentation and also semi-empirical methods, was performed. The computations results suggest that the use of the 3-21G basis set concomitantly with a method for BSSE correction represents a good compromise between theory level and computation time for the successful screening of functional monomers. Additionally, a few selected MIPs and their corresponding non-imprinted congeners (NIPs) were prepared and tested in the role of solid-phase extraction (SPE) sorbents. The confrontation of the computational results with the observed performance and morphological characteristics of the prepared MIPs suggest that besides the strength and type of interactions existing between template and functional monomers other concomitant features, related with the sol-gel process, must also be accounted for so that effective molecular imprinting is achieved in an alkoxysilane xerogel. Nevertheless, since an optimal template-functional monomer interaction is a necessary condition for successful imprinting, the choice of the best monomers is still of the greatest importance and the proposed computational method may constitute an expeditious and reliable screening tool. PMID- 18396032 TI - Microfluidic biochip for chemiluminescent detection of allergen-specific antibodies. AB - Protein microarrays for allergen-specific antibodies detection were integrated in microfluidic chips, with imaging chemiluminescence as the analytical technique. This paper demonstrates the feasibility of miniaturized chemiluminescent ELISA by presenting rapid, reproducible and sensitive detection of protein antibodies using microfluidics. Three different proteins, beta-lactoglobulin, peanut lectin and human IgG were immobilized via a "macromolecules to polydimethylsiloxane elastomer (PDMS) transfer" protocol and used as capturing agent for the detection of specific antibodies. A convenient and reversible procedure was used to bond the PDMS microarray substrate to complimentary SU-8/glass microfluidic reaction chambers. The hydrodynamic behaviours of the three proteins interactions within the micro-chambers were investigated to select the most efficient flowing parameters (come to terms with the assay time and performances). The use of optimized conditions led to the concomitant detection of three specific antibodies at pM level in 300 microL and using 6 min sample incubation time. Finally, sera from allergic patients were assayed using the microfluidic device modified with apple hazelnut and pollen allergen. The results obtained compared favourably with those obtained with the classical Pharmacia CAP system. PMID- 18396033 TI - Implementation of alternating excitation schemes in a biochip-reader for quasi simultaneous multi-color single-molecule detection. AB - We report here the development of a device for single-molecule biochip readout using fast alternating excitation. The technology extends standard imaging cytometry by offering additional color channels in excitation. To enable the study of mobile objects, e.g. actively transported vesicles in living cells or freely diffusing lipids in a lipid bilayer, the frequency of the illumination pulses was chosen high enough to virtually freeze the motion of the biomolecules, as they are shifted through the illuminated area. The synchronization of sample illumination, scanning and line-camera readout yield two quasi-simultaneously recorded images covering the same sample region. Diffraction-limited resolution and high localization precision for point-light sources down to approximately 10 nm was shown by scanning immobilized 100 nm fluorescence latex beads. Ultra sensitivity was demonstrated by imaging single fluorescent streptavidin molecules diffusing in a fluid lipid bilayer. Two-color streptavidin labeled with Cy3 and Cy5 could be easily identified in the two respective excitation channels; high accordance in the dye positions confirms the applicability for colocalization studies of moving objects. Finally, scans of antibody-receptor interactions in large populations of live cells illustrate the feasibility of this method for biochip application. PMID- 18396034 TI - Evolutionary and cellular webs in benzylisoquinoline alkaloid biosynthesis. AB - Alkaloids are a group of approximately 12,000 low molecular weight and nitrogenous secondary metabolites found in 20% of plant species. Their potent biological activity suggests that alkaloids function as defense compounds. Benzylisoquinoline alkaloids (BIAs) are derived from tyrosine and are diversified by an intricate biochemical network of intramolecular coupling, reduction, methylation, hydroxylation, and other reactions to generate the estimated 2500 known structures. Several BIAs are used directly as pharmaceuticals or serve as precursors for the synthesis of semi-synthetic drugs. Plants remain the only economical source for the production of compounds such as morphine and codeine owing to their chemical complexity, which makes de novo synthesis challenging and costly. Much research has been directed toward understanding the biosynthesis of the BIAs and manipulating source plants to increase production of key products and pathway intermediates. However, metabolic engineering experiments often yield unexpected results demonstrating the need for an improved perspective on the biochemistry, regulation, and cell biology of BIA pathways. This review summarizes recent advances in the establishment of predictive metabolic engineering within the context of plant alkaloid biosynthesis. PMID- 18396035 TI - Subanaesthetic ketamine spares postoperative morphine and controls pain better than standard morphine does alone in orthopaedic-oncological patients. AB - BACKGROUND: Postoperative pain in patients with bone and soft tissue cancer is different from that of other surgical patients due to the severity of the pain generated during surgery and because many of them have already been in pain preoperatively. The search for optimal intravenous pharmacologic management for this population is an ongoing one. We conducted a 10-month prospective, randomised, double blind study to compare the effects of a standard morphine dose to a 35%-lower dose plus a subanaesthetic dose of ketamine for postoperative pain control in patients undergoing bone and soft tissue cancer surgery under standardised general anaesthesia. METHODS: After extubation, when objectively awake (>or=5/10 on a 0-10 visual analogue scale (VAS)) and complaining of pain (>or=5/10 VAS), patients were connected to an intravenous patient-controlled analgesia (IV-PCA) device that delivered 1.5 mg morphine/bolus (MO group) or 1 mg morphine+5mg ketamine/bolus (MK group), with a 7 min lockout time. Rescue intramuscular diclofenac 75 mg was available Q4/day. Follow-up lasted 96 h. RESULTS: Fifty-seven patients (24 males, aged 18-74 years) completed the study. Pain scores were lower in the MK group compared to the MO patients, although MO patients (n=29) used 32.9+/-24.9 mg/patient morphine during the first 24 postoperative h compared to 14.6+/-11.4 mg/patient (P<0.05) for the MK patients (n=28). At that time point, 11 MO versus 4 MK patients still required IV-PCA (P<0.05). Diclofenac was also used more in the MO group. All vital signs were similar between the groups. The physiotherapy score was 35% higher for the MK patients (P<0.05). No patient had hallucinations. Postoperative nausea and vomiting rates were higher in the MO group. CONCLUSIONS: The use of subanaesthetic ketamine plus 2/3 the standard dose of morphine following bone and tissue resections results in 1) lower and more stable pain score, 2) approximately 60% morphine sparing effect, 3) a shorter period of postoperative IV-PCA dependence. Such therapy is also associated with better early physical performance. PMID- 18396036 TI - Prognostic versus predictive value of biomarkers in oncology. AB - Numerous options are currently available for tumour typing. This has raised intense interest in the elucidation of prognostic and predictive markers. A prognostic biomarker provides information about the patients overall cancer outcome, regardless of therapy, whilst a predictive biomarker gives information about the effect of a therapeutic intervention. A predictive biomarker can be a target for therapy. Amongst the genes that have proven to be of relevance are well-known markers such as ER, PR and HER2/neu in breast cancer, BCR-ABL fusion protein in chronic myeloid leukaemia, c-KIT mutations in GIST tumours and EGFR1 mutations in NSCLC. Several reasons for the difficult elucidation of new markers will be addressed including the involvement of cellular pathways in tumour biology instead of single genes and interference in disease outcome as a result of anticancer therapies. Future perspectives for the development of prognostic and predictive markers will be given. PMID- 18396037 TI - The potential of anti-vascular endothelial growth factor therapy in metastatic breast cancer: clinical experience with anti-angiogenic agents, focusing on bevacizumab. AB - The importance of angiogenesis in tumour growth and development is well known. Overexpression of vascular endothelial growth factor (VEGF), the key mediator of angiogenesis, is associated with poor prognosis in breast cancer. As a result, several therapeutic agents that inhibit the actions of VEGF or its receptors are currently in development for use in metastatic breast cancer (MBC). This review describes the function of VEGF in normal and tumour angiogenesis, explores the rationale behind the use of anti-VEGF therapy in MBC and details the therapeutic impact of such agents on tumour vasculature. Clinical data from trials of anti VEGF agents in MBC are discussed, with a particular focus on the efficacy and safety of bevacizumab, the anti-VEGF agent at the most advanced stage of development in this tumour type. Future potential uses of bevacizumab in breast cancer are introduced. PMID- 18396038 TI - Antioxidant activity of extracts from leaves and roots of Salvia miltiorrhiza Bunge, S. przewalskii Maxim., and S. verticillata L. AB - Three Salvia species have been studied for antioxidant activity in methanol extracts from roots and leaves. The presence of the polyphenols and tanshinones was screened by HPLC and spectrophotometric assays and related to the antioxidant potential. The antioxidant capacity of the studied species is high, but differences between species and organs have been also revealed. Salvia przewalskii leaf extract was the strongest one in all tests, followed by Salvia miltiorrhiza root and Salvia verticillata leaf. Among the roots, the most active was S. miltiorrhiza extract, followed by S. verticillata. The antioxidant activity correlates to the total polyphenol and, depending on the assay, to the hydroxycinnamic acids content. The high content of tanshinones in both S. miltiorrhiza and S. przewalskii roots is unlikely to contribute to the antioxidant activity. PMID- 18396039 TI - Insecticidal activities of leaf essential oils from Cinnamomum osmophloeum against three mosquito species. AB - The larvicidal activities of leaf essential oils and their constituents from six chemotypes of indigenous cinnamon (Cinnamomum osmophloeum Kaneh.) trees were evaluated against three mosquito species. Results of larvicidal tests demonstrated that the leaf essential oils of cinnamaldehyde type and cinnamaldehyde/cinnamyl acetate type had an excellent inhibitory effect against Aedes albopictus larvae, and their LC(50) values in 24h were 40.8 microg/ml (LC(90)=81.7 microg/ml) and 46.5 microg/ml (LC(90)=83.3 microg/ml), respectively. Results of the 24-h mosquito larvicidal assays also showed that the effective constituents in leaf essential oils were trans-cinnamaldehyde and benzaldehyde and that the LC(50) values of these constituents against A. albopictus larvae were below 50 mug/ml. In addition, cinnamaldehyde type leaf essential oil and trans-cinnamaldehyde have also exhibited great larvicidal performance against Culex quinquefasciatus and Armigeres subalbatus larvae. Comparisons of mosquito larvicidal activity of trans-cinnamaldehyde congeners revealed that alpha-methyl cinnamaldehyde, benzaldehyde, and trans-cinnamaldehyde exhibited strong mosquito larvicidal activity. PMID- 18396040 TI - A high yield multi-method extraction protocol for protein quantification in activated sludge. AB - A multi-method extraction protocol based on mechanical, ionic and hydrophobic methods was investigated on two types of activated sludge samples. Extraction methods were chosen with regards to optimal protein yield without cell disruption. Sonication, EDTA and Tween extraction methods were selected and combined. The total amount of protein released by the multi-method protocol sums up to 191 and 264 mg equiv. BSA/g VSS for the two different sludge samples. Protocol repetition on the same sample showed that protein yield after each successive protocol fitted an exponential curve model. The total amount of extractable proteins was evaluated by model predictions, 423 and 516 mg equiv. BSA/g VSS for the two sludge samples. The multi-method extraction protocol appears relevant for harvesting a representative quantity of proteins from the original sample (45-49%), moreover the multi-method criterion of the protocol also offers a heterogeneous pool of proteins. Thus, further qualitative studies may not be biased by the extraction protocol. PMID- 18396041 TI - Molecular features crucial to the activity of pyrimidine benzamide-based thrombopoietin receptor agonists. AB - The identification of small molecule modulators of biological processes mediated via protein-protein interactions has generally proved to be a challenging endeavor. In the case of the thrombopoietin receptor (TPOr), however, a number of small molecule types have been reported to display biological activity similar to that of the agonist protein TPO. Through a detailed analysis of structure activity relationships, X-ray crystal structures, NMR coupling constants, nuclear Overhauser effects, and computational data, we have determined the agonism inducing conformation of one series of small molecule TPOr agonists. The relationship of this agonism-inducing conformation to that of other series of TPO receptor agonists is discussed. PMID- 18396042 TI - Synthesis of gemcitabine triphosphate (dFdCTP) as a tris(triethylammonium) salt. AB - First synthesis of gemcitabine triphosphate (dFdCTP) as a tris(triethylammonium) salt is reported. PMID- 18396043 TI - Late onset in dysferlinopathy widens the clinical spectrum. AB - LGMD2B, Miyoshi Myopathy and Distal Anterior Compartment Myopathy are caused by mutations in the dysferlin gene (DYSF) leading to progressive muscular weakness and wasting with onset usually within the second or third decade of life. We here present a patient with disease onset at 73 years. The presenting symptom was exercise-induced stiffness of the trunk and proximal leg muscles without major progression over a period of 12 years. Gastrocnemius muscle biopsy revealed dystrophic morphology and biochemical depletion of dysferlin, while sequence analysis revealed compound heterozygous splicing mutations of the dysferlin gene. This case represents the eldest age of onset of dysferlinopathy reported so far and widens the clinical spectrum of this disease. PMID- 18396044 TI - Phenotype and clinical course in a family with a new de novo Twinkle gene mutation. AB - The Twinkle gene product is important for mtDNA replication. Only a few reports have investigated the clinically effect of mutations in this gene. We describe a new de novo mutation (1110C>A) in the PEO1 gene in a mother and her two sons. The mother had progressive ophthalmoplegia, limb weakness, sensory neuropathy, elevated resting plasma lactate, glucose intolerance and impaired VO2max while her sons only had mild ptosis. In accordance with the clinical presentation, abnormal morphological findings in muscle and multiple deletions and depletion of mtDNA in muscle were more pronounced in the proband than in her sons. PMID- 18396045 TI - A novel mitochondrial ND5 (MTND5) gene mutation giving isolated exercise intolerance. AB - We describe a patient with isolated exercise intolerance caused by a new, maternally inherited mutation in mitochondrial DNA. The heteroplasmic T>C transition at position 13271 in MTND5 affects a highly conserved base and segregates with the disease, being present at highest levels in skeletal muscle fibres showing abnormal mitochondrial accumulation. This is the 15th mutation affecting the MTND5 subunit of respiratory chain complex I and confirms this protein as an important site for disease with phenotypes ranging from MELAS and infantile encephalopathies to isolated syndromes affecting a single tissue such as Leber hereditary optic neuropathy and now skeletal muscle. PMID- 18396046 TI - Listeria comet tails: the actin-based motility machinery at work. AB - Listeria monocytogenes is a master of mimicry that uses the host cell actin system both to move within the cytoplasm of infected cells and for cell-to-cell spread. Recent studies of Listeria and similarly acting pathogens have generated leaps in our understanding of the actin-based force producing machinery. This machinery is essential for most motile properties of cells, not least for cell migration. In a minimal configuration, it consists of the Arp2/3-complex, Ena VASP proteins, cofilin, capping protein and a nucleation-promoting factor. In this review, we discuss current models of pseudopodial protrusions and describe how the road to more complex models lies open and is already paved by recent studies using Listeria-based biomimetic motility assays. PMID- 18396048 TI - Modifiable performance domain risk-factors associated with slip-related falls. AB - Falls are a major source of injury in older adults. Many falls occur after slipping. This study determined performance-related factors that both contribute to slip-related falls and that may be effectively and efficiently modified through targeted intervention. Thirty-five young adults and 21 healthy older adults (age: 70.9+/-5.1 years) were slipped in a laboratory using a slippery surface. The biomechanics of the 18 older adults who fell and the 30 younger adults who recovered following slips were analyzed. A set of potentially modifiable variables, initially based on significant between-groups differences, was further analyzed using stepwise discriminant analysis and logistic regression. The discriminant analysis correctly categorized 93.8% of the falls and recoveries based on two variables; the velocity of the slipping foot relative to the velocity of the whole body center of mass (COM), and the lateral placement of the recovery foot relative to the COM. The logistic regression determined the expected change in the odds of a recovery following a slip given a hypothesized intervention-induced improvement of these variables. Decreased velocity of the slipping foot relative to the COM, or decreased lateral placement of the recovery foot relative to the COM to zero, increased the odds of recovery by 17% and 27%, respectively. This suggests that intervention targeted at improving these specific lower extremity control variables following the onset of a slip has the potential to significantly decrease slip-related fall risk. PMID- 18396049 TI - Thermodynamics and fluorescence studies of the interactions of cyclooctapeptides with Hg2+, Pb2+, and Cd2+. AB - The purpose of this work is to characterize the interactions of cyclooctapeptides (CP) containing glutamyl and/or cysteinyl residues with common heavy-metal ions in order to facilitate the design of cyclopeptides as sensors for metal ions. Isothermal titration calorimetry studies show that cyclooctapeptides containing glutamyl and/or cysteinyl residues bind these Hg(2+) and Pb(2+) over Cd(2+) and other common metal ions. Differential binding isotherms, in their interactions with Hg(2+), support a two-binding site model, whereas pertinent interactions with Pb(2+) support a 2:1 stoichiometry, suggesting a CP/Pb(2+)/CP mode of complexation. The cyclooctapeptide containing both glutamyl and cysteinyl residues shows a significant binding affinity for Hg(2+) (K(a)=7.6x10(7)M(-1)), which is both enthalpically and entropically driven. The fluorescence of these cyclooctapeptides showed pronounced fluorescence quenching responses to Hg(2+) over Pd(2+) and Cd(2+). Stern-Volmer analyses of the dependence of fluorescence intensity on Hg(2+) and Pb(2+) are reported. The observed trends are useful for the design of Hg(2+) sensors based on fluorophore-tagged cyclooctapeptides. PMID- 18396047 TI - Itinerant exosomes: emerging roles in cell and tissue polarity. AB - Cells use secreted signals (e.g. chemokines and growth factors) and sophisticated vehicles such as argosomes, cytonemes, tunneling nanotubes and exosomes to relay important information to other cells, often over large distances. Exosomes, 30 100-nm intraluminal vesicles of multivesicular bodies (MVB) released upon exocytic fusion of the MVB with the plasma membrane, are increasingly recognized as a novel mode of cell-independent communication. Exosomes have been shown to function in antigen presentation and tumor metastasis, and in transmitting infectious agents. However, little is known about the biogenesis and function of exosomes in polarized cells. In this review, we discuss new evidence suggesting that exosomes participate in the transport of morphogens and RNA, and thus influence cell polarity and developmental patterning of tissues. PMID- 18396050 TI - 1,5-Disubstituted 1,2,3-triazoles as cis-restricted analogues of combretastatin A 4: Synthesis, molecular modeling and evaluation as cytotoxic agents and inhibitors of tubulin. AB - A series of cis-restricted 1,5-disubstituted 1,2,3-triazole analogues of combretastatin A-4 (1) have been prepared. The triazole 12f, 2-methoxy-5-(1 (3,4,5-trimethoxyphenyl)-1H-1,2,3-triazol-5-yl)aniline, displayed potent cytotoxic activity against several cancer cell lines with IC(50) values in the nanomolar range. The ability of triazoles to inhibit tubulin polymerization has been evaluated, and 12f inhibited tubulin polymerization with IC(50)=4.8microM. Molecular modeling experiments involving 12f and the colchicine binding site of alpha,beta-tubulin showed that the triazole moiety interacts with beta-tubulin via hydrogen bonding with several amino acids. PMID- 18396051 TI - Thioxophosphoranyl aryl- and heteroaryloxiranes as the representants of a new class of metallocarboxypeptidase inhibitors. AB - A novel and potent family of metallocarboxypeptidase inhibitors based on thioxophosphoranyl oxiranes is presented. These compounds bear aryl or heteroaryl substituents with trans-stereochemistry with respect to the phosphorylated group and they have been synthesized by the addition of [bis(diisopropylamino)phosphino](trimethylsilyl)carbene to the corresponding aldehydes and the subsequent thiolation of the phosphine. These oxiranes contain a tetrahedral P atom harboring shielded N,N-groups. The screening of their biological activity as metallocarboxypeptidase inhibitors and some structural studies, as well as full experimental details for the new compounds, is disclosed. Thus, from the analysis of their activity against the prototypical metallocarboxypeptidases A and B (CPA and CPB), we have observed that hydrophobic phosphorylated oxiranes perform better as CPB inhibitors, reaching K(i) values comparable to classical synthetic carboxypeptidase inhibitors. X-ray diffraction analysis revealed that the packing in the structure of one phosphorylated oxirane is mediated mainly by hydrophobic contacts and that the N,N-groups are highly flexible. Consequently, phosphorylated oxiranes might constitute an attractive material for subsequent improvements in the design of novel inhibitors against human proteolytic enzymes with enhanced oral availability. PMID- 18396052 TI - Cytopathic effect of the herpes simplex virus type 1 appears stereologically as early as 4 h after infection of Vero cells. AB - Herpes simplex virus (HSV) is reported to produce marked cytopathogenic effect (CPE) in different cell cultures at least 1-3 days after infection. No stereological data for the CPE of any virus in cultured cells is available in the literature. In this study stereological methods were applied for the first time to estimate cell and nucleus volume of the infected Vero cell line with herpes simplex virus type 1 (HSV-1). Vero cells infected with HSV-1 were studied at 4, 9 and 25 h post-infection and compared to uninfected control cells using stereological methods. As compared with the uninfected control, the mean nucleus and cell volume of the HSV infected cells, increased approximately 53 and approximately 46%, respectively, 4 h post-infection. The methodology can be used to rapidly diagnose CPE changes of the viral infection of cell cultures especially for those viruses which do not produce obvious CPE at early stages. In addition, it is possible to explain the intensity of cytopathic changes like ballooning of infected cells or their nuclei. It means the methods can help us to understand how larger or smaller are the cells (or the nuclei) in comparison with the other groups and statistically analyse these changes at early stages. PMID- 18396053 TI - Calibration of a radionuclide calibrator system as a Bulgarian standard for activity. AB - A commercially available radionuclide calibrator of the type Curiementor 3 (PTW, Freiburg, Germany) was calibrated using activity standard solutions, from the Physikalisch-Technische Bundesanstalt (PTB), of a number of photon-emitting radionuclides, in particular those used in nuclear medicine: (18)F, (51)Cr, (67)Ga, (99m)Tc, (111)In, (123)I, (125)I, (131)I, (186)Re, (201)Tl, and for beta emitting nuclides: (32)P, (89)Sr, (90)Y, (204)Tl. An energy-dependent efficiency curve was established to calculate efficiencies for photon-emitting radionuclides. The differences between experimental and calculated radionuclide efficiencies of the measuring system are of the order of a few percent. The performance of the system was checked for instrument stability, using a long lived (137)Cs source, and for linearity using the decay of a (99m)Tc source. The calibration measurements were carried out with 2g of solution in standard PTB type glass ampoules. The dependence of efficiency with varying solution mass and changing ampoule geometry is studied for some of the radionuclides. For example, geometry correction factors are determined for various solution volumes or ampoules with different wall thickness. The measuring system will enable the National Centre of Metrology (NCM) of Bulgaria to provide calibration services for activity measurements, which are traceable to PTB standards, and to organize national comparisons in order to improve the quality of measurements for nuclear medicine in Bulgarian hospitals. Furthermore, it should enable NCM to declare Calibration and Measurement Capabilities (CMCs) and to submit radioactive solutions to the International Reference System (SIR) at the BIPM in order to compare the results with Key Comparison Reference Values (KCRVs). PMID- 18396054 TI - Purification and activity standardisation of a (166m)Ho solution. AB - As part of a project to use the long-lived (T(1/2)=1200a) (166m)Ho as reference source in its reference ionisation chamber, IRA standardised a commercially acquired solution of this nuclide using the 4pibeta-gamma coincidence and 4pigamma (NaI) methods. The (166m)Ho solution supplied by Isotope Product Laboratories was measured to have about 5% Europium impurities (3% (154)Eu, 0.94% (152)Eu and 0.9% (155)Eu). Holmium had therefore to be separated from europium, and this was carried out by means of ion-exchange chromatography. The holmium fractions were collected without europium contamination: 162h long HPGe gamma measurements indicated no europium impurity (detection limits of 0.01% for (152)Eu and (154)Eu, and 0.03% for (155)Eu). The primary measurement of the purified (166m)Ho solution with the 4pi (PC) beta-gamma coincidence technique was carried out at three gamma energy settings: a window around the 184.4keV peak and gamma thresholds at 121.8 and 637.3keV. The results show very good self consistency, and the activity concentration of the solution was evaluated to be 45.640+/-0.098kBq/g (0.21% with k=1). The activity concentration of this solution was also measured by integral counting with a well-type 5''x5'' NaI(Tl) detector and efficiencies computed by Monte Carlo simulations using the GEANT code. These measurements were mutually consistent, while the resulting weighted average of the 4pi NaI(Tl) method was found to agree within 0.15% with the result of the 4pibeta-gamma coincidence technique. An ampoule of this solution and the measured value of the concentration were submitted to the BIPM as a contribution to the Systeme International de Reference. PMID- 18396055 TI - Molecular characterization of cell death induced by a compatible interaction between Fusarium oxysporum f. sp. linii and flax (Linum usitatissimum) cells. AB - The cellular and molecular events associated with cell death during compatible interaction between Fusarium oxysporum sp. linii and a susceptible flax (Linum usitatissimum) cell suspension are reported here. In order to determine the physiological and molecular sequence of cell death of inoculated cells, reactive oxygen species (ROS) production, mitochondrial potential, lipoxygenase, DNase, protease and caspase-3-like activities, lipid peroxidation and secondary metabolite production were monitored. We also used microscopy, in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) and DNA fragmentation assay. Cell death was associated with specific morphological and biochemical changes that are generally noticed in hypersensitive (incompatible) reaction. An oxidative burst as well as a loss of mitochondrial potential of inoculated cells, an activation of lipoxygenase and lipid peroxidation were noted. Enzyme-mediated nuclear DNA degradation was detectable but oligonucleosomal fragmentation was not observed. Caspase-3-like activity was dramatically increased in inoculated cells. Phenylpropanoid metabolism was also affected as demonstrated by activation of PAL and PCBER gene expressions and reduced soluble lignan and neolignan contents. These results obtained in flax suggest that compatible interaction triggers a cell death sequence sharing a number of common features with the hypersensitive response observed in incompatible interaction and in animal apoptosis. PMID- 18396056 TI - The optimal timing to resect pulmonary metastasis. AB - OBJECTIVE: There is no criterion for the timing of surgical resection of pulmonary metastasis. In this study, we investigated the optimal period for pulmonary metastasectomy. METHODS: Between 2000 and 2005, 68 patients underwent complete pulmonary resection of metastatic cancer. Clinical prognostic factor in multivariate analysis was examined. RESULTS: The interval from pulmonary metastasectomy until subsequent recurrence and the interval from detection of pulmonary metastasis until pulmonary metastasectomy were independent prognostic factors. To investigate the relationship between the two characteristics, the 68 patients were divided into two groups according to the interval from lung metastasectomy until subsequent recurrence. Nineteen patients relapsed within 1 year after pulmonary metastasectomy (group A), while 49 patients did not relapse within 1 year (group B). The interval from detection of pulmonary metastasis until pulmonary metastasectomy was significantly shorter in group A than in group B (2.9 months vs 7.1 months, p=0.01). Based on these results, we divided the patients into two different groups and survival was compared. Significantly shorter survival was observed in the patients who underwent pulmonary metastasectomy within 3 months after detection of pulmonary metastasis (group X, n=35) than in those who underwent the surgery beyond 3 months (group Y, n=33). CONCLUSIONS: There were many cases of early relapse after metastasectomy when the interval from detection of pulmonary metastasis until pulmonary metastasectomy was short. Performing metastasectomy at least three months after detection of pulmonary metastasis may significantly improve the prognosis of patients. PMID- 18396057 TI - Heart rate and pulse pressure at rest are major prognostic markers of early postoperative complications after coronary bypass surgery. AB - OBJECTIVE: There is substantial evidence to consider both heart rate (HR) at rest and pulse pressure (PP) as significant markers of cardiovascular prognosis in the general population. Despite this, neither of these two parameters has been taken into consideration in the design of modern coronary artery bypass risk prediction scores, and little data on their early postoperative prognostic value are currently available. We aimed to assess the predictive value of preoperative HR and PP in the 30-day postoperative period. METHODS: We prospectively enrolled all patients referred to our institution for non-urgent coronary artery bypass grafting. We measured HR on ECG at admittance. Preoperative pulse pressure was obtained by the difference of the mean of three consecutive systolic and diastolic blood pressures. The primary outcome combined the 30-day postoperative mortality, myocardial infarction (new Q-waves on ECG or Troponin-I >20 microg/l) and stroke or transient ischemic attack. The secondary outcome corresponded to clinical events only (stroke or death). Statistical analysis was performed by usual methods. RESULTS: We enrolled 1022 patients (age 66.9+/-9.2 years). Those meeting the primary outcome (n=146) had a significantly higher HR (69.9+/-14.3 bpm vs 64.9+/-13.2 bpm, p<0.0001) and a higher proportion presented a PP >70 mmHg (17.1% vs 10.2%, p<0.03). After adjustments for age, gender, systolic blood pressure, preoperative beta-blocker therapy, left ventricular ejection fraction <0.40, unstable cardiac status, redo surgery, peripheral arterial disease, renal failure, and combined vascular surgery, both HR (OR=1.17 per 10 bpm, p<0.03) and PP >70 mmHg (OR=1.99, p=0.03) remained significant risk predictors. Similar results were found when considering only clinical events. CONCLUSION: This prospective study highlights the usefulness of HR and PP as preoperative risk markers in CABG candidates. PMID- 18396058 TI - Serum leptin levels in community acquired pneumonia (CAP) are related to nutritional status and not to acute phase reaction. AB - To determine whether leptin in patients with CAP acts as a nutritional or as an inflammatory marker and whether leptin plays any role regarding survival, we included 222 patients diagnosed of CAP, 142 men and 80 women, median age 74 years. We did not find significant differences in serum leptin levels between CAP patients and healthy controls, even after adjusting by BMI. Serum leptin levels were directly related with BMI, body fat and muscle mass and inversely related with inflammatory markers, including pro- and anti-inflammatory cytokines. Patients with positive blood cultures showed lower serum leptin and raised inflammatory markers. Although patients who died showed lower values of serum leptin, multivariate analysis showed that the prognostic value of low serum leptin levels depends on impaired nutritional status. In conclusion, we suggest that in CAP patients, leptin does not act as an inflammatory reactant but as a nutritional marker. PMID- 18396059 TI - MALDI tissue profiling of integral membrane proteins from ocular tissues. AB - MALDI tissue profiling and imaging have become valuable tools for rapid, direct analysis of tissues to investigate spatial distributions of proteins, potentially leading to an enhanced understanding of the molecular basis of disease. Sample preparation methods developed to date for these techniques produce protein expression profiles from predominantly hydrophilic, soluble proteins. The ability to obtain information about the spatial distribution of integral membrane proteins is critical to more fully understand their role in physiological processes, including transport, adhesion, and signaling. In this article, a sample preparation method for direct tissue profiling of integral membrane proteins is presented. Spatially resolved profiles for the abundant lens membrane proteins aquaporin 0 (AQP0) and MP20, and the retinal membrane protein opsin, were obtained using this method. MALDI tissue profiling results were validated by analysis of dissected tissue prepared by traditional membrane protein processing methods. Furthermore, direct tissue profiling of lens membrane proteins revealed age related post-translational modifications, as well as a novel modification that had not been detected using conventional tissue homogenization methods. PMID- 18396060 TI - Enzyme inhibitor screening by electrospray mass spectrometry with immobilized enzyme on magnetic silica microspheres. AB - In this study, a novel technique for screening inhibitors by electrospray mass spectrometry (ESI-MS) with immobilized enzyme on magnetic microspheres has been demonstrated. First, the model enzyme acetylcholinesterase (AChE) is immobilized onto the 3-glycidoxypropyltrimethoxysilane (GLYMO)-modified magnetic silica microspheres. AChE activity was monitored by biochemical assay that is based on mixing of AChE immobilized microspheres and model substrate acetylcholine, separating and detecting the product through ESI-MS. Stability of the enzyme immobilized microspheres was investigated. No apparent loss of enzyme activity was observed after fivefold reuse of AChE-immobilized microspheres. The enzyme immobilized bioassay was used to effectively identify AChE inhibitors among two standard samples, huperzine A and huperzine B, and their source herbal Huperzia serrata, all of which were spiked into the substrate. The inhibition was determined by measuring a decrease of product formation using ESI-MS. PMID- 18396061 TI - Dynamic expression of immune response genes in the sea bass, Dicentrarchus labrax, experimentally infected with the monogenean Diplectanum aequans. AB - The sea bass, Dicentrarchus labrax, is one of the most extensively farmed marine fishes in the Mediterranean. Under the high-density condition common in aquaculture, the monogenean gill parasite Diplectanum aequans can cause significant economic losses. This study used real-time quantitative PCR to investigate the dynamic expression of immune response genes in sea bass infected with Diplectanum aequans. The target genes, interleukin-1 (IL-1beta, transforming growth factor (TGF-beta and T-cell receptor (TCR-beta), were studied in the gills and spleen of the sea bass from the first day of infection until thirty days post infection. Our results showed that there was an increase in IL-1beta gene expression in the spleen and gills and in TGF-beta gene expression in the gills of infected fish. These results show that parasitic infection induced a local inflammatory reaction and that reaction was restricted to the site of infection. Finally, the absence of relationship between TCR-beta expression and the parasitic infection suggests that the adaptive immune system is not involved in the response against this parasite. PMID- 18396062 TI - Imaging the human motor system's beta-band synchronization during isometric contraction. AB - Rhythmic synchronization likely subserves interactions among neuronal groups. One of the best studied rhythmic synchronization phenomena in the human nervous system is the beta-band (15-30 Hz) synchronization in the motor system. In this study, we imaged structures across the human brain that are synchronized to the motor system's beta rhythm. We recorded whole-head magnetoencephalograms (MEG) and electromyograms (EMG) of left/right extensor carpi radialis muscle during left/right wrist extension. We analyzed coherence, on the one hand between the EMG and neuronal sources in the brain, and on the other hand between different brain sources, using a spatial filtering approach. Cortico-muscular coherence analysis revealed a spatial maximum of coherence to the muscle in motor cortex contralateral to the muscle in accordance with earlier findings. Moreover, by applying a two-dipole source model, we unveiled significantly coherent clusters of voxels in the ipsilateral cerebellar hemisphere and ipsilateral cerebral motor regions. The spatial pattern of coherence to the right and left arm EMG was roughly mirror reversed across the midline, in agreement with known physiology. Subsequently, we analyzed the brain-wide pattern of beta-band coherence to the motor cortex contralateral to the contracting muscle. This analysis did not reveal any convincing pattern. Because the prior cortico-muscular analysis had demonstrated the expected pattern in our data, this negative finding demonstrates a current limitation of the applied method for cortico-cortical coherence analysis. We conclude that during an isometric muscle contraction, several distributed brain regions form a brain-wide beta-band network for motor control. PMID- 18396063 TI - Motor speech perception modulates the cortical language areas. AB - Traditionally, the left frontal and parietal lobes have been associated with language production while regions in the temporal lobe are seen as crucial for language comprehension. However, recent evidence suggests that the classical language areas constitute an integrated network where each area plays a crucial role both in speech production and perception. We used functional MRI to examine whether observing speech motor movements (without auditory speech) relative to non-speech motor movements preferentially activates the cortical speech areas. Furthermore, we tested whether the activation in these regions was modulated by task difficulty. This dissociates between areas that are actively involved with speech perception from regions that show an obligatory activation in response to speech movements (e.g. areas that automatically activate in preparation for a motoric response). Specifically, we hypothesized that regions involved with decoding oral speech would show increasing activation with increasing difficulty. We found that speech movements preferentially activate the frontal and temporal language areas. In contrast, non-speech movements preferentially activate the parietal region. Degraded speech stimuli increased both frontal and parietal lobe activity but did not differentially excite the temporal region. These findings suggest that the frontal language area plays a role in visual speech perception and highlight the differential roles of the classical speech and language areas in processing others' motor speech movements. PMID- 18396064 TI - Influence of data conflict and molecular phylogeny of major clades in Cimicomorphan true bugs (Insecta: Hemiptera: Heteroptera). AB - Cimicomorpha, which consists of 16 families representing more than 19,400 species, is the largest infraorder in Heteroptera, Insecta. We present the first molecular phylogenetic investigation of family relationships of Cimicomorpha, including 46 taxa from 12 of 16 Cimicomorphan families. Three genes, with a total of 3277 bp of sequence data (nuclear 18S rDNA: 2022 bp, 28S rDNA: 755 bp, and mitochondrial 16S rDNA: 498 bp) were analyzed. Data partitions were analyzed separately and in combination, by employing ML (maximum likelihood), MP (maximum parsimony), and Bayesian methods. As saturation was detected in substitutions of 16S rDNA, influence of data conflict in combined analyses was further explored by three methods: the incongruence length difference (ILD) test, the partitioned Bremer support (PBS), and the partition addition bootstrap alteration approach (PABA). PBS and PABA approaches suggested that 16S rDNA was not very suitable for addressing relationships at this level in Cimicomorpha. Our results also supported the nabid-cimicoid lineage for Cimicoidea proposed by Schuh and Stys [Schuh, R.T., Stys, P., 1991. Phylogenetic analysis of Cimicomorphan family relationships (Heteroptera). J. NY Entomol. Soc. 99 (3), 298-350]. Data incongruence and the utility of the three genes were briefly discussed. PMID- 18396065 TI - A dual-labeling method to quantify unscheduled DNA synthesis in primary cells. AB - Unscheduled DNA synthesis (UDS) has been used as an endpoint for measuring DNA damage in vitro and in vivo. Determination of UDS is regarded as a reliable genotoxicity assay by regulatory agencies including US FDA and EPA. In this study, we have developed an improved UDS assay to detect DNA damage and repair processes upon chemical exposure. We utilized a dual-labeling procedure in which fluorescent DAPI stained nuclei of live cells and [(3)H]thymidine labeled cells undergoing new DNA synthesis. The occurrence of UDS in cells was quantified by either manual nucleus counting or net intensity approaches. This assay was validated by testing known genotoxic compounds 2-acetylaminofluorene (2-AAF), ethylmethanesulfonate (EMS), and N-nitrosodimethylamine (NDL) in primary rat hepatocytes as well as in confluent human mammary epithelial cells. In addition, fluorescent labeling of nuclei DNA helped to distinguish apoptotic cells from non apoptotic cells. Chemical effects on cell functions were also examined by conducting the cytotoxicity assay along with the UDS assay. To conclude, the dual labeling UDS assay offers advantages of reduced subjective bias, increasing sensitivity and reproducibility. The assay is suitable for testing compounds in higher capacity format with much less compound needed. PMID- 18396066 TI - Effect of a ketogenic diet on EEG: analysis of sample entropy. AB - Although ketogenic diet (KD) is an effective alternative therapy for controlling intractable seizures, the anticonvulsant mechanism still remains unclear. Sample entropy (SampEn) provides a generalized measure of regularity in time-series data. To investigate the potential anticonvulsive mechanism of a KD, we analyzed the SampEn of electroencephalography (EEG) data in patients with intractable pediatric epilepsy before and after treatment with a KD. Seventeen pediatric patients with epilepsy who were treated with KD were enrolled in present study. Patients were classified as good responder and poor responder according to therapeutic responsiveness on KD. Thirty segments of 30-s epochs were selected before and after KD from each patient which were subject to SampEn. The KD increased the SampEn in the whole patient population; the SampEn increased significantly in all electrodes in the good responders, but the change in SampEn varied according to the electrode in the poor responders. Before the KD, the good responders had significantly lower SampEn values than the poor responders, but after the KD, SampEn values were higher in the good responders than in the poor responders. KD may have an anticonvulsive effect by decreasing the regularity of the EEG dynamics. PMID- 18396067 TI - The cartilage chondrolytic mechanism of fibronectin fragments involves MAP kinases: comparison of three fragments and native fibronectin. AB - OBJECTIVE: To define the role of mitogen activated protein (MAP) kinases in fibronectin fragment (Fn-f) mediated matrix metalloproteinase (MMP) upregulation and damage to bovine cartilage and to compare activities of three Fn-fs with native fibronectin (Fn), which is inactive in terms of cartilage damage. METHODS: Bovine chondrocytes were cultured with three Fn-fs, an amino-terminal 29-kDa, a gelatin-binding 50-kDa and a central 140-kDa Fn-f or native Fn at concentrations from 0.01 to 1 microM, concentrations lower than those found in osteoarthritis synovial fluids. Lysates were probed for activation of MAP kinases, extracellular signal-regulated kinase 1/2 (ERK1/2), p38 and stress activated protein kinase/c jun N-terminal kinase (SAPK/JNK). Confocal fluorescent microscopy was used to visualize movement of activated kinases. Kinase inhibitors were tested for their abilities to block Fn-f mediated protein upregulation of MMP-3 and MMP-13 and Fn f induced depletion of cartilage proteoglycan (PG) from cultured explants. RESULTS: The 29-kDa, the most potent Fn-f in terms of cartilage damage, enhanced phosphorylation of ERK1/2, p38 and JNK1/2 within a 1-h incubation while the 50 and 140-kDa Fn-fs required up to 4 h for maximal activity and native Fn was only minimally active toward p38 and JNK, but did strongly activate ERK1/2. The activated kinases displayed a distribution toward the nuclear membrane and within the nucleus. MAP kinase inhibitors markedly decreased Fn-f mediated upregulation of MMP-3 or MMP-13 and Fn-f mediated cartilage PG depletion. CONCLUSIONS: These results suggest that Fn-fs upregulate MMP-3 and MMP-13 in bovine chondrocytes through MAP kinases and that kinase inhibitors afford protection against this degenerative pathway. PMID- 18396068 TI - Human-derived probiotic Lactobacillus reuteri demonstrate antimicrobial activities targeting diverse enteric bacterial pathogens. AB - Lactobacillus reuteri is a commensal-derived anaerobic probiotic that resides in the human gastrointestinal tract. L. reuteri converts glycerol into a potent broad-spectrum antimicrobial compound, reuterin, which inhibits the growth of gram-positive and gram-negative bacteria. In this study, we compared four human derived L. reuteri isolates (ATCC 55730, ATCC PTA 6475, ATCC PTA 4659 and ATCC PTA 5289) in their ability to produce reuterin and to inhibit the growth of different enteric pathogens in vitro. Reuterin was produced by each of the four L. reuteri strains and assessed for biological activity. The minimum inhibitory concentration (MIC) of reuterin derived from each strain was determined for the following enteric pathogens: enterohemorrhagic Escherichia coli, enterotoxigenic E. coli, Salmonella enterica, Shigella sonnei and Vibrio cholerae. We also analyzed the relative abilities of L. reuteri to inhibit enteric pathogens in a pathogen overlay assay. The magnitude of reuterin production did not directly correlate with the relative ability of L. reuteri to suppress the proliferation of enteric pathogens. Additional antimicrobial factors may be produced by L. reuteri, and multiple factors may act synergistically with reuterin to inhibit enteric pathogens. PMID- 18396069 TI - Arteriovenous fistula formation using transposed basilic vein: extensive single centre experience. AB - OBJECTIVES: The expanding haemodialysis population has lead to increased requirement for more complex vascular access. The aim of this study is to present the results of an extensive series of brachiobasilic arteriovenous fistulae. METHODS: BBAVF were performed using single-stage vein transposition. A retrospective review of case notes was performed. RESULTS: One hundred and sixty eight BBAVF were created in 144 patients. This was the first access procedure in only 30 cases and the fourth or fifth in 30. At 24h, 165 fistulas (98%) were patent. One hundred and eleven fistulas (66%) were used for haemodialysis and 57 (34%) were never used, of which 39 (23%) were due to fistula failure. The cumulative secondary patency at 1, 2 and 3 years was 66%, 50% and 41% respectively. There were 201 complications in 119 patients (71%), including thrombosis (29%), arm oedema (17%), infection (13%) and arterial steal syndrome (11%). Ten angioplasties and 48 operative procedures were performed for complications. Pre-operative ipsilateral subclavian catheter had been placed in 62 cases (37%) and was associated with poor patency. CONCLUSIONS: BBAVF represents an important option for vascular access with acceptable patency rates, although complication rates remain significant. PMID- 18396070 TI - Increased platelet-monocyte aggregation in male claudicants with the Pl(A1/A2) polymorphism of Gp IIb/IIIa. AB - OBJECTIVES: To investigate the relationship between the Pl(A1/A2) polymorphism and platelet activation and aggregation in patients with Peripheral Arterial Disease (PAD). DESIGN: A prospective single-centre cohort study. METHODS: 45 patients with PAD on aspirin 75mg were recruited and phenotyped/genotyped for the Gp IIb/IIIa Pl(A1/A2) polymorphism. Platelet-Monocyte Aggregation (PMAs) was evaluated using flow-cytometry. RESULTS: The formation of PMAs in the Pl(A2) group was higher but not statistically significant (p=0.17). However, when males were analysed separately, the formation of PMAs was significantly higher in the Pl(A2) group (p=0.0192). No difference was seen in the females. CONCLUSIONS: In this study we show that the Pl(A1/A2) polymorphism primarily affects the aggregation of platelets to monocytes in males. The effect is not observed in females and understanding the mechanism behind this may help elucidate the way the polymorphism alters platelet function in the presence of aspirin. PMID- 18396071 TI - Prospective randomized study of carotid endarterectomy with Fluoropassiv thin wall carotid patch versus venous patch. AB - INTRODUCTION: The practice of carotid endarterectomy (CEA) with patch angioplasty is more effective compared to primary closure. However, the type of patch material remains a controversy. The Fluoropassiv thin wall carotid patch is a polyester patch with an interpenetrating, nanometer-scale, solvent-applied surface modification, based on a biocompatible fluoropolymer. The present pilot study is the first clinical trial evaluating results of CEA with Fluoropassiv versus venous patch. MATERIALS/METHODS: Eighty-seven patients were randomized to 42 Fluoropassiv patching and 45 venous patching. Patients were observed by a vascular surgeon and a neurologist and scanned using duplex ultrasound with a follow-up of 2 years. No patients were lost to follow-up. Restenosis was defined as a Peak Systolic Velocity ratio >2.6, lumen reduction >50%. RESULTS: Perioperative stroke rate was 2.4% in the Fluoropassiv group and 8.9% in the venous group (p=0.02; 1 regressive, 4 non-regressive strokes). Multivariate analysis showed that bilateral carotid stenosis and stroke as indication for CEA were related to perioperative stroke. There was no link between perioperative stroke and patch type after correction for these factors. Patch type had no influence on operation time, clamp time, cranial nerve damage, hypertension, hematoma, infections, time to discharge, or early thromboembolic events. There were no significant differences between the Fluoropassiv and the venous group for cumulative mortality (respectively 4.4 vs 4.8%), patch occlusion (4.8 vs 2.2%), or stroke rate during 2 year follow-up (2.2 vs 2.4%). CONCLUSION: This first clinical study with the Fluoropassiv thin wall carotid patch showed no enhanced thrombogenicity compared to a venous patch. The Fluoropassiv patch is not related to a higher rate of postoperative bleeding events either. PMID- 18396072 TI - The association of peripheral arterial occlusive disease with major coronary events in a mediterranean population with low coronary heart disease incidence. AB - OBJECTIVES: The association of peripheral arterial occlusive disease (PAD) association with major coronary events (MCE) has been well documented, nevertheless data are lacking for populations with a low incidence of coronary heart disease (CHD). We aimed to assess the association of PAD with MCE in a Mediterranean population. DESIGN: Prospective survey of 699 55-74 year-old men representative of an urban district near Barcelona (Spain). METHODS: Baseline cardiovascular risk factors, CHD and PAD (ankle/brachial index<0.9) were recorded. MCE were evaluated during the 5-year follow-up. RESULTS: At recruitment 94 subjects (13.4%) had PAD. During follow-up (mean 69.3 months), 35 (5%) subjects suffered a MCE, of whom 12 had PAD, 9 previous symptomatic CHD and 1 subject both conditions. Higher CHD related mortality (8.6% vs 1.4%; p<0.001) and lower MCE-free survival (78.67% vs 93.26%; p<0.001) was observed for PAD subjects. On Cox regression analysis PAD (RR=3; p=0.003) and previous symptomatic CHD (RR=4.1; p<0.001) were associated independently with MCE during follow-up. CONCLUSIONS: Even in a population with a low incidence of CHD there is a strong relationship between PAD and future MCE. Screening for PAD may improve the selection of patients targeted for cardiovascular risk prevention. PMID- 18396073 TI - Anomalous right subclavian artery and coarctation-related aneurysm repaired with bilateral subclavian-to-carotid transposition and exclusion stent-grafting. AB - INTRODUCTION: Aortic arch anomalies are common; however, the presence of concomitant pathology may present a complex management problem. REPORT: A 42 year old lady with anomalous right subclavian artery was found to have recurrent coarctation of the aorta and an aneurysm related to the previous repair. Management of the aneurysm was complicated by the proximity of subclavian artery origins. Bilateral subclavian-to-carotid transposition was undertaken to preserve antegrade vertebral artery flow, with subsequent exclusion stent-grafting of the aneurysm and coarctation. DISCUSSION: This case illustrates combined surgical and interventional radiological repair to deal with a complex thoracic aortic clinical problem. PMID- 18396074 TI - Combined PET-FDG and USPIO-enhanced MR imaging in patients with symptomatic moderate carotid artery stenosis. AB - INTRODUCTION: PET-FDG and USPIO-enhanced MRI are increasingly being used in depicting carotid atheroma inflammation--a risk factor for the high risk plaque. Their combined use has not been previously reported. REPORT: Two patients presenting with stroke and identified with 50% carotid stenosis on duplex ultrasonography, underwent PET FDG and USPIO-enhanced MR imaging. Results were concordant and complementary suggesting that both techniques reflect similar metabolic processes. DISCUSSION: The selection of patients for carotid revascularisation has largely been based on the severity of luminal stenosis alone. The two imaging modalities, which identify inflammatory activity, may be potential surrogate risk markers in the selection of patients eligible for carotid surgery, if plaque inflammation can be correlated with risk of developing clinical symptoms. PMID- 18396075 TI - Donkey dental anatomy. Part 2: Histological and scanning electron microscopic examinations. AB - Ten normal cheek teeth (CT) were extracted at post mortem from donkeys that died or were euthanased for humane reasons. Decalcified histology was performed on three sections (sub-occlusal, mid-tooth and pre-apical) of each tooth, and undecalcified histology undertaken on sub-occlusal sections of the same teeth. The normal histological anatomy of primary, regular and irregular secondary dentine was found to be similar to that of the horse, with no tertiary dentine present. Undecalcified histology demonstrated the normal enamel histology, including the presence of enamel spindles. Scanning electron microscopy was performed on mid-tooth sections of five maxillary CT, five mandibular CT and two incisors. The ultrastructural anatomy of primary and secondary dentine, and equine enamel types-1, -2 and -3 (as described in horses) were identified in donkey teeth. Histological and ultrastructural donkey dental anatomy was found to be very similar to equine dental anatomy with only a few quantitative differences observed. PMID- 18396076 TI - Donkey dental anatomy. Part 1: Gross and computed axial tomography examinations. AB - Post-mortem examination of 19 donkey skulls showed that donkeys have a greater degree of anisognathia (27% width difference between upper and lower jaws) compared to horses (23%). Teeth (n=108) were collected from 14 skulls and examined grossly and by computed axial tomography (CAT). A greater degree of peripheral enamel infolding was found in mandibular cheek teeth (CT) compared to maxillary CT (P<0.001). A significant increase in peripheral cementum from the apical region to the clinical crown was demonstrated in all CT (P<0.0001). All donkey CT had at least five pulp cavities with six pulp cavities present in the 06s and 11s. A new endodontic numbering system for equid CT has been proposed. A greater occlusal depth of secondary dentine (mm) was present in older donkeys (>16 years) than in the younger (<15 years) donkeys studied. Based on gross and CAT examinations, donkey dental anatomy was shown to be largely similar to that described in horses. PMID- 18396077 TI - Increased sensitivity to heat pain in chronic low blood pressure. AB - While in elevated blood pressure reduced sensitivity to acute pain has been well established, little is known about possible alterations in pain perception within the lower range of blood pressure. In this study, sensitivity to heat pain was assessed in 40 subjects with chronic hypotension (mean blood pressure 96.5/57.7 mmHg) and 40 normotensive control persons (mean blood pressure 121.8/77.2 mmHg). Employing a contact thermode, heat stimuli were applied to the forearm. Pain threshold and tolerance were determined. Participants furthermore rated subjective intensities and unpleasantness of tonic heat stimuli (45.5-47.5 degrees C) on visual analogue scales and in a questionnaire. Possible confounding of sensitivity to heat pain with skin temperature, temperature sensitivity and mood was controlled for. In addition to blood pressure, functional features of the arterial baroreceptor system were related to pain experience. Therefore, estimates for the input on the baroreceptors, as well as baroreflex sensitivity were obtained. Hypotensive individuals exhibited markedly reduced pain threshold and pain tolerance, as well as increased sensory and affective pain experience. The measures related to the baroreceptor system were not associated with pain experience, suggesting that no significant modulation of heat pain occurs through this system. The results of this study complete the findings on hypertension related hypoalgesia and suggest an inverse relationship between blood pressure and pain sensitivity across the whole blood pressure spectrum. Furthermore, increased proneness of hypotensive individuals to clinical pain may be discussed. PMID- 18396078 TI - Fractional volume integration in two-dimensional NMR spectra: CAKE, a Monte Carlo approach. AB - Quantitative information from multi-dimensional NMR experiments can be obtained by peak volume integration. The standard procedure (selection of a region around the chosen peak and addition of all values) is often biased by poor peak definition because of peak overlap. Here we describe a simple method, called CAKE, for volume integration of (partially) overlapping peaks. Assuming the axial symmetry of two-dimensional NMR peaks, as it occurs in NOESY and TOCSY when Lorentz-Gauss transformation of the signals is carried out, CAKE estimates the peak volume by multiplying a volume fraction by a factor R. It represents a proportionality ratio between the total and the fractional volume, which is identified as a slice in an exposed region of the overlapping peaks. The volume fraction is obtained via Monte Carlo Hit-or-Miss technique, which proved to be the most efficient because of the small region and the limited number of points within the selected area. Tests on simulated and experimental peaks, with different degrees of overlap and signal-to-noise ratios, show that CAKE results in improved volume estimates. A main advantage of CAKE is that the volume fraction can be flexibly chosen so as to minimize the effect of overlap, frequently observed in two-dimensional spectra. PMID- 18396079 TI - A double-edged sword: erythropoietin eyed in retinopathy of prematurity. PMID- 18396080 TI - Superior oblique tendon spacer with application of nonabsorbable adjustable suture for treatment of Brown syndrome. AB - PURPOSE: To evaluate the outcomes of a superior oblique tendon spacer procedure using nonabsorbable adjustable sutures in patients with Brown syndrome. METHODS: This noncomparative interventional case series includes 25 eyes of 25 patients with Brown syndrome. In all patients the superior oblique tendon was exposed; two nonabsorbable polyester sutures were placed 4 mm apart, and the tendon was cut. With the use of a slipknot, the cut ends of the tendon were separated 5 to 8 mm. Tendon separation was adjusted intraoperatively according to the exaggerated traction test and indirect ophthalmoscopy. RESULTS: Overall 25 eyes of 25 patients with mean age of 8.00 +/- 4.62 years were operated and followed for a mean period of 13.2 +/- 7.6 months (range, 3 to 30 months). Mean elevation in adduction improved from -3.96 before surgery to -0.67 (p < 0.001); mean hypotropia improved from 11.08(Delta) to 0.32(Delta) (p < 0.001). Two patients developed overcorrection, but recurrence was not observed in any case. The patients continued to improve over the follow-up period. CONCLUSIONS: The adjustable superior oblique tendon suture spacer procedure has favorable results and seems to be technically easier than a silicone expander procedure for Brown syndrome. PMID- 18396081 TI - Botulinum toxin injection with and without electromyographic assistance for treatment of abducens nerve palsy: a pilot study. AB - PURPOSE: To compare the effects and complications of botulinum toxin injection into the medial rectus muscle with and without electromyographic (EMG) assistance for treatment of abducens nerve palsy. METHODS: In a prospective, comparative, interventional case series, botulinum toxin was injected into the medial rectus muscle of 23 patients with esotropia secondary to abducens nerve palsy within 3 months of onset. Patients were randomly assigned to two groups. The injection was given with EMG assistance in the first (EMG group) and without EMG in the second group (no-EMG group). Success was defined as a distance esotropia of no more than 10(Delta) in the primary position. Angle of deviation was recorded preoperatively and at 1 week, 3 months, and 6 months; any postoperative complications and side effects were noted. RESULTS: There were 11 patients in the EMG and 12 patients in the no-EMG group. The ocular deviations were significantly improved in both groups with an overall 6 month success rate of 52.2% (p = 0.008 in the EMG group, p = 0.006 in the no-EMG group). Success rates and mean changes of the angle of deviation were not statistically different between the two groups. Postinjection blepharoptosis was found to be significantly more in the EMG group (p = 0.009). CONCLUSIONS: The effectiveness of botulinum toxin injection is the same with or without EMG assistance into the medial rectus muscle for treatment of abducens nerve palsy. EMG assistance is associated with more blepharoptosis. PMID- 18396082 TI - Transfer of the high-GC cyclohexane carboxylate degradation pathway from Rhodopseudomonas palustris to Escherichia coli for production of biotin. AB - This work demonstrates the transfer of the five-gene cyclohexane carboxylate (CHC) degradation pathway from the high-GC alphaproteobacterium Rhodopseudomonas palustris to Escherichia coli, a gammaproteobacterium. The degradation product of this pathway is pimeloyl-CoA, a key metabolite in E. coli's biotin biosynthetic pathway. This pathway is useful for biotin overproduction in E. coli; however, the expression of GC-rich genes is troublesome in this host. When the native R. palustris CHC degradation pathway is transferred to a DeltabioH pimeloyl-CoA auxotroph of E. coli, it is unable to complement growth in the presence of CHC. To overcome this expression problem we redesigned the operon with decreased GC content and removed stretches of high-GC intergenic DNA which comprise the 5' untranslated region of each gene, replacing these features with shorter low-GC sequences. We show this synthetic construct enables growth of the DeltabioH strain in the presence of CHC. When the synthetic degradation pathway is overexpressed in conjunction with the downstream genes for biotin biosynthesis, we measured significant accumulation of biotin in the growth medium, showing that the pathway transfer is successfully integrated with the host metabolism. PMID- 18396083 TI - Variation of dose distribution of stereotactic radiotherapy for small-volume lung tumors under different respiratory conditions. AB - PURPOSE: To clarify the effects of respiratory condition on dose calculation for stereotactic radiotherapy of small lung tumors. METHODS AND MATERIALS: Computed tomography (CT) data were obtained for nine tumors (diameter, 2.1-3.6cm; mean, 2.7cm) during the stable state, deep expiration, and deep inspiration breath-hold states. Rotational Irradiation with 3 non-coplanar arcs (Rotational Irradiation) and static irradiation with 18 non-coplanar ports (Static Irradiation) using 6-MV photons were evaluated using Fast Fourier Transform (FFT) convolution and Multigrid (MG) superposition algorithms. Dose-volume histograms (DVHs), mean path length (PL) and mean effective path-length (EPL) were calculated. RESULTS: Although the PL was larger for the inspiration state than for the stable state and the expiration state, the EPL was 0.4-0.5cm smaller in the inspiration state than in the expiration state (p=0.01 for Rotational Irradiation; p=0.03 for Static Irradiation). The isocenter dose obtained by the FFT convolution algorithm was 7-12% higher than that obtained with the MG superposition algorithm. A leftward shift of the DVH obtained by MG superposition was noted for the inspiration state compared with the expiration state. CONCLUSIONS: The choice of the proper algorithm is important to accounting for changes in respiration state. Differences in isocenter dose were not large among the respiratory states analyzed. EPL was a little shorter for inspiration than for expiration, although there were larger and reverse trends in path length. A leftward shift of the DVH obtained for the inspiration state when MG superposition was used. PMID- 18396085 TI - Impact of Afghan refugees on the infectious disease profile of Pakistan: beyond economy. PMID- 18396084 TI - Survey of antibiotic use of individuals visiting public healthcare facilities in Indonesia. AB - OBJECTIVES: To estimate the antibiotic use of individuals visiting public healthcare facilities in Indonesia and to identify determinants of use against a background of high resistance rates. METHODS: Patients on admission to hospital (group A), visiting a primary health center (group B), and healthy relatives (group C) were included in the study. A questionnaire on demographic, socioeconomic, and healthcare-related items including health complaints and consumption of antibiotics was used. Logistic regression was performed to determine the co-variables of antibiotic use. RESULTS: Of 2996 individuals interviewed, 486 (16%) had taken an antibiotic. Compared to group C (7% consumption), groups B and A exhibited a three-fold and four-fold higher use of antibiotics, respectively. Respiratory (80%) and gastrointestinal (13%) symptoms were most frequent. Aminopenicillins and tetracyclines accounted for 80% of the prescribed antibiotics. Similar antibiotics were self-medicated (17% of users). Age less than 18 years and health insurance were independent determinants of antibiotic use. Urban provenance, being adult, male, and having no health insurance were independent determinants of self-medication. CONCLUSIONS: In addition to health complaints, other factors determined antibiotic consumption. In view of the likely viral origin of respiratory complaints and the resistance of intestinal pathogens, most antibiotic use was probably unnecessary or ineffective. Future interventions should be directed towards healthcare providers. PMID- 18396086 TI - Evaluation of a structured educational programme for type 2 diabetes patients seen in private practice. AB - AIM: Structured education is necessary in the management of a chronic disease such as diabetes and should be readily offered to patients in different settings. Our aim was to demonstrate the feasibility and advantages of a group education programme for type 2 diabetic patients in a private setting in France. METHODS: A programme of group education for patients with type 2 diabetes was initiated by a multidisciplinary group of volunteer healthcare providers, including general practitioners, specialists in diabetology and non-medical members. All volunteers received one day of training, and physicians were instructed to organize several sessions of group education for the type 2 diabetic patients who regularly attended their practice. The first 427 patients entering the programme were included in the study, and asked to fill in a questionnaire to assess their knowledge, beliefs and behaviours with regard to diabetes. Their physician filled in a medical form. Six months later, the same questionnaire and form were sent for follow-up information. RESULTS: At six months versus baseline, patients exhibited small, but consistent, improvements: (i) fasting glucose 142+/-42 mg/dL (P<0.04) vs 146+/-44 mg/dL (P<0.04); (ii) HbA(1c) 7.41+/-1.26% vs 7.57+/-1.33% (P<0.01); and (iii) all of the main parameters of diabetes self-management recorded in the study. The percentage of patients who inspected their feet at least once a week increased from 67 to 77% (P<0.001). Patients improved their knowledge of the disease and developed a more positive attitude towards their diabetes. CONCLUSION: Our study demonstrates that it is possible to organize educational sessions for diabetic patients in a private-practice setting. At six months, patients receiving these sessions showed benefits in terms of blood glucose control and other important markers of self-management of their disease. PMID- 18396087 TI - Underuse of cardioprotective treatment by the elderly with type 2 diabetes. AB - AIMS: To assess whether elderly patients with type 2 diabetes use a comprehensive cardioprotective regimen (CCR) of antihypertensive, lipid-lowering and antiplatelet drugs in the year following oral antidiabetic drug initiation and, if so, to identify the determinants of such use. METHODS: Using the Quebec Diabetes Surveillance System administrative database, we carried out an inception cohort study of individuals aged 66 years and over who began oral antidiabetic therapy between 1998 and 2002. Those individuals with at least one claim in the year after starting antidiabetic treatment for an antihypertensive, a lipid lowering and an antiplatelet drugs were deemed to be using a CCR. A multivariate logistic regression model was built to identify the characteristics associated with CCR use. RESULTS: Of the 48,505 individuals included in the study, 9912 (20.4%) used a CCR during the year following the first antidiabetic claim. Those more likely to use a CCR were men (odds ratio [OR]: 1.2; 99% confidence intervals [CI]: 1.1-1.3), those who had used an antihypertensive (1.6; 1.4-1.7), lipid lowering (7.4; 6.8-8.0) or antiplatelet (7.3; 6.7-7.9) drug in the year before the first antidiabetic claim and those with a preexisting diagnosis of cardiovascular disease (1.9; 1.8-2.1). The odds of using a CCR increased every year. CONCLUSIONS: CCR use by the elderly with type 2 diabetes in the year following antidiabetic initiation is low, and prior use of individual cardioprotective drugs is a strong predictor of its use. These findings suggest that the treatment of important modifiable risk factors for cardiovascular disease is suboptimal. PMID- 18396088 TI - Training-induced improvement in lipid oxidation in type 2 diabetes mellitus is related to alterations in muscle mitochondrial activity. Effect of endurance training in type 2 diabetes. AB - AIM: We investigated whether or not, in type 2 diabetic (T2D) patients, an individualized training effect on whole-body lipid oxidation would be associated with changes in muscle oxidative capacity. METHODS: Eleven T2D patients participated in the study. Whole-body lipid oxidation during exercise was assessed by indirect calorimetry during graded exercise. Blood samples for measuring blood glucose and free fatty acids during exercise, and muscle oxidative capacity measured from skeletal muscle biopsy (mitochondrial respiration and citrate synthase activity), were investigated in the patients before and after a 10-week individualized training program targeted at LIPOXmax, corresponding to the power at which the highest rate of lipids is oxidized (lipid oxidation at LIPOXmax). RESULTS: Training induced both a shift to a higher-power intensity of LIPOXmax (+9.1+/-4.2W; P<0.05) and an improvement of lipid oxidation at LIPOXmax (+51.27+/-17.93 mg min(-1); P<0.05). The improvement in lipid oxidation was correlated with training-induced improvement in mitochondrial respiration (r=0.78; P<0.01) and citrate synthase activity (r=0.63; P<0.05). CONCLUSION: This study shows that a moderate training protocol targeted at the LIPOXmax in T2D patients improves their ability to oxidize lipids during exercise, and that this improvement is associated with enhanced muscle oxidative capacity. PMID- 18396089 TI - Visual event-related potential changes in multiple system atrophy: delayed N2 latency in selective attention to a color task. AB - Recent studies demonstrated an altered P3 component and prolonged reaction time during the visual discrimination tasks in multiple system atrophy (MSA). In MSA, however, little is known about the N2 component which is known to be closely related to the visual discrimination process. We therefore compared the N2 component as well as the N1 and P3 components in 17 MSA patients with these components in 10 normal controls, by using a visual selective attention task to color or to shape. While the P3 in MSA was significantly delayed in selective attention to shape, the N2 in MSA was significantly delayed in selective attention to color. N1 was normally preserved both in attention to color and in attention to shape. Our electrophysiological results indicate that the color discrimination process during selective attention is impaired in MSA. PMID- 18396090 TI - Effects of mobility and location on food access. AB - Access to healthy food has become an important area of investigation for researchers interested in health disparities and inequalities. The debate about the existence and characteristics of 'food deserts' has increased the interest in food availability and equity in health research. This debate is crucial to an understanding of the factors leading to food security. Research reported here used in-depth interviews with respondents without private transport living within and outside food deserts in Adelaide, South Australia. The respondents came from a variety of households, including single and double parent families, and people living alone. The research found that living in a food desert did not, by itself, impose food access difficulties. Far more important was the access to independent transport to shops. A number of features were identified in this research including reliance on supermarkets, difficulties with public transport, and the provision of government schemes and systems that for some made food shopping much easier. The research suggests that food access problems in Adelaide are not so much the product of geographic distance between home and shop, as the social or welfare networks that allow people to access private transport. PMID- 18396091 TI - Object-based auditory and visual attention. AB - Theories of visual attention argue that attention operates on perceptual objects, and thus that interactions between object formation and selective attention determine how competing sources interfere with perception. In auditory perception, theories of attention are less mature and no comprehensive framework exists to explain how attention influences perceptual abilities. However, the same principles that govern visual perception can explain many seemingly disparate auditory phenomena. In particular, many recent studies of 'informational masking' can be explained by failures of either auditory object formation or auditory object selection. This similarity suggests that the same neural mechanisms control attention and influence perception across different sensory modalities. PMID- 18396092 TI - Plant glycosyl hydrolases and biofuels: a natural marriage. AB - Much of what is currently known about the structure, properties and biochemical activities of glycosyl hydrolases (GHs) has resulted from detailed studies of microbial enzymes. Conversely, such information is sparse in the plant GH literature, where the focus has traditionally been on studying expression and biological function. However, the current resurgence of interest in lignocellulosic biofuels is catalyzing new interest in this field, and recent reports suggest that some plant GH families have more in common with their microbial counterparts than was previously suspected. The repertoires of plant GHs, with their associated catalytic activities and polysaccharide binding affinities, may have valuable applications in modifying plant cell wall architecture and in the development and characterization of new bioenergy feedstocks. PMID- 18396093 TI - The great divide: coordinating cell cycle events during bacterial growth and division. AB - The relationship between events during the bacterial cell cycle has been the subject of frequent debate. While early models proposed a relatively rigid view in which DNA replication was inextricably coupled to attainment of a specific cell mass, and cell division was triggered by the completion of chromosome replication, more recent data suggest these models were oversimplified. Instead, an intricate set of intersecting, and at times opposing, forces coordinate DNA replication, cell division, and cell growth with one another, thereby ensuring the precise spatial and temporal control of cell cycle events. PMID- 18396094 TI - Interaction of bracken-fern extract with vitamin C in human submandibular gland and oral epithelium cell lines. AB - The consumption of bracken-fern (Pteridium aquilinum) as food is associated with a high incidence of cancer in humans and animals. Thus far, the carcinogenic effects of bracken-fern consumption could be related to chromosome aberrations verified in animal and in human peripheral lymphocytes. We tested the in vitro effects of vitamin C (10 and 100 microg/ml) on the reversibility of DNA damage caused by bracken-fern on human submandibular gland (HSG) cells and on oral epithelium cells (OSCC-3) previously exposed to bracken-fern extract. DNA damage (i.e. nuclei with increased levels of DNA migration) was determined by comet assay, cell morphology was evaluated by light microscopy and cellular degeneration was assessed by the acridine orange/ethidium bromide fluorescent dyeing test. Results showed that vitamin C alone did not reduce DNA damage caused by bracken-fern in HSG and OSSC-3 cells. However, at a higher concentration (100 microg/ml), vitamin C induced DNA damage in both cell lines. Moreover, vitamin C (10 and 100 microg/ml) together with bracken-fern extract showed synergistic effects on the frequency of DNA damage in HSG cells. In addition, cells treated with bracken-fern extract or vitamin C alone, or with their association, showed apoptosis morphological features, such as chromatin condensation, cytoplasmic volume loss, changes in membrane symmetry and the appearance of vacuoles; these alterations were observed in both cell lines. These results demonstrate that bracken-fern extract was cytotoxic to HSG and OSCC-3 cells, causing cell death by apoptosis, and that vitamin was not able to revert these effects. PMID- 18396095 TI - Synthesis growth and characterization of L-Valinium Picrate a new nonlinear optical crystal. AB - New nonlinear optical (NLO) single crystal of L-Valinium Picrate (LVP) was grown by slow evaporation solution growth technique at room temperature. The cell parameters of the grown crystal were estimated by single crystal X-ray diffraction analysis. The presence of functional groups was identified from Fourier transform infrared spectrum. The transmission and absorption spectra of this crystal show that the lower cut-off wavelength lies at 380 nm. Thermal analysis was performed to study the thermal stability of the grown crystal. The powder second harmonic generation efficiency of the grown crystal measured by Kurtz technique is 2 times efficient of Potassium dihydrogen orthophosphate (KDP). PMID- 18396096 TI - The laser-induced discoloration of stonework; a comparative study on its origins and remedies. AB - For understanding the phenomena associated with the discoloration observed in some cases of infrared laser cleaned stonework surfaces, a comparative study of three different types and morphologies of pollution encrustation and stone substrates was undertaken. Fragments originating from monuments with historic and/or artistic value, bearing homogeneous thin soiling on Pentelic marble (Athens, Greece), thick encrustation on Hontoria limestone (Burgos, Spain) and compact thin crust on gypsum decorations (Athens, Greece), have been studied on the basis of their composition and origin, together with the conditions that may induce yellowing effects upon their laser cleaning with IR wavelengths. While irradiation in the UV (i.e. at 355 nm) could not effectively remove the encrustations studied, irradiation at 1,064 nm was found efficient to remove all the studied pollution accumulations. Discoloration towards yellow was evident in all cases and at different levels, including the samples with intentional patination layer. To the limit of Raman detection no chemical alterations were detected on the irradiated areas while the presence of yellow polar compounds in all the pollution crusts studied supports the argument that the discoloration of the stone surfaces upon their IR irradiation may be due to the uncovering of existing yellow layers as result of the migration of these compounds inwards to the original stone surface. To correct and/or prevent such undesired coloration the use of IR and UV radiation both in sequential and synchronous mode was considered, with positive results. PMID- 18396097 TI - Monoclinic and triclinic polymorphs of 2-{5,5-dimethyl-3-[2-(2,4,6 trimethoxyphenyl)vinyl]cyclohex-2-enylidene}malononitrile-solid-state linear polarized IR-spectroscopy, DFT calculations and vibrational analysis. AB - The linear-dichroic infrared (IR-LD) spectroscopy of oriented solid samples as suspension in nematic liquid crystal have been carried out for experimental IR band assignment and structural information of 2-{5,5-dimethyl-3-[2-(2,4,6 trimethoxyphenyl)vinyl]cyclohex-2-enylidene} malononitrile polymorphs. The last data have been compared with known crystallographic ones, thus determining the validity of IR-LD spectral conclusions as well as its possibility to determination of Davydov's splitting effect and separation of pairs of maxima corresponding to non-equivalent molecules included in the unit cell of given compound. The experimental structural and spectroscopic data in our case are supported with theoretical DFT ones, obtaining both the electronic structure and vibrational frequencies in gas phase. PMID- 18396098 TI - Natural evolution of carpal tunnel syndrome in untreated patients. AB - OBJECTIVE: To describe the clinical and electrophysiological evolution of a group of patients diagnosed with carpal tunnel syndrome (CTS) who have not undergone any type of treatment (injections, braces or surgery). METHODS: Clinical and electrophysiological monitoring was done consecutively in a series of patients who were admitted with a diagnosis of CTS. The average time for follow-up was 2 years. The Historic and objective Classification scale (HiOb) was used to establish the severity of the disease in the test group, and neuro conductions were done to determine the electrophysiological classification. RESULTS: A total of 132 patients were monitored. In 31 patients (23.4% of the cases), the HiOb classification deteriorated, 28.8% remained stable, and 47.6% showed recovery. In the nerve conduction studies, 10 cases (7.6%) exhibited electrophysiological deterioration, 67.4% remained constant and 25% improved. CONCLUSIONS: A significant percentage of patients with carpal tunnel syndrome had no change in their clinical and electrophysiological condition, while some improved spontaneously without treatment. SIGNIFICANCE: Given that most patients diagnosed with CTS remain stable or even improve over time, a conservative treatment may be all that is needed in a selection of patients with this disorder. PMID- 18396099 TI - Subjective complaints in persons under chronic low-dose exposure to lower polychlorinated biphenyls (PCBs). AB - Polychlorinated biphenyls (PCBs) have been in widespread industrial use in the 1960s and 1970s. Despite a worldwide reduction, environmental exposure remains an issue especially in contaminated buildings. Due to the ubiquitous presence and poor degradation of PCBs, public health concerns continue to exist; however, evidence on the actual health effects of chronic low-dose exposure is scanty. The objective of the present study is an assessment of subjective complaints of exposed subjects in comparison to a non-exposed control group and their inter relation to plasma levels of PCB congeners. The plasma concentrations of PCB congeners were measured in 583 subjects who had worked for an average of 14.7+/ 9.6 years in a contaminated building in Germany, and 205 control subjects working in a non-contaminated building. Subjective complaints were assessed with the 24 item 'Giessen Subjective Complaints List' (GSCL-24). The subjects under chronic low-dose exposure scored significantly higher values on all the GSCL subscales except 'stomach complaints' in comparison to the non-exposed subjects and a 'normal' sample derived from the literature. However, thorough statistical analysis revealed no correlation of symptoms and PCB congener plasma concentration; the scores on the subscale 'exhaustion were even higher in subjects with low PCB concentration. Subjects working in a PCB-contaminated building report more subjective complaints in comparison to non-exposed subjects, but the complaints are not related to current PCB plasma concentrations. PMID- 18396100 TI - An outbreak of foodborne botulism in Taiwan. AB - Botulism is a rare but serious paralytic disease caused by botulinum toxin. We report an outbreak of type B botulism in Taiwan in 2006. There were five cases involved in this outbreak. They present ileus and acute cranial nerve dysfunction including dysphagia and blurred vision. One of them had severe neurologic impairment and required mechanical ventilatory support. No patient received antitoxin administration because of delayed diagnosis. The food specimen revealed positive botulism toxin B. There were no fatalities. Consumption of fermented food was significantly associated with this outbreak. We also reviewed the characteristics of cases with botulism in Taiwan since 1985. PMID- 18396101 TI - Effects of single sessions of low-intensity continuous and moderate-intensity intermittent exercise on blood lipids in the same endurance runners. AB - The factors responsible for the acute effects of exercise on blood lipids are not well known, and there have been few studies comparing different kinds of exercise in the same population. The concentration of blood lipids was evaluated in this study at the end and at post-24h of two 14km/90min single exercise sessions: continuous exercise (CE) at 44.5+/-5.6% VO(2max) and intermittent exercise (IE) at 39-72% VO(2max), in subjects with high levels of aerobic training. Fourteen male athletes (endurance runners) took part in this study and each completed a 24h dietary record. The O(2) uptake and CO(2) production were recorded, and blood lactate and blood lipids were measured. The results showed that triacylglycerols were not modified by any kind of exercise. Total cholesterol was increased at the end of both exercises: 7.04% for CE (p<0.001) and 4.23% for IE (p=0.001). High density lipoprotein cholesterol was increased at the end of IE: 11.38% (p=0.03) and low-density lipoprotein cholesterol was increased only at the end of CE: 7.45% (p=0.006). The increase of lipids for CE was negatively correlated with aerobic fitness indicators (heart rate and %HRmax at lactate threshold), and was positively associated with energy expenditure. For IE, %HRmax and lactate were negatively correlated, and the respiratory exchange ratio was positively correlated, with the lipid increase. We conclude that in trained male athletes, a 14km run in 90min induced different changes of lipid profile if the exercise was done continuously or intermittently, and that in CE the extent of these increases was influenced by aerobic fitness. PMID- 18396102 TI - Skeletomuscular development of genital segments in the dragonfly Anax imperator (Odonata, Aeshnidae) during metamorphosis and its implications for the evolutionary morphology of the insect ovipositor. AB - The skeleton-muscular organisation of abdominal segments 7-9 in female Anax imperator L. (Anisoptera, Aeshnidae) was examined in the stages of ultimate larva, teneral imago, and mature imago, with special emphasis on the transformation of the muscle arrangement. The absence of certain muscles in the genital segments compared to the 7th pre-genital segment was noted on all studied stages. Reductions of certain muscles in adults compared to those in larvae are reported. Some of ovipositor's muscles appear already in larvae. Attachment sites of larval muscles are retained in freshly emerged females concurrently with integument transformations. This situation allows for precise determination of the borders of newly differentiated genital sclerites and, therefore, of the possible origin of certain ovipositor elements in odonates. All changes in the segmental sets of studied abdominal muscles during metamorphosis are tabulated, and displacements of muscles are documented and illustrated. Schematic figures illustrating homologies between the parts of larval and imaginal abdominal sclerites are provided. The origins of the components of the endophytic ovipositor in Odonata as well as their implications for the evolutionary morphology of the insect ovipositor are discussed. PMID- 18396103 TI - From bone marrow to microglia: barriers and avenues. AB - Microglia form a unique population of brain-resident macrophages. Although microglia have been involved in multiple disorders of the central nervous system (CNS), the issue of microglial renewal, under normal or pathological conditions, has been controversial. In mice, results from bone marrow chimera studies indicated that microglia are slowly but continuously replenished by bone marrow derived cells. Moreover, such a microglial turnover was found to be greatly accelerated under multiple neurological conditions. However, recent works questioned the use of irradiation/reconstitution experiments to assess microglial turnover. Based on these different studies, we propose here a re-evaluation of microglia origin(s) in the inflamed CNS. We also discuss the therapeutic perspectives offered by the demonstration of an adult microglial lineage, from bone marrow to brain. PMID- 18396104 TI - The immune privilege of the oral mucosa. AB - Despite high bacterial colonization and frequent allergen contact, acute inflammatory and allergic reactions are rarely seen in the oral mucosa. Therefore we assert that immune tolerance predominates at this site and antigen presenting cells, such as dendritic cells and different T cell subtypes, serve as key players in oral mucosal tolerance induction. In this article we describe the mechanisms that lead to tolerance induced in the oral mucosa and how they differ from tolerance induced in the lower gastrointestinal tract. Furthermore we discuss ways in which novel nonparenteral approaches for immune intervention, such as allergen-specific immunotherapy applied by way of the sublingual route, might be improved to target the tolerogenic properties of the sophisticated oral mucosal immune network. PMID- 18396105 TI - TARDBP mutations in amyotrophic lateral sclerosis with TDP-43 neuropathology: a genetic and histopathological analysis. AB - BACKGROUND: TDP-43 is a major component of the ubiquitinated inclusions that characterise amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) with ubiquitin inclusions (FTLD-U). TDP-43 is an RNA-binding and DNA-binding protein that has many functions and is encoded by the TAR DNA binding protein gene (TARDBP) on chromosome 1. Our aim was to investigate whether TARDBP is a candidate disease gene for familial ALS that is not associated with mutations in superoxide dismutase 1 (SOD1). METHODS: TARDBP was sequenced in 259 patients with ALS, FTLD, or both. We used TaqMan-based SNP genotyping to screen for the identified variants in control groups matched to two kindreds of patients for age and ethnic origin. Additional clinical, genetic, and pathological assessments were made in these two families. FINDINGS: We identified two variants in TARDBP, which would encode Gly290Ala and Gly298Ser forms of TDP-43, in two kindreds with familial ALS. The variants seem to be pathogenic because they co segregated with disease in both families, were absent in controls, and were associated with TDP-43 neuropathology in both members of one of these families for whom CNS tissue was available. INTERPRETATION: The Gly290Ala and Gly298Ser mutations are located in the glycine-rich domain of TDP-43, which regulates gene expression and mediates protein-protein interactions such as those with heterogeneous ribonucleoproteins. Owing to the varied and important cellular functions of TDP-43, these mutations might cause neurodegeneration through both gains and losses of function. The finding of pathogenic mutations in TARDBP implicates TDP-43 as an active mediator of neurodegeneration in TDP-43 proteinopathies, a class of disorder that includes ALS and FTLD-U. FUNDING: National Institutes of Health (AG10124, AG17586, AG005136-22, PO1 AG14382), Department of Veterans Affairs, Friedrich-Baur Stiftung (0017/2007), US Public Health Service, ALS Association, and Fundacio 'la Caixa'. PMID- 18396106 TI - Acute hypertensive response in intracerebral haemorrhage: is treatment safe and helpful? PMID- 18396109 TI - Unexpected right hemisphere language representation identified by the intracarotid amobarbital procedure in right-handed epilepsy surgery candidates. AB - The intracarotid amobarbital procedure (IAP) is used for language lateralization in the presurgical evaluation for intractable epilepsy. Some epilepsy surgical centers forgo IAP for right temporal lobectomies in right-handed patients with no personal/family history of left-handedness, implying that right hemisphere language representation does not occur in these patients. To test this hypothesis, a retrospective analysis was performed on 156 consecutive epilepsy surgery candidates who underwent IAP. Of the 156 candidates, 122 were right handed, and 55 of the 122 demonstrated right hemisphere seizure focus. Right hemisphere language representation was found in 22 of 55 patients, with two demonstrating significant right hemisphere language despite a right hemisphere seizure focus and no family history of left-handedness. Only 1 of 156 patients undergoing IAP experienced permanent neurological complications. Although relatively uncommon, right hemisphere language representation may occur more frequently than complications from cerebral angiography and, therefore, presurgical IAP is recommended for all epilepsy surgery candidates regardless of handedness to minimize the risk of severe language decline. PMID- 18396107 TI - Intensive blood pressure reduction in acute cerebral haemorrhage trial (INTERACT): a randomised pilot trial. AB - BACKGROUND: There is much uncertainty about the effects of early lowering of elevated blood pressure (BP) after acute intracerebral haemorrhage (ICH). Our aim was to assess the safety and efficiency of this treatment, as a run-in phase to a larger trial. METHODS: Patients who had acute spontaneous ICH diagnosed by CT within 6 h of onset, elevated systolic BP (150-220 mm Hg), and no definite indication or contraindication to treatment were randomly assigned to early intensive lowering of BP (target systolic BP 140 mm Hg; n=203) or standard guideline-based management of BP (target systolic BP 180 mm Hg; n=201). The primary efficacy endpoint was proportional change in haematoma volume at 24 h; secondary efficacy outcomes included other measurements of haematoma volume. Safety and clinical outcomes were assessed for up to 90 days. Analysis was by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00226096. FINDINGS: Baseline characteristics of patients were similar between groups, but mean haematoma volumes were smaller in the guideline group (12.7 mL, SD 11.6) than in the intensive group (14.2 mL, SD 14.5). From randomisation to 1 h, mean systolic BP was 153 mm Hg in the intensive group and 167 mm Hg in the guideline group (difference 13.3 mm Hg, 95% CI 8.9-17.6 mm Hg; p<0.0001); from 1 h to 24 h, BP was 146 mm Hg in the intensive group and 157 mm Hg in the guideline group (10.8 mm Hg, 95% CI 7.7-13.9 mm Hg; p<0.0001). Mean proportional haematoma growth was 36.3% in the guideline group and 13.7% in the intensive group (difference 22.6%, 95% CI 0.6-44.5%; p=0.04) at 24 h. After adjustment for initial haematoma volume and time from onset to CT, median haematoma growth differed between the groups with p=0.06; the absolute difference in volume between groups was 1.7 mL (95% CI -0.5 to 3.9, p=0.13). Relative risk of haematoma growth >or=33% or >or=12.5 mL was 36% lower (95% CI 0-59%, p=0.05) in the intensive group than in the guideline group. The absolute risk reduction was 8% (95% CI -1.0 to 17%, p=0.05). Intensive BP-lowering treatment did not alter the risks of adverse events or secondary clinical outcomes at 90 days. INTERPRETATION: Early intensive BP-lowering treatment is clinically feasible, well tolerated, and seems to reduce haematoma growth in ICH. A large randomised trial is needed to define the effects on clinical outcomes across a broad range of patients with ICH. FUNDING: National Health and Medical Research Council of Australia. PMID- 18396108 TI - Antioxidants and free radical scavengers do not consistently delay seizure onset in animal models of acute seizures. AB - A number of herbal compounds with direct antioxidant activity slow the onset, or completely block, the occurrence of seizures. This increase in latency has been proposed to be due to the antioxidant activity. This hypothesis was directly tested by determining the effects of Trolox, a vitamin E analog, vitamin C, melatonin, and alpha-lipoic acid on the latency to acute seizures induced with pilocarpine, kainic acid, or subcutaneous pentylenetetrazol (PTZ) in adult rats. Trolox, vitamin C, and alpha-lipoic acid had significant anticonvulsant activity against pilocarpine, but there were no acute changes in reduced glutathione levels at 15 or 120 minutes. Other than reduced mortality with vitamin C in the PTZ model, none of the antioxidants had a significant effect against PTZ- or kainic acid-induced seizures. The lack of consistent anticonvulsant effect suggests that the antioxidant activity of the herbal preparations cannot account for the delay in seizure onset. PMID- 18396110 TI - Sensitivity and specificity of video alone versus electroencephalography alone for the diagnosis of partial seizures. AB - We examined the usefulness of video versus EEG in isolation for the differentiation of epileptic seizures (ES) from psychogenic nonepileptic events (PNEE). Video-EEG recordings of 43 events in 43 patients (27 with ES and 16 with PNEE) were analyzed by experienced clinical epileptologists/electroencephalographers blinded to the patients' clinical histories. Both the video and EEG were scored independently by the same reader for each event. Relying on video recordings alone, the readers correctly identified ES with a sensitivity of 93% and specificity of 94%. Based on EEG data alone, the readers correctly identified ES with a sensitivity of 89% and specificity of 94%. Semiologically, a gradual evolving buildup of visible symptoms, reaching maximal behavioral intensity within 70 seconds of event onset, was a reliable indicator of ES. No patient with ES demonstrated eye closure at the time of peak behavioral manifestations. Although several additional semiologic features were statistically associated with either ES or PNEE, they were less reliably present and, hence, less clinically useful. Correct categorization of some neurobehavioral events can be made by experienced epileptologists on the basis of video or EEG recordings during an event, without simultaneous review of both provided that the full event is recorded. Home video recordings may represent a useful screening tool for a subset of patients with neurobehavioral events of unclear etiology. PMID- 18396112 TI - Determination of amikacin in cerebrospinal fluid by high-performance liquid chromatography with pulsed electrochemical detection. AB - A highly sensitive and fast reversed-phase liquid chromatographic (LC) method combined with pulsed electrochemical detection (PED) was developed for the direct quantification of the aminoglycoside antibiotic amikacin in cerebrospinal fluid (CSF). The limit of quantification obtained was 0.06 microg/ml and linearity was established over the concentration range 0.06-4.00 microg/ml. The recovery was found to be close to 100%. This method was developed in order to study CSF pharmacokinetics of amikacin in neonates. The narrow therapeutic range calls for monitoring to ensure optimal therapy and to minimize the risk of toxic side effects such as nephro- and ototoxicity, especially in populations like preterm neonates at birth, where the predictability of amikacin clearance is limited. Typical problems to be solved were the low amikacin concentrations and the limited sample volume of CSF. PMID- 18396111 TI - ERCC1/XPF limits L1 retrotransposition. AB - Retrotransposons are currently active in the human and mouse genomes contributing to novel disease mutations and genomic variation via de novo insertions. However, little is known about the interactions of non-long terminal repeat (non-LTR) retrotransposons with the host DNA repair machinery. Based on the model of retrotransposition for the human and mouse LINE-1 element, one likely intermediate is an extension of cDNA that is heterologous to the genomic target, a flap intermediate. To determine whether a human flap endonuclease could recognize and process this potential intermediate, the genetic requirement for the ERCC1/XPF heterodimer during LINE-1 retrotransposition was characterized. Reduction of XPF in human cells increased retrotransposition whereas complementation of ERCC1-deficiency in hamster cells reduced retrotransposition. These results demonstrate for the first time that DNA repair enzymes act to limit non-LTR retrotransposition and may provide insight into the genetic instability phenotypes of ercc1 and xpf individuals. PMID- 18396113 TI - A simple method for obtaining transferrins from human plasma and porcine serum: preparations and properties. AB - A simple method was described for the purification of serum transferrin (Tf) from human plasma and porcine serum with relative high yield and purity. The properties including purity, integrity, immunoreactivity and the receptor-binding ability of the proteins were studied by several assays, comprising spectrometry, SDS-PAGE, HPLC, Western blotting, urea electrophoresis, mass spectrometry and cytometry. Analysis from all the different aspects manifested that the proteins were of high purity. The two kinds of Tfs appeared to be iron-saturated as confirmed by their absorbance spectra and urea-PAGE mobility. The specific spectra of absorption of the two Tfs were both at around 465 nm. The relative molecular weights of human Tf (hTf) and porcine Tf (pTf) were determined by SDS PAGE and further identified by MAIDI-TOF mass spectrometry with a result of 79,707 and 79,258, respectively. Immunoblotting assay showed that pTf could react with the anti-human Tf monoclonal antibody with a less level compared to hTf. FACS assays of their binding activities to Tf receptor-positive cell (K562 cell line) indicated that pTf could be recognized by the hTf receptor and internalized into cells, with a slightly less efficacy than hTf. All special property studies demonstrated that pTf was similar to hTf in physical and chemical characteristics, which gave a hint that pTf could substitute for hTf in some kinds of researches, such as using hTf as a carrier in drug targeting system. PMID- 18396114 TI - Helicobacter pylori infection: a clinical overview. AB - BACKGROUND: Helicobcater pylori colonizes the stomach of more than half of the world's population, and the infection continues to play a key role in the pathogenesis of a number of gastroduodenal diseases. Colonization of the gastric mucosa with Helicobcater pylori results in the development of chronic gastritis in all infected individuals and in a subset of patients chronic gastritis progresses to complications (i.e. ulcer disease, gastric neoplasias, some distinct extragastric disorders). The clinical outcome of the disease is dependent on many variables, including Helicobcater pylori genotype, innate host physiology, genetic predisposition and environmental factors. Helicobcater pylori eradication decreases the incidence of gastroduodenal ulcer and prevents its recurrence. Helicobcater pylori eradication for gastric cancer prevention has been suggested by preclinical research and clinical trials, showing even reversibility of precancerous lesions (atrophic gastritis and intestinal metaplasia) after Helicobcater pylori eradication. AIMS: To review the current literature about H. pylori and its related pathologies. CONCLUSION: At present, several clinical manifestations are recognized to be causally linked to Helicobcater pylori infection, and most of them can be cured by Helicobcater pylori eradication. Besides the relationship of Helicobcater pylori and gastroduodenal diseases, it has been well established that Helicobcater pylori infection is also involved in some extragastrointestinal diseases. PMID- 18396115 TI - Natural history of Helicobacter pylori infection. AB - This report describes the modalities of chronic gastritis induced by Helicobacter pylori infection in different populations. The full gamut of lesions representing the precancerous cascade is very prevalent in populations of low socioeconomic background experiencing very high gastric cancer risk, as seen in the Latin American Andes Mountains. In populations of high socioeconomic standards and high cancer risk, such as Japan and Korea, the precancerous cascade predominates and "early" cancers are also diagnosed frequently. Some reports describe frequent corpus atrophy, not prominent in the former group. The so-called African enigma is seen in populations of low socioeconomic standards, usually living at low altitudes, with high prevalence of infection but low frequency of cancer and precancerous lesions. In populations in transition from high to low cancer risk, duodenal ulcer and antral non-atrophic gastritis are frequently seen. In affluent societies at low risk of cancer, such as Western Europe, Australia and North America, mild non-atrophic gastritis associated with low virulence Helicobacter pylori genotypes predominate. The varied phenotypes of gastritis may reflect secular changes in the ecology of our species. PMID- 18396116 TI - Initial exploration of Ti-Ta, Ti-Ta-Ir and Ti-Ir alloys: Candidate materials for coronary stents. AB - The objective of this study was to explore titanium alloys with increased elastic modulus and improved radiopacity, with a view to utilizing titanium in balloon expandable coronary stents. Ti-50Ta, Ti-45Ta-5Ir and Ti-17Ir alloys were prepared using arc-melting techniques. Microstructural and tensile properties were evaluated in solution-treated conditions for each alloy, and also in aged conditions for the Ti-17Ir. An elastic modulus of 128GPa was recorded for the Ti 17Ir alloy and this high value is attributed to the stiff Ti(3)Ir phase present in the eutectoid structure observed. The mechanical properties recorded, in addition to improved radiopacity, make the Ti-17Ir alloy more suitable for stent applications than commercially available titanium materials. Corrosion resistance and biocompatibility have not been assessed but the noble characteristics of iridium suggest that these aspects will be acceptable. PMID- 18396118 TI - Usefulness of intraoperative 99m Tc-MIBI-guided detection for recurrent sites in secondary hyperparathyroidism: a case-controlled study. AB - PURPOSE: (99m)Tc-Methoxyisobutylisonitrile ((99m)Tc-MIBI)-guided surgery for the detection of abnormal parathyroid glands in primary hyperparathyroidism (1-HPT) has gained popularity as an effective technique. However, in secondary hyperparathyroidism (2-HPT), the efficacy of this method remains controversial, especially for the recurrence sites of 2-HPT. METHODS: (99m)Tc-MIBI-guided surgery was performed for 28 recurrent sites of transplanted parathyroid tissue in 4 patients, and the detection rates of this method were compared with the results of preoperative ultrasound (US) examination and computed tomography (CT) scanning. RESULTS: The results of (99m)Tc-MIBI-guided surgery for regions of recurrence were a sensitivity of 100% (28/28) and an accuracy of 100% (29/29), compared with preoperative US and CT which had a sensitivity of 92.9% (26/28) and 0% (0/28), and an accuracy of 89.7% (26/29) and 0% (0/28), respectively. CONCLUSIONS: Intraoperative (99m)Tc-MIBI-guided surgery can identify recurrent parathyroid tissues of 2-HPT more precisely than preoperative US examination or CT scanning, and makes the surgery easier to perform. PMID- 18396119 TI - The use of the triquetrum as bone graft in total wrist arthrodesis. PMID- 18396117 TI - The design of electrospun PLLA nanofiber scaffolds compatible with serum-free growth of primary motor and sensory neurons. AB - Aligned electrospun nanofibers direct neurite growth and may prove effective for repair throughout the nervous system. Applying nanofiber scaffolds to different nervous system regions will require prior in vitro testing of scaffold designs with specific neuronal and glial cell types. This would be best accomplished using primary neurons in serum-free media; however, such growth on nanofiber substrates has not yet been achieved. Here we report the development of poly(L lactic acid) (PLLA) nanofiber substrates that support serum-free growth of primary motor and sensory neurons at low plating densities. In our study, we first compared materials used to anchor fibers to glass to keep cells submerged and maintain fiber alignment. We found that poly(lactic-co-glycolic acid) (PLGA) anchors fibers to glass and is less toxic to primary neurons than bandage and glue used in other studies. We then designed a substrate produced by electrospinning PLLA nanofibers directly on cover slips pre-coated with PLGA. This substrate retains fiber alignment even when the fiber bundle detaches from the cover slip and keeps cells in the same focal plane. To see if increasing wettability improves motor neuron survival, some fibers were plasma etched before cell plating. Survival on etched fibers was reduced at the lower plating density. Finally, the alignment of neurons grown on this substrate was equal to nanofiber alignment and surpassed the alignment of neurites from explants tested in a previous study. This substrate should facilitate investigating the behavior of many neuronal types on electrospun fibers in serum-free conditions. PMID- 18396120 TI - A rare case of SCC in a young patient with incontinentia pigmenti. PMID- 18396121 TI - Tracheal reconstruction with a prefabricated and double-folded radial forearm free flap. AB - We have described a prefabricated radial forearm free flap containing costal cartilage for tracheal reconstruction. The main advantages of this flap are that it requires only one piece of costal cartilage, simple cartilage trimming, and allows the possible reconstruction of both the internal lining and the external resurfacing when the flap itself is folded. This procedure is easy and its outcome is stable. This flap is an attractive option for the reconstruction of large tracheal and skin defects, even in areas previously exposed to surgery or irradiation. PMID- 18396122 TI - The 'WiMP' formula for local anaesthetic volume calculation. PMID- 18396123 TI - Early response to idursulfase treatment in a 3 year-old boy affected of Hunter syndrome. AB - We present a 3-year-old boy affected with Hunter syndrome. When we first evaluated the patient glycosaminoglycans (GAG) in urine were elevated (94.6 ng/nmol/creatinine); the enzyme activity determined in serum was decreased (3.9 nmol/hxml) and the mutation found was N350H, exon 8. His clinical signs were coarse facial features, hepatomegaly (6 cm), splenomegaly (6 cm), elbow stiffness and hypospadias; dilatation of the perivascular spaces and white matter abnormalities, mitral regurgitation. After two weeks on enzyme replacement therapy (ERT) with idursulfase (IDS), the excretion of GAG was decreased to 36.2 ng/nmol/creatinine and the liver and spleen volumes were reduced to normal limits. He was subsequently noted to have a softer, finer skin, he had no further bouts of bronchitis, and his physical activity improved. This indicates that IDS in young children is well tolerated and that it has several effects which may confer clinical benefits with long-term therapy. PMID- 18396124 TI - Relationships of trabecular bone structure with quantitative ultrasound parameters: in vitro study on human proximal femur using transmission and backscatter measurements. AB - The present study was designed to assess the relationships between QUS parameters and bone density or microarchitecture on samples of human femoral trabecular bone. The normalized slope of the frequency-dependent attenuation (nBUA), the speed of sound (SOS) and the broadband ultrasound backscatter coefficient (BUB) were measured on 37 specimens of pure trabecular bones removed from upper parts of fresh human femurs. Bone mineral density (BMD) was assessed using a clinical scanner. Finally, 8 mm diameter cylindrical cores were extracted from the specimens and their microarchitecture was reconstructed after synchrotron radiation microtomography experiments (isotropic resolution of 10 microm). A large number of microarchitectural parameters were computed, describing morphology, connectivity and geometry of the specimens. BMD correlated with all the microarchitectural parameters and the number of significant correlations was found among the architectural parameters themselves. All QUS parameters were significantly correlated to BMD (R=0.83 for nBUA, R=0.81 for SOS and R=0.69 for BUB) and to microarchitectural parameters (R=-0.79 between nBUA and Tb.Sp, R= 0.81 between SOS and Tb.Sp, R=-0.65 between BUB and BS/BV). Using multivariate model, it was found that microstructural parameters adds 10%, 19%, and 4% to the respective BMD alone contribution for the three variables BUA, SOS and BUB. Moreover, the RMSE was reduced by up to 50% for SOS, by up to 21% for nBUA and up to 11% when adding structural variables to BMD in explaining QUS results. Given the sample, which is not osteoporosis-enriched, the added contribution is quite substantial. The variability of SOS was indeed completely explained by a multivariate model including BMD and independent structural parameters (R(2)=0.94). The inverse problem on the data presented here has been addressed using simple and multiple linear regressions. It was shown that the predictions (in terms of R(2) or RMSE) of microarchitectural parameters was not enhanced when combining 2 or 3 QUS in multiple linear regressions compared to the prediction obtained with one QUS parameter alone. The best model was found for the prediction of Tb.Th() from BUA (R(2)=0.58, RMSE=17 microm). Given the high values of RMSE, these linear models appear of limited clinical value, suggesting that appropriate models have to be derived in order to solve the inverse problem. In this regard, a very interesting multivariate model was found for nBUA and BUB with Tb.Th and Tb.N, in agreement with single scattering theories by random medium. However, the source of residual variability of nBUA and BUB (15% and 45% respectively) remained unexplained. PMID- 18396126 TI - Clinical usefulness of measurement of fibroblast growth factor 23 (FGF23) in hypophosphatemic patients: proposal of diagnostic criteria using FGF23 measurement. AB - Fibroblast growth factor 23 (FGF23) plays important roles in the development of hypophosphatemic diseases such as tumor-induced osteomalacia (TIO) and X-linked hypophosphatemic rickets/osteomalacia (XLH). However, clinical usefulness of measurement of FGF23 has not been established. The objective of this study is to examine the importance of FGF23 measurement in the diagnosis of hypophosphatemic diseases. Biochemical parameters concerning phosphate metabolism were analyzed in a cross-sectional study. 32 patients with TIO, 28 patients with XLH and 16 hypophosphatemic patients with other causes including vitamin D deficiency, Fanconi's syndrome and Cushing's syndrome were studied. In patients with TIO and XLH, FGF23 was above the upper limit of the reference range in most patients irrespective of medical treatment. The lowest FGF23 in these patients was 38.0 pg/ml. FGF23 in hypophosphatemic patients with other causes was undetectable (less than 3 pg/ml) in 12 patients and the highest FGF23 in this group was 23.9 pg/ml. Relationship between phosphate and FGF23 indicated that TIO and XLH are diseases with high FGF23 and hypophosphatemia judged by age-dependent reference ranges for serum phosphate. FGF23 measurement is useful for differential diagnosis of hypophosphatemic diseases caused by excess actions of FGF23 and other etiologies. High FGF23 with low phosphate judged by age-dependent reference ranges for phosphate establishes the diagnosis of diseases caused by excess actions of FGF23. PMID- 18396125 TI - Modulation of extracellular matrix protein phosphorylation alters mineralization in differentiating chick limb-bud mesenchymal cell micromass cultures. AB - Protein phosphorylation and dephosphorylation are important regulators of cellular and extracellular events. The purpose of this study was to define how these events regulate cartilage matrix calcification in a cell culture system that mimics endochondral ossification. The presence of casein kinase II (CK2), an enzyme known to phosphorylate matrix proteins, was confirmed by immunohistochemistry. The importance of phosphoprotein phosphorylation and dephosphorylation was examined by comparing effects of inhibiting CK2 or phosphoprotein phosphatases on mineral accretion relative to untreated mineralizing controls. Specific inhibitors were added to differentiating chick limb-bud mesenchymal cell micromass cultures during the development of a mineralized matrix at the times of cell differentiation, proliferation, formation of the mineralized matrix, or proliferation of the mineral crystals. The mineralizing media for these cultures contained 4 mM inorganic phosphate and no organic-phosphate esters; control cultures had 1 mM inorganic phosphate. Mineralization was monitored based on (45)Ca uptake and infrared characterization of the mineral; cell viability was assessed by three independent methods. Treatments that caused cell toxicity were excluded from the analysis. Inhibition of CK2 activity with apigenin or CK2 inhibitor II reduced the rate of mineral deposition, but did not block mineral accretion. Effects were greatest during the time of mineralized matrix formation. Inhibition of phosphoprotein phosphatase activities with okadaic acid, calyculin A, and microcystin-LR, at early time points also markedly inhibited mineral accretion. Inhibition after mineralization had commenced increased the mineral yield. Levamisole, an alkaline phosphatase inhibitor, had no effect on mineral accretion in this system, suggesting the involvement of other phosphatases. Adding additional inorganic phosphate to the inhibited cultures after mineralization had started, but not earlier, reversed the inhibition indicating that the phosphatases were, in part, providing a source of inorganic phosphate. To characterize the roles of specific phosphoproteins blocking studies were performed. Blocking with anti-osteopontin antibody confirmed osteopontin's previously reported role as a mineralization inhibitor. Blocking antibodies to bone sialoprotein added from day 9 or on days 9 and 11 retarded mineralization, supporting its role as a mineralization nucleator. Antibodies to osteonectin slightly stimulated early mineralization, but had no effect after the time that initial mineral deposition occurs. Taken together, the results of this study demonstrate the importance of the phosphorylation state of extracellular matrix proteins in regulating mineralization in this culture system. PMID- 18396127 TI - TIEG-null mice display an osteopenic gender-specific phenotype. AB - TGFbeta inducible early gene-1 (TIEG) was originally cloned from human osteoblasts (OB) and has been shown to play an important role in TGFbeta/Smad signaling, regulation of gene expression and OB growth and differentiation. To better understand the biological role of TIEG in the skeleton, we have generated congenic TIEG-null (TIEG(-/-)) mice in a pure C57BL/6 background. Through the use of DXA and pQCT analysis, we have demonstrated that the femurs and tibias of two month-old female TIEG(-/-) mice display significant decreases in total bone mineral content, density, and area relative to wild-type (WT) littermates. However, no differences were observed for any of these bone parameters in male mice. Further characterization of the bone phenotype of female TIEG(-/-) mice involved mechanical 3-point bending tests, micro-CT, and histomorphometric analyses of bone. The 3-point bending tests revealed that the femurs of female TIEG(-/-) mice have reduced strength with increased flexibility compared to WT littermates. Micro-CT analysis of femurs of two-month-old female TIEG(-/-) mice revealed significant decreases in cortical bone parameters compared to WT littermates. Histomorphometric evaluation of the distal femur revealed that female TIEG(-/-) mice also display a 31% decrease in cancellous bone area, which is primarily due to a decrease in trabecular number. At the cellular level, female TIEG(-/-) mice exhibit a 42% reduction in bone formation rate which is almost entirely due to a reduction in double labeled perimeter. Differences in mineral apposition rate were not detected between WT and TIEG(-/-) mice. Taken together, these findings suggest that female TIEG(-/-) mice are osteopenic mainly due to a decrease in the total number of functional/mature OBs. PMID- 18396128 TI - Comparisons of diets used in animal models of high-fat feeding. PMID- 18396129 TI - Complex I: a complex gateway to the powerhouse. AB - Complex I, the main entry point for electrons to the respiratory chain, is of critical importance for cellular energy homeostasis. In this issue of Cell Metabolism, Kruse and coworkers (2008) describe the first mouse knockout for a complex I structural subunit, thus advancing our understanding of complex I in disease. PMID- 18396130 TI - When BAD is good for beta cells. AB - BAD, a proapoptotic member of the Bcl-2 family of proteins, is regulated by phosphorylation. A recent study (Danial et al., 2008) suggests a phosphorylation state-dependent bifunctional role of BAD in the regulation of glucose-stimulated insulin secretion and beta cell mass. PMID- 18396131 TI - Fly-let biology and the high protein/low carb diet. AB - In Drosophila, a simple network of nutrient-sensing neuroendocrine cells, analogs of pancreatic islet alpha and beta cells, regulates carbohydrate metabolism. Work presented in this issue of Cell Metabolism (Buch et al., 2008) shows that signals from these cells control expression of a glycogen-specific glucosidase in response to dietary protein and carbohydrate. PMID- 18396132 TI - Bruton and Tec: new links in osteoimmunology. AB - Enhanced and deficient immune responses are associated with abnormal bone homeostasis. A new study by Shinohara et al. (2008) shows that protein phosphorylation by the tyrosine kinases Bruton and Tec links immunity and bone as well as two signaling pathways in precursors of osteoclasts, the cells that degrade bone. PMID- 18396133 TI - Diabetes and cognitive decline: are steroids the missing link? AB - Although elevated cortisol levels may be found in diabetes, cortisol's role in diabetes-associated cognitive impairment is unclear. A recent study (Stranahan et al., 2008) shows that reducing glucocorticoid levels in rodent diabetes models prevents cognitive deficits and enhances neurogenesis and synaptic plasticity. The clinical relevance of these intriguing findings remains undetermined. PMID- 18396134 TI - Iron homeostasis: fitting the puzzle pieces together. AB - Here, recent insights into iron homeostasis are highlighted. Three studies demonstrate the importance of the IRE-IRP system for enterocytes in balancing extracellular iron demand against cellular iron requirements, show that the hemochromatosis protein HFE exerts its iron-regulatory activity principally in hepatocytes by modulating the production of hepcidin, and provide strong support for a proposed mechanism of transcriptional regulation of hepcidin through a signaling cascade initiated by holotransferrin displacing HFE from transferrin receptor 1. PMID- 18396135 TI - PDK1 deficiency in POMC-expressing cells reveals FOXO1-dependent and -independent pathways in control of energy homeostasis and stress response. AB - Insulin- and leptin-stimulated phosphatidylinositol-3 kinase (PI3K) activation has been demonstrated to play a critical role in central control of energy homeostasis. To delineate the importance of pathways downstream of PI3K specifically in pro-opiomelanocortin (POMC) cell regulation, we have generated mice with selective inactivation of 3-phosphoinositide-dependent protein kinase 1 (PDK1) in POMC-expressing cells (PDK1(DeltaPOMC) mice). PDK1(DeltaPOMC) mice initially display hyperphagia, increased body weight, and impaired glucose metabolism caused by reduced hypothalamic POMC expression. On the other hand, PDK1(DeltaPOMC) mice exhibit progressive, severe hypocortisolism caused by loss of POMC-expressing corticotrophs in the pituitary. Expression of a dominant negative mutant of FOXO1 specifically in POMC cells is sufficient to ameliorate positive energy balance in PDK1(DeltaPOMC) mice but cannot restore regular pituitary function. These results reveal important but differential roles for PDK1 signaling in hypothalamic and pituitary POMC cells in the control of energy homeostasis and stress response. PMID- 18396136 TI - Hepatic steatosis in leptin-deficient mice is promoted by the PPARgamma target gene Fsp27. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is induced in leptin deficient (ob/ob) mouse liver and is critical for the development of hepatic steatosis. The present study shows that fat-specific protein 27 (Fsp27) in ob/ob liver is a direct target gene of PPARgamma and can elevate hepatic triglyceride levels. FSP27 belongs to the CIDE family, composed of CIDE A, CIDE B, and FSP27/CIDE C, all of which contain a conserved CIDE-N domain. FSP27 was recently reported to be a lipid droplet-binding protein and to promote lipid accumulation in adipocytes. The Fsp27 gene was expressed at high levels in ob/ob liver and at markedly lower levels in ob/ob livers lacking PPARgamma. Forced expression of FSP27 by adenovirus in hepatocytes in vitro or in vivo led to increased triglyceride levels. Knockdown by adenovirus expressing FSP27 shRNA resulted in lower accumulation of hepatic triglycerides compared to control adenovirus infected liver. Taken together, these results indicate that FSP27 is a direct mediator of PPARgamma-dependent hepatic steatosis. PMID- 18396137 TI - Mice with mitochondrial complex I deficiency develop a fatal encephalomyopathy. AB - To study effects of mitochondrial complex I (CI, NADH:ubiquinone oxidoreductase) deficiency, we inactivated the Ndufs4 gene, which encodes an 18 kDa subunit of the 45-protein CI complex. Although small, Ndufs4 knockout (KO) mice appeared healthy until approximately 5 weeks of age, when ataxic signs began, progressing to death at approximately 7 weeks. KO mice manifested encephalomyopathy including a retarded growth rate, lethargy, loss of motor skill, blindness, and elevated serum lactate. CI activity in submitochondrial particles from KO mice was undetectable by spectrophotometric assays. However, CI-driven oxygen consumption by intact tissue was about half that of controls. Native gel electrophoresis revealed reduced levels of intact CI. These data suggest that CI fails to assemble properly or is unstable without NDUFS4. KO muscle has normal morphology but low NADH dehydrogenase activity and subsarcolemmal aggregates of mitochondria. Nonetheless, total oxygen consumption and muscle ATP and phosphocreatine concentrations measured in vivo were within normal parameters. PMID- 18396138 TI - Opposing effects of dietary protein and sugar regulate a transcriptional target of Drosophila insulin-like peptide signaling. AB - Specific neurosecretory cells of the Drosophila brain express insulin-like peptides (dilps), which regulate growth, glucose homeostasis, and aging. Through microarray analysis of flies in which the insulin-producing cells (IPCs) were ablated, we identified a target gene, target of brain insulin (tobi), that encodes an evolutionarily conserved alpha-glucosidase. Flies with lowered tobi levels are viable, whereas tobi overexpression causes severe growth defects and a decrease in body glycogen. Interestingly, tobi expression is increased by dietary protein and decreased by dietary sugar. This pattern is reminiscent of mammalian glucagon secretion, which is increased by protein intake and decreased by sugar intake, suggesting that tobi is regulated by a glucagon analog. tobi expression is also eliminated upon ablation of neuroendocrine cells that produce adipokinetic hormone (AKH), an analog of glucagon. tobi is thus a target of the insulin- and glucagon-like signaling system that responds oppositely to dietary protein and sugar. PMID- 18396139 TI - Drosophila ALS regulates growth and metabolism through functional interaction with insulin-like peptides. AB - In metazoans, factors of the insulin family control growth, metabolism, longevity, and fertility in response to environmental cues. In Drosophila, a family of seven insulin-like peptides, called Dilps, activate a common insulin receptor. Some Dilp peptides carry both metabolic and growth functions, raising the possibility that various binding partners specify their functions. Here we identify dALS, the fly ortholog of the vertebrate insulin-like growth factor (IGF)-binding protein acid-labile subunit (ALS), as a Dilp partner that forms a circulating trimeric complex with one molecule of Dilp and one molecule of Imp L2, an IgG-family molecule distantly related to mammalian IGF-binding proteins (IGFBPs). We further show that dALS antagonizes Dilp function to control animal growth as well as carbohydrate and fat metabolism. These results lead us to propose an evolutionary perspective in which ALS function appeared prior to the separation between metabolic and growth effects that are associated with vertebrate insulin and IGFs. PMID- 18396140 TI - Transcriptional regulation of adipogenesis by KLF4. AB - While adipogenesis is known to be controlled by a complex network of transcription factors, less is known about the transcriptional cascade that initiates this process. We report here the characterization of Kruppel-like factor 4 (KLF4) as an essential early regulator of adipogenesis. Klf4 is expressed in 3T3-L1 cells within 30 min after exposure to a standard adipogenic cocktail of insulin, glucocorticoids, and IBMX. Knockdown of KLF4 inhibits adipogenesis and downregulates C/EBPbeta levels. KLF4 binds directly to the C/EBPbeta (Cebpb) promoter as shown by chromatin immunoprecipitation and gel shift assays and, together with Krox20, cooperatively transactivates a C/EBPbeta reporter. C/EBPbeta knockdown increases levels of KLF4 and Krox20, suggesting that C/EBPbeta normally suppresses Krox20 and KLF4 expression via a tightly controlled negative feedback loop. KLF4 is specifically induced in response to cAMP, which by itself can partially activate adipogenesis. These data suggest that KLF4 functions as an immediate early regulator of adipogenesis to induce C/EBPbeta. PMID- 18396141 TI - Rapid activation of Akt2 is sufficient to stimulate GLUT4 translocation in 3T3-L1 adipocytes. AB - The serine/threonine kinase Akt2 has been implicated in insulin-regulated glucose uptake into muscle and fat cells by promoting the translocation of glucose transporter 4 (GLUT4) to the cell surface. However, it remains unclear whether activation of Akt2 is sufficient since a role for alternate signaling pathways has been proposed. Here we have engineered 3T3-L1 adipocytes to express a rapidly inducible Akt2 system based on drug-inducible heterodimerization. Addition of the dimerizer rapalog resulted in activation of Akt2 within 5 min, concomitant with phosphorylation of the Akt substrates AS160 and GSK3. Comparison with insulin stimulation revealed that the level of Akt2 activity observed with rapalog was within the physiological range, reducing the likelihood of off-target effects. Transient activation of Akt2 also increased glucose transport and GLUT4 translocation to the plasma membrane. These results show that activation of Akt2 is sufficient to stimulate GLUT4 translocation in 3T3-L1 adipocytes to an extent similar to insulin. PMID- 18396142 TI - A paper-based electroelution system for protein recovery from stained sodium dodecyl sulfate-polyacrylamide gels. AB - Electroelution is a widely used methodology for protein purification. In this study, a practical and low-cost system for protein electroelution from stained polyacrylamide gels was developed. For this, a horizontal protein electroelution cuve was constructed with glass plates, 1.5-ml capacity microcentrifuge tubes, and dialysis membrane. Analyses of the system efficiency showed high protein recovery from nonfixed and fixed sodium dodecyl sulfate (SDS)-polyacrylamide gels. PMID- 18396143 TI - Quartz crystal microbalance analysis of growth kinetics for aggregation intermediates of the amyloid-beta protein. AB - Evidence linking soluble aggregation intermediates of the amyloid-beta protein (A beta), as well as the ongoing growth of A beta aggregates, to physiological responses characteristic of Alzheimer's disease (AD) indicates that a kinetic description A beta aggregation intermediate growth may be fundamental to understanding disease progression. Although the growth of mature A beta fibrils has been investigated using several experimental platforms, the growth of A beta aggregation intermediates has been less thoroughly explored. In the current study, a quartz crystal microbalance (QCM) was employed to analyze the real-time growth of A beta(1-40) aggregation intermediates selectively immobilized on the crystal surface. Immobilization permitted quantitative evaluation of A beta(1-40) aggregation intermediate growth under controlled solution conditions. Elongation of A beta(1-40) aggregation intermediates via monomer addition proceeded in a nonsaturable and reversible fashion. The rate of elongation was observed to vary linearly with both monomer concentration and immobilized aggregate density, to be elevated by increases in solution ionic strength, and to increase as solution pH became more acidic. Elongation was consistent with a first-order kinetic model for the single growth phase observed. These findings extend previous kinetic studies involving the growth of mature A beta fibrils to describe the growth of A beta(1-40) aggregation intermediates via monomer addition. PMID- 18396144 TI - Improved enrichment strategies for phosphorylated peptides on titanium dioxide using methyl esterification and pH gradient elution. AB - Improvements to phosphopeptide enrichment protocols employing titanium dioxide (TiO2) are described and applied to identification of phosphorylation sites on recombinant human cyclin-dependent kinase 2 (CDK2). Titanium dioxide binds phosphopeptides under acidic conditions, and they can be eluted under basic conditions. However, some nonphosphorylated peptides, particularly acidic peptides, bind and elute under these conditions as well. These nonphosphorylated peptides contribute significantly to ion suppression of phosphopeptides and also increase sample complexity. We show here that the conversion of peptide carboxylates to their corresponding methyl esters sharply reduces nonspecific binding, improving the selectivity for phosphopeptides, just as has been reported for immobilized metal affinity chromatography (IMAC) columns. We also present evidence that monophosphorylated peptides can be effectively fractionated from multiply phosphorylated peptides, as well as acidic peptides, via stepwise elution from TiO2 using pH step gradients from pH 8.5 to pH 11.5. These approaches were applied to human CDK2 phosphorylated in vitro by yeast CAK1p in the absence of cyclin. We confirmed phosphorylation at T160, a site previously documented and shown to be necessary for CDK2 activity. However, we also discovered several novel sites of partial phosphorylation at S46, T47, T165, and Y168 when ion-suppressing nonphosphorylated peptides were eliminated using the new protocols. PMID- 18396145 TI - Bridging the regeneration gap: stem cells, biomaterials and clinical translation in bone tissue engineering. AB - Advances in our understanding of skeletal stem cells and their role in bone development and repair, offer the potential to open new frontiers in bone regeneration. Tissue engineering seeks to harness the regenerative capacity innate to bone for the replacement of tissue lost or damaged through a broad range of conditions associated with an increasingly aged population. The strategy entails ex vivo expansion of multipotential populations followed by delivery to the site of damage on dynamically durable-biodegradable three-dimensional structures which provide the requisite extracellular microenvironment for stem cell driven tissue development. This review will examine bone stem cell biology, and current advances in skeletal tissue engineering through the enhancement and marrying of biologically informed and clinically relevant strategies. PMID- 18396146 TI - Interaction of Rab3B with microtubule-binding protein Gas8 in NIH 3T3 cells. AB - Rab3 subfamily small G proteins (Rab3A, Rab3B, Rab3C, and Rab3D) control the regulated exocytosis in neuronal/secretory cells. Rab3B is also detected and upregulated in non-neuronal/non-secretory cells, whereas its function remains elusive. In the present study, we identified growth-arrest-specific gene 8 (Gas8), an evolutionally conserved microtubule-binding protein that is upregulated in growth-arrested NIH 3T3 cells and involved in the dynein motor regulation in flagellar/ciliary axoneme, as a novel Rab3B-binding protein using a yeast two-hybrid system. Rab3B as well as Gas8 was upregulated in growth-arrested NIH 3T3 cells and enriched in testis and lung with well-developed flagella/cilia. Gas8 was specifically interacted with the GTP-bound form of Rab3B and co localized with Rab3B at the Golgi in NIH 3T3 cells. Furthermore, Rab3B was relocated upon expression of the Rab3B-binding domain of Gas8. These results suggest that Gas8 links Rab3B to microtubules in NIH 3T3 cells. PMID- 18396147 TI - CREB and Sp1 regulate the human CD2AP gene promoter activity in renal tubular epithelial cells. AB - The human CD2-associated protein (CD2AP) is involved in several molecular signaling pathways and is an important factor responsible for nephrotic syndrome. Here we report the identification of the transcription start point and promoter region of the human CD2AP gene in renal tubular epithelial cells. With luciferase assays and deletion analysis, we found that the region between -558 and -1bp ahead of the transcription start point is indispensable for the promoter activity of the human CD2AP gene. A CREB site and two Sp1 sites were essential for maintaining the basal transcriptional activity of the human CD2AP promoter. Overexpression of phosphorylated CREB and Sp1 transactivated the human CD2AP promoter, whereas small interfering RNA-mediated blockage of CREB and Sp1 genes expressions inhibited markedly its activity. These findings provide the first analysis of the human CD2AP gene promoter and demonstrate that not only CREB but also Sp1 plays a critical role in regulating basal CD2AP promoter activity in renal tubular epithelial cells. PMID- 18396148 TI - The flavonoid fisetin promotes nerve cell survival from trophic factor withdrawal by enhancement of proteasome activity. AB - To explore the possibility that specific flavonoids can substitute for neurotrophic factors, we examined the ability of the flavonol fisetin and several related flavonoids to support the survival of low density, serum-free cultures of rat cortical neurons. Normally these cells die within 24h in the absence of trophic factors but in the presence of fisetin and several related flavonoids the cells survive and produce long neurites. While the survival-promoting effect of several of the fisetin-related flavonoids was partially dependent on ERK activation, the effect of fisetin was not. Fisetin can enhance glutathione synthesis but the survival-promoting effect of fisetin was also not dependent on glutathione. However, proteasome inhibitors almost completely blocked the ability of fisetin to promote survival. Consistent with this observation, fisetin increased proteasome activity. Together these results demonstrate a new activity for fisetin and tie this activity to its neurotrophic effects. PMID- 18396149 TI - An ancient look at UCP1. AB - Brown adipose tissue serves as a thermogenic organ in placental mammals to defend body temperature in the cold by nonshivering thermogenesis. The thermogenic function of brown adipose tissue is enabled by several specialised features on the organ as well as on the cellular level, including dense sympathetic innervation and vascularisation, high lipolytic capacity and mitochondrial density and the unique expression of uncoupling protein 1 (UCP1). This mitochondrial carrier protein is inserted into the inner mitochondrial membrane and stimulates maximum mitochondrial respiration by dissipating proton-motive force as heat. Studies in knockout mice have clearly demonstrated that UCP1 is essential for nonshivering thermogenesis in brown adipose tissue. For a long time it had been presumed that brown adipose tissue and UCP1 emerged in placental mammals providing them with a unique advantage to survive in the cold. Our subsequent discoveries of UCP1 orthologues in ectotherm vertebrates and marsupials clearly refute this presumption. We can now initiate comparative studies on the structure-function relationships in UCP1 orthologues from different vertebrates to elucidate when during vertebrate evolution UCP1 gained the biochemical properties required for nonshivering thermogenesis. PMID- 18396150 TI - Two domains of the epidermal growth factor receptor are involved in cytoskeletal interactions. AB - Epidermal growth factor receptor can interact directly with F-actin through an actin-binding domain. In the present study, a mutant EGFR, lacking a previously identified actin-binding domain (ABD 1), was still able to bind elements of the cytoskeleton. A second EGFR actin-binding domain (ABD 2) was identified in the region of the receptor that includes Tyr-1148 by a yeast two-hybrid assay. GST fusion proteins comprising ABD 1 or ABD 2 bound actin in vitro and competed for actin-binding with the full-length EGFR. EGFR binding to actin was also studied in intact cells using fluorescence resonance energy transfer (FRET). The localization of the EGFR/actin-binding complex changed after EGF stimulation. Fusion proteins containing mutations in ABD1 or ABD2 did not display a FRET signal. The results lead to the conclusion that the interaction between ABD1 and ABD2 and actin during EGF-induced signal transduction, and thus between EGFR and actin, are important in cell activation. PMID- 18396151 TI - Selective structural change of bulged-out region of double-stranded RNA containing bulged nucleotides by spermidine. AB - Polyamines are essential for cell growth due to effects mainly at the level of translation. These effects likely involve a structural change, induced by polyamines, of the bulged-out region of double-stranded RNA that is different from changes induced by Mg(2+). Structural changes were studied using U6-34, a model RNA of U6 small nuclear RNA containing bulged nucleotides. Binding of NS1-2 peptide derived from the RNA binding site of NS1 protein, to U6-34 was inhibited by spermidine but not by Mg(2+). A selective conformational change of the bases in the bulged-out region of U6-34 induced by spermidine was observed by NMR. The selective effect of spermidine was lost when the bulged-out region of U6-34 was removed in U6-34(Delta5). The binding of NS1-2 peptide to U6-34(Delta5) was inhibited both by spermidine and Mg(2+). The selective structural change of U6-34 by spermidine was confirmed by circular dichroism. PMID- 18396153 TI - Interrogating the role of liposome size in mediating the dynamics of a chromophore in the acyl chain region of a phospholipid bilayer. AB - We have examined the temperature-dependent reorientation dynamics of perylene imbedded in bilayers of 1,2-dimyristoyl-sn-phosphatidylcholine (DMPC), where the bilayers exist in the form of unilamellar vesicles. Previous work using 100-nm diameter DMPC vesicles has shown that the phase transition from the gel phase to the fluid phase can be detected using the reorientation dynamics of perylene. In this work we explore the vesicle size dependence of the perylene reorientation dynamics in DMPC vesicles. The size of the vesicles is determined by extrusion and the reorientation dynamics of perylene are measured as a function of vesicle size between 100-nm and 5-microm diameter. We find that, while the phase transition for DMPC is seen in smaller vesicles, perylene becomes insensitive to the phase transition for vesicles larger than ca. 800-nm diameter. We also find a discontinuous change in perylene reorientation dynamics with increasing vesicle size, and we consider this result in the context of the location of perylene within the bilayer. PMID- 18396152 TI - Structure and dynamics of polyunsaturated hydrocarbon chains in lipid bilayers significance for GPCR function. AB - This review summarizes results of our recent solid-state NMR investigations on polyunsaturated 18:0-22:6n3-PC/PE/PS and 18:0-22:5n6-PC bilayers. Data on structure and dynamics of the polyunsaturated docosahexaenoyl (DHAn3, 22:6n3) and docosapentaenoyl chains (DPAn6, 22:5n6), investigated at physiological conditions, are reported. Lipid-protein interaction was studied on reconstituted bilayers containing the G-protein coupled membrane receptor (GPCR) rhodopsin as well as on rod outer segment (ROS) disk membranes prepared from bovine retinas. Results reveal surprisingly rapid conformational transitions of polyunsaturated chains and existence of weakly specific interactions of DHAn3 with spatially distinct sites on rhodopsin. PMID- 18396154 TI - Association between tumor necrosis factor (TNF)-alpha G-308A gene polymorphism and preeclampsia complicated by severe fetal growth restriction. AB - BACKGROUND: Preeclampsia and HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome are multifactorial disorders with genetic and environmental components. Given that the tumor necrosis factor (TNF)-alpha G-308A single nucleotide polymorphism (SNP) affects TNF-alpha gene transcription and that preeclampsia and HELLP syndrome are characterized by a shift towards a Th1 type maternal immune response with increased TNF-alpha production, the aim of the current study was to investigate whether this SNP is associated with preeclampsia and HELLP syndrome in a Caucasian population from Hungary. Additionally, we aimed to examine whether TNF-alpha G-308A polymorphism can influence the risk for fetal growth restriction in preeclamptic patients, which issue none of the earlier studies dealt with. METHODS: In a case-control study, we analyzed blood samples from 140 preeclamptic patients, 69 patients with HELLP syndrome and 144 normotensive, healthy pregnant women using the polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP) method. We performed also a meta analysis with our results and those of 8 previously published studies. RESULTS: There were no significant differences in the genotype and allele frequencies of the TNF-alpha G-308A polymorphism between preeclamptic patients and normotensive, healthy pregnant women. However, the mutant (TNF2 or A) allele occurred significantly more frequently in preeclamptic patients with IUGR than in those without IUGR (18.5% versus 7.1%, p=0.003). In addition, the frequency of the mutant allele carriers was significantly higher among preeclamptic patients with IUGR compared to those without IUGR (30.6% versus 12.8%, p=0.010). The mutant allele carriers were found to have an increased risk of severe IUGR-complicated preeclampsia, which was independent of maternal age, prepregnancy BMI and primiparity (odds ratio (OR): 2.89, 95% confidence interval (CI): 1.16-7.22, p=0.023; adjusted OR: 2.78, 95% CI: 1.04-7.45, p=0.042). Nevertheless, no significant differences were detected in the genotype and allele frequencies of the TNF-alpha G-308A polymorphism between patients with HELLP syndrome and control subjects. In the meta-analysis, no association was observed between this SNP and preeclampsia (summary OR: 0.956, 95% CI: 0.693-1.319). CONCLUSIONS: Although the meta-analysis demonstrated a lack of an overall association between TNF-alpha G-308A polymorphism and preeclampsia, our results suggest a role of this SNP in the risk of severe IUGR-complicated preeclampsia. However, further studies are required with a larger sample size to confirm our findings. PMID- 18396155 TI - Evaluation of a novel reverse-hybridization StripAssay for typing DNA variants useful in diagnosis of adult-type hypolactasia. AB - BACKGROUND: Adult-type hypolactasia is a genetically determined inability to digest lactose after weaning. Two single-nucleotide polymorphisms (C-13910T, G 22018A) located upstream of the lactase gene (LCT) within the gene MCM6 are associated with the lactase persistence/non-persistence trait in patients of European descent. Therefore, the genotyping of these SNPs has been established as a diagnostic tool for adult-type hypolactasia. We have recently shown that several novel allelic variants located in close proximity to the C-13910T SNP interfere with the diagnostic accuracy of real-time PCR-based genotyping methods. METHODS: We describe here the validation of a comprehensive reverse-hybridization teststrip-based assay for the detection of common and novel LCT SNPs (C-13907G, C 13910T, T-13913C, G-13914A, T-13915G, and G-22018A). This assay is based on multiplex DNA amplification and ready-to-use membrane teststrips containing variant-specific oligonucleotide probes immobilized as an array of parallel lines. RESULTS: We evaluated the novel reverse-hybridization StripAssay on 125 DNA samples in comparison to LightCycler analysis and sequencing. The outcome of StripAssay genotyping was found to be completely concordant with that obtained by sequencing. CONCLUSIONS: The StripAssay represents an accurate and robust screening tool to identify multiple LCT/MCM6 variants in a rapid manner. It overcomes diagnostic pitfalls that were reported and allows the simultaneous genotyping of closely spaced LCT variant sites in a single-step diagnostic approach. PMID- 18396156 TI - Effect of polymorphisms of endothelial nitric oxide synthase on ischemic stroke: a case-control study in a Chinese population. AB - BACKGROUND: Nitric oxide (NO) synthesized by endothelial nitric oxide synthase (eNOS) plays a key role in vascular regulation and atherosclerosis, therefore, eNOS may be a candidate gene for ischemic stroke (IS). However, it is still controversial whether eNOS polymorphisms are a risk factor for IS. METHODS: Three polymorphisms of the eNOS gene (-922A/G, -786T/C, 894G/T) were determined by using TaqMan SNP genotyping assay in 309 Chinese patients with IS and 309 Chinese controls. RESULTS: The frequency of eNOS -922 G allele was significantly higher in the patients than the controls (12.14% vs 8.09%, P=0.018). The distribution of eNOS genotypes differed insignificantly between the 2 groups. The frequency of the eNOS -786 CC genotype was higher in the patients than the controls (OR=3.819, P=0.029). With respect to -922A/G, the AG+GG genotype increased the risk for IS (OR=1.523, P=0.047). After adjustment for confounding factors, the odds ratios of -786 CC and -922 variant genotype (AG+GG) for IS were 4.580 and 1.656, respectively. However, haplotype analysis revealed the frequencies of Hap4 (GCG) and Hap7 (GCT) were significantly higher in the patients than the controls (P=0.035, 0.042). CONCLUSIONS: eNOS -922A/G and -786T/C may affect the susceptibility to IS and certain haplotypes of the eNOS gene may be associated with a higher susceptibility to IS. PMID- 18396157 TI - TAFI antigen level variability in young healthy Asian Indians; first report from Asia. AB - Thrombin Activatable Fibrinolytic Inhibitor (TAFI) is a plasma protein, which inhibits fibrinolysis by removing carboxyterminal lysine residues from partially degraded fibrin thereby decreasing plasminogen binding on its surface. In this study we have investigated the antigenic level variability (Inter and Intraindividual) of Thrombin Activatable Fibrinolysis Inhibitor in 120 healthy Asian Indians since no data on this is available regarding this population. TAFI antigen levels did not show a normal distribution in our population (p<0.001). Median TAFI antigen levels were found to be 11.683 microg/ml. It ranged from 33.9 202.5%of normal pool plasma (3.9-23.5 microg/ml). TAFI antigenic level showed high level of variability in the Indian population (coefficient of variation: 37.4%). TAFI antigenic levels were stable intraindividually (p=0.218). PMID- 18396158 TI - Spectroelectrochemistry of cytochrome b559 in the D1-D2-Cyt b559 complex from spinach. AB - The redox potential of cytochrome b559 (Cyt b559) in the D1-D2-Cyt b559 complex from spinach has been determined to be +90+/-2mV vs. SHE at pH 6.0, by thin-layer cell spectroelectrochemistry for the first time. The redox potential, corresponding uniquely to the so-called "low-potential form", exhibited a sigmoidal pH-dependence from pH 4.0 to 9.0, ranging from +115 to +50mV. An analysis of the pH-dependence based on model equations suggests that two histidine residues coordinating to the heme iron in the protein subunits may exert electrostatic influence on the redox potential of Cyt b559. PMID- 18396159 TI - One SmpB molecule accompanies tmRNA during its passage through the ribosomes. AB - tmRNA and SmpB are the main participants of trans-translation, a process which rescues the ribosome blocked during translation of non-stop mRNA. While a one-to one stoichiometry of tmRNA to the ribosome is generally accepted, the number of SmpB molecules in the complex is still under question. We have isolated tmRNA ribosome complexes blocked at different steps of the tmRNA path through the ribosome and analyzed the stoichiometry of the complexes. Ribosome, tmRNA and SmpB were found in equimolar amount in the tmRNA-ribosome complexes stopped at the position of the 2nd, 4th, 5th or the 11th codons of the coding part of the tmRNA. PMID- 18396160 TI - Intracellular delivery of acetyl-histone peptides inhibits native bromodomain chromatin interactions and impairs mitotic progression. AB - Bromodomains present in Brd4 and other chromatin proteins interact with acetylated histones to regulate transcription and cell growth. To study Brd4 chromatin interactions in vivo, histone H4 tail peptides were fused to a synthetic protein transduction domain (PTD) derived from the human immunodeficiency virus Tat and delivered into cultured cells. Acetyl-H4 peptides, but not unacetylated H4 peptides inhibited real time Brd4-chromatin interactions in living cells as assessed by fluorescence recovery after photobleaching assays. The acetyl-H4 peptides also inhibited an interaction of Brd4 with chromosomes during mitosis and reduced cell growth potential. Together, PTD-based delivery of histone tail peptides offers a novel means to study the mechanism and biological significance of bromodomain-chromatin interactions in vivo. PMID- 18396161 TI - The Lhcb protein and xanthophyll composition of the light harvesting antenna controls the DeltapH-dependency of non-photochemical quenching in Arabidopsis thaliana. AB - Nonphotochemical quenching (NPQ) is the photoprotective dissipation of energy in photosynthetic membranes. The hypothesis that the DeltapH-dependent component of NPQ (qE) component of non-photochemical quenching is controlled allosterically by the xanthophyll cycle has been tested using Arabidopsis mutants with different xanthophyll content and composition of Lhcb proteins. The titration curves of qE against DeltapH were different in chloroplasts containing zeaxanthin or violaxanthin, proving their roles as allosteric activator and inhibitor, respectively. The curves differed in mutants deficient in lutein and specific Lhcb proteins. The results show that qE is determined by xanthophyll occupancy and the structural interactions within the antenna that govern allostericity. PMID- 18396162 TI - Cytochrome c is released from coupled mitochondria of yeast en route to acetic acid-induced programmed cell death and can work as an electron donor and a ROS scavenger. AB - To gain insight into the processes by which acetic acid-induced programmed cell death (AA-PCD) takes place in yeast, we investigated both cytochrome c release from yeast mitochondria and mitochondrial coupling over the time course of AA PCD. We show that the majority of cytochrome c release occurs early in AA-PCD from intact coupled mitochondria which undergo only gradual impairment. The released cytochrome c can be reduced both by ascorbate and by superoxide anion and in turn be oxidized via cytochrome c oxidase, thus working both as a ROS scavenger and a respiratory substrate. Late in AA-PCD, the released cytochrome c is degraded. PMID- 18396163 TI - Role of the repressor JDP2 in the amino acid-regulated transcription of CHOP. AB - The transcriptional activation of CHOP (C/EBP-homologous protein) by amino acid deprivation involves ATF2 and ATF4 binding at the amino acid response element within the promoter. In this report, we investigate the role of JDP2 (Jun Dimerization Protein 2) in the amino acid control of CHOP transcription following amino acid starvation. Our results show that JDP2 binds to the CHOP AARE in unstimulated cells and that its binding decreases following amino acid starvation. We demonstrate that JDP2 acts as a repressor and suggest that it could be functionally associated with HDAC3 to inhibit CHOP transcription. PMID- 18396164 TI - Possible role of a taurine transporter in the deep-sea mussel Bathymodiolus septemdierum in adaptation to hydrothermal vents. AB - Various invertebrates inhabiting hydrothermal vents possess sulfur-oxidizing bacteria in their tissues; however, the mechanisms by which toxic sulfides are delivered to these endosymbionts remain unknown. Recently, detoxification of sulfides using thiotaurine, a sulfur-containing amino acid, has been suggested. In this study, we propose the involvement of a taurine transporter in sulfide detoxification in the deep-sea mussel Bathymodiolus septemdierum by demonstrating: (i) the abundance of its mRNA in the gill; (ii) its activity under a wide range of salinities; (iii) its low Michaelis constant value in taurine transportation; and (iv) its affinity for thiotaurine and the thiotaurine precursor, hypotaurine. PMID- 18396165 TI - Hydrogen-deuterium exchange strategy for delineation of contact sites in protein complexes. AB - We use NMR spectra to determine protein-protein contact sites by observing differences in amide proton hydrogen-deuterium exchange in the complex compared to the free protein in solution. Aprotic organic solvents are used to preserve H/D labeling patterns that would be scrambled in water solutions. The binding site between the mammalian co-chaperone Aha1 with the middle domain of the chaperone Hsp90 obtained by our H/D exchange method corresponds well with that in the X-ray crystal structure of the homologous complex from yeast, even to the observation of a secondary binding site. This method can potentially provide data for complexes with unknown structure and for large or dynamic complexes inaccessible via NMR and X-ray methods. PMID- 18396166 TI - Lil3 assembles as chlorophyll-binding protein complex during deetiolation. AB - Dark-grown angiosperm seedlings are etiolated and devoid of chlorophyll. Deetiolation is triggered by light leading to chlorophyll dependent accumulation of the photosynthetic machinery. The transfer of chlorophyll to the chlorophyll binding proteins is still unclear. We demonstrate here that upon illumination of dark-grown barley seedlings, two new pigment-binding protein complexes are de novo accumulated. Pigments bound to both complexes are identified as chlorophyll a and protochlorophyll a. By auto-fluorescence tracking and mass spectrometry, we show that exclusively Lil3 is the pigment-binding complex subunit in both complexes. PMID- 18396167 TI - Delineating the susceptibility of botulinum neurotoxins to denaturation through thermal effects. AB - Botulinum neurotoxins (BoNT) are the etiological agents responsible for botulism and are acknowledged terrorist threat agents. Passive immunotherapy may provide one countermeasure. Importantly, in the virtually unlimited repertoire of antibody specificities, enzyme linked immunosorbent assays (ELISA) has become an indispensable method for antibody selection. We report that of the BoNTs, BoNT/E is highly susceptible to polystyrene induced denaturation. To further dissect this result and the potential susceptibility of other BoNTs to denaturation we selected a thermal platform, which could be readily quantified using surface plasmon resonance (SPR), a primary rat spinal cord cell-based assay and an animal lethality model. PMID- 18396168 TI - Lamellipodia and filopodia in metastasis and invasion. AB - Advances in our understanding of the mechanisms of lamellipodia and filopodia assembly have led to a better concept of how cells move, including how the actin cytoskeleton might be important for the motility of metastatic cancer cells. The cytoskeleton is a potentially interesting target for prevention of invasion and metastasis. As key proteins are uncovered which regulate the assembly of actin based structures, these need to be considered in light of whether they represent potential invasion and metastasis proteins. PMID- 18396170 TI - Molybdate transport through the plant sulfate transporter SHST1. AB - Molybdenum is an essential micronutrient required by plants. The mechanism of molybdenum uptake in plants is poorly understood, however, evidence has suggested that sulfate transporters may be involved. The sulfate transporter from Stylosanthes hamata, SHST1, restored growth of the sulfate transport yeast mutant, YSD1, on media containing low amounts of molybdate. Kinetic analysis using 99MoO4(2-) demonstrated that SHST1 enhanced the uptake of molybdate into yeast cells at nM concentrations. Uptake was not inhibited by sulfate, but sulfate transport via SHST1 was reduced with molybdate. These results are the first measurement of molybdate transport by a characterised plant sulfate transport protein. PMID- 18396169 TI - Indole and other aromatic compounds activate the yeast TRPY1 channel. AB - The yeast TRPY1 (Yvc1p) channel is activated by membrane stretch to release vacuolar Ca2+ into the cytoplasm upon osmotic upshock. Exogenously added indole greatly enhances the upshock-induced Ca2+ release in vivo. Indole also reversibly activates the channels under patch clamp. A minimum of 10(-6)M Ca2+ is needed for membrane stretch force to open TPRY1, but indole activation appears to be Ca2+ independent. A deletion of 30 residues at the predicted cytoplasmic domain, 570 600Delta, renders TRPY1 insensitive to stretch force upto 10(-3)M Ca2+. Nonetheless, indole readily activates this mutant channel. Several other aromatic compounds, e.g. the antimicrobial parabens, also activate TRPY1. These compounds likely alter the innate forces in the lipid bilayer received by the channel. PMID- 18396171 TI - Neuronal nitric oxide synthase: prototype for pulsed enzymology. AB - The established paradigm in understanding and describing enzyme activity uses formalisms based on steady-state assumptions, including Michaelis-Menten and King Altman approaches. These are appropriate for enzymes operating under steady-state conditions. Signal generating enzymes transfer information, rather than material. Because the information capacity of a signal channel depends on frequency, steady state descriptions may not be appropriate. Recently, Stuehr and coworkers described a novel product inhibition mechanism for NO synthases. Simulations presented here suggest that at physiological temperatures neuronal nitric oxide synthase produces sharp pulses of NO, consistent with its signaling function. These temporal pulses greatly restrict the effective spatial range of NO signaling. PMID- 18396172 TI - Dietary carbohydrate restriction induces a unique metabolic state positively affecting atherogenic dyslipidemia, fatty acid partitioning, and metabolic syndrome. AB - Abnormal fatty acid metabolism and dyslipidemia play an intimate role in the pathogenesis of metabolic syndrome and cardiovascular diseases. The availability of glucose and insulin predominate as upstream regulatory elements that operate through a collection of transcription factors to partition lipids toward anabolic pathways. The unraveling of the details of these cellular events has proceeded rapidly, but their physiologic relevance to lifestyle modification has been largely ignored. Here we highlight the role of dietary input, specifically carbohydrate intake, in the mechanism of metabolic regulation germane to metabolic syndrome. The key principle is that carbohydrate, directly or indirectly through the effect of insulin, controls the disposition of excess dietary nutrients. Dietary carbohydrate modulates lipolysis, lipoprotein assembly and processing and affects the relation between dietary intake of saturated fat intake and circulating levels. Several of these processes are the subject of intense investigation at the cellular level. We see the need to integrate these cellular mechanisms with results from low-carbohydrate diet trials that have shown reduced cardiovascular risk through improvement in hepatic, intravascular, and peripheral processing of lipoproteins, alterations in fatty acid composition, and reductions in other cardiovascular risk factors, notably inflammation. From the current state of the literature, however, low-carbohydrate diets are grounded in basic metabolic principles and the data suggest that some form of carbohydrate restriction is a candidate to be the preferred dietary strategy for cardiovascular health beyond weight regulation. PMID- 18396174 TI - Developments in instrumentation. Letter from the editors. PMID- 18396173 TI - Effect of lipid peroxidation products on the activity of human retinol dehydrogenase 12 (RDH12) and retinoid metabolism. AB - Mutations in human Retinol Dehydrogenase 12 (RDH12) are known to cause photoreceptor cell death but the physiological function of RDH12 in photoreceptors remains poorly understood. In vitro, RDH12 recognizes both retinoids and medium-chain aldehydes as substrates. Our previous study suggested that RDH12 protects cells against toxic levels of retinaldehyde and retinoic acid [S.A. Lee, O.V. Belyaeva, I.K. Popov, N.Y. Kedishvili, Overproduction of bioactive retinoic acid in cells expressing disease-associated mutants of retinol dehydrogenase 12, J. Biol. Chem. 282 (2007) 35621-35628]. Here, we investigated whether RDH12 can also protect cells against highly reactive medium-chain aldehydes. Analysis of cell survival demonstrated that RDH12 was protective against nonanal but not against 4-hydroxynonenal. At high concentrations, nonanal inhibited the activity of RDH12 towards retinaldehyde, suggesting that nonanal was metabolized by RDH12. 4-Hydroxynonenal did not inhibit the RDH12 retinaldehyde reductase activity, but it strongly inhibited the activities of lecithin:retinol acyl transferase and aldehyde dehydrogenase, resulting in decreased levels of retinyl esters and retinoic acid and accumulation of unesterified retinol. Thus, the results of this study showed that RDH12 is more effective in protection against retinaldehyde than against medium-chain aldehydes, and that medium-chain aldehydes, especially 4-hydroxynonenal, severely disrupt cellular retinoid homeostasis. Together, these findings provide a new insight into the effects of lipid peroxidation products and the impact of oxidative stress on retinoid metabolism. PMID- 18396175 TI - Developments in instrumentation. Guest editorial. PMID- 18396176 TI - Positron emission tomography/computed tomography. AB - Accurate anatomical localization of functional abnormalities obtained with the use of positron emission tomography (PET) is known to be problematic. Although tracers such as (18)F-fluorodeoxyglucose ((18)F-FDG) visualize certain normal anatomical structures, the spatial resolution is generally inadequate for accurate anatomic localization of pathology. Combining PET with a high-resolution anatomical imaging modality such as computed tomography (CT) can resolve the localization issue as long as the images from the two modalities are accurately coregistered. However, software-based registration techniques have difficulty accounting for differences in patient positioning and involuntary movement of internal organs, often necessitating labor-intensive nonlinear mapping that may not converge to a satisfactory result. Acquiring both CT and PET images in the same scanner obviates the need for software registration and routinely provides accurately aligned images of anatomy and function in a single scan. A CT scanner positioned in line with a PET scanner and with a common patient couch and operating console has provided a practical solution to anatomical and functional image registration. Axial translation of the couch between the 2 modalities enables both CT and PET data to be acquired during a single imaging session. In addition, the CT images can be used to generate essentially noiseless attenuation correction factors for the PET emission data. By minimizing patient movement between the CT and PET scans and accounting for the axial separation of the two modalities, accurately registered anatomical and functional images can be obtained. Since the introduction of the first PET/CT prototype more than 6 years ago, numerous patients with cancer have been scanned on commercial PET/CT devices worldwide. The commercial designs feature multidetector spiral CT and high performance PET components. Experience has demonstrated an increased level of accuracy and confidence in the interpretation of the combined study as compared with studies acquired separately, particularly in distinguishing pathology from normal, physiologic tracer uptake and precisely localizing abnormal foci. Combined PET/CT scanners represent an important evolution in technology that has helped to bring molecular imaging to the forefront in cancer diagnosis, staging and therapy monitoring. PMID- 18396177 TI - Respiratory motion in positron emission tomography/computed tomography: a review. AB - The development of positron emission tomography/computed tomography (PET/CT) scanners has allowed not only straightforward but also synergistic fusion of anatomical and functional information. Combined PET/CT imaging yields an increased sensitivity and specificity beyond that which either of the 2 modalities possesses separately and therefore provides improved diagnostic accuracy. Because attenuation correction in PET is performed with the use of CT images, with CT used in the localization of disease, accurate spatial registration of PET and CT image sets is required. Correcting for the spatial mismatch caused by respiratory motion represents a particular challenge for the requisite registration accuracy as a result of differences in temporal resolution between the 2 modalities. This review provides a brief summary of the materials, methods, and results involved in multiple investigations of the correction for respiratory motion in PET/CT imaging of the thorax, with the goal of improving image quality and quantitation. Although some schemes use respiratory-phase data selection to exclude motion artifacts, others have adopted sophisticated software techniques. The various image artifacts associated with breathing motion are also described. PMID- 18396179 TI - Positron emission tomography/magnetic resonance imaging: the next generation of multimodality imaging? AB - Multimodal imaging is now well-established in routine clinical practice. Especially in the field of nuclear medicine, new positron emission tomography (PET) installations comprise almost exclusively combined PET/computed tomography (CT) scanners rather than PET-only systems. However, PET/CT has certain notable shortcomings, including the inability to perform simultaneous data acquisition and the significant radiation dose to the patient contributed by CT. Magnetic resonance imaging (MRI) offers, compared with CT, better contrast among soft tissues as well as functional-imaging capabilities. Therefore, the combination of PET with MRI provides many advantages that go far beyond simply combining functional PET information with structural MRI information. Many technical challenges, including possible interference between these modalities, have to be solved when combining PET and MRI, and various approaches have been adapted to resolving these issues. Here, we present an overview of current working prototypes of combined PET/MRI scanners from different groups. In addition, besides PET/MRI images of mice, the first such images of a rat acquired with the first commercial clinical PET/MRI scanner, are presented. The combination of PET and MRI is a promising tool in preclinical research and will certainly progress to clinical application. PMID- 18396180 TI - Small-animal preclinical nuclear medicine instrumentation and methodology. AB - Molecular medicine enhances the clinician's ability to accurately diagnose and treat disease, and many technological advances in diverse fields have made the translation of molecular medicine to the clinic possible. Nuclear medicine encompasses 2 technologies--single-photon emission computed tomography (SPECT) and positron emission tomography (PET)--that have driven the field of molecular medicine forward. SPECT and PET, inherently molecular imaging techniques, have been at the forefront of molecular medicine for several decades. These modalities exploit the radioactive decay of nuclides with specific decay properties that make them useful for in vivo imaging. As recently as the mid-1990s, SPECT and PET were mostly restricted to use in the clinical setting because their relatively coarse spatial resolution limited their usefulness in studying animal (especially rodent) models of human disease. About a decade ago, several groups began making significant strides in improving resolution to the point that small-animal SPECT and PET as a molecular imaging technique was useful in the study of rodent disease models. The advances in these 2 techniques progressed as the result of improvements in instrumentation and data reconstruction software. Here, we review the impact of small-animal imaging and, specifically, nuclear medicine imaging techniques on the understanding of the biological basis of disease and the expectation that these advances will be translated to clinical medicine. PMID- 18396178 TI - Technological development and advances in single-photon emission computed tomography/computed tomography. AB - Single-photon emission computed tomography/computed tomography (SPECT/CT) has emerged during the past decade as a means of correlating anatomical information from CT with functional information from SPECT. The integration of SPECT and CT in a single imaging device facilitates anatomical localization of the radiopharmaceutical to differentiate physiological uptake from that associated with disease and patient-specific attenuation correction to improve the visual quality and quantitative accuracy of the SPECT image. The first clinically available SPECT/CT systems performed emission-transmission imaging using a dual headed SPECT camera and a low-power x-ray CT subsystem. Newer SPECT/CT systems are available with high-power CT subsystems suitable for detailed anatomical diagnosis, including CT coronary angiography and coronary calcification that can be correlated with myocardial perfusion measurements. The high-performance CT capabilities also offer the potential to improve compensation of partial volume errors for more accurate quantitation of radionuclide measurement of myocardial blood flow and other physiological processes and for radiation dosimetry for radionuclide therapy. In addition, new SPECT technologies are being developed that significantly improve the detection efficiency and spatial resolution for radionuclide imaging of small organs including the heart, brain, and breast, and therefore may provide new capabilities for SPECT/CT imaging in these important clinical applications. PMID- 18396182 TI - Sex differences in depression symptoms in treatment-seeking adults: confirmatory analyses from the Sequenced Treatment Alternatives to Relieve Depression study. AB - BACKGROUND: Although epidemiologic research consistently reports greater prevalence of major depressive disorder in women, small sample sizes in many studies do not allow for full elaboration of illness characteristics. This article examines sex differences in terms of illness attributes in a cohort of 2541 outpatients from across the United States who enrolled in the Sequenced Treatment Alternatives to Relieve Depression study. METHODS: Confirmatory analyses were performed in 2541 outpatients comparing men and women with regard to sociodemographic features, comorbid Axis I and Axis III conditions, and illness characteristics. Results were compared with those of our previous report on the initial population of the first 1500 individuals enrolled in Sequenced Treatment Alternatives to Relieve Depression study. RESULTS: In both samples, nearly two thirds of the sample (62.5%) were women. Women had greater symptom severity, but men had more episodes of major depression, despite no difference in the length of illness. No differences in age of onset emerged. As in the first cohort, women showed greater rates of an anxiety disorder, bulimia, and somatoform disorder, as well as more past suicide attempts, whereas men showed more alcohol and substance abuse. Women reported more appetite, weight, hypersomnia, interpersonal sensitivity, gastrointestinal and pain complaints, and less suicidal ideation. Irritability was equally common in men and women. CONCLUSION: This large analysis confirmed most of the clinical features and comorbidities found to be more prevalent in the first cohort of women. In addition, this analysis corroborated previous research suggesting higher rates of atypical and anxious depression in women but refuted the notion of an "irritable depression" found in men. The report confirmed the 1.7:1 ratio for depression seen across sexes in the National Comorbidity Survey. PMID- 18396183 TI - A family history study of binge-eating disorder. AB - Family studies of anorexia nervosa and bulimia nervosa have yielded important information about the etiologies of these eating disorders. By contrast, little is known about familial factors of etiologic importance for binge-eating disorder (BED). The purpose of the current family history study was to assess the prevalence of comorbid psychopathology in a non-treatment seeking female sample of 31 probands with BED, 32 control probands without BED, and their 283 first degree relatives. In-person semistructured clinical interviews were conducted with the probands, who also served as informants for all of their first-degree relatives. Significantly higher lifetime rates of major depressive disorder, dysthymic disorder, and social phobia were found among women with BED compared with control women. Significantly higher lifetime rates of bipolar (I or II) disorder, any depressive disorder, nearly all anxiety disorders, anorexia nervosa, and BED were reported among the first-degree relatives of women with BED compared with the first-degree relatives of control women. Furthermore, female relatives of women with BED were reported to have higher rates of substance use disorders and dysthymic disorder compared with female relatives of control women without BED. Nearly all disorders that were elevated in relatives of women with BED followed a pattern of independent transmission from BED. The primary exception was substance use disorder among female relatives, whose transmission pattern was consistent with that of a shared etiology with BED. Thus, BED and substance use disorder may share a common mechanism of familial transmission among women. PMID- 18396184 TI - Incidence and diagnostic stability of ICD-10 acute and transient psychotic disorders. AB - OBJECTIVE: The 10th Revision of the International Classification of Diseases (ICD 10) introduced a new diagnostic category, F23 acute and transient psychotic disorders (ATPD) to embrace clinical concepts such as the French bouffee delirante, Kleist and Leonhard's cycloid psychoses, and the Scandinavian reactive and schizophreniform psychoses. The relative rarity of these disorders and insufficient follow-up studies with adequate numbers of patients makes ATPD classification as uncertain as their validity. The aim of this study was to evaluate incidence and validity of ATPD in terms of diagnostic stability. METHOD: A 6-year analysis of readmission patterns of all subjects listed in the Danish psychiatric central register as having been first-ever admitted to hospital or treated in outpatient services with a diagnosis of ATPD from January 1 to December 31, 1996, was conducted. RESULTS: The incidence of ATPD was 9.6 per 100 000 population, with a higher rate of females than males (9.8 vs 9.4). Incidence rates by age group were higher for males than for females, with a marked reversal of this pattern above 50 years. This contrasted with incidence of schizophrenia that was almost twice as high in males as in females, particularly in the 20-29 year age group. Of 416 cases with a first-admission diagnosis of ATPD, an increasing number tended to change on subsequent admissions, nearly half to another F2 category schizophrenia and related disorders. The overall stability rate reached only 39%. CONCLUSIONS: Although demographic differences from schizophrenia are topics that deserve further research, poor diagnostic stability argues against attempts to separate ATPD from borderland disorders. PMID- 18396181 TI - Antisocial behavioral syndromes and body mass index among adults in the United States: results from the National Epidemiologic Survey on Alcohol and Related Conditions. AB - OBJECTIVE: To describe associations of antisocial behavioral syndromes, including DSM-IV antisocial personality disorder (ASPD) and conduct disorder without progression to ASPD ("CD only"), and syndromal antisocial behavior in adulthood without CD before age 15 (AABS, not a codable DSM-IV disorder), with body mass index (BMI) status in the general US adult population. METHODS: This report is based on the 2001-2002 National Epidemiologic Survey on Alcohol and Related Conditions (n = 43093; response rate, 81%). Respondents were classified according to whether they met criteria for ASPD, AABS, "CD only," or no antisocial syndrome, and on current BMI status based on self-reported height and weight. Associations of antisocial syndromes with BMI status were examined using multinomial logistic regression. RESULTS: Among men, antisociality was not associated with BMI. Among women, ASPD was significantly associated with overweight and extreme obesity; AABS was associated with obesity and extreme obesity; and "CD only" was significantly associated with overweight, obesity, and extreme obesity. CONCLUSIONS: Assessment of antisocial features appears warranted in overweight, obese, and extremely obese women, and assessment of BMI status appears indicated in antisocial women. Prevention and treatment guidelines for overweight and obesity may need revision to address comorbid antisociality, and interventions targeting antisociality may need to include attention to weight concerns. PMID- 18396185 TI - Clinical profile of "schizo-obsessive" disorder: a comparative study. AB - Obsessive-compulsive disorder (OCD) is a common comorbid condition in schizophrenia. The clinical implications of this comorbidity are uncertain. There is some evidence to suggest that schizophrenia with OCD (schizo-obsessive) has a poor prognosis, but the effect of OCD on schizophrenia symptom profile is unclear. Therefore, we studied the clinical profile of schizophrenic patients with and without comorbid OCD. Fifty consecutive patients who met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for both schizophrenia and OCD and an equal number of schizophrenic patients without OCD were recruited over a period of 12 months from the clinical services of the National Institute of Mental Health and Neuro Sciences, Bangalore, India. Their clinical profile was systematically assessed and compared to detect significant differences, if any. Schizo-obsessive patients were more likely to have paranoid symptoms and first-rank symptoms of schizophrenia. They had lower anergia, higher depression scores, more comorbid personality disorders, and somewhat lesser disability. Significant correlations were observed between the Yale-Brown Obsessive-Compulsive Scale scores (including the Yale-Brown Obsessive-Compulsive Scale insight score) and schizophrenia symptom dimension scores. Our findings are concordant with the hypothesis that "schizo-obsessive" schizophrenia may be a distinct subtype with unique clinical characteristics, supporting the need for further research in this area. PMID- 18396186 TI - The course of dissociation in the general population: a 3-year follow-up study. AB - OBJECTIVE: There have been no previous general population studies on the stability of dissociative symptoms. The aim of this study was to examine the course of and the changes in dissociative symptoms and factors associated with these changes during a 3-year follow-up of a Finnish general population sample. METHODS: The general population sample included a cohort of 1497 subjects. Dissociative symptoms were assessed with the Dissociative Experiences Scale (DES) and the DES-taxon (DES-T). Depressive symptoms and suicidal ideation were measured with the Beck Depression Inventory (BDI). The sample was categorized into low dissociators with DES scores less than 20 and high dissociators with DES scores of 20 or more. RESULTS: At baseline, 98 subjects were high dissociators. On follow-up, 28 of them were still high dissociators, whereas among 70 subjects, the DES score declined below the cutoff score. During the follow-up period, 28 of 1399 subjects became new high dissociators, and constantly low dissociators consisted of 1371 of 1399 subjects. Dissociative taxon membership was detected in 39 subjects either at baseline or at follow-up, but only 4 of them met the criteria at both assessments. Stable high dissociation was associated with an increase in the BDI score on follow-up, baseline suicidal ideation, a younger age, a reduced working ability, and smoking. Risk factors for becoming a new high dissociator were an increase in the BDI score, a younger age at baseline, and a reduced working ability. Among the baseline high dissociators, recovery from high dissociation was associated with a decline in the BDI score at follow-up and with no suicidal thoughts, older age, and a good working ability at baseline. CONCLUSIONS: Only a small proportion of the general population had constantly high levels of dissociative symptoms. The stability of dissociative taxon membership was weaker than the stability of the continuous variables of dissociation. The dissociative experiences had a tendency to change, and these changes were associated with changes in the BDI scores. Further studies are needed to reveal the factors associated with the changes in dissociative symptoms. PMID- 18396187 TI - A 6-month study of postpartum depression and related factors in Athens Greece. AB - INTRODUCTION: Postpartum depression (PPD) affects women in various sociocultural environments around the world during a sensitive period of their lives. The purpose of this study was to investigate the prevalence and time course of PPD in a Greek urban environment as well as possible relations of PPD with certain clinical and sociodemographic factors. METHOD: The study was performed on a sample of 402 women that were recruited from a university obstetric clinic in Athens, Greece, during the first 24 hours after delivery. The women completed the Edinburgh Postnatal Depression Scale through telephone interviews. The telephone interviews were conducted the first week as well as the first, third, and sixth month after delivery. The first day after delivery, all women completed the Montgomery-Asberg Depression Rating Scale, the List of Threatening Experience, the State-Trait Anxiety Inventory, the Whitley Index, the Schalling-Sifneos Personality Scale, and the Maudsley Obsessive-Compulsive Inventory. In addition, the Blues Questionnaire was administered the first 3 days and the seventh day after delivery. Other clinical and sociodemographic data were obtained through questionnaires and personal interviews. RESULTS: A cutoff point of 12 in the Edinburgh Postnatal Depression Scale was used to define PPD. Eighty (19.8%) of the women in the sample experienced PPD during the first 6 months after delivery. The development of PPD was related significantly to the following factors: stressful events during pregnancy (P = .01), maternity blues on the seventh day after delivery (P = .01), obsessive preoccupation with cleaning (P = .04), and judgment that the baby is crying excessively at the first month interview (P = .02). CONCLUSION: The women's emotional condition before and after delivery, obsessionality, and difficulties in regulating the infant's emotions appear to contribute to the development of PPD during the first 6 months after delivery. PMID- 18396188 TI - Agoraphobia between panic and phobias: clinical epidemiology from the Sesto Fiorentino Study. AB - In the last few decades, there has been a long debate on the existence of agoraphobia (AG) without a history of panic attacks (PAs). In the present study, the problem of the relationships between AG and PAs is addressed trough a reevaluation of the cases who had been diagnosed with AG in the community survey of Sesto Fiorentino. Forty-one of the 75 subjects who met the criterion of AG in the Sesto Fiorentino Study were reinterviewed by experienced clinical psychiatrists. The Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) and the Composite International Diagnostic Interview were used to make the diagnoses. The Mobility Inventory for Agoraphobia (MIA) and a specific adjunctive question, "why do/did you avoid?", were used to compare AG subjects with or without PD. Of the 41 subjects with a lifetime history of AG, 12 cases had original diagnosis of AG without PAs and the remaining 29 had PD with AG. After the reassessment, in 10 cases, the criteria for the diagnosis of AG without PAs were confirmed, totaling a lifetime prevalence of 0.4% (confidence interval, 0.2-0.8). Agoraphobia subjects with and without PAs were comparable as regard to sex, age, age of onset, duration of illness, family history for anxiety or mood disorders, MIA scores, number, and type of situations avoided. Thus, AG seems to exist also in absence of a history of PAs, and the one-way relationship between the occurrence of PAs and a following development of AG, postulated by DSM-IV, should be reconsidered for the future classifications. PMID- 18396189 TI - Sense of coherence among delusional patients: prediction of remission and risk of relapse. AB - Delusional diseases are characterized by a remitting/relapsing course, which, in current studies, is not contextualized with experience and well-being. This study applied the salutogenic model in order to predict the course of delusional disease, using the "sense of coherence" (SOC) concept, which reflects patients' experiences of their disorder. In a prospective study, 48 delusional schizophrenic inpatients were interviewed, using self-report scales for delusions, SOC, depression and expressed emotion (EE), at 3 time points over a 1 year period, with an additional 6-month follow-up based on treatment staff reports and medical files. Sense of coherence was found to be strongly correlated with delusional symptoms over the study period. Higher SOC scores at the "acute delusional state" predicted better prognosis with regard to delusions. However, SOC was found to be lower during remission, suggesting decreased well-being during remission. Sense of coherence was found to be a stronger predictor of 1 year prognosis for delusions when EE or depression were high at the acute state. A complex relationship was found between delusions, SOC, depression, and EE, suggesting possible use of the SOC scale as a prognostic tool. In addition, the results stress the necessity of an integrative biopsychosocial treatment approach, which would include interventions aimed at enhancing elements of SOC, particularly during periods of remission. To the best of our knowledge, this is the first longitudinal study of SOC in delusional patients moving through remissions and relapses. PMID- 18396190 TI - Posttraumatic stress disorder; combat exposure; and nicotine dependence, alcohol dependence, and major depression in male twins. AB - Combat exposure is associated with increased risk of psychiatric and substance use disorders in veterans. However, it is not known whether combat exposure independently increases risk for these disorders or whether this association is accounted for by genetic vulnerability common to posttraumatic stress disorder (PTSD). This article tests competing explanations for the association of combat exposure and PTSD with nicotine dependence (ND), alcohol dependence (AD), and major depression (MD). Data were obtained from 6099 members of the Vietnam Era Twin Registry, a national registry of male-male twin pairs who served in the military during the Vietnam era. Twin models were fit to estimate the genetic and environmental variance common and specific to Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition, lifetime diagnoses of PTSD, combat trauma, and 3 comorbid conditions: ND, AD, and MD. Variance specific to ND, AD, and MD was due to genetic factors (48%, 36%, and 12%, respectively) and unique environmental factors (36%, 42%, and 58%, respectively). After accounting for variance common to PTSD, no residual genetic and environmental variance overlapped between combat and ND, combat and AD, and combat and MD. Combat exposure is not independently associated with lifetime ND, AD, and MD. The association of combat exposure with these 3 disorders is due to genetic and unique environmental contributions in common with PTSD. These findings suggest comorbid PTSD may represent a genetically mediated vulnerability to psychopathology after trauma. PMID- 18396191 TI - Clinical implications of insight assessment in obsessive-compulsive disorder. AB - Poor insight has been reported in 15% to 36% of patients with obsessive compulsive disorder (OCD), but little is known about its clinical correlations. This study examines insight among patients with OCD using a standardized instrument, the Brown Assessment of Beliefs Scale, and analyzes its relationship with clinical factors. Insight was assessed in 132 patients with OCD, before and after pharmacologic treatment, using the Brown Assessment of Beliefs Scale. Differences between patients with good and poor insight on sociodemographic variables, OCD severity, comorbidity, and treatment response were studied. Stability of insight after pharmacologic treatment was also examined. Thirty-nine patients (29.5%) exhibited poor insight. They showed more depressive symptoms (P = .001) and personality disorders (P = .001), especially the schizotypal form, than did good insight subjects, but there were no significant differences in treatment response. Insight significantly improves after treatment (P < .001). Our results suggest that insight in OCD varies widely and constitutes a dynamic phenomenon that can improve after treatment and is influenced by clinical conditions such as affective status or personality. PMID- 18396192 TI - Reliability and validity of the Korean version of the Davidson Trauma Scale. AB - The Davidson Trauma Scale (DTS) is a validated, 17-item, brief global assessment scale for posttraumatic stress disorder (PTSD). The purposes of this study were to develop a Korean version of the DTS (DTS-K) while maintaining its basic structure and to evaluate its reliability and validity for the Korean population. Participants of this study included 93 patients with PTSD (PTSD group), 73 patients with nonpsychotic mood or other anxiety disorders (psychiatric control group), and 88 healthy controls (normal control group). Subjects completed psychometric assessments, including the DTS-K and the Korean version of the Clinician-Administered PTSD Scale and the State Trait Anxiety Inventory. The DTS K showed good internal consistency (Cronbach alpha = .97) and test-retest reliability (r = .93). The DTS-K showed a significantly positive correlation with Clinician-Administered PTSD Scale (r = .94). The highest diagnostic efficiency of DTS-K was at a total score of 47, with sensitivity and specificity of 0.87 and 0.84, respectively. Our findings suggest that the DTS-K is composed of good psychometric properties and is a valid and reliable tool for assessing the frequency and severity of PTSD symptoms regardless of ethnicity. PMID- 18396193 TI - The erythropoiesis-stimulating agents: reimbursement challenges and more. PMID- 18396194 TI - Introduction: role of surgery in the diagnosis and management of metastatic cancer. PMID- 18396195 TI - Current neurosurgical management of brain metastases. AB - Brain metastases occur in a third of patients diagnosed with cancer; without any intervention, the prognosis is quite poor with a median survival of 1 month. Because of the constraints of the blood-brain barrier, chemotherapy is not effective and treatment options include surgery, whole brain radiation, or stereotactic radiation. This chapter is devoted to a review of the current management options for treatment of brain metastases. PMID- 18396196 TI - The role of surgery in the management of metastatic spinal tumors. AB - The role of surgery in the treatment of metastatic spinal tumors causing epidural compression traditionally consisted of posterior decompression. This procedure plus radiotherapy, however, could not be demonstrated to provide any benefit over radiotherapy alone, and surgery fell into disfavor in managing metastatic vertebral tumors. The advent of newer, more sophisticated approaches, along with improved spinal instrumentation and reconstruction techniques, which allowed direct decompression of neural elements and resection of the tumor, have revived the use of surgery in these tumors. These modern spinal surgery techniques, in combination with radiotherapy, have yielded significantly superior functional outcomes and prolonged survival in symptomatic metastatic epidural compression when compared to radiotherapy alone. Management of spinal metastases is evolving, and a multitude of factors determine the indication for and the technique and goals of surgical intervention. Between 1993 and 2005, 21.1% of patients with metastatic spinal tumors evaluated at The University of Texas M.D. Anderson Cancer Center were treated surgically by the Department of Neurosurgery. The most common spinal metastasis operated upon was metastatic kidney cancer (31.5%), even though kidney cancer was only the third most common primary tumor (after lung and breast cancers) giving rise to vertebral metastases observed during the same time period at this institution. This highlights the importance of the histology of the primary cancer (among other factors) in determining the indication for surgical intervention. PMID- 18396197 TI - Current surgical management of bone metastases in the extremities and pelvis. AB - Surgical management of metastases to the extremities and pelvis has benefited from advances in the technology of internal fixation, as well as the increased availability of options for large endoprostheses. Contoured periarticular plates and the screws that attach rigidly to the plates have made fixation into weakened bone more reliable and easier to provide. For massive bone loss, modular endoprostheses are now widely available. These options supplemented with bone cement (polymethylmethacrylate) give patients the ability to have most bone defects reinforced or replaced such that the patient can begin using the affected limb almost immediately. PMID- 18396198 TI - The role of thoracoscopy in the management of cancer patients. AB - Thoracoscopy or video-assisted thoracic surgery (VATS) has become an important tool in the management of thoracic diseases. Most thoracic procedures are first attempted using a VATS technique. Thoracoscopy has an important role in managing malignancies that present with symptoms or lesions in the chest. Although the primary role is diagnostic, in the case of non-thoracic malignancies, VATS also has an important palliative treatment role, as in dealing with pleural and pericardial effusions and metastatic deposits. PMID- 18396199 TI - Surgical treatment of metastatic disease to the lung. AB - The presence of distant metastases usually implies disease not amenable to cure through surgical resection. In such cases, chemotherapy is the mainstay of treatment, with surgery or radiation reserved for palliative measures. However, metastases limited to the lung may be resected with resultant prolonged patient survival compared to unresectable, widely disseminated metastases. Isolated pulmonary metastases should therefore not be considered untreatable. In this review, we discuss the pathophysiology of pulmonary metastases. We outline prognostic factors associated with metastases, and propose criteria to help select patients for metastasectomy. Surgical approaches, including various open techniques and video-assisted thoracoscopy, are covered. Surgical issues, including the need for unilateral versus bilateral exploration, the extent of resection to achieve cure, the need for lymph node dissection, and the benefit of repeat operations, are discussed. Finally, we review some of the more common tumors that metastasize to the lungs, and the role of metastasectomy in their treatment. Resection of pulmonary metastases confers a survival benefit to a select group of patients so long as the primary tumor is controlled, metastases are limited to the lungs, the patient can tolerate the operation from a cardiopulmonary standpoint, and the metastases are completely resected. PMID- 18396200 TI - Diagnosis and treatment of metastatic disease to the liver. AB - A number of cancers present with synchronous or metachronous hepatic metastases. Historically, many of these patients were considered unresectable and were treated with either systemic chemotherapy or supportive care. Today, a variety of options exist for the management of hepatic metastases. Newer agents for systemic therapy continue to be introduced and are providing improved progression-free and overall survival and increased resectability of liver metastases. However, complete surgical resection of isolated hepatic metastases remains the optimal management for these patients. Surgical interventions can be offered to patients with hepatic-only metastases. Hepatic artery chemotherapy represents an adjunct for those patients undergoing resection and can improve survival. This benefit may be even more pronounced when combined with systemic chemotherapy. Newer generation biologic agents can improve results. New therapeutic modalities to treat lesions that are unresectable include ablative techniques such as radiofrequency ablation (RFA) and cryoablation. This article will examine modalities of diagnosis of hepatic metastases and highlight the data regarding hepatic resection for metastases of several types of primary cancers, the rationale for, and efficacy of, hepatic arterial chemotherapy, in both the postoperative adjuvant setting and in unresectable liver disease, and review the current literature for ablative techniques in the treatment of liver metastases. PMID- 18396201 TI - Metastatic disease to the pancreas and spleen. AB - Isolated metastases to the pancreas and spleen are a rare occurrence. When they are diagnosed, pancreatic metastases are most often from renal cell carcinoma, lung cancer, and breast cancer. The most common source of splenic metastases is gynecological in origin; the overwhelming majority is ovarian. If extensive staging studies reveal these metastases to be isolated, then curative resection may be warranted. This review will demonstrate that long-term survival may be achieved in patients with isolated metastases and a prolonged disease-free interval. PMID- 18396202 TI - Surgical management of metastatic disease to the adrenal gland. AB - Metastatic disease to the adrenal glands can occur in a wide array of malignancies. With the increased use of abdominal imaging, these lesions are diagnosed with more frequency. Diagnostic and laboratory evaluation is essential for the differentiation of benign lesions from primary malignant adrenal tumors or extra-adrenal metastasis. Computed tomography (CT) and magnetic resonance imaging (MRI) characteristics, as well as the adjunctive use of immunocytochemical techniques on biopsy specimens, can allow accurate identification of metastatic lesions. Surgical management of metastatic lesions is appropriate in selected patients, primarily when representing the solitary site of metastatic disease. The surgical approach, while debatable, can de done either through open surgery or laparoscopically. Either approach appears comparable in terms of oncologic efficacy in the carefully selected patient, although laparoscopic adrenalectomy is associated with decreased pain and improved convalescence. The surgeon's skill in laparoscopic technique, appropriate patient selection, and the ability to adhere to oncologic principles, including complete excision without tumor spillage, are of utmost importance when deciding the appropriate surgical intervention. PMID- 18396203 TI - Current surgical management of intestinal metastases. AB - Malignant tumors of the small bowel account for less than 10% of all gastrointestinal neoplasms. The majority of these tumors are secondary neoplasms metastatic from either an intra- or extra-abdominal site. Symptoms related to these tumors include bleeding, obstruction, and perforation, although the great majority of these metastases remain asymptomatic and are only discovered at autopsy. Outcomes remain poor and most interventions are palliative. We review the current surgical recommendations for management of this challenging clinical scenario. PMID- 18396204 TI - Diagnosis and management of peritoneal carcinomatosis arising from adenocarcinoma of the colon and rectum. AB - Peritoneal carcinomatosis (the implantation of tumor cells throughout the peritoneal cavity) occurs in as many as 50% of patients with colorectal cancer and in up to 25% is the only site of recurrence. There is increasing evidence that aggressive debulking of peritoneal only disease followed by intraperitoneal chemotherapy can have significant impact on the patient's quality and sometimes quantity of life. This chapter will provide an overview of the biologic basis of pathogenesis and the treatment of peritoneal metastases from colorectal cancer. PMID- 18396205 TI - Positive association of anticytoskeletal endothelial cell antibodies and cardiac allograft rejection. AB - Using an indirect immunofluorescence method on human umbilical vein endothelial cells (HUVEC), we investigated the presence of antiendothelial cell antibodies (AECA) in 136 pre- and posttransplant serum samples sequentially collected from 31 patients during the first year after cardiac transplantation. A healthy control group was also included (n = 87). Colocalization studies demonstrated a positive staining pattern of different cytoskeletal components (cytoskeletal antiendothelial cell antibodies, CSK-AECA) including antivimentin, antiactin, antitubulin, and anticytokeratin among heart transplanted patients. Frequency of CSK-AECA in the control group and at day 0 in the transplant group was 18.3 and 22.5%, respectively (p = NS). A progressive increase in the frequency of CSK-AECA was observed after cardiac transplantation: 13.3% at day 15; 22.2% at day 30; 53.8% at day 90, and 58% at day 360. Interestingly, rejection episodes within the first year after transplantation occurred in 83.3% of CSK-AECA-positive and in 30.7% of CSK-AECA-negative patients (p = 0.0045). The presence of antibodies was detected prior to the rejection event and was associated with a poor clinical outcome: rejection episodes occurred at a mean of 36.14 +/- 17 days after transplantation in patients with preexisting AECA and 174.25 +/- 51.9 days after de novo antibody appearance in patients with no antibodies at day 0 (p = 0.029). In conclusion, a progressive increase in the frequency of CSK-AECA was observed following cardiac transplantation; the presence of these antibodies is strongly associated and precedes the rejection episodes. Thus, CSK-AECA could be a good marker for acute graft rejection. PMID- 18396206 TI - 17 Beta-estradiol (E2) plus tumor necrosis factor-alpha induces a distorted maturation of human monocyte-derived dendritic cells and promotes their capacity to initiate T-helper 2 responses. AB - There is growing evidence that 17 beta-estradiol (E2) modulates immune function. Recent studies indicated that certain effects of E2 on in vivo immune function are not a result of a direct action on T cells, but rather an indirect action on antigen-presenting cells. This study demonstrates that the pregnancy-associated doses of E2 plus tumor necrosis factor-alpha (TauNuF alpha) induce distorted maturation of human dendritic cells (DCs) that result in an increased capacity to induce T helper (Th) 2 responses. E2 did not affect the expression of human leukocyte antigen class II and costimulatory molecules by DCs, but elicited the ability of DC to produce CC chemokine ligand 1, which can attract CCR8-expressing Th2 cells and regulatory T cells. In addition, E2/TNF alpha-matured DCs increased the production of IL-10 relative to the IL-12p70 on CD40 ligation, thereby inducing naive T-cell differentiation into a Th2. Moreover, the increased concentration of E2 in the route of maturation did indeed further enhance Th2 deviation. The dominant Th2 deviation was induced at a high E2 concentration typical during pregnancy. These findings demonstrate that the high physiological levels of E2 may be an important endogenous component for regulating the DC function and skewing the immune response toward Th2. PMID- 18396208 TI - Suppressive dendritic cells as a tool for controlling allograft rejection in organ transplantation: promises and difficulties. AB - The most important antigen-presenting cells are dendritic cells (DCs), which play a central role in the initiation of immunity and tolerance. Their immunoregulatory properties offer the potential of donor-specific control of graft rejection after organ transplantation. It has not been clarified which DC subpopulations mediate tolerance, and the use of natural DCs for therapeutic applications is therefore problematic. Suppressive DCs can be generated in vitro by treating the cells with biologic, pharmacologic, or genetic agents. Here we discuss approaches for generating inhibitory DCs and present DC-based animal models for control of allograft rejection. A prerequisite of suppressive DCs for therapeutic application in clinical transplantation is a reproducible method for their generation as well as the induction of irreversible suppressive function. Based on lessons learned from the use of DCs as tools in clinical vaccine trials in cancer, we discuss the unknown aspects and risks of DC therapy in transplantation. PMID- 18396207 TI - Immunoglobulin allotypes influence IgG antibody responses to hepatitis C virus envelope proteins E1 and E2. AB - Immunoglobulin (Ig) GM and KM allotypes-genetic markers of gamma and kappa chains, respectively-are associated with the outcome of hepatitis C virus (HCV) infection, but the underlying mechanisms are not well understood. We hypothesized that GM and KM allotypes could contribute to the outcome of HCV infection by influencing the levels of IgG antibodies to the HCV glycoproteins E1E2. We serologically allotyped 100 African American individuals with persistent HCV infection for GM and KM markers and measured anti-E1E2 antibodies. Subjects with the GM 1,17 5,13 phenotype had significantly higher levels of anti-E1E2 antibodies than subjects who lacked this phenotype (p = 0.008). Likewise, subjects with the KM 1-carrying phenotypes had higher levels of anti-E1E2 antibodies than subjects who lacked these phenotypes (p = 0.041). Median titers were fourfold higher in persons expressing both GM 1,17 5,13 and KM 1-carrying phenotypes compared with those who lacked these phenotypes (p = 0.011). Interactive effects of these GM-KM phenotypes were previously found to be highly significantly associated with spontaneous HCV clearance. Results presented here show that Ig allotypes contribute to the interindividual differences in humoral immunity to the HCV epitopes, a finding that may provide a mechanistic explanation for their involvement in the outcome of HCV infection. PMID- 18396209 TI - Limited allelic diversity of stimulatory two-domain killer cell immunoglobulin like receptors. AB - Genomic sequencing was used to characterize most of the coding regions of the five two-domain stimulatory killer cell immunoglobulin-like receptor (KIR) loci from 80 unrelated, primarily Caucasian, individuals. Specific loci were present in from 26% (KIR2DS3) to 98% (KIR2DS4) of individuals. The number of known alleles present varied from one (KIR2DS1, KIR2DS5) to five (KIR2DS4). The frequencies of loci and alleles were similar to observations made in populations of European and Asian ethnicities. New alleles were found at 2DS1 (*00202, *00302, *005, *006, *007) and 2DS4 (*008) loci. PMID- 18396210 TI - Potential influence of interleukin-1 haplotype IL-1 beta-511*T-IL-1RN*1 in conferring low risk to middle third location of esophageal cancer: a case-control study. AB - Interleukin (IL)-1 gene polymorphisms affect several inflammatory diseases, including cancer. Therefore, we studied genetic association of biallelic ( 511C>T) polymorphism of IL-1 beta and 86-bp VNTR polymorphism of IL-1RN in 159 patients with esophageal cancer (EC) and 194 age- and gender-matched healthy controls. Genetic analysis for IL-1 polymorphisms was performed by polymerase chain reaction-restriction fragment length polymorphism. The frequencies of IL-1 beta (-511C>T) and IL-1RN (variable number tandem repeat) genotypes, alleles, and haplotypes did not differ significantly between patients and controls. However, IL-1 beta -511TT genotype and T1+ haplotype combination illustrated low risk for disease at the middle third location of the tumor (odds ratio [OR] = 0.27; 95% confidence interval [CI] = 0.11-0.62; p = 0.002; OR = 0.462; 95% CI = 0.253 0.845, p = 0.01). In conclusion, subjects with IL-1 beta -511TT genotype or IL-1 beta*T-IL-1RN*1 (T1) haplotype had lower risk for middle third tumor location of EC in a northern Indian population. PMID- 18396211 TI - Interleukin-1 gene complex single nucleotide polymorphisms in systemic sclerosis: a further step ahead. AB - Gene-gene and gene-environment interactions are difficult to detect by traditional parametric computational approaches. Novel nonparametric and model free strategies, such as the multifactor dimensionality reduction (MDR) algorithm, are thus emerging as practical and feasible methods of analysis to model high-order epistatic interactions, integrating and complementing traditional logistic approaches. With traditional methods of analysis we showed that the interleukin-1 beta (IL-1 beta) C+3962T single nucleotide polymorphism (SNP), along with the Sc70 antibody and the diffuse cutaneous subset of systemic sclerosis, are important risk factors for the development of a severe ventilatory restriction in patients with systemic sclerosis (SSc); however the interactions among these and other genetic and environmental attributes were difficult to model. On the contrary, the MDR analysis detected significant two- or three-way interactions in the presence of nonlinearity. The best model identified by the multifactor dimensionality reduction algorithm included the antibody subset, the IL-1 beta C-511T and the interferon-gamma AUTR5644T SNPs, with a testing accuracy of 85% (p < 0.001) and a cross-validation consistency of 10/10. This model outperformed any one- to-three-way model constructed by considering the three factors with main independent effects identified by traditional computational approaches. Epistatic interactions among IL-1 gene complex SNPs and clinical or environmental factors are more important than the singe attributes in the development of severe ventilatory restriction in SSc patients. PMID- 18396212 TI - Association studies of CTLA-4, CD28, and ICOS gene polymorphisms with B-cell chronic lymphocytic leukemia in the Polish population. AB - Abnormal expression of the costimulatory molecules cytotoxic T-lymphocyte antigen 4 (CTLA-4), CD28, and inducible co-stimulator (ICOS) leads to disturbances of immune response and an increased risk of cancer. An extended study was undertaken to evaluate the association among the polymorphisms CTLA-4c.49A>G, CTLA 4g.319C>T, CTLA-4g.*642AT(8_33), CD28c.17+3T>C, and ICOSc.1554+4GT(8_15) and susceptibility to B-cell chronic lymphocytic leukemia (B-CLL) in the Polish population. The study revealed increased frequency of the CTLA-4g.319C>T [T] allele and the CTLA-4g.319C>T [T] phenotype in B-CLL patients compared with healthy controls (p = 0.003, odds ratio [OR] = 1.73; and p = 0.009, OR = 1.74, respectively). The presence of the CD28c.17+3T>C [C] allele and the CD28c.17+3T>C [C] phenotype increased the OR of B-CLL to 1.59 (p = 0.007) and 1.74 (p = 0.007), respectively. Either CTLA-4g.319C>T or CD28c.17+3T>C was associated with time to Rai stage progression. The distributions of the alleles and genotypes of the ICOS gene significantly differed between patients and controls (p = 0.0009 and p = 0.006, respectively). Individuals possessing short alleles were 2.02 times more prone to B-CLL than others (p = 0.001), whereas carriers of long alleles were protected from B-CLL (p = 0.02, OR = 0.62). The haplotype association study and multivariate analysis confirmed the association of CTLA-4g.319C>T and ICOSc.1554+4GT(8_15) gene polymorphisms with B-CLL. The polymorphic sites CTLA 4c.49A>G and CTLA-4g.*642AT(8_33) did not correlate with B-CLL. Our results are the first in the literature to report that gene polymorphism of the costimulatory molecules CTLA-4, CD28, and ICOS contributes to susceptibility to B-CLL. PMID- 18396213 TI - Paroxysmal nocturnal hemoglobinuria: significant association with specific HLA-A, -B, -C, and -DR alleles in an Italian population. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by the expansion of a PIG-A mutated hematopoietic stem cell. An immune-mediated origin has been suggested for this disease. Because HLA genes represent a susceptibility factor for autoimmunity, we investigated HLA genotype in 42 Italian PNH patients compared with 301 control subjects of the same ethnic origin. A significantly increased frequency of the HLA class I alleles A*0201 (p < 0.05), B*1402 (p < 0.001), and Cw*0802 (p < 0.005), and of the HLA class II DRB1*1501 (p < 0.01) with the linked DQB1*0602 (p 65 years and almost 10% of people aged >80 years. OBJECTIVE: The goal of this study was to identify risk factors for bleeding during warfarin treatment of nonvalvular atrial fibrillation (NNVAF) in older patients (those aged >or=75 years) compared with younger patients (those aged <75 years) in clinical practice. METHODS: All patients with NVAF newly started on warfarin at an anticoagulation clinic in a large university hospital were included in this prospective observational study. Patient details were recorded at their first visit; details of any bleeding events were recorded via telephone interview every 4 to 6 weeks for a minimum of 10 months. Patients were divided into 2 groups (ie, those >or=75 years old and those <75 years old). Logistic regression analysis was used to identify risk factors for bleeding. RESULTS: A total of 402 patients were included in the study. Group I comprised 203 patients <75 years old (mean [SD] age, 64.33 [8.33] years) and group II comprised 199 patients >or=75 years old (mean [SD] age, 80.44 [3.99] years). Follow-up ranged from 1 to 31 months (mean [SD], 19 [8.11] months). For major bleeding, number of medications was a significant risk factor in older patients (odds ratio [OR], 3.0; 95% CI, 1.2-7.8 [P = 0.02 ]) and range of the international normalized ratio (INR) was a significant risk factor in both groups. For every unit increase in the range of INR, the odds of major bleeding increased by 0.6 (OR, 1.6; 95% CI, 1.2-2.4 [P = 0.03 ]) in younger patients and by 0.4 (OR, 1.4; 95% CI, 1.07-1.99 [P = 0.04 ])in older patients. For minor bleeding, history of hypertension was the only significant risk factor in older patients (OR, 3.3; 95% CI, 1.3-8.1 [P = 0.01 ]), while history of ischemic heart disease was the only risk factor in younger patients (OR, 1.9; 95% CI, 1.1-5.4 [P = 0.04 ]). CONCLUSIONS: Bleeding pattern was similar in both age groups regarding severity, onset, anatomic site of bleeding, and INR values during the bleeding event. Risk factors for episodes of major bleeding, which are more of a clinical concern, are potentially modifiable. They include quality of anticoagulation control in both groups and number of medications in the older age group. PMID- 18396244 TI - Antidepressant use in nonmajor depression: secondary analysis of a program to encourage active, rewarding lives for seniors (PEARLS), a randomized controlled trial in older adults from 2000 to 2003. AB - BACKGROUND: It is estimated that major depressive disorder affects 0.9% of community-dwelling older adults in the United States. However, as many as 18% of older US adults reportedly suffer from depressive symptoms that do not necessarily fit the criteria for major depressive disorder (eg, dysthmia, minor depression). OBJECTIVES: The goals of this study were to describe patterns of antidepressant medication use in older adults with dysthymia or minor depression and to examine factors associated with the use of antidepressants at baseline. METHODS: This was a secondary analysis using cross-sectional data collected during a randomized controlled trial conducted from 2000 through 2003. It involved community senior service agencies and in-home visits in Seattle, Washington. Adults aged >or=60 years who had minor depression or dysthymia and were receiving services through community senior service agencies or living in senior public housing were included. Study participants were classified as users or nonusers of antidepressants. Prescription medication use in the past 2 weeks was assessed at baseline and 6 and 12 months. Medication name, dose, and directions were recorded from the medication label. Logistic regression was used to examine variables associated with baseline antidepressant use. RESULTS: A total of 138 patients (mean age, 73.00 years) were included; the majority of study participants were female (779.00%). Overall, 42.33% were nonwhite (34.3% black, 4.4% Asian, 1.5% American Indian/Alaskan Native, 0.7% Hispanic, and 1.5% other). At baseline, 36.2% of study participants (n = 50) were using antidepressants. Selective serotonin reuptake inhibitors were the most common class of antidepressants, used by 62.00%, 70.22%, and 71.11% of antidepressant users at baseline, 6, and 12 months, respectively. However, nortriptyline was the most common antidepressant at baseline, taken by 20.00% of antidepressant users. Use of other prescription medications was associated with antidepressant use at baseline. CONCLUSIONS: We found antidepressant use to be low in these relatively poor, community-dwelling, ethnically diverse older adults with dysthymia and minor depression in 2000 through 2003, with 36.22% of participants using antidepressants at baseline. Antidepressant users were more likely to be taking other prescription medications than nonusers. PMID- 18396245 TI - Inappropriate medication use in the elderly: results from a quality improvement project in 99 primary care practices. AB - BACKGROUND: The use of potentially inappropriate medications (PIMs) in the elderly population is common. Interventions to decrease PIM use in primary care settings are needed. OBJECTIVE: This study was designed to assess the time trends in use of always inappropriate and rarely appropriate medications in primary care patients aged >or=65 years during a quality improvement project. METHODS: A 4 year, prospective demonstration project was delivered to 99 primary care practices that use a common electronic medical record and are members of the Practice Partner Research Network. Each participating practice received quarterly performance reports on the use of always inappropriate and rarely appropriate medications in the elderly. Optional interventions included biannual on-site visits and annual network meetings for performance review, academic detailing, and quality improvement planning. General linear mixed regression models were used to analyze the change in prescribing rates over time. RESULTS: Across 42 months of project exposure, 124,802 active patients (61% women, 39% men) aged >or=65 years were included in the analyses. Among the 33 practices that participated in all 42 months of the intervention, the proportion of patients with a prescription for an always inappropriate medication decreased from 0.41% to 0.33%, and the proportion of patients with a prescription for a rarely appropriate medication decreased from 1.48% to 1.30%. Across all 99 practices, the adjusted absolute annual declines for the comprehensive categories of always inappropriate medications (00.018%, P = 0.03) and rarely appropriate medications (0.113%, P = 0.001) were statistically significant. Propoxyphene was the only individual medication that decreased significantly in use over time (baseline proportion, 0.72%; adjusted absolute annual decline, 0.072% [P = 0.001]). CONCLUSIONS: Always inappropriate and rarely appropriate medication use decreased over time in this practice-based research network study. Additional studies of robust interventions for improving medication use in the elderly are warranted. PMID- 18396247 TI - Intracerebral hemorrhage secondary to a warfarin-metronidazole interaction. AB - BACKGROUND: It has been >25 years since the interaction between warfarin and metronidazole was last reported in the literature. The current case report represents the first documentation of this interaction associated with intracerebral hemorrhage. CASE SUMMARY: We present a case of a 78-year-old white woman started on metronidazole (250 mg every 8 hours for 5 days) and levofloxacin (500 mg QD for 6 days) for an upper respiratory tract infection after visiting a walk-in clinic. The patient did not notify any of the health care professionals involved that she was on concomitant warfarin therapy, which had been stable over the last 3 months. Her warfarin dose was 7 mg daily, and her most recent international normalized ratio (INR) reading was 2.5. Nine days after her clinic visit, the patient was admitted to the hospital for a profuse nosebleed with an INR of 8.0 and was found to have an intraparenchymal hemorrhage of the left occipital lobe. The Naranjo adverse drug reaction probability scale indicated that the association with metronidazole was probable and the association with levofloxacin was possible (scores of 7 and 4, respectively). After a 1-week hospital stay, she was discharged. CONCLUSIONS: This adverse event is highly suggestive of a drug interaction caused primarily by metronidazole, which produces an increase in S-warfarin concentrations. Treatment provided by health care providers who were not familiar with the patient and the use of a different pharmacy (where the pharmacist was unaware of her current medications) likely contributed to the event. PMID- 18396249 TI - Editorial: holism--remembering what it is to be human! PMID- 18396246 TI - Behavioral changes with paranoia in an elderly woman taking atorvastatin. AB - BACKGROUND: There have been a number of published reports of central nervous system (CNS) adverse effects with statins. CASE SUMMARY: A 79-year-old woman developed paranoia, anxiety, and behavioral changes approximately 2.5 weeks after starting atorvastatin 10 mg/d. The patient had no other medication changes at this time. After 2 months of therapy, the patient discontinued atorvastatin, and her symptoms fully resolved after 4 days. CONCLUSIONS: This is the first case report, to our knowledge, describing paranoia as one of the symptoms associated with statin therapy. Our report suggests an adverse reaction due to the initiation of atorvastatin via the temporal relationship between the start of atorvastatin and symptom onset, as well as termination of therapy and subsequent symptom disappearance. Use of the Naranjo adverse drug reaction probability scale to assess causality revealed a "probable" association (score, 5) for this adverse event. This report emphasizes the possibility of paranoia as a CNS adverse effect due to statin therapy. Statins are frequently used in older populations and should therefore be considered when such CNS adverse effects occur during therapy. PMID- 18396250 TI - Antibacterial activity of honey against community-associated methicillin resistant Staphylococcus aureus (CA-MRSA). AB - Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has now been described globally, as a clinically significant pathogen, particularly associated with skin and soft tissue infections, including abscesses, cellulitis and furunculosis. The recent emergence of CA-MRSA combined with its predominant presentation associated with skin and soft tissue infection, the previous literature indicating honey as an effective treatment of healthcare-associated HA MRSA-related wound infection, as well as honey's ease of topical application, make the current study timely and of interest to healthcare practitioners involved with wound management. Although previous studies have examined the antimicrobial activity of honey against HA-MRSA, such data are limited regarding the activity of honey against this emerging type of MRSA. CA-MRSA (n=6 isolates), was examined for its susceptibility to natural honey (n=3 honey produced from bees in Northern Ireland and one commercial French honey). Results demonstrated that all honey was able to reduce the cultural count of all CA-MRSA from approximately 10(6) colony-forming units (cfus) (mean = 6.46 log10 cfu/g) to none detectable within 24h of co-culture of separate CA-MRSA organisms individually with all four-honey types examined. Subsequent non-selective enrichment of honey demonstrated that inoculated honey remained positive for CA-MRSA until 72h postinoculation, after which point no culturable organisms could be detected. This study demonstrated that, in vitro, these natural products had an antimicrobial activity against the CA-MRSA organisms tested. Further studies are now required to demonstrate if this antimicrobial activity has any clinical application. PMID- 18396251 TI - Electrodermal activity as a possible physiological marker for acupuncture. AB - Change in amplitude of skin potential is one of the physiological indicators of electrodermal activity (EDA) and has been associated with the onset of a variety of sensory, cognitive and emotional stimuli. This study investigated the EDA physiological response to manual acupuncture. A group of 60 healthy female volunteers were recruited into the study of which 30 were randomly assigned to a control group (no acupuncture) and 30 to the experimental group (received acupuncture). The experimental group attended three sessions in which acupuncture intervention consisting of unilateral manual stimulation of acupuncture points LU7 and KD6. Results showed that the insertion, stimulation and withdrawal of the acupuncture needles were associated with significant changes in EDA. The insertion of the needle was associated with the highest change in skin potential while the three consecutive manipulations of the needles showed a decline in EDA amplitude, thought to be consistent with physiological habituation. Anxiety level and previous familiarity with acupuncture did not influence outcomes. It is postulated that the change in EDA during needle insertion is non-specific to the type and the function of acupuncture points; however, EDA response may be associated with the precise location of the acupuncture point. If further studies confirm such findings, then EDA may become a valuable physiological marker for the acupuncture phenomenon. PMID- 18396252 TI - Prayer in Iranian cancer patients undergoing chemotherapy. AB - Chronic illnesses such as cancer are very stressful events and prayer is a valuable coping strategy for Muslim cancer patients. This study was conducted to assess the impact of prayer in Iranian cancer patients undergoing chemotherapy. A descriptive cross sectional survey was conducted on 360 cancer patients over 20 years of age who were able to read and write. Data collection employed Meraviglia's prayer questionnaire. Mean scores of prayer activity, prayer experience and attitude toward prayer were, respectively, (94.5+/-12.98), (51.2+/ 7.49) and (38.2+/-4.84). Mean of total scores was (184.05+/-21.67). Significant relationships were observed between age, marital status and educational level with prayer activity, prayer experience and attitude toward prayer (P< or = 0.001). Also the relationship between sex and prayer activity was significant (P< or = 0.01). CONCLUSION: Individual characteristics had an important effect on prayer. It is suggested that health care providers can augment holistic care to these patients by encouraging prayer. PMID- 18396253 TI - Survey of parents on the use of naturopathic medicine in children- characteristics and reasons. AB - This cross-sectional study assessed the sociodemographic characteristics of families whose children used naturopathic medicine, the reasons for use, and whether naturopathic and conventional medicine were combined in treating children's conditions. Data were collected in British Columbia, Alberta and Ontario, Canada. Ninety-eight completed questionnaires were analyzed. The results showed that participants (parents) were most likely to be females, university educated, had household income >$60,000, and also saw a naturopathic doctor for themselves. The most common conditions for which children saw a naturopathic doctor included allergies, digestive problems and skin problems, and the most important reasons for use included using all possible options and having a more holistic approach to care. Most parents reported combining naturopathic and conventional care for their children. Study limitations, in particular, selection bias, were discussed. PMID- 18396254 TI - Yoga during pregnancy: effects on maternal comfort, labor pain and birth outcomes. AB - This study examined the effects of a yoga program during pregnancy, on maternal comfort, labor pain, and birth outcomes. A randomized trial was conducted using 74-primigravid Thai women who were equally divided into two groups (experimental and control). The yoga program involved six, 1-h sessions at prescribed weeks of gestation. A variety of instruments were used to assess maternal comfort, labor pain and birth outcomes. The experimental group was found to have higher levels of maternal comfort during labor and 2h post-labor, and experienced less subject evaluated labor pain than the control group. In each group, pain increased and maternal comfort decreased as labor progressed. No differences were found, between the groups, regarding pethidine usage, labor augmentation or newborn Apgar scores at 1 and 5 min. The experimental group was found to have a shorter duration of the first stage of labor, as well as the total time of labor. PMID- 18396255 TI - Influence of race and ethnicity on alternative medicine as a self-treatment preference for common medical conditions in a population of multi-ethnic urban elderly. AB - The use of complementary and alternative medicine (CAM) is prevalent among elderly individuals. While race and ethnicity may influence the choice of CAM, it is uncertain how this influence affects an individual's choice of CAM or conventional medicine. Furthermore, it is unclear whether this choice of CAM or conventional medicine might vary for different medical conditions. A survey of CAM use was performed on a convenience sample of 338 multi-ethnic urban elderly subjects who attended clinic at two large university-affiliated hospitals over 2 years. The survey asked about individual CAM therapies used, and whether subjects would prefer conventional (prescription or over-the-counter) or CAM (herbal or other) treatment for three different medical conditions: colds, insomnia, and back pain. Hispanic ethnicity and female gender were the best predictors of CAM use. Blacks were more likely than whites to utilize CAM. Hispanics were more likely to choose herbal medications to self-treat colds and insomnia than whites or blacks, or low-back pain than whites. More Hispanics chose herbal medications to treat insomnia than over-the-counter or prescription medications. PMID- 18396256 TI - Reflexology treatment for patients with lower limb amputations and phantom limb pain--an exploratory pilot study. AB - OBJECTIVES: The objectives of the study were to evaluate the possibility of reflexology being used as a non-invasive form of phantom limb pain relief and of empowering patients to maintain any positive results with self-treatment. SETTING: Prosthetic Services Centre, Herbert Street, Wolverhampton, West Midlands, England. DESIGN: A same-subject, experimental pilot study, recording the intensity of phantom limb pain in weekly pain diaries over a 30-week period, which was divided into five phases: phase 1 gave a baseline of pain, whilst phase 3 was a resting phase. Phases 2, 4 and 5 provided the reflexology interventions. SAMPLE: Ten participants with unilateral lower limb amputations and phantom limb pain were selected from the database at the Prosthetic Centre. REFLEXOLOGY INTERVENTIONS: In phase 2, six weekly reflexology treatments were given, which consisted of: full foot reflexology to the remaining foot and full hand reflexology to the hand of the amputated side of the body. In phase 4, six weekly hand reflexology teaching sessions were carried out; patients copied on their own hands what the therapist did on hers. A hand reflexology booklet gave the sequence of the treatment and was used as a reference. In phase 5, the patients self-treated for 6 weeks at home, using the reference material. RESULTS: Over the 30-week period, there was an improvement in the perception of the presence and the intensity of the phantom limb pain, with a corresponding improvement in the duration of the pain and the affect on the person's lifestyle. The improvement was maintained when the clients self-treated. FOLLOW-UP QUESTIONNAIRE: A follow up questionnaire was carried out in 2007--12 months after the project had ended- to elicit whether the patients had suffered from phantom pain over the previous 12 months, whether they still had relief from phantom limb pain and whether they still self-treated. CONCLUSIONS: The project indicated that reflexology treatment, teaching and self-treatment were effective in eradicating or reducing the intensity and duration of phantom limb pain, in this group of clients. The follow-up questionnaire revealed that there was a maintained improvement in the intensity of phantom limb pain the patients experienced and that the majority still self-treated. PMID- 18396257 TI - Adverse effects of phytoestrogens on reproductive health: a report of three cases. AB - BACKGROUND: Phytoestrogens have been thought to have favorable effects on women's health and perhaps in offsetting cancers. The possible adverse effects of phytoestrogens have not been evaluated. CASES: Abnormal uterine bleeding with endometrial pathology in three women was found to be related to a high intake of soy products. The first woman had postmenopausal bleeding with uterine polyp, proliferative endometrium and a growing leiomyoma. The second woman presented with severe dysmenorrhea, abnormal uterine bleeding, endometriosis and uterine leiomyoma not responding to treatment. The third woman with severe dysmenorrhea, abnormal uterine bleeding, endometriosis and uterine leiomyomata presented with secondary infertility. All three women improved after withdrawal of soy from their diet. CONCLUSION: Additional information on phytoestrogens is necessary to ascertain their safety before they can be routinely used as supplements. PMID- 18396258 TI - Chair massage for patients and carers: a pilot service in an outpatient setting of a cancer care hospital. AB - PURPOSE: To gather patient and carer evaluations of a 20 min chair massage treatment provided one afternoon a week in an outpatient waiting area. METHOD: Information gathered over a year included documented evaluation of chair massage, pre- and post-treatment well-being scores (visual analogue scale). RESULTS: Both patients (n=224) and carers (n=185) positively evaluated the treatment. Key benefits reported included: relaxation, comfort, time out/treat, distraction, and relief of anxiety. There were significant changes in self-reported well-being score (p=<0.001), but no significant changes between scores for males and females. The changes in well-being scores on occasions (n=3) did not match the positive feedback. CONCLUSIONS: The findings suggest that the service was appreciated by patients and carers. The project was continued for a further year with internal funding. Further research is warranted to ascertain the added and longer-term value of this intervention. PMID- 18396259 TI - Effects of charcoal kiln saunas (Jjimjilbang) on psychological states. AB - This uncontrolled intervention study explored the effects of sauna bathing utilizing residual heat from charcoal kilns (charcoal kiln saunas) on psychological states. Forty-five volunteers (24 males and 21 females; mean age 51.9 years (S.D. 15.7) visiting a bamboo charcoal kiln in Japan participated in the study. They completed a shortened version of the Profile of Mood States (POMS) and State-Trait Anxiety Inventory (STAI) before and after charcoal kiln sauna bathing in order to determine mood and anxiety states. Six factors relating to mood were measured using the POMS: Tension-Anxiety, Depression-Dejection, Anger-Hostility, Vigor, Fatigue, and Confusion. The two anxiety concepts of state anxiety and trait anxiety were also measured. Changes in psychological states before and after sauna bathing were then determined. All mood scales and both manifest anxiety measures were improved after sauna bathing. Charcoal kiln sauna bathing appears to improve mood and decrease anxiety. It is a limitation of this study that this was a descriptive prospective and an uncontrolled intervention study. Further investigation of the improvement of trait anxiety is required. PMID- 18396260 TI - Picture-plane inversion leads to qualitative changes of face perception. AB - Presenting a face stimulus upside-down generally causes a larger deficit in perceiving metric distances between facial features ("configuration") than local properties of these features. This effect supports a qualitative account of face inversion: the same transformation affects the processing of different kinds of information differently. However, this view has been recently challenged by studies reporting equal inversion costs of performance for discriminating featural and configural manipulations on faces. In this paper I argue that these studies did not replicate previous results due to methodological factors rather than largely irrelevant parameters such as having equal performance for configural and featural conditions at upright orientation, or randomizing trials across conditions. I also argue that identifying similar diagnostic features (eyes and eyebrows) for discriminating individual faces at upright and inverted orientations by means of response classification methods does not dismiss at all the qualitative view of face inversion. Considering these elements as well as both behavioral and neuropsychological evidence, I propose that the generally larger effect of inversion for processing configural than featural cues is a mere consequence of the disruption of holistic face perception. That is, configural relations necessarily involve two or more distant features on the face, such that their perception is most dependent on the ability to perceive simultaneously multiple features of a face as a whole. PMID- 18396261 TI - Contemporary research findings in shiftwork. PMID- 18396262 TI - Anxiety sensitivity, body vigilance and fear of pain. AB - The aim of the present study was to investigate the role of anxiety sensitivity (AS) as a factor relevant to pain and pain persistence. Two studies were conducted to examine the relationship between AS, body vigilance and the experience of pain in non-clinical samples. Study 1 investigated the relationship between AS and body vigilance that was operationalized by the detection latency for innocuous electrical stimuli; trait anxiety and neuroticism were also included as covariates. Results indicated that the high AS group (N=69) presented shorter detection latency than the low AS group (N=70); neuroticism and trait anxiety did not have significant effects on detection latency. Using another sample, Study 2 investigated the relationship between AS, body vigilance, pain tolerance, catastrophizing, and self-reported distress and pain during a cold pressor task. Neuroticism, trait anxiety and fear of pain were included as covariates. Results showed significant differences between high- (N=66) and low- (N=69) AS groups in body vigilance, catastrophizing and tolerance. The covariates neuroticism, trait anxiety and fear of pain did not have any significant effects. No significant differences were found in pain and distress ratings. Results from both studies support the importance of AS in body vigilance and the experience of pain. The theoretical, preventive and clinical implications of these findings are discussed. PMID- 18396263 TI - Benzo[a]pyrene inhibits osteoclastogenesis by affecting RANKL-induced activation of NF-kappaB. AB - Exposure to polycyclic aryl hydrocarbons is linked to cancer, immunosuppression and other numerous health problems. We previously demonstrated that exposure to benzo[a]pyrene (BaP), an environmental pollutant present in high concentrations in urban smog and cigarette smoke, inhibits osteoclast differentiation and bone resorption. We hypothesized that this inhibition could be due to crosstalk between the receptor activator of NF-kappaB ligand (RANKL) and AhR signaling cascades competing for NF-kappaB, a common transcription factor for both pathways. RAW264.7 cells (a mouse macrophage cell line capable of differentiating into osteoclasts in the presence of RANKL) were exposed to different concentrations of RANKL and BaP and the effect on NF-kappaB activation, nuclear translocation, as well as the effect of NF-kappaB inhibitors on BaP-mediated CYP1B1 gene expression was measured. The results demonstrated that BaP inhibited both RANKL-induced NF-kappaB activation and nuclear translocation. At the same time, BaP-induced CYP1B1 gene expression was inhibited by two NF-kappaB inhibitors in a dose-dependent manner, demonstrating that NF-kappaB is involved in a BaP-mediated signaling pathway. A reporter gene assay showed that both BaP and RANKL-induced luciferase reporter gene transcription under the control of NF kappaB response elements. Co-immunoprecipitation results demonstrated that AhR interacted with NF-kappaB p65 in RAW cells and BaP appeared to enhance this interaction. However, in the presence of RANKL, we did not observe any interaction between AhR and p65. These results support our hypothesis that BaP mediated inhibition of osteoclastogenesis is a consequence of crosstalk between AhR and RANKL signaling pathways competing for the common transcription factor NF kappaB. PMID- 18396264 TI - beta1- and beta2-adrenoceptor induced synaptic facilitation in rat basolateral amygdala. AB - The expression and characteristics of beta-adrenoceptor subtypes (beta1 and beta2) and their agonist actions on synaptic transmission in the basolateral amygdala (BLA) of the rat were examined using in situ hybridization, quantitative real-time PCR, Western blot analysis and field potential recording. In situ hybridization data revealed an intense distribution of beta1-and beta2 adrenoceptor mRNA in the BLA. Real-time PCR analysis of rat amygdala revealed significant transcriptional expression levels of both beta-adrenoceptors, with beta2-adrenoceptors outnumbering beta1-adrenoceptors in a ratio of 2.9 to 1. Bath application of the selective beta1-adrenoceptor agonist xamoterol hemifumarate (10 microM) facilitated the excitatory field synaptic potential evoked in the BLA by stimulation of the external capsule by 186.5+/-10.7% of control amplitude. In the presence of the selective beta1-adrenoceptor antagonist betaxolol hydrochloride (30 microM), the facilitating effects of field excitatory synaptic potential induced by the agonist were reduced to 126.1+/-2.3 % of control amplitude in the BLA. Bath application of the selective beta2-adrenoceptor agonist salmeterol (15 microM) facilitated the excitatory field synaptic potential evoked in the BLA by stimulation of the external capsule by 167.3+/-9.7 % of control amplitude. In the presence of the selective beta2-adrenoceptor antagonist ICI 118,551 HCl (30 microM), the facilitating effects of field excitatory synaptic potential induced by the agonist were reduced to 121.1+/-4.1 % of control amplitude in the BLA. These data suggest that beta-adrenoceptor mediated synaptic facilitation in the amygdala is mediated by both beta1 and beta2-adrenoceptor activation. PMID- 18396265 TI - Stimulation of axonal sprouting by trophic factors immobilized within the wound core. AB - Traumatic injury to the CNS results in peri-wound sprouting without significant axonal growth beyond the lesion edge. We have previously demonstrated that dopaminergic sprouting in the injured striatum follows an increasing gradient of BDNF and GDNF expression, with sprouting ceasing at the point of maximal factor expression. Progressively more complicated associations of sprouting fibers with increasingly activated microglia and macrophages suggest these factors are localized to the cell surface. To establish whether an increased concentration of immobilized BDNF and GDNF could stimulate axonal growth beyond the lesion edge, both factors were covalently attached to 10 microm polycarbonate microspheres. These spheres were implanted into the site of striatal injury 1 week after lesioning. A profusion of axons grew from the region of the lesion edge across the surface of the spheres. Some axons traversed the entire site of injury. Ultrastructural examination demonstrated the juxtaposition of regenerating axons to the surface of implanted spheres. CSPG immunostaining demonstrated that, in animals implanted with neurotrophin-microspheres, axonal growth was induced beyond the area of maximal CSPG reactivity. Surprisingly however, CSPG production at the wound edge was greater in control animals than those implanted with neurotrophin-microspheres. Overall, we show that axonal growth can be encouraged beyond the wound edge by an elevated concentration of immobilized trophic factors. This growth occurs despite the presence of inhibitory CSPGs at the lesion edge. Axonal growth appears to be stimulated mainly via the direct effects of neurotrophins. However, there also appears to be an indirect mechanism whereby neurotrophins reduce the synthesis of CSPG at the wound edge, making the peri wound environment more permissive. PMID- 18396267 TI - Laboratory diet profoundly alters gene expression and confounds genomic analysis in mouse liver and lung. AB - Nutritional studies in laboratory animals have long shown that various dietary components can contribute to altered gene expression and metabolism, but diet alone has not been considered in whole animal genomic studies. In this study, global gene expression changes in mice fed either a non-purified chow or a purified diet were investigated and background metal levels in the two diets were measured by ICP-MS. C57BL/6J mice were raised for 5 weeks on either the cereal based, non-purified LRD-5001 diet or the purified, casein-based AIN-76A diet, as part of a larger study examining the effects of low dose arsenic (As) in the diet or drinking water. Affymetrix Mouse Whole Genome 430 2.0 microarrays were used to assess gene expression changes in the liver and lung. Microarray analysis revealed that animals fed the LRD-5001 diet displayed a significantly higher hepatic expression of Phase I and II metabolism genes as well as other metabolic genes. The LRD-5001 diet masked the As-induced gene expression changes that were clearly seen in the animals fed the AIN-76A diet when each dietary group was exposed to 100 ppb As in drinking water. Trace metal analysis revealed that the LRD-5001 diet contained a mixture of inorganic and organic As at a total concentration of 390 ppb, while the AIN-76A diet contained approximately 20 ppb. These findings indicate that the use of non-purified diets may profoundly alter observable patterns of change induced by arsenic and, likely, by other experimental treatments, particularly, altering gene and protein expression. PMID- 18396266 TI - The rate of intravenous cocaine administration alters c-fos mRNA expression and the temporal dynamics of dopamine, but not glutamate, overflow in the striatum. AB - The rapid entry of drugs into the brain is thought to increase the propensity for addiction. The mechanisms that underlie this effect are not known, but variation in the rate of intravenous cocaine delivery does influence its ability to induce immediate early gene expression (IEG) in the striatum, and to produce psychomotor sensitization. Both IEG induction and psychomotor sensitization are dependent upon dopamine and glutamate neurotransmission within the striatum. We hypothesized, therefore, that varying the rate of intravenous cocaine delivery might influence dopamine and/or glutamate overflow in the striatum. To test this we used microdialysis coupled to on-line capillary electrophoresis and laser induced fluorescence, which allows for very rapid sampling, to compare the effects of a rapid (5 s) versus a slow (100 s) intravenous cocaine infusion on extracellular dopamine and glutamate levels in the striatum of freely moving rats. An acute injection of cocaine had no effect on extracellular glutamate, at either rate tested. In contrast, although peak levels of dopamine were unaffected by infusion rate, dopamine levels increased more rapidly when cocaine was administered over 5 versus 100 s. Moreover, c-fos mRNA expression in the region of the striatum sampled was greater when cocaine was administered rapidly than when given slowly. These data suggest that small differences in the temporal dynamics of dopamine neurotransmission may have a large effect on the subsequent induction of intracellular signalling cascades that lead to immediate early gene expression, and in this way influence the ability of cocaine to produce long lasting changes in brain and behavior. PMID- 18396268 TI - Salubrious effect of Kalpaamruthaa, a modified indigenous preparation in adjuvant induced arthritis in rats--a biochemical approach. AB - BACKGROUND: Interactions between the phytochemicals and drugs and their combinations are capable of providing longer remissions and perhaps a complete cure for many diseases including rheumatoid arthritis (RA). In addition to articular manifestations in RA, extra-articular signs involving reticuloendothelial and hepatic systems are an indication of more severe disease and thus, have prognostic value. OBJECTIVES: The present study was designed to illustrate the beneficial outcome of the drug Kalpaamruthaa (constituting Semecarpus anacardium nut milk extract, fresh dried powder of Emblica officinalis fruit and honey) in adjuvant-induced arthritic rat model with respect to the changes in extra-articular manifestation involving hematological and cellular constituents. MATERIAL AND METHODS: Levels of hematological parameters, cellular constituents, activities of marker enzymes and the level of DNA damage were assessed in control, arthritis-induced, SA, KA and drug control treated rats. RESULTS AND CONCLUSION: Significant decrease (p<0.005) in the levels of Hb, RBC, PCV, total protein, albumin, A/G ratio, plasma uric acid, urinary urea, uric acid, creatinine, FFA, HDL and significant increase (p<0.05) in the levels of WBC, platelet count, ESR, globulin, plasma creatinine, blood glucose, urea, AST, ALT, ALP, TC, FC, TG, PL, LDL and VLDL were observed in arthritic rats. No other significant change was observed in tissue DNA and RNA levels of control and experimental animals. On the contrary an increase in DNA damage was observed in arthritic rats when compared to control animals. The above said derangements were brought back to near normal levels upon SA and KA treatments and KA revealed a profound beneficial effect than SA. The enhanced effect of KA might be attributed to the combined effects of phytoconstituents such as flavonoids, tannins and other compounds such as vitamin C present in KA. Thus KA via this preliminary protective effect might contribute to the amelioration of the disease process. PMID- 18396269 TI - Cognitive, linguistic and affective disturbances following a right superior cerebellar artery infarction: a case study. AB - The cerebellar cognitive affective syndrome (CCAS) is a neurobehavioral syndrome that may develop after congenital and acquired cerebellar lesions. The syndrome consists of deficits in executive functioning, spatial cognition, visual-spatial memory and language and also involves personality and behavioral changes. We describe a 58-year-old right-handed man who in addition to affective disturbances presented with a unique combination of cognitive and linguistic deficits following an ischemic infarction in the vascular territory of the right superior cerebellar artery (SCA). Neurocognitive and neurolinguistic examinations were performed in the acute phase (10 days post-onset) and lesion phase (four weeks post-onset) of the stroke. A Tc-99m-ECD SPECT study was performed five weeks after the stroke. Acute phase data revealed a generalized cognitive decline and mild transcortical sensory aphasia. In the lesion phase, the neurobehavioral tableau was dominated by executive dysfunctions, disrupted divided attention, disturbed visual-spatial organization and behavioral abnormalities. Neurolinguistic investigations disclosed visual dyslexia and surface dysgraphia. Reading of words and visual lexical decision tasks of words and nonwords were severely defective and predominantly characterized by visual errors. In addition, writing irregular and ambiguous words resulted in regularization errors (phonologically plausible errors based on phoneme-grapheme correspondence rules). In the absence of any structural damage in the supratentorial brain regions, a quantified SPECT study showed a relative hypoperfusion in the right cerebellar hemisphere and the left medial frontal lobe. CCAS is for the first time reported in association with visual dyslexia and surface dysgraphia. We hypothesize that the cognitive and linguistic deficits might result from functional disruption of the cerebellar-encephalic pathways, connecting the cerebellum to the frontal supratentorial areas which subserve attentional and planning processes. This phenomenon of crossed cerebellar-cerebral diaschisis is supported by SPECT findings revealing a hypoperfusion in the anatomoclinically suspected brain regions. The constellation of cognitive, linguistic and behavioral symptoms adds new evidence to the multifaceted area of cerebellar neurocognition and demonstrates that the cerebellum might play a crucial role in cognitive, linguistic, and affective processing. PMID- 18396270 TI - Modulation of steroidogenesis and xenobiotic biotransformation responses in zebrafish (Danio rerio) exposed to water-soluble fraction of crude oil. AB - The induction of CYP enzyme activities, particularly CYP1A1, through the aryl hydrocarbon receptor (AhR) in most vertebrate species is among the most studied biochemical response to planar and aromatic organic contaminant exposure. Since P450 families play central roles in the oxidative metabolism of a wide range of exogenous and endogenous compounds, interactions between the biotransformation processes and reproductive physiological responses are inevitable. Steroidogenesis is the process by which specialized cells in specific tissues, such as the gonad, brain (neurosteroids) and kidney, synthesize steroid hormones. In the present study, we evaluated the effects of water-soluble fraction (WSF) of crude oil on the xenobiotic biotransformation and steroidogenic processes in the head (brain) and whole-body tissue of a model species by transcript analysis using quantitative (real-time) polymerase chain reaction (qPCR), enzyme activities and steroid hormone (testosterone: T and 17beta-estradiol: E2) levels using enzyme immune assay (EIA). Our data showed that exposure of fish to WSF produced an apparent concentration-specific increase of AhR1, CYP1A1 and 3beta hydroxysteroid dehydrogenase (3beta-HSD) mRNA levels, and decrease of AhR2. On the activity level, WSF produced concentration-specific increase of ethoxyresorufin O-deethylase (EROD), benzyloxyresorufin (BROD) methoxyresorufin (MROD) and pentoxyresorufin (PROD) activities in whole-body tissue. In the steroidogenic pathway, WSF exposure produced apparent concentration-specific decrease of ER* and ERbeta, steroidogenic acute regulatory (StAR) protein, cytochrome P450 side-chain cleavage (P450scc), P450aromA and P450aromB mRNA expression. For steroid hormones, while T levels decreased, E2 levels increased in an apparent WSF concentration-specific manner. In general, the xenobiotic biotransformation and estrogenic responses showed negative relationship after exposure of zebrafish to WSF, suggesting an interaction between these physiological pathways. The relationship between WSF mediated changes in brain StAR, P450scc, 3beta-HSD, ER*alpha, ERbeta, P450aromA, P450aromB and whole-body steroid hormone levels suggests that the experimental animals might be experiencing altered neurosteroidogenesis probably through increased activity level of the biotransformation system. Thus, these responses might represent sensitive diagnostic tools for short-term and acute exposure of fish or other aquatic organisms to WSF. PMID- 18396271 TI - Socioeconomic disparities in air pollution-associated mortality. AB - This study aimed to determine whether individuals with lower socioeconomic status (SES) were more susceptible to the acute effects of ambient air pollution than those with higher SES. We included 24,357 Hong Kong Chinese aged 30 or above who died of natural causes in 1998. Information on individual socioeconomic characteristics was obtained by interviewing proxy informants with a standardized questionnaire in all four death registries. Individual SES was indicated by three measures: type of housing, occupational group and education attainment. Poisson regression was performed to assess the short-term effects of ambient air pollution measured by PM(10), NO(2), SO(2) and O(3) on mortality for each SES group. The differences in the effects between SES groups were estimated by the interaction between air pollution and SES. We found that PM(10) and NO(2) were associated with greater risk of mortality on people living in public rental housing than in private housing. The effects of all four pollutants were significantly greater in blue-collar workers than the never-employed and white collar groups (p<0.05). However, we found no compelling evidence of effect modification by education attainment. Our results provide new evidence on the role of individual's SES as effect modifiers of the short-term effects of air pollution on mortality. The reduction of risks associated with air pollution for socially disadvantaged populations should be a high priority in public health and environmental policies. PMID- 18396273 TI - Hyperglycaemia as a determinant of cognitive decline in patients with type 1 diabetes. AB - Individuals with type 1 diabetes show mild performance deficits in a range of neuropsychological tests compared to healthy controls, but the mechanisms underlying this cognitive deterioration are still poorly understood. Basically, two diabetes-related mechanisms can be postulated: recurrent severe hypoglycaemia and/or chronic hyperglycaemia. Intensive insulin therapy in type 1 diabetes, resulting in a durable improvement of glycaemic control, has been shown to lower the risk of long-term microvascular and macrovascular complications. The down side of striving for strict glycaemic control is the considerably elevated risk of severe hypoglycaemia, sometimes leading to seizure or coma. While retrospective studies in adult patients with type 1 diabetes have suggested an association between a history of recurrent severe hypoglycaemia and a modest or even severe degree of cognitive impairment, large prospective studies have failed to confirm this association. Only fairly recently, better appreciation of the possible deleterious effects of chronic hyperglycaemia on brain function and structure is emerging. In addition, it can be hypothesized that hyperglycaemia associated microvascular changes in the brain are responsible for the cognitive decline in patients with type 1 diabetes. This review presents various pathophysiological considerations concerning the cognitive decline in patients with type 1 diabetes. PMID- 18396272 TI - Increased adiposity on normal diet, but decreased susceptibility to diet-induced obesity in mu-opioid receptor-deficient mice. AB - The mu-opioid receptor encoded by the Oprm1 gene plays a crucial role in the mediation of food reward and drug-induced positive reinforcement, but its genetic deletion has been shown to provide food intake-independent, partial protection from diet-induced obesity. We hypothesized that mu-opioid receptor-deficient mice would show an even greater, intake-dependent, resistance to high-fat diet-induced obesity if the diet comprises a sweet component. We generated an F2 population by crossing the heterozygous offspring of homozygous female Oprm1(-/-) mice (on a mixed C57BL/6 and BALB/c genetic background) with male inbred C57BL/6 mice. Groups of genotyped wild-type (WT) and homozygous mutant (KO) males and females were fed either control chow or a high caloric palatable diet consisting of sweet, liquid chocolate-flavored Ensure together with a solid high-fat diet. Food intake, body weight, and body composition was measured over a period of 16 weeks. Unexpectedly, male, and to a lesser extent female, KO mice fed chow for the entire period showed progressively increased body weight and adiposity while eating significantly more chow. In contrast, when exposed to the sweet plus high fat diet, male, and to a lesser extent female, KO mice gained significantly less body weight and fat mass compared to WT mice when using chow fed counterparts for reference values. Male KO mice consumed 33% less of the sweet liquid diet but increased intake of high-fat pellets, so that total calorie intake was not different from WT animals. These results demonstrate a dissociation of the role of mu-opioid receptors in the control of adiposity for different diets and sex. On a bland diet, normal receptor function appears to confer a slightly catabolic predisposition, but on a highly palatable diet, it confers an anabolic metabolic profile, favoring fat accretion. Because of the complexity of mu-opioid gene regulation and tissue distribution, more selective and targeted approaches will be necessary to fully understand the underlying mechanisms. PMID- 18396274 TI - Expression of the myodystrophic R453W mutation of lamin A in C2C12 myoblasts causes promoter-specific and global epigenetic defects. AB - Autosomal dominant Emery-Dreifuss muscular dystrophy (EDMD) is characterized by muscle wasting and is caused by mutations in the LMNA gene encoding A-type lamins. Overexpression of the EDMD lamin A R453W mutation in C2C12 myoblasts impairs myogenic differentiation. We show here the influence of stable expression of the R453W and of the Dunnigan-type partial lipodystrophy R482W mutation of lamin A in C2C12 cells on transcription and epigenetic regulation of the myogenin (Myog) gene and on global chromatin organization. Expression of R453W-, but not R482W-lamin A, impairs activation of Myog and maintains a repressive chromatin state on the Myog promoter upon induction of differentiation, marked by H3 lysine (K) 9 dimethylation and failure to hypertrimethylate H3K4. Cells expressing WT LaA also fail to hypertrimethylate H3K4. No defect occurs at the level of Myog promoter DNA methylation in any of the clones. Expression of R453W-lamin A and to a lesser extent R482W-lamin A in undifferentiated C2C12 cells redistributes H3K9me3 from pericentric heterochromatin. R453W-lamin A also elicits a redistribution of H3K27me3 from inactive X (Xi) and partial decondensation of Xi, but maintains Xist expression and coating of Xi, indicating that Xi remains inactivated. Our results argue that gene-specific and genome-wide chromatin rearrangements may constitute a molecular basis for laminopathies. PMID- 18396276 TI - Biomechanical properties of human and porcine corneas. AB - The suitability of porcine corneas as approximate models for human corneas in mechanical property characterisation studies is experimentally assessed. Thirty seven human donor corneas and thirty four ex-vivo porcine corneas were tested under inflation conditions to determine their short-term stress-strain behaviour and long-term creep behaviour up to 2.8 h (10,000 s). Vertical strips extracted from further 12 human corneas and 10 porcine corneas were subjected to stress relaxation tests for up to 20 min at different stress levels. Human and porcine corneas were observed to have almost the same form of behaviour under short and long-term loading. They both exhibited non-linear stress-strain behaviour and reacted to sustained loading in a similar fashion. However, human corneas were significantly stiffer than porcine corneas. They also crept less under long-term loading and could sustain their stress state for longer compared to porcine corneas. These differences, in addition to others identified earlier in relation to corneal mechanical anisotropy, cast doubt on the suitability of porcine corneas as models for human corneas in mechanical studies. PMID- 18396275 TI - Structural requirements for the assembly of LINC complexes and their function in cellular mechanical stiffness. AB - The evolutionary-conserved interactions between KASH and SUN domain-containing proteins within the perinuclear space establish physical connections, called LINC complexes, between the nucleus and the cytoskeleton. Here, we show that the KASH domains of Nesprins 1, 2 and 3 interact promiscuously with luminal domains of Sun1 and Sun2. These constructs disrupt endogenous LINC complexes as indicated by the displacement of endogenous Nesprins from the nuclear envelope. We also provide evidence that KASH domains most probably fit a pocket provided by SUN domains and that post-translational modifications are dispensable for that interaction. We demonstrate that the disruption of endogenous LINC complexes affect cellular mechanical stiffness to an extent that compares to the loss of mechanical stiffness previously reported in embryonic fibroblasts derived from mouse lacking A-type lamins, a mouse model of muscular dystrophies and cardiomyopathies. These findings support a model whereby physical connections between the nucleus and the cytoskeleton are mediated by interactions between diverse combinations of Sun proteins and Nesprins through their respective evolutionary-conserved domains. Furthermore, they emphasize, for the first time, the relevance of LINC complexes in cellular mechanical stiffness suggesting a possible involvement of their disruption in various laminopathies, a group of human diseases linked to mutations of A-type lamins. PMID- 18396277 TI - Mutation of Dock5, a member of the guanine exchange factor Dock180 superfamily, in the rupture of lens cataract mouse. AB - Rupture of lens cataract (RLC) in the mouse is a spontaneous mutation inherited by a single autosomal recessive gene mapped on chromosome 14. Fine mapping of the mutant locus revealed a nucleotide deletion of 27-bp at the end of 15th exon of Dock5 (Dedicator of cytokinesis-5), a member of the Dock gene superfamily. Since the deletion occurred in-frame, the RLC-DOCK5 protein had a deletion of 9 amino acids (a.a. 506-514) in the DHR1 (DOCK homology region-1) domain that is essential for DOCK5, a GTP-exchanger for Rac1. Although Dock5 mRNA was intensely expressed equally in mutant and wild-type lenses, DOCK5 protein was hardly detectable in the mutant lens. In contrast, expression of Dock180, another member of Dock subfamily A, was not affected in RLC. Immunohistochemically, DOCK5 was stained intensely in the cytoplasm of the anterior epithelial cells and weakly in lens fiber of the wild type lenses, but little in RLC lens. These observations suggest that the mutation may somehow destabilize DOCK5 protein. We propose to designate the mutant allele of rlc as Dock5rlc. Relevance of the signaling pathway involving DOCK5-RAC1 in maintenance of lens integrity of growing lens is discussed. PMID- 18396278 TI - Chick embryonic Schwann cells migrate anodally in small electrical fields. AB - Little is known about the cues that guide migrating neural crest derivatives to their targets. This lack of understanding is especially significant in the case of Schwann cells, which have been transplanted into the central nervous system in an effort to promote axonal myelination after injury or disease. We have investigated the response of Schwann cells, cultured from the peripheral nerves of E7/8 chick embryos, to applied electrical fields. We find that they respond by migrating to the anode, and show a significant anodal bias in directionality at 3 mV mm(-1). This is the smallest electrical field that has been shown to affect cellular movement or growth in culture, and the anodal direction is surprising given the known cathodal responses of neural crest cells. The effective fields are considerably smaller than endogenous electrical fields that have been measured in embryonic tissues. PMID- 18396279 TI - Neonatal tryptophan dietary restriction alters development of retinotectal projections in rats. AB - The specification of sensory neural circuits includes the elimination of transitory axon collaterals/synapses that takes place during early post natal life, an important step for the acquisition of topographical order of sensory systems. Serotonin has been implicated in the patterning of connections in subcortical and cortical circuits. We investigated the effects of the dietary restriction of the only serotonin precursor, tryptophan, on the development of the uncrossed retinotectal pathway in pigmented rats. Litters were fed through their mothers with either a tryptophan restricted, corn and gelatin based diet or a similar control diet complemented with tryptophan during the lactation period. The developmental status of the uncrossed retinotectal terminal fields was studied after the anterograde transport of horseradish peroxidase injected into one eye. We also studied the effects of tryptophan restriction on 5-HT immunoreactivity of raphe neurons, on cAMP levels in the visual layers of the superior colliculus and on protein synthesis among retinal neurons. We found that tryptophan restriction resulted in reduced weight gain among tryptophan restricted rats, without differences in protein synthesis between tryptophan complemented and restricted groups. Tryptophan restriction was also associated with a reduction of serotonin immunoreactive cells in the raphe nuclei and increased cAMP levels in the superior colliculus. Finally we found that neonatal tryptophan restriction resulted in an abnormal patterning of retinotectal topography, which was consistent with a developmental delay in axonal elimination and fine tuning of central connections. These results suggest, therefore, that dietary tryptophan is crucial for the influence of serotonin in the maturation of central visual connections. PMID- 18396281 TI - Entamoeba invadens: influence of 60 Hz magnetic fields on growth and differentiation. AB - Exposure to extremely low-frequency (ELF) electromagnetic fields appears to result in a number of important biological changes. In the present study, we evaluated the effects of 60 Hz sinusoidal magnetic fields (MF) at magnetic flux densities of 1.0, 1.5 and 2.0 mT on growth and differentiation of the protozoan Entamoeba invadens. We demonstrated an inhibitory growth effect when trophozoite cultures were exposed to 1.5 and 2.0 mT. Furthermore, we found that there was not a synergistic effect in cultures co-exposed to MF and Metronidazole, a cytotoxic drug against amoebic cells. In addition, MF exposure inhibited the encystation process of E. invadens. PMID- 18396280 TI - Functional MRI and other non-invasive imaging technologies: providing visual biomarkers for spinal cord structure and function after injury. AB - Substantial progress has been made towards understanding the molecular basis for limited endogenous central nervous system (CNS) axonal growth after injuries such as spinal cord trauma. Realization of the potential benefit of therapeutic interventions requires methods to assess axonal growth and functional reorganization over time after neurological damage. Here, we discuss the technical challenges of analyzing and interpreting the effects of various interventions on CNS repair, specifically in the context of spinal cord injury. Evolving technologies such as functional magnetic resonance imaging and other non invasive imaging techniques will be reviewed. These technologies should revolutionize our ability to track changes in both CNS structure and function. PMID- 18396283 TI - Experimental studies on Echinococcus multilocularis in Japan, focusing on biohazardous stages of the parasite. AB - Alveolar echinococcosis (AE), caused by the active growth of larval Echinococcus multilocularis mostly in the liver, is usually fatal zoonotic disease if not adequately treated. Humans become infected via oral ingestion of the parasite eggs, which are thus biohazardous to humans and should be handled under restricted conditions. In this review, we present findings in experimental studies mainly performed at a safety facility in Japan, examining the biohazadous stages of the parasite (Hokkaido isolate) including its egg and adult worm stages. This article deals mainly with the parasite development in various experimental and wild animals, environmental factors affecting viability of the parasite eggs, and molecular biological studies on adult worms. The findings shown herein have provided a basis to better understand basic biology and natural transmission of E. multilocularis in Hokkaido, a highly endemic area of AE in northern Japan, and also to establish effective preventive measures against the disease. PMID- 18396282 TI - Counter-regulatory anti-parasite cytokine responses during concurrent Plasmodium yoelii and intestinal helminth infections in mice. AB - Malaria and helminth infections are two of the most prevalent parasitic diseases globally. While concomitant infection is common, mechanisms contributing to altered disease outcomes during co-infection remain poorly defined. We have previously reported exacerbation of normally non-lethal Plasmodium yoelii malaria in BALB/c mice chronically infected with the intestinal trematode Echinostoma caproni. The goal of the present studies was to determine the effect of helminth infection on IFN-gamma and other key cytokines during malaria co-infection in the P. yoelii-E. caproni and P. yoelii-Heligmosomoides polygyrus model systems. Polyclonally stimulated spleen cells from both E. caproni- and H. polygyrus infected mice produced significantly lower amounts of IFN-gamma during P. yoelii co-infection than malaria-only infected mice. Furthermore, the magnitude of IFN gamma suppression was correlated with the relative amounts of IL-4 induced by these helminths (E. caproni=low; H. polygyrus=high), but not IL-10. Concurrent malaria infection also suppressed helminth-associated IL-4 responses, indicating that immunologic counter-regulation occurs during co-infection with malaria and intestinal helminths. PMID- 18396284 TI - Iron storage is significantly increased in peritoneal macrophages of endometriosis patients and correlates with iron overload in peritoneal fluid. AB - OBJECTIVE: To further investigate peritoneal iron disruption in endometriosis by studying iron storage in peritoneal macrophages of patients with endometriosis compared with controls. DESIGN: Cross-sectional study. SETTING: Academic gynecology research unit in a university hospital. PATIENT(S): Fifty patients undergoing laparoscopy. INTERVENTION(S): Collection of peritoneal fluid samples (N = 50) from patients with (n = 27) and without (n = 23) endometriosis undergoing laparoscopy. MAIN OUTCOME MEASURE(S): Quantification of peritoneal macrophage ferritin by immunocytochemical staining and immunodensitometry and measurement of peritoneal iron, transferrin, ferritin, and prohepcidin concentrations. RESULT(S): The optical density of peritoneal macrophage ferritin staining was statistically significantly higher in endometriosis patients than in controls. Higher iron concentrations, transferrin saturations, and ferritin concentrations were also detected in case of endometriosis. A statistically significant positive correlation was found between the optical density of macrophage ferritin staining and peritoneal iron concentrations in endometriosis and control patients. CONCLUSION(S): Iron storage is statistically significantly increased in peritoneal macrophages of patients with endometriosis and correlates with iron overload in peritoneal fluid. The potential implications of iron accumulation in peritoneal macrophages in case of endometriosis are discussed. PMID- 18396285 TI - Evaluation of posttranslational processing of proopiomelanocortin in the banded houndshark pituitary by combined cDNA cloning and mass spectrometry. AB - Proopiomelanocortin (POMC) is cleaved into small peptides, such as adrenocorticotropic hormone (ACTH), melanocyte-stimulating hormones (MSHs), and beta-endorphin (beta-END), by tissue-specific posttranslational processing in the corticotrophs of the pars distalis (PD) and melanotrophs of the neurointermediate lobe (NIL) of the pituitary. We examined the posttranslational processing of POMC in the pituitary of the banded houndshark Triakis scyllium by molecular cloning and subsequent mass spectrometric identification of the POMC-derived peptides in the pituitary extracts. One-fifth of the randomly selected clones from a Triakis pituitary cDNA library contained a cDNA encoding for POMC. Triakis prePOMC contained 4 MSHs and a single beta-END, as has been observed in case of other cartilaginous fish POMCs. These predicted hormonal segments were flanked by basic amino acid residues, which are the cleavage sites for the processing enzymes, i.e., protein convertases. Mass spectrometry was performed using PD (including most parts of the rostral and proximal PD) and NIL extracts to detect mass values corresponding to the POMC-derived peptides. Consequently, ACTH, beta-END, and the joining peptide (JP) were detected in the PD extract, while MSHs, processed beta END, and some other POMC-derived peptides were identified in the NIL extract; however, neither acetylated alpha-MSH nor acetylated beta-END was detected in the latter. These tissue-specific POMC processing patterns are similar to those of the other vertebrate pituitaries; however, the absence of acetylated peptides suggests the lack of an acetylation system in the melanotrophs in the NIL of the Triakis pituitary. PMID- 18396286 TI - The enigma of the Sangiran 4 palate revisited. AB - The Sangiran 4 palate has been controversial since its discovery in the 1930s because it retains a number of more primitive morphologies such as projecting canines and precanine diastemata. These characters have led some workers to question the hominid status of the palate, suggesting that it is both too large and too primitive to belong to the same individual as the Sangiran 4 cranial fragments. The palate has instead been diagnosed as a new species of Pongo. The present study re-evaluates this controversy through the analysis of new metric data and comparisons with more recently published fossil discoveries. This analysis shows that the Sangiran 4 palate is not unique, and shares several of these putative pongid traits with other Javan hominid fossils as well as recently described hominid specimens from Dmanisi, Georgia. These results suggest that the evolution of the earliest Asians was more complex than has previously been appreciated. PMID- 18396287 TI - Enthesis bilateral asymmetry in humans and African apes. AB - Entheses (skeletal muscle and tendon attachment sites) have often been used to infer handedness and activity variability among human populations. However, the specific roles that intensity vs. frequency of muscle contractions play in modifying entheses are not well understood and the assumption that entheses reflect muscle activity levels has been challenged. This study explores the effect of habitual muscular activity on enthesis morphology in humans and African apes by investigating bilateral asymmetry in the forelimbs and hindlimbs of these taxa. Humans have generally more developed entheses in the lower limb while African apes have generally more developed entheses in the forelimbs. All species studied have more asymmetric forelimbs than hindlimbs except humans that show more asymmetrical expression of bony spurs in the lower limbs than in the upper limbs. When comparing species, humans are always more asymmetric in ethesis development than apes for both the forelimbs and hindlimbs, which reflects the relatively greater asymmetry in limb use in humans and the more symmetric use in apes. Enthesis development may reflect cross-symmetry patterns in humans and, more subtly, a moderate handedness in apes during manipulative activities. This study suggests that enthesis morphology provides information on muscle activity levels, with greater development of entheses associated with more habitual or powerful muscle use. The general similarity of ape and human responses to muscle activity suggests that muscle activity influenced enthesis development in Plio Pleistocene hominins and that interpretation of muscle markings in these fossils can provide data for functional inferences in these extinct species. PMID- 18396288 TI - Focused grooming networks and stress alleviation in wild female baboons. AB - We examine the relationship between glucocorticoid (GC) levels and grooming behavior in wild female baboons during a period of instability in the alpha male rank position. All females' GC levels rose significantly at the onset of the unstable period, though levels in females who were at lower risk of infanticide began to decrease sooner in the following weeks. Three factors suggest that females relied on a focused grooming network as a coping mechanism to alleviate stress. First, all females' grooming networks became less diverse in the weeks following the initial upheaval. Second, females whose grooming had already focused on a few predictable partners showed a less dramatic rise in GC levels than females whose grooming network had been more diverse. Third, females who contracted their grooming network the most experienced a greater decrease in GC levels in the following week. We conclude that close bonds with a few preferred partners allow female baboons to alleviate the stress associated with social instability. PMID- 18396289 TI - A simple and rapid method for electromagnetic field distortion correction when using two Fastrak sensors for biomechanical studies. AB - The article presents a simple and rapid method for the correction of electromagnetic distortions when using electromagnetic Fastrak (Polhemus, USA) sensors. It is based on the minimization of objective functions composed of derivative polynomial functions, hence estimating the distortion of the electromagnetic field. The polynomial functions composing the objective function each contain 35 deformation coefficients. These coefficients are then used to correct the electromagnetic measures in position and orientation. Preliminary results on the efficacy of the method are presented for two subjects who walked on a treadmill, and for whom relative movement of the lower leg with respect to the thigh was recorded using two Fastrak sensors. The corrected Fastrak measurements were compared with optoelectronic measurements (Vicon, USA), which are not affected by distortions as electromagnetic sensors are. Results showed that after 3 min of calibrating a volume of approximately 1m(3), the method proved to be efficient in correcting errors in orientation (56% (2.72-1.12 degrees ), 78% (4.4-0.89 degrees ), and 56% (2.25-0.90 degrees ) of error reduction in the respective flexion/extension, ab/adduction and tibial internal/external rotation) and position (53% (18.9-8.9 mm), 21% (6.6-4.6mm), and 48% (15.9-8.1mm) of error reduction in the respective medial/lateral, anterior/posterior and proximal/distal translations) (values are overall means for two subjects and four calibration procedures). That amount of correction compared favorably with values presented in the literature. PMID- 18396290 TI - Viscoelastic properties of passive skeletal muscle in compression: stress relaxation behaviour and constitutive modelling. AB - The compressive properties of skeletal muscle are important in impact biomechanics, rehabilitation engineering and surgical simulation. However, the mechanical behaviour of muscle tissue in compression remains poorly characterised. In this paper, the time-dependent properties of passive skeletal muscle were investigated using a combined experimental and theoretical approach. Uniaxial ramp and hold compression tests were performed in vitro on fresh porcine skeletal muscle at various rates and orientations of the tissue fibres. Results show that above a very small compression rate, the viscoelastic component plays a significant role in muscle mechanical properties; it represents approximately 50% of the total stress reached at a compression rate of 0.5% s(-1). A stiffening effect with compression rate is observed especially in directions closer to the muscle fibres. Skeletal muscle viscoelastic behaviour is thus dependent on compression rate and fibre orientation. A model is proposed to represent the observed experimental behaviour, which is based on the quasi-linear viscoelasticity framework. A previously developed strain-dependent Young's Moduli formulation was extended with Prony series to account for the tissue viscoelastic properties. Parameters of the model were obtained by fitting to stress-relaxation data obtained in the muscle fibre, cross-fibre and 45 degrees directions. The model then successfully predicted stress-relaxation behaviour at 60 degrees from the fibre direction (errors <25%). Simultaneous fitting to data obtained at compression rates of 0.5% s(-1), 1%s(-1) and 10% s(-1) was performed and the model provided a good fit to the data as well as good predictions of muscle behaviour at rates of 0.05% s(-1) and 5% s(-1) (errors <25%). PMID- 18396291 TI - How do low horizontal forces produce disproportionately high torques in human locomotion? AB - Although horizontal ground forces are only approximately 15% of vertical forces, they account for 47% and 33% of the metabolic cost in walking and running. To explain these disproportionately high metabolic costs, we hypothesized that low horizontal ground forces generate relatively high torques on body segments during locomotion and this is mediated by long moment arms. We compared external force moment arms and discreet torques applied to the body segments by horizontal and vertical forces during walking and running. Sixteen subjects (21.9+/-1.9 years) walked at 1.5m/s and ten subjects (23.2+/-2.0 years) ran at 3.83 m/s. Segmental torques in the sagittal plane were partitioned into components due to horizontal and vertical forces and quantified by their angular impulses. The mean (+/-S.E.) ratios of horizontal to vertical ground forces (GF ratio) and angular impulses (AI ratio) in walking were 0.131 (+/-0.003, 95% confidence interval (CI) 0.124 0.137) and 0.530 (+/-0.018, CI 0.497-0.569). Results were similar in running. In both gaits the AI ratios were significantly greater than the GF ratios because the respective CI's did not overlap. The horizontal forces produced 53% and 41% as much angular impulse on the body segments, as did the vertical forces in walking and running despite being only 13% as large. In the two movements the moment arms for the horizontal forces averaged across foot, leg, thigh, and trunk body segments were 3.8 fold larger than those for the vertical forces. The data supported the hypothesis and suggest that the relatively low horizontal vs. vertical forces accounted for a disproportionately higher percentage of the angular impulses placed on the body segments and this effect was due to relatively long moment arms for horizontal forces. These results partially explain the relatively large metabolic cost of generating relatively low horizontal forces. PMID- 18396292 TI - Improving joint torque calculations: optimization-based inverse dynamics to reduce the effect of motion errors. AB - The accuracy of joint torques calculated from inverse dynamics methods is strongly dependent upon errors in body segment motion profiles, which arise from two sources of noise: the motion capture system and movement artifacts of skin mounted markers. The current study presents a method to increase the accuracy of estimated joint torques through the optimization of the angular position data used to describe these segment motions. To compute these angular data, we formulated a constrained nonlinear optimization problem with a cost function that minimizes the difference between the known ground reaction forces (GRFs) and the GRF calculated via a top-down inverse dynamics solution. To evaluate this approach, we constructed idealized error-free reference movements (of squatting and lifting) that produced a set of known "true" motions and associated true joint torques and GRF. To simulate real-world inaccuracies in motion data, these true motions were perturbed by artificial noise. We then applied our approach to these noise-induced data to determine optimized motions and related joint torques. To evaluate the efficacy of the optimization approach compared to traditional (bottom-up or top-down) inverse dynamics approaches, we computed the root mean square error (RMSE) values of joint torques derived from each approach relative to the expected true joint torques. Compared to traditional approaches, the optimization approach reduced the RMSE by 54% to 79%. Average reduction due to our method was 65%; previous methods only achieved an overall reduction of 30%. These results suggest that significant improvement in the accuracy of joint torque calculations can be achieved using this approach. PMID- 18396293 TI - Stem cells for cardiovascular repair - the challenges of the aging heart. AB - The discovery of extracardiac progenitor cells and resident cardiac stem cells in recent years has led to a great deal of interest in the development of therapeutic strategies that target these endogenous cell sources for promotion of cardiovascular repair mechanisms in the diseased heart. Cardiovascular risk increases with age and among many factors, the age-associated decline in cardiac and vascular regenerative capacity may contribute to the progressive deterioration of cardiovascular health. Thus, understanding the mechanisms which underlie the dysregulation of cardiac stem and progenitor cells may lead to the identification of novel targets and approaches to reverse this decline. In this review, we outline the current knowledge about cardiac stem and progenitor cells, their contribution to cardiovascular regenerative processes and factors that may affect their decreased function in aging individuals. Moreover, we describe the therapeutic strategies that are currently being tested in clinical trials as well as potential new avenues of investigation for the future. PMID- 18396294 TI - Genetic analysis of tumor necrosis factor-alpha (TNF-alpha) G-308A and Saitohin Q7R polymorphisms with Alzheimer's disease. AB - Neuroinflammation and abnormal phosphorylation of TAU proteins have been implicated in the etiology of Alzheimer's disease (AD). Several studies have suggested the G-308A promoter polymorphism in one of the proinflammatory cytokine genes tumor necrosis factor-alpha (TNF-alpha) encoding TNF-alpha may be associated with AD pathogenesis. Association between the Q7R polymorphism in saitohin (STH), a gene nested within the intron of the Tau gene, has also been reported. To determine whether these two polymorphisms contribute to the risk for late-onset AD (LOAD) in Chinese, we have investigated 207 sporadic LOAD patients and 222 healthy controls. The associations of the AA genotype and A-allele with LOAD (chi(2) = 8.74, df = 1, P = 0.0031, and chi(2) = 4.47, df = 1, P = 0.035) were found. After stratifying by apolipoprotein E allele 4 (APOE epsilon4) status, increased LOAD risks associated with the AA genotype and A-allele only in the APOE epsilon4 non-carriers (chi(2) = 9.21, df = 1, P = 0.002; chi(2) = 10.02, df = 1, P = 0.0015) were seen. These results suggested that the TNF-alpha gene G 308A polymorphism might be a risk factor for LOAD and dependent on APOE epsilon4 status in Chinese. Homozygous Q/Q of STH Q7R polymorphism was the only one genotype found in either LOAD group or controls. No R allele was detected in LOAD and control groups. The extremely rare frequency of the ancestral R allele differs sharply from that observed in studies in the Caucasian population, suggesting obvious ethnic differences. PMID- 18396295 TI - Aging, evolvability, and the individual benefit requirement; medical implications of aging theory controversies. AB - There is a class of theories of aging (variously termed adaptive aging, aging by design, aging selected for its own sake, or programmed death theories) that hold that an organism design that limits life span conveys benefits and was selected specifically because it limits life span. These theories have enjoyed a resurgence of popularity because of the discovery of genes that promote aging in various organisms. However, traditional evolution theory has a core tenet that excludes the possibility of evolving and retaining an individually adverse organism design, i.e. a design characteristic that reduces the ability of individual organisms to survive or reproduce without any compensating individual benefit. Various theories of aging dating from the 1950s and based on traditional evolution theory enjoy substantial popularity. Therefore, any theorist proposing an adaptive theory of aging must necessarily also propose some adjustment to traditional evolution theory that specifically addresses the individual benefit issue. This paper describes an adaptive theory of aging and describes how one of the proposed adjustments (evolvability theory) supports adaptive aging. This issue is important because adaptive theories are generally more optimistic regarding prospects for medical intervention in the aging process and also suggest different approaches in achieving such intervention. PMID- 18396296 TI - Height, weight, body mass index and offspring sex at birth in contemporary Finnish women. PMID- 18396297 TI - DNA A-tracts bending: polarization effects on electrostatic interactions across their minor groove. AB - Bending by the DNA A-tracts constitutes a contentious issue, suggesting deficiencies in the physics employed so far. Here, we inquire as to the importance in this bending of many-body polarization effects on the electrostatic interactions across their narrow minor groove. We have done this on the basis of the findings of Jarque and Buckingham who developed a procedure based on a Monte Carlo simulation for two charges of the same sign embedded in a polarizable medium. Remarkably, the present analysis reveals that for compact DNA conformations, which result from dynamic effects, an overall attractive interaction operates between the phosphate charges; this interaction is especially strong for the narrow minor groove of the A-tracts, suggesting a tendency for DNA to bend toward this groove. This tendency is in agreement with the conclusions of electrophoretic and NMR solution studies. The present analysis is also consistent with the experimental observations that the minor groove is much more easily compressible than the major groove and the bending propensity of the A-tracts is greatly reduced at "premelting" temperatures. By contrast, the dielectric screening model predicts a repulsion between the phosphate charges and is not consistent with the aforementioned bending tendency or experimental observations. PMID- 18396298 TI - Measurement of environmental trace-metal levels in Mediterranean coastal areas with transplanted mussels and DGT techniques. AB - Mussels (Mytilus galloprovincialis) and diffusive gradients in thin films (DGT) devices were deployed together for three one-month periods in coastal waters of Sardinia (Italy), in order to assess the levels of Cd, Cu, Ni, and Pb in areas differently affected by anthropogenic activities. DGT devices were more sensitive than mussels in detecting differences in metal concentrations between sites, while interpretation of the biomonitoring data revealed difficulties related to the biotic and abiotic factors that can influence the measurements. Regression analysis showed a significant correlation between Cd and Pb concentrations measured in the mussel tissues and bioavailable metal levels in seawater. Moreover transplanted mussels did not accumulate Cu and Ni following pre exposure, although DGT showed significant concentrations of bioavailable forms of these metals in seawater. The results provided an estimate of the water quality with respect to dissolved metals and pointed out the usefulness of a combination of biomonitoring and DGT techniques for a better understanding of trace metal availability in coastal waters. PMID- 18396299 TI - Effects of 4-nonylphenol exposure in mussels (Mytilus galloprovincialis) and crabs (Carcinus aestuarii) with particular emphasis on vitellogenin induction. AB - Since it is often difficult to estimate possible adverse effects due to contamination in selected ecosystems, multi-species biomonitoring may provide more information, taking into account different routes of exposure, ecological roles and metabolic capabilities of animals. In this context, we exposed for 7 days the mussel Mytilus galloprovincialis and the crab Carcinus aestuarii to 4 nonylphenol (NP), a well-known xenoestrogen. In mussels (0-0.2 mg NP l(-1)), we measured NP bioaccumulation in soft tissues and vitellogenin (Vg)-like protein levels in digestive glands from both males and females by the alkali-labile phosphate assay (ALP). As no reference data were available for crab exposure, the NP 96-h LC(50) value was previously determined. Then, in sublethally exposed (0 1.0 mg NP l(-1)) male crabs, NP bioaccumulation and Vg levels were measured in hemolymph, gonads and digestive gland. Bioaccumulation of NP increased from 43 to 371 microg g(-1) d.w. in mussels, and from 3.6 to 37 microg g(-1) d.w. in crabs, depending on the NP concentration in water. Dose-dependent Vg-like protein induction was observed in both species, appearing to be related to NP bioaccumulation, although a partial decrease was recorded at the highest concentration tested. A similar trend was observed in both digestive gland and gonad of exposed crabs; Vg increased to a lesser extent, although significantly, in hemolymph. Results demonstrated that NP induces Vg synthesis both in male and female mussels, as well as in male crabs. On the basis of the responsiveness of both species investigated, a multi-species approach is indicated in biomonitoring programmes. PMID- 18396300 TI - 5HT2 receptor activation facilitates P2X receptor mediated excitatory neurotransmission to cardiac vagal neurons in the nucleus ambiguus. AB - Parasympathetic preganglionic cardiac vagal neurons (CVNs) which dominate the control of heart rate are located within the nucleus ambiguus (NA). Serotonin (5HT), and in particular 5HT2 receptors, play an important role in cardiovascular function in the brainstem. However, there is a lack of information on the mechanisms of action of 5HT2 receptors in modulating parasympathetic cardiac activity. This study tests whether activation of 5HT2 receptors alters excitatory glutamatergic and purinergic neurotransmission to CVNs. Application of alpha methyl-5-hydroxytryptamine (alpha-Me-5HT), a 5HT2 agonist, reversibly increased both the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) in CVNs. Similar responses were obtained with alpha-methyl-5-(2 thienylmethoxy)-1H-indole-3-ethanamine hydrochloride (BW723C86), and m chlorophenylpiperazine (m-CPP), 5HT2B and 5HT2B/C receptor agonists, respectively. The facilitation evoked by alpha-Me-5HT was prevented by the 5HT2B/C receptor antagonist SB206553 hydrochloride (SB206553). Interestingly, the blockage of both NMDA and non-NMDA glutamatergic receptors did not prevent alpha Me-5HT-evoked facilitation of mEPSCs, however, the responses were blocked by the P2 receptor antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS). The responses evoked by alpha-Me-5HT were mimicked by application of alpha,beta-methylene ATP (alpha,beta-Me-ATP), a P2X receptor agonist, which were also blocked by PPADS. In summary, these results indicate activation of 5HT2 receptors facilitates excitatory purinergic, but not glutamatergic, neurotransmission to CVNs. PMID- 18396301 TI - Distinct and independent correlates of attention and awareness in a hemianopic patient. AB - Attention and awareness are intimately related concepts. Nevertheless, the two phenomena are empirically dissociable: visuo-spatial attention can act in the absence of visual awareness. We used magnetoencephalography (MEG) to record cortical neural activity from hemianopic patient GY while he performed a peripheral orientation-discrimination task in the context of an attention-cueing paradigm. The luminance contrast of target stimuli was set at GY's threshold for reports of awareness (a feeling "that something happened" in his blind visual field). GY's accuracy was significantly greater than chance and comparable, with or without awareness. GY was significantly faster to respond correctly on valid cue versus invalid-cue trials, even in the absence of awareness, confirming the action of visuo-spatial attention in the absence of awareness. Time-frequency analysis of spectral power in the gamma frequency range (30-90Hz), averaged over left parieto-occipital sensors, revealed effects of cue-validity independent of reported awareness, and effects of awareness independent of cue-validity. GY's reports of awareness were not, however, independent of his pre-target attentional state (gamma-band response to the central cue), consistent with a one-way, but not a two-way, dissociation between attention and awareness. Our evidence suggests that the observed cue-validity effect is an awareness-independent involuntary re-orienting response, and that the neurodynamics underlying the exogenous capture of attention are similar with or without awareness. The finding of a significant awareness-independent effect in the area of 40Hz implies that a stimulus-induced modulation of power in the canonical gamma band is not a sufficient condition for sensory awareness. PMID- 18396302 TI - Class targeted metabolomics: ESI ion trap screening methods for glucosinolates based on MSn fragmentation. AB - Glucosinolates are naturally occurring anionic secondary plant metabolites incorporating a thioglucosidic link to the carbon of a sulphonated oxime. There are a large number of naturally occurring glucosinolates and they are found in relatively large quantities in many plant species within the family Crucifereae. These metabolites are of interest for both their anticancer and flavour properties and in the study of nitrogen and sulphur metabolism in model plants such as Arabidopsis. Parent ion mapping is an analytical mass spectrometry approach that allows rapid assessment of glucosinolate content. Ion mapping proved to be highly sensitive and the glucosinolate sinigrin could be detected at three parts per trillion. This method takes advantage of the glucosinolate anion fragmentation which consistently produces a sulphonate ring-opened glucose moiety in the ion trap mass spectrometer, m/z 259. An intramolecular transfer mechanism for this fragmentation is presented here for the first time. This fragmentation can be exploited as a general identifier of the glucosinolate class of metabolites in plant extracts and in LCMSn can be employed provide positive identification and quantification of individual glucosinolates. Such approaches offer sensitive tools for focused metabolomics analysis and screening of plant breeding lines. PMID- 18396303 TI - Species identification of human echinococcosis using histopathology and genotyping in northwestern China. AB - Human cystic echinococcosis, caused by infection with the larval stage of Echinococcus granulosus, and alveolar echinococcosis, caused by the larval form of E. multilocularis, are known to be important public health problems in western China. Echinococcus shiquicus is a new species of Echinococcus recently described in wildlife hosts from the eastern Tibetan plateau and its infectivity and/or pathogenicity in humans remain unknown. In the current study, parasite tissues from various organs were collected post-operatively from 68 echinococcosis patients from Sichuan and Qinghai provinces in eastern China. The tissues were examined by histopathology and genotyped using DNA sequencing and PCR-RFLP. Histopathologically, 38 human isolates were confirmed as E. granulosus and 30 as E. multilocularis. Mitochondrial cob gene sequencing and PCR-RFLP with rrnL as the target gene confirmed 33 of 53 of the isolates to have the G1 genotype of sheep/dog strain of E. granulosus as the only source of infection, while the remaining 20 isolates were identified as E. multilocularis. No infections were found to be caused by E. shiquicus. Additionally, 5 of 20 alveolar echinococcosis patients were confirmed to have intracranial metastases from primary hepatic alveolar echinococcosis lesions. All these cases originated from four provinces or autonomous regions but most were distributed in Sichuan and Qinghai provinces, where high prevalence rates of human alveolar echinococcosis and cystic echinococcosis were previously documented. PMID- 18396304 TI - Zinc inhibits ethanol-induced HepG2 cell apoptosis. AB - Alcohol consumption produces a variety of metabolic alterations in liver cells, associated with ethanol oxidation and with nonoxidative metabolism of ethanol, among others apoptosis of hepatocytes. As zinc is known as a potent antioxidant and an inhibitor of cell apoptosis, the aim of this paper was to investigate whether zinc supplementation could inhibit ethanol-induced HepG2 apoptosis, and whether this inhibition was connected with attenuation of oxidative stress and modulation of FasR/FasL system expression. The results indicated that zinc supplementation significantly inhibited ethanol-induced HepG2 cell apoptosis (measured by cytochrome c release from mitochondria and caspase-3 activation) by attenuation of reactive oxygen species (ROS) production, increase in the cellular level of GSH, inhibition of ethanol-induced sFasR and FasL overexpression and caspase-8 activation. These results indicate that zinc can inhibit ethanol induced hepatocyte apoptosis by several independent mechanisms, among others by an indirect antioxidative effect and probably by inhibition of caspase-8 and caspase-9 activation. PMID- 18396305 TI - Contribution of reactive oxygen species to para-aminophenol toxicity in LLC-PK1 cells. AB - para-aminophenol (PAP) causes nephrotoxicity by biochemical mechanisms that have not been fully elucidated. PAP can undergo enzymatic or non-enzymatic oxidation to form reactive intermediates. Using modulators of reactive oxygen species (ROS), the role of ROS in PAP toxicity in LLC-PK(1) cells was investigated. ROS formation was determined using a fluorescein derivative and viability using alamarBlue. Following treatment of cells with PAP, ROS formation occurred prior to loss of cell viability. Several modulators of ROS were used to identify the pathways involved in PAP toxicity. Viability was improved with catalase treatment, while viability was decreased when cells were treated with superoxide dismutase (SOD). Both catalase and SOD exert their effects outside of cells in the incubation medium, since there was no evidence of uptake of these enzymes in LLC-PK(1) cells. In cell-free incubations, hydrogen peroxide (H(2)O(2)) was produced when 0.5 mM PAP was included in the incubation medium. Further, SOD greatly increased and catalase greatly decreased H(2)O(2) production in these cell-free incubations. These data suggest that H(2)O(2) formed in the incubation medium contributes to loss of viability following PAP treatment. When cells were coincubated with 0.5 mM PAP and tiron, pyruvate, bathocuproine, 1, 10 phenanthroline, or dimethylthiourea (DMTU), ROS formation was decreased. However, there was minimal improvement in cell viability. Paradoxically, DMTU exacerbated PAP-induced loss of viability. These data suggest that ROS are generated in cells exposed to PAP but these species are not the predominant cause of cellular injury. PMID- 18396306 TI - SARS vaccine based on a replication-defective recombinant vesicular stomatitis virus is more potent than one based on a replication-competent vector. AB - A SARS vaccine based on a live-attenuated vesicular stomatitis virus (VSV) recombinant expressing the SARS-CoV S protein provides long-term protection of immunized mice from SARS-CoV infection (Kapadia, S.U., Rose, J. K., Lamirande, E., Vogel, L., Subbarao, K., Roberts, A., 2005. Long-term protection from SARS coronavirus infection conferred by a single immunization with an attenuated VSV based vaccine. Virology 340(2), 174-82.). Because it is difficult to obtain regulatory approval of vaccine based on live viruses, we constructed a replication-defective single-cycle VSV vector in which we replaced the VSV glycoprotein (G) gene with the SARS-CoV S gene. The virus was only able to infect cells when pseudotyped with the VSV G protein. We measured the effectiveness of immunization with the single-cycle vaccine in mice. We found that the vaccine given intramuscularly induced a neutralizing antibody response to SARS-CoV that was approximately ten-fold greater than that required for the protection from SARS-CoV infection, and significantly greater than that generated by the replication-competent vector expressing SARS-CoV S protein given by the same route. Our results, along with earlier studies showing potent induction of T-cell responses by single-cycle vectors, indicate that these vectors are excellent alternatives to live-attenuated VSV. PMID- 18396307 TI - Differential aging of motion processing mechanisms: evidence against general perceptual decline. AB - While the percentage of older people in our society is steadily increasing, knowledge about perceptual changes during healthy aging is still limited. We investigated age effects on visual motion perception in order to differentiate between general decline and specific vulnerabilities. A total of 119 subjects ranging in age from 20 to 82 years participated in our study. Perceptual thresholds for different types of motion information, including translational motion, expanding radial flow, and biological motion, were determined. Results revealed a substantial increase of thresholds for translational motion with age. Biological motion perception was only moderately affected by age. For both motion types, threshold elevation seemed to develop gradually with age. In contrast, we found stable radial flow analysis across lifespan. There was no evidence that age effects were dependent on gender. Results demonstrate that visual capabilities are not equally prone to age-related decline. Surprisingly, higher motion complexity might not be necessarily associated with more pronounced perceptual constraints. We suggest that differential age effects on the perception of specific motion types might indicate that specialized neuronal processing mechanisms differ in their vulnerability to physiological changes during aging. PMID- 18396309 TI - Complete treatment of olive pomace leachate by coagulation, activated-carbon adsorption and electrochemical oxidation. AB - A battery scheme comprising sequential alum coagulation, activated-carbon adsorption and electrochemical oxidation over boron-doped diamond electrodes to mineralize a leachate from olive pomace processing is demonstrated. The effect of coagulant and adsorbent concentration on treatment efficiency was assessed in the range 0.1-50 mM Al(3+) and 2.5-50 g/L activated-carbon and optimal conditions were established. Coagulation at 7.5mM Al(3+) resulted in substantial solids and color removal (i.e. 80% and 93%, respectively). This was accompanied by only 30% chemical oxygen demand (COD) reduction (initial COD was about 3,500 mg/L). The latter increased to 80% though when coagulation was coupled with adsorption at 25 g/L activated carbon. Electrochemical oxidation of the original effluent for 360 min led to 63% and 82% COD reduction at 10 and 20A current intensity, respectively. When this process was tested as a polishing stage following coagulation and adsorption, overall COD removal reached values of 92% and 97%, respectively. The final effluent was also colorless and solids free. However, the treated effluent still exhibited ecotoxicity possibly due to the formation of ecotoxic oxidation products. PMID- 18396308 TI - Personal names do not always survive the attentional blink: Behavioral evidence for a flexible locus of selection. AB - Models of the attentional blink phenomenon (AB) typically assume that unattended information is processed to the post-perceptual level prior to selection for access to consciousness. The present experiments test this assumption by manipulating the perceptual load of the first target task (T1) and whether the second target (T2) was the participant's own name or someone else's name. In three experiments, increasing T1-load increased the severity of the AB for personal names. The results suggest that selection during the AB is not fixed at the post-perceptual stage, but rather the stage at which selection occurs during the AB is flexible. PMID- 18396310 TI - Impact of shear force on the biofilm structure and performance of a membrane biofilm reactor for tertiary hydrogen-driven denitrification of municipal wastewater. AB - Hydrogen-driven denitrification using a hollow-fiber membrane biofilm reactor (MBfR) was evaluated for operation in tertiary wastewater treatment. Specific objectives were to evaluate the impact of different levels of shearing stress caused by mixing and nitrogen sparging on the biofilm structure and denitrification rates. Applying high shear force proved to be effective in improving denitrification rates by reducing the thickness of the biofilm. With intensive mixing a biofilm thickness of approximately 800 microm was maintained, while additional nitrogen sparging could further reduce the biofilm thickness to approximately 300 microm. The highest denitrification rates of 0.93 gN/m(2)d were obtained when biofilm thickness was lower than 500 microm. Lower extracellular polymeric substances (EPS) accumulation and carbohydrates to protein ratio observed in thinner biofilms allowed for higher nitrate removal in the system. No significant sloughing of biomass or change in total and soluble COD in the final effluent was observed under steady-state conditions. PMID- 18396311 TI - Organochlorine contaminants in hair of adolescents from Iassy, Romania. AB - Human hair samples (n=42) from Iassy county (Eastern Romania) collected in 2002 2003 from adolescents were analyzed for hexachlorocyclohexane (HCH) isomers, p,p' DDT and its metabolites, hexachlorobenzene (HCB), chlordane and metabolites and 5 polychlorinated biphenyl (PCB) congeners. Very low levels were found for HCB and oxychlordane (0.8 and 2.5 ng g(-1)) indicating a low usage of these pesticide formulations in the studied area. In case of HCHs, gamma-HCH isomer was measured at higher median concentrations (79 ng g(-1) hair) compared to the beta-HCH isomer (55 ng g(-1)), which generally is the most prevalent from HCHs. The DDTs profile consisted in p,p'-DDE and p,p'-DDT which levels correspond to 81% of sum DDTs. Very high median concentrations measured for p,p'-DDT (192 ng g(-1)) combined with lower values for p,p'-DDE/p,p'-DDT of 0.4 (from 0.20 to 2.0) in all hair samples suggest recent exposure to "fresh" DDT. When gender was considered, significantly higher concentrations for most of the investigated contaminants were found in girls hair compared to boys. Very high levels were found in the present study for HCHs and DDTs compared to samples from Greece, Western Europe and China. In case of PCBs, the levels found in Romanian samples were found to be in the same range compared to other previously published data. PMID- 18396312 TI - Effects of aging and sediment composition on hexachlorobenzene desorption resistance compared to oral bioavailability in rats. AB - Studies were conducted to assess the effects of black carbon, clay type and aging (1-1.5yr) on desorption and bioavailability of hexachlorobenzene (HCB) in spiked artificial sediments. Tenax (a super sorbent)-mediated desorption was used to examine the effects of these parameters on the physicochemical availability of HCB. The Tenax-mediated desorption of HCB from the four aged artificial sediments exhibited biphasic kinetics. The fast desorbing fractions ranged from 64.8% to 22.3%, showing reductions of 4.0-18.9% compared with freshly-spiked sediments. Statistical analysis on the fast desorbing fractions showed that all three treatment effects (i.e., montmorillonite clay, black carbon content, and aging) were significant. Two sediments with higher black carbon content exhibited much greater aging effects (i.e., greater reduction in fast desorbing fraction) than the other two sediments without the addition of black carbon. For both freshly spiked and aged sediments, the desorption resistant sediment-bound HCB (i.e., slow desorbing fraction) correlated reasonably well to previously reported rat fecal elimination of HCB, which is a measure of the non-bioavailable fraction of sediment-bound HCB. A similar correlation was also observed between fast desorbing fraction and previously reported accumulation of HCB in the rat body (carcass+skin). These observations suggest that physicochemical availability, as defined by the desorption of HCB from sediments, provides a reasonable prediction of the oral bioavailability of sediment-bound HCB to rats. These results showed that montmorillonite clay, black carbon and aging reduced physicochemical availability and ultimately bioavailability of sediment-bound HCB. PMID- 18396313 TI - Adsorption of arsenic(V) onto fly ash: a speciation-based approach. AB - Arsenic (As) poses a significant water quality problem and challenge for the environmental engineers and scientists in the world. The large volume of coal fly ash produced around the world is a potentially significant anthropogenic source of arsenic. Currently the leaching behavior of arsenic from fly ash is not well understood. Batch methods were used in this study to investigate arsenic leaching using a raw ash, and arsenic adsorption using a clean, washed ash. Experimental results indicated that pH had a significant effect on arsenic leaching or adsorption. Between pH 3 and 7, less arsenic was in the dissolved phase. When pH was less than 3 or greater than 7, increasing amounts of arsenic were leached or desorbed from fly ash. The leaching and adsorption behavior of arsenic was interpreted with the speciation of surface sites and arsenic. In a new approach, a speciation-based model was developed to quantify the arsenic adsorption as a function of pH and surface acidity parameters. This work is important in offering insight into the leaching mechanism of arsenic from coal fly ash, and providing a robust model based upon specific, measurable parameters to quantify arsenic adsorption by other solid media in addition to fly ash. PMID- 18396314 TI - Photocatalytic degradation of isoproturon herbicide over TiO2/Al-MCM-41 composite systems using solar light. AB - The present investigation covers immobilization of TiO2 using a simple solid state dispersion technique over mesoporous Al-MCM-41 support for the treatment of isoproturon herbicide. Catalysts are characterized by XRD, X-ray photo electron spectroscopy (XPS), surface area, UV-Vis diffused reflectance spectra (DRS), SEM and TEM. A detailed photocatalytic degradation study of isoproturon under solar light in aqueous suspensions is reported. The 10 wt% TiO2/Al-MCM-41 composite system found to be optimum with high degradation activity. The reaction follows pseudo-first order kinetics. The parameters like TiO2 loading over Al-MCM-41, amount of catalyst, concentration of substrate, pH effect, durability of the catalyst, activity comparison of TiO2 and Al-MCM-41 supported system are studied. The mineralization of isoproturon is monitored by TOC. Based on the degradation products detected through LC-MS, a plausible degradation mechanism is proposed. The data indicates that TiO2/Al-MCM-41 composite system is an effective photocatalyst for treatment of isoproturon in contaminated water. PMID- 18396315 TI - Simple digestion procedure followed by the azomethine-H method for accurate boron analysis and discrimination between its fractions in wastewater and soils. AB - Excess boron is a growing environmental problem. It often affects agricultural yields, where reuse of wastewater for irrigation is practiced. This problem raises the need for reliable, simple and economical methods to monitor boron concentrations in wastewater and soil extracts. One such method, the commonly used azomethine-H spectrophotometric method, suffers from color interference, originating from high concentrations of dissolved organic matter, when applied to many wastewater and agricultural soil extracts. Moreover, this method only quantifies free dissolved boron and lacks the ability to quantify boron that is adsorbed to either the dissolved organic matter or suspended solids that are present in the sample. This work suggests a modification of the standard azomethine-H method, in which the solution is digested with potassium persulfate prior to the standard procedure. We show that this pretreatment can overcome the color interference and lead to highly accurate and precise boron analyses in wastewater. In soil extracts, the boron concentrations obtained using the suggested procedure were better correlated to inductively coupled plasma (ICP) spectrometry results than those measured by the standard method, because whereas the standard method quantifies the free dissolved boron only, the modified method, like the ICP method, quantifies the total dissolved boron in the sample. Thus, the suggested modification can be used to quantify the respective distributions of free dissolved boron, boron adsorbed to dissolved organic matter and boron adsorbed to suspended solids in soil extracts and water samples. PMID- 18396316 TI - Growth-inhibitory effects of sulfonamides at different pH: dissimilar susceptibility patterns of a soil bacterium and a test bacterium used for antibiotic assays. AB - The ionic speciation of sulfonamides is pH-driven and this may be crucial for their bioavailability and sorption to soil constituents, as well as for their uptake into bacterial cells. The inhibition behaviour of a bacterial test strain (Pseudomonas aeruginosa; DSM 1117), which was grown in the presence of different concentrations of 8 sulfonamides at pH values from 5 to 8, could be predicted by models that take the speciation of sulfonamides in- and outside of bacterial cells into account. Assuming a pH of 7.5 inside the cells (pH homeostasis), the strongest inhibition was predicted for the lowest external pH and for sulfonamides with the lowest pK(a) values. Growth experiments with Ps. aeruginosa basically reflected this predicted behaviour. However, Pantoea agglomerans -- a bacterial strain isolated from arable soil -- behaved surprisingly different regarding its pH dependency: all sulfonamides showed the strongest effects at pH 7 to 8 instead of being most effective at lowest pH, although the pK(a) dependencies followed the same pattern. Experimental and modeling results could be brought into good agreement for P. agglomerans if the cell-internal pH was admitted to approximate the external pH instead of implying pH homeostasis for modeling calculations. Thus, besides the actual concentration of sulfonamides, the pH dependent mode of reaction of different bacteria to sulfonamides may additionally govern the population dynamics in soils. PMID- 18396317 TI - Constructed wetlands treating highway runoff in the central Mediterranean region. AB - Two free water surface (FWS) and two subsurface flow (SSF) pilot-size constructed wetlands treating highway runoff (HRO) were monitored over a period of two years (September 2005-August 2007). One FWS and one SSF were designed with a hydraulic retention time (HRT) of 12h, named FWS12 and SSF12, respectively, with each one capable of treating a maximum HRO of 12.6 m(3) d(-1). The other couple, named FWS24 and SSF24, respectively, was designed with an HRT of 24h, with each receiving a maximum HRO of 6.3 m(3) d(-1). The influent flowed from a highway section with a total surface 2752 m(2) on the island of Crete, Greece, in the heart of the South-Central Mediterranean region. Influent and effluent were monitored for COD, TSS, total N (TN), NO(3)(-) and total P (TP) concentrations. Furthermore, removal efficiencies were examined for heavy metals (Cu, Ni, Pb, Zn) for both years, while polycyclic aromatic compounds (PAHs) were examined for the period between September 2006 and August 2007. The influent had a two-year average COD value of 101 mg l(-1), whereas the mean values for TSS, TN, N-NO(3)( ) and TP were 203, 4.30, 1.25 and 4.17 mg l(-1), respectively. For Cu, Ni, Pb and Zn the respective two-year mean influent concentrations were 56, 114, 49 and 250 microg l(-1). Mean concentration of total PAHs in runoff (summation operator PAHs, 16 compounds) were 12.01 microg l(-1). The performance among the four beds was not significantly different according to ANOVA analysis followed by Tukey test (at p<0.05) for almost all the above physicochemical parameters, suggesting that all systems performed in a similar way. All studied systems, achieved a mean of two-year removal efficiencies of 47% for COD, 89% for TSS, 49% for TN, 58% for N-NO(3)(-), 60% for TP, 47% for Cu, 23% for Ni, 33% for Pb, 61% for Zn and 59% for summation operator PAHs (16 compounds). PMID- 18396318 TI - Isotopic dietary analysis of a Neanderthal and associated fauna from the site of Jonzac (Charente-Maritime), France. AB - We report here on the isotopic analysis (carbon and nitrogen) of collagen extracted from a Neanderthal tooth and animal bone from the late Mousterian site of Jonzac (Charente-Maritime, France). This study was undertaken to test whether the isotopic evidence indicates that animal protein was the main source of dietary protein for this relatively late Neanderthal, as suggested by previous studies. This was of particular interest here because this is the first isotopic study of a relatively late Neanderthal associated with Mousterian of Acheulian Tradition (MTA, dating to approximately 55,000 to 40,000 BP) technology. We found that the Jonzac Neanderthal had isotopic values consistent with a diet in which the main protein sources were large herbivores, particularly bovids and horses. We also found evidence of different dietary niches between the Neanderthal and a hyena at the site, with the hyena consuming mainly reindeer. PMID- 18396319 TI - Organochlorine pesticides in ambient air in Durban, South Africa. AB - Despite the existence of numerous sources and continuing use, information regarding emissions and airborne concentrations of organochlorine pesticides in Africa is extremely limited. This paper presents results of a monitoring program conducted in Durban, South Africa that was designed to characterize levels, trends and possible sources of pesticides in both industrial and residential areas. Three monitoring sites were established, two in an industrialized area in the southern part of the city, and the third in a northern residential area. Particulate and vapor samples were sampled over the 2004-5 period and analyzed by GC/MS to estimate long-term levels of a wide range of pesticides. Based on a year of sampling, the sites had comparable levels of many pesticides with exceptions of alpha-chlordane and lindane. Levels of p,p'-DDT (42+/-27 pg m(-3)) and its derivatives were relatively high and showed an unusual mixture with high levels of p,p'-DDD (12+/-11 pg m(-3)). Other pesticides detected and quantified included aldrin, chlordanes, hexachlorobenzene and dieldrin. Potential source areas, identified using concentration patterns, local and regional gradients, compositional information and trajectory analyses, suggest that chlordane and lindane arise from both local sources as well as regional/global sources; DDT from regional sources elsewhere in South Africa, Africa and India; and most of the other long-lived pesticides detected, including gamma-nonachlor, hexachlorobenzene and toxaphene, from global sources. This monitoring results, which represent the most detailed study to date of pesticides in air in Africa, serve several purposes, including documenting the presence and use of long-banned pesticides like aldrin, aiding the understanding of the fate of persistent compounds, identifying pollutants that may contribute to health problems, and informing decision-making aimed at reducing exposures and risks. PMID- 18396320 TI - Atmospheric mercury depletion event study in Ny-Alesund (Svalbard) in spring 2005. Deposition and transformation of Hg in surface snow during springtime. AB - A field campaign was conducted in Ny-Alesund (78 degrees 54'N, 11 degrees 53'E), Svalbard (Norway) during April and May 2005. An Atmospheric Mercury (Hg) Depletion Event (AMDE) was observed from the morning of April 24 until the evening of April 27. Transport of already Hg and ozone (O3) depleted air masses could explain this observed depletion. Due to a snowfall event during the AMDE, surface snow Hg concentrations increased two fold. Hg deposition took place over a short period of time corresponding to 3-4 days. More than 80% of the deposited Hg was estimated to be reemitted back to the atmosphere in the days following the event. During the campaign, we observed night and day variations in surface snow Hg concentrations, which may be the result of gaseous elemental mercury (GEM) oxidation to divalent Hg at the snow/air interface by daylight surface snow chemistry. Finally, a decrease in the reactive Hg (HgR) fraction of total Hg (HgT) in the surface snow was observed during spring. We postulate that the transformation of HgR to a more stable form may occur in Arctic snow during spring. PMID- 18396321 TI - Pharmaceutical compounds in the wastewater process stream in Northwest Ohio. AB - In order to add to the current state of knowledge regarding occurrence and fate of Pharmaceutical and Personal Care Products (PPCP's) in the environment, influent, effluent and biosolids from three wastewater treatment facilities in Northwest Ohio, USA, and a stream containing effluent discharge from a rural treatment facility were analyzed. The three WWTP facilities vary in size and in community served, but are all Class B facilities. One facility was sampled multiple times in order to assess temporal variability. Twenty compounds including several classes of antibiotics, acidic pharmaceuticals, and prescribed medications were analyzed using ultrasonication extraction, SPE cleanup and liquid chromatography-electrospray ionization tandem mass spectrometry. The highest number of compounds and the greatest concentrations were found in the influent from the largest and most industrial WWTP facility. Short-term temporal variability was minimal at this facility. Many compounds, such as clarithromycin, salicylic acid and gemfibrizol were found at concentrations more than one order of magnitude higher than found in the effluent samples. Effluent waters contained elevated levels of carbamazepine, clindamycin and sulfamethoxazole. Differences in composition and concentration of effluent waters between facilities existed. Biosolid samples from two different facilities were very similar in PPCP composition, although concentrations varied. Ciprofloxacin was found in biosolids at concentrations (up to 46 mug/kg dry mass) lower than values reported elsewhere. Diclofenac survived the WWTP process and was found to persist in stream water incorporating effluent discharge. The low variability within one plant, as compared to the variability found among different wastewater treatment plants locally and in the literature is likely due to differences in population, PPCP usage, plant operations and/or local environment. These data are presented here for comparison with this emerging set of environmental compounds of concern. PMID- 18396322 TI - Transverse colectomy in ovarian cancer surgical cytoreduction: operative technique and clinical outcome. AB - OBJECTIVE: To describe the operative techniques and associated clinical outcomes of patients undergoing transverse colectomy as a component of cytoreductive surgery for advanced or recurrent ovarian cancer. METHODS: Thirty-nine patients underwent transverse colectomy as part of primary (n=33) or secondary (n=6) cytoreductive surgery for ovarian cancer between 1/97 and 4/07. The surgical techniques, associated morbidity, and clinical outcomes are described. RESULTS: Among primary surgery patients, 75.6% had Stage IIIC disease, and 24.2% had Stage IV disease. Transverse colon surgery consisted of: partial colectomy in 33 cases and total transverse colectomy in 6 cases. Transverse colectomy with rectosigmoid colectomy was performed in 61.5% of patients, with two separate colonic anastomoses in 48.7%. The majority (89.7%) of transverse colon anastomoses were stapled, most commonly a functional end-to-end colocolostomy. Two patients required end colostomy. The median EBL was 500 cm(3). Residual disease was: no gross in 33.3%, 0.1-1.0 cm in 59.0%, and >1 cm in 7.7% of patients. Post operative morbidity occurred in 25.6% of patients, with a fistula rate of 5.1% and a mortality rate of 2.6%. The median survival time after primary surgery was 68.3 months. CONCLUSIONS: Transverse colectomy can contribute significantly to a maximal ovarian cancer cytoreductive surgical effort and carries acceptable morbidity. Resection of a non-contiguous segment of rectosigmoid colon is frequently necessary, and placement of two separate colonic anastomoses is associated with a low risk of anastomotic breakdown. PMID- 18396323 TI - Filaggrin mutations, atopic eczema, hay fever, and asthma in children. AB - BACKGROUND: Mutations in the filaggrin gene (FLG) have been shown to play a significant role in ichthyosis vulgaris and eczema, 2 common chronic skin diseases. However, their role in the development of other atopic diseases such as asthma and rhinitis has not yet been clarified in large population-based studies. OBJECTIVES: To study the effect of FLG mutations at the population level and their effect on other atopic phenotypes. METHODS: Association analysis of the 2 common FLG-null mutations R501X and 2282del4 and 3 recently identified rare FLG variants (R2447X, S3247X, 3702delG) was performed on our cross-sectional population of German children (n = 3099) recruited as part of the International Study of Asthma and Allergies in Childhood II in Munich (n = 1159) and Dresden (n = 1940). RESULTS: FLG variants increased the risk for eczema more than 3-fold (odds ratio [OR], 3.12; 95% CI, 2.33-4.173; P = 2.5 x 10(-14); population attributable risk, 13.5%). Independent of eczema, FLG mutations conferred a substantial risk for allergic rhinitis (OR, 2.64; 95% CI, 1.76-4.00; P = 2.5 x 10(-6); population-attributable risk, 10.8%). Nasal biopsies demonstrated strong filaggrin expression in the cornified epithelium of the nasal vestibular lining, but not the transitional and respiratory nasal epithelia. In contrast, the association with asthma (OR, 1.79; 95% CI, 1.19-2.68; P = .0048) was restricted to asthma occurring in the context of eczema, and there was a strong association with the complex phenotype eczema plus asthma (OR, 3.49; 95% CI, 2.00-6.08; P = 1.0 x 10(-5)). CONCLUSION: Our results suggest that FLG mutations are key organ specific factors predominantly affecting the development of eczema and confer significant risks of allergic sensitization and allergic rhinitis as well as asthma in the context of eczema. PMID- 18396324 TI - Effects of naphthalene and phenanthrene on visual and prey capture endpoints during early stages of the dourado Salminus brasiliensis. AB - Naphthalene (NAP) and phenanthrene (PHE) effects on Salminus brasiliensis, a carnivorous freshwater fish, were investigated using behavioral tests. Larval stages of S. brasiliensis were exposed to water concentrations of 0, 0.04 mg/l, 0.20mg/l and 0.50mg/l for naphthalene and 0, 0.01 mg/l, 0.05 mg/l and 0.1mg/l for phenanthrene during two developmental phases. The prey fish Prochilodus lineatus were not exposed. Visual acuity was measured at the end of phase 2 in individual S. brasiliensis, which were also tested at the end of each phase for number of attacks on prey, number of prey captured, prey capture efficiency, and distance swam. Vision was impaired by PHE exposure, as acuity angles increased in exposed fish. At Stage I 2.3+/-1.2 prey were captured with 46% efficiency in controls compared to 0.4+/-0.3 prey captured with 13.4% efficiency in fish exposed to 0.05 mg PHEl(-1), the LOEL for both endpoints. At Stage II 4.0+/-1.1 preys were captured in controls compared to 2.5+/-0.9 preys captured in fish exposed to 0.01 mg PHEl(-1), the LOEL. Stage II control fish captured prey with 70% efficiency compared to 30% efficiency at Stage II fish exposed to 0.05 mg PHEl(-1), the LOEL. Distance swam was not affected by either NAP or PHE exposure. The exposure of larval stages of S. brasiliensis to realistic water concentrations of PHE impairs foraging skills and could affect recruitment of the species. PMID- 18396325 TI - Teasing apart activities of different types of ABC efflux pumps in bivalve gills using the concepts of independent action and concentration addition. AB - Fluorescent dyes and inhibitor compounds are commonly used to detect activity of multixenobiotic resistance (MXR) efflux pumps in marine invertebrates. We here address the question whether compounds acting as specific inhibitors of certain mammalian transporters can be used in dye efflux assays to distinguish different transporter activities in gill tissue from a marine mussel. We quantified effects of PSC833, a specific inhibitor of mammalian P-gp (P-glycoprotein, ABCB1), and MK571, which blocks MRP (Multidrug resistance associated protein, ABCC) type transporters, on calcein-am efflux in gill tissue of Mytilus californianus. Calcein-am acts as a substrate of both P-gp and MRP. Effects of single compounds and mixtures were determined and combined effect models predicting independent action (IA) and concentration addition (CA) of the chemicals were applied. Effect values predicted by IA showed better correspondence with the experimentally obtained data. This indicates that the inhibitor compounds target different mechanisms of calcein-am efflux and points to P-gp and MRP activities in mussel gills. Our approach could be a simple way for identifying the efflux transporter types targeted by chemosensitizers, including environmentally relevant compounds, in native tissues from marine invertebrates. PMID- 18396326 TI - Effects of oxidative stress on protein thiols and disulphides in Mytilus edulis revealed by proteomics: actin and protein disulphide isomerase are redox targets. AB - Many proteins contain cysteines which are sensitive to oxidation. This is sometimes reversible through interaction with glutathione, glutaredoxin or thioredoxin systems making these proteins potential sensors of oxidative stress. In this study we analysed whether there was an increase in mixed disulphide bond (-S-S-) formation in the blue mussel Mytilus edulis in response to menadione. This was achieved by initially blocking reduced thiols with N-ethylmaleimide, -S S- were then reduced with dithiothreitol (DTT) and labelled with 5 iodoacetamidofluorescein (5-IAF). Free -SHs were also labelled directly with 5 IAF. Separations were performed on 1D or 2D SDS PAGE and images analysed. There was an increase in -S-S- in response to menadione and detection of changes in oxidised proteins was easier than that of changes in the amount of reduced proteins. Protein disulphide isomerase (PDI) was labelled both as -SH and -S-S-, underlining its involvement in the redox status of the animal. A glutathione transferase (GST P1-1) forms an inter-chain disulphide bridge in response to menadione. PMID- 18396327 TI - Use of immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as new "in vitro" tool to investigate effects of environmental contaminants. AB - The aim of the present study was to propose the immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as a new "in vitro" tool to explore the susceptibility of these marine mammals to different xenobiotic compounds. The cell lines were cultured from integument biopsies of free-ranging and stranded cetaceans (dead within 12h). Using the indirect immunofluorescence assay, we detected endogenous proteins induced by different contaminants. Here we present the method used for qualitative and quantitative evaluation of cytochromes P450 (CYP1A1 and CYP2B) induced by some POPs (DDTs and PCBs) and emerging contaminants (PBDEs) in fibroblast cell cultures of striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Immunofluorescence was quantified with a specially designed Olympus macro, DetectIntZ. A major result was the possibility of using this "in vitro" assay to quantify induction of endogenous proteins. PMID- 18396328 TI - Intersexuality in crustaceans: genetic, individual and population level effects. AB - Scientists investigating toxicants such as endocrine disrupting chemicals (EDCs) at the cellular at the sub-cellular level are often faced with criticisms as to how these effects can be extrapolated to the level of individuals and their populations. This report aims to provide an overview of the studies undertaken on crustacean model, Echinogammarus marinus LEACH (AMPHIPODA), and intersex phenotypes, at the individual and population levels, and provide additional emergent data at the genomic level. These, normal and intersex, males and females have been investigated by cross-hybridisation microarray analysis and specific sexually dimorphic genes and corresponding properties identified between each sexual phenotype. The morphology, physiology and histology of these intersexes have been investigated in detail and a number of reproductive costs have been identified including reduced fecundity and fertility. These costs have been incorporated into a population model and simulated over a ten-year period to ascertain how different levels of intersexuality affect the stability of populations. Based on the information gained through study of intersex models (with known endocrine dysfunction) together with the substantial quantity of historical data relating to effects of chemicals on amphipod fecundity, growth and mortality, the development of appropriate biomarkers is nearer to being assessed from the level of genes to that of the population. PMID- 18396329 TI - The effect of sustained delivery of vascular endothelial growth factor on angiogenesis in tissue-engineered intestine. AB - Our group has previously created a functional neointestine that is capable of restoring absorptive function. However, the endogenous level of vascular endothelial growth factor (VEGF) is markedly reduced in the construct compared to native bowel. Therefore, we wanted to locally deliver VEGF in a sustained fashion to upregulate angiogenesis in the neointestine. Rat recombinant VEGF was encapsulated in poly(lactide-co-glycolide) microspheres by a double emulsion method. Release kinetics and bioactivity were determined in vitro. Tissue engineered intestine was generated by seeding donor neonatal rat intestinal organoid units onto a biodegradable polyglycolic acid scaffold along with VEGF containing or empty microspheres, and wrapped in the omentum of recipient rats. After 4 weeks, the neointestinal cysts were analyzed for morphometry, VEGF levels, epithelial proliferation, and capillary density. Sustained release of biologically active VEGF was confirmed by in vitro studies. Intestinal constructs with VEGF microspheres were significantly larger than those containing empty microspheres. Tissue VEGF levels were significantly higher in neointestine loaded with encapsulated VEGF compared to those without growth factor. Epithelial cellular proliferation and capillary density were significantly increased in the VEGF-containing neointestinal constructs compared to empty constructs. Tissue engineered intestine responds to sustained delivery of VEGF by upregulating microvasculature and epithelial proliferation. PMID- 18396330 TI - Preparation and characterization of pH- and temperature-sensitive pullulan microspheres for controlled release of drugs. AB - Most part of pH- and temperature-sensitive microspheres used for the controlled delivery of drugs are not biodegradable. Therefore, the aim of this work is to prepare pH- and temperature-sensitive microspheres from biodegradable and biocompatible natural polymers. Pullulan microspheres were prepared by suspension cross-linking with epichlorohydrin of an aqueous solution of the polymer. In order to confer them temperature sensitivity, poly(N-isopropylacrylamide-co acrylamide) was grafted onto pullulan microspheres. Then, the pH-sensitive units (-COOH) were introduced by reaction between the remaining -OH groups of the pullulan with succinic anhydride. The grafted pullulan microspheres are more hydrophilic than pullulan microspheres, their swelling degree as well as water regain increase significantly. The thermo-sensitivity of the carboxylated microspheres depends to the number and the ionization form (-COOH/-COO(-)) of carboxylic groups. At a low exchange capacity (0.35 meq/g), microspheres are thermo-sensitive both in the protonated and deprotonated form of -COOH groups. At a higher exchange capacity (2.25 meq/g), microspheres are almost unswellable in the protonated form and swell extensively in the ionized form (up to 28 times than their dried form) loosing in a great extent the thermo-sensitive properties. In isotonic phosphate buffer pH=7.4, both thermo-sensitive and pH/thermo sensitive microspheres possess a phase transition temperature close to that of the human body temperature. Loading and release profiles of lysozyme, taken as a molecular model system, were investigated. PMID- 18396331 TI - Three-dimensional extracellular matrix-directed cardioprogenitor differentiation: systematic modulation of a synthetic cell-responsive PEG-hydrogel. AB - We show that synthetic three-dimensional (3D) matrix metalloproteinase (MMP) sensitive poly(ethylene glycol) (PEG)-based hydrogels can direct differentiation of pluripotent cardioprogenitors, using P19 embryonal carcinoma (EC) cells as a model, along a cardiac lineage in vitro. In order to systematically probe 3D matrix effects on P19 EC differentiation, matrix elasticity, MMP-sensitivity and the concentration of a matrix-bound RGDSP peptide were modulated. Soft matrices (E=322+/-64.2 Pa, stoichiometric ratio: 0.8), mimicking the elasticity of embryonic cardiac tissue, increased the fraction of cells expressing the early cardiac transcription factor Nkx2.5 around 2-fold compared to embryoid bodies (EB) in suspension. In contrast, stiffer matrices (E=4,036+/-419.6 Pa, stoichiometric ratio: 1.2) decreased the number of Nkx2.5-positive cells significantly. Further indicators of cardiac maturation were promoted by ligation of integrins relevant in early cardiac development (alpha(5)beta(1,) alpha(v)beta(3)) by the RGDSP ligand in combination with the MMP-sensitivity of the matrix, with a 6-fold increased amount of myosin heavy chain (MHC)-positive cells as compared to EB in suspension. This precisely controlled 3D culture system thus may serve as a potential alternative to natural matrices for engineering cardiac tissue structures for cell culture and potentially therapeutic applications. PMID- 18396332 TI - Using thermal energy produced by irradiation of Mn-Zn ferrite magnetic nanoparticles (MZF-NPs) for heat-inducible gene expression. AB - One of the main advantages of gene therapy over traditional therapy is the potential to target the expression of therapeutic genes in desired cells or tissues. To achieve targeted gene expression, we developed a novel heat-inducible gene expression system in which thermal energy generated by Mn-Zn ferrite magnetic nanoparticles (MZF-NPs) under an alternating magnetic field (AMF) was used to activate gene expression. MZF-NPs, obtained by co-precipitation method, were firstly surface modified with cation poly(ethylenimine) (PEI). Then thermodynamic test of various doses of MZF-NPs was preformed in vivo and in vitro. PEI-MZF-NPs showed good DNA binding ability and high transfection efficiency. In AMF, they could rise to a steady temperature. To analyze the heat induced gene expression under an AMF, we combined P1730OR vector transfection with hyperthermia produced by irradiation of MZF-NPs. By using LacZ gene as a reporter gene and Hsp70 as a promoter, it was demonstrated that expression of a heterogeneous gene could be elevated to 10 to 500-fold over background by moderate hyperthermia (added 12.24 or 25.81 mg MZF-NPs to growth medium) in tissue cultured cells. When injected with 2.6 or 4.6 mg MZF-NPs, the temperature of tumor-bearing nude mice could rise to 39.5 or 42.8 degrees C, respectively, and the beta-gal concentration could increase up to 3.8 or 8.1 mU/mg proteins accordingly 1 day after hyperthermia treatment. Our results therefore supported hyperthermia produced by irradiation of MZF-NPs under an AMF as a feasible approach for targeted heat-induced gene expression. This novel system made use of the relative low Curie point of MZF-NPs to control the in vivo hyperthermia temperature and therefore acquired safe and effective heat-inducible transgene expression. PMID- 18396333 TI - Targeted delivery of paclitaxel using folate-decorated poly(lactide)-vitamin E TPGS nanoparticles. AB - We synthesized nanoparticles (NPs) of the blend of two-component copolymers for targeted chemotherapy with paclitaxel used as model drug. One component is poly(lactide)-D-alpha-tocopheryl polyethylene glycol succinate (PLA-TPGS), which is of desired hydrophobic-lipophilic balance, and another is TPGS-COOH, which facilitates the folate conjugation for targeting. The nanoparticles of the two copolymer blend at various component ratio were prepared by the solvent extraction/evaporation single emulsion method and then decorated by folate, which were characterized by laser light scattering (LLS) for particles' size and size distribution, zeta potential analyzer for surface charge, and X-ray photoelectron spectroscopy (XPS) for surface chemistry. The drug encapsulation efficiency (EE) and in vitro drug release were measured by high performance liquid chromatography (HPLC). The targeting effect was investigated in vitro by cancer cell uptake of coumarin-6-loaded NPs and further confirmed by cytotoxicity of cancer cells treated with the drug formulated in the NPs. We showed that the NP formulation has great advantages vs the pristine drug in achieving better therapeutic effect, which increased 8.68% for MCF-7 breast cancer cells, and that the folate decoration can significantly promote targeted delivery of the drug into the corresponding cancer cells and thus enhance its therapeutic effect, which increased 24.4% for the NP formulation of 16.7% TPGS-COOH component and 31.1% for the NP formulation of 33.3% TPGS-COOH component after 24h treatment at the same 25 microg/ml paclitaxel concentration. The experiments on C6 glioma cells further confirmed these advantages. PMID- 18396334 TI - Optimized HPLC method for tramadol and O-desmethyl tramadol determination in human plasma. AB - The optimized method for HPLC determination of tramadol and its metabolite O desmethyl tramadol in human plasma using sotalol as internal standard has been developed and validated by a new approach. The determination by fluorescence detection was performed on re-eluted solution, obtained after liquid-liquid extraction with ethyl acetate of the three analytes from plasma. The chromatographic separation of tramadol under a gradient elution was achieved at a temperature of 15 degrees C with a RP-18 column, guarded by a C18 precolumn. The mobile phase was a mixed aqueous solution containing ortho-phosphoric acid, triethylamine, acetonitrile and methanol in a complex gradient mode. The quantitative determination of tramadol was performed at different successive pairs of excitation/emission wavelengths (200/300 nm, 200/295 nm, 212/305 nm) with lower limits of quantification: LLOQ=4.078 ng/ml for tramadol, respectively LLOQ=3.271 ng/ml for O-desmethyl tramadol. For the LLOQ limits, were calculated the values of the coefficient of variation and difference between mean and the nominal concentration. For tramadol analyte they were CV%=5.147% and bias%= 7.273% in the intra-days and CV%=4.894% and bias%=0.836% in the between-days assay, respectively for the metabolite O-desmethyl tramadol they were CV%=11.517% and bias%=0.337% in the intra-days and CV%=6.41% and bias%=3.259% in the between days assay. In addition, the stabilities of the analytes were verified in different conditions. Both, tramadol and its metabolite proved to be stable in plasma for four weeks, frozen at -20 degrees C, but also for 48 h at 15 degrees C in the re-eluted solution after liquid-liquid extraction. PMID- 18396335 TI - Nitric oxide production of T lymphocytes is increased in rheumatoid arthritis. AB - Experimental and clinical evidence for T cell involvement in the pathology of rheumatoid arthritis (RA) is compelling, and points to a local dysregulation of T cell function in the inflamed joint. Nitric oxide (NO) has been shown to regulate T cell function under physiological conditions, but overproduction of NO may contribute to lymphocyte dysfunction characteristic of RA. Several investigations in patients with RA have documented evidence of increased NO synthesis, but these studies have focused largely on macrophage-derived NO and its impact on innate immune and inflammatory responses. In this study, we set out to explore the contribution that T cells make to NO production. We find that T cells from RA patients produce >2.5 times more NO than healthy donor T cells (p<0.001). Although NO is an important physiological mediator of mitochondrial biogenesis, mitochondrial mass is similar in RA and control T cells. In contrast, increased NO production is associated with increased cytoplasmic Ca(2+) concentrations in RA T cells (p<0.001). In vitro treatment of human peripheral blood lymphocytes, or Jurkat cells with TNF increases NO production (p=0.006 and p=0.001, respectively), whilst infliximab treatment in RA patients decreases T cell derived NO production within 6 weeks of the first infusion (p=0.005). Together, these data indicate that TNF induced NO production in T lymphocytes may contribute to perturbations of immune homeostasis in RA. PMID- 18396336 TI - Role of Erk1/2 activation in prion disease pathogenesis: absence of CCR1 leads to increased Erk1/2 activation and accelerated disease progression. AB - Prion diseases are neurodegenerative infections with gliosis and vacuolation. The mechanisms of degeneration remain unclear, but chemokines may be important. In current experiments CCR1 knock-out (KO) mice succumbed more rapidly to scrapie infection than WT controls. Infected KO mice had upregulation of CCL3, a CCR1 ligand, and CCR5, a receptor with specificity for CCL3. Both infected KO and WT mice had upregulation of CCR5-mediated signaling involving activation of Erk1/2 in astrocytes; however, activation was earlier in KO mice suggesting a role in pathogenesis. In both mouse strains activation of the Erk1/2 pathway may lead to astrocyte dysfunction resulting in neurodegeneration. PMID- 18396337 TI - Relapses in multiple sclerosis are associated with increased CD8+ T-cell mediated cytotoxicity in CSF. AB - MS is thought to be mediated by CD4(+) T-helper cells. To investigate the importance of CD8(+) cytotoxic T-cells in MS we analyzed peripheral blood T-cells by DNA microarray, and plasma and CSF levels of granzymes from MS patients and controls. Cytotoxic gene expression was decreased in peripheral T-cells from RRMS patients whereas plasma levels of granzymes were unchanged. However, granzyme levels were elevated in the CSF of RRMS patients at relapse compared with controls and remission. Thus, CD8+ T-cell-mediated cytotoxicity is confined to the CSF/CNS compartment in RRMS patients and may be involved in the immunopathogenesis of clinical relapses. PMID- 18396338 TI - Chronic exposure of human macrophages in vitro to morphine and methadone induces a putative tolerant/dependent state. AB - We have shown previously that whereas acute exposure of cultured murine peritoneal macrophages inhibits phagocytosis, chronic exposure results in a putative tolerant/dependent state. We now report similar observations using human cultured monocyte-derived macrophages (hMDM) from a control population and from methadone patients. With hMDM, acute exposure to morphine and methadone inhibited phagocytosis in a dose-dependent manner. In contrast, chronic exposure resulted in eventual normalization of phagocytosis, indicating that a putative tolerant state to the opiates had developed. When opiates were withdrawn from chronically exposed, tolerized hMDM, phagocytosis was once again depressed. The duration of withdrawal-induced depression lasted several hours, which is much longer than evidenced previously with murine macrophages. These data identify well with various in vivo studies on immune effects of opiate withdrawal; and, in so-doing, supplement ongoing speculation that opiate withdrawal is likely to have serious impact on host defenses of street heroin addicts. PMID- 18396339 TI - Learning and memory in 5-HT(1A)-receptor mutant mice. AB - The serotonin 1A (5-HT(1A))-receptor is involved in a wide range of physiological functions, but has also been implicated in the pathophysiology of anxiety disorders and depression. Although the 5-HT(1A)-receptor is one of the best described receptor subtypes of the serotonergic system, its complex distribution pattern, pre- and postsynaptic localisation, and its impact on various neurotransmitters aggravate the attribution of 5-HT(1A)-agonistic effects to behavioural outcomes. The role of 5-HT(1A)-receptors for cognitive processes seems undisputed. However, the exact involvement of pre- and postsynaptic sites remains unexplained. Genetically modified animals are a complementary approach to pharmacological studies for further investigations of the role of the 5-HT(1A) receptor. Next to 5-HT(1A)-receptor knockout mice, two transgenic mouse lines exist that either overexpress the 5-HT(1A)-receptor transiently or permanently. The latter mouse line stands out due to the fact that a distinct overexpression is primarily found in the outer cortical layers and hippocampus, both projection areas of serotonergic neurons. Here, we discuss the findings obtained from 5 HT(1A)-receptor knockout and overexpressing mice concerning their learning and memory abilities. PMID- 18396340 TI - Effects of mid-jejunal compared to duodenal glucose infusion on peptide hormone release and appetite in healthy men. AB - INTRODUCTION: Cells containing GIP and CCK predominate in the upper small intestine, while those containing GLP-1 are located more distally. Our aim was to compare the hormonal, glycemic and appetite responses to different sites of glucose delivery. METHODS: Ten healthy males were each studied twice, in randomized order. A catheter was positioned with openings 15 cm beyond the pylorus ("duodenal"), and 100 cm beyond ("mid-jejunal"). On one day, glucose was infused into the duodenum (1 kcal/min) and saline into the mid-jejunum, for 90 min. On the other day, the infusion sites were reversed. Blood was sampled frequently, and hunger was scored by questionnaires. The tube was removed and energy intake measured from a buffet meal. RESULTS: Stimulation of CCK and suppression of hunger were greater (each P<0.05), and energy intake less (P=0.05), with duodenal compared to mid-jejunal glucose infusion. Blood glucose, GIP, and insulin did not differ, and there was minimal GLP-1 increment on either day. CONCLUSIONS: There is regional variation in CCK, but not incretin hormone release, in the upper small intestine, and modest differences in the site of glucose exposure affect appetite and energy intake. PMID- 18396341 TI - Long-term incidence of atrial fibrillation and flutter after transcatheter atrial septal defect closure in adults. AB - BACKGROUND: The long-term risk of atrial fibrillation and flutter (AFF) after trascatheter atrial septal defects (ASD) closure in adults is unknown. METHODS: We studied 134 patients who have undergone transcatheter ASD closure at our institution at an age of > 18 years (mean age 39 +/- 16 years); Patients were followed-up for 4.8 +/- 2.7 years (range 0.8-9.6 years). We assessed the presence of AFF both before and after ASD closure using standard 12 lead ECGs or 24h ambulatory Holter monitors. RESULTS: 13 patients (10%) had documented AFF before the procedure (paroxysmal in 6, permanent in 7). Patients with AFF before the procedure were older (p < 0.0001), and had worse clinical condition (p = 0.0008). Patients without a history of AFF before the procedure and those who experienced paroxysmal AFF before the procedure had a very low annual risk (0%) of subsequent permanent AFF at long-term follow-up. Four patients with permanent AFF before the procedure (onset of < 12months) underwent electrical cardioversion immediately before ASD closure. Two of them (50%) are in sinus rhythm after 4.1 and 7.0 years, respectively. CONCLUSIONS: Transcatheter ASD closure performed in adults with no history of AFF or with a history of paroxysmal AFF before closure seems to protect from development of AFF in the long-term. In selected patients with permanent AFF at closure, device ASD closure together with arrhythmia cardioversion might be able to restore and maintain sinus rhythm in the long term. PMID- 18396342 TI - Clip-based arterial haemostasis after antegrade common femoral artery puncture. AB - BACKGROUND: Given the frequent involvement of infra-popliteal arteries, an ipsilateral antegrade common femoral artery puncture (ACFAP) is usually preferred to a contralateral retrograde femoral access for percutaneous transluminal angioplasty (PTA) in patients with critical limb ischemia (CLI). Because of the frequent difficulty to get a sufficient manual pressure on the puncture site, ACFAP is burdened by a high number of bleeding local complications, including retroperitoneal haematoma. We report a series of patients who consecutively received a clip-based arterial closure device after ACFAP and ipsilateral PTA for CLI. METHODS: Thirty patients (73+/-6 years; 18 men; 100% diabetes) admitted to our hospital because of CLI consecutively underwent peripheral PTA after an ACFAP and received a clip-based arterial closure device. Time to haemostasis was defined as the interval elapsed between clip deployment and first observed haemostasis. All patients were mobilized after 6 h. Follow-up was 30 days. RESULTS: All patients were on double anti-platelet therapy. At the end of the procedure, Activation Clotting Time was 226+/-37 s. Procedural success in delivering the clip was 100%. Time to haemostasis was 21+/-19 s. No major local vascular complications and in particular no retroperitoneal bleeding were documented. All patients could be discharged within the following 3 days. No major complications were noted during a 30 days follow-up. CONCLUSIONS: The use of a clip-based arterial closure device after ACFAP for peripheral PTA in CLI seems to be safe and effective. PMID- 18396343 TI - Impulsivity, anxiety, and individual differences in evoked and induced brain oscillations. AB - There is much evidence to confirm the view that slow waves of delta and theta ranges are linked to activation whereas alpha oscillations are somehow related to inhibition processes. In the domain of individual differences, impulsive behavior is frequently associated with increased spectral power of slow oscillations whereas Behavioral Inhibition (BI) may be associated with higher reactivity within alpha band. Here it is hypothesized that Impulsivity would predispose to higher responding in low frequencies whereas BI would predispose to higher responding within alpha band. In a sample of 51 subjects, evoked and induced responses to auditory stimuli were studied in two experimental sessions: 1) in a simple discrimination task and 2) in a stop-signal task in which subjects had to be ready to withdraw the prepared motor response. Impulsive subjects showed higher baseline delta, theta and alpha power and higher magnitude of induced responses in low frequencies. They also showed lower phase-locking in low frequencies to auditory stimuli and higher phase-locking to the overt behavioral response onset. High BI scorers showed higher baseline alpha power and higher desynchronization in this frequency band in response to stimuli. The reported findings appear to support the idea that low frequency oscillations are associated with behavioral approach and alpha with behavioral inhibition tendencies but these associations are only valid for induced responses to stimuli. PMID- 18396344 TI - In vitro kinetic analysis of carbohydrate and aromatic amino acid metabolism of different members of the human colon. AB - The carbohydrate and aromatic amino acid metabolism of several species related to the human colon was investigated into more detail. Therefore, in vitro fermentations were performed, with different carbohydrate sources, during which several aromatic amino acids were added to the fermentation medium. Shifts in end product formation in response to the available nutrients were observed for all strains tested. The major part of amino acid degradation occurred after depletion of the carbohydrates. Moreover, it was shown that Bifidobacterium strains are capable of degrading aromatic amino acids in the absence of carbohydrates. The excretion of certain intermediates of the aromatic amino acid metabolism was observed for a strain of Clostridium clostridioforme, after which they were metabolized again during a later stage of fermentation. This implies that cross feeding on degradation products of aromatic amino acids, albeit within the same species, can occur in the human colon. PMID- 18396346 TI - Chromatin changes induced by lamin A/C deficiency and the histone deacetylase inhibitor trichostatin A. AB - Recent studies have shown that histone code dictates the type and structure of chromatin. Bearing in mind the importance of A-type lamins for chromatin arrangement, we studied the effect of trichostatin A (TSA)-induced histone hyperacetylation in lamin A/C-deficient (LMNA-/-) fibroblasts. Lamin A/C deficiency caused condensation of chromosome territories and the nuclear reorganization of centromeric heterochromatin, which was accompanied by the appearance of a chain-like morphology of HP1beta foci. Conversely, histone deacetylase (HDAC) inhibition induced de-condensation of chromosome territories, which compensated the effect of lamin A/C deficiency on chromosome regions. The amount of heterochromatin in the area associated with the nuclear membrane was significantly reduced in LMNA-/- cells when compared with lamin A/C-positive (LMNA+/+) fibroblasts. TSA also decreased the amount of peripheral heterochromatin, similarly as lamin A/C deficiency. In both LMNA+/+ and LMNA-/- cells, physically larger chromosomes were positioned more peripherally as compared with the smaller ones, even after TSA treatment. Our observations indicate that lamin A/C deficiency causes not only reorganization of chromatin and some chromatin-associated domains, but also has an impact on the extent of chromosome condensation. As HDAC inhibition can compensate the lamin A/C dependent chromatin changes, the interaction between lamins and specifically modified histones may play an important role in higher-order chromatin organization, which influences transcriptional activity. PMID- 18396345 TI - The human granulocyte nucleus: Unusual nuclear envelope and heterochromatin composition. AB - The human blood granulocyte (neutrophil) is adapted to find and destroy infectious agents. The nucleus of the human neutrophil has a segmented appearance, consisting of a linear or branched array of three or four lobes. Adequate levels of lamin B receptor (LBR) are necessary for differentiation of the lobulated nucleus. The levels of other components of the nuclear envelope may also be important for nuclear shape determination. In the present study, immunostaining and immunoblotting procedures explored the levels of various components of the nuclear envelope and heterochromatin, comparing freshly isolated human neutrophils with granulocytic forms of HL-60 cells, a tissue culture model system. In comparison to granulocytic HL-60 cells, blood neutrophil nuclear envelopes contain low-to-negligible amounts of LBR, lamins A/C, B1 and B2, LAP2beta and emerin. Surprisingly, a "mitotic" chromosome marker, H3(S10)phos, is elevated in neutrophil nuclei, compared to granulocytic HL-60 cells. Furthermore, neutrophil nuclei appear to be more fragile to methanol fixation, than observed with granulocytic HL-60 cells. Thus, the human neutrophil nucleus appears to be highly specialized, possessing a paucity of nuclear envelope-stabilizing proteins. In consequence, the neutrophil nucleus appears to be very malleable, supporting rapid migration through tight tissue spaces. PMID- 18396348 TI - Pustulosis acuta generalisata with joint involvement in an HLA-A2- and HLA-B35 positive patient. AB - Pustulosis acuta generalisata (PAG) is a rare poststreptococcal disease of the skin, which has been reported in children and adults after streptococcal throat infection. Herein, we report on the case of a 47-year-old woman with typical clinical and histologic findings of PAG emerging after a pharyngeal infection in whom inflammatory joint-involvement developed. The patient was found to be HLA-A2 and HLA-B35 positive. Whereas HLA-B35 might be associated with pustular skin diseases, HLA-A2 is a risk factor for the development of rheumatoid arthritis. PMID- 18396347 TI - IgG anti-laminin-332 autoantibodies are present in a subset of patients with mucous membrane, but not bullous, pemphigoid. AB - BACKGROUND: Antiepiligrin cicatricial pemphigoid is a mucosal-predominant subepidermal blistering disease associated with an increased relative risk of cancer. In contrast to prior reports showing that anti-laminin (L)-332 autoantibodies are a reliable marker for patients with antiepiligrin cicatricial pemphigoid, a recent report suggested that as many as 40% of patients with bullous pemphigoid (BP) have IgG reactive with this laminin isoform. OBJECTIVE: We sought to determine whether patients with BP possess circulating IgG anti-L 332 autoantibodies. METHODS: Sera from 100 adults with BP were analyzed by indirect immunofluorescence testing of intact skin, immunoblot studies of human keratinocyte (HK) extracts, and a new L-332 enzyme-linked immunosorbent assay. Sera showing reactivity suggestive of anti-L-332 autoantibodies in these assays were further analyzed in immunoblot studies of HK extracellular matrix and immunoprecipitation studies of biosynthetically radiolabeled HK extracts. RESULTS: IgG from all patients with BP bound intact epidermal basement membrane by indirect immunofluorescence microscopy and immunoblotted bullous pemphigoid antigen-1, -2, or both in HK extracts. None of these sera immunoblotted L-332 in HK extracts, although 13 did score above the cut point of a new IgG(4) L-332 enzyme-linked immunosorbent assay (sensitivity = 0.91, specificity = 0.98, Youden index = 0.89). Further analysis of sera from these 13 patients found: (1) all had IgG that bound the epidermal side of 1 mol/L NaCl split skin by indirect immunofluorescence microscopy; (2) none immunoblotted L-332 purified from HK extracellular matrix; and (3) none immunoprecipitated L-332 from biosynthetically radiolabeled HK extracts. LIMITATIONS: The basis of false-positive enzyme-linked immunosorbent assay determinations for anti-L-332 IgG among patients with BP is unknown. CONCLUSION: Anti-L-332 autoantibodies remain a reliable marker for patients with antiepiligrin cicatricial pemphigoid. PMID- 18396349 TI - Use of multilocus sequence typing for the investigation of colonisation by Candida albicans in intensive care unit patients. AB - A prospective study was performed to determine the prevalence of candidal colonisation on the general intensive care unit at a large teaching hospital. Colonisation with Candida spp. was found to be common, occurring in 79% of patients on the unit. C. albicans was the commonest species, colonising 64% of patients, followed by C. glabrata (18%) and C. parapsilosis (14%). Most of the members of staff tested carried Candida spp. at some point, although carriage appeared to be transient. C. parapsilosis was the most commonly isolated species from staff hands, whereas C. albicans was the most commonly isolated species from the mouth. The molecular epidemiology of C. albicans was investigated using Ca3 typing and multilocus sequence typing (MLST). MLST proved to be a reproducible typing method and a useful tool for the investigation of the molecular epidemiology of C. albicans. The results of the molecular typing provided evidence for the presence of an endemic strain on the unit, which was isolated repeatedly from patients and staff. This finding suggests horizontal transmission of C. albicans on the unit though it may also reflect the relative frequency of C. albicans strain types colonising patients on admission. This study has important implications for the epidemiology of systemic candidal infections. PMID- 18396350 TI - Ghrelin, des-acyl ghrelin and obestatin: three pieces of the same puzzle. AB - The major active product of ghrelin gene is a 28-amino acid peptide acylated at the serine 3 position with an octanoyl group, called simply ghrelin. Ghrelin has a multiplicity of physiological functions, affecting GH release, food intake, energy and glucose homeostasis, gastrointestinal, cardiovascular, pulmonary and immune function, cell proliferation and differentiation and bone physiology. Nevertheless, recent developments have shown that ghrelin gene can generate various bioactive molecules besides ghrelin, mainly des-acyl ghrelin and obestatin, obtained from alternative splicing or from extensive post translational modification. Although their receptors have not yet been identified, they have already proven to be active, having intriguingly subtle but opposite physiological actions to ghrelin. This suggests the existence of a novel endocrine system with multiple effector elements which not only may have opposite actions but may regulate the action of each other. In this review, we summarize the steps which lead to the production of the different ghrelin gene products and examine the most significant differences between them in terms of structure and actions. PMID- 18396351 TI - Electrophysiological properties and gap junction coupling of striatal astrocytes. AB - The striatum is the biggest nucleus of the basal ganglia and receives input from almost all cortical regions, substantia nigra and the thalamus. Striatal neuronal circuitry is well characterized, but less is known about glial physiology. To this end, we evaluated astrocyte electrophysiological properties using whole-cell patch-clamp recording in dorsal striatal brain slices from P15 to P21 rat. The majority of cells (95%) were passive astrocytes that do not express any detectable voltage-gated channels. Passive astrocytes were subcategorized into three groups based on time-dependent current properties. The observed proportion of the different astrocyte subtypes did not change within the age range evaluated here, but was modulated during reduction of specific conductances and gap junction coupling. Striatal astrocytes were extensively interconnected and closure of gap junctions with octanol (1mM), carbenoxolone (100 microM) or increased intracellular calcium (2mM), significantly altered intrinsic properties. When simultaneously blocking potassium channels and gap junction coupling almost no passive conductance was detected, implying that the major currents in striatal astrocytes derive from potassium and gap junction conductance. Uncoupling of the syncytium reduced currents activated in response to a hyperpolarizing pulse, suggesting that changes in gap junction coupling alters astrocyte electrophysiological responses. Our findings indicate that the prevalent gap junction coupling is vital for astrocyte function in the striatum, and that whole-cell recordings will be distorted by currents activated in neighboring cells. PMID- 18396352 TI - Synthesis of some novel N4-(naphtha[1,2-d]thiazol-2-yl)semicarbazides as potential anticonvulsants. AB - A series of N(4)-(naphtha[1,2-d]thiazol-2-yl)semicarbazides were designed and synthesized to meet the structural requirements essential for anticonvulsant activity. Anticonvulsant activity was determined after intraperitoneal (i.p.) administration to mice by maximal electroshock (MES) and subcutaneous pentylenetetrazole (scPTZ)-induced seizure tests and minimal motor impairment was determined by rotorod test. A majority of the compounds exhibited significant anticonvulsant activity after intraperitoneal administration. Some of the selected compounds were evaluated orally in rats for activity in scPTZ test at several time points (50 mg/kg). The most active compounds carry bromo, fluoro and nitro substituents at 4-position in the phenyl ring. The biochemical estimations of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) from brain homogenate not only clearly implicated the role of free radicals in PTZ-induced convulsion but also explained the possible mechanism of protective effect of semicarbazides, through the reduced formation of MDA and increased formation of SOD and GSH-Px. PMID- 18396353 TI - Synthesis and evaluation of novel bis-pyridinium oximes as reactivators of DFP inhibited acetylcholinesterase. AB - A series of novel bis-pyridinium oximes connected by bis-methoxymethyl benzene, 1,4-bis-methoxymethyl (cis)-but-2-ene and 1,4-bis-methoxymethyl but-2-yne linkers were synthesized and their in vitro reactivation efficacy was evaluated against diisopropyl phosphorofluoridate (DFP) inhibited acetylcholinesterase (AChE) and compared with the established antidote 2-PAM and obidoxime. However, the best reactivation was observed with the standard oxime 2-PAM. The reactivation efficacy of 1,3-dimethoxymethyl benzene bis-[4,4'-(hydroxyiminomethyl) pyridinium] dichloride (3d) and 1,4-dimethoxy but-2-ene bis-[4,4' (hydroxyiminomethyl) pyridinium] dichloride (3g) was comparable with that of obidoxime, another standard antidote. PMID- 18396354 TI - Design, synthesis, and evaluation of benzophenone derivatives as novel acetylcholinesterase inhibitors. AB - Starting from a structure-based drug design, new acetylcholinesterase inhibitors were designed and synthesized as analogues of donepezil. The compounds were composed by an aromatic function and a tertiary amino moiety connected by a suitable spacer. In particular, the benzophenone nucleus and the N,N benzylmethylamine function were selected. The easily accessible three-step synthesis of these compounds resulted to be significantly less difficult and expensive than that of donepezil. Several compounds possess anti-cholinesterase activity in the order of micro and sub-micromolar. Particularly, compounds 1 and 10 were the most potent inhibitors of the series. PMID- 18396355 TI - In-vitro regeneration of sarin inhibited electric eel acetylcholinesterase by bis pyridinium oximes bearing xylene linker. AB - A series of bis-pyridinium oximes connected by xylene linker were synthesized and their in-vitro reactivation potential was evaluated against acetylcholinesterase (AChE) inhibited by nerve agent, sarin. Among the synthesized compounds, alpha,alpha'xylene-bis-[3,3'-(hydroxyiminomethyl) pyridinium] dibromide (3b) was found to be most potent reactivator for AChE inhibited by sarin. The oxime 3b exhibits 34% regeneration of inhibited AChE, in comparison to 20 and 15% regeneration by 2-PAM and obidoxime, respectively, at a concentration of 10(-4) M within 10 min. PMID- 18396356 TI - Design and synthesis of calpain inhibitory 6-pyridone 2-carboxamide derivatives. AB - Excessive calpain activation contributes to serious cellular damage in many pathological conditions. The involvement of mu-calpain in neurological disorders such as, stroke and Alzheimer's disease has attracted considerable interest in the use of calpain inhibitors as therapeutic agents. 6-Pyridone 2-carboxamides derived from ketoamides were synthesized as conformationally constrained structures resembling the well known peptidic mu-calpain inhibitor, MDL 28,170, and their mu-calpain inhibitory activities were evaluated. Of the compounds synthesized, compound 2a, which has a primary amide at warhead region of the inhibitor most potently inhibited mu-calpain with an IC(50) value of 2.81+/-1.26 microM, which is ca. 40-fold less than that of MDL 28,170. PMID- 18396357 TI - Simple synthesis of carbon-11 labeled styryl dyes as new potential PET RNA specific, living cell imaging probes. AB - A new type of styryl dyes have been developed as RNA-specific, live cell imaging probes for fluorescent microscopy technology to study nuclear structure and function. This study was designed to develop carbon-11 labeled styryl dyes as new probes for biomedical imaging technique positron emission tomography (PET) imaging of RNA in living cells. Precursors (E)-2-(2-(1-(triisopropylsilyl)-1H indol-3-yl)vinyl)quinoline (2), (E)-2-(2,4,6-trimethoxystyryl)quinoline (3) and (E)-4-(2-(6-methoxyquinolin-2-yl)vinyl)-N,N-diemthylaniline (4), and standards styryl dyes E36 (6), E144 (7) and F22 (9) were synthesized in multiple steps with moderate to high chemical yields. Precursor 2 was labeled by [(11)C]CH(3)OTf, trapped on a cation-exchange CM Sep-Pak cartridge following a quick deprotecting reaction by addition of (n-Bu)(4)NF in THF, and isolated by solid-phase extraction (SPE) purification to provide target tracer [(11)C]E36 ([(11)C]6) in 40-50% radiochemical yields, decay corrected to end of bombardment (EOB), based on [(11)C]CO(2). The target tracers [(11)C]E144 ([(11)C]7) and [(11)C]F22 ([(11)C]9) were prepared by N-[(11)C]methylation of the precursors 3 and 4, respectively, using [(11)C]CH(3)OTf and isolated by SPE method in 50-70% radiochemical yields at EOB. The specific activity of the target tracers [(11)C]6, [(11)C]7 and [(11)C]9 was in a range of 74-111GBq/mumol at the end of synthesis (EOS). PMID- 18396358 TI - Synthesis and potential anticonvulsant activity of new N-3-substituted 5,5 cyclopropanespirohydantoins. AB - Thirteen new 5-cyclopropanespirohydantoins with various N-3 substituents were synthesized and their pharmacological activity was determined with the objective to better understand their structure-activity relationship (SAR) for anticonvulsant activity. The anticonvulsant effects of these compounds were evaluated by maximal electroshock seizure (MES) test and subcutaneous pentylenetetrazole (scPTZ) test models in mice. All compounds substituted with cyclopropyl group at fifth position of hydantoin ring showed better protection against MES test. Compounds 5b, 5d, 5e, 5g and 5j were found to be the most potent compounds of this series and compared with the reference drug phenytoin sodium in MES test. Compound 5j also showed equipotent activity with the standard drug sodium valproate at the doses of 20 and 40 mg kg(-1) in scPTZ test. PMID- 18396359 TI - Synthesis and QSAR modeling of 2-acetyl-2-ethoxycarbonyl-1-[4(4'-arylazo)-phenyl] N,N-dimethylaminophenyl aziridines as potential antibacterial agents. AB - The present communication deals with the synthesis of a series of 2-acetyl-2 ethoxycarbonyl-1-[4(4'-arylazo)-phenyl]-N,N-dimethylaminophenyl aziridines. The compounds were synthesized in excellent yields (70-80%) and the structures were established on the basis of consistent IR, (1)H NMR and elemental analysis data. The purity has been ascertained by chromatographic resolution using acetic acid- toluene (6:4v/v) as binary eluent. All the compounds have been tested for their antimicrobial activity against a representative panel of bacteria i.e. Bacillus subtilis, Escherichia coli, Pseudomonas diminuta and Staphylococcus aureus using azinomycin as reference drug. All the synthesized compounds were found to exhibit profound antimicrobial activity. PMID- 18396360 TI - Long-term immunogenicity of a single dose of acellular pertussis vaccine in paediatric health-care workers. AB - OBJECTIVE: Monitor the long-term immunogenicity of a single dose of acellular pertussis vaccine in health-care workers. DESIGN: German health-care workers and child-care workers received a single dose of a monovalent acellular pertussis vaccine (PAC-Merieux) in an open-label study. Blood samples were taken before (n=261), 4 weeks after (n=246), 1 year (n=187), 2 years (n=53), 3 years (n=134) and 4 years (n=37) after vaccination. IgG- and IgA-anti-pertussis-toxin (PT), IgG and IgA-anti-filamentous hemagglutinin (FHA), and IgG-anti-pertactin (PRN) were measured by ELISA. RESULTS: Of all subjects, 97.1%, 99.2% and 97.2% had an antibody response to PT, FHA and PRN, respectively. Four weeks after vaccination the median titres of IgG antibodies to PT, FHA and PRN were 314, 785 and 84 EU/l, respectively, and all vaccinees had an immune response to at least one pertussis antigen. IgA-anti-PT and IgA-anti-FHA responses were found in 63.4% and 96.3% of subjects with a median titre of 30 and 196 EU/ml, respectively. The titre of IgG anti-PT decreased slowly with a median concentration of 76, 71, 71 and 63 EU/ml after 1, 2, 3 and 4 years, respectively. Secondary titre increases were observed in 0.5%, 3.3%, 5.6% and 12.5% of the vaccinees 1, 2, 3, and 4 years after vaccination. CONCLUSION: In German health paediatric care workers long-lasting immune responses with high antibody levels could be induced by a single dose of acellular pertussis vaccine. A renewed contact with B. pertussis antigens resulted in a measurable immune response to PT between 0.5% (1 year p.v.) and 12.5% (4 years p.v.). PMID- 18396361 TI - Randomized, placebo-controlled, double-blind trial of the Na-ASP-2 hookworm vaccine in unexposed adults. AB - Necator americanus Ancylostoma Secreted Protein-2 (Na-ASP-2) is a leading larval stage hookworm vaccine candidate. Recombinant Na-ASP-2 was expressed in Pichia pastoris and formulated with Alhydrogel. In a phase 1 trial, 36 healthy adults without history of hookworm infection were enrolled into 1 of 3 dose cohorts (n=12 per cohort) and randomized to receive intramuscular injections of either Na ASP-2 or saline placebo. Nine participants in the first, second and third cohorts were assigned to receive 10, 50 and 100 microg of Na-ASP-2, respectively, on study days 0, 56 and 112, while 3 participants in each cohort received placebo. The most frequent adverse events were mild-to-moderate injection site reactions; in 8 participants these were delayed and occurred up to 10 days after immunization. No serious adverse events occurred. Anti-Na-ASP-2 IgG endpoint titers as determined by ELISA increased from baseline in all vaccine groups and peaked 14 days after the third injection, with geometric mean titers of 1:7066, 1:7611 and 1:11,593 for the 10, 50 and 100 microg doses, respectively, compared to <1:100 for saline controls (p<0.001). Antibody titers remained significantly elevated in all vaccine groups until the end of the study, approximately 8 months after the third vaccination. In vitro stimulation of PBMCs collected from participants with Na-ASP-2 resulted in robust proliferative responses in those who received vaccine, which increased with successive immunizations and remained high in the 50 and 100 microg dose groups through the end of the study. This first trial of a human hookworm vaccine demonstrates that the Na-ASP-2 vaccine is well-tolerated and induces a prolonged immune response in adults not exposed to hookworm, justifying further testing of this vaccine in an endemic area. PMID- 18396362 TI - Surface modified liposomes for nasal delivery of DNA vaccine. AB - The aim of the present work was to investigate the potential utility of glycol chitosan coated liposomes as nasal vaccine delivery vehicle for eliciting viral specific humoral mucosal and cellular immune responses. Plasmid pRc/CMV-HBs(S) encapsulated liposomes were prepared by dehydration-rehydration method and subsequently coated with glycol chitosan by simple incubation method. Liposomes were then characterized for their size, surface charge, entrapment efficiency, and ability to protect encapsulated DNA against nuclease digestion and for their mucoadhesiveness. The liposomes were then administered to mice in order to study their feasibility as nasal vaccine carriers. The developed liposomes possessed +9.8 mV zeta potential and an average vesicle size less than 1 microm and entrapment efficiency of approximately 53%. Following intranasal administration, glycol chitosan coated liposomes elicited humoral mucosal and cellular immune responses that were significant as compared to naked DNA justifying the potential advantage of mucosal vaccination in the production of local antibodies at the sites where pathogens enters the body. PMID- 18396363 TI - Tracking historical lead pollution in the coastal area adjacent to the Yangtze River Estuary using lead isotopic compositions. AB - The rapid economic development in the Yangtze River Delta (YRD), China in the last three decades has had a significant impact on the environment of the East China Sea (ECS). Lead isotopic compositions of a (210)Pb dated sediment core collected from the coastal ECS adjacent to the Yangtze River Estuary were analyzed to track the Pb pollution in the region. The baseline Pb concentration in the coastal ECS sediments before the industrialization in China was 32 microg g(-1), and the corresponding 206Pb/207Pb ratio was 1.195. The high-resolution profiles of Pb flux and 206Pb/207Pb ratios had close relationships with the economic development and the history of the use of leaded gasoline in China, and they were clearly different from those of most European countries and United States. PMID- 18396364 TI - Low toxicant concentrations decrease the frequency of fast-growing seedlings at high densities of annual baby's breath (Gypsophila elegans). AB - Very low toxicant concentrations reduce the frequency of most vital seedlings in dense stands in vitro. PMID- 18396365 TI - An evaluation of the risks of lung cancer and mesothelioma from exposure to amphibole cleavage fragments. AB - Amphiboles are hydrated mineral silicates five of which occur in asbestiform habits as asbestos grunerite (amosite) asbestos, riebeckite (crocidolite) asbestos, anthophyllite asbestos, tremolite asbestos and actinolite asbestos] and non-asbestiform habits (grunerite, riebeckite, anthophyllite, tremolite and actinolite). The asbestiform varieties are characterized by long, thin fibers while non-asbestiform varieties such as cleavage fragments form short fibers with larger widths. The U.S. regulatory method for counting asbestos fibers (aspect ratio > or = 3:1, length > or = 5 microm) does not distinguish between asbestos and cleavage fragments. The method biases toward increased counts of non asbestiform cleavage fragments compared to long, thin asbestos fibers. One consequence of this regulatory approach is that workers can be erroneously classified as exposed to concentrations of asbestos (asbestiform amphiboles) above the U.S. 0.1 f/mL exposure standard when in fact they are not exposed to asbestos at all but non-asbestiform amphibole cleavage fragments. Another consequence is that the known carcinogenic effects of asbestos may be falsely attributed to non-asbestiform amphibole cleavage fragments of the same mineral. The purpose of this review is to assess whether amphibole cleavage fragments pose the same risk of lung cancer and mesothelioma characteristic of amphibole asbestos fibers. We identified three groups of workers exposed to non-asbestiform amphiboles: two groups exposed to grunerite (Homestake gold miners and taconite miners) and one group exposed to industrial talc containing non-asbestiform tremolite and anthophyllite in St. Lawrence County, NY. In addition to assessing strength of association and exposure-response trends in the non-asbestiform amphibole cohorts, comparisons were also made with cohorts exposed to the asbestiform counterpart (positive control) and cohorts exposed to the mineral (e.g. talc) that does not contain amphiboles (negative controls). The cohorts exposed to non-asbestiform amphiboles had no excesses of lung cancer or mesothelioma. Similar results were observed in the negative control groups, in stark contrast to the excess risks of asbestos-related disease found in the asbestos cohorts. The only possible exception is the twofold increased risk of lung cancer where exposure was to industrial talc containing cleavage fragments of tremolite and anthophyllite. However, this risk is not considered attributable to the talc or amphibole cleavage fragments for several reasons. A similar increased risk of lung cancer was found in Vermont talc workers, studied in the same time period. Their exposure was to relatively pure talc. There was no relationship between lung cancer mortality and exposure measured as mg/m(3)years and years worked. A case-control study reported that all the lung cancer cases were smokers (or former smokers) and attributed the excess to smoking. There were two mesothelioma cases among the NY State talc workers exposed to cleavage fragments of tremolite and anthophyllite, but talc is not a plausible cause because of too short latency and potential for previous asbestos exposure. The positive controls of tremolite asbestos and anthophyllite asbestos exposed workers showed excess risks of both lung cancer and mesothelioma and positive exposure-response trends. St. Lawrence, NY talc does not produce mesotheliomas in animals while amphibole asbestos does. In sum, the weight of evidence fully supports a conclusion that non-asbestiform amphiboles do not increase the risk of lung cancer or mesothelioma. PMID- 18396366 TI - Pathways to depression: the impact of neighborhood violent crime on inner-city residents in Baltimore, Maryland, USA. AB - Crime and neighborhood disorder may negatively impact the health of urban residents. Neighborhoods with high levels of violent crime may also increase residents' risk of experiencing violence. Most studies supporting the assertion that neighborhood disorder impacts mental health have used residents' own ratings of their neighborhoods. The present study examines the relationships among block group level crime, perceived neighborhood disorder, violence experienced in the neighborhood, and depression. The sample comprising the current and former drug users (n=786) nested in 270 block groups within Baltimore, Maryland, USA. Using path analysis, we tested the hypothesis that neighborhood violent crime has a direct impact on experiences of violence. Also, we hypothesized that neighborhood violence had a direct and indirect impact on depressive symptoms. Results support a model in which violence is associated with psychological distress through perceptions of neighborhood disorder, and through experiences of violence. We conclude that community and structural level interventions are needed to decrease neighborhood crime and improve residents' perception of their neighborhood. PMID- 18396367 TI - The two facets of electronic care surveillance: an exploration of the views of older people who live with monitoring devices. AB - Scholars are increasingly questioning the notion that electronic surveillance merely constrains individuals' liberty and privacy. However, illustrations of alternative perspectives are few and there is a need for empirical research exploring the actual experience of surveilled subjects. This study, carried out in Sweden, seeks to offer a nuanced account of how senior citizens experience electronic care surveillance in relation to their privacy. It is based on in depth interviews with 17 seniors who have participated in a telemonitoring project and who have experience of being continuously activity monitored in their own homes. The findings suggest that senior citizens can perceive electronic care surveillance as freeing and as protecting their privacy, as it enables them to continue living in their own home rather than moving to a nursing home. One individual, however, experienced a privacy violation and the surveillance service was interrupted at her request. This illustrates the importance of built-in possibilities for subjects to exit such services. In general, the study highlights that e-surveillance can be not only constraining but also enabling. Hence, it supports the view of the dual nature of surveillance. The study also illustrates the agency of the surveilled subject, extending the argument that various agents actually participate in the construction of surveillance practices. It analyzes the indirect role and responsibility of the surveilled subject, and thereby questions the traditional roles ascribed to the agents and targets of surveillance. PMID- 18396369 TI - Prospective randomized, double-blinded, placebo-controlled trial of preoperative etoricoxib for pain relief in uterine fractional curettage under paracervical block. AB - OBJECTIVE: To evaluate the analgesic efficacy of preoperative etoricoxib combined with paracervical nerve block in patients who underwent uterine fractional curettage. STUDY DESIGN: This double-blinded, randomized, placebo-controlled trial included 80 women who underwent uterine fractional curettage under paracervical block. Forty women were randomly assigned to etoricoxib 120 mg and 40 women to the placebo. The main outcome measure was the intensity of pain measured using the visual analog pain score during and after the procedure. Student's t-tests, Mann-Whitney U-test, and Chi-squared test were used for statistical analysis. RESULTS: The intensity of pain during operative procedure in the etoricoxib group was lower than in the placebo group (median visual analog pain scores (interquartile range) 48 (43-64) vs. 61 (51-72), P=0.001) The amount of postoperative acetaminophen used in the etoricoxib group was also lower than in the placebo group (2.2+/-1.7 vs. 3.2+/-1.7 tablets, P=0.011). We found no significant adverse effects in this study. CONCLUSION: The preoperative administration of 120 mg oral etoricoxib can slightly reduce pain during fractional curettage under paracervical block. However, the degree of pain reduction by this treatment has no clinical importance. PMID- 18396370 TI - Precipitation assessment in wastewater treatment plants operated for biological nutrient removal: a case study in Murcia, Spain. AB - The Murcia Este Wastewater Treatment Plant is the largest wastewater treatment plant in Murcia (Spain). The plant operators have continuously found pipe blockage and accumulation of solids on equipment surfaces during the anaerobic digestion and post-digestion processes. This work studies the precipitation problems in the Murcia Este Wastewater Treatment Plant in order to locate the sources of precipitation and its causes from an exhaustive mass balance analysis. The DAF thickener and anaerobic digester mass balances suggest that most of the polyphosphate is released during excess sludge thickening. Despite the high concentrations achieved in the thickened sludge, precipitation does not occur at this point due to the low pH. The increases in ammonium and pH during anaerobic digestion cause precipitation to take place mainly inside the digesters and in downstream processes. This study shows that 50.7% of the available phosphate is fixed in the digester of which 52.0% precipitates as ammonium struvite, 39.2% precipitates as hydroxyapatite and the remaining 8.8% is adsorbed on the surface of the solids. Thermodynamic calculations confirm the precipitation of struvite and hydroxyapatite and also confirm that potassium struvite does not precipitate in the anaerobic digesters. PMID- 18396371 TI - Role of Fgf receptor 2c in adipocyte hypertrophy in mesenteric white adipose tissue. AB - Fgf receptor 2c (Fgfr2c) was expressed in mature adipocytes of mouse white adipose tissue (WAT). To examine the role of Fgfr2c in mature adipocytes, we generated adipocyte-specific Fgfr2 knockout (Fgfr2 CKO) mice. The hypertrophy impairment of adipocytes in the mesenteric WAT but not in the subcutaneous WAT and decreased plasma free fatty acid (FFA) levels were observed in Fgfr2 CKO mice. Although the expression of genes involved in adipocyte differentiation and lipid metabolism in the mesenteric WAT was essentially unchanged, the expression of uncoupling protein 2 potentially involved in energy dissipation was significantly increased. Among potential Fgf ligands for Fgfr2c, Fgf9 was preferentially expressed in the mesenteric WAT. The present findings indicate that Fgfr2c potentially activated by Fgf9 plays a role in the adipocyte hypertrophy in the mesenteric WAT and FFA metabolism and/or energy dissipation in the mesenteric WAT might be involved in the hypertrophy impairment. PMID- 18396372 TI - Could cytokines levels or adhesion molecules expression be predictor of IFN-beta treatment response in multiple sclerosis patients? AB - OBJECTIVE: Some patients with relapsing-remitting multiple sclerosis (RRMS) do not respond to treatment with interferon-beta and continue to have relapses and new enhancing lesions on MRI. The markers which would predict the treatment response are still not known. The objectives of the study were to compare cytokines levels (IFN-gamma, IL-4, IL-6, IL-10) and expression of adhesion molecules before and during treatment in responders and nonresponders to IFN-beta treatment. METHODS: Twenty-nine patients with RRMS were enrolled in the study. Cytokine levels were evaluated by ELISA in supernatants of IONO/PMA activated PBMC cultures (IFN-gamma, IL-4, IL-6, IL-10), and by flow cytometry (intracellular IFN-gamma, IL-4 and IL-2R expression) before and during treatment. Expression of adhesion molecules (VLA-4, ICAM-1) was evaluated by flow cytometry (CD49+ and CD54+) before and during treatment. RESULTS: Only 9 of 29 patients were responders to treatment according to definition (no relapse in the first 2 years of treatment). We found significant differences in the expression of IL-2R after 1 month of treatment, intracellular IFN-gamma after 6 months of treatment and IL-10 level in nonactivated PBMC cultures after 1 week of treatment. CONCLUSION: We concluded that the differences we had found between responders and nonresponders are most probably incidental and not predictive of treatment response. Our study shows that cytokines levels and expression of adhesion molecules cannot be used as markers for treatment response. PMID- 18396373 TI - Tertiary treatment of textile wastewater with combined media biological aerated filter (CMBAF) at different hydraulic loadings and dissolved oxygen concentrations. AB - An up-flow biological aerated filter packed with two layers media was employed for tertiary treatment of textile wastewater secondary effluent. Under steady state conditions, good performance of the reactor was achieved and the average COD, NH(4)(+)-N and total nitrogen (TN) in the effluent were 31, 2 and 8mg/L, respectively. For a fixed dissolved oxygen (DO) concentration, an increase of hydraulic loading resulted in a decrease in substrate removal. With the increase of hydraulic loadings from 0.13 to 0.78m(3)/(m(2)h), the removal efficiencies of COD, NH(4)(+)-N and TN all decreased, which dropped from 52 to 38%, from 90 to 68% and from 45 to 33%, respectively. In addition, the results also confirmed that the increase of COD and NH(4)(+)-N removal efficiencies resulted from the increase of DO concentrations, but this variation trend was not observed for TN removal. With the increase of DO concentrations from 2.4 to 6.1mg/L, the removal efficiencies of COD and NH(4)(+)-N were 39-53% and 64-88%, whenas TN removal efficiencies increased from 39 to 42% and then dropped to 35%. PMID- 18396375 TI - AFM picking-up manipulation of the metaphase chromosome fragment by using the tweezers-type probe. AB - We have studied the development of a new procedure based on atomic force microscopy (AFM) for the analysis of metaphase chromosome. The aim of this study was to obtain detailed information about the specific locations of genes on the metaphase chromosome. In this research, we performed the manipulation of the metaphase chromosome by using novel AFM probes to obtain chromosome fragments of a smaller size than the ones obtained using the conventional methods, such as glass microneedles. We could pick up the fragment of the metaphase chromosome dissected by the knife-edged probe by using our tweezers-type probe. PMID- 18396374 TI - Cytokine profile in human skin in response to experimental inflammation, noxious stimulation, and administration of a COX-inhibitor: a microdialysis study. AB - Animal studies have documented a critical role for cytokines in cell signaling events underlying inflammation and pain associated with tissue injury. While clinical reports indicate an important role of cytokines in inflammatory pain, methodological limitations have made systematic human studies difficult. This study examined the utility of a human in vivo bioassay combining microdialysis with multiplex immunoassay techniques for measuring cytokine arrays in tissue. The first experiment measured cytokines in interstitial fluid collected from non inflamed and experimentally inflamed skin (UVB). The effects of noxious heat on cytokine release were also assessed. The second experiment examined whether anti hyperalgesic effects of the COX-inhibitor ibuprofen were associated with decreased tissue levels of the pro-inflammatory cytokines IL-1 beta and IL-6. In the first experiment, inflammation significantly increased IL-1 beta, IL-6, IL-8, IL-10, G-CSF, and MIP-1 beta. Noxious heat but not experimental inflammation significantly increased IL-7 and IL-13. In the second experiment, an oral dose of 400 and 800 mg ibuprofen produced similar anti-hyperalgesic effects suggesting a ceiling effect. Tissue levels of IL-1 beta and IL-6 were not affected after the 400mg dose but decreased significantly (44+/-32% and 38+/-13%) after the 800 mg dose. These results support the utility of explored method for tracking cytokines in human tissue and suggest that anti-hyperalgesic and anti-inflammatory effects of ibuprofen are at least partially dissociated. The data further suggest that high clinical doses of ibuprofen exert anti-inflammatory effects by down regulating tissue cytokine levels. Explored human bioassay is a promising tool for studying the pathology and pharmacology of inflammatory and chronic pain conditions. PMID- 18396368 TI - Erythropoietin: elucidating new cellular targets that broaden therapeutic strategies. AB - Given that erythropoietin (EPO) is no longer believed to have exclusive biological activity in the hematopoietic system, EPO is now considered to have applicability in a variety of nervous system disorders that can overlap with vascular disease, metabolic impairments, and immune system function. As a result, EPO may offer efficacy for a broad number of disorders that involve Alzheimer's disease, cardiac insufficiency, stroke, trauma, and diabetic complications. During a number of clinical conditions, EPO is robust and can prevent metabolic compromise, neuronal and vascular degeneration, and inflammatory cell activation. Yet, use of EPO is not without its considerations especially in light of frequent concerns that may compromise clinical care. Recent work has elucidated a number of novel cellular pathways governed by EPO that can open new avenues to avert deleterious effects of this agent and offer previously unrecognized perspectives for therapeutic strategies. Obtaining greater insight into the role of EPO in the nervous system and elucidating its unique cellular pathways may provide greater cellular viability not only in the nervous system but also throughout the body. PMID- 18396376 TI - In vitro effect of herbal products against Histomonas meleagridis. AB - The ban on chemotherapeutics against Histomonas meleagridis in the European Union has left turkey producers without an effective treatment against histomonosis. It has encouraged the development of alternative control methods amongst which are a number of herbal products. In the present study the in vitro effect of four herbal products against H. meleagridis was examined. After suspension of the herbal products in Dwyer's culture medium used for subculturing of a viable culture, parasite growth was monitored by frequently counting the histomonads until 72h of incubation. The solid products Aromabiotictrade mark, Enteroguardtrade mark, and Protophyt SPtrade mark as well as the positive control products dimetridazole and Histostat-50trade mark, were tested in twofold serial dilutions ranging in concentration from 400 to 12.5mug/mL, while the liquid product Protophyt Btrade mark was tested in concentrations ranging from 0.24% to 0.008% (v/v). The herbal products Enteroguardtrade mark and Protophyt Btrade mark as well as dimetridazole and Histostat-50trade mark, two chemotherapeutics with known antihistomonal activity, inhibited the growth of H. meleagridis, while no antihistomonal effect was found for Aromabiotictrade mark and Protophyt SPtrade mark. PMID- 18396377 TI - Gender-specific behavioral and immunological alterations in an animal model of autism induced by prenatal exposure to valproic acid. AB - Autism is a severe behavioral disorder characterized by pervasive impairments in social interactions, deficits in verbal and non-verbal communication, and stereotyped behaviors, with a four times higher incidence in boys than in girls. The core symptoms are frequently accompanied by a spectrum of neurobehavioral and immunological derangements, including: aberrant sensitivity to sensory stimulation, anxiety, and decreased cellular immune capacity. Recently, a new potential rodent model of autism induced by prenatal exposure to valproic acid (VPA rats) has been proposed. In order to determine if gender has an influence on alterations observed in VPA rats, male and female rats have been evaluated in a battery of behavioral, immunological, and endocrinological tests. A plethora of aberrations has been found in male VPA rats: lower sensitivity to pain, increased repetitive/stereotypic-like activity, higher anxiety, decreased level of social interaction, increased basal level of corticosterone, decreased weight of the thymus, decreased splenocytes proliferative response to concanavaline A, lower IFN-gamma/IL-10 ratio, and increased production of NO by peritoneal macrophages. Female VPA rats exhibited only increased repetitive/stereotypic-like activity and decreased IFN-gamma/IL-10 ratio. Sexual dimorphism characteristics for measured parameters have been observed in both groups of animals, except social interaction in VPA rats. Our results confirm existence of similarities between the observed pattern of aberrations in VPA rats and features of disturbed behavior and immune function in autistic patients, and suggest that they are gender-specific, which is intriguing in light of disproportion in boys to girls ratio in autism. PMID- 18396378 TI - Birth characteristics and childhood malignant central nervous sytem tumors: the ESCALE study (French Society for Childhood Cancer). AB - BACKGROUND: Determining the role of pre- and perinatal factors in the aetiology of childhood malignant central nervous (CNS) tumors, using data from the French national case-control study, ESCALE. METHODS: ESCALE included all children in France less than 15 years old with a diagnosis of acute leukaemia, lymphoma, malignant CNS tumor, or neuroblastoma (2003-2004). In all, 209 malignant CNS tumor cases (80% of the eligible cases) and 1681 population-based controls (71%) were included using quotas ensuring frequency matching with the cases by age and gender. Case and control mothers were interviewed using a standardised telephone interview, which elicited birth characteristics, congenital malformation, maternal reproductive history, and use of assisted reproductive technologies for the index child. RESULTS: The cases and controls did not differ in terms of gestational age at birth, birth weight, birth order, breastfeeding, or parental age at birth. There was no association between assisted reproduction for the index child and malignant CNS tumor (OR=1.1 [0.6-2.2]). A positive association between a maternal history of one miscarriage and malignant CNS tumor was observed (OR=1.4 [1.0-2.0], p<0.05), especially for glial cell tumors (other glioma: OR=2.0 [1.1-3.6]). CONCLUSION: The results suggest a possible association between a maternal history of one miscarriage and the risk of malignant CNS tumor. PMID- 18396379 TI - Summation and subtraction using a modified autoshaping procedure in pigeons. AB - A modified autoshaping paradigm (significantly different from those previously reported in the summation literature) was employed to allow for the simultaneous assessment of stimulus summation and subtraction in pigeons. The response requirements and the probability of food delivery were adjusted such that towards the end of training 12 of 48 trials ended in food delivery, the same proportion as under testing. Stimuli (outlines of squares of three sizes and colors: A, B, and C) were used that could be presented separately or in any combination of two or three stimuli. Twelve of the pigeons (summation groups) were trained with either A, B, and C or with AB, BC, and CA, and tested with ABC. The remaining 12 pigeons (subtraction groups) received training with ABC but were tested with A, B, and C or with AB, BC, and CA. These groups were further subdivided according to whether stimulus elements were presented either in a concentric or dispersed manner. Summation did not occur; subtraction occurred in the two concentric groups. For interpretation of the results, configural theory, the Rescorla-Wagner model, and the composite-stimulus control model were considered. The results suggest different mechanisms responsible for summation and subtraction. PMID- 18396380 TI - Genomic structure of nitric oxide synthase in the terrestrial slug is highly conserved. AB - Nitric oxide (NO) is a key molecule in olfactory information processing across animal species. To gain insight into the genetic basis of NO generation, we investigated the genomic structure of nitric oxide synthase (NOS) in the terrestrial slug Limax because slugs use olfaction as their primary food detection system. The full length cDNA of limNOS encodes a protein consisting of 1632 amino acids that has a PSD-95/discs-large/ZO-1 (PDZ) domain in its N terminus and 6 other cofactor-binding domains. The limNOS gene consists of 33 exons and spans at least 107 kb of the genome. Almost all the exon-intron boundaries are conserved in Limax and human nNOS and the organization of the Limax genome is more similar to that of humans than to Drosophila, indicating that there was an accelerated evolution of the Drosophila genome during evolution. These results imply that there was a highly conservative selective pressure imposed on NOS gene structure during the evolution of mollusks and vertebrates. PMID- 18396381 TI - Development of hyperactivity after hearing loss in a computational model of the dorsal cochlear nucleus depends on neuron response type. AB - Cochlear damage can change the spontaneous firing rates of neurons in the dorsal cochlear nucleus (DCN). Increased spontaneous firing rates (hyperactivity) after acoustic trauma have been observed in the DCN of rodents such as hamsters, chinchillas and rats. This hyperactivity has been interpreted as a neural correlate of tinnitus. In cats, however, the spontaneous firing rates of DCN neurons were not significantly elevated after acoustic trauma. Species-specific spontaneous firing rates after cochlear damage might be attributable to differences in the response types of DCN neurons: In gerbils, type III response characteristics are predominant, whereas in cats type IV responses are more frequent. To address the question of how the development of hyperactivity after cochlear damage depends on the response type of DCN neurons, we use a computational model of the basic circuit of the DCN. By changing the strength of two types of inhibition, we can reproduce salient features of the responses of DCN neurons. Simulated cochlear damage, which decreases the activity of auditory nerve fibers, is assumed to activate homeostatic plasticity in projection neurons (PNs) of the DCN. We find that the resulting spontaneous firing rates depend on the response type of DCN PNs: PNs with type III and type IV-T response characteristics may become hyperactive, whereas type IV PNs do not develop increased spontaneous firing rates after acoustic trauma. This theoretical framework for the mechanisms and circumstances of the development of hyperactivity in central auditory neurons might also provide new insights into the development of tinnitus. PMID- 18396382 TI - [Nuclear receptor PPAR and hepatology: pathophysiological and therapeutical aspects]. AB - In last few years, the topic of nuclear receptor has been developed in the field of hepatology allowing envisaging therapeutic strategies for the most frequent chronic liver diseases. Peroxysome proliferator-activated receptors (PPAR) contribute to wide physiological processes within the liver such as lipid/glucid metabolisms, inflammatory response, cell differenciation and cell cycle. In vitro experiments and animal studies showed that PPARalpha discloses anti-inflammatory property and PPARgamma discloses anti-inflammatory, antifibrogenic and antiproliferative properties in the liver. Main available agonists are fibrates (PPARalpha) used for 20 years in cases of lipid metabolism abnormalities and glitazones (PPARgamma) used since 2000 for type 2 diabetes. In terms of therapy, animal studies and human trials have been conducted in steatopathies. However, clinicians have to be aware of potential specific side effects related to glitazones especially on cardiovascular system. PMID- 18396383 TI - Role of p38 MAPK and pro-inflammatory cytokines expression in glutamate-induced neuronal death of neonatal rats. AB - Pro-inflammatory cytokines TNF-alpha, IL-1beta and IL-6 rises significantly during neuronal damage and activate the signaling p38 MAPK pathway, which is involved in the apoptotic (AP) neuronal death. Systemic administration of glutamate as monosodium salt (MSG) to newborn animals induces neuronal death, however whether neurons die by AP or necrosis through MAPK p38 pathway activation it is unknown. In this study, TNF-alpha, IL-1beta and IL-6 expression levels, AP neuronal death and cellular type that produces TNF-alpha was also identified in the cerebral cortex (CC) and striatum (St) of rats at 8, 10, and 14 days of age after neonatal exposure to MSG. TNF-alpha production and AP neuronal death was significantly increased in the CC at PD8-10, and in the St in all ages studied by excitotoxicity effect induced with MSG. This effect was completely inhibited by SB203580 (p38 inhibitor) in both regions studied. TNF-alpha, IL-1beta and IL-6 RNAm increased after MSG administration, whereas SB203580 did not modify their expression. These data indicates that neuronal death induced by excitotoxicity appears to be mediated through p38 signaling pathway activated by TNF-alpha and their inhibition may have an important neuroprotective role as part of anti inflammatory therapeutic strategy. PMID- 18396384 TI - Salvage surgery after failed chemoradiotherapy in squamous cell carcinoma of the esophagus. AB - AIMS: To investigate the survival benefit and preoperative risk factors for hospital mortality of salvage surgery in esophageal cancer patients who had locoregional residual/recurrent tumor after definitive chemoradiotherapy. METHODS: We retrospectively reviewed the esophageal cancer patients who presented at our hospital from 1997 to 2004. Forty-seven patients who had squamous cell cancer and developed locoregional recurrent/persistent disease after primary definitive chemoradiotherapy were elected. Twenty-seven of them received salvage esophagectomy (group 1) and the other 20 underwent non-operative treatment only (group 2). In order to assess the surgery-related mobility and mortality in group 1, 191 patients who received neoadjuvant chemoradiotherapy followed by operation during the same time period were also enrolled (group 3). RESULTS: The 5-year overall survival of group 1 patients was 25.4%. In contrast, all of the patients in the group 2 died within 16.7 months. The difference was statistically significant (p=0.0029). In comparison with group 3, group 1 patients had significantly more surgery-related complications and hospital mortality. In univariate analysis for preoperative risk factors, a low albumin or hemoglobulin level was associated with high hospital mortality in group 1 (p=0.004 and 0.003, respectively). After multivariate analysis, only the low albumin level remained borderline significance. As for disease specific survival after salvage surgery, R0 resection was the only independent prognosticator (p=0.049). CONCLUSION: Salvage surgery provides survival benefit in esophageal cancer patients with locoregional persistent or recurrent disease after primary definitive chemoradiotherapy. Preoperative albumin and hemoglobulin levels are associated with hospital mortality and may aid in selecting suitable patient for salvage surgery. PMID- 18396385 TI - Retinol up-regulates the receptor for advanced glycation endproducts (RAGE) by increasing intracellular reactive species. AB - Retinol (vitamin A) and other retinoids have been suggested to exert an important antioxidant function in biological systems, besides their more established role as regulators of cell growth and differentiation. On the other hand, many authors have recently observed pro-oxidant activities of vitamin A and other retinoids in vitro and in vivo, resulting in cell death and/or transformation associated to increased oxidative damage. However, the mechanisms by which retinol causes oxidative stress are still not fully understood. Receptors for advanced glycation endproducts (RAGE) have been recently implied as promoters and/or amplifiers of oxidant-mediated cell death induced by diverse agents, and increased RAGE expression is observed in conditions related to unbalanced production of reactive species, such as in atherosclerosis and neurodegeneration. In the present work, we observed that retinol supplementation increases RAGE protein expression in cultured Sertoli cells, and antioxidant co-treatment reversed this effect. Retinol-increased RAGE expression was observed only at concentrations that induce intracellular reactive species production, as assessed by the DCFH assay. These results indicate that retinol is able to increase RAGE expression by an oxidant dependent mechanism, and suggest that RAGE signaling may be involved in some of the deleterious effects observed in some retinol-supplementation therapies. PMID- 18396386 TI - Are metabolic indices different between drug-naive first-episode psychosis patients and healthy controls? AB - OBJECTIVE: To compare glucose and lipid metabolism parameters between drug-naive first-episode psychosis (FEP) patients with a diagnosis of schizophrenia spectrum disorder and healthy controls matched for age, ethnicity, and gender. METHOD: Baseline evaluations of fasting glucose and lipid metabolism parameters and the oral glucose tolerance test were performed with FEP patients (n=38), having no more than 10 days of cumulative exposure to antipsychotic medication, and normal community controls (n=36). Analysis of variance (ANOVA) was conducted to examine between group differences. RESULTS: FEP patients did not show a higher prevalence of the precursors to diabetes (impaired fasting glucose, impaired glucose tolerance, insulin resistance), and no significant difference in beta-cell function or lipid profile measures, compared to healthy controls. FEP patients showed a higher waist to hip ratio compared to controls. CONCLUSIONS: FEP patients having a schizophrenia spectrum disorder do not differ from healthy controls, in their baseline measures of glucose and lipid metabolites, nor in the prevalence of diabetes or its precursors, before (or close to) the onset of antipsychotic treatment. PMID- 18396387 TI - Specific linguistic and pragmatic deficits in Italian patients with schizophrenia. AB - OBJECTIVE: Verbal communication impairments are prominent features of schizophrenia. The grammatical and pragmatic components of expressive and receptive verbal abilities were systematically examined, for the first time, in Italian patients with schizophrenia. Indeed, most of the language literature is composed of studies on English speaking people. METHOD: Elicited narrative production, and syntactic and pragmatic receptive abilities were analyzed in a cohort of 37 patients with schizophrenia and 37 healthy controls. Furthermore, a conversational speech production task was administered to an age- and gender matched subset of this population. The level of significance was set at p or =3 per year. Patients with other respiratory diseases, renal failure, and congestive heart failure were excluded. Serum magnesium was assayed at the time of admission by the colorimetric method. Logistic regression analysis was used to find independent risk factors for readmission. RESULTS: The mean age of patients was 71.9 (+/-10.9 standard deviation (SD)) years. Fifty-seven were females. Ninety percent were current or ex-smokers. The median duration of time to next admission was 108 days (range 2-842). A total of 87 patients were readmitted at least once during the first year of follow-up, while 5% died; 23% had frequent readmissions; 85% had received pneumococcal vaccine within 5 years; and 29% received influenza vaccine in the current season. Frequency of readmissions was not influenced by the administration of inhaled or oral steroids, and diuretics at the time of discharge. Vaccination did not protect against frequent readmissions. The sole predictor of frequent readmissions was serum magnesium level (1.77+/-0.19 vs. 1.86+/-0.24mEq/L; adjusted odds ratio 0.003, 95% Confidence intervals <0.001-0.55; p=0.03). CONCLUSIONS: Lower serum magnesium level independently predicts readmission for AECOPD. This is an easily modifiable risk factor. PMID- 18396394 TI - Effect of compost application rate on carbon degradation and retention in soils. AB - We investigated the effect of a single compost application at two rates (50 and 85Mgha(-1)) on carbon (C) degradation and retention in an agricultural soil cropped with maize after 150d. We used both C mass balance and soil respiration data to trace the fate of compost C. Our results indicated that compost C accumulated in the soil after 150d was 4.24Mgha(-1) and 6.82Mg C ha(-1) for 50 and 85Mg ha(-1) compost rate, respectively. Compost C was sequestered at the rate of 623 and 617g C kg(-1) compost TOC for 50 and 85Mgha(-1) compost dose, respectively. These results point to a linear response between dose of application and both C degradation and retention. The amount of C sequestered was similar to the total recalcitrant C content of compost, which was 586g C kg(-1) compost TOC, indicating that, probably, during the short experiment, the labile C pool of compost (414g C kg(-1) of compost TOC) was completely degraded. Soil respiration measured at different times during the crop growth cycle was stable for soils amended with compost (CO2 flux of 0.96+/-0.11g CO2 m(-2) h(-1) and 1.07+/-0.10g CO2 m(-2) h(-1), respectively, for 50 and 85Mgha(-1)), whereas it increased in the control. The CO2 flux due to compost degradation only, though not statistically significant, was always greatest for the highest compost doses applied (0.22+/-0.40g CO2 m(-2) h(-1) and 0.33+/-0.25g CO2 m(-2) h(-1) for the 50 and 85Mgha(-1) compost dose, respectively). This seems to confirm the highest C degradation for the 85Mgha(-1) compost dose as a consequence of the presence of more labile C. Unlike other studies, the results show a slight increase in the fraction of carbon retained with the increase in compost application rate. This could be due to the highly stable state of the compost prior to application, although it could also be due to sampling uncertainty. Further investigations are needed to better explain how the compost application rate affects carbon sequestration, and how characterization into labile and recalcitrant C can predict the amount of C sequestered in the soil. PMID- 18396395 TI - Landfills in Jiangsu province, China, and potential threats for public health: leachate appraisal and spatial analysis using geographic information system and remote sensing. AB - Waste disposal is of growing environmental and public health concern in China where landfilling is the predominant method of disposal. The assessment of potential health hazards posed by existing landfills requires sound information, and processing of a significant amount of spatial data. Geographical information system (GIS) and remote sensing (RS) are valuable tools for assessing health impacts due to landfills. The aims of this study were: (i) to analyze the leachate and gas emissions from landfills used for domestic waste disposal in a metropolitan area of Jiangsu province, China, (ii) to investigate remotely-sensed environmental features in close proximity to landfills, and (iii) to evaluate the compliance of their location and leachate quality with the relevant national regulations. We randomly selected five landfills in the metropolitan areas of Wuxi and Suzhou city, Jiangsu province, established a GIS database and examined whether data were in compliance with national environmental and public health regulations. The leachates of the sampled landfills contained heavy metals (Pb, As, Cr(6+) and Hg) and organic compounds in concentrations considered harmful to human health. Measured methane concentrations on landfill surfaces were low. Spatial analysis of the location of landfills with regard to distance from major water bodies, sensible infrastructure and environmental conditions according to current national legislation resulted in the rejection of four of the five sites as inappropriate for landfills. Our results call for rigorous evaluation of the spatial location of landfills in China that must take into consideration environmental and public health criteria. PMID- 18396396 TI - The clinical significance of lymphangiogenesis and angiogenesis in non-small cell lung cancer patients. AB - BACKGROUND: Angiogenesis and lymphangiogenesis have been reported to affect malignant phenotype. METHOD: We investigated 147 patients with non-small cell lung cancer (NSCLC). Immunohistochemistry using D2-40 was performed to evaluate lymphatic vessel density (LVD), including Micro-LVD (without lumen), Tubal-LVD (with lumen) and lymphatic vessel invasion (LVI). The intratumoural microvessel density (MVD) was evaluated by CD-34 immunostaining. The expressions of vascular endothelial growth factor-A (VEGF-A) and VEGF-C were also studied. RESULTS: Lymphangiogenesis was significantly associated with Micro-LVD (p=0.0003). The VEGF-C expression was significantly associated with the Micro-LVD (p=0.0057). In contrast, the VEGF-A expression was significantly associated with the MVD (p=0.0092). The survival was significantly lower in patients with Micro-LVD-high tumours than in patients with Micro-LVD-low tumours (p=0.0397). Survival was also significantly lower in patients with MVD-high tumours than in patients with MVD low tumours (p=0.0334). A multivariate analysis demonstrated that the Micro-LVD (p=0.0363) and the MVD (p=0.0232) were independent prognostic factors for NSCLC patients. CONCLUSIONS: Lymphangiogenesis, specifically Micro-LVD and angiogenesis are independently associated with a poor prognosis in NSCLC patients. PMID- 18396397 TI - Terminology: the historical perspective, evolution and current usage--room for confusion? AB - Modern palliative care started with St Christopher's Hospice in 1967 and was initially regarded as 'terminal care'. This served as a template for a developing model of multidisciplinary clinical care, teaching and research. A decade later, several hospital Palliative Care Teams were established and different terms were used to describe them. An evidence base developed slowly and a medical subspeciality was established, known as Palliative Medicine. Over the last two decades we have seen an expansion in non-hospice palliative care. The terms used to describe this care have been variable and inconsistent. Our challenges in progress involve establishing clear terminology and an evolving improved evidence base, along with a realisation that there are large gaps in patient care. PMID- 18396398 TI - Chemical compositions and larvicidal activities of leaf essential oils from two eucalyptus species. AB - In the current study, the mosquito larvicidal activity of leaf essential oils and their constituents from two eucalyptus species (Eucalyptus camaldulensis and Eucalyptus urophylla) against two mosquito species, Aedes aegypti and Aedes albopictus, was investigated. In addition, the chemical compositions of the leaf essential oils were analyzed using gas chromatography-mass spectrometry. Results from the larvicidal tests revealed that essential oil from the leaves of E. camaldulensis had an excellent inhibitory effect against both A. aegypti and A. albopictus larvae. The 12 pure constituents extracted from the two eucalyptus leaf essential oils were also tested individually against two mosquito larvae. Among the six effective constituents, alpha-terpinene exhibits the best larvicidal effect against both A. aegypti and A. albopictus larvae. Results of this study show that the leaf essential oil of E. camaldulensis and its effective constituents might be considered as a potent source for the production of fine natural larvicides. PMID- 18396399 TI - The design and synthesis of diaryl ether second generation HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs) with enhanced potency versus key clinical mutations. AB - Using a combination of traditional Medicinal Chemistry/SAR analysis, crystallography, and molecular modeling, we have designed and synthesized a series of novel, highly potent NNRTIs that possess broad antiviral activity against a number of key clinical mutations. PMID- 18396400 TI - Synthesis and DNA binding ability of cyclam-amino acid conjugates. AB - N-Substituted cyclam-amino acid conjugates have been synthesised both in solution and on the solid phase. The DNA binding affinity of these species has been studied: the nature of the amino acid strongly influences the change in melting temperature suggesting that simple cyclam-peptide conjugates could interact with DNA in a highly selective manner. PMID- 18396401 TI - New targeting system for antimycotic drugs: beta-glucosidase sensitive amphotericin B-star poly(ethylene glycol) conjugate. AB - A new targeting potentially intravenous conjugate Amphotericin B (AMB)-star poly(ethylene glycol) (sPEG) (M=25,160) has been synthesized and characterized. It contains a beta-d-glucopyranoside molecular switch which is sensitive to beta glucosidases (E.C.3.2.1.21). The beta-glucosidase-catalyzed release of AMB from the polymeric carrier was proved in vitro by means of spectrophotometry and HPLC. PMID- 18396402 TI - 4th UK spinal muscular atrophy (SMA) researchers network meeting. PMID- 18396403 TI - Trigeminal schwannomas: experience with 68 cases. AB - Trigeminal schwannomas (TS) are rare. Only a couple of series involving a large number of cases have been reported. In the present study we aimed to analyse the clinical characteristics of TS, the surgical approaches used to treat TS, and the outcomes for patients undergoing surgical treatment for TS via retrospective analysis of departmental records. Data for 68 patients treated for TS in the Department of Neurosurgery at the All India Institute of Medical Sciences between January 1993 and December 2005 were analysed. Most patients were in the fourth decade of life, with the duration of symptoms ranging from 1 month to 13 years. Twenty-nine TSs were classified as type A, 13 as type B and 26 as type C, depending upon size. A skull base approach was used in every surgically treated case. Of the 46 patients for whom radiological follow-up data were available, complete tumour excision was achieved in 35 cases (76%). Follow-up ranged from 3 months to 12 years (mean 62 months). One patient died and nine (15%) had permanent morbidity in the form of corneal opacity (5) or facial (2) or trochlear (2) nerve palsy. We conclude that trigeminal neuromas are best treated by total surgical resection, which yields acceptable results with low rates of mortality and permanent morbidity. PMID- 18396404 TI - Synthesis and cytotoxic activity of gamma-aryl substituted alpha-alkylidene-gamma lactones and alpha-alkylidene-gamma-lactams. AB - A series of 5-aryl-3-alkylidenedihydrofuran-2(3H)-ones 6a-g'' and 11a,b as well as 5-aryl-3-methylidenepyrrolidin-2-ones 10a-c and 12 were synthesized starting from 4-aryl-2-diethoxyphosphoryl-4-oxobutanoates 3a-g. Reaction sequence includes reduction or reductive amination of the carbonyl group, lactonization or lactamization step and finally the Horner-Wadsworth-Emmons olefination of aldehydes using thus obtained 5-aryl-3-diethoxyphosphoryl-3,4-dihydrofuran-2(5H) ones 5a-g'' or 5-aryl-3-diethoxyphosphorylpyrrolidin-2-ones 9a-c. Furanones 6 and 11, as well as pyrrolidinones 10 and 12, were evaluated in vitro against mouse leukemia cell line L-1210 and two human leukemia cell lines HL-60 and NALM-6. Several of the obtained furanones proved to be very potent against all three cell lines with IC(50) values lower than 6 microM. Structure-activity relationships of these compounds, as well as 5-alkyl or 5-arylmethyl-3-methylidenedihydrofuran 2(3H)-ones 13a-e, previously obtained in our laboratory, are discussed. PMID- 18396405 TI - Podophyllotoxin directly binds a hinge domain in E2 of HPV and inhibits an E2/E7 interaction in vitro. AB - Podophyllotoxin (PT), a strong cytotoxic agent from berberidaceae, has been known to inhibit tubulin polymerization. Although PT has been used for developing anticancer drugs as one of seed compounds, clinical treatment by itself has been unsuccessful because of the side effects, except one example in the treatments of warts. In this study, we screened peptides binding to PT with T7 phage display clonings in order to obtain more information about molecular mechanism of the action. A selected phage clone has a specific amino acid sequence to be SVPSRRRPDGRTHRSSRG. A homology search by protein database BLAST showed that this sequence had a similarity to a hinge domain (HD) of E2 protein in human papillomavirus (HPV) type 1a which is known to cause plantar warts. Surface plasmon resonance (SPR) analysis showed that PT bound to a recombinant HPV 1a E2 protein giving a K(D)=24.1microM which has compared with those of other domains of E2 protein. Also we demonstrated whether PT inhibited HD interaction or not. E7 protein of HPV has been known to be an oncoprotein and was reported to interact with HD of E2 protein. We demonstrated that an E2/E7 interaction was inhibited by the addition of PT in this report. And we showed the bindings of PT to other types of HPV. Our results suggest that PT is potential as a tool for clarifying the molecular mechanism of HPV. PMID- 18396406 TI - An improved synthesis of arsenic-biotin conjugates. AB - An amide linked conjugate of p-aminophenylarsine oxide and biotin is conveniently prepared in a one-pot procedure by the reaction of biotinyl chloride, formed in situ, with p-aminophenyldichloroarsine. The reaction of the arsine oxide-biotin conjugate with 1,2-ethanedithiol produces the stabilized dithiarsolane. These reagents are now readily available for a variety of applications. PMID- 18396407 TI - Gamma spectroscopic assay of soil samples from waste dump sites in Port Harcourt, Nigeria. AB - A lead shielded 7.6cmx7.6cm NaI (Tl) detector and a Canberra Multichannel Analyzer were employed for a gamma spectrometric assay of 100 soil samples from refuse dump sites in Port Harcourt city and environs. The massic activities of (40)K, (226)Ra and (228)Ra were determined and found to correspond to natural levels of some 10Bq/kg. No gamma-emitting anthropogenic radionuclides were detected. The mean radium equivalent is 44.2Bq/kg giving a dose rate of 24.6microSv/year. PMID- 18396409 TI - Efficient bacterial expression of fusion proteins and their selective processing by a recombinant Kex-1 protease. AB - A secreted, soluble variant of the Kex-1 endopeptidase from Kluyveromyces lactis has been produced and studied as a novel cleavage enzyme exhibiting high specificity for the Lys-Arg peptide. This highly selective, efficient enzyme is particularly adapted for use in manufacturing when a recombinant therapeutic protein, possessing its native N-terminus, has to be released in vitro from a bacterially-expressed fusion protein. In this paper, we describe the preparation of a Kex-1 variant using Saccharomyces cerevisiae and its application in the production of important therapeutic recombinant proteins such as human growth hormone, granulocyte colony-stimulating factor and interferon-alpha-2b. PMID- 18396410 TI - Assessment of the validity of the Biering-Sorensen test for measuring back muscle fatigue based on EMG median frequency characteristics of back and hip muscles. AB - The aims of the present study were (1) to investigate the differences in median frequency characteristics between back and hip muscles of healthy subjects during a Biering-Sorensen test, (2) to determine if the Biering-Sorensen test is a valid test for measuring back muscle fatigue, and (3) to standardise the Biering Sorensen test by using objective movement analysis when defining endurance time, and compare this to the original method based on tactile feedback. Twenty healthy subjects participated in this experiment. The electromyographic activity of eight back and hip muscles was bilaterally measured. In addition three-dimensional data of the lumbar region were collected with an ultrasound movement analysis system. Median frequencies were computed from the EMG power spectra. Two methods of determining the endurance time of the Biering-Sorensen test yielded highly correlated but significantly different normalized median frequency slope values (NMF(slope)). Significant differences in NMF(slope) values between several back and hip muscles could be demonstrated. Low to moderate correlation coefficients were shown between NMF(slope) values and endurance time. Multiple stepwise linear regression analyses revealed that only NMF(slope) of the thoracic part of the iliocostalis lumborum muscle could significantly predict the test endurance time. The findings of the present study support the validity of the Biering-Sorensen test for measuring back muscle fatigue. PMID- 18396411 TI - Correlations between short-time Fourier- and continuous wavelet transforms in the analysis of localized back and hip muscle fatigue during isometric contractions. AB - The aims of the current study were to examine the stationarities of surface electromyographic (EMG) signals obtained from eight bilateral back and hip muscles during a modified Biering-Sorensen test, and to investigate whether short time Fourier (STFT) and continuous wavelet transforms (CWT) provided similar information with regard to EMG spectral parameters in the analysis of localized muscle fatigue. Twenty healthy subjects participated in the study after giving their informed consent. Reverse arrangement tests showed that 91.6% of the EMG signal epochs demonstrated no significant trends (all p>0.05), meaning 91.6% of the EMG signal epochs could be considered as stationary signals. Pearson correlation coefficients showed that STFT and CWT in general provide similar information with respect to the EMG spectral variables during isometric back extensions, and as a consequence STFT can still be used. PMID- 18396408 TI - Cathepsin D--many functions of one aspartic protease. AB - For years, it has been held that cathepsin D (CD) is involved in rather non specific protein degradation in a strongly acidic milieu of lysosomes. Studies with CD knock-out mice revealed that CD is not necessary for embryonal development, but it is indispensable for postnatal tissue homeostasis. Mutation that abolishes CD enzymatic activity causes neuronal ceroid lipofuscinosis (NCL) characterized by severe neurodegeneration, developmental regression, visual loss and epilepsy in both animals and humans. In the last decade, however, an increasing number of studies demonstrated that enzymatic function of CD is not restricted solely to acidic milieu of lysosomes with important consequences in regulation of apoptosis. In addition to CD enzymatic activity, it has been shown that apoptosis is also regulated by catalytically inactive mutants of CD which suggests that CD interacts with other important molecules and influences cell signaling. Moreover, procathepsin D (pCD), secreted from cancer cells, acts as a mitogen on both cancer and stromal cells and stimulates their pro-invasive and pro-metastatic properties. Numerous studies found that pCD/CD level represents an independent prognostic factor in a variety of cancers and is therefore considered to be a potential target of anti-cancer therapy. Studies dealing with functions of cathepsin D are complicated by the fact that there are several simultaneous forms of CD in a cell-pCD, intermediate enzymatically active CD and mature heavy and light chain CD. It became evident that these forms may differently regulate the above-mentioned processes. In this article, we review the possible functions of CD and its various forms in cells and organisms during physiological and pathological conditions. PMID- 18396412 TI - Test-retest reliability of wavelet - and Fourier based EMG (instantaneous) median frequencies in the evaluation of back and hip muscle fatigue during isometric back extensions. AB - The present study aimed at assessing the test-retest reliability of wavelet - and Fourier derived (instantaneous) median frequencies of surface electromyographic (EMG) measurements of back and hip muscles during isometric back extensions. Twenty healthy subjects (10 males and 10 females) performed a modified Biering Sorensen test on two separate days, with a 1-week interval between the two tests. Surface EMG measurements were bilaterally performed from the latissimus dorsi, the thoracic and lumbar parts of the longissimus thoracis, the thoracic and lumbar parts of the iliocostalis lumborum, the multifidus, the gluteus maximus and the biceps femoris. In addition, three-dimensional kinematic data were recorded of the subjects' lumbar vertebrae. The (instantaneous) median frequencies were calculated from the EMG signals using continuous wavelet (IMDF) and short-time Fourier transforms (MDF). Linear regressions performed on the IMDF and MDF data as a function of time yielded slopes (IMDF(slope) and MDF(slope)) and intercepts (IMDF(init) and MDF(init)) of the regression lines. Test-retest reliability was assessed on the normalized slopes and intercept parameters by means of intraclass correlation coefficients (ICC) and standard errors of measurements expressed as percentages of the mean values (% SEM). The results of IMDF(slope) and MDF(slope) parameters indicated ICCs for back and hip muscles between .443 and .727 for IMDF(slope), values between .273 and .734 for MDF(slope), % SEM between 7.6% and 58.9% for IMDF(slope) and % SEM between 8.2% and 25.3% for MDF(slope), respectively. The ICCs for IMDF(init) and MDF(init) parameters varied between .376 and .907 for IMDF(init) and between .383 and .883 for MDF(init), and % SEM ranged from 2.7% to 6.3% for IMDF(init) and from 2.6% to 4.7% for MDF(init), respectively. These results indicate that both wavelet - and Fourier based (instantaneous) median frequency parameters generally are reliable in the analysis of back and hip muscle fatigue during a modified Biering-Sorensen test. PMID- 18396413 TI - Stimulus-response characteristics of motor evoked potentials and silent periods in proximal and distal upper-extremity muscles. AB - OBJECTIVE: To compare stimulus-response characteristics of both motor evoked potentials (MEP) and silent periods (SP) induced by transcranial magnetic stimulation (TMS) in proximal and distal upper-extremity muscles. METHODS: Stimulus-response curves of MEPs and SPs were obtained from the biceps brachii (BB) and abductor digiti minimi (ADM) muscles in 15 healthy participants. A nonlinear mixed model was used to fit the stimulus-response curves to a sigmoid Boltzmann function. RESULTS: Small residuals of the function were found for MEPs and SPs in both muscles. Higher maximal MEP amplitudes were found for the BB compared to the ADM (p<0.01). The active motor threshold to obtain a SP was less for the ADM compared to the BB (p<0.01). The slope parameter of the function of the SP duration was steeper and more variable in the ADM than in the BB (p<0.01). For the MEP amplitude no difference in active motor threshold and slope of the function was found between both muscles. CONCLUSIONS: Excitatory (MEP) and inhibitory (SP) effects of TMS differ between proximal arm and distal hand muscles in healthy participants. The adequate fit of our model suggests that this model can be used to study between and within subject changes in future studies. PMID- 18396414 TI - Amygdala responses to emotional faces in twins discordant or concordant for the risk for anxiety and depression. AB - BACKGROUND: Functional brain imaging studies have shown deviant amygdala responses to emotional stimuli in subjects suffering from anxiety and depressive disorder, but both hyperactivity and hypoactivity compared to healthy controls have been reported. To account for these discrepant findings, we hypothesize that genetic and environmental risk factors differently impact on amygdala functioning. METHODS: To test this hypothesis, we assessed amygdala responses to an emotional faces paradigm during functional magnetic resonance imaging in monozygotic twin pairs discordant for the risk of anxiety and depression (n=10 pairs) and in monozygotic twin pairs concordant for high (n=7 pairs) or low (n=15 pairs) risk for anxiety and depression. RESULTS: Main effects (all faces vs. baseline) revealed robust bilateral amygdala activity across groups. In discordant twins, increased amygdala responses were found for negatively valenced stimuli (angry/anxious faces) in high-risk twins compared to their low-risk co twins. In contrast, concordant high-risk pairs revealed blunted amygdala reactivity to both positive and negative faces compared with concordant low-risk pairs. Post-hoc analyses showed that these findings were independent of 5-HTTLPR genotype. CONCLUSIONS: Our findings indicate amygdala hyperactivity in subjects who are at high risk for anxiety and depression through environmental factors and amygdala hypoactivity in those at risk mainly through genetic factors. We suggest that this result reflects a difference in baseline amygdala activation in these groups. Future imaging studies on anxiety and depression should aim to avoid admixture of subjects who are at genetic risk with those at risk due to environmental factors. PMID- 18396415 TI - The BOLD response and vascular reactivity during visual stimulation in the presence of hypoxic hypoxia. AB - A disproportionate increase in cerebral blood flow (CBF) relative to the cerebral metabolic rate of oxygen (CMRO(2)), in response to neuronal activation, results in a decreased oxygen extraction fraction (OEF) and hence local 'hyperoxygenation'. The mismatch is the key 'physiological substrate' for blood oxygenation level dependent (BOLD) fMRI. The mismatch may reflect inefficient O(2) diffusion in the brain tissue, a factor requiring maintenance of a steep [O(2)] gradient between capillary bed and neural cell mitochondria. The aim of this study was to assess vascular responsiveness to reduced blood oxygen saturation, using both BOLD fMRI and the CBV-weighted vascular space occupancy (VASO)-dependent fMRI technique, during visual activation in hypoxic hypoxia. Our fMRI results show decreased amplitude and absence of initial sharp overshoot in the BOLD response, while VASO signal was not influenced by decreasing oxygen saturation down to 0.85. The results suggest that the OEF during visual activation may be different in hypoxia relative to normoxia, due to a more efficient oxygen extraction under compromised oxygen availability. The data also indicate that vascular reactivity to brain activation is not affected by mild hypoxia. PMID- 18396416 TI - Phylogeny of the Heelwalkers (Insecta: Mantophasmatodea) based on mtDNA sequences, with evidence for additional taxa in South Africa. AB - We examined the phylogeny of Mantophasmatodea from southern Africa (South Africa, Namibia) using approx. 1300 bp of mitochondrial DNA sequence data from the genes encoding COI and 16S. The taxon sample comprised multiple specimens from eight described species (Namaquaphasma ookiepense, Austrophasma rawsonvillense, A. caledonense, A. gansbaaiense, Lobatophasma redelinghuysense, Hemilobophasma montaguense, Karoophasma botterkloofense, K. biedouwense) and four undescribed species of Austrophasmatidae; three specimens of Sclerophasma paresisense (Mantophasmatidae); and two specimens of Praedatophasma maraisi and one of Tyrannophasma gladiator (not yet convincingly assigned to any family). For outgroup comparison a broad selection from hemimetabolous insect orders was included. Equally weighted parsimony analyses of the combined COI+16S data sets with gaps in 16S scored as a fifth character state supported Austrophasmatidae and all species and genera of Mantophasmatodea as being monophyletic. Most species were highly supported with 98-100% bootstrap/7-39 Bremer support (BS), but K. biedouwense had moderate support (87/4) and A. caledonense low support (70/1). Mantophasmatodea, Austrophasmatidae, and a clade Tyrannophasma gladiator+Praedatophasma maraisi were all strongly supported (99-100/12-25), while relationships among the two latter clades and Mantophasmatidae remain ambiguous. Concerning the relationships among genera of Austrophasmatidae, support values are moderately high for some nodes, but not significant for others. We additionally calculated the partitioned BS values of COI and 16S for all nodes in the strict consensus of the combined tree. COI and 16S are highly congruent at the species level as well as at the base of Mantophasmatodea, but congruence is poor for most intergeneric relationships. In forthcoming studies, deeper relationships in the order should be additionally explored by nuclear genes, such as 18S and 28S, for a reduced sample of specimens. PMID- 18396417 TI - Biomechanical testing of a new knotless suture anchor compared with established anchors for rotator cuff repair. AB - Various suture anchors are available for rotator cuff repair. For arthroscopic application, a knotless anchor was developed to simplify the intra-operative handling. We compared the new knotless anchor (BIOKNOTLESStrade mark RC; DePuy Mitek, Raynham, MA) with established absorbable and titanium suture anchors (UltraSorbtrade mark and Super Revo 5mmtrade mark; ConMed Linvatec, Utica, NY). Each anchor was tested on 6 human cadaveric shoulders. The anchors were inserted into the greater tuberosity. An incremental cyclic loading was performed. Ultimate failure loads, anchor displacement, and mode of failure were recorded. The anchor displacement of the BIOKNOTLESStrade mark RC (15.3 +/- 5.3 mm) after the first cycle with 75 N was significantly higher than with the two other anchors (Super Revo 2.1 +/- 1.6 mm, UltraSorb: 2.7 +/- 1.1 mm). There was no significant difference in the ultimate failure loads of the 3 anchors. Although the Bioknotlesstrade mark RC indicated comparable maximal pullout strength, it bares the risk of losing contact between the tendon-bone-interface due to a significantly higher system displacement. Therefore, gap formation between the bone and the soft tissue fixation jeopardizes the repair. Bioknotlesstrade mark RC should be used in the lateral row only when a double row technique for rotator cuff repair is performed, and is not appropriate for rotator cuff repair if used on its own. PMID- 18396418 TI - Osteochondritis dissecans involving the trochlea of the humerus treated with transplantation of tissue-engineered cartilage: a case report. PMID- 18396419 TI - Influence of sleep disturbance on quality of life of patients with epilepsy. AB - The frequency of sleep disturbances in patients with epilepsy and their impact on quality of life (QoL) have been documented in a few reports, and the results are conflicting. We identified 124 consecutive epilepsy out-patients who visited the epilepsy out-patient clinics at the University Hospital of Alexandroupolis, the AHEPA Hospital in Thessaloniki and the Aeginitio Hospital in Athens. We measured excessive daytime sleepiness (EDS) with the Epworth Sleepiness Scale (ESS), obstructive sleep apnea (OSA) with the Sleep Apnea scale of the Sleep Disorders Questionnaire (SA-SDQ), and insomnia with the Athens Insomnia Scale (AIS). We evaluated quality of life by the Quality of Life in Epilepsy Inventory (QOLIE 31). EDS was found in 16.9% (21/124) of epileptic patients, OSA in 28.2% (35/124), and insomnia in 24.6% (30/122). In multivariate analysis, we found that insomnia was an independent negative factor for Total score (p<0.001), Overall QoL (p=0.002), Emotional well-being (p<0.001), Energy/fatigue (p<0.001), Cognitive functioning (p=0.04) and Social functioning (p=0.03), and OSA only for Cognitive functioning (p=0.01). According to our findings, EDS, OSA, and insomnia are frequent in epileptic patients. Epileptic patients with sleep disturbance, mainly insomnia, have significant QoL impairment. PMID- 18396420 TI - The lateralizing value of IQ in mesiotemporal epilepsy: differences between patients with unitemporal and bitemporal epileptiform discharges. AB - PURPOSE: We investigated the lateralizing ability of intelligence scores in mesial temporal lobe epilepsy (MTLE) patients according to the distribution of interictal epileptiform discharges (IEDs). METHODS: This study enrolled 82 MTLE patients. All patients had preoperative neuropsychological evaluations, including Korean Wechsler Adult Intelligence Scale. Patients were categorized as having uni or bitemporal IEDs based on IEDs distribution (cutoff point, 90%). RESULTS: In patients with unitemporal IEDs, performance IQ (PIQ) was significantly lower in the right than in the left subgroup (89.6 vs. 99.4, p<0.05). Verbal IQ (VIQ)-PIQ discrepancy scores differed significantly between the left and right subgroups, being negative in the left and positive in the right subgroup. Based on multivariate analyses, two variables, right MTLE (p=0.042) and the unitemporal distribution of IEDs (p=0.030), were independently related to the VIQ-PIQ discrepancy of more than 10 points. About 47.4% of those with unitemporal IEDs had VIQ-PIQ discrepancies of greater than 10 points and the rate for correct lateralization was 77.8%. In patients with bitemporal IEDs, however, none of the intelligence scores showed evidence of correct lateralization. In patients with bitemporal IEDs, Full-scale IQ and PIQ were significantly lower in the left subgroup, and there was a significant difference in VIQ-PIQ discrepancy scores with the wrong direction. CONCLUSIONS: We found that intelligence scores had some lateralizing ability, but only in MTLE patients with unitemporal IEDs. PMID- 18396421 TI - Dual proline labeling protocol for individual "baseline" and "response" biosynthesis measurements in human articular cartilage. AB - OBJECTIVE: The heterogeneity of biosynthesis in human-derived cartilage explants poses a challenge to its use in experiments. The aim of this study was to determine the consistency with which two consecutive measures of biosynthesis could be made in individual human articular cartilage explants using a dual proline radiolabeling protocol. METHODS: Full-thickness cartilage explants were harvested from young bovine or human (total knee replacement) tibial plateaus. Two consecutive measurements of biosynthesis were obtained by measuring (3)H proline and (14)C-proline incorporation. Each sample's ratio of (14)C-/(3)H proline incorporation was computed. For comparison to traditional experimental designs, the (14)C-proline incorporation ratio was computed for adjacent cartilage samples. The number of samples needed to observe a change in the proline incorporation ratio of 10, 20, and 50% was determined for both methods. RESULTS: The dual-label ratio was consistent across samples from the same plateau [95% confidence interval (CI): +/-20% (human) and +/-30% (bovine) of median]. Adjacent human sample pairs had much greater variability in their (14)C-proline incorporation (95% CI: +/-50% of median). Adjacent bovine sample pairs had CIs that were similar in magnitude to those for the dual-label approach. In the human plateaus, ratio changes of 10, 20 and 50% could be detected using dramatically fewer samples than the adjacent pair method. For bovine samples, the two methods required a similar number of samples per group. CONCLUSION: The consistency of the dual-label approach may overcome the difficulties in studying the effects of interventions on biosynthesis in human cartilage in vitro. PMID- 18396422 TI - Hyperthermia in combination with oxidative stress induces autophagic cell death in HT-29 colon cancer cells. AB - The purpose of this study was to evaluate the mechanism of ROS-induced hyperthermic cell death in a colon cancer cell line. HT-29 colon cancer cells were exposed to heat (43 degrees C) in the presence of tert-butyl hydroperoxide (t-BOOH). t-BOOH combined with hyperthermia significantly decreased cell viability as compared with t-BOOH or hyperthermia alone. This decrease in cell numbers was associated with retardation in the S phase transit and not through apoptosis. Cell death was noted to be accompanied by specific features characteristic of autophagy: the presence of cytoplasmic autophagic vacuoles; autophagosome membrane association of microtubule-associated protein light chain 3; accumulation of acidic vesicular organelles; and increased incorporation of MDC in the autophagosome. Thermal sensitization through modulation of cellular ROS may represent a novel approach to increase the efficacy of hyperthermia as an anticancer modality. PMID- 18396423 TI - Characterization of mesenchymal stem cells isolated from the human umbilical cord. AB - Numerous papers have reported that mesenchymal stem cells (MSCs) can be isolated from various sources such as bone marrow, adipose tissue and others. Nonetheless it is an open question whether MSCs isolated from different sources represent a single cell lineage or if cells residing in different organs are separate members of a family of MSCs. Subendothelial tissue of the umbilical cord vein has been shown to be a promising source of MSCs. The aim of this study was to isolate and characterize cells derived from the subendothelial layer of umbilical cord veins as regards their clonogenicity and differentiation potential. The results from these experiments show that cells isolated from the umbilical cord vein displayed fibroblast-like morphology and grew into colonies. Immunophenotyping by flow cytometry revealed that the isolated cells were negative for the hematopoietic line markers HLA-DR and CD34 but were positive for CD29, CD90 and CD73. The isolated cells were also positive for survivin, Bcl-2, vimentin and endoglin, as confirmed by RT-PCR and immunofluorescence. These cells can be induced to differentiate into osteogenic and adipogenic cells, but a new finding is that these cells can be induced to differentiate into endothelial cells expressing CD31, vWF and KDR-2, and also form vessel-like structures in Matrigel. The differentiated cells stopped expressing survivin, thus showing a diminished proliferative potential. It can be assumed that the subendothelial layer of the umbilical cord vein contains a population of cells with the overall characteristics of MSCs, with the additional capability to transform into endothelial cells. PMID- 18396424 TI - Flow cytometric cell cycle analysis of cultured brown bear fibroblast cells. AB - The aim of this study was to assess by flow cytometry the cell cycle of brown bear fibroblast cells cultured under different growth conditions. Skin biopsies were taken in Cantabria (Spain) from a live, anaesthetized brown bear. DNA analysis was performed by flow cytometry following cell DNA staining with propidium iodide. Serum starvation increased (P<0.01) the percentage of G0/G1 phase cells (92.7+/-0.86) as compared to cycling cells (39.7+/-0.86) or cells cultured to confluency (87.3+/-0.86). DMSO included for 48h in the culture significantly increased (P<0.01) the percentage of G0/G1 phase of the cell cycle at all concentrations used and decreased percentages of S phase in a dose dependent fashion. Roscovitine increased the G0/G1 phase of the cell cycle (P<0.01) at 15microM concentration. Interestingly, the G2/M stage significantly increased at 30 and 50microM compared to the control and 15microM (P<0.02). The cell cycle of brown bear adult fibroblast cells can be successfully synchronized under a variety of culture conditions. PMID- 18396426 TI - The lytic effects of detergent sclerosants on erythrocytes, platelets, endothelial cells and microparticles are attenuated by albumin and other plasma components in vitro. AB - OBJECTIVE: To investigate the lytic effects of sodium tetradecyl sulphate (STS) and polidocanol (POL) on erythrocytes, platelets, endothelial cells and platelet derived microparticle (PDMP) formation in vitro and the potential protective effects of serum albumin and agents such as procaine. MATERIALS AND METHODS: The effects of sclerosants were studied in blood samples obtained from normal individuals. Absorbance densitometry was used to assess the lytic effects of sclerosants on blood cells and cultured human microvascular endothelial cells (HMEC) in plasma and in saline. PDMP were quantified by flow cytometry. RESULTS: Haemolysis occurred in whole blood at sclerosant concentrations greater than 0.25% for STS and above 0.45% for POL. Similar concentrations of both agents caused platelet and endothelial cell lysis. Both sclerosants released PDMP at low concentrations but destroyed PDMP at higher concentrations. Albumin significantly reduced the lytic effect of both sclerosants on all cells but had a greater inhibitory effect on POL. Protamine at 0.01% had a neutralising effect on STS, whereas procaine and lignocaine showed no such activity. CONCLUSIONS: Sclerosants at therapeutic concentrations lyse blood cells and endothelial cells in vitro. This effect is strongly reduced by serum albumin possibly contributing towards the low incidence of thromboembolic complications of sclerotherapy. PMID- 18396427 TI - Endovascular and open approaches to non-thrombosed popliteal aneurysm repair: a meta-analysis. AB - OBJECTIVE: Endovascular repair of popliteal artery aneurysms is a relatively new technique that is still undergoing evaluation. The aim of this study was to compare outcomes following open and endovascular approaches. METHODS: All published studies comparing outcomes following open and endovascular popliteal aneurysm were included. Endpoints included operative duration, length of stay, and postoperative complications including short-term patency rates. Outcomes were combined using a random-effects meta-analytical technique and differences assessed using odds ratios (OR), weighted mean difference (WMD) and log hazards ratio (HR). RESULTS: Three studies comprising 141 patients (37 endovascular; 104 open) were included. No significant differences in patient characteristics were seen. Operative duration was significantly longer for endovascular repair (WMD 120 minutes, p<0.001). Thirty day graft thrombosis (OR 5.05, p=0.06) and reintervention (OR 18.80, p=0.03) were more likely following endovascular repairs. Postoperative length of stay was shorter in the endovascular group (WMD- 3.9 days, p<0.001). There was no significant difference in long-term primary patency rates (HR 1.70, p=0.53). CONCLUSIONS: Endovascular repair of popliteal artery aneurysms offers similar medium-term benefits as an open repair. However, short-term graft thrombosis and reintervention rates are significantly greater. With the current technology it is difficult to justify endovascular treatment of popliteal aneurysms. PMID- 18396428 TI - Current perspectives on the diagnosis and epidemiology of Mycoplasma hyopneumoniae infection. AB - Mycoplasma hyopneumoniae is the principal aetiological agent of enzootic pneumonia (EP), a chronic respiratory disease that affects mainly finishing pigs. Although major efforts to control M. hyopneumoniae infection and its detrimental effects have been made, significant economic losses in pig production worldwide due to EP continue. M. hyopneumoniae is typically introduced into pig herds by the purchase of subclinically infected animals or, less frequently, through airborne transmission over short distances. Once in the herd, M. hyopneumoniae may be transmitted by direct contact from infected sows to their offspring or between pen mates. The 'gold standard' technique used to diagnose M. hyopneumoniae infection, bacteriological culture, is laborious and is seldom used routinely. Enzyme-linked immunosorbent assay and polymerase chain reaction detection methods, in addition to post-mortem inspection in the form of abattoir surveillance or field necropsy, are the techniques most frequently used to investigate the potential involvement of M. hyopneumoniae in porcine respiratory disease. Such techniques have been used to monitor the incidence of M. hyopneumoniae infection in herds both clinically and subclinically affected by EP, in vaccinated and non-vaccinated herds and under different production and management conditions. Differences in the clinical course of EP at farm level and in the efficacy of M. hyopneumoniae vaccination suggest that the transmission and virulence characteristics of different field isolates of M. hyopneumoniae may vary. This paper reviews the current state of knowledge of the epidemiology of M. hyopneumoniae infection including its transmission, infection and seroconversion dynamics and also compares the various epidemiological tools used to monitor EP. PMID- 18396429 TI - The pupillary light reflex and menace response in neonatal calves: the role of environmental isolation on development of the menace response. AB - The aim of this study was to investigate the development of the pupillary light reflex in neonatal calves and the effect on the development of the menace response of keeping them in individual stalls. Seventy-three Holstein calves were divided into test (n=44) and control (n=29) groups and the animals were kept in individual stalls for the first 32+/-4 and 73+/-5 days post partum, respectively. They were then moved to group pens. All calves in both groups were observed daily from the first day after birth until the pupillary light reflex and menace response had become well established. All 73 calves had a positive pupillary light reflex on the first day post partum. None of the animals developed a menace response during the time they were housed in individual stalls. Over 14 days (test group) and 12 days (control group) after the calves had been moved to the group pens they became positive to the menace response at ages 41+/-4 and 81+/-5 days, respectively. The Mann-Whitney test revealed a significant difference (P<0.001) in the ages of the calves with positive menace responses between groups. Group rearing may produce more visual threats to the eyes and the calves learn the menace response by closing the lids in response to sudden alarming movements. PMID- 18396430 TI - Atypical myopathy in grazing horses: a first exploratory data analysis. AB - Over the last decade, atypical myopathy (AM) in grazing horses has emerged in several European countries. An exploratory analysis was conducted to determine horse- and pasture-level indicators or factors associated with AM in Belgium. Belgian cases of AM confirmed by histology (n=57) were compared to their healthy co-grazing horses (n=77) and to pastured horses not involved with AM as controls (n=386). The pastures where confirmed cases were grazing (42 pastures; 38 sites; 44 incidences of AM) were compared with those of the controls (216 pastures; 96 sites; no incidence of AM). Statistically significant (P< or =0.05) exploratory variables, identified by means of adjusted odds ratios, suggested that indicators or factors associated with individual horses (young age, inactivity, body condition poor to normal), management practices (permanent pasturing, spreading of manure) and pasture characteristics (humid, sloping pastures, accumulated dead leaves, presence of waterway) may increase the risk of AM. Specific interventions based on these factors might help to reduce the incidence of AM. PMID- 18396431 TI - Observer-selective double electron-electron-spin resonance, a pulse sequence to improve orientation selection. AB - By pulsed double electron-electron resonance (DEER), distances between spin labels in disordered systems up to 8 nm can be measured. In addition, the relative orientation of the interacting radicals can be determined, provided that the bandwidth of the pulses is sufficiently small. On the other hand, the bandwidth has to exceed the dipolar interaction considerably, because otherwise the DEER modulations become distorted and the modulation depth decreases, making distance determination impossible. Therefore, small bandwidths, i.e. long pulses, place a lower limit on the distance that can be determined. Two new pulse sequences, observer-selective DEER (os-DEER) and dead-time free os-DEER, are introduced that make it possible to use long observer pulses with bandwidths that are smaller than the dipolar interaction. The new pulse sequences do not suffer from the distortions caused by the limited bandwidth of the observer pulses, as demonstrated by measurements on a nitroxide biradical. With observer pulses of 140 ns, i.e., significantly longer than the 32 ns used in the conventional DEER sequence, a dipolar interaction of 7.8 MHz has been measured. PMID- 18396432 TI - Expression of the oligopeptide transporter, PepT1, in larval Atlantic cod (Gadus morhua). AB - The intestinal absorption of di- and tri-peptides generally occurs via the oligopeptide transporter, PepT1. This study evaluates the expression of PepT1 in larval Atlantic cod (Gadus morhua) during the three weeks following the onset of exogenous feeding. Larval Atlantic cod were fed either wild captured zooplankton or enriched rotifers. cDNA was prepared from whole cod larvae preceding first feeding and at 1000 each Tuesday and Thursday for the following three weeks. Spatial and temporal expression patterns of PepT1 mRNA were compared between fish consuming the two prey types using in situ hybridization and quantitative real time PCR. Results indicated that PepT1 mRNA was expressed prior to the onset of exogenous feeding. In addition, PepT1 was expressed throughout the digestive system except the esophagus and sphincter regions. Expression slightly increased following first-feeding and continued to increase throughout the study for larvae feeding on both prey types. When comparing PepT1 expression in larvae larger than 0.15-mg dry mass with expression levels in larvae prior to feeding, no differences were detected for larvae fed rotifers, but the larvae fed zooplankton had significantly greater PepT1 expression at the larger size. In addition, PepT1 expression in the zooplankton fed larvae larger than 0.15-mg dry mass had significantly greater expression than rotifer fed larvae of a similar weight. Switching prey types did not affect PepT1 expression. These results indicate that Atlantic cod PepT1 expression was slightly different relative to dietary treatment during the three weeks following first-feeding. In addition, PepT1 may play an important role in the larval nutrition since it is widely expressed in the digestive tract. PMID- 18396433 TI - Radiation dose to patients and staff during angiography of the lower limbs. Derivation of local dose reference levels. AB - BACKGROUND: The Euratom directive 97/43 recommends the use of patient dose surveys in diagnostic radiology and the establishment of reference dose levels (DRLs). PURPOSE: To perform measurements of the dose delivered during diagnostic angiography of the lower limbs using thermoluminescence dosimeters (TLDs), extraction of DRLs and estimation of the effective dose and radiation risk for this particular examination. METHODS: Dose measurement was performed on 30 patients by using TLD sachets attached in 5 different positions not only on the patient, but also to the radiologist. All the appropriate factors were recorded. Measurement of the ESD was performed after each examination. RESULTS: The mean entrance skin dose (ESD) was calculated to be 70.8, 67.7, 24.3, 18.4, 9.7 mGy at the level of aorta bifurcation, pelvis, femur, knees, and at feet, respectively. The average effective dose is 9.8 mSv with the radiation risks for fatal cancer to be 5.4 x 10(-4). The effective dose of the radiologist was calculated to be 0.023 mSv per procedure. CONCLUSION: Radiation dose variation depends on the physical characteristics of the patient, on the procedure preferences by radiologists and the difficulties in conducting procedures. The main reason for the increased patient dose, compared to other studies, is the number of frames rather than the duration of fluoroscopy. For DSA of the lower limbs, the DRL was chosen to be an entrance skin dose of 96.4 mGy in the pelvic region. The dose to the radiologist is negligible. PMID- 18396434 TI - Transient inhibition of poly(ADP-ribose) polymerase expression and activity by Toxoplasma gondii is dispensable for parasite-mediated blockade of host cell apoptosis and intracellular parasite replication. AB - Poly(ADP-ribose) polymerase-1 (PARP-1; EC 2.4.2.30) is an abundant nuclear protein that is involved in DNA repair, cell cycle control, programmed cell death and transcriptional regulation. It also plays critical roles in the pathogenesis of inflammatory disorders. Here we have performed a detailed analysis of the interplay between the apicomplexan parasite Toxoplasma gondii and host cell PARP and its consequences for the host-parasite relationship. Our results have shown that T. gondii significantly decreased PARP expression in its host cells within 10min of infection but that the amount of PARP normalized during prolonged infection. Importantly, down-regulation of PARP expression after infection abrogated the ADP-ribosylation of acceptor proteins in response to oxidative stress. Overexpression of PARP in RAW264.7 cells revealed that elevated amounts of PARP neither affected host cell invasion nor intracellular development of T. gondii in non-stimulated or IFN-gamma/LPS-stimulated monocytes/macrophages. Furthermore, measurements of the activities of effector caspases 3 and 7 indicated that the blockade of host cell apoptosis by T. gondii occurs independently of the inhibition of PARP after infection. These findings suggest that the prominent decrease of host cell PARP and poly(ADP-ribos)ylation after parasitic infection do not affect the intracellular development of T. gondii in vitro. PMID- 18396435 TI - Infection of the tracheal epithelium by infectious bronchitis virus is sialic acid dependent. AB - Avian Infectious bronchitis virus (IBV) is a coronavirus that infects chickens via the respiratory epithelium as primary target cells. The binding of coronaviruses to the cell surface is mediated by the viral surface protein S. Recently we demonstrated that alpha2,3-linked sialic acid serves as a receptor determinant for IBV on Vero cells and primary chicken embryo kidney cells. Here we analyze the importance of the sialic acid binding activity for the infection of tracheal organ cultures (TOCs) by different IBV strains. Our results show that alpha2,3-linked sialic acid also serves as a receptor determinant on chicken TOCs. Infection of TOCs by IBV results in ciliostasis. Desialylation induced by neuraminidase treatment of tracheal organ cultures prior to infection by IBV delayed the ciliostatic effect or resulted in partial loss of ciliary activity. This effect was observed with both respiratory and nephropathogenic strains. Inhibition of ciliostasis was also observed when TOCs were pretreated with an alpha2,3-specific neuraminidase. Analysis of the tracheal epithelium for reactivity with lectins revealed that the susceptible cells in the epithelium abundantly express alpha2,3-linked sialic acid. These results indicate that alpha2,3-linked sialic acid plays an important role for infection of the respiratory epithelium by IBV. PMID- 18396436 TI - Mannose-binding lectin variant associated with severe malaria in young African children. AB - Mannose-binding lectin (MBL) is a serum protein which initiates innate immune responses to microbial pathogens by binding to non-self surface oligosaccharides. MBL deficiency is the most common congenital immunodeficiency of human and has been shown to predispose to infections, particularly in children and immune compromised. In a matched case-control study among 870 Ghanaian children, we examined the influence of six polymorphisms of the MBL2 gene on Plasmodium falciparum infection and severe malaria. A missense mutation resulting in low MBL activity (MBL2*C) was found in 35% of healthy controls, but in 42% of asymptomatically infected children (P=0.01), and in 46% of patients with severe malaria (P=0.007). Heterozygosity for MBL2*C was associated with increased odds of infection (odds ratio (OR), 1.6; 95% confidence interval (CI), 1.1-2.1), severe malaria (OR, 1.7; 95% CI, 1.2-2.4), and of severe anemia in particular (OR, 2.3; 95% CI, 1.4-3.8). The population attributable fraction of severe malaria cases attributable to MBL2*C heterozygosity was 17%. Our results suggest that the MBL pathway of the complement system is a critical determinant of both, susceptibility to P. falciparum infection and manifestation of severe malaria, particularly in young children in whom specific immune responses are weak or absent. PMID- 18396437 TI - Comparison of Chlamydia trachomatis serovar L2 growth in polarized genital epithelial cells grown in three-dimensional culture with non-polarized cells. AB - A common model for studying Chlamydia trachomatis and growing chlamydial stocks uses Lymphogranuloma venereum serovar L2 and non-polarized HeLa cells. However, recent publications indicate that the growth rate and progeny yields can vary considerably for a particular strain depending on the cell line/type used, and seem to be partially related to cell tropism. In the present study, the growth of invasive serovar L2 was compared in endometrial HEC-1B and endocervical HeLa cells polarized on collagen-coated microcarrier beads, as well as in HeLa cells grown in tissue culture flasks. Microscopy analysis revealed no difference in chlamydial attachment/entry patterns or in inclusion development throughout the developmental cycle between cell lines. Very comparable growth curves in both cell lines were also found using real-time PCR analysis, with increases in chlamydial DNA content of 400-500-fold between 2 and 36 h post-inoculation. Similar progeny yields with comparable infectivity were recovered from HEC-1B and HeLa cell bead cultures, and no difference in chlamydial growth was found in polarized vs. non-polarized HeLa cells. In conclusion, unlike other C. trachomatis strains such as urogenital serovar E, invasive serovar L2 grows equally well in physiologically different endometrial and endocervical environments, regardless of the host cell polarization state. PMID- 18396438 TI - Spectroscopic investigations of carious tooth decay. AB - We report on the elemental composition of healthy and infected part of human tooth using laser induced breakdown spectroscopy (LIBS). We have used prominent constituent transitions in laser-excited tooth to diagnose the state of the tooth. A nanosecond laser pulse (355nm, 5ns) was used as an ablating pulse and the sodium (3s2S-3p2P) at 588.99 and (3s2S-3p2P) at 589.99nm, strontium (5s21S 1s5P) at 460.55nm, and calcium (3d3D-4f 3F0) at 452.55nm transitions for spectroscopic analysis. The spectroscopic observations in conjunction with discriminate analysis showed that calcium attached to the hydroxyapatite structure of the tooth was affected severely at the infected part of the tooth. The position-time plots generated from two-dimensional (2D) images conclusively showed a decrease in calcium concentration in the infected region of the irradiated tooth. Using the technique, we could distinguish between the healthy and carious parts of the tooth with significant accuracy. PMID- 18396439 TI - A therapeutic taskscape: theorizing place-making, discipline and care at a camp for troubled youth. AB - Ethnographic research at a therapeutic camping program for troubled youth in Vermont suggests that therapeutic landscapes can be productively theorized as 'taskscapes' [Ingold, T., 1993. The temporality of the landscape. World Archaeology 25(2), 152-174]. Rather than searching for the elements of material landscape that make a particular place health-giving, the taskscape perspective focuses on the on-going activities that produce distinctive material landscapes, discipline, individual meaning, and therapeutic experience. This paper engages the debate over analyzing health care institutions as spaces of control or care, finding that both perspectives can be accommodated within the taskscape approach. PMID- 18396440 TI - Prolylcarboxypeptidase: a cardioprotective enzyme. AB - Prolylcarboxypeptidase (PRCP) is involved in regulating the blood flow through active tissues in order to preserve the internal environment. The expression of PRCP in tissues is determined by a number of pharmacological stimuli such as glucocorticoids and a combination of dexamethasone plus the mu-opioid receptor agonist [D-Ala2, N-Me-Phe4, Gly5-ol]-Enkephalin acetate. PRCP is an enzyme which is associated with preeclampsia, rheumatoid arthritis, and tonsillitis. The interplay between inward cellular signalling required for induced and basal transcription, and PRCP expression have not been mechanistically characterized. Molecules modulated by PRCP include angiotensin II (Ang II), angiotensin III (Ang III), alpha-MSH, and prekallikrein (PK), demonstrating its cardiovascular protective role. In addition to regulating vascular tone, PRCP may modulate proliferation, cell migration, and angiogenesis through regulating angiotensin molecules--and bradykinin--induced endothelium activation. The anti-hypertensive and proinflammatory properties of PRCP implicate that this enzyme may well be an accessible target for anti-inflammatory therapy. PMID- 18396441 TI - Regions, systems, and the brain: hierarchical measures of functional integration in fMRI. AB - In neuroscience, the notion has emerged that the brain abides by two principles: segregation and integration. Segregation into functionally specialized systems and integration of information flow across systems are basic principles that are thought to shape the functional architecture of the brain. A measure called integration, originating from information theory and derived from mutual information, has been proposed to characterize the global integrative state of a network. In this paper, we show that integration can be applied in a hierarchical fashion to quantify functional interactions between compound systems, each system being composed of several regions. We apply this method to fMRI datasets from patients with low-grade glioma and show how it can efficiently extract information related to both intra- and interhemispheric reorganization induced by lesional brain plasticity. PMID- 18396442 TI - Adenoid cystic carcinoma of intraoral minor salivary glands. AB - We present a retrospective multicentric study of 67 patients with adenoid cystic carcinomas of the intraoral minor salivary glands, who were diagnosed and treated between 1986 and 2004 in the Operative Units of Maxillofacial Surgery of the University of Parma and of the Bellaria Hospital in Bologna. There were 28 males and 39 females with a mean age of 54.5 years (range 28-84 years). The palate was the most common site of involvement, with 71.6% of cases. All the patients were treated with surgery as the primary modality. Neck dissection was performed in 9% of the patients, and more than half (59.7%) were treated with adjuvant radiation therapy. All surviving patients had a minimum of 2 years follow-up, with a range of 2-20 years (median: 12 years). Of the 67 patients examined in this study, 19 were dead at the end of the follow-up, 16 of them due to tumor disease. Distant or loco-regional recurrence was documented in 29 of the 67 patients (43.3%). Disease-free intervals ranged from 1 month to 12 years. Twenty-two percent of patients had loco-regional failure, whereas 29.8% had distant metastases. The disease specific survival rates were 76.1% at 5 years and 74% at 10 and 15 years.The actuarial loco-regional control rates at 5-, 10- and 15-years were 79.7%, 74.8% and 70%, respectively. The actuarial freedom from distant relapse was 76.5% at 5 years, 64.9% at 10 years and 61.1% at 15 years. Stage T, cervical lymph node metastasis and surgical margin status proved to make a statistically significant contribution when describing the outcome. PMID- 18396443 TI - Collagen telopeptide (serum CTX) and its relationship with the size and number of lesions in osteonecrosis of the jaws in cancer patients on intravenous bisphosphonates. AB - Osteonecrosis of the jaws (ONJ) is an important possible late adverse effect of bisphosphonates. Serum C-terminal cross-linking telopeptide of type I collagen (CTX) can determine bone turnover and can act as a biological marker of bone resorption. We studied this biological marker in 15 patients (Group 1) with bisphosphonate-induced ONJ comparing with a control group of 10 healthy people matched by age and gender. We found no statistically significant relationships in Group 1 either between the serum CTX and the number of areas of exposed bone nor with the size of the osteonecrotic area. PMID- 18396444 TI - The prognostic significance of tumor vascular invasion and its association with plasma Epstein-Barr virus DNA, tumor volume and metabolic activity in locoregionally advanced nasopharyngeal carcinoma. AB - Parapharyngeal tumor invasion is a known predictor of distant recurrence in stage II-III nasopharyngeal carcinoma (NPC). This study evaluated the prognostic significance of parapharyngeal and cavernous sinus vascular invasion in stage III IV NPC, and its association with plasma Epstein-Barr virus (pEBV) DNA, disease stage, tumor volume and metabolic activity. Eligible patients underwent magnetic resonance imaging (MRI), 18F-fluorodeoxyglucose (FDG)-positron-emission tomography (PET) and blood sampling for pEBV DNA before undergoing concurrent chemoradiotherapy. Relationship between treatment outcome and tumor vascular invasion was analyzed using Cox regression. Logistic regression was used to analyze the association between vascular invasion and other cofactors. Fifty seven patients with predominantly T3-T4 or N2-N3 stage disease were enrolled. Parapharyngeal invasion was present in 56% and cavernous sinus invasion in 19% of tumors. Multivariate analysis showed that tumor vascular invasion did not predict treatment outcome, while tumor FDG-uptake was the only significant factor that predicted survival and recurrence. Tumor vascular invasion was associated with T stage, but not pEBV DNA or tumor volume. Parapharyngeal and cavernous sinus invasion were not significant predictors of distant recurrence following chemoradiotherapy in our cohort with locoregionally advanced NPC. PMID- 18396446 TI - Primary mucosal melanoma of the head and neck. Comparison of clinical presentation and histopathologic features of oral and sinonasal melanoma. AB - We reviewed all cases of head and neck mucosal melanomas (HNMM) treated at Emory University and affiliated hospitals during a 20 year period and evaluate overall survival, recurrences and efficacy of treatment. Comparisons were made between sinonasal melanoma (SNM) and oral cavity melanoma (OCM) including clinical features, histopathologic features, treatment, and clinical outcomes. We analyzed pathologic features and clinical outcomes of 22 cases of primary SNM and eight cases of OCM treated at Emory University Hospital between 1986 and 2006. Sixteen patients were stage I (53%), nine patients were stage II (30%) and five patients were stage III (17%). Mean age was 67.5 years with a range from 32 to 85 years. Sixty percent were men and 92% were white. The average follow-up time was 22.1 months and the median was 15.5 months. The average delay between onset of symptoms and diagnosis was 5.6 months. Patients with SNM had a lower incidence of nodal metastasis at initial presentation when compared with OCM (22% vs. 50%) (p<.08). Surgical resection was the primary treatment and was performed in 27/30 patients (90%). Two patients with SNM and 1 patient with OCM, because of advanced stage at diagnosis had no treatment. Radiation was used in 50% (15) of the patients and radiation as postoperative therapy was administered to 44.8% (13) of the patients. The combination of surgery, radiation and adjuvant therapy was administered to 39.2% (11/2 8) of the patients. The recurrence rate at the time of closing this study was 43.3% (13 patients) with a mean time for development of recurrences of 13.1 months. Twenty-five patients (83%) died during the course of this study. Nineteen deaths (82%) corresponded to patients with SNM and six deaths (75%) to patients with OCM. The overall survival mean time was 21 months (median 12.5) with a range between 1 and 143 months. The 1, 2, 3, and 5 year survival rate for this study was 53.3%, 40%, 15% and 10%, respectively. The oral cavity was the location for the two patients still alive after 5 years. Although survival time correlated with Stage, particularly Stage 1, this was not statistically significant. Survival time did not correlate with surgery or adjuvant therapy. Statistically significant differences were noted between the pathologic features of OCM and SNM. These significant pathologic differences did not correlate with disease specific survival: OCM and SNM (median survival, 17 months vs. 12 months). Mucosal melanoma of the head and neck is a rare entity. Unfortunately, most patients present with advanced local disease. Local, regional recurrences and distant metastasis still occur despite the implementation of aggressive treatment, including surgery, radiation and adjuvant therapy. Despite significant pathologic differences between SNM and OCM, no survival advantage was seen. PMID- 18396445 TI - In vivo imaging of oral neoplasia using a miniaturized fiber optic confocal reflectance microscope. AB - The purpose of this study was to determine whether in vivo images of oral mucosa obtained with a fiber optic confocal reflectance microscope could be used to differentiate normal and neoplastic tissues. We imaged 20 oral sites in eight patients undergoing surgery for squamous cell carcinoma. Normal and abnormal areas within the oral cavity were identified clinically, and real-time videos of each site were obtained in vivo using a fiber optic confocal reflectance microscope. Following imaging, each site was biopsied and submitted for histopathologic examination. We identified distinct features, such as nuclear irregularity and spacing, which can be used to qualitatively differentiate between normal and abnormal tissue. Representative confocal images of normal, pre neoplastic, and neoplastic oral tissue are presented. Previous work using much larger microscopes has demonstrated the ability of confocal reflectance microscopy to image cellular and tissue architecture in situ. New advances in technology have enabled miniaturization of imaging systems for in vivo use. PMID- 18396447 TI - Potential role of mast cells in hamster cheek pouch carcinogenesis. AB - During the process of activation, mast cells release products stored in their granules. Tryptase, a protease released from mast cell granules after activation, induces tumor cell proliferation through the activation of PAR-2 (protease activated receptor 2) on the plasma membrane of carcinoma cells. Chemical cancerization (DMBA) of the hamster cheek pouch is the most accepted model of oral cancer. However, there are no reports on the activation of mast cells during experimental carcinogenesis or on the correlation between mast cell activation and cell proliferation. The aim of the present study was to evaluate the potential effect of mast cells on the proliferation of epithelial cells at different times during the cancerization process. Paraffin serial sections of cancerized, tumor-bearing pouches were stained with Alcian Blue-Safranin to identify the different degrees of mast cell activation. Immunohistochemistry was performed to identify BrdU-positive cells to study tumor cell proliferation. Mast cells were counted and grouped into two categories: inactive mast cells AB-S+++ (red) and active mast cells AB+++S- (blue). Mast cell counts were performed in tumor stroma, base of the tumor (connective tissue immediately below the exophytic tumor), connective and muscle tissue underlying the cancerized epithelium (pouch wall) and adventitious tissue underlying the pouch wall. There was a significant increase in the number of mast cells at the base of tumors (p<0.001) compared to the number of mast cells in the wall of the pouch and in tumor stroma. In normal non-cancerized pouches, inactive mast cells were prevalent both in the wall (AB:S=1:2.15; p<0.001) and in the adventitious tissue (AB:S=1:1.6; p<0.004) of the hamster cheek pouch. At most of the experimental times examined, the ratio of active/inactive mast cells (AB/S) in the wall approximated unity and even reverted. The ratio of mast cells was AB:S 1:1.05 at the base of the tumor and 1:0.24 in tumor stroma (p<0.001). The evaluation of epithelial nuclei labeled for BrdU revealed a statistically significant increase in cells undergoing DNA synthesis in the epithelium of the wall of the cancerized pouch compared to control (p<0.017). Tumor parenchyma exhibited a highly statistically significant increase in DNA synthesis compared to control (p<0.001) and compared to the epithelium of the wall of the cancerized pouch (p<0.036). We conclude that mast cell activation in this model is associated to the increase in tumor cell proliferation, conceivably mediated by the release of tryptase. PMID- 18396448 TI - Clocks and switches: bacterial gene regulation by DNA adenine methylation. AB - N(6) methylation in adenosine moieties causes changes in DNA structure and can modulate DNA-protein interactions. In both alpha-Proteobacteria and gamma Proteobacteria, postreplicative formation of N(6)-methyl-adenine regulates transcription of specific genes and provides two general types of controls: (i) clock-like controls that permit transient gene transcription during a specific stage of DNA replication; (ii) switch-like controls in which transcription is regulated by a DNA methylation pattern. DNA adenine methylation may also regulate gene expression by affecting nucleoid topology. Recent transcriptomic studies have unveiled novel cases of genes regulated by DNA adenine methylation, including virulence genes of bacterial pathogens. PMID- 18396449 TI - Control of gene expression by type III secretory activity. AB - The bacterial flagellum and the highly related injectisome (or needle complex) are among the most complicated multi-protein structures found in Gram-negative microorganisms. The assembly of both structures is dependent upon a type III secretion system. An interesting regulatory feature unique to these systems is the coordination of gene expression with type III secretory activity. This means of regulation ensures that secretion substrates are expressed only when required during the assembly process or upon completion of the fully functional structure. Prominent within the regulatory scheme are secreted proteins and type III secretion chaperones that exert effects on gene expression at the transcriptional and post-transcriptional levels. Although the major structural components of the flagellum and injectisome systems are highly conserved, recent studies reveal diversity in the mechanisms used by secretion substrates and chaperones to control gene expression. PMID- 18396450 TI - Nutritional control via Tor signaling in Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae senses and responds to nutrients by adapting its growth rate and undergoing morphogenic transitions to ensure survival. The Tor pathway is a major integrator of nutrient-derived signals that in coordination with other signaling pathways orchestrates cell growth. Recent advances have identified novel Tor kinase substrates and established the protein trafficking membranous network and the nucleus as platforms for Tor signaling. These and other recent findings delineate distinct signaling branches emanating from membrane-associated Tor complexes to control cell growth. PMID- 18396451 TI - Theoretical investigation of the molecular, electronic structures and vibrational spectra of a series of first transition metal phthalocyanines by Z. Liu et al. AB - A recent paper by Lui et al. [Z. Liu, X. Zhang, Y. Zhang, J. Jiang, Spectrochim. Acta A 67 (2007) 1232] reported on the theoretical investigations of the fully optimized geometries and electronic structures of iron (II) phthalocyanine (FePc) with the singlet spin state carried out with the restricted density functional theory (DFT) method, where the B3LYP functional was adopted for the exchange correlation term; however, the triplet spin state was experimentally reported, and we also obtained the triplet spin state by the unrestricted DFT calculations. PMID- 18396452 TI - pH-dependent Raman study of pyrrole and its vibrational analysis using DFT calculations. AB - Raman spectra of pyrrole in aqueous medium at different pH values, 2.5, 5.5, 7.5 and 10.5 were recorded in the two spectral regions, 1,040-1,160 cm(-1) and 3,300 3,360 cm(-1) and pH dependence of the linewidth, peak position and intensity of the Raman bands corresponding to the ring breathing and symmetric nu(N-H) stretching modes were examined. A linear pH dependence of the peak positions for the ring breathing mode and a maximum at nearly neutral pH (7.5) for the symmetric nu(N-H) normal mode is observed, whereas the linewidth (FWHM) shows almost no variation with the change of pH. A slight decrease in the wavenumber position of the nu(N-H) mode at pH value >7.5 indicates that the influence of deprotonation is small, which results from a weak interaction between the reference molecule and the surrounding environment. The density functional theory (DFT) calculations were made primarily to obtain the optimized geometry and vibrational spectra of pyrrole in the ground electronic state using B3LYP functional and the highest level basis set 6-311++G(d,p). The assignments of the normal modes of pyrrole were made on the basis of potential energy distribution (PED). The calculations were also performed on protonated and deprotonated structures of pyrrole. PMID- 18396453 TI - Effect of motion imagery to counter rest-induced suppression of F-wave as a measure of anterior horn cell excitability. AB - OBJECTIVE: To test if motor imagery prevents the rest-induced suppression of anterior horn cell excitability. METHODS: Ten healthy subjects underwent two separate experiments, each consisting of stimulating the median nerve 100 times and recording F-waves from abductor pollicis brevis (APB) in three consecutive sessions: (1) after muscle exercise to standardize the baseline, (2) after immobilization of APB for 3h and (3) after muscle exercise to check recovery. We instructed the subject to volitionally relax APB in experiment 1 (relaxation task), and to periodically simulate thumb abduction without actual movement in experiment 2 (imagery task). RESULTS: F-wave persistence and amplitude declined after relaxation task and recovered quickly after exercise, but changed little with imagery task. F-wave latencies showed no change when analyzed individually. The frequency distribution of collective F-waves recorded from all subjects remained the same after relaxation task, but showed a shift toward longer latencies after imagery task. CONCLUSIONS: Mental imagery without overt motor output suffices to counter the effect of sustained volitional muscle relaxation, which would, otherwise, cause a reversible reduction in anterior horn cell excitability. SIGNIFICANCE: This finding documents the importance of central drive for spinal excitability, which affects F-wave studies of a paretic muscle. PMID- 18396454 TI - Abnormal EEG complexity in patients with schizophrenia and depression. AB - OBJECTIVE: Schizophrenics are usually unable to perform well on cognitive tasks due to disturbances in cortical information processing that are observable as abnormalities in electroencephalogram (EEG) signals. However, whether such cortical disturbances can be assessed by quantitative EEG analysis remains unclear. The purpose of this study was to characterize EEG disturbances, using the Lempel-Ziv complexity (LZC), in the subjects with schizophrenia at rest or while performing mental arithmetic tasks. The results were compared to those from the subjects with depression and with healthy controls. METHODS: The subjects included 62 schizophrenia patients, 48 depression patients and 26 age-matched healthy controls. EEG was recorded under two conditions: (i) resting with eyes closed, and (ii) a mentally active condition wherein the subjects were asked to subtract 7 from 100 iteratively with their eyes closed. EEG signals were analyzed by LZC and conventional spectral methods. RESULTS: In all the groups, LZC of EEG decreased during the mental arithmetic compared with those under the resting conditions. Both the schizophrenia and the depression groups had a higher LZC (p<0.05) than the controls. Also, the schizophrenia group had a lower LZC (p<0.05) than the depression group during the mental arithmetic task as well as during the resting state. Significant differences in LZC, at some symmetrically located loci (FP1/FP2, F7/F8), between the two hemispheres were found in all the patient groups only during the arithmetic task. CONCLUSIONS: Compared with conventional spectral analysis, LZC was more sensitive to both the power spectrum and the temporal amplitude distribution. LZC was associated with the ability to attend to the task and adapt the information processing system to the cognitive challenge. Thus, it would be useful in studying the disturbances in the cortical information processing patients with depression or schizophrenia. SIGNIFICANCE: LZC of EEG is associated with mental activity. Thus, LZC analysis can be an important tool in understanding the pathophysiology of schizophrenia and depression in future studies. PMID- 18396455 TI - Zonisamide decreases cortical excitability in patients with idiopathic generalized epilepsy. AB - OBJECTIVE: To evaluate changes in cortical excitability after long-term zonisamide (ZNS) administration. METHODS: Fifteen drug-naive idiopathic generalized epilepsy (IGE) patients (8 male, mean age 24.9 years) were enrolled. The transcranial magnetic stimulation (TMS) parameters obtained using two Magstim 200 stimulators were resting motor threshold (RMT), motor evoked potential (MEP) amplitudes, cortical silent period (CSP), intracortical inhibition (ICI), and intracortical facilitation (ICF). TMS parameters were compared before and after ZNS administration. RESULTS: All patients were administered ZNS monotherapy (200 mg/day) for 8 weeks. No patient reported seizures during the study period. After ZNS treatment MEP amplitudes were significantly reduced in right (-34.2%) and left hemispheres (-37.0%) (Wilcoxon's signed rank test after Bonferroni's correction for multiple comparisons, P < 0.05). Mean RMT, CSP, and ICI/ICF were not changed by ZNS (P > 0.05). CONCLUSIONS: These findings suggest that ZNS decreases cortical excitability in patients with IGE and a MEP amplitude is a useful TMS parameter for evaluating changes in cortical excitability induced by ZNS. SIGNIFICANCE: The findings in this study are helpful to understand how ZNS affects the excitability of the motor cortex in patients with IGE. PMID- 18396456 TI - NT-proBNP response to dobutamine stress echocardiography predicts left ventricular contractile reserve in dilated cardiomyopathy. AB - BACKGROUND: Brain natriuretic peptide (BNP) and left ventricular (LV) inotropic reserve are major prognostic indexes in heart failure (HF). AIMS: To investigate the relationship between N-terminal-proBNP (NT-proBNP) changes in response to dobutamine stress echocardiography (DSE) and the LV inotropic reserve, in HF patients with dilated cardiomyopathy (DC). METHODS: We studied 41 patients with DC, LVEF 31.6+/-7.7%, NYHA class II-III and 15 controls. Plasma NT-proBNP levels were measured before and 60 min after three 5-min stages of dobutamine (5 to 15 microg/kg/min). RESULTS: Based on NT-proBNP changes in response to dobutamine, patients were categorized into two groups: In Group A circulating NT-proBNP levels fell (-16.6+/-7.8%), and in Group B they increased (8.4+/-9.1%). Group A had a marked improvement in WMSI compared to Group B (32.1+/-9.7% vs. 18.8+/ 15.9%, p<0.001). Multivariate analysis showed that NT-proBNP changes were an independent predictor of LV inotropic reserve (b= -0.55, p<0.001). A reduction of 21.3% in plasma NT-proBNP levels in response to dobutamine predicted an improvement in WMSI of >25% with a sensitivity of 100% and a specificity of 92.3%. CONCLUSIONS: NT-proBNP changes in response to dobutamine reflect improvement in LV contractility and constitute an independent predictor of LV inotropic reserve in patients with DC. PMID- 18396457 TI - Status dissociatus evolving from REM sleep behaviour disorder in multiple system atrophy. AB - OBJECTIVE: We present two patients, a 52-year-old man and a 56-year-old woman, with rapid eye movement sleep behaviour disorder (RBD) since the age of 50. METHOD: In both the patients RBD was videopolysomnographically documented. RESULTS: Both patients developed, with time, autonomic and motor symptoms consistent with the diagnosis of multiple system atrophy. During the course of the disease RBD episodes diminished in frequency but the patients' sleep became even more abnormal, with nearly continuous motor and verbal abnormal behaviours and ambiguous and rapid oscillations of state-determining polysomnographic variables, now consistent with status dissociatus (SD). CONCLUSION: If SD represents the evolution and most extreme form of RBD it should be investigated longitudinally in neurodegenerative diseases. PMID- 18396458 TI - Mesenchymal stem cells: isolation, characterisation and in vivo fluorescent dye tracking. AB - Cell therapies have been used to regenerate the heart by direct myocardial delivery, by coronary infusion and by surface attached scaffolds. Multipotent mesenchymal stem cells (MSC) with capacity to differentiate into cardiomyocytes and other cell lines have been predominantly trialled in rodents. However, large animal models are increasingly needed to translate basic research into new, safe regenerative therapies. Understanding the mode of action of cell therapies in the mammalian heart has been limited by cell tracking capability. This study examined the ability to track the fate of allogeneic MSC in sheep using various fluorescent dyes. MSC isolated from sheep bone marrow were grown in culture following extraction and flow cytometric characterisation. After labelling with fluorescent tracking dyes (e.g. CFSE and DiI) cells were tested for in vitro and in vivo signal up to six weeks. Labelling effect on cell division and differentiation was studied. Several dyes lost fluorescence and slowed cell division. However, the thiol reactive dye CM-DiI showed detectable in vivo fluorescence in labelled MSC six weeks after injection into sheep skeletal muscle and two weeks after implantation of an MSC coated biomaterial scaffold. CM-DiI labelled MSC differentiated in vitro showed label retention over four weeks. The fluorescent membrane dye CM-DiI tracks implanted sheep MSC and provides an alternative to traditional cell markers such as gene modified GFP. PMID- 18396459 TI - Exercise-induced right ventricular dysplasia/cardiomyopathy--an emerging condition distinct from arrhythmogenic right ventricular dysplasia/cardiomyopathy. AB - We report a case of a 32-year-old female world champion triathlete who developed exercise induced recurrent ventricular tachycardia (VT). Investigations supported a diagnosis of the newly recognised condition "exercise induced right ventricular dysplasia/cardiomyopathy" (EIRVD/C). The VT could be easily terminated by burst pacing from the RV apex and the athlete has been successfully treated with an internal cardioverter defibrillator (ICD). PMID- 18396460 TI - Giant congenital coronary artery aneurysm in pulmonary atresia with intact septum. AB - Congenital coronary artery fistula is frequently associated with pulmonary atresia and intact ventricular septum (PA/IVS). Aneurysmal dilatation is rare, and can be complicated by rupture, distal thrombi and myocardial ischaemia. PMID- 18396461 TI - The fine structure of the midgut in the mite Anystis baccarum (L.) (Acari, Actinedida: Anystidae). AB - The ventriculus and the midgut caeca of the fed females of Anystis baccarum (L.) were investigated by using light and electron microscopy. In addition to the main type of polyfunctional digestive cells, special secretory cells were detected in the anterior region of the ventriculus. The shape and the ultrastructure of the digestive cells vary depending on their physiological state. Intracellular digestion, absorption or excretion processes prevail at different stages of the cell cycle. The secretory cells are characterized by the presence of extensive rough endoplasmic reticulum, filling whole space of the cell. These cells do not contain the apical network of pinocytotic canals, which are typical for the digestive cells. Three types of secretory granules were found in the cytoplasm of the secretory cells that probably correspond to three sequential stages of granulogenesis. The primary secretory granules are formed by the fusion of Golgi vesicles. The primary granules fuse to form complex vesicles with heterogeneous contents. These secondary granules aggregate to form very large inclusions of high electron density (tertiary secretory granules), which probably represent the storage of the secretory product. All types of secretory granules were observed close to the apical plasmalemma. PMID- 18396462 TI - Structure of ovaries and oogenesis in dermapterans. I. Origin and functioning of the ovarian follicles. AB - The ovaries of the studied earwig species (Forficula auricularia, Chelidurella acanthopygia, Doru lineare and Opisthocosmia silvestris) are meroistic polytrophic and composed of numerous short ovarioles that consist of a terminal filament, germarium and vitellarium. The germaria of adult females comprise meiotic (pachytene) and postmeiotic (differentiating) germ cell clusters, as well as small prefollicular cells. All germ cell clusters consist of two cells that are connected by a single intercellular bridge. In the vitellarium there are usually 2 ovarian follicles only. The individual follicle consists of a transcriptionally dormant oocyte and a single, polyploid nurse cell and is surrounded by a layer of somatic follicular cells (FCs). During previtellogenesis the nurse cell enlarges and becomes highly transcriptionally active. Concurrently its nucleus attains a characteristic, irregular shape. In the nurse cell nucleus of one studied species, F. auricularia, in addition to chromatin aggregations and RNA- and Ag-NOR-positive nucleoli, a single compact DNA-positive body is present. During advanced vitellogenesis the molecules synthesized in the nurse cells (RNAs, proteins, as well as nurse cell organelles) are transferred to the ooplasm via the intercellular bridge. During this transfer the nurse cell nucleus is retained in the cell centre and does not occlude the intercellular bridge. The results of our studies indicate that such position of the nurse cell nucleus is maintained solely by its extended shape. In other words, the rigid extensions keep the nucleus in the cell centre while the cytoplasm flows, in between these extensions, towards the intercellular bridge connecting the nurse cell with the oocyte. PMID- 18396463 TI - A tale of two citizens: sleeping with a snoring generator. PMID- 18396464 TI - Primary care and emergency physician attitudes and beliefs related to patients with psychogenic nonepileptic spells. AB - Psychogenic nonepileptic spells (PNES) are typically evaluated by primary care and emergency physicians before neurologists. The attitudes and beliefs of such physicians about PNES may impact long-term outcomes and quality of care for these challenging patients. A 21-question survey was created to assess knowledge of, beliefs about, and attitudes toward the diagnosis and management of PNES. The survey found misperceptions that PNES are voluntary, that video/EEG monitoring is not needed to confirm the diagnosis of PNES, and that clinical history is sufficient for diagnosis. Ninety-five percent believe that counseling and psychotherapy are the most appropriate therapies. Ongoing education and outreach to referring physicians about PNES are indicated, especially because early diagnosis of PNES has been associated with better outcomes. PMID- 18396465 TI - Health-related quality of life in epilepsy: findings obtained with a new Italian instrument. AB - The aim of the present study was to evaluate the Epi-QoL, a new Italian-specific measure of health-related quality of life (HRQOL) for adults with epilepsy; the clinical variables that affected the HRQOL score were also assessed. The Epi-QoL is a 46-item self-administered questionnaire focusing on six domains: Physical Functioning, Cognitive Functioning, Emotional Well-Being, Social Functioning, Seizure Worry, and Medication Effects. Eight hundred fifteen patients recruited from 24 secondary and tertiary Italian centers for the care of epilepsy were assessed. The results supported the reliability and validity of the Epi-QoL as a measure of HRQOL. The variables that significantly affected HRQOL scores were: geographic area, gender, seizure frequency, prognostic categories, number of medications, comorbidity, presence of cognitive impairment, psychiatric disturbances, and disability. We believe that a new specific questionnaire for the evaluation of HRQOL in Italy can contribute much to the understanding of the influence of epilepsy on patients' lives. PMID- 18396466 TI - Emetic stimulation inhibits the swallowing reflex in decerebrate rats. AB - The effects of emetic stimulation on the swallowing reflex were investigated in decerebrated rats. Hypoxia, gastric distension and LiCl administration were used as emetic stimulations. The swallowing reflex was elicited by electrical stimulation of the superior laryngeal nerve (SLN, 20 Hz, 3-5 V, 0.3 ms duration) for 20 s. To examine the effect of hypoxia, nitrogen gas was inhaled under artificial ventilation. There were significantly fewer swallows during a decrease in PO(2) than under air ventilation (p<0.05). The number of swallows during 3-ml stomach distension was significantly lower than that before distension (p<0.05). Intravenous administration of LiCl (100 mg/kg) also significantly reduced the number of swallows (p<0.05). The combination of SLN stimulation and emetic stimuli occasionally produced burst activity of abdominal muscles, which might be associated with the gag reflex. Both the gag and swallowing reflexes are well known to be mediated by the nucleus of the solitary tract. The physiological roles of the gag reflex and the swallowing reflex are considered to be reciprocal. Taken together, these results suggest that emetic stimulation inhibits the swallowing pattern generator via the nucleus of the solitary tract, which in turn facilitates the gag reflex. PMID- 18396467 TI - Genetic variation and haplotype structures of innate immunity genes in eastern India. AB - This study reports results of an extensive and comprehensive study of genetic diversity in 12 genes of the innate immune system in a population of eastern India. Genomic variation was assayed in 171 individuals by resequencing approximately 75kb of DNA comprising these genes in each individual. Almost half of the 548 DNA variants discovered was novel. DNA sequence comparisons with human and chimpanzee reference sequences revealed evolutionary features indicative of natural selection operating among individuals, who are residents of an area with a high load of microbial and other pathogens. Significant differences in allele and haplotype frequencies of the study population were observed with the HapMap populations. Gene and haplotype diversities were observed to be high. The genetic positioning of the study population among the HapMap populations based on data of the innate immunity genes substantially differed from what has been observed for Indian populations based on data of other genes. The reported range of variation in SNP density in the human genome is one SNP per 1.19kb (chromosome 22) to one SNP per 2.18kb (chromosome 19). The SNP density in innate immunity genes observed in this study (>3SNPskb(-1)) exceeds the highest density observed for any autosomal chromosome in the human genome. The extensive genomic variation and the distinct haplotype structure of innate immunity genes observed among individuals have possibly resulted from the impact of natural selection. PMID- 18396468 TI - Rad52 sumoylation and its involvement in the efficient induction of homologous recombination. AB - The protein Rad52 is a key player in various types of homologous recombination and is essential to maintenance of genomic integrity. Although evidence indicates that Rad52 is modified by SUMO, the physiological relevance of this sumoylation remains unclear. Here, we identify the conditions under which Rad52 sumoylation is induced, and clarify the role of this modification in homologous recombination. Oligomerization of Rad52 was a prerequisite for sumoylation, and the modification occurred in the cell proceeding S phase being exposed to the DNA damaging agent methyl methanesulfonate (MMS). Following exposure to MMS, sumoylated Rad52 accumulated in rad51 cells, but not in the recombination-related gene mutants, rad54, rad55, rad59, sgs1, or srs2. The accumulation of sumoylated Rad52 was suppressed in rad51 cells expressing Rad51-K191R, an ATPase-defective protein presumed to be recruited to ssDNA. Although the sumoylation defective mutant rad52-3KR (K10R/K11R/K220R) showed no defect in mating-type switching, which did not lead to Rad52 sumoylation in wild-type cells, the mutant did demonstrate a partial defect in MMS-induced interchromosomal homologous recombination. PMID- 18396469 TI - Gas and aerosol mixing in the acinus. AB - This review is concerned with mixing and transport in the human pulmonary acinus. We first examine the current understanding of the anatomy of the acinus and introduce elements of fluid mechanics used to characterize the transport of momentum, gas and aerosol particles. We then review gas transport in more detail and highlight some areas of current research. Next we turn our attention to aerosol transport and in particular to mixing within the alveoli. We examine the factors influencing the level of mixing, review the concept of chaotic convective mixing, and make some brief comments on how mixing affects particle deposition. We end with a few comments on some issues unique to the neonatal and developing lung. PMID- 18396471 TI - Development and validation of a reverse-phase HPLC with fluorescence detector method for simultaneous determination of CZ48 and its active metabolite camptothecin in mouse plasma. AB - A simple and sensitive high-performance liquid chromatography (HPLC) assay for the analysis of CZ48, a potent anticancer candidate, and its active metabolite camptothecin (CPT) in mouse plasma was developed and validated. CZ44 was used as an internal standard (IS). The samples were injected onto a C18 Synergi Polar-RP column (4 microm, 150 mm x 4.60 mm) maintained at 30 degrees C. The identification of peaks showed high specificity. Shimadzu RF-10AXL fluorescence detector was used at the excitation and emission of 380 and 418 nm, respectively. The mean recoveries were 81.41+/-0.035%, 86.00+/-0.053% and 82.21+/-0.020% for CZ48 and 76.01+/-0.028%, 77.04+/-0.042% and 85.93+/-0.023% for CPT at three concentrations of 10, 100 and 900 ng/ml, respectively. The calibration curve was linear (r(2)=0.9999) over CZ48 and CPT concentrations ranging from 5 to 1000 ng/ml and 10-1000 ng/ml (n=6), respectively. The method had an accuracy of >95% and intra- and inter-day precision (RE%) of <1.2% and <2.2% for CZ48 and CPT, respectively, at three different concentrations (10, 100 and 900 ng/ml). The lower limit of quantification (LLOQ) using 0.1 ml mouse plasma was 10 ng/ml for CZ48 and 5 ng/ml for CPT. Stability studies showed that CZ48 and CPT were stable in mouse plasma after 4h incubation at room temperature or after 1 month storage at -80 degrees C with three freeze/thaw cycles. The method reported is simple, reliable, precise and accurate and confirmed by the determination of plasma samples in the mice after oral administration of CZ48. PMID- 18396470 TI - Spinal serotonin receptor activation modulates the exercise ventilatory response with increased dead space in goats. AB - Small increases in respiratory dead space (VD) augment the exercise ventilatory response by a serotonin-dependent mechanism known as short-term modulation (STM). We tested the hypotheses that the relevant serotonin receptors for STM are in the spinal cord, and are of the 5-HT2-receptor subtype. After preparing adult female goats with a mid-thoracic (T6-T8) subarachnoid catheter, ventilation and arterial blood gases were measured at rest and during treadmill exercise (4.8 km/h; 5% grade) with and without an increased VD (0.2-0.3 L). Measurements were made before and after spinal or intravenous administration of a broad-spectrum serotonin receptor antagonist (methysergide, 1-2mg total) and a selective 5-HT2 receptor antagonist (ketanserin, 5-12 mg total). Although spinal methysergide had no effect on the exercise ventilatory response in control conditions, the augmented response with increased VD was impaired, allowing Pa(CO)(2) to increase from rest to exercise. Spinal methysergide diminished both mean inspiratory flow and frequency responses to exercise with increased VD. Spinal ketanserin impaired Pa(CO)(2) regulation with increased VD, although its ventilatory effects were less clear. Intrathecal dye injections indicated CSF drug distribution was caudal to the upper cervical spinal cord and intravenous drugs at the same total dose did not affect STM. We conclude that spinal 5-HT2 receptors modulate the exercise ventilatory response with increased VD in goats. PMID- 18396472 TI - Analysis of amphetamine-type stimulants and their metabolites in plasma, urine and bile by liquid chromatography with a strong cation-exchange column-tandem mass spectrometry. AB - The aim of this work was to develop and validate a method for analysing amphetamine-type stimulants (ATSs) and their metabolites in plasma, urine and bile by liquid chromatography with a strong cation-exchange column-tandem mass spectrometry, and to apply it to the pharmacokinetic study of ATSs. 3,4 Methylenedioxymethamphetamine, methamphetamine, ketamine and their main metabolites, 4-hydroxy-3-methoxymethamphetamine, 3,4-methylenedioxyamphetamine, p hydroxymethamphetamine, amphetamine and norketamine, were simultaneously quantified by the new method (50-5000 ng/ml). The coefficients of variation and the percent deviations for the eight compounds were in the range of 0.2 to 5.3% and -9.4 to +12.8%, respectively. The recoveries were over 90% in all biological samples tested. This method was effective for the separation and the identification of ATSs and their main metabolites having amine moieties in plasma, urine and bile, and was applicable to pharmacokinetic analysis of methamphetamine, ketamine and their main metabolites in biological samples. This analytical method should be useful for the pharmacokinetic analysis of ATSs. PMID- 18396473 TI - Development of porous lamellar poly(L-lactic acid) scaffolds by conventional injection molding process. AB - A novel fabrication technique is proposed for the preparation of unidirectionally oriented, porous scaffolds by selective polymer leaching from lamellar structures created by conventional injection molding. The proof of the concept is implemented using a 50/50 wt.% poly(L-lactic acid)/poly(ethylene oxide) (PLLA/PEO) blend. With this composition, the PLLA and PEO blend is biphasic, containing a homogeneous PLLA/PEO phase and a PEO-rich phase. The two phases were structured using injection molding into well-defined alternating layers of homogeneous PLLA/PEO phase and PEO-rich phase. Leaching of water-soluble PEO from the PEO-rich phase produces macropores, and leaching of phase-separated PEO from the initially homogeneous PLLA/PEO phase produces micropores in the lamellae. Thus, scaffolds with a macroporous lamellar architecture with microporous walls can be produced. The lamellae are continuous along the flow direction, and a continuous lamellar thickness of less than 1 microm could be achieved. Porosities of 57-74% and pore sizes of around 50-100 microm can be obtained using this process. The tensile elastic moduli of the porous constructs were between 580 and 800 MPa. We propose that this organic-solvent-free method of preparing lamellar scaffolds with good mechanical properties, and the reproducibility associated with the injection molding technique, holds promise for a wide range of guided tissue engineering applications. PMID- 18396474 TI - Fetal anemia due to non-Rhesus-D red-cell alloimmunization. AB - Although anti-RhD was once the major etiology of hemolytic disease of the fetus/newborn (HDFN), the widespread adoption of antenatal and postpartum Rhesus immune globulin has resulted in a marked decrease in the prevalence of alloimmunization to the RhD antigen in pregnancy. Maternal alloimmunization to other red cell antigens continues to play a role as the cause of fetal disease since no prophylactic immune globulins are available to prevent the formation of these antibodies. An increasing incidence of the Kell (anti-K1) antibody has been noted in the United States. Guidelines for intervention in cases of irregular red cell antibodies are limited by the bias of anecdotal reports in the literature in favor of severe cases of HFDN. Although most diagnostic protocols used in the management of the RhD-alloimmunized pregnancy can be applied in cases of non-RhD sensitization, Kell (K1 and K2) alloimmunization should be managed more conservatively. PMID- 18396475 TI - Compare and conquer. PMID- 18396476 TI - A double-edged sword. PMID- 18396477 TI - Vaccine partnerships to tackle neglected diseases. PMID- 18396478 TI - AFM-based lithography for nanoscale protein assays. PMID- 18396479 TI - FTIR spectroscopy for grape and wine analysis. PMID- 18396480 TI - Status report on Swedish work environment research--history, context and international evaluation. Introduction. PMID- 18396481 TI - The role of fisheries-induced evolution. PMID- 18396483 TI - The etiology of portal vein thrombosis in patients with hepatocellular carcinoma requires a vigilant diagnostic approach. PMID- 18396482 TI - Characterization of the Swedish work environment and work environment research. PMID- 18396484 TI - Different liver transplant survival rates across the pond. PMID- 18396486 TI - Massive pulmonary embolism during liver transplantation. PMID- 18396485 TI - The classic. Recurrent dislocation of the shoulder joint. Ann Surg. 1950;132:1052 1065. PMID- 18396488 TI - QIOs must show and prove: CMS. PMID- 18396487 TI - A projection pursuit algorithm to classify individuals using fMRI data: Application to schizophrenia. AB - Schizophrenia is diagnosed based largely upon behavioral symptoms. Currently, no quantitative, biologically based diagnostic technique has yet been developed to identify patients with schizophrenia. Classification of individuals into patient with schizophrenia and healthy control groups based on quantitative biologically based data is of great interest to support and refine psychiatric diagnoses. We applied a novel projection pursuit technique on various components obtained with independent component analysis (ICA) of 70 subjects' fMRI activation maps obtained during an auditory oddball task. The validity of the technique was tested with a leave-one-out method and the detection performance varied between 80% and 90%. The findings suggest that the proposed data reduction algorithm is effective in classifying individuals into schizophrenia and healthy control groups and may eventually prove useful as a diagnostic tool. PMID- 18396489 TI - Clinton wins healthcare 'vote'. Democratic primary voters rank issue as a top concern. PMID- 18396490 TI - Easing path for critical access. CMS revisions make it easier for hospitals to rebuild. PMID- 18396491 TI - After the fall. Former HRDI members regroup; some continue to sell expertise to vendors. PMID- 18396492 TI - Trustees of the year 2008. 'An important thing to be part of'. PMID- 18396493 TI - Trustees of the year 2008. There almost every day. PMID- 18396494 TI - Share, and share alike. Proposed rule on PSOs could lead to more data-sharing. PMID- 18396496 TI - Hospitals may join effort. Panel will address racial disparities in healthcare. PMID- 18396495 TI - Primary-care worries. Foreign graduates behind 6% jump: GAO. PMID- 18396497 TI - Prime beef. Calif. chain sues Kaiser over claim denials. PMID- 18396498 TI - Brain drain. As the VA woos behavioral-health providers with bonuses and bigger paychecks, the private sector is feeling the pinch. PMID- 18396499 TI - ROI analysis demonstrates value. PMID- 18396500 TI - [Spondylodiscitis as cause of unexplained fever]. PMID- 18396501 TI - Almost in bloom. PMID- 18396502 TI - Masaaki Ohkoshi Award. PMID- 18396503 TI - Fluoroscopic imaging of epidural intravascular contrast spread in a cadaveric specimen. PMID- 18396504 TI - Bibliography. Current world literature. Nutrition and the intensive care unit. PMID- 18396505 TI - Challenging the three-pore model. PMID- 18396506 TI - Biosurveillance, classification, and semantic health technologies. PMID- 18396507 TI - Augmentation strategies for depression: history and concepts. PMID- 18396508 TI - Augmentation strategies in the treatment of major depressive disorder. Recent findings and current status of augmentation strategies. PMID- 18396509 TI - Augmentation strategies in the treatment of major depressive disorder. Examining the evidence on augmentation with atypical antipsychotics. PMID- 18396510 TI - From childhood into adulthood: the changing face of ADHD. PMID- 18396511 TI - Age-related considerations in the treatment of ADHD. PMID- 18396512 TI - Standing in the shadows of theory. PMID- 18396513 TI - Secondary outputs of alpha1-antitrypsin deficiency targeted detection programme. AB - Targeted detection programmes are recommended to identify subjects affected by severe alpha1-antitrypsin deficiency (AATD). Guidelines are available to address physicians towards subjects at high risk for AATD. We wanted to investigate the clinical characteristics of subjects enrolled in the programme, who result as not being affected by severe AATD; this information is not available in the present literature. We elaborated data contained in the questionnaires accompanying the samples of 2127 Italian subjects submitted for AATD detection in a period spanning 11 years (1996-2006). A total of 588 subjects were eligible to enter this study: PI*MM subjects and subjects with intermediate AATD, referred for lung disease, were characterised by a relatively young mean age, and a high proportion (31.2% and 28.6%, respectively) were never smokers. Fifty percent or more had symptoms of chronic bronchitis, but without obstruction. Only a minority belonged to most severe GOLD stages. The mean levels of AAT varied as a function of the presence or absence of airflow obstruction in intermediate AATD subjects, but not in PI*MM. Individuals enrolled in AATD detection programmes represent an interesting cohort both for public health and research purposes. PMID- 18396514 TI - Proceedings of the International Congress: Stem cells; what future for therapy? Scientific aspects and bioethical problems. PMID- 18396515 TI - Proceedings of the IVth Franco-Australian Meeting on Hypertension, 9-12 September 2007, Australia. PMID- 18396516 TI - Endothelin-1 in exhaled breath condensate of stable and unstable asthma patients. AB - Endothelins are proinflammatory, profibrotic, broncho- and vasoconstrictive peptides, which play an important role in the development of airway inflammation and remodeling in asthma. The study was undertaken to evaluate the endothelin-1 (ET-1) levels in exhaled breath condensate (EBC) of asthmatics with different degree in asthma severity. EBC was collected from 31 patients with allergic asthma (11 with steroid-naive mild asthma, 10 with ICS-treated, stable mild-to moderate asthma, 10 with ICS-treated unstable, severe asthma) and 7 healthy volunteers. In the three groups of asthmatics, ET-1 concentrations in EBC were significantly higher than in healthy volunteers. ET-1 levels were significantly higher in patients with unstable asthma than in the two groups with stable disease. There was a significant correlation between ET-1 levels and FENO in the three groups of asthmatics and between ET-1 and blood eosinophil counts in the group of patients with unstable asthma. Measurements of ET-1 in EBC may provide another useful diagnostic tool for detecting and monitoring inflammation in patients with asthma. PMID- 18396517 TI - [ Standards, Options and Recommendations for the management of patients with epithelial ovarian cancer--surgical treatment update 2007 (summary report)]. PMID- 18396518 TI - What lies between hypomania and bipolar disorder? A qualitative analysis of 12 non-treatment-seeking people with a history of hypomanic experiences and no history of major depression. AB - This study presents the thoughts of a group of 12 individuals over 30 years of age with a history of hypomanic experiences using the qualitative method: Interpretative Phenomenological Analysis. These individuals were specifically selected on the basis of a self-reported history of hypomanic experiences (according to the mood disorder questionnaire, MDQ), and screened to rule out a history of mania or major depression, and therefore not fulfilling the criteria for bipolar I or II disorders. Thus, the current participants represent a unique sample of individuals, previously underreported in the literature. The aim of the interview was to determine the protective factors or characteristics which may prevent such individuals from experiencing hypomanic experiences that significantly disrupt their level of functioning. Three clusters of emergent themes emerged in the interviews: 'positive qualities of hypomanic experiences', 'the social meaning of hypomanic experiences', and 'having hypomanic experiences is not a problem'. The responses to several interview questions are also provided, which highlight the situations preceding hypomanic experiences, possible explanations for their experiences not going out of control, and typical responses of other people to the participants' behaviour. Findings are discussed in relation to a recent cognitive model of bipolar disorder and mood swings. PMID- 18396519 TI - Building up to the Darwin double. PMID- 18396520 TI - Defibrillators' weak link. PMID- 18396521 TI - [Evaluating general and occupational morbidity on mining enterprises of Kazakhstan]. AB - Evaluating risks of occupational and occupationally mediated diseases is a main parameter in further social and hygienic monitoring of work conditions and health state among mining industry workers. PMID- 18396522 TI - [Hygienic, ecologic and medical problems in area influenced by space rockets launching]. AB - The article deals with characteristics and features of morbidity structure among population in area influenced by space rockets launching. PMID- 18396523 TI - [Contemporary state of using specialized food products to correct metabolic processes in chemical intoxication (review of literature)]. AB - The article deals with topics associated with contemporary state of using specialized food products to correct metabolic processes in intoxication with various chemicals. PMID- 18396524 TI - Leadership lacking. PMID- 18396525 TI - The joys of patient care. PMID- 18396526 TI - What is happening to the profession? PMID- 18396527 TI - Violence increases yet staff still await training. PMID- 18396528 TI - Payment by results must 'do what it says on the tin'. PMID- 18396529 TI - Cuts stifle talking therapy. PMID- 18396530 TI - Call to address 'cradle to grave gap'. PMID- 18396531 TI - Welsh epilepsy plan first of its kind. PMID- 18396532 TI - Innovation under wraps. AB - New technologies in the NHS tend to create higher demand. The commons health select committee recently recommended that NICE extends its cost-evaluating remit. In the meantime some believe PCTs should work together on decision making about costly new interventions. PMID- 18396533 TI - What a picture. PMID- 18396534 TI - Clinical engagement. Surrender to progress. PMID- 18396535 TI - [Instruments, books and other objects memorable to (almost) forgotten opinions, therapies, buildings, et cetera. Bull guide]. PMID- 18396537 TI - Simultaneous chemical and sensory characterization of volatile organic compounds and semi-volatile organic compounds emitted from swine manure using solid phase microextraction and multidimensional gas chromatography-mass spectrometry olfactometry. AB - Swine manure is associated with emissions of odor, volatile organic compounds (VOCs) and other gases that can affect air quality on local and regional scales. In this research, a solid phase microextraction (SPME) and novel multidimensional gas chromatography-mass spectrometry-olfactometry (MDGC-MS-O) system were used to simultaneously identify VOCs and related odors emitted from swine manure. Gas samples were extracted from manure headspace using Carboxen/polydimethylsiloxane (PDMS) 85-microm SPME fibers. The MDGC-MS-O system was equipped with two columns in series with a system of valves allowing transfer of samples between columns (heartcutting). The heartcuts were used to maximize the isolation, separation, and identification of compounds. The odor impact of separated compounds was evaluated by a trained panelist for character and intensity. A total of 295 compounds with molecular weights ranging from 34 to 260 were identified. Seventy one compounds had a distinct odor. Nearly 68% of the compounds for which reaction rates with OH* radicals are known had an estimated atmospheric lifetime <24 h. PMID- 18396538 TI - Carbon dioxide efflux from soil with poultry litter applications in conventional and conservation tillage systems in northern Alabama. AB - Increased CO2 release from soils resulting from agricultural practices such as tillage has generated concerns about contributions to global warming. Maintaining current levels of soil C and/or sequestering additional C in soils are important mechanisms to reduce CO2 in the atmosphere through production agriculture. We conducted a study in northern Alabama from 2003 to 2006 to measure CO2 efflux and C storage in long-term tilled and non-tilled cotton (Gossypium hirsutum L.) plots receiving poultry litter or ammonium nitrate (AN). Treatments were established in 1996 on a Decatur silt loam (clayey, kaolinitic thermic, Typic Paleudults) and consisted of conventional-tillage (CT), mulch-tillage (MT), and no-tillage (NT) systems with winter rye [Secale cereale (L.)] cover cropping and AN and poultry litter (PL) as nitrogen sources. Cotton was planted in 2003, 2004, and 2006. Corn was planted in 2005 as a rotation crop using a no-till planter in all plots, and no fertilizer was applied. Poultry litter application resulted in higher CO2 emission from soil compared with AN application regardless of tillage system. In 2003 and 2006, CT (4.39 and 3.40 micromol m(-2) s(-1), respectively) and MT (4.17 and 3.39 micromol m(-2) s(-1), respectively) with PL at 100 kg N ha(-1) (100 PLN) recorded significantly higher CO2 efflux compared with NT with 100 PLN (2.84 and 2.47 micromol m(-2) s(-1), respectively). Total soil C at 0- to 15-cm depth was not affected by tillage but significantly increased with PL application and winter rye cover cropping. In general, cotton produced with NT conservation tillage in conjunction with PL and winter rye cover cropping reduced CO2 emissions and sequestered more soil C compared with control treatments. PMID- 18396539 TI - [Evaluation of sperm mitochondrial membrane potential by JC-1 fluorescent staining and flow cytometry]. AB - OBJECTIVE: To investigate the feasibility and clinical significance of detecting sperm mitochondrial membrane potential (MMP) by JC-1 fluorescent staining and flow cytometry, and to explore the relationship between the results of JC-1 staining and seminal parameters. METHODS: Sixty-three semen samples were divided into a fertile (n = 31) and an infertile group (n = 32) and underwent computer assisted semen analysis (CASA). All the samples were washed, followed by JC-1 staining and evaluation of sperm MMP by flow cytometry. The percentage of normal sperm MMP was indicated as the percentage of sperm emitting orange-red fluorescence (JC-1 + %). RESULTS: The JC-1 + % was significantly higher in the fertile group than in the infertile one ([75.89 +/- 15.69]% vs [54.04 +/- 22.21] %, P = 0.000), correlated positively with sperm motility (r = 0.610, P = 0.000) and the percentage of grade a + b sperm (r = 0.614, P = 0.000) and negatively with grade d sperm (r = -0.504, P = 0.000). There was a significant positive correlation between the results of JC-1 staining (JC-1 + %) and that of Rh123 /PI dual fluorescent staining (Rh123 + / PI (-)%) (r = 0.938, P = 0.000). CONCLUSION: JC-1 staining and flow cytometry could readily and quickly detect sperm MMP and the sperm JC-1 + % could be an auxiliary marker for the diagnosis of male infertility. PMID- 18396540 TI - Near-field dust exposure from cotton field tilling and harvesting. AB - The frequency and intensities of dust exposures in and near farm fields, which potentially contribute to high intensity human exposure events, are undocumented due to the transient nature of local dust plumes and the difficulties of making accurate concentration measurements. The objective of this study is to measure near-field spatial concentrations of the dust plumes emitted during tilling and harvesting of an irrigated cotton field outside of Las Cruces, NM (soil class: fine-loamy, mixed, superactive, thermic Typic Calciargid). A comparison of remote lidar measurements of plumes emitted from cotton field operations with in situ samplers shows a strong agreement between the two techniques: r2 = 0.79 for total suspended particulates (TSP) and r2 = 0.61 for particulate matter with diameter less than or equal to 10 microm (PM10). Plume movement was dependent on the short term wind field and atmospheric stability. Horizontal spread rate of the plumes, determined from lidar measured Gaussian dispersion parameters, was less than expected by a factor of 7. Thus, in-plume downwind concentrations were higher than expected. Vertical dispersion was dependent on the rise of "cells" of warm air convecting off the soil surface. On a windy day, discing the field showed TSP and PM10 concentrations at the source itself of up to 176 microg m(-3) and 120 microg m(-3), respectively. These resulted in in-plume peak TSP concentrations of about 1.22 microg m(-3) at 10 m downwind and 0.33 microg m(-3) at 100 m downwind. The measured concentrations highlight a potential exposure risk to people in and around farming operations. PMID- 18396541 TI - Development of a dynamic model to predict PM10 emissions from swine houses. AB - Influences on dust emissions from livestock operations are number, weight, and kind of animals and characteristics of the housing system. Differences between facilities cannot be explained solely by mechanistic input variables. The objective of this study was to characterize the main input variables for modeling emissions of particulate matter with a mass median diameter < or = 10 microm (PM10) from swine facilities using a data-based model. Investigations were performed in mechanically ventilated facilities for weaning, growing-finishing, and sows in Italy and Germany. The measurements included inside and outside concentration of airborne PM10 particles (scatter light photometry), ventilation rate (calibrated measuring fans), indoor air climate at a measuring frequency of 60 s, feeding times, and animal-related data such as weight and animal activity. Dust concentration and emission were simulated using a dynamic transfer function. The results indicated that the average PM10 emission rate was influenced considerably by housing system. The simulation of the PM10 emission rate resulted in a mean percentage error per data set of 21 to 39%, whereas the average simulated and measured emission rate per data set differed by about 4 to 19%. High prediction errors occurred especially during situations in which the absolute level and spatial location of the measured activity peaks did not correspond with the measured dust peaks. Further recommendations of the study were to improve continuous and accurate measurements of input variables, such as the activity level in animal houses, and to optimize the amount of measuring days in relation to the model accuracy. PMID- 18396542 TI - Abundances and flux estimates of volatile organic compounds from a dairy cowshed in Germany. AB - Animal husbandry and manure treatment have been specifically documented as significant sources of methane, ammonia, nitrous oxide, and particulate matter. Although volatile organic compounds (VOCs) are also produced, much less information exists concerning their impact. We report on chemical ionization mass spectrometry and photo-acoustic spectroscopy measurements of mixing ratios of VOCs over a 2-wk measurement period in a large cowshed at the Federal Agricultural Research Centre (FAL) in Mariensee, Germany. The high time resolution of these measurements enables insight into the sources of the emissions in a typical livestock management setting. During feeding hours and solid manure removal, large mixing ratio spikes of several VOCs were observed and correlated with simultaneous methane, carbon dioxide, and ammonia level enhancements. The subsequent decay of cowshed concentration due to passive cowshed ventilation was used to model emission rates, which were dominated by ethanol and acetic acid, followed by methanol. Correlations of VOC mixing ratios with methane or ammonia were also used to calculate cowshed emission factors and to estimate potential nationwide VOC emissions from dairy cows. The results ranged from around 0.1 Gg carbon per year (1 Gg = 10(9) g) for nonanal and dimethylsulfide, several Gg carbon per year for volatile fatty acids and methanol, to over 10 Gg carbon per year of emitted ethanol. While some estimates were not consistent between the two extrapolation methods, the results indicate that animal husbandry VOC emissions are dominated by oxygenated compounds and may be a nationally but not globally significant emission to the atmosphere. PMID- 18396543 TI - In situ transmissiometer measurements for real-time monitoring of dust discharge during orchard nut harvesting. AB - Rapid assessments of operating conditions and field preparation on dust discharge from nut harvesters are needed to guide improved equipment design and grower practices for dust reduction. An industrial opacity sensor, typically used for industrial stack monitoring, was adapted for use on a nut harvester to measure relative dust intensity during nut pick-up operations in almond orchards. Due to the high volume of discharge air and the presence of large debris such as leaves, additional components were coupled with the sensor to enable subsampling of the air. Pre-harvest windrow preparation conditions were evaluated. Results indicated that relative dust intensity decreased by 32% during harvest activities after windrow preparation with proper nut sweeper adjustment. Conventional harvesting results indicated that under typical operating conditions, reducing the separation fan speed could reduce relative dust intensity by 54%. Ground speed also had a strong effect; reducing speed from 4.8 to 2.4 km h(-1) reduced opacity of discharged air by 50%. The measurement system was also mounted on a separate vehicle and used as a tool for comparing modifications in harvest machine designs where direct measurement of discharge may not be feasible due to mechanical constraints. A comparison between a conventional harvester and one modification in the harvester design found that the machine modification decreased relative dust intensity by 73%. The measurement tools described in this work can be used to provide rapid feedback on harvester operating conditions, orchard cultural practices, and machine design modifications. PMID- 18396544 TI - Methane emissions from free-ranging cattle: comparison of tracer and integrated horizontal flux techniques. AB - Accurate measurements of methane (CH4) emission rates from livestock in their undisturbed natural environments are required to assess their impacts on radiative forcing (i.e., enhanced greenhouse effect) and the environment. Here we compare results from two nonintrusive techniques for the measurement of CH4 emissions from cattle. The cows were kept in an outdoor feeding strip that allowed them to follow natural behavioral patterns but contained them within a well defined space. In the first technique, nitrous oxide (N2O) was released as a tracer at the upwind edge of the feeding strip, and the downwind concentrations of N2O and CH4 were measured simultaneously using Fourier transform infrared (FTIR) spectroscopy. Average CH4 emission per cow was calculated each half-hour on three separate days from the correlation between the two gases. The second technique was the integrated horizontal flux (IHF) or 1-D mass-balance method, in which we used the measured vertical profiles of CH4 concentration and windspeed downwind of the cows to determine the total CH4 emission. Comparing the IHF results to the known release rate of N2O allowed us to test the IHF technique independently. We found agreement within 10% for all comparisons on all days. The daily CH4 emission rate averaged over all tracer and IHF measurements was 342 g CH4 head(-1) d(-1). This is within the range of previous measurements for mature lactating dairy cattle (200-430 g CH4 head(-1) d(-1)) but higher than expected for yearling cattle. The high CH4 emissions are accompanied by high CO2 emissions determined from the FTIR measurements. The bias is most likely due to the measurements being made during and after supplementary feeding of the cattle. PMID- 18396545 TI - Carbon monoxide from composting due to thermal oxidation of biomass. AB - Emissions of carbon monoxide (CO) were observed from decomposing organic wastes and litter under laboratory, pilot composting plant, and natural conditions. Field studies included air from inside a compost heap of about 200 m3, emissions from composting of livestock wastes at a biologically operating farm, and leaf litter pile air samples. The concentration of CO was up to 120 micromol mol(-1) in the compost piles of green waste, and up to 10 micromol mol(-1) in flux chambers above livestock waste windrow composts. The mean CO flux rates were approximately 20 mg CO m(-2) h(-1) for compost heaps of green waste, and varied from 30 to 100 mg CO m(-2) h(-1) for fresh dung windrows. Laboratory studies using a temperature and ventilation-controlled substrate container were performed to elucidate the origin of CO, and included hay samples of fixed moisture content at temperatures between 5 and 65 degrees C, including nonsterilized as well as sterilized samples. The concentration of CO was up to 160 micromol mol(-1) in these experiments, and Arrhenius-type plot analyses resulted in activation energies of 65 kJ mol(-1) for thermochemically produced CO from the nonsterilized compost substrate. Sterilized samples showed dramatically reduced CO2 but virtually unchanged CO emissions, albeit at a slightly lower activation energy, likely a result of the high-temperature sterilization. Though globally and regionally these CO emissions are only a minor source, thermochemically produced CO emissions might affect local air quality in and near composting facilities. PMID- 18396546 TI - Soil-atmosphere trace gas exchange in semiarid and arid zones. AB - A review is presented on trace gas exchange of CH4, CO, N2O, and NOx arising from agriculture and natural sources in the world's semiarid and arid zones due to soil processes. These gases are important contributors to the radiative forcing and the chemistry of the atmosphere. Quantitative information is summarized from the available studies. Between 5 and 40% of the global soil-atmosphere exchange for these gases (CH4, CO, N2O, and NOx) may occur in semiarid and arid zones, but for each of these gases there are fewer than a dozen studies to support the individual estimates, and these are from a limited number of locations. Significant differences in the biophysical and chemical processes controlling these trace gas exchanges are identified through the comparison of semiarid and arid zones with the moist temperate or wet/dry savanna land regions. Therefore, there is a poorly quantified understanding of the contribution of these regions to the global trace gas cycles and atmospheric chemistry. More importantly, there is a poor understanding of the feedback between these exchanges, global change, and regional land use and air pollution issues. A set of research issues is presented. PMID- 18396547 TI - Effects of sodium bisulfate on alcohol, amine, and ammonia emissions from dairy slurry. AB - Sodium bisulfate (SBS) is extensively used in the poultry industry to reduce ammonia and bacterial levels in litter. It is also used in the dairy industry to reduce bacterial counts in bedding and ammonia emissions, preventing environmental mastitis and calf respiratory stress. The present study measured the effect of SBS on the air emission of ammonia, amine, and alcohol from a dairy slurry mix. Amine flux was undetectable (<5 ng L(-1)) across treatments. Application of SBS decreased ammonia, methanol, and ethanol emissions from fresh dairy slurry. Ammonia emissions decreased with increasing levels of SBS treatment. The 3-d average ammonia flux from the control (no SBS applied) and the three different SBS surface application levels of 0.125, 0.250, and 0.375 kg m( 2) were 513.4, 407.2, 294.8, and 204.5 mg h(-1) m(-2), respectively. The ammonia emission reduction potentials were 0, 21, 43, and 60%, respectively. Methanol and ethanol emissions decreased with an increase in the amount of SBS applied. The 3 d average methanol emissions were 223.7, 178.0, 131.6, and 87.0 mg h(-1) m(-2) for SBS surface application level of 0, 0.125, 0.250, and 0.375 kg m(-2), with corresponding reduction potentials of 0, 20, 41, and 61, respectively. Similar emission reduction potentials of 0, 18, 35, and 58% were obtained for ethanol. Sodium bisulfate was shown to be effective in the mitigation of ammonia and alcohol emissions from fresh dairy slurry. PMID- 18396548 TI - Alcohol, volatile fatty acid, phenol, and methane emissions from dairy cows and fresh manure. AB - There are approximately 2.5 million dairy cows in California. Emission inventories list dairy cows and their manure as the major source of regional air pollutants, but data on their actual emissions remain sparse, particularly for smog-forming volatile organic compounds (VOCs) and greenhouse gases (GHGs). We report measurements of alcohols, volatile fatty acids, phenols, and methane (CH4) emitted from nonlactating (dry) and lactating dairy cows and their manure under controlled conditions. The experiment was conducted in an environmental chamber that simulates commercial concrete-floored freestall cow housing conditions. The fluxes of methanol, ethanol, and CH4 were measured from cows and/or their fresh manure. The average estimated methanol and ethanol emissions were 0.33 and 0.51 g cow(-1) h(-1) from dry cows and manure and 0.7 and 1.27 g cow(-1) h(-1) from lactating cows and manure, respectively. Both alcohols increased over time, coinciding with increasing accumulation of manure on the chamber floor. Volatile fatty acids and phenols were emitted at concentrations close to their detection limit. Average estimated CH4 emissions were predominantly associated with enteric fermentation from cows rather than manure and were 12.35 and 18.23 g cow(-1) h( 1) for dry and lactating cows, respectively. Lactating cows produced considerably more gaseous VOCs and GHGs emissions than dry cows (P < 0.001). Dairy cows and fresh manure have the potential to emit considerable amounts of alcohols and CH4 and research is needed to determine effective mitigation. PMID- 18396549 TI - Soil quality assessment in rice production systems: establishing a minimum data set. AB - Soil quality, as a measure of the soil's capacity to function, can be assessed by indicators based on physical, chemical, and biological properties. Here we report on the assessment of soil quality in 21 rice (Oryza sativa) fields under three rice production systems (semi-direct, pre-germinated, and conventional) on four soil textural classes in the Camaqua region of Rio Grande do Sul, Brazil. The objectives of our study were: (i) to identify soil quality indicators that discriminate both management systems and soil textural classes, (ii) to establish a minimum data set of soil quality indicators and (iii) to test whether this minimum data set is correlated with yield. Twenty-nine soil biological, chemical, and physical properties were evaluated to characterize regional soil quality. Soil quality assessment was based on factor and discriminant analysis. Bulk density, available water, and micronutrients (Cu, Zn, and Mn) were the most powerful soil properties in distinguishing among different soil textural classes. Organic matter, earthworms, micronutrients (Cu and Mn), and mean weight diameter were the most powerful soil properties in assessing differences in soil quality among the rice management systems. Manganese was the property most strongly correlated with yield (adjusted r2 = 0.365, P = 0.001). The merits of sub dividing samples according to texture and the linkage between soil quality indicators, soil functioning, plant performance, and soil management options are discussed in particular. PMID- 18396550 TI - Use of a low-cost biosorbent to remove pesticides from wastewater. AB - A lignocellulosic substrate (LS) obtained from our local agroindustry was used as a low-cost and effective adsorbent for the removal of pesticides from wastewaters. The studied pesticides were terbumeton (N-(1,1-dimethyl)-Nethyl-6 methoxy-1,3,5-triazine-2,4-diamine), desethyl terbumeton (N-(1,1-dimethylethyl)-6 methoxy-1,3,5-triazine-2,4-diamine), dimetomorph (4-[3-(4-chlorophenyl)-3-(3,4 dimethoxyphenyl)acryloyl]morpholine), and isoproturon (3-(4-isopropylphenyl)-1,1 dimethylurea). Batch and column experiments were conducted as a function of pH and pesticide concentration under laboratory and industrial conditions. The concentration range studied for the pesticides varied from 2 x 10(-7) to 3 x 10( 4) mol L(-1). The influence of organic and inorganic pollutants was assessed by studying the retention of pesticide in the presence of copper(II) and a surfactant. These experiments indicated that LS is an efficient adsorbent toward the investigated pesticides and has little influence of the other pollutants. The kinetic adsorptions are fast, and the amounts of adsorbed pesticide varied from 1 to 8 g kg(-1) of LS. These retention capacities show that LS can provide a simple, effective, and cheap method for removing pesticides from contaminated waters. Thus, this biomaterial may be useful for cleaning up polluted waters. PMID- 18396551 TI - Are leaves that fall from imidacloprid-treated maple trees to control Asian longhorned beetles toxic to non-target decomposer organisms? AB - The systemic insecticide imidacloprid may be applied to deciduous trees for control of the Asian longhorned beetle, an invasive wood-boring insect. Senescent leaves falling from systemically treated trees contain imidacloprid concentrations that could pose a risk to natural decomposer organisms. We examined the effects of foliar imidacloprid concentrations on decomposer organisms by adding leaves from imidacloprid-treated sugar maple trees to aquatic and terrestrial microcosms under controlled laboratory conditions. Imidacloprid in maple leaves at realistic field concentrations (3-11 mg kg(-1)) did not affect survival of aquatic leaf-shredding insects or litter-dwelling earthworms. However, adverse sublethal effects at these concentrations were detected. Feeding rates by aquatic insects and earthworms were reduced, leaf decomposition (mass loss) was decreased, measurable weight losses occurred among earthworms, and aquatic and terrestrial microbial decomposition activity was significantly inhibited. Results of this study suggest that sugar maple trees systemically treated with imidacloprid to control Asian longhorned beetles may yield senescent leaves with residue levels sufficient to reduce natural decomposition processes in aquatic and terrestrial environments through adverse effects on non-target decomposer organisms. PMID- 18396552 TI - Microbial populations and enzyme activities in soil in situ under transgenic corn expressing cry proteins from Bacillus thuringiensis. AB - Transgenic Bt crops produce insecticidal Cry proteins that are released to soil in plant residues, root exudates, and pollen and that may affect soil microorganisms. As a continuation of studies in the laboratory and a plant-growth room, a field study was conducted at the Rosemount Experiment Station of the University of Minnesota. Three Bt corn varieties that express the Cry1Ab protein, which is toxic to the European corn borer (Ostrinia nubilalis Hubner), and one Bt corn variety that expresses the Cry3Bb1 protein, which is toxic to the corn rootworm complex (Diabrotica spp.), and their near-isogenic non-Bt varieties were evaluated for their effects on microbial diversity by classical dilution plating and molecular (polymerase chain reaction-denaturing gradient gel electrophoresis) techniques and for the activities of some enzymes (arylsulfatases, acid and alkaline phosphatases, dehydrogenases, and proteases) involved in the degradation of plant biomass. After 4 consecutive years of corn cultivation (2003-2006), there were, in general, no consistent statistically significant differences in the numbers of different groups of microorganisms, the activities of the enzymes, and the pH between soils planted with Bt and non-Bt corn. Numbers and types of microorganisms and enzyme activities differed with season and with the varieties of corn, but these differences were not related to the presence of the Cry proteins in soil. The Cry1Ab protein of Bt corn (events Bt11 and MON810) was detected in most soils during the 4 yr, whereas the Cry3Bb1 protein was not detected in soils of Bt corn (event MON863) expressing the cry3Bb1 gene. PMID- 18396553 TI - Storage and dynamics of carbon and nitrogen in soil after grazing exclusion in Leymus chinensis grasslands of northern China. AB - Land-use change can lead to changes in soil carbon (C) and nitrogen (N) storage. This study aimed to determine the impact of long-term grazing exclusion (GE) on soil organic C and total N (TN) storage in the Leymus chinensis grasslands of northern China and to estimate the dynamics of recovery after GE. We investigated the aboveground biomass and soil organic C and TN storage in six contiguous plots along a GE chronosequence comprising free grazing, 3-yr GE, 8-yr GE, 20-yr GE, 24 yr GE, and 28-yr GE. Grazing exclusion for two decades increased the soil C and N storage by 35.7 and 14.6%, respectively, in the 0- to 40-cm soil layer. The aboveground net primary productivity and soil C and N storage were the highest with 24-yr GE and the lowest with free grazing. The storage increased logarithmically with the duration of GE; after an initial rapid increase after the introduction of GE, the storage attained equilibrium after 20 yr. A logarithmic regression analysis revealed 86.8 and 87.1% variation in the soil C storage and 74.2 and 80.7% variation in the soil N storage in the 0- to 10-cm and 0- to 40-cm soil layers, respectively. Based on these results, we suggest that two decades of GE would restore the L. chinensis grasslands from being lightly degraded to a stable productive condition with good soil C and N storage capacity. Our results demonstrated that by implementing GE, the temperate grasslands of northern China could facilitate significant C and N storage on decade scales in the context of mitigating global climate change. PMID- 18396554 TI - Persistent elevated nitrate in a riparian zone aquifer. AB - Streamside vegetated buffer strips (riparian zones) are often assumed to be zones of ground water nitrate (NO3(-)) attenuation. At a site in southwestern Ontario (Zorra site), detailed monitoring revealed that elevated NO3(-) -N (4-93 mg L( 1)) persisted throughout a 100-m-wide riparian floodplain. Typical of riparian zones, the site has a soil zone of recent river alluvium that is organic carbon (OC) rich (36 +/- 16 g kg(-1)). This material is underlain by an older glacial outwash aquifer with a much lower OC content (2.3 +/- 2.5 g kg(-1). Examination of NO3(-), Cl(-), SO4(2-), and dissolved organic carbon (DOC) concentrations; N/Cl ratios; and NO3(-) isotopic composition (delta15N and delta18O) provides evidence of four distinct NO3(-) source zones within the riparian environment. Denitrification occurs but is incomplete and is restricted to a narrow interval located within ~0.5 m of the alluvium-aquifer contact and to one zone (poultry manure compost zone) where elevated DOC persists from the source. In older ground water close to the river discharge point, denitrification remains insufficient to substantially deplete NO3(-). Overall, denitrification related specifically to the riparian environment is limited at this site. The persistence of NO3(-) in the aquifer at this site is a consequence of its Pleistocene age and resulting low OC content, in contrast to recent fluvial sediments in modern agricultural terrain, which, even if permeable, usually have zones enriched in labile OC. Thus, sediment age and origin are additional factors that should be considered when assessing the potential for riparian zone denitrification. PMID- 18396555 TI - Adsorption-desorption characteristics of mercury in paddy soils of China. AB - Mercury (Hg) has received considerable attention because of its association with various human health problems. Adsorption-desorption behavior of Hg at contaminated levels in two paddy soils was investigated. The two representative soils for rice production in China, locally referred to as a yellowish red soil (YRS) and silty loam soil (SLS) and classified as Gleyi-Stagnic Anthrosols in FAO/UNESCO nomenclature, were respectively collected from Jiaxin County and Xiasha District of Hangzhou City, Zhejiang Province. The YRS adsorbed more Hg(2+) than the SLS. The characteristics of Hg adsorption could be described by the simple Langmuir adsorption equation (r2 = 0.999 and 0.999, P < 0.01, respectively, for the SLS and YRS). The maximum adsorption values (Xm) that were obtained from the simple Langmuir model were 111 and 213 mg Hg(2+) kg(-1) soil, respectively, for the SLS and YRS. Adsorption of Hg(2+) decreased soil pH by 0.75 unit for the SLS soil and 0.91 unit for the YRS soil at the highest loading. The distribution coefficient (kd) of Hg in the soil decreased exponentially with increasing Hg(2+) loading. After five successive desorptions with 0.01 mol L(-1) KCl solution (pH 5.4), 0 to 24.4% of the total adsorbed Hg(2+) in the SLS soil was desorbed and the corresponding value of the YRS soil was 0 to 14.4%, indicating that the SLS soil had a lower affinity for Hg(2+) than the YRS soil at the same Hg(2+) loading. Different mechanisms are likely involved in Hg(2+) adsorption-desorption at different levels of Hg(2+) loading and between the two soils. PMID- 18396556 TI - Arsenic, cadmium, and lead in California cropland soils: role of phosphate and micronutrient fertilizers. AB - Phosphate and micronutrient fertilizers contain potentially harmful trace elements, such as arsenic (As), cadmium (Cd), and lead (Pb). We investigated if application of these fertilizer increases the As, Cd, and Pb concentrations of the receiving soils. More than 1000 soil samples were collected in seven major vegetable production regions across California. Benchmark soils (no or low fertilizer input) sampled in 1967 and re-sampled in 2001 served as a baseline. Soils were analyzed for total concentrations of As, Cd, Pb, P, and Zn. The P and Zn concentrations of the soils were indicators of P fertilizer and micronutrient inputs, respectively. Results showed that the concentrations of these elements in the vegetable production fields in some production areas of California had been shifted upward. Principal component analysis and cluster analysis showed that the seven production areas could be sorted into three categories: (i) enrichment of As, Cd, and Pb, which was associated with the enrichment of P and Zn in one of the seven areas surveyed; (ii) enrichment of As, which was associated with enrichment of Zn in two of the seven areas surveyed; and (iii) no remarkable correlation between enrichment of As, Cd, and Pb and enrichment of P and Zn in the other four areas surveyed. PMID- 18396557 TI - The impact of agricultural runoff on the quality of two streams in vegetable farm areas in Ghana. AB - A study of two small streams at Akumadan and Tono, Ghana, was undertaken during the rain and dry season periods between February 2005 and January 2006 to investigate the impact of vegetable field runoff on their quality. In each stream we compared the concentration of current-use pesticides in one site immediately upstream of a vegetable field with a second site immediately downstream. Only trace concentrations of endosulfan and chlorpyrifos were detected at both sites in both streams in the dry season. In the wet season, rain-induced runoff transported pesticides into downstream stretches of the streams. Average peak levels in the streams themselves were 0.07 microg L(-1) endosulfan, 0.02 microg L(-1) chlorpyrifos (the Akumadan stream); 0.04 microg L(-1) endosulfan, 0.02 microg L(-1) chlorpyrifos (the Tono stream). Respective average pesticide levels associated with streambed sediment were 1.34 and 0.32 microg kg(-1) (the Akumadan stream), and 0.92 and 0.84 microg kg(-1) (the Tono stream). Further investigations are needed to establish the potential endosulfan and chlorpyrifos effects on aquatic invertebrate and fish in these streams. Meanwhile measures should be undertaken to reduce the input of these chemicals via runoff. PMID- 18396558 TI - Sorption of the antibiotic tetracycline to humic-mineral complexes. AB - Humic substances, by altering the surface properties and/or competing for available reaction sites, can either suppress or promote sorption of organic compounds to mineral surfaces. Limited literature evidence points to the reduction in sorption of the antibiotic tetracycline to clay minerals in the presence of humic substances. We investigated the surface interaction of Elliott soil humic acid (ESHA) with hydrous Al oxide (HAO) and the effect of this association on tetracycline sorption. Strong interaction between ESHA and HAO led to ESHA-promoted dissolution of HAO and surface charge reversal. The ESHA-HAO sorption-desorption isotherms were successfully described using a modified Langmuir model that accounted for the heterogeneity of HAO surface and ESHA. Ligand exchange was proposed as the major interaction mechanism, and the edge Al atoms on HAO surface were considered as the sorption sites for ESHA macromolecules. ESHA was coated onto HAO to achieve two different organic content (foc) levels of 0.81 and 1.52%. Sorption results were compared for the binary ESHA-tetracycline and HAO-tetracycline systems, and the ternary ESHA-HAO tetracycline system. The coating of ESHA on HAO significantly suppressed tetracycline sorption levels, attributable to altered HAO surface charge characteristics and/or direct competition between ESHA and tetracycline for potential sorption sites. Higher foc level, besides increasing the extent of sorption suppression, also resulted in greater ionic strength dependence and increased nonlinearity of sorption behavior. It, therefore, appears that the presence of humic substances, in both dissolved and mineral-bound forms, is likely to increase the environmental mobility of tetracycline compounds. PMID- 18396559 TI - Nitrogen loss through lateral seepage in near-trench paddy fields. AB - A near-trench paddy field experiment with five urea application rates (0-360 kg N ha(-1) in 90-kg increments) was conducted on a paddy soil in the Taihu Lake Region of China to elucidate N losses through lateral seepage during three rice (Oryza sativa L.) growing seasons. The total N (Nt), NH4(+) -N, and NO3(-) -N concentrations in the lateral seepage water increased with increasing N rates. The seasonal Nt fluxes by lateral seepage varied from 6.8 to 25.6 kg N ha(-1) for urea application rates of 90 to 360 kg N ha(-1). Lateral seepage accounted for 4.7 to 6.6% of the Nt applied, implying that lateral seepage was an important pathway of N loss from near-trench paddy fields. The cumulative N loss via lateral seepage was significantly related to N fertilization rate (P = 0.05). Floodwater level was also identified as a main factor affecting N losses via lateral seepage from paddy fields, as indicated by a positive linear relationship (R2 = 0.43) between floodwater level and daily lateral flow during the flooded period (P = 0.05). Under the conditions of these experiments, a shallow floodwater depth of 50 mm, urea application rates of 90 kg N ha(-1) or less, and no rainfall within 1 wk after N application reduced N losses by lateral seepage from paddy fields. PMID- 18396560 TI - Managing broiler litter application rate and grazing to decrease watershed runoff losses. AB - Pasture management and broiler litter application rate are critical factors influencing the magnitude of nutrients being transported by runoff from fields. We investigated the impact of pasture management and broiler litter application rate on nutrient runoff from bermudagrass (Cynodon dactylon) pastures. The experiment was conducted on a Ruston fine sandy loam with a factorial arrangement on 21 large paddocks. Runoff water was collected from natural rainfall events from 2001 to 2003. Runoff water and soil samples were analyzed for nutrients and sediments. Runoff was generally greater (29%) from grazed than hayed pastures regardless of the litter application rate. There was greater inorganic N in the runoff from grazed paddocks when litter rate was based on N rather than P. The mean total P loss per runoff event for all treatments ranged from 7 to 45 g ha( 1) and the grazed treatment with litter applied on N basis had the greatest total P loss. Total dissolved P was the dominant P fraction in the runoff, ranging from 85% to 93% of the total P. The soluble reactive P was greater for treatments with litter applied on N basis regardless of pasture management. Runoff total sediments were greater for N-based litter application compared to those which received litter on P basis. Our results indicate that litter may be applied on N basis if the pasture is hayed and the soil P is low. In contrast, litter rates should be based on a P-basis if pasture is grazed. PMID- 18396561 TI - Physical and chemical changes during composting of wood chip-bedded and straw bedded beef cattle feedlot manure. AB - In the 1990s, restrictions on incineration encouraged the forest industry in western Canada to develop new uses for their wood residuals by product. One such use was as a replacement for cereal straw bedding in southern Alberta's beef cattle (Bos taurus) feedlot industry. However, use of carbon (C)-rich bedding, such as wood chips, had implications for subsequent composting of the feedlot manure, a practice that was being increasingly adopted. In a 3-yr study, we compared composting of wood chip-bedded manure (WBM) and barley (Hordeum vulgare L.) straw-bedded manure (SBM). There were no significant differences in temperature regimes of SBM and WBM, indicating similar rates of successful composting. Of 17 physical and chemical parameters, five showed significant (P < 0.10) differences due to bedding at the outset of composting (Day 0), and 11 showed significant differences at final sampling (Day 124). During composting (10 sampling times), seven parameters showed significant bedding effects, 16 showed significant time effects, and four showed a Bedding x Time interaction. Significantly lower (P < 0.10) losses of nitrogen (N) occurred with WBM (19%) compared with SBM (34%), which has positive implications for air quality and use as a soil amendment. Other advantages of WBM compost included significantly higher total C (333 vs. 210 kg Mg(-1) for SBM) and inorganic N (1.3 vs. 1.0 kg Mg(-1) for SBM) and significantly lower total phosphorus (4.5 vs. 5.3 kg Mg(-1) for SBM). Our results showed that wood chip bedding should not be a problem for subsequent composting of the manure after pen cleaning. In combination with other benefits, our findings should encourage the adoption of wood chips over straw as a bedding choice for southern Alberta feedlots. PMID- 18396562 TI - Re: too young to kill? U.S. Supreme Court treads a dangerous path in Roper v. Simmons. PMID- 18396563 TI - Re: common pitfalls in the evaluation of testamentary capacity. PMID- 18396564 TI - Impact of erythrocyte Duffy antigen genetic polymorphism on the distribution of GroBeta-T, a novel human CXC chemokine. AB - PURPOSE: Grobeta-T, a human CXC chemokine, has been studied for its potential to mobilize stem cells. Chemokines bind specifically to receptors on target immune cells but also to a homologous erythrocyte blood group antigen, the Duffy Antigen/Receptor for Chemokines (DARC)that is subject to genetic polymorphism in humans.A mutation in the DARC gene is common among African Americans and results in lack of expression of the erythrocyte antigen. We used a combination of in vitro studies of Grobeta-DARC interaction and pharmacokinetic simulation to anticipate the potential impact of this polymorphism on the pharmacokinetics of Grobeta-T. METHODS: [125I]Grobeta-T was incubated in Caucasian blood to characterize the concentration dependence of the blood to plasma concentration ratio (B/P). Affinity and capacity of binding was estimated by Scatchard analysis; specificity was investigated by competitive displacement with a CC chemokine. The B/P value (7 nM) was then determined in blood from 8 African American subjects. Duffy antigen expression was determined by antibody agglutination. A pharmacokinetic model was developed which accounted for blood cell binding. Simulations were performed to explore effects of dose regimen and DARC expression on the Grobeta-T plasma concentration-time profile. RESULTS: Grobeta-T affinity and capacity for DARC (Caucasian blood)were 23.0 +/- 1.2 and 37.7 +/- 0.6 nM, respectively;excess CC chemokine fully displaced [125I]Grobeta T. Chemokine binding was highly correlated with the presence or absence of the Duffy antigen (p<0.01) in African American blood; the proportion of subjects for which binding was observed (3/8), was consistent with the reported frequency of DARC expression in this population. Counter to intuition,in the terminal disposition phase at low doses,concentrations of free Grobeta-T in the presence of DARC may be substantially higher than in the absence of DARC. CONCLUSION: Dissociation from the erythrocyte antigen may lead to greater persistence, at low doses, of free Grobetabeta-T in the blood of individuals expressing the chemokine sink. PMID- 18396567 TI - Studying nursing practice. PMID- 18396568 TI - [The creation of a leisure press by the pioneers of the middle class: Lyons 1870 1914]. PMID- 18396569 TI - ["A ride, mister?" Cabbies and the Lyons bourgeoisie of the Belle Epoque]. PMID- 18396570 TI - European perceptions of American coeducation, 1865-1914: ethnicity, religion and culture. PMID- 18396571 TI - Relief, restoration and reform: economic upturn yields modest and uneven health returns. AB - The sensitivity of state budgets to economic cycles contributes to fluctuations in health coverage, eligibility, benefits and provider payment levels in public programs, as well as support for safety net hospitals and community health centers (CHCs). The aftershocks of the 2001 recession on state budgets were felt well into 2004. More recently, the economic recovery allowed many states to restore cuts and, in some cases, expand health services for low-income people, according to findings from the Center for Studying Health System Change's (HSC) 2007 site visits to 12 nationally representative metropolitan communities. Along with bolstering support of safety net providers and raising Medicaid payments for private physicians, some states advanced even more ambitious health reform proposals. Yet across communities, safety net systems face mounting challenges of caring for more uninsured patients, and these pressures will likely increase given the current economic downturn. PMID- 18396572 TI - Equity in maritime health and safety--presentation of the 9th International Symposium on Maritime Health, Esbjerg, Denmark, 3-6 June 2007-12-19. PMID- 18396573 TI - The XXII World Congress of Apostleship of the Sea--the approach to the old and new challenges at maritime work-site. PMID- 18396574 TI - Fifth European meeting of the WHO collaborating centres in occupational health. Buxton, United Kingdom, 14-16 March 2007. PMID- 18396575 TI - 33rd Annual scientific meeting of the EUBS. 8-15 September 2007 Sharm El Sheik, Egypt. PMID- 18396577 TI - Maritime medicine days in Bergen 2007. PMID- 18396576 TI - SIRC symposium, Cardiff, 4-5 July 2007. PMID- 18396578 TI - Profiles of behavioral problems in children who witness domestic violence. AB - Unlike previous investigations of shelter-based samples, our study examined whether profiles of adjustment problems occurred in a community-program-based sample of 175 school-aged children exposed to domestic violence. Cluster analysis revealed three stable profiles/clusters. The largest cluster (69%) consisted of children below clinical thresholds for any internalizing or externalizing problem. Children in the next largest cluster (18%) were characterized as having externalizing problems with or without internalizing problems. The smallest cluster (13%) consisted of children with internalizing problems only. Comparison across demographic and violence characteristics revealed that the profiles differed by child gender, mother's education, child's lifetime exposure to violence, and aspects of the event precipitating contact with the community program. Clinical and future research implications of study findings are discussed. PMID- 18396580 TI - Self-regulation and the etiology of sexual deviance: evaluating causal theory. AB - Recently introduced theories of sex offending, including the self-regulation model and the multimodal self-regulation theory, have implicated self-regulatory deficits as a key variable in the development of sexually inappropriate interests and behaviors. While dysregulation has been considered an important component of a variety of behavioral, emotional, and interpersonal disorders, sexual behaviors have rarely been conceptualized within this context. In this study, we have examined a number of variables linked to self-regulation and dysfunctional outcomes in a sample of 95 sex-offending men. Results of a path analysis demonstrated that self-regulatory deficits were significantly predictive of paraphilic and antisocial behaviors in this group. Implications for our understanding of the etiology of sexual deviance and future research in this area are discussed. PMID- 18396581 TI - Attachment styles, alcohol, and childhood experiences of abuse: an analysis of physical violence in dating couples. AB - This study examined individual and partner characteristics that contribute to the propensity for physical violence in couples. In a sample of 171 heterosexual dating couples, each partner completed measures assessing experienced childhood abuse, alcohol use, alcohol expectancies, attachment, and relationship length. Physically violent men reported more abuse from each parent, greater alcohol use, anxious attachment, and a longer relationship. Their female partner reported more childhood abuse by the father and reciprocal perpetrated violence. Physically violent women reported more abuse from the father, greater alcohol use, aggressive alcohol expectancies, and a longer relationship. Their male partner reported greater abuse from the mother, greater alcohol use, and reciprocal perpetrated violence. This study demonstrates the importance of considering how each individual's characteristics within a dyad contribute to increased propensity for dating violence. PMID- 18396579 TI - Trajectories of physical and emotional marital aggression in midlife couples. AB - This study used three-level generalized hierarchical linear modeling to examine trajectories of husbands' and wives' physical and emotional aggression over three assessments and the effects of years since marriage. In this community sample of 118 couples, physical aggression significantly decreased over time (43% per year). Emotional aggression did not significantly change over time, but trajectories significantly differed for husbands (3% increase) versus wives (10% decrease). Longer-duration marriages had lower physical aggression and, for wives only, lower emotional aggression. Aggression trajectories showed considerable variability: 44%-55% of physically aggressive spouses desisted from one assessment to the next; 5%-12% reported start-ups in physical aggression. Discussion addresses the role of gender and type of aggression in aggression trajectories. PMID- 18396582 TI - A test of explanations for the effect of harsh parenting on the perpetration of dating violence and sexual coercion among college males. AB - This study uses structural equation modeling (SEM) with a sample of 760 college males to test various hypotheses regarding the avenues whereby harsh corporal punishment and a troubled relationship with parents increase the risk that a boy will grow up to engage in sexual coercion and dating violence. We found that three variables--a general antisocial orientation, sexually permissive attitudes, and believing that violence is a legitimate component of romantic relationships- mediated most of the association between negative parenting and our two outcomes. In addition to this indirect influence, we found that harsh corporal punishment had a direct effect upon dating violence. The findings are discussed with regard to various theoretical perspectives regarding the manner in which family of origin experiences increase the chances that a young man will direct violence toward a romantic partner. PMID- 18396583 TI - Measurement and correlates of prosocial bystander behavior: the case of interpersonal violence. AB - The field of social psychology has long investigated the role of prosocial bystanders in assisting crime victims and helping in emergency situations. This research has usually been experimental and has established important principles about the conditions under which individuals will choose to engage in prosocial bystander behaviors. More recently, interest has grown in applying this work to the important practical problem of preventing interpersonal violence in communities. Yet, to date, there has been little research on the role of bystanders in cases of interpersonal violence. The current study is thus exploratory. Using a sample of 389 undergraduates, the study discusses key issues in the development of measures to investigate these questions and presents preliminary analyses of correlates of bystander behavior in the context of sexual and intimate partner violence. PMID- 18396584 TI - Intimate partner homicide methods in heterosexual, gay, and lesbian relationships. AB - Previous research indicates that the killing method used in homicides may reflect the motivation of the offender and qualities of the victim-offender relationship. The effect of gender and sexual orientation of intimate partner homicide offenders (N = 51,007) was examined with respect to the brutality of killing methods. Guided by previous research and theory, it was hypothesized that homicide brutality will vary with the offender's sexual orientation and gender, such that the percentage of killings coded as brutal will be higher for (a) gay and lesbian relative to heterosexual relations, (b) men relative to women, (c) gay relative to heterosexual men, and (d) lesbian relative to heterosexual women. The rates of intimate partner homicide were also hypothesized to vary with the gender of the partners, such that (a) homicide rates will be higher in gay relative to heterosexual and lesbian couples and (b) homicide rates will be lowest in lesbian couples. The results support all but one prediction derived from the two hypotheses. We predicted that men would kill their partners more brutally than would women, but the results indicate that the opposite is true. PMID- 18396585 TI - Gender differences and socioeconomic status biases in judgments about blame in date rape scenarios. AB - Participants (80 men, 80 women) read hypothetical date rape scenarios, wherein the perpetrator's socioeconomic status (bus driver versus doctor) and the victim's level of resistance (verbal versus verbal and physical) were varied, and made judgments about who was at fault and what the consequences should be. In general, men assigned more blame to the victim and less blame to the perpetrator than did women. However, men assigned more blame to the bus driver than to the doctor. Women, on the other hand, assigned more blame to the victim who was raped by the bus driver than to the victim who was raped by the doctor. The results also indicated that participants recommended harsher punishments for the perpetrator when the victim resisted verbally than when she resisted verbally and physically. Future research on the role of the perpetrator's, the victim's, and the participants' socioeconomic status in judgments about date rape is suggested. PMID- 18396587 TI - Warding off sleepiness. AB - This article focused on the sleep effects of three schedule attributes. Ideally, business requirements and employee preferences also will be taken into account when selecting the "best" shift schedule for your site. PMID- 18396588 TI - Tour of duty. PMID- 18396589 TI - Cover photo lacked proper rope rescue techniques. PMID- 18396590 TI - We're going back to Vegas, baby. PMID- 18396591 TI - At home, work, or play, it's hot. PMID- 18396592 TI - Beware the sliding program. PMID- 18396593 TI - Monitoring keeps odors, dust, particles at bay. PMID- 18396594 TI - Safety and health incentives: a holistic approach. PMID- 18396595 TI - Enhancing safety before breaking ground. PMID- 18396596 TI - Send the protection message loud and clear! PMID- 18396597 TI - If I had a hammer, I'd wear hearing protection all day long. PMID- 18396598 TI - Rethinking rig embarkation. PMID- 18396599 TI - Recognizing and overcoming barriers to colorectal screening in primary care. AB - Colorectal cancer screening represents a major health issue that can be improved at the primary care level by individualized, focused attention to screening. We describe a practice-based method to increase awareness of the potential barriers to effective screening and our attempts to overcome these barriers. A recent study indicating the prevalence of colorectal neoplasm in patients with documented coronary artery disease emphasizes the pivotal role that primary care providers play in screening procedures across the medical spectrum. While we are intuitively aware of increased screening success, we intend to document the change in our effectiveness since we initiated this effort in January 2004. PMID- 18396600 TI - Bancroftian filariasis in Charleston, South Carolina: the medical history of a forgotten and abhorred disease. PMID- 18396602 TI - Using our brains. PMID- 18396601 TI - Long-term survival following salvage reirradiation with concurrent chemotherapy for recurrent head and neck cancer. PMID- 18396603 TI - It's the law: physicians must report abuse. PMID- 18396604 TI - The Bard and Liberty. PMID- 18396605 TI - A walking tour of Charleston--medical history. PMID- 18396606 TI - Medical research: carving the future health care in Papua New Guinea. History of the Papua New Guinea Medical Journal and the Medical Society of Papua New Guinea. PMID- 18396607 TI - A qualitative evaluation of the immunization program in Papua New Guinea. AB - OBJECTIVE: An in-depth evaluation of rural immunization services in Papua New Guinea was conducted to determine the reasons for poor immunization coverage, as well as to document the impact of recent efforts to strengthen the national immunization program. METHOD: A qualitative process was used to complement quantitative monitoring data. An interview process, based on open-ended questions, active listening and observation, was designed whereby a team of program supervisors collected information from rural health staff. The teams interviewed health staff in 30 health centres that were selected to provide examples of contrasting field situations. RESULTS: This qualitative review provided valuable detail about why immunization services were failing, encompassing locally specific weaknesses, such as logistic reasons for not conducting outreach, and generic systemic problems such as lack of access to funding. In addition, the information gathered provided details on local solutions developed by better-performing facilities. Both these aspects added significant value to quantitative measures of program performance (derived from national health information system data and analysis of supervision checklists). The review also captured a number of behavioural reasons that will need to be overcome before an improvement in the services can be expected. CONCLUSIONS AND IMPLICATIONS: This in-depth evaluation provided valuable information about problems in peripheral immunization clinics and identified local solutions. The high level of detail collected will be important for planning future strengthening of the health system. The study modelled a supportive form of supervision with the potential to improve outcomes from future supervisory visits. Some of the major barriers to improved immunization were locally specific organizational issues, as well as complex human problems. While some issues can be remedied through further strengthening of immunization systems, others lack easy, rapid solutions. PMID- 18396608 TI - Impact of pre-packaging antimalarial drugs and counselling on compliance with malaria treatment at Port Moresby General Hospital Adult Outpatient Department. AB - We investigated the impact of pre-packaging antimalarial drugs and counselling on compliance with treatment of malaria at the Adult Outpatient Department of Port Moresby General Hospital. Adult patients who were prescribed standard antimalarial drugs following clinical and microscopic diagnosis of malaria were randomly assigned to one of three groups: an intervention group, where pre packaging and counselling instructions were applied; control group A, with counselling but no pre-packaging; and control group B, with neither counselling nor pre-packaging. Patients were interviewed on two occasions, day 1 of treatment and day 4 post treatment. Of a total of 436 patients, 322 patients (179 males and 143 females) completed the study. Our data indicate an increase of 18% in compliance with treatment in the intervention group and 16% in control group A, when compared with control group B. While compliance with treatment was gender independent, the language spoken and used for giving instructions and counselling may have influenced patients' behaviour on prescribed medication. The results of our study indicate that a simple pre-packaging system and proper counselling could improve compliance with antimalarial drug treatment. As an additional beneficial observation, pre-packaging is likely to eliminate errors and possible contamination of the products during dispensing. PMID- 18396609 TI - Cross-sectional characterization of malaria in Sanma and Shefa Provinces, Republic of Vanuatu: malaria control implications. AB - Endemic malaria still exists in the Republic of Vanuatu, an 80-island archipelago that sits astride the southeast margin of the Southeast Asian-Melanesian malaria band (Buxton Line 170 degrees E, 20 degrees S). The annual parasite incidence has decreased dramatically over the past decade, which has been attributed to an intensive insecticide-treated bednet distribution program and implementation of a revised Plasmodium falciparum treatment policy that employs combination chloroquine + sulfadoxinelpyrimethamine as a first-line therapy. Standard malariometric surveys were conducted at 10 locations in 2 provinces, screening 2351 adults and children towards the end of the peak transmission season. Spleen rates were consistent with mesoendemic malaria. Examination of blood slides revealed a mean slide-positive rate of 22% (range 4% to 33%). P. falciparum predominated, accounting for 73% of infections, followed by P. vivax (25%). Among 396 individuals with P. falciparum, the gametocyte rate was 54%, with 37% presenting gametocytes alone without asexual stages. Only 8% and 4% of persons with asexual stage P. falciparum and P. vivax parasitaemia, respectively, were symptomatic. These data suggest that malaria transmission has increased in some locations in Vanuatu over the past decade and this report underscores the importance of appropriate bednet use and vector control in this setting as well as the impact of adding sulfadoxine/pyrimethamine and removing primaquine from the national malaria treatment formulary. PMID- 18396610 TI - The challenges to scaling up antiretroviral therapy in Papua New Guinea. AB - This study used a qualitative interview methodology to explore the barriers to scaling up access to antiretroviral therapy (ART) in Papua New Guinea (PNG). Key informants were selected purposively from organizations active in HIV/AIDS (human immunodeficiency virus/acquired immune deficiency syndrome) work throughout PNG. Interviews were taped, transcribed and analyzed thematically. Predicted barriers likely to impede the delivery of ART by the health services included shortages of skilled and experienced personnel; lack of sustainable funding; weak services at the rural periphery; lack of capacity for the management of chronic illness; inadequacies of basic services for people living with HIV/AIDS (PLWHAs); the difficulty of finding the most appropriate method and venue of delivery for ART; and the complexities of establishing social criteria for deciding who to treat. Barriers likely to impede the acceptance of treatment by the community included the cost of treatment to the patient; the low socioeconomic status of PLWHAs; the effects of stigma, particularly in the case of women; and sociocultural factors affecting public perceptions of ART. The study concluded that, given the range of potential barriers, scaling up ART in PNG and other developing countries will be a complex process, requiring careful decisions to be made over how best to utilize available resources. PMID- 18396611 TI - A serological survey of rickettsial infections in the Gazelle Peninsula, East New Britain and a review of the literature. AB - Of the various rickettsial diseases, only scrub typhus has been well documented in Papua New Guinea. A review of the historical literature confirms this. A serological survey was conducted on 113 antenatal patients presenting to a district hospital in Kokopo, East New Britain. Results suggested that a spotted fever rickettsial infection is common in this area with a seroprevalence of about 17% in young women. There was no evidence of scrub typhus or murine (endemic) typhus in the population sampled. Clinical implications of these findings are discussed. PMID- 18396612 TI - The effect of betel quid chewing on the Achilles tendon reflex time. AB - This study investigated the effect of betel quid chewing on the somatic motor pathway (Achilles tendon reflex time). The subjects, 48 healthy male volunteers, were divided into 4 groups: a control group; first-time chewers; occasional chewers; and habitual chewers. The Achilles reflex time (ART) of the subjects was taken 5 minutes after ingestion of the quid preparation and their pulse rate and respiratory rate were also recorded. The results show that betel quid chewing reduced the ART and increased the pulse rate and respiratory rate. It can be concluded that betel quid chewing causes a transient peripheral stimulation of the somatic motor pathway with a resultant hyperreflexia. PMID- 18396613 TI - Cannabis in Papua New Guinea. AB - Cannabis is the most commonly used illicit drug in Papua New Guinea (PNG). Data on the epidemiology and history of cannabis in PNG are presented. The adverse health consequences of cannabis smoking and treatment options for cannabis dependence are discussed. It is recommended that a range of strategies are urgently required in PNG to prevent adverse physical and mental health consequences associated with cannabis smoking. PMID- 18396614 TI - Galbulimima bark and ethnomedicine in Papua New Guinea. AB - This article reviews the literature and current information on the use of Galbulimima bark in traditional medicine in Papua New Guinea. Galbulimima bark is used in Papua New Guinea as an analgesic, to treat fever or to get rid of head lice. It is used in divination to produce trance-like states for counteracting malevolent power that is thought to be the cause of a variety of illnesses. Galbulimima bark has also been chewed, or drunk as a decoction, to induce visions and a dream-like state. It is also chewed and then rubbed on the legs before fighting. PMID- 18396615 TI - [To understand the principles and characteristics of multi-slice computed tomography and enhance its clinical applications]. PMID- 18396616 TI - [Diagnostic accuracy of noninvasive coronary artery angiography using 64 multi slice computed tomography on coronary atherosclerosis]. AB - OBJECTIVE: To evaluate the diagnostic accuracy of 64 multislice computed tomography (MSCT) coronary angiography on the coronary atherosclerosis. METHODS: Sixty one (52 male, 9 female, averaged 58 +/- 11 yrs ) patients underwent conventional coronary angiography and 64-MSCT angiography for suspected coronary artery disease within 5 to 20 days. Coronary artery image quality was scored from 1 to 4 (image quality score, 1: poor; 2: good -; 3: good +, and 4: excellent) on RCA, LM, LAD and LCX, respectively. The coronary artery stenosis were divided into two groups by over than 50% and less than 50% and diagnostic accuracy was analyzed. The coronary plaques were divided as calcified and noncalcified plaque according to its density. The stenosis result from calcified and noncalcified plaque were analyzed. RESULTS: Mean score of image quality on RCA, LM, LAD and LCX was 3.57 +/- 0.18 with heart beats 50 - 104 bpm (mean 70 +/- 11 bpm). Compared with CAG, the sensitivity and specificity of 64-MSCT in evaluating stenosis are 94.45% and 96.15%, the false positive and false negative value are 3.85% and 4.55%. The sensitivity and specificity were 85.71% in evaluating stenosis result from noncalcified plaque and they were 83.33% and 31.57% result from calcified plaque. CONCLUSION: 64-MSCT is an accurate and feasible method in evaluating coronary artery stenosis compared to conventional coronary angiography. However the specificity was poor in evaluating calcified plaque. PMID- 18396617 TI - [Value of multi-slice computed tomography in diagnosis of coronary plaque characterization]. AB - OBJECTIVE: To investigate the value of multi-slice computed tomography in diagnosis of different types of coronary atherosclerotic plaques. METHODS: Twenty eight patients undergoing CT angiography (CTA) with normal coronary arteries were randomly selected to measure the CT values of different sections of the 4 main branches of coronary artery. Twenty-five specimens of human heart from the bodies of the patients who died of non-cardiogenic diseases were scanned by 16-slice CT scanner and 64-slice CT scanner: a mixture of CT contrast media and normal saline was injected into the coronary arteries to achieve in-vivo-like contrast enhancement within the coronary artery lumen to detect atherosclerotic plaques. The CT values of plaques were measured in several regions of interest (ROI) selected in each plaque. The CT images thus obtained were evaluated by 2 experienced radiologists. There are nine specimens with coronary atherosclerotic plaques among them. Then the atherosclerotic lesions in the coronary were made into tissue specimens to undergo pathological examination. RESULTS: 7560 CT values were obtained from the 28 patients. Thirty-eight atherosclerotic plaques were found by CAT in 9 heart specimens and confirmed by pathology. When the CT value of coronary lumen was 370 HU, the predominant lipid-rich plaque showed a mean CT value of 53 +/- 12 HU; the fibrous-rich plaque showed a mean CT value of 106 +/- 17 HU; and the calcified plaque showed a mean CT value of 429 +/- 94 HU measured by 16-slice CT; and the predominant lipid-rich plaque showed a mean CT value of 51 +/- 13 HU; the fibrous-rich plaque showed a mean CT value of 110 +/- 19 HU; and the calcified plaque showed a mean CT value of 435 +/- 87 HU measured by 64-slice CT. The CT value of the fibrous-rich plaque was significantly higher than that of the lipid-rich plaque (P = 0.008), and lower than that of the calcified plaque (P < 0.01). There was no significant difference between the results obtained by the two kinds of CT scanners. CONCLUSION: CTA can non invasively assess the atherosclerotic plaques. PMID- 18396618 TI - [The role of multi-detector row CT in the diagnosis and hemodynamic studies of gastric varices in portal hypertension]. AB - OBJECTIVE: To evaluate the value of multi-detector row CT (MDCT) in the diagnosis and hemodynamic studies of gastric varices (GV) in portal hypertension by comparison with endoscopy and DSA direct portography. METHODS: Thirty-six consecutive cirrhotic patients with GV confirmed by endoscopy underwent tri-phase contrast-enhanced CT scans and CT portography (CTP) within 2 weeks after endoscopy examination. Three independent experienced radiologists, who were blinded to the patients' clinical data, analyzed the CT images, including the size and location of GV as well as afferent and efferent veins of GV, separately. Interobserver agreement among the 3 radiologists with regard to the diagnosis of submucosal and perigastric GV was determined by Kappa (k) values. The findings of endoscopy were used as standards. RESULTS: Sub mucosal GV was diagnosed in 34 of the 36 patients (94.4%) and perigastric GV in all 36 patients (100%) by the observation of the 3 radiologists. MDCT showed an excellent interobserver reliability with regard to the diagnosis of submucosal GV (kappa = 0.85) and perigastric GV (kappa = 1.0). Agreement between MDCT and endoscopy with regard to the opacification of variceal size and location were 86.1% and 88.9% respectively. The sensitivity, specificity, accuracy, and positive predictive value of CTP in the opacification of afferent and efferent veins of GV were all more than 80%. The frequencies of participation of posterior gastric vein and short gastric vein in blood supply to gastric fundal varices in the isolated gastric varices and gastroesophageal varices type 2 (GEV2) were 94.1% and 70.6% respectively, both significantly higher than those in the gastroesophageal varices type 1 (GEV1, 52.6% and 31.6%, respectively, both P < 0.05). The main blood drainage route of GEV1 was via azygous system into the super vena cava (100%), whereas in the gastric fundal varices the main blood drainage route was via the gastrorenal shunts into the inferior vena cava (82.4%). CONCLUSION: MDCT can be used as an important tool for detecting submucosal and perigastric GV, and can clearly reveal the size, location, and hemodynamics of GV. PMID- 18396619 TI - [Features of obturator nerve by high-resolution CT at the same slice with multiplanar planar reconstruction techniques and clinic value thereof]. AB - OBJECTIVE: To explore the features of obturator nerve (ON) by high-resolution CT (HRCT) at the same slice with multiplanar planar reconstruction techniques and clinic value thereof. METHODS: The normal lumbar vertebral bodies of 60 healthy adults and the abnormal lumbar vertebral bodies of 31 patients with ON disorders were scanned with 16-slice spiral CT, multiplanar reconstruction was conducted in the Workstation ADW4. 1, with UNIX system and DICOM form. The images at the same slice were studied. RESULTS: The shapes, thickness, direction, length, adjourning relationship, and tension could be shown at the same slice of oblique and coronal section or oblique and sagittal plane with rotation in external sides on the way from the starting point of dural sac to foramen obturatum in pelvis, meanwhile the manifestations of abnormal ON also could be shown. CONCLUSION: The anatomical features and abnormal manifestation of ON at the same slice with 16-slice spiral CT using the multiplanar reconstruction techniques can be fully shown from the starting point of dural sac to the foramen obturatum in pelvis. This technique is very important in diagnosis and treatment of diseases of obturator nerve. PMID- 18396620 TI - [Correlation between serum uric acid and insulin resistance]. AB - OBJECTIVE: To assess the correlation between serum uric acid and insulin resistance in natural population of Beijing. METHODS: 1 056 people without diabetes, 460 males and 596 females, aged 35-64 was selected from the natural population in Beijing using stratified-random sampling method. Serum uric acid, fasting blood glucose, insulin and other metabolic parameters were measured. The insulin activity index (IAI) was calculated to assess insulin resistance. RESULTS: The natural logarithm of IAI was decreased with the elevation of serum uric acid level (F = 47.884, P < 0.001). The prevalence of insulin resistance increased with the elevation of uric acid level, from 13.8% in quartile 1 to 33.9% in quartile 4. Compared to the quartile 1, the odds ratio (OR) of the quartile 2 for prevalence of insulin resistance was 1.935; the OR of the quartile 3 was 1.590; the OR of the quartile 4 was 2. 088. Uric acid was independently correlated with insulin resistance after adjustment for sex, age and main components of metabolic syndrome (MS). CONCLUSION: Serum uric acid is correlated with insulin resistance in Beijing natural population after adjustment of sex, age, and the main components of MS. PMID- 18396621 TI - [Removal and plastic reconstructive surgery of orbital neurofibroma]. AB - OBJECTIVE: To investigate the safety and efficacy of surgical management of orbital neurofibroma (NF). METHODS: Twenty-six patients with orbitotemporal NF underwent tumor resection. And then 12 of the 26 patients that were of Jackson group I, underwent blepharoplasty surgery, including lid plastic and reconstructive surgery, lateral canthal reattachment and ptosis surgery. Ten patients of group II underwent blepharoplasty combined with orbital reconstruction. And 4 patients of group III underwent blepharoplasty surgery, orbital reconstruction, and hydroxyapatite orbital implanting. The functionally useless eyes of 2 patients were retained for cosmetic appearance. RESULTS: There was no recrudescence after neurofibroma resection. 10 patients had their vision acuity improved. All patients obtained satisfying results after eyelid and orbital plastic surgery. 2 patients relapsed into ptosis that was cured with further frontalis suspension procedure with autogenous fascia lata. CONCLUSION: Neurofibroma removal surgery is a safe and effective treatment method for NF patients. The periorbital appearance and visual acuity of patients with orbitotemporal NF can be significantly improved through oculoplastic surgery. PMID- 18396623 TI - [CT guided radioactive 125I seed implantation in treating localized advanced pulmonary carcinoma]. AB - OBJECTIVE: To investigate the clinical value of CT guided radioactive 125I seed implantation (CTRISI) in the treatment of localized advanced pulmonary carcinoma. METHODS: Thirty-two biopsy -confirmed localized advanced pulmonary carcinoma patients (Group A) with 48 lesions, 5.5 (4.5 -7.5) cm in diameter, clearly demarcated from the adjacent normal tissue, who failed to be improved in the first line chemotherapy, underwent CI-guided implantation of 125I seeds of 23.3 - 30.0 megabecquerel (MBq) into these lesions with the matched peripheral dose of 100-150 Gy. Thirty patients (Group B) treated by first line chemotherapy were randomly selected as the control group. RESULTS: CT follow-up two months later proved that the local control rate of Group A was 78.1%, significantly higher than that of Group B; the one year survival rate of Group A was 65.0%, significantly higher than that of Group B (48.0%); the median survival time of Group A was 15 months, significantly longer than that of Group B (11 months); no myelosuppression was found in Group, and the myelosuppression rate of Group B was 46.3%. In Group A small amount of effusion was observed, pneumothorax occurred in 4 cases with one of the lungs compressed by less than 30% that was improved after conservative treatment; one week after the procedure bloody sputum occurred in 15 cases. CONCLUSION: Improving the living quality obviously and with good short term effects and few complications, CTGRISI procedure is safe and well-tolerated in treating localized advanced pulmonary carcinoma in Chinese patients. PMID- 18396622 TI - [Correlation of Her-2 overexpression with endocrine status and response to tamoxifen treatment in patients with breast cancer]. AB - OBJECTIVE: To investigate the correlation of Her-2 overexpression with endocrine status and response to tamoxifen treatment in patients with breast cancer. METHODS: Immunohistochemistry was used to measure the expression of estrogen receptor (ER), progesterone receptor (PR), Her-2, p53, proliferating cell nuclear antigen (PCNA) in 4773 consecutive in-hospitalized patients with primary breast cancer confirmed by pathological examination of the resected specimens, all females, aged 50 (15 - 92), and were followed up for 42 months on average. 1090 if then received tamoxifen (TAM), an anti-estrogen agent, post-operationally. The correlation of Her-2 overexpression with other factors was analyzed. RESULTS: (1) The Her-2 overexpression rate was 26.6%. Her-2 overexpression was significantly negatively correlated with expressions of ER and PR (both P < 0.01). (2) The general 3-year disease-free survival (DFS) rate of those treated with TAM was 92.7%; and the 3-year GFS rate of the subgroup of the TAM-treated patients with Her-1 overexpression was 91.2%, significantly lower than that of those without Her-2 overexpression (93.4%, P = 0.004). (3) The 3-year DFS rate premenopausal patients with Her-2 overexpression was 91.8%, not significantly different from that of those without Her-2 overexpression (92.6%, P > 0.05), however, the 3-year DFS rate of the postmenopausal patients with Her-2 overexpression was 90.4%, significantly lower than that of those without Her-2 overexpression (92.6%, P = 0.010). (4) The 3-year DFS rate of the axillary lymph node-positive patients with Her-2 overexpression was 89.1%, significantly lower than that without Her-2 overexpression (92.3%, P = 0.037). (5) In the premenopausal patients there was no significant difference in the 3-year DFS rate between the lymph node-negative patients with and without Her-2 overexpression (P > 0.05). However, in the postmenopausal lymph node positive patients the 3-year DFS rate of those with Her 2 overexpression was 88.7%, significantly lower than that of those without Her-2 overexpression (92.2%, P = 0.0069). CONCLUSION: ER and PR are independent factors of Her-2 expression. Her-2 overexpression signifies resistance to TAM treatment. The response to TAM is not influenced by the Her-2 expression and axillary lymph node metastasis status in the premenopausal patients, and is not influenced by the Her-2 expression level in the postmenopausal patients. PMID- 18396624 TI - [Diagnostic and follow up value of magnetic resonance cholangiography in biliary complications after orthotopic liver transplantation]. AB - OBJECTIVE: To evaluate the diagnostic and follow up value of magnetic resonance cholangiography (MRC) in biliary complications after orthotopic liver transplantation (OLT). METHODS: MRC was performed, using 1.5 T scanner on 166 consecutive OLT recipients. The MRC results were analyzed and compared with the confirmed diagnosis criteria (direct cholangiography, laparotomy, and liver puncture). RESULTS: The sensitivity, specificity, accuracy, and positive and negative predictive value of MRC in diagnosis of biliary complications were 98.4%, 81.6%, 87.9%, 76.5% and 98.8%. 104 clinically normal cases and 62 symptomatic cases showed 31 and 50 MRC abnormal findings respectively. MRC totally showed 70 cases of suspected biliary complications following OLT, including 41 cases with anastomotic stricture, 8 with focal non-anastomotic stricture, and 12 with stones or debris formation, 2 with bile leak, 3 with cholangitis and 4 with oppressive changes of biliary tract. MRC also showed periportal edema in 11 cases. CONCLUSION: A good imaging modality to show both periportal edema and biliary abnormality of various patterns, MRC provides important information for post-operational follow up and early detection of biliary complications in OLT recipients. PMID- 18396625 TI - [Clinical analysis of late-onset nephrotic syndrome after allogenic hematopoietic stem cell transplantation]. AB - OBJECTIVE: To investigate the clinical manifestations, pathology, diagnosis, treatment, and pathogenesis of late-onset nephrotic syndrome (NS) after allogenic hematopoietic stem cell transplantation (allo-HSCT). METHODS: NS post-HSCT was investigated in 167 patients with hematopoietic malignancies who survived more than 3 months after allo-HSCT. The clinical manifestations, pathology, diagnosis, and treatment were investigated in a retrospective study. The association of the onset of NS post-HSCT with sex, age, transplant type, conditioning regiments, human leucocyte antigen (HLA) matching, donor-recipient relationship, acute graft versus host disease (aGVHD), chronic graft versus host disease (cGVHD), and cytomegalovirus infection were analyzed. RESULTS: Five patients (2.99%) were diagnosed as with NS post-HSCT, 4 with membranous glomerulonephritis (MGN) and 1 with minimal change disease (MCD). Immunohistochemistry of glomerular lesions revealed that the immunoglobulin (Ig) of immune complex deposition included IgG in 3 patients, IgM in 1 patient, and co-existence of IgG/IgM in 1 patient. Serum ANA was positive in 2 patients and serum IgG and IgM were in high levels in 2 patients. Glucocorticoid combined with cyclophosphamide (CTX) was fundamentally effective treatment for NS post-HSCT. 1 patient got complete response, 3 got partial response, and 1 was stable after the treatment. Binary logistic regression showed that the he onset of NS post-HSCT was not significantly associated with sex, age, transplant type, conditioning regimen, HLA matching, donor-recipient relation, aGVHD, cGVHD and CMV infection. CONCLUSION: The predominant pathological type of NS post-HSCT is MGN, followed by MCD. The pathogenesis of NS post-HSCT may be related to the abnormality of humoral immunity. Glucocorticoid combined with CTX is an effective treatment. PMID- 18396626 TI - [Relationship of brain natriuretic peptide to renal function and left ventricular hypertrophy in patients with chronic kidney disease]. AB - OBJECTIVE: To clarify the association of brain natriuretic peptide * BNP) with renal function and cardiac dysfunction in non-dialysis-dependent patients with chronic kidney disease (CKD). METHODS: Fluorescence immuno-assay was used to detect the whole blood concentration of BNP in a cohort of 203 CKD patients and 16 hypertensive controls. Color ultrasonography was conducted to determine the left ventricular mass index (LVMI). Estimated glomerular filtration rate (eGFR) was examined. The relationship of BNP to eGFR and LVMI in these patients. RESULTS: There was a trend that the BNP level increased with the decrease of the level of eGFR [9.35 (7.35-15.00) vs 54.40 (15.10-173.00) ng/L, P < 0.01]. Spearman correlation showed that BNP level was correlated negatively with eGFR (r = -0.417, P < 0.01). Multiple regression analysis showed that eGFR was the independent determinant of BNP (beta = -0. 293, P < 0.01), and lgBNP concentration increased by 0.293 ng/L per 1 ml x min(-1) x (1.73 m2)(-1) reduction in lgGFR. The incidence of LV hypertrophy was much higher in the CKD patients than in the controls (53.2% vs 10.0%, P < 0.05). BNP level was greater in the patients with LV hypertrophy [93.05 (37.70-272.00) vs 17.30 (7.20-63.70) ng/L, P < 0.01], and LVMI had an independent effect on BNP concentration ( beta = 0.266, P < 0.01). lgBNP level increased by 0.266 ng/L per 1 g/m2 increase in lgLVMI. CONCLUSION: The declining eGFR level independently elevates the BNP level in non-dialysis-dependent CKD patients. Although renal dysfunction itself may affects the BNP concentration, LV hypertrophy is a powerful independent determinant of BNP. PMID- 18396627 TI - [Expression of long isoform leptin receptor and shortest membrane bound variant in peripheral blood mononuclear cells from the obese and normal individuals]. AB - OBJECTIVE: To investigate the expression of the long isoform leptin receptor (OB R(L)) and the shortest membrane bound variant (OB-R(S)) in peripheral blood mononuclear cells from the obese and normal individuals (6.08 ng/ml or 31.21 ng/ml). METHODS: Mononuclear cells were obtained from 30 obese individuals (BMI > 25 kg/m2) and 20 normal individuals (BMI < 23 kg/m2). RT-PCR was used to detect the expression of OB-R(S) and OB-R(L). The level of serum leptin was measured with immunoradiometric assay. RESULTS: OB-R(S) was expressed in all individuals, and OB-R(L) was expressed in 38 individuals. OB-R(L) was not expressed in the 12 obese individuals with the BMI > 39 kg/m2. The expression level of OB-R(S) was 4 times higher than that of OB-R(L). There was no significant difference in the expression of either isoforms between men and women. The relative expression of both OB-R(S) and OB-R(L) isoforms was significantly lower and the serum leptin level was significantly higher in the obese individuals (6.08 ng/ml or 31.21 ng/ml). The leptin receptor expression levels were significantly negatively correlated with BMI and serum leptin level. CONCLUSION: Both OB-R(S) and OB-R(L) are expressed in the peripheral blood mononuclear cells from normal and the medium obese subjects with a consistent predominance of OB-R(S). There is no significant difference in the expression of OB-R(S) and OB-R(L) between men and women. Compared with normal individuals, the expression of OB-R(S) and OB-R(L) in human mononuclear cells is lower. PMID- 18396628 TI - [HIV-1 genotypic resistance profiles in children failing highly active antiretroviral therapy]. AB - OBJECTIVE: To study the genotypic resistance profiles of HIV-1 children failing highly active antiretroviral therapy (HAART) so as to provide helpful information for the treatment regime of Chinese children infected with HIV-1. METHODS: Peripheral venous blood samples were collected from 20 HIV-1 infected children of Henan province, aged 9 (3 - 14). Nested RT-PCR was used to amplify part of the RT (40 -250 aa) gene. The PCR products of RT gene underwent nucleotide sequencing, the resulting nucleotide sequences were analyzed by the HIVdb data offered by the Stanford University web site to find the drug resistance mutations. RESULTS: (1) Phylogenetic analysis revealed that 20 of the RT sequences were classified as subtype B. (2) According to the genotypic analysis, 20 , 15, and 13 children showed high level resistance to the nevirapine. (NVP), delavirdine (DLV), and efavirenz (EFV) respectively; 7 and 5 children showed high and intermediate level resistance to azidothymidine (AZT) respectively. Five children showed potential low-level and intermediate level resistance to lamivudine (3TC), and 11 showed high level resistance to 3TC; 11 showed intermediate and high level resistance to stavudine (d4T) and didanoside (ddI) respectively; and 19 and 12 children showed resistance to abacavir (ABC) and tenofovir (TDF) which had never been taken by these children. CONCLUSION: The emergence of HIV resistant strains during antiretroviral therapy is one of the main reasons for treatment failure in HIV infected children. PMID- 18396629 TI - [Prospective, multicenter study of mortality and risk factors in children with acute respiratory distress syndrome in 25 pediatric intensive care units in China]. AB - OBJECTIVE: To investigate the mortality and predictors of outcome of children with acute respiratory distress syndrome (ARDS) in pediatric intensive care unit (PICU). METHODS: ARDS cases were selected from the 12 018 patients admitted in 25 pediatric intensive care units in China from January 1 to December 31, 2004, aged 29 days to 14 years, using the Chinese Pediatric Critical Index of Severity (PCIS) and American Guidelines for Admission and Discharge Policies for PICU. ARDS was diagnosed according to the 1994 American-European Consensus Conference criteria. RESULTS: 105 of the 12 018 patients (1.44%) were diagnosed as with ARDS. The overall mortality of ARDS was 61.0% (64/105), 9 times as high as that of the 7269 severe cases in PICU. Logistic regression analysis showed that infiltration shadows in 2 - 3 quadrants, pediatric critical illness score (PCIS), and partial pressure of carbon dioxide (PaCO2) at the onset of ARDS were independently associated with the mortality. CONCLUSION: ARDS has a high risk of death, and the infiltration shadows in 2-3 quadrants, PCIS, and PaCO2 are independently associated with mortality. PMID- 18396630 TI - [Antiatherosclerotic mechanism of aspirin: experiment with rabbits]. AB - OBJECTIVE: To observe the effects of aspirin on nuclear factor kappaB (NF-kappaB) DNA binding activity and on the expression of cyclooxygenase-2 (COX-2) in atherosclerotic plaque so as to explore its antiatherosclerotic mechanism. METHODS: Thirty-six New Zealand male rabbits were randomly divided into 3 equal groups: high-cholesterol (HC) group, fed with food high in cholesterol and perfused into the empty stomach daily with distilled water for 12 weeks, high cholesterol and aspirin (HC + A) group, fed with food high in cholesterol and perfused into the empty stomach daily with aspirin solution, and normal control (NC) group, fed with normal food and perfused into the empty stomach daily with distilled water, before the experiment, and 4, 8, and 12 weeks after the beginning of experiment peripheral blood samples were collected. The serum lipids were detected with enzymatic assays; and enzyme-linked immunosorbent assay was used to detect the level of high sensitive C-reactive protein (hs-CRP). By the end of experiment the rabbits were killed to take out the specimens of aorta to observe the neointima thickness and plaque area of aorta. Electrophoretic mobility shift assay was used to detect the NF-kappaB-DNA binding activity, and immunohistochemistry and morphometry were performed to observe the expression of COX-2 protein, the neointima thickness and plaque area of aorta respectively in all three groups. RESULTS: The levels of serum lipids, hs-CRP, NF-kappaB-DNA binding activity, expression of COX-2 protein, and neointima thickness and plaque area of aorta in the HC and HC + A groups were all significantly higher than those in the NC group (P < 0.05 -0.01). There was no significant differences in the serum lipids between the HC and HC + A groups (all P > 0.05), however, the levels of hs-CRP, NF-kappaB-DNA binding activity, expression of COX-2 protein, and neointima thickness and plaque area of aorta of the HC + A group were (5.14 +/- 0.32) microg/ml, (14.6 +/- 2.7) microg/ml, (0.342 +/- 0.02)A, (165 +/- 24) microm, and (24.3 +/- 7.6)% respectively, all significantly lower than those of the HC group [(9.39 +/- 0.79) microg/ml, (32.4 +/- 4.7) microg/ml, (0.572 +/- 0.061) A, (337 +/- 64) microm, and (49.5 +/- 21.3)%, all P < 0.05). By reducing the expression of COX-2. PMID- 18396631 TI - [Repair of facial nerve with bioactive artificial nerve conduit: experiment with rats]. AB - OBJECTIVE: To evaluate the effect of bioactive artificial nerve conduits in the repair of facial nerve defects. METHODS: Schwann cells (SC) were collected from the sciatic nerve and brachial plexus of a newborn SD rat, cultured, transfected with the plasmid pcDNA3. 1 (+)/GDNF containing glial cell line-derived neurotrophic factor gene, digested by pancreatin to make mixture of cell suspension to be injected into poly (L-lactic-co-glycolic acid) conduits. After injected with SCs, the conduits were cultured in for 2 weeks. Thirty female SD rat underwent buccal branch of facial nerve and then were randomly divided into 3 groups: Group A, undergoing direct anastomosis of the amputated dextral nerve; Group B, in which the 2 sides of amputated nerve were put into the PLGA conduit without transfected GDNF; and Group C in which the 2 sides of amputated nerve were put into the PLGA conduit with transfected GDNF. The motion of the facial muscles and whisker was observed. Two weeks, and 1, 2, and 3 months after the operation electrophysiological study was conducted to measure the action potential of the facial nerve. Then the rata were killed with their bilateral facial nerve and pontes were taken out to undergo histological examination. RESULTS: Compound action potential could be detected in all groups at any time points except in Groups A and B 2 weeks after the operation. The values of compound action potential of Group C at different time points were all significantly higher than those of Groups A and B (all P < 0.01). However, there were not significant differences in the compound action potential values at any time point between Groups A and B (all P > 0.05). The mean number of myelinated nerve fibers, thickness of myelin sheath, and number of motor neuron of Group C were all significantly higher than those of Groups A and B (all P < 0.05). CONCLUSION: Conduit with SCs transfected with GDNF gene + PLGA is superior to that with SCs + PLGA and direct anastomosis in repair of facial nerve. PMID- 18396632 TI - [Fluorescent imaging of angiogenesis and spontaneous metastasis in transplanted hepatoma: experiment with mice]. AB - OBJECTIVE: To establish stable enhanced green fluorescent protein (GFP) expressing human adenoid cystic carcinoma (ACC-M-GFP) cell lines with high lung metastatic potential, and to study the value and influential factor of in vivo monitoring of metastasis and angiogenesis by fluorescent imaging. METHODS: Human adenoid cystic carcinoma cell line ACC-M-GFP labeled with enhanced GFP was established. ACC-M-GFP cells of the dose 10(7)/ml were injected subcutaneously into two nude mice. Once the subcutaneous tumor reached 0.5 cm in diameter, it was removed, cut into 1 mm3 pieces, and implanted into the left hepatic lobe of 8 nude mice. The animals were observed by whole-body optical imaging system and stereofluorescent microscope each two weeks from the fourth week after transplantation. One mouse was killed and autopsied after each examination. The livers, lungs, and adjacent organs, such as diaphragm and lymph nodes were excised, and fluorescent imaging of different organs separately was studied. Hepatoma was cryostat sectioned and angiogenesis was visualized under fluorescence microscope. The findings of fluorescent imaging were compared with those of HE staining on routine paraffin sections. RESULTS: Liver tumors were transplanted successfully in all 8 nude mice. By whole-body optical system, the fluorescent signal from the liver tumor was detected through a skin-flap window after 6 - 8 weeks. Tumors were visualized directly through skin after 8 - 10 weeks. The hepatoma became enormous after 12 - 14 weeks, when the fluorescent signal of abdominal metastasis was detected. Metastasis of lung and lymph node was found by single organ imaging after 16 -18 weeks. By stereofluorescent microscope, microvessels were detected after 4 weeks, and a large quantity of tumor vessels like black branch stripes were found after 12 - 14 weeks. The tumor vessels appeared as dark tubiform network against the green fluorescence of the implanted tumor in section. CONCLUSION: Fluorescent imaging provides a method to monitor tumor growing, infiltrating, metastasis and angiogenesis in real time in vivo. PMID- 18396633 TI - [Progress in treatment of metastatic tumors in the bone by bisphosphonate]. PMID- 18396634 TI - [Antitumor immune responses induced by idiotype-pulsed dendritic cells with cell penetrating peptide vaccination in vivo]. AB - OBJECTIVE: To confirm the therapeutic effect of dendritic cell (DC) vaccine on treatment for mice with lymphoma and the protective effect of DC vaccine loaded with different antigens on the tumor-bearing BAL B/c mice. METHODS: Firstly, a mouse tumor model was set up by s. c. inoculation of 1 x 10(6)/mouse A20 tumor cells. Then different DC vaccines were injected, respectively, and the tumor size and survival time were observed. Secondly, the immunized mice with DC vaccines were challenged with A20 tumor cells, and observed whether a new tumor occurred in the mice and the time of survival. RESULTS: The tumor of mice immunized with Id-DC vaccines grew slower than the controls (mean time of survival was 40.4 days vs. 33.4 days), but statistically not significantly different. The tumor of mice injected with CPP-Id-DC vaccines grew slower than that injected with Id-DC vaccines and controls, and one of 5 mice got CR and the tumor in another one mouse became stable. The median survival time was 70.8 days during a 90-days observation period. The difference was significant (P<0.01). The mice injected with Id-DC vaccines were challenged with A20 tumor cells showed new tumor occurred at 7 - 12 days, and 1 of the 5 mice survived for 60 days. The mice injected with CPP-Id-DC vaccines had no tumor. CONCLUSION: The DC loaded with CPP Id was better than that loaded with Id alone in treating B cell lymphoma, and It can enhance their antitumor responses and prolong the survival time of the A20 tumor animal models. The vaccine of DC loaded with CPP-Id can protect mice from A20 tumor cell challenge. PMID- 18396635 TI - [The anti-tumor activity of GM-CSF-modified lung cancer cell vaccine and its synergism in combination with chemotherapy]. AB - OBJECTIVE: To investigate the anti-tumor effects and mechanism of tumor vaccines and whether chemotherapeutic agents administered prior to immunotherapy could augment the efficacy of the vaccines. METHODS: C57/BL mice inoculated with Lewis lung cancer cells were used as tumor models. Granulocyte-macrophage colony stimulating factor (GM-CSF) gene modified LA795 and Lewis lung cancer cell lines were administered as allogeneic and autologous tumor vaccines, respectively. After Lewis cells (1 x 10(7)) inoculation, the mice received irradiated GM-CSF secreting cancer vaccine solely or in combination with carboplatin. The survival of the mice was observed. The cytotoxicity of spleen cells or purified CD8(+) cells was analyzed by lactate dehydrogenase (LDH) assay. Serum level of IL-4 and IFN-gamma was detected using ELISA method. RESULTS: The cytotoxicity of the spleen cells or purified CD8(+) T cells against Lewis cells in the mice immunized with cancer cell vaccine was significantly increased, relative to that of the control, untreated group (P < 0.05). Serum level of Th1-type cytokine IFN-gamma was increased after vaccination, whereas Th2-type cytokine IL-4 showed no significant change. The GM-CSF secreting cancer cell vaccine had no significant influence on the survival of the mice with established heavy tumor burden. The combination of chemotherapy and cancer vaccine could statistically prolong the survival time; whereas any method itself had no significant effect. CONCLUSION: The GM-CSF secreting cancer cell vaccine can induce immune responses. The chemotherapeutic agents may be beneficial to enhance the anti-tumor activity of cancer vaccine. PMID- 18396636 TI - [Recombinant adenovirus-mediated overexpression of TIMP-1 effectively suppresses hepatocellular carcinoma cell line HepG2 in vitro and in vivo]. AB - OBJECTIVE: To explore the effects of overexpression of human tissue inhibitors of metalloproteinase-1 (hTIMP-1) on proliferation, invasion, metastasis, angiogenesis, and apoptosis in human hepatocellular carcinoma (HCC) cells in vitro and in vivo. METHODS: Recombinant adenoviral vector containing hTIMP-1 (AdhTIMP-1) was constructed previously. HepG2 cells were infected by AdhTIMP-1 and the changes of cell proliferation and invasion were detected in vitro. The anticancer activity of AdhTIMP-1 was evaluated in BAL B/c mice bearing HCC. Tumor volume and pulmonary metastases were observed. The mechanisms underlying the antitumor effect in vivo were investigated based on detection of microvessel density and apoptosis in tumor tissues. RESULTS: The resultant AdhTIMP-1 was successfully constructed and the expression of hTIMP-1 was detected by Western blot and RT-PCR. AdTIMP-1 could effectively infect HepG2 cells and significantly inhibit the proliferative activity and invasive ability of the tumor cells. Compared with the controls, pre-infection of HepG2 cells by AdhTIMP-1 resulted in a significant inhibition of tumor formation by 75. 8%. A single local injection of AdhTIMP-1 into pre-established tumors significantly reduced the tumor growth rate by 45.4%, tumor-associated angiogenesis index by 47.8%, lung metastases by 70.4%, and showed a 3-fold increase of apoptotic tumor cells. CONCLUSION: Our data indicated that AdhTIMP-1 can significantly attenuate tumor proliferation and invasion, reduce metastasis, inhibit angiogenesis, and induce apoptosis in HCC bearing mice and may pave the way for further liver cancer gene therapy. PMID- 18396637 TI - [Treatment of mouse liver metastasis by intraportal injection of Adv-p53]. AB - OBJECTIVE: To investigate the anti-tumor effect of intraportal administration of Adv-p53 in the treatment of the liver metastasis in mice. METHODS: 2 x 10(5) of MCA-205 cells were injected into the mouse portal vein to establish a murine liver metastasis model. The spleen was transpositioned subcutaneously to enable the administration of Adv-p53 continually into the portal system. Different doses of Adv-p53 were injected intraportally, while HBSS and Adv-CMV were injected intraportaly in the control group. Tumors in the liver were examined on day 21 after Adv-p53 administration. RESULTS: The liver weight in the Adv-p53 treated mice on day 0 group (1.20 +/- 0.34 g) was significantly less than that in the Adv CMV group (2.59 +/- 0.48 g, P < 0.05). The number of metastatic nodules in the Adv-p53 treated mice on day 0 group (9.0 +/- 9.9) was significantly less than that in the Adv-CMV group (57.1 +/- 11.3, P < 0.05), indicating that intraportal administration of Adv-p53 inhibited the formation of liver metastasis. This anti tumor effect was in a dose-dependent manner. After the liver metastasis was formed, Adv-p53 was administered intraportally. The liver weight in the Adv-p53 treated mice on day 5 group (1.22 +/- 0.09 g) was significantly less than that in the Adv-CMV group (3.98 +/- 1.01 g , P < 0.05). The number of metastatic nodules in the Adv-p53 treaed mice on day 5 group (5.5 +/- 3.5) was significantly less than that in the Adv-CMV group (113.2 +/- 5.8, P < 0.05). Repeatedly intraportal administration of Adv-p53 could enhance this anti-tumor effect. CONCLUSION: Local administration of Adv-p53 into the portal system would be a useful strategy for the liver metastasis treatment. PMID- 18396638 TI - [Apoptosis in esophageal cancer cells induced by all-trans retinoic acid]. AB - OBJECTIVE: To study the anti-tumor effects of all-trans retinoic acid (ATRA) and mechanisms of its action. METHODS: Human esophageal carcinoma cell line EC9706 cells were treated with ATRA at different concentration. The proliferation inhibition was examined by MTT assay. Morphological examination, TUNEL method and flow cytometry were used to detect the apoptosis and changes of cell cycle. Immunohistochemical method was used to detect the expression of apoptosis-related genes caspase-3 and bcl-2. The semi-quantification of protein expression was analyzed by pathological image analysis. RESULTS: ATRA inhibited the proliferation of EC9706 cells moderately. Apoptosis in EC9706 cells was induced by ATRA treatment. The morphology of EC9706 cells showed changes such as nuclear chromatin condensation and fragmentation. Sub-G1 peak was found by flow cytometry. The maximal apoptosis rate was 32.6%. The expression of caspase-3 gene was enhanced. The expression of bcl-2 gene was decreased. All these effects were presented in a dose-dependent and time-depend manner. CONCLUSION: Apoptosis is one of the key mechanisms of ATRA action on EC9706 cells. PMID- 18396639 TI - [Inhibitory effect of oxaliplatin in combination with hyperthermia on angiogenesis]. AB - OBJECTIVE: To study the effect of oxaliplatin in combination with hyperthermia on angiogenesis in vitro and in vivo. METHODS: MTT method was used to observe the influence of oxaliplatin on the proliferation of human umbilical vein endothelial cells (HUVEC) or human colon cancer cells (LOVO). The influence of oxaliplatin on HUVEC migration was evaluated by Transwell. Chick embryo chorioallantoic membrane (CAM) model was used to check whether the neovascularization of CAM could be suppressed in vivo. RESULTS: The survival rate of HUVEC was 80.1% - 42.5% within a range of 0.5 - 16 microg/ml and was negatively correlated with the concentration (correlation coefficient was - 0. 943, P = 0.005). The survival rate of LOVO cells within those doses was more than that of HUVEC. There was a synergistic antiangiogenic effect when a combination of oxaliplatin (0.5 microg/ml, 1 microg/ml and 16 microg/ml) with hyperthermia was used while additional effect was shown by the combinatioin of oxaliplatin (2 microg/ml, 4 microg/ml and 8 microg/ml) and hyperthermia in vitro. Oxaliplatin inhibited migration of HUVEC in vitro at low doses (0.25 - 2 microg/ml), and also suppressed angiogenesis of CAM in vivo at doses of 1 -4 microg/ml. CONCLUSION: The results of this experiment showed that low dose of oxaliplatin has anti angiogenic effect in vitro, while in combination with hyperthermia has additional effect both in vivo and in vitro. PMID- 18396640 TI - [The expression of human macrophage metalloelastase mRNA in gastric cancer cell lines and tissues and its clinical significance]. AB - OBJECTIVE: To make measurement of the expression of HME mRNA in both gastric cancer cell lines and tissues and evaluate its role in development of gastric cancer. METHODS: The HME mRNA expression in 3 gastric cancer cell lines and tissues was detected by real-time fluorescent quantitative PCR. RESULTS: All three gastric cancer cell lines expressed HME mRNA, and the HME mRNA expression level in gastric cancer tissues was higher than that in normal tissues (P <0.05). The rate of lymph node metastasis of HME mRNA positive cases was lower than that of HME mRNA negative ones (P < 0.05). Expression of drug resistance gene GST of HME mRNA positive cases was lower than that of HME mRNA negative ones (P <0.05). The two-year survival rate of HME mRNA positive cases was higher than that of HME mRNA negative ones (P < 0.05). There was no correlation between the expression of HME gene and the tumor location, size, depth of invasion, degree of malignancy, expression of drug resistance gene top II and PG. CONCLUSION: The expression of HME gene in gastric cancers may be related with lower possibility of metastasis and predict a better prognosis. PMID- 18396641 TI - [Expression and clinical significance of CD44v6 and sCD44v6 in gastric carcinoma]. AB - OBJECTIVE: To evaluate the correlation of CD44v6 and sCD44v6 expression with the biological activity of gastric carcinoma. METHODS: Mucosa samples from 103 gastric carcinoma patients and 10 healthy persons (control) were examined using immunohistochemical SP method; sCD44v6 level in peripheral blood samples was detected with ELISA in 86 gastric cancer patients, 30 gastric ulcer patients and 30 healthy controls. The follow-up period was 3-91 months for 88 patients in CD44v6 group and 1-91 months for 55 patients in sCD44v6 group. RESULTS: (1) Positive expression of CD44v6 was found in 60.2% of gastric carcinoma, where as 0% in the normal gastric mucosa. CD44v6 protein expression was positively correlated with TNM stage, perigastric lymph node metastasis, presence of cancerous embolic angiopathy and Borrmann classification (P <0.05), but was not correlated with invasion depth, differentiation, metastasis and survival (P > 0.05); 77.8% of the patient with liver metastasis had a strong CD44v6 protein expression. (2) sCD44v6 level in the peripheral blood of gastric carcinoma patient was significantly higher than that in gastric ulcer patient or healthy control; Compared with palliative gastrectomy, sCD44v6 level significantly went down after radical operation. No relationship between sCD44v6 level and the pathological features was found, which included invasion depth, perigastric lymph node metastasis, presence of cancerous embolic angiopathy, differentiation and Borrmann classification; The survival in the high sCD44v6 level group was longer than that in low sCD44v6 level group ( P = 0.0281), but no significant difference was observed by Cox Regression analysis (P = 0.415). (3) No apparent correlation was observed between CD44v6 expression in gastric cancer and sCD44v6 concentration in peripheral blood (P>0.05). CONCLUSION: Detection of CD44v6 expression in the gastric cancer may be helpful in evaluating the biological features and the survival in gastric carcinoma. The level of sCD44v6 in the blood may be also helpful in differential diagnosis, evaluation of surgical treatment and biological activity for gastric cancer. No apparent correlation is observed between CD44v6 expression in the cancer and that of sCD44v6 in the blood. PMID- 18396642 TI - [Anti-tumor effects of chemotherapeutic drugs on human gastric cancer cells in vitro and the relationship with expression of hTERT mRNA]. AB - OBJECTIVE: To evaluate in vitro antitumor effects of chemotherapeutic drugs, and investgate the relationship with expression of hTERT mRNA in human gastric cancer tissues. METHODS: Fresh samples of gastric cancer obtained from operation room were prepared to single-cell suspension (3 x 10(5) to 5 x 10(5) cells ml(-1)) and were separately exposed to taxol (TAX), cisplatin (CDDP), 5-fluorouracil (5-Fu), adriamycin (ADM), mitomycin (MMC) for 48 hours. Metabolic activity and inhibitory rate of the cells were determined by trypan blue exclusion and MTT assay. Expression of hTERT mRNA was detected by in situ hybridization (ISH). RESULTS: The inhibition rate of cancer cells exposed to chemotherapeutic drugs was different, and that of TAX, CDDP, 5-Fu was significantly higher than that of ADM and MMC. The positive rate of hTERT mRNA expression was 90.0% (54/60) and positive cells showed resistance to 5-Fu and ADM. CONCLUSION: Overexpression of hTERT mRNA may contribute to primary drug-resistance of tumors. Chemosensitivity tests by MTT assay may contribute to prediction of effectness in applying chemotherapeutic drugs and identify drug-resistant cases. PMID- 18396643 TI - [Hypermethylation and regulation of expression of secreted frizzled-related protein genes in colorectal tumor]. AB - OBJECTIVE: To investigate the functions of promoter hypermethylation of secreted frizzled-related protein (sFRP) genes in colorectal tumorigenesis and progression. METHODS: Three colorectal cancer cell lines, RKO, HCTll6 and SW480, were treated hy 5-aza-2'-deoxycytidine and trichostatin A for demethylation. The promoter hypermethylation and expression of sFRP genes in colorectal tumor tissue and colorectal cancer cell lines were detected hy methylation-specific PCR and reverse transcription PCR, respectively. RESULTS: None of the normal colorectal mucosa tissues showed methylation of sFRP genes. sFRP1, 2, 4 and 5 were frequently methylated in colorectal adenocarcinoma, adenoma and aberrant crypt foci (ACF) (sFRP1 > 85%, sFRP2 > 75%, sFRP5 > 50%), the differences between any two of them were not significant (P >0.05). Methylation was more frequent in colorectal tumors than in normal mucosa and adjacent normal mucosa from patients with tumor. Hypermethylation of sFRP genes was present in three colorectal cancer cell lines. When sFRP genes were methylated, their corresponding mRNA expression was absent. After cells were treated by DAC/TSA combination, the silenced sFRP expression could be effectively re-expressed. CONCLUSION: Hypermethylation of sFRP genes is a common early event in the evolution of colorectal tumors that occurs frequently in ACF. Methylation of sFRP1, 2 and 5 genes might serve as biomarkers for the early detection of colorectal tumors. Demethylation can effectively reverse gene expression that appears possibly to be an effective way for tumor therapy. PMID- 18396644 TI - [MRI in the evaluation of peripancreatic vessel invasion and resectability of pancreatic carcinoma]. AB - OBJECTIVE: To investigate the role of MRI in evaluating the peripancreatic vessel invasion and resectability of pancreatic carcinoma based on the comparison of MRI image with surgical exploration, and try to establish the criteria for assessment of the sensitivity, specificity and accuracy of resectability. METHODS: Forty-one pancreatic carcinoma patients confirmed by pathology received preoperative plain and contrast enhanced MRI scan, and 37 of them had additional coronal MRA scan. Peripancreatic vessel invasion was preoperatively assessed based on MRI features, and the vessel invasion degree from the uninvolved to the severely involved was divided into 6 grades represented by 1, 2a, 2b, 3a, 3b and 4, respectively. Compared with the findings during the surgery, the sensitivity and specificity of each vessel invasion grade were studied and the receiver operator characteristic curve (ROC) was drawn. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of resectability evaluation based on 2 and 2a degree were calculated respectively. The resectability of involved arteries and veins of grade 2 were also analyzed. RESULTS: Of the 41 patients, 22 had resectable tumor, with 20 curative resection and 2 palliative. Compared with the findings during surgery, seven vessels including three arteries and four veins were not correctly interpreted by MRI. If grade 1,2a,2b,3a and 3b was used as the resectable standard,respectively, the sensitivity to predict the unresectbility was 78.3%, 84.8%, 67.4%, 56.5% and 47.8%, respectively. Receiver operator characteristic curve demonstrated that grade 2a was the optimal critical point. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of grade 2a in predicting the unresectbility were 84.8%, 98.5%, 92.9%, 96.6% and 95.9%. CONCLUSION: Our data showed that grade 2a (tumor involvement < 2 cm long and < 1/2 circumference of the vessel) may be more sensitive and accurate in predicting the resectability, which may be considered as the line of demarcation between the respectable and unresectable cases in clinical practice. PMID- 18396645 TI - [Expression of cytokines Th1 and Th2 in patients with esophageal squamous cell carcinoma]. PMID- 18396646 TI - [Clinical value of multidetector computed tomography in detecting lymph node metastasis of early gastric cancer]. AB - OBJECTIVE: To evaluate multidetector computed tomography (MDCT) as a pre operative staging tool for detecting lymph node metastasis in patients with early gastric cancer (EGC). METHODS: In 77 consecutive patients with EGC, lymph node metastasis was assessed pre-operatively with MDCT at a slice thickness of 2.5 - 5.0 mm (n=24) or 7.5 - 10.0 mm (n =53). RESULTS: Overall accuracy of detecting NO to N3 lymph node was 74.0% for MDCT and 54.5% for operative assessment. The sensitivity, specificity, positive predictive value and negative predictive value in detecting lymph node metastasis was 75. 0%, 65.0%, 30.0% and 92.9% with 2.5 - 5.0 mm MDCT image, which was 62.5%, 82.2%, 38.5% and 92.5% by 7.5 - 10.0 mm MDCT image, whereas it was only 45.5%, 63.6%, 17.2% and 87.2% by operative assessment. CONCLUSION: Pre-operative assessment by multidetector CT may have a high accuracy in detecting lymph node metastasis for patients with early gastric cancer. PMID- 18396647 TI - [Value of multidetector computed tomography in differentiating ovarian tumors from non-ovarian masses]. AB - OBJECTIVE: To evaluate the ability of multidetector computed tomography (MDCT) in differentiating ovarian tumors from non-ovarian masses. METHODS: Forty-two cases with pelvic masses were examined with 16-row MDCT. All source image of each case was put into workstation for multi-planar reconstruction (MPR) and curved planar reconstruction(CPR). Axial image combined with 2D image was used for determining the relationship of the mass to ovarian vascular pedicle and identifying the normal ovary, which was compared with postoperative pathologic result and the finding during operation. All the data was compared using Fisher's exact test. RESULTS: There were 28 ovarian tumors and 14 non-ovarian tumors in this series. If the ovarian vascular pedicle sign was used for determining whether the tumor was from the ovary or not, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy was 89.3%, 85.7%, 92.6%, 80.0% and 88.1%, respectively, with a significant difference in differentiating the tumor from the ovary or non-ovarian organs (P <0.05). If the identification of full normal ovary was used to determine non-ovarian origin of the tumor, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy was 50.0%, 100.0%, 100.0%, 80.0% and 83.3%, respectively, also with a significant difference in differentiating the tumors from the ovary or non ovarian organs (P <0.05). CONCLUSION: MDCT can clearly show the relationship of the tumor to the normal ovary and its vascular pedicle, which is very helpful in differentiating the ovarian tumors from a non-ovarian masses. PMID- 18396648 TI - [CT image features of 46 cases with pneumonic-type lung cancers]. AB - OBJECTIVE: To analyze the CT image features of pneumonic-type lung cancer and to reduce misdiagnosis. METHODS: The CT findings of 46 patients with pneumonic-type lung cancer were retrospectively reviewed, and CT image in the differential diagnosis of this special kind of disease was evaluated. RESULTS: According to the extent of lesion, these cases were divided into two groups: multi-lobe consolidation group and single lobe consolidation group. The lesions in the latter group located in the upper, middle or lower lobe, respectively. Twenty nine cases had homogeneous consolidation lesion, 14 cases showed single or multiple cysts and cavities in the lesions, 3 cases exhibited localized low density in the lesion. Forty-one cases shown the sign of air bronchogram with presentation of narrow air bronchogram in 25 of those. Forty cases showed well or ill defined ground-glass opacitiy surrounding the consolidation lesion. Fifteen cases had multi-nodules or opacities distributed in centrilobular or centric bronchiolar location. Of the 30 patients who received contrast medium, 23 showed distinct enhancement, and 7 showed indistinct enhancement with a positive CT angiogram. CONCLUSION: CT findings including lower lobe distribution, homogeneous consolidation, narrow air bronchogram, well defined ground-glass and CT angiogram are helpful in differentiating pneumonic-type lung cancer from various kinds of infection. However, most of CT manifestations of pneumonic-type lung cancer are not specific. Therefore, it's necessary to combine CT findings with other clinical data when making diagnosis. PMID- 18396649 TI - [Surgial treatment and prognosis of ovarian metastasis from colorectal cancer]. AB - OBJECTIVE: To summarize the surgical treatment experiece and to investigate the prognosis of the patients with ovarian metastasis from colorectal cancer. METHODS: The data of 67 patients with synchronous or asynchronous ovarian metastasis from colorectal cancer surgically treated between January 1989 and December 2005 were collected and analyzed retrospectively using Statistical Package for the Social Sciences (Release 11.5, SPSS, Inc). Prognostic factors were analyzed using chi2 test. Survival analysis was estimated by the Kaplan Meier method and compared using the log-rank test. Multivariate analysis was carried out by Cox regression. RESULTS: The overall 1-, 3- and 5-year survival of these 67 patients was 71.0%, 18.7% and 9.2%, respectively. Univariate analysis revealed that the metastasis was confined in the ovary or pelvis only, unilateral/double ovarian metastasis, and operation mode were all statistically significant prognostic factors (P <0.05). Cox regression analysis showed that the operation mode was the most important prognostic factor (OR = 3.531, P <0.001). CONCLUSION: Surgical treatment is still the most effective mode in the treatment for the ovary metastasis from colorectal cancer. PMID- 18396650 TI - [Evaluation of efficacy and influence factors of transarterial interventional therapy in patients with liver metastasis from malignancy of alimentary tract]. AB - OBJECTIVE: To evaluate the efficacy and prognostic factors of transarterial interventional therapy (TAIT) in patient with liver metastasis from malignancy of the alimentary tract. METHODS: 266 patients with unresectable liver metastases from malignancy of the alimentary tract received totally 754 sessions of transarterial interventional therapy. Cox regression was used in the proportional hazard analysis. RESULTS: The overall response rate of TAIT was 45.4%, The median survival time (MS) was 14.3 months in this series. The 0.5-, 1-, 2-, 3-, 5-year cumulative survival rate (CSR) was 83.1%, 56.8%, 17.7%, 9.3% and 1.5% , respectively. No severe adverse reaction was observed except nausea, vomiting and mild fever as well as pain in the hepatic area. It was found that portal vein tumor thrombosis (PVTT), the blood supply of tumor, metastasis from esophageal carcinoma, the number of metastasis, multi-lobe involvement, resection nature of primary tumor were independent factors affecting survival. CONCLUSION: Transarterial interventional therapy is effective for treatment of liver metastasis from malignancy of the alimentary tract. Portal vein tumor thrombosis, metastasis from esophageal carcinoma, multiple metastatic lesions, multi-lobe involvement are poor prognostic factors, while complete resection of the primary tumor and rich blood supply of metastatic lesion are good independent prognostic factors. PMID- 18396651 TI - [Comparison of tolerance and toxicity of CEF-100 regimen versus CEF-60 regimen as adjuvant therapy for breast cancer]. AB - OBJECTIVE: To evaluate tolerance and toxicity of high-dose epirubicin regimen CEF 100 as adjuvant therapy for breast cancer. METHODS: From March 2005 to October 2006, 98 patients with stage I - III a breast cancer were randomly assigned to receive postoperative chemotherapy with CEF-100 regimen (epirubicin 100 mg/m2, dl per 21 days for 6 cycles, n =48) or CEF-60 regimen (epirubicin 60 mg/m2, dl per 21 days for 6 cycles, n = 50). Blood routine test were done every cycle, liver and kindey function were examined and adverse effects were recorded after every cycle. RESULTS: No difference of average leucocyte or neutrophil count (P >0.05) was observed in every cycle. Adverse effects of digestive tract and damage of liver function in CEF-100 group were more severe than that in CEF-60 group (P <0.05), but all adverse effects could be relieved by treatment. No severe non hematological toxicity and cardiac toxicity in both groups were observed (P <0.05). There was no death caused by chemotherapy. CONCLUSION: Our data shows that high dose epirubicin-containing CEF regimen is safe and tolerable for postoperative chemotherapy of breast cancer patient, and the adverse effects could be relieved by marrow support and liver-protection therapy. Further observation and longer follow-up is still needed in order to evaluate the efficacy of this high dose regimen. PMID- 18396652 TI - [Consensus on the medical treatment of gastrointestinal stromal tumors]. PMID- 18396653 TI - [Prevalence of preterm birth among singletons in 10 counties (cities) of China, 1993-2005]. AB - OBJECTIVE: To describe the secular trends and epidemiological characteristics of preterm birth among singletons in 10 counties (cities) of China during 1993-2005. METHODS: We analyzed data on 542 923 women (gestational age > or =28 weeks) collected through Perinatal Healthcare Surveillance System established by the Institute of Reproductive and Child Health, Peking University. Chi-square tests were employed to test the differences in prevalence of preterm birth among different groups. RESULTS: 25 784 preterm births were identified, including 1530 stillbirths. Preterm birth rate was 4.75% (95% CI:4.69-4.81) for all births and 4.49% (95% CI:4.44-4.55) for live births. Preterm birth rate declined steadily from 1993 to 2005 and had no significant seasonal variation. Preterm birth rate from the urban areas of the Southern part of the country,was higher than that in the rural areas which was also higher than that seen in the rural areas from the northern part of the country. Relations between women's age at delivery and preterm birth appeared to be U-shaped. Increased preterm birth rates were also observed in women with lower education level,more parities,and previous history of preterm birth or abortion. CONCLUSION: Preterm birth rate decreased steadily from 1993-2005 in 10 counties (cities) but varied by areas of the country. PMID- 18396654 TI - [Study on the distribution and risk factors of hypertension among children: a cohort study]. AB - OBJECTIVE: To explore the association between hypertension and the tendency of change among children,so as to lay a foundation for the prevention and control of hypertension. METHODS: Based on findings from the prevalence survey that carried out in September 1999 in Daqing of Heilongjiang province. New admission children were selected as subjects to conduct a five-year cohort study. All the subjects were interviewed with questionnaires and their blood specimens were collected for biochemical analysis. All data were analyzed using SPSS 10.0 software. Results The prevalence of hypertension among 447 children was found 2.01% at the baseline study but increased to 5.37% in the fifth year. During a five year period, the systolic pressure level among children increased from (100.65 +/- 11.62)mmHg (1 mm Hg = 0.133 kPa) to (106.67 +/- 9.29) mm Hg,while the diastolic pressure level was from (66.27 +/- 11.31) mm Hg to (70.28 +/- 7.98) mm Hg and showed significant difference between boys and girls. There were association between hypertension and family history, body mass index (BMI), triglyceride, insulin, insulin resistance index while insulin sensitivity index and family history, BMI and insulin sensitivity index appeared to be the important factors. Children under this study were divided to 'with family history or without' and then every group was divided to 'with over weight-obesity or normal'. Obesity and insulin sensitivity seemed the key risk factors on hypertension. Descent of insulin sensitivity was an independent risk factor. CONCLUSION: The level of blood tension among children in Daqing city was higher than that from the national data. The present study confirmed that over-weight,obesity, heredity and insulin resistance were the risk factors of hypertension while insulin resistance was related to hypertension. The interaction of these risk factors was independent or correlated to each other. PMID- 18396655 TI - [A cohort study on parental history and risk factors of stroke in Chinese population]. AB - OBJECTIVE: To assess the relationship between parental history and the incidence of stroke in Chinese populations. METHODS: A total of 15,131 individuals (7015 men and 8116 women, aged from 35 to 59 years) in China were enrolled and followed up from 1992 to 2005. Information on the parental history of stroke of the participants was collected at baseline study under face to face interview. Individuals were divided into three groups according to their parental history of hypertension: those with no parental history of hypertension (Group 1), those with only one parent having the history of hypertension (Group 2) and those with both parents having the history of hypertension (Group 3). The hazard ratio and its 95% confidence interval were calculated using the Cox proportional hazard model. RESULTS: There were 370 stroke events during 163 858 person-years of observation. After adjusting for age and sex, the hazard ratioon stroke for group 1, group 2 and group 3 were 1.00, 1.74 (1.33-2.29), 3.61 (1.86-7.01), respectively. After adjusting for age, sex, smoking, drinking, serum total cholesterol (TC), high density lipid cholesterol (HDL-C), fasting glucose, body mass index (BMI) and systolic blood pressure (SBP) in the model, the hazard ratio for stroke of those three groups were 1.00, 1.34 (1.02-1.77), 2.50 (1.29-4.87), respectively. CONCLUSION: Individuals with parental history of stroke had a higher risk of stroke, and this was especially true for those with both parents having the history of stroke. PMID- 18396656 TI - [Study on C-reactive protein and the risk factors of traditional coronary heart disease]. AB - OBJECTIVE: To ascertain the relationship between C-reactive protein (hs-CRP) and risk factors of traditional coronary heart disease (CHD) in a sampled population from Jiangsu province of China. METHODS: Using data derived from a project on multiple metabolic disorders and metabolic syndrome in Jiangsu province, a cross sectional study was carried out. Criteria was set as: CRP level categories from 1 to 2.99 mg/L, and greater than 3 mg/L, to explore the relationship between different category of CRP and risk factors of traditional coronary heart disease which would include body mass index (BMI), cigarette smoking, high blood pressure, blood glucose, total cholesterol level, triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) level, physical activity etc. RESULTS: The attack rate of high CRP increased with the degree of risk factors related to traditional coronary heart disease. However, when hs-CRP fell between 1-2.99 mg/L, part of the traditional risk factors would display significance. Significance increased in high CRP related to grading was observed across the Framingham heart score (quintiles). Data from the logistic regression analysis showed that: when hs-CRP (> or =3 mg/L), the OR of BMI, TG, high blood pressure, HDL-C, cigarett smoking, blood glucose all appeared statistical significance whereas hs-CRP between 1-2.99 mg/L, the OR of trational CHD risk factors did not show statistical significance but 60.98% of the males and 59.02% of the females would have high hs-CRP incriminate traditional CHD risk factors. CONCLUSION: As the close relationship between traditional CHD risk factors and hs-CRP was seen, the role of hs-CRP in estimating the risk of CHD was limited, despite the origin of data, either from clinical or from cohort study. PMID- 18396657 TI - [Study on the impact of related psychological stress and fatigue to different smoking behaviors among undergraduate students]. AB - OBJECTIVE: To explore the relationship between psychological stress, fatigue and different smoking behaviors among undergraduate students. METHODS: 8138 undergraduate students in grade 1 to 3 were sampled with cluster sampling method from a comprehensive university in Guangzhou, and were studied on their psychological stress, peer and self smoking behaviors, as well as socio demographic factors with a self-administered questionnaire. Physical and mental fatigue was measured by Chalder fatigue scale. A multinomial logistic regression analysis was performed to explore the association between psychological stress, fatigue and different smoking behaviors with 'non-smokers' defined as control group while 'triers' and 'current smokers' as case group, respectively. RESULTS: There were students trying to smoke or becoming current smokers on university campus. After adjusting the effect of relevant socio-demographic factors and peer smoking behavior, physical fatigue (OR = 1.044, 95% CI: 1.013-1.075) and 'worry about current academic and future work' (OR = 1.020, 95% CI: 1.001-1.040) were the risk factors of trying to smoke among male students, while 'study stress (OR = 1.063,95% CI: 1.012-1.117) was the risk factor for female students when start smoking. 'Personnel relationship and environment stress' (OR = 1.152, 95% CI: 1.012-1.312) were the risk factors for female students' current smoking behavior. CONCLUSION: Psychological stress and fatigue were moderately associated with smoking behavior among undergraduate students and the impact was different on different gender and smoking behaviors that suggesting that some specific psychological measures should be taken so as to control different smoking behaviors among undergraduate students. PMID- 18396658 TI - [Effects of traffic exhausts on children' s neurobehavioral functions scored by neurobehavioral evaluation methods in Quanzhou, Fujian province]. AB - OBJECTIVE: To explore the effects of traffic exhausts on children's neurobehavioral functions. METHODS: A field study was conducted in Quanzhou, Fujian province where two primary schools were chosen based on the numbers of automobiles passing by,and the data of ambient air pollutants from the monitoring system. School B and School A located at the high traffic exhausts pollution area and the clear area, respectively. Neurobehavioral functions of pupils in grade 2 and 3 were scored. RESULTS: School B had very similar score of Ruiwen Test with School A, while the score of Digit Eras Test was lower than that in School A with marginal significance. The scores of Digit Symbol,Aim Tracing, Sign Register and Cormect Number test were 38.5 +/- 9.96, 84.3 +/- 27.83, 37.4 +/- 11.62 and 17.1 +/- 5.88 in School B,respectively, which were significantly lower than those in School A (41.6 +/- 12.97, 95.5 +/- 35.80, 42.3 +/- 15.58 and 18.7 +/- 5.78) respectively. Scores in Digit Symbol,Aim Tracing,Sign Register and Cormect Number of pupils in School B were 2.9, 11.5, 5.6 and 1.6 lower than those in School A after adjusting on other confounding factors. CONCLUSION: It was suggested that traffic exhausts might cause damage to children's neurobehavioral functions. PMID- 18396659 TI - [Application of BED-CEIA to estimate the human immunodeficiency virus prevalence among injecting drug users in Chongqing municipality]. AB - OBJECTIVE: To estimate the human immunodeficiency virus (HIV-1) prevalence of injecting drug users (IDUs) in Chongqing city. METHODS: To apply BED-capture enzyme immunoassay (CEIA) which was based on the principle of HIV-antibody varies as the disease progress, in order to estimate both the HIV incidence and prevalence of IDUs from two IDUs surveillance sites in Chongqing. RESULTS: During the research period, 4711 serum samples were tested by ELISA and 130 were HIV-1 positive, confirmed by Western blot. The prevalence of IDUs surveillance site A from 1999 to 2006 were 0.73%, 2.02%, 1.54%, 2.96% and 2.80%, and the incidence rates were 0.57%, 0.93%, 0,1.24% and 1.68% respectively. The prevalence of IDUs surveillance site B appeared to be 4.21%, 9.96%, 8.13%, and the incidence rates were 0.95%, 1.04% and 0.90% respectively, from 2004 to 2006. CONCLUSION: Many of the IDUs HIV carriers in Chongqing had been infected for long time, and the incidence rates among them were steady, keeping at the same level for 1-2 years. Promotion on intervention for IDUs had produced certain effects but more attention still needs to be paid. PMID- 18396660 TI - [The development and assessment on an instrument regarding quality of life for patients living with HIV/AIDS in China]. AB - OBJECTIVE: To develop a specific quality of life (QOL) instrument for Chinese patients living with HIV/AIDS and to assess its reliability and validity. METHODS: A primary questionnaire was developed with reference to existing instrument and information through discussion with experts, front-line professionals and patients living with HIV/AIDS. This questionnaire was applied in 443 patients living with HIV/AIDS. Some items were removed through the results of responsive rate, factor analysis, correlation matrix,internal consistency and so on. Reliability and validity were assessed after items reduction. RESULTS: The developed instrument contained 44 items, including 10 scales and other 3 items used for the evaluation of curative effect and side effect of anti-virus drug. No substantial ceiling/floor effects existed. All scales' internal consistency coefficients were > or = 0.70 and the whole instrument was 0.90. The two week retest reliability coefficient of the whole instrument was 0.80, and there were 6 scales below 0.70 of this coefficient. Multitrait/multiitem assessment indicated scaling success rates were all 100% for all items. Spearman correlation coefficient of total score of this instrument with SF-36 was 0.69. CONCLUSION: It was the first time that the developed QOL-CPLWHA instrument introducing the scale of hostility regarding mentality trend as well as expressed its acceptable reliability and validity so could be used for the evaluation of Chinese patients living with HIV/AIDS. PMID- 18396661 TI - [Study on the relations between psycho-behaviors and weight control in adolescents]. AB - OBJECTIVE: To describe the different sex, age and grade distribution of coping behavior and its psychological effects on weight, and to analyze the correlation between overweight, obesity and unhealthy psycho-behaviors related to weight control in adolescents. METHODS: A survey was conducted among students of grade 1 and grade 2 at a junior middle school in Hefei. Students completed the anonymous. questionnaire including the demographic characteristics of adolescents and parents, self-assessed weight status and adolescents' psycho-behaviors related to weight control. Anthropometric measures were taken together with body mass index (BMI) calculated. Students were divided into obesity,overweight and control groups by BMI separate criteria and by self reporting, respectively. RESULTS: Among all the students, 17.4% reported having unhealthy psychological feelings related to weight control including 3.8% reported having unhealthy behaviors related to weight control and 19.1% of them reported having unhealthy psycho behaviors related to weight control. The rates of unhealthy psychological feelings, unhealthy behaviors and total unhealthy psycho-behaviors related to weight control among girls (20.6%, 4.3%, 22.6%) were all higher than those among boys (14.5%, 3.3%, 15.8%) (P < 0.05). The rates of unhealthy psychological feelings, unhealthy behaviors and total unhealthy psycho-behaviors related to weight control in obese individuals both assessed by BMI and assessed by self reporting appeared to be the highest (P < 0.05). In addition,data showed that obesity and overweight assessed both by BMI separate criteria and by self reporting appeared positive correlations with the rates of unhealthy psycho behaviors related to weight control. Moreover, the relationship between obesity or overweight assessed by the two methods and unhealthy psycho-behaviors related to weight control did not alter by other factors. CONCLUSION: Unhealthy psycho behaviors related to weight control of girls are more severe than boys. The relationship between obesity and unhealthy psychological feelings and behaviors related to weight control does not alter by other factors such as social factors. PMID- 18396662 TI - [Analysis on the feasibility of reducing the concentration in edible iodine-salt based on the results of iodized salt monitoring program from the year of 2004 to 2006, in China]. AB - OBJECTIVE: To analyze the feasibility of reducing the concentration of iodized salt based on the results of iodized salt monitoring from the year of 2004 to 2006. METHODS: Special software for iodized salt monitor and SAS 9.0 were used to analyze salt monitoring data and urine iodine data of women at reproductive age in high-risk areas in 2006. RESULTS: Based on the data from monitoring program, adequate iodized salt coverage increased constantly in China. The quality of iodized salt was stable with less than 2 mg/kg iodine loss at production level but most was at 3 mg/kg iodine loss under estimation during the process of distribution from factory to households. Individual daily intake of iodized salt was higher than the recommendation from WHO but the average level of urinary iodine excretion of women and school children was more than adequate. CONCLUSION: To decrease the concentration of iodine in edible salt was necessary in China. Our findings provided recommendation on the concentration of iodine in edible salt that should be adjusted from the current concentration of 35 mg/kg to 25-28 mg/kg, and the variation should be controlled from the current range of +/- 15 mg/kg to +/- 10 mg/kg. PMID- 18396663 TI - [Genetic analysis on HA1 and NA regions of influenza virus subtype A3 isolates of Zhejiang province during 1998-2005]. AB - OBJECTIVE: To study the relationship between influenza epidemic and genetic characteristic on HA and NA regions of influenza virus subtype A3 isolates of Zhejiang province in the recent years. METHODS: RNA of 25 influenza virus subtype A3 isolates, circulated in Zhejiang province during 1998 to 2005, was extracted. HA1 and NA regions were amplified and sequenced. All the sequence data were analyzed using BioEdit. RESULTS: HA1 and NA regions of all the isolates belonged to 987nt and 1362nt, encoding protein of 329 and 454 amino acids respectively. Isolates shared amino acid homology of 90.9%-99.3% and 95.2%-99.5% on HA1 and NA regions, while divergence on HA1 was greater than that on NA region. During a period of 8 years, 30 amino acids on HA1 region were substituted and 14 of which refer to 4 antigenic determinant sites. Meanwhile,21 amino acids on NA region were substituted and 5 of which referred to 3 antigenic determinant sites. Significant divergences, both in HA1 and NA, were observed among isolates in 1998 and 2002, showing that they belonged to absolutely different branches. Additionally, influenza virus subtype A3 isolates identified in recent years, with 11 N-linked glyeosylation sites in HA1 region, had 5 sites more than early A/Aichi/2/68 strain. Since 1998,3 sites had been inserted in epidemic strains, indicating the accelerated trend of glyeosylation sites were increasing. CONCLUSION: There is a correlation between antigenic drift of influenza virus subtype A3 and the two epidemics in Zhejiang province in 1998 and 2002. PMID- 18396665 TI - [Study on the epidemiological characteristics of hemorrhagic fever with renal syndrome in Inner Mongolia]. AB - OBJECTIVE: In order to better understand the epidemiological features of Hantviruses in Inner Mongolia. METHODS: Epidemiological surveillance data during the period of the past 52 years were analyzed. An epidemiological survey was carried out in the main epidemic areas in 2005. RESULTS: A total of 8310 hemorrhagic fever with renal syndrome (HFRS) cases were reported in Inner Mongolia from 1955 to 2006, and distributed in 61 counties. HFRS cases were mainly distributed in the east part of Inner Mongolia before 1990. However, HFRS cases had occurred in the middle and western parts since 1990. Hulunbeier prefecture, from the eastern part of Inner Mongolia, had been the most severe area being hit by HERS since the first outbreak in 1955, with 7369 cases reported over the past 52 years, and accounted for 88.68% of the total cases in the whole autonomous region. Although no HFRS cases had been reported before 1999 in Bayannaoer which located in the western part of Inner Mongolia, a total of 95 cases were reported in 2005. Hantavirus antigens had been detected in 11 species of rodents so far,including Apodemus agrarius, Rattus norvegicus, Mus Musculus, Cricetulus barabensis, meriones meridianus, Microtus maximowiczii , Clethrionomys rutilus, Apodemus peninsulae, Phodopus roborvskii, Dipus sagitta and Allactaga sibirica. CONCLUSION: Results suggested that the epidemics might remain at a relatively high level in the years to come in Inner Mongolia. Furthermore, there might be other types of Hantaviruses in addition to the already identified Seoul viral type in this area. PMID- 18396664 TI - [Study on a high coverage of measles vaccine while high incidence of measles disease still appeared in Shaanxi province]. AB - OBJECTIVE: To analyze the affecting factors on the cause of measles and measles vaccine under the high coverage of measles vaccine in Shaanxi province. METHODS: Age distribution and vaccination history on measles cases were studied. Throat swabs were obtained from measles cases. Measles virus was isolated from collected specimens with phenol-chloroform extraction method. Amplification was performed by RT-PCR in order to amplify 450 bp fragment of the -COOH side of N gene,and then the sequences of PCR products were detected to confirm the gene type of measles virus. Sera were obtained from patients who were in acute phase of measles disease,and antibody titer against measles vaccine strain and wild strain were determined by small amounts neutralization test. RESULTS: Measles cases with the history of measles vaccination were accounted for 38.97% of the total numbers. The geometrical mean titer (GMT) (56.18) against S191 attenuated strain was significant higher than that of wild strain (26.90) among these measles patients with history of having received measles vaccination. The GMT (25.40) against S191 attenuated strain was similar to that of wild strain (27.86) among these measles patients with non-history of measles vaccination. The antibody negative rate against wild strain was 19.15% to these sera from patients with the history of measles vaccination and antibody potency against S191 strain was less than 16. CONCLUSION: The appearance of higher measles incidence under the higher coverage of measles vaccine indicated that measles epidemic strain might degenerate as the result regarding the failure of the present measles vaccine in protecting the transmission of H1 wild strain. PMID- 18396666 TI - [Evaluation on current malaria prevalence using capture-recapture method in national sentinel surveillance points malaria]. AB - OBJECTIVE: To explore the feasibility of using the capture-recapture method (CRM) for national malaria sentinel surveillance program and to evaluate the malaria prevalence at those points. METHODS: By CRM to compare data on malaria cases reported from national sentinel surveillance program and those from the registration of case report system (CRS). Analysis was made to express the difference regarding the number of cases in order to get the estimates and the incidence rate. RESULTS: The incidence in the relative higher epidemic area, unstable epidemic area and relative lower epidemic area were 1247.30/100000, 171.50/100000 and 46.10/100000, respectively. The rate of miss-reporting malaria cases in these areas were 65.20% ,45.32% and 66.67%, respectively. CONCLUSION: The situation of the malaria cases in the national sentinel surveillance points could be estimated,using the CRM. PMID- 18396667 TI - [Study on the effect of treatment and the status of drug resistance among HIV/AIDS patients treated with highly active antiretroviral therapy in Shandong province]. AB - OBJECTIVE: To understand the effect of highly active antiretroviral therapy (HAART) on AIDS patients, and to explore the prevalence and the impact of HIV-1 drug resistance in Shandong province. METHODS: 2 cross-sectional studies were carried on in 2004 and 2005, to collect data on clinical symptoms and compliance of the AIDS patients with HAART through questionnaire. Informed-consent principle was followed to test on immunological, viral and laboratory index of them. HIV-1 drug genotype resistance by sequencing the gene of HIV-POL after RT-PCR was performed and analyzed. RESULTS: 31 AIDS cases with and. 27 AIDS cases without HAART, were studied. 83.3% and 64.5% of the AIDS patients with HAART showed that the CD4+ T cell count was rising to over 350/microl, in the first study (2004) and in the second (2005) study respectively but still 45.8% and 45.2% of AIDS patients under HAART in the 2 years showed a decreasing HIV load under the detected limit. However, these findings were showing remarkable difference when compared with the AIDS without HAART. 7 drug resistance gene sites were found in AIDS patients with HAART and in AIDS patients without HAART. The rate on high degree drug resistance mutation and total drug resistance rate of mutation of the former were higher remarkably than those of the latter. CONCLUSION: Most of the AIDS patients with HAART met the purpose of rebuilding immunity and control of HIV,as well as alleviation of symptoms. Although the drug resistance stain appeared in Shandong,but had little effect on HAART. AIDS; Drug resistance; Highly active antiretroviral therapy PMID- 18396668 TI - [Study on genotypic resistance mutations to antiretroviral drugs on HIV strains of treated and treatment-naive HIV-1 infectious patients in Hubei province]. AB - OBJECTIVE: To study the drug resistance status on HIV-1 patients who had been treated with highly active antiretroviral therapy (HAART) and those treatment naive ones in Hubei province. METHODS: Nested polymerase chain reaction (PCR) was used to amplify 2 kb DNA fragment in HIV pol gene from peripheral blood of the HIV infected patients and the PCR products were sequenced. The sequences were compared to the Stanford HIV drug resistance database. RESULTS: Nineteen patients were treated with regimens composed of two Nucleoside Reverse Transcriptase Inhibitors (NRTIs) and one Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTI), with 25 patients as treatment-naive. Some protease (PR) drug-resistant mutations were found in these samples, such as D30N (2.27%), D30G (2.27%), M46I (4.55%), M46N (2.27%), I47V (4.55%), I84V (4.55%), I84L (2.27%), N88S (2.27%) and L90S (2.27%) that all belonged to major drug-resistant but A71T (29.55%) belonged to minor resistance mutations Five treated patients were detected having mutations associated RT drug resistance: M41L (5.26%), A62V (5.26%),D67N (5.26%), L210W (5.26%), T215Y (15.79%); K103E (5.26%), K103N (10.53%), Y181C (5.26%), G190A (5.26%), K238N (5.26%), while five treatment-naive patients were detected to have had mutations associated RT drug resistance M184V (4%), K65N (4%), Y115M (4%), F116L (4%), M184I (4%), V179D (4%), G190R (4%).Some additional mutations were detected in RT whose role involve in drug resistance still remained unknown. F214L was positively associated with HAART treatment (P = 0.03). CONCLUSION: Significant differences were found between drug resistance mutations to RTIs in treated and treat-naive patients in Hubei province,indicating that drugs had affected the occurrence of drug resistance mutations. At the same time, novel RT mutations F214L might be associated with HAART or some other drugs. PMID- 18396669 TI - [Detection of serum anti-F antibody in hepatitis C virus infected patients]. AB - OBJECTIVE: To assess the prevalence of serum anti-F in patients with hepatitis C virus (HCV) infection and the distribution of anti-F. METHODS: The recombinant protein (HCV-F/GST) was coated onto micro titer plates as antigen. Sera of 120 patients with hepatitis C virus infection, 15 patients with hepatitis B, 3 patients with hepatitis E and 10 normal sera were tested by indirect ELISA for detecting anti-F. RESULTS: 82 samples out of the 120 (68%) HCV infected patients exhibited a positive anti-F reaction, showing significant difference from the controls with no HCV infection (P < 0.01). Data from logistic analysis showed that the positive rate of anti-F was higher in patients over 50 year olds (OR = 6.675, 95% CI: 2.407-19.071). Patients of midrange, severe phase and hepatic cirrhosis had higher rate than the others (OR = 2.749, 95% CI: 1.470-5.141). CONCLUSION: Prevalence and distribution of anti-F in Yixing hepatitis C patients was reported and which might be related to the progression of HCV infection. PMID- 18396670 TI - [Identification of harboring stx2::IS1203 Escherichia coli O157:H7 strains isolated in China]. AB - OBJECTIVE: To understand the variation of Shiga toxin (stx) genes of Escherichia coli O157:H7 strains isolated in China. METHODS: Polymerase chain reaction (PCR) was used to identity the types of stx genes and the nucleotide sequences of the amplified stex variants genes were determined. Compare to the cytotoxicity of Stx,variants were tested by HeLa cell assay. RESULTS: We found novel stx2 genes in 3 of 289 strains of Shiga toxin-producing E. coli O157:H7 isolated from 1999 to 2002 in China. The novel stx2 genes were inserted by a 1.3-kb insertion sequence (IS) and the nucleotide sequences of IS showed 100% homology with that of IS1203 variant (IS1203v). The IS1203v inserted in the stx2 genes of three E. coli O157:H7 strains at different sites and the direction of the open reading frames (ORFs) of IS1203v of each strain was different. In addition to the above mentioned findings, the nucleotide sequences of three stx2 genes were completely identical and the type of the three Stx2 was Stx2 prototype. Compare to the cytotoxicity of Stx2 prototype, the novel Stx2 was found to be obviously lower. CONCLUSION: E. coli O157:H7 strains harboring stx2::IS1203v genes were isolated in China. Consequently, the results of HeLa cell assay showed that the insertion of IS1203v could lead to low cytotoxicity of Stx2. PMID- 18396671 TI - [Study on the neutralization capacity of different types of human measles virus vaccine and the epidemic strains]. AB - OBJECTIVE: To explore the neutralization capacities of different types of human serum to measles virus epidemic strains and vaccine strain. METHODS: Neutralization antibody (NT) to Shanghai 191 and measles virus isolates in 2005 were tested using acute and convalescent serum samples from diagnosed measles patients, children serum samples collected before and after vaccination and serum samples of migrant residents, from 3 different regions. Additionally, animal immune serum referring to vaccine strain and 3 epidemic strains were prepared and used to undergo crossing neutralization test with corresponding strains mentioned above. Antigenic ratios were calculated. RESULTS: GMT value of NT of after-immune serum to vaccine strains was 50.82,1.86 times higher than that to MVi/ZJ/05/7 (GMT was 27.35), whereas GMT value of convalescent serum to MVi/ZJ/05/7 (GMT was 386.95) was obviously higher than that to vaccine strain (GMT was 1:151.83),and GMT value of migrant residents' serum in 3 regions to MVi/ZJ/05/7 were 2.22-4.17 times lower than that to vaccine strain. Meanwhile,the antigenic ratios between MVi/ZJ/ 99/1, MVi/ZJ/04/1, MVi/ZJ/05/7 and vaccine strain were found to be 4.28,5.24 and 5.66 respectively. Additionally,low NT titers to vaccine strain were found in patients' acute sera and GMT value was over 1:4. CONCLUSION: There were obvious differences on neutralization antibody of different types of serum to measles vaccine strain and epidemic strains which indicating the antigenic diversity of epidemic strains had influenced the protective effectiveness of vaccine antibody to epidemic strains. It was of significance to carry on research projects on the antigenic diversity and effectiveness of measles vaccine. PMID- 18396672 TI - [Study on the prevalence rate of rheumatoid arthritis in Zhuang nationality population from Nanning, Guangxi]. AB - OBJECTIVE: To study the prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi. METHODS: A total of 7407 people with Zhuang nationality, aged 16 and over were surveyed. Questionnaire was administered to each subject under survey, while X-ray and serum rheumatoid factor were carried out to all those who gave positive answers. Physical examinations were done to the suspicious cases by experts on rheumatism. RESULTS: were compared with those in 6826 people of Han ethnicity living in the same district. RESULTS: The prevalence rates of rheumatoid arthritis in Zhuang nationality population was 0.27% when comparing to the Han population which was 0.28% (P > 0.05). Rates of awareness on rheumatoid arthritis in Zhuang and Han population were 5.0% and 10.5% (P > 0.05). After the diagnosis of rheumatoid arthritis was made and among patients who had received the treatment, the rates were 0% vs. 5.25%. CONCLUSION: The prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi was not significantly different than that in Han ethnic group. However, the rates on awareness and the treatment of rheumatoid arthritis were still under satisfaction. PMID- 18396673 TI - [Investigation on the health-related quality of life among 411 patients with irritable bowel syndrome]. AB - OBJECTIVE: To evaluate and compare the health-related quality of life (HRQOL) on patients with irritable bowel syndrome (IBS). METHODS: Following the Rome III Criteria, 411 IBS patients and 430 healthy people were selected as subjects,who were outpatients in Department of Gastroenterology, 2nd Hospital of Xi' an Jiaotong University and Shaanxi Provincial People's Hospital from July 2006 to April 2007. Using the 36-Item Short-Form Health Survey (SF-36). This study compared the SF-36 scale scores of IBS patients with the healthy people. RESULTS: On all of the 8 SF-36 scales, patients with IBS scored significantly lower than healthy people (P < 0.001). Decrements in HRQOL were most predominant in general health perception and role limitations caused by emotional health problem, with scores of 33.5 + 16.9, 40.8 +/- 25.1 respectively. The emotional well-being and energy/fatigue scale scores were also quite low (42.2 +/- 19.3,43.1 +/- 20.2,respectively). They also scored significantly lower on both physical summary. and mental summary scores (P < 0.001). IBS patients were classified into IBS with constipation,IBS with diarrhea, mixed IBS and unsubtyped IBS subgroups, with percentages as 25.3%, 50.1%, 11.2% and 13.4% respectively. CONCLUSION: IBS patients experienced great impairment in HRQOL. These data offered further insight into the impact of IBS on patient functional status and well-being. PMID- 18396674 TI - [Study on the application of Rogerson Spatial Pattern Surveillance Method in real time surveillance for infectious diseases]. AB - Application of Rogerson Spatial Pattern Surveillance Method was discussed in real time surveillance and early monitoring for infectious diseases. Theory of Rogerson Spatial Pattern Surveillance Method was introduced in detail. Data from the program on 'Monitoring epidemic cerebrospinal meningitis with Rogerson Spatial Pattern Surveillance Method' by the Chinese Center for Disease Control and Prevention provided evidence as how to choose crucial parameters such as K, H, n0 and tau, how to simulate surveillance process and how to interpret results. Rogerson Spatial Pattern Surveillance Method seemed to be able to continuously monitor the changes of infectious diseases. New cases could be added to this model at any moment and this model could provide early warning information in time when the excess of cases occurred. Simultaneously it could provide the time and site of the extra cases. Rogerson Spatial Pattern Surveillance Method could implement dynamic and real-time surveillance for infectious diseases. Simultaneously it could avoid the problem of multiple tests that causing large type I error. It greatly helped in providing reliable information for controlling infectious diseases in early epidemic period. Hence,it had important value of application in early warning on infectious diseases. PMID- 18396675 TI - [The progression of methods for estimating the size of populations at risk for acquired immunodeficiency syndrome]. PMID- 18396676 TI - [Knowledge and its influencing factors on HIV/AIDS occupational exposure and self protection among medical personnel of Guangzhou city]. PMID- 18396677 TI - [A case control study of risk factors in HIV infected pregnant women]. PMID- 18396678 TI - [An epidemiological survey on bronchial asthma of Uygur adults in the lowest region of China]. PMID- 18396679 TI - [Parental rearing styles and personality characteristics of young inpatients with internet addiction disorders in different family types]. PMID- 18396680 TI - [Mesenchymal stem cell-mediated immuno-gene therapy for tumors]. PMID- 18396681 TI - [Effect of blocking of ROCK-1, an effector of small G protein Rho, on the malignant behavior of ovarian tumor cells in vitro]. AB - OBJECTIVE: To investigate the possible role of ROCK-1 in ovarian cancer invasion and metastasis. METHODS: ROCK-1 ASODN was transfected into SW626 and Caov-3 cell lines mediated by Lipofectamine 2000. The expressions of ROCK-1 mRNA and protein were detected by RT-PCR and Western-blot assay. Boyden chamber was used to assess the effect of ROCK-1 ASODN on the invasion and migration of the cell lines. The changes in the adhesion and proliferation of the transfected cells were detected by MTT assay. RESULTS: The expressions level of ROCK-1 mRNA and protein in the cell lines were decreased significantly after transfection at doses of 10 micromol/L and 20 micromol/L ROCK-1 ASODN. When compared with the control group, the invasion capability of transfected cells was inhibited to an extent of 75.6% +/- 3.8% and 54.7% +/- 2.9%, respectively, for SW626 cell line, and 68.8% +/- 4.7% and 50.0% +/- 4.5% for Caov-3 cell line, respectively. The random migratory activity of these two cell lines was inhibited by 80.0% +/- 1.3%, 63.7% +/- 1.9%, 72.5% +/- 3.4% and 55.9% +/- 2.5%, respectively, and the inhibition of chemotaxis activity of the two cell lines was 83.9% +/- 1.4%, 64.1% +/- 1.3%, 72.5% +/- 3.4% and 54.5% +/- 1.9%, respectively. No significant difference was found in the adhesion and proliferation of the cells transfected with ROCK-1 ASODN and control cells. CONCLUSION: The expression of ROCK-1 was closely related to the invasion capability and migratory activity of ovarian cancer cells. ROCK-1 may play a crucial role in invasion and metastasis of ovarian cancer. PMID- 18396682 TI - [Immune response of HBsAg gene-modified dendritic cell-based vaccine in HepG2. 2. 15 hepatocellular carcinoma cells]. AB - OBJECTIVE: To study the cytotoxic T lymphocyte (CTL) response induced by dendritic cells (DC) transduced with recombinant adenovirus vector bearing hepatitis B virus surface antigen (HBsAg) gene in hepatocellular carcinoma HepG2. 2. 15 cells in vitro. METHODS: Full length HBsAg cDNAs were subcloned into pIND vector, followed by being cloned into pShuttle vector. The HBsAg gene fragments resulted from the pShuttle-S digested with PI-Sce and I-Ceu were linked to the linear adeno-X virus DNA. After packaged with HEK293 cells, the adenovirus expression vector was obtained. Then the recombinant adenovirus expression plasmid AdVHBsAg was transfected into human monocyte-derived dendritic cells, to construct AdVHBsAg hepatocarcinoma tumor vaccine. The effectiveness of transfection was detected by Western blot. Surface molecules of AdVHBsAg-DC were detected by FACS. Autologous T cell proliferation stimulated by AdVHBsAg-DC was detected by 3H-TdR assay. Cytotoxic CTL activity induced by AdVHBsAg-DC in vitro was detected by LDH assay. RESULTS: HBsAg gene in the inserted DNA of AdVHBsAg was confirmed by PCR, and predictive fragments proved by restriction enzyme digestion analysis were exhibited. Cell pathological changes appear after 10 days HEK293 cells transfected AdVHBsAg. Western blot analysis showed that HBV surface antigen gene was expressed in transfected DC, indicating that the transfection was effective. AdVHBsAg-DC was able to upregulate CD1a, CD11c, CD80, CD86 and HLA DR. Autologus T cell proliferation induced by AdVHBsAg-DCs was significantly higher than that in DC control group and LacZ-DC group (P < 0.05). AdVHBsAg-DC activated CTL presented the specific killer ability to the hepatocellular carcinoma cells expressing HBsAg. CONCLUSION: DC transduced with recombinant adenovirus HBsAg can express HBV-related hepatocellular carcinoma antigen (HBsAg), and AdVHBsAg-DC can induce potent immune response against HBsAg-positive hepatocellular carcinoma cells in vitro. PMID- 18396683 TI - [Killing activity of co-cultured cytokine-induced killer cells and dendritic cells against multi-drug resistant tumor cell lines]. AB - OBJECTIVE: A lot of studies have suggested that a certain amount of T cells may be involved among cytokine-induced killer (CIK) cells. The aim of the present study was to prove whether an antigen-specific killing effect on tumor cells is involved during the CIKs-induced killing process. METHODS: Bone marrow mononuclear cells (BMMNCs) derived from healthy subjects were separately cultured to generate dendritic cells (DC) and CIKs. A human mammary cancer cell line MCF 7/ADR, expressing P-gp antigen, was frozen-thawed and the lysate including P-gp antigen was obtained. The DC pulsed with or without tumor antigen lysate was co cultured with CIK (pulsed-DC + CIK and DC + CIK), and CIK cultured alone was used as control. The cell phenotype of DC and CIK was analyzed by flow cytometry. The secretion of IL-12 and IFN-gamma was assayed by ELSA. The antitumor effect of the three CIK groups targeted at MCF-7/ADR cells expressing P-gp antigen and MCF-7 cells was detected by MTT. RESULTS: Pulsed-DC + CIK group and DC + CIK group showed a higher expression level of DC mature phenotypes than those before co culture with CIK, with a significant difference (P = 0.003, P = 0.001, respectively). The phenotypes (CD3, CD8, CD56) of CIK in pulsed-DC + CIK group and DC + CIK group was higher than those in CIK group (P = 0.003, P = 0.011, respectively). Among the three CIK groups, pulsed-DC + CIK group had the highest phenotypes on CD3+ CD56 (pulsed-DC + CIK vs. DC + CIK, P = 0.001; pulsed-DC + CIK vs. CIK, P < 0.001) and CD3 CD8 (P = 0.002, P = 0.002, respectively). Among the three groups, the pulsed-DC + CIK group showed the lowest CD45RA phenotype (pulsed-DC + CIK vs. DC + CIK, P < 0.001; pulsed-DC + CIK vs. CIK, P = 0.004). Among the three groups the secretion of IL-12 and IFN-gamma had the highest level in pulsed-DC + CIK group, with a value of 254 +/- 14.5 pg/ml and 3100 +/- 286 pg/ml, respectively. The antitumor killing effect on MCF-7/ADR cells had a significant difference between any two groups (pulsed-DC + CIK VS. DC + CIK, P = 0.039; pulsed-DC + CIK VS. CIK, P = 0.002; DC + CIK vs. CIK, P = 0.049). The highest was in pulsed-DC + CIK group and the lowest was in CIK group. The CIK group showed a significantly lower antitumor effect on MCF-7 cells than the other two groups (pulsed-DC + CIK vs. CIK, P = 0.007; DC + CIK vs. CIK, P = 0.048), but no significant difference between the pulsed-DC + CIK and DC + CIK groups. CONCLUSION: In the present study, DC and CIK cells have been successfully obtained and cultured from bone marrow mononuclear cells. After their co-culture, not only both their specific phenotypes were increased, but also the associated cytokines were secreted. An improved antitumor killing effect and some possible specific immunocytotoxicity were observed. Our findings provided a basis for experimental and clinical research on bio-immunotherapy targeted at multi-drug resistant tumor cells. PMID- 18396684 TI - [Expression of annexin I in human pancreatic cancer and the influence of its down regulation on biology of this cancer]. AB - OBJECTIVE: To investigate the expression of annexin in human pancreatic cancer and to elucidate its role in oncogenesis of pancreatic cancer. METHODS: A pancreatic carcinoma cell line Suit-II with high-expression of annexin I gene was adopted. Three subtypes of annexin I -siRNA sequences and a non-related fragment were combined, and the eukaryotic expression vectors bearing siRNA fragments were constructed. Then they were transfected into pancreatic carcinoma cells to knock down the expression of annexin I by RNAi. After knocking down the expression of annexin I , the growth speed, cell cycling, morphological features and apoptosis of pancreatic carcinoma cells were examined by RT-PCR and MTT test. RESULTS: When the expression of annexin I was blocked, the growth speed of pancreatic carcinoma cells was significantly decreased, the morphological features were changed and pronounced apoptosis occurred. CONCLUSION: Annexin I can modulate pancreatic carcinoma cell cycle, promote the cell proliferation, increasingly stimulate the cell growth, and suppress the process of apoptosis in pancreatic carcinoma cells. PMID- 18396685 TI - [Decreased expression of Cx32 and Cx43 and their function of gap junction intercellular communication in gastric cancer]. AB - OBJECTIVE: The aim of this study was to investigate the expression of connexin (Cx) and the function of gap junction intercellular communication (GJIC) in the carcinogenesis, progression and metastasis of gastric cancers. METHODS: Immunohistochemistry was used to detect the expression of Cx32 and Cx43 proteins in tissue samples. Indirect immunofluorescence assay was used to investigate the expression of Cx32 and Cx43 proteins in several gastric cancer lines of various differentiation grades. The expression of Cx43 in samples of gastric cancer tissue, adjacent normal tissue and in the gastric cancer cell lines of various differentiation grades was detected by Western blot. Scrape-loading dye transfer (SLDT) technique was used to detect the function of gap junction intercellular communication (GJIC) in the various cell lines. RESULTS: In the normal gastric mucosa the expression rates of both Cx32 and Cx43 were 100%. In gastric cancers, the expression rates of Cx32 andCx43 were 49.5% (55/111) and 39.6% (44/111), respectively. There was a significant difference between their expression in normal and cancer tissues (P < 0.05). Age of the patients was not significantly correlated with the expression level of Cx32 and Cx43 (P > 0.05). Cx43 expression was significantly associated with the TMN stage, histological type, depth of infiltration and distant metastasis (P < 0.05), but Cx32 expression was not significantly correlated with depth of infiltration ( P > 0.05). In the cancer cell lines, a positive expression of Cx32 and Cx43 was detected in transfected human stomach mucosal cell line (CES-1) and human well differentiated stomach cancer cell line (N87), but negative in the poorly differentiated stomach cancer cell line (BGC-823) at all. Both Cx32 and Cx43 expression rates were 100% in the cell line GES-1. Cx32 expression rate was 49.0% and Cx43 expression rate was 55.0% in the cell line N87. But in the poorly differentiated cancer cell line BGC 823 both Cx32 and Cx43 expression was negative. GJIC function detection showed: GES-1 showed well GIJC function but no GIJC function in the cell lines N87 and BGC-823. The intensity of fluorescence was gradually decreasing from GES-1 cells to N87 cells and almost no fluorescence in BGC-823 cells. Western blotting showed that Cx43 expression in normal tissue was higher than that in gastric cancer tissue, and in the cell lines GES-1, N87 and BGC-823, the bands seemed decreasing progressively. There was very low expression in BGC-823 cells. CONCLUSION: The decreasing expression of connexin Cx32 and Cx43 is obviously correlated with the occurrence, development and metastatic potential of stomach cancers. PMID- 18396686 TI - [Prognosis of locally advanced non small cell lung cancer treated with three dimentional conformal radiotherapy]. AB - OBJECTIVE: To summarize our experience and evaluate the prognostic factors of locally advanced non small cell lung cancer (LA-NSCLC) treated with three dimentional conformal radiotherapy (3D-CRT). METHODS: 118 patients with stage IIImA/IIIB non small cell lung cancer were treated with 3D-CRT from Nov. 2001 to Mar. 2005. 113 patients with complete clinical data were eligible for analysis, 45 of them received radiotherapy alone; 39 were treated by concurrent chemoradiation with paclitaxol plus carboplatin in 32 patients and topotecan in 7 patients, and 29 by sequential chemoradiation with platinum-based regiment in most of them. The dose of radiation for the thoracic field ranged from 26 Gy to 75 Gy with a median dose of 60 Gy. GTV and PTV were collected from the 3D treatment plans in 79 and 101 patients, respectively. Overall survival (OS) was calculated using the Kaplan-Meier method. Comparisons among the curves were made using a two-tailed long-rank test. The Cox model was used for multivariate analysis. RESULTS: The 1-, 2- and 3-year overall survival rate was 60.7%, 31.6% and 22.4%, respectively, with a median survival time of 17 months. In univariate analysis, the following characteristics were significantly associated with longer survival: absence of chest pain, good karnofsky performance status (KPS), albumin > 4.2 g/L, hemoglobin > or = 140 g/L (male) or 130 g/L (female), response to radiotherapy and GTV < 100 cm3. However, multivariate analysis revealed that only good KPS was an independent risk factor predicting the survival. CONCLUSION: Three-dimensional conformal radiotherapy is effective in the treatment of locally advanced non-small cell lung cancer with acceptable complications. Karnofsky performance status is the only independent prognositic factor. PMID- 18396687 TI - [MRI in exploring the spread pattern of retropharyngeal lymp node metastases in nasopharyngeal carcinoma]. AB - OBJECTIVE: The purpose of this study is to investigate the incidence, distribution and spread pattern of retropharyngeal lymph node (RLN) metastasis in the patient with nasopharyngeal carcinoma (NPC) using magnetic resonance imaging (MRI). METHODS: MRI data of 294 patients with newly diagnosed NPC were reviewed retrospectively. Criteria for RLN metastasis were the minimal axial dimension of > or = 6 mm, or the presence of nodal necrosis. RESULTS: RLN involvement was detected in 165 (56.1%) patients. The incidence of metastatic lateral RLN decreased gradually from C1 to C3 level. A significantly higher incidence of metastatic RLN was observed in the patients with cancerous involvement of oropharynx, nasal cavity, pre-styloid parapharyngeal space, post-styloid parapharyngeal space or with cervical lymph node metastasis. Thirty-one patients (10.5%) had metastatic RLN only, and 54 (18.4%) showed metastatic cervical lymph nodes only, but 134 (45.6%) exhibited an involvement of both the RLN and cervical lymph node. A significantly lower incidence of metastatic RLN was found in T1, N0, and stage I disease. CONCLUSION: The incidence of metastatic lateral retropharyngeal lymph node decreases gradually from C1 to C3 level. Metastatic retropharyngeal lymph node is associated with cancerous involvement of surrounding tissue by primary tumor or with cervical lymph node metastasis. Retropharyngeal lymph node or cervical level II node can be considered equally as the first station of nodal metastasis in nasopharyngeal carcinoma. PMID- 18396688 TI - [Expression of epidermal growth factor receptor and the correlation with HPV16/18 infection in cervical intraepithelial neoplasia and cervical carcinoma]. AB - OBJECTIVE: The aim of this study was to elucidate the role of EGFR expression and HPV infection and their relationship in the genesis and progression of cervical carcinoma. METHODS: This analysis included 60 cases of cervical carcinoma, 40 cases of CIN and 30 cases of control group. Patients of cervical carcinoma group were all subjected to radical hysterectomy with bilateral pelvic lymphadenectomy in Tumour Hospital, Zhongshan University from 1997 to 2001. The FIGO stage was I a - II b. EGFR protein was stained by S-P immunohistochemistry, and HPV16/18 DNA was detected by PCR. RESULTS: The moderate/ strong expression of EGFR was observed in 0, 42.5%, 76.7% of normal cervical epithelium, CIN and cervical tumor tissue, respectively, with a significant difference among them (P < 0.05). The infection of HPV16/18 was observed in 6.7%, 67.5%, 58.3% of normal cervical tissue, CIN and cervical tumors, respectively. The infection rate of CIN or cervical carcinoma was significantly higher than that in normal cervicaltissue (P = 0.000), but no statistically significant difference was observed between cervical carcinoma and CIN. The moderate/strong expression of EGFR demonstrated an association with the cervical invasion depth. The EGFR expression increased significantly as the invasion depth progressed from less than or equal to a half cervical stroma to deeper than that (89.2% vs. 56.5%, P = 0.004). The infection of HPV16/18 demonstrated a correlation with the cervical canal invasion. The infection increased significantly in the cases with cervical invasion than that in the cases without invasion (88.2% vs. 46.5%, P = 0.003). But no significant correlation was observed between EGFR and HPV. Neither EGFR nor HPV had a significant association with carcinoma prognosis. CONCLUSION: EGFR and HPV demonstrate a significant correlation with genesis and progression of cervical carcinoma. In our study, neither EGFR nor HPV demonstrates a significant association with tumor prognosis, and no significant correlation is observed between EGFR and HPV. PMID- 18396689 TI - [Descriptive analysis of incidence and time trends of esophageal and gastric cancers in Linzhou city]. AB - OBJECTIVE: To analyze the incidence and time trends of esophageal and gastric cancers in Linzhou city bassed on the data of Linxian Tumor Registry, and to provide valid reference data for research and effective estimation of cancer control in this area. METHODS: All incidence records for the both cancers during 1988-2003 were drawn from Linzhou Tumor Registry and grouped by sex, age, year and then linked to corresponding population data. The incidence rates of those two topographic site cancers were calculated and the age-adjusted rates were calculated by direct standardization to the world population. A joinpoint model was used to get the annual percentage change (APC) of the age-adjusted rates, and to estimate the epidemiological trends of both cancers in population of Linzhou city. RESULTS: In the year 2003 the age-adjusted incidence rates of esophageal and gastric cancers were 81.78 per 100 000 and 77.08 per 100 000, respectively, in the population of Linzhou city. The incidence rate of both cancers showed a decreasing trend from 1988 to 2003. The APC of the incidence rates of esophageal cancer was - 2.6% and that of gastric cancer was - 1.8%, and both indexes were statistically significant (P < 0.05). CONCLUSION: The incidence rates of esophageal and gastric cancers have presented a decreasing trends in the population of Linzhou city. This trend will continue along with the development of social economy, elevation of living standard and improvement in living habit and environment. PMID- 18396690 TI - [Expression and significanc of ERbeta and HER2 in breast cancer]. PMID- 18396691 TI - [Weekly paclitaxel and carboplatin with concurrent three dimensional conformal radiotherapy for locally advanced non small cell lung cancer]. AB - OBJECTIVE: To evaluate the toxicity and efficacy of weekly chemotherapy of paclitaxel and carboplatin with concurrent three dimensional conformal radiotherapy (3D-CRT)for locally advanced non small cell lung cancer(NSCLC). METHODS: From July 2002 to July 2004, a non-randomized prospective study of 52 patients with locally advanced NSCLC treated with 3D-CRT and chemotherapy by paclitaxel and carboplatin were carried out. Of the 52 patients, 21 received concurrent chemoradiation with 3D-CRT and weekly paclitaxel and carboplatin (concurrent chemoradiation group), 31 received sequential chemoradiation with paclitaxel and carboplatin and 3D-CRT (sequential chemoradiation group). In the concurrent chemoradiation group, paclitaxel 40 mg/m2 was administered intravenously for 1 hour. Carboplatin of 1.5 AUC/cycle was used after administration of paclitaxel on D1, D8, D15, D29, D36 and D143. In the sequential chemoradiation group, two cycles of chemotherapy were given at two weeks before radiotherapy. Paclitaxel 150 mg/m2 and carboplatin 5 AUC/cycle were administered on D1 and D21. 3D-CRT was given at two weeks after the second cycle of chemotherapy provided that the hematological examination was normal. 3D-CRT was given at 1.8-2.0 Gy/f to a total dose of 60-66 Gy/6-8 weeks. RESULTS: All the patients completed the trial, but 12 had prolongation of treatment time for more than 1 week due to severe leucopenia with 5 in concurrent chemoradiation group and 7 in sequential chemoradiation group. The rate of complete response (CR), partial response (PR), progress of disease (NC + PD) and overall response was 9.5% (2/21), 71.4% (15/21), 19.0% (4/21) and 81.0%, respectively, in concurrent group, versus 6.5% (2/31), 67.7% (21/31), 25.8% (8/31) and 74.2% in sequential group (CR), respectively. II-III grade of esophagitis, pneumonia and leukocytopenia observed in concurrent chemoradiation group was 61.9% (13/21), 41.9% (13/31) and 23.8% (5/21) ,versus 22.6% (7/31), 42.9 % (9/21) and 19.4% (6/31), respectively, in the sequential chemoradiation group. One of those patients in concurrent chemoradiation group had IV grade of leukocytopenia. The overall median survival time was 17.5 months with 19.0 months for concurrent chemoradiation group, 15.8 months for sequential chemoradiation group. The Overall 1-, 2-year survival rate was 72.0%, 37.0%, respectively and 1 - and 2 - year local control rate was 75.0% and 75.0%. CONCLUSION: Our data indicate that concurrent chemoradiotherapy is safe and effective for locally advanced non small cell lung cancer. Concurrent chemoradiotherapy may be helpful in improving response and survival than sequential one, but no significant difference is observed between two groups in this series(P > 0.05). Further randomized prospective study is still needed to prove it. PMID- 18396692 TI - [Concurrent chemoradiation with paclitaxel and platinum for locally advanced esophageal cancer]. AB - OBJECTIVE To assess the efficacy of concurrent chemoradiation with paclitaxel and platinum and external irradiation, and to compare the effect of extensive regional field irradiation with conventional local field irradiation for locally advanced esophageal cancer. METHODS: From Oct. 2000 to Jan. 2006, 89 patients with locally advanced esophageal cancer were registered in this study. All patients were inoperable or refused to undergo operation. Patients were divided into two groups: extensive regional field group (51 patients) and conventional field group (38 patients). Patients received radiotherapy at a total dose of 60 Gy in 30 fractions within 7 weeks,and concurrent paclitaxel 125 mg/m2 on D1, cisplatin 20 mg/m2 on D1-D3, or oxaliplatin 130 mg/m2 on D2 in the fist and fourth week of external radiation. RESULTS: Of these patients, 87.6% completed the treatment regimen with a response rate of 75.5% and 66.7% in the extensive regional field group and conventional field group, respectively. Grade 3 or severe toxicities of leucopenia (33.3% vs. 23.7%), thrombocytopenia (76.0% vs. 2.6%), and esophagitis (17.7% vs. 26.3%) were observed in extensive regional field group and conventional field group, respectively. Major late toxic effect was lung fibrosis. There were no statistically significant differences in the incidence of the toxicity profile between two groups. The overall 3-year survival rates was 32.8%, and the overall 3-year recurrence and metastasis-free survival rates was 34.5%. The overall 3-year locoregional control rate was 44.0%. No significant difference was found between two groups in the 3-year survival (38.2% % vs. 28.1%, P = 0.59). For the patients with stage II and stage III cancers who completed the planned treatment, large regional field radiotherapy significantly improved the 3-year survival (57.3% vs. 22.2% , P = 0.03) or 3-year recurrent and metastasis-free survival (55.5% vs. 23.0%, P = 0.03) or 3-year locoregional control (65.9% vs. 30.2%, P = 0.02) than conventional field radiotherapy. CONCLUSION: historical results, the combination of paclitaxel/platinum and radiation in this study can improve the survival for locally advanced esophageal, and the side effect is well tolerated. Compared with the conventional field group, concurrent chemoradiotherapy with the large regional field can significantly improve 3-year survival and locoregional control for stage II or stage III esophageal cancer. PMID- 18396693 TI - [Clinical features and prognosis of alveolar soft part sarcoma]. AB - OBJECTIVE: To investigate the clinical feature, suitable treatment and prognosis of alveolar soft part sarcoma. METHODS: The clinical data of 58 such patients treated at our hospital from 1967 to 2006 were retrospectively analyzed. Fifty three patients except 6 with distant metastasis underwent surgical resection including 33 extensive resection and 19 local resection. Furthermore, 19 patients received postoperative adjuvant radiotherapy or chemotherapy. RESULTS: Eleven of 50 patients who underwent complete resection developed local recurrence. Thirty One (53.4%) was found to have lung metastasis. The overall 3-, 5-, 10-year survival rate was 89.5%, 74.1% and 57.7%, respectively. The median survival time of the patients was 125 months. The 3-, 5-, 10-year survival rate was 100.0%, 81.6% and 65.3% for female patients, but it was 79.6%, 67.2% and 49.7% for male patients, respectively. CONCLUSION: Alveolar soft part sarcoma usually grows slowly. Though local recurrence is rarely seen, distant metastasis is frequently observed. Lung is the most common organ of metastasis. However, survival can still be long even with lung metastasis. To achieve microscopically complete resection is quite critical to localized alveolar soft part sarcoma. No survival advantage was observed when adding adjuvant radiotherapy and/or chemotherapy. The phenomena of female patient or patient with X chromosome translocation showing better prognosis than the male or other patients needs further investigation. PMID- 18396694 TI - [Clinical analysis of primary small intestinal neoplasms in 305 cases]. AB - OBJECTIVE: To summarive the experience in diagnosis and treatment of primary small intestinal neoplasm. METHODS: The data of 305 patients with pathologically confirmed primary small intestinal tumor collected from 6 hospitals around the Songhua River during the past 33 years were analyzed retrospectively. RESULTS: There were 42 benign and 263 malignant tumors in this series with a ratio of 1: 6.26. The 263 malignant tumors in this series consisted of 135 adenocarcinomas, 57 malignant stromal tumors, 37 malignant lymphomas, 20 carcinoids, and etc. Chronic occult bleeding, gradual of body weight loss and mild abdominal pain (three obscurities) were the common clinical features and alerting massage of intestinal tumor. Correct preoperative diagnostic rate was only 57.0% (174/305) due to difficulty in early diagnosis, which was 67.2% (92/137) in the duodenal tumors, and 51.9% (82/168) in the jejunoileal tumors. All of the 42 benign tumors were resected completely. For the 263 patients with malignant tumors, radical dissection was performed in 153, palliative resection in 34, and gut by-pass or biopsy in 76. The median survival of the patients who underwent radical resection of their malignant tumors was 92 months, which was significantly higher than that of the other groups. CONCLUSION: Early diagnosis of primary small intestinal tumors is difficult and with a preoperative misdiagnosis rate of 43.0%. Total intestinal barium swallowing, endoscopy and superior mesenteric arteriography are three critical examinations for diagnosis and location. Early surgical resection is crucial in improving the prognosis. The primary small intestinal tumor should be resected as early as possible if no distant metastasis is detected. PMID- 18396695 TI - [Prognostic analysis of 88 patients with ovarian clear cell carcinoma]. AB - OBJECTIVE: To investigate the clinical characteristics of clear cell carcinoma of the ovary and to compare the survival of the patients treated by three different chemotherapy regimens. METHODS: Between 1984 and 2005, the clinical data of 88 surgically treated patients with clear cell carcinoma of the ovary were retrospectively analyzed. Of the 88 patients, 55 (62.5%) had tumor in stage I, 2 in stage II, 22 in stage II, 3 in stage IV and 6 in indefinite stage. These patients underwent either bilateral salpingo-oophorectomy with hysterectomy and omemtectomy or cytoreduction surgery. Of 55 stage I patients, 20 received pelvic lymohadenectomy. All patients were given postoperative chemotherapy, 43 patients received CAP/CP, 33 paclitaxel combination with carboplatinum/cisplatin (TC/TP) and 12 CPT-11 plus MMC. RESULTS: The response rate, recurrence rate, 3- and 5 year survival was 35.0%, 30.2% (13/43), 67.4% (29/43), 43.9% and 29.3%, respectively in patients treated with CAP/CP; 73.9%, 18.2% (6/33), 45.5% (15/33), 57.3% and 40.5%, respectively in the patients with TC/TP; 71.4%, 16.7% (2/12), 25.0% (3/12), 70.7% ( 3-yr survival, no available 5-yr survival), respectively in the patients with CPT-11 + MMC (P < 0.05). During follow-up, 47 (53.4%) patients were found to have recurrence, it was 45.4% (25/55) in stage I patients including 29.6% (8/27) in stage I a + I b and 60.7% (17/28) in stage I c, 75.0% (18/24) in stage II + III and 4/6 in the indefinite FIGO stage. The recurrences rate was 27.8% (5/18) in stage I patients with pelvic lymphadenectomy vs. 51.3% (19/37) in those without. It was 67.3% in 46 patients with elevated CA125, and 38.1% in the other 42 patients with normal or unavailable CA125 (P < 0.05). The overall 3- and 5-year survival rate of 88 patients was 48.7% and 40.9% , respectively, with 72.5% and 66.8% in stage I, 100.0% and 70.5% in stage Ia + Ib, 68.5% and 60.3% in stage Ic, 41.8% and 20.8% in stage II + III, 0 in stage IV (P < 0.05). The 3- and 5-year survival in stage I with pelvic lymphadenectomy was 88.5% and 75.8% vs. 70.3% and 65.1% in those without (P < 0.05). The 3- and 5-year survival of the patients with optimal (residual disease less than 2 cm) was 36.7% and 23.1% vs. 22.2% and 0 in those with suboptimal cytoreduction (P < 0.05), it was 46.8% and 38.8% in the patients with elevated CA125 vs. 46.7% and 43.5% in those with normal one (P > 0.05). CONCLUSION: Our data show that ovarian clear cell cancer patient have a poor response to CAP/CP and may have a better response to TC/TP, especially to CPT-11 plus MMC. However, the overall prognosis is still poor and further clinical investigations are needed to improve it. PMID- 18396696 TI - [Analysis of therapeutic result and prognostic factor in primary fallopian tube carcinoma]. AB - OBJECTIVE: To investigate the impact of treatment modality and clinicopathologic profile on prognosis in primary fallopian tube carcinoma. METHODS: The data of 64 cases with primary fallopian tube carcinoma treated between January 1991 and June 2006 were analyzed. The clinicopathological data were retrospectively analyzed. RESULTS: The overall 5-year survival rate of this series was 56.3%. The overall 3 and 5-year survival rate was 84.6% and 65.4% in surgical staging group versus 58.3% and 33.3% in no surgical staging group with a significant difference between two groups (P = 0.0429; P = 0.043), which was 89.5% and 68.4% in optimal cytoreduction group versus 66.7% and 41.7% in suboptimal cytoreduction group (P = 0.0466; P = 0.0444). However, there was no significant difference in 3-year and 5 year survival rate between the group with pelvic lymphadenectomy and the group without (84.2% vs. 69.2%, P = 0.4667; 63.1% vs. 53.8%, P = 0.459), and also between the group treated using CAP/CP regimen and the group by TP regimen for chemotherapy (81.8% vs. 80.0%, P = 0.8946; 59.1% vs. 60.0% P = 0.9582). It was found that the 5-year survival was correlated with FIGO stage (III-IV vs. I - II, P = 0.0197), differentiation grade (G3 vs. G1 + G2, P = 0.003), pathologic type (other type vs. serous, P = 0.0494), lymph nodes status (positive vs. negative, P = 0.0295). CONCLUSION: Surgical staging, optimal cytoreduction, differentiation grade, pathologic type, lymph node status are important factors influencing the 5 year survival in primary fallopian tube carcinoma. Pelvic lymphadenectomy is necessary and feasible to perform during the procedure of surgical staging and cytoreduction. CAP/CP and TP regiment are similarly effective in adjuvant chemotherapy for primary fallopian tube carcinoma. PMID- 18396697 TI - [Application of exptracorporeal circulation in surgical treatment of abdominal malignancies with tumor thrombus extending into inferior vena cava]. PMID- 18396698 TI - [Essential experience in surgical treatment of esophageal cancer in China over the past five decades]. PMID- 18396699 TI - [Multiple bioimaging modalities in evaluation of an experimental osteonecrosis model induced by a combination of lipopolysaccharide and methylprednisolone]. AB - OBJECTIVE: The present study employed both static and dynamic imaging modalities to study both intra- and extravascular events attributing to steroid-associated osteonecrosis (ON) using an experimental protocol with a single low-dose lippolysaccharide (LPS) injection and subsequently three injections of high-dose methylprednisolone (MPS). METHODS: Fourteen 28-week-old male New Zealand white rabbits received one intravenous injection of LPS (10 microg/kg). After 24 hours, three injections of 20 mg/kg of MPS were given intramuscularly at a time interval of 24 hours. Additional 6 rabbits were used as controls. Dynamic MRI was performed on bilateral femora for local intraosseous perfusion before and after LPS injection. Blood samples were collected for haematological examinations before and after LPS injection. Bilateral femora were dissected and decalcified for microCT-based microangiography. ON lesion, intravascular thrombus and extravascular marrow fat cell size were examined histopathologically. RESULTS: Intravascular thrombus was observed in all ON rabbits. Extravascular marrow fat cell size was significantly increased in ON rabbits than that of the controls (P < 0.05). Compared to baseline, a significant decrease in ratio of tissue-type plasminogen-activator/plasminogen-activator inhibitor 1, activated-partial- thromboplatin-time, and a significant increase in ratio of low-density lipoprotein/high-density-lipoprotein were only found in ON rabbits (P < 0.05). Dynamic MRI showed a significant decrease in the perfusion index 'maximum enhancement' in the ON rabbits (P < 0.05) and microCT-based microangiography showed blocked stem vessels in ON samples. Overall, 93% of the rabbits (13/14) developed ON and no rabbits died throughout the experiment period. CONCLUSION: Both intra- and extravascular events were found attributing to the steroid associated ON based on our experimental protocol with a single low-dose LPS injection and subsequent three injections of high-dose MPS. Both high ON incidence and no mortality in rabbits treated with this inductive protocol suggested its effectiveness for future studies on evaluation of therapeutic efficacy of interventions developed for prevention of steroid-associated ON. PMID- 18396700 TI - [Experimental study on avascular necrosis of femoral head induced by methylprednisolone combined with lipopolysaccharide in rabbits]. AB - OBJECTIVE: To establish a stable animal model for glucocorticoid-induced avascular necrosis of femoral head in rabbits. METHODS: Thirty-six adult New Zealand rabbits were randomly divided into four groups: ten were injected twice with lipopolysaccharide (group A), ten were treated with a combination of lipopolysaccharide and methylprednisolone (group B), ten were injected three times with methylprednisolone (group C), and six were injected normal saline as a control (group D). MR imaging was performed in the rabbits before the first injection of lipopolysaccharide or methylprednisolone, and at 2, 4, and 6 weeks after the last injection of lipopolysaccharide or methylprednisolone. Histopathological changes in the femoral heads were observed by light microscope and transmission electron microscope at the end of six weeks after the injection. Vascular infusion with Chinese ink was made to evaluate the morphological changes of blood vessels in the femoral head. The percentage of trabecular bone area and empty lacunae and microvascular density were measured. According to the histological and MR imaging appearance of the femoral heads in all groups, the incidence of osteonecrosis of every group was calculated. RESULTS: Listlessness, blepharal hyperemia, less activity and reduced diet were found in the rabbits of groups A and B after injected with lipopolysaccharide. At 3 weeks after the final injection, the body weight of groups B and C was decreased. At 4 weeks after the final injection, the body weight of groups A and D was increased. No abnormal signal could be detected on MR images in rabbits of all groups before injection and at 2 weeks after the injection. At 4 weeks and 6 weeks after the last injection, irregular low signal on T1-weighted images and irregular low or highsignal on T2-weighted images could be detected on MR images in rabbits of groups B and C, no abnormal signal could be detected on MR images in rabbits of groups A and D. At 6 weeks after the last injection, the trabecular bone of group B became thin and sparse, some were broken. The percentages of empty lacunae were 11.8% +/- 4.7%, 34.4% +/- 6.2%, 20.0% +/- 4.7% and 9.3% +/- 4.6%; the percentages of trabecular bone area were 59.2% +/- 6.8%, 40.1% +/- 6.0%, 51.5% +/- 5.6% and 63.2% +/- 8.3%; and the microvascular densities were 14.3% +/- 2.7%, 4.5% +/- 2.1%, 10.2% +/- 3.1% and 15.4% +/- 4.1% in groups A, B, C and D respectively. There were statistically significant differences between group B and groups A, C, D (P <0.01). The fatty tamponade accumulated in the medullary cavity and intramedullary vascular sinusoids were pressed by the lipocytes and became narrow. Limposomes were found in osteocytes and vascular endothelia of group B and group C. Osteocytes of group B crimpled and pyknosis or karyolysis of chromatin were observed in these osteocytes, nuclear membrane of the osteocytes was discontinous. Vascular endothelia became swollen and the cell junctions widened or were destroyed in groups A and B. The incidence of osteonecrosis in group B (88.9%) was higher than that in group C (22.2%, P < 0.05). There was no osteonecrosis occurred in groups A and D. Conclusion Methylprednisolone combined with lipopolysaccharide can induce typical rabbit model for early avascular necrosis of femoral head. PMID- 18396701 TI - [An experimental osteonecrosis of femoral head induced by a combination of a single low-dose lipopolysaccharide and methylprednisone]. AB - OBJECTIVE: To investigate both incidence and mechanism attributing to steroid associated osteonecrosis of femoral head (ONFH) using an experimental protocol with a single low-dose lipopolysaccharide (LPS) injection and subsequently three injections of high-dose methylprednisolone (MPS). METHODS: Twenty-five New Zealand white rabbits with body weight of (3.0 +/- 0.3) kg were divided randomly into 2 groups. In treatment group, 19 rabbits received one intravenous injection of LPS (10 microg/kg); 24 hours later, three injections of 20 mg/kg of MPS were given intramuscularly at an interval of 24 hours. Additional 6 rabbits which received normal saline injection at the same time point were used as controls (control group). The blood samples were collected for hematological examinations before and after LPS injection, MRI was performed on bilateral hip six weeks after last MPS injection, meanwhile, bone marrow was aspirated from femoral head region to evaluate stem cell's activity. Bilateral femoral heads were harvested to make histopathology examination. RESULTS: All animals survived throughout the experiment period except one death on the second day after LPS injection. In the histopathological examination for the femoral head, ONFH+ was observed in 16 rabbits (88.9%), and the lesions were mainly in the metaphysis. In ONFH+ rabbits, micro vessels fibrous thrombosis and extravascular marrow fat cell size increasing were found around necrotic bone; The femoral heads of control group had no changes. MRI accurate ratio was 93.8% (15/16). Compared to baseline, a significant decrease in ratio of tissue plasminogen activator/plasminogen activator inhibitor 1 and activated partial thromboplatin time, and a significant increase in ratio of low-density lipoprotein/high-density lipoprotein were only found in ONFH+ rabbits (P < 0.05). Meanwhile there was a significant decrease in the number of CFU-F (8.50 +/-9.63) compared with the control (70.17 +/- 7.78, P < 0.05). CONCLUSION: A single low-dose LPS injection and subsequent three injections of high-dose MPS is effective on building steroid-associated ONFH model, coagulation and lipometabolism abnormality, activity degeneration of stem cell may be the key factors of ONFH. PMID- 18396702 TI - [Preliminary study on cryosurgery for osteonecrosis of femoral head in three-foot canine model]. AB - OBJECTIVE: To establish a animal model of osteonecrosis of femoral head in canine like human. METHODS: The thermal field of canine's femoral head was three dimensionally analyzed with fluent 6.2 software so that the best cryosurgery patent could be designed to maximize the osteonecrosis and minimize extra surgery trauma with the cryosurgery system invented by Shanghai Jiaotong University. Liquid nitrogen was pressurized to 0.5 MPa, poured into femoral head for 6.5 minutes, rewarming to 2 degrees C for 5 minutes and then repoured into it again for another 6.5 minutes. Ten three-foot canines were conducted as the animal models of osteonecrosis of femoral head according to the method above. At the end of follow-up, the results were reviewed by radiologic and pathologic check. Two dogs were conducted as control group. RESULTS: In the experimental group, one of the ten canines was testified to occur osteonecrosis of femoral head after one week pathologically, cell death and vessel breakage of cavitas medullaris in the femoral head was obvious under microscope; in other nine canines being still under follow-up, five with three-month follow-up at least progressed to the collapse of femoral head like human (Ficat III). In control group, no osteonecrosis was found. CONCLUSION: Cryosurgery for osteonecrosis of the femoral head in three-foot canine model may become a method to establish the animal model of osteonecrosis of femoral head like human. PMID- 18396703 TI - [Changes in bone micro-architecture and bone mineral density following experimental osteonecrosis of femoral head by local injection of ethanol in canines]. AB - OBJECTIVE: To measure the changes of bone mineral density and bone micro architecture of the femoral head that harvested from the three-foot bearing ethanol destroyed canine model for osteonecrosis of femoral head, and discuss the influences of local injection of ethanol and biomechanical loading to the structural properties of the femoral head. METHODS: Twenty-four Beagles were divided randomly into four-foot bearing canines and three-foot bearing canines. One fore-limb was fixed randomly in three-foot bearing canines. Osteonecrosis was induced in all experimental animals by local injection of 5 mL pure ethanol into one side of the femoral head. The hind limbs injected with NS were acted as control group, that of three-foot canines injected with ethanol were acted as three-foot canine group, and that of four-foot canines injected with ethanol were acted as four-foot canine group. The contralateral femoral head was injected into equal amount of NS. Animals were sacrificed at the time intervals of 1, 3, 6, and 12 weeks after the injection of ethanol. Quantitative microcomputed tomography was used to characterize changes in bone micro-architecture and bone mineral density of femoral head. RESULTS: The clear three-dimensional model of trabecular bone of necrotic femoral head were obtained. There were no significant differences among 3 groups according to the time line by 1 week after ethanol injection(P > 0.05). At 3 weeks after injection of ethanol, in three-foot canine group and four-foot canine group, the volume of BMC, BMD, BVF, and BS/BV increased gradually as the distance to the drilling canal increased. There were significant differences between 3 regions (P < 0.05). At 6 weeks, in three-foot canine group and four-foot canine group, the volume of BMC, BMD,BVF, and Tb.N of region I and II decreased significantly compared with region III (P < 0.05). At 12 weeks, there are no differences among 3 groups (P > 0.05). There were significant decreases in BMD values, BVF, BS/BV, Tb.N, Tb.Sp and Tb.Th after the injection of pure ethanol. And, the changes were more and more obvious by the time line. These changes were differentiable at 3 weeks after injection of ethanol, and became obvious at 6 weeks. These changes were more obvious at the part that near the injection canal. The changes in three-foot canine group were more obvious than that in four-foot canine group. CONCLUSION: Resorption of necrotic compact bone trabecular may weaken the structural properties of the femoral head. Moreover, remodeling and repairing process of necrotic bone trabecular may be hampered by constant biomechanical loading that presented in three-foot bearing canines, and thereby further weaken the structural properties of the femoral head. Biomechanical loading may be one of the critical reasons that lead to the collapse of femoral head. PMID- 18396704 TI - [An experimental study on treatment of steroid-associated femoral head necrosis with simvastatin and BMSCs transplantation]. AB - OBJECTIVE: To approach the possibility of combination of simvastatin and BMSCs transplantation for steroid-associated osteonecrosis of femoral head. METHODS: The BMSCs harvested from 24 rabbits were prepared for cell suspension at a concentration of 1 x 107/mL, and combined with gelatin sponge. Seventy New Zealand white rabbits received one intravenous injection of lipopolysaccharide (10 microg/kg). After 24 hours, three injections of 20 mg/kg of methylprednisolone were given intramuscularly at a time interval of 24 hours. Forty-eight rabbits diagnosed as having femoral head necrosis by MRI were divided into 4 groups randomly, group A: no treatment; group B: only decompression; group C: decompression and BMSCs transplantation; and group D: simvastatin drench (10 mg/kg.d) decompression and BMSCs transplantation. The general information of animals were recorded; after 4 and 8 weeks of operation, 6 rabbits of each group were chosen randomly to do MRI scan, and femoral heads were harvested to do histopathology and scanning electron microscope examination. RESULTS: After 8 weeks, rabbits became more active than before treatment, and walking way became normal gradually in groups C and D. Four weeks after operation, the MRI low signal region of all groups had no obvious changes, but 8 weeks later, the necrosis signal region of group A magnified while it reduced obviously in group D. Histopathological observation: 4 weeks after operation, diffuse presence of empty lacunae and pyknotic nuclei of osteocytes were found in the trabeculae, and few newborn micrangium could been seen in group A; lots of empty lacunae and a small quantity of newborn micrangium could been found in group B; and large amounts of osteoblats and newborn micrangium were found around the necrosis regions in groups C and D. The positive ratio of empty lacunae and microvessel density in group D were 19.30 +/- 1.52 and 7.08 +/- 1.09, showing significant difference compared with other groups (P < 0.05). After 8 weeks of treatment, the bone trabecula collapsed in many regions in group A; there was fibra callus formation along the decompression channel in group B; few empty lacunae was in the bone trabecular, but the shape of marrow cavity was not normal in group C; and it showed almost normal appearance in group D. The positive ratio of empty lacunae and microvessel density in group D were 11.31 +/- 1.28 and 12.37 +/- 1.32, showing significant differences compared with other groups (P < 0.05), meanwhile, showing significant difference compared with that of 4 weeks after operation(P < 0.05). Scanning electron microscope: 8 weeks after operation, the bone trabecula collapsed in many regions, and few osteoblasts could be found on the surface, a great quantity of fat cells cumulated in the bone marrow in group A; cracked bone trabecula could be found occasionally in group B; the density of bone trabecula was lower than the normal in group C; and the shape of the marrow cavity and the density of bone trabecula were similar to the normal in group D. CONCLUSION: Simvastatin can promote the differentiation of osteocyte and vascular endothelial cell from MSCs, the combination of simvastatin and marrow stem cells transplantation for the treatment of steroid-associated osteonecrosis of femoral head have good application prospects. PMID- 18396705 TI - [CT/MRI image characteristics of iliopsoas bursitis in avascular necrosis of femoral head]. AB - OBJECTIVE: To investigate the spectrum of CT and MR imaging and surgical operation findings in iliopsoas bursitis in patients with avascular necrosis of femoral head so as to enhance the diagnostic ability. METHODS: A total of 1,415 patients with avascular necrosis of the femoral head were analyzed retrospectively; of them, 15 patients were complicated by iliopsoas bursitis surgically or aspiration of synovial fluid between May 2005 and May 2007. Fifteen cases were all necrosis of the bilateral femoral head and 17 hips were combined with iliopsoas bursitis. There were 14 males and 1 female, aging 29-58 years. The course of disease was 1 month to 3 years. All 15 patients had limitation of ability of the hips and the "4" type sign was positive. The Harris score of hip's function was 54-78 (mean 62.7). Five patients of them can be touched a palpable cystic mass and tenderness in the inguinal area, and 3 of them associated with femoral neuropathy and 2 patients presented slight atrophy of the thigh muscle in suffering side. All these cases were taken X-ray films of positive and frog-leg lateral position, helical CT scan with 5 mm thinness, and MRI was performed in 6 patients with TlWI, T2WI, T2WI and fat-saturated inversion recovery sequence. RESULTS: The radiographs were the primary basis evidences for diagnosis and degrees of the avascular necrosis of femoral head. According to the standards of Association Research Circulation Osseuse, there were 2 hips at stage II (II C 2), 6 hips at stage III ( II B 1, III C 5 and 9 hips at stage IV. The X-ray films showed the bulging of the fat pad and soft tissue swelling in 6 patients. CT analysis disclosed that the enlarged iliopsoas bursae appeared as hypodense, well defined, thin-walled (< 2 mm) cystic structures. The content of the examined bursae was homogeneous with a CT density of ranging from 12.7 to 41.2 Hu, showing fluid collection. They were round or oval in shape medial to the iliopsoas, exhibiting inyvrted water-drop cystic shadow just inferior to the femoral head. Slight contrast enhancement of the bursal wall was seen after contrast agent administration in 3 cases. MRI demonstrated that the iliopsoas bursitis presented as low signal on T1WI and water-like high signal on T2WI and markedly higher signal on STIR in 6 cases. The demonstration of the extent, size, mass effects and its relation and subsequent affection to surrounding anatomical structures were clearly shown by MRI, and by the communications between the il opsoas bursa and the adjacent hip joint. CONCLUSION: In the diagnosis of avascular necrosis of femoral head with imaging approaches, much attention should be paid to the abnormalities around the articular capsule to early identify iliopsoas bursitis for further management. PMID- 18396706 TI - [Study on correlation between pain grading, stage of necrosis and bone marrow edema in nontraumatic osteonecrosis of femoral head]. AB - OBJECTIVE: To explore the correlation between pain grading, stage of necrosis and bone marrow edema (BME) in nontraumatic osteonecrosis of femoral head (NONFH) so as to strengthen understandings about clinical significance of BME in NONFH. METHODS: From October 2004 to October 2006, 97 patients (149 hips) with NONFH were treated. There were 68 males and 29 femals with an average age of 38.8 years (19-62 years). The disease course was from 20 days to 4 years. BME was identified grade 0 to grade 2 according to MRI. Based on grading scale of pain, pain grading were divided into no pain (grade 0), mild pain (grade 1) and moderate or severe pain (grade 2). According to Association Research Circulation Osseous staging system, NONFH were divided into I-IV stages. The incidence rate of BME in each pain grading and stages of necrosis was analyzed respectively. Contingency table analyses and rank sum tests were used to compare the difference of pain grading and stages of necrosis among these groups. RESULTS: The total incidence rate of BME was 73.15% (109/149), the incidence rates were 84.38% in pain groups (108/128) and 94.12% in the grade 2 (32/34). Pain grading correlated with BME rating (P < 0.001). The results of rank sum tests for several independent samples showed significant difference in BME among pain groups(P < 0.001). With the advance of pain scale, the mean rank of BME increased gradually (28.19 for grade 0, 78.94 for grade 1 and 96.12 for grade 2). BME was more commonly and clearly seen in stage II (77.05%) and stage III (82.81%) of NONFH. Stage I-III of NONFH correlated with BME rating (P < 0.001). The results of rank sum tests showed significant difference in BME rating among three stages (P < 0.001). With the advance of disease, the rank of BME rating increased gradually (39.07 for grade 0, 60.16 for grade 1 and 86.15 for grade 2). CONCLUSION: BME is a sign that is accompanied with NONFH. The probability and extent of BME correlated well with the pain and stage of NONFH. The condition of BME can be used as a index for the appraisal of advancement of disease and the judgment of treatment result. PMID- 18396707 TI - [Mechanism of "crescent sign" formation in avascular necrosis of femoral head]. AB - OBJECTIVE: To investigate corresponding relation between structure change of the femoral head with "crescent sign" and stress exerted on the avascular necrosis of femoral head, to explore the mechanism of the "crescent sign" formation. METHODS: From March 1998 to April 2003, the femoral heads of 18 hips in 16 cases having osteonecrosis and "crescent sign" in X-ray film before total hip arthroplasty, were collected. General and coronal section plane morphology of the femoral heads were observed. The principle of effective stress and stress concentration theory were used to explain the phenomena and structure changes in osteonecrosis of the femoral head. RESULTS: Cancellous bone existed as a three-dimensional, interconnected network of trabeculae rods and plates, with 50%-90% of porosity and 20-30 mmHg bone marrow pressure. According to the definition of porous media, bones especially cancellous bone was a kind of solid and liquid two phases porous media. Cross-sectional structure changes in the junction between subchondral plate and cancellous were the place where stress concentrated. The principle of effective stress and stress concentration theory could explain the phenomena and their relationship that occurred in avascular necrosis of the femoral head. CONCLUSION: The "crescent sign" starts in an area of very focal resorption in the subchondral plate laterally and peripherally. The focal resorption in the subchondral plate breaks the continuity of subchondral plate and causes stress concentration in the resorption region. The concentrated stress accumulates in the junction between subchondral plate and unrepaired necrotic cancellous bone brings on the fracture right below the subchondral plate. The focal resorption of the subchondral plate also provides a pathway for the pore water in the unrepaired necrotic bone skeleton to outflow, therefore cause effective stress increase and unrepaired necrotic bone skeleton be compacted by increased effective stress applied on unrepaired necrotic cancellous bone skeleton, and results in the volume decrease of unrepaired necrotic cancellous bone and the formation of cavum below the subchondral plate. The cavum shows "crescent sign" in the X-ray film. PMID- 18396708 TI - [Implantation of calcium phosphate cement/Danshen drug delivery system for avascular necrosis of femoral head]. AB - OBJECTIVE: To introduce a new method using calcium phosphate cement/Danshen drug delivery system for avascular necrosis of femoral head and to evaluate its clinical outcome. METHODS: From May 2000 to June 2005, 48 patients (54 hips) with avascular necrosis of femoral head were treated with calcium phosphate cement/Danshen drug delivery system implantation in the involved femoral head. There were 32 males (36 hips) and 16 females (18 hips) with an average age of 38.7 years (26-62 years). Twenty-one cases had the history of drinking or smoking, 15 cases had the history of receivihg hormonotherapy and 2 had the history of injury in hip joint. The disease course was 2-32 months. According to standard of Association Research Circulation Osseous (ARCO) staging, 9 hips were classified as stage I, 31 as stage II and 14 as stage III The operation consisted of removal of necrotic bone under weight-loading cartilage and the implantation of calcium phosphate cement/Danshen drug delivery system, all manipulations were done through a bone tunnel in trochanter. The function of hip joint were evaluated and X-ray films were taken pre- and post-operatively. RESULTS: No phlebothrombosis of leg and foreign body action occurred in all cases, and incision healed by first intention. The postoperative follow-up averaged 42.5 months, ranging from 22 to 73 months. According to the evaluation criterion of Dandong 1995 for adult avascular necrosis of femoral head, the results were excellent in 33 hips, good in 17, fair in 3 and poor in 1, the excellent and good rate was 92.6%. CONCLUSION: This method is relatively simple with less invasion, it not only improves the microcirculation of femoral head by local application of traditional Chinese medicine, but also provide mechanic buttress in the weight loaded area, which is beneficial to repair and reconstruction of femoral head. It may be a choice of minimally invasion surgery for femoral head necrosis. PMID- 18396709 TI - [A prospective study on wound-healing hydrogel in treating chronic venous ulcer of lower extremities]. AB - OBJECTIVE: To study the outcome of wound-healing hydrogel in treating chronic venous ulcer of lower extremities so as to find a new therapy. METHODS: From April 2007 to September 2007, 60 patients with chronic venous ulcer of lower extremities were randomly assigned to wound-healing hydrogel group (group A, 30 cases) and control group (normal saline, group B, 30 cases). In group A, there were 24 males and 6 females, aging (57.3 +/- 6.8) years; the disease course was (2.9 +/- 0.7) years; and the ulcer area was (3.4 +/- 0.6) cm2. In group B, there were 20 males and 10 females, aging (60.1 +/- 7.4) years; the disease course was (3.3 +/- 0.9) years; and the ulcer area was (3.1 +/- 0.4) cm2. There were no differences in age, area of ulcer and course of disease between two groups (P > 0.05). The area of ulcer was measured every week after the treatment, and the effect of treatment was evaluated after 15 days. RESULTS: The ulcer area of 7 days and 14 days after treatment was (2.6 +/- 0.7) and (1.1 +/- 0.2) cm2 in group A, and (2.8 +/- 0.6) and (2.3 +/- 0.7) cm2 in group B, respectively; showing no statistically significant differences 7 days after treatment (P > 0.05), and showing statistically significant difference 14 days after treatment between two groups (P < 0.05). The average healing time was (12.0 +/- 1.7) days in group A, and (31.0 +/- 2.9) days in group B, respectively, showing statistically significant difference (P < 0.01). The results were excellent, good, fair and poor in 16, 9, 4 and 1 of group A, and were in 3, 9, 14 and 4 of group B, respectively; showing statistically significant difference (P < 0.01). CONCLUSION: Wound-healing hydrogel is effective in treating chronic venous ulcer of lower extremities. PMID- 18396710 TI - [Less invasive leverage reduction with external fixator supported and bone graft for treatment of unstable fractures of distal radius]. AB - OBJECTIVE: To assess the results of treatment of unstable distal radius fractures with leverage reduction and bone graft assisted by external fixators. METHODS: From September 2005 to May 2007, 27 cases of unstable distal radius fractures were treated by leverage reduction and bone graft, meanwhile assisted by external fixators. The cases included 16 males and 11 females, aged from 18 to 69 years with an average of 49.3 years. Fractures were caused by falling in 19 cases, crash from high place in 1 case, traffic accident in 6 cases, and obtuse strike directly in 1 case, which were all closed fresh bone fractures. According to the standard of AO, all cases were classified as type C1 in 13 cases, type C2 in 11 cases, and type C3 in 3 cases. The palmar inclinination was from -38 degrees to 10 degrees (mean -12.2 degrees); the ulnar deviation angle was from 6 degrees to 30 degrees (mean 19.1 degrees) before operations. The operations were performed from 1 to 3 days after injuries. The function of the carpal joints and the bone healing conditions were evaluated after operations. RESULTS: All cases were followed up for 4-24 months (mean 13.1 months). No complications such as pin loosening dislocation of fixators, injury of blood vessels and radial nerves, pin track infections occurred. According to Mcbride scoring, the results were excellent in 10 cases, good in 13 cases, fair in 3 cases and poor in 1 case, the excellent and good rate being 85.2%. One case had traumatic arthritis and 1 case had wrist joint stiffness. All achieved fractures uniton 8-10 weeks (mean 9.3 weeks) after operations. The palmar inclinination angle was from 0 degree to 20 degrees (mean 13.40 degrees); the ulnar deviation angle was from 10 degrees to 33 degrees (mean 22.1 degrees) after operations. CONCLUSION: Treatment of unstable fractures of the distal radius by use of leverage reduction and bone graft with external fixator offers many advantages, such as simple operation, satisfactory reduction, rigid fixation, excellent function and lower incidence rate of traumatic arthritis. PMID- 18396711 TI - [Free vascularized fibular associated with iliac graft to treat old femoral neck fracture]. AB - OBJECTIVE: To evaluate the clinical outcome of free vascularized fibular associated with iliac graft in treatment of old femoral neck fracture. METHODS: From January 1994 to January 1997, 76 cases of old femoral neck fracture were treated with free vascularized fibular associated with iliac graft, including 54 males and 22 females, aging from 24 to 48 years with an average of 31.5 years. All of these fractures resulted from injury. There were 20 cases of Garden II, 41 of Garden III and 15 of Garden IV. Based on the location of fracture, there was 26 cases of subcapital, 42 cases of transcervical and 8 cases of basal. Sixty five cases were treated with internal fixation, 7 cases with skin traction and 4 just with staying in bed. Their Harris score were from 52 to 72 with an average of 65.6. The time from injury to operation was 2-24 months. The size of free vascularized fibular was 6-8 cm and iliac graft was 3.0 cm x 2.0 cm x 1.5 cm. RESULTS: In 76 cases, 68 were followed up and all fractures healed within 4 to 6 months with an average of 5.2 months. The increased density in femoral head was observed 1 year after operation. After 10 years of operation normal hip function was achieved in 63 cases followed up 10.1-12.4 years with the average as 10.5 years); the Harris score was 87.5 (84 to 94). The structure of femoral head was normal and the grafted fibular and iliac bone healed with the femoral, no elapse or cyst occurred. Five cases had been complied with total hip replacement for femoral head necrosis or other. CONCLUSION: Free vascularized fibular associated with iliac graft is a good method to treat old femoral neck fracture. PMID- 18396712 TI - [Evaluation of percutaneous iliosacral screws for unstable pelvic fractures]. AB - OBJECTIVE: To evaluate the effectiveness of percutaneous fixation with iliosacral screw in the management of unstable pelvic fractures. METHODS: From March 2003 to January 2007, 15 patients with unstable pelvic fractures were treated by percutaneous fixation with iliosacral screws. There were 6 males and 9 females, aged 21-56 years. Fractures were caused by traffic accident in 8 cases, high falling in 6 cases and crushing injury in 1 case. The disease course ranged from 4 hours to 3 days. Of 15 patients, 7 had fractures of pubium and sacrum; 2 had fractures of ilium and sacrum; 4 had dislocation of sacroiliac joint with fractures of pubic rami; and 2 had dislocation sacroiliac joint with fracture of sacrum of pubium. Anterior fixation was performed by means of plating in 3 cases, by external fixation in 7 cases, and by both in 5 cases. RESULTS: The blood loss was from 1,000 to 1,500 mL in 4 patients with acetabular fracture and about 50 mL in the others. The average operative time was 153.6 minutes (range, 60-305 minutes). Fifteen patients were followed up 6 months to 3 years with an average of 18 months. According to Matta criterion for fracture reduction, the results were excellent in 14 cases and good in 1 case. After a follow-up of 28.8 months, fracture union was achieved in all patients. Screw loosening occurred in 1 case, pain of lumbosacral region in 3 cases after tiredness and mild claudication in 2 cases. Nine patients returned to their occupation, three patients changed occupation, and 3 patients could not work at last follow-up. According to Majeed functional scoring, the results were excellent in 11 cases and good in 4 cases. CONCLUSION: Percutaneous iliosacral screw fixation is a useful method in treatment of unstable pelvic fracture. PMID- 18396713 TI - [Treatment of old Tile B1 pelvic fracture by staged pelvic closure using external fixator combined with reconstruction plate]. AB - OBJECTIVE: To summary the effects of staged pelvic closure using external fixator combined with reconstruction plate for old Tile B1 pelvic fracture. METHODS: From August 2000 to August 2006, 14 patients (9 males and 5 females, age ranging from 21 to 65 years with old Tile B1 pelvic fracture with pubic symphysis separation were treated. The injuries were caused by the traffic accident in 10 patients, high crash in 3, and crush in 1. The duration ranged from 4 weeks to 3 months. The 14 patients were complicated with other injuries in some degrees. The X-ray and CT showed pubic symphysis separation. In 13 patients, pubic symphysis separation distance was more than 2.5 cm, who also had fracture or dislocation in the posterior structure of pelvis. The X-ray films showed the mean pubic symphysis separation distance was (6.67 +/- 2.11) cm preoperatively. The 14 patients underwent pelvic external fixation at first and staged pelvic closure gradually for the pubic symphysis separation. After 2 to 3 weeks, when the pubic symphysis separation distance was less than 1 cm, the patients underwent open reduction and internal fixation with reconstruction plate. Pubic symphysis separation distance was measured in the preoperative and postoperative pelvic anterioposterior X-ray films. The condition of the posterior structure of pelvis was observed in CT films. The functions of patients were assessed according to Majeed grading system. RESULTS: All incisions healed by first intention. The 14 patients were followed up for 6 months to 2 years (15 months on average). The X ray films showed the mean pubic symphysis separation distance of post-operation was (0.85 +/- 0.23) cm, showing statistically significant difference when compared with that of pre-operation (P < 0.05). The CT films showed fracture and dislocation of the posterior structure of pelvis had bony healing after 6 to 12 months. According to the Majeed grading system, the results were excellent in 5 cases, good in 4 cases, fair in 4 cases and poor in 1 case; the excellent and good rate was 64.29%. CONCLUSION: Staged pelvic closure using external fixation combined with reconstruction plate for old pelvic fracture with pubic symphysis separation can reduce the pubic symphysis separation distance significantly. Satisfactory effects can be expected in treating the patients with Tile B1 fracture. PMID- 18396714 TI - [Facial and cervical reconstractions using expanded flap from medial upper arm]. AB - OBJECTIVE: To investigate the blood supply of the expanded skin flap from the medial upper arm and its application for the repair of facial and cervical scar. METHODS: From May 2000 to February 2007, 20 cases (12 males and 8 females; aging from 7 to 42 years) of facial and cervical scar were treated with the expender flap from medial upper arm. The disease course was 9 months to 20 years. The size of the scar was 8 cm x 6 cm-22 cm x 18 cm. The operation was carried out for three steps: (1) The expander was embed under the superior proper fascia. (2) The scar in the face and cervix was loosed and dissected. Combined the expanded skin flap from the medial upper arm (the size of the flap was 9 cm x 7 cm-24 cm x 18 cm) in which the blood supply to the flap was the superior collateral artery and the attributive branches of the basilica with auxiliary veins for blood collection with partial scar flap (3.5 cm x 2.5 cm-8.0 cm x 6.0 cm) was harvested and transferred onto the facial and cervical defect. (3) After being cut off the pedicle, the scar was dissected. The expanded flap was employed to cover the defect. RESULTS: After 3-24 months follow-up with 16 cases, all the grafted skin flaps survived at least with nearly normal skin color, texture and contour. The scars at the donor sites were acceptable. The function and appearance of the face and cervix was improved significantly. No surgery-related significant complications were observed. CONCLUSION: Repair of facial and cervical scar with the medial upper arm expanded skin flap is a plausible reconstructive option for head and face reconstructions. However, a longer surgery time and some restrictive motion of the harvested upper limbs might be a disadvantage. PMID- 18396715 TI - [Application of residual ear in auricular reconstruction of microtia]. AB - OBJECTIVE: To summarize different treatments of the residual ear in auricular reconstruction, to investigate the reasonable applications of the residual ear. METHODS: From September 2005 to July 2006, 128 patients (79 males, 49 females; aging 5-21 years with an average of 11 years) with unilateral microtia underwent the staged repair. In the patients, there were 44 cases of left-unilaterally microtia and 84 cases of right-unilaterally microtia. The residual ears looked like peanut in 56 patients, like sausage in 35 patients, like boat in 27 patients, and like shells in 10 patients. Among all the patients, the external acoustic meatus was normal in 5 patients, stenosis in 11 patients, and atresia in 112 patients. According to auricular developmental condition, the patients were divided into three types: 17 cases of type I, 98 cases of type II, and 13 cases of type III. In the first stage operation, a 50 mL kidney-liked expander was implanted into post aurem subcutaneous tissue. The residual ear whose superior extremity was close to the hair line was treated. The middle and superior part of the residual ear was cut. The redundant residual auricular cartilage was removed. In the second stage operation, the inferior part of the cartilage frame was covered by the middle and superior part of the residual ear. According to the location of the residual ear, "V-Y" plasty, "Z"-plasty and reversal of the residual ear were used to correct the location of the residual ear. In the third stage operation, the remained residual ear was used to reconstruct crus of helix or cover the wound surface which was resulted from repairing the reconstructed ear. RESULTS: The residual ears which were reshaped and transferred had good blood circulation. All residual ears were survival. The wounds healed by first intention. The follow-up for 8-15 months showed that the auricular lobule of the reconstructed ear was turgor vital is and natural. The locations of the reconstructed ear and normal side ear were symmetry. The auricular lobules of the reconstructed ear survived well. The reconstructed crus of helix, helix, antihelix and triangular fossa were clear. The results were satisfactory. CONCLUSION: Using residual ear reasonably is an important procedure of successful auricular reconstruction and the symmetry of the reconstructed ear and uninjured side ear. PMID- 18396716 TI - [Clinical application of decalcified dentinal matrix of human to repair alveolar cleft]. AB - OBJECTIVE: To study a new method of alveolar cleft bone graft repair. METHODS: From April 2005 to August 2006, 26 cases (30 sides) of alveolar cleft were treated with trabecular substance of self-ilium combined with the decalcified dentinal matrix of human (DDM). There were 16 males and 10 females, aged 6-12 years. Unilateral alveolar cleft was involved in 22 cases, bilateral alveolar cleft in 4 cases. All cases were accompanied by nasal wing collapse, 5 by deciduous tooth retention, 3 by malposed teeth and 1 by tooth deformity. Anterior occlusal radiographs and panoramic oral radiographs were taken to observe union and bone absorption before and after operation. RESULTS: Twenty-three patients achieved healing by first intention, oronasal fistulas were closed successfully. Infection occurred in one bilateral alveolar cleft case and bone tissue exposure in 2 cases. Wound healed by rinse or dressing change and inunction with MEBO. The X-ray films after operation showed bone bridge formation in the alveolar cleft. The stability and continuity of body of maxilla were resumed. Four cases (6-9 years old) were observed 6.5 months, 22 cases (9-12 years old) were observed 8.6 months. According to Bergland for evaluation, the X-ray films after 3 months of operation showed 16 cases (16 sides) in class I (53.3%), 7 cases (8 sides) in class II (26.7%), 2 cases (4 sides) in class III (13.3%), and 1 case (2 sides) in class IV (6.7%). The overall survival ratio of alveolar bone grafting was 93.3%, and the clinical success ratio was 80%. CONCLUSION: The DDM is good in alveolar cleft bone graft repair. It is better than using self-ilium only clinically. PMID- 18396717 TI - [Autologous free fat particle grafting combined with bFGF to repair facial depression]. AB - OBJECTIVE: To evaluate the effect of autologous free fat particle grafting combined with-bFGF to repair facial depression. METHODS: From April 2004 to May 2006, 41 patients with facial depression were randomized into two groups (groups A and B). In group A, 12 cases were admitted from April 2004 to December 2004. There were 5 males and 7 females, aging 16-49 years (mean, 31 years). The pathological causes were congenital facial depression in 2 patients, hemifacial atrophy in 2, traumatic cicatrix in 5 and benign tumor removal in 3. The course of disease was 2-19 years. The concave regions were low (0.52 +/- 0.13) cm compared to surrounding normal skin, concave area (16.0 +/- 5.3) cm2. In group B, 29 cases were admitted from January 2005 to May 2006. There were 14 males and 15 females, aging 18-52 years (mean, 37 years). The pathological causes were: congenital facial depression in 3 patients, hemifacial atrophy in 4, traumatic cicatrix in 15 and benign tumor removal in 7. The course of disease was 2-20 years. The concave regions were low (0.58 +/- 0.15) cm compared to surrounding normal skin, concave area (18.0 +/- 6.2) cm2. Cases in group A were treated with pure autologous free fat particle injection; cases in group B were treated with autologous free fat particle injection combined with bFGF (4,200 IU/10 mL). The clinical outcome were comparatively analyzed between two groups after operation. RESULTS: The follow-up time was 6 to 24 months (mean, 12.5 months) in group A and 6 to 24 months (mean, 13 months) in group B. In group A, 6 patients achieved satisfactory clinical effect after one injection of fat particle, the satisfactory rate of one therapy being 50%; other 6 cases were required reinjection of fat particle 6-12 months postoperatively, of which two-time injections in 3 cases, three-time injections in 3 cases. In group B, 24 patients achieved satisfactory clinical effect after one injection of fat particle, the satisfactory rate of one therapy being 82.8%; only 5 cases were required reinjection one year postoperatively. There was statistically significant difference in the satisfactory rate of one injection between two groups (P < 0.05). CONCLUSION: Autologous free fat particle grafting combined with bFGF to treat facial depression can acquire satisfactory clinical effect, which is a safe and effective method. PMID- 18396718 TI - [Clinical evaluation of effect of transplantation by auto-fat granule injection for mastatrophy post suckling]. AB - OBJECTIVE: To investigate the clinical effect of transplanting by auto-fat granule injection for mastatrophy post suckling. METHODS: From March 2000 to June 2006, 73 patients (146 breasts) with mastatrophy post suckling were treated by transplanting auto-fat granule. The mastatrophy occurred between ages 28 and 52 years with a median of 37 years post suckling. The breasts shrank and their elasticity decreased gradually within 2-10 years post suckling. The auto-fat granule was obtained by liposuction with syringe from patient's abdomen, waist, buttocks and thighs, etc. After repeated wash and purification, the auto-fat granule was transplanted into the interspace behind the breast by injection. The quantity of auto-fat granule was 50-100 mL in each side of breast per transplantation at 3-6 months intervals, and the whole course of treatment needed 2-6 transplantations. RESULTS: The incisions in all cases healed primarily postoperatively. In 73 cases, 65 were followed up from 6 months to 3 years post operation. All patients had a significant improvement in their breast size and shape postoperatively and their breasts were soft and natural in appearance and feel. All of them had more perfect arcuation of physique and body with strengthened self-confidence, relieved mood and improved quality of life. However, small indurations were found sporadically in 7 cases (10 breasts) within 2-7 months, and calcifications in 5 cases (8 breasts) within 9-14 months post the first operation. CONCLUSION: The transplantation by auto-fat granule injection for mastatrophy post suckling is an effective and practical method. The surgical technique is well worth performing in clinical practice. PMID- 18396719 TI - [Analysis of reasons of tendon adhesion post tendon allograft]. AB - OBJECTIVE: To explore the reasons of tendon adhesions post tendon allograft. METHODS: From May 1990 to June 2000, 85 cases receiving tendon allograft were given tenolysis because of tendon adhesions. There were 76 males and 9 females, with an average age of 24.5 years (8-46 years). Injury was caused by machine in 38 cases, electric in 32 cases, cut in 4 cases, explosion in 4 cases and extremity mutilation in 7 cases; including 66 cases of flexor tendon deficit and 19 cases of extensor tendon deficit. Six cases had 1 tendon deficit, 79 cases had tendon deficit of more than 2. The defect region ranged from I to V. The total mobility of the joint was less than 220 degrees in 73 cases. The impairment of skin, bone, nerve and vascular were treated before tendon allograft. RESULTS: Because TAM was less than 50% of TPM, the patients were given tenolysis 4-15 months after operation. And the mobilization began at the first day after operation to improve the range of active movement. Patients were followed up 7-17 years (mean 12.7 years). TAM and TPM were in accord. Mean total mobility of joint was 200 degrees. CONCLUSION: The serious of primary hurt is the important factors of tendon adhesion. Improvement of tendon selected, treatment and early mobilization can relieve the tendon adhesion. PMID- 18396720 TI - [Preventing effect of TGF-beta1 antibody compounded with fibrin glue on postoperative adhesions of flexor tendon]. AB - OBJECTIVE: To explore the preventing effects of TGF-beta1 antibody (TGF-beta1Ab) compounded with fibrin glue (FG) on postoperative adhesions of flexor tendon. METHODS: Seventy-two Leghorn chickens were randomly divided into 4 groups (groups A, B, C and D), 18 chickens for each group, and the long flexor tendons of the 3rd and 4th toes in zone II of all chickens were transversed and sutured with the 4-strand cruciate repair technique to make defect models. In group A, 0.2 mL TGF beta1 Ab was applied at repair site. In group B, 0.2 mL FG was applied at repair site. In group C, 0.2 mL TGF-beta1Ab and FG was applied at repair site. In group D, 0.2 mL normal sodium was applied at repair site. At 1, 3 and 8 weeks after operation, the tendons of 6 chickens in each group were harvested for morphological and histological evaluation. Six specimens of each group were obtained for biomechanical test at 3 and 8 weeks. RESULTS: The gross-observation showed that the differences in grading of tendon adhesion were not significant among 4 groups at 1 week after operation (P > 0.05), but the differences were significant between groups A, B, D and group C at 3 and 8 weeks after operation (P < 0.05). Histological observation showed that collagen fibers arranged irregularly in groups A, B and D, but arranged regularly in group C at 3 and 8 weeks' after operation. At 3 weeks after operation the gliding excursion ratio of the tendon in groups A, B, C and D were 0.45 +/- 0.05, 0.40 +/- 0.10, 0.79 +/- 0.09 and 0.25 +/- 0.07 respectively; the simulated active flexion ratio were 0.61 +/- 0.02, 0.67 +/- 0.03, 0.91 +/- 0.03 and 0.53 +/- 0.04 respectively; the work of flexion were (18.00 +/- 0.77), (17.80 +/- 1.13), (27.60 +/- 1.73) and (15.60 +/- 1.27) degrees/N respectively. There were significant differences between group C and other three groups (P < 0.05). The tendon anastomosis breaking strength were (14.2 +/- 1.9), (15.2 +/- 2.2), (16.0 +/- 2.2) and (14.7 +/- 2.7) N, showing no significant differences among 4 groups (P > 0.05). At 8 weeks after operation, the gliding excursion ratio of the tendon in groups A, B, C and D were 0.45 +/- 0.07, 0.43 +/- 0.08, 0.80 +/- 0.09 and 0.29 +/- 0.05 respectively; the simulated active flexion ratio were 0.61 +/- 0.02, 0.63 +/- 0.03, 0.92 +/- 0.03 and 0.53 +/- 0.03 respectively, the work of flexion were (18.30 +/- 0.84), (18.60 +/- 0.80), (27.90 +/- 1.24) and (15.30 +/- 0.75) degrees/N respectively. There were significant differences between group C and other three groups (P < 0.05). The tendon anastomosis breaking strength were(51.9 +/- 3.0), (51.4 +/- 1.4), (53.3 +/- 1.3) and (52.3 +/- 2.2) N, showing no significant differences among 4 groups (P > 0.05). CONCLUSION: TGF-beta1Ab compounded with FG could significantly prohibit the formation of fibrous adhesions without interfering with the healing process. PMID- 18396721 TI - [Influence of human tissue factor pathway inhibitor gene transfection on neointima formation in vein grafts]. AB - OBJECTIVE: To reduce restenosis in vein grafts after coronary artery bypass grafting, to investigate the effect of human tissue factor pathway inhibitor (TFPI) gene delivery on neointima formation. METHODS: The eukaryotic expressed plasmid vector pCMV-(Kozak) TFPI was constructed. Forty-eight Japanese white rabbits were randomly divided into 3 groups with 16 rabbits in each group: TFPI group, empty plasmid control group and empty control group. Animal model of common carotid artery bypass grafting was constructed. Before anastomosis, vein endotheliocytes were transfected with cationic liposome containing the plasmid pCMV-(Kozak) TFPI (400 microg) by pressurizing infusion (30 min) in TFPI group. In empty plasmid control group, vector pCMV-(Kozak) TFPI was replaced by empty plasmid pCMV (400 microg). In empty control group, those endotheliocytes were not interfered. After operation, vein grafts were harvested at 3 days for immunohistochemical, RT-PCR and Western-blot analyses of exogenous gene expression and at 30 days for histopathology measurement of intimal areas, media areas and calculation of intimal/media areas ratio. Luminal diameter and vessel wall thickness were also measured by vessel Doppler ultrasonography and cellular category of neointima was analyzed by transmission electron microscope at 30 days after operation. RESULTS: Human TFPI mRNA and protein were detected in TFPI group. The mean luminal diameter of the TFPI group, empty plasmid control group and empty control group was (2.68 +/- 0.32) mm, (2.41 +/- 0.23) mm and (2.38 +/- 0.21) mm respectively. There were statistically significant differences between TFPI group and control groups (P < 0.05). The vessel wall thickness of the TFPI group, empty plasmid control group and empty control group was (1.09 +/- 0.11) mm, (1.28 +/- 0.16) mm and (1.34 +/- 0.14) mm respectively. There were statistically significant differences between TFPI group and other control groups (P < 0.01). The mean intimal areas, the ratio of the intimal/media areas of the TFPI group were (0.62 +/- 0.05) mm2 and 0.51 +/- 0.08 respectively,whichwere reduced compared withthose of the two control groups (P < 0.05). The mean media areas had no significant differences among three groups (P > 0.05). Through transmission electron microscope analyses,no smooth muscle cells were seen in neointima of TFPI group in many visual fields,but smooth muscle cells were found in neointima of two control groups. CONCLUSION: Human TFPI gene transfection reduced intimal thickness in vein grafts. PMID- 18396722 TI - [Fabrication of a novel cartilage acellular matrix scaffold for cartilage tissue engineering]. AB - OBJECTIVE: To develop a novel cartilage acellular matrix (CACM) scaffold and to investigate its performance for cartilage tissue engineering. METHODS: Human cartilage microfilaments about 100 nm-5 microm were prepared after pulverization and gradient centrifugation and made into 3% suspension after acellularization treatment. After placing the suspension into moulds, 3-D porous CACM scaffolds were fabricated using a simple freeze-drying method. The scaffolds were cross linked by exposure to ultraviolet radiation and immersion in a carbodiimide solution 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N hydroxysucinimide. The scaffolds were investigated by histological staining, SEM observation and porosity measurement, water absorptiofl rate analysis. MTT test was also done to assess cytotoxicity of the scaffolds. After induced by conditioned medium including TGF-beta1, canine BMSCs were seeded into the scaffold. Cell proliferation and differentiation were analyzed using inverted microscope and SEM. RESULTS: The histological staining showed that there are no chondrocyte fragments in the scaffolds and that toluidine blue, safranin O and anti-collagen II immunohistochemistry staining were positive. The novel 3-D porous CACM scaffold had good pore interconnectivity with pore diameter (155 +/- 34) microm, 91.3% +/- 2.0% porosity and 2451% +/- 155% water absorption rate. The intrinsic cytotoxicity assessment of novel scaffolds using MTT test showed that the scaffolds had no cytotoxic effect on BMSCs. Inverted microscope showed that most of the cells attached to the scaffold. SEM micrographs indicated that cells covered the scaffolds uniformly and majority of the cells showed the round or elliptic morphology with much matrix secretion. CONCLUSION: The 3-D porous CACM scaffold reserved most of extracellular matrix after thoroughly decellularization, has good pore diameter and porosity, non-toxicity and good biocompatibility, which make it a suitable candidate as an alternative cell carrier for cartilage tissue engineering. PMID- 18396723 TI - [Simplified preparation and relative evaluation of decellularized porcine aortic scaffold]. AB - OBJECTIVE: To investigate the feasibility of applying enzymatic method to prepare decellularized porcine aorta and to evaluate its biomechanical properties, immunogenicity and cell compatibility. METHODS: 0.1% trypsin-0.01% EDTA was applied to extract cells from porcine aorta under 37 degrees C continuously vibrating condition and its histology and microstructure were observed. The thickness, stress-strain curve, ultimate tension stress (UTS) and strain of failure (SOF) were compared before and after decellularization for 48, 96 and 120 hours under uniaxial tensile tests, respectively. The histological change was observed at 1, 3 and 6 weeks after the decellularized tissue was implanted subcutaneously in 3 dogs. According to the HE stains and a semi-quantitative Wakitani grading method, gross changes, category and amounts of infiltrated cells and neo-capillaries were compared between pre- and post-decellularization of porcine aortae. Endothelial cells from canine external jugular vein were seeded onto the decellularized patches to observe the cell compatibility. RESULTS: Microscopy and electron microscopies examination identified that cell components was completely removed from the fresh porcine aorta and Masson's trichrome showed that the structure of matrix (fibrins) was maintained intact at 96 hours using the decellularization method. There were no significant differences in the thickness, UTS and SOF between before and after decellularization (P > 0.05). However, The UTS values showed a decrease tendency and SOF showed an increase tendency. The stress-strain curve also verified a decrease tendency in mechanical intensity and an increase one in ductility after decellularization. After implanting the acellularized matrix subcutaneously in canine, moderately lymphocyte infiltration was seen at the 1st week and the infiltration was replaced by fibroblasts accompanied by neocapillary formation at the 6th week. A semi-quantity histological evaluation showed that there were differences in gross observation, category and the numbers of the infiltrated cells between decellularized and non-decellularized tissues (P < 0.05). A cell monolayer was identified by HE staining and scanning electron microscopy when the endothelial cells were seeded onto the inner luminal surface of the scaffold, aligned at the same direction on the whole. CONCLUSION: The decellularized porcine aortic scaffold, prepared by trypsin-EDTA extraction under continuously vibrating condition, could meet the requirements of tissue-engineering graft in biomechanical properties, immunogenicity and cell compatibility. PMID- 18396724 TI - [Constructing a small-diameter decellularized vascular graft pre-loaded with bFGF]. AB - OBJECTIVE: To design a novel small-caliber vascular graft using a decellularized allogeneic vascular scaffold pre-loaded with bFGF. METHODS: The decellularized canine common carotid were obtained by a detergent-enzymatic procedure, then the scaffolds were covalently linked with heparin and pre-loaded with bFGF, the amount of binding bFGF and releasing curve were assayed by ELISA. Canine BMSCs expanded in vitro were seed on the scaffolds to observe the effects of binding bFGF on proliferation. Both bFGF pre-loaded and non-pre-loaded decellularized grafts were implanted in canines as carotid artery interposition for 8 weeks, the patency was examined by digital subtraction angiography and histological method. RESULTS: Histology and electron microscopic examination of the decellularized scaffolds showed that cellular components were removed completely and that the extracellular matrix structure remained intact. The amount of binding bFGF positively related to the concentration of bFGF. There was a significant difference in the amount of binding bFGF between two different scaffolds throughout all bFGF concentrations (P < 0.05), and up to 100 ng/mL, the local and sustained release of bFGF from the heparin treated scaffolds were assayed up to 20 days. Additionally, MTT test showed the bFGF-preloaded scaffolds significantly enhanced the proliferation of seeded BMSCs in vitro compared with non-bFGF preloaded scaffolds at 3 days after seeding and thereafter (P < 0.01). Furthermore, in vivo canine experiments revealed that all 8 bFGF-pre-loaded scaffolds remained patent after 8 weeks of implantation, and host cell lined the lumen and populated the wall. Only 1 non-bFGF-pre-loaded scaffold was patent, and the other 7 grafts were occluded because of thrombsus formation. CONCLUSION: This study provides a new strategy to develop a small diameter vascular graft with excellent biocompatibility and high patency rate. PMID- 18396725 TI - Premature ejaculation: different pathophysiologies and etiologies determine its treatment. AB - Premature ejaculation (PE) is a frequent male sexual complaint. Its occurrence does not automatically imply the existence of a male sexual disorder. The current DSM definition of PE has a low-positive predictive value with a high associated risk for false-positive diagnoses of PE. A new classification in four well defined PE syndromes has recently been proposed for the pending DSM-V. According to this new classification there are different pathophysiologies, etiologies, and treatments of PE dependent on the underlying PE syndrome. Some types are particularly neurobiologically or medically determined and need drug treatment; other types, which are mainly psychologically determined, need psychotherapy or both. Moreover, according to the proposed new classification it is likely that only a small number of men with complaints of PE suffer from a neurobiologically determined PE disturbance. PMID- 18396726 TI - Are post-menopausal women "half-a-man"?: sexual beliefs, attitudes and concerns among midlife Chinese women. AB - The objective of this study was to explore the concerns and issues specific to sexual behaviors and interests among postmenopausal women in a metropolitan urban city in Southern China. This was a qualitative study using semi-structured focus group discussions (FGDs). Twenty-two informants aged 50 and above recruited through an ongoing research project on early post-menopausal women under the purview of the School of Public Health at the Chinese University of Hong Kong. The main outcome measures were themes identified as health concerns towards sex among postmenopausal women. Contrary to prior expectations and stereotypes, we found that the respondents were very open in discussing their sexual concerns and shared extensively from their personal experiences. Our respondents perceived significant differences in sexual needs between older men and women. Many respondents identified the effects of menopause and general health problems, as well as environmental limitations and communication problems between spouses as barrier to healthy sexual relationships within their marriages. Conversely, sexual disharmony was seen as an important cause of marital strife among older couples. There is a growing need in studying health concerns among the fast growing, Chinese middle-aged and older population. Our qualitative study revealed a wide gap in the knowledge regarding general and sexual health issues among post menopausal Chinese women. Raising awareness among health professionals of these issues is essential in addressing health concerns regarding this traditionally taboo subject within the Chinese context. PMID- 18396727 TI - Discrepancies between sexual desire and sexual activity: gender differences and associations with relationship satisfaction. AB - The nature of sexual satisfaction and its relationship with relationship satisfaction was examined in a population-based Finnish sample of men and women between 33 and 43 years. Both men and women wished to experience kissing and petting, sexual fantasies, oral sex, and vaginal intercourse more often. Sexual satisfaction, defined as no discrepancy between desired frequency and actual frequency of sexual behaviors, was associated with relationship satisfaction. In both sexes, sexual satisfaction with vaginal intercourse as well as kissing and petting was positively associated with relationship satisfaction, whereas higher desired and actual frequency of masturbation were negatively associated with relationship satisfaction. PMID- 18396728 TI - Assessing unwanted early sexual experiences in a South-African sample. AB - This study assessed the prevalence of unwanted early sexual experiences of 1434 first-year university students, of whom 409 (28.5%) reported exposure to early unwanted sexual experiences. Most respondents reported knowing the perpetrator (80.2%) and being exposed to the unwanted behavior once (175; 45.7%). The most common coercive practice was "talk" (109; 38%) followed by being scared because they were bigger and stronger (61; 21.3%). PMID- 18396729 TI - Not all affairs are created equal: emotional involvement with an extradyadic partner. AB - The goal of the current study was to explore how participants' recollection of their level of emotional involvement with an extradyadic partner related to the precipitants and outcomes of that involvement. Three hundred and forty-five undergraduates and 115 community individuals with a history of extradyadic involvement completed self-report measures about their most recent extradyadic experience. In both samples, greater emotional involvement with an extradyadic partner was related to dissatisfaction with the intimacy in the primary relationship prior to the affair, intimacy and self-esteem motivations for involvement, gradual onset of attraction to the extradyadic partner, lower remorse regarding the involvement, a greater likelihood of primary partner knowledge of the involvement, and (in the community sample) marital dissolution. A path model further identified interrelationships among the variables, such as direct effects of dissatisfaction with intimacy in the primary relationship and concern for the primary partner's feelings. Emotionally involved affairs are different from casual affairs in terms of precipitants, motivations, development, and outcomes, with implications for prevention, intervention, and understanding the dangers of different types of affairs. PMID- 18396731 TI - Part III: tradition in transition. Can technology bridge the personnel gap? PMID- 18396730 TI - Psychotherapists' experience with clients who engage in consensual sadomasochism: a qualitative study. AB - Consensual sadomasochism (Bondage and Discipline, Dominance and Submission, Sadism and Masochism; BDSM) is relatively common, but the experience of psychotherapists who work with clients who engage in BDSM has received little study. We conducted semistructured interviews with 14 therapists experienced in working with BDSM clients. Interviews were recorded, transcribed, and analyzed for thematic content. Therapists emphasized the importance of cultural competence, including a nonjudgmental attitude and knowledge of BDSM practices and cultural values. BDSM was rarely a central issue in therapy, relationship issues were clients' most common presenting concerns. Therapists who practiced BDSM themselves often encountered boundary issues with clients. PMID- 18396732 TI - Measuring productivity. PMID- 18396733 TI - IBS vs. IBD and other tummy aches. PMID- 18396734 TI - Hitting the target with antithrombotic therapy. AB - Thrombus treatment and prevention can be regulated by a number of intravenous or subcutaneous drugs, as well as oral warfarin. Many therapies require laboratory monitoring for efficacy and for detection of dangerous sequelae, such as bleeding, thrombosis, or heparin induced thrombocytopenia. PMID- 18396735 TI - Capturing medical data in the EMR. PMID- 18396736 TI - CTS standardization: from donor to recipient. PMID- 18396738 TI - Addressing management issues. PMID- 18396737 TI - The man who invented MLO. PMID- 18396739 TI - The legally tricky e-information age. PMID- 18396740 TI - FDA and clinical labs: beginning a dialogue. PMID- 18396741 TI - History of the Belgian Week of Gastroenterology on the occasion of its XXth manifestation in 2008. PMID- 18396742 TI - A pilot observational survey of hepatitis C in Belgium. AB - AIM OF THE STUDY: There is a lack of epidemiological data on hepatitis C (HCV) infected patients in Belgium. Therefore our purpose was to address this important question and to evaluate the feasibility of a national HCV observatory. PATIENTS AND METHODS: From November 2003 to November 2004, every new patient prospectively seen for HCV antibody positivity in 9 Belgian hospital centres was recorded and a standardised 10-items questionnaire was completed during the consultation, including a Quality of Live (QOL) visual analogue scale. RESULTS: Three hundred and eighteen consecutive patients were recruited. Fifty five percent were male with a median age of 45 y (11-87 y). The main risk factors for infection were IV drug use (27%), blood transfusion (23%), and invasive medical procedure (11%). On the QOL scale, ranging from 0 and 100, mean value was 61 +/- 31. Transaminases were abnormal in 66% with a median elevation 2 times above normal value. HCV RNA was positive in 87% with a viral load above 800 000 IU/ml in 42%. Genotype 1 was predominant (59%), followed by genotypes 3 (19%) and 4 (14%). A liver biopsy was performed in 190 patients, with minimal fibrosis (METAVIR F0-F1) in 43%, moderate fibrosis (F2) in 35% and advanced stages (F3-F4) in 22%. Antiviral treatment was not considered in 53% because of normal ALT (30%), old age (7%), minimal histological stage (6%) or patient refusal (4%). CONCLUSIONS: This study highlights the feasibility of a national HCV survey using a simple questionnaire. This pilot study could be generalised throughout Belgium, and, if repeated, could allow a regular assessment of the changes in epidemiology and management of HCV infection in our country. PMID- 18396743 TI - Incidence and characteristics of type 2 hepatorenal syndrome in patients with cirrhosis and refractory ascites. AB - BACKGROUND AND AIM: Type 2 hepatorenal syndrome (HRS) is a well described progressive impairment of renal function in patients with cirrhosis but its natural history, especially in patients with refractory ascites, is not well known. The aim of this study was to assess the incidence, predictive factors and outcome of type 2 HRS in patients with cirrhosis and refractory ascites. PATIENTS AND METHODS: Thirty patients with refractory ascites were followed-up for 17.5 +/ 26.3 months. The clinical characteristics, biological findings and outcome were analysed. The occurrence of renal dysfunction, and type 2 HRS in particular, was systematically analysed. RESULTS: Twenty-five patients (83.3%) developed renal dysfunction. Type 2 HRS was diagnosed in 16 patients (53.3%). Patients with type 2 HRS were older than patients without (64.8 +/- 9.1 yr vs 52.8 +/- 9.0 yr ; p < 0.001). All the others studied variables were similar between type 2 HRS and non type 2 HRS patients. There was no significant difference in the overall probability of survival between these 2 groups. CONCLUSIONS: In patients with cirrhosis and refractory ascites, the development of type 2 HRS is frequent but does not occur in more severe liver disease and does not affect prognosis. PMID- 18396744 TI - Comparison of standard polyethylene glycol and two doses of oral sodium phosphate solution in precolonoscopy bowel preparation: a randomized controlled trial. AB - BACKGROUND AND STUDY AIMS: This study was undertaken to compare the efficacy, side effects and patient acceptance of standard 4-liters polyethylene glycol (PEG) and 2 doses of sodium phosphate (NaP) solution for precolonoscopy colon cleansing. PATIENTS AND METHODS: A total of 182 patients were randomized to receive either standard 4-L PEG (88 patients) or 80 mL of NaP (94 patients) in a split regimen of two 40 mL doses separated by 24 h, prior to colonoscopic evaluation. The primary endpoint was the segmental assessment of colonic wall visualization. Secondary outcomes included percent of assumed preparation, and the patient tolerance and acceptability. RESULTS: A significantly higher completion rate was found in the NaP group compared to the PEG group (84.3% vs 62.9%; difference, 21.40%; 95% confidence interval [CI], 8.29% to 34.51%; p = 0.001). PEG solution caused more nausea than NaP solution (p = 0.024). Patient acceptance for bowel preparation with NaP was greater (p = 0.019). Adequate colon wall visualization was achieved in similar proportion of patients in both groups with exception of the descending colon, where NaP regimen was superior (72.0% vs 52.9%; difference, 19.10%; 95% CI, 5.20% to 33.00% ; p = 0.012). CONCLUSIONS: Two doses of NaP solution, taken 24 h and 12 h before colonoscopy, tend to guarantee superior results in colonic cleansing with respect to standard 4-liters PEG solution. Taking the second dose of NaP 24 h after the first dose reduces side effects and allows achieving a more satisfactory compliance of the patient. PMID- 18396745 TI - Infliximab in experimental alkali burns of the oesophagus in the rat. AB - BACKGROUND: We aimed to investigate the efficacy of infliximab, a chimeric TNF alpha antibody, in the prevention of fibrosis in an experimental alkaline burn of the oesophagus in the rat. METHODS: Thirty-two Wistar albino rats divided into four experimental groups. Caustic oesophageal burn was induced by applying 37.5% NaOH to the distal oesophagus. Infliximab was given at a dose of 5 mg/kg via the intraperitoneal route. Group A (sham) animals were uninjured, group B had untreated oesophageal burns, group C had oesophageal burns treated with a single dose of infliximab on the first day, and Group D had oesophageal burns treated with infliximab on the first and 14th days. Efficacy of the treatment was assessed on the 28th-day by measuring stenosis index of the oesophagus and histopathological damage score, and biochemically by determining tissue hydroxyproline content. RESULTS: There was no significant difference between the Group B and the infliximab treated Groups C and D in means of tissue hydroxyproline content and histopathological damage scores. Stenosis index was not significantly different between the Group B, Group C, and Group D. CONCLUSION: Anti-TNF-alpha treatment with infliximab does not ameliorate the degree of fibrosis in alkali burns of the oesophagus in the rat. Further evaluation of inflammatory and immunological events leading to stricture in alkaline oesophageal burns may provide new perspectives for the treatment of alkaline oesophageal burns. PMID- 18396746 TI - Necrotizing pancreatitis due to poisoning with organophosphate pesticides. AB - Several complications have been reported in relation to organophosphate poisoning. Pancreatitis due to cholinergic hypersecretion related to this type of poisoning, is however rare and has usually a subclinical course. Necrotizing pancreatitis has only been reported in 3 patients. We present a case of a young man who deliberately ingested the organophosphate dichlorvos and developed a necrotizing pancreatitis. A distal spleen and vessel preserving pancreatectomy was performed already 36 hours after ingestion. We believe that due to this very early surgery, this patient could be discharged as early as 12 days after surgery. PMID- 18396747 TI - Lamivudine treatment for acute severe hepatitis B: report of a case and review of the literature. AB - The oral nucleoside analogue lamivudine has been effectively used in the treatment of chronic hepatitis B. However, there is limited data concerning the efficacy and safety of lamivudine in patients with severe acute or fulminant hepatitis B. We report the use of lamivudine in a young woman with acute HBV infection and fulminant hepatic failure. Following lamivudine treatment, we noticed a prompt clinical, biochemical, serological and virological response as it was seen in the vast majority of, previously reported, cases. Lamivudine treatment was continued until HBsAg was cleared. Our case, as well as previously reported ones, suggests that lamivudine may have a beneficial effect in selected patients with acute severe or fulminant HBV infection. PMID- 18396748 TI - Epstein-Barr virus related lymphoma in inflammatory bowel disease. AB - Epstein-Barr virus (EBV) induced lymphoproliferative disease is a well-known, feared complication of EBV primo-infection in children treated with immunomodulators or immunosuppressive drugs, eg after transplantation. As the incidence of inflammatory bowel disease (IBD) in young children is rising, more young EBV naive patients are treated with immunomodulatory agents. It is not yet clear whether these patients carry the same risk as transplanted patients to develop lymphoproliferative disease and if so, whether their evolution is comparable. We present the history of a young patient with Crohn's disease who developed an EBV related lymphoma shortly after the primo-infection while being treated with azathioprine. This case argues for a rigorous follow up of young IBD patients treated with immune suppressive drugs, also regarding EBV status. PMID- 18396749 TI - Acute hepatitis with prolonged cholestasis and disappearance of interlobular bile ducts following tibolone and Hypericum perforatum (St. John's wort). Case of drug interaction? AB - The case of a patient under tibolone therapy for two years who developed a mixed type liver injury with prolonged cholestasis and features of the vanishing bile duct syndrome following a ten weeks treatment with St. John wort (Hypericum Perforatum) infusions is reported. In the absence of evidence of a potential role for concomitant medication i.e. hydroxychloroquine sulfate to play a role in the clinical, biochemical and morphological picture, an interaction between the herbal preparation and tibolone was suspected as the likely cause of liver damage. PMID- 18396750 TI - Portal vein thrombosis as the first sign of a primary myeloproliferative disorder: diagnostic interest of the V617F JAK-2 mutation. A report of 2 cases. AB - Portal vein thrombosis is, in some cases, related to a myeloproliferative disorder but the diagnosis of the latent forms may be difficult in case of normal blood counts. We report two cases of patient with portal vein thrombosis of unknown origin in whom the presence of the V617F mutation of the Janus Kinase 2 gene lead to the diagnosis of primary myeloproliferative disorder, confirmed on bone marrow examination. The search of the V617F mutation of the Janus Kinase 2 gene has to be performed in all cases of portal vein thrombosis of unknown origin. PMID- 18396751 TI - The risk of surgery in patients with cirrhosis. AB - Several reasons result in the finding that patients with cirrhosis need surgery more often than other patients groups. Patients with cirrhosis frequently have comorbidities resulting in gastrointestinal, lung or cervical cancer, among others. Independent of cirrhosis, surgical resection may be the best alternative for a number of those malignancies. Comorbidities may also result in an increased incidence of vascular complications (such as lower extremity atherosclerosis and coronary stenosis) some of them being potential indications for surgery. Patients with alcoholic cirrhosis are more frequently subjected to trauma and bone fractures. Ascites leads to umbilical hernia which can be strangulated or ruptured. Emergency surgery may be needed in this context. Finally, a significant proportion of patients with cirrhosis develop hepatocellular carcinoma (HCC) during the course of the disease. Surgical resection remains a first line option for HCC. While reliable guidelines have been proposed for surgical resection of HCC and liver transplantation, no precise guidelines are available for other aspects of surgical management during cirrhosis. Specific surgical procedures such as hepatectomy and transplantation are concentrated in highly specialised centres, where detailed evaluation is relatively easy to obtain. In contrast, more general surgical procedures, either abdominal or non abdominal, are performed in various centres, making it more difficult to obtain detailed evaluation and draw recommendations. General surveys are still needed to precisely assess the risk of non-specific surgery in patients with cirrhosis, to identify risk factors and to propose reliable guidelines. PMID- 18396753 TI - [Some aspects worth concern in the management of burn injury]. AB - Although the outcome of burn patients has been improved, many aspects of management of severe burn patients remain controversial. Here we focus on the management of hypermetabolism and the resuscitation of respiratory function. Currently, the fluid resuscitation method shifts from insufficient fluid regimen to excessive fluid loading. The benefit of colloid infusion and restrictive blood transfusion need to be authenticated by further clinical trial, and the best form of fluid resuscitation has yet to be identified. The respiratory management of burn patients had been improved. Early tracheostomy, ventilation with low tidal volume and bronchoalveolar toilet are recommended. Many potential beneficial treatment strategies have been identified by recent research in the metabolic response to burn injury. Although immunomodulation therapy is promising, most of them are not clinical viable,and further clinical research is warranted. PMID- 18396752 TI - The metabolic syndrome and the liver. AB - Together with the worldwide epidemic proportions of obesity the incidence of 'the metabolic syndrome' is rising across countries. The metabolic syndrome is described as a complex condition that is linked to (intra-abdominal) obesity and is characterized by insulin resistance, dyslipidaemia and hypertension. Several definitions for the metabolic syndrome have been suggested, all trying to identify individuals at high risk for both type 2 diabetes and cardiovascular disease. The primary hepatic complication of obesity and insulin resistance is nonalcoholic fatty liver disease (NAFLD). NAFLD is not included as a component of the metabolic syndrome as it is currently defined; however, data suggest an association. Although the data are mainly epidemiological, the pathogenesis of NAFLD and the metabolic syndrome show common components, with the focus on insulin resistance as a key factor. Even so the treatment of patients with the metabolic syndrome and NAFLD shows a certain degree of similarity, and should focus on the management of associated conditions including obesity, glucose and lipid abnormalities. Lifestyle modifications comprising healthy eating habits and regular exercise are the primary interventions recommended to patients with the metabolic syndrome and those with NAFLD. A pharmacological approach like insulin sensitizing agents, lipid lowering drugs, antihypertensive drugs and antiobesity agents can be successful in the treatment of certain risk factors that are currently clustering with both the metabolic syndrome and NAFLD. In some cases bariatric surgery may be necessary. PMID- 18396754 TI - [Relationship between the endothelial barrier and vascular permeability after burns and its mechanism]. AB - Massive burn trauma is characterized by hypovolemic shock induced by the loss of plasma from vessels. The major reasons for this systemic microvascular leakage in burns include an increase in vascular permeability triggered by inflammatory mediators and the increase of vascular hydrostatic pressure caused by vessel dilation. The maintenance of normal vascular permeability depends on the integrity of endothelial barrier function regulated by the interaction of intracellular junctions, cell-matrix adhesion and the cytoskeleton contractile force. This review summarizes some recent discovery in endothelial mechanisms during burn-induced vascular hyperpermeability. PMID- 18396755 TI - [Protective effect of nitric oxide production on myocardium in severely scalded rats]. AB - OBJECTIVE: To observe the influence of aminoguanidine on cardiac troponin (cTnI) and nitric oxide (NO) levels in serum and myocardium in severely scalded rats. METHODS: Seventy-two Wistar rats were subjected to 30% TBSA full-thickness scald and randomly divided into scald group(S) and aminoguanidine group (A, with intraperitoneal injection of 40 mg/kg aminoguanidine before scald). The venous blood and myocardial tissue of the rats were harvested for the determination of the level of cTnI and nitrite in both serum and myocardium before scald and at 1, 3, 6, 12 and 24 post-burn hours(PBH). Six sham scalded rats served as control group. The changes in the cTnI level and myocardial function were determined among control group, A and S groups at 6PBH. RESULTS: The serum level of NO in S group [(59.6 +/- 5.4) micromol/L] was obviously higher than that before scald [(24.6 +/- 0.8) micromol/L, P < 0.01], and it peaked at 6 PBH, then decreased obviously at 24 PBH, which was still markedly higher than that in A group (P < 0.01). The changes in NO level in myocardium were similar to the above tendency. Compared with S group, the level of cTnI was significantly increased in A group at each time-point. Compared with A group at 6 PBH, the inhibition of the cardiac function was relatively reduced in S group at 6 PBH. CONCLUSION: Inhibition of NO synthesis by aminoguanidine aggravates cardiac damage and impairment of cardiac function of scalded rats, indicating that NO exerts protective effect on myocardium at early stage after a scald injury. PMID- 18396756 TI - [Influence of heat shock factor 1 gene transfection on the expression of inflammatory mediators in macrophages induced by burn serum]. AB - OBJECTIVE: To investigate the influence of heat shock factor1 (HSF1) on gene expression of inflammatory mediators in RAW264.7 murine macrophage cells induced by burn serum. METHODS: Sera were separated from blood of normal rats and rats with severe burns, and the recombinant vector pcDNA3. 1/HSF1 was constructed. RAW264.7 macrophages were divided into non-transfection group, vacant vector group (with burn and normal sera stimulation, respectively after vacant vector transfection) and recombinant vector group (with burn and normal sera stimulation, respectively after recombinant vector transfection). Some recombinant vector transfected macrophages without serum stimulation were prepared for the determination of HSF 1 expression with Western blotting. The mRNA expressions of TNF-alpha, HMGB 1 and IL-10 were determined with RT-PCR. RESULTS: The cell line attained after recombinant vector transfection was comparatively stable,with partial activation of HSF 1. Burn sera markedly upregulated TNF-alpha, HMGB1 mRNA expression (0.910 +/- 0.100, 0.860 +/- 0.020, respectively), but downregulated IL-10 expression (0.430 +/- 0.010, respectively) in normal macrophages, while these genes maintained in a very low level in normal macrophages with normal serum stimulation . macrophages with recombinant vector transfection and burn serum stimulation could obviously inhibit the expression of TNF-alpha and HMGB 1, but enhance the IL-10 gene expression (0.130 +/- 0.100, 0.450 +/- 0.020 , 0.450 +/- 0.020, respectively )when compared with that with vacant vector transfection and burn serum stimulation (0.800 +/- 0.050, 0.880 +/- 0.030, 0.420 +/- 0.010, respectively). CONCLUSION: HSF1 can inhibit the expression of some pro-inflammatory mediators in macrophages after a severe burns, indicating that appropriate upregulation of anti-inflammatory mediators might exert protective effects on the organism. PMID- 18396757 TI - [Protective effects of administration of enalapril maleate on rat myocardial damage in early stage of burns]. AB - OBJECTIVE: To investigate the preventive and therapeutic effects of enalapril maleate (Enalaprilat) (E) on myocardial damage in early stage after burns. METHODS: A total of 60 SD rats were subjected to 30% TBSA III degree scald injury, and randomly divided into scald group (with conventional fluid transfusion after scald) and ENA group (with intraperitoneal injection of 1 mg/kg Enalaprilat after scald). Normal control consisted of 6 rats. Plasma levels of cTnI and CK-MB were determined in all the groups at 1, 3, 6, 12, 24 post-scald hours (PSH) by enzyme linked immunosorbent assay. The pathological changes in myocardium were observed at the same time-points. RESULTS: (1) The serum level of cTnI and CK-MB in scald group were significantly higher than that of normal controls at each time-point (P < 0.01). The serum level of cTnI and CK-MB in ENA group were (1.32 +/- 0.12 microg/L to 2.47 +/- 0.22 microg/L) and (438 +/- 68 U/L to 5569 +/- 322 U/L), respectively, which were obviously lower than those in B group (6.42 +/- 0.96 microg/L to 15.10 +/- 3.69 microg/L) and (2556 +/- 74 U/L to 8047 +/- 574 U/L, P < 0.05 or P < 0.01) at different time-points. (2) Compared with normal controls, cloudy swelling, stromal blood vessel dilatation and congestion inflammatory cell infiltration were observed in scald group, but these pathological changes were less marked in ENA group. CONCLUSION: Severe myocardial damage in rat occurred early after burns. Enalaprilat injection can markedly alleviate myocardial damage. PMID- 18396758 TI - [Comparison of the burn wound and diabetic ulcer wound]. AB - OBJECTIVE: To compare the difference between the burn wound and diabetic ulcer wound, and to preliminarily analyze the nonhealing mechanism of diabetic unclear. METHODS: The tissue of foot ulcer of diabete patients and skin wound tissues from burn patients were harvested. The levels of (FGF)2 and VEGF in the wound tissues were determined after tissue cultivation with enzyme-linked immunosorbent assay (ELISA). The changes in micro-vascular density (MVD) were examined by immunohistochemistry. Human umbilical vein endothelial cells were cultured in medium containing different components, and divided into following groups: A (with treatment of 5 mmol/L glucose for 7 days), B (with treatment of 30 mmol/L glucose for 7 days) and C (with treatment of 30 mmol/L Mannitol for 7 days) groups, then the level of VEGF protein was determined by ELISA. RESULTS: The levels of FGF2 and VEGF protein in the burn wound were (59 +/- 3) ng/ml and (56 +/- 7) pg/ml, respectively, which were obviously lower than those in diabetic ulcer wound [(89 +/- 6) ng/ml, (108 +/- 5) pg/ml, P < 0.05]. There was also obvious difference in MVD between two kinds of wound (P < 0.05). The level of VEGF protein in both wounds were similar after the addition of FGF2 to the cell culture in vitro, while there were statistically significant difference 2 and 5 days after removal of FGF. CONCLUSION: The nonhealing mechanism of diabetic ulcer wound may be related to the inhibition of vacuolation and low expression of factors controlling vessel growth. PMID- 18396759 TI - [Ectopic expression of keratin 19 and integrin beta1 during wound healing process after microskin grafting in rats]. AB - OBJECTIVE: To observe the characteristics of keratin 19 and integrin beta1 expressions in the wound after microskin grafting , and to investigate the healing mechanism. METHODS: Full layer skin defects were created in twenty Sprague-Dawley rats and they were divided into two groups, i.e., A group (with grafting of autologous microskin accounting 10% in weight of epidermis loss from skin defect), B group (with grafting of autologous microskin and allogeneic microskin, accounting 10% and 40% weight of epidermis loss respectively in skin defect). The wound healing rate and contraction rate were observed at 2,3,4 post grafting week (PGW), and the expression and distribution of keratin 19 and integrin beta1 were observed at 2 and 4 PGW. RESULTS: The wound healing rate in the B group on 2 and 3 PGW was obviously higher than that in A group [(85 +/- 5)% vs. (53 +/- 10)%, (84 +/- 8)% vs. (65 +/- 9)%, P < 0.01]. No obvious difference in wound contraction rate between the two groups was observed on the 2, 3 and 4 PGW (P > 0.05). Cells with expression of keratin 19 and integrin beta1 were observed in the suprabasal layers of the epidermis in healing wound, but not in the basal membrane. Integrin beta1 positive expression cells were not observed in the suprabasal layers until 4 PGW. CONCLUSION: Mixed grafting with autogenous and allogenous microskin can improve wound healing. Ectopic expression of keratin 19 and integrin beta1 exists during wound healing process after microskin grafting. PMID- 18396760 TI - [The influence of terbutaline on VEGF gene expression in rat astrocytes after norepinephrine and burn serum induction]. AB - OBJECTIVE: To investigate the influence of adrenoreceptor beta-agonists terbutaline on gene expression of vascular endothelial growth factor (VEGF) in rat astrocyte after induction by norepinephrine (NE) and burn serum. METHODS: The sera of normal and burn rats were separated for use. Primary astrocytes of brain tissue were isolated from neonatal 1-3 d rats and cultured and divided into following groups: (1) CONTROL GROUP: with 10% normal rat serum in the culture medium. (2) NEl, NE2, NE3 groups: with 10% burn rat serum and 10, 20, 50 micromol/L NE in the culture medium, respectively. (3) TBN1, TBN2, TBN3 groups: with 10% burn rat serum and 10, 20, 50 micromol/L NE and 10, 20, 50 micromol/L terbutaline in the culture medium, respectively. The mRNA and protein expression of VEGF in each group were determined with real-time PCR and Western blotting, respectively. RESULTS: There was a low protein expression of VEGF in control group, but it increased slightly in NE1 group, and then increase gradually in NE2, NE3 groups, and it was obviously increased in TBN1, TBN2, TBN3 groups. The mRNA expression of VEGF in NE1, NE2, NE3 groups were [(13.26 +/- 0.03), (10.37 +/ 0.04), (14.87 +/- 0.55) copies/g], respectively, which were significantly higher than that of control [(5.72 +/- 0.12) copies/g, P < 0.01]. In addition, the expression of VEGF mRNA in TBN1, TBN2, TBN3 groups was higher than that in control group, and expression of VEGF mRNA [(13.39 +/- 0.19), (15.77 +/- 0.11), (16.00 +/- 0.07) copies/g] was gradually increased, which showed obvious difference between TBN2 and NE2, and also between TBN3 and NE3 groups (P < 0.01). CONCLUSION: Terbutaline can increase gene expression of VEGF in rat astrocytes after induction by NE and burn serum. PMID- 18396761 TI - [Survey on the distribution of burn pathogens and their antibiotic resistance in burn unit]. AB - OBJECTIVE: To investigate the distribution of burn pathogens and their antibiotic resistance in a burn unit, so as to provide reference for clinical practice. METHODS: Three hundred and forty-eight burn patients hospitalized in our department were enrolled in this study. The pathogens isolated from the wounds, blood, venous catheter, sputum, urine, purulent discharge of wounds in these patients, and their antibiotic resistance were surveyed by retrospective analysis from Jan, 2001 to Dec, 2006. RESULTS: Total-ly 464 strains were isolated, among which Gram negative (G-) bacilli accounted for 52.6%, Gram positive microorganisms (G+) accounted for 40.5%, and fungi accounted for 6.9%. The main pathogens were Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter species and Escherichia coli, among which Staphylococcus aureus (MRSA) was predominant (93.5%). MRSA was 100% resistant to levofloxacin, penicillium, oxacillin, and it was also resistant to other antibiotics except Vancomycin. The resistance rate of Pseudomonas aeruginosa to Cefoperazone/Sulbactam, Imipenem and cefepime were 15.8%, 36.8%, 33.3%, respectively. CONCLUSION: Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter species and Escherichia coli were predominant in the burn unit,among them Staphylococcus aureus and Acinetobacter were more resistant to antibiotics. PMID- 18396762 TI - [Preliminary study of the ultrasonic measurement of thickness of skin in children]. AB - OBJECTIVE: To collect the data of measuring skin thickness of children of both genders of different ages and parts of body with non-invasive high-frequency ultrasound method. METHODS: Two hundred and twenty-one children from 1 to 18 years of age,without systemic disease or injury in skin, were enrolled in the study and divided into 4 groups: i.e., infant group (112 years of age), pre school age group (3-6 years of age), school age group (7-12 years for boys and 7 11 years for girls), adolescent age group (13-18 years for boys and 12-18 years for girls), and each group was subdivided into 2 groups according to the gender. The skin thicknesses of children in cheek, chest, abdomen, forearms, fundament and thigh was respectively measured by 13 MHz high-frequency ultrasound. RESULTS: The region with thinnest skin in children was the cheek, and the thickest was the back and buttock. (1) There were no significant differences in thickness of skin in the same region between genders and also among different age groups (P > 0.05). (2) There were also no obvious differences of thickness of the dermis and the whole skin in the same region between male and female, or among infants, pre school age and school age groups (P > 0.05). In adolescent group, the average thickness of dermis in male was (1.16 +/- 0.04 ) - (1.98 +/- 0.47) mm, the average whole thickness of skin in male was (1.27 +/- 0.12) - (2.20 +/- 0.45) mm, while those of female were (1.00 +/- 0.18) - (1.60 +/- 0.30) mm and (1.10 +/- 0.17) - (1.83 +/- 0.29) mm (P < 0.05). CONCLUSION: It is reliable to measure the skin thickness by 13MHz ultrasound as a non-invasive method. The main factor which determined the thickness of the skin is dermal thickness, especially in males. The significant differences of skin thickness among cheek, back and buttock provide the basis for us to choose the appropriate thickness of skin grafts harvested from different body parts. PMID- 18396763 TI - [Application of reverse island flaps pedicled with cutaneous nerve nutrient vessels to repair defect on distal parts of extremities]. AB - OBJECTIVE: To analyze the feasibility of reverse island flaps pedicled with cutaneous nerve nutrient vessels in repairing the defect on distal parts of extremities. METHODS: Thirty patients with tissue defect in distal extremities, complicated by exposed vessels, nerve, tendons, and/or bones ,were repaired with island flaps pedicled with neurocutaneous nutrient vessel. Among them, five cases were grafted with flaps with external forearm cutaneous nerve vessels, eleven cases were grafted with flap containing sural neural nutrient vessels in its pedicle,and 14 cases with saphenous nerve nutrient vessels in the pedicles. RESULTS: The flaps of 28 cases survived with perfect appearance and function. In one case partial necrosis occurred because of compression on the pedicle, but the patient healed after dressing, and another case with necrosis of the edge of the flap due to infection, but also healed after skin grafting. CONCLUSION: The reverse island flaps pedicled with cutaneous nerve nutrient vessels which are constant in anatomy, with reliable blood supply, can be recommended because its simple operative technique, non-injurious to main blood vessels and muscles, the repair of distal defects of the extremities. PMID- 18396764 TI - [Analysis of drug use of 290 burn patients]. AB - OBJECTIVE: To analyze the drug use in our burn ward, and to investigate the methods in controlling the proportion of drug expenses, as well as the rational use of antibiotics. METHODS: A randomized sample of 290 burn-patients with hospital stay more than 5 days in 2005 were enrolled in the study, and information about their drug use, (especially that of antibiotics), the drug expenses and its proportion in the total medical expenses, the total therapeutic expenses, and the healing rate of the patients were statically analyzed. RESULTS: Sulfamido was dominant among the topically applied antibiotics, while cephalosporins was dominant among the systemic applied antibiotics. The drug expenses accounted for (11 +/- 5)% in the therapy expenses, while the antibiotics expenses accounted for (5.8 +/- 1.7)% in the therapy expenses, but it accounted for (51 +/- 17)% in the drug expenses. The average expense of every patient was 22026.09 RMB, and the healing rate of the patients was 96.90%. CONCLUSION: The proportion of drug expense (especially that of antibiotics), as well as total medical expenses of burn patients can be lowered through a combined therapy. PMID- 18396765 TI - [Categorization of burn severity]. AB - OBJECTIVE: To seek a new method for the categorization of burn severity. METHODS: Burn patients hospitalized in our center from December of 1958 to December of 2004 were enrolled in the study, and they were divided into different age groups according to same mortality, then the patients in each group were subdivided into 4 groups according to the burn severity: i.e., mild burns, moderate burns, severe burns, serious severe burns. The total burn area, the number of cases, the mortality, and the area of DI degree burns were statistically analyzed in each subgroup, and the scope in total burn area and area of III degree burns were taken as standards to define the degree of burns. The logistic regression equation was established with probability of death as the variable, and age, total burn area, burn area of different degrees as concomitant variables to form a logistic regression formula. It was used to predict the probability of death of patients hospitalized in 2005, 50 as to check whether the corresponding indices of these patients were consistant with above standard of categorization into degrees, and to judge hum severity of the patients who had concomitant inhalation injury, severe associated injury, or those with serious disease before burns. RESULTS: The patients were divided into three groups: less than 2 years of age (including 2 years of age), 2 to 55 years of age(including 55 years of age), and older than 55 years of age groups. The classification standard of burn area was shown in table 2 of the article. The probability of death and corresponding indices predicted hy the logistic regression equation were highly coincident with our standard. Patients with moderate inhalation injury could be regarded as patients with severe or most severe burns, while severity of those with mild inhalation injury could be determined by burn area alone. CONCLUSION: The logistic regression equation is a good method to predict the severity of burn patients, with reasonable age specificity grouping, and accurate and practical scoring of division for corresponding burn severity. PMID- 18396766 TI - [Statistical analysis of articles, citations and authors in Chinese Journal of Burns from 2003 to 2005]. AB - OBJECTIVE: To study the characteristics of articles, citations and authors in Chinese Journal of Burns (Chin J Burns). METHODS: The date concerning articles, citations and authors of Chin J Burns from 2003 to 2005 were collected and analyzed in terms of bibliographic parameters. RESULTS: There were 741 papers published in Chin J Burns during these 3 years, and papers with funded projects accounted for 19.7%. The average number of articles per issue was 41,with paper density of 0.59, and publication date were 15 months in average after submission. Among these papers, 87.9% of which were shared with co-authors, with a corporation rate of 3.94. The papers with citation accounted for 78.7%, and 90.6% of all the citations came from journals, especially English journals. The Price's indicator was 65.6%. CONCLUSION: Chinese Journal of Burns, an academic journal of high quality, with strong influence, pushes the discipline of Burns medicine in China forward. PMID- 18396767 TI - [Histologic characteristics of in vitro constructed skin basement membrane]. AB - OBJECTIVE: To observe the histological characteristics of constructed basement membrane in tissue-engineered skin. METHODS: Forskins from circumcision in normal children were obtained with informed consent of the parents, and then the epidermal keratinocytes (KC) and dermal fibroblasts (Fb) were isolated with trypsin and collagenase D digestion in sequence. Tissue engineered skin with composite chitosan was maintained in a submerged state for 3 days, and then at the air-liquid interface. The tissue-engineered skins were fixed in neutral formalin and then embedded in paraffin after culture for 7, 10 and 15 days, respectively for immunohistological examination of the basement membrane component,including the condition of collagen type IV (COL-IV), collagen type VII (COL-VII), and laminin (LN). RESULTS: HE staining showed that the keratinocytes formed a fine stratified squamous epithelium with the presence of basal, spinous, granular and corneous cell layers, and there was various amount of cells in flat and fusiform shape in each layer. It was found that a regular red staining strip situated at the dermal epidermal junction. Positive staining of collagen IV, collagen VII as well as LN was observed by immunohistological examination. CONCLUSION: The results suggest that the composite chitosan tissue engineered skin has a good prospect for clinical use because it presents a perfect reconstruction of basement membrane. PMID- 18396769 TI - Worse than the BCS. PMID- 18396768 TI - Tag, you're it. PMID- 18396770 TI - Spread too thin. PMID- 18396771 TI - Beyond 15. PMID- 18396772 TI - Top heart surgeons Cooley and DeBakey put their decades-old feud to rest. PMID- 18396773 TI - Simulation training in a pediatric residency program. PMID- 18396774 TI - TGF-beta antisense oligonucleotides modulate expression of matrix metalloproteinases in isolated fibroblasts from radiated skin. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) has been identified as an important component of wound healing. Recent developments in molecular therapy offer exciting prospects for the modulation of wound healing, specifically those targeting TGF-beta. The purpose of this study was to analyze the effect of TGF beta targeting on the expression of matrix metalloproteinases (MMPs) in fibroblasts isolated from radiation-induced chronic dermal wounds. MATERIALS AND METHODS: The expression of MMPs in tissue samples from radiation-induced chronic dermal wounds was investigated by immunohistochemistry and microarray technique. The effect of TGF-beta targeting using antisense oligonucleotides on the expression of MMPs in isolated fibroblasts was analysed by ELISA and multiplex RT PCR. RESULTS: Immunohistochemical investigation and microarray analysis demonstrated an increased expression of MMP protein and mRNA in tissue samples from radiation-induced chronic dermal wounds compared to normal human skin. Antisense TGF-beta oligonucleotide treatment significantly down-regulated MMP secretion in vitro. CONCLUSION: TGF-beta antisense oligonucleotide technology may be a potential therapeutic option for the inhibition of proteolytic tissue destruction in radiation-induced chronic wounds. PMID- 18396775 TI - The frequency of gastric pyloric cells with glassy cytoplasm in baboons. A comparison with human subjects. AB - BACKGROUND: Pyloric cells with "glassy" cytoplasm were detected 20 years ago in the human gastric mucosa, and subsequently in the gastric mucosa of two baboons and of transgenic mice. These pyloric cells, called glassy cells (GCs) show a homogeneous, eosinophilic material that pushes the nucleous towards the free border of the cell. MATERIALS AND METHODS: The frequency of baboons having pyloric glands with GCs was assessed in the stomachs of 92 consecutive animals, dying of non-gastrointestinal disorders. RESULTS: High power examination of the apparently normal gastric mucosa revealed that 10.9% (n=10) of the 92 baboons had GCs. Periodic acid Schiff (PAS) and PAS-diastase stains suggested that the glassy material might be a glycoprotein. CONCLUSION: The relatively high frequency of GCs in the gastric mucosa of baboons suggests that this animal might be a useful model for investigating the effect of the environment in the development of GCs, as well as ascertaining the true nature of the retained "glassy" material. PMID- 18396776 TI - Suppressive activity of epinastine hydrochloride on eosinophil activation in vitro. AB - The influence of a histamine H1 receptor antagonist, epinastine hydrochloride (EP), on eosinophil functions was examined in vitro and in vivo. The first set of experiments was undertaken to examine whether EP could suppress eosinophilia and IgE hyperproduction induced by Mesocestoides cortii infection in BALB/c mice. The number of peripheral blood eosinophils and levels of IgE were examined 21 days after infection. Oral administration of EP at a daily dose of 0.3 mg/kg, which is the recommended human therapeutic dose, for 21 days was not able to suppress either peripheral blood eosinophilia or IgE hyperproduction, which was observed in mice infected with M. cortii. The second part of the experiment was designed to examine the influence of EP on eosinophil activation induced by stem cell factor (SCF) stimulation in vitro. Eosinophils were obtained from M. cortii infected mice and stimulated with SCF in the presence of different concentrations of EP for 24 h. The addition of EP into cell cultures suppressed eosinophil activation induced by SCF stimulation as assessed by measuring the contents of acronym for Regulated upon Activation, Normal T cell Expressed and presumably Secreted (RANTES), macrophage inflammatory protein-1beta (MIP-1beta) and leukotriene C4 (LTC4) levels in culture supernatants. The minimum concentration of EP which caused significant suppression of factor productions was 25 ng/ml, which is similar to the concentration in plasma after oral administration of the therapeutic dose in humans. These results may suggest that EP exerts inhibitory effects on eosinophil activation and results in favorable modification of the clinical status of allergic patients. PMID- 18396777 TI - Growth inhibition and induction of apoptosis by 2-methoxyestradiol in rat osteosarcoma and malignant fibrous histiocytoma cell lines. AB - 2-Methoxyestradiol (2-ME) has been found to possess antitumor activity in vivo and in vitro. It has been suggested that 2-ME induces apoptosis resulting in G2/M arrest of tumor cells. In this study, the effect of 2-ME was evaluated in rat osteosarcoma and malignant fibrous histiocytoma (MFH) cell lines. 2-ME was used at final concentrations of 100 nM to 2 microM. The effect of 2-ME on cell growth was measured by the MTS assay. Induction of apoptosis and activation of caspase-3 were investigated along with apoptosis-related gene expression. The data showed that 2-ME significantly inhibited cell growth, inducing apoptosis. The activity of caspase-3 was increased at 20 h and 40 h in both cell lines. 2-ME induced p16 expression, which was possibly involved in the apoptotic process. These results suggested that the 2-ME-induced apoptosis of rat osteosarcoma and rat MFH cells was accompanied by caspase-3 activation through p16 induction. PMID- 18396778 TI - Characterization of a rabbit antihuman mechano growth factor (MGF) polyclonal antibody against the last 24 amino acids of the E domain. AB - The human insulin-like growth factor-1 (IGF-1) gene gives rise to multiple, heterogeneous mRNA transcripts by alternative splicing, thus producing different IGF-1 isoforms. The mechano growth factor (MGF) is an IGF-1 isoform that was found to be markedly up-regulated in exercised or damaged muscle. The specific E domain of the MGF splice variant may act as an independent growth factor. The aim of the present study was to characterize a rabbit antihuman MGF polyclonal antibody. New-Zealand rabbits were immunized by injections of a purified synthetic peptide corresponding to the last 24 amino acids of the human C terminal of the MGF E domain. Western blotting and immunohistochemical techniques were used to characterize the specificity of the polyclonal anti-MGF antiserum. The anti-MGF antiserum was found to recognize the MGF E-peptide and not the common part of the IGF-1 isoforms, i.e. the mature IGF-1 peptide. Furthermore, it specifically bound to the MGF protein in human skeletal and in rat cardiac muscle, apparently due to the considerable homology between the human and rat MGF E-peptide sequences. Immunostaining analysis showed that this polyclonal anti-MGF antibody was able to detect MGF in human muscle and in rat cardiomyocytes and vessels' smooth muscle cells. We conclude that this rabbit polyclonal anti human/rat MGF antibody could become a valuable tool in the study of IGF-1 isoforms in human and rat tissues. PMID- 18396779 TI - Changes of glucocorticoid receptor expression in the nasal polyps of patients with chronic sinusitis following treatment with glucocorticoid. AB - Glucocorticoids (GCs) bind to the cellular glucocorticoid receptors (GRs) to exert anti-inflammatory and immunosuppressive actions. We investigated the changes in the expressions of the two GR isoforms, GR-alpha and GR-beta, in nasal polyps treated with GC. Immunofluorescent staining revealed prominent expression of GR-alpha in the inflammatory cell infiltrate in the polyps obtained from patients with chronic sinusitis and bronchial asthma. Furthermore, while the expression of GR-alpha was significantly reduced following GC treatment, that of GR-beta remained unchanged. The results of real-time PCR also revealed that the prominent expression of GR-alpha mRNA in the polyps decreased following GC treatment, while the expression of GR-beta mRNA remained unchanged. The observations indicate that GR-alpha may play the major role in the inflammation associated with nasal polyps and the ratio of the expression level of GR-beta to that of GR-alpha may serve as a useful index of the clinical efficacy of GC treatment. PMID- 18396780 TI - Cutaneous T-cell lymphoma in an African hedgehog (Atelerix albiventris). AB - A three-year-old male African hedgehog was presented for a non healing crusty proliferation on the left pinna. The lesion failed to respond to topical therapy and systemic antibiotic therapy. Whole body radiography and abdominal ultrasonograpy were within normal limits. The lesion was surgically removed. The patient recovered well from the procedure and remained in remission for nine months when he came back as an emergency case and died of an unrelated disease. The histopathology report enabled a diagnosis of completely excised cutaneous T cell lymphoma. This report represents the first successful treatment of a cutanous T-cell lymphoma in this species and might help to plan future therapies. PMID- 18396781 TI - Adjuvant electrochemotherapy for incompletely excised anal sac carcinoma in a dog. AB - Canine anal sac gland carcinoma (ASGC) is a frequently described neoplasm that is highly aggressive and can frequently lead to metastatic spread. In this paper, we describe the successful treatment of an incompletely excised ASGC by using cisplatin selectively driven within the tumor cells by trains of biphasic pulses. The dog received two courses of electrochemotherapy 14 days apart. Neither systemic nor local toxicities were detected during the whole course of therapy. The dog is still in complete remission after 18 months. Electrochemotherapy is a safe and efficacious adjuvant therapy for ASGC and warrants further investigation in order to standardize its protocols. PMID- 18396782 TI - Successful rescue of an apocrine gland carcinoma metastatic to the cervical lymph nodes by mitoxantrone coupled with trains of permeabilizing electrical pulses (electrochemotherapy). AB - Canine apocrine gland carcinoma is a locally aggressive neoplasm that can occasionally lead to metastatic spread, thus mimicking the behavior of their human counterpart. In this paper we describe the successful treatment of a cervical metastatic spread of this neoplasia by using mitoxantrone selectively driven within the tumor cells by trains of biphasic pulses. The dog experienced tumor reduction from the first cycle of electrochemotherapy (ECT) and complete remission by the time of its fourth session. Neither systemic or local toxicities were detected during the whole course of therapy. The dog is in complete remission after six months from his last treatment. Electrochemotherapy is a safe and efficacious therapy for metastatic carcinoma and warrants further investigation. PMID- 18396783 TI - Osteopontin expression in human decidua is associated with decidual natural killer cells recruitment and regulated by progesterone. AB - The involvement of the chemokine osteopontin has been proposed for maintaining successful pregnancy in mice and ruminants; however, little information of its function in human pregnancy is available. Osteopontin expression was assessed in decidua by RT-PCR and immunohistochemical staining in early pregnant women and RPL (recurrent pregnancy loss) patients. Osteopontin was expressed both in human decidual stromal cells and decidual natural killer (dNK) cells, and higher expression was detected in the later gestational phase compared to the early gestational phase. The osteopontin expression increased with pregnancy progression and higher osteopontin expression was correlated with a larger number of dNK cells. Compared with normal pregnancy, osteopontin expression and dNK cells accumulation were reduced significantly in RPL patients. Osteopontin expression was regulated by progesterone via an in vitro culture model. Our results indicated that osteopontin may play an important role in dNK recruitment and is an essential factor for successful pregnancy. PMID- 18396784 TI - Curcumin inhibits WEHI-3 leukemia cells in BALB/c mice in vivo. AB - Curcumin (1, 7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5- dione), a natural polyphenol product of the plant Curcuma longa, exhibited potent inhibitory activities against proliferation, induced cell cycle arrest and exhibited the induction of apoptosis in several tumor cell lines. In our previous studies, we have shown that curcumin induced cell cycle arrest and apoptosis on human leukemia HL-60 and mouse leukemia WEHI-3 cells; there are no reports regarding whether or not it affects leukemia cells in vivo. In the present study, we investigated the effects of curcumin on WEHI-3 in BALB/c mice and the results indicated that curcumin reduces the percentage of Mac-3 marker, which is the precursor of macrophage. Curcumin induced significant effects on the population of B cells from murine leukemia in vivo. We also investigated the weights of spleen and liver from murine leukemia and the results showed that curcumin reduced the weight of the liver and spleen. From the pathological examinations, the effects of curcumin on the liver and spleen from mice after being injected with WEHI-3 cells were apparent. Both organs were enlarged. In conclusion, curcumin affect WEHI-3 cells in vivo. PMID- 18396785 TI - Inhibitors of Ca2+ and K+ transport enhance intracellular killing of M. tuberculosis by non-killing macrophages. AB - BACKGROUND: Human monocyte-derived macrophages that have little killing activity of their own kill intracellular Staphylococcus aureus when cultured in the presence of inhibitors of calcium and potassium efflux pumps. The aim of this study was to evaluate the effect of inhibitors such as ouabain, reserpine and verapamil in the killing activity of macrophages infected with Mycobacterium tuberculosis. MATERIALS AND METHODS: Macrophages obtained from peripheral blood were infected with M. tuberculosis ATCC27294 H37Rv and treated with reserpine, ouabain and verapamil. RESULTS: After three days post-infection, macrophages treated with the inhibitors demonstrated an enhancement of the killing activity destroying the internalized bacteria. CONCLUSION: Whereas drugs that target the bacterium are predicted to lose effectiveness due to mutation of the bacterial target, drugs that enhance killing by macrophages that normally do not kill mycobacteria may yield a more effective form of infections therapy caused by multidrug resistant M. tuberculosis. PMID- 18396786 TI - VCAM-1 increases levels of HGF in eosinophilic chronic rhinosinusitis cell culture. AB - BACKGROUND: Eosinophilic chronic rhinosinusitis (ECRS) is one of the most common diseases worldwide. To date the underlying cause remains unclear and no drug has been accredited for first-line therapy. VCAM-1 has been reported to play a pivotal role in establishing ECRS. Other authors have reported that inflammatory cytokines may mediate changes in the underlying epithelium in the sinuses through hepatocyte growth factor HGF. In our study, the effect of VCAM-1 on HGF levels was investigated. MATERIALS AND METHODS: ECRS cell cultures were incubated with VCAM-1 and HGF levels were determined after 16, 24, 48 and 72 hours. RT-PCR was enrolled to depict the HGF-RNA levels. RESULTS: Sixteen hours of incubation showed 28.5 pg/ml HGF, whereas in the control 16.3 pg/ml was detectable. After 24 and 36 hours, 37 pg/ml and 43.5 pg/ml HGF were measured in the incubated cell cultures, respectively; 72 hours of incubation with VCAM-1 resulted in 50.6 pg/ml HGF, whereas 23.5 pg/ml was determined in the controls. The RT-PCR for HGF also showed increased concentration in the incubated cells after 72 hours. CONCLUSION: VCAM-1 induced an increase in levels of HGF in the ECRS cell cultures. The rising transcriptional activity was demonstrated by means of RT-PCR. The levels of HGF were within physiological ranges, suggesting that a misbalance between HGF and VCAM-1 resulted in the establishment of ECRS. Further experiments are necessary to reveal the role of HGF in the development of ECRS. This is the first report about the effect of VCAM-1 on growth factors in ECRS cell culture. PMID- 18396787 TI - Expression in childhood primary brain tumors of NY-ESO-1, a cancer/testis antigen: an immunohistochemical study. AB - BACKGROUND: NY-ESO-1 is a human gene that codes for antigens that are expressed in malignancies of various histological types, but not in normal tissues, except the testes. The expression of NY-ESO-1 in intracranial brain tumors including astrocytomas (ASTRs) and medulloblastomas (MEDs)/primitive neuroectodermal tumors (PNETs) was examined since the expression of NY-ESO-1 has only previously been explored in depth in neuroblastomas. MATERIALS AND METHODS: During our immunohistochemical study, a sensitive, four-step, alkaline phosphatase conjugated antigen detection technique was employed. The expression of NY-ESO-1 was thereby examined in 6 cases of MED/PNET and 14 cases of ASTR. RESULTS: All 6 MED/PNET cases demonstrated high levels of immunoreactivity (overexpression) with the highest immunostaining intensity grades A and B. In the astrocytic tumors of various subtypes examined, the level of NY-ESO-1 expression was not as strong as that in MEDs/PNETs. However, there was a significant increase in expression level when comparing low-grade pilocytic ASTRs to high-grade anaplastic ASTRs and glioblastomas. CONCLUSION: As evidenced by our results, NY-ESO-1 overexpression increases as the malignancy grade of the astrocytic tumors increases. These data suggest that antigen-directed immunotherapy of primary brain tumors could target cancer/testis antigens (CTAs), especially those expressed at higher frequency such as NY-ESO-1. PMID- 18396788 TI - Potential stem cell marker CD44 is constitutively expressed in permanent cell lines of head and neck cancer. AB - BACKGROUND: Despite significant advances in the use of diagnosis and therapy to treat head and neck squamous cell carcinoma (HNSCC), the prognosis has improved only marginally in the last decades. Thus, there is an enormous need for better understanding of tumor biology and reversely novel immunotherapeutic approaches. It is becoming increasingly obvious that stem cells play an important role in tumor development and progression. The identity of these cells and the underlying cellular and molecular mechanisms are mostly unknown in HNSCC to date. MATERIALS AND METHODS: Solid HNSCC tumors, as well as permanent HNSCC cell lines, were analyzed by flow cytometry concerning the expression of different putative stem cell marker proteins. RESULTS: Distinct populations of CD44 expressing potential stem cells could be identified in solid tumors of HNSCC patients with strong individual deviations. Surprisingly, the potential stem cell marker CD44 was found to be constitutively expressed on the surface of all the permanent HNSCC cell lines analyzed. CONCLUSION: CD44+ 'tumor stem cells' may play a key role in the establishment of permanent HNSCC cell lines, selecting especially robust cell entities that might drive the progression and metastasis of HNSCC. Individual analysis of 'tumor stem cell' markers will be an important tool for innovative therapies and for determining the prognosis of patients with HNSCC. PMID- 18396789 TI - Synthesis and biodistribution studies of two novel radiolabeled estrone derivatives. AB - BACKGROUND: Two 99mTc-DTPA attached estrone derivatives were synthesized and their radiopharmaceutical potential was determined using female albino Wistar rats. MATERIALS AND METHODS: Two novel radiolabeled estrone derivatives, 99mTc 2,2',2"2"'-(2,2'-(2-(3-methoxy-13-methyl-17-oxo-7, 8, 9, 11, 12, 13, 14, 15, 16, 7-decahydro-6H- cyclopenta[a]phenanthren-2-ylamino)-2- oxoethylazanediyl) bis(ethane-2,1-diyl)) bis(azanetriyl) tetraacetic acid (99mTc-2-DTPA-3-methoxy estrone) and 9mTc-2,2',2",2'"-(2,2'- (2-(3-methoxy-13-methyl-17-oxo- 7,8,9,11,12,13,14,15,16,17-decahydro- 6H-cyclopenta[a]phenanthren-4-ylamino)-2 oxoethylazanediyl) bis(ethane-2,1-diyl))bis(azanetriyl)tetraacetic acid (99mTc-4 DTPA-3-methoxy estrone) were synthesized starting from estrone (3-hydroxy-13 methyl-7,8, 9,11,12,13,15,16-octahydro-6H-cyclopenta[a]phenanthren-17(14H)-one) and DTPA anhydride (2-(bis(2-(2,6-dioxomorpholino)ethyl)amino)acetic acid) as potential estrogen receptor imaging agents. The products were crystallized in ethyl alcohol (95%), purified by high performance liquid chromatography (HPLC) and characterized by nuclear magnetic resonance (NMR) and infrared spectroscopy (IR). The effect of the radiolabeled compounds on the biological behaviour of the molecules was evaluated through biodistribution studies in female albino Wistar rats. The rats were sacrificed at various time intervals, their organs were removed, and the activities of organs were counted using a gamma counter equipped with a Cd(Te) solid state detector. RESULTS AND CONCLUSION: Organ uptake was calculated as activity/gram tissue and time versus activity curves were generated. The tissue distribution studies exhibited a receptor-mediated uptake in the target organs of the rats for each compound. Both 99mTc-2-DTPA-3-methoxy estrone and 99mTc-4-DTPA-3-methoxy estrone were stable in vitro and were mainly excreted through the hepatobiliary pathway. The biological data showed that the 99mTc-2-DTPA-3-methoxy estrone had higher uptake in the target tissues than the 99mTc-4-DTPA-3-methoxy estrone. The favourable in vitro/in vivo stability and biodistribution profiles suggest that these radioligands are good candidates for further exploration of their potential clinical applications. PMID- 18396790 TI - The frequency of lymphocytic gastritis in baboons. AB - BACKGROUND: In 1985, two independent reports highlighted a novel subtype of chronic inflammation in the gastric mucosa, characterized by the intraepithelial lymphocytic infiltration (ILI) both in the surface and the foveolar epithelium. The disease, subsequently called lymphocytic gastritis (LG) is a rare form of gastritis (0.8%-1.6% of cases), with unclear pathogenesis. More recently, LG was recorded in pigs and in non-human primates. MATERIALS AND METHODS: The frequency of LG (>25 lymphocytes/100 epithelial cells) was assessed in gastric specimens from 92 consecutive baboons, initially filed under the diagnosis of "gastritis". RESULTS: LG was found in 13 (14%) out of the 92 animals. Helicobacter pylori was not found. DISCUSSION: LG mirrors an immunological phenomenon at the surface foveolar cell level elicited by an uncertain etiological factor. In similarity to humans with LG, no Helicobacter pylori were found in baboons with LG. The search for the lymphocyte-attracting protein contained in affected cells might bring forward an alternative therapy capable of abrogating the specific surface foveolar cell-lymphotaxis present in LG. The baboon emerges as a possible animal model to study the agent(s) leading to LG. PMID- 18396791 TI - Effects of 4-tert-Octylphenol on the incubation of eggs in Japanese brown frogs (Rana japonica). AB - 4-tert-Octylphenol (OP), is an endocrine disruptor or surfactant widely used in herbicides. Its effects (0, 1 and 10 mg/l) on the incubation of eggs were examined using wild Japanese brown frogs (Rana japonica). In 10 mg/l OP, all the eggs were corrupted and no eggs developed. In 1 mg/l OP, 9.8% eggs developed and systemic edema, malformations such as crooked vertebrae and atrophy of the systemic muscles were observed in all the surviving tadpoles. These results suggested that OP use in paddy fields may affect the survival rate of wild frogs and induce malformation. PMID- 18396792 TI - Review. 18F-FDG PET in the diagnosis and follow-up of thyroid malignancy. AB - The diagnosis of carcinoma of the thyroid is usually made in the process of investigating a thyroid nodule with clinical examination, Technetium-99m scan, ultrasonography and fine-needle aspiration (FNA) cytology. The follow-up is mainly based on 123-iodine and 131-iodine scans and serum thyroglogulin measurement. The aim of the present review was to establish the role of 18F-FDG PET in the differential diagnosis of doubtful thyroid nodules and in the follow up of patients with increased serum thyroglobulin levels and negative iodine scan. It remains to be defined if metabolic imaging with PET could be a useful routine procedure in the management of thyroid tumours since the majority of them are well-differentiated and therefore have less avidity to 18F-FDG. In the present work we collected the specific literature derived from MEDLINE over the last 10 years to clarify the potential clinical value of 18F-FDG PET in thyroid malignancies. An emerging role for 18F-FDG PET is in the assessment of incidental finding of a thyroid nodule which, when showing high FDG uptake should be regarded as a possible malignancy that needs further assessment. Another well documented role for 18F-FDG PET is in the investigation of cases of established well-differentiated thyroid carcinomas presenting with high thyroglobulin and negative iodine imaging. An increase of the 18F-FDG uptake in these tumours indicates a shift towards lesser differentiation (with more aggression and poor prognosis) and may benefit from alternative management. 18F-FDG PET can be considered a routine functional imaging method in detecting iodine-negative recurrent disease in thyroid cancer patients with elevated serum thyroglobulin levels during follow-up. 18F-FDG PET seems to be useful also in differential diagnosis of suspected thyroid nodules, especially using the semi-quantitative SUV analysis. PMID- 18396793 TI - Unravelling intracellular immune dysfunctions in chronic fatigue syndrome: interactions between protein kinase R activity, RNase L cleavage and elastase activity, and their clinical relevance. AB - This study examined possible interactions between immunological abnormalities and symptoms in CFS. Sixteen CFS patients filled in a battery of questionnaires, evaluating daily functioning, and underwent venous blood sampling, in order to analyse immunological abnormalities. Ribonuclease (RNase) L cleavage was associated with RNase L activity (rs=0.570; p=0.021), protein kinase R (PKR) (rs=0.716; p=0.002) and elastase activity (rs=0.500; p=0.049). RNase L activity was related to elastase (rs=0.547; p=0.028) and PKR activity (rs=0.625; p=0.010). RNase L activity (rs=0.535; p=0.033), elastase activity (rs=0.585; p=0.017) and RNase L cleavage (rs=0.521; p=0.038) correlated with daily functioning. This study suggests that in CFS patients an increase in elastase activity and subsequent RNase L cleavage is accompanied by increased activity of both the PKR and RNase L enzymes. RNase L and elastase activity are related to daily functioning, thus evidence supporting the clinical importance of these immune dysfunctions in CFS patients was provided. PMID- 18396794 TI - Post-coronarography application of continuous veno-venous hemofiltration in the prevention of contrast nephropathy in patients with complex multisystem deficiency. AB - BACKGROUND: An increased admission of high-risk patients to diagnostic and interventional radiological procedures with contrast medium has resulted in an increase of contrast-induced nephropathy, which now represents the third main cause of hospital-acquired acute renal failure. The pathogenic mechanism of contrast-induced nephropathy (CN) is unclear, but there is much evidence which indicated an interaction between direct tubular cytotoxicity and osmotic/hemodynamic effects. Continuous veno-venous hemofiltration (CVVH) has shown possible benefits in preventing CN. It is not understood when and how prophylactic strategies should be used either in pharmacological therapies or in continous renal replacement therapy (CRRT) approaches. The aim of this study was to evaluate the efficiency of the CVVH technique in preventing CN secondary to emergency radiological procedures in very high-risk patients. PATIENTS AND METHODS: Twelve patients with severe chronic renal impairment (serum creatinine concentration >2 mg/dl with an estimated glomerular filtration rate (eGFR) <40 ml/min) in association with at least two severe comorbidities (such as previous acute myocardial infarction in hypertensive or diabetic patients obesity, cardiac failure with ejection fraction <40%, severe hypotension) were treated with CVVH after coronarography using an iso-osmolar contrast medium (Visipaque, Iodixanol), with or without percutaneous transluminal coronary angioplasty. Adverse events and their association with the interventional radiological procedure were investigated after hemofiltration. RESULTS: Statistically significant differences were observed for both eGFR and serum creatinine at different time points (pre-, post- and 7 days after the procedure) at p<0.05. Statistical analysis of all the variables related to the radiological procedure and the hemofiltration technique did not cause any modification of renal function between the pre- and post procedure values. No patient showed signs of cardiovascular instability, nor were any episodes of marked hypotension reported during the dialysis session. No patient showed any adverse effects related to the interventional radiological procedure or to the CVVH technique. Renal function, according to serum creatinine concentration and the e-GFR calculation (Cockcroft), did not worsen but had improved when the patients left hospital, with function rates statistically significantly better compared to that on hospital admission, even 7 days after the radiological procedure. CONCLUSION: The present study suggests the efficiency of the CVVH technique in preventing CN in high-risk patients who need to undergo interventional radiological cardiovascular procedures involving the administration of an iodine-based contrast medium. PMID- 18396795 TI - Factors influencing EPA+DHA levels in red blood cells in Japan. AB - The blood eicosapentaenoic and docosahexaenoic acid (EPA+DHA) concentration is an important inverse risk factor for sudden cardiac death. However, it is not known what kinds of factors influence the EPA+DHA levels in the total phospholipid fraction in red blood cells (RBC EPA+DHA) in Japan, who regularly eat more fish with increasing age. Four hundred and fifty-six healthy individuals (320 men and 136 women, 18 to 70 years old) were recruited between 2002 and 2005. RBC EPA+DHA were measured by gas chromatography and questionnaires were administered. Multivariate analysis indicated that there were significant correlations between RBC EPA+DHA and (i) dietary EPA+DHA (beta=0.31), (ii) age (beta=0.33), (iii) gender (beta=-0.15) and (iv) physical activity (beta=-0.11) but not with body mass index or smoking. PMID- 18396796 TI - Plasma glutamate and glycine levels in patients with amyotrophic lateral sclerosis. AB - Defective glutamate (Glu) metabolism and glutamate excitotoxicity have been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). Glycine (Gly), the main inhibitory neurotransmitter, has been shown to potentiate excitatory transmission. In the present study, the levels of Glu and Gly in fasting plasma were measured by high performance liquid chromatography (HPLC) in 20 healthy volunteers and in 65 untreated ALS patients. Increased plasma Glu levels were observed in ALS (p=0.05), correlating with longer disease duration (p=0.03, beta=0.34) and male gender (p=0.02). Furthermore, the increase was found only in the spinal subtype of the disease (p=0.03), while in the bulbar subtype, no significant increase was noted. As regards plasma Gly, no difference was observed between patients and controls; however female patients had higher levels than males. The above results are compatible with the "glutamate hypothesis" of ALS and suggest that the spinal and bulbar-onset subtypes of the disease may be biochemically different. PMID- 18396797 TI - Modification of the Harris-Benedict equation to predict the energy requirements of critically ill patients during mild therapeutic hypothermia. AB - The aim of the present study was the comparison of calculated and measured values of the energy expenditure of critically ill patients during the different phases of therapeutic mild hypothermia. PATIENTS AND METHODS: Five patients (mean age 40.6 years, range 23 to 68 years, 2 females) suffering from severe acute cerebral injuries who underwent mild hypothermia were prospectively included into the study. Indirect calorimetry measurements were made at intervals of 3-4 hours and subsequently, during the steady state, at least every 12 hours. The results were compared with the basal metabolic rate calculated by the Harris-Benedict equation. RESULTS: A close linear correlation between body temperature and basal metabolism could be observed across a wide range of temperatures from 30.5 degrees C to 38.3 degrees C (r=0.82, p<0.001). One degree drop in temperature led to a 5.9% reduction in energy. During mild hypothermia, oxygen consumption was reduced by 71 mL/min (95% confidence interval 57 to 86 mL/min; p<0.001) as compared to base line. The basal metabolism rate was decreased by 30.3% (95% confidence interval 24.7 to 35.9%, p<0.001). The average value recorded was 16.7% below the values calculated in accordance with the Harris-Benedict equation (95% confidence interval 12.8 to 20.6%). CONCLUSION: The immediate reduction in oxygen requirements achieved by hypothermia is linearly correlated with the reduction in temperature and the hypothermia induced reduction in oxygen requirement recorded by indirect calorimetry is considerably below that calculated in accordance with the Harris-Benedict equation. If indirect calorimetry should not be available and the Harris-Benedict equation is used, a corrective factor is therefore needed to avoid an inaccurate calorie administration. PMID- 18396798 TI - Effect of Ala16Val genetic polymorphism of MnSOD on antioxidant capacity and inflammatory response in open heart surgery. AB - BACKGROUND: Ischemia-reperfusion injury and inflammation in cardiac surgical patients involves complex humoral and cellular interactions. We investigated the effect of genetic polymorphism of manganase superoxide dismutase (MnSOD) a natural antioxidant, on cytokine release and manganesuperoxide dismutase in patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB). PATIENTS AND METHODS: Forty-two patients undergoing elective CABG with CPB were included in the study. MnSOD polymorphism was performed by polymerase chain reaction (PCR). Levels of interleukin-6 and mangane superoxide dismutase (MnSOD) were measured by enzyme linked immunoabsorbent assay (ELISA). RESULTS: Baseline IL-6 did not differ between patients with different MnSOD genotypes. Postoperatively IL-6 levels were significantly higher in all patients but more significantly in V(VV+AV) carriers (p=0.003). The wild-type AA genotype had the highest preoperative (p<0.05) and postoperative IL-6 level. The MnSOD VV genotype was associated with significantly lower preoperative MnSOD levels compared to the AA carriers (p<0.05). CONCLUSION: These data demonstrate that MnSOD Ala16Val polymorphism influences IL-6 production and baseline MnSOD activity, suggesting that preoperative MnSOD concentration plays a role in cytokine release. PMID- 18396799 TI - Multicystic peritoneal mesothelioma treated by surgical cytoreduction and hyperthermic intra-peritoneal chemotherapy (HIPEC). AB - BACKGROUND: Multicystic peritoneal mesothelioma (MPM) is an extremely uncommon lesion with uncertain malignant potential. Multiple recurrences after surgical interventions and transition to aggressive malignancies have been reported. Here, we review our experience with cytoreduction and hyperthermic intraperitoneal chemotherapy (HIPEC) in the management of MPM. PATIENTS AND METHODS: Five women with MPM underwent 6 procedures of cytoreduction and close-abdomen HIPEC with cisplatin and doxorubicin. Three patients had recurrent disease after 1, 2 and 4 previous debulkings, respectively. RESULTS: Optimal cytoreduction (residual tumor nodules < or =2.5 mm) was performed in all the procedures. One grade 4 postoperative complication (NCI/CTCAE v.3.0) and no operative mortality occurred. Median follow-up was 31 months (range 3-102). MPM recurred in two patients: one is presently disease-free after a second cytoreduction with HIPEC and the other is alive with minimal stable disease. CONCLUSION: Definitive eradication by means of cytoreduction and HIPEC seems a safe and effective therapeutic option for MPM. PMID- 18396800 TI - Susceptibility of gerbera and chrysanthemum varieties (Gerbera jamesoni and Chrysanthemum morifolium) to Fusarium oxysporum f.sp. chrysanthemi. AB - In 2002, gerbera plants (cv Kaliki) were observed exhibiting symptoms of a wilt in a soilless cultivation at Albenga area (Northern Italy). A similar wilt was also observed in the Sanremo area (Northern Italy) on cv Red Bull, Anedin and Gud finger grown in soil. The same observations were carried out in 2004 in SW Spain where gerbera plants showing wilt symptoms were observed in soilless crops. In all cases, the planting material originated from the Netherlands. Recently on the base of experimental trials F. oxysporum f. sp. chrysanthemi was recognized as the causal agent of wilts of gerbera both in Italy and in Spain. The aim of this experimental work was the evaluation of the resistance/susceptibility of available cultivars of chrysanthemum and gerbera to the Fusarium wilt. The pathogenicity of two isolates of Fusarium chrysanthemi obtained from infected gerbera (Gerbera jamesonii) and chrysanthemum (Chrysanthemum morifolium) plants was tested on several varieties both of gerbera and chrysanthemum in 2004-2006. In 2004 and 2005 respectively 54 and 30 cultivars of chrysanthemum and 57 and 55 of gerbera were tested, while in 2006 only 53 cultivars of gerbera were tested. The results showed that respectively in 2004 and 2005 67 and 33 % of chrysanthemum cultivars were highly resistant to F. chrysanthemi obtained from chrysanthemum while 57 and 53 % were highly resistant to strain isolated from gerbera. In 2004, 2005 and 2006 47, 65 and 75 % of gerbera cultivars were highly resistant to F. chrysanthemi obtained from chrysanthemum and 48, 56 and 72 % were highly resistant to the strain isolated from gerbera. PMID- 18396801 TI - Yield loss in susceptible cultivars of spring rapeseed due to Fusarium wilt caused by Fusarium oxysporum. AB - In 1999, reports of spring rapeseed plants (Brassica napus L.) exhibiting wilt symptoms were received by agricultural extension personnel from farmers near Fort Vermillion and Andrew, Alberta, Canada. Fungal colonies recovered from affected plants after surface disinfection were identified as Fusarium oxysporum by comparison of morphology on carnation leaf and potato dextrose agars with literature descriptions and reference cultures. Root-dip inoculation of young rapeseed plants with spore suspensions prepared from recovered F. oxysporum colonies resulted in rapid development of symptoms seen in the field. An initial estimate of yield loss in an affected field near Andrew was performed by removing all rapeseed plants from three 1 m2 quadrats. Each plant was evaluated according to a simple three point severity scale, and then the seed from each plant was individually threshed and weighed. Fully- and partially-wilted plants yielded 0.2 and 19.3% of asymptomatic plants, respectively. In 2000, wilt symptoms were observed at a plot research site near Ranfurly, Alberta. Disease symptoms were restricted to one B. napus cultivar, Nexera 705. A similar procedure to that used at Andrew in 1999 was applied at Ranfurly, except quadrat size was 2.5 m2, and replicated comparisons were made between Nexera 705 and an unaffected cultivar, Quantum. The average number of unaffected Quantum plants was 99.4%, while only 66.9% of Nexera 705 plants were asymptomatic. No Quantum plants were severely wilted, while 11.7% of Nexera 705 plants were wilted. Yield of Nexera 705 was 38.6% of Quantum. In 2004, the impact of fusarium wilt on yield of 6 susceptible and 9 resistant B. napus cultivars was determined at nine locations in western Canada. Cultivars were selected on the basis of survey results and agronomist's reports. Across all sites, yield of the most severely affected cultivar, 45A55, was 15.9% lower than the least severely affected cultivar 3455. At the most severely affected site, yield of the most severely affected cultivar, Bianca II, was 75.2% lower than the least severely affected cultivar, Option 501. Yield of wilt-susceptible (Canterra 1604) and resistant (Cougar CL) rapeseed was compared at a replicated large-plot (576 m2 per plot) experiment at Lavoy, Alberta in 2005. The susceptible variety was severely affected and yielded 44% of Cougar CL. PMID- 18396802 TI - First occurrence of strobilurin-resistant isolates of Stemphylium vesicarium in an Italian pear orchard. PMID- 18396803 TI - Molecular detection of Puccinia horiana Henn. the causal agent of Chrysanthemum white rust. AB - Chrysanthemum white rust is one of the most important foliar diseases of pot chrysanthemum and is a quarantine pathogen in many countries. Under conducive environmental conditions, it has the potential to completely destroy susceptible cultivars. This is mainly avoided through frequent preventive fungicide applications. As part of a research program to develop a disease warning system, a molecular detection method was developed. To determine the nucleotide sequence of the nuclear rDNA-ITS (internal transcribed spacer) region of P. horiana, 56 isolates were collected between 2003 and 2006 from diseased commercial chrysanthemum plants from different national and international geographical areas. DNA was isolated from the basidiospores or teliospores from several isolates and the rDNA-ITS region was cloned and sequenced. Based on the limited variability in rDNA-ITS sequence between these isolates, several primer pairs were designed and tested for detection through conventional and real-time PCR. Specificity of detection was cross-checked against a variety of other fungi (saprophytes and other rusts) that may occur in the same environment, and against DNA of healthy chrysanthemum leaves. Using the best primers, the PCR-based methods successfully detected all the P. horiana isolates tested, while no signal was observed with other rust species up to 1 ng non target genomic DNA template. The limit of detection of P. horiana DNA in conventional, nested and real-time PCR was 10 pg, 10 fg and 10 fg, respectively. The DNA extraction method and PCR template concentration were optimized to maximize the recoverability of the pathogen from infected plant tissue. Using the optimized real-time PCR method, the pathogen could be detected in washed plant tissue, 9 hours after inoculation. Hence, this method allows detection of the P. horiana in any part of its latent stage and will also serve as a tool for studying the biology and epidemiology of the pathogen. PMID- 18396804 TI - A certain but non-exclusive association between Polymyxa graminis special forms and cereals. AB - Polymyxo graminis, a ubiquitous plasmodiophorid obligate root endoparasite, is recognized as the vector of about 15 viruses on cereals and groundnut in temperate and tropical areas. Within the species, five special forms have been distinguished on the basis of specific ribotypes. Three of them occur in tropical areas: P. graminis f.sp. colombiana on rice, P. graminis f.sp. subtropicalis on cereals cropped in the tropics such as maize, pearl millet and sorghum but also on barley and/or wheat, and P. graminis f.sp. tropicalis mainly on maize, pearl millet and sorghum. Their particular host ranges distinguish them significantly from P. graminis f.sp. temperata and P. graminis f.sp. tepida found in temperate areas on barley and wheat. In order to assess whether these special forms commonly infect these cereals, barley and wheat plants were grown under controlled conditions on two soils from Belgium and France and both infested by P. graminis f.sp. temperata and P. graminis f.sp. tepida. The infection of each cereal species by each form was quantified by real-time quantitative PCR with specific primers and Taqman probes. The infection of P. graminis f.sp. temperata was significantly higher on barley than on wheat, whereas the quantities of P. graminis f.sp. tepida on wheat were higher than on barley. These results show that the distinction between these special forms, based on the ribotype, reflects differences in ecological features. PMID- 18396805 TI - Soft-rot resistance test of in vitro preselected stress tolerant potato genotypes. PMID- 18396806 TI - Naz, a resistant cultivar on bean root rot disease in Zanjan province, northwest Iran. AB - Field bean is a major crops in different parts of northwest Iran especially Zanjan province. Recently the bean plants were severely subjected to damping off or decline disease which caused yield losses in bean growing regions. A regional research was done from 2003 to 2005 to get general information on the causal agent of disease and its control management. Infected plants were collected from different studied areas and transferred to laboratory. Crown and plant roots were cultured in PDA as common media and PPA as selective media for Fusarium species after surface sterilization with sodium hypochlorite. Plates were incubated in standard culture room then isolated fungi were identified. Different Fusarium species were isolated, however the main pathogen isolated from plant samples and soil around the roots was F. sambucium Fuckel. The disease caused up to 50% yield losses in some fields in studied areas. Study showed the "Naz" cultivar was the main resistant race to the disease and had the most yield production in the field. PMID- 18396807 TI - Phytopatological problems and solutions in the walnut orchards along Lake Balaton. AB - European or Persian Walnut (Juglans regia) is an important and healthy food as well as base material of timber industry. Several pests (pathogens and insect pests) may cause serious damages on walnut. These are less known on the crop land of the tree. Results of some years of our experiments including bacteriological and mycological studies, are presented in this paper. The optimum time of chemical protection against the walnut blight (Xanthomonas arboricola pv. juglandis) was determined. Occurrences of pathogenic fungi were surveyed in an orchard and on home garden trees in Hungary (18 fungus species were identified). The following experimental results are reported on the pathogenic fungi: cultivar resistance to walnut anthracnose (Gnomonia leptostyla), dying of wood parts in the cultivar collection, application of the spore trap, in vitro fungicide testing against Phomopsis juglandina. PMID- 18396808 TI - Red stele disease of strawberry in the Philippines. PMID- 18396809 TI - Evaluation of antifungal activity of carbonate and bicarbonate salts alone or in combination with biocontrol agents in control of citrus green mold. AB - The aim of this research was to determine if the attacks of green mold on orange could be reduced by edible salts alone or in combination with biocontrol agent. For this purpose toxicity to Pantoea digitatum and practical use of sodium carbonate (SC), sodium bicarbonate (SBC) and potassium carbonate, and potassium bicarbonate alone or in combination with antagonistic bacteria (Pseudomonas fluorescens isolate PN, Bacillus subtilis isolate VHN, Pantoea agglomerans isolate CA) to control green mold were determined. All were fungistatic. SC and SBC were equal and superior to the other salts for control of green mold on oranges inoculated 6h before treatment and were chosen for subsequent trails under cold storage conditions. The biocontrol agents were found completely tolerant to 3% sodium bicarbonate and sodium carbonate at room temperature; although their culturability was reduced by > 1000-fold after 60 min in 1% other salt solutions. Satisfactory results were also obtained with the combined treatment for control of green mold. A significant increase in biocontrol activity of all isolate was observed when combined with sodium carbonate and sodium bicarbonate. The treatments comprising CA combined with SB was as effective as fungicide treatment. Thus, use of sodium bicarbonate treatment at 3% followed by the antagonist P. agglomerans CA could be an alternative to chemical fungicides for control of green mold on oranges. PMID- 18396810 TI - Comparison of different techniques for inoculation of "Candidatus Phytoplasma mali" on apple and periwinkle in biological indexing procedure. AB - As phytoplasmas are non cultivable micro-organisms, the research on phytoplasmal diseases can only be achieved with infected hosts. Biological indexing (by grafting) is the simplest detection method for phytoplasmal diseases. We tested four different grafting techniques for inoculation of apple trees or periwinkles in greenhouse, including whip graft, bark graft, budding and chip-budding. All techniques were tested on apple trees (six trees per phytoplasma isolates) in insect-proof greenhouse. The whip and bark grafting were not feasible for periwinkle plants, because of fineness and fragility of their tissues: only the chip-budding was performed (four plants per isolate). In apple trees, the best and soonest positive results were obtained by chip and bark grafting. Except for seven transplants not-grown after grafting, 100% efficiency of inoculation was obtained by both methods. Nevertheless, the transmission of phytoplasma from transplant not-grown to rootstock was sometimes recorded (28.6%). The earliest phytoplasma symptoms after whip or bark grafting appeared after 3 months. Symptoms were obtained much later with budding and chip-budding. In case of periwinkles, infected apple and periwinkle materials were used as inoculum sources. Transmission of phytoplasma from periwinkle to periwinkle was successfully carried out by chip-budding grafting. The symptoms were observed during the second month after inoculation. The transmission of phytoplasma from infected apple material to periwinkle (by chip-budding) was achieved for 60 % of the tested samples. Moreover, the latency period before symptom observation was longer. Finally, we perceived the apple trees are more convenient and rapid than periwinkle plants for biological indexing of apple materials. PMID- 18396811 TI - Evaluation of coumarin derivatives as anti-fungal agents against soil-borne fungal pathogens. AB - Development of new and safer pesticides that are target-specific is backed by a strong Federal, public and commercial mandate. In order to generate a new generation of pesticides that are more ecologically friendly and safe, natural products are being evaluated for pesticidal activities. Many plant-derived chemicals have proven pesticidal properties, including compounds like sesamol (3,4-Methylenedioxyphenol), a lipid from sesame oil and coumarins (1,2 Benzopyrone) found in a variety of plants such as clover, sweet woodruff and grasses. Both of these plant-derived compounds have been shown to inhibit a range of fungi and bacteria and it is believed that these cyclic compounds behave as natural pesticidal defense molecules for plants. These compounds represent a starting point for the exploration of new derivative compounds possessing a range of antifungal activity and for use as seed protectants. Within this study, six derivatives of coumarin that resembled sesamol's structure were screened for their antifungal activity against a range of soil-bome plant pathogenic fungi. Fungi in this in vitro screen included Macrophomina phaseolina (causal agent of charcoal rot) and Pythium spp. (causal agent of seedling blight), two phylogenetically diverse and economically important plant pathogens. Preliminary studies indicate that many of these novel coumarin derivatives work very effectively in vitro to inhibit fungal growth and several coumarin derivatives have higher antifungal activity and stability as compared to either the original coumarin or sesamol compounds alone. Interestingly, several of these highly active coumarin derivatives are halogenated compounds with solubility in water, and they are relatively easy and inexpensive to synthesize. These halogenated coumarin derivatives remained active for extended periods of time displaying 100% inhibition of fungal growth for greater than 3 weeks in vitro. In addition to the in vitro fungal inhibition assays, preliminary phytotoxicity assays of these halogenated coumarin compounds show no obvious plant toxicity issues or interference in plant development. These results support additional research in this area of natural pesticide development. PMID- 18396812 TI - Evaluation of the effects of chemical versus biological control on Botrytis cinerea agent of gray mould disease of strawberry. AB - This study investigates on effects of four fungicide and six isolate from Trichoderma and Gliocladium on Botrytis cinerea agent grey mold of strawberry under library and greenhouse condition. The effect of four fungicides i.e. benomyl, dichlofluanid, captan and triadimenol on B. cinerea was studied in the laboratory condition by method mixed poison to culture medium. It was shown that the fungicide including benomyl, triadimenol, dichlofluanid and captan were able to inhibit mycelial growth of B. cinerea on PDA plate with EC50 of 0.16, 1.42, 3.40 and 7.73 ppm respectively. These fungicides delayed myceliogenic germination of sclerotia at 1000 ppm, while exhibiting no fungicidal effect. Moreover, the antagonistic effects of six fungi including Trichoderma koningii (T21), T. viride (T4), T. harzionum (T5), T. viride (T2), G. virens (G2), G. virens (G8) on B. cinerea were assessed. This assessment was done under library condition and its results as follows: The antagonistic mechanism occurred through branching at the end of B. cinerea hyphae, hyphal contact, coiling, vacuolization and lyses. Volatile metabolites of T. koningii (T21) and non-volatile metabolites of G. virens (G2 and G8) and T. koningii (T21) caused maximum inhibition of the fungal growth. Trichoderma spp and G. virens were able to colonize and sporulate on sclerotia and caused their lysis within 7-21 days. In greenhouse, a completely randomized design with 11 treatments (4 chemical and 6 biological and one untreated control) each replicated five times were used for the comparison. Greenhouse studies revealed that application of fungicides i.e. captan, dichlofluanid, triadimenol and benomyl reduces disease severity by 42, 45, 48 and 52% respectively. The fungal antagonists reduce the grey mold disease severity between 5-42%. All treatments caused a decline in post harvest disease, as the most effective treatment of chemical control was benomyl with 68.33% and for the biological treatment this was T. koningii (T21) with 56%. PMID- 18396813 TI - Occurrence of downy mildews on ornamental plants and their control by chemical compounds. AB - The downy mildew on Coreopsis grandiflora caused by Plasmopara halstedii was observed during summer, mainly in July and August. Symptoms of the disease were first seen on external leaves and progressively spread to inner parts of plant rosette. On Alyssum saxatile downy mildew symptoms induced by Peronospora parasitica were observed during whole vegetation period with the strongest expression in early spring and late summer. Amistar 250 SC (25% azoxystrobine), Mildex 711,9 WG (66.7% phosethyl aluminium + 4.4% fenamidone), Previcur Energy 840 SL (530 g/l propamocarb + 310 g/l phosetyl aluminium) and Tanos 50 WG (25% cymoxanil + 25% famoxate) were used for pathogens control. In the protection of Coreopsis grandiflora against P. halstedii all tested compounds, applied curatively, decreased sporulation of the pathogen. On treaded plants at least 4 time less leaves were diseased. In the control of P. parasitica on Alyssum saxatile, the smallest number of swallowed structures on leaves was noticed on plants treated with azoxystrobine at conc. 250 microg/cm3. PMID- 18396814 TI - Introduction a potato cultivar "sprit" as relatively resistant to main fungal pathogens causal agents of early blight and wilting on potato in Iran. AB - Potato (Solanum tubersum L.) is one of the most human food production cultured in Iran especially Zanjan province as a temperate region. Some fungal pathogens caused severely infected on potato tubers or foliage in the majority grown areas and resulted yield losses in potato production. Recent years from 2002 to 2004 infected samples were collected from different potato grown regions in Zanjan province then cultured on PDA after surface sterilization with sodium hypochlorite. Isolated fungal pathogens were identified and study showed the main pathogens with high incidence and frequency were Alternaria solani, Fusarium oxysporum and Verticillium sp. in studied areas. The regions which used convention varieties showed more diseases than other locations which used relatively resistant races. The rate of resistance for 10 international potato varieties was studied by inoculation of them by 10(5) spores suspension of three common fungal pathogens in the field. Study showed Sprit cultivar was more resistant than others to all three common pathogens and Lady-Claire was most susceptible. Yield production of Sprit per unit of land area was also exceeded that of other cultivars by factors of 1.10 to 2.25 respectively. The results of the study helped potato growers to culture Sprit cultivar and have good yield production in Zanjan and Hamedan provinces in this year. PMID- 18396815 TI - Examination of resistance of potato genotypes to Erwinia ssp. AB - Potato can be attacked by several economically important pathogens. From the various diseases, in our experiment we dealt with the bacterial soft rot of potato caused by Erwinia species. In the experiments back cross progenies (BC1, BC2, BC3 and BC4) of Solanum brevidens + Solanum tuberosum somatic hybrids produced by the Potato Research Centre, Keszthely were tested to the infection of E carotovora ssp. atroseptica (Eca), E. carotovora ssp. carotovora (Ecc) and E. chrysanthemi (Echr). All together 11 BC genotypes pre selected from several hundred breeding lines based on their preferred agronomical appearance and virus resistance characters as well as 4 Hungarian potato cultivar (Rioja, Desiree, White lady and Hopehely) as controls were involved into the experiments. Tuber slices from each genotype were artificially infected with bacteria suspension Ecc strain D3, and Echr strain CHR 1492, and Eca strain BN3) and incubated at 27 degrees C with 100% relative air humidity for 48 h before evaluation. Dry matter and starch content of tubers were determined right before the tests. Volume of rotted tuber tissue was determined in mm3 and used for comparison of the level of resistance or susceptibility of the genotypes. Relationship between the reaction to the bacteria strains and dry matter content was examined also. Tested genotypes showed the highest resistance to Eca, while the highest susceptibility to Echr. By the increase of BC level the susceptibility of the genotypes significantly increased as well regardless of the tested bacteria. No direct correlation was found between the dry matter content of tubers and their reaction to tested bacteria. PMID- 18396816 TI - Effectiveness of some substances in the control of carrot and parsley roots against fungal diseases. AB - Field experiments were carried out in the years 2005 and 2006 on carrot cv. 'Koral' and 'Perfekcja', and parsley cv. 'Berlinska' and 'Cukrowa'. Effectiveness of substances: Biochikol 020 PC (biologically active substances BAS--chitosan 20 g/dm3), Bioczos BR (extract of garlic 10 g/1 brick) and Biosept 33 SL (extract of grapefruit 33%) on seedling roots of carrot and parsley was studied. As the standard fungicide Zaprawa Funaben T (carbendazim 20% + tiuram 45%) was used. Roots of carrot and parsley were treated one of tested substances spring immediately before planting seedling roots. During vegetation period the growth of seedling shoots and setting of seeds, and their infestation by fungal and bacterial pathogens was noticed. Among substances used for spring dressing of carrot and also parsley seedling roots, the best efficacy exhibited Zaprawa Funaben T in both years of observation. The highest yield of carrot seeds had combination roots cv. 'Koral' and parsley seeds roots cvs 'Berlinska' and 'Cukrowa' dressed Zaprawa Funaben T. Effectiveness of biopreparates Biochikol and Biosept was lower in comparison with the standard fungicide, but their protective effect was significantly higher than in control. Bioczos had the lowest control efficacy. PMID- 18396817 TI - Differential interaction between melon cultivars and race 1.2 of Fusarium oxysporum f.sp. melonis. AB - One hundred and ten melon accessions from different origins were evaluated for resistance to race 1.2 of Fusarium oxysporum f.sp. melonis (Fom). Artificial inoculations were performed and plants were scored for presence or absence of symptoms. The screening revealed that sources of resistance to this fungus are limited. However, a few resistant accessions were found. The accessions 'Kogane Nashi Makuwa', 'C-211' and 'C-40' from Japan provided the highest resistance to both patotypes of race 1.2 of Fom which seems to confirm that the Far East is a rich area in resistances to this race. We also detect an interesting degree of resistance in 'BG-5384', a Portuguese accession that may be classified as belonging to the botanical variety inodorus. PMID- 18396818 TI - Wilting of date palm branches by Fusarium oxysporum in south of Iran and its control managements with soil solarization method. AB - Wilting of some branches in nurseries and orchards of date palm were studied in south of Iran including Ahvaz and Abadan cities in 2005-2006 years. Different infected plants were visited and samples showing symptoms including wilting or death of branches collected from various areas and transferred to laboratory. Samples were cultured in common media (PDA) and different fungi were studied and identified. The most frequently isolated pathogen was Fusarium oxysporum which caused wilting of some branches of date palm seedling or trees in studied areas. Results showed that the disease caused main losses where date palm cuttings were cultured in infected soils, previously cropped to susceptible plants. Since chemical control was not managed the disease, soil disinfestations by soil solarization method was carried in Ahvaz as the warmer climate in studied areas to control the pathogen. Application of this method reduced population density of the pathogen from 1800 CFU -g/soil to 600 after 5 week. This method was simple, effective, non negative side and economic which can be used in nearly warm areas. PMID- 18396819 TI - Effect of different temperature and culture media on the growth of Macrophomina phaseolina. AB - The charcoal root disease caused by Macrophomina phaseolina (Tassi) Goidanich may cause considerable damages in hot as well as in dry seasons. The effect of temperature and culture media were studied on the growing patterns of 35 M. phaseolina isolates, collected from different districts of Hungary. The isolates were grown at 10, 15, 20, 25, 30, 35 and 40 degrees C temperatures respectively, and additionally at 25 degrees C on potato-dextrose-, malt-extract-, Czapek-Dox-, Sabouraud-glucose-, maize-flour- and watery agar media, using 90 mm Petri-dishes, 4 repetitions in each case. For all the isolates the most favourable temperature regime was 25 to 35 degrees C and the most advantageous media was the malt extract-, Sabouraud-glucose- and potato-dextrose-agar media. At these conditions (temperatures and culture media) mycelia growth and the diameter of microsclerotial colonies reached the 90 mm at the 5th day. Mycelia growth of the pathogen was very low at 10, 15 and 40 degrees C, and did not form microsclerotia. On watery agar microsclerotial colony seldom developed, it needed 14 days, and no continuous mycelia developed even in a 8th months culture. Diameter of microsclerotia measured on different culture media varied between 39 308 microm. PMID- 18396820 TI - Survey of microfungi on Alnus glutinosa (L.) Gaertn. from Munsterland, Germany. AB - During our investigation on microfungi in Antoniusheim, Fleissenbach und Merfelderbruch near Dulmen in Munsterland in the years 2005 and 2006 we were able to collect and identify 25 microfungi on Alnus glutinosa (L.) GAERTN. Among them are some which are very rare in Germany linke Phragmoporthe conformis (Berkley & Broome) Petrak, Cryptosporiopsis alnea (Rostr.) Petr., Prosthecium auctum (Berk. & Broome) Petr., Taphrina alni-incanae (Kun.) Magn. [= T. Amentorum (Sadeback) Rostrup], Cryptodiaporthe oxystoma (Rehm.) Z. Urb., Cladosporium alnicola Bub. & Vleug. [= C. Herbarum (Pers.)], Erysiphe penicillata (Wallr.) Link, Melampsoridium betulinum Kleb., Bacterodesmium longisporum M.B. Ellis, Marssonina alni Karak. Asteroma alneum (Pers.: Fr.) Sutton . All collected species can be found in the text. PMID- 18396821 TI - Remarkable microfungi from Oaxaca (Mexico) of Acacia species. AB - In the state of Oaxaca (Mexico, 10 km north-west of Puerto Escondido 15 degrees 55' N, 97 degrees 09' W) we were able to collect some microfungi living as parasites or saprophytes on Acacia species, some of them are causing attention for Oaxaca. Many belong to the Deuteromycotina (Hyphomycetes, Coelomycetes) and Ascomycotina. On A. hindsii: Calonectria pseudopeziza (Desm.) Sacc., Hypoxylon truncatum (Schwein. Fr.) J.H. Miller, Epicoccum nigrum Link., Zygosporium gibbum (Sacc., M. Roussau & E. Bommer) S.J. Hughes and on A. cornigera: Phyllosticta acaciicola P. Henn., Taeniolella alta (Ehrenb. ex Pers.) S.J. Hughes, Cephaliophora tropica Thaxt., Diplodia mutila (Fr. Fr.) Mont., Pleospora herbarum (Pers. Fr.) Rabenh., Gliocladium roseum Bainier, Ulocladium atrum Preuss., and different others. All species collected are listed in text. PMID- 18396822 TI - Two Mycosphaerella graminicola French isolates differ in symptoms, in planta sporulation and cell wall degrading enzymes in vitro production. AB - The pathogenicity level of two French Mycosphaerella grominicola field isolates (T0414 and T0251) was evaluated on Soissons wheat cultivar using two methods: detached wheat leafs assay in a climatic chamber and wheat seedlings assay in a greenhouse. Both methods revealed that chlorosis and necrosis caused by the T0414 isolate were larger than those caused by the T0251 isolate. Indeed, in the detached leaf assay, the first symptoms on leaves inoculated by T0414 were observed 12 days post inoculation and 50% of the leaves were infected on day 18, with a maximum of 84% of leaves on day 22. On the other hand, the first symptoms were observed on day 18 on leaves inoculated with T0251 with a maximum of only 10% of leaves that were infected on day 22. In addition, both methods showed that necrosis surfaces caused by T0414 were covered by large number of pycnidia, while no pycnidia were observed on the reduced necrosis caused by the T0251 isolate. To understand the pathogenicity variation between these two isolates, their ability to produce cell wall degrading enzymes, xylanases and polygalacturonases was investigated in vitro every 2 days for 20 days. The results showed similar time course for production of polygalacturonases for the two isolates, with non significant higher production for T0414. However, a peak of maximum production of xylanases by T0414 (343 +/- 52 mU mL(-1)) was observed on day 12 post inoculation, while the maximum production (265 +/- 72 mU mL(-1)) by T0251 was observed only on day 20 post inoculation. This result shows a relationship between the beginning of the appearance of symptoms on detached leaves inoculated by T0414 on day 12 and the maximum of xylanases production on the same day in enzymes assays. In conclusion, this study suggests pathogenicity variability between M. graminicola isolates and the role of xylanases in the pathogenicity of this fungus. PMID- 18396823 TI - Phoma glomerata as causal agent of crown rot disease of fennel in southern Italy. AB - In the autumn seasons of the years 2005 and 2006 millions of fennel seedlings under greenhouse in nurseries where observed in Basilicata, Calabria, Campania and Apulia Regions, southern Italy, with serious symptoms of damping off and root necrosis. The basal part of seedling stem was much thinner than the uninvaded parts above it and the crown was brown and shrink. The incidence of disease reached values ut to 100% in some blocks where the humidity was higher, and seedlings attached earlier died. Almost all fennel varieties were interested by the disease, some of them more seriously. Seedlings attached later recovered to a great extent after transplanting in open field. In this work the identification of the causal agent of this disease was made applying Koch's postulates. Moreover, for a rapid and unambiguous identification of the fungus species, the internal transcribed spacers and the 5.8 rDNA gene (ITS1-5.8-ITS2) were amplified from DNA extracted from the isolated colonies. The fragment of amplified DNA was sequenced and compared with those present in Gene Bank (NCBI). The fungus isolated after the morphological and molecular characterization was ascribed to the species Phoma glomerata (Corda) Wollenweber & Hochapfel. This is the first report of P. glomerata as agent of crown rot of fennel. PMID- 18396824 TI - Involvement of Phytophthora species in the decline of beech Fagus sylvatica in Wallonia (Belgium). AB - During the last decade, typical symptoms of Phytophthora diseases were observed in beech stands of several European countries. The main symptoms were the presence of bleeding cankers on the stem, a low crown density as well as the yellowing of foliage and the small size of leaves. Several species of Phytophthora, such as Phytophthora citricola, P. cambivora and P. cactorum, were reported as the causal agents. In order to evaluate the implication of the different Phytophthora species in beech decline in the southern part of Belgium (Wallonia), a monitoring was undertaken with the help of managers of public and private forests. Phytophthora strains isolated from beech of different stands as well as from soil were characterized through morphological and molecular analyses (PCR-RFLP of ITS). All the isolated strains were identified as P. cambivora, except for one strain whose identification is ongoing. Molecular analysis was also directly applied to necrosed tissues of bleeding beeches and enabled the detection of additional cases. All positive cases exhibited a profile characteristic of the species P. cambivora, except for one of the sampled trees showing a different Phytophthora profile also corresponding to the unidentified isolated strain. Identification of the Phytophthora species linked to this different RFLP profile is also ongoing. Both complementation types (A1 and A2) of P. cambivora were identified, sometimes in the same sampling site. Ornamented oogonia characteristic of this species were produced by pairing A1 and A2 strains isolated from the same site. PMID- 18396825 TI - Phytophthora spp. associated with nursery reservoirs, rivers and drainage canals. AB - Phytophthora spp. were recovered from water using rhododendron leaves as baits for detection of that group of organisms. They were was found in 4 rivers, 2 hardy nursery water reservoirs and nursery drainage canal from May to October, 2006. Analysis of spots number on rhododendron leaf baits as the measure of Phytophthora spp. density showed that place of baits holding had significant influence on the species occurrence. Significantly more spots, especially in July surveying, were observed on baits holding in Skierniewka and Zwierzynka rivers swimming through agriculture and forest area than in Ner, the river of horticulture area. In nursery water containers and drainage canal higher Phytophthora density was noticed on August than other periods of surveying. PMID- 18396826 TI - Biocontrol of Phytophthora cactorum the causal agent of root and crown rot on apple (Malus domestica) by formulated Pseudomonas fluorescens. AB - In this study 284 isolates were isolated of apple trees' rhizosphere from Iran and 128 isolates were obtained from the collection of Research Department of Biological Control of University of Tehran. Four strains (P60, P61, P96, and P97) of Pseudomonas fluorescens were selected for greenhouse trials. The results of greenhouse trials showed dipping the crown and root of apple seedlings (MM106) combined with soil drench was more effective than dipping the crown and root on reducing the disease. After 6 weeks, strain P60 in dipping method combined with soil drench with 70% control, exhibited greatest effect on reducing the crown and root rot and was more effective than the fungicide metalaxyl-mancozeb. After 12 weeks, strains P60 and P96 in dipping method combined with soil drench with 55.6% and 44.5% control respectively, exhibited greatest effects on reducing the diseases Study of media on growth rate populations of effective strains exhibited that the beet molasses yeast extract (1:1) had more effect than nutrient broth(NB) medium. The initial high populations of powder formulations of strains P60 and P96 decreased during the storage at 4 and 25 degrees C over a 150-day period. In addition, formulations of strains stored at 4 degrees C had longer shelf life than those stored at 25 degrees C. In glasshouse trials, after 6 weeks, formulation of strain P60 and unformulated P60, obtained from NB medium and formulated P60, obtained from molasses yeast extract medium, and metalaxyl mancozeb had highest effect on reducing the disease on apple rootstocks. After 12 weeks, formulation of strain P60 and unformulated bacteria obtained from both media, and metalaxyl-mancozeb with 57.1% control showed greatest effect on reducing the disease. PMID- 18396827 TI - More years of field trials against Plasmopara viticola in organic viticolture. AB - The organic viticulture assumes a decisive role in the national agricultural sector. More impelling problems in the management of organic vineyards are represented from the plants pathology defence and particularly of Plasmopara viticola containment. Copper represents one of the few usable fungicides in the organic farming and the only effective against downy mildew. With Regulation EC n. 473/2002, fixed maximum quantity usable of copper compounds, owing to the environment problems due to the copper accumulation in the soil. To reduce quantity of metal copper or replace it with natural products, are conducted field trials with copper compounds at a low rate or alternative to copper products. Besides, we are estimating possibility to reduce the operations against P. viticola optimizing fungicidal treatments. Field trials in the organic farms located near Rome, have been carried out. Guidelines EPPO/OEPP PP 1/31 (3) have been carried out. The results of the trials have showed that, using cupric products with low metallic content, to reduce copper quantities used, always allowed to respect the limits established by Regulation EC. The alternative products that were investigated have not guaranteed, instead, an adequate protection in high pressure of grapevine downy mildew. It was possible to reduce treatments against P. viticola through control of different environmental parameters. The trials confirm that the copper is indispensable for plant protection in organic farming as it is not possible to replace it with natural extracts substances. We can reduce, instead, the copper quantities used trough the use of new products with low quantity metal copper or through the evaluation of climatic and pedologic data that allow to rationalize the fungicidal treatments. PMID- 18396828 TI - Screening preharvest/postharvest strategies to prevent fruit rot decay. AB - In fruit growing preharvest sprayings in the orchard are mainly applied to protect fruit from decaying. Next to multisite fungicides (captan, thiram, tolylfluanid) the most commonly used products recognized for the Belgium market are Bellis (pyraclostrobin & boscalid) and the combination of Topsin M (thiophanate-methyl) and Frugico (diethofencarb). In general the spraying schedule varies depending on weather conditions (infection risk), preharvest interval of available fungicides, fruitgrower and cultivar of pome fruit (apple/pear). Facing the climatological conditions before picking the residue loading on the fruit surface can differ enormously. Also wet (pre)grading is considered to decrease the product residue resulting to fruits which are less protected before entering the cold storage room. In this context a partially replacement of the preharvest treatments by one postharvest application could offer a reliable alternative to the PPP reduction program (Plant Protection Products) in the orchard. A standardized application method by dipping or drenching will cover the fruits homogenically resulting in a rationalized fungicide use compared to the preharvest sprayings in the orchard. For the Belgium market Philabuster (imazalil & pyrimethanil) is registered for postharvest treatments since for this product a proper solution for the waste water of postharvest uses was developed to protect surface waters (Funds technology). Philabuster provides an advanced mould control towards fruit rot pathogens Gloeosporium spp., Botrytis cinerea and Penicillium spp. In this context several trials were set up to evaluate the biological efficacy of Philabuster alone or in combination with preharvest sprayings in the orchard. In concrete different preharvest spraying schedules were applied in the last six weeks before harvest on apple and pear facing parameters as rational fungicide use, antifungal effectiveness and cost price. The purpose was to select the optimal combination in use of preharvest fungicides with Philabuster as postharvest treatment, which offer full protection towards all key pathogens in apple and pear. PMID- 18396829 TI - Evaluation of the users value of salts against apple scab and powdery mildew for the integrated fruit production. AB - As new fungicides are mainly unisite action fungicides, the problem of fungicide resistance development is becoming more important every year. Combining chemical fungicides, which is the best anti-resistance strategy, is not always possible or recommended in the case when the number of available chemical fungicides are limited or a reduction in fungicide use is asked for. Therefore the use of salts as an anti-resistance strategy was looked upon. The salts evaluated were K(HCO3), KH2PO3, KHPO4 and K2SiO3. When using these salts as an anti-resistance strategy the efficacy obtained when spraying the compounds alone was often to low to be used in rotation with chemical fungicides. Only with K(HCO3)2 a good efficacy can be observed in some years. The variation in efficacy with K(HCO3)2 observed is higher for powdery mildew. Chitosan was also included in the trials against powdery mildew, however chitosan had no effect on the infestation. PMID- 18396830 TI - A new report of pre-harvest ear rot of corn caused by Geotrichum candidum from Iran. AB - In summer of 2004, samples of husk looseness ear of corn (Zea mays) (cv. 700 Karaj) were collected from corn fields in Ali-Abad (Jiroft region), Kerman province, Southeastern Iran, for diagnosis of an unusual ear decay. A fungus was isolated from the rotting kernels and subsequently identified as Geotrichum candidum. The fungal pathogen was found to be closely related to G. citri aurantii (citrus race) based on morphological, physiological and pathogenicity properties. The fungal pathogenicity test was demonstrated by fulfilling Koch's postulates. The pathogen caused rot disease on husk looseness corn kernels in soft-dough stage of ripening. The fungus was also pathogenic on ripe lemon and green and ripe tomato fruits. Fungal isolates of corn were compared to isolates from soft-rotten potato tubers. These two groups of isolates were highly similar on the basis of their morphological, biochemical and pathogenicity characteristics. To our knowledge, this is the first known report of corn ear rot caused by G. candidum in the world. PMID- 18396831 TI - Evaluation of production of some antifungal metabolites by fluorescent pseudomonads in vitro and their inhibitory effect on Penicillium digitatum in semi-commercial condition. AB - Epiphytic fluorescent Pseudomonads isolated from the surfaces of citrus leaves and fruits collected from the citrus orchards located in the north of Iran were screened for antagonistic activity against Penicillium digitatum caused agent of green mold of citrus fruit. 9 isolates revealed antagonistic effect in dual culture and then evaluated for production of antimicrobial metabolites such as production hydrogen cyanide on King's B medium containing glycine, production of siderophore on blue CAS-agar medium and production protease enzyme on Skim Milk Agar (SMA) medium. About 28% strains produced hydrogen cyanide, 83% strains produced siderophore and 70% of them produced protease. Antagonistic effects of the screened 9 isolates were studied by inoculating of the infected orange fruits with the bacterium at the concentration of 10(8) cells/ml under sterilized condition at 20 degrees C. The three final screened antagonist isolates were more studied in semi commercial trails in cold storage with dipping of the orange fruits in the bacterial suspension at the concentration of 10(8) cells/ml and then 5 weeks storage at 4 degrees C. The isolate P39 showed to be the most effective for controlling of this disease and decreased the damage by 77.28% respectively. PMID- 18396832 TI - Detection of phlD gene in some fluorescent pseudomonads isolated from Iran and its relative with antifungal activities. AB - Fluorescent pseudomonads that produce antibiotic 2,4-diacetylphloroglocinol (2,4 DAPG) are important group of PGRP that inhibit a broad spectrum of plant pathogenic fungi. Studying on genetic diversity of 2,4-diacetylphloroglucinol producing fluorescent pseudomonads has been shown with special importance. The first step to investigate the genetic diversity of these bacteria is detecting of the genes required for the biosynthesis of this antibiotic. The objectives of the current study were detection of phlD gene within fluorescent pseudomonads by a PCR-based assay, and comparison of phenotypic and genotypic characteristics of fluorescent pseudomonads with proven biocontrol potential against some soil-borne phytopathogenic fungi. We used a collection of 47 fluorescent Pseudomonas spp. some with known biological control activity against Macrophomina phaseolina, Rhizoctonia solani, Phytophthora nicotianae var. parasitica, Pythium sp. and Fusarium sp. in vitro and the potential to produce known secondary metabolites such as, siderophore, HCN and protease. The results indicated that 66, 40.42, 63.82,48.94 and 27.65% of strains revealed antagonistic activity against R. solani, M. phaseolina, Pythium sp., P. nicotianae and Fusarium sp., respectively. Rhizoctonia solani recognized as the most vulnerable fungus. Among 47 strains, 76.59, 97.87 and 17% of strains produced protease, siderophore and HCN, respectively. We could detect phlD gene in strains P-5, P-32, P-47. Strain CHA0 was used as positive control for the detection this gene. Overall, there was no obvious link between the existence of phlD gene and inhibition of fungal growth or production of the antifungal metabolites in vitro. But in some strains such as CHA0 and P-5, we saw a link between the existence of phlD and antifungal activities. Studying on detection and diversity of phlD provides a fundamental knowledge for developing a rapid genetic screening system to identify a potential biocontrol strains. PMID- 18396833 TI - Effect of carbon and nitrogen sources on growth and biological efficacy of Pseudomonas fluorescens and Bacillus subtilis against Rhizoctonia solani, the causal agent of bean damping-off. AB - One of the most important environmental factors that regulate the growth and antagonistic efficacy of biocontrol agents is the medium. The aim of this paper was to find the nitrogen and carbon sources that provide maximum biomass production of strains P-5 and P-6 (Pseudomonas fluorescens), B-3 and B-16 (Bacillus subtilis) and minimum cost of media, whilst maintaining biocontrol efficacy. All of the strains were grown in seven liquid media (pH=6.9) including: sucrose + yeast extract, molasses of sugar beet + yeast extract in 2:1 and 1:1 w/w ratios, molasses of sugar beet + urea, nutrient broth, molasses and malt extract, at an initial inoculation of 1 x 10(5) CFU ml(-1). Cells from over night cultures used to inoculate soil at 1 x 10(9) CFU cm(-3) soil. At the same time, fungal inoculum (infected millet seed with Rhizoctonia solani) was added to soil at the rate of 2 g kg(-1) soil. Results indicated that growth of P-6, B-3 and B 16 in molasses + yeast extract (1:1 w/w) medium was significantly higher than in the other media. Molasses + yeast extract (1:1 and 2:1 w/w) media supported rapid growth and high cell yields in P-5. In greenhouse condition, results indicated that the influence of the media on the biocontrol efficacy of P-5, P-6, B-3 and B 16 was the same and Pseudomonas fluorescens P-5 in molasses and malt extract media reduced the severity of disease up to 72.8 percent. On the other hand, there were observed significant differences on bean growth after one month in greenhouse. P-5 in molasses + yeast extract (1:1 w/w) medium had the most effects on bean growth promotion. In this study molasses media showed good yield efficacy in all of the strains. The high sucrose concentration in molasses justifies the high biomass in all of the strains. Also, the low cost of molasses allows its concentration to be increased in media. On the other hand, yeast extract was the best organic nitrogen source for antagonist bacteria but it is expensive for an industrial process. So it should be replaced by another industrial product instead of yeast extract, which confirm by an economic and technological study. The results obtained in this study could be used to provide a reliable basis to increase the population of biocontrol agents in fermentation process. PMID- 18396834 TI - Effect of timing of application Pseudomonas fluorescens in suppression Sclerotinia sclerotiorum, the causal agent of white mold in canola. AB - Biological control of sclerotinia disease, as an important alternative to chemical control, has received considerable attention due to the lack of resistant varieties in most crop, and increasing concerns over fungicide resistance in population of Sclerotinia sclerotiorum, and fungicide residues in the environment. One biocontrol agent, Pseudomonas fluorescens PB-3, has been showed the antagonistic relationship between itself and S. sclerotiorum was investigated in this study. A petal infection technique was used to detect efficacy of timing of application strain PB-3 in the suppression of S. sclerotiorum on canola. Significant difference in disease severity (p<0.05) were found with respect to timing of ascospore applications in the control treatments (ascospores only). The superior competitive ability strain PB-3 was demonstrated by its complete suppression of disease severity when applied as a co-inoculation treatment or prior to ascospores inoculation. Analysis of effect of applying strain PB-3 after ascospores was indicated that treatment in which strain PB-3 was added to petals 48 or 24 h after ascospores, or when there were no bacteria present at all, had higher rates of disease progression. It would be appear that bacteria are able to significantly inhibit disease when applied before or even at the same time as ascospores. In a practical sense, this could mean that a field application of antagonist could be concurrent with infection by the pathogen. PMID- 18396835 TI - Influence of strobilurin compounds on the development of Puccinia horiana. AB - The effectiveness of 5 strobilurin fungicides: Acanto 250 SC, Amistar 250 SC, Discus 500 WG, Signum 33 WG, Zato 50 WG and Score 250 EC (triazole-standard) in the control of Puccinia horiana was tested on chrysanthemum cv. Melba Blane. Number of pustules per leaf was observed before treatment, as well as 2 and 4 weeks after treatment. Additionally, after 4 weeks, their influence on plant growth, size of pustules and eventually phytotoxicity were assessed. Plants were sprayed preventively and curatively 4 times at weekly intervals. In preventive programme after 4 weeks of investigations, average of about 1.5 spot per leaf was recorded on control plants. Disease symptoms did not appear on chrysanthemums protected by fungicides Signum 33 WG at concentration 0.18% and Zato 50 WG at concentration 0.015%. In case of the other tested fungicides only sporadically pustules on single leaves were noticed (over 95% of them were browned, dried and crumbled). On control plants more than 46% of leaves was infected but in case of the other investigated fungicides it was only 3-7%. Diameter of spots on plants protected with: Score 250 EC, Acanto 250 SC and Discus 500 WG was similar as on the leaves of control plants. On plants protected with Amistar 250 SC the diameter of pustules was significantly lower than on control plants. In curative programme after 4 weeks of tests, leaves of control plants had about 4 spots per leaf. On leaves sprayed with Score 250 EC at concentration 0.05% there were 4 times less spots than on control plants. Chrysanthemums treated with tested strobilurin fungicides had about 2 times less pustules than on control plants. Diameter of spots on control plant leaves was 3.6 mm and was similar as on chrysanthemums protected with tested fungicides. On leaves of control plants all pustules were active without visible changes or damage but in case of the investigated fungicides almost 100% of them were browned, dried and crumbled. After 4 weeks of experiment, a measured height of control plants was about 125 mm and was similar to the height of plants protected with tested fungicides except Signum 33 WG, which stimulated plant growth. None of tested compounds showed phytotoxic effect. PMID- 18396836 TI - The effect of Pythium oligandrum and chitosan used in control of potato against late blight and the occurrence of fungal diseases on tuber peel. AB - The aim of two year investigation was the valuation the effect of biopreparate Polyversum (B.A.S. Pythium oligandrum) and preparate Biochikol 020 PC (B.A.S. chitosan) used in control of potato against Phytophthora blight on the tuber infestation during storage by Helminthosporium solani and tuber infestation by sclerotia of Rhizoctonia solani. As the standard fungicide Vitavax 200 FS (B.A.S. karboxin and thiuram) was used. After harvesting 100 tubers from each plots was collected and put in storage. The analysis of tuber infestation by Rhizoctonia solani and Helminthosporium solani was made after harvesting (September) and later every 3 months during storage period (December, March). The percent of diseased tubers in tested sample and also infestation degree of bulbs using 5 degree scale was estimated. The received results of investigations ascertained, that all tested preparations during potato vegetation influenced on lower (in comparison with control) degree of bulbs infestation by sclerots of Rhizoctonia solani and the mean degree of infestation by Helminthosporium solani. Moreover the percent of diseased tubers infected by pathogens with tested preparations combination was significant lower than in control. PMID- 18396837 TI - Isolation and evaluation of bacteria and fungi as biological control agents against Rhizoctonia solani. AB - Rhizoctonia solani is one of the most important limiting factors for potato production and storage in Belgium and worldwide. Its management is still strongly dependent on chemical treatments. The aim of this work was to evaluate the possibility of exploiting bacteria and fungi in order to control this pathogen. Among a collection of 220 bacterial strains isolated from different organs of healthy potato plants and rhizospheric soils, 25 isolates were selected using screening methods based on in vitro dual culture assays. The mycelial growth inhibition rate of the pathogen was ranged from 59.4 to 95.0%. Also seven fungal strains isolated from the rhizospheric soil and potato roots showed a highly mycelial growth inhibition of R. solani. The mycelial growth inhibition rate obtained with these fungi was included between 60.0 and 99.4%. From this preliminary study, the further investigations will be planned to determine the bacterial isolates systematic, species of fungal strains by using molecular tools and to assess their efficacy against R. solani in greenhouse trials. PMID- 18396838 TI - Induction of systemic defense responses in rice against the sheath blight pathogen Rhizoctonia solani, by means of riboflavin. PMID- 18396839 TI - Study of the aggressiveness of Rhizoctonia solani isolates. AB - This paper presents an in vitro test to screen the pathogenicity of different Rhizoctonia solani isolates on a host range. The level of aggressivity of the different isolates was different for several host plants tested. There were significant differences between the crops and the isolates tested. In general, the disease level was higher on beans, lettuce and cabbage. In carrot and rye grass the level of infection was lower for the isolates of R. solani tested. The potato isolates of R. solani were less aggressive than the isolates coming from maize, fodder beet and sugar beet. The R. solani isolates were also biochemically characterized by pectic zymograms: the isolates Rs0401 (from maize) and Rs0504 (from sugar beet) belong both to the anastomosis group AG2-2. PMID- 18396840 TI - Biological control of Sclerotinia sclerotiorum (Lib.) de Bary, the causal agent of white mold, by Pseudomonas species on canola petals. AB - Sclerotinia sclerotiorum is an important pathogen on canola. Due to the public concern over pesticide use, alternative methods of disease control, such as biological control, should be considered. Several bacterial strains were isolated from canola and soja plants. Inhibition of S. sclerotiorum by bacterial strains in vitro was assayed on PDA medium in dual culture test. Eight Pseudomonas sp. strains (PB-3, PB-4, PB-5, PB-6, PB-7, PB-8, PB-10 and PB-11) caused inhibition zone against 5. sclerotiorum hyphal growth. The biocontrol potential of the bacteria was tested in a plant assay. Disease suppression was investigated using a petal inoculation technique. Canola petals were pretreated with bacteria, and then inoculated with 5. sclerotiorum ascospores 24 h later. Greenhouse experiment showed that application of Pseudomonas sp. strains (1 x 10(8) cfu ml(-1)) effectively suppressed S. sclerotiorum (1 x 10(5) ascospores ml(-1)) on petals and all of them achieved significant (P<0.01) disease suppression. Fourteen days after inoculation, strain PB-3 had 88/7% disease control and strain PB-4 had 69/9% disease control. Result from all studies indicates PB-3 to be effective biocontrol against S. sclerotiorum of canola. PB-3, PB-4, PB-7, PB-8, PB-10 and PB-11 were identified as Pseudomonas fluorescens biovar III. PB-5 and PB-6 was identified as Pseudomonas fluorescens biovar II. Strains PB-3, PB-4, PB-6, PB-10 and PB-11 produced protease and HCN. Strain PB-5 produce protease; no HCN. PMID- 18396841 TI - Study on the sensitivity of Venturia inaequalis to anilinopyrimidine fungicides in Italy. PMID- 18396842 TI - Management of apple scab (Venturia inaequalis) in Romania based on electronic warnings. AB - Apple scab, Venturia inaequalis (Cke) Wint., is a major problem in Romanian apple orchards and is responsible for serious economic losses. The purchase of spraying equipment and phytopharmaceutical products is very expensive in comparison to the income of local apple growers. This is one of the reasons why reduced fungicide sprayings are becoming more and more important. That's why a project was started to investigate the influence of West-European control measures and to improve interaction between local apple growers and research facilities. This experiment took place in an orchard of the university of Timisoara (USABT), where occurrence of apple scab was very high, and in a well managed commercial orchard, both in the Banat region. In July 2006, a comparative study between a traditional Romanian management program, a calendar-based program and a computerized weather based management program was conducted. Four test plots were taken. The sprayments in plot 1 happened according to the integrated computerized model. No sprayings took place in plot 2. In plot 3 were some sprayments done with sulphur according to the traditional Romanian program. The sprayings in plot 4 were done following a calendar-based spraying program. The products that were used for the sprayments were all easy to obtain in Romania such as dodine (Sillit flow), dithianon (Delan flow), difenoconazole (Score), pyrimethanil (Scala), metiram (Polyram), kresoxim-methyl (Stroby). The occurrence of scab in plot 1 was very low in comparison to the other plots. Analysing the data, on average, the treatments done in plot 1 caused about 50% less damage than the traditional sprayings. This indicated that with a relative small amount of well timed sprayings a enormous amount of damage can be prevented. A calendar-based management program seems to be less effective (more sprayings needed and the results are worse). These results also emphasize the importance of the development of an adequate apple scab management program that should be presented to local apple growers. PMID- 18396843 TI - Compatibility of Verticillium lecani (Zimm.) with several pesticides. AB - The entomopathogenic fungus, Verticillium lecanii (Zimm.) Viegas is a widespread hyphomycete consisting of a heterogeneous species aggregate. Conidia survival may be effected by environmental factors or by bio-pesticides and chemical products used to protect crop plants. In this research compatibility of mentioned fungi with imidachlopride, flufenoxuron, teflubenzuron+phozalone, endosulfan, amitraz, and effect of these pesticides on vegetative growth of the fungus were studied. The formulations of pesticides were tested in three concentration (mean concentration-MC, half MC and twice the MC) and using Duncan multiple range test, treatment were grouped. Effects of the pesticides on V. lecanii vegetative growth in concerned results have shown that almost all formulations studies significantly inhibition fungal development. The results indicated that flufenoxuron (at three concentrations), teflubenzuron+phozalone (at MC and twice the MC), induced levels of vegetative growth inhibition higher than 68%. Only data on amitraz and imidachlopride (at half MC) were not significantly different from the control, which is indicated the compatible formulations with V. lecanii. These formulations could be used simultaneously with this entomopathogenic in integrated pest management. PMID- 18396844 TI - Study of interaction between plant nutrition (N,P,K and Ca) and verticillios wilt disease (Verticillium dahliae) in cucumber. AB - Cucumber wilt disease caused by verticillium dahliae is one of the most important diseases of cucumber. Disease severity can to be decreased by plant nutrition management, especially nitrogen, phosphorus, potassium and calcium levels. For this purpose, interaction between plant nutrition and cucumber verticillios disease in cucumber was studied. Different levels of N, P, K and Ca on one current cultivar of cucumber were investigated at the presence and absence of fungus, using a factorial analysis based on completely randomized design with three replications. Significant different were observed among the different levels of elements used in this study. Presence of N, P, K and Ca increased the percentage of alive plants in comparison with control (distilled water + fungus). The effects of different levels of N, P, K and Ca were also investigated on growth of V. dahliae in vitro at a complete randomized design with three replications. Variance analysis of growth area of the fungus showed significant difference among some levels of the elements. The highest area growth of fungus was occurred at 0.1 gr/l of KNO3 and was 47/26 Cm2. The factors investigated, clearly suggested the occurrence of a wide interaction between cucumber nutrition (N, P, K and Ca) and V.dahliae and showed the importance of plant nutrituin management for decrease of disease severity. PMID- 18396845 TI - Three years survey of Tomato yellow leaf curl Sardinia virus reservoir weed hosts in southern Italy. AB - During the period from August 2004 to June 2006 a serious tomato yellow leaf curl epidemic caused by both Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCS) was observed in protected tomato crops in Castrovillari, Calabria Region, in a group of greenhouses where tomato is grown hydroponically. A three years survey for reservoir weed hosts of these viruses was performed during summer period in order to identify where the viruses persist during the host-free period, interesting an area covering a ray of 500 m around the group of greenhouses. About 350 samples were collected from symptomless and symptomatic plants of the following botanic families: Graminaceae, Compositeae, Solanaceae, Portulacaceae, Malvaceae, Chenopodiaceae, Amaranthaceae, Convolvulaceae, Brassicaceae, Labiatae, Plantaginaceae, Asteraceae. Any virus presence was evaluated by DAS ELISA, using a "broad-spectrum" reagent combination detecting different Begomoviruses including TYLCSV and TYLCV. A couple of synthetic oligonucleotides allowing the amplification of the whole coat protein (CP) gene was used for PCR of ELISA positive samples in order to perform the molecular characterisation of the viral isolate responsible of the disease. RFLP analysis performed on the PCR product, 1008 bp long, showed the presence of only TYLCSV in the weeds found infected and belonging to Sonchus asper, Solanum nigrum, Datura stramonium and Cardaria draba species. Similarity analysis performed between the CP of each isolate and the TYLCSV isolate recovered within the greenhouse and responsible of the epidemic in mixed infection with a TYLCV isolate resulted in a value of 100% of identity, thus indicating that there was no variability in TYLCSV population in the surveyed area. S. asper, S. nigrum, D. stramonium and C. draba, as alternative hosts of TYLCSV and nutrient plants of the virus vector, Bemisia tabaci, were found to play an important role in virus ecology and epidemiology in the studied tomato ecosystem. No weed between those investigated has been found to be infected by TYLCV so far. To our knowledge this is the first report of S. asper and C. draba as TYLCSV hosts in natural infection. PMID- 18396846 TI - Relationship between Tomato yellow leaf curl viruses and the whitefly vector. AB - Tomato Yellow Leaf Curl Virus (TYLCV) and Tomato Leaf Curl Virus (ToLCV) are the currently known begomoviruses in Uganda. The relationship with their whiteflies (Bemisia tabaci) vector and its management were not known in Uganda. A direct relationship was expected between these begomoviruses and whiteflies at Buwama in Mpigi district of Uganda. Farmer practices were expected to have limited efficacy. To investigate this, a completely randomised block design was used for all trials with six treatments replicated three times. Treatments evaluated were farmer whitefly pest management practices. These included both chemical pesticide and non-pesticide applications. Data on whitefly population and tomato yellow leaf curl virus disease incidence was recorded weekly, and analysed using SAS and SPSS statistical programmes for ANOVA, and correlations. Ranked means, coefficients of variation and standard errors were noted. Virus-vector relationship field studies established that virus occurrence varied in space and time, and with management practices, crop development stage, and weather conditions. A negative relationship (R = -0.14, p 0.04) was established between number of plants infected with TYLCV (sensu lato) and percentage marketable tomato yield. Tomato maturity was inversely propotional to whitefly infestation (R = -0.5, p 0.0001). Uprooting and application of dimethoate was most effective of the six treatments. Tomato leaf curl virus diseases and whitefly management options were established in Uganda. PMID- 18396847 TI - [How many children and adolescents do receive a combined behavioral and psychopharmacological therapy?]. AB - So far in German literature no empirical data are present concerning parallel medications of behavioral therapy. Also the international research does not treat this topic sufficiently. For this reason this work examines on the basis from reports of requests for behavioral therapy, which medications or groups of substances were used for which child and adolescent psychiatric disorders treated by behavioral therapy. It examines additionally whether correlations of such combinations exist to specific socio-demographic or school-characteristics. Out of 1021 first and subsequent requests for behavioral therapy from the years 2001 variables were retrospectively evaluated regarding intear alia diagnosis, sociodemographic data, school situation, somatic comorbidities or therapies and therapy indication. Results reveal out of five children or adolescents one has an behavioral therapy plus an additional psychopharmacological treatment. 80% of these medications are psychostimulants. Half of the children and adolescents suffering from ADHD or conduct disorders are treated with such, while diagnosis of other domains contribute to combined therapies only with a small percentage. Differences between non-treated and psychopharmacologically treated children and adolescents with ADHD or conduct disorders are presented. PMID- 18396848 TI - [Promotion of resilience in institutions of early childhood education (kindergarten)--first results of a prevention project]. AB - This article focuses on first results of a prevention project dealing with the promotion of resilience in institutions of early childhood education (e.g. kindergarten). In four institutions with 278 children the early childhood educators are trained and the children themselves take part in a special training programme. Additionally the parents are given the chance to participate in special courses and individual counselling, while the institutions are involved in networks with other institutions, e.g. for educational guidance. The process and the effects of the programme are evaluated in a control group design by qualitative and quantitative (standardised tests) methods. After a first project stage of six months interim findings show a positive response in all target groups. There are significant positive results on the children of the treatment group in their self-concept and in their cognitive development. PMID- 18396849 TI - [The handling of custodial criteria by expert evaluators and the expectations for solution-oriented evaluations from the perspective of Austrian family court judges]. AB - There has been increasing discussion of a paradigm shift in expert evaluations of family psychology during custodial proceedings for a long time, suggesting that status-diagnostic evaluation should give way to working together with the parents with a "solution-oriented" process approach. An Austrian-wide evaluation of the quality of current evaluation practice indicated that a majority of the experts, and especially the judges in family courts, endorsed this "solution-oriented" approach for expert evaluators; the judges primarily in the expectation that this will lead both to a greater acceptance and sustainability of their decisions on custodial and visiting rights and that it could also, over the long-term, serve to increase the efficiency of the legal process. However, if the evaluation modalities are altered accordingly, proven custodial criteria should still remain in the foreground. In addition to the child's attachment, family judges and experts, working in unison, assign continuity and the parents' own parenting abilities the next greatest importance within the framework of individual custodial criteria. Psychometric diagnostic procedures should still be used when establishing a solution-oriented expert evaluation procedure in order to comprehensively map out the needs of the child as well as individual aspects of the parents' own personality in particular, where a joint solution cannot be achieved. PMID- 18396850 TI - [Effectiveness of in-home family-focused interventions]. AB - Results of evaluation studies with a (quasi-)experimental design covering different models of in-home family-focused interventions are reviewed. These models are of the family support or the family preservation type of program. The methodological quality of the evaluation is high or very high in the case of the two family support programs Nurse Home Visitation and Healthy Families America and the two family preservation programs Functional Family therapy and Multisystemic Therapy. All of them are producing clinically relevant positive long-term effects. The models of in-home family-focused intervention currently used in Germany are to be evaluated in studies with (quasi-)experimental design. PMID- 18396851 TI - Calorimetric study of correlated disordering in [Hdamel]2[Cu(II)(tdpd)2] x 2 THF crystal. AB - The heat capacity of [Hdamel]2[Cu(II)(tdpd)2] x 2 THF was measured from 6 to 250 K by adiabatic calorimetry. There are four heat anomalies around 150 K associated with disordering in the orientation of the uncoordinated THF molecules and in the conformation of the out-of-plane allyl groups of [Hdamel](+) units. The total entropy of transition was determined to be 19.8 J K(-1) mol(-1), less than the 4R ln 2 (R = gas constant) expected from the crystal structure at room temperature. The smallness of the total entropy change on phase transitions proves the presence of the strong motional correlation between the adjacent allyl groups. The calorimetric conclusion agreed with the crystal structure at 200 K re examined in this study. PMID- 18396852 TI - Addition reactions of alkyl and carboxyl radicals to vinylidene fluoride. AB - The addition reactions of alkyl radicals CF3* and CH3* and carboxyl radicals C2H5O*, C2H5OCOO*, CF3COO*, and CH3COO* to a vinylidene fluoride (VDF) molecule are studied using ab initio calculations. These radicals were selected because they are intermediate or final products of diacyl peroxides decomposition in the initiation reactions of VDF polymerization. Two combinations of methods for energetics and structure optimization are applied: QCISD/6-311G(d,p)//HF/6-31G(d) and B3LYP/6-311G+(3df, 2p)//B3LYP/6-31G(d). It is found that the formed bond length of the product, the forming bond length of the transition state, and the attack angle of the product structures are not sensitive to the level of theory even though the attack angle of the transition state structures is. Early transition states are obtained upon attack at both high-substituted and nonsubstituted carbon atom VDF ends. Kinetic and thermodynamic control rules play different roles on governing the reactivity of the addition with the studied radicals. Both theoretical methods yield the same trends for the preferential attack site in terms of regioselectivity, barrier energies, and reaction enthalpies. It is shown that the addition reactions of the intermediate radicals C2H5OCOO*, CF3COO*, and CH3COO* of the decomposition of diethyl peroxydicarbonate, trifluoroacetyl peroxide, and diacetyl peroxide initiators yield smaller energy barriers than the additions of the corresponding final radicals, C2H5O*, CF3*, and CH3*; therefore, the reactions of the intermediate radicals should not be ignored when analyzing the initiation process of the VDF polymerization using those initiators. PMID- 18396853 TI - Comparative DFT study of crystalline ammonium perchlorate and ammonium dinitramide. AB - The electronic structure, vibrational properties, absorption spectra, and thermodynamic properties of crystalline ammonium perchlorate (AP) and ammonium dinitramide (ADN) have been comparatively studied using density functional theory in the local density approximation. The results shows that the p states for the two solids play a very important role in their chemical reaction. From the low frequency to high frequency region, ADN has more motion modes for the vibrational frequencies than AP. The absorption spectra of AP and ADN display a few, strong bands in the fundamental absorption region. The thermodynamic properties show that ADN is easier to decompose than AP as the temperature increases. PMID- 18396854 TI - Molecular characteristics for solid-state limited solubility. AB - Solubility and solid-state characteristics were determined and multivariate data analysis was used to deduce structural features important for solid-state limited solubility of marketed drugs. Molecules with extended ring structures and large conjugated systems were less soluble, indicating that structural features related to rigidity and aromaticity result in solubility restricted by stable crystal structures. These descriptors successfully predicted the applied test set and can be useful for avoiding synthesis of compounds behaving like "brick dust". PMID- 18396855 TI - Synthesis and structure-activity relationships of 4-pyridones as potential antimalarials. AB - A series of diaryl ether substituted 4-pyridones have been identified as having potent antimalarial activity superior to that of chloroquine against Plasmodium falciparum in vitro and murine Plasmodium yoelii in vivo. These were derived from the anticoccidial drug clopidol through a systematic study of the effects of varying the side chain on activity. Relative to clopidol the most active compounds show >500-fold improvement in IC50 for inhibition of P. falciparum in vitro and about 100-fold improvement with respect to ED50 against P. yoelii in mice. These compounds have been shown elsewhere to act selectively by inhibition of mitochondrial electron transport at the cytochrome bc1 complex. PMID- 18396857 TI - Synthesis and biological activity of fluorinated combretastatin analogues. AB - With the aim of understanding the influence of fluorine on the double bond of the cis-stilbene moiety of combretastatin derivatives and encouraged by a preliminary molecular modeling study showing a different biological environment on the interaction site with tubulin, we prepared, through various synthetic approaches, a small library of compounds in which one or both of the olefinic hydrogens were replaced with fluorine. X-ray analysis on the difluoro-CA-4 analogue demonstrated that the spatial arrangement of the molecule was not modified, compared to its nonfluorinated counterpart. SAR analysis confirmed the importance of the cis stereochemistry of the stilbene scaffold. Nevertheless, some unpredicted results were observed on a few trans-fluorinated derivatives. The position of a fluorine atom on the double bond may affect the inhibition of tubulin polymerization and cytotoxic activity of these compounds. PMID- 18396858 TI - Novel approach to structure-based pharmacophore search using computational geometry and shape matching techniques. AB - Computationally efficient structure-based virtual screening methods have recently been reported that seek to find effective means to utilize experimental structure information without employing detailed molecular docking calculations. These tools can be coupled with efficient experimental screening technologies to improve the probability of identifying hits and leads for drug discovery research. Commercial software ROCS (rapid overlay of chemical structures) from Open Eye Scientific is such an example, which is a shape-based virtual screening method using the 3D structure of a ligand, typically from a bound X-ray costructure, as the query. We report here the development of a new structure based pharmacophore search method (called Shape4) for virtual screening. This method adopts a variant of the ROCS shape technology and expands its use to work with an empty crystal structure. It employs a rigorous computational geometry method and a deterministic geometric casting algorithm to derive the negative image (i.e., pseudoligand) of a target binding site. Once the negative image (or pseudoligand) is generated, an efficient shape comparison algorithm in the commercial OE SHAPE Toolkit is adopted to compare and match small organic molecules with the shape of the pseudoligand. We report the detailed computational protocol and its computational validation using known biologically active compounds extracted from the WOMBAT database. Models derived for five selected targets were used to perform the virtual screening experiments to obtain the enrichment data for various virtual screening methods. It was found that our approach afforded similar or better enrichment ratios than other related methods, often with better diversity among the top ranking computational hits. PMID- 18396859 TI - In situ formation of heterobimetallic salen complexes containing titanium and/or vanadium ions. AB - A combination of high-resolution electrospray mass spectrometry and (1)H NMR spectroscopy has been used to prove that when a mixture of [(salen)TiO]2 complexes containing two different salen ligands (salen and salen') is formed, an equilibrium is established between the homodimers and the heterodimer [(salen)TiO2Ti(salen')]. Depending upon the structure and stereochemistry of the two salen ligands, the equilibrium may favor either the homodimers or the heterodimer. Extension of this process to mixtures of titanium(salen) complexes [(salen)TiO]2 and vanadium (V)(salen') complexes [(salen')VO] (+)Cl (-) allowed the in situ formation of the heterobimetallic complex [(salen)TiO2V(salen')] (+)X (-) to be confirmed for all combinations of salen ligands studied except when the salen ligand attached to titanium contained highly electron-withdrawing nitro groups. The rate of equilibration between heterobimetallic complexes is faster than that between two titanium complexes as determined by line broadening in the (1)H NMR spectra. These structural results explain the strong rate-inhibiting effect of vanadium (V)(salen) complexes in asymmetric cyanohydrin synthesis catalyzed by [(salen)TiO]2 complexes. It has also been demonstrated for the first time that the titanium and vanadium complexes can undergo exchange of salen ligands and that this is catalyzed by protic solvents. However, the ligand exchange is relatively slow (occurring on a time scale of days at room temperature) and so does not complicate studies aimed at using heterobimetallic titanium and vanadium salen complexes as asymmetric catalysts. Attempts to obtain a crystal structure of a heterobimetallic salen complex led instead to the isolation of a trinuclear titanium(salen) complex, the formation of which is also consistent with the catalytic results obtained previously. PMID- 18396860 TI - 1,2,4,5-benzenetetracarboxylate- and 2,2'-bipyrimidine-containing cobalt(II) coordination polymers: preparation, crystal structure, and magnetic properties. AB - Three new mixed-ligand cobalt(II) complexes of formula [Co2(H2O)6(bta)(bpym)]n.4nH2O (1), [Co2(H2O)2(bta)(bpym)]n (2), and [Co2(H2O)4(bta)(bpym)]n.2nH2O ( 3) (bpym = 2,2'-bipyrimidine and H 4bta = 1,2,4,5 benzenetretracaboxylic acid) have been synthesized and characterized by single crystal X-ray diffraction. 1 is a chain compound of mer-triaquacobalt(II) units which are linked through regular alternating bis-bidentate bpym and bis monodentate bta groups. 2 and 3 are three-dimensional compounds where aquacobalt(II) ( 2) and cis-diaquacobalt(II) ( 3) entities are linked by bis bidentate bpym ( 2 and 3) and tetrakis- ( 2 and 3) and octakis-monodentate ( 2) bta ligands. The cobalt atoms in 1- 3 exhibit somewhat distorted octahedral surroundings. Two bpym-nitrogen atoms ( 1- 3) and either two bta-oxygens ( 2) or one bta-oxygen and a water molecule ( 1 and 3) build the equatorial plane, whereas the axial positions are filled either by two water molecules ( 1) or by a bta-oxygen atom and a water molecule ( 2 and 3). The values of the cobalt-cobalt separation across the bridging bpym vary in the range 5.684(2)-5.7752(7) A, whereas those through the bta bridge cover the ranges 5.288(2)-5.7503(5) A (across the anti-syn carboxylate) and 7.715(3)-11.387(1) A (across the phenyl ring). The magnetic properties of 1- 3 have been investigated in the temperature range 1.9-290 K. They are all typical of an overall antiferromagnetic coupling with the maxima of the magnetic susceptibility at 14.5 ( 1) and 11.5 K ( 2 and 3). Although exchange pathways through bis-bidentate bpym and carboxylate-bta in different coordination modes are involved in 1- 3, their magnetic behavior is practically governed by that across the bpym bridge, the magnitude of the exchange coupling being J = -5.59(2) ( 1), -4.41(2) ( 2), and -4.49(2) ( 3) with the Hamiltonian H = - JS 1 S 2. PMID- 18396861 TI - In search of the [PhB(mu-N(t)Bu)2]2As. radical: experimental and computational investigations of the redox chemistry of group 15 bis-boraamidinates. AB - DFT calculations for the group 15 radicals [PhB(mu-N(t)Bu)2]2M. (M = P, As, Sb, Bi) predict a pnictogen-centered SOMO with smaller contributions to the unpaired spin density arising from the nitrogen and boron atoms. The reactions of Li 2[PhB(mu-NR)2] (R = (t)Bu, Dipp) with PCl 3 afforded the unsolvated complex LiP[PhB(mu-N(t)Bu)2] 2 ( 1a) in low yield and ClP[PhB(mu-NDipp)2] (2), both of which were structurally characterized. Efforts to produce the arsenic-centered neutral radical, [PhB(mu-N (t) Bu) 2] 2As., via oxidation of LiAs[PhB(mu N(t)Bu)2]2 with one-half equivalent of SO 2Cl 2, yielded the Zwitterionic compound [PhB(mu-N (t) Bu) 2As(mu-N(t)Bu)2B(Cl)Ph] (3) containing one four coordinate boron center with a B-Cl bond. The reaction of 3 with GaCl3 produced the ion-separated salt, [PhB(mu-N(t)Bu)2] 2As (+)GaCl 4 (-) ( 4), which was characterized by X-ray crystallography. The reduction of 3 with sodium naphthalenide occurred by a two-electron process to give the corresponding anion [{PhB(mu-N(t)Bu)2} 2As] (-) as the sodium salt. Voltammetric investigations of 4 and LiAs[PhB(mu-N (t) Bu) 2] 2 ( 1b) revealed irreversible processes. Attempts to generate the neutral radical [PhB(mu-N(t)Bu)2] 2As. from these ionic complexes via in situ electrolysis did not produce an EPR-active species. PMID- 18396862 TI - Copper dioxygen adducts: formation of bis(mu-oxo)dicopper(III) versus (mu 1,2)Peroxodicopper(II) complexes with small changes in one pyridyl-ligand substituent. AB - The preference for the formation of a particular Cu 2O 2 isomer coming from (ligand)-Cu (I)/O 2 reactivity can be regulated with the steric demands of a TMPA (tris(2-pyridylmethyl)amine) derived ligand possessing 6-pyridyl substituents on one of the three donor groups of the tripodal tetradentate ligand. When this substituent is an -XHR group (X = N or C) the traditional Cu (I)/O 2 adduct forms a (mu-1,2)peroxodicopper(II) species ( A). However, when the substituent is the slightly bulkier XR 2 moiety {aryl or NR 2 (R not equal H)}, a bis(mu oxo)dicopper(III) structure ( C) is favored. The reactivity of one of the bis(mu oxo)dicopper(III) species, [{(6tbp)Cu (III)} 2(O (2-)) 2] (2+) ( 7-O 2 ) (6tbp = (6- (t)Bu-phenyl-2-pyridylmethyl)bis(2-pyridylmethyl)amine), was probed, and for the first time, exogenous toluene or ethylbenzene hydrocarbon oxygenation reactions were observed. Typical monooxygenase chemistry occurred: the benzaldehyde product includes an 18-O atom for toluene/ 7- (1) (8)O 2 reactivity, and a H-atom abstraction by 7-O 2 is apparent from study of its reactions with ArOH substrates, as well as the determination of k H/ k D approximately 7 in the toluene oxygenation (i.e., PhCH 3 vs PhCD 3 substrates). Proposed courses of reaction are presented, including the possible involvement of PhCH 2OO (*) and its subsequent reaction with copper(I) complex, the latter derived from dynamic solution behavior of 7-O 2 . External TMPA ligand exchange for copper in 7-O 2 and O-O bond (re)formation chemistry, along with the ability to protonate 7-O 2 and release of H 2O 2 indicate the presence of an equilibrium between [{(6tbp)Cu (III)} 2(O (2-)) 2] (2+) ( 7-O 2 ) and a (mu-1,2)peroxodicopper(II) form. PMID- 18396863 TI - Synthesis, Raman spectra and crystal structures of [Cu(XeF2)n](SbF6)2 (n=2, 4). AB - Pure [Cu(XeF2)2](SbF6)2 was prepared by the reaction of Cu(SbF 6) 2 with a stoichiometric amount of XeF2 in anhydrous hydrogen fluoride (aHF) at ambient temperature. The reaction between Cu(SbF6)2 and XeF2 (1:4 molar ratio) in aHF yielded [Cu(XeF2)4](SbF6)2 contaminated with traces of Xe 2F 3SbF6 and CuF2. The 6-fold coordination of Cu(2+) in [Cu(XeF2)2](SbF6)2 includes two fluorine atoms from two XeF2 ligands and four fluorine atoms provided by four [SbF6](-) anions. The neighboring [Cu(XeF 2)2](2+) moieties are connected via two [SbF6] units, with the bridging fluorine atoms in cis positions, into infinite [Cu(eta(1) XeF2)2](cis-eta(2)-SbF 6)2[Cu(eta(1)-XeF 2)2] chains. Because of the high electron affinity of Cu(2+), coordinated XeF2 shows the highest distortion (Xe Fb=210.2(5) pm, Xe-Ft=190.6(5) pm) observed so far among all known [M(x+)(XeF2)n](A)x (A=BF4, PF6, etc.) complexes. The four equatorial coordination sites of the Cu(2+) ion in [Cu(XeF 2) 4](SbF6)2 are occupied by four XeF 2 ligands. Two fluorine atoms belonging to two [SbF6] units complete the Cu (2+) coordination environment. The neighboring [Cu(XeF2)4](2+) species are linked via one [SbF6] unit, with bridging fluorine atoms in trans positions, into linear infinite [Cu(eta(1)-XeF2)4](trans-eta(2)-SbF6)[Cu(eta(1)-XeF2)4] chains. To compensate for the remaining positive charge, crystallographically independent [SbF6](-) anions are located between the chains and are fixed in the crystal space by weak Xe...F(Sb) interactions. PMID- 18396864 TI - Ruthenium complexes with tetraphenylimidodiphosphinate: syntheses, structures, and applications to catalytic organic oxidation. AB - Treatment of Ru(PPh3)3Cl2 with K(tpip) (tpip(-)=[N(Ph2PO)2](-)) afforded Ru(tpip)(PPh3)2Cl (1), which reacted with 4- t-Bu-C6H4CN, SO2(g), and NH 3(g) to give Ru(tpip)(PPh3)2Cl(4- t-BuC6H4CN) (2), Ru(tpip)(PPh3)2Cl(SO2) (3), and fac [Ru(NH3)3(PPh3)2Cl][tpip] (4), respectively. Reaction of [Ru(CO)2Cl2] x with K(tpip) in refluxing tetrahydrofuran (THF) led to isolation of the K/Ru bimetallic compound K 2Ru2(tpip)4(CO)4Cl2 (5). Photolysis of cis-Ru(tpip) 2(NO)Cl in MeCN and wet CH 2Cl 2 afforded cis-Ru(tpip) 2(MeCN)Cl ( 6) and cis Ru(tpip)2(H2O)Cl (7), respectively. Refluxing 6 in neat THF yielded Ru(tpip) 2(THF)Cl (8). Treatment of Ru(CHR)Cl2(PCy3)2 (Cy=cyclohexyl) with [Ag(tpip)] 4 afforded cis-Ru(tpip)2(CHR)(PCy3) [R=Ph (9), OEt (10)]. Complex 9 is capable of catalyzing oxidation of alcohols and olefins with N-methylmorpholine N-oxide and iodosylbenzene, respectively. The crystal structures of 2-7 and 9 were determined. PMID- 18396865 TI - Tetranuclear [{Ni(HL3)}{W(CN)8}]2 square: a case of antiferromagnetic {NiIIWV} interactions. AB - A tetranuclear cyano-bridged [{Ni(HL3)}{W(CN)8}]2 compound in a square geometry was formed by self-assembling of {W(CN)8}3- and {NiL3}2+ (L3=pentadentate ligand). The structure of the compound has been established by single crystal X ray diffraction. The coordination sphere of the Ni ions is severely distorted with the macrocyclic ligand adopting a facial coordination with only four linkages to the metal center. The N atom of the pendant aminopropyl arm of L3 is no longer coordinated to the metal center but has undergone protonation during the assembling process. Magnetic measurements have revealed an unexpected antiferromagnetic behavior (J=-9 cm(-1)), which has been explained using a microscopic many-body electronic model Hamiltonian, based on DFT results. The many-body model is used to fit both the chiMT versus T and the M versus H plots obtained from experiments. PMID- 18396867 TI - Homoleptic mononuclear and binuclear osmium carbonyls Os(CO)n(n = 3-5) and Os2(CO)n (n = 8, 9): comparison with the iron analogues. AB - The structures and energetics of the experimentally known Os(CO)n ( n = 3-5), Os2(CO)9, and Os2(CO)8 have been investigated using density functional theory. For Os(CO)5, the lowest-energy structure is the singlet D(3h) trigonal bipyramid. However, the C(4v) square pyramid for Os(CO)5 lies only approximately 1.5 kcal/mol higher in energy, suggesting extraordinary fluxionality. For the coordinatively unsaturated Os(CO)4 and Os(CO)3, a D(2d) strongly distorted tetrahedral structure and a Cs bent T-shaped structure are the lowest-energy structures, respectively. For Os2(CO)9, the experimentally observed singly bridged Os2(CO)8(mu-CO) structure is the lowest-energy structure. A triply bridged Os2(CO)6(mu-CO)3 structure analogous to the known Fe2(CO)9 structure is a transition state rather than a true minimum and collapses to the singly bridged global minimum structure upon following the corresponding normal mode. An unbridged (OC)5Os --> Os(CO)4 structure with a formal Os --> Os dative bond analogous to known stable complexes of the type (R3P)2(OC)3Os --> W(CO)5 is also found for Os2(CO)9 within 8 kcal/mol of the global minimum. The global minimum for the coordinatively unsaturated Os2(CO)8 is a singly bridged (OC)4Os(mu CO)Os(CO)3 structure derived from the Os2(CO)9 global minimum by loss of a terminal carbonyl group. However, the unbridged structure for Os2(CO)8 observed in low-temperature matrix experiments lies only approximately 1 kcal/mol above this global minimum. In all cases, the triplet structures for these osmium carbonyls have significantly higher energies than the corresponding singlet structures. PMID- 18396866 TI - Growth mechanism and optical properties of aligned hexagonal ZnO nanoprisms synthesized by noncatalytic thermal evaporation. AB - Vertically aligned perfectly hexagonal-shaped ZnO nanoprisms have been grown on a Si(100) substrate via a noncatalytic thermal evaporation process by using metallic zinc powder in the presence of oxygen gas. The as-grown nanoprisms consist of ultra smooth Zn-terminated (0001) facets bounded with the {0110} surfaces. The as-synthesized products are single-crystalline with the wurtzite hexagonal phase and grown along the [0001] direction, as confirmed from the detailed structural investigations. The presence of a sharp and strong nonpolar optical phonon high-E2 mode at 437 cm(-1) in the Raman scattering spectrum further confirms good crystallinity and wurtzite hexagonal phase for the as-grown products. The as-grown nanoprisms exhibit a strong near-band-edge emission with a very weak deep-level emission in the room-temperature and low-temperature photoluminescence measurements, confirming good optical properties for the deposited products. Moreover, systematic time-dependent experiments were also performed to determine the growth process of the grown vertically aligned nanoprisms. PMID- 18396868 TI - Ni(II)-mediated self-assembly of artificial beta-dipeptides forming a macrocyclic tetranuclear complex with interior spaces for in-line molecular arrangement. AB - Metal-mediated self-assembly of bioinspired molecular building blocks shows promise as an excellent strategy to provide well-defined metal arrays and nanoscopic metallo-architectures in a programmable way. Herein, we report Ni(II) mediated self-assembly of artificial beta-dipeptides (1) which were prepared from a newly designed beta-amino acid bearing a propanediamine ligand as the side chain. The beta-dipeptide (1) has thus two sets of ligands, that is, each building block serves as a tridentate ligand with a bidentate propanediamine unit and an amide carbonyl group. Both C- and N-terminal tridentate ligands in 1 bind to two Ni(II) ions independently, and consequently, four beta-dipeptides are circularly arranged in a head-to-tail fashion to form a macrocyclic tetranuclear Ni(II) complex, Ni414(ClO4)8(H2O)10. The cyclic structure was determined by X-ray analysis and ESI-TOF mass spectrometry. The resulting unique twisted-boat structure allows the formation of isolated spaces for in-line hydrogen-bonded arrangement of water and anion molecules within a hole and two grooves rich in hydrogen bonding groups. PMID- 18396869 TI - Synthesis of Cu-In-S ternary nanocrystals with tunable structure and composition. AB - Nearly monodisperse Cu-In-S ternary nanocrystals with tunable composition, crystalline structure, and size were synthesized by a hot-injection method using mixed generic precursors. Such ternary nanocrystals with zincblende and wurtzite structure were reported for the first time. This work correlates the crystalline structure of the binary ZnS nanoparticles with those of ternary Cu-In-S nanocrystals, demonstrating the feasibility of making their alloyed or core/shell structure. Furthermore, this work may provide suitable material candidates for low-cost, high-efficiency solar cell fabrication. PMID- 18396870 TI - Boronic acid converters for reactive hydrazide amplifiers: polyphenol sensing in green tea with synthetic pores. AB - Multicomponent sensing in complex matrices with synthetic pores became feasible with the introduction of amplifiers. Amplifiers are defined as molecules that can covalently capture undetectable analytes after enzymatic signal generation and drag them into the pore for transduction. Here, we introduce converters as molecules that can shift the reactivity of amplifiers in situ to capture chemoorthogonal analytes. For this purpose, a series of dialkoxynaphthalene (DAN) and dialkoxyanthracene (DAA) hydrazinoboronic acids was prepared in situ from DAN and DAA hydrazides and formylphenylboronic acids. These converted amplifiers efficiently inactivate synthetic pores with internal naphthalenediimide clamps. This pore inactivation by DAN and DAA hydrazinoboronic acids vanishes in the presence of catechols such as (+)-catechin, presumably because the obtained boronate esters are too large to bind within the synthetic beta-barrel pore or because they prefer to partition into the bilayer membrane. The resulting increase in pore activity with increasing catechol concentration at constant amplifier concentration is shown to be compatible with the sensing of polyphenols in green tea. PMID- 18396871 TI - PEG-detachable polyplex micelles based on disulfide-linked block catiomers as bioresponsive nonviral gene vectors. AB - PEG-based polyplex micelles, which can detach the surrounding PEG chains responsive to the intracellular reducing environment, were developed as nonviral gene vectors. A novel block catiomer, PEG-SS-P[Asp(DET)], was designed as follows: (i) insertion of biocleavable disulfide linkage between PEG and polycation segment to trigger PEG detachment and (ii) a cationic segment based on poly(aspartamide) with a flanking N-(2-aminoethyl)-2-aminoethyl group, P[Asp(DET)], in which the Asp(DET) unit acts as a buffering moiety inducing endosomal escape with minimal cytotoxicity. The polyplex micelles from PEG-SS P[Asp(DET)] and plasmid DNA (pDNA) stably dispersed in an aqueous medium with a narrowly distributed size range of approximately 80 nm due to the formation of hydrophilic PEG palisades while undergoing aggregation by the addition of 10 mM dithiothreitol (DTT) at the stoichiometric charge ratio, indicating the PEG detachment from the micelles through the disulfide cleavage. The PEG-SS P[Asp(DET)] micelles showed both a 1-3 orders of magnitude higher gene transfection efficiency and a more rapid onset of gene expression than PEG P[Asp(DET)] micelles without disulfide linkages, due to much more effective endosomal escape based on the PEG detachment in endosome. These findings suggest that the PEG-SS-P[Asp(DET)] micelle may have promising potential as a nonviral gene vector exerting high transfection with regulated timing and minimal cytotoxicity. PMID- 18396872 TI - Structural, optical, and electrical properties of PbSe nanocrystal solids treated thermally or with simple amines. AB - We describe the structural, optical, and electrical properties of films of spin cast, oleate-capped PbSe nanocrystals that are treated thermally or chemically in solutions of hydrazine, methylamine, or pyridine to produce electronically coupled nanocrystal solids. Postdeposition heat treatments trigger nanocrystal sintering at approximately 200 degrees C, before a substantial fraction of the oleate capping group evaporates or pyrolyzes. The sintered nanocrystal films have a large hole density and are highly conductive. Most of the amine treatments preserve the size of the nanocrystals and remove much of the oleate, decreasing the separation between nanocrystals and yielding conductive films. X-ray scattering, X-ray photoelectron and optical spectroscopy, electron microscopy, and field-effect transistor electrical measurements are used to compare the impact of these chemical treatments. We find that the concentration of amines adsorbed to the NC films is very low in all cases. Treatments in hydrazine in acetonitrile remove only 2-7% of the oleate yet result in high-mobility n-type transistors. In contrast, ethanol-based hydrazine treatments remove 85-90% of the original oleate load. Treatments in pure ethanol strip 20% of the oleate and create conductive p-type transistors. Methylamine- and pyridine-treated films are also p-type. These chemically treated films oxidize rapidly in air to yield, after short air exposures, highly conductive p-type nanocrystal solids. Our results aid in the rational development of solar cells based on colloidal nanocrystal films. PMID- 18396873 TI - Remarkable magneto-optical properties of europium selenide nanoparticles with wide energy gaps. AB - The enhanced magneto-optical properties of nanoscaled lanthanide chalogenide semiconductors which have a wide energy gap were observed at around 500 nm for the first time. The nanoscaled semiconductors, Eu(1-x)Se nanoparticles 1 (cubic shapes) and 2 (spherical shapes), were synthesized by the thermal reduction of Eu(III) ion with organic ligands containing Se atoms. The resulting Eu(1-x)Se nanoparticles were characterized by X-ray diffraction, high-resolution transmission electron microscopy, energy dispersive X-ray spectroscopy, superconducting quantum interference devices magnetometer, and microwave induced plasma atomic emission spectroscopy measurements. The particle grain sizes of 1 and 2 were estimated to be 11 and 20 nm, respectively. The concentration normalized Veldet constants (the magnitude of the Faraday effect) of Eu(1-x)Se nanoparticles were much larger than those of corresponding bulk EuSe and EuS nanoparticles. PMID- 18396874 TI - Access to enantioenriched alpha-amino esters via rhodium-catalyzed 1,4 addition/enantioselective protonation. AB - Conjugate addition of potassium trifluoro(organo)borates 2 to dehydroalanine derivatives 1, mediated by a chiral rhodium catalyst and in situ enantioselective protonation, afforded straightforward access to a variety of protected alpha amino esters 3 with high yields and enantiomeric excesses up to 95%. Among the tested chiral ligands and proton sources, Binap, in combination with guaiacol (2 methoxyphenol), an inexpensive and nontoxic phenol, afforded the highest asymmetric inductions. Organostannanes have also shown to participate in this reaction. By a fine-tuning of the ester moiety, and using Difluorophos as chiral ligand, increased levels of enantioselectivity, generally close to 95%, were achieved. Deuterium labeling experiments revealed, and DFT calculation supported, an unusual mechanism involving a hydride transfer from the amido substituent to the alpha carbon explaining the high levels of enantioselectivity attained in controlling this alpha chiral center. PMID- 18396875 TI - Omni-composites: a new strategy for forming bulk nanostructured materials. AB - This work describes a new route for producing bulk materials from extremely high loading levels of inorganic nanoparticles, approaching 80%, while maintaining optical transparency. More specifically, LaF3 nanoparticles with trifluorovinyl ether (TFVE) ligands were synthesized. Because a single nanoparticle will have many functional ligands attached to its surface, the nanoparticles themselves act as cross-linkers, without the need for a matrix phase for the composite, promoting homogeneity and reduced optical scattering. In order to distinguish these new types of composites from traditional nanocomposites, they are referred to here as "omni"-composites because the material is composed entirely (Lat. omnino) of composite nanoparticles. PMID- 18396876 TI - Pyridine ring opening at room temperature at a rhenium tricarbonyl bipyridine complex. AB - Pyridine ring opening occurs in the reaction of [Re(CO)3(MeIm)(bipy)]OTf with KN(SiMe3)2 followed by double methylation with methyl trifluoromethanesulfonate. Analogues of the neutral product of the initial deprotonation and of the product of the first methylation were isolated by using mesitylimidazole (MesIm) in place of methylimidazole (MeIm) and/or 1,10-phenanthroline (phen) instead of 2,2' bipyridine (bipy). PMID- 18396877 TI - Analysis of protein kinase autophosphorylation using expressed protein ligation and computational modeling. AB - Protein kinases represent a family of enzymes that are critical in cell signaling. One mechanism by which protein kinases are regulated is via autophosphorylation. In the studies described here, we have examined the mechanism of autophosphosphorylation at serine 338 in the regulation of protein kinase A (PKA). Expressed protein ligation allowed for the covalent linkage of an ATP moiety to a Ser mimic at this phosphorylation site. Using a combination of size exclusion chromatography, fluorescence nucleotide binding, kinase measurements, and limited proteolysis assays on this semisynthetic ATP-linked protein, we have obtained unique evidence for an intramolecular autophosphorylation mechanism in PKA regulation. Computational analysis provided a plausible model for a PKA conformation consistent with intramolecular phosphoryl transfer. This approach could be applied to other autoprocessing enzymes by exploiting appropriate transition state analogue motifs in the context of protein semisynthesis. PMID- 18396878 TI - Ligand bonding and dynamics on colloidal nanocrystals at room temperature: the case of alkylamines on CdSe nanocrystals. AB - With CdSe nanocrystals stabilized with very weak ligands (pyridine) as the starting materials, NMR techniques were applied to distinguish the bonded and free alkylamine ligands in an equilibrated adsorption/desorption system for the CdSe-amine nanocrystal-ligand pair. NMR and photoluminescence (PL) measurements were further correlated to identify the linear relationship between PL intensity and the surface ligand coverage of the amine-coated CdSe nanocrystals. For 3.5 nm CdSe nanocrystals and octylamine ligands, the chemical equilibrium constant (K) of the CdSe-amine nanocrystal-ligand adsorption/desorption process was found to be around 50-100, and the corresponding Delta(r)G(o) was calculated as 9.8-11.5 kJ/mol. With a proposed mathematic method, the corresponding chemical kinetic constants for the desorption (kd) and adsorption (ka) processes were measured to be 0.01 s(-1) and 0.5 L mol(-1) s(-1), respectively. K, kd, and ka obtained here are generally 2-4 magnitudes different from those estimated in literature. Analysis indicates that these constants are well consistent with the existing experimental observations. PMID- 18396880 TI - Atom-based thermochemistry: crystal atomization and sublimation enthalpies in linear relationships to molecular atomization enthalpy. AB - In atom-based thermochemistry (ABT), state functions are referenced to free atoms, as opposed to the thermochemical convention of referencing to elements in their standard state. The shift of the reference frame reveals previously unrecognized linear relationships between the standard atomization enthalpies Delta(at)H(o)(g) of gas-phase diatomic and triatomic molecules and Delta(at)H(o)(s) of the corresponding solids for large groups of materials. For 35 alkali and coinage-metal halides, as well as alkali metal hydrides, Delta(at)H(o)(s) = 1.1203 Delta(at)H(o)(g) + 167.0 kJ mol(-1) is found; the standard error is SE = 16.0 kJ mol(-1), and the correlation coefficient is R = 0.9946. The solid coinage-metal monohydrides, CuH(s), AgH(s), and AuH(s), are predicted to be unstable with respect to the formation from the metals and elemental hydrogen by an approximately constant standard enthalpy of formation, Delta(f)H(o)(s) approximately +80 +/- 20 kJ mol(-1). Solid AuF is predicted to be marginally stable, having Delta(f)H(o)(s) = -60 +/- 50 kJ mol(-1) and standard a Gibbs energy of formation Delta(f)G(o)(s) approximately -40 +/- 50 kJ mol (-1). Triatomic alkaline-earth dihalides MX2 obey a similar linear relationship. The combined data of altogether 51 materials obey the relationship Delta(at)H(o)(s) = 1.2593 Delta(at)H(o)(g) + 119.9 kJ mol(-1) with R = 0.9984 and SE = 18.5 kJ mol( 1). The atomization enthalpies per atom of 25 data pairs of diatoms and solids in the groups 14-14, 13-15, and 2-16 are related as Delta(at)H(o)(s) = 2.1015 Delta(at)H(o)(g) + 231.9 kJ mol(-1) with R = 0.9949 and SE = 24.0 kJ mol(-1). Predictions are made for the GeC, GaSb, Hf2, TlN, BeS, MgSe, and MgTe molecules and for the solids SiPb, GePb, SnPb, and the thallium pnictides. Exceptions to the rule, such as SrO and BaO, are rationalized. Standard enthalpies of sublimation, Delta(subl)H(o) = Delta(at)H(o)(s) - Delta(at)H(o)(g), are calculated as a linear function of Delta(at)H(o)(g) profiting from the above linear relationships, and predictions for the Delta(subl)H(o) of the thallium pnictides are given. The validity of the new empirical relationships is limited to substances where at least one of the constituent elements is solid in its standard state. Reasons for the late discovery of such relationships are given, and a meaningful ABT is recommended by using Delta(at)H(o) as an important ordering and reference state function. PMID- 18396879 TI - Rerouting the folding pathway of the Notch ankyrin domain by reshaping the energy landscape. AB - The modular nature of repeat proteins has made them a successful target for protein design. Ankyrin repeat, TPR, and leucine rich repeat domains that have been designed solely on consensus information have been shown to have higher thermostability than their biological counterparts. We have previously shown that we can reshape the energy landscape of a repeat protein by adding multiple C terminal consensus ankyrin repeats to the five N-terminal repeats of the Notch ankyrin domain. Here we explore how the folding mechanism responds to reshaping of the energy landscape. We have used analogous substitutions of a conserved alanine with glycine in each repeat to determine the distribution of structure in the transition state ensembles of constructs containing one (Nank1-5C1) and two consensus (Nank1-5C2) ankyrin repeats. Whereas folding of the wild-type Notch ankyrin domain is slowed by substitutions in its central repeats, (1) folding of Nank1-5C1 and Nank1-5C2 is slowed by substitutions in the C-terminal repeats. Thus, the addition of C-terminal stabilizing repeats shifts the transition state ensemble toward the C-terminal repeats, rerouting the folding pathway of the ankyrin repeat domain. These findings indicate that, for the Notch ankyrin domain, folding pathways are selected based on local energetics. PMID- 18396881 TI - Syntheses of the Stemona alkaloids (+/-)-stenine, (+/-)-neostenine, and (+/-)-13 epineostenine using a stereodivergent Diels-Alder/azido-Schmidt reaction. AB - A tandem Diels-Alder/azido-Schmidt reaction sequence provides rapid access to the core skeleton shared by several Stemona alkaloids including stenine, neostenine, tuberostemonine, and neotuberostemonine. The discovery and evolution of inter- and intramolecular variations of this process and their applications to total syntheses of (+/-)-stenine and (+/-)-neostenine are described. The stereochemical outcome of the reaction depends on both substrate type and reaction conditions, enabling the preparation of both (+/-)-stenine and (+/-)-neostenine from the same diene/dienophile combination. PMID- 18396883 TI - Photomodulated chiral induction in helical azobenzene oligomers. AB - Appending L-alanine to the terminal positions of a helical azobenzene oligomer produced a P helical bias, which increased with oligomer length. Irradiation gave rise to E-->Z isomerization of the terminal azo linkages, which displaced the stereogenic center of L-Ala from the helix backbone and suppressed chiral induction. Theoretical simulations of the CD spectrum of the P helical conformation are in qualitative agreement with the experimental spectra. PMID- 18396884 TI - Distinctive circular dichroism signature for 14-helix-bundle formation by beta peptides. AB - We identify a distinctive circular dichroism (CD) signature for self-assembled 14 helical beta-peptides. Our data show that self-assembly leads to a mimimum at 205 nm, which is distinct from the well-known minimum at 214 nm for a monomeric 14 helix. The onset of assembly is indicated by [theta]205/[theta]214>0.7. Our results will facilitate rapid screening for self-assembling beta-peptides and raise the possibility that far-UV CD will be useful for detecting higher-order structure for other well-folded oligoamide backbones. PMID- 18396885 TI - Picolinoxy group, a new leaving group for anti SN2' selective allylic substitution with aryl anions based on Grignard reagents. AB - The picolinoxy group was found to be an extremely powerful leaving group for allylic substitution with aryl nucleophiles derived from ArMgBr and CuBr*Me2S. The substitution proceeds with anti SN2' pathway and with high chirality transfer. The electron-withdrawing effect of the pyridyl group and chelation to MgBr2 are likely the origin of success. Results suggesting these effects were obtained. PMID- 18396886 TI - Hg(OTf)2-catalyzed arylene cyclization. AB - Novel Hg(OTf) 2-catalyzed arylene cyclization was achieved with highly efficient catalytic turnover (up to 200 times). The reaction takes place via protonation of allylic hydroxyl group by in situ formed TfOH of an organomercuric intermediate to generate a cationic species. Subsequent smooth demercuration regenerates the catalyst. PMID- 18396887 TI - Palladium-catalyzed perarylation of 3-thiophene- and 3-furancarboxylic acids accompanied by C-H bond cleavage and decarboxylation. AB - 3-thiophene- and 3-furancarboxylic acids efficiently undergo perarylation accompanied by cleavage of the three C-H bonds and decarboxylation upon treatment with excess aryl bromides in the presence of a palladium catalyst to give the corresponding tetraarylated products in good yields. PMID- 18396888 TI - Enantioselective synthesis of iridal, the parent molecule of the iridal triterpenoid class. AB - The monocyclic triterpene iridal 1 (parent molecule) is synthesized by an approach that allows access for several representatives of the iridal family as well as diversely substituted analogues. The success of the proposed synthetic plan depends upon the effortless stereoselective establishment of the trans C10/C11 dimethyl relationship in B-ring moiety 7 using a domino-based methodology and the higly efficient Miyaura-Suzuki type sp3-sp2 segment coupling 7 and 8, respectively. PMID- 18396889 TI - Novel one-pot approach to synthesis of indanones through Sb(V)-catalyzed reaction of phenylalkynes with aldehydes. AB - Catalytic SbF5 and the use of EtOH as an additive efficiently converted a mixture of phenylalkynes and aldehydes to indanone compounds in one pot, and the reaction stereoselectively afforded the corresponding 2,3-disubstituted indanones as a single trans-isomer. PMID- 18396890 TI - Fluorescent amino- and thiopyronin dyes. AB - Highly fluorescent rhodamine/rosamine analogues 2 were prepared from ketone 1. PMID- 18396891 TI - Oxidative amination of cuprated pyrimidine and purine derivatives. AB - Using regioselective cuprations (via magnesiations), various primary, secondary and tertiary aminated pyrimidine and purine derivatives were prepared by the oxidative coupling of lithium amidocuprates using chloranil. DNA and RNA units such as aminated uracil or thymine, and adenine, as well as a CDK inhibitor, purvalanol A, were all obtained under mild conditions and satisfactory yields. PMID- 18396892 TI - Rhodium-catalyzed anti selective cross-addition of bis(trimethylsilyl)acetylene to diarylacetylenes via carbon-silicon bond cleavage. AB - The addition of bis(trimethylsilyl)acetylene to diarylacetylenes proceeds efficiently and selectively in a formal anti fashion in the presence of [Rh(OH)(cod)]2/bisphosphine and phenol as catalyst and activator, respectively, accompanied by cleavage of one of the C-Si bonds to produce the corresponding (Z) enynes. The products can further couple with the same or a different diarylacetylene molecule to give rise to (Z,Z)-1,2,5,6-tetraaryl-1,5-hexadien-3 ynes that show relatively strong solid-state fluorescence. PMID- 18396893 TI - Novel bifunctional chiral thiourea catalyzed highly enantioselective aza-Henry reaction. AB - A novel bifunctional chiral thiourea organocatalyst bearing a glycosyl scaffold and a tertiary amino group was synthesized starting from readily available alpha D-glucose. This thiourea was proven to be an effective organocatalyst for the asymmetric aza-Henry reaction between N-Boc imines and nitromethane. The corresponding adducts were obtained in good to excellent yields with excellent enantioselectivities (up to 99.8% ee). In addition, the reaction of nitroethane also proceeded smoothly with excellent enantioselectivity, albeit with low to good diastereoselectivities. PMID- 18396894 TI - Desymmetrization of metallated cyclohexadienes with chiral N-tert-butanesulfinyl imines. AB - This communication describes the desymmetrization of various achiral metallated cyclohexadienes with a series of chiral N-tert-butanesulfinyl imines. Depending on the metal used, either the symmetrical diene (dicyclohexadienyl-zinc) or the desymmetrized diene (cyclohexadienyl-MgCl) is obtained in a good regioselectivity with excellent diastereoselectivity. The products formed should be useful building blocks for natural product synthesis. The symmetrical 1,4-dienes are readily oxidized to the corresponding diarylmethylamine derivatives. PMID- 18396896 TI - Efficient phosphonium-mediated synthesis of 2-amino-1,3,4-oxadiazoles. AB - We present an efficient, room temperature procedure for the preparation of 2 amino-1,3,4-oxadiazoles. Oxadiazol-2-ones can be activated for SNAr substitution using phosphonium reagents (e.g., BOP). This approach provides convenient access to N,N-disubstituted 2-amino-1,3,4-oxadiazoles, which are difficult to prepare using existing synthetic strategies. PMID- 18396895 TI - Second-generation total synthesis of haterumalide NA using B-alkyl Suzuki-Miyaura coupling. AB - Second-generation total synthesis of haterumalide NA, a potent cytotoxic marine macrolide, was achieved by using B-alkyl Suzuki-Miyaura coupling and Nozaki Hiyama-Kishi coupling as key steps (1.2% in 33 steps). Compared to our first generation approach, the second-generation synthesis is much improved in the yield of key intermediate. PMID- 18396898 TI - Inhibitory potency against human acetylcholinesterase and enzymatic hydrolysis of fluorogenic nerve agent mimics by human paraoxonase 1 and squid diisopropyl fluorophosphatase. AB - A wide range of toxic organophosphorus pesticides and nerve agents is effectively hydrolyzed by the structurally related phosphotriesterase enzymes paraoxonase (PON1) from human plasma and diisopropyl fluorophosphatase (DFPase) from the squid Loligo vulgaris. Both enzymes have potential use as medical countermeasures and decontaminants. Enhanced enzymatic activity, stereochemical preference, and substrate variety are still the focus of ongoing research. Derivatives of pesticides and nerve agents bearing a fluorogenic leaving group were introduced for high-throughput screening of mutant libraries recently. We report the inhibitory potency of fluorogenic organophosphorus compounds with three different leaving groups [3-chloro-7-oxy-4-methylcoumarin, 7-oxy-4-methylcoumarin, 7-oxy-4 (trifluoromethyl)coumarin] toward human acetylcholinesterase (AChE) and report kinetic data for the enzymatic hydrolysis of these compounds by PON1 and DFPase. This is the first report of the hydrolysis of a substrate bearing a P-O bond to the leaving group by DFPase (its activity was believed to be restricted to cleavage of P-F and P-CN bonds). The reactivity of the enzymes toward the substrates is explained on the basis of structural reasoning and computational docking studies. We demonstrate that fluorogenic organophosphorus compounds can serve as valuable models for enzyme screening but also show that differences and limitations exist and have to be taken into account. The importance of using protein from human sources to obtain toxicological data for potential in vivo use is highlighted. PMID- 18396900 TI - Synthesis and evaluation of a novel fluorescent photoprobe for imaging matrix metalloproteinases. AB - The measurement of matrix metalloproteinase (MMP) activity in diseases like inflammation, oncogenesis, or atherosclerosis in vivo is highly desirable. Fine tuned pyrimidine-2,4,6-triones (barbiturates) offer nonpeptidyl lead structures for developing imaging agents for specifically visualization of activated MMPs in vivo. The aim of this study was to modify a C-5-disubstituted barbiturate and thus design a highly affine, nonpeptidic, optical MMP inhibitor (MMPI)-ligand for imaging of activated MMPs in vivo. A convergent 10 step synthesis was developed, starting with a malonic ester and (4-bromophenoxy)benzene to generate 5-bromo pyrimidine-2,4,6-trione as the key intermediate. To minimize the interactions between activated MMPs and the dye of the conjugate 6, a PEGylated piperazine derivative was used as a spacer and an azide as a protected amino function. After linking both building blocks, reducing the azide ( Staudinger reaction) and labeling with Cy 5.5, we obtained the nonhydroxamate MMP inhibitor 6 with high affinity (IC 50-value: 48 nM for MMP-2) measured in a fluorogenic assay using commercially available MMP-substrates and the purified enzyme. Zymography revealed an efficient blocking of enzyme activity of purified MMP-2 and MMP-9 and of MMP-containing cell supernatants (HT-1080), (A-673) using the PEGylated barbiturate 5. Fluorescence microscopy studies using a highly (A-673) and a moderate (HT-1080) MMP-2 secreting cell line showed efficient binding of the Cy 5.5 labeled tracer 6 to the MMP-2 positive cells while MMP-2 negative cells (MCF 7) did not bind. Therefore, this new barbiturate-based MMP-probe has a high affinity and specificity toward MMP-2 and -9 and is thus a promising candidate for sensitive MMP detection in vivo. PMID- 18396901 TI - Proteomic profiling of nonenzymatically glycated proteins in human plasma and erythrocyte membranes. AB - Nonenzymatic glycation of peptides and proteins by d-glucose has important implications in the pathogenesis of diabetes mellitus, particularly in the development of diabetic complications. In this work, we report the first proteomics-based characterization of nonenzymatically glycated proteins in human plasma and erythrocyte membranes from individuals with normal glucose tolerance, impaired glucose tolerance, and type 2 diabetes mellitus. Phenylboronate affinity chromatography was used to enrich glycated proteins and glycated tryptic peptides from both human plasma and erythrocyte membranes. The enriched peptides were subsequently analyzed by liquid chromatography coupled with electron transfer dissociation-tandem mass spectrometry, resulting in the confident identification of 76 and 31 proteins from human plasma and erythrocyte membranes, respectively. Although most of the glycated proteins could be identified in samples from individuals with normal glucose tolerance, slightly higher numbers of glycated proteins and more glycation sites were identified in samples from individuals with impaired glucose tolerance and type 2 diabetes mellitus. PMID- 18396902 TI - Proteomic screening of a cell line model of esophageal carcinogenesis identifies cathepsin D and aldo-keto reductase 1C2 and 1B10 dysregulation in Barrett's esophagus and esophageal adenocarcinoma. AB - Esophageal adenocarcinoma (EA) incidence is increasing rapidly and is associated with a poor prognosis. Identifying biomarkers of disease development and progression would be invaluable tools to inform clinical practice. Two dimensional polyacrylamide gel electrophoresis was used to screen 10 esophageal cell lines representing distinct stages in the development of esophageal cancer. Thirty-three proteins were identified by MALDI-TOF-MS which demonstrated differences in expression across the cell lines. Western blotting and qRT-PCR confirmed increased cathepsin D and aldo-keto reductases 1C2 and 1B10 expression in metaplastic and dysplastic cell lines. Expression of these proteins was further assessed in esophageal epithelium from patients with nonerosive (NERD) and erosive gastro-esophageal reflux disease, Barrett's esophagus (BE) and EA. When compared with normal epithelium of NERD patients, (i) cathepsin D mRNA levels demonstrated a stepwise increase in expression (p<0.05) in erosive, metaplastic and EA tissue; (ii) AKR1B10 expression increased (p<0.05) 3- and 9 fold in erosive and Barrett's epithelium, respectively; and (iii) AKR1C2 levels increased (p<0.05) in erosive and Barrett's epithelium, but were reduced (p<0.05) in EA. These proteins may contribute to disease development via effects on apoptosis, transport of bile acids and retinoid metabolism and should be considered as candidates for further mechanistic and clinical investigations. PMID- 18396903 TI - Subinhibitory concentrations of antibiotics induce phenazine production in a marine Streptomyces sp. AB - Subinhibitory concentrations of antibiotics were found to enhance and modulate the production of new phenazines, streptophenazines A-H, in a marine Streptomyces isolate. The streptophenazines differ in length and substitution of an alkyl chain. The pattern of metabolites formed depends on the antibiotic used. In the presence of tetracycline, streptophenazines F and G were induced and the production of streptophenazines A-D was increased. When using bacitracin, mainly streptophenazine H was produced. Streptophenazines C and H showed moderate activity against Bacillus subtilis, while streptophenazine C was also active against Staphylococcus lentus. PMID- 18396904 TI - Lignan glycosides from Neoalsomitra integrifoliola. AB - Five new lignan glycosides, (8R*,7' S*,8' R*)-5,5'-dimethoxy-7-oxolariciresinol 9'-O-beta-D-xylopyranoside (1), (7S,8R)-dihydrodehydrodiconiferyl alcohol 9-O beta-D-apiofuranosyl-(1-->6)-O-beta-D-glucopyranoside (2), (7S*,8R*,7' S*,8' R*) 4,4'-dimethoxyhuazhongilexin 9-O-beta-D-xylopyranoside (4), (7S*,8R*,7' S*,8' R*) 4,4'-dimethoxyhuazhongilexin 9-O-alpha-L-arabinopyranoside (5), and (7S*,8R*,7' S*,8' R*)-huazhongilexin 9-O-(2-feruloyl)-beta-D-xylopyranoside (6), together with four known compounds, (7R,8S)-dihydrodehydrodiconiferyl alcohol 9-O-beta-D glucopyranoside (3), the 9-O-beta-D-xylopyranoside of icariol A(2) (7), huazhongilexin (8), and nudiposide (9), were isolated from the leaves of Neoalsomitra integrifoliola. Their structures were determined by spectroscopic analysis and chemical methods. Compounds 2, 3, 8, and 9 showed weak anti inflammatory activities. PMID- 18396905 TI - Periglaucines A-D, anti-HBV and -HIV-1 alkaloids from Pericampylus glaucus. AB - Four new hasubanane-type alkaloids, periglaucines A-D (1-4), and three known alkaloids, norruffscine (5), (-)-8-oxotetrahydropalmatine (6), and (-)-8 oxocanadine (7), were isolated from the aerial parts of Pericampylus glaucus. Their structures were elucidated on the basis of extensive NMR and EIMS data, and that of periglaucine A (1) was confirmed by single-crystal X-ray diffraction. Alkaloids 1-4 inhibited hepatitis B virus (HBV) surface antigen (HBsAg) secretion in Hep G2.2.15 cells. (-)-8-Oxotetrahydropalmatine (6) possessed a high selectivity index (SI = 22.4) for HBsAg secretion of the Hep G2.2.15 cell line with an IC(50) value of 0.14 mM. Norruffscine (5) and (-)-8 oxotetrahydropalmatine (6) exhibited inhibitory activity against human immunodeficiency virus (HIV-1) with EC(50) values of 10.9 and 14.1 microM in C8166 cells (SI = 45.7 and 18.8), respectively. PMID- 18396906 TI - Anion and carboxylic acid binding to monotopic and ditopic amidopyridine macrocycles. AB - Binding of inorganic anions, carboxylic acids, and tetraalkylammonium carboxylates by macrocyclic compounds of different size was studied by NMR in DMSO-d6. It has been shown that at least a 15-membered ring is necessary for successful recognition of fluoride. Larger macrocycles were shown to bind HSO4( ), H2PO4(-), Cl(-), and carboxylic acid salts. Effects of binding topicity are discussed. The 30-membered macrocycles 4 and 4m selectively bind substrates that are size- and shape-complementary: maximum binding is observed for dicarboxylic acids and dicarboxylates with four-carbon chains, and the binding constant for association of fumaric acid and 4 is ca. 5 orders of magnitude higher than that of maleic acid. The 30-membered macrocycle 4m showed selectivity toward alpha ketocarboxylic acids. Secondary amino groups were not crucial for binding of fluoride to the macrocycles; however, they proved to be very important for selectivity and strength of carboxylic acid binding. The X-ray structure of the adduct of 4 and nitrobenzoic acid confirmed the guest H-bonding with both the amide and the secondary amino groups of the 30-membered macrocyclic host. PMID- 18396907 TI - Kinetic study of electrochemically induced michael reactions of o-quinones with Meldrum's acid derivatives. Synthesis of highly oxygenated catechols. AB - Electrochemical oxidation of catechols has been studied in the presence of Meldrum's acid derivatives as nucleophiles in aqueous solution, by means of cyclic voltammetry and controlled-potential coulometry. Catechols in the Michael addition reaction react with Meldrum's acids to form adducts that can undergo electrooxidation. Such products were obtained in good yields as confirmed by controlled potential electrosynthesis. Such products can be generated in aqueous solutions by means of electrosynthesis, using a carbon electrode in an undivided cell. Furthermore, the homogeneous rate constants of the chemical reaction interposed between electron transfers were estimated by comparing the experimental cyclic voltammetric curves with the digitally simulated ones. PMID- 18396908 TI - Indium-copper and indium-silver mediated barbier-grignard-type alkylation reaction of aldehydes using unactivated alkyl halides in water. AB - An efficient method has been developed for the Barbier-Grignard-type alkylation reaction of aldehydes (including aliphatic version) using unactivated alkyl halides in water in the presence of an In/CuI/I(2) or In/AgI/I(2) system. The reactions proceeded more efficiently in water than in organic solvent. In, CuI or AgI, and I(2) were all essential for the efficient progress of the reactions. A radical-type reaction mechanism was studied and proposed by using 4-pentenal as substrate. PMID- 18396909 TI - Organocatalytic and enantioselective synthesis of beta-(hydroxyalkyl)-gamma butyrolactones. AB - Organocatalytic cross-aldol reaction of methyl 4-oxobutyrate (2) and a variety of aldehydes 3 followed by reduction with NaBH(4) has provided a one-pot, general and efficient method for the synthesis of 4-(hydroxyalkyl)-gamma-butyrolactones 1 with high diastereo-(dr > 24:1) and enantioselectivity (ee > 99%). PMID- 18396910 TI - Nonbonded, attractive cation-pi interactions in azide-mediated asymmetric ring expansion reactions. AB - The influence of attractive, nonbonded interactions on the reactions of 1,2- and 1,3-hydroxyalkyl azides with ketones has been investigated through experimental and computational means. A series of 1,3-hydroxyalkyl azides bearing electronically tuned aromatic groups at the 2 position were prepared and reacted along with several derivatives designed to conformationally restrict the rotational orientation of the aromatic substituent. These studies showed that a cation-pi interaction between an aryl moiety and an N2(+) leaving group plays a role in determining the stereoselectivity of these reactions. A series of ab initio calculations supported this hypothesis. A computational and experimental analysis suggested a primarily steric model for the analogous reactions of substituted 2-azido-1-ethanol analogues. PMID- 18396911 TI - Enhanced secretion of heterologous proteins from yeast by overexpression of ribosomal subunit RPP0. AB - Previously, we have shown that single-gene overexpression of five yeast genes (CCW12, CWP2, ERO1, RPP0, and SED1) promoted increased secretion levels of several single-chain antibody fragments and single-chain T-cell receptors from Saccharomyces cerevisiae (Wentz, A. E.; Shusta, E. V. Appl. Environ. Microbiol. 2007, 73, (4), 1189-1198). In this study, several proteins possessing different protein folds were secreted from yeast overexpressing each of the five genes to determine the generality of the secretion enhancers. Only one gene encoding a ribosomal subunit (RPP0) enhanced secretion levels for multiple proteins: a single-chain antibody (the 4-4-20 anti-fluorescein scFv) and green fluorescent protein (GFP). Protein induction time-course experiments revealed increased secretion with RPP0 overexpression for 4-4-20 as early as 40 h post-induction. Effects on GFP secretion levels were not evident until late induction times where overexpression of RPP0 limited post-secretion protein loss, but absolute yields did not exceed those observed at earlier induction times. The effects of RPP0 overexpression on secreted protein yields did not appear to directly involve ribosome function, but instead RPP0 overexpression indirectly regulated acidification of the yeast medium by preventing upregulation of the yeast plasma membrane H+-ATPase gene, PMA1. Combining RPP0 overexpression with nutrient supplementation stimulated additional protein secretion for the 4-4-20 scFv with higher per cell secretion that corresponded to 6-fold increases in volumetric yield. PMID- 18396912 TI - Enhancement of mussel adhesive protein production in Escherichia coli by co expression of bacterial hemoglobin. AB - Mussel adhesive proteins (MAPs) have been considered as potential underwater and medical bioadhesives. Previously, we reported a functional expression of recombinant MAP hybrid fp-151, which is a fusion protein with six type 1 (fp-1) decapeptide repeats at each type 5 (fp-5) terminus, with practical properties in Escherichia coli. In the present work, we introduced the Vitreoscilla hemoglobin (VHb) co-expression strategy to enhance the production levels of hybrid fp-151 since VHb has been successfully used for efficient oxygen utilization in several expression systems, including E. coli. In both batch-type flask and fed-batch type bioreactor cultures, we found that co-expression of VHb conferred higher cell growth and hybrid fp-151 production. Its positive effects were significantly increased in high cell density bioreactor cultures as the microaerobic environment was more quickly and severely formed. We obtained a approximately 1.9 fold higher (approximately 1 g/L) production of MAP fp-151 from VHb co-expressing cells in fed-batch bioreactor cultures as compared to that from VHb non expressing cells. Collectively and regardless of the culture type, VHb co expression strategy was successful in enhancing the production of recombinant mussel adhesive proteins in the E. coli expression system. PMID- 18396913 TI - Cloning and characterization of long-chain fatty alcohol oxidase LjFAO1 in lotus japonicus. AB - The Lotus japonicus EST database was searched against Arabidopsis thaliana AtFAO3, a full-length cDNA that encodes a membrane-bound, flavin-containing, hydrogen peroxide generating, long-chain fatty alcohol oxidase. One EST fragment was detected, and the corresponding full-length cDNA was obtained by screening a cDNA library of L. japonicus. The LjFAO1 genomic DNA was amplified by PCR, to give a product 3.6 kb in length. Comparison between the LjFAO1 cDNA and genomic DNA revealed that the LjFAO1 contains 3 exons and 2 introns. RT-PCR analysis showed that the LjFAO1 was expressed in the whole plant, with the highest expression level in the apex and the lowest expression level in the siliques. The LjFAO1 gene was down-regulated by cold stress in both the apex and the cotelydon of the 8-day old seedlings, the first time that a long-chain alcohol oxidase has been shown to respond directly to stress. The full length cDNA and a C-terminal truncated version were overexpressed in Escherichia coli. The full length version of LjFAO1 exhibited long-chain fatty alcohol oxidase activity and was subsequently purified by Ni-NTA chromatography. The active LjFAO1 protein showed substrate specificities toward 1-dodecanol, 1-hexadecanol, and 1,16 hexadecanediol with Km values 59.6 +/- 14.8 (microM), 40.9 +/- 8.2 (microM) and 19.4 +/- 1.5 (microM), respectively, suggesting apparent differences in substrate preferences with AtFAO3. PMID- 18396914 TI - Two-dimensional method for time aligning liquid chromatography-mass spectrometry data. AB - We describe a new time alignment method that takes advantage of both dimensions of LC-MS data to resolve ambiguities in peak matching while remaining computationally efficient. This approach, Warp2D, combines peak extraction with a two-dimensional correlation function to provide a reliable alignment scoring function that is insensitive to spurious peaks and background noise. One dimensional alignment methods are often based on the total-ion-current elution profile of the spectrum and are unable to distinguish peaks of different masses. Our approach uses one-dimensional alignment in time, but with a scoring function derived from the overlap of peaks in two dimensions, thereby combining the specificity of two-dimensional methods with the computational performance of one dimensional methods. The peaks are approximated as two-dimensional Gaussians of varying width. This approximation allows peak overlap (the measure of alignment quality) to be calculated analytically, without computationally intensive numerical integration in two dimensions. To demonstrate the general applicability of Warp2D, we chose a variety of complex samples that have substantial biological and analytical variability, including human serum and urine. We show that Warp2D works well with these diverse sample sets and with minimal tuning of parameters, based on the reduced standard deviation of peak elution times after warping. The combination of high computational speed, robustness with complex samples, and lack of need for detailed tuning makes this alignment method well suited to high throughput LC-MS studies. PMID- 18396916 TI - Po(IV) hydration: a quantum chemical study. AB - This work presents a theoretical study on the hydration of Po(IV) in solution. Three points have been addressed: (i) the level of calculation needed to properly describe the system under study, (ii) the hydration number of Po(IV), and (iii) the nature of the polonium-water bonding. The condensed medium effects have been included by means of a continuum solvation model, thus different [Po(H(2)O)(n)](4+) hydrates were embedded in a cavity surrounded by a polarizable dielectric medium. Among the quantum-mechanical calculation levels here considered, the MPW1PW91 functional was shown to be the most suitable, allowing a proper description of the Po-H(2)O interactions at affordable cost. The hydration number of Po(IV) was found to be between 8 and 9. This value is ruled by a dynamic equilibrium involving the octa- and ennea-hydrates, although the 7-fold coordination cannot be completely excluded. The hydration free energy of Po(IV) is estimated to be around -1480 kcal/mol. The Po-H(2)O bonding is dominated by strong electrostatic contributions although a small covalent contribution is responsible for the peculiar arrangement adopted by the smaller hydrates (n < or = 5). A natural bond order (NBO) analysis of the hydrate wave functions shows that the covalent bond involves the empty 6p orbitals of the polonium ion and one lone pair on the oxygen atom of the water molecule. A parallel investigation to the hydrate study, where the polonium ion was replaced by a tetravalent point charge plus a repulsion potential, was carried out. These results allowed a detailed examination of the electrostatic and nonelectrostatic contributions to the polonium hydrate formation. PMID- 18396915 TI - Rapidly alternating transmission mode electron-transfer dissociation and collisional activation for the characterization of polypeptide ions. AB - Cation transmission/electron-transfer reagent anion storage mode electron transfer ion/ion reactions and beam-type collisional activation of the polypeptide ions are performed in rapid succession in the high-pressure collision cell (Q2) of a quadrupole/time-of-flight tandem mass spectrometer (QqTOF), where the electron-transfer reagent anions are accumulated. Duty cycles for both electron-transfer dissociation (ETD) and collision-induced dissociation (CID) experiments are improved relative to ion trapping approaches since there are no discrete ion storage and reaction steps for ETD experiments and no discrete ion storage step and frequency tuning for CID experiments. For this technique, moderately high resolution and mass accuracy are also obtained due to mass analysis via the TOF analyzer. This relatively simple approach has been demonstrated with a triply charged tryptic peptide, a triply charged tryptic phosphopeptide, and a triply charged tryptic N-linked glycopeptide. For the tryptic peptide, the sequence is identified with more certainty than would be available from a single method alone due to the complementary information provided by these two dissociation methods. Because of the complementary information derived from both ETD and CID dissociation methods, peptide sequence and post-translational modification (PTM) sites for the phosphopeptide are identified. This combined ETD and CID approach is particularly useful for characterizing glycopeptides because ETD generates information about both peptide sequence and locations of the glycosylation sites, whereas CID provides information about the glycan structure. PMID- 18396917 TI - Operation of the proton wire in green fluorescent protein. A quantum dynamics simulation. AB - A nuclear quantum dynamical simulation of the proton shuttle operating in the green fluorescent protein has been carried out on a high-quality, high dimensionality potential energy surface describing the photoactive pipi* excited state, and including motion of both the three protons and of the donor and acceptor atoms of the hydrogen bonds in a closed proton wire. The results of the simulations show that proton transfer along the wire is essentially concerted, synchronous, and very fast, with a substantial amount of the green fluorescent species forming within several tens of femtoseconds. In this regard, analysis of the population of the fluorescent species indicates that at least two dynamical regimes are present for its formation. Within the first hundreds of femtoseconds, dynamics is very fast and impulsive. Later on, a slower pace of formation appears. It is discussed that the two largest decay times for the protonated chromophore reported experimentally (Chattoraj, M.; King, B. A.; Bublitz, G. U.; Boxer, S. G. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 8362-8367) might correspond to some irreversible process occurring after formation of the fluorescent species, rather than to cleavage of the chromophore's phenolic O-H bond. PMID- 18396918 TI - Segmentation of verb forms in preverbal infants. AB - It has been observed that children's early vocabulary is dominated by nouns, with verbs being much delayed. The current study investigated if this delay is related to infants' failure to segment verb forms. Using a preferential looking procedure, French-learning preverbal infants were tested on novel verbs segmentation. Infants at the onset of vocabulary learning (11-month-olds) succeeded in segmenting the targets: they listened longer to test sentences containing previously familiarized verbs versus those containing nonfamiliarized verbs, suggesting that the delay in verb learning is not due to segmentation difficulty. Semantic and syntactic complexities of verbs could be among the underlying factors. PMID- 18396919 TI - Tissue mimicking materials for dental ultrasound. AB - While acoustic tissue mimicking materials have been explored for a variety of soft and hard biological tissues, no dental hard tissue mimicking materials have been characterized. Tooth phantoms are necessary to better understand acoustic phenomenology within the tooth environment and to accelerate the advancement of dental ultrasound imaging systems. In this study, soda lime glass and dental composite were explored as surrogates for human enamel and dentin, respectively, in terms of compressional velocity, attenuation, and acoustic impedance. The results suggest that a tooth phantom consisting of glass and composite can effectively mimic the acoustic behavior of a natural human tooth. PMID- 18396920 TI - The effect of stimulus range on two-interval frequency discrimination. AB - It has traditionally been thought that performance in two-interval frequency discrimination tasks decreases as the range over which the standard tone varies is increased. Recent empirical evidence and a reexamination of previous results suggest that this may not be the case. The present experiment found that performance was significantly better when the standard roved over a wide range (1500 Hz) than a narrow range (30 Hz). This pattern cannot readily be accommodated by traditional models of frequency discrimination based on memory or attention, but may be explicable in terms of neural plasticity and the formation of perceptual anchors. PMID- 18396921 TI - Rollover effect of signal level on vowel formant discrimination. AB - The goal of this study was to measure the ability of normal-hearing listeners to discriminate formant frequency for vowels in isolation and sentences at three signal levels. Results showed significant elevation in formant thresholds as formant frequency and linguistic context increased. The signal level indicated a rollover effect, especially for F2, in which formant thresholds at 85 dB SPL were lower than thresholds at 70 or 100 dB SPL in both isolated vowels and sentences. This rollover level effect could be due to reduced frequency selectivity and forward/backward masking in sentence at high signal levels for normal-hearing listeners. PMID- 18396922 TI - Effect of spectral resolution on the intelligibility of ideal binary masked speech. AB - Most binary-mask studies assume a fine time-frequency representation of the signal that may not be available in some applications (e.g., cochlear implants). This study assesses the effect of spectral resolution on intelligibility of ideal binary masked speech. In Experiment 1, speech corrupted in noise at -5 to 5 dB signal-to-noise ratio (SNR) was filtered into 6-32 channels and synthesized using the ideal binary mask. Results with normal-hearing listeners indicated substantial improvements in intelligibility with 24-32 channels, particularly in 5 dB SNR. Results from Experiment 2 indicated that having access to the ideal binary mask in the F1/F2 region is sufficient for good performance. PMID- 18396923 TI - Interpretations on principal components analysis of head-related impulse responses in the median plane. AB - A principal components analysis of the median-plane head-related impulse responses (HRIRs) in the CIPIC HRTF database reveals that the individual HRIRs can be reconstructed by a linear combination of 12 principal components (PCs) within 5% of error in the least-squares sense. The PCs include the intersubject and interelevation variations in the median-plane HRIRs. Each PC provides sound cues for the front-back discrimination and/or the vertical perception. There exist common systematic elevation dependencies in the weights of lower-numbered PCs which contribute to the pinna/head diffractions, whereas the elevation dependencies in the weights of higher-numbered PCs are different from subject to subject. PMID- 18396924 TI - Tactile enhancement of auditory and visual speech perception in untrained perceivers. AB - A single pool of untrained subjects was tested for interactions across two bimodal perception conditions: audio-tactile, in which subjects heard and felt speech, and visual-tactile, in which subjects saw and felt speech. Identifications of English obstruent consonants were compared in bimodal and no tactile baseline conditions. Results indicate that tactile information enhances speech perception by about 10 percent, regardless of which other mode (auditory or visual) is active. However, within-subject analysis indicates that individual subjects who benefit more from tactile information in one cross-modal condition tend to benefit less from tactile information in the other. PMID- 18396925 TI - Dial A440 for absolute pitch: absolute pitch memory by non-absolute pitch possessors. AB - Listeners without absolute (or "perfect") pitch have difficulty identifying or producing isolated musical pitches from memory. Instead, they process the relative pattern of pitches, which remains invariant across pitch transposition. Musically untrained non-absolute pitch possessors demonstrated absolute pitch memory for the telephone dial tone, a stimulus that is always heard at the same absolute frequency. Listeners accurately classified pitch-shifted versions of the dial tone as "normal," "higher than normal" or "lower than normal." However, the role of relative pitch processing was also evident, in that listeners' pitch judgments were also sensitive to the frequency range of stimuli. PMID- 18396926 TI - Simultaneous estimation of attenuation and structure parameters of aggregated red blood cells from backscatter measurements. AB - The analysis of the ultrasonic frequency-dependent backscatter coefficient of aggregating red blood cells reveals information about blood structural properties. The difficulty in applying this technique in vivo is due to the frequency-dependent attenuation caused by intervening tissue layers that distorts the spectral content of backscattering properties from blood microstructures. An optimization method is proposed to simultaneously estimate tissue attenuation and blood structure factor. With in vitro experiments, the method gave satisfactory estimates with relative errors below 22% for attenuations between 0.101 and 0.317 dBcmMHz, signal-to-noise ratios>28 dB and kR<2.7 (k being the wave number and R the aggregate radius). PMID- 18396927 TI - Illusory spectrotemporal ripples created with binaurally correlated noise. AB - Binaural disparities are the primary acoustic cues employed in sound localization tasks. However, the degree of binaural correlation in a sound serves as a complementary cue for detecting competing sound sources [J. F. Culling, H. S. Colburn, and M. Spurchise, "Interaural correlation sensitivity," J. Acoust. Soc. Am. 110(2), 1020-1029 (2001) and L. R. Bernstein and C. Trahiotis, "On the use of the normalized correlation as an index of interaural envelope correlation," J. Acoust. Soc. Am. 100, 1754-1763 (1996)]. Here a random chord stereogram (RCS) sound is developed that produces a salient pop-out illusion of a slowly varying ripple sound [T. Chi et al., "Spectro-temporal modulation transfer functions and speech intelligibility," J. Acoust. Soc. Am. 106(5), 2719-2732 (1999)], even though the left and right ear sounds alone consist of noise-like random modulations. The quality and resolution of this percept is systematically controlled by adjusting the spectrotemporal correlation pattern between the left and right sounds. The prominence and limited time-frequency resolution for resolving the RCS suggests that envelope correlations are a dominant binaural cue for grouping acoustic objects. PMID- 18396928 TI - The low-frequency sound speed of fluid-like gas-bearing sediments. AB - The low-frequency sound speed in a fluid-like kaolinite sediment containing air bubbles was measured using an acoustic resonator technique and found to be 114 ms with negligible dispersion between 100 and 400 Hz. The sediment's void fraction and bubble size distribution was determined from volumetric images obtained from x-ray computed tomography scans. A simplified version of Wood's effective medium model, which is dependent only upon the ambient pressure, the void fraction, the sediment's bulk mass density, and the assumption that all the bubbles are smaller than resonance size at the highest frequency of interest, described the measured sound speed. PMID- 18396987 TI - Comment on "silent research vessels are not quiet" [J. Acoust. Soc. Am. 121, EL145-EL150]. AB - The recent paper by Ona et al. [J. Acoust. Soc. Am. 121, EL145-EL150] compared avoidance reactions by herring (Clupea harengus) to a traditional and a "silent" research vessel. Surprisingly, the latter evoked the strongest avoidance, leading to the conclusion that "candidate stimuli for vessel avoidance remain obscure." In this Comment, it is emphasized that the otolith organs in fish are linear acceleration detectors with extreme sensitivity to infrasonic particle acceleration. Near-field particle motions generated by a moving hull are mainly in the infrasonic range, and infrasound is particularly potent in evoking directional avoidance responses in several species of fish. The stimuli initiating vessel avoidance may thus include infrasonic particle acceleration. PMID- 18396988 TI - Target detection and location with ambient noise. AB - By placing a vertical array in an ambient noise field and forming an upward and a downward beam one obtains two time series which can be cross correlated to reveal a subbottom profile of the seabed [Siderius et al., J. Acoust. Soc. Am. 120, 1315 1323 (2006)]. Here the cross-correlation approach is applied to the location in range and bearing of a point target. An experiment was designed using floats and weights mounted (and dismounted) on the same cable as the vertical array. Careful measurements were made of the location of all likely floats, ballast weights, array terminations, and so on. After suitable coherent averaging, peaks were seen at delays (correlation offsets) agreeing with the reflector positions and were shown to be absent when reflectors were removed. A trivial extension of the theory developed in Harrison and Siderius [J. Acoust. Soc. Am. 123, 1282-1296 (2008)] is used to explain the rough amplitudes of the reflections. The approach differs from "acoustic daylight" principally in having a capability to determine a target range. PMID- 18396989 TI - The performance of active control of random noise in cars. AB - This letter investigates the effectiveness of various configurations of reference sensors for feedforward active control systems in vehicles using unconstrained frequency domain optimization. The results from a model problem are based on a fully coupled analysis between the vibration of a car panel and an enclosed acoustic field. These suggest that with correct locations, only a small number of microphones or accelerometers are needed to give good overall performance, despite many uncorrelated primary disturbances being present. Similar results are predicted from road test data. PMID- 18396990 TI - A comparison of statistically optimized near field acoustic holography using single layer pressure-velocity measurements and using double layer pressure measurements. AB - Statistically optimized near field acoustic holography (SONAH) is usually based on the assumption that all sources are on one side of the measurement plane whereas the other side is source free. An extension of the SONAH procedure based on measurement with an array of pressure-velocity probes has recently been suggested. An alternative method uses a double layer array of pressure transducers. Both methods make it possible to distinguish between sources on the two sides of the array and thus suppress the influence of extraneous noise and reflections coming from the "wrong" side. This letter compares the two methods. PMID- 18396991 TI - The decay of pitch memory during rehearsal. AB - The present study investigates the decay of pitch memory over time. In a delayed pitch comparison paradigm, participants had to memorize the pitch of a Shepard tone, with silent, overt, or without any rehearsal. During overt rehearsal, recordings of the rehearsing were effectuated. Performance was best for silent rehearsal and worst for overt rehearsal. The differences, although partially significant, were not marked. The voice pitch during overt rehearsal was compatible with a random walk model, providing a possible explanation of why rehearsal does not improve the retention of the pitch trace. PMID- 18396992 TI - Detection-theoretic analysis of the observer-based psychophysical procedure. AB - The observer-based procedure, used in research applications to measure the hearing sensitivity of young infants, is analyzed within the framework of a two stage (infant-judge) detection model involving a minimum number of assumptions. The model is taken to develop convergence theorems useful for estimating the number of judges and/or trials required to achieve a desired level of accuracy using the procedure. The model is also used to consider ways in which bias in estimates, known to be associated with the procedure, might be evaluated and reduced. Finally, a method is proposed by which the results of the analysis and different procedural variations designed to improve estimates can be evaluated empirically. PMID- 18396993 TI - A simulation tool for brassiness studies. AB - A frequency-domain numerical model of brass instrument sound production is proposed as a tool to predict their brassiness, defined as the rate of spectral enrichment with increasing dynamic level. It is based on generalized Burger's equations dedicated to weakly nonlinear wave propagation in nonuniform ducts, and is an extension of previous work by Menguy and Gilbert [Acta Acustica 86, 798-810 (2000)], initially limited to short cylindrical tubes. The relevance of the present tool is evaluated by carrying out simulations over distances longer than typical shock formation distances, and by doing preliminary simulations of periodic regimes in a typical brass trombone bore geometry. PMID- 18396994 TI - Behavioral avoidance threshold level of a harbor porpoise (Phocoena phocoena) for a continuous 50 kHz pure tone. AB - The use of ultrasonic sounds in alarms for gillnets may be advantageous, but the deterring effects of ultrasound on porpoises are not well understood. Therefore a harbor porpoise in a large floating pen was subjected to a continuous 50 kHz pure tone with a source level of 122+/-3 dB (re 1 microPa, rms). When the test signal was switched on during test periods, the animal moved away from the sound source. Its respiration rate was similar to that during baseline periods, when the sound was switched off. The behavior of the porpoise was related to the sound pressure level distribution in the pen. The sound level at the animal's average swimming location during the test periods was approximately 107+/-3 dB (re 1 microPa, rms). The avoidance threshold sound pressure level for a continuous 50 kHz pure tone for this porpoise, in the context of this study, is estimated to be 108+/-3 dB (re 1 microPa, rms). This study demonstrates that porpoises may be deterred from an area by high frequency sounds that are not typically audible to fish and pinnipeds and would be less likely masked by ambient noise. PMID- 18396995 TI - Modeling the acoustic radiation force in microfluidic chambers. AB - A procedure is demonstrated to quantitatively evaluate the acoustic radiation forces in microfluidic particle manipulation chambers. Typical estimates of the acoustic pressure and the acoustic radiation force are based on an analytical solution for a simple one-dimensional standing wave pattern. The complexities of a typical microfluidic channel limit the usefulness of this approach. By leveraging finite elements, and a generalized equation for the acoustic radiation force, channel designs can be investigated in two and three dimensions. Calculations and experimental observations in this report and the literature, confirm these claims. PMID- 18396997 TI - On the sound field of a resilient disk in free space. AB - Radiation characteristics are calculated for a circular planar sound source in free space with a uniform surface pressure distribution, which can be regarded as a freely suspended membrane with zero mass and stiffness. This idealized dipole source is shown to have closed form solutions for its far-field pressure response and radiation admittance. The latter is found to have a simple mathematical relationship with the radiation impedance of a rigid piston in an infinite baffle. Also, a single expansion is derived for the near-field pressure field, which degenerates to a closed form solution on the axis of symmetry. From the normal gradient of the surface pressure, the surface velocity is calculated. The near-field expression is then generalized to an arbitrary surface pressure distribution. It is shown how this can be used as a simplified solution for a rigid disk in free space or a more realistic sound source such as pre-tensioned membrane in free space with non-zero mass and a clamped rim. PMID- 18396996 TI - Low-frequency sound transmission through a gas-liquid interface. AB - Typically, sound speed in gases is smaller and mass density is much smaller than in liquids, resulting in a very strong acoustic impedance contrast at a gas liquid interface. Sound transmission through a boundary with a strong impedance contrast is normally very weak. This paper studies the power output of localized sound sources and acoustic power fluxes through a plane gas-liquid interface in a layered medium. It is shown that, for low-frequency sound, a phenomenon of anomalous transparency can occur where most of the acoustic power generated by a source in a liquid half-space can be radiated into a gas half-space. The main physical mechanism responsible for anomalous transparency is found to be an acoustic power transfer by inhomogeneous (evanescent) waves in the plane-wave decomposition of the acoustic field in the liquid. The effects of a liquid's stratification and of guided sound propagation in the liquid on the anomalous transparency of the gas-liquid interface are considered. Geophysical and biological implications of anomalous transparency of water-air interface to infrasound are indicated. PMID- 18396998 TI - An acoustic intensity-based method for reconstruction of radiated fields. AB - An acoustic intensity-based method is proposed for the reconstruction of acoustic radiation pressure. Unlike the traditional inverse acoustic methods, the proposed method includes the acoustic pressure gradient as an input in addition to its simultaneous, co-located acoustic pressure in a radiated field. As a result, the reconstruction of acoustic radiation pressure from the input acoustic data over a portion of a surface enclosing all the acoustic sources, i.e., an open surface, becomes unique due to the unique continuation theory of elliptic equations. Hence the method is more stable and the reconstructed acoustic pressure is less dependable on the locations of the input acoustic data. Furthermore, the proposed method can be applied for both inverse and forward problems up to the minimum sphere enclosing the sources of interest. The effectiveness of the method is demonstrated by the results of several acoustic radiation examples with single or multi-frequency source in a two-dimensional configuration. The results from the method also show a measurable improvement in accuracy and consistency of reconstructed acoustic radiation pressure, in particular when the effect of the signal-to-noise ratio is included. PMID- 18396999 TI - Nonlinear source-filter coupling in phonation: vocal exercises. AB - Nonlinear source-filter coupling has been demonstrated in computer simulations, in excised larynx experiments, and in physical models, but not in a consistent and unequivocal way in natural human phonations. Eighteen subjects (nine adult males and nine adult females) performed three vocal exercises that represented a combination of various fundamental frequency and formant glides. The goal of this study was to pinpoint the proportion of source instabilities that are due to nonlinear source-tract coupling. It was hypothesized that vocal fold vibration is maximally destabilized when F(0) crosses F(1), where the acoustic load changes dramatically. A companion paper provides the theoretical underpinnings. Expected manifestations of a source-filter interaction were sudden frequency jumps, subharmonic generation, or chaotic vocal fold vibrations that coincide with F(0) F(1) crossovers. Results indicated that the bifurcations occur more often in phonations with F(0)-F(1) crossovers, suggesting that nonlinear source-filter coupling is partly responsible for source instabilities. Furthermore it was observed that male subjects show more bifurcations in phonations with F(0)-F(1) crossovers, presumably because in normal speech they are less likely to encounter these crossovers as much as females and hence have less practice in suppressing unwanted instabilities. PMID- 18397000 TI - Optimized translation of microbubbles driven by acoustic fields. AB - The problem of a single acoustically driven bubble translating unsteadily in a fluid is considered. The investigation of the translation equation identifies the inverse Reynolds number as a small perturbation parameter. The objective is to obtain a closed-form, leading order solution for the translation of the bubble, assuming nonlinear radial oscillations and a pressure field as the forcing term. In a second part, the periodic attractor of the Rayleigh-Plesset equation serves as basis for an optimal acoustic forcing designed to achieve maximized bubble translation over one dimensionless period. At near-resonant or super-resonant driving frequencies, it seems one cannot improve much on sinusoidal forcing. However at moderate acoustic intensity and sub-resonant frequencies, acoustic wave forms that enhance bubble collapse lead to displacement many times larger than the case of purely sinusoidal forcing. The survey covers a wide spectrum of driving ratios and bubble diameters including those relevant to biomedical applications. Shape stability issues are considered. Together, these results suggest new ways to predict some of the direct and indirect effects of the acoustic radiation force in applications such as targeted drug delivery, selective bubble driving, and accumulation. PMID- 18397001 TI - Sound velocity and attenuation in bubbly gels measured by transmission experiments. AB - Measurements of the phase velocity and attenuation of sound in concentrated samples of bubbly gels are presented. Hair gel was used as a matrix material to obtain well characterized distributions of bubbles. Ultrasonic measurements were conducted over a large range of frequencies, including the resonance frequencies of the bubbles. Surprisingly good agreement with Foldy's prediction was found, even for monodisperse samples at resonance frequencies, up to volume fraction of 1%. Beyond this concentration, the effects of high-order multiple scattering were observed. These results support the feasability of ultrasonic techniques to investigate the size distribution of bubbles in a weak gel or liquid. PMID- 18397002 TI - Acoustics of marine sediment under compaction: binary grain-size model and viscoelastic extension of Biot's theory. AB - This paper presents a model of acoustic wave propagation in unconsolidated marine sediment, including compaction, using a concept of a simplified sediment structure, modeled as a binary grain-size sphere pack. Compressional- and shear wave velocities and attenuation follow from a combination of Biot's model, used as the general framework, and two viscoelastic extensions resulting in complex grain and frame moduli, respectively. An effective-grain model accounts for the viscoelasticity arising from local fluid flow in expandable clay minerals in clay bearing sediments. A viscoelastic-contact model describes local fluid flow at the grain contacts. Porosity, density, and the structural Biot parameters (permeability, pore size, structure factor) as a function of pressure follow from the binary model, so that the remaining input parameters to the acoustic model consist solely of the mass fractions and the known mechanical properties of each constituent (e.g., carbonates, sand, clay, and expandable clay) of the sediment, effective pressure, or depth, and the environmental parameters (water depth, salinity, temperature). Velocity and attenuation as a function of pressure from the model are in good agreement with data on coarse- and fine-grained unconsolidated marine sediments. PMID- 18397003 TI - Spatial averaging of oceanic rainfall variability using underwater sound: Ionian Sea rainfall experiment 2004. AB - An experiment to evaluate the inherent spatial averaging of the underwater acoustic signal from rainfall was conducted in the winter of 2004 in the Ionian Sea southwest of Greece. A mooring with four passive aquatic listeners (PALs) at 60, 200, 1000, and 2000 m was deployed at 36.85 degrees N, 21.52 degrees E, 17 km west of a dual-polarization X-band coastal radar at Methoni, Greece. The acoustic signal is classified into wind, rain, shipping, and whale categories. It is similar at all depths and rainfall is detected at all depths. A signal that is consistent with the clicking of deep-diving beaked whales is present 2% of the time, although there was no visual confirmation of whale presence. Co-detection of rainfall with the radar verifies that the acoustic detection of rainfall is excellent. Once detection is made, the correlation between acoustic and radar rainfall rates is high. Spatial averaging of the radar rainfall rates in concentric circles over the mooring verifies the larger inherent spatial averaging of the rainfall signal with recording depth. For the PAL at 2000 m, the maximum correlation was at 3-4 km, suggesting a listening area for the acoustic rainfall measurement of roughly 30-50 km(2). PMID- 18397004 TI - Finite element simulation of the generation and detection by air-coupled transducers of guided waves in viscoelastic and anisotropic materials. AB - The measured characteristics (efficiency and sensitivity) of two air-coupled transducers allow for the prediction of the absolute values of the pressure of the bulk waves generated in air and for the measurement of the pressure of the field radiated in air by guided waves propagating in a structure. With finite element software, the pressure field generated by an air-coupled transducer is simulated by introducing a right-hand side member in the Helmholtz equation, which is used for computing the propagation from the transducer to a plate. The simulated source is rotated in order to impose an angle of incidence with respect to the normal of the plate and generate the corresponding guided mode. Inside the plate, the propagation is simulated with the dynamic equations of equilibrium and a complex stiffness tensor to take into account the viscoelastic anisotropy of the material. For modeling the three-dimensional fields of the guided modes propagating in a two-dimensional non-symmetry plane, a 2.5 dimensional model is introduced. The model computes the value of the pressure field radiated in air by the plates for any guided modes and can predict the detectability of the system for a known defect in a structure. A test bed incorporating two air-coupled transducers is used to generate and receive various guided modes. Two plates made of Perspex and carbon-epoxy composite are tested. The pressure measured by the receiver at various positions is compared to the results of the model to validate it. PMID- 18397005 TI - Piezoacoustic wave spectra using improved surface impedance matrix: application to high impedance-contrast layered plates. AB - Starting from the general modal solutions for a homogeneous layer of arbitrary material and crystalline symmetry, a matrix formalism is developed to establish the semianalytical expressions of the surface impedance matrices (SIM) for a single piezoelectric layer. By applying the electrical boundary conditions, the layer impedance matrix is reduced to a unified elastic form whether the material is piezoelectric or not. The characteristic equation for the dispersion curves is derived in both forms of a three-dimensional acoustic SIM and of an electrical scalar function. The same approach is extended to multilayered structures such as a piezoelectric layer sandwiched in between two metallic electrodes, a Bragg coupler, and a semi-infinite substrate as well. The effectiveness of the approach is numerically demonstrated by its ability to determine the full spectra of guided modes, even at extremely high frequencies, in layered plates comprising up to four layers and three materials. Negative slope in f-k curve for some modes, asymptotic behavior at short wavelength regime, as well as wave confinement phenomena made evident by the numerical results are analyzed and interpreted in terms of the surface acoustic waves and of the interfacial waves in connection with the bulk waves in massive materials. PMID- 18397006 TI - Acoustic energy harvesting using an electromechanical Helmholtz resonator. AB - This paper presents the development of an acoustic energy harvester using an electromechanical Helmholtz resonator (EMHR). The EMHR consists of an orifice, cavity, and a piezoelectric diaphragm. Acoustic energy is converted to mechanical energy when sound incident on the orifice generates an oscillatory pressure in the cavity, which in turns causes the vibration of the diaphragm. The conversion of acoustic energy to electrical energy is achieved via piezoelectric transduction in the diaphragm of the EMHR. Moreover, the diaphragm is coupled with energy reclamation circuitry to increase the efficiency of the energy conversion. Lumped element modeling of the EMHR is used to provide physical insight into the coupled energy domain dynamics governing the energy reclamation process. The feasibility of acoustic energy reclamation using an EMHR is demonstrated in a plane wave tube for two power converter topologies. The first is comprised of only a rectifier, and the second uses a rectifier connected to a flyback converter to improve load matching. Experimental results indicate that approximately 30 mW of output power is harvested for an incident sound pressure level of 160 dB with a flyback converter. Such power level is sufficient to power a variety of low power electronic devices. PMID- 18397007 TI - Adaptive wave field synthesis for active sound field reproduction: experimental results. AB - Sound field reproduction has applications in music reproduction, spatial audio, sound environment reproduction, and experimental acoustics. Sound field reproduction can be used to artificially reproduce the spatial character of natural hearing. The objective is then to reproduce a sound field in a real reproduction environment. Wave field synthesis (WFS) is a known open-loop technology which assumes that the reproduction environment is anechoic. The room response thus reduces the quality of the physical sound field reproduction by WFS. In recent research papers, adaptive wave field synthesis (AWFS) was defined as a potential solution to compensate for these quality reductions from which WFS objective performance suffers. In this paper, AWFS is experimentally investigated as an active sound field reproduction system with a limited number of reproduction error sensors to compensate for the response of the listening environment. Two digital signal processing algorithms for AWFS are used for comparison purposes, one of which is based on independent radiation mode control. AWFS performed propagating sound field reproduction better than WFS in three tested reproduction spaces (hemianechoic chamber, standard laboratory space, and reverberation chamber). PMID- 18397008 TI - Adaptive wave field synthesis for broadband active sound field reproduction: signal processing. AB - Sound field reproduction is a physical approach to the reproduction of the natural spatial character of hearing. It is also useful in experimental acoustics and psychoacoustics. Wave field synthesis (WFS) is a known open-loop technology which assumes that the reproduction environment is anechoic. A real reflective reproduction space thus reduces the objective accuracy of WFS. Recently, adaptive wave field synthesis (AWFS) was defined as a combination of WFS and active compensation. AWFS is based on the minimization of reproduction errors and on the penalization of departure from the WFS solution. This paper focuses on signal processing for AWFS. A classical adaptive algorithm is modified for AWFS: filtered-reference least-mean-square. This modified algorithm and the classical equivalent leaky algorithm have similar convergence properties except that the WFS solution influences the adaptation rule of the modified algorithm. The paper also introduces signal processing for independent radiation mode control of AWFS on the basis of plant decoupling. Simulation results for AWFS are introduced for free-field and reflective spaces. The two algorithms effectively reproduce the sound field and compensate for the reproduction errors at the error sensors. The independent radiation mode control allows a more flexible tuning of the algorithm. PMID- 18397009 TI - Effect of damping on the propensity of squeal instability: an experimental investigation. AB - Friction induced vibrations in automotive brakes is recognized as a major problem in industry. Squeal is a difficult subject because of its unpredictability caused by a not completely understood sensitivity to variation of the system parameters. In the literature several analytical and numerical studies deal with the relationship between damping and system propensity to have instability. These studies highlight the existence of a nonintuitive effect of damping distribution on modal coupling that gives rise to the unstable vibrations. The complexity of commercial brakes and the difficulties to identify the values of modal damping in brake assemblies lead to the necessity to rely on experimental analysis using simplified test rigs. This paper presents an experimental investigation of the relationship between the distribution of modal damping and the propensity to develop squeal in a beam-on-disk setup, which reliably reproduces squeal events with easy control and measurement of the damping of the disk and the beam, respectively. The experiments highlight the key role played by the modal damping distribution on squeal: A nonuniform repartition of the modal damping causes an increase of the squeal propensity. PMID- 18397010 TI - A model for sonar interrogation of complex bottom and surface targets in shallow water waveguides. AB - Many problems of current interest in underwater acoustics involve low-frequency broadband sonar interrogation of objects near the sea surface or sea floor of a shallow-water environment. When the target is situated near the upper or lower boundary of the water column the acoustic interactions with the target objects are complicated by interactions with the nearby free surface or fluid-sediment interface, respectively. A practical numerical method to address such situations is presented. The model provides high levels of accuracy with the flexibility to handle complex, three-dimensional targets in range-independent environments. The model is demonstrated using several bottom target scenarios, with and without locally undulating seabeds. The impact of interface and boundary interactions is considered with an eye toward using the sonar return signal as the basis for acoustic imaging or spectral classification. PMID- 18397011 TI - Luke and power residue sequence diffusers. AB - Conventional Schroeder diffusers have been successfully used for many years. However, their frequency range is limited by the flat plate effect that occurs when all the wells radiate in phase. This occurs at harmonics of p times the design frequency f(0), where p is the small prime that is used to generate the structure. A typical diffuser, using p=7 and f(0)=500 Hz, has an upper frequency limit of only 3.5 kHz. Achieving a first flat plate frequency above 20 kHz requires a prime equal to at least 41 and results in diffusers that are too big to be practical in most applications. This paper suggests an alternative approach using number theoretic sequences that, although short in length, are based on large integers. Two new sequences, Type-II Luke and power residue, have this desired characteristic. They are investigated using both simple models and the more exact boundary element method. The results show the flat plate effect is moved to much higher frequencies as expected. For Luke sequences at certain frequencies, redirection rather than dispersion is achieved. Modulation techniques can be used to mitigate these problems. Power residue sequences perform the best, providing good diffusion and a flat plate frequency outside the audible range. PMID- 18397012 TI - Acoustic model for robustness analysis of optimal multipoint room equalization. AB - In this paper, an acoustic model for the robustness analysis of optimal multipoint room equalization is proposed. The optimal multipoint equalization aims to have the optimal performance in a least-squares sense for all measured points. The model can be used for theoretical robustness estimation depending on the critical design parameters such as the number of measurement points, the distance between measurements, or the frequency before applying real equalization system. The analysis results show that it is important to set the appropriate number of measurement points and the distances between measurement points to ensure the enlarged equalization region at a specific frequency. PMID- 18397013 TI - Expressions of dissipated powers and stored energies in poroelastic media modeled by {u,U} and {u,P} formulations. AB - This paper is devoted to the rigorous obtention of the energy balance in porous materials. The wave propagation in the porous media is described by Biot-Allard's {u,U} and {u,P} formulations. The paper derives the expressions for stored kinetic and strain energies together with dissipated energies. It is shown that, in the case of mixed formulations, these expressions do not correspond to the real and imaginary parts of the variational formulations. A quantitative convergence analysis of finite element scheme is then undertaken with the help of these indicators. It is shown that the order of convergence of these indicators for linear finite-element is one and that they are then well fitted to check the validity of finite-element models. PMID- 18397014 TI - Evaluation of acoustical conditions for speech communication in working elementary school classrooms. AB - Detailed acoustical measurements were made in 41 working elementary school classrooms near Ottawa, Canada to obtain more representative and more accurate indications of the acoustical quality of conditions for speech communication during actual teaching activities. This paper describes the room acoustics characteristics and noise environment of 27 traditional rectangular classrooms from the 41 measured rooms. The purpose of the work was to better understand how to improve speech communication between teachers and students. The study found, that on average, the students experienced: teacher speech levels of 60.4 dB A, noise levels of 49.1 dB A, and a mean speech-to-noise ratio of 11 dB A during teaching activities. The mean reverberation time in the occupied classrooms was 0.41 s, which was 10% less than in the unoccupied rooms. The reverberation time measurements were used to determine the average absorption added by each student. Detailed analyses of early and late-arriving speech sounds showed these sound levels could be predicted quite accurately and suggest improved approaches to room acoustics design. PMID- 18397015 TI - The intelligibility of speech in elementary school classrooms. AB - This is the second of two papers describing the results of acoustical measurements and speech intelligibility tests in elementary school classrooms. The intelligibility tests were performed in 41 classrooms in 12 different schools evenly divided among grades 1, 3, and 6 students (nominally 6, 8, and 11 year olds). Speech intelligibility tests were carried out on classes of students seated at their own desks in their regular classrooms. Mean intelligibility scores were significantly related to signal-to-noise ratios and to the grade of the students. While the results are different than those from some previous laboratory studies that included less realistic conditions, they agree with previous in-classroom experiments. The results indicate that +15 dB signal-to noise ratio is not adequate for the youngest children. By combining the speech intelligibility test results with measurements of speech and noise levels during actual teaching situations, estimates of the fraction of students experiencing near-ideal acoustical conditions were made. The results are used as a basis for estimating ideal acoustical criteria for elementary school classrooms. PMID- 18397016 TI - Relationship between listening difficulty and acoustical objective measures in reverberant sound fields. AB - The previous work [Morimoto et al., J. Acoust. Soc. Am. 116, 1607-1613] showed that listening difficulty ratings can be used to evaluate speech transmission performance more exactly and sensitively than intelligibility. Meanwhile, speech transmission performance is usually evaluated using acoustical objective measures, which are directly associated with physical parameters of room acoustic design. However, the relationship between listening difficulty ratings and acoustical objective measures was not minutely investigated. In the present study, a total of 96 impulse responses were used to investigate the relationship between listening difficulty ratings and several objective measures in unidirectional sound fields. The result of the listening test showed that (1) the correlation between listening difficulty ratings and speech transmission index (STI) is the strongest of all tested objective measures, and (2) A-weighted D(50), C(50), and center time, which are obtained from the impulse responses passed through an A-weighted filter, also strongly correlate with listening difficulty ratings, and their correlations with listening difficulty ratings are not statistically different from the correlation between listening difficulty ratings and STI. PMID- 18397017 TI - Two-dimensional poroelastic acoustical foam shape design for absorption coefficient maximization by topology optimization method. AB - Optimal shape design of a two-dimensional poroelastic acoustical foam is formulated as a topology optimization problem. For a poroelastic acoustical system consisting of an air region and a poroelastic foam region, two different physical regions are continuously changed in an iterative design process. To automatically account for the moving interfaces between two regions, we propose a new unified model to analyze the whole poroelastic acoustical foam system with one set of governing equations; Biot's equations are modified with a material property interpolation from a topology optimization method. With the unified analysis model, we carry out two-dimensional optimal shape design of a poroelastic acoustical foam by a gradient-based topology optimization setting. The specific objective is the maximization of the absorption coefficient in low and middle ranges of frequencies with different amounts of a poroelastic material. The performances of the obtained shapes are compared with those of well known wedge shapes, and the improvement of absorption is physically interpreted. PMID- 18397018 TI - Causal impulse response for circular sources in viscous media. AB - The causal impulse response of the velocity potential for the Stokes wave equation is derived for calculations of transient velocity potential fields generated by circular pistons in viscous media. The causal Green's function is numerically verified using the material impulse response function approach. The causal, lossy impulse response for a baffled circular piston is then calculated within the near field and the far field regions using expressions previously derived for the fast near field method. Transient velocity potential fields in viscous media are computed with the causal, lossy impulse response and compared to results obtained with the lossless impulse response. The numerical error in the computed velocity potential field is quantitatively analyzed for a range of viscous relaxation times and piston radii. Results show that the largest errors are generated in locations near the piston face and for large relaxation times, and errors are relatively small otherwise. Unlike previous frequency-domain methods that require numerical inverse Fourier transforms for the evaluation of the lossy impulse response, the present approach calculates the lossy impulse response directly in the time domain. The results indicate that this causal impulse response is ideal for time-domain calculations that simultaneously account for diffraction and quadratic frequency-dependent attenuation in viscous media. PMID- 18397019 TI - An acoustical array combining microphones and piezoelectric devices. AB - This paper describes an acoustical array combining microphones and piezoelectric devices. Conventional microphone arrays have been widely utilized to realize noise reduction, sound separation and direction of arrival estimation system. However, when a conventional microphone array is mounted on a real system, such as a machine, vehicle or robot, the microphones are set extremely close to the system's actual body. In such cases, the noise from the system itself, such as motors, gears, and engines, namely internal noise, often becomes a troublesome problem. It is difficult to reduce internal noise utilizing a conventional microphone array because internal noise sources are extremely close to the microphones. As internal noise is not always stationary, statistically independent or sparse, most useful blind source separation approaches, such as independent component analysis and the sparseness approach, cannot be employed. Our aim is to reduce internal noise utilizing microphones and piezoelectric devices attached to the internal noise source. In this paper, a general description of the acoustical array is formulated and the characteristic features of microphones and piezoelectric devices in an acoustical array are given. An acoustical array combining microphones and piezoelectric devices is also described with some experimental results. PMID- 18397020 TI - Bending wavelet for flexural impulse response. AB - The work addresses the definition of a wavelet that is adapted to analyze a flexural impulse response of a beam or plate that can be modeled with the Euler Bernoulli bending theory. The wavelet gives the opportunity to directly analyze the dispersion characteristics of a pulse. The aim is to localize a source or to measure material parameters. An overview of the mathematical properties of the wavelet is presented. An algorithm for the optimal extraction of the dispersion characteristics with the use of genetic algorithms is outlined. The application of the wavelet is shown in an example and experiment. PMID- 18397021 TI - Localization of multiple acoustic sources with small arrays using a coherence test. AB - Direction finding of more sources than sensors is appealing in situations with small sensor arrays. Potential applications include surveillance, teleconferencing, and auditory scene analysis for hearing aids. A new technique for time-frequency-sparse sources, such as speech and vehicle sounds, uses a coherence test to identify low-rank time-frequency bins. These low-rank bins are processed in one of two ways: (1) narrowband spatial spectrum estimation at each bin followed by summation of directional spectra across time and frequency or (2) clustering low-rank covariance matrices, averaging covariance matrices within clusters, and narrowband spatial spectrum estimation of each cluster. Experimental results with omnidirectional microphones and colocated directional microphones demonstrate the algorithm's ability to localize 3-5 simultaneous speech sources over 4 s with 2-3 microphones to less than 1 degree of error, and the ability to localize simultaneously two moving military vehicles and small arms gunfire. PMID- 18397022 TI - Angle-dependent ultrasonic detection and imaging of brachytherapy seeds using singular spectrum analysis. AB - Transrectal-ultrasound-guided brachytherapy uses small titanium-shelled radioactive seeds to locally treat prostate cancer. During the implantation procedure, needles inserted transperitoneally cause gland movement resulting in seed misplacement and suboptimal dosimetry. In a previous study, an algorithm based on singular spectrum analysis (SSA) applied to envelope-detected ultrasound signals was proposed to determine seed locations [J. Mamou and E. J. Feleppa, J. Acoust. Soc. Am. 121, 1790-1801 (2007)]. Successful implementation of the SSA algorithm could allow correcting dosimetry errors during the implantation procedure. The algorithm demonstrated promise when the seed orientation was parallel to the needle and normal to the ultrasound beam. In this present study, the algorithm was tested when the seed orientation deviated up to 22 degrees from normality. Experimental data from a seed in an ideal environment and in beef were collected with a single-element, spherically focused, 5 MHz transducer. Simulations were designed and evaluated with the algorithm. Finally, objective quantitative scoring metrics were developed to evaluate the algorithm performance and for comparison with B-mode images. The results quantitatively established that the SSA algorithm always outperformed B-mode images and that seeds could be detected accurately up to a deviation of approximately 10 degrees . PMID- 18397023 TI - Orthotropic material properties of the gerbil basilar membrane. AB - In this paper, two sets of experimental results to extract the two effective elastic moduli, the effective shear modulus, and the effective Poisson's ratio for the gerbil cochlear partition are analyzed. In order to accomplish this, a geometrically nonlinear composite orthotropic plate model is employed. The model is used to predict both out-of-plane and in-plane motion of the partition under a static finite area distributed load. This loading condition models the small, but finite size, probe tips used in experiments. Both in-plane and out-of-plane motion are needed for comparison with recent experimental results. It is shown that the spatial decay rate (the space constant) for the in-plane deflection is different than for the out-of-plane deflection, which has a significant effect on the derived partition properties. The size of the probe tip is shown to have little influence on the results. Results are presented for two types of boundary conditions. Orthotropy ratios determined from the experimental data are found to vary with longitudinal position and choice of boundary conditions. Orthotropy ratios (the ratio of the two elastic moduli) are in the range of 65 close to the base to 10 in the upper middle turn of the cochlea. PMID- 18397025 TI - Cortical sensitivity to periodicity of speech sounds. AB - Previous non-invasive brain research has reported auditory cortical sensitivity to periodicity as reflected by larger and more anterior responses to periodic than to aperiodic vowels. The current study investigated whether there is a lower fundamental frequency (F0) limit for this effect. Auditory evoked fields (AEFs) elicited by natural-sounding 400 ms periodic and aperiodic vowel stimuli were measured with magnetoencephalography. Vowel F0 ranged from normal male speech (113 Hz) to exceptionally low values (9 Hz). Both the auditory N1m and sustained fields were larger in amplitude for periodic than for aperiodic vowels. The AEF sources for periodic vowels were also anterior to those for the aperiodic vowels. Importantly, the AEF amplitudes and locations were unaffected by the F0 decrement of the periodic vowels. However, the N1m latency increased monotonically as F0 was decreased down to 19 Hz, below which this trend broke down. Also, a cascade of transient N1m-like responses was observed in the lowest F0 condition. Thus, the auditory system seems capable of extracting the periodicity even from very low F0 vowels. The behavior of the N1m latency and the emergence of a response cascade at very low F0 values may reflect the lower limit of pitch perception. PMID- 18397024 TI - Low-frequency and high-frequency distortion product otoacoustic emission suppression in humans. AB - Distortion product otoacoustic emission suppression (quantified as decrements) was measured for f(2)=500 and 4000 Hz, for a range of primary levels (L(2)), suppressor frequencies (f(3)), and suppressor levels (L(3)) in 19 normal-hearing subjects. Slopes of decrement-versus-L(3) functions were similar at both f(2) frequencies, and decreased as f(3) increased. Suppression tuning curves, constructed from decrement functions, were used to estimate (1) suppression for on- and low-frequency suppressors, (2) tip-to-tail differences, (3) Q(ERB), and (4) best frequency. Compression, estimated from the slope of functions relating suppression "threshold" to L(2) for off-frequency suppressors, was similar for 500 and 4000 Hz. Tip-to-tail differences, Q(ERB), and best frequency decreased as L(2) increased for both frequencies. However, tip-to-tail difference (an estimate of cochlear-amplifier gain) was 20 dB greater at 4000 Hz, compared to 500 Hz. Q(ERB) decreased to a greater extent with L(2) when f(2)=4000 Hz, but, on an octave scale, best frequency shifted more with level when f(2)=500 Hz. These data indicate that, at both frequencies, cochlear processing is nonlinear. Response growth and compression are similar at the two frequencies, but gain is greater at 4000 Hz and spread of excitation is greater at 500 Hz. PMID- 18397026 TI - Temporary hearing loss influences post-stimulus time histogram and single neuron action potential estimates from human compound action potentials. AB - An analytic compound action potential (CAP) obtained by convolving functional representations of the post-stimulus time histogram summed across auditory nerve neurons [P(t)] and a single neuron action potential [U(t)] was fit to human CAPs. The analytic CAP fit to pre- and postnoise-induced temporary hearing threshold shift (TTS) estimated in vivo P(t) and U(t) and the number of neurons contributing to the CAPs (N). The width of P(t) decreased with increasing signal level and was wider at the lowest signal level following noise exposure. P(t) latency decreased with increasing signal level and was shorter at all signal levels following noise exposure. The damping and oscillatory frequency of U(t) increased with signal level. For subjects with large amounts of TTS, U(t) had greater damping than before noise exposure particularly at low signal levels. Additionally, U(t) oscillation was lower in frequency at all click intensities following noise exposure. N increased with signal level and was smaller after noise exposure at the lowest signal level. Collectively these findings indicate that neurons contributing to the CAP during TTS are fewer in number, shorter in latency, and poorer in synchrony than before noise exposure. Moreover, estimates of single neuron action potentials may decay more rapidly and have a lower oscillatory frequency during TTS. PMID- 18397027 TI - Comodulation detection differences in children and adults. AB - This study investigated comodulation detection differences (CDD) in children (ages 4.8-10.1 years) and adults. The signal was 30-Hz wide band of noise centered on 2 kHz, and the masker consisted of six 30-Hz wide bands of noise spanning center frequencies from 870 to 4160 Hz. The envelopes of the masking bands were always comodulated, and the envelope of the signal was either comodulated or random with respect to the masker. In some conditions, the maskers were gated on prior to the signal in order to minimize effects related to perceptual fusion of the signal and masker. CDD was computed as the difference between signal detection thresholds in conditions where all bands were comodulated and conditions where the envelope of the signal was random with respect to the envelopes of the maskers. Values of CDD were generally small in children compared to adults. In contrast, masking release related to masker/signal onset asynchrony was comparable across age groups. The small CDDs in children are discussed in terms of sensitivity to comodulation as a perceptual fusion cue and informational masking associated with the detection of a signal in a complex background, an effect that is ameliorated by asynchronous onset. PMID- 18397028 TI - Perception of suprathreshold amplitude modulation and intensity increments: Weber's law revisited. AB - The perceived strength of intensity fluctuations evoked by suprathreshold sinusoidal amplitude modulation (AM) and the perceived size of intensity increments were compared across levels of a wideband noise and a 1-kHz tone. For the 1-kHz tone, the comparisons were made in quiet and in a high-pass noise. The data indicate that suprathreshold modulation depths and intensity increments, perceived as equivalent across levels, follow a pattern resembling Weber's law for noise and the "near miss" to Weber's law for a tone. The effect of a high pass noise was largely consistent with that observed for AM and increment detection. The data suggest that Weber's law is not a direct consequence of the dependence of internal noise on stimulus level, as suggested by multiplicative internal noise models. Equal loudness ratios and equal loudness differences (computed using loudness for the stationary portions before and after the increment) accounted for the increment-matching data for noise and for the tone, respectively, but neither measure predicted the results for both types of stimuli. Predictions based on log-transformed excitation patterns and predictions using an equal number of intensity just-noticeable differences were in qualitative, but not quantitative, agreement with the data. PMID- 18397029 TI - Speech segregation in rooms: monaural, binaural, and interacting effects of reverberation on target and interferer. AB - Speech reception thresholds were measured in virtual rooms to investigate the influence of reverberation on speech intelligibility for spatially separated targets and interferers. The measurements were realized under headphones, using target sentences and noise or two-voice interferers. The room simulation allowed variation of the absorption coefficient of the room surfaces independently for target and interferer. The direct-to-reverberant ratio and interaural coherence of sources were also varied independently by considering binaural and diotic listening. The main effect of reverberation on the interferer was binaural and mediated by the coherence, in agreement with binaural unmasking theories. It appeared at lower reverberation levels than the effect of reverberation on the target, which was mainly monaural and associated with the direct-to-reverberant ratio, and could be explained by the loss of amplitude modulation in the reverberant speech signals. This effect was slightly smaller when listening binaurally. Reverberation might also be responsible for a disruption of the mechanism by which the auditory system exploits fundamental frequency differences to segregate competing voices, and a disruption of the "listening in the gaps" associated with speech interferers. These disruptions may explain an interaction observed between the effects of reverberation on the targets and two-voice interferers. PMID- 18397030 TI - Binaural speech unmasking and localization in noise with bilateral cochlear implants using envelope and fine-timing based strategies. AB - Four adult bilateral cochlear implant users, with good open-set sentence recognition, were tested with three different sound coding strategies for binaural speech unmasking and their ability to localize 100 and 500 Hz click trains in noise. Two of the strategies tested were envelope-based strategies that are clinically widely used. The third was a research strategy that additionally preserved fine-timing cues at low frequencies. Speech reception thresholds were determined in diotic noise for diotic and interaurally time-delayed speech using direct audio input to a bilateral research processor. Localization in noise was assessed in the free field. Overall results, for both speech and localization tests, were similar with all three strategies. None provided a binaural speech unmasking advantage due to the application of 700 micros interaural time delay to the speech signal, and localization results showed similar response patterns across strategies that were well accounted for by the use of broadband interaural level cues. The data from both experiments combined indicate that, in contrast to normal hearing, timing cues available from natural head-width delays do not offer binaural advantages with present methods of electrical stimulation, even when fine-timing cues are explicitly coded. PMID- 18397031 TI - Effects of in-the-ear microphone directionality on sound direction identification. AB - As advanced signal processing algorithms have been proposed to enhance hearing protective device (HPD) performance, it is important to determine how directional microphones might affect the localization ability of users and whether they might cause safety hazards. The effect of in-the-ear microphone directivity was assessed by measuring sound source identification of speech in the horizontal plane. Recordings of speech in quiet and in noise were made with Knowles Electronic Manikin for Acoustic Research wearing bilateral in-the-ear hearing aids with microphones having adjustable directivity (omnidirectional, cardioid, hypercardioid, supercardioid). Signals were generated from 16 locations in a circular array. Sound direction identification performance of eight normal hearing listeners and eight hearing-impaired listeners revealed that directional microphones did not degrade localization performance and actually reduced the front-back and lateral localization errors made when listening through omnidirectional microphones. The summed rms speech level for the signals entering the two ears appear to serve as a cue for making front-back discriminations when using directional microphones in the experimental setting. The results of this study show that the use of matched directional microphones when worn bilaterally do not have a negative effect on the ability to localize speech in the horizontal plane and may thus be useful in HPD design. PMID- 18397032 TI - Pulse-rate discrimination by cochlear-implant and normal-hearing listeners with and without binaural cues. AB - Experiment 1 measured rate discrimination of electric pulse trains by bilateral cochlear implant (CI) users, for standard rates of 100, 200, and 300 pps. In the diotic condition the pulses were presented simultaneously to the two ears. Consistent with previous results with unilateral stimulation, performance deteriorated at higher standard rates. In the signal interval of each trial in the dichotic condition, the standard rate was presented to the left ear and the (higher) signal rate was presented to the right ear; the non-signal intervals were the same as in the diotic condition. Performance in the dichotic condition was better for some listeners than in the diotic condition for standard rates of 100 and 200 pps, but not at 300 pps. It is concluded that the deterioration in rate discrimination observed for CI users at high rates cannot be alleviated by the introduction of a binaural cue, and is unlikely to be limited solely by central pitch processes. Experiment 2 performed an analogous experiment in which 300-pps acoustic pulse trains were bandpass filtered (3900-5400 Hz) and presented in a noise background to normal-hearing listeners. Unlike the results of experiment 1, performance was superior in the dichotic than in the diotic condition. PMID- 18397033 TI - A glimpsing account for the benefit of simulated combined acoustic and electric hearing. AB - The benefits of combined electric and acoustic stimulation (EAS) in terms of speech recognition in noise are well established; however the underlying factors responsible for this benefit are not clear. The present study tests the hypothesis that having access to acoustic information in the low frequencies makes it easier for listeners to glimpse the target. Normal-hearing listeners were presented with vocoded speech alone (V), low-pass (LP) filtered speech alone, combined vocoded and LP speech (LP+V) and with vocoded stimuli constructed so that the low-frequency envelopes were easier to glimpse. Target speech was mixed with two types of maskers (steady-state noise and competing talker) at -5 to 5 dB signal-to-noise ratios. Results indicated no advantage of LP+V in steady noise, but a significant advantage over V in the competing talker background, an outcome consistent with the notion that it is easier for listeners to glimpse the target in fluctuating maskers. A significant improvement in performance was noted with the modified glimpsed stimuli over the original vocoded stimuli. These findings taken together suggest that a significant factor contributing to the EAS advantage is the enhanced ability to glimpse the target. PMID- 18397035 TI - Incorporation of phonetic constraints in acoustic-to-articulatory inversion. AB - This study investigates the use of constraints upon articulatory parameters in the context of acoustic-to-articulatory inversion. These speaker independent constraints, referred to as phonetic constraints, were derived from standard phonetic knowledge for French vowels and express authorized domains for one or several articulatory parameters. They were experimented on in an existing inversion framework that utilizes Maeda's articulatory model and a hypercubic articulatory-acoustic table. Phonetic constraints give rise to a phonetic score rendering the phonetic consistency of vocal tract shapes recovered by inversion. Inversion has been applied to vowels articulated by a speaker whose corresponding x-ray images are also available. Constraints were evaluated by measuring the distance between vocal tract shapes recovered through inversion to real vocal tract shapes obtained from x-ray images, by investigating the spreading of inverse solutions in terms of place of articulation and constriction degree, and finally by studying the articulatory variability. Results show that these constraints capture interdependencies and synergies between speech articulators and favor vocal tract shapes close to those realized by the human speaker. In addition, this study also provides how acoustic-to-articulatory inversion can be used to explore acoustical and compensatory articulatory properties of an articulatory model. PMID- 18397034 TI - Effects of upper-frequency boundary and spectral warping on speech intelligibility in electrical stimulation. AB - Speech understanding was tested for seven listeners using 12-electrode Med-El cochlear implants (CIs) and six normal-hearing listeners using a CI simulation. Eighteen different types of processing were evaluated, which varied the frequency to-tonotopic place mapping and the upper boundary of the frequency and stimulation range. Spectrally unwarped and warped conditions were included. Unlike previous studies on this topic, the lower boundary of the frequency and stimulation range was fixed while the upper boundary was varied. For the unwarped conditions, only eight to ten channels were needed in both quiet and noise to achieve no significant degradation in speech understanding compared to the normal 12-electrode speech processing. The unwarped conditions were often the best conditions for understanding speech; however, small changes in frequency-to-place mapping (<0.77 octaves for the most basal electrode) yielded no significant degradation in performance from the nearest unwarped condition. A second experiment measured the effect of feedback training for both the unwarped and warped conditions. Improvements were found for the unwarped and frequency expanded conditions, but not for the compressed condition. These results have implications for new CI processing strategies, such as the inclusion of spectral localization cues. PMID- 18397036 TI - Spatiotemporal classification of vocal fold dynamics by a multimass model comprising time-dependent parameters. AB - A model-based approach is proposed to objectively measure and classify vocal fold vibrations by left-right asymmetries along the anterior-posterior direction, especially in the case of nonstationary phonation. For this purpose, vocal fold dynamics are recorded in real time with a digital high-speed camera during phonation of sustained vowels as well as pitch raises. The dynamics of a multimass model with time-dependent parameters are matched to vocal fold vibrations extracted at dorsal, medial, and ventral positions by an automatic optimization procedure. The block-based optimization accounts for nonstationary vibrations and compares the vocal fold and model dynamics by wavelet coefficients. The optimization is verified with synthetically generated data sets and is applied to 40 clinical high-speed recordings comprising normal and pathological voice subjects. The resulting model parameters allow an intuitive visual assessment of vocal fold instabilities within an asymmetry diagram and are applicable to an objective quantification of asymmetries. PMID- 18397037 TI - A three-dimensional articulatory model of the velum and nasopharyngeal wall based on MRI and CT data. AB - An original three-dimensional (3D) linear articulatory model of the velum and nasopharyngeal wall has been developed from magnetic resonance imaging (MRI) and computed tomography images of a French subject sustaining a set of 46 articulations, covering his articulatory repertoire. The velum and nasopharyngeal wall are represented by generic surface triangular meshes fitted to the 3D contours extracted from MRI for each articulation. Two degrees of freedom were uncovered by principal component analysis: first, VL accounts for 83% of the velum variance, corresponding to an oblique vertical movement seemingly related to the levator veli palatini muscle; second, VS explains another 6% of the velum variance, controlling a mostly horizontal movement possibly related to the sphincter action of the superior pharyngeal constrictor. The nasopharyngeal wall is also controlled by VL for 47% of its variance. Electromagnetic articulographic data recorded on the velum fitted these parameters exactly, and may serve to recover dynamic velum 3D shapes. The main oral and nasopharyngeal area functions controlled by the articulatory model, complemented by the area functions derived from the complex geometry of each nasal passage extracted from coronal MRIs, were fed to an acoustic model and gave promising results about the influence of velum movements on the spectral characteristics of nasals. PMID- 18397039 TI - Spectral weighting strategies for hearing-impaired listeners measured using a correlational method. AB - Spectral weighting strategies using a correlational method [R. A. Lutfi, J. Acoust. Soc. Am. 97, 1333-1334 (1995); V. M. Richards and S. Zhu, J. Acoust. Soc. Am. 95, 423-424 (1994)] were measured in ten listeners with sensorineural-hearing loss on a sentence recognition task. Sentences and a spectrally matched noise were filtered into five separate adjacent spectral bands and presented to listeners at various signal-to-noise ratios (SNRs). Five point-biserial correlations were computed between the listeners' response (correct or incorrect) on the task and the SNR in each band. The stronger the correlation between performance and SNR, the greater that given band was weighted by the listener. Listeners were tested with and without hearing aids on. All listeners were experienced hearing aid users. Results indicated that the highest spectral band (approximately 2800-11 000 Hz) received the greatest weight in both listening conditions. However, the weight on the highest spectral band was less when listeners performed the task with their hearing aids on in comparison to when listening without hearing aids. No direct relationship was observed between the listeners' weights and the sensation level within a given band. PMID- 18397038 TI - Seeing pitch: visual information for lexical tones of Mandarin-Chinese. AB - Mandarin perceivers were tested in visual lexical-tone identification before and after learning. Baseline performance was only slightly above chance, although there appeared to be some visual information in the speakers' neck and head movements. When participants were taught to use this visible information in two experiments, visual tone identification improved significantly. There appears to be a relationship between the production of lexical tones and the visible movements of the neck, head, and mouth, and this information can be effectively used after a short training session. PMID- 18397040 TI - Using blind source separation techniques to improve speech recognition in bilateral cochlear implant patients. AB - Bilateral cochlear implants seek to restore the advantages of binaural hearing by improving access to binaural cues. Bilateral implant users are currently fitted with two processors, one in each ear, operating independent of one another. In this work, a different approach to bilateral processing is explored based on blind source separation (BSS) by utilizing two implants driven by a single processor. Sentences corrupted by interfering speech or speech-shaped noise are presented to bilateral cochlear implant users at 0 dB signal-to-noise ratio in order to evaluate the performance of the proposed BSS method. Subjects are tested in both anechoic and reverberant settings, wherein the target and masker signals are spatially separated. Results indicate substantial improvements in performance in both anechoic and reverberant settings over the subjects' daily strategies for both masker conditions and at various locations of the masker. It is speculated that such improvements are due to the fact that the proposed BSS algorithm capitalizes on the variations of interaural level differences and interaural time delays present in the mixtures of the signals received by the two microphones, and exploits that information to spatially separate the target from the masker signals. PMID- 18397041 TI - Measurement of vocal-tract influence during saxophone performance. AB - This paper presents experimental results that quantify the range of influence of vocal tract manipulations used in saxophone performance. The experiments utilized a measurement system that provides a relative comparison of the upstream windway and downstream air column impedances under normal playing conditions, allowing researchers and players to investigate the effect of vocal-tract manipulations in real time. Playing experiments explored vocal-tract influence over the full range of the saxophone, as well as when performing special effects such as pitch bending, multiphonics, and "bugling." The results show that, under certain conditions, players can create an upstream windway resonance that is strong enough to override the downstream system in controlling reed vibrations. This can occur when the downstream air column provides only weak support of a given note or effect, especially for notes with fundamental frequencies an octave below the air column cutoff frequency and higher. Vocal-tract influence is clearly demonstrated when pitch bending notes high in the traditional range of the alto saxophone and when playing in the saxophone's extended register. Subtle timbre variations via tongue position changes are possible for most notes in the saxophone's traditional range and can affect spectral content from at least 800 2000 Hz. PMID- 18397042 TI - Numerical simulation of a piano soundboard under downbearing. AB - A finite element model of a piano soundboard is used to study the effect of the strings tension (downbearing) on its vibration, considering the ribs, the bridges and the crown. The downbearing is modeled with the prestress theory. Prestress calculation with linear and nonlinear models including geometric rigidity are compared in terms of the modal frequencies. The effects of the downbearing in modal frequencies and mobility are investigated and the importance of the crown on these results is evaluated. A simple phenomenological law is exhibited, which characterizes the evolution of eigenfrequencies with downbearing, including the initial crown. PMID- 18397043 TI - Negative dispersion in bone: the role of interference in measurements of the apparent phase velocity of two temporally overlapping signals. AB - In this study the attenuation coefficient and dispersion (frequency dependence of phase velocity) are measured using a phase sensitive (piezoelectric) receiver in a phantom in which two temporally overlapping signals are detected, analogous to the fast and slow waves typically found in measurements of cancellous bone. The phantom consisted of a flat and parallel Plexiglas plate into which a step discontinuity was milled. The phase velocity and attenuation coefficient of the plate were measured using both broadband and narrowband data and were calculated using standard magnitude and phase spectroscopy techniques. The observed frequency dependence of the phase velocity and attenuation coefficient exhibit significant changes in their frequency dependences as the interrogating ultrasonic field is translated across the step discontinuity of the plate. Negative dispersion is observed at specific spatial locations of the plate at which the attenuation coefficient rises linearly with frequency, a behavior analogous to that of bone measurements reported in the literature. For all sites investigated, broadband and narrowband data (3-7 MHz) demonstrate excellent consistency. Evidence suggests that the interference between the two signals simultaneously reaching the phase sensitive piezoelectric receiver is responsible for this negative dispersion. PMID- 18397044 TI - Application of Biot's theory to ultrasonic characterization of human cancellous bones: determination of structural, material, and mechanical properties. AB - This paper is devoted to the experimental determination of distinctive macroscopic structural (porosity, tortuosity, and permeability) and mechanical (Biot-Willis elastic constants) properties of human trabecular bones. Then, the obtained data may serve as input parameters for modeling wave propagation in cancellous bones using Biot's theory. The goal of the study was to obtain experimentally those characteristics for statistically representative group of human bones (35 specimens) obtained from a single skeletal site (proximal femur). The structural parameters were determined using techniques devoted to the characterization of porous materials: electrical spectroscopy, water permeametry, and microcomputer tomography. The macroscopic mechanical properties, Biot-Willis elastic constants, were derived based on the theoretical consideration of Biot's theory, micromechanical statistical models, and experimental results of ultrasonic studies for unsaturated cancellous bones. Our results concerning structural parameters are consistent with the data presented by the other authors, while macroscopic mechanical properties measured within our studies are situated between the other published data. The discrepancies are mainly attributed to different mechanical properties of the skeleton frame, due to strong structural anisotropy varying from site to site. The results enlighten the difficulty to use Biot's theory for modeling wave propagation in cancellous bone, implying necessity of individual evaluation of input parameters. PMID- 18397045 TI - Automated species recognition of antbirds in a Mexican rainforest using hidden Markov models. AB - Behavioral and ecological studies would benefit from the ability to automatically identify species from acoustic recordings. The work presented in this article explores the ability of hidden Markov models to distinguish songs from five species of antbirds that share the same territory in a rainforest environment in Mexico. When only clean recordings were used, species recognition was nearly perfect, 99.5%. With noisy recordings, performance was lower but generally exceeding 90%. Besides the quality of the recordings, performance has been found to be heavily influenced by a multitude of factors, such as the size of the training set, the feature extraction method used, and number of states in the Markov model. In general, training with noisier data also improved recognition in test recordings, because of an increased ability to generalize. Considerations for improving performance, including beamforming with sensor arrays and design of preprocessing methods particularly suited for bird songs, are discussed. Combining sensor network technology with effective event detection and species identification algorithms will enable observation of species interactions at a spatial and temporal resolution that is simply impossible with current tools. Analysis of animal behavior through real-time tracking of individuals and recording of large amounts of data with embedded devices in remote locations is thus a realistic goal. PMID- 18397047 TI - Adaptively biased molecular dynamics for free energy calculations. AB - We present an adaptively biased molecular dynamics (ABMD) method for the computation of the free energy surface of a reaction coordinate using nonequilibrium dynamics. The ABMD method belongs to the general category of umbrella sampling methods with an evolving biasing potential and is inspired by the metadynamics method. The ABMD method has several useful features, including a small number of control parameters and an O(t) numerical cost with molecular dynamics time t. The ABMD method naturally allows for extensions based on multiple walkers and replica exchange, where different replicas can have different temperatures and/or collective variables. This is beneficial not only in terms of the speed and accuracy of a calculation, but also in terms of the amount of useful information that may be obtained from a given simulation. The workings of the ABMD method are illustrated via a study of the folding of the Ace GGPGGG-Nme peptide in a gaseous and solvated environment. PMID- 18397048 TI - Correlation holes for the helium dimer. AB - We have investigated the radial electron pair probability distributions (REPPDs) of the helium dimer within the Piris natural orbital functional (PNOF) theory. The analytical formulas to evaluate intracule densities, Fermi, Coulomb, and total correlation holes using our reconstruction functional PNOF-2 [J. Chem. Phys. 126, 214103 (2007)] are derived. The Lowdin's Coulomb holes from PNOF-2 and full configuration interaction calculations are analyzed showing a very similar behavior. New definitions of the Coulomb and Fermi holes based on the cumulant expansion of the two-particle reduced density matrix are presented. The holes are defined in terms of the exact one-particle reduced density matrix and the two particle cumulant without any reference to the Hartree-Fock state. Through these definitions, we analyze separately the contribution of each component to the total REPPD at several values of the internuclear distance. A straight connection between the Coulomb hole and dispersion interactions is observed. PMID- 18397049 TI - Two-surface Monte Carlo with basin hopping: quantum mechanical trajectory and multiple stationary points of water cluster. AB - The efficiency of the two-surface monte carlo (TSMC) method depends on the closeness of the actual potential and the biasing potential used to propagate the system of interest. In this work, it is shown that by combining the basin hopping method with TSMC, the efficiency of the method can be increased by several folds. TSMC with basin hopping is used to generate quantum mechanical trajectory and large number of stationary points of water clusters. PMID- 18397050 TI - Linear-scaling fixed-node diffusion quantum Monte Carlo: accounting for the nodal information in a density matrix-based scheme. AB - A reformulation of the fixed-node diffusion quantum Monte Carlo method (FN-DQMC) in terms of the N-particle density matrix is presented, which allows us to reduce the computational effort to linear for the evaluation of the local energy. The reformulation is based on our recently introduced density matrix-based approach for a linear-scaling variational QMC method [J. Kussmann et al., Phys. Rev. B. 75, 165107 (2007)]. However, within the latter approach of using the positive semi-definite N-particle trial density (rhoN T(R)=mid R:Psi(T)(R)mid R:(2)), the nodal information of the trial function is lost. Therefore, a straightforward application to the FN-DQMC method is not possible, in which the sign of the trial function is usually traced in order to confine the random walkers to their nodal pockets. As a solution, we reformulate the FN-DQMC approach in terms of off diagonal elements of the N-particle density matrix rhoN T(R;R'), so that the nodal information of the trial density matrix is obtained. Besides all-electron moves, a scheme to perform single-electron moves within N-PDM QMC is described in detail. The efficiency of our method is illustrated for exemplary calculations. PMID- 18397051 TI - Auxiliary density perturbation theory. AB - A new approach, named auxiliary density perturbation theory, for the calculation of second energy derivatives is presented. It is based on auxiliary density functional theory in which the Coulomb and exchange-correlation potentials are expressed by auxiliary function densities. Different to conventional coupled perturbed Kohn-Sham equations the perturbed density matrix is obtained noniteratively by solving an inhomogeneous equation system with the dimension of the auxiliary function set used to expand the auxiliary function density. A prototype implementation for the analytic calculation of molecular polarizabilities is presented. It is shown that the polarizabilities obtained with the newly developed auxiliary density perturbation approach match quantitative with the ones from standard density functional theory if augmented auxiliary function sets are used. The computational advantages of auxiliary density perturbation theory are discussed, too. PMID- 18397052 TI - Optimization methods for finding minimum energy paths. AB - A comparison of chain-of-states based methods for finding minimum energy pathways (MEPs) is presented. In each method, a set of images along an initial pathway between two local minima is relaxed to find a MEP. We compare the nudged elastic band (NEB), doubly nudged elastic band, string, and simplified string methods, each with a set of commonly used optimizers. Our results show that the NEB and string methods are essentially equivalent and the most efficient methods for finding MEPs when coupled with a suitable optimizer. The most efficient optimizer was found to be a form of the limited-memory Broyden-Fletcher-Goldfarb-Shanno method in which the approximate inverse Hessian is constructed globally for all images along the path. The use of a climbing-image allows for finding the saddle point while representing the MEP with as few images as possible. If a highly accurate MEP is desired, it is found to be more efficient to descend from the saddle to the minima than to use a chain-of-states method with many images. Our results are based on a pairwise Morse potential to model rearrangements of a heptamer island on Pt(111), and plane-wave based density functional theory to model a rollover diffusion mechanism of a Pd tetramer on MgO(100) and dissociative adsorption and diffusion of oxygen on Au(111). PMID- 18397053 TI - Dissipation of classical energy in nonlinear quantum systems. AB - We show using two simple nonlinear quantum systems that the infinite set of quantum dynamical variables, as introduced in quantized Hamilton dynamics [O. V. Prezhdo and Y. V. Pereverzev, J. Chem. Phys. 113, 6557 (2000)], behave as a thermostat with respect to the finite number of classical variables. The coherent classical component of the evolution decays by coupling to the chaotic quantum reservoir. The classical energy, understood as the part of system energy expressible through the average values of coordinates and momenta, is transferred to the quantum energy expressible through the higher moments of coordinates and momenta and other quantum variables. At long times, the classical variables reach equilibrium, and the classical energy fluctuates around the equilibrium value. These phenomena are illustrated with the exactly solvable Jaynes-Cummings model and a nonlinear oscillator. PMID- 18397054 TI - Projected gradient algorithms for Hartree-Fock and density matrix functional theory calculations. AB - We present projected gradient algorithms designed for optimizing various functionals defined on the set of N-representable one-electron reduced density matrices. We show that projected gradient algorithms are efficient in minimizing the Hartree-Fock or the Muller-Buijse-Baerends functional. On the other hand, they converge very slowly when applied to the recently proposed BBk (k=1,2,3) functionals [O. Gritsenko et al., J. Chem. Phys. 122, 204102 (2005)]. This is due to the fact that the BBk functionals are not proper functionals of the density matrix. PMID- 18397055 TI - Boltzmann bias grand canonical Monte Carlo. AB - We derive an efficient method for the insertion of structured particles in grand canonical Monte Carlo simulations of adsorption in very confining geometries. We extend this method to path integral simulations and use it to calculate the isotherm of adsorption of hydrogen isotopes in narrow carbon nanotubes (two dimensional confinement) and slit pores (one-dimensional confinement) at the temperatures of 20 and 77 K, discussing its efficiency by comparison to the standard path integral grand canonical Monte Carlo algorithm. We use this algorithm to perform multicomponent simulations in order to calculate the hydrogen isotope selectivity for adsorption in narrow carbon nanotubes and slit pores at finite pressures. The algorithm described here can be applied to the study of adsorption of real oligomers and polymers in narrow pores and channels. PMID- 18397056 TI - Benchmarks for electronically excited states: CASPT2, CC2, CCSD, and CC3. AB - A benchmark set of 28 medium-sized organic molecules is assembled that covers the most important classes of chromophores including polyenes and other unsaturated aliphatic compounds, aromatic hydrocarbons, heterocycles, carbonyl compounds, and nucleobases. Vertical excitation energies and one-electron properties are computed for the valence excited states of these molecules using both multiconfigurational second-order perturbation theory, CASPT2, and a hierarchy of coupled cluster methods, CC2, CCSD, and CC3. The calculations are done at identical geometries (MP26-31G*) and with the same basis set (TZVP). In most cases, the CC3 results are very close to the CASPT2 results, whereas there are larger deviations with CC2 and CCSD, especially in singlet excited states that are not dominated by single excitations. Statistical evaluations of the calculated vertical excitation energies for 223 states are presented and discussed in order to assess the relative merits of the applied methods. CC2 reproduces the CC3 reference data for the singlets better than CCSD. On the basis of the current computational results and an extensive survey of the literature, we propose best estimates for the energies of 104 singlet and 63 triplet excited states. PMID- 18397057 TI - Self-adaptive enhanced sampling in the energy and trajectory spaces: accelerated thermodynamics and kinetic calculations. AB - Here, we introduce a simple self-adaptive computational method to enhance the sampling in energy, configuration, and trajectory spaces. The method makes use of two strategies. It first uses a non-Boltzmann distribution method to enhance the sampling in the phase space, in particular, in the configuration space. The application of this method leads to a broad energy distribution in a large energy range and a quickly converged sampling of molecular configurations. In the second stage of simulations, the configuration space of the system is divided into a number of small regions according to preselected collective coordinates. An enhanced sampling of reactive transition paths is then performed in a self adaptive fashion to accelerate kinetics calculations. PMID- 18397058 TI - State-selected imaging of HCCO radical photodissociation dynamics. AB - We present a dc sliced ion imaging study of HCCO radical photodissociation to CH and CO at 230 nm. The measurements were made using a two-color reduced Doppler probe strategy. The CO rotational distribution was consistent with a Boltzmann distribution at 3500 K. Using the dc slice ion imaging approach, we obtained CO images for various rotational levels of CO (v=0). The results are largely consistent with earlier work, albeit with a significant 0.9 eV peak seen previously in the translational energy distributions absent in our state-selected imaging study. PMID- 18397059 TI - Transport of O+ through argon gas. AB - New experimental and theoretical results are presented that address the movement of O+ ions through argon gas. On the experimental front, improved ion mobility results are presented. These results confirm the presence of the oft-cited mobility minimum as a function of electrostatic field strength at room temperature. On the theoretical side, high-level ab initio potential energy curves are calculated for the Ar-O+ system and, from these, transport properties are calculated and compared to experiment. A crossing between the lowest 2Pi curve and the ground state 4Sigma(-) curve near the minimum of each potential becomes an avoided crossing on the inclusion of spin-orbit coupling. It is shown that the more appropriate potential for the description of the motion of O+(4S degree) through Ar at the energies of interest is the diabatic potential, neglecting fine structure. By using an improved 4Sigma(-) potential, agreement with the mobility measurements is obtained for low and intermediate electrostatic field strengths, although small discrepancies remain for high field strengths. The appropriate choice of diabatic or adiabatic potentials is also considered for related systems of interest: He-O+, Ne-O+, and Rg-O(-) (Rg=He,Ne,Ar). PMID- 18397060 TI - Steering dissociation of Br2 molecules with two femtosecond pulses via wave packet interference. AB - The dissociation dynamics of Br2 molecules induced by two femtosecond pump pulses are studied based on the calculation of time-dependent quantum wave packet. Perpendicular transition from X 1Sigma g+ to A 3Pi 1u+ and 1Pi 1u+ and parallel transition from X 1Sigma g+ to B 3Pi 0u+, involving two product channels Br (2P3/2)+Br (2P3/2) and Br (2P3/2)+Br* (2P1/2), respectively, are taken into account. Two pump pulses create dissociating wave packets interfering with each other. By varying laser parameters, the interference of dissociating wave packets can be controlled, and the dissociation probabilities of Br2 molecules on the three excited states can be changed to different degrees. The branching ratio of Br*/(Br+Br*) is calculated as a function of pulse delay time and phase difference. PMID- 18397061 TI - Including anharmonicity in the calculation of rate constants. II. The OH+H2- >H2O+H reaction. AB - A recently developed method for calculating anharmonic vibrational energy levels at nonstationary points along a reaction path that is based on second-order perturbation theory in curvilinear coordinates is combined with variational transition state theory with semiclassical multidimensional tunneling approximations to calculate thermal rate constants for the title reaction. Two different potential energy surfaces were employed for these calculations, an improved version of the author's surface 5 and the WSLFH surface of Wu et al. [J. Chem. Phys. 113, 3150 (2000)]. We present detailed comparisons of rate constants computed for the two surfaces with and without anharmonicity and with various approximations for incorporating tunneling along the reaction path. The results for this system are quite sensitive to the surface employed, the choice of coordinates (curvilinear versus rectilinear), and the inclusion of anharmonicity. A comparison with experiment provides information on the accuracy of these surfaces. PMID- 18397062 TI - Properties and long range interactions of the calcium atom. AB - The properties of a number of states of calcium are determined from a large basis configuration interaction calculation. The main focus is on the polarizabilities of the low lying states (the 4s2 1Se, 4s3d 1,3De, 4s4p 1,3Po, and 4s5s 1,3Se states) and the dispersion interactions of those states with the calcium ground state, the hydrogen atom, and the rare gases. PMID- 18397063 TI - Electron emission from laser-heated fullerene dianions: probing the repulsive Coulomb barrier. AB - Electron detachment from fullerene dianions Cm2- (m=76,84) stored in a room temperature Penning trap was probed upon pulsed laser excitation at wavelengths of 355 and 532 nm. The fraction of Cm(2-) surviving trapping times exceeding tens of milliseconds under UHV conditions, as well as the fraction of singly charged anions Cm(-) generated were recorded as a function of the laser fluence. Analysis by means of Poisson statistics yields absolute absorption cross sections and the number of photons necessary to induce the detachment. The cross sections obtained are in good agreement with the literature values. By describing the electron detachment as a statistical unimolecular process, we deduce effective activation energies from the number of photons required. These energies are compared to the sum of the second electron affinity and the Coulomb barrier height as calculated from an electrostatic charging model. PMID- 18397064 TI - Spin-orbit effects in the photoabsorption of WAu12 and MoAu12: a relativistic time dependent density functional study. AB - The electronic structure of both WAu12 and MoAu12 has been calculated at the density functional theory (DFT) level, employing the zero order regular approximation at the scalar relativistic level and including a spin-orbit coupling. The effect of the inclusion of the spin-orbit coupling is discussed, and the differences assigned to the nature of the encaged atom (W or Mo) are identified. Then, the excitation spectra of both clusters are calculated at the time-dependent DFT level, also in this case at both scalar relativistic and spin orbit levels. The inclusion of spin-orbit coupling is mandatory for an accurate description in the low energy region. At higher energy, where the density of states is higher, the convoluted intensity can be properly described already at the scalar relativistic level. The consequences of the spin-orbit coupling on the excitation spectrum of the clusters indicate that while in WAu12 the lowest excitations are essentially shifted in energy with respect to the scalar relativistic results, in MoAu12, a dramatic splitting in many lines is actually predicted, revealing a quite different behavior of the two clusters. PMID- 18397065 TI - Dissociative recombination of OPCl+ and OPCl2+: pushing the upper mass limit at CRYRING. AB - The dissociative recombination of OPCl+ and OPCl2+ has been studied at the storage ring CRYRING. The rate constants as a function of electron temperature have been derived to be 7.63 x 10(-7)(Te/300)(-0.89) and >1.2 x 10(-6)(Te/300)( 1.22) cm3s(-1), respectively. The lower limit quoted for the latter rate constant reflects the experimental inability to detect all of the reaction products. The branching fractions from the reaction have been measured for OPCl+ at approximately 0 eV interaction energy and are determined to be N(O+P+Cl)=(16+/ 7)%, N(O+PCl)=(16+/-3)% and N(OP+Cl)=(68+/-5)%. These values have been obtained assuming that the rearrangement channel forming P+ClO is negligible, and ab initio calculations using GAUSSIAN03 are presented for the ion structures and energetics to support such an assumption. Finally, the limitations to using heavy ion storage rings such as CRYRING for studies into the dissociative recombination of large singly charged molecular ions are discussed. PMID- 18397066 TI - Photofragmentation of SiF4 upon Si 2p and F 1s core excitation: cation and anion yield spectroscopy. AB - We have studied the fragmentation dynamics of core-excited SiF4 by means of soft x-ray photoexcitation and partial positive and negative ion yield measurements around the Si L2,3-shell and F K-shell ionization thresholds. All detectable ionic fragments are reported and we observe significant differences between the various partial ion yields near the Si 2p threshold. The differences are similar to our previous results from CH3Cl showing more extended fragmentation in correspondence to transitions to Rydberg states. At variance with smaller systems, we observe negative ion production in the shape resonance region. This can be related to the possibility in a relatively large system to dissipate positive charge over several channels. PMID- 18397067 TI - Molecular beam resonant two-photon ionization study of caffeine and its hydrated clusters. AB - We investigated electronically excited states of caffeine and its 1:1 complex with water by using resonant two-photon ionization (R2PI) and UV-UV hole-burning techniques. Strong vibronic coupling between a pair of close-lying pi-pi* and n pi* transitions is proposed to be responsible for the broad spectral feature observed. By comparing the experimental results with those of theoretical calculations, both the O-bonded and N-bonded forms were suggested to be initially produced for the 1:1 complex. The electronic transitions of the O-bonded complex were blueshifted in the R2PI spectrum. For the N-bonded complex, the excited state undergoes an ultrafast decay process, followed by dissociation on a repulsive potential energy surface, which gives rise to a characteristically anomalous cluster distribution in nanosecond experiments. PMID- 18397068 TI - Product state resolved excitation spectroscopy of He-, Ne-, and Ar-Br2 linear isomers: experiment and theory. AB - Valence excitation spectra for the linear isomers of He-, Ne-, and Ar-Br2 are reported and compared to a two-dimensional simulation using the currently available potential energy surfaces. Excitation spectra from the ground electronic state to the region of the inner turning point of the Rg-Br2 (B,nu') stretching coordinate are recorded while probing the asymptotic Br2 (B,nu') state. Each spectrum is a broad continuum extending over hundreds of wavenumbers, becoming broader and more blueshifted as the rare gas atom is changed from He to Ne to Ar. In the case of Ne-Br2, the threshold for producing the asymptotic product state reveals the X-state linear isomer bond energy to be 71+/-3 cm(-1). The qualitative agreement between experiment and theory shows that the spectra can be correctly regarded as revealing the one-atom solvent shifts and also provides new insight into the one-atom cage effect on the halogen vibrational relaxation. The measured spectra provide data to test future ab initio potential energy surfaces in the interaction of rare gas atoms with the halogen valence excited state. PMID- 18397069 TI - The homogeneous nucleation of 1-pentanol in a laminar flow diffusion chamber: the effect of pressure and kind of carrier gas. AB - The influence of total pressure and kind of carrier gas on homogeneous nucleation rates of 1-pentanol was investigated using experimental method of laminar flow diffusion chamber in this study. Two different carrier gases (helium and argon) were used in the total pressure range from 50 to 400 kPa. Nucleation temperatures ranged from 265 to 290 K for 1-pentanol-helium and from 265 to 285 K for 1 pentanol-argon. Nucleation rates varied between 10(1) and 10(6) cm(-3) s(-1) for 1-pentanol-helium and between 10(2) and 10(5) cm(-3) s(-1) for 1-pentanol-argon. Both positive and slight negative pressure effects were observed depending on temperature and carrier gas. The trend of pressure effect was found similar for both carrier gases. Error analysis on thermodynamic properties was conducted, and the lowering of surface tension due to adsorption of argon on nucleated droplets was estimated. A quantitative overview of pressure effect is provided. PMID- 18397070 TI - Interactions of the 3ppi u c1Pi u(v=2) Rydberg-complex member in isotopic N2. AB - The 3ppi u c1Pi u-X 1Sigmag+(2,0) Rydberg and b' 1Sigmau+-X 1Sigmag+(7,0) valence transitions of 14N2, 14N15N, and 15N2 are studied using laser-based 1 extreme ultraviolet (XUV)+1' UV two-photon-ionization spectroscopy, supplemented by synchrotron-based hotoabsorption measurements in the case of 14N2. For each isotopomer, effective rotational interactions between the c(v=2) and b'(v=7) levels are found to cause strong Lambda-doubling in c(v=2) and dramatic P/R branch intensity anomalies in the b'-X(7,0) band due to the effects of quantum interference. Local perturbations in energy and predissociation line width for the c(v=2) Rydberg level are observed and attributed to a spin-orbit interaction with the crossing, short-lived C 3Pi u(v=17) valence level. PMID- 18397071 TI - On the computation and contribution of conductivity in molecular ionic liquids. AB - In this study we present the results of the molecular dynamics simulation of the ionic liquids: 1-butyl-3-methyl-imidazolium tetrafluoroborate and trifluoromethylacetate as well as 1-ethyl-3-methyl-imidazolium dicyanamide. Ionic liquids are characterized by both a molecular dipole moment and a net charge. Thus, in contrast to a solution of simple ions in a (non-) polar solvent, rotational and translational effects influence the very same molecule. This study works out the theoretical framework necessary to compute the conductivity spectrum and its low frequency limit of ionic liquids. Merging these computed conductivity spectra with previous simulation results on the dielectric spectra of ionic liquids yields the spectrum of the generalized dielectric constant, which may be compared to experiments. This spectrum was calculated for the three ionic liquids over six orders of magnitude in frequency ranging from 10 MHz to 50 THz. The role of rotation and translation and their coupling term on the generalized dielectric constant is discussed in detail with a special emphasis on the zero-frequency limit. Thereby, the frequency dependence of the cross correlation between the collective rotational dipole moment and the current is discussed. PMID- 18397072 TI - Solid-state NMR covariance of homonuclear correlation spectra. AB - Direct covariance NMR spectroscopy, which does not involve a Fourier transformation along the indirect dimension, is demonstrated to obtain homonuclear correlation two-dimensional (2D) spectra in the solid state. In contrast to the usual 2D Fourier transform (2D-FT) NMR, in a 2D covariance (2D Cov) spectrum the spectral resolution in the indirect dimension is determined by the resolution along the detection dimension, thereby largely reducing the time consuming indirect sampling requirement. The covariance method does not need any separate phase correction or apodization along the indirect dimension because it uses those applied in the detection dimension. We compare in detail the specifications obtained with 2D-FT and 2D-Cov, for narrow and broad resonances. The efficiency of the covariance data treatment is demonstrated in organic and inorganic samples that are both well crystallized and amorphous, for spin -1/2 nuclei with 13C, 29Si, and 31P through-space or through-bond homonuclear 2D correlation spectra. In all cases, the experimental time has been reduced by at least a factor of 10, without any loss of resolution and signal to noise ratio, with respect to what is necessary with the 2D-FT NMR. According to this method, we have been able to study the silicate network of glasses by 2D NMR within reasonable experimental time despite the very long relaxation time of the 29Si nucleus. The main limitation of the 2D-Cov data treatment is related to the introduction of autocorrelated peaks onto the diagonal, which does not represent any actual connectivity. PMID- 18397073 TI - Monte Carlo simulations of fluids whose particles interact with a logarithmic potential. AB - Monte Carlo simulations of a model fluid in which the particles interact via a continuous potential that has a logarithmic divergence at a pair separation of sigma, which we introduced in J. G. Powles et al., Proc. R. Soc. London, Ser. A 455, 3725 (1999), have been carried out. The potential has the form, phi(r)= epsilon ln(fr), where epsilon sets the energy scale and fr=1-(sigma/r)m. The value of m chosen was 12 but the qualitative trends depend only weakly on the value of m, providing it is greater than 3. The potential is entirely repulsive and has a logarithmic divergence as approximately -ln(r/sigma-1) in the r-->sigma limit. Predictions of the previous paper that the internal energy can be computed at all temperatures using the standard statistical mechanics formula for continuous potentials are verified here. The pressure can be calculated using the usual virial expression for continuous potentials, although there are practical limitations in resolving the increasingly important contribution from the r- >sigma limit at reduced temperatures greater than approximately 5. The mean square force F2 and infinite frequency shear Ginfinity and bulk Kinfinity moduli are only finite for T*=kBT/epsilon<1. The logarithmic fluid's physical properties become increasingly more like that of the hard sphere fluid with increasing temperature, showing a sharp transition in the behavior of the mean square force and infinite frequency elastic constants at T*=1. The logarithmic fluid is shown to exhibit a solid-fluid phase transition. PMID- 18397074 TI - Ordering layers of [bmim][PF6] ionic liquid on graphite surfaces: molecular dynamics simulation. AB - Microscopic structures of room temperature ionic liquid (IL) [bmim][PF6] on hydrophobic graphite surfaces have been studied in detail by molecular dynamics simulation. It is clearly shown that both the mass and electron densities of the surface adsorbed ionic liquid are oscillatory, and the first peak adjacent to the graphite surface is considerably higher than others, corresponding to a solidlike IL bottom layer of 6 angstroms thick. Three IL layers are indicated between the graphite surface and the inner bulk IL liquid. The individually simulated properties of single-, double-, and triple-IL layers on the graphite surface are very similar to those of the layers between the graphite surface and the bulk liquid, indicating an insignificant effect of vapor-IL interface on the ordered IL layers. The simulation also indicates that the imidazolium ring and butyl tail of the cation (bmim+) of the IL bottom layer lie flat on the graphite surface. PMID- 18397075 TI - Treatment of charged solutes in three-dimensional integral equation theory. AB - Periodicity artifacts, which occur within three-dimensional reference interaction site model integral equation theory for net-charged solute systems, are analyzed and corrected by means of a renormalization procedure for long range interactions. The method is formulated for solute-solvent and solute-solute variants of the theory. Both dielectric and electrolyte solvents are considered. Comparison of the results for atomic ions with one-dimensional reference computations shows that structural and thermodynamic artifacts are efficiently removed. PMID- 18397076 TI - Nitrile and thiocyanate IR probes: quantum chemistry calculation studies and multivariate least-square fitting analysis. AB - Hydration effects on the C[Triple Bond]N stretching mode frequencies of MeCN and MeSCN are investigated by carrying out ab initio calculations for a number of MeCN-water and MeSCN-water complexes with varying number of water molecules. It is found that the CN frequency shift induced by the hydrogen-bonding interactions with water molecules originate from two different ways to form hydrogen bonds with the nitrogen atom of the CN group. Considering the MeCN- and MeSCN-water cluster calculation results as databases, we first examined the validity of vibrational Stark effect relationship between the CN frequency and the electric field component parallel to the CN bond and found no strong correlation between the two. However, taking into account of additional electric field vector components is a simple way to generalize the vibrational Stark theory for the nitrile chromophore. Also, the electrostatic potential calculation method has been proposed and examined in detail. It turned out that the interactions of water molecules with nitrogen atom's lone pair orbital and with nitrile pi orbitals can be well described by the electrostatic potential calculation method. The present computational results will be of use to quantitatively simulate various linear and nonlinear vibrational spectra of nitrile compounds in solutions. PMID- 18397077 TI - Depletion potential in the infinite dilution limit. AB - The depletion force and depletion potential between two in principle unequal "big" hard spheres embedded in a multicomponent mixture of "small" hard spheres are computed using the rational function approximation method for the structural properties of hard-sphere mixtures [S. B. Yuste, A. Santos, and M. Lopez de Haro, J. Chem. Phys. 108, 3683 (1998)]. The cases of equal solute particles and of one big particle and a hard planar wall in a background monodisperse hard-sphere fluid are explicitly analyzed. An improvement over the performance of the Percus Yevick theory and good agreement with available simulation results are found. PMID- 18397078 TI - On the corresponding states law of the Yukawa fluid. AB - We have analyzed the currently available simulation results as well as performed some additional Monte Carlo simulation for the hard-core attractive Yukawa fluid in order to study its corresponding state behavior. We show that the values of reduced surface tension map onto the master curve and a universal equation of state can be obtained in the wide range of the attractive Yukawa tail length after a certain rescaling of the number density. Some comparisons with other nonconformal potentials are presented and discussed. PMID- 18397079 TI - Evolution of the vibrational spectrum of ammonia from single molecule to bulk. AB - Ammonia clusters (NH3)n (n=2-10(4)) have been assembled inside helium droplets and studied via infrared laser spectroscopy. The studied spectral range of 3100 3500 cm(-1) covers the nu1 and nu3 fundamental stretching bands as well as the 2nu4 overtone of the bend of ammonia molecules. The results show strong coupling of the 2nu4 overtone with the fundamental vibrations for all cluster sizes except dimers. The intensity of the nu3 band relative to the total intensity in the spectrum increases from about 30% to about 80% upon increase of the average cluster size from n=5 to n=10(4). We attributed this effect to the concomitant decrease in the fraction of the surface molecules. The results indicate that ammonia clusters obtained in He droplets have a compact structure and that inner molecules in the clusters have similar hydrogen-bonded coordination as in the crystalline form of ammonia. This surprising result is ascribed to a directionality of the hydrogen bond, which guides the low temperature growth of the cluster in He droplets. PMID- 18397080 TI - Effect of polydispersity on the relative stability of hard-sphere crystals. AB - By extending the nonequilibrium potential refinement algorithm and lattice switch method to the semigrand ensemble, the semigrand potentials of the fcc and hcp structures of polydisperse hard-sphere crystals are calculated with the bias sampling scheme. The result shows that the fcc structure is more stable than the hcp structure for polydisperse hard-sphere crystals below the terminal polydispersity. PMID- 18397081 TI - Anionic microsolvation in helium droplets: OH- (He)N structures from classical and quantum calculations. AB - Diffusion Monte Carlo calculations are carried out for clusters of OH- (1Sigma+) with N 4He atoms, N varying up to 15, while classical configurations from a genetic algorithm optimization are obtained up to N=20. The overall interaction potential is assembled from ab initio data for the partners using the sum-of potentials scheme. In contrast with the cationic dopants' behavior, the results indicate a very marked spatial delocalization and quantum features of the solvent adatoms surrounding the anionic impurity, thus making classical calculations of solvent's spatial locations of only limited use. In spite of the generally known repulsive interaction of negative charges in He droplets, the calculations show that this polar molecular anion is solvated by a liquidlike solvent layer, reminiscent of what happens in pure helium droplets. PMID- 18397082 TI - Vibrational dynamics of amorphous beryllium hydride and lithium beryllium hydrides. AB - The vibrational density of states of amorphous beryllium hydride (a-BeH2) and lithium beryllium hydrides have been studied using inelastic neutron scattering, infrared, and Raman spectroscopies. The positions of the symmetrical (120-180 meV) and antisymmetrical (200-260 meV) Be-H stretching modes and those of the H Be-H bending mode (50-120 meV) have been determined and the results discussed and compared with recent theoretical calculations. With the addition of lithium to the beryllium hydride network, the vibrational bands are shifted to lower energies, indicating a less rigid network. PMID- 18397083 TI - The vanishing limit of the square-well fluid: the adhesive hard-sphere model as a reference system. AB - We report a simulation study of the gas-liquid critical point for the square-well potential, for values of well width delta as small as 0.005 times the particle diameter sigma. For small delta, the reduced second virial coefficient at the critical point B2*c is found to depend linearly on delta. The observed weak linear dependence is not sufficient to produce any significant observable effect if the critical temperature Tc is estimated via a constant B2*c assumption, due to the highly nonlinear transformation between B2*c and Tc. This explains the previously observed validity of the law of corresponding states. The critical density rho c is also found to be constant when measured in units of the cube of the average distance between two bonded particles (1+0.5 delta)sigma. The possibility of describing the delta-->0 dependence with precise functional forms provides improved accurate estimates of the critical parameters of the adhesive hard-sphere model. PMID- 18397084 TI - Characterization of mono- and divacancy in fcc and hcp hard-sphere crystals. AB - We determine and compare the thermodynamic properties of mono- and divacancies in the face-centered-cubic and hexagonal-close-packed hard-sphere crystals via a modified grand canonical ensemble. Widom-type particle insertion was employed to estimate the free energy of formation of mono- and divacancies, and the results are supported by an alternative approach, which quantifies the entropy gain of the neighbor particles. In hcp crystal, we found a strong anisotropy in the orientational distribution of vacancies and observe an eightfold increase in the number of divacancies in the hexagonal plane compared to the one in the out of plane at highest density of interest. This phenomenon is induced by the different arrangement and behavior of the shared nearest neighbor particles, which are located at the same distance from each vacant site in divacancy. The effect of divacancies on the free energy is to reduce that of the hcp crystal relative to the fcc by around 7 x 10(-6)kBT at melting. PMID- 18397085 TI - Lone pair versus bonding pair electrons: the mechanism of electronic polarization of water in the presence of positive ions. AB - It is commonly accepted that the water molecules in the first solvation shell of a positive ion are strongly polarized because of an elongation of the oxygen lone pair orbitals along the ion-oxygen direction and this is commonly considered the dominant effect. Recent experimental and theoretical works have instead suggested that this is not the dominant aspect and that the problem is by far more complicated. Consistent with the picture given above, here we show that, in particular, an equally important role into the polarization process is played by the bonding pair electrons located along the internal oxygen-hydrogen bond. We also provide some arguments which suggest that the main reason of such a behavior is due to the distortion of the molecular orbitals caused by the interaction between non-hydrogen-bonded water molecules in the first solvation shell of the ion. PMID- 18397086 TI - Use of the cage formation probability for obtaining approximate phase diagrams. AB - In this work, we introduce the idea of cage formation probability, defined by considering the angular space needed by a particle in order to leave a cage given an average distance to its neighbors. Considering extreme fluctuations, two phases appear as a function of the number of neighbors and their distances to a central one: Solid and fluid. This allows us to construct an approximated phase diagram based on a geometrical approach. As an example, we apply this probability concept to hard disks in two dimensions and hard spheres in three dimensions. The results are compared with numerical simulations using a Monte Carlo method. PMID- 18397087 TI - Structural motifs, mixing, and segregation effects in 38-atom binary clusters. AB - Thirty eight-atom binary clusters composed of elements from groups 10 and 11 of the Periodic Table mixing a second-row with a third-row transition metal (TM) (i.e., clusters composed of the four pairs: Pd-Pt, Ag-Au, Pd-Au, and Ag-Pt) are studied through a combined empirical-potential (EP)/density functional (DF) method. A "system comparison" approach is adopted in order to analyze a wide diversity of structural motifs, and the energy competition among different structural motifs is studied at the DF level for these systems, mainly focusing on the composition 24-14 (the first number refers to the second-row TM atom) but also considering selected motifs with compositions 19-19 (of interest for investigating surface segregation effects) and 32-6 (also 14-24 and 6-32 for the Pd-Au pair). The results confirm the EP predictions about the stability of crystalline structures at this size for the Au-Pd pair but with decahedral or mixed fivefold-symmetric/closed-packed structures in close competition with fcc motifs for the Ag-Au or Ag-Pt and Pd-Pt pairs, respectively. Overall, the EP description is found to be reasonably accurate for the Pd-Pt and Au-Pd pairs, whereas it is less reliable for the Ag-Au and Ag-Pt pairs due to electronic structure (charge transfer or directionality) effects. The driving force to core shell chemical ordering is put on a quantitative basis, and surface segregation of the most cohesive element into the core is confirmed, with the exception of the Ag-Au pair for which charge transfer effects favor the segregation of Au to the surface of the clusters. PMID- 18397088 TI - Ice nucleation on hydrophilic silicon. AB - We have used Fourier transform infrared spectroscopy to study thin water films on a hydrophilic silicon surface in the temperature range from 20 to -20 degrees C. Throughout that range, the spectra of the water adjacent to the silicon surface are consistent with that of bulk water near 25 degrees C. Thicker films (>1 microm) freeze at -11+/-1 degrees C. We reconcile the apparent paradox of a thin film of water which is quite liquidlike at a temperature where freezing of thicker films occurs by hypothesizing that the nucleation event in the thicker film is triggered by a critical ice embryo which forms at some small distance from the silicon surface, as opposed to in direct contact with it. PMID- 18397089 TI - Analytical solution for restricted diffusion in circular and spherical layers under inhomogeneous magnetic fields. AB - We propose an analytical solution for restricted diffusion of spin-bearing particles in circular and spherical layers in inhomogeneous magnetic fields. More precisely, we derive exact and explicit formulas for the matrix representing an applied magnetic field in the Laplacian eigenbasis and governing the magnetization evolution. For thin layers, a significant difference between two geometrical length scales (thickness and overall size) allows for accurate perturbative calculations. In these two-scale geometries, apparent diffusion coefficient (ADC) as a function of diffusion time exhibits a new region with a reduced but constant value. The emergence of this intermediate diffusion regime, which is analogous to the tortuosity regime in porous media, is explained in terms of the underlying Laplace operator eigenvalues. In general, regions with constant ADCs would be reminiscent of multiscale geometries, and their observation can potentially be used in experiments to detect the length scales by varying diffusion time. PMID- 18397090 TI - Effects of size constraint on water filling process in nanotube. AB - Molecular dynamics (MD) simulation and the potential of mean force (PMF) analysis are used to investigate the structural properties of water molecules near the end of nanotube for the whole process from the initial water filling up to the configuration stabilization inside the carbon nanotubes (CNTs). Numerical simulations showed that when a small-sized nanotube is immersed into the water bath, the size constraint will induce a prevailing orientation for the water molecule to diffuse into the tube and this effect can persist approximately 3.3 angstroms from the end of CNT. As the structure within the CNTs stabilizes, the ambient structural properties can indirectly reflect their corresponding properties inside the nanotube. Our results also showed that there exists a close correlation between the PMF analysis and the results of MD simulations, and the properties at nanometer scale are closely related to the size-constraint effect. PMID- 18397091 TI - Diffusion at the liquid-vapor interface. AB - Recently, the intrinsic sampling method has been developed in order to obtain, from molecular simulations, the intrinsic structure of the liquid-vapor interface that is presupposed in the classical capillary wave theory. Our purpose here is to study dynamical processes at the liquid-vapor interface, since this method allows tracking down and analyzing the movement of surface molecules, thus providing, with great accuracy, dynamical information on molecules that are "at" the interface. We present results for the coefficients for diffusion parallel and perpendicular to the liquid-vapor interface of the Lennard-Jones fluid, as well as other time and length parameters that characterize the diffusion process in this system. We also obtain statistics of permanence and residence time. The generality of our results is tested by varying the system size and the temperature; for the latter case, an existing model for alkali metals is also considered. Our main conclusion is that, even if diffusion coefficients can still be computed, the turnover processes, by which molecules enter and leave the intrinsic surface, are as important as diffusion. For example, the typical time required for a molecule to traverse a molecular diameter is very similar to its residence time at the surface. PMID- 18397092 TI - Kinetic study of the "surface explosion" phenomenon in the NO+CO reaction on Pt(100) through dynamic Monte Carlo simulation. AB - The extremely narrow production peak of N2 and CO2 which occurs in the reaction of NO+CO on Pt(100), a phenomenon known as "surface explosion," is studied using a dynamic Monte Carlo method on a square lattice at low pressure under isothermal conditions. This analysis incorporates recent experimental evidence obtained for the same reaction on a Rh(111) surface, which has shown that N2 production occurs either from the classical N+N recombination step or by the formation and successive decay of a (N-NO)* intermediary species. Moreover, the NO dissociation rate is inhibited by coadsorbed NO and CO molecules and is enhanced both by the presence of empty sites and adsorbed N atoms as nearest neighbors. These effects are taken into account in this study, along with the experimental adsorption, desorption, and diffusion rates of the reactants. The "explosive" phenomenon is analyzed through the evolution over time of an adsorbed NO+CO monolayer at a fixed temperature of 400 K. Furthermore, as the diffusion processes of the adsorbates are included, cellular structures are observed. Our simulations show quantitative agreement in the position of maxima with those obtained through experiments using isothermal desorption mass spectroscopy. PMID- 18397093 TI - Geometric and electronic structure of lanthanide orthophosphate nanoparticles determined with X-rays. AB - The evolution of the geometric and electronic structures within the entire series of lanthanide orthophosphate nanoparticles ( approximately 2- approximately 5 nm) has been determined experimentally with X-ray diffraction and near edge X-ray absorption fine structure spectroscopy. In particular, the interplay between electronic structure, crystal morphology, and crystal phase has been systematically studied. A missing local order in the crystal structure accompanied by multiple ion sites in the nanoparticles was revealed to be due to the small crystal size and large surface contribution. All lanthanide ions were found to be in "3+" configuration and accommodated in three different crystallization states: the larger lanthanide ions (La, Ce, Pr, Nd, Sm) in the monoclinic phase, the smaller ones (Er, Tm, Yb, Lu) in the tetragonal phase, and the intermediate lanthanide ions (Eu, Gd, Tb, Dy, Ho) in a "mixed phase" between monoclinic and tetragonal phases. PMID- 18397094 TI - Binding characteristics of pyridine on Ag(110). AB - A combination of low-temperature scanning tunneling microscopy and density functional theory calculations was used to determine the binding characteristics of single pyridine molecules at a low coverage on a silver surface. The results indicated that pyridine binds to silver through the nitrogen atom in either a perpendicular or a parallel configuration with the latter structure being more prevalent. Both configurations are produced predominantly through electrostatic interaction between nitrogen and silver atoms. This is induced by charge redistribution in the pyridine molecule and nearby silver atoms upon pyridine adsorption. PMID- 18397095 TI - Self-organized transient facilitated atomic transport in Pt/Al(111). AB - During the course of atomic transport in a host material, impurity atoms need to surmount an energy barrier driven by thermodynamic bias or at ultralow temperatures by quantum tunneling. In the present article, we demonstrate using atomistic simulations that at ultralow temperature, transient interlayer atomic transport is also possible without tunneling when the Pt/Al(111) impurity/host system self-organizes itself spontaneously into an intermixed configuration. No such extremely fast athermal concerted process has been reported before at ultralow temperatures. The outlined novel transient atomic exchange mechanism could be of general validity. We find that the source of ultralow temperature heavy particle barrier crossing is intrinsic and no external bias is necessary for atomic intermixing and surface alloying in Pt/Al, although the dynamic barrier height is a few eV. The mechanism is driven by the local thermalization of the Al(111) surface in a self-organized manner arranged spontaneously by the system without any external stimulus. The core of the short lived thermalized region reaches the local temperature of approximately 1000 K (including a few tens of Al atoms), while the average temperature of the simulation cell is approximately 3 K. The transient facilitated intermixing process also takes place with repulsive impurity-host interaction potential leading to negative atomic mobility; hence, the atomic injection is largely independent of the strength of the impurity-surface interaction. We predict that similar exotic behavior is possible in other materials as well. PMID- 18397096 TI - Hydrogenation of CO on a silica surface: an embedded cluster approach. AB - The sequential addition of H atoms to CO adsorbed on a siliceous edingtonite surface is studied with an embedded cluster approach, using density functional theory for the quantum mechanical (QM) cluster and a molecular force field for the molecular mechanical (MM) cluster. With this setup, calculated QM/MM adsorption energies are in agreement with previous calculations employing periodic boundary conditions. The catalytic effect of the siliceous edingtonite (100) surface on CO hydrogenation is assessed because of its relevance to astrochemistry. While adsorption of CO on a silanol group on the hydroxylated surface did not reduce the activation energy for the reaction with a H atom, a negatively charged defect on the surface is found to reduce the gas phase barriers for the hydrogenation of both CO and H2C=O. The embedded cluster approach is shown to be a useful and flexible tool for studying reactions on (semi-)ionic surfaces and specific defects thereon. The methodology presented here could easily be applied to study reactions on silica surfaces that are of relevance to other scientific areas, such as biotoxicity of silica dust and geochemistry. PMID- 18397097 TI - Intrinsic property measurement of surfactant-templated mesoporous silica films using time-resolved single-molecule imaging. AB - Mesoporous silica membranes fabricated by the surfactant-templated sol-gel process have received attention because of the potential to prepare membranes with a narrow pore size distribution and ordering of the interconnected pores. Potential applications include ultrafiltration, biological separations and drug delivery, and separators in lithium-ion batteries. Despite advancements in synthesis and characterization of these membranes, a quantitative description of the membrane microstructure remains a challenge. Currently the membrane microstructure is characterized by the combination of results from several techniques, i.e., gas permeance testing, x-ray diffraction scanning electron microscopy, transmission electron microscopy, and permporometry. The results from these ensemble methods are then compiled and the data fitted to a particular flow model. Although these methods are very effective in determining membrane performance, general pore size distribution, and defect concentration, they are unable to monitor molecular paths through the membrane and quantitatively measure molecular interactions between the molecular specie and pore network. Single molecule imaging techniques enable optical measurements that probe materials on nanometer length scales through observation of individual molecules without the influence of averaging. Using single-molecule imaging spectroscopy, we can quantitatively characterize the interaction between the probe molecule and the interior of the pore within mesoporous silica membranes. This approach is radically different from typical membrane characterization methods in that it has the potential to spatially sample the underlying pore structure distribution, the surface energy, and the transport properties. Our hope is that this new fundamental knowledge can be quantitatively linked to both the preparation and the performance of membranes, leading to the advancement of membrane science and technology. Fluorescent molecules, 1,1-dioctadecyl-3,3,3,3-tetramethylindo carbocyanine perchlorate, used to interrogate the available free volume in their vicinity, were loaded into the mesoporous silica membranes at subnanomolar concentrations. The mesoporous silica films were prepared using a nonionic ethylene oxide-propylene oxide-ethylene oxide triblock copolymer surfactant, Pluronic P123, on single crystal silicon substrates using dip coating of a silica sol. Membranes were prepared resulting in an average pore diameter of approximately 5 nm as measured by helium, nitrogen permeance, and porosimetry. Fluorescent images and time transient experiments were recorded using a custom built single-molecule scanning confocal microscope at differing temperatures (10, 20, 30, 40, and 50 degrees C). Time-dependent polarization anisotropy was used to obtain the enthalpy of adsorption and Henry's law constant of the probe molecule. PMID- 18397098 TI - Adsorption geometry variation of 1,4-benzenedimethanethiol self-assembled monolayers on Au(111) grown from the vapor phase. AB - 1,4-benzenedimethanethiol was chemisorbed from the vapor phase onto Au(111). The chemisorption geometry, molecular orientation, and bonding properties were studied at different degrees of surface coverage by photoelectron spectroscopy, metastable deexcitation spectroscopy, and near-edge x-ray absorption fine structure spectroscopy at the carbon K edge. Two main chemisorption regimes were identified: at low coverage the molecules adopt a flat configuration, then, as the molecular density of the first layer increases, the reduction of the available chemisorption sites induces the newly bonded molecules to assume a vertical alignment, with only one of the sulphur head groups interacting with the substrate. Experimental results were interpreted on the basis of theoretical calculations that we performed on the free molecule concerning the molecular orbitals' density of states and simulated x-ray absorption. PMID- 18397099 TI - Interaction of methanol with amorphous solid water. AB - The interaction of methanol (MeOH) with amorphous solid water (ASW) composed of D2O molecules, prepared at 125 K on a polycrystalline Ag substrate, was studied with metastable-impact-electron spectroscopy, reflection-absorption infrared spectroscopy, and temperature-programmed desorption mass spectroscopy. In connection with the experiments, classical molecular dynamics (MD) simulations have been performed on a single CH3OH molecule adsorbed at the ice surface (T=190 K), providing further insights into the binding and adsorption properties of the molecule at the ice surface. Consistently with the experimental deductions and previous studies, MeOH is found to adsorb with the hydroxyl group pointing toward dangling bonds of the ice surface, the CH3 group being oriented upwards, slightly tilted with respect to the surface normal. It forms the toplayer up to the onset of the simultaneous desorption of D2O and MeOH. At low coverage the adsorption is dominated by the formation of two strong hydrogen bonds as evidenced by the MD results. During the buildup of the first methanol layer on top of an ASW film the MeOH-MeOH interaction via hydrogen-bond formation becomes of importance as well. The interaction of D2O with solid methanol films and the codeposition of MeOH and D2O were also investigated experimentally; these experiments showed that D2O molecules supplied to a solid methanol film become embedded into the film. PMID- 18397100 TI - Symmetry breaking in binary mixtures in closed nanoslits. AB - The symmetry breaking (SB) of the fluid density distribution (FDD) in closed nanoslits between two identical parallel solid walls described by Berim and Ruckenstein [J. Chem. Phys. 128, 024704 (2008)] for a single component fluid is examined for binary mixtures on the basis of a nonlocal canonical ensemble density functional theory. As in Monte Carlo simulations, the periodicity of the FDD in one of the lateral (parallel to the wall surfaces) directions, denoted as the x direction, was assumed. In the other lateral direction, y direction, the FDD was considered to be uniform. The molecules of the two components have different diameters and their Lennard-Jones interaction potentials have different energy parameters. It was found that depending on the average fluid density in the slit and mixture composition, SB can occur for both or none of the components but never for only one of them. In the direction perpendicular to the walls (h direction), the FDDs of both components can be asymmetrical about the middle plane between walls. In the x direction, the SB occurs as bumps and bridges enriched in one of the components, whereas the composition of the mixture between them is enriched in the other component. The dependence of the SB states on the length Lx of the FDD period at fixed average densities of the two components was examined for Lx in the range from 10 to 120 molecular diameters of the smaller size component. It was shown that for large Lx, the stable state of the system corresponds to a bridge. Because the free energy of that state decreases monotonically with increasing Lx, one can conclude that the real period is very large (infinite) and that a single bridge exists in the slit. PMID- 18397101 TI - Screening of hydrodynamic interactions in Brownian rod suspensions. AB - We present the details and results of a simulation study addressing the dynamics and rheology of rod suspensions over a wide regime of concentrations ranging from dilute to concentrated systems. Our study compares the results of two complementary simulation methods. The first method adapts a recently proposed explicit solvent simulation strategy and incorporates both hydrodynamical effects and steric interactions between the rod units. We compare the results of such a method with those obtained from a Brownian dynamics simulation approach which retains the steric interactions but neglects the effects of hydrodynamic interactions. Overall, our results in the context of the translational and rotational diffusivities are in agreement with the hydrodynamical predictions in the dilute regime and the corresponding results of the tube model and its extensions thereof in the semidilute regimes. The latter results suggest that effects of hydrodynamic interactions on the translational and rotational diffusivities are secondary relative to the steric interactions and at best lead only to a small correction to the results of the classical tube model. Our results in the context of linear viscoelasticity also broadly confirms the predictions of the tube model for the storage and loss moduli and allows us to extract for the first time the independent hydrodynamic and Brownian contributions to the zero shear viscosity. While the relative magnitudes of these contributions are consistent with the theoretical predictions, the quantitative magnitudes are quite different from the theoretical predictions. Overall, these results confirm the validity of the hydrodynamic "screening" hypothesis and ratify the neglect of hydrodynamical stresses in quantifying the linear rheology of Brownian rod suspensions. PMID- 18397102 TI - Dye reorientation as a probe of stress-induced mobility in polymer glasses. AB - The reorientation of dye molecules can be used to monitor the segmental dynamics of a polymer melt. We utilize this technique to measure stress-induced mobility in a lightly cross-linked poly(methyl methacrylate) (PMMA) glass during tensile creep deformation. At 377 K (18 K below the glass transition temperature Tg), the mobility increased by a factor of 100 during deformation with a stress of 20 MPa. Generally, the mobility increased as the stress, strain, and strain rate increased. After removing the stress, we observed that the enhanced mobility slowly disappeared during strain recovery. At 377 K, when the stress is lower than 11 MPa, almost no mobility enhancement was observed. Once the stress crossed this threshold value, the mobility dramatically increased. PMID- 18397103 TI - Model study for large deformation of physical polymeric gels. AB - A model for large deformation of polymer gels with physical cross-linking is developed and shown to be in good agreement with experimental stress-strain curves which show strain hardening in intermediate strains followed by strain softening in large deformations near the yield strain. The model takes into account the coil-helix transition equilibrium and allows for the distribution of the end-to-end distance. The gel is considered to be formed by long flexible chains and crystalline zones acting as junctions of the chains. The number of segments contained in a flexible chain is variable due to the equilibrium between the two regions. As the end-to-end distance increases due to the deformation, more and more segments are reeled out from the junction zone. Finally, one end of the chain is librated from the junction and the chain becomes dangling. The appearance of dangling chains causes the strain softening because they cease to contribute to the elasticity. From the parameter dependence of the stress-strain relations, it was found that the yield behavior depends strongly on the distribution of end-to-end distance. The yield strain is approximately given by the ratio of the upper limit of the number of segments and the average end-to-end distance. The standard deviation of the end-to-end distance affects significantly the width of the peak in the stress-strain curve, thus determining the degree of strain softening. PMID- 18397104 TI - Role of defects in the nonmonotonic behavior of secondary relaxation of polypropylene glycols. AB - A nonmonotonic relaxation kinetic model [Ya. Ryabov et al., J. Phys. Chem. B 105, 1845 (2001)] is successfully applied to describe an intriguing slow down in the dielectric secondary gamma relaxation of polypropylene glycols (PPGs) with increasing temperature near the glass transition. The anomalous behavior is interpreted as a result of two simultaneous events: A thermal activation and a defect formation in the hydrogen bonded network formed by molecules of PPGs. This new insight into the molecular mechanism, which is responsible for the suggested sensitivity of the secondary process in PPGs to the glass transition phenomenon, is compared to our previous results obtained in terms of the minimal model for secondary relaxations. PMID- 18397105 TI - A conductivity study and calorimetric analysis of dried poly(sodium 4-styrene sulfonate)/poly(diallyldimethylammonium chloride) polyelectrolyte complexes. AB - Ionically cross-linked polyelectrolyte complexes (PECs) of anionic poly(sodium 4 styrene sulfonate) (PSS) and cationic poly(diallyldimethylammonium chloride) (PDADMAC), xPSS.(1-x)PDADMAC, with molar fractions x ranging from 0.30 to 0.70, were prepared and subsequently dried. The PEC samples were analyzed by differential scanning calorimetry, and the ionic conductivity sigmadc of the samples was measured as a function of temperature by means of impedance spectroscopy. The thermograms display an endothermic peak in the temperature range of 90-143 degrees C, which is attributed to a glass transition of the PEC. The glass transition temperature Tg has a symmetric x dependence with a minimum at x=0.50. The temperature dependence of sigmadcT is not affected by the glass transition. The ionic conductivity of the samples before drying is three orders of magnitude larger than sigmadc after drying; nevertheless, their activation enthalpies are identical. Arrhenius parameters obtained from the systematic study of several PEC compositions are discussed. The ionic conductivity of the PSS-rich samples is significantly higher than sigmadc of PDADMAC-rich samples. This implies a relatively high Na+ mobility as compared to Cl(-) mobility in PEC. In contrast to the symmetric x dependence of Tg, the conductivity of PEC increases and the activation enthalpy decreases with increasing x in the investigated composition range. A strong x dependence of sigmadc is observed for PSS-rich PEC, which is attributed to a significant variation in the mobility of the charge carriers. PMID- 18397106 TI - Effect of interaction shape on the condensed toroid of the semiflexible chain. AB - We investigate how different microscopic interactions between semiflexible chain segments can qualitatively alter the physical properties of the condensed toroid. We propose a general form of the Hamiltonian of the toroid and discuss its analytic properties. For different interactions, the theory predicts different scaling behaviors of the mean toroidal and cross sectional radii, rc and rcross, as functions of the contour length L: (rc,rcross) approximately Lnu(Nc), with nu=(1/5,2/5) for the van der Waals-type, nu=(-1/3,2/3) for the Coulomb-type, and nu=(-1,1) for the delta-function-type attractions in the asymptotic limit. For the toroids with finite winding number Nc=100-400, we find nu approximately 0 for the Yukawa interaction with screening parameter kappa=0.5-1.0 and nu=0.1-0.13 for the van der Waals-type interactions. These findings could provide a possible explanation for the experimentally well known observation nu approximately 0 of the condensed DNA toroids. Conformational transitions are also discussed. PMID- 18397107 TI - Renormalized charge in a two-dimensional model of colloidal suspension from hypernetted chain approach. AB - The renormalized charge of a simple two-dimensional model of colloidal suspension was determined by solving the hypernetted chain approximation and Ornstein Zernike equations. At the infinite dilution limit, the asymptotic behavior of the correlation functions is used to define the effective interactions between the components of the system and these effective interactions were compared to those derived from the Poisson-Boltzmann theory. The results we obtained show that, in contrast to the mean-field theory, the renormalized charge does not saturate, but exhibits a maximum value and then decays monotonically as the bare charge increases. The results also suggest that beyond the counterion layer near to the macroion surface, the ionic cloud is not a diffuse layer which can be handled by means of the linearized theory, as the two-state model claims, but a more complex structure is settled by the correlations between microions. PMID- 18397108 TI - The initial single yeast cell adhesion on glass via optical trapping and Derjaguin-Landau-Verwey-Overbeek predictions. AB - We used an optical tweezer to investigate the adhesion of yeast Saccharomyces cerevisiae onto a glass substrate at the initial contact. Micromanipulation of free-living objects with single-beam gradient optical trap enabled to highlight mechanisms involved in this initial contact. As a function of the ionic strength and with a displacement parallel to the glass surface, the yeast adheres following different successive ways: (i) Slipping and rolling at 1.5 mM NaCl, (ii) slipping, rolling, and sticking at 15 mM NaCl, and (iii) only sticking at 150 mM. These observations were numerous and reproducible. A kinetic evolution of these adhesion phenomena during yeast movement was clearly established. The nature, range, and relative intensity of forces involved in these different adhesion mechanisms have been worked out as a quantitative analysis from Derjaguin-Landau-Verwey-Overbeek (DLVO) and extended DLVO theories. Calculations show that the adhesion mechanisms observed and their affinity with ionic strength were mainly governed by the Lifshitz-van der Waals interaction forces and the electrical double-layer repulsion to which are added specific contact forces linked to "sticky" glycoprotein secretion, considered to be the main forces capable of overcoming the short-range Lewis acid-base repulsions. PMID- 18397109 TI - Ultrafast exciton transfers in DNA and its nonlinear optical spectroscopy. AB - We have calculated the nonlinear response function of a DNA duplex helix including the contributions from the exciton population and coherence transfers by developing an appropriate exciton theory as well as by utilizing a projector operator technique. As a representative example of DNA double helices, the B-form (dA)10-(dT)10 is considered in detail. The Green functions of the exciton population and coherence transfer processes were obtained by developing the DNA exciton Hamiltonian. This enables us to study the dynamic properties of the solvent relaxation and exciton transfers. The spectral density describing the DNA base-solvent interactions was obtained by adjusting the solvent reorganization energy to reproduce the absorption and steady-state fluorescence spectra. The time-dependent fluorescence shift of the model DNA system is found to be ultrafast and it is largely determined by the exciton population transfer processes. It is further shown that the nonlinear optical spectroscopic techniques such as photon echo peak shift and two-dimensional photon echo can provide important information on the exciton dynamics of the DNA double helix. We have found that the exciton-exciton coherence transfer plays critical roles in the peculiar energy transfer and ultrafast memory loss of the initially created excitonic state in the DNA duplex helix. PMID- 18397110 TI - Gel-like behavior in aggrecan assemblies. AB - Aggrecan, a large biological polyelectrolyte molecule with a bottlebrush shape, forms complexes with hyaluronic acid (HA) that provide compressive resistance in cartilage. In solutions of aggrecan alone, the concentration dependence of the osmotic pressure Pi is marked by self-assembly of the molecules into aggregates. When HA is added to the solution at low aggrecan concentration c, the osmotic pressure is reduced, but in the physiological concentration range this trend is reversed. The osmotic modulus c partial differentialPi partial differentialc, which determines load bearing resistance, is enhanced in the HA-containing solutions. Dynamic light scattering (DLS) measurements show that the aggregates behave like microgels and that they become denser as the aggrecan concentration increases. The degree of densification is greatest at large distance scales in the microgels, but decreases at short distance scales. Measurements at higher resolution, involving small angle neutron scattering and small angle x-ray scattering (SAXS), confirm that at length scales shorter than 1000 angstroms, the density is independent of the concentration and that the individual bottlebrushes in the microgels retain their identity. The absence of collective diffusion modes in the relaxation spectrum, measured by DLS and neutron spin echo, corroborates the lack of interpenetration among the aggrecan subunits in the microgel. Complexation with HA modifies the long-range spatial organization of the microgels. Comparison of the scattering pattern of the individual aggrecan molecules obtained from SAXS measurements with that of the complexes measured by DLS shows that the aggrecan-HA structure is denser and is more uniform than the random microgels. This enhanced space-filling property allows higher packing densities to be attained, thus, optimizing resistance to osmotic compression. PMID- 18397111 TI - The influence of ion binding and ion specific potentials on the double layer pressure between charged bilayers at low salt concentrations. AB - Measurements of surface forces between double-chained cationic bilayers adsorbed onto molecularly smooth mica surfaces across different millimolar salt solutions have revealed a large degree of ion specificity [Pashley et al., J. Phys. Chem. 90, 1637 (1986)]. This has been interpreted in terms of highly specific anion binding to the adsorbed bilayers. We show here that inclusion in the double layer theory of nonspecific ion binding and ion specific nonelectrostatic potentials acting between ions and the two surfaces can account for the phenomenon. It also gives the right Hofmeister series for the double layer pressure. PMID- 18397112 TI - An entropy-based approach to the freezing of the generalized exponential model. PMID- 18397113 TI - Efficient algorithms for rigid body integration using optimized splitting methods and exact free rotational motion. PMID- 18397116 TI - Determination of critical network interactions: an augmented Boolean pseudo dynamics approach. AB - Network theory has established that highly connected nodes in regulatory networks (hubs) show a strong correlation with criticality in network function. Although topological analysis is fully capable of identifying network hubs, it does not provide an objective method for ranking the importance of a particular node by relating its contribution to the overall network response. Towards this end, the authors have developed an augmented Boolean pseudo-dynamics approach to a priori determine the critical network interactions in biological interaction networks. The approach utilises network topology and dynamic state information to determine the set of active pathways. The active pathways are used in conjunction with the key cellular properties of efficiency and robustness, to rank the network interactions based on their importance in the sustenance of network function. To demonstrate the utility of the approach, the authors consider the well characterised guard cell signalling network in plant cells. An integrated analysis of the network revealed the critical mechanisms resulting in stomata closure in the presence and absence of abscisic acid, in excellent agreement with published results. PMID- 18397117 TI - Control analysis of the role of triosephosphate isomerase in glucose metabolism in Lactococcus lactis. AB - Triosephosphate isomerase (TPI), which catalyses the conversion of dihydroxyacetone phosphate (DHAP) to glyceraldehyde-3-phosphate (G3P), was studied for its control on glycolysis and mixed acid production in L. lactis subspecies lactis IL1403 and L. lactis subspecies cremoris MG1363. Strains in which the TPI activity was modulated from 3%-225% (IL1403) or 13%-103% (MG1363) of the wild-type level were constructed by changing the expression of the tpiA gene. The enzyme was found to be present in high excess in the wild-type cells and 10% TPI activity still supported more than 70% of the wild-type glycolytic flux in both strains. Homolactic product formation was preserved throughout the range of TPI activities studied, although a slight increase in the amount of acetate and formate production was observed in the strains with strongly reduced TPI activity for both IL1403 and MG1363. The upstream metabolites glucose-6 phosphate, fructose-1,6-bisphosphate and DHAP in the IL1403 derivatives were essentially unchanged for TPI activities from 26% to 225%. At a TPI activity of 3%, the level of DHAP increased four times. The finding that an increased level of DHAP coincides with an increase in formate production is surprising and indicates that pyruvate formate lyase is not inhibited by DHAP under these conditions. PMID- 18397118 TI - Modelling biological modularity with CellML. AB - In recent years advances in the construction of mathematical models of biological systems have yielded an array of valuable constructs. The authors seek to provide a 'leading practice' method for implementing modularised kinetic mass-action models in order to obtain a number of advantages in model construction, validation and derived insights. The authors advocate the consideration of 'accounting cycles' or 'chains' to define 'functional' components and the separate consideration of 'messenger' components for mobile or diffusive molecular species. From a conceptual modularisation the authors illustrate, with an example drawn from signal transduction, a component-based formulation in the model exchange format cellular modelling markup language (CellML) 1.1 - demonstrating loose coupling between functionally-focused reusable components. Finally, the authors discuss the dilemmas associated with modelling protein-to protein interactions, and the vision for using future CellML enhancements to resolve potential duplications when combining independently developed models. PMID- 18397119 TI - Multistability of signal transduction motifs. AB - Protein domains are the basic units of signalling processes. The mechanisms they are involved in usually follow recurring patterns, such as phosphorylation/dephosphorylation cycles. A set of common motifs was defined and their dynamic models were analysed with respect to number and stability of steady states. In a first step, Feinberg's chemical reaction network theory was used to determine whether a motif can show multistationarity or not. The analysis revealed that, apart from double-step activation motifs including a distributive mechanism, only those motifs involving an autocatalytic reaction can show multistationarity. To further characterise these motifs, a large number of randomly chosen parameter sets leading to bistability was generated, followed by a bifurcation analysis of each parameter set and a statistical evaluation of the results. The statistical results can be used to explore robustness against noise, pointing to the observation that multistationarity at the single-motif level may not be a robust property; the range of protein concentrations compatible with multistationarity is fairly narrow. Furthermore, experimental evidence suggests that protein concentrations vary substantially between cells. Considering a motif designed to be a bistable switch, this implies that fluctuation of protein concentrations between cells would prevent a significant proportion of motifs from acting as a switch. The authors consider this to be a first step towards a catalogue of fully characterised signalling modules. PMID- 18397120 TI - Modelling the influence of activation-induced apoptosis of CD4+ and CD8+ T-cells on the immune system response of a HIV-infected patient. AB - On the basis of the human immunodeficiency virus (HIV) infection dynamics model proposed by Adams, the authors propose an extended model that aims at incorporating the influence of activation-induced apoptosis of CD4+ and CD8+ T cells on the immune system response of HIV-infected patients. Through this model, the authors study the influence of this phenomenon on the time evolution of specific cell populations such as plasma concentrations of HIV copies, or blood concentrations of CD4+ and CD8+ T-cells. In particular, this study shows that depending on its intensity, the apoptosis phenomenon can either favour or mitigate the long-term evolution of the HIV infection. PMID- 18397121 TI - Development of Free Neuromasts with Special Reference to Sensory Polarity in Larvae of the Willow Shiner, Gnathopogon elongatus caerulescens (Teleostei, Cyprinidae). AB - To find how larval fish sense mechanical stimuli via their free neuromasts, we examined morphological changes in free neuromasts in the larval willow shiner, Gnathopogon elongatus caerulescens. Free neuromasts were found on the body surface of newly hatched larvae and their number increased on both the head and trunk with larval growth. The apical surface of free neuromasts changed in outline from a circle to a lozenge shape as the number of sensory cells increased in the prelarval stage, and then the cupulae of the free neuromasts changed from a stick-like to a blade-like shape. Seven-day-old larvae were at the postlarval stage and had many free neuromasts that were nearly mature. All free neuromasts contained sensory cells of opposing polarity. The orientation of the maximum sensitivity of free neuromasts, decided from the polarity of the sensory cells, coincided with the minor axis of the lozenge-shaped outline of the apical surface of the free neuromasts, and was in the same axis as the direction in which the blade-like cupulae bent. The change to a blade-like shape would cause a stimulus parallel to the minor axis to be perceived as being stronger than the same stimulus from other directions. The polarity of trunk neuromasts was usually oriented along the antero-posterior axis of the fish body, but a few had dorso ventral orientation. On the head, free neuromasts were oriented on lines tangential to concentric circles around the eye. PMID- 18397122 TI - Correlation of the Period Length of Circadian Rhythms with the Length of Immaturity in Paramecium bursaria. AB - The circadian photoaccumulation rhythm of thirty strains of Paramecium bursaria collected at different places in Japan and China were measured with a microcomputer assisted data collection apparatus. Although most strains showed a period of 23-26 hours in LL, we found two strains of conspicuously different periods; a short period strain (UK1, 21.8 hr) and a long period strain (T316, 28.7 hr). F1 progeny from a cross between the short and the long period strains showed an intermediate period of about 24.7 hours (range 22.5-25.8 hr). The character was not distributed in a Mendelian ratio among the F1 progeny. We isolated a mutant (E2) with short period (21.8 hr) from the stock strain Kz1 by treatment with nitrosoguanidine (MNNG). The progeny of crosses between E2 and UK1, and between E2 and T316 exhibited the short period and the normal period phenotype respectively. Moreover, the progeny from a cross between E2 and a wild type strain (Sj2w) became sexually mature about 25 fissions after conjugation. This length of immaturity is much shorter than that of the progeny from wild type strains (about 50 fissions). This early maturation character was inherited to progeny in a Mendelian ratio. Homozygotes for the early maturation allele (EM2) exhibited mating ability about 15 fissions after conjugation. These data suggest that there is a correlation between the period length of the circadian rhythm and the length of immaturity after conjugation in Paramecium bursaria. PMID- 18397123 TI - Genetic divergence and evolutionary relationships of the old and new world emberizidae. AB - Genetic differentiation and evolutionary relationships were surveyed on 12 species of the Old and seven species of the New World Emberizidae by allozyme electrophoresis of 20 loci. Genetic variability of the Emberizidae is similar to those of the other Passeriformes. The degree of genetic differentiation in the family were large among species of the genus Emberiza of the Old World, and also among genera of the New World Emberizidae. Evolutionary relationships of the Emberizidae based on these genetic distances differed considerably from those of previous publications on some points: (1) Emberiza tristrami, E. elegans, E. bruniceps and E. schoeniclus were genetically much diverged from the other Emberiza as a species of the same genus. (2) Some genetic distances between Emberiza were larger than distances between subfamilies of the New World Emberizidae. (3) Species of the Cardinarinae examined genetically, belonged to the Emberizinae of the New World. Genetic data did not support the current classification that the Old World buntings arose from the New World forms by recent colonization. Discussion was made on the evolution of the Old and New World Emberizidae from the genetic view point. PMID- 18397124 TI - Histological Changes in the Pituitary, Thyroid Gland and Gonads of the Fourspine Sculpin (Cottus kazika) during Downstream Migration. AB - The Fourspine sculpin (Cottus kazika) is a catadromous fish which is widely distributed in the rivers of Japan. The fish was used to examine the relationship between the migration behavior and hormonal control by studying the histological changes in the pituitary gland, thyroid gland and gonads during its downstream migration. By use of the immonocytochemical and histochemical techniques, 7 types of cells were identified in the pituitary gland namely; immunoreactive (ir)-PRL, GH, TSH, GTH, ACTH, MSH and SL cells. From among the first 4 types of the aforementioned cells, remarkable histological changes were observed in cells containing ir-GTH during the downstream migration. At this time also, the gonads were obsereved to be well developed, while the thyroid glands did not show clear changes morphologically. These results suggest that the gonadotropin regulates gonadal development in the Fourspine sculpin during downstream migration and possibly sex hormones synthesized by the gonads cause the downstream migration of this catadromous fish. PMID- 18397125 TI - A continuum of service: integration at Baylor Health Care System. PMID- 18397127 TI - Finding the "constructive" in criticism. PMID- 18397128 TI - Improving equipment purchasing decisions through human factors engineering. PMID- 18397129 TI - The word for quick is wiki-wiki. PMID- 18397130 TI - Infection control practices for biomeds. PMID- 18397131 TI - Video carts: a logical set-up equals simple troubleshooting. PMID- 18397132 TI - Competency assessments help strengthen promotion system, develop staff skills. PMID- 18397133 TI - Portable power devices. PMID- 18397134 TI - An inside look: how certification test questions make the grade. PMID- 18397135 TI - Product quality assurance testing: one hospital's experience. PMID- 18397136 TI - 2008 National Patient Safety Goals... communication is key to infection control with equipment. PMID- 18397137 TI - Clinical alarm management: a team effort. PMID- 18397138 TI - Who's monitoring your lab? Choosing the right facility monitoring system. PMID- 18397139 TI - AAMI releases new guidance for software validation. PMID- 18397140 TI - The microbial world and fluids in dialysis. AB - The fluids used in dialysis are all water based. Water, which is necessary for life, is also a good environment for micro-organisms. The result of this is quite simply that microbial growth, i.e., increased numbers of micro-organisms, results in the presence of endotoxins and the formation of metabolites. The situation is not favorable for dialysis and needs to be understood and corrected. In order to act in the right way we must analyze and synthesize the available information, which will lead us to decide what actions and precautions are necessary. The possible negative effects of a too-high microbiological content in dialysis fluid and the importance of hygiene have been well-documented. This chain of events depends on ensuring that the information and thus the understanding we get from that information is accurate. If it is not accurate, any actions taken may be inadequate and result in a situation we can no longer control. The patient will be in contact with the dialysis fluid in every session of dialysis due to the phenomenon of backfiltration, which means that anywhere from 100 ml to multiple liters of dialysis fluid is filtered over the dialyzer membrane and into the blood. The problems that may occur when contaminated dialysis fluid is used range from the acute pyrogenic reaction to chronic reactions over time where no acute symptoms are identified. The immune defense system is, however, constantly tested by the presence of body foreign components in the dialysis fluid. This paper will discuss the microflora (the micro-organisms present in the microbial community) that occur in systems of fluids in dialysis, limits, and methods of cultivation and disinfection. Results presented are original data examples out of some 350 investigations of fluid systems in dialysis units around the world, using analytic methods. PMID- 18397141 TI - A call for change. PMID- 18397143 TI - Inside the numbers... flex time grows in popularity. PMID- 18397144 TI - FDA safety alert: focus on communication. PMID- 18397145 TI - Digital cameras. PMID- 18397154 TI - The changing face of education. PMID- 18397155 TI - Anaplastic large cell neuroblastoma. AB - Anaplastic large cell neuroblastomas (ALCNB) are a subset of undifferentiated neuroblastomas with marked pleomorphic and anaplastic features that render them diagnostically challenging. We reviewed the records of all patients diagnosed with ALCNB at Children's Healthcare of Atlanta (Egleston Children's Hospital) for their clinical, biologic, and pathologic characteristics and their treatment outcomes. From 1998 to 2006, 7 patients were diagnosed with ALCNB. All patients presented with abdominal-pelvic masses, 3 of them of adrenal origin and 2 with thoracic extension, with clinical stages 3 or 4, and were considered to have high risk disease. The N-MYC oncogene was amplified in 3 cases and catecholamines were elevated in 5 of 6 patients tested. All pretreatment tumors demonstrate pleomorphic, anaplastic morphology with bizarre mitoses admixed with undifferentiated but monomorphic cells with minimal if any neuropil or neuro ganglionic differentiation. Immunohistochemical markers for neuron specific enolase (NSE) and synaptophysin were strongly positive in all specimens and chromogranin in 4 of 5. Interestingly, all tumors showed strong Fli-1 nuclear positivity despite a negative CD-99 stain. However, reverse transcription polymerase chain reaction or fluorescent in-situ hybridization testing for Ewing sarcoma transcripts was negative in 4 available specimens. This same Fli-1 antibody had tested negative on 30 conventional neuroblastomas, indicating a peculiar cross reactivity with this subset of ALCNB. Posttreatment biopsies showed maturation changes to more conventional neuroblastoma histology in 5 of the 7 cases. Follow-up ranged from 9 months to 4 years from diagnosis (median: 25 months). Five patients are still alive after treatment, 1 died 9 months after diagnosis, and another patient refused high-risk therapy and progressed and died 9 months from diagnosis. Anaplastic large cell neuroblastomas are a subset of undifferentiated neuroblastomas characterized by the absence or marked paucity of histologic clues for the diagnosis of neuroblastoma. Although all these tumors are strongly positive for NSE and synaptophysin, they also show Fli-1 positivity. However, they are negative by molecular testing for EWS transcripts, and they are immunohistochemically negative for CD99. The true neuroblastic nature of these tumors is supported by the N-MYC oncogene amplification in some of them, catecholamine production, immunohistological reactivity, and their posttherapy maturation to a more recognizable neuroblastic morphology. Although follow-up is still somewhat limited, the response and survival of the patients in our institution is better than a previous European study that indicated an aggressive clinical behavior of these tumors, although treatment modalities were not described in that report. Further study of this variant of neuroblastoma with more patients is required to determine optimal therapy, more accurately predict outcome, and to ascertain if ALCNB are a distinct biologic group of neuroblastomas. PMID- 18397156 TI - Multipotent adult progenitor cells acquire periodontal ligament characteristics in vivo. AB - Mesenchymal stem cells (MSC) and multipotent adult progenitor cells (MAPC) copurify from the bone marrow. Past studies have shown that bone marrow-derived cells transplanted into the periodontium obtain characteristics of several periodontal cells types, such as alveolar bone, periodontal fibroblasts, and cementoblasts. Bone marrow-derived cell populations are heterogeneous mixtures of different cell types. Characterization of each individual cell type in the bone marrow has not been completed. In this study, the MAPC were isolated from the bone marrow of 3-week-old rats by selecting attached cells that did not express leukocyte (CD45), erythroid (HIS49), or the MSC marker CD44. DiI-labeled male MAPC were injected into the periodontal ligament (PDL) of female adult rats. After 3 weeks, the morphology of the DiI-labeled cells was determined. Periodontal tissues sections that included alveolar bone, PDL, cementum, and dentin were also immunostained for collagen I and III after 3 days, 3 weeks, and 6 weeks following injection of male MAPC into the female PDL. Male and female cells were differentiated by fluorescent in situ hybridization tagging of the X and Y chromosomes. Our results show that DiI-labeled MAPC were spherical in shape before injection but after 3 weeks a portion of the injected MAPC obtained the characteristic spindle shape of PDL cells with an orientation parallel to the surrounding PDL cells. Consistent with the DiI results, male MAPC had a spherical shape on immunostained sections 3 days after injection and expressed collagen I and III at significantly reduced levels as compared with the endogenous female PDL. Six weeks after injection, the MAPC expressed both collagen I and collagen III at levels similar to the endogenous surrounding PDL and obtain a morphology characteristic of PDL. In conclusion, MAPC obtain PDL-like characteristics over a 6-week period after injection into the PDL. PMID- 18397157 TI - Robotic versus standard laparoscopic partial/wedge nephrectomy: a comparison of intraoperative and perioperative results from a single institution. AB - PURPOSE: Laparoscopic partial/wedge nephrectomy, similar to laparoscopic radical prostatectomy, is a technically challenging procedure that is performed by a limited number of expert laparoscopic surgeons. The incorporation of a robotic surgical interface has dramatically increased the use of minimally invasive pelvic surgery such that robotic laparoscopic radical prostatectomy is commonly performed even by laparoscopically naive surgeons. This analysis compares the outcomes of our initial experience with robot-assisted laparoscopic partial nephrectomy (RLPN) performed by an experienced open surgeon to that of standard laparoscopic partial nephrectomy (LPN) performed by two experienced laparoscopic surgeons. PATIENTS AND METHODS: We reviewed the medical records of 11 consecutive patients who underwent 12 standard LPNs (EMM, RVC) (one patient had two unilateral tumors) and 10 consecutive patients (representing the first 11 of such robotic procedures performed at our institution) who underwent 11 RLPNs (one patient had bilateral tumors managed in an asynchronous manner) (DKO). RESULTS: The mean tumor size was 2.3 cm (range 1.7-6.2 cm) for LPN and 3.1 cm (range 2.5-4 cm) for RLPN. The mean total procedure time was 289.5 minutes (range 145-369 min) for LPN and 228.7 minutes (range 98-375 min) for RLPN (P=0.102). The mean estimated blood loss was 198 mL (range 75-500 mL) for LPN v 115 mL (25-300 mL) for RLPN (P=0.169). The mean warm ischemia time was 35.3 minutes (range 15-49 min) in the LPN group and 32.1 minutes (range 30-45 minutes) in the RLPN group (P=0.501). CONCLUSIONS: Introducing a robotic interface for laparoscopic partial/wedge resection allowed a fellowship-trained urologic oncologist with limited reconstructive laparoscopic experience to achieve results comparable to those for laparoscopic partial/wedge resection performed by experienced laparoscopic surgeons. In this regard, the learning curve appears truncated, similar to that with robot-assisted laparoscopic prostatectomy. PMID- 18397161 TI - Time domain reflectometry. PMID- 18397159 TI - Bilateral thyroid hematomas after fine-needle aspiration causing acute airway obstruction. AB - BACKGROUND: Fine-needle aspiration of the thyroid gland is a common procedure that is increasingly being used to evaluate thyroid nodules incidentally found by other imaging means such as computed tomography scans. Rare complications include hematoma, transient vocal cord paralysis, seeding of carcinoma cells, and infection. Of these, hematoma is the most common and is usually small and adequately treated with compression of the site. Only two cases have been previously reported describing large thyroid hematoma after fine-needle aspiration resulting in acute airway compromise and requiring surgical decompression. SUMMARY: We report an interesting case of large bilateral thyroid hematomas after routine fine-needle aspiration causing acute airway obstruction in a patient with hypertension and end stage renal disease taking aspirin. PMID- 18397162 TI - 50 years of physical chemistry, a personal account. PMID- 18397164 TI - When polymer science looked easy. PMID- 18397163 TI - Of physical chemistry and other activities. PMID- 18397165 TI - Polymer conformation and dynamics in solution. PMID- 18397166 TI - Laser control of molecular processes. PMID- 18397167 TI - Congeners of the enediyne neocarzinostatin chromophore: designed agents for bulged nucleic acid targets. AB - Of the commonly recognized structural elements within nucleic acids, bulges are among the least developed as targets for small molecules. Bulges in DNA and RNA have been linked to biomolecular processes involved in numerous diseases, thus probes with affinity for these targets would be of considerable utility to chemical biologists and medicinal chemists. Despite such opportunity, there is a dearth of small molecules available with affinity for bulges, which has hampered exploitation of these key targets. We have used guided chemical synthesis to prepare small molecules capable of binding to DNA and RNA bulges. Our design is based on a template which mimics a metabolite of the enediyne neocarzinostatin. The key spirocylic building block was formed through an intramolecular aldol process and the parent template shows pronounced affinity for 2 base bulges. Functionalization with specific aminosugar moieties confers nanomolar binding affinity for selected bulged DNA targets, and installation of reactive functional groups allows covalent modification of bulges. These rationally designed agents can now be used to study the stereochemistry and architecture of bulge-drug complexes and investigate the molecular biology of bulge induced processes. Members of this class have been shown to induce slipped synthesis of DNA, suggesting the agents, in addition to recognizing and binding to pre-formed bulges, can also induce bulge formation on demand. PMID- 18397168 TI - Biosynthesis of enediyne antitumor antibiotics. AB - The enediyne polyketides are secondary metabolites isolated from a variety of Actinomycetes. All members share very potent anticancer and antibiotic activity, and prospects for the clinical application of the enediynes has been validated with the recent marketing of two enediyne derivatives as anticancer agents. The biosynthesis of these compounds is of interest because of the numerous structural features that are unique to the enediyne family. The gene cluster for five enediynes has now been cloned and sequenced, providing the foundation to understand natures' means to biosynthesize such complex, exotic molecules. Presented here is a review of the current progress in delineating the biosynthesis of the enediynes with an emphasis on the model enediyne, C-1027. PMID- 18397169 TI - Towards photoswitchable enediyne antibiotics: single and two-photon triggering of bergman cyclization. AB - The concept of photoswitchable enediynes, which are stable in the dark but undergo efficient cycloaro-matization reaction to produce p-benzyne diradical after irradiation with light of an appropriate wavelength, is discussed. Two novel methods for the generation of reactive enediyne compounds from thermally stable precursors have been developed. In the first approach, one of the triple bonds of cyclodeca-3-ene-1,5-diynes is replaced with cyclopropenone group. Cyclopropenone-containing enediyne precursors are unable to undergo cycloaromatization because the enediyne fragment is incomplete. Photolysis of cyclopropenones results in the efficient decarbonylation and the regeneration of a triple bond thus completing the enediyne pi-system. The second method employs photo-Wolff reaction to achieve ring contraction of stable eleven-membered ring precursor enediynes. Benzannulated cyclic enediynes produced by the photodecomposition of enediyne precursor containing 2-diazo-1,3-diketones possess enolized beta-ketoester fragment and undergo remarkably facile tau(36)o = 5 min - 3 h) Bergman cyclization. The generation of reactive enediyne was also achieved with NIR light by non resonant two-photon excitation. PMID- 18397170 TI - The critical distance for the cycloaromatization reactions of enediynes. AB - It has been shown that the enediyne anticancer antibiotics e.g., calicheamicin, neocarzinostatin and others cleave DNA via the putative intermediate 1,4 diradical formed in the Bergmann cyclization and are thus useful for cancer chemotherapy. The pharmacological activity of these drugs is based, in general, on the activation of the pharmacophore and the subsequent cyclization leading to the formation of a radical, the rate of which is, in part, based on the terminal distance of the 1,5-diyne functionality, also known as the critical distance. But the critical distance alone cannot govern the rate of cyclization of the enediynes. A theoretical model has been developed to predict the rate of cyclization, and the thus the pharmacological activity, of these antibiotics based on the critical distance and the energy of activation. PMID- 18397171 TI - Synthesis of highly strained enediynes and dienediynes. AB - Enediynes continue to fascinate scientists working in various domains because of their structural complexity and fascinating biological mode of action. They represent a masterpiece of nature's ingenuity. Besides the warhead which is the enediyne moiety, these molecules are equipped with a locking device, a delivery system and a chemical trigger for activation. Upon triggering, the molecules become active and undergo a thermal rearrangement that was disclosed in the early '70 by Masamune and Bergman and commonly known as Bergman cyclization. The reaction is believed to precede through a diradical benzenoid species (a p benzyne). This review describes the various strategies employed for the synthesis of highly strained enediynes and dienediynes, both naturally occurring and the designed ones. PMID- 18397172 TI - Small molecule antagonists of the corticotropin releasing factor (CRF) receptor: recent medicinal chemistry developments. AB - Antagonists of the corticotropin releasing factor (CRF or CRH) receptor have shown promise for the treatment of anxiety, depression, and irritable bowel syndrome. In the present article, medicinal chemistry developments surrounding small molecule CRF receptor antagonists are reviewed, focusing on publications and patents from mid-2004 through the first quarter of 2006. While the CRF type 2 receptor remains an intractable target, incremental progress has been made in the search for drug-like antagonists of the CRF type 1 receptor. Most recent work has not ventured far from previously-established pharmacophoric topologies. A common theme in recent patent disclosures is the addition of novel polar substituents to known heterocyclic core structures to reduce overall lipophilicity. New disclosures of pharmacokinetic (PK) data for several series of antagonists reveal that achieving appropriate PK remains a challenge for the field. The recent publication of selection patents and patents relating to salt and crystal forms of particular compounds suggests that several second generation compounds are nearing or have entered clinical development. PMID- 18397173 TI - Recent progress in the discovery of novel glucocorticoid receptor modulators. AB - Glucocorticoids have been used in modern clinical practice for over fifty years. Although they have demonstrated potent anti-inflammatory and immunosuppressive activities, their association with debilitating and life-threatening side effects has been a major drawback. Recent insights into glucocorticoid biology have lent support to the hypothesis that the glucocorticoid anti-inflammatory activities could be dissociated from their adverse side effects. Inspired by these biological findings, the search for dissociated glucocorticoid receptor agonists has intensified. Antag-onists of the glucocorticoid receptor that offer therapeutic benefits for the treatment of diseases such as diabetes have also been pursued. These efforts have been partly focused on the development of tissue, especially liver, selective glucocor-ticoid receptor antagonists, which are thought to have improved safety profiles. This review offers a summary of the research and development activities in this field and covers journal and patent publications from 2003 to March 2006. PMID- 18397174 TI - Enediynes and related structures in medicinal and biorganic chemistry. PMID- 18397175 TI - HPA axis. PMID- 18397176 TI - VLDL exits from the endoplasmic reticulum in a specialized vesicle, the VLDL transport vesicle, in rat primary hepatocytes. AB - The movement of VLDL [very-LDL (low-density lipoprotein)] from the ER (endoplasmic reticulum) to the Golgi is required for its eventual secretion from hepatocytes and represents a potential target in controlling elevated concentrations of its metabolite LDL, the major determinant of atherosclerosis. To study this process, an in vitro ER-budding assay was developed to examine the generation of the VTV (VLDL transport vesicle) and PTV (protein transport vesicles) using ER isolated from [(14)C]TAG (triacylglycerol) and [(3)H]protein labelled primary rat hepatocytes. VTVs do not contain albumin, as determined by immunoblots. VTVs were distributed in light-density fractions, whereas PTVs were mainly in the mid-portion of the sucrose gradient. Electron microscopy revealed that VTVs were larger ( approximately 100-120 nm) in size than PTVs ( approximately 55-70 nm). ER from 0.4 mM OA (oleic acid)-treated hepatocytes budded VTVs of a lighter density as compared with VTVs budded from ER of 0.1 mM or 0.004 mM OA-treated hepatocytes. The generation of VTVs from rat hepatic ER required cytosol, ATP, Sar1 (a GTPase) and incubation at 37 degrees C. Proteinase K treatment did not degrade the VTV cargo protein, apoB100 (apolipoprotein 100), indicating that VTVs were sealed. Immunoblots showed that VTV concentrated apoB100, Sar1 and rSec22b, and excluded albumin and calnexin. VTVs were shown to fuse with cis-Golgi and delivered their cargo to the Golgi lumen, as determined by in vitro fusion, and acquired endoglycosidase H resistance. These results suggest that a new ER-derived transport vesicle (VTV) has been identified and characterized which transports nascent VLDL from the hepatic ER to the Golgi. PMID- 18397177 TI - Endosomal NADPH oxidase regulates c-Src activation following hypoxia/reoxygenation injury. AB - c-Src has been shown to activate NF-kappaB (nuclear factor kappaB) following H/R (hypoxia/reoxygenation) by acting as a redox-dependent IkappaBalpha (inhibitory kappaB) tyrosine kinase. In the present study, we have investigated the redox dependent mechanism of c-Src activation following H/R injury and found that ROS (reactive oxygen species) generated by endosomal Noxs (NADPH oxidases) are critical for this process. Endocytosis following H/R was required for the activation of endosomal Noxs, c-Src activation, and the ability of c-Src to tyrosine-phosphorylate IkappaBalpha. Quenching intra-endosomal ROS during reoxygenation inhibited c-Src activation without affecting c-Src recruitment from the plasma membrane to endosomes. However, siRNA (small interfering RNA)-mediated knockdown of Rac1 prevented c-Src recruitment into the endosomal compartment following H/R. Given that Rac1 is a known activator of Nox1 and Nox2, we investigated whether these two proteins were required for c-Src activation in Nox deficient primary fibroblasts. Findings from these studies suggest that both Nox1 and Nox2 participate in the initial redox activation of c-Src following H/R. In summary, our results suggest that Rac1-dependent Noxs play a critical role in activating c-Src following H/R injury. This signalling pathway may be a useful therapeutic target for ischaemia/reperfusion-related diseases. PMID- 18397178 TI - Alemtuzumab induction and triple maintenance immunotherapy in kidney transplantation from donors after cardiac death. AB - We have used alemtuzumab in combination with triple maintenance immunosuppression in renal transplantation from donors after cardiac death between 2002 and 2006. We compared outcomes of induction therapy with alemtuzumab with interleukin-2 (IL 2) receptor antagonists (RA) and anti-lymphocyte antibodies. We used a retrospective sequential study design to examine 170 recipients of kidneys from donor after cardiac death (DCD) for survival, graft survival, time to first rejection, glomerular filtration and complications. Patients were stratified into high-risk and low-risk groups based on the following criteria: panel of reactive antibodies >20%, retransplants, Afro-American race. Induction with alemtuzumab was compared with anti-thymocyte globulin (ATG) in the high-risk and with IL-2RA in the low-risk group. Patients received triple immunosuppression with steroids, mycophenolate mofetil and calcineurin inhibitors. Patient survival, graft survival, rejection rate and glomerular filtration rate did not significantly differ between patients treated with alemtuzumab versus IL-2RAs or ATG. There was a trend towards reduced graft- and patient survival in the alemtuzumab group. There was an increased incidence of cytomegalovirus (CMV) infections in the alemtuzumab-induced group and a trend towards increased BK virus and bacterial infections. Induction of DCD kidney transplants with alemtuzumab compared to IL 2RA and ATG has no significant impact on acute rejection. It appears however that CMV infections are increased in patients induced with alemtuzumab. We therefore conclude that induction with alemtuzumab does not confer any advantage over traditional induction agents. PMID- 18397179 TI - Insect sperm motility. AB - The flagellosperm of insects, although following a general ground plan, exhibit considerable variation in morphology and ultrastructure across taxa, consistent with a history of rapid and divergent evolution. Sperm competition, which occurs when sperm of two or more males compete for the fertilization of a female's ova, has been recognized as a significant driving force in the evolution of insect sperm structure. Despite a considerable volume of data on sperm morphology, little is known about the motility of insect sperm. Understanding insect sperm motility would help to refine models of sexual selection on insect sperm, and would throw light on the selective mechanisms that shape insect sperm structure and function. This review updates our present knowledge of the proximate and ultimate aspects of insect sperm motility. PMID- 18397180 TI - Does size matter for hypoxia tolerance in fish? AB - Fish cover a large size range, from milligrams to tonnes, and many of them are regularly exposed to large variations in ambient oxygen levels. For more than half a century, there have been various, often divergent, claims regarding the effect of body size on hypoxia tolerance in fish. Here, we attempt to link old and new empirical data with the current understanding of the physiological mechanisms behind hypoxia tolerance. Three main conclusions are drawn: (1) body size per se has little or no impact on the ability to take up oxygen during hypoxic conditions, primarily because the respiratory surface area matches metabolic rate over a wide size range. If size-related differences are seen in the ability for oxygen uptake in a species, these are likely to reflect adaptation to different life-styles or habitat choice. (2) During severe hypoxia and anoxia, where fish have to rely on anaerobic ATP production (glycolysis) for survival, large individuals have a clear advantage over smaller ones, because small fish will run out of glycogen or reach lethal levels of anaerobic end products (lactate and H(+)) much faster due to their higher mass-specific metabolic rate. (3) Those fish species that have evolved extreme adaptations to hypoxia, including haemoglobins with exceptionally high oxygen affinities and an alternative anaerobic end-product (ethanol), reveal that natural selection can be a much more powerful determinant of hypoxia tolerance than scaling of physiological functions. PMID- 18397181 TI - Invasive Haemophilus influenzae disease in children less than 5 years of age in Manhica, a rural area of southern Mozambique. AB - OBJECTIVE: To measure the disease burden and epidemiological characteristics of invasive Haemophilus influenzae in rural Mozambican children. METHODS: As part of the clinical management of children admitted to Manhica District Hospital, blood and cerebrospinal fluid samples were collected between May 2001 and April 2005 for children aged <5 years. The level of antibiotic resistance of the isolates was analysed. RESULTS: During the surveillance period, there were 106 episodes of invasive H. influenzae disease. The estimated minimum incidence rate of invasive disease among children <5 years of age was 125/100,000 per child-year-at-risk. Fifty-six (59/106) per cent of cases were infants aged 3 to <12 months. Confirmed meningitis explained 16% of the episodes (n = 17) but was probably underestimated, as meningitis surveillance was not well implemented in the setting during the study. The case-fatality-rate was 21%, being highest among children with meningitis (odds ratio = 4.38, P = 0.011). Resistance to the antibiotics most commonly used in Mozambique was high--chloramphenicol 39%, ampicillin 35% and cotrimoxazol 74%-- and had increased over the years (P < 0.001 for chloramphenicol). CONCLUSION: Invasive H. influenzae disease is of considerable public health importance in Mozambique; implementing H. influenzae type b vaccination in sub-Saharan Africa has the potential to increase child survival. PMID- 18397182 TI - Noble goals, unforeseen consequences: control of tropical diseases in colonial Central Africa and the iatrogenic transmission of blood-borne viruses. AB - BACKGROUND: In southern Cameroon, 40-50% of individuals born before 1945 have antibodies against hepatitis C virus (HCV), suggesting massive iatrogenic transmission of at least one blood-borne virus in the region of the world where SIV(cpz) emerged into HIV-1. OBJECTIVE: To estimate the potential role of disease control programs that used intravenous (IV) drugs in the transmission of blood borne viruses, especially HCV. Methods We reviewed, for 1921-1959, records of health services in Cameroun, Oubangui-Chari, Gabon and Moyen-Congo. We calculated the incidence of diseases whose treatment required the administration of IV drugs, and compared these with previously published data on HCV prevalence. RESULTS: Several IV drugs were used against African trypanosomiasis, leprosy, yaws and syphilis. However, yaws was the only disease whose incidence was high enough so that up to half of some birth cohorts could have acquired HCV. Yaws incidence varied dramatically between regions, and was often >200 per 1000 per year in southern Cameroon, where extremely high HCV prevalence was found. Yaws incidence peaked between 1935 and 1955, a period which coincided with the emergence of HCV and HIV. CONCLUSION: Age, geographical and temporal distributions of yaws suggest that the HCV epidemic in Cameroon was driven by campaigns against yaws (and, secondarily, syphilis) using arsenicals and other metallic drugs. The same interventions may have exponentially amplified other blood-borne viruses, including SIV(cpz)/HIV-1. PMID- 18397183 TI - High-resolution 3D quantitative analysis of caveolar ultrastructure and caveola cytoskeleton interactions. AB - Caveolae are characteristic invaginations of the mammalian plasma membrane (PM) implicated in lipid regulation, signal transduction and endocytosis. We have employed electron microscope tomography (ET) to quantify caveolae structure function relationships in three-dimension (3D) at high resolution both in conventionally fixed and in fast-frozen/freeze-substituted (intact) cells as well as immunolabelled PM lawns. Our findings provide a detailed quantitative comparison of the average caveola dimensions for different cell types including tissue endothelial cells and cultured 3T3-L1 adipocytes. These studies revealed the presence of a spiked caveolar coat and a wide caveolar neck open to the extracellular milieu that is sensitive to conventional fixation; the neck region appeared to form a specialized microdomain with associated cytoplasmic material. In endothelial cells in situ in pancreatic islets of Langerhans, the diaphragm spanning the caveolar opening was clearly resolved by ET, and the involuted 3D topology of the cell surface mapped to measure the contribution of caveolar membranes to local increases in the surface area of the PM. The complexity of connections among caveolae and to the actin cytoskeleton and microtubules suggests that individual caveolae may be interconnected through a complex filamentous network to form a single functional unit. PMID- 18397184 TI - Antral mass due to cytomegalovirus infection requiring gastrectomy in a liver transplant recipient. AB - The incidence of cytomegalovirus (CMV) infection after liver transplantation (LT) has decreased in recent years. Advances in immunosuppression and CMV prophylaxis have improved the management of CMV disease. Organ involvement is infrequent and gastrointestinal CMV disease is quite rare. Few cases of an antral mass due to CMV infection have been described; those reported to date have mostly been in patients with acquired immunodeficiency syndrome. We describe a case of a CMV seronegative liver transplant patient who received a seropositive liver graft. Owing to gastrointestinal complaints, CMV prophylaxis was stopped one month after LT. The patient developed an antral mass due to CMV infection and an anastomotic biliary stricture. Antigenemia became negative with ganciclovir, but this treatment did not eliminate the mass. Ganciclovir resistance was ruled out as well as other causes of antral mass, especially malignancy. The patient finally required gastrectomy and hepaticojejunostomy. We conclude that CMV disease is less common today but should be included in the diagnosis of gastrointestinal mass after transplantation. PMID- 18397185 TI - Long-term outcome of benign fibroadenomas treated by ultrasound-guided percutaneous excision. AB - Surgical as well as conservative treatment has been described for fibroadenomas. Both have disadvantages. A minimally invasive treatment, ultrasound-guided, vacuum-assisted percutaneous excision has been shown to facilitate the removal of all imaged evidence of benign breast lesions, including fibroadenomas up to 3 cm in diameter. This study is performed to assess the long-term outcome of ultrasound-guided percutaneous excision as a minimally invasive treatment for fibroadenomas. A retrospective review of 69 consecutive fibroadenomas treated with ultrasound-guided percutaneous excision between May, 2001 and December, 2005 was carried out. All these lesions underwent percutaneous excision of all imaged lesion evidence. Clinical and sonographic follow-up was recommended for all patients every 6 months. Initial size, location, and patient age were recorded for each treated lesion. Of 69 lesions treated, 52 were available for follow-up. The median follow-up period was 22 months, with a range of 7 to 59 months. At 6 months, there were no fibroadenoma recurrences. Follow-up sonography demonstrated recurrences in 13 lesions distributed across eight patients. The overall recurrence rate was 15% (8/52) with an actuarial recurrence rate of 33% at 59 months. All of the recurrences were in lesions which were larger than 2 cm in size at initial presentation. Our data suggest that the mechanism of recurrence is the regrowth of retained lesion fragments too small to be detected by ultrasound--not the incomplete excision of all imaged lesion evidence. Despite successful percutaneous excision, fibroadenomas do recur. Lesions smaller than 2 cm in size, so treated, do not need additional therapy or surveillance. Fibroadenomas larger than 2 cm are prone to recurrence and require additional treatment. PMID- 18397186 TI - Interaction between seroreactivity to microbial antigens and genetics in Crohn's disease: is there a role for defensins? AB - Antibodies against different microbial epitopes are associated with disease phenotype, may be of diagnostic importance and may reflect a loss of tolerance in Crohn's disease (CD). Recently, an association was reported between the presence of these antibodies and mutations in pattern receptor genes. Our aim was to investigate whether mutations in various genes other than NOD2/CARD15 or TLR4 associated with CD (NOD1/CARD4, DLG5 and DEFB1) may influence the presence of antibodies against bacterial proteins and carbohydrates in a Hungarian cohort of CD patients. Three hundred and seventy-six well-characterized, unrelated, consecutive CD patients (male/female: 191/185, age at onset: 29.1 +/- 12.9 years, duration: 7.9 +/- 11.7 years) were investigated. Sera were assayed for anti-Omp, anti-Saccharomyces cerevisiae antibodies (ASCAs) immunoglobulin (Ig) A and IgG, and antibodies against a mannan epitope of S. cerevisiae (gASCA), laminaribioside (ALCA), chitobioside (ACCA), and mannobioside (AMCA). NOD1/CARD4, DLG5 and DEFB1 variants were tested by polymerase chain reaction-restriction fragment length polymorphism, and DEFB1 was genotyped in a subgroup of 160 patients. Detailed clinical phenotypes were determined by reviewing the patients' medical charts. The carriage of DEFB1 20A variant alleles less frequently led to antiglycan positivity compared with patients without (29.6% vs 46.2%, OR: 0.49, 95% CI: 0.25 0.97), regardless of disease location or behavior. Similar tendency was observed for DEFB1 44G (present: 21.6% vs absent: 10.2%, P = 0.06) and ALCA. A gene or serology dosage effect was not observed. However, no association was found between the DEFB1 G52A, DLG5 R30Q, and NOD1/CARD4 E266K variants and any of the serology markers. We found that variants in human beta-defensin 1 gene are inversely associated with antiglycan antibodies, further confirming an important role for innate immunity in the pathogenesis of CD. PMID- 18397187 TI - The presence of DRB1*01 allele in multiple myeloma patients is associated with an indolent disease. AB - The human leukocyte antigen (HLA) system could play an essential role in multiple myeloma (MM) disease control. This report describes the results comparing HLA DRB1 phenotypic frequencies in 181 MM patients (53 smoldering/indolent MM and 128 symptomatic MM patients) and healthy individuals. Higher DRB1*01 phenotypic frequencies were found in the smoldering patients compared with symptomatic MM patients (38% vs 14%, P = 0.001) and with the healthy individuals (38% vs 22%, P = 0.01). Additionally, higher DRB1*07 phenotypic frequencies were found in symptomatic MM compared with control population (38% vs 28%, P = 0.01). The present data suggest that HLA-DRB1*01 individuals may have a better ability to efficiently present myeloma-related antigens to immunocompetent cells, which could favor a better immune response against the tumor. This would translate into a more appropriate disease control associated with more indolent disease and prolonged survival. PMID- 18397188 TI - Two novel alleles HLA-B*9536 and B*4612 were identified in a healthy Chinese individual. AB - The novel HLA-B*9536 and B*4612 alleles were identified by sequence-based typing in China. PMID- 18397189 TI - Infertile boars with knobbed and immotile short-tail sperm defects in the Finnish Yorkshire breed. AB - In the period 1996-2006 two specific sperm defects, the knobbed acrosome (KA) defect and the immotile short-tail sperm (ISTS) defect, showed a strong negative association with fertility in Finnish breeding boars. In this study, we examined the incidence of these two sperm defects in two pig breeds, their effects on fertility and their associations with sperm morphology and testicular histology. Semen samples from 2048 (1097 Yorkshire, 951 Landrace) boars were collected. None of the Landrace boars revealed either the KA defect or the ISTS defect. Of the Yorkshire boars, 0.8% were afflicted with the KA defect and 7.6% with the ISTS defect. Boars diagnosed with the ISTS defect produced no litters. Fertility data were available from two artificially inseminated (AI) boars and six farm breeding boars affected with the KA defect. Breeding boars with 45-81% knobbed spermatozoa (n = 6) did not produce any litters out of 71 sows bred. AI boars with 25-30% knobbed spermatozoa had a poor non-return rate (on average 47% compared with 85% for normal control boars) and produced small litters, on average 2.5 piglets less than other boars of the same breed. Morphometry of testicular tissue and distribution of different cells in the seminiferous tubules were examined in nine boars. Boars with the KA defect had a smaller diameter of the seminiferous tubules (p < 0.05) and a lower number of Sertoli cells (p < 0.05) than controls. ISTS boars, in turn, had a significantly lower number of elongated spermatids (p < 0.05), and they also produced on average only 12% of the spermatozoa of normal boars. The ISTS defect is a manifestation of an autosomal recessive disease caused by an insertion in the KPL2 gene in porcine chromosome 16. Although we tried to map the KA defect, its aetiology remains unclear. PMID- 18397190 TI - Expression of genes in the canine pre-implantation uterus and embryo: implications for an active role of the embryo before and during invasion. AB - The aim of the present study was to assess genes expressed in maternal uterine tissue and pre-implantation embryos which are presumably involved in maternal recognition and establishment of canine pregnancy. For this purpose, 10 pregnant bitches were ovariohysterectomized between days 10 and 12 after mating. Four non pregnant bitches served as controls. Early pregnancy was verified by flushing the uterine horns with PBS solution. The collected embryos (n = 60) were stored deep frozen (-80 degrees C). Uterine tissue was excised, snaps frozen in liquid nitrogen and homogenized using TRI Reagent. All embryos from one litter were thawed together and also homogenized in TRI Reagent. RT-PCR was performed to prove mRNA expression of progesterone receptor, key enzymes of the prostaglandin synthesis pathway, selected growth factors, cytokines, immune cell receptors, major histocompatibility complex (MHC) and matrix-metalloproteinases (MMP). Only pregnant uteri revealed the presence of mRNA for interferon (IFN)-gamma, IL-4 and CD-8, which resembles the milieu in humans and other mammalians. Similarly, in day 10 embryos, mRNA for transforming growth factor-beta, insulin-like growth factor-1,-2, hepatocyte growth factor, leukaemia inhibitor factor, tumour necrosis factor-alpha, interleukin-1beta,-6,-8, cyclooxygenase-2, CD4(+) cells, and MMP-2 and -9 were detected, but not MHC-I or -II. We therefore suppose that the canine embryo, like its human counterpart, actively initiates measures to prevent attacks from the maternal immune system to prepare its own adhesion, nidation, growth and further development. PMID- 18397191 TI - Analysing motility parameters on fresh bull semen could help to predict resistance to freezing: a preliminary study. AB - Batches of straws often need to be thrown away after freezing because of a too few number of motile or progressive sperm cells after thawing. Our objective was to evaluate the possibility to predict before freezing the quality of the semen after freezing/thawing. Computer-Aided Sperm Analysis was performed on motility parameters both before and after freezing of 20 ejaculates from different bulls. Significant variation between bulls was observed both before and after freezing for all the analysed traits (anova2; p < 0.001): proportion of motile (%mot) and progressive (%prog) sperm, velocity on the curved line (VCL), velocity on the straight line (VSL), velocity on the average path (VAP), linearity (VSL/VCL), beat cross frequency and average orientation change of the head. Freezing significantly altered the motility parameters and correlations were found between samples analysed before and after freezing (Pearson coefficient: R = 0.43-0.72; p < 0.05). The mean VAP, VSL and the %prog obtained before freezing were highly correlated to the %mot and %prog observed after freezing (R = 0.75-0.82; p < 0.001). Applying thresholds on mean VAP and VSL values allowed us to predict respectively 6 and 7 of nine batches that would be rejected after freezing due to a too low %prog (<15%). Combining different traits did not add to the precision. In conclusion, analysis of velocity traits on fresh sperm seems more efficient than analysis of %mot or %prog to discard batches that will be of poor quality after freezing. PMID- 18397192 TI - Serum leptin concentrations in Lipizzan fillies. AB - To determine seasonal changes in circulating leptin concentrations, as well as leptin fluctuations in various patterns of seasonal ovarian activity, blood samples were taken in 107 Lipizzan fillies aged 1-4 years over the whole year from January to December. Serum concentrations were measured in samples obtained once a month (leptin) or twice a month (progesterone), and evaluated with respect to the age of animals (yearlings, 2-, 3- and 4-year olds) and the pattern of ovarian activity (anoestrous, seasonal cycle, prolonged cyclicity, continuous cyclicity). The mean and monthly leptin concentrations in 3- and 4-year olds were significantly higher than in yearlings and 2-year olds (p < 0.001). Circulating leptin levels in 3- and 4-year olds increased significantly in summer, to peak in August and then decline significantly (p < 0.001) in autumn. The fluctuations in younger fillies were less evident. The highest mean serum leptin concentrations were observed in fillies exhibiting continuous cyclicity and the lowest in those, exhibiting seasonal cyclicity (p < 0.001). In anoestrous fillies, leptin concentrations were lower than in fillies with prolonged or continuous cyclicity but higher than in fillies with seasonal cyclic activity (p > 0.05). PMID- 18397193 TI - Effects of functional exercise training on performance and muscle strength after meniscectomy: a randomized trial. AB - Muscular deficits and functional limitations have been found years after meniscectomy of the knee. The purpose of this randomized controlled trial was to examine the effect of functional exercise training on functional performance and isokinetic thigh muscle strength in middle-aged patients subsequent to meniscectomy for a degenerative tear. Four years after meniscectomy, 45 patients (29 men, 16 women) were randomized to functional exercise training, supervised by a physical therapist, three times weekly for 4 months or to no intervention. The exercise program comprised of postural stability training and functional strength and endurance exercises for leg and trunk muscles. Outcomes were three functional performance tests and isokinetic muscle strength. Thirty patients (16 exercisers/14 controls) completed the study. Compared with control patients, the exercise group showed significant improvement in one-leg hop (change 8 vs 2 cm; P=0.040), hamstrings strength 60 degrees /s (P=0.033), and quadriceps endurance 180 degrees /s (P=0.001). Functional exercise training was well tolerated and improved functional performance and thigh muscle strength in this group of middle aged subjects with a previous degenerative meniscal injury and partial meniscectomy. PMID- 18397194 TI - Cell biological and biomechanical evaluation of two different fixation techniques for rotator cuff repair. AB - Our objective was to evaluate the cell biology and biomechanical aspects of the healing process after two different techniques in open rotator cuff surgery - double-loaded bio-absorbable suture anchors combined with so-called arthroscopic Mason-Allen stitches (AAMA) and a trans-osseous suture technique combined with traditional modified Mason-Allen stitches (SMMA). Thirty-six mature sheep were randomized into two repair groups. After 6, 12, or 26 weeks, evaluation of the reinsertion site of the infraspinatus tendon was performed. The mechanical load to-failure and stiffness results did not indicate a significant difference between the two groups. After 26 weeks, fibrocartilage was sparse in the AAMA group, whereas the SMMA group showed the most pronounced amount of fibrocartilage. We found no ultrastructural differences in collagen fiber organization between the two groups. The relative expression of collagen type II mRNA in the normal group was 1.11. For the AAMA group, 6 weeks after surgery, the relative expression was 55.47, whereas for the SMMA group it was 1.90. This in vivo study showed that the AAMA group exhibited a tendon-to-bone healing process more favorable in its cell biology than that of the traditional SMMA technique. Therefore, the AAMA technique might also be more appropriate for arthroscopic repair. PMID- 18397195 TI - Treatment of longstanding groin pain in athletes: a systematic review. AB - The aims of this study were to determine (1) the kinds of treatments applied for longstanding groin pain (LGP) in athletes; (2) the results; and (3) the levels of evidence for the interventions. Digital databases P were searched for articles describing the effects of interventions for LGP in athletes. Treatment of LGP in athletes can consist of conservative measures such as rest or restricted activity, active or passive physical therapy, steroid injections or dextrose prolotherapy. Studies describing surgery generally mention failure of conservative measures, although a description of these conservative measures is mostly lacking. During surgery, a reinforcement of the abdominal wall is applied in most cases, using an open or laparoscopic approach. There is level I evidence that physical therapy aiming at strengthening and coordinating the muscles stabilizing hip and pelvis has superior results compared with passive physical therapy. For patients with a positive herniography and/or positive ilioinguinal or iliohypogastric nerve block tests, there are indications (level II) that surgery results in earlier return to sport compared with exercise therapy. Possibly, laparoscopic intervention might result in an earlier return to sport compared with open approach surgery (level III). For different clinical diagnoses, the same or similar surgical interventions were performed. PMID- 18397196 TI - Physiological demands of competitive basketball. AB - The aim of this study was to assess physiological demands of competitive basketball by measuring oxygen consumption (VO2) and other variables during practice games. Each of 12 players (20.4 +/- 1.1 years) was monitored in a 20-min practice game, which was conducted in the same way as actual games with the presence of referees and coaches. VO2 was measured by a portable system during the game and blood lactate concentration (LA) was measured in brief breaks. Subjects were also videotaped for time-motion analysis. Female and male players demonstrated respective VO2 of 33.4 +/- 4.0 and 36.9 +/- 2.6 mL/kg/min and LA of 3.2 +/- 0.9 and 4.2 +/- 1.3 mmol/L in the practice games (P>0.05). They spent 34.1% of play time running and jumping, 56.8% walking, and 9.0% standing. Pre obtained VO(2max) was correlated to VO(2) during play (r=0.673) and to percent of duration for running and jumping (r=0.935 and 0.962 for females and males, respectively). This study demonstrated a greater oxygen uptake for competitive basketball than that estimated based on a previous compendium. The correlation between aerobic capacity and activity level suggests the potential benefit of aerobic conditioning in basketball. PMID- 18397197 TI - TLR7/8 ligand, R-848, inhibits IgE synthesis by acting directly on B lymphocytes. AB - TLRs are involved in the regulation of immune responses. R-848, a TLR7/8 ligand, has potent anti-viral and anti-tumour properties and has been used as a new immune response modifier for enhancing Th1 immune response. In this study, we found that R-848 significantly inhibited IgE synthesis from murine B cells at the single cell levels by anti-CD40 plus IL-4-stimulated splenocytes, in which R-848 acted on the early stage of B cell differentiation to modulate IgE synthesis. This inhibitory effect of R-848 on IgE synthesis was not isotype specific as it also inhibited IgG1 synthesis. Moreover, R-848 had no significant effect on the production of IgG2a by anti-CD40 plus IL-4-stimulated splenocytes. Further studies showed that R-848 markedly promoted murine B cell activation induced by anti-CD40 plus IL-4 by up-regulating the expression of B cell activation markers CD25, CD69 and co-stimulatory molecule CD80. In contrast, R-848 inhibited the proliferation and division of murine B cells in anti-CD40 plus IL-4-stimulated splenocytes. R-848 promoted the production of IFN-gamma and IL-12 that were partially responsible for its inhibitory effect on IgE production by anti-CD40 plus IL-4 because the addition of anti-IFN-gamma or anti-IL-12 mAbs to the cultures could significantly restore IgE production by splenocytes. Importantly, R-848 had a direct effect on purified B cells to inhibit IgE production induced by anti-CD40 plus IL-4. Taken together, these results demonstrate that R-848 markedly inhibits IgE synthesis, and suggest that R-848 could be used to treat allergic diseases. PMID- 18397198 TI - Gender variation in interleukin-13 production: a possible mechanism of differential in vivo growth of a T-cell lymphoma. AB - Interleukin (IL)-13 is a T(H)2 type of cytokine that plays a crucial role in the pathophysiology of different type of infections, autoimmune diseases and malignancies. It has been shown to dampen the T(H)1 type of immune responses and favours tumour growth. In the present investigation, we have determined IL-13 level in serum and ascitic fluid in both the sexes of BALB/c strain of mice bearing a T-cell lymphoma of spontaneous origin, designated as Dalton's Lymphoma (DL). Further, we have studied the involvement of gender hormones on the IL-13 level and NKT-cell production of IL-13. It has been observed that there exists a gender variation or gender dimorphism in the IL-13 production. IL-13 level in serum is directly correlated with in vivo progressive growth of DL cells. We observed a tendency in female DL-bearing mice to have higher serum IL-13 level and faster growth of DL cells. This study, therefore, indicates that sex hormones are directly involved in the differential production of IL-13 that may be a factor responsible, at least in part, for the differential in vivo progressive growth of a T-cell lymphoma. PMID- 18397199 TI - Developing antibacterial vaccines in genomics and proteomics era. AB - Infectious diseases are the greatest cause of global morbidity and mortality. About half of this burden is caused by pathogenic bacteria. One of the most effective ways to prevent infectious diseases is to use vaccines. Limitations in the traditional approaches may be one of the reasons why vaccines are yet not available against some infectious bacterial agents. Recent advancements in high throughput 'omics' technologies and the availability of complete genome sequences of microbial pathogens and multiple isolates of the same species have dramatically changed the time frame and scope for identifying novel vaccine candidates. At present, 582 microbial genomes have been sequenced and sequencing of 1733 is in progress. There are more than 50 bacterial species for which the genome sequence is available for three or more isolates. Development of tools for in silico analysis has led to the identification of novel virulence genes, metabolic pathways and cell surface proteins that represent potential new targets for anti-microbial drug and vaccine strategies. In this review, we provide an overview of the 'omics'-based techniques that can be used to advance and accelerate the discovery of vaccine candidates against extracellular bacterial pathogens. By citing specific examples, we discuss how high-throughput molecular profiling techniques, such as genomics, transcriptomics and proteomics have contributed to the discovery of novel vaccine candidates. We end by contemplating on the emerging technologies that are likely to have a high impact on the field of vaccinology in the near future. PMID- 18397200 TI - The tuberculin skin test increases the responses measured by T cell interferon gamma release assays. AB - RUTI is a vaccine consisting of Mycobacterium tuberculosis bacilli grown in stress conditions that is fragmented, detoxified and liposomed. RUTI was designed to shorten the treatment of latent tuberculosis infection (LTBI) with isoniazid from 9 months to just 1 month, by additional treatment with two inoculations of RUTI 4 weeks apart. During the validation process for monitoring the immunogenicity of administration of RUTI in a Phase I clinical trial, the question arose whether to introduce the tuberculin skin test (TST) in the screening of non-LTBI volunteers. This study was designed to evaluate the effect of TST on subsequent different T-cell interferon-gamma release assay (TIGRA) responses, using a spectrum of M. tuberculosis-related antigens (ESAT-6, CFP-10, 16 kDa, 19 kDa, MPT64, Ag 85B, 38 kDa, hsp65, PPD and BCG). The results showed an increase in post-TST response even in non-LTBI subjects for most antigens tested, as measured both by whole blood assay (WBA) and ELISPOT. Increased ELISPOT response decreased toward pre-TST levels within 1 month whereas the WBA response did not. Taking into account that there is no definitive correlation between TST and TIGRA tests to diagnose LTBI and the feasibility that TST might alter the immune monitoring included in clinical trials, these data suggest that TST determination should be carefully planned to avoid any interference with TIGRA. PMID- 18397201 TI - Antifungal immunotherapy and immunomodulation: a double-hitter approach to deal with invasive fungal infections. AB - In recent years, the incidence of life-threatening fungal infections has dramatically increased. Despite significant developments in antifungal chemotherapy, its efficacy remains limited by the inability to sterilize infected organs and the tendency to induce resistance and cause side effects. In response to these challenges, it is now recognized that several aspects of antifungal immunity can be modulated to better deal with fungal infections. Extensive work was carried out on the development and testing of preventive and therapeutic antifungal vaccines. The potential use of cytokines, adoptive T-cell transfer, monoclonal antibodies (MoAb) and antimicrobial peptides (AMP) as solo or adjunctive therapies is also receiving much attention. Although each of these immune-based treatment strategies has many advantages and some shortcomings, none on its own, has proven satisfactorily effective to deal with invasive fungal infections. Appropriate combinations that optimize the advantages and minimize the disadvantages of immune-based antifungals are still lacking mainly due to the immense difficulty in sorting out candidate combinations given the long list of choices. In this review, immune-based antifungals are divided into two general categories on the basis of the intended target being the host (immunomodulation through vaccines, cytokines, adoptive T-cell transfer or MoAb) or the pathogen (immunotherapy through MoAb or AMP). Potential advantages and disadvantages of immunotherapy and immunomodulation are tentatively discussed so as to facilitate the design of future studies that aim at devising more potent immune-based antifungal treatment combinations. PMID- 18397202 TI - Hepatitis B virus infection and the dialysis patient. AB - Prevention of nosocomial transmission of hepatitis B virus (HBV) has been a signal achievement in the management of chronic kidney disease. The rate of serum hepatitis B surface antigen (HBsAg) seropositivity in patients on maintenance hemodialysis in the developed world is currently low (0-10%) but outbreaks of acute HBV infection continue to occur in this setting. The prevalence of HBV infection within dialysis units in developing countries appears higher (2-20%) based on relatively few reports. Although data are limited, HBV infection in dialysis population diminishes survival; HBV viral load in HBsAg-positive dialysis patients is reportedly low and stable over time. Updated recommendations for the management of HBsAg chronic carriers on maintenance dialysis have been issued. No rigorously controlled treatment trials for treatment of hepatitis B with either interferon or lamivudine therapy in dialysis patients are currently available. PMID- 18397203 TI - Time to symptomatic vascular stenosis at different locations in patients with arteriovenous grafts. AB - Most arteriovenous grafts fail due to irreversible thrombosis, superimposed on hemodynamically significant vascular stenosis. Previous studies observed the highest frequency of stenosis at the venous anastomosis, without addressing the timing of stenosis. The present study quantified time to symptomatic stenosis at different vascular locations, and related it to permanent graft failure. A prospective computerized vascular access database was queried retrospectively to identify 309 hemodialysis patients receiving new upper extremity grafts during a 4-year period at a large dialysis center. For each vascular site we calculated the time to symptomatic stenosis using survival techniques. The cumulative likelihood of symptomatic stenosis at 2 years was 67% for venous anastomotic stenosis, 19% for intra-graft stenosis, 16% for venous outlet stenosis, 13% for central vein stenosis, and 5% for arterial anastomotic stenosis. The cumulative risk of graft failure at 2 years was 40%. Stenosis at the venous anastomosis was twice as likely as cumulative graft failure (hazard ratio [HR] 1.95; 95% confidence interval [CI], 1.65-2.52, p < 0.001). In contrast, intra-graft stenosis was half as likely as cumulative graft failure (HR 0.45; 95% CI, 0.36 0.61, p < 0.001). Central vein stenosis was more likely in patients with a previous ipsilateral catheter compared with those without one (HR 2.40; 95% CI, 1.39-5.58, p = 0.004). Symptomatic stenosis occurs much earlier at the venous anastomosis compared with other vascular sites. Moreover, preexisting ipsilateral internal jugular dialysis catheters more than double the risk of central vein stenosis. PMID- 18397204 TI - Thrombosis with erythropoietic stimulating agents-does iron-deficient erythropoiesis play a role? AB - Thrombocytosis is common in iron deficiency and resolves following iron repletion. Increased platelet number, whether from iron deficiency or from other causes, may increase the risk of thrombovascular events. One mechanism thought to mediate iron deficiency-induced thrombocytosis is increased erythropoietin production. Similarly, erythropoietic stimulating agents (ESA) have long been known to increase platelet number and frequently lead to functional or absolute iron deficiency. This state of relative or absolute iron deficiency may be the mechanism whereby ESA increase the platelet count. If correct, co-administration of iron should prevent or diminish ESA-driven thrombocytosis. Data from the DRIVE trial in hemodialysis patients do, in fact, suggest that this is the case. Platelet counts in patients receiving IV iron decreased, while they remained unchanged in patients not given iron (mean change -29,000/microl vs. -0/microl; p = 0.017). Other supporting data have been observed in IV iron trials in oncology patients. The harm from higher hemoglobin targets and higher ESA doses may be mediated in part through induction of iron deficiency and thrombocytosis. The major anemia trials of ESAs have not reported platelet data, but should examine the relationship of platelet count, iron deficiency, IV iron administration, and cardiovascular events in greater detail. PMID- 18397205 TI - Obesity as a predictor of vascular access outcomes: analysis of the USRDS DMMS Wave II study. AB - Arteriovenous fistulae (AVF) are widely regarded as the preferred vascular access in hemodialysis patients due to their primary patency and patient survival benefits. While the obesity paradox has been associated with improved cardiovascular morbidity and all-cause mortality in dialysis patients, its long term vascular access outcomes are less clear. Recent literature has suggested that obese patients may have increased early and late fistula failure. The purpose of this study was to explore the relationships between obesity and vascular access outcomes. We performed a retrospective cohort analysis using the USRDS DMMS Wave 2 data set. All incident dialysis patients as of January 1, 1996, over the age of 18, receiving only hemodialysis as mode of renal replacement therapy were eligible for inclusion. Among other variables, data collected for the DMMS Wave 2 included: type and location of vascular access, AVF maturity, vascular access revision, and failure. Logistic regression analyses were used to examine the relationships between obesity and vascular access outcomes, adjusting for important covariates. In all, 1486 hemodialysis patients were included. Using body mass index (BMI) <30 kg/m(2) as reference, obesity did not emerge as a factor in predicting vascular access revisions or failures. An increased risk of AVF failure to mature was found only in the highest BMI quartile (>or=35 kg/m(2)) (aOR 3.66 [95% CI 1.27-10.55], p = 0.017). Peripheral vascular disease was independently associated with an increased risk of AVF failure (aOR 2.78 [95% CI 1.01-7.63], p = 0.047) and arteriovenous graft (AVG) failure (aOR 1.65 [95% CI 1.03-2.64], p = 0.036). Obesity was not associated with increased AVF or AVG revision rates or failure and only associated with poorer AVF maturity at highest BMI quartile. We conclude that obesity should not preclude placement of AVF as vascular access of choice, except in the very obese where assessment should be individually based. PMID- 18397206 TI - Constitutive expression of Arabidopsis NPR1 confers enhanced resistance to the early instars of Spodoptera litura in transgenic tobacco. AB - In Arabidopsis, NPR1 (AtNPR1) regulates salicylic acid (SA)-mediated activation of PR genes at the onset of systemic acquired resistance. AtNPR1 also modulates SA-induced suppression of jasmonic acid-responsive gene expression, and npr1 mutants manifest enhanced herbivore resistance. We have raised stable transgenic tobacco lines, expressing AtNPR1 constitutively, which showed elevated expression of PR1 and PR2 genes upon SA treatment. Herbivore bioassays with a generalist polyphagous pest, Spodoptera litura, revealed that the transgenic lines exhibited enhanced resistance compared to the wild-type plants, particularly with respect to younger larval populations. Insect-mediated injury induced several protease inhibitors (PIs), more significantly a 40-kDa serine PI in all the tobacco lines, but the induction was higher in the transgenic plants. We show in this communication that heterologous expression of AtNPR1 provides enhanced resistance to early larval populations of the herbivore, Spodoptera in transgenic tobacco plants. PMID- 18397207 TI - Analysis of nodulation kinetics in Frankia-Discaria trinervis symbiosis reveals different factors involved in the nodulation process. AB - The induction of root nodule development in actinorhizal symbiosis would depend on the concentration of factors produced by the bacteria and the plant. A detailed analysis of nodulation description parameters revealed different factors related to the nodulation process. The initial time for nodulation (t(0)), the initial nodulation rate (v(0)) and the total time of nodule development (t(NOD)) were defined and consequently quantified in different experimental conditions: co inoculation of Discaria trinervis with increasing concentrations of different non infective bacteria together with the full compatible infective Frankia strain (the indicator strain) used at a limiting concentration or by changing plant factor(s) concentration. All the above nodulation parameters were modified by changing doses of full compatibility infective strain Frankia BCU110501; v(0) appears to be an expression of symbiotic recognition between partners as only fully symbiotic indicator Frankia BCU110501 was able to change it; t(0) seems not to reflect symbiotic recognition because it can also be modified by non-infective Frankia but suggest the existence of a basic level of plant microbe recognition. The initial time for nodulation t(0), reflecting the time required for the early interactions toward nodulation, is an inverse measure of the ability to establish early interactions toward nodulation. The increase in plant factors concentration also reduces t(0) values, suggesting that a plant factor is involved and favors very early interactions. Increases in plant factors concentration also modify the final number of nodules per plant and the nodule cluster profile along the taproot as an expression of the autoregulation phenomenon. Meanwhile, Frankia inoculums' concentration, either infective or not, modified t(NOD) in an opposite way plant factors did. In conclusion, the analysis of nodulation kinetics appears to be an appropriate tool to investigate factors involved in the symbiotic interaction leading to the formation of nitrogen-fixing nodules. PMID- 18397208 TI - Crops and genotypes differ in efficiency of potassium uptake and use. AB - Cultivars with increased efficiency of uptake and utilization of soil nutrients are likely to have positive environmental effects through reduced usage of chemicals in agriculture. This review assesses the available literature on differential uptake and utilization efficiency of K in farming systems. Large areas of agricultural land in the world are deficient in K (e.g. 3/4 of paddy soils in China, 2/3 of the wheatbelt in Southern Australia), with export in agricultural produce (especially hay) and leaching (especially in sandy soils) contributing to lowering of K content in the soil. The capacity of a genotype to grow and yield well in soils low in available K is K efficiency. Genotypic differences in efficiency of K uptake and utilization have been reported for all major economically important plants. The K-efficient phenotype is a complex one comprising a mixture of uptake and utilization efficiency mechanisms. Differential exudation of organic compounds to facilitate release of non exchangeable K is one of the mechanisms of differential K uptake efficiency. Genotypes efficient in K uptake may have a larger surface area of contact between roots and soil and increased uptake at the root-soil interface to maintain a larger diffusive gradient towards roots. Better translocation of K into different organs, greater capacity to maintain cytosolic K(+) concentration within optimal ranges and increased capacity to substitute Na(+) for K(+) are the main mechanisms underlying K utilization efficiency. Further breeding for increased K efficiency will be dependent on identification of suitable markers and compounding of efficiency mechanisms into locally adapted germplasm. PMID- 18397209 TI - SLC24A5: exchanging genetic and biochemical knowledge. PMID- 18397210 TI - Perspectives on the structure and function of melanin. PMID- 18397211 TI - Immune response to rabies vaccination in pediatric transplant patients. AB - Children have become engaged in a wider variety of activities as the success of solid organ transplantation has improved. These activities can result in exposure to infectious agents for which there are no data documenting the efficacy of standard treatment in children on immunosuppressive therapy. This is a retrospective review of five OLT patients and three RT patients who were potentially exposed to rabies during camp. They completed the immunoprophylaxis treatment for rabies exposure outlined by the CDC in the 2003 Red Book. Rabies titers were followed for six to 12 months post-immunization. All five OLT patients were on tacrolimus. All three RT patients were on tacrolimus, mycophenolate mofetil, and prednisone. At the time of exposure median age was 10.0 yr (8.4-17.3). None of the subjects developed rabies. A positive rabies titer, indicative of successful immunization, was present by one month in seven subjects and all subjects by six months. Rabies vaccination in pediatric transplant patients is safe and associated with the successful production of antirabies titers. PMID- 18397212 TI - A conditioning regimen of busulfan, fludarabine, and melphalan for allogeneic stem cell transplantation in children with juvenile myelomonocytic leukemia. AB - A pilot study was undertaken using a myeloablative conditioning with fludarabine, busulfan, and melphalan to improve the outcome of HSCT in 10 children, aged six months to six yr, with JMML. All patients were conditioned with oral busulfan (560 mg/m(2)), fludarabine (120 mg/m(2)), and melphalan (180-210 mg/m(2)) prior to HSCT, and received stem cells from bone marrow in seven cases, and from cord blood in three cases. Engraftment was documented in eight patients, whereas graft failure occurred in two, one of whom had received HLA-mismatched cord blood and other had received bone marrow from HLA-mismatched mother. Three patients, including two in who graft failure had occurred, relapsed. Five patients developed acute GVHD and two developed chronic GVHD. Seven patients are alive and in remission 27-69 months after transplantation. Thus, our study showed that HSCT following conditioning with fludarabine, busulfan, and melphalan was well tolerated and appeared to be effective for JMML. PMID- 18397213 TI - Aspergillus fumigatus endocarditis in a pediatric liver transplant recipient: favorable outcome without cardiac surgery. AB - Transplant recipients are very susceptible to invasive aspergillosis, which increases mortality rate. Disseminated aspergillosis in the liver transplant recipient can affect virtually any organ and endocarditis is often lethal despite cardiac surgery and antifungal therapy. We report the case of a eight-month-old girl who presented with Aspergillus fumigatus endocarditis 18 days after liver transplantation that was successfully treated by a combination of antifungal drugs associated to a low dosage of immunosuppressive therapy. PMID- 18397214 TI - Recurrent pancreatitis after renal transplantation in a child. AB - A five-yr-old girl, who was a renal transplant recipient, presented with nausea, vomiting, epigastric discomfort, papules, and vesicles on her body. She was diagnosed with acute pancreatitis and varicella zoster infection because her serum amylase and lipase levels were positive. Fourteen months later, she was readmitted with nausea, vomiting, and epigastric pain similar to the previous symptoms and was diagnosed with acute pancreatitis. This case report indicates that acute pancreatitis can be one of a number of complications following pediatric renal transplantation and can recur because of various causes. PMID- 18397215 TI - Percutaneous retrograde colangiography of segment III bile duct, through the previously catheterized segment II bile duct, in 15-month-old patient after partial liver transplantation with two separate bilio-enteric anastomoses. AB - We report one case of percutaneous retrograde colangiography of segment III bile duct, through the previously catheterized segment II bile duct, in 15-month-old patient after partial liver transplantation and two separate biliary anastomoses. The retrograde diagnostic cholangiogram confirmed the suspicion of anastomotic stricture and distended the bile ducts, which were easily punctured with percutaneous approach with a 20 Ga Chiba needle. PMID- 18397216 TI - Chronic high Epstein-Barr viral load carriage in pediatric liver transplant recipients. AB - Development of EBV disease and PTLD is usually accompanied by the detection of a high EBV load in peripheral blood. However, many children undergoing primary EBV infection following LTx will maintain chronically elevated EBV loads in the absence of clinical symptoms. To better understand this phenomenon, we retrospectively reviewed the records of children undergoing LTx at our center from 1997 to 2007 to identify chronic high EBV load carriers in this population. A CHL state was defined by the presence of a high load for >50% of samples for greater than or equal to six months following either asymptomatic or complete clinical resolution of EBV disease/PTLD. A total of 35 CHL carriers were identified. Pretransplant serologies were available for 29 of the 35; 22/29 (76%) were EBV negative prior to LTx; eight of these 22 developed their CHL state at the time of their primary EBV infection. Fourteen of the 35 had EBV disease (n = 7) or PTLD (n = 7) prior to development of the CHL state. Only one of 35 CHL carriers developed PTLD or lymphoma while they were a high load carrier. In all, 23/35 resolved their CHL state without apparent sequelae while 11 children continue to be asymptomatic high load carriers. These data provide important information about the outcome of chronic EBV high load carriage in pediatric liver transplant recipients. PMID- 18397217 TI - Mountain coniferous forests, refugia and butterflies. AB - The boreal coniferous forests form the most extended vegetation zone of the Northern Hemisphere. As opposed to North America, they are disconnected from the mountain coniferous forests in Europe, because of the dominant east-west direction of the mountain chains. Consequently, the mountain forests show some unique characteristic features of glacial survival and postglacial history, as well. The mountain coniferous forests have numerous common floral and faunal elements with the boreal zone. However, the few unique faunal elements of the European mountain coniferous forests can be used to unravel the peculiar patterns and processes of this biome. In this issue of Molecular Ecology, Thomas Schmitt and Karola Haubrich (2008) use the relatively common and taxonomically well studied butterfly, the large ringlet (Erebia euryale) to identify the last glacial refugia and postglacial expansion routes. PMID- 18397218 TI - Habitat fragmentation and genetic diversity of an endangered, migratory songbird, the golden-cheeked warbler (Dendroica chrysoparia). AB - Landscape genetic approaches offer the promise of increasing our understanding of the influence of habitat features on genetic structure. We assessed the genetic diversity of the endangered golden-cheeked warbler (Dendroica chrysoparia) across their breeding range in central Texas and evaluated the role of habitat loss and fragmentation in shaping the population structure of the species. We determined genotypes across nine microsatellite loci of 109 individuals from seven sites representing the major breeding concentrations of the species. No evidence of a recent population bottleneck was found. Differences in allele frequencies were highly significant among sites. The sampled sites do not appear to represent isolated lineages requiring protection as separate management units, although the amount of current gene flow is insufficient to prevent genetic differentiation. Measures of genetic differentiation were negatively associated with habitat connectivity and the percentage of forest cover between sites, and positively associated with geographic distance and the percentage of agricultural land between sites. The northernmost site was the most genetically differentiated and was isolated from other sites by agricultural lands. Fragmentation of breeding habitat may represent barriers to dispersal of birds which would pose no barrier to movement during other activities such as migration. PMID- 18397219 TI - Mitochondrial DNA under siege in avian phylogeography. AB - Mitochondrial DNA (mtDNA) has been the workhorse of research in phylogeography for almost two decades. However, concerns with basing evolutionary interpretations on mtDNA results alone have been voiced since the inception of such studies. Recently, some authors have suggested that the potential problems with mtDNA are so great that inferences about population structure and species limits are unwarranted unless corroborated by other evidence, usually in the form of nuclear gene data. Here we review the relative merits of mitochondrial and nuclear phylogeographical studies, using birds as an exemplar class of organisms. A review of population demographic and genetic theory indicates that mitochondrial and nuclear phylogeographical results ought to concur for both geographically unstructured populations and for populations that have long histories of isolation. However, a relatively common occurrence will be shallow, but geographically structured mtDNA trees--without nuclear gene corroboration- for populations with relatively shorter periods of isolation. This is expected because of the longer coalescence times of nuclear genes (approximately four times that of mtDNA); such cases do not contradict the mtDNA inference of recent isolation and evolutionary divergence. Rather, the nuclear markers are more lagging indicators of changes in population structure. A review of the recent literature on birds reveals the existence of relatively few cases in which nuclear markers contradict mitochondrial markers in a fashion not consistent with coalescent theory. Preliminary information from nuclear genes suggests that mtDNA patterns will prove to be robust indicators of patterns of population history and species limits. At equilibrium, mitochondrial loci are generally a more sensitive indicator of population structure than are nuclear loci, and mitochondrial estimates of F(ST)-like statistics are generally expected to exceed nuclear ones. Hence, invoking behavioural or ecological explanations of such differences is not parsimonious. Nuclear genes will prove important for quantitative estimates of the depths of haplotype trees, rates of population growth and values of gene flow. PMID- 18397220 TI - Modeling the inactivation kinetics of bacillus spores by glutaraldehyde. AB - AIMS: Our goal was to develop a mathematical kinetic model to predict the sporicidal activity of glutaraldehyde, which is an active ingredient frequently used in commercial products employed for liquid disinfection and decontamination. METHODS AND RESULTS: We used our previously published data on spore inactivation by glutaraldehyde to develop a predictive model obtained by calculating multiple independent modifying functions. The model was then validated by comparing model predicted values to new experimental data. For model validation, quality controlled spores of Bacillus athrophaeus (previously and generally known as Bacillus subtilis globigii) were exposed under conditions where several physicochemical variables were modified simultaneously, and the spore surviving fractions were measured by titration. CONCLUSIONS: The model predicted within one order of magnitude variations in sporicidal effectiveness due to changes in main parameters (glutaraldehyde concentration, temperature or time-duration of the treatment). Other parameters such pH, salinity and the effect of serum concentration were also addressed, albeit with less accuracy. SIGNIFICANCE AND IMPACT OF THE STUDY: The model should be useful to quantitatively estimate the effectiveness of glutaraldehyde-based disinfectants, decontaminants, and germicides under the described conditions, particularly when limited data are available or when spore virulence (like that of Bacillus anthracis) precludes extensive experimentation. A similar approach could predict the effectiveness of a variety of decontaminant and disinfecting agents. PMID- 18397222 TI - Hepatitis RNA viruses and hepatocellular carcinoma. PMID- 18397223 TI - Cause of death in individuals with chronic HBV and/or HCV infection, a nationwide community-based register study. AB - Studies on chronic viral hepatitis and mortality have often been made on selected populations or in high-endemic countries. The aim of this study was to investigate the causes of death and the mortality rates in the nationwide cohorts of people chronically infected with hepatitis B virus (HBV) and/or hepatitis C virus (HCV) in Sweden, a low-endemic country. All notifications on chronic HBV infection and HCV infection 1990-2003 were linked to the Cause of Death Register. A total of 9517 people with chronic HBV infection, 34 235 people with HCV infection and 1601 with chronic HBV-HCV co-infection were included, and the mean observation times were 6.4, 6.3 and 7.9 years, respectively. The mortality in the cohorts was compared with age- and gender-specific mortality in the general population and standardized mortality ratios (SMR) were calculated. All-cause mortality was significantly increased, SMR 2.3 (HBV), 5.8 (HCV) and 8.5 (HBV HCV), with a great excess liver-related mortality in all cohorts, SMR 21.7, 35.5 and 46.2, respectively. In HCV and HBV-HCV infected there was an increased mortality due to drug-related psychiatric diagnoses (SMR: 20.7 and 27.6) and external causes (SMR: 12.4 and 11.4), predominantly at younger age. To conclude, this study demonstrated an increased all-cause mortality, with a great excess mortality from liver disease, in all cohorts. In people with HCV infection the highest excess mortality in younger ages was from drug-related and external reasons. PMID- 18397224 TI - Hepatocellular carcinoma and other primary liver cancers in hepatitis C patients in Sweden - a low endemic country. AB - The aim of this study was to assess the risk of hepatocellular carcinoma (HCC) and other primary liver cancers (PLC) in the nationwide cohort of hepatitis C virus (HCV) infected patients in Sweden. The basis was the total HCV-cohort notified in 1990-2004, after excluding 3238 people also reported with hepatitis B, the study cohort consisted of 36 126 people contributing an observation time of 246 105 person-years. The most common route of transmission was intravenous drug use (57%). The national Cancer Registry was used for follow-up, and 354 developed PLC (mainly HCC), of whom 234 were eligible for statistical analysis. The PLC incidence in the HCV cohort was compared with the incidence in the general population, and a standardized incidence ratio (SIR) was calculated for six different strata according to estimated duration of infection. The highest relative risk, SIR: 46 (95% CI: 36-56) was found in the stratum 25-30 years with HCV infection and SIR: 40 (95% CI: 31-51) in the stratum 30-35 years with infection. In the entire community-based HCV cohort in Sweden we found a highly increased risk of liver cancer compared to the general population. The highest relative risk was among people who had been infected for more than 25 years. PMID- 18397225 TI - Effects of silymarin on the resolution of liver fibrosis induced by carbon tetrachloride in rats. AB - Silymarin, a standardized extract of the milk thistle (Silybum marianum), has a long tradition as a herbal remedy, and was introduced as a hepatoprotective agent a few years ago. However, the therapeutic effects of silymarin remain undefined. Carbon tetrachloride (CCl4) is a xenobiotic used extensively to induce oxidative stress and is one of the most widely used hepatic toxins for experimental induction of liver fibrosis in the laboratory. In this study, we investigated the restoration of the CCl4-induced hepatic fibrosis by high dose of silymarin in rats. After treatment with oil (as normal group; n = 6) or CCl4 [as model (n = 7) and therapeutic (n = 7) groups] by intragastric delivery for 8 weeks for the induction of liver fibrosis, the rats in the normal and model group were administered orally normal saline four times a week for 3 weeks whilst the therapeutic group received silymarin (200 mg/kg). The histopathological changes were observed with Masson staining. The results showed that the restoration of the CCl4-induced damage of liver fibrosis in the therapeutic group was significantly increased as compared to that in the model group. Moreover, silymarin significantly decreased the elevation of aspartate aminotransferase (AST), alanine aminotransferase, and alkaline phosphatase in serum, and also reversed the altered expressions of alpha-smooth muscle actin in liver tissue. Therefore, these findings indicated that silymarin may have the potential to increase the resolution of the CCl4-induced liver fibrosis in rats. PMID- 18397226 TI - Predictors of a sustained virological response in patients with genotype 4 chronic hepatitis C. AB - OBJECTIVES: To determine the clinical, biological, virological and histological predictive factors associated with a sustained virological response (SVR) to combined interferon therapy among Egyptian patients infected by genotype 4 hepatitis C virus (HCV). PATIENTS AND METHODS: Individual data from 250 patients with genotype 4 chronic hepatitis C, treated with different regimens of combined interferon, were analysed. The primary end point was SVR defined as undetectable HCV RNA by polymerase chain reaction (PCR) 24 weeks after the end of treatment. Multivariate logistic regression analysis was performed to select the independent prognostic parameters associated with SVR. RESULTS: A sustained virological response was achieved among 137/250 (54.8%) patients. Baseline factors independently and negatively associated with SVR were serum alpha-fetoprotein (AFP) level (above 0.3 upper limit of normal) [odds ratio (OR)=0.5, 95% confidence interval (CI): 0.2-0.8], severe fibrosis (Metavir score >F2) (OR=0.4, 95% CI: 0.2-0.8), presence of steatosis (OR=0.5, 95% CI: 0.3-0.97) and standard interferon treatment (OR=0.4, 95% CI: 0.2-0.8). CONCLUSIONS: Among genotype 4 chronic hepatitis C patients, severe fibrosis, severe steatosis, treatment with standard interferon and a high serum AFP level were all negatively associated with SVR. Pretreatment serum AFP level should be considered in the routine assessment of factors predictive of a treatment response. PMID- 18397227 TI - Resveratrol inhibits cell growth in a human cholangiocarcinoma cell line. AB - BACKGROUND/AIMS: Cholangiocarcinoma is a devastating tumour with a poor prognosis. An efficient therapy is unavailable in unoperable patients and new drugs are widely sought for and required. Resveratrol (RES) is a natural molecule with a reported anticancer effect, evaluated on different tumour cell lines. We tested the efficacy of RES on a cholangiocarcinoma cell line for the first time. METHODS: We used the human SK-ChA-1 cell line, cultured in the classical two dimensional model and in the three-dimensional spheroids. After RES exposure morphology, cell viability (colony-forming assay), lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and cancer antigen (CA) 19-9 medium releases, cellular transglutaminase activity, karyotype and cell cycle were evaluated. RESULTS: Resveratrol inhibited cell growth in both the cell culture systems used (from -15 to -80% vs untreated controls) and induced a 40-fold increase of LDH and ALP activities in the culture medium. Also, transglutaminase (TG) activity increased in the cell lysates, together with a cell cycle perturbation characterised by an accumulation in the G(1)/S phase. Karyotype and CA 19-9 expression were not influenced by the treatment. CONCLUSIONS: The observed cytotoxic effect of RES on the human cholangiocarcinoma SK-ChA-1 cell line cultured two- and three dimensionally suggests to further analyse its chemotherapic/chemopreventive possibilities for this kind of cancer. PMID- 18397228 TI - Upregulation of indoleamine 2,3-dioxygenase in hepatocyte during acute hepatitis caused by hepatitis B virus-specific cytotoxic T lymphocytes in vivo. AB - BACKGROUND/AIMS: Indoleamine-2,3-dioxygenase (IDO) is a tryptophan-catabolizing enzyme inducing suppression of T-cell function and immune tolerance. In hepatitis B virus (HBV) transgenic (Tg) mice, the adoptive transfer of HBV-specific cytotoxic T lymphocytes (CTL) causes a necroinflammatory liver disease that is histologically similar to acute viral hepatitis in man. The present study aimed to determine IDO expression in the liver and hepatocytes during an acute hepatitis model. METHODS: Serum l-kynurenine (l-Kyn) concentration in HBV Tg mice administered with HBV-specific CTL was measured over time, together with serum levels of alanine aminotransferase (ALT). Furthermore, we examined the expression of IDO in the total liver and isolated hepatocytes of HBV Tg mice after CTL injection using immunohistochemical analysis and reverse-transcription polymerase chain reaction (PCR). RESULTS: In HBV Tg mice, HBV-specific CTL induced, over the course of several days, a chronic increase in serum l-Kyn levels, which was associated with a sustained enhancement of liver IDO activity. In particular, IDO expression was enhanced in the liver parenchymal cells (hepatocytes) after HBV specific CTL injection both in immunohistochemical analysis and in reverse transcription PCR. Moreover, murine recombinant interferon-gamma (IFN-gamma) directly increased the IDO expression in primary hepatocytes in vitro. CONCLUSIONS: Cytotoxic T lymphocytes transduction results in the upregulation of IDO, which might downregulate T-cell responsiveness. Our findings provide evidence that hepatocyte itself expresses IDO and increases levels of l-Kyn in the blood in acute lethal hepatitis of mice. These data indicate that HBV infection facilitates the induction of IDO in response to proinflammatory cytokines, particularly IFN-gamma. PMID- 18397229 TI - Interferon-alpha treatment in children and young adults with chronic hepatitis B: a long-term follow-up study in Taiwan. AB - BACKGROUND/AIMS: The short- and long-term benefits of interferon (IFN)-alpha therapy in young patients with chronic hepatitis B (CHB) acquiring infection perinatally or during early childhood have been questioned. METHODS: Twenty-one Taiwanese hepatitis B envelope antigen (HBeAg)-positive CHB patients aged 1.8 21.8 years (median 14.0 years) with alanine aminotransferase (ALT)>80 IU/L at entry were enrolled for IFN-alpha therapy. They received IFN-alpha therapy with a dose of 3 MU/m(2)/day three times a week for 24 weeks. A control group included untreated 21 CHB patients closely matched for gender, age, duration of ALT >80 IU/L and HBeAg status. All 42 patients were prospectively followed for 6.5-12.5 years after the end of therapy. RESULTS: The cumulative rate of virological response [anti-HBe seroconversion and serum hepatitis B virus (HBV)-DNA <10(5) copies/ml] was not different between the IFN-treated patients and control patients at 1 year (41 vs 44%) and at 6 years (88 vs 89%) after stopping treatment. Serum hepatitis B surface antigen loss occurred in two (9.5%) treated patients and in one (4.8%) control patient. Patients with a successful treatment response (anti-HBe seroconversion, HBV-DNA <10(2) copies/ml and ALT normalization at 1 year after stopping treatment) were younger than those without a successful response (P=0.03). A lower pretreatment serum HBV-DNA level (<2 x 10(8) copies/ml) is not only a significant factor to predict successful treatment response (P=0.008) but also has a beneficial effect on the long-term cumulative rate of virological response in IFN-treated patients (P=0.021), but not in control patients. Genotype difference or emergence of a precore stop codon mutant before treatment was not predictive for HBeAg clearance. CONCLUSION: For young CHB patients in Taiwan with infection occurring perinatally or in early childhood, the real advantage of IFN-alpha therapy was not observed. IFN-alpha therapy showed a beneficial effect on short- and long-term virological outcomes only in those with a lower pretreatment serum HBV-DNA level. PMID- 18397230 TI - All-trans-retinoic acid ameliorates carbon tetrachloride-induced liver fibrosis in mice through modulating cytokine production. AB - BACKGROUND/AIMS: Liver fibrosis with any aetiology, induced by the transdifferentiation and proliferation of hepatic stellate cells (HSCs) to produce collagen, is characterized by progressive worsening in liver function, leading to a high incidence of death. We have recently reported that all-trans retinoic acid (ATRA) suppresses the transdifferentiation and proliferation of lung fibroblasts and prevents radiation- or bleomycin-induced lung fibrosis. METHODS: We examined the impact of ATRA on carbon tetrachloride (CCl(4))-induced liver fibrosis. We performed histological examinations and quantitative measurements of transforming growth factor (TGF)-beta1 and interleukin (IL)-6 in CCl(4)-treated mouse liver tissues with or without the administration of ATRA, and investigated the effect of ATRA on the production of the cytokines in quiescent and activated HSCs. RESULTS: CCl(4)-induced liver fibrosis was attenuated in histology by intraperitoneal administration of ATRA, and the overall survival rate at 12 weeks was 26.5% without ATRA (n=25), whereas it was 75.0% (n=24) in the treatment group (P=0.0187). In vitro studies disclosed that the administration of ATRA reduced (i) the production of TGF-beta1, IL-6 and collagen from HSCs, (ii) TGF-beta-dependent transdifferentiation of the cells and IL-6-dependent cell proliferation and (iii) the activities of nuclear factor kappaB p65 and p38mitogen-activated protein kinase, which stimulate the production of TGF-beta1 and IL-6, which could be the mechanism underlying the preventive effect of ATRA on liver fibrosis. CONCLUSIONS: Our findings indicate that ATRA ameliorates liver fibrosis. As the oral administration of the drug results in good compliance, ATRA could be a novel approach in the treatment of liver fibrosis. PMID- 18397231 TI - Insulin resistance and hepatitis C virus: a case-control study of non-obese, non alcoholic and non-steatotic hepatitis virus carriers with persistently normal serum aminotransferase. AB - BACKGROUND/AIMS: Recent studies using transgenic mouse models have demonstrated that the presence of hepatitis C virus (HCV) singularly induces insulin resistance (IR). When evaluated in humans, the exclusion of other factors influencing IR, such as obesity, alcohol intake, hepatic inflammation and steatosis is needed, but only few studies have been performed to these ends. Therefore, we aimed at exploring the singular effects of HCV on glucose metabolism through analysis of HCV carriers with persistently normal serum aminotransferase. METHODS: Non-obese, non-diabetic and non-alcoholic HCV carriers (n=30) were enrolled with 30 hepatitis B virus carriers matched by age, gender, body mass index and waist-to-hip ratio. All patients maintained normal serum aminotransferase (<30 U/L), hyaluronic acid (<50 ng/ml) and platelet count (>150 x 10(3)/microl) for more than 5 years without additional treatments, and had no signs of steatosis. We then compared fasting plasma glucose, serum insulin and adiponectin, and homoeostasis model assessment of IR (HOMA-IR) and HOMA-beta indices between the groups. RESULTS: There were no significant differences in IR/secretion-associated markers or serum adiponectin. Multivariate analysis demonstrated that the presence of HCV was not an independent predictor of IR. HOMA-IR was strongly correlated with waist circumferences and serum gamma glutamyltransferase in HCV carriers, but not with serum aminotransferase, high sensitivity C-reactive protein, hyaluronic acid or HCV core antigen. CONCLUSIONS: These results suggest that the presence of HCV alone does not affect IR. Coexistence of hepatitis, steatosis and/or fibrosis may be important to the pathogenesis of IR induced by chronic HCV infection. PMID- 18397232 TI - Intracrine signalling of activin A in hepatocytes upregulates connective tissue growth factor (CTGF/CCN2) expression. AB - BACKGROUND/AIMS: Up to now, the effect of activin A on the expression of the important transforming growth factor (TGF)-beta downstream modulator connective tissue growth factor (CTGF) is not known, but might be of relevance for the functional effects of this cytokine on several liver cell types. METHODS: In this study, activin A-dependent CTGF expression in hepatocytes (PC) primed by exogenous activin A and in PC maintained under complete activin-free culture conditions was analysed by Western blots, metabolic labelling, gene silencing, reverse transcriptase-polymerase chain reaction (RT-PCR) and CTGF reporter gene assays. This study was supplemented by immunocytochemical staining of activin A and CTGF in PC of injured liver. RESULTS: Using alkaline phosphatase alpha alkaline phosphatase staining, it is demonstrated that activin A becomes increasingly detectable during the course of CCl(4)-liver damage. Addition of activin A to cultured PC induced CTGF protein expression via phosphorylation of Smad2 and Smad3. This induction can be inhibited by the antagonist follistatin and alpha-activin A antibody respectively. When PC were cultured under serum(i.e. activin A)-free culture conditions, a time-dependent increase of activin expression during the course of the culture was proven by RT-PCR. Silencing of inhibin beta(A) gene expression under serum-free conditions by small interfering RNAs greatly suppressed CTGF synthesis and the phosphorylations of Smad2 and Smad3. However, both the extracellularly acting follistatin and the alpha-activin A antibody could not inhibit spontaneous CTGF expression, which, however, was achieved by the cell-permeable TGF-beta Alk4/Alk5 receptor-kinase-inhibitor SB431542. CONCLUSIONS: In conclusion, the results point to activin A as an inducer of CTGF synthesis in PC. Intracellular activin A contributes to spontaneous CTGF expression in PC independent of exogenous activin A, which is proposed to occur via Alk4/Alk5-receptors. The findings might be important for many actions of activin A on the liver. PMID- 18397233 TI - Outcome and prognostic factors of 391 Japanese patients with primary sclerosing cholangitis. AB - OBJECTIVES: We performed a national survey in 2003, and demonstrated characteristic features of primary sclerosing cholangitis (PSC) patients in Japan. In this study, we aimed to clarify the outcome and prognostic factors of Japanese PSC patients. METHODS: Questionnaires were sent to gastroenterologists in Japan, and 391 patients with PSC were registered and enrolled in the current study. The median follow-up was 5.3 years (range 0.1-20.8 years). The cumulative incidence for survival was analysed using the Kaplan-Meier method. Univariate and multivariate analyses were performed using the Cox-proportional hazards regression model for determining prognostic variables. RESULTS: The estimated median survival of all patients was 13.1 years, with a 5-year survival rate of 74.5%. Thirty-eight patients (9.7%) who underwent liver transplantation (LT) had a 5-year survival rate of 92.0%. Both univariate and multivariate analysis demonstrated that younger age [below 49 years old; odds ratio (OR)=1.76, 1.12 2.76, P=0.0136] and lower total bilirubin (below 3.0 mg/dl; OR=2.50, 1.60-3.89, P or =75 years, compared with those <75 years, the frequencies of angiography and angioplasty procedures were 66.1% versus 87.6% (P < 0.001) and 33.9% versus 53.4% (P < 0.001), respectively; 66% of the elderly with, and without, CHF had angiography. CONCLUSION: The presence of CHF post-MI resulted in lower rates of use of angiography and angioplasty, which was not explained by lower procedure rates in the elderly. As these guideline-recommended procedures may improve survival in patients with CHF post-MI, future strategies should aim to enhance their use. PMID- 18397276 TI - Functional expression of an insect cathepsin B-like counter-defence protein. AB - Insects are capable of readjusting their digestive regimes in response to dietary challenge. Cowpea bruchids (Callosobruchus maculatus) strongly induce C. maculatus cathepsin B-like cysteine protease 1 (CmCatB1) transcripts when fed diet containing a soybean cysteine protease inhibitor soyacystatin N (scN). CmCatB1 shares significant sequence similarity with cathepsin B-like cysteine proteases. In this study, we isolated another cDNA, namely CmCatB2 that encodes a protein sequence otherwise identical to CmCatB1, but lacking a 70-amino-acid internal section. CmCatB1 and CmCatB2 probably resulted from alternate splicing events. Only the CmCatB1 transcript, however, exhibited differential expression in response to dietary scN. Further, this expression was only detectable in larvae, which is the developmental stage associated with food ingestion. The scN activated and developmentally regulated CmCatB1 expression pattern suggests it may have a unique function in insect counter-defence against antinutritional factors. Heterologously expressed recombinant CmCatB1 protein exhibited enzymatic activity in a pH-dependent manner. Activity of the protein was inhibited by both the cysteine protease inhibitor E-64 and the cathepsin B-specific inhibitor CA 074, verifying its cathepsin B-like cysteine protease nature. Interestingly, the enzymatic activity was unaffected by the presence of scN. Together, we have provided functional evidence suggesting that CmCatB1 confers inhibitor insensitive enzymatic activity to cowpea bruchids, which is crucial for insect survival when challenged by dietary protease inhibitors. PMID- 18397278 TI - Molecular cloning and developmental expression of the vitellogenin gene in the endoparasitoid, Pteromalus puparum. AB - A cDNA of the vitellogenin (Vg) protein gene was isolated from the endoparasitoid Pteromalus puparum and characterized. The putative coding sequence was found to be 5634 bp long, encoding 1803 amino acids in a single open reading frame. The chemically determined N-terminal amino acid sequence of vitellin completely matched the deduced amino acid sequence that follows a putative signal peptide of 17 amino acid residues. The Vg mRNA was detected in the fat body of late female pupae, whereas the ovary and male fat body lacked the Vg transcript. The Vg mRNA level in the fat body depended significantly on the developmental stage, reaching the highest level 0 h after eclosion. The haemolymph Vg titre appeared 24 h after the appearance of Vg transcript. A positive correlation between the titre and transcript level of Vg in individual female wasps was found. PMID- 18397277 TI - Cloning and molecular characterization of two invertebrate-type lysozymes from Anopheles gambiae. AB - We sequenced and characterized two novel invertebrate-type lysozymes from the mosquito Anopheles gambiae. Alignment and phylogenetic analysis of these and a number of related insect proteins identified through bioinformatics strategies showed a high degree of conservation of this protein family throughout the Class Insecta. Expression profiles were examined for the two mosquito genes through semiquantitative and real-time PCR analysis. Lys i-1 transcripts were found in adult females in the fat body and Malpighian tubules, whereas Lys i-2 was detected only in fat bodies. Blood-feeding resulted in significantly increased transcript abundance for both genes in the midguts. Neither gene was upregulated following bacterial challenge. PMID- 18397279 TI - EGFR in gastric carcinomas: prognostic significance of protein overexpression and high gene copy number. AB - AIMS: Epidermal growth factor receptor (EGFR) expression has been observed in a variety of solid tumours with the potential of new targeted therapeutic agents. The aim was to evaluate the EGFR status of gastric carcinoma (GC) using immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). METHODS AND RESULTS: The EGFR status was evaluated in GC tissues from 511 patients using IHC and FISH. In addition, the clinicopathological characteristics were examined and the results were compared with the EGFR status. One hundred and forty cases (27.4%) showed EGFR overexpression by IHC. EGFR overexpression was associated with older age (P = 0.001), moderately or poorly differentiated histology (P = 0.001) and higher stage disease (P = 0.046). Sixteen cases (3.1%) showed high polysomy and 12 cases (2.3%) had gene amplification by FISH. The correlation between IHC and FISH results was statistically significant (P < 0.001). The patients with GC who had EGFR overexpression had an unfavourable prognosis and multivariate analysis showed that EGFR overexpression was a possible independent unfavourable prognostic factor. CONCLUSIONS: EGFR overexpression was observed in a subset of cases with GC and was associated with an unfavourable prognosis. It will be important to evaluate EGFR status to interpret future clinical trials properly using EGFR targeted agents. PMID- 18397280 TI - Diffuse large B-cell lymphoma showing an interfollicular pattern of proliferation: a study of the Osaka Lymphoma Study Group. AB - AIMS: Diffuse large B-cell lymphoma (DLBCL) usually proliferates effacing lymph follicles. In occasional cases, tumour cells show an interfollicular pattern of proliferation preserving lymph follicles. The aim was to analyse clinicopathological findings in DLBCL showing an interfollicular pattern of proliferation to determine whether this type of lymphoma is a distinct entity of DLBCL. METHODS AND RESULTS: Clinicopathological findings in 12 cases of DLBCL showing an interfollicular pattern of proliferation [interfollicular group (IF)] were examined and compared with those in 30 cases of DLBCL with ordinary morphology [control group (CG)]. IF showed a significantly lower lactate dehydrogenase level and International Prognostic Index scores than CG (P = 0.023 and P < 0.01, respectively). The frequency of localized disease, clinical stage 1 and 2, in IF was higher than that in CG (P = 0.016). A morphologically polymorphous pattern of proliferation was found in seven of 12 cases (58.3%) in IF, which was higher than that in CG, five (16.7%) of 30 cases (P < 0.01). Clonality analysis with the polymerase chain reaction method revealed that all 11 IF cases examined showed a monoclonal pattern. Immunohistochemically, the majority (11 of 12) of IF cases showed a non-germinal centre B-cell phenotype and the frequency was higher than that in CG (P = 0.021). CONCLUSION: Diffuse large B cell lymphoma with an interfollicular pattern of proliferation shows distinct clinical and pathological findings from ordinary DLBCL. PMID- 18397282 TI - Classifying thymomas. PMID- 18397283 TI - Haemophilic knee arthropathy with extension contracture. PMID- 18397281 TI - Impact of immunohistochemical markers, CK5/6 and E-cadherin on diagnostic agreement in non-invasive proliferative breast lesions. AB - AIMS: To assess the impact of cytokeratin (CK) 5/6 and E-cadherin immunohistochemistry on diagnostic agreement of non-invasive proliferative breast lesions. METHODS AND RESULTS: Twenty pathologists classified 105 cases of non invasive proliferative breast lesions into 10 diagnostic categories. One haematoxylin and eosin (H&E) slide of each case was analysed on a first round and one H&E slide with corresponding CK5/6 and E-cadherin immunohistochemistry was analysed on a second round. Interobserver reproducibility for category-specific and management-specific lesions was measured on each round. CK5/6 and E-cadherin had little impact on diagnostic agreement, which remained moderate between the first and second rounds (overall kappa coefficients of 0.47 and 0.53, respectively, P = NS). Levels of agreement slightly improved for lesions with specific CK5/6 and E-cadherin immunoprofiles (usual ductal hyperplasia, atypical ductal hyperplasia, atypical lobular hyperplasia, lobular carcinoma in situ, non high-grade ductal carcinoma in situ), but the differences observed were not statistically significant. However, diagnostic agreement improved when lesions were grouped according to their management category (overall kappa coefficients of 0.58 and 0.66 in the first and second rounds, respectively). CONCLUSIONS: CK5/6 and E-cadherin immunohistochemistry has little impact on interobserver reproducibility for non-invasive breast lesions. Diagnostic agreement can, however, be improved by grouping lesions in management categories. PMID- 18397284 TI - Retrograde intramedullary nailing of supracondylar femoral fractures in haemophilic patients. PMID- 18397285 TI - L1503R is a member of group I mutation and has dominant-negative effect on secretion of full-length VWF multimers: an analysis of two patients with type 2A von Willebrand disease. AB - Type 2A von Willebrand disease (VWD) is characterized by decreased platelet dependent function of von Willebrand factor (VWF); this in turn is associated with an absence of high-molecular-weight multimers. Sequence analysis of the VWF gene from two unrelated type 2A VWD patients showed an identical, novel, heterozygous T-->G transversion at nucleotide 4508, resulting in the substitution of L1503R in the VWF A2 domain. This substitution, which was not found in 60 unrelated normal individuals, was introduced into a full-length VWF cDNA and subsequently expressed in 293T cells. Only trace amount of the mutant VWF protein was secreted but most of the same was retained in 293T cells. Co-transfection experiment of both wild-type and mutant plasmids indicated the dominant-negative mechanism of disease development; as more of mutant DNA was transfected, VWF secretion was impaired in the media, whereas more of VWF was stored in the cell lysates. Molecular dynamic simulations of structural changes induced by L1503R indicated that the mean value of all-atom root-mean-squared-deviation was shifted from those with wild type or another mutation L1503Q that has been reported to be a group II mutation, which is susceptible to ADAMTS13 proteolysis. Protein instability of L1503R may be responsible for its intracellular retention and perhaps the larger VWF multimers, containing more mutant VWF subunits, are likely to be mal-processed and retained within the cell. PMID- 18397286 TI - Corpus luteal hemorrhage: an unusual manifestation of congenital factor XIII deficiency. PMID- 18397287 TI - Heterogeneity of large clostridial toxins: importance of Clostridium difficile toxinotypes. AB - Clostridium difficile toxinotypes are groups of strains defined by changes in the PaLoc region encoding two main virulence factors: toxins TcdA and TcdB. Currently, 24 variant toxinotypes (I-XXIV) are known, in addition to toxinotype 0 strains, which contain a PaLoc identical to the reference strain VPI 10463. Variant toxinotypes can also differ from toxinotype 0 strains in their toxin production pattern. The most-studied variant strains are TcdA-, TcdB+ (A-B+) strains and binary toxin CDT-producing strains. Variations in toxin genes are also conserved on the protein level and variant toxins can differ in size, antibody reactivity, pattern of intracellular targets (small GTPases) and consequently in their effects on the cell. Toxinotypes do not correlate with particular forms of disease or patient populations, but some toxinotypes (IIIb and VIII) are currently associated with disease of increased severity and outbreaks worldwide. Variant toxinotypes are very common in animal hosts and can represent from 40% to 100% of all isolates. Among human isolates, variant toxinotypes usually represent up to 10% of strains but their prevalence is increasing. PMID- 18397288 TI - L-Proline uptake in Crithidia deanei is influenced by its endosymbiont bacterium. AB - Crithidia deanei, a monoxenic trypanosomatid, presents an endosymbiotic bacterium in its cytoplasm. Both the protozoan and the bacterium maintain intensive metabolic exchange, resulting in an interesting model to study the coevolution of metabolisms. The relevance of l-proline for the growth of C. deanei and its transport into these cells was studied. Both the endosymbiont-containing (wild) and the endosymbiont-free protozoa (aposymbiont or cured) strains, when grown in medium supplemented with l-proline, reached higher cell densities than those grown in unsupplemented media. We biochemically characterized the uptake of l proline in both the wild (K(m)=0.153+/-0.022 mM, V(max)=0.239+/-0.011 nmol min( 1) per 4 x 10(7) cells) and the aposymbiont strains (K(m)=0.177+/-0.049 mM, V(max)=0.132+/-0.012 nmol min(-1) per 4 x 10(7) cells). These data suggest a single type of proline transporter whose activity is upregulated by the presence of the symbiotic bacterium. Proline transport was further characterized and was found to be insensitive to the extracellular concentration of Na+, but sensitive to K+ and pH. The abolition of proline uptake by respiratory chain inhibitors and valinomycin indicates that the proline transport in C. deanei is dependent on the plasma membrane K+ gradient. PMID- 18397289 TI - The type III secretion system-dependent repression of NF-kappaB activation to the intracellular growth of Edwardsiella tarda in human epithelial cells. AB - Edwardsiella tarda is a pathogen with a broad host range infecting animals and humans. We have reported recently that the type III secretion system (TTSS) is essential for intracellular replication of the bacterium in murine macrophages. The present study shows that the TTSS is also needed for intracellular growth of the bacterium in human epithelial cells (HEp-2). However, different from the previous microarray analyses on murine macrophages, upregulation of the mRNA expression level of NF-kappaB target genes was not detected in the infected HEp-2 cells. The wild-type E. tarda, but not its TTSS mutant, actually repressed the tumor necrosis factor alpha-dependent NF-kappaB activation in an NF-kappaB reporter gene assay. These results suggest TTSS-dependent repression of the NF kappaB activation in HEp-2 cells infected with E. tarda. PMID- 18397290 TI - Nicotinamide inhibits Plasmodium falciparum Sir2 activity in vitro and parasite growth. AB - Plasmodium falciparum sirtuin, PfSir2, contains histone deacetylase (HDAC) activity that may be central to the regulation of virulence gene expression in the parasites. Although a few reports have been published recently regarding in vitro and in vivo function of PfSir2, expression of the endogenous protein (c. 30 kDa) has not been shown yet. Here we report the presence of PfSir2 in the parasite at the protein level by specific antibodies. HDAC activity of PfSir2 can be inhibited by nicotinamide, a product of sirtuin reaction. Surprisingly, we find that nicotinamide also delays parasite growth significantly in culture. These findings further our knowledge on PfSir2 and raise the possibility of using an inexpensive agent like nicotinamide as an antimalarial in combination with other antiparasitic drugs. PMID- 18397291 TI - Quorum sensing in metal tolerance of Acinetobacter junii BB1A is associated with biofilm production. AB - Acinetobacter junii strain BB1A, a novel metal-tolerant bacterium, produced biofilm in the presence of added ions such as Ni(2+), AsO(2)(-), Cd(2+) and Hg(2+) on surfaces such as glass and polystyrene. Generation of a metal-sensitive and adhesion-deficient mutant by transposition of Tn5-mob in the A. junii genome has putatively confirmed the association of metal tolerance with the production of biofilm. The requirement of a critical cell density for biofilm formation and presence of acyl-homoserine lactone-like autoinducer molecules in the cell-free supernatant indicated the phenomenon of quorum sensing. Addition of a natural quorum-sensing inhibitor (garlic extract) or synthetic quorum-sensing inhibitor (4-nitro-pyridine oxide) significantly inhibited cell growth and biofilm formation in the presence of metal/metalloid ions. PMID- 18397292 TI - Anticonvulsive and antiepileptogenic effects of levetiracetam in the audiogenic kindling model. AB - PURPOSE: To study anticonvulsive and antiepileptogenic effects of singe levetiracetam (LEV) administration in the model of audiogenic kindling. METHODS: Rats of Krushinsky-Molodkina (KM) strain genetically susceptible to severe audiogenic seizures received one intraperitoneal injection of saline, low (6 mg/kg) or high (50 mg/kg) dose of LEV before or after audiogenic kindling. One hour postinjection, an audiogenic seizure was induced to assess anticonvulsive effect of LEV in nonkindled and kindled rats. To examine antiepileptogenic activity of LEV, nonkindled rats injected with the drug or saline were kindled with repeated sound stimulations. Audiogenic kindling development manifested in an appearance and progressive prolongation of an additional seizure phase, post tonic-clonus. The latency and duration of audiogenic seizures and the duration of every seizure phase (running, tonic, post-tonic-clonic) were measured. RESULTS: One hour posttreatment, LEV dose-dependently lengthened the latency and reduced the duration of audiogenic seizures in both nonkindled and kindled rats. The seizure shortening resulted from selective suppression of tonic and kindled post tonic-clonic phases. The dose of 50 mg/kg completely blocked tonic and clonic convulsions 1 h postinjection. The anticonvulsive effect of LEV was more pronounced in kindled than in nonkindled rats. Single LEV injection in the dose of 50 mg/kg prior audiogenic kindling significantly suppressed subsequent kindling progression indicating profound antiepileptogenic potency of the drug. CONCLUSIONS: The present study shows that LEV exerts both short-lasting anticonvulsive effect on audiogenic seizures and very long-lasting antiepileptogenic effect on audiogenic kindling. Remarkably, a single injection of LEV is enough to significantly suppress kindling progression in KM rats. PMID- 18397293 TI - Decreases in HCN mRNA expression in the hippocampus after kindling and status epilepticus in adult rats. AB - PURPOSE: Studies in animal models and patients have implicated changes in hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN) expression in the pathogenesis of temporal lobe epilepsy (TLE). However, the nature of HCN changes during the epileptogenic process and their commonality across different TLE models is unknown. Here HCN1 and HCN2 mRNA expression was quantitatively measured at different time points during epileptogenesis in two distinct animal models of TLE; the kainic acid (KA)-induced status epilepticus (SE) and amygdala kindling models. METHODS: Hippocampal subregions (CA1, CA3, and dentate gyrus [DG]) and entorhinal cortex were dissected at different time points. For KA-induced SE animals this was 24 h, 7 days (preepileptic), and 6 weeks (epileptic) post status. For amygdala kindling animals this was 2 weeks after reaching either "partially kindled" (one class II/III seizure) or "fully kindled" (five class V seizures) states. Quantification of regional hippocampal neuronal loss in the KA-treated animals was done using NeuN immunofluorescence and confocal microscopy. RESULTS: HCN mRNA levels decreased in an isoform and region specific manner at all time points after KA-induced SE. The decrease in neuronal number could not account for all reductions in HCN mRNA levels post-KA insult, implicating transcriptional changes. A reduction in HCN2 mRNA levels was also observed in fully kindled animals in the CA3 region. CONCLUSIONS: A reduction in HCN mRNA levels is present in two different models of TLE. This supports the case that a reduction in HCN channel expression is an accompaniment of epileptogenesis in different adult models of TLE. PMID- 18397294 TI - Abnormalities in diffusion tensor imaging of the uncinate fasciculus relate to reduced memory in temporal lobe epilepsy. AB - AIMS: To use Diffusion Tensor Imaging (DTI) to explore structural integrity and connectivity of the uncinate fasciculus (UF) in patients with temporal lobe epilepsy (TLE) and its relationship to memory performance. METHODS: DTI and UF reconstruction were performed in 28 patients with TLE (18 left, 10 right) and 10 normal controls. Differences between left and right UF fractional anisotropy (FA) and apparent diffusion coefficient (ADC) values and correlations between DTI measures and memory scores in the TLE groups were computed. RESULTS: In controls, FA was higher in the left than right UF (p < 0.01). In left TLE, FA values were lower and ADC values higher than controls in the left UF and ADC values were higher in the right UF (all p < 0.05). In right TLE, ADCs were higher in the left and right UF compared to controls, and FA was reduced in the left UF (all p < 0.05). In left TLE, ADCs in the left UF were negatively correlated with Auditory Immediate (p < 0.05) and Delayed Memory (p < 0.01). Visual Delayed Memory was positively correlated with reduced FA in the ROI of the right UF and increased radial diffusivities (p < 0.05). No significant correlations were found in right TLE. Thus, DTI values correlated with memory scores in the expected direction in patients with left TLE. CONCLUSIONS: Abnormal diffusion measures in the UF ipsilateral to the epileptogenic zone suggest that integrity of the UF is related to memory performance in patients with left TLE. Larger sample sizes are needed to evaluate structure-function correlations further. PMID- 18397296 TI - Effects of high-frequency stimulation on epileptiform activity in vitro: ON/OFF control paradigm. AB - PURPOSE: To determine the effects of high-frequency electrical stimulation on electrographic seizure activity during and after stimulation (ON-effect and OFF effect). METHODS: The modulation and suppression of epileptiform activity during (ON-effect) and after (OFF-effect) high-frequency electrical stimulation was investigated using the high-K(+) and picrotoxin hippocampal slice epilepsy models. Uniform sinusoidal fields (50 Hz) were applied with various intensity levels for 1 min across brain slices. Extracellular and intracellular activity were monitored during and after stimulation. RESULTS: The ON-effects of high frequency stimulation were highly variable across individual slices and models; ON-effects included modulation of activity, pacing, partial suppression, or activity resembling spreading-depression. On average, epileptic activity, measured as power in the extracellular fields, increased significantly during stimulation. Following the termination of electrical stimulation, a robust poststimulation suppression period was observed. This OFF suppression was observed even at relatively moderate stimulation intensities. The duration of OFF suppression increased with stimulation intensity, independent of ON-effects. Antagonism of GABA(A)function did not directly effect OFF suppression duration. CONCLUSIONS: The present results suggest that "rational" seizure control protocols using intermittent high-frequency electrical stimulation should control for both ON and OFF effects. PMID- 18397295 TI - Granule cell dispersion develops without neurogenesis and does not fully depend on astroglial cell generation in a mouse model of temporal lobe epilepsy. AB - PURPOSE: Granule cell dispersion (GCD) appears as a characteristic morphological feature of the mesial temporal lobe epilepsy (MTLE). It has been suggested that this phenomenon could be due to an increased neurogenesis in the dentate gyrus. However, this hypothesis is still debated and recent clinical and experimental studies have shown that neurogenesis is rather decreased in MTLE. To further determine the role of neural and astroglial cell generation in GCD we examined the consequences of aging and irradiation, which are known to reduce progenitor cells, in a mouse model of MTLE induced by intrahippocampal kainate (KA) injection. METHODS: We injected KA in hippocampus of three different types of mice; (1) young adult, (2) aged, and (3) irradiated mice. Newly generated cells were labeled by Bromodeoxyuridine (BrdU) and were characterized by immunohistochemistry. The extent of GCD was compared among the three animal groups. RESULTS: In young adult mice, BrdU-labeled neurons as well as doublecortin- and NeuroD-positive cells decreased progressively after KA injection whereas BrdU-labeled astrocytes and microglias increased. In aged and irradiated mice, where basal neurogenesis was already strongly reduced, GCD developed after KA injection to the same extent as in young adult mice. However, augmentation of the BrdU-labeled astrocytes after KA was less than 40% in irradiated mice in comparison to young and aged mice. CONCLUSIONS: Our data show that GCD occurs without neurogenesis. Furthermore GCD developed regardless of the degree of astroglial cell proliferation, suggesting that neural stem cell generation is not crucial for GCD. PMID- 18397297 TI - Wind turbines, flicker, and photosensitive epilepsy: characterizing the flashing that may precipitate seizures and optimizing guidelines to prevent them. AB - Wind turbines are known to produce shadow flicker by interruption of sunlight by the turbine blades. Known parameters of the seizure provoking effect of flicker, i.e., contrast, frequency, mark-space ratio, retinal area stimulated and percentage of visual cortex involved were applied to wind turbine features. The proportion of patients affected by viewing wind turbines expressed as distance in multiples of the hub height of the turbine showed that seizure risk does not decrease significantly until the distance exceeds 100 times the hub height. Since risk does not diminish with viewing distance, flash frequency is therefore the critical factor and should be kept to a maximum of three per second, i.e., sixty revolutions per minute for a three-bladed turbine. On wind farms the shadows cast by one turbine on another should not be viewable by the public if the cumulative flash rate exceeds three per second. Turbine blades should not be reflective. PMID- 18397298 TI - Prolonged neurophysiologic effects of levetiracetam after oral administration in humans. AB - PURPOSE: To determine whether neurophysiological effects of levetiracetam (LEV) outlast its serum half-life of approximately 7 h. Demonstration of prolonged effects would help to explain the efficacy of LEV at conventional dosing intervals that are longer than the serum half-life. METHODS: Following an oral dose of LEV 3 g in 12 normal volunteers, we compared transcranial magnetic stimulation (TMS) measures of motor threshold (MT) and recruitment with LEV serum levels and subjective ratings of toxicity over 48 h. Subjects used a two dimensional visual-analog scale to estimate the time course of any side effects. RESULTS: LEV serum levels and subjective toxicity both peaked around 1 h after oral administration. MT elevation was delayed in comparison to peak serum levels and subjective toxicity. MT was maximally elevated at 6-9 h, and recruitment maximally reduced at 0.6-9 h. Changes in both measures had recovered by approximately 50% at 24 h. Despite the time difference between toxicity and TMS changes, toxicity estimates correlated with the maximum increase in MT. CONCLUSION: There is a substantial time lag between LEV serum levels and TMS measures of neuronal effects, and a similar temporal dissociation between subjective toxicity and maximum TMS change. The time course of neurophysiological effects, as measured by TMS, may help to explain the sustained clinical efficacy of LEV despite a short peripheral half-life. PMID- 18397299 TI - Antiepileptic drugs--best practice guidelines for therapeutic drug monitoring: a position paper by the subcommission on therapeutic drug monitoring, ILAE Commission on Therapeutic Strategies. AB - Although no randomized studies have demonstrated a positive impact of therapeutic drug monitoring (TDM) on clinical outcome in epilepsy, evidence from nonrandomized studies and everyday clinical experience does indicate that measuring serum concentrations of old and new generation antiepileptic drugs (AEDs) can have a valuable role in guiding patient management provided that concentrations are measured with a clear indication and are interpreted critically, taking into account the whole clinical context. Situations in which AED measurements are most likely to be of benefit include (1) when a person has attained the desired clinical outcome, to establish an individual therapeutic concentration which can be used at subsequent times to assess potential causes for a change in drug response; (2) as an aid in the diagnosis of clinical toxicity; (3) to assess compliance, particularly in patients with uncontrolled seizures or breakthrough seizures; (4) to guide dosage adjustment in situations associated with increased pharmacokinetic variability (e.g., children, the elderly, patients with associated diseases, drug formulation changes); (5) when a potentially important pharmacokinetic change is anticipated (e.g., in pregnancy, or when an interacting drug is added or removed); (6) to guide dose adjustments for AEDs with dose-dependent pharmacokinetics, particularly phenytoin. PMID- 18397300 TI - Spatial and temporal variability in epilithic biofilm bacterial communities along an upland river gradient. AB - Riverine biofilms remain one of the least-studied habitats despite the significant increase in the examination of aquatic microbial communities in recent years. In this study, the dynamics of epilithic biofilm communities native on rocks from a low-order upland stream were examined over a period of 3 years. Spatial and temporal variations in bacterial communities were assessed using terminal restriction fragment length polymorphism, based on analysis of the 16S rRNA gene. In total, 108 epilithic biofilm samples were analysed and 170 different ribotypes were detected. A strong temporal gradient in ribotype composition was noticed, especially between samples collected in 2001 and those collected in 2002 and 2003, most likely reflecting interannual differences in weather conditions, such as temperature. A spatial gradient in ribotype composition, from upstream sites to the low-lying sites, was also evident and interpreted as an environmental variation gradient along the river course. Distinct biofilm communities consistently occurred at the first site along the river, which was significantly correlated to low pH. Temporal factors explained the highest degree of variation within the epilithic biofilms. Recurrent blooms of certain bacteria were noted within the system. Phylogenetic relationships of bacteria at one point in the river were determined using a cloning and sequencing approach, with Alphaproteobacteria dominating the community, followed by Cyanobacteria, Bacteroidetes and Betaproteobacteria. PMID- 18397302 TI - Regulatory region polymorphisms of vitamin D receptor gene in pulmonary tuberculosis patients and normal healthy subjects of south India. AB - Vitamin D receptor (VDR) gene variants are associated with differential susceptibility or resistance to tuberculosis in different ethnic groups. We investigated the polymorphisms in the 5' regulatory region of VDR gene in 206 normal healthy subjects and 166 patients with pulmonary tuberculosis from south India. Cdx-2 polymorphism was studied by polymerase chain reaction (PCR) with allele-specific primers, while genotyping of A1012G was done by PCR-based restriction fragment length polymorphism. A significantly decreased frequency of Cdx-2 G allele (P = 0.016) and G/G genotype (P = 0.010) and an increased frequency of A-A haplotype (A allele of Cdx-2 and A allele of A1012G) (P = 0.015) were observed in patients compared to controls. The study suggests that Cdx-2 G/G genotype may be associated with protection and A-A haplotype with susceptibility to tuberculosis. PMID- 18397301 TI - Human MHC architecture and evolution: implications for disease association studies. AB - Major histocompatibility complex (MHC) variation is a key determinant of susceptibility and resistance to a large number of infectious, autoimmune and other diseases. Identification of the MHC variants conferring susceptibility to disease is problematic, due to high levels of variation and linkage disequilibrium. Recent cataloguing and analysis of variation over the complete MHC has facilitated localization of susceptibility loci for autoimmune diseases, and provided insight into the MHC's evolution. This review considers how the unusual genetic characteristics of the MHC impact on strategies to identify variants causing, or contributing to, disease phenotypes. It also considers the MHC in relation to novel mechanisms influencing gene function and regulation, such as epistasis, epigenetics and microRNAs. These developments, along with recent technological advances, shed light on genetic association in complex disease. PMID- 18397303 TI - Interleukin-10 promoter polymorphisms in patients with systemic lupus erythematosus from the Canary Islands. AB - The purpose of this study was to examine whether several allelic variants in the polymorphic interleukin (IL)-10 promoter region were related with an increased risk of developing systemic lupus erythematosus (SLE) in Spanish patients from Canary Islands. Microsatellites (MS) at positions -4000 and -1200 (IL10R and IL10G, respectively) and single nucleotide polymorphisms (SNPs) (MS) at positions -1082G/A, -819C/T and -592C/A of the IL-10 promoter were analysed in patients with SLE and healthy controls from Canary Islands (Spain). We found that SNPs but not MS were associated with SLE. The GCC haplotype frequency was significantly higher in SLE patients (0.43) than in healthy donors (0.33) [P = 0.02; OR = 1.50 (95% CI = 1.06-2.14)], whereas the ACC haplotype was less represented in patients (0.28 vs. 0.37) [P = 0.02; OR = 0.64 (95% CI = 0.44-0.92)]. To assess the functional role of genotypes, serum IL-10 levels from patients and controls were quantified by ELISA. Also, the lipopolysaccharide-induced IL-10 secretion by monocytes from healthy controls was evaluated in vitro. Serum IL-10 levels were higher in patients [median (interquartile range) = 2.8 pg/mL (1.8-4.2)] than in controls [0.9 pg/mL (0-3.5)] (P = 0.02), but no association was observed between serum IL-10 levels or lipopolysaccharide-induced IL-10 secretion and the IL-10 promoter haplotypes. These data suggest that the IL-10 promoter haplotype that produces higher levels of cytokine is associated with SLE in patients from Canary Islands. PMID- 18397304 TI - Takotsubo cardiomyopathy associated with status epilepticus. PMID- 18397305 TI - Effects of induced stress on experimental pain sensitivity in chronic tension type headache sufferers. AB - BACKGROUND AND PURPOSE: It has been proposed that stress may contribute to chronic tension-type headache (CTH) through hyperalgesic effects on already sensitized pain pathways in CTH sufferers. This hypothesis could be partially tested by examining effects of stress on mechanical and thermal pain sensitivity in CTH sufferers. Such examinations have not been reported to date. MATERIALS AND METHODS: In the present study, we measured cephalic and extra-cephalic pressure [pressure-pain threshold (PPT)] and cold-pain thresholds (CPT) in CTH sufferers (n = 8 females, n = 8 males) and healthy control subjects (n = 8 males, n = 7 females) recruited from the general population before and after exposure to a 15 min stressful mental task. RESULTS: Results indicated that PPT's at head and hand were lower in the CTH compared with control group both before and after task exposure. PPT's and CPT's decreased from pre- to post-task in both groups, with a significantly greater pre- to post-task reduction in cephalic PPT in the CTH compared with control group. Subjective stress increased from pre- to post-task in both groups and did not differ between groups. In the CTH group, stress reactivity was negatively correlated with PPT's, whilst absolute stress levels were positively correlated with pre- to post-task decrease in PPT's. CONCLUSIONS: The main finding is an enhanced hyperalgesic effect of stress on cephalic pressure-pain sensitivity in the CTH sufferers compared with the healthy controls. The results support the hypothesis that stress may contribute to CTH through hyperalgesic effects on already sensitized pain pathways in CTH sufferers. PMID- 18397306 TI - Bacterial ectosymbionts and virulence silencing in a Fusarium oxysporum strain. AB - In the present article we have ascertained the presence of a consortium of ectosymbiotic bacteria belonging to Serratia, Achromobacter, Bacillus and Stenotrophomonas genera associated to the mycelium of the antagonistic Fusarium oxysporum MSA 35 [wild-type (WT) strain]. Morphological characterization carried out on the WT strain, on the F. oxysporum MSA 35 without ectosymbionts [cured (CU) strain] and on the pathogenic F. oxysporum f.sp. lactucae (Fuslat 10) showed that the ectosymbionts, present only in the WT strain, caused a depleted production of micro conidia and aerial hyphae, and a change in shape and dimension of the latter. Virulence tests showed that the cured Fusarium was a pathogenic strain and, as shown by polymerase chain reaction and microscope analysis, pathogenicity was correlated with the capability of the cured hyphae of penetrating lettuce roots. Accordingly, the hyphae of the WT strain were impaired in entering the plant roots. Typing experiments provided evidence that both CU and WT strains belong to F. oxysporum f.sp. lactucae. This implies that the antagonistic effect of WT Fusarium is not a fungal trait, but it is due to the interaction with the ectosymbiotic bacteria. Expression analysis showed that fmk1, chsV and pl1 genes involved in F. oxysporum pathogenicity are not expressed in the WT strain whereas they are expressed in the cured fungus. These results, together with the hyphal characteristics, suggest that the inability of WT strain to penetrate the plant roots could be due to alterations in the expression profile of cell wall-degrading enzymes. In conclusion, we demonstrated a modulation of F. oxysporum gene expression in response to the interaction with the ectosymbiotic bacteria. Preliminary researches indicated that the presence of bacteria attached to the hyphae of antagonistic F. oxysporum is not an isolated phenomenon. Further investigations are necessary to better understand the rule and the diffusion of ectosymbiotic bacteria among antagonistic Fusarium. PMID- 18397307 TI - Changes in the photosynthetic reaction centre II in the diatom Phaeodactylum tricornutum result in non-photochemical fluorescence quenching. AB - Diatoms are an important group of primary producers in the aquatic environment. They are able to acclimate to fast changes in the light intensity by various mechanisms including a rise in non-photochemical fluorescence quenching (NPQ). The latter has been attributed to the xanthophyll cycle (XC) following activation of diadinoxanthin de-epoxidase by the acidification of the thylakoid lumen. To examine whether fluorescence quenching in the diatom Phaeodactylum tricornutum depends on the DeltapH generated by the photosynthetic electron transport, we arrested the latter by 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU). This treatment hardly affected the NPQ or XC, even when methylviologen was present. Dissipation of the DeltapH by 2,4-dinitrophenol inhibited the XC but did not alter NPQ. Similar results, i.e. inhibition of the XC but normal fluorescence quenching, were observed when the experiments were performed at 3 degrees C. Measurements of thermoluminescence showed that excess light treatment caused a marked decline in the signals obtained as a result of recombination of Q(B) (-) with the S(3) state of the Mn cluster; this was also observed in cells treated with DCMU (recombination of Q(A) (-) with S(2)). Light treatment also diminished the Q(A) (-) re-oxidation signals. The data suggest that changes in PSII core centre itself due to exposure to excess light conditions play an important part in the acclimation of P. tricornutum to the changing light conditions. PMID- 18397308 TI - The hyperthermophilic anaerobe Thermotoga Maritima is able to cope with limited amount of oxygen: insights into its defence strategies. AB - Thermotoga maritima, an anaerobic hyperthermophilic bacterium, was found able to grow in the presence of low concentrations of oxygen of up to 0.5% (v/v). Differential proteomics and transcripts analysis by qRT-PCR were used to identify the defence strategies used by T. maritima to protect itself against oxygen. A flavoprotein, homologous to rubredoxin oxygen oxidoreductase was found to be overproduced when cells were cultured in oxidative conditions. The recombinant protein, produced in Escherichia coli, exhibited an oxygen reductase activity, which could account for the observed decrease in oxygen concentration during growth. The gene encoding this oxygen reductase belongs to a multicistronic unit that includes genes encoding proteins involved in exopolysaccharide biosynthesis, which may be related to a biofilm formation induced by the presence of oxygen. Enzymes involved in reactive oxygen species detoxification, iron-sulfur centre synthesis/repair and the cysteine biosynthesis pathway were also overproduced. All these enzymatic systems together contribute to the defence strategy of T. maritima against oxygen. Because of the position of T. maritima in deep branches of the phylogenetic tree, we suggest that these strategies can be considered as ancestral mechanisms first developed by anaerobic microorganisms on the early Earth to protect themselves against primary abiotic or biotic oxygen production. PMID- 18397309 TI - Does fossil pigment and DNA data from Mediterranean sediments invalidate the use of green sulfur bacterial pigments and their diagenetic derivatives as proxies for the assessment of past photic zone euxinia? PMID- 18397310 TI - Using a genetic algorithm to drive a microbial ecosystem in a desirable direction. AB - The functioning of natural microbial ecosystems is influenced by various biotic and abiotic conditions. The careful experimental manipulation of environmental conditions can drive microbial ecosystems toward exhibiting desirable types of functionality. Such manipulations can be systematically approached by viewing them as a combinatorial optimization problem, in which the optimal configuration of environmental conditions is sought. Such an effort requires a sound optimization technique. Genetic algorithms are a class of optimization methods that should be suitable for such a task because they can deal with multiple interacting variables and with experimental noise and because they do not require an intricate understanding or modelling of the ecosystem of interest. We propose the use of genetic algorithms to drive undefined microbial ecosystems in desirable directions by combinatorially optimizing sets of environmental conditions. We tested this approach in a model system where the microbial ecosystem of a human saliva sample was manipulated in successive steps to display increasing amounts of azo dye decoloration. The results of our experiments indicated that a genetic algorithm was capable of optimizing ecosystem function by manipulating the presence or absence of a set of 10 chemical supplements. Genetic algorithms hold promise for use as a tool in environmental microbiology for the efficient control of the functioning of natural and undefined microbial ecosystems. PMID- 18397311 TI - Sources for sedimentary bacteriohopanepolyols as revealed by 16S rDNA stratigraphy. AB - Bacteriohopanoids are widespread lipid biomarkers in the sedimentary record. Many aerobic and anaerobic bacteria are potential sources of these lipids which sometimes complicates the use of these biomarkers as proxies for ecological and environmental changes. Therefore, we applied preserved 16S ribosomal RNA genes to identify likely Holocene biological sources of bacteriohopanepolyols (BHPs) in the sulfidic sediments of the permanently stratified postglacial Ace Lake, Antarctica. A suite of intact BHPs were identified, which revealed a variety of structural forms whose composition differed through the sediment core reflecting changes in bacterial populations induced by large changes in lake salinity. Stable isotopic compositions of the hopanols formed from periodic acid-cleaved BHPs, showed that some were substantially depleted in (13)C, indicative of their methanotrophic origin. Using sensitive molecular tools, we found that Type I and II methanotrophic bacteria (respectively Methylomonas and Methylocystis) were unique to the oldest lacustrine sediments (> 9400 years BP), but quantification of fossil DNA revealed that the Type I methanotrophs, including methanotrophs related to methanotrophic gill symbionts of deep-sea cold-seep mussels, were the main precursors of the 35-amino BHPs (i.e. aminopentol, -tetrol and -triols). After isolation of the lake approximately 3000 years ago, one Type I methanotroph of the 'methanotrophic gill symbionts cluster' remained the most obvious source of aminotetrol and -triol. We, furthermore, identified a Synechococcus phylotype related to pelagic freshwater strains in the oldest lacustrine sediments as a putative source of 2-methylbacteriohopanetetrol (2-Me BHT). This combined application of advanced geochemical and paleogenomical tools further refined our knowledge about Holocene biogeochemical processes in Ace Lake. PMID- 18397312 TI - Stoichiometry of endothermy: shifting the quest from nitrogen to carbon. AB - For many animals, notably herbivores, plants are often an inadequate food source given the low content of protein and high content of C-rich material. This conception is mainly based on studies on ectotherms. The validity of this conception for endotherms is unclear given their much higher carbon requirements for maintenance energy metabolism than ectotherms. Applying stoichiometric principles, we hypothesized that endotherms can cope with diets with much higher (metabolizable) carbon to nitrogen ratios than ectotherms. Using empirical data on birds, eutherian mammals, marsupials and reptiles, we compiled and compared measurements and allometric equations for energy metabolism as well as nitrogen requirements. Our analysis supports our hypothesis that plants, and especially their leaves, are generally sufficiently rich in nitrogen to fulfil protein demands in endotherms, at least during maintenance conditions, but less so in ectotherms. This has important implications with respect to community functioning and the evolution of endothermy. PMID- 18397313 TI - Reticular erythema with focal mid-dermal elastophagocytosis (REMDE). AB - SUMMARY: A 57-year-old man presented with a persistent arcuate partly reticular erythema of the upper chest. Histopathological examination revealed an upper and mid-dermal perivascular and interstitial dermatitis with macrophages which had engulfed elastic fibers. Additionally a focal dermal elastolysis was observed. The characteristic clinicopathological constellation has hitherto been reported in only 3 cases, including our case. Presumably, the disease belongs to a spectrum of clinical manifestation of mid-dermal elastolysis. It is suggested to gather future cases under the provisional diagnosis ''reticular erythema with focal mid-dermal elastophagocytosis" (REMDE) to gain more insights into this rare disease. PMID- 18397314 TI - Peas, beans, and the Pythagorean theorem - the relevance of glucose-6-phosphate dehydrogenase deficiency in dermatology. AB - Glucose-6-phosphate (G6PD) deficiency is a common disease characterized by acute hemolysis induced by oxidative stress. More than 400 million subjects throughout the world carry the hereditary enzyme defect with the highest prevalences in Africa, Asia, and the Mediterranean region. In individuals affected by the erythrocytic enzymatic disorder, besides infectious diseases and diet, acute hemolytic crisis can be triggered by numerous drugs frequently used for the treatment of dermatoses. Taking into account the increasing number of immigrants from geographic regions with high prevalences of G6PD deficiency, dermatologists should be alert to the presence of disease. PMID- 18397315 TI - Screening of glucose/insulin metabolic alterations in men with multiple skin tags on the neck. AB - SUMMARY: Multiple skin tags appear associated with abnormalities in glucose/insulin metabolism. Clinical and metabolic glucose/insulin characteristics of men with multiple (8 or more) skin tags on the neck were compared with a control group with few or none. Both groups were divided in two subgroups according to normal or abnormal laboratory findings. In the study subgroup with normal laboratory findings the number of skin tags varied from 8 33, whereas in those with abnormal laboratory findings the range was 9-65. Eight or more skin tags were related with statistically significant laboratory glucose/insulin abnormalities: basal hyperinsulinemia (p<0.002), postprandial hyperinsulinemia (p<0.003), and postprandial hyperglycemia (p<0.01). In the multiple skin tag group 77 % had diverse laboratory abnormalities, including insulin resistance, basal hyperinsulinemia, postprandial hyperinsulinemia, glucose intolerance or type 2 diabetes, in contrast with the control group, where only 33 % showed laboratory abnormalities. One-third of the study group had acanthosis nigricans. Only 15 % of patients with metabolic abnormalities did not show any cutaneous expression of glucose/insulin alterations (9 or more skin tags on the neck, acanthosis nigricans, or waist circumference greater than 95 cm). Multiple skin tags were more sensitive than acanthosis nigricans in identifying those with alterations in the glucose/insulin metabolism (77 vs. 32 % respectively), although less specific (68 vs.100%). Multiple skin tags should raise suspicion of insulin resistance or hyperinsulinemia. PMID- 18397316 TI - Interaction with model membranes and pore formation by human stefin B: studying the native and prefibrillar states. AB - Human stefin B, from the family of cystatins, is used as a model amyloidogenic protein in studies of the mechanism of amyloid fibril formation and related cytotoxicity. Interaction of the protein's prefibrillar oligomers/aggregates with predominantly acidic phospholipid membranes is known to correlate with cellular toxicity. In the present study, we measured membrane interaction of the prefibrillar and native states for three variants: the Y31 isoform studied previously, the wild-type protein and the G4R mutant; the latter is observed in progressive myoclonus epilepsy of type 1. In addition to using critical pressure and surface plasmon resonance, we assessed membrane permeabilization by calcein release and electrophysiological measurements. It was demonstrated for the first time that wild-type stefin B and the Y31 isoform are able to form pores in planar lipid bilayers, whereas G4R destroys the bilayer by a non pore-forming process. Similarities to other amyloidogenic proteins and the possible physiological implications of our findings are discussed. PMID- 18397317 TI - Catalytic activation of human glucokinase by substrate binding: residue contacts involved in the binding of D-glucose to the super-open form and conformational transitions. AB - alpha-D-Glucose activates glucokinase (EC 2.7.1.1) on its binding to the active site by inducing a global hysteretic conformational change. Using intrinsic tryptophan fluorescence as a probe on the alpha-D-glucose induced conformational changes in the pancreatic isoform 1 of human glucokinase, key residues involved in the process were identified by site-directed mutagenesis. Single-site W-->F mutations enabled the assignment of the fluorescence enhancement (DeltaF/F(0)) mainly to W99 and W167 in flexible loop structures, but the biphasic time course of DeltaF/F(0) is variably influenced by all tryptophan residues. The human glucokinase-alpha-D-glucose association (K(d) = 4.8 +/- 0.1 mm at 25 degrees C) is driven by a favourable entropy change (DeltaS = 150 +/- 10 J.mol(-1).K(-1)). Although X-ray crystallographic studies have revealed the alpha-d-glucose binding residues in the closed state, the contact residues that make essential contributions to its binding to the super-open conformation remain unidentified. In the present study, we combined functional mutagenesis with structural dynamic analyses to identify residue contacts involved in the initial binding of alpha-d glucose and conformational transitions. The mutations N204A, D205A or E256A/K in the L-domain resulted in enzyme forms that did not bind alpha-D-glucose at 200 mm and were essentially catalytically inactive. Our data support a molecular dynamic model in which a concerted binding of alpha-D-glucose to N204, N231 and E256 in the super-open conformation induces local torsional stresses at N204/D205 propagating towards a closed conformation, involving structural changes in the highly flexible interdomain connecting region II (R192-N204), helix 5 (V181 R191), helix 6 (D205-Y215) and the C-terminal helix 17 (R447-K460). PMID- 18397318 TI - Study of uptake of cell penetrating peptides and their cargoes in permeabilized wheat immature embryos. AB - The uptake of five fluorescein labeled cell-penetrating peptides (Tat, Tat(2), mutated-Tat, peptide vascular endothelial-cadherin and transportan) was studied in wheat immature embryos. Interestingly, permeabilization treatment of the embryos with toluene/ethanol (1 : 20, v/v with permeabilization buffer) resulted in a remarkably higher uptake of cell-penetrating peptides, whereas nonpermeabilized embryos failed to show significant cell-penetrating peptide uptake, as observed under fluorescence microscope and by fluorimetric analysis. Among the cell-penetrating peptides investigated, Tat monomer (Tat) showed highest fluorescence uptake (4.2-fold greater) in permeabilized embryos than the nonpermeabilized embryos. On the other hand, mutated-Tat serving as negative control did not show comparable fluorescence levels even in permeabilized embryos. A glucuronidase histochemical assay revealed that Tat peptides can efficiently deliver functionally active beta-glucuronidase (GUS) enzyme in permeabilized immature embryos. Tat(2)-mediated GUS enzyme delivery showed the highest number of embryos with GUS uptake (92.2%) upon permeabilization treatment with toluene/ethanol (1 : 40, v/v with permeabilization buffer) whereas only 51.8% of nonpermeabilized embryos showed Tat(2)-mediated GUS uptake. Low temperature, endocytosis and macropinocytosis inhibitors reduced delivery of the Tat(2)-GUS enzyme cargo complex. The results suggest that more than one mechanism of cell entry is involved simultaneously in cell-penetrating peptide-cargo uptake in wheat immature embryos. We also studied Tat(2)-plasmid DNA (carrying Act-1GUS) complex formation by gel retardation assay, DNaseI protection assay and confocal laser microscopy. Permeabilized embryos transfected with Tat(2)-plasmid DNA complex showed 3.3-fold higher transient GUS gene expression than the nonpermeabilized embryos. Furthermore, addition of cationic transfecting agent Lipofectamine 2000 to the Tat(2)-plasmid DNA complex resulted in 1.5-fold higher transient GUS gene expression in the embryos. This is the first report demonstrating translocation of various cell-penetrating peptides and their potential to deliver macromolecules in wheat immature embryos in the presence of a cell membrane permeabilizing agent. PMID- 18397319 TI - AKAP12 in astrocytes induces barrier functions in human endothelial cells through protein kinase Czeta. AB - Interactions between astrocytes and blood vessels are essential for the formation and maintenance of the blood-neural barrier (BNB). Astrocyte-derived A-kinase anchor protein 12 (AKAP12) influences BNB formation, but the mechanism of regulation of BNB functions by AKAP12 is not fully understood. We have defined a new pathway of barriergenesis in human retina microvascular endothelial cells (HRMECs) involving astrocytic AKAP12. Treatment of HRMECs with conditioned media from AKAP12-overexpressing astrocytes reduced phosphorylation of protein kinase Czeta (PKCzeta), decreased the levels of vascular endothelial growth factor (VEGF) mRNA and protein, and increased thrombospondin-1 (TSP-1) levels, which led to antiangiogenesis and barriergenesis. Transfection of a small interference RNA targeting PKCzeta decreased VEGF levels and increased TSP-1 levels in HRMECs. Rho is a putative downstream signal of PKCzeta, and inhibition of Rho kinase with a specific inhibitor, Y27632, decreased VEGF levels and increased TSP-1 levels. We therefore suggest that AKAP12 in astrocytes differentially regulates the expression of VEGF and TSP-1 via the inhibition of PKCzeta phosphorylation and Rho kinase activity in HRMECs. PMID- 18397320 TI - Investigation of the interaction between the atypical agonist c[YpwFG] and MOR. AB - Endogenous and exogenous opiates are currently considered the drugs of choice for treating different kinds of pain. However, their prolonged use produces several adverse symptoms, and in addition, many forms of pain are resistant to any kind of therapy. Therefore, the discovery of compounds active towards mu-opioid receptors (MORs) by alternative pharmacological mechanisms could be of value for developing novel classes of analgesics. There is evidence that some unusual molecules can bind opioid receptors, albeit lacking some of the typical opioid pharmacophoric features. In particular, the recent discovery of a few compounds that showed agonist behavior even in the absence of the primary pharmacophore, namely a protonable amine, led to a rediscussion of the importance of ionic interactions in stabilizing the ligand-receptor complex and in activating signal transduction. Very recently, we synthesized a library of cyclic analogs of the endogenous, MOR-selective agonist endomorphin-1 (YPWF-NH(2)), containing a Gly5 bridge between Tyr1 and Phe4. The cyclopeptide c[YpwFG] showed good affinity and agonist behavior. This atypical MOR agonist does not have the protonable Tyr amine. In order to gain more information about plausible mechanisms of interaction between c[YpwFG] and the opioid receptor, we synthesized a selected set of derivatives containing different bridges between Tyr1 and Phe4, and tested their affinities towards mu-opioid receptors. We performed conformational analysis of the cyclopeptides by NMR spectroscopy and molecular dynamics, and investigated plausible, unprecedented modes of interaction with the MOR by molecular docking. The successive quantum mechanics/molecular mechanics investigation of the complexes obtained by the molecular docking procedure furnished a more detailed description of the binding mode and the electronic properties of the ligands. The comparison with the binding mode of the potent agonist JOM-6 seems to indicate that the cyclic endomorphin-1 analogs interact with the receptor by way of an alternative mechanism, still maintaining the ability to activate the receptor. PMID- 18397321 TI - Spectroscopic characterization of the oxyferrous complex of prostacyclin synthase in solution and in trapped sol-gel matrix. AB - Prostacyclin synthase (PGIS) is a member of the cytochrome P450 family in which the oxyferrous complexes are generally labile in the absence of substrate. At 4 degrees C, the on-rate constants and off-rate constants of oxygen binding to PGIS in solution are 5.9 x 10(5) m(-1).s(-1) and 29 s(-1), respectively. The oxyferrous complex decays to a ferric form at a rate of 12 s(-1). We report, for the first time, a stable oxyferrous complex of PGIS in a transparent sol-gel monolith. The encapsulated ferric PGIS retained the same spectroscopic features as in solution. The binding capabilities of the encapsulated PGIS were demonstrated by spectral changes upon the addition of O-based, N-based and C based ligands. The peroxidase activity of PGIS in sol-gel was three orders of magnitude slower than that in solution owing to the restricted diffusion of the substrate in sol-gel. The oxyferrous complex in sol-gel was observable for 24 h at room temperature and displayed a much red-shifted Soret peak. Stabilization of the ferrous-carbon monoxide complex in sol-gel was observed as an enrichment of the 450-nm species over the 420-nm species. This result suggests that the sol-gel method may be applied to other P450s to generate a stable intermediate in the di oxygen activation. PMID- 18397322 TI - Activity of soluble OX40 ligand is enhanced by oligomerization and cell surface immobilization. AB - OX40 ligand (OX40L) and OX40 are typical members of the tumor necrosis factor ligand family and the tumor necrosis factor receptor superfamily, respectively, and are involved in the costimulation and differentiation of T cells. Like other tumor necrosis factor ligands, OX40L is a type II transmembrane protein. Recombinant soluble human OX40L assembles into trimers and is practically inactive despite binding to OX40. However, oligomerization of soluble OX40L trimers by cross-linking with antibodies or by expression as a hexameric fusion protein strongly increased the activity of the ligand. Moreover, a fusion protein of OX40L with a single chain fragment recognizing the tumor stroma antigen fibroblast activation protein showed a cell surface antigen-dependent increase in the activity of the ligand domain of the molecule and thus mimicked the activity of membrane OX40L upon antigen binding. Trimeric single chain OX40L fusion proteins therefore represent a novel type of OX40L-derived immunostimulatory molecule with potentially reduced systemic side effects. PMID- 18397323 TI - The crystal structure of human WD40 repeat-containing peptidylprolyl isomerase (PPWD1). AB - Cyclophilins comprise one of the three classes of peptidylprolyl isomerases found in all eukaryotic and prokaryotic organisms, as well as viruses. Many of the 17 annotated human cyclophilins contain the catalytic domain in tandem with other domains, and many of the specific functions of a particular cyclophilin or its associated domains remain unknown. The structure of the isomerase domain from a spliceosome-associated cyclophilin, PPWD1 (peptidylprolyl isomerase containing WD40 repeat), has been solved to 1.65 A. In the crystal, the N-terminus of one isomerase domain is bound in the active site of a neighboring isomerase molecule in a manner analogous to substrate. NMR solution studies show that this sequence binds to the active site of the cyclophilin, but cannot be turned over by the enzyme. A pseudo-substrate immediately N-terminal to the cyclophilin domain in PPWD1 could have wider implications for the function of this cyclophilin in the spliceosome, where it is located in human cells. PMID- 18397324 TI - A hydrophilic cation-binding protein of Arabidopsis thaliana, AtPCaP1, is localized to plasma membrane via N-myristoylation and interacts with calmodulin and the phosphatidylinositol phosphates PtdIns(3,4,5)P(3) and PtdIns(3,5)P(2). AB - A hydrophilic cation-binding protein, PCaP1, was found to be stably bound to the plasma membrane in Arabidopsis thaliana. PCaP1 was quantified to account for 0.03 0.08% of the crude membrane fractions from roots and shoots. Its homologous protein was detected in several plant species. We investigated the mechanism of membrane association of PCaP1 by transient expression of fusion protein with green fluorescent protein. The amino-terminal sequence of 27 residues of PCaP1 had a potential to localize the fusion protein with green fluorescent protein to the plasma membrane, and the substitution of Gly at position 2 with Ala resulted in the cytoplasmic localization of PCaP1. When PCaP1 was expressed in the in vitro transcription/translation system with [(3)H]myristic acid, the label was incorporated into PCaP1, but not into a mutant PCaP1 with Gly2 replaced by Ala. These results indicate that PCaP1 tightly binds to the plasma membrane via N myristoylation at Gly2. We examined the binding capacity with phosphatidylinositol phosphates (PtdInsPs), and found that PCaP1 selectively interacts with phosphatidylinositol 3,5-bisphosphate and phosphatidylinositol 3,4,5-triphosphate. Competition assay with the N-terminal peptide and mutational analysis revealed that PCaP1 interacts with these two PtdInsPs at the N-terminal part. Interaction of PCaP1 with the membrane and PtdInsPs was not altered in the presence of Ca(2+) at physiological concentrations. Furthermore, calmodulin associated with PCaP1 in a Ca(2+)-dependent manner, and its association weakened the interaction of PCaP1 with PtdInsPs. These results indicate that the N terminal part is essential for both N-myristoylation and interaction with PtdInsPs, and that PCaP1 may be involved in intracellular signalling through interaction with PtdInsPs and calmodulin. PMID- 18397325 TI - Structure-activity relationships of wheat flavone O-methyltransferase: a homodimer of convenience. AB - Wheat flavone O-methyltransferase catalyzes three sequential methylations of the flavone tricetin. Like other flavonoid O-methyltransferases, the protein is a homodimer. We demonstrate, using analytical ultracentrifugation, that perchlorate dissociates the dimer into monomers. The resulting monomers retain all their catalytic capacity, including the ability to catalyze the three successive methylations. We show, using isothermal titration calorimetry, that the binding constant for S-adenosyl-L-methionine does not change significantly as the protein dissociates. The second substrate, tricetin, binds to the dimers but could not be tested with the monomers. CD, UV and fluorescence spectroscopy show that there are substantial changes in the structure of the protein as it dissociates. The fact that there are differences between the monomers and dimers even as the monomers maintain activity may be the result of the very low catalytic capacity of this enzyme. Maximal turnover numbers for the dimers and monomers are only about 6-7 per minute. Even though the binding pockets for S-adenosyl-L methionine, tricetin, selgin and tricin are intact, selection of a catalytically competent structure may be a very slow step during catalysis. PMID- 18397326 TI - Biochemical characteristics of C-terminal region of recombinant chitinase from Bacillus licheniformis: implication of necessity for enzyme properties. AB - The functional and structural significance of the C-terminal region of Bacillus licheniformis chitinase was explored using C-terminal truncation mutagenesis. Comparative studies between full-length and truncated mutant molecules included initial rate kinetics, fluorescence and CD spectrometric properties, substrate binding and hydrolysis abilities, thermostability, and thermodenaturation kinetics. Kinetic analyses revealed that the overall catalytic efficiency, k(cat)/K(m), was slightly increased for the truncated enzymes toward the soluble 4-methylumbelliferyl-N-N'-diacetyl chitobiose or 4-methylumbelliferyl-N-N''-N''' triacetyl chitotriose or insoluble alpha-chitin substrate. By contrast, changes to substrate affinity, K(m), and turnover rate, k(cat), varied considerably for both types of chitin substrates between the full-length and truncated enzymes. Both truncated enzymes exhibited significantly higher thermostabilities than the full-length enzyme. The truncated mutants retained similar substrate-binding specificities and abilities against the insoluble substrate but only had approximately 75% of the hydrolyzing efficiency of the full-length chitinase molecule. Fluorescence spectroscopy indicated that both C-terminal deletion mutants retained an active folding conformation similar to the full-length enzyme. However, a CD melting unfolding study was able to distinguish between the full-length and truncated mutant molecules by the two phases of apparent transition temperatures in the mutants. These results indicate that up to 145 amino acid residues, including the putative C-terminal chitin-binding region and the fibronectin (III) motif of B. licheniformis chitinase, could be removed without causing a seriously aberrant change in structure and a dramatic decrease in insoluble chitin hydrolysis. The results of the present study provide evidence demonstrating that the binding and hydrolyzing of insoluble chitin substrate for B. licheniformis chitinase was not dependent solely on the putative C-terminal chitin-binding region and the fibronectin (III) motif. PMID- 18397327 TI - Factors influencing RNA degradation by Thermus thermophilus polynucleotide phosphorylase. AB - At the optimal temperature (65 degrees C), Thermus thermophilus polynucleotide phosphorylase (Tth PNPase), produced in Escherichia coli cells and isolated to functional homogeneity, completely destroys RNAs that possess even a very stable intramolecular secondary structure, but leaves intact RNAs whose 3' end is protected by chemical modification or by hybridization with a complementary oligonucleotide. This allows individual RNAs to be isolated from heterogeneous populations by degrading unprotected species. If oligonucleotide is hybridized to an internal RNA segment, the Tth PNPase stalls eight nucleotides downstream of that segment. This allows any arbitrary 5'-terminal fragment of RNA to be prepared with a precision similar to that of run-off transcription, but without the need for a restriction site. In contrast to the high Mg(2+) requirements of mesophilic PNPases, Tth PNPase retains significant activity when the free Mg(2+) concentration is in the micromolar range. This allows minimization of the Mg(2+) catalysed nonenzymatic hydrolysis of RNA when phosphorolysis is performed at a high temperature. This capability of Tth PNPase for fully controlled RNA phosphorolysis could be utilized in a variety of research and practical applications. PMID- 18397328 TI - Erythromycin-heteroresistant methicillin-resistant Staphylococcus aureus isolates from Turkey. PMID- 18397329 TI - Molecular analysis of drug-resistant Mycobacterium tuberculosis isolates collected in central Poland. AB - In total, 48 Mycobacterium tuberculosis clinical isolates from 48 patients with drug-resistant pulmonary tuberculosis (TB) in central Poland during 2004 were analysed by spoligotyping and IS6110-Mtb1/Mtb2 PCR. Twelve (25%) isolates were clustered by spoligotyping in combination with IS6110-Mtb1/Mtb2 PCR. Patients from whom these isolates were obtained were assumed to have developed TB as a result of recent transmission. Spoligotyping, used alone, overestimated the number of clustered isolates. However, combined use of spoligotyping and IS6110 Mtb1/Mtb2 PCR was an efficient approach for revealing clonal relatedness among M. tuberculosis isolates. PMID- 18397330 TI - A comparison of metronidazole and single-dose ornidazole for the treatment of dientamoebiasis. AB - Recent reports of the pathogenic potential of Dientamoeba fragilis have underlined the need for an effective treatment against this colon-dwelling protozoan. Metronidazole is a well-known and commonly used anti-protozoal agent, but another 5-nitroimidazole derivative, ornidazole, may be preferable, where available, because of its longer half-life and fewer side-effects. This study compared the efficacies of metronidazole and ornidazole in a group of 112 patients with dientamoebiasis. Patients were randomised into two treatment groups: group 1 (n = 56) received metronidazole for 5 days, 20 mg/kg/day for children and 1.5 g/day for adults, in three oral doses, while group 2 (n = 56) received a single oral dose of ornidazole, 30 mg/kg for children and 2 g for adults. Stool samples were examined on the seventh and 14th days after treatment, and clinical symptoms were recorded to evaluate the efficacy of treatment. A statistically significant difference was recorded between the efficacies of ornidazole and metronidazole, both parasitologically (92.9% vs. 69.6%, p 0.001) and clinically (96.4% vs. 76.8%, p 0.001). Patients in the metronidazole group reported more side-effects than patients in the ornidazole group, none of whom required termination of treatment. These results suggest that single-dose ornidazole may be an important alternative agent for the treatment of dientamoebiasis. PMID- 18397331 TI - Harmonised monitoring of antimicrobial resistance in Salmonella and Campylobacter isolates from food animals in the European Union. AB - Many Member States of the European Union (EU) currently monitor antimicrobial resistance in zoonotic agents, including Salmonella and Campylobacter. According to Directive 2003/99/EC, Member States shall ensure that the monitoring provides comparable data on the occurrence of antimicrobial resistance. The European Commission asked the European Food Safety Authority to prepare detailed specifications for harmonised schemes for monitoring antimicrobial resistance. The objective of these specifications is to lay down provisions for a monitoring and reporting scheme for Salmonella in fowl (Gallus gallus), turkeys and pigs, and for Campylobacter jejuni and Campylobacter coli in broiler chickens. The current specifications are considered to be a first step towards a gradual implementation of comprehensive antimicrobial resistance monitoring at the EU level. These specifications propose to test a common set of antimicrobial agents against available cut-off values and a specified concentration range to determine the susceptibility of Salmonella and Campylobacter. Using isolates collected through programmes in which the sampling frame covers all epidemiological units of the national production, the target number of Salmonella isolates to be included in the antimicrobial resistance monitoring per Member State per year is 170 for each study population (i.e., laying hens, broilers, turkeys and slaughter pigs). The target number of Campylobacter isolates to be included in the antimicrobial resistance monitoring per Member State per year is 170 for each study population (i.e., broilers). The results of the antimicrobial resistance monitoring are assessed and reported in the yearly national report on trends and sources of zoonoses, zoonotic agents and antimicrobial resistance. PMID- 18397332 TI - Detection and molecular characterisation of plasmidic AmpC beta-lactamases in Klebsiella pneumoniae isolates from a tertiary-care hospital in Dublin, Ireland. AB - This study determined the types of AmpC enzymes produced by Klebsiella pneumoniae isolates resistant to third-generation cephalosporins and the clonality of these isolates. The presence of AmpC enzymes was identified by cephalosporin cloxacillin synergy tests. Genes encoding AmpC enzymes were characterised by PCR and sequencing. Pulsed-field gel electrophoresis (PFGE) was used to type the isolates. Fifteen K. pneumoniae isolates were positive for bla(AmpC), 13 were positive for bla(ACC-1) and two were positive for bla(DHA-1). Production of the DHA-1 enzyme was inducible. The ampR gene was identified upstream of the bla(DHA 1) gene. PFGE demonstrated the polyclonal origin of the isolates carrying bla(ACC 1). PMID- 18397333 TI - Comparison of phenotypic methods for penicillinase detection in Staphylococcus aureus. AB - Penicillinase testing is required for Staphylococcus aureus isolates with a penicillin MIC of /=2.5 mg/kg prolonged survival significantly. In addition, the two doses evaluated for reductions in fungal burden (5 and 15 mg/kg) revealed fungicidal activity. As prophylaxis, both aminocandin and caspofungin 5 and 30 mg/kg prolonged survival when given 7 days before inoculation. Aminocandin and caspofungin 30 mg/kg were both able to prolong survival when the interval between dose and inoculation was increased to 10 days. When this interval was extended to 14 days, only aminocandin 30 mg/kg prolonged survival and reduced fungal burden. These results demonstrate that single doses of aminocandin are effective as treatment and prophylaxis, and suggest that extended interval dosing may be a useful strategy for treating invasive candidiasis. PMID- 18397336 TI - A comparative in-vitro evaluation of resistance selection after exposure to teicoplanin, vancomycin, linezolid and quinupristin-dalfopristin in Staphylococcus aureus and Enterococcus spp. AB - The ability of breakpoint and serum concentrations of teicoplanin, vancomycin, linezolid and quinupristin-dalfopristin to select resistance was compared for isolates of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin resistant S. aureus (MRSA), Enterococcus faecalis and Enterococcus faecium. Mutation frequencies were always <10(-10), except for two isolates grown in the presence of teicoplanin at the trough serum concentration. After multistep selection, linezolid selected for resistance in staphylococci and enterococci, and serial exposure to certain concentrations of linezolid was more likely to select for stable resistance in MRSA, MSSA and enterococci than was exposure to glycopeptides and quinupristin-dalfopristin. PMID- 18397337 TI - Cell proliferation, apoptosis and mitochondrial damage in rat B50 neuronal cells after cisplatin treatment. AB - OBJECTIVES: Cisplatin (cisPt) is used as a chemotherapeutic agent for the treatment of a variety of human tumours; more recently, it has been demonstrated that tumour cell exposure to cisPt ultimately results in apoptosis, but the mechanism by which nuclear cisPt/DNA generates the cytoplasmic cascade of events involved has not been clarified. We have investigated the effects of cisPt on proliferation in the neuronal cell line B50, with particular attention being given to understand whether mitochondria are a target of cisPt and their involvement in the apoptotic process. MATERIALS AND METHODS: Rat neuronal B50 cells were used to investigate the mechanisms of cisPt-induced cytotoxicity; this line has been used as a model system for neurotoxicity in vivo. RESULTS: Changes in proliferation, induction of apoptosis, activation of caspase-3 and DNA fragmentation were observed in the cells, as well as morphological and biochemical alterations of mithocondria. Activation of caspase-9 confirmed that mitochondria are a target of cisPt. CONCLUSION: CisPt exerts cytotoxic effects in the neuronal B50 cell line via a caspase-dependent pathway with mitochondria being central relay stations. PMID- 18397338 TI - Multiple imputation methods for treatment noncompliance and nonresponse in randomized clinical trials. AB - Randomized clinical trials are a powerful tool for investigating causal treatment effects, but in human trials there are oftentimes problems of noncompliance which standard analyses, such as the intention-to-treat or as-treated analysis, either ignore or incorporate in such a way that the resulting estimand is no longer a causal effect. One alternative to these analyses is the complier average causal effect (CACE) which estimates the average causal treatment effect among a subpopulation that would comply under any treatment assigned. We focus on the setting of a randomized clinical trial with crossover treatment noncompliance (e.g., control subjects could receive the intervention and intervention subjects could receive the control) and outcome nonresponse. In this article, we develop estimators for the CACE using multiple imputation methods, which have been successfully applied to a wide variety of missing data problems, but have not yet been applied to the potential outcomes setting of causal inference. Using simulated data we investigate the finite sample properties of these estimators as well as of competing procedures in a simple setting. Finally we illustrate our methods using a real randomized encouragement design study on the effectiveness of the influenza vaccine. PMID- 18397339 TI - Genome-wide analysis of subependymomas shows underlying chromosomal copy number changes involving chromosomes 6, 7, 8 and 14 in a proportion of cases. AB - Subependymomas (SE) are slow-growing brain tumors that tend to occur within the ventricles of middle-aged and elderly adults. The World Health Organization classifies these tumors within the ependymoma group. Previous limited analysis of this tumor type had not revealed significant underlying cytogenetic abnormalities. We have used microarray comparative genomic hybridization to study a series of SE (n = 12). A whole-genome array at 0.97-Mb resolution showed copy number abnormalities in five of 12 cases (42%). Two cases (17%) showed regions of loss on chromosome 6. More detailed analysis of all cases using a chromosome 6 tile-path array confirmed the presence of overlapping regions of loss in only these two cases. One of these cases also showed trisomy chromosome 7. Monosomy of chromosome 8 was seen in a further two cases (17%), and a partial loss on chromosome 14 was observed in one additional case. This is the first array-based, genome-wide study of SE. The observation that five of 12 cases examined (42%) at 0.97-Mb resolution showed chromosomal copy number abnormalities is a novel finding in this tumor type. PMID- 18397340 TI - Relationship between ADAMTS13 activity in clinical remission and the risk of TTP relapse. AB - Idiopathic thrombotic thrombocytopenic purpura (TTP) is characterized by frequent recurrences. Effective screening for relapses will enable intervention prior to overt episodes of TTP. The present study used a modified assay to detect ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type 1 motif, 13) activity as low as 0.5%. This analytical improvement permits adequate measurement of ADAMTS13 activity levels in 97% of remission samples used for statistical modelling. ADAMTS13 activity and ADAMTS13 antibody (IgG) were measured in 157 serial samples prospectively collected from 24 TTP patients during periods of clinical remission. These patients were followed-up quarterly for an average of 23 months, during which time nine episodes of TTP relapse occurred among six patients. Finally, logistic regression modelling was used to define the relationship between ADAMTS13 activity levels (0.5-100%) and the probability of TTP relapses. Our data demonstrated that lower ADAMTS13 activity and younger age were significantly associated with higher risk of relapse in the 3 months after specimens were taken. In contrast, ADAMTS13 antibody IgG levels were not predictive of TTP relapses. Identification of low ADAMTS13 activity during clinical remission as a key risk factor for TTP relapses provides a new screening strategy to identify patients who may benefit from prophylactic therapy prior to disease relapses. PMID- 18397341 TI - High density genome-wide DNA profiling reveals a remarkably stable profile in hairy cell leukaemia. AB - Hairy cell leukaemia (HCL) is a rare B-cell neoplasm for which the molecular mechanisms are largely unknown. High-density genome-wide DNA profiling was performed with Affymetrix 250K arrays to analyse copy number (CN) changes and loss of heterozygosity (LOH) in 16 cases of HCL. Four of 16 cases (25%) demonstrated gross non-recurrent CN deletions. Within the affected regions, we identified genes involved in bone marrow fibrosis (FGF12) and response to treatment (TP53) in individual cases. Large regions (> 5 Mb) of LOH without any concomitant DNA CN changes were identified in 5/16 (31%) HCL and were indicative of uniparental disomy UD. The germline origin of UD was demonstrated in one case for which a matched normal sample was available. Overall analysis of LOH showed that identical loci were recurrently targeted in chromosomes 1, 2 and 6. As a whole, however, HCL showed a remarkably stable genome. This finding adds to several other features that are unique to HCL among mature B-cell tumours. PMID- 18397342 TI - Marrow osteopontin level as a prognostic factor in acute myeloid leukaemia. PMID- 18397343 TI - Functional analysis of JAK3 mutations in transient myeloproliferative disorder and acute megakaryoblastic leukaemia accompanying Down syndrome. AB - JAK3 mutations have been reported in transient myeloproliferative disorder (TMD) as well as in acute megakaryoblastic leukaemia of Down syndrome (DS-AMKL). However, functional consequences of the JAK3 mutations in TMD patients remain undetermined. To further understand how JAK3 mutations are involved in the development and/or progression of leukaemia in Down syndrome, additional TMD patients and the DS-AMKL cell line MGS were screened for JAK3 mutations, and we examined whether each JAK3 mutation is an activating mutation. JAK3 mutations were not detected in 10 TMD samples that had not previously been studied. Together with our previous report we detected JAK3 mutations in one in 11 TMD patients. Furthermore, this study showed for the first time that a TMD patient derived JAK3 mutation (JAK3(I87T)), as well as two novel JAK3 mutations (JAK3(Q501H) and JAK3(R657Q)) identified in an MGS cell line, were activating mutations. Treatment of MGS cells and Ba/F3 cells expressing the JAK3 mutants with JAK3 inhibitors significantly decreased their growth and viability. These results suggest that the JAK3 activating mutation is an early event during leukaemogenesis in Down syndrome, and they provide proof-of-principle evidence that JAK3 inhibitors would have therapeutic effects on TMD and DS-AMKL patients carrying activating JAK3 mutations. PMID- 18397344 TI - Early stage chronic lymphocytic leukaemia carrying unmutated IGHV genes is at risk of recurrent infections during watch and wait. PMID- 18397346 TI - Keloid scars--an avoidable complication of vaccination. PMID- 18397345 TI - p38 mitogen-activated protein kinase inhibitor LY2228820 enhances bortezomib induced cytotoxicity and inhibits osteoclastogenesis in multiple myeloma; therapeutic implications. AB - The interaction between multiple myeloma (MM) cells and the bone marrow (BM) microenvironment induces proliferation and survival of MM cells, as well as osteoclastogenesis. This study investigated the therapeutic potential of novel p38 mitogen-activated protein kinase (p38MAPK) inhibitor LY2228820 (LY) in MM. Although cytotoxicity against MM cell lines was modest, LY significantly enhanced the toxicity of bortezomib by down-regulating bortezomib-induced heat shock protein 27 phosphorylation. LY inhibited interleukin-6 secretion from long term cultured-BM stromal cells and BM mononuclear cells (BMMNCs) derived from MM patients in remission. LY also inhibited macrophage inflammatory protein-1alpha secretion from patient MM cells and BMMNCs as well as normal CD14 positive osteoclast precursor cells. Moreover, LY significantly inhibited in vitro osteoclastogenesis from CD14 positive cells induced by macrophage-colony stimulating factor and soluble receptor activator of nuclear factor-kappaB ligand. Finally, LY also inhibited in vivo osteoclatogenesis in a severe combined immunodeficiency mouse model of human MM. These results suggest that LY represents a promising novel targeted approach to improve MM patient outcome both by enhancing the effect of bortezomib and by reducing osteoskeletal events. PMID- 18397347 TI - Systemic air embolism after cardiopulmonary resuscitation in a preterm infant. AB - We report a preterm infant with extensive systemic air embolism after cardiopulmonary resuscitation for cardiac arrest due to an occluding thrombus in the inferior vena cava. After excluding other potential causes (air infusion, necrotizing enterocolitis or pulmonary leakage syndrome), we postulate that the pressure gradient needed for air embolism to occur is related to the resuscitation procedure. An important clue of air embolism was noted on the chest X-ray taken before death showing intracardial air. CONCLUSION: Systemic air embolism may occur as a very rare complication after cardiopulmonary resuscitation. PMID- 18397348 TI - Predictive value of Apgar score in infants with very low birth weight. AB - BACKGROUND: Accurate prediction of the prognosis of infants with very low birth weight is beneficial both for their parents and for healthcare professionals. METHODS: This was a population-based study of all low-birth-weight infants admitted to neonatal units in one region in Japan. The Apgar score at 1 and 5 min and neonatal mortality were retrospectively analyzed to obtain the predictive values of the scores. The results were stratified into two categories by birth weight and three time periods (1980-1986, 1987-1993 and 1994-2000). RESULTS: The predictive values improved in the later years, and therefore only the data obtained in the period 1994-2000 were used. A score of less than 5 at 5 min appears to be a good predictor of neonatal mortality in infants with a birth weight between 1500 g and 2499 g (positive likelihood ratio, 17.59 [95% confidence interval (CI) 12.68-24.40]); however, there is no evidence that the score is a good predictor of neonatal mortality in infants with very low birth weight. CONCLUSION: In infants with a low birth weight between 1500 g and 2499 g, an Apgar score at 5 min of less than 5 is a good predictor of neonatal mortality. The score is not useful in predicting the short-term prognosis of very low-birth weight infants. PMID- 18397349 TI - Adiponectin: an intriguing hormone for paediatricians. AB - Adiponectin, a protein hormone produced by adipocytes, is also found in breast milk, which in turn is implicated in childhood obesity prevention. Although a biological role for adiponectin has not been firmly established, clinical and experimental research indicates that it regulates lipid and glucose metabolism, affects foetal development, and exerts anti-inflammatory and antiatherogenic effects. CONCLUSION: This review demonstrates an emerging interest of paediatric research on adiponectin. A better understanding of adiponectin's bioactivity might clarify whether breast milk indeed prevents childhood obesity. PMID- 18397351 TI - Shaken baby syndrome and a baby's cry. AB - The aim of this study was to investigate the relationship between crying of an infant and inflicted head injury by shaking and/or impact. During the period between January 1, 1997 and December 31, 2003, 26 cases of shaken baby syndrome (SBS) were identified in Estonia. The incidence of SBS was 28.7 per 100,000 children under 1 year of age during the whole study period. In this group there were four children from twin pairs: two twin boys and a girl from a twin pair and a boy from another twin pair. This represents 15.4% of the 26 cases. Twins in Estonia represent 2.12% of infant births. The mean age on admission was 3.9 months. According to outpatient records almost all parents (88.5%) in the study group (23/26) had contacted their family physicians and other specialists because of excessive crying or irritability of the baby prior to the admission to the hospital with SBS or death. We found that the time curve of crying was similar to the curve of highest incidence of cases of SBS except the crying curve began earlier. CONCLUSION. Our data confirm that the families with twins are at additional risk for SBS and parent's complaints of excessive crying of their infants should be taken as signal that parents need to be carefully counselled. PMID- 18397350 TI - Neonatal hyperbilirubinemia increases intestinal protein permeability and the prevalence of cow's milk protein intolerance. AB - AIMS: Bilirubin is a newly discovered modulator of the gut barrier in vitro and in vivo. We studied the effect of bilirubin on the serosal to mucosal intestinal permeability in vivo. We also investigated the prevalence of cow's milk protein intolerance (CMPI) in infants with moderate hyperbilirubinemia versus matched controls. METHODS: Faecal alpha 1 antitrypsin (a1AT) was used to monitor intestinal protein loss; a large cohort was prospectively followed for 12 months for sign and symptoms of CMPI. RESULTS: Neonates with hyperbilirubinemia had higher stool excretion of a1AT than controls (0.68 +/- 0.28 mg/g vs. 0.25 +/- 0.11 mg/g; p < 0.01). Faecal a1AT correlates with total serum bilirubin (TSB) (r = 0.85; p < 0.01). Also, in the first 12 months of life, formerly hyperbilirubinemic infants had an higher prevalence of CMPI (14/353 vs. 4/339; chi2= 4.018, p = 0.045). CONCLUSIONS: Neonatal hyperbilirubinemia increases stool protein loss and is also a mild risk factor for CMPI. PMID- 18397352 TI - Nomogram of the cerebellar vermis height at birth in small-for-gestational-age neonates. AB - AIM: To establish a nomogram of the height of the cerebellar vermis at birth in small-for-gestational-age neonates. METHODS: Eighty-five consecutively born healthy singletons who were small for gestational age were enrolled in a cross sectional observational study. The height of the cerebellar vermis was measured by ultrasonography in the midsagittal plane. Reference intervals for the vermian height at 5th, 50th and 95th centiles were estimated at any week of gestational age using a mean and standard deviation model based on least squares polynomial regression. RESULTS: All attempts at obtaining neonatal vermian height were successful. Mean (standard deviation) maternal age was 29.6 (4.7) years; median parity was 1 (range, 1-3). No difference was found between males and females for the vermian height (mean difference [95% confidence interval], -0.03 [-0.16; 0.10] cm). Mean vermian height (in cm) against gestational age (in weeks) was suitably modelled by a quadratic-cubic polynomial (r2= 0.790, p < 0.0001) as 0.712 + 0.00129 x gestational age2- 0.0000053 x gestational age3 (SD =-1.009 + 0.0504 x gestational age -0.000014 x gestational age3). CONCLUSION: A nomogram of the height of the cerebellar vermis at birth against gestational age was established for use in small-for-gestational-age neonates. PMID- 18397353 TI - Paediatric poisonings treated in one Finnish main university hospital between 2002 and 2006. AB - AIM: Acute poisonings are a major cause of morbidity among children. This study aims to describe the incidence and nature of emergency visits for acute paediatric poisoning among Finnish children. METHODS: All patients younger than 16 years admitted to the Tampere University Hospital's emergency department with a diagnosis of poisoning during 2002-2006 were identified from the Hospital Information System using the International Classification of Diseases (ICD-10). RESULTS: Altogether 369 emergency visits were diagnosed with poisoning, the overall incidence being 8.1 per 10 000 person-years (95% CI 7.3-9.0). A majority of patients were adolescents aged 10-15 years (48%) and children under 5 years (45%). Boys represented 55% of the cases. Nonpharmaceutical agents were suspected to be the cause in 60.4% and pharmaceuticals in 30.6% of the intoxications. Multiple agents were involved in 8.4% of the cases. Ethanol was the agent in 30.9% of the poisonings. Most patients (78.9%) were hospitalized (median length of stay 1 day). Overall mortality was 0.3%. CONCLUSION: Acute paediatric poisonings represent a relatively frequent problem in Finland, and remain a life threatening problem. The high proportion of alcohol poisonings highlights the necessity to develop more effective primary prevention programs. PMID- 18397354 TI - Inhaled corticosteroids do not affect behaviour. AB - AIM: To determine whether children with asthma and on inhaled corticosteroids have more behavioural problems, such as aggressiveness and hyperactivity, as compared with healthy controls and with children under medical care because of other disorders. METHODS: Questionnaires were given to three groups of children: a group of asthmatic children with inhaled corticosteroids (ICS), a group of children attending the ear, nose and throat (ENT) outpatient clinic and the healthy controls. Included were questions about health, medication use, demographical data and about behaviour, including the child behaviour check list (CBCL) and questions about attention deficit hyperactivity disorder (ADHD). RESULTS: Forty asthmatic children on ICS, 50 children visiting the ENT outpatient clinic and 183 healthy controls were studied. The total CBCL and mean ADHD scores of the children on ICS were 28.1 and 9.1, which were both significantly higher than the scores of the healthy controls (20.4 and 7.1), but not when compared with the ENT outpatient group (26.2 and 8.6). Further analysis revealed statistically significant differences between the ICS group and healthy controls in CBCL-axes. There were, however, no differences between the ENT group and the ICS on one side and the healthy controls on the other. CONCLUSION: There is a difference in behaviour between healthy children and asthmatic children on ICS, but not when compared with children visiting the ENT department. Although hyperactivity, aggressiveness and anxiety might occur in children on ICS, this is probably caused by individual susceptibility. Being under specialist care can possibly explain behavioural differences between children on ICS and healthy controls. PMID- 18397355 TI - Neutrophils and red blood cells in the cerebrospinal fluid of newborns. PMID- 18397356 TI - Spinal cord injury at birth: a hidden causative factor. AB - A case of perinatally acquired spinal cord injury (SCI) is presented. The foetus was vigorous until birth, the breech presented and delivery was performed by a non-traumatic Caesarean section. The infant displayed symptoms of severe SCI but diagnosis was delayed due to severe co-morbidity. Diagnostic considerations are briefly reviewed. Ventilatory support was withdrawn at the age of 20 days when the infant had still not exhibited any respiratory effort or spontaneous movements. Autopsy revealed a serious congenital malalignment of the upper cervical vertebrae and at the histological examination extensive reactive changes were observed in the same area. To our knowledge such findings have not been published previously. CONCLUSION: In cases of serious perinatally acquired SCI, claim of malpractice is often apparent. In this case a hidden congenital malformation of the cervical vertebrae was revealed, highlighting the need of careful postmortem examinations in such cases. PMID- 18397357 TI - Establishment of nomograms and reference ranges for intra-cranial ventricular dimensions and ventriculo-hemispheric ratio in newborns by ultrasonography. AB - AIM: To establish normal ranges for the lateral, third, and fourth ventricular dimensions and the ventriculo-hemispheric ratio (VHR) in neonates using cranial ultrasonography. METHOD: Intra-cranial ventricles were assessed ultrasonographically using anterior fontanelle as acoustic window. Data were analyzed to determine correlation, coefficient of determination (R2), regression equations and plotted against gestational age (GA). RESULTS: Of total 1483 neonates (25-42 w GA), 372(25%) had GA < 34 weeks. GA strongly correlated with anterior horn width (AHW; 0.92), thalamo-occipital distance (TOD; 0.94), and third (0.78) and fourth (0.89) ventricular widths. The R2 values were 0.85, 0.88, 0.66 and 0.80, respectively. VHR had weak correlation (0.12, R(2)= 0.005) with GA and stayed constant (0.27-0.28) across GA. Nomograms of AHW, TOD, and third and fourth ventricular widths were constructed with respect to GA. CONCLUSION: Intra cranial ventricles increase in size with GA but this increase is limited to only a few mm and is proportional to increment in brain tissue. Nomograms and regression equations of AHW, TOD and widths of third and fourth ventricles can assist in objectively assessing ventricular sizes. The almost uniform VHR can be used to screen ventricle size while suspecting hydrocephalus. PMID- 18397358 TI - TCF7L2 polymorphisms are associated with type 2 diabetes in Khatri Sikhs from North India: genetic variation affects lipid levels. AB - Recently, the transcription factor-7-like 2 (TCF7L2) gene has been identified as the most important type 2 diabetes mellitus (T2DM) susceptibility gene. Common intronic polymorphisms in this gene have been found to be strongly associated with T2DM susceptibility showing marked reproducibility in multiple populations. The purpose of this study was to confirm the reported association of six TCF7L2 variants in a Khatri Sikh diabetic sample from North India. We genotyped six associated SNPs in a case-control sample consisting of 556 T2DM cases and 537 controls. We also examined the impact of these variants on body mass index (BMI), waist to hip ratio (WHR), fasting insulin, and glucose and lipid levels. We report replication of association of four of the six SNPs with T2DM in this Khatri Sikh sample [rs7903146, (p = 0.010); rs11196205, (p = 0.011); rs10885409, (p = 0.002) and rs4918789, (p = 0.029)], under a dominant model conferring odds ratios (ORs) of 1.39, 1.44, 1.57 and 1.36, respectively. Haplotype analysis provided further evidence of association by showing a significant difference between cases and controls as revealed by the global omnibus test (chi(2)= 19.36; p = 0.0036). Multiple linear regression analysis also revealed the risk allele carriers of three of four significant SNPs (rs7903146, rs11196205, rs10885409) to be significantly associated with increased fasting total cholesterol (p value = 0.019, 0.025, 0.006) and LDL cholesterol levels (p value = 0.021, 0.018, 0.005), respectively. Our findings confirm that the TCF7L2 gene is a major risk factor for development of T2DM in Khatri Sikhs. It also provides new information about the significant impact of TCF7L2 gene variants on plasma cholesterol levels that appear to be independent of BMI. PMID- 18397359 TI - Preference-weighted health-related quality of life measures and substance use disorder severity. AB - AIMS: Examine the validity of preference-weighted health-related quality of life measures in a sample of substance use disorder (SUD) patients. The implications of cost-utility analyses (CUAs) of SUD interventions are discussed. DESIGN: Cross sectional analysis of subjects seeking SUD treatment. SETTING: Seven SUD treatment centers in a medium-sized Midwestern metropolitan area in the United States. PARTICIPANTS: Data from 574 SUD subjects were analyzed from a study to test interventions to improve linkage and engagement with substance abuse treatment. MEASUREMENTS: Subjects completed the following preference-weighted measures: self-administered Quality of Well-Being scale (QWB-SA) and Medical Outcomes Study SF-12 (standard gamble weighted or SF-12 SG); and clinical measures: Addiction Severity Index (ASI) and a symptom checklist based on the DSM IV. FINDINGS: In unadjusted analyses, the QWB-SA was correlated significantly with six of seven ASI subscales and the SF-12 SG was correlated with four of seven. In adjusted analyses, both preference-weighted measures were significantly correlated with diagnostic, physical health, mental health and drug use measures, but not with legal or alcohol use measures. The QWB-SA was also correlated with employment problems and the SF-12 SG was correlated with family/social problems. CONCLUSIONS: This study generally supports the construct validity of preference weighted health-related quality of life measures in SUD patients. However, the QWB-SA and SF-12 SG did not correlate with all ASI scales. Cost-benefit analysis may be preferable when policy-makers are interested in evaluating the full range of SUD intervention outcomes. PMID- 18397360 TI - Does alcohol increase the risk of overdose death: the need for a translational approach. AB - BACKGROUND: We argue for a translational approach to addiction science, using an important current research question as a case study. CASE STUDY: What is the evidence in support of the hypothesis that alcohol increases the risk of a heroin/opiate overdose through a pharmacological interaction? FINDINGS: The positive epidemiological evidence shows that opiate overdose deaths rarely involve a single drug; that alcohol is the most common other drug involved; that there is a negative association between alcohol and morphine concentration at post mortem; and that post-mortem levels of morphine are often below the levels expected of highly tolerant individuals. The evidence is consistent with the hypothesis that heroin users who drink may require less heroin to overdose than those who do not drink (all other factors being equal) because of a pharmacological interaction. However, the evidence is consistent with, and does not rule out, other causal (and non-causal) pathways. Alcohol could be associated negatively with tolerance, or confounded by other factors. Experimental evidence is required which is unlikely to be obtained through further epidemiological study or through randomized clinical trials. CONCLUSIONS: We believe that animal models could provide the key evidence to test the hypothesis for a 'pharmacodynamic' or 'pharmacokinetic' interaction, which could be corroborated in clinical challenge studies and epidemiological studies. Such a translational approach demands greater collaboration between addiction scientists from basic to applied science and from neuroscience to social science, and would be able to address other key research questions and hypotheses in addiction. PMID- 18397361 TI - Arousability in schizophrenia: relationship to emotional and physiological reactivity and symptom severity. AB - OBJECTIVE: Socioenvironmental stressors have been linked with increased symptom severity and relapse in those with schizophrenia. However, little is known about how individual differences in stress reactivity may contribute to these outcomes. METHOD: This study examined the association between the temperament characteristic of arousability and changes in negative affect and cardiovascular activity during a challenge task in 58 in-patients with diagnosis of schizophrenia and 21 controls. RESULTS: In the patient group, levels of arousability were significantly associated with increases in negative affect in response to the task and greater severity of affective symptoms. Levels of arousability were associated with decreased heart rate during the challenge task in our patient group. CONCLUSION: These findings suggest that greater attention be given to individual differences, such as temperament and personality characteristics, and their role in the experience of stressors, including emotional and physiological response, as well as symptom development. PMID- 18397362 TI - Suicidal status during antidepressant treatment in 789 Sardinian patients with major affective disorder. AB - OBJECTIVE: Relationships between antidepressant treatment and suicidality remain uncertain in major depressive disorder (MDD), and rarely evaluated in bipolar disorder (BPD). METHOD: We evaluated changes in suicidality ratings (Hamilton Depression Rating Scale item-3) at the start and after 3.59 +/- 2.57 months of sustained antidepressant treatment in a systematically assessed clinical sample (n = 789) of 605 patients with MDD, 103 patients with BPD-II and 81 patients with BPD-I (based on DSM-IV; 68.1% women; aged 44.3 +/- 16.1 years), comparing suicidal vs. non-suicidal and recovered vs. unrecovered initially suicidal patients. RESULTS: Suicidal patients (103/789, 16.5%; BPD/MDD risk: 2.2) were more depressed and were ill longer. During treatment, 81.5% of suicidal patients became non-suicidal; 0.46% of 656 initially non-suicidal patients reported new suicidal thoughts, with no new attempts. Becoming non-suicidal was associated with greater depression severity and greater improvement. CONCLUSION: Suicidal ideation was prevalent in patients with depressed major affective disorder, but most of the initially suicidal patients became non-suicidal with antidepressant treatment, independent of diagnosis, treatment type or dose. PMID- 18397363 TI - The contribution of TMS to frontotemporal dementia variants. AB - OBJECTIVE: Frontotemporal lobar degeneration (FTLD) includes different heterogeneous conditions mainly characterized by personality changes and cognitive deficits in language and executive functions; movement disorders have also been associated with FTLD. The present study aimed to measure the primary motor cortex (M1) inhibitory and facilitatory functions in patients affected by FTLD. MATERIALS AND METHODS: The study included 17 FTLD patients, 8 age-matched healthy controls and 8 Alzheimer's disease (AD) patients. Transcranial magnetic stimulation (TMS) was used to study intracortical inhibition (ICI) and facilitation (ICF) by using a double-pulse paradigm. RESULTS: FTLD patients were comparable with controls and AD patients for ICI and ICF. Corticobasal degeneration (CBD) patients presented significant reduced inhibition at ISI3; moreover two out of seven CBD patients had only ipsilateral responses. DISCUSSION: The present study reveals a selective impairment of M1 ICI inhibitory response in CBD, which may help in distinguishing among the FTLD clinical spectrum. PMID- 18397364 TI - Docking is not a prerequisite but a temporal constraint for fusion of secretory granules. AB - We examined secretory granule dynamics using total internal reflection fluorescence microscopy in normal pancreatic beta cells and their mutants devoid of Rab27a and/or its effector, granuphilin, which play critical roles in the docking and recruitment of insulin granules to the plasma membrane. In the early phase of glucose stimulation in wild-type cells, we observed marked fusion of granules recruited from a relatively distant area, in parallel with that from granules located underneath the plasma membrane. Furthermore, despite a lack of granules directly attached to the plasma membrane, both spontaneous and evoked fusion was increased in granuphilin-null cells. In addition to these granuphilin null phenotypes, Rab27a/granuphilin doubly deficient cells showed the decreases in granules located next to the docked area and in fusion from granules near the plasma membrane in the early phase of glucose-stimulated secretion, similar to Rab27a-mutated cells. Thus, the two proteins play nonoverlapping roles in insulin exocytosis: granuphilin acts on the granules underneath the plasma membrane, whereas Rab27a acts on those in a more distal area. These findings demonstrate that, in contrast to our conventional understanding, stable attachment of secretory granules to the plasma membrane is not prerequisite but temporally inhibitory for both spontaneous and evoked fusion. PMID- 18397365 TI - Adenosine A1 receptors inhibit GABAergic transmission in rat tuberomammillary nucleus neurons. AB - The adenosinergic modulation of GABAergic spontaneous miniature inhibitory postsynaptic currents (mIPSCs) was investigated in mechanically dissociated rat tuberomammillary nucleus (TMN) neurons using a conventional whole-cell patch clamp technique. Adenosine (100 microM) reversibly decreased mIPSC frequency without affecting the current amplitude, indicating that adenosine acts presynaptically to decrease the probability of spontaneous GABA release. The adenosine action on GABAergic mIPSC frequency was completely blocked by 1 microM DPCPX, a selective A(1) receptor antagonist, and mimicked by 1 microM CPA, a selective A(1) receptor agonist. This suggests that presynaptic A(1) receptors were responsible for the adenosine-mediated inhibition of GABAergic mIPSC frequency. CPA still decreased GABAergic mIPSC frequency even either in the presence of 200 microM Cd(2+), a general voltage-dependent Ca(2+) channel blocker, or in the Ca(2+)-free external solution. However, the inhibitory effect of CPA on GABAergic mIPSC frequency was completely occluded by 1 mM Ba(2+), a G protein coupled inwardly rectifying K(+) (GIRK) channel blocker. In addition, the CPA-induced decrease in mIPSC frequency was completely occluded by either 100 microM SQ22536, an adenylyl cyclase (AC) inhibitor, or 1 muM KT5720, a specific protein kinase A (PKA) inhibitor. The results suggest that the activation of presynaptic A(1) receptors decreases spontaneous GABAergic transmission onto TMN neurons via the modulation of GIRK channels as well as the AC/cAMP/PKA signal transduction pathway. This adenosine A(1) receptor-mediated modulation of GABAergic transmission onto TMN neurons may play an important role in the fine modulation of the excitability of TMN histaminergic neurons as well as the regulation of sleep-wakefulness. PMID- 18397366 TI - A novel intracellular role of matrix metalloproteinase-3 during apoptosis of dopaminergic cells. AB - We have previously demonstrated that the active form of matrix metalloproteinase 3 (actMMP-3) is released from dopamine(DA)rgic neurons undergoing apoptosis. Herein, whether actMMP-3 might be generated intracellularly, and if so, whether it is involved in apoptosis of DArgic neurons itself was investigated in primary cultured DArgic neurons of wild-type, MMP-3 knockout animals, and CATH.a cells. During apoptosis, gene expression of MMP-3 is induced, specifically among the various classes of MMPs, generating the proform (55 kDa) which is subsequently cleaved to the catalytically active actMMP-3 (48 kDa) involving a serine protease. Intracellular actMMP-3 activity is directly linked to apoptotic signaling in DArgic cells: (i) Pharmacologic inhibition of enzymatic activity, repression of gene expression by siRNA, and gene deficiency all lead to protection; (ii) pharmacologic inhibition causes attenuation of DNA fragmentation and caspase 3 activation, the indices of apoptosis; and (iii) inhibition of the pro-apoptotic enzyme c-Jun N-terminal protein kinase leads to repression of MMP-3 induction. Under the cell stress condition, MMP-3 is released as actMMP-3 rather than the proform (proMMP-3), and catalytically active MMP-3 added to the medium does not cause cell death. Thus, actMMP-3 seems to have a novel intracellular role in apoptotic DArgic cells and this finding provides an insight into the pathogenesis of Parkinson's disease. PMID- 18397367 TI - Characterization of PINK1 processing, stability, and subcellular localization. AB - Mutations found in PTEN-induced putative kinase 1 (PINK1), a putative mitochondrial serine/threonine kinase of unknown function, have been linked to autosomal recessive Parkinson's disease. It is suggested that mutations can cause a loss of PINK1 kinase activity and eventually lead to mitochondrial dysfunction. In this report, we examined the subcellular localization of PINK1 and the dynamic kinetics of PINK1 processing and degradation. We also identified cytosolic chaperone heat-shock protein 90 (Hsp90) as an interacting protein of PINK1 by PINK1 co-immunoprecipitation. Immunofluorescence of PINK1 protein and mitochondrial isolation show that the precursor form of PINK1 translocates to the mitochondria and is processed into two cleaved forms of PINK1, which in turn localize more to the cytosolic than mitochondrial fraction. The cleavage does not occur and the uncleaved precursor stays associated with the mitochondria when the mitochondrial membrane potential is disrupted. Metabolic labeling analyses show that the PINK1 processing is rapid and the levels of cleaved forms are tightly regulated. Furthermore, cleaved forms of PINK1 are stabilized by Hsp90 interaction as the loss of Hsp90 activity decreases PINK1 level after mitochondrial processing. Lastly, we also find that cleaved forms of PINK1 are degraded by the proteasome, which is uncommon for mitochondrial proteins. Our findings support a dual subcellular localization, implying that PINK1 can reside in the mitochondria and the cytosol. This raises intriguing functional roles that bridge these two cellular compartments. PMID- 18397369 TI - Etazolate, a neuroprotective drug linking GABA(A) receptor pharmacology to amyloid precursor protein processing. AB - Pharmacological modulation of the GABA(A) receptor has gained increasing attention as a potential treatment for central processes affected in Alzheimer disease (AD), including neuronal survival and cognition. The proteolytic cleavage of the amyloid precursor protein (APP) through the alpha-secretase pathway decreases in AD, concurrent with cognitive impairment. This APP cleavage occurs within the beta-amyloid peptide (Abeta) sequence, precluding formation of amyloidogenic peptides and leading to the release of the soluble N-terminal APP fragment (sAPPalpha) which is neurotrophic and procognitive. In this study, we show that at nanomolar-low micromolar concentrations, etazolate, a selective GABA(A) receptor modulator, stimulates sAPPalpha production in rat cortical neurons and in guinea pig brains. Etazolate (20 nM-2 microM) dose-dependently protected rat cortical neurons against Abeta-induced toxicity. The neuroprotective effects of etazolate were fully blocked by GABA(A) receptor antagonists indicating that this neuroprotection was due to GABA(A) receptor signalling. Baclofen, a GABA(B) receptor agonist failed to inhibit the Abeta induced neuronal death. Furthermore, both pharmacological alpha-secretase pathway inhibition and sAPPalpha immunoneutralization approaches prevented etazolate neuroprotection against Abeta, indicating that etazolate exerts its neuroprotective effect via sAPPalpha induction. Our findings therefore indicate a relationship between GABA(A) receptor signalling, the alpha-secretase pathway and neuroprotection, documenting a new therapeutic approach for AD treatment. PMID- 18397370 TI - Functional characterization of two cold shock domain proteins from Oryza sativa. AB - Two novel rice cold shock domain (CSD) proteins were cloned and characterized under different stress treatments and during various stages of development. OsCSP1 and OsCSP2 (Oryza sativa CSD protein) encode putative proteins consisting of an N-terminal CSD and glycine-rich regions that are interspersed by 4 and 2 CX(2)CX(4)HX(4)C (CCHC) retroviral-like zinc fingers, respectively. In vivo functional analysis confirmed that OsCSPs can complement a cold-sensitive bacterial strain which lacks four endogenous cold shock proteins. In vitro ssDNA binding assays determined that recombinant OsCSPs are capable of functioning as nucleic acid-binding proteins. Both OsCSP transcripts are transiently up regulated in response to low-temperature stress and rapidly return to a basal level of gene expression. Protein blot analysis determined that OsCSPs are maintained at a constant level subsequent to a cold treatment lasting over a period of several days. Both the transcript and protein data are in sharp contrast to those previously obtained for winter wheat WCSP1. A time-coursed study through various stages of rice development confirmed that both OsCSP proteins and transcripts are highly accumulated in reproductive tissues and tissues which exhibit meristematic activity. PMID- 18397368 TI - Modulation of taste sensitivity by GLP-1 signaling. AB - In many sensory systems, stimulus sensitivity is dynamically modulated through mechanisms of peripheral adaptation, efferent input, or hormonal action. In this way, responses to sensory stimuli can be optimized in the context of both the environment and the physiological state of the animal. Although the gustatory system critically influences food preference, food intake and metabolic homeostasis, the mechanisms for modulating taste sensitivity are poorly understood. In this study, we report that glucagon-like peptide-1 (GLP-1) signaling in taste buds modulates taste sensitivity in behaving mice. We find that GLP-1 is produced in two distinct subsets of mammalian taste cells, while the GLP-1 receptor is expressed on adjacent intragemmal afferent nerve fibers. GLP-1 receptor knockout mice show dramatically reduced taste responses to sweeteners in behavioral assays, indicating that GLP-1 signaling normally acts to maintain or enhance sweet taste sensitivity. A modest increase in citric acid taste sensitivity in these knockout mice suggests GLP-1 signaling may modulate sour taste, as well. Together, these findings suggest a novel paracrine mechanism for the regulation of taste function. PMID- 18397371 TI - CRY1 inhibits COP1-mediated degradation of BIT1, a MYB transcription factor, to activate blue light-dependent gene expression in Arabidopsis. AB - Cryptochromes (CRY) are one of the two major classes of photoreceptors that perceive light stimuli in the UV-A to blue light region and they are involved in multiple aspects of plant growth and development. However, knowledge regarding their signaling transduction components and mechanisms remains limited. Here, we report that a MYB transcription factor Blue Insensitive Trait 1 (BIT1), plays an important role in controlling blue light responses. Hypocotyl growth responses indicate that BIT1 functions as a positive element in blue light signaling, since BIT1 antisense and knock-out lines show a reduced light response in blue light. BIT1 controls blue light-dependent expression of various genes such as PsbS, a member of the light-harvesting complex gene family. A transactivation assay showed that BIT1 regulates promoter activity of PsbS in a blue light-dependent manner and that it requires CRY1 for activation of the PsbS promoter. BIT1 undergoes degradation in darkness and CRY1 functions to stabilize BIT1 in a blue light-dependent manner. In contrast, COP1 binds to BIT1 and mediates its degradation. We propose that the PsbS promoter is activated in blue light via the blue light-dependent stabilization of BIT1 by CRY1, while in darkness BIT1 is degraded by COP1-mediated proteolysis. PMID- 18397372 TI - Direct profiling and imaging of plant metabolites in intact tissues by using colloidal graphite-assisted laser desorption ionization mass spectrometry. AB - Laser desorption/ionization (LDI)-based imaging mass spectrometry (MS) has been applied to several biological systems to obtain information about both the identities of the major chemical species and their localization. Colloidal graphite-assisted LDI (GALDI) MS imaging was introduced for the imaging of small molecules such as phospholipids, cerebrosides, oligosaccharides, flavonoids, and other secondary metabolites with high spatial homogeneity due to finely dispersed particles. Mass profiles and images of Arabidopsis thaliana have been recorded directly from various plant surfaces and cross sections. The main targeted metabolites were flavonoids and cuticular waxes, both of which are important in many aspects of functional genomics, proteomics, and metabolomics. The mass spectral profiles revealed tissue-specific accumulation of flavonoids in flowers and petals. In addition, many other location-specific ions were observed. The location and the degree of light-induced accumulation of flavonoids in stem sections were successfully probed by GALDI MS. PMID- 18397373 TI - Regulation of root-wave response by extra large and conventional G proteins in Arabidopsis thaliana. AB - Heterotrimeric G proteins composed of alpha, beta and gamma subunits regulate a number of fundamental processes concerned with growth and development in plants. In addition to the canonical heterotrimeric G proteins, plants also contain a small family of extra large G proteins (XLGs) that show significant similarity to the G-protein alpha subunit in their C-terminal regions. In this paper we show that one of the three XLG genes, XLG3, and the Gbeta subunit (AGB1) of the Arabidopsis G-protein heterotrimer are specifically involved in the regulation of a subset of root morphological and growth responses. Based on analysis of T-DNA insertional mutant phenotypes, XLG3 and AGB1 each positively regulate root waving and root skewing. Since these responses are regulated by physical as well as physiological cues, we assessed the roles of AGB1 and XLG3 in gravitropism, thigmotropism and hormonal responses. Our data show that mutants lacking either XLG3 or AGB1 genes are hypersensitive to ethylene and show growth responses consistent with alterations in auxin transport, while maintaining an essentially wild-type response to the physical cues of gravity and touch. These results suggest that XLG3 and AGB1 proteins regulate the hormonal determinants of root waving and root-skewing responses in plants and possibly interact in a tissue specific or signal-specific manner. Because plants harboring knockout mutations in the Galpha subunit gene, GPA1, exhibit wild-type root waving and skewing, our results may indicate that the AGB1 subunit functions in these processes without formation of a classic Galphabetagamma heterotrimer. PMID- 18397374 TI - A tomato HD-Zip homeobox protein, LeHB-1, plays an important role in floral organogenesis and ripening. AB - Ethylene is required for climacteric fruit ripening. Inhibition of ethylene biosynthesis genes, 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase, prevents or delays ripening, but it is not known how these genes are modulated during normal development. LeHB-1, a previously uncharacterized tomato homeobox protein, was shown by gel retardation assay to interact with the promoter of LeACO1, an ACC oxidase gene expressed during ripening. Inhibition of LeHB-1 mRNA accumulation in tomato fruit, using virus-induced gene silencing, greatly reduced LeACO1 mRNA levels, and inhibited ripening. Conversely, ectopic overexpression of LeHB-1 by viral delivery to developing flowers elsewhere on injected plants triggered altered floral organ morphology, including production of multiple flowers within one sepal whorl, fusion of sepals and petals, and conversion of sepals into carpel-like structures that grew into fruits and ripened. Our findings suggest that LeHB-1 is not only involved in the control of ripening but also plays a critical role in floral organogenesis. PMID- 18397375 TI - Mixed-linkage beta-glucan : xyloglucan endotransglucosylase, a novel wall remodelling enzyme from Equisetum (horsetails) and charophytic algae. AB - Mixed-linkage (1-->3,1-->4)-beta-d-glucan (MLG), a hemicellulose long thought to be confined to certain Poales, was recently also found in Equisetum; xyloglucan occurs in all land plants. We now report that Equisetum possesses MLG:xyloglucan endotransglucosylase (MXE), which is a unique enzyme that grafts MLG to xyloglucan oligosaccharides (e.g. the heptasaccharide XXXGol). MXE occurs in all Equisetum species tested (Equisetum arvense, Equisetum fluviatile, Equisetum hyemale, Equisetum scirpoides, Equisetum telmateia and Equisetum variegatum), sometimes exceeding xyloglucan endotransglucosylase (XET) activity. Charophytic algae, especially Coleochaete, also possess MXE, which may therefore have been a primordial feature of plant cell walls. However, MXE was negligible in XET-rich extracts from grasses, dicotyledons, ferns, Selaginella and bryophytes. This and the following four additional observations indicate that MXE activity is not the result of a conventional xyloglucan endotransglucosylase/hydrolase (XTH): (i) XET, but not MXE, activity correlates with the reaction rate on water-soluble cellulose acetate, hydroxyethylcellulose and carboxymethylcellulose, (ii) MXE and XET activities peak in old and young Equisetum stems, respectively, (iii) MXE has a higher affinity for XXXGol (K(m) approximately 4 microM) than any known XTH, (iv) MXE and XET activities differ in their oligosaccharide acceptor-substrate preferences. High-molecular-weight (M(r)) xyloglucan strongly competes with [(3)H]XXXGol as the acceptor-substrate of MXE, whereas MLG oligosaccharides are poor acceptor-substrates. Thus, MLG-to-xyloglucan grafting appears to be the favoured activity of MXE. In conclusion, Equisetum has evolved MLG plus MXE, potentially a unique cell wall remodelling mechanism. The prominence of MXE in mature stems suggests a strengthening/repairing role. We propose that cereals, which possess MLG but lack MXE, might be engineered to express this Equisetum enzyme, thereby enhancing the crop mechanical properties. PMID- 18397376 TI - RLM3, a TIR domain encoding gene involved in broad-range immunity of Arabidopsis to necrotrophic fungal pathogens. AB - Here, we describe the rapid cloning of a plant gene, Leptosphaeria maculans 3 (RLM3(Col)), which encodes a putative Toll interleukin-1 receptor-nucleotide binding (TIR-NB) class protein, which is involved in defence against the fungal pathogen L. maculans and against three other necrotrophic fungi. We have, through microarray-based case control bulk segregant comparisons of transcriptomes in pools of Col-0 x An-1 progeny, identified the absence of a locus that causes susceptibility in An-1. The significance of this locus on chromosome 4 for L. maculans resistance was supported by PCR-based mapping, and denoted resistance to RLM3(Col). Differential susceptible phenotypes in four independent T-DNA insertion lines support the hypothesis that At4g16990 is required for RLM3(Col) function. The mutants in RLM3(Col) also exhibited an enhanced susceptibility to Botrytis cinerea, Alternaria brassicicola and Alternaria brassicae. Complementations of An-1 and T-DNA mutants using overexpression of a short transcript lacking the NB-ARC domain, or a genomic clone, restored resistance to all necrotrophic fungi. The elevated expression of RLM3(Col) on B. cinerea susceptible mutants further suggested convergence in signalling and gene regulation between defence against B. cinerea and L. maculans. In the case of L. maculans, RLM3(Col) is required for efficient callose deposition downstream of RLM1(Col). PMID- 18397377 TI - Cytokinins negatively regulate the root iron uptake machinery in Arabidopsis through a growth-dependent pathway. AB - Plants display a number of biochemical and developmental responses to low iron availability in order to increase iron uptake from the soil. The ferric-chelate reductase FRO2 and the ferrous iron transporter IRT1 control iron entry from the soil into the root epidermis. In Arabidopsis, expression of IRT1 and FRO2 is tightly controlled to maintain iron homeostasis, and involves local and long distance signals, as well as transcriptional and post-transcriptional events. FIT encodes a putative basic helix-loop-helix (bHLH) transcription factor that regulates iron uptake responses in Arabidopsis. Here, we uncover a new regulation of the root iron uptake genes. We show that IRT1, FRO2 and FIT are repressed by the exogenous addition of cytokinins (CKs), and that this repression acts at the level of transcript accumulation, and depends on the AHK3 and CRE1 CK receptors. The CKs and iron-deficiency signals act through distinct pathways to regulate the soil iron uptake genes, as (i) CK repression is independent of the iron status, (ii) IRT1 and FRO2 downregulation is unchanged in a fit loss-of-function mutant, indicating that FIT does not mediate CK repression, and (iii) the iron-regulated genes AtNRAMP3 and AtNRAMP4 are not downregulated by CKs. We show that root growth-inhibitory conditions, such as abiotic stresses (mannitol, NaCl) and hormonal treatments (auxin, abscissic acid), repress the iron starvation response genes. We propose that CKs control the root iron uptake machinery through a root growth dependent pathway in order to adapt nutrient uptake to the demand of the plant. PMID- 18397378 TI - Plant virus infection-induced persistent host gene downregulation in systemically infected leaves. AB - Understanding of virus infection-induced alterations in host plant gene expression and metabolism leading to the development of virus disease symptoms is both scientifically and economically important. Here, we show that viruses belonging to various RNA virus families are able to induce efficient host gene mRNA downregulation (shut-off) in systemically infected leaves. We demonstrate that the host gene mRNA shut-off overlaps spatially with virus-occupied sectors, indicating the direct role of virus accumulation in this phenomenon. The establishment of shut-off was not directly connected to active viral replication or the RNA-silencing machinery. Importantly, the induced shut-off phenomenon persisted for several weeks, resulting in severe deficiency of mRNA for important housekeeping genes in the infected plants. Interestingly, we found that some other RNA viruses do not induce or only slightly induce the shut-off phenomenon for the same set of genes, implicating genetic determination in this process. Nuclear run-on experiments suggest that plant viruses, similarly to animal viruses, mediate suppression of host mRNA synthesis in the nucleus. By investigating various host-virus interactions, we revealed a correlation between the intensity of the shut-off phenomenon and the severity of disease symptoms. Our data suggest that efficient and persistent downregulation of host genes may be an important component of symptom development in certain host-virus interactions. PMID- 18397379 TI - Defective in Tapetal development and function 1 is essential for anther development and tapetal function for microspore maturation in Arabidopsis. AB - In Arabidopsis, the tapetum plays important roles in anther development by providing enzymes for callose dissolution and materials for pollen-wall formation, and by supplying nutrients for pollen development. Here, we report the identification and characterization of a male-sterile mutant, defective in tapetal development and function 1 (tdf1), that exhibits irregular division and dysfunction of the tapetum. The TDF1 gene was characterized using a map-based cloning strategy, and was confirmed by genetic complementation. It encodes a putative R2R3 MYB transcription factor, and is highly expressed in the tapetum, meiocytes and microspores during anther development. Callose staining and gene expression analysis suggested that TDF1 may be a key component in controlling callose dissolution. Semi-quantitative and quantitative RT-PCR analysis showed that TDF1 acts downstream of DYT1 and upstream of AMS and AtMYB103 in the transcriptional regulatory networks that regulate tapetal development. In conclusion, our results show that TDF1 plays a vital role in tapetal differentiation and function. PMID- 18397380 TI - Alcohol trait and transcriptional genomic analysis of C57BL/6 substrains. AB - C57BL/6 inbred mice have been widely used as research models; however, widespread demand has led to the creation of several B6 substrains with markedly different phenotypes. In this study, we report that two substrains of C57BL/6 mice, C57BL/6J (B6J) and C57BL/6NCrl (B6C), separated over 50 years ago at two different breeding facilities differ significantly in alcohol consumption and alcohol preference. The genomes of these two substrains are estimated to differ by only 1-2% of all gene loci, providing a unique opportunity to extract particular expression signatures between these substrains that are associated with quantifiable behavioral differences. Expression profiling of the cortex and striatum, hippocampus, cerebellum and the ventral brain region from alcohol-naive B6C and B6J mice showed intervals on three chromosomes that are enriched in clusters of coregulated transcripts significantly divergent between the substrains. Additional analysis identified two genomic regions containing putative copy number differences between the substrains. One such region on chromosome 14 contained an estimated 3n copy number in the B6J genome compared with B6C. Within this interval, a gene of unknown function, D14Ertd449e, was found to be both associated with alcohol preference and vary in copy number across several inbred strain lineages. H2afz, Psen1, Wdfy1 and Clu were also identified as candidate genes that may be involved in influencing alcohol consumption. PMID- 18397381 TI - Loss of RAB-3/A in Caenorhabditis elegans and the mouse affects behavioral response to ethanol. AB - The mechanisms by which ethanol induces changes in behavior are not well understood. Here, we show that Caenorhabditis elegans loss-of-function mutations in the synaptic vesicle-associated RAB-3 protein and its guanosine triphosphate exchange factor AEX-3 confer resistance to the acute locomotor effects of ethanol. Similarly, mice lacking one or both copies of Rab3A are resistant to the ataxic and sedative effects of ethanol, and Rab3A haploinsufficiency increases voluntary ethanol consumption. These data suggest a conserved role of RAB-3 /RAB3A-regulated neurotransmitter release in ethanol-related behaviors. PMID- 18397382 TI - Nramp1 expression by dendritic cells modulates inflammatory responses during Salmonella Typhimurium infection. AB - Host resistance against Salmonella enterica serovar Typhimurium (S. Typhimurium) is mediated by natural resistance-associated macrophage protein 1 (Nramp1/Slc11a1). Nramp1 is critical to host defence, as mice lacking Nramp1 fail to control bacterial replication and succumb to low doses of S. Typhimurium. Despite this crucial role, the mechanisms underlying Nramp1's protective effects are unclear. Dendritic cells (DCs) that sample the intestinal lumen are among the first cells encountered by S. Typhimurium following oral infection and act as a conduit for S. Typhimurium to cross the intestinal epithelial barrier. We report that DCs, including intestinal, splenic and bone marrow-derived DCs (BMDCs), express Nramp1 protein. In the small intestine, Nramp1 expression is greater in a subset of DCs (CD11c(+)CD103(-)) characterized by the elevated expression of pro inflammatory cytokines in response to bacterial products. While Nramp1 expression did not affect S. Typhimurium replication in BMDCs, infected Nramp1+/+ BMDCs and intestinal CD11c(+)CD103(-) DCs secreted more inflammatory cytokines (IL-6, IL-12 and TNF-alpha) than Nramp1-/-, suggesting that Nramp1 expression may promote a more rapid inflammatory response following infection. Collectively, these findings reveal a new role for DCs and Nramp1 in modulating the host inflammatory response to S. Typhimurium. PMID- 18397383 TI - The role of beta2 integrins and lipopolysaccharide-binding protein in the phagocytosis of dead Neisseria meningitidis. AB - Phagocytosis of microbial pathogens is essential for the host immune response to infection. Our previous work has shown that lipooligosaccharide (LOS) expression on the surface of Neisseria meningitidis (Nm) is essential for phagocytosis, but the receptor involved remained unclear. In this study, we show that human CR3 (CD11b/CD18) and CR4 (CD11c/CD18) are phagocytic receptors for Nm as illustrated by the capacity of CR3- and CR4-transfected Chinese hamster ovary (CHO) cells to facilitate Nm uptake. A CR3-signalling mutant failed to internalize Nm, showing that the ability of CR3 to signal is essential for phagocytosis. Internalization of Nm by CR3-transfected CHO cells could be inhibited by the presence of CR3 specific antibodies. Furthermore, dendritic cells from leukocyte adhesion deficiency-1 patients, who have diminished expression of beta2 integrins, showed markedly reduced phagocytosis of Nm. The CR3-mediated phagocytosis required the presence of lipopolysaccharide-binding protein (LBP). Furthermore, the expression of LOS by Nm was essential for LBP binding and phagocytosis via CR3. These results reveal a critical role of CR3 and LBP in the phagocytosis of Nm and provide important insights into the initial interaction meningococci have with the immune system. PMID- 18397384 TI - Oestrogen receptors mediate oestrogen-induced increases in post-exercise rat skeletal muscle satellite cells. AB - AIM: Our laboratory recently demonstrated that increases in post-exercise muscle satellite cell numbers are augmented by oestrogen. We investigated whether muscle oestrogen receptors (ORs) mediate this effect through administration of an OR antagonist, ICI 182,780. METHODS: Ovariectomized female rats were divided into three groups: sham, oestrogen (0.25 mg pellet) and oestrogen plus OR blocker (ICI 182,780). Each group was divided into control and exercised groups. ICI 182,780 (5 mg kg(-1) sc) was administered 1 day prior to and 6 days following oestrogen pellet implantation. After 8 days of oestrogen exposure, animals ran downhill for 90 min (17 m min(-1), -13.5 degrees grade) on a treadmill. Soleus and white vastus muscles were removed 24 and 72 h post-exercise and immunostained for total (Pax7), activated (MyoD) and proliferating (BrdU) satellite cells. Muscle damage was indirectly assessed by measuring beta-glucuronidase activity. Two markers (His48 and ED1) of leucocyte infiltration were also examined. RESULTS: beta Glucuronidase activities and His48+ and ED1+ leucocytes increased post-exercise, and these increases were attenuated with oestrogen. ICI 182,780 did not influence the attenuating effect of oestrogen on leucocyte infiltration or beta glucuronidase activities in muscle. Total (Pax7+), activated (MyoD+) and proliferating (BrdU+) satellite cells increased post-exercise, and these increases were augmented with oestrogen. Interestingly, ICI 182,780 abolished both exercise- and oestrogen-mediated increases in these satellite cell markers. CONCLUSION: Oestrogen may augment increases in muscle satellite cells following exercise through OR-mediated mechanisms; furthermore, the attenuation of post exercise muscle damage and leucocyte infiltration by oestrogen appears to be a non-OR-mediated process. PMID- 18397385 TI - National mortality following upper gastrointestinal or cardiovascular events in older veterans with recent nonsteroidal anti-inflammatory drug use. AB - BACKGROUND: Upper gastrointestinal events (UGIE), myocardial infarction (MI) and cerebrovascular accident (CVA) are known morbidities among recent NSAID users. AIM: To assess all-cause mortality following UGIE, MI or CVA among recent NSAID users. METHODS: Veterans >65 prescribed an NSAID at any Veterans Affairs (VA) facility were identified using prescription fill data and their records linked to a merged VA-Medicare database. Each person-day was assessed for NSAID, coxib or proton pump inhibitor (PPI) exposure. Incidence density ratios and hazard rates of death were calculated following UGIE, MI and CVA adjusting for demographics, co-morbidity, prescription channeling, geographic location and pharmacological covariates. RESULTS: Among 474 495 patients [97.8% male; 85.3% white; 73.9 years (s.d. 5.6)], death followed at a rate of 5.5 per 1000 person-years (95% CI: 5.4 5.6) post-UGIE, 17.7 per 1000 person-years (95% CI: 17.5-17.9) post-MI and 21.8 per 1000 person-years (95% CI: 21.6-22.0) post-CVA. CVA was associated with greatest risk of death [hazard ratio (HR) 12.4; 95% CI: 10.9-14.3] followed by MI (HR 10.7; 95% CI: 9.2-11.6) and UGIE (HR 3.3; 95% CI: 2.8-3.9). Predictors of mortality were advancing age and co-morbidity, increased use of coxibs and failure to ensure adequate gastroprotection. CONCLUSION: Among elderly veterans with recent NSAID use, an UGIE, MI or CVA is a clinically relevant premorbid event. PMID- 18397386 TI - Clinical trial: exposure to ribavirin predicts EVR and SVR in patients with HCV genotype 1 infection treated with peginterferon alpha-2a plus ribavirin. AB - BACKGROUND: The impact of reduced drug exposure on outcomes in patients with chronic hepatitis C has not been determined in routine clinical practice. AIM: To examine the impact of exposure to peginterferon alpha-2a and ribavirin on early virological response (EVR) and sustained virological response (SVR) in treatment naive patients with HCV genotype 1 infection enrolled in a large expanded access programme. METHODS: Eight hundred and ninety-one patients treated for 48 weeks with an initial ribavirin dose of 800 or 1000/1200 mg/day were evaluated. Ribavirin 1000 mg/day (<75 kg) or 1200 mg/day (>or=75 kg) and peginterferon alpha 2a 180 microg/week were considered optimal. The impact of reduced drug exposure (expressed as a percentage of optimal) on EVR and SVR was evaluated. RESULTS: Mean ribavirin exposure in week 0-12 was 70% and 96% in patients assigned to ribavirin 800 and 1000/1200 mg/day, respectively. EVR and SVR rates were lower in patients assigned to ribavirin 800 than 1000/1200 mg/day (EVR, 75% vs. 84%, respectively, P < 0.001; SVR, 45% vs. 54%, respectively, P = 0.011). Furthermore, there was a strong correlation between achievement of EVR and SVR and ribavirin dose over the first 12 weeks expressed either as absolute dose or proportion of optimal dose received (P < 0.001 for both). CONCLUSIONS: Ribavirin exposure to week 12 is significantly associated with EVR and SVR in genotype 1 patients. Maintenance of an optimal ribavirin dose is the most important modifiable factor during combination therapy for chronic hepatitis C. PMID- 18397387 TI - Review article: epidemiology, pathogenesis and potential treatments of paediatric non-alcoholic fatty liver disease. AB - BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the most common cause of paediatric liver disease. Similar to NAFLD in adults, NAFLD in children is associated with obesity and insulin resistance and requires liver histology for diagnosis and staging. However, significant histological differences exist between adult and paediatric NAFLD to warrant caution in extrapolation of adult data. AIM: To review the available data on the epidemiology, pathogenesis, diagnosis and treatment of paediatric NAFLD. METHODS: Relevant articles were identified by Medline searches using the keywords: nonalcoholic fatty liver disease, steatohepatitis, obesity and children. RESULTS: The rise in childhood obesity has been accompanied by an increase in paediatric NAFLD. Age, gender and race/ethnicity are significant determinants of risk, and sex hormones, insulin sensitivity and adipocytokines are implicated in the pathogenesis of paediatric NAFLD. There is no consensus for treatment of NAFLD; however, data suggest that diet, exercise and some pharmacological therapies may be of benefit. CONCLUSIONS: To evaluate and effectively treat paediatric NAFLD, the pathophysiology and natural history of the disease should be clarified and non-invasive methods for screening, diagnosis, and longitudinal assessment developed. Randomized, controlled, double-blind trials of pharmacological therapies in children with biopsy-proven disease are necessary. PMID- 18397389 TI - Transient elastography accurately predicts presence of significant portal hypertension in patients with chronic liver disease. AB - BACKGROUND: Hepatic venous pressure gradient (HVPG) is a prognostic marker in patients with cirrhosis. Transient elastography measures liver stiffness (LS). AIM: To assess the correlation between LS and HVPG and to investigate the performance of transient elastography for the diagnosis of significant portal hypertension (PHT). METHODS: Liver stiffness was measured by Fibroscan in 150 consecutive patients who underwent a liver biopsy with haemodynamic measurements. Usual clinical and biological data were collected. Significant PHT was defined as a HVPG > or = 10 mmHg. RESULTS: Hepatic venous pressure gradient was found to be > or = 10 mmHg in 76 patients. Cirrhosis was diagnosed in 89 patients. HVPG was found to be correlated with: LS (rho = 0.858; P < 0.001) and inversely correlated with prothrombin index (rho = -0.718; P < 0.001). Regarding significant PHT, AUROC for LS and prothrombin index were 0.945 [0.904-0.987] and 0.892 [0.837 0.947] respectively. The cut-off value of 21 kPa accurately predicted significant PHT in 92% of the 144 patients for whom LS was successful. CONCLUSION: Liver stiffness measurement is correlated with HVPG and transient elastography identifies patients with significant PHT. PMID- 18397390 TI - Excellent prognosis and prevalence of HCV infection of primary hepatic and splenic non-Hodgkin's lymphoma. AB - BACKGROUND: Primary Hepatic (PHL) and Primary Splenic (PSL) non-Hodgkin's Lymphoma are rare entities. Small series of PHL and PSL have been reported, suggesting a non-fortuitous association with Hepatitis C Virus (HCV) infection. The prognosis is believed to be dismal, with early recurrence and short survival. PATIENTS: We retrospectively reviewed all PHL and PSL patients diagnosed at our institution between 1990 and 2005. RESULTS: Twenty-five adult patients were identified, six with PHL and 19 with PSL. Twenty-four patients had a B-cell lymphoma, defined as Diffuse Large B-cell lymphoma in 18. The prevalence of HCV infection was 68% among PSL and 66% among PHL. Combination chemotherapy was the mainstay of treatment for PHL and PSL; all but one patient with PSL underwent splenectomy before chemotherapy. Complete remission was achieved in all the cases after frontline therapy; only four patients relapsed but responded to additional chemotherapy courses. Most patients presented with aggressive histological subtypes; 92% were alive at a median follow up of 79 months. HCV infection did not appear to influence the results of therapy. CONCLUSION: Our study confirms the rarity of PHL and PSL, shows a high prevalence of HCV infection, and demonstrates that the outcome of patients with PHL and PSL may be favourable. PMID- 18397391 TI - Is Blastocystis hominis a new etiologic factor or a coincidence in iron deficiency anemia? AB - Iron deficiency anemia (IDA) is a frequent health problem. Gut parasites such as N. americanus and A. duodenale are known to cause blood loss, but the role of Blastocystis hominis is uncertain. In this study, 212 patients (193 female, 19 male, mean age 41 SD 15 yrs) with IDA were enrolled and 90 persons without IDA (78 female, 12 male, mean age 45 SD 17 yrs). Microscopic examination of stools for B. hominis using the native lugol method was done three times on each subject. If any specimen contained five or more cysts per x400 field, the person was considered positive. B. hominis was found in 48 out of 212 subjects with IDA (22.6%) and in five of 90 (5.6%) subjects without IDA. This difference is highly statistically significant (P < 0.001). Few subjects had other gut parasites and there was no statistical difference in the ir frequencies between IDA and non-IDA subjects. Blastocystis hominis may play a role in the development of IDA either on its own or in conjunction with some other agent. PMID- 18397392 TI - Eliminating lymphatic filariasis--the surveillance challenge. PMID- 18397393 TI - Relapses vs. reactions in multibacillary leprosy: proposal of new relapse criteria. AB - OBJECTIVES: To compare a new scoring system for multibacillary (MB) leprosy relapses, which combines time factor, risk factors and clinical presentation at relapse, to WHO criteria. METHODS: Data were collected on all relapses diagnosed between 1998 and 2004 at the Marie-Adelaide-Centre in Karachi, Pakistan, including case histories, clinical manifestations, follow-up, bacterial indices, treatment and contacts. For the diagnosis of MB relapses a simple scoring system was developed and validated on a data-set of mouse foot pads (MFP)-confirmed relapses (Leprosy Reviews, 76, 2005, 241). Its sensitivity was further evaluated in the Karachi relapse cohort. The P-value was calculated with McNemar's test with continuity correction. RESULTS: The new scoring system that combines time factor, risk factors and clinical presentation at relapse had a higher sensitivity in MFP-confirmed relapses than the WHO-criteria (95%vs. 65%, P < 0.01). The sensitivity of the scoring system was also significantly higher than the WHO criteria in the 57 cases of MB-relapses diagnosed in Karachi (72%vs. 54%, P < 0.05). CONCLUSIONS: This new simple scoring system for diagnosing MB-relapses in leprosy should be further validated in a prospective study to confirm its superior sensitivity and to evaluate the specificity of these criteria by using MFP-confirmation for patients presenting with signs of activity after treatment. PMID- 18397394 TI - Diversity of risk of mother-to-child HIV-1 transmission according to feeding practices, CD4 cell count, and haemoglobin concentration in a South African cohort. AB - OBJECTIVE: To estimate the probabilities of intrapartum and postpartum HIV-1 mother-to-child-transmission according to various feeding practices (formula feeding, exclusive breastfeeding, or mixed-feeding) and to other mother and infant covariates. METHODS: We used the promotion time model extended to multiple exposures to study the probability of infection attributable to each transmission occasion. Blood samples from 551 infants from Durban (South Africa) born to HIV-1 positive untreated mothers between 1995 and 1998 were sequentially tested until 15 months. RESULTS: The probability of infection attributable to in utero and intrapartum transmission was 21.88% (18.71-25.20) and was not significantly associated with the feeding practice. The probability of infection attributable to postnatal transmission through exclusive breastfeeding was negligible -0.7% (0 2.5) for 6 months of exclusive breastfeeding-- in comparison with that observed with mixed-feeding -6.15% (3.16-9.57) for 6 months of mixed-feeding. Maternal CD4 cell count and gestational age were significant predictors of intrapartum transmission probability while maternal CD4 cell count and maternal haemoglobin concentration were significant predictors of postpartum transmission probability. CONCLUSION: Decisions about appropriate infant feeding practices should take into account the difference in postpartum transmission risk between exclusive and mixed-feeding. Mixed-feeding should be all the more avoided that mothers have poor immunological statuses and low haemoglobin concentrations. PMID- 18397395 TI - Uptake of HIV voluntary counselling and testing services in rural Tanzania: implications for effective HIV prevention and equitable access to treatment. AB - OBJECTIVE: To describe the associations between socio-demographic, behavioural and clinical characteristics and the use of HIV voluntary counselling and testing (VCT) services among residents in a rural ward in Tanzania. METHODS: Eight thousand nine hundred and seventy participants from a community-based cohort were interviewed, provided blood for research HIV testing, and were offered VCT. Univariate and multivariate logistic regression was used to identify socio demographic, clinical and behavioural factors associated with VCT use. RESULTS: Although 31% (1246/3980) of men and 24% (1195/4990) of women expressed an interest in the service, only 12% of men and 7% of women subsequently completed VCT. Socio-demographic factors, such as marital status, area of residence, religion and ethnicity influenced VCT completion among males and females in different ways, while self-perceived risk of HIV, prior knowledge of VCT, and sex with a high-risk partner emerged as important predictors of VCT completion among both sexes. Among males only, those infected with HIV for 5 years or less tended to self-select for VCT compared to HIV-negatives (adjusted odds ratio = 1.43; 95% CI: 0.99-2.14). This contributed to a higher proportion of HIV-positive males knowing their status compared to HIV-positive females. CONCLUSIONS: In this setting, a disproportionate number of HIV-positive women are failing to learn their status, which has implications for equitable access to onward referral for care and treatment services. Evidence that some high-risk behaviours may prompt VCT use is encouraging, although further interventions are required to improve knowledge about HIV risk and the benefits of VCT. Targeted interventions are also needed to promote VCT uptake among married women and rural residents. PMID- 18397396 TI - Sleeping sickness in West Africa (1906-2006): changes in spatial repartition and lessons from the past. AB - OBJECTIVE: To review the geography and history of sleeping sickness (Human African trypanosomiasis; HAT) over the past 100 years in West Africa, to identify priority areas for sleeping sickness surveillance and areas where HAT no longer seems active. METHOD: History and geography of HAT were summarized based on a review of old reports and recent publications and on recent results obtained from medical surveys conducted in West Africa up to 2006. RESULTS/CONCLUSIONS: Active HAT foci seem to have moved from the North to the South. Endemic HAT presently appears to be limited to areas where annual rainfall exceeds 1200 mm, although the reasons for this remain unknown. There has also been a shift towards the south of the isohyets and of the northern distribution limit of tsetse. Currently, the most severely affected countries are Guinea and Ivory Coast, whereas the northern countries seem less affected. However, many parts of West Africa still lack information on HAT and remain to be investigated. Of particular interest are the consequences of the recent political crisis in Ivory Coast and the resulting massive population movements, given the possible consequences on HAT in neighbouring countries. PMID- 18397397 TI - Nutritional status and common infections in infants in Burkina Faso: interest of an 'overall morbidity score'. AB - OBJECTIVE: To study the overall impact of morbidity on the nutritional status of infants in a rural area of Burkina-Faso. METHOD: A cohort of children (n = 114) were monitored from age 6 months to 2 years in 30 villages in north-eastern Burkina Faso. Anthropometric measurements and the incidence of illness were recorded at 6, 9, 12, 15, 18 and 24 months. An overall morbidity score was constructed including the duration of common illnesses and their effect on appetite. Mixed linear regression models were used to analyse the relation between this score and both the weight-for-length (WFL) and the length-for-age (LFA) index. RESULTS: After adjustment for potential confounders, the morbidity score at time T was strongly correlated with the WFL index at time T (P < 0.0001) and weakly correlated with the LFA index at time T + 1 (P = 0.06). When the morbidity score was divided into classes, at time T the mean WFL z-score for children with a high morbidity score was 0.34 z-scores lower than the one for healthy children (P < 0.0001); at time T + 1, the difference in the mean LFA z scores between the same groups was 0.09 z-score (P = 0.02). However, similar results were obtained using simpler expressions of morbidity. Taken separately, diarrhoea or febrile illness had an effect on WFL (at time T), but only diarrhoea had an effect on LFA (at time T + 1). CONCLUSION: Morbidity was strongly correlated with weight loss in children in the short term and weakly correlated with a slowdown in growth. PMID- 18397398 TI - Use of proxy measures in estimating socioeconomic inequalities in malaria prevalence. AB - OBJECTIVES: To present and compare socioeconomic status (SES) rankings of households using consumption and an asset-based index as two alternative measures of SES; and to compare and evaluate the performance of these two measures in multivariate analyses of the socioeconomic gradient in malaria prevalence. METHODS: Data for the study come from a survey of 557 households in 25 study villages in Tanzania in 2004. Household SES was determined using consumption and an asset-based index calculated using Principal Components Analysis on a set of household variables. In multivariate analyses of malaria prevalence, we also used two other measures of disease prevalence: parasitaemia and self-report of malaria or fever in the 2 weeks before interview. RESULTS: Household rankings based on the two measures of SES differ substantially. In multivariate analyses, there was a statistically significant negative association between both measures of SES and parasitaemia but not between either measure of SES and self-reported malaria. Age of individual, use of a mosquito net, and wall construction were negatively and significantly associated with parasitaemia, whilst roof construction was positively associated with parasitaemia. Only age remained significant when malaria self-report was used as the measure of disease prevalence. CONCLUSIONS: An asset index is an effective alternative to consumption in measuring the socioeconomic gradient in malaria parasitaemia, but self-report may be an unreliable measure of malaria prevalence for this purpose. PMID- 18397399 TI - Setting up a national reference laboratory for Buruli ulcer: the case of Benin. AB - OBJECTIVE: To report the experience of Benin, where Buruli ulcer (BU) is endemic, in the implementation of diagnostic laboratory services. METHODS AND RESULTS: There has been a gradual introduction of biologic diagnostic activities for BU comprising (1) training of a laboratory technician in a highly experienced reference laboratory; (2) acquiring indispensable laboratory start-up materials; (3) progressive development of diagnostic laboratory activities; (4) regular external quality assessment with an experienced reference laboratory and (5) decentralization of activities to various clinical diagnostic and treatment centres for BU in Benin. CONCLUSION: Setting up a reference laboratory for BU is a continuous process, which necessitates motivated personnel and the cooperation of an experienced external reference laboratory. PMID- 18397400 TI - Primary health care supervision in developing countries. AB - OBJECTIVES: To (a) summarise opinion about what supervision of primary health care is by those advocating it; (b) compare these features with reports describing supervision in practice; and (c) to appraise the evidence of the effects of sector performance. METHODS: Systematic review. Reports were classified into three groups and summarised using appropriate methods: policy and opinion papers (narrative summary), descriptive studies (systematically summarised) and experimental or quasi-experimental studies (design and outcomes systematically summarised). Data presented as narrative summaries and tables. RESULTS: 74 reports were included. In eight policy and opinion papers, supervision was conceptualised as the link between the district and the peripheral health staff; it is important in performance and staff motivation; it often includes problem solving, reviewing records, and observing clinical practice; and is usually undertaken by visiting the supervisees place of work. In 54 descriptive studies, the setting was the primary health care (PHC) or specific services and programmes. Supervisor-supervisee dyads were generally district personnel supervising health facilities or lay health workers. Supervision mostly meant visiting supervisees, but also included meetings in the centre; it appeared to focus on administration and checking, sometimes with checklists. Problem solving, feedback and clinical supervision, training and consultation with the community were less commonly described in the descriptive studies. Supervision appears expensive from studies that have reported costs. In 12 quasi-experimental trials, supervision interventions generally showed small positive effects in some of the outcomes assessed. However, trial quality was mixed, and outcomes varied greatly between studies. CONCLUSIONS: Supervision is widely recommended, but is a complex intervention and implemented in different ways. There is some evidence of benefit on health care performance, but the studies are generally limited in the rigor and follow up is limited. Further research delineating what supervision consists of and evaluating it in the context of unbiased comparisons would guide the implementation of effective supervision as part of the management of PHC. PMID- 18397401 TI - Assessing the validity of health facility-based data on insecticide-treated bednet possession and use: comparison of data collected via health facility and household surveys--Lindi region and Rufiji district, Tanzania, 2005. AB - Objective To assess the validity of health facility (HF)-based data on bednet and insecticide-treated bednet possession and use by children <5 years old. Methods We compared estimates based on data collected via HF surveys of under-5s attending well-child visits (e.g. immunizations) and sick-child visits vs. representative household surveys (a 'gold standard' method for measuring insecticide-treated net coverage). In Lindi region, Tanzania, we collected contemporaneous data on 637 under-5s via a HF survey (444 well-child visits and 193 sick-child visits), and on 305 households with an under-5 (including 354 children) via a household survey. In Rufiji district, Tanzania, we collected contemporaneous data on 1433 under-5s via a HF survey (911 well-child visits and 522 sick-child visits), and on 328 households with an under-5 (including 455 children) via a household survey. Results Possession of bednets by households with an under-5 was similar using HF data and household data in both Lindi region and Rufiji district. However, reported use of bednets was significantly higher in HF data than household data in both Lindi and Rufiji, as was reported use of insecticide-treated bednets. HF-based data accurately estimated community-level bednet possession in households with an under-5, but overestimated community level bednet use by 9-35% and insecticide-treated bednet use by 15-21%. Conclusions Information bias rather than selection bias appears to be a key cause for the overestimation of bednet and insecticide-treated bednet use (e.g. social desirability bias: caretakers of under-5s attending health facilities might be more likely to report using bednets and insecticide-treated bednets). Additional studies of the validity, cost and utility of HF-based data to monitor insecticide treated bednet use are needed before recommending this monitoring strategy for widespread use. Overestimating insecticide-treated bednet use could lead to inappropriate public health actions and missed opportunities for achieving local and global public health goals. PMID- 18397402 TI - Clinical diagnosis of malaria and the risk of chloroquine self-medication in rural health centres in Burkina Faso. AB - OBJECTIVE: To assess the quality of healthcare workers' performance with regard to malaria diagnosis and treatment and to assess patients' self-medication with chloroquine (CQ) before and after presentation at a health centre. METHODS: In the rainy season 2004, in five rural dispensaries in Burkina Faso, we observed 1101 general outpatient consultations and re-examined all these patients. CQ whole blood concentrations of confirmed malaria cases were measured before and after treatment. RESULTS: The clinical diagnosis based on fever and/or a history of fever had a sensitivity of 75% and a specificity of 41% when compared to confirmed malaria (defined as an axillary temperature of >/=37.5 degrees C and/or a history of fever and parasites of any density in the blood smear). Few febrile children under 5 years of age were assessed for other diseases than malaria such as pneumonia. No antimalarial was prescribed for 1.3% of patients with the clinical diagnosis malaria and for 24% of confirmed cases, while 2% received an antimalarial drug prescription without the corresponding clinical diagnosis. CQ was overdosed in 22% of the prescriptions. Before and 2 weeks after consultation, 25% and 46% respectively of the patients with confirmed malaria had potentially toxic CQ concentrations. CONCLUSION: As long as artemisinin-based combination therapy remains unavailable or unaffordable for most people in rural areas of Burkina Faso, self-medication with and prescription of CQ are likely to continue despite increasing resistance. Apart from considering more pragmatic first-line regimens for malaria treatment such as the combination of sulfadoxine pyrimethamine with amodiaquine, more and better training on careful clinical management of febrile children including an appropriate consideration of other illnesses than malaria should be made available in the frame of the IMCI initiative in sub-Saharan Africa. PMID- 18397403 TI - Low carriage rate of group B streptococcus in pregnant women in Maputo, Mozambique. AB - The prevalence of group B streptococcus (GBS) carriage varies strongly with geographical region. A study was done to determine the prevalence of GBS in women in Maputo, Mozambique. The method used was a rectovaginal swab which was taken from women between 35 and 37 weeks of pregnancy who visited the clinic for antenatal consultation. GBS was cultured from 2 out of 113 samples, yielding a prevalence of 1.8% (95% Cl: 0.0-4.0). In conclusion, the prevalence of GBS carriage among pregnant women in Maputo, Mozambique was low. PMID- 18397404 TI - Knockdown resistance mutations (kdr) and insecticide susceptibility to DDT and pyrethroids in Anopheles gambiae from Equatorial Guinea. AB - OBJECTIVES: To determine the frequency of knockdown resistance (kdr) mutations in the malaria vector Anopheles gambiae s.s. from continental Equatorial Guinea; and to relate kdr genotypes with susceptibility to DDT and pyrethroid insecticides in this vector. METHODS: Female mosquitoes were collected in two villages, Miyobo and Ngonamanga, of mainland Equatorial Guinea. Insecticide susceptibility tests were performed following WHO procedures. Anopheles gambiae complex specimens were identified to species and molecular form by PCR. Genotyping of the kdr locus was performed by allele-specific PCR and direct sequencing in a subset of samples. RESULTS: Both M and S molecular forms of A. gambiae were found in Ngonamanga whereas only the S-form was identified in Miyobo. The two kdr mutations were detected in S-form samples of both villages, with a higher frequency of the kdr-e (Leu-1014-Ser) allele (Miyobo: 16%; Ngonamanga: 40%). The kdr-w (Leu-1014-Phe) mutation was also detected in 3% of the M-form. All individuals tested for pyrethroids were susceptible. A mortality rate of 86% was obtained for DDT. An overall kdr allele frequency (i.e. kdr-e + kdr-w) of 22% was detected in DDT resistant individuals, whereas susceptible individuals had a kdr frequency of 6%. CONCLUSION: The co-occurrence of both kdr mutations and reduced susceptibility to DDT found in A. gambiae highlights the importance of implementing efficient surveillance of insecticide resistance in Equatorial Guinea. PMID- 18397405 TI - Antibiotic use and health-seeking behaviour in an underprivileged area of Peru. AB - OBJECTIVE: To describe the health-seeking behaviour and use of antibiotics in the urban community of Yurimaguas in the Amazonian area of Peru. METHOD: Cross sectional survey of caregivers of 798 children aged 6-72 months by interview using a semi-structured questionnaire. Reported symptoms were classified as illnesses where antibiotics would or would not be recommended based on principles of the integrated management of childhood illnesses algorithm. RESULTS: Forty-one per cent of consultations were with health care professionals; 71% of antibiotics were obtained through the formal public health sector and prescribed mainly by medical doctors. All prescribed antibiotics were on the Peruvian essential drugs list. When prescribing, doctors and nurses hardly discriminated between illnesses where antibiotic treatment was or was not indicated; there was no significant difference in antibiotic prescribing rates between the two (doctors, P = 0.24; nurses, P = 0.32). Not all caregivers sought help for children with severe symptoms. CONCLUSION: Although most of the antibiotics were prescribed by doctors and nurses, they were commonly prescribed for illnesses where they were not indicated. The use of antibiotics needs to be rationalized, and barriers to health care must be overcome. PMID- 18397406 TI - Cysteinyl-leukotrienes in nasal lavage fluid in children with asthma. AB - The aim of this study was to investigate repeatability of cysteinyl-leukotrienes (cys-LT) measurements in nasal lavage fluid (NLF) and to determine if cys-LT levels in NLF are related to asthma severity in children. As a second outcome, we investigated if cys-LT in NLF reflect lower airway inflammation as assessed by exhaled NO measurement. To assess the repeatability of cys-LT measurements, two NLF samples were obtained from eight healthy controls 24 h apart. Sixty-nine asthmatic children (mean age; range: 12.8; 7.3-17.7 yr), which were grouped according to asthma severity were studied cross-sectionally on one occasion. Cys LT in NLF were analyzed using a specific enzyme immunoassay, exhaled NO, and pulmonary function parameters were measured. The coefficient of repeatability for the repeated cys-LT measurements was 1.45 pg/ml. Cys-LT levels in NLF differed significantly between asthma severity groups (p < 0.001): mild intermittent: [median (IQR)] 6.88 pg/ml (2.00-27.87); mild persistent: 21.09 pg/ml (4.50 84.67); and moderate persistent asthmatics: 36.41 pg/ml (11.03-118.40). Concentration of cys-LT in NLF and exhaled NO was positively correlated (r = 0.85; p < 0.001). In conclusion, concentration of cys-LT in NLF correlates with asthma severity in children and is related to lower airway inflammation. PMID- 18397407 TI - A methodology for measurements of nasal nitric oxide in children under 5 yr. AB - Measurements of nasal nitric oxide (nNO) may give insight into respiratory conditions in children aged under 5 yr but no methodology has been described for this age-group. The present study aimed to establish the methodology and reproducibility for measuring nNO during tidal breathing in young children and to relate nNO to allergic conditions. Children and siblings aged under 5 yr attending hospital clinics were enrolled. On-line nNO measurements were obtained during tidal breathing using a chemiluminescence analyser. To establish our methodology, nNO was measured over 3, 5 or 10 s NO plateaus and nNO was also measured from left and right nostrils. nNO was then compared between children with and without allergic conditions. The reproducibility of nNO measurements over 24 h was studied in a separate group of children. Eighty-three children participated in the methodological part of the study and nNO was successfully measured in 57 (69%), mean (s.d.) age 3.4 (1.1) years, 14 with allergic conditions. Neither NO plateau duration nor choice of nostril influenced nNO values. The mean (s.d.) nNO for non-atopic children was 208 (103) parts per billion (ppb) and for atopic children was 284 (122), p = 0.032. Nasal NO values were not related to ambient NO, gender and passive smoke exposure; there was a non-significant trend for nNO to be positively related to age. Nasal NO measurements were reproducible in the 21 children tested, mean difference 9.6 ppb (limits of agreement-127, 146). We report a methodology for nNO measurement in young children. Further work is now required to establish the clinical utility of nNO in this age-group. PMID- 18397408 TI - Cord blood hemopoietic progenitor profiles predict acute respiratory symptoms in infancy. AB - Atopy is characterized by eosinophilic inflammation associated with recruitment of eosinophil/basophil (Eo/B) progenitors. We have previously shown that Eo/B progenitor phenotypes are altered in cord blood (CB) in infants at high risk of atopy/asthma, and respond to maternal dietary intervention during pregnancy. As respiratory tract viral infections have been shown to induce wheeze in infancy, we investigated the relationship between CB progenitor function and phenotype and acute respiratory illness (ARI), specifically wheeze and fever. CB from 39 high risk infants was studied by flow cytometry for CD34(+) progenitor phenotype and by ex vivo Eo/B-colony forming unit (CFU) responses to cytokine stimulation in relation to ARI in the first year of life. A consistent relationship was observed between increased numbers of granulocyte/macrophage (GM)-colony-stimulating factor (CSF)- and IL-3-responsive Eo/B-CFU in CB and the frequency/characteristics of ARI during infancy. Comparable associations were found between ARI and CB IL-3R(+) and GM-CSFR(+)CD34(+) cell numbers. Conversely, a reciprocal decrease in the proportion of CB IL-5R(+) cells was found in relation to the clinical outcomes. The elevation of IL-3/GM-CSF-responsive Eo/B progenitors in high-risk infants in relation to ARI outcomes suggests a mechanism for the increased severity of inflammatory responses in these subjects following viral infection. PMID- 18397409 TI - Non-atopic asthma in children is related to maternal bronchial hyperreactivity. AB - Data on the pathogenic mechanisms underlying the development of non-atopic asthma in children are scarce. Our aim was to evaluate the association and compare the atopic status, pulmonary functions, bronchial hyperresponsiveness and serum total immunoglobulin E (IgE) levels of parents of atopic and non-atopic asthmatic children by using objective methods. Fifty-one asthmatic children aged 4-16 yr and their parents were included into the study. Initially the American Thoracic Society's Respiratory Disease questionnaire inquiring data on symptoms of asthma, rhinitis and past medical history was filled in. Afterwards, skin prick test with aeroallergens, pulmonary function and methacholine bronchial provocation tests and serum sampling for total IgE level determinations were carried out. Bronchial hyperresponsiveness to methacholine was significantly more common in the mothers of non-atopic children compared to those of atopic ones, although no significant difference was observed in the skin prick test reactivity, pulmonary function test parameters and serum IgE levels. Questionnaire data revealed that the presence of asthmatic symptoms such as wheezing and phlegm and doctor-diagnosed asthma were more common in the mothers of non-atopic children. Meanwhile, asthmatic symptoms were also found to be significantly more common in fathers of non-atopic children. Logistic regression analyses revealed that maternal PC(20) was the only predictive factor for the risk of displaying non-allergic asthma in children. The results demonstrate that among the risk factors studied, maternal bronchial hyperreactivity was associated with the development of asthma in non atopic children. PMID- 18397410 TI - Stressful life events and asthma in adolescents. AB - Recent studies suggest that psychosocial factors may contribute to asthma. We examined associations of stressful life events with asthma prevalence and morbidity among Chicago adolescents. Self-reported asthma, measures of asthma morbidity, and 15 life events were collected from 2026 seventh to ninth grade students from 34 Chicago Catholic schools as part of the International Study of Allergies and Asthma in Childhood in 1994-95. Life events were reported by 77% of adolescents and overall asthma prevalence was 15.5%. Stressful life events in adolescents were significantly related to both asthma and asthma morbidity. Odds of asthma was 1.44 (95% confidence interval [CI] = 1.07, 1.95) for those reporting two to three stressful events and 1.92 (95% CI = 1.41, 2.62) for subjects endorsing more than three stressful events. In adolescents with asthma, number of asthma symptoms (odds ratio [OR] for increase in one event = 1.16, 95% CI = 1.07, 1.27), asthma-related school absenteeism (OR = 1.17, 95% CI = 1.04, 1.32), physician visits for asthma (OR = 1.16, 95% CI = 1.04, 1.29), and hospitalization for asthma (OR = 1.20, 95% CI = 1.001, 1.44) were significantly associated with the number of stressful events, independent of home exposure to cigarette smoke and dampness, use of inhaled substances, and sociodemographic factors. While these results are not sufficient to assign causality in the relationship between stress and asthma, they are supported by a number of other studies and by plausible biologic mechanisms. Assessing and addressing the effects of stressful life events may be helpful in managing asthma in inner city adolescents. PMID- 18397411 TI - A survey of patients with self-reported severe food allergies in Japan. AB - Food allergies have increased in recent years in Japan. Details of causative foods, places where anaphylaxis developed, and other allergic factors remain unknown, and we investigated them. A'questionnaire survey for the prevention of food allergies' was conducted using a nationwide group of patients with food allergies. A total of 1383 patients from 878 families (including 319 patients who experienced anaphylaxis) provided valid answers to the questionnaire. The average age of the first anaphylactic attack was 3.20 +/- 6.327 yr. The most common allergens causing anaphylaxis were in order milk, eggs, wheat, peanuts, and soybeans, followed by sesame and buckwheat. The most common place where anaphylaxis developed was the patient's own home, followed by fast food restaurants, places visited, restaurants, and schools. In patients' own homes, fast food restaurants (buffet), places visited and schools, the most common allergens were milk, eggs, and wheat. In restaurants and accommodation facilities, eggs were the most common allergen followed by milk. As possible food allergies can cause anaphylaxis, it is necessary to provide precise information for consumers regarding packaged and processed foods. PMID- 18397412 TI - Cow's milk allergy: a diagnostic challenge. PMID- 18397414 TI - Food allergy in children suffering from atopic eczema. AB - Whilst the association between eczema and food allergy is well established, the role of dietary manipulation in children with eczema remains controversial. These case histories highlight the differing outcomes that dietary manipulation may have in an infant with early onset, severe eczema and an older child with milder eczema. Management strategies and the evidence to support them are presented, followed by a review of clinical recommendations. PMID- 18397418 TI - Balancing drug risk and benefit: toward refining the process of FDA decisions affecting patient care. AB - Several high-profile drug withdrawals for safety issues have brought into focus the FDA's process for approving drugs and monitoring adverse experiences with those agents after marketing has begun. Gastroenterologists and their patients have been affected adversely by removal from the marketplace of two licensed agents for irritable bowel syndrome (IBS): alosetron and tegaserod. The criteria used by the FDA for assessment of the risks and benefits of drugs used for functional bowel problems seem to be different than those used for the treatment of other conditions and have resulted in drastic limitation of access to these drugs rather than just warnings about risks as they are discovered. Decisions that affect the availability of drugs for patients with functional bowel disease should be discussed with clinicians who take care of those patients before going into effect. The absence of this sort of consultation leaves physicians with serious limitations on their abilities to take care of patients. PMID- 18397419 TI - How to do and interpret a rectal examination in gastroenterology. PMID- 18397420 TI - High prevalence of hepatitis C virus infection among immigrants from the former Soviet Union in the New York City metropolitan area: results of a community-based screening program. AB - BACKGROUND: Inadequate sterilization and reuse of medical equipment likely contributed to hepatitis C virus (HCV) transmission in the former Soviet Union (FSU). Although New York leads the nation in the number of immigrants from the FSU, the epidemiology of HCV infection has not been evaluated in this population. The aims of this study were to determine the prevalence of and risk factors for HCV infection among immigrants from the FSU in the New York metropolitan area. METHODS: We conducted a 3-day community-based HCV screening program in the two boroughs of the New York metropolitan area with the highest density of FSU immigrants (Brooklyn and Queens). Russian cable television was used to invite subjects to come in for free HCV testing. In the last 2 days of screening, each person also completed an HCV risk factor questionnaire. RESULTS: The overall prevalence of HCV seropositivity among the 283 subjects was 28.3% (95% confidence interval [CI] 23.0-33.5%). The prevalence of HCV infection was similar in men and women (30.3% vs 26.5%, P = 0.48) and was highest in subjects > or = 70 yr old (35.0%). HCV seropositivity was 11.1% in immigrants from Russia, 29.0% from Uzbekistan, 31.0% from the Ukraine, and 36.8% from other regions. Intramuscular injections (odds ratio 9.1, 95% CI 2.0-42.4) and blood transfusions (odds ratio 3.2, 95% CI 1.2-9.0) were the only variables that were significantly associated with HCV infection in the multivariable analysis. CONCLUSIONS: In this community based screening program we found a high prevalence of HCV infection among immigrants from the FSU, and these infections likely resulted from inadequately sterilized medical equipment and blood transfusions. Universal HCV testing should be strongly considered for all FSU immigrants. PMID- 18397425 TI - Percutaneous endoscopic gastrostomy in the dementia patient: helpful or hindering? AB - Percutaneous endoscopic gastrostomy (PEG) is a common technique performed worldwide. Recently, the use of PEG in the dementia patient with dysphagia for nutrition support has been called into question. Some reviews have reported no improvement in survival with PEG tubes in this population. Higaki and colleagues now present a large review of PEG tube placement in patients with dementia, which demonstrates a similar survival rate in patients without dementia receiving PEG tubes. The question of the utility of PEG tube placement for nutrition support in the dementia population requires an organized, prospective analysis to concretely answer the question. PMID- 18397427 TI - Resectability of pancreatic cancer and diabetes. PMID- 18397429 TI - The timing of colonoscopy preparation makes all the difference in the final outcome. PMID- 18397431 TI - An interleukin-6 (IL-6) receptor polymorphism affecting serum levels of IL-6 does not increase the risk of cholangiocarcinoma in primary sclerosing cholangitis. PMID- 18397432 TI - Abnormal Pap smears in women with Crohn's disease: is there a role for defensin deficiency? PMID- 18397433 TI - Hyperinsulinemic hypoglycemia after bariatric surgery. PMID- 18397435 TI - Cannabinoid hyperemesis. PMID- 18397437 TI - Diagnosis of cystic lymphangioma of the pancreas with endoscopic ultrasound guided fine needle aspiration. PMID- 18397438 TI - Dosage and drug interactions of domperidone. PMID- 18397439 TI - Spontaneous regression of hepatocellular carcinoma. PMID- 18397440 TI - Ascaris lumbricoides and iron deficiency anemia. PMID- 18397441 TI - Pancreatic cancer manifesting as liver metastases and eosinophillic leukemoid reaction: a case report and review of literature. PMID- 18397442 TI - Health care information technology in rural America: electronic medical record adoption status in meeting the national agenda. AB - Continuing is a national political drive for investments in health care information technology (HIT) that will allow the transformation of health care for quality improvement and cost reduction. Despite several initiatives by the federal government to spur this development, HIT implementation has been limited, particularly in the rural market. The status of technology use in the transformation effort is reviewed by examining electronic medical records (EMRs), analyzing the existing rural environment, identifying barriers and factors affecting their development and implementation, and recommending needed steps to make this transformation occur, particularly in rural communities. A review of the literature for HIT in rural settings indicates that very little progress has been made in the adoption and use of HIT in rural America. Financial barriers and a large number of HIT vendors offering different solutions present significant risks to rural health care providers wanting to invest in HIT. Although evidence in the literature has demonstrated benefits of adopting HIT such as EMRs, important technical, policy, organizational, and financial barriers still exist that prevent the implementation of these systems in rural settings. To expedite the spread of HIT in rural America, federal and state governments along with private payers, who are important beneficiaries of HIT, must make difficult decisions as to who pays for the investment in this technology, along with driving standards, simplifying approaches for reductions in risk, and creating a workable operational plan. PMID- 18397443 TI - The importance of location for tobacco cessation: rural-urban disparities in quit success in underserved West Virginia Counties. AB - CONTEXT: Adults who live in rural areas of the United States have among the highest smoking rates in the country. Rural populations, including Appalachian adults, have been historically underserved by tobacco control programs and policies and little is known about their effectiveness. PURPOSE: To examine the end-of-class quit success of participants in A Tobacco Cessation Project for Disadvantaged West Virginia Communities by place of residence (rural West Virginia and the urban area of Greater Charleston). METHODS: This collaborative program was implemented in 5 underserved rural counties in West Virginia and consisted of 4 intervention approaches: (1) a medical examination; (2) an 8 session educational and behavioral modification program; (3) an 8-week supply of pharmacotherapy; and (4) follow-up support group meetings. FINDINGS: Of the 725 program participants, 385 (53.1%) had successfully quit using tobacco at the last group cessation class they attended. Participants who lived in rural West Virginia counties had a lower end-of-class quit success rate than those who lived in the urban area of Greater Charleston (unadjusted odds ratio [OR]= 0.69, 95% confidence interval [CI]= 0.48, 0.99), even after taking into account other characteristics known to influence quit success (adjusted OR = 0.58, 95% CI = 0.35, 0.94). CONCLUSIONS: Tobacco control programs in rural West Virginia would do well to build upon the positive aspects of rural life while addressing the infrastructure and economic needs of the region. End-of-class quit success may usefully be viewed as a stage on the continuum of change toward long-term quit success. PMID- 18397444 TI - Understanding smoking cessation in rural communities. AB - CONTEXT: Rural communities are adversely impacted by increased rates of tobacco use. Rural residents may be exposed to unique communal norms and other factors that influence smoking cessation. PURPOSE: This study explored facilitating factors and barriers to cessation and the role of rural health care systems in the smoking-cessation process. METHODS: Focus groups were conducted with smokers (N = 63) in 7 Midwestern rural communities. Qualitative analysis and thematic coding of transcripts was conducted. FINDINGS: Three levels of pertinent themes- intrinsic, health-system resource, and community/social factors--were identified. Intrinsic factors facilitating cessation included willingness to try various cessation methods, beliefs about consequences of continuing smoking (eg, smoking related illnesses), and benefits of quitting (eg, saving money). Intrinsic barriers included skepticism about resources, low self-efficacy and motivation for smoking cessation, concern about negative consequences of quitting (eg, weight gain), and perceived benefits of continued smoking (eg, enjoyment). Key health-system resource facilitators were pharmacotherapy use and physician visits. Resource barriers included infrequent physician visits, lack of medical/financial resources, limited local smoking-cessation programs, and lack of knowledge of existing resources. In terms of community/social factors, participants acknowledged the negative social impact/image of smoking, but also cited a lack of alternative activities, few public restrictions, stressors, and exposure to other smokers as barriers to cessation. CONCLUSIONS: Smokers in rural communities face significant challenges that must be addressed. A multilevel model centered on improving access to health care system resources while addressing intrinsic and community/social factors might enhance smoking-cessation interventions and programs in rural communities. PMID- 18397445 TI - Kansas primary care weighs in: a pilot randomized trial of a chronic care model program for obesity in 3 rural Kansas primary care practices. AB - CONTEXT: Obesity is a chronic disease of epidemic proportions in the United States. Primary care providers are critical to timely diagnosis and treatment of obesity, and need better tools to deliver effective obesity care. PURPOSE: To conduct a pilot randomized trial of a chronic care model (CCM) program for obesity care in rural Kansas primary care. METHODS: We enrolled 107 participants to a 6-month, 2-armed, randomized trial comparing a CCM for obesity with usual care. The primary outcome was weight change at 90 days. The usual care arm received educational weight loss materials and outcome assessments at day 0, 90, and 180. The active arm received the same elements as the usual care arm plus a multicomponent obesity CCM. FINDINGS: The Day 90 mean +/- SD weight change for the active arm (n = 34) and control arm (n = 33), respectively, was -4.5 +/- 7.7 pounds and -2.4 +/- 8.1 pounds (P = .27 for difference). The Day 180 mean +/- SD weight change for the active (n = 27) and control (n = 27) arms, respectively, was -9.4 +/- 10.3 pounds and -2.1 +/- 10.7 pounds (P = .01 for difference). There was no significant change in physical activity, or fruit and vegetable intake at day 90 or day 180. CONCLUSIONS: Improving the recognition and treatment of obesity in primary care settings is a critical initiative. Rural populations suffer disproportionately with obesity, and better methods of delivering obesity care are needed for this population. Further research is needed to establish the effectiveness of a CCM approach for obesity care. PMID- 18397446 TI - Vital signs screening for alcohol misuse in a rural primary care clinic: a feasibility study. AB - CONTEXT: Alcohol misuse is more common in rural areas, and rural problem drinkers are less likely to seek alcohol treatment services. Rural clinics face unique challenges to implementing routine alcohol screening and intervention. PURPOSE: To assess the feasibility of using the single alcohol screening question (SASQ) during routine nursing vital signs in a rural clinic, and to determine its effect on alcohol screening and intervention rates. METHODS: Patient exit interviews were used to identify alcohol misuse and to measure changes in screening and intervention rates. Chi-square tests were used to compare rates of screening across study phases, while odds ratios from logistic regression analyses were used to quantify association between nurse screening and clinician intervention. FINDINGS: Exit interviews were completed by 126 current drinkers (41 before vital signs screening implementation and 85 afterward). Screening rates for alcohol misuse rose from 14.6% at baseline to 20.0% (P = .027) after screening implementation. Clinician intervention rates among alcohol misusers rose from 6.3% to 11.8% (P = .039). Nurse screening increased the odds of clinician intervention (OR 1.47; 95% CI 1.10-1.95). CONCLUSIONS: Vital signs screening proved to be feasible in this rural clinic and produced modest but significant increases in alcohol screening by nurses and brief interventions by clinicians. Additional studies are needed to define effective strategies for further increasing these rates. PMID- 18397447 TI - Physical activity, metabolic syndrome, and overweight in rural youth. AB - BACKGROUND: Research suggests significant health differences between rural dwelling youth and their urban counterparts with relation to cardiovascular risk factors. This study was conducted to (1) determine relationships between physical activity and markers of metabolic syndrome, and (2) to explore factors relating to physical activity in a diverse sample of rural youth. METHODS: Data were collected from 4th, 6th, 8th, and 11th grade public school students in the rural Southeastern United States in the spring of 2002. Physiological data included anthropometrics, fasting glucose, lipids, hemodynamics, and skinfold measurements. Psychosocial data included parental support for physical activity, accessibility of physical activity facilities, and safety concerns for physical activity. Behavioral data included self-reported physical activity and sedentary behaviors. RESULTS: After adjusting for sex, race, and age, subjects with low level of physical activity were 3 times more likely to be positive for metabolic syndrome compared to those reporting a high level of physical activity. Subjects reporting a low level of physical activity were 2.4 times more likely to be overweight compared to subjects reporting a high level of physical activity. Students with high levels of physical activity were more likely to have parents who provided money for physical activity lessons and sports teams. CONCLUSIONS: Rural youth with low levels of physical activity participation were at increased risks for metabolic syndrome and overweight. Effective physical activity promotions addressing supports for physical activity are urgently needed in rural America. PMID- 18397448 TI - Alcohol consumption and injury among Canadian adolescents: variations by urban rural geographic status. AB - CONTEXT: The impact of alcohol consumption on risks for injury among rural adolescents is an important and understudied public health issue. Little is known about whether relationships between alcohol consumption and injury vary between rural and urban adolescents. PURPOSE: To examine associations between alcohol and medically attended injuries by urban-rural geographic status using a representative national sample of Canadian adolescents. METHODS: The study involved a secondary analysis of a national sample of Canadian adolescents aged 11-15 years (n = 7,031) from the 2001-2002 Health Behavior in School-Aged Children Survey. Respondents were classified into 5 geographic categories of rural-urban status. Multiple logistic regression was used to examine the magnitude and homogeneity of associations between drinking patterns and adolescent injuries across these 5 geographic groupings. FINDINGS: Higher rates of alcohol consumption and adolescent injuries were observed in more rural areas. Alcohol consumption was significantly associated with higher risks for injury occurrence with evidence of a dose-related pattern of risk. Associations between alcohol consumption and injury were consistent by urban-rural geographic status. CONCLUSIONS: Misuse of alcohol is an important potential cause of injury. Adolescents whose lifestyle includes alcohol consumption experience higher risks for injury, and this association is observed consistently by urban-rural geographic status. Findings of this study emphasize a need to intervene with high risk adolescents as a tertiary prevention strategy, irrespective of geographic background. PMID- 18397449 TI - The occupational mix adjustment to the Medicare hospital wage index: why the rural impact is less than expected. AB - CONTEXT: Rural hospitals are heavily dependent on Medicare for their long-term financial solvency. A recent change to Medicare prospective payment system reimbursement--the occupational mix adjustment (OMA) to the wage index--has attracted a great deal of attention in rural policy circles. PURPOSE: This paper explores variation in the OMA across and within urban and rural markets. Reasons why the effect of the OMA has been less than some rural advocates anticipated are discussed. METHODS: Data were obtained from the fiscal year 2007 Final Occupational Mix Survey Data Public Use File and the fiscal year 2007 Final Rule Wage Data Public Use File. Descriptive statistics were generated to determine the need for the OMA and the potential impact of its application on hospitals located in rural markets. FINDINGS: The average OMA for nonmetropolitan markets is greater than 1, indicating that hospitals in these markets use a less-skilled mix of labor than the national average. However, almost one third of nonmetropolitan markets had an OMA that was less than 1 and experienced a net decrease in Medicare reimbursement due to the OMA. CONCLUSIONS: There are several reasons why the impact of the OMA is smaller than many rural hospital administrators expected. The most important is that the adjustment happens at the market-level rather than for individual hospitals, so a small hospital's staffing mix may have almost no effect on the final payment adjustment. In rural markets, it appears that hospitals in micropolitan areas exert a large influence on the OMA. PMID- 18397450 TI - Medicare hospital charges in the last year of life: distribution by quarter for rural and urban nursing home decedents with cognitive impairment. AB - BACKGROUND: Medicare beneficiaries incur 27%-30% of lifetime charges in the last year of life; most charges occur in the last quarter. Factors associated with high end-of-life Medicare charges include less advanced age, non-white race, absence of advance directive, and urban residence. METHODS: We analyzed Medicare hospital charges in the last year of life for nursing home residents with severe cognitive impairment, focusing on rural-urban differences. The study population consisted of 3,703 nursing home residents (1,882 rural, 1,821 urban) in Minnesota and Texas who died in 2000-2001. Data on Medicare hospital charges were obtained from 1998-2001 Centers for Medicare and Medicaid Services MedPAR files. RESULTS: During the last year of life, unadjusted charges averaged $12,448 for rural subjects; $31,780 for urban. The charges were distributed across the last 4 quarters similarly for the 2 populations, with 15%-20% of charges incurred in each of the first 3 quarters, and 47% (rural) and 52% (urban) in the last quarter. At the individual level, a higher percentage of hospital charges were incurred in the last 90 days by urban than by rural residents (P < .001). A larger proportion of urban (43%) than rural (37%) residents were hospitalized in the final quarter. The charges for hospitalized residents (N = 1,994) were distributed similarly to those of the entire study population. DISCUSSION: Medicare hospital charges during the last year of life were lower for rural nursing home residents with cognitive impairment than for their urban counterparts. Charges tend to be more concentrated in the last 90 days of life for urban residents. PMID- 18397451 TI - Rural-urban differences in primary care physicians' practice patterns, characteristics, and incomes. AB - CONTEXT: Low salaries and difficult work conditions are perceived as a major barrier to the recruitment of primary care physicians to rural settings. PURPOSE: To examine rural-urban differences in physician work effort, physician characteristics, and practice characteristics, and to determine whether, after adjusting for any observed differences, rural primary care physicians' incomes were lower than those of urban primary care physicians. METHODS: Using survey data from actively practicing office-based general practitioners (1,157), family physicians (1,378), general internists (2,811), or pediatricians (1,752) who responded to the American Medical Association's annual survey of physicians between 1992 and 2002, we used linear regression modeling to determine the association between practicing in a rural (nonmetropolitan) or urban (standard metropolitan statistical area) setting and physicians' annual incomes after controlling for specialty, work effort, provider characteristics, and practice characteristics. FINDINGS: Rural primary care physicians' unadjusted annual incomes were similar to their urban counterparts, but they tended to work longer hours, complete more patient visits, and have a much greater proportion of Medicaid patients. After adjusting for work effort, physician characteristics, and practice characteristics, primary care physicians who practiced in rural settings made $9,585 (5%) less than their urban counterparts (95% confidence intervals: -$14,569, -$4,602, P < .001). In particular, rural practicing general internists and pediatricians experienced lower incomes than did their urban counterparts. CONCLUSIONS: Addressing rural physicians' lower incomes, longer work hours, and greater dependence on Medicaid reimbursement may improve the ability to ensure that an adequate supply of primary care physicians practice in rural settings. PMID- 18397452 TI - Expanding the role of nurse practitioners: effects on rural access to care for injured workers. AB - CONTEXT: A 3-year pilot program to expand the role of nurse practitioners (NPs) in the Washington State workers' compensation system was implemented in 2004 (SHB 1691), amid concern about disparities in access to health care for injured workers in rural areas. SHB 1691 authorized NPs to independently perform most functions of an attending physician. PURPOSE: The aims of this study were to (1) describe the contribution by NPs to Washington's workers' compensation provider workforce, (2) evaluate change in provider availability attributable to SHB 1691, and (3) evaluate the effect of SHB 1691 on timely accident report filing. METHODS: Administrative data were used to evaluate this natural experiment, using a pre-post design with primary care physicians (PCPs) as a nonequivalent comparison group. FINDINGS: NPs served injured workers with characteristics similar to those served by PCPs, but 22.0% of NPs were rural, compared with 17.3% of PCPs. Of claimants with NPs as their attending provider, 53.3% were injured in a rural county, compared with 24.7% for those with PCP attending providers. The number of NPs participating in the workers' compensation system rose after SHB 1691 implementation, more so in rural areas. SHB 1691 implementation was associated with a 16 percentage point improvement in timely accident report filing by NPs in both rural and urban areas. CONCLUSIONS: Authorizing NPs to function as attending providers for injured workers may improve provider availability (especially in rural areas) and timely accident report filing, which in turn may improve worker outcomes and system costs. PMID- 18397454 TI - Nonemergency medicine-trained physician coverage in rural emergency departments. AB - CONTEXT: Rural areas have fewer physicians compared to urban areas, and rural emergency departments often rely on community or contracted providers for staffing. The emergency department workforce is composed of a variety of physician specialties and clinicians. PURPOSE: To determine the distribution of emergency department clinicians and the proportion of care they provide across the rural-urban continuum. METHODS: Cross-sectional analysis of secondary data. The distribution of clinicians who provide emergency department care by county was determined using the 2003 Area Resource File. The percentage of emergency department care provided by clinician type was determined using 2003 Medicare claims data. Logistic regression analyses assessed the odds of being seen by different clinicians with a patient's rurality when presenting to the emergency department. FINDINGS: Board-certified emergency physicians provide 75% of all emergency department care, but only 48% for Medicare beneficiaries of the most rural of counties. The bulk of the remainder of emergency department care is largely provided by family physicians and general internists, with the percentage increasing with rurality. The likelihood of being seen by an emergency physician in the emergency department decreases 5-fold as rurality increases, while being seen by a family physician increases 7-fold. CONCLUSION: Nonemergency physicians provide a significant portion of emergency department care, particularly in rural areas. Medical specialties must cooperate to ensure the availability of high quality emergency department care to all Americans regardless of physician specialty. PMID- 18397453 TI - Mental health professional shortage areas in rural Appalachia. AB - CONTEXT: Research on health disparities in Appalachia has rarely compared Appalachia to other geographic areas in such a way as to isolate possible Appalachian effects. PURPOSE: This study tests hypotheses that nonmetropolitan Appalachia will have higher levels of mental health professional shortage areas than other nonmetropolitan areas of the same states, but that these disparities will dissipate when accounting for social and economic differences. METHODS: The study analyzed secondary data for nonmetropolitan counties (N = 618) in the 13 Appalachian states. Appalachian counties were identified from the Appalachian Regional Commission designations. Mental health professional shortages were identified from Health Resources and Services Administration data. Area Resource File data were used to measure county-level income, education, uninsurance, unemployment, race/ethnicity percentages, and urban influence codes. Logistic regression models tested whether Appalachia was significantly associated with shortage areas, and whether the Appalachian effect persisted after accounting for social and economic covariates. FINDINGS: Seventy percent of Appalachian nonmetropolitan counties were mental health professional shortage areas, significantly higher than non-Appalachian, nonmetropolitan counties in the same states. The Appalachian effect did not persist after controlling for the full set of other variables; education and race/ethnicity emerged as significant predictors. CONCLUSIONS: Appalachia location is associated with mental health professional shortages, but this effect is driven by underlying social differences, in particular by lower education. This method of identifying Appalachia for comparative purposes may be applied to many other health services research questions and to other defined geographic regions. PMID- 18397455 TI - Stroke knowledge among urban and frontier first responders and emergency medical technicians in Montana. AB - PURPOSE: To assess stroke knowledge and practice among frontier and urban emergency medical services (EMS) providers and to evaluate the need for additional prehospital stroke training opportunities in Montana. METHODS: In 2006, a telephone survey of a representative sample of EMS providers was conducted in Montana. Respondents were stratified into 2 groups: those working in urban and frontier counties. FINDINGS: Compared to EMS providers from urban counties, those from frontier counties were significantly more likely to be older (mean age 44.7 vs 40.1 years), have fewer personnel working in their service (mean 17.7 vs 28.6), to be located farther away from a computed tomography scan (CT scan) (mean 41.3 vs 17.6 miles), and to be volunteers (84% vs 49%). They were also less likely to have a stroke protocol (58% vs 66%) and use a stroke screening tool (36% vs 47%) than their urban counterparts. There were no significant differences between frontier and urban EMS respondents' ability to correctly identify 4 or more stroke warning signs (58% vs 61%), 4 or more stroke risk factors (46% vs 43%), or the 3-hour recombinant tissue plasminogen activator (rt-PA) treatment window (56% vs 57%). Approximately two thirds of respondents from urban and frontier counties believed they had adequate stroke knowledge, but 90% indicated they were interested in additional stroke-related training. CONCLUSIONS: Although stroke knowledge did not differ between urban and frontier groups, stroke screens and stroke protocols were less likely to be used in the frontier areas. Training opportunities and the implementation of stroke protocols and screening tools are needed for EMS providers, particularly in frontier counties. PMID- 18397456 TI - Uninsured hospitalizations: rural and urban differences. AB - CONTEXT: Few studies have examined hospitalization patterns among the uninsured, especially from the perspective of rural and urban differences. PURPOSE: To examine whether the patterns of uninsured hospitalizations differ in rural and urban hospitals and to identify the most prevalent and costly diagnoses among uninsured hospitalizations. METHODS: We conducted a cross-sectional analysis of the Healthcare Cost and Utilization Project's National Inpatient Sample representing a total of 37,804,021 hospital discharges, with 4.9% of them generated by uninsured persons in 2002. We compared demographic and clinical characteristics and the proportion of frequent and costly diagnoses by rural and urban hospitals. We used multiple logistic regression models to examine the relationship between preventable conditions and rural and urban hospitals among uninsured hospitalizations. FINDINGS: Uninsured persons discharged from rural hospitals were more likely than their urban counterparts to be working-age adults (82% vs 79%) and to reside in a ZIP code area with a median household income of less than $35,000 per year (56% vs 26%). Rural uninsured hospitalizations were more likely to be for preventable conditions than were urban uninsured hospitalizations (P < .001). The proportion of total hospital charges related to preventable hospitalizations was 15.5% in rural hospitals versus 10.0% in urban hospitals. CONCLUSIONS: The patterns of uninsured hospitalizations in rural and urban hospitals were different in many ways. Providing adequate access to primary care could result in potential savings related to preventable hospitalizations for the uninsured, especially for rural hospitals. PMID- 18397457 TI - Rural residence and prescription medication use by community-dwelling older adults: a review of the literature. AB - CONTEXT: Due to various barriers to health care access in the rural setting, there is concern that rural older adults might have lower access to prescribed medications than their urban counterparts. PURPOSE: To review published research reports to determine prevalence and mean medication use in rural, noninstitutionalized older adults and assess whether rural-urban differences exist. METHODS: PubMed, Ageline, Cinahl, PsycInfo, International Pharmaceutical Abstracts, Agricola, and Institute for Scientific Information Web of Science - Social Science Index were searched. English-language articles through May 2005 involving a sample of rural, noninstitutionalized older adults and analyses of overall medication prevalence and/or intensity were included. Review articles, conference abstracts, dissertations, books, and articles targeting nonprescription or specific therapeutic categories were excluded. A total of 206 citations were identified and 26 met the inclusion criteria. FINDINGS: Reported prevalence of prescription medication use by rural older adults varied between 62% and 96%, with 2-6 prescriptions per person. Multivariate analyses results were equally inconsistent. Controlling for insurance, most US studies suggest there is no rural-urban difference in access to prescribed medications. However, this finding may not be generalizable across all regions in the United States or other countries. CONCLUSIONS: Geographic location may not be as important a variable for medication usage as for other health services utilization. PMID- 18397458 TI - Determinants of rural women's health: a qualitative study in Southwest Ontario. AB - CONTEXT: The influences of gender and geography are increasingly being acknowledged as central to a comprehensive understanding of health. Since little research on rural women's health has been conducted, an in-depth qualitative approach is necessary to gain a better initial understanding of this population. PURPOSE: To explore the determinants of health and their influence on rural women's health. METHODS: From November 2004 to September 2005, 9 focus groups and 3 individual interviews were conducted in 7 rural southwestern Ontario communities. Sixty-five rural residents aged 26 years and older participated in the study. Semi-structured interview questions were used to elicit participants' perceptions regarding determinants of rural women's health. FINDINGS: Four Health Canada determinants (employment, gender, health services, and social environments) and 3 new determinants (rural change, rural culture, and rural pride) emerged as key to rural women's health. CONCLUSIONS: Although health determinants affect both urban and rural people, this qualitative study revealed that rural women experience health determinants in unique ways and that rural residents may indeed have determinants of their health that are particular to them. More research is needed to explore the nature and effects of determinants of health for rural residents in general, and rural women in particular. PMID- 18397459 TI - Reason for visit: is migrant health care that different? AB - PURPOSE: The purpose of this pilot study was to determine the reasons for which migrant agricultural workers in Pennsylvania seek health care. METHODS: Participants were individuals 14 years of age and over, actively involved in agricultural labor and presenting for medical care at 6 migrant health care centers. Bilingual health care providers randomly selected and interviewed the participants. FINDINGS: The most commonly reported reason for visiting the health care provider was for physical examination. The most frequent acute problems were related to the musculoskeletal and integumentary systems. Frequently cited problems in the medical history were hypertension, musculoskeletal/back pain, and gastrointestinal conditions. Most medications being taken were for cardiovascular or pain-related problems. CONCLUSIONS: These results suggest that migrant workers present with medical problems that are similar to those of the general primary care population. Many problems were recurrent and represented common chronic medical conditions. PMID- 18397460 TI - Disturbed expression of E-cadherin, beta-catenin and tight junction proteins in colon carcinoma is unrelated to growth pattern and genetic polymorphisms. AB - Adhesion proteins are responsible for the structural integrity of epithelial tissue and in tumors this integrity is often lost, resulting in a disorganization of the tissue. In the present study the complexity of the invasive front of colon carcinomas was correlated with cell adhesion protein expression and with polymorphisms in their genes. A complexity index was constructed from 32 colon carcinomas using computer-assisted morphometry estimating fractal dimension and tumor cell clusters followed by tree analysis. Immunohistochemical staining of beta-catenin, E-cadherin, occludin and claudin 2 was used for assessment of protein expression. Genetic screening of tissue from the tumor invasion front with laser microdissection was performed using SSCP and DNA sequencing. Adhesion protein distribution was significantly disturbed in most carcinomas. A single mutation in the gene of beta-catenin was found but there was no correlation between protein expression and genetic polymorphism. Nor was there any correlation between the complexity of the invasive border and protein distribution or genetic alterations. The results indicate that the complexity of colon carcinoma invasion is not dependent on genetic derangements in the genes of adhesion proteins or the protein distribution. Rather, aberrations in the function of other proteins related to the adhesive proteins could be responsible. PMID- 18397461 TI - The lower genital tract microbiota in relation to cytokine-, SLPI- and endotoxin levels: application of checkerboard DNA-DNA hybridization (CDH). AB - In the present study the lower genital tract microbiota in asymptomatic fertile women (n=34) was identified and quantified by culturing vaginal secretions. Also, vaginal and cervical samples were analyzed by a semiquantitative checkerboard DNA DNA hybridization technique (CDH) based on genomic probes prepared from 13 bacterial species (Bacteroides ureolyticus, Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Mobiluncus curtisii ss curtisii, Prevotella bivia, Prevotella disiens, Prevotella melaninogenica, Atopobium vaginae, Lactobacillus iners, Staphylococcus aureus ss aureus, Streptococcus anginosus, and Streptococcus agalactiae). The bacterial species found by either culture or CDH were correlated with proinflammatory cytokines (IL-1 alpha, IL-1 beta, IL-6, IL-8), secretory leukocyte protease inhibitor (SLPI), and endotoxin in the cervicovaginal samples. Grading the women into healthy, intermediate, or bacterial vaginosis (BV) as based on Gram staining of vaginal smears, the viable counts of lactobacilli (L. gasseri) and of streptococci-staphylococci combined were highest in the intermediate group. In BV, particularly the high concentrations of Actinomyces urogenitalis, Atopobium vaginae, and Peptoniphilus harei were noted (>or=10(11) per ml). The total viable counts correlated with both cervical IL-1 alpha and IL-1 beta. A strong negative correlation was observed between L. iners and total viable counts, G. vaginalis, or cervical IL-1 alpha, while it correlated positively with SLPI. Analysis of vaginal and cervical samples from 26 out of the 34 women by CDH showed that anaerobic bacteria were more frequently detected by CDH compared to culture. By this method, A. vaginae correlated with G. vaginalis, and L. iners with S. aureus. With regard to cytokines, B. ureolyticus correlated with both cervical and vaginal IL-1 alpha as well as with cervical IL-8, while F. nucleatum, S. agalactiae, S. anginosus, or S. aureus correlated with vaginal IL-1 alpha. Furthermore, all Gram-negative bacteria taken together, as measured by CDH, correlated with vaginal endotoxin and inversely with vaginal SLPI. The significance of the results is discussed. In summary, mapping of the identity and quantity of vaginal bacterial species and their association with locally produced host innate immune factors will help in defining various types of abnormal vaginal microbiota, developing new ways of assessing the risk of ascending subclinical infections, and in treating them. CDH appears to be a suitable tool for future analyses of large numbers of clinical samples with an extended number of bacterial probes. PMID- 18397462 TI - Risk of miscarriage for pregnant users of pivmecillinam: a population-based case control study. AB - Few data exist on the risk of miscarriage after exposure to pivmecillinam. We therefore conducted a population-based case-control study in a Danish county with 0.5 million inhabitants during the period 1997-2002. We included 1,599 women with a miscarriage recorded in the Hospital Discharge Registry and selected 10 controls per case among primiparous women who had a live birth during the study period. Controls were selected from the Danish Medical Birth Registry. We obtained data on use of pivmecillinam and sulfamethizole from a prescription database. Five cases (0.30%) and 24 controls (0.15%) were exposed to pivmecillinam in the last week before the miscarriage/index date. After adjustment for maternal age, use of antidiabetics or antiepileptics, the odds ratio for miscarriages among users of pivmecillinam compared with non-users was 2.03 (95% confidence interval: 0.77-5.33) and the corresponding odds ratio for use of sulfamethizole was 1.53 (95% confidence interval: 0.76-3.09). Exposure within 2 to 12 weeks before the miscarriage was not associated with an increased risk. We concluded that use of pivmecillinam was associated with an increased risk of miscarriage, but the risk was not significantly (p=0.64) different from the risk associated with use of sulfamethizole. PMID- 18397463 TI - Genotypic profiles and virulence attributes of Candida albicans isolates from patients with oral lichen planus. AB - Candida albicans carriage has been found to be increased in patients with oral lichen planus. In the present work we have investigated the genotypic profiles of 112 C. albicans strains isolated from patients with erosive or nonerosive OLP, and from healthy controls. The virulence attributes of the isolated strains were compared to elucidate the pathogenetic mechanisms through which C. albicans may cause OLP. We have characterized the genotypic profiles of these isolated strains using a randomly amplified polymorphic DNA assay. In addition, we used assays to measure adhesion to buccal epithelial cells and phospholipase activity to evaluate the virulence attributes of these isolates. Our RAPD analyses revealed four different genotypes, named type A to D, among all isolates, and identified statistically significant associations with disease conditions. Specifically, type A (58.1%) and C (29.0%) were primarily found in erosive OLP, while type A (33.3%) and D (58.3%) were identified in nonerosive OLP. However, the healthy group seemed to have type B (38.5%) and D (61.5%). Using adhesion to BEC assay, we demonstrated that the isolates with genotype A had the strongest adherence among the four genotypes (P=0.000<0.05). The phospholipase activity of the isolates with genotype A and C was higher than for those with genotype B and D (P=0.000<0.05). In conclusion, some C. albicans isolates with special genotypic profiles and virulence attributes may contribute to the pathogenesis of OLP. PMID- 18397464 TI - Viral reactivation is not related to septic complications after major surgical resections. AB - Anastomotic leakage and septic complications are the most important determinants of postoperative outcome after major surgical resections. Malignant diseases and surgical trauma can influence immune responses and the ability to react against infectious factors, such as bacteria and viruses. Comparable immune suppression can cause viral reactivation in transplantation and trauma patients. In this prospective study, patients who underwent major surgical resections for oesophageal or pancreatic cancer were investigated for the potential involvement of viral reactivation in the development of septic complications. 86 patients (40 oesophageal resections, 27 pancreatic resections, 19 surgical explorations) were included. Viral antigens, viral DNA, antibodies against viral structures (IgG, IgM, IgA) and, in part, viral cultivation were performed for CMV, EBV, HSV1, HSV2, HZV6 and VZV in serum, urine, sputum and swabs from buccal mucosa preoperatively and at postoperative days 1, 3 and 5. Test results were compared with the postoperative outcome (30-day morbidity, in-hospital mortality) and clinical scores (SOFA, TISS). For statistical analyses Student's t-tests and Chi2 tests were used. The overall complication rate was 19.8% (30-day morbidity) with an in-hospital mortality of 1.2% (1/86 patients). Postoperatively, anti-CMV-IgG titres were significantly reduced (p<0.05) and remained suppressed in patients with septic complications. Anti-CMV-gB-IgG were also reduced, but showed considerable interindividual differences. Anti-CMV-IgA and -IgM did not show significant alterations in the postoperative course. In addition, direct viral detection methods did not support viral reactivation in patients in any of the investigated groups. The reduction of anti-CMV antibodies is likely caused by an immune suppression, specifically by reduced B-cell counts after major surgical interventions. Viral reactivation, however, did not occur in the early postoperative period as a specific risk for septic complications. PMID- 18397465 TI - The first major extended-spectrum beta-lactamase outbreak in Scandinavia was caused by clonal spread of a multiresistant Klebsiella pneumoniae producing CTX-M 15. AB - Between May and December 2005, 64 multidrug-resistant isolates of Klebsiella pneumoniae were detected from patients admitted to Uppsala University Hospital. This represented a dramatic increase in ESBL-producing K. pneumoniae compared to previous years. To investigate the epidemiology and to characterize the resistance mechanisms of the isolates, a study was initiated. Antibiotic susceptibility was determined by means of the Etest and the disc diffusion method. Extended-spectrum beta-lactamase (ESBL) production was identified by clavulanic acid synergy test and confirmed with PCR amplification followed by DNA sequencing. DNA profiles of the isolates were examined with pulsed-field gel electrophoresis (PFGE). All isolates were resistant or exhibited reduced susceptibility to cefadroxil, cefuroxime, cefotaxime, ceftazidime, aztreonam, piperacillin/tazobactam, ciprofloxacin, tobramycin, and trimethoprim sulfamethoxazole. They produced ESBL of the CTX-M-15 type, and the involvement of a single K. pneumoniae clone was shown. This is the first major clonal outbreak of multiresistant ESBL-producing K. pneumoniae in Scandinavia. The outbreak demonstrates the epidemic potential of enterobacteria containing ESBLs of the CTX M type, even in a country with a relatively low selective pressure and a low prevalence of multiresistant bacteria. PMID- 18397466 TI - Immunoexpression and prognostic role of hTERT and cyclin D1 in urothelial carcinoma. AB - The aim was to investigate the expression of human telomerase reverse transcriptase (hTERT) and cyclin D1 in correlation with clinicopathologic features of urothelial carcinoma (UC). Tissue microarrays (TMA) were constructed from paraffin-embedded specimens of 94 UC patients and immunohistochemical staining was used. High hTERT expression was found in 50 (53%) of the 94 tumors and was significantly associated with tumor invasiveness and tumor grade (P=0.008 and 0.0190, respectively). High cyclin D1 expression was found in 69 (73%) of the 94 tumors and was significantly associated with non-invasiveness and smaller tumor size, but there was no correlation with tumor grade. Kaplan-Meier analysis indicated that patients with low hTERT and high cyclin D1 levels had longer local recurrence-free survival (P=0.0482 and 0.0123, respectively). In addition, patients with high cyclin D1 levels had longer disease-free survival (P=0.0195). In conclusion, this study demonstrated that hTERT and cyclin D1 may be of recurrence predictive value for UC, thus providing clinicians with ancillary information when deciding on suitable therapeutic strategies in UC. PMID- 18397467 TI - Genotyping of respiratory syncytial virus (RSV) group A in Stockholm, Sweden, using PCR and two-dimensional melting curve analysis. AB - Genotyping of respiratory syncytial (RS) virus group A, by means of a novel method based on PCR, FRET (fluorescence resonance energy transmission) detection and two-dimensional melting curve analysis, was carried out on 80 RS virus samples of group A collected in Stockholm from 1976 to 2005. The Tm values were assessed for three different genotypes (GA2, GA5 and GA7) circulating in Sweden. Two pairs of probes were used and results of subsequent data analysis were plotted in a two-dimensional system. The results obtained were compared to genotyping using conventional nucleotide sequencing and phylogenetic tree analysis. It was found that the new assay was able to correctly identify genotype in about 89% of the isolates; it identified the remaining 11% as untypeable and as candidates for conventional nucleotide sequencing. The new method constitutes a complement to nucleotide sequencing and could be useful in studies of large numbers of samples in epidemiological studies. PMID- 18397468 TI - Giant chondroid hamartoma in a 19-year-old girl. PMID- 18397469 TI - Signet ring cell carcinoma of the eyelid - the monocle tumour. AB - We report the clinical and histopathological characteristics of two cases of signet ring cell carcinoma of the eye lids, and discuss the histogenesis of this neoplasm. Two 72-year-old Caucasian males both presented with slowly growing tumours of the eyelids. The tumours were excised and specimens were examined using light- and transmission electron microscopic techniques. Clinically, the tumours infiltrated both eyelids on one side of the face with swelling and periocular inflammation, creating a monocle-like appearance. Extensive clinical work-up excluded periocular metastases. Histopathologically, the tumours were composed of rather bland cells with mainly histiocytoid morphology. A minor proportion had a signet ring cell appearance. The cytoplasmic inclusions giving the signet ring morphology were PAS- and colloidal iron positive. The tumour cells reacted with antibodies against cytokeratins, carcinoembryonic antigen, epithelial membrane antigen, gross cystic disease fluid protein-15 and lysozyme. Transmission electron microscopy demonstrated tumour cells containing intracytoplasmic vacuoles lined by microvilli. The tumour cells aggregated in duct-like clusters. A diagnosis of primary signet ring cell carcinoma was made in both cases. Histopathological, immunohistological and ultrastructural findings indicated that the tumours were of sweat gland origin. PMID- 18397470 TI - Mutational analysis of the BRAF gene in human tumor cells. AB - Genes of the RAF family, which mediate cellular responses to growth signals, encode kinases that are regulated by RAS and participate in the RAS, RAF, mitogen/extracellular signal-regulated kinase, extracellular signal-regulated kinase and mitogen-activated protein kinase pathway. As BRAF is a serine/threonine kinase that is commonly activated by somatic point mutation, it may provide possible diagnostic and therapeutic targets in human malignant tumors. We analyzed exon 15 of the BRAF gene for mutations in 58 lung, 12 breast, six kidney, 14 cervical, four endometrial and 10 ovarian carcinoma cell lines by PCR-SSCP and direct sequencing. The T1796A transversion was found in one (2.9%) of 34 small cell lung carcinoma and one (8.3%) of 12 breast carcinoma cell lines, resulting in a valine-to-glutamate substitution at residue 599 (V599E). One (4.2%) of 24 non-small cell lung carcinoma cell line showed the C1786G transversion, leading to a leucine-to-valine substitution at residue 596 (L596V). No BRAF point mutations were found in any of the other cell lines examined. Our present results suggest that BRAF may not be a frequent target of mutations involved in the pathogenesis of human lung, breast, kidney, cervical, endometrial and ovarian carcinomas. PMID- 18397471 TI - Alteration and preservation of cellular characteristics in long-term culture of tetraploid H-1 (ES) cells. AB - To examine the alteration in cellular characteristics of polyploid ES cells during long-term culturing, tetraploid H-1 (ES) cells were continuously cultured for 180 days. Cellular DNA content of the tetraploid cells decreased and reached a plateau of 3.3 C, where C represents the complement of haploid chromosomes. The chromosome number also decreased, indicating that the DNA loss was induced by chromosome loss. Cell volume was maintained, suggesting that the DNA loss did not involve cytoplasmic loss. The cell cycle parameters were almost the same during the DNA decay process, indicating that cell cycle progression was independent of the quantity of homologous chromosomes. Hypotetraploid cells showed alkaline phosphatase activity and formed teratocarcinomas in mouse abdomens, suggesting that the pluripotent potential was maintained. Cellular morphology was also retained, suggesting that the gene expression specifying morphological characteristics was conserved. We conclude that these initial cellular characteristics of tetraploid H1 (ES) cells were preserved in long-term culture, irrespective of chromosome loss. PMID- 18397472 TI - G1 arrest and expression of cyclin-dependent kinase inhibitors in tamoxifen treated MCF-7 human breast cancer cells. AB - Treatment of exponentially growing MCF-7 human breast carcinoma cells with tamoxifen (TAM) inhibits cell growth in a dose-dependent manner. However, the molecular basis for the drug's activity and its relationship to the cell cycle have not yet been clearly established. In this study, we analyzed cell cycle related proteins used for immunoblotting and flow cytometry in TAM-treated MCF-7 cells. In addition, the ratio of apoptosis in the cell was analyzed using labeling of DNA strand breaks (TdT assay). In flow-cytometric DNA distribution analysis, the S-phase fraction showed a marked decrease and a concomitant increase in G1- and G2-phase cells accompanying the inhibitory effect of TAM; these changes were time- and dose-dependent. Immunoblotting revealed that the levels of p53 and p21(WAF1/CIP1) in TAM-treated cells increased in a time- and dose-dependent manner, whereas those of p27(KIP1) and p16 slightly increased or remained unchanged. Furthermore, cyclin D3 and B showed sharp decreases, in contrast with p53 and p21(WAF1/CIP1) DNA-apoptosis dual analysis using flow cytometry revealed that the TAM-treated samples contained apoptotic cells, the majority of which were arrested in G1 or G2 and showed suppression of Bcl-2 protein. These results suggest that the tumorigenic effect of TAM on MCF-7 cells arises through antitumor effects that are due to the expression of cyclin dependent kinase inhibitors, especially p21(WAF1/CIP1) and these are regulated by the decrease of wild-type p53. The proposed mechanism is similar to that underlying the cytotoxic effects of other agents and ionizing irradiation that cause DNA damage. PMID- 18397473 TI - Induced in-vitro differentiation of neural-like cells from human amnion-derived fibroblast-like cells. AB - There is growing evidence that the human amnion contains various types of stem cell. As amniotic tissue is readily available, it has the potential to be an important source of material for regenerative medicine. In this study, we evaluated the potential of human amnion-derived fibroblast-like (HADFIL) cells to differentiate into neural cells. Two HADFIL cell populations, derived from two different neonates, were analyzed. The expression of neural cell-specific genes was examined before and after in vitro induction of cellular differentiation. We found that neuron specific enolase, neurofilament-medium, beta-tubulin isotype III, and glial fibrillary acidic protein (GFAP) showed significantly increased expression following the induction of differentiation. In addition, immunostaining demonstrated that neuron specific enolase, GFAP and myelin basic protein (MBP) were present in HADFIL cells following the induction of differentiation, although one of the HADFIL cell populations showed a lower expression of GFAP and MBP. These results indicate that HADFIL cell populations have the potential to differentiate into neural cells. Although further studies are necessary to determine whether such in vitro-differentiated cells can function in vivo as neural cells, these amniotic cell populations might be of value in therapeutic applications that require human neural cells. PMID- 18397474 TI - Establishment and characterization of a cell line (OMC-9) originating from a human endometrial stromal sarcoma. AB - Cell lines are very useful for clinical and basic research. The establishment of uterine malignant tumor cell lines with unusual histology is especially important. We describe the establishment and characterization of a new human endometrial stromal sarcoma cell line of the uterus. The cell line OMC-9 was established from a tumor mass in the uterine body of a 55-year-old woman. Characteristics of the cell line studied include morphology, chromosome analysis, heterotransplantation, tumor markers and chemosensitivity. This cell line has grown well for 196 months and has been subcultured more than 50 times. Monolayer cultured cells are polygonal in shape, appear to be spindle-shaped or multipolar and have a tendency to pile up without contact inhibition. The cells exhibit a human karyotype with a modal chromosomal number in the diploid range. The cells were able to be transplanted into the subcutis of nude mice and produced tumors resembling the original tumor. OMC-9 cells produced tissue polypeptide antigen. Both CD10, a sensitive and diagnostically useful marker of endometrial stromal neoplasms, and vimentin were identified immunohistochemically in the original tumor and the heterotransplanted tumor. The cells were sensitive to actinomycin D, doxorubicin, carboplatin, cisplatin and etoposide, drugs used commonly in the treatment of gynecologic cancer. Only three reports of uterine endometrial stromal sarcoma cell lines have thus far been reported in the literature. OMC-9 is the first endometrial stromal sarcoma cell line in which CD10 expression and chemosensitivity have been identified. PMID- 18397478 TI - An important consensus statement on the management of gastroesophageal reflux disease for the largest population in the world. PMID- 18397479 TI - Loss of interstitial cells of Cajal may be central to poor intestinal motility in diabetes mellitus. PMID- 18397480 TI - Pancreatic cancer: tissue is the issue. PMID- 18397481 TI - Primary biliary cirrhosis in Asians: less common than in Europeans, but just as depressing. PMID- 18397482 TI - Hepatitis B virus/hepatitis C virus coinfection: epidemiology, clinical features, viral interactions and treatment. AB - Because of the shared modes of transmission, hepatitis B virus (HBV)/hepatitis C virus (HCV) coinfection is not uncommon in highly endemic areas and among subjects with a high risk of parenteral infections. The worldwide prevalence of HBV/HCV coinfection is unknown and might be underestimated with the phenomenon of silent (occult) HBV infection. HCV superinfection in patients with chronic HBV infection was the most common clinical features of coinfection in Asia-Pacific countries. Further, most, but not all, clinical observations suggested that interference between the two viruses was more frequently characterized by an inhibition of HBV replication exerted by HCV. However, longitudinal follow-up studies have demonstrated that the virological patterns in coinfection cases are widely divergent and have dynamic profiles over time. As compared with monoinfected patients, HBV/HCV coinfected persons tend to have more severe liver injury, a higher probability of liver cirrhosis and hepatic decompensation, and a higher incidence of hepatocellular carcinoma. Detailed serological and virological evaluations are required for coinfected patients before initiation of antiviral therapy. Previous studies demonstrated that HBV/HCV coinfected patients responded poorly to interferon (IFN) monotherapy. Currently, for patients with dominant HCV infection and low level HBV viremia (<10(4) IU/mL), IFN or pegylated IFN plus ribavirin can achieve comparable sustained virus response as expected with HCV monoinfection. However, phenomenon of reciprocal viral interference can happen, and resultant "flare" of hepatitis activity may cause liver function deterioration. For coinfected patients with dually-active HBV/HCV, the optimal regimen for therapy remains unclear although adding oral nucleos(t)ide analogs to pegylated IFN and ribavirin seems a reasonable empiric option. PMID- 18397483 TI - Malnutrition in end stage liver disease: recommendations and nutritional support. AB - Malnutrition has increasingly been acknowledged as an important prognostic factor which can influence the clinical outcome of patients suffering from end-stage liver disease (ESLD). Despite the fact that malnutrition is not included in the Child-Pugh classification, its presence should alert clinicians to the same extent as do other complications, such as ascites and hepatic encephalopathy. The pathophysiological mechanisms and the clinical conditions that drive cirrhotic patients to an ill-balanced metabolic state are multiple and they intertwine. Inadequate offer of nutrients, the hypermetabolic state in cirrhosis, the diminished synthetic capacity of the liver and the impaired absorption of nutrients are the main reasons that disrupt the metabolic balance in ESLD. Identifying patients that are approaching the state of malnutrition by simple and easily applied methods is necessary in order to provide nutritional support to those that need it most. According to the European Society for Clinical Nutrition and Metabolism, simple bedside methods such as Subjective Global Assessment and anthropometric parameters are reliable in assessing the nutritional state of cirrhotic patients. Correcting the nutrient deficit of the affected patients is mandatory. Avoidance of alcohol and excess fat and ingestion of 4-6 meals/day containing carbohydrates and protein are the most common recommendations. In severe malnutrition, initiation of enteral feeding and/or use of special formulae such as branched-chain amino acid-enriched nutrient mixtures are often recommended. Enteral nutrition improves nutritional status and liver function, reduces complications, prolongs survival and is therefore indicated. PMID- 18397484 TI - Prospective study of short-term peginterferon-alpha-2a monotherapy in patients who had a virological response at 2 weeks after initiation of interferon therapy. AB - BACKGROUND AND AIMS: Long-term interferon (IFN) therapy is effective in eliminating hepatitis C virus (HCV). However, it carries the risk of adverse effects and reduced quality of life. To assess whether short-term IFN therapy effectively eliminates HCV, we performed a prospective pilot study of pegylated (peg)IFN-alpha-2a therapy for 8 or 24 weeks. METHODS: After excluding patients with high titers of genotype-1, 55 HCV patients received pegIFN-alpha-2a. Patients who became negative for HCV-RNA at week 2 were allocated to either an 8 week (n = 19) or 24-week (n = 15) course of IFN. We evaluated the efficacy of and tolerance to IFN therapy. RESULTS: The sustained virological response rate was excellent in the two groups (8 weeks, 89.5% [17/19]; 24 weeks, 100% [15/15], respectively,). IFN dose reduction was required in one patient of the 8-week group, but in six patients of the 24-week group (P = 0.028). Treatment was completed by all patients of the 8-week group, but discontinued in five patients of the 24-week group (P = 0.011). CONCLUSIONS: The 8-week IFN therapy is more tolerable than the 24-week therapy and had similar outcomes. Excluding the patients with high titers of genotype-1, we recommend switching to an 8-week course of pegIFN-alpha monotherapy once patients show an ultra rapid virological response at week 2 from the start of IFN therapy. PMID- 18397485 TI - Value of repeat endoscopic ultrasound-guided fine needle aspiration for suspected pancreatic cancer. AB - BACKGROUND AND AIMS: Endoscopic ultrasound-guided fine needle aspiration (EUS FNA) is a safe and accurate technique for diagnosing pancreatic cancer. The value of repeat EUS-FNA in patients with high clinical suspicion for pancreatic cancer after an inconclusive index study is unknown. Our aims were to determine the yield and success of repeat EUS-FNA and the reasons for failure of initial EUS FNA. METHODS: This was a retrospective analysis of prospectively collected data in a tertiary University based referral center for pancreatico-biliary disorders. All patients who underwent more then one EUS-FNA for evaluation of suspected pancreatic cancer over a five and a half year period were included in this analysis. RESULTS: Of the 547 procedures performed on 517 patients, 24 (4.6%) patients underwent 51 repeat EUS-FNA procedures. Initial EUS-FNA was atypical/suspicious in 10 (41.6%), benign in 10 (41.6%), malignant in two (8.3%), and failed/indeterminate in two (8.3%) patients. Eight of 10 (80%) patients with atypical/suspicious findings at initial EUS-FNA were diagnosed with malignancy on repeat EUS-FNA. Of the 10 patients with benign findings at initial EUS-FNA, repeat study diagnosed two (20%) with malignancy and the rest were confirmed benign on long-term follow up (average 530 days, SD 369 days). Of the two patients with indeterminate findings at initial EUS-FNA, repeat study diagnosed one patient with malignant disease and the other with benign disease that was confirmed by long-term follow up. Of the two patients diagnosed with neoplastic disease at initial EUS-FNA, repeat EUS-FNA with immunostains downgraded both to chronic pancreatitis. Repeat EUS-FNA facilitated determination of the true status of disease in 20 of 24 patients (accuracy 84%). Suspected reasons for failed initial EUS-FNA were: coexisting pancreatitis (n = 10; 42%), technical difficulty due to scope positioning in uncinate lesion/sedation failure (n = 4; 16.7%), difficult cytology (partly cystic, extensive necrosis, well-differentiated adenocarcinoma) (n = 4; 16.7%), presence of ascites or collaterals (n = 3; 12.5%), pathologist's interobserver variation (n = 2; 8.33%), and unknown reason in one patient. CONCLUSION: Repeat EUS-FNA is warranted in patients with high clinical suspicion for pancreatic cancer despite indeterminate or negative findings at initial EUS-FNA. PMID- 18397486 TI - Economic evaluation of chronic hepatitis B treatments in Taiwan. AB - BACKGROUND AND AIMS: Chronic hepatitis B (CHB) and its sequelae are major health problems in Taiwan. The purpose of the present study was the economic evaluation of short-duration treatments of CHB and longer duration antiviral treatment for up to 5 years. METHODS: Ten-health state CHB disease progression Markov models were used for hepatitis B e antigen (HBeAg)-positive and HBeAg-negative CHB patients, respectively, that included the emergence of antiviral resistance. The perspective of this economic evaluation was the Taiwan health-care system. Costs and benefits were discounted at 3% per annum. RESULTS: Short-course therapies of up to 1-year treatment had limited impact on improving patient survival. Long term viral suppression with lamivudine and adefovir sequential rescue therapies (including add-on therapies) for up to 5 years were found to be highly cost effective by international standards (estimated to be NT$580,000 per quality adjusted life year [QALY] for Taiwan). When Taiwan-specific model inputs were used for HBeAg-positive CHB, the cost per QALY for lamivudine plus adefovir sequential antiviral therapy increased by approximately 100% over the base-case estimate, but was still well within the estimated NT$580,000 per QALY threshold. CONCLUSIONS: In Taiwan, treatment of CHB patients with lamivudine and adefovir sequential antiviral therapies for up to 5 years results in survival benefits and is highly cost-effective. PMID- 18397487 TI - Insulin resistance and lichen planus in patients with HCV-infectious liver diseases. AB - BACKGROUND AND AIM: Hepatitis C virus (HCV) causes liver diseases and extrahepatic manifestations, and also contributes to insulin resistance and type 2 diabetes mellitus (DM). The aims of the present study were to examine the incidence of extrahepatic manifestations including lichen planus in HCV-infected patients and to evaluate the relationship between lichen planus and insulin resistance. METHODS: Of 9396 patients with liver diseases presenting to the study hospital, 87 patients (mean age 60.0 +/- 11.5 years) with HCV-related liver diseases were identified and examined for the incidence of extrahepatic manifestations. Insulin resistance and the presence of Helicobacter pylori antibodies were also measured. RESULTS: The prevalence of DM was 21.8% (19/87), hypertension was 28.7% (25/87), thyroid dysfunction was 20.7% (18/87), and extrahepatic malignant tumor was 9.2% (8/87). The prevalence of lichen planus at oral, cutaneous, pharyngeal, and/or vulval locations was 19.5% (17/87). Characteristics of 17 patients with lichen planus (group A) were compared with 70 patients without lichen planus (group B). Prevalence of smoking history, presence of hypertension, extrahepatic malignant tumor, and insulin resistance (HOMA-IR) were significantly higher in group A than in group B. Significant differences were not observed for age, sex, body mass index, diagnosis of liver disease, alcohol consumption, presence of DM, thyroid dysfunction, liver function tests, or presence of H. pylori infection between the two groups. CONCLUSIONS: Infection with HCV induces insulin resistance and may cause lichen planus. It is necessary for an HCV-infected patient to be assayed for insulin resistance, and to be checked for different extrahepatic manifestations of this infection, particularly lichen planus. PMID- 18397488 TI - Immunohistochemical expression of cell-cycle proteins in gastric precancerous lesions. AB - BACKGROUND: The early indicator for the subject predisposed to gastric cancer is abnormal proliferation of gastric epithelial cells, such as atrophic gastritis (AG), intestinal metaplasia (IM), and dysplasia, which have been considered as precancerous lesions of gastric cancer. To determine whether p53 protein, cyclins D1, and D3, and p27(kip1) play a role in the carcinogenesis pathway of gastric cancer, we performed an immunohistochemical study of their expression in gastric precancerous lesions. METHODS: A total of 1 45 endoscopic gastric biopsy specimens of AG, IM, and gastric dysplasia were studied. These molecular markers were localized by immunohistochemistry. RESULTS: P53 was expressed in 15% of cases with gastric dysplasia and not in the pre-dysplastic stages of the gastric mucosa. All cases were concerning high-grade dysplasia. Cyclin D1 protein was almost undetectable in the precancerous lesions of gastric cancer. Cyclin D3 protein overexpression was seen in 10% of biopsies with IM, and 50% of biopsies with gastric dysplasia. High expression of p27(kip1) protein was demonstrated in all cases of chronic gastritis. As atrophy, IM, and dysplasia develop, expression of p27(kip1) protein is suppressed. In total, 15% of dysplastic cases showed no expression of p27(kip1) protein. CONCLUSIONS: (i) P53 mutation must be a late event during the development of gastric cancer. (ii) Cyclin D1 protein overexpression may not play a role in the progression from normal to neoplastic gastric mucosa, while overexpression of cyclin D3 is an earlier event during gastric carcinogenesis, and its role must be further evaluated. (iii) Reduced expression of p27(kip1) is a rather early event in gastric tumorigenesis, before dysplastic changes occur. PMID- 18397489 TI - Clinicopathological features of gastric cancer detected by endoscopy as part of annual health checkup. AB - BACKGROUND AND AIM: Upper gastrointestinal endoscopy is generally accepted as the gold standard for the clinical evaluation of gastric cancer (GC). However, the efficacy of endoscopic screening for asymptomatic GC remains controversial. The present study is designed to clarify the efficacy of endoscopic screening for the detection of early GC by investigating the clinicopathological features. METHODS: A total of 17 522 patients who had underwent endoscopic screening as a part of their annual health checkup at the Seirei Center for Health Promotion and Preventive Medicine between April 2002 and March 2006 were enrolled in this study. We investigated the clinicopathological findings of GC detected by endoscopy. Furthermore, in accordance with the screening interval at our center, patients with GC were categorized into two groups: group A, patients with repeated endoscopic screening within the last 2 years, and group B, patients without endoscopic screening within the last 2 years. RESULTS: Thirty-nine GC (mean age of patients: 62.2 +/- 8.0 years, 36 males and three females) were detected in total (0.22%). The proportion of early GC was 87.2%. Notable differences between groups A and B were not found in the rate of early GC (P = 0.6342). However, eight of 27 cases (29.6%) in group A were treated by endoscopic resection, but none in group B (P = 0.0344). In six of 26 cases (23.1%) in group A, the recorded images from the previous endoscopic examination indicated some macroscopic abnormalities at the same location, suggesting GC or premalignant lesions. CONCLUSION: Endoscopic screening is useful for detecting GC at the early stages, and repeated examinations at short-time intervals contribute to the detection of resectable lesions by endoscopy. Further studies are needed to decrease the false negative rate of endoscopic screening. PMID- 18397490 TI - Education and imaging. Gastrointestinal: Gastrointestinal histoplasmosis. PMID- 18397491 TI - Education and imaging. Hepatobiliary and pancreatic: Pancreatic ascites. PMID- 18397492 TI - Education and imaging. Gastrointestinal: Phlebosclerotic colitis. PMID- 18397493 TI - Education and imaging. Hepatobiliary and pancreatic: Systemic mastocytosis with ascites. PMID- 18397494 TI - Education and imaging. Gastrointestinal: The sign of Leser-Trelat. PMID- 18397495 TI - Primary jejunal adenocarcinoma as part of multiple primary cancers of the digestive tract. AB - Multiple primary cancers including small intestinal tumors are rare. We describe the first curative resection case of metachronous triple early cancers involving the jejunum as well as the stomach and esophagus. The patient had undergone total gastrectomy for a gastric adenocarcinoma and subsequent esophagectomy for an esophageal squamous cell carcinoma. A jejunal adenocarcinoma, the third primary, occurred at the blind stump of the jejunal limb of a Roux-en-Y esophagojejunostomy reconstructed previously. This tumor was removed by partial resection of the limb with the preservation of the esophagojejunostomy. The patient died from complications of acute pancreatitis 3 years after the last operation, without overt clinical signs of cancer recurrence. Immunohistochemistry of the specimen showed the increased expression of p53 and cyclin D1 proteins in all three cancers, suggesting their involvement in metachronous carcinogenesis in this case. Early diagnosis of each cancer was made possible by regular endoscopic follow-up and favorable anatomical location of each tumor, which were considered to allow less invasive surgery as well as to contribute to the favorable outcome. This case suggests the importance of regular surveillance for metachronous carcinogenesis, especially when the preceding cancers carry genetic abnormalities that may potentially increase the risk for subsequent carcinogenesis. PMID- 18397496 TI - Foot-and-mouth disease virus concentrations in products of animal origin. AB - Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals which can have devastating economic consequences. Maintaining an FMD-free status is a priority for non-endemic countries, which restrict importation of animals and animal products from countries in which the disease is present or sporadic, thus presenting a considerable barrier to international trade. This review examines the concentration of FMD virus in animal tissues during the viraemic stage of disease and in animal products derived from infected animals. PMID- 18397497 TI - Detection of endoparasites with zoonotic potential in dogs with gastrointestinal disease in the UK. AB - We report a substantial prevalence study in symptomatic pet dogs of important zoonotic parasitic enteric infections. A total of 4526 dogs which had a faecal sample submitted to a diagnostic laboratory in the UK between 2003 and 2005 were included in the study. The most common parasite was Giardia spp., which was found in 380/4526 dogs (8.4%, 95% CI 7.6-9.2%). Surprisingly, Cryptosporidium spp. infection was detected in only 29/4526 (0.6%, 95% CI 0.4-0.9%). Toxocara canis was found in 63/4526 dogs (1.4%; 95% CI 1.1-1.8%). Prevalence of Giardia (P < 0.001) was significantly higher in dogs <12 months of age, with nearly one-fifth of all symptomatic dogs under 6 months being infected with Giardia. Some seasonality was detected with a higher prevalence of Cryptosporidium oocyst shedding found from October to December. These data are of importance for veterinarians in judging the likelihood of enteric parasitic infection in an individual with clinical signs. Moreover, they provide information to direct future work in determining the risk to the human population from parasitic zoonoses of dogs. PMID- 18397498 TI - In silico comparative analysis of DNA and amino acid sequences for prion protein gene. AB - Genetic variability might contribute to species specificity of prion diseases in various organisms. In this study, structures of the prion protein gene (PRNP) and its amino acids were compared among species of which sequence data were available. Comparisons of PRNP DNA sequences among 12 species including human, chimpanzee, monkey, bovine, ovine, dog, mouse, rat, wallaby, opossum, chicken and zebrafish allowed us to identify candidate regulatory regions in intron 1 and 3' untranslated region (UTR) in addition to the coding region. Highly conserved putative binding sites for transcription factors, such as heat shock factor 2 (HSF2) and myocite enhancer factor 2 (MEF2), were discovered in the intron 1. In 3'-UTR, the functional sequence (ATTAAA) for nucleus-specific polyadenylation was found in all the analysed species. The functional sequence (TTTTTAT) for maturation-specific polyadenylation was identically observed only in ovine, and one or two nucleotide mismatches in the other species. A comparison of the amino acid sequences in 53 species revealed a large sequence identity. Especially the octapeptide repeat region was observed in all the species but frog and zebrafish. Functional changes and susceptibility to prion diseases with various isoforms of prion protein could be caused by numeric variability and conformational changes discovered in the repeat sequences. PMID- 18397499 TI - Evaluation of the pathogenicity and transmissibility of a chilean isolate of porcine reproductive and respiratory syndrome virus. AB - The objective of this study was to determine clinical features, shedding and transmission of a Chilean Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) strain upon experimental inoculation of 4-week-old pigs. Six groups of five animals each were used. The G1 (donor) group was inoculated with PRRSV, maintained in an isolation unit for 35 days, and sampled daily to determine shedding in mucosal secretions and faeces, viraemia and seroconversion. An uninfected control group (G6) was equally maintained and sampled under strict isolation. Four other groups (G2 to G5) were exposed to PRRSV via direct contact with G1 for 5-day periods in a staggered manner, throughout the 35-day period, and were later placed in an independent isolation unit to monitor infection status for 7 days. All the animals in G1 and G6 were killed at 35 days post inoculation (dpi) and the contact groups at 12 days post-contact (dpc). Samples were obtained from diverse organs for histopathological, immunohistochemical (IHC) and virological analysis. No clinical symptoms were evident in any group, except for a transient fever observed in G1. Histopathologically, all the animals of G1 had interstitial pneumonia, although scarce PRRSV-positive cells were detected in the lung using IHC. PRRSV-positive cells (IHC) were detected in the lymphoid tissue of all animals in infected groups, but especially in G3 and G4. Viraemia was detected in G1 (3-35 dpi) and in the all contact groups (5-12 dpc). Likewise, ranging from 3 to 19 dpi, PRRSV was detected in at least one animal from the tonsils and lungs in all infected groups, in nasal and ocular secretions, saliva or faeces. These results indicate that the donor group excreted infectious PRRSV and was able to transmit the infection to susceptible pigs. The critical shedding period was 7-19 dpi, during which, most likely, transmission took place. PMID- 18397500 TI - Association among results of serum ELISA, faecal culture and nested PCR on milk, blood and faeces for the detection of paratuberculosis in dairy cows. AB - Paratuberculosis is a chronic, infectious disease of ruminants that entails a serious concern for the cattle industry. One of the main issues relates to the efficiency of diagnosis of subclinically infected animals. The objective of this field study was to analyse the association among results of a serum enzyme-linked immunosorbent assays (ELISA), faecal culture and nested PCR tests on milk, blood and faeces for Mycobacterium avium subsp. paratuberculosis detection in dairy cows. Faeces, blood and milk samples were collected from 328 lactating dairy cows in four known infected herds. Results were analysed to determine associations and levels of agreement between pairs of tests. A total of 61 animals (18.6%) tested positive when all the tests were interpreted in parallel. The agreement between results in different pairs of tests was poor, slight and fair in two, five and three of the 10 possible combinations respectively. Faecal culture and faecal polymerase chain reaction (PCR) resulted in the highest kappa coefficient (0.39; fair agreement), with the lowest agreement being for ELISA and blood PCR (-0.036; poor agreement). Fisher's exact test resulted in statistically significant associations (P < or = 0.05) between the following test pairs: ELISA : faecal culture; ELISA : faecal PCR; milk PCR : faecal PCR, blood PCR : faecal PCR and faecal culture : faecal PCR. Enzyme-linked immunosorbent assays showed the highest complementary sensitivity values for all the possible two-test combinations, followed by faecal PCR. The combined use of ELISA and faecal PCR has the potential to increase the overall sensitivity for the diagnosis of paratuberculosis infection. PMID- 18397501 TI - Pathological, entomological, avian and meteorological investigation of a West Nile virus epidemic in a horse farm. AB - Pathological, entomological and avian investigations were conducted during the summer of 2002, in a horse farm that had four cases of West Nile virus (WNV) infection in horses. All the four horses had encephalitis and WNV infection was confirmed by RT-PCR and in situ hybridization procedure. Forty-seven per cent of house sparrows that resided on the farm were tested positive for WNV infection. Mosquitoes (98%Culex pipiens) collected by trapping at the farm, during this period were positive for WNV. The meteorological data for year 2002 were compared to previous 16 years. The precipitation and atmospheric temperature were found to be reduced and higher respectively, indicating a drier summer than the prior 16 years, which may have been a contributing factor for the outbreak. None of the horses on these premises had been vaccinated for WNV disease. PMID- 18397502 TI - Effect of natural exposure to vaccine-derived North American genotype PRRS virus on the serological response in naive pigs. AB - Diagnosis of porcine reproductive and respiratory syndrome virus (PRRSV) is often performed by serological testing, but ELISA does not differentiate between infections with wild-type or vaccine virus. Two attenuated live vaccines [European (EU) or North American (NA) genotype] are used. In addition to wild type isolates, vaccine or vaccine-derived viruses occur frequently. This is often not considered when the ELISA results are used to differentiate between epizootic and enzootic infections. In this study, an infection with the NA genotype vaccine derived virus was detected in two herds previously PRRSV negative and ELISA results [sample to positive (s/p) ratios] were analysed. The virus was identified by RT-PCR and nucleotide sequences of ORF5 had 97% (herd A) and 99% (herd B) identity with the genome of a ML PRRSV vaccine belonging to the NA genotype. Pigs of different age became positive with an average s/p ratio of 2.24 (A) and 1.18 (B). The data clearly demonstrate that spontaneous infection with a vaccine derived virus of the NA genotype induces ELISA s/p ratios similar to those induced by vaccination or by infection with wild-type virus. We conclude that for a correct interpretation of serological results the circulation of vaccine or vaccine-derived virus isolates has to be excluded by RT-PCR, even if vaccination is not ongoing. PMID- 18397503 TI - Transboundary disease management: the theory and the practice--the science and the politics. PMID- 18397504 TI - EFSA assessment of the risk of introducing foot and mouth disease into the EU and the reduction of this risk through interventions in infected countries: a review and follow-up. PMID- 18397505 TI - Incidence and distribution of foot-and-mouth disease in Asia, Africa and South America; combining expert opinion, official disease information and livestock populations to assist risk assessment. AB - Risk assessment procedures frequently require quantitative data on the prevalence of the disease in question. Although most countries are members of the World Organization for Animal Health (OIE), the importance attached to foot-and-mouth disease (FMD) reporting or surveillance for infection varies enormously between infected countries. There is a general consensus that FMD outbreaks in endemic countries are greatly under-reported, to a degree related either to the economic or the political development level of the country. This exploratory study was first undertaken by FAO, but thereafter extended and reviewed by the working group on FMD risk co-ordinated by the European Food Safety Authority (EFSA). The paper attempts to overcome the lack of reporting through using expert opinion to extrapolate incidence indices from countries considered to have 'representative' levels of FMD. These were combined with livestock density distributions to provide maps of prevalence indices, which were found to be highest in China (pigs), India (cattle), the Near East (small ruminants) and the Sahel (small ruminants and cattle). Similar patterns were found when weighted expert rankings of a range of additional ranked disease parameters were also produced, and then combined with susceptible animal densities to produce a weighted multi-species density. Results suggest that the methods can provide useful information at both national and sub-national resolution, even for countries for which quantitative FMD data is currently unavailable: two of the regions identified provide little or no data on a regular basis to the OIE and therefore may be overlooked if the level of officially reported FMD is only used. As the estimated prevalences are based on recent disease history and expert opinion, they are most likely to be inaccurate where FMD incursions are infrequent as a result of the preventive measures and geographical and trade isolation. This study, therefore, highlights the need for specific detailed country risk assessments where livestock trade is under consideration. Validating the approach including ground truthing, will require collaboration between a number of agencies and institutions, in critical countries, particularly those with high disease burdens that share borders or trade livestock with currently FMD-free nations. PMID- 18397506 TI - Incursions of foot-and-mouth disease virus into Europe between 1985 and 2006. AB - Foot-and-mouth disease (FMD) is one of the biggest threats to animal health in European countries. In the last 22 years (1985-2006), FMD has occurred 37 times in 14 European countries. Serotype O was most frequently involved in these outbreaks followed by A, C and Asia 1. Sometimes, epidemics were very limited and at other times, they were the cause of devastating economic losses. In most cases (22/37), the origin of the outbreaks could not be determined. For some of these outbreaks, however, routes of introduction and spread were identified through epidemiological inquiries. Moreover, in some cases, the origin of the virus was also traced by phylogenetic analysis of the partial or complete sequences of VP1 genes. Lessons learned from the outbreaks are still useful as most of the same risk factors persist. However, efforts made by FMD-free countries to help those where the disease is endemic are a valuable strategy for the reduction of the global risk. The present and the future potential sources of FMD infection need to be identified to best focus European efforts. PMID- 18397507 TI - The importance of quality assurance/quality control of diagnostics to increase the confidence in global foot-and-mouth disease control. AB - The last decade international trade in animals and animal products was liberated and confidence in this global trade can increase only if appropriate control measures are applied. As foot-and-mouth disease (FMD) diagnostics will play an essential role in this respect, the Food and Agriculture Organization European Commission for the Control of Foot-and-Mouth Disease (EUFMD) co-ordinates, in collaboration with the European Commission, several programmes to increase the quality of FMD diagnostics. A quality assurance (QA) system is deemed essential for laboratories involved in certifying absence of FMDV or antibodies against the virus. Therefore, laboratories are encouraged to validate their diagnostic tests fully and to install a continuous quality control (QC) monitoring system. Knowledge of performance characteristics of diagnostics is essential to interpret results correctly and to calculate sample rates in regional surveillance campaigns. Different aspects of QA/QC of classical and new FMD virological and serological diagnostics are discussed in respect to the EU FMD directive (2003/85/EC). We recommended accepting trade certificates only from laboratories participating in international proficiency testing on a regular basis. PMID- 18397508 TI - FMD vaccines: reflections on quality aspects for applicability in European disease control policy. AB - Most foot-and-mouth disease (FMD) vaccines used around the world are inactivated vaccines for prophylactic or emergency use, generally manufactured by the same basic methodology outlined in the OIE Manual and, for Europe, in the European Pharmacopoeia, and for the EU Member States in compliance with Directive 2001/82/EC of the European Parliament and of the Council of 6 November 2001 on the Community code relating to veterinary medicinal products as amended by Directive 2004/28/EC. Most of the requirements that apply to all immunological veterinary medicinal products apply equally to FMD vaccines. There are, however, some unique features of the disease and vaccines used against it that require a different approach to fulfil the requirements of the relevant legislation, if a vaccinate-to-live policy will be applied with 'authorized' vaccines. Several aspects of vaccine efficacy and safety are elaborated with emphasis on quality assurance/quality control (QA/QC). The purity of the vaccine in respect of the presence of non-structural protein antibodies could be checked indirectly by serology after vaccination. The viability of a vaccine bank approach was greatly aided by the principle of storing inactivated concentrated FMD viral antigen (Ag) over liquid nitrogen for subsequent formulation into vaccine. A worldwide Ag bank network might be an option for the far future and a solution to the problem of covering many different FMDV serotypes and strains. The producers should respect the strict FMD biosecurity rules worked out by the FAO EUFMD and described in Council Directive 2003/85/EC. Making the experience related to vaccine QA/QC available to all countries will reduce the risk of an FMD outbreak within these countries and consequently will reduce the FMD risk around the world. PMID- 18397509 TI - Epidemiological patterns of foot-and-mouth disease worldwide. AB - Foot-and-Mouth Disease (FMD) is a clinical syndrome in animals due to FMD virus that exists in seven serotypes, whereby recovery from one sero-type does not confer immunity against the other six. So when considering intervention strategies in endemic settings, it is important to take account of the characteristics of the different serotypes in different ecological systems. FMD serotypes are not uniformly distributed in the regions of the world where the disease still occurs. For example, the cumulative incidence of FMD serotypes show that six of the seven serotypes of FMD (O, A, C, SAT-1, SAT-2, SAT-3) have occurred in Africa, while Asia contends with four sero-types (O, A, C, Asia-1), and South America with only three (O, A, C). Periodically there have been incursions of Types SAT-1 and SAT-2 from Africa into the Middle East. This paper describes the global dynamics for the seven sero-types and attempts to define FMD epidemiological clusters in the different regions of the world. These have been described on a continent by continent basis. The review has reaffirmed that the movement of infected animals is the most important factor in the spread of FMD within the endemically infected regions. It also shows that the eco-system based approach for defining the epidemiological patterns of FMD in endemic, which was originally described in South America, can apply readily to other parts of the world. It is proposed that any coordinated regional or global strategy for FMD control should be based on a sound epidemiological assessment of the incidence and distribution of FMD, identifying risk sources as either primary or secondary endemic eco-systems. PMID- 18397510 TI - Planning for the progressive control of foot-and-mouth disease worldwide. AB - In the wake of on-going successful programmes for global eradication of rinderpest and the current effort to contain the spread of avian influenza, the progressive world-wide control of FMD must be regarded as a major contribution to the international public good. FMD is the single most animal disease constraint to international trade in animal products. Its control is relevant, on the one hand, to protecting the livestock industries of industrialised countries and, on the other, to the livelihoods and income generation of developing countries, where, as a general rule, FMD continues to be endemic. The strategy that is advocated in this paper is one that is based on progressive risk reduction of FMD in the context of progressive market access of livestock commodities from developing countries. It is suggested that FMD control should be linked to improvement in livelihoods of livestock dependent communities in the FMD endemic settings. It is expected that this in turn will lead to increasing demand for effective national veterinary services and disease surveillance. This strategy has also taken lessons from the global rinderpest eradication programme and regional FMD control programmes in Europe and South America. The strategy that is advocated for the progressive control of FMD in the endemic settings is based on a seven stage process within a horizon of about 30 years, namely: (1) Assessing and defining national FMD status; (2) instituting vaccination and movement control; (3) suppressing virus transmission to achieve absence of clinical disease; (4) achieving freedom from FMD with vaccination in accordance with the OIE standards; (5) achieving freedom from FMD without vaccination in accordance with the OIE standards; (6) extending FMD free zones; and (7) maintaining FMD Freedom. Concomitant with progressive FMD control, there needs be the encouragement of such risk reduction measures as in-country commodity processing in order to encourage regulated trade in livestock commodities without unduly increasing the risk of disease spread. Finally, the progressive control of FMD should also be seen as part of reducing the overall, world-wide threat of infectious diseases to human health and economic development. PMID- 18397511 TI - Non-systemic metamorphosis in male millipede appendages: long delayed, reversible effect of an early localized positional marker? AB - BACKGROUND: The development of specialized appendages involved in sperm transfer in the males of julid millipedes is an extreme case of specialized, complex structures differentiating in a very advanced phase of post-embryonic development. Here, a non-systemic metamorphosis affects the external morphology and the internal anatomy of a trunk double segment only. PRESENTATION OF THE HYPOTHESIS: We hypothesize that during early (possibly embryonic) development a segmental marker is produced that remains unexploited throughout late embryonic and early post-embryonic development, until, activated by a systemic signal, it finally determines the release of a segmentally localized but anatomically major change. TESTING THE HYPOTHESIS: Key to testing the hypothesis are (1) the identification of both the putative segmental marker involved in the localization of the legs to be eventually metamorphosed into gonopods and the systemic signal activating it, (2) the identification of the cell population from which the gonopods are built, and (3) a longitudinal study of the marker's expression throughout late embryonic and, possibly, post-embryonic development. IMPLICATIONS OF THE HYPOTHESIS: Proving the validity of this hypothesis would demonstrate the existence of a cryptic developmental module that will be activated only months, or years, after it has been first laid down during early development. This study also opens a window onto the very poorly explored domain of late expression of developmental genes and molecular control of late developmental events. PMID- 18397512 TI - Hospitalizations associated with rotavirus gastroenteritis in Spain, 2001-2005. AB - BACKGROUND: This study aims to describe and analyze hospital admissions in Spain due to rotavirus infections among children aged 5 years or under during the period 2001-2005, along with the associated health cost. METHODS: To update estimates of rotavirus hospitalizations rates in Spain, we conducted a retrospective study of 5 years of national hospitalization data associated with acute gastroenteritis using the Minimum Basic Data Set. RESULTS: During the study period, a total of 17.1% of all admissions due to acute gastroenteritis of any etiology in children aged < or = 5 years were attributable to rotavirus infection as determined by the rotavirus-specific International Classification of Diseases, ninth revision, Clinical Modification code. A mean incidence of 135 hospital admissions attributable to rotavirus per 100,000 children aged < or = 5 years was found. Hospitalizations associated with rotavirus had a marked winter-time seasonality. The estimated cost of hospital admission attributable to rotavirus has risen from 3 million euros estimated for 2001 to almost 7 million euros estimated in 2005. CONCLUSION: Rotavirus gastroenteritis remains an important cause of hospitalizations in Spanish children, mostly during the winter season. PMID- 18397513 TI - Evidence of scrapie transmission via milk. AB - BACKGROUND: The risk of scrapie infection increases with increased duration and proximity of contact between sheep at lambing. Scrapie infectivity has not been detected in milk but cellular prion protein, the precursor of disease-associated prion protein PrPd, has been found in milk from ruminants. To determine whether milk is able to transmit scrapie, 18 lambs with a prion protein genotype associated with high susceptibility to scrapie (VRQ/VRQ) were fed milk from twelve scrapie-affected ewes of the same genotype, and 15 VRQ/VRQ sheep reared on scrapie-free dams served as controls. RESULTS: Three lambs fed milk from scrapie affected ewes were culled due to intercurrent diseases at 43, 44 and 105 days of age respectively, and PrPd was detected in the distal ileum of the first two lambs, whilst PrPd was not found in lymphoreticular tissues in the third lamb. A control lamb, housed in a separate pen and culled at 38 days of age, was also negative for PrPd in a range of tissues. Samples of recto-anal mucosa associated lymphoid tissue collected from the remaining 15 live lambs at seven months of age (between five to seven months after mixing) were positive for PrPd in the scrapie milk recipients, whereas PrPd was not detected in the remaining 14 controls at that time. A subsequent sample collected from control lambs revealed PrPd accumulation in two of five lambs eight months after mixing with scrapie milk recipients suggestive of an early stage of infection via lateral transmission. By contrast, the control sheep housed in the same building but not mixed with the scrapie milk recipients were still negative for PrPd. CONCLUSION: The presence of PrPd in distal ileum and rectal mucosa indicates transmission of scrapie from ewe to lamb via milk (or colostrum) although it is not yet clear if such cases would go on to develop clinical disease. The high level of infection in scrapie-milk recipients revealed by rectal mucosal testing at approximately seven months of age may be enhanced or supplemented by intra-recipient infection as these lambs were mixed together after feeding with milk from scrapie-affected ewes and we also observed lateral transmission from these animals to lambs weaned from scrapie-free ewes. PMID- 18397514 TI - Diversity and seasonal dynamics of bacterial community in indoor environment. AB - BACKGROUND: We spend most of our lives in indoor environments and are exposed to microbes present in these environments. Hence, knowledge about this exposure is important for understanding how it impacts on human health. However, the bacterial flora in indoor environments has been only fragmentarily explored and mostly using culture methods. The application of molecular methods previously utilised in other environments has resulted in a substantial increase in our awareness of microbial diversity. RESULTS: The composition and dynamics of indoor dust bacterial flora were investigated in two buildings over a period of one year. Four samples were taken in each building, corresponding to the four seasons, and 16S rDNA libraries were constructed. A total of 893 clones were analysed and 283 distinct operational taxonomic units (OTUs) detected among them using 97% sequence similarity as the criterion. All libraries were dominated by Gram-positive sequences, with the most abundant phylum being Firmicutes. Four OTUs having high similarity to Corynebacterium-, Propionibacterium-, Streptococcus- and Staphylococcus- sequences were present in all samples. The most abundant of the Gram-negative OTUs were members of the family Sphingomonadaceae, followed by Oxalobacteraceae, Comamonadaceae, Neisseriaceae and Rhizobiaceae. The relative abundance of alpha- and betaproteobacteria increased slightly towards summer at the expense of firmicutes. The proportion of firmicutes and gammaproteobacteria of the total diversity was highest in winter and that of actinobacteria, alpha- and betaproteobacteria in spring or summer, whereas the diversity of bacteroidetes peaked in fall. A statistical comparison of the libraries revealed that the bacterial flora of the two buildings differed during all seasons except spring, but differences between seasons within one building were not that clear, indicating that differences between the buildings were greater than the differences between seasons. CONCLUSION: This work demonstrated that the bacterial flora of indoor dust is complex and dominated by Gram-positive species. The dominant phylotypes most probably originated from users of the building. Seasonal variation was observed as proportional changes of the phyla and at the species level. The microflora of the two buildings investigated differed statistically and differences between the buildings were more pronounced than differences between seasons. PMID- 18397515 TI - Costs and benefits of multiple resistance to insecticides for Culex quinquefasciatus mosquitoes. AB - BACKGROUND: The evolutionary dynamics of xenobiotic resistance depends on how resistance mutations influence the fitness of their bearers, both in the presence and absence of xenobiotic selection pressure. In cases of multiple resistance, these dynamics will also depend on how individual resistance mutations interact with one another, and on the xenobiotics applied against them. We compared Culex quinquefasciatus mosquitoes harbouring two resistance alleles ace-1R and KdrR (conferring resistance to carbamate and pyrethroid insecticides, respectively) to mosquitoes bearing only one of the alleles, or neither allele. Comparisons were made in environments where both, only one, or neither type of insecticide was present. RESULTS: Each resistance allele was associated with fitness costs (survival to adulthood) in an insecticide-free environment, with the costs of ace 1R being greater than for KdrR. However, there was a notable interaction in that the costs of harbouring both alleles were significantly less than for harbouring ace-1R alone. The two insecticides combined in an additive, synergistic and antagonistic manner depending on a mosquito's resistance status, but were not predictable based on the presence/absence of either, or both mutations. CONCLUSION: Insecticide resistance mutations interacted to positively or negatively influence a mosquito's fitness, both in the presence or absence of insecticides. In particular, the presence of the KdrR mutation compensated for the costs of the ace-1R mutation in an insecticide-free environment, suggesting the strength of selection in untreated areas would be less against mosquitoes resistant to both insecticides than for those resistant to carbamates alone. Additional interactions suggest the dynamics of resistance will be difficult to predict in populations where multiple resistance mutations are present or that are subject to treatment by different xenobiotics. PMID- 18397516 TI - Prevention of infectious tick-borne diseases in humans: Comparative studies of the repellency of different dodecanoic acid-formulations against Ixodes ricinus ticks (Acari: Ixodidae). AB - BACKGROUND: Ticks of the species Ixodes ricinus are the main vectors of Lyme Borreliosis and Tick-borne Encephalitis - two rapidly emerging diseases in Europe. Repellents provide a practical means of protection against tick bites and can therefore minimize the transmission of tick-borne diseases. We developed and tested seven different dodecanoic acid (DDA)-formulations for their efficacy in repelling host-seeking nymphs of I. ricinus by laboratory screening. The ultimately selected formulation was then used for comparative investigations of commercially available tick repellents in humans. METHODS: Laboratory screening tests were performed using the Moving-object (MO) bioassay. All test formulations contained 10% of the naturally occurring active substance DDA and differed only in terms of the quantitative and qualitative composition of inactive ingredients and fragrances. The test procedure used in the human bioassays is a modification of an assay described by the U.S. Environmental Protection Agency and recommended for regulatory affairs. Repellency was computed using the equation: R = 100 - NR/N x 100, where NR is the number of non-repelled ticks, and N is the respective number of control ticks. All investigations were conducted in a controlled laboratory environment offering standardized test conditions. RESULTS: All test formulations strongly repelled nymphs of I. ricinus (100-81% protection) as shown by the MO-bioassay. The majority of ticks dropped off the treated surface of the heated rotating drum that served as the attractant (1 mg/cm2 repellent applied). The 10% DDA-based formulation, that produced the best results in laboratory screening, was as effective as the coconut oil-based reference product. The mean protection time of both preparations was generally similar and averaged 8 hours.Repellency investigations in humans showed that the most effective 10% DDA based formulation (~1.67 mg/cm2 applied) strongly avoided the attachment of I. ricinus nymphs and adults for at least 6 hours. The test repellent always provided protection (83-63%) against I. ricinus nymphs equivalent to the natural coconut oil based reference product and a better protection (88-75%) against adult ticks than the synthetic Icaridin-containing reference repellent. CONCLUSION: We found that the 10% DDA-based formulation (ContraZeck(R)) is an easily applied and very effective natural repellent against I. ricinus ticks. By reducing the human-vector contact the product minimises the risk of transmission of tick-borne diseases in humans. PMID- 18397517 TI - Evolutionary potentials: structure specific knowledge-based potentials exploiting the evolutionary record of sequence homologs. AB - We introduce a new type of knowledge-based potentials for protein structure prediction, called 'evolutionary potentials', which are derived using a single experimental protein structure and all three-dimensional models of its homologous sequences. The new potentials have been benchmarked against other knowledge-based potentials, resulting in a significant increase in accuracy for model assessment. In contrast to standard knowledge-based potentials, we propose that evolutionary potentials capture key determinants of thermodynamic stability and specific sequence constraints required for fast folding. PMID- 18397518 TI - Myocardial ischemia in the absence of epicardial coronary artery disease in Friedreich's ataxia. AB - We present the first in vivo detection of microvascular abnormality in a patient with Friedreich's ataxia (FA) without epicardial coronary artery disease using cardiac magnetic resonance (CMR). The patient had exertional chest pain and dyspnea prompting referral for cardiac evaluation. These symptoms were reproduced during intravenous adenosine infusion, and simultaneous first-pass perfusion imaging showed a significant subendocardial defect; both symptoms and perfusion deficit were absent at rest. Epicardial coronaries were free of disease by invasive angiography; together, these findings support the notion of impaired myocardial perfusion reserve in FA. PMID- 18397520 TI - The impact of the treaty basis on health policy legislation in the European Union: a case study on the tobacco advertising directive. AB - BACKGROUND: The Europe Against Cancer programme was initiated in the late 1980s, recognising, among other risk factors, the problematic relationship between tobacco use and cancer. In an attempt to reduce the number of smokers in the European Community, the European Commission proposed a ban on tobacco advertising. The question of why it took over ten years of negotiating before the EU adopted a policy measure that could in fact improve the health situation in the Community, can only be answered by focusing on politics. METHODS: We used an actor-centred institutionalist approach, focusing on the strategic behaviour of the major actors involved. We concentrated our analysis on the legal basis as an important institution and evaluated how the absence of a proper legal basis for public health measures in the Treaties influenced policy-making, framing the discussion in market-making versus market-correcting policy interventions. For our analysis, we used primary and secondary sources, including policy documents, communications and press releases. We also conducted 9 semi-structured interviews. RESULTS: The ban on tobacco advertising was, in essence, a public health measure. The Commission used its agenda-setting power and framed the market-correcting proposal in market-making terms. The European Parliament and the Council of Ministers then used the discussion on the legal basis as a vehicle for real political controversies. After adoption of the ban on tobacco advertising, Germany appealed to the European Court of Justice, which annulled the ban but also offered suggestions for a possible solution with article 100a as the legal basis. CONCLUSION: The whole market-making versus market-correcting discussion is related to a broader question, namely how far European health regulation can go in respect to the member states. In fact, the policy-making process of a tobacco advertising ban, as described in this paper, is related to the 'constitutional' foundation of health policy legislation in the Community. The absence of a clear-cut legal basis for health policies does not imply that the EU's impact on health is negligible. In the case of tobacco-control measures, the creative use of other Treaty bases has resulted in significant European action in the field of public health. PMID- 18397519 TI - Prioritisation of patients on waiting lists for hip and knee arthroplasties and cataract surgery: Instruments validation. AB - BACKGROUND: Prioritisation instruments were developed for patients on waiting list for hip and knee arthroplasties (AI) and cataract surgery (CI). The aim of the study was to assess their convergent and discriminant validity and inter observer reliability. METHODS: Multicentre validation study which included orthopaedic surgeons and ophthalmologists from 10 hospitals. Participating doctors were asked to include all eligible patients placed in the waiting list for the procedures under study during the medical visit. Doctors assessed patients' priority through a visual analogue scale (VAS) and administered the prioritisation instrument. Information on socio-demographic data and health related quality of life (HRQOL) (HUI3, EQ-5D, WOMAC and VF-14) was obtained through a telephone interview with patients. The correlation coefficients between the prioritisation instrument score and VAS and HRQOL were calculated. For the reliability study a self-administered questionnaire, which included hypothetic patients' scenarios, was sent via postal mail to the doctors. The priority of these scenarios was assessed through the prioritisation instrument. The intraclass correlation coefficient (ICC) between doctors was calculated. RESULTS: Correlations with VAS were strong for the AI (0.64, CI95%: 0.59-0.68) and for the CI (0.65, CI95%: 0.62-0.69), and moderate between the WOMAC and the AI (0.39, CI95%: 0.33-0.45) and the VF-14 and the CI (0.38, IC95%: 0.33-0.43). The results of the discriminant analysis were in general as expected. Inter-observer reliability was 0.79 (CI95%: 0.64-0.94) for the AI, and 0.79 (CI95%: 0.63-0.95) for the CI. CONCLUSION: The results show acceptable validity and reliability of the prioritisation instruments in establishing priority for surgery. PMID- 18397521 TI - Ovarian cancer risk in Polish BRCA1 mutation carriers is not associated with the prohibitin 3' untranslated region polymorphism. AB - BACKGROUND: The variable penetrance of ovarian cancer in BRCA1 mutation carriers suggests that other genetic or environmental factors modify disease risk. The C to T transition in the 3' untranslated region of the prohibitin (PHB) gene alters mRNA function and has recently been shown to be associated with hereditary breast cancer risk in Polish women harbouring BRCA1 mutations. METHODS: To investigate whether the PHB 3'UTR polymorphism also modifies hereditary ovarian cancer risk, we performed a case-control study among Polish women carrying one of the three common founder mutations (5382insC, 300 T > G, 4154delA) including 127 ovarian cases and 127 unaffected controls who had both breasts and ovaries intact. Controls were matched to cases by year of birth and BRCA1 mutation. Genotyping analysis was performed using PCR-based restriction fragment length polymorphism analysis. Odds ratios (OR) were calculated using conditional and penalized univariable and multivariable logistic regression. RESULTS: A comparison of the genotype frequencies between cases and controls revealed no association of the PHB 3'UTR _CT+TT genotypes with ovarian cancer risk (ORadj 1.34; 95% CI, 0.59 3.11). CONCLUSION: Our data suggest that the PHB 3'UTR polymorphism does not modify ovarian cancer risk in women carrying one of the three Polish BRCA1 founder mutations. PMID- 18397522 TI - Dietary carbohydrate restriction in type 2 diabetes mellitus and metabolic syndrome: time for a critical appraisal. AB - Current nutritional approaches to metabolic syndrome and type 2 diabetes generally rely on reductions in dietary fat. The success of such approaches has been limited and therapy more generally relies on pharmacology. The argument is made that a re-evaluation of the role of carbohydrate restriction, the historical and intuitive approach to the problem, may provide an alternative and possibly superior dietary strategy. The rationale is that carbohydrate restriction improves glycemic control and reduces insulin fluctuations which are primary targets. Experiments are summarized showing that carbohydrate-restricted diets are at least as effective for weight loss as low-fat diets and that substitution of fat for carbohydrate is generally beneficial for risk of cardiovascular disease. These beneficial effects of carbohydrate restriction do not require weight loss. Finally, the point is reiterated that carbohydrate restriction improves all of the features of metabolic syndrome. PMID- 18397523 TI - DNA damage and cytotoxicity in type II lung epithelial (A549) cell cultures after exposure to diesel exhaust and urban street particles. AB - BACKGROUND: Exposure to air pollution particles has been acknowledged to be associated with excess generation of oxidative damage to DNA in experimental model systems and humans. The use of standard reference material (SRM), such as SRM1650 and SRM2975, is advantageous because experiments can be reproduced independently, but exposure to such samples may not mimic the effects observed after exposure to authentic air pollution particles. This study was designed to compare the DNA oxidizing effects of authentic street particles with SRM1650 and SRM2975. The authentic street particles were collected at a traffic intensive road in Copenhagen, Denmark. RESULTS: All of the particles generated strand breaks and oxidized purines in A549 lung epithelial cells in a dose-dependent manner and there were no overt differences in their potency. The exposures also yielded dose-dependent increase of cytotoxicity (as lactate dehydrogenase release) and reduced colony forming ability with slightly stronger cytotoxicity of SRM1650 than of the other particles. In contrast, only the authentic street particles were able to generate 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in calf thymus DNA, which might be due to the much higher level of transition metals. CONCLUSION: Authentic street particles and SRMs differ in their ability to oxidize DNA in a cell-free environment, whereas cell culture experiments indicate that the particle preparations elicit a similar alteration of the level of DNA damage and small differences in cytotoxicity. Although it cannot be ruled out that SRMs and authentic street particles might elicit different effects in animal experimental models, this study indicates that on the cellular level, SRM1650 and SRM2975 are suitable surrogate samples for the study of authentic street particles. PMID- 18397525 TI - The statistical analysis of a clinical trial when a protocol amendment changed the inclusion criteria. AB - BACKGROUND: Sometimes, protocol amendments that change the inclusion and exclusion criteria are required in clinical trials. Then, the patient populations before and after the amendment may differ. METHODS: We propose to perform separate statistical tests for the different phases, i.e. for the patients recruited before and after the amendment, and to combine the tests using Fisher's combination test. After a significant combination test a multiple testing procedure can be applied to identify the phase(s) to which a proof of efficacy refers. We assume that the amendment(s) are not based on any type of unblinded data. The proposed method is investigated within a simulation study. RESULTS: The proposed combination approach is superior to the 'naive' strategy to ignore the differences between the phases and pooling the data to perform just one statistical test. This superiority disappears when there are hardly any differences between the two phases. CONCLUSION: When one or more protocol amendments change the inclusion and exclusion criteria, one should realize that the populations may differ. In this case, separate tests for the different phases together with a combination test are a powerful method that can be applied in a variety of settings. The (first) amendment should specify the combination test to be applied in order to combine the different phases. PMID- 18397524 TI - Imported malaria in a cosmopolitan European city: a mirror image of the world epidemiological situation. AB - BACKGROUND: International travel and migration have been related with an increase of imported malaria cases. There has been considerable immigration to Barcelona from low-income countries (LIC) in recent years. The objective is to describe the epidemiology and to determine the trends of the disease in Barcelona. METHODS: Analysis of the cases notified among city residents between 1989 and 2005. Patients were classified as: tourists, voluntary workers, resident immigrants (visiting friends and relatives, VFR) and recently arrived immigrants. An analysis was conducted using the chi2 test and comparison of means. As a measure of association we calculated the Relative Risk (RR) and Odds Ratio (OR) with a Confidence Interval of 95% (CI) and carried out a trends analysis. RESULTS: Of the total of 1,579 imported cases notified, 997 (63.1%) lived in Barcelona city, and 55.1% were male. The mean age of patients was 32.7 years. The incidence increased from 2.4 cases/100,000 in 1989 to 3.5 cases/100,000 in 2005 (RR 1.46 CI:1.36-1.55). This increase was not statistically significant (trends analysis, p = 0.36). In terms of reason for travelling, 40.7% were VFR, 33.6% tourists, 12.1% voluntary workers and 13.6% were recently arrived immigrants. The most frequent species found was Plasmodium falciparum (71.3%), mainly in visitors to Africa (OR = 2.3, CI = 1.7-3.2). The vast majority (82.2%) had had some contact with Africa (35.9% with Equatorial Guinea, a Spanish ex-colony) and 96.6% had not completed chemoprophylaxis. Six deaths were observed, all tourists who had travelled to Africa and not taken chemoprophylaxis (3.9% fatality rate). CONCLUSION: Over the period studied there is an increase in malaria incidence, however the trend is not statistically significant. Lack of chemoprophylaxis compliance and the association between Africa and P. falciparum are very clear in the imported cases. Most of the patients with malaria did not take chemoprophylaxis. PMID- 18397526 TI - Natural selection of protein structural and functional properties: a single nucleotide polymorphism perspective. AB - BACKGROUND: The rates of molecular evolution for protein-coding genes depend on the stringency of functional or structural constraints. The Ka/Ks ratio has been commonly used as an indicator of selective constraints and is typically calculated from interspecies alignments. Recent accumulation of single nucleotide polymorphism (SNP) data has enabled the derivation of Ka/Ks ratios for polymorphism (SNP A/S ratios). RESULTS: Using data from the dbSNP database, we conducted the first large-scale survey of SNP A/S ratios for different structural and functional properties. We confirmed that the SNP A/S ratio is largely correlated with Ka/Ks for divergence. We observed stronger selective constraints for proteins that have high mRNA expression levels or broad expression patterns, have no paralogs, arose earlier in evolution, have natively disordered regions, are located in cytoplasm and nucleus, or are related to human diseases. On the residue level, we found higher degrees of variation for residues that are exposed to solvent, are in a loop conformation, natively disordered regions or low complexity regions, or are in the signal peptides of secreted proteins. Our analysis also revealed that histones and protein kinases are among the protein families that are under the strongest selective constraints, whereas olfactory and taste receptors are among the most variable groups. CONCLUSION: Our study suggests that the SNP A/S ratio is a robust measure for selective constraints. The correlations between SNP A/S ratios and other variables provide valuable insights into the natural selection of various structural or functional properties, particularly for human-specific genes and constraints within the human lineage. PMID- 18397527 TI - The development and deployment of Common Data Elements for tissue banks for translational research in cancer - an emerging standard based approach for the Mesothelioma Virtual Tissue Bank. AB - BACKGROUND: Recent advances in genomics, proteomics, and the increasing demands for biomarker validation studies have catalyzed changes in the landscape of cancer research, fueling the development of tissue banks for translational research. A result of this transformation is the need for sufficient quantities of clinically annotated and well-characterized biospecimens to support the growing needs of the cancer research community. Clinical annotation allows samples to be better matched to the research question at hand and ensures that experimental results are better understood and can be verified. To facilitate and standardize such annotation in bio-repositories, we have combined three accepted and complementary sets of data standards: the College of American Pathologists (CAP) Cancer Checklists, the protocols recommended by the Association of Directors of Anatomic and Surgical Pathology (ADASP) for pathology data, and the North American Association of Central Cancer Registry (NAACCR) elements for epidemiology, therapy and follow-up data. Combining these approaches creates a set of International Standards Organization (ISO) - compliant Common Data Elements (CDEs) for the mesothelioma tissue banking initiative supported by the National Institute for Occupational Safety and Health (NIOSH) of the Center for Disease Control and Prevention (CDC). METHODS: The purpose of the project is to develop a core set of data elements for annotating mesothelioma specimens, following standards established by the CAP checklist, ADASP cancer protocols, and the NAACCR elements. We have associated these elements with modeling architecture to enhance both syntactic and semantic interoperability. The system has a Java based multi-tiered architecture based on Unified Modeling Language (UML). RESULTS: Common Data Elements were developed using controlled vocabulary, ontology and semantic modeling methodology. The CDEs for each case are of different types: demographic, epidemiologic data, clinical history, pathology data including block level annotation, and follow-up data including treatment, recurrence and vital status. The end result of such an effort would eventually provide an increased sample set to the researchers, and makes the system interoperable between institutions. CONCLUSION: The CAP, ADASP and the NAACCR elements represent widely established data elements that are utilized in many cancer centers. Herein, we have shown these representations can be combined and formalized to create a core set of annotations for banked mesothelioma specimens. Because these data elements are collected as part of the normal workflow of a medical center, data sets developed on the basis of these elements can be easily implemented and maintained. PMID- 18397528 TI - The economic impact of ME/CFS: individual and societal costs. AB - BACKGROUND: ME/CFS is characterized by debilitating fatigue in addition to other physical and cognitive symptoms. It is estimated to affect over 800,000 adults in the U.S. ME/CFS often results in diminished functionality and increased economic impact. The economic impact of an illness is generally divided into two categories: direct and indirect costs. Despite high prevalence rates and the disabling nature of the illness, few studies have examined the costs of ME/CFS at the individual and societal level. In fact, of the four studies examining the economic impact of ME/ME/CFS only two used a U. S. sample. The current study used community and tertiary samples to examine the direct costs of ME/CFS. METHODS: Using archival data, Study 1 examined the direct cost of ME/CFS in a community based sample in Chicago. Study 2 estimated the direct cost of ME/CFS in a tertiary sample in Chicago. Both Study 1 and Study 2 assessed direct costs using office visit costs, medical test costs, and medication costs. RESULTS: For Study 1, the annual direct total cost per ME/CFS patient was estimated to be $2,342, with the total annual direct cost of ME/CFS to society being approximately $2 billion. In Study 2, the annual direct was estimated to be $8,675 per ME/CFS patient, with the total annual direct cost of ME/CFS to society being approximately $7 billion. CONCLUSION: Using ME/CFS prevalence data of 0.42 and indirect costs estimates from Reynolds et al. (2004), the direct and indirect cost of ME/CFS to society was estimated to be $18,677,912,000 for the community sample and $23,972,300,000 for the tertiary sample. These findings indicate that whether or not individuals are recruited from a community or tertiary sample, ME/CFS imposes substantial economic costs. PMID- 18397529 TI - Genetic and dietary salt contributors to insulin resistance in Dahl salt sensitive (S) rats. AB - Insulin resistance has been extensively investigated during the past decade because of its proposed role in initiating a cluster of cardiovascular risk factors including hypertension. Insulin resistance is an inherited genetic trait that precedes hypertension in Dahl salt-sensitive (S) rats, and is not present in Dahl salt-resistant (R) rats. Owing to the co-existence of insulin resistance and salt sensitivity of blood pressure in Dahl S, but not R rats, Dahl S rats are used to elucidate the role of dietary salt as a potential link in exacerbating both phenotypes (insulin resistance and salt sensitivity). In light of available data, examining the impact of dietary salt on insulin resistance in Dahl S rats in terms of salt concentration and duration of exposure helps answer the following question: What percentage of dietary salt and for what duration of exposure would we expect an enhanced insulin resistance in Dahl S rats? This commentary gathers all available research done on insulin resistance in Dahl S rats in an attempt to unravel dietary salt contribution to insulin resistance in Dahl S rats. PMID- 18397530 TI - Development of an instrument to assess the impact of an enhanced experiential model on pharmacy students' learning opportunities, skills and attitudes: a retrospective comparative-experimentalist study. AB - BACKGROUND: Pharmacy schools across North America have been charged to ensure their students are adequately skilled in the principles and practices of pharmaceutical care. Despite this mandate, a large percentage of students experience insufficient opportunities to practice the activities, tasks and processes essential to pharmaceutical care. The objective of this retrospective study of pharmacy students was to: (1) as "proof of concept", test the overall educational impact of an enhanced advanced pharmacy practice experiential (APPE) model on student competencies; (2) develop an instrument to measure students' and preceptors' experiences; and (3) assess the psychometric properties of the instrument. METHODS: A comparative-experimental design, using student and preceptor surveys, was used to evaluate the impact of the enhanced community based APPE over the traditional APPE model. The study was grounded in a 5-stage learning model: (1) an enhanced learning climate leads to (2) better utilization of learning opportunities, including (3) more frequent student/patient consultation, then to (4) improved skills acquisition, thence to (5) more favorable attitudes toward pharmaceutical care practice. The intervention included a one-day preceptor workshop, a comprehensive on-site student orientation and extending the experience from two four-week experiences in different pharmacies to one eight-week in one pharmacy. RESULTS: The 35 student and 38 preceptor survey results favored the enhanced model; with students conducting many more patient consultations and reporting greater skills improvement. In addition, the student self-assessment suggested changes in attitudes favoring pharmaceutical care principles. Psychometric testing showed the instrument to be sensitive, valid and reliable in ascertaining differences between the enhanced and traditional arms. CONCLUSION: The enhanced experiential model positively affects learning opportunities and competency acquisition, as measured by a new instrument showing sound psychometric properties. PMID- 18397531 TI - The L1-type cell adhesion molecule Neuroglian is necessary for maintenance of sensory axon advance in the Drosophila embryo. AB - BACKGROUND: Cell adhesion molecules have long been implicated in the regulation of axon growth, but the precise cellular roles played by individual cell adhesion molecules and the molecular basis for their action are still not well understood. We have used the sensory system of the Drosophila embryo to shed light on the mechanism by which the L1-type cell adhesion molecule Neuroglian regulates axon growth. RESULTS: We have found a highly penetrant sensory axon stalling phenotype in neuroglian mutant embryos. Axons stalled at a variety of positions along their normal trajectory, but most commonly in the periphery some distance along the peripheral nerve. All lateral and dorsal cluster sensory neurons examined, except for the dorsal cluster neuron dbd, showed stalling. Sensory axons were never seen to project along inappropriate pathways in neuroglian mutants and stalled axons showed normal patterns of fasciculation within nerves. The growth cones of stalled axons possessed a simple morphology, similar to their appearance in wild type embryos when advancing along nerves. Driving expression of the wild-type form of Neuroglian in sensory neurons alone rescued the neuroglian mutant phenotype of both pioneering and follower neurons. A partial rescue was achieved by expressing the Neuroglian extracellular domain. Over/mis-expression of Neuroglian in all neurons, oenocytes or trachea had no apparent effect on sensory axon growth. CONCLUSION: We conclude that Neuroglian is necessary to maintain axon advance along axonal substrates, but is not required for initiation of axon outgrowth, axon fasciculation or recognition of correct growth substrates. Expression of Neuroglian in sensory neurons alone is sufficient to promote axon advance and the intracellular region of the molecule is largely dispensable for this function. It is unlikely, therefore, that Nrg acts as a molecular 'clutch' to couple adhesion of F-actin within the growth cone to the extracellular substrate. Rather, we suggest that Neuroglian mediates sensory axon advance by promoting adhesion of the surface of the growth cone to its substrate. Our finding that stalling of a pioneer sensory neuron is rescued by driving Neuroglian in sensory neurons alone may suggest that Neuroglian can act in a heterophilic fashion. PMID- 18397532 TI - Molecular signature of hypersaline adaptation: insights from genome and proteome composition of halophilic prokaryotes. AB - BACKGROUND: Halophilic prokaryotes are adapted to thrive in extreme conditions of salinity. Identification and analysis of distinct macromolecular characteristics of halophiles provide insight into the factors responsible for their adaptation to high-salt environments. The current report presents an extensive and systematic comparative analysis of genome and proteome composition of halophilic and non-halophilic microorganisms, with a view to identify such macromolecular signatures of haloadaptation. RESULTS: Comparative analysis of the genomes and proteomes of halophiles and non-halophiles reveals some common trends in halophiles that transcend the boundary of phylogenetic relationship and the genomic GC-content of the species. At the protein level, halophilic species are characterized by low hydrophobicity, over-representation of acidic residues, especially Asp, under-representation of Cys, lower propensities for helix formation and higher propensities for coil structure. At the DNA level, the dinucleotide abundance profiles of halophilic genomes bear some common characteristics, which are quite distinct from those of non-halophiles, and hence may be regarded as specific genomic signatures for salt-adaptation. The synonymous codon usage in halophiles also exhibits similar patterns regardless of their long-term evolutionary history. CONCLUSION: The generality of molecular signatures for environmental adaptation of extreme salt-loving organisms, demonstrated in the present study, advocates the convergent evolution of halophilic species towards specific genome and amino acid composition, irrespective of their varying GC-bias and widely disparate taxonomic positions. The adapted features of halophiles seem to be related to physical principles governing DNA and protein stability, in response to the extreme environmental conditions under which they thrive. PMID- 18397537 TI - Nasal dryness discomfort in individuals receiving dry oxygen via nasal cannula. PMID- 18397538 TI - Vocalization during huff. PMID- 18397539 TI - Obstacles to implementing evidence-based guidelines. PMID- 18397542 TI - Hypoxia in adipose tissue: a basis for the dysregulation of tissue function in obesity? AB - White adipose tissue is a key endocrine and secretory organ, releasing multiple adipokines, many of which are linked to inflammation and immunity. During the expansion of adipose tissue mass in obesity there is a major inflammatory response in the tissue with increased expression and release of inflammation related adipokines, including IL-6, leptin, monocyte chemoattractant protein-1 and TNF-alpha, together with decreased adiponectin production. We proposed in 2004 (Trayhurn & Wood, Br J Nutr 92, 347-355) that inflammation in adipose tissue in obesity is a response to hypoxia in enlarged adipocytes distant from the vasculature. Hypoxia has now been directly demonstrated in adipose tissue of several obese mouse models (ob/ob, KKAy, diet-induced) and molecular studies indicate that the level of the hypoxia-inducible transcription factor, hypoxia inducible factor-1 alpha, is increased, as is expression of the hypoxia-sensitive marker gene, GLUT1. Cell- culture studies on murine and human adipocytes show that hypoxia (induced by low O2 or chemically) leads to stimulation of the expression and secretion of a number of inflammation-related adipokines, including angiopoietin-like protein 4, IL-6, leptin, macrophage migration inhibitory factor and vascular endothelial growth factor. Hypoxia also stimulates the inflammatory response of macrophages and inhibits adipocyte differentiation from preadipocytes. GLUT1 gene expression, protein level and glucose transport by human adipocytes are markedly increased by hypoxia, indicating that low O2 tension stimulates glucose utilisation. It is suggested that hypoxia has a pervasive effect on adipocyte metabolism and on overall adipose tissue function, underpinning the inflammatory response in the tissue in obesity and the subsequent development of obesity-associated diseases, particularly type 2 diabetes and the metabolic syndrome. PMID- 18397543 TI - Technique of high-frequency endolaryngeal ultrasound. AB - OBJECTIVE: We describe in detail the technique of in vivo endolaryngeal ultrasound during microlaryngoscopy, offering a number of practical suggestions which may be of use, especially to the otolaryngologist. METHODS: Endosonography is a non-invasive imaging technology usually applied for the assessment of local tumour growth and regional lymph node metastases in the digestive tract. CONCLUSIONS: Due to a penetrating depth of up to 25 mm and a high tissue resolution, endosonography also qualifies for use in the larynx. The technique may have a potentially important role in the intra- and post-operative investigation of laryngeal lesions bigger than 3 mm. PMID- 18397544 TI - Vascular depression: myth or reality? PMID- 18397545 TI - Wegener's granulomatosis of the orbit: two cases requiring endoscopic surgical decompression. AB - OBJECTIVES: We report two unusual and contrasting cases of Wegener's granulomatosis involving the orbit, requiring urgent endoscopic orbital decompression surgery. CASE SERIES: Both patients presented with an acute deterioration in visual function which failed to improve with medical management. Case one presented with systemic features of the condition, in contrast with case two, who presented with a more localised form of the disease. Whereas case one exhibited primary orbital disease with maxillary sinus extension (i.e. focal disease), case two illustrates orbital involvement secondary to sinus disease extension (i.e. contiguous disease). Prompt diagnosis, assisted by the presence of systemic features, led to a good visual outcome in case one. In case two, in which diagnosis was difficult and surgery delayed, the outcome was poor. CONCLUSIONS: These cases of orbital Wegener's granulomatosis illustrate the diagnostic challenge, the requirement for early intervention following acute visual deterioration and the importance of heightened awareness of the rarer ENT manifestations of this disease. PMID- 18397546 TI - Volunteering as a predictor of all-cause mortality: what aspects of volunteering really matter? AB - BACKGROUND: This study evaluates the predictive effects of different aspects of volunteering (e.g. volunteering status, number of hours, number of years, and type of volunteering activity) on all-cause mortality. METHODS: A seven-year follow-up dataset of a nationally representative sample of Israelis, 60 years and older was used. RESULTS: As expected, volunteering was associated with a reduced mortality risk even after adjusting for age, gender, education, baseline mental health and physical health, activity level, and social engagement. Those who volunteered for 10 to 14 years had a reduced mortality risk relative to non volunteers. In addition, those who volunteered privately, not as part of an official organization, also had a reduced mortality risk compared to non volunteers. The number of hours of volunteering was not a significant predictor of all-cause mortality in the fully adjusted model. In additional sensitivity analyses limited to those who volunteered, none of the various aspects of volunteering was associated with a reduced mortality risk. CONCLUSIONS: Results suggest that not all aspects of volunteering have the same predictive value and that the protective effects of length of volunteering time and type of volunteering are particularly important. However, whether or not volunteering is the most consistent predictor of mortality and whether once a person volunteers the various aspects of volunteering are no longer associated with mortality risk. PMID- 18397547 TI - The association between depressive and cognitive symptoms in amnestic mild cognitive impairment. AB - BACKGROUND: Depressive symptoms are frequently observed in older adults with mild cognitive impairment (MCI). However, little is known regarding the cognitive characteristics of this important subgroup. METHODS: We examined executive functions (controlled inhibition) and verbal episodic memory in 33 healthy older adults (control group), 18 older adults with amnestic MCI plus subclinical depressive symptoms (a-MCI/D+ group), and 26 older adults with amnestic MCI but no depressive symptoms (a-MCI group). RESULTS: Compared to the a-MCI and control groups, patients with a-MCI/D+ showed poor controlled inhibition. Moreover, in verbal episodic memory these patients recalled fewer words than control participants on immediate free, delayed free, and delayed total (free plus cued) recall. Performance on immediate recall suggested a self-retrieval deficit, but delayed performance also revealed the existence of an encoding impairment. In the a-MCI group, participants exhibited normal performance on the executive task, but pervasive memory impairment; the memory deficit concerned free and total recall on both immediate and delayed tasks, suggesting the existence of encoding and self-retrieval disturbances. CONCLUSIONS: This study reveals differences between the pattern of cognitive impairment for a-MCI/D+ and a-MCI subgroups particularly at the level of executive capacities. In terms of memory functioning, the differences between the subgroups were more subtle; more studies are needed in order to better characterize the memory impairment of a-MCI/D+ and a-MCI patients. PMID- 18397548 TI - Factors associated with persistent risk of depression in older people following discharge from an acute cardiac unit. AB - BACKGROUND: This longitudinal study aims to describe the prevalence and characteristics associated with persistent risk of depression in a group of older, hospitalized patients. METHODS: We examined patients at two time-points: baseline and one month post-discharge from hospital. Patients in this study comprised those who had been admitted to the cardiology unit, with no cognitive impairment, aged 60 years and over, and those who were followed up at both time points (N = 155). Questionnaires administered included risk of depression (Geriatric Depression Scale-15; GDS-15), cognitive impairment (Mini-mental State Examination), social support (7-Item Subjective Social Support Index), co morbidity (Charlson's Comorbidity Index), sociodemographic variables, physical functioning (Modified Barthel's Index) and clinical variables. RESULTS: The prevalence of risk of depression (GDS-15 score > or = 5) among older inpatients at baseline was 34%. At one month post-discharge this had fallen to 17% and this group was identified as those at persistent risk of depression. Factors associated with a risk of persistent depression were: hospitalization within the last six months; length of stay of four days or more; discharge diagnosis of angina; and impaired Subjective Social Support Score. CONCLUSION: Depression occurs commonly among older hospitalized patients and may resolve spontaneously. The identification of factors associated with persistent risk of depression can be helpful when looking at which patients may benefit most from screening and treatment for depression after discharge. PMID- 18397549 TI - Switching from risperidone to olanzapine in a one-year, randomized, open-label effectiveness study of schizophrenia. AB - OBJECTIVE: Switching medications is common in the treatment of schizophrenia. This study examines the effectiveness of olanzapine therapy following a clinically warranted switch from risperidone during treatment of patients with schizophrenia. RESEARCH DESIGN AND METHODS: This post-hoc analysis used data from the risperidone arm of a randomized, open-label, 1-year study of patients with schizophrenia. Study protocol permitted antipsychotic switching when clinically warranted, and outcomes were assessed with standard psychiatric measures. Statistical analyses assessed changes from pre- to post-medication switch and endpoint comparisons between patients switched from risperidone to olanzapine and patients continued on risperidone. RESULTS: Most patients who switched from risperidone switched to olanzapine (43/60; 71.7%). Average duration of risperidone treatment prior to switching was 86 days (mean modal dose 4.0 mg/day). Most switchers (86%) completed the 1-year study on olanzapine (average duration 241 days; mean modal dose 12.0 mg/day). Following switch to olanzapine, patients experienced significant improvements on clinical (Brief Psychiatric Rating Scale) and social (Quality of Life Inventory) parameters, with similar proportions of patients achieving remission status at endpoint compared with risperidone patients not requiring medication switch (41.9 vs. 35.5%). Mean weight gain for switchers was approximately 0.4 kg while on risperidone (average treatment duration < 3 months) and 2.4 kg on olanzapine (average treatment duration approximately 8 months). CONCLUSIONS: This study suggests that olanzapine is an effective treatment option for schizophrenia patients requiring a switch from risperidone. Given the small sample size and lack of a comparative group, one cannot determine if other medication options would have been as effective as the switch to olanzapine. Thus, further research is warranted. PMID- 18397550 TI - Primary syphilis. AB - Cases of syphilis have been increasing in the UK and it remains an important public health problem. Here, we provide an overview of syphilis, its presentation, diagnosis and management. PMID- 18397551 TI - The polymicrobial hypothesis of bacterial vaginosis causation: a reassessment. AB - The cause of bacterial vaginosis remains controversial. The two leading hypotheses are that Gardnerella vaginalis is the specific aetiologic agent versus the polymicrobial hypothesis that G. vaginalis acts in concert with other bacteria, principally anaerobes, to produce the disease. Here we reassess the prevailing polymicrobial hypothesis, finding it conceptually problematical. No host initiating factors as postulated have been identified, nor is there conclusive evidence that G. vaginalis lacks diagnostic specificity for the disease. The polymicrobial hypothesis, presupposing a unique disruption of vaginal microecology, is inconsistent with the epidemiological profile of the syndrome, which is that of a sexually transmitted disease. The epidemiological and clinical similarities between bacterial vaginosis and trichomoniasis suggest a similar pathogenetic process; i.e., primary causation by a specific agent, with secondary anaerobic activation and resultant amine production. PMID- 18397552 TI - Asymptomatic urethral infection in male sexually transmitted disease clinic attendees. AB - The purpose of this study is to determine the prevalence of asymptomatic male patients with urethral infections attending a government sexually transmitted infection clinic in Hong Kong and their microbiological profile. A total of 274 consecutive male patients without any symptoms for urethral infections were recruited. A questionnaire was used to record the symptoms, sexual history and demographics. Further assessment, including urethral smear for Gram stain, gonococcal culture and polymerase chain reaction (PCR) for Chlamydia trachomatis (CT), Mycoplasma genitalium (MG) and Ureaplasma urealyticum (UU) were performed. In 274 asymptomatic patients, 36 patients had non-gonococcal urethritis (NGU) and two patients had positive gonococcal culture. Among the asymptomatic patients with NGU, there were 6 (16.6%), 10 (22.8%) and five (13.9%) patients with positive PCR for CT, UU and MG, respectively. In addition, there were 14 asymptomatic patients with positive PCR for CT but without evidence of NGU. In conclusion, urethral infections were identified in a significant number of asymptomatic male patients and therefore, routine screening for this group is warranted. PMID- 18397553 TI - Low seroprevalence of herpes simplex virus type 2 among pregnant women in Senegal. AB - Herpes simplex virus type 2 (HSV-2) is considered as a major co-factor of both sexual transmission and acquisition of the human immunodeficiency virus (HIV). The HIV epidemic in Senegal is characterized by a remarkable and stable low prevalence. Whether HSV-2 may also constitute a possible co-factor favouring the spreading of HIV epidemic in Senegal is yet unknown. This prompted us to evaluate the HSV-2 seroprevalence in the sentinel population of pregnant women in Senegal. Two hundred and sixty pregnant women attending Roi Baudouin maternity in the capital city Dakar (n = 14; 135) and the antenatal clinic in Kaolack (n = 125), the third city of Senegal, were prospectively recruited between March and August 2003. Fifty-six women (22%) were positive for HSV-2 serology. The prevalence of HSV-2 seropositivity was higher in women living in Dakar (26%) than in those living in Kaolack (16%) (P < 0.01). Only two women from Dakar and two other from Kaolack were found to be HIV-1-infected. Our observations suggest a seemingly low seroprevalence of HSV-2 infection in adult women Senegal, comparable with those usually reported in Western countries. Further, epidemiological surveys are needed to confirm these results in the general population. PMID- 18397554 TI - An educative genitourinary medicine clinic website: value to users and impact on service demand. AB - In 2004, the Newcastle genitourinary medicine clinic launched a new website. To assess its value to users and impact on service demand, we surveyed those attending the clinic with a new episode between September and December 2005 by a questionnaire. One hundred and fifteen of 601 respondents (19%) visited the website. Fifty-nine percent of users were men with those aged between 35 and 44 ranking the highest percentage (31.7%) among those who visited the website. Fourteen (12.5%) would not have attended if they had not visited the website. One hundred and two (88.7%) found it easy to use and 81 (70.4%) looked for specific information of whom 73 (89%) were successful. Web users were more likely to present with an episode not requiring treatment possibly indicating increased demand promoted by our prevention information but less likely to have bacterial vaginosis, anaerobic balanitis and anogenital candidosis (not statistically significant). PMID- 18397555 TI - Social comparison activity in coping with HIV. AB - Taylor and Lobel's (1989) model of social comparison processes under threat predicts that upward affiliation (a social activity) may be useful to improve the skills and to find inspiration in coping with HIV, whereas downward evaluation (a cognitive activity) may assist someone to feel better by comparing oneself to a less fortunate person. Participants with HIV were randomly assigned, using a 3 x 3 factorial design, to read one of nine vignettes about someone whose HIV medical prognosis and psychological adjustment were described as good, poor or unspecified. Participants desired upward affiliation with a target doing well physically, but they wanted to avoid a target doing poorly psychologically. Participants also evaluated their own physical condition as better than a target coping poorly physically with HIV. The results may be useful in understanding the impact of various role models in coping with HIV. PMID- 18397556 TI - The demographical and clinical features of patients reattending a genitourinary medicine clinic and the role of counselling on subsequent incidence of sexually transmitted infections. AB - The aim of this study was to investigate the incidence of reinfection with sexually transmitted infection (STI) and its possible predicting factors among patients reattending a department of genitourinary (GU) medicine. The method included a retrospective study on consecutive patients re-screened for STI in a busy department of GU medicine between January 1996 and April 2005. Cox regression model was used to investigate the association of age, gender, sex group, previous STI, counselling for an STI on the last attendance, number of sexual partners in the previous attendance episode, condom use and the number for partners with whom they had unprotected sex, with STI at current screening. The results show that 302 consecutive patients with more than two attendance episodes were included in the study. Patients were mostly women (92%), younger than 35 years (80%) and heterosexual (98%). At baseline, 70 (23%) patients had STI. The incidence density of STI at the end of follow-up period was 15.69/100 person years. Testing positive for an STI was associated with age younger than 25 years (hazard ratio 2.70 [95% confidence interval 1.70-4.57]; P = 0.000). In conclusion, our study confirmed the improvement in access to GU medicine services, encouragement of patients with an STI for re-screening on regular basis and use of novel STI counselling strategies, especially for younger patients may reduce the incidence of subsequent STI. PMID- 18397557 TI - Influence of antiretroviral treatment on quality of life in seropositive inmates. AB - The aim of the study is to evaluate the influence of antiretroviral treatment on health-related quality of life (HRQOL) of three groups of HIV-positive inmates: those who are taking antiretroviral treatment, those who are not on treatment as it has not yet been indicated, and those who refuse to take treatment even though it has been recommended. A cross-sectional study was conducted on 585 HIV+ inmates in three prisons. The response variable was HRQOL. Independent variables were: sociodemographic variables, psychosocial and drug-related variables. Two multivariate linear regression models were constructed in order to determine the HRQOL, physical health score (PHS) and mental health score (MHS), for each of the three groups identified, using patients who refused treatment as the reference category. Patients who refused therapy had a lower MHS compared with patients in whom treatment was not indicated (P = 0.038). With regard to PHS, patients refusing therapy had a lower score than patients who were not indicated therapy (P = 0.005), and than patients receiving therapy (P = 0.010). PMID- 18397558 TI - HIV-related knowledge and stigma among high school students in Libya. AB - This study aims to measure indicators of HIV-related stigma among students of high schools in the North West of Libya. The results will be part of baseline data and evaluation of the impact of successive interventions. Understanding the behaviour of risk groups in a society, such as young people, is essential in order to draw effective prevention strategies. Behavioural surveillance surveys have been shown to make an important and useful contribution to informing the response to HIV. This study was part of a large Knowledge, Attitude, Beliefs and Practice survey. A self-administrated questionnaire method was used. The response rates were high (83-92%). Despite high level of stigma shown by both boys and girls in the study, 91% of students supported providing free care to HIV infected individuals. The HIV intervention programmes for young people should operate within a comprehensive strategy to combat HIV/AIDS. The stigmatizing and the discriminatory perceptions of HIV infected individuals should be addressed as part of the education campaign. PMID- 18397559 TI - Preferred options for receiving sexual health screening results: a population and patient survey. AB - Current genitourinary medicine patients (202) and potential future patients (542) completed a questionnaire-based survey to determine their preference for obtaining test results, their acceptability of including a named infection on contact slips and to report expectations about the acceptable length of an appointment. Overall, most respondents (78% [n = 582]) felt it unacceptable to be only contacted if their results were positive ('no news is good news'). In the clinic, a majority preferred a contact slip to be coded (68% [n = 137]), in the general public views were balanced. Significantly, more people in the general population expected an appointment to last no longer than 30 min (32% [n = 173] cf. 10% [n = 21], P < 0.001). A clear preference was expressed to receive sexually transmitted infection test results even if they are negative. Telephone and face-to-face contact were most popular with relatively few choosing mobile telephone text messaging or email as their preferred option. PMID- 18397560 TI - Improved injection network ascertainment with supplementary elicitation techniques. AB - Prior research indicates that injection drug users forget substantial proportions of their injection partners when asked to recall them. Such under-reporting hampers both ascertainment of the injection networks that underlie transmission of blood-borne pathogens and contact-tracing efforts for disease control. We report here an evaluation of supplementary elicitation techniques--a set of prompting strategies and recall cues--employed in a study that involved tracing of injection partners. Sixty-one index drug injectors in Seattle participated in the study. The supplementary partner elicitation techniques enhanced recall of injection partners substantially and identified persons relevant to transmission of blood-borne pathogens. As a set, the supplementary techniques elicited additional partners from 70% of injectors, and the additional partners elicited represent a 75% increase on average. Drug injectors who recalled many partners before administration of the supplementary techniques tended to report more additional partners in response to the supplementary techniques than injectors who freely recalled few partners. In addition, partners elicited by the supplementary techniques were as likely as freely recalled partners to test positive for hepatitis C virus antibody and engage in risk behaviour with indexes. Furthermore, we found that the supplementary techniques boosted connectivity in the observed injection network considerably. PMID- 18397561 TI - Prevalence and factors associated with anal lesions mediated by human papillomavirus in men with HIV/AIDS. AB - Prevalence and risk factors for human papillomavirus-related anal lesions were evaluated in 60 men with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). Patients underwent anal cytology, anuscopy under colposcopic vision and anal biopsy for detection of the lesions. The mean age was 41.9 years and the mean time of HIV infection was 6.8 years, 88.3% of them having been on highly active antiretroviral therapy for an average of 6.5 years. Homosexuals represented 43.3% and bisexuals 15.0% whereas heterosexuals comprised 41.7%. The prevalence of anal lesions detected by anuscopy under colposcopic vision, cytology and biopsy were, respectively, 35.0, 16.7 and 23.3%. Homosexuals or bisexuals accounted for 85.7% of the patients with an abnormal biopsy, the remaining 14.3% being heterosexuals (P = 0.02). The T-CD(4)(+) lymphocyte count, HIV viral load and use of antiretrovirals did not reveal any association with anal lesions. The occurrence of anal lesions was high in the individuals with HIV/AIDS, especially in the homosexuals and bisexuals, but it also occurred in heterosexuals, justifying the screening of anal lesions of all men with the infection. PMID- 18397562 TI - Occult hepatitis B virus infection in HIV-infected Lebanese patients with isolated antibodies to hepatitis B core antigen. AB - The presence of hepatitis B virus (HBV) serological markers have been investigated in 101 Lebanese patients (69 men, 32 women; mean age 32.7 +/- 1.7 years) infected with human immunodeficiency virus type 1 (HIV-1). Seven patients (6.9%) were HBsAg carriers compared with 54 patients (53.5%) who had no evidence of exposure to HBV infection. Twenty-four patients (23.8%) had anti-HBc alone as a serological marker compared with four patients who were positive for anti-HBs alone and 12 patients (11.9%) who were anti-HBc and anti-HBs-positive. Occult HBV infection (presence of HBV DNA in the absence of HBsAg) is found to be relatively high (28.7%) in HIV-infected Lebanese patients and the overwhelming majority (83.3%) of those who were positive for anti-HBc alone had a detectable HBV DNA in their serum. However, none of our HIV-positive patients with occult HBV infection had abnormal alanine aminotransferase level, which also raises the question as to whether occult HBV plays a role in the aetiology of liver disease in HIV-infected patients. Further, studies on the association between HBV DNA levels and markers of liver function in addition to data on liver biopsy would help in answering this question. PMID- 18397563 TI - Testing rates for sexually transmitted infections among HIV-infected men who have sex with men attending two different HIV services. AB - This study compares the testing rates of bacterial sexually transmitted infections (STIs) among HIV-positive men who have sex with men (MSM) attending two HIV clinics in Melbourne. Data on STI testing over a 12-month period were obtained for all HIV-positive MSM who attended the clinics between January and March 2006. Screening rates for bacterial STIs were significantly higher at a sexual health clinic (n = 254) compared with an infectious diseases clinic (n = 351), whether this was measured according to: at least one STI test being performed for chlamydia, gonorrhoea or syphilis (69% vs. 38%, P < 0.01); serological testing for syphilis alone (67% vs. 34%, P < 0.01); or 'complete' STI screening for pharyngeal gonorrhoea, urethral chlamydia, anal gonorrhoea, anal chlamydia and syphilis (41% vs. 6%, P < 0.01). Substantial differences in STI testing rates among HIV-positive MSM may exist between HIV clinical services depending on the measures in place that promote STI screening. PMID- 18397564 TI - Does using self-completed sexual history questionnaires in HIV-positive men who have sex with men affect clinical outcomes? AB - This audit compared sexual histories taken by self-completed questionnaires (SQ) versus the 'gold-standard' of 'traditional' face-to-face interviews by health care professionals (HP). It compared reporting of symptoms and sexual behaviour and looked at outcomes in terms of diagnoses of sexually transmitted infections. SQ were at least equal to HP and might therefore be a valuable tool in streamlining services. PMID- 18397565 TI - Are women from high-risk ethnic minority groups more likely to decline antenatal HIV screening? AB - Eight new cases of human immunodeficiency virus (HIV) were diagnosed in the antenatal population of Milton Keynes within the first two years of our 'opt-out' antenatal testing scheme; the majority (6/8) occurred in women of black African origin. Since it is suggested that individuals from high-risk groups are more likely to decline HIV testing, we were concerned that women from this high-risk ethnic group might not be accepting testing. Such a situation would increase the risk of undiagnosed HIV-positive women delivering at Milton Keynes and undermine the potential benefits of the screening programme. Retrospective review of pregnant women delivering in our area over six months was performed. Hospital obstetric and microbiology databases were analysed for results of HIV screening and ethnic origin of patients. A total of 1586 women delivered during the study period. Among the black African women 13/158 (8.2%) declined screening, compared with 120/1214 (9.8%) and 15/153 (9.8%) of white and Asian women, respectively. The high uptake of testing across all groups suggests that the policy of offering and recommending HIV screening to all women is being appropriately implemented. Black African women were more likely to have undergone screening than white or Asian women, although the differences were not statistically significant. PMID- 18397566 TI - Fulminant autoimmune hepatitis after successful interferon treatment in an HIV HCV co-infected patient. AB - Hepatitis C (HCV) treatment using interferon-alpha (IFN-alpha) and ribavirin is recommended in HIV/HCV co-infected patients to prevent liver cirrhosis and liver related death. However, in addition to its antiviral activity, IFN is a pleiotropic cytokine able to synergistically amplify T-cell autoreactivity. Here, we report for the first time the induction of a subfulminant autoimmune hepatitis (AIH) after four months of a successful treatment of HCV-1b infection using peg IFN and ribavirin, in a 48-year-old woman co-infected with HIV. Diagnosis was assessed according to the international AIH scoring system, including liver biopsy and confirmed by positive response to steroid challenge. PMID- 18397567 TI - Pregnancy-induced lymphangiectasias of the vulva. AB - Lymphangiectasias occurring as a consequence of pregnancy and spontaneously regressing upon successful outcome of pregnancy is a rarity that needs to be reported. More so because these lesions occurring on the vulva can mimic various other entities and can sometimes be misconstrued as sexually transmitted disease by persons involved in primary care. This is a case report of lymphangiectasias occurring during pregnancy. PMID- 18397568 TI - Be wary, this is not a case of vulval warts! AB - We present a 40-year-old woman who presented to the genitourinary medicine department with a two-year history of recurrent non-painful vulval lumps. She was in a stable relation with one male sexual partner over the preceding 20 years and her sexual health screen was negative. Clinical examination of these lumps was suggestive of anogenital warts for which she has been treated on numerous occasions over the last two-year-period with liquid nitrogen cryotherapy and podophyllotoxin 0.15% cream. As the lumps were not responding to conventional wart treatment, a biopsy from these lesions was performed. The histological appearance showed focal fibrin thrombus formation within dilated vascular channels consistent of a subepidermal cavernous haemangioma. This case demonstrates that the clinical diagnosis of anogenital warts is not always straightforward. It highlights the importance of obtaining a tissue diagnosis from the resistant or atypically looking warty lesions to avoid making the wrong diagnosis. PMID- 18397569 TI - Syphilis and disadvantage in rural communities. PMID- 18397570 TI - Answers to CPD test questions. PMID- 18397571 TI - In vivo versus in vitro airway surface liquid nicotine levels following cigarette smoke exposure. AB - Whole cigarette smoke (WCS) is composed of approximately 5% particulates and 95% vapors by weight and is difficult to reproduce quantitatively in the laboratory, where typically, routine in vitro application of smoke normally only utilizes the particulate phase. In this study, we used a system for exposing epithelial cells cultured at an air-liquid interface to WCS. We hypothesized that the use of WSC in vitro was more relevant to what is seen in vivo than methods of cigarette smoke application that only use a small fraction of WCS [i.e., aqueous extract or cigarette smoke condensate (CSC)]. To test this hypothesis, we compared nicotine and cotinine concentrations (measured by mass spectrometry) in the airway surface liquid (ASL) of human primary bronchial epithelial cultures (HBECs) exposed to serial dilutions of WCS to the concentrations found in induced sputum of human subjects who had recently smoked a cigarette; this was also compared to the concentrations found after an exposure to a concentration of CSC commonly used in vitro. When measured by mass spectrometry, nicotine levels were not significantly different in induced sputum versus the ASL of HBECs exposed in vitro to a 1:30 exposure of WCS. However, HBECs that had been exposed to CSC returned significantly lower concentrations of ASL nicotine. These results suggest that nicotine is a good dosimetry marker of WCS exposure and provides direct evidence that the use of WCS is more relevant than the use of CSC for in vitro systems. PMID- 18397572 TI - An enantiomer-selective liquid chromatography-tandem mass spectrometry method for methadone and EDDP validated for use in human plasma, urine, and liver microsomes. AB - A liquid chromatography-electrospray ionization-tandem mass spectrometry method has been developed and validated to detect (R)- and (S)-methadone and (R)- and (S)-2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in human plasma with cross-validation to urine and liver microsomes. Use of deuterated internal standards and liquid-liquid extraction coupled with chiral separation provided baseline separation with a lower limit of quantitation (LLOQ) of 2.5 ng/mL. The LLOQ was established from comparison of signal in blanks from six different sources per matrix with the same sources fortified at the LLOQ (none exceeded 19% of LLOQ) and precision and accuracy at the LLOQ determined in the same six sources per matrix. The assay was precise (% coefficients of variation within 13.8%) and accurate (% targets within 15%) in all three matrices. No interference was seen from addition of other psychoactive drugs. Stability was determined in plasma (24 h at room temperature, 321 days at -20 degrees C, 3 freeze-thaw cycles); processed plasma samples (5 days at -20 degrees C, 12 days on autosampler); urine (24 h at room temperature); and stock solutions (20 h at room temperature, 61 days at -20 degrees C). Applications of varying degree are presented for each matrix. Plasma from five subjects maintained on 100 mg oral methadone per day permitted comparison of the pharmacokinetics of the enantiomers. The t(1/2) of (R)-methadone was significantly longer than for (S) methadone, and (S)-methadone was more tightly protein bound. The C(max), AUC, C(min), and % protein bound of (S)-EDDP were significantly greater than (R)-EDDP, while the t(1/2) of (R)-EDDP was significantly greater than (S)-EDDP. In spot urines, (R)- was higher than (S)-methadone, and (S)- was generally higher than (R)-EDDP. (R)- and (S)-EDDP production was detected after incubation of therapeutic concentrations of racemic methadone with human liver microsomes, and (S)-EDDP production was twofold greater than (R)-EDDP in three human placental microsomes incubated with racemic methadone. PMID- 18397573 TI - Development and validation of ELISA and GC-MS procedures for the quantification of dextromethorphan and its main metabolite dextrorphan in urine and oral fluid. AB - The development of a highly sensitive enzyme-linked immunosorbent assay and gas chromatography-mass spectrometry confirmation method for the detection of dextromethorphan and its major metabolite dextrorphan in urine and oral fluid is described. For the screening assay, the intraday precision was less than 8% for urine and less than 5% for oral fluid. The interday precision was less than 10% for both drugs in urine and oral fluid. For the confirmatory procedure, both inter- and intraday precision was less than 5% for both matrices. The detection limit for both methods was 1 ng/mL. The quantifying ions chosen from the full scan mass spectra were m/z 271 for dextromethorphan, m/z 329 for dextrorphan, and m/z 332 for tri-deuterated dextrorphan-d(3). A high recovery yield (> 93%) from the Quantisal oral fluid collection device was achieved, and the drugs were stable in the collection device for at least 10 days at room temperature. The extracted drugs from both matrices were stable for at least 48 h while kept at room temperature. Both screening and confirmatory procedures were applied to authentic urine and oral fluid specimens obtained from volunteers following therapeutic ingestion of dextromethorphan. PMID- 18397574 TI - Development and clinical application of an LC-MS-MS method for mescaline in urine. AB - Mescaline (3,4,5-trimethoxyphenylethylamine) is an hallucinogenic psychoactive substance present in several species of cacti. Mescaline has a documented use dating back 5700 years. In more recent years, the interest in hallucinogenic designer drugs such as ecstasy has also triggered interest in the naturally occurring mescaline. This study was undertaken to develop a liquid chromatography tandem mass spectrometry (LC-MS-MS) method for the screening and confirmation of mescaline in human urine samples and to apply this method to routine testing in patient samples. For the screening procedure, chromatographic separation was achieved on a 5-microm HyPURITY C(18) column, using a methanol gradient in ammonium acetate buffer. The MS-MS analysis was performed using selected reaction monitoring; the transitions monitored were m/z 212.3 --> m/z 180.3 for mescaline and m/z 221.3 --> m/z 186.3 for the deuterated internal standard (mescaline d(9)). The detection limit for mescaline in urine matrix was 3-5 microg/L, the upper limit of quantification was 10,000 microg/L, and the total coefficient of variation for spiked samples containing 10 to 1025 microg/L was < 8.5%. The confirmation procedure included a sample clean-up by solid-phase extraction on a C(18) cartridge, and one extra transition for mescaline (m/z 212.3 --> m/z 195.2) was monitored. The LC-MS-MS method was found to be sensitive and specific for the routine detection of mescaline in urine. Among 462 urine samples collected from young people with alcohol or drug problems, 32% were positive for illicit drugs, but none for mescaline. PMID- 18397575 TI - Analysis of confiscated black market drugs using chromatographic and mass spectrometric approaches. AB - In the context of house searches in Germany, numerous drugs were confiscated and subjected to chemical analysis, including anabolic agents such as various anabolic-androgenic steroids (stanozolol, testosterone derivatives, trenbolone esters, etc.) and clenbuterol, as well as agents with anti-estrogenic activity (tamoxifen, clomiphene), drugs stimulating virility (sildenafil, tadalafil), and unlabeled plastic bags. Liquid chromatography-tandem mass spectrometry, gas chromatography-mass spectrometry with nitrogen-phosphorus specific detection, gel electrophoresis, and immunological tests were employed to test for the effective content of 70 products. In 18 cases (25.7%), the declared ingredients differed from the actual content, in particular concerning anabolic-androgenic steroids. Nandrolone and trenbolone esters, for instance, were frequently substituted or complemented by various testosterone derivatives, and several testosterone depot formulations originally composed of four different esters were found to contain fewer or wrong components. Except for those drugs supposedly originating from so called underground labs, fake packings were hardly or not distinguishable from original boxes by visual inspection. PMID- 18397576 TI - Formic acid and methanol concentrations in death investigations. AB - Methanol ingestion results in the formation of formic acid, a toxic metabolite that can cause metabolic acidosis. Methanol toxicity is therefore dependent on the amount of methanol ingested, the nature of treatment received, elapsed time since ingestion, and the accumulation of formic acid. Both methanol and formic acid concentrations are determined at this laboratory using headspace gas chromatography. An examination of 12 fatalities attributed to methanol poisoning is presented. Six individuals were found deceased, and their postmortem methanol and formic acid concentrations ranged from 84 to 543 mg/dL and 64 to 110 mg/dL, respectively. In the other six individuals, hospital treatment such as bicarbonate, ethanol infusion, and hemodialysis was administered. Antemortem methanol and formic acid concentrations ranged from 68 to 427 mg/dL and 37 to 91 mg/dL, respectively, whereas corresponding postmortem methanol and formic acid levels ranged from undetectable to 49 mg/dL and undetectable to 48 mg/dL, respectively. Hospital treatment of formic acid toxicity resulted in significantly reduced postmortem methanol and formic acid concentrations. Furthermore, the toxicological relevance of nine methanol-positive cases where postmortem methanol concentrations ranged from 3 to 142 mg/dL, with corresponding formic acid levels of less than 10 mg/dL, is discussed. PMID- 18397577 TI - Determination of lovastatin hydroxy acid in female B6C3F1 mouse serum. AB - A liquid chromatographic-mass spectrometricmethod for the determination of lovastatin hydroxy acid in female B6C3F(1) mouse serum was developed for use in supporting toxicokinetic studies of animals dosed with the cholesterol lowering agent lovastatin. The method does not require an extensive sample cleanup and shows good correlation between serum matrix standards and solvent standards. The method was validated and used to analyze serum samples from a preliminary dose level range-finding study. The method was validated for a concentration range of approximatel 1.0 to 100 ng/mL in serum, and linearity was verified to ~2000 ng/mL. The stability of sample extracts was determined under various storage conditions and the stability of serum samples stored frozen was determined over a period of seven weeks. During the course of analyzing the animal samples, the serum was monitored for the presence of lovastatin not hydrolyzed to the hydroxy acid, but no attempt was made to quantify lovastatin. No unhydrolyzed lovastatin was noted in any of the serum samples from animals dosed with lovastatin. PMID- 18397578 TI - Simultaneous determination of ketamine, tramadol, methadone, and their metabolites in urine by gas chromatography-mass spectrometry. AB - An automated solid-phase extraction procedure combined with gas chromatography mass spectrometry methodology, without derivatization, has been developed for the identification and quantitation of ketamine, norketamine, tramadol, methadone, 2 ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine, and 2-ethyl-5-methyl-3,3 diphenylpyrroline in urine. The analytical approach is simple and rapid, yet reliable. Good linearity (r(2) > 0.995 over the concentration range of 30 to 600 ng/mL), sensitivity (limits of quantitation 15-30 ng/mL), accuracy (81.0-109.9%), precision (RSD < 13.8%), and recovery (> 79.6% in average) were achieved for all analytes. Ninety-one urine specimens from suspected drug users and 21 clinical urine specimens from methadone substitution therapy patients were analyzed to validate the method compatibility and stability. Results have demonstrated that this GC-MS method is a good confirmation and quantitation test scheme for the six target compounds in urine. PMID- 18397579 TI - Determination of fentanyl in sweat and hair of a patient using transdermal patches. AB - Fentanyl is a synthetic opioid used to treat intense chronic pain. In this study, the authors report detection and quantification of fentanyl in sweat and hair of a patient receiving fentanyl (25 microg/h) via a transdermal therapeutic system (TTS) for 22 days. Sweat was collected using sweat patches every night on days 13 21 of the therapy, and hair was collected 12 weeks after the end of the treatment. Detection and quantification was performed with liquid chromatography tandem mass spectrometry using electron spray ionization in selected reaction monitoring mode. Alfentanyl was used as internal standard for quantification in hair and in sweat. Sodium ions have been used as endogenous internal reference for determination of volume of sweat excreted on each patch. Results show presence of fentanyl in both matrices. Fentanyl concentrations in sweat varied from 0.17 to 1.02 ng/microL, and time-resolved segmented hair analysis showed a maximum fentanyl concentration of 0.48 ng/mg of hair during the period of the therapy. PMID- 18397580 TI - [Changes in the social networks of older adults who are beneficiaries of public housing projects in Chile]. AB - OBJECTIVES: To identify, compare, and analyze the social networks of two groups of poor, older adults in two types of public housing projects in Chile, in terms of the types of relationships and the frequency and purpose of their interactions, before and after moving to their new homes. METHODS: A descriptive study was conducted of public housing beneficiaries in apartment buildings versus condominiums. The study pool was the group of people assigned to housing in buildings (n = 152) and condominiums (n = 124) from 1998 to 2001. A questionnaire was administered in their homes. RESULTS: Most study participants indicated that the move to public housing meant leaving their old neighborhood for a new one, a change that, among other consequences, produced changes in their social networks. In general, both groups have similar social networks, with their children being the strongest source of support, both before and after the move. After the move, social interaction generally decreased, except among the adults living in apartment buildings, for whom it increased slightly with family members, and in a superficial manner, with others who had weak connections and were neither friends nor family. Interactions with establishments also decreased, although interactions with senior services increased for the condo residents from 28% to 65% and for apartment residents from 31% to 45%. CONCLUSIONS: It is recommended that similar programs strive to preserve and strengthen the social networks in place prior to moving and encourage the formation of relationships in the new residences. PMID- 18397581 TI - Health-related quality of life in a binational population with diabetes at the Texas-Mexico border. AB - OBJECTIVES: To examine physical and mental health domains of health-related quality of life (HRQL) in a binational adult population with type 2 diabetes at the Texas-Mexico border, and to explore individual and social correlates to physical and mental health status. METHODS: Adults 18 years and older with type 2 diabetes residing in the South Texas Lower Rio Grande Valley and in Reynosa, Tamaulipas, Mexico, were recruited using a convenience sampling technique and interviewed face-to-face with a structured survey. HRQL was measured using physical and mental health summary components of the Medical Outcomes Study Short Form. HRQL correlates included demographic characteristics, health factors, access to healthcare, and family support. Samples characteristics were compared using the Student's t-test or Mann-Whitney U test. Associations between dependent and independent variables were examined using unadjusted and adjusted (multiple variable) logistic regression models. RESULTS: There were no significant differences between Valley and Reynosa respondents in physical or mental health status scores. Valley participants with lower socioeconomic status and those perceiving their supportive relative's level of diabetes-related knowledge as "low" were more likely to report worse physical health than those lacking those characteristics. In the Reynosa group, lower physical health status was associated with duration of diabetes and insulin use. Both sample populations with clinical depressive symptoms were more likely to have worse physical and mental health than those without such symptoms. CONCLUSIONS: HRQL is an important outcome in monitoring health status. Understanding the levels and influences of HRQL in U.S.-Mexico border residents with diabetes may help improve diabetes management programs. PMID- 18397582 TI - Seroprevalence and risk factors of Toxoplasma gondii infection among pregnant women in Trinidad and Tobago. AB - OBJECTIVE: To determine the seroprevalence of toxoplasmosis and the frequency of risk factors for the disease in women attending antenatal clinics in hospitals and local health centers in Trinidad and Tobago. METHODS: From November 2002 September 2003, 232 pregnant women at the antenatal clinics of two large hospitals were sampled during their first trimesters. From October 2003-February 2005, 218 pregnant women at five health care centers were followed through three trimesters, wherever possible, and blood samples collected. Sera were screened for Toxoplasma gondii IgG and IgM immunoglobulins using an enzyme immunoassay. Data on demographics, as well as practices considered to be risk factors for toxoplasmosis, were obtained. RESULTS: Of the 232 women sampled at the two hospital clinics, 83 (35.8%) and 8 (3.4%) were seropositive for immunoglobulins of T. gondii IgG and IgM, respectively. Of the 218 women at the health centers, 76 (34.9%) had evidence of past infection (IgG), while 26 (11.9%) were seropositive for IgM immunoglobulin, suggesting new infections during pregnancy. Only having "3 or more" children was significantly associated with infection by T. gondii. CONCLUSION: In Trinidad and Tobago, the seroprevalence of past infection by T. gondii in pregnant women is relatively low (39.3%) for patients in both hospital and health center clinics. Consequently, there is an elevated risk of primary infection during pregnancy and the potential for congenital infection. PMID- 18397583 TI - [Physical activity among children and adolescents in a rural community in Mozambique: effects of age, sex, and nutritional status]. AB - OBJECTIVE: To determine the association between physical activity levels of children and adolescents of a rural community in Mozambique and their age, sex, and nutritional status. METHOD: The sample comprised 845 boys and girls from 7-16 years of age. Weight and height were measured, while physical activity level was determined using a questionnaire developed and validated for the population of Mozambique. Nutritional status was determined according to criteria recommended by the World Health Organization. Analysis of variance (ANOVA) and analysis of covariance (ANCOVA) were used to test the interaction between age and sex and between nutritional groups, respectively. RESULTS: Age and sex significantly influenced all types of activities except walking, for which the effect of sex was not significant (F = 0.4, P = 0.517). Girls were more active than boys. Age and sex were significantly associated only in relation to household chores (F = 1.933, P = 0.044) and sports (F = 2.32, P =0.014). In boys, only the total activity coefficient was different among nutritional groups (F = 3.44, P = 0.017), with boys of normal weight presenting a higher level of activity as compared to boys with low height or both low weight and height. In girls, significant differences were observed only in relation to household chores (F = 2.65, P = 0.049), with girls of normal and low weight differing from the other nutritional groups and those with low height differing from girls low in both weight and height. CONCLUSIONS: The results suggest that in rural Mozambique, deficits in height, weight, or both did not limit the physical activity levels of children and adolescents. PMID- 18397584 TI - [Prescription patterns for antilipidemic drugs in a group of Colombian patients]. AB - OBJECTIVES: To determine patterns in antilipidemic drug prescriptions among a group of patients covered by the General Social Security System (Sistema General de Seguridad Social) in Colombia. METHODS: A descriptive, observational study was conducted of 41 580 hyperlipidemics of both sexes, who were over 20 years of age, undergoing treatment from at least April to June 2006, and were residents of one of 19 cities in Colombia. A database was created to track prescription data collected by the pharmaceutical company that dispenses medications to the patients. RESULTS: The mean age was 58.4+/-13.5 years; 58.9% of the participants were women. Of the total number of patients, 95.6% were receiving monotherapy, while 4.4% were receiving two or more antilipidemics. Prescriptions were ranked as follows: statins (70.9%), fibrates (27.5%), bile acid sequestrant resins (0.9%), and others (0.7%), all at low dosage levels. The most common therapy combinations were lovastatin + gemfibrozil (n = 1 568), cholestyramine + gemfibrozil (n = 92), and cholestyramine + lovastatin (n = 78). Comedications most frequently prescribed were: antihypertensive (60.9%), antiinflammatory (56.5%), antiulcer (22.9%), and antidiabetes drugs (20.6%), and acetylsalicylic acid (ASA, 3.8%). Antianginals and ASA were being underused, while antiinflamatories and antiulcer drugs were being overused. CONCLUSIONS: Dyslipidemia is a primary risk factor for developing coronary heart disease and stroke, frequent causes of morbidity and mortality in Colombia and the world. All of the antilipidemics are being used at lower-than-recommended dosage levels. Clearly there is a need for creating educational strategies to address these prescribing habits and for exploring clinical results of the pharmaceuticals studied. PMID- 18397585 TI - The meaning and consequences of tuberculosis for an at-risk urban group in Ecuador. AB - OBJECTIVE: To explore knowledge, beliefs, perceptions, and attitudes about tuberculosis (TB) in a high-risk group in Ecuador. This included signs and symptoms, causation, transmission, treatment, treatment adherence, impact on lifestyle and role functioning, and stigma. METHODS: A convenience sample of 212 adults undergoing diagnostic TB testing at a public health facility in Quito, Ecuador, was recruited for the study. Data were collected from subjects during face-to-face interviews using a structured instrument containing closed and openended questions. Descriptive and bivariate statistics were used for quantitative analyses; content analysis was used to analyze qualitative data. RESULTS: Most subjects were familiar with TB and some of its characteristics and treatment aspects. However, many also held misconceptions or lacked key knowledge which could adversely affect early diagnosis and treatment and adherence to treatment, and thereby allow the disease to spread. Subject education was the single most important predictor of knowledge, beliefs, perceptions, and attitudes followed by gender, age, and prior disease experience. The subjects linked TB to multiple adverse health, economic, psychological, and social consequences, including stigma. Although none knew if they had TB when interviewed, many reported feeling stigmatized just by being tested. The subjects identified a strong need for formal educational opportunities to learn about TB prevention and control but had little access to these. CONCLUSIONS: The study findings highlight a need for enhanced population access to TB education. Health education and social marketing directed toward increasing TB knowledge and changing perceptions and attitudes could ultimately contribute to improved early diagnosis, treatment adherence, prevention, and decreased stigma. This could be accomplished providing that the public health infrastructure is adequate to meet demands. PMID- 18397586 TI - [Therapeutic uses of investigational drugs: research extension, compassionate use, and expanded access]. AB - This article describes the methodological, regulatory, and ethical aspects of the different therapeutic uses of investigational drugs--research extension, compassionate use, and expanded access. Worldwide, the principle challenges of this kind of treatment are: setting minimum quality standards for researchers, as well as institutions, so that projects can include drugs at various stages of development; training of evaluation and assessment committees on the methodological, regulatory, and ethical aspects of new drug research; clearly outlining the relationship between researchers and funding organizations and between researchers and study participants; and understanding the opposition to the recent proposal to enable drug manufacturers to charge for drugs used in research studies. PMID- 18397587 TI - Making secondary care a primary concern: the rural hospital in Ecuador. PMID- 18397588 TI - [Ibero-American Program of Science and Technology for Development]. PMID- 18397589 TI - Don't believe everything that you read. PMID- 18397590 TI - Research quality and integrity. PMID- 18397591 TI - Unravelling referral paths relating to the dental care of children: a study in Liverpool. AB - OBJECTIVE: To describe primary care referral networks relating to children's dental care and the main influences on referral decisions taken by dentists working in a primary care setting. DESIGN: A postal questionnaire to all 130 general dental practitioners (GDPs) in contract with Primary Care Trusts (PCTs), and 24 Community Dental Service (CDS) dentists in Liverpool. OUTCOME MEASURES: Characteristics of patient groups and factors influencing the choice of referral pathway of children referred from primary dental care. RESULTS: There were good responses rates (110 [85%] GDPs and 22 [92%] CDS dentists). The two main reasons why GDPs referred children to hospitals were (a) for treatment under general anaesthetic (GA) or relative analgesia (RA) and (b) for restorative care of dentally anxious children. GDPs also referred anxious children requiring simple restorative care and/or RA to the CDS. Only eight GDPs (7%) cited a lack of experience as a reason for referral of dentally anxious children for simple restorative care, compared to 53 (48%) who cited a lack of RA facilities, and 25 (23%) who cited financial considerations. CONCLUSIONS: GDPs refer children to both hospital services and the CDS, and identify a lack of RA facilities and economic pressures as key reasons for referral. PMID- 18397592 TI - Denture cleansers, soft lining materials and water temperature: what is the effect? AB - AIM: To investigate the effect of water temperature, in association with four denture cleansers, on the colour and surface roughness of five soft lining materials. METHOD: Two silicone soft lining materials (Molloplast B and Sofreliner) represented the heat-cured, laboratory-based and chairside materials that are available. The wide range of chairside, acrylic resin-based, temporary soft lining materials was represented by Coe-Soft, Viscogel and Eversoft. An alkaline hypochlorite (Dentural) and three effervescent peroxide solutions with a range of active pH values (Steradent Active Plus, pH 6; Steradent Extra Strength, pH 3; Superdrug Fresh, pH 11) were selected as the cleansers. The denture cleansers were used at two temperatures: 55 degrees C (manufacturers' recommended temperature) and 80 degrees C. The colour of the specimens was evaluated using a colorimeter and the surface roughness was assessed using a three-dimensional, non contact surface profilometer. This study reported on the effect of using both the higher temperatures and the recommended temperature on the colour and roughness of permanent and temporary soft lining materials. RESULTS AND CONCLUSION: The results of the study showed that the temperature of the water used with denture cleansers might have a detrimental effect on the colour and surface roughness of some permanent and temporary soft lining materials. PMID- 18397593 TI - A case of severe caries and demineralisation in a patient wearing an essix-type retainer. AB - This case report presents a patient who developed significant caries and demineralisation due to consumption of large quantities of cariogenic drinks while he was wearing an Essix retainer. The risks to dental health during orthodontic retention and the responsibilities of general dental practitioners, orthodontists and patients are highlighted. PMID- 18397594 TI - Body dysmorphic disorder: a growing problem? AB - Body dysmorphic dsorder (BDD) is characterised by a preoccupation with an imagined defect in one's appearance or, in the case of a minor physical anomaly, the individual's concern is markedly excessive, causing significant distress in their life. One of the most common areas of preoccupation is the dentofacial region, with up to 20% of patients diagnosed with BDD expressing specific concern regarding their dental appearance. With the increased ability to undertake dental aesthetic and reconstructive procedures, in addition to the use of facial aesthetic procedures, it is paramount for all dental clinicians to have an understanding of this condition. BDD patients often request multiple aesthetic procedures, but remain unsatisfied with their treatment results. It is imperative for the dental clinician to diagnose this condition prior to instigating clinical treatment, and to make an appropriate referral. PMID- 18397595 TI - Is there such a thing as a 'healthy occlusion'?: lessons from history. AB - Opinions on occlusion and its link to oral health could not be more diverse. These opinions are used as justification for an equally diverse range of treatments for so-called occlusally related problems, from heavy wear to chronic orofacial pain. Many claim to have evidence to back up their opinions, although much of it appears to be from a very weak source or based on clinical experience. The strongest case appears to be made by those who believe there is no link, by virtue of a lack of evidence to support its existence. Yet for many clinicians, it is a self-evident truth that occlusion must be related in some way to the function and dysfunction of the stomatognathic system. This article uses historical research to highlight where the pro-occlusionists may be going wrong. It also offers a new concept, based on anthropological studies, that indicates occlusion's link to oral health, and explains why most patients appear to have no occlusal problems for most of the time. This theory needs to be tested by researchers to establish its validity. In the meantime, dentists who feel the need to treat occlusal problems will find it supports a very conservative approach to care. PMID- 18397596 TI - An initial analysis of the research competency level of the Faculty of General Dental Practice (UK)'s Divisional Research Contacts. An in-house survey. AB - PURPOSE: The purpose of this survey was to identify the current level of competence of the Faculty of General Dental Practice (UK)'s [FGDP(UK)'s] Divisional Research Contacts (DRCs). METHODOLOGY: The FGDP(UK)'s DRCs were each sent a copy the Faculty's Research Competencies Framework (RCF). A mutually convenient time for a telephone interview was then agreed between the DRCs and one of the Faculty's two National Research Facilitators. During the interview, a structured questionnaire was used and the DRCs' qualifications and research experience were recorded along with their perceived knowledge and skill levels for each of the 21 competencies in the RCF, using a five-point Likert scale. RESULTS: Eighteen of the 20 DRCs were interviewed, of whom 12 either possessed or were studying for higher research or partresearch degrees (PhD or MSc). All had some previous research experience. For 11 of the 15 competencies in the four domains of practical skills, problem-solving, attitudes and dissemination, DRCs rated their knowledge and skills levels as average or higher. For two of these 11 competencies (communication skills and understanding the need for research to support clinical practice), all 18 DRCs rated their levels of knowledge and skills as good or excellent. In the domain of roles and functions, DRCs rated their levels of knowledge and skills at a lower level. Seven rated their competence in designing and implementing a series of studies that address a significant issue and in writing research funding applications to major bodies as poor or very poor. CONCLUSIONS: The present survey has provided the basis for a future, in-depth training needs analysis along with an insight into the research competency level of 18 of the Faculty's 20 DRCs. PMID- 18397599 TI - Ernest Gutmann heritage, 30 years after. PMID- 18397600 TI - Ernest Gutmann (1910-1977). PMID- 18397601 TI - The Denervated Muscle: 45 years later. AB - Forty-five years after its publication, Ernest Gutmann's book, The Denervated Muscle, still stands as a landmark publication. It summarized the state of knowledge of the time and introduced many new researches that were continuing at the Institute of Physiology in Prague. At the time, the response of a muscle to denervation was viewed primarily through the lens of the neurotrophic theory. Advancements in our understanding of neurotrophic effects and mechanisms would now call into question some of the hypotheses and interpretations presented in the book, but many of the research findings have stood the test of time. This review will cover some of the questions asked and data presented in this book, and will place them into the context of contemporary muscle biology. PMID- 18397602 TI - Walking performance, medical outcomes and patient training in FES of innervated muscles for ambulation by thoracic-level complete paraplegics. AB - OBJECTIVE: To discuss functional electric stimulation (FES) gait training of upper motoneuron spinal cord injured complete paraplegics considering ambulation performance, physiologic and metabolic responses as well as psychologic outcome, while providing myologic insight into ambulation via FES when training starts many years post-injury. METHODS: Transcutaneous FES using the Parastep stimulation system, gait training methods with and without major emphasis on muscle reinforcement, cardiovascular and respiratory conditioning. Examination of myofiber tissues and correlation of normal muscles histology versus innervated muscles of upper motor neuron and of denervated muscles of lower motor neuron paraplegics. RESULTS: Published works in literature reviewed in this paper report average walking distance of 440 m/walk when major muscle reinforcement and preconditioning cardiovascular and respiratory systems precedes gait training, versus average 115 m/walk when undergoing direct gait training. Medical, metabolic and psychologic outcomes, as reported in several works, point to benefits of FES walking, including 60% increase in blood flow to lower extremities. Myofiber tissues of patients with upper motor neuron paralysis compare well with those of normal tissue even many years post-injury, while adipose tissue substitute muscle fibers in patients with lower motor neuron lesions. DISCUSSION: Transcutaneous FES allows considerably longer walking distances and speed at the end of training when training involves an extensive pre-conditioning program than with direct gait training. Medical and psychologic benefits are observed, especially concerning blood flow to the lower extremities. Myofiber examinations provide myologic understanding of effectiveness of FES many years post-injury. PMID- 18397603 TI - Local expression of mIgf-1 modulates ubiquitin, caspase and CDK5 expression in skeletal muscle of an ALS mouse model. AB - OBJECTIVE: The functional connection between muscle and nerve is often altered in several neuromuscular diseases, including amyotrophic lateral sclerosis (ALS). Knowledge about the molecular and cellular mechanisms involved in the restorative reactions is important to our understanding of the processes involved in neuromuscular maintenance. We previously reported that muscle-restricted expression of a localized Igf-1 isoform maintained muscle integrity, stabilized neuromuscular junctions, reduced inflammation in the spinal cord and enhanced motor neuronal survival in SOD(G93A) mice, delaying the onset and progression of the disease. In this study, we analysed potential molecular pathways that are modulated by mIgf-1 to counteract muscle wasting and to preserve motor neurons activity. METHODS: We performed molecular and morphologic analysis to address the specific proposed questions. RESULTS AND DISCUSSION: Ubiquitin expression and caspase activity resulted markedly increased in SOD(G93A) muscle but maintained at very low levels in the SOD(G93A) x MLC/mIgf-1 (SOD(G93A)/mIgf-1) transgenic muscle. In addition, CDK5 expression, a serine-threonine protein kinase that has been implicated in a number of physiologic processes in nerve and muscle cells, was reduced in SOD(G93A) muscle but increased in SOD(G93A)/mIgf-1 muscle. Notably, while the toxic p25 protein accumulated in SOD(G93A) muscle, no accumulation was evident in the SOD(G93A)/mIgf-1 muscle. The maintenance of muscle phenotype was also associated with maintenance of a normal peripheral nerve, and a greater number of myelinated axons. CONCLUSION: These observations offer novel insights into the role of mIgf-1 in the attenuation of muscle wasting in the mouse model of ALS disease. PMID- 18397604 TI - Atrophy-resistant fibers in permanent peripheral denervation of human skeletal muscle. AB - OBJECTIVE: Human muscle fibers usually undergo severe atrophy/degeneration as a result of long-term peripheral denervation. However, some biopsies from paraplegic patients suffering complete conus cauda syndrome display the presence of a small percentage of muscle fibers with a very large diameter (big fibers). The objective of the present study is to determine if these big fibers are the result of residual innervation/reinnervation, or if instead they are fibers resistant to atrophy. METHODS: Human muscle biopsies were harvested from the vastus lateralis of spinal cord injury (SCI) patients affected by complete lower motor neuron lesion (LML). The specimens were either processed for light microscopy or embedded for electron microscopy (EM). RESULTS: Our results indicate that the big fibers are neither the results of residual innervation or sparse reinnervation. In spite of the fact that the extrasynaptic NCAM immunostaining disappear a few months after SCI, the big fibers are characterized by positive molecular markers of denervation, that is, the differential labeling of their dystrophin molecule by anti-C and anti-N terminals antibodies. Furthermore, the EM analysis shows that these cells present the peculiar ultrastructural disarrangements of the contractile apparatus and of the internal membrane systems characteristic of 'peripheral denervation'. No fibers presenting large areas of cross-striation were found. The EM analysis provides the final evidence that these big fibers are muscle fibers which are indeed denervated, very different from normal and/or disused (e.g. upper motor neuron lesion) muscle fibers. DISCUSSION: Although these large muscle fibers are surprisingly more frequent in human muscle biopsies after 3 years from SCI than earlier, it remains to be determined whether their presence in some biopsies but not in others is caused by sampling, or is related to other factors such as to subjects' background genetics, or the extent of passive stretching induced by different rehabilitation strategies. PMID- 18397605 TI - Expression of myositis specific autoantigens during post-natal myogenesis. AB - Idiopathic inflammatory myopathies such as polymyositis (PM) and dermatomyositis (DM) are a group of rare autoimmune diseases, characterized by an inflammatory infiltrate within the skeletal muscle and high titer of circulating autoantibodies in the patient's serum. The etiopathogenesis of these diseases is not known and the relationship between the specific muscle involvement and the ubiquitary presence of the targeted antigens is still unclear. The enhanced expression of myositis specific autoantigens in regenerating muscle fibers from biopsies of PM and DM patients compared to normal muscle has been recently demonstrated. In order to understand whether candidate autoantigens in myositis are expressed during post-natal myogenesis, we performed immunolocalization studies of myositis specific autoantigens in skeletal muscle from newborn and adult rats. Our observations indicate the presence of myositis specific autoantigens during post-natal myogenesis, with possible implications for the induction and/or amplification of the immune-inflammatory response, in patients affected with autoimmune myositis. PMID- 18397606 TI - Hypoxia: the third wheel between nerve and muscle. AB - Skeletal muscles not only obey carefully all motor commands received via motor nerves from nervous system, but also are ready to modify their structure and function to be more suited to the tasks assigned by nervous system. Thus, nervous system appears as the major modulator of the muscle structure and function. Other factors, however, may interfere with the nerve-muscle partnership and among them, hypoxia plays a pivotal role because skeletal muscles exhibit a great variability of the oxygen fluxes and because hypoxia per se has a powerful influence on muscle fibers. The adaptation of skeletal muscles to nerve-induced activity is particularly evident with low frequency tonic patterns and examples are given by chronic low frequency stimulation and by endurance training. Adaptation includes fiber type transitions towards a slow-oxidative phenotype, increased mitochondrial density and increased capillary/fiber ratio. Hypoxia can trigger some of such changes and this has suggested that low oxygen tension at fiber level might be a mediator, possibly based on HIF and VEGF, of the muscle adaptation to increased contractile activity. Chronic hypoxia can, however, induce opposite modifications, such as a fiber type transition from slow oxidative to fast-glycolytic and mitochondrial loss. In such conditions, the increased contractile activity can antagonize hypoxia effects. Thus, hypoxia can play a double role in the nerve-muscle relationship, either reinforcing the nerve influence or antagonizing it. This short review aims to re-examine the ambiguous relationships between nerve-induced contractile activity and hypoxic conditions and to suggest possible interpretations of the double role played by hypoxia. PMID- 18397607 TI - Mechano-sensitivity of normal and long term denervated soleus muscle of the rat. AB - INTRODUCTION AND OBJECTIVES: Evidence showed that physical forces, as passive stretching or active contraction, may counteract various kinds of skeletal muscle atrophy due, for instance, to muscle immobilization, pathophysiology or denervation. Accordingly, active muscle contraction induced by functional electric stimulation is helpful to reduce the muscle atrophic state in denervated man. Moreover, there is evidence that also passive mechanical stimulation of the sarcolemnic membrane may reduce the atrophic muscle state. As to the mechanisms by which mechanical stimulation modulates muscle physiology and pathophysiology, there is a growing list of facts that signaling pathway to the nucleus involves stretch activated channels (SACs) of the sarcolemma and the cytoskeleton. SACs activation allowed a Ca(2+) inflow that activates Ca(2+)-dependent molecular signals. Cytoskeleton may be activated by Ca(2+)-dependent and -independent paths and its contraction and elongation represent not only a mechanical signal to the nucleus but also a stimulus for many molecular signals. The aim of this work was to evaluate in soleus muscle of the rat, the mechano-sensitivity of SACs before and after medium and long term denervation. METHODS: Electrophysiologic experiments were made in normal and denervated Soleus muscle of Wistar rats. Currents were recorded in voltage clamp by intracellular microelectrodes inserted in a single fiber. RESULTS: Our findings demonstrated that SACs were expressed in normal soleus muscle and that SAC currents were potentiated by muscle stretching. Another important result was that the sensitivity to stretching increased after denervation and was particularly evident in long term denervated muscles. DISCUSSION: The reported effects are in agreement with the effects of exercise on inducing muscle hypertrophy or with the positive effects on repairing the atrophic state of skeletal muscles by mechanical stimulation or, in denervated humans, by the functional electrical stimulation (FES). PMID- 18397608 TI - Skeletal muscle is enriched in hematopoietic stem cells and not inflammatory cells in cachectic mice. AB - OBJECTIVE: Cachexia, a debilitating syndrome characterized by skeletal muscle wasting, is associated to many chronic diseases and diminishes the quality of life and survival of patients. Tumor-derived factors and proinflammatory cytokines, including TNF-alpha, IL-6 and IL-1 beta, mediate cachexia. In response to elevated cytokine levels, increased proteasome-mediated proteolysis and auto phagocytosis result in muscle wasting. The histologic features of muscle cachexia are not fully elucidated. Therefore, we analysed alterations of different cell populations in cachectic muscle. METHODS: By immunohistochemical and cytological approaches, we characterized changes in the abundance of cellular populations in the musculature of a murine model of cancer cachexia (C26-bearing mice). RESULTS: Cachectic muscle displayed a decreased DNA content proportional to muscle mass wastage. A decrease in the number of nuclei occurred in the muscular but not in the stromal compartment. Cachectic muscle showed: mild modulation of myeloperoxidase activity, a neutrophil marker; reduction of macrophages in the endomysium; decrease in CD3(+) lymphocyte number. Conversely, a statistically significant enrichment in Sca-1(+) CD45(+) hematopoietic stem cells (HSCs) occurred in cachectic muscle. DISCUSSION: The elevated levels of cytokines which characterize cachexia may represent a trigger for inflammatory cell activation. However, we find that in cachexia, inflammatory cells in muscle are not increased while muscle tissue nuclei decline. Our data suggest that the inflammatory cell mediated stress is not an etiologic component of muscle wasting in cachexia. The relative increase in HSCs in cachectic skeletal muscle suggests an attempt to maintain muscle homeostasis by recruitment and/or activation of stem cells. PMID- 18397609 TI - Influence of locomotor training on the structure and myosin heavy chains of the denervated rat soleus muscle. AB - OBJECTIVE: Mechanism of denervation atrophy remains poorly understood. In particular, the question about irreversibility of the late atrophy is still open. Therefore, in the present study, we investigated whether and how a passive movement can affect a progress of atrophy in rat soleus muscle. To address this issue, a locomotor training on a treadmill was applied to rats with their right hindlimb muscles denervated. METHODS: The hindlimb muscles were denervated by cutting the sciatic nerve. Starting either 7 days or 1 month after the surgery, the animals were trained on a treadmill. Two months after denervation, the soleus muscle was investigated using light and electron microscopy and biochemical methods. Control soleus muscles were obtained from non-trained animals: the untreated and the 2-month denervated. RESULTS: Locomotor training caused slight increase in denervated rat soleus muscle weight and significant increase in its fiber diameter. The training positively affected some of the factors that were believed to be the reasons of atrophy irreversibility, because of significant increase in the number of capillary blood vessels and muscle fiber nuclei with the concomitant decrease in the number of severely damaged muscle fibers and amount of collagen. Morphology of the contractile apparatus was also improved as more regular organization of sarcomeres and the hexagonal arrangement of myosin filaments was evident. Moreover, the amount of myosin heavy chains (MHC) significantly increased after training. The effects were more evident in the animals with longer training. CONCLUSION: Passive movement seems to attenuate some of the pathologic processes within the denervated muscle. PMID- 18397610 TI - Embryonic chick cocultures of neuronal and muscle cells. AB - OBJECTIVE: The aim of this report was to set up an effective experimental model of cocultures between cells from spinal cord explants and myotubes from adductor muscle. METHODS: We obtained neuronal cells from chick spinal cord explants at embryonic day 5 (ED5) by means of an enzymatic digestion. Small spinal cord fragments were added in cultured muscle cells committed to the differentiative program. Myoblasts were isolated from the chick adductor muscle at ED12. RESULTS AND DISCUSSION: The validation of the experimental model was confirmed by a remarkable spreading pattern of neuronal cells, labeled with a NF200 antibody, and high concentration of myotubes, marked by alpha-actinin antibody. The indication of neuronal contacts was highlighted by the alpha-bungarotoxin. This communication reports one of the few morphologic description of muscular and neuronal coculture preparation, performed on chick embryos. CONCLUSION: The experimental model presented in this work might be a useful tool to study the cascade of myogenic positive and negative signals activated by paracrine neuronal factors. PMID- 18397611 TI - Embryonic neurons transplanted into the tibial nerve reinnervate muscle and reduce atrophy but NCAM expression persists. AB - OBJECTIVE: The aim of this study was to use the glycogen depletion technique to determine whether reinnervated muscle fibers could be distinguished from denervated muscle fibers by their size or by neural cell adhesion molecule (NCAM) expression. METHODS: Medial gastrocnemius muscles of five adult Fischer rats were reinnervated from embryonic neurons transplanted into the distal stump of the tibial nerve. Ten weeks later, the transplants were stimulated repeatedly to deplete reinnervated muscle fibers of glycogen. Areas of reinnervated (glycogen depleted) muscle fibers were measured and assessed for NCAM expression. The areas of muscle fibers from reinnervated, denervated (n=5) and unoperated control muscles (n=5) were compared. RESULTS: Mean reinnervated muscle fiber area was significantly larger than the mean for denervated fibers (mean +/- SE: 40 +/- 6 and 10 +/- 1% of unoperated control fibers, respectively). NCAM was expressed in 55 +/- 7% of reinnervated fibers (mean +/- SE; range: 42-77%). The mean areas of reinnervated fibers that did or did not express NCAM were similar. NCAM was only expressed in some fibers in completely denervated muscles. DISCUSSION: Our data show that NCAM expression does not differentiate muscle denervation or reinnervation. Quantifying the area of large fibers did distinguish reinnervated muscle fibers from denervated fibers and showed that reinnervation of muscle from neurons placed in peripheral nerve is a strategy to rescue muscle from atrophy. PMID- 18397612 TI - New 3D hyaluronan-based scaffold for in vitro reconstruction of the rat sciatic nerve. AB - OBJECTIVE: For peripheral nerve regeneration, three-dimensional distribution and growth of cells within the porous scaffold are of clinical significance. The purpose of this study was to test in vitro a novel hyaluronic acid-based tubular conduit (HYAFF-11 biomaterials: 1 x 10 mm) as a nerve guide. METHODS: Human fibroblasts, RN22 Schwann cell lines, human umbilical vein endothelial cells and primary nerve cells, obtained from neonatal rat sciatic nerve, were harvested and seeded on HYAFF-11 devices. Histologic (hematoxylin-eosin), immunohistochemical (antibodies to S100, CD31 and Von Willebrand factor) and PCR analyses were performed after 7 and 14 days from cell seeding onto biomaterials. MTT-based (thiazolyl blue) and DELFIA cell proliferation kit tests were performed to observe the biocompatibility of the cells cultured within the biomaterial devices. RESULTS: We concluded that the conduits were not cytotoxic and demonstrated that cultured RN22 Schwann cells and rat Schwann cells grow in vitro on new artificial nerve conduits. We thus inferred that the HYAFF-11 conduit was a suitable biomaterial able to support nerve cell growth in vitro and after 14 days of cultivation, remained circular with a round lumen, maintaining the size and shape of its original architecture. Finally, attachment and proliferation of endothelial cells attested to the feasibility of developing a coculture system to promote in vivo integration of a microvascularized nerve substitute. DISCUSSION: HYAFF-11 pre-seeded with Schwann and endothelial cells has the potential to be an alternative to autografting for the repair of long peripheral nerve defects. PMID- 18397613 TI - Persistence of regenerative myogenesis in spite of down-regulation of activity dependent genes in long-term denervated rat muscle. AB - Contrary to general expectation, in humans, we have recently shown that after complete conus cauda lesion, the lower motoneuron denervated myofibers may survive several years. In adult rats, the sciatectomized muscle progresses in 4-6 months from severe atrophy to a dystrophic stage and undergoes a dramatic weight loss; during this process, myofiber death/regeneration processes maintain a decreasing population of very small, but vital myofibers. At the same time, in vitro electrophysiologic recordings show that denervated fibers can maintain membrane excitability longer than they can retain contractile properties. A certain level of myofiber regeneration seems to have a role in the process, with the early re-expression of embryonic subunits of integrins and acetylcholine receptor subunits. In the present work, using the reliable real-time quantitative PCR, we confirm the long-lasting occurrence of myoblast proliferation-dependent events and their focal nature. In fact, we show here that in sciatectomized muscle, the expression of 12 selected genes was differentially regulated after 3 and 9 month denervation. At both time points, indexes of muscle activity/inactivity and tissue remodeling (proteolysis, energy usage and angiogenic factors) were down-regulated, while indexes of regenerative myogenesis (Myogenin, MyoD, MRF4 and MHCemb) were up-regulated. Immunohistochemistry with anti-MHCemb and anti-NCAM monoclonal antibodies show that such regeneration events were focally distributed. We conclude that myofiber regeneration is a non compensatory mechanism, which prolongs the chance of reinnervation during long lasting denervation. It may also contribute to muscle recovery in paraplegic patients, even when rehabilitation strategies based on functional electric stimulation start late after spinal cord injury (SCI). PMID- 18397614 TI - Electrophysiologic stimulation improves myogenic potential of muscle precursor cells grown in a 3D collagen scaffold. AB - The production of engineered three-dimensional (3D) skeletal muscle grafts holds promise for treatment of several diseases. An important factor in the development of such approach involves the capability of preserving myogenicity and regenerative potential during ex vivo culturing. We have previously shown that electrical stimulation of myogenic cells grown in monolayer could improve the differentiation process. Here we investigated the effect of exogenous electrical field, specifically designed to mimic part of the neuronal activity, on muscle precursor cells (MPCs) cultured within 3D collagen scaffolds. Our data showed that electric stimulation did not affect cell viability and increased by 65.6% the release rate of NO(x), an early molecular activator of satellite cells in vivo. NO(x) release rate was decreased by an inhibitor of NO synthase, both in stimulated and non-stimulated cultures, confirming the endocrine origin of the measured NO(x). Importantly, electrical stimulation also increased the expression of two myogenic markers, MyoD and desmin. We also carried out some preliminary experiments aimed at determining the biocompatibility of our seeded collagen scaffolds, implanting them in the tibialis anterior muscles of syngeneic mice. Ten days after transplantation, we could observe the formation of new myofibers both inside the scaffold and at the scaffold/muscle interface. Altogether, our findings indicate that electrical stimulation could be a new strategy for the effective 3D expansion of muscle precursor cells in vitro without losing myogenic potential and that 3D collagen matrices could be a promising tool for delivering myogenic cells in recipient muscles. PMID- 18397615 TI - [Laparoscopic splenectomy]. PMID- 18397616 TI - [Laparoscopic splenectomy in children with hereditary spherocytosis]. AB - Based on the positive international experience with laparoscopic splenectomy on a large number of patients since 1991, we treated two teenagers with symptomatic hereditary spherocytosis and moderate splenomegaly. Both operations and postoperative courses were uneventful, and the children had improved clinically at the one-month control. The aim of this article is to emphasize the good results with the operation worldwide and suggests that the method may be introduced in Denmark. PMID- 18397617 TI - [Endovascular repair of aortic aneurysm a paradigmatic shift]. AB - The evolution of endovascular aortic repair (EVAR) is rapid and global. The aim is to replace open surgery as a life-saving treatment. According to randomised controlled studies, abdominal EVAR is a better option than open repair in terms of safety and efficacy in the short run. However, the overall survival in EVAR patients with severe morbidity is not influenced since mortality in this group is mainly unrelated to the aneurysm. Endovascular repair is also a solid alternative in thoracic pathologies and aortic ruptures. Patient selection and follow-up in EVAR is mainly governed by vascular imaging, especially 3-D CT reconstructions. There is a need for an increased number of endovascular specialists in Denmark since an increasing number of patients are suitable for EVAR treatment. PMID- 18397618 TI - [Mutations in the heart's pacemaker channels--a new cause of sick sinus node syndrome and long-QT syndrome]. AB - The sinus node hyperpolarization-activated If current generated by the cardiac pacemaker channels HCN2 and HCN4 determines the autonomous beating of the heart. Cardiac arrhythmias, like long-QT syndrome, are often caused by irregularities of the heart action potential generated by mutations in cardiac ion channel genes. Mutations in the HCN4 gene have been associated with sick sinus syndrome and long QT syndrome. The identification of cardiac arrhythmia disease-associated genes makes possible new therapeutic strategies based on gene-specific drug treatment and gene therapy. PMID- 18397619 TI - [Should patients with chronic renal insufficiency be treated with folic acid, vitamin B6 and vitamin B12?]. AB - Hyperhomocysteinemia is associated with CKD and increased cardiovascular risk. Treatment with folic acid and possibly vitamin B12 reduces homocysteine in renal insufficiency. At present there is no evidence of reduced mortality or morbidity following such treatment. This was confirmed in a recent study evaluating combined treatment with folic acid, B6 and B12 in chronic renal insufficiency and ESRD. While an ongoing study in renal transplant patients is anticipated, there is not sufficient evidence currently to recommend treatment of all chronic renal patients with folic acid, B6 or B12. PMID- 18397620 TI - [Temperature measurement]. PMID- 18397621 TI - [Prevalence and prognostic significance of pathological glucose metabolism in acute myocardial infarction]. AB - Individuals with diabetes are at increased risk of developing acute myocardial infarction (AMI) and recent investigations have shown that impaired glucose tolerance (IGT) contributes to a comparable increased risk of AMI. Studies in which oral glucose tolerance test (OGTT) has been used to diagnose IGT/diabetes report a prevalence of unrecognized pathological glucose metabolism in 54-66% of admitted patients. Newly detected diabetes by OGTT, but not IGT, also correlates strongly with prognosis after AMI even after adjusting for traditional risk factors. PMID- 18397622 TI - [Prevalence of glucometabolic abnormalities in connection with myocardial infarction in a Danish coronary care unit]. AB - BACKGROUND: The prevalence of previously undiscovered glucometabolic abnormalities such as diabetes and impaired glucose tolerance (IGT) has been shown to be high among selected groups with myocardial infarction. The aim of this study was to establish the prevalence of known diabetes and undiagnosed glucometabolic abnormalities in an unselected population with myocardial infarction (MI) at admission and after 3 months by use of oral glucose tolerance testing (OGTT). MATERIALS AND METHODS: During an 18-month period patients consecutively admitted with acute MI in a Danish coronary care unit were included. Out of the 218 patients included, 44 had known diabetes. Using the results of OGTT and fasting venous plasma glucose measures, the remaining 174 patients were classified in glucometabolic categories. 80 out of 111 invited patients were classified by OGTT after 3 months. RESULTS: 71% of the cohort had abnormal glucose metabolism, 29% IGT, 22% newly detected diabetes and 20% established diabetes. There were no significant differences in body mass index, blood pressure, cholesterol, HDL, LDL or triglycerides in the groups without known diabetes. Patients with abnormal glucose metabolism were older than their normal glucose tolerance counterparts. There was a trend towards normalization of glucometabolism after 3 months. CONCLUSION: The prevalence of abnormal glucose metabolism in MI-populations is high. Among individuals with presumably normal glucose tolerance 37% were classified as IGT and 27% as diabetics. Association between common risk factors for both abnormal glucose regulation and MI could not explain these findings. OGTT is probably required for correct diagnosis of abnormal glucose regulation in patients with MI. PMID- 18397626 TI - [Surgical treatment of pulmonary embolism]. AB - This case report describes three cases of acute massive pulmonary embolism treated with surgical embolectomy at Skejby Sygehus (Aarhus University Hospital), Denmark, in the period from March 2005 to January 2006. It was necessary to open each pulmonary artery in all three patients in order to remove as much of the embolic material as possible. The three patients survived the operation, although one had hypoxic brain damage resulting from cardiac arrest prior to hospitalization. They all had normal echocardiography postoperatively. Surgical embolectomy is therefore a feasible treatment for acute massive pulmonary embolism. PMID- 18397623 TI - [Thrombo-prophylaxis in Danish surgical departments. Status 2005 and 25 years' development]. AB - OBJECTIVES: To assess Danish surgical departments' attitudes to and clinical guidelines on the use of perioperative thrombo-prophylaxis (TP). MATERIALS AND METHODS: Questionnaires regarding TP were sent to Danish departments which performed major surgery in 2005. The questionnaires contained questions regarding the use of TP routinely after clinical guidelines for both elective and acute surgery, indications for the use of TP, the type of TP and the duration of the TP treatment. The results are registered descriptively. The results are compared with five previously performed surveys from 1981-2000. RESULTS: In the five surveys from 1981 to 2000, 85%, 90%, 92%, 95%, 93% answered the questionnaire, in the present survey 88% answered. Since the beginning of these surveys, there has been a significant increase in the use of TP following clinical guidelines from approximately 50% in 1981 to 96% of the departments performing major surgery in 2005. In acute surgery we found a significant increase in the number of departments using TP following clinical guidelines from 43% in 2000 to 81% in 2005 (p<0.01 Fishers exact test). All departments that used TP employed low molecular weight heparin. The use of prolonged TP was sporadic. CONCLUSION: 30 years after the first evidence in favour of TP was published, 96% of surgical departments which perform major surgery use TP routinely based on international clinical recommendations. A large number of departments use TP in connection with acute surgery in comparison with previous surveys. A future task is more focus on the use of prolonged TP after major surgical procedures. PMID- 18397627 TI - [Multicystic peritoneal mesothelioma is an uncommon lesion that is most often discovered incidentally]. AB - Symptoms are unusual. The lesion is benign with a favourable prognosis but local recurrence is reported not to be rare, and in a few cases malignant transformation has been described. PMID- 18397628 TI - [Somatostatin analogues in the treatment of bleeding GI-angiodysplasias]. AB - Angiodysplasias account for 2-8% of GI-bleeding episodes. We describe a 75-year old patient with a history of anaemia for 1 year before being diagnosed with multiple angiodysplasias in the GI-tract. The patient needed weekly blood transfusions until he started treatment with octreotide. After 4 months of treatment the need for transfusions had decreased dramatically and faeces was negative for blood. Treatment was switched to long-acting octreotide (Sandostatin Lar). The case illustrates the usefulness of somatostatin analogues in the treatment of bleeding GI-angiodysplasias. PMID- 18397629 TI - [Weight loss prolongs life. Danish Association for the Study of Obesity]. PMID- 18397630 TI - [Emergency medicine--a developing specialty. The Danish Society for Emergency Medicine]. PMID- 18397632 TI - [Specialty training courses in anesthesiology. The Danish Society of Anesthesiology and Intensive Medicine]. PMID- 18397631 TI - [Danish General Medicine Database--future tool! The Danish Society of General Medicine]. PMID- 18397633 TI - [Stress--a new challenge. The Danish Society of Occupational Medicine]. PMID- 18397634 TI - [Hereditary Parkinson disease. The Danish Society of Movement Disorders]. PMID- 18397635 TI - [Atypical type 1 diabetes. The Danish Society for Child and Adolescent Diabetes]. PMID- 18397636 TI - [Psychosomatic symptoms in young children. The Danish Society of Child and Adolescent Psychiatry]. PMID- 18397637 TI - [Drug-eluting stents in percutaneous coronary intervention. The Danish Society of Cardiology]. PMID- 18397638 TI - [Nonmelanoma skin cancer. The Danish Society of Dermatology]. PMID- 18397639 TI - [Genetics in non-autoimmune diabetes. The Danish Society of Endocrinology]. PMID- 18397640 TI - [Treatment of medically intractable epilepsy. The Danish Epilepsy Society]. PMID- 18397641 TI - [The campaign for promotion of patient safety and quality of health care. The Danish Society of Public Health and the Danish Society for Patient Safety]. PMID- 18397642 TI - [Short-bowel syndrome treated with glucagon-like peptide. 2. The Danish Society for Gastroenterology]. PMID- 18397643 TI - [Future elderly will be one of the greatest challenges. The Danish Geriatric Society]. PMID- 18397644 TI - [Tuberculosis infection burden in Greenland. Greenland Medical Society]. PMID- 18397645 TI - [Danish clinical hepatoepidemiology. The Danish Society of Hepatology]. PMID- 18397646 TI - [Targeted treatment of mouth cancer. The Danish Society for Head and Neck Surgery]. PMID- 18397647 TI - [A new therapeutic principle for migraine]. PMID- 18397648 TI - [Is there an unrecognized epidemic of primary aldosteronism? The Danish Society of Hypertension]. PMID- 18397649 TI - [Tyrosine kinase inhibitors in the treatment of polycythemia vera and related conditions. Danish Society of Hematology]. PMID- 18397650 TI - [Central mechanisms in peripheral nerve injuries. Danish Society for Surgery of the Hand]. PMID- 18397651 TI - [Q fever--a new zoonotic disease in Denmark? The Danish Society for Infectious Diseases]. PMID- 18397652 TI - [The sepsis campaign--increased survival with sepsis bundles. The Danish Society for Intensive Care]. PMID- 18397653 TI - [Interventional radiology. Danish Society of Interventional Radiology]. PMID- 18397654 TI - [Debate on specialist education--again. The Danish Surgical Society]. PMID- 18397655 TI - [Genomic microarray technology. The Danish Society of Clinical Biochemistry]. PMID- 18397656 TI - [Disease-related malnutrition--an evidence-based phenomenon. The Danish Society of Clinical Nutrition]. PMID- 18397657 TI - [Children on the drug development map. The Danish Society of Clinical Pharmacology]. PMID- 18397658 TI - [Lung scintigraphy--SPECT/CT. The Danish Society of Clinical Physiology and Nuclear Medicine]. PMID- 18397659 TI - [Has the new technology made Gram's staining method redundant? The Danish Society of Clinical Microbiology]. PMID- 18397660 TI - [New diagnostics of neuromuscular diseases. The Danish Society of Clinical Neurophysiology]. PMID- 18397661 TI - [Osteoporosis--new therapeutic possibilities. The Danish Bone Society]. PMID- 18397662 TI - [Quality in focus. The Danish Society for Health Sector Quality]. PMID- 18397663 TI - [Improved therapies in nephrology. The Danish Society of Nephrology]. PMID- 18397664 TI - [Multiple sclerosis--the significance of genes. The Danish Society of Neurology]. PMID- 18397665 TI - [Fewer twin pregnancies after fertilization in vitro. The Danish Society of Gynecology and Obstetrics]. PMID- 18397666 TI - [New treatment of age-related macular degeneration. The Danish Society of Ophthalmology]. PMID- 18397667 TI - [Thyroid surgery--a growing challenge. The Danish Society of Otolaryngology, Head and Neck Surgery]. PMID- 18397668 TI - [Emergency cancer investigation requires quick tissue specimen handling. The Danish Society of Pathologic Anatomy and Cytology]. PMID- 18397669 TI - [Reference programs for anxiety disorders and unipolar depression. The Danish Society of Psychiatry]. PMID- 18397670 TI - [The new CT scanners. The Danish Society of Radiology]. PMID- 18397671 TI - [Center for abused children. The Danish Society of Forensic Medicine]. PMID- 18397672 TI - [Rheumatology in 2008. The Danish Society of Rheumatology]. PMID- 18397673 TI - [Challenges of the antithrombotic treatment. The Danish Society for Thrombosis and Hemostasis]. PMID- 18397674 TI - [Does male circumcision prevent HIV/AIDS? The Danish Society for Tropical Medicine and International Health]. PMID- 18397675 TI - [New therapies for urologic cancer diseases. The Danish Society of Urology]. PMID- 18397676 TI - First-Line management of CML: a state of the art review. AB - With the advent of imatinib, an inhibitor of the fusion Bcl-Abl tyrosine kinase that is responsible for the disease phenotype, first-line treatment of chronic myelogenous leukemia (CML) has undergone rapid changes. As a consequence of the success of imatinib, allogeneic stem cell transplantation (allo-SCT) has moved from the first-line indication to an option that is usually considered for patients without optimal response to imatinib or those for whom treatment with imatinib has failed. First-line use of interferon-alpha (IFN-alpha), with or without cytarabine or hydroxyurea, is now rarely considered. Most patients with newly diagnosed chronic-phase CML experience complete cytogenetic responses during imatinib treatment, but continued treatment is necessary to prevent cytogenetic or molecular relapse. Cumulative evidence with imatinib and IFN-alpha show that experiencing early and deep responses is important in preventing progression to more advanced phases of the disease. These features emphasize the need to carefully monitor patients for residual disease and provide guidance on treatment options in the event of imatinib intolerance or failure. This article reviews the current role of allo-SCT, imatinib, and IFN-alpha in treating newly diagnosed chronic-phase CML, highlighting the benefits and challenges of long term patient management, and discusses emerging trends. PMID- 18397677 TI - Resistance and relapse with imatinib in CML: causes and consequences. AB - The 60-month update of the Immediate Risk-Stratification Improves Survival (IRIS) study, a trial of standard-dose imatinib in patients with newly diagnosed chronic phase chronic myelogenous leukemia, showed an 89% projected rate of overall survival. Although this is an enormous improvement over all previously available therapies, approximately 16% of patients experienced failed therapy with imatinib and 7% progressed to accelerated phase or blast crisis. Therefore, resistance continues to be a clinical problem. Exploiting the full potential of imatinib requires close attention to patient response to recognize suboptimal response and treatment failure as early as possible and to make timely adjustments to the therapeutic strategy. This article reviews the definitions and mechanisms of primary and acquired imatinib resistance. Particular emphasis is placed on point mutations in the kinase domain of BCR-ABL as a mechanism of drug resistance, because they highlight important issues related to drug design and the biology of CML. Given the wealth of publications on the subject, including all information was not feasible and choices had to be made. PMID- 18397678 TI - Beyond dose escalation: clinical options for relapse or resistance in chronic myelogenous leukemia. AB - The development of imatinib has changed the management of chronic myeloid leukemia (CML), producing high response rates in most patients. However, most individuals treated with imatinib, 400 mg, have residual molecular disease, and both intrinsic and acquired resistance can occur. The newer tyrosine kinase inhibitors, dasatinib and nilotinib, are effective in patients with imatinib resistant CML, In patients with relapse or resistance, current guidelines recommend escalating the dose of imatinib or switching to new tyrosine kinase inhibitors. Dasatinib has been investigated in patients who were resistant or intolerant to imatinib. Switching to dasatinib, 70 mg, twice daily has been shown to be more effective than high-dose imatinib. Another study found that dasatinib, 100 mg, once daily was just as effective as the twice-daily regimen but was better tolerated. Nilotinib is also effective in most patients with resistance or intolerance to imatinib and is associated with minimal toxicity. Other inhibitors, such as bosutinib and INNO-406, are being developed with favorable early results. New drugs are still needed, particularly for individuals who are resistant to tyrosine kinase inhibitors or those with the T3151 mutation. Emerging CML therapies, some of which have different mechanisms of action from those of tyrosine kinase inhibitors, have shown promising results and could offer an alternative to these patients as monotherapy or in combination. PMID- 18397679 TI - Advanced CML: therapeutic options for patients in accelerated and blast phases. AB - Tyrosine kinase inhibitor (TKI) therapy has impacted the natural course of chronic myelogenous leukemia (CML), because patients diagnosed as having chronic phase disease can experience long-lasting responses. However, for patients with advanced CML (accelerated and blast phases), the efficacy of all current therapies is reduced. For these patients, allogeneic stem cell transplantation remains the preferred treatment if a donor is available, although TKIs play a valuable role as a bridging therapy. For patients with accelerated-phase CML, imatinib, dasatinib, and nilotinib have been shown to produce meaningful rates of hematologic and cytogenetic response. Imatinib and dasatinib are also approved for blast-phase CML. Studies with the newer agents have involved heavily pretreated patients; however, response rates have been at least comparable to those achieved with imatinib in previous studies. Therefore, these newer, more potent TKIs will probably be more likely to induce a deep response in previously untreated patients. Moreover, because fewer mechanisms appear to exist for secondary resistance to dasatinib and nilotinib, reducing the potential for disease to escape TKI therapy, these agents may result in improved longer-term outcomes. However, BCR-ABL-independent pathways may also become more important, indicating that other therapeutic targets may also have a future role in managing patients with advanced CML. PMID- 18397680 TI - BCR-ABL tyrosine kinase inhibitors in chronic myeloid leukemia: using guidelines to make rational treatment choices. AB - The success of the BCR-ABL tyrosine kinase inhibitor (TKI) imatinib in improving prognosis in chronic myeloid leukemia (CML) has led to its wide use as first-line therapy at a standard dose of 400 mg daily. As more patients have undergone therapy, the development of molecular and clinical resistance to imatinib has raised further therapeutic challenges. The 2 main approaches to overcoming resistance are imatinib dose escalation and the use of alternative more potent TKIs, such as dasatinib or nilotinib. The phase II SRC/ABL Tyrosine Kinase Inhibition Activity Research Trials (START) of dasatinib have established dasatinib as potent and effective in overcoming imatinib resistance or intolerance in all phases of CML. The most recent treatment guidelines by the National Comprehensive Cancer Network now contain recommendations for using dasatinib in this setting. The issue of when to change from imatinib to an alternative agent in preference to imatinib dose escalation is keenly debated, particularly as new clinical evidence emerges, which highlights the importance of achieving early cytogenetic and molecular responses for a good long-term outcome. Identifying patients in whom a change to dasatinib or nilotinib is more appropriate than imatinib dose escalation is therefore important. PMID- 18397681 TI - Healthy subjects volunteering for Phase I studies: influence of curiosity, exploratory tendencies and perceived self-efficacy. AB - OBJECTIVE: To test the hypothesis that trait-curiosity and perceived self efficacy influence the willingness of healthy subjects to volunteer for participation in Phase I studies. MATERIALS AND METHODS: A group of healthy subjects who had never participated in clinical studies ("index group") were invited to participate in a Phase I study. They were assessed with regard to trait curiosity (Curiosity and Exploration Inventory; CEI-T) and perceived self efficacy (Self-Efficacy Scale; SES) and subjects who accepted the invitation to participate were compared with those who refused and with a group of healthy subjects who had previously participated in clinical studies ("validation group"). RESULTS: A significant positive correlation was found between the willingness to participate and the CEI-T total score (R=0.28; p<0.01), exploratory tendencies (R=0.34; p<0.001), SES total score (R=0.30, p<0.01), initiative and persistence (R=0.29, p<0.01), planning/goal setting (R=0.19, p<0.05) and social self-efficacy (R=0.29; p<0.01). The "index group" subjects who accepted the invitation to participate showed significantly greater CEI-T exploratory tendencies (Z=-3.334, p = 0.001, Mann-Whitney test) and total scores (Z=-2.703, p<0.01) and greater SES total score (Z=-3.131, p<0.01), initiative and persistence (Z=-3.065, p<0.01), planning/goal setting (Z=-2.173, p<0.05) and social self-efficacy (Z=-2.954, p<0.01) than subjects who refused. No differences were found between the subjects in the "index group" who accepted the invitation and subjects in the "validation group". Using a logistic regression model, both CEI-T exploratory tendencies and SES initiative/persistence were significant predictors of participation. CONCLUSION: Subjects higher in curiosity/exploration and in perceived initiative/persistence are more willing to volunteer for Phase I studies. The impact of these self-selection biases on Phase I study results is unknown but deserves further evaluation. PMID- 18397682 TI - Effect of renal impairment on the pharmacokinetics of eslicarbazepine acetate. AB - OBJECTIVE: Antiepileptic drugs are often used in patients with some degree of renal impairment. The objective of this study was to evaluate the effect of renal function on the pharmacokinetics of eslicarbazepine acetate (ESL, formerly known as BIA 2-093), a new antiepileptic drug under clinical development. METHODS: ESL pharmacokinetics following 800 mg single dose was characterized in subjects with normal renal function (n=8, control group), and in patients with mild renal impairment (n=8), moderate renal impairment (n=8), severe renal impairment (n=8), and end-stage renal disease requiring hemodialysis (n=8). RESULTS: ESL suffered extensive first-pass hydrolysis to eslicarbazepine (S-licarbazepine), the main active metabolite. While eslicarbazepine Cmax did not significantly differ between the different groups, the extent of systemic exposure, assessed by AUC, increased when renal function decreased. Eslicarbazepine CL/F and CLR were, respectively, 3.40 l/h and 1.04 l/h (17.3 ml/min) in the control group, and 2.10 l/h (35.0 ml/min) and 0.61 l/h (10.2 ml/min) in the mild, 1.60 l/h (26.7 ml/min) and 0.22 l/h (3.7 ml/min) in the moderate, and 1.33 l/h (21.2 ml/min) and 0.09 l/h (1.5 ml/min) in the severe renal impairment groups. Although the total amount of eslicarbazepine recovered in urine until 72 h post-dose was similar in the control and mild renal impairment groups, a decrease was found in the moderate and severe renal impairment groups. Major metabolites recovered in urine were eslicarbazepine and its glucuronide form. Clearance of minor metabolites (R licarbazepine, oxcarbazepine and their glucuronides) was also dependent on renal function. In patients with end-stage renal disease, dialysis was effective in removing the ESL metabolites from the circulation. CONCLUSIONS: ESL metabolites are excreted primarily by renal route and their clearance is dependent on renal function. ESL dosage adjustment may be necessary in patients with a creatinine clearance <60 ml/min. PMID- 18397683 TI - Calcium balance during calcitriol and paricalcitol administration in healthy humans. AB - OBJECTIVE: Secondary hyperparathyroidism in hemodialysis patients requires optimal correction of vitamin D deficiency with active vitamin D and analogues. It has been postulated that new vitamin D analogues, i.e. paricalcitol, efficiently suppress parathyroid hormone serum levels (PTH), but do not increase intestinal calcium absorption as much as calcitriol. The effects of calcitriol and paricalcitol on calcium balance can best be characterized under standardized conditions in healthy individuals with normal renal function, because the urinary calcium excretion at steady state corresponds to the net calcium absorption in the gut. METHODS: In a randomized, double-blind, placebo-controlled, 3-way crossover Phase I study in 13 healthy individuals we investigated the changes compared to placebo in PTH and urinary calcium excretion during 6-day treatment periods with paricalcitol (1.5 microg/day) and calcitriol (0.5 microg/day). RESULTS: 24-hour urinary calcium excretion was stable during 6 days of placebo administration. Neither paricalcitol nor calcitriol significantly changed calcium excretion. Urinary creatinine, magnesium and phosphate excretion also remained unchanged over the study periods irrespective of the treatment. However, calcitriol was shown to be effective in reducing iPTH levels during 6 days of treatment (mean reduction 4.03+/-0.69 pmol/l), whereas paricalcitol had no effect. CONCLUSION: Using a dosing ratio of 1:3 for calcitriol:paricalcitol, i.e. the same conversion factor used previously in studies on hemodialysis patients, only calcitriol was able to reduce iPTH levels in healthy individuals. Low-dose calcitriol reduced iPTH levels without raising calcium absorption and without including any hypercalcemia. PMID- 18397684 TI - Hemodynamic effects of dobutamine in patients with an exacerbation of chronic systolic heart failure treated with low doses of carvedilol. AB - Although most patients in Europe with systolic heart failure (SHF) are treated with I(2)-blocking agents at doses significantly lower than the recommended dose, there is limited information available regarding the hemodynamic effects of dobutamine in this patient population. Therefore, a study was carried out in patients (n=31) admitted to the University Hospital, Larissa, Greece with an acute exacerbation of chronic SHF (25 men and 6 women, mean age 58 years, range 32 a 80 years, left ventricular (LV) ejection fraction 95%) and continuous suppression of platelet COX-1 may represent an exception. Apart from aspirin, which irreversibly inactivates COX-1, high-dose naproxen causes sustained COX-1 inhibition throughout the dose interval in some individuals. The present study examines whether naproxen sodium after a single dose administration and at steady state using "over-the-counter (OTC) doses" produces sufficient COX-1 inhibition. COX-2 inhibition was assessed concomitantly. METHODS: Ex vivo inhibition of COX enzymes and the pharmacokinetics of naproxen were assessed in four volunteers receiving 220 mg naproxen sodium b.i.d. for 7 days. Blood samples were obtained pre-dose, at specified time points after the first dose on Day 1, and 12 hours after the previous evening dose on Days 2, 3, 4, 5 and 8. Recovery was assessed up to 36 hours after the last dose. Coagulation-induced thromboxane B2 formation and lipopolysaccharide-induced prostaglandin E2 synthesis were measured ex vivo in human whole blood as indices of COX-1 and COX-2 activity. RESULTS: Maximal inhibition after a single dose and at steady state were as follows: 94% and 93% (COX-1), and 79% and 85% (COX-2). A greater than 95% COX-1 inhibition was observed transiently in 2 of 4 volunteers at the time of maximal plasma concentration after a single-dose administration and in 1 of 4 volunteers throughout the 12-hour dose interval at steady state. For both isoenzymes, COX inhibition correlated with naproxen plasma levels (ex vivo IC50 values of 35.48 micromol/l (COX-1) and 64.62 micromol/l (COX-2)). CONCLUSIONS: Administration of naproxen sodium at OTC doses was associated with a profound inhibition of both COX enzymes. Although low-dose naproxen may elicit a virtually complete COX-1 inhibition in some individuals, it does not mimic the reliable, sustained and complete COX-1 blockade produced by aspirin. In conclusion, prolonged treatment with 220 mg naproxen sodium b.i.d. is not expected to provide sufficient cardioprotection in all patients, but may influence platelet function in some. PMID- 18397692 TI - Acarbose alleviates rise in plasma uric acid concentration induced by sucrose ingestion. AB - Sucrose is divided by alpha-glucosidase into fructose and glucose, which are considered to raise plasma uric acid concentration through purine degradation and/or decreased uric acid excretion. AIMS: We investigated the effect of acarbose, an alpha-glucosidase inhibitor, on the increased plasma concentration of uric acid caused by sucrose. METHODS: 6 healthy males were studied. After an overnight fast, sucrose at 1.5 g/kg was ingested. Urine was collected 1 hour before sucrose ingestion and then twice at 1-hour intervals after ingestion. Blood was taken twice, at the midpoint of each 1-hour period. 2 weeks later, the same protocol was followed, with acarbose at 100 mg added at the beginning of the sucrose ingestion. RESULTS: Sucrose ingestion raised the plasma concentration of uric acid by 10%, whereas with the addition of acarbose the rise in plasma concentration of uric acid was reduced (p < 0.01) without changes in urinary uric acid excretion and fractional uric acid clearance. Urinary excretion and fractional clearance of oxypurines were unchanged in both experiments. CONCLUSIONS: Acarbose is considered to alleviate the rise in plasma concentration of uric acid induced by sucrose by inhibiting its absorption since no changes in uric acid excretion and fractional clearance were observed. PMID- 18397693 TI - Septic shock and community-acquired pneumonia associated with etanercept therapy. AB - OBJECTIVE: To report a case of septic shock and community-acquired pneumonia in a patient with psoriatic arthritis receiving treatment with etanercept. PATIENT DETAILS: A 65-year-old woman diagnosed as having psoriatic arthritis had received treatment with etanercept. Chest X-ray studies were normal and the tuberculin skin test was negative. Two months after etanercept therapy, the patient presented to our emergency department with fever, cough, chest pain and generalized weakness. Chest radiography revealed a right pulmonary infiltrate. Her condition rapidly deteriorated and she went into shock with a further drop in her blood pressure, tachycardia and tachypnea. She was intubated, mechanically ventilated and was treated with fluids, cardioversion and amiodarone. Empiric therapy with levofloxacin, amikacin and cefepime were initiated. In the urinalysis, the result of a rapid test for Streptococcus pneumoniae was positive. Etanercept treatment was suspended due to a possible adverse reaction associated with this drug. At the start of therapy her clinical condition improved slowly. On Day 28, the patient was afebrile and she was discharged from the intensive care unit. DISCUSSION: Most of the infections associated with etanercept therapy have been reported in patients with rheumatoid arthritis. Based on our observations, etanercept was the possible offender in the development of septic shock and respiratory failure in community-acquired pneumonia. There was a temporal relationship between exposure to the drug and onset of symptoms. Etanercept was the only drug administered before the septic shock developed. Based on the Naranjo algorithm, the adverse reaction could be considered possible. CONCLUSION: Patients initiated on etanercept should be counseled and receive appropriate screening before drug initiation. All febrile and newly occurring concomitant illnesses should be promptly evaluated. General practitioners should discontinue etanercept treatment and institute prompt and aggressive intervention if infection develops. PMID- 18397694 TI - Use of fondaparinux as an anticoagulant during hemodialysis: a preliminary study. AB - OBJECTIVE: To study the effect of fondaparinux, a new antithrombotic agent, as an anticoagulant during a 4-hour conventional hemodialysis session. materials and methods: Fondaparinux was administered as an anticoagulant to 16 chronic hemodialysis patients during a single 4-hour hemodialysis session at an intravenous bolus dose of 2.5 mg. Eight patients were using high-flux polyester polymer alloy (PEPA) dialyzers (Group A) and the remainder low-flux polysulfone dialyzers (Group B), whilst all had received conventional doses of tinzaparin sodium as an anticoagulant during the previous month. The dialyzers were primed with 1 l of normal saline containing 5,000 IU of unfractionated heparin. Blood samples for the measurement of INR, APTT (activated partial thromboplastin time) and anti-Xa levels were taken before the study dialysis session (pre), 5 min postdialysis (post), and before the next dialysis session (next). Mean fibrin/clot formation in the extracorporeal circuit and dialyzer was assessed macroscopically by visual inspection and was graded using a 4-point scale. RESULTS: Predialysis anti-Xa levels were 0.04 A+/- 0.03 IU/ml in Group A, and 0.025 A+/- 0.025 IU/ml in Group B (p = NS). Postdialysis anti-Xa levels were significantly higher than predialysis levels in both groups (Group A = 0.16 A+/- 0.04 IU/ml, Group B = 0.46 A+/- 0.12 IU/ml, p < 0.02 for both) and significantly higher in Group B compared to Group A (p < 0.025). Anti-Xa levels before the next dialysis session were 0.06 A+/- 0.04 IU/ml in Group A and 0.25 A+/- 0.06 IU/ml in Group B (p < 0.0001 between Groups A and B). APTT values were significantly higher in postdialysis than predialysis samples for both groups (higher by 27.0 A+/- 26.0% in Group A and 24.3 A+/- 31.9% in Group B). No significant differences were found when comparing APTT values in pre, post and next samples between Groups A and B. No differences were also found between pre, post and next samples for INR values, either within or between groups. Mean fibrin/ clot formation score in the extracorporeal circuit at the end of the study dialysis session was significantly higher in patients of Group A than those of Group B (p < 0.05). Dialysis had to be terminated before the completion of 4 hours in 2 patients of Group A because of the presence of extensive fibrin/clots in the circuit and dialyzer. CONCLUSIONS: Our findings indicate that fondaparinux sodium at an intravenous dose of 2.5 mg can be used successfully as an anticoagulant during a 4-hour conventional hemodialysis session in patients dialyzed with low-flux polysulfone dialyzers, but not in those dialyzed with high-flux dialyzers. However, anti-Xa levels in the former patients were still increased before the next dialysis session, potentially exposing the patients to an increased risk of bleeding. PMID- 18397695 TI - A bioequivalence study comparing two formulations of lopinavir/ritonavir capsules. AB - This investigation was carried out to evaluate the bioavailability of a new single fixed-dose combination formulation of lopinavir and ritonavir, relative to reference product, Kaletra (133.3 mg lopinavir/33.3 mg ritonavir) capsules, manufactured by Abbott Laboratories, Chicago, IL, USA. The bioavailability study was carried out on 72 healthy male and female volunteers who received a single dose of 3 capsules (133.3 mg lopinavir/33.3 mg ritonavir) of the test (T) and the reference (R) products in the fasting state, in a randomized, balanced, 2-way crossover design. After dosing, serial blood samples were collected for a period of 72 hours. Plasma harvested from blood was analyzed for lopinavir and ritonavir by a sensitive and validated simultaneous liquid-chromatographic and mass spectrometric (LC-MS/MS) assay. Mean oral clearance (Cl/F) values of the FDC were 4.92 and 23.54 l/h for lopinavir and ritonavir, respectively, the maximum plasma concentrations (C(max)), area under the plasma concentration-time curve up to the last measurable concentration (AUC(0-t)), and to infinity (AUC(0-infinity)), were analyzed statistically under the assumption of a multiplicative model. The time to maximum concentration (t(max)) was analyzed assuming an additive model. The parametric confidence intervals (90%) were calculated by Schuirmann's two 1-sided t-test criteria. It was found that the test/reference (T/R) ratios for the pharmacokinetic parameters AUC(0-t), AUC(0-infinity) and C(max) (after initial log transformation) were well within the bioequivalence acceptance range of 80 125% as per international regulatory guidelines. Therefore, the two formulations were considered to be bioequivalent [Food and Drug Administration 2003]. PMID- 18397696 TI - Hypervolemia, arterial hypertension and cardiovascular disease: a largely neglected problem in peritoneal dialysis. AB - Here we review the existing data on hypertension, volume overload and volume control in peritoneal dialysis (PD) patients and comment on the impact of these factors on residual renal function and cardiovascular disease in PD patients. PMID- 18397697 TI - Prevalence of cryoglobulinemia and autoimmunity markers in renal-transplant patients. AB - AIMS: To examine the prevalence of cryoglobulinemia (Cryo) and autoimmune markers in renal-transplant recipients in a stable condition, and to determine its risk factors and impact upon allograft function. PATIENTS AND METHODS: In May, 2006, 117 kidney-transplant (KT) recipients, aged 31 a 76 years, were tested for cryoglobulinemia, hepatitis B and C, complement C3, C4, CH50, antinuclear (ANAs), anticytoplasmic nuclear (ANCAs) and anticardiolipid antibodies, rheumatoid factor (RF), and lymphocyte subpopulations. Renal, liver, and hematological tests were also performed. Immunosuppressive regimens were based on calcineurin inhibitors (82%). RESULTS: Cryo was positive in 47 patients (Cryo(+): 40.2%), of whom 13 were HCV+ (27.7%), with characteristics of Type II in 21.2% and Type III in 78.8%. Cryo was positive in 13/16 (81.2%) of HCV+/RNA+ patients vs. 34/101 (33.6%, p = 0.0003) of HCV-negative patients. Cryo(+) RT patients had been recipients of a graft for longer (142 months) than Cryo(-) patients, i.e., 95 months (p = 0.02). Creatinine clearances were similar in the two groups (56 vs. 50 ml/mn, p = 0.5), as were microalbuminuria and albuminemia. There was no difference between Cryo(-) and Cryo(+) patients in terms of age, sex, HLA mismatch, daily steroid doses, liver and hematological tests, ANAs, anticardiolipid antibodies, serum complement, and lymphocyte subpopulations. RF occurred in all Cryo(+) patients and in 82.8% of Cryo(-) patients, with higher titers in the Cryo(+) group (23 vs. 9 UI/ml, p = 0.012). ANCA occurred in nine Cryo(-) but in no Cryo(+) patients (p = 0.013). Finally, a multivariate analysis was not able to determine any predictive factor associated with cryoglobulinemia. CONCLUSION: Cryoglobulinemia is frequent after KT, and is associated with HCV markers, RF, and absence of ANCA. PMID- 18397698 TI - A single-center experience with BK virus nephropathy. AB - BACKGROUND: BK virus nephropathy has an increasing role in renal transplant dysfunction, since new, highly potent immunosuppressive drugs have been introduced into therapy following renal transplantation. Diagnosis of acute impairment of renal transplant function is complicated by difficulty in differentiating BK virus nephropathy from acute rejection. PATIENTS AND METHODS: We retrospectively described the findings and therapeutic approaches of 6 consecutive patients with BK virus nephropathy in our transplantation center (75 80 transplantations/ year). BK virus nephropathy was classified according to Drachenberg et al. [2004]. RESULTS: We observed an incidence rate of < 1% for BK nephropathy in our center. Four patients had a pattern B whereas 2 patients revealed a pattern C of BK virus nephropathy. Focal C4d-positive staining of peritubular capillaries were found in 2 of the 6 cases. For earlier detection of BK nephropathy, a diagnostic algorithm for each patient after renal transplantation was established. Urine was continuously monitored by cytology for decoy cells and PCR for BK virus DNA. If PCR was also positive for the BK virus in plasma, biopsy of the renal allograft was performed. Thereby diagnosis could be confirmed sooner. For treatment of BK nephropathy in our center, we reduced immunosuppressive agents and initiated a virustatic treatment with cidofovir in the first 3 cases. However, results were not satisfactory and two allografts were lost. We then reconsidered our therapeutic approach and switched the immunosuppressive treatment to leflunomide with consistent low dose steroids. We use therapeutic drug monitoring for leflunomide and aim at a target level of 40 - 100 microg/ml. We lost no allograft with BK nephropathy since using this therapeutic approach. CONCLUSION: In our center, leflunomide therapy, but not cidofovir, was effective in patients with BK virus nephropathy of the renal allograft. PMID- 18397699 TI - Postdialysis outcomes associated with consistent anemia treatment in predialysis patients with chronic kidney disease. AB - AIMS: Anemia and cardiovascular (CV) events are major complications of chronic kidney disease (CKD) during dialysis. We conducted a retrospective observational study in CKD patients with anemia to evaluate the association between predialysis use of erythropoiesis-stimulating agents (ESAs) and postdialysis CV outcomes. METHODS: The study analyzed claims data on incident hemodialysis patients aged > or = 18 years (identified between January 2000 and November 2005). Patients were identified as anemic and ESA-treated prior to dialysis. ESA treatment was categorized into 4 consistency groups (from least to most consistent ESA use). RESULTS: Of 5,848 hemodialysis patients, 52% were identified as anemic prior to onset of dialysis. Predialysis ESA treatment was received by 62% of anemic patients, with only 23% receiving the most consistent treatment. The risk of a CV event was significantly lower for the ESA-treated compared with ESA-untreated patients (relative risk (RR) 0.70, 95% (95% confidence intervals (CI) 0.61 - 0.82)). Compared with ESA-untreated, those who received ESAs had significantly lower risk of acute myocardial infarction (RR 0.65 (95% CI 0.44 - 0.95)) or inpatient mortality (RR 0.52 (95% CI 0.40 - 0.68)). ESA-treated patients in each of the 4 consistency groups had significantly lower risk of CV events compared with ESA-untreated patients, with the greatest benefit seen in patients who received most consistent ESA (RR 0.61 (95% CI 0.48 - 0.76)). CONCLUSIONS: This analysis suggests consistent ESA use to treat anemia of CKD in the predialysis period is associated with improved cardiovascular outcomes in postdialysis patients. PMID- 18397700 TI - Idiopathic retroperitoneal fibrosis: a role for mycophenolate mofetil. AB - PURPOSE: Idiopathic retroperitoneal fibrosis (IRPF) is an unusual progressive illness for which consistent therapeutic recommendations have not been devised. The present report describes a collaborative nephrology and urology approach to distinguish IRPF from secondary disease and then combine necessary acute surgical or radiological intervention with short-term corticosteroid and with mycophenolate mofetil (MM) to facilitate steroid tapering and long-term management. MATERIALS AND METHODS: 21 patients have been evaluated and followed over a 7-year period, 16 with characteristic IRPF and 5 with secondary retroperitoneal disease. IRPF patients initially received high-dose corticosteroid and MM. We report clinical follow-up along with imaging studies of the retroperitoneum and related organs, serologic markers for systemic disease, and nonspecific acute-phase reactants as indicators of ongoing disease activity. RESULTS: Among IRPF patients, uniform success in stabilizing clinical signs and symptoms, radiological disease in the retroperitoneum and associated organs, and inflammatory indicators have been observed. Corticosteroid therapy can be limited to 6 months or less and MM to approximately 2 years, all with substantial impact on the natural history of IRPF. CONCLUSIONS: This is not a randomized, controlled trial, and patients were often referred with prior complications and/or treatments, however, the systematic approach and consistent results support the utility of MM as a safe and effective choice for long-term stabilization in IRPF. PMID- 18397701 TI - An assessment of cinacalcet HCl effects on bone histology in dialysis patients with secondary hyperparathyroidism. AB - AIMS: Cinacalcet lowers plasma parathyroid hormone (PTH) levels in patients with secondary hyperparathyroidism (sHPT), but the bone histologic response has not been described. This prospective, double-blind, placebo-controlled trial assessed the effects of cinacalcet on bone histology and serum markers of bone metabolism in dialysis patients with sHPT. METHODS: Patients with intact PTH (iPTH) > or = 300 pg/ml were randomly assigned 2:1 to receive cinacalcet or placebo with concurrent vitamin D and/or phosphate binder therapy. Cinacalcet (30 - 180 mg/day) was used to achieve iPTH levels < or = 200 pg/ml. Bone biopsies were performed before and after one year of treatment. RESULTS: Baseline and end-of study data were available from 32 patients (19 cinacalcet, 13 placebo). Baseline bone turnover was elevated in 27, reduced in 3 and normal in 2 patients. Serum bone-specific alkaline phosphatase (BSAP) and N-telopeptide (NTx) were elevated. Cinacalcet treatment decreased PTH and diminished activation frequency, bone formation rate/bone surface, and fibrosis surface/bone surface. Adynamic bone was observed in three patients receiving cinacalcet; in two of these, PTH levels were persistently low (< 100 pg/ml). The histomorphometric parameter changes in bone corresponded to PTH, BSAP and NTx reductions. Bone mineralization parameters remained normal. CONCLUSIONS: Treatment with cinacalcet lowered PTH and reduced bone turnover and tissue fibrosis among most dialysis patients with biochemical evidence of sHPT. PMID- 18397702 TI - In-center daily on-line hemodiafiltration: a 4-year experience in children. AB - Our daily dialysis program was started in September 2002: in-center daily on-line hemodiafiltration (DIH) was carried out in 3-hour sessions, 5 - 6 times weekly, on-line assessment KT/Vurea of minimal 1.5 per session, polysulfone membranes. 12 children were included: median age 7.4 years (2.10 - 16.8 years), renal residual function less than 3 ml/min/1.73 m2 (Kcreat + Kurea/2), vascular access central catheter (n = 4) or fistula (n = 8), 7/12 being converted from peritoneal dialysis to DIH. Median follow-up on DIH was 11 months (4 - 43 months), endpoint was kidney transplantation (11/12) or transfer to another center (1/12). Monthly assessments of dialysis parameters (KT/Vurea, predialysis phosphatemia), diet survey (3 consecutive days), medications (number of antihypertensive drugs, phosphate chelators, potassium chelators) and statural growth were performed. At start of DIH, diet intake due to medical prescription and limited appetite was restrictive with limitation in water, salt (20 mmol/day), potassium and proteins (median 35 g/day, range 20 - 80 g); only 2/12 children were free of antihypertensive drugs, all received phosphate and potassium chelators, and growth retardation occurred (7/12 in prepubertal children, median height SDS 1.52) despite rhGH therapy (5/12 patients). At the end of DIH, diet was free, protein intake high (2 - 3 g/kg/day, range 30 - 100), 10/12 children were free of antihypertensive drugs, 4/12 received potassium chelators, 1/12 received phosphate chelators. All the prepubertal children at inclusion (n = 7) showed catch-up growth with a median growth rate of 0.8 cm/month (0.5 - 1.6 cm/ month). DIH allowed to maintain predialysis phosphatemia in a low normal range (median 1.23 mmol/l, range 1.65 - 0.63), without (11/12 children) need of phosphate chelators. Thanks to DIH children, parents and team care discovered during DIH a new way of life with motivated children, showing natural compliance (no diet restriction, no or few drugs), and most of all children developing with catch-up of growth. PMID- 18397703 TI - Remission of a B cell CLL-associated membranoproliferative glomerulonephritis Type I with rituximab and bendamustine. AB - In a 56-year-old white male patient, a membranoproliferative glomerulonephritis Type I was diagnosed after a 12-month history of low grade B cell lymphoma (Binet A). HIV, Hepatitis B and C serology were negative. Due to an impairment of renal function despite chemotherapy with COP, an immunochemotherapy consisting of rituximab (6 cycles) and bendamustine (4 cycles) was given. This therapeutic approach caused a complete remission of the nephrotic syndrome. Renal function and arterial hypertension improved markedly. In addition, urinary sediment became normal and proteinuria disappeared completely. PMID- 18397704 TI - Pauci-immune necrotizing and crescentic glomerulonephritis in a patient with systemic lupus erythematosus. AB - We report a case of pauci-immune proliferative necrotizing and crescentic glomerulonephritis in a patient with systemic lupus erythematosus (SLE) who presented with a nephrotic syndrome, while SLE was clinically and serologically quiescent. Immunofluorescence and electron microscopy examination of the kidney biopsy failed to reveal any significant deposit of immunoglobulins as well as of complement C3 and C1q, excluding lupus nephritis as the determinant of crescentic glomerulonephritis. Anti-myeloperoxydase (MPO) as well as anti-proteinase 3 (PR3) antibodies were absent in the serum. An immunosuppressive regimen including corticosteroids and IV cyclophosphamide led to a dramatic decrease of proteinuria. We conclude that necrotizing glomerulonephritis unrelated to lupus nephritis may occur in a patient with quiescent SLE. An underlying dysfunction of cell-mediated immunity might explain the association of pauci-immune crescentic glomerulonephritis and SLE. PMID- 18397705 TI - Successful treatment of nephrotic syndrome due to systemic AL amyloidosis after autologous stem cell transplantation: renal response is an important therapeutic end point. AB - Primary systemic (AL) amyloidosis involves vital organs from the early phase of illness, resulting in poor prognosis. Today, high-dose melphalan followed by autologous peripheral blood stem cell transplantation is an effective treatment for systemic AL amyloidosis. We report a patient with nephrotic syndrome due to systemic AL amyloidosis, who was successfully treated with autologous peripheral blood stem cell transplantation. At follow-up 36 months from ASCT, the patient showed a significant improvement in the signs of peripheral neuropathy and reduction in proteinuria without further organ involvement. Due to poor prognosis with conventional therapy, autologous stem cell transplantation should be considered for treatment in patients with systemic AL amyloidosis, and favorable outcome is ensured with achievement of renal response after ASCT. PMID- 18397706 TI - Rapidly deteriorating renal function with membranoproliferative glomerulonephritis Type 1 associated with hepatitis C treated successfully with steroids and antiviral therapy: a case report and review of literature. AB - INTRODUCTION: The hepatitis C virus (HCV) infection is associated with several renal diseases including mixed essential cryoglobulinemia, membranoproliferative glomerulonephritis (MPGN) and less frequently membranous nephropathy and crescentic glomerulonephritis. We present a case of HCV-associated cryoglobulin negative, MPGN Type 1 with features of early crescents and rapidly deteriorating renal function requiring urgent treatment. CASE: A 35-year-old male was admitted with history of arthralgia and erythematous rash. His past medical history included being an intravenous drug abuser. Biochemistry test showed raised serum creatinine of 150 micromol/l. He had nephrotic range proteinuria of 6 g/day and a serum albumin of 23 g/l. Viral serology for hepatitis B and HIV was negative but confirmed evidence of HCV infection with genotype 3A and viral load of 151,014 copies. He had a renal biopsy and histology demonstrated features of crescentic MPGN Type 1. His renal function deteriorated rapidly with his serum creatinine rising to 300 micromol/l over 2 days. We commenced treatment with intravenous methylprednisolone, 500 mg once daily (o.d.) for 3 days, followed by oral prednisolone 40 mg o.d. Concurrently, pegylated Interferon- (IFN) I+/- was commenced. After a 2-week treatment, his renal function showed remarkable recovery with creatinine reduced to 140 micromol/l. After 3 months, ribavirin was added when his renal function remained stable. He had tolerated his treatment without any major side effects. At 6 months follow-up clinic, his renal function was normal with serum creatinine of 69 micromol/l, 24-h urinary protein had dropped to 0.35 g/day, serum albumin increased to 38 g/l and HCV PCR was negative. DISCUSSION: The current treatment strategy of HCV-associated renal diseases includes targeting viral trigger HCV with interferon and ribavirin. Both IFN-I+/- and ribavirin have their limitation and adverse effects. In a clinical scenario where there is evidence of rapidly deteriorating renal function with crescentic glomerulonephritis, cautious use of immunosuppressive therapy may well be essential in the acute stage to halt the progression of kidney damage. Literature review of the treatment strategy for MPGN Type 1, cryoglobulin negative with early features of crescents associated with HCV showed that there was no report or guideline available. CONCLUSIONS: To our knowledge, this is the first case in the literature of rapidly progressing MPGN Type 1 associated with HCV and nephrotic syndrome treated successfully with antiviral drugs and steroids concurrently. Our case highlights an important treatment strategy and may be beneficial to nephrologists facing this clinical scenario in the future. However, a randomized controlled trial is required to evaluate the efficacy of this treatment combination before it can be a standard treatment. PMID- 18397707 TI - Severe episodes of extra cellular dehydration: an atypical adult presentation of cystic fibrosis. AB - Cystic fibrosis (CF) is usually diagnosed during childhood by respiratory or gastro-intestinal symptoms. Hyponatremic hypochloremic dehydration with metabolic alkalosis is a rare but typical presentation of CF in infants. In contrast, only 3 cases have been described in adults. We report a case of CF in a 33-year-old Caucasian female presenting with a severe sodium and chloride depletion caused by inappropriate sweating. She experienced three episodes of severe dehydration before the diagnosis was suspected. Sweat chloride test was pathological and mild pulmonary involvement was found on CT scan. Delta F508 mutation and a rare mutation (3849+40 A/G) on the intron 19 of CFTR gene were found. Interestingly, our patient has a heterozygote twin sister, carrier of the same mutations of CFTR gene who also developed CF but with a different phenotype. We suspect modifier genes to be implicated in the differences observed between the two phenotypes. We discuss the physiopathology of electrolyte disturbance and review the other similar adults cases. PMID- 18397708 TI - Hypokalemic quadriparesis associated with renal tubular acidosis in a patient with Sjogren's syndrome. AB - Sjogren's syndrome is an autoimmune exocrinopathy that involves both glandular and extra-glandular systems. We report a 25-year-old woman who had rapidly progressive quadriparesis. Biochemical investigations showed severe hypokalemia with hyperchloremic metabolic acidosis diagnosed as distal renal tubular acidosis. Salivary gland biopsy revealed Sjogren's syndrome as the underlying cause. She recovered following from quadriparesis potassium and alkali replacement. PMID- 18397709 TI - The reversed ratio of 1-84 PTH (whole PTH)/intact PTH in a patient on hemodialysis associated with primary hyperparathyroidism. AB - Intact PTH measures not only 1-84 PTH, but also other fragments such as 7-84 PTH. Lately, a measurement of 1-84 PTH has been available as whole PTH assay and the ratio of whole PTH/intact PTH is considered to be between 0.5 and 0.7 in patients on hemodialysis. Therefore, intact PTH should be higher than whole PTH. We present a 57-year-old male with chronic renal failure on hemodialysis whose whole PTH was higher than intact PTH (the reversed ratio of whole PTH/intact PTH). He showed one enlarged parathyroid gland by an ultrasonic test, CT examination and RI subtraction study. After this gland was removed by surgery, the ratio of whole PTH/intact PTH normalized. The size of the resected gland was 22 x 15 x 11 mm. The histologic examination revealed adenoma. This indicates that, if patients with chronic renal failure showed the reversed ratio of whole PTH/intact PTH, the possibility that they could have primary hyperparathyroidism in addition to secondary hyperparathyroidism should be considered. PMID- 18397710 TI - Two consecutive episodes of acute hepatitis C with different genotypes in a hemodialysis patient responsive to PEG-interferon monotherapy. AB - We report the case of a 63-year-old male patient on long-term hemodialysis who suffered two consecutive episodes of persistent hepatitis C virus infection with different genotypes and was successfully treated with pegylated IFN-alpha monotherapy each time. PMID- 18397711 TI - Cardiac arrest following injection of concentrated trisodium citrate. PMID- 18397712 TI - The third World Kidney Day: looking back and thinking forward. PMID- 18397713 TI - Hepatitis C virus-related kidney disease: an overview. AB - Hepatitis C virus (HCV)-infection leads to chronic liver disease, but also to extra-hepatic manifestations, including kidney disease. We provide an overview of HCV-related kidney diseases in non-transplanted and in kidney transplant patients, and their therapies. Membranoproliferative glomerulonephritis, associated with Type 2 cryoglobulinemia, is the predominant Type of HCV-related glomerulonephritis. Membranous glomerulonephritis and focal segmental glomerular sclerosis are less commonly described. HCV infection seems to be linked to Type 2 diabetes mellitus, and might alter the progression of diabetic-related nephropathy. Patients infected by HCV should be annually screened for markers of kidney disease and, similarly, patients with membranoproliferative or membranous glomerulonephritis should be screened for HCV infection. After transplantation, cryoglobulinemia is frequent and is associated with HCV markers. HCV-related kidney disease requires specific treatment. In non-kidney-transplant patients, treatment relies on either only anti-HCV therapy in cases of moderate renal disease, or combined anti-viral and immunosuppressive therapies in cases of severe renal disease, i.e., nephrotic syndrome and/or progressive renal failure, and in diseases that are refractory to anti-HCV therapy. In kidney transplant patients, ribavirin monotherapy could be used cautiously, whereas rituximab might be a treatment of choice in the presence of cryoglobulinemia. In liver-transplant patients, in addition to anti-HCV therapy, rituximab might be also used. PMID- 18397714 TI - Clinical manifestation of gadodiamide-related nephrogenic systemic fibrosis. AB - AIMS: To further characterize the clinical signs and symptoms of nephrogenic systemic fibrosis, a new and serious disease affecting renal failure patients and caused by some Gd-containing contrast agents, including gadodiamide. MATERIAL: 22 cases of gadodiamide-related nephrogenic systemic fibrosis followed at the nephrology department of Copenhagen University Hospital Herlev. METHOD: Retrospective cohort study based on medical records, personal interviews and physical examinations. RESULTS: Typical first signs of the disease were skin discoloration, induration and warmth, itching, constant pain and other neuropathic symptoms localized to the lower legs. First sign appeared in a median of 14 days (range 0 a 53 days) after gadodiamide exposure. Associated early symptoms included sleeplessness and transient, diffuse hair loss. The predominant late symptom was symmetrical skin stiffness of extremities with or without restricted joint motion. Ten of 22 patients (45, 95% CI: 27 a 66%) were severely disabled due to contractures on the average of 29 months after being exposed to gadodiamide. Four patients died (18, 95% CI: 6 a 41). Patients perceived that intensive physiotherapy was effective in limiting disabling contractures. CONCLUSIONS: Signs and symptoms of nephrogenic systemic fibrosis vary over time and between patients. The disease leads to severe disability in a significant proportion of affected patients. Intensive physiotherapy may limit the development of contractures. PMID- 18397715 TI - Renal protection in diabetes: is it affected by glucose control or inhibition of the renin-angiotensin pathway? AB - BACKGROUND: Recent reports indicate increased risk of renal failure with long term use of angiotensin-converting enzyme inhibitors (ACEI) in diabetes. End stage renal disease (ESRD) in diabetes has increased despite ACEI and angiotensin receptor blocker (ARB) use. This study questions renal protection by ACEI or ARB. Our hypothesis is that uncontrolled hyperglycemia is central to diabetic ESRD while tight glucose control is renoprotective. Cultured endothelial cells show morphological damage that increases with duration of exposure to high glucose and is prevented by insulin and more so by a combination of insulin and heparin. METHODS: Findings from individual patients are compared to clinical trial results wherein ACEI or ARB was emphasized as the prime therapy to prevent progression of diabetic nephropathy to ESRD. Serum creatinine (Scr) changes were the main indicator of renoprotective effects in clinical trials. Creatinine clearance (Ccl), an important marker of glomerular filtration rate, was seldom reported. RESULTS: Our observations show that ACEI-treated patients develop progressive renal failure, whereas renal function remains stable with optimum glucose control. Clinical trials showed that reduction of proteinuria, with ACEI, reduces the risk of ESRD. Our studies show that reduction of proteinuria with use of ACEI or ARB parallels a reduction in Ccl, suggesting that a change in proteinuria is related to Ccl changes. Scr changes are small, giving a deceptive view of renal protection. CONCLUSIONS: Our observations find no evidence of renal protection with ACEI or ARB use in diabetes. Laboratory studies and clinical observations suggest that adequate glucose control is the key to renal protection in diabetes. PMID- 18397716 TI - Effect of oral mizoribine pulse therapy for frequently relapsing steroid dependent nephrotic syndrome. AB - AIM: To evaluate the efficacy of oral mizoribine (MZB) pulse therapy given twice a week for frequently relapsing steroid-dependent nephrotic syndrome (FR-SDNS). SUBJECTS: 16 patients with FR-SDNS with a median age of 11.6 years (range 5.1 a 17.8 years) were enrolled in the study. This study was a Phase II trial. METHODS: The dose of MZB was adjusted to achieve a peak blood level of about 3 microg/ml (10.0-19.7 mg/kg/d, maximum total dose 750 mg) in two divided doses given 2 days a week before a meal. The therapeutic benefits of MZB pulse therapy were assessed based on a comparison of the incidence of relapse (times/year) and the required daily dosage of prednisolone (PSL) before and after therapy. RESULTS: The incidence of relapse after therapy was significantly lower than that before therapy (2.4 A+/- 1.6 vs. 3.4 A+/- 1.1 times/year, p < 0.05), and the required daily dosage of PSL after therapy was lower than that before therapy (0.39 A+/- 0.26 vs. 0.47 A+/- 0.24 mg/kg/d; not significant). During the follow-up period, discontinuation of PSL was possible in 6 of 12 patients who showed a decreased rate of relapse after therapy. The age at entry into the study and the peak blood concentration of MZB of these patients were significantly higher than in four patients who did not show a decreased rate of relapse (12.3 A+/- 4.3 vs. 7.9 A+/- 2.6 years, p < 0.05; 3.00 A+/- 0.93 vs. 1.97 A+/- 0.36 microg/ml, p < 0.005, respectively). No adverse effects were observed in any patients. CONCLUSION: Our results show that MZB pulse therapy is effective in decreasing the frequency of relapse and reducing the required PSL dosage in older pediatric patients with FR SDNS. PMID- 18397717 TI - Switch of ESA therapy from darbepoetin-alpha to epoetin-beta in hemodialysis patients: a single-center experience. AB - AIMS: No study has previously investigated a switch from darbepoetin-alpha to epoetin-beta in unselected dialysis patients. Our study determined the intravenous epoetin-beta dose necessary to maintain or to achieve hemoglobin targets after switching from darbepoetin-alpha. METHODS: In our dialysis center, all eligible dialysis patients (n = 90) were switched from darbepoetin-alpha i.v. to epoetin-beta i.v. in 2005. The epoetin-beta dose was calculated according to the recommended European equimolar conversion factor (1 : 200 microg darbepoetin alpha corresponds to 200 IU epoetin-beta. The intraindividual evaluation compared 12 weeks before with 16 weeks after the switch. The dose of the erythropoiesis stimulating agents (ESA) and the hemoglobin levels were analyzed for the whole period and for the last 4 weeks of both treatment periods. RESULTS: During treatment with darbepoetin-alpha, 71% of a total of 90 patients achieved a hemoglobin level > or = 11.0 g/dl. After switching to epoetin-beta, the mean hemoglobin level decreased significantly from 11.4 A+/- 1.0 g/dl to 11.1 A+/- 0.9 g/dl (p = 0.0016) and the percentage of patients with hemoglobin levels > or = 11.0 g/dl fell to 50% (p = 0.00138). Furthermore, the mean required ESA dose increased by 13% from 4,335 A+/- 3,217 IU/week darbepoetin-alpha to 4,885 A+/- 3,077 IU/week epoetin-beta (p = 0.0001). Comparing the last 4 weeks, the ESA dose increased by 17% from 4,583 A+/- 3,391 IU/week darbepoetin-alpha to 5,372 A+/- 3,672 IU/week epoetin-beta (p = 0.0003). CONCLUSIONS: After switching from darbepoetin-alpha i.v. to epoetin-beta i.v., the equimolar epoetin-beta dose was not sufficient to maintain hemoglobin levels with the same efficacy above 11.0 g/dl. Significantly less patients achieved hemoglobin target values as suggested by the EBPG guidelines. PMID- 18397718 TI - Patient and technique survival of diabetics on peritoneal dialysis: one-center's experience and review of the literature. AB - BACKGROUND: Diabetes is the leading cause of end-stage renal disease (ESRD). This retrospective study investigated the long-term patient and technique survival and sought to identify the predictors of mortality in diabetic patients receiving PD. METHODS: Patients, aged 17 years or more who commenced home PD between January 31, 1994, and December 31, 2001 were included. Clinical data were available for 358 patients out of 418 total patients who started PD during this period. They were followed until cessation of PD, death, or to January 31, 2003. Survival probabilities were generated according to the Kaplan-Meier method, and multivariate Cox proportional hazards models were used to assess predictors of survival. RESULTS: A total of 358 patients were enrolled in the study. Among them, 139 patients (38.8%) were diabetics. The 1-, 2-, 3- and 5-year patient survival rates were 91%, 76%, 66% and 47% in diabetics and 94%, 89%, 84% and 69% in non-diabetics, respectively. Median actuarial patient survival for diabetic patients (51.8 months; 95% CI 36.0 a 67.5 months) was significantly shorter than that of non-diabetic patients (log rank 14.117, p < 0.001). Death-censored technique survival rates at 1-, 2-, 3- and 5-year were 90%, 83%, 67% and 58% in diabetic, and 94%, 87%, 77% and 70% in non-diabetic patients, respectively. Similar to patient survival, the median technique survival time was significantly shorter for diabetic patients (63.9 months; 95% CI 35.7 - 92.2 months) than that of non-diabetic patients (log rank 4.884, p = 0.027). Multivariate Cox regression analysis showed that advancing age was the only independent predictor of death in the diabetic patients, whereas higher age and wider pulse pressure were associated with mortality in non-diabetic patients. CONCLUSION: Long-term patient and technique survival for diabetic patients on PD seem to be improved compared to our previous report and other studies. The mortality of diabetic patients was predicted predominantly by advancing age. PD remains a viable form of long-term renal replacement therapy for diabetic patients with ESRD. PMID- 18397719 TI - Patient perceived barriers to treatment of depression and anxiety in hemodialysis patients. AB - Untreated psychiatric illness correlates with increased mortality, reduced quality of life and increased risk of suicide in renal failure patients, but little is known about why these patients fail to seek mental health care. The purpose of this study was to identify the perceived barriers to mental health services in the hemodialysis patient population. The Beck Depression Inventory (BDI) and Beck Anxiety Inventory (BAI) were used to identify the prevalence and severity of depression and anxiety in a group of 179 hemodialysis patients. Of the 103 patients who completed the surveys, 73.8% were African-American and 62.1% were male. Of the 54.4% of patients identified with depression by scoring 10 or greater on the BDI, 34.0% had mild-to-moderate, 12.6% had moderate-to-severe, and 7.8% had severe depression. Only 13.6% of respondents met criteria for anxiety. Each patient was asked to choose from a list of possible barriers, and 71.4% of patients meeting criteria for depression or anxiety identified a barrier to mental health treatment. Of these, over 70% of patients were unaware of symptoms of depression/ anxiety or did not perceive the need for help. Our results indicate a high prevalence of untreated depression in hemodialysis patients. Patient perceptions of the need for therapy present the most significant barriers to identification and treatment. PMID- 18397720 TI - Acute renal failure after antibiotic-impregnated bone cement treatment of an infected total knee arthroplasty. AB - Antibiotic-impregnated cement is used frequently in revision procedures of infected total hip and knee arthroplasties. Local antibiotic treatment is as effective as the use of systemic antibiotics. The purpose of such treatment is to provide high tissue concentrations of antibiotics and minimize systemic toxicity, especially nephrotoxicity. Though antibiotic-impregnated cement is considered safe in terms of nephrotoxicity, two cases that have implicated aminoglycoside impregnated cement in acute renal failure (ARF) after surgery for an infected total knee arthroplasty (TKA) have been reported [Curtis et al. 2005, Van Raaij et al. 2002]. Two more cases of postoperative ARF after use of combined tobramycin- plus vancomycin-impregnated cement, this time in total hip arthroplasty, have been recently reported [Patrick et al. 2006]. We report a case of ARF in a 61-year-old patient with a history of diabetes mellitus and hypertension after treatment of a febrile infection of a TKA with combined gentamicin- plus vancomycin-impregnated cement. The ARF could not sufficiently be attributed to other causes and though serum concentrations of antibiotics obtained from the 8th postoperative day and thereafter were far below the trough levels associated with nephrotoxicity, gentamicin and vancomycin seem to have contributed significantly to ARF in our case. PMID- 18397721 TI - Therapeutic management of a new case of LCAT deficiency with a multifactorial long-term approach based on high doses of angiotensin II receptor blockers (ARBs). AB - Familial lecithin cholesterol acyltransferase (LCAT) deficiency (FLD) is characterized by the appearance of corneal opacity, anemia, proteinuria progressing to chronic renal failure and abnormalities in the composition of plasma lipoproteins. No established therapy currently exists for this condition. We report here a new case of FLD caused by two novel mutations in the LCAT gene in which, for the first time, aggressive therapy with angiotensin II receptor blockers and lipid-lowering drugs showed benefit in blood pressure, lipid abnormalities, proteinuria and also kidney function, probably delaying progression to renal failure. PMID- 18397722 TI - Headache during hemodialysis - an uncommon cause for a common problem. AB - A hemodialysis patient is described who was suffering from headache during his dialysis sessions. This was due to a neovascular glaucoma causing an increase in intraocular pressure (IOP) during dialysis sessions. After several months his headache decreased but his IOP measurements remained high with almost similar pre and post dialysis values. Headache during hemodialysis may be due to glaucoma, but this can disappear with time, along with a disappearance in the increase in IOP during dialysis sessions. PMID- 18397723 TI - Spontaneous bladder rupture in a chronic hemodialysis patient. AB - Spontaneous bladder rupture is very rare. A 67-year-old woman who was nearly anuric and had been on chronic hemodialysis therapy for diabetic end-stage renal disease for 6 years complained of severe low abdominal pain and fever for 2 days. Abdominal computerized tomography and retrograde cystography revealed the extraperitoneal leakage of contrast medium, confirming bladder perforation. Partial cystectomy around the perforation site and repair of the bladder rupture were performed. Microscopic examination of the excised bladder tissue revealed that the bladder mucosa was ulcerated. Severe suppurative inflammation was observed throughout the bladder wall. Antibiotic treatment was continued for 3 weeks postoperatively, and repeated retrograde cystography showed no evidence of contrast extravasation. She was discharged, with no other complications. PMID- 18397724 TI - Clear cell renal carcinoma presenting as a bleeding cyst in pregnancy: inaugural manifestation of a von Hippel-Lindau disease. AB - Von Hippel-Lindau (VHL) disease is a dominant autosomal disorder inducing the development of many tumors, such as hemangioblastomas in the central nervous system and retina, cysts or tumors (benign or malignant) in the kidneys and/or the pancreas. We report the case of a pregnant woman who presented with a voluminous hemorrhagic cyst of the right kidney with an exophytic lesion detected in the lower median part of the cyst wall. As an anamnestic inquiry resulted in a familial history of VHL disease, a screening imaging was performed and detected three medullary hemangioblastomas. Considering the active bleeding of the renal cyst and its potential malignancy, a unilateral nephrectomy was carried out after pregnancy interruption. Histological analysis confirmed a multilocular clear cell renal carcinoma. This case underlines the importance of screening procedures such as abdominal ultrasonography and medullary magnetic resonance imaging in all pregnant women with a familial history of VHL disease. PMID- 18397725 TI - Hyperphosphoremia in kidney failure- salivary phosphate as a marker and possible target. PMID- 18397726 TI - T wave alternans testing before and after hemodialysis. PMID- 18397727 TI - Cinacalcet improves bone mineral density in a renal transplant recipient with persistent hyperparathyroidism. PMID- 18397728 TI - Nest building by birds. PMID- 18397729 TI - ADF/cofilin. PMID- 18397730 TI - Plant tropisms. PMID- 18397731 TI - Theodor Boveri and the natural experiment. PMID- 18397732 TI - DNA methylation is widespread across social Hymenoptera. PMID- 18397733 TI - Imitation recognition in great apes. PMID- 18397734 TI - Use-induced motor plasticity affects the processing of abstract and concrete language. PMID- 18397736 TI - Extended phenotype: nematodes turn ants into bird-dispersed fruits. AB - A recent study has discovered a novel extended phenotype of a nematode which alters its ant host to resemble ripe fruit. The infected ants are in turn eaten by frugivorous birds that disperse the nematode's eggs. PMID- 18397735 TI - Mitosis: moesin and the importance of being round. AB - Ezrin/radixin/moesin proteins link the actin cytoskeleton to the plasma membrane. Two new reports have found that moesin phosphorylation is essential for mitotic cell rounding and identify a new role for cell rounding in spindle assembly. PMID- 18397737 TI - Multimodal integration: visual cues help odor-seeking fruit flies. AB - Olfactory stimuli are uniquely devoid of directional information, so that multimodal cues are typically required for their localization. A clever new experimental paradigm with flying Drosophila has shown that accurate heading into an odor plume requires panoramic visual cues. PMID- 18397739 TI - Social evolution: ant eggs lacking totipotency. AB - Caste plasticity is a hallmark of social insects. However, maternal effects that limit the caste of eggs may be more widespread than previously appreciated, and can have some odd consequences. PMID- 18397738 TI - mRNA export: RNP remodeling by DEAD-box proteins. AB - The DEAD-box protein Dbp5 was thought to remodel ribonucleoproteins and displace proteins from these complexes in an ATP-dependent fashion to allow for mRNA export from the nucleus. A recent study on Dbp5 shows, however, that its ADP bound form may also perform an important function in displacing export factors from mRNA. PMID- 18397740 TI - Innate immunity: squelching anti-viral signalling with NLRX1. AB - Innate immunity to viruses involves receptors such as RIG-I, which senses viral RNA and triggers a signalling pathway involving the outer mitochondrial membrane protein MAVS. Recent work has identified that NLRX1, a member of another class of innate immune receptors, sequesters MAVS away from RIG-I and thereby prevents mitochondrial anti-viral immunity. PMID- 18397741 TI - Cochlear mechanics: no shout but a twist in the absence of prestin. AB - Mammalian hearing is boosted by mechanically active auditory receptor cells, the outer hair cells which amplify the actions of incoming sounds. Recent evidence indicates that the molecular motor that drives this amplification, prestin, may do more than boogie. PMID- 18397742 TI - Chemical coevolution: host-parasite arms race runs hot and cold. AB - Coevolution is a major process operating across biological communities at a range of spatial scales. Rapid ecological change makes it vital that we understand how coevolution proceeds if we are to conserve genetic diversity, combat disease and predict the effects of species invasions. PMID- 18397743 TI - Chromosome segregation: organizing overlap at the midzone. AB - Sets of overlapping microtubules support the segregation of chromosomes by linking the poles of mitotic spindles. Recent work examines the effect of putting these linkages under pressure by the activation of dicentric chromosomes and sheds new light on the structural role of several well-known spindle midzone proteins. PMID- 18397744 TI - Notch in the niche. AB - Notch signaling is essential for hematopoietic stem cell (HSC) formation during embryogenesis, and hitherto it was also thought to be required for HSC maintenance. However, in this issue of Cell Stem Cell, Maillard et al. (2008) demonstrate rather conclusively that inactivation of the Notch pathway in HSCs does not interfere with their self-renewal. PMID- 18397745 TI - Illuminating the black box of reprogramming. AB - Yamanaka and colleagues, in a Science article currently published online, have generated induced pluripotent stem (iPS) cells from liver and stomach cells, suggesting that transcription factor-induced reprogramming is not restricted to particular cell types (Aoi et al., 2008). These results also provide important insight into the mechanistic basis of reprogramming. PMID- 18397746 TI - Cancer: inappropriate expression of stem cell programs? AB - Cancer stem cells (CSCs) are a subpopulation of cancer cells that possess characteristics, including self-renewal, associated with normal stem cells. In this issue of Cell Stem Cell, Wong et al. (2008) define a core embryonic stem cell (ESC)-like gene expression program that may be important for CSC function in multiple epithelial cancers. PMID- 18397747 TI - Building better beta cells. AB - Type 1 diabetes represents an attractive target for human embryonic stem cell (hESC)-based cell replacement therapies. Recently, in Nature Biotechnology, Kroon et al. (2008) reported encouraging progress toward this goal with the development of functional islet-like structures in mice transplanted with hESC-derived pancreatic endoderm. PMID- 18397748 TI - Hematopoietic stem cells in transit--where's the niche? AB - In this issue of Cell Stem Cell, Min et al. (2008) identify Egr1 as a novel hematopoietic stem cell (HSC) transcription factor that regulates proliferation and mobilization of these cells. An improved understanding of Egr1 targets may lead to better strategies to expand populations of HSCs ex vivo for therapeutic applications. PMID- 18397749 TI - International banking: checks, deposits, and withdrawals. AB - In the 10 years that the technology to produce human embryonic stem cell lines has been available, hundreds of lines have been derived in numerous global locations. These cell lines are being used by researchers across diverse scientific fields to investigate the basic biology, clinical potential, and pharmaceutical applications of these cells and their progeny. In this fast-moving and rapidly growing field, how can we ensure that data generated by different laboratories using the same cell lines are comparable, reproducible, and consistent? One suggestion would be to ensure the quality of the "seed stock" material received and used by researchers. Because a number of laboratories worldwide provide stem cell lines to the scientific community, it seems logical to explore the harmonization of practices between distributors to establish cohesive standards and aid the global movement of stem cell lines to the research community. In the future, when these cells arrive in the clinic for therapeutic use, this consensus of "best practice" should ensure the consistency and facilitate the dissemination of these valuable materials. PMID- 18397750 TI - Centralized banks for human embryonic stem cells: a worthwhile challenge. AB - Centralized banking of human embryonic stem (hES) cells is an endeavor that can benefit individual research efforts and enhance international collaboration but is complicated by the fact that the science is rapidly evolving in an environment of heterogeneous laws, guidelines, and ethical standards. Written from the vantage point of regulatory professionals, this article provides an overview of the benefits of and challenges facing hESC banking enterprises in general with a focus on a global centralized banking effort. PMID- 18397751 TI - Mesenchymal stem cells: revisiting history, concepts, and assays. AB - The concept of mesenchymal stem cells has gained wide popularity. Despite the rapid growth of the field, uncertainties remain with respect to the defining characteristics of these cells, including their potency and self-renewal. These uncertainties are reflected in a growing tendency to question the very use of the term. This commentary revisits the experimental origin of the concept of the population(s) referred to as mesenchymal stem cells and the experimental framework required to assess their stemness and function. PMID- 18397752 TI - Lives of a heart cell: tracing the origins of cardiac progenitors. AB - Heart cells are the unitary elements that define cardiac function and disease. The recent identification of distinct families of cardiovascular progenitor cells begins to build a foundation for our understanding of the developmental logic of human cardiovascular disease, and also points to new approaches to arrest and/or reverse its progression, a major goal of regenerative medicine. In this review, we highlight recent clarifications, revisions, and advances in our understanding of the many lives of a heart cell, with a primary focus on the emerging links between cardiogenesis and heart stem cell biology. PMID- 18397753 TI - Module map of stem cell genes guides creation of epithelial cancer stem cells. AB - Self-renewal is a hallmark of stem cells and cancer, but existence of a shared stemness program remains controversial. Here, we construct a gene module map to systematically relate transcriptional programs in embryonic stem cells (ESCs), adult tissue stem cells, and human cancers. This map reveals two predominant gene modules that distinguish ESCs and adult tissue stem cells. The ESC-like transcriptional program is activated in diverse human epithelial cancers and strongly predicts metastasis and death. c-Myc, but not other oncogenes, is sufficient to reactivate the ESC-like program in normal and cancer cells. In primary human keratinocytes transformed by Ras and I kappa B alpha, c-Myc increases the fraction of tumor-initiating cells by 150-fold, enabling tumor formation and serial propagation with as few as 500 cells. c-Myc-enhanced tumor initiation is cell-autonomous and independent of genomic instability. Thus, activation of an ESC-like transcriptional program in differentiated adult cells may induce pathologic self-renewal characteristic of cancer stem cells. PMID- 18397754 TI - Trophoblast differentiation defect in human embryonic stem cells lacking PIG-A and GPI-anchored cell-surface proteins. AB - Pluripotent human embryonic stem (hES) cells can differentiate into various cell types derived from the three embryonic germ layers and extraembryonic tissues such as trophoblasts. The mechanisms governing lineage choices of hES cells are largely unknown. Here, we report that we established two independent hES cell clones lacking a group of cell surface molecules, glycosyl-phosphatidyl-inositol anchored proteins (GPI-APs). The GPI-AP deficiency in these two hES clones is due to the deficiency in the gene expression of PIG-A (phosphatidyl-inositol-glycan class A), which is required for the first step of GPI synthesis. GPI-AP-deficient hES cells were capable of forming embryoid bodies and initiating cell differentiation into the three embryonic germ layers. However, GPI-AP-deficient hES cells failed to form trophoblasts after differentiation induction by embryoid body formation or by adding exogenous BMP4. The defect in trophoblast formation was due to the lack of GPI-anchored BMP coreceptors, resulting in the impairment of full BMP4 signaling activation in the GPI-AP-deficient hES cells. These data reveal that GPI-AP-enhanced full activation of BMP signaling is required for human trophoblast formation. PMID- 18397755 TI - Canonical notch signaling is dispensable for the maintenance of adult hematopoietic stem cells. AB - Gain-of-function experiments have demonstrated the potential of Notch signals to expand primitive hematopoietic progenitors, but whether Notch physiologically regulates hematopoietic stem cell (HSC) homeostasis in vivo is unclear. To answer this question, we evaluated the effect of global deficiencies of canonical Notch signaling in rigorous HSC assays. Hematopoietic progenitors expressing dominant negative Mastermind-like1 (DNMAML), a potent inhibitor of Notch-mediated transcriptional activation, achieved stable long-term reconstitution of irradiated hosts and showed a normal frequency of progenitor fractions enriched for long-term HSCs. Similar results were observed with cells lacking CSL/RBPJ, a DNA-binding factor that is required for canonical Notch signaling. Notch-deprived progenitors provided normal long-term reconstitution after secondary competitive transplantation. Furthermore, Notch target genes were expressed at low levels in primitive hematopoietic progenitors. Taken together, these results rule out an essential physiological role for cell-autonomous canonical Notch signals in HSC maintenance. PMID- 18397756 TI - N-cadherin expression level distinguishes reserved versus primed states of hematopoietic stem cells. AB - Osteoblasts expressing the homophilic adhesion molecule N-cadherin form a hematopoietic stem cell (HSC) niche. Therefore, we examined how N-cadherin expression in HSCs relates to their function. We found that bone marrow (BM) cells highly expressing N-cadherin (N-cadherin(hi)) are not stem cells, being largely devoid of a Lineage(-)Sca1(+)cKit(+) population and unable to reconstitute hematopoietic lineages in irradiated recipient mice. Instead, long term HSCs form distinct populations expressing N-cadherin at intermediate (N cadherin(int)) or low (N-cadherin(lo)) levels. The minority N-cadherin(lo) population can robustly reconstitute the hematopoietic system, express genes that may prime them to mobilize, and predominate among HSCs mobilized from BM to spleen. The larger N-cadherin(int) population performs poorly in reconstitution assays when freshly isolated but improves in response to overnight in vitro culture. Their expression profile and lower cell-cycle entry rate suggest N cadherin(int) cells are being held in reserve. Thus, differential N-cadherin expression reflects functional distinctions between two HSC subpopulations. PMID- 18397757 TI - The transcription factor EGR1 controls both the proliferation and localization of hematopoietic stem cells. AB - EGR1 is a member of the immediate early response transcription factor family and functions in cell growth, development, and stress responses in many tissues. Here we report an additional role for EGR1 in regulating homeostasis of hematopoietic stem cells (HSCs). HSCs normally express Egr1 at high levels, but dramatically downregulate its expression when induced to divide and migrate. Consistent with this finding, mice lacking Egr1 exhibit significant increases in steady-state levels of dividing HSCs in the bone marrow (BM), and a striking spontaneous mobilization of HSCs into the peripheral blood. These data identify EGR1 as a transcriptional regulator of stem cell migration that normally functions to promote HSC quiescence and retention in the niche. The ability of this single factor to regulate both proliferation and mobilization of HSCs suggests that EGR1 commands a genetic program that coordinates stem cell division and migration to maintain appropriate HSC number and function. PMID- 18397758 TI - Ontogeny and multipotency of neural crest-derived stem cells in mouse bone marrow, dorsal root ganglia, and whisker pad. AB - Although recent reports have described multipotent, self-renewing, neural crest derived stem cells (NCSCs), the NCSCs in various adult rodent tissues have not been well characterized or compared. Here we identified NCSCs in the bone marrow (BM), dorsal root ganglia, and whisker pad and prospectively isolated them from adult transgenic mice encoding neural crest-specific P0-Cre/Floxed-EGFP and Wnt1 Cre/Floxed-EGFP. Cultured EGFP-positive cells formed neurosphere-like structures that expressed NCSC genes and could differentiate into neurons, glial cells, and myofibroblasts, but the frequency of the cell types was tissue source dependent. Interestingly, we observed NCSCs in the aorta-gonad-mesonephros region, circulating blood, and liver at the embryonic stage, suggesting that NCSCs migrate through the bloodstream to the BM and providing an explanation for how neural cells are generated from the BM. The identification of NCSCs in accessible adult tissue provides a new potential source for autologous cell therapy after nerve injury or disease. PMID- 18397759 TI - Amyloid toxicity in skeletal myoblasts: Implications for inclusion-body myositis. AB - Skeletal muscle disorder, inclusion-body myositis (IBM) has been known for accumulation of amyloid characteristic proteins in muscle. To understand the biophysical basis of IBM, the interaction of amyloid fibrils with skeletal myoblast cells (SMC) has been studied in vitro. Synthetic insulin fibrils and Abeta(25-35) fibrils were used for this investigation. From the saturation binding analysis, the calculated dissociation constant (K(d)) for insulin fibril and Abeta(25-35) fibrils were 69.37+/-11.17nM and 115.60+/-12.17nM, respectively. The fibrillar insulin comparatively has higher affinity binding to SMC than Abeta fibrils. The competitive binding studies with native insulin showed that the amount of bound insulin fibril was significantly decreased due to displacement of native insulin. However, the presence of native insulin is not altered the binding of beta-amyloid fibril. The cytotoxicity of insulin amyloid intermediates was measured. The pre-fibrillar intermediates of insulin showed significant toxicity (35%) as compared to matured fibrils. Myoblast treated with beta-amyloid fibrils showed more oxidative damage than the insulin fibril. Cell differentiating action of amyloidic insulin was assayed by creatine kinase activity. The insulin fibril treated cells differentiated more slowly compared to native insulin. However, beta-amyloid fibrils do not show cell differentiation property. These findings reinforce the hypothesis that accumulation of amyloid related proteins is significant for the pathological events that could lead to muscle degeneration and weakness in IBM. PMID- 18397760 TI - Photolysis and photoisomerization of bilirubin in serum specimens exposed to room lighting. PMID- 18397761 TI - Biochemical and molecular characterization of a detergent-stable serine alkaline protease from Bacillus pumilus CBS with high catalytic efficiency. AB - We have described previously the potential use of an alkaline protease from Bacillus pumilus CBS as an effective additive in laundry detergent formulations [B. Jaouadi, S. Ellouz-Chaabouni, M. Ben Ali, E. Ben Messaoud, B. Naili, A. Dhouib, S. Bejar, A novel alkaline protease from Bacillus pumilus CBS having a high compatibility with laundry detergent and a high feather-degrading activity, Process Biochem, submitted for publication]. Here, we purified this enzyme (named SAPB) and we cloned, sequenced and over-expressed the corresponding gene. The enzyme was purified to homogeneity using salt precipitation and gel filtration HPLC. The pure protease was found to be monomeric protein with a molecular mass of 34598.19Da as determined by MALDI-TOF mass spectrometry. The NH2-terminal sequence of first 21 amino acids (aa) of the purified SAPB was AQTVPYGIPQIKAPAVHAQGY and was completely identical to proteases from other Bacillus pumilus species. This protease is strongly inhibited by PMSF and DFP, showing that it belongs to the serine proteases superfamily. Interestingly, the optimum pH is 10.6 while the optimum temperature was determined to be 65 degrees C. The enzyme was completely stable within a wide range of pH (7.0-10.6) and temperature (30-55 degrees C). One of the distinguishing properties is its catalytic efficiency (kcat/Km) calculated to be 45,265min(-1)mM(-1) and 147,000min(-1)mM(-1) using casein and AAPF as substrates, respectively, which is higher than that of Subtilisin Carlsberg, Subtilisin BPN' and Subtilisin 309 determined under the same conditions. In addition, SAPB showed remarkable stability, for 24h at 40 degrees C, in the presence of 5% Tween-80, 1% SDS, 15% urea and 10% H2O2, which comprise the common bleach-based detergent formulation. The sapB gene encoding SAPB was cloned, sequenced and over-expressed in Escherichia coli. The purified recombinant enzyme (rSAPB) has the same physicochemical and kinetic properties as the native one. SapB gene had an ORF of 1149bp encoding a protein of 383 aa organized into a signal peptide (29 aa), a pro-protein (79 aa) and a mature enzyme (275 aa). The deduced amino acid sequence inspection displays an important homology with other bacterial proteases. The highest homology of 98.1% was found with BPP-A protease from Bacillus pumilus MS 1, with only 8 aa of difference. PMID- 18397762 TI - Chloroquine mediates specific proteome oxidative damage across the erythrocytic cycle of resistant Plasmodium falciparum. AB - Resistance of Plasmodium falciparum to chloroquine hinders malaria control in endemic areas. Current hypotheses on the action mechanism of chloroquine evoke its ultimate interference with the parasite's oxidative defence systems. Through carbonyl derivatization by 2,4-dinitrophenylhydrazine and proteomics, we compared oxidatively modified proteins across the parasite's intraerythrocytic stages in untreated and transiently IC(50) chloroquine-treated cultures of the chloroquine resistant P. falciparum strain Dd2. Functional plasmodial protein groups found to be most oxidatively damaged were among those central to the parasite's physiological processes, including protein folding, proteolysis, energy metabolism, signal transduction, and pathogenesis. While an almost constant number of oxidized proteins was detected across the P. falciparum life cycle, chloroquine treatment led to increases in both the extent of protein oxidation and the number of proteins oxidized as the intraerythrocytic cycle progressed to mature stages. Our data provide new insights into early molecular effects produced by chloroquine in the parasite, as well as into the normal protein oxidation modifications along the parasite cycle. Oxidized proteins involved in the particular parasite drug-response suggest that chloroquine causes specific oxidative stress, sharing common features with eukaryotic cells. Targeting these processes might provide ways of combating chloroquine-resistance and developing new antimalarial drugs. PMID- 18397763 TI - RNA interference-mediated inhibition of hepatocyte nuclear factor 1alpha identifies target genes. AB - Hepatocyte nuclear factor 1alpha (HNF1alpha) is a homeodomain transcription factor that is central to co-ordinated differentiation of a number of cell lineages, including hepatocytes in the liver and islet cells in the pancreas. HNF1alpha interacts directly with other transcription factors and co-factors and is involved in chromatin modification to alter gene expression. To further investigate the pivotal role of HNF1alpha in transcriptional control pathways we utilized RNA interference. An siRNA oligonucleotide specific for HNF1alpha reduced HNF1alpha protein levels by up to 70% in transient transfections of Caco2 cells. The same sequence incorporated into an shRNAi reduced protein levels by up to 90% in stable transfections. Microarray analysis of RNA from cell lines with stable RNAi-mediated down-regulation of HNF1alpha, identified genes known to be regulated by this transcription factor and also novel genes. PMID- 18397764 TI - Biotechnological routes to pyruvate production. AB - Pyruvate is an important metabolite in the central metabolism of living cells. It has been widely applied in food, pharmaceutical, and agrochemical industries. Pyruvate can be produced by both chemical and biological systems. Novel biotechnological systems that can yield pyruvate have been the focus of process development in pyruvate production. In this review, we summarize recent developments related to pyruvate production by biotechnological systems, with emphasis on the enzymatic synthesis of pyruvate from the cheaper substrate lactate. PMID- 18397765 TI - Mass propagation and essential oil analysis of Artemisia vulgaris. AB - Artemisia vulgaris L. (Mugwort) is a threatened and valuable medicinal plant. Attempts have been made in this research to mass propagate its plantlets through in vitro liquid culture technology using Murashige and Skoog (MS) basal medium supplemented with 6-benzyl adenine (BA) (0.44-8.88 microM). Initially, 22.6 shoots (99.9% shooting frequency) developed from shoot tip explants cultured in MS with 4.44 microM BA at 100 ml flask capacity. This was further subcultured at increasing flask capacity (150, 250, and 500 ml) for shoot proliferation. Of the different concentrations of BA and flask capacities tested, 4.44 microM BA and 500 ml flask capacity were found to produce a maximum of 85.5 shoots after 30 d of culture. Shoot proliferation was found to increase with increasing flask capacity whereas shoot number decreased with increasing BA concentration (>4.44 microM). Individual shoots were isolated and rooted on MS medium containing 8.56 microM indole-3-acetic acid (IAA). Then the plantlets were acclimatized under standard laboratory conditions and later under greenhouse conditions. Fresh leaves were collected from greenhouse-grown plants and subjected to essential oil analysis by the simultaneous distillation and extraction method. GC-MS results revealed the presence of 88 components and the extracted oil was rich in camphor (16.8%), alpha-thujone (11.3%), germacrene D (7.2%), camphene (6.5%), 1,8-cineole (5.8%) and beta-caryophyllene (5.4%). This in vitro strategy can be a reliable method for the steady production of a large number of plants for essential oil production, which is reported for the first time for A. vulgaris. PMID- 18397766 TI - Production of extracellular bifidogenic growth stimulator (BGS) from Propionibacterium shermanii using a bioreactor system with a microfiltration module and an on-line controller for lactic acid concentration. AB - Production of a bifidogenic growth stimulator (BGS) by Propionibacterium freudenreichii subsp. shermanii (Propionibacterium shermanii) using lactic acid as a carbon source was investigated using different cultivation methods. When a continuous bioreactor system with a filtration device was used at a dilution rate of 0.075 h(-1), the average BGS concentration was 2.4 mg/l, which corresponds to a BGS productivity per cultivation time of 1.8 x 10(-1) mg x l(-1) x h(-1). The BGS productivity per cultivation time in continuous cultivation with filtration was 1.9-fold that (9.4 x 10(-2) mg x l(-1).h(-1)) in a conventional batch cultivation. In fed-batch cultivation with feed-back control using an on-line lactic acid controller with a lactic acid biosensor, it was possible to prevent substrate inhibition by maintaining the lactic acid concentration in culture broth low at 3.3 g/l, and an enhanced BGS production (31 mg/l) was successfully attained. The BGS productivity per cultivation time (2.1x10(-1) mg x l(-1) x h( 1)) in the fed-batch cultivation with feed-back control was 2.2-fold that in the conventional batch cultivation. A new bioreactor system was developed by coupling a continuous bioreactor system with a filtration device to an on-line lactic acid controller. Using the new bioreactor system, we produced BGS continuously at a high level of 47 mg/l. The BGS productivities per cultivation time (3.5 mg.l(-1) x h(-1)) and the total volume of medium used (1.7 x 10(-1) mg x l(-1) x h(-1)) obtained in the new bioreactor system were 37-fold and 2.1-fold those in the conventional batch cultivation, respectively. These results described above clearly demonstrate the positive effects of both the continuous filtration for removal of metabolites (propionic and acetic acids) inhibitory to cell growth and feed-back control of lactic acid concentration in the culture broth on BGS production by P. shermanii. This paper is the first report on BGS production by the propionic acid bacterium using lactic acid as a carbon source. PMID- 18397767 TI - Dynamics of sustainable grazing fauna and effect on performance of gas biofilter. AB - The inherent operational problems of biofilters such as a pressure drop increase and nutrient limitations were managed in a toluene-removing gas biofilter with a sustainable grazing fauna consisting of micrometazoa and ciliate protozoa. Dynamic populations of predatory nematodes (Caenorhabditis sp.), rotifers (Philodina sp.), tardigrades (Echiniscus sp.) and fly larvae represented the micrometazoa community in the filter bed. Colpoda inflata, Euplotes harpa and Acineria sp. constituted the grazing ciliate community. The spatiotemporal distribution and abundance of the grazing fauna depends on physicochemical conditions and interspecies interactions in the biofilter. Of the micro metazoa, Caenorhabditis and Philodina tolerated wide concentration ranges for toluene (0.75-2.63 g m(-3)) and CO(2) (0.92-6.08 g m(-3)) and maintained stable populations of 3.4-4.7 x 10(3) and 5.8-7.65 x 10(4) g medium(-1), respectively. The grazing fauna supported a stable toluene-degrading bacterial community composed of four Pseudomonas spp. Under a maximum toluene load of 120.72 g m(-3) h(-1), at steady-state conditions 80% toluene removal was achieved in the biofilter. Of the grazing organisms, owing to their reproductive cycle and feeding behaviour, fly larvae were not suited for application in the biofilter. Meanwhile, organisms such as nematodes, rotifers and ciliates capable of tolerating a wide pollutant concentration range and maintaining a sustainable population are ideal candidates for application in biofilter technology. PMID- 18397768 TI - Identification of HLA-A24-restricted epitopes with high affinities to Hsp70 using peptide arrays. AB - Heat shock protein 70 (Hsp70) family members are known as facilitators of immune responses by interacting with receptors on antigen-presenting cells leading to Hsp70-peptide uptake and antigen cross priming. Here, identification of human leukocyte antigen (HLA)-A24-restricted epitopes was achieved using peptide arrays for evaluation of their affinities to Hsp70 and HLA-A24 binding prediction tools. Using Hsp70 as the model antigen, the GYPVTNAVI and VFQHGKVEI peptides were identified as antigens. These peptides actually bound to HLA-A24 in the stabilization assay using T2-A*2402 cells, and induced a strong peptide-reactive cytotoxic T lymphocyte response in HLA-A24 transgenic mice after vaccination. PMID- 18397769 TI - Optimization of gellan gum production by Sphingomonas paucimobilis ATCC 31461 with nonionic surfactants using central composite design. AB - The effect of nonionic surfactants on gellan production by Sphingomonas paucimobilis was studied by the addition of 0.5, 0.75, 1.0, 1.25 and 1.5 g/l surfactants to shake flask culture. The nonionic surfactants Tween 80, Tween 40 and Triton X-100 improved gellan production by S. paucimobilis, and the maximum yield (10.44 g/l) was obtained with Triton X-100 at 0.75 g/l compared with that of the control fermentation (8.63 g/l) without surfactant. The DO profiles associated with gellan production in a 5-l laboratory fermentor showed higher oxygen and mass transfers during fermentation with surfactant than during control fermentation without surfactant. The addition of surfactant also resulted in a polymer with high viscosity as manifested by its lower acetyl content, than that obtained by control fermentation. A central composite design (CCD) was used to determine the maximum gellan production at optimum values for three process parameters (Triton X-100 concentration, pH, and temperature) each at five levels in a laboratory fermentor. The maximum gellan yield (14.62 g/l) was obtained in a 5-l laboratory fermentor with 1.0 g/l Triton X-100 and at pH 6.0 and temperature 29.6 degrees C. Further studies on the effects of agitation and DOT level demonstrated that the surfactants enhanced oxygen transfer resulting in higher gellan production (27.86 g/l) at higher agitation speed (1000 rpm) and 100% DOT level. PMID- 18397770 TI - Four-base codon-mediated saturation mutagenesis in a cell-free translation system. AB - Saturation mutagenesis is a useful technique for the structural and functional analyses of proteins and for protein engineering. However, the extensive mutagenesis of genes and expression of mutated proteins are tedious and time consuming. We have developed a simple and rapid method for the expression of mutated proteins with comprehensive single amino acid substitutions from single mutated genes having a four-base codon in a cell-free translation system. Twenty types of tRNA that were aminoacylated with one of the 20 proteinogenic amino acids and that contained a four-base anticodon were prepared by chemical aminoacylation. In the presence of one of the aminoacyl-tRNAs, a streptavidin mRNA with a four-base codon at the Tyr83 position was expressed in an Escherichia coli cell-free translation system. The N-terminus of the expressed proteins was fluorescently labeled using a fluorescent-labeled initiator Met-tRNA. Fluorescence imaging of an SDS-PAGE gel showed that all the amino acids are incorporated in response to the four-base codon; however, the incorporation efficiency was dependent on the structure of the side chains. Streptavidin mutants with comprehensive amino acid substitutions at the Tyr83, Arg84, and Tyr54 positions were used for analyzing their biotin-binding activity by dot blot analysis. These results demonstrate that this method is effective for the expression and analysis of mutated proteins with comprehensive amino acid substitutions at desired positions. PMID- 18397771 TI - Effect of light intensity and frequency of flashing light from blue light emitting diodes on astaxanthin production by Haematococcus pluvialis. AB - Flashing light from blue light emitting diodes is an effective method for the reduction of energy consumption in the bioproduction of astaxanthin by Haematococcus pluvialis. We investigated the effects of light intensity and frequency on the final astaxanthin concentration in bioproduction by H. pluvialis grown mixotrophically. The final astaxanthin concentration under illumination with flashing light, with frequencies ranging from 25 to 200 Hz, was dependent on the light intensity and on the duty cycle and was equivalent, or higher, in comparison with that under illumination with continuous light at the same incident intensity. The light intensity determined the maximum attainable concentration of astaxanthin under continuous illumination. Under illumination with flashing light, the ratio of the final astaxanthin concentration to the maximum concentration at a specific light intensity was correlated to the duty cycle in the frequency range from 25 to 200 Hz. The effect of lower frequencies on enhanced astaxanthin production under flashing light was also studied; at levels as low as 1 Hz, higher final astaxanthin concentrations were observed under flashing light compared to concentrations attained under continuous light. PMID- 18397772 TI - Effective cell separation utilizing poly(N-isopropylacrylamide)-grafted polypropylene membrane containing adsorbed antibody. AB - We previously developed a cell separation method using a poly(N isopropylacrylamide)-grafted polypropylene (PNIPAAm-g-PP) membrane containing an adsorbed monoclonal antibody (mAb). The purpose of this study is to elucidate the cell separation mechanism in detail and to design an optimal method. As the grafting yield of PNIPAAm increased, the level of the adsorption of IgG(1) and cell adhesion to the membrane decreased. After BSA was adsorbed to a PNIPAAm-g-PP membrane at 6 degrees C, where PNIPAAm was hydrophilic, a small amount of IgG(1) was adsorbed to the membrane at 37 degrees C, where PNIPAAm was hydrophobic. The desorption of the adsorbed IgG(1) was not enhanced even though temperature was reduced to 10 degrees C, where PNIPAAm was hydrophilic. These results indicate that the antibody adsorbed to the intact PP surface of the membrane predominantly contributes to the capture of target cells through the antigen-antibody reaction and that a thermoresponsive transition of PNIPAAm contributes to the detachment of the captured cells. The total number of cells recovered from a PNIPAAm-g-PP membrane containing the adsorbed mAb decreased as the grafting yield increased. A PNIPAAm-g-PP membrane with a 1.7% grafting yield containing adsorbed anti-human CD34 mAb enriched CD34-positive KG-1a cells to 85% from a 1:1 cell suspension of KG-1a cells and CD34-negative Jurkat cells. PMID- 18397773 TI - The mixed coculture effect of primary rat hepatocytes and bone marrow cells is caused by soluble factors derived from bone marrow cells. AB - Heterospheroids consisting of hepatocytes and bone marrow cells (BMCs) are formed by the mixed coculture of these cells and enhance the expression and maintenance of the liver-specific functions of hepatocytes. Not only the soluble factors derived from these cells, but also functional organoid (heterospheroid) formation, are considered to underlie this coculture effect. Therefore, in the present study, we aimed to clarify the mechanism of this co-culture effect. We performed hepatocyte monoculture with conditioned media prepared from hepatocyte cultures, BMC cultures and a coculture of hepatocytes and BMCs. When using any type of conditioned medium, no hepatocyte spheroids formed, and the hepatocytes formed a monolayer. In addition, an effect for these conditioned media was shown in terms of the albumin production and ammonia metabolism activities of the hepatocytes; conditioned medium from BMCs showed the strongest effect. The monocultured hepatocytes in the conditioned medium derived from BMCs showed equivalent albumin production and ammonia metabolism activities to the cocultured spheroids of hepatocytes and BMCs. Therefore, it was determined that the effect of the coculture of hepatocytes and BMCs was caused by soluble factors derived from BMCs. PMID- 18397774 TI - Optimization of critical medium components using response surface methodology for phenazine-1-carboxylic acid production by Pseudomonas sp. M-18Q. AB - The optimal flask-shaking batch fermentation medium for phenazine-1-carboxylic acid (PCA) production by Pseudomonas sp. M-18Q, a qscR chromosomal inactivated mutant of the strain M18 was studied using statistical experimental design and analysis. The Plackett-Burman design (PBD) was used to evaluate the effects of eight medium components on the production of PCA, which showed that glucose and soytone were the most significant ingredients (P<0.05). The steepest ascent experiment was adopted to determine the optimal region of the medium composition. The optimum composition of the fermentation medium for maximum PCA yield, as determined on the basis of a five-level two-factor central composite design (CCD), was obtained by response surface methodology (RSM). The high correlation between the predicted and observed values indicated the validity of the model. A maximum PCA yield of 1240 mg/l was obtained at 17.81 g/l glucose and 11.47 g/l soytone, and the production was increased by 65.3% compared with that using the original medium, which was at 750 mg/l. PMID- 18397775 TI - Fluorescence labeling of a cytokine with desthiobiotin-tagged fluorescent puromycin. AB - Fluorescence labeling of a cytokine at a specific site is required for observing cytokine-receptor interactions in living cells at the single-molecule level. Here, we demonstrated the C-terminus-specific fluorescence labeling of histidine tagged thrombopoietin (TPO), a ligand for Mpl, with desthiobiotin-tagged fluorescent puromycin. Fluorescent TPO, purified by tandem affinity purification, stimulated the proliferation of Mpl-expressing cells. Within 10 min of its addition, fluorescent TPO was found to be diffusely distributed on the cell membranes of Mpl-expressing cells, and gradually accumulated to form fluorescent spots. This method is applicable for studying the spatial and temporal distributions of cytokines in individual living cells. PMID- 18397776 TI - Another multiheme protein, hydroxylamine oxidoreductase, abundantly produced in an anammox bacterium besides the hydrazine-oxidizing enzyme. AB - A hydroxylamine oxidoreductase (HAO) was purified from anammox sludge in which an anammox bacterium, strain KSU-1, was dominant. The enzyme was a 118-kDa homodimer composed of a 53-kDa subunit. With phenazine methosulfate and 3-(4,5-dimethyl-2 thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide as electron acceptors, the V(max) and K(m) for hydroxylamine were determined as 9.6+/-0.2 micromol/min x mg and 33+/-2 microM, while those for hydrazine were 0.54+/-0.0 micromol/min x mg and 25+/-2 microM, respectively. The HAO had a P468 chromophore. These enzymatic properties were different from those of the hydrazine-oxidizing enzyme (HZO), a multiheme protein abundantly produced by the KSU-1 strain, but were similar to those of the HAO purified from Candidatus Brocadia anammoxidans. The hao gene exists upstream of the hzoB gene, which codes for the HZO. The sequence deduced from the hao gene indicated eight c-type heme binding motifs and showed 87% identity with a polypeptide encoded by an open reading frame (kustc1061) in the genome of an anammox bacterium Candidatus Kuenenia stuttgartiensis. These suggested that the HAO is an indispensable enzyme and well conserved in anammox bacteria, similar to the HZO. This enzyme might therefore be a specific hydroxylamine oxidoreductase for anammox bacteria. PMID- 18397777 TI - Quantitative analysis of anionic metabolites for Catharanthus roseus by capillary electrophoresis using sulfonated capillary coupled with electrospray ionization tandem mass spectrometry. AB - This paper describes a practical method to quantify anionic metabolites using contemporary capillary electrophoresis-electrospray ionization-tandem mass spectrometry (CE-ESI-MS/MS). The use of sulfonated capillary permitted a simultaneous analysis of sugar phosphates, organic acids, nucleotides and coenzyme A compounds with only one CE condition. This capillary also improved the repeatability and sensitivity. MS/MS with multiple reaction monitoring (MRM) detection was utilized to achieve significant selectivity and sensitivity. Under optimized CE-ESI-MS/MS system, the detection limits of 53 metabolites at signal to-noise ratio of 3 were between 0.040 and 8.8 mumol/l. The relative standard deviations (RSDs) for the majority anionic metabolites were better than 0.5% for migration times, and better than 10.0% for peak areas (n=6). Matrix effects by contaminants in sample solution in CE-ESI-MS/MS analysis were removed dramatically by the sample preparation method with liquid-liquid fractionation and ultrafiltration procedure. Furthermore, the developed method was successfully applied to determine anionic metabolites in a cultured cell of Catharanthus roseus. Accumulation of some metabolites including shikimate, malate, and sedoheptulose 7-phosphate by elicitation of methyl jasmonate was observed. The result would show shikimate, tricarboxylic acid and pentose phosphate pathways were activated. Our method will be useful for detailed analysis of primary metabolism dynamics. PMID- 18397778 TI - Rational screening of antibodies and design of sandwich enzyme linked immunosorbant assay on the basis of a kinetic model. AB - A rational strategy for the rapid establishment of a sensitive sandwich enzyme linked immunosorbant assay was developed. The kinetic properties required for the solid-phase and enzyme-conjugated antibodies of sandwich ELISA were determined rationally on the basis of a kinetic model describing antibody-antigen interaction. Some antibodies possessing the required kinetic properties against a model antigen, C-reactive protein (CRP), were successfully isolated from a phage antibody library under the screening conditions that were designed on the basis of simulation results. The best combination of solid-phase and enzyme-conjugated antibodies that gives the most sensitive sandwich ELISA was determined by simulation on the basis of the apparent association and dissociation rate constants of the isolated antibodies. It was confirmed by experiment that the sandwich ELISA using the best combination of antibodies was actually the most sensitive one. Our strategy would be useful for the rapid establishment of sensitive sandwich ELISAs compared with the traditional hybridoma method in which the best condition is determined by trial and error. PMID- 18397779 TI - Modified fuzzy gap statistic for estimating preferable number of clusters in fuzzy k-means clustering. AB - In clustering methods, the estimation of the optimal number of clusters is significant for subsequent analysis. Without detailed biological information on the genes involved, the evaluation of the number of clusters becomes difficult, and we have to rely on an internal measure that is based on the distribution of the data of the clustering result. The Gap statistic has been proposed as a superior method for estimating the number of clusters in crisp clustering. In this study, we proposed a modified Fuzzy Gap statistic (MFGS) and applied it to fuzzy k-means clustering. For estimating the number of clusters, fuzzy k-means clustering with the MFGS was applied to two artificial data sets with noise and to two experimentally observed gene expression data sets. For the artificial data sets, compared with other internal measures, the MFGS showed a higher performance in terms of robustness against noise for estimating the optimal number of clusters. Moreover, it could be used to estimate the optimal number of clusters in experimental data sets. It was confirmed that the proposed MFGS is a useful method for estimating the number of clusters for microarray data sets. PMID- 18397780 TI - Synthesis and degradation of acyl peptide using enzyme from Pseudomonas aeruginosa. AB - The detailed properties of the enzyme from Pseudomonas aeruginosa, which catalyzes the N-acyl linkage between myristic acid and the N-terminal glycine residue of the octapeptide GNAAAARR-NH(2) (PKA) in aqueous solution without ATP and CoA, were studied. The substrate specificity for the acyl peptide in the synthetic reaction was examined, and it was found that at least eight amino acid residues are required for the reaction and that the N-terminal glycine residue is not absolutely essential for the reaction because the activity was detected using the octapeptide that has an N-terminal alanine. The activity was also strongly affected by the amino acid sequence because the activity was very weak in the reaction using GARASVLS-NH(2) (HIV-1p17(gag)). The substrate specificity for fatty acids was also examined. In the reactions using lauric acid and decanoic acid, only slight activities were detected; however, those activities were very small compared with the activity in the reaction using myristic acid. In addition, the degradation of myristoyl PKA by the enzyme was detected, although there are only a few reports on demyristoylation. The optimum pH and temperature of the degradation reaction were consistent with those of the synthetic reaction. The degradation reaction was inhibited by divalent cations. PMID- 18397781 TI - Improved production of ginsenosides in suspension cultures of ginseng by medium replenishment strategy. AB - The objective of this study was to improve the accumulation of ginsenosides by the adventitious root cultures of ginseng, which are important secondary metabolites with pharmaceutical applications. The adventitious roots were cultured in bioreactors for 50 d using 1.5-strength Murashige and Skoog (MS) medium supplemented with 10 mg/l indole acetic acid and 75 g/l sucrose. Kinetic studies of the nutrient composition of the spent medium revealed the gradual depletion of various inorganic nutrients and sugars. Cultures were supplied with fresh nutrient medium (medium exchange or replenishment with 0.75- and 1.0 strength MS medium) after 10 and 20 d of culture initiation to fulfill the nutritional requirements of adventitious roots. Medium replenishment strategy (with 1.0-strength MS medium after 20 d of culture) significantly improved the growth of adventitious roots and the biosynthesis of ginsenosides by the adventitious roots. This work is useful for the large-scale cultivation of adventitious roots for the production of ginsenosides. PMID- 18397782 TI - Formaldehyde elimination with formaldehyde and formate oxidase in membrane of acetic acid bacteria. AB - Formaldehyde elimination was successfully carried out with Acetobacter sp. SKU 14, having strong formaldehyde-oxidizing activity in the cytoplasmic membrane. Formaldehyde was decomposed via formate to carbon dioxide by formaldehyde- and formate-oxidizing activities. A resting-cell suspension of the organism was more convenient for practical purposes than the isolated membrane fraction. In Gluconobacter suboxydans IFO 12528, formaldehyde elimination was not so prominent when compared with that in Acetobacter sp. SKU 14. PMID- 18397783 TI - Bioethanol production from xylose by recombinant Saccharomyces cerevisiae expressing xylose reductase, NADP(+)-dependent xylitol dehydrogenase, and xylulokinase. AB - We constructed a set of recombinant Saccharomyces cerevisiae strains with xylose fermenting ability. A recombinant S. cerevisiae strain D-XR/ARSdR/XK, in which protein engineered NADP(+)-dependent XDH was expressed, showed 40% increased ethanol production and 23% decrease in xylitol excretion as compared with the reference strain D-XR/XDH/XK expressing the wild-type XDH. PMID- 18397784 TI - The 5'-untranslated region of the Oryza sativa alcohol dehydrogenase gene functions as a translational enhancer in monocotyledonous plant cells. AB - The 5'-untranslated region (5'-UTR) functions as a translational enhancer in monocotyledonous plant cells is necessary to express a foreign gene efficiently. Here, we show that the 5'-UTR of the rice alcohol dehydrogenase gene contributes to efficient translation in not only dicotyledonous plant cells, but also in monocotyledonous rice cells. PMID- 18397785 TI - Toxoplasma gondii: protection against colonization of the brain and muscles in a rat model. AB - The objective was to test immune protection against the formation of Toxoplasma gondii tissue cysts in rats. It has been previously shown that 50 T. gondii tissue cysts of strain Me49 are not pathogenic for CF-1 mice, whereas 1 T. gondii tissue cyst of strain M-7741, can be lethal for mice 11-13 days after subcutaneous or oral administration. In the present study, ten rats were fed T. gondii oocysts of strain Me49 and after a further 30 days they were each orally challenged with T. gondii oocysts of strain M-7741. Thirty days after this, they were euthanased and brain and muscle samples inoculated subcutaneously or orally dosed, respectively, to mice for bioassay. None of the mice died, whereas all the mice that were inoculated with brain homogenates or were fed muscle samples from four non-immunized rats that had been inoculated with T. gondii oocysts of strain M-7741, died. These results encourage further research towards achieving vaccinal protection against the formation of T. gondii tissue cysts in meat animals and people. PMID- 18397786 TI - Plasmodium chabaudi: efficacy of artemisinin + curcumin combination treatment on a clone selected for artemisinin resistance in mice. AB - Recent studies have proposed curcumin as a potential partner for artemisinin in artemisinin combination therapies to treat malaria infections. The efficacy of curcumin alone and in combination with artemisinin was evaluated on a clone of Plasmodium chabaudi selected for artemisinin resistance in vivo. The addition of piperine as an enhancer of curcumin activity was also tested. Results indicated that curcumin, both alone and in combination with piperine had only a modest antimalarial effect and was not able to reverse the artemisinin-resistant phenotype or significantly affect growth of the tested clone when used in combination with artemisinin. This is in contrast with previous in vivo work and calls for further experimental evaluation of the antimalarial potential of curcumin. PMID- 18397787 TI - Cloning and characterization of a type 3 iodothyronine deiodinase (D3) in the liver of the chondrichtyan Chiloscyllium punctatum. AB - Thyroid hormone bioactivity is finely regulated at the cellular level by the peripheral iodothyronine deiodinases (D). The study of thyroid function in fish has been restricted mainly to teleosts, whereas the study and characterization of Ds have been overlooked in chondrichthyes. Here we report the cloning and operational characterization of both the native and the recombinant hepatic type 3 iodothyronine deiodinase in the tropical shark Chiloscyllium punctatum. Native and recombinant sD3 show identical catalytic activities: a strong preference for T3-inner-ring deiodination, a requirement for a high concentration of DTT, a sequential reaction mechanism, and resistance to PTU inhibition. The cloned cDNA contains 1298 nucleotides [excluding the poly(A) tail] and encodes a predicted protein of 259 amino acids. The triplet TGA coding for selenocysteine (Sec) is at position 123. The consensus selenocysteine insertion sequence (SECIS) was identified 228 bp upstream of the poly(A) tail and corresponds to form 2. The deduced amino acid sequence was 77% and 72% identical to other D3 cDNAs in fishes and other vertebrates, respectively. As in the case of other piscivore teleost species, shark expresses hepatic D3 through adulthood. This characteristic may be associated with the alimentary strategy in which the protection from an exogenous overload of thyroid hormones could be of physiological importance for thyroidal homeostasis. PMID- 18397788 TI - A method for the design of 3D scaffolds for high-density cell attachment and determination of optimum perfusion culture conditions. AB - The application of in vitro cultured cells in tissue engineering or drug screening, aimed at complex soft tissues such as liver, requires in vivo physiological function of the cultured cells. For this purpose, the scaffold in which cells are cultured should provide a microenvironment similar to an in vivo one with a three-dimensional extracellular matrix, a high supply capacity of O(2) and nutrients, and high cell density. In this paper, we propose a method to design (1) the geometry of the scaffold, with a surface/volume ratio optimized to allow high-density (5 x 10(7)cells/mL) cell culture and (2) culture conditions that will supply optimal quantities of oxygen and nutrients. CFD modeling of mass transport was used to determine the shear stress as well as O(2) and glucose metabolism in the scaffold (20 mm width-35 mm length) for various flow rates. Validation of the model was done through comparison with flow resistance and micro-PIV experiments. CFD analysis showed the maximum metabolic rate densities for this scaffold are 6.04 x 10(-3)mol/s/m(3) for O(2) at 0.71 mL/min and 1.91 x 10(-2)mol/s/m(3) for glucose at 0.35 mL/min. PMID- 18397789 TI - Mesh-morphing algorithms for specimen-specific finite element modeling. AB - Despite recent advances in software for meshing specimen-specific geometries, considerable effort is still often required to produce and analyze specimen specific models suitable for biomechanical analysis through finite element modeling. We hypothesize that it is possible to obtain accurate models by adapting a pre-existing geometry to represent a target specimen using morphing techniques. Here we present two algorithms for morphing, automated wrapping (AW) and manual landmarks (ML), and demonstrate their use to prepare specimen-specific models of caudal rat vertebrae. We evaluate the algorithms by measuring the distance between target and morphed geometries and by comparing response to axial loading simulated with finite element (FE) methods. First a traditional reconstruction process based on microCT was used to obtain two natural specimen specific FE models. Next, the two morphing algorithms were used to compute mappings from the surface of one model, the source, to the other, the target, and to use this mapping to morph the source mesh to produce a target mesh. The microCT images were then used to assign element-specific material properties. In AW the mappings were obtained by wrapping the source and target surfaces with an auxiliary triangulated surface. In ML, landmarks were manually placed on corresponding locations on the surfaces of both source and target. Both morphing algorithms were successful in reproducing the shape of the target vertebra with a median distance between natural and morphed models of 18.8 and 32.2 microm, respectively, for AW and ML. Whereas AW-morphing produced a surface more closely resembling that of the target, ML guaranteed correspondence of the landmark locations between source and target. Morphing preserved the quality of the mesh producing models suitable for FE simulation. Moreover, there were only minor differences between natural and morphed models in predictions of deformation, strain and stress. We therefore conclude that it is possible to use mesh-morphing techniques to produce accurate specimen-specific FE models of caudal rat vertebrae. Mesh morphing techniques provide advantages over conventional specimen specific finite element modeling by reducing the effort required to generate multiple target specimen models, facilitating intermodel comparisons through correspondence of nodes and maintenance of connectivity, and lends itself to parametric evaluation of "artificial" geometries with a focus on optimizing reconstruction. PMID- 18397790 TI - A simplified model of scar contraction. AB - The healing of wounds is a complex process and the contraction of the resulting scar can have a negative impact on the neighbouring skin. A finite element model of skin simulating the contraction of a scar and deformation of the surrounding skin is presented. The skin is represented by an orthotropic-viscoelastic constitutive law, which is validated against experimental data in the literature. A simplified experimental model of a contracting scar in real skin is also developed. The pattern and size of the wrinkles formed around the contracting scar in the finite element model compare favourably with those formed in the experimental model. The orthotropic nature of skin plays a significant role in the behaviour of skin around scars -- the wrinkles have a preferential orientation that corresponds to a direction perpendicular to the Langer's lines in the skin. The pre-stress in skin (a property that is ignored in many skin models) is shown to be an important factor in wrinkle formation around scars. The proposed model can be used to analyse the suturing and closure of wounds of various shapes. PMID- 18397791 TI - Convergent host-parasite codon usage between honeybee and bee associated viral genomes. AB - By correlating the codon usage in four insects (the honeybee, red flour beetle, mosquito and fruit fly) with six honeybee host specific viruses, we found that the codon usage patterns of the bee viruses were strongly related to that of the honeybee and only weakly related to the red flour beetle. The insects shared varying degrees of codon usage similarity which roughly follow the known phylogenetic relatedness. All of the codon usage similarity can be described by relatedness-by-descent except for the high codon usage similarity between the honeybee and honeybee associated viruses. This evidence for the convergent evolution of the honeybee viruses toward the codon usage of the honeybee suggests that small host specific viral genomes have the freedom to quickly optimize codon usage to successfully parasitize their preferred host. The codon usage co evolution of the six host specific honeybee viruses towards the codon usage of the honeybee described in this paper is the first evidence for codon usage correlation between an insect host and a single stranded RNA virus. PMID- 18397792 TI - The evolution of complex life cycles when parasite mortality is size- or time dependent. AB - In complex cycles, helminth larvae in their intermediate hosts typically grow to a fixed size. We define this cessation of growth before transmission to the next host as growth arrest at larval maturity (GALM). Where the larval parasite controls its own growth in the intermediate host, in order that growth eventually arrests, some form of size- or time-dependent increase in its death rate must apply. In contrast, the switch from growth to sexual reproduction in the definitive host can be regulated by constant (time-independent) mortality as in standard life history theory. We here develop a step-wise model for the evolution of complex helminth life cycles through trophic transmission, based on the approach of Parker et al. [2003a. Evolution of complex life cycles in helminth parasites. Nature London 425, 480-484], but which includes size- or time dependent increase in mortality rate. We assume that the growing larval parasite has two components to its death rate: (i) a constant, size- or time-independent component, and (ii) a component that increases with size or time in the intermediate host. When growth stops at larval maturity, there is a discontinuous change in mortality to a constant (time-independent) rate. This model generates the same optimal size for the parasite larva at GALM in the intermediate host whether the evolutionary approach to the complex life cycle is by adding a new host above the original definitive host (upward incorporation), or below the original definitive host (downward incorporation). We discuss some unexplored problems for cases where complex life cycles evolve through trophic transmission. PMID- 18397793 TI - A general framework for the statistical exploration of the ecological niche. AB - We propose a new statistical framework for the exploratory analysis of the ecological niche, the "General niche-environment system factor analysis" (GNESFA). The data required for this analysis are (i) a table giving the values of the environmental variables in each environment unit (EU, e.g., the patches of habitat on a vector map), (ii) a set of weights measuring the availability of the EUs to the species (e.g., the proportion of the study area covered by a given patch), and (iii) a set of utilization weights describing the use of the EUs by the focal species (e.g., the proportion of detections of the species in each patch). Each row of the table corresponds to a point in the multidimensional space defined by the environmental variables, and each point is associated with two weights. The GNESFA searches the directions in this space where the two weight distributions differ the most, choosing one distribution as the reference, and the other one as the focus. The choice of the utilization as the reference corresponds to the MADIFA (Mahalanobis distances factor analysis), which identifies the directions on which the available EUs are in average the furthest from the optimum of the niche, allowing habitat suitability modelling. The choice of the availability as the reference corresponds to the FANTER (Factor analysis of the niche, taking the environment as the reference), which identifies the directions on which the niche is the furthest from the average environment (marginality) and those on which the niche is the narrowest compared with the environment (specialization). The commonly used ENFA (Ecological niche factor analysis) is at the middle point between the MADIFA and the FANTER, considering both distributions as the reference and the focus simultaneously. When used concurrently, these three analyses allow an extensive exploration of the system. PMID- 18397794 TI - A novel descriptor of amino acids and its application in peptide QSAR. AB - A novel descriptor, vector of principal component scores (VSW) for weighted holistic invariant molecular index, was derived from the principal component analysis of a matrix of 99 weighted holistic invariant molecular indices of amino acids. The scale was then applied in three panels of peptide QSARs models constructed by partial least square (PLS). The correlative coefficient R(cum)(2) and the cross-validation correlative coefficient Q(LOO)(2) of three models were 0.861 and 0.835 for 58 angiotensin-converting enzyme inhibitors, 0.873 and 0.751 for 48 bitter tasting thresholds, 0.997 and 0.954 for 12 antimicrobial polypeptides, respectively. External validation was also performed to validate the predictive power of resulting models. Compared with other 2D or 3D descriptors, the VSW scales could better characterize structural features of peptides and provide more sound statistical models. PMID- 18397795 TI - Fitting protein-folding free energy landscape for a certain conformation to an NK fitness landscape. AB - The NK fitness landscape is a mathematical landscape model with a parameter k that governs the degree of ruggedness of the landscape. We presented a procedure to fit a given landscape to the NK fitness landscape by introducing the "apparent k-value"k(app). In this paper, we defined the protein free energy (DeltaG) landscape in amino acid sequence space, where DeltaG is the folding free energy from a random coil to a "certain conformation". Applying this landscape to our fitting procedure, we examined the statistical properties of the landscape. For calculation of a conformation energy, amino acid residues are represented by points, and interaction energies among amino acid residues are given as (1+K) body interactions, that is, an unit of interacting (1+K) residues cooperatively contribute a single energy value to the conformational energy. Our results suggest that the apparent k-value of the free energy landscape is k(app) approximately K, and that the number of possible interactions among residues is unrelated to the k(app) value. This leads to the inference that k(app) takes values about 1-3 in real landscapes, if nature adopts two-body approximately four body interaction energies. PMID- 18397796 TI - Epigallocatechin-3-O-gallate inhibits TNFalpha-induced monocyte chemotactic protein-1 production from vascular endothelial cells. AB - Monocyte chemotactic protein-1 (MCP-1) plays a pivotal role in the recruitment of monocytes and thus in the development of inflammatory cardiovascular diseases. Epigallocatechin-3-O-gallate (EGCG), the major catechin derived from green tea, has multiple beneficial effects to reduce cardiovascular disease but the effects of EGCG on vascular endothelial MCP-1 production is not known. In this study, we investigated the mechanisms by which EGCG may inhibit tumor necrosis factor-alpha (TNFalpha)-induced MCP-1 production in bovine coronary artery endothelial cells. TNFalpha increased MCP-1 production in both a concentration and time-dependent manner. Inhibitors of phosphatidylinositol-3-OH kinase (PI-3 kinase), LY294002 and wortmannin, decreased TNFalpha-induced MCP-1 production. EGCG prevented TNFalpha-mediated MCP-1 production and reduced phosphorylation of Akt (Ser473). In addition, EGCG attenuated TNFalpha mediated down-regulation of TNFalpha receptor 1 (TNFR1), but not TNFR2. In conclusion, EGCG inhibited TNFalpha-induced MCP-1 production. Moreover, EGCG inhibited Akt phosphorylation as well as TNF activation of TNFR1, which subsequently resulted in reduced MCP-1 production. These data provide a novel mechanism where the green tea flavonoid, EGCG, could provide direct vascular benefits in inflammatory cardiovascular diseases. PMID- 18397798 TI - Segregation of short-wavelength sensitive ("blue") cone signals among neurons in the lateral geniculate nucleus and striate cortex of marmosets. AB - We measured functional input from short-wavelength selective (S) cones to neurons in the dorsal lateral geniculate nucleus (LGN) and striate cortex (area V1) in anaesthetized marmosets. We found that most magnocellular (MC) and parvocellular (PC) cells receive very little (<5%) functional input from S cones, whereas blue on cells of the koniocellular (KC) pathway receive dominant input from S cones. Cells dominated by S cone input were not encountered in V1, but V1 cells received more S cone input than PC or MC cells. This suggests that S cone inputs are distributed broadly among neurons in V1. No differences in strength of S cone inputs were seen on comparing dichromatic and trichromatic marmosets, suggesting that the addition of a medium-long wavelength selective cone-opponent ("red green") channel to a dichromatic visual system does not detectably affect the chromatic properties of the S cone pathways. PMID- 18397799 TI - Persistent organochlorine pesticide residues in fish, sediments and water from Lake Bosomtwi, Ghana. AB - Some organochlorine pesticide residues in tilapia fish (Tilapiazilli), sediment and water samples from Lake Bosomtwi (the largest natural lake in Ghana) were determined to find out the extent of pesticide contamination and accumulation in the lake. The extracted residues were analyzed on a micro-capillary gas chromatograph equipped with electron capture detector. DDE (p,p'-1,1-dichloro-2,2 bis-(4-chlorophenyl)ethylene) was the predominant residue in all the samples analyzed; detected in 82% of water samples, 98% of sediment samples and 58% of fish samples at concentrations of 0.061+/-0.03 ng g(-1), 8.342+/-2.96 ng g(-1) and 5.232+/-1.30 ng g(-1), respectively. DDT (p,p'-1,1,1-trichloro-2,2-bis-(4 chlorophenyl)ethane) was detected in 78% at a mean concentration 0.012+/-0.62 ng g(-1) of water samples analyzed. The mean concentrations of DDT in sediments and fish were 4.41+/-1.54 ng g(-1) and 3.645+/-1.81 ng g(-1), respectively. The detection of lower levels of DDT than its metabolite, DDE, in the samples implies that the presence of these contaminants in the lake is as result of past usage of the pesticides. PMID- 18397800 TI - Evaluating contamination of dredges and disposal criteria in Greek coastal areas. AB - Sediments were collected from areas that were going to be dredged located at six small Greek ports, as well as from possible disposal areas. In order to assess the contamination level of dredged material, the samples were analysed for pH, organic matter, the metals Cd, Pb, Hg, Cr, Ni, Cu, Zn and major organic compounds (PAHs, PCBs). The sediments' quality was evaluated by two approaches: (i) by classifying each area according to two sets of Sediment Quality Guidelines (SQGs) and (ii) by calculating the contamination factors and degree for each area. The results showed that organic compounds were detected in trace amounts in 14% of the port samples belonging to the PAHs group. The characterization of the sediment samples was based on the metals' concentration showing that the majority of the areas to be dredged (80%) are classified as of medium-low contamination according to the SQGs. However, according to the contamination degree, only 30% of the areas are classified as of low contamination, while 40% as of moderate contamination and 30% of the areas as of considerable contamination. From the point of view of "no disturbance" of the ecosystem, decisions about dumping should be taken only after evaluating the material to be dredged. If the SQGs are exceeded, dumping must be forbidden. Otherwise, further evaluation is needed based on the comparison with the receiving area, i.e. by estimating the contamination degree. Dumping could be allowed if the contamination degree is found to be below moderate. PMID- 18397797 TI - Tx2-6 toxin of the Phoneutria nigriventer spider potentiates rat erectile function. AB - The venom of the spider Phoneutria nigriventer contains several toxins that have bioactivity in mammals and insects. Accidents involving humans are characterized by various symptoms including penile erection. Here we investigated the action of Tx2-6, a toxin purified from the P. nigriventer spider venom that causes priapism in rats and mice. Erectile function was evaluated through changes in intracavernosal pressure/mean arterial pressure ratio (ICP/MAP) during electrical stimulation of the major pelvic ganglion (MPG) of normotensive and deoxycorticosterone-acetate (DOCA)-salt hypertensive rats. Nitric oxide (NO) release was detected in cavernosum slices with fluorescent dye (DAF-FM) and confocal microscopy. The effect of Tx2-6 was also characterized after intracavernosal injection of a non-selective nitric oxide synthase (NOS) inhibitor, L-NAME. Subcutaneous or intravenous injection of Tx2-6 potentiated the elevation of ICP/MAP induced by ganglionic stimulation. L-NAME inhibited penile erection and treatment with Tx2-6 was unable to reverse this inhibition. Tx2-6 treatment induced a significant increase of NO release in cavernosum tissue. Attenuated erectile function of DOCA-salt hypertensive rats was fully restored after toxin injection. Tx2-6 enhanced erectile function in normotensive and DOCA salt hypertensive rats, via the NO pathway. Our studies suggest that Tx2-6 could be important for development of new pharmacological agents for treatment of erectile dysfunction. PMID- 18397801 TI - Effect of parental exposure to trenbolone and the brominated flame retardant BDE 47 on fertility in rainbow trout (Oncorhynchus mykiss). AB - We exposed sexually maturing male rainbow trout (Oncorhynchus mykiss) to BDE-47 (a polybrominated diphenyl ether) and female rainbow trout to trenbolone (an anabolic steroid). Male trout were orally exposed for 17 days to 55 microg/kg/day BDE-47 and female trout continuously exposed for 60-77 days to a measured trenbolone water concentration of 35 ng/L. After the exposure, eggs and semen were collected and in vitro fertilization trials performed using a sperm:egg ratio of 300,000:1. In the BDE-47 study, eggs from control females were fertilized with semen from exposed males, while in the trenbolone study, eggs from exposed females were fertilized with semen from control males. All treatments were evaluated at two-three early developmental time-points representing first cleavage (0.5 day), embryonic keel (9 days), and eyed stages (19 days), respectively. The results indicated that BDE-47 exposure did not alter fertility as embryonic survival was similar between control and exposed groups. Trenbolone exposure also did not alter embryo survival. However, in the embryos fertilized with eggs from trenbolone exposed females, a noticeable delay in developmental progress was observed. On day 19 when eye development is normally complete, the majority of the embryos either lacked eyes or displayed under developed eyes, in contrast to control embryos. This finding suggests steroidal androgen exposure in sexually maturing female rainbow trout can impact developmental timing of F1 offspring. PMID- 18397802 TI - Comet assay in phytoplankton as biomarker of genotoxic effects of environmental pollution. AB - The alkaline comet assay was tested on different microalgae: the dinoflagellates, Karenia mikimotoi and Alexandrium minutum, and the diatom, Chaetoceros gracilis. The microalgae were exposed during their exponential growth to the model direct genotoxicant, hydrogen peroxide (1h, 5 and 100muM H2O2). Following H2O2 exposure, the comet assay was validated only for K. mikimotoi for which genotoxicity was observed from the lowest tested concentration of 5 microM with a concentration dependent effect. C. gracilis was too small in size (4 microm) to be correctly analysed. For A. minutum, our lysis buffer was not strong enough to digest the cellulosic thecal plates. For K. mikimotoi, the comet assay was thus applied for the study of the genotoxic effects of different pesticides: epoxiconazole (as Opus formulation), chlorpyriphos-ethyl (as Dursban formulation) and endosulfan at 1, 10 and 100 microg of active substance/L for 24h. Exposure to epoxiconazole in formulation resulted in an increase in the extent of DNA strand breaks at the highest tested concentration icro/L. Endosulfan exposure resulted in DNA damage for K. mikimotoi nuclei. Genotoxicity was observed from 1 microg/L of endosulfan and was not concentration dependent. PMID- 18397804 TI - Effects of nanoparticles in Mytilus edulis gills and hepatopancreas - a new threat to marine life? AB - Every day new extraordinary properties of nanoparticles (a billionth of a meter) are discovered and worldwide millions are invested into nanotechnology and nanomaterials. Risks to marine organisms are still not fully understood and biomarkers to detect health effects are not implemented, yet. We used the filter feeding blue mussel as a model to analyse uptake and effects of nanoparticles from glass wool, a new absorbent material suggested for use in floating oil spill barriers. In both, gills and hepatopancreas we analysed uptake of nanomaterials by transmission electronmicroscopy (TEM) in unstained ultrathin sections over a period of up to 16 days. Lysosomal stability and lipofuscin content as general indicators of cellular pathology and oxidative stress were also measured. As portals of uptake, diffusion and endocytosis were identified resulting in nanoparticle accumulation in endocytotic vesicles, lysosomes, mitochondria and nuclei. Dramatic decrease of lysosomal membrane stability occurred after 12h of exposure. Lysosomal damage was followed by excessive lipofuscin accumulation indicative of severe oxidative stress. Increased phagocytosis by granulocytes, autophagy and finally apoptosis of epithelial cells of gills and primary and secondary digestive tubules epithelial cells indicated progressive cell death. These pathological responses are regarded as general indices of toxic cell injury and oxidative stress. By the combinational use of biomakers with the ultrastructural localisation of nanoparticle deposition, final evidence of cause effect relationships is delivered. PMID- 18397803 TI - Construction of subtracted EST and normalised cDNA libraries from liver of chemical-exposed three-spined stickleback (Gasterosteus aculeatus) containing pollutant-responsive genes as a resource for transcriptome analysis. AB - The three-spined stickleback (Gasterosteus aculeatus) is ideally suited to laboratory studies, while its wide distribution in the northern hemisphere gives it great potential as a sentinel organism. In the setting of a UK-wide collaboration (Fish Toxicogenomics) we have developed a microarray for transcriptomic analysis of chemical responses in populations of G. aculeatus under laboratory and field conditions. Although several EST libraries are available for this species none are from chemical-exposed fish and thus unlikely to include a full set of pollutant-responsive genes. To harvest such transcripts cDNA libraries were produced from liver of chemical-exposed mature males. Two normalised full-length libraries were generated by different methods: (1) partial subtraction of polyA+ RNA against solid-phase cDNA using magnetic bead technology; (2) degradation of double stranded cDNA formed by abundant transcripts. To enrich for pollutant-responsive genes a subtracted EST library was also generated. For each library approximately 1.5K clones were sequenced and characterised using Blast2GO. All libraries contained pollutant-responsive transcripts not previously available while additionally the subtracted library was generally enriched approximately 1.2-10-fold for transcripts expected to be induced in response to the pollutants. PMID- 18397805 TI - Mosquito cells accommodate balanced, persistent co-infections with a densovirus and Dengue virus. AB - To study persistent viral co-infections in arthropods, we first produced stable, persistently infected C6/36 mosquito cell cultures by serial passage of exponentially growing whole cells infected with either a densovirus (AalDNV) or Dengue virus (DEN-2). We then obtained stable, persistent co-infections by reciprocal super-challenge and similar passaging. Persistently infected cultures did not differ from naive-cell cultures in growth rate and cell morphology. Nor did they differ in high production of both viruses with high infection rates for naive C6/36 cells. Immunocytochemistry revealed that 99-100% of the cells were coinfected but that super-infection order had some effect on antigen distribution for the two viruses. Our results combined with existing field information and previously published experimental work suggest that the capacity to support stable, viral co-infections may be a general phenomenon for arthropod cells, and that they may be achieved easily and rapidly by serial passaging of whole cultured cells. Such persistent infections would facilitate studies on interactions between co-infecting viruses. PMID- 18397806 TI - Ultrastructural alterations in midgut and Malpighian tubules of Boettcherisca peregrina exposure to cadmium and copper. AB - The effects of Cu and Cd at their at their low concentrations (80microg/g diet) on the morphology and ultrastructure of the midgut and Malpighian tubules of Boettcherisca peregrina larvae were observed by light and transmission electron microscopy. After exposure to both metals, the midgut got darker, shorter, and thicker than in control, and many strumae occurred on the surface of the midgut. Similarly, Malpighian tubules got shorter and thinner. Ultrastructural alterations in the midgut included mitochondrial condensation, swelling, and lysis. The rough endoplasmic reticulum (rER) showed dilation and vesiculation. The microvilli were shortened and disorganized. The stored glycogens increased and many mineral spherites appeared along with lipid droplets decreased. Ultrastructural alterations observed in the Malpighian tubules included rER vesiculation and mitochondria swelling with loss of cristae. Shortened and disordered microvilli, increased numbers of large hydropic vacuoles, and mineral spherites were also observed. PMID- 18397807 TI - Occurrence of the transgenic corn cry1Ab gene in freshwater mussels (Elliptio complanata) near corn fields: evidence of exposure by bacterial ingestion. AB - The purpose of this study was to examine the contamination of cry1 and cry1Ab genes from Bacillus thuringiensis and transgenic corn in feral freshwater mussels collected from sites located in proximity of corn fields. In addition, mussels were transplanted for 2 months to a site in the Huron River, upstream to the Richelieu River, which is subject to intensive corn farming. Mussels were significantly contaminated by both genes in their gills, digestive glands, and gonads, as determined by qPCR methodology. Gene sequence analysis confirmed the presence of transgenic corn cry1Ab gene in mussel tissues. In an attempt to explain the presence of the transgene in mussel tissues, heterotrophic bacteria were grown from surface water and sediment samples on agar plates in the Richelieu River in May and August. The transgene was found at two out of six surface water samples and in one sediment sample. The study revealed that exposure to transgenic corn cry1Ab gene in mussels seems to proceed by ingestion of microorganisms during feeding. PMID- 18397808 TI - Toxicity and behavioral effect of 3beta,24,25-trihydroxycycloartane and beddomei lactone on the rice leaffolder Cnaphalocrocis medinalis (Guenee) (Lepidoptera: Pyralidae). AB - Treatment of Dysoxylum pure triterpenes 3beta,24,25-trihydroxycycloartane and beddomei lactone to the rice leaffolder (RLF), Cnaphalocrocis medinalis (Guenee) (Lepidoptera: Pyralidae), resulted in prolonged larval duration and reduced larval weight. In leaf cut choice assay and topical application experiments, beddomei lactone and 3beta,24,25-trihydroxycycloartaneto showed strong antifeedant and growth inhibitor activity against fourth instar larvae of C. medinalis. Also average leaf consumption was decreased (89%) by the treatment of the two terpinoids when compared with controls. Number of eggs laid by the female (fecundity) was decreased and oviposition deterrence index was increased due to the treatment. This result further shows that the pure triterpenes of Dysoxylum act as both an antifeedant and chronic toxin to the rice leaffolder larvae. PMID- 18397809 TI - The toxic effects of toluene on the optokinetic nystagmus in pigmented rats. AB - The effects of 375 mgm(-3) (100 ppm) toluene in air inhalation were evaluated on pigmented rats during either repeated exposures over five consecutive days 3h a day or during a single 4-h exposure. At the end of the inhalation period, the animals were returned to fresh air to evaluate their ability to recover optokinetic performance. The optokinetic responses were analyzed using a magnetic search coil technique previously described. After repeated toluene exposure, the eye position at rest of all the rats was unsteady. In response to visual stimulation, the eye velocity was slower and more irregular than in the control state. At the end of the stimulation, the environment of the animals became stationary, but the eye did not immediately return to a fixed stable position. A similar effect was observed after a single exposure. An increase of the optokinetic deficit was observed after single or repeated 375 mgm(-3) toluene exposures. No recovery was observed even after a single exposure. In view of the fact that toluene is a widely used solvent, these results show that inhalation of low concentrations, even for short single exposures, must be taken into account, because gaze destabilization could cause vertigo symptoms. PMID- 18397810 TI - Oxidative stress and potential biomarkers in tomato seedlings subjected to soil lead contamination. AB - Oxidative stress and defense response in leaves of tomato seedlings exposed to extraneous lead (Pb) at 0-500 mg kg(-1) soil for nearly 2 months were investigated. Superoxide radical (O(2)(-)) was quantified by electron paramagnetic resonance (EPR). Results showed that levels of O(2)(-), malondialdehyde (MDA), carbonyl group and superoxide dismutase (SOD) increased with the increase of bioavailable Pb. The O(2)(-) level was well correlated with MDA, carbonyl groups and SOD activities, suggesting that O(2)(-) might be responsible for them. Intensities in two bands of SOD isoenzymes increased along with added Pb in treatments against control, implying that multigenic expression in SOD enzymes were activated to counteract O(2)(-) stress. Heat shock protein 70 (HSP70) was induced sensitively by soil Pb, which was to alleviate oxidative damage (i.e. increased carbonyl groups). The overall results indicated that HSP70 and O(2)(-) were the most sensitive parameters and the combination of them might be potential biomarkers of soil Pb contamination in tomato seedlings. PMID- 18397811 TI - In situ bioassay with Eisenia andrei to assess soil toxicity in an abandoned uranium mine. AB - The goal of this study was to develop an in situ bioassay with Eisenia andrei, deploying it in several locations of an abandoned mining area. Our objectives were two-fold: (i) we intended to validate the in situ soil bioassay procedures, while (ii) providing ecologically relevant data to complement the ongoing risk evaluation based on laboratorial assays. To promote cost- and time-effectiveness, the in situ exposure was short (48 h) and the endpoints analysed included oxidative stress biomarkers and metal content in soil and organisms. The bioassay was carried out under different experimental conditions, simulating local (natural soil) vs. control conditions (LUFA soil), and irrigation with artificial rainwater vs. irrigation with diluted acidic effluent. Variation in the data was mostly due to soil type, rather than irrigation water, and substantial spatial heterogeneity was observed. Oxidative stress biomarkers did not fully work as sensitive parameters to environmental contamination. Earthworm metal burdens suggested a potential concern in terms of bioaccumulation of some metallic elements. PMID- 18397812 TI - Isolation, purification and characterization of hemerythrin from Methylococcus capsulatus (Bath). AB - Earlier work from our laboratory has indicated that a hemerythrin-like protein was over-produced together with the particulate methane monooxygenase (pMMO) when Methylococcus capsulatus (Bath) was grown under high copper concentrations. A homologue of hemerythrin had not previously been found in any prokaryote. To confirm its identity as a hemerythrin, we have isolated and purified this protein by ion-exchange, gel-filtration and hydrophobic interaction chromatography, and characterized it by mass spectrometry, UV-visible, CD, EPR and resonance Raman spectroscopy. On the basis of biophysical and multiple sequence alignment analysis, the protein isolated from M. capsulatus (Bath) is in accord with hemerythrins previously reported from higher organisms. Determination of the Fe content in conjunction with molecular-weight estimation and mass analysis indicates that the native hemerythrin in M. capsulatus (Bath) is a monomer with molecular mass 14.8 kDa, in contrast to hemerythrins from other eukaryotic organisms, where they typically exist as a tetramer or higher oligomers. PMID- 18397813 TI - Copper(II) complexes of Neobelliera Bullata Trypsin Modulating Oostatic Factor and its analogues. AB - The stoichiometry, stability constants and solution structure of the complexes formed in the reaction of copper(II) with hexapeptide NPTNLH, i.e. the Neobelliera Bullata Trypsin Modulating Oostatic Factor (Neb-TMOF), and its analogues DPTNLH, Ac-NPTNLH and Ac-DPTNLH have been determined by potentiometric, UV-visible, CD and EPR spectroscopic methods. Upon raising pH for Ac-NPTNLH and Ac-DPTNLH peptides, copper(II) coordination starts from the imidazole nitrogen of the His(6); afterwards three deprotonated amide nitrogens are progressively involved in metal ions coordination. In a wide pH range of 4.5-8.5 for the NPTNLH and DPTNLH ligands the CuL complex dominates with the imidazole nitrogen of His(6) coordinated to form a macrochelate. The N-terminal amino group of the NPTNLH and DPTNLH peptides takes part in the coordination of the metal ion in the CuL, CuH(-1)L and CuH(-2)L complexes. However, at pH above 9 the CuH(-3)L complex with the {N(Im),3N(-)} coordination mode is formed. For the CuH(-2)L complex the spectroscopic data clearly indicate the 4N {NH(2), CO or COO(-), 2N(-), N(Im)} bonding mode with the axial coordination of the N-terminal amine group to the metal ion. PMID- 18397814 TI - Characteristic differences in the X-ray photoelectron spectrum between B-DNA and M-DNA monolayers on gold. AB - Duplex DNA monolayers were self-assembled on gold through a disulfide linkage and both B- and M-DNA conformations were studied using X-ray photoelectron spectroscopy (XPS). The film thickness, density, elemental composition and ratios for samples were analyzed and compared. The DNA surface coverage, calculated from both XPS and electrochemical measurements, was approximately 1.2 x 10(13)molecules/cm(2) for B-DNA. All samples showed distinct peaks for C 1s, O 1s, N 1s, P 2p and S 2p as expected for a thiol-linked DNA. On addition of Zn(2+) to form M-DNA the C 1s, P 2p and S 2p showed only small changes while both the N 1s and O 1s spectra changed considerably. This result is consistent with Zn(2+) interacting with oxygen on the phosphate backbone as well as replacing the imino protons of thymine (T) and guanine (G) in M-DNA. Analysis of the Zn 2p spectra also demonstrated that the concentration of Zn(2+) present under M-DNA conditions is consistent with Zn(2+) binding to both the phosphate backbone as well as replacing the imino protons of T or G in each base pair. After the M-DNA monolayer is washed with a buffer containing only Na(+) the Zn(2+) bound to the phosphate backbone is removed while the Zn(2+) bound internally still remains. PMID- 18397815 TI - Regulatory signals for endothelial podosome formation. AB - Podosomes are punctate actin-rich adhesion structures which spontaneously form in cells of the myelomonocytic lineage. Their formation is dependent on Src and RhoGTPases. Recently, podosomes have also been described in vascular cells. These podosomes differ from the former by the fact that they are inducible. In endothelial cells, such a signal can be provided by either constitutively active Cdc42, the PKC activator PMA or TGFbeta, depending on the model. Consequently, other regulatory pathways have been reported to contribute to podosome formation. To get more insight into the mechanisms by which podosomes form in endothelial cells, we have explored the respective contribution of signal transducers such as Cdc42-related GTPases, Smads and PKCs in three endothelial cell models. Results presented demonstrate that, in addition to Cdc42, TC10 and TCL GTPases can also promote podosome formation in endothelial cells. We also show that PKCalpha can be either necessary or entirely dispensable, depending on the cell model. In contrast, PKCdelta is essential for podosome formation in endothelial cells but not smooth muscle cells. Finally, although podosomes vary very little in their molecular composition, the signalling pathways involved in their assembly appear very diverse. PMID- 18397816 TI - An experimental study on collagen content and biomechanical properties of sclera after posterior sclera reinforcement. AB - BACKGROUND: The development of pathological myopia is associated with reduced scleral collagen accumulation, scleral thinning, and loss of scleral tissue, in both humans and animal models. Posterior scleral reinforcement (PSR) was considered as an effective way for treating pathological myopia. Yet it is not well understood the possible role of collagen on the sclera reinforcement mechanisms in the PSR surgery. METHODS: PSR surgery was performed on the normal adult New Zealand white rabbits eyes. Human sclera was used as reinforcement materials. At 1, 2, 3, 6, 9 months after the PSR surgery, scleral hydroxyproline (Hyp) synthesis and collagen fibers arrangement were determined by enzymolysic hydrolysis assay and histological morphology technique. An Instron test machine was used to investigate the elastic modulus of sclera. FINDINGS: It was found that the elastic modulus and Hyp content of reinforced sclera were lower at first month after surgery, and then gradually up to physiological level in the following months. Those two indexes were close to that of the normal control groups at 9 months. INTERPRETATION: These findings indicate that sclera elastic modulus was associated with both change of Hyp content and collagen fibers arrangement after PSR. The therapeutic effect of PSR surgery was confirmed not only from biological but also biomechanical aspects. PMID- 18397817 TI - Apparent lack of pharmacological effect of steviol glycosides used as sweeteners in humans. A pilot study of repeated exposures in some normotensive and hypotensive individuals and in Type 1 and Type 2 diabetics. AB - Steviol glycosides, isolated from the plant Stevia rebaudiana (Bertoni) Bertoni, have been used as safe sweetening agents for more than 30 years. Beneficial effects of high doses of steviol glycosides on hyperglycemia and hypertension have been previously described when these abnormalities are present. This study was designed to evaluate the effects of steviol glycosides on blood glucose and on blood pressure (BP) in 3 groups of individuals. This was a randomized, double blind, placebo-controlled, long-term study in three groups of patients: Group 1: subjects with Type 1 diabetes; Group 2: subjects with Type 2 diabetes; and Group 3: subjects without diabetes and with normal/low-normal BP levels. The subjects in each group were randomly allocated to active treatment (the steviol glycoside stevioside: 250mg t.d.s.) or to placebo treatment and followed-up for 3 months. Post-treatment systolic BP, diastolic BP, glucose and glycated hemoglobin (HbA1c) were not significantly different from baseline measurements, except for the placebo Type 1 diabetics group where a significant difference was observed for systolic BP and glucose. No side effects were observed in the two treatment groups. This study shows that oral steviol glycosides, taken as sweetener are well tolerated and have no pharmacological effect. PMID- 18397818 TI - On the evolutionary origins of executive functions. AB - In this paper it is proposed that the prefrontal lobe participates in two closely related but different executive function abilities: (1) "metacognitive executive functions": problem solving, planning, concept formation, strategy development and implementation, controlling attention, working memory, and the like; that is, executive functions as they are usually understood in contemporary neuroscience; and (2) "emotional/motivational executive functions": coordinating cognition and emotion/motivation (that is, fulfilling biological needs according to some existing conditions). The first one depends on the dorsolateral prefrontal areas, whereas the second one is associated with orbitofrontal and medial frontal areas. Current tests of executive functions basically tap the first ability (metacognitive). Solving everyday problems (functional application of executive functions), however, mostly requires the second ability (emotional/motivational); therefore, these tests have limited ecological validity. Contrary to the traditional points of view, recent evidence suggests that the human prefrontal lobe is similar to other primates and hominids. Other primates and hominids may possess the second (emotional executive functions) prefrontal ability, -but not the first (metacognitive executive functions) one. It is argued that metacognitive executive functions are significantly dependent on culture and cultural instruments. They probably are the result of the development and evolution of some "conceptualization instruments"; language (and written language as an extension of oral language) may represent the most important one. The second executive function ability (emotional/motivational) probably is the result of a biological evolution shared by other primates. PMID- 18397819 TI - An aqueous extract of Salacia oblonga root, a herb-derived peroxisome proliferator-activated receptor-alpha activator, by oral gavage over 28 days induces gender-dependent hepatic hypertrophy in rats. AB - Activation of peroxisome proliferator-activated receptor (PPAR)-alpha by natural and synthetic chemicals induces hepatic hypertrophy. An aqueous extract of Salacia oblonga root (SOW) is an Ayurvedic medicine with anti-diabetic and anti obesity properties. In the present study, it was found that SOW (100, 300 and 900mg/kg, once daily by oral gavage over a 28 day period) elicited dose-related increases in liver weight (LW) by 1.6%, 13.4% and 42.5%, respectively, and in the ratio of LW to body weight by 8.8%, 16.7% and 40.2%, respectively, in male rats. These effects were less pronounced in females. SOW selectively increased liver mass in male rats but Sudan red staining was not different, which indicates that hepatic lipid accumulation was similar in both genders. However, SOW even at the highest dosage did not influence serum ALT and AST activities in male or female rats. Moreover, SOW was found to activate PPAR-alpha in human hepatoma-derived HepG2 cells, as evidenced by the upregulation of PPAR-alpha and acyl-CoA oxidase mRNA expression. Thus, SOW-dependent PPAR-alpha activation may precede the development of the gender difference in hepatic hypertrophy; this process may be influenced by sex hormone status. PMID- 18397820 TI - Synergistic anti-tumor effect of combination radio- and immunotherapy by electro gene therapy plus intra-tumor injection of dendritic cells. AB - Interleukin-2 (IL-2) and granulocyte-macrophage colony stimulating factor (GM CSF) facilitate the maturation and functioning of injected DC. We developed a method of in situ electroporation using IL-2 and GM-CSF genes (EGT/cytokines), followed by intra-tumoral (i.t.) immature DC to determine the immune response at the tumor site using prostate-specific antigen-transfected CT26 cells. Three cycles of EGT/cytokines and i.t. DC inhibited tumor growth most effectively, but not superior to EGT/cytokines alone. However, the role of i.t. DC became significant when radiation was given after immunotherapy, which may have clinical implications on achieving better local control and prevention of systemic relapse. PMID- 18397821 TI - Impact of extravascular lung water index on outcomes of severe sepsis patients in a medical intensive care unit. AB - BACKGROUND: This study was designated to investigate whether extravascular lung water index (EVLI) is an independent predictor for mortality outcome in patients with severe sepsis. METHODS: This study prospectively recruited patients with severe sepsis from a medical intensive care unit (ICU) at a university affiliated hospital. In each patient, transpulmonary thermodilution was used to measure cardiovascular hemodynamics and EVLI via an arterial catheter placed within 48h of the patient meeting the criteria for severe sepsis. RESULTS: In total, 33 patients were studied. EVLI, Acute Physiology and Chronic Health Evaluation (APACHE) II score, development of acute respiratory distress syndrome, chest X ray score, lung injury score, body mass index, prior 24h fluid balance, albumin, and white blood cell counts were shown to be predictors of in-hospital survival by a bivariate analysis. In multinominal logistic regression, EVLI (adjusted odds ratio, 6.21; p=0.01; 95% confidence interval, 1.05-1.44) acted as an independent predictor for in-hospital survival. A cut-off value for EVLI of 10ml/kg had good sensitivity (88.2%) and specificity (68.7%) by ROC curve analysis. Medical ICU patients with extremely severe sepsis and a high EVLI (> or =10ml/kg) had lower in-hospital survival rate than those with a low EVLI (<10ml/kg) (15% vs. 67.7%, respectively, p<0.001. CONCLUSIONS: This investigation suggested that EVLI was an independent predictor for in-hospital survival in medical ICU patients with severe sepsis. Measurement of EVLI may be used for risk stratification among those patients. PMID- 18397822 TI - Enhanced solid waste management by understanding the effects of gender, income, marital status, and religious convictions on attitudes and practices related to street littering in Nablus - Palestinian territory. AB - Litter is recognized as a form of street pollution and a key issue for solid waste managers. Nablus district (West Bank, Palestinian Territory), which has an established network of urban and rural roads, suffers from a wide-spread litter problem that is associated with these roads and is growing steadily with a well felt negative impact on public health and the environment. The purpose of this research was to study the effects of four socio-economic characteristics (gender, income, marital status, and religious convictions) of district residents on their attitudes, practices, and behavior regarding street litter generation and to suggest possible remedial actions. All four characteristics were found to have strong correlations, not only with littering behavior and practices, but also with potential litter prevention strategies. In particular, the impact of religious convictions of the respondents on their littering habits and attitudes was very clear and interesting to observe. PMID- 18397823 TI - MUC4 interacts with ErbB2 in human gallbladder carcinoma: potential pathobiological implications. AB - Muc4 interacts with erbB2 and potentiates tumourigenesis and/or tumour growth. The expression of MUC4, the interaction of MUC4 with erbB2 and the status of erbB2 signalling in human gallbladder carcinomas were determined in order to gain a better understanding of the pathobiology. The expression levels of MUC4 protein and mRNA were increased in specimens of gallbladder carcinoma. Immunoprecipitation experiments showed an interaction between MUC4 and erbB2. This interaction was associated with the hyperphosphorylation of erbB2, MAPK and Akt and with the overexpression of cyclooxygenase-2. MUC4 was detected on the apical surface of cancerous epithelia and partially co-localised there with erbB2. Transfection experiments showed that MUC4 amplifies cell proliferation in the presence of heregulin through potentiating phosphorylation of erbB2 and its downstream signalling pathways. These findings suggest that MUC4 is up-regulated and interacts with erbB2 in human gallbladder carcinoma, and thereby support the potential implication of MUC4 in erbB2 activation. PMID- 18397824 TI - Concentration and microsatellite status of plasma DNA for monitoring patients with renal carcinoma. AB - We verified the feasibility of plasma bound method for detecting renal cell carcinoma (RCC) combining the study of plasma DNA concentration and microsatellite alterations (LOH). Plasma DNA concentration was evaluated with real-time PCR in 54 patients with renal neoplasm before surgery and in 20 of these patients during a 26-64 month follow-up. Microsatellite study was performed on tumour tissue DNA of 33 RCC clear cell (RCCcc) and on plasma DNA of 14 RCCcc patients during preoperative and/or follow-up period. Patients had a significantly high (26.4+/-48.3 ng/ml versus controls 3.2+/-1.5 ng/ml; p=0.003) preoperative plasma DNA concentration that decreased after nephrectomy. During follow-up, plasma DNA increased in 12 patients without evidence of neoplasia; 3 patients successively relapsed. Tumour tissue DNA of 25 RCCcc patients (75.8%) displayed microsatellite LOH. Preoperative plasma DNA of 9 patients harboured LOH in 5 cases (55.6%). Augmented plasma DNA of 7 patients displayed LOH in 3 cases (42.9%) at follow-up, and in 1 case preceded the recurrence of disease. Plasma DNA concentration combined with microsatellite LOH in plasma DNA may predict disease recurrence in RCC patients. PMID- 18397825 TI - Enhanced hyaluronic acid production by a two-stage culture strategy based on the modeling of batch and fed-batch cultivation of Streptococcus zooepidemicus. AB - This study aimed to enhance hyaluronic acid (HA) production by a two-stage culture strategy based on the modeling of batch and fed-batch culture of Streptococcus zooepidemicus. Batch culture had higher specific HA synthesis rate while fed-batch culture had higher specific cell growth rate. The lower specific HA synthesis rate in fed-batch culture resulted from the competition of cell growth for the common precursors at a low substrate concentration. Based on the modeling of batch and fed-batch culture of S. zooepidemicus, a two-stage culture strategy was proposed to enhance HA production. S. zooepidemicus were cultured in a fed-batch mode with sucrose concentration maintained at 1.0+/-0.2g/L during 0 8h and then batch culture was performed during 8-20h with an initial sucrose concentration of 15g/L. With the proposed two-stage culture strategy, HA production was increased to 6.6g/L compared with 5.0g/L in batch culture with the same total sucrose. The enhanced HA production by the proposed two-stage culture strategy resulted from the decreased inhibition of cell growth and the increased transformation rate of sucrose to HA. PMID- 18397826 TI - Evaluation of antiangiogenic activity of azumamides by the in vitro vascular organization model using mouse induced pluripotent stem (iPS) cells. AB - Evaluation of antiangiogenic activity of marine sponge derived azumamides by the in vitro vascular organization model using mouse induced pluripotent stem (iPS) cells was carried out. Azumamide E (5) strongly inhibited in vitro angiogenesis from iPS cells at 1.9microM while azumamide A (1) showed only weak inhibition at 19microM. These results were well correlated with HDAC inhibitory activity of these compounds, revealing the prospect of azumamides as the probe molecules useful for stem cell chemical biology. PMID- 18397827 TI - Synthesis and structure-activity relationship of histone deacetylase (HDAC) inhibitors with triazole-linked cap group. AB - Histone deacetylase (HDAC) inhibition is a recent, clinically validated therapeutic strategy for cancer treatment. Small molecule HDAC inhibitors identified so far fall in to three distinct structural motifs: the zinc-binding group (ZBG), a hydrophobic linker, and a recognition cap group. Here we report the suitability of a 1,2,3-triazole ring as a surface recognition cap group linking moiety in suberoylanilide hydroxamic acid-like (SAHA-like) HDAC inhibitors. Using "click" chemistry (Huisgen cycloaddition reaction), several triazole-linked SAHA-like hydroxamates were synthesized. Structure-activity relationship revealed that the position of the triazole moiety as well as the identity of the cap group markedly affected the in vitro HDAC inhibition and cell growth inhibitory activities of this class of compounds. PMID- 18397828 TI - Synthesis and biochemical application of 2'-O-methyl-3'-thioguanosine as a probe to explore group I intron catalysis. AB - Oligonucleotides containing 3'-S-phosphorothiolate linkages provide valuable analogues for exploring the catalytic mechanisms of enzymes and ribozymes, both to identify catalytic metal ions and to probe hydrogen-bonding interactions. Here, we have synthesized 2'-O-methyl-3'-thioguanosine to test a possible hydrogen-bonding interaction in the Tetrahymena ribozyme reaction. We developed an efficient method for the synthesis of 2'-O-methyl-3'-thioguanosine phosphoramidite in eight steps starting from 2'-O-methyl-N(2)-(isobutyryl) guanosine with 10.4% overall yield. Following incorporation into oligonucleotides using solid-phase synthesis, we used this new analogue to investigate whether the 3'-oxygen of the guanosine cofactor in the Tetrahymena ribozyme reaction serves as an acceptor for the hydrogen bond donated by the adjacent 2'-hydroxyl group. We show that regardless of whether the guanosine cofactor bears a 3'-oxygen or 3' sulfur leaving group, replacing the adjacent 2'-hydroxyl group with a 2'-methoxy group incurs the same energetic penalty, providing evidence against an interaction. These results indicate that the hydrogen bond donated by the guanosine 2'-hydroxyl group contributes to catalytic function in a manner distinct from the U(-1) 2'-hydroxyl group. PMID- 18397829 TI - 1-Aryl-2-dimethylaminomethyl-2-propen-1-one hydrochlorides and related adducts: A quest for selective cytotoxicity for malignant cells. AB - The primary objective of this study was to discover one or more clusters of compounds which are not equitoxic but display cytoselectivity toward different malignant cells. Furthermore a most important consideration is that such molecules should also display greater cytotoxic potencies to tumors than normal tissues. Two series of compounds are described which meet these criteria, namely the 1-aryl-2-dimethylaminomethyl-2-propen-1-one hydrochlorides 1a-e and 1-aryl-3 dimethylamino-2-hydroxymethyl-1-propanone hydrochlorides 2a-e. A number of these compounds possess marked cytotoxic potencies (IC(50) and CC(50) values within the 10(-6) and 10(-7) molar range) which are greater than these of the reference drug melphalan. Statistical analyses demonstrated that cytotoxic potencies are influenced by the size of the aryl substituents in series 1 and to some extent by the electronic properties of the aryl groups in series 2. The mode of action of a representative compound 1e in HL-60 cells included inducing apoptosis and activation of caspases -3, -8, and -9. PMID- 18397830 TI - Experimental characterization and Monte Carlo simulation of Si(Li) detector efficiency by radioactive sources and PIXE. AB - This work relates to the study and characterization of the response function of an X-ray spectrometry system. The intrinsic efficiency of a Si(Li) detector has been simulated with the Monte Carlo codes MCNP and GEANT4 in the photon energy range of 2.6-59.5keV. After finding it necessary to take a radiograph of the detector inside its cryostat to learn the correct dimensions, agreement within 10% between the simulations and experimental measurements with several point-like sources and PIXE results was obtained. PMID- 18397831 TI - IL-12 serum levels in patients with type 2 diabetes treated with sulphonylureas. AB - Interleukin-12 (IL-12) has been identified as a pro-inflammatory cytokine which is thought to contribute to the development of atherosclerosis. However, to date, the various associations between factors related to the course of type 2 diabetes, like metabolic compensation, beta cell secretory dysfunction, insulin resistance and IL-12 serum levels, remain unclear. Our study involved 41 patients with type 2 diabetes, 19 patients with coronary artery disease (CAD), and 19 healthy controls. We measured serum levels of fasting glucose, HbA(1)c, 1,5 anhydro-d-glucitol, and lipids. In addition, serum levels of C-peptide, insulin, proinsulin and IL-12 were assayed. HOMA(IR) score was calculated. The serum concentrations of IL-12 were higher in diabetics than in either patients with CAD or healthy controls, and were correlated with BMI, C-peptide, insulin, HOMA(IR), proinsulin and HDL serum levels. Multiple regression analysis revealed that the IL-12 serum level in type 2 diabetics primarily is dependent upon fasting proinsulin concentration. Our results demonstrate that elevated IL-12 serum levels in type 2 diabetics treated with sulphonylureas are induced especially by peripheral insulin resistance and beta cells dysfunction, as expressed by fasting serum proinsulin levels. This finding gives us hope that treatment to decrease peripheral insulin resistance and to avoid excessive proinsulin secretion might be successful in the prevention of IL-12-induced atherosclerosis. PMID- 18397832 TI - Inhibitory effect of carnosine on interleukin-8 production in intestinal epithelial cells through translational regulation. AB - The enhanced intestinal production of pro-inflammatory cytokines leads to inflammation and carcinogenesis, and therefore its down-regulation by nutrients could represent a promising therapeutic approach. We found for the first time that the secretion of interleukin-8 (IL-8) in intestinal epithelial cells stimulated by hydrogen peroxide or TNF-alpha was suppressed in the presence of carnosine (beta-Ala-His), a dietary dipeptide. Interestingly, carnosine had no influence on the stimulus-induced IL-8 mRNA expression, although the intracellular production and secretion of IL-8 were significantly inhibited by carnosine. The inhibitory effect of carnosine on the IL-8 secretion differed from that of other histidine-containing dipeptides like Gly-His, Ala-His, and anserine (beta-Ala-1-methyl-His), which inhibited both the hydrogen peroxide-induced secretion and mRNA expression of IL-8. These observations indicate that carnosine inhibited IL-8 secretion along a unique pathway, in which IL-8 production was suppressed at a post-transcriptional level, for instance, translation. The hypothesis that carnosine inhibited the translation of IL-8 mRNA is supported by the finding that the phosphorylation of eIF4E, an initiation factor, in stimulated Caco-2 cells was inhibited by carnosine. These results suggest that carnosine is a novel type of anti-inflammatory agent that down-regulates the inflammatory response in intestinal epithelial cells by a unique mechanism. PMID- 18397833 TI - Affinity-based isolation of a bacterial lipase through steric chaperone interactions. AB - Lipases are important as additives in detergent formulations but their biocatalytic potential is increasingly exploited in the synthesis of high-added value chemicals, in fine-chemical production and in the pharmaceutical industry. Traditionally, conventional purification schemes comprise several chromatographic steps. Here we report a new purification procedure of the lipase (LipA) that is endogenously secreted by the Gram-negative bacterium Burkholderia glumae. This affinity purification combines the specific binding scaffold of a lipase-specific foldase (Lif) and the intrinsic resistance to chemical denaturation of LipA. The newly devised method is less labor-intensive, is fast, leads to a homogeneous preparation and can be easily scaled up. The novel and the conventional purification strategies were evaluated in parallel and characteristics of the B. glumae lipase were analyzed via CD spectroscopy. Lipopolysaccharide (LPS) was still present in the samples purified via the conventional purification scheme and was shown to increase the thermostability of the lipase. PMID- 18397834 TI - One-step affinity purification of recombinant TATA binding proteins utilizing a modular protein interaction partner. AB - We describe a rapid and effective procedure for purifying recombinant eukaryotic TATA binding protein (TBP) from Escherichia coli. The method employs an affinity ligand comprising glutathione-S-transferase fused to the carboxyl-terminal activation domain of the transcriptional activator VP16 and an amino-terminal domain (TAND2) of the yeast TBP-associated factor TAF1. TBP can be purified without the need for extrinsic affinity tags, subsequent proteolysis, or downstream clean-up steps. This TBP purification process is rapid (requiring about 4h after bacterial harvest) and does not require sophisticated chromatographic equipment. The resulting material is monodisperse, structured, and functionally active. We demonstrate the efficacy of this method for purifying recombinant full-length or TBP core fragments encoded by yeast, humans and Arabidopsis. PMID- 18397835 TI - Lymphocyte blastogenic response to ovalbumin in a model for canine allergy. AB - Lymphocyte stimulation tests (LST) were performed in five dogs sensitised with ovalbumin (OVA) and seven healthy dogs. In addition, all five OVA-sensitised and two control dogs were tested after two in vivo provocations with OVA-containing eye drops. The isolated cells were suspended in culture media containing OVA and were cultured for up to 12 days. Proliferation was measured as reduction in 5,6 carboxylfluorescein diacetate succinimidyl ester (CFSE) intensity by flow cytometry on days 0, 3, 6, 9 and 12. A cell proliferation index (CPI) for each day and the area under the curve (AUC) of the CPI was calculated for each dog. All OVA-sensitised dogs demonstrated increased erythema after conjunctival OVA application. The presence of OVA-specific lymphocytes was demonstrated in 2/5 OVA sensitised dogs before and 4/5 after in vivo provocation. Using the AUC, the difference between OVA-sensitised and control dogs was significant in all three LST before in vivo provocation (P<0.05) and borderline significant (P=0.053) in 2/3 LST after provocation. The most significant difference in CPI was observed after 9 days of culture (P=0.001). This pilot study indicates that the LST allows detection of rare antigen specific memory T-cells in dogs previously sensitised to, but not concurrently undergoing challenge by a specific antigen. PMID- 18397836 TI - New aspects in the pathogenesis of abomasal displacement. AB - Impaired abomasal motility and an increased accumulation of gas are prerequisites for displacement of the abomasum in the cow. Predisposing factors are the breed (e.g. Holstein-Friesian, Simmental-Red-Holstein cross breeds and Guernsey), genetic background, twin pregnancy, first weeks of lactation, metabolic disorders (ketosis, increased lipomobilisation, insulin resistance), high-concentrate and low-fibre diets, as well as other concomitant diseases, such as endometritis, mastitis and claw disorders. There does not appear to be a strong correlation between increased milk yield or endotoxaemia and abomasal displacement. Recent studies have focused on possible functional disorders of the enteric nervous system within the abomasal wall, since cattle with abomasal displacement have an increased activity of neuronal nitric oxide synthase, as well as decreased acetylcholine sensitivity. In addition, there appear to be significant differences between breeds in the levels of the neurotransmitters substance P (SP) and vasoactive intestinal peptide (VIP) in the abomasal wall. For example, SP (stimulatory) was significantly less in German Holsteins in comparison to the German Fleckvieh, whereas VIP (inhibitory) was markedly increased. These risk factors may explain why Holstein cows are more susceptible to abomasal displacement than other breeds. PMID- 18397838 TI - An atom-efficient and practical synthesis of new pyridinium ionic liquids and application in Morita-Baylis-Hillman reaction. AB - New ionic liquids containing (3-chloro-2-hydroxypropyl)-functionalized pyridinium cations have been synthesized by the ultrasound-assisted, atom-efficient, room temperature reaction of pyridine with acid and 3-chloro-propylene oxide, the acid providing the anionic component of the resultant ionic liquids, and under the ultrasound, a clear yield increase results and a dramatic reduction of the reaction time accompanied by an improved quality of the products occurs. Furthermore, the application of new ionic liquids were tested as solvents in Morita-Baylis-Hillman reaction, in some cases, good results were obtained. PMID- 18397837 TI - A novel missense mutation (G43S) in the switch I region of Rab27A causing Griscelli syndrome. AB - The autosomal recessive Griscelli syndrome type II (GSII) is caused by mutations in the RAB27A gene. Typical clinical features include immunological impairment, silver-gray scalp hair, eyelashes and eyebrows and hypomelanosis of the skin. Rabs help determine the specificity of membrane trafficking steps within cells. In melanocytes, the GTP-bound form of Rab27A associates with the membranes of mature fully-pigmented melanosomes through its geranylgeranyl group. Once attached, Rab27A recruits the downstream effector Melanophilin (Mlph) and the actin-dependent motor protein Myosin Va (MyoVa). The molecular Rab27A/Mlph/MyoVA tripartite complex, which links melanosomes to the peripheral actin network, is required to achieve melanosome transfer to surrounding keratinocytes in the epidermis. Here we report a novel homozygous missense mutation c.127G>A, p.G43S in exon 2 of the RAB27A gene of an Afghani GSII patient. Laser scanning confocal microscopy showed that the G43S mutation, which is located in the highly conserved switch I region of Rab27A, induces perinuclear localization of melanosomes in normal melanocytes, and fails to restore melanosomes to the actin rich periphery in GSII melanocytes. Co-immunoprecipitation studies showed that Rab27A(G43S) fails to interact with its effector Melanophilin, indicating that the switch I region functions in the recruitment of Rab effector proteins. PMID- 18397839 TI - Mechanism of opioid-induced pupillary effects. AB - OBJECTIVE: This study tested the hypothesis that increased activity in the pupilloconstrictor nucleus by the addition of ambient light and by the administration of fentanyl, sufficient to block pupillary reflex dilation, constricts the pupil of anesthetized patients. METHODS: Pupil diameter was measured in 10 anesthetized patients during noxious stimulation above an epidural block level, in darkness and then with light directed into the left eye. Two measurements were taken from the right eye separated by 5 min. Following the second measurement, fentanyl (1 mcg/kg) was administered and the measures in light and dark were repeated. The effect of light and fentanyl on pupil size and pupillary reflex dilation were analyzed. RESULTS: An increase in light directed into the left eye constricted the pupil from 2.15+/-0.38 to 1.87+/-0.40 mm before fentanyl. Fentanyl did not constrict the pupil either in darkness or light but it did decrease pupillary reflex dilation by 49%. CONCLUSIONS: The miotic pupil during general anesthesia is not maximally constricted. Increased excitation of the pupilloconstrictor nucleus does not account for blockade of pupillary reflex dilation after fentanyl administration during desflurane anesthesia. SIGNIFICANCE: This study does not support the hypothesis that opioid effects on the human pupil are brought about by a direct excitatory action on the pupilloconstrictor nucleus. PMID- 18397840 TI - Changes in the activity of the cerebral cortex relate to postural response modification when warned of a perturbation. AB - OBJECTIVE: To determine whether the cerebral cortex contributes to modifying upcoming postural responses to external perturbations when provided with prior warning of the perturbation. METHODS: Electroencephalographic (EEG) potentials were recorded from 12 healthy human subjects (21-32 years of age) before perturbing their balance with backward translations of a platform under their feet. The subjects responded with and without a visual cue that warned them 2s before the perturbation (the Cue and No Cue conditions, respectively). RESULTS: Contingent negative variation (CNV) was evident before perturbation onset in only the Cue condition. In the Cue condition, the subjects also produced smaller center of pressure (CoP) displacements than in the No Cue condition. The cue related difference in the subjects' CNV potentials correlated with the cue related difference in their CoP displacements. No significant associations existed among the CNV potentials and any cue-related postural adjustments made before the perturbation. CONCLUSIONS: Cortical activity before an externally triggered perturbation associates with modifications of the ensuing postural response. SIGNIFICANCE: This is the first study to demonstrate a cortical correlate for changes in central postural set that modify externally triggered postural responses based on anticipation. PMID- 18397841 TI - Assessment of somatic complaints in environmental health. AB - In patients attributing their health complaints to environmental factors (EnvPat) evidence based medical diagnostics usually do not confirm environmental and somatic causes of symptoms. Many symptoms remain unexplained. Aim of the study was the systematic assessment of medically unexplained physical symptoms (MUPS) in EnvPat and comparison to symptom rates reported by subjects of an environmental study exposed to environmental odors (EnvExp). This specific exposure was chosen, as odors are associated by an unclear mechanism with physical symptoms. By this we aimed to enlighten the open question as to likeliness that MUPS of EnvPat are caused by hitherto unrevealed environmental exposures or result from somatization. MUPS were measured with SOMS-2 in EnvPat n=92, patients presenting in a university environmental outpatients clinic, and different study groups exposed to environmental odors (EnvExp). These were: (1) subjects exposed to annoying odors and medically relevant concentrations of bioaerosols, such as airborne microorganisms (EnvExp-1, n=74), and (2) subjects exposed to odors alone (EnvExp-2, n=282) as well as unexposed controls (Controls, n=235). Logistic regression and analysis of variance were applied to analyze rates of single complaints and the sum index of complaints (SOMS-CoIx). In EnvPat rates of MUPS were highest - significant (p<0.05) adjusted OR in 23 of 25 MUPS compared to controls - and highest SOMS-CoIx (mean 15.3 (S.D. +/-9.3). Rates of MUPS were lower in environmentally exposed subjects with difference in the two strata: while EnvExp-1 differed in several complaints, i.e., nausea and SOMS-CoIx (mean 7.2, S.D. +/-6.9) from controls (p<0.05), EnvExp-2 (SOMS-CoIx mean 4.8, S.D. +/-5.2) showed relevant differences only in two single complaints and not in the SOMS-CoIx from controls, SOMS-CoIx mean 3.9, S.D. +/-5.0. This remained when adjusting for age, gender, and school education. Rates of MUPS in environmental patients were clearly higher than in subjects with actual environmental exposure, making it unlikely that their symptoms are due to undetected environmental factors. MUPS of EnvPat show similarities to psychosomatic patients. In the environmental survey symptom assessment by SOMS-2 was sensitive to different environmental scenarios, i.e., higher rates of physical complaints were only found in subjects with hazardous residential bioaerosols pollution as well as an annoying odor exposure and interestingly not in subjects exposed to annoying odors alone. This underlines that questionnaire data of somatic complaints need to be interpreted on the basis of exposure assessment in order to unjustly attribute health complaints to annoyance. PMID- 18397842 TI - Cardiology clinical trial participation in community-based healthcare systems: obstacles and opportunities. AB - BACKGROUND: The objective of our study was to examine cardiologists' and organizational leaders' interest in clinical trial participation and perceived barriers and facilitators to participation within ten diverse non-profit healthcare delivery systems. Trials play a pivotal role in advancing knowledge about the safety and efficacy of cardiovascular interventions and tests. Although cardiovascular trials successfully enroll patients, recruitment challenges persist. Community-based health systems could be an important source of participants and investigators, but little is known about community cardiologists' experiences with trials. METHODS: We interviewed 25 cardiology and administrative leaders and mailed questionnaires to all 280 cardiologists at 10 U.S. healthcare organizations. RESULTS: The survey received a 73% response rate. While 60% of respondents had not participated in any trials in the past year, nearly 75% wanted greater participation. Cardiologists reported positive attitudes toward trial participation; more than half agreed that trials were their first choice of therapy for patients, if available. Almost all leaders described their organizations as valuing research but not necessarily trials. Major barriers to participation were lack of physician time and insufficient skilled research nurses. CONCLUSIONS: Cardiologists have considerable interest in trial participation. Major obstacles to increased participation are lack of time and effective infrastructure to support trials. These results suggest that community-based health systems are a rich source for cardiovascular research but additional funding and infrastructure are needed to leverage this resource. PMID- 18397843 TI - Recruitment to publicly funded trials--are surgical trials really different? AB - BACKGROUND: Good recruitment is integral to the conduct of a high-quality randomised controlled trial. It has been suggested that recruitment is particularly difficult for evaluations of surgical interventions, a field in which there is a dearth of evidence from randomised comparisons. While there is anecdotal speculation to support the inference that recruitment to surgical trials is more challenging than for medical trials we are unaware of any formal assessment of this. In this paper, we compare recruitment to surgical and medical trials using a cohort of publicly funded trials. DATA: Overall recruitment to trials was assessed using of a cohort of publicly funded trials (n=114). Comparisons were made by using the Recruitment Index, a simple measure of recruitment activity for multicentre randomised controlled trials. Recruitment at the centre level was also investigated through three example surgical trials. RESULTS: The Recruitment Index was found to be higher, though not statistically significantly, in the surgical group (n=18, median=38.0 IQR (10.7, 77.4)) versus (n=81, median=34.8 IQR (11.7, 98.0)) days per recruit for the medical group (median difference 1.7 (-19.2, 25.1); p=0.828). For the trials where the comparison was between a surgical and a medical intervention, the Recruitment Index was substantially higher (n=6, 68.3 (23.5, 294.8)) versus (n=93, 34.6 (11.7, 90.0); median difference 25.9 (-35.5, 221.8); p=0.291) for the other trials. CONCLUSIONS: There was no clear evidence that surgical trials differ from medical trials in terms of recruitment activity. There was, however, support for the inference that medical versus surgical trials are more difficult to recruit to. Formal exploration of the recruitment data through a modelling approach may go some way to tease out where important differences exist. PMID- 18397844 TI - Development and controversies of adjuvant therapy for pancreatic cancer. AB - BACKGROUND: Pancreatic cancer is an aggressive malignancy with a dismal prognosis. Radical surgery provides the only chance for a cure with a 5-year survival rate of 7%-25%. An effective adjuvant therapy is urgently needed to improve the surgical outcome. This review describes the current status of adjuvant therapy for pancreatic cancer, and highlights its controversies. DATA SOURCES: A Medline database search was performed to identify relevant articles using the keywords "pancreatic neoplasm", and "adjuvant therapy". Additional papers were identified by a manual search of the references from the key articles. RESULTS: Eight prospective randomized controlled trials (RCTs) on the use of adjuvant chemotherapy and chemoradiation for pancreatic cancer could be identified. The results for adjuvant regimens based on systemic 5-fluorouracil with or without external radiotherapy were conflicting. The recent two RCTs on gemcitabine based regimen gave promising results. CONCLUSIONS: Based on the available data, no standard adjuvant therapy for pancreatic cancer can be established yet. The best adjuvant regimen remains to be determined in large scale RCTs. Future trials should use a gemcitabine based regimen. PMID- 18397845 TI - Present status and recent advances in living donor liver transplantation for malignant hepatic tumors. AB - BACKGROUND: Living donor liver transplantation (LDLT) has been increasingly used to treat hepatic tumors worldwide in recent years, and is currently the most effective alternative to deceased donor liver transplantation to overcome the problem of organ shortage. LDLT has played an enormous role in treating early malignant hepatic tumors. But the indication of LDLT for malignant hepatic tumors is based on indefinite criteria. This review summarizes the recent studies in LDLT for treating malignant hepatic tumors. DATA SOURCES: A literature research of the PubMed database was conducted and research articles were reviewed. RESULTS: The current data on LDLT for malignant hepatic tumors, combined with our hospital experience, indicated that if a patient with hepatocellular carcinoma (HCC) who meets with the conventional Milan criteria cannot undergo tumor resection because of poorly preserved liver function, and a cadaveric graft is difficult to obtain within six months, LDLT may be selected. In a patient with recurrence of HCC after conventional therapies, feasibility, optimal timing, and efficacy of LDLT as a second-line treatment should be determined. CONCLUSIONS: Tumor recurrence is related to the biological behavior and staging of the tumor. New immunosuppressors which have anti-tumor effects and inhibit the immune system need to be developed. The indications of LDLT for hepatic malignant tumors should be selected meticulously. PMID- 18397846 TI - Use of health-related quality of life tools in hepatobiliary surgery. AB - BACKGROUND: The objective of this article is to review the literature and discuss the various tools used in hepatobiliary surgery for the measurement of health related quality of life (HR-QOL) and highlight various outcome variables that affect the HR-QOL among patients with common hepatobiliary disorders. DATA SOURCES: We reviewed HR-QOL articles published in the last 20 years on different hepatobiliary curative or palliative procedures in all languages. RESULTS: HR-QOL is a questionnaire tool which is utilized to assess the changes in the health status of patients after a hepatobiliary intervention. These surveys are of increasingly importance, as health care providers are challenged to justify treatment approaches and rationale for any surgical intervention. These HR-QOL tools are very helpful for the evaluation of subjective outcome of common hepatobiliary procedures like gastrointestinal quality of life index (GIQLI) for cholecystectomy, functional assessment in cancer therapy (FACT) for liver resection, short form 36 (SF-36) for liver transplantation, and quality of life questionnaire for patients with pancreatic cancer (QLQ-PAN). CONCLUSIONS: Use of validated and reliable health instruments in hepatobiliary surgery is directed at measuring the impact in a reproducible and valid fashion. Curative or palliative procedures should be offered to the patients of hepatobiliary disorders after the assessment by HR-QOL tools. Because the impairments of function that may occur after different operations vary considerably, an operation-specific assessment of HR-QOL for each type of surgical procedure is becoming an essential principle to follow in a successful healthcare system. PMID- 18397847 TI - Continuous versus bolus cardiac output monitoring during orthotopic liver transplantation. AB - BACKGROUND: Cardiac output monitoring is important for critical patients. This study aimed to determine the delayed response of continuous cardiac output (CCO) thermodilution measurement, whether CCO and bolus cardiac output (BCO) thermodilution agree sufficiently to be used interchangeably, and whether CCO monitoring is reliable for patients undergoing liver transplantation. METHODS: Thirteen patients undergoing liver transplantation without veno-venous bypass were studied (37-66 years old, weight 46-75 kg). Continuous and bolus thermodilution measurements were performed at predefined time points using an "Opti-Q" SvO2/CCO monitor (Abbott Laboratories, North Chicago, IL, USA). Bias and 95% limits of agreement were calculated according to Bland and Altman analysis. The limits of agreement by which two methods are judged to be interchangeable were defined in advance as +/-(13%XBCOmean) L/min. The repeatability and relative error of CCO, and the differences between CCO and the mean of the two measurements were calculated. RESULTS: Cardiac output measurements yielded 196 data pairs with ranges of 1.9 to 17.9 L/min for CCO and 2.1 to 18.3 L/min for BCO. The response time of CCO was delayed in the early phases after caval clamping and after reperfusion. At most of the measurement points, bias and 95% limits of agreement were -0.18+/-1.91 L/min. 95% limits of agreement did not fall within the predetermined limits of agreement of +/-1.14 L/min. The repeatability coefficient of CCO was 0.36 L/min and the relative error was 4.6+/-4.7%. The mean difference between CCO and the average of the two methods was -0.09 L/min (0.49 L/min). CONCLUSIONS: In patients undergoing liver transplantation, the delayed response of CCO limits its application during the early phases after caval clamping and after reperfusion of the graft. The two methods are not interchangeable even in hemodynamic stability. Continuous thermodilution monitoring, however, is reliable or acceptable for clinical purposes. PMID- 18397848 TI - Risk factors for fatal recurrence of hepatocellular carcinoma and their role in selecting candidates for liver transplantation. AB - BACKGROUND: With the maturation and popularization of skills in liver transplantation (LT), patients with hepatocellular carcinoma (HCC) have an alternative choice. LT as a curative treatment for HCC provides good liver function and systemic condition to recipients. Preoperative tumor characteristics are critical in selecting optimal candidates for LT to optimize the use of donor livers and to achieve a long-term survival. The present study aimed to elucidate the risk factors of HCC involved in fatal recurrence in the first year after LT and to investigate their utility in selecting suitable candidates for LT. METHODS: From April 2002 to October 2005, 303 patients who had received orthotopic LT for HCC were reviewed. Of these patients, those with diffuse intrahepatic or multiple systemic recurrent lesions who died within 1 year after surgery were investigated as fatal recurrence group (48 patients) and the remaining patients including those who were disease-free without recurrence, those who were alive with recurrence in the first year, or those who died in the first year of other causes, served as control group (255). The two groups were compared by demographics, tumor, and histopathological characteristics for their prognostic significance by logistic regression analysis. RESULTS: Multivariate analysis between the fatal recurrence group and the control group showed that the presence of vascular invasion, a tumor size greater than 6.5 cm, and a preoperative serum alpha-fetoprotein (AFP) level greater than 1000 mug/L were risk factors for fatal recurrence. Increased risk factors reduced the suitability of candidates for LT and diminished survival in the first year. 85.71% of the patients with all three risk factors, 37.84% of those with two factors, 13.64% of those with one factor, and 6.71% of those without risk factors died from tumor recurrence within 1 year after transplantation. CONCLUSIONS: Vascular invasion, tumor size > or =6.5 cm, and preoperative serum AFP level > or =1000 microg/L were significant predictors of fatal recurrence after LT for HCC. Patients with two or more risk factors should not be candidates for LT. PMID- 18397849 TI - Long-term outcome of gastric access loop in hepaticojejunostomy. AB - BACKGROUND: The incidence of hepaticojejunostomy stricture is 4%-10% in experienced centres. Many access loops have been designed and used to facilitate endoscopic intervention for this complication of hepaticojejunostomy. In this study, we investigated the effectiveness and safety of gastric access loop. METHODS: A retrospective analysis was conducted on 13 patients who had undergone hepaticojejunostomy with gastric access loop between June 1999 and September 2003. Eleven patients were followed up for a mean period of 51 months (range 20 81 months). Two patients were lost to follow up. RESULTS: On follow-up, 8 patients had patent jejunogastrostomy (end to side anastomosis between Roux loop of jejunum and stomach) and hepaticojejunostomy. Three patients developed stricture of jejunogastrostomy at 41 months, 63 months and 81 months of follow up. Among these 3 patients, one also had hepaticojejunostomy stricture. In the patient with hepaticojejunostomy stricture, dilatation of jejunogastrostomy stricture was attempted but failed. None of the patients had any evidence of bile gastritis/cholangitis. There was no procedure related morbidity/mortality. CONCLUSIONS: Further studies involving large numbers of patients are required before wide application of gastric access loop in hepaticojejunostomy though it is a safer option than percutaneous transhepatic manipulations or revision surgery. PMID- 18397850 TI - Use of surgical techniques in the rat pancreas transplantation model. AB - BACKGROUND: Pancreas transplantation is currently considered to be the most reliable and effective treatment for insulin-dependent diabetes mellitus (also called type 1 diabetes). With the improvement of microsurgical techniques, pancreas transplantation in rats has been the major model for physiological and immunological experimental studies in the past 20 years. We investigated the surgical techniques of pancreas transplantation in rats by analysing the difference between cervical segmental pancreas transplantation and abdominal pancreaticoduodenal transplantation. METHODS: Two hundred and forty male adult Wistar rats weighing 200-300 g were used, 120 as donors and 120 as recipients. Sixty cervical segmental pancreas transplants and 60 abdominal pancreaticoduodenal transplants were carried out and vessel anastomoses were made with microsurgical techniques. RESULTS: The time of donor pancreas harvesting in the cervical and abdominal groups was 31+/-6 and 37.6+/-3.8 min, respectively, and the lengths of recipient operations were 49.2+/-5.6 and 60.6+/-7.8 min. The time for donor operation was not significantly different (P>0.05), but the recipient operation time in the abdominal group was longer than that in the cervical group (P<0.05). In isograft transplantation without any preconditioning, 1-week survival rates were 93.6% and 86.2% in the cervical and abdominal groups, respectively, and were not significantly different (P>0.05). CONCLUSIONS: Both pancreas transplantation methods are stable models for immunological and physiological studies in pancreas transplantation. Since each has its own advantages and disadvantages, the designer can choose the appropriate method according to the requirements of the study. PMID- 18397851 TI - Etiology of fulminant hepatic failure: experience from a tertiary hospital in Bangladesh. AB - BACKGROUND: Fulminant hepatic failure (FHF) is not uncommon in our clinical practice in Bangladesh. There was a rise in acute hepatitis E virus (HEV) in Bangladesh after the 2004 floods. At that time, most of the country was under water for more than a month, leading to sewage contamination of the water supply. The aim of this study was to investigate the etiology of FHF in Bangladesh. METHODS: In this retrospective study, 23 patients with FHF who presented with severe impairment of hepatocellular function (i.e. encephalopathy, coagulopathy and jaundice) within 6 months of onset of symptoms were included. There were 17 men and 6 women, aged from 18 to 32 years. Four of the women were pregnant. Patients were tested for markers for common hepatotrophic viruses. A relevant history was taken and the Patient Record Book of the Unit was reviewed. RESULTS: 56.52% patients (13/23) had HEV infection, and all were anti-HEV IgM-positive tested by ELISA. HBV infection was detected in 34.78% patients (8/23), all of whom were tested positive for either HBsAg or anti-HBs IgM by ELISA. 8.7% patients (2/23) had a positive history for intake of alcohol and/or drugs. CONCLUSIONS: Acute HEV infection is the leading cause of FHF in Bangladesh. Sewage contamination of the water supply following floods contributes to a higher incidence of HEV infection. HBV infection is also important. PMID- 18397852 TI - Hepatic segmentectomy combined with major hepatic vein resection for preserving remnant liver lobe. AB - BACKGROUND: Impairment of liver function is the most serious complication that occurs after liver resection or in cirrhotic liver. Postoperative hepatic failure, which is mainly preceded by insufficient remnant liver function and/or postoperative septic complications, is the major cause of hospital mortality. This study was undertaken to evaluate hepatic segmentectomy combined with major hepatic vein (MHV) resection for preserving the remnant liver lobe in the treatment of resectable primary liver cancer. METHODS: From 1997 to 2007, six patients with primary liver cancer underwent hepatic segmentectomy with MHV resection, and three patients with hepatic vein injury had ligation of the MHV. The remnant liver lobe was preserved after hepatic segmentectomy combined with MHV resection or ligation. RESULTS: The preserved liver lobe with normal structure could maintain hepatic function and showed no evidence of atrophy or swelling after hepatic segmentectomy combined with MHV resection or ligation. CONCLUSIONS: After the right inferior hepatic vein is confirmed, and the MHV is occluded experimentally before hepatic segmentectomy combined with MHV resection, progressively deteriorating congestion does not occur in the preserved segment. Ligation or resection of the two MHVs must be avoided in patients with hepatic cirrhosis who have to undergo hepatic segmentectomy combined with MHV resection. Ligation of the MHV in patients with juxtahepatic vein injury is a simple and effective therapeutic modality. PMID- 18397853 TI - Ultrasonographic diagnosis of hepatic fungal infection. AB - BACKGROUND: Hepatic fungal infection is difficult to diagnose early because of the lack of related clinical knowledge. In this study, we analyzed the ultrasonic manifestations of hepatic fungal infection, in order to determine the diagnostic value of ultrasonography. METHODS: In 8 patients with hepatic fungal infection, the results of color Doppler flow images (CDFI) and contrast-enhanced ultrasonography (CEUS) were compared with those of ultrasound-guided biopsy. RESULTS: In the 8 patients with pathologically proven hepatic fungal infection, 5 showed multiple hypoechoic lesions with unequal sizes in the liver, commonly less than 2 cm in diameter. Some of the hypoechoic lesions showed a hyperechoic strip in the periphery, while a minority showed a "bull's eye" pattern, which is considered a typical manifestation of metastatic hepatic carcinoma. Solitary lesions in the right lobe of the liver were found in the other 3 patients. No blood flow signals were found in any lesions with CDFI. In the lesions a "filling defect" pattern was shown by CEUS, indicating no enhancement in the arterial phase, the portal vein phase and the delayed phase, except for 3 patients, in whom the lesions were slightly enhanced in the periphery during the early arterial phase. CONCLUSIONS: Hepatic fungal infection is characterized by an ultrasonic manifestation of multiple hypoechoic lesions with inequal size. A small number of lesions may show a characteristic "bull's eye" appearance. Moreover, a "filling defect" pattern on CEUS is helpful to make a correct diagnosis. In addition, ultrasound-guided biopsy is an important method for accurate diagnosis of hepatic fungal infection. PMID- 18397854 TI - Expression of transforming growth factors in hepatocellular carcinoma and its relations with clinicopathological parameters and prognosis. AB - BACKGROUND: Transforming growth factors (TGF)-beta1, TGF-betaR2 and Smad4 belong to the TGF family, and play important roles in carcinogenesis and the development of carcinoma, especially hepatocellular carcinoma (HCC). TGF-beta1 is a multipotent polypeptide, which inhibits the growth of epithelial cells including hepatoma cell lines and hepatocytes by inducing apoptosis. TGF-betaR2 forms a heterodimeric complex upon binding to TGF-beta, and then generates the first step in the signal transduction pathway leading to growth inhibition in coordination with the type 1 receptor. Smad4 protein is an important mediator in the TGF-beta signaling pathway, and negatively regulates the growth of epithelial cells. This study aimed to detect the expression of TGF-beta1, TGF-betaR2 and Smad4 in HCCs and their adjacent normal tissues, while assessing its relations with the clinicopathological parameters of HCC. METHODS: Forty-seven HCC specimens and their adjacent normal tissues were obtained surgically at the Affiliated Hospital of Medical College, Qingdao University. The expression of TGF-beta1, TGF-betaR2 and Smad4 was separately detected by immunohistochemistry in all HCC specimens and their adjacent normal tissues, and its relations with the clinicopathological parameters of HCC were assessed. RESULTS: The positive expression of TGF-beta1 was 72.34% in the HCC specimens, which was higher than that in the adjacent normal tissues (P<0.001). The positive expression of Smad4 and TGF-betaR2 was 34.04% and 59.57% respectively in the carcinoma specimens. The expression of TGF beta1, TGF-betaR2 and Smad4 was significantly higher in groups with a tumor embolus of the portal vein, integrity of the amicula, and Edmondson's III-IV than that in other groups, but it was not related to tumor size (P<0.05). CONCLUSIONS: TGF-beta1 may play an important role in the occurrence and development of HCC. Combined detection of TGF-beta1, TGF-betaR2 and Smad4 may be useful for the determination of the degree of malignancy and the prognosis of HCC. PMID- 18397855 TI - Effects of Dan-shao-hua-xian on expression of PPAR-gamma and NF-kappa B in rat liver fibrosis. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPAR-gamma) and nuclear factor kappa B (NF-kappaB) play important roles in liver fibrosis. This study aimed to investigate the effects of Dan-shao-hua-xian, a preparation of traditional Chinese medicine, on the expression of PPAR-gamma and NF-kappaB in the fibrotic livers of rats. METHODS: Seventy Wistar rats were randomly divided into 4 groups: treatment (model, 8 weeks+treatment, 8 weeks; group A), natural recovery ( model, 8 weeks+saline, 8 weeks; group B), model (model only, 8 weeks; group C), and control (normal, untreated, 16 weeks; group D). Each group consisted of 20 rats (except for group D, which had 10). Fibrotic liver models were induced in rats by subcutaneous injection of CCl4, oral administration of alcohol and a high-lipid/low-protein diet for 8 weeks. After the models were established, the rats in group A were orally given Dan-shao-hua-xian capsules daily for another 8 weeks. Then, the liver indices serum hyaluronic acid (HA), tumor necrosis factor-alpha (TNF-alpha) and alanine aminotransferase (ALT) were measured. The degree of hepatic fibrosis was evaluated by optical microscopy. Hydroxyproline (Hyp) in the liver tissue was determined. The expression of PPAR gamma was detected by immunohistochemical techniques. The protein levels of PPAR gamma and NF-kappaB were determined by Western blotting. RESULTS: The concentrations of serum HA, TNF-alpha and Hyp in group C increased compared with group D (P<0.05), and they decreased in group A compared with group C (P<0.05). The expression of PPAR-gamma in group C decreased compared with group D (P<0.05), and it increased in group A compared with groups B and C (P<0.05). Similarly, Western blotting showed that the expression of PPAR-gamma in group C decreased compared with group D, and it increased in group A compared with group C. The expression of NF-kappaB increased in group C compared with group D (P<0.05), and it decreased in group A compared with group C (P<0.05). CONCLUSION: Dan-shao-hua xian capsules enhance the expression of PPAR-gamma but decrease that of TNF-alpha and NF-kappaB in the liver tissues of CCl4-induced hepatic fibrotic rats. These effects may play a role in its activity in treating hepatic fibrosis. PMID- 18397856 TI - Activation of TLR-4 and liver injury via NF-kappa B in rat with acute cholangitis. AB - BACKGROUND: Toll-like receptors (TLRs) are a family of type 1 transmembrane receptors, which can recognize different pathogen-associated molecular patterns. Among them, TLR-4 is specific to lipopolysaccharide. It transfers the infection signal into the cell and promotes the translocation of nuclear factor kappa B (NF kappaB) to the nucleus and the subsequent transcriptional activation of genes encoding pro- and anti-inflammatory cytokines and chemokines. Acute cholangitis (AC) is a common biliary tract infection in oriental countries, and often leads to liver injury. The activation of TLR-4 and its significance in liver injury in rats with AC remain unclear. METHODS: Rat models of AC (biliary tract obstruction+E. coli injection, n=36) and control models (biliary tract obstruction+saline, n=18) were made. Liver tissue injury was investigated by pathological examination. The levels of serum TNF-alpha and IL-10 were measured by enzyme-linked immunosorbent assay, and the expressions of TLR-4, NF-kappaB mRNAs and proteins in the liver were detected by RT-PCR, immunohistochemical staining and Western blotting, respectively. RESULTS: Severe liver tissue injury in rats with AC was evident as shown by pathological examination. TLR-4 and NF kappaB were strongly expressed in the cytoplasm of hepatocytes in the AC group. They were negative or slightly positive in the control group. TLR-4 mRNA and protein in the liver of rats with AC increased 1 hour after biliary tract ligation and E. coli injection, and peaked at 6 hours after surgery. Twenty-four hours later, they began to decrease. The expression of TLR-4 was paralleled by that of NF-kappaB in the liver and TNF-alpha in serum. CONCLUSION: The higher expression of TLR-4 in the liver of rats with AC may be involved in liver injury through the activation of NF-kappaB and release of cytokines such as TNF-alpha. PMID- 18397857 TI - Exocrine pancreatic function assessed by secretin cholangio-Wirsung magnetic resonance imaging. AB - BACKGROUND: Magnetic resonance cholangiopancreatography (MRCP) is useful to assess exocrine pancreatic function by combining rapid imaging acquisition with the administration of secretin, a gastrointestinal hormone that stimulates the secretion of bile and pancreatic juice. However, extensive data on this method are lacking. This study aimed to determine whether MRCP with secretin administration is able to simultaneously detect alterations of both the pancreatic ducts and exocrine pancreatic function. METHODS: All subjects older than 18 years who underwent magnetic resonance imaging (MRI) and cholangio Wirsung magnetic resonance imaging (CWMRI) for suspicion of benign or malignant pancreatic diseases from January 2006 to December 2006 were enrolled in the study. MRI and CWMRI were carried out using a dedicated apparatus. RESULTS: Eighty-seven patients (46 males, 41 females, mean age 59.7+/-14.6, range 27-87 years) were enrolled. Of the 87 patients, 39 had a normal pancreas on imaging, 20 had an intrapapillary mucinous tumor (IPMT), and the rest had chronic pancreatitis (7), serous cystadenoma (6), a previous attack of acute biliary pancreatitis (5), congenital ductal abnormalities (5), mucinous cystadenoma (3), previous pancreatic head resection for autoimmune pancreatitis (1), or cholangiocarcinoma (1). Morphologically, we found two pseudocysts (one of the 7 patients with chronic pancreatitis, and one of the 5 patients after an attack of acute pancreatitis; the latter pseudocyst communicated with the main pancreatic duct). Calcifications were found in 3 of the 7 patients with chronic pancreatitis. All patients with IPMT and mucinous cystadenoma and 3 patients with serous cystadenoma were histologically confirmed. The remaining patients were followed up adequately to confirm the diagnosis by imaging. According to the Matos criteria, 73 patients (83.9%) were of grade 3, 8 grade 2, 4 grade 1, and 2 grade 0. The only pancreatic diseases which impaired the exocrine pancreatic secretion stimulated by secretin were chronic pancreatitis (57.1% of the patients, grade 0-1) and the IPMT mixed type in 2 of the 4 patients was grade 1. CONCLUSION: Secretin MRCP is a useful technique to simultaneously detect the presence of alterations of the pancreatic ducts and exocrine pancreatic function. PMID- 18397858 TI - Clinical analysis of solid-pseudopapillary tumor of the pancreas: report of 15 cases. AB - BACKGROUND: Solid-pseudopapillary tumor of the pancreas (SPTP) is an uncommon and enigmatic pancreatic neoplasm that occurs mainly in young women. Although more and more cases have been reported in recent years, misdiagnosis and incorrect treatment still frequently take place. This study was designed to stimulate consideration of this tumor. METHODS: We retrospectively reviewed the experience of diagnosis and treatment of 15 patients with SPTP and compared them with 516 patients with pancreatic cancer from January 1997 to March 2007. RESULTS: Most of the SPTP cases were asymptomatic except for one palpable mass. Almost all SPTPs demonstrated a solid structure with hypo- or iso-attenuation, cystic structure with hypo-attenuation on pre-contrast CT scan, and enhancement of solid portions on post-contrast CT scan. By contrast, most cases of pancreatic carcinoma had multiple symptoms and abnormal blood results. The tumors showed hypo-attenuation on both pre-contrast and post-contrast CT scan, and only a few showed iso attenuation on post-contrast CT scan. All cases of SPTP in our group were cured by surgical resection, while only 16.86% of patients with pancreatic carcinoma could undergo a radical resection. CONCLUSIONS: Clinical features and CT scans were helpful to differentiate SPTP from pancreatic carcinoma. Radical surgical resection was the most effective and safe method for the treatment of SPTP. PMID- 18397859 TI - The potential molecular mechanism of overexpression of uPA, IL-8, MMP-7 and MMP-9 induced by TRAIL in pancreatic cancer cell. AB - BACKGROUND: TNF-related apoptosis-inducing ligand (TRAIL) is a death ligand of the TNF-superfamily that has been implicated in inducing apoptosis in some tumor cells. The purpose of this study was to find out if TRAIL could induce the expression of uPA, IL-8, MMP-7 and MMP-9.and to explore the corresponding potential signaling transduction pathway in pancreatic cancer cells. METHODS: Colo357wt, Panc89 and PancTuI cell lines were stimulated by TRAIL (100 ng/ml). Crystal violet cell vitality assay was used to check the sensitivity to TRAIL induced apoptosis. Real-time RT-PCR tested the expression of uPA, IL-8, MMP-7 and MMP-9. RESULTS: TRAIL can stimulate the expression of uPA, IL-8, MMP-7 and MMP-9 in pancreatic cancer cell lines, especially in Colo357wt. The members of caspases, MEK1/2, PKC, and NF-kappaB are involved in TRAIL-induced expression of uPA, IL-8, MMP-7 and MMP-9. Furthermore, caspases play a different role in Colo357wt, Panc89 and PancTuI. CONCLUSIONS: TRAIL-treatment may result in the enhancement of invasion involving the signaling pathways of caspases, MEK1/2, PKC and NF-kappaB, in pancreatic cancer cells. It points to the necessity to carefully evaluate in vivo side effects of TRAIL. PMID- 18397860 TI - Malaria after living donor liver transplantation: report of two cases. AB - BACKGROUND: Infectious complications are common during the postoperative course of a liver transplant recipient. Malaria, however, is a rare complication in such a setting. METHOD: We report post-transplantation malaria causing elevation of liver enzymes in two recipients. RESULTS: Both patients who had undergone living donor liver transplantation showed elevated levels of liver enzymes and fever during the postoperative course. Investigations (including liver biopsy in one patient) were initially inconclusive in determining the cause of liver dysfunction. The diagnosis of malaria was established in both cases by peripheral blood smear. Liver function transiently worsened with antimalarial treatment but subsequently became normal. CONCLUSION: This report highlights the importance of excluding such uncommon causes of post-transplantation liver dysfunction, especially when either the recipient or the donor comes from a region endemic for malaria. PMID- 18397861 TI - Duodenum-preserving local excision of a gastrointestinal stromal tumor. AB - BACKGROUND: Duodenal gastrointestinal stromal tumors (GISTs) are rare. Because of the complex anatomy of the duodenum, the methods of resection of these tumors are controversial and diverse. METHODS: We report a case of a duodenal GIST in the anterolateral wall of the second part of the duodenum, which was successfully managed by local excision. The surgery was facilitated by preoperative mapping with Indian ink and navigation by endoscopy to assess the adequacy of resection and to avoid injury to the ampulla. RESULT: Reconstruction was successful with a duodenojejunostomy and protected by a nasoduodenal drain. CONCLUSION: The patient had no postoperative complications and the tumor was confirmed to be a GIST of the duodenum successful with an adequate resection margin. PMID- 18397862 TI - Vascular emergencies in cholelithiasis and cholecystectomy: our experience with two cases and literature review. AB - BACKGROUND: Complications from gallstones and laparoscopic cholecystectomy can be serious and fatal if there is a delay in recognition and treatment. We aim to present two unusual, life threatening vascular complications as a result of gallstones and laparoscopic cholecystectomy. Their management is highlighted with a brief review of literature. METHODS: Data for the article were gathered from clinical case note review. Radiology database was used for images. A brief literature review was undertaken using Pubmed search. The keywords used included hemobilia, pseudoaneurysm, arterio-biliary fistula and laparoscopic cholecystectomy. RESULTS: The article highlights two individual case reports. The first case constitutes an 81-year woman who had cystic arterial erosion causing hematemesis, while the second patient was a 57-year man who presented with hemobilia from a pseudoaneurysm of right hepatic artery (RHA) following laparoscopic cholecystectomy. Cystic arterial erosion was treated with subtotal cholecystectomy with duodenal defect closure while the pseudoaneurysm underwent radiological intervention. CONCLUSIONS: Cystic artery erosion and pseudoaneurysm causing arteriobiliary fistula are rare vascular complications related to the biliary tree. A high index of suspicion and timely intervention is important. Trauma to arteries should be avoided during laparoscopic cholecystectomy. PMID- 18397863 TI - Golden hamsters are nocturnal in captivity but diurnal in nature. AB - Daily activity rhythms are nearly universal among animals and their specific pattern is an adaptation of each species to its ecological niche. Owing to the extremely consistent nocturnal patterns of activity shown by golden hamsters (Mesocricetus auratus) in the laboratory, this species is a prime model for studying the mechanisms controlling circadian rhythms. In contrast to laboratory data, we discovered that female hamsters in the wild were almost exclusively diurnal. These results raise many questions about the ecological variables that shape the activity patterns in golden hamsters and the differences between laboratory and field results. PMID- 18397864 TI - Anterior-to-posterior wave of buccal expansion in suction feeding fishes is critical for optimizing fluid flow velocity profile. AB - In fishes that employ suction feeding, coordinating the timing of peak flow velocity with mouth opening is likely to be an important feature of prey capture success because this will allow the highest forces to be exerted on prey items when the jaws are fully extended and the flow field is at its largest. Although it has long been known that kinematics of buccal expansion in feeding fishes are characterized by an anterior-to-posterior wave of expansion, this pattern has not been incorporated in most previous computational models of suction feeding. As a consequence, these models have failed to correctly predict the timing of peak flow velocity, which according to the currently available empirical data should occur around the time of peak gape. In this study, we use a simple fluid dynamic model to demonstrate that the inclusion of an anterior-to-posterior wave of buccal expansion can correctly reproduce the empirically determined flow velocity profile, although only under very constrained conditions, whereas models that do not allow this wave of expansion inevitably predict peak velocity earlier in the strike, when the gape is less than half of its maximum. The conditions that are required to produce a realistic velocity profile are as follows: (i) a relatively long time lag between mouth opening and expansion of the more posterior parts of the mouth, (ii) a short anterior portion of the mouth relative to more posterior sections, and (iii) a pattern of movement that begins slowly and then rapidly accelerates. Greater maximum velocities were generated in simulations without the anterior-to-posterior wave of expansion, suggesting a trade-off between maximizing fluid speed and coordination of peak fluid speed with peak gape. PMID- 18397865 TI - Beyond robins: aerodynamic analyses of animal flight. AB - Recent progress in studies of animal flight mechanics is reviewed. A range of birds, and now bats, has been studied in wind tunnel facilities, revealing an array of wake patterns caused by the beating wings and also by the drag on the body. Nevertheless, the quantitative analysis of these complex wake structures shows a degree of similarity among all the different wake patterns and a close agreement with standard quasi-steady aerodynamic models and predictions. At the same time, new data on the flow over a bat wing in mid-downstroke show that, at least in this case, such simplifications cannot be useful in describing in detail either the wing properties or control prospects. The reasons for these apparently divergent results are discussed and prospects for future advances are considered. PMID- 18397866 TI - Global patterns in the phylogenetic structure of island mammal assemblages. AB - Assemblage-level phylogenies carry the signature of ecological and evolutionary processes, which may provide useful information on modes of assemblage formation. We present a global-scale analysis of the emergent phylogenetic properties of mammal assemblages on islands, in which we compared the structure of 595 island assemblages with null models constructed under four alternative definitions of regional source pools. Although most assemblages had a structure indistinguishable from random samples, for some mammal taxa, up to 40% of island assemblages were phylogenetically overdispersed. This suggests that in at least some cases, the processes that shape island faunas are not independent of phylogeny. Furthermore, measures of phylogenetic structure were associated in some cases with island geographical features (size, maximum elevation and habitat diversity). Our results suggest that part of the signal of assemblage formation processes is detectable in the phylogenies of contemporary island mammal faunas, though much is obscured by the complexity of these processes. PMID- 18397867 TI - Macrophysiology for a changing world. AB - The Millennium Ecosystem Assessment (MA) has identified climate change, habitat destruction, invasive species, overexploitation and pollution as the major drivers of biodiversity loss and sources of concern for human well-being. Understanding how these drivers operate and interact and how they might be mitigated are among the most pressing questions facing humanity. Here, we show how macrophysiology--the investigation of variation in physiological traits over large geographical, temporal and phylogenetic scales--can contribute significantly to answering these questions. We do so by demonstrating, for each of the MA drivers, how a macrophysiological approach can or has helped elucidate the impacts of these drivers and their interactions. Moreover, we illustrate that a large-scale physiological perspective can provide insights into previously unrecognized threats to diversity, such as the erosion of physiological variation and stress tolerance, which are a consequence of the removal of large species and individuals from the biosphere. In so doing we demonstrate that environmental physiologists have much to offer the scientific quest to resolve major environmental problems. PMID- 18397868 TI - Breeding synchrony in colonial birds: from local stress to global harmony. AB - Why and how birds in colonies often breed in striking synchrony is an unsolved question. In colonies, conspecific birds often destroy eggs and kill chicks, either intentionally or not. We propose that social tranquillity at the time of laying can be achieved if a bird's stress level is partly determined by the agitation of its neighbours. Moreover, we propose that this local process, together with environmental cues, can synchronize breeding between neighbours and through a whole colony. We tested our hypotheses using a generic individual-based model where the breeding predisposition of females was updated daily depending on an increase in the photoperiod (positively) and the stress level of neighbours: negatively if they were agitated, and positively otherwise. A female laid her eggs when her stress level fell to a critical value. Even giving only a low relevance to the neighbour's stress level was enough to synchronize the laying date of neighbours and also of a huge colony. Moreover, females bred in a safer environment, which is known from field studies to increase fitness. Our study highlights the power of local adaptive (individual) behaviour to create global (colony) patterns. We argue that collective patterns such as breeding synchrony in colonial birds could have simple adaptive individual-level explanations. PMID- 18397869 TI - Mapping the spawning grounds of North Sea cod (Gadus morhua) by direct and indirect means. AB - Despite recent evidence for sub-stock structuring, North Sea cod are assessed as a single unit. As a consequence, knowledge of sub-stock trends is poor. In particular, there are no recent evaluations of which spawning grounds are active. Here we report results from the first ichthyoplankton survey to cover the whole North Sea. Also, this survey, conducted in 2004, was the first to make extensive use of DNA-based molecular methods to unambiguously identify early developmental stage cod eggs. We compare the findings from the plankton survey with estimated egg production inferred from the distribution of mature cod in contemporaneous trawl surveys. Results from both approaches were in general agreement and showed hot spots of egg production around the southern and eastern edges of the Dogger Bank, in the German Bight, the Moray Firth and to the east of the Shetlands. These areas broadly coincide with known spawning locations from the period 1940 to 1970. We were, however, unable to directly detect significant numbers of cod eggs at the historic spawning ground off Flamborough (northeast coast of England). The results demonstrate that most of the major spawning grounds of cod in the North Sea are still active but that some localized populations may have been reduced to the point where it is now difficult to detect the presence of eggs in the plankton. PMID- 18397870 TI - Early nutrition and phenotypic development: 'catch-up' growth leads to elevated metabolic rate in adulthood. AB - Resting metabolic rate (RMR) is responsible for up to 50% of total energy expenditure, and so should be under strong selection pressure, yet it shows extensive intraspecific variation and a low heritability. Environmental conditions during growth are thought to have long-term effects through 'metabolic programming'. Here we investigate whether nutritional conditions early in life can alter RMR in adulthood, and whether this is due to growth acceleration or the change in diet quality that prompts it. We manipulated dietary protein levels during the main growth period of zebra finches (Taeniopygia guttata) such that an episode of poor nutrition occurred with and without growth acceleration. This produced different growth trajectories but a similar adult body mass. Only the diet that induced growth acceleration resulted in a significant (19%) elevation of RMR at adulthood, despite all the birds having been on the same diet after the first month. This is the first study to show that dietary-induced differences in growth trajectories can have a long-term effect on adult metabolic rate. It suggests that modification of metabolic efficiency may be one of the mechanisms mediating the observed long-term costs of accelerated growth, and indicates links between early nutrition and the metabolic syndrome. PMID- 18397871 TI - Feasibility of combined use of intravascular ultrasound radiofrequency data analysis and optical coherence tomography for detecting thin-cap fibroatheroma. AB - AIMS: To evaluate the feasibility of the combined use of virtual histology (VH) intravascular ultrasound (IVUS) and optical coherence tomography (OCT) for detecting in vivo thin-cap fibroatheroma (TCFA). METHODS AND RESULTS: In 56 patients with angina, 126 plaques identified by IVUS findings were analysed using both VH-IVUS and OCT. IVUS-derived TCFA was defined as an abundant necrotic core (>10% of the cross-sectional area) in contact with the lumen (NCCL) and %plaque volume >40%. OCT-derived TCFA was defined as a fibrous cap thickness of <65 microm overlying a low-intensity area with an unclear border. Plaque meeting both TCFA criteria was defined as definite-TCFA. Sixty-one plaques were diagnosed as IVUS-derived TCFA and 36 plaques as OCT-derived TCFA. Twenty-eight plaques were diagnosed as definite-TCFA; the remaining 33 IVUS-derived TCFA had a non-thin-cap and eight OCT-derived TCFA had a non-NCCL (in discord with NCCL visualized by VH IVUS, mainly due to misreading caused by dense calcium). Based on IVUS findings, definite-TCFA showed a larger plaque and vessel volume, %plaque-volume, higher vessel remodelling index, and greater angle occupied by the NCCL in the lumen circumference than non-thin-cap IVUS-derived TCFA. Conclusion Neither modality alone is sufficient for detecting TCFA. The combined use of OCT and VH-IVUS might be a feasible approach for evaluating TCFA. PMID- 18397872 TI - Downregulation of the calcium current in human right atrial myocytes from patients in sinus rhythm but with a high risk of atrial fibrillation. AB - AIMS: A decrease in L-type calcium current (ICaL) is an important mechanism favouring atrial fibrillation (AF). Here, we aimed to identify pathogenic factors associated with ICaL downregulation. METHODS AND RESULTS: Atrial myocytes were isolated from right atrial appendages obtained from 86 adult patients in sinus rhythm with coronary artery disease, aortic valve disease, or mitral valve disease (MVD). Current was recorded in isolated myocytes using the whole-cell patch-clamp technique. The ICaL recorded in the 172 myocytes studied showed a marked variability of peak density ranging from 0.1 to 9.0 pA/pF. The ICaL peak density did not correlate with membrane capacitance or changes in current biophysical properties. The ICaL peak density was homogeneous for a given sample. Small ICaL values were recorded in patients with MVD or with a low left ventricular ejection fraction (<45%). Small ICaL values were more sensitive to the beta-adrenergic agonist, isoproterenol (1 microM), and to the phosphodiesterase inhibitor, 3-isobutyl-1-methyl-xanthine (10 microM). CONCLUSION: In human atrial myocytes, the variability of ICaL is related to the clinical history of the donors. The downregulation of ICaL is already observed in patients in sinus rhythm with a high risk of AF and is associated with the greatest response to beta-adrenergic agonist. PMID- 18397873 TI - Evaluating the impact of atrial dilatation on atrial calcium cycling. PMID- 18397874 TI - Prognosis, disease progression, and treatment of atrial fibrillation patients during 1 year: follow-up of the Euro Heart Survey on atrial fibrillation. AB - AIMS: To gain insight in the prognosis and treatment of atrial fibrillation (AF) patients during 1-year follow-up in the Euro Heart Survey (EHS) on AF. METHODS AND RESULTS: The EHS enrolled 5333 AF patients in 2003--2004. One-year follow-up data were available for 80%. Of first detected AF patients, 46% did not have a recurrence during 1 year, paroxysmal AF largely remained paroxysmal AF (80%), and 30% of persistent AF progressed to permanent AF. Many treatment changes occurred since baseline. Oral anticoagulation was started in 19% and discontinued in 16% of all patients. Of patients initially on rhythm control 27% did not receive rhythm control during follow-up, whereas 15% of patients initially on rate control received rhythm control. Mortality was highest in permanent AF (8.2%), but also substantial in first detected AF (5.7%). In multivariable analysis, sinus rhythm at baseline was associated with lower mortality, but no significant effect was observed regarding the application of either rhythm or rate control. CONCLUSION: The EHS on AF provides unique prospective observational data on AF progression, long-term treatment, prognosis, and determinants of adverse outcome of the total clinical spectrum of AF in a European cardiology-based patient cohort. PMID- 18397875 TI - A sensitized mutagenesis screen identifies Gli3 as a modifier of Sox10 neurocristopathy. AB - Haploinsufficiency for the transcription factor SOX10 is associated with the pigmentary deficiencies of Waardenburg syndrome (WS) and is modeled in Sox10 haploinsufficient mice (Sox10(LacZ/+)). As genetic background affects WS severity in both humans and mice, we established an N-ethyl-N-nitrosourea (ENU) mutagenesis screen to identify modifiers that increase the phenotypic severity of Sox10(LacZ/+) mice. Analysis of 230 pedigrees identified three modifiers, named modifier of Sox10 neurocristopathies (Mos1, Mos2 and Mos3). Linkage analysis confirmed their locations on mouse chromosomes 13, 4 and 3, respectively, within regions distinct from previously identified WS loci. Positional candidate analysis of Mos1 identified a truncation mutation in a hedgehog(HH)-signaling mediator, GLI-Kruppel family member 3 (Gli3). Complementation tests using a second allele of Gli3 (Gli3(Xt-J)) confirmed that a null mutation of Gli3 causes the increased hypopigmentation in Sox10(LacZ/+);Gli3(Mos1/)(+) double heterozygotes. Early melanoblast markers (Mitf, Sox10, Dct, and Si) are reduced in Gli3(Mos1/)(Mos1) embryos, indicating that loss of GLI3 signaling disrupts melanoblast specification. In contrast, mice expressing only the GLI3 repressor have normal melanoblast specification, indicating that the full-length GLI3 activator is not required for specification of neural crest to the melanocyte lineage. This study demonstrates the feasibility of sensitized screens to identify disease modifier loci and implicates GLI3 and other HH signaling components as modifiers of human neurocristopathies. PMID- 18397876 TI - Sirtuin inhibition protects from the polyalanine muscular dystrophy protein PABPN1. AB - Oculopharyngeal muscular dystrophy (OPMD) is caused by polyalanine expansion in nuclear protein PABPN1 [poly(A) binding protein nuclear 1] and characterized by muscle degeneration. Druggable modifiers of proteotoxicity in degenerative diseases, notably the longevity modulators sirtuins, may constitute useful therapeutic targets. However, the modifiers of mutant PABPN1 are unknown. Here, we report that longevity and cell metabolism modifiers modulate mutant PABPN1 toxicity in the muscle cell. Using PABPN1 nematodes that show muscle cell degeneration and abnormal motility, we found that increased dosage of the sirtuin and deacetylase sir-2.1/SIRT1 exacerbated muscle pathology, an effect dependent on the transcription factor daf-16/FoxO and fuel sensor aak-2/AMPK (AMP-activated protein kinase), while null mutants of sir-2.1, daf-16 and aak-2 were protective. Consistently, the Sir2 inhibitor sirtinol was protective, whereas the Sir2 and AMPK activator resveratrol was detrimental. Furthermore, rescue by sirtinol was dependent on daf-16 and not aak-2, whereas aggravation by resveratrol was dependent on aak-2 and not daf-16. Finally, the survival of mammalian cells expressing mutant PABPN1 was promoted by sirtinol and decreased by resveratrol. Altogether, our data identify Sir2 and AMPK inhibition as therapeutic strategies for muscle protection in OPMD, extending the value of druggable proteins in cell maintenance networks to polyalanine diseases. PMID- 18397878 TI - Ablation of a focal left atrial tachycardia via a retrograde approach using remote magnetic navigation. PMID- 18397877 TI - Bladder tumour-derived somatic TSC1 missense mutations cause loss of function via distinct mechanisms. AB - More than 50% of transitional cell carcinomas of the bladder show loss of heterozygosity of a region spanning the TSC1 locus at 9q34 and mutations of TSC1 have been identified in 14.5% of tumours. These comprise nonsense mutations, splicing mutations, small deletions and missense mutations. Missense mutations are only rarely found in the germline in TSC disease. Therefore, we have examined six somatic missense mutations found in bladder cancer to determine whether these result in loss of function. We describe loss of function via distinct mechanisms. Five mutations caused mutually exclusive defects at mRNA and protein levels. Of these, two mutations caused pre-mRNA splicing errors that were predicted to result in premature protein truncation and three resulted in markedly reduced stability of exogenous TSC1 protein. Primary tumours with aberrant TSC1 pre-mRNA splicing were confirmed as negative for TSC1 expression by immunohistochemistry. Expression was also significantly reduced in a tumour with a TSC1 missense mutation resulting in diminished protein half-life. A single TSC1 missense mutation identified in a tumour with retained heterozygosity of the TSC1 region on chromosome 9 caused an apparently TSC2- and mTOR-independent localization defect of the mutant protein. We conclude that although TSC1 missense mutations do not play a major role in causation of TSC disease, they represent a significant proportion of somatic loss of function mutations in bladder cancer. PMID- 18397879 TI - Afi1p functions as an Arf3p polarization-specific docking factor for development of polarity. AB - ADP-ribosylation factors (Arfs) are highly conserved small GTPases and are critical components of vesicle trafficking. Yeast Arf3p, despite its similarity to mammalian Arf6, is not required for endocytosis but is involved in polarity development. In this study, we identified an Arf3p interacting protein 1 (Afi1p), which, through its N-terminal conserved region, specifically interacts with GTP bound Arf3p. Afi1p is distributed asymmetrically at the plasma membrane and is required for polarized distribution of Arf3p but not of an Arf3p guanine nucleotide-exchange factor, Yel1p. However, Afi1p is not required for targeting of Arf3p or Yel1p to the plasma membrane. Like arf3 mutant yeast, afi1 mutant yeast exhibited an abnormal budding pattern and partially delayed actin patch polarization. An Afi1p, (38)KLGP4A-Afi1p, mutated at the Arf3p-binding region, loses its ability to interact with Arf3p and maintain the polarized distribution of Arf3p. Although (38)KLGP4A-Afi1p still possessed a proper polarized distribution, it lost its ability to rescue actin patch polarization in afi1 mutant cells. Our findings demonstrate that Afi1p functions as an Arf3p polarization-specific adapter and participates in development of polarity. PMID- 18397880 TI - Activation of p300 histone acetyltransferase activity is an early endothelial response to laminar shear stress and is essential for stimulation of endothelial nitric-oxide synthase mRNA transcription. AB - Previous studies have shown that the acute stimulation of endothelial nitric oxide synthase (eNOS) mRNA transcription by laminar shear stress is dependent on nuclear factor kappa B (NFkappaB) subunits p50 and p65 binding to a shear stress response element (SSRE) in the human eNOS promoter and that mutation of the SSRE abrogates the shear-stimulated increase in eNOS promoter activity. In the present study, we found that although shear markedly increased eNOS mRNA, the increase in nuclear translocation of p50 and p65 caused by shear was only 2-fold, suggesting that shear has additional effects on NFkappaB cofactor activity beyond nuclear translocation. Chromatin immunoprecipitation assays showed that virtually no p50 or p65 was bound to the eNOS promoter at base line but that shear increased the binding of these subunits to the human eNOS SSRE by 10- to 20-fold. Co immunoprecipitation studies demonstrated during the first 30 min of shear p300 bound to p65. Shear also increased p300 histone acetyltransferase (HAT) activity by 2.5-fold and increased acetylation of p65. The increase in eNOS mRNA caused by shear was completely blocked by pharmacological inhibition of p300/HAT activity with curcumin or by p300 small interfering RNA. Chromatin immunoprecipitation assays also showed that shear stimulated acetylation of histones 3 and 4 at the region of the eNOS promoter SSRE and extended 3' toward the eNOS coding region. This was associated with opening of chromatin at the SSRE. In conclusion, these studies reveal a previously unknown role of p300/HAT activation as a very early response to shear that is essential for increasing eNOS mRNA levels. PMID- 18397882 TI - Suppression of inhibin A biological activity by alterations in the binding site for betaglycan. AB - Inhibins A and B negatively regulate the production and secretion of follicle stimulating hormone from the anterior pituitary, control ovarian follicle development and steroidogenesis, and act as tumor suppressors in the gonads. Inhibins regulate these reproductive events by forming high affinity complexes with betaglycan and activin or bone morphogenetic protein type II receptors. In this study, the binding site of inhibin A for betaglycan was characterized using inhibin A mutant proteins. An epitope for high affinity betaglycan binding was detected spanning the outer convex surface of the inhibin alpha-subunit. Homology modeling indicates that key alpha-subunit residues (Tyr(50), Val(108), Thr(111), Ser(112), Phe(118), Lys(119), and Tyr(120)) form a contiguous epitope in this region of the molecule. Disruption of betaglycan binding by the simultaneous substitution of Thr(111), Ser(112), and Tyr(120) to alanine yielded an inhibin A variant that was unable to suppress activin-induced follicle-stimulating hormone release by rat pituitary cells in culture. Together these results indicate that a high affinity interaction between betaglycan and residues Val(108)-Tyr(120) of the inhibin alpha-subunit mediate inhibin A biological activity. PMID- 18397881 TI - Two heparanase splicing variants with distinct properties are necessary in early Xenopus development. AB - Heparanase is an endoglycosidase that cleaves heparan sulfate (HS) side chains from heparan sulfate proteoglycans (HSPGs) present in extracellular matrix and cell membranes. Although HSPGs have many functions during development, little is known of the role of the enzyme that degrades HS, heparanase. We cloned and characterized the expression of two heparanase splicing variants from Xenopus laevis and studied their function in early embryonic development. The heparanase gene (termed xHpa) spans over 15 kb and consists of at least 12 exons. The long heparanase (XHpaL) cDNA encodes a 531-amino acid protein, whereas the short splicing variant (XHpaS) results in a protein with the same open reading frame but missing 58 amino acids as a consequence of a skipped exon 4. Comparative studies of both isoforms using heterologous expression systems showed: 1) XHpaL is enzymatically active, whereas XHpaS is not; 2) XHpaL and XHpaS interact with heparin and HS; 3) both proteins traffic through the endoplasmic reticulum and Golgi apparatus, but XHpaL is secreted into the medium, whereas XHpaS remains associated with the membrane as a consequence of the loss of three glycosylation sites; 4) overexpression of XHpaS but not XHpaL increases cell adhesion of glioma cells to HS-coated surfaces; 5) XHpaL and XHpaS mRNA and protein levels vary as development progresses; 6) specific antisense knock-down of both XHpaL and XHpaS, but not XHpaL alone, results in failure of embryogenesis to proceed. Interestingly, rescue experiments suggest that the two heparanases regulate the same developmental processes, but via different mechanisms. PMID- 18397883 TI - Transglutaminase induces protofibril-like amyloid beta-protein assemblies that are protease-resistant and inhibit long-term potentiation. AB - An increasing body of evidence suggests that soluble assemblies of amyloid beta protein (Abeta) play an important role in the initiation of Alzheimer disease (AD). In vitro studies have found that synthetic Abeta can form soluble aggregates through self-assembly, but this process requires Abeta concentrations 100- to 1000-fold greater than physiological levels. Tissue transglutaminase (TGase) has been implicated in neurodegeneration and can cross-link Abeta. Here we show that TGase induces rapid aggregation of Abeta within 0.5-30 min, which was not observed with chemical cross-linkers. Both Abeta40 and Abeta42 are good substrates for TGase but show different aggregation patterns. Guinea pig and human TGase induced similar Abeta aggregation patterns, and oligomerization was observed with Abeta40 concentrations as low as 50 nm. The formed Abeta40 species range from 5 to 6 nm spheres to curvilinear structures of the same width, but up to 100 nm in length, that resemble the previously described self-assembled Abeta protofibrils. TGase-induced Abeta40 assemblies are resistant to a 1-h incubation with either neprilysin or insulin degrading enzyme, whereas the monomer is rapidly degraded by both proteases. In support of these species being pathological, TGase-induced Abeta40 assemblies (100 nm) inhibited long term potentiation recorded in the CA1 region of mouse hippocampus slices. Our data suggest that TGase can contribute to AD by initiating Abeta oligomerization and aggregation at physiological levels, by reducing the clearance of Abeta due to the generation of protease-resistant Abeta species, and by forming Abeta assemblies that inhibit processes involved in memory and learning. Our data suggest that TGase might constitute a specific therapeutic target for slowing or blocking the progression of AD. PMID- 18397884 TI - Regulation of TIP60 by ATF2 modulates ATM activation. AB - TIP60 (HTATIP) is a histone acetyltransferase (HAT) whose function is critical in regulating ataxia-telangiectasia mutated (ATM) activation, gene expression, and chromatin acetylation in DNA repair. Here we show that under non-stressed conditions, activating transcription factor-2 (ATF2) in cooperation with Cul3 ubiquitin ligase promotes degradation of TIP60, thereby attenuating its HAT activity. Inhibiting either ATF2 or Cul3 expression by small interfering RNA stabilizes the TIP60 protein. ATF2 association with TIP60 on chromatin is decreased following exposure to ionizing radiation (IR), resulting in enhanced TIP60 stability and activity. We also identified a panel of melanoma and prostate cancer cell lines whose ATF2 expression is inversely correlated with TIP60 levels and ATM activation after IR. Inhibition of ATF2 expression in these lines restored TIP60 protein levels and both basal and IR-induced levels of ATM activity. Our study provides novel insight into regulation of ATM activation by ATF2-dependent control of TIP60 stability and activity. PMID- 18397885 TI - The GP(Y/F) domain of TF1 integrase multimerizes when present in a fragment, and substitutions in this domain reduce enzymatic activity of the full-length protein. AB - Integrases (INs) of retroviruses and long terminal repeat retrotransposons possess a C-terminal domain with DNA binding activity. Other than this binding activity, little is known about how the C-terminal domain contributes to integration. A stretch of conserved amino acids called the GP(Y/F) domain has been identified within the C-terminal IN domains of two distantly related families, the gamma-retroviruses and the metavirus retrotransposons. To enhance understanding of the C-terminal domain, we examined the function of the GP(Y/F) domain in the IN of Tf1, a long terminal repeat retrotransposon of Schizosaccharomyces pombe. The activities of recombinant IN were measured with an assay that modeled the reverse of integration called disintegration. Although deletion of the entire C-terminal domain disrupted disintegration activity, an alanine substitution (P365A) in a conserved amino acid of the GP(Y/F) domain did not significantly reduce disintegration. When assayed for the ability to join two molecules of DNA in a reaction that modeled forward integration, the P365A substitution disrupted activity. UV cross-linking experiments detected DNA binding activity in the C-terminal domain and found that this activity was not reduced by substitutions in two conserved amino acids of the GP(Y/F) domain, G364A and P365A. Gel filtration and cross-linking of a 71-amino acid fragment containing the GP(Y/F) domain revealed a surprising ability to form dimers, trimers, and tetramers that was disrupted by the G364A and P365A substitutions. These results suggest that the GP(Y/F) residues may play roles in promoting multimerization and intermolecular strand joining. PMID- 18397886 TI - Phosphorylation-dependent binding of 14-3-3 proteins controls TRESK regulation. AB - The two-pore domain K(+) channel, TRESK (TWIK-related spinal cord K(+) channel) is reversibly activated by the calcium/calmodulin-dependent protein phosphatase, calcineurin. In the present study, we report that 14-3-3 proteins directly bind to the intracellular loop of TRESK and control the kinetics of the calcium dependent regulation of the channel. Coexpression of 14-3-3eta with TRESK blocked, whereas the coexpression of a dominant negative form of 14-3-3eta accelerated the return of the K(+) current to the resting state after the activation mediated by calcineurin in Xenopus oocytes. The direct action of 14-3 3 was spatially restricted to TRESK, since 14-3-3eta was also effective, when it was tethered to the channel by a flexible polyglutamine-containing chain. The effect of both the coexpressed and chained 14-3-3 was alleviated by the microinjection of Ser(P)-Raf259 phosphopeptide that competes with TRESK for binding to 14-3-3. The gamma and eta isoforms of 14-3-3 controlled TRESK regulation, whereas the beta, zeta, epsilon, sigma, and tau isoforms failed to influence the mechanism significantly. Phosphorylation of serine 264 in mouse TRESK was required for the binding of 14-3-3eta. Because 14-3-3 proteins are ubiquitous, they are expected to control the duration of calcineurin-mediated TRESK activation in all the cell types that express the channel, depending on the phosphorylation state of serine 264. This kind of direct control of channel regulation by 14-3-3 is unique within the two-pore domain K(+) channel family. PMID- 18397887 TI - Protein phosphatase 2A is targeted to cell division control protein 6 by a calcium-binding regulatory subunit. AB - The cell division control protein 6 (Cdc6) is essential for formation of pre replication complexes at origins of DNA replication. Phosphorylation of Cdc6 by cyclin-dependent kinases inhibits ubiquitination of Cdc6 by APC/C(cdh1) and degradation by the proteasome. Experiments described here show that the PR70 member of the PPP2R3 family of regulatory subunits targets protein phosphatase 2A (PP2A) to Cdc6. Interaction with Cdc6 is mediated by residues within the C terminus of PR70, whereas interaction with PP2A requires N-terminal sequences conserved within the PPP2R3 family. Two functional EF-hand calcium-binding motifs mediate a calcium-enhanced interaction of PR70 with PP2A. Calcium has no effect on the interaction of PR70 with Cdc6 but enhances the association of PP2A with Cdc6 through its effects on PR70. Knockdown of PR70 by RNA interference results in an accumulation of endogenous and expressed Cdc6 protein that is dependent on the cyclin-dependent protein kinase phosphorylation sites on Cdc6. Knockdown of PR70 also causes G(1) arrest, suggesting that PR70 function is critical for progression into S phase. These observations indicate that PP2A can be targeted in a calcium-regulated manner to Cdc6 via the PR70 subunit, where it plays a role in regulating protein phosphorylation and stability. PMID- 18397888 TI - The Parkinson disease-associated leucine-rich repeat kinase 2 (LRRK2) is a dimer that undergoes intramolecular autophosphorylation. AB - Mutations in leucine-rich repeat kinase 2 (LRRK2) are a common cause of familial and apparently sporadic Parkinson disease. LRRK2 is a multidomain protein kinase with autophosphorylation activity. It has previously been shown that the kinase activity of LRRK2 is required for neuronal toxicity, suggesting that understanding the mechanism of kinase activation and regulation may be important for the development of specific kinase inhibitors for Parkinson disease treatment. Here, we show that LRRK2 predominantly exists as a dimer under native conditions, a state that appears to be stabilized by multiple domain-domain interactions. Furthermore, an intact C terminus, but not N terminus, is required for autophosphorylation activity. We identify two residues in the activation loop that contribute to the regulation of LRRK2 autophosphorylation. Finally, we demonstrate that LRRK2 undergoes intramolecular autophosphorylation. Together, these results provide insight into the mechanism and regulation of LRRK2 kinase activity. PMID- 18397890 TI - Molecular modeling of benzothiazepine binding in the L-type calcium channel. AB - Benz(othi)azepine (BTZ) derivatives constitute one of three major classes of L type Ca(2+) channel ligands. Despite intensive experimental studies, no three dimensional model of BTZ binding is available. Here we have built KvAP- and KcsA based models of the Ca(v)1.2 pore domain in the open and closed states and used multiple Monte Carlo minimizations to dock representative ligands. In our open channel model, key functional groups of BTZs interact with BTZ-sensing residues, which were identified in previous mutational experiments. The bulky tricyclic moiety occupies interface between domains III and IV, while the ammonium group protrudes into the inner pore, where it is stabilized by nucleophilic C-ends of the pore helices. In the closed channel model, contacts with several ligand sensing residues in the inner helices are lost, which weakens ligand-channel interactions. An important feature of the ligand-binding mode in both open and closed channels is an interaction between the BTZ carbonyl group and a Ca(2+) ion chelated by the selectivity filter glutamates in domains III and IV. In the absence of Ca(2+), the tricyclic BTZ moiety remains in the domain interface, while the ammonium group directly interacts with a glutamate residue in the selectivity filter. Our model suggests that the Ca(2+) potentiation involves a direct electrostatic interaction between aCa(2+) ion and the ligand rather than an allosteric mechanism. Energy profiles indicate that BTZs can reach the binding site from the domain interface, whereas access through the open activation gate is unlikely, because reorientation of the bulky molecule in the pore is hindered. PMID- 18397889 TI - Tumor suppressor protein p53 regulates megakaryocytic polyploidization and apoptosis. AB - The molecular mechanisms underlying differentiation of hematopoietic stem cells into megakaryocytes are poorly understood. Tumor suppressor protein p53 can act as a transcription factor affecting both cell cycle control and apoptosis, and we have previously shown that p53 is activated during terminal megakaryocytic (Mk) differentiation of the CHRF-288-11 (CHRF) cell line. Here, we use RNA interference to reduce p53 expression in CHRF cells and show that reduced p53 activity leads to a greater fraction of polyploid cells, higher mean and maximum ploidy, accelerated DNA synthesis, and delayed apoptosis and cell death upon phorbol 12-myristate 13-acetate-induced Mk differentiation. In contrast, reduced p53 expression did not affect the ploidy or DNA synthesis of CHRF cells in the absence of phorbol 12-myristate 13-acetate stimulation. Furthermore, primary Mk cells from cultures initiated with p53-null mouse bone marrow mononuclear cells displayed higher ploidy compared with wild-type controls. Quantitative reverse transcription-PCR analysis of p53-knockdown CHRF cells, compared with the "scrambled" control CHRF cells, revealed that six known transcriptional targets of p53 (BBC3, BAX, TP53I3, TP53INP1, MDM2, and P21) were down-regulated, whereas BCL2 expression, which is known to be negatively affected by p53, was up regulated. These studies show that the functional role of the intrinsic activation of p53 during Mk differentiation is to control polyploidization and the transition to endomitosis by impeding cell cycling and promoting apoptosis. PMID- 18397891 TI - Beta-fructofuranosidase genes of the silkworm, Bombyx mori: insights into enzymatic adaptation of B. mori to toxic alkaloids in mulberry latex. AB - Mulberry latex contains extremely high concentrations of alkaloidal sugar mimic glycosidase inhibitors, such as 1,4-dideoxy-1,4-imino-D-arabinitol (D-AB1) and 1 deoxynojirimycin (DNJ). Although these compounds do not harm the silkworm, Bombyx mori, a mulberry specialist, they are highly toxic to insects that do not normally feed on mulberry leaves. D-AB1 and DNJ are strong inhibitors of alpha glucosidases (EC 3.2.1.20); however, they do not affect the activity of beta fructofuranosidases (EC 3.2.1.26). Although alpha-glucosidase genes are found in a wide range of organisms, beta-fructofuranosidase genes have not been identified in any animals so far. In this study, we report the identification and characterization of beta-fructofuranosidase genes (BmSuc1 and BmSuc2) from B. mori. The BmSuc1 gene was highly expressed in the midgut and silk gland, whereas the expression of BmSuc2 gene was not detected. BmSuc1 encodes a functional beta fructofuranosidase, whose enzymatic activity was not inhibited by DNJ or D-AB1. We also showed that BmSUC1 protein localized within the midgut goblet cell cavities. Collectively, our data clearly demonstrated that BmSuc1 serves as a sugar-digesting enzyme in the silkworm physiology. This anomalous presence of the beta-fructofuranosidase gene in the B. mori genome may partly explain why the silkworm can circumvent the mulberry's defense system. PMID- 18397892 TI - Peak bagging for peptide mass fingerprinting. AB - MOTIVATION: Mass Spectrometry (MS)-based protein identification via peptide mass fingerprinting (PMF) is a key component in high-throughput proteome research. While PMF was the first commonly used protein identification method, provided higher throughput than the tandem MS-based method, its accuracy is lower than that of the tandem MS method. Thus, it is desirable to develop PMF-based algorithm with higher protein identification accuracy to facilitate proteome research. RESULTS: We propose a peak bagging method for single MS-based protein identification. It combines results from multiple PMF algorithms, where each PMF algorithm takes a random peak subset as input. Evaluation with a set of real MALDI-TOF MS spectra shows that the new peak bagging method provides consistent improvements over the single PMF algorithm. PMID- 18397893 TI - siRNA specificity searching incorporating mismatch tolerance data. AB - Artificially synthesized short interfering RNAs (siRNAs) are widely used in functional genomics to knock down specific target genes. One ongoing challenge is to guarantee that the siRNA does not elicit off-target effects. Initial reports suggested that siRNAs were highly sequence-specific; however, subsequent data indicates that this is not necessarily the case. It is still uncertain what level of similarity and other rules are required for an off-target effect to be observed, and scoring schemes have not been developed to look beyond simple measures such as the number of mismatches or the number of consecutive matching bases present. We created design rules for predicting the likelihood of a non specific effect and present a web server that allows the user to check the specificity of a given siRNA in a flexible manner using a combination of methods. The server finds potential off-target matches in the corresponding RefSeq database and ranks them according to a scoring system based on experimental studies of specificity. AVAILABILITY: The server is available at http://informatics-eskitis.griffith.edu.au/SpecificityServer. PMID- 18397894 TI - Domain annotation of trimeric autotransporter adhesins--daTAA. AB - MOTIVATION: Trimeric autotransporter adhesins (TAAs), such as Yersinia YadA, Neisseria NadA, Moraxella UspAs, Haemophilus Hia and Bartonella BadA, are important pathogenicity factors of proteobacteria. Their high sequence diversity and distinct mosaic-like structure lead to difficulties in the annotation of their sequences. These stem from the large number of short repeats, the presence of compositionally unusual coiled-coils, fuzzy domain boundaries and regions of seemingly low sequence complexity. RESULTS: We have developed a workflow, named daTAA, for the accurate domain annotation of TAAs. Its core consists of manually curated alignments and of knowledge-based rules that enhance assignments made by sequence similarity. Compared to general domain annotation servers such as PFAM, daTAA captures more domains and provides more sensitive domain detection, as well as integrated and detailed coiled-coil assignments. AVAILABILITY: The daTAA server is freely accessible at http://toolkit.tuebingen.mpg.de/dataa PMID- 18397895 TI - adegenet: a R package for the multivariate analysis of genetic markers. AB - The package adegenet for the R software is dedicated to the multivariate analysis of genetic markers. It extends the ade4 package of multivariate methods by implementing formal classes and functions to manipulate and analyse genetic markers. Data can be imported from common population genetics software and exported to other software and R packages. adegenet also implements standard population genetics tools along with more original approaches for spatial genetics and hybridization. AVAILABILITY: Stable version is available from CRAN: http://cran.r-project.org/mirrors.html. Development version is available from adegenet website: http://adegenet.r-forge.r-project.org/. Both versions can be installed directly from R. adegenet is distributed under the GNU General Public Licence (v.2). PMID- 18397896 TI - Expression of telomeric repeat binding factor 1 protein in nonsmall cell lung cancer with human telomerase reverse transcriptase positive. AB - Telomeric repeat binding factor 1 plays pivotal roles in telomere protection and maintenance in mammalian cells. In this article, the expression of telomeric repeat binding factor 1 protein in nonsmall cell lung cancer with human telomerase reverse transcriptase positive is investigated, and the relationship between the telomeric repeat binding factor 1 and clinic factors is analyzed. The expressions of human telomerase reverse transcriptase were detected by immunohistochemistry. The levels of telomeric repeat binding factor 1 protein were measured by Western blot. In all, 72% (36/50) patients showed human telomerase reverse transcriptase positive. Telomeric repeat binding factor 1 expression of cancer tissues and paired noncancerous tissues in 30 cases of human telomerase reverse transcriptase positive was 0.552 +/- 0.329 and 0.654 +/- 0.476, respectively (P < .05). No significant difference in telomeric repeat binding factor 1 protein expression was observed among sex, clinical stages, pathological subtypes, and lymph node metastasis. The results indicated that down regulation of telomeric repeat binding factor 1 expression appeared in lung cancer tissue, and no correlation was found between telomeric repeat binding factor 1 expression and clinicopathologic factors. PMID- 18397897 TI - Double synchronous pulmonary lymphatic-related lesions. AB - A wide range of pathologies may primarily affect the lymphatic vessels in the lungs. In this article, a unique case of pulmonary silicosis associated with a subtle lymphangitic carcinomatosis from an unknown prostate cancer is reported and discussed. PMID- 18397898 TI - Cystic struma ovarii (with macrocystic change). AB - In this article, a case of cystic struma ovarii with macrocystic change is presented. Cysts derived from struma ovarii may mimic a mucinous or serous cystadenoma. A careful examination for any thyroid microfollicles within fibrous septa or areas of solid stroma is key. In problematic cases, immunohistochemical staining with thyroglobulin and thyroid transcription factor-1 may be required to establish the diagnosis. PMID- 18397899 TI - Ovarian adenomyoma mimicking an ovarian malignancy: a case report with literature review. AB - Smooth muscle tumors of the ovary are rare, and ovarian adenomyoma are even rarer. It is a well-circumscribed biphasic tumor composed of benign glands and smooth muscle cells, as well as a variety of pseudoneoplastic glandular lesion. After extensive literature search, the case presented in this article appears to be the third case of an ovarian adenomyoma and the first case in a postmenopausal woman. PMID- 18397900 TI - Metaplastic thymoma: report of an unusual thymic epithelial neoplasm arising in the wall of a thymic cyst. AB - In this report, a case of metaplastic thymoma arising in a thymic cyst wall is discussed. The patient was a 61-year-old male whose chief complaint was sweating and chest pain. Imaging study revealed a mediastinal mass with right hemithoracic extension. He underwent surgical resection of the mass and remnants of the thymus. Histologic studies showed a primary thymic neoplasm with a biphasic histologic pattern composed of 2 distinct epithelial and stromal components arising in a thymic cyst wall. Immunohistochemically, the epithelial component was cytokeratin positive and focally reactive for EMA. Marked expression of EMA and vimentin was seen in spindle cells. Metaplastic thymoma is an extremely rare variant of primary thymic epithelial neoplasms with only a few cases reported in the literature. PMID- 18397901 TI - Immunoprofile of kuttner tumor (chronic sclerosing sialadenitis). AB - In the present study, the immunoprofile of chronic sclerosing sialadenitis, also known as Kuttner tumor, was analyzed. Two cases that occurred in the submandibular gland of male patients were submitted to immunohistochemical reactions to different antibodies. Histological examinations showed a submandibular gland exhibiting various degrees of atrophy with destruction of acini, infiltration by inflammatory cells, and periductal fibrosis. Reactions to cytokeratins (CKs) showed acini and duct remnants positive to CKs 7, 8, 19, and 13. CK14 stained myoepithelial cells around preserved acini and intercalated duct, and also basal cell of excretory ducts, but was negative in proliferating and branching ducts. Smooth muscle actin (SMA) was expressed by myofibroblasts in periductal fibrosis, and an intense expression of extracellular components was also seen. Lymphocyte markers showed, besides mature follicles, a higher presence of CD45RO positive cells. Thus, the immunoprofile of Kuttner is much more in keeping with an inflammatory-induced degenerative disease than with a preneoplastic lesion. PMID- 18397902 TI - Pigmented oral squamous cell carcinoma: a case report and brief review of the literature. AB - Melanin impregnation in squamous cell carcinoma (SCC) is an uncommon histological finding. Rare nonmelanocytic entities were previously described as having melanocyte colonization. A 57-year-old Brazilian woman was referred with a pigmented lesion in the lower lip and alveolar ridge with a prior clinical diagnosis of melanoma. The incisional biopsy of the tumor revealed an SCC with strong colonization of melanocytes in the stroma. The authors report a case of an unusual SCC variant and a brief review of the literature. PMID- 18397903 TI - Development and implementation of a pediatric cardiac anesthesia/intensive care database. AB - Systematic collection and electronic storage of data can assist in improving quality and efficiency of patient care and can provide a data set to interrogate for subsequent performance improvement and clinical research purposes. In this article, an electronic perioperative pediatric cardiac surgery database to be used by a multidisciplinary care team was designed, developed, and implemented. Technical goals for the design included low cost, rapid development and implementation, adequate security, and potential for internal and external distribution. Implementation of the described database has proved to be invaluable for quality assurance and statistical analysis of data relevant to patient care. From the overall positive experience, it was concluded that the electronic data management does not always need major cost investment. PMID- 18397905 TI - Recent advances in application of cerebral oximetry in adult cardiovascular surgery. AB - Cerebral oximetry is a noninvasive technology that continuously monitors cerebral tissue oxygen saturation, which is a sensitive index of global cerebral hypoperfusion. On the basis of near-infrared spectroscopy technology, information is provided on the availability of oxygen in brain tissue at risk during numerous pathological conditions. Complementary to the arterial oxygen saturation measured by pulse oximetry, cerebral tissue oxygen saturation reflects regional cerebral metabolism and the balance of local cerebral oxygen supply/demand. Recently, patient management with the guidance of cerebral oximetry has resulted in improved patient outcomes. This review will briefly describe the physics behind cerebral oximetry and will provide an overview of the literature focusing primarily on articles published within the past 5 years. PMID- 18397904 TI - Anesthetic considerations in the patient with valvular heart disease undergoing noncardiac surgery. AB - Valvular heart disease can be an important finding in patients presenting for noncardiac surgery. Valvular heart disease and resulting comorbidity, such as heart failure or atrial fibrillation, significantly increase the risk for perioperative adverse events. Appropriate preoperative assessment, adequate perioperative monitoring, and early intervention, should hemodynamic disturbances occur, may help prevent adverse events and improve patient outcome. This review article aims to guide the practitioner in the various aspects of anesthetic management in the perioperative care of patients with valvular heart disease. The pharmacological approach to optimization of patient outcome with drugs, such as betablockers and lipid-lowering medications (statins), is an evolving field, and recent developments are discussed in this article. PMID- 18397906 TI - Cardiac anesthesia and surgery in geriatric patients. AB - The average age of US population is steadily increasing, with more than 15 million people aged 80 and older. Coronary artery disease and degenerative cardiovascular diseases are particularly prevalent in this population. Consequently, an increasing number of elderly patients are referred for surgical intervention. Advanced age is associated with decreased physiologic reserve and significant comorbidity. Thorough preoperative assessment, identification of the risk factors for perioperative morbidity and mortality, and optimal preparation are critical in these patients. Age-related changes in comorbidities and altered pharmacokinetics and pharmacodynamics impacts anesthetic management, perioperative monitoring, postoperative care, and outcome. This article updates the age-related changes in organ subsystems relevant to cardiac anesthesia, perioperative issues, and intraoperative management. Early and late operative outcome in octogenarians undergoing cardiac surgery are reviewed. The data clearly indicate that no patient group is "too old" for cardiac surgery and that excellent outcomes can be achieved in selected group of elderly patients. PMID- 18397907 TI - Hazardous off-gassing of carbon monoxide and oxygen depletion during ocean transportation of wood pellets. AB - Five ocean vessels were investigated for the characterization and quantification of gaseous compounds emitted during ocean transportation of wood pellets in closed cargo hatches from Canada to Sweden. The study was initiated after a fatal accident with several injured during discharge in Sweden. The objective with the investigation was to better understand the off-gassing and issues related to workers' exposure. Air sampling was done during transport and immediately before discharge in the undisturbed headspace air above the wood pellets and in the staircase adjacent to each hatch. The samples were analyzed with Fourier transform infrared spectroscopy and direct reading instruments. The following compounds and ranges were detected in samples from the five ships: carbon monoxide (CO) 1460-14650 ppm, carbon dioxide (CO2) 2960-21570 ppm, methane 79.9 956 ppm, butane equivalents 63-842 ppm, ethylene 2-21.2 ppm, propylene 5.3-36 ppm, ethane 0-25 ppm and aldehydes 2.3-35 ppm. The oxygen levels were between 0.8 and 16.9%. The concentrations in the staircases were almost as high as in the cargo hatches, indicating a fairly free passage of air between the two spaces. A potentially dangerous atmosphere was reached within a week from loading. The conclusions are that ocean transportation of wood pellets in confined spaces may produce an oxygen deficient atmosphere and lethal levels of CO which may leak into adjacent access spaces. The dangerous combination of extremely high levels of CO and reduced oxygen produces a fast-acting toxic combination. Measurement of CO in combination with oxygen is essential prior to entry in spaces having air communication with cargo hatches of wood pellets. Forced ventilation of staircases prior to entry is necessary. Redesign, locking and labeling of access doors and the establishment of rigorous entry procedures and training of onboard crew as well as personnel boarding ocean vessels are also important. PMID- 18397908 TI - Cross-talk among Toll-like receptors and their ligands. AB - Toll-like receptors (TLRs) 4, 5, 7 and 9 belong to a family of proteins that recognize mainly conserved microbial motifs. Though each TLR has a highly specific ability to recognize a particular microbial pattern, recent papers suggest that some ligands are able to affect the expression of different TLRs. In this paper, we have investigated TLR4, 5, 7 and 9 expression, both at mRNA and protein level, following treatment of different intestinal epithelial cell lines with LPS, flagellin, loxiribine, CpG-oligodeoxynucleotide and peptidoglycan, to assess if the different TLR ligands may modulate the expression of the respective TLR and of the unrelated ones. Our results show that a cross-talk exists between TLRs and various ligands, indicating a cross-regulation among these pattern recognition receptors. In particular, TLR4 was generally down-regulated by treatment with ligands other than LPS, while flagellin and unrelated microbial associated molecular patterns exerted a general stimulatory activity as regards TLR5 expression. Concerning TLR7 and 9, we have observed a more variable behaviour of the various cell lines with the different ligands. Together, our results demonstrate that the expression of TLRs in intestinal cells is highly dynamic and tightly regulated in response to encountered microbial stimuli. PMID- 18397909 TI - Improved methods for detecting selection by mutation analysis of Ig V region sequences. AB - Statistical methods based on the relative frequency of replacement mutations in B lymphocyte Ig V region sequences have been widely used to detect the forces of selection that shape the B cell repertoire. However, current methods produce an unexpectedly high frequency of false positives and are sensitive to intrinsic biases of somatic hypermutation that can give the appearance of selection. The new statistical test proposed here provides a better trade-off between sensitivity and specificity compared with previous approaches. The low specificity of existing methods was shown in silico to result from an interaction between the effects of positive and negative selection. False detection of positive selection was confirmed in vivo through a re-analysis of published sequence data from diffuse large B cell lymphomas, highlighting the need for re analysis of some existing studies. The sensitivity of the proposed method to detect selection was validated using new Ig transgenic mouse models in which positive selection was expected to be a significant force, as well as with a simulation-based approach. Previous concerns that intrinsic biases of somatic hypermutation could give the appearance of selection were addressed by extending the current mutation models to more fully account for the impact of microsequence on relative mutability and to include transition bias. High specificity was confirmed using a large set of non-productively rearranged Ig sequences. These results show that selection can be detected in vivo with high specificity using the new method proposed here, allowing greater insight into the existence and direction of antigen-driven selection. PMID- 18397910 TI - Osteoclasts support the survival of human plasma cells in vitro. AB - The aim of this in vitro study was to evaluate if osteoclasts (OCs) and dendritic cells (DCs), both of monocyte origin, can support the survival of normal human plasma cells (PCs). PCs differentiate from plasmablasts (PBs) arising from activated B cells, essentially memory B cells. To study the survival of both PBs (CD20(low)CD38(high)CD138(neg)) and PCs (CD20(neg)CD38(bright)CD138(bright)), we generated pre-PBs (CD20(low)CD38(pos)CD138(neg)) from CD40-activated B cells (CD20(high)CD38(neg)CD138(neg)) and cultured them on DCs or OCs in the presence of added IL-6. By quantitative and qualitative study, we showed that DCs support the survival of PBs and early PCs, but not that of PCs. In contrast, OCs support the survival of PBs, early PCs and PCs. PCs surviving on OCs 12 days after pre-PB input display phenotypic features of bone marrow PCs, CD138(bright)CD38(bright)HLA-DR(low)CD45(dim). The ability for OCs to support the survival of PCs was fully dependent on cell-cell contact and not inhibited by BCMA-Fc suggesting that secreted BAFF and APRIL were not involved. PMID- 18397911 TI - Severe pharyngitis in stockbreeders: an unusual presentation of brucellosis. AB - Abstract Brucellosis is a known occupational hazard for shepherds, abattoir workers, veterinarians, dairy industry professionals and personnel in microbiological laboratories. Any organ may be affected by Brucella species but to date, severe manifestations in the pharynx have never been reported as the prevailing features of brucellosis. We report two cases in stockbreeders who presented with high-grade fever and severe exudative pharyngitis accompanied by severe odynophagia in the first and with high-grade fever and a history of relapsing tonsillitis in the second. We therefore recommend including brucellosis in the differential diagnosis of febrile patients suffering from unexplained pharyngitis or tonsillitis who belong to high-risk groups for contracting brucellosis. PMID- 18397912 TI - Absence of platinum salt sensitivity in autocatalyst workers exposed to tetraamine platinum dichloride. AB - BACKGROUND: Platinum salt sensitivity (PSS) is well recognized following occupational exposure to platinum salts, though specific platinum compounds have been suggested to be non-allergenic. We report on a cohort of autocatalyst workers exposed to tetraamine platinum dichloride (TPC) and other platinum-group elements. METHODS: All subjects employed at an autocatalyst production plant undertook medical surveillance with symptoms, examination findings and results of skin prick testing and spirometry prospectively recorded. Environmental testing of the workplace was also performed to determine the level of exposure. RESULTS: Twenty-six subjects had a mean duration of employment of 46 (+/-30) months and undertook a mean 6.8 (+/-4.3) examinations. No subjects described the development of new respiratory or dermatological symptoms. No patients developed positive skin reactivity to platinum salts. FEV(1) remained unchanged for all subjects over the course of the study period. CONCLUSIONS: TPC and platinum-group elements are not associated with the development of PSS or occupational asthma. Identification of chemical compounds is important when advising on occupational health screening. TPC and/or platinum-group elements should be used in preference to chloroplatinic acid in catalyst production to minimize the impact of occupational illness due to PSS. PMID- 18397913 TI - Toenail nicotine levels as predictors of coronary heart disease among women. AB - The authors assess the ability of toenail nicotine levels as a biomarker to predict incident coronary heart disease (CHD). A nested case-control study was carried out among 62,641 women aged 36-61 years in the Nurses' Health Study cohort who provided toenail clippings in 1982. Between 1984 and 1998, 905 incident CHD cases were diagnosed and matched with two controls by age and date of toenail collection. Using multivariate logistic regression analyses, the authors found a statistically significant dose-response association between increasing toenail nicotine levels and risk of CHD (p(trend) < 0.0001); women in the highest quintile had a relative risk of 3.44 (95% confidence interval (CI): 2.56, 4.62) compared with women in the lowest quintile. With each increase in the log-transformed unit of continuous toenail nicotine levels, there was a 42% increase in the risk of CHD (relative risk = 1.42, 95% CI: 1.33, 1.52). The association remained significant when the number of cigarettes smoked and passive smoking were included as covariates (relative risk = 1.12, 95% CI: 1.01, 1.24). In conclusion, toenail nicotine levels are predictive of CHD among women independent of other risk factors and remained significant even after adjustment for history of cigarette smoking. PMID- 18397915 TI - Suppression of the steroid-primed luteinizing hormone surge in the female rat by sodium dimethyldithiocarbamate: relationship to hypothalamic catecholamines and GnRH neuronal activation. AB - In female rodents, hypothalamic norepinephrine (NE) has a role in stimulating the secretion of gonadotropin-releasing hormone (GnRH) that triggers the ovulatory surge of luteinizing hormone (LH). NE synthesis from dopamine (DA) is catalyzed by dopamine-beta-hydroxylase (DbetaH) which contains a copper cofactor. Sodium dimethyldithiocarbamate (DMDC) is a pesticide with metal chelating properties that has been found to reduce DbetaH activity. The resultant decrease in NE causes a suppression of both the LH surge and ovulation. The present study examined the dose-related impact of DMDC on hypothalamic GnRH neuronal activation indicated by the nuclear presence of the early gene product c-fos. It represents an essential link between effects on NE and suppression of the surge. Ovariectomized (OVX), estradiol-, and progesterone-primed Sprague-Dawley rats were given a single ip injection of 0, 3.6, 7.1, 14.2, or 28.4 mg/kg DMDC in separate groups of females to assess tissue GnRH/c-fos immunostaining, hypothalamic catecholamines, and serial blood samplings for LH. A dose-related decline in hypothalamic NE and increase in DA at 2 h after DMDC administration were consistent with a decrease in c-fos-positive GnRH neurons, with an almost complete absence of c-fos at the two highest doses. The effects correlated well with a suppression of the surge, although the percentage decrease in c-fos neurons at 7.1 mg/kg only attenuated the surge peak, not the overall amount of circulating LH. The present data offer further evidence that the impact of DMDC on the LH surge is central in origin and in doing so defines the toxic pathway for this effect on ovulation. PMID- 18397914 TI - The Shanghai Women's Asthma and Allergy Study: objectives, design, and recruitment results. AB - The Shanghai Women's Asthma and Allergy Study is the first population-based incidence study designed to assess the associations of dietary antioxidant intake and measures of oxidative stress and antioxidant enzyme activity with development of adult-onset asthma and allergic rhinitis. A total of 65,732 participants in the Shanghai Women's Health Study, an ongoing cohort study in seven districts of Shanghai, People's Republic of China, were recruited to the Shanghai Women's Asthma and Allergy Study from 2003 to 2007. Dietary intake was assessed in the parent study by using a validated and quantitative food frequency questionnaire at baseline recruitment and at the first biennial follow-up survey. Blood and urine samples were collected to measure baseline oxidative stress, antioxidant enzyme activity, and nutrient levels at the baseline survey. Incident asthma and allergic rhinitis were assessed by using a modification of the International Study of Asthma and Allergies in Childhood questionnaire during the biennial in person survey of the Shanghai Women's Health Study. Diagnosis of asthma was confirmed by either methacholine challenge testing or test of reversibility to beta-agonists. Dietary antioxidant intake, plasma antioxidants, antioxidant enzymes, and urinary isoprostanes, a marker of oxidative stress, were measured prior to disease onset. This paper describes the study objectives, design, population demographics, and recruitment results. PMID- 18397916 TI - Effects of a 3beta-hydroxysteroid dehydrogenase inhibitor, trilostane, on the fathead minnow reproductive axis. AB - A number of environmental contaminants and plant flavonoid compounds have been shown to inhibit the activity of 3beta-hydroxysteroid dehydrogenase/Delta(5) Delta(4) isomerase (3beta-HSD). Because 3beta-HSD plays a critical role in steroid hormone synthesis, inhibition of 3beta-HSD represents a potentially important mode of endocrine disruption that may cause reproductive dysfunction in fish or other vertebrates. The objective of this study was to test the hypothesis that exposure to the model 3beta-HSD inhibitor, trilostane, would adversely affect reproductive success of the fathead minnow (Pimephales promelas). Results of in vitro experiments with fathead minnow ovary tissue demonstrated that trilostane inhibited 17beta-estradiol (E2) production in a concentration- and time-dependent manner, and that the effect was eliminated by providing a substrate (progesterone) that does not require 3beta-HSD activity for conversion to E2. Exposure of fish to trilostane caused a significant reduction in spawning frequency and reduced cumulative egg production over the course of the 21-day test. In females, exposure to 1500 mug trilostane/l reduced plasma vitellogenin concentrations, but did not cause significant histological alterations. In males, average trilostane concentrations as low as 50 mug/l significantly increased testis mass and gonadal somatic index. Trilostane exposure did not influence the abundance of mRNA transcripts coding for 3beta-HSD or other steroidogenesis regulating proteins in males or females. As a whole, results of this study support the hypothesis that 3beta-HSD inhibition can cause reproductive dysfunction in fish, but did not yield a clear profile of responses at multiple levels of biological organization that could be used to diagnose this mode of action. PMID- 18397917 TI - Evidence that the anticarcinogenic effect of caffeic acid phenethyl ester in the resistant hepatocyte model involves modifications of cytochrome P450. AB - Caffeic acid phenethyl ester (CAPE), a natural component of propolis, shows anticarcinogenic properties in the modified resistant hepatocyte model when administered before initiation or promotion of hepatocarcinogenesis process; however, information about the mechanism underlying this chemoprotection is limited. The aim of this work was to characterize the effect of CAPE on cytochrome P450 (CYP), which is involved in diethylnitrosamine (DEN) metabolism during the initiation stage of chemical hepatocarcinogenesis. Male Fischer-344 rats were treated as in the modified resistant hepatocyte model. Liver samples were obtained at four different times: at 12 h after pretreatment with CAPE and at 12 and 24 h and 25 days after DEN administration. Liver damage was determined by histology with hematoxylin and eosin, measurement of total CYP levels and enzyme activity, and gamma-glutamyl transpeptidase-positive (GGT+) staining of hepatocyte foci. CAPE administration prevented DEN-induced necrosis at 24 h. It also decreased O-dealkylation of 7-ethoxy-resorufin (EROD), O-dealkylation of 7 methoxyresorufin (MROD), and 7-pentoxy-resorufin activities at 12 h after its administration and EROD and MROD activities at 12 h after administration of DEN. CAPE treatment decreased GGT+ foci by 59% on day 25. Our results suggest that CAPE modifies the enzymatic activity of CYP isoforms involved in the activation of DEN, such as CYP1A1/2 and CYP2B1/2. These findings describe an alternative mechanism for understanding the ability of CAPE to protect against chemical hepatocarcinogenesis. PMID- 18397918 TI - Why does ARNT2 behave differently from ARNT? PMID- 18397919 TI - jModelTest: phylogenetic model averaging. AB - jModelTest is a new program for the statistical selection of models of nucleotide substitution based on "Phyml" (Guindon and Gascuel 2003. A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol. 52:696-704.). It implements 5 different selection strategies, including "hierarchical and dynamical likelihood ratio tests," the "Akaike information criterion," the "Bayesian information criterion," and a "decision-theoretic performance-based" approach. This program also calculates the relative importance and model-averaged estimates of substitution parameters, including a model averaged estimate of the phylogeny. jModelTest is written in Java and runs under Mac OSX, Windows, and Unix systems with a Java Runtime Environment installed. The program, including documentation, can be freely downloaded from the software section at http://darwin.uvigo.es. PMID- 18397920 TI - Pain and the immune system. AB - In inflammation, leucocytes containing opioid peptides migrate into the tissue. Opioid peptides can be released and bind to opioid receptors on peripheral nerve terminals, which counteracts inflammatory pain. Migration of opioid peptide containing leucocytes is controlled by chemokines and adhesion molecules. Neurokinins, such as, substance P also contribute to the recruitment of these cells. Opioid peptide release from granulocytes can be stimulated by chemokines, such as, CXCR2 ligands. The release is dependent on intracellular calcium and activation of phosphoinositol-3 kinase and p38 mitogen activated kinase. Endogenous opioid peptides produced by leucocytes not only confer analgesia but recent evidence supports the concept that they also prevent the development of tolerance at peripheral opioid receptors. This review presents the discoveries that led to the concept of analgesia produced by immune-derived opioids. PMID- 18397921 TI - Incidence of postoperative nausea and emetic episodes after xenon anaesthesia compared with propofol-based anaesthesia. AB - BACKGROUND: Xenon has been proved to be safe and efficacious for general anaesthesia in numerous trials. In addition, experimental studies demonstrate that xenon inhibits the 5-hydroxytryptamine type 3 (5-HT(3)) receptor. As 5-HT(3) receptor antagonists are known to decrease postoperative nausea and vomiting (PONV) to an extent comparable with a propofol-based total i.v. technique, we tested the hypothesis that general anaesthesia with xenon would result in a reduced incidence of PONV similar to that observed with propofol-based anaesthesia. METHODS: After obtaining approval from the local ethics committee and written informed consent, 142 patients were randomized to receive xenon anaesthesia or propofol-based total i.v. anaesthesia (TIVA), both supplemented with remifentanil. The incidence of postoperative nausea and emetic episodes was recorded in the post-anaesthesia care unit and on the ward more than 24 h after anaesthesia. RESULTS: A total of 142 patients were equally distributed between the xenon and TIVA groups. Anaesthesia was maintained with mean (sd) concentrations of either xenon 61 (2)% or propofol 100 (20) microg kg(-1) min( 1). Incidences of nausea and emetic episodes over the whole 24-h period were 66.2% and 35.2% in the xenon group and 26.8% and 16.9% in the TIVA group (P<0.001 and P<0.021). CONCLUSION: Despite knowing the 5-HT(3) antagonistic properties of xenon, its use is associated with a higher incidence of nausea and emetic episodes compared with TIVA with propofol. PMID- 18397922 TI - Endogenous antimicrobial peptide LL-37 induces human vasodilatation. AB - BACKGROUND: Septic shock includes blood vessel dilatation and activation of innate immunity, which in turn causes release of antimicrobial peptides such as LL-37. It has been shown that LL-37 can attract leucocytes via the lipoxin A(4) receptor (ALX, FPRL1). ALX is also present in vascular endothelial cells. To explore possible ways of pharmacological intervention in septic shock, we investigated if LL-37 can affect vascular tone. METHODS: Human omental arteries and veins were obtained during abdominal surgery, and circular smooth muscle activity was studied in organ baths. Gene expression was studied using reverse transcriptase-polymerase chain reaction. RESULTS: LL-37, at micromolar concentrations, induced a concentration- and endothelium-dependent relaxation in vein but not in artery segments precontracted by endothelin-1. The relaxation was profoundly reduced by potassium chloride (30 mM) to inhibit endothelium-derived hyperpolarizing factor (EDHF), whereas it was less affected by the NOS inhibitor, l-N(G)-nitroarginine methyl ester, and not at all by indomethacin. The ALX agonist, WKYMVm, also induced a relaxation and both the relaxations induced by LL 37 and WKYMVm were inhibited by the ALX antagonist, WRWWWW. ALX was expressed in the vein endothelium. CONCLUSIONS: We demonstrate, for the first time, that the human antimicrobial peptide, LL-37, induces endothelium-dependent relaxation in human omental veins mediated via an effect on endothelial ALX. The relaxation involves the release of nitric oxide and EDHF but not prostanoids. LL-37 released from white blood cells could contribute to blood vessel dilatation during sepsis and treatment with ALX antagonists might be successful. PMID- 18397923 TI - Susceptibility of Staphylococcus aureus isolated from skin and wound infections in the United States 2005-07: laboratory-based surveillance study. AB - OBJECTIVES: The aim of this study was to describe the rates of antimicrobial susceptibility of Staphylococcus aureus from skin and wound infections reported from nine regions of the USA during 2005-07 and to identify the regional variation in patterns of resistance. METHODS: The Surveillance Network (TSN) comprises 296 laboratories across the nine census regions of the USA. TSN laboratories reported the susceptibility data for six antimicrobials by isolate with source and other relevant data. Antimicrobial susceptibility data were analysed by individual drug resistance, multidrug resistance and geographical distribution of resistance phenotypes. RESULTS: There were over 380 000 isolates of S. aureus tested and reported for the period 2005-07. Methicillin resistance was observed in 57.8% in 2007, with little change from 2005. There was little difference in rates of methicillin resistance between community and hospital strains, although strains from intensive care units (ICUs) tended to be slightly more resistant overall. Resistance to other antimicrobials was also reported. A regional variation in resistance rates was noted with the highest rates in the Central states and lowest in the New England and Mid-Atlantic regions. There was high activity observed with trimethoprim/sulfamethoxazole and gentamicin. Linezolid resistance was rare. Oxacillin resistance was similar among paediatric and elderly cohorts, whereas ciprofloxacin and clindamycin resistance was significantly (P < 0.01) more common in elderly patients when compared with both paediatric and adult populations. Less than a third of all isolates showed no resistance mechanism, 30.3%. Three distinct resistance phenotypes accounted for 46% of all resistant strains. Overall, there were more highly drug-resistant isolates from the ICU with four, five or six drug-resistant phenotypes accounting for over a third of all strains. CONCLUSIONS: S. aureus has become methicillin resistant in both the community and hospital settings; however, little change has been seen in the past 3 years. Multiresistant strains now are seen in all settings, but due to regional variation, empirical therapy should be guided by local susceptibility patterns. Currently, among the agents studied, only trimethoprim/sulfamethoxazole, gentamicin and linezolid exhibit susceptibility rates of >95%. PMID- 18397924 TI - Susceptibility of 71 French isolates of Francisella tularensis subsp. holarctica to eight antibiotics and accuracy of the Etest method. PMID- 18397925 TI - Opposing expectations and suboptimal use of a local antibiotic hospital guideline: a qualitative study. AB - OBJECTIVES: The aim of this study was to determine the opinions and problems concerning the use of a local antibiotic hospital guideline in a 1900-bed tertiary-care, university teaching hospital. METHODS: A qualitative study using focus group discussions explored the usability and applicability of local antibiotic guidelines together with possible supportive measures. The sample included 22 physicians, deliberately divided between internal medicine (59.1%) and surgery (40.9%), and levels of experience (59.1% residents; 40.9% supervisors). Focus groups were conducted within one specific subgroup. Analysis was carried out using a framework analysis approach. RESULTS: General acceptance of local guidelines was high but clear differences were present between subgroups with different desires and requirements from guideline contents. Opposing views were present towards supportive measures, especially multidisciplinary collaboration. Guideline distribution and accessibility appeared to be confusing, resulting in delayed application. An important supplementary barrier was the need to collect the guideline personally. Supervisors in their role as opinion leaders were mentioned as highly influential towards residents' practice. CONCLUSIONS: Locally developed hospital guidelines experience the same barriers as other guidelines. Within one hospital, prescribers have to be seen as a number of different target groups instead of a homogeneous population. For an optimal effect, interventions will have to consider these differences. Also, in order to improve local guideline use and antibiotic consumption, supervisors have to be aware of how their role as opinion leaders can influence residents. Lastly, active guideline distribution and promotion remains critical to ensure efficient guideline use. Future research should focus on how to adapt interventions to these different target groups. PMID- 18397926 TI - Assessment of the dose from radon and its decay products in the Bozkov dolomite cave. AB - The dose from radon and its progeny remains a frequently discussed problem. ICRP 65 provides a commonly used methodology to calculate the dose from radon. Our work focuses on a cave environment and on assessing the doses in public open caves. The differences in conditions (aerosol size distribution, humidity, radon and its progeny ratio, etc.) are described by the so-called cave factor j. The cave factor is used to correct the dose for workers which is calculated using the ICRP 65 recommendation. In this work, the authors have brought together measured data of aerosol size distribution, unattached and attached fraction activity, and have calculated the so-called cave factor for the Bozkov dolomite cave environment. The dose conversion factors based on measured data and used for evaluating the cave factor were calculated by LUDEP software, which implements HRTM ICRP66. PMID- 18397927 TI - Soil gas radon, indoor radon and gamma dose rate in CZ: contribution to geostatistical methods for European atlas of natural radiations. AB - A comparison of Czech indoor radon data, soil gas radon data and gamma dose rate was performed on the data sets of 92,276 indoor radon measurements in existing dwellings (National Radiation Protection Institute - NRPI), database of 9500 test sites of soil gas radon measurements (Czech Geological Survey - CGS) georeferenced to levels of gamma dose rate map. Three methods were used for the study of soil gas Rn-indoor Rn relationship: (1) based on the vectorised point soil gas and indoor data related to vectorised areas of gammadose rate, (2) vectorised soil gas-indoor data based on vectorised geological units and (3) soil gas and indoor data related to grid squares 10 x 10 km2. The first and second methods seem to express the closer correlation compared with the third one, however the correlation using the third method is influenced by the representativeness of data value in the square of 100 km2 area. On the other hand, the third method can be used for the overview coverage of the continental areas with lack of input information. PMID- 18397928 TI - New technique for the determination of radon diffusion coefficient in radon-proof membranes. AB - This paper describes a new device and a method to determine the radon diffusion coefficient in damp-proof membranes developed in the Czech Republic. The main advantage of the device is that it enables tests to be carried out in all the known measuring modes used throughout Europe. Two recently developed computer programs are presented for the numerical modelling of the time-dependent radon transport through damp-proof membranes. According to this method, the radon diffusion coefficient is derived from the process of fitting the numerical solution to the measured curve of radon concentration in a receiver container. Numerical simulation and measured data are also compared. Reasons for disagreements between different methods and specific configurations of the measuring device are also discussed. PMID- 18397929 TI - Mitigation of ineffective measures against radon. AB - Reasons of low effectiveness of radon remedial measures have been studied on several unsuccessfully remediated houses. Based on the thorough analysis, factors responsible for failures are clarified. The possibilities of how to improve the effectiveness of installed measures are also discussed. Experience in this field is documented by several examples of additionally mitigated houses. After application of additional measures, indoor radon concentration in the studied houses decreased in average 5.3 times. Costs for additional mitigation were at least four times cheaper compared with costs required for installation of original measures. PMID- 18397930 TI - Dose conversion of radon exposure according to new epidemiological findings. AB - In 1993, ICRP-65 recommended that dose conversion of radon exposure should be based on the comparison of detriments between radon exposure and effective dose. The lifetime detriment from the radon exposure was projected according to the epidemiological studies of uranium miners then available. The projection model (GSF) was multiplicative with temporal and age-at-exposure modification. Since 1993, new studies of uranium miners have appeared and many original studies were updated. In addition, projections of the risk have been improved by including further modifying factors as for instance in BEIR VI. New analyses were completed in the Czech and French studies of uranium miners with accurate estimates of exposures based on extensive radon measurements. The resulting estimates of excess absolute lifetime risk per unit exposure in working level months (WLM) from these models lead to dose conversion of 10 mSv WLM(-1) for the BEIR VI model and 8 mSv WLM(-1) for the joint Czech-French model in contrast to the conversion of 5 mSv WLM(-1) for the GSF model. PMID- 18397931 TI - Development of training syllabi for radiation protection and quality assurance of dual-energy X-ray absorptiometry (DXA) systems. AB - The use of dual-energy X-ray absorptiometry (DXA) scanners for measuring bone mineral density (BMD) is on the increase. A single DXA scan is a relatively low dose diagnostic X-ray examination; however, radiation protection (RP) issues should not be trivialised. One objective of the EU 6th Framework SENTINEL co ordination action was to develop training syllabi in RP and quality assurance (QA) for BMD, and this study presents the results. An EU-wide survey was carried out which confirmed that there was a need for an accredited DXA RP training course in many EU states. There is also limited published guidance on acceptance testing/QA for DXA. Two training syllabi were developed: one on RP and one on QA of DXA systems. A training course was delivered in Ireland in 2006 by the Medical Physics & Bioengineering Department of St James's Hospital, Dublin. Following the training course, a PC-based training CD was developed and will be made available. A harmonised approach to training will promote consistent approaches to radiation safety across the EU. PMID- 18397932 TI - Corrections to air kerma and exposure measured with free air ionisation chambers for charge of photoelectrons, Compton electrons and Auger electrons. AB - The signal charge from a free air ionisation chamber for the measurement of air kerma and exposure consists of not only the charge of ion pairs produced by secondary electrons (i.e. photoelectrons, Compton electrons and Auger electrons), but also the charge of the secondary electrons and single and multiple charged ions formed by the release of the secondary electrons. In the present work, correction factors for air kerma and exposure for the charge of the secondary electrons and ions were calculated for photons with energies in the range from 1 to 400 keV. The effects of an increase in the W value of air for low-energy electrons were also taken into consideration. It was found that the correction factors for air kerma and exposure have a maximum value near a photon energy of 30 keV; in the lower energy region, the correction factor for exposure monotonically decreases with a decrease in photon energy except for a small dip due to K-edge absorption by argon atoms in air. The values of the correction factors were found to be 0.9951 and 0.9892, respectively, for a spectrum with a mean energy of 7.5 keV, the reference X-ray spectrum with the lowest mean energy in ISO 4037-1. The air kerma correction is smaller than that for exposure, because for air kerma the signal due to the charge of secondary electrons and ions is partly compensated by the decrease in the number of ion pairs produced by the secondary electrons due to the increase of the W value of air for lower energy electrons. PMID- 18397933 TI - Rescue peri-operative management of the patient with giant electrical storm and severe left ventricular dysfunction: support by levosimendan and intraaortic balloon counterpulsation. AB - We present a case of a 62-year-old male patient (coronary heart disease, and stenosis of aortic valve) with severe left ventricular dysfunction (left ventricular ejection fraction 20%, left ventricular end-diastolic diameter 80 mm, end-diastolic volume 329 ml) who developed giant electrical storm (on the whole 115 episodes of pulseless ventricular tachycardia treated by antiarrhythmics and electrical discharges) with low-elevation of cardiac biomarkers. The patient was referred to emergent cardiac surgery (double coronary bypass grafting, aortic valve replacement, and implantation of left ventricular epicardial electrode). Levosimendan and intraaortic balloon counterpulsation were used for successful weaning from the cardiopulmonary bypass; no other arrhythmia appeared in the post bypass period. Postoperatively no arrhythmic events were detected, and repeated echocardiographic examinations of the patient in good general condition showed gradual improvement of left ventricular ejection fraction (30-35%), likewise in other parameters (left ventricular end-diastolic diameter 72 mm, end-diastolic volume 285 ml). PMID- 18397934 TI - A living model of managed competition: a conversation with Dutch health minister Ab Klink. Interview by Alain Enthoven. AB - As of 2006, the Netherlands requires all citizens to buy a standard package of health benefits from private insurers. The government subsidizes premiums for those with low incomes and requires insurers to accept all purchasers. This mixed private-public approach to universal coverage has emerged as a potential reform model for the United States. In this November 2007 interview, Dutch health minister Ab Klink discussed his country's system with Alain Enthoven. Enthoven is one of the chief architects of the "managed competition" model that laid the groundwork for the Clinton administration reform proposal in the 1990s and pending legislation in the Senate, as well as the current Dutch approach. PMID- 18397935 TI - Shepherding major health system reforms: a conversation with German health minister Ulla Schmidt. Interview by Tsung-Mei Cheng and Uwe Reinhardt. AB - Americans looking for role models among foreign health systems may find Germany's, along with the Dutch system, of particular interest. These countries seek to harvest the power of competition among nongovernmental insurers and providers of health care within a unifying regulatory framework aimed at keeping the competition fair and preserving for health care, as much as is possible, Europe's hallowed Principle of Solidarity. In this interview, conducted on behalf of Health Affairs by Tsung-Mei Cheng and Uwe Reinhardt, Germany's long-serving minister of health, Ulla Schmidt, explains what ethical and economic imperatives have shaped Germany's health reforms in recent years. PMID- 18397936 TI - Emerging role of cannabinoids in gastrointestinal and liver diseases: basic and clinical aspects. AB - A multitude of physiological effects and putative pathophysiological roles have been proposed for the endogenous cannabinoid system in the gastrointestinal tract, liver and pancreas. These range from effects on epithelial growth and regeneration, immune function, motor function, appetite control, fibrogenesis and secretion. Cannabinoids have the potential for therapeutic application in gut and liver diseases. Two exciting therapeutic applications in the area of reversing hepatic fibrosis as well as antineoplastic effects may have a significant impact in these diseases. This review critically appraises the experimental and clinical evidence supporting the clinical application of cannabinoid receptor-based drugs in gastrointestinal, liver and pancreatic diseases. Application of modern pharmacological principles will most probably expand the selective modulation of the cannabinoid system peripherally in humans. We anticipate that, in addition to the approval in several countries of the CB(1) antagonist, rimonabant, for the treatment of obesity and associated metabolic dysfunctions, other cannabinoid modulators are likely to have an impact on human disease in the future, including hepatic fibrosis and neoplasia. PMID- 18397937 TI - Management of Chronic Pain: Improving shared decision making in osteoarthritis. PMID- 18397938 TI - Multiple Health Problems in Elderly People: Adapting what is known. PMID- 18397939 TI - Excessive Drinking in Young Women: Reducing harm through quality improvement. PMID- 18397940 TI - Excessive Drinking in Young Women: Not just a "lifestyle disease". PMID- 18397941 TI - Palliative Care Beyond Cancer: Reliable comfort and meaningfulness. PMID- 18397942 TI - Palliative Care Beyond Cancer: Care for all at the end of life. PMID- 18397943 TI - Drug Resistant Infections in Poor Countries: A shrinking window of opportunity. PMID- 18397944 TI - Multiple Health Problems in Elderly People: Never had it so good? PMID- 18397945 TI - Management of Chronic Pain: Help and hope at the bottom of the pile. PMID- 18397946 TI - Adverse Drug Reactions in Elderly People: The challenge of safer prescribing. PMID- 18397947 TI - Drug Resistant Infections in Poor Countries: A major burden on children. PMID- 18397948 TI - Adverse Drug Reactions in Elderly People: A substantial cause of preventable illness. PMID- 18397949 TI - Making a difference: running the gauntlet to improve health care. PMID- 18397950 TI - End-bridging is required for pol mu to efficiently promote repair of noncomplementary ends by nonhomologous end joining. AB - DNA polymerase mu is a member of the mammalian pol X family and reduces deletion during chromosome break repair by nonhomologous end joining (NHEJ). This biological role is linked to pol mu's ability to promote NHEJ of ends with noncomplementary 3' overhangs, but questions remain regarding how it performs this role. We show here that synthesis by pol mu in this context is often rapid and, despite the absence of primer/template base-pairing, instructed by template. However, pol mu is both much less active and more prone to possible template independence in some contexts, including ends with overhangs longer than two nucleotides. Reduced activity on longer overhangs implies pol mu is less able to synthesize across longer gaps, arguing pol mu must bridge both sides of gaps between noncomplementary ends to be effective in NHEJ. Consistent with this argument, a pol mu mutant defective specifically on gapped substrates is also less active during NHEJ of noncomplementary ends both in vitro and in cells. Taken together, pol mu activity during NHEJ of noncomplementary ends can thus be primarily linked to pol mu's ability to work together with core NHEJ factors to bridge DNA ends and perform a template-dependent gap fill-in reaction. PMID- 18397951 TI - Exalign: a new method for comparative analysis of exon-intron gene structures. AB - The evolution of genes is usually studied and reconstructed at the sequence level, that is, by comparing and aligning their genomic, transcript or protein sequences. However, including the exon-intron structure of genes in the analysis can provide further and useful information, for example to draw reliable phylogenetic relationships left unsolved by traditional sequence-based evolutionary studies, or to shed further light on patterns of intron gain and loss. In spite of this, no tool especially devised for this task is currently available. In this work we present Exalign, an algorithm designed to retrieve, compare and search for the exon-intron structure of existing gene annotations, that has been implemented in a software tool freely accessible through a web interface as well as available for download. We present different applications of our method, from the reconstruction of the evolutionary history of homologous gene families to the detection of as of today unknown cases of intron loss in human and rodents, and, remarkably, two never reported intron gain events in human and mouse. The web interface for accessing Exalign is available at http://www.pesolelab.it/exalign/ or http://www.beacon.unimi.it/exalign/ PMID- 18397953 TI - Effectiveness of a brief intervention based on the '5A' model for smoking cessation at the primary care level in Santiago, Chile. AB - Chilean women have the highest smoking rates in Latin America. Prevalence in this population is about 40%. There are no national programs for smoking cessation at the primary care level. This study explores the feasibility and effectiveness of a brief counseling intervention targeted to women smokers of childbearing age who seek primary care in Santiago, Chile. A quasi-experimental design was used to compare the effect of an intervention based on the '5A' model developed by the National Cancer Institute in the United States and the standard care provided in two control clinics. Women smokers seeking care at the three primary care clinics were contacted during a 2 months period and offer to participate in the study. Sampling was stratified according to the age groups to ensure comparability between cohorts. Quotas were calculated for each age group. Participants were asked about their willingness to quit, self-efficacy, smoking behavior, addiction level as well as support received for smoking cessation. After 18 months of intervention all women were re-evaluated. A total of 773 women were recruited for the study; 76% of them completed the trial. Women smokers are characterized by a large percentage of light smokers with a low self-efficacy for quitting and with very low information on where and how to get assistance to quit. At study end, 15.2% of women reported quitting smoking at least for 1 month in the intervention clinic versus 7.8% in one of the control clinics (p < 0.05) and 14.6% in the second control clinic (p = NS). Over 70% of women in the intervention clinic were asked, assessed and received advice for quitting in comparison with <15% in the control clinics (p < 0.01). To conclude, a primary care intervention based on the '5A' model for smoking cessation is feasible and can have a significant effect in reducing smoking prevalence in this population. PMID- 18397952 TI - Lateral transfer of introns in the cryptophyte plastid genome. AB - Cryptophytes are unicellular eukaryotic algae that acquired photosynthesis secondarily through the uptake and retention of a red-algal endosymbiont. The plastid genome of the cryptophyte Rhodomonas salina CCMP1319 was recently sequenced and found to contain a genetic element similar to a group II intron. Here, we explore the distribution, structure and function of group II introns in the plastid genomes of distantly and closely related cryptophytes. The predicted secondary structures of six introns contained in three different genes were examined and found to be generally similar to group II introns but unusually large in size (including the largest known noncoding intron). Phylogenetic analysis suggests that the cryptophyte group II introns were acquired via lateral gene transfer (LGT) from a euglenid-like species. Unexpectedly, the six introns occupy five distinct genomic locations, suggesting multiple LGT events or recent transposition (or both). Combined with structural considerations, RT-PCR experiments suggest that the transferred introns are degenerate 'twintrons' (i.e. nested group II/group III introns) in which the internal intron has lost its splicing capability, resulting in an amalgamation with the outer intron. PMID- 18397954 TI - Sociodemographic and smoking associated with obesity in adult women in Iran: results from the National Health Survey. AB - BACKGROUND: There is no study that had a sample size sufficient to study the association between sociodemographic and smoking with obesity in Iran. The goal was to investigate these associations in the Iranian women. METHODS: Multivariate statistical techniques included 14 176 women between 20 and 69 years of age. Height and weight were measured rather than self-reported. RESULTS: In Iranian adult women, obesity OR(S) for the moderate and high education were 0.78 and 0.41, respectively, compared with basic level. Using low economy index as the reference, Obesity OR(S) for the urban women were 1.29, 1.25 and 1.28 for the lower-middle, upper-middle and high groups, respectively. Obesity OR(S) for the rural women were 1.71, 1.71 and 2.02 for the lower-middle, upper-middle and high groups, respectively. Obesity OR was 0.48 for active workforce compared with inactive group. Obesity OR was 0.70 for smokers women compared with nonsmokers. Using non-married as the reference group, Obesity OR(S) were 1.23 and 2.34 for married urban and rural women, respectively. CONCLUSIONS: Our results on the associations between age, smoking, education level, workforce and obesity are consistent with most studies, but between economic level and obesity are consistent with some study in developing countries. PMID- 18397955 TI - A dose-escalation study of rituximab for treatment of systemic lupus erythematosus and Evans' syndrome: immunological analysis of B cells, T cells and cytokines. AB - OBJECTIVE: Accumulating evidence suggests that B-cell depletion therapy by rituximab may be effective for autoimmune disorders. However, an optimal dose of rituximab and a mechanism of its action remain to be established. We performed a dose-escalation study for treatment of Japanese patients with autoimmune diseases including eight with SLE and one with Evans' syndrome. METHODS: Rituximab was infused intravenously, weekly 4 times in a dose-escalating fashion at three different doses of 100, 250 or 375 mg/m(2) to three patients each. Immunological parameters were monitored at certain points until 12 months after the treatment. RESULTS: Rituximab was well tolerated and safe in these patients. Seven out of eight SLE patients and one with Evans' syndrome clinically responded completely or partially to the treatment. Four patients achieved long-term remission (18-30 months) without any additional treatment. In these patients, a significant decrease in circulating B cells continued for 6 months after the treatment. The mean fluorescence intensities of CD19, CD21, CD40 and BR3 on the residual B cells as well as the percentage of CD69+ CD4+ T cells decreased significantly. Serum TNF-alpha levels decreased significantly on day 2. The Th1/Th2 balance of CD4+ T cells gradually shifted towards a Th1 type by 6 months. CONCLUSION: In addition to B-cell depletion, modification of B-cell and T-cell phenotypes as well as cytokine profiles may be involved in the action of rituximab. PMID- 18397956 TI - Expression of IL-19 and its receptors in RA: potential role for synovial hyperplasia formation. AB - OBJECTIVE: IL-19 is a novel cytokine of the IL-10 family. In this study, we sought to examine whether IL-19 plays a role in the pathogenesis of RA. METHODS: Expression of IL-19, IL-20 receptor 1 (IL-20R1) and IL-20R2 was examined by RT PCR and immunohistochemical analysis in rheumatoid synovium. The effects of IL-19 on synovial cells established from rheumatoid synovium (RASCs), with regard to IL 6 production and signal transducers and activators of transcription3 (STAT3) activation, were examined by ELISA and western blot analysis, respectively. The effect of IL-19 on RASC apoptosis was examined by Hoechst staining, flow cytometry analysis of annexin V binding and caspase-3 activity. RESULTS: IL-19, IL-20R1 and IL-20R2 mRNA were detected by RT-PCR in synovial tissues from RA patients. Immunohistochemical analysis showed IL-19 was predominantly expressed in the hyperplastic lining layers of RA synovial tissues. The majority of IL-19 positive cells were vimentin-positive and CD68-positive synovial cells, serving as markers of fibroblasts and macrophages, respectively. IL-20R1 and IL-20R2 (IL 20Rs) were expressed in both the lining and sublining layers of RA synovium. In RASC, IL-19 was induced by lipopolysaccharide stimulation and constitutive expression of IL-20Rs was observed, suggesting IL-19 has an autocrine action. In terms of this function, IL-19 induced STAT3 activation and increased IL-6 production by RASC above the medium control. Moreover, IL-19 significantly reduced RASC apoptosis induced by serum starvation. CONCLUSIONS: These data suggest that IL-19, produced by synovial cells, promotes joint inflammation in RA by inducing IL-6 production and decreasing synovial cell apoptosis. PMID- 18397957 TI - Investigation of the role of IL-1 and TNF in matrix degradation in the intervertebral disc. AB - OBJECTIVE: To establish if IL-1 or TNF regulates matrix degradation in the non degenerate or degenerate intervertebral disc (IVD). METHODS: In situ zymography (ISZ) has been used to investigate the role of IL-1 and TNF in the matrix degradation characterizing symptomatic IVD degeneration. ISZ employed three substrates (gelatin, collagen II, casein) and four different challenges, IL 1beta, IL-1 receptor antagonist (IL-1Ra), TNF-alpha and anti-TNF. RESULTS: We have shown for the first time that whilst IL-1beta will stimulate and IL-1 receptor antagonist will inhibit matrix degradation in intact human IVD tissue, neither TNF-alpha nor anti-TNF have any measurable effect on degradation of these matrices. CONCLUSION: This study has addressed a current area of controversy in IVD biology, namely, whether either IL-1 or TNF or both are involved in driving matrix degradation. Our data indicate that IL-1 is a key cytokine mediating matrix degradation in the IVD and therefore a therapeutic target. PMID- 18397958 TI - Eotaxin-3 is involved in Churg-Strauss syndrome--a serum marker closely correlating with disease activity. AB - OBJECTIVE: Churg-Strauss Syndrome (CSS) is characterized by excessive eosinophil accumulation in peripheral blood and affected tissues with development of granulomatous vasculitic organ damage. The contribution of eosinophil-chemotactic cytokines (eotaxin family) to eosinophilia and disease activity in CSS is unknown. Thus, we compared serum levels of the eotaxin family members in CSS patients with healthy and disease controls. METHODS: Forty patients with CSS diagnosed according to ACR 1990 criteria, 30 healthy controls (HC) and 57 disease controls (28 asthma, 20 small vessel vasculitis, 9 hypereosinophilic syndrome) were studied. Clinical data were collected and serum levels of eotaxin-1, -2 and 3 were determined by ELISA. Further, immunohistochemistry was applied to identify eotaxin-3 expression in tissue biopsies from patients with CSS. RESULTS: In contrast to eotaxin-1 and -2, eotaxin-3 was highly elevated in serum samples of active CSS patients and correlated highly significantly with eosinophil counts, total immunoglobulin E (IgE) levels and acute-phase parameters. Moreover, eotaxin 3 was not elevated in other eosinophilic and vasculitic diseases. Immunohistochemical analysis revealed strong expression of eotaxin-3 in endothelial and inflammatory cells in affected tissues of active CSS patients. CONCLUSIONS: This study reveals the specific association of elevated eotaxin-3 expression with high disease activity and eosinophilia in CSS patients. Eotaxin-3 might thus be a pathogenic player, biomarker and potential therapeutic target in CSS. PMID- 18397959 TI - Increased frequency of CD16+ monocytes and the presence of activated dendritic cells in salivary glands in primary Sjogren syndrome. AB - OBJECTIVES: In the salivary glands of patients with primary Sjogren Syndrome (pSjS) an accumulation of dendritic cells (DCs) is seen, which is thought to play a role in stimulating local inflammation. Aberrancies in subsets of monocytes, generally considered the blood precursors for DCs, may play a role in this accumulation of DCs. This study is aimed at determining the level of mature CD14lowCD16+ monocytes in pSjS and their contribution to the accumulation of DCs in pSjS. METHODS: Levels of mature and immature monocytes in patients with pSjS (n = 19) and controls (n = 15) were analysed by flow cytometry. The reverse transmigration system was used for generation of DCs generated from monocyte subsets. The phenotype of DCs in pSjS salivary glands was analysed using immunohistochemistry. In vivo tracking of monocyte subsets was performed in a mouse model. RESULTS: Increased levels of mature CD14lowCD16+ monocytes were found in patients with pSjS (mean (SD) 14.5 (5.5)% vs 11.4 (3.4)%). These cells showed normal expression of chemokine receptor and adhesion molecules. Mature monocytes partly developed into DC-lysosome-associated membrane glycoprotein (LAMP)+ (19.6 (7.5)%) and CD83+ (16 (9)%) DCs, markers also expressed by DCs in pSjS salivary glands. Monocyte tracking in the non-obese diabetic (NOD) mouse showed that the homologue population of mature mouse monocytes migrated to the salivary glands, and preferentially developed into CD11c+ DCs in vivo. CONCLUSIONS: Mature monocytes are increased in pSjS and patient and mouse data support a model where this mature monocyte subset migrates to the salivary glands and develops into DCs. PMID- 18397960 TI - Leflunomide and its metabolite A771726 are high affinity substrates of BCRP: implications for drug resistance. AB - BACKGROUND: Earlier publications have suggested a possible role for the efflux transporter breast cancer resistance protein (BCRP) in acquired resistance to disease-modifying antirheumatic drugs (DMARDs) such as leflunomide and its metabolite A771726 (teriflunomide). However, there is no direct evidence that BCRP interacts with these drugs. OBJECTIVES: To characterise the interaction between BCRP transporter and leflunomide and its active metabolite A771726, with emphasis on the nature of the interaction (substrate or inhibitor) and the kinetic characterisation of the interactions. METHODS: Different in vitro membrane-based methods (ATPase and vesicular transport assay) using BCRP-HAM-Sf9 membrane preparations and cellular assays (Hoechst assay and cytotoxicity assay) were performed on PLB985-BCRP and HEK293-BCRP cell lines overexpressing BCRP. RESULTS: In all assays used, an interaction between the investigated drugs and BCRP was detected. In the vesicular transport assay, both leflunomide and its metabolite inhibited BCRP-mediated methotrexate transport. Both compounds are likely substrates of BCRP as shown by the vanadate-sensitive ATPase assay. In line with the membrane assays, leflunomide and A771726 inhibited BCRP-mediated Hoechst efflux from PLB985-BCRP cells. In the cytotoxicity assay, overexpression of BCRP conferred 20.6-fold and 7.5-fold resistance to HEK293 cells against leflunomide and A771726, respectively. The resistance could be reversed by Ko134, a specific inhibitor of BCRP. CONCLUSION: Based on these results, BCRP could play an important role in the resistance to leflunomide and A771726 via interactions with these drugs. BCRP may also mediate drug-drug interactions when leflunomide is administered with other BCRP substrate drugs such as methotrexate. PMID- 18397961 TI - Involvement of a disintegrin and a metalloproteinase 8 (ADAM8) in osteoclastogenesis and pathological bone destruction. AB - OBJECTIVES: The eventual role of a disintegrin and a metalloproteinase 8 (ADAM8) in osteoclastogenesis was studied in erosive rheumatoid arthritis (RA) and in vitro. METHODS: ADAM8 protein and mRNA expression was measured in RA pannus and synovitis and compared to osteoarthritic (OA) synovial membrane. Human monocytes were isolated and stimulated with proinflammatory cytokines and their ADAM8 expression and surface ADAM8 were measured. Human peripheral blood monocytes and RAW 264.7 mouse monocyte/macrophage cells were stimulated to osteclast like cells, and their expression of ADAM8 and osteoclastic markers (calcitonin receptor, integrin beta 3, cathepsin K, TRAP) were analysed. Transfection and small interfering RNA (siRNA) were used to assess the role of ADAM8 in formation of polykaryons. RESULTS: Increased numbers of ADAM8 positive cells were shown particularly in the pannus-cartilage/bone junction close or adjoining to TRAP positive multinucleate cells under formation (60 (2)% in pannus, 47 (2)% in synovitis vs 10 (1)% in OA, p<0.001). Human pannus contained high ADAM8 mRNA copy numbers (23 (7) in pannus, 14 (4) in synovitis vs 1.7 (0.3) in OA, p<0.001). Functional studies in vitro disclosed ADAM8 mRNA and protein, which was first converted to a proteolytically active and then to fusion-active form. Gene transfection and siRNA experiments enhanced and inhibited, respectively, expression of osteoclast markers and maturation of multinuclear cells. CONCLUSIONS: ADAM8 may be involved in bone destruction in RA because it is upregulated in RA pannus adjacent to developing erosions and enhances maturation of osteoclast-like cells. PMID- 18397963 TI - Coupling erythropoietin secretion to mesenchymal stromal cells enhances their regenerative properties. AB - AIMS: Mesenchymal stromal cells (MSCs) possess intrinsic features that identify them as useful for treating ischaemic syndromes. Poor in vivo survival/engraftment of MSCs, however, limits their overall effectiveness. In this work, we tested whether genetically engineering MSCs to secrete erythropoietin (Epo) could represent a better therapeutic platform than MSCs in their native form. METHODS AND RESULTS: MSCs from C57Bl/6 mice were retrovirally transduced with either an empty vector or one that causes the production of Epo and were then analysed for the alterations in angiogenic and survival potential. Using a mouse model of myocardial infarction (MI), the regenerative potential of null MSCs and Epo-overexpressing MSCs (Epo+MSCs) was assessed using serial echocardiogram and invasive haemodynamic measurements. Infarct size, capillary density and neutrophil influx were assessed using histologic techniques. Using in vitro assays coupled with an in vivo Matrigel plug assay, we demonstrate that engineering MSCs to express Epo does not alter their immunophenotype or plasticity. However, relative to mock-modified MSCs [wild-type (WT)-MSCs], Epo+MSCs are more resilient to apoptotic stimuli and initiate a more robust host derived angiogenic response. We also identify and characterize the autocrine loop established on MSCs by having them secrete Epo. Furthermore, in a murine model of MI, animals receiving intracardiac injections of Epo+MSCs exhibited significantly enhanced cardiac function compared with WT-MSCs and saline-injected control animals post-MI, owing to the increased myocardial capillary density and the reduced neutrophilia. CONCLUSION: Epo overexpression enhances the cellular regenerative properties of MSCs by both autocrine and paracrine pathways. PMID- 18397964 TI - Pulmonary hyperplasia and the two sides of PKCzeta. PMID- 18397962 TI - Overexpression of heat shock protein 27 protects against ischaemia/reperfusion induced cardiac dysfunction via stabilization of troponin I and T. AB - AIMS: Heat shock protein 27 (Hsp27) renders cardioprotection from ischaemia/reperfusion (I/R) injury, but little is known about its role in myofilaments. We proposed that increased expression of Hsp27 may improve post ischaemic contractile dysfunction by preventing I/R-induced cardiac troponin I (cTnI) and troponin T (cTnT) degradation. METHODS AND RESULTS: Adenovirus mediated Hsp27 overexpression improved contractile function in perfused rat hearts subjected to global no-flow I/R (30-min/30-min). Such improvement was further confirmed in Hsp27-overexpressing cardiomyocytes subjected to simulated I/R (20-min/30-min). Moreover, these cells showed restored myofilament response to Ca(2+) but not intracellular Ca(2+) transients. The protection correlated with attenuation of I/R-induced cTnI and cTnT degradation. Confocal microscopy revealed co-localization of Hsp27 with these proteins. Co-immunoprecipitation and pull-down assays further confirmed that Hsp27 interacted with the COOH-terminus of cTnI and the NH(2)-terminus of cTnT and that Hsp27 overexpression decreased the interaction between mu-calpain (a protease mediating proteolysis of cTnI and cTnT) and cTnI or cTnT under I/R. CONCLUSION: The findings reveal a novel role of Hsp27 in the protection of cTnI and cTnT from I/R-induced degradation by preventing their proteolytic cleavage via interacting with these proteins. Such protection may result in restored post-ischaemic myofilament response to Ca(2+) and improved post-ischaemic contractile function. PMID- 18397965 TI - Benefits and dangers of university-sponsored open-access systems. PMID- 18397966 TI - Noninvasive imaging of quantitative cerebral blood flow changes during 100% oxygen inhalation using arterial spin-labeling MR imaging. AB - BACKGROUND AND PURPOSE: Tracer studies have demonstrated that 100% oxygen inhalation causes a small cerebral blood flow (CBF) decrease. This study was performed to determine whether arterial spin-labeling (ASL), a noninvasive MR imaging technique, could image these changes with clinically reasonable imaging durations. MATERIALS AND METHODS: Continuous ASL imaging was performed in 7 healthy subjects before, during, and after 100% oxygen inhalation. ASL difference signal intensity (DeltaM, control - label), CBF, and CBF percentage change were measured. A test-retest paradigm was used to calculate the variability of the initial and final room air CBF measurements. RESULTS: During oxygen inhalation, DeltaM decreased significantly in all regions (eg, global DeltaM decreased by 23 +/- 11%, P < .01, all values mean +/- SD). Accounting for the reduced T1 of hyperoxygenated blood, we found a smaller CBF decrease, which did not reach significance in any of the regions. Global CBF dropped from 50 +/- 10 mL per 100 g/minute to 47 +/- 10 mL per 100 g/minute following 100% oxygen inhalation, a decrease of 5 +/- 14% (P > .17). The root-mean-square variability of the initial and final room air CBF measurements was 7-8 mL per 100 g/minute. CONCLUSIONS: The DeltaM signal intensity decreased significantly with oxygen inhalation; however, after accounting for changes in blood T1 with oxygen, CBF decreases were small. Such measurements support the use of hyperoxia as an MR imaging contrast agent and may be helpful to interpret hyperoxia-based stroke trials. PMID- 18397967 TI - Arteriovenous shunt visualization in arteriovenous malformations with arterial spin-labeling MR imaging. AB - BACKGROUND AND PURPOSE: A reliable quantitative technique for measuring arteriovenous (AV) shunt in vascular malformations is not currently available. Here, we evaluated the hypothesis that continuous arterial spin-labeled (CASL) perfusion MR imaging can be used to detect and measure AV shunt in patients with arteriovenous malformations (AVMs). MATERIALS AND METHODS: CASL perfusion MR imaging was performed in 7 patients with AVMs. Semiquantitative AV shunt estimates were generated based on a thresholding strategy by using signal intensity difference (DeltaM) images to avoid potential errors in cerebral blood flow (CBF) calculation related to abnormal transit times and nonphysiologic blood tissue water exchange in and around the AVMs. The potential for measuring CBF in regions distant from and near the AVM was explored, as was the relationship of CBF changes related to the size of the shunt. RESULTS: In all 7 cases, striking increased intensity was seen on CASL perfusion DeltaM maps in the nidus and venous structures draining the AVM. Shunt estimates ranged from 30% to 0.6%. Mean CBF measurements in structures near the AVMs were not significantly different from the contralateral measurements. However, CBF in adjacent ipsilateral white matter increased relative to the contralateral side as the percent shunt increased (P = .02). Cortical gray matter CBF Delta (contralateral-ipsilateral) values demonstrated the same effect, but the correlation was weak and not significant. Thalamic CBF decreased ipsilaterally with increasing percent AV shunt (P = .01), indicating a possible steal effect. Basal ganglia Delta values showed little change in CBF with the size of the AV shunt. CONCLUSION: CASL perfusion MR imaging can demonstrate AV shunting, providing high lesion conspicuity and a novel means for evaluating AVM physiology. PMID- 18397968 TI - Low-dose nonenhanced head CT protocol for follow-up evaluation of children with ventriculoperitoneal shunt: reduction of radiation and effect on image quality. AB - BACKGROUND AND PURPOSE: Children with a shunt for hydrocephalus often undergo multiple follow-up head CT scans, increasing the risk for long-term effects of ionizing radiation. The purpose of our study was to evaluate if an unenhanced low dose head CT could consistently provide acceptable image quality and diagnostic information. MATERIALS AND METHODS: Ninety-two children (mean age, 9 years; range, 8 months to 21 years; 45 boys and 47 girls) with a shunt for hydrocephalus and no clinical evidence of shunt malfunction who were referred for a follow-up nonenhanced head CT were included in the study. All studies were performed on a 4 section multidetector CT. Two CT studies were selected retrospectively for each patient, 1 performed at standard dose (220 mA) and 1 at low dose (80 mAs). Two radiologists independently evaluated and graded both standard-dose and low-dose studies for various image quality parameters. Attenuation and noise levels were measured, and gray-white differentiation and contrast-to-noise ratio (CNR) were calculated. RESULTS: Low-dose CT resulted in 63% mean dose reduction. All low dose CT scans were diagnostically acceptable. Image quality parameters were significantly lower at low dose (P = .0001) except for the parameters for streak artifacts (P = .46) and need for further imaging (P = .47), which were higher. Mean noise levels were significantly higher (P = .001) in low-dose studies, whereas CNR was significantly higher in standard dose CT (P = .001). A moderate to perfect agreement was noted between the 2 readers with regard to image quality assessment (65%-99%). CONCLUSION: Low-dose nonenhanced head CT consistently provides diagnostically acceptable images with relevant diagnostic information in children with VP shunts resulting in substantial dose savings. PMID- 18397969 TI - Clinical and ultrasonographic correlation between scapular dyskinesia and subacromial space measurement among junior elite tennis players. AB - OBJECTIVES: In this study, the hypothesis that tennis players with scapular dyskinesia present a smaller subacromial space than non-athletes was investigated. Additionally, the correlation between the size of the subacromial space and abnormalities in scapular movement during arm abduction was studied. DESIGN: Cross-sectional study. PARTICIPANTS: A total of 53 elite tennis players and 20 control participants were enrolled in the study. Participation was restricted to elite-level, junior tennis players who had no current shoulder pain or history of shoulder injuries. INTERVENTION: Each individual was examined for scapular dyskinesia by a single physician and by ultrasound, with the results analysed in a blind fashion by a single radiologist. RESULTS: 43.4% of the tennis players and 20% of control participants presented with scapular dyskinesia. Of the 106 shoulders evaluated, 39.6% of tennis players and 10% of control participants presented with scapular dyskinesia in the clinical examination (p = 0.005). Ultrasonographic measurements demonstrated that tennis players presented statistically smaller subacromial spaces compared with control participants (p<0.001). A decrease in the subacromial space was observed in tennis players when the shoulder was raised from 0 degrees to 60 degrees of abduction; however, dyskinesia-afflicted athletes demonstrated a significantly greater decrease following this movement (19.3 vs 13.8 mm, p = 0.002). CONCLUSIONS: The results of this study demonstrated that tennis players with scapular dyskinesia present a smaller subacromial space than control participants. Furthermore, when the shoulder was analysed dynamically, moving from neutral abduction to 60 degrees of elevation, the tennis players with scapular dyskinesia presented a greater reduction in the subacromial space compared with unaffected athletes. PMID- 18397970 TI - Gait-related intrinsic risk factors for patellofemoral pain in novice recreational runners. AB - OBJECTIVE: To determine prospectively gait-related intrinsic risk factors for patellofemoral pain (PFP) in a population of novice recreational runners. DESIGN: Prospective cohort study. PARTICIPANTS: 102 novice recreational runners (89 women) with no history of knee or lower leg complaints. INTERVENTIONS: The standing foot posture of the subjects was examined and plantar pressure measurements during running were collected. The subjects then participated in a 10-week "start to run" programme. During this period all sports injuries were registered by a sports medicine physician. MAIN OUTCOME MEASUREMENTS: The relationship between the standing foot posture and PFP was investigated and gait related intrinsic risk factors for PFP were determined. RESULTS: The 17 runners who developed PFP exerted a significantly higher vertical peak force underneath the lateral heel and metatarsals 2 and 3. Logistic regression analysis showed that a significantly higher vertical peak force underneath the second metatarsal and shorter time to the vertical peak force underneath the lateral heel were predictors for PFP. No significant evidence was found for an association between an excessively pronated or supinated foot posture and the development of PFP. CONCLUSIONS: The findings suggest that an excessive impact shock during heel strike and at the propulsion phase of running may contribute to an increased risk of developing PFP. The hypothesis that persons at risk for PFP show an altered static foot posture in comparison with non-afflicted persons is not supported by the results of this study. PMID- 18397971 TI - Patients' perceptions of electronic monitoring devices affect medication adherence in hypertensive African Americans. AB - BACKGROUND: Electronic monitoring devices (EMDs) are regarded as the gold standard for assessing medication adherence in clinical research. However, little is known about the effect of patients' acceptance of EMDs on medication adherence in African Americans with hypertension who are followed in primary care practices OBJECTIVE: To assess patients' perceptions of EMDs, their acceptance of EMDs, and the relationship of these perceptions to medication adherence in African Americans with hypertension who are followed in community-based practices. METHODS: Patients were recruited from a larger randomized controlled trial assessing the effect of motivational interviewing on medication adherence and blood pressure in hypertensive African American patients followed in 2 New York City primary care practices. Medication adherence was assessed with a Medication Event Monitoring System (MEMS) during a 12-month monitoring period. At the 12 month follow-up, patients' perceptions of the MEMS were assessed with a 17-item questionnaire. ANOVA was used to compare patients' responses (agree, neither, disagree) with the MEMS adherence over the monitoring period. Tukey's post hoc tests were used to determine whether there were significant differences among the 3 groups. RESULTS: Participants were predominantly women, low-income, unemployed, had a high school education, and were a mean age of 53 years. Approximately two thirds of the participants stated that the MEMS helped them remember to take their medications, 93% reported that the MEMS was easy to open, 85% did not find it stressful, and 75% liked the MEMS and used it everyday. One-third of patients preferred using a pillbox and 25% did not like traveling with the MEMS. Patients who stated that they used the MEMS every day, felt comfortable using it in front of others, and remembered to put refills in the MEMS had significantly better adherence over the study period than did those who disagreed (p 20 mg/dL). The patient died 3 months after admission. DISCUSSION: Rosiglitazone maleate is a thiazolidinedione approved for treatment of type 2 diabetes mellitus. The first member of this drug class, troglitazone, was withdrawn from the market due to reports of acute liver failure. Rosiglitazone has been shown to be much safer than troglitazone, despite some reported cases of early-onset nonfatal hepatotoxicity. Use of the Naranjo probability scale indicated that rosiglitazone was the probable cause of acute liver failure in our patient. CONCLUSIONS: We conclude that rosiglitazone may be associated with late-onset acute liver failure. Clinicians should be aware of such a complication and monitor liver function in patients receiving the drug. PMID- 18397975 TI - The common P450 oxidoreductase variant A503V is not a modifier gene for 21 hydroxylase deficiency. AB - CONTEXT: 21-hydroxylase deficiency (21OHD) is a common genetic disorder caused by mutations in the CYP21A2 gene, which encodes the adrenal 21-hydroxylase, microsomal P450c21. CYP21A2 gene mutations generally correlate well with impaired P450c21 enzymatic activity and the clinical findings in 21OHD, but occasional discrepancies between genotype and phenotype suggest the effects of modifier genes. Mutations in P450 oxidoreductase (POR), the protein that transfers electrons from reduced nicotinamide adenine dinucleotide phosphate to all microsomal P450s, can ameliorate the 21OHD phenotype and, therefore, could be a modifier gene. OBJECTIVES: We sought to identify POR variants in patients with 21OHD having discordant phenotype and genotype, and to evaluate their effect on 21-hydroxylase activity. PATIENTS AND METHODS: We determined the CYP21A2 genotypes of 313 Brazilian patients with 21OHD and correlated the genotype and phenotype. The POR gene was sequenced in 17 patients with discordant genotype and phenotype. Wild-type and A503V POR, and P450c21 were expressed in bacteria and reconstituted in vitro. Activities were assayed by conversion of [(14)C]progesterone to deoxycorticosterone and [(3)H]17-hydroxyprogesterone to 11 deoxycortisol, and assessed by thin layer chromatography and phosphorimaging. RESULTS: The A503V POR variant was found in 10 of 30 alleles, the same ratio as in the normal population. There were no significant differences in Michaelis constant, maximum velocity and maximum velocity/Michaelis constant of 21 hydroxylase activity supported by wild-type and A503V POR. CONCLUSION: The only POR missense polymorphism found in atypical 21OHD patients was A503V. Although A503V reduces P450c17 enzymatic activity, it does not influence P450c21 activity, indicating that POR A503V does not modify the 21OHD phenotype. PMID- 18397976 TI - Muscle adaptation to short-term fasting in healthy lean humans. AB - CONTEXT: It has been demonstrated repeatedly that short-term fasting induces insulin resistance, although the exact mechanism in humans is unknown to date. Intramyocellular sphingolipids (i.e. ceramide) have been suggested to induce insulin resistance by interfering with the insulin signaling cascade in obesity. OBJECTIVE: Our objective was to study peripheral insulin sensitivity together with muscle ceramide concentrations and protein kinase B/AKT phosphorylation after short-term fasting. MAIN OUTCOME MEASURES AND DESIGN: After 14- and 62-h fasting, glucose fluxes were measured before and after a hyperinsulinemic euglycemic clamp. Muscle biopsies were performed in the basal state and during the clamp to assess muscle ceramide and protein kinase B/AKT. RESULTS: Insulin mediated peripheral glucose uptake was significantly lower after 62-h fasting compared with 14-h fasting. Intramuscular ceramide concentrations tended to increase during fasting. During the clamp the phosphorylation of protein kinase B/AKT at serine(473) in proportion to the total amount of protein kinase B/AKT was significantly lower. Muscle ceramide did not correlate with plasma free fatty acids. CONCLUSIONS: Fasting for 62 h decreases insulin-mediated peripheral glucose uptake with lower phosphorylation of AKT at serine(473). AKT may play a regulatory role in fasting-induced insulin resistance. Whether the decrease in AKT can be attributed to the trend to higher muscle ceramide remains unanswered. PMID- 18397977 TI - Autonomic nervous system function in chronic exogenous subclinical thyrotoxicosis and the effect of restoring euthyroidism. AB - CONTEXT: Knowledge on the relationship between the autonomic nervous system and subclinical hyperthyroidism is mainly based upon cross-sectional studies in heterogeneous patient populations, and the effect of restoration to euthyroidism in subclinical hyperthyroidism has not been studied. OBJECTIVE: We investigated the long-term effects of exogenous subclinical hyperthyroidism on the autonomic nervous system and the potential effects of restoration of euthyroidism. DESIGN: This was a prospective single-blinded, placebo-controlled, randomized trial. SETTING: The study was performed at a university hospital. PATIENTS: A total of 25 patients who were on more than 10-yr TSH suppressive therapy after thyroidectomy was examined. INTERVENTION: Patients were studied at baseline and subsequently randomized to a 6-month thyroid hormone substitution regimen to obtain either euthyroidism or maintenance of the subclinical hyperthyroid state. MAIN OUTCOME MEASURES: Urinary excretion of catecholamines and heart rate variability were measured. Baseline data of the subclinical hyperthyroidism patients were compared with data obtained in patients with hyperthyroidism and controls. RESULTS: Urinary excretion of norepinephrine and vanillylmandelic acid was higher in the subclinical hyperthyroidism patients compared with controls and lower compared with patients with overt hyperthyroidism. Heart rate variability was lower in patients with hyperthyroidism, intermediate in subclinical hyperthyroidism patients, and highest in the healthy controls. No differences were observed after restoration of euthyroidism. CONCLUSIONS: Long-term exogenous subclinical hyperthyroidism has effects on the autonomic nervous system measured by heart rate variability and urinary catecholamine excretion. No differences were observed after restoration to euthyroidism. This may indicate the occurrence of irreversible changes or adaptation during long-term exposure to excess thyroid hormone that is not remedied by 6-month euthyroidism. PMID- 18397978 TI - [123 I]Iodometomidate for molecular imaging of adrenocortical cytochrome P450 family 11B enzymes. AB - BACKGROUND: Due to advances in conventional imaging, adrenal tumors are detected with increasing frequency. However, conventional imaging provides only limited information on the origin of these lesions, which represent a wide range of different pathological entities. New specific imaging methods would therefore be of great clinical value. We, therefore, studied the potential of iodometomidate (IMTO) as tracer for molecular imaging of cytochrome P450 family 11B (Cyp11B) enzymes. METHODS: Inhibition of Cyp11B1 and Cyp11B2 by IMTO, etomidate, metomidate, and fluoroetomidate was investigated in NCI-h295 cells and in Y1 cells stably expressing hsCyp11B1 or hsCyp11B2. Pharmacokinetics and biodistribution after iv injection of [(123/125)I]IMTO were analyzed in mice in biodistribution experiments and by small-animal single-photon emission computed tomography (SPECT). Furthermore, four patients with known adrenal tumors (two metastatic adrenal adenocarcinomas, one bilateral adrenocortical adenoma, and one melanoma metastasis) were investigated with [(123)I]iodometomidate-SPECT. RESULTS: In cell culture experiments, all compounds potently inhibited both Cyp11B1 and Cyp11B2. Adrenals showed high and specific uptake of [(123/125)I]IMTO and were excellently visualized in mice. In patients, adrenocortical tissue showed high and specific tracer uptake in both primary tumor and metastases with short investigation time and low radiation exposure, whereas the non adrenocortical tumor did not exhibit any tracer uptake. CONCLUSION: We have successfully completed the development of an in vivo detection system of adrenal Cyp11B enzymes by [(123)I]IMTO scintigraphy in both experimental animals and humans. Our findings suggest that [(123)I]IMTO is a highly specific radiotracer for imaging of adrenocortical tissue. Due to the general availability of SPECT technology, we anticipate that [(123)I]IMTO scintigraphy may become a widely used tool to characterize adrenal lesions. PMID- 18397979 TI - Retinol-binding protein 4 and its relation to insulin resistance in obese children before and after weight loss. AB - CONTEXT: There are limited and controversial data concerning the relationships between retinol-binding protein 4 (RBP4), weight status, and insulin resistance in obese humans and especially in children. OBJECTIVE: Our objective was to study the longitudinal relationships among RBP4, insulin resistance and weight status in obese children. DESIGN, SETTING, AND PATIENTS: We conducted a 1-yr longitudinal follow-up study in a primary-care setting with 43 obese children (median age 10.8 yr) and 19 lean children of same the age and gender. INTERVENTION: Our outpatient 1-yr intervention program was based on exercise, behavior, and nutrition therapy. MAIN OUTCOMES MEASURES: Changes of weight status (body mass index sd score), RBP4, molar RBP4/serum retinol (SR) ratio, insulin resistance index homeostasis model assessment (HOMA), and quantitative insulin sensitivity check index (QUICKI). RESULTS: Obese children had significantly (P < 0.01) higher RBP4 concentrations and a higher RBP4/SR ratio compared with lean children. In multiple linear regression analyses adjusted to age, gender, and pubertal stage, RBP4 was significantly correlated to insulin and body mass index. Pubertal children demonstrated significantly decreased QUICKI and significantly increased HOMA index, insulin, and RBP4 concentrations compared with prepubertal children. Changes of RBP4 correlated significantly to changes of insulin (r = 0.29), HOMA index (r = 0.29), QUICKI (r = 0.22), and weight status (r = 0.31). Substantial weight loss in 25 children led to a significant (P < 0.001) decrease of RBP4, RBP4/SR, blood pressure, triglycerides, insulin, and HOMA index and an increase in QUICKI in contrast to the 18 children without substantial weight loss. CONCLUSION: RBP4 levels were related to weight status and insulin resistance in both cross-sectional and longitudinal analyses, suggesting a relationship between RBP4, obesity, and insulin resistance in children. PMID- 18397980 TI - Influence of the d3-growth hormone (GH) receptor isoform on short-term and long term treatment response to GH replacement in GH-deficient adults. AB - OBJECTIVE: Recombinant human GH (rhGH) replacement in adults is aimed at improving signs and symptoms of the adult GH deficiency (GHD) syndrome. In children, a common polymorphism of the GH receptor (exon-3 deletion, d3GHR) increases the response to rhGH replacement. The aim of this study was to assess the effects of this polymorphism on the response to rhGH replacement in adults. DESIGN: Prospective intervention with rhGH during 1 yr (n = 99) and in a subset during 5 yr (n = 53). PATIENTS AND METHODS: The presence of the d3GHR variant was established in GHD patients and linked to short-term and long-term effects of rhGH replacement on IGF-I, lipid metabolism, anthropometric parameters, and bone mineral density. RESULTS: Fifty-five patients had two wild-type alleles (56%), whereas 38 patients (38%) had one allele and six patients (6%) had two alleles coding the d3GHR isoform. During short-term rhGH replacement, the increase in IGF I was higher in patients bearing at least one d3GHR allele, compared with those with two wild-type alleles (at an identical mean dose of rhGH). The decrease in total cholesterol and low-density lipoprotein cholesterol was lower in the group bearing at least one d3GHR allele, whereas the increase in high-density lipoprotein cholesterol was higher, compared with patients with the wild-type genotype. In contrast, these differential responses of GHR genotype could not be demonstrated during long-term rhGH replacement. CONCLUSION: The d3GHR genotype contributes, at least for some parameters, to the interindividual differences in efficacy of short-term, but not long-term, rhGH replacement in adults with GHD. PMID- 18397981 TI - Baseline characteristics and the effects of two years of growth hormone (GH) replacement therapy in adults with GH deficiency previously treated for acromegaly. AB - CONTEXT: The effects of GH replacement in GH-deficient (GHD) adults previously treated for acromegaly are not well known. OBJECTIVE, DESIGN, AND PATIENTS: In this single-center, open-labeled, prospective study, 10 consecutive GHD adults with cured acromegaly (A group) and 10 matched GHD adults with previous nonfunctioning hypopituitary disease (NF group) were included. Comparisons were made at baseline and in the responses in body composition, muscle strength, bone mass, and metabolic indices during 2 yr of GH replacement. RESULTS: At baseline, upper leg local muscle endurance and serum low-density lipoprotein-cholesterol concentration were more impaired in the A group. The A group contained three patients with hypertension, one with diabetes mellitus type 2, and one with hyperlipidemia. The NF group had only one patient with hypertension. There were no significant between-group differences in the responses to the GH therapy. Body composition and serum lipid pattern improved in both groups without any deterioration of glucose homeostasis. At study end, no difference remained between the two groups in any variable. During the 2-yr treatment, one patient had a myocardial infarction and two had cerebral infarctions in the A group, whereas no vascular event occurred in the NF group. CONCLUSIONS: GHD patients with previous acromegaly have an impaired cardiovascular risk profile and decreased local muscle endurance as compared with other GHD patients. Two-year GH replacement eliminated these differences, but vascular events occurred more frequently in the A group. Therefore, GHD patients with cured acromegaly will benefit from GH replacement, but careful monitoring of cardiovascular status is needed. PMID- 18397982 TI - Adiponectin and left ventricular structure and function in healthy adults. AB - CONTEXT: Adiponectin inhibits protein synthesis in cardiac myocytes, thereby opposing the effect of cardiac workload and trophic factors (in particular, insulin) on left ventricular (LV) mass and wall thickness (WT). OBJECTIVE: We tested whether adiponectin and its isoforms are related to LV mass, WT, and function independently of metabolic factors. DESIGN: This was a cross-sectional study. SUBJECTS: The study included 77 healthy volunteers (42 men) aged 30-59 yr with normal LV structure and function. MAIN OUTCOME MEASURES: Insulin response and insulin sensitivity were assessed by oral glucose tolerance test and euglycemic hyperinsulinemic clamp. LV mass, WT, stroke work, chamber function, and myocardial longitudinal function were evaluated by standard Doppler echocardiography and tissue Doppler imaging. Total and molecular isoforms of adiponectin were measured in plasma. RESULTS: By multivariate analysis, independent factors affecting LV mass were sex, body mass index, stroke work, and current smoking (R(2) = 0.66). Independent correlates of LV WT were age, stroke work, and plasma adiponectin (standardized r = 0.28, 0.41, and -0.26, P at least < 0.005, R(2) = 0.48). LV longitudinal late diastolic velocity was independently related to age, body mass index, and adiponectin (standardized r = 0.20, 0.26, 0.33, P at least < 0.05, R(2) = 0.30). High-molecular-weight adiponectin (47% of total), but not lower molecular-weight isoforms, insulin sensitivity, or other metabolic factors, was inversely and independently related to WT (standardized r = -0.27, P < 0.01) and myocardial longitudinal late diastolic velocity (standardized r = -0.28, P < 0.05). CONCLUSION: In healthy subjects, circulating total and high-molecular-weight adiponectin are related to LV WT and diastolic function, independently of age and metabolic factors. PMID- 18397983 TI - Normal values of circulating insulin-like growth factor-I bioactivity in the healthy population: comparison with five widely used IGF-I immunoassays. AB - BACKGROUND: IGF-I immunoassays are primarily used to estimate IGF-I bioactivity. Recently an IGF-I-specific kinase receptor activation assay (KIRA) has been developed as an alternative method. However, no normative values have been established for the IGF-I KIRA. OBJECTIVE: The objective of the study was to establish normative values for the IGF-I KIRA in healthy adults. DESIGN: This was a cross-sectional study in healthy nonfasting blood donors. STUDY PARTICIPANTS: Participants included 426 healthy individuals (310 males, 116 females; age range 18-79 yr). MAIN OUTCOME MEASURES: IGF-I bioactivity determined by the KIRA was measured. Results were compared with total IGF-I, measured by five different IGF I immunoassays. RESULTS: Mean (+/- sd) IGF-I bioactivity was 423 (+/- 131) pmol/liter and decreased with age (beta = -3.4 pmol/liter.yr, P < 0.001). In subjects younger than 55 yr, mean IGF-I bioactivity was significantly higher in women than men. Above this age this relationship was inverse, suggesting a drop in IGF-I bioactivity after menopause. This drop was not reflected in total IGF-I levels. IGF-I bioactivity was significantly related to total IGF-I (r(s) varied between 0.46 and 0.52; P < 0.001). CONCLUSIONS: We established age-specific normative values for the IGF-I KIRA. We observed a significant drop in IGF-I bioactivity in women between 50 and 60 yr, which was not perceived by IGF-I immunoassays. The IGF-I KIRA, when compared with IGF-I immunoassays, theoretically has the advantage that it measures net effects of IGF-binding proteins on IGF-I receptor activation. However, it has to be proven whether information obtained by the IGF-I KIRA is clinically more relevant than measurements obtained by IGF-I immunoassays. PMID- 18397984 TI - Treatment of type 2 diabetes and dyslipidemia with the natural plant alkaloid berberine. AB - CONTEXT: Berberine, a natural plant alkaloid, is usually used as an antibiotic drug. The potential glucose-lowering effect of berberine was noted when it was used for diarrhea in diabetic patients. In vitro and in vivo studies have then showed its effects on hyperglycemia and dyslipidemia. OBJECTIVE: The objective of the study was to evaluate the efficacy and safety of berberine in the treatment of type 2 diabetic patients with dyslipidemia. DESIGN: One hundred sixteen patients with type 2 diabetes and dyslipidemia were randomly allocated to receive berberine (1.0 g daily) and the placebo for 3 months. The primary outcomes were changes in plasma glucose and serum lipid concentrations. Glucose disposal rate (GDR) was measured using a hyperinsulinemic euglycemic clamp to assess insulin sensitivity. RESULTS: In the berberine group, fasting and postload plasma glucose decreased from 7.0 +/- 0.8 to 5.6 +/- 0.9 and from 12.0 +/- 2.7 to 8.9 +/- 2.8 mm/liter, HbA1c from 7.5 +/- 1.0% to 6.6 +/- 0.7%, triglyceride from 2.51 +/- 2.04 to 1.61 +/- 1.10 mm/liter, total cholesterol from 5.31 +/- 0.98 to 4.35 +/- 0.96 mm/liter, and low-density lipoprotein-cholesterol from 3.23 +/- 0.81 to 2.55 +/- 0.77 mm/liter, with all parameters differing from placebo significantly (P < 0.0001, P < 0.0001, P < 0.0001, P = 0.001, P < 0.0001, and P <0.0001, respectively). The glucose disposal rate was increased after berberine treatment (P = 0.037), although no significant change was found between berberine and placebo groups (P = 0.063). Mild to moderate constipation was observed in five participants in the berberine group. CONCLUSIONS: Berberine is effective and safe in the treatment of type 2 diabetes and dyslipidemia. PMID- 18397985 TI - 3,5,3'-Triiodothyronine thyrotoxicosis due to increased conversion of administered levothyroxine in patients with massive metastatic follicular thyroid carcinoma. AB - OBJECTIVE: Some patients with massive metastatic thyroid carcinoma exhibit T(3) thyrotoxicosis. We investigated the prevalence and cause of T(3) thyrotoxicosis and the clues to the diagnosis. DESIGN: Serum free T(3) (FT(3)), free T(4) (FT(4)), and TSH were measured in patients with massive metastases from papillary, follicular, or medullary thyroid carcinomas (31, 20, and seven patients, respectively). Patients without recurrence served as controls. Thyrotoxic patients were reexamined 1 wk after withdrawal of levothyroxine. Type 1 and type 2 iodothyronine deiodinase (D1 and D2) activities were measured in three tumor tissues from thyrotoxic patients. MAIN OUTCOME: The serum FT(3) level and FT(3)/FT(4) ratio in the follicular carcinoma (FC) group were significantly higher than those in the papillary carcinoma group or patients without recurrence. Four patients (20%) in the FC group but none in the other groups demonstrated T(3) thyrotoxicosis or a FT(3)/FT(4) ratio greater than 3.5. One week after withdrawal of levothyroxine, both FT(3) and FT(4) levels decreased. Retrospective measurements of FT(3) in frozen stored sera demonstrated that FT(3) exceeded the upper normal limit when FT(4) began to decrease but remained within the normal range. Tumor tissues showed high D1 and D2 activities. CONCLUSIONS: Twenty percent of patients with massive metastatic FC exhibited T(3) thyrotoxicosis, most likely due to increased conversion of T(4) to T(3) by tumor expressing high D1 and D2 activities. Occasional measurement of serum FT(3) in addition to FT(4) and TSH is recommended in patients with massive metastatic FC, especially when serum FT(4) decreases on fixed doses of levothyroxine. PMID- 18397986 TI - Insulin intervention in slowly progressive insulin-dependent (type 1) diabetes mellitus. AB - OBJECTIVE: We tested the hypothesis that insulin therapy rather than sulfonylurea (SU) treatment is preferable to reverse or preserve beta-cell function among patients with slowly progressive insulin-dependent (type 1) diabetes (SPIDDM) or latent autoimmune diabetes in adults. METHODS: This multicenter, randomized, nonblinded clinical study screened 4089 non-insulin-dependent diabetic patients for glutamic acid decarboxylase autoantibodies (GADAb). Sixty GADAb-positive non insulin-requiring diabetic patients with a 5-yr duration or shorter of diabetes were assigned to either the SU group (n = 30) or the insulin group (n = 30). Serum C-peptide responses to annual oral glucose tolerance tests were followed up for a mean of 57 months. The primary endpoint was an insulin-dependent state defined by the sum of serum C-peptide values during the oral glucose tolerance test (SigmaC-peptide) less than 4 ng/ml (1.32 nmol/liter). RESULTS: The progression rate to an insulin-dependent state in the insulin group (three of 30, 10%) was lower than that in the SU group (13 of 30, 43%; P = 0.003, log-rank). Longitudinal analysis demonstrated that SigmaC-peptide values were better preserved in the insulin group than in the SU group. Multiple regression analysis demonstrated that insulin treatment, a preserved C-peptide response, and a low GADAb titer at entry were independent factors in preventing progression to an insulin-dependent state. Subgroup analysis suggested that insulin intervention was highly effective for SPIDDM patients with high GADAb titers [> or =10 U/ml (180 World Health Organization U/ml)] and preserved beta-cell function [SigmaC peptide > or = 10 ng/ml (3.31 nmol/liter)] at entry. No severe hypoglycemic episodes occurred during the study. CONCLUSIONS: Insulin intervention to preserve beta-cell function is effective and safe for patients with SPIDDM or latent autoimmune diabetes in adults. PMID- 18397987 TI - The aromatase inhibitor anastrozole is ineffective in the treatment of precocious puberty in girls with McCune-Albright syndrome. AB - CONTEXT: Precocious puberty (PP) in girls with McCune-Albright syndrome (MAS) is characterized by episodic development of large unilateral ovarian cysts followed by sudden onset of vaginal bleeding. Some patients experience frequent bleeding as well as accelerated linear growth and advanced skeletal maturation. The use of anastrozole for the treatment of PP in this condition has not been well studied. OBJECTIVE: The objective of the study was to determine the safety and efficacy of the aromatase inhibitor anastrozole for the treatment of PP in girls with MAS. DESIGN AND SETTINGS: This was a prospective international multicenter study in which subjects received anastrozole 1 mg daily for 1 yr. PATIENTS: Twenty-eight girls 10 years of age or younger with MAS and progressive PP were enrolled. MAIN OUTCOME MEASURES: Vaginal bleeding, rate of skeletal maturation (change in bone age over change in chronological age), growth velocity, and uterine/ovarian volumes were measured. These indices were compared with a 6-month pretreatment interval. RESULTS: No difference in vaginal bleeding (mean number of days per year) was noted. Mean change in DeltaBA/DeltaCA, which was 1.25 +/- 0.77 at baseline, was -0.25 +/- 1.02 at study end (P = 0.22). Average growth velocity z score was 1.40 +/- 3.15 at study entry and 0.26 +/- 2.71 at 12 months (P = 0.10). Mean ovarian/uterine volumes were unaffected by anastrozole, and no significant adverse events occurred. CONCLUSIONS: Although it appears safe, anastrozole for 1 yr was ineffective in halting vaginal bleeding, attenuating rates of skeletal maturation, and linear growth in girls with MAS. Pharmacological strategies other than anastrozole should be pursued for the treatment of PP in this population. PMID- 18397988 TI - Insulin sensitivity is correlated with subcutaneous but not visceral body fat in overweight and obese prepubertal children. AB - AIM: Our aim was to explore the relationship between insulin sensitivity, body fat distribution, ectopic (liver and skeletal muscle) fat deposition, adipokines (leptin and adiponectin), and inflammation markers (highly sensitive C-reactive protein, IL-6, IL-10, and TNF-alpha) in prepubertal children. SUBJECTS AND METHODS: Thirty overweight and obese children (16 males and 14 females with body mass index z-score range of 1.1-3.2) were recruited. Body fat distribution and fat accumulation in liver and skeletal muscle were measured using magnetic resonance imaging. Insulin sensitivity was assessed by iv glucose tolerance test. RESULTS: Insulin sensitivity was associated with sc abdominal adipose tissue (SAT) (r = -0.52; P < 0.01) and liver fat content (r = -0.44; P < 0.02) but not with visceral abdominal adipose tissue (VAT) (r = -0.193; P value not significant) and fat accumulation in skeletal muscle (r = -0.210; P value not significant). Adipokines, but not inflammation markers, were significantly correlated to insulin sensitivity. VAT correlated with C-reactive protein (r = 0.55; P < 0.01) as well as adiponectin (r = -0.53; P <0.01). Multiple regression analysis showed that only SAT and liver fat content were independently correlated to insulin sensitivity (P < 0.01; 20 and 16% of explained variance, respectively). CONCLUSIONS: In overweight and moderately obese prepubertal children, insulin sensitivity was negatively correlated with SAT and liver fat content. Furthermore, contrary to adults, VAT and inflammation markers were not correlated with insulin sensitivity in children. PMID- 18397990 TI - Physical and property victimization behind bars: a multilevel examination. AB - The majority of the extant literature on inmate victimization considers only one level of analysis, thus ignoring the interaction effects between inmate- and prison-level variables. To extend this literature, multilevel modeling techniques were used to analyze self-report data from more than 1,000 inmates and 30 prisons in Kentucky, Tennessee, and Ohio. Results revealed that demographic variables were strong predictors of physical victimization (i.e., race and assaultive behavior). Also, security level had a contextual direct effect on physical victimization. Property victimization was best explained with an integrated model including inmate (i.e., race, assaultive behavior, prior education, prior employment, and time served), contextual (i.e., security level and proportion non White), and micro-macro interaction variables (i.e., Race x Security Level). Policy implications and suggestions for future research are discussed. PMID- 18397989 TI - Leptin and ghrelin in relation to Helicobacter pylori status in adult males. AB - CONTEXT: Leptin and ghrelin, hormones involved in human energy homeostasis, are both produced in the stomach. OBJECTIVE: We sought to determine whether the presence of Helicobacter pylori affects gastric and systemic levels of leptin and ghrelin. DESIGN, SETTING, AND PATIENTS: We consecutively enrolled 256 patients referred for upper endoscopy at a Veterans Affairs outpatient endoscopy center. OUTCOMES: We obtained fasting serum, fundic and antral biopsies, and gastric juice. Based on histological, biochemical, and serological assays, patients were categorized as H. pylori+ or H. pylori-. Leptin and total ghrelin levels in serum, gastric biopsies, and gastric juice were determined by specific ELISAs. RESULTS: Of the 256 subjects, 120 were H. pylori+ and 96 were H. pylori-; 40 patients of indeterminate status were excluded. Serum and fundic leptin levels correlated with body mass index in the H. pylori+ (r = 0.35; P < 0.0001 and r = 0.35; P < 0.0001, respectively) and H. pylori- (r = 0.65; P < 0.0001 and r = 0.41; P < 0.0001, respectively) groups, but H. pylori+ subjects had significantly lower serum leptin levels [median 2.2 ng/ml (interquartile range 0.9-4.6) vs. 4.0 ng/ml (1.7-7.2); P = 0.0003]. Serum ghrelin levels were similar in the H. pylori+ and H. pylori- groups [median 1651 pg/ml (interquartile range 845-2247) vs. 1629 pg/ml (992-2886); P = 0.23]. H. pylori status did not significantly affect gastric biopsy leptin and ghrelin levels. Ghrelin levels in gastric juice varied over 4 log(10) (<80-776,000 pg/ml) and correlated with gastric juice pH in the H. pylori+ group (r = 0.68; P < 0.0001). CONCLUSIONS: These findings provide evidence that H. pylori status affects leptin and ghrelin homeostasis, presumably via intragastric interactions. PMID- 18397991 TI - Rape myth acceptance in men who completed the prostitution offender program of British Columbia. AB - In an effort to characterize the attitudes and characteristics of men who solicit sex, this study investigated rape myth acceptance as assessed by a modification of Burt's Rape Myth Acceptance Scale. The participants were all men who took part in the Prostitution Offender Program of British Columbia after being arrested for attempting to solicit sex from an undercover police officer. Relationships between endorsement of rape myths, other attitudes, sexual behavior, and demographic variables were examined. Results reveal that age, education, use of pornography, ideal frequency of intercourse, and believing that purchasing sex is a problem are all negatively correlated with rape myth acceptance. Positive correlations were found between rape myth acceptance and sexual conservatism, sexual violence/coercion, and social desirability. Results are discussed in terms of the association between rape myth acceptance and the violence frequently perpetrated against those working in the sex trade. PMID- 18397992 TI - A meta-analysis of QTL for diabetes-related traits in rodents. AB - Crossbreeding studies in rodents have identified numerous quantitative trait loci (QTL) that are linked to diabetes-related component traits. To identify genetic consensus regions implicated in insulin action and glucose homeostasis, we have performed a meta-analysis of genomewide linkage scans for diabetes-related traits. From a total of 43 published genomewide scans we assembled a nonredundant collection of 153 QTL for glucose levels, insulin levels, and glucose tolerance. Collectively, these studies include data from 48 different parental strains and >11,000 individual animals. The results of the studies were analyzed by the truncated product method (TPM). The analysis revealed significant evidence for linkage of glucose levels, insulin levels, and glucose tolerance to 27 different segments of the mouse genome. The most prominent consensus regions [localized to chromosomes 2, 4, 7, 9, 11, 13, and 19; logarithm of odds (LOD) scores 10.5-17.4] cover approximately 11% of the mouse genome and collectively contain the peak markers for 47 QTL. Approximately half of these genomic segments also show significant linkage to body weight and adiposity, indicating the presence of multiple obesity-dependent and -independent consensus regions for diabetes related traits. At least 84 human genetic markers from genomewide scans and >80 candidate genes from human and rodent studies map into the mouse consensus regions for diabetes-related traits, indicating a substantial overlap between the species. Our results provide guidance for the identification of novel candidate genes and demonstrate the presence of numerous distinct consensus QTL regions with highly significant LOD scores that control glucose homeostasis. An interactive physical map of the QTL is available online at http://www.diabesitygenes.org. PMID- 18397993 TI - beta-Catenin promotes self-renewal of skeletal-muscle satellite cells. AB - Satellite cells are the resident stem cells of adult skeletal muscle. As with all stem cells, how the choice between self-renewal or differentiation is controlled is central to understanding their function. Here, we have explored the role of beta-catenin in determining the fate of myogenic satellite cells. Satellite cells express beta-catenin, and expression is maintained as they activate and undergo proliferation. Constitutive retroviral-driven expression of wild-type or stabilised beta-catenin results in more satellite cells expressing Pax7 without any MyoD -- therefore, adopting the self-renewal pathway, with fewer cells undergoing myogenic differentiation. Similarly, preventing the degradation of endogenous beta-catenin by inhibiting GSK3beta activity also results in more Pax7 positive-MyoD-negative (Pax7(+)MyoD(-)) satellite-cell progeny. Consistent with these observations, downregulation of beta-catenin using small interfering RNA (siRNA) reduced the proportion of satellite cells that express Pax7 and augmented myogenic differentiation after mitogen withdrawal. Since a dominant-negative version of beta-catenin had the same effect as silencing beta-catenin using specific siRNA, beta-catenin promotes self-renewal via transcriptional control of target genes. Thus, beta-catenin signalling in proliferating satellite cells directs these cells towards the self-renewal pathway and, so, contributes to the maintenance of this stem-cell pool in adult skeletal muscle. PMID- 18397994 TI - Slk1 is a meiosis-specific Sid2-related kinase that coordinates meiotic nuclear division with growth of the forespore membrane. AB - Septation and spore formation in fission yeast are compartmentalization processes that occur during the mitotic and meiotic cycles, and that are regulated by the septation initiation network (SIN). In mitosis, activation of Sid2 protein kinase transduces the signal from the spindle pole body (SPB) to the middle of the cell in order to promote the constriction of the actomyosin ring. Concomitant with ring contraction, membrane vesicles are added at the cleavage site to enable the necessary expansion of the cell membrane. In meiosis, the forespore membrane is synthesized from the outer layers of the SPB by vesicle fusion. This membrane grows and eventually engulfs each of the four haploid nuclei. The molecular mechanism that connects the SIN pathway with synthesis of the forespore membrane is poorly understood. Here, we describe a meiosis-specific Sid2-like kinase (Slk1), which is important for the coordination of the growth of the forespore membrane with the meiotic nuclear divisions. Slk1 and Sid2 are required for forespore membrane biosynthesis and seem to be the final output of the SIN pathway in meiosis. PMID- 18397995 TI - Fibroblast migration is mediated by CD44-dependent TGF beta activation. AB - CD44 contributes to inflammation and fibrosis in response to injury. As fibroblast recruitment is critical to wound healing, we compared cytoskeletal architecture and migration of wild-type (CD44WT) and CD44-deficient (CD44KO) fibroblasts. CD44KO fibroblasts exhibited fewer stress fibers and focal adhesion complexes, and their migration was characterized by increased velocity but loss of directionality, compared with CD44WT fibroblasts. Mechanistically, we demonstrate that CD44WT cells generated more active TGFbeta than CD44KO cells and that CD44 promotes the activation of TGFbeta via an MMP-dependent mechanism. Reconstitution of CD44 expression completely rescued the phenotype of CD44KO cells whereas exposure of CD44KO cells to exogenous active TGFbeta rescued the defect in stress fibers and migrational velocity, but was not sufficient to restore directionality of migration. These results resolve the TGFbeta-mediated and TGFbeta-independent effects of CD44 on fibroblast migration and suggest that CD44 may be critical for the recruitment of fibroblasts to sites of injury and the function of fibroblasts in tissue remodeling and fibrosis. PMID- 18397997 TI - alphaE-catenin is not a significant regulator of beta-catenin signaling in the developing mammalian brain. AB - beta-Catenin is a crucial mediator of the canonical Wnt-signaling pathway. alpha catenin is a major beta-catenin-binding protein, and overexpressed alpha-catenin can negatively regulate beta-catenin activity. Thus, alpha-catenin may be an important modulator of the Wnt pathway. We show here that endogenous alpha catenin has little impact on the transcriptional activity of beta-catenin in developing mammalian organisms. We analyzed beta-catenin signaling in mice with conditional deletion of alphaE-catenin (Ctnna1) in the developing central nervous system. This mutation results in brain hyperplasia and we investigated whether activation of beta-catenin signaling may be at least partially responsible for this phenotype. To reveal potential quantitative or spatial changes in beta catenin signaling, we used mice carrying a beta-catenin-signaling reporter transgene. In addition, we analyzed the expression of known endogenous targets of the beta-catenin pathway and the amount and localization of beta-catenin in mutant progenitor cells. We found that although loss of alphaE-catenin resulted in disruption of intercellular adhesion and hyperplasia in the developing brain, beta-catenin signaling was not altered. We conclude that endogenous alphaE catenin has no significant impact on beta-catenin transcriptional activities in the developing mammalian brain. PMID- 18397996 TI - Bone marrow side population cells are enriched for progenitors capable of myogenic differentiation. AB - Although the contribution of bone marrow-derived cells to regenerating skeletal muscle has been repeatedly documented, there remains considerable debate as to whether this incorporation is exclusively a result of inflammatory cell fusion to regenerating myofibers or whether certain populations of bone marrow-derived cells have the capacity to differentiate into muscle. The present study uses a dual-marker approach in which GFP(+) cells were intravenously transplanted into lethally irradiated beta-galactosidase(+) recipients to allow for simple determination of donor and host contribution to the muscle. FACS analysis of cardiotoxin-damaged muscle revealed that CD45(+) bone-marrow side-population (SP) cells, a group enriched in hematopoietic stem cells, can give rise to CD45(-)/Sca 1(+)/desmin(+) cells capable of myogenic differentiation. Moreover, after immunohistochemical examination of the muscles of both SP- and whole bone marrow transplanted animals, we noted the presence of myofibers composed only of bone marrow-derived cells. Our findings suggest that a subpopulation of bone marrow SP cells contains precursor cells whose progeny have the potential to differentiate towards a muscle lineage and are capable of de novo myogenesis following transplantation and initiation of muscle repair via chemical damage. PMID- 18397998 TI - Inhibition of beta-catenin signaling causes defects in postnatal cartilage development. AB - The Wnt/beta-catenin signaling pathway is essential for normal skeletal development because conditional gain or loss of function of beta-catenin in cartilage results in embryonic or early postnatal death. To address the role of beta-catenin in postnatal skeletal growth and development, Col2a1-ICAT transgenic mice were generated. Mice were viable and had normal size at birth, but became progressively runted. Transgene expression was limited to the chondrocytes in the growth plate and articular cartilages and was associated with decreased beta catenin signaling. Col2a1-ICAT transgenic mice showed reduced chondrocyte proliferation and differentiation, and an increase in chondrocyte apoptosis, leading to decreased widths of the proliferating and hypertrophic zones, delayed formation of the secondary ossification center, and reduced skeletal growth. Isolated primary Col2a1-ICAT transgenic chondrocytes showed reduced expression of chondrocyte genes associated with maturation, and demonstrated that VEGF gene expression requires cooperative interactions between BMP2 and beta-catenin signaling. Altogether the findings confirm a crucial role for Wnt/beta-catenin in postnatal growth. PMID- 18397999 TI - Caveolin-1 alters Ca(2+) signal duration through specific interaction with the G alpha q family of G proteins. AB - Caveolae are membrane domains having caveolin-1 (Cav1) as their main structural component. Here, we determined whether Cav1 affects Ca(2+) signaling through the Galpha(q)-phospholipase-Cbeta (PLCbeta) pathway using Fischer rat thyroid cells that lack Cav1 (FRTcav(-)) and a sister line that forms caveolae-like domains due to stable transfection with Cav1 (FRTcav(+)). In the resting state, we found that eCFP-Gbetagamma and Galpha(q)-eYFP are similarly associated in both cell lines by Forster resonance energy transfer (FRET). Upon stimulation, the amount of FRET between Galpha(q)-eYFP and eCFP-Gbetagamma remains high in FRTcav(-) cells, but decreases almost completely in FRTcav(+) cells, suggesting that Cav1 is increasing the separation between Galpha(q)-Gbetagamma subunits. In FRTcav(-) cells overexpressing PLCbeta, a rapid recovery of Ca(2+) is observed after stimulation. However, FRTcav(+) cells show a sustained level of elevated Ca(2+). FRET and colocalization show specific interactions between Galpha(q) and Cav1 that increase upon stimulation. Fluorescence correlation spectroscopy studies show that the mobility of Galpha(q)-eGFP is unaffected by activation in either cell type. The mobility of eGFP-Gbetagamma remains slow in FRTcav(-) cells but increases in FRTcav(+) cells. Together, our data suggest that, upon stimulation, Galpha(q)(GTP) switches from having strong interactions with Gbetagamma to Cav1, thereby releasing Gbetagamma. This prolongs the recombination time for the heterotrimer, thus causing a sustained Ca(2+) signal. PMID- 18398000 TI - Asymmetric localization of the adaptor protein Miranda in neuroblasts is achieved by diffusion and sequential interaction of Myosin II and VI. AB - The adaptor protein Miranda plays a pivotal role in the asymmetric cell division of neuroblasts by asymmetrically segregating key differentiation factors. Miranda localization requires Myosin VI and Myosin II. The apical-then-basal localization pattern of Miranda detected in fixed tissue, and the localization defects in embryos lacking Myosin VI, suggest that Miranda is transported to the basal pole as a Myosin VI cargo. However, the mode and temporal sequence of Miranda localization have not been characterized in live embryos. Furthermore, it is unknown whether Miranda and PON, a second adaptor protein required for asymmetric protein localization, are both regulated by Myosin II. By combining immunofluorescence studies with time-lapse confocal microscopy, we show that Miranda protein forms an apical crescent at interphase, but is ubiquitously localized at prophase in a Myosin-II-dependent manner. FRAP analysis revealed that Miranda protein reaches the basal cortex by passive diffusion throughout the cell, rather than by long-range Myosin VI-directed transport. Myosin VI acts downstream of Myosin II in the same pathway to deliver diffusing Miranda to the basal cortex. PON localization occurs mainly along the cortex and requires Myosin II but not Myosin VI, suggesting that distinct mechanisms are employed to localize different adaptor proteins during asymmetric cell division. PMID- 18398001 TI - Control of thrombin signaling through PI3K is a mechanism underlying plasticity between hair follicle dermal sheath and papilla cells. AB - In hair follicles, dermal papilla (DP) and dermal sheath (DS) cells exhibit striking levels of plasticity, as each can regenerate both cell types. Here, we show that thrombin induces a phosphoinositide 3-kinase (PI3K)-Akt pathway dependent acquisition of DS-like properties by DP cells in vitro, involving increased proliferation rate, acquisition of ;myofibroblastic' contractile properties and a decreased capacity to sustain growth and survival of keratinocytes. The thrombin inhibitor protease nexin 1 [PN-1, also known as SERPINE2) regulates all those effects in vitro. Accordingly, the PI3K-Akt pathway is constitutively activated and expression of myofibroblastic marker smooth muscle actin is enhanced in vivo in hair follicle dermal cells from PN-1(-/-) mice. Furthermore, physiological PN-1 disappearance and upregulation of the thrombin receptor PAR-1 (also known as F2R) during follicular regression in wild type mice also correlate with such changes in DP cell characteristics. Our results indicate that control of thrombin signaling interferes with hair follicle dermal cells plasticity to regulate their function. PMID- 18398002 TI - Dynamics of an F-actin aggresome generated by the actin-stabilizing toxin jasplakinolide. AB - In this study, we report the formation of several cytoplasmic inclusion bodies composed of filamentous actin (F-actin) and generated by experimental treatments using depolymerizing or stabilizing actin toxins in neuronal and non-neuronal mammalian cell lines. The actin-stabilizing toxin jasplakinolide (Jpk) induced, in a microtubule-dependent manner, a single, large F-actin aggregate, which contained beta- and gamma-actin, ADF/cofilin, cortactin, and the actin nucleator Arp2/3. This aggregate was tightly associated with the Golgi complex and mitochondria, and was surrounded by vimentin intermediate filaments, microtubules and MAP4. Therefore, the Jpk-induced single, large F-actin aggregate fits the established criteria for being considered an aggresome. Lysosomes and/or autophagic vacuoles, proteasomes and microtubules were found to directly participate in the dissolution of this F-actin aggresome. Finally, the model reported here is simple, highly reproducible and reversible, and it provides an opportunity to test pharmacological agents that interfere with the formation, maintenance and/or disappearance of F-actin-enriched pathological inclusion bodies. PMID- 18398003 TI - Morphing peptide backbones into heterocycles. PMID- 18398004 TI - Artificial miRNAs mitigate shRNA-mediated toxicity in the brain: implications for the therapeutic development of RNAi. AB - Huntington's disease (HD) is a fatal, dominant neurodegenerative disease caused by a polyglutamine repeat expansion in exon 1 of the HD gene, which encodes the huntingtin protein. We and others have shown that RNAi is a candidate therapy for HD because expression of inhibitory RNAs targeting mutant human HD transgenes improved neuropathology and behavioral deficits in HD mouse models. Here, we developed shRNAs targeting conserved sequences in human HD and mouse HD homolog (HDh) mRNAs to initiate preclinical testing in a knockin mouse model of HD. We screened 35 shRNAs in vitro and subsequently narrowed our focus to three candidates for in vivo testing. Unexpectedly, two active shRNAs induced significant neurotoxicity in mouse striatum, although HDh mRNA expression was reduced to similar levels by all three. Additionally, a control shRNA containing mismatches also induced toxicity, although it did not reduce HDh mRNA expression. Interestingly, the toxic shRNAs generated higher antisense RNA levels, compared with the nontoxic shRNA. These results demonstrate that the robust levels of antisense RNAs emerging from shRNA expression systems can be problematic in the mouse brain. Importantly, when sequences that were toxic in the context of shRNAs were placed into artificial microRNA (miRNA) expression systems, molecular and neuropathological readouts of neurotoxicity were significantly attenuated without compromising mouse HDh silencing efficacy. Thus, miRNA-based approaches may provide more appropriate biological tools for expressing inhibitory RNAs in the brain, the implications of which are crucial to the development of RNAi for both basic biological and therapeutic applications. PMID- 18398005 TI - Fine structure mapping of a gene-rich region of wheat carrying Ph1, a suppressor of crossing over between homoeologous chromosomes. AB - The wheat gene-rich region (GRR) 5L0.5 contains many important genes, including Ph1, the principal regulator of chromosome pairing. Comparative marker analysis identified 32 genes for the GRR controlling important agronomic traits. Detailed characterization of this region was accomplished by first physically localizing 213 wheat group 5L-specific markers, using group 5 nulli-tetrasomics, three Ph1 gene deletion/insertion mutants, and nine terminal deletion lines with their breakpoints around the 5L0.5 region. The Ph1 gene was localized to a much smaller region within the GRR (Ph1 gene region). Of the 61 markers that mapped in the four subregions of the GRR, 9 mapped in the Ph1 gene region. High stringency sequence comparison (e < 1 x10(-25)) of 157 group 5L-specific wheat ESTs identified orthologs for 80% sequences in rice and 71% in Arabidopsis. Rice orthologs were present on all rice chromosomes, although most (34%) were on rice chromosome 9 (R9). No single collinear region was identified in Arabidopsis even for a smaller region, such as the Ph1 gene region. Seven of the nine Ph1 gene region markers mapped within a 450-kb region on R9 with the same gene order. Detailed domain/motif analysis of the 91 putative genes present in the 450-kb region identified 26 candidates for the Ph1 gene, including genes involved in chromatin reorganization, microtubule attachment, acetyltransferases, methyltransferases, DNA binding, and meiosis/anther specific proteins. Five of these genes shared common domains/motifs with the meiosis specific genes Zip1, Scp1, Cor1, RAD50, RAD51, and RAD57. Wheat and Arabidopsis homologs for these rice genes were identified. PMID- 18398006 TI - Bacterial addiction module toxin Doc inhibits translation elongation through its association with the 30S ribosomal subunit. AB - Bacterial toxin-antitoxin (TA) systems (or "addiction modules") typically facilitate cell survival during intervals of stress by inducing a state of reversible growth arrest. However, upon prolonged stress, TA toxin action leads to cell death. TA systems have also been implicated in several clinically important phenomena: biofilm formation, bacterial persistence during antibiotic treatment, and bacterial pathogenesis. TA systems harbored by pathogens also serve as attractive antibiotic targets. To date, the mechanism of action of the majority of known TA toxins has not yet been elucidated. We determined the mode of action of the Doc toxin of the Phd-Doc TA system. Doc expression resulted in rapid cell growth arrest and marked inhibition of translation without significant perturbation of transcription or replication. However, Doc did not cleave mRNA as do other addiction-module toxins whose activities result in translation inhibition. Instead, Doc induction mimicked the effects of treatment with the aminoglycoside antibiotic hygromycin B (HygB): Both Doc and HygB interacted with 30S ribosomal subunits, stabilized polysomes, and resulted in a significant increase in mRNA half-life. HygB also competed with ribosome-bound Doc, whereas HygB-resistant mutants suppressed Doc toxicity, suggesting that the Doc-binding site includes that of HygB (i.e., helix 44 region of 16S rRNA containing the A, P, and E sites). Overall, our results illuminate an intracellular target and mechanism of TA toxin action drawn from aminoglycoside antibiotics: Doc toxicity is the result of inhibition of translation elongation, possibly at the translocation step, through its interaction with the 30S ribosomal subunit. PMID- 18398007 TI - Molecular assembly for high-performance bivalent nucleic acid inhibitor. AB - It is theorized that multivalent interaction can result in better affinity and selectivity than monovalent interaction in the design of high-performance ligands. Accordingly, biomolecular engineers are increasingly taking advantage of multivalent interactions to fabricate novel molecular assemblies, resulting in new functions for ligands or enhanced performance of existing ligands. Substantial efforts have been expended in using small molecules or epitopes of antibodies for designing multifunctional or better-performing ligands. However, few attempts to use nucleic acid aptamers as functional domains have been reported. In this study, we explore the design of bivalent nucleic acid ligands by using thrombin and its aptamers as the model by which to evaluate its functions. By assembling two thrombin-binding aptamers with optimized design parameters, this assembly has resulted in the successful development of a nucleic acid-based high-performance bivalent protein inhibitor. Our experimentation proved (i) that the simultaneous binding of two aptamers after linkage achieved 16.6-fold better inhibition efficiency than binding of the monovalent ligand and (ii) that such an improvement originated from changes in the kinetics of the binding interactions, with a k(off) rate approximately 1/50 as fast. In addition, the newly generated aptamer assembly is an excellent anticoagulant reagent when tested with different samples. Because this optimized ligand design offers a simple and noninvasive means of accomplishing higher performance from known functional aptamers, it holds promise as a potent antithrombin agent in the treatment of various diseases related to abnormal thrombin activities. PMID- 18398008 TI - The catalytic scaffold of the haloalkanoic acid dehalogenase enzyme superfamily acts as a mold for the trigonal bipyramidal transition state. AB - The evolution of new catalytic activities and specificities within an enzyme superfamily requires the exploration of sequence space for adaptation to a new substrate with retention of those elements required to stabilize key intermediates/transition states. Here, we propose that core residues in the large enzyme family, the haloalkanoic acid dehalogenase enzyme superfamily (HADSF) form a "mold" in which the trigonal bipyramidal transition states formed during phosphoryl transfer are stabilized by electrostatic forces. The vanadate complex of the hexose phosphate phosphatase BT4131 from Bacteroides thetaiotaomicron VPI 5482 (HPP) determined at 1.00 A resolution via X-ray crystallography assumes a trigonal bipyramidal coordination geometry with the nucleophilic Asp-8 and one oxygen ligand at the apical position. Remarkably, the tungstate in the complex determined to 1.03 A resolution assumes the same coordination geometry. The contribution of the general acid/base residue Asp-10 in the stabilization of the trigonal bipyramidal species via hydrogen-bond formation with the apical oxygen atom is evidenced by the 1.52 A structure of the D10A mutant bound to vanadate. This structure shows a collapse of the trigonal bipyramidal geometry with displacement of the water molecule formerly occupying the apical position. Furthermore, the 1.07 A resolution structure of the D10A mutant complexed with tungstate shows the tungstate to be in a typical "phosphate-like" tetrahedral configuration. The analysis of 12 liganded HADSF structures deposited in the protein data bank (PDB) identified stringently conserved elements that stabilize the trigonal bipyramidal transition states by engaging in favorable electrostatic interactions with the axial and equatorial atoms of the transferring phosphoryl group. PMID- 18398009 TI - Saturation mutagenesis of putative catalytic residues of benzoylformate decarboxylase provides a challenge to the accepted mechanism. AB - Benzoylformate decarboxylase from Pseudomonas putida (PpBFDC) is a thiamin diphosphate-dependent enzyme that carries out the nonoxidative decarboxylation of aromatic 2-keto acids. The x-ray structure of PpBFDC suggested that Ser-26, His 70, and His-281 would play important roles in its catalytic mechanism, and the S26A, H70A, and H281A variants all exhibited greatly impaired catalytic activity. Based on stopped-flow studies with the alanine mutants, it was proposed that the histidine residues acted as acid-base catalysts, whereas Ser-26 was involved in substrate binding and played a significant, albeit less well defined, role in catalysis. While developing a saturation mutagenesis protocol to examine residues involved in PpBFDC substrate specificity, we tested the procedure on His-281. To our surprise, we found that His-281, which is thought to be necessary for protonation of the carbanion/enamine intermediate, could be replaced by phenyl alanine with only a 5-fold decrease in k(cat). Even more surprising were our subsequent observations (i) that His-70 could be replaced by threonine or leucine with approximately a 30-fold decrease in k(cat)/K(m) compared with a 4,000-fold decrease for the H70A variant and (ii) that Ser-26, which forms hydrogen bonds with the substrate carboxylate, could be replaced by threonine, leucine, or methionine without significant loss of activity. These results call into question the assigned roles for Ser-26, His-70, and His-281. Further, they demonstrate the danger in assigning catalytic function based solely on results with alanine mutants and show that saturation mutagenesis is a valuable tool in assessing the role and relative importance of putative catalytic residues. PMID- 18398010 TI - A review of Parkinson's disease. AB - INTRODUCTION: Parkinson's disease (PD) is one of the most common neurodegenerative disorders. Sources of data Literature search using Medline with keywords Parkinson's disease supplemented with previously published papers known to the author. AREAS OF AGREEMENT: There have been significant recent advances in the understanding of the pathogenesis of the disease. There has also been a greater realization that the disorder may be associated with significant non motor disturbances in addition to the more commonly recognized motor complications. AREAS OF CONTROVERSY: Although there is growing circumstantial evidence, it remains to be proven whether any of the current treatments for PD have a neuroprotective effect. AREAS TIMELY FOR DEVELOPING RESEARCH: Although there is no cure, there are several management options for the early treatment of PD. As the disease progresses, further treatment options are available; however, the management of late-stage motor complications and non-motor symptoms remains particularly challenging and will benefit from further clinical research. PMID- 18398012 TI - Shift in metabolic fuel in acylation-stimulating protein-deficient mice following a high-fat diet. AB - ASP-deficient mice (C3 KO) have delayed postprandial TG clearance, are hyperphagic, and display increased energy expenditure. Markers of carbohydrate and fatty acid metabolism in the skeletal muscle and heart were examined to evaluate the mechanism. On a high-fat diet, compared with wild-type mice, C3 KO mice have increased energy expenditure, decreased RQ, lower ex vivo glucose oxidation (-39%, P = 0.018), and higher ex vivo fatty acid oxidation (+68%, P = 0.019). They have lower muscle glycogen content (-25%, P < 0.05) and lower activities for the glycolytic enzymes glycogen phosphorylase (-31%, P = 0.005), hexokinase (-43%, P = 0.007), phosphofructokinase (-51%, P < 0.0001), and GAPDH ( 15%, P = 0.04). Analysis of mitochondrial enzyme activities revealed that hydroxyacyl-coenzyme A dehydrogenase was higher (+25%, P = 0.004) in C3 KO mice. Furthermore, Western blot analysis of muscle revealed significantly higher fatty acid transporter CD36 (+40%, P = 0.006) and cytochrome c (a marker of mitochondrial content; +69%, P = 0.034) levels in C3 KO mice, whereas the activity of AMP kinase was lower (-48%, P = 0.003). Overall, these results demonstrate a shift in the metabolic potential of skeletal muscle toward increased fatty acid utilization. Whether this is 1) a consequence of decreased adipose tissue storage with repartitioning toward muscle or 2) a direct result of the absence of ASP interaction with the receptor C5L2 in muscle remains to be determined. However, these in vivo data suggest that ASP inhibition could be a potentially viable approach in correcting muscle metabolic dysfunction in obesity. PMID- 18398014 TI - Utilization of nanobiotechnology in haemodialysis: mock-dialysis experiments on homocysteine. AB - BACKGROUND: The utilization of modern achievements from nanobiotechnology has resulted in novel modalities for renal replacement therapy. For conventional intermittent haemodialysis (HD), sophisticated membranes are currently being manufactured that guarantee selective removal of target toxins. These membranes have a narrow pore-size distribution that is focused around a mean value at the nanometre level. For continuous HD, novel artificial renal devices are currently being designed and evaluated in in vitro experiments that will be both implantable and have continuous function. METHODS: We present mock-dialysis experiments using magnetically assisted HD (MAHD) that we very recently introduced for the selective removal of target toxins. MAHD is based on the preparation of conjugates (Cs) made up of biocompatible ferromagnetic nanoparticles (FNs) and a specifically designed targeted binding substance that must have a high affinity for a specific target toxin substance. The FN-targeted binding substance Cs should be administered to the patient prior to MAHD to allow for binding with the target toxin substance in the bloodstream. The complex FN targeted binding substance-target toxin substance will then be removed by a 'magnetic dialyzer' that is installed in the dialysis machine in series to the conventional dialyzer. In the present work, we compared the in vitro efficiency of MAHD to conventional HD for the removal of homocysteine (Hcy) during mock dialysis experiments. RESULTS: These mock-dialysis experiments performed on Hcy revealed that both the removal rate and the overall removal efficiency of MAHD were significantly greater than conventional HD. CONCLUSIONS: MAHD appears to be a promising method that can be employed for the selective and more efficient extraction of toxins that are not adequately removed by conventional HD. PMID- 18398013 TI - Vestibulosympathetic reflex during the early follicular and midluteal phases of the menstrual cycle. AB - Evidence suggests that both the arterial baroreflex and vestibulosympathetic reflex contribute to blood pressure regulation, and both autonomic reflexes integrate centrally in the medulla cardiovascular center. A previous report indicated increased sympathetic baroreflex sensitivity during the midluteal (ML) phase of the menstrual cycle compared with the early follicular (EF) phase. On the basis of this finding, we hypothesize an augmented vestibulosympathetic reflex during the ML phase of the menstrual cycle. Muscle sympathetic nerve activity (MSNA), mean arterial pressure (MAP), and heart rate responses to head down rotation (HDR) were measured in 10 healthy females during the EF and ML phases of the menstrual cycle. Plasma estradiol (Delta72 +/- 13 pg/ml, P < 0.01) and progesterone (Delta8 +/- 2 ng/ml, P < 0.01) were significantly greater during the ML phase compared with the EF phase. The menstrual cycle did not alter resting MSNA, MAP, and heart rate (EF: 13 +/- 3 bursts/min, 80 +/- 2 mmHg, 65 +/- 2 beats/min vs. ML: 14 +/- 3 bursts/min, 81 +/- 3 mmHg, 64 +/- 3 beats/min). During the EF phase, HDR increased MSNA (Delta3 +/- 1 bursts/min, P < 0.02) but did not change MAP or heart rate (Delta0 +/- 1 mmHg and Delta1 +/- 1 beats/min). During the ML phase, HDR increased both MSNA and MAP (Delta4 +/- 1 bursts/min and Delta3 +/- 1 mmHg, P < 0.04) with no change in heart rate (Delta0 +/- 1 beats/min). MSNA and heart rate responses to HDR were not different between the EF and ML phases, but MAP responses to HDR were augmented during the ML phase (P < 0.03). Our results demonstrate that the menstrual cycle does not influence the vestibulosympathetic reflex but appears to alter MAP responses to HDR during the ML phase. PMID- 18398011 TI - Regulation of the fructose transporter GLUT5 in health and disease. AB - Fructose is now such an important component of human diets that increasing attention is being focused on the fructose transporter GLUT5. In this review, we describe the regulation of GLUT5 not only in the intestine and testis, where it was first discovered, but also in the kidney, skeletal muscle, fat tissue, and brain where increasing numbers of cell types have been found to have GLUT5. GLUT5 expression levels and fructose uptake rates are also significantly affected by diabetes, hypertension, obesity, and inflammation and seem to be induced during carcinogenesis, particularly in the mammary glands. We end by highlighting research areas that should yield information needed to better understand the role of GLUT5 during normal development, metabolic disturbances, and cancer. PMID- 18398015 TI - Cancer in patients on dialysis and after renal transplantation. PMID- 18398016 TI - Evaluation of parathyroid gland angiogenesis in chronic kidney disease associated with secondary hyperparathyroidism. AB - BACKGROUND: Secondary hyperparathyroidism (SHPT) is a common complication of chronic kidney disease. Increased parathyroid hormone (PTH) synthesis and secretion is associated with parathyroid cell hyperplasia. The exact mechanisms involved in parathyroid gland (PTG) hyperplasia are still poorly understood. There is no available data on angiogenesis in PTG of patients with chronic kidney disease and SHPT. The aim of this study is to evaluate angiogenesis and expression of the angiogenic factors, basic fibroblastic growth factor (b-FGF) and vascular endothelial growth factor A (VEGF), in secondary PTG hyperplasia. METHODS: This study was performed on formalin-fixed paraffin-embedded archival tissues of 21 SHPT glands from haemodialysis (n = 19) and kidney transplanted (n = 2) patients submitted to surgical parathyroidectomy. For control, eight normal human parathyroid glands (NPG) encountered in surgical specimens of total thyroidectomy were used. We evaluated the immunohistochemical expression of the proliferation cell marker Ki67. Angiogenesis was evaluated by immunohistochemistry staining with anti-endoglin (CD105) antibody in 21 SPH and 5 NPG by stereological analysis. Levels of b-FGF and VEGF were determined by semi quantitative analysis in 21 SPH and 8 NPG. RESULTS: The SHPT patients present a mean iPTH of 1314 +/- 750 pg/ml, a corrected serum calcium of 10.3 +/- 1.2 mg/dl and a serum phosphorus of 6.1 +/- 1.4 mg/dl. SHPT glands displayed a significantly higher immunoreactivity against Ki67, compared to NPG. With CD105, a significantly higher number and volume of microvessels were observed in SHPT compared to NPG. Both VEGF and b-FGF expression were increased in SHPT compared to NPG. Using the predefined subdivision into negative and positive only the b FGF expression was significantly increased in the SHPT glands compared to NPG. CONCLUSION: These results suggest that PTGs in this group of patients with SHPT have a significantly higher number of vessels expressing CD105, which has been reported to preferentially label activated endothelial cells associated with angiogenesis. SHPT glands have a significantly increased expression of b-FGF compared to NPG. VEGF-A expression is also increased in the examined SHPT glands but could be less relevant for angiogenesis. PMID- 18398017 TI - Protein adsorption during LDL-apheresis: proteomic analysis. AB - BACKGROUND: The aim of our study was to investigate the clearance of functional proteins by different low-density lipoprotein-apheresis (LDL-A) methods with the help of proteomic analyses. METHODS: Proteins were eluated from the different LDL A columns and investigated with 2D electrophoresis combined with mass spectrometry methods. In parallel, we quantified the plasma protein loss from patients treated with double-filtration plasmapheresis (DFPP; n = 9), direct adsorption of lipoproteins (DALI; n = 5) or heparin-induced extracorporeal LDL precipitation (HELP; n = 7) with routine laboratory methods and western blots. RESULTS: Proteomic analyses of the column-bound proteins revealed a column-type dependent loss with the highest number of protein spots in DALI-treated patients (1001 +/- 36), followed by HELP (881 +/- 25) and DFPP (535 +/- 20). More than 70 functional proteins were identified. These proteins are involved in the coagulation pathway (e.g. kininogen1) and have adhesive (e.g. fibronectin), rheological (e.g. fibrinogen) and immunological/inflammatory properties (e.g. complement components). Quantification with western blot analyses demonstrated a significant depletion (P < 0.01) of these proteins comparing serum samples before and after the column with a systemic lowering in patients' serum. CONCLUSIONS: These data reveal strong interaction between column and serum proteins during LDL A. The clearance of proteins with adhesive, rheological, and inflammatory characteristics may have beneficial effects on microcirculation and reduce chronic inflammation but may also concomitantly induce side effects such as an increased bleeding risk. PMID- 18398018 TI - Kidney function decline and physical function in women. AB - BACKGROUND: Cross-sectional analyses of kidney function and physical function have identified profound quality of life impairments in people with advanced kidney dysfunction. No data are currently available, however, on how kidney function decline may be associated with physical function. METHODS: We undertook a study of kidney function decline and physical function in 2544 women participating in the Nurses' Health Study. Glomerular filtration rates (GFR) were estimated using the four-variable MDRD equation from plasma creatinine measured in blood collected in 1989 and 2000. Physical function was assessed by the Physical Function Sub-Scale (PFS) score of the Short Form 36 (SF-36) in a questionnaire administered in the year 2000. PFS scores have been shown to correlate well with direct measures of physical function. RESULTS: In the year 2000, the median age was 67 years, median body mass index (BMI) was 25.6 kg/m(2), 48.5% had hypertension and 5.8% had diabetes. There were 427 women (16.8%) who experienced an >/=25% decline in eGFR between 1989 and 2000. Median PFS in 2000 for those with an eGFR decline of >/=25% was 80 compared to a PFS score of 85 for those without (P < 0.001). In fully adjusted models, the presence of an eGFR decline of >/=25% was independently associated with a 3.5-point lower PFS score (95% CI -5.4 to -1.5). Also, an eGFR decline of >/=25% was independently associated with an increased odds ratio of being in the lowest quartile of PFS score (OR 1.37; 95% CI 1.04-1.81). CONCLUSIONS: We conclude that an eGFR decline of >/=25% over 11 years is independently associated with lower physical function in women. PMID- 18398019 TI - PTH and 1.25 vitamin D response to a low-calcium diet is associated with bone mineral density in renal stone formers. AB - BACKGROUND: Renal calcium stones and hypercalciuria are associated with a reduced bone mineral density (BMD). Therefore, the effect of changes in calcium homeostasis is of interest for both stones and bones. We hypothesized that the response of calciuria, parathyroid hormone (PTH) and 1.25 vitamin D to changes in dietary calcium might be related to BMD. METHODS: A single-centre prospective interventional study of 94 hyper- and non-hypercalciuric calcium stone formers consecutively retrieved from our stone clinic. The patients were investigated on a free-choice diet, a low-calcium diet, while fasting and after an oral calcium load. Patient groups were defined according to lumbar BMD (z-score) obtained by dual X-ray absorptiometry (group 1: z-score <-0.5, n = 30; group 2: z-score -0.5 0.5, n = 36; group 3: z-score >0.5, n = 28). The effect of the dietary interventions on calciuria, 1.25 vitamin D and PTH in relation to BMD was measured. RESULTS: An inverse relationship between BMD and calciuria was observed on all four calcium intakes (P = 0.009). On a free-choice diet, 1.25 vitamin D and PTH levels were identical in the three patient groups. However, the relative responses of 1.25 vitamin D and PTH to the low-calcium diet were opposite in the three groups with the highest increase of 1.25 vitamin D in group 1 and the lowest in group 3, whereas PTH increase was most pronounced in group 3 and least in group 1. CONCLUSION: Calcium stone formers with a low lumbar BMD exhibit a blunted response of PTH release and an apparently overshooting production of 1.25 vitamin D following a low-calcium diet. PMID- 18398020 TI - The case for a gender-neutral (universal) human papillomavirus vaccination policy in the United States: Point. PMID- 18398021 TI - Human papillomavirus vaccination should be offered to young males: Counterpoint. PMID- 18398022 TI - Human papillomavirus in cervical cancer screening: important role as biomarker. AB - Cervical cytology screening has reduced cervical cancer morbidity and mortality but shows important shortcomings in terms of sensitivity and specificity. Infection with distinct types of human papillomavirus (HPV) is the primary etiologic factor in cervical carcinogenesis. This causal relationship has been exploited for the development of molecular technologies for viral detection to overcome limitations linked to cytologic cervical screening. HPV testing has been suggested for primary screening, triage of equivocal Pap smears or low-grade lesions and follow-up after treatment for cervical intraepithelial neoplasia. Determination of HPV genotype, viral load, integration status and RNA expression could further improve the effectiveness of HPV-based screening and triage strategies. The prospect of prophylactic HPV vaccination stresses the importance of modification of the current cytology-based screening approach. PMID- 18398023 TI - Recruitment in the prostate, lung, colorectal, and ovarian (PLCO) cancer screening trial: the first phase of recruitment at Henry Ford Health System. AB - OBJECTIVE: Recruitment of healthy subjects to long-term randomized controlled trials (RCTs) of cancer prevention or early detection has proven to be a difficult task. To quantify recruitment yield as well as characteristics of successfully recruited participants, we examined recruitment outcomes at 1 of the 10 centers participating in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial, a National Cancer Institute-funded RCT of cancer screening modalities. MATERIALS AND METHODS: During the early recruitment phase of PLCO (1993-1997), data on recruitment outcome were collected at the Henry Ford Health System (HFHS) in Detroit, Michigan. In this phase, HFHS identified potential participants using patient databases. Records were used to assess recruitment success by age, sex, race, household income (using area-based U.S. Census data), and preexisting morbidity. Logistic regression was used to assess whether enrollment success differed significantly according to these factors. RESULTS: Of 74,139 persons ages 55 to 74 invited by HFHS to participate, 8,250 (11%) ;enrolled. In multivariate analyses, the odds of enrolling were modestly but significantly higher for women, Caucasians, persons in their 60's, and persons living in census blocks with higher median household income. Persons with two or more preexisting morbidities had significantly lower odds of enrolling compared to those with one or no preexisting morbidities. CONCLUSIONS: These data suggest that only a small fraction of persons invited to enroll in long-term RCTs of cancer screening modalities actually do so. In this urban, Midwestern setting, certain characteristics including age, race, and income influenced recruitment success, albeit modestly. PMID- 18398024 TI - Health insurance-related disparities in colorectal cancer screening in Virginia. AB - BACKGROUND: Colorectal cancer (CRC) screening rates remain low in the United States. The effect of health insurance on CRC screening is not clear. We assessed the association between having health insurance and being screened for CRC and the factors that modify this association. METHODS: We used data from the 2005 Virginia Behavioral Risk Factor Surveillance System to evaluate the association of self-reported insurance coverage on self-reported CRC screening among all men and women ages > or =50 years (N = 2,887). Prevalence odds ratios (POR) were estimated using unconditional logistic regression. All covariates were assessed for potential effect measure modification and confounding. All analyses accounted for the Behavioral Risk Factor Surveillance System complex survey sampling design. RESULTS: Overall, participants who reported having insurance coverage were more than twice as likely to report being screened for CRC compared with those who reported having none [crude POR, 2.16; 95% confidence interval (95% CI), 1.26-3.68]. This relationship differed between men and women (POR(males), 3.37; 95% CI, 1.63-6.96; POR(females), 1.46; 95% CI, 0.74-2.89). After adjusting for age and income, self-reported insurance coverage had a positive association with report of being screened among men (POR, 2.02; 95% CI, 0.96-4.23) but not among women (POR, 0.81; 95% CI, 0.34-1.93). CONCLUSIONS: Men who reported having health insurance were more likely to report having CRC screening than those who reported not having insurance coverage. However, this effect was not observed in women. These findings, if confirmed in other study populations, indicate that improving CRC screening coverage may require not only insurance status specifications but also gender-explicit considerations. PMID- 18398025 TI - Cancer high-risk subjects identified by serum pepsinogen tests: outcomes after 10 year follow-up in asymptomatic middle-aged males. AB - BACKGROUND: Gastric cancer screening using the pepsinogen filter test is receiving wide recognition in Japan owing to convenience, freedom from discomfort or risk, efficiency, and economy. Because the long-term outcomes of cancer development in extensive atrophic gastritis detected by pepsinogen test are unclear, test-positive and test-negative subjects were investigated in a longitudinal cohort study. METHODS: Subjects comprised 5,209 middle-aged men with measured serum pepsinogen levels who were followed for 10 years. Cancer development based on "atrophy-positive" and "atrophy-negative" criteria used for cancer screening was investigated. RESULTS: During the study, 63 cases of cancer developed in the cohort, representing an incidence rate of 125 per 100,000 person years. Pepsinogen test screening using the most widely used atrophy-positive criterion (pepsinogen I, < or =70 ng/mL; pepsinogen I/II ratio, < or =3.0) displayed 58.7% sensitivity, 73.4% specificity, and 2.6% positive predictive value. Cancer incidence rate was 276 per 100,000 person-years for the atrophy positive group and 70 per 100,000 person-years for the atrophy-negative group. Incidence rate was higher in groups fulfilling stricter positive criteria detecting more extensive atrophy, reaching 424 per 100,000 person-years. In addition, 9.2% of atrophy-negative subjects with pepsinogen I of >70 ng/mL and pepsinogen I/II ratio of < or =3.0 (reflecting putative inflammation-based high pepsinogen II level) are at high risk for cancer, particularly diffuse-type cancer, with a cancer incidence rate comparable with atrophy-positive subjects (216 per 100,000 person-years). CONCLUSION: Atrophy-positive subjects by pepsinogen filter test, particularly those fulfilling stricter criteria, and atrophy-negative subjects with low pepsinogen I/II ratio reflecting putative extensive active inflammation constitute populations at high risk for gastric cancer, requiring thorough endoscopic examination. PMID- 18398026 TI - Risk factors for hepatocellular carcinoma in a Japanese population: a nested case control study. AB - BACKGROUND: Epidemiologic studies have shown effects of lifestyle-related factors on risk for hepatocellular carcinoma. However, few cohort studies have incorporated, in a strict and in-depth manner, hepatitis B virus (HBV) and hepatitis C virus (HCV) infections or investigated synergism between such factors. METHODS: We conducted a nested case-control study using sera stored before hepatocellular carcinoma diagnosis in the longitudinal cohort of atomic bomb survivors. The study included 224 hepatocellular carcinoma cases and 644 controls that were matched to the cases on gender, age, city, time of serum storage, and method of serum storage, and countermatched on radiation dose. RESULTS: Univariate analysis showed that HBV and HCV infections, alcohol consumption, smoking habit, body mass index (BMI), and diabetes mellitus were associated with increased hepatocellular carcinoma risk, whereas coffee drinking was associated with decreased hepatocellular carcinoma risk. Multivariate relative risks of hepatocellular carcinoma (95% confidence interval) were 45.8 (15.2-138), 101 (38.7-263), 70.7 (8.3-601), 4.36 (1.48-13.0), and 4.57 (1.85 11.3), for HBV infection alone, HCV infection alone, both HBV and HCV infections, alcohol consumption of > or =40 g of ethanol per day, and BMI of >25.0 kg/m(2) 10 years before diagnosis, respectively. HBV and HCV infection and BMI of >25.0 kg/m(2) remained independent risk factors even after adjusting for severity of liver fibrosis. Among HCV-infected individuals, the relative risk of hepatocellular carcinoma for a 1 kg/m(2) increase in BMI was 1.39 (P = 0.003). CONCLUSIONS: To limit the risk for hepatocellular carcinoma, control of excess weight may be crucial for individuals with chronic liver disease, especially those with chronic hepatitis C. PMID- 18398027 TI - Impact of estrogen deprivation on gene expression profiles of normal postmenopausal breast tissue in vivo. AB - Aromatase inhibitors play a key role in the clinical management of hormone receptor-positive breast cancer and have potential utility as chemopreventive agents. Further understanding of the molecular effects of estrogen and its deprivation in normal breast tissue may allow the development of biomarkers of risk of breast cancer and help to predict the value of chemoprevention with aromatase inhibitors. Core biopsies of normal breast tissue were taken before and after letrozole treatment from postmenopausal women in the LITMaS pilot prevention study. RNA was extracted from these samples and used for cDNA microarray analysis. Gene expression changes induced by letrozole treatment were much less extensive than observed in estrogen receptor-positive malignant tissue; however, overall, they correlated to a highly significant degree (rho = 0.511; P < 10(-20)). As well as some classically estrogen-associated genes, many genes associated with extracellular matrix remodeling were affected by estrogen deprivation in the normal breast in vivo. These data indicate for the first time that gene expression of normal breast tissue remains dependent on endogenous estrogens after the menopause. The modest degree of gene change suggests that intermediate markers of chemoprevention may be difficult to identify. PMID- 18398028 TI - Association of mammographic density with the pathology of subsequent breast cancer among postmenopausal women. AB - BACKGROUND: Limited studies have examined the associations between mammographic density and subsequent breast tumor characteristics. METHODS: Eligible women were part of a case-control study of postmenopausal breast cancer, were 40 years or older and had a routine mammogram 4 years or more before their diagnosis. Mammographic density (percent density, dense area, and nondense area) was estimated using a computer-assisted thresholding program. At the time of cancer diagnosis, cases were classified as asymptomatic or symptomatic based on medical record review and breast imaging workup. Pathologic review was done blinded to the density status. Linear regression models and tests for trend examined the association between pathologic characteristics of the breast tumor and the components of density for all participants, and stratified by symptom status at diagnosis. RESULTS: Of the 286 eligible cases, 77% were 60 years or older and mean percent density was 29.5% (SD, 14.6%). Density was not significantly associated with tumor size (P = 0.22), histologic type (P = 0.77), estrogen receptor (P = 0.11) or progesterone receptor (P = 0.37) status, mitotic activity (P = 0.12), or nuclear pleomorphism (P = 0.09; P values for percent density). An inverse association was suggested between tumor grade and percent density (32.0%, 30.3%, 26.7% for grades 1-3; P = 0.06 for trend). The inverse association with tumor grade and its components (nuclear pleomorphism and tubular differentiation) was only evident among the 97 symptomatic women; positive associations of estrogen receptor (P = 0.009) and progesterone receptor (P = 0.04) were also seen with percent density only in this subgroup. CONCLUSIONS: The inverse association between tumor grade and percent density in the symptomatic population could inform the biology of the association between mammographic density and breast cancer risk. PMID- 18398029 TI - Genetic polymorphisms involved in insulin-like growth factor (IGF) pathway in relation to mammographic breast density and IGF levels. AB - The insulin-like growth factor (IGF) pathway is believed to play a role in carcinogenesis of the mammary gland. Single nucleotide polymorphisms (SNPs) of IGF-I, IGF-binding protein-3 (IGFBP-3), IGF receptor 1, insulin receptor substrate 1, and phosphoinositide-3-kinase, catalytic, beta polypeptide genes, which are members of the IGF pathway, have been associated with risk of common cancers, breast density, and/or IGF levels but results remain inconclusive. Thus, we evaluated the association of 11 targeted IGF pathway SNPs with circulating IGF levels and mammographic breast density. Among 741 white premenopausal women, blood samples were collected at time of screening mammography, and plasma IGF-I and IGFBP-3 levels were measured by ELISA. Percent and absolute breast density were estimated using a computer-assisted method. Multivariate linear models were used to examine the associations. Women carrying increasing number of copies of the rare allele of IGF-I rs1520220 and rs6220 SNPs had increased percent breast density (P(trend) = 0.04 and 0.06, respectively). Carriers of increasing number of copies of the rare allele of phosphoinositide-3-kinase, catalytic, beta polypeptide rs361072 SNP had decreased percent (P(trend) = 0.04) and absolute (P(trend) = 0.02) breast density. An association of insulin receptor substrate 1 rs1801278 SNP with absolute density (P(trend) = 0.03) was also observed. All four IGFBP-3 SNPs (including rs2854744) were associated with IGF-I and IGFBP-3 levels. This study shows that several components of the IGF pathway are associated with breast density or IGF levels. Our findings provide additional support for the idea that several components of the IGF pathway may affect breast cancer risk and that this effect on breast cancer development may be mediated, at least in part, through its influence on the morphogenesis of breast tissue. PMID- 18398030 TI - Coffee consumption and CYP1A2*1F genotype modify age at breast cancer diagnosis and estrogen receptor status. AB - CYP1A2 plays a key role in the metabolism of both estrogen and coffee. Women with higher coffee intake and the CYP1A2*1F A/A genotype have a ratio of high 2 hydroxyestrone (2-OHE1) to 16alpha-OHE1. 2-OHE1 is a weak estrogen and may even block the estrogen receptor (ER), whereas 16alpha-OHE1 is procarcinogenic. We hypothesized that moderate to high coffee consumption (> or =2 cups per day) combined with the CYP1A2*1F A/A genotype would be associated with a later age at diagnosis and a greater proportion of ER-negative (ER-) tumors among patients with breast cancer. We genotyped 458 patients with breast cancer (age, 25-99 years) in Lund, Sweden, for CYP1A2*1F. Information on lifestyle factors and tumor characteristics were obtained from preoperative questionnaires and pathology reports. Among patients with CYP1A2*1F A/A (51.3%), moderate to high consumption was associated with a later age at diagnosis compared with low coffee consumption (59.8 versus 52.6 years, P = 0.0004). These patients were also more likely to have ER- tumors than patients with low consumption (14.7% versus 0%, P = 0.018). Coffee was not associated with ER status or age at diagnosis in patients with at least one C allele. Age at diagnosis was not associated with ER status in patients with CYP1A2*1F A/A, but younger patients (<50 years) with at least one C allele were more likely to have ER- tumors compared with older patients (odds ratio, 4.2; 95% confidence interval, 1.9-9.3; P = 0.0002). These findings raise the hypothesis that coffee slows the growth of ER-positive tumors in patients with CYP1A2*1F A/A and may have implications for breast cancer if confirmed. PMID- 18398031 TI - BTF4/BTNA3.2 and GCS as candidate mRNA prognostic markers in epithelial ovarian cancer. AB - This study aims to identify reliable prognosis markers to predict patient outcome at surgery in high-grade serous epithelial ovarian cancer by a real-time quantitative PCR (RT-q-PCR)-based test. Seventeen tissue samples from serous epithelial ovarian cancer patients were screened by DNA microarray to identify genes differentially expressed between tumors from patients who relapsed within 18 months and tumors from patients showing no relapse or relapsed after 24 months after initial diagnosis. RNA expression of a subset of genes was validated by RT q-PCR in the initial set of 17 samples. From these results, a refined list was selected and tested in independent samples from 41 serous. Expression was associated with time to relapse and clinical variables. Microarray analysis identified a profile of 34 differentially expressed genes. RT-q-PCR validated the expression profile of a subset of seven genes in the initial set of patients. Differential gene expression was also validated in an independent set of patients. Low BTF4 or GCS expression was strongly associated with poor outcome in Kaplan-Meier analysis (P < 0.05, log-rank test) and Cox univariate as well as in multivariate analyses with a higher hazard ratio than clinical variables, such as residual disease, age, stage, and grade. PMID- 18398032 TI - A prospective evaluation of insulin and insulin-like growth factor-I as risk factors for endometrial cancer. AB - Obesity is a major risk factor for endometrial cancer, a relationship thought to be largely explained by the prevalence of high estrogen levels in obese women. Obesity is also associated with high levels of insulin, a known mitogen. However, no prospective studies have directly assessed whether insulin and/or insulin-like growth factor-I (IGF-I), a related hormone, are associated with endometrial cancer while accounting for estrogen levels. We therefore conducted a case-cohort study of incident endometrial cancer in the Women's Health Initiative Observational Study, a prospective cohort of 93,676 postmenopausal women. The study involved all 250 incident cases and a random subcohort of 465 subjects for comparison. Insulin, total IGF-I, free IGF-I, IGF-binding protein-3, glucose, and estradiol levels were measured in fasting baseline serum specimens. Cox models were used to estimate associations with endometrial cancer, particularly endometrioid adenocarcinomas, the main histologic type (n = 205). Our data showed that insulin levels were positively associated with endometrioid adenocarcinoma [hazard ratio contrasting highest versus lowest quartile (HR(q4-q1)), 2.33; 95% confidence interval (95% CI), 1.13-4.82] among women not using hormone therapy after adjustment for age and estradiol. Free IGF-I was inversely associated with endometrioid adenocarcinoma (HR(q4-q1), 0.53; 95% CI, 0.31-0.90) after adjustment for age, hormone therapy use, and estradiol. Both of these associations were stronger among overweight/obese women, especially the association between insulin and endometrioid adenocarcinoma (HR(q4-q1), 4.30; 95% CI, 1.62-11.43). These data indicate that hyperinsulinemia may represent a risk factor for endometrioid adenocarcinoma that is independent of estradiol. Free IGF-I levels were inversely associated with endometrioid adenocarcinoma, consistent with prior cross sectional data. PMID- 18398034 TI - Consumption of raw cruciferous vegetables is inversely associated with bladder cancer risk. AB - Cruciferous vegetables contain isothiocyanates, which show potent chemopreventive activity against bladder cancer in both in vitro and in vivo studies. However, previous epidemiologic studies investigating cruciferous vegetable intake and bladder cancer risk have been inconsistent. Cooking can substantially reduce or destroy isothiocyanates, and could account for study inconsistencies. In this hospital-based case-control study involving 275 individuals with incident, primary bladder cancer and 825 individuals without cancer, we examined the usual prediagnostic intake of raw and cooked cruciferous vegetables in relation to bladder cancer risk. Odds ratios (OR) and 95% confidence intervals (CI) were estimated with unconditional logistic regression, adjusting for smoking and other bladder cancer risk factors. We observed a strong and statistically significant inverse association between bladder cancer risk and raw cruciferous vegetable intake (adjusted OR for highest versus lowest category = 0.64; 95% CI, 0.42 0.97), with a significant trend (P = 0.003); there were no significant associations for fruit, total vegetables, or total cruciferous vegetables. The associations observed for total raw crucifers were also observed for individual raw crucifers. The inverse association remained significant among current and heavy smokers with three or more servings per month of raw cruciferous vegetables (adjusted ORs, 0.46 and 0.60; 95% CI, 0.23-0.93 and 0.38-0.93, respectively). These data suggest that cruciferous vegetables, when consumed raw, may reduce the risk of bladder cancer, an effect consistent with the role of dietary isothiocyanates as chemopreventive agents against bladder cancer. PMID- 18398033 TI - Dairy product, saturated fatty acid, and calcium intake and prostate cancer in a prospective cohort of Japanese men. AB - Many epidemiologic studies have reported a positive association between dairy products and prostate cancer. Calcium or saturated fatty acid in dairy products has been suspected as the causative agent. To investigate the association between dairy products, calcium, and saturated fatty acid and prostate cancer in Japan, where both the intake of these items and the incidence of prostate cancer are low, we conducted a population-based prospective study in 43,435 Japanese men ages 45 to 74 years. Participants responded to a validated questionnaire that included 138 food items. During 7.5 years of follow-up, 329 men were newly diagnosed with prostate cancer. Dairy products were associated with a dose dependent increase in the risk of prostate cancer. The relative risks (95% confidence intervals) comparing the highest with the lowest quartiles of total dairy products, milk, and yogurt were 1.63 (1.14-2.32), 1.53 (1.07-2.19), and 1.52 (1.10-2.12), respectively. A statistically significant increase in risk was observed for both calcium and saturated fatty acid, but the associations for these were attenuated after controlling for potential confounding factors. Some specific saturated fatty acids increased the risk of prostate cancer in a dose dependent manner. Relative risks (95% confidence intervals) on comparison of the highest with the lowest quartiles of myristic acid and palmitic acid were 1.62 (1.15-2.29) and 1.53 (1.07-2.20), respectively. In conclusion, our results suggest that the intake of dairy products may be associated with an increased risk of prostate cancer. PMID- 18398035 TI - Detection and quantitation of N'-nitrosonornicotine in human toenails by liquid chromatography-electrospray ionization-tandem mass spectrometry. AB - Specific biomarkers of tobacco carcinogen uptake are critical for investigations of the role of tobacco smoke exposure in human cancers. Two new biomarkers of human exposure to tobacco-specific carcinogens have been recently developed by our research group: urinary N'-nitrosonornicotine (NNN) and toenail 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). In this study, we report the presence of NNN in human toenails. Toenails of 17 smokers were analyzed for total NNN. Mean total NNN level in these samples was 4.63 +/- 6.48 fmol/mg toenail and correlated with previously reported total NNAL (r = 0.96; P < 0.0001), total nicotine (r = 0.48; P < 0.05), and total cotinine (r = 0.87; P < 0.0001). An interesting finding was that amounts of NNN in smokers' toenails were generally higher than those of total NNAL. The ratio of toenail NNN to NNAL averaged 2.8, whereas the previously reported ratio between these biomarkers in smokers' urine was 0.1. NNN was also found in toenail samples from 12 nonsmokers, averaging 0.35 +/- 0.16 fmol/mg and positively correlating with toenail cotinine (r = 0.58; P = 0.05). The results of this study show the feasibility of quantifying NNN in human toenails, providing a potentially useful new biomarker of tobacco carcinogen exposure. PMID- 18398036 TI - Prognostic significance of cadherin-based adhesion molecules in cutaneous malignant melanoma. AB - BACKGROUND: The need for novel molecular prognostic markers that can supplement validated clinicopathologic correlates for cutaneous malignant melanoma is well recognized. Proteins that mediate the epithelial-mesenchymal transition, the process by which a cancer cell disengages from its parent tumor, are important candidates. METHODS: The prognostic relevance of E-cadherin, N-cadherin, and P cadherin, calcium-dependent transmembrane glycoproteins that regulate cell-cell adhesion, and their adaptors, alpha-catenin, beta-catenin, and p120-catenin, was evaluated on a cohort of 201 primary and 274 metastatic melanoma tumors using fluorescence-based immunohistochemical methods and Automated Quantitative Analysis of protein expression on digitally captured photomicrographs. RESULTS: Increasing levels of N-cadherin expression improved overall survival (log-rank = 7.31; P = 0.03) but did not retain significance following adjustment for established clinicopathologic correlates (P = 0.50). Higher levels of E-cadherin approached significance for favorable prognosis on both univariate (P = 0.13) and multivariable (P = 0.10) analyses. Hierarchical clustering of the composite profiles for all six markers identified four unique clusters that yielded differential overall survival (log-rank = 10.54; P = 0.01). Cluster 4, expressing high E-cadherin and N-cadherin levels, possessed the most favorable outcome and cluster 2, featuring low E-cadherin and alpha-catenin but modest N-cadherin, showed least favorable outcomes. Cluster 2 remained significant on multivariable analysis (hazard ratio, 3.29; 95% confidence interval, 1.50-7.19; P = 0.003). CONCLUSIONS: Although none of the cadherin-based adhesion molecules were independently prognostic, multimarker profiles were significant. Similar to epithelial-derived tumors, loss of E-cadherin correlates with poor outcome. In contrast, for neural crest-derived cutaneous malignant melanoma, N-cadherin overexpression can be associated with either a successful epithelial-mesenchymal transition or a favorably differentiated tumor. Additional cadherin profiles are needed to discriminate these distinctive phenotypes. PMID- 18398037 TI - Family history of cancer and associated risk of developing neuroendocrine tumors: a case-control study. AB - BACKGROUND: Carcinoids are rare neuroendocrine tumors (NET). Familial clusterings of NETs are rarely reported, except for a small proportion associated with multiple endocrine neoplasia syndrome type 1. We evaluated the effect of positive family history of NET as well as other cancers on the development of NETs arising at five different locations. METHODS: We conducted a retrospective, hospital based, case-control study involving 740 patients with histologically confirmed NETs and 924 healthy controls. Information on family history of cancer was collected, and unconditional logistic regression analysis was used to determine adjusted odds ratios (AOR) and 95% confidence intervals (CI). RESULTS: The authors observed a significant relationship between first-degree relatives with cancers and the development of NETs arising at the small intestine, stomach, lung, and pancreas; AORs (95% CI) were 1.6 (1.1-2.4), 2.5 (1.1-6.3), 2.6 (1.5 4.5), and 1.8 (1.1-3.1), respectively. A first-degree family history of esophageal cancer was significantly associated with pancreatic NETs; AOR, 5.6 (95% CI, 1.1-29.6). There was a 70% and 130% increased risk of developing small intestinal NETs among subjects with family histories of colorectal cancer and prostate cancer, respectively. Moreover, individuals with a family history of lung cancer had a 2-fold increase in risk of developing pulmonary NETs. CONCLUSIONS: Having a first-degree relative with any cancer in general, and NET in particular, was a risk factor for NETs. The elevated risk of developing NETs extended to individuals with a family history of other cancers (not NETs) among first-degree relatives. These results suggested that risk of NETs may be partially explained by genetic factors. PMID- 18398038 TI - Line region hypomethylation is associated with lifestyle and differs by human papillomavirus status in head and neck squamous cell carcinomas. AB - Genomic hypomethylation is a hallmark of essentially all cancers, but the degree of this hypomethylation differs among individual tumors. Little work has explored what leads to these differences and or asked whether they are clinically meaningful. In this study of head and neck squamous cell carcinoma, we assessed hypomethylation in tumors using a semiquantitative fragment analysis approach to determine the relative methylation status of the line retroviral element LRE1 (Line-1.2). Because this is an established marker of genomic methylation status, we examined the relationship between the relative methylation, patient demographics, and other risk factors for head and neck squamous cell carcinoma. We determined relative methylation status for 303 patients, 193 of which had complete data for all variables of interest. Using a generalized linear model, we found that patient body mass index was significantly positively associated with tumor LRE1 methylation level. Smoking duration, particularly in tumors lacking human papillomavirus (HPV) DNA, was significantly negatively associated with relative methylation level. Having previously assessed relative methylation in blood-derived DNA, we compared tumor with the blood DNA methylation level and observed these to be independent. Finally, the lower LRE1 methylation in patients whose tumors were HPV DNA negative was associated with poorer patient survival (hazard ratio, 1.6; 95% confidence interval, 1.0-2.6). These findings suggest that HPV-associated tumors differ molecularly from those arising after heavy tobacco use and that this epigenetic alteration may affect survival in HPV negative patients already exhibiting a more aggressive disease. PMID- 18398039 TI - Polymorphisms in angiogenesis-related genes and prostate cancer. AB - BACKGROUND: Angiogenesis is required for development and progression of prostate cancer. Potentially functional single nucleotide polymorphisms (SNP) in genes important in prostate angiogenesis (VEGF, HIF1A, and NOS3) have previously been associated with risk or severity of prostate cancer. METHODS: Prostate cancer cases (n = 1,425) and controls (n = 1,453) were selected from the Cancer Prevention Study II Nutrition Cohort. We examined associations between 58 SNPs in nine angiogenesis-related candidate genes (EGF, LTA, HIF1A, HIF1AN, MMP2, MMP9, NOS2A, NOS3, VEGF) and risk of overall and advanced prostate cancer. Unconditional logistic regression was used to estimate odds ratios, adjusted for matching factors. RESULTS: Our results did not replicate previously observed associations with SNPs in VEGF, HIF1A, or NOS3, nor did we observe associations with SNPs in EGF, LTA, HIF1AN, MMP9, or NOS2A. In the MMP2 gene, three intronic SNPs, all in linkage disequilibrium, were associated with overall and advanced prostate cancer (for overall prostate cancer, P(trend) = 0.01 for rs1477017, P(trend) = 0.01 for rs17301608, P(trend) = 0.02 for rs11639960). However, two of these SNPs (rs17301608 and rs11639960) were examined and were not associated with prostate cancer in a recent genome-wide association study using prostate cancer cases and controls from the Prostate, Lung, Colorectal, and Ovary study cohort. Furthermore, when we pooled our results for these two SNPs with those from the Prostate, Lung, Colorectal, and Ovary cohort; neither SNP was associated with prostate cancer. CONCLUSION: None of the SNPs examined seem likely to be importantly associated with risk of overall or advanced prostate cancer. PMID- 18398041 TI - MDM2 SNP309 is associated with endometrial cancer risk. AB - Mouse double-minute 2 homologue (MDM2) is a key negative regulator of p53, a tumor suppressor gene that initiates cell cycle arrest and apoptosis in response to DNA damage and other cellular stresses. A T > G polymorphism found in the promoter region of MDM2 (SNP309) increases MDM2 expression and thereby attenuates p53 activity. We genotyped the MDM2 polymorphism SNP309 in endometrial cancer case-control studies nested within the Nurses' Health Study (454 cases and 1,132 controls) and the Women's Health Study (137 cases and 411 controls). Due to a significant difference in genotype distribution by ethnicity, we restricted our analyses to Caucasians. We calculated odds ratios and 95% confidence intervals using conditional and unconditional logistic regression adjusted for age at menarche, parity and age at first birth, postmenopausal hormone use at diagnosis, age at menopause and menopausal status at diagnosis, first-degree family history of colon cancer, body mass index at diagnosis, and cigarette smoking status at diagnosis. Women with a heterozygous genotype had no greater risk whereas those with a homozygous variant genotype had a greater risk than women with a wild-type genotype for the MDM2 SNP309 (covariate-adjusted odds ratio, 1.87; 95% confidence interval, 1.29-2.73) for endometrial cancer. We observed no association between age at diagnosis and genotype. Women carrying two copies of the MDM2 SNP309 variant may be at greater risk of endometrial cancer. PMID- 18398040 TI - Transcription factor 7-like 2 polymorphism and colon cancer. AB - Polymorphisms of the transcription factor 7-like 2 (TCF7L2) gene have been associated with insulin sensitivity and diabetes, and the TCF7L2 gene is involved in the Wnt/beta-catenin signaling pathway, all factors thought to be important in the etiology of colon cancer. In this confirmatory study, we evaluated the rs7903146 TCF7L2 polymorphism with colon cancer using previously collected data on 1,578 cases and 1,966 controls. We did not observe a statistically significant association between the rs7903146 polymorphisms and risk of colon cancer [odds ratio (OR), 1.12; 95% confidence interval (95% CI), 0.98-1.28] when evaluating the total population. We did, however, observe a statistically significant interaction between the rs7903146 TCF7L2 polymorphism and recent use of aspirin/nonsteroidal anti-inflammatory drugs (NSAID; P = 0.001). Increased colon cancer risk associated with the T allele was restricted to those without recent use of aspirin/NSAIDs (OR, 1.65; 95% CI, 1.35-2.02, relative to recent aspirin users, i.e., use of aspirin/NSAIDS within the 2 years before diagnosis, with the CC genotype). Among individuals who reported recent use of aspirin/NSAIDs, the T allele reduced risk of colon cancer (OR, 0.78; 95% CI, 0.62-0.98) in a dose response fashion (P for linear trend across genotypes = 0.03). These data suggest that colon cancer risk associated with the rs7903146 TCF7L2 polymorphism is modified by use of aspirin/NSAIDs. PMID- 18398042 TI - The common D302H variant of CASP8 is associated with risk of glioma. AB - Caspase 8 (CASP8) is a key regulator of apoptosis or programmed cell death, and, hence, a defense against cancer. We tested the hypothesis that the CASP8 polymorphism D302H influences risk of glioma through analysis of five series of glioma case patients and controls (n = 1,005 and 1,011, respectively). Carrier status for the rare allele of D302H was associated with a 1.37-fold increased risk (95% confidence interval, 1.10-1.70; P = 0.004). The association of CASP8 D302H with glioma risk indicates the importance of inherited variation in the apoptosis pathway in susceptibility to this form of primary brain tumor. PMID- 18398043 TI - No evidence for differences in DNA damage assessed before and after a cancer diagnosis. AB - The overwhelming majority of studies that have found increased cancer risk associated with functional deficits in DNA repair used a case-control design, in which measurements were made after cancer diagnosis. However, there are concerns about whether the cancer itself or cancer treatment affected the conclusions (reverse causation bias). We assessed the effect of cancer diagnosis among 26 breast cancer controls who had blood collected during 2001 to 2003 and again in 2005 to 2006 after being diagnosed with cancer. Using the alkaline comet assay, we quantified DNA damage in untreated lymphoblastoid cell lines. Comet distributed moment, olive tail moment, percentage of DNA in tail, and comet tail length were summarized as the geometric mean of 100 cells. For comet distributed moment, olive tail moment, tail DNA, and tail length, the proportions of women with before diagnosis values higher than after diagnosis were 65%, 50%, 50%, and 46%, respectively. We found no significant differences in the before or after diagnosis mean comet values. Median cut-points were determined from the before diagnosis distribution, and we used conditional logistic regression to calculate odds ratios (OR) and upper 95% bounds of the confidence intervals. ORs ranged from 0.6 to 0.9 with upper confidence interval bounds of 1.9 and 2.6, meaning biased ORs above 2.6 are unlikely. We found no evidence that reverse causation bias is an important concern in case-control studies using the comet assay applied to cell lines collected after cancer diagnosis. More work is needed to characterize the effect of cancer diagnosis on other phenotypic assays. PMID- 18398044 TI - Differential methylation of a short CpG-rich sequence within exon 1 of TCF21 gene: a promising cancer biomarker assay. AB - Detection of cancer cells at early stages could potentially increase survival rates in cancer patients. Aberrant promoter hypermethylation is a major mechanism for silencing tumor suppressor genes in many kinds of human cancers. A recent report from our laboratory described the use of quantitative methylation-specific PCR assays for discriminating patients with lung cancer from those without lung cancer using lung biopsies as well as sputum samples. TCF21 is known to be essential for differentiation of epithelial cells adjacent to mesenchyme. Using restriction landmark genomic scanning, a recent study identified TCF21 as candidate tumor suppressor at 6q23-q24 that is epigenetically inactivated in lung and head and neck cancers. Using DNA sequencing technique, we narrowed down a short CpG-rich segment (eight specific CpG sites in the CpG island within exon 1) of the TCF21 gene, which was unmethylated in normal lung epithelial cells but predominantly methylated in lung cancer cell lines. We specifically targeted this short CpG-rich sequence and developed a quantitative methylation-specific PCR assay suitable for high-throughput analysis. We showed the usefulness of this assay in discriminating patients with lung cancer from those without lung cancer using biopsies and sputum samples. We further showed similar applications with multiple other malignancies. Our assay might have important implications in early detection and surveillance of multiple malignancies. PMID- 18398045 TI - Association between genetic polymorphisms of macrophage scavenger receptor 1 gene and risk of prostate cancer in the health professionals follow-up study. AB - BACKGROUND: Macrophage scavenger receptor 1 (MSR1) is involved in chronic inflammation, which is a risk factor for prostate cancer. Association studies assessing the relationship between sequence variants of MSR1 and prostate cancer are inconsistent. We hypothesized that sequence variants of MSR1 were associated with prostate cancer risk. METHODS: In a nested case-control design within the Health Professionals Follow-up Study, we identified 700 participants with prostate cancer diagnosed after they had provided a blood specimen in 1993 and before January 2000. Controls were 700 age-matched men without prostate cancer who had had a prostate-specific antigen test after providing a blood specimen. We genotyped three common (>5%) single nucleotide polymorphisms (SNP) that have been reported previously to be associated with risk of prostate cancer. RESULTS: None of these MSR1 SNPs nor estimated haplotypes were associated with prostate cancer risk (P for the global test for haplotypes = 0.89). These MSR1 SNPs also did not appear to be associated with higher-grade or advanced-stage prostate cancer. CONCLUSION: The association between these sequence variants of MSR1 and the risk of prostate cancer was null. Further study of aggressive prostate cancer may be warranted, as we had limited power to assess these. PMID- 18398046 TI - Lack of association between physical activity in smokers and plasma glutathione peroxidase levels. PMID- 18398047 TI - Single nucleotide polymorphisms in obesity-related genes and the risk of esophageal cancers. AB - Rates of adenocarcinoma of the esophagus (EAC) and esophagogastric junction (EGJAC) have been rising rapidly in recent decades, in contrast to the declining rates of esophageal squamous cell carcinomas (ESCC). Obesity is a major risk factor for both EAC and EGJAC, but not ESCC, and there is speculation that obesity promotes adenocarcinoma development through endocrine and related pathways. We therefore compared the prevalence of 12 single nucleotide polymorphisms (SNPs) in nine candidate genes previously implicated in obesity pathways (LEP, LEPR, ADIPOQ, POMC, PPARalpha, PPARgamma, RXRgamma, GHRL, and INSIG2) in a large Australian case-control study comprising DNA samples from 260 EAC cases, 301 EGJAC cases, 213 ESCC cases, and 1,352 population controls. No SNPs were associated with EGJAC or ESCC. Although several SNPs seemed to be associated with EAC on crude analysis [ADIPOQ (rs1501299), LEP (5'-untranslated region), PPARgamma (H447H), and GHRL (M72L)], effect sizes were modest and none of the associations was significant after correcting for multiple comparisons. Further, we found no consistent evidence that any of the genotypes were associated with risk of EAC or EGJAC within strata of body mass index (<25.0 kg/m(2), 25.0-29.9 kg/m(2), >30 kg/m(2)). In conclusion, our data suggest that these SNPs do not play a major role in esophageal carcinogenesis. PMID- 18398048 TI - The ratio of specific polychlorinated biphenyls as a surrogate biomarker of cytochrome P4501A2 activity: a pharmaco-metabonomic study in humans. PMID- 18398049 TI - Polymorphisms in RAD51, XRCC2, and XRCC3 are not related to breast cancer risk. PMID- 18398050 TI - Histone acetylation and chromatin remodeling are required for UV-B-dependent transcriptional activation of regulated genes in maize. AB - The nuclear proteomes of maize (Zea mays) lines that differ in UV-B tolerance were compared by two-dimensional gel electrophoresis after UV light treatment. Differential accumulation of chromatin proteins, particularly histones, constituted the largest class identified by mass spectrometry. UV-B-tolerant landraces and the B73 inbred line show twice as many protein changes as the UV-B sensitive b, pl W23 inbred line and transgenic maize expressing RNA interference constructs directed against chromatin factors. Mass spectrometic analysis of posttranslational modifications on histone proteins demonstrates that UV-B tolerant lines exhibit greater acetylation on N-terminal tails of histones H3 and H4 after irradiation. These acetylated histones are enriched in the promoter and transcribed regions of the two UV-B-upregulated genes examined; radiation sensitive lines lack this enrichment. DNase I and micrococcal nuclease hypersensitivity assays indicate that chromatin adopts looser structures around the selected genes in the UV-B-tolerant samples. Chromatin immunoprecipitation experiments identified additional chromatin factor changes associated with the nfc102 test gene after UV-B treatment in radiation-tolerant lines. Chromatin remodeling is thus shown to be a key process in acclimation to UV-B, and lines deficient in this process are more sensitive to UV-B. PMID- 18398051 TI - Thylakoid membrane remodeling during state transitions in Arabidopsis. AB - Adaptability of oxygenic photosynthetic organisms to fluctuations in light spectral composition and intensity is conferred by state transitions, short-term regulatory processes that enable the photosynthetic apparatus to rapidly adjust to variations in light quality. In green algae and higher plants, these processes are accompanied by reversible structural rearrangements in the thylakoid membranes. We studied these structural changes in the thylakoid membranes of Arabidopsis thaliana chloroplasts using atomic force microscopy, scanning and transmission electron microscopy, and confocal imaging. Based on our results and on the recently determined three-dimensional structure of higher-plant thylakoids trapped in one of the two major light-adapted states, we propose a model for the transitions in membrane architecture. The model suggests that reorganization of the membranes involves fission and fusion events that occur at the interface between the appressed (granal) and nonappressed (stroma lamellar) domains of the thylakoid membranes. Vertical and lateral displacements of the grana layers presumably follow these localized events, eventually leading to macroscopic rearrangements of the entire membrane network. PMID- 18398052 TI - Identification and regulation of TPS04/GES, an Arabidopsis geranyllinalool synthase catalyzing the first step in the formation of the insect-induced volatile C16-homoterpene TMTT. AB - Volatile secondary metabolites emitted by plants contribute to plant-plant, plant fungus, and plant-insect interactions. The C(16)-homoterpene TMTT (for 4,8,12 trimethyltrideca-1,3,7,11-tetraene) is emitted after herbivore attack by a wide variety of plant species, including Arabidopsis thaliana, and is assumed to play a role in attracting predators or parasitoids of herbivores. TMTT has been suggested to be formed as a degradation product of the diterpene alcohol (E,E) geranyllinalool. Here, we report the identification of Terpene Synthase 04 (TPS04; At1g61120) as a geranyllinalool synthase (GES). Recombinant TPS04/GES protein expressed in Escherichia coli catalyzes the formation of (E,E) geranyllinalool from the substrate geranylgeranyl diphosphate. Transgenic Arabidopsis lines carrying T-DNA insertions in the TPS04 locus are deficient in (E,E)-geranyllinalool and TMTT synthesis, a phenotype that can be complemented by expressing the GES gene under the control of a heterologous promoter. GES transcription is upregulated under conditions that induce (E,E)-geranyllinalool and TMTT synthesis, including infestation of plants with larvae of the moth Plutella xylostella and treatment with the fungal peptide alamethicin or the octadecanoid mimic coronalon. Induction requires jasmonic acid but is independent from salicylic acid or ethylene. This study paves the ground to address the contribution of TMTT in ecological interactions and to elucidate the signaling network that regulates TMTT synthesis. PMID- 18398054 TI - KNOX lost the OX: the Arabidopsis KNATM gene defines a novel class of KNOX transcriptional regulators missing the homeodomain. AB - Three amino acid loop extension (TALE) homeodomain transcriptional regulators play a central role in plant and animal developmental programs. Plant KNOTTED1 like homeobox (KNOX) and animal Myeloid ecotropic viral integration site (MEIS) proteins share a TALE homeodomain and a MEINOX (MEIS-KNOX) domain, suggesting that an ancestral MEINOX-TALE protein predates the divergence of plants from fungi and animals. In this study, we identify and characterize the Arabidopsis thaliana KNATM gene, which encodes a MEINOX domain but not a homeodomain. Phylogenetic analysis of the KNOX family places KNATM in a new class and shows conservation in dicotyledons. We demonstrate that KNATM selectively interacts with Arabidopsis BELL TALE proteins through the MEINOX domain. The homeodomain is known to be necessary for KNOX-KNOX interaction. On the contrary, KNATM specifically dimerizes with the KNOX protein BREVIPEDICELLUS through an acidic coiled-coil domain. KNATM is expressed in proximal-lateral domains of organ primordia and at the boundary of mature organs; in accordance, genetic analyses identify a function for KNATM in leaf proximal-distal patterning. In vivo domain analyses highlighted KNATM functional regions and revealed a role as transcriptional regulator. Taken together, our data reveal a homeodomain independent mechanism of KNOX dimerization and transcriptional regulation. PMID- 18398055 TI - Stem-cell ecology and stem cells in motion. AB - This review highlights major scientific developments over the past 50 years or so in concepts related to stem-cell ecology and to stem cells in motion. Many thorough and eloquent reviews have been presented in the last 5 years updating progress in these issues. Some paradigms have been challenged, others validated, or new ones brought to light. In the present review, we will confine our remarks to the historical development of progress. In doing so, we will refrain from a detailed analysis of controversial data, emphasizing instead widely accepted views and some challenging novel ones. PMID- 18398056 TI - CD150- cells are transiently reconstituting multipotent progenitors with little or no stem cell activity. PMID- 18398058 TI - Dasatinib suppresses in vitro natural killer cell cytotoxicity. PMID- 18398059 TI - Single agent lenalidomide induces a response in refractory T-cell posttransplantation lymphoproliferative disorder. PMID- 18398060 TI - The American Society of Hematology: a success at age 50; blood banking and sodium citrate. PMID- 18398061 TI - MPL and JAK2 exon 12 mutations in patients with the Budd-Chiari syndrome or extrahepatic portal vein obstruction. PMID- 18398062 TI - Human cytomegalovirus DNAemia and preemptive treatment of CMV infection in children after hematopoietic stem-cell transplantation: is any question settled? PMID- 18398063 TI - Clarification of data used in three studies on MCA-induction of sarcoma in mice. PMID- 18398065 TI - Unrelated donor transplants in adults with Philadelphia-negative acute lymphoblastic leukemia in first complete remission. AB - We report the retrospective outcomes of unrelated donor (URD) transplants in 169 patients with acute lymphoblastic leukemia (ALL) in first complete remission (CR1) who received transplants between 1995 and 2004. Median age was 33 years (range, 16-59 years). A total of 50% had a white blood cell count (WBC) more than 30 x 10(9)/L, 18% extramedullary disease, 42% achieved CR more than 8 weeks from diagnosis, 25% had adverse cytogenetics, and 19% had T-cell leukemia. A total of 41% were HLA well-matched, 41% partially matched with their donors, and 18% were HLA-mismatched. At 54-month median follow-up, incidences of acute grade 2-IV, III to IV, and chronic graft-versus-host disease were 50%, 25%, and 43%, respectively. Five-year treatment-related mortality (TRM), relapse, and overall survival were 42%, 20%, and 39%, respectively. In multivariate analyses, TRM was significantly higher with HLA-mismatched donors and T-cell depletion. Relapse risk was higher if the diagnostic WBC was more than 100 x 10(9)/L. Factors associated with poorer survival included WBC more than 100 x 10(9)/L, more than 8 weeks to CR1, cytomegalovirus seropositivity, HLA mismatching, and T-cell depletion. Nearly 40% of adults with ALL in CR1 survive 5 years after URD transplantation. Relapse risks were modest; TRM is the major cause of treatment failure. Selecting closely HLA-matched URD and reducing TRM should improve results. PMID- 18398066 TI - RGT, a synthetic peptide corresponding to the integrin beta 3 cytoplasmic C terminal sequence, selectively inhibits outside-in signaling in human platelets by disrupting the interaction of integrin alpha IIb beta 3 with Src kinase. AB - Mutational analysis has established that the cytoplasmic tail of the integrin beta 3 subunit binds c-Src (termed as Src in this study) and is critical for bidirectional integrin signaling. Here we show in washed human platelets that a cell-permeable, myristoylated RGT peptide (myr-RGT) corresponding to the integrin beta 3 C-terminal sequence dose-dependently inhibited stable platelet adhesion and spreading on immobilized fibrinogen, and fibrin clot retraction as well. Myr RGT also inhibited the aggregation-dependent platelet secretion and secretion dependent second wave of platelet aggregation induced by adenosine diphosphate, ristocetin, or thrombin. Thus, myr-RGT inhibited integrin outside-in signaling. In contrast, myr-RGT had no inhibitory effect on adenosine diphosphate-induced soluble fibrinogen binding to platelets that is dependent on integrin inside-out signaling. Furthermore, the RGT peptide induced dissociation of Src from integrin beta 3 and dose-dependently inhibited the purified recombinant beta 3 cytoplasmic domain binding to Src-SH3. In addition, phosphorylation of the beta 3 cytoplasmic tyrosines, Y(747) and Y(759), was inhibited by myr-RGT. These data indicate an important role for beta 3-Src interaction in outside-in signaling. Thus, in intact human platelets, disruption of the association of Src with beta 3 and selective blockade of integrin alpha IIb beta 3 outside-in signaling by myr-RGT suggest a potential new antithrombotic strategy. PMID- 18398067 TI - The cover. Landscape. PMID- 18398068 TI - A piece of my mind. Wound healing. PMID- 18398069 TI - New tool measures 10-year fracture risk. PMID- 18398070 TI - Rapid stroke treatment an elusive goal. PMID- 18398071 TI - Scientists find clues to glucose intolerance. PMID- 18398072 TI - Excluding pulmonary embolism with computed tomographic pulmonary angiography or ventilation-perfusion lung scanning. PMID- 18398073 TI - Excluding pulmonary embolism with computed tomographic pulmonary angiography or ventilation-perfusion lung scanning. PMID- 18398074 TI - Evaluation of suspected pulmonary embolism during pregnancy. PMID- 18398075 TI - Prevalent vertebral fracture and the risk of incident vertebral fracture. PMID- 18398076 TI - Prevalent vertebral fracture and the risk of incident vertebral fracture. PMID- 18398077 TI - Regulating physician expert witness testimony. PMID- 18398078 TI - Regulating physician expert witness testimony. PMID- 18398079 TI - Chemoembolization combined with radiofrequency ablation for patients with hepatocellular carcinoma larger than 3 cm: a randomized controlled trial. AB - CONTEXT: Transarterial chemoembolization (TACE) combined with radiofrequency ablation (RFA) therapy has been used for patients with large hepatocellular carcinoma tumors, but the survival benefits of combined treatment are not known. OBJECTIVE: To compare rates of survival of patients with large hepatocellular carcinoma tumors who received treatment with TACE combined with RFA therapy (TACE RFA), TACE alone, and RFA alone. DESIGN, SETTING, AND PATIENTS: Randomized controlled trial conducted from January 2001 to May 2004 among 291 consecutive patients with hepatocellular carcinoma larger than 3 cm at a single center in China. INTERVENTION: Patients were randomly assigned to treatment with combined TACE-RFA (n = 96), TACE alone (n = 95), or RFA alone (n = 100). MAIN OUTCOME MEASURES: The primary end point was survival and the secondary end point was objective response rate. RESULTS: During a median 28.5 months of follow-up, median survival times were 24 months in the TACE group (3.4 courses), 22 months in the RFA group (3.6 courses), and 37 months in the TACE-RFA group (4.4 courses). Patients treated with TACE-RFA had better overall survival than those treated with TACE alone (hazard ratio [HR], 1.87; 95% confidence interval [CI], 1.33-2.63; P < .001) or RFA (HR, 1.88; 95% CI, 1.34-2.65; P < .001). In a preplanned substratification analysis, survival was also better in the TACE-RFA group than in the RFA group for patients with uninodular hepatocellular carcinoma (HR, 2.50; 95% CI, 1.42-4.42; P = .001) and in the TACE-RFA group than the TACE group for patients with multinodular hepatocellular carcinoma (HR, 1.99; 95% CI, 1.31-3.00; P < .001). The rate of objective response sustained for at least 6 months was higher in the TACE-RFA group (54%) than with either TACE (35%; rate difference, 0.19; 95% CI, 0.06-0.33; P = .009) or RFA (36%; rate difference, 0.18; 95% CI, 0.05-0.32; P = .01) treatment alone. CONCLUSION: In this patient group, TACE-RFA was superior to TACE alone or RFA alone in improving survival for patients with hepatocellular carcinoma larger than 3 cm. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00479050. PMID- 18398080 TI - Effect of lower targets for blood pressure and LDL cholesterol on atherosclerosis in diabetes: the SANDS randomized trial. AB - CONTEXT: Individuals with diabetes are at increased risk for cardiovascular disease (CVD), but more aggressive targets for risk factor control have not been tested. OBJECTIVE: To compare progression of subclinical atherosclerosis in adults with type 2 diabetes treated to reach aggressive targets of low-density lipoprotein cholesterol (LDL-C) of 70 mg/dL or lower and systolic blood pressure (SBP) of 115 mm Hg or lower vs standard targets of LDL-C of 100 mg/dL or lower and SBP of 130 mm Hg or lower. DESIGN, SETTING, AND PARTICIPANTS: A randomized, open-label, blinded-to-end point, 3-year trial from April 2003-July 2007 at 4 clinical centers in Oklahoma, Arizona, and South Dakota. Participants were 499 American Indian men and women aged 40 years or older with type 2 diabetes and no prior CVD events. INTERVENTIONS: Participants were randomized to aggressive (n=252) vs standard (n=247) treatment groups with stepped treatment algorithms defined for both. MAIN OUTCOME MEASURES: Primary end point was progression of atherosclerosis measured by common carotid artery intimal medial thickness (IMT). Secondary end points were other carotid and cardiac ultrasonographic measures and clinical events. RESULTS: Mean target LDL-C and SBP levels for both groups were reached and maintained. Mean (95% confidence interval) levels for LDL-C in the last 12 months were 72 (69-75) and 104 (101-106) mg/dL and SBP levels were 117 (115-118) and 129 (128-130) mm Hg in the aggressive vs standard groups, respectively. Compared with baseline, IMT regressed in the aggressive group and progressed in the standard group (-0.012 mm vs 0.038 mm; P < .001); carotid arterial cross-sectional area also regressed (-0.02 mm(2) vs 1.05 mm(2); P < .001); and there was greater decrease in left ventricular mass index (-2.4 g/m(2.7) vs -1.2 g/m(2.7); P = .03) in the aggressive group. Rates of adverse events (38.5% and 26.7%; P = .005) and serious adverse events (n = 4 vs 1; P = .18) related to blood pressure medications were higher in the aggressive group. Clinical CVD events (1.6/100 and 1.5/100 person-years; P = .87) did not differ significantly between groups. CONCLUSIONS: Reducing LDL-C and SBP to lower targets resulted in regression of carotid IMT and greater decrease in left ventricular mass in individuals with type 2 diabetes. Clinical events were lower than expected and did not differ significantly between groups. Further follow-up is needed to determine whether these improvements will result in lower long-term CVD event rates and costs and favorable risk-benefit outcomes. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00047424. PMID- 18398081 TI - Omega-3 free fatty acids for the maintenance of remission in Crohn disease: the EPIC Randomized Controlled Trials. AB - CONTEXT: Maintenance therapy for Crohn disease features the use of immunosuppressive drugs, which are associated with an increased risk of infection. Identification of safe and effective maintenance strategies is a priority. OBJECTIVE: To determine whether the oral administration of omega-3 free fatty acids is more effective than placebo for prevention of relapse of Crohn disease. DESIGN, SETTING, AND PATIENTS: Two randomized, double-blind, placebo controlled studies (Epanova Program in Crohn's Study 1 [EPIC-1] and EPIC-2) conducted between January 2003 and February 2007 at 98 centers in Canada, Europe, Israel, and the United States. Data from 363 and 375 patients with quiescent Crohn disease were evaluated in EPIC-1 and EPIC-2, respectively. INTERVENTIONS: Patients with a Crohn's Disease Activity Index (CDAI) score of less than 150 were randomly assigned to receive either 4 g/d of omega-3 free fatty acids or placebo for up to 58 weeks. No other treatments for Crohn disease were permitted. MAIN OUTCOME MEASURE: Clinical relapse, as defined by a CDAI score of 150 points or greater and an increase of more than 70 points from the baseline value, or initiation of treatment for active Crohn disease. RESULTS: For EPIC-1, 188 patients were assigned to receive omega-3 free fatty acids and 186 patients to receive placebo. Corresponding numbers for EPIC-2 were 189 and 190 patients, respectively. The rate of relapse at 1 year in EPIC-1 was 31.6% in patients who received omega-3 free fatty acids and 35.7% in those who received placebo (hazard ratio, 0.82; 95% confidence interval, 0.51-1.19; P = .30). Corresponding values for EPIC-2 were 47.8% and 48.8% (hazard ratio, 0.90; 95% confidence interval, 0.67-1.21; P = .48). Serious adverse events were uncommon and mostly related to Crohn disease. CONCLUSION: In these trials, treatment with omega-3 free fatty acids was not effective for the prevention of relapse in Crohn disease. TRIAL REGISTRATION: clinicaltrials.gov Identifiers: EPIC-1: NCT00613197, EPIC-2: NCT00074542. PMID- 18398082 TI - Effectiveness of specialized palliative care: a systematic review. AB - CONTEXT: Specialized palliative care teams are increasingly providing care for the terminally ill. However, the impact of such teams on quality of life, satisfaction with care, and economic cost has not been examined systematically using detailed criteria for study quality. OBJECTIVE: To systematically review the evidence for effectiveness of specialized palliative care. DATA SOURCES: We performed a keyword search of the following databases from their inception to January 2008: MEDLINE, Ovid Healthstar, CINAHL, EMBASE, and the Cochrane Central Register of Controlled Trials. STUDY SELECTION: We included all randomized controlled trials in which specialized palliative care was the intervention and for which outcomes included quality of life, satisfaction with care, or economic cost. DATA EXTRACTION: Data on population, intervention, outcome, methods, and methodological quality were extracted by 2 investigators using standardized criteria. RESULTS: Of 396 reports of randomized controlled trials, 22 met our inclusion criteria. There was most consistent evidence for effectiveness of specialized palliative care in improvement of family satisfaction with care (7 of 10 studies favored the intervention). Only 4 of 13 studies assessing quality of life and 1 of 14 assessing symptoms showed a significant benefit of the intervention; however, most studies lacked statistical power to report conclusive results, and quality-of-life measures were not specific for terminally ill patients. There was evidence of significant cost savings of specialized palliative care in only 1 of the 7 studies that assessed this outcome. Methodological limitations were identified in all trials, including contamination of the control group, failure to account for clustering in cluster randomization studies, and substantial problems with recruitment, attrition, and adherence. CONCLUSIONS: The evidence for benefit from specialized palliative care is sparse and limited by methodological shortcomings. Carefully planned trials, using a standardized palliative care intervention and measures constructed specifically for this population, are needed. PMID- 18398083 TI - When should data and safety monitoring committees share interim results in cardiovascular trials? PMID- 18398084 TI - The Iraqi refugee crisis: familiar problems and new challenges. PMID- 18398085 TI - Expanding the treatment options for hepatocellular carcinoma: combining transarterial chemoembolization with radiofrequency ablation. PMID- 18398086 TI - The great debate of 2008--how low to go in preventive cardiology? PMID- 18398087 TI - JAMA patient page. Crohn disease. PMID- 18398088 TI - Yuan Chang and Patrick Moore: teaming up to hunt down cancer-causing viruses. PMID- 18398089 TI - Filling the gaps to fight cervical cancer. PMID- 18398090 TI - Hepatitis B virus-induced liver cancer in Asian Americans: a preventable disease. PMID- 18398091 TI - Doctors seek to prevent breast cancer recurrence by lowering insulin levels. PMID- 18398092 TI - Stat bite: Estimated new leukemia cases in 2008. PMID- 18398093 TI - Can early response assessment guide neoadjuvant chemotherapy in early-stage breast cancer? PMID- 18398094 TI - Intensified neoadjuvant chemotherapy in early-responding breast cancer: phase III randomized GeparTrio study. AB - BACKGROUND: Patients with an early response to neoadjuvant chemotherapy have chemosensitive tumors and a high probability for a pathological complete response at surgery. The relationship between extended chemotherapy and pathological complete response at surgery was investigated in a clinical trial. METHODS: Untreated breast cancer patients received two 3-week cycles of docetaxel at 75 mg/m(2), doxorubicin at 50 mg/m(2), and cyclophosphamide at 500 mg/m(2) (TAC). Those whose tumor size decreased by 50% or more by sonographic measurement (ie, reduction in the product of the two largest perpendicular diameters by at least 50%) were classified as responders and randomly assigned to receive four or six more cycles of TAC, for a total of six or eight TAC cycles. The primary aim was to increase the rate of a pathological complete response (defined as no invasive or in situ residual tumor masses in the breast and lymph nodes) from 20% to 26%. Sonographic response rates and rates of breast-conserving surgery and adverse effects were also assessed. All statistical tests were two-sided. RESULTS: Of the 2090 patients in the GeparTrio trial, 1390 (66.5%) were randomly assigned as responders after two initial TAC cycles to receive an additional four (n = 704) or six (n = 686) TAC cycles. Rates of pathological complete response were not statistically significantly different between the arms (21.0% with six TAC cycles and 23.5% with eight TAC cycles; difference = 2.5%, 95% confidence interval [CI] = -1.8% to 6.8%; P = .27). More clinical (48.2% vs 52.9%, difference = 4.7%; 95% CI = -0.55% to 9.95%; P = .08) and sonographic (22.6% vs 27.6%, difference = 5%; 95% CI = 0.45% to 9.55%; P = .033) complete responses at surgery were observed with eight TAC cycles than with six TAC cycles. The rate of breast-conserving surgery was similar in both arms (67.5% vs 68.5%, respectively, P = .68). Grade 3 or 4 leukopenia and edema and various grade 1 or 2 adverse events were more frequent in patients receiving eight TAC cycles than in those receiving six cycles. CONCLUSION: Patients receiving eight TAC cycles had statistically significantly higher sonographic response rates but not pathological complete response rates than those receiving six TAC cycles. However, they also had more toxic effects. So far, eight cycles of TAC cannot be recommended for the whole group of patients responding to two initial cycles of TAC. PMID- 18398096 TI - Re: Second cancers among 104,760 survivors of cervical cancer: evaluation of long term risk. PMID- 18398095 TI - A threefold dose intensity treatment with ifosfamide, carboplatin, and etoposide for patients with small cell lung cancer: a randomized trial. AB - BACKGROUND: The dose intensity of chemotherapy can be increased to the highest possible level by early administration of multiple and sequential high-dose cycles supported by transfusion with peripheral blood progenitor cells (PBPCs). A randomized trial was performed to test the impact of such dose intensification on the long-term survival of patients with small cell lung cancer (SCLC). METHODS: Patients who had limited or extensive SCLC with no more than two metastatic sites were randomly assigned to high-dose (High, n = 69) or standard-dose (Std, n = 71) chemotherapy with ifosfamide, carboplatin, and etoposide (ICE). High-ICE cycles were supported by transfusion with PBPCs that were collected after two cycles of treatment with epidoxorubicin at 150 mg/m(2), paclitaxel at 175 mg/m(2), and filgrastim. The primary outcome was 3-year survival. Comparisons between response rates and toxic effects within subgroups (limited or extensive disease, liver metastases or no liver metastases, Eastern Cooperative Oncology Group performance status of 0 or 1, normal or abnormal lactate dehydrogenase levels) were also performed. RESULTS: Median relative dose intensity in the High-ICE arm was 293% (range = 174%-392%) of that in the Std-ICE arm. The 3-year survival rates were 18% (95% confidence interval [CI] = 10% to 29%) and 19% (95% CI = 11% to 30%) in the High-ICE and Std-ICE arms, respectively. No differences were observed between the High-ICE and Std-ICE arms in overall response (n = 54 [78%, 95% CI = 67% to 87%] and n = 48 [68%, 95% CI = 55% to 78%], respectively) or complete response (n = 27 [39%, 95% CI = 28% to 52%] and n = 24 [34%, 95% CI = 23% to 46%], respectively). Subgroup analyses showed no benefit for any outcome from High-ICE treatment. Hematologic toxicity was substantial in the Std-ICE arm (grade > or = 3 neutropenia, n = 49 [70%]; anemia, n = 17 [25%]; thrombopenia, n = 17 [25%]), and three patients (4%) died from toxicity. High-ICE treatment was predictably associated with severe myelosuppression, and five patients (8%) died from toxicity. CONCLUSIONS: The long-term outcome of SCLC was not improved by raising the dose intensity of ICE chemotherapy by threefold. PMID- 18398097 TI - Neoadjuvant vinorelbine-capecitabine versus docetaxel-doxorubicin cyclophosphamide in early nonresponsive breast cancer: phase III randomized GeparTrio trial. AB - BACKGROUND: Among breast cancer patients, nonresponse to initial neoadjuvant chemotherapy is associated with unfavorable outcome. We compared the response of nonresponding patients who continued the same treatment with that of patients who switched to a well-tolerated non-cross-resistant regimen. METHODS: Previously untreated breast cancer patients received two 3-week cycles of docetaxel at 75 mg/m(2), doxorubicin at 50 mg/m(2), and cyclophosphamide at 500 mg/m(2) per day (TAC). Patients whose tumors did not decrease in size by at least 50% were randomly assigned to four additional cycles of TAC or to four cycles of vinorelbine at 25 mg/m(2) and capecitabine at 2000 mg/m(2) (NX). The outcome was sonographic response, defined as a reduction in the product of the two largest perpendicular diameters by at least 50%. A difference of 10% or less in the sonographic response qualified as noninferiority of the NX treatment. Pathological complete response was defined as no invasive or in situ residual tumor masses in the breast and lymph nodes. Toxic effects were assessed. All statistical tests were two-sided. RESULTS: Of 2090 patients enrolled in the GeparTrio study, 622 (29.8%) who did not respond to two initial cycles of TAC were randomly assigned to an additional four cycles of TAC (n = 321) or to four cycles of NX (n = 301). Sonographic response rate was 50.5% for the TAC arm and 51.2% for the NX arm. The difference of 0.7% (95% confidence interval = -7.1% to 8.5%) demonstrated noninferiority of NX (P = .008). Similar numbers of patients in both arms received breast-conserving surgery (184 [57.3%] in the TAC arm vs 180 [59.8%] in the NX arm) and had a pathological complete response (5.3% vs 6.0%). Fewer patients in the NX arm than in the TAC arm had hematologic toxic effects, mucositis, infections, and nail changes, but more had hand-foot syndrome and sensory neuropathy. CONCLUSION: Pathological complete responses to both regimens were marginal. Among patients who did not respond to the initial neoadjuvant TAC treatment, similar efficacy but better tolerability was observed by switching to NX than continuing with TAC. PMID- 18398098 TI - Donation frequency, iron loss, and risk of cancer among blood donors. AB - BACKGROUND: Long-term deleterious effects of repeated blood donations may be masked by the donors' healthy lifestyle. To investigate possible effects of blood donation and iron loss through blood donation on cancer incidence while minimizing "healthy donor effects," we made dose-response comparisons within a cohort of Swedish and Danish blood donors. METHODS: We used a nested case-control study design, in which case patients were defined as all donors who were diagnosed with a malignancy between their first recorded blood donation and study termination (n = 10866). Control subjects (n = 107140) were individually matched on sex, age, and county of residence. Using conditional logistic regression, we estimated relative risks of cancer according to number of blood donations made or estimated iron loss 3-12 years before a case patient was diagnosed with cancer. All statistical tests were two-sided. RESULTS: No clear association was observed between number of donations and risk of cancer overall. However, between the lowest (< or = median, < 0.75 g) and highest (> 90th percentile, > 2.7 g) categories of estimated iron loss, there was a trend (P(trend) < .001) of decreasing risk for cancers of the liver, lung, colon, stomach, and esophagus, which are thought to be promoted by iron overload (combined odds ratio [OR] = 0.70, 95% confidence interval [CI] = 0.58 to 0.84), but only among men and only with a latency of 3-7 years. The risk of non-Hodgkin lymphoma was higher among frequent plasma donors (> 25 vs 0 donations, OR = 2.14, 95% CI = 1.22 to 3.74). CONCLUSIONS: Repeated blood donation was not associated with increased or decreased risk of cancer overall. The lack of consistency across latency periods casts doubt on an apparent association between reduced cancer risk and iron loss in men. The positive association between frequent plasma donation and risk of non Hodgkin lymphoma deserves further exploration. PMID- 18398099 TI - Diseases desperate grown. PMID- 18398100 TI - Re: Declines in invasive breast cancer and use of postmenopausal hormone therapy in a screening mammography population. PMID- 18398101 TI - Re: Declines in invasive breast cancer and use of postmenopausal hormone therapy in a screening mammography population. PMID- 18398102 TI - Re: Breast cancer incidence, 1980-2006: combined roles of menopausal hormone therapy, screening mammography, and estrogen receptor status. PMID- 18398103 TI - Re: Breast cancer incidence, 1980-2006: combined roles of menopausal hormone therapy, screening mammography, and estrogen receptor status. PMID- 18398104 TI - Mechanism of synergy of N-(4-hydroxyphenyl)retinamide and ABT-737 in acute lymphoblastic leukemia cell lines: Mcl-1 inactivation. AB - BACKGROUND: ABT-737 is a pan-Bcl-2 inhibitor that has a wide range of single agent activity against acute lymphoblastic leukemia (ALL) cell lines and xenografts. A relationship between expression of myeloid cell leukemia 1 (Mcl-1), an antiapoptotic member of the Bcl-2 family of proteins, and resistance to ABT 737 has been reported for various cancers. The synthetic cytotoxic retinoid N-(4 hydroxyphenyl)retinamide (4-HPR) is known to generate reactive oxygen species (ROS), and ROS have been shown to activate c-Jun kinase (JNK), which in turn phosphorylates and inhibits Mcl-1. Thus, we investigated whether 4-HPR-mediated inactivation of Mcl-1 could act synergistically with ABT-737 to promote leukemia cell death. METHODS: Cytotoxicity was determined using the fluorescence-based DIMSCAN assay. Synergy was defined as a combination index (CIN) less than 1. The expression of Bcl-2 family messenger RNAs was measured by real-time reverse transcription-polymerase chain reaction, and caspase activity was measured enzymatically. Changes in Bcl-2 family proteins and release of mitochondrial cytochrome c were detected by immunoblotting. ROS, apoptosis, mitochondrial membrane depolarization, and phospho-JNK were measured by flow cytometry. Gene silencing was by small interfering RNA (siRNA). All statistical tests were two sided. RESULTS: ABT-737 decreased Mcl-1 protein expression in ABT-737-sensitive ALL cell lines but not in ABT-737-resistant lines. Using the antioxidant ascorbic acid and siRNA-mediated knockdown of JNK, we showed that 4-HPR decreased Mcl-1 via ROS generation (that phosphorylates JNK) in ABT-737-resistant cell lines. Combining ABT-737 with 4-HPR enhanced the mitochondrial apoptotic cascade (percentage of cells with depolarized mitochondrial membrane at 6 hours, ABT-737 vs ABT-737 plus 4-HPR: 24.5% vs 45.5%, difference = 20.1%, 95% CI = 18.9% to 13.9%; P < .001) and caused caspase-dependent, synergistic multilog cytotoxicity in all seven ALL cell lines examined (mean CIN = 0.57, 95% CI = 0.37 to 0.87), with minimal cytotoxicity for normal lymphocytes. CONCLUSIONS: An increase of Mcl 1 protein in response to ABT-737 is one mechanism of ABT-737 resistance that can be overcome by 4-HPR, resulting in synergistic cytotoxicity of ABT-737 combined with 4-HPR in ALL cell lines. PMID- 18398105 TI - Conjugated equine estrogen and risk of benign proliferative breast disease: a randomized controlled trial. AB - BACKGROUND: Estrogens stimulate proliferation of breast epithelium and may therefore increase the risk of benign proliferative breast disease, a condition that is associated with increased risk of breast cancer. We tested the effect of conjugated equine estrogen (CEE) on risk of benign proliferative breast disease in the Women's Health Initiative (WHI) randomized controlled trial. METHODS: In the WHI CEE trial, 10,739 postmenopausal women were randomly assigned to 0.625 mg/d of CEE or to placebo. Baseline and annual breast examinations and mammograms were required. We identified women in the trial who reported breast biopsies that were free of cancer, obtained the associated histological sections, and subjected them to standardized central review. Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: A total of 232 incident cases of benign proliferative breast disease were ascertained during follow-up (average duration, 6.9 years), with 155 in the CEE group and 77 in the placebo group. Use of CEE was associated with a more than two-fold increase in the risk of benign proliferative breast disease (HR = 2.11, 95% CI = 1.58 to 2.81). For benign proliferative breast disease without atypia, the HR was 2.34 (95% CI = 1.71 to 3.20), whereas for atypical hyperplasia, it was 1.12 (95% CI = 0.53 to 2.40). Risk varied little by levels of baseline characteristics. CONCLUSION: Use of 0.625 mg/d of CEE was associated with a statistically significant increased risk of benign proliferative breast disease. PMID- 18398107 TI - Calpain in the CNS: from synaptic function to neurotoxicity. AB - The calpains are a class of cellular cysteine proteases that require calcium and are functionally active at neutral pH. Calpain activation can take place in two modes: controlled activation under physiological conditions (in which only a few molecules of calpain are activated per cell), and hyperactivation under pathological conditions that involve sustained calcium overload (in which all available calpain molecules are activated). Regulated activation of calpain in the central nervous system (CNS) may be critical to synaptic function and memory formation, with possible substrates including various structural and scaffolding proteins, enzymes, and glutamate receptors. Hyperactivation of calpain in the central nervous system is generally associated with severe cellular challenge or damage. Calpain cleavage products may thus provide useful biomarkers for the presence of neurodegenerative processes or neuronal injury. PMID- 18398108 TI - Assessment of aortic pulse wave velocity and cardiac diastolic function in subjects with and without the metabolic syndrome: HDL cholesterol is independently associated with cardiovascular function. AB - OBJECTIVE: To evaluate the influence of lipid and glucose metabolism in the metabolic syndrome on aortic pulse wave velocity (PWV) and left ventricular (LV) diastolic function using magnetic resonance imaging (MRI). RESEARCH DESIGN AND METHODS: Aortic PWV and LV diastolic function were assessed using MRI in 16 subjects with the metabolic syndrome and 16 subjects without the metabolic syndrome matched for age, waist circumference, and blood pressure. The groups were compared using the unpaired t test or Mann-Whitney U test, and linear regression analysis was applied. RESULTS: Aortic PWV was increased and LV diastolic function was decreased in subjects with compared with those without the metabolic syndrome. HDL cholesterol was independently associated with aortic PWV (R = -0.470, P < 0.01) and LV diastolic function (R = -0.421, P = 0.02). CONCLUSIONS: Increased aortic PWV and decreased LV diastolic function is observed in subjects with the metabolic syndrome, regardless of blood pressure. Moreover, HDL cholesterol is independently associated with aortic PWV and LV diastolic function. PMID- 18398109 TI - Aerosolized beta-adrenergic agonist therapy reduces pulmonary edema following lung surgery. PMID- 18398110 TI - An ounce of tuberculosis prevention. PMID- 18398111 TI - Ultrasound: the pulmonologist's new best friend. PMID- 18398112 TI - Exhaled nitric oxide in asthma. From diagnosis, to monitoring, to screening: are we there yet? PMID- 18398113 TI - The hypoxia altitude simulation test: an increasingly performed test for the evaluation of patients prior to air travel. PMID- 18398114 TI - Affording endobronchial ultrasound. PMID- 18398116 TI - Anti-endothelial cell antibodies in patients with sarcoidosis. AB - BACKGROUND: Anti-endothelial cell antibodies (AECA) are circulating antibodies that bind to endothelial antigens and induce endothelial cell damage. These antibodies have been detected in patients with collagen vascular disease and systemic vasculitis. Sarcoidosis is a multiple granulomatous disorder of unknown etiology, and its clinical presentation, organ involvement, and prognosis are highly diverse. In this study, we aimed to investigate the expression of AECA in patients with sarcoidosis. We also examined whether these antibodies could serve as a marker for the activity, severity, and prognosis of sarcoidosis. METHODS: Forty sarcoidosis patients, whose diagnosis was established by clinicoradiologic findings and histologic confirmation of noncaseating epithelioid cell granulomas, were studied. Serum and BAL samples were examined for AECA by cellular enzyme linked immunosorbent assay (ELISA) using human umbilical vein endothelial cells. The findings were expressed in terms of an ELISA ratio (ER). Fifty-seven subjects without any clinical, radiologic, or serologic evidence of pulmonary disease or autoimmune disorders served as the reference population to judge the positivity of AECA. RESULTS: Patients with sarcoidosis had a significantly higher positivity rate for and levels of AECA in both serum and BAL fluid than the reference population. In addition, the ER of AECA was significantly elevated in patients with multiple lesions or who required corticosteroid therapy compared with that in patients without multiple lesions or who did not need corticosteroid therapy. CONCLUSION: Expression of AECA might be a useful marker to predict the course of sarcoidosis. PMID- 18398117 TI - Surfactant in airway disease. AB - Beta(2)-adrenergic agonists cause a release of pulmonary surfactant into lung airways. The surfactant phospholipids maintain the patency of the conducting airways, but this function is inhibited by plasma proteins entering an inflamed airway. The physical behavior of the surfactant can be studied with a pulsating bubble surfactometer and a capillary surfactometer. Calf lung surfactant extract was found to be inhibited by plasma proteins and by a lowering of temperature. Severe breathing difficulties and malfunctioning surfactant developed in BALB/c mice inhaling ozone or infected with respiratory syncytial virus, mainly as a result of proteins invading the airways. Patients with asthma were challenged with allergens in an area of one lung. BAL fluid (BALF) from such an area contained a surfactant that functioned poorly (ie, an inability to maintain airway openness) compared with BALF from the other lung or from the lungs of healthy volunteers. When proteins in the BALF were removed, surfactant performance clearly improved. Eosinophils, so prominent in asthmatic patients, synthesize the enzyme lysophospholipase, which, together with the enzyme phospholipase A(2), catalyzes the hydrolysis of the main component of the surfactant, phosphatidylcholine. Such hydrolysis incapacitates the ability of the surfactant to maintain airway patency. The treatment of asthma with beta(2) adrenergic agonists and steroids will have a valuable effect on the surfactant system. It will cause a release of fresh surfactant into terminal airways. Surfactant can also be nebulized and inhaled, which has been shown to be an effective treatment. PMID- 18398118 TI - Screening and familial testing of patients for alpha 1-antitrypsin deficiency. AB - alpha(1)-Antitrypsin deficiency (AATD) is an autosomal-codominant genetic disorder that predisposes individuals to the development of liver and lung disease. AATD is greatly underrecognized and underdiagnosed. Early identification allows preventive measures to be taken, the most important of which is the avoidance of smoking (including the inhalation of second-hand smoke) and exposure to environmental pollutants. Early detection also allows careful lung function monitoring and augmentation therapy while the patient still has preserved lung function. Cost factors and controversies have discouraged the initiation of large scale screening programs of the newborn and adult populations in the United States and Europe (except for Sweden). There are sound medical reasons for targeted screening. Evidence-based recommendations for testing have been published by the American Thoracic Society/European Respiratory Society task force, which take potential social, psychological, and ethical adverse factors into consideration. This review discusses rationales for testing and screening for AATD in asymptomatic individuals, family members, and the general population, weighing benefits against potential psychological, social, and ethical implications of testing. For most, negative issues are outweighed by the benefits of testing. AATD testing should be routine in the management of adults with emphysema, COPD, and asthma with incompletely reversible airflow obstruction. PMID- 18398119 TI - Targeting airway inflammation in asthma: current and future therapies. AB - Asthma is a chronic inflammatory disease of the airway that requires long-term antiinflammatory therapy. Inhaled corticosteroids (ICSs) are recommended for first-line treatment of persistent disease, but not all patients achieve asthma control even when these agents are used in high doses and in combination with other medications, including a long-acting beta(2)-agonist or a leukotriene modifier. Such patients may require additional therapy. As information about asthma pathophysiology and inflammatory phenotypes continues to increase, and additional antiinflammatory options become available, it may be possible to target antiinflammatory therapy to various aspects of the disease and consequently to improve the treatment of patients with inadequate responses to standard ICS-based therapy. Several novel antiinflammatory therapies are in different stages of clinical development. The most clinically advanced of these is omalizumab, a recombinant humanized monoclonal antibody that specifically targets IgE and is indicated for patients with moderate-to-severe asthma caused by allergies. Omalizumab has demonstrated efficacy in patients with moderate-to severe asthma and documented evidence of allergen sensitivity. Other key therapy options in clinical development either target proinflammatory cytokines (eg, interleukin-4 and tumor necrosis factor-alpha) or inflammatory cells (eg, T helper type 2 cells and eosinophils). This review provides an overview of the current and future approaches targeting airway inflammation in patients with asthma. PMID- 18398120 TI - Unilateral lymphocytic pleuritis as a manifestation of familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disease affecting predominantly populations surrounding the Mediterranean basin. It is the most prevalent hereditary periodic fever syndrome characterized mainly by recurrent and short attacks of fever and serositis (pleuritis, arthritis, peritonitis). Unilateral polymorphonuclear exudative pleuritis associated with fever has been reported as the solitary manifestation of the first FMF attack, in < 10% of patients. This case study describes a 30-year-old Greek man with recurrent episodes of lymphocytic exudative pleuritis associated with fever. After a thorough workup (clinical criteria and molecular genetic testing identifying homozygosity polymorphisms of the FMF gene), the diagnosis of FMF was established. Treatment with colchicine, 2 mg/d, eliminated FMF attacks. To our knowledge, this is the first well-documented case report of a patient with FMF presenting with a lymphocytic exudative pleural effusion. PMID- 18398121 TI - Hypoxia altitude simulation test. AB - A large number of patients with underlying pulmonary disease travel by air each year and are therefore at risk for significant cardiopulmonary effects of induced hypoxia at higher altitudes. The hypoxia altitude simulation test provides a simple way to identify those patients at risk by simulating conditions encountered at high altitude. By asking the patient to breathe a mixture of gases with an oxygen saturation of 15.1%, the test simulates a cabin pressure of 8,000 feet and allows the physician to screen for hypoxia, significant symptoms, and arrhythmias. Repeating the test with supplemental oxygen ensure adequate treatment of those patients who have a decrease in the alveolar pressure of oxygen, significant symptoms, and/or arrhythmias. PMID- 18398122 TI - Toxicology in the critical care unit. AB - Toxicologic conditions are encountered in critically ill patients due to intentional or unintentional misuse of or exposure to therapeutic or illicit drugs. Additionally, toxicities related to medical interventions may develop in hospitalized patients. This review focuses on recent developments in the field of critical care toxicology. Early interventions to decrease absorption or enhance elimination of toxins have limited value. Specific interventions to manage toxicities due to analgesics, sedative-hypnotics, antidepressants, antipsychotics, cardiovascular agents, alcohols, carbon monoxide, and cholinergic agents are reviewed. Hospital-acquired toxicities due to methemoglobinemia, propylene glycol, and propofol should be recognized and treated. The clinician is continually required to incorporate clinical judgment along with available scientific data and clinical evidence to determine the best therapy for toxicologic conditions. PMID- 18398123 TI - A 75-year-old woman with dyspnea and a sore throat. PMID- 18398124 TI - A 57-year-old man with end-stage renal disease and chronic cough. PMID- 18398125 TI - A 35-year-old man with recurrent pneumonias, eczema, coarse facial features, and cystic lung lesions. PMID- 18398126 TI - A 29-year-old immunocompetent man with meningitis and a large pulmonary mass. PMID- 18398127 TI - Chronic dyspnea and wheezing in a 46-year-old nonsmoker. PMID- 18398128 TI - Creating written patient education materials. PMID- 18398129 TI - A 34-year-old man presenting with gradual onset of shortness of breath and interstitial lung disease. PMID- 18398132 TI - There are major problems with the American College of Chest Physicians Second Lung Cancer Guidelines. PMID- 18398133 TI - Informed assent and medical education. PMID- 18398134 TI - Apical and midventricular transient left ventricular dysfunction syndrome (tako tsubo cardiomyopathy). PMID- 18398135 TI - Utility of CT for predicting pulmonary hypertension in patients with parenchymal lung disease: similar results, different conclusion? PMID- 18398136 TI - Hypopnea scoring in the Sleep Heart Health Study. PMID- 18398137 TI - Remembering other causes of alveolar siderophages: macrophage activation syndrome. PMID- 18398138 TI - Weak proinsulin peptide-major histocompatibility complexes are targeted in autoimmune diabetes in mice. AB - OBJECTIVE: Weak major histocompatibility complex (MHC) binding of self-peptides has been proposed as a mechanism that may contribute to autoimmunity by allowing for escape of autoreactive T-cells from the thymus. We examined the relationship between the MHC-binding characteristics of a beta-cell antigen epitope and T-cell autoreactivity in a model of autoimmune diabetes. RESEARCH DESIGN AND METHODS: The binding of a proinsulin epitope, proinsulin-1(47-64) (PI-1[47-64]), to the MHC class II molecules I-A(g7) and I-A(k) was measured using purified class II molecules. T-cell reactivity to the proinsulin epitope was examined in I-A(g7+) and I-A(k+) mice. RESULTS: C-peptide epitopes bound very weakly to I-A(g7) molecules. However, C-peptide-reactive T-cells were induced after immunization in I-A(g7)-bearing mice (NOD and B6.g7) but not in I-A(k)-bearing mice (B10.BR and NOD.h4). T-cells reactive with the PI-1(47-64) peptide were found spontaneously in the peripancreatic lymph nodes of pre-diabetic NOD mice. These T-cells were activated by freshly isolated beta-cells in the presence of antigen-presenting cells and caused diabetes when transferred into NOD.scid mice. CONCLUSIONS: These data demonstrate an inverse relationship between self-peptide-MHC binding and T cell autoreactivity for the PI-1(47-64) epitope in autoimmune diabetes. PMID- 18398139 TI - Glucose metabolism in vivo in four commonly used inbred mouse strains. AB - OBJECTIVE: To characterize differences in whole-body glucose metabolism between commonly used inbred mouse strains. RESEARCH DESIGN AND METHODS: Hyperinsulinemic euglycemic (approximately 8.5 mmol/l) and -hypoglycemic (approximately 3.0 mmol/l) clamps were done in catheterized, 5-h-fasted mice to assess insulin action and hypoglycemic counter-regulatory responsiveness. Hyperglycemic clamps (approximately 15 mmol/l) were done to assess insulin secretion and compared with results in perifused islets. RESULTS: Insulin action and hypoglycemic counter regulatory and insulin secretory phenotypes varied considerably in four inbred mouse strains. In vivo insulin secretion was greatest in 129X1/Sv mice, but the counter-regulatory response to hypoglycemia was blunted. FVB/N mice in vivo showed no increase in glucose-stimulated insulin secretion, relative hepatic insulin resistance, and the highest counter-regulatory response to hypoglycemia. In DBA/2 mice, insulin action was lowest among the strains, and islets isolated had the greatest glucose-stimulated insulin secretion in vitro. In C57BL/6 mice, in vivo physiological responses to hyperinsulinemia at euglycemia and hypoglycemia were intermediate relative to other strains. Insulin secretion by C57BL/6 mice was similar to that in other strains in contrast to the blunted glucose-stimulated insulin secretion from isolated islets. CONCLUSIONS: Strain dependent differences exist in four inbred mouse strains frequently used for genetic manipulation and study of glucose metabolism. These results are important for selecting inbred mice to study glucose metabolism and for interpreting and designing experiments. PMID- 18398140 TI - Hormone-sensitive lipase serine phosphorylation and glycerol exchange across skeletal muscle in lean and obese subjects: effect of beta-adrenergic stimulation. AB - OBJECTIVE: Increased intramuscular triacylglycerol (IMTG) storage is a characteristic of the obese insulin-resistant state. We aimed to investigate whether a blunted fasting or beta-adrenergically mediated lipolysis contributes to this increased IMTG storage in obesity. RESEARCH DESIGN AND METHODS: Forearm skeletal muscle lipolysis was investigated in 13 lean and 10 obese men using [(2)H(5)]glycerol combined with the measurement of arteriovenous differences before and during beta-adrenergic stimulation using the nonselective beta-agonist isoprenaline (ISO). Muscle biopsies were taken from the vastus lateralis muscle before and during ISO to investigate hormone-sensitive lipase (HSL) protein expression and serine phosphorylation. RESULTS: Baseline total glycerol release across the forearm was significantly blunted in obese compared with lean subjects (P = 0.045). This was accompanied by lower HSL protein expression (P = 0.004), HSL phosphorylation on PKA sites Ser(563) (P = 0.041) and Ser(659) (P = 0.09), and HSL phosphorylation on the AMPK site Ser(565) (P = 0.007), suggesting a blunted skeletal muscle lipolysis in obesity. Total forearm glycerol uptake during baseline did not differ significantly between groups, whereas higher net fatty acid uptake across the forearm was observed in the obese (P = 0.064). ISO induced an increase in total glycerol release from skeletal muscle, which was not significantly different between groups. Interestingly, this was accompanied by an increase in HSL Ser(659) phosphorylation in obese subjects during ISO compared with baseline (P = 0.008). CONCLUSIONS: Obesity is accompanied by impaired fasting glycerol release, lower HSL protein expression, and serine phosphorylation. It remains to be determined whether this is a primary factor or an adaptation to the obese insulin-resistant state. PMID- 18398141 TI - Monoclonal antibodies in cancer therapy: 25 years of progress. PMID- 18398142 TI - Defining the optimal schedule of drug administration: art or science? PMID- 18398143 TI - Bleomycin in testicular cancer: will pharmacogenomics improve treatment regimens? PMID- 18398144 TI - Perioperative risks of bevacizumab and other biologic agents for hepatectomy: theoretical or evidence based? PMID- 18398145 TI - Phase I study of a novel capecitabine schedule based on the Norton-Simon mathematical model in patients with metastatic breast cancer. AB - PURPOSE: This study was conducted to determine, in patients with advanced-stage breast cancer, the maximum tolerated dose (MTD) of capecitabine administered orally for 7 days followed by a 7-day rest (7/7), a schedule based on a mathematical method for the optimization of anticancer drug scheduling. PATIENTS AND METHODS: Eligible patients had measurable, metastatic breast cancer. There was no limit to number of prior treatments. A standard, three-patients-per-cohort dose-escalation scheme used flat-dose capecitabine beginning at 1,500 mg orally twice daily (bid) on a 7/7 schedule. Each cohort was monitored for 28 days before escalation to the next cohort to assess for delayed toxicity. Response was evaluated radiographically every 12 weeks; toxicity was assessed every 2 weeks. RESULTS: Twenty-one patients were treated on study. The most frequently reported treatment-related grade 2/3 adverse events were hand-foot syndrome (29%), leukopenia/neutropenia (24%), and fatigue (19%). Grade 3 toxicity was transient and easily managed. Three patients experienced grade 3 hand-foot syndrome; one of these patients had grade 3 diarrhea. There were no grade 4 events. The MTD of capecitabine 7/7 is 2,000 mg twice daily. CONCLUSION: As predicted by mathematical modeling, capecitabine dosing for 7 days followed by a 7-day rest is well tolerated. Efficacy of this schedule is being determined in a phase II clinical trial in patients with advanced breast cancer. PMID- 18398146 TI - Variation in bleomycin hydrolase gene is associated with reduced survival after chemotherapy for testicular germ cell cancer. AB - PURPOSE: Response to chemotherapy may be determined by gene polymorphisms involved in metabolism of cytotoxic drugs. A plausible candidate is the gene for bleomycin hydrolase (BLMH), an enzyme that inactivates bleomycin, an essential component of chemotherapy regimens for disseminated testicular germ-cell cancer (TC). We investigated whether the single nucleotide polymorphism (SNP) A1450G of the BLMH gene (rs1050565) is associated with survival. PATIENTS AND METHODS: Data were collected on survival and BLMH genotype of 304 patients with TC treated with bleomycin-containing chemotherapy at the University Medical Center Groningen, the Netherlands, between 1977 and 2003. Survival according to genotype was analyzed using Kaplan-Meier curves with log-rank testing and Cox regression analysis with adjustment for confounders. RESULTS: BLMH gene SNP A1450G has a significant effect on TC-related survival (log-rank P = .001). The homozygous variant (G/G) genotype (n = 31) is associated with decreased TC related survival compared with the heterozygous variant (A/G; n = 133) and the wild-type (A/A; n = 140). With Cox regression the G/G genotype proves to be an unfavorable prognostic factor, in addition to the commonly used International Germ Cell Consensus Classification prognosis group, with a hazard ratio of 4.97 (95% CI, 2.17 to 11.39) for TC related death. Furthermore, the G/G genotype shows a higher prevalence of early relapses. CONCLUSION: The homozygous variant G/G of BLMH gene SNP A1450G is associated with reduced survival and higher prevalence of early relapses in TC patients treated with bleomycin-containing chemotherapy. This association is hypothesis generating and may eventually be of value for risk classification and selection for alternative treatment strategies in patients with disseminated TC. PMID- 18398147 TI - Toremifene improves lipid profiles in men receiving androgen-deprivation therapy for prostate cancer: interim analysis of a multicenter phase III study. AB - PURPOSE: Androgen-deprivation therapy (ADT) is associated with greater risk of incident coronary heart disease and hospital admission for myocardial infarction; treatment-related increases in serum lipids may contribute to greater cardiovascular disease risk. We evaluated the effects of toremifene, a selective estrogen-receptor modulator, on fasting serum lipid levels in men receiving ADT for prostate cancer. PATIENTS AND METHODS: In an ongoing, multicenter, double blind, placebo-controlled phase III fracture-prevention study, 1,389 men receiving ADT for prostate cancer were randomly assigned to receive toremifene (80 mg/d) or placebo. In this interim analysis of 188 patients, changes in fasting serum lipids from baseline to month 12 were compared between the placebo and toremifene groups. RESULTS: Changes in serum lipids differed significantly between the groups. Mean (+/- SE) total cholesterol decreased by 1.0% +/- 1.7% from baseline to month 12 in the placebo group and decreased by 8.1% +/- 1.4% in the toremifene group (P = .001 for between group comparison). Low-density lipoprotein (LDL) cholesterol increased by 0.8% +/- 2.5% in the placebo group and decreased by 8.2% +/- 2.5% in the toremifene group (P = .003). In contrast, high density lipoprotein (HDL) cholesterol decreased by 4.9% +/- 1.2% in the placebo group and increased by 0.5% +/- 2.2% in the toremifene group (P = .018). Triglycerides increased by 6.9% +/- 4.2% in the placebo group and decreased by 13.2% +/- 3.6% in the toremifene group (P = .003). CONCLUSION: Toremifene significantly decreased total cholesterol, LDL cholesterol, and triglycerides, and increased HDL cholesterol in men receiving ADT for prostate cancer. PMID- 18398148 TI - Bevacizumab, capecitabine, and oxaliplatin as neoadjuvant therapy for patients with potentially curable metastatic colorectal cancer. AB - PURPOSE: Patients with colorectal cancer (CRC) and liver metastases have a poor prognosis, but can benefit from perioperative chemotherapy and disease resection. Bevacizumab improves outcomes in patients with metastatic CRC; however, its impact on surgical complications and hepatic regeneration after liver resection remains to be determined. PATIENTS AND METHODS: Fifty-six patients with metastatic CRC with liver metastases potentially curable by resection were eligible for this single-center, nonrandomized phase II trial. Eligibility criteria defined patients at high risk of early recurrence. Patients received biweekly bevacizumab plus capecitabine and oxaliplatin for six cycles. The sixth cycle of therapy did not include bevacizumab, resulting in 5 weeks between the last administration of bevacizumab and surgery. RESULTS: Objective response to neoadjuvant chemotherapy was achieved in 41 patients (73%). Fifty-two patients underwent liver resection including 11 with synchronous primary tumor resection. No increased intraoperative bleeding events or wound-healing complications were observed and only three patients (6%) required perioperative blood transfusions. Further surgery was necessary in a single patient. Postoperative liver function and regeneration were normal in all but one patient. No postoperative mortality occurred and morbidity was encountered in 11 patients (20%). The mean length of postoperative hospitalization was 9 days (+/- 4.0). CONCLUSION: These data suggest that bevacizumab can be safely administered until 5 weeks before liver resection in patients with metastatic CRC without increasing the rate of surgical or wound healing complications or severity of bleeding. To our knowledge, they are also the first to show that neoadjuvant bevacizumab does not affect liver regeneration after resection. PMID- 18398149 TI - Effect of molecular therapeutics on liver regeneration in a murine model. AB - PURPOSE: Unresectable metastatic colorectal cancer (CRC) can be rendered resectable with systemic chemotherapy in approximately 20% of cases. Most patients with metastatic CRC receive chemotherapy with the addition of targeted therapy with anti-vascular endothelial growth factor (VEGF) or anti-epidermal growth factor receptor (EGFR) antibodies. We sought to determine whether anti VEGF receptor (VEGFR) or anti-EGFR therapy would impair liver regeneration after partial hepatectomy (PH) in mice. MATERIALS AND METHODS: Mice underwent either 66% PH or sham laparotomy. In the first experiment, mice in the PH group were randomly assigned to receive daily intraperitoneal injections of monoclonal antibodies (MoABs) to murine VEGFR-2 or nonspecific MoABs (control). In the second experiment, mice in the PH group were randomly assigned to receive intraperitoneal injections of antimurine EGFR or nonspecific (control) MoABs. In both experiments, therapy was initiated the day before surgery and continued until the mice were killed on day 5. Livers were collected and processed. RESULTS: Anti-VEGFR-2 therapy slightly impaired liver regeneration and hepatic cell proliferation compared with control. Hematoxylin and eosin staining showed no differences in liver morphology. CD105 staining showed decreased levels of activated endothelium in livers in the VEGFR-2 MoAB group. VEGFR-2 MoAB therapy decreased the levels of the cell cycle regulators cyclin D1 and cyclin D3 and the regenerative cytokine interleukin-6. Anti-EGFR therapy had no effect on liver regeneration or cellular proliferation. CONCLUSION: Anti-VEGFR-2 therapy slightly impaired liver regeneration in this murine model, whereas anti-EGFR therapy had no effect on liver regeneration. PMID- 18398150 TI - Comparison of the results of the treatment of adolescents and young adults with standard-risk acute lymphoblastic leukemia with the Programa Espanol de Tratamiento en Hematologia pediatric-based protocol ALL-96. AB - PURPOSE: Retrospective studies have shown that adolescents and young adults with acute lymphoblastic leukemia (ALL) treated with pediatric protocols have better outcomes than similarly aged patients treated with adult protocols, but prospective studies comparing adolescents and young adults using pediatric schedules are scarce. The ALL-96 protocol was addressed to compare the toxicity and results of a pediatric-based protocol in adolescents (age 15-18 years) and young adults (age 19-30 years) with standard-risk (SR) ALL. PATIENTS AND METHODS: Adolescents (n = 35) and young adults (n = 46) received a standard five-drug/5 week induction course followed by two cycles of early consolidation, maintenance with monthly reinforcement cycles up to 1 year in continuous complete remission (CR) and 1 year with standard maintenance chemotherapy up to 2 years in CR. RESULTS: Adolescents and young adults were comparable in the main pretreatment ALL characteristics. The CR rate was 98% and. after a median follow-up of 4.2 years, 6-year event-free survival (EFS) and overall survival (OS) were 61% (95% CI, 51% to 72%) and 69% (95% CI, 59% to 79%), respectively, with no differences between adolescents and young adults. The hematologic toxicity in consolidation and reinforcement cycles was higher in young adults than in adolescents. Slow response to induction therapy was the only parameter associated with poor EFS (34% v 67%) and OS (40% v 76%). CONCLUSION: The response to the pediatric ALL-96 protocol was identical in adolescents and young adults despite a slight increase in hematologic toxicity observed in adults. This justifies the age-unrestricted use of pediatric regimens to treat patients with SR ALL. PMID- 18398151 TI - Lack of community insurance and mammography screening rates among insured and uninsured women. AB - PURPOSE: To evaluate whether the proportion of the local population without health insurance coverage is related to whether women undergo mammography screening. METHODS: Survey data on 12,595 women 40 to 69 years of age from the 2000 to 2001 Community Tracking Study Household Survey were used to analyze the relation between community lack of insurance and whether the respondent had a mammogram within the past year. RESULTS: Women age 40 to 69 were less likely to report that they had a mammogram within the last year if they resided in communities with a relatively high uninsurance rate, even after adjusting for other factors. After adjusting for individual insurance and other factors, a 10 percentage-point decrease in the proportion of the local insured population is associated with a 17% (95% CI, 13% to 21%) decrease in the odds that a woman age 40 to 69 years will undergo mammography screening within a year. CONCLUSION: Women living in communities with high uninsurance are substantially less likely to undergo mammography screening. These results are consistent with the view that the negative impact of uninsurance extends to everyone in the community regardless of individual health insurance status. PMID- 18398152 TI - Phase I and pharmacokinetic study of daily oral AZD2171, an inhibitor of vascular endothelial growth factor tyrosine kinases, in combination with carboplatin and paclitaxel in patients with advanced non-small-cell lung cancer: the National Cancer Institute of Canada clinical trials group. AB - PURPOSE: AZD2171 is a potent inhibitor of vascular endothelial growth factor receptors that showed broad antitumor activity in preclinical models. Doses of up to 45 mg/d of AZD2171 are tolerable when administered alone. This study evaluated escalating doses of AZD2171 in combination with standard chemotherapy in patients with advanced non-small-cell lung cancer. PATIENTS AND METHODS: Eligible patients received carboplatin targeted to an area under the concentration time curve of 6 mg . min/mL and paclitaxel 200 mg/m(2), both on day 1 of a 3-week cycle; daily oral AZD2171 at either 30 mg or 45 mg commenced day 2 of cycle 1. Pharmacokinetics of all drugs were performed, and tumor response was assessed by Response Evaluation Criteria in Solid Tumors (RECIST). RESULTS: Twenty patients were enrolled. No dose-limiting toxicities were observed during cycle 1 at either dose. Fatigue, diarrhea, anorexia, and granulocytopenia were common; hypertension was manageable with a treatment algorithm designed for this protocol. No clinically significant drug-related bleeding was observed. At 45 mg/d, fatigue and diarrhea were increased, and headache and hoarseness were observed. Paclitaxel clearance decreased during cycle 2, but no other significant pharmacokinetic interactions were observed. After radiology review, confirmed responses were observed in nine patients (response rate, 45%; 95% CI, 23% to 68%); all but one enrolled patient showed evidence of tumor shrinkage, some with cavitation. CONCLUSION: AZD2171 can be combined with standard doses of carboplatin/paclitaxel with encouraging antitumor activity. Toxicity is increased, but predictable and manageable. PMID- 18398153 TI - Phase III trial comparing carboplatin, paclitaxel, and bexarotene with carboplatin and paclitaxel in chemotherapy-naive patients with advanced or metastatic non-small-cell lung cancer: SPIRIT II. AB - PURPOSE: The purpose of this study was to determine whether addition of the synthetic rexinoid bexarotene (Targretin; Eisai Inc, Woodcliff Lake, NJ) to standard first-line carboplatin and paclitaxel therapy provides additional survival benefit in patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients with stage IIIB disease with pleural effusion, or stage IV NSCLC and Eastern Cooperative Oncology Group performance status 0 to 1, were randomly assigned to bexarotene 400 mg/m(2)/d combined with carboplatin and paclitaxel, or assigned to carboplatin and paclitaxel alone. Bexarotene patients also received lipid-lowering agents on or before day 1. The primary efficacy end point was overall survival; secondary efficacy and supportive analyses were also conducted. RESULTS: A total of 612 patients (306 per arm) were enrolled onto the study. In the intent-to-treat population, no significant difference in survival occurred between the two arms. However, a subpopulation (approximately 40%) of bexarotene-treated patients who experienced National Cancer Institute grade 3/4 hypertriglyceridemia had significantly longer median survival than control patients (12.4 v 9.2 months; log-rank, P = .014). Bexarotene-treated patients with grade 3/4 hypertriglyceridemia who received the most benefit included those who were male, were smokers, experienced 6-month prior weight loss >or= 5%, and had stage IV disease. The incidence and severity of most adverse events were similar between arms, although hyperlipidemia, neutropenia, fatigue, leukopenia, arthralgia, and diarrhea were more frequent in the bexarotene arm. CONCLUSION: Although the addition of bexarotene to chemotherapy did not improve survival in the overall study population, occurrence of high-grade hypertriglyceridemia in bexarotene-treated patients strongly correlated with increased survival, suggesting that bexarotene may benefit a segment of first-line NSCLC patients. PMID- 18398154 TI - Randomized phase III trial comparing bexarotene (L1069-49)/cisplatin/vinorelbine with cisplatin/vinorelbine in chemotherapy-naive patients with advanced or metastatic non-small-cell lung cancer: SPIRIT I. AB - PURPOSE: This study evaluated whether the combination of the synthetic rexinoid bexarotene with first-line cisplatin/vinorelbine therapy provides additional survival benefit in patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients with stage IIIB with pleural effusion or stage IV NSCLC and Eastern Cooperative Oncology Group performance status 0 to 1 were randomly assigned to open-label bexarotene 400 mg/m(2)/d with cisplatin/vinorelbine or to cisplatin/vinorelbine alone. Antilipid agents were initiated on or before day 1 in the bexarotene arm. Primary efficacy end point was overall survival. Primary, secondary and supportive efficacy analyses were conducted. RESULTS: A total of 623 patients (312 control, 311 bexarotene) were enrolled. Overall, no significant difference in survival occurred between the two treatment groups. However, an unplanned retrospective analysis showed that a subpopulation of bexarotene patients (n = 98 of 306) who experienced National Cancer Institute grade 3/4 hypertriglyceridemia had longer median survival compared with control patients (12.3 v 9.9 months; log-rank P = .08). Within that subgroup, those who benefited the most included males, smokers, those with stage IV disease, and those with a 6-month prior weight loss of 5% or more. Incidence, type and severity of grade 3/4 adverse events were comparable between arms, except for leukopenia (higher in chemotherapy arm) and hyperlipemia, hypothyroidism, dyspnea, and headache (higher in chemotherapy/bexarotene arm). CONCLUSION: The addition of bexarotene to first-line chemotherapy did not increase survival in patients with advanced NSCLC. However, a subgroup (32%) of bexarotene-treated patients developing high-grade hypertriglyceridemia appeared to have better survival (12.3 months) than controls; thus triglyceride response may be a biomarker of survival benefit with bexarotene. PMID- 18398155 TI - Improvement in oncology practice performance through voluntary participation in the Quality Oncology Practice Initiative. AB - PURPOSE: The Quality Oncology Practice Initiative (QOPI) became available to all American Society of Clinical Oncology member physicians in 2006 as a voluntary medical oncology practice-based quality measurement and improvement project. QOPI assesses practice performance for a series of evidence- and consensus-based process measures, relying on practices to complete structured chart reviews and submit data via a secure Web-based portal. METHODS: This analysis focused on the 71 practices that participated in both the March and September 2006 data collections (7,624 charts abstracted in March and 10,240 in September). Among 33 measures common to both collections, five measures were closely correlated, and 28 are included in the final analysis. Composite scores were created for six different domains of care. Statistical significance was tested on both absolute changes and relative changes (relative failure reduction) of quality measures from baseline to follow-up and between the lower quartile and all other quartiles. RESULTS: Practice performance on individual measures varied between 18.8% and 98.6%. Mean overall performance as measured by a composite score increased from 78.7% in March to 82.3% in September (P < .05). Improvement was most marked among practices originally performing in the bottom quartile. Using a composite score, the absolute and relative performance for the bottom quartile improved by 27% and 35%, respectively, statistically superior to that of all others. CONCLUSION: Practices that participated in QOPI demonstrated improved performance in self-reported process measures, with the greatest improvement demonstrated in initially low-performing practices. PMID- 18398156 TI - Compassionate approval process for experimental gene-based products. PMID- 18398157 TI - Could the efficacy of docetaxel in prostate cancer patients be potentiated by concomitant high-dose calcitriol administration? PMID- 18398158 TI - No evidence for taxane/platinum pharmacogenetic markers: just lack of power? PMID- 18398159 TI - Treatment of venous thrombosis in patients with intracranial malignancies. PMID- 18398160 TI - Low platelet counts in cancer patients: should heparin for venous thromboembolism prophylaxis be instituted? PMID- 18398161 TI - Capecitabine and irinotecan as first-line treatment of advanced colorectal cancer. PMID- 18398162 TI - Contralateral prophylactic mastectomy: mind the genetics. PMID- 18398164 TI - 16S rRNA-, GroEL- and MucZ-based assessment of the taxonomic position of 'Rickettsiella melolonthae' and its implications for the organization of the genus Rickettsiella. AB - 'Rickettsiella melolonthae' is an intracellularly multiplying bacterial pathogen of European cockchafers, Melolontha melolontha (Linnaeus, 1758) and Melolontha hippocastani (Fabricius, 1801) (Coleoptera: Scarabaeidae). We report the first determination of nucleotide sequences from this organism, i.e. the 16S rRNA encoding rrs gene, the chaperonin encoding groEL gene and the mucZ gene encoding the orthologue of a capsule synthesis-inducing factor of Coxiella burnetii. Within the genus Rickettsiella, the pathotype 'Rickettsiella melolonthae' is currently classified as a synonym of the nomenclatural type species Rickettsiella popilliae. Previous sequencing of a 16S rRNA gene from a different species, Rickettsiella grylli, has motivated the transfer of the entire genus from the alphaproteobacterial order Rickettsiales to the gammaproteobacterial order Legionellales, family Coxiellaceae. We investigated the validity of this taxonomic reorganization beyond the species Rickettsiella grylli by reconstructing the organismal phylogeny from comparisons of 16S rRNA gene and GroEL and MucZ protein sequences from a selected set of alpha- and gammaproteobacteria as well as bacterial pathogens from the order Chlamydiales. Our analysis strongly supported the transfer of the genus Rickettsiella to the order Legionellales, but not its classification in one of the recognized families present in this order. Furthermore, our results substantiated inconsistencies in the internal organization of the genus. In particular, the currently accepted delineation of Rickettsiella species and the claimed synonymy of 'Rickettsiella melolonthae' with Rickettsiella popilliae are not simultaneously consistent with our findings. PMID- 18398165 TI - Flavobacterium anhuiense sp. nov., isolated from field soil. AB - A novel strain, D3T, isolated from a field-soil sample obtained from Anhui Province, PR China, was characterized taxonomically by using a polyphasic approach. The cells were Gram-negative, yellow-pigmented rods devoid of flagella, but showing gliding motility. The organism was able to grow at 5-37 degrees C and at pH 4.0-10.0. A comparative 16S rRNA gene sequence analysis indicated that strain D3T is a member of the genus Flavobacterium, sharing highest sequence similarity with the type strain of Flavobacterium defluvii (96.7 %). The major isoprenoid quinone was MK-6 and the predominant fatty acids were iso-C15 : 0, summed feature 3 (C16 : 1 omega 7c and/or iso-C15 : 0 2-OH) and C16 : 0. The DNA G+C content was 31.4 mol%. On the basis of phylogenetic and phenotypic data, strain D3T represents a novel species within the genus Flavobacterium, for which the name Flavobacterium anhuiense sp. nov. is proposed. The type strain is D3T (=KCTC 22128T = CGMCC 1.6859T). PMID- 18398166 TI - Tenacibaculum aiptasiae sp. nov., isolated from a sea anemone Aiptasia pulchella. AB - A novel bacterial strain, designated a4T, isolated from a sea anemone (Aiptasia pulchella) in Taiwan, was characterized using a polyphasic taxonomic approach. Strain a4T was aerobic, Gram-negative, pale-yellow-pigmented and rod-shaped. It grew optimally at 30-35 degrees C, in the presence of 3-4 % (w/v) NaCl and at pH 8.0. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain belonged to the genus Tenacibaculum (family Flavobacteriaceae, phylum Bacteroidetes). The closest neighbours were Tenacibaculum lutimaris TF-26T (97.6 % similarity) and Tenacibaculum aestuarii SMK-4T (97.7 % similarity). The novel isolate could be distinguished from all Tenacibaculum species by several phenotypic characteristics. The major fatty acids were summed feature 3 (comprising C16 : 1 omega 7c and/or iso-C15 : 0 2-OH, 19.6 %), iso-C15 : 0 (12.9 %), iso-C16 : 0 3-OH (10.2 %), iso-C17 : 0 3-OH (9.9 %) and iso-C15 : 1 (9.5 %). The DNA G+C content was 35.0 mol%. Hence, genotypic and phenotypic data demonstrate that strain a4(T) should be classified as a representative of a novel species in the genus Tenacibaculum, for which the name Tenacibaculum aiptasiae sp. nov. is proposed. The type strain is a4T (=BCRC 17655T =LMG 24004T). PMID- 18398167 TI - Proposal to reclassify the three biotypes of Bifidobacterium longum as three subspecies: Bifidobacterium longum subsp. longum subsp. nov., Bifidobacterium longum subsp. infantis comb. nov. and Bifidobacterium longum subsp. suis comb. nov. AB - In the year 2002, Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium suis were unified into a single species, Bifidobacterium longum, preserving the former species names through the creation of the three biotypes 'longum', 'infantis' and 'suis'. Consequently, the use of the species names B. infantis and B. suis was to be discontinued. The above taxonomic rearrangement of B. longum was based on DNA-DNA hybridizations and 16S rRNA and HSP60 gene sequence analysis. However, a variety of other genotypic techniques including ribotyping, amplified rDNA restriction analysis (ARDRA), randomly amplified polymorphic DNA (RAPD)-PCR, BOX-PCR, PCR-denaturing gradient gel electrophoresis (DGGE), comparison of the recA, tuf and ldh gene sequences, plasmid profiling and considerable variation in carbohydrate fermentation patterns as well as results of starch and PAGE electrophoresis experiments clearly discriminate former B. longum, B. infantis and B. suis strains. In the present paper we compile this published information and propose the description of Bifidobacterium longum subsp. longum subsp. nov., Bifidobacterium longum subsp. infantis comb. nov. and Bifidobacterium longum subsp. suis comb. nov. The International Committee on Systematics of Prokaryotes Subcommittee on the taxonomy of Bifidobacterium, Lactobacillus and related organisms is in favour of this proposal. The type strains of Bifidobacterium longum subsp. longum subsp. nov., subsp. infantis comb. nov. and subsp. suis comb. nov. are E194b (variant a)T (ATCC 15707T=DSM 20219T), S12T (=ATCC 15697T=DSM 20088T) and Su859T (ATCC 27533T=DSM 20211T), respectively. PMID- 18398168 TI - Elioraea tepidiphila gen. nov., sp. nov., a slightly thermophilic member of the Alphaproteobacteria. AB - A bacterial isolate, strain TU-7T, with an optimum growth temperature of about 45 50 degrees C and an optimum pH for growth between 8.0 and 8.5, was recovered from a hot spring in the Furnas area of the island of Sao Miguel in the Azores. The new isolate is non-pigmented, facultatively chemolithoorganotrophic, strictly aerobic and catalase- and oxidase-positive. The organism oxidized thiosulfate to sulfate with enhancement of growth. This organism assimilated organic acids but did not assimilate carbohydrates or polyols. 16S rRNA gene sequence analysis placed strain TU-7T within the radiation of the Alphaproteobacteria as a deep branch of the family Acetobacteriaceae. The major fatty acids of strain TU-7T are 18 : 1 omega 7c, 18 : 0, 19 : 0 cyclo omega 8c and an aliphatic chain with an equivalent chain-length of 20.195, identified as 19 : 0 2-OH cyclo omega 8 (Delta 11 : 12). Ubiquinone 10 is the major respiratory quinone and the major polar lipids are phosphatidylcholine, phosphatidylethanolamine, diphosphosphatidylglycerol and phosphatidylglycerol in addition to two unidentified aminolipids. Bacteriochlorophyll a and puf genes were not detected. Based on 16S rRNA gene sequence analysis and physiological and biochemical characteristics, we describe a novel species of a new genus represented by strain TU-7T, for which we propose the name Elioraea tepidiphila gen. nov., sp. nov. The type strain of Elioraea tepidiphila is strain TU-7T (=DSM 17972T =CIP 109115T). PMID- 18398169 TI - Yersinia massiliensis sp. nov., isolated from fresh water. AB - Two bacterial organisms, 50640T and 823, were isolated from fresh water in Marseilles, France, and were further identified as members of the genus Yersinia on the basis of their phenotypic characteristics and 16S rRNA gene sequencing. Their unique phenotypic profile differed from that of closely related species of Yersinia bercovieri and Yersinia mollaretii by exhibiting positive indole and inositol tests, and from that of Yersinia frederiksenii by lacking the ability to ferment l-rhamnose. A polyphasic approach, including almost complete 16S rRNA gene sequencing (1461 bp) and partial sequencing of hsp60 (683 bp), gyrB (662 bp), sodA (624 bp) and rpoB (1049 bp) showed that isolates 50640T and 823 exhibited 98.5, 93.5, 90.4, 92.4 and 96.6 % similarity with Y. mollaretii, 98.7, 93.0, 90.1, 89.1 and 96.2 % with Y. bercovieri, and 98.4, 93.2, 89.8, 88.9 and 95.2 % with Y. frederiksenii, respectively. Both isolates exhibited an identical 16S rRNA gene sequence and differed by one to five point mutations in housekeeping gene sequences. Phylogenetic reconstructions based on the combination of these four housekeeping genes indicated that the two isolates formed a unique branch supported by a bootstrap value of 93 %. Their unique phenotypic traits, 16S rRNA gene sequence, together with housekeeping gene sequences exhibiting <97 % similarity with closely related species, and phylogenetic analyses suggested that the two isolates represent a so far undescribed Yersinia species. The name Yersinia massiliensis sp. nov. is proposed for this new taxon (type strain 50640T=CIP 109351T=CCUG 53443T; isolate 823=CIP 109352=CCUG 53444). PMID- 18398170 TI - Tessaracoccus flavescens sp. nov., isolated from marine sediment. AB - A novel actinobacterium, designated strain SST-39T, was isolated from a marine sediment sample collected in Jeju, Republic of Korea. Cells were facultatively anaerobic, Gram-positive, non-endospore-forming, non-motile rods. Colonies were circular, slightly convex, opaque and brilliant yellow. The cell-wall peptidoglycan of the organism contained ll-diaminopimelic acid as the diagnostic diamino acid. Polar lipids included phosphatidylglycerol and diphosphatidylglycerol. The predominant menaquinone was MK-9(H4). The major cellular fatty acids were anteiso-C15 : 0, C18 : 0 and C16 : 0. The DNA G+C content was 68.4 mol%. A phylogenetic tree based on 16S rRNA gene sequences indicated that the isolate belonged to the family Propionibacteriaceae and formed a unique cluster with the type strain of Tessaracoccus bendigoensis (97.0 % sequence similarity). Other phylogenetic neighbours were the type strains of Luteococcus peritonei (95.1 % 16S rRNA gene sequence similarity) and Propionibacterium propionicum (95.1 %). On the basis of its phenotypic and phylogenetic distinctiveness, strain SST-39T is considered to represent a novel species of the genus Tessaracoccus, for which the name Tessaracoccus flavescens sp. nov. is proposed. The type strain is SST-39T (=DSM 18582T =KCTC 19196T). PMID- 18398171 TI - Description of Maribacter forsetii sp. nov., a marine Flavobacteriaceae isolated from North Sea water, and emended description of the genus Maribacter. AB - Three rod-shaped, Gram-negative, chemo-organotrophic, heterotrophic, strictly aerobic, gliding bacterial strains, KT02ds18-4, KT02ds18-5 and KT02ds18-6T, were isolated from North Sea surface waters near the island of Helgoland, Germany. Their taxonomic position was investigated by a polyphasic approach. The three strains were light yellow, oxidase- and catalase-positive, and grew optimally at 25 degrees C, at pH 7.5, and in the presence of 2.5 % (w/v) NaCl. The Chargaff's coefficient was 34.2-34.4 mol%. The three strains shared >90 % DNA-DNA relatedness and an identical 16S rRNA gene sequence. Comparative 16S rRNA gene sequence analysis allocated the three strains to the genus Maribacter in the family Flavobacteriaceae, with similarities of 97.0-97.4 % to five of the recognized Maribacter species. Their low level of DNA-DNA relatedness (<20 %) with these species and differentiating phenotypic characteristics demonstrated that they constitute a new Maribacter species for which the name Maribacter forsetii sp. nov. is proposed. Strain KT02ds18-6T (=CIP 109504T=DSM 18668T) is the type strain. An emended description of the genus Maribacter is also proposed. PMID- 18398172 TI - Haloferax elongans sp. nov. and Haloferax mucosum sp. nov., isolated from microbial mats from Hamelin Pool, Shark Bay, Australia. AB - Extremely halophilic archaea were cultivated from smooth and pustular microbial mats collected from Hamelin Pool, Shark Bay, Western Australia. On the basis of morphology, two phenotypes were present and 16S rRNA gene sequence analysis indicated that all strains were most closely related to members of the genus Haloferax (98.1-99.4 % similarity). One representative strain from each phenotype was selected for further taxonomic characterization. Strain SA5T, isolated from the smooth mat, formed small ( approximately 1 mm diameter), red, translucent colonies on agar medium and strain PA12T, isolated from the pustular mat, formed large (3-5 mm diameter), pink, mucoid, domed colonies. Both strains grew in media with 1.7-5.1 M NaCl, required at least 0.2 M Mg2+ for growth and had pH optima of 7.4. The 16S rRNA gene similarity between strains SA5T and PA12T was 97.1 %. Physiological properties, G+C content and polar lipid composition supported placement of both strains in the genus Haloferax. Phenotypic analysis indicated that the two strains were distinct from each other and from all other members of the genus. This was confirmed by the low DNA-DNA relatedness between strains SA5T and PA12T (18-30 %) and between both strains and all other recognized Haloferax species. Two novel species of the genus Haloferax are proposed to accommodate these novel isolates, Haloferax elongans sp. nov. (type strain SA5T=JCM 14791T=ATCC BAA-1513T=UNSW 104100T) and Haloferax mucosum sp. nov. (type strain PA12T=JCM 14792T=ATCC BAA-1512T=UNSW 104200T). PMID- 18398173 TI - Halomonas cerina sp. nov., a moderately halophilic, denitrifying, exopolysaccharide-producing bacterium. AB - Three bacterial strains were isolated from different saline soils in Spain. The novel strains were moderately halophilic, exopolysaccharide-producing, Gram negative, non-motile rods. The strains required NaCl and grew best with 7.5-10 % (w/v) NaCl in the medium. They formed wax-coloured colonies, were oxidase positive and showed respiratory metabolism, using oxygen, nitrate and nitrite as terminal electron acceptors. The novel strains were able to denitrify and did not produce acid from sugars. The DNA G+C contents varied between 62.7 and 66.2 mol%. Phylogenetic analyses based on 16S rRNA gene sequences and sequence signatures of this gene showed that all three novel isolates belonged to the genus Halomonas in the class Gammaproteobacteria and formed an independent phylogenetic line. The most phylogenetically related species were Halomonas alimentaria, Halomonas campaniensis, Halomonas gudaonensis and Halomonas ventosae, with which the novel strains showed 16S rRNA gene sequence similarity values of between 96.3 and 95.2 %. The principal fatty acids of the novel strains were 16 : 0, 18 : 1 omega 7c, 16 : 1 omega 7c and 19 : 0 cyclo omega 8c. The predominant respiratory lipoquinone was ubiquinone with nine isoprene units (Q-9). The name Halomonas cerina sp. nov. is proposed for these isolates. The type strain is SP4T (=CECT 7282T=LMG 24145T). PMID- 18398174 TI - Archaeoglobus infectus sp. nov., a novel thermophilic, chemolithoheterotrophic archaeon isolated from a deep-sea rock collected at Suiyo Seamount, Izu-Bonin Arc, western Pacific Ocean. AB - A novel thermophilic, strictly anaerobic archaeon, designated strain Arc51T, was isolated from a rock sample collected from a deep-sea hydrothermal field in Suiyo Seamount, Izu-Bonin Arc, western Pacific Ocean. Cells of the isolate were irregular cocci with single flagella and exhibited blue-green fluorescence at 436 nm. The optimum temperature, pH and NaCl concentration for growth were 70 degrees C, pH 6.5 and 3 % (w/v), respectively. Strain Arc51T could grow on thiosulfate or sulfite as an electron acceptor in the presence of hydrogen. This strain required acetate as a carbon source for its growth, suggesting that the reductive acetyl CoA pathway for CO2 fixation was incomplete. In addition, coenzyme M (2 mercaptoethanesulfonic acid), which is a known methyl carrier in methanogenesis, was also a requirement for growth of the strain. Analysis of the 16S rRNA gene sequence revealed that the isolate was similar to members of the genus Archaeoglobus, with sequence similarities of 93.6-97.2 %; the closest relative was Archaeoglobus veneficus. Phylogenetic analyses of the dsrAB and apsA genes, encoding the alpha and beta subunits of dissimilatory sulfite reductase and the alpha subunit of adenosine-5'-phosphosulfate reductase, respectively, produced results similar to those inferred from comparisons based on the 16S rRNA gene sequence. On the basis of phenotypic and phylogenetic data, strain Arc51T represents a novel species of the genus Archaeoglobus, for which the name Archaeoglobus infectus sp. nov. is proposed. The type strain is Arc51T (=NBRC 100649T=DSM 18877T). PMID- 18398175 TI - Moritella dasanensis sp. nov., a psychrophilic bacterium isolated from the Arctic ocean. AB - An aerobic, motile, Gram-negative, ice-active substance-producing, rod-shaped psychrophile, designated strain ArB 0140T, was isolated from seawater collected from near a glacier in Kongsfjorden, Svalbard Archipelago, Norway. Phylogenetic analysis using 16S rRNA gene sequences indicated that strain ArB 0140T showed a distinct phyletic line within the genus Moritella. Characteristic chemotaxonomic data [predominant isoprenoid quinone, Q8; major fatty acids, C14 : 0, C14 : 1, C16 : 0, C16 : 1 and C22 : 6 (docosahexaenoic acid; DHA)] also corroborated the affiliation of strain ArB 0140T to the genus Moritella. The maximal growth rate of the novel strain was observed at 9 degrees C, with a maximum temperature for growth of 18 degrees C. The genomic DNA G+C content was 46.9 mol%. Based on the data obtained from this polyphasic study, including DNA-DNA relatedness, physiological and biochemical tests and ice-controlling activity, strain ArB 0140T was found to be genetically and phenotypically different from other recognized species of the genus Moritella. Therefore strain ArB 0140T represents a novel species, for which the name Moritella dasanensis sp. nov. is proposed. The type strain is ArB 0140T (=KCTC 10814T=KCCM 42845T=JCM 14759T). PMID- 18398176 TI - Desulfovibrio psychrotolerans sp. nov., a psychrotolerant and moderately alkaliphilic sulfate-reducing deltaproteobacterium from the Himalayas. AB - A novel sulfate-reducing bacterium (strain JS1T) was isolated from lake Pangong, a salt-water lake situated in the Himalayas of Leh District, Jammu and Kashmir, India. Cells of strain JS1T are Gram-negative, vibriod, motile by means of single polar flagellum, contain desulfoviridin, are catalase-positive and can hydrolyse starch. Strain JS1T grew optimally at pH 8.5-9.0 and at temperatures ranging from 10 to 50 degrees C (optimum 28-30 degrees C). Chemo-organoheterotrophy was the only growth mode of strain JS1T and was possible on a wide range of organic compounds. Sulfate alone but not sulfite, thiosulfate, elemental sulfur, nitrite or nitrate was used as electron acceptor. It has no requirement for NaCl but can tolerate up to 2 % NaCl (w/v). p-Aminobenzoate is a required growth factor. The 16S rRNA gene sequence of the bacterium demonstrated similarity between strain JS1T and the members of the genus Desulfovibrio. The position of strain JS1T within the phylogenetic tree indicated that it clustered closely with Desulfovibrio acrylicus DSM 10141(T) (89.85 % sequence similarity). On the basis of phenotypic and molecular genetic evidence, it is proposed that strain JS1T be classified as a novel species of the genus Desulfovibrio of the family Desulfovibrionaceae, with the species name Desulfovibrio psychrotolerans sp. nov. The type strain of the species is JS1T (=JCM 14597T=KCTC 5573T=DSM 19430T). PMID- 18398177 TI - Desulfoluna butyratoxydans gen. nov., sp. nov., a novel Gram-negative, butyrate oxidizing, sulfate-reducing bacterium isolated from an estuarine sediment in Japan. AB - A strictly anaerobic, mesophilic, sulfate-reducing bacterial strain, designated MSL71T, was isolated from an estuarine sediment from the Sea of Japan bordering the Japanese islands and was characterized phenotypically and phylogenetically. The cells were found to be Gram-negative, motile, non-spore-forming, slightly curved rods. Catalase and oxidase activities were not detected. The optimum NaCl concentration for growth was 2.0 % (w/v), the optimum temperature was 30 degrees C and the optimum pH was 6.3. Strain MSL71T utilized formate, butyrate, pyruvate, lactate, malate, ethanol, propanol, butanol, glycerol and H2 as electron donors for sulfate reduction. The organic electron donors used were incompletely oxidized, mainly to acetate. The strain did not use acetate, propionate, fumarate, succinate, methanol, glycine, alanine, serine, aspartate or glutamate. Sulfite and thiosulfate were used as electron acceptors with lactate as an electron donor, but fumarate was not utilized. Without electron acceptors, pyruvate and malate, but not lactate or fumarate, were fermented. The genomic DNA G+C content was 62.0 mol%. Menaquinone MK-8(H4) was the major respiratory quinone. The major cellular fatty acids were C14 : 0, C16 : 0, C16 : 1 omega 7, C18 : 1 omega 9, C18 : 1 omega 7 and C14 : 0 3-OH. A phylogenetic analysis based on the 16S rRNA gene sequence placed the strain in the class Deltaproteobacteria. The closest recognized relative of strain MSL71T was Desulfofrigus fragile (93.9 % sequence similarity) and the next closest recognized species was Desulfofrigus oceanense (93.5 %). On the basis of the significant differences in the 16S rRNA gene sequence and phenotypic characteristics between strain MSL71T and each of the related species, a novel genus and species, Desulfoluna butyratoxydans gen. nov., sp. nov., are proposed to accommodate strain MSL71T. The type strain is MSL71T (=JCM 14721T=DSM 19427T). PMID- 18398178 TI - Desulfotomaculum alcoholivorax sp. nov., a moderately thermophilic, spore forming, sulfate-reducer isolated from a fluidized-bed reactor treating acidic metal- and sulfate-containing wastewater. AB - A moderately thermophilic, Gram-positive, endospore-forming, sulfate-reducing bacterium was isolated from a fluidized-bed reactor treating acidic water containing metal and sulfate. The strain, designated RE35E1T, was rod-shaped and motile. The temperature range for growth was 33-51 degrees C (optimum 44-46 degrees C) and the pH range was 6.0-7.5 (optimum pH 6.4-7.3). The strain grew optimally without additional NaCl. The electron acceptors were 10 mM sulfate, thiosulfate and elemental sulfur and 1 mM (but not 10 mM) sulfite. Various alcohols and carboxylic acids were utilized as electron donors. Fermentative growth occurred on pyruvate. The cell wall contained meso-diaminopimelic acid, and the major respiratory isoprenoid quinone was menaquinone MK-7. The major whole-cell fatty acids were iso-C15 : 0, iso-C17 : 1 omega 10c and iso-C17 : 0. Strain RE35E1T was related to representatives of the genera Desulfotomaculum and Sporotomaculum, the closest relatives being Desulfotomaculum arcticum DSM 17038T (96.3 % 16S rRNA gene sequence similarity) and Sporotomaculum hydroxybenzoicum DSM 5475T (92.0 % similarity). Strain RE35E1T represents a novel species, for which the name Desulfotomaculum alcoholivorax sp. nov. is proposed. The type strain is RE35E1T (=DSM 16058T=JCM 14019T). PMID- 18398179 TI - Altererythrobacter indicus sp. nov., isolated from wild rice (Porteresia coarctata Tateoka). AB - A Gram-negative, rod-shaped, non-spore-forming organism, strain MSSRF26T, was isolated from mangrove-associated wild rice in India. On the basis of 16S rRNA gene sequence similarities, strain MSSRF26T was shown to belong to the Alphaproteobacteria, most closely related to Altererythrobacter luteolus and Altererythrobacter epoxidivorans (96.1 and 95.9 % similarity to the respective type strains). Chemotaxonomic data [major ubiquinones Q-10 (91 %) and Q-9 (9 %); major polyamine spermidine, with putrescine, cadaverine and spermine detected only in trace amounts; major polar lipids phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and sphingoglycolipid; major fatty acid C18 : 1 omega 7c and C(14 : 0) 2-OH as hydroxylated fatty acid] supported the affiliation of MSSRF26T to the genus Altererythrobacter. Fatty acid data and physiological and biochemical tests allowed phenotypic differentiation of the isolate from described Altererythrobacter species. Strain MSSRF26T therefore represents a novel species, for which the name Altererythrobacter indicus sp. nov. is proposed, with the type strain MSSRF26T (=LMG 23789T =DSM 18604T). PMID- 18398180 TI - Bacillus aurantiacus sp. nov., an alkaliphilic and moderately halophilic bacterium isolated from Hungarian soda lakes. AB - Three alkaliphilic and moderately halophilic strains designated K1-5T, K1-10 and B1-1, characterized by optimal growth at pH 9.0-10.0 and at 3-7 % (w/v) NaCl, were isolated from extremely shallow, alkaline soda lakes located in Hungary. Cells of the strains are Gram-positive, straight rods and form a central to subterminal, ellipsoidal endospore. The isolates are strictly aerobic, catalase positive, oxidase-negative and contain a peptidoglycan of type A1 gamma based on meso-diaminopimelic acid. In strain K1-5T, menaquinone-7 (MK-7) is the predominant isoprenoid quinone and anteiso-C15 : 0 is the major cellular fatty acid. The DNA G+C content of strain K1-5T is 42.9 mol%. 16S rRNA gene-based phylogenetic analysis revealed that the strains exhibit levels of sequence similarity of less than 95.8 % to known Bacillus species. According to the polyphasic characterization, the strains represent a novel species, for which the name Bacillus aurantiacus sp. nov. is proposed. The type strain is K1-5T (=DSM 18675T =CCM 7447T =NCAIM B002265T). PMID- 18398181 TI - Clostridium tepidiprofundi sp. nov., a moderately thermophilic bacterium from a deep-sea hydrothermal vent. AB - A moderately thermophilic, anaerobic bacterium (strain SG 508T) was isolated from a hydrothermal vent chimney located at 1 degrees N on the East Pacific Rise at a depth of 2650 m. Cells of strain SG 508T were straight to slightly curved rods, 0.4-0.6 microm in diameter and 2.0-3.0 microm in length. Spore formation was observed only below pH 5.5. The temperature range for growth was 22-60 degrees C, with optimum growth at 50 degrees C. The pH range for growth was 4.0-8.5, with optimum growth at pH 6.0-6.8. Growth of strain SG 508T was observed at NaCl concentrations ranging from 1.0 to 6.0 % (w/v), with optimum growth at 2.5 % (w/v). Substrates utilized by strain SG 508T included casein, peptone, tryptone, yeast extract, beef extract, starch, maltose and glucose. The products of glucose fermentation were ethanol, acetate, H2, formate and CO2. Strain SG 508T was able to reduce elemental sulfur to hydrogen sulfide. The DNA G+C content of strain SG 508T was 30.9 mol%. 16S rRNA gene sequence analysis revealed that the isolated organism belonged to cluster I of the genus Clostridium. On the basis of its physiological properties and data from phylogenetic analyses, strain SG 508T is considered to represent a novel species of the genus Clostridium, for which the name Clostridium tepidiprofundi sp. nov. is proposed. The type strain is SG 508T (=DSM 19306T =VKM B-2459T). PMID- 18398182 TI - Halosarcina pallida gen. nov., sp. nov., a halophilic archaeon from a low-salt, sulfide-rich spring. AB - A novel halophilic archaeon, strain BZ256T, was isolated from Zodletone Spring, a sulfide- and sulfur-rich spring in south-western Oklahoma, USA. Cells were non motile, non-flagellated cocci that divided along two axes, resulting in the formation of sarcina-like clusters. Strain BZ256T grew at salt concentrations ranging from 1.3 to 4.3 M NaCl, with optimum growth at approximately 3.4 M, and required at least 1 mM Mg2+ for growth. The pH range for growth was 5.0 to at least 8.5, and the temperature range for growth was 25-45 degrees C. The two diether phospholipids that are typical of members of the order Halobacteriales, namely phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, were present in strain BZ256T, as were two glycolipids chromatographically identical to S-DGD-1 and DGD-1. The 16S rRNA gene sequence of strain BZ256T showed 96.8 % similarity to that of the type strain of Halogeometricum borinquense, the closest recognized species within the order Halobacteriales. The DNA G+C content of strain BZ256T was 65.4 mol%. Microscopic, physiological, biochemical and phylogenetic comparisons between strain BZ256T and recognized genera of extremely halophilic archaea suggest that this strain represents a member of a novel genus and species within the family Halobacteriaceae, for which the name Halosarcina pallida gen. nov., sp. nov. is proposed. The type strain of Halosarcina pallida is BZ256T (=KCTC 4017T =JCM 14848T). PMID- 18398183 TI - Devosia chinhatensis sp. nov., isolated from a hexachlorocyclohexane (HCH) dump site in India. AB - A Gram-negative, motile, rod-shaped and non-spore-forming bacterium was isolated from a soil sample collected from the area adjoining an India Pesticide Limited plant, Lucknow, India. Strain IPL18T was characterized on the basis of phenotypic and genotypic data. Based on 16S rRNA gene sequence analysis, this strain was shown to belong to genus Devosia, with highest sequence similarity of 97.5 % to Devosia riboflavina DSM 7230T. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequence analysis revealed that strain IPL18T shows an evolutionary relationship with Devosia neptuniae J1T. Strain IPL18T contains C18 : 1 omega 7c, 11-methyl C18 : 1 omega 7c and C16 : 0 as the major fatty acids along with 3-OH C18 : 0, characteristic of the genus Devosia. The branched-chain fatty acid iso-H C18 : 1 (4.69 %), not present in any of the Devosia species known so far, has been reported in strain IPL18T. The results of DNA-DNA hybridization experiments (13 % relatedness with D. riboflavina DSM 7230T and 11 % with D. neptuniae J1T), phylogenetic analysis and biochemical tests confirm that strain IPL18T represents a novel species of the genus Devosia for which the name Devosia chinhatensis sp. nov. is proposed. The type strain is IPL18T (=CCM 7426T=MTCC 8593T). PMID- 18398184 TI - Neptunomonas japonica sp. nov., an Osedax japonicus symbiont-like bacterium isolated from sediment adjacent to sperm whale carcasses off Kagoshima, Japan. AB - Novel bacterial species were isolated from sediments adjacent to sperm whale carcasses off Kagoshima, Japan, at a depth of 226-246 m. The isolated strains, JAMM 0745T, JAMM 1380, JAMM 1475 and JAMM 1610, were Gram-negative, rod-shaped, non-spore-forming and motile by means of a single polar or subterminal flagellum. Phylogenetic analysis based on 16S rRNA gene sequences of the novel isolates indicated a relationship to a symbiotic bacterial clone of the polychaete Osedax japonicus (99.6-99.9 % sequence similarity) and these bacteria were closely related to members of the genus Neptunomonas (95.6-96.0 % similarity) within the class Gammaproteobacteria. The novel strains were able to produce isoprenoid quinone Q-8 as the major quinone component. The predominant fatty acids were C16 : 0, C16 : 1 and C18 : 1, with C18 : 2 and C20 : 2 present in smaller amounts. The DNA G+C contents of the four novel strains were about 43.6-43.8 mol%. Based on the taxonomic differences observed, the four isolated strains appear to represent a novel species of the genus Neptunomonas. The name Neptunomonas japonica sp. nov. (type strain JAMM 0745T=JCM 14595T=DSM 18939T) is proposed for the novel strains. PMID- 18398185 TI - Halomonas nitroreducens sp. nov., a novel nitrate- and nitrite-reducing species. AB - We have carried out a polyphasic taxonomic study of strain 11ST, a halophilic, Gram-negative bacterium that is able to respire on nitrate and nitrite in anaerobiosis. Strain 11ST was isolated from a solar saltern in Cahuil, a region next to Pichilemu (Chile). It grows at NaCl concentrations within the range of 3 20 % w/v (optimum 5-7.5 %), temperatures from 4 to 45 degrees C (optimum 20-32 degrees C) and within a pH range of 5-10 (optimum pH 7-9). Its 16S rRNA gene sequence indicates that it belongs to the genus Halomonas in the class Gammaproteobacteria. Its closest relatives are Halomonas alimentaria, H. denitrificans, H. organivorans and H. ventosae, with the type strains of which our strain showed maximum 16S rRNA gene sequence similarity values of 97.1-98.1 %. Its G+C content is 65.3 mol%. DNA-DNA hybridization studies showed 54.2 % relatedness between strain 11ST and H. alimentaria DSM 15356T and 47.2 % relatedness between strain 11ST and H. organivorans CECT 5995T. Lower DNA-DNA hybridization percentages were obtained against the type strains of other related Halomonas species. Its major fatty acids are C12 : 0 3-OH (5.56 %), iso-C15 : 0 2 OH/C16 : 1 omega 7c (22.30 %), C16 : 0 (27.80 %) and C18 : 1 omega 7c (29.92 %). The proposed name for the novel species is Halomonas nitroreducens sp. nov., with strain 11ST (=CECT 7281T =LMG 24185T) being the type strain. PMID- 18398186 TI - Chromobacterium aquaticum sp. nov., isolated from spring water samples. AB - Strain CC-SEYA-1T, a motile, Gram-negative, non-violet-pigmented bacterium, was isolated on nutrient agar from spring-water samples collected from Yang-Ming Mountain, Taipei County, Taiwan. 16S rRNA gene sequence studies showed that the strain clustered with Chromobacterium violaceum (96.8 % similarity) and Chromobacterium subtsugae (96.5 % similarity), followed by Aquitalea magnusonii (95.8 % similarity). The fatty acid profile was slightly different from those reported for C. violaceum, C. subtsugae and A. magnusonii. The results of DNA-DNA hybridization, and physiological and biochemical tests allowed both genotypic and phenotypic differentiation of the isolate from the described Chromobacterium species. It is evident from the data obtained that the strain should be classified as a novel species in the genus Chromobacterium. The name proposed for this taxon is Chromobacterium aquaticum sp. nov.; the type strain is CC-SEYA-1T (=CCUG 55175T=BCRC 17769T). PMID- 18398187 TI - Tenacibaculum soleae sp. nov., isolated from diseased sole (Solea senegalensis Kaup). AB - A novel Gram-negative, rod-shaped, gliding bacterial strain designated LL04 12.1.7T was isolated from diseased sole (Solea senegalensis Kaup) in Galicia, Spain. Colonies were yellow-pigmented with uneven edges and did not adhere to the agar. The DNA G+C content of the strain was 29.8 mol%. 16S rRNA gene sequence similarity analysis indicated that strain LL04 12.1.7T is a member of the genus Tenacibaculum in the family Flavobacteriaceae. Sequence similarities between the isolate and the type strains of other members of the genus were 96.7-94.8 %. The major fatty acids (>10 % of total fatty acids) were iso-C15 : 0 (23.1 %), iso-C15 : 0 3-OH (10.6 %), C15 : 1 omega 6c (12.2 %) and summed feature 3 (comprising C16 : 1 omega 7c and/or iso-C15 : 0 2-OH, 11.0 %). Genotypic and phenotypic data distinguished strain LL04 12.1.7T from the 11 recognized Tenacibaculum species, indicating that it represents a novel species, for which the name Tenacibaculum soleae sp. nov. is proposed. The type strain is strain LL04 12.1.7T (=CECT 7292T =NCIMB 14368T). PMID- 18398188 TI - Halobacillus alkaliphilus sp. nov., a halophilic bacterium isolated from a salt lake in Fuente de Piedra, southern Spain. AB - A Gram-positive, spore-forming, halophilic bacterial strain, FP5T, was isolated from a salt lake in southern Spain and subjected to a polyphasic taxonomic study. Strain FP5T was strictly aerobic. Cells were coccoidal, occurring singly or in clusters. The cell-wall peptidoglycan type of strain FP5T was A4 beta based on l Orn-d-Asp. Strain FP5T was characterized chemotaxonomically by having MK-7 as the major menaquinone and anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C16 : 0 as the main fatty acids. The isolate grew optimally at 37 degrees C and in presence of 10 % NaCl; no growth was observed in the absence of NaCl. The DNA G+C content was 43.5 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain FP5T falls within the evolutionary radiation of species of the genus Halobacillus. Levels of 16S rRNA gene sequence similarity between strain FP5T and the type strains of nine recognized Halobacillus species were in the range 97.0-99.0 %. Levels of DNA-DNA relatedness indicated that strain FP5T represents a genomic species that is distinct from recognized Halobacillus species. Strain FP5T could be differentiated from recognized Halobacillus species based on several phenotypic characteristics. On the basis of phenotypic, phylogenetic and genomic data, strain FP5T is considered to represent a novel species of the genus Halobacillus, for which the name Halobacillus alkaliphilus sp. nov. is proposed. The type strain is FP5T (=DSM 18525T =ATCC BAA-1361T). PMID- 18398189 TI - Lactobacillus ceti sp. nov., isolated from beaked whales (Ziphius cavirostris). AB - Biochemical and molecular genetic studies were performed on three isolates of an unknown Gram-positive, catalase-negative and rod-shaped organism isolated from the lungs and liver of two beaked whales. The organisms were tentatively identified as Lactobacillus spp. based on cellular morphology and biochemical tests. 16S rRNA gene sequencing studies confirmed the provisional identification of the novel isolates as members of the genus Lactobacillus, but the isolates did not correspond to any recognized species of this genus. The novel strains shared the same phenotypic characteristics and exhibited 100 % 16S rRNA gene sequence similarity. The nearest phylogenetic relatives of the novel isolates were Lactobacillus satsumensis DSM 16230T (94.2 % 16S rRNA gene sequence similarity), Lactobacillus salivarius JCM 1047 (94.0 %), Lactobacillus nagelii ATCC 700692T (94.0 %) and Lactobacillus saerimneri DSM 16049T (93.8 %). The novel isolates could be distinguished from these species and other related species of the genus Lactobacillus by physiological and biochemical tests. On the basis of these phenotypic, physiological and phylogenetic findings, it is proposed that the new isolates from whales be classified as a novel species of the genus Lactobacillus, Lactobacillus ceti sp. nov. The type strain is 142-2T (=CECT 7185T=CCUG 53626T). PMID- 18398190 TI - Simiduia agarivorans gen. nov., sp. nov., a marine, agarolytic bacterium isolated from shallow coastal water from Keelung, Taiwan. AB - A Gram-negative, heterotrophic, agarolytic, marine bacterium, designated strain SA1T, was isolated from a seawater sample collected in the shallow coastal region of Keelung, Taiwan. Cells were straight to slightly curved rods. Nearly all of the cells were non-motile and non-flagellated during the exponential phase of growth in broth cultures; a few cells (<1 %) were motile and were considered to have monotrichous flagella. The isolate required NaCl for growth and grew optimally at 30-35 degrees C and 2-3 % (w/v) NaCl. It grew aerobically and was incapable of anaerobic growth by fermentation of glucose or other carbohydrates. However, anaerobic growth could be achieved by reduction of nitrate to nitrite. Polar lipids comprised phosphatidylethanolamine (71.8 %), diphosphatidylglycerol (12.7 %), phosphatidylglycerol (12.2 %) and phosphatidylserine (3.3 %). Isoprenoid quinones consisted of Q-10 (87.5 %), MK-9 (6.6 %) and MK-7 (5.9 %). Major cellular fatty acids were C16 : 1 omega 7c and/or iso-C15 : 0 2-OH (28.6 %), C17 : 1 omega 8c (22.8 %), C16 : 0 (14.5 %), C18 : 1 omega 7c (11.0 %) and C17 : 0 (6.4 %). The DNA G+C content was 55.6 mol%. Phylogeny based on 16S rRNA gene sequence analysis showed that strain SA1T formed a distinct lineage within the class Gammaproteobacteria. Strain SA1T was related most closely to Teredinibacter turnerae, Cellvibrio spp., Saccharophagus degradans, Pseudomonas spp. and Microbulbifer spp., strains of these species sharing <93 % 16S rRNA gene sequence similarity with strain SA1T. The phylogenetic data and those from physiological, morphological and chemotaxonomic characterizations indicated that strain SA1T represents a novel species and genus, for which the name Simiduia agarivorans gen. nov., sp. nov. is proposed. The type strain is SA1T (=BCRC 17597T=JCM 13881T). PMID- 18398191 TI - Alkalibacterium indicireducens sp. nov., an obligate alkaliphile that reduces indigo dye. AB - Indigo-reducing, obligately alkaliphilic strains A11T, F11 and F12 were isolated from indigo fermentation liquor obtained from Tokushima Prefecture, Shikoku, Japan. The isolates grew at pH 9.0-12.3, but not at pH 7.0-8.0. The optimum pH range for growth was 9.5-11.5. They were Gram-negative, facultatively anaerobic, rod-shaped strains with peritrichous flagella. The isolates grew in 0-14 % (w/v) NaCl, with optimum growth at 1-11 %. They grew at temperatures of 15-35 degrees C with optimum growth at around 20-30 degrees C. dl-Lactate was the major end product from d-glucose. No quinones were detected. The peptidoglycan type was A4 alpha, l-Lys (l-Orn)-d-Asp. The major cellular fatty acids were C16 : 0, C16 : 17c and C18 : 19c. The DNA G+C contents were 47.0-47.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequence data indicated that the isolates belong to the genus Alkalibacterium. DNA-DNA hybridization revealed low relatedness values between the isolates and the three phylogenetically most closely related species, Alkalibacterium olivapovliticus, Alkalibacterium psychrotolerans and Alkalibacterium iburiense (<41 %). On the basis of phenotypic characteristics, including hydrolysis of cellulose and fermentation of carbohydrates, and chemotaxonomic characteristics, phylogenetic data and DNA-DNA relatedness data, it is concluded that the isolates merit classification as representatives of a novel species of the genus Alkalibacterium, for which the name Alkalibacterium indicireducens sp. nov. is proposed. The type strain of this species is A11T (=JCM 14232T=NCIMB 14253T). PMID- 18398192 TI - Saxeibacter lacteus gen. nov., sp. nov., an actinobacterium isolated from rock. AB - A Gram-positive, cream-coloured, non-motile, rod-shaped actinomycete, designated DLS-10T, was isolated from a rock collected on the peak of Darangshi Oreum (a small mountain 300 m above sea level) in Jeju, Republic of Korea. The temperature and pH ranges for growth were 4-37 degrees C and pH 5.1-9.1. The organism grew at NaCl concentrations up to 3 % and had the following chemotaxonomic characteristics: meso-diaminopimelic acid was the diagnostic diamino acid in the cell-wall peptidoglycan, the polar lipids included diphosphatidylglycerol, phosphatidylmethylethanolamine and two unknown phospholipids, the major menaquinone was MK-8(H4) and the major fatty acids were anteiso-C15 : 0, C16 : 0, iso-C16 : 0 and anteiso-C17 : 0. The DNA G+C content was 74.3 mol%. Phylogenetic analyses based on 16S rRNA gene sequences revealed that the organism was related to members of the family Nakamurellaceae in the suborder Frankineae. The most closely related described organisms were the type strains of Humicoccus flavidus (97.1 % sequence similarity) and Nakamurella multipartita (95.2 %). On the basis of the morphological, cultural, physiological, chemotaxonomic and phylogenetic evidence presented here, the organism represents a novel genus and species of the family Nakamurellaceae, for which the name Saxeibacter lacteus gen. nov., sp. nov. is proposed. The type strain of Saxeibacter lacteus is DLS-10T (=KCTC 19285T =DSM 19367T). PMID- 18398193 TI - Lactobacillus sobrius Konstantinov et al. 2006 is a later synonym of Lactobacillus amylovorus Nakamura 1981. AB - While studying the taxonomy of six lactic acid bacterium isolates from Finnish porcine intestine and faeces, the taxonomic positions of Lactobacillus sobrius type strain DSM 16698T and strain AD5 based on comparative 16S rRNA sequence analysis were found to be controversial, as they showed high similarity to Lactobacillus amylovorus strains. Therefore, the taxonomy of these species was addressed in a polyphasic taxonomy study that included, in addition to re evaluating the 16S rRNA gene sequence and DNA-DNA reassociation results, multilocus sequence analysis (MLSA) of the housekeeping genes encoding the phenylalanyl-tRNA synthase alpha subunit (pheS) and RNA polymerase alpha subunit (rpoA) as well as numerical analysis of HindIII and EcoRI ribotypes. 16S rRNA gene sequence analysis demonstrated a very high similarity between the L. sobrius and L. amylovorus type and reference strains and representative Finnish porcine isolates (99.6-99.9 %). The MLSA data showed the close phylogenetic relationship of these strains; pheS and rpoA gene sequence similarities were 98.5-100 % and 99.6-99.8 %, respectively. Numerical analyses of HindIII/EcoRI ribotypes placed these strains in a single cluster by both enzymes. Finally, the DNA-DNA reassociation experiments revealed high reassociation levels (higher than 79 %) between the strains. These results indicate that DSM 16698T, AD5 and the related porcine lactobacilli strains from Finland constitute a single species, Lactobacillus amylovorus, and that the name Lactobacillus sobrius should be considered as a later synonym of Lactobacillus amylovorus. PMID- 18398194 TI - Lactobacillus equigenerosi sp. nov., a coccoid species isolated from faeces of thoroughbred racehorses. AB - Two strains of lactic acid bacteria were isolated from faeces of two actively racing thoroughbred horses. The isolates formed a subcluster in the Lactobacillus reuteri phylogenetic group, closely related to Lactobacillus fermentum, L. gastricus, L. ingluviei and L. mucosae, by phylogenetic analysis based on 16S rRNA gene sequences. Levels of DNA-DNA relatedness revealed that the isolates belonged to the same taxon and were genetically separated from their phylogenetic relatives. Biochemical and physiological characteristics also distinguished the isolates from their phylogenetic relatives. The isolates produced spherical or oval cells, and tetrad-like cells were rarely seen. To the best of our knowledge, this is the first report of this morphological characteristic within the genus Lactobacillus. Thus, the isolates represent an atypical novel species of the genus Lactobacillus, for which the name Lactobacillus equigenerosi sp. nov. is proposed. The type strain is NRIC 0697T (=JCM 14505T =DSM 18793T). PMID- 18398195 TI - Bacillus coahuilensis sp. nov., a moderately halophilic species from a desiccation lagoon in the Cuatro Cienegas Valley in Coahuila, Mexico. AB - A moderately halophilic, Gram-positive and rod-shaped bacterium, strain m4-4T, was isolated from a Chihuahuan desert lagoon in Cuatro Cienegas, Coahuila, Mexico. Strain m4-4T was found to grow optimally at 30-37 degrees C, pH 7.0-8.0 and 5 % NaCl and to tolerate from 0.5 % to 10 % NaCl. It was shown to be aerobic. The genomic DNA G+C content was about 37 mol%. Strain m4-4T exhibited minimal or no growth on most sugars tested. Its major cellular fatty acids were C14 : 0, C16 : 0 and C18 : 1. Based on phylogenetic analysis of 16S rRNA and recA gene sequences, we observed that the closest relatives of the isolate are moderately halophilic Bacillus species, with 16S rRNA gene sequence similarity ranging from 96.6 to 97.4 % (Bacillus marisflavi, Bacillus aquimaris and Bacillus vietnamensis). Additionally, using genomic data it was determined that the type strain contains a total of nine rRNA operons with three slightly different sequences. On the basis of phenotypic and molecular properties, strain m4-4T represents a novel species within the genus Bacillus, for which the name Bacillus coahuilensis sp. nov. is proposed, with the type strain m4-4T (=NRRL B-41737T =CECT 7197T). PMID- 18398196 TI - Micromonospora chaiyaphumensis sp. nov., isolated from Thai soils. AB - Three actinomycete strains, MC5-1T, MC7-1 and R1-1, were isolated from soil samples collected in Thailand. Their taxonomic positions were determined using a polyphasic approach. The chemotaxonomic characteristics of these strains coincided with those of the genus Micromonospora, i.e. meso-diaminopimelic acid and N-glycolyl muramic acid were present in the cell-wall peptidoglycan, the whole-cell sugars were of pattern D, the phospholipids were of type II and the cellular fatty acids were of type 3b. Phylogenetic analysis of the 16S rRNA gene sequences revealed a close relationship between strains MC5-1T, MC7-1 and R1-1 (99.8 % sequence similarity) and Micromonospora auratinigra JCM 12357T (99.3 %). The three novel strains were clearly distinguishable from M. auratinigra JCM 12357T from the low DNA-DNA relatedness (< or =43.4 %). On the basis of the data presented, strain MC5-1T represents a novel species of the genus Micromonospora, for which the name Micromonospora chaiyaphumensis is proposed. The type strain is MC5-1T (=KCTC 19332T=JCM 12873T=PCU 267T=TISTR 1564T). PMID- 18398197 TI - Methanocella paludicola gen. nov., sp. nov., a methane-producing archaeon, the first isolate of the lineage 'Rice Cluster I', and proposal of the new archaeal order Methanocellales ord. nov. AB - A novel mesophilic, hydrogenotrophic methanogen, strain SANAET, was isolated from an anaerobic, propionate-degrading enrichment culture, which was originally established from rice paddy soil. The cells were non-motile, Gram-negative and rod-shaped (1.8-2.4 microm long by 0.3-0.6 microm wide). Growth of strain SANAET was observed at 25-40 degrees C, with an optimum temperature range for growth of 35-37 degrees C. The pH range for growth was 6.5-7.8, with an optimum at pH 7.0. The salinity range for growth was 0-1 g NaCl l(-1) (0-17 mM). The isolate was able to utilize H2/CO2 and formate for growth and methane production. The G+C content of the genomic DNA was 56.6 mol%. Based on comparative 16S rRNA gene sequence analysis, strain SANAET was affiliated with a clone lineage of the Archaea, Rice Cluster I (RC-I), placing it between the orders Methanosarcinales and Methanomicrobiales within the class 'Methanomicrobia'. 16S rRNA gene sequence similarities between strain SANAET and members of Methanosarcinales were in the range 80.0-82.8 %, and those between the strain and members of Methanomicrobiales ranged from 77.5 to 82.4 %. In addition to 16S rRNA gene analysis, sequence analysis of the mcrA gene (encoding the alpha subunit of methyl-coenzyme M reductase, a key enzyme in the methane production pathway) also showed that strain SANAET was affiliated with the RC-I lineage. Here, we propose the name Methanocella paludicola gen. nov., sp. nov. for the isolate, the first of the RC I lineage. The type strain is SANAET (=JCM 13418T=NBRC 101707T=DSM 17711T). In addition, we also propose the status of order for the RC-I lineage, for which we propose the name Methanocellales ord. nov. PMID- 18398198 TI - Reclassification of Acinetobacter grimontii Carr et al. 2003 as a later synonym of Acinetobacter junii Bouvet and Grimont 1986. AB - Using tDNA-PCR, the type strain CCM 7198T (<--CIP 107470T <--17A04T) of Acinetobacter grimontii was found to be indistinguishable from Acinetobacter junii strains. Therefore, the phenotypic properties, amplified fragment length polymorphism (AFLP) patterns and 16S rRNA and rpoB gene sequences of the type strain of A. grimontii (CCM 7198T) were determined. We found that the strain used l-arginine and l-glutamate, in contrast to the original description and in accordance with the phenotypic properties of A. junii. By AFLP analysis, A. grimontii CCM 7198T clustered at 50.2 % with a set of A. junii strains previously identified by DNA-DNA hybridization, which is in accordance with the previously established intraspecies values of this technique. Sequence similarity of the 16S rRNA gene between the type strains of the two species was found to be 99.9 %. Finally, DNA-DNA relatedness between the type strains of A. junii and A. grimontii was redetermined and was found to be 85 %. These findings were corroborated for a second representative of the A. grimontii type strain, DSM 14968T. These data confirm that Acinetobacter grimontii is a later heterotypic synonym of Acinetobacter junii. PMID- 18398199 TI - Hymenobacter soli sp. nov., isolated from grass soil. AB - A Gram-negative, strictly aerobic, non-spore-forming, rod-shaped, red-pink bacterium, designated strain PB17T, was isolated from grass soil in the Republic of Korea and its taxonomic position was investigated by means of a polyphasic approach. Phylogenetic analysis of 16S rRNA gene sequences showed that strain PB17T belonged to the genus Hymenobacter. Sequence similarities between strain PB17T and the type strains of Hymenobacter species with validly published names ranged from 90.1 to 91.8 %. The predominant cellular fatty acids in strain PB17T were iso-C15 : 0, summed feature 4 (iso-C15 : 0 2-OH and/or C16 : 1 omega 7c), C16 : 1 omega 5c and anteiso-C15 : 0. The DNA G+C content was 58.8 mol%. The results of phylogenetic, chemotaxonomic and phenotypic characterization indicated that strain PB17T can be distinguished from all known Hymenobacter species and represents a novel species, for which the name Hymenobacter soli sp. nov. is proposed. The type strain is PB17T (=KCTC 12607T =LMG 24240T). PMID- 18398200 TI - Azonexus hydrophilus sp. nov., a nifH gene-harbouring bacterium isolated from freshwater. AB - Three Gram-negative, non-pigmented, rod-shaped, facultatively aerobic bacterial strains, designated d8-1T, d8-2 and IMCC1716, were isolated from a freshwater spring sample and a eutrophic freshwater pond. Based on characterization using a polyphasic approach, the three strains showed highly similar phenotypic, physiological and genetic characteristics. All of the strains harboured the nitrogenase gene nifH, but nitrogen-fixing activities could not be detected in nitrogen-free culture media. The three strains shared 99.6-99.7 % 16S rRNA gene sequence similarity and showed 89-100 % DNA-DNA relatedness, suggesting that they represent a single genomic species. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains d8-1T, d8-2 and IMCC1716 formed a monophyletic branch in the periphery of the evolutionary radiation occupied by the genus Azonexus. Their closest neighbours were Azonexus caeni Slu-05T (96.7-96.8 % similarity) and Azonexus fungiphilus BS5-8T (96.3-96.6 %). The DNA-DNA relatedness of the novel strains to these two species of the genus Azonexus was less than 70 %. The isolates could also be differentiated from recognized members of the genus Azonexus on the basis of phenotypic and biochemical characteristics. It is evident, therefore, that the three strains represent a novel species of the genus Azonexus, for which the name Azonexus hydrophilus sp. nov. is proposed. The type strain is d8-1T (=LMG 24005T=BCRC 17657T). PMID- 18398201 TI - Yersinia similis sp. nov. AB - Strains originally identified phenotypically as members of the species Yersinia pseudotuberculosis were subjected to a more detailed classification employing 16S rRNA gene sequence analysis, DNA-DNA hybridization, determination of the DNA base composition and phenotypic characterization. The quinone system, consisting of the predominant compound ubiquinone Q-8 and minor amounts of menaquinone MK-8, the major components of the polar lipid profile, as well as the polyamine pattern, with putrescine as the major compound, supported the assignment of the strains to the genus Yersinia. Based on DNA-DNA relatedness, a specific 16S rRNA gene sequence type, absence of melibiose fermentation and a polar lipid profile lacking phosphatidylmonomethylethanolamine and two aminolipids, the strains were identified as members of a novel species for which the name Yersinia similis sp. nov. is proposed. The type strain of Yersinia similis sp. nov. is strain Y228T (=CCUG 52882T=LMG 23763T). PMID- 18398202 TI - Azospirillum rugosum sp. nov., isolated from oil-contaminated soil. AB - The taxonomic status of a light-orange-coloured bacterial isolate from an oil contaminated soil sample was characterized by using a polyphasic taxonomic approach. Comparative analysis of 16S rRNA gene sequences demonstrated that the isolate belonged phylogenetically to the genus Azospirillum, with Azospirillum canadense, Azospirillum brasilense and Azospirillum doebereinerae as its closest phylogenetic relatives (97.3, 97.0 and 97.0 % similarity, respectively). DNA-DNA pairing studies showed that the unidentified organism displayed 25.0, 17.0 and 19.0 % relatedness to the type strains of A. brasilense, A. canadense and A. doebereinerae, respectively. The generic assignment was confirmed by chemotaxonomic data, which revealed a fatty acid profile that was characteristic of the genus Azospirillum, consisting of straight-chain saturated and unsaturated fatty acids with C18 : 1 omega 7c as the major fatty acid, and ubiquinone with ten isoprene units (Q-10) as the predominant respiratory quinone. On the basis of both the phenotypic and molecular genetic evidence, it is proposed that the unknown isolate be classified as a representative of a novel species of the genus Azospirillum, for which the name Azospirillum rugosum sp. nov. is proposed. The type strain is IMMIB AFH-6T (=CCUG 53966T=DSM 19657T). PMID- 18398203 TI - Lutispora thermophila gen. nov., sp. nov., a thermophilic, spore-forming bacterium isolated from a thermophilic methanogenic bioreactor digesting municipal solid wastes. AB - A novel anaerobic, moderately thermophilic, spore-forming, rod-shaped bacterium (strain EBR46T) was isolated from an enrichment culture derived from an anaerobic thermophilic (55 degrees C) methanogenic bioreactor treating artificial solid wastes. Phylogeny based on 16S rRNA gene sequence analysis placed strain EBR46T within a distinct lineage between Clostridium clusters II and III. The closest recognized relative of strain EBR46T was Gracilibacter thermotolerans DSM 17427T (85.3 % 16S rRNA gene sequence similarity). The DNA G+C content of strain EBR46T was 36.2 mol%. The novel strain grew optimally at 55-58 degrees C and at pH 7.5 8.0 and was able to grow on peptone, tryptone, Casamino acids, casein hydrolysate, methionine, threonine, tryptophan, cysteine, lysine and serine in the presence of 0.2 % yeast extract. Carbohydrates were not utilized. The main products from tryptone utilization were acetate, iso-butyrate, propionate and iso valerate. Strain EBR46T produced hydrogen sulfide from cysteine. The major fatty acids were iso-C15 : 0, C14 : 0, C16 : 0 DMA (dimethyl acetal) and iso-C15 : 0 DMA. Based on its unique phylogenetic and physiological features, strain EBR46T is considered to represent a novel species of a new genus, for which the name Lutispora thermophila gen. nov., sp. nov. is proposed. The type strain of the type species is EBR46T (=NBRC 102133T=DSM 19022T). PMID- 18398204 TI - Sutterella parvirubra sp. nov. and Megamonas funiformis sp. nov., isolated from human faeces. AB - Three strains of anaerobic, non-spore-forming, Gram-negative coccobacilli (YIT 11816T, YIT 11817 and YIT 11818) were isolated from human faeces. On the basis of 16S rRNA gene sequence similarity, these strains were shown to belong to the family Alcaligenaceae and to be related to the type strain of Sutterella stercoricanis (94.9 %) and to Sutterella wadsworthensis WAL 7877 (94.3 %); the similarity to strains of any other species with a validly published name within the family Alcaligenaceae was less than 92 %. Biochemical data supported the affiliation of these strains to the genus Sutterella. These strains therefore represent a novel species, for which the name Sutterella parvirubra sp. nov. is proposed; the type strain is YIT 11816T (=DSM 19354T =JCM 14724T). The cells of another isolate, strain YIT 11815T, were non-spore-forming, Gram-negative, very large rods, 1x5-200 microm in size, with or without a central, subterminal or terminal swelling of 2-4 microm diameter when grown in a broth medium supplemented with glucose. Based on comparative 16S rRNA gene sequencing, this bacterium is a member of the family Acidaminococcaceae, and most closely related to Megamonas hypermegale (95.3 % similarity to the type strain). Interestingly, the 16S rRNA gene sequence of strain YIT 11815T showed 99 % similarity to sequences of uncultured colonic bacteria. A 16S rRNA gene sequence divergence value of >3 % from known cultured species suggested that isolate YIT 11815T represents a novel species, for which the name Megamonas funiformis sp. nov. is proposed; the type strain is YIT 11815T (=DSM 19343T =JCM 14723T). PMID- 18398205 TI - Wohlfahrtiimonas chitiniclastica gen. nov., sp. nov., a new gammaproteobacterium isolated from Wohlfahrtia magnifica (Diptera: Sarcophagidae). AB - New Gammaproteobacteria were isolated from 3rd stage fly larvae of the parasitic fly Wohlfahrtia magnifica. Phylogenetic analysis of the new isolates showed that these bacteria belong to a distinct lineage close to Ignatzschineria larvae, which was originally isolated from the same species of fly. The low similarity values in 16S rRNA gene sequences (93.8-94.8 %), and differences in fatty acid profiles, RiboPrint patterns, MALDI-TOF mass spectra of cell extracts, and physiological and biochemical characteristics differentiate the isolates from the type strain of Ignatzschineria larvae (DSM 13226T), and indicate that our isolates represent a new genus within the Gammaproteobacteria. The major isoprenoid quinone of the strains is Q8, the major fatty acids are C18 : 1 and C14 : 0, and the predominant polar lipids are phosphatidylglycerol, phosphatidylethanolamine and phosphatidylserine. The G+C content of the DNA of the type strain is 44.3 mol%. The name Wohlfahrtiimonas chitiniclastica gen. nov., sp. nov., is proposed for this novel genus and species. The type strain is S5T (=DSM 18708T=CCM 7401T). PMID- 18398206 TI - Limibacter armeniacum gen. nov., sp. nov., a novel representative of the family 'Flammeovirgaceae' isolated from marine sediment. AB - Two strictly aerobic, Gram-negative, gliding, apricot- to pale-orange-pigmented, rod-shaped bacteria, strains YM11-185T and YM11-159, were isolated from marine sediments in the Republic of Palau. Preliminary analysis based on the 16S rRNA gene sequence revealed that the novel isolates could be affiliated with the family 'Flammeovirgaceae' of the phylum Bacteroidetes. Strains YM11-185T and YM11 159 shared 100 % sequence similarity and showed less than 90 % similarity to cultivated representatives of the family 'Flammeovirgaceae'. The novel isolates are phenotypically and physiologically different from strains described previously. The G+C content of DNA was 27.8-27.9 mol%, MK-7 was the major menaquinone and iso-C15 : 0 and C16 : 1 omega 7c were the major fatty acids. On the basis of polyphasic taxonomic studies, it was concluded that strains YM11 185T and YM11-159 represent a novel species in a new genus of the family 'Flammeovirgaceae', for which the name Limibacter armeniacum gen. nov., sp. nov. is proposed. The type strain of Limibacter armeniacum is YM11-185T (=MBIC08286T =KCTC 22132T). PMID- 18398207 TI - Aeromicrobium ponti sp. nov., isolated from seawater. AB - A novel actinomycete strain, designated HSW-1T, was isolated from seawater collected from Hwasun Beach on the coast of Jeju Island, Republic of Korea. The cells were aerobic, Gram-positive, oxidase-negative, catalase-positive, non motile rods. The colonies were circular, smooth, convex and yellow in colour. The cell-wall peptidoglycan of this organism contained ll-diaminopimelic acid as the diagnostic diamino acid. The polar lipids included phosphatidylinositol, diphosphatidylglycerol, phosphatidylglycerol and three unknown phospholipids. The predominant menaquinone was MK-9(H4). The major fatty acids were C18 : 1 omega 9c, C16 : 0, C16 : 0 2-OH and 10-methyl C18 : 0. The DNA G+C content was 74.0 mol%. A neighbour-joining tree based on 16S rRNA gene sequences showed that this organism falls within the radiation of the genus Aeromicrobium. The closest phylogenetic neighbours were the type strains of Aeromicrobium erythreum (98.2 % 16S rRNA gene sequence similarity), Aeromicrobium alkaliterrae (97.9 %), Aeromicrobium marinum (97.3 %) and Aeromicrobium fastidiosum (97.0 %). The DNA DNA relatedness values between the novel isolate and its closest relative, A. erythreum DSM 8599T, ranged between 32 and 36 %. On the basis of the phenotypic and DNA-DNA hybridization data, strain HSW-1T represents a novel species of the genus Aeromicrobium, for which the name Aeromicrobium ponti sp. nov. is proposed. The type strain is HSW-1T (=DSM 19178T=KACC 20565T). PMID- 18398208 TI - Rubritalea sabuli sp. nov., a carotenoid- and squalene-producing member of the family Verrucomicrobiaceae, isolated from marine sediment. AB - The taxonomic status of a verrucomicrobial strain isolated from marine sediment was established based on a polyphasic examination. The novel isolate, strain YM29 052T, was obligately aerobic, Gram-negative, non-motile, coccoid or rod-shaped and chemoheterotrophic. Phylogenetic analyses based on 16S rRNA gene sequences demonstrated that the new isolate shared approximately 94-99 % sequence similarity with members of genus Rubritalea of the family Verrucomicrobiaceae within the phylum 'Verrucomicrobia'. Genomic DNA-DNA hybridization between strain YM29-052T and Rubritalea squalenifaciens HOact23T showed relatedness of <70 %, the value commonly accepted as the threshold for the phylogenetic definition of a species. Strain YM29-052T produces carotenoid compounds that render the cell biomass a pink colour; the strain also contains squalene. The cell-wall peptidoglycan of the novel strain contains muramic acid and meso-diaminopimelic acid. The DNA G+C content of strain YM29-052T was 47.7 mol%; MK-8 and MK-9 were the major menaquinones. The presence of iso-C14 : 0, iso-C16 : 0 and C16 : 1 omega 7c as major cellular fatty acids supported the identification of the novel isolate as a member of the genus Rubritalea. On the basis of polyphasic taxonomic evidence, it was concluded that strain YM29-052T should be classified within a novel species of the genus Rubritalea, for which the name Rubritalea sabuli sp. nov. is proposed. The type strain is YM29-052T (=MBIC08323T =KCTC 22127T). PMID- 18398209 TI - Description of Persicirhabdus sediminis gen. nov., sp. nov., Roseibacillus ishigakijimensis gen. nov., sp. nov., Roseibacillus ponti sp. nov., Roseibacillus persicicus sp. nov., Luteolibacter pohnpeiensis gen. nov., sp. nov. and Luteolibacter algae sp. nov., six marine members of the phylum 'Verrucomicrobia', and emended descriptions of the class Verrucomicrobiae, the order Verrucomicrobiales and the family Verrucomicrobiaceae. AB - Ten pale-pink- and pale-yellow-pigmented, Gram-negative, non-motile, rod-shaped, chemoheterotrophic bacteria designated strains YM20-087T, YM21-151, MN1-741T, YM27-120T, YM26-010T, YM24-184, YM20-122, A4T-83T, A5J-41-2T and A5J-40 were isolated from various marine environments and were subjected to a polyphasic taxonomic investigation. Phylogenetic analyses based on 16S rRNA gene sequences indicated that these isolates belonged to the phylum 'Verrucomicrobia' (subdivision 1) and represented three independent lineages that were distinct from species of genera of the family Verrucomicrobiaceae with validly published names. The cell-wall peptidoglycan of these strains contained muramic acid and meso-diaminopimelic acid. Strains MN1-741T, YM27-120T, YM26-010T, YM24-184 and YM20-122 produced pinkish carotenoid pigments. On the basis of polyphasic taxonomic evidence, it was concluded that these strains should be classified within three new genera, Persicirhabdus gen. nov. (with one species, the type species Persicirhabdus sediminis sp. nov.), Roseibacillus gen. nov. (with three species; type species Roseibacillus ishigakijimensis sp. nov.) and Luteolibacter gen. nov. (with two species; type species Luteolibacter pohnpeiensis sp. nov.), of the family Verrucomicrobiaceae within the phylum 'Verrucomicrobia'. The names Persicirhabdus sediminis gen. nov., sp. nov. (type strain YM20-087T =MBIC08313T =KCTC 22039T), Roseibacillus ishigakijimensis gen. nov., sp. nov. (type strain MN1-741T =MBIC08315T =KCTC 12986T), Roseibacillus ponti sp. nov. (type strain YM27-120T =MBIC08316T =KCTC 12987T), Roseibacillus persicicus sp. nov. (type strain YM26-010T =MBIC08317T =KCTC 12988T), Luteolibacter pohnpeiensis gen. nov., sp. nov. (type strain A4T-83T =MBIC08322T =KCTC 22041T) and Luteolibacter algae sp. nov. (type strain A5J-41-2T =MBIC08320T =KCTC 22040T) are therefore proposed. Emended descriptions of the class Verrucomicrobiae, the order Verrucomicrobiales and the family Verrucomicrobiaceae are also presented. PMID- 18398210 TI - Bacteroides xylanisolvens sp. nov., a xylan-degrading bacterium isolated from human faeces. AB - During the course of a study on the xylan-degrading community from the human gut, six xylanolytic, Gram-negative, anaerobic rods were isolated from faecal samples. 16S rRNA gene sequence analysis showed that the isolates were closely related to each other (> or =99 % sequence similarity) and that they belonged to the genus Bacteroides. On the basis of 16S rRNA gene sequence similarity, representative strain XB1AT was most closely related to the type strains of Bacteroides ovatus (97.5 %), B. finegoldii (96.5 %) and B. thetaiotaomicron (95.5 %). DNA-DNA hybridization results revealed that strain XB1AT was distinct from its closest relative, B. ovatus. The DNA G+C content of strain XB1AT (42.8 mol%) and major fatty acid composition (anteiso-C15 : 0, 33.8 %) further supported its affiliation to the genus Bacteroides. The novel isolates degraded different types of xylan, and were also able to grow on a variety of carbohydrates. Unlike most other Bacteroides species isolated from the human gut, these isolates were not able to degrade starch. Other biochemical tests further demonstrated that strain XB1AT could be differentiated from the closest related Bacteroides species. Xylan and sugars were converted by strain XB1AT mainly into acetate, propionate and succinate. Based on physiological, phenotypic and phylogenetic data, the six novel strains are considered to represent a novel species of the genus Bacteroides, for which the name Bacteroides xylanisolvens sp. nov. is proposed. The type strain is XB1AT (=DSM 18836T =CCUG 53782T). PMID- 18398211 TI - Humibacter albus gen. nov., sp. nov., isolated from sewage sludge compost. AB - A bacterial strain isolated from sewage sludge compost, strain SC-083T, was characterized. The isolate was a motile, Gram-positive, short rod, forming coryneform V-shaped cells during the early stages of growth. The organism was strictly aerobic and able to grow between 22 and 36 degrees C and between pH 5.5 and 8.0. The predominant fatty acids were cyclohexyl-C17 : 0, anteiso-C17 : 0 and iso-C16 : 0, the major respiratory quinones were menaquinone 11 (MK-11) and 12 (MK-12), and the genomic DNA G+C content was 68 mol%. The peptidoglycan contained the diagnostic diamino acids ornithine and 2,4-diaminobutyric acid and was of acetyl type. The 16S rRNA gene sequence analysis indicated that this isolate belongs to the family Microbacteriaceae with the type strains of the species Leifsonia xyli (96 % gene sequence similarity), Leifsonia shinshuensis (96 %), Leifsonia naganoensis (95 %), Leifsonia aquatica (95 %), Agromyces ramosus (95 %) and Curtobacterium citreum (95 %) among the closest phylogenetic neighbours. The phylogenetic analysis and phenetic characteristics support the proposal of a new genus and a novel species, with the name Humibacter albus gen. nov., sp. nov. The type strain of Humibacter albus is SC-083T (=DSM 18994T =CCUG 54538T =LMG 23996T). PMID- 18398212 TI - Microterricola viridarii gen. nov., sp. nov., a new member of the family Microbacteriaceae. AB - Strain KV-677T, a Gram-positive, aerobic, motile, rod-shaped bacterium, was isolated from park soil in Tokyo, Japan, and characterized. It grew well at 15-30 degrees C on nutrient agar and colonies were pale yellow. The cell-wall peptidoglycan contained diaminobutyric acid, glycine, alanine and glutamic acid and the muramic acid acyl type was acetyl. The predominant menaquinone was MK-12. Mycolic acids were not detected. The DNA G+C content was 70 mol%. 16S rRNA gene sequence analysis revealed that strain KV-677T fell within the cluster of the family Microbacteriaceae and formed a separate lineage joining the genera Salinibacterium, Rhodoglobus, Subtercola and Agreia, showing 95.5-96.9 % sequence similarities with the type strains of the type species of the above genera. However, strain KV-677T clearly differed from these and other genera with relatively high sequence similarity in its chemotaxonomic characteristics. Therefore, it is proposed that strain KV-677T represents a novel species in a new genus, Microterricola viridarii gen. nov., sp. nov., in the family Microbacteriaceae. The type strain of Microterricola viridarii is KV-677T (=NRRL B-24538T =NBRC 102123T). PMID- 18398213 TI - Chryseobacterium bovis sp. nov., isolated from raw cow's milk. AB - Three Gram-negative, rod-shaped, oxidase-positive, aerobic, non-motile bacterial strains, designated H9T, H10 and H15, were isolated during a study on the diversity of culturable psychrotolerant bacteria in raw cow's milk. Comparisons of 16S rRNA gene sequences showed that the three strains were very closely related to each other (sequence similarities of 99.6-99.8 %). A polyphasic taxonomic study of the isolates resulted in their identification as members of the genus Chryseobacterium (family Flavobacteriaceae, phylum Bacteroidetes). The three strains showed < or =96.9 % sequence similarity with respect to the type strains of described Chryseobacterium species, indicating that H9T, H10 and H15 represent a novel species of the genus Chryseobacterium. The three strains grew at 7-37 degrees C (strain H10 grew at up to 41 degrees C), with 0-2.5 % NaCl and at pH 5.0-9.8. The dominant cellular fatty acids of strain H9T were 15 : 0 iso (38.9 %), 15 : 0 anteiso (15.6 %) and 17 : 0 iso 3-OH (12.7 %). Strain H10 also possessed 17 : 1 iso omega 9c (14.8 %) as a major fatty acid. On the basis of phenotypic properties and phylogenetic distinctiveness, the three milk isolates represent a novel species in the genus Chryseobacterium, for which the name Chryseobacterium bovis sp. nov. is proposed. The type strain is H9T (=LMG 24227T =DSM 19482T). PMID- 18398214 TI - Streptomyces griseus 45H, a producer of the extracellular autoregulator protein factor C, is a member of the species Streptomyces albidoflavus. AB - Streptomyces griseus strain 45H, isolated in 1960 during a mutagenesis programme on the industrial streptomycin producer S. griseus 52-1, encodes an extracellular, pleiotropic autoregulatory signalling protein, factor C, which stimulates sporulation of S. griseus 52-1 in submerged culture. The facC gene, which codes for factor C, is present in very few streptomycetes and is not present in S. griseus 52-1. Based on 16S rRNA gene sequencing and other molecular data, S. griseus 45H, the factor C producer, is here shown to be related to the original laboratory strain of Streptomyces flavofungini, which was being studied in the same laboratory in 1960, and to Streptomyces albidoflavus. Southern blotting revealed that three out of four independently isolated strains of S. albidoflavus possess facC. Both the original strain of S. flavofungini and S. griseus 45H are therefore identified as members of the species Streptomyces albidoflavus, and we propose that S. griseus 45H should be renamed Streptomyces albidoflavus 45H. PMID- 18398215 TI - Aspergillus uvarum sp. nov., an uniseriate black Aspergillus species isolated from grapes in Europe. AB - A novel species, Aspergillus uvarum sp. nov., is described within Aspergillus section Nigri. This species can be distinguished from other black aspergilli based on internal transcribed spacers (ITS), beta-tubulin and calmodulin gene sequences, by AFLP analysis and by extrolite profiles. Aspergillus uvarum sp. nov. isolates produced secalonic acid, common to other Aspergillus japonicus related taxa, and geodin, erdin and dihydrogeodin, which are not produced by any other black aspergilli. None of the isolates were found to produce ochratoxin A. The novel species is most closely related to two atypical strains of Aspergillus aculeatus, CBS 114.80 and CBS 620.78, and was isolated from grape berries in Portugal, Italy, France, Israel, Greece and Spain. The type strain of Aspergillus uvarum sp. nov. is IMI 388523T=CBS 127591T=ITEM 4834T=IBT26606T. PMID- 18398217 TI - Proposal of Goodfellowiella gen. nov. to replace the illegitimate genus name Goodfellowia Labeda and Kroppenstedt 2006. AB - The prokaryotic, generic name Goodfellowia Labeda and Kroppenstedt 2006 is illegitimate because it is a later homonym of the name Goodfellowia Hartert, 1903 (Animalia, Chordata, Aves, Passeriformes, Sturnidae) [Principle 2 of the Bacteriological Code (1990 Revision)]. A new genus name, Goodfellowiella, is therefore proposed for this taxon (Rule 54). This also requires that a new combination, Goodfellowiella coeruleoviolacea comb. nov., be proposed for the type species to replace the illegitimate combination Goodfellowia coeruleoviolacea (Preobrazhenskaya and Terekhova 1987) Labeda and Kroppenstedt 2006. PMID- 18398216 TI - Dipodascus tetrasporeus sp. nov., an ascosporogenous yeast isolated from deep-sea sediments in the Japan Trench. AB - Dipodascus tetrasporeus sp. nov. is described as a novel yeast species in the family Dipodascaceae to accommodate an isolate recovered from sediments collected on the deep-sea floor in the north-western Pacific Ocean. In the clade comprising the genera Dipodascus, Galactomyces and Geotrichum, this is the only species that forms asci that bear four ascospores. The ascospore is surrounded by an irregular exosporium wall, similar to what is observed in the genus Galactomyces, but they are released by rupture, which is characteristic of Dipodascus and not Galactomyces. D. tetrasporeus is remarkably divergent (>10 % difference) in its D1/D2 26S rDNA sequence from any other known species. Although maximum-likelihood analysis of combined 18S rDNA and D1/D2 26S rDNA sequences cannot elucidate a reliable position for this species, it was placed among Geotrichum carabidarum, Geotrichum cucujoidarum, Geotrichum fermentans and Geotrichum histeridarum, which also have morphological and physiological affinity with the species. The species is homothallic. The type strain of Dipodascus tetrasporeus sp. nov. is strain SY 277T (=NBRC 103136T =CBS 10071T). PMID- 18398220 TI - Accumulation of cholesterol precursors and plant sterols in human stenotic aortic valves. AB - The pathogenesis of aortic valve stenosis (AS) is characterized by the accumulation of LDL-derived cholesterol in the diseased valves. Since LDL particles also contain plant sterols, we investigated whether plant sterols accumulate in aortic valve lesions. Serum samples were collected from 82 patients with severe AS and from 12 control subjects. Aortic valves were obtained from a subpopulation of 21 AS patients undergoing valve surgery and from 10 controls. Serum and valvular total cholesterol and noncholesterol sterols were measured by gas-liquid chromatography. Noncholesterol sterols, including both cholesterol precursors and sterols reflecting cholesterol absorption, were detected in serum samples and aortic valves. The higher the ratios to cholesterol of the cholesterol precursors and absorption markers in serum, the higher their ratios in the stenotic aortic valves (r=0.74, P<0.001 for lathosterol and r=0.88, P<0.001 for campesterol). The valvular ratio to cholesterol of lathosterol correlated negatively with the aortic valve area (r= -0.47, P=0.045), suggesting attenuation of cholesterol synthesis with increasing severity of AS. The higher the absorption of cholesterol, the higher the plant sterol contents in stenotic aortic valves. These findings suggest that local accumulation of plant sterols and cholesterol precursors may participate in the pathobiology of aortic valve disease. PMID- 18398221 TI - Decreased iPLA2gamma expression induces lipid peroxidation and cell death and sensitizes cells to oxidant-induced apoptosis. AB - Our previous studies showed that renal proximal tubular cells (RPTC) express Ca(2+)-independent phospholipase A(2)gamma (iPLA(2)gamma) in endoplasmic reticulum (ER) and mitochondria and that iPLA(2)gamma prevents and/or repairs lipid peroxidation induced by oxidative stress. Our present studies determined the importance of iPLA(2)gamma in mitochondrial and cell function using an iPLA(2)gamma-specific small hairpin ribonucleic acid (shRNA) adenovirus. iPLA(2)gamma expression and activity were decreased in the ER by 24 h and in the mitochondria by 48 h compared with scrambled shRNA adenovirus-treated cells. Lipid peroxidation was elevated by 2-fold at 24 h and remained elevated through 72 h in cells with decreased iPLA(2)gamma. Using electrospray ionization-mass spectrometry, primarily phosphatidylcholines and phosphatidylethanolamines were increased in iPLA(2)gamma-shRNA-treated cells. At 48 h after exposure to the iPLA(2)gamma shRNA, uncoupled oxygen consumption was inhibited by 25% and apoptosis was observed at 72 and 96 h. RPTC with decreased iPLA(2)gamma expression underwent apoptosis when exposed to a nonlethal concentration of the oxidant tert-butyl hydroperoxide (TBHP). Exposure of control cells to a nonlethal concentration of TBHP induced iPLA(2)gamma expression in RPTC. These results suggest that iPLA(2)gamma is required for the prevention and repair of basal lipid peroxidation and the maintenance of mitochondrial function and viability, providing further evidence for a cytoprotective role for iPLA(2)gamma from oxidative stress. PMID- 18398223 TI - Candidate genes of the 5-lipoxygenase pathway in acute coronary syndrome: a pilot study. AB - PURPOSE: The purpose of this pilot study was to examine arachidonate 5 lipoxygenase (ALOX5) and ALOX5-activating protein (ALOX5AP) gene variations in patients with and without acute coronary syndrome (ACS). METHODOLOGY: Four and six single nucleotide polymorphisms spanning the ALOX5 and ALOX5AP genes, respectively, were genotyped in 19 non-Hispanic Caucasian patients with ACS and 27 controls. RESULTS: Presence of the common allele of rs9508835 (ALOX5AP) and the minor allele of rs2029253 (ALOX5) were associated with ACS. After adjustment for age, being a carrier of the rs9508835 common allele was associated with an increased risk of ACS (odds ratio = 2.86). RELEVANCE FOR NURSING PRACTICE: Through the inhibition of the ALOX5AP gene by downregulation of the leukotriene pathway, the risk of ACS may be decreased in individuals that carry susceptibility allele(s). Knowledge of the genetic basis of treatments that downregulate the leukotriene pathway may prove essential to the care of individuals with ACS. PMID- 18398222 TI - A focused microarray to study human mitochondrial and nuclear gene expression. AB - A focused microarray (huMITOchip) was developed to study alterations of human mitochondrial and nuclear gene expression in health and disease. The huMITOchip contains 4,774 probe sets identical to the Affymetrix U 133 plus 2.0 chip covering genes affecting mitochondrial, lipid, cytokine, apoptosis, and muscle function transcripts. Unlike other gene chips, the huMITOchip has 51 probe sets that interrogate 37 genes of the mitochondrial genome. The human mitochondrial gene chip was validated against the Affymetrix U133 plus 2.0 array using an in vitro system of CCL136 muscle cell line stimulated with or without interferon gamma (IFN-gamma). The 37 genes from the mtDNA demonstrated absolute gene expression levels ranging from 0.1 to 3,182. The comparison of the two gene chips yielded an excellent Pearson's correlation coefficient (r = 0.98). At least 17 probe sets were differentially expressed in response to IFN-gamma on both chips, with a high degree of concordance. This is the first report on the development of a focused oligonucleotide microarray containing genes of the mitochondrial genome. PMID- 18398224 TI - Apoptotic-related protein expression in the diaphragm and the effect of dopamine during inspiratory resistance loading. AB - Dopamine (DA) is a free radical scavenger that attenuates apoptosis. We studied the effects of normal saline (NS) and DA on diaphragm apoptotic protein expression following 60 min of inspiratory resistance loading in rats. We tested for 27 apoptotic-related proteins and found 12 in the diaphragm. Of the 12 proteins, superoxide dismutase copper zinc (SOD [CuZn]) and proprioceptive event related potential (PERP) were significantly higher in the DA group than in the NS and sham groups (p = .002, p = .007). DA group diaphragms had significantly greater expression of SOD (CuZn) than the NS (p = .005) and sham group diaphragms (p = .003). Likewise, the DA group had significantly greater expression of PERP than the NS group (p = .008). These results suggest that DA decreases diaphragm apoptosis through elevated expression of SOD (CuZn). The identification of 12 apoptotic-related proteins will assist investigators as they study diaphragm apoptosis. PMID- 18398225 TI - Autonomic tone in medical intensive care patients receiving mechanical ventilation and during a CPAP weaning trial. AB - Mechanical ventilator support and the resumption of spontaneous ventilation or weaning create significant alterations in alveolar and intrathoracic pressure that influence thoracic blood volume and flow. Compensatory autonomic tone alterations occur to ensure adequate tissue oxygen delivery, but autonomic responses may produce cardiovascular dysfunction with subsequent weaning failure. The authors describe autonomic responses of critically ill patients (n = 43) during a 24-hr period of mechanical ventilatory support and during the 24 hr that included their initial spontaneous breathing trial using continuous positive airway pressure. Nearly two thirds of these patients demonstrated abnormal autonomic function and this dysfunction was more severe in those patients who were unable to sustain spontaneous ventilation (n = 15). With further systematic study, autonomic responses may be useful in the identification of patients who are likely to develop cardiac dysfunction with the resumption of spontaneous breathing. PMID- 18398226 TI - Methotrexate-induced alterations in beta-oxidation correlate with cognitive abilities in children with acute lymphoblastic leukemia. AB - Treatment advances, including central nervous system (CNS) treatment with methotrexate, have led to significant gains in disease-free survival from childhood acute lymphoblastic leukemia (ALL). However, methotrexate has been associated with neurological problems such as declines in cognitive and academic abilities. The purpose of this study was to investigate methotrexate-induced changes in beta-oxidation in children with ALL receiving methotrexate for CNS treatment. Specific aims were to investigate effects of methotrexate on beta oxidation of the two most prevalent fatty acids (palmitic acid and stearic acid) in cerebrospinal fluid (CSF) samples and correlate the ratio of monounsaturation to saturation of these fatty acids with cognitive and academic abilities. The sample included 12 females and 14 males with low-risk (n = 7), standard-risk ( n = 13), or high-risk (n = 6) ALL. Mean age at diagnosis was 94.1 months (SD = 34.4). CSF samples were obtained in conjunction with diagnostic lumbar punctures; subsequent samples were obtained prior to intrathecal methotrexate administration during the induction, consolidation, and continuation phases of treatment. Fatty acids were analyzed by gas chromatography. Results showed a significant increase in the ratio of monounsaturation to saturation of both fatty acids, which was greatest during the most intensive phase of treatment. Ratios of monounsaturated to saturated fatty acids were negatively correlated with full-scale IQ, verbal IQ, and math calculations. Findings suggest that methotrexate alters beta oxidation and that the resulting increase in fatty acid monounsaturation is related to declines in some domains of cognitive ability. PMID- 18398227 TI - Efficacy of a novel procedure sheath and closure device during diagnostic catheterization: the multicenter randomized clinical trial of the FISH device. AB - BACKGROUND: The aim of vascular closure devices is to safely secure the arterial access site at the conclusion of catheterization procedures, thereby increasing patient comfort and decreasing time to hemostasis and ambulation. The FISH (femoral introducer sheath and hemostasis) device is novel in that the access sheath and closure component are incorporated onto the same system. METHODS: The FISH pivotal investigation was conducted at 8 catheterization laboratories throughout the United States. Eligible diagnostic patients were randomized (2 to 1) to the FISH device versus manual compression and assessed for time to hemostasis and time to ambulation. Half of the participants underwent ultrasonographic evaluation at 30-day follow up. Enrollment for an interventional cohort is ongoing and will be reported at a later date; however, the interventional patients enrolled to date were combined with the diagnostic patients to comprise the safety data of the trial. RESULTS: Overall, 191 patients were randomized to the FISH device and 106 patients to manual compression. Most patients received a 6 Fr sheath (approximately 70%), while the remaining patients received a 5 or 8 Fr sheath. Twenty-seven patients who received the FISH device were converted to manual compression due to anticipated suboptimal hemostasis. Among the diagnostic patients, the mean time to hemostasis was 8.9 minutes for the FISH device, compared to 17.2 minutes for manual compression (p < 0.0001). Similarly, the mean time to ambulation was 2.4 hours for the FISH device, compared to 4.3 hours for manual compression (p < 0.0001). Among the total cohort, there was 1 death and 1 episode of major access-site-related bleeding that required transfusion occurred in the FISH group (1.1%), compared to no serious adverse safety events in the manual compression group (p = 1.0). For the FISH group, there were 5 minor adverse safety events; 3 access-site hematomas and 2 pseudoaneurysms treated with thrombin injection, and in the manual compression group, there was 2 access-site hematomas and 1 pseudoaneurysm treated with thrombin injection (p = 1.0). CONCLUSION: Among diagnostic patients with good sheath placement and favorable femoral anatomy, the FISH device is superior in achieving time to hemostasis and ambulation compared to manual compression. At 30 days, there is no apparent difference in serious or minor adverse vascular events with the use of the FISH device. PMID- 18398228 TI - Vascular closure devices: have we caught the right FISH? PMID- 18398229 TI - Short- or long-term outcomes of coronary artery aneurysms occurring after directional coronary atherectomy. AB - Coronary artery aneurysm (CAA) often occurs after percutaneous coronary intervention, and it could be recognized more often in coronary intervention with directional coronary atherectomy (DCA). However, it has been uncertain and the natural history of CAA after DCA remains obscure. Thus, we examined the clinical course after DCA. This study included 792 lesions in which a follow-up angiogram was completed at mid- or long-term (3 months [mos.] or more than 1 year after DCA). The mean average of the angiographic follow-up period was 24.8 mos. (range 3-128 mos.), and clinical follow-up period was 45.6 mos. (range 3-144 mos.). CAA was defined as 1.5 > DCA site diameter/reference diameter by quantitative coronary angiography (QCA). CAAs were detected in 21 lesions (2.7%). There was no significant difference in the target lesion revascularization rate between CAA and non-CAA lesion (19.0% vs. 24.6%). More than twice as many follow-up coronary angiograms were performed in 15 lesions among 21 CAA lesions. The mean duration of the follow-up angiograms was 19.6 mos. There was no significant difference in lumen diameter between the value of QCA at first and final follow-up coronary angiography (3.9 +/- 0.9 vs. 4.0 +/- 0.8 mm). Acute coronary events or coronary perforations did not occur at all. This study showed the frequency of DCA-related CAA and revealed acceptable short- or long-term prognosis. DCA-related CAA had not been progressing during the follow-up period. We concluded that almost all CAAs should be managed conservatively. PMID- 18398230 TI - Urgent PCI in patients with stent thrombosis: an observational single-center study comparing thrombus aspiration and standard PCI. AB - OBJECTIVES: Stent thrombosis (ST) is a recognized complication of percutaneous coronary interventions (PCI). Due to the high thrombus burden present in ST, the use of thrombus aspiration (TA) may enhance the angiographic results by limiting distal embolization. METHODS: Consecutive patients undergoing urgent PCI due to ST were studied, consisting of two groups according to the type of intervention (standard PCI [SP] or TA). TIMI flow, corrected TIMI frame count (cTFC), myocardial blush grade (MBG), thrombus score (TS), Yip's adverse thrombus burden features, occurrence of angiographically evident distal embolization (DE) were evaluated offline. In-hospital outcomes were also recorded. RESULTS: Twenty-eight patients entered the study. Baseline clinical and angiographic features were similar between patients treated by SP (n = 12) and those treated by TA (n = 16). After PCI, a trend in favor of TA as compared to SP was observed for post-PCI TIMI 3 flow rate (67% vs. 89%; p = 0.10), final cTFC (31 +/- 35 vs. 17 +/- 21; p = 0.10) and MBG 2-3 rate (40% vs. 67%; p = 0.12). Accordingly, the rate of adverse angiographic outcome (post-PCI TIMI 0-1 flow and/or DE) was significantly lower in the TA as compared to SP: 5% (1 out of 18) vs. 33% (5 out of 15) respectively (p = 0.04). Throughout hospitalization, major adverse events occurred in 6% of TA patients vs. 33% of SP patients (p = 0.08). CONCLUSIONS: TA in patients with ST is safe and is associated with better angiographic outcomes compared to standard PCI. Larger studies are warranted to confirm these early observations. PMID- 18398231 TI - Stent thrombosis--a complication best avoided. PMID- 18398232 TI - A novel method of PercuSurge distal protection in a five French guiding catheter without an Export aspiration catheter. AB - BACKGROUND AND OBJECTIVES: Although the PercuSurge (PS) system is an efficient distal protection device, it has several limitations such as significant occlusion time causing myocardial ischemia and requirement of a large 7 or 8 Fr guiding catheter (GC). To address these problems, we developed a new method of use for the PS system using a deeply-engaged 5 Fr GC as an aspiration catheter instead of the Export aspiration catheter. METHODS: We studied the initial feasibility and safety of 14 patients treated with a new PS procedure compared to 18 patients treated with the conventional method. RESULTS: Clinical and demographic characteristics were similar between the two patient groups. Device success rate was also similar. The new method using a 5 Fr GC without the Export aspiration catheter significantly reduced fluoroscopic time, total procedure time, occlusion time and in-hospital costs. CONCLUSIONS: This method may be an important alternative procedure that compensates for the disadvantages associated with the conventional use of the PS system by taking advantage of the flexibility and versatility of a 5 Fr GC. PMID- 18398233 TI - Transcatheter device closure of atrial septal defects in patients older than 60 years of age: immediate and follow-up results. AB - Atrial septal defect (ASD) closure reduces symptoms and prevents ongoing congestive heart failure. However, little is known about device closure in the elderly (age > 60 years) and whether it is a safe and effective treatment. In this study, we report our results with ASD transcatheter closure in the elderly patient > 60 years of age using the Amplatzer Septal Occluder (ASO) device. METHODS: Between September 1999 and March 2007, a total of 41 patients (24 females and 17 males) who were > 60 years of age (range 62-87.2 years) and had indications for ASD closure (right ventricular enlargement, clinically symptomatic dyspnea, fatigue, palpitations, exercise intolerance, transient ischemic attacks and/or stroke) underwent an attempt of transcatheter device closure using the ASO. RESULTS: The median Qp/Qs ratio was 2.3 (range 1-7.5). The median mean pulmonary artery pressure was 26 mmHg (range 11-52 mmHg). The median size of the ASD as measured by intracardiac echocardiography (ICE) (n = 38) or transesophageal echocardiography (TEE) (n = 3) was 18.9 mm (range 8-40 mm), and the median balloon-stretched (stop-flow technique) diameter (n = 32) was 23.5 mm (range 12-40 mm). The procedure was successful in all patients. 44 devices were deployed in the 41 patients (3 patients received 2 devices each). A 6 mm fenestration was created in a 30 mm device that was placed in 2 patients with left ventricle diastolic dysfunction. The median fluoroscopy time was 10 minutes (range 4-24.8 minutes), and the median procedure time was 60 minutes (range 26 110 minutes). Complications encountered during or within 24 hours after the procedure included: hematoma at the site of catheter insertion in 4 patients, small pericardial effusion in 5, and in 1 patient, the pacemaker lead was dislodged requiring reimplantation. The complete closure rate as assessed by ICE or TEE immediately after the procedure was 82%. However, complete closure, as assessed by transthoracic echocardiography at 24 hours and 6-24 months postintervention, was 82% and 97%, respectively. Two patients did not return for follow up and 4 patients were known to have expired for reasons unrelated to their ASD closure. At a median interval of 6 months after closure, the right ventricular end-diastolic dimension decreased from 38.9 +/- 9 mm preprocedure to 26.6 +/- 7 mm (p < 0.001; n = 31) postprocedure, and 89% of the patients showed improvement in symptoms. On a median follow-up interval of 28 months (range 6-79 months), 1 patient was noted to have a femoral pseudoaneurysm. Patients were doing well with no major clinical events. CONCLUSION: In patients older than 60 years of age, device closure of ASDs is safe and an effective alternative to surgery. Right-heart remodeling can occur in patients > 60 years of age. PMID- 18398234 TI - The declining relevance of age in the treatment of atrial septal defects. PMID- 18398236 TI - B-type natriuretic peptide and serum unbound free fatty acid levels after contemporary percutaneous coronary intervention. AB - OBJECTIVES: To determine the frequency and timing of B-type natriuretic peptide (BNP) and unbound free fatty acid (FFAu) elevation after percutaneous coronary intervention (PCI). DESIGN AND METHODS: Blood samples were collected from 55 patients undergoing PCI within 1 hour prior to PCI, immediately after PCI, 6 hours and 18-24 hours after PCI, and were analyzed for BNP and FFAu. RESULTS: There was a trend toward a rise in BNP levels at 18-24 hours post-PCI (65 vs. 45 pg/ml; p = 0.056). FFAu levels rose immediately after PCI and returned to baseline by 6 hours postprocedure (2.0 nM pre-PCI, 6.4 nM immediately post-PCI, 1.9 nM 6 hours post-PCI, and 2.2 nM 18-24 hours post-PCI; p < 0.0001). BNP and FFAu levels were elevated post-PCI in 17% and 82% of cases. CONCLUSIONS: PCI using short inflation times and coronary stenting are associated with a trend toward increased BNP levels at 18-24 hours post-PCI and a transient significant rise in FFAu levels. PMID- 18398235 TI - Long-term outcomes after intracoronary Beta-irradiation for in-stent restenosis in bare-metal stents. AB - OBJECTIVE: We sought to characterize the long-term outcomes of patients undergoing intracoronary brachytherapy using Beta- irradiation (Beta-BT). BACKGROUND: Beta-BT is effective in reducing angiographic restenosis as well as target vessel revascularization (TVR) in patients with in-stent restenosis (ISR) after bare-metal stenting (BMS). METHODS: 81 consecutive patients undergoing Beta BT for ISR (irradiated length 32 [32-54] mm) after BMS in native vessels (n = 79) or saphenous vein grafts (n = 2) between 2001 and 2003 were followed. Major cardiac events (MACE), including cardiac death, nonfatal myocardial infarction (MI), and TVR occurring > 1 year or > 1 year were assessed 5.2 (4.4-5.6) years after the index procedure. RESULTS: During the entire follow-up period, the total MACE rate was 49.4%. Within the first year and at > 1 year, MACE rates were 25.9% and 23.5%, cardiac death occurred in 2.4% and 6.2%, and nonfatal MI in 6.2% and 12.3% for annual cardiac death/MI rates of 8.7% at < 1 year and 4.1% thereafter. TVR was required in 19% at < 1 year and in 16% of patients later on. The only independent predictor of MACE occurring < 1 year was an irradiated vessel length > 32 mm (odds ratio [OR] 2.73, 95% confidence interval [CI] 1.10-6.78; p = 0.03). The best, albeit not statistically significant, predictor of MACE occurring at > 1 year was the presence of diabetes mellitus (OR 2.49, 95% CI 0.94-6.57; p = 0.07). CONCLUSIONS: Patients undergoing Beta-BT for ISR after BMS carry a substantial risk of MACE also beyond the first year, with annual cardiac death and nonfatal MI rates of 1.5% and 2.9% up to 5 years postprocedure. PMID- 18398237 TI - Protection devices and thrombectomy for native coronary artery ST-elevation myocardial infarction. AB - Distal thrombus embolization during primary percutaneous coronary intervention occurs frequently and is associated with compromised long term outcomes. Apart from pharmacological agents, great interest has recently been shown in various mechanical devices aimed at either reducing the amount of thrombus present or preventing its distal migration. However, despite the intuitive appeal of such devices, their application remains uncertain given the results of emerging randomized trials. Data presently available show a significant discrepancy between softer clinical endpoints such as reperfusion markers and hard clinical outcomes. The purpose of this review is to summarize the current clinical evidence and provide guidelines for their use. PMID- 18398238 TI - Cardiac magnetic resonance and "augmented" right-to-left intracardiac shunting through a patent foramen ovale. AB - Platypnea-orthodeoxia is an uncommon condition characterized by the development of hypoxia and breathlessness in the upright posture, relieved by resuming a supine position. First described in 1969, the condition has since been associated with intracardiac and intrapulmonary shunts, liver disease and a host of other conditions. We report an unusual case of episodic breathlessness and hypoxia cured by percutaneous cardiac intervention and discuss the evolving role of cardiac magnetic resonance imaging in the demonstration of functional distortions of cardiac and mediastinal anatomy that may not be as well characterized by other available imaging modalities. PMID- 18398239 TI - My history of carotid angioplasty and stenting. PMID- 18398240 TI - Percutaneous transcatheter left atrial appendage exclusion in atrial fibrillation. AB - Stroke is one of the leading causes of mortality, morbidity and serious disability in the developed world. Atrial fibrillation (AF), one of the most common cardiac arrhythmias, is a well-known predisposing factor for stroke, raising the risk significantly. Oral anticoagulation with warfarin is currently the most effective therapy for stroke risk reduction; however, this therapy increases the risk of bleeding and is often underutilized, contraindicated, or when administered, often subtherapeutic. It has been documented that the left atrial appendage (LAA) is the main source of left atrial thrombus, especially in non-rheumatic AF. Therefore, LAA exclusion may reduce the risk of stroke in AF, and retrospective surgical data have demonstrated a reduced risk of embolic events if surgical LAA exclusion was also performed during mitral valve replacement. Recently, several less invasive percutaneous transcatheter techniques of LAA exclusion - the PLAATO device, the Watchman device, and the Amplatzer Septal Occluder - have been employed with initially encouraging results. There is currently an ongoing randomized trial comparing percutaneous LAA exclusion to long-term oral anticoagulation therapy. Until such data are available, however, oral anticoagulation should remain the standard of care for stroke prevention in patients with AF. PMID- 18398241 TI - State-of-the-art nonvascular interventions: mitral valvuloplasty. PMID- 18398242 TI - Malignancy: an unrecognized risk factor for coronary stent thrombosis? AB - Stent thrombosis is a potentially catastrophic complication of coronary artery stenting. There have been particular concerns about the incidence of stent thrombosis following insertion of drug-eluting stents. We report a series of cases in which stent thrombosis occurred in association with malignancy and describe the potential mechanisms behind such an association. We speculate that this association merits further investigation as it raises the possibility that known malignancy may be a risk factor for stent thrombosis and that unexplained stent thrombosis, particularly if recurrent, should stimulate a search for occult malignancy. PMID- 18398243 TI - Angioplasty, stenting and thrombectomy to correct left main coronary stem obstruction by a bioprosthetic aortic valve. AB - Iatrogenic ostial coronary stenosis is a rare life-threatening complication of aortic valve replacement, usually presenting after 2-6 months. Although it is rarely reported, its incidence has been estimated at 0.3-5%. The most likely mechanism is posttraumatic fibrous intimal proliferation caused by coronary ostia cannulation for direct cardioplegia. We report a unique case of early occurrence of thrombotic obstruction of the left main stem following aortic valve replacement and its interventional management. PMID- 18398244 TI - Percutaneous balloon valvuloplasty of coexisting mitral and tricuspid stenosis: single-wire, double-balloon technique. AB - BACKGROUND: Percutaneous transmitral commissurotomy (PTMC) was first described by Inoue as an alternative to surgical closed mitral commissurotomy for severe rheumatic mitral stenosis. Two techniques - single- and double-balloon methods and percutaneous metallic devices - have been described for mitral dilatation. The Multi-Track system, a single-wire, double-balloon device was introduced by Bonhoeffer for mitral valve dilatation. We describe a case of mitral and tricuspid valve dilatation employing this double-balloon technique. METHODS: A 25 year-old male with combined rheumatic mitral and tricuspid stenosis and severe pulmonary hypertension underwent simultaneous double-valve dilatation in a single setting using different sizes of the Multi-Track balloon catheter. RESULTS: The patient's transmitral gradient decreased from 15 to 2 mmHg and his mitral valve area increased from 1 cm(2) to 2.2 cm(2). Similarly, his tricuspid valve gradient dropped from 7 to 3 mmHg and his tricuspid valve area increased from 0.9 cm(2) to 3.0 cm(2). CONCLUSION: Percutaneous balloon valvuloplasty using the Multi-Track system for combined mitral and tricuspid stenosis is cost effective and simple to use. PMID- 18398245 TI - Successful recanalization of in-stent coronary chronic total occlusion by subintimal tracking. AB - Percutaneous coronary intervention (PCI) of a chronic total occlusion (CTO) caused by in-stent restenosis (ISR) is sometimes very difficult due to the presence of hard occlusive components that prevent wire passage. We report a case of CTO caused by ISR (ISR-CTO) in which the occlusion was crossed with the wire subintimally along the outside of the stent and was successfully re-stented. Subintimal tracking along the outside of the stent can be considered as another approach for PCI of ISR-CTO in cases where conventional approaches fail. PMID- 18398246 TI - Percutaneous coronary intervention in neurosurgical patients. PMID- 18398247 TI - Collateral circulation via a rare, anomalously arising right ventricular branch. AB - Variations in coronary anatomy are not uncommon. They are usually benign, but can be a cause of confusion to the angiographer and in most situations, have no clinical significance. In this report, we present an unusual case of a chronically occluded proximal right coronary artery with an anomalous separate right ventricular branch arising directly from the aortic root and serving as a large collateral to the area beyond the occlusion. To our knowledge, this is the first of such anatomic variations being reported. PMID- 18398248 TI - Frugal coronary angioplasty, still an option after 30 years. PMID- 18398249 TI - Proceedings of the 4th Black Forest Forum for Musculoskeletal Interactions. May 10-13, 2007. Castle Bad Liebenzell, Germany. PMID- 18398250 TI - Bone quality: Emperor's new clothes. PMID- 18398251 TI - Genetically modified mechanostats: implications for skeletal competence? PMID- 18398252 TI - Mechanostat function during skeletal development. PMID- 18398253 TI - Phenotypic characterization of mouse models for bone-related diseases in the German Mouse Clinic. PMID- 18398254 TI - Describing force-induced bone growth and adaptation by a mathematical model. AB - This work proposes a mathematical model that qualitative describes the process of mechanically force-induced bone growth and adaptation. The mathematical model includes osteocytes as the key interfacing layer connecting tissue, cellular and molecular signaling levels. Specifically, in the presence of an increase in the mechanical stimuli, osteocytes respond by mechano-transduction releasing the local factors nitric oxide (NO) and prostaglandin E(2) (PGE(2)). These local factors act as the signaling recruitment signals for bone cells progenitors and influence the coupling activity among osteoblasts and osteoclasts during the process of bone remodeling. The model is in agreement with qualitative observations found in the literature concerning the process of bone adaptation and the cellular interactions during a local bone remodeling cycle induced by mechanical stimulation. PMID- 18398255 TI - Phytoestrogenic compounds for prevention of steroid-associated osteonecrosis. PMID- 18398256 TI - Differences in zinc status, bone turnover and femoral head bone density and biomechanical properties between patients with osteoarthritis and osteoporosis. PMID- 18398257 TI - Could image analysis of pQCT scans provide additional information about bone strength? PMID- 18398258 TI - Bone markers and bone mineral density in Duchenne muscular dystrophy. PMID- 18398259 TI - Scaling and adjusting growth-related data and sex-differences in the muscle-bone relation: a perspective. PMID- 18398260 TI - Histomorphometric evaluation of skeletal development. PMID- 18398261 TI - Limb lengthening: contribution of dual energy X-ray absorptiometry. PMID- 18398262 TI - Muscle area, muscle mass and muscle power in a pre-pubertal and an elderly population. PMID- 18398263 TI - Pain and its relationship with muscle strength and proprioception in knee OA: results of an 8-week home exercise pilot study. AB - Muscle strength and proprioception deficits have been recognized in knee OA. Pain is the symptomatic hallmark of knee OA. Indirect evidence suggests that muscle strength and proprioception deficits may be interrelated and that pain may have a confounding influence on the measurement of these factors in knee OA. However, these relationships have never been clearly evaluated. Therefore, the purpose of this investigation was to investigate relationships between pain, muscle strength, and proprioception in subjects with knee OA before and after an 8-week home exercise program. This study evaluated thirty-eight subjects with knee OA. Subjects were taught standard quadriceps strengthening exercises that were to be performed daily at home. Pain, muscle strength, and proprioceptive function were measured at baseline and after 8 weeks of therapy. Significant improvements in pain (42%, p<0.001) and quadriceps muscle strength (30%, p<0.001) were noted. Significant indirect associations were observed between pain and both muscle strength (rho=-0.39, p=0.01) and proprioceptive acuity (rho=-0.35, p=0.03) at baseline. Changes in pain were directly associated with changes in muscle strength (rho=0.45, p=0.005) and proprioceptive acuity (rho=0.41, p=0.01) with exercise. The association of pain with both muscle strength and proprioception should prompt future studies to consider and adjust for the influence of pain on neuromuscular factors in knee OA. PMID- 18398264 TI - Cellular isolation, culture and characterization of the marrow sac cells in human tubular bone. AB - The goal of this study is to characterize the epithelioid-like human marrow sac cells that separate the myeloid and osteoblast populations in situ and to determine if they express osteoblast cytoplasmic markers. Tubular segments of femoral diaphyseal bone were obtained from healthy young (4-8 yr) male and female patients undergoing femoral shortening surgeries. The interface between bone and marrow was examined by scanning (SEM) and transmission electron microscopy (TEM). The marrow sac cells were isolated and cultured in a-MEM medium with and without dexamethasone, glycerophosphate, and ascorbic acid [DGPA]. Alkaline phosphatase (ALP), bone morphogenetic protein-2 (BMP-2) and osteocalcin were evaluated. In the SEM, the marrow sac presented a distinctive pattern of large overlapping cells. TEM studies showed that marrow sac was one or two cells thick, which were attenuated with elongated nuclei, few cellular organelles, and appeared to display intercellular gap junctions. In culture, the marrow sac cells stained positively for ALP and BMP-2, and their expression was enhanced two- to three- fold when the cells were grown in DGPA. DGPA did not enhance osteocalcin expression. The cells of the human marrow sac reside proximate to endosteal osteoblasts and express osteoblastic markers. It is possible that these stromal cells constitute an osteoprogenitor pool from which replacement osteoblasts are recruited, and that they are involved in normal bone formation and in bone diseases (e.g., osteoporosis and osteopenia). PMID- 18398266 TI - Biomechanics of leg deformity treatment. AB - At pre-school age, there is a possibility to treat severe varosity and/or valgosity of legs by orthoses. The three points force effect of the orthosis partially corrects the pathologic defect of the leg. If the orthosis is intermittently used for a long time the correction is permanent. Step by step correction of bone deformities are based on remodelling of growth epiphyses and bones that are caused by orthotic bending pre-stressing. According to Huter Volkmann's law, the oblique loading regulates the growth of epiphyseal plates of long bone into the direction of the pressure result and the bone remodelling process is started: it means that the bone grows at the tensile part of growth epiphyses more quickly than at pressure one and gradually eliminates the varosity and/or valgosity defect. The knowledge of stress values for starting of the bone remodelling process is principal for clinical praxis. The values of pre-stressing cannot be increased by starting the remodelling process from the ethical point of view but it can be judged on its starting according to the success of the treatment. The aim of this article is to study the bone, ligament stress state and deformations of successful treatment. Method and calculation algorithm of stress state and deformation that are necessary for the starting of the remodelling process at the knee region were verified on a group of eight patients that were fitted by orthoses with bending pre-stressing. The space models of the knee, femur and tibia were composed with the help of X-ray, CT and MRI scan. The calculation algorithm was implemented on a PC and the program can be easily applied at clinical praxis. PMID- 18398265 TI - Age and motor score predict osteoprotegerin level in chronic spinal cord injury. AB - OBJECTIVE: Individuals with spinal cord injury (SCI) develop a severe form of osteoporosis below the level of injury that is poorly understood. We conducted a preliminary investigation to assess whether circulating markers of bone turnover and circulating RANKL/OPG levels are related to the severity of SCI, aging, or to differences in mobility (i.e., walking or using a wheelchair). METHODS: Sixty four caucasian men >or=1.6 years since injury selected based on locomotive mode provided blood samples and completed a health questionnaire at the VA Boston Healthcare System from 10/2003 to 6/2005. Plasma sRANKL, osteoprotegerin (OPG), osteocalcin and carboxyterminal telopeptide of type I collagen (CTx) levels were determined. RESULTS: Increasing age was significantly associated with increased OPG and CTx. Injury severity was predictive of OPG levels, and adjusting for age, participants with cervical motor complete and ASIA C SCI (n=11) had significantly lower mean OPG (46.1 pg/ml) levels than others (63.4 pg/ml). Locomotive mode was not associated with differences in bone markers. CONCLUSIONS: Severe cervical spinal cord injury is associated with decreased circulating OPG levels placing these patients at risk for accelerated bone loss that appears unrelated to locomotive mode. PMID- 18398267 TI - Decline of specific peak jumping power with age in master runners. AB - BACKGROUND: It is difficult to disentangle the effects of pure ageing from those of disuse. Master athletes, however, provide an opportunity to assess the effects of ageing per se, as these people maintain high activity levels during ageing. METHODS: We examined 200 female and 295 male master runners over the age of 35 who participated at European and World master championships. Runners were grouped by short, middle and long distance disciplines. Besides a questionnaire about their sports activities, measurements of counter movement jumps on a ground reaction force plate were performed. Specific peak jump power was the main subject, i.e., maximum jump power per body mass. RESULTS: All discipline groups showed an age-related decline in specific jump power when performing counter movement jumps (p<0.001). Except for female long distance runners, the amount of decline was the same for all discipline groups (p<0.001 to p<0.01) for each gender. The results for female long distance runners was highly spread caused by the small number of participants with older age. CONCLUSIONS: Our data indicate a decline in specific jump power that is similar to that reported in previous studies. The novelty from our results is the comparison of intra-gender decline. We observed the same amount of decline for all runners participating in different running disciplines. PMID- 18398268 TI - Site specific bone adaptation response to mechanical loading. AB - Over 25 million Americans suffer from osteoporosis. Bone size and strength depends both upon the level of adaptation due to physical activity (applied load), and genetics. We hypothesized that bone adaptation to loads differs among mice breeds and bone sites. Forty-five adult female mice from three inbred strains (C57BL/6 [B6], C3H/HeJ [C3], and DBA/2J [D2]) were loaded at the right tibia and ulna in vivo with non-invasive loading devices. Each loading session consisted of 99 cycles at a force range that induced approximately 2000 microstrain (microepsilon) at the mid-shaft of the tibia (2.5 to 3.5 N force) and ulna (1.5 to 2 N force). The right and left ulnae and tibiae were collected and processed using protocols for histological undecalcified cortical bone slides. Standard histomorphometry techniques were used to quantify new bone formation. The histomorphometric variables include percentage mineralizing surface (%MS), mineral apposition rate (MAR), and bone formation rate (BFR). Net loading response [right-left limb] was compared between different breeds at tibial and ulnar sites using two-way ANOVA with repeated measures (p<0.05). Significant site differences in bone adaptation response were present within each breed (p<0.005). In all the three breeds, the tibiae showed greater percentage MS, MAR and BFR than the ulna at similar in vivo load or mechanical stimulus (strain). These data suggest that the bone formation due to loading is greater in the tibiae than the ulnae. Although, no significant breed-related differences were found in response to loading, the data show greater trends in tibial bone response in B6 mice as compared to D2 and C3 mice. Our data indicate that there are site-specific skeletal differences in bone adaptation response to similar mechanical stimulus. PMID- 18398269 TI - Comparison of T- and Z-score in identifying risk factors of osteoporosis in inflammatory bowel disease patients. AB - OBJECTIVE: Most studies have shown contradictory results regarding predictive factors of osteoporosis in inflammatory bowel disease (IBD). Since in these studies either T- or Z-scores has been used, our aim was to compare T- and Z score in identifying risk factors of osteoporosis in IBD patients. MATERIALS AND METHODS: Bone density was measured by dual X-ray absorptiometry (DXA) at L2-L4 of the spine and femoral neck in 122 patients. Twenty-two clinical parameters were recorded prior to DXA and evaluated by univariate and multivariate analysis. RESULTS: On multivariate analysis, cumulative steroid dose was a predictive factor of femoral neck T-score (p<0.001) and Z-score (p=0.001). Age was a predictive factor of femoral neck T-score (p<0.001). BMI was a predictive factor of femoral neck Z-score (p=0.03). None of the other 19 variables tested had any predictive value for bone density. Age >or=55 years was a risk factor of low femoral neck T-score (OR 5.08, 95% CI 1.90-13.57, p=0.001), as was cumulative dose of prednisolone >or=5 g (OR 3.41, 95% CI 1.50-7.73, p=0.004). CONCLUSIONS: There is a discordance of results depending on whether T- or Z-scores are used in analysis. Among 22 parameters, cumulative steroid dose and age proved to be the most important factors. PMID- 18398270 TI - An 18-month-old girl with a history of convulsions and a facial nevus. PMID- 18398271 TI - A case of hypocortisolemic clinical Cushing's syndrome. PMID- 18398273 TI - Skin tags as a presenting sign of basal cell nevus syndrome in three sisters of the same family. PMID- 18398272 TI - Ovarian hemangioma occurring synchronously with serous papillary carcinoma of the ovary and benign endometrial polyp. PMID- 18398274 TI - Plasma homocysteine concentrations and serum lipid profile as atherosclerotic risk factors in subclinical hypothyroidism. AB - BACKGROUND AND OBJECTIVES: Because subclinical thyroid dysfunction may be a risk factor for cardiovascular disease, we evaluated the atherosclerosis tendency in subclinical hypothyroid (SCH) patients. PATIENTS AND METHODS: Fifty-three subclinical hypothyroid patients (serum thyrotropin [TSH] concentrations >4.12 mU/L) were compared with a control group of 50 euthyroid subjects whose age, sex and body mass indices were similar to the patient group. We tested whether serum TSH concentrations were correlated with plasma total homocysteine concentration (tHcy), low-density lipoprotein cholesterol (LDL-C), very low-density lipoprotein cholesterol (VLDL-C), high-density lipoprotein cholesterol (HDL-C), total cholesterol (TC) and triglycerides (TG). RESULTS: There was a significant statistical difference between the patient and control groups for normal free T4 (1.02+/-0.17 vs. 0.86+/-0.13, P<.001), TSH (1.64+/-1.02 vs. 6.62+/-2.61, P<.001), TC (185+/-39 vs. 206 +/- 42, P=.01), TG (103+/-54 vs. 132+/-85, P=.04), LDL-C (114+/-33 vs. 127+/-36, P=.04), and TC/HDL-C (3.81+/-106 vs. 4.19+/-1.02, P=.04), respectively. No statistically significant difference was found between the two groups for HDL-C, VLDLC, LDL-C/HDL-C, and tHcy. Serum TSH was significantly correlated with plasma tHcy (r=0.55; P=.001), TC (r=0.52; P=.001), LDL-C (r=0.49; P=.001), TC/HDL-C (r=0.38; P=.002) and LDL-C/HDL-C (r=0.36; P=.004) across all participants. CONCLUSION: Our study suggests that the atherogenicity of SCH is not mediated by hyperhomocysteinemia. Associated hyperlipidemia may explain the observed increased risk of coronary artery disease in patients with SCH. PMID- 18398275 TI - Melkersson-Rosenthal syndrome with partial oculomotor nerve palsy. PMID- 18398276 TI - Percutaneous vertebroplasty in patients with osteoportic vertebral body fractures. PMID- 18398277 TI - Seasonal variation of appendicitis in northern Saudi Arabia. PMID- 18398278 TI - Tracheal intubation without muscle relaxants: a randomized study of remifentanil or alfentanil in combination with thiopental. AB - BACKGROUND AND OBJECTIVE: The combination of alfentanil-propofol or remifentanil propofol provides adequate conditions for tracheal intubation without neuromuscular blocking drugs in most patients, but hypottension can occur during induction of anesthesia with propofol. We compared clinically acceptable intubating conditions and cardiovascular responses to induction and endotracheal intubation in patients receiving either alfentanil 40 microg/kg or remifentanil 2, 3 or 4 microg/kg, followed by thiopental 5 mg/kg. PATIENTS AND METHODS: In a randomized trial, 80 patients were assigned in equal numbers to one of four groups: remifentanil 2, 3, or 4 microg/kg (groups R2, R3, R4, respectively) or alfentanil 40 microg/kg (group A40). In each group, the injection was given over 90 seconds followed by thiopental 5 mg/kg. Ninety seconds after the administration of thiopental, laryngoscopy and intubation were attempted. Intubating conditions were assessed as excellent, satisfactory, fair, or unsatisfactory. Arterial blood pressure and heart rate changes accompanying the four induction techniques were also recorded. RESULTS: Overall conditions at intubation were significantly better (P<.05), and the frequency of excellent conditions was significantly higher (P<.05) in the R4 or A40 group compared with the R2 or R3 group. Intubating conditions were not significantly different between group R4 and A40 (P>.05). The highest dose of remifentanil (4 microg/kg) resulted in an 18.7% decrease in mean arterial pressure (MAP) after induction of anesthesia compared with a 16.4% decrease in MAP with alfentanil 40 microg/kg (difference not statistically significant). CONCLUSION: The administration of remifentanil 4 I(1/4)g/kg or alfentanil 40 microg/kg before thiopental 5 mg/kg provided good to excellent conditions for endotracheal intubation with acceptable hemodynamic changes. PMID- 18398279 TI - CA-125: a marker for diagnosis and follow-up of pleuroperitoneal and lymph node tuberculosis. PMID- 18398280 TI - Drug-eluting stents: insights into safety and indications. PMID- 18398281 TI - The current status of medical education in the Gulf Cooperation Council countries. AB - BACKGROUND AND OBJECTIVES: In the last two decades, the curriculum in medical education has changed so as to maintain its efficiency and effectiveness. Considerable changes are underway in many medical colleges worldwide. This study assessed the current status of the undergraduate curricula in the medical colleges of the Gulf Cooperative Council (GCC) countries, in relation to the SPICES (student-centered, problem-based, integrated, community-based, elective and systematic) model. METHODS: A structured open-ended data form was used in the collection of information from the appropriate authorities in 30 medical colleges of the GCC countries (Saudi Arabia, Oman, Kuwait, Qatar, Bahrain, United Arab Emirates and Yemen) in the year 2005. RESULTS: Of 30 medical colleges, 13 (43.3%) were located in Saudi Arabia. The annual intake of students in the year 2005 in these 30 colleges was 3225, of which 64.15% were males. Twelve colleges (40%) followed the traditional curriculum, while the remaining (60%) followed a hybrid problem-based learning (PBL) curricula. Most of the colleges were moving towards the more desirable aspects of the SPICES model. The majority of the traditional colleges were planning to change their curricula to hybrid PBL curricula. Almost all new medical colleges were adopting the hybrid PBL curricula. CONCLUSION: Despite the diversity in the curricula followed in medical colleges in GCC countries, most of these colleges either are following or are moving towards the new trends in medical education curricula. PMID- 18398282 TI - Impact of urbanization on the prevalence and pattern of arterial hypertension on the island of Socotra. PMID- 18398283 TI - Neurobrucellosis mimicking demyelinizating disorders. PMID- 18398284 TI - The cardiovascular risks of etoricoxib (Arcoxia). PMID- 18398285 TI - Increased bone mineral density in postmenopausal women with type 2 diabetes mellitus. AB - BACKGROUND AND OBJECTIVES: Studies of bone mineral density (BMD) in women with type 2 diabetes mellitus have shown conflicting results. We conducted this study to determine whether postmenopausal women with diabetes have higher BMD than non diabetic women of similar age, and to investigate the relationship between BMD and relevant clinical characteristics in these groups of women. PATIENTS AND METHODS: We retrospectively analyzed lumbar spine, femoral neck, and radius BMD data and other relevant clinical data for 130 postmenopausal women with type 2 diabetes mellitus and 166 non-diabetic women collected during a voluntary screening for osteoporosis in postmenopausal women without a history of low bone mass or osteoporotic fractures. RESULTS: Women with type 2 diabetes mellitus had significantly higher mean lumbar spine BMD ( 0.903+/-0.165 vs. 0.824+/-0.199, respectively, P<.001) and mean femoral neck BMD (0.870+/-0.132 vs. 0.832+/-0.134, respectively, P<.05) than non-diabetic women. In both groups of women, age correlated negatively with BMD levels at all three anatomical sites. Higher body mass index was associated only with higher lumbar spine BMD in both groups. Alkaline phosphatase levels showed a negative correlation with BMD at all sites in women with type 2 diabetes mellitus. CONCLUSION: Postmenopausal women with type 2 diabetes mellitus have higher BMD levels than non-diabetic women with similar clinical characteristics, and require a more scrutinized approach in managing low bone mass. PMID- 18398286 TI - Surgical procedures performed in the neonatal intensive care unit on critically ill neonates: feasibility and safety. AB - BACKGROUND AND OBJECTIVE: Transferring unstable, ill neonates to and from the operating room carries significant risks and can lead to morbidity. We report on our experience in performing certain procedures in critically ill neonates in the neonatal intensive care unit (NICU). We examined the feasibility and safety of such an approach. METHODS: All surgical procedures performed in the the NICU between January 1999 and December 2005 were analyzed in terms of demographic data, diagnosis, preoperative stability of the patient, procedures performed, complications and outcome. Operations were performed at bedside in the NICU in critically ill, unstable neonates who needed emergency surgery, in neonates of very low birth weight (<1000 g) and in neonates on special equipment like high frequency ventilators and nitrous oxide. RESULTS: Thirty-seven surgical procedures were performed including 12 laparotomies, bowel resections and stomies, 7 repairs of congenital diaphragmatic hernias, 4 ligations of patent ductus arteriosus, and various others. Birthweights ranged between 850 g and 3500 g (mean, 2000 g). Gestational age ranged between 25 to 42 weeks (mean, 33 weeks). Age at surgery was between 1 to 30 days (mean, 10 days). Preoperatively, 19 patients (51.3%) were on inotropic support and all were intubated and mechanically ventilated. There was no mortality related to surgical procedures. Postoperatively, one patient developed wound infection and disruption. CONCLUSION: Performing major surgical procedures in the the NICU is both feasible and safe. It is useful in very low birth weight, critically ill neonates who have a definite risk attached to transfer to the operating room. No special area is needed in the the NICU to perform complication-free surgery, but designing an operating room within the the NICU would be ideal. PMID- 18398287 TI - Outcome of surgery for intramedullary spinal ependymoma. AB - BACKGROUND AND OBJECTIVES: Ependymoma is the most frequently encountered intramedullary tumor. Total surgical resection is the therapeutic modality of choice whenever possible, but carries a significant risk of morbidity. This study was designed to define prognostic factors that affect clinical outcome after surgical resection of spinal intramedullary ependymoma. PATIENTS AND METHODS: The medical records, radiological and pathological studies of all patients with intramedullary spinal ependymomas treated surgically in one institution were reviewed retrospectively. Spinal myxopapillary ependymomas were excluded. In a multivariable regression analysis, possible prognostic factors were correlated with the 6-month postoperative neurological status using McCormick's grading scale. RESULTS: Surgery was performed on 17 patients (14 males, 3 females, mean age of 42+/-15 years) with spinal ependymoma. The cervical spine was the most common tumor location (71%). Total surgical resection of the tumor was achieved in 11 cases (65%). Intraoperative neurophysiological monitoring was used in 8 cases (47%). Postoperatively, 11 patients (65%) either improved or had no change from their preoperative neurological status. None of the 11 totally resected tumors has shown evidence of recurrence in a follow-up period range from 8-120 months (median, 33 months). Of several possible prognostic factors, the only statistically significant correlation identified with the 6-month postoperative neurological status was the preoperative McCormick grading score. CONCLUSIONS: Preoperative neurological status was the only statistically significant factor in determining the postoperative neurological outcome of patients with spinal intramedullary ependymomas. Early diagnosis and referral for surgery to specialized centers are recommended as controllable factors in improving outcome. PMID- 18398289 TI - Re: Ibn al-Nafis: discoverer of the pulmonary circulation. PMID- 18398288 TI - Ibn Rushd (Averroes): Prince of Science. PMID- 18398291 TI - Autophagy in hematopoietic stem/progenitor cells exposed to heavy metals: Biological implications and toxicological relevance. AB - The inherent toxicity of many metal compounds, together with their widespread environmental distribution, raises concerns of potential health hazards. Little is known about the impact of these important environmental toxicants on adult stem/progenitor cells, necessary for tissue homeostasis and repair. We recently reported that autophagy is implicated in the response of hematopoietic stem/progenitor cells to toxic concentrations of hexavalent chromium (Cr[VI]) and cadmium (Cd), two well known carcinogenic heavy metal cations. Autophagy may lead to cell death if carried out too extensively, but also acts as a survival pathway in cells under stress. In stem/progenitor cells, an autophagic phenotype could mitigate metal-induced toxicity, contributing to the conservation of tissue renewal capability. Given the key role of toxic damage to adult stem/progenitor cells in cancer, it is necessary to investigate whether autophagic responses modulate the carcinogenic potential of exposure to heavy metals during stem/progenitor cell differentiation. PMID- 18398292 TI - The Ubi brothers reunited. AB - Atg12 and Atg8/LC3 are two ubiquitin-like proteins involved in autophagosome formation. They show several similar characteristics just like brothers evolved from the same ancestor, however, their functional relationship has been obscure. We recently reported that a super protein complex, the Atg16L complex, which consists of multiple Atg12-Atg5 conjugates and the associating protein Atg16L, has an E3-like role in the LC3 lipidation reaction(1). The activated intermediate, LC3-Atg3 (E2) is recruited to the site where the lipidation takes place by virtue of the Atg16L complex. Thus, these two closely resembling systems are connected also in terms of their functions. This finding will provide further important clues as to the origin of the autophagosome membrane, and how the process is regulated by starvation and PtdIns3P signals. PMID- 18398293 TI - Mitochondrially localized EGFR is subjected to autophagic regulation and implicated in cell survival. AB - Although generally acknowledged as a plasma membrane protein, the epidermal growth factor (EGF) receptor has been found in the nucleus and subcellular organelles. Recently, the mitochondrial localization of the EGF receptor (EGFR) was reported; nevertheless, the molecular mechanism underlying EGFR localization in mitochondria is largely unknown. Using immunofluorescence and immunoelectron microscopy, we observed that EGFR did localize within mitochondria. Moreover, EGFR mitochondrial translocation can be increased by rapamycin treatment in A431 cells and greatly reduced by the presence of 3-methyladenine (3-MA), an inhibitor of autophagy. The reduction of mitochondrial EGFR via autophagy inhibition is further confirmed by small interference RNA (siRNA), through which the essential protein Beclin 1 was depleted. Knocking down Beclin 1 markedly decreased the mitochondrial translocation of EGFR that was induced by rapamycin. We also noticed that the content of mitochondrial EGFR transfer is decreased when the cells are exposed to the apoptotic inducer etoposide. Additionally, either EGF treatment or EGFR knockdown by siRNA results in a greater decline of cell viability in cells possessing more mitochondrial EGFRs. Taken together, we conclude that EGFR mitochondrial localization is regulated by either autophagy or programmed cell death and is correlated with cell survival. PMID- 18398294 TI - A multipronged approach to prostate cancer. PMID- 18398295 TI - Contrast enhanced MRI and intravital fluorescence microscopy indicate improved tumor microcirculation in highly vascularized melanomas upon short-term anti VEGFR treatment. AB - Anti-angiogenic therapy by blocking VEGF signalling combined with standard chemotherapy is a novel strategy for clinical cancer treatment. The mechanisms for enhanced antitumoral effects are still a matter of controversial debate. Tumor vessel "normalization" upon anti-angiogenic therapy leading to improved drug delivery has been proposed as possible mechanism. Therefore, aim of the study was to investigate tumor microvascular function upon anti-VEGFR treatment in highly vascularized melanomas. A detailed intravital-microscopic analysis of tumor microcirculation including the distribution pattern of vessel diameters and blood flow velocities was performed in melanomas grown in dorsal skinfold chambers of hamsters. Animals with highly vascularized established tumors were treated by a VEGFR tyrosin kinase inhibitor (SU5416) on 3 repetitive days. Tumor tissue oxygenation was measured by phosphorescence quenching technique. Overall tumor microcirculation of subcutaneous tumors was investigated by contrast enhanced MRI (CE-MRI). Vessel density was significantly decreased in treated animals. A significant shift in the distribution patterns towards increased vessel diameters and faster red blood cell velocities in remaining tumor vessels was observed upon anti-VEGF treatment, compensating reduced vascular density. Moreover, a trend towards elevated pO(2) levels in treated tumors was observed. Compared to controls, inflow kinetics of tumors quantified by CE-MRI as well as overall uptake of contrast agent in tumor tissue were significantly increased following short-term SU5416 treatment. In conclusion the results confirm temporarily improved tumor microvascular function in highly vascularized melanomas upon short term anti-VEGFR treatment leading to enhanced tumor blood supply and oxygenation potentially improving the efficacy of simultaneous chemo- or radiotherapy. PMID- 18398296 TI - Protective immunity following vaccination: how is it defined? AB - Vaccination represents an important medical breakthrough pioneered by Edward Jenner over 200 years ago when he developed the world's first vaccine against smallpox. To this day, vaccination remains the most effective means available for combating infectious disease. There are currently over 20 vaccines licensed for use within the US with many more vaccines in the R&D pipeline. Although vaccines must demonstrate clinical efficacy in order to receive FDA approval, the correlates of immunity vary remarkably between different vaccines and may be based primarily on animal studies, clinical evidence, or a combination of these sources of information. Correlates of protection are critical for measuring vaccine efficacy but researchers should know the history and limitations of these values. As vaccine technologies advance, the way in which we measure and define protective correlates may need to evolve as well. Here, we describe the correlates of protective immunity for vaccines against smallpox, tetanus, yellow fever and measles and compare these to a more recently introduced vaccine against varicella zoster virus, wherein a strict correlate of immunity has yet to be fully defined. PMID- 18398297 TI - Differences in vaccinations in European Union. AB - The European Union (EU) currently has 27 Member States, each with its own history, characteristics and habits. The National Health Services of most of these countries have different vaccination systems, different vaccine recommendations and different schedules of vaccine administration, which means that immunization is not considered in the same way and, at least for some antigens, vaccination coverage does not always meet changing medical needs. Together with a lack of political will concerning prevention in childhood, a poor understanding or false perceptions on the part of the general public (and even healthcare workers), and the inadequacies and heterogeneity of the vaccination systems can all be considered barriers to vaccinations in Europe. The most important limitations are those relating to the evaluation of vaccination coverage, the lack of active reminder systems to pick up patients who miss appointments, and the monitoring of adverse events. A common programme designed to overcome these limitations could be beneficial in promoting vaccinations everywhere, above all because active measures by the Health Authorities to demonstrate the importance they attribute to vaccination could convince still uncertain parents to have their children vaccinated. PMID- 18398298 TI - Intradermal recombinant hepatitis B vaccination (IDRV) for non-responsive healthcare workers (HCWs). AB - This was an observational study of non-responsive HCWs who were referred to Stockport Specialist Immunisation Clinic (SSIC) between 1(st) January 2003 and 31(st) May 2006. Anti-HBs titres were determined 4-6 weeks after each intradermal recombinant vaccine (IDRV). Median anti-HBs titres were compared using the exact Wilcoxon rank sum test. In total, 23 eligible non-responding HCWs were identified. Protective anti-HBs titres (> or =10 mlU/ml) were induced in the majority of non-responders [21/23 (91.3%)] following two doses of IDRVs. HCWs who responded to the 1(st) IDRV with anti-HBs levels > or =10 mlU/ml were significantly younger and received their 1(st) IDRV more than six months after the last IM dose. Two HCWs (41 and 45 year old females) anti-HBs titres remained below 10 mlU/ml even after a 3(rd) dose. Anti-HBs titres were available for 40% (9/23) of the HCWs six or more months after the last maximum anti-HBs titres were achieved. None of the nine HCWs anti-HBs titres declined to less than 10 mlU/ml six or more months after the last maximum anti-HBs titres were achieved. However, a larger study with long-term follow-up is needed to determine the duration of protection following IDRV. HCWs with anti-HBs titres <10 mlU/ml after two full courses of intramuscular recombinant vaccine (IMRV) should be offered two doses of IDRVs followed by assessment of anti-HBs titres one to four months after the second dose in order to identify persistent non-responders. No significant adverse events were noted with IDRV being well tolerated and acceptable to HCWs. PMID- 18398299 TI - Attitudes towards vaccines and infectious disease risk among U.S. troops. PMID- 18398300 TI - The structure and function of immunization advisory committees in Western Europe. AB - National immunization advisory committees are charged with making recommendations to either a governmental agency or the public regarding which vaccines that children and/or adults at specific ages, or with specific risk factors, should receive. However, there may be significant variability in a variety of aspects of such committees including the manner in which they are constituted, the process of their deliberations, and their authority. We explored the policies and roles of immunization advisory committees in each of ten of the largest nations in Western Europe and found significant variation among the nations. Conceptually, this variation was able to be categorized into six specific domains; committee membership, processes of meetings, basis for decision making, financial issues, authority and the role of manufacturers and insurers. PMID- 18398301 TI - Seroprevalence of anti Vi antibodies and immunogenicity of Typhim Vi vaccine in children. AB - This prospective study was carried out on 250 children between 6 months to 5 years of age to determine seroprevalence of anti Vi antibodies and to measure seroresponse and percent seroconversion to TyphimVi polysaccharide vaccine in children 2-5 years of age. Fifty children each were enrolled between 6 to 12 months of age (Group A), between 1- 2 years of age(Group B), between 2-3 years of age (Group C), between 3-4 years of age (Group D) and between 4-5 years of age (Group E). Anti-Vi antibody baseline titres were determined in all children. Children in Groups C to E were vaccinated with Typhim Vi vaccine. Baseline and postvaccination antibody titres were determined by ELISA. Test sera which had antibody levels >1 microg/ml were scored as seropositive. Of 250 children, 3 had base line anti-Vi antibodies >1 microg/ml. Following immunization overall seroconversion rate was 77.5% with 65.3%, 78.2% and 88% children showing seroconversion in Groups C, D and E respectively. Seroconversion was significantly more in Group E children compared to Group C (p=0.0148). There were no significant adverse reactions following vaccination. The study highlights very low prevalence of baseline anti Vi antibodies in children between 6 months and less than 5 years of age and shows high immunogenicity and safety of Typhim Vi polysaccharide vaccine in children 2-5 years of age. PMID- 18398302 TI - Sustaining immunity after immunization against encapsulated bacteria. AB - Infections by encapsulated bacteria are important causes of infant mortality worldwide. Over the last 20 years protein-polysaccharide conjugate vaccines have been developed to protect against the major invasive bacterial diseases of childhood, Streptococcus pneumoniae, Haemophilus influenzae type b (Hib) and Neisseria meningitidis. These vaccines are highly immunogenic and have resulted in a huge reduction in the diseases caused by these bacteria in the countries that have introduced them in their immunisation schedules. However, it has been reported that infant immunisation is associated with a relatively short duration of antibody levels and vaccine effectiveness, despite the demonstrable presence of booster responses to further vaccine dose. In contrast, at older ages, more sustained protection has been described with just a single dose of a conjugate vaccine. Understanding the generation of long-term immunity, by protein polysaccharide conjugate vaccines, is essential to reduce infant mortality through the improvement of vaccine formulation and scheduling. PMID- 18398303 TI - Improving vaccine delivery using novel adjuvant systems. AB - Adjuvants have been common additions to vaccines to help facilitate vaccine delivery. With advancements in vaccine technology, several adjuvants which activate immune specific responses have emerged. Available data show these adjuvants elicit important immune responses in both healthy and immunocompromised populations, as well as the elderly. Guidelines for the use and licensure of vaccine adjuvants remain under discussion. However, there is a greater understanding of the innate and adaptive immune response, and the realization of the need for immune specific adjuvants appears to be growing. This is a focused review of four adjuvants currently in clinical trial development: ASO4, ASO2A, CPG 7907, and GM-CSF. The vaccines including these adjuvants are highly relevant today, and are expected to reduce the disease burden of cervical cancer, hepatitis B and malaria. PMID- 18398304 TI - Functional activity of antisera to group B streptococcal conjugate vaccines measured with an opsonophagocytosis assay and HL-60 effector cells. AB - Conjugate vaccines against group B Streptococcus (GBS), which is a leading cause of bacterial disease among newborns and the elderly with underlying illnesses, have progressed from animal studies to phase 1 and 2 clinical trials in healthy adults. Due to the wide-spread use of antibiotics to treat at-risk deliveries, a phase 3 efficacy trial of a GBS vaccine to prevent neonatal disease in the United States is unlikely. A viable approach to assess a vaccine's efficacy is to use a surrogate of protection which in the case of GBS is the opsonizing activity of serum antibody. The opsonophagocytosis assay (OPA) measures the ability of serum antibody to opsonize GBS for killing by effector cells in the presence of complement. In this report we demonstrate that differentiated HL-60 cells can substitute for human peripheral blood leukocytes (hPMNLs) in the OPA. Antisera to GBS type Ia CPS and type III CPS conjugate vaccines opsonized homologous GBS for killing at effector cells to GBS ratios of 2-4:1 regardless of whether HL-60 or hPMNLs were used. These results represent the first important step in developing a standardized, high-throughput OPA that could be used to assess the functional activity of vaccine-induced antibody and potentially serve as a surrogate of efficacy. PMID- 18398305 TI - Predicting response to HBV vaccination in people with positive anti-HBc but negative HBsAg and anti-HBs. AB - OBJECTIVE: There are 5.1-6.5% of people with positive anti-HBc in Iran. The aim of this study was to assess the predicting factors of response to hepatitis B vaccination in anti-HBc positive subjects. RESULTS: Total response rate to vaccination was 79.8% (75 cases) and 67.9% (38 cases) in cases and controls, respectively. Nineteen persons (20.2%) in cases and 18 persons (32.1%) of controls had negative anti-HBs even after three doses of HB vaccination. Factor associated with decreased response to vaccination was prior history of being HBsAg positive (OR = 1.3, p = 0.01). METHODS: In a quasi-experimental study, 94 people with negative HBsAg, negative anti-HBs and positive anti-HBc (cases) and 56 persons with negative HBsAg, anti-HBs and anti-HBc (controls) were vaccinated at zero, one and six months with recombinant hepatitis B vaccine. Successful immunization was defined by anti-HBs antibody titer > or =10 mIU/mL. CONCLUSION: The rate of response to hepatitis B vaccination is nearly like other studies but somewhat different. Higher percent of married cases together with higher percent of positive HBsAg in spouses may explain the slight difference in the response to vaccination in cases in comparison with controls as a result of booster like effect that seldom happens because of recurrent contacts between the subjects and the HBsAg positive spouses spontaneously. PMID- 18398306 TI - Development of a sandwich enzyme-linked immunosorbent assay (ELISA) for determining of bovine serum albumin (BSA) in trivalent measles-mump-rubella (MMR) vaccines. AB - A sandwich enzyme-linked immunosorbent assay (ELISA), using polyclonal antibody, was developed and compared with the commercial kit for detecting and estimating of BSA content in Measles-Mump-Rubella (MMR) vaccine samples in detection limit of nanogram level. The test depends on the capturing and detecting of BSA antigen by the polyclonal antibody. Initially, a detection range of 0-64 ng/ml was established, could be used for estimation of BSA content according to WHO requirement (50 ng/ml) in MMR vaccines. Comparative analysis of the test results for 85 MMR vaccine samples obtained with the commercial kit gave a sensitivity of 58.8% and a specificity of 97%. A high correlation (r = 0.94) was observed between BSA sandwich ELISA and commercial kit for BSA content in MMR samples. However, variations in values also were observed for the two assays. These variations may have been due to difference of upper limit of detection range of BSA content in commercial kit (32 ng/ml) and new sandwich ELISA (64 ng/ml) as well as the use of a different polyclonal antibody. In concerning the cutoff value for the WHO requirement and employment of standard solution of 64 ng/ml in developing assay, it would be adequate to use this test for assessing BSA content in viral vaccines same as MMR vaccines. PMID- 18398307 TI - Protecting Indian schoolchildren against rabies: pre-exposure vaccination with purified chick embryo cell vaccine (PCECV) or purified verocell rabies vaccine (PVRV). AB - Although rabies can be effectively prevented by means of preexposure or post exposure prophylaxis, in India, an estimated 17,000 to 20,000 human rabies deaths occur annually. Tragically, 50% of these victims are children under the age of 15. In addition to immediate post-exposure prophylaxis measures, including active and passive immunization, pre-exposure vaccination using tissue culture vaccines is a safe and effective but highly underutilized method of preventing rabies in humans living or working in areas at risk. This study assessed the safety and immunogenicity of Purified Chick Embryo Cell Vaccine (PCECV) and Purified Verocell Rabies Vaccine (PVRV), administered as a three-dose intramuscular pre exposure regimen on days 0, 7 and 28 in 175 healthy schoolchildren. PCECV was administered after reconstitution using either 1.0 mL or 0.5 mL (half the diluent volume) and PVRV was given after reconstitution with 0.5 mL. Vaccine safety was assessed observer-blind, including pain assessment with a validated visual analogue scale for children. Rabies virus neutralizing antibody (RVNA) concentrations were measured on day 49 by RFFIT. All children developed adequate RVNA concentrations above 0.5 IU/mL. Solicited local and systemic reactions were within the range expected, pain after vaccination was reported in 2 to 12% of study subjects, fever was reported in 2 to 5%. There was no statistical difference by vaccination group or vaccination day. No unexpected or serious adverse event was reported during the study. In conclusion, PCECV and PVRV are safe and immunogenic when administered intramuscularly for pre-exposure prophylaxis of rabies in children. A 1.0 mL dilution volume for PCECV was as well tolerated as PVRV or PCECV reconstituted in half the volume. PMID- 18398308 TI - Immunogenicity of a two-component (PT & FHA) acellular pertussis vaccine in various combinations. AB - Immunogenicity data for the pertussis components of the French diphtheria-tetanus two component acellular pertussis vaccine (DTaP(2Fr)) obtained after primary series of immunizations were compiled from 75 study groups comprising 36 clinical trials or vaccination programs conducted between 1987 and 2006. DTaP(2Fr) vaccine was administered either as a standalone vaccine or as the backbone of several combination vaccines that included IPV, HepB and/or PRP-T antigens. Most of the variability in responses was associated with differences in the schedules, and to a lesser extent the geographical region where the study was performed, suggesting the importance of ethno-ecological factors. However the addition of other vaccine antigens did not affect the immunogenicity of the aP(2Fr) antigens. The immune responses to the PT and FHA antigens of the DTaP(2Fr) vaccine used in the Senegal efficacy trial, which established its good efficacy relative to a highly effective DTwP vaccine, was in the middle of the range of titers observed during other studies using the 2-4-6 months schedule conducted with the same vaccine. The consistent immunogenicity of the DTaP(2Fr) vaccine is accompanied by effectiveness in controlling pertussis disease in the areas where it is used on a large scale with good vaccination coverage. PMID- 18398309 TI - The multifaceted small RNAs. AB - Small RNA molecules are now being recognized as master regulators of gene activity. Although known initially for their gene-silencing functions, recent discoveries have revealed that small RNAs can also activate gene expression. These two opposing activities of gene silencing and activation together constitute an integrated gene regulatory mechanism. PMID- 18398310 TI - DNA methylation changes in schizophrenia and bipolar disorder. AB - The etiology of the major psychotic disorders, including schizophrenia and bipolar disorder, remains poorly understood. Postmortem brain studies have revealed altered expression of multiple mRNAs, affecting neurotransmission, metabolism, myelination and other functions. Epigenetic mechanisms could be involved, because for a limited number of genes, the alterations of mRNA levels have been linked to inverse DNA methylation changes at sites of the corresponding promoters. However, results from independent studies have been inconsistent, and when expressed in quantitative terms, disease-related methylation changes appear to be comparatively subtle. A recent study identified approximately 100 loci with altered CpG methylation in schizophrenia or bipolar disorder, the majority of which were gender-specific. Additional work will be necessary to clarify the origin and timing of these methylation changes in psychosis and to determine the specific cell types affected in the diseased brain. PMID- 18398311 TI - Epigenetic gene silencing in the Wnt pathway in breast cancer. AB - Breast cancer is one of the most common malignancies in women. Despite advances in treatment of endocrine-dependent tumors, the complete molecular basis of transformation is still unknown. What is clear is that a variety of genetic lesions and epigenetic modifications are present in the neoplasm. Disregulation of several signaling pathways is known to be associated with breast cancer development, among them is the wingless and integration site growth factor (Wnt) pathway. While genetic mutations of certain components of this pathway, such as APC, are significant contributing factors for colorectal cancers, they are typically not the predominate mechanism associated with breast cancer. Instead, it appears that DNA hypermethylation leads to aberrant regulation of the Wnt pathway in breast cancer, and as such, this review focuses on the epigenetic regulation of Wnt pathway components in breast cancer. PMID- 18398321 TI - Renin-angiotensin system, natriuretic peptides, obesity, metabolic syndrome, and hypertension: an integrated view in humans. AB - The obesity pandemic is closely related to hypertension and metabolic syndrome. Visceral adipose tissue plays a key role in the metabolic and cardiovascular complications of being overweight. The pathophysiological link between visceral adiposity and cardiometabolic complications focuses on insulin sensitivity, sympathetic nervous system, renin-angiotensin-aldosterone system (RAAS) and, only recently, on cardiac natriuretic peptide system (CNPS). RAAS and CNPS are endogenous antagonistic systems on sodium balance, cardiovascular system, and metabolism. The circulating RAAS is dysregulated in obese patients, and adipose tissue has a full local renin-angiotensin system that is active at local and systemic level. Adipocyte biology and metabolism are influenced by local renin angiotensin system, with angiotensin II acting as a 'growth factor' for adipocytes. CNPS induces natriuresis and diuresis, reduces blood pressure, and, moreover, has powerful lipolytic and lipomobilizing activity in humans but not in rodents. In obesity, lower plasmatic natriuretic peptides levels with increasing BMI, waist circumference, and metabolic syndrome have been documented. Thus, reduced CNPS effects coupled with increased RAAS activity have a central role in obesity and its deadly complications. We propose herein an integrated view of the dysregulation of these two antagonistic systems in human obesity complicated with hypertension, metabolic syndrome, and increased cardiovascular risk. PMID- 18398312 TI - WAMIDEX: a web atlas of murine genomic imprinting and differential expression. AB - The mouse is an established model organism for the study of genomic imprinting. Mice with genetic material originating from only one parent (e.g., mice with uniparental chromosomal duplications) or gene mutations leading to epigenetic deficiencies have proven to be particularly useful tools. In the process of our studies we have accumulated a large set of expression microarray measurements in samples derived from these types of mice. Here, we present the collation of these and third-party microarray data that are relevant to genomic imprinting into a Web Atlas of Murine genomic Imprinting and Differential EXpression (WAMIDEX: https://atlas.genetics.kcl.ac.uk). WAMIDEX integrates the most comprehensive literature-derived catalog of murine imprinted genes to date with a genome browser that makes the microarray data immediately accessible in annotation-rich genomic context. In addition, WAMIDEX exemplifies the use of the self-organizing map method for the discovery of novel imprinted genes from microarray data. The parent-of-origin-specific expression of imprinted genes is frequently limited to specific tissues or developmental stages, a fact that the atlas reflects in its design and data content. PMID- 18398322 TI - Aggressive blood pressure control and stroke prevention: role of calcium channel blockers. AB - Cerebrovascular disease is a major cause of morbidity and mortality worldwide and its prevalence is expected to increase as a result of projected demographic trends. Stroke is one of the leading causes of disability and death of over 30 million people each year worldwide. Hypertension is the most important modifiable risk factor for stroke. Recent data indicate that treatment with antihypertensive drugs reduces the incidence of all strokes in men (by 34%), women (by 38%), the elderly (by 36%), including those older than 80 years (by 34%), younger persons, those with systolic and diastolic hypertension, persons with isolated systolic hypertension, and those with a history of stroke or transient ischemic attack (by 28%). Furthermore, several large, prospective, randomized, clinical outcome trials have shown that calcium channel blockers (CCBs) are effective and safe antihypertensive drugs compared with placebo and reduce the cardiovascular morbidity and mortality of treated patients. Moreover, when CCBs were compared with conventional antihypertensive drugs they demonstrated similar blood pressure lowering effects and similar reductions in cardiovascular morbidity and mortality, with the exception of a higher incidence of heart failure and fatal myocardial infarction in some studies. Considering all the evidence available today, however, these drugs should be considered safe for the treatment of the uncomplicated hypertensive patient in combination with other drugs. They can also be used as first-line therapy for older, stroke-prone hypertensive patients. The aim of this review is to summarize the role of CCBs in the prevention of stroke. PMID- 18398323 TI - Neuroprotection by angiotensin receptor blockers? PMID- 18398324 TI - Pleiotropic effects of telmisartan: still more to come? PMID- 18398325 TI - Is cardiac hypertrophy a required compensatory mechanism in pressure-overloaded heart? PMID- 18398326 TI - Blood pressure and mortality among Chinese patients with cardiovascular disease. AB - OBJECTIVE: To examine the association between blood pressure (BP) and mortality among patients with a history of cardiovascular disease (CVD) in China. METHODS: We conducted a prospective cohort study among 4195 CVD patients aged 40 years and older. Data on BP and covariables were obtained at a baseline examination in 1991 and follow-up evaluation was conducted in 1999-2000 using standard protocols. RESULTS: After adjustment for important covariables, a significant and linear relationship was observed between BP level and mortality from CVD and all-causes. For example, compared with those with a systolic BP less than 120 mmHg, patients with a systolic BP of 120-129, 130-139, 140-159, 160-179, and at least 180 mmHg had relative risks (95% confidence interval) of 1.28 (0.92, 1.78), 1.62 (1.19, 2.20), 2.09 (1.58, 2.77), 2.31 (1.73, 3.10), and 2.66 (2.01, 3.53) for CVD mortality, and 1.08 (0.84, 1.38), 1.26 (1.00, 1.60), 1.44 (1.17, 1.79), 1.57 (1.25, 1.96), and 1.86 (1.50, 2.30) for all-cause mortality (both P values < 0.0001 for linear trends), respectively. The relationship between BP and mortality was slightly stronger for systolic BP compared with diastolic BP or pulse pressure. Lowering BP to a normal level in hypertensive patients could prevent 55.5% of CVD mortality and 31.2% of all-cause mortality among individuals with a history of CVD. CONCLUSION: These data indicate that there is a strong, independent, and positive association between BP and mortality among patients with a history of CVD. Furthermore, lowering of BP should be an important approach for preventing premature deaths in this population. PMID- 18398327 TI - Comparison of body mass index with waist circumference, waist-to-hip ratio, and waist-to-stature ratio as a predictor of hypertension incidence in Mauritius. AB - OBJECTIVE: Comparison of BMI with waist circumference, waist-to-hip ratio (WHR), and waist-to-stature ratio (WSR) as a predictor of hypertension incidence. METHODS: A total of 1658 men and 1976 women of Mauritian Indian and Mauritian Creole ethnicity, aged 25-74 years, free of hypertension, diabetes, cardiovascular disease, and gout at baseline in 1987 or 1992, were re-examined in 1992 and/or 1998 using the same survey methodology. Hazard ratios (HRs) for hypertension incidence were estimated applying an interval censored survival analysis (R program) using age as timescale based on baseline obesity indicators. RESULTS: A total of 787 incident hypertension cases were identified during the follow-up. HRs for hypertension incidence adjusting for baseline systolic blood pressure and cohort corresponding to a 1 SD increase in BMI, waist circumference, WHR, and WSR were 1.20 (1.24), 1.19 (1.21), 1.14 (1.10), and 1.20 (1.26) in Mauritian Indian men (women) and 1.23 (1.32), 1.34 (1.23), 1.41 (1.13), and 1.43 (1.33) in Mauritian Creoles, respectively, indicating that all obesity indicators significantly predicted hypertension incidence except for WHR in Mauritian Creole women. Paired homogeneity tests showed that there was no difference between BMI and the other three indicators for most of the comparisons with two exceptions: WSR was stronger than BMI (P = 0.002) in Mauritian Creole men but BMI was stronger than WHR (P = 0.047) in Mauritian Indian women in predicting the incident cases of hypertension. CONCLUSION: The relation of the development of hypertension with BMI was as strong as that with central obesity indicators in the population studied. PMID- 18398328 TI - Prevalence of microalbuminuria in the general population of Seychelles and strong association with diabetes and hypertension independent of renal markers. AB - OBJECTIVE: Few studies have examined microalbuminuria (MAU) in non-western populations. We assessed the prevalence of MAU in the general population of a middle-income country in the African region and relationships between MAU and selected cardiovascular risk factors. METHODS: An examination survey was conducted in a sample representative of the entire population aged 25-64 years in the Seychelles. MAU adjusted for urine creatinine concentration was measured on the second morning urine using a semiquantitative point-of-care analyzer. RESULTS: A total of 1255 persons attended the survey (participation rate of 80.2%). The age-adjusted prevalence of MAU was 11.4%. At age 25-64 years, the prevalence of MAU was 5% in persons without diabetes and hypertension, 20% in persons with either condition and 41% in persons with both conditions. The overall prevalence of stages 3-4 chronic kidney disease was low at 3.2%. In multivariate analysis, MAU was associated with age [odds ratio (OR) 1.24 for a 10 year increase; 95% confidence interval (CI): 1.02-1.52], hypertension stage I (2.0; 1.1-3.8) and stage II (4.5; 2.3-8.6), obesity (1.7; 1.0-2.8) and diabetes (3.0; 1.9-4.9). These associations were virtually unchanged upon further adjustment for markers of renal function such as serum creatinine, serum cystatin C and calculated renal function. CONCLUSION: The prevalence of MAU was high in this population, and MAU was strongly associated with several cardiovascular risk factors independently of renal function markers. These findings suggest that MAU could be a useful marker of cardiovascular risk in this population and help identify persons in need of a specific cardiovascular risk management. PMID- 18398329 TI - Pulse pressure or dipping pattern: which one is a better cardiovascular risk marker in resistant hypertension? AB - OBJECTIVE: Nocturnal blood pressure (BP) reduction and ambulatory pulse pressure (PP) are well known prognostic markers obtained from ambulatory BP monitoring (ABPM). The aim of this study is to investigate which one of these ABPM parameters is related to high cardiovascular risk profile in resistant hypertension, based on their associations with target organ damage (TOD). METHODS: Clinical-demographic, laboratory and ABPM variables were recorded in a cross-sectional study involving 907 resistant hypertensive patients. Nocturnal systolic BP reduction and 24-h PP were assessed both as continuous and dichotomized variables (PP at the upper tertile value: 63 mmHg). Statistical analyses included bivariate tests and multivariate logistic regression with each TOD as the dependent variable. RESULTS: Patients with the nondipping pattern and high 24-h PP shared some characteristics: they were older, had higher prevalence of cerebrovascular disease and nephropathy, higher office and 24-h BP levels, increased serum creatinine and microalbuminuria, and higher left ventricular mass index than their counterparts. Additionally, patients with high PP had a greater prevalence of diabetes and other TOD. In multivariate logistic regression, high PP was independently associated with all TODs even after adjustment for sex, age, BMI, cardiovascular risk factors, 24-h mean arterial pressure and antihypertensive treatment, whereas nondipping pattern was only associated with hypertensive nephropathy. Furthermore, PP was more strongly associated with the number of TOD than the nocturnal systolic blood pressure (SBP) fall. CONCLUSIONS: In a large group of resistant hypertensive patients, an increased 24-h PP shows a closer correlation with high cardiovascular risk profile than the nocturnal BP reduction. PMID- 18398330 TI - Masked hypertension in obstructive sleep apnea syndrome. AB - BACKGROUND: Ambulatory blood pressure (BP) monitoring (ABPM) detects subjects with normal clinic but high ambulatory 24-h BP, that is, masked hypertension. METHODS: One hundred and thirty newly diagnosed obstructive sleep apnea syndrome (OSAS) patients, free of recognized cardiovascular disease were included (111 men, age = 48 +/- 1 years, BMI = 27.6 +/- 0.4 kg/m, respiratory disturbance index (RDI = 42 +/- 2/h). Clinic BP, 24-h ABPM, baroreflex sensitivity (BRS), echocardiography and carotid intima-media thickness (IMT) were assessed. RESULTS: Forty-one patients (31.5%) were normotensive, 39 (30.0%) exhibited masked hypertension, four (3.1%) white-coat hypertension and 46 (35.4%) hypertension. Significant differences were found between normotensive, masked hypertensive and hypertensive patients in terms of BRS (10.5 +/- 0.8, 8.0 +/- 0.6 and 7.4 +/- 0.4 ms/mmHg, respectively, P < 0.001), carotid IMT (624 +/- 17, 650 +/- 20 and 705 +/ 23 microm, respectively, P = 0.04) and left ventricular mass index (37 +/- 1, 40 +/- 2 and 43 +/- 1 g/height2.7, respectively, P = 0.003). A clinic systolic BP more than 125 and a diastolic BP more than 83 mmHg led to a relative risk (RR) of 2.7 and a 90% positive predictive value for having masked hypertension. CONCLUSION: Masked hypertension is frequently underestimated in OSAS and is nearly always present when clinic BP is above 125/83 mmHg. PMID- 18398331 TI - Submegabase resolution of epistatically interacting quantitative trait loci for blood pressure applicable for essential hypertension. AB - OBJECTIVE: Although genetic mapping of quantitative trait loci for blood pressure to large chromosome segments is readily achievable, their final identification confronts formidable hurdles. Restriction of the genes lodging in one quantitative trait locus interval to experimental limitation can facilitate their positional cloning. We previously delineated several quantitative trait loci for blood pressure on chromosome 10 of Dahl salt-sensitive rats, but their chromosome delimitations were either large or not definitive. METHODS: In this study, we systematically and comprehensively constructed congenic strains with submegabase (Mb) genome resolution and analyzed their blood pressure by telemetry. RESULTS: Three quantitative trait loci have been conclusively delimited by three congenic strains, each independently lowering the blood pressure. Their intervals are demarcated by genomic regions between 350 and 910 kilobases (kb) in size. Two of the three quantitative trait loci share an epistatic relationship and are separated from one another by less than 170 kb. Two additional quantitative trait loci for blood pressure were also tentatively delineated and their intervals range from 520 kb to 1.75 Mb. Possible genes dwelling in each quantitative trait locus-interval number between 11 and 17. None of these genes is known to exert a functional impact on blood pressure. Work is underway to find candidate genes with mutations that could be responsible for the blood pressure effect. CONCLUSION: Novel diagnostic, prognostic, preventive and/or therapeutic targets for essential hypertension and hypertension-associated diseases are likely to emerge from the identification of these quantitative trait loci. Potential applications of these quantitative trait loci to humans are suggested from the positive results from several association studies, demonstrating the existence of quantitative trait loci in the broad homologous regions. PMID- 18398332 TI - Association of TNFRSF4 gene polymorphisms with essential hypertension. AB - BACKGROUND: Essential hypertension is a complex disorder that results from the interaction of a number of susceptibility genes and environmental factors. The TNFRSF4 (tumor necrosis factor receptor superfamily, member 4) gene was one of the genes that showed altered renal expression in long-term salt loading in mice. Moreover, association of the TNFRSF4 and TNFSF4 (tumor necrosis factor (ligand) superfamily, member 4) genes with myocardial infarction was recently reported. Since essential hypertension is a well-known risk factor for myocardial infarction, we hypothesized that TNFRSF4 could be a susceptibility gene for essential hypertension. METHODS: We performed a case-control study of TNFRSF4 in two independent population. RESULTS: Extensive investigation of single nucleotide polymorphisms of the entire gene suggested that it resided in one linkage disequilibrium block, and four single nucleotide polymorphisms in the 5' flanking region sufficiently represented major haplotypes. In the combined population, the frequency of the most frequent haplotype, C-C-A-A, was significantly lower (P = 8.07 x 10(-5)) and that of the second most frequent haplotype, C-T-G-A, was significantly higher (P = 6.07 x 10(-4)) in hypertensive subjects than in control subjects. This difference was observed only in female patients. The C-T-G-A haplotype showed a lower promoter activity than other haplotypes, suggesting a relationship with disease susceptibility. CONCLUSION: Our results suggest that TNFRSF4 is a female-specific susceptible gene for essential hypertension. PMID- 18398333 TI - Relationships among endogenous ouabain, alpha-adducin polymorphisms and renal sodium handling in primary hypertension. AB - OBJECTIVE: The basolateral Na pump drives renotubular reabsorption. In cultured renal cells, mutant adducins, as well as sub-nanomolar ouabain concentrations, stimulate the Na-K pump. METHODS: To determine whether these factors interact and affect Na handling and blood pressure (BP) in vivo, we studied 155 untreated hypertensive patients subdivided on the basis of their plasma endogenous ouabain or alpha-adducin genotype (ADD1 Gly460Trp-rs4961). RESULTS: Under basal conditions, proximal tubular reabsorption and plasma Na were higher in patients with mutated Trp ADD1 or increased endogenous ouabain (P = 0.002 and 0.05, respectively). BPs were higher in the high plasma endogenous ouabain group (P = 0.001). Following volume loading, the increment in BP (7.73 vs. 4.81 mmHg) and the slopes of the relationship between BP and Na excretion were greater [0.017 +/ 0.002 vs. 0.009 +/- 0.003 mmHg/(muEq min)] in ADD1 Trp vs. ADD1 Gly carriers (P < 0.05). BP changes were similar, whereas the slopes of the relationship between BP and Na excretion were lower [0.016 +/- 0.003 vs. 0.008 +/- 0.002 mmHg/(muEq min)] in patients with low vs. high endogenous ouabain (P < 0.05). In patients with high endogenous ouabain, volume loading increased the BP in the ADD1 Trp group but not in the Gly group (P < 0.05). Thus, patients with ADD1 Trp alleles are sensitive to salt and tubular Na reabsorption remains elevated after volume expansion. CONCLUSION: With saline loading, BP changes are similar in high and low endogenous ouabain patients, whereas tubular Na reabsorption increases in the high endogenous ouabain group. Saline loading unmasks differences in renal Na handling in patients with mutant adducin or high endogenous ouabain and exposes an interaction of endogenous ouabain and Trp alleles on BP. PMID- 18398334 TI - Liddle's syndrome caused by a novel missense mutation (P617L) of the epithelial sodium channel beta subunit. AB - OBJECTIVE: The aim of the study was to search for mutations of SCNN1B and SCNN1G in an Italian family with apparently dominant autosomal transmission of a clinical phenotype consistent with Liddle's syndrome. METHODS: Genetic analysis was performed in the proband, his relatives, and 100 control subjects. To determine the functional role of the mutation identified in the proband, we expressed the mutant or wild-type epithelial sodium channel in Xenopus laevis oocytes. RESULTS: A novel point mutation, causing an expected substitution of a leucine residue for the second proline residue of the conserved PY motif (PPP x Y) of the beta subunit was identified in the proband. The functional expression of the mutant epithelial sodium channel in X. laevis oocytes showed a three-fold increase in the amiloride-sensitive current as compared with that of the wild type channel. CONCLUSION: This newly identified mutation adds to other missense mutations of the PY motif of the beta subunit of the epithelial sodium channel, thus confirming its crucial role in the regulation of the epithelial sodium channel. To our knowledge, this is the first report of Liddle's syndrome in the Italian population, confirmed by genetic and functional analysis, with the identification of a gain-of-function mutation not previously reported. PMID- 18398335 TI - Newly developed renin and prorenin assays and the clinical evaluation of renin inhibitors. AB - BACKGROUND: The last decade has seen the introduction of renin inhibitors and new plasma renin and prorenin assays, which has led to a better understanding of the tissue renin-angiotensin system. AIM OF THE STUDY: To clarify the consequences of these developments for the methodology and interpretation of measurements of renin and prorenin. METHODS: The principles and application of the newly developed immunosorbent assays (ISAs) are surveyed and the results are compared with those of enzyme-kinetic assays (EKAs). RESULTS AND CONCLUSIONS: Angiotensin (Ang) II in cardiac, renal and adrenal tissue is known to originate mainly from locally produced Ang I. Experimental evidence and theoretical considerations show that a simple relation between Ang II receptor occupancy, in tissue micromilieu, and the circulating levels of Ang II or renin may not exist. This supports the clinicians' view that the plasma level of renin tells more about the mechanisms regulating its release into the circulation than about the Ang II-dependency of hypertension. ISAs are a welcome addition to clinical studies of renin inhibitors. By comparing the results of ISAs with those of EKAs, the inhibitor bound forms of renin and prorenin can be distinguished from the unbound forms. ISAs also provide important information on the molecular basis of prorenin activation. We propose a single kinetic model to incorporate the conformational changes of prorenin induced by cryo-activation and acid-activation, and by binding to renin inhibitors. It explains why renin ISAs can overestimate the rise of renin in response to these drugs, and shows how to deal with this artefact. PMID- 18398337 TI - Vascular stiffness is related to superoxide generation in the vessel wall. AB - OBJECTIVES: Oxidative stress causes endothelial dysfunction and plays a major role in the pathogenesis of cardiovascular disease. Increased vascular stiffness is an intermediate phenotype in the development of cardiovascular disease. We hypothesized that vascular stiffness is partially determined by oxidative stress. METHODS: We examined 163 participants out of whom 80 had coronary artery disease. Vascular stiffness was assessed by pulse wave analysis, pulse wave velocity and measurement of aortic compliance by cardiac MRI. Circulating markers of oxidative stress and vascular superoxide generation in saphenous vein were measured. RESULTS: After adjustment for age, sex, BMI, heart rate, blood pressure and lipids only carotid-femoral pulse wave velocity and aortic compliance were different between patients and control group. Aortic compliance was reduced (11.4 +/- 6.3 vs. 13.9 +/- 7.3 ml x 10(-3) per mmHg; P = 0.035) and vascular superoxide generation increased (1.01 +/- 0.45 vs. 0.76 +/- 0.44 nmol/mg per min; P = 0.035) in patients with coronary artery disease compared with those without. In a multiple stepwise regression analysis, aortic compliance was determined by age (P < 0.001) and vascular superoxide production (P = 0.033). CYBA C242T and NOS3 G894T polymorphisms had additive effects on vascular superoxide generation (P = 0.026) and xanthine oxidase activity was increased in patients with CAD (P = 0.043). Genetic factors (P = 0.033) and xanthine oxidase activity (P < 0.001) were also related to aortic compliance. CONCLUSION: By measuring vascular superoxide generation and aortic compliance using cardiac MRI, we demonstrated a functional relationship between oxidative stress and vascular stiffness. Patients identified with high levels of vascular stiffness are most likely to benefit from strategies to reduce vascular oxidative stress. PMID- 18398336 TI - Silencing regulator of G protein signaling-2 (RGS-2) increases angiotensin II signaling: insights into hypertension from findings in Bartter's/Gitelman's syndromes. AB - OBJECTIVE: Regulator of G-protein signaling (RGS)-2 is a regulator of angiotensin II (Ang II) signaling. In Bartter's syndrome/Gitelman's syndrome (BS/GS), we have demonstrated increased RGS-2 levels and blunted Ang II signaling which contribute to their reduced vasomotor tone and remodeling. The present study investigates the effect of silencing RGS-2 in fibroblasts from six BS/GS patients on intracellular Ca2+ (CaI2+) mobilization and extracellular signal-regulated kinase (ERK) 1/2 phosphorylation, established Ang II-mediated responses. METHODS: Fibroblasts were RGS-2 silenced by transfecting chemically synthesized small interfering RNA. Silencing efficiency and Ang II-induced ERK 1/2 phosphorylation were evaluated by western blot and Ang II-induced Cai2+ using Fura-2 AM. RESULTS: RGS-2 expression in not silenced BS/GS fibroblasts from patients is increased compared with healthy controls [0.34 +/- 0.02 vs. 0.19 +/- 0.01 densitometric units (d.u.), P = 0.0005]. Silencing RGS-2 in BS/GS patients was achieved to the level of controls. Ang II-induced Cai2+ release and ERK 1/2 phosphorylation were reduced in not silenced cells from BG/GS patients compared with controls (112.16 +/- 13.2 vs. 130.33 +/- 13.64 mmol/l, P = 0.011 and 0.64 +/- 0.08 vs. 0.91 +/- 0.03 mmol/l, P < 0.006, respectively). Silencing RGS-2 in BS/GS patients increased Ang II-induced Cai2+ release and ERK 1/2 phosphorylation in silenced cells compared with not silenced cells [59.3 +/- 10.8 (peak-basal) vs. 40.5 +/- 14.1 nmol/l, P = 0.017 and 0.84 +/- 0.06 vs. 0.64 +/- 0.08 nmol/l, P < 0.03, respectively], whereas they were not different compared with controls (60.1 +/- 4.3 and 0.91 +/- 0.03 nmol/l). Integrating the Cai2+ response over time showed increased Cai2+ area under the curve (AUC) of BS/GS silenced cells compared with that of not silenced cells (P = 0.013). CONCLUSION: This is the first report of silencing RGS-2 effect on Ang II signaling in a human clinical condition of altered vascular tone regulation and remodeling and establishes RGS-2 as a key regulatory element of Ang II signaling in humans. PMID- 18398338 TI - Reactive oxygen species mediate oxidized low-density lipoprotein-induced endothelin-1 gene expression via extracellular signal-regulated kinase in vascular endothelial cells. AB - BACKGROUND: Oxidized low-density lipoprotein (oxLDL) promotes expression and secretion of endothelin-1 (ET-1), however, the precise mechanism involved is unclear. This study was designed to identify the regulatory mechanism of oxLDL induced ET-1 expression in endothelial cells. METHODS: ET-1 mRNA expression, secretion and promoter activity were evaluated by reverse transcriptase-PCR (RT PCR), enzyme immunometric and luciferase assays, respectively. RESULTS: oxLDL (35 microg/ml) significantly enhanced reactive oxygen species (ROS), mRNA expression, secretion and promoter activity of ET-1 in human umbilical vein endothelial cells (HUVECs), all of which were nullified by the antioxidant N-acetyl cysteine (NAC). oxLDL stimulated the extracellular signal-regulated kinase (ERK) phosphorylation in HUVECs, which was blocked by NAC and the mitogen-activated protein/extracellular signal-regulated kinase (MEK) inhibitor PD98059. NAC and PD98059 stopped oxLDL-elicited increase in mRNA expression, secretion and promoter activity of ET-1. Fusion plasmids with decreasing length of 5'-flanking sequence of ET-1 from -566 bpLuc to -250 bpLuc displayed increased luciferase activity after 24 h of oxLDL treatment. Interestingly, fusion plasmid from -233 and -185 bpLuc significantly reduced the luciferase activity in control and oxLDL treated HUVECs. In addition, transfection of the reporter construct -250Luc, which contains a 2 bp mutation at activator protein-1 site, abolished both basal and oxLDL-stimulated ET-1 promoter activities. CONCLUSION: Collectively, our data favor the notion that oxLDL stimulates ERK phosphorylation via ROS accumulation, which in turn stimulates vascular endothelial transcriptional factor activator protein-1 and ET-1 expression as well as secretion. PMID- 18398339 TI - Effects of telmisartan, a unique angiotensin receptor blocker with selective peroxisome proliferator-activated receptor-gamma-modulating activity, on nitric oxide bioavailability and atherosclerotic change. AB - OBJECTIVE: Telmisartan is a unique angiotensin II (Ang II) receptor blocker (ARB) with selective peroxisome proliferator-activated receptor-gamma (PPAR gamma). We therefore investigated the effects of telmisartan on endothelial function and atherosclerotic change in genetically hyperlipidemic rabbits, compared with candesartan, an ARB without PPAR gamma activity. METHODS: A total of 30 Watanabe heritable hyperlipidemic (WHHL) rabbits equally derived (n = 6 each) were treated with (1) vehicle (control), (2) GW9662, a PPAR gamma antagonist (0.5 mg/kg per day), (3) telmisartan (5 mg/kg per day), (4) telmisartan + GW9662, (5) candesartan (5 mg/kg per day) for 8 weeks. After treatment, acetylcholine (ACh) induced nitric oxide production was measured as a surrogate for endothelium protective function, and vascular nitrotyrosine (a product of superoxide and nitric oxide) was measured for assessing dysfunctional endothelial nitric oxide synthase activity. Plaque area was quantified by histology. RESULTS: Telmisartan increased ACh-induced nitric oxide by 5.5 nmol/l, significantly more than control. Interestingly, cotreatment with GW9662 significantly attenuated telmisartan-induced ACh-induced nitric oxide almost to the levels observed with candesartan. Vascular nitrotyrosine concentration was 1.4 pmol/mg protein in the control group and significantly higher than that in the telmisartan or candesartan group. The lowest nitrotyrosine concentration was observed in the telmisartan group, which was significantly lower than that in the candesartan or telmisartan + GW9662 group. Histology of the thoracic aorta revealed that the plaque area was more significantly decreased in the telmisartan group than in the candesartan or telmisartan + GW9662 group. CONCLUSION: In addition to a class effect of ARBs, telmisartan may have additional effects on nitric oxide bioavailability and atherosclerotic change through its PPAR gamma-mediated effects in genetically hyperlipidemic rabbits. PMID- 18398340 TI - Inhibition of cardiovascular cell proliferation by angiotensin receptor blockers: are all molecules the same? AB - BACKGROUND: Few studies have directly compared the effects of different angiotensin receptor blockers (ARBs) on mechanisms involved in the pathogenesis of cardiovascular disease. METHODS: We studied the ability of different ARBs to inhibit the proliferation of vascular smooth muscle cells (VSMCs) and cardiac fibroblasts (CFs) in continuous culture and in quiescent cells stimulated to proliferate by platelet-derived growth factor (PDGF) and insulin. We also investigated whether the antiproliferative effects of ARBs depended on their ability to block angiotensin II receptors or activate the peroxisome proliferator activated receptor gamma (PPAR gamma). RESULTS: Dose-response studies showed that candesartan, eprosartan, and irbesartan had little or no effect on the proliferation of VSMC or CF in continuous culture even when tested at concentrations as high as 10 mumol/l or when tested in cells stimulated with PDGF/insulin. In contrast, telmisartan inhibited VSMC and CF proliferation by 50 70% (P < 0.05) in a dose-dependent and reversible fashion and significantly inhibited the increases in cyclin D1 levels and cell proliferation induced by PDGF/insulin. Antiproliferative effects of telmisartan were also observed in Chinese hamster ovary (CHO-K1) cells that lack functional angiotensin II receptors and in human VSMCs treated with the PPAR gamma antagonist GW9662. CONCLUSION: Telmisartan, but not candesartan, irbesartan, or eprosartan, can significantly inhibit the proliferation of VSMC and CF in culture when tested at concentrations near those that can be achieved in plasma with usual oral dosing. Telmisartan can also inhibit the proliferation of cells that lack angiotensin II receptors and cells treated with a PPAR gamma antagonist suggesting that the antiproliferative effects of telmisartan may involve more than just angiotensin II receptor blockade or activation of PPAR gamma. PMID- 18398341 TI - Plasma and urine aldosterone to plasma renin activity ratio in the diagnosis of primary aldosteronism. AB - OBJECTIVE: To establish the best cut-off value of the aldosterone (ALD)/plasma renin activity (PRA) ratio when screening patients for primary aldosteronism. One hundred and six patients with primary aldosteronism and 100 essential hypertensive patients were investigated in rigorous standardized conditions. METHODS: The ALD/PRA cut-off values were calculated from both the plasma and urine ALD/PRA ratio and analyzed by receiver operating characteristic (ROC) curve. In patients with PRA below 0.2 ng/ml/h [our radioimmunoassay detection limit], values were calculated both with PRA levels set at 0.2 ng/ml/h ('adjusted') and with PRA levels detected ('unadjusted') in the assay. RESULTS: ROC analysis on the ALD/PRA ratio indicated that the best performance was obtained when the plasma ALD (ng/dl)/PRA ratio was used in comparison with that observed in the urine ALD (microg/day)/PRA ratio. In patients with primary aldosteronism, as a whole group, the cut-off value of 69 corresponded to the best compromise value between sensitivity (96%) and specificity (85%), with and without PRA adjustment. In patients with aldosterone-producing adenoma, the cut off to obtain 100% sensitivity with high specificity (85%) proved to be 69, with and without PRA adjustment. In patients with bilateral adrenal hyperplasia, both with and without PRA adjustment, the best compromise between sensitivity (94%) and specificity (86%) was a cut-off value of 71. CONCLUSION: The best cut-off to identify patients with primary aldosteronism, corresponding to 69, was obtained by using the plasma ALD/PRA ratio. Adjustment of PRA to 0.2 ng/ml/h does not interfere with calculation of the plasma ALD/PRA ratio cut-off. PMID- 18398342 TI - Adrenal vein sampling for primary aldosteronism: the assessment of selectivity and lateralization of aldosterone excess baseline and after adrenocorticotropic hormone (ACTH) stimulation. AB - BACKGROUND: Adrenal vein sampling is crucial for identifying the primary aldosteronism subtypes, but the cutoff values for ascertaining selectivity of catheterization and lateralization of aldosterone secretion remain controversial. OBJECTIVES: To investigate the safety of adrenal vein sampling, the cutoff values for the selectivity and lateralization indexes, and the effect of adrenocorticotropic hormone stimulation on selectivity index and lateralization index performance. DESIGN: We assessed the proportion of selective adrenal vein sampling at different selectivity index cutoff values in 151 consecutive patients with primary aldosteronism undergoing bilaterally simultaneous adrenal vein sampling. Aldosterone-producing adenoma was diagnosed on the basis of the evidence of primary aldosteronism and lateralized aldosterone secretion, adenoma at pathological examination, and normokalemia, and correction of primary aldosteronism and cure or improvement of hypertension at follow-up. In 44 patients with bilaterally selective adrenal vein sampling and unequivocal diagnosis of aldosterone-producing adenoma on the basis of all these criteria, we examined the cutoff values of the lateralization index for assessing the lateralization of aldosterone excess and the effect of adrenocorticotropic hormone stimulation on selectivity index and lateralization index. RESULTS: Adrenal vein rupture occurred in one case (0.7%). Bilaterally selective adrenal vein sampling decreased steadily (from 79.9 to 40.2%) with increase in the selectivity index cutoffs from 1.1 to 5.0. Likewise, the proportion of correctly identified aldosterone-producing adenomas decreased (from 95.5 to 43.2%) with increase in lateralization index cutoffs from 1.125 to 5.0. Adrenocorticotropic hormone improved the assessment of selectivity but exerted a confounding effect on lateralization index. CONCLUSION: Adrenal vein sampling is safe; increasing the selectivity index cutoffs lowers the number of usable adrenal vein samplings; higher lateralization index cutoff values lead to missing a proportion of aldosterone-producing adenomas. The improved selectivity rate provided by adrenocorticotropic hormone stimulation should be weighed against the loss of correct lateralization. PMID- 18398343 TI - Brain and peripheral angiotensin II type 1 receptors mediate renal vasoconstrictor and blood pressure responses to angiotensin IV in the rat. AB - OBJECTIVES: Angiotensin (Ang) IV was reported to increase renal cortical blood flow (CBF) via putative angiotensin IV receptor (AT4) stimulation but reduce total renal blood flow (RBF) via angiotensin II type 1 (AT1) receptors. We investigated the effect of Ang IV on simultaneously measured mean arterial pressure (MAP), RBF, and CBF. The possible involvement of AT1 or AT4 receptors, the possible natriuretic effect, and responses to central administration were also explored. METHODS AND RESULTS: Intravenous injections of Ang IV dose dependently increased MAP and decreased CBF and RBF; these effects were abolished by AT1 receptor blockade. These reductions in CBF and RBF highly correlated as did renal vascular responses to Ang II and fenoldopam. Ang IV did not induce renal vasodilation even following AT1 receptor blockade. Intrarenal Ang IV infusion reduced CBF and RBF but had no natriuretic effect. Central Ang IV administration induced an AT1-mediated immediate increase in MAP and renal vascular resistance and a secondary increase in RBF. AT4 selective ligands, LVV hemorphin-7 and AT4-16 (intravenous, intrarenal or intracerebroventricular), had no effects on MAP, RBF or urinary sodium excretion. Additional in-vitro experiments indicated that the majority of the Ang IV-sensitive aminopeptidase activity in kidney membranes is attributed to aminopeptidase-N. CONCLUSION: Insulin-regulated aminopeptidase (IRAP)/AT4 receptors are involved in neither the regulation of RBF or CBF nor in the handling of renal sodium. Ang IV increases MAP and induces renal vasoconstriction via stimulation of brain and peripheral AT1 receptors and may be involved in the regulation of renal blood flow and blood pressure. PMID- 18398344 TI - c-Flip overexpression reduces cardiac hypertrophy in response to pressure overload. AB - OBJECTIVE: Activation of Fas signaling has been associated with the development of cardiomyocyte hypertrophy. In the present study, we investigated the effects of increased expression of c-Flip, a natural modulator of Fas receptor signaling, in a mouse model of cardiac growth response to pressure overload. METHODS: A transgenic mouse overexpressing c-Flip in the heart was generated in FVB/N strain. Echocardiographic, hemodynamic, histological and molecular analyses were carried out under basal conditions and after transverse aortic constriction (TAC) induced pressure overload. RESULTS: Overexpression of c-Flip in ventricular heart tissue was functionally silent under basal conditions affecting neither cardiac morphology nor basal cardiac function. Transgenic mice were then subjected to pressure overload by TAC procedure. Under such conditions, c-Flip transgenic mice showed normal left ventricular function with a significantly reduced left ventricular hypertrophy compared with wild-type mice and reduced induction of the cardiac "fetal" gene programme. Further, analysis of intracellular signaling pathways indicated that c-Flip overexpression reduced phosphorylation of both the glycogen synthase kinase 3beta (GSK3 beta) and Akt as compared with controls. Finally, the reduction of the TAC-induced hypertrophy was not accompanied by significant apoptosis increase. CONCLUSION: Altogether, these findings indicate c Flip as a key regulator of the cardiac response to ventricular pressure overload. PMID- 18398345 TI - Different role of wave reflection magnitude and timing on left ventricular mass reduction during antihypertensive treatment. AB - OBJECTIVE: Regression of left ventricular (LV) mass during antihypertensive treatment has been associated with reduction in aortic augmentation index, a composite measure of peripheral wave reflection. The aim of this study was to clarify which of the two reflection factors, that is magnitude or timing, plays the dominant role in this regression. METHODS: We evaluated the reflection magnitude (RM; the reflected-to-forward pressure wave amplitude ratio), the round trip travel time of the pressure wave (a parameter for reflection timing), and the aortic pulse wave velocity (PWV) with echocardiographic LV mass in 61 hypertensive patients before and after 1-year standard medical treatment. RESULTS: Antihypertensive therapy significantly (P < 0.01) decreased brachial and aortic blood pressures and aortic PWV, reduced LV mass, and increased travel time. Neither increase in travel time nor decrease in PWV, however, was related to the reduction in LV mass. By contrast, treatment-induced change in RM was significantly correlated with change in LV mass; the correlation was particularly close in patients with LV hypertrophy (r = 0.61, P < 0.001). Only a marginal correlation was observed between the changes in RM and travel time. The association between RM decrease and LV mass reduction was independent of age, sex, changes in travel time and blood pressure, and use of renin-angiotensin system inhibitors (beta = 0.41, P = 0.001). CONCLUSION: Decreased wave RM contributes to LV mass regression more strongly than, and independently of, delayed reflection timing. Peripheral muscular arteries (from which reflection arises) appear to be more important therapeutic targets in regressing LV mass than central elastic arteries. PMID- 18398346 TI - G-protein inwardly rectifying potassium channels are involved in the hypotensive effect of I1-imidazoline receptor selective ligands. AB - OBJECTIVE: The present study examined the role of G-protein inwardly rectifying potassium (GIRK) channels in the depressor responses elicited by intracisternal injections of imidazoline-like drugs in anesthetized rabbits. METHODS AND RESULTS: Intracisternal injections of the I1-imidazoline receptor (I1R) selective ligands LNP509 (30 microg/kg) and LNP640 (2 microg/kg) (subthreshold doses), and of the GIRK channel opener flupirtine (30 microg/kg) did not affect mean arterial blood pressure (MAP). LNP509 and LNP640, however, elicited substantial depressor responses in rabbits pretreated with flupirtine (-17 +/- 2 and -18 +/- 1 mmHg, respectively, P < 0.05). Injection of higher doses of LNP509 (200 microg/kg) or LNP640 (10 microg/kg) elicited substantial reductions in MAP (-45 +/- 3 and -39 +/- 2 mmHg, respectively, P < 0.05) in naive rabbits. The depressor responses elicited by the higher doses of LNP509 or LNP640 were markedly diminished by pretreatment with the GIRK channel blocker tertiapin-Q (10 microg/kg) (-23 +/- 3 and -26 +/- 2 mmHg, respectively, P < 0.05 compared with nonpretreated rabbits), whereas tertiapin-Q (10 microg/kg) did not affect MAP by itself. Maximal-specific binding (Bmax) of the I1R ligand [I]LNP911 to PC12 cell membranes (296 +/- 59 fmol/mg protein) was enhanced by flupirtine pretreatment whereas it was reduced by tertiapin-Q pretreatment (687 +/- 122 and 68 +/- 21 fmol/mg protein, respectively, P < 0.05 vs. control binding). CONCLUSION: These findings demonstrate that the modulation of GIRK channels affects I1R's function and raise the possibility that GIRK channels, and I1Rs are parts of a single proteic complex. PMID- 18398347 TI - The central hypotensive effect induced by alpha 2-adrenergic receptor stimulation is dependent on endothelial nitric oxide synthase. AB - OBJECTIVE: The aim of the present study was to determine whether the central antihypertensive effect of drugs that act via central alpha 2-adrenergic receptors is mediated by the nitric oxide-ergic system. METHODS: The hypotensive effects of dexmedetomidine, a 'pure' alpha2-adrenergic agonist, were compared in endothelial nitric oxide synthase knockout and wild-type control mice. RESULTS: When injected intravenously (5 mug/kg) in wild-type mice, dexmedetomidine elicited a depressor response (60 +/- 4 to 34 +/- 1 mmHg, P < 0.05), but had no hypotensive effect in endothelial nitric oxide synthase (eNOS) knockout mice (84 +/- 7 to 84 +/- 7 mmHg, P > 0.05). In the presence of N-omega-nitro-L-arginine, a nonselective nitric oxide synthase (NOS) blocker that does not cross the blood brain barrier, the hypotensive effect of dexmedetomidine was not abolished (Delta MAP = 21 +/- 2 mmHg vs. Delta MAP = 26 +/- 3 mmHg, P > 0.05). CONCLUSIONS: It is concluded that the central cardiovascular effects of alpha 2-adrenergic agonists, such as dexmedetomidine, require an intact expression of eNOS within the brain. This study raises the interesting question of whether central eNOS itself might be considered as a target for new cardiovascular drugs regardless of any activation of alpha 2-adrenergic receptors. PMID- 18398348 TI - Reduced arterial stiffness may contribute to angiotensin-converting enzyme inhibitor induced improvements in walking time in peripheral arterial disease patients. AB - OBJECTIVE: Claudication is a debilitating consequence of peripheral arterial disease. Evidence is accumulating to suggest that large artery stiffness may influence peripheral perfusion and walking time through effects on peripheral hemodynamics as well as microvascular structure and function. We have previously shown that the angiotensin-converting enzyme inhibitor ramipril increased systemic arterial compliance by 64%, and increased maximum walking time by over 200% in patients with peripheral arterial disease. In the current analysis in the same patient cohort, we hypothesized that this relationship may, in part, be causal. METHODS: Forty patients with peripheral arterial disease [66 +/- 4 years (mean +/- SD); n = 20 per group] were randomized to ramipril, 10 mg once daily, or placebo for 24 weeks in a double-blind study. Maximum walking time was recorded during a standard treadmill test. Indices of arterial stiffness were assessed globally by systemic arterial compliance and augmentation index and regionally via central pulse wave velocity. RESULTS: Ramipril increased maximum walking time by 243% and improved arterial stiffness parameters by between 17 and 64% (all P < 0.001 compared with placebo). There were moderately strong correlations between the pre/post intervention change in maximum walking time and the change in indices of arterial stiffness (systemic arterial compliance, r = 0.65, P < 0.001; central pulse wave velocity, r = -0.57, P < 0.001; augmentation index, r = -0.79, P < 0.001; time to pressure augmentation, r = 0.52, P = 0.001). CONCLUSION: The present data support the hypothesis that the beneficial effects of ramipril on maximum walking time observed in our peripheral arterial disease population are, at least partly, a consequence of reduced arterial stiffness. PMID- 18398349 TI - Rule of thumb to calculate mean arterial pressure at the brachial artery level. PMID- 18398350 TI - Prevalence of hypertension and cardiovascular risk factors in Belgian civil employees: results of the screening during World Hypertension Day 2007. PMID- 18398351 TI - Teletransmission of home blood pressure monitoring: making it easier. PMID- 18398353 TI - Presbyopia-correcting intraocular lenses: what retinal surgeons should know. AB - BACKGROUND: Presbyopia-correcting intraocular lenses (IOLs) are becoming more popular among patients undergoing cataract surgery. Performing vitrectomy with these lenses can be challenging. METHODS: A review of the currently available presbyopia-correcting IOLs is performed. The problems with vitrectomy and presbyopia-correcting IOLs are identified, and suggestions are given on minimizing intraoperative complications. RESULTS AND CONCLUSIONS: As these lenses gain popularity among patients with cataracts, retinal surgeons need to be aware of the challenges of working through them when performing vitrectomy. However, with careful preoperative evaluation and planning, complications can be avoided. PMID- 18398355 TI - Early postoperative hypotony after 25-gauge sutureless vitrectomy with straight incisions. AB - PURPOSE: To evaluate the rate, duration, and associated complications of postoperative hypotony after 25-gauge sutureless vitrectomy. METHODS: In a retrospective, interventional, institutional case series, 25-gauge sutureless vitrectomy with straight incisions was performed on 111 eyes. Postoperative intraocular pressure (IOP) was measured routinely at 2 hours, 1 day, 1 week, and 1 month. Postoperative IOP, hypotony (IOP, <8 mmHg) rates, and complications associated with low IOP were assessed to determine outcome. RESULTS: Mean follow up +/- SD was 5.45 +/- 3.44 months. Mean IOP decreased significantly by 2 hours and on postoperative day 1 (P < 0.001) and increased to preoperative levels at week 1 (P = 0.48). Postoperative hypotony rates were 26.12%, 17.11%, and 8.10% at 2 hours, 1 day, and 1 week, respectively. The ratio of postoperative vitreous hemorrhage in eyes with and without hypotony on day 1 was 2:6 (P = 0.412). Two eyes with macular holes (MHs) filled inadequately with gas endotamponade that remained hypotonic for the first postoperative week underwent reoperation due to unsealed MHs. CONCLUSIONS: Twenty-five-gauge sutureless vitrectomy with straight incisions resulted in a hypotony rate of 26.12% at 2 hours that decreased to 17.11% on day 1 and to 8.10% at week 1 but did not increase the incidence of postoperative complications. PMID- 18398354 TI - Discrepancy between fluorescein angiography and optical coherence tomography in detection of macular disease. AB - PURPOSE: To compare high-resolution optical coherence tomography (OCT) and fluorescein angiography (FA) in detection of macular edema (ME) of various etiologies. METHODS: In a retrospective study over a 12-month period at one retina center, data for consecutive eyes that had undergone simultaneous conventional FA (HRA; Heidelberg Engineering, Vista, CA) and StratusOCT (Carl Zeiss Meditec, Dublin, CA) to rule out ME were reviewed. A subset of patients underwent additional examination with extremely high-resolution (6 microm)/ultrahigh-speed spectral OCT/scanning laser ophthalmoscopy (OTI, Inc., Toronto, Ontario, Canada). RESULTS: Of 1,272 eyes, 1,208 (94.97%) had the finding of ME or subretinal fluid confirmed by both techniques. There were 49 eyes (3.86%) for which FA showed dye leakage in the macular area and OCT showed normal foveal contour. Of 10 eyes in this group that underwent imaging with ultrahigh speed spectral OCT/scanning laser ophthalmoscopy, 8 had subtle diffuse lucencies in the retina. For 15 eyes (1.17%), OCT showed intraretinal and subretinal fluid, which was missed by FA. CONCLUSIONS: Both FA and high-resolution OCT are highly sensitive techniques and correlate well in detection of ME. However, there is a small chance that when performed alone they might miss existing subtle ME. PMID- 18398356 TI - Ocular surface fluid contamination of sutureless 25-gauge vitrectomy incisions. AB - PURPOSE: To determine whether 25-gauge transconjunctival sutureless vitrectomy incisions are more likely to allow the inflow of ocular surface fluid than sutured vitrectomy incisions. METHODS: Four groups of cadaver eyes underwent three-port vitrectomy in a laboratory setup. Groups A and B had 25-gauge vitrectomy, with and without conjunctival displacement, respectively, and were left unsutured. Group C had 25-gauge vitrectomy with suturing of the sclerotomies. Group D had 20-gauge sutured vitrectomy. In all groups, India ink was applied to the ocular surface after the procedure, and the intraocular pressure (IOP) was varied to simulate postoperative IOP changes. Histology of the incisions was performed. RESULTS: Ink particles were identified in the wound in over two-thirds of eyes that underwent 25-gauge sutureless vitrectomy. Conjunctival displacement had no effect on ink ingress. Sutured 25- and 20-gauge incisions did not demonstrate ink particles in the wound. CONCLUSIONS: Twenty five-gauge transconjunctival sutureless vitrectomy incisions, with or without conjunctival displacement, allowed the entry of ocular surface fluid into the eye in this laboratory model. Based on the limited number of eyes examined in this study, sutureless vitrectomy techniques may carry an increased risk of bacterial contamination that could predispose to endophthalmitis. Suturing may protect against ocular surface fluid ingress. PMID- 18398357 TI - Retinal function after vitrectomy. AB - PURPOSE: To study retinal function after vitrectomy. METHODS: Core vitrectomy was performed in 12 rabbits under standardized conditions using a vitreous cutting rate of either 600 or 1200 cuts/min. Full-field electroretinography (ERG) and multifocal electroretinography (mfERG) were performed pre- and postoperatively. Morphologic change was monitored by immunohistochemistry directed against glial fibrillary acidic protein (GFAP). RESULTS: Three days postoperatively, the b-wave amplitudes of all cone and rod responses of the ERG were significantly reduced in all vitrectomized eyes. At 28 days, the rod response was still reduced, but returned to normal by 58 days. No correlation was found between vitreous cutting speed and ERG findings. No reduction in the central cone function was detected in the mfERG. GFAP upregulation was found in the entire retina of vitrectomized eyes 3 days after surgery. GFAP expression was present after 28 and 58 days in eyes in which the vitreous cutting rate had been set to 600 cuts/min, but not in the 1200 cuts/min eyes. CONCLUSION: Pars plana vitrectomy transiently affects retinal function in rabbit eyes. Vitreous cutting speed is not related to the reduced function but appears inversely correlated to Muller cell activation, indicating that high-speed vitreous cutters are more lenient to the retina. PMID- 18398358 TI - Incidence of acute onset endophthalmitis following intravitreal bevacizumab (Avastin) injection. AB - PURPOSE: To report the incidence of acute endophthalmitis as a complication of intravitreal bevacizumab (Avastin) (IVB) injection in a tertiary vitreoretinal group practice. METHODS: A retrospective chart review of 5,233 consecutive eyes that underwent IVB injection at Retina Consultants of Alabama (RCA) from October 1, 2005, to August 31, 2007, was performed to identify cases of acute endophthalmitis. RESULTS: During the 23-month study interval, the overall incidence rate of postinjection endophthalmitis was 0.019% (1/5,233). In the single case of acute endophthalmitis, bacterial cultures revealed coagulase negative Staphylococcus (CNS) species. Visual acuity after treatment for endophthalmitis was improved (baseline 4/400) to 20/400 at two months after the initial IVB injection. CONCLUSIONS: Acute endophthalmitis is a rare potential complication of IVB. Prophylaxis with topical povidone-iodine and adherence to aseptic technique minimizes the risk of postinjection infection. SUMMARY STATEMENT: A retrospective review of 5,233 consecutive intravitreal injections of bevacizumab (Avastin) revealed only a single case of acute endophthalmitis. Adherence to aseptic technique and the use of povidone-iodine prophylaxis minimizes the risk of postinjection intraocular infection. PMID- 18398359 TI - Three-year anatomic and visual outcomes after vitrectomy for stage 4B retinopathy of prematurity. AB - PURPOSE: To assess anatomic and visual outcome following vitrectomy for stage 4B retinopathy of prematurity (ROP) after 3 years follow-up. METHODS: Retrospective analysis of 56 eyes (46 infants) with stage 4B ROP-related retinal detachment. Eyes underwent either lens-sparing vitrectomy (LSV) or combined lensectomy and vitrectomy (LV). Visual outcome after a minimum follow-up of 36 months was ascertained by using either Teller or Snellen acuities, or sweep visual evoked potential. RESULTS: LSV was performed in 42.9% and LV in 57.1% of eyes. Retinal reattachment was achieved in 73.2% overall (LV 71.8% versus LSV 75%) (P = 0.96). Ambulatory vision (VA better than 20/1900) and near reading vision (better than 20/800) were attained in 97.4% and 42.8% of eyes respectively. Attached retina, LSV, and pretreatment with retinal ablation were associated with a higher incidence of near reading or better vision compared to detached (P < 0.001), undergoing LV (P < 0.001), and non pretreated eyes (P = 0.011). CONCLUSION: After 3 years, more than 40% of eyes operated for ROP stage 4B in this series had a visual acuity compatible for near reading or better. Eyes undergoing LSV and with prior peripheral retinal ablation had better visual outcomes. PMID- 18398360 TI - Secondary ocular hypertension after intravitreal injection of 4 mg of triamcinolone acetonide: incidence and risk factors. AB - PURPOSE: To analyze the incidence of secondary ocular hypertension (SOH) after intravitreal triamcinolone acetonide (IVTA) injection and its risk predictors. METHODS: Retrospective review of charts for 219 consecutive patients receiving a 4-mg IVTA injection. RESULTS: One hundred fifty eyes of 150 patients who were followed for at least 3 months and met inclusion criteria were considered. Main indications for IVTA injection were neovascular age-related macular degeneration (79 eyes [52.7%]), choroidal neovascularization due to other etiologies (22 eyes [14.7%]), diabetic macular edema (14 eyes [9.3%]), central retinal vein occlusion (12 eyes [8.0%]), and branch retinal vein occlusion (8 eyes [5.3%]). SOH defined as intraocular pressure (IOP) of >or=21 mmHg was recorded for 32.0% of injected eyes at some point during a mean follow-up of 7.7 months. There was no association between SOH and age, sex, arterial hypertension, diabetes mellitus, indication for IVTA injection, prior cataract surgery, or concurrent photodynamic therapy. Although previous pars plana vitrectomy did not influence risk, peak IOP was lower in vitrectomized eyes (P = 0.044). Prior diagnosis of glaucoma was a significant risk factor for SOH (relative risk = 2.17; P = 0.004). In nonglaucomatous eyes, baseline IOP of >or=16 mmHg was associated with a higher risk of SOH (relative risk = 2.31; P = 0.003). Baseline IOPs of <12 mmHg, 12-14 mmHg, 15-17 mmHg, 18-20 mmHg, and >20 mmHg were associated with incidences of SOH of 11.1%, 25.4%, 40.0%, 46.2%, and 50.0% (P = 0.01), respectively. CONCLUSIONS: A 4-mg IVTA injection was associated with SOH in 32.0% of treated eyes. The risk of SOH was higher in eyes with previous glaucoma and higher baseline IOP. Peak IOP after IVTA injection was lower in vitrectomized eyes. Risk factor analysis may permit better individualization of the risk-benefit ratio for IVTA injection. PMID- 18398361 TI - Central retinal vein occlusion associated with cilioretinal artery occlusion. AB - PURPOSE: To describe the clinical characteristics and pathogenesis of central retinal vein occlusion (CRVO) associated with cilioretinal artery occlusion (CLRAO). METHODS: The study included 38 patients (38 eyes) who had CRVO associated with CLRAO and were seen in our clinic from 1974 to 1999. At their first visit to our clinic, all patients provided a detailed ophthalmic and medical history and underwent comprehensive ophthalmic evaluation, color fundus photography, and fluorescein fundus angiography. At each follow-up visit, the same ophthalmic evaluations were performed, except for fluorescein fundus angiography. RESULTS: Of 38 eyes, 30 had nonischemic CRVO, 5 had ischemic CRVO, and 3 had nonischemic hemi-CRVO. Patients with nonischemic CRVO were significantly younger (mean age +/- SD: 45.3 +/- 16.0 years) than those with ischemic CRVO (72.3 +/- 9.2 years; P = 0.001) and those with nonischemic hemi CRVO (64.7 +/- 7.5 years; P = 0.018). At least one third of the patients gave a definite history of episode(s) of transient visual blurring before the onset of constant blurred vision. Initially, the ophthalmoscopic and fluorescein angiographic findings were similar to those seen in CRVO and hemi-CRVO, except that all these eyes had retinal infarct in the distribution of the cilioretinal artery; its size and site varied widely. Fluorescein angiography typically showed only transient hemodynamic block and not the typical CLRAO. During follow-up, visual acuity improved markedly in nonischemic CRVO (P < 0.001) and nonischemic hemi-CRVO but deteriorated in ischemic CRVO. Retinopathy resolved spontaneously in 22 eyes with nonischemic CRVO (mean duration +/- SD: 42.0 +/- 101.0 months), in 2 eyes with ischemic CRVO (15.4 +/- 4.5 months), and in 1 eye with nonischemic hemi-CRVO. Retinociliary collaterals developed in 30% of eyes with nonischemic CRVO, in 40% of eyes with ischemic CRVO, and in 66% of eyes with nonischemic hemi CRVO. CONCLUSION: CRVO associated with CLRAO constitutes a distinct clinical entity. The pathogenesis of CLRAO in CRVO is due to transient hemodynamic blockage of the cilioretinal artery caused by a sudden sharp rise in intraluminal pressure in the retinal capillary bed (due to CRVO) above the level of that in the cilioretinal artery. PMID- 18398362 TI - Safety of intravitreal ketorolac and diclofenac: an electroretinographic and histopathologic study. AB - OBJECTIVE: To determine the clinical, histologic, and electroretinographic effects in the rabbit retina of escalating doses of two intravitreally delivered nonsteroidal anti-inflammatory drugs (NSAIDs): ketorolac and diclofenac. METHODS: Right eyes received a single 0.1 mL injection of either ketorolac (500-6000 microg/0.1 mL) or diclofenac (300-1500 microg/0.1 mL) prepared in balanced salt solution (BSS). Left eyes served as controls and received BSS. Dark- and light adapted electroretinograms (ERG) were obtained at baseline and 4 and 8 weeks postinjection. Enucleated eyes were examined histologically. RESULTS: Ophthalmic examinations demonstrated no signs of intraocular inflammation or retinal toxicity. Intraocular pressure measurements remained similar between NSAID injected and control eyes. Histologic and ERG studies of eyes injected with 6000 microg ketorolac and >or=500 microg diclofenac demonstrated toxicity. In contrast, doses up to 3000 microg ketorolac demonstrated enhanced b-wave amplitude responses. Delayed drug toxicity was observed for the highest doses of both NSAIDs. CONCLUSIONS: Intravitreal 3000 microg ketorolac and 300 microg diclofenac were nontoxic in this animal study, and may offer an effective and safer alternative to intravitreal corticosteroids. PMID- 18398363 TI - Central serous chorioretinopathy associated with sildenafil. AB - PURPOSE: : To investigate a suspected relationship between central serous chorioretinopathy (CSC) and sildenafil therapy. METHODS: : For this retrospective, observational case series, the authors reviewed over 1,500 case reports of sildenafil-associated ocular side effects from the postmarketing surveillance databases of the Food and Drug Administration (FDA), the World Health Organization (WHO), and the National Registry of Drug-Induced Ocular Side Effects. They also performed a Medline literature search for "retinopathy, retinal edema, macular edema" (terms associated with CSC) and "sildenafil." They identified 11 cases of CSC in men taking sildenafil. RESULTS: : In 8 of 11 cases, patients stopped sildenafil therapy when CSC occurred. In 6 of these 8 cases, patients' vision improved after sildenafil cessation. In 3 cases, effects recurred when patients resumed sildenafil therapy, but stopping therapy was not associated with improvement of CSC in every case. Two patients continued to experience CSC after sildenafil cessation. CONCLUSION: : Practitioners who see patients with refractory CSC should consider recommending cessation of sildenafil therapy; however, because of the cyclic nature of CSC, a causal relationship has not yet been established. Additional possible case reports can be sent to the National Registry of Drug-Induced Ocular Side Effects, the FDA, the WHO, or the manufacturer. PMID- 18398364 TI - Macular hole formation in patients with retinitis pigmentosa and prognosis of pars plana vitrectomy. AB - PURPOSE: To investigate macular hole (MH) formation in patients with retinitis pigmentosa (RP) and to determine surgical prognosis for these patients. METHODS: Data for four RP patients with MHs were retrospectively reviewed. Comprehensive ocular examinations, including electroretinography, Goldmann perimetry, color funduscopy, B-scan ultrasonic tomography, and optical coherence tomography (OCT), were performed. All patients underwent par plana vitrectomy followed by peeling of the posterior inner limiting membrane and gas tamponade. Preoperative best corrected visual acuity ranged from 20/1000 to 20/100. RESULTS: Typical full thickness MHs were evident by OCT for three patients. Two of these patients had posterior vitreous detachment or vitreoretinal traction; one had cystoid macular edema in the fellow eye. A fourth patient had extensive retinal detachment; ophthalmoscopy revealed a hole in the macular region. After surgery, significant improvement of visual acuity was recorded for three patients; the patient with retinal detachment had no change in visual acuity in the affected eye. OCT showed adequate sealing of the MHs. CONCLUSIONS: We report results of a clinical investigation including four RP patients with concomitant MHs. Vitreoretinal surgery is necessary for effective management of MHs in RP. PMID- 18398365 TI - Large-spot subthreshold infrared laser to treat diabetic macular edema. AB - PURPOSE: To evaluate the efficacy of a large-spot subthreshold infrared laser protocol to treat diabetic maculopathy. METHODS: In a prospective, fellow eye, controlled case series, all patients had clinically significant diabetic macular edema (DME) treated with a single application of subthreshold infrared (810 nm) laser. If bilateral disease was present, the fellow eye was treated with conventional macular laser. The study was to include 20 patients. Visual acuity and central macular thickness (CMT) measured by optical coherence tomography (OCT) were assessed in the study and fellow eyes at baseline and 6 months, and any changes were compared. RESULTS: The 11th patient developed a choroidal infarct with subsequent profound loss of vision immediately after treatment. The study was terminated prematurely at this point. For the remaining 10 patients, there was a trend toward improvement in visual acuity in the study eye compared with the fellow eye at the 6-month follow-up (median change: +1.5 letters for study eye vs -6.5 letters for fellow eye; P = 0.08). There was also significant improvement in OCT-measured CMT in the study eye (mean decrease, 117 microm) compared with deterioration in OCT-measured CMT in the fellow eye (mean increase, 24 microm; P = 0.02). CONCLUSION: This subthreshold infrared laser protocol led to improvement in OCT-measured CMT and stabilization of vision in most subjects. The current protocol is however unpredictable and should not be used in the treatment of DME without further modification. PMID- 18398366 TI - A masking device and method to create custom laser exposure patterns for verteporfin photodynamic therapy. AB - PURPOSE: To demonstrate a method for creating individualized laser exposure patterns for verteporfin photodynamic therapy (PDT) for peripapillary, juxtafoveal, and irregularly shaped lesions. METHODS: Laser beams from commercially available PDT lasers (Lumenis Opal [Lumenis, Inc., Yokneam, Israel (formerly Coherent Medical Group, Palo Alto, CA)] and Zeiss Visulas 690s [Carl Zeiss Jena GmbH, Jena, Germany] were masked using different patterns and household materials. The masking was tested for percentage of light transmission, fluence rate across the masked treatment area, and intensity drop-off at the edge of the mask. RESULTS: Of the materials tested, electrical tape was completely opaque, black card stock allowed 0.6% measured transmission, and yellow paper allowed 18% transmission of laser light. For the Lumenis Opal laser, the ratio of mask size to image size at the slit-lamp focal plane was 1:1. For the Zeiss Visulas laser, the ratio was 2:1; however, the exact ratio of the mask size to the image size was dependent on the spot size setting used. Unmasked areas of laser light had the expected fluence rate (i.e., no masking effect). The intensity drop-off at the mask edge was equivalent to the edge of the unmasked beam. CONCLUSION: Masks that individualize laser exposure patterns during PDT would allow safe treatment of irregular lesions and limit exposure of healthy tissue to risks of treatment. PMID- 18398367 TI - Human idiopathic epiretinal membranes express NGF and NGF receptors. AB - PURPOSE: Glial cells and fibroblasts (FBs) play a key role in epiretinal membrane (ERM) development and progression. Myofibroblasts (myoFBs), arising from these cells, can lead to the hypertrophic scars and tissue contraction observed in ERMs. Nerve growth factor (NGF) and transforming growth factor-beta1 (TGF-beta1) play a crucial role in FB activities. Therefore, the authors evaluated myoFBs in ERMs and NGF, trkA(NGFR and p75(NTR) expression, as well as TGF-beta1/TGF-betaRII levels in both ERMs and vitreous. METHODS: Eight idiopathic ERMs and vitreous were obtained from patients at the time of vitrectomy for macular pucker. Ten control vitreous were from donors. Biochemical and molecular analyses were performed to identify alpha-smooth muscle actin (alpha-SMA, a defined myoFB marker), NGF, trkA(NGFR)/p75(NTR), and TGF-beta1/TGF-betaRII. RESULTS: Every idiopathic ERM displayed alpha-SMA positive myoFBs, expressing NGF, trkA(NGFR), and p75(NTR). ERM vitreous showed a significant decrease in NGF protein coupled with a TGF-beta1 increase. In addition, vitreous cells showed an increase in trkA(NGFR)/p75(NTR) mRNA associated with a decrease in TGF-betaRII mRNA. CONCLUSIONS: Idiopathic ERMs were characterized by myoFBs. The expression of NGF, trkA, and p75 in local myoFBs associated with changes in ERM vitreous NGF suggests an involvement of NGF, as previously reported for TGF-beta1, in the evolution and myoFB-mediated contractile activity of ERMs. PMID- 18398368 TI - Neuroprotective effects of oral lamotrigine administration on rabbit retinas after pars plana vitrectomy and silicone oil injection. AB - PURPOSE: To investigate potential retinal neuroprotective effects of oral lamotrigine in rabbits after pars plana vitrectomy (PPV) and intravitreal silicone oil injection (SOI). METHODS: Twelve New Zealand rabbits (weight, 2.0 2.5 kg) underwent PPV with SOI on the right eye. For 30 days postoperatively, 6 rabbits received a daily oral dose of lamotrigine (25 mg/kg), and 6 rabbits received a daily oral dose of water. The animals were killed 30 days after surgery. All retinas were processed histologically, immunostained using glial fibrillary acidic protein (GFAP), and analyzed by fluorescence microscopy. Retina sections from all groups were analyzed by TUNEL for the presence of apoptosis and stained with hematoxylin-eosin for morphologic analysis and retina cell density measurements in each layer using a Zeiss Axiophot microscope and KS 400 software. RESULTS: Retinas from water-operated eyes showed a significant decrease in cell density associated with cell death compared with retinas from water-control eyes; cell density was reduced by 56% in the outer nuclear layer (ONL), 49% in the inner nuclear layer (INL), and 64% in the ganglion cell layer (GCL). Lamotrigine operated retinas showed a reduction in cell death when compared with water operated retinas; cell death was reduced by 52% in the ONL, 25% in the INL, and 56% in the GCL. Water-operated retinas showed TUNEL-positive cells and GFAP immunofluorescence throughout Muller cell processes; lamotrigine-operated retinas showed no TUNEL-positive cells and decreased GFAP staining when compared with water-operated retinas. CONCLUSIONS: PPV with SOI was associated with apoptosis of retinal cells and activation of glial cells in rabbit eyes. Oral lamotrigine administration provided protection against these effects. PMID- 18398369 TI - Inflammatory markers in nondiabetic and diabetic rat retinas exposed to ischemia followed by reperfusion. AB - PURPOSE: To examine the retinal inflammatory response to ischemia-reperfusion in nondiabetic and diabetic rats injected with either an omega-3-polyunsaturated fatty acid (docosahexaenoic acid [DHA]) or a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (pravastatin). METHODS: Diabetes was induced by an intraperitoneal injection of streptozocin, and retinal ischemia was induced by ligation of the optic nerve and vessels, followed by reperfusion for 1 hour or 24 hours. Five minutes before surgery, an intravenous injection of DHA, pravastatin, or vehicle (ethanol) was administered. The mRNA expressions of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, caspase-1, IL-1beta, P-selectin, vascular cellular adhesion molecule (VCAM)-1, and intercellular adhesion molecule (ICAM)-1 were compared between ischemic and nonischemic retinas as well as diabetic and nondiabetic nonischemic retinas. RESULTS: Ischemia induced increased expressions of TNF-alpha (P 5 cm after dose 5. These reactions were well tolerated and support revaccination. PMID- 18398384 TI - Frequency and risk factors for deep focus of infection in children with Staphylococcus aureus bacteremia. AB - BACKGROUND: Staphylococcus aureus bacteremia (SAB) in children may be associated with development of deep-seated foci of infection, often prompting extensive diagnostic testing. The objective of this study was to establish the frequency and risk factors for deep foci of infection from SAB in pediatric patients. METHODS: Medical charts of all children admitted with SAB to a tertiary-care center from January 1992 to June 2006 were reviewed. Study outcome was the presence of a deep focus of infection as documented by positive echocardiogram, bone imaging or abdominal imaging. RESULTS: We studied 298 children, of whom 190 (64%) had echocardiograms, 116 (39%) had abdominal imaging, and 103 (35%) had bone imaging. Forty-seven subjects (16%) had symptoms of a deep focus of infection on discovery of SAB, which then was confirmed by 1 of the 3 tests. Eleven (3.7%) additional subjects had a clinically unsuspected deep focus identified before discharge. All children with an unsuspected deep focus of infection had either an underlying medical condition that potentially obscured the diagnosis or a central venous catheter. More than 1 day of positive blood cultures was associated with an unsuspected deep-seated infection (P < 0.01). Endocarditis was uncommon (2.7%), and occurred only in children with known congenital heart disease or with a central catheter. CONCLUSIONS: Deep-seated infections from SAB in children are most often clinically apparent at discovery of bacteremia. Unsuspected deep-seated infection is uncommon and confined to specific hosts. Routine diagnostic imaging is not indicated in all children with SAB. PMID- 18398386 TI - Polymicrobial bloodstream infection in pediatric patients: risk factors, microbiology, and antimicrobial management. AB - BACKGROUND: Few studies focus on polymicrobial bloodstream infections (PBSIs) in children. In previous reports, children with PBSI frequently had complex underlying medical conditions and a high incidence of specific microorganisms, but systematic evaluation with controls was not performed. We postulated that specific clinical risk factors are associated with an increased risk of PBSI, and that illness may be more severe with these infections. Additionally, we suspected that routine empiric antimicrobial therapy may frequently be inadequate to treat the variety of pathogens in PBSI. METHODS: Positive blood cultures from 1998 to 2004 were reviewed. Patients whose cultures grew >1 organism were age-matched with monomicrobial bloodstream infection controls. Records were reviewed to compare their underlying medical conditions, organisms isolated, adequacy of therapy, and clinical characteristics of illness. RESULTS: Twenty-nine episodes of PBSI were identified in 18 subjects. PBSI patients were more likely to have chronic medical conditions, chronic gastrointestinal pathology, central venous catheters, and to be receiving parenteral nutrition than controls. Pathogens found more commonly in PBSI episodes included Enterococcus spp., coagulase negative staphylococci, and Candida spp. Empiric antimicrobial therapy was less likely to be adequate in patients with PBSI. PBSI patients were hospitalized longer, required longer intensive care and had prolonged bloodstream infection. Subjects with PBSI had prolonged duration of fever and had higher degrees of sepsis than controls. CONCLUSIONS: Chronic medical conditions, particularly gastrointestinal disease, are risk factors for PBSIs. Because clinical illness may be more severe, alteration of the empiric antimicrobial regimen should be considered in some of these patients. PMID- 18398387 TI - Acute childhood encephalitis and encephalopathy associated with influenza: a prospective 11-year review. AB - BACKGROUND: Influenza virus infection has been associated with a variety of neurologic complications. The objective of this study was to evaluate prospectively the role of influenza viruses in acute childhood encephalitis/encephalopathy (ACE). METHODS: All children admitted to the Hospital for Sick Children, Toronto, during an 11-year period with ACE and evidence of acute influenza virus infection were included. Acute influenza virus infection was defined by detection of the organism in the nasopharynx by direct immunofluorescence microscopy or viral culture and/or by a 4-fold or greater rise in complement fixation titer. RESULTS: A total of 311 children with ACE were evaluated; evidence of influenza infection was detected in 7% (22 of 311). Eight were excluded from the main analysis because of evidence implicating other potential pathogens. Eleven of the 14 included subjects were <5 years of age. A respiratory prodrome was documented in 93% of subjects. In 64% neurologic manifestations developed within 5 days of onset of respiratory symptoms. Neuroimaging abnormalities were more common in children <2 years of age. Neurologic sequelae occurred in more than one-half of subjects. CONCLUSIONS: In this prospective registry, influenza virus infection was associated with 5% of ACE cases. The majority of children were <5 years of age and the prevalence of neuroimaging abnormalities was higher in children <2 years of age suggesting that younger children are predisposed to the neurologic complications of influenza. An acute rather than a postinfectious process was suggested by the briefness of the respiratory prodrome in most cases. PMID- 18398388 TI - Prognostic factors in influenza-associated encephalopathy. AB - BACKGROUND: Recently, reports of influenza-associated encephalopathy have increased worldwide. Given the high mortality and morbidity rates attributable to this severe neurologic complication of influenza, we conducted a nationwide study in Japan to identify the prognostic factors. METHODS: We retrospectively evaluated 442 cases of influenza-associated encephalopathy that were reported to the Collaborative Study Group on Influenza-Associated Encephalopathy, which was organized by the Japanese Ministry of Health, Labor, and Welfare in collaboration with hospitals, clinics, and local pediatric practices in Japan between 1998 and 2002. The outcome for each patient was classified as either survival or death. Predictors of death were identified using logistic regression analysis. RESULTS: Four major prognostic factors for death were found to be significant by multivariate analysis (P < 0.05) in the 184 patients for whom we had complete data: elevation of aspartate aminotransferase, hyperglycemia, the presence of hematuria or proteinuria, and use of diclofenac sodium. CONCLUSIONS: We identified patients who had factors associated with a poor prognosis, and these findings might be clinically useful for the management of this illness. PMID- 18398389 TI - REACH: from registry to harmonization. PMID- 18398390 TI - Silent coronary plaque rupture. PMID- 18398391 TI - REACH: international prospective observational registry in patients at risk of atherothrombotic events. Results for the French arm at baseline and one year. AB - INTRODUCTION: Atherothrombosis is a systemic, diffuse disease associated with a high risk of cardiovascular morbidity and mortality. It is the main cause of death in Western populations, a major public health concern and its prevalence will further increase in the future. OBJECTIVES: To evaluate the rate of major vascular events at 1 year in French patients with confirmed atherothrombotic disease, recruited in the REACH international registry. METHODS: The REACH Registry has recruited 55.000 patients in 44 countries, aged at least 45 years and suffering from established atherothrombotic disease (EAD). In France, 713 investigators selected 3.514 patients with EAD between December 2003 and June 2004. Each investigator had to include 5 to 10 patients presenting after a first documented event of cerebrovascular disease (CVD), coronary artery disease (CAD) or lower limb peripheral arterial occlusive disease (PAD). The patients were followed up for 1 year with collection of major vascular events. RESULTS: Among the 3514 French patients with EAD in the REACH registry, 2.373 (68%) had documented coronary disease, 778 (22%) had an ischemic stroke and 923 (26%) had documented PAD. One quarter of CAD patients, one third of CVD patients and one half of PAD patients had another atherothrombotic disease localization. Follow-up at 1 year was documented for 3.373 patients with EAD. The 1-year event rate in patients who had EAD was a function of the number of atherothrombotic localizations: the vascular death rate was 1.8% if there was a single localization and 4.1% if there were 2 or 3 localizations, and the composite death, infarct and stroke rates were 3.8% and 7.2% respectively and 11.7% and 22.3% respectively if hospitalizations were added to the latter endpoint. CONCLUSION: The number of major vascular events during the first year is high in EAD patients although these patients were followed up on an outpatient basis and are considered to be stable. In patients with prior EAD, there was a close link between the incidence of major vascular events and the number of symptomatic arterial beds (2 or 3 sites). The risk of a major vascular event was twice as high in patients with polyvascular involvement than in those who only had one affected artery. PMID- 18398392 TI - Videoconference pediatric and congenital cardiology consultations: a new application in telemedicine. AB - BACKGROUND: The relative rarity of congenital heart disease gives it an orphean disease status, requiring specialised centres. The present maturity of information technology allows telemedicine to be integrated into current medical practice. We report our experience of telemedicine between the cardiology department at St Pierre Hospital on the island of Reunion and the pediatric cardiology department at the teaching hospital in Toulouse. AIMS: The aims of this work were to 1. verify the technical feasibility of transmitting echocardiographic images, 2. determine an optimal therapeutic strategy for each patient, and 3. deliver precise information live to patients and their families. METHODS: Five pediatric cardiology videoconference consultation sessions were transmitted between April 2006 and May 2007. The videoconference equipment, POLYCOM VSX 7000 (R), was used to relay information between the two centres, using six high-debit digital telephone lines, allowing a transfer rate of 384 kbits/s and an image frequency of 25 frames per second. The echocardiographic equipment at St Pierre Hospital was connected to the videoconference equipment by an S-VHS video output. The transmitted sources alternated between the echographic video output and the signal from a video camera, with continuous audio transmission. RESULTS: The telemedicine meeting was made up of three main elements: 1. a consultation with real-time echocardiographic acquisition and transmission, 2. a discussion between medical colleagues, and 3. a discussion with the family. Five videoconference consultation sessions were organised between April 2006 and May 2007. 22 patients were involved (median age 3 years, age range 7 days to 48 years). Heart disease was congenital in 20 patients, and acquired in 2 patients. The aim of the telemedicine consultation was to specify: 1. medical treatment in 7 patients, and 2. an indication for surgery or interventional catheterisation in 15 patients. There was no significant change in diagnosis, but in 2 patients with complex heart disease some anatomical clarifications were made. For 3 patients, the videoconference discussion was essential to get the extremely reticent families to accept the indication for surgery. CONCLUSION: This is the first experience in France of telemedicine consultation for pediatric and congenital cardiology. These videoconferences allowed patients in the south of Reunion to benefit from a specialist opinion on optimal therapeutic strategy, with no delay or need to travel a long distance. PMID- 18398393 TI - Endovascular treatment of descending aortic dissection (type B): short- and medium-term results. AB - BACKGROUND: Optimal treatment of type B dissections is open to debate. The use of endoprostheses is an option that requires evaluation. AIM: To report our experience with endoprostheses in type B aortic dissections. METHODS: We report our short- and medium-term results with covered prostheses for the treatment of acute (n=7) and chronic (n=28) type B aortic dissections. The criteria used to indicate treatment were the same as those usually used for surgery: acute complications or dilated aneurysm. Cover of the main intimal tear was obtained in all cases with an improvement in symptoms in patients with acute dissections. RESULTS: Early mortality was 14.3% (five patients), linked in three cases to the occurrence of a retrograde dissection of the ascending aorta. No neurological complications were observed. Four patients required an additional endovascular and/or surgical procedure. On early control scans, complete thrombosis of the false lumen at the thoracic level was observed in 40% of cases, partial thrombosis in 42.8% and an absence of thrombosis in 11.4%. After a mean follow-up of 20.8 months, one patient died of a pneumopathy. No secondary aneurysm expansion was noted at the thoracic stage whereas three patients presented with dilation of the abdominal aorta. CONCLUSION: The results of treatment of type B dissections with covered endoprostheses are encouraging. However, the morbimortality associated with treatment and the uncertainty of long-term results do not allow the use of this therapeutic option outside the criteria usually recognized to indicate surgery. PMID- 18398394 TI - Resistance to platelet antiaggregants: an important cause of very late thrombosis of drug eluting stents? Observations from five cases. AB - BACKGROUND: Very late thrombosis of drug eluting stents is a rare complication that might be triggered by resistance to platelet antiaggregants (PAAs). AIM: Following an initial case where clinical data strongly suggested resistance to PAAs, we carried out a prospective systematic analysis of platelet aggregation in four subsequent cases of late thrombosis. METHODS: Resistance to aspirin was investigated with the PFA-100 test employing a collagen-epinephrine cartridge (Platelet Function Analyzer; Dade Behring). Resistance to clopidogrel was determined by flow cytometry of intraplatelet vasodilator-stimulated phosphoprotein (VASP) phosphorylation. RESULTS: All four cases showed resistance to either aspirin or clopidogrel, and two cases showed dual resistance to both of these PAAs. CONCLUSION: Analysis of platelet function in a patient with late stent thrombosis is useful and may allow adaptation of subsequent patient management. The value of monitoring platelet function after implantation of a drug eluting stent should be evaluated in prospective studies. PMID- 18398395 TI - Changes in characteristics of hospitalized heart failure patients in ten years: a single-center study. AB - BACKGROUND: Progress in therapy and the ageing patients hospitalized with heart failure may have impacted the characteristics of this patients. AIMS: We compared epidemiological data of patients admitted with heart failure during two periods separated by a 10 year interval. METHODS: Characteristics of 353 heart failure patients recruited between 2002 and 2004 with those of 304 heart failure patients recruited between 1992 and 1994 were compared retrospectively. RESULTS: There is now a majority of male patients (56.4%) not found ten years ago. The average age is unchanged (75.1+/-11 then 76.4+/-11 years) even though the proportion of patients aged over 70 years has increased (75% versus 70%). Hospital length of stay has fallen from 14+/-9 to 10+/-7 days. Hospital mortality (8%) are identical. The two main etiologies remain coronary and hypertensive heart disease at 29 and 24% respectively but these proportions are lower than ten years ago (42 and 28% respectively). The ejection fraction is more often preserved (56%) than before (44%). Increased prescription of inhibitors of the renin-angiotensin system and beta-blockers is confirmed. Post-hospital and total mortality has fallen by 50 and 30% from 30 to 16% and 35 to 24% respectively at the expense of a 25% increase in the frequency of hospital readmissions from 29 to 38%. CONCLUSION: Even if mortality has declined, heart failure remains a major public health burden with a significant number of hospital readmissions. Other approaches such as therapeutic education must therefore be developed. PMID- 18398396 TI - Reliability and limitations of angiography in the diagnosis of coronary plaque rupture: an intravascular ultrasound study. AB - OBJECTIVE: Intravascular ultrasound (IVUS) is a cornerstone tool for the diagnosis of plaque rupture (PR) but is usually used secondary to the suspicion of PR on angiography; the true incidence of PR may therefore be overestimated. We sought to evaluate predictors of angiographic diagnosis of PR using a non angiographically driven IVUS examination. METHODS AND RESULTS: Diagnosis of PR on IVUS required agreement between two operators blinded to the results of angiography. Any irregular lesion with ulceration, flap or aneurysm on a qualitative angiogram was considered suspicious for PR. IVUS-detected PR and non PR lesions were compared with the corresponding angiograms. A total of 224 distinct (ruptured or non-ruptured) lesions were detected by IVUS in 65 patients; 49 of the 105 IVUS-detected non-culprit PRs were suspected on angiography. The positive and negative predictive values for correct angiographic diagnosis of PR were 96% and 61%, respectively. Proximal coronary location, wide cavity, and counterflow rupture were strong predictors of correct angiographic diagnosis, enabling four specific angiographic patterns to be identified using three dimensional IVUS PR reconstruction. CONCLUSION: Against IVUS as the gold standard, angiographic diagnosis of PR showed good specificity but low sensitivity. However, better angiographic diagnosis should enable medical treatment to be optimized, especially with respect to statin therapy. PMID- 18398397 TI - Arrhythmic sudden death in children. AB - Sudden death (SD) in childhood is rare, representing only 10% of paediatric mortality after one year of age. The individual risk is estimated between 1 in 20.000 and 1 in 50.000 per year. In case of a negative autopsy for cardiac morphologic anomalies, the most presumable cause remains a genetically-determined malignant primary ventricular arrhythmia. Rhythmic sudden cardiac death can be categorized as a complication of a cardiomyopathy (dilated or hypertrophic), or as a primary channelopathy without any structural heart disease. Primary ventricular arrhythmias include long QT syndrome, Brugada syndrome, short QT syndrome and Polymorphic Ventricular Tachycardia. The diagnosis of such syndromes relies upon specific ECG anomalies, personal history of family members, eventually echocardiography and drug challenge. For some of these diseases, morbid genes have been identified thus rendering possible the management of pre symptomatic or undiagnosed family members within specialized multidisciplinary teams. In case of sudden arrhythmic death in children, the parents and siblings must be examined Rescued sudden death exposes to a high risk of recurrence. In such patients, the automatic implantable defibrillator has dramatically improved survival. PMID- 18398398 TI - Transcatheter aortic valve implantation: technical aspects, results and indications. AB - The development of the percutaneous heart valve (PHV) may become a primary therapeutic modality for the high risk and inoperable patients with critical symptomatic aortic stenosis. The first human percutaneous aortic valve implant was performed by our group in April 2002. To date, more than 500 Cribier-Edwards PHV have been implanted worldwide using arterial trans-femoral or trans-apical approach. Data on the retrograde transfemoral approach is growing with more than 270 patients implanted as of October 2007. Procedural success rate is high (86%) and the 30-day mortality is 12%. Today, 2 patients are alive at a follow-up of more than 4 years. The same Cribier-Edwards-PHV can be implanted using trans apical approach. In this procedure, PHV is introduced under direct vision into the left ventricle via a mini-thoracotomy. This obviates the concerns regarding vascular access in the presence of small caliber vessels and/or vascular occlusive disease. More than 200 patients have been treated with this approach. In the European experience 30-day mortality is 14%. There is intense interest in PHV technology, and there are multiple devices at various stages of development in animals and humans. The most developed is the CoreValve Revalving Technology. More than 350 patients have been treated with this technique. The immediate and mid-term results with this device are promising with a procedural success of 92% and a 30-day mortality of 15%. The future of this technology and its application is dependent on the continued collaboration between general internists, cardiologists, surgeons, engineers, and industry. PMID- 18398399 TI - Demonstration of a complex coronary fistula between the left anterior descending coronary artery (LAD) and the pulmonary artery using multislice CT. PMID- 18398400 TI - Ventricular aneurysm versus pseudoaneurysm: role of multi-imaging modality. PMID- 18398401 TI - Should patients with implantable cardioverter-defibrillators refrain from driving? PMID- 18398402 TI - Evaluation of a screening protocol using auscultation and portable echocardiography to detect asymptomatic rheumatic heart disease in Tongan schoolchildren. AB - BACKGROUND: Rheumatic heart disease (RHD) is an important problem in developing countries; however, many cases are detected only when the disease has progressed to cardiac failure. Screening can detect cases earlier, but there are no screening guidelines. METHODS: We performed a cross-sectional screening study in Tonga among 5,053 primary school children, in whom auscultation followed by echocardiography of those with heart murmurs were used to identify RHD. We also analyzed whether a three-stage screening protocol of auscultation performed by a medical student to detect any heart murmur, second-stage auscultation performed by a local pediatrician to differentiate pathological from innocent murmurs and echocardiography of those with pathological murmurs altered outcomes. RESULTS: The prevalence of definite RHD was 33.2 per 1,000. The prevalence of RHD increased significantly with age, peaking at 42.6 per 1,000 in children aged 10 12 years. Most valve lesions (91 [54%] of 169) were mild. Auscultation to detect pathological murmurs was poorly sensitive (46.4%), and the finding of any murmur on auscultation did not affect the likelihood of detecting pathology on echocardiography. The finding of a pathological murmur did significantly increase the likelihood of detecting pathology on echocardiography, but still missed 54% of those with pathology (mainly RHD) detected on echocardiography. CONCLUSIONS: Screening is a useful method for detecting asymptomatic RHD in regions of high prevalence and we report a high echocardiographically confirmed prevalence. The most appropriate screening strategy remains to be confirmed, however, and implementation will depend on the availability of echocardiography and trained staff. PMID- 18398403 TI - Once-yearly zoledronate-an effective preventative therapy for new fractures after hip fracture? PMID- 18398404 TI - Use of plasma chromogranin A and urine fractionated metanephrines to diagnose pheochromocytoma? PMID- 18398405 TI - Does high-trauma fracture increase the risk of subsequent osteoporotic fracture? PMID- 18398406 TI - Management of advanced germ-cell tumors of the testis. AB - Advanced tumors of the testis are curable. Standard treatment includes chemotherapy with a combination of bleomycin, etoposide and cisplatin, followed by surgical resection of residual tumor. The number of cycles of chemotherapy needed depends on prognostic factors such as the primary site, histology, presence of visceral metastases, and serum levels of tumor markers. Patients with a favorable risk profile receive three cycles of chemotherapy, and those with increased risk receive four cycles. After chemotherapy, resection of all residual local disease and systematic retroperitoneal dissection of bulky lymph-node disease are mandatory for patients with nonseminoma germ-cell tumors. In patients with seminoma, surgery is required when residual disease is either bulky or functional on (18)fluorodeoxyglucose-PET scan. When complete resection of necrosis, teratoma and/or active germ-cell cancer has been done, no further treatment is needed. The consequences of therapy are complex: treatment could affect fertility, sexuality, metabolic status and renal and neurological function. Secondary malignancies are reported, as well as contralateral germ-cell tumors. Owing to the complexity of treatment and the multidisciplinary approach required, patients with advanced germ-cell tumors should be managed in high volume centers with experience of treating large numbers of patients. PMID- 18398407 TI - Contralateral intratubular germ cell neoplasia in a patient with testicular cancer. AB - BACKGROUND: A 27-year-old man was referred to an oncology department following right orchiectomy for a stage I testicular seminoma at high risk for recurrence. He presented 6 weeks after the orchiectomy with an atrophic left testis, fatigue and a history of infertility. INVESTIGATIONS: Measurement of serum levels of urea, electrolytes, liver enzymes, bilirubin, human chorionic gonadotropin, alpha fetoprotein, lactate dehydrogenase, testosterone and luteinizing hormone, full blood count, and left testicular biopsy. DIAGNOSIS: Tubular atrophy of the left testis with islands of intratubular germ cell neoplasia (ITGCN), and hypergonadotropic hypogonadism. MANAGEMENT: The patient received adjuvant chemotherapy as a single dose of carboplatin for the seminoma at high risk for recurrence, and testosterone replacement for the hypergonadotropic hypogonadism. Radiotherapy to the ITGCN-bearing solitary testis or a second orchiectomy was offered to prevent the progression of ITGCN into an invasive germ cell tumor. After exploring his options with regards to fertility treatment, the patient chose to undergo second orchiectomy with a subsequent, unsuccessful, attempt at sperm retrieval. At 20 months after diagnosis of his initial seminoma the patient showed no sign of recurrence. PMID- 18398408 TI - What is the optimal anti-inflammatory therapy for patients with osteoarthritis and increased cardiovascular risk? PMID- 18398409 TI - Infliximab-induced disseminated histoplasmosis in a patient with Crohn's disease. AB - BACKGROUND: A 56-year-old female with a 30-year history of ileocolic Crohn's disease presented with a 1-month history of bloody diarrhea and decreased caliber of stools; physical examination revealed a broad indurated anal fissure. The patient had been receiving antimetabolite therapy with 6-mercaptopurine and maintenance therapy with infliximab for over a year. INVESTIGATIONS: Physical examination; proctoscopy; perianal and anal canal biopsy; chest CT; blood and stool analysis, measurement of serum histoplasmosis antibodies and urine histoplasmosis antigen levels; fungal culture and Gomori's methenamine silver staining of resected tissue specimens. DIAGNOSIS: Disseminated histoplasmosis. MANAGEMENT: Proctocolectomy and end ileostomy followed by treatment with liposomal amphotericin and then oral itraconazole. A palmar space abscess required multiple debridements, and a muscle flap to cover the defect. PMID- 18398410 TI - Does acupuncture improve symptoms in patients with osteoarthritis who are awaiting knee replacement surgery? PMID- 18398411 TI - Drug Insight: choosing a drug treatment strategy for women with osteoporosis-an evidence--based clinical perspective. AB - Many randomized controlled trials (RCTs) have investigated drug treatment for women at high risk of fracture, with a reduction in fracture risk as their end point. There has also been progress in identifying women at the highest risk of fractures. The most important clinical determinant contributing to the clinical decision of initiating and choosing drug therapy for fracture prevention is a woman's fracture risk, which, in RCTs, was determined by menopausal state, age, bone mineral density, fracture history, fall risks and glucocorticoid use. Women with secondary osteoporosis were excluded, except in studies of glucocorticoid use. A second determinant of drug therapy is the evidence for fracture prevention in terms of spectrum (vertebral, nonvertebral and/or hip fractures), size and speed of effect. In the absence of head-to-head RCTs with fracture risk as the end point, however, the efficacy of antifracture drugs cannot be directly compared. Other determinants include the potential extraskeletal benefits and safety concerns of the drug, patient preferences and reimbursement issues. PMID- 18398412 TI - Cardiorenal and renocardiac syndromes: the need for a comprehensive classification and consensus. PMID- 18398413 TI - Early steroid treatment for drug-induced acute interstitial nephritis. PMID- 18398414 TI - Transplantation of allogeneic hematopoietic stem cells: an emerging treatment modality for solid tumors. AB - Allogeneic transplantation of hematopoietic cells from an HLA-compatible donor has been used to treat hematologic malignancies. Allogeneic transplantation not only replaces the marrow affected by the disease, but exerts an immune graft versus-tumor (GVT) effect mediated by donor lymphocytes. The development of nonmyeloablative conditioning regimens before allogeneic transplantation has allowed this therapy to be used in elderly and disabled patients. An allogeneic GVT effect is observed in a proportion of patients with renal, breast, colorectal, ovarian, and pancreatic cancer treated with allogeneic transplantation. In general, the tumor response is associated with the development of acute and chronic graft-versus-host disease. Further improvements will depend on the identification of the antigen targets of GVT, and on reduction of the toxicity of the procedure. Targeted therapies may complement the immune effect of allogeneic transplantation. We present updated results from the literature and data recently placed on file at the European Bone Marrow Transplantation Solid Tumors Working Party. PMID- 18398415 TI - Sleep disturbances in patients with parkinsonism. AB - Altered sleep and vigilance are among the most frequent symptoms, besides parkinsonism, in movement disorders. As many as 60% of patients with Parkinson's disease (PD) experience insomnia, 15-59% show rapid eye movement (REM) sleep behavior disorders (RBDs), and 30% show excessive daytime sleepiness. Insomnia is a distressing difficulty to maintain sleep, which is exacerbated by motor disability, painful dystonia, restless legs, dysuria, anxiety and depressed mood. Improving night-time motor control by overnight treatment with levodopa, transdermal or long-acting dopamine agonists, or bilateral subthalamus stimulation, can improve sleep continuity. RBDs are violent, enacted dreams that expose the patient or their sleeping partner to night-time injuries. A striking improvement of parkinsonism is observed during these behaviors in PD. RBDs are thought to be caused by lesions in the REM sleep atonia system, and can, in association with other early markers of neurodegenerative diseases, such as olfactory, cognitive and autonomic disturbances, precede parkinsonism by several years. Daytime sleepiness, often with a narcolepsy-like phenotype, is a common occurrence in PD, owing to lesions in the arousal systems of the brain. The use of dopamine agonists increases the risk of sleep attacks, especially when driving, suggesting a drug-disease interaction. PMID- 18398416 TI - Preconditioning mediated by sublethal oxygen-glucose deprivation-induced cyclooxygenase-2 expression via the signal transducers and activators of transcription 3 phosphorylation. AB - The signal transducers and activators of transcription (STATs) were found to be essential for cardioprotection. However, their role in preconditioning (PC) neuroprotection remains undefined. Previously, our studies showed that PC mediated a signaling cascade that involves activation of epsilon protein kinase C (varepsilonPKC), extracellular signal-regulated kinase (ERK1/2), and cyclooxygenase-2 (COX-2) pathways. However, the intermediate pathway by which ERK1/2 activates COX-2 was not defined. In this study, we investigated whether the PC-induced signaling pathway requires phosphorylation of STAT isoforms for COX-2 expression. To mimic PC or lethal ischemia, mixed cortical neuron/astrocyte cell cultures were subjected to 1 and/or 4 h of oxygen-glucose deprivation (OGD), respectively. The results indicated serine phosphorylation of STAT3 after PC or varepsilonPKC activation. Inhibition of either varepsilonPKC or ERK1/2 activation abolished PC-induced serine phosphorylation of STAT3. Additionally, inhibition of STAT3 prevented PC-induced COX-2 expression and neuroprotection against OGD. Therefore, our findings suggest that PC signaling cascade involves STAT3 activation after varepsilonPKC and ERK1/2 activation. Finally, we show that STAT3 activation mediates COX-2 expression and ischemic tolerance. PMID- 18398417 TI - Arctic ground squirrel (Spermophilus parryii) hippocampal neurons tolerate prolonged oxygen-glucose deprivation and maintain baseline ERK1/2 and JNK activation despite drastic ATP loss. AB - Oxygen-glucose deprivation (OGD) initiates a cascade of intracellular responses that culminates in cell death in sensitive species. Neurons from Arctic ground squirrels (AGS), a hibernating species, tolerate OGD in vitro and global ischemia in vivo independent of temperature or torpor. Regulation of energy stores and activation of mitogen-activated protein kinase (MAPK) signaling pathways can regulate neuronal survival. We used acute hippocampal slices to investigate the role of ATP stores and extracellular signal-regulated kinase (ERK)1/2 and Jun NH(2)-terminal kinase (JNK) MAPKs in promoting survival. Acute hippocampal slices from AGS tolerated 30 mins of OGD and showed a small but significant increase in cell death with 2 h OGD at 37 degrees C. This tolerance is independent of hibernation state or season. Neurons from AGS survive OGD despite rapid ATP depletion by 3 mins in interbout euthermic AGS and 10 mins in hibernating AGS. Oxygen-glucose deprivation does not induce JNK activation in AGS and baseline ERK1/2 and JNK activation is maintained even after drastic depletion of ATP. Surprisingly, inhibition of ERK1/2 or JNK during OGD had no effect on survival, whereas inhibition of JNK increased cell death during normoxia. Thus, protective mechanisms promoting tolerance to OGD by AGS are downstream from ATP loss and are independent of hibernation state or season. Journal of Cerebral Blood Flow & Metabolism (2008) 28, 1307-1319; doi:10.1038/jcbfm.2008.20; published online 9 April 2008. PMID- 18398418 TI - Genome-wide association studies for complex traits: consensus, uncertainty and challenges. AB - The past year has witnessed substantial advances in understanding the genetic basis of many common phenotypes of biomedical importance. These advances have been the result of systematic, well-powered, genome-wide surveys exploring the relationships between common sequence variation and disease predisposition. This approach has revealed over 50 disease-susceptibility loci and has provided insights into the allelic architecture of multifactorial traits. At the same time, much has been learned about the successful prosecution of association studies on such a scale. This Review highlights the knowledge gained, defines areas of emerging consensus, and describes the challenges that remain as researchers seek to obtain more complete descriptions of the susceptibility architecture of biomedical traits of interest and to translate the information gathered into improvements in clinical management. PMID- 18398419 TI - Genetic programming of liver and pancreas progenitors: lessons for stem-cell differentiation. AB - The liver and pancreas arise from a common multipotent population of endoderm cells and share many aspects of their early development. Yet each tissue originates from multiple spatial domains of the endoderm, under the influence of different genes and inductive cues, and obtains different regenerative capacities. Emerging genetic evidence is illuminating the ability of newly specified hepatic and pancreatic progenitors to reverse their course and develop into gut progenitors. Understanding how tissue programming can be reversed and how intrinsic regenerative capacities are determined should facilitate the discovery of the basis of cellular plasticity and aid in the targeted programming and growth of stem cells. PMID- 18398420 TI - Effects of the 'live high-train low' method on prooxidant/antioxidant balance on elite athletes. AB - BACKGROUND/OBJECTIVES: We previously demonstrated that acute exposure to hypoxia (3 h at 3000 m) increased oxidative stress markers. Thus, by using the 'living high-training low' (LHTL) method, we further hypothesized that intermittent hypoxia associated with endurance training alters the prooxidant/antioxidant balance. SUBJECTS/METHODS: Twelve elite athletes from the Athletic French Federation were subjected to 18-day endurance training. They were divided into two groups: one group (control group) trained at 1200 m and lived in hypoxia (2500-3000 m simulated altitude) and the second group trained and lived at 1200 m. The subjects performed an acute hypoxic test (10 min at 4800 m) before and immediately after the training. Plasma levels of advanced oxidation protein products (AOPP), malondialdehydes (MDA), ferric-reducing antioxidant power (FRAP), lipid-soluble antioxidants normalized for triacylglycerols, and cholesterol and retinol were measured before and after the 4800 m tests. RESULTS: After the training, MDA and AOPP concentrations were decreased in response to the 4800 m test only for the control group. Eighteen days of LHTL induced a significant decrease of all antioxidant markers (FRAP, P=0.01; alpha-tocopherol, P=0.04; beta-carotene, P=0.01 and lycopene, P=0.02) for the runners. This imbalance between antioxidant and prooxidant might result from insufficient intakes in vitamins A and E. CONCLUSIONS: The LHTL model characterized by the association of aerobic exercises and intermittent resting hypoxia exposures decreased the antioxidant status whereas the normoxic endurance training induced preconditioning mechanisms in response to the 4800 m test. PMID- 18398421 TI - Presence of 25(OH)D deficiency and its effect on vitamin D receptor mRNA expression. AB - BACKGROUND AND OBJECTIVES: Vitamin D and its metabolites act through vitamin D receptor (VDR). We hypothesized that subjects with low serum 25(OH)D levels but normal PTH might have increased VDR expression. DESIGN AND METHODS: VDRmRNA expression was assessed by real time PCR in duodenal mucosa and PBMC (peripheral blood mononuclear cells) in 45 subjects with normal duodenoscopy and in PBMC alone in 48 healthy volunteers with hypovitaminosis D. 25(OH)D, PTH and VDRmRNA expression in PBMC was reassessed after 8 weeks of oral cholecalciferol (60 000 IU per week) in a subset (n=23) of healthy volunteers. RESULTS AND CONCLUSIONS: The VDRmRNA expressions in the duodenum and PBMC were significantly correlated (r=0.42), but the expression was 13 times higher in the former than the latter. The mean VDRmRNA expression was similar in 25(OH)D-deficient subjects with or without PTH elevation, both in the duodenum and PBMC. The PBMC VDRmRNA expression showed no significant change after cholecalciferol supplementation. A weak correlation coefficient between duodenal mucosa and PBMC VDRmRNA suggests that caution needs to be exercised while using the latter as a surrogate for other sites. PMID- 18398422 TI - Factor VII, blood lipids and fat intake: gene-nutrient interaction and risk of coronary heart disease with the factor VII R353Q polymorphism. AB - BACKGROUND: The relation between dietary fat, blood lipids, plasma factor VII coagulant activity (FVIIc) and risk of coronary heart disease (CHD) according to the R353Q polymorphism in the factor VII gene was assessed. METHODS: Cross sectional study of 15,073 individuals participating in the European Prospective Investigation of Cancer (EPIC) Norfolk, 7433 of which had FVIIc available. Nested case-control study of 985 CHD cases and 2009 matched controls. RESULTS: FVIIc was significantly associated with total fat intake in females, especially in the RR homozygotes (standardized beta=0.24; 95% confidence interval (95% CI) 0.08-0.40; P<0.01), but there were no associations with intakes of saturated, monounsaturated or polyunsaturated fatty acids according to genotype and no associations in males. FVIIc was significantly positively associated with total cholesterol (P<0.01) and with triacylglycerol (P<0.001) in both genders, with an interaction according to genotype for triacylglycerol in males: beta Q allele carriers 0.26 (95% CI 0.18-0.34), beta RR homozygotes 0.16 (95% CI 0.12-0.20) (Z interaction=-2.24; P<0.05). There was no effect of genotype on the odds ratio (OR) for incident CHD: OR 0.89 for Q allele carriers compared with RR homozygotes (95% CI 0.77-1.02) in 985 cases and 2009 matched controls. CONCLUSION: These results show a strong association between dietary fat intake and FVIIc in women, and between serum triacylglycerol and cholesterol and FVIIc levels in both genders. The R353Q genotype only marginally affected modulation of FVIIc by dietary fat. The association between triacylglycerol and FVIIc was significantly stronger in males carrying the Q allele than in those with the RR genotype. PMID- 18398423 TI - Moderate alcohol consumption the night before glycaemic index testing has no effect on glycaemic response. AB - A randomized crossover study was carried out to determine whether moderate alcohol consumption the night preceding glycaemic index testing has an effect on glycaemic response. Test foods consisted of two glucose beverages and two bread samples, containing 50 g of available carbohydrate. Each test food was consumed with and without alcohol the night preceding testing. Glucose concentration was measured over 2 h and area under the curve (AUC) determined. The AUC for bread and glucose without and with alcohol did not differ for either the glucose drink (ratio (95% confidence intervals): 1.00 (0.8, 1.1)) or bread (0.9; (0.8, 1.1)). Further, there was no difference in the glycaemic index (GI) of the bread with and without alcohol (64 (6) vs 67 (2)%; P=0.78). Moderate alcohol consumption the evening before GI testing does not affect glycaemic response or GI determination. PMID- 18398424 TI - Heterozygosity and lungworm burden in harbour seals (Phoca vitulina). AB - In several studies, heterozygosity measured at around 10 microsatellite markers correlates with parasite load. Usually the effect size is small, but while this may reflect reality, it may also be possible that too few markers are used or the measure of fitness contains too much error to reveal what is actually a much stronger underlying effect. Here, we analysed over 200 stranded harbour seals (Phoca vitulina) for an association between lungworm burden and heterozygosity, conducting thorough necropsies on the seals and genotyping the samples obtained for 27 microsatellites. We found that homozygosity predicts higher worm burdens, but only in young animals, where the worms have the greatest impact on fitness. Testing each locus separately, we found that a significant majority reveal a weak but similar trend for heterozygosity to be protective against high lungworm burden, suggesting a genome-wide effect, that is, inbreeding. This conclusion is supported by the fact that heterozygosity is correlated among markers in young animals but not in otherwise equivalent older ones. Taken as a whole, our results support the notion that homozygosity increases susceptibility to parasitic infection and suggest that parasites can be effective in removing inbred individuals from the population. PMID- 18398425 TI - DNA barcoding plants in biodiversity hot spots: progress and outstanding questions. PMID- 18398427 TI - Rapid lentiviral transduction preserves the engraftment potential of Fanca(-/-) hematopoietic stem cells. AB - Fanconi anemia (FA) is a rare recessive syndrome, characterized by congenital anomalies, bone marrow failure, and predisposition to cancer. Two earlier clinical trials utilizing gamma-retroviral vectors for the transduction of autologous FA hematopoietic stem cells (HSCs) required extensive in vitro manipulation and failed to achieve detectable long-term engraftment of transduced HSCs. As a strategy for minimizing ex vivo manipulation, we investigated the use of a "rapid" lentiviral transduction protocol in a murine Fanca(-/-) model. Importantly, while this and most murine models of FA fail to completely mimic the human hematopoietic phenotype, we observed a high incidence of HSC transplant engraftment failure and low donor chimerism after conventional transduction (CT) of Fanca(-/-) donor cells. In contrast, rapid transduction (RT) of Fanca(-/-) HSCs preserved engraftment to the level achieved in wild-type cells, resulting in long-term multilineage engraftment of gene-modified cells. We also demonstrate the correction of the characteristic hypersensitivity of FA cells against the cross-linking agent mitomycin C (MMC), and provide evidence for the advantage of using pharmacoselection as a means of further increasing gene-modified cells after RT. Collectively, these data support the use of rapid lentiviral transduction for gene therapy in FA. PMID- 18398426 TI - HSP70 and constitutively active HSF1 mediate protection against CDCrel-1-mediated toxicity. AB - Defects in cellular quality control mechanisms are thought to contribute to the neuropathology of Parkinson's disease (PD). Overexpressing heat shock proteins (HSPs) may constitute a powerful therapeutic strategy for PD, because they boost the ability of the cell to eliminate unwanted proteins. We investigated the neuroprotective potential of HSP70, HSP40, and H-BH, a constitutively active form of heat shock factor 1, in a rat model of PD based on adeno-associated virus (AAV) vector-mediated overexpression of CDCrel-1, a parkin substrate known to be toxic to dopaminergic neurons. AAV vector-mediated overexpression of H-BH and of HSP70 afforded similar levels of protection against CDCrel-1 toxicity, with approximately 20% improvement in survival of dopaminergic neurons as compared to the controls. The assessment of protection conferred was made using tyrosine hydroxylase (TH) and HuC/D immunohistochemistry and Fluoro-Gold retrograde tracing, and by observing the extent of preservation of spontaneous function and also the extent of drug-induced motor function. In contrast to H-BH and HSP70, HSP40 overexpression exacerbated CDCrel-1-mediated cell death. Real-time reverse transcriptase (RT)-PCR analysis showed that H-BH had the effect of upregulating endogenous HSP70 and HSP40 mRNA levels 10-fold and 4-fold over basal levels, respectively, whereas AAV vector-mediated HSP70 and HSP40 mRNA levels were over 100-fold higher. Our results suggest that a comparatively modest upregulation of multiple HSPs may be an effective approach for achieving significant neuroprotection in PD. PMID- 18398428 TI - Computer-assisted hydrodynamic gene delivery. AB - The recently developed hydrodynamic delivery method makes it possible to deliver DNA and RNA into parenchyma cells by intravascular injection of nucleic acid containing solution. While this procedure is effective in rodents, it is difficult to perform in large animals, because manual control while delivering the injection cannot be sufficiently reliable for achieving a just-right hydrodynamic pressure in targeted tissue. In order to overcome this problem, we have developed a computer-controlled injection device that uses real-time intravascular pressure as a regulator. Using the new injection device, and mouse liver as the model organ, we demonstrated continuous injection at a single pressure and different pressures, and also serial (repeated) injections at intervals of 250 ms, by programming the computer according to the need. When assessed by reporter plasmids, the computer-controlled injection device exhibits gene delivery efficiency similar to that of conventional hydrodynamic injection. The device is also effective in gene delivery to kidney and muscle cells in rats, with plasmids or adenoviral vectors as gene carriers. Successful gene delivery to liver and kidney was also demonstrated in pigs, with the computer-controlled injection being combined with image-guided catheterization. These results represent a significant advance in in vivo gene delivery research, with potential for use in gene therapy in humans. PMID- 18398429 TI - Novel cancer antiangiotherapy using the VEGF promoter-targeted artificial zinc finger protein and oncolytic adenovirus. AB - Inhibition of tumor angiogenesis through modulation of vascular endothelial growth factor (VEGF) and its signaling pathway has been clinically validated as a viable therapeutic modality in the treatment of cancer. The use of artificial transcription factors based on Cys2-His2 zinc-finger proteins (ZFPs) targeting the VEGF promoter offers a novel strategy for modulating VEGF levels in tumors. In order to demonstrate the utility of VEGF-targeted ZFPs as therapeutic agents, we generated adenoviruses (Ads) expressing VEGF promoter-targeted transcriptional repressor ZFP, F435-KOX. A replication-incompetent Ad expressing F435-KO X, namely, Ad-DeltaE1-KOX, significantly reduced VEGF expression and functionally led to inhibition of angiogenesis. In vivo, an oncolytic Ad expressing F435-KOX, namely, Ad-DeltaB7-KOX, elicited a pronounced antitumor effect against a human glioblastoma xenograft model, U87MG. Further, consistent with its expected mechanism of action, Ad-DeltaB7-KOX was shown to greatly reduce the level of VEGF and vessel density in tumor tissue and increase terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)-positive apoptotic cells in tumors. Survival rates were also significantly increased in Ad-DeltaB7-KOX treated mice. Taken together, the findings from this study identify F435-KOX as a novel and potent ZFP transcription factor that can inhibit VEGF-A-mediated angiogenesis and offer a novel therapeutic modality in the treatment of cancer. PMID- 18398430 TI - Evaluation of HapMap data in six populations of European descent. AB - We studied how well the European CEU samples used in the Haplotype Mapping Project (HapMap) represent five European populations by analyzing nuclear family samples from the Swedish, Finnish, Dutch, British and Australian (European ancestry) populations. The number of samples from each population (about 30 parent-offspring trios) was similar to that in the HapMap sample sets. A panel of 186 single nucleotide polymorphisms (SNPs) distributed over the 1.5 Mb region of the GRID2 gene on chromosome 4 was genotyped. The genotype data were compared pair-wise between the HapMap sample and the other population samples. Principal component analysis (PCA) was used to cluster the data from different populations with respect to allele frequencies and to define the markers responsible for observed variance. The only sample with detectable differences in allele frequencies was that from Kuusamo, Finland. This sample also separated from the others, including the other Finnish sample, in the PCA analysis. A set of tagSNPs was defined based on the HapMap data and applied to the samples. The tagSNPs were found to capture the genetic variation in the analyzed region at r(2)>0.8 at levels ranging from 95% in the Kuusamo sample to 87% in the Australian sample. To capture the maximal genetic variation in the region, the Kuusamo, HapMap and Australian samples required 58, 63 and 73 native tagSNPs, respectively. The HapMap CEU sample represents the European samples well for tagSNP selection, with some caution regarding estimation of allele frequencies in the Finnish Kuusamo sample, and a slight reduction in tagging efficiency in the Australian sample. PMID- 18398431 TI - Maternal genotype effects can alias case genotype effects in case-control studies. PMID- 18398433 TI - Reconstructing the phylogeny of African mitochondrial DNA lineages in Slavs. AB - To elucidate the origin of African-specific mtDNA lineages, revealed previously in Slavonic populations (at frequency of about 0.4%), we completely sequenced eight African genomes belonging to haplogroups L1b, L2a, L3b, L3d and M1 gathered from Russians, Czechs, Slovaks and Poles. Results of phylogeographic analysis suggest that at least part of the African mtDNA lineages found in Slavs (such as L1b, L3b1, L3d) appears to be of West African origin, testifying to an opportunity of their occurrence as a result of migrations to Eastern Europe through Iberia. However, a prehistoric introgression of African mtDNA lineages into Eastern Europe (approximately 10 000 years ago) seems to be probable only for European-specific subclade L2a1a, defined by coding region mutations at positions 6722 and 12903 and detected in Czechs and Slovaks. Further studies of the nature of African admixture in gene pools of Europeans require the essential enlargement of databases of African complete mitochondrial genomes. PMID- 18398432 TI - Genome-wide linkage scan for atypical nevi in p16-Leiden melanoma families. AB - In most Dutch melanoma families, a founder deletion in the melanoma susceptibility gene CDKN2A (which encodes p16) is present. This founder deletion (p16-Leiden) accounts for a significant proportion of the increased melanoma risk. However, it does not account for the Atypical Nevus (AN) phenotype that segregates in both p16-Leiden carriers and non-carriers. The AN-affected p16 Leiden family members are therefore a unique valuable resource for unraveling the genetic etiology of the AN phenotype, which is considered both a risk factor and a precursor lesion for melanoma. In this study, we performed a genome-wide scan for linkage in four p16-Leiden melanoma pedigrees, classifying family members with five or more AN as affected. The strongest evidence for an atypical nevus susceptibility gene was mapped to chromosome band 7q21.3 (two-point LOD score=2.751), a region containing candidate gene CDK6. PMID- 18398434 TI - Influence of the cystathionine beta-synthase 844ins68 and methylenetetrahydrofolate reductase 677C>T polymorphisms on folate and homocysteine concentrations. AB - A high homocysteine, low folate phenotype is a feature of many diseases. The effect of the cystathionine beta-synthase (CBS) 844ins68 polymorphism on homocysteine and folate concentrations was examined alone and in the context of the 5,10-methylenetetrahydrofolate reductase (MTHFR) 677C>T polymorphism in a Northwestern European male population. The MTHFR 677TT genotype is known to be associated with increased homocysteine and decreased folate relative to CT heterozygotes and CC homozygotes in this and other populations. MTHFR 677TT homozygotes who were also CBS 844ins68 carriers had homocysteine and folate concentrations similar to those of individuals with the MTHFR 677CT and CC genotypes. Homocysteine levels in MTHFR 677TT subjects carrying the CBS 844ins68 allele were 24.1% lower than in non-carriers (6.66 vs 8.77 micromol/l, P=0.045), and serum folate levels were 27.7% higher (11.16 vs 8.74 nmol/l, P=0.034). These findings suggest that the CBS 844ins68 allele 'normalizes' homocysteine and folate levels in MTHFR 677TT individuals. PMID- 18398435 TI - Williams-Beuren syndrome TRIM50 encodes an E3 ubiquitin ligase. AB - Williams-Beuren syndrome (WBS) is a neurodevelopmental and multisystemic disease that results from hemizygosity of approximately 25 genes mapping to chromosomal region 7q11.23. We report here the preliminary description of eight novel genes mapping within the WBS critical region and/or its syntenic mouse region. Three of these genes, TRIM50, TRIM73 and TRIM74, belong to the TRIpartite motif gene family, members of which were shown to be associated to several human genetic diseases. We describe the preliminary functional characterization of these genes and show that Trim50 encodes an E3 ubiquitin ligase, opening the interesting hypothesis that the ubiquitin-mediated proteasome pathway might be involved in the WBS phenotype. PMID- 18398436 TI - MCT8 mutation analysis and identification of the first female with Allan-Herndon Dudley syndrome due to loss of MCT8 expression. AB - Mutations in the thyroid monocarboxylate transporter 8 gene (MCT8/SLC16A2) have been reported to result in X-linked mental retardation (XLMR) in patients with clinical features of the Allan-Herndon-Dudley syndrome (AHDS). We performed MCT8 mutation analysis including 13 XLMR families with LOD scores >2.0, 401 male MR sibships and 47 sporadic male patients with AHDS-like clinical features. One nonsense mutation (c.629insA) and two missense changes (c.1A>T and c.1673G>A) were identified. Consistent with previous reports on MCT8 missense changes, the patient with c.1673G>A showed elevated serum T3 level. The c.1A>T change in another patient affects a putative translation start codon, but the same change was present in his healthy brother. In addition normal serum T3 levels were present, suggesting that the c.1A>T (NM_006517) variation is not responsible for the MR phenotype but indicates that MCT8 translation likely starts with a methionine at position p.75. Moreover, we characterized a de novo translocation t(X;9)(q13.2;p24) in a female patient with full blown AHDS clinical features including elevated serum T3 levels. The MCT8 gene was disrupted at the X breakpoint. A complete loss of MCT8 expression was observed in a fibroblast cell line derived from this patient because of unfavorable nonrandom X-inactivation. Taken together, these data indicate that MCT8 mutations are not common in non AHDS MR patients yet they support that elevated serum T3 levels can be indicative for AHDS and that AHDS clinical features can be present in female MCT8 mutation carriers whenever there is unfavorable nonrandom X-inactivation. PMID- 18398437 TI - A homoplasmic mtDNA variant can influence the phenotype of the pathogenic m.7472Cins MTTS1 mutation: are two mutations better than one? AB - Mutations in mitochondrial tRNA (mt-tRNA) genes are well recognized as a common cause of human disease, exhibiting a significant degree of clinical heterogeneity. While these differences are explicable, in part, by differences in the innate pathogenicity of the mutation, its distribution and abundance, other factors, including nuclear genetic background, mitochondrial DNA (mtDNA) haplotype and additional mtDNA mutations may influence the expression of mt-tRNA mutations. We describe the clinical, biochemical and molecular findings in a family with progressive myopathy, deafness and diabetes and striking respiratory chain abnormalities due to a well-characterized heteroplasmic mt-tRNA mutation in the mt-tRNA(Ser(UCN)) (MTTS1) gene. In addition to the m.7472Cins mutation, all individuals were homoplasmic for another variant, m.7472A > C, affecting the adjacent nucleotide in the mt-tRNA(Ser(UCN)) structure. In addition to available patient tissues, we have analysed transmitochondrial cybrid clones harbouring homoplasmic levels of m.7472A > C and varying levels of the m.7472Cins mutation in an attempt to clarify the precise role of the m.7472A > C transversion in the underlying respiratory chain abnormality. Evidence from both in vivo and in vitro studies demonstrate that the m.7472A > C is able to modify the expression of the m.7472Cins mutation and would suggest that it is not a neutral variant but appears to cause a biochemical defect by itself, confirming that homoplasmic mtDNA variants can modulate the phenotypic expression of pathogenic, heteroplasmic mtDNA mutations. PMID- 18398438 TI - Preferential reciprocal transfer of paternal/maternal DLK1 alleles to obese children: first evidence of polar overdominance in humans. AB - DLK1 is part of the Notch signalling pathway that controls various developmental processes. A functional role for DLK1 in adipogenesis is suggested by several animal models. Interestingly, the DLK1 gene is imprinted in eutherian mammals. To study whether variations in DLK1 affect body weight in humans, we analysed 32 polymorphisms in a 109 kb genomic region encompassing DLK1 on human chromosome 14. In a study sample of 1025 French and German trio families comprised of both parents and extremely obese offspring we found a single nucleotide polymorphism (rs1802710) associated with child and adolescent obesity. Analysis of the allelic transmission pattern indicated the existence of polar overdominance, an unusual mode of non-Mendelian inheritance in humans previously known from the callipyge mutation in sheep. PMID- 18398439 TI - The human pseudoautosomal regions: a review for genetic epidemiologists. AB - Two intervals of sequence identity at the tips of X and Y chromosomes, the human pseudoautosomal regions PAR1 and PAR2, have drawn interest from researchers in human genetics, cytogenetics, and evolutionary biology. However, they have been widely ignored in linkage and association studies. The pseudoautosomal regions (PARs) pair and recombine during meiosis like autosomes, but the recombination activity in PAR1 is extremely different between sexes. In men, it exhibits the highest recombination frequencies of the genome. Conflicting genetic maps of this region have been estimated by using three-generation pedigrees, sperm typing, and by using haplotypes from single nucleotide polymorphisms. Male genetic map lengths in the literature vary, and linkage disequilibrium has not been analyzed in detail. We review existing tools like genetic and physical maps, linkage disequilibrium methods, linkage and association analysis, implemented statistical methods, and their suitability for PARs. For multipoint linkage analysis, sex specificity must be indicated twice, first using sex-specific maps, and second by considering the sex-specific pseudoautosomal inheritance pattern. Currently, microsatellite panels and single nucleotide polymorphism chips do not contain sufficient numbers of markers in PAR1 and PAR2. The number of markers in PAR1, needed in indirect association studies, should be much larger than for autosomal regions alike in size, since linkage disequilibrium is very low. For genome-wide studies, it is essential to include pseudoautosomal markers since such expensive studies cannot afford to oversee pseudoautosomal linkage or association. This drawback could be solved with a sufficient number of markers, statistical methods that are adopted for the PARs, and their integration into softwares. PMID- 18398440 TI - Ischaemic stroke in hypertensive patients is associated with variations in the PDE4D genome region. AB - Previous Icelandic studies reported that single nucleotide polymorphisms (SNPs) in the phosphodiesterase 4D (PDE4D) region and the 5-lipoxygenase activating protein ALOX5AP were associated with ischaemic stroke, whereas other studies reported ambiguous findings. We examined 932 ischaemic stroke patients from a Swedish population-based stroke register, and 396 control subjects. We assessed possible associations between ischaemic stroke and nine preselected SNPs in the chromosome regions of the PDE4D gene, including rs12188950 (SNP45) and rs3887175 (SNP39); the ALOX5AP gene, including rs17222814 (SG13S25) and the promoter region of the MHC class II transactivator, MHC2TA. The T allele of SNP45 showed negative association with ischaemic stroke (odds ratio, OR=0.72; 95% confidence interval (CI): 0.58-0.91; P=0.0055). Among hypertensive subjects, this influence of the T allele of SNP45, and the T allele of SNP39, were more pronounced (with OR=0.52; 95% CI: 0.37-0.73; P=0.0001 and OR=0.57; 95% CI: 0.41-0.79; P=0.0007, respectively). These SNPs also interacted with hypertension with a relative excess risk due to interaction of -1.66 (P=0.0002) for SNP45 and -1.65 (P=0.0005) for SNP39. The P-values remained significant after correction for multiple testing. Among nonhypertensives, the A allele of SG13S25 indicated increased stroke risk (OR=1.82; 95% CI: 1.21-2.74; P=0.0039; not significant after Bonferroni correction). SNP45 was associated with ischaemic stroke even when controlling for hypertension, diabetes, heart disease and smoking. Our meta analysis of 13 studies (including ours) showed no overall influence of SNP45 on ischaemic stroke. However, the 13 studies may differ because of nonrandom causes, as suggested by the heterogeneity test (P=0.042). This might support previously undetected mechanisms causing fluctuating ischaemic stroke risk. PMID- 18398441 TI - Fragile X syndrome. AB - Fragile X syndrome, an X-linked dominant disorder with reduced penetrance, is associated with intellectual and emotional disabilities ranging from learning problems to mental retardation, and mood instability to autism. It is most often caused by the transcriptional silencing of the FMR1 gene, due to an expansion of a CGG repeat found in the 5'-untranslated region. The FMR1 gene product, FMRP, is a selective RNA-binding protein that negatively regulates local protein synthesis in neuronal dendrites. In its absence, the transcripts normally regulated by FMRP are over translated. The resulting over abundance of certain proteins results in reduced synaptic strength due to AMPA receptor trafficking abnormalities that lead, at least in part, to the fragile X phenotype. PMID- 18398442 TI - A novel genomic disorder: a deletion of the SACS gene leading to spastic ataxia of Charlevoix-Saguenay. AB - We report a Belgian patient with early-onset cerebellar ataxia, progressive spasticity, learning difficulties and moderate perceptive hearing loss. Array Comparative Genomic Hybridisation (aCGH) detected a 1.54 Mb deletion on chromosome 13q12.12. This microdeletion occurred de novo and encompasses the SACS gene. Mutations in SACS are known to cause a recessive condition, similar to the patient's phenotype, called autosomal recessive spastic ataxia of Charlevoix Saguenay (ARSACS). Sequencing of the remaining SACS allele revealed a hemizygous mutation c.10517T>C in exon 9, resulting in an amino-acid substitution (p.F3506S). This is the first patient with ARSACS that carries a de novo chromosomal deletion comprising SACS. We demonstrate the presence of homologous segmental duplications at the breakpoint-containing regions. This suggests non allelic homologous recombination as the mechanism generating this deletion and explains the previous description of copy number variations of this region. This finding confirms the contribution of aCGH to gene identification in autosomal recessive disorders. PMID- 18398444 TI - Pilot study for utilization of dried blood spots for screening of lead, mercury and cadmium in newborns. AB - The exposure of pregnant women and young children to environmental pollutants is an ongoing concern of state and local public health departments. Of primary concern is the exposure to lead in lead-based paints, methyl mercury in contaminated fish and cadmium present at mining sites. The feasibility, utility and methodology of using blood spot cards collected for new born health screening purposes was studied for use in conducting routine state-wide surveillance of blood lead, mercury and cadmium levels in infants. Homogeneity of different lots of blank filter paper was examined. Mass measurements (weights) of filter paper punches were taken across three different lots of filter paper. Statistical analysis of the data was performed using one-way ANOVA, which indicated no significant difference in the means of all three lots, but high variances were noted. The three metals were examined in three different lots of filter papers purchased from the manufacturer. The lots had measurable amounts of cadmium and lead, but not mercury. Lead spike values were observed for roughly about 7% of the blank samples, indicating heterogeneous distribution of this metal. Statistical analysis of the data was also performed using a two-way ANOVA calculation with Tukey's pairwise comparisons. The results found that total mean metal loadings across the three lots were different. The concentration of the metals can be different from each other and the concentration of any one metal can differ across lots. Stability at different concentrations of the heavy metals in blood spotted onto filter paper with time and storage conditions was examined. Results indicate acceptable performance for at least 8.5 months for lead (near CDC's concern level) and for mercury (near NRC's concern level). The filter paper and blood spots were analyzed for metals using an acid extraction, followed by analysis using an inductively coupled plasma mass spectrometer (ICP-MS). Blood spot cards were studied from four different states across the Rocky Mountain region. Internal blank punches adjacent to the blood spot and actual dried spot punches from the same card were analyzed simultaneously. The blank punch indicated the amount of contamination present in the blood spot sample. Statistical analysis of the data was performed using MANOVA followed by calculations for each metal separately. This method was found to be suitable for assessing maternal exposure to lead and mercury using residual newborn screening specimens. Additional research into the applicability for cadmium is needed. Because of the intrinsic problem of contamination from the skin surface of capillary blood samples or other internal or extraneous sources, automatic re analysis of elevated results assures minimal false positives are reported. PMID- 18398445 TI - Intercity transferability of land use regression models for estimating ambient concentrations of nitrogen dioxide. AB - Land use regression (LUR) is a method for predicting the spatial distribution of traffic-related air pollution. To facilitate risk and exposure assessment, and the design of future monitoring networks and sampling campaigns, we sought to determine the extent to which LUR can be used to predict spatial patterns in air pollution in the absence of dedicated measurements. We evaluate the transferability of one LUR model to two other geographically comparable areas with similar climates and pollution types. The source model, developed in 2003 to estimate ambient nitrogen dioxide (NO(2)) concentrations in Vancouver (BC, Canada) was applied to Victoria (BC, Canada) and Seattle (WA, USA). Model estimates were compared with measurements made with Ogawa passive samplers in both cities. As part of this study, 42 locations were sampled in Victoria for a 2 week period in June 2006. Data obtained for Seattle were collected for a different project at 26 locations in March 2005. We used simple linear regression to evaluate the fit of the source model under three scenarios: (1) using the same variables and coefficients as the source model; (2) using the same variables as the source model, but calculating new coefficients for local calibration; and (3) developing site-specific equations with new variables and coefficients. In Scenario 1, we found that the source model had a better fit in Victoria (R(2)=0.51) than in Seattle (R(2)=0.33). Scenario 2 produced improved R(2)-values in both cities (Victoria=0.58, Seattle=0.65), with further improvement achieved under Scenario 3 (Victoria=0.61, Seattle=0.72). Although it is possible to transfer LUR models between geographically similar cities, success may depend on the between-city consistency of the input data. Modest field sampling campaigns for location-specific model calibration can help to produce transfer models that are equally as predictive as their sources. PMID- 18398446 TI - Separation of uncertainty and interindividual variability in human exposure modeling. AB - The NORMTOX model predicts the lifetime-averaged exposure to contaminants through multiple environmental media, that is, food, air, soil, drinking and surface water. The model was developed to test the coherence of Dutch environmental quality objectives (EQOs). A set of EQOs is called coherent if simultaneous exposure to different environmental media that are all polluted up to their respective EQOs does not result in exceeding the acceptable or tolerable daily intake (ADI or TDI). Aim of the present study is to separate the impact of uncertainty and interindividual variability in coherence predictions with the NORMTOX model. The method is illustrated in a case study for chlorfenvinphos, mercury and nitrate. First, ANOVA was used to calculate interindividual variability in input parameters. Second, nested Monte Carlo simulation was used to propagate uncertainty and interindividual variability separately. Lifetime averaged exposure to chlorfenvinphos, mercury and nitrate was modeled for the Dutch population. Output distributions specified the population fraction at risk, due to a particular exposure, and the reliability of this risk. From the case study, it was obtained that at lifelong exposure to all media polluted up to their standard, 100% of the Dutch population exceeds the ADI for chlorfenvinphos, 15% for mercury and 0% for nitrate. Variance in exposure to chlorfenvinphos, mercury and nitrate is mostly caused by interindividual variability instead of true uncertainty. It is concluded that the likelihood that ADIs of chlorfenvinphos and mercury will be exceeded should be further explored. If exceeding is likely, decision makers should focus on identification of high-risk subpopulations, rather than on additional research to obtain more accurate estimates for particular parameters. PMID- 18398448 TI - 29th congress of the European society of cardiology. PMID- 18398447 TI - Extended peptoids: a new class of oligomers based on aromatic building blocks. AB - Peptoids (N-substituted polyglycines) represent a class of bioinspired oligomers that have unique physical and structural properties. Here we report the construction of "extended peptoids" based on aromatic building blocks, in which the N-alkylaminoacetyl group of the peptoid backbone has been replaced by an N alkylaminomethylbenzoyl spacer. Both meta- and para-bromomethylbenzoic acids were synthesized, providing access to a new class of peptoids. Further, inclusion of hydrophilic side chains confers water solubility to these compounds, showing that, like simple peptoids, extended peptoids add an extra dimension to synthetic polyamide oligomers with potential application in a variety of biological contexts. PMID- 18398449 TI - Metagenomic analysis of human diarrhea: viral detection and discovery. AB - Worldwide, approximately 1.8 million children die from diarrhea annually, and millions more suffer multiple episodes of nonfatal diarrhea. On average, in up to 40% of cases, no etiologic agent can be identified. The advent of metagenomic sequencing has enabled systematic and unbiased characterization of microbial populations; thus, metagenomic approaches have the potential to define the spectrum of viruses, including novel viruses, present in stool during episodes of acute diarrhea. The detection of novel or unexpected viruses would then enable investigations to assess whether these agents play a causal role in human diarrhea. In this study, we characterized the eukaryotic viral communities present in diarrhea specimens from 12 children by employing a strategy of "micro mass sequencing" that entails minimal starting sample quantity (<100 mg stool), minimal sample purification, and limited sequencing (384 reads per sample). Using this methodology we detected known enteric viruses as well as multiple sequences from putatively novel viruses with only limited sequence similarity to viruses in GenBank. PMID- 18398450 TI - When the choice is ours: context and agency modulate the neural bases of decision making. AB - The option to choose between several courses of action is often associated with the feeling of being in control. Yet, in certain situations, one may prefer to decline such agency and instead leave the choice to others. In the present functional magnetic resonance imaging (fMRI) study, we provide evidence that the neural processes involved in decision-making are modulated not only by who controls our choice options (agency), but also by whether we have a say in who is in control (context). The fMRI results are noteworthy in that they reveal specific contributions of the anterior frontomedian cortex (viz. BA 10) and the rostral cingulate zone (RCZ) in decision-making processes. The RCZ is engaged when conditions clearly present us with the most choice options. BA 10 is engaged in particular when the choice is completely ours, as well as when it is completely up to others to choose for us which in turn gives rise to an attribution of control to oneself or someone else, respectively. After all, it does not only matter whether we have any options to choose from, but also who decides on that. PMID- 18398451 TI - Genome-wide analysis of abnormal H3K9 acetylation in cloned mice. AB - Somatic nuclear transfer is a cloning technique that shows great promise in the application to regenerative medicine. Although cloned animals are genetically identical to their donor counterparts, abnormalities in phenotype and gene expression are frequently observed. One hypothesis is that the cause of these abnormalities is due to epigenetic aberration. In this report, we focused our analysis on the acetylation of histone H3 at lysine9 (H3K9Ac). Through the use of whole genome tiling arrays and quantitative PCR, we examined this epigenetic event and directly compared and assessed the differences between a cloned mouse (C1) and its parental nuclear donor (D1) counterpart. We identified 4720 regions of chromosomal DNA that showed notable differences in H3K9Ac and report here many genes identified in these hyper- and hypo-acetylated regions. Analysis of a second clone (C2) and its parental donor counterpart (D2) for H3K9Ac showed a high degree of similarity to the C1/D1 pair. This conservation of aberrant acetylation is suggestive of a reproducible epigenetic phenomenon that may lead to the frequent abnormalities observed in cloned mice, such as obesity. Furthermore, we demonstrated Crp which was identified as a hyper-acetylated gene in this study is related to the body mass, suggesting that Crp is a possible candidate of a cause for the abnormal obesity in cloned mice. In this, one of the first reports describing genome-wide epigenetic aberration between parental and nuclear transfer-cloned mammals, we propose that aberrant acetylation of histones (H3K9Ac) flanking promoter regions highly correlates with gene-expression and may itself be an epigenetic change that accounts for variable expression patterns observed in cloned animals. PMID- 18398452 TI - A comparison of phylogenetic network methods using computer simulation. AB - BACKGROUND: We present a series of simulation studies that explore the relative performance of several phylogenetic network approaches (statistical parsimony, split decomposition, union of maximum parsimony trees, neighbor-net, simulated history recombination upper bound, median-joining, reduced median joining and minimum spanning network) compared to standard tree approaches, (neighbor-joining and maximum parsimony) in the presence and absence of recombination. PRINCIPAL FINDINGS: In the absence of recombination, all methods recovered the correct topology and branch lengths nearly all of the time when the substitution rate was low, except for minimum spanning networks, which did considerably worse. At a higher substitution rate, maximum parsimony and union of maximum parsimony trees were the most accurate. With recombination, the ability to infer the correct topology was halved for all methods and no method could accurately estimate branch lengths. CONCLUSIONS: Our results highlight the need for more accurate phylogenetic network methods and the importance of detecting and accounting for recombination in phylogenetic studies. Furthermore, we provide useful information for choosing a network algorithm and a framework in which to evaluate improvements to existing methods and novel algorithms developed in the future. PMID- 18398453 TI - Learning and innovative elements of strategy adoption rules expand cooperative network topologies. AB - Cooperation plays a key role in the evolution of complex systems. However, the level of cooperation extensively varies with the topology of agent networks in the widely used models of repeated games. Here we show that cooperation remains rather stable by applying the reinforcement learning strategy adoption rule, Q learning on a variety of random, regular, small-word, scale-free and modular network models in repeated, multi-agent Prisoner's Dilemma and Hawk-Dove games. Furthermore, we found that using the above model systems other long-term learning strategy adoption rules also promote cooperation, while introducing a low level of noise (as a model of innovation) to the strategy adoption rules makes the level of cooperation less dependent on the actual network topology. Our results demonstrate that long-term learning and random elements in the strategy adoption rules, when acting together, extend the range of network topologies enabling the development of cooperation at a wider range of costs and temptations. These results suggest that a balanced duo of learning and innovation may help to preserve cooperation during the re-organization of real-world networks, and may play a prominent role in the evolution of self-organizing, complex systems. PMID- 18398454 TI - Acetylation regulates WRN catalytic activities and affects base excision DNA repair. AB - BACKGROUND: The Werner protein (WRN), defective in the premature aging disorder Werner syndrome, participates in a number of DNA metabolic processes, and we have been interested in the possible regulation of its function in DNA repair by post translational modifications. Acetylation mediated by histone acetyltransferases is of key interest because of its potential importance in aging, DNA repair and transcription. METHODOLOGY/PRINCIPAL FINDINGS: Here, we have investigated the p300 acetylation mediated changes on the function of WRN in base excision DNA repair (BER). We show that acetylation of WRN increases in cells treated with methyl methanesulfonate (MMS), suggesting that acetylation of WRN may play a role in response to DNA damage. This hypothesis is consistent with our findings that acetylation of WRN stimulates its catalytic activities in vitro and in vivo, and that acetylated WRN enhances pol beta-mediated strand displacement DNA synthesis more than unacetylated WRN. Furthermore, we show that cellular exposure to the histone deacetylase inhibitor sodium butyrate stimulates long patch BER in wild type cells but not in WRN depleted cells, suggesting that acetylated WRN participates significantly in this process. CONCLUSION/SIGNIFICANCE: Collectively, these results provide the first evidence for a specific role of p300 mediated WRN acetylation in regulating its function during BER. PMID- 18398455 TI - Mapping genetically compensatory pathways from synthetic lethal interactions in yeast. AB - BACKGROUND: Synthetic lethal genetic interaction analysis has been successfully applied to predicting the functions of genes and their pathway identities. In the context of synthetic lethal interaction data alone, the global similarity of synthetic lethal interaction patterns between two genes is used to predict gene function. With physical interaction data, such as protein-protein interactions, the enrichment of physical interactions within subsets of genes and the enrichment of synthetic lethal interactions between those subsets of genes are used as an indication of compensatory pathways. RESULT: In this paper, we propose a method of mapping genetically compensatory pathways from synthetic lethal interactions. Our method is designed to discover pairs of gene-sets in which synthetic lethal interactions are depleted among the genes in an individual set and where such gene-set pairs are connected by many synthetic lethal interactions. By its nature, our method could select compensatory pathway pairs that buffer the deleterious effect of the failure of either one, without the need of physical interaction data. By focusing on compensatory pathway pairs where genes in each individual pathway have a highly homogenous cellular function, we show that many cellular functions have genetically compensatory properties. CONCLUSION: We conclude that synthetic lethal interaction data are a powerful source to map genetically compensatory pathways, especially in systems lacking physical interaction information, and that the cellular function network contains abundant compensatory properties. PMID- 18398456 TI - Electronically switchable sham transcranial magnetic stimulation (TMS) system. AB - Transcranial magnetic stimulation (TMS) is increasingly being used to demonstrate the causal links between brain and behavior in humans. Further, extensive clinical trials are being conducted to investigate the therapeutic role of TMS in disorders such as depression. Because TMS causes strong peripheral effects such as auditory clicks and muscle twitches, experimental artifacts such as subject bias and placebo effect are clear concerns. Several sham TMS methods have been developed, but none of the techniques allows one to intermix real and sham TMS on a trial-by-trial basis in a double-blind manner. We have developed an attachment that allows fast, automated switching between Standard TMS and two types of control TMS (Sham and Reverse) without movement of the coil or reconfiguration of the setup. We validate the setup by performing mathematical modeling, search-coil and physiological measurements. To see if the stimulus conditions can be blinded, we conduct perceptual discrimination and sensory perception studies. We verify that the physical properties of the stimulus are appropriate, and that successive stimuli do not contaminate each other. We find that the threshold for motor activation is significantly higher for Reversed than for Standard stimulation, and that Sham stimulation entirely fails to activate muscle potentials. Subjects and experimenters perform poorly at discriminating between Sham and Standard TMS with a figure-of-eight coil, and between Reverse and Standard TMS with a circular coil. Our results raise the possibility of utilizing this technique for a wide range of applications. PMID- 18398457 TI - Oral high-dose atorvastatin treatment in relapsing-remitting multiple sclerosis. AB - BACKGROUND: Recent data from animal models of multiple sclerosis (MS) and from a pilot study indicated a possible beneficial impact of statins on MS. METHODOLOGY/PRINCIPAL FINDINGS: Safety, tolerability and effects on disease activity of atorvastatin given alone or in combination with interferon-beta (IFN beta) were assessed in a phase II open-label baseline-to-treatment trial in relapsing-remitting MS (RRMS). Patients with at least one gadolinium-enhancing lesion (CEL) at screening by magnetic resonance imaging (MRI) were eligible for the study. After a baseline period of 3 monthly MRI scans (months -2 to 0), patients followed a 9-month treatment period on 80 mg atorvastatin daily. The number of CEL in treatment months 6 to 9 compared to baseline served as the primary endpoint. Other MRI-based parameters as well as changes in clinical scores and immune responses served as secondary endpoints. Of 80 RRMS patients screened, 41 were included, among them 16 with IFN-beta comedication. The high dose of 80 mg atorvastatin was well tolerated in the majority of patients, regardless of IFN-beta comedication. Atorvastatin treatment led to a substantial reduction in the number and volume of CEL in two-sided multivariate analysis (p = 0.003 and p = 0.008). A trend towards a significant decrease in number and volume of CEL was also detected in patients with IFN-beta comedication (p = 0.060 and p = 0.062), in contrast to patients without IFN-beta comedication (p = 0.170 and p = 0.140). Immunological investigations showed no suppression in T cell response but a significant increase in IL-10 production. CONCLUSIONS/SIGNIFICANCE: Our data suggest that high-dose atorvastatin treatment in RRMS is safe and well tolerated. Moreover, MRI analysis indicates a possible beneficial effect of atorvastatin, alone or in combination with IFN-beta, on the development of new CEL. Thus, our findings provide a rationale for phase II/III trials, including combination of atorvastatin with already approved immunomodulatory therapy regimens. TRIAL REGISTRATION: ClinicalTrials.gov NCT00616187. PMID- 18398458 TI - How to derive biological information from the value of the normalization constant in allometric equations. AB - Allometric equations are widely used in many branches of biological science. The potential information content of the normalization constant b in allometric equations of the form Y = bX(a) has, however, remained largely neglected. To demonstrate the potential for utilizing this information, I generated a large number of artificial datasets that resembled those that are frequently encountered in biological studies, i.e., relatively small samples including measurement error or uncontrolled variation. The value of X was allowed to vary randomly within the limits describing different data ranges, and a was set to a fixed theoretical value. The constant b was set to a range of values describing the effect of a continuous environmental variable. In addition, a normally distributed random error was added to the values of both X and Y. Two different approaches were then used to model the data. The traditional approach estimated both a and b using a regression model, whereas an alternative approach set the exponent a at its theoretical value and only estimated the value of b. Both approaches produced virtually the same model fit with less than 0.3% difference in the coefficient of determination. Only the alternative approach was able to precisely reproduce the effect of the environmental variable, which was largely lost among noise variation when using the traditional approach. The results show how the value of b can be used as a source of valuable biological information if an appropriate regression model is selected. PMID- 18398459 TI - The effect of diagnostic delays on the drop-out rate and the total delay to diagnosis of tuberculosis. AB - BACKGROUND: Numerous patient and healthcare system-related delays contribute to the overall delay experienced by patients from onset of TB symptoms to diagnosis and treatment. Such delays are critical as infected individuals remain untreated in the community, providing more opportunities for transmission of the disease and adversely affecting the epidemic. METHODOLOGY/PRINCIPAL FINDINGS: We present an analysis of the factors that contribute to the overall delay in TB diagnosis and treatment, in a resource-poor setting. Impact on the distribution of diagnostic delay times was assessed for various factors, the sensitivity of the diagnostic method being found to be the most significant. A linear relationship was found between the sensitivity of the test and the predicted mean delay time, with an increase in test sensitivity resulting in a reduced mean delay time and a reduction in the drop-out rate. CONCLUSIONS/SIGNIFICANCE: The results show that in a developing country a number of delay factors, particularly the low sensitivity of the initial sputum smear microscopy test, potentially increase total diagnostic delay times experienced by TB patients significantly. The results reinforce the urgent need for novel diagnostic methods, both for smear positive and negative TB, that are highly sensitive, accessible and point of care, in order to reduce mean delay times. PMID- 18398460 TI - A randomized placebo-controlled trial of intermittent preventive treatment in pregnant women in the context of insecticide treated nets delivered through the antenatal clinic. AB - BACKGROUND: Current recommendations to prevent malaria in African pregnant women rely on insecticide treated nets (ITNs) and intermittent preventive treatment (IPTp). However, there is no information on the safety and efficacy of their combined use. METHODS: 1030 pregnant Mozambican women of all gravidities received a long-lasting ITN during antenatal clinic (ANC) visits and, irrespective of HIV status, were enrolled in a randomised, double blind, placebo-controlled trial, to assess the safety and efficacy of 2-dose sulphadoxine-pyrimethamine (SP). The main outcome was the reduction in low birth weight. FINDINGS: Two-dose SP was safe and well tolerated, but was not associated with reductions in anaemia prevalence at delivery (RR, 0.92 [95% CI, 0.79-1.08]), low birth weight (RR, 0.99 [95% CI, 0.70-1.39]), or overall placental infection (p = 0.964). However, the SP group showed a 40% reduction (95% CI, 7.40-61.20]; p = 0.020) in the incidence of clinical malaria during pregnancy, and reductions in the prevalence of peripheral parasitaemia (7.10% vs 15.15%) (p<0.001), and of actively infected placentas (7.04% vs 13.60%) (p = 0.002). There was a reduction in severe anaemia at delivery of borderline statistical significance (p = 0.055). These effects were not modified by gravidity or HIV status. Reported ITN's use was more than 90% in both groups. CONCLUSIONS: Two-dose SP was associated with a reduction in some indicators, but these were not translated to significant improvement in other maternal or birth outcomes. The use of ITNs during pregnancy may reduce the need to administer IPTp. ITNs should be part of the ANC package in sub-Saharan Africa. TRIAL REGISTRATION: ClinicalTrials.gov NCT00209781. PMID- 18398461 TI - S1 constrains S4 in the voltage sensor domain of Kv7.1 K+ channels. AB - Voltage-gated K(+) channels comprise a central pore enclosed by four voltage sensing domains (VSDs). While movement of the S4 helix is known to couple to channel gate opening and closing, the nature of S4 motion is unclear. Here, we substituted S4 residues of Kv7.1 channels by cysteine and recorded whole-cell mutant channel currents in Xenopus oocytes using the two-electrode voltage-clamp technique. In the closed state, disulfide and metal bridges constrain residue S225 (S4) nearby C136 (S1) within the same VSD. In the open state, two neighboring I227 (S4) are constrained at proximity while residue R228 (S4) is confined close to C136 (S1) of an adjacent VSD. Structural modeling predicts that in the closed to open transition, an axial rotation (approximately 190 degrees) and outward translation of S4 (approximately 12 A) is accompanied by VSD rocking. This large sensor motion changes the intra-VSD S1-S4 interaction to an inter-VSD S1-S4 interaction. These constraints provide a ground for cooperative subunit interactions and suggest a key role of the S1 segment in steering S4 motion during Kv7.1 gating. PMID- 18398462 TI - Glial progenitor-like phenotype in low-grade glioma and enhanced CD133-expression and neuronal lineage differentiation potential in high-grade glioma. AB - BACKGROUND: While neurosphere- as well as xenograft tumor-initiating cells have been identified in gliomas, the resemblance between glioma cells and neural stem/progenitor cells as well as the prognostic value of stem/progenitor cell marker expression in glioma are poorly clarified. METHODOLOGY/PRINCIPAL FINDINGS: Viable glioma cells were characterized for surface marker expression along the glial genesis hierarchy. Six low-grade and 17 high-grade glioma specimens were flow-cytometrically analyzed for markers characteristics of stem cells (CD133); glial progenitors (PDGFRalpha, A2B5, O4, and CD44); and late oligodendrocyte progenitors (O1). In parallel, the expression of glial fibrillary acidic protein (GFAP), synaptophysin and neuron-specific enolase (NSE) was immunohistochemically analyzed in fixed tissue specimens. Irrespective of the grade and morphological diagnosis of gliomas, glioma cells concomitantly expressed PDGFRalpha, A2B5, O4, CD44 and GFAP. In contrast, O1 was weakly expressed in all low-grade and the majority of high-grade glioma specimens analyzed. Co-expression of neuronal markers was observed in all high-grade, but not low-grade, glioma specimens analyzed. The rare CD133 expressing cells in low-grade glioma specimens typically co-expressed vessel endothelial marker CD31. In contrast, distinct CD133 expression profiles in up to 90% of CD45-negative glioma cells were observed in 12 of the 17 high-grade glioma specimens and the majority of these CD133 expressing cells were CD31 negative. The CD133 expression correlates inversely with length of patient survival. Surprisingly, cytogenetic analysis showed that gliomas contained normal and abnormal cell karyotypes with hitherto indistinguishable phenotype. CONCLUSIONS/SIGNIFICANCE: This study constitutes an important step towards clarification of lineage commitment and differentiation blockage of glioma cells. Our data suggest that glioma cells may resemble expansion of glial lineage progenitor cells with compromised differentiation capacity downstream of A2B5 and O4 expression. The concurrent expression of neuronal markers demonstrates that high-grade glioma cells are endowed with multi lineage differentiation potential in vivo. Importantly, enhanced CD133 expression marks a poor prognosis in gliomas. PMID- 18398463 TI - Environmental adaptation: genomic analysis of the piezotolerant and psychrotolerant deep-sea iron reducing bacterium Shewanella piezotolerans WP3. AB - Shewanella species are widespread in various environments. Here, the genome sequence of Shewanella piezotolerans WP3, a piezotolerant and psychrotolerant iron reducing bacterium from deep-sea sediment was determined with related functional analysis to study its environmental adaptation mechanisms. The genome of WP3 consists of 5,396,476 base pairs (bp) with 4,944 open reading frames (ORFs). It possesses numerous genes or gene clusters which help it to cope with extreme living conditions such as genes for two sets of flagellum systems, structural RNA modification, eicosapentaenoic acid (EPA) biosynthesis and osmolyte transport and synthesis. And WP3 contains 55 open reading frames encoding putative c-type cytochromes which are substantial to its wide environmental adaptation ability. The mtr-omc gene cluster involved in the insoluble metal reduction in the Shewanella genus was identified and compared. The two sets of flagellum systems were found to be differentially regulated under low temperature and high pressure; the lateral flagellum system was found essential for its motility and living at low temperature. PMID- 18398464 TI - The homeodomain derived peptide Penetratin induces curvature of fluid membrane domains. AB - BACKGROUND: Protein membrane transduction domains that are able to cross the plasma membrane are present in several transcription factors, such as the homeodomain proteins and the viral proteins such as Tat of HIV-1. Their discovery resulted in both new concepts on the cell communication during development, and the conception of cell penetrating peptide vectors for internalisation of active molecules into cells. A promising cell penetrating peptide is Penetratin, which crosses the cell membranes by a receptor and metabolic energy-independent mechanism. Recent works have claimed that Penetratin and similar peptides are internalized by endocytosis, but other endocytosis-independent mechanisms have been proposed. Endosomes or plasma membranes crossing mechanisms are not well understood. Previously, we have shown that basic peptides induce membrane invaginations suggesting a new mechanism for uptake, "physical endocytosis". METHODOLOGY/PRINCIPAL FINDINGS: Herein, we investigate the role of membrane lipid phases on Penetratin induced membrane deformations (liquid ordered such as in "raft" microdomains versus disordered fluid "non-raft" domains) in membrane models. Experimental data show that zwitterionic lipid headgroups take part in the interaction with Penetratin suggesting that the external leaflet lipids of cells plasma membrane are competent for peptide interaction in the absence of net negative charges. NMR and X-ray diffraction data show that the membrane perturbations (tubulation and vesiculation) are associated with an increase in membrane negative curvature. These effects on curvature were observed in the liquid disordered but not in the liquid ordered (raft-like) membrane domains. CONCLUSIONS/SIGNIFICANCE: The better understanding of the internalisation mechanisms of protein transduction domains will help both the understanding of the mechanisms of cell communication and the development of potential therapeutic molecular vectors. Here we showed that the membrane targets for these molecules are preferentially the fluid membrane domains and that the mechanism involves the induction of membrane negative curvature. Consequences on cellular uptake are discussed. PMID- 18398465 TI - Reconstruction of cell lineage trees in mice. AB - The cell lineage tree of a multicellular organism represents its history of cell divisions from the very first cell, the zygote. A new method for high-resolution reconstruction of parts of such cell lineage trees was recently developed based on phylogenetic analysis of somatic mutations accumulated during normal development of an organism. In this study we apply this method in mice to reconstruct the lineage trees of distinct cell types. We address for the first time basic questions in developmental biology of higher organisms, namely what is the correlation between the lineage relation among cells and their (1) function, (2) physical proximity and (3) anatomical proximity. We analyzed B-cells, kidney , mesenchymal- and hematopoietic-stem cells, as well as satellite cells, which are adult skeletal muscle stem cells isolated from their niche on the muscle fibers (myofibers) from various skeletal muscles. Our results demonstrate that all analyzed cell types are intermingled in the lineage tree, indicating that none of these cell types are single exclusive clones. We also show a significant correlation between the physical proximity of satellite cells within muscles and their lineage. Furthermore, we show that satellite cells obtained from a single myofiber are significantly clustered in the lineage tree, reflecting their common developmental origin. Lineage analysis based on somatic mutations enables performing high resolution reconstruction of lineage trees in mice and humans, which can provide fundamental insights to many aspects of their development and tissue maintenance. PMID- 18398467 TI - Measles on the edge: coastal heterogeneities and infection dynamics. AB - Mathematical models can help elucidate the spatio-temporal dynamics of epidemics as well as the impact of control measures. The gravity model for directly transmitted diseases is currently one of the most parsimonious models for spatial epidemic spread. This model uses distance-weighted, population size-dependent coupling to estimate host movement and disease incidence in metapopulations. The model captures overall measles dynamics in terms of underlying human movement in pre-vaccination England and Wales (previously established). In spatial models, edges often present a special challenge. Therefore, to test the model's robustness, we analyzed gravity model incidence predictions for coastal cities in England and Wales. Results show that, although predictions are accurate for inland towns, they significantly underestimate coastal persistence. We examine incidence, outbreak seasonality, and public transportation records, to show that the model's inaccuracies stem from an underestimation of total contacts per individual along the coast. We rescue this predicted 'edge effect' by increasing coastal contacts to approximate the number of per capita inland contacts. These results illustrate the impact of 'edge effects' on epidemic metapopulations in general and illustrate directions for the refinement of spatiotemporal epidemic models. PMID- 18398466 TI - Vitellogenin functions as a multivalent pattern recognition receptor with an opsonic activity. AB - BACKGROUND: Vitellogenin (Vg), a major reproductive protein, has been associated with infection-resistant response in fish. However, the underlying mechanisms by which Vg is involved in anti-infectious response are not understood. METHODOLOGY/RESULTS: By both protein-microbe interaction analysis and enzyme linked immunosorbent assay as well as phagocytosis test, we demonstrate for the first time that fish Vg acts as a pattern recognition molecule with multiple specificities that can recognize bacteria as well as fungus rather than self components from fish, and functions as an opsonin that can enhance macrophage phagocytosis. CONCLUSIONS: This study shows that fish Vg plays an integrative function in regulating immunity via its pleiotropic effects on both recognizing pathogen-associated molecular patterns and promoting macrophage phagocytosis. It also supports the notion that factors normally involved in control of female reproduction are associated with immunity in organisms that rely on Vg for oocyte development. PMID- 18398468 TI - The Cayman crab fly revisited--phylogeny and biology of Drosophila endobranchia. AB - BACKGROUND: The majority of all known drosophilid flies feed on microbes. The wide spread of microorganisms consequently mean that drosophilids also can be found on a broad range of substrates. One of the more peculiar types of habitat is shown by three species of flies that have colonized land crabs. In spite of their intriguing lifestyle, the crab flies have remained poorly studied. Perhaps the least investigated of the three crab flies is the Cayman Island endemic Drosophila endobranchia. Apart from its life cycle very little is known about this species, including its phylogenetic position, which has remained unresolved due to a cryptic set of characteristics. PRINCIPAL FINDINGS: Based on molecular data, corroborated by a re-analysis of the morphological make up, we have resolved the phylogenetic position of D. endobranchia and show that it somewhat surprisingly belongs to the large Neotropical repleta radiation, and should be considered as an aberrant member of the canalinea species group. Furthermore we also provide additional data on the behavior of these remarkable flies. CONCLUSION: Our findings reveal that the two Caribbean crab flies are not as distantly related as first thought, as both species are members of the derived repleta radiation. That this lineage has given rise to two species with the same odd type of breeding substrate is curious and prompts the question of what aspects of their shared ancestry has made these flies suitable for a life on (and inside) land crabs. Knowledge of the phylogenetic position of D. endobranchia will allow for comparative explorations and will aid in efforts aimed at understanding processes involved in drastic host shifts and extreme specialization. PMID- 18398469 TI - KCNE1 constrains the voltage sensor of Kv7.1 K+ channels. AB - Kv7 potassium channels whose mutations cause cardiovascular and neurological disorders are members of the superfamily of voltage-gated K(+) channels, comprising a central pore enclosed by four voltage-sensing domains (VSDs) and sharing a homologous S4 sensor sequence. The Kv7.1 pore-forming subunit can interact with various KCNE auxiliary subunits to form K(+) channels with very different gating behaviors. In an attempt to characterize the nature of the promiscuous gating of Kv7.1 channels, we performed a tryptophan-scanning mutagenesis of the S4 sensor and analyzed the mutation-induced perturbations in gating free energy. Perturbing the gating energetics of Kv7.1 bias most of the mutant channels towards the closed state, while fewer mutations stabilize the open state or the inactivated state. In the absence of auxiliary subunits, mutations of specific S4 residues mimic the gating phenotypes produced by co assembly of Kv7.1 with either KCNE1 or KCNE3. Many S4 perturbations compromise the ability of KCNE1 to properly regulate Kv7.1 channel gating. The tryptophan induced packing perturbations and cysteine engineering studies in S4 suggest that KCNE1 lodges at the inter-VSD S4-S1 interface between two adjacent subunits, a strategic location to exert its striking action on Kv7.1 gating functions. PMID- 18398470 TI - Negative correlation between brain glutathione level and negative symptoms in schizophrenia: a 3T 1H-MRS study. AB - BACKGROUND: Glutathione (GSH), a major intracellular antioxidant, plays a role in NMDA receptor-mediated neurotransmission, which is involved in the pathophysiology of schizophrenia. In the present study, we aimed to investigate whether GSH levels are altered in the posterior medial frontal cortex of schizophrenic patients. Furthermore, we examined correlations between GSH levels and clinical variables in patients. METHODS AND FINDINGS: Twenty schizophrenia patients and 16 age- and gender-matched normal controls were enrolled to examine the levels of GSH in the posterior medial frontal cortex by using 3T SIGNA EXCITE (1)H-MRS with the spectral editing technique, MEGA-PRESS. Clinical variables of patients were assessed by the Global Assessment of Functioning (GAF), Scale for the Assessment of Negative Symptoms (SANS), Brief Psychiatric Rating Scale (BPRS), Drug-Induced Extra-Pyramidal Symptoms Scale (DIEPSS), and five cognitive performance tests (Word Fluency Test, Stroop Test, Trail Making Test, Wisconsin Card Sorting Test and Digit Span Distractibility Test). Levels of GSH in the posterior medial frontal cortex of schizophrenic patients were not different from those of normal controls. However, we found a significant negative correlation between GSH levels and the severity of negative symptoms (SANS total score and negative symptom subscore on BPRS) in patients. There were no correlations between brain GSH levels and scores on any cognitive performance test except Trail Making Test part A. CONCLUSION: These results suggest that GSH levels in the posterior medial frontal cortex may be related to negative symptoms in schizophrenic patients. Therefore, agents that increase GSH levels in the brain could be potential therapeutic drugs for negative symptoms in schizophrenia. PMID- 18398471 TI - Identification of a novel retinoid by small molecule screening with zebrafish embryos. AB - Small molecules have played an important role in delineating molecular pathways involved in embryonic development and disease pathology. The need for novel small molecule modulators of biological processes has driven a number of targeted screens on large diverse libraries. However, due to the specific focus of such screens, the majority of the bioactive potential of these libraries remains unharnessed. In order to identify a higher proportion of compounds with interesting biological activities, we screened a diverse synthetic library for compounds that perturb the development of any of the multiple organs in zebrafish embryos. We identified small molecules that affect the development of a variety of structures such as heart, vasculature, brain, and body-axis. We utilized the previously known role of retinoic acid in anterior-posterior (A-P) patterning to identify the target of DTAB, a compound that caused A-P axis shortening in the zebrafish embryo. We show that DTAB is a retinoid with selective activity towards retinoic acid receptors gamma and beta. Thus, conducting zebrafish developmental screens using small molecules will not only enable the identification of compounds with diverse biological activities in a large chemical library but may also facilitate the identification of the target pathways of these biologically active molecules. PMID- 18398472 TI - Association of common polymorphisms in GLUT9 gene with gout but not with coronary artery disease in a large case-control study. AB - BACKGROUND: Serum uric acid (UA) levels have recently been shown to be genetically influenced by common polymorphisms in the GLUT9 gene in two genome wide association analyses of Italian and British populations. Elevated serum UA levels are often found in conjunction with the metabolic syndrome. Hyperuricemia is the major risk factor for gout and has been associated with increased cardiovascular morbidity and mortality. The aim of the present study was to further elucidate the association of polymorphisms in GLUT9 with gout and coronary artery disease (CAD) or myocardial infarction (MI). To test our hypotheses, we performed two large case-control association analyses of individuals from the German MI Family Study. METHODS AND FINDINGS: First, 665 patients with gout and 665 healthy controls, which were carefully matched for age and gender, were genotyped for four single nucleotide polymorphisms (SNPs) within or near the GLUT9 gene. All four SNPs demonstrated highly significant association with gout. SNP rs6855911, located within intron 7 of GLUT9, showed the strongest signal with a protective effect of the minor allele with an allelic odds ratio of 0.62 (95% confidence interval 0.52-0.75; p = 3.2*10(-7)). Importantly, this finding was not influenced by adjustment for components of the metabolic syndrome or intake of diuretics. Secondly, 1,473 cases with severe CAD or MI and 1,241 healthy controls were tested for the same four GLUT9 SNPs. The analyses revealed, however, no significant association with CAD or with MI. Additional screening of genome-wide association data sets showed no signal for CAD or MI within the GLUT9 gene region. CONCLUSION: Thus, our results provide compelling evidence that common genetic variations within the GLUT9 gene strongly influence the risk for gout but are unlikely to have a major effect on CAD or MI in a German population. PMID- 18398473 TI - Late entry to HIV care limits the impact of anti-retroviral therapy in The Netherlands. AB - OBJECTIVE: To explain differences in survival in the first three years of combination anti-retroviral therapy (cART) between HIV treatment centres in The Netherlands. METHODOLOGY/PRINCIPAL FINDINGS: We developed a mathematical simulation model, parameterised using data from the ATHENA cohort that describes patients entering care, being monitored and starting cART. Three scenarios were used to represent three treatment centres with widely varying mortality rates on cART that were differentiated by: (i) the distribution of CD4 counts of patients entering care; (ii) the age distribution of patients entering care; (iii) the average rate of monitoring the patients not on cART. At the level of the treatment centre, the fraction of Dutch MSM dying in the first three years of treatment ranged from 0% to 8%. The mathematical model captured the large variation in observed mortality between the three treatment centres. Manipulating the age-distribution of patients or the frequency of monitoring did not affect the model predictions. In contrast, when the same national average distribution of CD4 count at entry was used in all the scenarios, the variation in predicted mortality between all centres was diminished. CONCLUSIONS/SIGNIFICANCE: Patients entering care with low CD4 counts appears to be the main source of variation in the mortality rates between Dutch treatment centres. Recruiting HIV-infected individuals to care earlier could lead to substantial improvements in cART outcomes. For example, if patients were to present with at least 400 CD4 cells/mm(3), as they do already in some centres, then our model predicts that the mortality in the first three years of cART could be reduced by approximately 20%. PMID- 18398474 TI - An information-theoretic analysis of genetics, gender and age in cancer patients. AB - Germline genetics, gender and hormonal-signaling pathways are all well described modifiers of cancer risk and progression. Although an improved understanding of how germline genetic variants interact with other cancer risk factors may allow better prevention and treatment of human cancer, measuring and quantifying these interactions is challenging. In other areas of research, Information Theory has been used to quantitatively describe similar multivariate interactions. We implemented a novel information-theoretic analysis to measure the joint effect of a high frequency germline genetic variant of the p53 tumor suppressor pathway (MDM2 SNP309 T/G) and gender on clinical cancer phenotypes. This analysis quantitatively describes synergistic interactions among gender, the MDM2 SNP309 locus, and the age of onset of tumorigenesis in p53 mutation carriers. These results offer a molecular and genetic basis for the observed sexual dimorphism of cancer risk in p53 mutation carriers and a model is proposed that suggests a novel cancer prevention strategy for p53 mutation carriers. PMID- 18398475 TI - Safety and immunogenicity of a malaria vaccine, Plasmodium falciparum AMA-1/MSP-1 chimeric protein formulated in montanide ISA 720 in healthy adults. AB - BACKGROUND: The P. falciparum chimeric protein 2.9 (PfCP-2.9) consisting of the sequences of MSP1-19 and AMA-1 (III) is a malaria vaccine candidate that was found to induce inhibitory antibodies in rabbits and monkeys. This was a phase I randomized, single-blind, placebo-controlled, dose-escalation study to evaluate the safety and immunogenicity of the PfCP-2.9 formulated with a novel adjuvant Montanide ISA720. Fifty-two subjects were randomly assigned to 4 dose groups of 10 participants, each receiving the test vaccine of 20, 50, 100, or 200 microg respectively, and 1 placebo group of 12 participants receiving the adjuvant only. METHODS AND FINDINGS: The vaccine formulation was shown to be safe and well tolerated, and none of the participants withdrew. The total incidence of local adverse events (AEs) was 75%, distributed among 58% of the placebo group and 80% of those vaccinated. Among the vaccinated, 65% had events that were mild and 15% experienced moderate AEs. Almost all systemic adverse reactions observed in this study were graded as mild and required no therapy. The participants receiving the test vaccine developed detectable antibody responses which were boosted by the repeated vaccinations. Sixty percent of the vaccinated participants had high ELISA titers (>1:10,000) of antigen-specific antibodies which could also recognize native parasite proteins in an immunofluorescence assay (IFA). CONCLUSION: This study is the first clinical trial for this candidate and builds on previous investigations supporting PfCP-2.9/ISA720 as a promising blood-stage malaria vaccine. Results demonstrate safety, tolerability (particularly at the lower doses tested) and immunogenicity of the formulation. Further clinical development is ongoing to explore optimizing the dose and schedule of the formulation to decrease reactogenicity without compromising immunogenicity. TRIAL REGISTRATION: Chinese State Food and Drug Administration (SFDA) 2002SL0046; Controlled-Trials.com ISRCTN66850051 [66850051]. PMID- 18398476 TI - Sequence effect of self-assembling peptides on the complexation and in vitro delivery of the hydrophobic anticancer drug ellipticine. AB - A special class of self-assembling peptides has been found to be capable of stabilizing the hydrophobic anticancer agent ellipticine in aqueous solution. Here we study the effect of peptide sequence on the complex formation and its anticancer activity in vitro. Three peptides, EAK16-II, EAK16-IV and EFK16-II, were selected to have either a different charge distribution (EAK16-II vs. EAK16 IV) or a varying hydrophobicity (EAK16-II vs. EFK16-II). Results on their complexation with ellipticine revealed that EAK16-II and EAK16-IV were able to stabilize protonated ellipticine or ellipticine microcrystals depending on the peptide concentration; EFK16-II could stabilize neutral ellipticine molecules and ellipticine microcrystals. These different molecular states of ellipticine were expected to affect ellipticine delivery. The anticancer activity of these complexes was tested against two cancer cell lines: A549 and MCF-7, and related to the cell viability. The viability results showed that the complexes with protonated ellipticine were effective in eradicating both cancer cells (viability <0.05), but their dilutions in water were not stable, leading to a fast decrease in their toxicity. In contrast, the complexes formulated with EFK16-II were relatively stable upon dilution, but their original toxicity was relatively low compared to that with protonated ellipticine. Overall, the charge distribution of the peptides seemed not to affect the complex formation and its therapeutic efficacy in vitro; however, the increase in hydrophobicity of the peptides significantly altered the state of stabilized ellipticine and increased the stability of the complexes. This work provides essential information for peptide sequence design in the development of self-assembling peptide-based delivery of hydrophobic anticancer drugs. PMID- 18398477 TI - Local translation in primary afferent fibers regulates nociception. AB - Recent studies have demonstrated the importance of local protein synthesis for neuronal plasticity. In particular, local mRNA translation through the mammalian target of rapamycin (mTOR) has been shown to play a key role in regulating dendrite excitability and modulating long-term synaptic plasticity associated with learning and memory. There is also increased evidence to suggest that intact adult mammalian axons have a functional requirement for local protein synthesis in vivo. Here we show that the translational machinery is present in some myelinated sensory fibers and that active mTOR-dependent pathways participate in maintaining the sensitivity of a subpopulation of fast-conducting nociceptors in vivo. Phosphorylated mTOR together with other downstream components of the translational machinery were localized to a subset of myelinated sensory fibers in rat cutaneous tissue. We then showed with electromyographic studies that the mTOR inhibitor rapamycin reduced the sensitivity of a population of myelinated nociceptors known to be important for the increased mechanical sensitivity that follows injury. Behavioural studies confirmed that local treatment with rapamycin significantly attenuated persistent pain that follows tissue injury, but not acute pain. Specifically, we found that rapamycin blunted the heightened response to mechanical stimulation that develops around a site of injury and reduced the long-term mechanical hypersensitivity that follows partial peripheral nerve damage--a widely used model of chronic pain. Our results show that the sensitivity of a subset of sensory fibers is maintained by ongoing mTOR-mediated local protein synthesis and uncover a novel target for the control of long-term pain states. PMID- 18398478 TI - Onset dynamics of action potentials in rat neocortical neurons and identified snail neurons: quantification of the difference. AB - The generation of action potentials (APs) is a key process in the operation of nerve cells and the communication between neurons. Action potentials in mammalian central neurons are characterized by an exceptionally fast onset dynamics, which differs from the typically slow and gradual onset dynamics seen in identified snail neurons. Here we describe a novel method of analysis which provides a quantitative measure of the onset dynamics of action potentials. This method captures the difference between the fast, step-like onset of APs in rat neocortical neurons and the gradual, exponential-like AP onset in identified snail neurons. The quantitative measure of the AP onset dynamics, provided by the method, allows us to perform quantitative analyses of factors influencing the dynamics. PMID- 18398479 TI - Nature of protein family signatures: insights from singular value analysis of position-specific scoring matrices. AB - Position-specific scoring matrices (PSSMs) are useful for detecting weak homology in protein sequence analysis, and they are thought to contain some essential signatures of the protein families. In order to elucidate what kind of ingredients constitute such family-specific signatures, we apply singular value decomposition to a set of PSSMs and examine the properties of dominant right and left singular vectors. The first right singular vectors were correlated with various amino acid indices including relative mutability, amino acid composition in protein interior, hydropathy, or turn propensity, depending on proteins. A significant correlation between the first left singular vector and a measure of site conservation was observed. It is shown that the contribution of the first singular component to the PSSMs act to disfavor potentially but falsely functionally important residues at conserved sites. The second right singular vectors were highly correlated with hydrophobicity scales, and the corresponding left singular vectors with contact numbers of protein structures. It is suggested that sequence alignment with a PSSM is essentially equivalent to threading supplemented with functional information. In addition, singular vectors may be useful for analyzing and annotating the characteristics of conserved sites in protein families. PMID- 18398480 TI - COPI vesicle transport is a common requirement for tube expansion in Drosophila. AB - BACKGROUND: Tube expansion defects like stenoses and atresias cause devastating human diseases. Luminal expansion during organogenesis begins to be elucidated in several systems but we still lack a mechanistic view of the process in many organs. The Drosophila tracheal respiratory system provides an amenable model to study tube size regulation. In the trachea, COPII anterograde transport of luminal proteins is required for extracellular matrix assembly and the concurrent tube expansion. PRINCIPAL FINDINGS: We identified and analyzed Drosophila COPI retrograde transport mutants with narrow tracheal tubes. gammaCOP mutants fail to efficiently secrete luminal components and assemble the luminal chitinous matrix during tracheal tube expansion. Likewise, tube extension is defective in salivary glands, where it also coincides with a failure in the luminal deposition and assembly of a distinct, transient intraluminal matrix. Drosophila gammaCOP colocalizes with cis-Golgi markers and in gammaCOP mutant embryos the ER and Golgi structures are severely disrupted. Analysis of gammaCOP and Sar1 double mutants suggests that bidirectional ER-Golgi traffic maintains the ER and Golgi compartments and is required for secretion and assembly of luminal matrixes during tube expansion. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate the function of COPI components in organ morphogenesis and highlight the common role of apical secretion and assembly of transient organotypic matrices in tube expansion. Intraluminal matrices have been detected in the notochord of ascidians and zebrafish COPI mutants show defects in notochord expansion. Thus, the programmed deposition and growth of distinct luminal molds may provide distending forces during tube expansion in diverse organs. PMID- 18398481 TI - Strain-specific virulence phenotypes of Streptococcus pneumoniae assessed using the Chinchilla laniger model of otitis media. AB - BACKGROUND: Streptococcus pneumoniae [Sp] infection is associated with local and systemic disease. Our current understanding of the differential contributions of genetic strain variation, serotype, and host response to disease phenotype is incomplete. Using the chinchilla model of otitis media [OM] we investigated the disease phenotype generated by the laboratory strain TIGR4 and each of thirteen clinical strains (BS68-75, BS290, BS291, BS293, BS436 and BS437); eleven of the thirteen strains have been genomically sequenced. METHODOLOGY/PRINCIPAL FINDINGS: For each strain 100 colony forming units were injected bilaterally into the tympanic bullae of 6 young adult chinchillas under general anesthesia. All animals were examined daily for local and systemic disease by a blinded observer. Pneumatic otoscopy was used to evaluate local disease, and behavioral assessments served as the measure of systemic disease. Virulence scoring was performed using a 4-point scale to assess four clinical parameters [severity and rapidity of local disease onset; and severity and rapidity of systemic disease onset] during a 10-day evaluation period. Highly significant variation was observed among the strains in their ability to cause disease and moribundity. CONCLUSIONS/SIGNIFICANCE: As expected, there was a significant correlation between the rapidity of systemic disease onset and severity of systemic disease; however, there was little correlation between the severity of otoscopic changes and severity of systemic disease. Importantly, it was observed that different strains of the same serotype produced as broad an array of disease phenotypes as did strains of different serotypes. We attribute these phenotypic differences among the strains to the high degree of genomic plasticity that we have previously documented. PMID- 18398482 TI - The role of the iron transporter ABCB7 in refractory anemia with ring sideroblasts. AB - Refractory Anemia with Ring Sideroblasts (RARS) is an acquired myelodysplastic syndrome (MDS) characterized by an excess iron accumulation in the mitochondria of erythroblasts. The pathogenesis of RARS and the cause of this unusual pattern of iron deposition remain unknown. We considered that the inherited X-linked sideroblastic anemia with ataxia (XLSA/A) might be informative for the acquired disorder, RARS. XLSA/A is caused by partial inactivating mutations of the ABCB7 ATP-binding cassette transporter gene, which functions to enable transport of iron from the mitochondria to the cytoplasm. Furthermore, ABCB7 gene silencing in HeLa cells causes an accumulation of iron in the mitochondria. We have studied the role of ABCB7 in RARS by DNA sequencing, methylation studies, and gene expression studies in primary CD34(+) cells and in cultured erythroblasts. The DNA sequence of the ABCB7 gene is normal in patients with RARS. We have investigated ABCB7 gene expression levels in the CD34(+) cells of 122 MDS cases, comprising 35 patients with refractory anemia (RA), 33 patients with RARS and 54 patients with RA with excess blasts (RAEB), and in the CD34(+) cells of 16 healthy controls. We found that the expression levels of ABCB7 are significantly lower in the RARS group. RARS is thus characterized by lower levels of ABCB7 gene expression in comparison to other MDS subtypes. Moreover, we find a strong relationship between increasing percentage of bone marrow ring sideroblasts and decreasing ABCB7 gene expression levels. Erythroblast cell cultures confirm the low levels of ABCB7 gene expression levels in RARS. These data provide an important link between inherited and acquired forms of sideroblastic anemia and indicate that ABCB7 is a strong candidate gene for RARS. PMID- 18398483 TI - Genomic diversity among Beijing and non-Beijing Mycobacterium tuberculosis isolates from Myanmar. AB - BACKGROUND: The Beijing family of Mycobacterium tuberculosis is dominant in countries in East Asia. Genomic polymorphisms are a source of diversity within the M. tuberculosis genome and may account for the variation of virulence among M. tuberculosis isolates. Till date there are no studies that have examined the genomic composition of M. tuberculosis isolates from the high TB-burden country, Myanmar. METHODOLOGY/PRINCIPLE FINDINGS: Twenty-two M. tuberculosis isolates from Myanmar were screened on whole-genome arrays containing genes from M. tuberculosis H37Rv, M. tuberculosis CDC1551 and M. bovis AF22197. Screening identified 198 deletions or extra regions in the clinical isolates compared to H37Rv. Twenty-two regions differentiated between Beijing and non-Beijing isolates and were verified by PCR on an additional 40 isolates. Six regions (Rv0071-0074 [RD105], Rv1572-1576c [RD149], Rv1585c-1587c [RD149], MT1798-Rv1755c [RD152], Rv1761c [RD152] and Rv0279c) were deleted in Beijing isolates, of which 4 (Rv1572 1576c, Rv1585c-1587c, MT1798-Rv1755c and Rv1761c) were variably deleted among ST42 isolates, indicating a closer relationship between the Beijing and ST42 lineages. The TbD1 region, Mb1582-Mb1583 was deleted in Beijing and ST42 isolates. One M. bovis gene of unknown function, Mb3184c was present in all isolates, except 11 of 13 ST42 isolates. The CDC1551 gene, MT1360 coding for a putative adenylate cyclase, was present in all Beijing and ST42 isolates (except 1). The pks15/1 gene, coding for a putative virulence factor, was intact in all Beijing and non-Beijing isolates, except in ST42 and ST53 isolates. CONCLUSION: This study describes previously unreported deletions/extra regions in Beijing and non-Beijing M. tuberculosis isolates. The modern and highly frequent ST42 lineage showed a closer relationship to the hypervirulent Beijing lineage than to the ancient non-Beijing lineages. The pks15/1 gene was disrupted only in modern non Beijing isolates. This is the first report of an in-depth analysis on the genomic diversity of M. tuberculosis isolates from Myanmar. PMID- 18398484 TI - Altered dopamine signaling in naturally occurring maternal neglect. AB - BACKGROUND: Child neglect is the most common form of child maltreatment, yet the biological basis of maternal neglect is poorly understood and a rodent model is lacking. METHODOLOGY/PRINCIPAL FINDINGS: The current study characterizes a population of mice (MaD1) which naturally exhibit maternal neglect (little or no care of offspring) at an average rate of 17% per generation. We identified a set of risk factors that can predict future neglect of offspring, including decreased self-grooming and elevated activity. At the time of neglect, neglectful mothers swam significantly more in a forced swim test relative to nurturing mothers. Cross-fostered offspring raised by neglectful mothers in turn exhibit increased expression of risk factors for maternal neglect and decreased maternal care as adults, suggestive of possible epigenetic contributions to neglect. Unexpectedly, offspring from neglectful mothers elicited maternal neglect from cross-fostered nurturing mothers, suggesting that factors regulating neglect are not solely within the mother. To identify a neurological pathway underlying maternal neglect, we examined brain activity in neglectful and nurturing mice. c-Fos expression was significantly elevated in neglectful relative to nurturing mothers in the CNS, particularly within dopamine associated areas, such as the zona incerta (ZI), ventral tegmental area (VTA), and nucleus accumbens. Phosphorylated tyrosine hydroxylase (a marker for dopamine production) was significantly elevated in ZI and higher in VTA (although not significantly) in neglectful mice. Tyrosine hydroxylase levels were unaltered, suggesting a dysregulation of dopamine activity rather than cell number. Phosphorylation of DARPP-32, a marker for dopamine D1-like receptor activation, was elevated within nucleus accumbens and caudate-putamen in neglectful versus nurturing dams. CONCLUSIONS/SIGNIFICANCE: These findings suggest that atypical dopamine activity within the maternal brain, especially within regions involved in reward, is involved in naturally occurring neglect and that MaD1 mice are a useful model for understanding the basis of naturally occurring neglect. PMID- 18398485 TI - The CD85j+ NK cell subset potently controls HIV-1 replication in autologous dendritic cells. AB - Natural killer (NK) cells and dendritic cells (DC) are thought to play critical roles in the first phases of HIV infection. In this study, we examined changes in the NK cell repertoire and functions occurring in response to early interaction with HIV-infected DC, using an autologous in vitro NK/DC coculture system. We show that NK cell interaction with HIV-1-infected autologous monocyte-derived DC (MDDC) modulates NK receptor expression. In particular, expression of the CD85j receptor on NK cells was strongly down-regulated upon coculture with HIV-1 infected MDDC. We demonstrate that CD85j(+) NK cells exert potent control of HIV 1 replication in single-round and productively HIV-1-infected MDDC, whereas CD85j(-) NK cells induce a modest and transient decrease of HIV-1 replication. HIV-1 suppression in MDCC by CD85j(+) NK cells required cell-to-cell contact and did not appear mediated by cytotoxicity or by soluble factors. HIV-1 inhibition was abolished when NK-MDDC interaction through the CD85j receptor was blocked with a recombinant CD85j molecule, whereas inhibition was only slightly counteracted by blocking HLA class I molecules, which are known CD85j ligands. After masking HLA class I molecules with specific antibodies, a fraction of HIV-1 infected MDDC was still strongly stained by a recombinant CD85j protein. These results suggest that CD85j(+) NK cell inhibition of HIV-1 replication in MDDC is mainly mediated by CD85j interaction with an unknown ligand (distinct from HLA class I molecules) preferentially expressed on HIV-1-infected MDDC. PMID- 18398486 TI - Inactivation of [Fe-S] metalloproteins mediates nitric oxide-dependent killing of Burkholderia mallei. AB - BACKGROUND: Much remains to be known about the mechanisms by which O(2)-dependent host defenses mediate broad antimicrobial activity. METHODOLOGY/PRINCIPAL FINDINGS: We show herein that reactive nitrogen species (RNS) generated by inducible nitric oxide (NO) synthase (iNOS) account for the anti-Burkholderia mallei activity of IFNgamma-primed macrophages. Inducible NOS-mediated intracellular killing may represent direct bactericidal activity, because B. mallei showed an exquisite sensitivity to NO generated chemically. Exposure of B. mallei to sublethal concentrations of NO upregulated transcription of [Fe-S] cluster repair genes, while damaging the enzymatic activity of the [Fe-S] protein aconitase. To test whether [Fe-S] clusters are critical targets for RNS-dependent killing of B. mallei, a mutation was constructed in the NO-induced, [Fe-S] cluster repair regulator iscR. Not only was the iscR mutant hypersusceptible to iNOS-mediated killing, but its aconitase pool was readily oxidized by NO donors as compared to wild-type controls. Although killed by authentic H(2)O(2), which also oxidizes [Fe-S] clusters, B. mallei appear to be resilient to NADPH oxidase mediated cytotoxicity. The poor respiratory burst elicited by this bacterium likely explains why the NADPH oxidase is nonessential to the killing of B. mallei while it is still confined within phagosomes. CONCLUSIONS/SIGNIFICANCE: Collectively, these findings have revealed a disparate role for NADPH oxidase and iNOS in the innate macrophage response against the strict aerobe B. mallei. To the best of our knowledge, this is the first instance in which disruption of [Fe S] clusters is demonstrated as cause of the bactericidal activity of NO congeners. PMID- 18398487 TI - Use of short tandem repeat sequences to study Mycobacterium leprae in leprosy patients in Malawi and India. AB - BACKGROUND: Inadequate understanding of the transmission of Mycobacterium leprae makes it difficult to predict the impact of leprosy control interventions. Genotypic tests that allow tracking of individual bacterial strains would strengthen epidemiological studies and contribute to our understanding of the disease. METHODOLOGY/PRINCIPAL FINDINGS: Genotyping assays based on variation in the copy number of short tandem repeat sequences were applied to biopsies collected in population-based epidemiological studies of leprosy in northern Malawi, and from members of multi-case households in Hyderabad, India. In the Malawi series, considerable genotypic variability was observed between patients, and also within patients, when isolates were collected at different times or from different tissues. Less within-patient variability was observed when isolates were collected from similar tissues at the same time. Less genotypic variability was noted amongst the closely related Indian patients than in the Malawi series. CONCLUSIONS/SIGNIFICANCE: Lineages of M. leprae undergo changes in their pattern of short tandem repeat sequences over time. Genetic divergence is particularly likely between bacilli inhabiting different (e.g., skin and nerve) tissues. Such variability makes short tandem repeat sequences unsuitable as a general tool for population-based strain typing of M. leprae, or for distinguishing relapse from reinfection. Careful use of these markers may provide insights into the development of disease within individuals and for tracking of short transmission chains. PMID- 18398488 TI - Differential gene expression changes in children with severe dengue virus infections. AB - BACKGROUND: The host response to dengue virus infection is characterized by the production of numerous cytokines, but the overall picture appears to be complex. It has been suggested that a balance may be involved between protective and pathologic immune responses. This study aimed to define differential immune responses in association with clinical outcomes by gene expression profiling of a selected panel of inflammatory genes in whole blood samples from children with severe dengue infections. METHODOLOGY/PRINCIPAL FINDINGS: Whole blood mRNA from 56 Indonesian children with severe dengue virus infections was analyzed during early admission and at day -1, 0, 1, and 5-8 after defervescence. Levels were related to baseline levels collected at a 1-month follow-up visit. Processing of mRNA was performed in a single reaction by multiplex ligation-dependent probe amplification, measuring mRNA levels from genes encoding 36 inflammatory proteins and 14 Toll-like receptor (TLR)-associated molecules. The inflammatory gene profiles showed up-regulation during infection of eight genes, including IFNG and IL12A, which indicated an antiviral response. On the contrary, genes associated with the nuclear factor (NF)-kappaB pathway were down-regulated, including NFKB1, NFKB2, TNFR1, IL1B, IL8, and TNFA. Many of these NF-kappaB pathway-related genes, but not IFNG or IL12A, correlated with adverse clinical events such as development of pleural effusion and hemorrhagic manifestations. The TLR profile showed that TLRs were differentially activated during severe dengue infections: increased expression of TLR7 and TLR4R3 was found together with a decreased expression of TLR1, TLR2, TLR4R4, and TLR4 co-factor CD14. CONCLUSIONS/SIGNIFICANCE: These data show that different immunological pathways are differently expressed and associated with different clinical outcomes in children with severe dengue infections. PMID- 18398489 TI - Differential release and phagocytosis of tegument glycoconjugates in neurocysticercosis: implications for immune evasion strategies. AB - Neurocysticercosis (NCC) is an infection of the central nervous system (CNS) by the metacestode of the helminth Taenia solium. The severity of the symptoms is associated with the intensity of the immune response. First, there is a long asymptomatic period where host immunity seems incapable of resolving the infection, followed by a chronic hypersensitivity reaction. Since little is known about the initial response to this infection, a murine model using the cestode Mesocestoides corti (syn. Mesocestoides vogae) was employed to analyze morphological changes in the parasite early in the infection. It was found that M. corti material is released from the tegument making close contact with the nervous tissue. These results were confirmed by infecting murine CNS with ex vivo labeled parasites. Because more than 95% of NCC patients exhibit humoral responses against carbohydrate-based antigens, and the tegument is known to be rich in glycoconjugates (GCs), the expression of these types of molecules was analyzed in human, porcine, and murine NCC specimens. To determine the GCs present in the tegument, fluorochrome-labeled hydrazides as well as fluorochrome labeled lectins with specificity to different carbohydrates were used. All the lectins utilized labeled the tegument. GCs bound by isolectinB4 were shed in the first days of infection and not resynthesized by the parasite, whereas GCs bound by wheat germ agglutinin and concavalinA were continuously released throughout the infectious process. GCs bound by these three lectins were taken up by host cells. Peanut lectin-binding GCs, in contrast, remained on the parasite and were not detected in host cells. The parasitic origin of the lectin-binding GCs found in host cells was confirmed using antibodies against T. solium and M. corti. We propose that both the rapid and persistent release of tegumental GCs plays a key role in the well-known immunomodulatory effects of helminths, including immune evasion and life-long inflammatory sequelae seen in many NCC patients. PMID- 18398490 TI - Tissue invasion by Entamoeba histolytica: evidence of genetic selection and/or DNA reorganization events in organ tropism. AB - Entamoeba histolytica infection may have various clinical manifestations. Nine out of ten E. histolytica infections remain asymptomatic, while the remainder become invasive and cause disease. The most common form of invasive infection is amebic diarrhea and colitis, whereas the most common extra-intestinal disease is amebic liver abscess. The underlying reasons for the different outcomes are unclear, but a recent study has shown that the parasite genotype is a contributor. To investigate this link further we have examined the genotypes of E. histolytica in stool- and liver abscess-derived samples from the same patients. Analysis of all 18 paired samples (16 from Bangladesh, one from the United States of America, and one from Italy) revealed that the intestinal and liver abscess amebae are genetically distinct. The results suggest either that E. histolytica subpopulations in the same infection show varying organ tropism, or that a DNA reorganization event takes place prior to or during metastasis from intestine to liver. PMID- 18398491 TI - Susceptibility to Vibrio cholerae infection in a cohort of household contacts of patients with cholera in Bangladesh. AB - BACKGROUND: Despite recent progress in understanding the molecular basis of Vibrio cholerae pathogenesis, there is relatively little knowledge of the factors that determine the variability in human susceptibility to V. cholerae infection. METHODS AND FINDINGS: We performed an observational study of a cohort of household contacts of cholera patients in Bangladesh, and compared the baseline characteristics of household members who went on to develop culture-positive V. cholerae infection with individuals who did not develop infection. Although the vibriocidal antibody is the only previously described immunologic marker associated with protection from V. cholerae infection, we found that levels of serum IgA specific to three V. cholerae antigens-the B subunit of cholera toxin, lipopolysaccharide, and TcpA, the major component of the toxin-co-regulated pilus also predicted protection in household contacts of patients infected with V. cholerae O1, the current predominant cause of cholera. Circulating IgA antibodies to TcpA were also associated with protection from V. cholerae O139 infection. In contrast, there was no association between serum IgG antibodies specific to these three antigens and protection from infection with either serogroup. We also found evidence that host genetic characteristics and serum retinol levels modify susceptibility to V. cholerae infection. CONCLUSIONS: Our observation that levels of serum IgA (but not serum IgG) directed at certain V. cholerae antigens are associated with protection from infection underscores the need to better understand anti-V. cholerae immunity at the mucosal surface. Furthermore, our data suggest that susceptibility to V. cholerae infection is determined by a combination of immunologic, nutritional, and genetic characteristics; additional factors that influence susceptibility to cholera remain unidentified. PMID- 18398492 TI - Indian medicinal mushrooms as a source of antioxidant and antitumor agents. AB - Medicinal mushrooms occurring in South India namely Ganoderma lucidum, Phellinus rimosus, Pleurotus florida and Pleurotus pulmonaris possessed profound antioxidant and antitumor activities. This indicated that these mushrooms would be valuable sources of antioxidant and antitumor compounds. Investigations also revealed that they had significant antimutagenic and anticarcinogenic activities. Thus, Indian medicinal mushrooms are potential sources of antioxidant and anticancer compounds. However, intensive and extensive investigations are needed to exploit their valuable therapeutic use. PMID- 18398494 TI - Physico-chemical studies on the evaluation of the antioxidant activity of herbal extracts and active principles of some Indian medicinal plants. AB - Understanding of the efficacy and mechanism for the reaction of the biologically important radicals with natural and/or synthetic antioxidants is the first step towards the development of future therapeutic agents. The kinetic parameters e.g., formation and decay rate constants predict the efficacy of an antioxidant and its fate after reaction. These parameters also dictate the ease with which competing reactions would occur in a bio-environment. The spectroscopic parameters provide the clue to the site of free radical attack to these antioxidants. Here, in this article an attempt has been made to show the use of physico-chemical methods in the evaluation of antioxidant activity of some important medicinal plants commonly used in India and the subcontinent. The systems chosen here for discussions are herbal extracts as such, curcumin from turmeric, methoxy phenols from Indian spices, dehydrogingerdione from ginger and bakuchiol from Psoralea corylifolia. All the examples shown in this article illustrate the potential of the pulse radiolysis coupled with kinetic spectroscopy and other physicochemical techniques for the study of antioxidants either in the form of mixture as in herbal extract or as an isolated compound. PMID- 18398493 TI - Indian herbs and herbal drugs used for the treatment of diabetes. AB - Traditional Medicines derived from medicinal plants are used by about 60% of the world's population. This review focuses on Indian Herbal drugs and plants used in the treatment of diabetes, especially in India. Diabetes is an important human ailment afflicting many from various walks of life in different countries. In India it is proving to be a major health problem, especially in the urban areas. Though there are various approaches to reduce the ill effects of diabetes and its secondary complications, herbal formulations are preferred due to lesser side effects and low cost. A list of medicinal plants with proven antidiabetic and related beneficial effects and of herbal drugs used in treatment of diabetes is compiled. These include, Allium sativum, Eugenia jambolana, Momordica charantia Ocimum sanctum, Phyllanthus amarus, Pterocarpus marsupium, Tinospora cordifolia, Trigonella foenum graecum and Withania somnifera. One of the etiologic factors implicated in the development of diabetes and its complications is the damage induced by free radicals and hence an antidiabetic compound with antioxidant properties would be more beneficial. Therefore information on antioxidant effects of these medicinal plants is also included. PMID- 18398495 TI - Mechanism of Liver Injury during Obstructive Jaundice: Role of Nitric Oxide, Splenic Cytokines, and Intestinal Flora. AB - To elucidate the roles of enteric bacteria and immunological interactions among liver, spleen and intestine in the pathogenesis of liver injury during obstructive jaundice, we studied the effects of antibiotics and splenectomy on bile-duct-ligated C57BL mice. When animals were subjected to bile-duct-ligation (BDL), plasma levels of bilirubin, alanine aminotransferase and aspartate aminotransferase increased markedly. However, the increases in plasma transaminases were significantly lower in splenectomized or antibiotics-treated groups than in the control BDL group. Histological examination revealed that liver injury was also low in the two groups. BDL markedly increased plasma level of interferon-gamma (IFN-gamma) and the expression of inducible nitric oxide synthase (iNOS) in liver and spleen. These changes were suppressed either by splenectomy or administration of antibiotics. Kinetic analysis revealed that BDL induced liver injury and the increase of interleukin-10 (IL-10) and INF-gamma were lower in iNOS(-/-) than in wild type animals. BDL markedly increased the expression of IgA in colonic mucosa. These observations suggest that enteric bacteria, nitric oxide and cytokines including IFN-gamma and IL-10 derived from spleen and intestines form a critical network that determines the extent of liver injury during obstructive jaundice. PMID- 18398496 TI - Protective effects of coenzyme q(10) on decreased oxidative stress resistance induced by simvastatin. AB - The effects of simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl CoA reductase (HMG-CoA reductase), on oxidative stress resistance and the protective effects of coenzyme Q (CoQ) were investigated. When simvastatin was administered orally to mice, the levels of oxidized and reduced CoQ(9) and CoQ(10) in serum, liver, and heart, decreased significantly when compared to those of control. The levels of thiobarbituric acid reactive substances induced by Fe(2+)-ascorbate in liver and heart mitochondria also increased significantly with simvastatin. Furthermore, cultured cardiac myocytes treated with simvastatin exhibited less resistance to oxidative stress, decreased time to the cessation of spontaneous beating in response to H(2)O(2) addition, and decreased responsiveness to electrical field stimulation. These results suggested that oral administration of simvastatin suppresses the biosynthesis of CoQ, which shares the same biosynthesis pathway as cholesterol up to farnesyl pyrophosphate, thus compromising the physiological function of reduced CoQ, which possesses antioxidant activity. However, these undesirable effects induced by simvastatin were alleviated by coadministering CoQ(10) with simvastatin to mice. Simvastatin also reduced the activity of NADPH-CoQ reductase, a biological enzyme that converts oxidized CoQ to the corresponding reduced CoQ, while CoQ(10) administration improved it. These findings may also support the efficacy of coadministering CoQ(10) with statins. PMID- 18398497 TI - Genetic and epigenetic markers to identify high risk patients for multiple early gastric cancers after treatment with endoscopic mucosal resection. AB - The development of multiple gastric cancer is a major problem after the endoscopic resection of the first early gastric cancer. To find out markers to identify high risk patients, we analyzed the microsatellite instability (MSI) status and hypermethylation of tumor-related genes in multiple gastric cancers. Sixty-four adenocarcinomas resected by endoscopy, including 32 early solitary gastric cancers (SGCs) from 32 patients and 32 multiple gastric cancers (MGCs) from 14 patients, were employed. We analyzed MSI and the methylation status of promoter regions of the hMLH1, MGMT, p16 and E-cadherin using methylation specific Polymerase Chain Reaction. Expression levels of hMLH1 were examined by immunohistochemistry. MSI (+) was detected in 5 of the 14 (35.7%) patients with MGCs, and in only 3 of the 32 patients (9.3%) with SGCs. Significant differences were observed between the 2 groups (p<0.001). Hypermethylation of hMLH1 was more frequently detected in MGCs than in SGCs (p<0.01), whereas significant difference was not observed in the frequency of MGMT, p16 or E-cadherin promoter methylation between the 2 groups. In conclusion, our results indicate that inactivation of hMLH1 through promoter hypermethylation may be involved in the development of multiple gastric cancers following the MSI pathway. PMID- 18398499 TI - Changes in lymphocyte phenotypes and cytokine production by surgical stress in a rat small intestinal resection model. AB - Small intestinal resection rats are used widely as a malabsorption model, but the immunological changes are unclear. We examined the changes in systemic and mucosal immune status after a small intestinal resection in rats with a controlled nutritional status. Rats had 60% of their small intestine removed. At 5 days after the surgery, spleen cells and intraepithelial lymphocytes (IEL) were isolated. The phenotypes of spleen cells and IEL, the production patterns of Th1 and Th2 cytokines, and the proinflammatory cytokine levels in the plasma were measured. CD4+ T cells in the blood and spleen were significantly decreased in the Resection group (p<0.05). In contrast, IEL subpopulations were not different between the two groups. Interferon-gamma production from the spleen cells was significantly decreased in the Resection group (p<0.05). Interleukin (IL)-4 production was not different between the two groups. Plasma IL-6 concentrations were significantly elevated in the Resection group 6 h after surgery (p<0.05). In conclusions, small intestinal resection in rats suppressed systemic immunity, and this model is useful as a surgical stress model. PMID- 18398498 TI - Analysis of Beta-Tubulin Gene Exon 4 Mutations in Advanced Stage III or IV Gastric Cancer. AB - The mechanisms that cause chemoresistance of gastric cancer have yet to be elucidated. Taxanes and promising agents that were recently approved for treatment of advanced or recurrent gastric cancer. Mutations of beta-tubulin, which is a target of taxianes, have been shown to confer chemoresistance against these agents. The aim of the present study is to investigate the presence of mutations of the beta-tubulin in gastric cancer tissues. Sixty-six patients with advanced stage III or IV gastric cancer patients enrolled in this study. Paired samples of gastric cancer tissue and normal mucosa were obtained by endoscopy. The guanosine 5'-triphosphate (GTP)-binding site in exon 4 of the beta-tubulin gene was examined by polymerase chain reaction single-strand conformational polymorphism (PCR-SSCP) analysis, followed by sequencing of the products with abnormally shifted bands. SSCP analysis showed abnormal bands upstream of the GTP binding site in 7 of the 66 patients, but sequence analysis found no nucleotide substitutions in these patients. Three variant bands were also detected down stream of the the GTP-binding site, but the sequences of the 3 products corresponded to those of two independent pseudogenes. Thus, none of the tumor samples showed mutation of the beta-tubulin exon 4 GTP-binding site. In conclusion, these findings suggest that mutations of the beta-tubulin gene are rare and are unlikely to be an important cause of taxane resistance to taxians. PMID- 18398500 TI - Effects of prolonged high phosphorus diet on phosphorus and calcium balance in rats. AB - The amount of phosphorus contained in food as food additives is currently increasing and a high intake of phosphorus can cause various diseases. To determine the effects of a prolonged high phosphorus diet, here we investigated the phosphorus and calcium balance and expression of type IIa sodium-dependent phosphate transporter (Npt IIa) in mature rats. Wistar male rats (8-weeks old) were divided into five groups and fed diets containing 0.6% calcium plus 0.3, 0.6, 0.9, 1.2 or 1.5% phosphorus for 4 weeks. Urinary and fecal phosphorus excretions were significantly increased by the high phosphorus diets (from 0.6 to 1.5%), dependent on the amount of dietary phosphorus. The net absorption of intestinal phosphorus was also significantly increased by high phosphorus diets. As a result, a negative phosphorus balance was observed in rats given the 1.2% or 1.5% phosphorus diets. Serum parathyroid hormone and 1,25-dihydroxyvitamin D(3) concentrations were increased by high phosphorus diets. In addition, high phosphorus diets decreased the expression of Npt IIa mRNA and protein in the renal brush border membrane. Taken together, these results suggest that diets containing 1.2 or 1.5% phosphorus plus 0.6% calcium have potentially adverse effects on phosphorus homeostasis in mature rat. PMID- 18398501 TI - Effects of Oral Administration of Moringa oleifera Lam on Glucose Tolerance in Goto-Kakizaki and Wistar Rats. AB - Medicinal plants constitute an important source of potential therapeutic agents for diabetes. In the present study, we investigated the effects of Moringa oleifera (MO) Lam, Moringacea, on glucose tolerance in Wistar rats and Goto Kakizaki (GK) rats, modeled type 2 diabetes. Major polyphenols in MO powder were quercetin glucosides, rutin, kaempferol glycosides and chlorogenic acids by HPLC analysis. As the results of glucose tolerance test, MO significantly decreased the blood glucose at 20, 30, 45and 60 min for GK rats and at 10, 30 and 45 min for Wistar rats (p<0.05) compared to the both controls after glucose administration. The area under the curve of changes in the blood glucose was significantly higher in the GK control group than in the GK plus MO group (p<0.05) in the periods 30-60 min and 60-120 min. Furthermore, MO significantly decreased stomach emptying in GK rats (p<0.05). The results indicated that MO has an ameliorating effect for glucose intolerance, and the effect might be mediated by quercetin-3-glucoside and fiber contents in MO leaf powder. The action of MO was greater in GK rats than in Wistar rats. PMID- 18398502 TI - The Anti-Obesity Effect of the Palatinose-Based Formula Inslow is Likely due to an Increase in the Hepatic PPAR-alpha and Adipocyte PPAR-gamma Gene Expressions. AB - Abdominal obesity is a principal risk factor in the development of metabolic syndrome. Previously, we showed that a palatinose-based liquid formula, Inslow/MHN-01, suppressed postprandial plasma glucose level and reduced visceral fat accumulation better than the standard formula (SF). To elucidate the mechanism of Inslow-mediated anti-obesity effect, expression levels of genes involved in the glucose and lipid metabolism were compared in Inslow- and SF-fed rats. Both fasting plasma insulin level and average islet sizes were reduced in the Inslow group. We also found less abdominal fat accumulation and reduced hepatic triacylglycerol content in the Inslow group. Expression of the beta oxidation enzymes and uncoupling potein-2 (UCP-2) mRNAs in the liver of the Inslow group were higher than the SF group, which was due to a concomitant higher expression of the peroxisome proliferator-activated receptor (PPAR)-alpha mRNA in the former. Furthermore, expression of the UCP-2 and adiponectin mRNAs in the epididymal fat were higher in the Inslow group than the SF group, and were stimulated by a concomitant increase of the PPAR-gamma gene expression in the former. These results strongly suggested that the anti-obesity effect of Inslow was due to an increase in the hepatic PPAR-alpha and adipocyte PPAR-gamma gene expressions. PMID- 18398503 TI - BRAF gene duplication constitutes a mechanism of MAPK pathway activation in low grade astrocytomas. AB - The molecular pathogenesis of pediatric astrocytomas is still poorly understood. To further understand the genetic abnormalities associated with these tumors, we performed a genome-wide analysis of DNA copy number aberrations in pediatric low grade astrocytomas by using array-based comparative genomic hybridization. Duplication of the BRAF protooncogene was the most frequent genomic aberration, and tumors with BRAF duplication showed significantly increased mRNA levels of BRAF and a downstream target, CCND1, as compared with tumors without duplication. Furthermore, denaturing HPLC showed that activating BRAF mutations were detected in some of the tumors without BRAF duplication. Similarly, a marked proportion of low-grade astrocytomas from adult patients also had BRAF duplication. Both the stable silencing of BRAF through shRNA lentiviral transduction and pharmacological inhibition of MEK1/2, the immediate downstream phosphorylation target of BRAF, blocked the proliferation and arrested the growth of cultured tumor cells derived from low-grade gliomas. Our findings implicate aberrant activation of the MAPK pathway due to gene duplication or mutation of BRAF as a molecular mechanism of pathogenesis in low-grade astrocytomas and suggest inhibition of the MAPK pathway as a potential treatment. PMID- 18398504 TI - Vascular targeting of anti-CD40 antibodies and IL-2 into autochthonous tumors enhances immunotherapy in mice. AB - Current anticancer therapy is a delicate balance between elimination of malignant cells and harmful side effects for the host. In this study, we used a tumor homing peptide to engineer anti-CD40 agonist antibodies and recombinant IL-2 such that they were selectively delivered into spontaneously arising tumors in a transgenic mouse model of islet cell carcinogenesis. Intravenous injection of these agents, either separately or together, led to accumulation in the vicinity of tumor neovessels without toxic side effects. Although both molecules are critical for adaptive immunity, the most profound effects were seen in endothelial cells. Combined, local anti-CD40 and IL-2 therapy reduced tumor vascularity and significantly delayed tumor growth in mice. Remarkably, tumor bearing mice remained disease-free long-term when targeted anti-CD40 and IL-2 were combined with transfers of preactivated antitumor immune cells. In this therapeutic setting, triggering of CD40 on endothelial cells induced an inflammatory response of the vessel wall and facilitated effector cell accumulation in the tumor parenchyma while IL-2 promoted antigen-specific immune cell persistence. We believe this is a novel and highly effective anticancer approach, whereby tumor stroma is "conditioned" for enhanced immune cell entry and survival, facilitating immune-mediated tumor destruction and leading to a sustained antitumor response. PMID- 18398505 TI - Long-term expression of murine activated factor VII is safe, but elevated levels cause premature mortality. AB - Intravenous infusion of recombinant human activated Factor VII (FVIIa) has been used for over a decade in the successful management of bleeding episodes in patients with inhibitory antibodies to Factor VIII or Factor IX. Previously, we showed that expression of murine FVIIa (mFVIIa) from an adeno-associated viral (AAV) vector corrected abnormal hemostatic parameters in hemophilia B mice. To pursue this as a therapeutic approach, we sought to define safe and effective levels of FVIIa for continuous expression. In mice transgenic for mFVIIa or injected with AAV-mFVIIa, we analyzed survival, expression levels, in vitro and in vivo coagulation tests, and histopathology for up to 16 months after birth/mFVIIa expression. We found that continuous expression of mFVIIa at levels at or below 1.5 microg/ml was safe, effective, and compatible with a normal lifespan. However, expression levels of 2 microg/ml or higher were associated with thrombosis and early mortality, with pathologic findings in the heart and lungs that were rescued in a low-factor X (low-FX) mouse background, suggesting a FX-mediated effect. The findings from these mouse models of continuous FVIIa expression have implications for the development of a safe gene transfer approach for hemophilia and are consistent with the possibility of thromboembolic risk of continuously elevated FVIIa levels. PMID- 18398506 TI - TRPA1 is a major oxidant sensor in murine airway sensory neurons. AB - Sensory neurons in the airways are finely tuned to respond to reactive chemicals threatening airway function and integrity. Nasal trigeminal nerve endings are particularly sensitive to oxidants formed in polluted air and during oxidative stress as well as to chlorine, which is frequently released in industrial and domestic accidents. Oxidant activation of airway neurons induces respiratory depression, nasal obstruction, sneezing, cough, and pain. While normally protective, chemosensory airway reflexes can provoke severe complications in patients affected by inflammatory airway conditions like rhinitis and asthma. Here, we showed that both hypochlorite, the oxidizing mediator of chlorine, and hydrogen peroxide, a reactive oxygen species, activated Ca(2+) influx and membrane currents in an oxidant-sensitive subpopulation of chemosensory neurons. These responses were absent in neurons from mice lacking TRPA1, an ion channel of the transient receptor potential (TRP) gene family. TRPA1 channels were strongly activated by hypochlorite and hydrogen peroxide in primary sensory neurons and heterologous cells. In tests of respiratory function, Trpa1(-/-) mice displayed profound deficiencies in hypochlorite- and hydrogen peroxide-induced respiratory depression as well as decreased oxidant-induced pain behavior. Our results indicate that TRPA1 is an oxidant sensor in sensory neurons, initiating neuronal excitation and subsequent physiological responses in vitro and in vivo. PMID- 18398507 TI - Antibody association with HER-2/neu-targeted vaccine enhances CD8 T cell responses in mice through Fc-mediated activation of DCs. AB - The pathogenic nature of cancer is attributed, at least in part, to the ability of tumors cells to induce systemic and local mechanisms of immune tolerance. However, we previously reported that tumor-free survival in up to 100% of tolerized HER-2/neu transgenic mice can be achieved by administration of neu specific mAb concurrently with a HER-2/neu-expressing, GM-CSF-secreting whole cell vaccine. In this report, we show that one mechanism of improved antitumor activity induced by the combination of these 2 neu-targeted interventions was enhanced Fc-mediated activation of APCs. Specifically, in vivo studies demonstrated localization of radiolabeled neu-specific mAb at the vaccine site. Subsequently, increased accumulation of neu-specific mAb at the vaccine-draining lymph node correlated with increased vaccine cell uptake by DCs in vivo. This led to enhancement of CD8(+) neu-specific T cell function in terms of proliferation, cytokine production, and central memory development. Thus, the administration of a neu-specific mAb with a neu-targeted GM-CSF-secreting tumor vaccine enhanced induction of neu-specific CD8(+) T cells through Fc-mediated activation of DCs. This multimodality attack on the same tumor antigen may have the potential to overcome tolerance to self antigens and weaken the immunosuppressive networks within the tumor microenvironment. PMID- 18398508 TI - Bim-mediated deletion of antigen-specific CD8 T cells in patients unable to control HBV infection. AB - HBV-specific CD8(+) T cells are critical for a successful immune response to HBV infection. They are markedly diminished in number in patients who fail to control the virus, but the mechanisms resulting in their depletion remain ill defined. Here, we dissected the defective HBV-specific CD8(+) T cell response associated with chronic HBV infection by gene expression profiling. We found that HBV specific CD8(+) T cells from patients with different clinical outcomes could be distinguished by their patterns of gene expression. Microarray analysis revealed that overlapping clusters of functionally related apoptotic genes were upregulated in HBV-specific CD8(+) T cells from patients with chronic compared with resolved infection. Further analysis confirmed that levels of the proapoptotic protein Bcl2-interacting mediator (Bim) were upregulated in HBV specific CD8(+) T cells from patients with chronic HBV infection. Blocking Bim mediated apoptosis enhanced recovery of HBV-specific CD8(+) T cells both in culture and directly ex vivo. Consistent with evidence that Bim mediates apoptosis of CD8(+) T cells expressing low levels of CD127 (IL-7R), the few surviving HBV-specific CD8(+) T cells were CD127(hi )and had elevated levels of the antiapoptotic protein Mcl1, suggesting they were amenable to IL-7-mediated rescue from apoptosis. We therefore postulate that Bim-mediated attrition of HBV specific CD8(+) T cells contributes to the inability of these cell populations to persist and control viral replication. PMID- 18398509 TI - Congenital myasthenia-related AChR delta subunit mutation interferes with intersubunit communication essential for channel gating. AB - Congenital myasthenias (CMs) arise from defects in neuromuscular junction associated proteins. Deciphering the molecular bases of the CMs is required for therapy and illuminates structure-function relationships in these proteins. Here, we analyze the effects of a mutation in 1 of 4 homologous subunits in the AChR from a CM patient, a Leu to Pro mutation at position 42 of the delta subunit. The mutation is located in a region of contact between subunits required for rapid opening of the AChR channel and impedes the rate of channel opening. Substitutions of Gly, Lys, or Asp for deltaL42, or substitutions of Pro along the local protein chain, also slowed channel opening. Substitution of Pro for Leu in the epsilon subunit slowed opening, whereas this substitution had no effect in the beta subunit and actually sped opening in the alpha subunit. Analyses of energetic coupling between residues at the subunit interface showed that deltaL42 is functionally linked to alphaT127, a key residue in the adjacent alpha subunit required for rapid channel opening. Thus, deltaL42 is part of an intersubunit network that enables ACh binding to rapidly open the AChR channel, which may be compromised in patients with CM. PMID- 18398510 TI - Impaired microRNA processing causes corpus luteum insufficiency and infertility in mice. AB - The microRNA (miRNA) processing enzyme Dicer1 is required for zygotic and embryonic development, but the early embryonic lethality of Dicer1 null alleles in mice has limited our ability to address the role of Dicer1 in normal mouse growth and development. To address this question, we used a mouse mutant with a hypomorphic Dicer1 allele (Dicer(d/d)) and found that Dicer1 deficiency resulted in female infertility. This defect in female Dicer(d/d) mice was caused by corpus luteum (CL) insufficiency and resulted, at least in part, from the impaired growth of new capillary vessels in the ovary. We found that the impaired CL angiogenesis in Dicer(d/d) mice was associated with a lack of miR17-5p and let7b, 2 miRNAs that participate in angiogenesis by regulating the expression of the antiangiogenic factor tissue inhibitor of metalloproteinase 1. Furthermore, injection of miR17-5p and let7b into the ovaries of Dicer(d/d) mice partially normalized tissue inhibitor of metalloproteinase 1 expression and CL angiogenesis. Our data indicate that the development and function of the ovarian CL is a physiological process that appears to be regulated by miRNAs and requires Dicer1 function. PMID- 18398512 TI - Not allergic to life. AB - A lot of things have changed over the 3 years since I first wrote for Australian Family Physician; but one thing remains... without prompt medical assistance I will die if I eat a nut. PMID- 18398511 TI - Survival of lethal poxvirus infection in mice depends on TLR9, and therapeutic vaccination provides protection. AB - Poxviruses such as the causative agent of smallpox have developed multiple strategies to suppress immune responses, including the suppression of DC activation. Since poxviruses are large DNA viruses, we hypothesized that their detection by DCs may involve the endosomal DNA recognition receptor TLR9. Indeed, we have shown here that DC recognition of ectromelia virus (ECTV), the causative agent of mousepox, completely depended on TLR9. The importance of TLR9 was highlighted by the fact that mice lacking TLR9 showed drastically increased susceptibility to infection with ECTV. In contrast, we found that the strongly attenuated poxvirus modified vaccinia virus Ankara (MVA) activated DCs by both TLR9-dependent and -independent pathways. We therefore tested whether we could use the broader induction of immune responses by MVA to protect mice from a lethal infection with ECTV. Indeed, MVA given at the same time as a lethal dose of ECTV protected mice from death. Importantly, MVA also rescued TLR9-deficient mice if administered 2 full days after an otherwise lethal infection with ECTV. Therefore, these data suggest an essential role for TLR9 in the defense against poxviruses. In addition, postexposure application of MVA may protect against lethal poxvirus infection. PMID- 18398513 TI - Allergic rhinitis. AB - The BEACH program (Bettering the Evaluation and Care of Health) shows that allergic rhinitis was managed six times per 1000 encounters between October 2002 to September 2007. This suggests that general practitioners manage allergic rhinitis about 619 000 times nationally each year. PMID- 18398514 TI - Allergy prevention - Current recommendations and new insights. AB - BACKGROUND: The prevalence of allergic disease has increased considerably in recent decades and Australia has one of the highest rates of allergic disease in the world. As there is currently no cure for allergic diseases, prevention offers a logical approach to addressing the rising burden of disease. The factors responsible for this escalation in prevalence remain unclear, and strategies for allergy prevention remain limited. OBJECTIVE: This article discusses current recommendations for allergy prevention and highlights new insights into allergic disease. DISCUSSION: History of allergic disease in a first degree relative is currently the only useful indicator for increased risk of developing allergic disease in a child. Prevention strategies should be directed to these high risk individuals. Currently, maternal dietary restriction during pregnancy or lactation and aeroallergen avoidance are not recommended. Breastfeeding is recommended, and where not possible or insufficient, a partially hydrolysed formula should be used in high risk infants. Introduction of solids should be delayed to 4-6 months of age. There is no evidence that delaying solids beyond this age is of benefit. There is currently insufficient evidence to recommend the addition of probiotics for allergy prevention. PMID- 18398515 TI - Allergy testing and referral in children. AB - BACKGROUND: Allergic diseases (asthma, atopic dermatitis, allergic rhinitis and food allergy) are the commonest chronic diseases of childhood. General practitioners commonly encounter children with allergic diseases and need to be aware of when referral to a paediatric allergist should be considered. An understanding of what diagnostic tests the allergist may use in confirming the diagnosis is also necessary. OBJECTIVE: This article discusses the criteria for referral to a specialist paediatric allergist and also details the tests that may be used by the allergist as part of the diagnostic work up. DISCUSSION: Management of allergic diseases requires accurate diagnosis and avoidance of offending allergens where possible. The diagnosis of an IgE mediated allergy requires both a history of symptoms on exposure to the allergen and detection of allergen specific IgE. The most commonly employed diagnostic methods in clinical allergy assessment are skin prick testing, RAST and clinical oral food challenge procedures. The use of alternative or unorthodox tests may provide misleading results and delay correct diagnosis and therefore should not be used. PMID- 18398516 TI - Allergic rhinitis - practical management strategies. AB - BACKGROUND: Allergic rhinitis is a common condition associated with significant effects on quality of life. Readily available treatments can improve outcomes in rhinitis as well as associated allergic diseases such as asthma. Yet allergic rhinitis remains underdiagnosed and undertreated. OBJECTIVE: This article outlines practical strategies and evidence based management of allergic rhinitis. DISCUSSION: Allergic and nonallergic rhinitis often co-exist. Thorough history of allergen exposure and its relationship to symptoms is vital for the ordering and interpretation of investigations and for management decisions. Some allergen avoidance measures may be ineffective and may cause an unnecessary burden. Demonstrated effective strategies are patient education, intranasal steroids and immunotherapy ('desensitisation'). General practitioners play a vital role in all three strategies, and in supporting patients and families to self manage what is often a chronic condition. PMID- 18398517 TI - Food allergy in adults. AB - BACKGROUND: There is a marked increase in the prevalence of food allergies. Food allergy can cause fatal anaphylaxis and the victims are most often adolescents and young adults. OBJECTIVE: This article focuses on IgE mediated food allergy and provides a review of the diagnostic and management strategies for food allergy, including a treatment algorithm for anaphylaxis. The role of the general practitioner in food allergy, when to refer to an allergist, and how to support patients with food allergies long term, including survival tips for patients with food allergy, are also discussed. DISCUSSION: The key management of food allergy is allergen avoidance informed by accurate allergy diagnosis. Inadvertent exposure to food triggers unfortunately does occur and patients need to be confident in prompt self management. Adrenaline must be given for all potentially life threatening food allergy reactions. Anaphylaxis action plans and optimal asthma control are also critical management objectives. PMID- 18398518 TI - Management of spinal cord injury in general practice - part 1. AB - BACKGROUND: Spinal cord injury (SCI) is a complex disability, often requiring specialised knowledge and expertise to manage multisystem impairments. This topic is often not included in undergraduate medical education and the general practitioner may understandably feel underprepared for managing a patient with SCI. OBJECTIVE: This two part article provides an overview of common complications associated with SCI. DISCUSSION: Common management issues in SCI include neurogenic bladder and bowel dysfunction, autonomic dysreflexia, sexuality, fertility, pressure areas, pain, spasticity, musculoskeletal disorders, neurological complications, general health and psychological wellbeing. Armed with key information, management of the patient with SCI can be systematically addressed in the general practice setting. PMID- 18398519 TI - In general practice, 'always expect the unexpected'. AB - CASE STUDY: Mr SF, aged 72 years, presented to a senior colleague complaining of a scalp sore which was failing to heal. The patient had injured his head while mustering cattle 4 years earlier. He consulted his local medical officer at that time and was reassured and sent on his way. Six weeks before presenting to our practice, Mr SF had split his head open again. Although he was not overly concerned about it at the time, it had been slow to heal and he had consulted a naturopath. The naturopath was packing the scalp sore with comfrey leaves and had advised Mr SF to eat curry to aid with its healing. He had been seeing this alternative practitioner each week for the preceding 6 weeks. Mr SF had become disillusioned with the poor results he was getting. At the behest of his wife he was seeking another opinion. PMID- 18398520 TI - Pathological hoarding. AB - BACKGROUND: Compulsive hoarding is defined, in most cases, as 'the inability to resist the urge to acquire possessions and to discard possessions'. Compulsive hoarding has an early age onset, chronic course and significant levels of comorbidity and morbidity. OBJECTIVE: This article outlines the essential features of compulsive hoarding occurring as a symptom factor of obsessive compulsive disorder (OCD) and alludes to hoarding co-occurring in a range of disorders other than OCD. A test screening for compulsive hoarding is provided. DISCUSSION: Compulsive hoarding occurs in illnesses other than OCD. Limited treatment studies have been completed only when compulsive hoarding is a symptom factor of OCD. These studies suggest that a pharmacological approach and multimodal cognitive behavioural therapy can achieve a moderate treatment outcome. The illness however is difficult to treat. PMID- 18398521 TI - Lessons from the TAPS study - errors relating to medical records. AB - The Threats to Australian Patient Safety (TAPS) Study collected 648 anonymous reports about threats to patient safety from a representative random sample of Australian general practitioners. These contained any events the GPs felt should not have happened and would not want to happen again, regardless of who was at fault or the outcome of the event. This series of articles presents clinical lessons resulting from the TAPS study. PMID- 18398522 TI - Is it insulin allergy? AB - 'I'm allergic to that insulin you gave me. The injections sting and leave marks that last more than a week'. When Joe pulls up his shirt you can see the marks. PMID- 18398523 TI - Supporting clinical teachers - a review of the literature. AB - BACKGROUND: Medical student teaching in the community has been shown to produce equivalent or improved learning outcomes compared to teaching based in the hospital setting. It is well recognised by Australian universities that there is a current shortage of clinical teachers, that these teachers are both under resourced and underfunded and that the majority of clinical teachers have no formal training in medical education. OBJECTIVE: A review was undertaken to determine what body of knowledge exists in the Australian literature regarding the educational support needs of general practitioners who teach medical students, and to highlight areas that require further research. DISCUSSION: Need for improved support was identified in all Australian articles that assessed GP support requirements. There is sufficient evidence to guide improved support for GP clinical teachers. However, funding for more support programs is needed, as is further research into the area. PMID- 18398524 TI - Doctors as patients. AB - Case histories are based on actual medical negligence claims or medicolegal referrals; however certain facts have been omitted or changed by the author to ensure the anonymity of the parties involved. All the medical boards and medical colleges recommend that medical practitioners have their own general practitioner. The aim of this article is to provide guidance for GPs about how to deal with the potential complexities of managing medical colleagues as patients. PMID- 18398525 TI - Colorectal cancer management - the role of the GP. AB - BACKGROUND: Colorectal cancer (CRC) contributes significantly to the morbidity and mortality of Australians. The introduction of the National Bowel Cancer Screening Program has focussed attention on the role of the general practitioner in CRC screening. However, their increasing role across the management spectrum of CRC remains poorly researched. OBJECTIVE: This article examines the published literature on the existing and potential roles of the GP in the detection and management of CRC. Evidence about the existing and potential role of the GP was found through a literature search on Medline, PubMed, epidemiological evidence and recent guidelines. DISCUSSION: The role of the GP in CRC management varies. Some GPs play many roles such as advocate, facilitator, supporter, educator and counsellor. The role of the GP is influenced by GP-specialist communication, practice location, and patient and GP factors. There is a potential to increase this role, especially in providing psychosocial support and counselling for both the patient and their family. PMID- 18398526 TI - Managing shoulder pain in general practice - assessment, imaging and referral. AB - BACKGROUND: A case review was carried out on 112 cases of shoulder pain, referred for specialist attention. The general practitioner referral letter offered a diagnosis in 40% (45/112) of cases with 89% (40/45) of these concurring with the diagnosis made by the orthopaedic surgeon. Shoulder ultrasound had been ordered for 95/112 (85%) patients with 71/112 (63%) being combined with plain radiology. OBJECTIVE: This article presents findings of a case review examining the management of shoulder pain in general practice. DISCUSSION: The assessment of shoulder presentation suggests a lack of confidence by the referring practitioner due to the reliance on diagnostic ultrasound. Such practice can be unnecessarily expensive and would benefit from the establishment of guidelines for what imaging best suits the presenting complaint. PMID- 18398527 TI - Too many of us have gone early - priorities in heart health education for Aboriginal people. AB - Aboriginal people have a higher burden of cardiovascular risk factors and heart disease and poorer outcomes after heart attacks when compared with other Australians. Indigenous status is also a risk factor for delayed response to heart attack symptoms. A community DVD about preventing and managing heart disease was made at the Aboriginal Medical Service Western Sydney in 2005. The aim of this article is to reflect on the process of making the DVD as a community driven health promotion activity and to explore questions raised and insights gained about heart health education for Aboriginal people in the context of the existing literature. The importance of education about heart attack symptom recognition and prompt hospital presentation, as well as risk factor management, by general practitioners and other health practitioners working with Aboriginal people, is highlighted. PMID- 18398528 TI - Telephone based asthma management - financial and individual benefits. AB - METHODS: The authors calculated the cost of delivering telephone based asthma management compared to usual care. Self efficacy and quality of life were measured at recruitment, 6 months and 12 months. Participants were invited to comment on their experience of the telephone based intervention. RESULTS: The intervention was well accepted. At 12 months there was a clinically important improvement in mean quality of life in the intervention group not seen in the control group. Telephone based management costs were offset by reductions in the number of readmissions in the intervention group. DISCUSSION: Telephone based asthma management offers a well accepted, low cost yet potentially effective means of delivering asthma care. PMID- 18398529 TI - General practice registrars and research - attitudes toward participation. AB - BACKGROUND: Early exposure of general practitioners to research is recommended to increase family medicine research capacity. However, vocational training programs encounter difficulties in engaging general practice registrars in research projects. The authors investigated registrars' opinions of research and their participation in research in daily practice. METHODS: Sixty-seven Dutch general practice registrars participated in a trial concerning patients with emotional symptoms. The authors assessed the registrars' participation and opinions through observation and a questionnaire. RESULTS: Response rate was 82%. Registrars recruited 208 patients. The participants liked learning a new skill and participating in research. Obligatory participation, lack of time and difficulties with patient recruitment were important barriers to participation. DISCUSSION: Registrars report that participation in research during vocational training is interesting but that it should not be compulsory, and that they prefer to choose their own research subjects. The authors recommend implementing an attractive research program during vocational training. PMID- 18398530 TI - Coordinated anticoagulation management in a rural setting. AB - BACKGROUND: Oral anticoagulation management is difficult in rural settings because of reduced patient access to pathology testing and medical management. Previous research reports the effectiveness of coordinated anticoagulation management incorporating education, point of care international normalised ratio (INR) testing, patient self care models, protocols and use of specially trained personnel. This article presents findings on the assessment of a Victorian rural program using a modified anticoagulation clinic and other strategies to improve anticoagulation management. METHODS: This program assessed multiple strategies including comprehensive patient education, protocols and point of care INR testing. These were implemented in a rural hospital and rural general practices. Specific measures for evaluation were time in the therapeutic INR range and complication rate. RESULTS: Time in the therapeutic INR range was 69% for the standard range (2.0-3.0) and 81% using an expanded range (1.8-3.0). The anticoagulation related complication rate was 0.03 per patient year (95% CI: 0.01, 0.06). International normalised ratio testing every 14 days resulted in 78% of time spent in therapeutic range. DISCUSSION: The strategies employed in the study increased time spent in therapeutic range and reduced anticoagulation related complications. PMID- 18398531 TI - Using computer based templates for chronic disease management. AB - BACKGROUND: General Practice Management Plans (GPMPs) and Team Care Arrangements (TCAs) were introduced in 2005 to promote better chronic disease management and to provide funding to support general practitioner involvement in care planning. To develop these plans, GPs complete forms documenting goals and strategies using computer based templates. OBJECTIVE: This article evaluates GP use of computer based templates for GPMPs and TCAs, their views about using them, and community health service (CHS) staff attitudes to the role of TCAs in coordination of care. METHOD: A qualitative interview based study of 31 GPs from solo and group practices and 15 service providers from community health centres (n=46). DISCUSSION: Most GPs interviewed used templates in claiming the GPMP and TCA items. Reasons GPs chose not to use the items included time constraints and uncertainty about the process. Community health service staff identified difficulties with TCAs and care coordination. This study suggests that templates assist GPs in claiming for GPMP and TCA Medicare Benefits Schedule item numbers but do not in themselves facilitate care coordination. To improve care coordination between general practice and other agencies, communication regarding TCAs must be improved. PMID- 18398532 TI - Light-emitting iridium complexes with tridentate ligands. AB - Once the Cinderella amongst the Platinum Group Metals at the Photochemistry Ball, iridium has become of intense interest since the beginning of the decade. Complexes of iridium(III) can be prepared that are highly luminescent, with emission wavelengths tuneable over the whole of the visible region. Whilst most studies have focused on tris-bidentate complexes, a rich and varied chemistry is also possible using tridentate ligands. In this review, we discuss the synthesis and excited-state properties of such complexes, exploring in particular how the number of cyclometallating carbon atoms in the coordination sphere of the metal ion influences the luminescence. Moving from [IrN6]3+ to [IrN3X3] coordination via [IrN5X]2+ and cis/trans-[IrN4X2]+ complexes, where N is a heterocyclic nitrogen and X is an anionic ligand or cyclometallated carbon, a whole range of luminescence efficiencies are encountered, ranging from the barely detectable to room temperature quantum yields approaching unity. We consider the extent to which these profound differences, arising as a result of subtle changes in molecular structure, can be rationalised in terms of the nature of the frontier orbitals. PMID- 18398533 TI - Self-assembly reactions with a bis(benzene-o-dithiolato) ligand. AB - Reaction of the 1,5-diamidonaphthalene bridged bis(benzene-o-dithiol) ligand H4- with Ti4+ leads to the unexpected formation of a dinuclear meso-complex anion [Ti2(B)3](4-), while the catechol analog H(4-) has been reported to yield the tetrahedral [M4(A)6](8-) cluster. PMID- 18398534 TI - 3D coordination metal-organic frameworks of octacyanometalate bridging between Cu4 magnetic units. AB - Magnetic Cu4 clusters with S = 2 are bridged by octacyanometaltate(IV) to form two 3D cluster arrays of metal-organic frameworks. Magnetic investigation shows the ferromagnetic coupling between Cu(II) ions and very weak antiferromagnetic interaction between clusters. PMID- 18398535 TI - Cellular confocal fluorescence studies and cytotoxic activity of new Zn(II) bis(thiosemicarbazonato) complexes. AB - We report the synthesis and characterisation of new, highly fluorescent, zinc complexes of bis(thiosemicarbazone) ligands incorporating extended aromatic backbones which are cytotoxic at levels comparable to cisplatin; cellular fluorescence imaging studies suggest these cause cell death by disruption of mitochondria. PMID- 18398536 TI - En route to zirconium hydrazides(2-). AB - The structures and properties of a series of new zirconium hydrazido(1-) complexes and the possibility of converting them to the respective hydrazido(2-) species are reported. Reaction of complex [Zr(N2TBSNpy)Cl2] (1) with the monolithiated hydrazide LiNHNMe2 gave the hydrazido(1-) complex [Zr(N2TBSNpy)(NHNMe2)Cl] (2) which exists as two isomeric forms (2a and 2b) in solution. All attempts to convert a mixture of 2a and 2b to the respective hydrazido(2-) compound by reaction with the bulky base lithium hexamethyldisilazide or via the alkyl/hydrazido(1-) complex [Zr(N2TBSNpy)(CH2SiMe3)(NHNMe2)] (3) and subsequent thermal alkane elimination failed. Reaction of 1 with LiHNNPhMe gave a mixture of stereoisomers of [Zr(N2TBSNpy)(NHNMePh)Cl] (4a and 4b), in which the hydrazido unit is end-on bound in solution and eta 2-bonded in the solid state. Reaction of this mixture with lithium hexamethyldisilazide in the presence of pyridine selectively yielded the hydrazido(2-) complex [Zr(N2TBSNpy)(NNPhMe)(py)] (5) which aggregated upon attempts to isolate it. Reaction of the insoluble precipitate with 4 dimethylaminopyridine (dmap) selectively gave the corresponding hydrazido(2-) complex [Zr(N2TBSNpy)(NNPhMe)(dmap)] (6), which could be obtained in a one-pot reaction directly from 1 and which was analytically and spectroscopically fully characterized. It appears that the isolation of stable hydrazido(2-) complexes of zirconium depends on the type of substituents at the N beta atom as well as the co-ligands coordinated to the metal centre. PMID- 18398537 TI - Stable Lewis acid chelate of a bis(imido) tungsten compound and implications for alpha-olefin dimerisation catalysis: a DFT study. AB - Dimerisation of alpha-olefins by a catalyst system comprising of a bis(imido) tungsten compound and a Lewis acid component has been investigated by density functional theory employing a catalyst model that closely mimics the real system together with ethylene chosen as a prototypical substrate. This study disclosed that the Lewis acid preferably forms four-membered chelates by bridging across tungsten-imido linkages. Complexation at one imido group is predicted as the favourable, stable mode of Lewis acid association onto a bis(imido) tungsten(VI) compound. The computational analysis revealed that the cocatalyst affects the energetics of the several steps to a different extent. Formation of the five membered tungstanacycle and the further growth is moderately influenced, this is in contrast to its profound influence on metallacycle degradation. AlClMe2 chelation of both tungsten-imido groups is seen as being most effective in smoothing the energetics of the dimer-generating route. The predicted energy profile of the most accessible pathway, however, is at odds with observed catalytic abilities. This study indicates that the removal of one imido ligand by the Lewis acid is a viable process, thereby suggesting that mono(imido) tungsten compounds are effective in dimerisation catalysis of the reported catalyst system. PMID- 18398538 TI - Selenium-bridged diiron hexacarbonyl complexes as biomimetic models for the active site of Fe-Fe hydrogenases. AB - Three N-substituted selenium-bridged diiron complexes [{(mu SeCH2)2NC6H4R}Fe2(CO)6] (R = 4-NO2, 7; R = H, 8; R = 4-CH3, 9) were firstly prepared as biomimetic models for the Fe-Fe hydrogenases active site. Models could be generated by the convergent reaction of [(mu-HSe)2Fe2(CO)6] (6) with N,N bis(hydroxymethyl)-4-nitroaniline (1), N,N-bis(hydroxymethyl)aniline (2), and N,N bis(hydroxymethyl)-4-methylaniline (3) in 46-52% yields. All the new complexes were characterized by IR, 1H and 13C NMR and HRMS spectra and their molecular structures were determined by single-crystal X-ray analysis. The redox properties of and their dithiolate analogues [{(mu-SCH2)2NC6H4R}Fe2(CO)6] (R = 4-NO2, 7s; R = H, 8s; R = 4-CH3, 9s ) were evaluated by cyclic voltammograms. The electrochemical proton reduction by and were investigated in the presence of p toluenesulfonic acid (HOTs) to evaluate the influence of changing the coordinating S atoms of the bridging ligands to Se atoms on the electrocatalytic activity for proton reduction. PMID- 18398539 TI - Synthesis, characterization and electrochemistry of 4'-functionalized 2,2':6',2'' terpyridine ruthenium(II) complexes and their biological activity. AB - The synthesis and characterization of Ru(II) terpyridine complexes derived from 4'-functionalized 2,2':6',2''-terpyridine ligands by a multi step procedure have been described. The complexes are redox-active, showing both metal-centred (oxidation) and ligand-centred (reduction) processes. The antibacterial and antifungal activity of the synthesized ruthenium(II) complexes [Ru(attpy)2](PF6)2 (attpy = 4'-(4-acryloyloxymethylphenyl)-2,2':6',2''-terpyridine); [Ru(mttpy)2](PF6)2 (mttpy = 4'-(4-methacryloyloxymethylphenyl)-2,2':6',2''- terpyridine); [Ru(mttpy)(MeOPhttpy)](PF6)2 (MeOPhttpy = 4'-(4-methoxyphenyl) 2,2':6',2''-terpyridine); and [Ru(mttpy)(ttpy)](PF6)2 (ttpy = 4'-(4-methylphenyl) 2,2':6',2''-terpyridine) were tested against four human pathogens (Proteus vulgaris, Proteus mirabilis, Pseudomonas aeruginosa and Escherichia coli) and five plant pathogens (Curvularia lunata, Fusarium oxysporum, Fusarium udum, Macrophomina phaseolina and Rhizoctonia solani) by the well diffusion method and MIC values of the complexes are reported. A biological study of the complexes indicated that the complexes [Ru(mttpy)2](PF6)2 and [Ru(mttpy)(MeOPhttpy)](PF6)2 exhibit very good activity against most of the test pathogens and their activity is better than those of some of the commercially available antibiotics like tetracycline and the fungicide carbendazim. PMID- 18398540 TI - Eta 1 and eta 2 Coordination of 1-amino-closo-dodecaborate. AB - The reaction of the sodium salt of 1-amino-closo-dodecaborate [Na]2[NH2-B12H11] ([Na]2[1]) with [Au(PPh3)Cl] and [Ni(THF)2(Br)2] led to eta 1(N) coordination of 1in [Na][Au(PPh3)(NH2-B12H11)] (2) and [Na]6[Ni(NH2-B12H11)4] (3), respectively. Furthermore, eta 2(N,BH) coordination of was found in [MePPh3][Rh(PPh3)2(NH2 B12H11)] (4), which was synthesized by the reaction of [MePPh3][Na][1] with [Rh(PPh3)3Cl]. All compounds were characterized by single crystal X-ray diffraction and heteronuclear NMR spectroscopy. PMID- 18398541 TI - Electrochemical fabrication and characterization of thin films of redox-active molecular wires based on extended Rh-Rh bonded chains. AB - An original electrochemical synthesis of {[Rh4(mu-OOCCH3)4(phen)4]2+}n (1) molecular wire films from a solution of binuclear bridged Rh complexes [Rh2(mu OOCCH3)2(phen)2(X)2](Y)2 (X = H2O, Y = BF4(-) (2a) and X = CH3CN, Y = BF4(-) (2b)) in MeCN electrolyte is reported. UV-vis spectroscopy and quartz crystal microbalance electrochemical coupled techniques have been used to demonstrate the electrosynthesis process. The resulting polymetallic compound has been characterized on the basis of its physicochemical properties, which have been compared with those of a chemically synthesized sample. Furthermore, according to EPR, 1H NMR and electrochemical behaviour, the mechanism of the oxidation of this polymetallic wire, containing mixed valent rhodium centers and alternatively acetate bridged Rh-Rh bonds, has been investigated in detail. PMID- 18398542 TI - Non-covalent DNA binding and cytotoxicity of certain mixed-ligand ruthenium(II) complexes of 2,2'-dipyridylamine and diimines. AB - A series of mixed ligand ruthenium(II) complexes [Ru(Hdpa)2(diimine)](ClO4)2, 1-5 where Hdpa is 2,2'-dipyridylamine and diimine is 1,10-phenanthroline (phen) and a modified/extended 1,10-phenanthroline such as, 5,6-dimethyl-1,10-phenanthroline (5,6-dmp), dipyrido[3,2-d:2',3'-f]quinoxaline (dpq), 5-methyldipyrido[3,2-d:2',3' f]quinoxaline (mdpq) and dipyrido[3,2-a:2',3'-c]phenazine (dppz) have been isolated and characterized by analytical and spectral methods. The complex [Ru(Hdpa)2(phen)](PF6)2 1 has been structurally characterized and the coordination geometry around Ru(II) in it is described as distorted octahedral. 1H NMR spectral data reveal that 1-5 should have a C2 symmetry lying on the diimine plane due to the rapid flapping of the coordinated Hdpa ligands. The interaction of the complexes with calf thymus (CT) DNA has been explored by using absorption and emission spectral and viscometry and electrochemical techniques and the mode of DNA binding of the complexes has been proposed. The DNA binding affinity of the complexes decreases with decrease in number of planar aromatic rings in the co-ligand supporting the intercalation of the diimine co-ligands in between the DNA base pairs. Circular dichroic spectral studies reveal that the complexes 3-5 exhibit induced circular dichroism upon binding to CT DNA. Interestingly, upon interaction with CT DNA all the complexes show an increase in anodic current in the cyclic voltammograms suggesting that they are involved in electrocatalytic guanine oxidation. Interestingly, of all the complexes, only 5 alters the DNA superhelicity upon binding with supercoiled pBR322 DNA, which is consistent with its higher DNA binding affinity. Further, the cytotoxicities of the complexes against human cervical epidermoid carcinoma cell line (ME180) have been examined. Interestingly, 5 exhibits a cytotoxicity against ME180 higher than other complexes with potency approximately 8 times more than cisplatin for 24 h incubation but 4 times lower than cisplatin for 48 h incubation. PMID- 18398543 TI - New atropisomeric N-N ligands for CO/vinyl arene copolymerization reaction. AB - New atropisomeric bidentate bipyridine-based ligands (3,3'-(ethylenedioxy)-2,2' bipyridine 4; 3,3'-(propylenedioxy)-2,2'-bipyridine 4; 3,3'-(butylenedioxy)-2,2' bipyridine 5) containing a bridge between the 3,3' positions of the aromatic rings have been prepared. Together with the previously reported analogous ligands ((R)-3,3'-(1-methylethylenedioxy)-2,2'-bipyridine) 1and ((S,S)-3,3'-(1,2 dimethylethylenedioxy)-2,2'-bipyridine) 2, they were used to synthesize the corresponding bis-chelated dicationic complexes [Pd(N-N)2][PF6]2. Crystal structures and comparison of the data obtained by X-ray analysis on four of these complexes is reported. These palladium compounds were used as precatalysts in the CO/styrene and CO/4-Me-styrene copolymerization reactions, where they showed that small variations in the ligand backbone remarkably affects the productivity of the catalytic system. PMID- 18398544 TI - Syntheses, structures and magnetism of linear tri- and tetra-copper chains containing anions of N,N'-bis(pyrimidine-2-yl)formamidine. AB - Reaction of Kpmf (pmf(-) = anion of N,N'-bis(pyrimidyl-2-yl)formamidine, Hpmf) with divalent copper salt CuX2 afforded the linear trinuclear complexes of the type [Cu3(pmf)4](X)2 (X = BF4, 1; NO3, 2; ClO4, 3), while reaction of Kpmf with monovalent copper salt CuX gave the linear tetranuclear complexes of the type Cu4(pmf)4X2 (X = Cl, 4; Br, 5). The copper atoms of complexes 1-5 are helically bridged by four pmf(-) ligands, resulting in three different coordination modes for the pmf(-) ligands. In complexes 1-3, one pmf(-) ligand adopts a new coordination mode with the two amine nitrogen atoms chelating to the central copper atom, while the other three feature chelation by one pyrimidyl and one adjacent amine nitrogen atoms. The Cu(II)...Cu(II) distances are 2.729(2) and 2.825(2) A for 1, 2.762(1) and 2.832(1) A for 2 and 2.732(1) and 2.827(1) A for 3. In complexes 4 and 5, the pmf(-) ligands are coordinated to the copper atoms in tetradentate fashion with each nitrogen atom coordinating to one Cu atom. The Cu...Cu distances are 2.580(1) and 2.549(1) A for 4 and 2.582(1) and 2.561(1) A for 5. Antiferromagnetic interactions between the copper ions are observed with calculated g and J values of 2.03(1) and -188(2) cm(-1) for 1, 2.09(1) and 268(3) cm(-1) for 2, and 2.09(1) and -486(2) cm(-1) for 5. By comparing the magnetic data it can be shown that the bonding mode of the pmf(-) ligand is one of the important factors in determining the strengths of the Cu...Cu interactions in linear trinuclear and tetranuclear copper complexes. PMID- 18398545 TI - Ruthenium complexes with tridentate PNX (X = O, S) donor ligands. AB - The ligands, PhPNXMe (1), PhPNXPh (2), and PhPNSMe (3), (PhPNX = 2-Ph2P C6H4CH[double bond, length as m-dash]NC6H4X-2; X = O, S) have been prepared. A range of new ruthenium complexes were synthesised using these and related ligands, namely: [{RuCl(PhPNO)}2Cl] (4), [Ru(PhPNO)2] (5), [RuCl(PhPNXR)(PPh3)]BPh4 [X = O, R = Me (6); X = O, R = Ph (7); X = S, R = Me (8)], [{RuCl(PhPNX'R)}2Cl]X [X' = O, R = Me, X = Cl(-) (9); X' = S, R = Me, X = BPh4(-) or PF6(-) (10)], and [RuCl(PhPNO-eta 6C6H5)]BPh4 (11). The catalytic activity of these complexes with respect to the hydrosilyation of acetophenone and the hydrogenation of styrene has been investigated, giving an insight into the requirements for an active complex in these reactions. PMID- 18398547 TI - [On evidence-based public health]. PMID- 18398546 TI - Gd-complexes of DTPA-bis(amide) conjugates of tranexamic acid and its esters with high relaxivity and stability for magnetic resonance imaging. AB - The synthesis and the characterization of a series of DTPA-bis(amide) conjugates of tranexamic acid (L1), its esters (L2-L6), and their Gd(III) complexes of the type [Gd(L)(H2O)].nH2O (L = L1-L6) are described. Except for the case of , all Gd complexes exhibit greatly enhanced R1 relaxivity. Highest R1 reaches up to 12.9 mM(-1) s(-1) for [Gd(L2)(H2O)]. Such high relaxivity is reflected in the intensity enhancement of the in vivo MRI study on H-ras transgenic mice bearing hepatic tumor when employing [Gd(L2)(H2O)] as an MRI contrast agent. Thermodynamic stability constants, conditional stability constants, and the pM values demonstrate higher stability of [Gd(L)(H2O)].nH2O (L =L1-L6) than Omniscan under physiological conditions. The MTT assay performed on these complexes reveals cytotoxicity as low as that for Omniscan in the concentration range required to obtain intensity enhancement in the in vivo MRI study. PMID- 18398548 TI - [Evidence Based Public Health: resources on effectiveness of community interventions]. AB - The evaluation of interventions in Public Health is a key element through the process of developing health policies, but it is not free of controversy. For doing this purpose it is essential the use of research outcomes, although there are sticking points related to the traditional approach of Evidence Based Medicine, dominated by the randomized clinical trial as the gold standard. Not always it is possible to develop randomized and controlled studies in Public Health (sometimes due to ethical limitations, or because of the technical impossibility for performing the trial or because conceptual incompatibility) and the interventions are mostly multifaceted, therefore, the interpretation of the results is a complex task. In other hand, the usual criteria for research appraisal underestimates systematically the observational studies which, frequently, are the indicated in Public Health scenarios. Nevertheless, a great advance has been implemented with the generation of strategies as TREND (Transparent Reporting of Evaluations with Non randomized Designs), as well as other instruments like STROBE (STrenghtening the Reporting of OBservational studies in Epidemiology) or MOOSE (Meta-analysis Of Observational Studies in Epidemiology). But regardless of the existence of more or less consolidated critical appraisal tools, we all need a solvent and rigorous way of knowing the outcomes of Public Health interventions. This would make more dynamic the review, design or planning phases, and it would contribute to facilitate the decision making process when a well grounded knowledge be available. In this paper all the methodological process about searching evidence in Public Health interventions is reviewed, as well as the main sources providing this information, in order to facilitate this task to the Public health professionals. PMID- 18398549 TI - [Ethics guidelines for the creation and use of registries for biomedical research purposes]. AB - The clinical information stored in registries and records of different types is a fundamental tool for biomedical research. Up until just a few years ago, hardly any limitations existed on the creation and use of epidemiological registries or the use of information from pre-existing records for research purposes. This situation has changed substantially due mainly to the growing importance current laws place upon the safeguarding of the privacy and confidentiality of personal data. Although the legal framework is already quite explicit, a certain degree of leeway exists for ethical debate and prudence advice for the purpose of conducting valid, useful research with this information which will also respect the rights of the subjects and the laws in force. These guidelines deal with those aspects which have been considered relevant from an ethical standpoint in the handling of records and registries for research-related purposes, including not only the use but also the creation proper of the registries. A total of twenty-four recommendations are provided, grouped into ten sections: warranting of the creation of registry, organization and definition of responsibilities, scientific validity of the research project, ethical requirements of the collections of anonymous and anonymized data, ethical requirements of the registries including personal data, uses of medical records for research purposes, use of historical records of deceased individuals, contact with the research subjects, notification of results and review by a Research Ethics Committee. PMID- 18398550 TI - [Epidemiology of falls in the elderly in Spain: a systematic review, 2007]. AB - Falls in the elderly are a serious problem that results in large health care expenditures. To prevent them, it has been shown that the key is an epidemiologic knowledge of the target population. The purpose of this systematic review was to identify the incidence, risk factors and consequences of falls in the Spanish population, by analyzing the methodological quality of studies that provide these data. Two independent authors identified 54 studies through manual and electronic means (MEDLINE, Dialnet, Tesis en Red, TESEO y CSIC [ICYT, IME, ISOC]). A total of 13 studies performed in Spain with non-hospitalized elderly individuals older than 64 years of age were selected and assessed for their methodological quality. We found heterogeneity in the characteristics and quality of the studies, and a general inadequacy of data analyses. The risk factors and consequences must be viewed with caution, since in most of the studies a causal inference cannot be made. We showed that the current fall rates are at the same level as those of the first epidemiologic study published 15 years ago. We conclude that Spain has a high incidence of falls and needs studies on risk factors directed toward cause and effect in the community and comparisons among nursing homes. Finally, the physical, psychosocial and economic consequences must be investigated more thoroughly. PMID- 18398551 TI - [Cycles of improvement of the hypertensive patient's care: risk-cycle study]. AB - BACKGROUND: Frequently we found deficiencies in the management of cardiovascular risk factors on hypertensive patients in primary care. OBJECTIVE: This study was to evaluate the effect of a quality improvement intervention in the care of cardiovascular risk factors in hypertensive patients. METHODS: Quality assurance study. Two health centres with fourteen family doctors. One centre (seven doctors) was assigned to receive a quality improvement intervention while the other centre was assigned the control group. 482 hypertensive patients were in the study group (64% females, mean age 61,4 years (SD 6,8)), and 360 were in the control group (63% females, mean age 60,7 (SD 7,4)). Quality improvement circles consisting of audit, feedback, training sessions, and guidelines discussion. Process criteria of hypertension control, blood pressure, weight, lipids, smoking, cardiovascular risk and antihypertensive drugs used were measurement before intervention and again one year later. RESULTS: The mean improvement in process criteria after the intervention was 5,3 percent points (CI95%:3,7-6,9). Systolic blood pressure was decreased by 3,5 mmHg(IC95%:1,6-5,3) and Diastolic blood pressure by 2,5 mmHg(IC95%:1,3-3,8). Adequate BP control was significantly increased in the intervention group (29,1% to 40,9%;p<0,01), while no effect was achieved in the control group. Absolute cardiovascular risk decreased (15,86 to14,34%, p<0.01) in the study group, with no changes in the control group. Absolute risk decreases 2,07(IC95%:1,21-2,93) and relative risk 0,25 (IC95%:0,14 0,35) percent points. CONCLUSION: The quality intervention was effective in improving the quality care process and decreased blood pressure and absolute and relative cardiovascular risk. PMID- 18398552 TI - [Evaluation of a health care programme with nursing tutor in a hospital traumatological facility in a hospital, Granada, Spain, 2004]. AB - BACKGROUND: The current trend in patient care is towards a more humanized and higher-quality healthcare. The objective of the present study was to evaluate the results of implementing a programme of healthcare with a reference nurse who welcomes the patient at hospital admission, visits regularly during hospital stay, and resolves doubts and problems. METHODS: Quasi-experimental study (149 cases and 454 non-tutored controls) in patients admitted for scheduled trauma surgery at the Virgen de las Nieves University Hospital, Granada. Sociodemographic and clinical data were gathered at admission. At 15 days after discharge, patients were administered with an interviewer-administered Zung score and an ad hoc questionnaire on satisfaction with different healthcare aspects during hospital stay and understanding of information received. Bivariate analyses and linear and multivariate logistic regression analyses were performed with a study of confounding variables. RESULTS: For the outcome variable anxiety level non-tutored patients had a B coefficient of 2.64 (p<0,01) in the multivariate linear regression analysis controlling for the other variables in the final model: sex, presence of informal career, professional activity, days of hospital stay and understanding of health information. For the outcome variable inadequate understanding information non-tutored patients showed an odds ratio of 3.48 in the multivariate analysis controlling for educational level and presence of informal career. Satisfaction with he friendliness of health care personnel and with the hospital setting did not significantly difference, although the percentage of dissatisfied patients was higher in the non-tutored group: 15% vs 11% (p= 0.34) and 18% vs 12 % (p= 0.11) respectively. CONCLUSIONS: The implemented continued care programme proved effective for these patients, increasing their understanding of information received and reducing anxiety levels. PMID- 18398553 TI - [Outbreak of streptococcal toxic shock syndrome in a day care center in Cantabria, Spain, 2006]. AB - BACKGROUND: Beta hemolytic group A streptococcus only exceptionally produces aggressive disease with high lethality. Even more uncommon is the occurrence of an outbreak. In Spain, no outbreak in child care center has been previously described. METHODS: Descriptive study of an outbreak of streptococcal toxic shock syndrome (3 cases, one lethal) in a child care center, which motivated the health care intervention with chemoprophylaxis, the closure of the child care center and the study of contacts. We analyzed the determinants of infection in the invasive and non-invasive cases, and the results of the pharyngeal culture of contacts. RESULTS: We identified 3 invasive and 14 non-invasive cases between 40 children attending the child care center (attack rate 42.5%). We studied 19 possible determinants of the infection, finding only an association with being over the age of 24 months and the assistance to the handouts classroom (that of the oldest children). It was not associated with chickenpox. All children attending the child care center, its staff (4 women) and 258 contacts were microbiologically investigated. In 12 children the emm 4 strain was isolated, including 2 of 3 cases with invasive disease. In 13 of 258 contacts other strains of beta hemolytic group A streptococcus were isolated, but in none of them the strain responsible of the outbreak was found. Azytromicin chemoprophylaxis was implemented for all children and contacts, and in those with a positive isolation, the culture was repeated until negative. CONCLUSIONS: The invasive strain circulated only in the child care center. Azytromicin chemoprophylaxis eradicated effectively the infection. PMID- 18398554 TI - [Temporary distribution of reports and murders because of partner violence during the period 1998-2006, Spain]. AB - BACKGROUND: Intimate Partner Violence (IPV) has received special attention in the spanish political agenda. Despite the importance of developed interventions, empirical evidence about their effectiveness is scarce. The aim of this study is to explore the temporary distribution of Intimate Partner Violence (IPV) reports and murders and to identify possible changes in the risk of dying and reporting by IPV from the Spanish law against gender violence of December of 2004. METHODS: We performed a descriptive study based on statistics of The Woman's Institute and The Divorced and Separated Women Federation. CALCULATIONS: IPV mortality and reports rates by years and periods around the law (1998-2004 vs. 2005-2006); Poisson Regression; and, Epidemic index by months -Ratio between the actual number of IPV murders and reports in a given month and the median number of cases in the same month in the five preceding years- and underlying epidemic index annual average of the scores of the epidemic index of reports and murders. RESULTS: The epidemic index trends permit us to observe that the problem has decreased since 2005. Nevertheless, the possibility of reporting IPV between 2005 and 2006 is 1.6 times upper that between 1998 and 2004. The risk of dying by this cause does not show statistically significant changes. CONCLUSION: The possibility of reporting IPV has been increased in Spain throughout the time. However, the risk of dying stays. Although still it is soon for the evaluation of the impact of the law in this epidemic, we could say that its effectiveness for the reduction of IPV mortality seems limited. PMID- 18398555 TI - [Social and economic impact of chicken pox vaccine at 15 months of age. Castile and Leon, Spain, 2004]. AB - BACKGROUND: Chicken pox is a mainly childhood contagious disease caused by the Varicella Zoster Virus which gives rise to major healthcare and social costs. In 2005, Castile and Leon added chicken pox vaccine injections to its childhood vaccination schedule for eleven year-olds subject to coming down with this disease. This strategy does not modify the major mobility generated thereby at younger ages. This study is aimed at evaluating the profitability of systematic vaccination for chicken pox in infants 15 months of age in Castile and Leon. METHODS: An economic cost-benefit evaluation has been set out by jeans of a decision-making tree. A fictitious cohort of 100,000 children in Castile and Leon having reached 15 months of age in 2004 is studied, to whom the chicken pox vaccine would be administered in conjunction with the mumps, measles, rubella vaccines. This study is approached from the social standpoint. The time horizon selected was that of up until the study cohort was to reach 15 years of age, applying a 3% discount rate. A sensitivity analysis was made for evaluating the uncertainty of some variables... RESULTS: The cost-benefit ratio of adding this vaccine to the childhood vaccination schedule amounts to 1.23. CONCLUSIONS: From the social standpoint, administering chicken pox vaccine in conjunction with the mumps, measles, rubella vaccines show itself to be profitable. The profitability is modified both if a second dose of vaccine is added as well as if only the direct healthcare costs are analyzed. PMID- 18398556 TI - [Aggregation of cases of legionella pneumonia in workers related to the ceramic industry in Castellon, Spain, 2006]. AB - BACKGROUND: Cases of legionellosis sometimes arise in the form of aggregations which are difficult to interpret. This study provides an analysis of these groupings having arisen one summer in Castellon. METHODS: Microbiological, clinical and epidemiological field methods were employed. RESULTS: Within a few days, there were five (5) cases of legionella pneumonia in workers related to various ceramic industries within an area where a large number of these plants are located. Two constituted a minor outbreak. By means of molecular biology techniques performed on the strains, the common origins of three of the cases were ruled out. CONCLUSIONS: It is debated whether this episode of an outbreak having arisen within the context of a broader-ranging cluster of cases of a multi focal origin was a chance event or whether it was the manifestation of a risk related to this industrial activity which might repeat itself. PMID- 18398557 TI - Shattered Shangri-la: differences in depressive and anxiety symptoms in students born in Tibet compared to Tibetan students born in exile. AB - OBJECTIVE: As a result of ongoing political tensions within Tibetan regions of the People's Republic of China, several thousand Tibetans escape across the Himalayas every year to seek refuge in India and Nepal. Prior studies have found a high prevalence of depressive and anxiety symptoms in these refugees, many of whom are young and have been exposed to significant trauma. However, it is not known whether depressive and anxiety symptoms are more prevalent in these refugees than in ethnic Tibetans born and raised in the relative political and social stability of exile communities in North India and Nepal. METHODS: We conducted a cross-sectional survey of a convenience sample of 319 students attending school at the Tibetan Children's Villages in Northern India to test the a priori hypothesis that adolescents and young adults who escaped from Tibet to India would demonstrate increased depressive and anxiety symptoms when compared to ethnic Tibetans born and raised in exile. The Hopkins Symptom Checklist-25 (HSCL-25) was used to measure depressive and anxiety symptoms. In addition, demographic information on age, sex, country of birth and frequency of family contact was collected. RESULTS: Students born in Tibet had higher mean HSCL-25 depressive and anxiety symptom scores than did ethnic Tibetans born in exile. Female students demonstrated higher depressive and anxiety scores, as did those with limited contact with immediate family. After adjusting for sex, age and frequency of family contact, being born in Tibet was associated with increased HSCL-25 depressive and anxiety symptom scores (depression: F[2, 316] = 29.96, P < 0.0001; anxiety: F[4, 316] = 43.57, P < 0.0001). CONCLUSIONS: The experience of being raised in Tibet and escaping to India appears to be a risk factor for increased depressive and anxiety symptoms when compared to being born and raised within an exile community in India or Nepal. PMID- 18398558 TI - The prevalence and socio-demographic correlations of depression, anxiety and stress among a group of university students. AB - BACKGROUND: The mental health of university students is an area of increasing concern worldwide. The objective of this study is to examine the prevalence of depression, anxiety and stress among a group of Turkish university students. METHODS: Depression Anxiety and Stress Scale (DASS-42) completed anonymously in the students' respective classrooms by 1,617 students. RESULTS: Depression, anxiety and stress levels of moderate severity or above were found in 27.1, 47.1 and 27% of our respondents, respectively. Anxiety and stress scores were higher among female students. First- and second-year students had higher depression, anxiety and stress scores than the others. Students who were satisfied with their education had lower depression, anxiety and stress scores than those who were not satisfied. CONCLUSIONS: The high prevalence of depression, anxiety and stress symptoms among university students is alarming. This shows the need for primary and secondary prevention measures, with the development of adequate and appropriate support services for this group. PMID- 18398559 TI - Mental health of migrant workers in China: prevalence and correlates. AB - BACKGROUND: This study aimed to examine the prevalence and the socio-demographic correlates of mental health of migrant workers in Shanghai China. METHODS: A total of 475 migrant workers from four major districts in Shanghai were recruited through a survey design with a multistage cluster [corrected] sampling. Male and female migrant workers were identified as mentally healthy or unhealthy using the brief symptom inventory. Socio-demographic characteristics and migration stress were explored as correlates of the mental health of the migrant workers. RESULTS: A total of 73 migrant workers could be classified as mentally unhealthy (25% for men and 6% for women). Male migrant workers who were married (OR 6.16, 95% CI 1.83-20.70), manual laborers (OR 1.56, 95% CI 0.97-2.51), and experienced more stress in "financial and employment-related difficulties" (OR 2.75, 95% CI 1.47 5.14) and "interpersonal tensions and conflicts" (OR 4.18, 95% CI 1.55-11.25) were more likely to be mentally unhealthy, whereas the female migrant workers who experienced more stress in "interpersonal tensions and conflicts" (OR 6.52, 95% CI 0.83-51.14) were more likely to have poor mental health. CONCLUSION: The findings provide information for the prevention of mental illness among migrant workers in China. The implications and limitations are also discussed. PMID- 18398560 TI - Portal hypertension and hypersplenism in a patient with a Bochdalek hernia: a case report. AB - BACKGROUND: A congenital diaphragmatic hernia (Bochdalek's hernia) rarely presents in adulthood. CASE PRESENTATION: We present the case of a 21-year-old woman with thrombocytopenia secondary to hypersplenism due to left-sided (sinistral) portal hypertension. This portal hypertension was caused by strangulation of the spleen within a Bochdalek's hernia. Subacute gastric volvulus compounded by portal hypertensive gastropathy resulted in her presentation with acute haematemesis. CONCLUSIONS: Congenital diaphragmatic hernias may present in the adult. Their presentation is variable, and they may produce life-threatening complications. PMID- 18398561 TI - Ancient DNA clarifies the evolutionary history of American Late Pleistocene equids. AB - Hippidions are past members of the equid lineage which appeared in the South American fossil record around 2.5 Ma but then became extinct during the great late Pleistocene megafaunal extinction. According to fossil records and numerous dental, cranial, and postcranial characters, Hippidion and Equus lineages were expected to cluster in two distinct phylogenetic groups that diverged at least 10 MY, long before the emergence of the first Equus. However, the first DNA sequence information retrieved from Hippidion fossils supported a striking different phylogeny, with hippidions nesting inside a paraphyletic group of Equus. This result indicated either that the currently accepted phylogenetic tree of equids was incorrect regarding the timing of the evolutionary split between Hippidion and Equus or that the taxonomic identification of the hippidion fossils used for DNA analysis needed to be reexamined (and attributed to another extinct South American member of the equid lineage). The most likely candidate for the latter explanation is Equus (Amerhippus) neogeus. Here, we show by retrieving new ancient mtDNA sequences that hippidions and Equus (Amerhippus) neogeus were members of two distinct lineages. Furthermore, using a rigorous phylogenetic approach, we demonstrate that while formerly the largest equid from Southern America, Equus (Amerhippus) was just a member of the species Equus caballus. This new data increases the known phenotypic plasticity of horses and consequently casts doubt on the taxonomic validity of the subgenus Equus (Amerhippus). PMID- 18398562 TI - An analysis of correspondence between unique rabies virus variants and divergent big brown bat (Eptesicus fuscus) mitochondrial DNA lineages. AB - The literature supports that unique rabies virus (RABV) variants are often compartmentalized in different species of bats. In Colorado, two divergent mtDNA lineages of big brown bats (Eptesicus fuscus) co-occur. RABV associated with this species also segregates into two clades. We hypothesized that unique RABV variants might be associated with mtDNA lineages of Colorado big brown bats. DNA was extracted from brain tissue of rabid big brown bats, the ND2 gene was amplified to determine mtDNA lineage, and the lineage was compared to a previously derived phylogenetic analysis of the RABV N gene. No correspondence was found between host bat lineage and RABV variant. PMID- 18398563 TI - Isolation, characterization and genome sequence of a birnavirus strain from flounder Paralichthys olivaceus in China. AB - A birnavirus strain, Paralichthys olivaceus birnavirus (POBV), was isolated and characterized from cultured flounder in China, and its complete genomic sequence was subsequently determined. The virus could induce cytopathic effects (CPE) in four of seven fish cell lines and was resistant to chloroform, 5-iodo-2' deoxyuridine, acid and alkaline pH, and heat treatment. Purified virus particles had a typical icosahedral shape, with a diameter of approximately 55-60 nm. The genomic segments A and B of POBV were 3,091 and 2,780 bp in length and shared many of the features of the members of the family Birnaviridae. Segment A contained two partially overlapping ORFs encoding a polyprotein, pVP2-VP4-VP3, and a nonstructural protein, VP5, while segment B had only one ORF encoding for the VP1, a viral RNA-dependent RNA polymerase (RdRp). This is the first report about a birnavirus strain from a new non-salmonid host in China and its complete genome sequence. PMID- 18398564 TI - A geometrical template for toroidal aggregates of chiral macromolecules. AB - The formation of toroidal aggregates by long chiral molecules of biological origin, as collagen, f-actin and DNA, or by chiral synthetic polypeptides has been observed in specific ionic environments. Such aggregates have received considerable attention in order to identify the various physical factors susceptible to contribute to this original morphogenesis, particularly in the case of those formed by DNA. We consider here the eventual role of a spontaneous uniform twist of micrometric pitch whose possible occurrence is suggested by some observations and by recent studies of DNA dense phases exhibiting cholesteric and "blue" phase structures. Following an approach inspired by the geometry and topology of fiber bundles, we show that the necessity to propagate such a twist in a dense bundle of fibers leads to the formation of aggregates having a toroidal shape and, in the case of the nanometric aggregates of DNA, characteristic sizes similar to those observed. PMID- 18398565 TI - New organic FET-like photoactive device, experiments and DFT modeling. AB - We present the possible construction of an organic FET-like photoactive device in which source-drain current through a phthalocyanine ( H(2)Pc film is affected by a photo-induced dipolar field in a photoactive "gate" electrode. The influence of the dipolar electric field on charge transfer between H(2)Pc molecules is modeled by DFT quantum-chemical calculations on H(2)Pc dimers and tetramers. PMID- 18398566 TI - Hydrodynamic regimes of active rotators at fluid interfaces. AB - We analyze the collective motion of suspensions of active rotators at low Reynolds numbers which interact hydrodynamically. We introduce a simple model for a rotator which allows us to classify the relevant dynamical regimes of the suspension. We characterize the collective velocity at which rotators displace and analyze its implications at long times, when these rotator suspensions diffuse. We analyze the differences with respect to diffusion in suspensions of passive particles, and assess the relevance of the Stokes-Einstein relation on rotators' diffusivity. PMID- 18398567 TI - Detachment of stretched viscoelastic fibrils. AB - New experimental results are presented about the final stage of failure of soft viscoelastic adhesives. A microscopic view of the detachment of the adhesive shows that after cavity growth and expansion, well adhered soft adhesives form a network of fibrils connected to expanded contacting feet which fail via a sliding mechanism, sensitive to interfacial shear stresses rather than by a fracture mechanism as sometimes suggested in earlier work. A mechanical model of this stretching and sliding failure phenomenon is presented which treats the fibril as a nonlinear elastic or viscoelastic rod and the foot as an elastic layer subject to a friction force proportional to the local displacement rate. The force on the stretched rod drives the sliding of the foot against the substrate. The main experimental parameter controlling the failure strain and stress during the sliding process is identified by the model as the normalized probe pull speed, which also depends on the magnitude of the friction and PSA modulus. In addition, the material properties, viscoelasticity and finite extensibility of the polymer chains, are shown to have an important effect on both the details of the sliding process and the ultimate failure strain and stress. PMID- 18398568 TI - Two-dimensional fluctuating vesicles in linear shear flow. AB - The stochastic motion of a two-dimensional vesicle in linear shear flow is studied at finite temperature. In the limit of small deformations from a circle, Langevin-type equations of motion are derived, which are highly nonlinear due to the constraint of constant perimeter length. These equations are solved in the low-temperature limit and using a mean-field approach, in which the length constraint is satisfied only on average. The constraint imposes non-trivial correlations between the lowest deformation modes at low temperature. We also simulate a vesicle in a hydrodynamic solvent by using the multi-particle collision dynamics technique, both in the quasi-circular regime and for larger deformations, and compare the stationary deformation correlation functions and the time autocorrelation functions with theoretical predictions. Good agreement between theory and simulations is obtained. PMID- 18398569 TI - Toxicity from chemoradiotherapy in older patients with glioblastoma multiforme. AB - INTRODUCTION: Elderly patients have glioblastomas (GBM) that are aggressive and poorly responsive to treatment. They are also prone to the side effects of treatment of GBM. METHODS: To shed light on the treatment of elderly patients with GBM, we reviewed the treatment toxicities and survival of patients 65 years of age or older who were treated with chemoradiotherapy, which is the new standard of care for GBM in younger patients. RESULTS: Thirty-nine patients at a single cancer center in Canada met the eligibility criteria for this retrospective study. Nineteen patients were treated initially with TMZ and radiotherapy and 20 others were treated with radiotherapy alone (only two had TMZ subsequently). Eight patients in the chemoradiotherapy group (42%) experienced Grade III or IV toxicity versus none in the radiotherapy group. The median overall survival in the chemoradiotherapy group was 8.5 months (range, 2.0-24.7 months) versus 5.2 months (range, 1.5-14.2 months) in the radiotherapy group, an apparent benefit which may have been due to an imbalance in age at diagnosis, extent of resection and performance status. In this series of GBM cases, methylation of the MGMT gene promoter was not associated with longer survival, either overall, or within the chemoradiotherapy treated subset. CONCLUSIONS: Elderly patients with GBM treated with chemoradiotherapy can be expected to experience significant toxicity. Large randomized trials will be necessary to determine whether chemoradiotherapy prolongs the survival of elderly patients and whether MGMT promoter status predicts benefit from temozolomide in this subset of patients. PMID- 18398570 TI - Intracranial Hodgkin's lymphoma in an HIV positive patient. AB - Intracranial Hodgkin's lymphoma (ICHL) in the HIV positive population is a rarely reported event with only three prior cases reported in the literature. In these prior reports, the outcome of the patients was uncertain or very poor. We report here the case of an HIV seropositive patient with an isolated intracranial recurrence of Hodgkin's Lymphoma (HL) who achieved long-term control of his cranial disease and continues to have a high performance status more than 3 years following treatment. PMID- 18398571 TI - Low grade astrocytoma presenting with visual loss. PMID- 18398572 TI - Cryptococcal meningitis in patients with glioma: a report of two cases. AB - OBJECTIVE AND IMPORTANCE: We describe two patients with high-grade glioma undergoing treatment with corticosteroids and chemotherapy who presented with cryptococcal meningitis and sepsis. This report of two cases highlights the importance of examining the efficacy of prophylactic antibiotic and/or antifungal regimens in this patient population due to their increased risk of opportunistic infections. CLINICAL PRESENTATION: A 73-year-old man with a history of glioblastoma multiforme (GBM), on dexamethasone and status post radiation therapy and two cycles of temozolamide, presented with decreased level of consciousness for 24 h and was found to have cerebrospinal fluid (CSF) and blood cultures positive for Cryptococcus neoformans. A 33-year-old man with a history of anaplastic astrocytoma, on dexamethasone and status post radiation therapy, four cycles of temozolomide and two cycles of Lomustine (CCNU), presented with headache, dizziness and photophobia and was found to have CSF and blood cultures positive for Cryptococcus neoformans. INTERVENTION: Both patients were treated with an initial regimen of amphotericin B and flucytosine for a minimum of two weeks and switched to fluconazole for 6 months to 1 year of treatment. CONCLUSION: Patients with high-grade glioma treated with long-term corticosteroid therapy and chemotherapy are at increased risk of developing opportunistic infections. The two patients reported here developed cryptococcal meningitis and sepsis. Prophylactic regimens with either fluconazole or itraconazole currently exist that effectively decrease the incidence of both cryptococcal infections. Further investigations into the risk:benefit ratio of primary prophylactic therapy in this patient population may prove beneficial. PMID- 18398573 TI - Gene expression profile analysis of primary glioblastomas and non-neoplastic brain tissue: identification of potential target genes by oligonucleotide microarray and real-time quantitative PCR. AB - The prognosis of glioblastomas is still extremely poor and the discovery of novel molecular therapeutic targets can be important to optimize treatment strategies. Gene expression analyses comparing normal and neoplastic tissues have been used to identify genes associated with tumorigenesis and potential therapeutic targets. We have used this approach to identify differentially expressed genes between primary glioblastomas and non-neoplastic brain tissues. We selected 20 overexpressed genes related to cell cycle, cellular movement and growth, proliferation and cell-to-cell signaling and analyzed their expression levels by real time quantitative PCR in cDNA obtained from microdissected fresh tumor tissue from 20 patients with primary glioblastomas and from 10 samples of non neoplastic white matter tissue. The gene expression levels were significantly higher in glioblastomas than in non-neoplastic white matter in 18 out of 20 genes analyzed: P < 0.00001 for CDKN2C, CKS2, EEF1A1, EMP3, PDPN, BNIP2, CA12, CD34, CDC42EP4, PPIE, SNAI2, GDF15 and MMP23b; and NFIA (P: 0.0001), GPS1 (P: 0.0003), LAMA1 (P: 0.002), STIM1 (P: 0.006), and TASP1 (P: 0.01). Five of these genes are located in contiguous loci at 1p31-36 and 2 at 17q24-25 and 8 of them encode surface membrane proteins. PDPN and CD34 protein expression were evaluated by immunohistochemistry and they showed concordance with the PCR results. The present results indicate the presence of 18 overexpressed genes in human primary glioblastomas that may play a significant role in the pathogenesis of these tumors and that deserve further functional investigation as attractive candidates for new therapeutic targets. PMID- 18398574 TI - Brain metastases in breast cancer: clinical and pathologic characteristics associated with improvements in survival. AB - BACKGROUND: As breast cancer patients live longer with control of systemic disease, survival after the diagnosis of brain metastases (BM) also appears to be improving. METHODS: The authors conducted a retrospective review of 112 breast cancer patients diagnosed with BM from 1997 to 2007 and correlated clinical and pathologic characteristics including hormone receptor (HR) and Her2/neu status with outcomes. FINDINGS: Median time to BM diagnosis (TTBM) was 38 months (range, 0-204 months). TTBM was shorter for patients with HR- versus HR+ disease (median 28.8 vs. 61.2 months, P < 0.001, Wilcoxon test). No difference in TTBM was observed for patients with HER2- versus HER2+ disease (median 37.4 vs. 34.9 months, P = 0.81). Median survival after the diagnosis of BM was 14.4 months. There was no significant difference in median survival after BM diagnosis for patients with HR+ versus HR- cancers (19.9 vs. 11.0 months, P = 0.18, log rank) or for patients with HER2+ versus HER2- disease (23.1 vs. 13.3 months, P = 0.11, log rank). Survival was significantly longer in patients with stable or responding systemic disease at BM diagnosis compared to patients with progressing systemic disease (31 vs. 6.3 months, P < 0.001). Multivariate analysis revealed that HR positivity, age <50, Karnofsky Performance Score (KPS) > or =80, and stable or responding systemic disease at BM diagnosis were associated with improved survival. INTERPRETATION: Subsets of patients with breast cancer BM are surviving longer. Control of systemic disease was most strongly associated with improved outcomes, and HER2/neu overexpression did not shorten survival after the diagnosis of BM. PMID- 18398575 TI - Selective cancer-germline gene expression in pediatric brain tumors. AB - Cancer-germline genes (CGGs) code for immunogenic antigens that are present in various human tumors and can be targeted by immunotherapy. Their expression has been studied in a wide range of human tumors in adults. We measured the expression of 12 CGGs in pediatric brain tumors, to identify targets for therapeutic cancer vaccines. Real Time PCR was used to quantify the expression of genes MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A10, MAGE-A12, MAGE-C2, NY-ESO-1 and GAGE-1,2,8 in 50 pediatric brain tumors of different histological subtypes. Protein expression was examined with immunohistochemistry. Fifty-five percent of the medulloblastomas (n = 11), 86% of the ependymomas (n = 7), 40% of the choroid plexus tumors (n = 5) and 67% of astrocytic tumors (n = 27) expressed one or more CGGs. Immunohistochemical analysis confirmed qPCR results. With exception of a minority of tumors, the overall level of CGG expression in pediatric brain tumors was low. We observed a high expression of at least one CGG in 32% of the samples. CGG-encoded antigens are therefore suitable targets in a very selected group of pediatric patients with a brain tumor. Interestingly, glioblastomas from adult patients expressed CGGs more often and at significantly higher levels compared to pediatric glioblastomas. This observation is in line with the notion that pediatric and adult glioblastomas develop along different genetic pathways. PMID- 18398577 TI - Beyond platelet inhibition: potential pleiotropic effects of ADP-receptor antagonists. AB - The adenosine diphosphate (ADP) receptor antagonists, specifically the class of thienopyridines, have emerged as potent tools in the clinician's arsenal for the treatment of athero-thrombotic disease over the last two decades. Though these medications have been clearly demonstrated to have significant platelet inhibiting effects, their potential positive impact on other systemic processes has been less well elucidated. Recent evidence points to a number of potential pleiotropic effects of these agents, most notably in the attenuation of several pro-inflammatory pathways, which may be independent of their anti-platelet aggregating effect. Additionally, several new ADP receptor antagonists are under investigation; it remains to be seen if these agents possess any additional beneficial pleiotropic properties as well. PMID- 18398578 TI - Advanced proliferative diabetic retinopathy in four young patients with flexion contractures (Rosenbloom syndrome). AB - In this report, we present four patients with type 1 diabetes of long duration (more than 20 years) who had multiple joint contractures (Rosenbloom syndrome) and who presented with reduced vision and advanced proliferative diabetic retinopathy. All of the cases had short stature which was non-familial and all had other microvascular complications (nephropathy and neuropathy). It seems that limited joint mobility (LJM) is a risk factor not only for microvascular complications, but also for more severe and advanced proliferative retinopathy. PMID- 18398576 TI - BEHAB/brevican requires ADAMTS-mediated proteolytic cleavage to promote glioma invasion. AB - Malignant gliomas are the most common and deadly primary brain tumors, due to their infiltrative invasion of the normal neural tissue that makes them virtually impossible to completely eliminate. We have previously identified and characterized the proteoglycan BEHAB/brevican in gliomas and have demonstrated that upregulation and cleavage of this CNS-specific molecule promote glioma invasion. Here, we have further investigated if the proteolytic processing of BEHAB/brevican by metalloproteases of the ADAMTS family is a necessary step in mediating its pro-invasive effect in glioma. By generating a site-specific ((396)SRG(398) --> NVY) mutant form resistant to ADAMTS cleavage, we have shown that the predominant proteolytic processing of BEHAB/brevican by glioma cells occurs only at this site. More importantly, "uncleavable" BEHAB/brevican is unable to enhance glioma cell invasion in vitro and tumor progression in vivo. In addition, our results suggest that the full-length protein and its cleavage products may act independently because the mutant form does not exert a dominant negative effect on normal BEHAB/brevican expression or cleavage. These results illustrate how the regulated processing of major components of the neural extracellular matrix has important functional implications in glioma progression. In addition, our findings underscore the relevance of the ADAMTS family of metalloproteases as attractive targets for novel pharmacological approaches in glioma therapy. PMID- 18398579 TI - Atrophy of the retinal pigment epithelium following vitrectomy with trypan blue. AB - PURPOSE: To report a case of possible trypan blue retinotoxicity after macular hole surgery. METHODS: Retrospective review of case notes and follow-up in one patient with unusual appearance of the fundus after macular hole surgery with trypan blue. RESULTS: A 69-year-old female presented to our eye clinic with a complaint of loss of vision over the last 6 months. Examination showed a full thickness macular hole in the right eye. She underwent right pars plana vitrectomy with internal limiting peel assisted by trypan blue. Surgery was routine with no difficulty during surgical peel of the inner limiting membrane. Two weeks postoperatively, increased pigmentation and atrophy at the level of the retinal pigment epithelium (RPE) was noted temporal to the fovea. The macular hole appeared closed. Fundus fluorescein angiography showed this to be a lesion with characteristics of RPE atrophy. CONCLUSION: In the absence of excessive peroperative surgical trauma to the region of the scar, the retinal changes may be secondary to possible toxicity due to trypan blue stain, light toxicity or a combination of both. PMID- 18398580 TI - Stone silent. PMID- 18398581 TI - Systemic AL amyloidosis with disseminated intravascular coagulation associated with hyperfibrinolysis. AB - Many coagulation abnormalities are known to coexist in patients with AL amyloidosis; however, disseminated intravascular coagulation (DIC) is rarely observed. We describe the case of a 61-year-old woman who presented with systemic purpura, macroscopic hematuria, and hepatosplenomegaly as the initial manifestations of systemic AL amyloidosis. A coagulation study revealed severe DIC associated with fibrinolysis. The patient was treated for DIC with gabexate mesilate (GM); however, her bleeding symptoms and thrombocytopenia continued to worsen. The treatment was changed from GM to nafamostat mesilate (NM); DIC improved gradually, and the platelet count normalized in 1 week. After the tapering and cessation of NM therapy, deterioration of DIC did not occur. She underwent autologous peripheral blood stem cell transplantation twice following high-dose melphalan therapy, and received maintenance therapy with thalidomide. Hepatosplenomegaly progression appears to have been halted, and DIC has not recurred. This is the first reported case of AL amyloidosis showing severe DIC with excessive fibrinolysis. The clinical observation that NM was considerably more effective than GM in our patient suggests that NM may be more suitable for the treatment of DIC with a hyperfibrinolytic condition in AL amyloidosis patients. PMID- 18398582 TI - Comparison of 60cobalt and 192iridium sources in high dose rate afterloading brachytherapy. AB - PURPOSE: (60)Co sources with dimensions identical to those of (192)Ir have recently been made available in clinical brachytherapy. A longer half time reduces demands on logistics and quality assurance and perhaps costs. MATERIAL AND METHODS: Comparison of the physical properties of (60)Co and (192)Ir with regard to brachytherapy. RESULTS: Required activities for the same air kerma rate are lower by a factor of 2.8 for (60)Co. Differential absorption in tissues of different densities can be neglected. Monte Carlo calculations demonstrate that integral dose due to radial dose fall off is higher for (192)Ir in comparison to (60)Co within the first 22 cm from the source (normalization at 1 cm). At larger distances this relationship is reversed. CONCLUSION: Clinical examples for intracavitary and interstitial applications however, show practically identical dose distributions in the treatment volume. PMID- 18398583 TI - Whole lung irradiation in patients with exclusively pulmonary metastases of Ewing tumors. Toxicity analysis and treatment results of the EICESS-92 trial. AB - BACKGROUND: In the European Intergroup Cooperative Ewing's Sarcoma Study (EICESS) 92, whole lung irradiation (WLI) was performed in patients with primary lung metastases. This retrospective analysis evaluates the pulmonary function and the outcome of patients with exclusively pulmonary metastases. PATIENTS AND METHODS: Between 1990 and 1999, 99 patients were registered into the EICESS-92-study trial with exclusively pulmonary metastases of Ewing tumors. The multimodal treatment regimen included polychemotherapy and local therapy to the primary tumor. WLI was performed with a dose between 12-21 Gy. 70 patients were treated with WLI, 13 of them received a further boost to their primary tumor in the thorax up to a cumulative dose of 54 Gy. RESULTS: Pulmonary function tests were available for 37 patients treated with WLI (+/- boost). None, mild, moderate or severe pulmonary complications were seen in 43%, 29%, 21% and 7% of patients treated with WLI without further boost (median follow-up 25.2 months). Patients with an additional radiation boost or surgery to the thorax showed slightly higher rates of complications. Overall survival (OAS) showed a trend towards better results for patients with WLI (5-year-OAS: 0.61 for WLI vs. 0.49 for no WLI, p = 0.36). CONCLUSION: These data indicate a benefit and acceptable toxicity for WLI in the presented collective of patients. As long as there is no randomized prospective analysis, the present data confirm the indication for WLI in Ewing tumor patients with primary exclusively lung metastases. PMID- 18398584 TI - Evaluation of prognostic factors and two radiation techniques in patients treated with surgery followed by radio(chemo)therapy or definitive radio(chemo)therapy for locally advanced head-and-neck cancer. AB - BACKGROUND AND PURPOSE: Conventional radiotherapy (RT) still is the standard technique for head-and-neck cancer in many centers worldwide, whereas other centers replaced this technique by 3-D conformal RT, which is associated with more appropriate dose distributions. Comparative studies regarding outcome and toxicity are lacking. This study compared both techniques for overall survival (OS), metastases-free survival (MFS), loco-regional control (LC), and toxicity in stage III/IV head-and-neck cancer. PATIENTS AND METHODS: Data of 345 patients irradiated for stage III/IV squamous cell head-and-neck cancer were retrospectively analyzed. Patients received conventional RT (group A, n = 166) or 3-D conformal RT (group B, n = 179). Both techniques were compared for outcomes and toxicity. Eleven further potential prognostic factors were investigated: age, gender, performance status, tumor site, grading, T-stage, N-stage, AJCC-stage, chemotherapy, surgery, pre-RT hemoglobin. RESULTS: 3-year-OS was 62% in group A and 57% in group B (p = 0.15). 3-year-MFS was 67% and 76% (p = 0.46), 3-year-LC was 65% and 68%, respectively (p = 0.71). On multivariate analysis, gender (p = 0.005), performance status (p < 0.001), T-stage (p = 0.002), and N-stage (p < 0.001) were associated with OS. MFS was influenced by performance status (p < 0.001) and N-stage (p < 0.001), LC by gender (p = 0.021), T-stage (p < 0.001), and pre-RT hemoglobin level (>or= 12 better than < 12 g/dl, p = 0.004). Grade 2-3 xerostomia was less frequent with 3-D conformal RT (43% vs. 58%, p = 0.06). Otherwise, toxicities were similar. CONCLUSION: Both RT techniques resulted in similar treatment outcomes. Because xerostomia was less with 3-D conformal RT, this technique appeared beneficial for patients, in whom one parotid gland can be spared. Outcome was associated with gender, performance status, tumor stage, and pre-RT hemoglobin. PMID- 18398585 TI - Lymph edema of the lower extremities after lymphadenectomy and radiotherapy for cervical cancer. AB - PURPOSE: To assess the incidence of clinical lymph edema after lymphadenectomy and postoperative radiotherapy (RT). PATIENTS AND METHODS: From 1994-2002 192 patients with risk factors for recurrence received radiotherapy with FIGO I (58.8%), II (35.4%), III (4.2%) or IV (1.6%). RT consisted of teletherapy (10.4%), brachytherapy of the vaginal vault (20.8%) or a combination of both (68.8%). Additional chemotherapy was given in 69 patients (35.9%). Surgery comprised laparoscopically assisted radical vaginal hysterectomy (LARVH) (35.4%), radical abdominal hysterectomy (RAH) (48.4%), simple hysterectomy (HE) (11.5%) or exenteration (4.7%). RESULTS: 73 patients had lymph node metastases, 119 had negative lymph nodes. In patients with LARVH 6-74 (median 30) lymph nodes were removed, with RAH 3-70 (median 21 lymph nodes), and after HE or exenteration 5-50 (median 13 lymph nodes). 90 patients had 25 or less lymph nodes removed, 83 patients more than 25 lymph nodes removed. Prognostic factors, such as age, FIGO stages, histologic grading and type of histology were well balanced in these cohorts. 45 (23.4%) of all patients developed clinically relevant lymph edema of the lower limb with a median latency of 11 (1-121) months. When 25 or less lymph nodes were removed 17.8% of patients developed leg edema, when more than 25 lymph nodes were removed 32.5% of patients were diagnosed with lymph edema (p = 0.025). Radiotherapy and chemotherapy had no influence on the incidence of leg edema. Overall survival at 5 (10) years was independent of number of lymph nodes removed. CONCLUSION: The data suggest increasing rates of leg edema with increasing number of lymph nodes dissected independent of the type of radiotherapy and chemotherapy performed. The lymph node sampling policy should be planned carefully in respect to minimize the risk of leg lymph edema. PMID- 18398586 TI - Prophylaxis of heterotopic ossification in patients sedated after polytrauma : medical and ethical considerations. AB - BACKGROUND AND PURPOSE: Heterotopic ossification (HO) often follows acetabular fractures after multitrauma. Irradiation is a mean for prophylaxis. We established a standard procedure in our hospital for patients under sedation, when obtaining informed consent for HO prophylaxis is impossible. PATIENTS AND METHODS: We reviewed current scientific evidence, calculated the risks of radiation and presented the ethical and legal framework. The subject was scrutinised by an interdisciplinary panel. RESULTS: Irradiation is the most effective means for prophylaxis and has few adverse effects in adult patients with fractures of the acetabulum. The lifetime risk of radiation-induced cancer or infertility are insignificant. CONCLUSIONS: Informed consent for irradiation should be obtained before operation whenever possible. When this cannot be done prophylaxis can be postponed for a maximum of 3 days in order to obtain consent. If the patient is not able to communicate within this period, prophylactic irradiation should be given after consulting the relatives. The patient must be informed as soon as possible. PMID- 18398587 TI - Long-term results of radiotherapy in patients with chronic palmo-plantar eczema or psoriasis. AB - BACKGROUND AND PURPOSE: Radiotherapy (RT) is well accepted for therapy-refractory palmo-plantar eczema or psoriasis, despite of lacking evidence regarding beneficial long term effects. Furthermore, the optimal irradiation dose is unknown. We evaluated the outcome of RT with two different RT single/total dose (SD/TD) treatment policies. PATIENTS AND METHODS: 28 consecutive patients with therapy-refractory eczema (n = 22) or psoriasis (n = 6) of palms and/or soles were irradiated twice a week either with a D(max) SD of 1 Gy (6/98-5/03; median TD: 12 Gy) or 0.5 Gy (6/03-7/04; median TD: 5 Gy). Median age was 52 years (27 71), median follow-up 20 months (4-76). Totally 88 regions were treated, 49 with 1 Gy, 39 with 0.5 Gy SD. Eight different symptoms were scored from 0 (absent) -3 (severe), giving a possible sum score of 0-24. Patients' rating of RT result was also documented (worse/stable/better/complete remission). RESULTS: The sum score was 15 (6-23) before RT, 2 (0-16) at the end of RT, and 1 (0-21) at last follow up, respectively. The improvement was highly significant in both treatment regimens. Better or complete remission by the patients were reported in 44 and 39 (= 83 out of 88) localisations, that was often stable during the follow-up. 5 (6%) regions in 3 (11%) patients didn't benefit from RT. CONCLUSION: RT reveals excellent results in palmo-plantar eczema or psoriasis. We recommend a SD of 0.5 Gy twice a week up to a TD of 4-5 Gy. PMID- 18398588 TI - Painful spinal cord compression as a complication of extramedullary hematopoiesis associated with beta-thalassemia intermedia. AB - BACKGROUND: Spinal cord compression due to extramedullary hematopoiesis (EMH) is a rare manifestation of thalassemia. General therapeutic recommendations do not exist. CASE REPORT: We report a 23 year old male with beta-thalassemia intermedia and back pain resistant to analgetic medication. Multiple extradural mass lesions on MRI of the lumbar spine were identified. Hypertransfusion was initiated with no relief of symptoms. Low dose irradiation with 20 Gy had to be added to achieve adaequate response. RESULT: With the combination of transfusion therapy and irradiation a rapid and long-lasting relief of symptoms was achieved. 12 month post irradiation the patient is free of pain and neurological symptoms. CONCLUSION: Clinical awareness of this phenomenon and knowledge of the available treatment option is essential for successful outcome. In treatment refractory symptoms irradiation could successfully contribute to a rapid therapeutic response. PMID- 18398589 TI - Characteristics of the respiratory mechanical and muscle function of competitive breath-hold divers. AB - Competitive breath-hold divers (BHD) employ glossopharyngeal insufflation (GI) to increase intrapulmonary oxygen stores and prevent the lungs from dangerous compressions at great depths. Glossopharyngeal insufflation is associated with inflation of the lungs beyond total lung capacity (TLC). It is currently unknown whether GI transiently over-distends the lungs or adversely affects lung elastic properties in the long-term. Resting lung function, ventilatory drive, muscle strength, and lung compliance were measured in eight BHD who performed GI since 5.5 (range 2-6) years on average, eight scuba divers, and eight control subjects. In five BHD subsequent measures of static lung compliance (Cstat) were obtained after 1 and 3 min following GI. Breath-hold divers had higher than predicted ventilatory flows and volumes and did not differ from control groups with regard to gas transfer, inspiratory muscle strength, and lung compliance. A blunted response to CO2 was obtained in BHD as compared to control groups. Upon GI there was an increase in mean vital capacity (VCGI) by 1.75 +/- 0.85 (SD) L compared to baseline (p < 0.001). In five BHD Cstat raised from 3.7 (range 2.9-6.8) L/kPa at baseline to 8.1 (range 3.4-21.2) L/kPa after maximal GI and thereafter gradually decreased to 5.6 (range 3.3-8.1) L/kPa after 1 min and 4.2 (range 2.7-6.6) L/kPa after 3 min (p < 0.01). We conclude that in experienced BHD there is a transient alteration in lung elastic recoil. Resting lung function did not reveal a pattern indicative of altered lung ventilatory or muscle function. PMID- 18398591 TI - [The history of lymphoma classifications with special consideration of cutaneous lymphomas]. AB - For modern evidence-based medicine, classification systems are necessary to guarantee a uniform approach for therapy and for estimating prognosis. The comparability of clinical studies and international communication require a common language and are only possible with modern international classification systems. But because all classifications are artificial, they only mirror the current state of knowledge and may change dramatically over decades. This review discusses the history of lymphoma classifications systems with a special focus on the topic of primary cutaneous lymphomas, emphasizing special problems in terminology. PMID- 18398592 TI - [Surrogate endpoint trials: benefit and pitfalls for clinical decision making]. AB - Ideally clinicians should base their treatment decisions on results from randomised controlled trials which include patient-important outcomes, such as quality of life, prevented disease events or death. Conducting such trials often involves large sample sizes and extended follow-up periods. Therefore, researchers have aimed to conduct trials with surrogate endpoints by substituting patient-important outcomes in order to reduce sample size and observation time. Surrogate endpoints are outcomes that substitute for direct measures of how a patient feels, functions, or survives. In many countries drugs are approved based on data from surrogate endpoint trials. Recently, a controversy evolved on the reliability of results generated from these trials driven by unanticipated side effects or severe toxicity leading to the withdrawal of drugs that were solely approved based on evidence from surrogate endpoint trials. We present some recent examples and criteria how clinicians can critically evaluate the validity of claims by experts or the pharmaceutical industry in regard to the expected patients' benefit from drugs approved by results from surrogate endpoint trials. PMID- 18398593 TI - AT1-receptor autoantibodies and uteroplacental RAS in pregnancy and pre eclampsia. AB - Pre-eclampsia is a common, pregnancy-induced disorder, consisting of hypertension and proteinuria. The condition is one of the leading causes for maternal and perinatal morbidity and mortality. Nonetheless, the underlying molecular mechanisms remain unclear. Immunological mechanisms and the renin-angiotensin system have been implicated in the development of pre-eclampsia. Agonistic autoantibodies to the angiotensin II type I receptor (AT1-AA) have been identified in pre-eclamptic patients, unifying the two hypothesis. Evidence has also accumulated for the existence and importance of a local, utroplacental renin angiotensin system. We summarize recent data emphasizing the pathophysiological role for the autoantibodies and the uteroplacental renin-angiotensin system. PMID- 18398594 TI - [Diagnostic radiology of the pelvis. Prostate cancer, bladder cancer, and incontinence]. AB - Prostate cancer, bladder cancer, and pelvic floor weakness are among the most common diseases of the pelvis. Cardinal symptoms include painless macrohematuria in bladder cancer and urinary and fecal incontinence in pelvic floor weakness. Suspicion of prostate cancer currently is most frequently raised when the serum concentration of prostate-specific antigen is pathologically elevated. Besides extensive clinical and invasive diagnosis, clinical imaging is frequently applied for the localization, locoregional staging, and diagnosis of recurrence of prostate cancer and invasive bladder cancer, and in clinically difficult cases of cystocele, enterocele, rectocele, descensus or prolapse of vagina, uterus, and rectum, and rectal intussusception. Magnetic resonance imaging with T2-weighted TSE or FSE images in several planes combined with either axial, T1-weighted images and MR spectroscopy for the prostate, dynamic contrast-enhanced T1 weighted images for the urinary bladder, or dynamic T2-weighted functional images for pelvic floor incontinence are particularly well suited as clinical imaging methods. PMID- 18398595 TI - [Epidemiology of prostate cancer: recent results from the Epidemiological Cancer Register of the District of Munster (Germany)]. AB - BACKGROUND: Prostate cancer is characterized by worldwide increasing incidence rates, improved survival, and decreasing mortality. We investigated the current situation in the Epidemiological Cancer Register of the District of Munster, Germany (which has approximately 1.25 million male inhabitants). MATERIALS AND METHODS: We calculated the incidence and mortality rates, stage distribution, and relative survival rates for prostate cancer between the years 2002 and 2004. RESULTS: The age-standardized incidence rate was 115/100,000 men per year, and the median age at diagnosis was 70 years. The tumour stage was T1/T2 in 69.6% of cases. The estimated relative survival after 5 years was 83.5% (95% confidence interval 81.4-85.4) and after 10 years was 73.3% (69.5-77.0). Survival was barely affected when the tumour was limited to the prostate (UICC I-II), whereas survival rates were markedly reduced when the tumour had spread or had infiltrated adjacent structures (UICC IV; relative 10-year survival rate 22.1%). CONCLUSIONS: The majority of patients with prostate cancer now have a favourable prognosis. Increased incidence rates must be interpreted in the context of widespread use of prostate-specific antigen testing. PMID- 18398596 TI - [Bladder carcinoma cell lines as models of the pathobiology of bladder cancer. Review of the literature and establishment of a new progression series]. AB - BACKGROUND: Tumour cell lines represent valuable preclinical models to decipher underlying biology and identify potential therapy targets and pharmacologically useful compounds. Approximately 50 human bladder cancer cell lines have been established to date, mainly from invasive and metastatic tumours. Two of these, namely T24 and 253J, were experimentally further developed into progression series. These models have provided important insights into later tumour progression events and metastatic dissemination. Only a few cell lines are available as models of non-invasive papillary bladder cancer and no progression series have yet been established. MATERIAL AND METHODS: During the course of establishing a doxorubicin-resistant variant cell line of the human papillary bladder carcinoma cell line BFTC-905, a unique cell colony was identified, apparently involving cells with divergent growth patterns. Subsequent subculturing yielded three daughter cell lines, BFTC-905-compact, BFTC-905 diffuse und BFTC-905-diffuse M. Their fundamental characterization included basic cell morphology, cell membrane expression of E-Cadherin, karyotype analysis, invasion and colony forming capacity in soft agar. The clonal origin of the newly established daughter cell lines was assessed by means of molecular genetic methods. RESULTS: We could identify important differences in multiple transformation related traits among the cell lines of the BFTC-905 progression series. Both diffuse cell lines (BFTC-905-diffuse und BFTC-905-diffuse M) differed from the BFTC-905-compact cell line by growing in a less organized,"diffuse" manner, which involved colonies of cells exhibiting apparently normal cell-to-cell adhesion as well as individual cells outside of them. This diminution of the cell-to-cell adhesion was accompanied by a corresponding decrease of membranous E-Cadherin. The BFTC-905-diffuse M cell line displayed a dramatic increase in the overall chromosome number, resulting in a hypertetraploid karyotype. At the same time, this cell line, as the only one in the progression series, acquired the ability to grow independent of anchorage in soft agar. All three cell lines remained noninvasive. Allelic distribution of highly polymorphic DNA-markers in the cell lines of the BFTC-905 progression series provided unequivocal evidence of their common origin. CONCLUSION: The newly established BFTC-905 progression series manifests two aspects of the early progression of non-invasive bladder carcinoma, not exhibited by any other progression series published so far, namely dynamic changes in the expression of E-Cadherin and a complex karyotypic evolution. It may thus contribute important insights into further understanding of the pathobiology of bladder cancer. PMID- 18398597 TI - [Ossification in chronic Achilles tendinosis: a third calf bone]. AB - Ossification of the Achilles tendon is a rare complication of chronic Achilles tendinosis. This case report discusses a 59-year-old man who experienced Achilles tendon lengthening in early childhood due to a congenital clubfoot disorder with a record-breaking ossification of a length of 10 cm. Because of persistent symptoms and the risk of fracture, the choice of treatment after unsuccessful conservative therapy depends on surgical reconstructive criteria. PMID- 18398598 TI - NICEM consensus on neurological monitoring in acute neurological disease. AB - This manuscript summarises the consensus on neuromonitoring in neuro-intensive care promoted and organised by the Neuro-Intensive Care and Emergency Medicine (NICEM) Section of the European Society of Intensive Care Medicine (ESICM). It is expected that continuous monitoring using multi-modal techniques will help to overcome the limitations of each individual method and will provide a better diagnosis. More specific treatment can then be applied; however, it remains to be determined which combination of parameters is optimal. The questions discussed and addressed in this manuscript are: (1) Who should have ICP monitoring and for how long? (2) What ICP technologies are available and what are their relative advantages/disadvantages? (3) Should CPP monitoring and autoregulation testing be used? (4) When should brain tissue oxygen tension (PbrO(2)) be monitored? (5) Should structurally normal or abnormal tissue be monitored with PbrO(2)? (6) Should microdialysis be considered in complex cases? It is hoped that this document will prove useful to clinicians working in NICU and also to those developing specialist NICU services within their hospital practice. PMID- 18398599 TI - Dipeptidyl peptidase IV inhibitors in diabetes: more than inhibition of glucagon like peptide-1 metabolism? AB - Inhibitors of the protease dipeptidyl peptidase IV (DPP-IV) are promising new drugs for the treatment of type 2 diabetes. They are thought to act by inhibiting the breakdown of glucagon-like peptide-1 and, thereby, selectively enhancing insulin release under conditions when it is physiologically required. These drugs are selective for DPP-IV, but the enzyme itself has a broad range of substrates other than glucagon-like peptide-1. Other high affinity substrates of DPP-IV including peptide YY may also play a role in the regulation of energy homeostasis. Moreover, DPP-IV is also known as CD26 and considered to be a moonlighting protein because it has a wide range of other functions unrelated to energy homeostasis, e.g. in immunity. The potential role of DPP-IV inhibition on substrates other than glucagon-like peptide-1 in diabetes patients remains to be elucidated. PMID- 18398600 TI - Pharmacological profile of ASP8497, a novel, selective, and competitive dipeptidyl peptidase-IV inhibitor, in vitro and in vivo. AB - Dipeptidyl peptidase (DPP)-IV is involved in the inactivation of glucagon-like peptide-1 (GLP-1), a potent insulinotropic peptide. Thus, DPP-IV inhibitors are expected to become a useful new class of antidiabetic agent. This report describes the pharmacological profile of the novel DPP-IV inhibitor, ASP8497 [(2S,4S)-4-fluoro-1-({[4-methyl-1-(methylsulfonyl)piperidin-4 yl]amino}acetyl)pyrrolidine-2-carbonitrile monofumarate], both in vitro and in vivo. ASP8497 inhibited DPP-IV in plasma from mice, dogs, and humans with median inhibition concentration (IC(50)) values of 2.6 nM, 7.3 nM, and 6.2 nM, respectively. In contrast, ASP8497 did not potently inhibit human DPP8 or DPP9 activity (IC(50)=1,700 nM and 100 nM, respectively) and exhibited selectivity against several proteases, including proline-specific proteases (IC(50)>10 microM). Kinetic analysis indicated that ASP8497 is a competitive DPP-IV inhibitor. In normal mice, ASP8497 inhibited plasma DPP-IV activity even 12 h after administration. ASP8497 significantly inhibited increases in the blood glucose level during the oral glucose tolerance test (OGTT) conducted 0.5 h after administration. This was accompanied by increases in the plasma active GLP-1 and insulin levels. In addition, ASP8497 significantly inhibited increases in the blood glucose level during the OGTT conducted 8 h after administration. Furthermore, in Zucker fatty rats, ASP8497 dose dependently improved glucose tolerance with significance at doses of 1 mg/kg or higher. In contrast, ASP8497 did not cause hypoglycemia in fasted normal mice. These results indicate that ASP8497 is a potent, competitive, and selective DPP-IV inhibitor with antihyperglycemic activity. PMID- 18398602 TI - Reference measurement procedure for Delta9-tetrahydrocannabinol in serum. AB - An isotope dilution gas chromatography/mass spectrometry (ID-GC/MS) reference measurement procedure for Delta9-tetrahydrocannabinol (THC) in serum was developed and validated. The method complies with the concept of a ratio primary reference measurement procedure. The uncertainty was determined for two concentrations of THC in serum (1 ng/mL and 2.4 ng/mL). The calculation procedure is based on the Guide to the Expression of Uncertainty in Measurement (GUM). The relative expanded uncertainty was found to be less than 2% for both concentration levels, corresponding to a 95% confidence interval. For the reference method, it was shown that the measurement of THC within the concentration range covered by the current threshold values is very accurate. The method has the potential to provide traceability for the methods used in practical forensics. PMID- 18398603 TI - Catechol determination in compost bioremediation using a laccase sensor and artificial neural networks. AB - An electrochemical biosensor based on the immobilization of laccase on magnetic core-shell (Fe(3)O(4)-SiO(2)) nanoparticles was combined with artificial neural networks (ANNs) for the determination of catechol concentration in compost bioremediation of municipal solid waste. The immobilization matrix provided a good microenvironment for retaining laccase bioactivity, and the combination with ANNs offered a good chemometric tool for data analysis in respect to the dynamic, nonlinear, and uncertain characteristics of the complex composting system. Catechol concentrations in compost samples were determined by using both the laccase sensor and HPLC for calibration. The detection range varied from 7.5 * 10(-7) to 4.4 * 10(-4) M, and the amperometric response current reached 95% of the steady-state current within about 70 s. The performance of the ANN model was compared with the linear regression model in respect to simulation accuracy, adaptability to uncertainty, etc. All the results showed that the combination of amperometric enzyme sensor and artificial neural networks was a rapid, sensitive, and robust method in the quantitative study of the composting system. PMID- 18398604 TI - Antioxidative effect of Bacteroides thetaiotaomicron extracts: superoxide dismutase identification. AB - Bacteroides thetaiotaomicron, a bowel anaerobic commensal, seems to release enzymes detoxifying reactive oxygen species according to our recent work. This opportunistic pathogen would be beneficial in the case of an inflammatory process. To explore its role after an oxidative or nutritive stress, six to seven separate experiments were performed. The bacteria were grown on media restricted in growth factors or supplemented with bile. Their viability was checked after surface protein extraction. The extracts underwent 2D electrophoresis. Gel images were statistically analysed to construct "master" gels. Proteins were identified (peptide-mass fingerprinting technique). The effect of each extract on superoxide anions was evaluated (spectrophotometric method). Superoxide dismutase was identified and a major superoxide anion inhibition was shown by extracts obtained after a nutritive and oxidative stress without significant bacterial death. So, a therapeutic antioxidant potential is firmly hoped for. [figure: see text] PMID- 18398605 TI - Characterisation of historical organic dyestuffs by liquid chromatography-mass spectrometry. AB - This review discusses the characterisation of natural organic dyestuffs of historical interest by liquid chromatography-mass spectrometry. The structures of the most important natural organic dyestuffs traditionally used are presented and discussed from the perspective of their analytical chemical determination. The practical aspects of the determination of this inhomogeneous range of compounds with different structures, such as anthraquinones, flavonoids, indigoids or tannins, are discussed with their implications for sample preparation, liquid chromatographic separation and mass spectrometric detection. The particular focus of this review is the discussion of the mass spectral fragmentation patterns of the different classes of natural organic dyestuffs, which in the ideal case allow the identification of the dyestuff actually used, and thereby provide a key to the better characterisation and understanding of historical objects dyed with natural organic dyestuffs. PMID- 18398606 TI - Can we produce an image of bacteria with radiopharmaceuticals? PMID- 18398601 TI - Serotonin pharmacology in the gastrointestinal tract: a review. AB - Serotonin (5-hydroxytryptamine or 5-HT) plays a critical physiological role in the regulation of gastrointestinal (GI) function. 5-HT dysfunction may also be involved in the pathophysiology of a number of functional GI disorders, such as chronic constipation, irritable bowel syndrome and functional dyspepsia. This article describes the role of 5-HT in the enteric nervous system (ENS) of the mammalian GI tract and the receptors with which it interacts. Existing serotonergic therapies that have proven effective in the treatment of GI functional disorders and the potential of drugs currently in development are also highlighted. Advances in our understanding of the physiological and pathophysiological roles of 5-HT in the ENS and the identification of selective receptor ligands bodes well for the future development of more efficacious therapies for patients with functional GI disorders. PMID- 18398607 TI - Differential responses of cellular immunity in patients undergoing neoadjuvant therapy followed by surgery for carcinoma of the oesophagus. AB - BACKGROUND: To compare immune responses following neoadjuvant chemoradiation therapy in combination with hyperthermia plus surgery to those induced by surgery alone in patients with oesophageal cancer. METHODS: Thirty-two patients with histopathologically proven oesophageal cancer, scheduled for potentially curative transhiatal or transthoracic oesophagectomy with (neo, n = 20) or without (control, n = 12) neoadjuvant thermochemoradiation therapy (ThCR) were included. Peripheral blood samples were obtained before ThCR, after 2 weeks of ThCR, 1 day before surgery, on postoperative days 1, 3, 7, and 6 weeks after surgery, for white blood cell counts, lymphocyte subsets and T helper type 1 (Th1) and type 2 (Th2) lymphocyte responses. RESULTS: Neo patients showed a significant decrease in granulocytes and lymphocyte subsets, and T cell cytokines after 2 weeks of ThCR. Only CD8+ (cytotoxic) T cells recovered after ThCR to reach normal levels prior to surgery. In contrast, CD4+ T (helper) cells, and NK- and B cells in neo patients did not recover prior to surgery (all P < 0.05). Oesophagectomy induced a significant increase in granulocytes and a decrease in lymphocytes (and subsets). Only those subsets that had not recovered after ThCR (CD4+ T cells, NK and B cells but not CD8+ T cells), were significantly lower (all P < 0.05) during the entire postoperative study period. Postoperatively, the stimulated cytokine production capacity of Th1 and Th2 cells, corrected for number of T cells, was not significantly different between the groups. CONCLUSION: Neoadjuvant thermochemoradiation for oesophageal cancer caused significant disturbances of host cellular immunity with reduced T, NK and B cell counts, and differential recovery of cytotoxic and helper T cells leading to prolonged T cell imbalance that extends beyond the time of surgery. The functional and anti-tumour consequences of this immunodisturbance need further investigation, as recovery of T helper cytokine production towards surgery was less impaired than T helper cell counts. PMID- 18398608 TI - Treatment outcomes of oxaliplatin, 5-FU, and leucovorin as salvage therapy for patients with advanced or metastatic gastric cancer: a retrospective analysis. AB - PURPOSE: We performed a single-institution retrospective study to evaluate the efficacy and toxicities of oxaliplatin, 5-fluorouracil (5-FU), leucovorin (LV) combination chemotherapy as salvage treatment in patients with metastatic or advanced gastric cancer. METHODS: Sixty-two patients with advanced gastric cancer previously treated were eligible for the study. Patients received oxaliplatin 100 mg/m(2) and LV 100 mg/m(2) (2-h intravenous infusion) followed by 5-FU 2,400 mg/m(2) (46-h continuous infusion) every 2 weeks, and responses were assessed after every three cycles. RESULTS: Fifty-nine out of 62 patients were assessable for response. Among them, 46 patients had previously been treated with cisplatin based chemotherapy. Patients had a median age of 57 years (range 32-76 years), 72.6% had an Eastern Cooperative Oncology Group performance status of 0 or 1. Total 296 courses of chemotherapy were administered as second-line (67.7%) or third-line (27.4%), and the median courses per patient was three cycles. Out of 59 evaluable patients, 14 partial responses were observed (overall response rate, 22.6%). Stable disease was observed in 22 patients (35.5%), and progressive disease in 23 patients (37.1%). The median response duration, time to progression, and overall survival were 2.3, 3.0, and 8.0 months, respectively. The major toxicities were neutropenia, mucositis, and peripheral neuropathy. Grade 3 or 4 hematologic toxicities included neutropenia in nine patients (14.5%) and thrombocytopenia in one patient (1.6%). Other grade 3 or 4 toxicities included mucositis in one patient (1.6%) and vomiting in two patients (3.2%). Grade 1 or 2 peripheral neuropathy were observed in 18 patients (29.0%), however there were no cases of grade 3 or 4 peripheral neuropathy and no treatment related deaths. CONCLUSION: The combination of oxaliplatin, 5-FU and LV was effective and safe salvage chemotherapy in advanced gastric cancer patients. PMID- 18398609 TI - Inhibition of cytidine deaminase by zebularine enhances the antineoplastic action of 5-aza-2'-deoxycytidine. AB - Cytidine (CR) deaminase is a key enzyme in the catabolism of cytosine nucleoside analogues, since their deamination results in a loss of their pharmacological activity. In this report we have investigated the importance of CR deaminase with respect to the antineoplastic action of inhibitors of DNA methylation, 5-aza-2' deoxycytidine (5-AZA-CdR) and zebularine. Zebularine has a dual mechanism of action, since it can also inhibit CR deaminase. The objective of our study was to investigate the importance of zebularine as an inhibitor of CR deaminase with respect to the antineoplastic action of 5-AZA-CdR. Using an in vitro clonogenic assay, we investigated the antineoplastic action of 5-AZA-CdR and zebularine, alone and in combination on wild type 3T3 murine fibroblasts and corresponding V5 cells transduced with CR deaminase gene to express a very high level of CR deaminase activity. The V5 cells were much less sensitive to 5-AZA-CdR than the wild type 3T3 cells. The addition of zebularine significantly enhanced the antineoplastic action of 5-AZA-CdR on V5 cells, but not 3T3 cells. Enzymatic analysis on CR deaminase purified from the V5 cells showed that zebularine is a competitive inhibitor of the deamination of 5-AZA-CdR. These in vitro observations are in accord with our in vivo study in mice with L1210 leukemia, which showed that zebularine increased the antileukemic activity of 5-AZA-CdR. Pharmacokinetic analysis also showed that zebularine increased the plasma level of 5-AZA-CdR during an i.v. infusion in mice. Our results indicate that the major mechanism by which zebularine enhances the antineoplastic action of 5-AZA-CdR is by inhibition of CR deaminase. These findings provide a rationale to investigate 5-AZA-CdR in combination with zebularine in patients with advanced leukemia. PMID- 18398610 TI - Combination therapy with docetaxel and low dose of cisplatin in elderly patients with advanced non-small cell lung cancer: multicenter phase II study. AB - PURPOSE: To determine the efficacy and safety of the combination therapy with docetaxel and cisplatin (CDDP) at low doses in elderly patients with advanced NSCLC. PATIENTS AND METHODS: A total of 42 patients aged > or =70 years with previously untreated advanced NSCLC received docetaxel 75 mg/m(2) plus CDDP 50 mg/m(2) on day 1. The regimen was repeated every 21 days. Patients received a minimum of three courses unless progressive disease was detected. RESULTS: By intent-to-treat analysis, the overall response rate was 31% (95% CI, 17.8-47.2%). A total of 18 patients (43%) had stable disease and 11 (26%) progressed. Median time to progression was 5.2 months. Overall median survival was 8.9 months, with 1-year actuarial survival rate of 41%. Eastern Cooperative Oncology Group performance status was improved in 18 patients (43%). The chemotherapy regimen was well tolerated. A total of 11 patients (26%) had grade 3/4 adverse events: 7 (17%) neutropenia (one of them was diagnosed with febrile neutropenia), 3 (7%) asthenia, 3 (7%) nausea/vomiting, 1 (2%) diarrhea, 1 (2%) thrombocytopenia and 1 (2%) neurotoxicity. No death due to toxicity was seen. CONCLUSION: The combination of low-dose CDDP and docetaxel for elderly patients with advanced NSCLC is an efficient and well-tolerated chemotherapeutic approach. PMID- 18398611 TI - A novel phenotypic method to determine fludarabine triphosphate accumulation in T lymphocytes from hematopoietic cell transplantation patients. AB - PURPOSE: Fludarabine is an integral anticancer agent for patients with chronic lymphocytic leukemia (CLL) and those receiving conditioning regimens prior to allogeneic hematopoietic cell transplantation (HCT). An individual's response to fludarabine may be influenced by the amount of CD4(+) and CD8(+) T-lymphocyte suppression. Fludarabine undergoes cellular uptake and activation to form the cytotoxic metabolite, fludarabine triphosphate (F-ara-ATP). METHODS: We have previously developed a highly sensitive LC-MS method to quantitate intracellular F-ara-ATP concentrations in a leukemic cell line. However, quantitation of F-ara ATP concentrations within CD4(+) and CD8(+) T-lymphocytes from pharmacokinetic blood samples obtained from patients receiving fludarabine therapy is not feasible because of the limited number of T-lymphocytes that can be isolated from each blood sample. Thus, we sought to determine F-ara-ATP accumulation after ex vivo exposure of freshly isolated human CD4(+) or CD8(+) T-lymphocytes to fludarabine. The method was optimized in T-lymphocytes obtained from healthy volunteers, and proved to be a feasible method to determine F-ara-ATP accumulation in patients undergoing HCT. RESULTS: Considerable variability was observed in F-ara-ATP accumulation in HCT patients (10.5- and 12.5-fold in CD4(+) and CD8(+) cells, respectively), compared to healthy volunteers (1.6- and 1.9 fold in CD4(+) and CD8(+) cells, respectively). Larger variability was also observed in gene expression of transporters and enzymes involved in F-ara-ATP accumulation in HCT patients; however, F-ara-ATP accumulation was not correlated with gene expression, which is in agreement with previous studies. CONCLUSIONS: The quantitation of F-ara-ATP accumulation in T-lymphocytes provides a novel tool to evaluate patient sensitivity to fludarabine. This tool can be used in future studies to evaluate whether intracellular F-ara-ATP accumulation is associated with efficacy and/or toxicity in patients receiving fludarabine. PMID- 18398612 TI - Valproic acid as a therapeutic agent for head and neck squamous cell carcinomas. AB - PURPOSES: Here we investigate if valproic acid (VA) can enhance the efficacy of commonly used therapies for head and neck squamous cell carcinomas (HNSCC) and the molecular mechanisms that may be related to its anticancer effects. METHODS: Proliferation and viability of distinct cell types subjected to VA treatment alone or in combination regimens were measured through BrdU incorporation and LDH release, respectively. Molecular markers compatible with histone deacetylase inhibitory activity of VA were assessed through western blots assays in lysates obtained from cultured cells and tumour biopsies. RESULTS: Treatment of all cell types with VA resulted in a dose-dependent increase in histone H3 acetylation and p21 expression, as well as dose-dependent cytostasis. In contrast, the cytotoxic response to VA was variable and did not correlate with cytostasis, histone acetylation or p21 induction. The variability in response to VA was also observed in tumour biopsy samples collected from patients prior to and following a 1 week oral course of VA. In addition, we found that a combination of a clinically achievable concentration of VA plus cisplatin caused a threefold to sevenfold increase in cisplatin cytotoxicity in vitro. CONCLUSIONS: VA acts as a histone deacetylase inhibitor (HDI) in SCC cells and normal human keratinocytes (HKs), potentiates the cytotoxic effect of cisplatin in SCC cell lines and decreases the viability of SCC cells as opposed to HKs. Taken together, the results provide initial evidence that VA might be a valuable drug in the development of better therapeutic regimens for HNSCC. PMID- 18398613 TI - A phase II study of pegylated-camptothecin (pegamotecan) in the treatment of locally advanced and metastatic gastric and gastro-oesophageal junction adenocarcinoma. AB - PURPOSE: Combination chemotherapy results in a significant survival advantage in patients with advanced gastric cancer compared to best supportive care. Nevertheless, the prognosis remains poor with a median survival of 8-10 months. Topoisomerase-I inhibitors such as irinotecan have activity in advanced gastric cancer. Pegamotecan may offer significant advantages over other topoisomerase-I inhibitors due to its prolonged circulating half-life, tolerability and passive tumour accumulation. PATIENTS AND METHODS: This was a non-randomised, multi centre, two-step Fleming design phase II study. Eligible patients with locally advanced (inoperable) or metastatic gastric or gastro-oesophageal adenocarcinoma, with measurable disease, ECOG performance status < or =2, with adequate haematological, renal and hepatic function, who had received < or =1 prior chemotherapy regimen for advanced disease, were treated with 7,000 mg/m(2) of pegamotecan as a 1-h infusion every 21 days until disease progression or unacceptable toxicity. The primary efficacy measure was the objective response rate. RESULTS: Five of the 35 patients recruited into this study had a partial response (14.3%), with a median time to progression of 11.9 weeks (95% CI: 6.6, 13.1), and median overall survival of 38.1 weeks (95% CI: 29.0, 47.3). Grade 3/4 toxicities included neutropenia in 6 (17.1%) patients, thrombocytopenia in 4 (11.4%), fatigue in 8 (22.9%), nausea in 6 (17%), vomiting in 6 (17%) and anorexia in 4 (11.4%) patients. There were no episodes of febrile neutropenia and no toxic deaths. CONCLUSIONS: Pegamotecan has activity in this patient population and was generally well-tolerated. The favourable rate of haematological toxicities and diarrhoea compared with irinotecan in similar studies suggests that pegamotecan could be combined with other active agents in further studies in this disease. PMID- 18398614 TI - A phase II study of S-1 in gemcitabine-refractory metastatic pancreatic cancer. AB - PURPOSE: Gemcitabine monotherapy or gemcitabine-containing combination chemotherapy is the standard first-line therapy for advanced pancreatic cancer. After disease progression, there is no standard regimen available. In a previous phase II trial, S-1 has been reported to show considerable efficacy, achieving a response rate of 37.5% in chemo-naive patients with pancreatic cancer. This study evaluated the efficacy and toxicity of S-1 in patients with gemcitabine refractory metastatic pancreatic cancer. METHODS: Eligibility criteria were histologically proven pancreatic adenocarcinoma with confirmation of progressive disease while receiving gemcitabine-based first-line chemotherapy, 20-74 years of age, Karnofsky performance status of 80-100 points, with measurable metastatic lesions, adequate hematological, renal and liver functions, and written informed consent. S-1 was administered orally at 40 mg/m(2) twice daily for 28 days with a rest period of 14 days as one course. Administration was repeated until the appearance of disease progression or unacceptable toxicity. The primary endpoint of this study was an objective response, and secondary endpoints included toxicity, progression-free survival (PFS) and overall survival, as well as clinical benefit response in symptomatic patients. RESULTS: Forty patients from two institutions were enrolled between September 2004 and November 2005. The most common adverse reactions were fatigue and anorexia, although most of those adverse reactions were tolerable and reversible. One patient developed grade 3 pneumonitis without neutropenia and recovered with appropriate antibiotic treatment. Although no complete response was seen, partial response was obtained in six patients (15, 95% confidence interval, 3.9-26%). Stable disease was noted in 17 patients (43%), and progressive disease in 15 patients (38%). Out of 19 evaluable patients, a clinical benefit response was observed in four patients (21%). The median PFS was 2.0 months, and the median survival time was 4.5 months with a 1-year survival rate of 14.1%. CONCLUSION: S-1 as monotherapy had marginal anti-tumor activity with tolerable toxicity in patients with gemcitabine refractory metastatic pancreatic cancer. PMID- 18398615 TI - Population pharmacokinetics of imatinib mesylate and its metabolite in children and young adults. AB - BACKGROUND: Imatinib mesylate (Gleevec) is a small molecule tyrosine kinase inhibitor approved for use in the management of chronic myeloid leukemia in adults and children and in gastrointestinal stromal tumors in adults. Population pharmacokinetic (PPK) studies evaluating the effect of population covariates on the pharmacokinetics of imatinib and its active metabolite have been developed in adults with chronic myeloid leukemia (CML) and gastrointestinal stromal tumor (GIST). However, this still remains to be described in children. PURPOSE: The objectives of the analysis were to develop a PPK model of imatinib and its active metabolite, CGP74588, to describe exposure in children and young adults and to identify covariates that are predictors of variability in disposition. METHODS: Plasma concentrations from 26 subjects with Philadelphia (Ph+) leukemia (Phase I study) and 15 subjects with refractory solid tumors (Phase II study), who received oral imatinib at doses ranging from 260 to 570 mg/m(2), were available for the PPK analysis in NONMEM. Blood samples were drawn prior to dosing and over 24-48 h on days 1 and 8 of the studies. Covariates studied included weight, age, albumin, alanine aminotransferase and the study population. RESULTS: The pharmacokinetics of imatinib and CGP 74588 were well described by one and two compartment models, respectively. Total body weight was the only covariate found to significantly affect Cl/F and V/F. The final imatinib-CGP 74588 model is summarized as follows: CL/F (imatinib) (L/h) = 10.8 x (WT/70)(0.75), V/F (imatinib) (L) = 284 x (WT/70) and D1(duration of zero order absorption,imatinib) (h) = 1.67 and CL/F (CGP 74588) (L/h) = 9.65 x (WT/70)(0.75), V1/F (CGP 74588) (L) = 11.6 x (WT/70), Q (CGP 74588) (L/h) = 2.9 x (WT/70)(0.75) and V2/F (CGP 74588) (L) = 256*(WT/70). Model evaluation indicated that the final model was robust and satisfactory. CONCLUSIONS: Current imatinib dosing guidelines in pediatrics is based on the achievement of exposures consistent with doses known to be safe and efficacious in adults. Dose adjustments in children are guided empirically by the observance of drug-related toxicities. While, the pharmacokinetics of imatinib and its active metabolite, CGP 74588 in children are consistent with prior knowledge in adults, the model will form the basis to support the design of future trials, particularly with a view to managing toxicities and exploring dosing in this population. PMID- 18398616 TI - Severe hemorrhagic cystitis associated with prolonged oral cyclophosphamide therapy: case report and literature review. AB - Severe hemorrhagic cystitis associated with oral cyclophosphamide (CYP) therapy has rarely been reported in the past 20 years, probably because this condition has largely disappeared because of the use of shorter courses of CYP, either oral or IV. Herein, we describe a patient who received 309 g of oral CYP over a 4-year period to treat Wegener's granulomatosis (WG) that initially involved brain, lung, and kidney. She came under our care for the first time when she presented with a one-day history of oliguria and passing blood clots. Severe hemorrhagic cystitis was present. It eventually required cystectomy. Despite her massive CYP exposure her kidney biopsy showed acute crescentic glomerulonephritis. She survived her acute illness only to die 2 months later of acute leukemia. This patient is a reminder that severe hemorrhagic cystitis from oral CYP still exists, and that WG can be resistant to even toxic doses of oral CYP. Alternative therapies are discussed. PMID- 18398617 TI - Two-year outcomes in primary THA in obese male veterans administration medical center patients. AB - The objective of this study was to examine the influence of obesity on the rate of infection, dislocation, and overall complications in a group of male patients undergoing primary total hip arthroplasty (THA). The study involved a retrospective review of the association of body mass index (BMI) and dislocation, infection, and complications on 255 consecutive male patients who received a THA. BMI of 40 and greater had significantly more dislocations and approached significance for infections. The present study suggests that the risk of infection and dislocation increases only in highly obese males (BMI > 40) in contrast to other studies that have reported a higher risk infection, dislocation, and complications with a BMI of obesity in females (BMI > 30). PMID- 18398619 TI - [CCL2/MCP1: a novel target in systemic lupus erythematosus and lupus nephritis]. AB - Cytokine blockade, a valid therapeutic concept, is not established in lupus nephritis as yet. In lupus nephritis CCL2/MCP-1 and its chemokine receptor CCR2 are of interest because CCL2/CCR2 mediate the recruitment of macrophages and T cells in the nephritic kidney. Lupus nephritis is markedly attenuated in CCL2- or CCR2-deficient autoimmune mice. Epidemiological studies addressing mutations in the CCL2 gene support the hypothesis that CCL2 mediates renal inflammation. Meanwhile experimental studies have shown that several classes of CCL2 antagonists can control established lupus nephritis. Interestingly, therapeutic CCL2 blockade does not affect the autoimmune lymphoproliferative syndrome and the production of lupus autoantibodies. This article briefly summarizes the potential role of therapeutic CCL2 blockade in lupus nephritis. PMID- 18398620 TI - The effect of acute hyperglycemia on retinal thickness and ocular refraction in healthy subjects. PMID- 18398621 TI - Standard-radical vs. function-preserving surgery of benign nodular goiter: a sonographic and biochemical 10-year follow-up study. AB - BACKGROUND: The necessary extent of thyroid resection in benign nodular goiter is under debate. The aim of our study was to compare the long-term outcome of different thyroid resection modes with special interest in the incidence of recurrent nodules and the use of oral thyroid hormone medication. MATERIALS AND METHODS: We performed a follow-up examination of 109 patients (23 men and 86 women) having been operated for benign nodular goiter at our department 10 years ago. Unilateral resections and function-preserving resections of at least one thyroid lobe were classified as function-preserving (FP). Total thyroidectomy, Dunhill's operation and bilateral subtotal thyroidectomy were rated as standard radical (STR). On follow-up, we recorded current oral thyroid hormone medication, thyroid function tests and ultrasound of the neck. RESULTS: Seventy-three patients had FP resection (67%), while 36 were STR-operated (33%). The subsequent medical treatment was performed by dedicated endocrinologists (n = 19), internists (n = 11) or primary-care physicians (n = 59). Twenty patients had no medical attendance. Recurrent nodules were found in 13 cases in the FP group (18.6%) vs. 3 cases in the STR group (2.5%; p < 0.001). In both groups, about 80% of patients used thyroid hormone medication 10 years after operation. CONCLUSION: There was no advantage in thyroid function tests nor lesser medication in the FP group. The risk for recurrent nodules was significantly higher in the FP than in the STR-operated patients. PMID- 18398622 TI - Impact of rapamycin on liver regeneration. AB - The remarkable capacity of the liver to regenerate after injury and the prospects of organ self-renewal have attracted much interest in the understanding and modulation of the underlying molecular events. We investigated the effect of mammalian target of rapamycin (mTOR) inhibitor rapamycin (RAPA) on liver by correlating intravital microscopy, immunohistochemistry, and reverse transcriptase polymerase chain reaction in a rat model of 2/3 hepatectomy. RAPA significantly retarded proliferation of hepatocytes, endothelial cells, and hepatic stellate cells (HSCs) mostly between days 2 and 4 after hepatectomy and downregulated major cytokines and growth factors (tumor necrosis factor alpha, hepatocyte growth factor, platelet-derived growth factor, platelet-derived growth factor receptor, insulin-like growth factor-1, transforming growth factor beta 1) important for liver regeneration. These effects were almost absent at later time points. RAPA also had a transient, but broad effect on angiogenesis, and impaired sinusoidal density as well as mRNA levels of vascular endothelial growth factor, vascular endothelial growth factor receptor 1, vascular endothelial growth factor receptor 2, and angiopoietin-1. Activation of HSC was also transiently suppressed as observed by smooth muscle protein 1 alpha protein expression and intercellular adhesion molecule-1 mRNA levels. The rate of apoptosis in liver was significantly increased by RAPA between day 3 and day 7. The effect of RAPA on liver repair, angiogenesis, and HSC activation is confined to the phase of active cell proliferation. This transient effect might allow further exploration of mTOR inhibitors in clinical situations that involve liver regeneration, and seems to have implications beyond immunosuppression. PMID- 18398623 TI - Umbilical artery blood S100beta protein: a tool for the early identification of neonatal hypoxic-ischemic encephalopathy. AB - Neuroprotective interventions in neonatal hypoxic-ischemic encephalopathy (HIE) require early indicators of brain damage to initiate therapy. In order to find a reliable, rapid, and simple method to identify infants at risk for this disorder, 40 infants with asphyxia were selected as the observation group (HIE group) and 25 normal-term infants as the control group. S100beta protein concentration and gas analysis of the umbilical cord artery blood of all infants were determined. We found that the S100beta protein levels of the HIE group (1.98 microg/L) were higher than those of the control group (1.05 microg/L, p<0.05), and there were significant differences between the mild HIE group (1.72 microg/L) and the moderate or severe HIE groups (3.61 microg/L, p<0.05). An S100beta protein concentration cutoff level of 2.02 microg/L had a sensitivity of 86.7% and a specificity of 88.0% for predicting the development of moderate or severe HIE. The blood gas parameters of umbilical artery blood, such as pH, carbon dioxide tension, and base excess, were significantly different in the HIE group compared to the control group (all p<0.001), but there were no differences between the mild HIE group and the moderate or severe HIE groups. On the basis of clinical manifestations of asphyxiated neonates, detecting the S100beta protein levels in the umbilical artery blood may be of important value in the early diagnosis and grading of HIE. PMID- 18398624 TI - Pyruvate dehydrogenase deficiency: identification of a novel mutation in the PDHA1 gene which responds to amino acid supplementation. AB - The pyruvate dehydrogenase complex (PDHc) is an intramitochondrial multienzyme system, which plays a key role in aerobic glucose metabolism by catalysing the oxidative decarboxylation of pyruvate to acetyl-CoA. Genetic defects in the PDHc lead to lactic acidemia and neurological abnormalities. In the majority of the cases, the defect appears to reside in the E(1)alpha subunit, the first catalytic component of the complex. The report is on a 6-year-old Portuguese boy with mild neurological involvement and low PDHc activity with absence of E1alpha on immunoblotting analysis. Molecular studies showed a novel and "de novo" mutation in the PDHA1 gene, R253G. Treatment with arginine aspartate showed complete clinical and biochemical recovery. We hypothesise that arginine aspartate acts as a chemical or pharmacological chaperone, and suggest amino acid supplementation as a possible therapy in PDHA1 mutations with mild phenotypes. CONCLUSION: our results encourage the use of amino acid supplementation to overcome the metabolic/biochemical changes induced by PDHA1 gene specific mutations associated with mild PDHc phenotypes. PMID- 18398625 TI - Pseudolipoma in an 18-month-old Caucasian girl: no trauma reported. AB - We report a pseudolipoma in an 18-month-old Caucasian girl without history of trauma. Clinical and histological findings are discussed and the literature is reviewed. PMID- 18398626 TI - Differential diagnosis of oocysts of Hammondia-like organisms of dogs and cats by PCR-RFLP analysis of 70-kilodalton heat shock protein (HSP70) gene. AB - Organisms of the genera Toxoplasma, Hammondia and Neospora, the Hammondia-like organisms, are closely related coccidian with similarly sized oocysts. Therefore, a diagnosis based on microscopy of oocysts in feces is not a method of choice for species identification of these important parasites. In this paper, we present a polymerase chain reaction coupled with restriction fragment length polymorphism (PCR-RFLP) method to differentially diagnose oocysts of Toxoplasma gondii from oocyst of Hammondia hammondi. Another PCR-RFLP was designed to differentiate oocysts of Hammondia heydorni from oocysts of Neospora spp. Both PCR-RFLP are based on nucleotide sequences of the Hsp70 coding gene. In conclusion, we presented two alternative molecular diagnostic assays that can be successfully applied for the differentiation of oocysts of Hammondia-like organisms shed by felids and canids. PMID- 18398628 TI - A three-dimensional elastic plastic damage constitutive law for bone tissue. AB - Motivated by mechanical analysis of bones and bone-implant systems, a 3D constitutive law describing the macroscopic mechanical behaviour of both cortical and trabecular bone in cyclic (not fatigue) overloads is developed. The proposed model which mathematical formulation is established within the framework of generalized standard materials accounts for three distinct material evolution modes where elastic, plastic and damage aspects are closely related. The anisotropic elasticity of bone is described by a morphology-based model and distinct damage behaviour in tension and compression by a halfspacewise generalized Hill criterion. The plastic criterion is based on the intact elastic compliance tensor. The algorithm applies three distinct projections based on the relationship between the internal variables and criteria. Their respective consistent tangent operators are presented. Numerical resolutions of several boundary value problems and a biomechanical application are presented to illustrate the potential of the constitutive model and demonstrate the expected quadratic convergence of the algorithm. PMID- 18398627 TI - The effect of different dental treatment strategies on the oral health of children: a longitudinal randomised controlled trial. AB - The aim of the present study is to verify which strategy is the most effective in the treatment of dental decay of the deciduous dentition in a moderate to high caries child population under remote field conditions. This study was carried out in the rain forest of Suriname. Three hundred and eighty schoolchildren, mean age 6.1 years (SD 0.5, range 5.1-7.1 years), were randomly assigned to four different groups: full dental treatment, only extractions, only restorations (ART) and no treatment. Parameters for oral health were defined as caries prevalence (dmft), caries increment, sequela to dental caries and dental pain. Restorative dental care of the primary dentition, by means of ART, resulted in a caries increment from a dmft of 5.48 (SD 3.2) at baseline to 6.35 (SD 2.6) after 2 years (p < 0.001). Extensive dental treatment, performing only extractions, or no treatment did not render significant changes in the caries prevalence of children (p > 0.05). Full dental treatment should be the strategy of choice whenever oral health care programmes are developed. However, when priorities are required due to situational, practical or economical reasons, extraction of severely decayed teeth is an effective treatment strategy. PMID- 18398630 TI - Prevalence of probable obstructive sleep apnea risk and severity in a population of dental patients. AB - Obstructive sleep apnea is a commonly undiagnosed chronic disease. While dentists represent an important resource for identifying people at risk for primary snoring and sleep apnea, less than 50% of dentists are capable of identifying the common signs and symptoms of sleep disordered breathing. The aim of this study is to assess the prevalence of probable obstructive sleep apnea/sleep disordered breathing and symptoms associated with this condition in a population of dental patients using a validated questionnaire and software that could be administered in a dental office. A retrospective analysis conducted at two dental practices using questionnaire responses obtained from 175 men and 156 women, and sleep study data obtained in the patient's homes from 75 men and 30 women with a portable recorder. Forty-six percent of the men and 19% of the women reported snoring frequently or always. Of the 67% of the men and 28% of the women identified as having a high pre-test probability (high risk) of having at least mild sleep apnea, over 33% of the men and 6% of the women surveyed were predicted to have moderate or severe sleep apnea. In a subgroup of 105 patients classified at high risk who completed an overnight sleep study, 96% had an apnea hypopnea index (AHI) greater than five events per hour. Seventy percent of those predicted to have moderate or severe OSA by questionnaire had an AHI greater than 20. All patients previously diagnosed with sleep apnea were correctly classified at high risk by ARES questionnaire. There was a high concordance between the predicted OSA risk and the degree of sleep disordered breathing. The high prevalence of undiagnosed sleep apnea in dental patients suggests that dentists could provide a valuable service to their patients by incorporating sleep apnea screening and treatment into their practice. Those who practice sedation dentistry should consider additional precautions when managing patients with risk of sleep apnea. PMID- 18398629 TI - Hemostatic implications of endothelial cell apoptosis in obstructive sleep apnea. AB - Patients with obstructive sleep apnea (OSA) are at increased risk of atherothrombosis independent of the Framingham risk factors. Studies on hemostasis factors in OSA are scarce and inconsistent. We sought to understand the variation in atherothrombotic propensity as a function of apoptotic circulating endothelial cells (CECs) in OSA by investigating the relationship between CEC apoptosis and plasma levels of hemostatic factors tissue factor (TF) and von Willebrand Factor (vWF) in apneic subjects. Apoptotic CECs were detected by flow cytometry in 35 male subjects free of cardiovascular diseases (AHI range 8-43) and 12 healthy male controls (AHI range 2-5) before and after 8 weeks of nasal continuous positive airway pressure (nCPAP). Quantitative determination of TF and vWF was performed using an enzyme-linked immunosorbent assay (ELISA) kit. The mean levels of TF (66.78 +/- 41.59 pg/ml) and vWF (189.70 +/- 69.24 IU/dl) were significantly higher in OSA patients compared with those in healthy subjects (42.83 +/- 14.18 pg/ml; and 124.48 +/- 31.43 IU/dl). Apoptotic CECs were elevated in patients with OSA and correlated strongly with TF and vWF levels (p = 0.02 and p < 0.001; respectively). There were no correlations between TF, vWF and apnea hypopnea index, or arousal index. Only the percentage of time spent <90% oxygen saturation was inversely associated with TF (r = 0.38; p = 0.02). Following nCPAP therapy, there was significant decrease in TF levels that correlated with decrease in apoptotic CECs. In patients with OSA, increased prothrombotic factors are strongly determined by apoptotic CECs. Treatment with nCPAP may alleviate the coagulation propensity. PMID- 18398631 TI - Health status of the elderly population among four primitive tribes of Orissa, India: a clinico-epidemiological study. AB - Primitive tribal groups (PTGs) are the most marginalised and vulnerable communities in India. Clinico-epidemiological studies on morbidity patterns among the elderly primitive tribe members are essential to recommend special intervention programmes to improve the health of the elderly in these communities. A community-based cross-sectional study was carried out among the elderly populations of four different PTGs, namely Langia Saora (LS), Paudi Bhuiyan (PB), Kutia Kondh (KK) and Dongria Kondh (DK) living in the forests of Orissa, India. Clinical and anthropometric data were collected using standard methods and haemoglobin was estimated by the cyanomethaemoglobin method. The average number of illnesses per person was 3.0. Common disabilities like vision and hearing impairment and mobility-related problems were found in considerable numbers. Gastrointestinal problems like acid peptic disease were found among 2.6% to 20% of cases. Non-specific fever was marked in 10.2% to 24.2% of individuals. The iodine deficiency disorder, namely goitre, was found among 4.2% to 6.0% of individuals. Diseases of the respiratory tract, like upper and lower respiratory tract infection, asthma, tuberculosis and leprosy, were found in small numbers. The prevalence of hypertension among males and females was 31.8% and 42.2%, respectively. The LS had the highest prevalence of hypertension (63% among men and 68% among women). With regard to anaemia status, severe anaemia was marked in 70% of males and 76.7% of females in the LS, while in other groups the prevalence of severe anaemia ranged from 15% to 33%. Although the prevalence of severe anaemia in other tribal communities is lower than in the LS, mild to moderate anaemia was found to range from 60% to 80%. The present study revealed a high prevalence of physical disabilities with both non-communicable as well as communicable diseases among the elderly primitive tribal members. This warrants the implementation of a special health care strategy to reduce suffering at this crucial age and improve quality of life. PMID- 18398632 TI - [Practical guideline for nutritional care in geriatric institutions]. AB - Malnutrition is widespread in geriatric patients, but often unrecognized and untreated. The main barriers against implementation of adequate nutritional care are seen in low nutritional awareness, lack of knowledge, interest and responsibilities but also in established procedures and lack of standard protocols for nutritional screening and therapy. It is, thus, the aim of the present publication to provide a comprehensive, practice oriented guideline for quality assurance of nutritional care of elderly people in geriatric institutions. The guideline includes routine assessment of individual nutritional status as well as the planning and implementation of appropriate interventions and measurement of results. The guideline is aimed at the prevention and adequate treatment of weight loss and malnutrition and is composed of the following sections: 1) screening, 2) assessment, 3) definition of aims, 4) intervention, 5) monitoring with adaptation of aims and interventions, 6) planning of nutritional care after discharge. Appropriate interventions aim, on the one hand, at the elimination of underlying causes of malnutrition and, on the other hand, at adequate and sufficient dietary intake to meet the requirements. Dietetic, nursing and medical actions should be implemented in a coordinated manner in order to use all options to achieve the best nutritional care for each patient. As a prerequisite for high quality individual nutritional care, several basic conditions have to be established, e.g. appropriate supply of food and beverages to meet patient needs, well-organized caring processes during meals, willingness of all involved persons for interdisciplinary teamwork, and well-regulated responsibilities. This general guideline has to be adapted to the local conditions of each institution and consistently put into practice. For implementation of the guideline, a nutrition team with members of all relevant professions should be organized with regular meetings. In the long run, increasing nutritional awareness and established routines for adequate nutritional screening and therapy will enable high quality nutritional care of geriatric patients with manageable charges. PMID- 18398633 TI - [Social inequality and health care among the aged]. AB - In this article the association between social inequality and selected aspects of health care among the aged is analysed. Analyses are based on German data (release 1) of the 'Survey of Health, Ageing and Retirement in Europe' (SHARE) in 2004. Data from 1921 respondents aged 50 years or more are analysed. Three indicators of social inequality are used (education, income, and financial assets). In terms of health care, indicators of geriatric assessment by the general practitioner of the respondent (questions about balance, physical exercise, and drugs as well as examination of balance and weight control) and secondary prevention (mammogram, endoscopic examination of colon, eye examination) are included. Results of cross-tabs and logistic regression analyses show that geriatric assessment is less comprehensive among people in a comparatively high socioeconomic position. On the other hand, people in a higher socioeconomic position use screening examinations more often than those in a lower position. Inconsistency of results indicates the necessity to distinguish different areas of health care when analysing social inequalities. Moreover, results indicate that no simple answer can be given to the question whether and to what extent social inequalities in health among older people can be explained by inequalities in medical care. PMID- 18398634 TI - Autonomic responses to physiological stressors in women with type 2 diabetes. AB - OBJECTIVE: To compare autonomic function, measured during handgrip (HG) and cold pressor (CP), between obese with and without type 2 diabetes and non-obese women in fasting and post-glucose load states. METHODS: Twelve obese women with type 2 diabetes (50 +/- 1 years), 15 obese women without type 2 diabetes (48 +/- 2 years), and 12 non-obese women (49 +/- 2 years) participated in this study. Heart rate variability (HRV) was determined during autonomic function tests, conducted in both the fasting state and after a glucose challenge (oral glucose tolerance test-OGTT). RESULTS: Obese women with and without diabetes and non-obese women responded similarly fasted and post-glucose challenge, such that in the fasted state low frequency power normalized (LF(nu)) to total power (TP), log transformed (Ln) low frequency to high frequency ratio (LnLF/HF) and heart rate (HR) significantly increased with the autonomic functional tasks (P < 0.05), whereas HF(nu) significantly decreased with the tasks (P < 0.05). Handgrip elicited a lower LnTP and a higher HR (P < 0.05) when compared to CP in the fasted state. In the glucose challenged state LF(nu), LnLF/HF and HR increased (P < 0.05) and HF(nu) significantly decreased (P < 0.05). INTERPRETATION: Results of autonomic testing did not differ between obese women, with and without diabetes, and non-obese women. The HG test elicited a greater reduction in HRV total power compared to the CP. This suggests that HG may be more useful when examining autonomic function in women with complicated diabetes. PMID- 18398635 TI - The quest for the unholy grail. PMID- 18398636 TI - Changes in forest area along stream networks in an agricultural catchment of the Great Barrier Reef lagoon. AB - Scenes from the series of multispectral sensors on the Landsat satellites were used to map recent changes (between 1972 and 2004) in forest cover within and adjacent to stream networks of an intensively farmed region of the southern Great Barrier Reef catchment (Australia). Unsupervised ISODATA classifications of Tasseled-Cap transformed data (at 57 m ground resolution) mapped forest and cleared areas within 150 m of Pisoneer catchment waterways with 72.2% overall accuracy (K(hat) = 0.469), when adjusted for the size of each class. Although the user's accuracy was higher for the forest class (82.1 +/- 8.4% at alpha = 0.05), large errors of commission (34.2 +/- 8.3%) substantially affected map accuracy for the cleared class. The main reasons for misclassification include: (1) failure to discriminate narrowly vegetated riparian strips; (2) misregistration of scenes; and (3) spectral similarity of ground cover. Error matrix probabilities were used to adjust the mapped area of classes, resulting in a decline of forest cover by 12.3% and increase of clearing by 18.5% (22.4 km(2) change; 95% confidence interval: 14.3-29.6 km(2)) between 1972 and 2004. Despite the mapping errors, Landsat data were able to identify broad patterns of land cover change that were verified from aerial photography. Most of the forest losses occurred in open forest to woodland habitat dominated by Eucalyptus, Corymbia, and Lophostemon species, which were largely replaced by sugarcane cropping. Melaleuca communities were similarly affected, though they have a much smaller distribution in the catchment. PMID- 18398637 TI - River-stream connectivity affects fish bioassessment performance. AB - Stream fish bioassessment methods assume that fish assemblages observed in sample sites reflect responses to local stressors, but fish assemblages are influenced by local factors as well as regional dispersal to and from connected streams. We hypothesized that fish movement to and from refugia and source populations in connected rivers (i.e., riverine dispersal) would weaken or decouple relations between fish community metrics and local environmental conditions. We compared fish-environment relations between streams that flow into large rivers (mainstem tributaries) and streams that lack riverine confluences (headwater tributaries) at multiple spatial grains using data from the USEPA's Environmental Monitoring and Assessment Program in the mid-Atlantic highlands, USA (n = 157 sites). Headwater and mainstem tributaries were not different in local environmental conditions, but showed important differences in fish metric responses to environmental quality gradients. Stream sites flowing into mainstem channels within 10 fluvial km showed consistently weaker relations to local environmental conditions than stream sites that lacked such mainstem connections. Moreover, these patterns diminished at longer distances from riverine confluences, consistent with the hypothesis of riverine dispersal. Our results suggest that (1) the precision of fish bioassessment metrics may be improved by calibrating scoring criteria based on the spatial position of sites within stream networks and (2) the spatial grain of fish bioassessment studies may be manipulated to suit objectives by including or excluding fishes exhibiting riverine dispersal. PMID- 18398638 TI - When coughing can cause stroke--a case-based update on cerebral air embolism complicating biopsy of the lung. AB - Introducing gas to the circulation is a largely iatrogenic problem which can result in serious morbidity and even death. We report a case of CT-guided needle biopsy of a pulmonary lesion complicated by acute stroke. The English literature on cerebral air embolism is reviewed, including an update of current opinions on its pathomechanism, diagnostic findings, therapeutic strategies, and means of prevention. PMID- 18398639 TI - The inflammatory response to femoral arterial closure devices: a randomized comparison among FemoStop, AngioSeal, and Perclose. AB - The objectives of this study were to investigate whether the systemic inflammatory response differs, in patients undergoing coronary angiography, among the arterial closure devices FemoStop, AngioSeal, and Perclose. The study is a prospective and randomized study. We measured pre- and postprocedural C-reactive protein (CRP), fibrinogen, and interleukin-6 (IL-6) plasma levels and collected clinical and procedural data on 77 patients who underwent coronary angiography because of stable angina pectoris. Patients were randomized to the following device: FemoStop (mechanical compression), AngioSeal (anchor and collagen sponge), or Perclose (nonabsorbable suture). No patient group experienced an increased incidence of vascular complications. There were no differences among the three groups regarding CRP, fibrinogen, or IL-6 values before or after coronary angiography. IL-6 levels increased 6 h after the procedure in all groups (p < 0.01), however, the increase did not differ among the groups. After 30 days there were no increased values of CRP or fibrinogen. We conclude that the femoral arterial closure devices AngioSeal and Perclose do not enhance an inflammatory response after a diagnostic coronary angiography, measured by CRP, fibrinogen, and IL-6, compared to femoral arterial closure using a mechanical compression device. PMID- 18398640 TI - Surgical research or comic opera? Let's give answers! PMID- 18398641 TI - Prospective randomized comparison of laparoscopic versus open adrenalectomy for sporadic pheochromocytoma. AB - BACKGROUND: Laparoscopic adrenalectomy for pheochromocytoma remains subject of debate, owing to the systemic consequences of pneumoperitoneum in patients with catecholamine-secreting tumors. METHODS: A prospective randomized study was conducted (2000-2006), evaluating cardiovascular instability during open (n = 9, group A) or laparoscopic (n = 13, group B) adrenalectomy for pheochromocytoma. Haemodynamic parameters were recorded by invasive monitoring. RESULTS: Haemodynamic instability was observed in 3/9 (group A) and 6/13 patients (group B), with a mean of 1.8 and 2.2 hypertensive peaks per patient (p = n.s.). Blood loss (164 +/- 94 cc versus 48 +/- 36 cc, p < 0.05) and operative time (180 +/- 40 versus 158 +/- 45 min, p = n.s.) favored laparoscopic procedures. Postoperative morbidity and mortality were nil. Hospital stay was shorter in group B (p < 0.05). Long-term follow-up was always normal. CONCLUSIONS: Laparoscopic approach for pheochromocytoma can be as safe as open surgery; intraoperative haemodynamic instability, although usually controlled with success, remains a source of concern. PMID- 18398642 TI - Quality control and educational value of laparoscopy-assisted gastrectomy in a high-volume center. AB - BACKGROUND: Laparoscopy-assisted gastrectomy (LAG) is increasingly performed in Japan by a number of surgeons. As this is a relatively new technique, a training system is important, however there has been little discussion about an educational system for teaching trainees to perform LAG and the ideal training system has not yet been established. PATIENTS AND METHODS: Two hundred and sixty three patients who underwent LAG at the Cancer Institute Hospital were included in this study. In all cases there was standardization of LAG (the way in which the surgical field was formed by the assistant and the way the operator dissected the lymph nodes was determined and all cases were performed using the same laparoscopic procedures) and a step-by-step training system was completed. Specialists performed the surgery in 213 patients (S group) while the remaining 50 patients had their surgery performed by trainees (T group). Early surgical outcomes were compared between specialists and trainees to clarify whether the standardization and our educational system are useful in maintaining the quality of LAG. RESULTS: T-group patients had significantly longer operation times than those of S-group (262.3 +/- 7.3 versus 233.3 +/- 3.7 min), however, the trainees reached the plateau of their learning curve earlier than previously reported. All other early surgical outcomes examined, including intraoperative blood loss (76.7 +/- 35.1 versus 64.9 +/- 7.7 ml), number of retrieved lymph nodes (33.4 +/- 1.4 versus 35.7 +/- 0.8), morbidity (8 versus 14%), and mortality (0% in both groups), were not significantly different between the two groups. CONCLUSION: The surgical results of T-group were almost equal to those of S-group, showing that our educational system is effective and surgical quality is maintained. Standardized laparoscopic procedures and sufficient intensive experience in the short term are requisites for effectively learning how to perform LAG. PMID- 18398643 TI - Randomized blinded trial comparing the cardiopulmonary effects of NOTES with standard laparoscopy in a porcine survival model. AB - BACKGROUND: Anecdotal reports of natural orifice translumenal endoscopic surgery (NOTES) procedures in patients are emerging. Whether the new procedure is truly less invasive is not known. We compared perioperative cardiopulmonary parameters during NOTES with standard laparoscopy. METHODS: Twelve swine were randomized to transgastric peritoneoscopy with air or diagnostic laparoscopy with CO(2). Cardiopulmonary parameters were invasively monitored. Animals were survived for 14 days. Data were analyzed by an investigator blinded to the procedure performed. Treatments were contrasted on the mean outcome using a repeated measures linear model. RESULTS: All experiments were successfully completed. No gastric leak or peritonitis resulted. Two hundred and fifty cubic centimeters of saline irrigation was adequate to decrease gastric contamination. Respiratory compromise requiring desufflation resulted in two laparoscopy and one endoscopy animal. Mean arterial oxygen saturation remained at baseline for the NOTES group and decreased by 1.5 +/- 1.5% in the laparoscopic group (p < 0.001). Mean arterial pH dropped significantly lower in the laparoscopy versus the NOTES group (-0.09 +/- 0.06 versus -0.05 +/- 0.05, p = 0.01). Mean systolic blood pressure (Sbp) in both groups remained unchanged from the respective baseline (p = 0.45). Mean diastolic blood pressure (Dbp) showed a mean difference of 19 +/- 1.5 mmHg between the groups (p < 0.001), increasing from baseline for the laparoscopy group and decreasing significantly from baseline in the NOTES group (+2.4 +/- 12.5 versus -6.1 +/- 7.0 mmHg, p < 0.001). Mean heart rate increased significantly from baseline in the laparoscopy group compared to the NOTES group (15.0 +/- 23.4 versus 3.8 +/- 19.0 bpm, p = 0.004). A widening pulse pressure resulted in the NOTES group compared to the laparoscopy group (p < 0.001). The mean bladder pressure was 14.6 +/- 8.0 cmH(2)O in the laparoscopy group compared to 7.1 +/- 7.1 cm H(2)O the NOTES group (p < 0.001). CONCLUSION: Significant differences in measured but not clinically apparent cardiopulmonary parameters were encountered. The difference in insufflation gas, different vagal or catecholamine response may be contributing to these findings. PMID- 18398644 TI - Robotic total mesorectal excision for rectal cancer: it may improve survival as well as quality of life. PMID- 18398645 TI - Scarless cholecystectomy with combined transvaginal and transumbilical approach in a series of 20 patients. AB - BACKGROUND: Laparoscopic surgery has dramatically improved surgical care of patients reducing postoperative pain, wound infection rate, hospital stay, inability to work, risk of hernia, and cosmetic result. Natural orifice transluminal endoscopic surgery (NOTES) is even less traumatic to the abdominal wall and might further improve minimal invasive surgery of patients, but might also increase surgical risk when used by a transgastric or transcolonic approach with flexible endoscopes. Therefore we decided to use a transvaginal approach using rigid laparoscopic instruments for cholecystectomies. METHODS: Through a 5 mm incision deep in the umbilicus a pneumoperitoneum was created. The optic and a dissector were inserted through the posterior fornix of the vagina under laparoscopic control from the umbilicus and a transvaginal gallbladder removal was performed. RESULTS: 20 patients were successfully operated in a 4.5-month period. Operating time was 62 (35-100) min. No intra- or postoperative complications occurred in any patient. Gynecological examination after 8 days showed no negative findings and the cosmetic result was ideal with no visible scars. CONCLUSION: In our series we showed that cholecystectomies can be routinely performed in a NOTES technique without visible scar. The transvaginal approach is the safe in NOTES and common laparoscopic instruments can be used as long as there are no better flexible endoscopes for this purpose. PMID- 18398646 TI - How reliable is intraoperative cholangiography as a method for detecting common bile duct stones? : A prospective population-based study on 1171 patients. AB - BACKGROUND: Although intraoperative cholangiography (IOC) is a widely used method for detecting common bile duct stones (CBDS), its accuracy has not been fully evaluated in large nonselected patient samples. The purpose of this study was to assess the sensitivity, specificity and predictive value of dynamic IOC regarding its ability to diagnose CBDS in a population-based setting, and to assess the morbidity associated with the investigation. METHODS: All patients operated on for gallstone disease between 2003 and 2005 in the county of Uppsala in Sweden, a county with a population of 302,000 in December 2004, were registered prospectively. The outcome of cholangiography was validated against the postoperative clinical course. RESULTS: 1171 patients were registered, and among these IOC was performed in 1117 patients (95%). Common bile duct stones were found in 134 patients (11%). One perforation of the common bile duct caused by the IOC catheter was recorded. Sensitivity was 97%, specificity 99%, negative predictive value 99%, positive predictive value 95%, and overall accuracy 99%. In 7 of the 134 cases where IOC indicated CBDS, no stones could be verified on exploration. In 4 of the 979 cases where IOC was normal, the clinical course indicated overlooked CBDS. CONCLUSION: Intraoperative cholangiography is a safe and accurate method for detecting common bile duct stones. PMID- 18398647 TI - Impact of laparoscopic surgery on bladder and sexual function after total mesorectal excision for rectal cancer. AB - BACKGROUND: Bladder and sexual dysfunction are well-documented complications of rectal cancer surgery. This study aimed to determine whether laparoscopy can improve the outcome of these dysfunctions or not. METHODS: The study included 63 of the 116 patients who underwent surgery for rectal cancer between 2002 and 2006. Bladder and male sexual function were studied by means of a questionnaire on the basis of the International Prostatic Symptom Score (IPSS) and International Index of Erectile Function (IIEF). In addition, bladder function was determined by means of postvoid residual urine measurement and uroflowmetry. Postoperative functions were compared with the preoperative data to detect subjective functional deterioration. Outcomes were compared between patients who underwent open (group 1, n = 29) and laparoscopic (group 2, n = 34) total mesorectal excision. RESULTS: Only minor disturbances of bladder function were reported for one patient (3%) in group 1 and three patients (9%) in group 2 (p > 0.05). Impotency after surgery was experienced by 6 of 17 preoperatively sexually active males (29%) in group 1 and 1 of 18 males (5%) in group 2 (p = 0.04). Similarly, 5 of 10 women (50 %) in group 1 and 1 of 14 women (7%) in group 2 felt that their overall level of sexual function had decreased as a result of surgery (p = 0.03). CONCLUSIONS: Open rectal cancer resection is associated with a higher rate of sexual dysfunction, but not bladder dysfunction, compared with laparoscopic surgery. Laparoscopic rectal cancer surgery offers a significant advantage with regard to preservation of postoperative sexual function and constitutes a true advance in rectal cancer surgery compared with the open technique. The proposed advantages can be attributed to improvement in visibility by the magnification feature of laparoscopic surgery. PMID- 18398649 TI - Endoscopic resection of ampullary tumors: 12-year review of 21 cases. AB - BACKGROUND: Endoscopic snare papillectomy is increasingly performed with curative intent for benign papillary tumors. This study aimed to evaluate the outcome of endoscopic resection for ampullary tumors at a single center. METHODS: All ampullary tumors without macroscopic features of malignancy identified by the endoscopic retrograde cholangiopancreatography (ERCP) from January 1995 to February 2007 were included in the study. Papillectomy was performed by snare resection using electrocautery. Argon plasma coagulation was effective for fulguration of small tissue remnants not amenable to snare resection. RESULTS: Of the 21 patients (9 men and 12 women; mean age, 67.2 +/- 14.3 years) evaluated, 11 had adenoma (7 had low-grade dysplasia [LGD] and 4 had high-grade dysplasia [HGD]), and 10 had carcinoma. All the patients underwent papillectomy. Of the 21 patients, 18 had extraductal growth or minimal intraductal growth, and 3 had extensive intraductal growth. The endoscopic complications (23.8%) included one case of mild bleeding, two cases of mild pancreatitis, and two cases of moderate pancreatitis. After papillectomy, 15 patients underwent Whipple procedures (endoscopic failure, 74.1%), including 3 patients with extensive intraductal growth (complete removal of the lesion impossible), 9 patients with carcinoma beyond the mucosal layer, and 3 patients with recurrence treated surgically. Endoscopic success (28.5%) was obtained for the remaining six patients (4 with LGD and 2 with HGD). Papillectomy was determined to be curative after a mean follow-up period of 15.9 +/- 14.9 months. CONCLUSIONS: In the hands of an experienced endoscopist, endoscopic papillectomy is a clinically effective treatment for ampullary tumors without invasive neoplasia. Evaluation of a prepapillectomy tumor extension is an important criterion for assessment of endoscopic success. PMID- 18398648 TI - A meta-analysis of ambulatory versus inpatient laparoscopic cholecystectomy. AB - BACKGROUND: Laparoscopic cholecystectomy is increasingly used on an ambulatory basis. This study aimed to examine its effectiveness for carefully selected patients. METHODS: A systematic review of Cochrane, Embase, and Medline using the keywords "ambulatory," "laparoscopic," and "cholecystectomy" was performed. Postoperative complications leading to admissions and readmissions were compared between day care and inpatient laparoscopic cholecystectomy groups. Postoperative quality of life, patient satisfaction, and cost effectiveness also were analyzed. RESULTS: The search process identified seven clinical trials suitable for meta analysis. These trials, consisting of 598 patients, compared day care and inpatient procedures. The unplanned admission rate in the ambulatory group was comparable with the prolonged hospitalization of inpatients (odds ratio [OR], 1.979; 95% confidence interval [CI], 0.846-4.628). There was no significant difference between the readmission rates of the two groups (OR, 0.964; 95% CI, 0.318-2.922). The quality-of-life indicators were similar for the ambulatory and overnight-stay patients (p = 0.195). The cost effectiveness was better for the day care procedures because of the shorter mean hospital stay. CONCLUSION: Ambulatory laparoscopic cholecystectomy can be performed safely for selected patients, with reduced cost and a high level of patient satisfaction. PMID- 18398650 TI - Sixty-month follow-up after endoscopic inguinal hernia repair with three types of mesh: a prospective randomized trial. AB - BACKGROUND: This prospective, clinical, randomized, double-blind study was intended to investigate the impact of the structure and the amount of polypropylene (PP) mesh used in laparoscopic transabdominal preperitoneal hernioplastic (TAPP) on physical function and life quality. METHODS: 180 male patients with primary inguinal hernia undergoing TAPP were randomized for using a heavyweight (108 g/m(2)), double-filament PP mesh (Prolene, 10 x 15 cm, group A, n = 60), a multifilament, heavyweight variant (116 g/m(2)) of PP mesh (Serapren, 10 x 15 cm, group B, n = 60), or a composite mesh (polyglactin and PP) (Vypro II, 10 x 15 cm, group C, n = 60). We compared in terms of complications (seromas, recurrence rate) and life quality (pain development, physical function). The development of life quality was documented according to the SF-36 Health Survey. The follow-up period was 60 months. RESULTS: The recurrence rate (2.2% overall) during 60-month follow-up was not significantly different between the groups. Convalescence in group A was slower than in groups B and C: mean-term values of the visual scales for pain development were significantly (p < 0.05) higher, incapacity for work was 8.2 days longer, and urological adverse effects were stronger. The mean-term development of life quality was significantly lower in group A up to 12th week postoperatively. There were no significant differences between groups B and C. Beyond the 12th postinterventional week the differences diminished. CONCLUSIONS: The composite mesh does not provide an advantage concerning physical function or pain development in comparison to the multifilament, heavyweight, pure polypropylene mesh. Independently of which mesh was implanted 5% of patients are still suffering from discomfort after 5 years. PMID- 18398651 TI - Influence of wrap length on the effectiveness of Nissen and Toupet fundoplication: a prospective randomized study. AB - BACKGROUND: Incontinence or hypercontinence of the fundic wrap depends primarily on the length of the valve or the type of procedure. Much less attention has been paid to the fundic wrap length. This study aimed to compare the effectiveness of two different wrap lengths among the patients undergoing partial or total fundoplication. METHODS: For this study, 153 patients were randomized to either Nissen (1.5- or 3-cm wrap) or Toupet (1.5- or 3-cm wrap) laparoscopic fundoplication. The groups were compared according to intensity of dysphagia, esophageal manometry data, ambulatory 24-h pH monitoring data, postoperative esophagitis rate, and overall treatment failure rate. RESULTS: In all the groups, the tone of the lower esophageal sphincter was significantly increased and the DeMeester score significantly decreased, reaching normal levels. At 6 months after surgery, the Toupet 1.5-cm group had significantly more cases of esophagitis than the 3-cm wrap group (24.2% vs 3.3%; p<0.05). At 12 months after surgery, only one patient in the Nissen 3-cm group had moderate to severe dysphagia. In all cases, failures were associated with persistent erosive esophagitis. At the 12-month follow-up assessment, treatment failures were significantly more common in Toupet 1.5-cm group than in the Toupet 3-cm group (17.5% vs 2.7%; p<0.05). However, such differences were not observed in the Nissen groups (7.8% for 1.5 cm and 15.6% for 3 cm; p>0.05). CONCLUSIONS: Evaluation of the treatment results suggests that the wrap length is important in partial Toupet fundoplication to avoid treatment failures. The 3-cm wrap is superior to the 1.5-cm wrap in cases of partial posterior Toupet fundoplication. The influence of wrap length on treatment failure remains unconfirmed for the Nissen procedure. PMID- 18398652 TI - Minimal access surgery of pediatric inguinal hernias: a review. AB - Inguinal hernia is a common problem among children, and herniotomy has been its standard of care. Laparoscopy, which gained a toehold initially in the management of pediatric inguinal hernia (PIH), has managed to steer world opinion against routine contralateral groin exploration by precise detection of contralateral patencies. Besides detection, its ability to repair simultaneously all forms of inguinal hernias (indirect, direct, combined, recurrent, and incarcerated) together with contralateral patencies has cemented its role as a viable alternative to conventional repair. Numerous minimally invasive techniques for addressing PIH have mushroomed in the past two decades. These techniques vary considerably in their approaches to the internal ring (intraperitoneal, extraperitoneal), use of ports (three, two, one), endoscopic instruments (two, one, or none), sutures (absorbable, nonabsorbable), and techniques of knotting (intracorporeal, extracorporeal). In addition to the surgeons' experience and the merits/limitations of individual techniques, it is the nature of the defect that should govern the choice of technique. The emerging techniques show a trend toward increasing use of extracorporeal knotting and diminishing use of working ports and endoscopic instruments. These favor wider adoption of minimal access surgery in addressing PIH by surgeons, irrespective of their laparoscopic skills and experience. Growing experience, wider adoption, decreasing complications, and increasing advantages favor emergence of minimal access surgery as the gold standard for the treatment of PIH in the future. This article comprehensively reviews the laparoscopic techniques of addressing PIH. PMID- 18398653 TI - Silent Cerebral Infarct Transfusion (SIT) trial imaging core: application of novel imaging information technology for rapid and central review of MRI of the brain. AB - The Silent Cerebral Infarct Multicenter Transfusion (SIT) Trial is a multi institutional intervention trial in which children with silent cerebral infarcts are randomized to receive either blood transfusion therapy or observation (standard care) for 36 months. The SIT Trial is scheduled to enroll approximately 1,880 children with sickle cell disease from 29 clinical sites in the United States, Canada, UK, and France. Each child undergoes a screening magnetic resonance imaging (MRI) of the brain to detect the presence of silent cerebral infarct-like lesions, a pre-randomization (baseline) MRI and exit MRI to determine if there are new or enlarged cerebral infarcts, using a designated, prospective imaging protocol. The objective of this manuscript is to describe the innovative method used to process and adjudicate imaging studies for an international trial with a primary endpoint that includes neuroimaging. Institution investigators at each site were provided with computer hardware and software for transmission of MRI images that allow them to strip the scans of all personal information and add unique study identifiers. Three neuroradiologists at separate academic centers review MRI studies and determine the presence or absence of silent cerebral infarct-like lesions. Their findings are subsequently placed on web-based case report forms and sent to the Statistical Coordinating Center. The average time from imaging center receipt of the MRI study to the radiology committee report back to the local site is less than two working days. This novel strategy was designed to maximize efficiency and minimize cost of a complex large multicenter trial that depends heavily on neuroimaging for entry criteria and assessment for the primary outcome measures. The technology, process, and expertise used in the SIT Trial can be adapted to virtually any clinical research trial with digital imaging requirements. PMID- 18398655 TI - Behavioral and olfactory responses of female Salaria pavo (Pisces: Blenniidae) to a putative multi-component male pheromone. AB - The peacock blenny, Salaria pavo (Risso 1810), typically breeds in rocky shores of the Mediterranean and adjacent Atlantic coast. Males defend a territory around a hole or cavity wherein females deposit eggs that the male guards until hatching. A pair of exocrine glands on the anal fin (anal glands) of males produces a putative pheromone involved in attraction of reproductively competent females to the nest. We used behavioral assays to assess species-specific attraction of reproductively competent females to putative male pheromones, including the anal gland pheromone. Additionally, chromatographic fractions of anal glands and male-conditioned water were tested for olfactory potency in females by electro-olfactogram analysis (EOG). In a flow-through tank or fluviarium, reproductive females were attracted to male-conditioned water and to the anal gland macerate from conspecifics but not to those of a closely related heterospecific. In addition, attraction of reproductive females to conspecific anal gland macerate occurred only during their initial upstream movement in the fluviarium; this was an ephemeral response when compared with the response to male-conditioned water that attracted females throughout the entire period of observation (5 min). Reproductive females also were attracted during the entire period of observation to water-conditioned by conspecific males whose anal glands had been removed. However, the attraction was more variable than that to water conditioned by intact males. Moreover, females were not attracted to male (without anal glands) odor during their initial upstream movement in the fluviarium. Finally, non-reproductive females were not attracted to the conspecific anal gland macerate. The EOG responses of females to molecular weight fractions and solid-phase extraction and high-performance liquid chromatography fractions of anal gland macerates and male-conditioned water (with and without anal glands) suggest that the anal glands release hydrophilic odorants that consist mainly of molecules smaller than 500 Da. Furthermore, males released potent odorants that do not originate from the anal glands. We hypothesize that females respond to a multi-component male pheromone to find mates. The putative anal gland pheromone is possibly comprised of hydrophilic odorants, whereas the other component(s), presumably of gonadal origin, may be less water-soluble. PMID- 18398656 TI - Attraction to herbivore-induced plant volatiles by the host-foraging parasitoid fly Exorista japonica. AB - Responses of the tachinid fly Exorista japonica Townsend to odors from corn plants infested with the fly's host, the larvae of the noctuid moth Mythimna separata (Walker), were examined in a wind tunnel. Naive female flies showed a higher rate of landing on M. separata-infested corn plants from which the host larvae had been removed than on artificially damaged or intact corn plants. When paper impregnated with a solution of headspace volatiles collected from host infested plants was attached to intact plants, females landed on the plants at a high rate. Females also responded to intact plants to which had been attached with paper impregnated with a synthetic blend of nine chemicals identified previously in host-infested plants. There was an optimum concentration of the synthetic blend for the females' landing. Of the nine chemicals identified previously, four [(E)-4,8-dimethyl-1,3,7-nonatriene, indole, 3-hydroxy-2 butanone, and 2-methyl-1-propanol] released only by host-infested plants were classified as a host-induced blend. The other five [(Z)-3-hexen-1-yl acetate, (E) 2-hexenal, hexanal, (Z)-3-hexen-1-ol, and linalool] were classified as a non specific blend released not only by infested plants but also by artificially damaged or intact plants. In the wind tunnel, E. japonica females did not respond to intact plants to which paper containing a solution of non-specific blend or host-induced blend was attached. However, they showed a high level of response to a mixture of the non-specific and host-induced blends. These results indicate that naive E. japonica use a combination of non-specific and host-induced blends as an olfactory cue for locating host-infested plants. PMID- 18398657 TI - A common misuse of stepwise regression in studies of ratio of metastatic lymph nodes for gastric cancer. PMID- 18398658 TI - Surgical treatment in the management of malignant pleural mesothelioma: a single institution's experience. AB - BACKGROUND: A minority of patients with malignant pleural mesothelioma (MPM) are considered for surgery. To achieve a microscopically radical resection, combination with other treatment modalities is mandatory. The most effective combination is unknown. In our institute we have retrospectively analyzed the results of two combined modality regimens containing surgery. PATIENTS: Between January 2002 and September 2005, 15 MPM patients were treated with extrapleural pneumonectomy (EPP) and postoperative hemithoracic radiation (RT; 54 Gy). Previously, between January 1999 and December 2001, 20 patients underwent a combination of cytoreductive surgery - pleurectomy or EPP - and intraoperative hyperthermic intrathoracic chemotherapy (HITHOC), followed by radiotherapy to the thoracotomy scar and drainage tracts (24 Gy). RESULTS: The median operating time for EPP/RT was shorter (5.3 versus 6.9 h; P < 0.0001). Postoperative complications occurred in 8 EPP/RT patients (53%) and in 14 HITHOC patients (70%). Two HITHOC patients died postoperatively. Median overall survival was 29 months for EPP/RT patients and 11 months for HITHOC patients (P = ns). The median time to local recurrence was not reached for EPP/RT patients, and was 9 months for HITHOC patients (P = 0.003). Local control was achieved in ten EPP/RT patients (67%) with a follow-up of 5-59 months compared to four HITHOC patients (20%) with a follow-up of 4-27 months. CONCLUSIONS: In highly selected patients local control can be achieved with combination therapy but is accompanied by a high rate of (surgical) complications. Distant failure rates warrant further studies exploring the role of systemic chemotherapy while the use of cytoreductive surgery with intraoperative chemoperfusion for MPM is not supported. PMID- 18398659 TI - Urban/rural residence moderates effect of race on receipt of surgery in patients with nonmetastatic breast cancer: a report from the South Carolina central cancer registry. AB - BACKGROUND: Surgical resection is the cornerstone of therapy in patients with nonmetastatic breast cancer. Previous studies have reported underuse of adjuvant therapy among African Americans (AA). This study explores the independent effect of race on surgical resection in a recent, population-based sample of breast cancer patients. METHODS: All cases of nonmetastatic breast cancer reported to the our state Cancer Registry between 1996 and 2002 were identified and linked to the state Inpatient/Outpatient Surgery Files and the 2000 Census. Characteristics between Caucasian and AA patients were compared using Student's t and chi-square tests. Odds ratios (OR) of resection and 95% confidence intervals (CI) were calculated using logistic regression. RESULTS: We identified 12,404 Caucasian and 3,411 AA women. AA patients were more likely to be younger, non-married, have greater comorbidity, reside in rural communities, be less educated, live in poverty, and be uninsured or covered by Medicaid (all P < 0.0001). AA patients were slightly less likely to undergo resection compared to Caucasian patients (94.9% versus 96.4%, P < 0.0001). An interaction effect between race and urban/rural patient residence was observed (P = 0.003). After controlling for other factors, the adjusted OR for resection for urban AA patients was 0.58 (95% CI 0.41-0.82). In contrast, race had no effect on resection among rural patients (OR = 1.02; 95% CI 0.70-1.47). CONCLUSIONS: AA race is an independent predictor of underuse of surgery among urban patients with breast cancer, while rural residence is associated with underuse of surgery, irrespective of race. Interventions designed to optimize surgical cancer care should target these vulnerable populations. PMID- 18398660 TI - Perioperative use of beta-blockers and COX-2 inhibitors may improve immune competence and reduce the risk of tumor metastasis. AB - BACKGROUND: COX inhibitors and beta-blockers were recently suggested to reduce cancer progression through inhibition of tumor proliferation and growth factor secretion, induction of tumor apoptosis, and prevention of cellular immune suppression during the critical perioperative period. Here we evaluated the perioperative impact of clinically applicable drugs from these categories in the context of surgery, studying natural killer (NK) cell activity and resistance to experimental metastases. METHODS: F344 rats were treated with COX-1 inhibitors (SC560), COX-2 inhibitors (indomethacin, etodolac, or celecoxib), a beta-blocker (propranolol), or a combination of a COX-2 inhibitor and a beta-blocker (etodolac and propranolol). Rats underwent laparotomy, and were inoculated intravenously with syngeneic MADB106 tumor cells for the assessment of lung tumor retention (LTR). Additionally, the impact of these drug regimens on postoperative levels of NK cytotoxicity was studied in peripheral blood and marginating-pulmonary leukocytes. RESULTS: Surgery increased MADB106 LTR. COX-2 inhibition, but not COX 1 inhibition, reduced postoperative LTR. Etodolac and propranolol both attenuated the deleterious impact of surgery, and their combined use abolished it. Surgery decreased NK cytotoxicity per NK cell in both immune compartments, and only the combination of etodolac and propranolol significantly attenuated these effects. Lastly, the initiation of drug treatment three days prior to surgery yielded the same beneficial effects as a single pre-operative administration, but, as discussed, prolonged treatment may be more advantageous clinically. CONCLUSIONS: Excess prostaglandin and catecholamine release contributes to postoperative immune-suppression. Treatment combining perioperative COX-2 inhibition and beta blockade is practical in operated cancer patients, and our study suggests potential immunological and clinical benefits. PMID- 18398661 TI - HER2-positive early breast cancer and trastuzumab: a surgeon's perspective. AB - Although the treatment of patients with early-stage breast cancer is provided by a multidisciplinary team, surgeons must ensure they are well informed about all aspects of patient care. For example, understanding the importance of human epidermal growth factor receptor 2 (HER2) gene amplification and/or protein overexpression and the effect on patient prognosis can guide therapeutic decision making. In addition, surgeons should also be knowledgeable about the wide variety of available postsurgical treatments, from traditional chemotherapy and radiotherapy to novel agents such as the HER2-targeted monoclonal antibody trastuzumab. Trastuzumab was recently approved for adjuvant treatment of invasive, HER2-positive, node-positive breast cancer. Its approval was based on the combined results of two large-scale trials, which demonstrated that adding trastuzumab to chemotherapy significantly improves disease-free and overall survival. Knowledge about the indications, schedules, and safety profiles of therapies such as trastuzumab will enable surgeons to optimize the timing of surgery in relation to these therapies, and to make informed decisions about the team member to whom a particular patient should ideally be referred for postsurgical care. In the future, results from large-scale trials evaluating the clinical utility of neoadjuvant trastuzumab will become available. Early results from ongoing phase III trials of the addition of trastuzumab to presurgical chemotherapy suggest that high response rates, including pathologic complete responses, are achievable. If trastuzumab is approved for use in neoadjuvant regimens, the need for surgeons to be well informed about the appropriate use of this particular agent will become even more important. PMID- 18398663 TI - Resectable well-differentiated versus dedifferentiated liposarcomas: two different diseases possibly requiring different treatment approaches. AB - BACKGROUND: Division of retroperitoneal liposarcoma (RPLS) into well differentiated (WD) and dedifferentiated (DD) subtypes is established; however, WD and DD are usually treated similarly. We hypothesized that WD and DD have distinct biological behaviors mandating different treatments. METHODS: A prospective sarcoma database identified all primary/recurrent RPLS treated between 1996 and 2007: 77 DD (52%) and 58 WD (39.2%) patients were analyzed for recurrence rate, recurrence free survival (RFS), and overall survival (OS). RESULTS: At presentation, WD were mostly primary whereas DD were mostly recurrent (75.9% versus 58.4%; p = 0.04). A significant proportion of DD (37.7%) received chemotherapy compared to WD (1.7%; p < 0.0001). Multivisceral resection was more common in DD versus WD (45.5% versus 31%; p = 0.01). Gross total resection rates were equivalent (WD: 86.2%; DD: 85.7%). Overall and local recurrence were higher in DD versus WD (82.2% versus 50% and 71.2% versus 46.3%; p < 0.0001). Only 3.7% WD recurred as high grade metastatic disease. Median time to recurrence was 55.5 months in WD versus 13.5 months in DD (p < 0.0001). RFS and OS (1, 2, and 5 year) were higher in WD than DD (80.3% versus 55.9%; 65.1% versus 34.1%; 41.9% versus 7.8%; p < 0.0001) and (98% versus 88.1%; 95.6% versus 71.9%; 92.1% versus 36.5%; p < 0.0001) respectively. CONCLUSION: WD and DD have distinct biological behaviors. Gross total resection is achievable in most WD; unlike DD, high-grade recurrence is uncommon. Treatment should therefore reflect these biologic differences by maximizing survivorship while avoiding unnecessarily extensive multivisceral resection. SYNOPSIS: The biological behaviors of well differentiated and dedifferentiated liposarcomas differ significantly. This article presents outcomes of two different surgical approaches that were implemented at the UTMDACC, treating these tumors as different disease entities. PMID- 18398664 TI - Computer-assisted provision of emergency contraception a randomized controlled trial. AB - BACKGROUND: Emergency contraception (EC) can prevent unintended pregnancy. However, many women continue to lack information needed to use EC effectively and clinician time to counsel women about EC is limited. OBJECTIVE: To evaluate whether computer-assisted provision of EC can increase knowledge and use of EC among women able to access EC without a prescription. DESIGN: We conducted a randomized controlled trial in which the intervention group received a 15-minute computerized educational session and 1 pack of EC. The control group received education about periconception folate supplementation, but no information about EC. Participants were contacted 7 months after enrollment. PARTICIPANTS: Four hundred forty-six women recruited from 2 urgent care clinics in San Francisco in 2005. MEASUREMENTS: Knowledge of EC, use of EC, and self-reported pregnancy. RESULTS: At follow-up, women in the intervention group answered an average of 2 more questions about EC correctly than they had at baseline, whereas women in the control group answered only 1 more item correctly (2.0 vs 1.2, p < .001). There was a trend toward more use of EC during the study period in the intervention group (10% vs 4% of women followed, p = .06; 6% vs 3%, p = .09 of women enrolled). Fewer women in the intervention group were pregnant at the time of follow-up (0.8% vs 6.5%, p = .01 of women followed; 0.5% vs 4.0%, p = .01 of women enrolled). CONCLUSIONS: Computer-assisted provision of EC in urgent care waiting areas increased knowledge of EC in a state where EC had been available without a prescription for 3 years. PMID- 18398665 TI - Nestin-positive spheres derived from canine bone marrow stromal cells generate cells with early neuronal and glial phenotypic characteristics. AB - Bone marrow stromal cells (BMSCs) isolated from humans and rodents have been shown to generate neural cells under specific culture conditions and after transplantation in the central nervous system. The apparent plasticity of BMSCs has therefore been a target of intensive research in attempt to develop a novel therapy for neurological diseases. Canines sustain neurological disorders (e.g., traumatic spinal cord injury) that closely mirror pathology of those in humans. Therefore, we evaluated neural differentiation properties of canine BMSCs to provide insights into basic characterization of these cells for future neurotransplantation trials in canine patients with neurological disorders. We demonstrate that canine BMSCs form spherical cellular aggregates on anti-adhesive culture substrate in serum-free culture media, which morphologically and phenotypically resemble spherical aggregates of neural progenitor cells, so called neurospheres. Upon dissociation and subculture on adhesive substrate, canine BMSCs express neuronal (ss capital SHA, Cyrillic-tubulin) and glial (GFAP, A2B5, and CNPase) markers. Formation of spherical aggregates appears to be a critical preceding process for some of the glial marker expression (CNPase and A2B5). However, expression of more mature neuronal (MAP2) and glial (MBP) markers could not be induced with the protocol used in this study. We suggest that induction of canine BMSCs into cells with neural progenitor cell characteristics is possible and that these cells may have the potential for future cellular therapy for neurological disorders. PMID- 18398666 TI - Rapid and quantitative detection of p38 kinase pathway in mouse blood monocyte. AB - The p38 alpha mitogen-activated protein kinase (MAPK) is essential in controlling the production of many proinflammatory cytokines, and its specific inhibitor can effectively block their production for treating human diseases. To effectively identify highly specific p38 alpha inhibitors in vivo, we developed an ex vivo mouse blood cell-based assay by flow cytometry to measure the intracellular p38 alpha kinase activation. We first attempted to identify the individual blood cell population in which the p38 alpha kinase pathway is highly expressed and activated. Based on CD11b, combined with Ly-6G cell surface expression, we identified two distinct subsets of non-neutrophilic myeloid cells, CD11b(Med)Ly 6G(-) and CD11b(Lo)Ly-6G(-), and characterized them as monocytes and natural killer (NK) cells, respectively. Then, we demonstrated that only monocytes, not NK cells, expressed a high level of p38 alpha kinase, which was rapidly activated by anisomycin stimulation as evidenced by the phosphorylation of both p38 and its substrate, MAPKAP-K2 (MK2). Finally, the p38 alpha kinase pathway activation in monocytes was fully inhibited by a highly selective p38 alpha kinase inhibitor dose-dependently in vitro and in vivo. In conclusion, we demonstrated an effective method for separating blood monocytes from other cells and for detecting the expression level and activation of the p38 alpha kinase pathway in monocytes, which provided a new approach for the rapid identification of specific p38 alpha inhibitors. PMID- 18398667 TI - Has laparoscopic bariatric surgery been accepted in Japan? The experience of a single surgeon. AB - BACKGROUND: Obesity is steadily increasing in Asia due to factors such as a lack of exercise, adoption of a more Western diet, changing lifestyles, environments, or stresses. Even in Japan, this tendency is notable, and metabolic syndrome has become widely recognized. However, bariatric surgery is still uncommon in Japan. There are no adequate data regarding the experience and outcome of bariatric surgery in Asia. Here, we report on the current status of morbid obesity and the outcomes of bariatric surgery by a single surgeon in Japan. METHODS: Between February 2002 and January 2008, we have performed laparoscopic bariatric surgery for morbid obesity in 178 cases. They consisted of laparoscopic Roux-en-Y gastric bypass (LRYGBP) in 105 cases, laparoscopic sleeve gastrectomy (LSG) in 26 cases, laparoscopic sleeve gastrectomy with duodenal jejunal bypass (LSG/DJB) in 14 cases, laparoscopic adjustable gastric banding (LAGB) in 13 cases, and laparoscopic biliopancreatic diversion with duodenal switch in one case under the same protocol of follow up. The first author of this paper performed all procedures. RESULTS: One hundred and thirty-eight patients with a follow-up of over 3 months after surgery were enrolled. LRYGBP accounted for 72% of all bariatric procedures. The reduction of weight and body mass index (BMI) in LRYGBP and LSG showed similar results. These outcomes were superior to those of LAGB. Percentage of excess BMI loss (%EBMIL) of LRYGBP showed greater reductions at follow-ups 6, 9, 12, and 18 months after surgery compared to that of LRYGBP and LAGB. All procedures resulted in over 50% of %EBMIL after 18 months of follow-up. There was no postoperative mortality within 30 days after surgery. Preoperative comorbidity including diabetes mellitus, hypertension, and hyperlipidemia were resolved or improved after surgery in most patients. CONCLUSION: In bariatric surgery, LRYGBP is the most effective treatment for morbid obesity, while LAGB has a low risk of postoperative complications. LSG is also a safe procedure for supermorbidly obese patients. We expect that bariatric surgery will be a common procedure for patients with morbid obesity in Japan. PMID- 18398668 TI - Gastric cancer after laparoscopic adjustable gastric banding. AB - A 63-year-old woman with BMI 46 underwent laparoscopic gastric banding. In the standardized follow-up after 14 and 24 months, the GI series and gastroscopy showed no pathological signs. The patient had hematemesis 32 months after gastric banding, followed by symptoms of obstruction, for which a laparotomy was performed. At operation, peritoneal carcinomatosis due to gastric cancer was found. Symptoms after bariatric procedures can be similar to symptoms of gastric or esophageal cancer. We believe that yearly postoperative gastroscopy is indicated to exclude pathological changes. PMID- 18398670 TI - Synergistic effect between apolipoprotein E and apolipoprotein A1 gene polymorphisms in the risk for coronary artery disease. AB - Alterations in lipid metabolism and genetic predisposition are major risk factors for coronary artery disease (CAD). Variations in genes involved in lipid metabolism may act synergistically to confer risk or protection against CAD. The objective of the present study was to determine such interactions in variants of apolipoprotein E and apolipoprotein A1 genes. One hundred and forty subjects with clinically confirmed CAD and 100 unrelated normal subjects participated in the study. Multiple regression analysis was used to relate lipid and apolipoprotein profiles with genotypes. Odd ratios were calculated for various combinations of ApoE and ApoA1 genotypes. Prevalence of ApoE 'E4' and ApoA1 'A' and 'T' alleles was significantly higher in patients than controls. Serum apolipoprotein E and apolipoprotein A1 levels were significantly lower in CAD patients than controls. When lipid parameters were related to genotypes, the polymorphisms associated to various markers were in agreement with previous reports. ApoE 2/4 genotype in combination with either ApoA1 heterozygous GA or CT genotype conferred higher risk of CAD. E3 allele in homozygous or heterozygous state in combination with ApoA1+83 CC genotype conferred highest protection (P < 0.05). Thus, it appears that ApoE and ApoA1 gene variants may act synergistically to associate with risk and protection against CAD. PMID- 18398669 TI - Lipopolysaccharide induces cellular hypertrophy through calcineurin/NFAT-3 signaling pathway in H9c2 myocardiac cells. AB - Evidences suggest that lipopolysaccharide (LPS) participates in the inflammatory response in the cardiovascular system; however, it is unknown if LPS is sufficient to cause the cardiac hypertrophy. In the present study, we treated H9c2 myocardiac cells with LPS to explore whether LPS causes cardiac hypertrophy, and to identify the precise molecular and cellular mechanisms behind hypertrophic responses. Here we show that LPS challenge induces pathological hypertrophic responses such as the increase in cell size, the reorganization of actin filaments, and the upregulation of hypertrophy markers including atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) in H9c2 cells. LPS treatment significantly promotes the activation of GATA-4 and the nuclear translocation of NFAT-3, which act as transcription factors mediating the development of cardiac hypertrophy. After administration of inhibitors including U0126 (ERK1/2 inhibitor), SB203580 (p38 MAPK inhibitor), SP600125 (JNK1/2 inhibitor), CsA (calcineurin inhibitor), FK506 (calcineurin inhibitor), and QNZ (NFkappaB inhibitor), LPS-induced hypertrophic characteristic features, such as increases in cell size, actin fibers, and levels of ANP and BNP, and the nuclear localization of NFAT-3 are markedly inhibited only by calcineurin inhibitors, CsA and FK506. Collectively, these results suggest that LPS leads to myocardiac hypertrophy through calcineurin/NFAT-3 signaling pathway in H9c2 cells. Our findings further provide a link between the LPS-induced inflammatory response and the calcineurin/NFAT-3 signaling pathway that mediates the development of cardiac hypertrophy. PMID- 18398671 TI - Anti-proliferative and chemosensitizing effects of luteolin on human gastric cancer AGS cell line. AB - To investigate the anti-proliferative and chemosensitizing effects of luteolin on human gastric cancer, gastric cancer AGS cells were treated with luteolin and/or other chemotherapeutic agents. Cell growth was assessed by MTT assay, cell cycle and apoptosis were assessed by flow-cytometric analysis, and the expression of major proteins regulating cell cycle and apoptosis was also detected. The results showed that luteolin inhibited the growth of gastric cancer cells in a dose- and time-dependent manner. Flow cytometry revealed that the percentage of cells at G2/M phase increased dose-dependently. The protein levels of Cdc2, Cyclin B1 and Cdc25C were reduced and p21/cip1 was up-regulated after the treatment with luteolin. Furthermore, luteolin induced apoptosis in gastric cancer AGS cells. Western blotting showed that luteolin treatment significantly increased the levels of pro-apoptotic proteins, including Caspase-3, 6, 9, Bax, and p53, and decreased the levels of anti-apoptotic protein Bcl-2, thus shifting the Bax/Bcl ratio in favor of apoptosis. It was also demonstrated that a combinational treatment of cisplatin and luteolin induced more effectively cell growth inhibition, compared to cisplatin treatment alone. These findings indicate the anti-proliferative and chemosensitizing effects of luteolin on human gastric cancer AGS cells and luteolin may be a promising candidate agent used in the treatment of gastric cancer. PMID- 18398672 TI - Telomerase reconstitution contributes to resetting of circadian rhythm in fibroblasts. AB - The synchronization of the circadian signals to external or suprachiasmatic nucleus stimulation in the peripheral clocks is essential for maintaining the usual function of human body. However, aging will disrupt the synchronization of peripheral circadian rhythms, thus leading to some age-associated diseases. Up to now, little is known about the modification of the oscillatory rhythms in aged cells. A recent report showed that cell senescence in vascular human smooth muscle cells (HSMCs) altered circadian rhythms by a dysregulation of rhythmic gene expression. Furthermore, this alteration could be reversed by telomerase reconstitution. To test whether telomerase reconstitution can restore disrupted circadian rhythm in other types of senescent cells, we used fibroblasts as cell models to profoundly investigate the relationship between cell senescence and circadian rhythm modulation. We found that the response of rhythmic gene expression to serum stimulation was markedly attenuated in senescent fibroblasts, telomerase-reconstituted fibroblasts reset the circadian oscillation of rhythmic gene expression, and the activation of pERK-CREB and p38-CREB pathways might be involved in the circadian rhythm resetting. These findings suggested that telomerase reconstitution might be a good way to reset synchronization of peripheral circadian rhythms disrupted in senescent tissues. PMID- 18398674 TI - Spermine and endothelial damage during endotoxemia. AB - Diazenolate-NO-donors like spermine-NONOate use spermine as a nitric oxide releasing carrier. As one of the naturally occurring polyamines, spermine belongs to the most potent platelet aggregation inhibitors among the polyamines. Recent publications describe a dominating role for platelets and nitric oxide in the setting of endothelial dysfunction during endotoxemia. It is unknown whether spermine as single agent, beside his function as nitric oxide releasing carrier, exerts effects on macromolecular leakage during early endotoxemia. In male Wistar rats, macromolecular efflux was determined in mesenteric postcapillary venules using intravital microscopy at baseline, 60 and 120 min after the start of the experiment. In the first part of the experiments we investigated the effects of spermine and spermine-NONOate during leukocyte-independent states of endotoxemia, using the L- and P-selectin inhibitor fucoidin to suppress leukocyte-endothelial interaction. In the second part we investigated the effects of spermine on endothelial damage during endotoxemia without co-administration of fucoidin. Combined treatment of animals with spermine and fucoidin was as effective as treatment with spermine-NONOate and fucoidin alone. Spermine and fucoidin as single agents had no effects on macromolecular efflux during endotoxin challenge. Combined treatment of animals with spermine and fucoidin reduce macromolecular efflux to the same degree like treatment with spermine-NONOate and fucoidin. A currently unknown interaction between fucoidin and spermine leads to a significant reduction in macromolecular efflux. PMID- 18398673 TI - Amphiregulin: role in mammary gland development and breast cancer. AB - Extensive epithelial cell proliferation underlies the ductal morphogenesis of puberty that generates the mammary tree that will eventually fill the fat pad. This estrogen-dependent process is believed to be essentially dependent on locally produced growth factors that act in a paracrine fashion. EGF-like growth factor ligands, acting through EGF receptors are some of the principal promoters of pubertal ductal morphogenesis. Amphiregulin is the most abundant EGF-like growth factor in the pubertal mammary gland. Its gene is transcriptionally regulated by ERalpha, and recent evidence identifies it as a key mediator of the estrogen-driven epithelial cell proliferation of puberty: The pubertal deficiency in mammary gland ductal morphogenesis in ERalpha, amphiregulin, and EGFR knockout mice phenocopy each other. As a prognostic indicator in human breast cancer, amphiregulin indicates an outcome identical to that predicted by ERalpha presence. Despite this, a range of studies both on preneoplastic human breast tissue and on cell culture based models of breast cancer, suggest a possibly significant role for amphiregulin in driving human breast cancer progression. Here we summarise our current understanding of amphiregulin's contribution to mammary gland development and breast cancer progression. PMID- 18398675 TI - Stressful life events and depressive symptoms in a post-war context: which informal support makes a difference? AB - Gerontological literature utilizes the life stress paradigm to understand the impact of stress on psychological well-being, as well as the protective role that social resources play in buffering those effects; however these relationships are not well understood within various historical and social contexts. Utilizing a sample of 490 community-residing older adults in post-civil war Lebanon, this study investigates the moderating role of various social support factors in the stress-depression relationship. Contrary to expectations, results suggest that older Lebanese are more susceptible to the effects of health-decline and serious accident events than other types of stressors such as losses in the family and financial problems. Furthermore, findings provide evidence for a differential protective role for the respondent's spouse and children for only certain stressful events. The discussion highlights the role of family as a stress buffer in a shifting physical, social and political environmental context. Results from this study add to the discourse by emphasizing the importance of understanding the saliency of the stressor as well as source of support provided. PMID- 18398676 TI - The role of depression and anxiety symptoms in hospital readmissions after cardiac surgery. AB - The objective of this study was to determine the association between depression, anxiety and general stress symptoms with hospital readmissions after coronary artery bypass graft surgery. Two hundred and twenty six coronary artery bypass graft patients completed baseline self-report measures of depression, anxiety and stress and 222 patients completed these measures after surgery on the hospital ward. The hospital readmission outcomes at six months were analyzed using multivariable proportional hazard models. When analyzed as continuous variables in multivariable analyses, preoperative anxiety and postoperative depression predicted readmissions independent of medical covariates. In multivariable analyses with dichotomized anxiety, depression and stress, more than two-fold increase in readmission risk was attributable to preoperative anxiety and postoperative depression, independent of covariates. These results lend further support to previous research that has shown the symptoms of depression and anxiety are associated with morbidity following coronary artery bypass graft surgery. The findings highlight the need to develop suitable interventions for anxiety and depression among coronary artery bypass graft surgery patients. PMID- 18398677 TI - Toxicokinetic approach for the assessment of endocrine disruption effects of contaminated dredged material using female Carcinus maenas. AB - Ecotoxicological effects associated with contaminants present in dredged material from three Spanish ports, Ria of Huelva (SW, Spain), La Coruna (NW, Spain) and Bay of Cadiz (SW, Spain) were determined using a marine biotest based on endocrine disruption effects (vitellogenesis process alteration). Intermoult female Carcinus maenas were exposed in the laboratory to sediments from the Spanish ports per replicate during 21 days. Crab haemolymph samples were taken for vitellogenin/vitellin (VTG) analysis on days 0, 7, 14 and 21. Furthermore, chemical analysis was performed in the stations to determine the degree and nature of sediment contamination (Cr, Ni, Cu, Zn, Cd, Pb, Hg, As, PAHs and PCBs). A significant induction (P < 0.05) in vitellogenin/vitellin concentration was observed over time in individuals exposed to sediment samples containing significant concentrations of PAHs, PCBs, Hg, Pb and Zn. The toxicokinetic approach, proposed in this study related to the use of this biomarker in C. maenas, proved to be a powerful and sensitive tool to evaluate toxicity effects associated with contaminants present in dredged material. Moreover the integration of the results obtained through multivariate analysis approach (MAA) allowed the identification contaminants bound to sediments associated with adverse effects, validating the use of this marine biotest in a regulatory framework. PMID- 18398678 TI - Partial least squares regression: a valuable method for modeling molecular behavior in hemodialysis. AB - The aim of this work was to use the Partial Least Squares Regression (PLS) technique to fit simple models for the interpretation of an underlying complex process. In this study, the technique was used to build a statistical model for molecular kinetic data obtained from hemodialyzed patients. By using PLS we derived statistical linear models for the prediction of the equilibrated urea concentration which would be reached 30-60 min after the end of the dialysis session. Models with an average relative prediction error (RPE) of less than 0.05% were achieved. The model predictive accuracy was evaluated in a cross center study yielding an RPE < 3%. The chosen model was robust to variations such as sampling extraction time demonstrating a high capacity for modeling kinetics. It also was found to be useful for bedside monitoring. Finally, the PLS technique allowed identification of the most important co-variables in the model and of those patients with outlier patterns in their molecular dynamics. PMID- 18398679 TI - An integer programming model for optimizing shoulder rehabilitation. AB - Strength restoration is one goal of shoulder rehabilitation following rotator cuff repair surgery. However, the time spent in a physical rehabilitation setting is limited. The objective of this study was to develop a novel mathematical formulation for determining the optimal shoulder rehabilitation exercise protocol to restore normal shoulder strength given a time-constrained rehabilitation session. Strength gain was modeled using a linear dose-response function and biomechanical parameters of the shoulder musculature. Two different objective functions were tested: (1) one based on a least squares support vector machine using healthy and pathologic shoulder strengths (normative objective function), and (2) one which seeks to match the strength of the contralateral shoulder (contralateral objective function). The normative objective function was subject independent and the optimal protocol consisted of four sets each of adduction and external rotation. The contralateral objective function was subject-specific and the typical optimal protocol consisted of various set combinations of abduction and internal and external rotation. These results are only partially consistent with current practice. Improvement of the current model is dependent on a better understanding of strength training adaptation and shoulder rehabilitation. PMID- 18398680 TI - The effects of anisotropy on the stress analyses of patient-specific abdominal aortic aneurysms. AB - The local dilation of the infrarenal abdominal aorta, termed an abdominal aortic aneurysm (AAA), is often times asymptomatic and may eventually result in rupture an event associated with a significant mortality rate. The estimation of in-vivo stresses within AAAs has been proposed as a useful tool to predict the likelihood of rupture. For the current work, a previously-derived anisotropic relation for the AAA wall was implemented into patient-specific finite element simulations of AAA. There were 35 AAAs simulated in the current work which were broken up into three groups: elective repairs (n = 21), non-ruptured repairs (n = 5), and ruptured repairs (n = 9). Peak stresses and strains were compared using the anisotropic and isotropic constitutive relations. There were significant increases in peak stress when using the anisotropic relationship (p < 0.001), even in the absence of the ILT (p = 0.014). Ruptured AAAs resulted in elevated peak stresses as compared to non-ruptured AAAs when using both the isotropic and anisotropic simulations, however these comparisons did not reach significance (p(ani) = 0.55, p(iso) = 0.73). While neither the isotropic or anisotropic simulations were able to significantly discriminate ruptured vs. non-ruptured AAAs, the lower p-value when using the anisotropic model suggests including it into patient-specific AAAs may help better identify AAAs at high risk. PMID- 18398682 TI - A comparative analysis of coronary and aortic flow waveforms. AB - Empirical Mode decomposition (EMD) is a mathematical tool designed to analyze non stationary, non-linear stochastic waves. EMD separates a waveform into its constituent modes of oscillations or intrinsic mode functions (IMFs) and provides meaningful definitions of instantaneous frequency, instantaneous energy, mean trends and oscillation about the mean trends. This study provides a detailed mathematical analysis of blood flow waveforms in the porcine left anterior descending artery and aorta using EMD. Flow data with non-stationary and non linear characteristics were obtained for several hours using an implanted wireless biotelemetry device. EMD was validated against modern numerical techniques of principal component analysis (PCA) and wavelet analysis by comparing their predicted mean trends and energy distribution. EMD has an advantage over both techniques since it combines the strengths of both: it is adaptive (similar to PCA), and it can define instantaneous frequencies (similar to wavelet analysis). Because of the iterative nature, however, calculations using EMD can be computationally intensive. Sampling rate reduction was used to reduce computation time, without significantly effecting accuracy of IMF calculations. It was found that IMFs calculated at a sampling rate as low as 20 Hz were not significantly different (<6%) from those obtained at the original sampling rate (200 Hz). Our findings suggest that EMD may be a powerful mathematical tool to characterize flow waveforms. PMID- 18398681 TI - The differential effect of endothelial cell factors on in vitro motility of malignant and non-malignant cells. AB - Motility of cancer cells plays a critical role in tumor metastasis, and as such is a target for intervention. The motility of malignant Calu-1 human lung epithelial carcinoma cells is upregulated when placed on a human umbilical vein endothelial cell monolayer, while that of non-malignant L132 human lung epithelial cells is not. To dissect the factor(s) causing such differential behaviors, the motile responses of both cell lines to endothelial cell factors secreted to the media, on the endothelial cell surface, and secreted to the extracellular matrix-and to individual extracellular matrix proteins were compared. Cell motility was quantified by tracking the cell movement on a surface with time-lapse video microscopy, which was analyzed with the persistent random walk model of motility. None of the factors tested had a remarkable effect on L132 cell motility, but the Calu-1 cell motility was significantly upregulated by endothelial cell extracellular matrix and by laminin, fibronectin, collagen I and collagen VI individually. Flow cytometry analysis revealed significantly higher expression levels of integrin subunits beta1, alpha2, alpha3, and alpha6, which are known receptors for these extracellular matrix proteins, on the Calu-1 than L132 cells, implicating a role of these integrins in the observed motile behaviors of these cell lines. PMID- 18398683 TI - Integrin dependence of Calu-1 cell motility on endothelial extracellular matrix proteins. AB - We recently showed that the motility of the malignant Calu-1 human epidermoid lung carcinoma cells correlates to their expression levels of alpha2, alpha3, alpha6, and beta1 integrin subunits. To determine a causative relationship underlying this correlation, here we measured Calu-1 cell adhesion to and migration on laminin, collagen IV, human umbilical vein endothelial cell monolayers, and endothelial cell extracellular matrix in the presence of function blocking antibodies against the suspect integrin subunits. Blocking individual alpha subunits did not affect adhesion to or motility on laminin, but when used in pair-wise combinations, monoclonal antibody treatments significantly decreased tumor cell motility on, without diminishing adhesion to, laminin and the other substrates. Blocking all three alpha subunits at once or the beta1 subunit alone abolished migration on laminin; however, the latter treatment also abolished adhesion, whereas the former treatment did not. By contrast, blocking the beta1 subunit significantly reduced motility on collagen IV, endothelial cell monolayers, and endothelial cell extracellular matrix, but always without affecting adhesion. These results suggest a separation of roles and mechanisms of different integrins in adhesion and motility. PMID- 18398685 TI - Internal pH indicators for biomolecular NMR. AB - We describe a non-invasive technique for determining pH in biomolecular NMR sample using buffer components (formate, tris, piperazine, and imidazole) as internal pH indicators, whose (1)H NMR chemical shifts are sensitive to pH in a range from 2.5 to 9.8. This method is suitable for a wide range of applications where samples are handled intensively during NMR titrations or in high throughput analysis in structural genomics or metabolomics. PMID- 18398684 TI - Imaging synaptic inhibition in transgenic mice expressing the chloride indicator, Clomeleon. AB - We describe here a molecular genetic approach for imaging synaptic inhibition. The thy-1 promoter was used to express high levels of Clomeleon, a ratiometric fluorescent indicator for chloride ions, in discrete populations of neurons in the brains of transgenic mice. Clomeleon was functional after chronic expression and provided non-invasive readouts of intracellular chloride concentration ([Cl( )](i)) in brain slices, allowing us to quantify age-dependent declines in resting [Cl(-)](i) during neuronal development. Activation of hippocampal interneurons caused [Cl(-)](i) to rise transiently in individual postsynaptic pyramidal neurons. [Cl(-)](i) increased in direct proportion to the amount of inhibitory transmission, with peak changes as large as 4 mM. Integrating responses over populations of pyramidal neurons allowed sensitive detection of synaptic inhibition. Thus, Clomeleon imaging permits non-invasive, spatiotemporally resolved recordings of [Cl(-)](i) in a large variety of neurons, opening up new opportunities for imaging synaptic inhibition and other forms of chloride signaling. PMID- 18398686 TI - Arabidopsis thaliana mutants reveal a role for CSP41a and CSP41b, two ribosome associated endonucleases, in chloroplast ribosomal RNA metabolism. AB - A proteomic analysis of Chlamydomonas reinhardtii 70S ribosomes identified two proteins, RAP38 and RAP41, which associate in stoichiometric amounts with intact ribosomes. In this work we show results that suggest the Arabidopsis thaliana homologs, CSP41b and CSP41a, participate in ribosomal RNA metabolism. Csp41a-1 and csp41b-1 single mutants show little phenotype, while the loss of both proteins is lethal. Plants homozygous for the csp41b-1 mutation and heterozygous for the csp41a-1 mutation (csp41b-1/csp41a-1*) fail to accumulate CSP41b and show a marked reduction in the levels of CSP41a. These mutants have reduced chlorophyll content, grow slower and over-accumulate 23S precursor rRNAs compared to their wild-type (WT) siblings, whereas other rRNAs or mRNAs are unaffected. Chloroplast polysome assembly is reduced in csp41b-1/csp41a-1* mutants, which also contain increased amounts of pre-ribosomal particles compared to mature 70S ribosomes. Our results also indicate that CSP41b associates with pre-ribosomal particles in vivo. In vitro, the pattern of 23S precursors and mature rRNAs is altered upon incubation with recombinant CSP41a and CSP41b. Taken together, these results suggest that CSP41a and CSP41b have a role in chloroplast ribosomal RNA metabolism, most likely acting in the final steps of 23S rRNA maturation. PMID- 18398688 TI - Steroidogenic acute regulatory (StAR) protein and cholesterol side-chain cleavage (P450scc)-regulated steroidogenesis as an organ-specific molecular and cellular target for endocrine disrupting chemicals in fish. AB - Biologically active steroids are synthesised de novo in specialised cells of several organs, including the adrenal gland, testis, ovary, brain, placenta and adipose tissue. Regardless of organ or tissue, the rate-limiting step in steroid hormone synthesis is the movement of cholesterol across the mitochondrial membrane (i.e. from the outer to the inner membrane) mediated by the steroidogenic acute regulatory (StAR) protein. Subsequent conversion of cholesterol to pregnenolone by cytochrome P450 side-chain cleavage (P450scc) represents the initiation of steroidogenesis. Chemically mediated disruption of StAR and P450scc expression may represent the first step in the sequence of related event cascades underlying xenoestrogen-induced toxicity and transmittable disturbances to the whole organism level. This may include, but is not limited to, alterations in sexual differentiation, growth, reproduction, development and metabolism. Despite the integral role of StAR and P450scc in acute steroidogenesis, and popular demand from regulatory agencies, bioassays for evaluating the effect of endocrine-disrupting chemicals have the potential to overlook chemicals that may modulate estrogenic responses through mechanisms that do not involve direct binding to estrogen receptors (ERs). In addition to their effect as direct ER agonists, the effects of endocrine disruptors may be evaluated and interpreted as interference with steroidogenesis and with the steroidal regulation of the normal development and function of juvenile, male and female individuals. Knowledge of these effects is scarce, indicating that relatively little is known about the mechanisms or mode-of-action of chemical alterations to steroidogenesis and their potential toxicity for wildlife species. In addition, analytical methods for the complete adaptation of these responses as biomarkers of response and effect are yet to be properly validated. PMID- 18398687 TI - Phenotypic plasticity of neoplastic ovarian epithelium: unique cadherin profiles in tumor progression. AB - The mesodermally derived normal ovarian surface epithelium (OSE) displays both epithelial and mesenchymal characteristics and exhibits remarkable phenotypic plasticity during post-ovulatory repair. The majority of epithelial ovarian carcinomas (EOC) are derived from the OSE and represent the most lethal of all gynecological malignancies, as most patients (approximately 70%) present at diagnosis with disseminated intra-abdominal metastasis. The predominant pattern of EOC metastasis involves pelvic dissemination rather than lymphatic or hematologic spread, distinguishing EOC from other solid tumors. Acquisition of the metastatic phenotype involves a complex series of interrelated cellular events leading to dissociation (shedding) and dispersal of malignant cells. A key event in this process is disruption of cell-cell contacts via modulation of intercellular junctional components. In contrast to most carcinomas that downregulate E-cadherin expression during tumor progression, a unique feature of primary well-differentiated ovarian cancers is a gain of epithelial features, characterized by an increase in expression of E-cadherin. Subsequent reacquisition of mesenchymal features is observed in more advanced tumors with concomitant loss of E-cadherin expression and/or function during progression to metastasis. The functional consequences of this remarkable phenotypic plasticity are not fully understood, but may play a role in modulation of cell survival in suspension (ascites), chemoresistance, and intraperitoneal anchoring of metastatic lesions. PMID- 18398689 TI - Identification of air pollutant sampling period using horizontal dilution potential. AB - A new model is proposed for estimating horizontal dilution potential of an area using wind data. The mean wind speed and wind direction variation are used as a measure of linear and angular spread of pollutant in the atmosphere. The methodology is applied to monitored hourly wind data for each month of 1 year for wind data collected at Vadodara, Gujarat and monthly dilution potential is estimated. It is found that there is a gradual variation of horizontal dilution potential over a year with limited dilution during post monsoon period i.e., October and November and a high dilution in pre monsoon period i.e., May and June. This information can be used to design air quality sampling network and duration of sampling for source apportionment study. Air pollutant sampling during high dilution period can be carried out for identifying urban and rural dust and wind blown dust from mining activity. Air pollutant sampling during low dilution period can be carried out for capturing large amount of particulate matter from anthropogenic sources like elevated stack of furnace. PMID- 18398690 TI - Post-classification comparison of land cover using multitemporal Landsat and ASTER imagery: the case of Kahramanmaras, Turkey. AB - This study assessed land cover (LC) changes in Kahramanmaras (K.Maras) and its environs by using multitemporal Landsat and ASTER imagery, respectively belong to 1989, 2000 and 2004. A priori defined nine land cover classes in the classification scheme were urban and built-up, forest, sparsely vegetated areas, grassland, vegetated stream beds, unvegetated stream beds, bare areas, crop fields, and water bodies. Individual classifications were employed using the combination of both unsupervised and supervised classification methods. Iterative Self Organizing Data Analysis (ISODATA) was used to reduce spectral variation in the scenes arising from complex pattern of crop fields. Maximum Likelihood classifier was used in the LC classification of the individual images. Image pairs of consecutive dates were compared by overlaying the thematic LC maps and cross-tabulating the LC statistics. Urbanization and expansion of agriculture were the major reasons for the dramatic LC conversions. The amount of conversion from crop fields to water occurred as large as 927.67 ha, accounting for 73% of the total land-to-water conversion. Conversions to agriculture have mainly been occurred from grasslands and sparsely vegetated areas as large as 1,314.95 and 1,325.84 ha, respectively. Urban coverage doubled in this period as a result of 1,443.45 ha of increase. Urban area increased in the second period from 2,920 to 3,526 ha. Conversions to agriculture occurred at high amounts. A total of 1,075.79 ha area changed from sparsely vegetated areas to crop fields. A landscape-level environmental monitoring scheme based on satellite remote sensing was proposed for effective environmental resource management. PMID- 18398691 TI - Surface ozone and meteorological condition in a single year at an urban site in central-eastern China. AB - Surface ozone and some meteorological parameters were continuously measured from June 2003 to May 2004 at urban Jinan, China. The levels and variations of surface ozone were studied and the influences of meteorological parameters on ozone were analyzed. Annual and diurnal ozone variation patterns in Jinan both show a typical pattern for polluted urban areas. Daytime ozone concentrations in summer were the highest in the four seasons. However, during nighttime from 2100 to 0600 hours ozone concentrations in spring was higher than that in summer. Daily averaged ozone showed negative correlation with pressure and relative humidity and positive correlation with temperature, total solar radiation, sunshine duration and wind speed during the study period. Further studies show that, solar radiation is a primary influence factor for the daytime variations of ozone concentrations at this site; transport of pollutants by wind could enhance the pollution at this site; precipitation has a significant influence on decreasing surface ozone. A multi-day ozone episode from 16 to 21 June 2003 was observed at this site. Surface meteorological data analysis and backward trajectory computation show that the episode is associated with the influence of typhoon Soudelor, attributing to both local photochemical processes and transport of air pollutants from southeastern coastal region, especially Yangtze River Delta region. PMID- 18398692 TI - Detection of amount and activity of living algae in fresh water by dehydrogenase activity (DHA). AB - A study was performed to determine the amount and activity of living algae in fresh water by measuring the dehydrogenase activity (DHA) of algae in order to provide a method to assess the effect of algicide treatment. The conditions of measurement were researched with respect to incubating temperature and duration, and selection of extractants. The comparison between this method and an alternative method, chlorophyll a, shows that this method is simple and easy to practice, and can determine the effect of algicide treatment. PMID- 18398693 TI - Screening of brick-kiln area soil for determination of heavy metal Pb using LIBS. AB - Rapid measurement of heavy metals in soil is an important factor in modeling the effect of industrial pollution on agricultural soil. Conventional methods of heavy metal analysis are relatively slow in terms of measurement/analysis time and sample preparation time with the requirement of skilled manpower. Our results highlight the quantitative analysis of toxic metal lead (Pb), for the first time, in an Indian agricultural soil, in the vicinity of brick-kiln area, Phaphamau, near Allahabad, India, by using a novel technique named as Laser-induced breakdown spectroscopy (LIBS). LIBS spectra of soil has been recorded in the wavelength range from ultraviolet (UV) to infrared region (200-1,100 nm). The suitability of Pb lines for drawing the calibration curve is checked and realized, for the first time, that 220.3 nm, which is observed in the UV region of LIBS spectra, is completely interference free and best suited for the quantification of trace amount of Pb in soil instead of any other Pb lines, because it has best linear regression coefficient and smallest standard deviation of the background signal. In the present work the detection limit for Pb in soil is found to be 45 ppm. Based on the present work the concentration of Pb in agricultural soil of brick-kiln area in Phaphamau is found to be congruent with 570 ppm, which is more than the regulatory standards imposed by US Environmental Protection Agency (400 ppm) for the presence of lead in soil, therefore, it is of great concern to us. PMID- 18398694 TI - Percutaneous nephrolithotomy (PCNL) in patients with previous open stone surgery. AB - OBJECTIVES: Our objective was to investigate the effects of previous open nephrolithotomy on technical features, outcomes, and morbidities of subsequent percutaneous nephrolithotomy (PCNL). METHODS: One hundred and sixty patients underwent PCNL between December 2004 and September 2006. The patients were divided into those who had previous open nephrolithotomy on the same kidney (group 1: 55 patients) and those who had no previous open surgery (group 2: 105 patients). Technical features encountered during operation and outcomes were compared between groups. RESULTS: There were no significant differences between groups with respect to mean age (group 1: 42.6 +/- 10 years vs. group 2: 45.5 +/- 9.6 years), body mass index (24.8 +/- 2.11 vs. 24.6 +/- 2.14), and stone burden (385.6 +/- 140.6 mm(2 )vs. 401.05 +/- 142 mm(2)). In group 1, 28 and 27 stones were located in the right and left kidney, respectively, whereas the location was 51 and 54 for the right and left kidney, respectively, in group 2. When the groups were compared, the mean operative time was significantly longer (155 +/- 30 min vs. 137 +/- 30 min) in group 1. But there was no significant difference with respect to requirement for secondary intervention (11% vs. 10%). Sepsis developed in two patients in group 1 and one patient in group 2. One patient in group 1 died due to septic shock. Ten (18.2%) patients in group 1 and 13 (12.4%) patients in group 2 required blood transfusion. CONCLUSIONS: When PCNL is performed after previous open nephrolithotomy, the operative time lengthens. But there is no difference with respect to success rate and morbidities. PMID- 18398695 TI - Genetic polymorphisms of GSTO2, GSTM1, and GSTT1 and risk of gastric cancer. AB - OBJECTIVE: The glutathione S-transferases (GSTs) are a superfamily of proteins that participates in detoxification. The GSTs were dividing into several classes including omega (GSTO), micro (GSTM) and theta (GSTT) classes. In human GSTO2, GSTM1, and GSTT1 are polymorphic. In order to study whether GSTO2, GSTM1, and GSTT1 polymorphisms are associated with increased gastric cancer risk in Iranian patients, the present case-control study was done. METHODS: Genomic DNA was extracted from peripheral blood of 67 gastric cancer patients and 134 control subjects. The genotyping was performed using PCR-based method. The possible association of gastric cancer with the GSTO2 N142D polymorphism was estimated with assuming additive, dominant, and recessive effect of the variant 142D allele. To investigate whether profiles of GST genotypes are associated with the risk of gastric cancer, we used unconditional logistic regression analysis. RESULTS: The GSTO2 142D allele in additive, dominant and recessive models was not associated with the risk. Because GSTM1, GSTT1, and GSTO2 genes belong to low penetrance genes which might be involved in the carcinogenesis, patients and controls without family of cancer in first-degree relatives were also analyzes separately. To investigate whether profiles of GST genotypes are associated with the risk of gastric cancer, we used unconditional logistic regression analysis with GSTM1, GSTT1, and GSTO2 genotypes as predictor factors. The GSTO2 DD genotype was associated with decreased risk as compared to GSTO2 NN genotype (OR = 0.21, 95% CI: 0.05-0.92, P = 0.038). CONCLUSIONS: Present findings show that GSTO2 DD genotype decreases the risk of gastric cancer in individuals without history of cancer in their first-degree relatives. PMID- 18398696 TI - Communicating genetic information in the family: enriching the debate through the notion of integrity. AB - Genetic information about one individual often has medical and reproductive implications for that individual's relatives. There is a debate about whether policy on transmitting genetic information within the family should change to reflect this shared aspect of genetic information. Even if laws on medical confidentiality remain unchanged, there still remains the question of professional practice and whether, to what extent and by what means professionals should encourage disclosure within a family. The debate so far has tended to focus on who has a right to genetic information, or has a right to decline genetic information, frequently drawing on the notion of individual autonomy. There are significant divergences within this debate, and difficulties with the use of autonomy in this context have been noted. This paper draws on theoretical considerations as well as on qualitative empirical data to show that shifting from talk of autonomy to talk of integrity will greatly enrich and illuminate the issues. It becomes possible to gain deeper insights into the ethical significance of the timing and the manner of such communication, the character of recipients of knowledge, and the nuanced nature of communication and different levels of understanding within a family. PMID- 18398697 TI - Types of centredness in health care: themes and concepts. AB - BACKGROUND: For a variety of sociological reasons, different types of centredness have become important in health and social care. In trying to characterize one type of centredness, we were led to consider, at a conceptual level, the importance of the notion of centredness in general and the reasons for there being different types of centeredness. METHOD: We searched the literature for papers on client-, family-, patient-, person- and relationship- centred care. We identified reviews or papers that defined or discussed the notions at a conceptual level. The reviews and papers were analyzed as text transcripts. RESULTS: We identified 10 themes that were common to all the types of centredness. At a conceptual level we could not identify thematic differences between the types of centredness. These findings were subjected to a philosophical critique using ideas derived from Wittgenstein. CONCLUSION: Different types of centredness are required in different contexts. The differences are justified by their practical utility. The unifying themes of centredness, however, reflect a movement in favour of increasing the social, psychological, cultural and ethical sensitivity of our human encounters. PMID- 18398698 TI - Abortion, embryonic stem cell research, and waste. AB - Can one consistently deny the permissibility of abortion while endorsing the killing of human embryos for the sake of stem cell research? The question is not trivial; for even if one accepts that abortion is prima facie wrong in all cases, there are significant differences with many of the embryos used for stem cell research from those involved in abortion--most prominently, many have been abandoned in vitro, and appear to have no reasonably likely meaningful future. On these grounds one might think to maintain a strong position against abortion but endorse killing human embryos for the sake of stem cell research and its promising benefits. I will argue, however, that these differences are not decisive. Thus, one who accepts a strong view against abortion is committed to the moral impermissibility of killing human embryos for the sake of stem cell research. I do not argue for the moral standing of either abortion or the killing of embryos for stem cell research; I only argue for the relation between the two. Thus the conclusion is relevant to those with a strong view in favor of the permissibility of killing embryos for the sake of research as much as for those who may strongly oppose abortion; neither can consider their position in isolation from the other. PMID- 18398699 TI - Individual quality of life: adaptive conjoint analysis as an alternative for direct weighting? AB - In the schedule for the evaluation of individual quality of life (SEIQoL) the weights for five individualized quality of life domains have been derived by judgment analysis and direct weighting (DW). We studied the feasibility and validity of adaptive conjoint analysis (ACA) as an alternative method to derive weights in 27 cancer patients and 20 patients with rheumatoid arthritis. Further, we assessed the convergence between direct weights and weights derived by ACA, and their correlation with global quality-of-life scores. All respondents finished the ACA task, but one in five respondents were upset about the ACA task. Further, the task was vulnerable to judgment 'errors', such as inconsistent answers. The agreement between the two weights was low. Both weighted index scores were strongly correlated to the unweighted index score. The relationships between the index score and scores on a visual analogue scale for global individual quality of life and global quality of life were similar whether or not the index score was calculated with DW weights, with ACA weights, or without using weights. We conclude that, because weights did not improve the correlation between the index score and global quality of life scores, it seems sufficient to use the unweighted index score as a measure for global individual quality of life. PMID- 18398700 TI - Hepatitis C virus ARFP/F protein interacts with cellular MM-1 protein and enhances the gene trans-activation activity of c-Myc. AB - The ARFP/F protein is synthesized from the +1 reading frame of the hepatitis C virus (HCV) core protein gene. The function of this protein remains unknown. To study the function of the HCV ARFP/F protein, we have conducted the yeast two hybrid screening experiment to identify cellular proteins that may interact with the ARFP/F protein. MM-1, a c-Myc interacting protein, was found to interact with HCV ARFP/F protein in this experiment. The physical interaction between ARFP/F and MM-1 proteins was further confirmed by the GST pull-down assay, the co immunoprecipitation assay and confocal microscopy. As MM-1 can inhibit the gene transactivation activity of c-Myc, we have conducted further analysis to examine the possible effect of the ARFP/F protein on c-Myc. Our results indicate that the HCV ARFP/F protein can enhance the gene trans-activation activity of c-Myc, apparently by antagonizing the inhibitory effect of MM-1. The ability of the ARFP/F protein to enhance the activity of c-Myc raises the possibility that ARFP/F protein might play a role in hepatocellular transformation in HCV patients. PMID- 18398701 TI - Expression and membrane integration of SARS-CoV M protein. AB - SARS-CoV M gene fragment was cloned and expressed as a recombinant protein fused with a V5 tag at the C-terminus in Vero E6 cells. In addition to un-glycosylated and glycosylated proteins, one product with smaller size initiated in-frame from the third Met residues probably through ribosomal re-initiation was also detected. Translation initiated in-frame from the third Met is unusual since the sequence around the first Met of SARS-CoV M protein contains the optimal consensus Kozak sequence. The function of this smaller translated product awaits further investigation. Similar to other N-glycosylated proteins, glycosylation of SARS-CoV M protein was occurred co-translationally in the presence of microsomes. The SARS-CoV M protein is predicted as a triple-spanning membrane protein lack of a conventional signal peptide. The second and third trans-membrane regions (a.a. 46-68 and 78-100) are predicted to be the primary type helices, which will be able to penetrate into membrane by themselves, while the first trans-membrane region (a.a. 14-36) is predicted to be the secondary type helix, which is considered to be stabilized by the interaction with other trans-membrane segments. As expected, the second and third trans-membrane regions were able to insert a cytoplasmic protein into the endoplasmic reticulum membrane more efficiently than the first one. These results should be important for the study of SARS-CoV morphogenesis. PMID- 18398702 TI - The determination of VEGF and MVD, among patients with primary breast cancer. AB - The purpose of the study was to ascertain the value of assessment of vascular endothelial growth factor (VEGF) levels and microvessel density, and to search for correlations between them, in women with breast cancer. The assessment considered factors such as the stage of clinical disease advancement--according to International Union Against Cancer, the grade of histological malignancy, status of axillary lymph nodes and the size of the primary tumour. The concentration of VEGF was assessed in the plasma of 103 women with breast cancer, using an immunoenzymatic method (Quantikine test of R&D Systems). Assessment of microvessel density was performed using histopathological immunoperoxidase methods, using an anti-von Willebrand factor antibody (DAKO A/S). A statistically significant relationship was found between rising VEGF levels and microvessel density in women with breast cancer, when compared to values from a control group. A correlation was found between VEGF concentration and microvessel density (MVD) values. Statistically significant differences were found between VEGF levels of patients in stages I, II and III of clinical disease advancement. For MVD, differences were found only between stages I and III. A statistical relationship was also found between VEGF and MVD and tumour size. Similar results were found between VEGF concentrations in women with metastases to the axillary lymph nodes and cytokine levels in women with no metastases. The results of the study suggest that the degree of tumour vascularization and the concentration of VEGF may represent valuable indicators for the assessment of the angiogenic process in women with breast cancer. PMID- 18398704 TI - In vitro effects of preservative-free tafluprost and preserved latanoprost, travoprost, and bimatoprost in a conjunctival epithelial cell line. AB - PURPOSE: This study compared the toxicity profiles of three antiglaucoma prostaglandin F2alpha analogs, latanoprost, travoprost, and bimatoprost which contain benzalkonium chloride (BAK), with tafluprost, a new preservative-free prostaglandin analog. METHODS: IOBA-NHC cells were exposed to BAK-containing prostanoid solutions, their respective BAK concentrations, and preservative-free tafluprost solution for 30 min. Membrane integrity, apoptosis, oxidative stress, and cells morphology were evaluated. RESULTS: Preservative-free tafluprost resulted in significantly higher membrane integrity and lower pro-apoptotic and pro-oxidative effects than preservative-containing prostaglandin analog preparations. CONCLUSIONS: These results suggest that tafluprost, a new preservative-free prostaglandin analog, has very low or no pro-apoptotic, pro necrotic, or pro-oxidative effects in vitro compared to preservative-containing formulations. PMID- 18398703 TI - P53 and BCL-2 as prognostic markers in endometrial carcinoma. AB - The objective of this study was to verify the frequency of p53 and BCL-2 immunohistochemical expression in patients with endometrial carcinoma and to correlate it with histological factors (histological type, tumor grade, depth of myometrial invasion, lymph node involvement and surgical staging) and survival. Forty-eight patients with endometrial carcinoma who were submitted to primary surgical treatment were assessed. p53 and BCL-2 immunohistochemical expression was determined using paraffin blocks containing the tumor area. p53 and BCL-2 expression was detected in 39.6% and 58.3% of the tumors, respectively. No significant difference was found regarding the frequency of p53 expression when analyzing histological type (33.3% in endometrioid tumors, 58.3% in non endometrioid tumors; p = 0.176), depth of myometrial invasion (p = 0.632) and surgical staging (I-11.1%, II-66.7%, III-57.1%; p = 0.061). p53 expression was significantly more frequent in undifferentiated tumors (p = 0.007) and in those showing lymph node involvement (p = 0.030). Univariate analysis showed a positive association with death (RR, 3.358; CI, 1.386-8.134; p = 0.005) and short-term survival. The present study did not reveal any correlation between BCL-2 expression and histopathologic markers or survival. In conclusion, this study showed that p53 expression is directly correlated with undifferentiated tumors, lymph-node involvement and risk of death. On the other hand, BCL-2 expression was not correlated with any known histological factors. PMID- 18398705 TI - Novel CYP4V2 gene mutation in a Mexican patient with Bietti's crystalline corneoretinal dystrophy. AB - PURPOSE: To report the clinical and genetic analysis of a Mexican female patient with a sporadic Bietti's crystalline corneoretinal dystrophy. METHODS: Ophthalmological examination included best-corrected visual acuity, slit lamp examination, applanation tonometry, fundus photography, fluorescein retinal angiography, Goldmann kinetic perimetry, corneal rotating Scheimpflug imaging, and anterior segment optical coherence tomography (Visante OCT). Genetic analysis included PCR amplification and direct nucleotide sequencing of the entire CYP4V2 gene in DNA from the propositus and her relatives. RESULTS: A late-stage retinal dystrophy was established in the patient. No retinal or corneal crystalline deposits were evident during clinical evaluation. Retrospective analysis of fundus imaging disclosed the presence of retinal crystalline deposits, suggesting the diagnosis of Bietti's crystalline corneoretinal dystrophy. Molecular analysis of the CYP4V2 gene revealed the presence of a novel C to T mutation at nucleotide position 974 (exon 7), predicting a threonine to isoleucine replacement at amino acid position 325. Corneal deposits were not seen by biomicroscopy, corneal OCT, or specular microscopy but were evidenced by means of the corneal rotating Scheimpflug imaging. CONCLUSION: Our results expand the allelic heterogeneity of Bietti's crystalline corneoretinal dystrophy. This is the first patient of Latin American origin in which a molecular analysis of the disease has been performed. Our results suggest that the use of corneal rotating Scheimpflug imaging can evidence corneal deposits that are not apparent by other methods. PMID- 18398706 TI - Expression of toll-like receptors in the Fusarium solani infected cornea. AB - PURPOSE: To investigate the expression of toll-like receptors (TLRs) in the human cornea with Fusarium solani (F. solani) keratitis. METHODS: Five human corneas with F. solani keratitis and 5 healthy human corneas were evaluated for TLR1-10 mRNA by reverse transcribed polymerase chain reaction (RT-PCR) and relative real time PCR. The proteins of TLR2, 4, and 9 were also compared by Western blot. The mean times between these corneas were collected, and the onset of treatment to surgery was 34.4 +/- 12.4 days, ranging from 15 to 47 days. The data were analyzed with SPSS15.0. RESULTS: TLR1-10 mRNA was expressed in both healthy and F. solani infected human corneas. The expression of TLR2, 4, 6, and 9 mRNA in the F. solani infected human corneas were upregulated. The Western blot showed the protein expression of TLR2, 4, and 9 was also upregulated in the corneas with F. solani keratitis than that of the healthy corneas. CONCLUSIONS: TLRs are expressed diversely in the F. solani infected human cornea, and TLR2, 4, and 9 may be implicated in the pathogenesis of F. solani infection in the cornea. PMID- 18398707 TI - Is there a relationship between blood pressure and intraocular pressure? An experimental study in hypertensive rats. AB - PURPOSE: Evaluation of relation between blood pressure (BP) and intraocular pressure (IOP) in two hypertensive rat strains: spontaneously hypertensive rats (SHR) and double transgenic (dTGR) (harboring human renin and angiotensinogen genes) rats, and in their normotensive control Wistar Kyoto and Sprague Dawley rats, respectively. METHODS: Each rat strain was divided into medicated and non medicated groups. Medicated rats were treated orally with an angiotensin II receptor type 1 blocker. IOP was measured using a specific rebound tonometer and BP by a tail-cuff method. Both parameters were determined in conscious animals every second week. For comparison, at the end of the study, IOP was measured in conscious and anesthetized rats. RESULTS: The baseline IOP was higher in hypertensive rats vs their normotensive controls. Eight weeks of treatment with an angiotensin type 1 receptor blocker did not prevent a slight increase in IOP, although it abolished the development of hypertension in SHR. The markedly elevated IOP was reduced in medicated and non-medicated dTGR animals during the short follow-up period. General anesthesia reduced IOP significantly. CONCLUSION: The results suggest a positive relation between BP and IOP in hypertensive rats. PMID- 18398708 TI - Evaluation of the modulation of P-glycoprotein (P-gp) on the intraocular disposition of its substrate in rabbits. AB - PURPOSE: To evaluate the functional role of P-gp and ocular tissue distribution of intravitreally injected Rhodamine-123 (Rho-123) in the presence of P-gp specific blocker (GF 120918) in normal as well as rifampicin-fed rabbits using microdialysis and direct sampling technique. METHODS: Intravitreal pharmacokinetics of Rho-123 were conducted in male New Zealand albino rabbits. Direct sampling and microdialysis were employed to study the disposition of Rho 123 in normal as well as rifampicin-fed conditions. Control animals received Rho 123 at the concentration of 350 ng in PBS (0.05 ml) intravitreally, and the blocker-treated group received GF 120918 intravenously at the dose of 3.5 mg/kg 30 min prior to intravitreal injection of Rho-123. In case of direct sampling, four eyes were enucleated at different time points, and ocular tissues and humors were stored at -86 degrees C until analysis by HPLC with fluorescence detection. RESULTS: In direct sampling, the blocker group showed significant increase (2.6 fold) in the mean vitreous concentration of Rho-123. Other tissues like ret choroid, iris, and cornea also showed significant increase in their mean concentration. Microdialysis did not significantly predict the changes observed with direct sampling. Rifampicin-fed rabbits showed a vitreous pharmacokinetic profile comparable with non-fed (control) animals, and the pharmacokinetic parameters were unaffected by the blocker pretreatment. CONCLUSION: Intravenously injected blocker significantly altered the ocular disposition of intravitreally injected P-gp substrate. Rifampicin pretreatment did not upregulate P-gp transporters of the retina to the extent to affect the intravitreal kinetics of Rho-123 significantly. PMID- 18398709 TI - An assessment of intraocular pressure change in healthy subjects during air flight. AB - PURPOSE: To investigate the effect of air flight on intraocular pressure (IOP). METHODS: Baseline IOP of 25 healthy volunteers was measured at a ground level of 1760 feet above sea level (ASL) using Tono-Pen XL. Measurements were repeated after reaching an altitude of 19,000 feet and then on the second hour during a routine flight. Cabin pressure was kept around 8000 feet. IOP measurement was repeated after landing (3.5 hr after taking off). Change in the IOP was evaluated. RESULTS: Mean +/- SD IOP was 14.2 +/- 2.7 mmHg at ground level and was 14.0 +/- 2.2 mmHg after gaining maximum altitude (p = 0.78). Mean IOP dropped to 12.3 +/- 2.5 mmHg on the second hour of flight and 12.0 +/- 1.7 mmHg after landing. Decrease in IOP on the second hour (13.4%, p = 0.005) and after landing (15.7%, p = 0.001) were significant. CONCLUSION: IOP significantly decreased on the second hour measurements during a routine flight, and this change was sustained after landing. PMID- 18398710 TI - Femtosecond laser-assisted retinal imaging and ablation: experimental pilot study. AB - PURPOSE: To investigate retinal imaging and ablation using femtosecond laser pulses. MATERIALS AND METHODS: Two non-amplified near-infrared femtosecond lasers were used to irradiate porcine retinal specimens in vitro. The lasers were used for tissue removal as well as multiphoton laser scanning microscopy. RESULTS: Ablation of the nerve fiber layer was performed at pulse energies of 1.0 nJ to 3.9 nJ. Control laser scanning images were acquired within seconds after irradiation. Specimens were additionally investigated with electron microscopy. CONCLUSIONS: Non-amplified femtosecond lasers may allow precise surgery controlled by fast high-resolution imaging of the target. PMID- 18398711 TI - Natural history and histology in a rat model of laser-induced photothrombotic retinal vein occlusion. AB - PURPOSE: To observe temporal changes in retinal physiology and histology in a rat model of laser-induced retinal vein occlusion (RVO). METHODS: Ophthalmoscopy, fundus photography, and fluorescein angiography (FA) were performed following laser-induced central retinal vein occlusion (CRVO) and branch retinal vein occlusion (BRVO) immediately after laser treatment and at 3 and 6 hr and 2, 4, 7, 14, and 21 days. Retinal histology was examined at 4, 7, 14, and 21 days. Full field electroretinogram was recorded from both eyes simultaneously at day 4. RESULTS: For CRVO and BRVO, reperfusion of occluded branch veins was observed 1 to 2 days after treatment. Despite complete reperfusion of branch veins, retinal edema and hemorrhages peaked on day 4, and by day 14, treated retinas appeared pale and edematous upon ophthalmoscopy. In BRVO animals, retinal hemorrhages were limited to the vein-occluded region, although edema was more widespread and, to a limited extent, involved the untreated hemi-retina. Significant GCL cell loss was observed in both CRVO and BRVO groups after day 14. Regional analysis showed that relative GCL loss was greatest in the peripheral retina in BRVO group. Electroretinography disclosed moderate to severe functional deficits in photoreceptors, bipolar, and amacrine and ganglion cells. CONCLUSION: Laser induced RVO in rats results in targeted vascular occlusion that persisted for 1 to 2 days. Functional deficits were evident and significant GCL cell loss was seen, notably within peripheral retina of the BRVO model. This reproducible model provides a valuable tool for the study of the molecular events associated with retinal ischemia and cell death. PMID- 18398712 TI - No association found between the promoter variants of TNF-alpha and diabetic retinopathy in Chinese patients with type 2 diabetes. AB - PURPOSE: Necrosis tumor factor (TNF) is known to be associated with diabetic retinopathy (DR). The objective of this study was to examine the genetic variations of TNF-alpha and assess their possible relationship to DR in type 2 diabetic patients in the Chinese population. MATERIALS AND METHODS: We conducted a case-control association study between the promoter variants of TNF-alpha and diabetic retinopathy in Chinese patients with type 2 diabetes. We selected three variants in the promoter region of TNF-alpha, namely rs1800629, rs1041981, and rs2857713. RESULTS: No individual SNP nor any haplotype was found to be associated with DR in our study. CONCLUSION: This is the first study to report TNF-alpha polymorphisms in patients with DR in the Chinese population. The results suggest that the variants among the promoter of TNF-alpha are unlikely to play a major role in the susceptibility to DR in Chinese patients with type 2 diabetes. PMID- 18398713 TI - The role of alkylphosphocholines in retinal Muller glial cell proliferation. AB - PURPOSE: Alkylphosphocholines (APCs) are investigated for their effect on Muller glial cell proliferation and F-actin stress fiber distribution in vitro. MATERIALS AND METHODS: Muller cells were incubated with APCs (C18:1-PC and C22:1 PC) +/- fetal calf serum. Proliferation was assessed by BrdU labeling and with the tetrazolium dye-reduction assay. Toxicity was measured using the trypan blue exclusion assay. The distribution of F-actin stress fibers was determined using FITC-phalloidin staining. RESULTS: APCs are effective inhibitors of human and rat Muller glial cell proliferation and hypoxia-induced up-regulation of F-actin stress fibers in vitro in non-toxic concentrations. CONCLUSIONS: APCs might prevent intraretinal changes as a result of serum stimulation and hypoxia following retinal detachment. PMID- 18398714 TI - Increased plasma and optic nerve levels of IL-6, TNF-alpha, and MIP-2 following induction of ischemic optic neuropathy in mice. AB - PURPOSE: To investigate levels of proinflammatory cytokines in a mouse model of anterior ischemic optic neuropathy (rAION). METHODS: AION was induced in C57/BL6 mice and levels of IL-6, TNF-alpha, and MIP-2 were measured in plasma by ELISA and in the optic nerves by RT-PCR at predetermined intervals. RESULTS: Plasma: IL 6 levels were elevated immediately after rAION induction and decreased gradually thereafter. TNF-alpha showed an early peak on day 1 and again from day 21. MIP-2 levels were increased until day 7. Optic nerve: IL-6, TNF-alpha, and MIP-2 levels increased within a few hours and then decreased gradually. IL-6 had a second peak on day 3. CONCLUSIONS: Proinflammatory cytokines may play a role in the pathogenesis of rAION. PMID- 18398717 TI - Requirement of lipocalin 2 for chronic myeloid leukemia. AB - The Bcr-Abl oncoprotein causes chronic myeloid leukemia by a mechanism involving its activated tyrosine kinase. BCR-ABL+ mouse hematopoietic cells persistently express and secrete lipocalin 2 by a mechanism that requires the tyrosine kinase of the Bcr-Abl oncoprotein. Our new findings indicate that lipocalin 2 is required for leukemia induction, as prevention of expression of lipocalin 2 by BCR-ABL+ mouse marrow cells totally blocks leukemia induction in a mouse model. PMID- 18398718 TI - Clinical perspectives of concepts on neoplastic stem cells and stem cell resistance in chronic myeloid leukemia. AB - In chronic myeloid leukemia (CML), neoplastic stem cells and/or their subclones exhibit resistance against BCR/ABL tyrosine kinase inhibitors (TKIs). Therefore, residual CML stem cells (subclones) in TKI-treated patients are a logical target of therapy, and their elimination is considered a major aim and clue in the development of curative treatment approaches. A number of different mechanisms may underly resistance of CML stem cells against TKIs and other targeted or/and conventional drugs, including stem cell quiescence, expression of drug transporters, stem cell plasticity, BCR/ABL mutations, overexpression of BCR/ABL and BCR/ABL-independent signalling- and survival-molecules. In this article, possibilities to overcome stem cell resistance in CML by exploiting knowledge on molecular mechanisms that underly the 'stem cell escape' from drug therapy are discussed. PMID- 18398719 TI - BCR/ABL, DNA damage and DNA repair: implications for new treatment concepts. AB - BCR/ABL fusion tyrosine kinase transforms hematopoietic stem cells causing chronic myelogenous leukemia (CML) and acute lymphoblastic leukemia (ALL). BCR/ABL regulates numerous proteins involved in apoptosis, proliferation and cell - cell or cell - extracellular matrix interactions. However, BCR/ABL also enhances DNA damage caused by endogenous reactive oxygen species and exogenous genotoxic treatment. In addition, BCR/ABL modulates the response to DNA damage to promote genomic instability. This function leads to resistance to ABL kinase small molecular inhibitors (SMIs) imatinib (IM), dasatinib and nilotinib, and contributes to malignant progression of the disease. The former phenomenon is often caused by mutations in BCR/ABL kinase whereas the latter is associated with accumulation of additional genetic aberrations including chromosomal translocations, deletions, additional chromosomes, gene amplifications, and point mutations. Possible benefits of anti-mutagenic therapy used in pursuing the cure of BCR/ABL-positive leukemias are discussed. PMID- 18398720 TI - Target spectrum of the BCR-ABL inhibitors imatinib, nilotinib and dasatinib. AB - Following the initial success of imatinib as frontline therapy for chronic myeloid leukemia (CML), several second-generation therapeutics have been developed with increased potency and the ability to inhibit the majority of imatinib-resistant mutations. Here, we review the current knowledge about the target specificity of the two new inhibitors nilotinib and dasatinib in comparison to imatinib, including the recent large-scale chemical proteomics screens. PMID- 18398721 TI - Signal interception-based therapies--a double-edged sword in Bcr/abl-induced malignancies? AB - Imatinib was a major breakthrough in the treatment of Bcr/abl-positive leukemias. The effectiveness and value of this drug is limited by the emergence of resistance. Alternative drug targets may be identified by analyzing the downstream signaling network including the Jak/Stat-pathway, Ras-dependent signaling, PI3-kinases (PI3K), or the nuclear transcription factors onto which these pathways impinge. However, several factors limit the possible suitability of a drug target: (i) tissue-specific versus ubiquitous expression of the target; (ii) redundancy within the signaling network; and (iii) off-target effects on the immune system. Although the former two aspects are well appreciated as limiting factors, the latter has not been addressed so far. The advent of genetically engineered mice provides a sophisticated target validation in vivo as well as analysis of interactions between the immune system and tumor cells. Based on studies in such mouse models, we predict that many targeted compounds including PI3Kdelta-inhibitors, could act as double-edged swords because their beneficial action on tumor cells may be neutralized or even overwhelmed by their additional immunosuppressive effects. PMID- 18398722 TI - New therapeutic approaches and prognostic factors in chronic myeloid leukemia. AB - Imatinib mesylate is now the first-choice treatment for all newly diagnosed CML patients, but despite the impressive percentage of responding patients, some CML cases show primary resistance or relapse after an initial response. Although some clinical and biological findings have been found to be associated with a lower probability of response to imatinib, at present no precise markers allowing to predict outcome for individual patients exist. The most common mechanisms of resistance to imatinib include BCR-ABL kinase domain mutations, BCR-ABL amplification and overexpression, and clonal evolution with activation of additional transformation pathways. These mechanisms are caused by the genomic instability, which characterises the Ph-positive clone. Several approaches to overcome resistance have been proposed, including novel tyrosine kinase inhibitors (TKIs) that have now reached the clinical or pre-clinical phase of evaluation. Clinical trials with these novel TKIs have shown good rates of hematologic and cytogenetic responses that appear also durable in time, but still cases of resistance are also observed, supporting the notion that genomic instability represents the major determinant affecting the final outcome of the CML patients when treated with TKIs. Understanding exactly the mechanisms leading to genomic instability of the Ph-positive cells represents therefore the real challenge for the near future. PMID- 18398723 TI - Immunotherapeutic approaches in chronic myelogenous leukemia. AB - Chronic myeloid leukemia (CML) is a stem cell disease in which BCR/ABL plays an important role as oncoprotein and also as a molecular and immunogenic target. In fact, several molecular targeted drugs and immunotherapies have been developed for this disease. In the current article, several immunotherapeutic approaches in CML, including interferon-alpha and vaccination, are discussed along with results from clinical trials and the value of such immunotherapies in the imatinib-era. PMID- 18398724 TI - Diagnostic and prognostic value of new biochemical and immunohistochemical parameters in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is a stem-cell disease characterized by multilineage expansion of clonal BCR/ABL+ cells. Transformation from chronic into accelerated and blast phase of CML is usually associated with drug resistance and is accompanied by typical clinical and/or laboratory features, such as splenomegaly, increase in precursor cells, disturbed megakaryopoiesis, basophilia or marrow fibrosis. Because of new treatment options, early recognition of disease-acceleration is of importance. In this article, we review most recent developments in diagnostic procedures employing basophil-related biochemical and histopathological markers. These tests are useful to quantitate basophil-lineage cells in the peripheral blood in CML, to determine and quantify basophilia in the bone marrow, and to detect focal accumulations of blast cells and megakaryocytes as well as increased angiogenesis and fibrosis in bone marrow sections. Application of these markers may assist in determining the phase of disease and may help to better predict the prognosis in individual patients. PMID- 18398725 TI - Pharmacologic markers and predictors of responses to imatinib therapy in patients with chronic myeloid leukemia. AB - Clinical resistance to imatinib often occurs in the absence of a mutation in the BCR-ABL kinase domain. Imatinib is transported out of cells by the efflux transporter ABCB1 (MDR1, whose product is p-glycoprotein). By contrast, the influx transporter, human organic cation transporter 1 (hOCT1) (also known as SLC22A1), transports imatinib into cells. Recent studies have identified that patients with low expression or activity of hOCT1 have a lower probability of achieving a cytogenetic or molecular remission. Prospective studies are currently investigating whether early trends in transporter expression can be used to guide treatment decisions. Plasma imatinib levels are higher in patients responding well to treatment, and may be useful in patients with suboptimal response or dubious compliance. Uptake of the second generation tyrosine kinase inhibitors, dasatinib and nilotinib, is less dependent upon hOCT1. These two drugs may therefore achieve adequate intracellular concentrations even in patients with low hOCT1 expression. PMID- 18398726 TI - Possible role for gene expression profiling in predicting responses to conventional or targeted drugs in patients with chronic myeloid leukemia. AB - The introduction of imatinib has profoundly changed the clinical management and prognosis of patients with chronic myeloid leukemia (CML). However, prognostic scores introduced during the interferon-era have lost their prognostic value when applied to patients treated with imatinib. Gene expression profiling represents a unique and powerful tool for the identification of responders and non-responders to cancer therapy. Thus, this technology may also represent a valuable strategy to identify patients with insufficient response prior to or during imatinib treatment, who might benefit from alternative treatment modalities. This review will briefly summarize the recently published data on gene expression profiling for imatinib response prediction in CML patients. PMID- 18398727 TI - Chronic lymphocytic leukemia with mutated IgVH1-69 usage: a distinct CLL subgroup with more indolent behavior? PMID- 18398728 TI - DNA methyltransferase inhibitors: class effect or unique agents? PMID- 18398729 TI - The development of novel immunotherapeutic approaches in multiple myeloma. PMID- 18398730 TI - Mantle, blastic, Burkitt. PMID- 18398731 TI - Bortezomib: putting mantle cell lymphoma on death row. PMID- 18398732 TI - Early interim PET scan in Hodgkin lymphoma: where do we stand? AB - Positron emission tomography using [18F]-fluoro-2-deoxy-d-glucose (FDG-PET) is an important tool for staging and treatment response assessment in malignant lymphomas. In Hodgkin lymphoma (HL), FDG-PET precisely predicts the therapy response when performed very early during standard ABVD chemotherapy. However, it is unclear whether FDG-PET retains this role if therapy is changed as a consequence of the scan, or if performed during a more intensive chemotherapy regimen such as BEACOPPesc, which is used for HL. This brief review presents the up-to-date evidence for the use and interpretation of early interim FDG-PET in HL, including recent preliminary results on early interim FDG-PET during BEACOPPesc therapy. PMID- 18398733 TI - Current treatment approaches for diffuse large B-cell lymphoma. AB - There have been two major developments over the last decade that has led to improvements in outcome and longer survival for patients with diffuse large B cell lymphoma (DLBCL). These developments have been firstly to increase the dose of active cytotoxic drugs and shorten the time between cycles, resulting in dose dense and/or dose-intense regimens and secondly the addition of the anti-CD20 monoclonal antibody rituximab to chemotherapy. Both strategies have been associated with higher response rates, lower relapse rates, longer event-free survival (EFS) and improved overall survival (OS), particularly in better prognostic groups. A combination of dose-dense and dose-intense chemotherapy regimens plus rituximab is currently being tested to confirm that the use of both approaches confers survival advantage. High-risk, poorer-prognosis DLBCL remains a challenge, and new treatment strategies are required for these patients. Improvements in outcome may potentially be achieved through a greater understanding of the genetic abnormalities specifically associated with poorer prognosis disease, and factors that lead to unresponsiveness to chemotherapy. The role of radiotherapy is currently less clearly defined than at anytime in the management of DLBCL and the current evidence for using radiotherapy in this disease is therefore rigorously reviewed. PMID- 18398734 TI - Novel anti-myeloma agents and angiogenesis. AB - During the last decade several novel agents have been used in the management of patients with multiple myeloma. Immunomodulatory drugs and proteasome inhibitors exert their efficacy both directly by inducing apoptosis of myeloma cells and indirectly through the interruption of the interactions between myeloma and stromal cells in the bone marrow (BM) microenvironment. These interactions are crucial for myeloma cell growth and survival. The adherence of myeloma cells to BM stromal cells leads to the overproduction of several cytokines with angiogenic properties that enhance the survival and growth of myeloma cells through paracrine and autocrine loops. The correlation of these molecules with clinical features and survival of myeloma patients supports the importance of angiogenesis in the pathogenesis of the disease and reveals these cytokines as suitable targets for the development of novel anti-myeloma therapies. This review summarises all available preclinical and clinical data for the effect of novel agents that are used in myeloma therapy, such as thalidomide, lenalidomide, bortezomib and VEGF inhibitors, on angiogenesis, which is at least partially responsible for their remarkable anti-myeloma efficacy. PMID- 18398735 TI - Activity of decitabine in patients with myelodysplastic syndrome previously treated with azacitidine. AB - Azacitidine and decitabine are the two hypomethylating agents approved by the Food and Drug Administration for the treatment of patients with myelodysplastic syndrome (MDS). The efficacy of one agent post-failure of the other is unknown. Fourteen patients with MDS post-azacitidine failure/lack of response/intolerance were treated with decitabine. Overall three patients achieved a complete remission, and one patient had hematologic improvement, for an overall response rate of 28%. Of the responders, one stopped prior 5-azacitidine owing to disease progression, two for no response and one for severe skin toxicity. Grade 3-4 drug related side-effects were minimal. Global methylation studies in patient samples showed decrease of methylation after treatment with decitabine. As in our previous studies, there was no difference in hypomethylation between responders and nonresponders. We conclude that clinically significant responses with decitabine can be seen in patients post-azacitidine failure without significant toxicity. PMID- 18398736 TI - JAK2 V617F patients with essential thrombocythemia present with clinical features of polycythemia vera. AB - Recent studies have shown that Janus tyrosine kinase 2 (JAK2) V617F mutation is found in nearly all patients with polycythemia vera (PV) and underlie the basis of PV molecular pathogenesis. Moreover, JAK2 V617F patients with essential thrombocythemia (ET) have been found to have some clinical features similar to PV. To determine whether the same is true in a different Chinese patient population, we employed Allele-specific polymerase chain reaction in combination with sequence analysis to investigate the point mutation in a series of Chinese patients with hematological malignancies. A total of 99 Chinese myeloproliferative disorder patients and 120 additional patients with acute myeloid leukemia, acute lymphoblastic leukemia and myelodysplastic syndromes were studied. The V617F mutation was detected in genomic DNA of peripheral blood samples of 16 of 23 PV patients (69.6%), 21 of 45 ET patients (46.7%) and 3 of 8 patients with idiopathic myelofibrosis (37.5%). There were striking differences in clinical features such as hemoglobin, hematocrit and neutrophils percentages between V617F positive and negative patients with ET. Hence, our data support the idea that JAK2 V617F mutation divides ET patients into two subtypes, with the V617F positive group showing phenotypic similar to that of PV. PMID- 18398737 TI - Prognostic implications of the immunophenotype in biphenotypic acute leukemia. AB - The present study retrospectively analyzed clinicopathological and clinical data from 43 adult patients with biphenotypic acute leukemia (BAL) from 11 Korean institutes. The incidence of BAL was 2.1% among acute leukemias. In terms of immunophenotype, 31 patients had myeloid plus B-lymphoid (M + B), 10 had myeloid plus T-lymphoid (M + T), one had myeloid plus B-lymphoid plus T-lymphoid (M + B + T), and one had B-lymphoid plus T-lymphoid (B + T). Patients with M + T phenotype had significantly lower CR rate (55.6% vs. 88.0%, P = 0.039) and lower overall survival (0% vs. 33.9% at 5 years, P = 0.028) than those with M + B phenotype. Our results suggest that immunophenotype has prognostic implications in adult patients with BAL. PMID- 18398738 TI - Expression of CD44, but not CD44v6, predicts relapse in children with B cell progenitor acute lymphoblastic leukemia lacking adverse or favorable genetics. AB - Although significant progress has been made in the treatment of childhood acute lymphoblastic leukemia (ALL) the prognosis following relapse is still poor. Additional prognostic indicators are needed to better target treatment and thereby improve the treatment of these patients. We have previously demonstrated an association between poor outcome and CD44v6 expression in a heterogeneous cohort of patients. Others have shown by microarray analysis that CD44 expression in diagnostic samples is linked with relapse risk. In this study, we examined CD44 and CD44v6 protein expression by flow cytometry and CD44v6 mRNA expression by quantitative RT-PCR in diagnostic samples from 97 pediatric patients with ALL. We found that CD44 protein expression was associated with disease relapse and was independent of age and WCC. In contrast, high CD44v6 expression was not associated with relapse. These findings may assist in the process of refining prognostic groups for children with ALL. PMID- 18398739 TI - FDG-PET in prediction of splenectomy findings in patients with known or suspected lymphoma. AB - Diagnostic splenectomy is frequently performed in patients with suspected or known lymphoma. We evaluated whether preoperative 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) results may correlate with splenic pathology. Of 165 patients undergoing splenectomy at the Weill Cornell Medical Centre/New York Presbyterian Hospital from 2004 to 2006, 10 were identified as being performed to evaluate known or suspected lymphoma and included a pre-splenectomy FDG-PET scan. The scans were assigned to low, intermediate or high splenic metabolic activity based on standardized uptake values (SUV). Low activity was associated with benign findings or mantle cell lymphoma at splenectomy, intermediate activity with marginal zone lymphoma and high activity with DLBCL. This comprises the largest pathologically confirmed series of cases to evaluate splenic FDG-PET uptake in suspected or known lymphoma. Low splenic SUV appears less likely to be associated with splenic involvement of lymphoma; intermediate and high values suggest presence of lymphoma. Our findings support a potential role for preoperative FDG-PET in consideration of the need for splenectomy in these settings. PMID- 18398740 TI - Pre-transplant 18FDG-PET predicts outcome in lymphoma patients treated with high dose sequential chemotherapy followed by autologous stem cell transplantation. AB - We evaluated the prognostic role of 18FDG-PET performed before ASCT in patients affected by lymphoma who underwent high-dose chemotherapy followed by ASCT as first-line treatment for high-risk disease or as second-line or more for relapsed or refractory disease. We retrospectively analyzed 53 consecutive patients, 14 with Hodgkin Lymphoma (HL) and 39 with non-Hodgkin Lymphoma (NHL), treated between February 1999 and October 2006 at our institution, who had a pre-ASCT FDG PET (pPET) evaluation. Median age was 45 years (range: 18-69). After a median follow-up of 31 months (range: 8-91), 7 out of 16 pPET+ patients and 10 out of 37 pPET- patients experienced lymphoma relapse. The 5-year OS is 90% and 55% (p = 0.01) in patients with negative and positive pPET, respectively. In conclusion, a positive pPET indicates a poorer outcome after ASCT with respect to a negative pPET; this subset of patients should be considered candidate to more intensive or investigational approaches. PMID- 18398741 TI - Favourable outcomes in children with diffuse large B-cell lymphoma treated by a short-term ALL-like regimen: a report on the NHL960 study from the Japanese Childhood Cancer and Leukemia Study Group. AB - In the NHL960 non-LB study, we treated diffuse large B-cell lymphoma (DLBCL) using a short-term ALL-like protocol. Thirty children up to 16 years of age with DLBCL were stratified into group 1 with stage I/II disease, or group 2 with stage III/IV disease. Their ages ranged from 9 months to 16 years of age, with a median of 9 years of age. The Murphy's stages were stage I in 7, stage II in 10, stage III in 6, and stage IV in 7 subjects. They received an ALL-like treatment without prophylactic cranial irradiation for 6 or 9 months. All children achieved a complete remission. Two patients with stage 3 disease experienced recurrences at 18 and 37 months after the start of chemotherapy. They responded to a short intensive regimen with Rituximab, followed by stem cell transplantation, and are alive without disease. The follow-up time ranged from 41 to 124 months with a median of 80 months. For all patients analyzed in this study, their overall survival and event-free survival (EFS) at 7-years was 100% and 93% +/- 4%, respectively. The 7-year EFS according to the treatment group was 100% for group 1, and 83% +/- 11% for group 2, respectively. PMID- 18398742 TI - Blastic mantle cell lymphoma with a Burkitt translocation. AB - MYC gene rearrangements are observed as a genetic change in the blastoid variant of mantle cell lymphoma (MCL). We present two patients, one had 30% atypical lymphocytes with Burkitt-like morphology in the peripheral blood and the other had 30% blasts in the bone marrow. Both had a CCND1/IGH and an MYC rearrangement by cytogenetics and FISH. The immunophenotype was CD5+[one was a weak positive], CD10+, CD20+, CCND1+, >90% Ki-67 proliferation fraction and CD23 negative. The diagnosis was blastic MCL with Burkitt features. These cases, along with the reported cases, distinguish an aggressive category of a blastic MCL with a poor prognosis that probably requires Burkitt therapy. PMID- 18398743 TI - Protein expression profiling of cytokines and cytokine receptors on purified chronic lymphocytic leukemia cells from patients with favourable prognostic indicators. AB - Chronic lymphocytic leukemia (CLL) cells rapidly undergo apoptosis when cultured in vitro, which contrasts with their prolonged survival in vivo. Multiple cytokines and cytokine receptors are believed to work together to regulate the survival of CLL cells. The aim of the current study was to measure the endogenous expression and secretion of cytokines and cytokine receptors in CLL cells when exogenous cytokines are minimized. We demonstrated that the intracellular and secreted levels of 174 cytokines and cytokine receptors of purified CLL B-cells were not significantly different from those of normal B-cells except for the secreted levels of IL-6 and eotaxin. IL-6 was 3.0 times lower (p = 0.038) whereas eotaxin was 2.2 times higher (p = 0.027) in CLL conditioned medium than in normal B-cell conditioned medium. Our results suggest that, except for IL-6 and eotaxin, CLL B-cells and their normal counterparts produce and secrete similar amounts of cytokines and cytokine receptors in vitro. PMID- 18398744 TI - Outcome of childhood B-cell non-Hodgkin lymphoma and B-cell acute lymphoblastic leukemia treated with the Tokyo Children's Cancer Study Group NHL B9604 protocol. AB - From June 1996 to January 2001, 91 patients with B-cell non-Hodgkin lymphoma or B cell acute lymphoblastic leukemia up to 18 years of age were enrolled in Tokyo Children's Cancer Study Group (TCCSG) NHL B9604 protocol study. Five-day intensive chemotherapy courses including high-dose methotrexate and high-dose cyclophosphamide were used for localized disease (Groups A and B). High-dose cytarabine was added for advanced disease (Groups C and D). Fifteen patients experienced an adverse event. There were three induction failures, eight relapses (three local, four bone marrow (BM), one BM + local), two toxic deaths and two second malignant neoplasm. Event-free survival at 6 years in Group D and in all patients was 82.4% +/- 9.2% and 81.9% +/- 4.4%, respectively. The TCCSG NHL B9604 protocol achieved an excellent treatment outcome especially in patients with the most advanced disease (Group D: high BM blast cell burden and/or central nervous system involvement). PMID- 18398745 TI - Mutated IgHV1-69 gene usage represents a distinct subgroup associated with indolent disease in chronic lymphocytic leukemia. AB - Biased IgHV gene usage in chronic lymphocytic leukemia (CLL) is well documented and suggests antigen involvement in leukemogenesis. IgHV1-69 is one of the most frequently rearranged IgHV genes in CLL and the majority of IgHV1-69 cases lack somatic hypermutation and display poor prognosis. However, its independent prognostic impact remains uncertain given reports showing a low proportion of mutated IgHV1-69 cases and stereotyped IgHV1-69 subsets with divergent clinical outcome. We assessed the frequency and clinical significance of IgHV1-69 gene usage in a cohort of 330 CLL patients. Functional IgHV1-69 gene rearrangements were detected in 32 cases (9.7%), 31 of which were characterised further. Seven (22.6%) were found to have undergone somatic hypermutation. This subgroup had shorter and more diverse complementarity determining region 3 (CDR3) sequences compared with unmutated IgHV1-69 cases. In addition, mutated IgHV1-69 gene status was associated with lower cell surface CD38 expression and less progressive disease as monitored by Binet staging, lymphocyte doubling time and requirement of chemotherapeutic intervention. To conclude, we present data confirming that IgHV1-69 gene rearrangements in CLL are not exclusively associated with unmutated IgHV status. In addition, we show that a somatically hypermutated subgroup may demonstrate more indolent characteristics despite the general association of IgHV1-69 gene usage with aggressive disease. PMID- 18398746 TI - Factors affecting autologous peripheral blood stem cell collection in patients with relapsed or refractory diffuse large cell lymphoma and Hodgkin lymphoma: a single institution result of 168 patients. AB - From 1996 to April 2006, 174 consecutive patients with relapsed or refractory diffuse large cell lymphoma (DLCL) and Hodgkin lymphoma (HL) received ESHAP as salvage and for mobilisation. Males 92, females 76. DLCL 64: HL 104, prior radiation in 35%. First relapse 45%, second relapse 12%, induction failure 43%. Median prior chemotherapy cycles were 6. Median age at apheresis was 26.5 years. Six patients failed mobilisation and 21 patients had CD34+ cells collection < 2 x 10(6)/kg on first apheresis. Median CD34+ cells/kg collection was 5.5 x 10(6)/kg for first apheresis and 6.7 x 10(6)/kg for all apheresis. We evaluated impact of histology, gender, age, stage, marrow involvement, prior radiation and chemotherapy cycles, timing (relapse1: relapse > 1: refractory), platelet count and weight. For first apheresis collection; all patients, younger age (p = 0.004), for DLCL (64), younger age (p = 0.021) and higher platelet count (p = 0.013) and for HL (104), younger age (p = 0.036) and male gender had better CD34+ cells collection. For all apheresis product, for all patients, age (p = 0.001) and no prior radiation therapy (p = 0.051) had better CD34+ cells collection. Higher first harvest CD34+ cells collection also resulted in early neutrophil (p < or = 0.001) and platelet (p = 0.004) engraftment. PMID- 18398747 TI - Cellular immune responses against the cancer-testis antigen SPAN-XB in healthy donors and patients with multiple myeloma. AB - The cancer-testis antigen SPAN-XB has been recently identified in multiple myeloma (MM). In the present study, we identified and characterized for the first time a cytotoxic cellular immune response against SPAN-XB in healthy donors and patients with MM. Using two independent computer algorithms, two SPAN-XB-derived peptides (peptides 624 and 626) with predicted binding to HLA-A2 were identified. To further improve the immunogenicity of peptide 626 we designed a heteroclitic peptide (peptide 627) by modifying one amino acid on the HLA binding position 2 of peptide 626. Using an IFN-gamma Elispot assay we could demonstrate the presence and functional activity of CD8 peptide specific T cells with all tested peptides. By analysis of peripheral blood of 13 healthy donors and five patients with MM peptide specific T-cell precursors specifically recognizing at least one of the tested peptides could be detected and expanded in 9 of 13 of tested donors and 3 of 5 tested patients. Importantly, in two donors specific peptides could be generated against the heteroclitic peptide 627 but not against the native peptide 626. We conclude that SPAN-XB-derived peptides can elicit a consistent CD8 T cell response in healthy donors and patients with MM. PMID- 18398748 TI - Anti proliferative activity of ELACY (CP-4055) in combination with cloretazine (VNP40101M), idarubicin, gemcitabine, irinotecan and topotecan in human leukemia and lymphoma cells. AB - This study evaluated combination drug partners for CP-4055, the C18:1(Delta9,trans) unsaturated fatty acid ester of cytarabine in HL-60 and U937 cells. Growth inhibition was assessed by ATP assay and drug interaction by the combination index and three dimensional methods. Synergy was observed in HL-60 cells for simultaneous combinations of CP-4055 with gemcitabine, irinotecan and topotecan, while combinations with cloretazine (VNP40101M) and idarubicin were additive. In U937 cells, synergy was observed with gemcitabine and additivity for the other drugs. In HL-60, the IC50 concentration of CP-4055 could be reduced 10 fold and that of gemcitabine 3-fold in combination versus the agents alone, an interaction that was independent of drug sequence, ratio and exposure time. In contrast, interactions of CP-4055 with the topoisomerase inhibitors became antagonistic when the drugs were administered 24 h prior to CP-4055 and at certain drug ratios, particularly in U937 cells. In summary, CP-4055 produced additive to synergistic anti proliferative activity when combined simultaneously with drugs from four mechanistic classes in cell culture models of human leukemia and lymphoma. The impact of drug sequence and ratio on the interactions argues for incorporation of these parameters into the design of combination chemotherapy regimens. PMID- 18398750 TI - Pharmacokinetic self-potentiation of idarubicin by induction of anthracycline carbonyl reducing enzymes. AB - Severe myelosuppression is one of the major adverse effects of Idarubicin (IDA). Especially, after two sequential therapy courses, IDA showed a stronger myelosuppression than after the first course. IDA was metabolized in liver by carbonyl reducing enzymes (CRE) to its 13-OH metabolite, idarubicinol(IDAol),which is more active compared with IDA. RLN-B2, a rat liver cell line, precultured in the presence of IDA showed higher CRE activity compared with non-precultured cells. A crude extract of the enzyme obtained from the livers of F344 rats preadministered IDA 7 days before they were sacrificed showed higher enzymatic activity than that from non-preadministered rats. At 4 h after IDA i.v., the production of IDAol was facilitated in the preadministered group compared with the non-preadministered group. In conclusion, CRE was induced by IDA pretreatment in vitro and vivo, resulting in increased IDAol, which could cause self-potentiation of the myelosuppressive and probably antitumor effects of IDA. PMID- 18398749 TI - Noxa mediates bortezomib induced apoptosis in both sensitive and intrinsically resistant mantle cell lymphoma cells and this effect is independent of constitutive activity of the AKT and NF-kappaB pathways. AB - Bortezomib is more active against mantle cell lymphoma (MCL) than against most other lymphoma subtypes. Nevertheless, up to half of patients with MCL have bortezomib resistant disease. Factors contributing to intrinsic resistance to bortezomib have not been determined. Here we used a panel of eight bortezomib sensitive (median IC(50) 5.9 nM) and three relatively bortezomib resistant cell lines (median IC(50) 12.9 nM) to investigate differences in tumor biology that could determine sensitivity to bortezomib. Bortezomib effectively inhibited high baseline proteasome activity and induced a comparable degree of proteasome inhibition in both sensitive and resistant cells. At 10 nM, bortezomib induced the proapoptotic BH3-only protein Noxa in sensitive but not resistant cells. At higher concentrations of bortezomib, however, Noxa was also upregulated in resistant cells and this effect was sufficient to induce apoptosis. Silencing of Noxa with siRNA rescued these cells from apoptosis, arguing against a defect in Noxa regulation or function as the basis of bortezomib resistance. Bortezomib was equally effective against cells with high and low constitutive NF-kappaB signaling. Also, sensitive and resistant MCL cell lines showed comparable activation of the AKT pathway. We conclude that bortezomib can overcome classic mechanisms of resistance to apoptosis and that determinants of bortezomib sensitivity in MCL are due to differences in signaling or stress pathways upstream of Noxa. PMID- 18398751 TI - Prolonged molecular remission in advanced phase of chronic myeloid leukemia using combination of imatinib and high-dose chemotherapy. PMID- 18398752 TI - Acute myeloid leukemia in Jehovah Witnesses. PMID- 18398753 TI - Simultaneous double mismatched cord blood transplantation in a young patient with secondary myelodysplastic syndrome: feasibility and complications. PMID- 18398754 TI - Clinical characteristics, prognostic factors and outcomes of Burkitt lymphoma in adult Asians. PMID- 18398755 TI - Biclonal T-cell receptor gammadelta+ large granular lymphocyte leukemia associated with rheumatoid arthritis. PMID- 18398757 TI - Not medical solutions, but overmedicalization by pharmaceutical company policies endanger both sexological care, science, and sexual medicine. A commentary. PMID- 18398758 TI - Psychodynamic issues raised by sexual medications: a prescriber's perspective. PMID- 18398759 TI - The DSM criteria of sexual dysfunction: need for a change. AB - This article discusses the flaws of the Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria of sexual dysfunctions. It focuses on issues such as what constitutes sexual dysfunction, whether the diagnostic criteria should include duration, intensity, and frequency, and whether distress should be used as a diagnostic criterion. Other controversial issues such as normalcy of sexual functioning and validity of some diagnostic entities are also discussed. The upcoming revision of the DSM diagnostic criteria should consider all these possible changes. Field trials to examine various criteria and their validity and reliability will probably be necessary. PMID- 18398760 TI - Do psychosexual factors play a role in the etiology of provoked vestibulodynia? A critical review. AB - The aim of this review was to critically examine published studies concerning the psychosexual aspects of provoked vestibulodynia. Despite the presence of several methodological limitations, some findings were consistently replicated. Overall, women with vestibulodynia demonstrate impaired sexual functioning, namely, lower levels of sexual desire, arousal, and frequency of intercourse. Childhood physical and sexual abuse represent potential risk factors for the development of this condition. Additionally, specific psychological states such as anxiety, fear of pain, hypervigilance, catastrophizing, and depression, are more frequently reported by these women. More rigorous studies are needed to establish which psychosexual variables may exacerbate and/or maintain vestibulodynia. PMID- 18398761 TI - Psychometric properties of the Psychotropic-Related Sexual Dysfunction Questionnaire (PRSexDQ-SALSEX) in patients with schizophrenia and other psychotic disorders. AB - Sexual dysfunction is a disturbing and often underrecognized problem associated with schizophrenia and its treatment. The Psychotropic-Related Sexual Dysfunction (PRSexDQ-SALSEX) is a brief and relatively nonintrusive questionnaire that has shown adequate psychometric properties in patients with depression. This study examined the psychometric properties of the PRSexDQ-SALSEX in a sample of patients with schizophrenia or other psychotic disorders who were experiencing anti-psychotic-induced sexual dysfunction and were switched to olanzapine. The PRSexDQ-SALSEX was very feasible and its internal reliability was satisfactory. In addition, this questionnaire showed a good convergent validity and sensitivity to tracking changes in sexual functioning. PMID- 18398762 TI - Sexual dysfunction as related to severity of fatigue in women with CFS. AB - To assess sexual function in women with chronic fatigue syndrome. The study included 27 women, aged 20 to 45 years, with chronic fatigue syndrome (CFS) and 15 healthy female controls. Sexual function was measured with the Golombok Rust Inventory of Sexual Satisfaction (GRISS) questionnaire and five clinical questions. In the patient group, total fatigue impact scale (FIS) score correlated with the GRISS satisfaction (r:-0.471, P < .005), avoidance (r: 0.632, P < .001) and sensuality (r: -0.445, P = .008) subscales. The GRISS satisfaction, avoidance, and sensuality subscale results and the fact of seeing the sexual act as a negative experience correlated with the intensity of fatigue in women with CFS. PMID- 18398763 TI - Infertility-related perceptions and responses and their associations with quality of life among rural chinese infertile couples. AB - An anonymous cross-sectional survey interviewed 192 infertile couples consulting a family planning clinic in rural China. Of them, over 30% believed that childless couples could not live well, 80% desired to have a child very badly, over 60% pressured themselves or spouse due to infertility, and over 50% felt pressured when having sex. Furthermore, 19.8% of men and 37.5% of women felt that infertility is humiliating for women. A multivariate analyses showed that a lower income, a worsened spousal relationship, infertility related perceptions, pressuring oneself or spouse due to infertility, and a strong desire for children were significantly associated with a lowered quality of life. Gender differences were also observed. PMID- 18398764 TI - The unconsummated marriage: its frequency and clinical characteristics in a sexual dysfunction clinic. AB - The object of this study was to investigate frequency of the unconsummated marriages and its clinical characteristics among the subjects who referred to a sexual dysfunction clinic in Turkey. Four hundred and forty-nine unconsummated marriage cases were evaluated among 1880 subjects referred to Acibadem Sexual Health Clinic, Istanbul, between December 2000 and December 2004. Unconsummated marriage rose from the female partner in 67% of the cases, 7% males, and 26% both. Vaginismus was the most prominent cause in 81% of the cases, erectile dysfunction in 10.5%, premature ejaculation in 5%. Previous reports from Eastern societies including Turkey indicate higher incidences of premature ejaculation and vaginismus than the Western world which underline a strong cultural influence in the background of these disorders. These also appeared to be the most important precursors of unconsummated marriage in our study. PMID- 18398766 TI - Inferring species membership using DNA sequences with back-propagation neural networks. AB - DNA barcoding as a method for species identification is rapidly increasing in popularity. However, there are still relatively few rigorous methodological tests of DNA barcoding. Current distance-based methods are frequently criticized for treating the nearest neighbor as the closest relative via a raw similarity score, lacking an objective set of criteria to delineate taxa, or for being incongruent with classical character-based taxonomy. Here, we propose an artificial intelligence-based approach - inferring species membership via DNA barcoding with back-propagation neural networks (named BP-based species identification) - as a new advance to the spectrum of available methods. We demonstrate the value of this approach with simulated data sets representing different levels of sequence variation under coalescent simulations with various evolutionary models, as well as with two empirical data sets of COI sequences from East Asian ground beetles (Carabidae) and Costa Rican skipper butterflies. With a 630-to 690-bp fragment of the COI gene, we identified 97.50% of 80 unknown sequences of ground beetles, 95.63%, 96.10%, and 100% of 275, 205, and 9 unknown sequences of the neotropical skipper butterfly to their correct species, respectively. Our simulation studies indicate that the success rates of species identification depend on the divergence of sequences, the length of sequences, and the number of reference sequences. Particularly in cases involving incomplete lineage sorting, this new BP-based method appears to be superior to commonly used methods for DNA-based species identification. PMID- 18398767 TI - Testing the reliability of genetic methods of species identification via simulation. AB - Although genetic methods of species identification, especially DNA barcoding, are strongly debated, tests of these methods have been restricted to a few empirical cases for pragmatic reasons. Here we use simulation to test the performance of methods based on sequence comparison (BLAST and genetic distance) and tree topology over a wide range of evolutionary scenarios. Sequences were simulated on a range of gene trees spanning almost three orders of magnitude in tree depth and in coalescent depth; that is, deep or shallow trees with deep or shallow coalescences. When the query's conspecific sequences were included in the reference alignment, the rate of positive identification was related to the degree to which different species were genetically differentiated. The BLAST, distance, and liberal tree-based methods returned higher rates of correct identification than did the strict tree-based requirement that the query was within, but not sister to, a single-species clade. Under this more conservative approach, ambiguous outcomes occurred in inverse proportion to the number of reference sequences per species. When the query's conspecific sequences were not in the reference alignment, only the strict tree-based approach was relatively immune to making false-positive identifications. Thresholds affected the rates at which false-positive identifications were made when the query's species was unrepresented in the reference alignment but did not otherwise influence outcomes. A conservative approach using the strict tree-based method should be used initially in large-scale identification systems, with effort made to maximize sequence sampling within species. Once the genetic variation within a taxonomic group is well characterized and the taxonomy resolved, then the choice of method used should be dictated by considerations of computational efficiency. The requirement for extensive genetic sampling may render these techniques inappropriate in some circumstances. PMID- 18398768 TI - Genomic outposts serve the phylogenomic pioneers: designing novel nuclear markers for genomic DNA extractions of lepidoptera. AB - Increasing the number of characters used in phylogenetic studies is the next crucial step towards generating robust and stable phylogenetic hypotheses - i.e., strongly supported and consistent across reconstruction method. Here we describe a genomic approach to finding new protein-coding genes for systematics in nonmodel taxa, which can be PCR amplified from standard, slightly degraded genomic DNA extracts. We test this approach on Lepidoptera, searching the draft genomic sequence of the silk moth Bombyx mori, for exons > 500 bp in length, removing annotated gene families, and compared remaining exons with butterfly EST databases to identify conserved regions for primer design. These primers were tested on a set of 65 taxa primarily in the butterfly family Nymphalidae. We were able to identify and amplify six previously unused gene regions (Arginine Kinase, GAPDH, IDH, MDH, RpS2, and RpS5) and two rarely used gene regions (CAD and DDC) that when added to the three traditional gene regions (COI, EF-1alpha and wingless) gave a data set of 8114 bp. Phylogenetic robustness and stability increased with increasing numbers of genes. Smaller taxanomic subsets were also robust when using the full gene data set. The full 11-gene data set was robust and stable across reconstruction methods, recovering the major lineages and strongly supporting relationships within them. Our methods and insights should be applicable to taxonomic groups having a single genomic reference species and several EST databases from taxa that diverged less than 100 million years ago. PMID- 18398769 TI - Maximum likelihood supertrees. AB - We analyze a maximum likelihood approach for combining phylogenetic trees into a larger "supertree." This is based on a simple exponential model of phylogenetic error, which ensures that ML supertrees have a simple combinatorial description (as a median tree, minimizing a weighted sum of distances to the input trees). We show that this approach to ML supertree reconstruction is statistically consistent (it converges on the true species supertree as more input trees are combined), in contrast to the widely used MRP method, which we show can be statistically inconsistent under the exponential error model. We also show that this statistical consistency extends to an ML approach for constructing species supertrees from gene trees. In this setting, incomplete lineage sorting (due to coalescence rates of homologous genes being lower than speciation rates) has been shown to lead to gene trees that are frequently different from species trees, and this can confound efforts to reconstruct the species phylogeny correctly. PMID- 18398771 TI - Frequent detection of benzodiazepines in drugged drivers in Norway. AB - OBJECTIVE: To describe the Norwegian system for handling suspected drugged driving cases according to an impairment-based law, with primary focus on benzodiazepines (BZDs), blood concentrations and combination with other psychoactive compounds. METHODS: Routines for handling suspected driving under the influence of drugs other than alcohol are described. These include primary police investigation, blood sampling, and clinical tests of impairment performed by a police physician, a standard analytical program covering the most relevant illegal drugs and medicines relevant to traffic safety (approximately 25 compounds), and expert witness statements prepared for the court. The drug use patterns, blood drug concentrations, and frequency of multi-drug use have been recorded, with primary focus on benzodiazepines (BZDs). Use of BZDs among apprehended drivers has been compared with patient prescriptions recorded for the same BZDs. RESULTS: One or more drugs have been detected in approximately 80% of the cases received for analysis every year. BZDs have been the most prevalent drugs and have been detected in 38-57% of the cases, which is more frequent than other common illegal drugs; e.g., tetrahydrocannabinol (THC; 30-43%) and amphetamine (33-39%). The majority of the BZDs have been detected at supratherapeutic blood concentrations and frequently in combination with illegal drugs, other psychoactive medicines, or alcohol. Less than 5% of the BZDs (except for nitrazepam - 7.6%) have been found to be the only drug present at therapeutic blood levels. The majority of the drivers were 20-39 years old (median age 29 33), while the majority of BZDs prescribed were to users over 50 years of age. CONCLUSIONS: Drivers with BZD detected are probably not representative of ordinary patients with BZD prescriptions, as shown by the age disparity of drivers and patients. The frequent detection of BZDs suggests that these compounds should be included in the analytical program used for blood samples from apprehended drivers and for studies on drug involvement in road traffic accidents and risk calculations. PMID- 18398770 TI - Licensing teenagers: nontraffic risks and benefits in the transition to driving status. AB - OBJECTIVE: The initiation of teen driving, which occurs between ages 15 and 17, has generally been studied primarily in relation to crash injury reduction. However, it may be the most important period influencing development between puberty and emerging adulthood because, once the teens are driving on their own or riding with other teens, parental control over social behavior is greatly reduced and opportunities for risk-taking are substantially increased. The noncrash risks and benefits of licensing during this critical transition period are the subject of this paper. METHODS: We identify "transition teens" as a subgroup of all 15-to 17-year-olds who reside in middle-class suburban and rural areas where families can afford a car and where a vehicle is needed for reasonable mobility. In this paper, we review recent studies on the effect of driving on non-traffic-related behaviors of teen drivers. Our analysis suggests that there are two environments that affect behavior and development: the home environment and the extended environment. Before driving age, the home environment influences are omnipresent in the life of the developing child. However, once teens can move more readily within the community and travel outside the home environment on their own or with a peer, they have access to locations where many or all the local controls on risky behavior may be absent. RESULTS AND CONCLUSIONS: This article introduces the concept of "transitional teens" and provides a rough model that explains the possible risks encountered by this group when they begin to drive a vehicle or to ride with a peer. We offer this model to the research community with a challenge to hypothesize, test, and explore this proposed transitional period so that we can better understand and mitigate its associated risks. PMID- 18398772 TI - The relation between the use of psychoactive substances and the severity of the injury in a group of crash-involved drivers admitted to a regional trauma center. AB - OBJECTIVE: There is much evidence that driving under the influence of alcohol and/or drugs of abuse is related to an increased accident risk. A remaining question is whether the use of psychoactive substances is also related to clinically more severe accidents. The aim of this study is to explore the relationship between the use of psychoactive substances and the injury severity in a group of crash-involved drivers. METHODS: The study group included all injured car drivers, admitted to the regional trauma center, in the period from May 2000 until August 2001. The outcome of interest was the severity of injury, measured by using the Injury Severity Score (ISS). The determinant was the presence of psychoactive substances in blood and urine samples. Psychoactive substances tested for were alcohol, amphetamines, barbiturates, benzodiazepines, cannabis, methadone, opiates, and tricyclic antidepressants in blood and urine. RESULTS: The number of injured car drivers included in this study was 106. Overall, 43% (46/106) of the drivers tested positive for at least one psychoactive substance. Comparison of the means of the log ISS suggests that there is no significant difference between drivers who tested positive for alcohol and/or drugs, compared to drivers tested negative. CONCLUSION: The results of this study support the hypothesis that there is no clear association between use of psychoactive substances and the severity of crash-related injury. PMID- 18398773 TI - DUI offenders who delay relicensing: a quantitative and qualitative investigation. AB - OBJECTIVES: As in many jurisdictions, individuals convicted of driving under the influence (DUI) in the province of Quebec are mandated to relicensing programs, which include obligatory participation in intervention programs. However, prolonged delay in relicensing is widespread, potentially contributing to unlicensed driving, untreated substance misuse problems, and drink-driving risk. Information about the characteristics of DUI offenders who delay relicensing (DR) is sparse. This investigation compares the characteristics of DR offenders with those offenders who do not delay (NoDR). In addition, the rationales of DR offenders for delaying relicensing are explored qualitatively. METHODS: Two studies were conducted to explore the characteristics of DR offenders. In Study 1, DR offenders (n = 46) were compared to NoDR offenders (n = 74) on multidimensional measures of psychosocial functioning, driving behavior, substance use, and psychological and neurocognitive characteristics. In Study 2, a qualitative examination of 20 DR offenders' reasons underlying delayed relicensing was undertaken, with verbatims content analyzed to identify major themes. A questionnaire, based upon this preliminary analysis, was then administered to another sample of DR participants (N = 37) to appraise and confirm thematic comprehensiveness. RESULTS: The main findings of Study 1 were that, compared to NoDR offenders, DR offenders had more past DUI convictions, were at greater risk for drink driving per kilometer (km) driven, were more likely to have received substance abuse treatment, and exhibited indices of poorer neurocognitive performance in visual memory and behavioral inhibition domains. No group differences were uncovered on substance use measures. The findings of Study 2 revealed that the expense of participation, availability of alternate transportation, lack of interest, and no access to a vehicle were the most frequent explanations for delayed relicensing. CONCLUSIONS: Overall, these findings suggest that both individual and contextual factors influence timely fulfillment of relicensing requirements. While the cost of relicensing may succeed in removing some offenders from the road, it may also be a barrier for others at risk for drink driving, preventing exposure to needed intervention programs. Reducing this barrier may need to be weighted against the risks of relicensing more DUI offenders. Neurocognitive factors may need to be taken into account to not only decrease delay in relicensing but also increase the benefits from participation in interventions that are part of current relicensing programs. PMID- 18398774 TI - Traffic speeds on interstates and freeways 10 years after repeal of national maximum speed limit. AB - OBJECTIVES: Essentially all published analyses of operational and safety outcomes related to enactment or repeal of the national maximum speed limit (NMSL) were based on data limited to the initial 1-3 years. The purpose of the present study was to collect and analyze longer term speed data. METHOD: In 2006, traffic speeds were surveyed at 26 locations on urban and rural expressways in five states (California, Montana, Nevada, New Mexico, and Texas) where speeds had been measured in 1996, immediately after repeal of NMSL. Most speed limits were unchanged during the approximately 10-year period. However, Montana introduced a numeric 75 mph limit for passenger vehicles in place of a "reasonable and prudent" limit. In Texas, urban freeway speed limits for passenger vehicles were reduced 5 mph, and truck limits were increased 5 mph. RESULTS: On rural interstates without speed limit changes, travel speeds increased for both passenger vehicles and large trucks; the proportion of passenger vehicles exceeding 80 mph tripled. On rural interstates in Montana where speed limits were lowered for passenger vehicles, travel speeds decreased, even for large trucks whose speed limits had not changed. On urban freeways where speed limits did not change, travel speeds declined somewhat for both passenger vehicles and large trucks; during the study period there also were large increases in traffic volume and development of surrounding areas. On urban freeways in Texas where speed limits declined for passenger vehicles, travel speeds generally decreased, even for large trucks whose speed limits actually had increased. CONCLUSIONS: The data suggest that where traffic volumes allow, travel speeds still are increasing 10 years after repeal of NMSL, and many drivers are speeding. The study also shows that speed increases can be curbed and even reversed when speed limits are lowered. Increased enforcement is needed to curb speed increases and the increased risk of serious crashes. PMID- 18398775 TI - Motorcycle helmet use in Southern China: an observational study. AB - OBJECTIVE: In China, despite national motorcycle helmet legislation and the known safety benefits of helmets, helmet use remains low. The aim of this study was to determine the rate of motorcycle helmet use and attitudes towards helmet use among drivers and passengers in two cities in Southern China to provide baseline data and scientific evidence for the formulation of an intervention aimed at strengthening road safety law enforcement. METHODS: Observational sites were randomly selected from three road types (national roads, main streets, and subordinate streets). Observations were conducted during six specified time periods and uniform checklists were used to record helmet use. Motorcycle riders were randomly selected from service stations, elementary schools, and supermarket car parks to participate in a face-to-face interview to ascertain attitudes. RESULTS: Overall, the rate of correct helmet use among drivers was higher in Chaozhou (34.6%) than in Shantou (30.2%; P < 0.001). Correct helmet use was higher among drivers in main streets, during daytime hours, and during weekdays (P < 0.001). The level of helmet knowledge of motorcycle drivers was high with most reporting that helmets prevent or reduce head injury (Shantou: 78.2%; Chaozhou 70.6%). CONCLUSIONS: Although level of awareness of the benefits of helmets was high, observed helmet usage was low. These results suggest that there is a need to implement new interventions to increase helmet use. PMID- 18398776 TI - Parents' knowledge and attitudes about preventing injuries in motor vehicle accidents in children in Greece. AB - OBJECTIVE: The aim of this study was to record parents' knowledge and perceptions about the adoption of the appropriate preventive measures in order to avoid serious injury or death of children (as passengers, pedestrians, or bicyclists) in case of motor vehicle accidents. METHODS: A total of 1000 parents of children, aged 0-14 years old, served as the basis for this study; 854 were parents of students in kindergartens, elementary schools and high schools from both public and private sectors, and 146 were parents of children who visited the outpatient clinic of the Paediatric Department of three major hospitals in Athens, Greece, for various reasons, except for accidents, from Sept. 1, 2004, to Aug. 31, 2005. All participants answered an anonymous questionnaire structured for the needs of this study. RESULTS: Over forty-one percent (41.4%) of the participants believed that they were not sufficiently informed about preventing childhood injuries in car accidents, while 80.4% insisted that children under 12 years old comprehended the concepts of collision, distance, speed, and signals of traffic policemen. Parents' information came from the media (65.6%), brochures (43%), posters (9.4%), and health care professionals (14.2%). Child seats, seat belts, and helmets were not used uniformly (76.1%, 61.8%, and 7.4%, respectively), while the majority of parents (88.4%) positioned their children in the back seat of the vehicle. The results arising from the statistical analysis (p = 0.05) showed that parents' information about preventing childhood injuries in car accidents was associated with mother's age (p = 0.07), father's (p = 0.042) and mother's educational level (p = 0.003), as well as with father's profession (p = 0.031). An association was also found between parents' information about preventing injuries in car accidents and children having been in an accident in the past as passengers (p = 0.042) or bicyclists (p = 0.011). CONCLUSIONS: The adoption of harsh measures for preventing the devastating consequences of car accidents in children is of high importance. Parents' rigorous education must become our priority. Health care professionals need to act dynamically in sensitizing Greek parents. PMID- 18398777 TI - Motorcycle helmets in Vietnam: ownership, quality, purchase price, and affordability. AB - OBJECTIVES: This study investigated motorcycle helmet ownership, quality, purchase price, and affordability in Vietnam. METHOD: A random sample of motorcyclists was interviewed to investigate aspects of helmet ownership, the purchase price, and affordability of a motorcycle helmet. Multivariate modeling conducted to determine factors associated with the purchase price and affordability of motorcycle helmets. Helmet quality was assessed based on current legal requirements in Vietnam. RESULTS: The prevalence of helmet use in Vietnam remains low (23.3%) despite a high level of helmet ownership (94%), indicating that this is an important area for public health intervention. Overall the quality of helmets appeared to be good; however, few helmets displayed legally required information. Motorcyclists with a high income purchase more helmets for their household rather than more expensive helmets. CONCLUSION: To ensure that helmets are accessible to the community, policy-makers need to consider pricing motorcycle helmets at a price indicated by the results of this study. Prior to universal motorcycle helmet legislation, the government will also need to ensure that standard helmets are available and that enforcement is at a level to ensure that motorcycle helmets are actually used. PMID- 18398778 TI - Analysis of head impacts causing neck compression injury. AB - OBJECTIVE: Human cadavers have been subjected to inverted drop, linear, and pendulum impacts to the top of the head, causing neck compression injury. The data are not comparable on the basis of impact velocity because of differing impact masses and test conditions. This study analyzed the published biomechanical data and used peak head velocity to merge the datasets. Correlations were determined between biomechanical responses and serious injury (AIS 3+). METHODS: Three studies were found involving 33 inverted drop tests and three others involving 42 linear or pendulum impacts to the top of a cadaver's head. Various biomechanical responses were measured in the tests. The datasets could not be meaningfully merged on the basis of impact velocity. The coefficient of restitution (e) was determined and the peak head velocity calculated for tests with missing data. This allowed the datasets to be merged and statistically analyzed for relationships between head velocity, impact force, and serious injury. Power functions were fit to the biomechanical data, t-tests conducted for significant differences in injury, and logit risk functions determined. RESULTS: The coefficient of restitution was e = 0.24 +/- 0.16 (n = 19) for the drop tests and e = 0.21 +/- 0.12 (n = 20) for the impact tests. Peak head velocity was 22% higher than the impact velocity for the drop tests but -20% lower in the impact tests. Head velocity averaged 6.32 +/- 1.29 m/s (n = 51) causing serious injury and 3.75 +/- 2.16 m/s (n = 24) without injury (t = 5.39, p = 0.00001, df = 31). Impact force was 7,382 +/- 3,632 N with injury and 3,760 +/- 3,528 N without (t = 3.95, p = 0.0003, df = 42). A power function fit the impact force versus head velocity data (F = 374V(h)(1.565), R(2) = 0.758). CONCLUSION: Peak head velocity was determined for inverted drop and impact tests as a means of merging and analyzing cadaver data on serious injury for impacts to the top of the head. There are relationships between head velocity, impact force, and serious injury. A 15% risk of serious injury is at 2.3 m/s (5.1 mph) head velocity and 50% risk at 4.2 m/s (9.4 mph); however, more data are needed in the 2-4 m/s head velocity range to clarify injury risks. In addition, many factors influence the risk of the neck injury, including the age and physical condition of the person; orientation of the head, neck, and torso; and location of impact and interface. PMID- 18398779 TI - Traumatic rupture of thoracic aorta in real-world motor vehicle crashes. AB - OBJECTIVE: Traumatic rupture of thoracic aorta (TRA) was reported in the literature to be a major cause of death in motor vehicle crashes. This study aims at evaluating the most relevant risk factors of TRA. It also aims at analyzing the types of TRA as a function of car crash conditions and rib cage fractures. METHODS: In-depth crash data collected from 1998 to 2006 as part of the Co operative Crash Injury Study (CCIS) were retrospectively investigated to assess frontal, near-side, and far-side injury risks. This database includes 15,074 occupants with individual detailed autopsy reports. Multivariate statistical analyses were performed. The influence of the following variables on TRA occurrence was studied: gender and age of the involved occupant, ETS, compartment intrusion, and restraint system. Features of TRA and rib cage fractures were described thanks to autopsy data. RESULTS: Although TRA occurred in only 1.2% of all occupants, TRA victims accounted for 21.4% of all fatalities. The incidence of TRA was found twice higher in side impacts (2.4%) than in frontal ones (1.1%). TRA injury risk increased with ETS, intrusion, and age and decreased with the absence of intrusion regardless of the impact direction. It also decreased for belted occupants in frontal impacts. Except for the site of injury, the TRA features were similar whatever the crash conditions. The multiple ribs fractures were the most common injuries associated with TRA (79.1%) and TRA victims with uninjured or slightly injured (AIS 1) rib cage were significantly younger (p < 0.0001) than other TRA victims. Whatever the impact type, the TRA victims sustained mostly bilateral rib fractures (68%). Results also emphasized a close relationship between the principal direction of force and the body side with more fractured ribs. However, whatever the impact type, the aortic injury site or the side of the thorax, fractures concerned mainly the 2nd up to the 7th ribs of TRA victims. CONCLUSIONS: This study emphasized four main variables influencing the TRA occurrence: ETS, compartment intrusion, age, and seat belt use. The results suggested that the injury site may be different depending on the occupant or the impact type. However, the typical TRA, i.e., a partial or complete aorta transection within the peri-isthmic region, affected any occupant independently of age and impact type. The high frequency of bilateral rib cage fractures observed in TRA victims and the significant influence of intrusion on TRA occurrence emphasized that the aortic injury mechanism mainly involves a severe direct chest impact or compression. PMID- 18398780 TI - Chest deflections and injuries in oblique lateral impacts. AB - A majority of laboratory-driven side-impact injury assessments are conducted using postmortem human subjects (PMHS) under the pure lateral mode. Because real world injuries occur under pure and oblique modes, this study was designed to determine chest deflections and injuries using PMHS under the latter mode. Anthropometrical data were obtained and x-rays were taken. Specimens were seated on a sled and lateral impact acceleration corresponding to a change in velocity of 24 km/h was applied such that the vector was at an angle of 20 or 30 degrees. Chestbands were fixed at the level of the axilla (upper), xyphoid process (middle), and tenth rib (lower) location. Deflection contours as a function of time at the levels of the axilla and mid-sternum, representing the thorax, and at the tenth rib level, representing the abdomen, were evaluated for peak magnitudes. All data were normalized using mass-scaling procedures. Injuries were identified following the test at autopsy. Trauma graded according to the Abbreviated Injury Score, 1990 version, indicated primarily unilateral rib fractures and soft tissue abnormalities such as lung contusion and diaphragm laceration occurred. Mean peak deflections at the upper, middle, and lower levels of the chest for the 30-degree tests were 96.2, 78.5, and 76.8 mm. For the 20 degree tests, these magnitudes were 77.5, 89.9, and 73.6 mm. Statistical analysis indicated no significant (p > 0.05) differences in peak chest deflections at all levels between the two obliquities although the metric was significantly greater in oblique than pure lateral impacts at the mid and lower thoracic levels. These response data are valuable in oblique lateral impact assessments. PMID- 18398781 TI - Patients treated with obstructive sleep apnea syndrome and fitness to drive assessment in clinical practice in Spain at the medical traffic centers. AB - OBJECTIVE: To assess fitness to drive among drivers with treated obstructive sleep apnea syndrome. METHODS: A total of 5234 drivers who attended two Spanish Medical Driver Test Centers for assessment of their fitness to drive prior to renewing or issuing their driving license were included in the study. Information regarding sociodemographic aspects, patterns of driving, medical condition, medication use, and patterns of alcohol consumption was recorded. RESULTS: Eleven out of 5234 drivers (0.2%) suffered from obstructive sleep apnea syndrome, all of them being treated with continuous positive airway pressure, and they were found fit to drive with restrictions. CONCLUSIONS: A lower than expected prevalence of patients with obstructive sleep apnea syndrome was found among drivers undergoing fitness to drive assessment. Suffering from obstructive sleep apnea syndrome does not prevent driving as long as the patient is undergoing adequately controlled treatment. PMID- 18398782 TI - Analyses of factors of crash avoidance maneuvers using the general estimates system. AB - OBJECTIVE: Taking an effective corrective action to a critical traffic situation provides drivers an opportunity to avoid crash occurrence and minimize crash severity. The objective of this study is to investigate the relationship between the probability of taking corrective actions and the characteristics of drivers, vehicles, and driving environments. METHODS: Using the 2004 GES crash database, this study classified drivers who encountered critical traffic events (identified as P_CRASH3 in the GES database) into two pre-crash groups: corrective avoidance actions group and no corrective avoidance actions group. Single and multiple logistic regression analyses were performed to identify potential traffic factors associated with the probability of drivers taking corrective actions. RESULTS: The regression results showed that the driver/vehicle factors associated with the probability of taking corrective actions include: driver age, gender, alcohol use, drug use, physical impairments, distraction, sight obstruction, and vehicle type. In particular, older drivers, female drivers, drug/alcohol use, physical impairment, distraction, or poor visibility may increase the probability of failing to attempt to avoid crashes. Moreover, drivers of larger size vehicles are 42.5% more likely to take corrective avoidance actions than passenger car drivers. On the other hand, the significant environmental factors correlated with the drivers' crash avoidance maneuver include: highway type, number of lanes, divided/undivided highway, speed limit, highway alignment, highway profile, weather condition, and surface condition. Some adverse highway environmental factors, such as horizontal curves, vertical curves, worse weather conditions, and slippery road surface conditions are correlated with a higher probability of crash avoidance maneuvers. These results may seem counterintuitive but they can be explained by the fact that motorists may be more likely to drive cautiously in those adverse driving environments. CONCLUSIONS: The analyses revealed that drivers' distraction could be the highest risk factor leading to the failure of attempting to avoid crashes. Further analyses entailing distraction causes (e.g., cellular phone use) and their possible countermeasures need to be conducted. The age and gender factors are overrepresented in the "no avoidance maneuver." A possible solution could involve the integration of a new function in the current ITS technologies. A personalized system, which could be related to the expected type of maneuver for a driver with certain characteristics, would assist different drivers with different characteristics to avoid crashes. Further crash database studies are recommended to investigate the association of drivers' emergency maneuvers such as braking, steering, or their combination with crash severity. PMID- 18398783 TI - Endoscopic surveillance of the ileoanal pouch following restorative proctocolectomy for familial adenomatous polyposis. AB - Persistence of underlying disease in the residual rectal mucosa and anal transition zone occurs following mucosectomy with either a hand-sewn anastomosis or a double-stapled anastomosis. Furthermore, recent reports have suggested an increased incidence of neoplasia in the pouch body. For this reason, endoscopic surveillance is performed not only as a screening tool to detect significant intraepithelial neoplastic lesions but also with secondary therapeutic intent aimed at reducing the adenoma burden within the ileoanal pouch. Conventional endoscopic assessment of the ileoanal pouch can be challenging. In the future, novel adjunctive endoscopic technologies such as magnification endoscopy and confocal endomicroscopy may improve our diagnostic and therapeutic management of this group. PMID- 18398784 TI - Estrogen receptors in human pituitary tumors. AB - The relationship between the presence of estrogen receptors in pituitary adenomas and the post surgical evolution of the patients in order to find another prognostic parameter for these tumors have been studied to improve the treatment selection. Estrogen receptors were studied by immunocytochemistry in histological sections of paraffin embedded 42 pituitary adenomas. Only 19% of these tumors were positive for estrogen receptors. As expected, the higher concentration (60%) was found in prolactin secreting adenomas, although we found estrogen receptors in one somatotroph and in one nonsecreting. The most aggressive tumors were those negative for estrogen receptors within the prolactinomas and the positive somatotrophinoma. The results of this work suggest that the determination of estrogen receptors in pituitary tumors might help as a prognostic factor in these adenomas. PMID- 18398785 TI - [Pathophysiology of the obesity hypoventilation syndrome]. AB - The obesity hypoventilation syndrome (OHS) is defined by extreme overweight (BMI 30 kg/m2), daytime hypoventilation (PaCO2 > 45 mm Hg, the absence of other known causes of hypoventilation) and sleep-related breathing disorders. Obesity impairs breathing due to a restrictive ventilatory disorder, reduction of the capacity of respiratory muscles and diminishment of the ventilatory response. The restriction cannot serve as the only explanation of OHS because body weight or compliance on the one hand and hypoventilation on the other hand only correlate weakly. Obesity increases the work of breathing by greater body mass with its increased oxygen demand, impaired diaphragmatic mobility, upper airway obstruction, and oxygen desaturation which result in an inadequacy of oxygen demand and supply. The adjustment of the chemoreceptors can avoid the overload on the capacity of the respiratory muscles, at least in a number of patients or in the course of the disease. This disproportion results in hypercapnia. Furthermore, the level of leptin is an important factor in the pathophysiology of OHS. The blood level of leptin correlates with the body fat mass in humans. However, there seems to be a relative leptin deficiency in the brain in overweight humans. Therefore, in contrast to animals, leptin cannot sufficiently increase ventilation in man to avoid hypercapnia. PMID- 18398786 TI - [Clinical features and diagnosis of bronchopulmonary infections in the elderly]. AB - Diseases of the lung are one of the main causes of morbidity and mortality in the elderly. The risk of respiratory infections is increased due to structural changes, malnutrition, co-morbidity, and a variety of other factors. Bacterial and viral pathogens cause acute bronchitis and exacerbations of chronic bronchitis (AECB). Community acquired pneumonias (CAP) show a different spectrum of pathogens and clinical course in comparison to nosocomial pneumonias (hospital acquired pneumonia, HAP). Institutionalised patients are at risk of a health care associated pneumonia (HCAP), with often a different spectrum of pathogens in comparison to CAP and HAP. Elderly patients with cerebrovascular disease and impairment of swallowing or cough reflexes often suffer from aspiration pneumonias. The mortality is highest in the elderly, comorbid, and immunocompromised patient with nosocomial pneumonia. Important preventive measures include influenza and pneumococcal vaccination, avoidance of immobility, oral hygiene, and sufficient nutrition. PMID- 18398787 TI - [Friend or foe: combination therapy with inhaled corticosteroids and long-acting beta2-agonists in chronic obstructive pulmonary disease (COPD)]. AB - Inhaled corticosteroids (ICS) used in COPD (chronic obstructive pulmonary disease) are recommended only in combination with a long-acting beta2-agonist (LABA) in stage 3 and higher in COPD treatment guidelines. In comparison to placebo and the single components, a superior control by means of the ICS/LABA fixed combination therapy has been demonstrated for clinical improvement in the following parameters: reduction of exacerbation rate and hospitalisations, reduction of dyspnoea and improvement of forced expiratory volume in one second (FEV1). In contrast to data from database studies, the large prospective TORCH (Towards a Revolution in COPD Health) trial found in the ICS/LABA group a beneficial effect on the reduction of mortality only as a trend in the ICS/LABA group, which did not reach statistical significance. In long-term trials, ICS treated patients experienced up to 10% oral and/or pharyngeal candidiasis. ICS was associated with an excess risk of pneumonia, which doubles the pneumonia incidence in patients not receiving ICS. The probability of having pneumonia reported as an adverse event was 18-19 % in the ICS groups and resulted in a 1.7 2.2 elevated pneumonia risk. Because ICS therapy is recommended only in conjunction with a bronchodilator, fixed ICS/LABA combinations are a logical consequence for COPD long-term therapy. PMID- 18398788 TI - [Clinical features and diagnosis of tuberculosis]. AB - Recently, major advances have been accomplished in the diagnosis of active tuberculosis. A comprehensive diagnostic approach for a patient with possible tuberculosis includes a detailed medical history and clinical examination as well as the results of radiological, microbiological, immunological, molecular biological and histological methods. In concert, these results enable the clinician to rapidly develop a decision with a high probability for the diagnosis or exclusion of active tuberculosis. Therapeutic intervention can thus be made early, even though corrections in these decisions need to be considered depending on the results of Mycobacterium tuberculosis culture and sensitivity testing. PMID- 18398789 TI - [Influence of statins on the antiplatelet effect of clopidogrel and on cardiovascular outcome in patients after coronary intervention]. AB - BACKGROUND AND OBJECTIVE: Statins are frequently given in conjunction with clopidogrel for prophylaxis and therapy of coronary heart disease. The antiplatelet effect of clopidogrel may be attenuated by lipophilic statins such as atorvastatin. It was the aim of this study to measure the antiplatelet effect of clopidogrel and the incidence of cardiovascular events after stent implantation in patients with coronary artery stenting. PATIENTS AND METHODS: ADP induced aggregometry was used in 319 patients after coronary artery stent implantation to determine whether added administration of clinically relevant statins would affect the antiplatelet action of clopidogrel. Also a three-month follow-up was undertaken to determine the incidence of cardiovascular events. RESULTS: There were no significant differences between the statin-receiving subgroups, either in platelet aggregation nor in clinical outcome in our cohort. CONCLUSION: No statin-clopidogrel interaction nor any clinically relevant events were documented in the studied cohort. No interaction between CYP3A4 statins and a single loading dose of 600 mg clopidogrel was documented in this cohort. PMID- 18398790 TI - [Electrocardiographic changes in patients presenting with an acute coronary syndrome: "apical ballooning" versus anterior myocardial infarction]. AB - BACKGROUND AND OBJECTIVE: Transient left ventricular (LV) apical ballooning (AB) is characterized by chest pain, electrocardiographic (ECG) changes and LV apical akinesia mimicking anterior myocardial infarction (AMI) in the absence of coronary artery disease. This study aimed to assess if the ECG could reliably differentiate between these two syndromes with a similar clinical presentation. PATIENTS AND METHODS: Among 2086 patients with an acute coronary syndrome (ACS) undergoing coronary angiography over a 6.5-year period, 33 patients (1.6 %) with apical ballooning (AB) were identified (29 f, 4 m, median age 77 [68 - 80] years). AB patients were compared to 28 consecutive age and sex matched AMI patients (23 f, 5 m; 68 [56 - 76] years) undergoing successful PCI of the LAD with similar findings on LV angiography. We compared the ECG on admission, at the time of maximal T-wave inversion, before discharge and the longest QTc-interval. RESULTS: The number of leads showing ST-segment elevation (4 [3 - 6] vs. 5 [5 - 7]; p = 0,005) and the sum of the level of ST-segment elevation (0,7 [0,5 - 0,9] mV vs. 0,9 [0,7 - 1,5] mV; p = 0,002) was significantly greater in AMI. An abnormal Q wave at presentation was more frequent in AMI (21 % vs 79 %, p < 0.001) and persisted in half of AMI patients whereas the Q wave disappeared in all AB patients during follow-up (0 % vs. 61 %, p < 0.001). The QTc interval was significantly longer in AB (568 [521 - 614] ms vs. 471 [438 - 513] ms; p < 0,001). During follow-up, AB patients displayed significantly more leads with T wave inversion (8 [8 - 9] vs. 6 [5 - 8]; p < 0,001), and the sum of the level of T-wave inversion (2,9 [2,2 - 4,6] mV vs. 1,4 [0,9 - 2,3] mV; p < 0,001) was significantly larger than in AMI. A formula considering ST-segment elevation on admission and T-wave inversion as well as Q waves during follow-up allowed discrimination between AB and AMI (sensitivity 93 %, specificity 86 %). CONCLUSION: ECG findings in AB patients are significantly different from those in AMI patients. At initial presentation, the extent of ST-segment elevation and the number of abnormal Q waves are greater in AMI. During follow-up, no Q wave, a longer QTc interval and a greater extent of T-wave inversion are typical findings in AB patients. PMID- 18398791 TI - [Dermatoma complicating Herpes zoster during immunosuppression]. PMID- 18398792 TI - [Splenic rupture after colonoscopy]. AB - HISTORY AND CLINICAL FINDINGS: A 57-year-old woman was admitted with nonspecific pain in the left upper quadrant of the abdomen, radiating to her left shoulder, starting eight hours after a prophylactic colonoscopy. Her past surgical history consisted of a pylorus-preserving partial duodenopancreatectomy for chronic pancreatitis. INVESTIGATIONS: After a colon perforation had been excluded ultrasound and computed tomography revealed a covered splenic rupture. TREATMENT AND COURSE: Because the patient was hemodynamically stable she was observed in the intensive care unit with hemodynamic monitoring. When the patient became hemodynamically unstable an ultrasound revealed an increase of the perisplenic fluid. She immediately had an exploratory laparotomy and the spleen wrapped in a Vicryl net. Postoperatively the patient had an uncomplicated hospital stay and was discharged on postoperative day 7. CONCLUSION: Splenic rupture is a known but rare complication of colonoscopy. Early recognition and interdisciplinary management will assure successful management of this potentially life-threatening injury. PMID- 18398793 TI - [Complications after lung transplantation: gastroenterological, hematological and cardiovascular involvement]. AB - Intestinal perforation after lung transplantation is common in elderly patients with diverticulitis. The distal intestinal obstruction syndrome (DIOS) is a current complication in patients with cystic fibrosis. Myelosuppressive drugs as in most immunosuppressive regimens can cause cytopenia. Thrombotic microangiopathy is often caused by immunosuppressive regimen with a calcineurin inhibitor in combination with a proliferation signal-inhibitor. Although renal failure and neurological symptoms are often additional side effects of the immunosuppressive therapy there are no reliable data on CNI-free protocols. Studies have shown that administration of statins is associated with improved function and survival of lung allografts. PMID- 18398794 TI - [Botulism: prevention, clinical diagnostics, therapy and possible threat]. AB - In times of increased terrorist threat health professionals need to be prepared for bioterrorist events. The goal must be to give doctors an overview over the current state of knowledge and risk assessment of botulinum toxin. This review is based on Information gathered by a systematic analysis of the literature and by contacting experts. The toxicity of botulinum toxin exceeds any other known natural toxin. Clinical features of botulism consist of an acute, afebrile symmetrical and descending paralysis, regardless of the route of exposure, with normal mental status, sensory functions and electrolyte values. The initial diagnosis is often wrong in individual cases of botulism, but clusters with typical symptoms and two or more cases usually provide the diagnosis. Current treatment is primarily supportive care, respiratory support and antitoxin administration. Early application of antitoxin can limit the extent of the paralysis, but will not reverse it. Antitoxin for adult patients is of equine origin, while children in the USA can be treated with a recently developed human antitoxin. A pentavalent toxoid vaccine is available for persons at high risk of exposure. Botulinum toxin is easily extracted and ubiquitously available. These two features, together with the high toxicity, makes misuse easy. Misuse will continue to occur. Although a rare disease in Western Europe, botulism should be included in the differential diagnosis in patients with specific symptoms of paralysis. There is the potential threat of deliberate release of botulinum toxin. For this reason every outbreak of botulism must be assessed for any possible links to terrorism. PMID- 18398795 TI - [Nutrition in liver cirrhosis: diagnostic aspects and treatment]. AB - Malnutrition is very common in patients with liver cirrhosis and includes especially protein-calorie malnutrition. The pathophysiological reasons vary and are caused by metabolic modifications and characteristics of enteral absorption and digestion cause by the cirrhosis. Malnutrition contributes to overall mortality and complication rate of the chronic liver disease. An adequate daily energy and protein supply should be ensured in patients with liver cirrhosis, which is higher than that in the normal population, because of higher amino-acid turnover. Ascites may benefit from daily low-salt fluid intake. Nutritional substitution of vitamins and trace elements is indicated when symptoms of deficiency occur. PMID- 18398797 TI - [Patients with mild cognition impairment should not be treated with antidementia agents, except in studies]. PMID- 18398798 TI - [Sobering but hopeful!]. PMID- 18398799 TI - [Prevention of post-discharge suicide: an inventory]. AB - BACKGROUND: Research has shown that the first months after discharge from psychiatric inpatient care is a high risk period for suicide. However, there has been very little research into the prevention policies that mental health care centres pursue in order to reduce this risk. AIM: To draw up an inventory of the preventive activities that mental health services undertake to reduce the risk of suicide by psychiatric patients discharged from inpatient treatment. METHOD: Fifteen mental health care providers of 10 different departments of 4 mental health care organisations in the province North-Holland were asked in a personal interview about the policy of their institution regarding the prevention of suicide after discharge. results One out of 10 locations had a standard policy for the prevention of suicide after discharge from psychiatric care. Four locations had an informal policy and 5 an ad hoc policy. Differences were found in the views of mental health care providers regarding suicide prevention and the responsibility of mental health care centres for the prevention of post-discharge suicide. CONCLUSION: Only half of the mental health institutions employed a preventive policy with regard to post-discharge suicide. So far, the possibilities for prevention have not been fully utilised. PMID- 18398800 TI - [Beta-blockers and electroconvulsive therapy: a review]. AB - BACKGROUND: When patients with cardiovascular disorders undergo electroconvulsive therapy (ect) they sometimes have to be treated for tachycardia and high blood pressure. AIM: To describe the effects of beta-blockers on seizure duration and cardiovascular variables in patients undergoing ect. METHOD: Search for studies in Medline, with the keywords 'beta-adrenergic blocking agents' and 'electroconvulsive therapy'. Only articles based on randomised placebo-controlled investigations were included. results The search strategy produced 21 articles. These were assessed by all authors. Esmolol was the drug administered in most of the trials. Since seizure duration can influence the therapeutic effect of ect it is advisable to use bilateral electrode placement in patients with cardiovascular risk factors and to administer esmolol prior to seizure induction. CONCLUSION: The beta-blocker of choice for use during ect seems to be esmolol; it can shorten seizure duration, although the effect is probably dose-dependent. Esmolol is also the drug of choice in ect sessions for patients without cardiovascular risk factors but who develop prolonged hypertension or tachycardia. A possible alternative is labetalol, but its longer half-life is a disadvantage, particularly if it is administered in a high dose. So far, experience with landiolol is limited, but its short half-life, greater cardioselectivity and higher potency mean that it could be a promising alternative. PMID- 18398801 TI - [Augmentation with atypical antipsychotics for the treatment of patients with a therapy-resistant depression: a review]. AB - BACKGROUND: A considerable number of depressed patients are showing resistance to current drug treatment strategies. In such cases it is becoming increasingly common in clinical practice to augment an antidepressant with an atypical antipsychotic (aap). purpose To provide an overview of the scientific evidence for this new strategy, to explain the possible mechanisms of action and to assess the place that augmentation with an aap occupies in the treatment of therapy resistant non-psychotic depression. METHOD: Various search terms were used to locate relevant articles in Pubmed; these articles were examined for relevant references. results Only 6 randomised controlled trials were found, therefore 7 case reports and 10 open-label studies were included. There seems to be some evidence that augmentation with an aap, particularly olanzapine, is effective. One of the main advantages of this strategy is the fast response, namely within a few weeks or even within a week. In view of the lack of scientific support for augmentation with aaps, this strategy is advisable when other augmentation strategies have proved unworkable or are contraindicated. It must be exercised with caution because the combination therapy can have negative effects on the patient's glucose and lipid metabolism. CONCLUSION: Augmentation with aaps in treatment resistant depression may be a potential useful treatment strategy but its scientific evidence is insufficient to warrant inclusion within current guidelines. PMID- 18398802 TI - [Should the treatment of depression in the Netherlands be changed after the STAR*D study?]. AB - The main results of the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) study are discussed in order to find out whether the results should lead to changes in the way in which depression is treated in the Netherlands. The outcomes of the STAR*D study are not sufficient justification for altering the current recommendations about the successive steps to be taken according to the Dutch multidisciplinary guideline for the treatment of depression. PMID- 18398803 TI - [Carbon monoxide poisoning; psychotic depression brought on by a new kitchen]. AB - BACKGROUND: An 81-year-old woman was diagnosed with chronic carbon monoxide poisoning. Her symptoms closely resembled those of someone with depression, although some of her symptoms were atypical. Because carbon monoxide is a colourless, odourless gas, the cause of her condition remained unclear for several weeks. If psychiatrists are better informed about the affective and neuropsychological symptoms that frequently accompany carbon monoxide poisoning, they should be able to recognise the symptoms sooner. This in turn should lead to a better outcome for the patient. PMID- 18398804 TI - [Reaction to 'Is it advisable to combine electroconvulsive therapy (ECT) with psychotropics?']. PMID- 18398805 TI - Diabetes mellitus and vascular disease: targeting cardiometabolic risk. PMID- 18398806 TI - The importance of treating cardiometabolic risk factors in patients with type 2 diabetes. AB - Cardiometabolic risk factors are the combined vascular and metabolic components of risk that may lead to a cardiovascular event. There are numerous such factors. Underlying the concept of cardiometabolic risk is an association with excess visceral fat, leading to the dysregulation of the adipokines, the signalling proteins derived from adipose tissue. Changes in the levels of the adipokines - tumour necrosis factor-alpha, cholesteryl ester transfer protein and adiponectin, for example - can lead to alterations in insulin sensitivity and high-density lipoprotein cholesterol metabolism. At present, specific cardiometabolic risk factors are commonly managed on an individual basis. We are now moving from the era of single risk factor intervention, however, to multiple risk factor intervention in people at high cardiovascular risk, with the additional possibility of using new drug classes to target the underlying cardiometabolic problems more effectively. PMID- 18398807 TI - Elevated serum ferritin concentrations in prediabetic subjects. AB - BACKGROUND: Few data are available on the association of variables of the insulin resistance syndrome and serum ferritin, an indicator of body iron stores. We examined the relationship between serum ferritin levels and impaired fasting glucose, a pre-diabetes stage associated with insulin resistance, in this study. SUBJECTS & METHODS: One hundred and eighty seven people, including 91 subjects with impaired fasting glucose (IFG) and 96 healthy people who were well matched for age and sex, were enrolled. Body mass index (BMI) and blood pressure of the participants were measured and serum cholesterol, triglyceride, white blood cells (WBC) count, C-reactive protein (CRP) and ferritin were evaluated. All the data were analysed by t-test, chi2 test and analysis of variance. RESULTS: The IFG group had higher serum ferritin concentrations (85.5+/-6.6 microg/L vs. 49.4+/ 3.7 microg/L, p=0.001). A positive correlation was found between fasting plasma glucose and serum ferritin (r=0.29, p=0.001). Using multiple regression analysis, we found an association between serum ferritin and blood pressure (0.15, p=0.01), FPG (0.29, p=0.001), triglyceride (0.08, p=0.01) and cholesterol (0.07, p=0.03). The odds ratio for the association of IFG in male subjects with a high serum ferritin level was 8.3 (95% CI: 1.2-11.9, p=0.01) and for females was 3.06 (95% CI: 0.58-15, p=0.1). CONCLUSION: Based on the data from our study, aelevation in serum ferritin can be seen in pre-diabetes stage, before the occurrence of an overt diabetes mellitus. PMID- 18398808 TI - Aminoterminal pro-brain natriuretic peptide (NT-proBNP) and sleep-disordered breathing in morbidly obese females: a cross-sectional study. AB - Sleep-disordered breathing (SDB) is often encountered in morbid obesity (MO) in conjunction with insulin resistance (IR). Aminoterminal pro-brain natriuretic peptide (NT-proBNP) is a promising marker for left ventricular dysfunction (LVD) in MO. We sought to explore the factors that may influence the relationships of SDB and IR with NT-proBNP in MO women. We performed a cross-sectional pilot study involving 110 asymptomatic MO (44.5+/-0.7 kg/m2) young women. SDB risk was assessed using a modified version of the Berlin Questionnaire (BQ). IR was assessed using the homeostasis model assessment (HOMA) index and adiponectin levels. LVD was assessed using NT-proBNP and echocardiograms. In this study, NT proBNP levels and LVD increased significantly along the BQ strata. Multiple regression analysis identified BQ and log-transformed HOMA as the independent variables predicting as much as 48.0% of the variability of logNT-proBNP. In conclusion, NT-pro-BNP levels are independently predicted by SDB and IR in asymptomatic MO women. Larger prospective studies are warranted. PMID- 18398809 TI - Beneficial effects of strategies for primary prevention of diabetes on cardiovascular risk factors: results of the Indian Diabetes Prevention Programme. AB - In subjects with impaired glucose tolerance (IGT) who participated in the Indian Diabetes Prevention Programme (IDPP), abnormalities related to body mass index, waist circumference (WC), blood pressure (BP), lipid profile and electrocardiography were analysed (at baseline and third-year follow-up) in control, lifestyle modification (LSM), metformin (MET) and LSM + MET groups. At baseline, elevated levels of low-density lipoprotein cholesterol (LDL-C) showed the highest (78.6%) and total cholesterol (TC) showed the lowest (42%) prevalence. At follow-up, prevalence of hypertension (BP > or = 130/> or = 85 mmHg) had increased significantly in all groups. Cardiovascular abnormalities were lower in intervention groups, with the lowest rates in the MET group (p=0.013 vs. control); the LDL-C level decreased in intervention groups. In this programme, Asian Indian IGT subjects were observed to have a high prevalence of cardiovascular risk factors. LSM and MET had beneficial effects on the atherogenic phenotype of lipids but had no influence on blood pressure. PMID- 18398810 TI - Detection and characterisation of microcirculatory abnormalities in the skin of diabetic patients with microvascular complications. AB - The aim of this study was to characterise microvascular blood flow in the skin and to compare it with biomarkers of endothelial dysfunction and tissue inflammation in patients with type 2 diabetes with (n=20) or without (n=20) microvascular complications and 20 control subjects. Microvascular function was measured by laser Doppler velocimetry in combination with iontophoresis of acetylcholine (ACh) and sodium nitroprusside (SNP). Blood was collected for measurement of biomarkers including plasminogen activator inhibitor-1 (PAI-1), soluble intercellular adhesion molecule (sICAM), soluble vascular cell adhesion molecule (sVCAM) and high-sensitivity C-reactive protein (hsCRP). Both ACh and SNP responses fall progressively with the development of diabetes and microvascular complications. For the total cohort, there was a significant overall correlation between ACh and SNP response (r=0.7, p<0.0001), and this relationship was particularly strong in those with microvascular complications. There was a trend towards higher hsCRP levels across the three groups, but no difference in other biomarkers. Abnormalities of microvascular blood flow are evident in diabetes and become more marked with the development of microvascular complications. This relationship was similar to that shown by the marker of inflammation (hsCRP), but stronger than that pertaining to biomarkers of endothelial function. As both ACh and SNP responses are attenuated, the disturbance is not characteristic of endothelial dysfunction alone. PMID- 18398812 TI - Characterisation of atherosclerotic lesions with scanning electron microscopy (SEM) of wet tissue. AB - Atherosclerotic cardiovascular disease (CVD) is the universal leading cause of mortality and a major cause of morbidity. Additionally, the global epidemic of diabetes is associated with considerable cardiovascular mortality risk due to accelerated premature atherosclerosis. Development of effective therapies for atherosclerosis is dependent upon improved tools to assess atherosclerotic lesion progression in animal models. We present a novel technique that utilises scanning electron microscopy (SEM) for imaging wet biological specimens, thus enabling rapid and high-resolution imaging of atherosclerotic lesions. This wet SEM technique was used in an apoE-deficient mice model for morphological characterisation of early and advanced atherosclerotic lesions. Further demonstration of lipid-rich atherosclerotic lesions was carried out with osmium tetroxide staining for cholesterol. Gold immunolabelling of specific epitopes was applied in identification of the cellular and molecular components within the atherosclerotic lesions, namely foam cells, smooth muscle cells and collagen. The wet SEM technique demonstrates an accurate and detailed structural evaluation of the pathological process of atherosclerosis. Understanding the mechanisms that precipitate the atherosclerotic process, utilising this novel technique, may assist in the development of innovative therapeutic interventions for CVD management and prevention in the general population and in those with diabetes. PMID- 18398813 TI - Drinking modulates monocyte migration in healthy subjects: a randomised intervention study of water, ethanol, red wine and beer with or without alcohol. AB - Moderate alcohol consumption is associated with reduced cardiovascular mortality compared to non-consumption of alcohol and heavy drinking. Experimental data suggest a direct effect of alcohol on atherosclerotic lesion development. We assessed the effect of consumption of moderate amounts of alcoholic and non alcoholic beverages on monocyte migration, a crucial step in atherogenesis. Forty nine healthy men and women (aged 22-56 years) were enrolled in this randomised controlled trial. After wash-out, participants were assigned to either ethanol (concentration 12.5%), beer (5.6%) or red wine (12.5%) equivalent to 30 grams of ethanol per day (g/d) for men and 20 g/d for women, or to the same amount of de alcoholised beer or red wine, or to water. Monocyte migration was evaluated ex vivo using a modified Boyden chamber. Intake of ethanol or de-alcoholised red wine significantly reduced monocyte chemoattractant protein-1 (MCP-1)-induced monocyte migration by 58% (p<0.05; n=6) and 36% (p<0.05; n=7) and FMLP (N-formyl methionyl-leucyl-phenylalanine)-induced migration by 41% (p<0.05) and 36% (p<0.05), respectively. MCP-1 receptor expression was not affected by these interventions, as shown by flow cytometry. Short-term intervention with moderate amounts of ethanol and de-alcoholised red wine inhibits monocyte migration ex vivo. This might represent one mechanism by which alcoholic beverages lower cardiovascular risk. PMID- 18398811 TI - Metabolic syndrome in the Philippine general population: prevalence and risk for atherosclerotic cardiovascular disease and diabetes mellitus. AB - The objectives of this study were to determine the prevalence of metabolic syndrome (MS) and its component risk factors among Filipinos using three sets of criteria and to evaluate the association between MS and atherosclerotic cardiovascular disease and diabetes mellitus. The study utilised a multi-staged cluster sampling design. The prevalence of MS was found to be 11.9% by National Cholesterol Education Program/Adult Treatment Panel (NCEP/ATP III) criteria, 14.5% by International Diabetes Federation (IDF) criteria and 18.6% by NCEP/ATP III criteria modified by the American Heart Association/National Heart, Lung and Blood Institute (NCEP/ATP III-AHA/NHLBI) criteria. Low levels of high-density lipoprotein cholesterol (HDL-C) occurred in 60.2% of men and 80.9% of women. Abdominal obesity was noted in 17.7% of men and 35.1% of women. Blood pressure (BP) > or = 130/85 mmHg was seen in 33.3%, hypertriglyceridaemia in 20.6% and fasting blood sugar > or = 100 mg/dL (5.55 mmol/L) in 7.1%. Age-adjusted odds ratios showed that MS, by all three definitions, predisposed an individual to diabetes mellitus (DM) and stroke while MS by the IDF definition predisposed an individual to myocardial infarction (MI). Individuals with MS did not have a significant predisposition to angina and peripheral artery disease (PAD). Thus, the metabolic syndrome is common in Filipinos, with low HDL-C as the most prevalent component. The metabolic syndrome predisposes to diabetes mellitus and stroke, with a tendency to MI using the IDF criteria. PMID- 18398814 TI - Patients with the metabolic syndrome and a disturbed cortisol balance display more microalbuminuria. AB - The objective of this study was to investigate whether patients with the metabolic syndrome (MetS) and an imbalance in cortisol metabolism express increased urinary albumin excretion compared to those patients with metabolic syndrome alone. Seventy-four patients with MetS were evaluated using a low-dose dexamethasone suppression test (LDDST) to identify disturbed cortisol balance (cortisol levels > 50 nmol/L after LDDST). The level of albumin in the urine was also evaluated. Disturbed cortisol balance was found in 8% of all evaluated patients with MetS. Microalbuminuria was present significantly more often (p<0.01) in those patients with MetS and an imbalance in cortisol metabolism compared with patients suffering MetS alone (urine albumin: 210 mg/L vs. 26 mg/L, respectively, p<0.01). A substantial percentage of patients with MetS had inappropriate cortisol homeostasis. Of importance, excretion of urinary albumin was increased in these patients. This observation may indicate that this subgroup within the MetS population has a higher cardiovascular risk and possible increased endothelial dysfunction, with a subsequent need for stricter control to prevent cardiovascular morbidity and mortality. PMID- 18398815 TI - Blocking the renin-angiotensin-aldosterone system to prevent diabetes mellitus. AB - Type 2 diabetes mellitus (DM) is increasing around the world, and the public health impact of DM, driven largely by cardiovascular disease complications, underpins the importance of continued efforts toward primary prevention of DM. Only a few interventions have been shown to prevent DM, with none of them yet proven to improve cardiovascular risk commensurately. Accumulating evidence suggest that drugs that block the renin-angiotensin-aldosterone system (RAAS), many of which have proven cardiovascular disease (CVD) benefit, also have favourable effects on parameters of glucose metabolism and incident diabetes. Here we review the evidence accumulated to date from animal studies, clinical mechanistic studies and clinical trials regarding the effect of RAAS inhibition and incident DM. PMID- 18398816 TI - Charge-dependent and charge-independent contributions to ion-protein interaction. AB - The thermodynamic framework and recent molecular descriptions of protein-salt interactions are critically reviewed. A reanalysis of earlier and recent data describing the role of salts on the thermal stability of collagen I over a wide range of concentration evidences the complex encroachment of charge-dependent and charge-independent interactions. Charge-independent interactions, operationally quantified by dilution parameters and association constants, are found to be the overriding factor for the large stabilization observed with nonbinding salts and for the large destabilization observed when strong salt binding occurs. Charge dependent interactions, namely screening and selective ion adsorption, are instead the prevailing components at low ionic strength and nonisoelectric pH. PMID- 18398817 TI - Quantification of acetylcholine in microdialysate of subcutaneous tissue by hydrophilic interaction chromatography/tandem mass spectrometry. AB - It has recently been suggested that acetylcholine plays an important role in the modulation of tissue inflammation. In order to further understand the newly discovered cholinergic anti-inflammatory pathway, tracking the concentration changes of acetylcholine in tissue is required. This paper describes the development of a method coupling hydrophilic interaction chromatography with electrospray ionization tandem mass spectrometry (HILIC/ESI-MS/MS) for the separation and quantification of acetylcholine in microdialysis samples of normal rats and of rats with local inflammation. The separation of acetylcholine from interferential endogenous compounds and inorganic cations was achieved with a zwitterionic stationary phase column using isocratic elution. Low-energy collision-induced dissociation tandem mass spectrometric (CID-MS/MS) analysis was carried out in the positive ion mode using multiple reaction monitoring (MRM) of the following mass transitions: m/z 146 --> 87 for acetylcholine and m/z 155 --> 87 for the internal standard acetylcholine-D9. The limit of detection for acetylcholine was found to be 0.075 fmol on-column with a signal-to-noise ratio of 3:1. The lower limit of quantification was 0.25 fmol on-column. The calibration curves obtained for acetylcholine in blank microdialysates were linear in the ranges of 0.025-50 nM and 0.025-0.5 nM, with correlation coefficients equal to or greater than 0.9994 and 0.9969, respectively. The recoveries of acetylcholine for high (2 nM) and low (0.5 nM) concentrations were in the ranges of 90-96% and 95-109%, respectively. The coefficients of variation for intra-day and inter-day reproducibility were equal to or less than 7.3% and 10.4%, respectively. The method has been successfully applied in the measurement of acetylcholine in microdialysates from normal and inflamed rat tissue. PMID- 18398818 TI - A monitoring programme for 1,3-dichloropropene and metabolites in groundwater in five EU countries. AB - BACKGROUND: 1,3 Dichloropropene (1,3-D) is a preplanting soil fumigant for the control of cyst and free-living nematodes and is currently undergoing a resubmission under Annex 1 listing of Directive 91/414/EEC. The characteristics of 1,3-D are such that the risk of it or its soil metabolites leaching through the soil profile cannot be excluded. As such, groundwater monitoring programmes were established in five EU countries representing a wide range of agricultural, climatic and hydrogeological situations, covering a range of groundwater vulnerability scenarios. All monitoring was conducted in areas where there has been historical use of 1,3-D. RESULTS: Over 5000 groundwater samples were analysed for the presence of 1,3-D and its metabolites over a 2 year period. Almost all analyses (for parent and metabolites) yielded concentrations of <0.1 microg L(-1). There were just two detections of >0.1 microg L(-1) (0.12 microg L( 1) and 0.4 microg L(-1)) for the 3-chloroacrylic acid metabolite in shallow groundwater samples of the alluvial gravels of the River Tietar in the Caceres region of Spain. CONCLUSION: Groundwater monitoring programmes have been conducted in the EU in five countries. These have demonstrated that there is negligible contamination of groundwater with 1,3-D or its metabolites across a range of agroclimatic regions where 1,3-D is known to have been used for a number of years. Local scientific knowledge of geological features, hydrology, agricultural practice and specific local issues was essential to the conduct of the study. PMID- 18398819 TI - Genomic origin, processing and developmental expression of testicular outer dense fiber 2 (ODF2) transcripts and a novel nucleolar localization of ODF2 protein. AB - Outer dense fibers are a major constituent of the sperm tail and outer dense fiber 2 (ODF2) protein is one of their major components. ODF2 shares partial homology with cenexin 1 and cenexin 2, regarded as centriolar proteins. We show that ODF2 and cenexin 2 transcripts are the product of differential splicing of a single gene, designated Cenexin/ODF2 and that cenexin 1 is an incomplete clone of ODF2. ODF2 terminates in exon 20b whereas in cenexin 2 this exon is spliced out and translation terminates in exon 24. We demonstrate a transcriptional switch during rat testicular development, from somatic-type to testis-type ODF2 and cenexin transcripts during the onset of meiosis. The switch is completed when spermiogenesis is established. ODF2 immunoreactive sites were visualized in the acroplaxome, along the sperm tail and the centrosome-derived sperm head-to-tail coupling apparatus. An unexpected finding was the presence of ODF2 antigenic sites, but not cenexin antigenic sites, in the dense fibrillar component of the nucleolus of Sertoli cells, spermatogonia and primary spermatocytes. The characterization of the genomic origin, processing and developmental expression of ODF2 transcript isoforms and their protein products can help reconcile differences in the literature on the role of ODF2 and cenexin in the centrosome. Furthermore, the finding of ODF2 in the dense fibrillar component of the nucleolus suggests that this protein, in addition to its presence in sperm outer dense fibers and centrosome, highlights and adds to the nucleolar function during spermatogenesis and early embryogenesis. PMID- 18398820 TI - Expression profiling of CD133+ and CD133- epithelial cells from human prostate. AB - BACKGROUND: Recent evidence suggests that prostate stem cells in benign and tumor tissue express the cell surface marker CD133, but these cells have not been well characterized. The aim of our study was to gene expression profile CD133 expressing cells. METHODS: We analyzed CD133-positive (CD133+) and -negative (CD133-) sub-populations of high-integrin expressing epithelial cells isolated from benign human prostate tissue and hormone-refractory prostate cancer (HRPC). RESULTS: CD133+ cells freshly isolated from benign prostate tissue exhibited an expression profile characteristic of a putative stem/progenitor cell population, with transcripts involved in biological processes ranging from development and ion homeostasis to cell communication. The profile of CD133- cells was consistent with that of a transit amplifying population, suggesting up-regulated proliferation and metabolism. Comparison of benign populations to those from HRPC showed some similarities between CD133+ profiles but also revealed significant differences that provide a tumor-specific pattern, which included evidence of increased metabolic activity and active proliferation. Subsequently, we demonstrated protein expression of a number of candidate genes in these cell populations and in benign tissue. In a novel observation we also found expression of some of these markers in prostate tumors, including the oligodendrocyte lineage transcription factor OLIG1. CONCLUSIONS: This study provides a unique genome-wide molecular signature of CD133+ and CD133- human prostate epithelial cells. This will provide a valuable resource for prostate stem cell biology research and the identification of novel therapeutic targets for the treatment of prostate cancer. PMID- 18398821 TI - Genome-wide analysis identifies 16q deletion associated with survival, molecular subtypes, mRNA expression, and germline haplotypes in breast cancer patients. AB - Breast carcinomas are characterized by DNA copy number alterations (CNAs) with biological and clinical significance. This explorative study integrated CNA, expression, and germline genotype data of 112 early-stage breast cancer patients. Recurrent CNAs differed substantially between tumor subtypes classified according to expression pattern. Deletion of 16q was overrepresented in Luminal A, and a predictor of good prognosis, both overall and for the nonluminal A subgroups. The deleted region most significantly associated with survival mapped to 16q22.2, harboring the genes TXNL4B and DXH38, whose expression was strongly correlated with the deletion. The area most frequently deleted resided on 16q23.1, 3.5 MB downstream of the area most significantly associated with survival, and included the tumor suppressor gene ADAMTS18 and the cell recognition gene CNTNAP4. Whole genome association analysis identified germline single nucleotide polymorphisms (SNPs) and their corresponding haplotypes, residing on several different chromosomes, to be associated with deletion of 16q. The genes where these SNPs reside encode proteins involved in the extracellular matrix (CHST3 and SPOCK2), in regulation of the cell cycle (JMY, PTPRN2, and Cwf19L2) and chromosome stability (KPNB1). PMID- 18398822 TI - Chromosome 11 genomic changes in parathyroid adenoma and hyperplasia: array CGH, FISH, and tissue microarrays. AB - We have used a combination of gene expression profiling, array comparative genomic hybridization (aCGH), fluorescent in situ hybridization (FISH) and tissue microarrays (TMAs) to investigate chromosome 11 genetic changes in subsets of benign parathyroid tumors. Integration of gene expression profiling and aCGH was done using differential gene locus mapping analysis. We have identified three distinct relatively common chromosome 11 genomic changes in various subsets of parathyroid tumors. The simplest and least common of these genomic changes involves translocation of the CCND1 gene with subsequent strong CCND1 expression. This genetic change is essentially limited to parathyroid adenomas (8%), although expression of CCND1 without translocation is common in uremic hyperparathyroidism. Not surprisingly, deletion of the MEN1 locus at 11q13 or loss of a large portion or an entire chromosome 11 was a common finding. This particular genomic change appears to have a prominent effect on the overall results of gene expression profiling and was present in slightly less than one half of adenomas. Genomic changes in primary nonfamilial hyperplasia were for the most part restricted to 11q13 deletion or loss of chromosome 11. The third genomic change we identified was 11q23 deletion. This genetic change was relatively independent of other chromosome 11 changes and present in slightly less than one-half of adenomas. 11q23 deletion along with relatively strong CCND1 expression was common in uremic hyperparathyroidism. PMID- 18398823 TI - Microhomologies and interspersed repeat elements at genomic breakpoints in chronic myeloid leukemia. AB - Reciprocal translocation t(9;22) is central to the pathogenesis of chronic myeloid leukemia. Some authors have suggested that Alu repeats facilitate this process, but supporting analyses have been sparse and often anecdotal. The purpose of this study was to analyze the local structure of t(9;22) translocations and assess the relevance of interspersed repeat elements at breakpoints. Collected data have been further compared with the current models of DNA recombination, in particular the single-strand annealing (SSA) and the nonhomologous end joining (NHEJ) processes. We developed a protocol for the rapid characterization of patient-specific genomic junctions and analyzed 27 patients diagnosed with chronic myeloid leukemia. Sequence analysis revealed microhomologies at the junctions of 21 patients of 27, while interspersed repeats were of relevance (P < 0.05) in at least 16 patients. These findings are more frequent than expected and give an indication that the main mechanisms involved in the t(9;22) translocation are the SSA and NHEJ pathways, both playing a role. Furthermore, our report is consistent with microhomologies facilitating the joining of DNA ends in the translocation process, and with both Alu and a variety of other repeat sequences pairing nonhomologous chromosomes during the SSA pathway. PMID- 18398824 TI - A novel basal ganglia pathway forms a loop linking a vocal learning circuit with its dopaminergic input. AB - Dopamine has been implicated in mediating contextual modulation of motor behaviors and learning in many species. In songbirds, dopamine may act on the basal ganglia nucleus Area X to influence the neural activity that contributes to vocal learning and contextual changes in song variability. Neurons in midbrain dopamine centers, the substantia nigra pars compacta (SNc) and ventral tegmental area (VTA), densely innervate Area X and show singing-related changes in firing rate. In addition, dopamine levels in Area X change during singing. It is unknown, however, how song-related information could reach dopaminergic neurons. Here we report an anatomical pathway that could provide song-related information to the SNc and VTA. By using injections of bidirectionally transported fluorescent tracers in adult male zebra finches, we show that Area X and other song control nuclei do not project directly to the SNc or VTA. Instead, we describe an indirect pathway from Area X to midbrain dopaminergic neurons via a connection in the ventral pallidum (VP). Specifically, Area X projects to the VP via axon collaterals of Area X output neurons that also project to the thalamus. Dual injections revealed that the area of VP receiving input from Area X projects to the SNc and VTA. Furthermore, VP terminals in the SNc and VTA overlap with cells that project back to Area X. A portion of the arcopallium also projects to the SNc and VTA and could carry auditory information. These data demonstrate an anatomical loop through which Area X activity could influence its dopaminergic input. PMID- 18398825 TI - Organization of the songbird basal ganglia, including area X. AB - Area X is a songbird basal ganglia nucleus that is required for vocal learning. Both Area X and its immediate surround, the medial striatum (MSt), contain cells displaying either striatal or pallidal characteristics. We used pathway-tracing techniques to compare directly the targets of Area X and MSt with those of the lateral striatum (LSt) and globus pallidus (GP). We found that the zebra finch LSt projects to the GP, substantia nigra pars reticulata (SNr) and pars compacta (SNc), but not the thalamus. The GP is reciprocally connected with the subthalamic nucleus (STN) and projects to the SNr and motor thalamus analog, the ventral intermediate area (VIA). In contrast to the LSt, Area X and surrounding MSt project to the ventral pallidum (VP) and dorsal thalamus via pallidal-like neurons. A dorsal strip of the MSt contains spiny neurons that project to the VP. The MSt, but not Area X, projects to the ventral tegmental area (VTA) and SNc, but neither MSt nor Area X projects to the SNr. Largely distinct populations of SNc and VTA dopaminergic neurons innervate Area X and surrounding the MSt. Finally, we provide evidence consistent with an indirect pathway from the cerebellum to the basal ganglia, including Area X. Area X projections thus differ from those of the GP and LSt, but are similar to those of the MSt. These data clarify the relationships among different portions of the oscine basal ganglia as well as among the basal ganglia of birds and mammals. PMID- 18398826 TI - Quantitation of methylated hemoglobin adducts in a signature peptide from rat blood by liquid chromatography/negative electrospray ionization tandem mass spectrometry. AB - Hemoglobin adducts are often used as biomarkers for exposure to reactive chemicals in toxicology studies. Therefore, fast, sensitive, accurate, and reproducible methods for quantifying these protein adducts are key to evaluate test material dosimetry. A methodology has been developed for the quantitation of methylated hemoglobin adducts isolated from rats exposed to the model alkylating agent: methyl methane sulfonate (MMS). After 4 days of MMS exposure by oral gavage, hemoglobin was isolated from rat blood and digested with trypsin. The tryptic digestion solution was used for the adducted hemoglobin signature peptide quantitation via liquid chromatography/negative tandem mass spectrometry (LC/ESI MS/MS). The limit of quantitation (LOQ) for the methylated hemoglobin beta chain N-terminal signature peptide (MeVHLTDAEK) was 1.95 ng/mL (5.9 pmol/mg globin). The calibration curves were linear over a concentration range of 1.95 to 625 ng/mL, with a correlation coefficient R2 >0.998, accuracy of 85.8 to 119.3%, and precision of 0.9 to 19.4%. PMID- 18398827 TI - Investigation of cholangiocarcinoma associated NKG2D polymorphisms in colorectal carcinoma. PMID- 18398828 TI - The risk of extra-colonic, extra-endometrial cancer in the Lynch syndrome. AB - Persons with the Lynch syndrome (LS) are at high risk for cancer, including cancers of the small bowel, stomach, upper urologic tract (renal pelvis and ureter), ovary, biliary tract and brain tumors, in addition to the more commonly observed colorectal and endometrial cancers. Cancer prevention strategies for these less common cancers require accurate, age-specific risk estimation. We pooled data from 4 LS research centers in a retrospective cohort study, to produce absolute incidence estimates for these cancer types, and to evaluate several potential risk modifiers. After elimination of 135 persons missing crucial information, cohort included 6,041 members of 261 families with LS associated MLH1 or MSH2 mutations. All were either mutation carriers by test, probable mutation carriers (endometrial/colorectal cancer-affected), or first degree relatives of these. Among mutation carriers and probable carriers, urologic tract cancer (N = 98) had an overall lifetime risk (to age 70) of 8.4% (95% CI: 6.6-10.8); risks were higher in males (p < 0.02) and members of MSH2 families (p < 0.0001). Ovarian cancer (N = 72) had an lifetime risk of 6.7% (95% CI: 5.3-9.1); risks were higher in women born after the median year of birth (p < 0.008) and in members of MSH2 families (p < 0.006). Brain tumors and cancers of the small bowel, stomach, breast and biliary tract were less common. Urologic tract cancer and ovarian cancer occur frequently enough in some LS subgroups to justify trials to evaluate promising prevention interventions. Other cancer types studied occur too infrequently to justify strenuous cancer control interventions. PMID- 18398829 TI - Metastatic cancer cells with macrophage properties: evidence from a new murine tumor model. AB - Metastasis is the process by which cancer cells disseminate from the primary neoplasm and invade surrounding tissue and distant organs, and is the primary cause of morbidity and mortality for cancer patients. Most conventional cancer therapies are ineffective in managing tumor metastasis. This has been due in large part to the absence of in vivo metastatic models that represent the full spectrum of metastatic disease. Here we identify 3 new spontaneously arising tumors in the inbred VM mouse strain, which has a relatively high incidence of CNS tumors. Two of the tumors (VM-M2 and VM-M3) reliably expressed all of the major biological processes of metastasis to include local invasion, intravasation, immune system survival, extravasation and secondary tumor formation involving liver, kidney, spleen, lung and brain. Metastasis was assessed through visual organ inspection, histology, immunohistochemistry and bioluminescence imaging. The metastatic VM tumor cells also expressed multiple properties of macrophages including morphological appearance, surface adhesion, phagocytosis, total lipid composition (glycosphingolipids and phospholipids) and gene expression (CD11b, Iba1, F4/80, CD68, CD45 and CXCR4). The third tumor (VM NM1) grew rapidly and expressed properties of neural stem/progenitor cells, but was neither invasive nor metastatic. Our data indicate that spontaneous brain tumors can arise from different cell types in VM mice and that metastatic cancer can represent a disease of macrophage-like cells similar to those described in several human metastatic cancers. The new VM tumor model will be useful for defining the biological processes of cancer metastasis and for evaluating potential therapies for tumor management. PMID- 18398830 TI - Gene expression changes during HPV-mediated carcinogenesis: a comparison between an in vitro cell model and cervical cancer. AB - We used oligonucleotide microarrays to investigate gene expression changes associated with multi-step human papillomavirus type 16 (HPV16)-mediated carcinogenesis in vitro. Gene expression profiles in 4 early passage HPV16 immortalized human keratinocyte (HKc) lines derived from different donors were compared with their corresponding 4 late-passage, differentiation-resistant cell lines, and to 4 pools of normal HKc, each composed of 3 individual HKc strains, on Agilent 22 k human oligonucleotide microarrays. The resulting data were analyzed using a modified T-test coded in R to obtain lists of differentially expressed genes. Gene expression changes identified in this model system were then compared with gene expression changes described in published studies of cervical intraepithelial neoplasia (CIN) and cervical cancer. Common genes in these lists were further studied by cluster analysis. Genes whose expression changed in the same direction as in CIN or cervical cancer (concordant) at late stages of HPV16-mediated transformation in vitro formed one major cluster, while those that changed in the opposite direction (discordant) formed a second major cluster. Further annotation found that many discordant expression changes involved gene products with an extracellular localization. Two novel genes were selected for further study: overexpression of SIX1 and GDF15, observed during in vitro progression in our model system, was confirmed in tissue arrays of cervical cancer. These microarray-based studies show that our in vitro model system reflects many cellular and molecular alterations characteristic of cervical cancer, and identified SIX1 and GDF15 as 2 novel potential biomarkers of cervical cancer progression. PMID- 18398831 TI - Opposite impact of NKG2D genotype by lifestyle exposure to risk of aerodigestive tract cancer among Japanese. AB - It was reported that there are 2 haplotypes in natural killer complex (NKC) region. One of them could be divided by NKG2D polymorphism into 2 haplotype alleles (high and low natural killer (NK) cell activity) and were associated with overall cancer risks. However, its impact on a specific cancer is unclear. Therefore, by a case-control study, we analyzed the association between NKG2D genotype and aerodigestive tract cancer risk. Subjects were 502 aerodigestive tract cancer patients (276 with head and neck, 226 with esophageal) and 1,004 sex age matched noncancer controls. Exposures to 2 lifestyle factors, smoking and drinking, were evaluated by a self-administered questionnaire. The genotype of NKG2D was determined by the TaqMan method, and its impact was assessed by multivariable logistic regression models. Association strength was measured by the odds ratio (OR) and its confidence intervals (CI). An overall analysis revealed no statistically significant association between NKG2D genotype and the risk of aerodigestive tract cancer. However, we found protective effects of G allele among never smokers (OR 0.35; 95% CI 0.15-0.84) and never drinkers (0.42; 0.19-0.94). In contrary, increased risks were observed for G allele among heavy smokers (5.92; 3.23-10.85) and heavy drinkers (4.13; 2.29-7.47). Interactions between NKG2D genotype and lifestyle exposure were statistically significant (interaction p = 0.001 for smoking, 0.005 for drinking). The same trends were observed in both sexes, and in head and neck cancer and esophageal cancer independently. These results suggest an opposite impact of NKG2D genotype by lifestyle exposure to the risk of aerodigestive tract cancer among a Japanese population. PMID- 18398833 TI - Age-related changes in the human thymus studied with scanning electron microscopy. AB - This investigation describes some morphological, age-related changes in different compartments and cells of the human thymus. Scanning electron microscopic observations were done on human thymus samples obtained from four young and eight elderly patients during thoracic surgery and/or diagnostic biopsy of the thymus, after receiving authorization from the Ethical Committee of our university. The morphological data were submitted to quantitative image analysis so as to obtain quantitative results. Subsequently, the related values were used for statistical analysis. Our findings demonstrate that (1) all thymus compartments (subcapsular spaces, cortical, medullar, thymus microenvironment) contain numerous thymocytes even after the thymus has aged. (2) In older humans, residual thymus lymphoid islets contain, in addition to fatty cells and/or fibrous cells, also the same types of resident and nonresident cells (permanent and moving cells) that are found in young and adult subjects. (3) Endothelial cells of thymus microvessels contain numerous gaps. These gaps are tight in young subjects and become loose with age. (4) Thymocytes, in older subjects, are always found near these loose endothelial gaps of thymus microvessels. (5) While thymus cortical microvessels are provided with pericytes and/or periarteriolar spaces, microvessels of the thymus medullar are free of such spaces. Our morphological and quantitative results lead us to consider the possibility that the thymus fraction of resident and permanent cells (including thymocytes and reticular epithelial cells) is larger in younger subjects compared with older ones. The endothelial loose gaps of thymus microvessels, in older subjects, can allow the bidirectional transit of thymocytes through the wall of the said microvessels. PMID- 18398832 TI - Evolution of decitabine development: accomplishments, ongoing investigations, and future strategies. AB - Decitabine (5-aza-2'-deoxycytidine) is a hypomethylating agent with a dual mechanism of action: reactivation of silenced genes and differentiation at low doses, and cytotoxicity at high doses. The original studies in the 1980s used decitabine as a classical anticancer drug, at its maximum clinically tolerated dose, 1500 to 2500 mg/m(2) per course. At these doses, decitabine was found to be active in leukemia, but was associated with delayed and prolonged myelosuppression. After a better understanding of epigenetics in cancer and the role of decitabine in epigenetic (hypomethylating) therapy was gained, it was reevaluated at approximately 1/20th of the previous doses (ie, at 'optimal biologic' doses that modulate hypomethylation). In these dose schedules of decitabine (100 to 150 mg/m(2) per course), the drug was found to be active with manageable side effects in patients with myelodysplastic syndromes (MDS) and other myeloid tumors. Optimizing dosing schedules of decitabine to maximize hypomethylation (low dose, high dose intensity, and multiple cycles) have further improved results, suggesting that decitabine is an active therapy that alters the natural course of MDS. Combination therapies that augment the epigenetic effect of decitabine will likely improve responses and extend its use for the treatment of other malignancies. PMID- 18398834 TI - Hibernation as a far-reaching program for the modulation of RNA transcription. AB - In eukaryotic cells, pre-mRNAs undergo several transformation steps to generate mature mRNAs ready to be exported to the cytoplasm. The molecular and structural apparatus for mRNA production is generally able to promptly respond to variations of metabolic demands. Hibernating mammals, which periodically enter a hypometabolic state, represent an interesting physiological model to investigate the adaptive morpho-functional modifications of the pre-mRNA transcriptional and processing machinery under extreme metabolic conditions. In this study, the subnuclear distribution of some transcriptional, splicing, and cleavage factors was investigated by ultrastructural immunocytochemistry in cell nuclei of the liver (a highly metabolizing organ involved in multiple regulatory functions) and the brown adipose tissue (responsible for nonshivering thermogenesis) from euthermic, hibernating, and arousing hazel dormice (Muscardinus avellanarius). Our observations demonstrate that, during hibernation, transcriptional activity significantly decreases and pre-mRNA processing factors undergo an intranuclear redistribution moving to domains usually devoid of such molecules; moreover, in hepatocytes, there is a preferential accumulation of pre-mRNAs at the splicing stage, whereas, in brown adipocytes, pre-mRNAs are mainly stored at the cleavage stage. Upon arousal, the pre-mRNAs at the cleavage stage are immediately utilized, while the maturation of pre-mRNAs at the splicing stage seems to be restored before transcription had taken place. Our data suggest a programmed intranuclear reorganization of the RNA maturation machinery aimed at efficiently and rapidly restoring the pre-mRNA processing, and, consequently, the specific cellular activities upon arousal. Once again natural hibernation appears as a highly programmed hypometabolic state rather than a simple fall of metabolic and physiological functions. PMID- 18398835 TI - High resolution X-ray phase contrast synchrotron imaging of normal and ligation damaged rat sciatic nerves. AB - This study was performed to apply synchrotron radiation (SR) imaging to a neuropathologic evaluation technique after treatment of peripheral nerve blocks. A phase contrast synchrotron images of normal and ligation damaged rat sciatic nerve were obtained with an 8 KeV monochromatic beam and 20-mum thick CsI(TI) scintillation crystal. The visual image was magnified using a 20x microscope objective and captured using an analog CCD camera. Obtained images were compared with conventional light microscopic findings from the same nerve samples. By using an edge enhancement effect of phase const with SR, we could easily discriminate each nerve fiber and identify the arrangement of nerve fibers within a whole thickness (about 1 mm in diameter) of peripheral nerve without sectioning and fixation. The composite SR image of a ligation damaged rat sciatic nerve sample showed that the response to nerve injury was different on each side of the site of injury. The SR image of damaged distal lesion showed destruction of neural microarchitecture and typical extensive Wallerian degeneration of nerve fibers as clearly as histologic image. We could get very detailed morphologic data for Wallerian degeneration of nerve fibers by using the SR imaging technique. We believe that the phase contrast synchrotron imaging has great potential as an imaging tool in the bioscience and medical science. PMID- 18398837 TI - Identification of novel DNA methylation markers in cervical cancer. AB - Testing for DNA methylation has potential in cancer screening. Most previous studies of DNA methylation in cervical cancer used a candidate gene approach. The aim our study was to identify novel genes that are methylated in cervical cancers and to test their potential in clinical applications. We did a differential methylation hybridization using a CpG island (CGI) microarray containing 8640 CGI tags to uncover methylated genes in squamous cell carcinomas (SCC) of the uterine cervix. Pooled DNA from cancer tissues and normal cervical swabs were used for comparison. Methylation-specific polymerase chain reaction, bisulfite sequencing and reverse transcription polymerase chain reaction were used to confirm the methylation status in cell lines, normal cervices (n = 45), low-grade lesions (n = 45), high-grade lesions (HSIL; n = 58) and invasive squamous cell carcinomas (SCC; n = 22 from swabs and n = 109 from tissues). Human papillomavirus (HPV) was detected using reverse line blots. We reported 6 genes (SOX1, PAX1, LMX1A, NKX6 1, WT1 and ONECUT1) more frequently methylated in SCC tissues (81.5, 94.4, 89.9, 80.4, 77.8 and 20.4%, respectively) than in their normal controls (2.2, 0, 6.7, 11.9, 11.1 and 0%, respectively; p < 0.0001). Parallel testing of HPV and PAX1 methylation in cervical swabs confers an improved sensitivity than HPV testing alone (80% vs. 66%) without compromising specificity (63% vs. 64%) for HSIL/SCC. Testing PAX1 methylation marker alone, the specificity for HSIL/SCC is 99%. The analysis of these novel DNA methylations may be a promising approach for the screening of cervical cancers. PMID- 18398838 TI - Expression of CD175 (Tn), CD175s (sialosyl-Tn) and CD176 (Thomsen-Friedenreich antigen) on malignant human hematopoietic cells. AB - The expression of the histo-blood group carbohydrate structures T-nouvelle (Tn, CD175), sialylated Tn (CD175s) and the Thomsen-Friedenreich disaccharide (TF, CD176) on human leukemia cell lines was analyzed by their reactivity with specific monoclonal antibodies in flow cytometry, immunohistology and immunoprecipitation. Expression of sialylated CD176 was evaluated by comparative immunostaining with anti-CD176 antibodies before and after sialidase treatment. While only few cell lines expressed unmasked CD176, sialylated CD176 was present on all hematopoietic cell lines and native lymphocytes examined. CD175 and CD175s are preferentially expressed on erythroblastic leukemia cell lines. CD175s expression in these cells is consistent with the transcription of the gene encoding the key enzyme alpha2,6-sialyltransferase (hST6GalNAc1). The staining intensity was reduced after methanol pretreatment of cells, indicating that these glycans are partially expressed as constituents of glycosphingolipids. Immunoprecipitation and subsequent Western blotting revealed a series of distinct high molecular glycoproteins as carriers for these carbohydrate antigens. CD34 was identified as major carrier of CD176 by immunoprecipitation and microsequencing on a KG-1 subline enriched for CD176 expression. Incubation of several CD176-positive cell lines with anti-CD176 antibodies induced apoptosis of these cells, an effect not observed with anti-CD175/CD175s antibodies. Since the presence of naturally occurring anti-CD176 antibodies may represent a mechanism of immunosurveillance against CD176-positive tumor cells, we propose that sialylation of surface-expressed CD176--among other functions--protects against apoptosis. PMID- 18398839 TI - Persistent downregulation of the pancarcinoma-associated epithelial cell adhesion molecule via active intranuclear methylation. AB - The epithelial cell adhesion molecule (EpCAM) is expressed at high levels on the surface of most carcinoma cells. SiRNA silencing of EpCAM expression leads to reduced metastatic potential of tumor cells demonstrating its importance in oncogenesis and tumor progression. However, siRNA therapy requires either sequential delivery or integration into the host cell genome. Hence we set out to explore a more definite form to influence EpCAM gene expression. The mechanisms underlying the transcriptional activation of the EpCAM gene, both in normal epithelial tissue as well as in carcinogenesis, are poorly understood. We show that DNA methylation plays a crucial role in EpCAM expression, and moreover, active silencing of endogenous EpCAM via methylation of the EpCAM promoter results in a persistent downregulation of EpCAM expression. In a panel of carcinoma derived cell lines, bisulfite analyses showed a correlation between the methylation status of the EpCAM promoter and EpCAM expression. Treatment of EpCAM negative cell lines with a demethylating agent induced EpCAM expression, both on mRNA and protein level, and caused upregulation of EpCAM expression in an EpCAM positive cell line. After delivery of the DNA methyltransferase M.SssI into EpCAM positive ovarian carcinoma cells, methylation of the EpCAM promoter resulted in silencing of EpCAM expression. SiRNA-mediated silencing remained for 4 days, after which EpCAM re-expression increased in time, while M.SssI-mediated downregulation of EpCAM maintained through successive cell divisions as the repression persisted for at least 17 days. This is the first study showing that active DNA methylation leads to sustained silencing of endogenous EpCAM expression. PMID- 18398841 TI - A celecoxib derivative inhibits focal adhesion signaling and induces caspase-8 dependent apoptosis in human acute myeloid leukemia cells. AB - Most acute myeloid leukemias (AMLs), including those with c-Kit or FLT3 mutations, show enhanced anchorage independent growth associated with constitutive activation of focal adhesion proteins. Moreover, these alterations increase cell survival, inhibit apoptosis and are associated with poor prognosis and resistance to chemotherapy. Therefore, the induction of apoptosis by selective inhibition of focal adhesion signaling may represent a novel anti-AML therapy. Here, we have evaluated the antitumor effect and the mechanism of action of celecoxib and E7123, a non-Cox-2 inhibitor derivative, in a panel of human AML cell lines and bone marrow mononuclear cells from AML patients. Both compounds induce cell death by inhibiting focal adhesion signaling through p130Cas, FAK and c-Src, leading to caspase-8 dependent apoptosis. This mechanism of action differs from that of classical cytotoxic drugs or of other targeted therapies, and is amenable to rational drug development. Therefore, both drugs could be developed as AML therapeutics; nevertheless, E7123 shows more activity than celecoxib against AML cells, and may not present its Cox-2 dependent cardiovascular toxicity. Finally, our results support the evaluation of celecoxib in AML patients, and the preclinical evaluation of E7123, before its possible clinical testing. PMID- 18398842 TI - Heregulin-induced activation of ErbB3 by EGFR tyrosine kinase activity promotes tumor growth and metastasis in melanoma cells. AB - ErbB3 receptor tyrosine kinase has been shown to induce tumor progression in several types of cancer through heterodimerization with ErbB2. However, the role of ErbB3 and its ligand heregulin (HRG) in tumor metastasis remains poorly understood. In the present study, we tried to clarify their contributions to the metastasis of ErbB3-overexpressing B16-BL6 melanoma cells. Stimulation with HRG induced phosphorylation of ErbB3 and metastatic properties including MMP-9 expression, invasion, adhesion and experimental lung metastasis in vivo. These cellular responses were blocked by inhibiting the tyrosine kinase activity of EGFR with PD153035. In addition, phosphorylation of EGFR was rapidly induced by HRG, suggesting that EGFR is a possible heterodimeric counterpart of ErbB3. RNA interference demonstrated that subcutaneous tumor growth and angiogenesis was attenuated by inactivation of ErbB3 in cancer cells. Although experimental pulmonary metastasis was not affected by the knockdown of ErbB3, spontaneous metastasis was, even when primary tumors in the foot pad were amputated at a similar size. These results indicate that HRG-induced activation of ErbB3 via EGFR promotes tumor growth and metastasis of melanoma cells. PMID- 18398844 TI - How basal are triple-negative breast cancers? AB - The basal molecular subtype of breast cancer (BC) is defined by the mRNA expression pattern of an intrinsic approximately 500-gene set. It is the most homogeneous subtype in transcriptional terms, and one of the most aggressive in prognostic terms. Clinical trials testing new systemic therapeutic strategies have been launched in basal BCs. Although no proof of evidence has yet been reported, basal tumors are currently assimilated to and selected as triple negative (TN) BCs in these trials because of their frequent immunohistochemical (IHC) negativity for hormone and ERBB2 receptors. Here, we have assessed the degrees of correlation and of homogeneity of the TN phenotype (IHC-based definition) and the basal subtype (gene expression-based definition). We analyzed 172 TN BCs defined by gene expression profile as basal (123 cases) and nonbasal (49 cases). Conversely, 160 tumors were defined as basal by their gene expression profile and included 123 TN and 37 non-TN samples. Uni- and multivariate analyses revealed that TN BCs represent a more heterogeneous group than basal BCs, including basal and nonbasal tumors very different both at the histoclinical and molecular level, notably for mRNA expression of molecules targeted by specific therapies under evaluation in clinical trials. These results call for caution in the interpretation of ongoing trials and selection of patients in future trials. They also warrant the identification of molecular markers for basal BCs more clinically applicable than gene expression profiles. PMID- 18398845 TI - Investigating the specificity of periosteal reactions in pathology museum specimens. AB - The relationship between periosteal new bone formation and a number of infectious and metabolic conditions frequently seen in archeological human skeletal remains was investigated by studying human long bones demonstrating periosteal new bone formation archived in two London, UK, pathology museums: the St. George's Hospital Pathology Museum and the Hunterian Museum. The samples were subjected to macroscopic and radiographic analysis to determine if the characteristics of their periosteal lesions were specific to the corresponding disease states. The results demonstrated that no qualitative or quantitative characteristics of the periosteal reactions emerged that were specific to individual disease states. It was established that disease progression, rather than disease type, was the most important determinant of periosteal lesion appearance. A critical analysis of the bioarcheology literature pertaining to the recording and interpretation of periosteal reactions determined that the varied pathogenesis of periosteal new bone formation has been largely ignored in favor of a diagnosis of "nonspecific infection." Assumptions regarding the infectious etiology of periosteal lesions have become embedded into the bioarcheology literature potentially skewing the results of skeletal population-based paleoepidemiological studies. PMID- 18398843 TI - Nox1 is over-expressed in human colon cancers and correlates with activating mutations in K-Ras. AB - The NADPH-oxidase 1 (Nox1) is a homolog of gp91phox, the catalytic subunit of the phagocyte superoxide-generating NADPH-oxidase. Nox1 is expressed in normal colon epithelial cells and in colon tumor cell lines, and overexpression in model cells has been implicated in stimulation of mitogenesis and angiogenesis and inhibition of apoptosis. This suggests that aberrant expression of Nox1 could contribute to the development of colorectal cancer. Herein, we examine the expression of Nox1 mRNA in 24 colon tumors of various stages compared with paired adjacent normal tissue from the same patient, and correlate expression with some common mutations associated with colon cancer. Nox1 was overexpressed compared with paired normal tissue in 57% of tumors as early as the adenoma stage, with no correlation of expression level with tumor stage. Overexpression of Nox1 mRNA correlated with Nox1 protein levels assessed by immunofluorescence and immunohistochemistry with an antibody specific for Nox1. There was a strong correlation between Nox1 mRNA level and activating mutations in codons 12 and 13 of K-Ras. Eighty percent (8/10) of tumors with codons 12 and 13 mutations had a 2-fold or more increase in Nox1 mRNA, and 70% (7/10) had a 5-fold or greater increase. Transgenic mice expressing K-Ras(G12V) in the intestinal epithelium also expressed markedly elevated Nox1 in both small and large intestine. There was no correlation between inactivating mutations in the tumor suppressor p53 and Nox1 expression. We conclude that Nox1 mRNA and protein are overexpressed in colon cancer and are strongly correlated with activating mutations in K-Ras. PMID- 18398846 TI - A quantitative and descriptive approach to morphological variation of the endocranial base in modern humans. AB - The cranial base is one of the major foci of interest in functional craniology. The evolution and morphogenesis of this structure are still poorly known and rather controversial because of multifactorial influences and polyphasic stages. Endocranial dynamics are associated anteriorly with the upper facial structures, laterally with the mandibular system and midsagittally with brain development. In the present study, we investigated the endocranial morphology of modern humans using 3D landmark-based approaches, i.e. geometric morphometrics and Euclidean distance matrix analysis. The structure of endocranial variation is poorly integrated, with only weak reciprocal influences among the three fossae. Some major variations are associated with changes in the posterior fossa, with possible consequences on the anterior areas. These main patterns of integration are hypothesized to be influenced by the connective tensors of the dura layers. Static allometry and sex differences are largely related to the ontogenetic sequences, characterized by early maturation of the anterior fossa with respect to the middle and posterior regions (i.e., relatively shorter posterior part of the planum sphenoideum and vertical lengthening of the clivus in males). The relative independence between the endocranial fossae, as well as their structural connection through the meningeal tensors, must be carefully considered in studies on the evolutionary dynamics, since they lead to mosaic changes through phylogeny. PMID- 18398847 TI - Characterization and quantitative analysis of surfactants in textile wastewater by liquid chromatography/quadrupole-time-of-flight mass spectrometry. AB - A method based on the application of ultra-performance liquid chromatography (UPLC) coupled to hybrid quadrupole-time-of-flight mass spectrometry (QqTOF-MS) with an electrospray (ESI) interface has been developed for the screening and confirmation of several anionic and non-ionic surfactants: linear alkylbenzenesulfonates (LAS), alkylsulfate (AS), alkylethersulfate (AES), dihexyl sulfosuccinate (DHSS), alcohol ethoxylates (AEOs), coconut diethanolamide (CDEA), nonylphenol ethoxylates (NPEOs), and their degradation products (nonylphenol carboxylate (NPEC), octylphenol carboxylate (OPEC), 4-nonylphenol (NP), 4 octylphenol (OP) and NPEO sulfate (NPEO-SO4). The developed methodology permits reliable quantification combined with a high accuracy confirmation based on the accurate mass of the (de)protonated molecules in the TOFMS mode. For further confirmation of the identity of the detected compounds the QqTOF mode was used. Accurate masses of product ions obtained by performing collision-induced dissociation (CID) of the (de)protonated molecules of parent compounds were matched with the ions obtained for a standard solution. The method was applied for the quantitative analysis and high accuracy confirmation of surfactants in complex mixtures in effluents from the textile industry. Positive identification of the target compounds was based on accurate mass measurement of the base peak, at least one product ion and the LC retention time of the analyte compared with that of a standard. The most frequently surfactants found in these textile effluents were NPEO and NPEO-SO4 in concentrations ranging from 0.93 to 5.68 mg/L for NPEO and 0.06 to 4.30 mg/L for NPEO-SO4. AEOs were also identified. PMID- 18398848 TI - Combined neonatal stress and young-adult glucocorticoid stimulation in rats reduce BDNF expression in hippocampus: effects on learning and memory. AB - Epidemiological studies suggest that multiple developmental disruptions are involved in the etiology of psychiatric illnesses including schizophrenia. In addition, altered expression of brain-derived neurotrophic factor (BDNF) has been implicated in these illnesses. In the present study, we examined the combined long-term effect of an early stress, in the form of maternal deprivation, and a later stress, simulated by chronic young-adult treatment with the stress hormone, corticosterone, on BDNF expression in the hippocampus of rats. To assess whether there were behavioral effects, which may correlate with the BDNF changes, learning and memory was tested in the Y-maze test for short term spatial memory, the Morris water maze for long-term spatial memory, and the T-maze test for working memory. Four groups of rats received either no stress, maternal deprivation, corticosterone treatment, or both. Dorsal hippocampus sections obtained from parallel groups were used for BDNF mRNA in situ hybridization. Rats which had undergone both maternal deprivation and corticosterone treatment displayed a unique and significant 25-35% reduction of BDNF expression in the dentate gyrus (DG), and similar trends in the CA1 and CA3 regions of the hippocampus. These "two-hit" animals exhibited a learning delay in the Morris water maze test, a marked deficit in the Y-maze, but little change in the T-maze test. However, some aspects of cognition were also altered in rats with either maternal deprivation or corticosterone treatment. This study demonstrates a persistent effect of two developmental disruptions on BDNF expression in the hippocampus, with parallel, but not completely correlative changes in learning and memory. PMID- 18398850 TI - Two case studies illustrating how relatively selective hippocampal lesions in humans can have quite different effects on memory. AB - Two patients, with magnetic resonance imaging (MRI)-confirmed relatively selective hippocampal damage, showed distinct patterns of performance on tests of recall, item recognition, and associative recognition. Patient AC showed a mean bilateral volume reduction of the hippocampus of 28%, but displayed no memory deficit. Both recall and recognition memory were unimpaired. In contrast, patient PR, who showed a mean bilateral hippocampal volume reduction of 59%, was more consistently impaired on recall than recognition tests, although his recognition scores were highly variable. Patients AC and PR illustrate how variable the memory deficit following seemingly selective hippocampal damage can be in humans. They highlight the need for more sophisticated imaging in future studies if the human hippocampus' role in memory is to be fully identified. PMID- 18398849 TI - Altered morphology of hippocampal dentate granule cell presynaptic and postsynaptic terminals following conditional deletion of TrkB. AB - Dentate granule cells play a critical role in the function of the entorhinal hippocampal circuitry in health and disease. Dentate granule cells are situated to regulate the flow of information into the hippocampus, a structure required for normal learning and memory. Correspondingly, impaired granule cell function leads to memory deficits, and, interestingly, altered granule cell connectivity may contribute to the hyperexcitability of limbic epilepsy. It is important, therefore, to understand the molecular determinants of synaptic connectivity of these neurons. Brain-derived neurotrophic factor and its receptor TrkB are expressed at high levels in the dentate gyrus (DG) of the hippocampus, and are implicated in regulating neuronal development, neuronal plasticity, learning, and the development of epilepsy. Whether and how TrkB regulates granule cell structure, however, is incompletely understood. To begin to elucidate the role of TrkB in regulating granule cell morphology, here we examine conditional TrkB knockout mice crossed to mice expressing green fluorescent protein in subsets of dentate granule cells. In stratum lucidum, where granule cell mossy fiber axons project, the density of giant mossy fiber boutons was unchanged, suggesting similar output to CA3 pyramidal cell targets. However, filopodial extensions of giant boutons, which contact inhibitory interneurons, were increased in number in TrkB knockout mice relative to wildtype controls, predicting enhanced feedforward inhibition of CA3 pyramidal cells. In knockout animals, dentate granule cells possessed fewer primary dendrites and enlarged dendritic spines, indicative of disrupted excitatory synaptic input to the granule cells. Together, these findings demonstrate that TrkB is required for development and/or maintenance of normal synaptic connectivity of the granule cells, thereby implying an important role for TrkB in the function of the granule cells and hippocampal circuitry. PMID- 18398851 TI - Decreased hippocampal cell proliferation correlates with increased expression of BMP4 in the APPswe/PS1DeltaE9 mouse model of Alzheimer's disease. AB - Alterations in hippocampal cell proliferation have been identified in transgenic (tg) mouse models of Alzheimer's disease (AD); however, relatively little is known about the underlying mechanisms. Previously, we have demonstrated that endogenous level of BMP4 in the dentate gyrus (DG) affects hippocampal cell proliferation in a pentylentetrazol kindling-induced epilepsy model. In the present study, we evaluated hippocampal cell proliferation and BMP4 mRNA level in the APPswe/PS1DeltaE9 tg mouse, a well-established mouse model in which coexpression of familial AD-linked APP "Swedish" (APPswe) and PS1DeltaE9 polypeptide variants leads to Abeta deposition throughout the hippocampus and cortex. The number of bromodeoxyuridine (BrdU)-labeled cells in the DG subgranular zone (DG-SGZ) of 9- and 12-month-old APPswe/PS1DeltaE9 tg mice was markedly reduced compared with age-matched nontransgenic littermates, whereas, the BMP4 mRNA level was significantly increased in the tg mice. There was a significant correlation between the increased BMP4 mRNA expression and the decreased number of BrdU labeled cells. After effectively blocking the expression of endogenous BMP4 with antisense oligodeoxynucleotides (ASODN), the decrease in hippocampal cell proliferation in the DG-SGZ and hilus of 9- and 12-month-old tg mice was reversed. These findings suggest that the increased expression of BMP4 mRNA within the DG of the hippocampus may contribute to the decrease in cell proliferation in APPswe/PS1DeltaE9 tg mice. PMID- 18398852 TI - Transient 23-30 Hz oscillations in mouse hippocampus during exploration of novel environments. AB - The hippocampus is a key brain structure for the encoding of new experiences and environments. Hippocampal activity shows distinct oscillatory patterns, but the relationships between oscillations and memory are not well understood. Here we describe bursts of hippocampal approximately 23-30 Hz (beta2) oscillations in mice exploring novel, but not familiar, environments. In marked contrast to the relatively invariant approximately 8 Hz theta rhythm, beta2 power was weak during the very first lap of the novel environment, increased sharply as the mice reencountered their start point, then persisted for only a few minutes. Novelty evoked oscillations reflected precise synchronization of individual neurons, and participating pyramidal cells showed a selective enhancement of spatial specificity. Through focal viral manipulations, we found that novelty-evoked oscillations required functional NMDA receptors in CA3, a subregion critical for fast oscillations in vitro. These findings suggest that beta2 oscillations indicate a hippocampal dynamic state that facilitates the formation of unique contextual representations. PMID- 18398853 TI - Structural propensities in the heme binding region of apocytochrome b5. I. Free peptides. AB - The water-soluble domain of rat microsomal cytochrome b5 is a small globular hemoprotein. Under native conditions, the apoprotein consists of a well-folded hydrophobic core and a 42-residue loop, which is substantially disordered in solution. Association with the heme cofactor causes the loop to organize into a second well-folded hydrophobic core encompassing four short helices, H2-H5. Of these, H3 and H4 are recognized as intrinsically disordered by algorithms that analyze primary structures for folding propensities. Three peptides, spanning H2 H5, H2-H3, and H4-H5, were designed, synthesized, and characterized to identify local structural preferences in the isolated loop. In addition, two replacements (D60R and N57P, which are known to stabilize holocytochrome b5) were introduced individually in the H4-H5 peptide. Helical content measured by nuclear magnetic resonance and far-UV circular dichroism spectroscopy in solutions of 2,2,2 trifluoroethanol revealed that H4 possessed a lower propensity to form the holoprotein structure than H3. Both replacements in H4 resulted in measurable changes in observed overall helical propensities. It was concluded that the prediction of intrinsic disorder was reliable. Furthermore, the stability of the holoprotein did not correlate simply with helical propensities in the disordered regions. PMID- 18398854 TI - Structural propensities in the heme binding region of apocytochrome b5. II. Heme conjugates. AB - In the absence of heme cofactor, the water-soluble domain of rat microsomal cytochrome b5 (cyt b5) contains a long flexible region within its 42-residue heme binding loop. Heme capture induces this region to fold into a well-defined structure containing helices H3-H5, each separated by a turn, with His39 and His63 serving as axial ligands to the heme iron. We have shown that the H4 region of the apoprotein has the greatest tendency for disorder within the isolated binding loop. Here, the effect of the His63-iron bond and proximity of heme plane on the population of helical conformation in H4 and H5 was investigated by synthesis and characterization of a peptide-sandwiched mesoheme construct in which two H4-H5 peptides were covalently attached to a single cofactor. Spectroscopic data indicated that a holoprotein-like bis-histidine coordination state was achieved over a pH range from 7 to 9. Trifluoroethanol titrations of the construct and the analogous free peptide under these pH conditions revealed that heme proximity and iron ligation were insufficient to promote helix formation in H4 and H5. These observations were used to assess the role of disordered regions in heme capture and the loop-scaffold interface in holoprotein folding and stability. PMID- 18398855 TI - Schinzel-Giedion syndrome: report of splenopancreatic fusion and proposed diagnostic criteria. AB - We report on the 46th patient with Schinzel-Giedion syndrome (SGS) and the first observation of splenopancreatic fusion in this syndrome. In the antenatal period, a male fetus was found to have bilateral hydronephrosis. Postnatally, in keeping with a diagnosis of SGS, there were large fontanelles, ocular hypertelorism, a wide, broad forehead, midface retraction, a short, upturned nose, macroglossia, and a short neck. Other anomalies included cardiac defects, widened and dense long bone cortices, cerebral ventriculomegaly, and abnormal fundi. Splenopancreatic fusion, usually encountered in trisomy 13, was found on autopsy. Schinzel-Giedion syndrome is likely a monogenic condition for which neither the heritability pattern nor pathogenesis has yet been determined. A clinical diagnosis may be made by identifying the facial phenotype, including prominent forehead, midface retraction, and short, upturned nose, plus one of either of the two other major distinguishing features: typical skeletal abnormalities or hydronephrosis. Typical skeletal anomalies include a sclerotic skull base, wide supraoccipital-exoccipital synchondrosis, increased cortical density or thickness, and broad ribs. Other highly supportive features include neuroepithelial tumors (found in 17%), hypertrichosis, and brain abnormalities. Severe developmental delay and poor survival are constant features in reported patients. PMID- 18398856 TI - Heterocyclic carbenes: synthesis and coordination chemistry. AB - The chemistry of heterocyclic carbenes has experienced a rapid development over the last years. In addition to the imidazolin-2-ylidenes, a large number of cyclic diaminocarbenes with different ring sizes have been described. Aside from diaminocarbenes, P-heterocyclic carbenes, and derivatives with only one, or even no heteroatom within the carbene ring are known. New methods for the synthesis of complexes with N-heterocyclic carbene ligands such as the oxidative addition or the metal atom template controlled cyclization of beta-functionalized isocyanides have been developed recently. This review summarizes the new developments regarding the synthesis of N-heterocyclic carbenes and their metal complexes. PMID- 18398862 TI - Enrichment of eicosapentaenoic acid and docosahexaenoic acid from sardine by products by supercritical fluid fractionation. AB - Based on the principle of supercritical chromatography, a fractionation process for fatty acid methyl esters (FAMEs) under supercritical conditions was studied with the aim of obtaining in extracts highly enriched eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish cannery waste. Yields of the fractionation process were enhanced by using adjuvant material and modifying CO(2) volumetric density and temperature. The combination of adjuvant properties with optimized pressure and temperature conditions permitted efficient fractionations of FAMEs, according to chain length and degree of unsaturation, in high purity (up to 95% of FAME) and good yields (45% of EPA and DHA starting potential). This work shows how green technologies, such as supercritical processes, can constitute a good alternative to the use of organic solvents in classical methods for valorisation of complex food industry waste. PMID- 18398863 TI - Stability-indicating assay of sodium cromoglicate in ophthalmic solution using mixed-mode hydrophilic interaction chromatography. AB - A hydrophilic interaction chromatographic (HILIC) procedure for the quantification of Sodium Cromoglicate (SCG) in ophthalmic solution is developed. Mobile phase consists of ACN and buffer, 86:14 v/v. Atlantis HILIC-Si column, 25 cm x 4.6 mm, is used as stationary phase. Detection is carried out using a variable wavelength UV-Vis detector at 326 nm. Linearity range and percent recoveries for SCG were 50-400 mug/mL and 100.44%, respectively. The SCG HILIC-UV assay was validated according to the International Conference on Harmonization guidelines. The method separates two impurities and degradation products resulting from stress environment. Influence of organic solvent, ionic strength and mobile phase pH on the retention of SCG is studied. The paper provides optimization of polar anionic solute (SCG) on unmodified silica by HILIC. Proposed method can be used as a stability-indicating assay for SGC and can be proved to be beneficial in ESI-MS for enhanced sensitivity. PMID- 18398864 TI - Supercritical fluid extraction of amino acids from birch (Betula pendula Roth) leaves and their liquid chromatographic determination with fluorimetric detection. AB - A method for supercritical fluid extraction (SFE) of amino acids was adapted and optimal experimental conditions were selected for a matrix consisting of dry leaves. The matrix-dependent SFE method uses a mixture of MeOH-H(2)O-acetonitrile (10:10:1 v/v/v) as a modifier (0.5 mL in situ, 300 muL on-line) at 70 degrees C and 40 MPa and no HCl is needed as an entrainer. The amino acids were quantified using high-performance liquid chromatography with fluorimetric detection (HPLC/FLD) after gradient elution on Zorbax Eclipse AAA columns (4.6x150 mm, 3.5 mum) with aqueous Na(2)HPO(4 )buffer of pH 7.8 and ACN-MeOH-water as a mobile phase. In comparison with Soxhlet extraction, SFE gave higher recovery and selectivity, but it required longer extraction time (90 min) and it was more labor-intensive (clean-up step after the pre-concentration). Both methods should be used separately or in combination according to the matrix, number of samples, and levels of ballast compounds. PMID- 18398865 TI - Supercritical fluid extraction of polyhalogenated pollutants from aquaculture and marine environmental samples: a review. AB - This article focuses on the state-of-the-art in sample preparation using supercritical fluid extraction (SFE), to monitor the content of polyhalogenated pollutants in aquaculture and marine environmental samples. Marine sediments and biological applications, including several types of samples matrices (fish, shellfish, seaweed and fish feed) and analyte groups (polychlorinated biphenyls (PCBs), polybrominated biphenyls (PBBs), polybrominated diphenylethers (PBDEs), polychlorinated dibenzo-p-dioxin (PCDD)/Fs and organochlorinated pesticide (OCPs)) are discussed with respect to SFE use and optimisation of conditions. We also discuss the great analytical potential of SFE, the integration of the extraction and clean-up steps for rapid sample processing justifying its use for routine work. The most recent SFE applications to the determination of these pollutants in marine environmental (biota and sediment) samples, published in the last 15 years, are reviewed. PMID- 18398866 TI - Overview of the retention in subcritical fluid chromatography with varied polarity stationary phases. AB - Retention and separation of achiral compounds in supercritical fluid chromatography (SFC) depend on numerous parameters: some of these parameters are identical to those encountered in HPLC, such as the mobile phase polarity, while others are specific to SFC, as the density changes of the fluid, due to temperature and/or pressure variations. Additional effects are also related to the fluid compressibility, leading to unusual retention changes in SFC, for instance when flow rate or column length is varied. These additional effects can be minimised by working at lower temperatures in the subcritical domain, simplifying the understanding of retention behaviours. In these subcritical conditions, varied modifiers can be mixed to carbon dioxide, from hexane to methanol, allowing tuning the mobile phase polarity. With nonpolar modifiers, polar stationary phases are classically used. These chromatographic conditions are close to the ones of normal-phase LC. The addition of polar modifiers such as methanol or ACN increases the mobile phase polarity, allowing working with less polar stationary phases. In this case, despite the absence of water, retention behaviours generally follow the rules of RP LC. Moreover, because identical mobile phases can be used with all stationary phase types, from polar silica to nonpolar C18-bonded silica, the classical domains, RP and normal-phase, are easily brought together in SFC. A unified classification method based on the solvation parameter model is proposed to compare the stationary phase properties used with the same subcritical mobile phase. PMID- 18398867 TI - Enterolactone restricts the proliferation of the LNCaP human prostate cancer cell line in vitro. AB - Ecological data suggest a long-term diet high in plant material rich in biologically active compounds, such as the lignans, can significantly influence the development of prostate cancer over the lifetime of an individual. The capacity of a pure mammalian lignan, enterolactone (ENL), to influence the proliferation of the LNCaP human prostate cancer cell line was investigated as a function of cell density, metabolic activity, expression and secretion of prostate specific antigen (PSA), cell cycle profile, and the expression of genes involved in development and progression of prostate cancer. Treatment with a subcytotoxic concentration of ENL (60 muM for 72 h) was found to reduce: cell density (57.5%, SD 7.23, p < 0.001), metabolic activity (55%, SD 0.03, p < 0.001), secretion of PSA (48.50% SD 4.74, p = 0.05) and induce apoptosis (8.33 fold SD 0.04, p = 0.001) compared to untreated cells. Cotreatment with 10 muM etoposide was found to increase apoptosis by 50.17% (SD 0.02, p < 0.001). Additionally, several key genes (e. g. MCMs, survivin and CDKs) were beneficially regulated by ENL treatment (p < 0.05). The data suggest that the antiproliferative activity of ENL is a consequence of altered expression of cell cycle associated genes and provides novel molecular evidence for the antiproliferative properties of a pure lignan in prostate cancer. PMID- 18398868 TI - DGGE and real-time PCR analysis of lactic acid bacteria in bacterial communities of the phyllosphere of lettuce. AB - Food associated indigenous microbial communities exert antagonistic effects on pathogens and may routinely deliver health relevant microorganisms to the GI tract. By using molecular, culture independent methods including PCR-DGGE of 16S rDNA-coding regions and real-time PCR (RT-PCR) as well as BIOLOG metabolic fingerprinting, microbial communities on lettuce were analyzed in samples from fields, from supermarkets and soil. Amplified 16S rRNA gene sequences (57.7%) could be assigned to species previously reported as typical for the phyllosphere including Pantoea agglomerans, Pseudomonas flavescens, Moraxella spp., and Mycobacterium spp. 71.8% of the sequences obtained represented so far undescribed taxa. Principal component analysis of BIOLOG metabolic profiles indicated a seasonal variation in the lettuce phyllosphere microbial community structure. Various lactic acid bacteria were detected including several Lactobacillus and Leuconostoc species in particular on lettuce from organic farming. By RT-PCR lactobacilli were found with a range of abundances from 1x10(4 )to 1x10(5 )copies/g lettuce. Considering the importance of salad in many diets lettuce may contribute to a constant supply with LAB. PMID- 18398869 TI - Effect of curcumin supplementation on blood glucose, plasma insulin, and glucose homeostasis related enzyme activities in diabetic db/db mice. AB - We investigated the effect of curcumin on insulin resistance and glucose homeostasis in male C57BL/KsJ-db/db mice and their age-matched lean non-diabetic db/+ mice. Both db/+ and db/db mice were fed with or without curcumin (0.02%, wt/wt) for 6 wks. Curcumin significantly lowered blood glucose and HbA 1c levels, and it suppressed body weight loss in db/db mice. Curcumin improved homeostasis model assessment of insulin resistance and glucose tolerance, and elevated the plasma insulin level in db/db mice. Hepatic glucokinase activity was significantly higher in the curcumin-supplemented db/db group than in the db/db group, whereas glucose-6-phosphatase and phosphoenolpyruvate carboxykinase activities were significantly lower. In db/db mice, curcumin significantly lowered the hepatic activities of fatty acid synthase, beta-oxidation, 3-hydroxy 3-methylglutaryl coenzyme reductase, and acyl-CoA: cholesterol acyltransferase. Curcumin significantly lowered plasma free fatty acid, cholesterol, and triglyceride concentrations and increased the hepatic glycogen and skeletal muscle lipoprotein lipase in db/db mice. Curcumin normalized erythrocyte and hepatic antioxidant enzyme activities (superoxide dismutase, catalase, gluthathione peroxidase) in db/db mice that resulted in a significant reduction in lipid peroxidation. However, curcumin showed no effect on the blood glucose, plasma insulin, and glucose regulating enzyme activities in db/+ mice. These results suggest that curcumin seemed to be a potential glucose-lowering agent and antioxidant in type 2 diabetic db/db mice, but had no affect in non-diabetic db/+ mice. PMID- 18398870 TI - Immunomodulatory effects of curcumin in allergy. AB - Recent years have witnessed a global increase in allergy and asthma, particularly in developed countries. Attempts to develop effective control measures for allergy and asthma resulted in the exploration of alternate medicines including herbal remedies traditionally used in old world countries. Turmeric is known for its multiple health restoring properties, and has been used in treating several diseases including several respiratory disorders. Turmeric is a common spice used in the culinary preparations in South and East Asian countries. The active component of turmeric is curcumin, a polyphenolic phytochemical, with anti inflammatory, antiamyloid, antiseptic, antitumor, and antioxidative properties. Curcumin was reported to have antiallergic properties with inhibitory effect on histamine release from mast cells. The effectiveness of curcumin in allergy and asthma has been further investigated using a murine model of allergy. The results indicate a marked inhibition of allergic response in animals treated with curcumin suggesting a major role for curcumin in reducing the allergic response. The present review focuses on the results of research aimed to understand the immunomodulation induced by curcumin and its associated roles in the amelioration of allergy. These findings needed further evaluation, extrapolation, and confirmation before using curcumin for controlling allergy and asthma in humans. PMID- 18398871 TI - Fractionation of polyphenol-enriched apple juice extracts to identify constituents with cancer chemopreventive potential. AB - Apples and apple juices are widely consumed and rich sources of phytochemicals. The aim of the present study was to determine which apple constituents contribute to potential chemopreventive activities, using a bioactivity-directed approach. A polyphenol-enriched apple juice extract was fractionated by various techniques. Extract and fractions were tested in a series of test systems indicative of cancer preventive potential. These test systems measured antioxidant effects, modulation of carcinogen metabolism, anti-inflammatory and antihormonal activities, and antiproliferative potential. Regression analyses indicated that 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging potential correlated with the sum of low molecular weight (LMW) antioxidants (including chlorogenic acid, flavan-3-ols, and flavonols) and procyanidins, whereas peroxyl radicals were more effectively scavenged by LMW compounds than by procyanidins. Quercetin aglycone was identified as a potent Cyp1A inhibitor, whereas phloretin and (-)-epicatechin were the most potent cyclooxygenase 1 (Cox-1) inhibitors. Aromatase and Cyp1A inhibitory potential and cytotoxicity toward HCT116 colon cancer cells increased with increasing content in procyanidins. Overall, apple juice constituents belonging to different structural classes have distinct profiles of biological activity in these in vitro test systems. Since carcinogenesis is a complex process, combination of compounds with complementary activities may lead to enhanced preventive effects. PMID- 18398872 TI - Resveratrol inhibits migration and invasion of human breast-cancer cells. AB - Metastasis is the primary cause of death from breast cancer. Cell migration and invasion play important roles in neoplastic metastasis. The insulin-like growth factor (IGF-1) stimulates cell migration through activation of PI-3K/Akt signaling pathway. IGF-1 induces the tumorigenicity of many types of cancer cells and is critical for metastatic cell spread in estrogen receptor (ER)-negative breast-cancer cells. Matrix metalloproteinase-2 (MMP-2) is a key enzyme in the degradation of extracellular matrices and its expression has been dysregulated in breast cancer invasion and metastasis. Resveratrol exhibited potential anticarcinogenic activities in several studies. However, the inhibitory effects of resveratrol on the expression of MMP-2, migration and invasion of breast cancer cell have not been demonstrated yet. In the present study, we investigated the anti-invasive mechanism of resveratrol in human breast cancer MDA-MB 435cells. Here, we showed that IGF-1 is a potent stimulant of the migration of ER negative human breast-cancer cells. Resveratrol could inhibit IGF-1-mediated cell migration of MDA-MB 435 in vitro. The inhibitory effect of resveratrol was mediated in part through the suppression of the activation of PI-3K/Akt signaling pathway. Furthermore, IGF-1-mediated expression of MMP-2 was significantly inhibited by resveratrol in concomitance with alteration of cell invasion. PMID- 18398873 TI - Exploring sequence space: profile analysis and protein-ligand docking to screen omega-aminotransferases with expanded substrate specificity. AB - omega-Aminotransferase (omegaAT) is an interesting biocatalyst for the preparation of chiral amines, which are widely used as building blocks for pharmaceuticals, agrochemicals and fine chemicals. With the assumption that substrate and sequence spaces are in the process of co-evolution, we explored sequences space to screen the enzymes showing activity to new target compounds. Bacterial genome sequences (n=527) were analyzed by the profiles of subgroups in aminotransferase group II including ornithine aminotransferase (orAT), acetylornithine aminotransferase (aoAT), omegaAT, gamma-aminobutyrate aminotransferase (GABAAT) and 7,8-diaminopelargonate aminotransferase (DAPAAT). We selected the sequences having a Z score of 0-1.8 to guarantee the omegaAT reaction and to avoid the typical omegaAT sequences. Among the selected sequences, we filtered out the sequences with very low Z scores for the rest of four subgroups in aminotransferase group II to consider the diversity. For the selected sequences, we performed protein-ligand docking simulations to predict the docking pose of amino acceptor. Throughout all the analysis procedures, several candidate aminotransferase sequences for the asymmetric synthesis of chiral amines were obtained. An efficient procedure for virtual screening of novel enzymes was demonstrated. PMID- 18398874 TI - The whereabouts of transmembrane proteins from rat brain synaptosomes during two dimensional gel electrophoresis. AB - Little is known about what happens to transmembrane proteins (TMP) in 2-DE. In order to obtain more insight into the whereabouts of these proteins we prepared membrane-enriched synaptosomes from rat frontal cortex and washed them with 7 M urea or Na(2)CO(3). From each preparation, 200 microg protein was loaded on 2-DE gels covering the 4-7 and 6-11 pH ranges, respectively. MALDI-MS/MS analysis detected only 3 TMP among 421 identified spots. However, when the samples had been washed with Na(2)CO(3), only few well-focused spots remained detectable on the gel covering the pH 6-11 range. Instead, a heavily ruthenium-stained smear became visible at the upper edge of the gel at the location where the samples had been applied by cup loading. LC-MS/MS analysis revealed that this smear contained 38 unfocused TMP with up to 12 transmembrane helices. After transfer to the second dimension, no major areas of protein staining were left on the IPG strips. This indicates that after extraction and denaturation the TMP may form high molecular aggregates, due to their "hydrophobic interactions". These aggregates enter the IPG strips, but do not focus regularly. They are then transferred onto the 2-DE-gels, where they remain caught at the upper edge. PMID- 18398875 TI - Proteomics analysis of differential expression of cellular proteins in response to avian H9N2 virus infection in human cells. AB - We present the first proteomic analysis on the cellular responses to avian influenza virus (H9N2) infection in a human cell line in different time courses in order to search for target proteins for viral pathogenesis/adaptation studies. By using 2-DE coupled with MALDI-TOF MS and nano-ESI-MS/MS, we identified a set of differentially expressed cellular proteins, including cytoplasmic actin, cytokeratin, prohibitin, enoyl-CoA hydratase, peptide-prolyl cis-trans isomerase A (PPIase A), chloride intracellular channel protein 1, pyruvate dehydrogenase E1 component subunit beta, adenine phosphoribosyltransferase, guanine nucleotide binding protein subunit beta, nucleoside diphosphate kinase A, elongation factor 1-beta and splicing factor, arginine/serine rich 1. The most significant changes in different time courses were found in cytoplasmic actin and cytokeratin, both of which constituted the major components of cytoskeleton network in the cells. The obtained data suggested a possible role of the cytoskeleton during avian influenza virus infection of mammalian cells, which might help for better understanding of the dynamics of avian influenza virus and host interaction in mammalian cell setting. PMID- 18398876 TI - A highly uniform UV transillumination imaging system for quantitative analysis of nucleic acids and proteins. AB - The digital fluorescent imaging for documentation and analysis of gel electrophoretic separations of nucleic acids and proteins is widely used in quantitative biology. Most fluorescent stains used in postelectrophoretic analysis of proteins and nucleic acids have significant excitation peaks with UV light (300-365 nm), making midrange UV (UV-B) as the excitation source of choice. However, coupling quantitative CCD imaging with UV is difficult due to lack of uniformity found in typical UV transilluminators. The apparent amount of those macromolecules depends on the position of the gel band on the imaging surface of the transilluminator. Here, we report the development and validation of a highly uniform UV transillumination system. Using a novel high density lighting system containing a single lamp formed into a high density grid, an electronic ballast, a phosphor coating, and a bandpass filter to convert 254 nm light produced to 300 340 nm, uniformity of 80% CV observed in typical UV transilluminators. This system has been used for the quantitative analysis of electrophoretically separated nucleic acids and proteins (CV10(8)-fold increase in affinity compared with the mono P(k) ligand. This intrinsic high-affinity AuNP probe specifically captured the recombinant B-Slt from Escherichia coli lysate, and the resulting purity of the B-Slt was >95 %. We also developed a robust P(k)-AuNP-based detection method for Slt-I by combining the technique with silver enhancement. PMID- 18398882 TI - A simple method for preparing peptide C-terminal thioacids and their application in sequential chemoenzymatic ligation. PMID- 18398883 TI - Boolean logic gates that use enzymes as input signals. AB - Biochemical systems that demonstrate the Boolean logic operations AND, OR, XOR, and InhibA were developed by using soluble compounds, which represent the chemical "devices", and the enzymes glucose oxidase (GOx), glucose dehydrogenase (GDH), alcohol dehydrogenase (AlcDH), and microperoxidase-11 (MP-11), which operated as the input signals that activated the logic gates. The enzymes were used as soluble materials and as immobilized biocatalysts. The studied systems are proposed to be a step towards the construction of "smart" signal-responsive materials with built-in Boolean logic. PMID- 18398884 TI - GeB4O9 x H2en: an organically templated borogermanate with large 12-ring channels built by B4O9 polyanions and GeO4 units: host-guest symmetry and charge matching in triangular-tetrahedral frameworks. AB - A new borogermanate open-framework, FJ-18, with intersecting 12-/8-, 9-, and 9 ring channels in multidimensions was synthesized by using an organic diamine as structure-directing agent (SDA). X-ray diffraction showed that its structure is constructed by flexible connection of strictly alternate B(4)O(9) clusters (with fictitious tetrahedral geometry) and GeO(4) tetrahedra to form a zeotype framework with the CrB(4) topology. The topology and structural relations between FJ-18 and FJ-16, a known borogermanate based on the linkage of B(4)O(9) clusters and GeO(4) units, are described in detail. Different guest SDAs can, by means of their shape, size, and charge, directly influence the structure of the host framework, and the origin of these phenomena can be attributed to fundamental parameters such as host-guest symmetry and charge matching through H-bonding interactions, the concept of which has for the first time been extended to tetrahedral-triangular frameworks. This result is a part of our ongoing work aimed at making new large-pore materials constructed from B-O polyanions and Ge-O cluster units. PMID- 18398885 TI - Oxidation of aromatic alcohols in irradiated aqueous suspensions of commercial and home-prepared rutile TiO(2): a selectivity study. AB - The photocatalytic oxidation of benzyl alcohol (BA) and 4-methoxybenzyl alcohol (MBA) has been performed in pure water by using commercial TiO(2) samples (Sigma Aldrich, Merck, Degussa P25) and rutile TiO(2) prepared from TiCl(4) at low temperature. Particular attention has been devoted to the identification of the produced aromatic compounds along with the formed CO(2). Oxidation products such as the corresponding aromatic aldehyde and acid, as well as mono- and dihydroxylated aldehydes have been detected. The home-prepared rutile sample showed a marked selectivity towards the formation of the aromatic aldehyde (38 and 60 % for BA and MBA, respectively), resulting in a three- to sevenfold improvement relative to commercial samples, with the only byproduct being CO(2). This catalyst was found to be the most selective in the formation of aldehyde in water. By using the commercial or the calcined home-prepared samples, many hydroxylated aromatic compounds were detected besides the aldehyde and the acid. This finding points to a higher selectivity performance of the home-prepared rutile relative to the commercial TiO(2) samples. Some of the home-prepared samples were also dialysed to check the influence of the presence of Cl(-) species on catalyst reactivity and selectivity. We have attempted to explain the different reaction rate and selectivity observed for MBA and BA. PMID- 18398886 TI - Hierarchically self-assembled host-guest network at the solid-liquid interface for single-molecule manipulation. PMID- 18398887 TI - The influence of guanine on DNA hole transport efficiency. PMID- 18398888 TI - A transient V(III)-alkylidene complex: oxidation chemistry including the activation of N2 to afford a highly porous honeycomb-like framework. PMID- 18398889 TI - Mechanism-guided improvements to the single molecule oxidation of carbon nanotube sidewalls. AB - Real-time monitoring of carbon nanotube conductance during electrochemical and chemical etching reveals the electronic signatures of individual bond alteration events on the nanotube sidewall. Tracking the conductance of multiple single molecule experiments through different synthetic protocols supports putative mechanisms for sidewall derivatization. Insights gained from these mechanistic observations imply the formation of sidewall carboxylates, which are useful as handles for bioconjugation. We describe an electronic state required for efficacious chemical treatment. Such real-time monitoring can improve carboxylate yields to 45 % or more. The experiments illustrate the power of molecular nanocircuits to uncover and direct the mechanisms of chemical reactions. PMID- 18398890 TI - Dynamics of the size distribution of CdTe quantum dot ensembles during growth in liquid and crystalline phases. AB - We recently reported that the growth rate of colloidal CdTe nanoparticles decreases by orders of magnitude when the particles undergo a phase transition from liquid to crystal. The dynamics of nanoparticle growth are dominated by this factor rather than the size dependence of the chemical potential. Herein we discuss how the phase transition affects the size distribution and photoluminescence quantum efficiency of the nanoparticles. We suggest that the absorption linewidth is a better monitor of size distribution than the photoluminescence linewidth because the photoluminescence quantum efficiency, which affects the latter via energy transfer, varies substantially with reaction time. We find that the size distribution broadens in the early stages of growth possibly because of inhomogeneities in the phase transition radius or because particles nucleated at later times coalesce with nanocrystals. The quantum efficiency is enhanced when tellurium is depleted in the reaction solution, giving a cadmium-enriched surface. Batches with high initial tellurium and cadmium concentrations show a substantial amount of delayed nucleation, lower quantum efficiency and some anisotropic growth. PMID- 18398891 TI - Three-residue turns in alpha/beta-peptides and their application in the design of tertiary structures. AB - A new three-residue turn was serendipitously discovered in alpha/beta hybrid peptides derived from alternating C-linked carbo-beta-amino acids (beta-Caa) and L-Ala residues. The three-residue beta-alpha-beta turn at the C termini, nucleated by a helix at the N termini, resulted in helix-turn (HT) supersecondary structures in these peptides. The turn in the HT motif is stabilized by two H bonds-CO(i-2)-NH(i), with a seven-membered pseudoring (gamma turn) in the backward direction, and NH(i-2)-CO(i), with a 13-membered pseudoring in the forward direction (i being the last residue)--at the C termini. The study was extended to generalize the new three-residue turn (beta-alpha-beta) by using different alpha- and beta-amino acids. Furthermore, the HT motifs were efficiently converted, by an extension with helical oligomers at the C termini, into peptides with novel helix-turn-helix (HTH) tertiary structures. However, this resulted in the destabilization of the beta-alpha-beta turn with the concomitant nucleation of another three-residue turn, alpha-beta-beta, which is stabilized by 11- and 15-membered bifurcated H bonds. Extensive NMR spectroscopic studies were carried out to delineate the secondary and tertiary structures in these peptides, which are further supported by molecular dynamics (MD) investigations. PMID- 18398892 TI - 1,2,3-Triazole CH...Cl(-) contacts guide anion binding and concomitant folding in 1,4-diaryl triazole oligomers. PMID- 18398893 TI - Episodic infliximab treatment induces infusion reactions. PMID- 18398894 TI - Clinical role of calprotectin assay in determining histological relapses in children affected by inflammatory bowel diseases. AB - BACKGROUND: Inflammatory bowel diseases (IBD) are characterized by periods of remission with recurrent episodes of symptom exacerbation because of acute intestinal inflammation, which is correctly evaluated by endoscopy with biopsy sampling. However, many surrogate markers of intestinal inflammation, including fecal calprotectin (FC), are detected as potential predictors of mucosal inflammation in IBD patients. The aim of our study was to retrospectively assess the clinical efficacy of the calprotectin assay in determining histological relapses of pediatric IBD patients. METHODS: We retrospectively reviewed the histological examinations, clinical records, and FC values of patients who had undergone colonoscopy at our hospital over an 8-year period, from December 31, 1998, to December 31, 2006. Only patients with a first histological examination showing a quiescent IBD who submitted to a second histological examination during the next 3 years were selected. RESULTS: Seventy-three IBD patients, all with a first biopsy showing a quiescent IBD, were studied; at the second histological examination, 32 presented with relapse and 41 presented with remission. Relapsed patients showed significantly increased FC levels compared with nonrelapsed patients. A FC value of 275 mug/g achieved sensitivity and negative predictive value of 97% and specificity and positive predictive value of 85% in predicting histological relapse. CONCLUSIONS: FC seems to be a direct measure of intestinal inflammation and therefore a good marker of the risk of histological relapse in pediatric IBD patients. The application of this test in clinical practice may enable the avoidance of invasive tests as well as targeting treatment. PMID- 18398895 TI - JAK2 V617F mutation is not involved in thromboembolism in IBD. PMID- 18398896 TI - Oral contrast-enhanced sonography for the diagnosis and grading of postsurgical recurrence of Crohn's disease. AB - BACKGROUND: Postsurgical recurrence (PSR) is very common in patients with Crohn's disease (CD) and previous surgery. Endoscopy is crucial for the diagnosis of PSR, also showing high prognostic value. Bowel sonography (BS) with or without oral contrast enhancement (OCBS) is accurate for CD diagnosis but its role in PSR detection and grading is poorly investigated. The aim was to evaluate the diagnostic accuracy of BS and OCBS for PSR compared to the endoscopical Rutgeerts's grading system. METHODS: We prospectively performed endoscopy, BS, and OCBS in 40 CD patients with previous bowel resection to provide evidence of possible PSR. Endoscopy, BS, and OCBS were executed 1 year after surgery, with PSR diagnosis and grading made in accordance with Rutgeerts. BS and OCBS were considered suggestive for PSR in the presence of bowel wall thickness (BWT) >3 mm. OCBS was performed after ingestion of 750 mL of polyethylene glycol (PEG). Also, a receiver operating characteristic (ROC) curve was constructed in order to define the best cutoff of BWT to discriminate mild from severe PSR (grade 0-2 versus 3-4 of Rutgeerts) for both BS and OCBS. RESULTS: In all, 22 out of the 40 CD showed an endoscopic evidence of PSR (55%). A severe PSR was present in 14 patients (64%). Sensitivity, specificity, and positive and negative predictive values were 77%, 94%, 93%, and 80% for BS, and 82%, 94%, 93%, and 84% for OCBS. On the ROC curve a BWT >5 mm showed sensitivity, specificity, and positive and negative predictive values of 93%, 96%, 88%, and 97% for the diagnosis of severe PSR at BS, while a BWT >4 mm was the best cutoff differentiating the mild from the severe CD recurrence for OCBS, with a sensitivity, specificity, and positive and negative predictive values of 86%, 96%, 97%, and 79%, respectively. CONCLUSIONS: Both BS and OCBS show good sensitivity and high specificity for the diagnosis of PSR in CD, with a BWT >5 mm for BS and BWT >4 mm for OCBS strongly indicative of severe endoscopic PSR. Accordingly, these techniques could replace endoscopy for the diagnosis and grading of PSR in many cases. PMID- 18398897 TI - Diagnostic medical radiation in IBD. PMID- 18398898 TI - Immunosuppressant supplementation to infliximab in Crohn's disease with extraintestinal involvement: a benefit in the long term? PMID- 18398899 TI - Blind J limb syndrome: recurrent enterocutaneous fistulae from an ileal pouch. PMID- 18398900 TI - Ganoderma applanatum: a promising mushroom for antitumor and immunomodulating activity. AB - The antitumor effect of exo-biopolymer (EXP) produced by Ganoderma applanatum was investigated using sarcoma-180 bearing mice. EXP, when administered (10-80 mg/kg body weight: BW) intraperitoneally, significantly inhibited the growth of solid tumor and increased the natural killer (NK) cell activity. A dose of 40 mg/kg BW was found to be highly effective, as it reduced the tumor formation by 39.7%, and increased the NK cell activity of splenocytes by 51.6% compared with the control group. The complement activity of EXP was increased in accordance with an increase in concentration. The phosphatase activity of macrophages was increased by 0.7-fold (200 microg/mL) compared with the control group. This EXP contained 58.9% carbohydrate and 17.1% protein. The major sugar of EXP was composed of mannose and glucose, while the protein mainly consisted of serine, glycine and aspartic acid. PMID- 18398901 TI - Influence of particle size and antigen binding on effectiveness of aluminum salt adjuvants in a model lysozyme vaccine. AB - It has been suggested that agglomeration of aluminum salt adjuvant particles during freezing and drying can cause loss of immunogenicity of vaccines formulated with such adjuvants. In this study, we tested this hypothesis and examined the immune response in a murine model to various liquid, freeze-thawed, and lyophilized vaccine formulations, using lysozyme as a model antigen. The various processing techniques and excipient levels resulted in a wide range of particle size distributions (PSDs) and antigen-adjuvant binding levels. Anti lysozyme titers were independent of the PSD for vaccines adjuvanted with either aluminum hydroxide or aluminum phosphate and also were unaffected by the level of antigen binding to the adjuvant. PMID- 18398902 TI - Crystal chemistry of sibutramine: thermal, diffractometric and spectroscopic characterization. AB - Monohydrated sibutramine hydrochloride is a widely used active ingredient for the treatment of obesity. An anhydrous form of sibutramine hydrochloride was prepared starting from its monohydrate form upon heating it at 140 degrees C for 15 min. This dehydration process was monitored using conventional TG/DSC methods. Heated above 190 degrees C, sibutramine hydrochloride sublimes and recrystallizes on the cold walls of the test tube, giving platelet shaped crystals suitable for single crystal X-ray diffraction analysis: monoclinic, P2(1)/n, a = 7.321(2) A, b = 25.456(5) A, c = 9.750(3) A, beta = 101.60(2) degrees , V = 1779.9(8) A(3), Z = 4. At variance, sibutramine free base was typically recovered as a viscous oily material, upon treatment of its hydrochloride salt in ethyl acetate solution. Recrystallization from hexane yielded a white polycrystalline powder, the structure of which was determined by unconventional ab initio X-ray powder diffraction analysis: triclinic, P-1, a = 8.6578(3) A, b = 9.3318(3) A, c = 11.1224(4) A, alpha = 110.434(3) degrees , beta = 100.159(3) degrees , gamma = 89.201(2) degrees , V = 827.76(5) A(3), Z = 2. Sibutramine, in its different crystalline environments, was also fully characterized by solid state (13)C NMR analyses. Additional spectral information was obtained by collecting spectra of a metastable, oily sample, before it slowly recrystallizes (within hours). PMID- 18398903 TI - Cancer phytotherapeutics: role for flavonoids at the cellular level. AB - Dietary foods and fruits possess an array of flavonoids with unique chemical structure and diverse bioactivities relevant to cancer. Numerous epidemiological studies have validated the inverse relation between the consumption of flavonoids and the risk of cancer. Flavonoids possess cancer blocking and suppressing effects. Flavonoids modulate various CYPs involved in carcinogen activation and scavenging reactive species formed from carcinogens by CYP-mediated reactions. They induce biosynthesis of several CYPs. They are involved in the regulation of enzymes of phase-II responsible for xenobiotic biotransformation and colon microflora. Since cytochromes P450, P-gp and phase-II enzymes are involved in the metabolism of drugs and in the processes of chemical carcinogenesis, interactions of flavonoids with these systems hold great promise for their therapeutic potential. The role of flavonoids also includes the inhibition of activation of pro-carcinogens, inhibition of proliferation of cancer cells, selective death of cancer cells by apoptosis, inhibition of metastasis and angiogenesis, activation of immune response against cancer cells, modulation of the inflammatory cascade and the modulation of drug resistance. This has greatly extended the goal of cancer therapy from eradicating the affected cells to control of the cancer phenotype. Phytotherapy is being used in combination with other therapies as phytonutrients have been shown to work by nutrient synergy. PMID- 18398904 TI - Bioassay-guided purification and identification of PPARalpha/gamma agonists from Chlorella sorokiniana. AB - This study isolated agonists of peroxisome proliferator activated receptors (PPARs) from the green algae Chlorella sorokiniana, using a bioassay-guided purification strategy. PPARs are widely recognized as the molecular drug targets for many diseases including hyperglycemia, diabetes, obesity and cancer. Two independent bioassays were developed. The first is the scintillation proximity assay, a ligand binding assay. The other is the cell-based transcriptional activation assay which uses the Dual-Luciferase reporter system as the reporter gene under the control of the PPAR response element. Using these two assays, a PPARgamma-active fraction, CE 3-3, was obtained from C. sorokiniana extracts, which was also able to activate PPARalphamediated gene expression. To elucidate the active ingredients in the CE 3-3 fraction, GC-MS analysis was employed. The results showed that the CE 3-3 fraction consisted of at least ten fatty acids (FAs). The bioactivities of several of the individual FAs were evaluated for their PPARgamma activity and the results showed that linolenic acid and linoleic acid were the most potent FAs tested. Our studies indicate that Chlorella sorokiniana could have potential health benefits through the dual activation of PPARalpha/gamma via its unique FA constituents. PMID- 18398905 TI - Comparative molecular dynamics simulations of histone deacetylase-like protein: binding modes and free energy analysis to hydroxamic acid inhibitors. AB - Histone deacetylases (HDACs) play an important role in gene transcription, and inhibitors of HDACs can induce cell differentiation and suppress cell proliferation in tumor cells. Histone deacetylase1 (HDAC1) binds suberanilohydroxamic acid (SAHA) and 7-phenyl-2, 4, 6-hepta-trienoyl hydroxamic acid (CG-1521) with moderately low affinity (DeltaG = -8.6 and -7.8 kcal mol( 1)). The structurally related (E)-2-(3-(3-(hydroxyamino)-3-oxoprop-1-enyl)phenyl) N(1),N(3)-diphenylmalonamide (SK-683), a Trichostatin A (TSA)-like HDAC1 inhibitor, and TSA are bound to the HDAC1 with -12.3 and -10.3 kcal mol(-1) of DeltaG, higher binding free energies than SAHA and CG-1521. Histone deacetylase like protein (HDLP), an HDAC homologue, shows a 35.2% sequence identity of HDLP and human HDAC1. Molecular dynamics simulation and the molecular mechanics/generalized-Born surface area (MM-GBSA) free energy calculations were applied to investigate the factors responsible for the relatively activity of these four inhibitors to HDLP. In addition, computational alanine scanning of the binding site residues was carried out to determine the contribution components from van der Waals, electrostatic interaction, nonpolar and polar energy of solvation as well as the effects of backbones and side-chains with the MM-GBSA method. MM-GBSA methods reproduced the experimental relative affinities of the four inhibitors in good agreement (R(2) = 0.996) between experimental and computed binding energies. The MM-GBSA calculations showed that, the number of hydrogen bonds formed between the HDLP and inhibitors, which varied in the system studied, and electrostatic interactions determined the magnitude of the free energies for HDLP-inhibitor interactions. The MM-GBSA calculations revealed that the binding of HDLP to these four hydroxamic acid inhibitors is mainly driven by van der Waals/nonpolar interactions. This study can be a guide for the optimization of HDAC inhibitors and future design of new therapeutic agents for the treatment of cancer. PMID- 18398906 TI - Crystal structure of GH13 alpha-glucosidase GSJ from one of the deepest sea bacteria. AB - The crystal structure of the GH13 alpha-glucosidase (GSJ) from deep-sea bacterium Geobacillus sp. strain HTA-462 was determined to a 2.0 A resolution. Comparisons of the GSJ structure with that of other GH13 enzymes with different catalytic activities revealed that the catalytic cleft of GSJ was widely opened when compared with the homologues. The wide opening of the catalytic cleft originated from conformational changes of active site residues and disorder of the regions close to the catalytic center. This structural feature of GSJ would explain the ability of this enzyme to accept a wide variety of nonsugar molecules as acceptors in the transglycosylation reaction. PMID- 18398907 TI - Structure of the partially unliganded met state of 400 kDa hemoglobin: insights into ligand-induced structural changes of giant hemoglobins. AB - Recent crystallographic studies have revealed the structures of some invertebrate extracellular giant hemoglobins of 3,600 kDa or 400 kDa and their common quaternary structure of dodecameric subassembly composed of four kinds of globin subunits (A1, A2, B1, and B2). These results have provided insight into the mechanisms of their unique functional properties of oxygen binding and sulfide binding. All of these structures were solved with oxygenated or CO-liganded forms at low or moderate resolutions. We have determined the crystal structure of 400 kDa Hb from a polychaete Oligobrachia mashikoi at 1.95 A resolution. The electron densities at higher resolution confirm the existence of an isoform of the B1 subunit because of the inconsistency with the model that was built from the formerly known amino acid sequence. The brownish color of the crystals used in this study and the absorption spectrum from the dissolved crystals strongly indicated that the obtained structure was a ferric met state, whereas complete absence of electron density around the distal heme pockets were observed at the A2, B1, and B2 subunits. We concluded that the obtained structure was in unliganded met forms at three of four globin subunits in the 24mer assembly and in oxygenated forms at the remaining A1 subunits. The partially unliganded structure showed remarkable structural changes at the AB loop regions causing quaternary rearrangements of the EF-dimer structure. In contrast, few changes occurred at the interface regions composed of the E and F helices. These results suggest that the ligand-induced structural changes of Oligobrachia Hb are quite different from those of the well-studied mollusk Hb having the same EF-dimer structure. The structural rearrangements make the dodecameric subassembly form a tighter conformation than those of fully oxygenated or CO-liganded dodecamer structure. PMID- 18398908 TI - Analysis of the sequence and structural features of the left-handed beta-helical fold. AB - The left-handed parallel beta-helix (LbetaH) is a structurally repetitive, highly regular, and symmetrical fold formed by coiling of elongated beta-sheets into helical "rungs." This canonical fold has recently received interest as a possible solution to the fibril structure of amyloid and as a building block of self assembled nanotubular structures. In light of this interest, we aimed to understand the structural requirements of the LbetaH fold. We first sought to determine the sequence characteristics of the repeats by analyzing known structures to identify positional preferences of specific residues types. We then used molecular dynamics simulations to demonstrate the stabilizing effect of successive rungs and the hydrophobic core of the LbetaH. We show that a two-rung structure is the minimally stable LbetaH structure. In addition, we defined the structure-based sequence preference of the LbetaH and undertook a genome-wide sequence search to determine the prevalence of this unique protein fold. This profile-based LbetaH search algorithm predicted a large fraction of LbetaH proteins from microbial origins. However, the relative number of predicted LbetaH proteins per specie was approximately equal across the genomes from prokaryotes to eukaryotes. PMID- 18398909 TI - Crystal structure of PAE0151 from Pyrobaculum aerophilum, a PIN-domain (VapC) protein from a toxin-antitoxin operon. PMID- 18398910 TI - Physico-chemical and technological properties of sodium naproxen granules prepared in a high-shear mixer-granulator. AB - In the present work, authors produced tablets of anhydrous sodium naproxen by wet granulation using a high-shear mixer-granulator. Drug hydrated to the tetrahydrated form, as observed by X-ray powder diffractometry. After wet granulation, authors then performed two different drying procedures, obtaining granules of different water content and crystallographic characteristics. The first procedure dried granules in the high-shear mixer-granulator by applying vacuum at room temperature (batch A), while the second employed the same apparatus and time, under vacuum at 40 degrees C (batch B). X-ray powder diffractometry revealed that the sodium naproxen (SN) contained in batch A granules was a mixture of dihydrated and tetrahydrated forms (as demonstrated by the coexistence of peaks typical of both hydrated forms), while that of batch B granules was a mixture of monohydrated and tetrahydrated forms. This means that differences in drying procedures could lead to products of different crystallographic properties. The behavior under compression was evaluated, revealing that batch A offered the best tabletability and compressibility. These results make it possible to conclude that differences in the crystallographic properties and water content of SN are such that different hydration/drying processes can alter the drug crystal form and thus the tabletability of the resulting granules. PMID- 18398911 TI - Test-retest repeatability of assessing environmental and lifestyle factors in Parkinson's disease. AB - Epidemiological studies of environmental risk factors in Parkinson's disease (PD) are dependent on recollection of past exposures based on patients' self-reports. There are limited studies that have assessed the quality of such data. We conducted a prospective study to determine the test-retest repeatability of environmental and lifestyle factors, and medical data in a PD cohort of Asian ethnicity. A total of 150 consecutive PD patients were initially screened, and 100 were recruited and completed an initial interview. Eighty-three patients completed the second interview more than 6 months later. Lifestyle habits (such as smoking and coffee consumption) showed excellent agreement (kappa > 0.90). For the amount and duration of coffee, tea, alcohol, and cigarette smoking exposure, the total agreement in the response for these factors in the repeat interview were noted in 71.4%, 73.3%, 100%, and 90%, respectively (ICC > 0.83). Medical conditions for which the patients were on treatment, such as diabetes, hypertension, and stroke, revealed very high repeatability (kappa = 0.81-0.90). Environmental exposures like well-water consumption and prior farm-dwelling produced a moderately good repeatability (kappa = 0.66-0.77). In conclusion, our study demonstrates that even over long interval period of more than half a year, self-report lifestyle exposure information, personal and environmental exposure data can be collected with moderate-to-high repeatability from PD patients. PMID- 18398912 TI - A case of parkinsonism worsened by losartan: a probable new adverse effect. PMID- 18398913 TI - Apathy following subthalamic stimulation in Parkinson disease: a dopamine responsive symptom. AB - To evaluate the effects of the dopamine D2-D3 agonist ropinirole in patients who developed apathy after complete withdrawal from dopaminergic medication following successful subthalamic nucleus (STN) stimulation for advanced Parkinson disease (PD). We assessed apathy (Apathy Scale, Apathy Inventory), mood (Montgomery Asberg Depression Rating Scale), cognitive functions (Mattis Dementia rating scale, frontal score, executive tests) and motor state (UPDRS-III) in 8 PD patients treated with STN stimulation without dopaminergic treatment and who became apathetic. Assessments were made at baseline and after 6 weeks of ropinirole treatment (7.2 +/- 5.9 mg/d; range 1-18 mg/d). Apathy improved with ropinirole in all but 1 patient (54 +/- 24%; range 0-78%). Mood also improved (75 +/- 31%; range 0-100%), but not in correlation with the change in apathy. Cognitive performance was not modified. Stimulation contacts were located within the STN in all patients except the one who remained apathetic in spite of ropinirole treatment (zona incerta). We suggest that apathy, which was compensated for by an enhancement of D2-D3 receptor stimulation in PD patients with STN stimulation: (1) depends on a dopaminergic deficit in associativo-limbic areas of the brain and (2) can be avoided if a dopaminergic agonist is administered postoperatively. PMID- 18398914 TI - Geographical variation of medicated parkinsonism in Finland during 1995 to 2000. AB - We performed a nation-wide study on geographical variation in the incidence and prevalence of medicated parkinsonism among the Finns aged > or =30 years using Bayesian spatial conditional autoregressive models. Registry of reimbursed medication for parkinsonism and a prescription database of purchase of these drugs were used to identify the study subjects. They were located by the map coordinates of the place of residence and aggregated into regular 100 km(2) grid cells. A total of 7,190 incident and 10,616 prevalent cases were found. The age adjusted annual incidence was 32.6/100,000 (95% HDR 31.8-33.4) during the years 1995 to 2000 and prevalence was 268/100,000 (95% HDR 263-274) in 2000. The male to female ratio was 1.45 (95% HDR 1.39-1.51) in incidence and 1.54 (95% HDR 1.47 1.61) in prevalence. There was strong evidence for geographic variation in incidence and prevalence. A zone with high incidence and prevalence was identified in the eastern and central part of Finland. There was no evidence for difference in incidence and prevalence between urban and rural areas. The marked (more than two-fold) geographic variation can hardly be caused solely by practices of the registration and collection of data on diagnosis or by methodological issues, but rather suggests to geographic variation in protective and predisposing factors of Parkinsonism in Finland. PMID- 18398915 TI - Low myocardial MIBG uptake in multiple system atrophy with incidental Lewy body pathology: an autopsy case report. PMID- 18398916 TI - The frequency of cardiac valvular regurgitation in Parkinson's disease. AB - To investigate the frequency of cardiac valve regurgitation related with low dose dopamine agonists in patients with Parkinson's disease (PD), echocardiograms were analyzed in 527 consecutive PD patients (448 patients treated with dopamine agonists, 79 patients never treated with dopamine agonists as age-matched controls). The frequency of mild or above mild regurgitation of the aortic valve (AR) was significantly higher in the cabergoline group (13.7%, P < 0.05) compared with the controls (2.5%). Odds ratio adjusted by age and sex for AR was significantly higher in the cabergoline group (OR, 6.45; 95% CI, 1.46-28.60; P = 0.01): odds ratio was significantly higher in patients treated with higher daily doses (OR, 14.41; 95% CI, 3.08-67.38; P = 0.0007) and higher cumulative doses (OR, 15.29; 95% CI, 3.19-73.18; P = 0.0006). No statistical difference was identified in the frequency of the tricuspid and mitral regurgitation. None of the other dopamine agonist groups including pergolide gave higher frequency or higher odds ratio compared with the controls. None of our patients showed severe regurgitation or was operated for valvular heart disease. The question as to whether or not longer duration of low dose dopamine agonist treatment would yield the same results needs further studies. PMID- 18398917 TI - The hand that has forgotten its cunning--lessons from musicians' hand dystonia. AB - Focal task-specific dystonia of the musicians' hand (FTSDmh) is an occupational movement disorder that affects instrumental musicians and often derails careers. There has been speculation on the role of intense practice or the specific technical demands of various instruments as triggers for the development of FTSDmh. In this study, we review the clinical features of all published cases (899 patients) and 61 previously unpublished cases of FTSDmh. Our primary goals were to search for patterns in the clinical phenotype, and to discern if specific instrumental technical demands might be related to the development of dystonia. Symptoms of FTSDmh began at a mean age 35.7 years (SD = 10.6), with an overwhelming male predominance (M:F = 4.1:1). The right hand was preferentially affected in keyboard and plucked string players (77%), and the left hand in bowed string players (68%). Flexion movements were the most common dystonic movement in each instrument class, and fingers 3, 4, and 5, either in isolation or combination, were most frequently involved. The clinical implications of these findings and their possible relationship to the pathophysiology of focal task specific dystonia are explored. PMID- 18398920 TI - Paradoxical kinesia. PMID- 18398918 TI - Mechanisms of hepatocyte growth factor-mediated and epidermal growth factor mediated signaling in transdifferentiation of rat hepatocytes to biliary epithelium. AB - Previous studies from our laboratory have demonstrated that hepatocytes can transdifferentiate into biliary epithelium (BE) both in vivo and in vitro; however, the mechanisms are unclear. The current study was designed to investigate the mechanisms of hepatocyte transdifferentiation in vitro. Rat hepatocytes were cultured in roller bottles to obtain hepatocyte organoid cultures, which were stimulated with various growth factors (GFs) including hepatocyte growth factor (HGF), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), stem cell factor (SCF), macrophage-stimulating protein (MSP), fibroblast growth factor a (FGF-a), fibroblast growth factor-b (FGF-b), and fibroblast growth factor-8b (FGF-8b). Only the cultures treated with HGF, EGF, and their combination exhibited formation of hepatocyte-derived biliary epithelium (BE) despite the presence and activation of all the pertinent cognate membrane receptors of the rest of the GFs. Microarray analysis of the organoid cultures identified specific up-regulation of approximately 500 target genes induced by HGF and EGF, including members of the extracellular matrix (ECM) protein family, Wnt/beta-catenin pathway, transforming growth factor beta (TGF-beta)/bone morphogenetic protein (BMP) pathway, and CXC (cysteine-any amino acid-cysteine) chemokines. To investigate the downstream signaling involved in hepatocyte to biliary epithelial cell (BEC) transdifferentiation, we investigated expression and activities of mitogen-activated protein (MAP) kinases [extracellular signal-regulated kinase (ERK)1/2, p38, and c-Jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK)] as well as serine/threonine kinase AKT. The analysis indicated that AKT phosphorylation was particularly increased in cultures treated with HGF, EGF, and their combination. Whereas phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 completely inhibited biliary epithelium formation, AKT inhibitor could only moderately reduce formation of BE in the organoid cultures treated with HGF+EGF. Most of the HGF+EGF target genes were altered by LY294002. CONCLUSION: Taken together, these data indicate that hepatocyte to BE transdifferentiation is regulated by HGF and EGF receptors and that PI3 kinase-mediated signaling independent of AKT is a crucial component of the transdifferentiation process. PMID- 18398921 TI - Probing exciton diffusion in semiconductors using semiconductor-nanorod quantum structures. PMID- 18398922 TI - One- and two-dimensional diffusion of metal atoms in graphene. PMID- 18398923 TI - Growth kinetics of gold nanocrystals: a combined small-angle X-ray scattering and calorimetric study. AB - The growth of gold nanocrystals prepared by the reduction of tetrachloroauric acid by tetrakis(hydroxymethyl)phosphonium chloride, which allows slow reduction, is investigated by small-angle X-ray scattering and isothermal titration calorimetry in combination with transmission electron microscopy. The growth of the nanocrystals does not follow the diffusion-limited Ostwald ripening but instead follows a sigmoidal rate curve. The activation energy obtained from the temperature-dependent growth study is very small. The heat change associated with the growth is determined for the first time as approximately 10 kcal mol(-1) per 1 nm increase in the nanocrystals' diameter. PMID- 18398924 TI - Striped alloy nanowire optical reflectance barcodes prepared from a single plating solution. PMID- 18398925 TI - Dramatic improvements in toughness in poly(lactide-co-glycolide) nanocomposites. AB - Poly(lactide-co-glycolide) (PLG), a biocompatible and biodegradable polymer, is dramatically toughened by adding small amounts of surface modified clay nanoparticles. The elongation during tensile tests increases from 7% for the pure polymer to 210% for the nanocomposite, accompanied with a modest increase in modulus. In contrast, PLG nanocomposites based on fumed silica treated with hexamethyldisilazane show only modest improvements in toughness. Electron microscopy, X-ray scattering, rheometry, and dielectric relaxation spectroscopy are used to investigate the toughening mechanism. Multiple crazing occurs in the clay nanocomposite after yielding. Small angle X-ray scattering studies show significant orientation of the clay nanoparticles along the tensile stress direction during deformation. The clay nanocomposites show a new, slow relaxation mode, most likely due to interfacial adsorbption of PLG chains on the surface of the clay nanoparticles. The dramatic increase in toughness is attributed to physical crosslinks introduced by the clay nanoparticles, a mechanism absent in the PLG/silica nanocomposites. The physical crosslinks increase the brittle fracture strength of the polymer and, consequently, trigger a toughening mechanism via multiple crazing and shear yielding. PMID- 18398926 TI - Preoperative local staging of endometrial cancer: transvaginal sonography vs. magnetic resonance imaging. AB - OBJECTIVES: To compare the accuracy of transvaginal sonography (TVS) and magnetic resonance imaging (MRI) in the preoperative staging of endometrial carcinoma. METHODS: This was a prospective study in which 74 women consecutively diagnosed with endometrial carcinoma were examined using TVS by physicians trained in gynecological sonography and MRI by radiologists with a special interest in gynecology. All patients underwent surgical-pathological staging after removal of the uterus, adnexa and pelvic lymph nodes. Sensitivity, specificity, and positive and negative predictive values were calculated for each imaging modality with regard to detection of neoplastic invasion of the outer half of the myometrium and cervical involvement. RESULTS: TVS and MRI performed equally well in the preoperative staging of endometrial cancer, with no statistically significant differences between the two techniques. The sensitivity, specificity, positive and negative predictive values, and overall diagnostic accuracy for TVS in the evaluation of myometrial infiltration were 84%, 83%, 79%, 88% and 84%, respectively. Respective values for MRI were 84%, 81%, 77%, 87% and 82%. The corresponding statistics for detection of cervical involvement were 93%, 92%, 72%, 98% and 92% for TVS; and 79%, 87%, 58%, 95% and 85% for MRI. CONCLUSIONS: When carried out by expert practitioners, TVS shows good accuracy in the local staging of endometrial carcinoma. Because of its high costs, MRI should be offered only to those in whom TVS produces images of poor quality. PMID- 18398927 TI - Effects of icariin combined with Panax notoginseng saponins on ischemia reperfusion-induced cognitive impairments related with oxidative stress and CA1 of hippocampal neurons in rat. AB - Previous studies suggest that treatment with icariin (ICA) combined with Panax notoginseng saponins (PNS) improved behavior and cholinergic system disorders followed by amyloid beta-peptide(25-35) lateral ventricle injection in rats. The present study investigated whether administration of ICA + PNS had preventive and therapeutic effects on bilateral common carotid arteries (CCA) occlusion-induced cerebral ischemia-reperfusion (IR) injury in rats. Male Sprague-Dawley rats were divided randomly as follows: sham-operated, i.g. vehicle, ICA (5 mg/kg), PNS (40 mg/kg), ICA + PNS (2.5 + 20, 5 + 40 or 10 + 80 mg/kg), and ergoloid mesylate as a positive control (0.45 mg/kg) in model rats. Treatment was performed once a day for 7 days prior to ischemia. The rats were subjected to transient global IR induced by CCA occlusion in combination with intraperitoneal injection of sodium nitroprusside (2.0 mg/kg), then treated with ICA + PNS for another 14 days continuously. ICA + PNS significantly improved the rat passive avoidance task in step-down paradigms, and spatial cognition in the eight-arm radial maze, concomitant with an improvement of blood viscosity. Increased lipid peroxidation in brain after IR injury was observed, MDA being 0.56 +/- 0.10 nmol/mg prot vs 0.48 +/- 0.06 nmol/mg prot in the vehicle control (p < 0.05). Treatment with ICA + PNS 2.5 + 10, 5 + 40, 10 + 80 mg/kg produced a marked reduction in the MDA level to 0.46 +/- 0.06, 0.42 +/- 0.09 and 0.45 +/- 0.08 nmol/mg prot, respectively vs 0.56 +/- 0.10 nmol/mg prot in IR injury only control (p < 0.05, p < 0.01). A decrease in superoxide dismutase activity was observed in the brain of IR rats (the SOD activity being 72.75 +/- 4.62 U/mg prot vs 80.97 +/- 6.06 U/mg prot in control, p < 0.05). ICA + PNS 5 + 40 mg/kg prevented the IR injury mediated fall in superoxide dismutase activity being 78.90 +/- 6.61 U/mg prot versus 72.75 +/- 4.62 U/mg prot (p < 0.05). ICA + PNS tended to attenuate apoptosis in hippocampal CA1 pyramidal neurons. Either ICA or PNS treatment alone did not obviously improve cognitive impairment (except that lipid peroxidation was reduced by PNS-treatment). The results indicated that ICA + PNS may ameliorate learning and memory deficit and blood viscosity by protecting neurons from oxidative stress in ischemic brain. PMID- 18398928 TI - Antioxidant potential of selected Spirulina platensis preparations. AB - Recent studies suggest that Spirulina, a unicellular blue-green alga, may have a variety of health benefits and therapeutic properties and is also capable of acting as an antioxidant and antiinflammatory agent. In this study, a cell-free and a cell-based test assay were used to examine the antioxidant and antiinflammatory properties of four selected Spirulina platensis preparations: (1) Biospirulina, (2) SpiruComplex, a preparation with naturally bound selenium, chromium and zinc, (3) SpiruZink, a preparation with naturally bound zinc, (4) Zinkspirulina + Acerola, a preparation with naturally bound zinc and acerola powder. The cell-free test assay used potassium superoxide as a donor for superoxide radicals, whereas the cell-based test assay used the formation of intracellular superoxide radicals of functional neutrophils upon stimulation by phorbol-12-myristate-13-acetate as a model to investigate the potential of Spirulina preparations to inactivate superoxide radicals. In accordance with the recommended daily dosage, test concentrations ranging from 50 to 1000 microg/mL were chosen. The results showed a dose-dependent inactivation of free superoxide radicals (antioxidant effect) as well as an antiinflammatory effect characterized by a dose-dependent reduction of the metabolic activity of functional neutrophils and a dose-dependent inactivation of superoxide radicals generated during an oxidative burst. The results demonstrate that the tested Spirulina preparations have a high antioxidant and antiinflammatory potential. Especially SpiruZink and Zinkspirulina + Acerola might be useful as a supportive therapeutic approach for reducing oxidative stress and/or the generation of oxygen radicals in the course of inflammatory processes. PMID- 18398929 TI - Immunomodulatory effect of extracts of Hibiscus sabdariffa L. (Family Malvaceae) in a mouse model. AB - The immunomodulatory activity of water and alcohol extracts (including its fractions) of the dried calyx of the plant was evaluated in mice. The ability of the extracts to inhibit or enhance the production of two cytokines, namely tumor necrosis factor-alpha (TNF- alpha) and interleukin-10 (IL-10), respectively, implicated as proinflammatory and antiinflammatory interleukins were also evaluated. The extracts at doses of 50 mg/kg were found to possess higher immunostimulatory activities in comparison with levamisole (positive control), with significant effects when compared with the vehicle-treated group (p < 0.01). Increased activity was observed with increase in doses of the 50% ethanol and absolute ethanol extracts. The insoluble fraction exhibited a significant dose dependent immunostimulatory activity (p < 0.05), while the residual water-soluble fraction exhibited activity at 100 mg/kg body weight. The production of tumor necrosis factor-alpha (TNF-alpha), was low in all the extract groups tested, while the production of interleukin 10 (IL-10) was high compared with the control. The production of IL-10 was high in 300 mg/kg aqueous extract. The insoluble fraction exhibited a profound dose-dependent immunostimulatory activity higher than the positive control at 100 mg/kg. This study established the immunoenhancing properties of the extracts of this plant confirming that the immunomodulatory activity is cell mediated and humoral. The insoluble fraction could find use as an immunostimulatory agent in humans. PMID- 18398930 TI - Effective clearance of intracellular Leishmania major in vivo requires Pten in macrophages. AB - Leishmaniases are a major international public health problem, and macrophages are crucial for host resistance to this parasite. To determine if phosphatase and tensin homologue deleted on chromosome ten (Pten), a negative regulator of the PI3K pathway, plays a role in macrophage-mediated resistance to Leishmania, we generated C57BL/6 mice lacking Pten specifically in macrophages (LysMCrePten(flox/flox) mice). Examination of lesions resulting from Leishmania major infection showed that LysMCrePten(flox/flox) mice were more susceptible to the parasite than wild-type (WT) mice in the early phase of the infection, but were eventually able to eliminate the pathogen. In vitro Pten-deficient macrophages showed a reduced ability to kill parasites in response to IFN-gamma treatment, possibly because the mutant cells exhibited decreased TNF secretion that correlated with reductions in inducible nitric oxide synthase expression and nitric oxide production. In response to various TLR ligands, Pten-deficient macrophages produced less TNF and IL-12 but more IL-10 than WT cells. However, analysis of cells in the lymph nodes draining L. major inoculation sites indicated that both LysMCrePten(flox/flox) and WT mice developed normal Th1 responses following L. major infection, in line with the ability of LysMCrePten(flox/flox) mice to eventually eliminate the parasite. Our results indicate that the efficient clearance of intracellular parasites requires Pten in macrophages. PMID- 18398931 TI - Simultaneous presentation and cross-presentation of immune-stimulating complex associated cognate antigen by antigen-specific B cells. AB - We demonstrate that uptake of oligomeric cognate antigen (OVA-hen egg lysozyme, OVA-HEL) alone or incorporated in immune-stimulating complexes (ISCOMS) facilitates presentation and simultaneous cross-presentation of OVA by HEL specific B cells in vitro. HEL-specific B cells stimulated CD8(+) T cell responses in vitro to the same extent as bone marrow-derived dendritic cells. Cross-presentation by specific B cells required endosomal acidification, proteasomal processing and classical MHC class I/peptide transport. Specific B cells also acquired both antigens rapidly in vivo and presented them to CD4(+) T cells. However, only HEL-specific B cells from OVA-HEL ISCOMS-immunised mice could cross-present OVA to naive OVA-specific CD8(+) T cells. Antigen-specific B cells were also activated selectively by OVA-HEL ISCOMS in vitro and importantly, the presence of HEL-specific B cells promoted the persistence of clonal expansion of OVA-specific CD8(+) T cells after in vivo immunisation with OVA-HEL ISCOMS. These results demonstrate preferential MHC class I and class II processing of cognate antigen incorporated in ISCOMS by specific B cells in vitro and in vivo, highlighting the ability of ISCOMS to target B cells and offering novel insights into the role of B cells in cross-presentation to CD8(+) T cells. PMID- 18398932 TI - Monocyte-dependent oncostatin M and TNF-alpha synergize to stimulate unopposed matrix metalloproteinase-1/3 secretion from human lung fibroblasts in tuberculosis. AB - Leukocyte-derived matrix metalloproteinases (MMP) are implicated in the tissue destruction characteristic of tuberculosis (TB). The contribution of lung stromal cells to MMP activity in TB is unknown. Oncostatin M (OSM) is an important stimulus to extrapulmonary stromal MMP induction, but its role in regulation of pulmonary MMP secretion or pathophysiology of TB is unknown. We investigated OSM secretion from Mycobacterium tuberculosis (Mtb)-infected human monocytes/macrophages and the networking effects of such OSM on lung fibroblast MMP secretion. Mtb increased monocyte OSM secretion dose dependently in vitro. In vivo tuberculous granulomas immunostained positively for OSM. Further, conditioned media from Mtb-infected monocytes (CoMTb) induced monocyte OSM secretion (670 +/- 55 versus 166 +/- 14 pg/mL in controls), implicating an autocrine loop. Mtb-induced OSM secretion was prostaglandin (PG) sensitive, and required activation of surface G-protein coupled receptors. OSM induction was ERK MAP kinase dependent, p38-requiring but JNK-independent. OSM synergized with TNF alpha, a key cytokine in TB granuloma formation, to stimulate pulmonary fibroblast MMP-1/-3 secretion, while suppressing secretion of tissue inhibitors of metalloproteinases-1/-2. In summary, Mtb infection of monocytes results in PG dependent OSM secretion, which synergizes with TNF-alpha to drive functionally unopposed fibroblast MMP-1/-3 secretion, demonstrating a previously unrecognized role for OSM in TB. PMID- 18398933 TI - Antigen structure influences helper T-cell epitope dominance in the human immune response to HIV envelope glycoprotein gp120. AB - The development of an effective vaccine against HIV/AIDS has been hampered, in part, by a poor understanding of the rules governing helper T-cell epitope immunodominance. Studies in mice have shown that antigen structure modulates epitope immunodominance by affecting the processing and subsequent presentation of helper T-cell epitopes. Previous epitope mapping studies showed that the immunodominant helper T-cell epitopes in mice immunized with gp120 were found flanking flexible loops of the protein. In this report, we show that helper T cell epitopes against gp120 in humans infected with HIV are also found flanking flexible loops. Immunodominant epitopes were found to be located primarily in the outer domain, an average of 12 residues C-terminal to flexible loops. In the less immunogenic inner domain, epitopes were found an average of five residues N terminal to conserved regions of the protein, once again placing the epitopes C terminal to flexible loops. These results show that antigen structure plays a significant role in the shaping of the helper T-cell response against HIV gp120 in humans. This relationship between antigen structure and helper T-cell epitope immunodominance may prove to be useful in the development of rationally designed vaccines against pathogens such as HIV. PMID- 18398934 TI - Mast cell-mediated immune responses through IgE antibody and Toll-like receptor 4 by malarial peroxiredoxin. AB - In this study, 2-Cys Plasmodium berghei ANKA (PbA) peroxiredoxin (Prx) was identified as an antigenic protein recognized by an anti-PbA IgE antibody using two-dimensional polyacrylamide gel electrophoresis and proteomic analysis. Innate immune responses to PbAPrx were examined using cells from mice deficient in Toll like receptors (TLR) or related molecules, and it was demonstrated that responses were severely impaired in TLR4(-/-), MyD88(-/-) and MD-2(-/-) mice, but not in Toll/IL-1 receptor domain-containing adaptor inducing IFN-gamma (TRIF)(-/-), TLR2(-/-) or radioprotective 105 (RP105)(-/-) mice. An association between PbAPrx and TLR4 was observed following immunoprecipitation and immunoblotting, suggesting that PbAPrx was associated with TLR4/MD-2. Interactions between Prx and TLR4/MD-2 were also examined by flow cytometry using TLR4/MD-2- or TLR2 expressing cells. NFkappaB/GFP activity was observed in TLR4/MD-2- but not in TLR2-expressing cells following stimulation with Prx. However, this effect was not observed after treatment with proteinase K, suggesting that PbAPrx is a protein ligand for TLR4 and that the PbAPrx activity observed in this study is not due to contamination with LPS. These findings indicate that malarial Prx induces IgE-mediated protection through FcepsilonRI on mast cells and innate immunity through TLR4 with MyD88 and MD-2, suggesting a novel function for malarial Prx in innate and acquired immune responses in malaria. PMID- 18398935 TI - The use of central nervous system drugs and analgesics among very old people with and without dementia. AB - PURPOSE: Old people in general, and particularly those with dementia, are more sensitive to adverse drug effects than younger people. Despite this, the use of central nervous system (CNS) drugs among old people is common. The aim of the present study was to compare the use of central nervous system drugs and analgesics among people aged 85 years or older, with and without dementia. METHODS: One half of all people aged 85 years old and all those aged 90, 95 and above in Umea, Sweden and Vaasa and Mustasaari, Finland, were asked to participate in this part of the GERDA/Umea 85+ study. Both those living in their own homes and those in institutions were included. Trained investigators performed structured interviews and assessments. Medication data were obtained from the participants and medical records. Dementia disorders were diagnosed according to DSM-IV. RESULTS: Dementia was diagnosed in 247/546 participants (45.2%). A higher proportion of the participants with dementia used paracetamol (50.6% compared to 21.4%, p < 0.001), antipsychotics (22.3% and 2.7%, p < 0.001), antidepressants (33.6% and 11.4%, p < 0.001) and anxiolytics (19.0% and 8.0%, p < 0.001). There were no differences in the use of opioid analgesics, anticonvulsants and hypnotics. CONCLUSIONS: The use of CNS drugs and analgesics was common among this population of very old people. Furthermore, the prescription of CNS drugs was more common among people with dementia. The use of antipsychotics in people with dementia should arouse particular concern, because of the high risk of severe adverse events and the limited evidence of positive effects. PMID- 18398937 TI - Global damage in systemic juvenile idiopathic arthritis: preliminary early predictors. AB - OBJECTIVE: To assess damage in systemic juvenile idiopathic arthritis (sJIA) by the use of the Juvenile Arthritis Damage Index (JADI) and to identify early predictors of global, articular, and extraarticular damage. METHODS: Forty-seven consecutive patients with sJIA with a disease duration > 24 months were assessed for damage in a cross-sectional evaluation. The JADI was administered by 2 pediatric rheumatologists. Damage was defined as JADI score >or= 1. Early clinical variables were retrieved from clinical records, and they included demographic, clinical, and laboratory characteristics. Univariate analysis was used to select candidate predictors to be included in multiple logistic regression. RESULTS: Twenty (43%) patients exhibited damage: 18 (38%) patients had articular and 9 (19%) extraarticular damage. JADI score ranged between 0 and 24. Cervical spine arthritis and corticosteroid usage occurring in the first 6 months of the disease course were found as predictors of damage. Damage scores correlated with number of joints with limited motion, and with functional disability. CONCLUSION: Articular damage is the main component of global damage in patients with sJIA. Early cervical spine involvement and corticosteroid usage may identify patients with sJIA at risk of developing damage. PMID- 18398936 TI - Pregnancy outcome in juvenile systemic lupus erythematosus: a Brazilian multicenter cohort study. AB - OBJECTIVE: To determine pregnancy outcome and fetal loss risk factors in patients with juvenile systemic lupus erythematosus (JSLE). METHODS: A total of 315 female patients with JSLE followed in 12 Brazilian pediatric rheumatology centers were consecutively selected. Menarche was observed in 298 (94.6%) patients. Patients' medical records were reviewed for pregnancy outcomes and demographic, clinical, and therapeutic data. RESULTS: A total of 24 unplanned pregnancies occurred in 298 (8%) patients. The outcomes were 5 (21%) early fetal losses (prior to 16 wks gestation), 18 (75%) live births, and 1 (4%) death due to preeclampsia and premature birth. The frequencies of active diffuse proliferative glomerulonephritis, proteinuria > or = 0.5 g/day, and arterial hypertension at the beginning of pregnancy were higher in pregnancies resulting in fetal losses than in live births [60% vs 5% (p = 0.02), 60% vs 5% (p = 0.02), 60% vs 5% (p = 0.02), respectively]. JSLE pregnancies with fetal losses had a significantly higher mean SLE Disease Activity Index 2000 (SLEDAI-2K) at the start of pregnancy compared with those with live births (9.40 +/- 7.47 vs 3.94 +/- 6.00; p = 0.049). Four pregnancies were inadvertently exposed to intravenous cyclophosphamide therapy for renal involvement despite contraceptive prescriptions, resulting in fetal loss in 3 (p = 0.02). In multivariate analysis only intravenous cyclophosphamide use at start of pregnancy (OR 25.50, 95% CI 1.72-377.93, p = 0.019) remained as an independent risk factor for fetal loss. CONCLUSION: We identified immunosuppressive therapy as the major contributing factor for fetal loss in JSLE, reinforcing the importance of contraception. PMID- 18398938 TI - Intraarticular corticosteroid injections of the temporomandibular joint in juvenile idiopathic arthritis. AB - OBJECTIVE: To describe the clinical and radiographic outcomes in a series of patients with juvenile idiopathic arthritis (JIA) who underwent one or more intraarticular corticosteroid (IAS) injections of the temporomandibular joint (TMJ) performed without imaging guidance. METHODS: Retrospective chart review was performed for all patients with JIA diagnosed and treated at our institution between January 1, 2000, and January 1, 2006, who underwent one or more IAS injections of their TMJ. IAS injections were performed by the same oral and maxillofacial surgeon without imaging guidance, using either triamcinolone acetonide or triamcinolone hexacetonide. The primary outcomes assessed were maximal incisal opening (MIO) measurements, patient-reported symptoms, physical examination findings, and imaging results. RESULTS: Twenty-five patients were identified. Twenty-one (84%) had radiographic evidence of TMJ disease when TMJ disease was first suspected by their physician. The 25 patients underwent 74 IAS injections on 47 separate occasions. When baseline MIO measurements were compared to the last MIO measurements of the study period, there was a mean increase in MIO of 6.9 mm (p = 0.002; 95% CI 3, 10.7). There was a mean increase in MIO of 3.8 mm following each IAS injection (p = 0.003; 95% CI 1.4, 6.2). Patients who underwent multiple IAS injections had a mean increase in MIO after first injection of 6.6 mm (p < 0.001; 95% CI 4.1, 9.1); however, the mean increase in MIO after subsequent injections was 0.4 mm (p = 0.8; 95% CI -3.5, 4.4). One patient developed subcutaneous atrophy at the injection site. Two patients developed small, asymptomatic intraarticular calcifications. No additional adverse events were reported. CONCLUSIONS: In this patient population, there was an overall increase in MIO measurements following initial IAS injection and during the study period. Patients tended to have minimal response to subsequent injections. IAS injections performed without imaging guidance by an experienced oral and maxillofacial surgeon were well tolerated with only rare adverse events. The presence of radiographic changes when the physician first suspected TMJ disease in 84% of patients emphasizes the need for better screening and early intervention for synovitis in this joint. PMID- 18398939 TI - Abnormal transforming growth factor-beta expression in mesenchymal stem cells from patients with osteoarthritis. AB - OBJECTIVE: To determine the expression of the genes that code for the isoforms of transforming growth factor-beta (TGF-beta) and TGF-beta receptors (TBR) in mesenchymal stem cells (MSC) from patients with osteoarthritis (OA). METHODS: Total RNA was extracted from primary cultures of MSC and quantitative real-time reverse transcription-polymerase chain reaction was performed to analyze gene expression. RESULTS: MSC from patients with OA showed significantly increased total TGF-beta, TGF-beta1 isoform, TBR-II, and TBR-III mRNA expression compared to controls. CONCLUSION: Our study is the first reporting the gene expression levels of TGF-beta and its isoforms and receptors in patients with OA. These findings might have pathological significance for OA disease. PMID- 18398940 TI - Validation of self-report of rheumatoid arthritis and systemic lupus erythematosus: The Women's Health Initiative. AB - OBJECTIVE: The Women's Health Initiative (WHI), initiated in 1993, enrolled 161,808 postmenopausal women aged 50-79 years and followed them with annual questionnaires for 8 years in order to study major causes of morbidity and mortality. Our objective was to determine the most effective and efficient means to validate self-reported rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) in the WHI. METHODS: Data from 2 of 40 WHI clinical centers were used. Of these 7443 women, 643 self-reported RA and 106 self-reported SLE. Research coordinators contacted these women using mailers and telephone calls to obtain medical record releases and a Connective Tissue Screening Questionnaire (CSQ). Medical records were obtained on 286 self-reported RA and 34 self-reported SLE and reviewed by 3 rheumatologists blind to the self-reported diagnoses. Sensitivity, specificity, and the kappa statistic were computed to evaluate the level of agreement between self-report and chart review. RESULTS: Self-reported RA was accurate only 14.7% (42/286 cases) of the time. Coupling the self-report to medication data improved the positive predictive value (PPV; 62.2%) and kappa (0.53), suggesting a moderate agreement to chart review. Self-reported SLE was accurate only 11.8% (4/34 cases) of the time. Coupling the self-report to medication data improved the PPV (40.0%) and kappa (0.44), suggesting a moderate agreement to chart review. The CSQ was inferior to using medication data but was substantially better than self-report alone. CONCLUSION: The performance of disease self-report coupled with medication history in validating RA and SLE was very good and should obviate the need for time-consuming medical record reviews. PMID- 18398941 TI - Abnormal histone modification patterns in lupus CD4+ T cells. AB - OBJECTIVE: To investigate alterations in histone modifications in patients with systemic lupus erythematosus (SLE). METHODS: Global histone H3/H4 acetylation and H3K4/H3K9 methylation in CD4+ T cells from 20 SLE patients and 10 healthy control subjects were assayed using the EpiQuik global histone H3/H4 acetylation and H3K4/H3K9 methylation assay kits. mRNA levels of 12 members of 3 classes of chromatin modifier genes were measured by real-time quantitative polymerase chain reaction. RESULTS: Global histone H3 and H4 hypoacetylation was observed in active lupus CD4+ T cells compared with controls (p = 0.002 and p = 0.009, respectively). The degree of histone H3 acetylation correlated negatively with increased disease activity in lupus patients as measured by SLEDAI (r = -0.889, p = 0.044). We found global histone H3K9 hypomethylation in both active and inactive lupus CD4+ T cells, compared with controls (p = 0.001, p = 0.003, respectively). However, global levels of H3K4 methylation were not different between patients and controls. SIRT1 mRNA levels were significantly increased in active lupus CD4+ T cells compared with controls (p < 0.001), while mRNA levels of CREBBP, P300, HDAC2, HDAC7, SUV39H2, and EZH2 were significantly downregulated in patients with active lupus (p < 0.001, p < 0.001, p = 0.01, p < 0.001, p = 0.003, p = 0.001, respectively). CONCLUSION: Histone modifications appear abnormal in CD4+ T cells in SLE. PMID- 18398942 TI - Climatic influence on the prevalence of noncutaneous disease flare in systemic lupus erythematosus in Hong Kong. AB - OBJECTIVE: It is generally agreed that there is a seasonal variation in the prevalence of cutaneous manifestations of systemic lupus erythematosus (SLE). We investigated whether there is seasonal variation in the incidence of noncutaneous lupus flare in Hong Kong. METHODS: We reviewed all noncutaneous lupus flare in 222 consecutive patients with SLE followed in our clinic from 1995 to 2005. Specific organ involvement of each flare was reviewed. The variation in the prevalence of lupus flare by calendar month and the relation with climatic factors were determined. RESULTS: The total followup was 18,412 patient-months. In total, there were 313 episodes of noncutaneous flare recorded in 129 patients. There were more lupus flares in December and January [2.31 episodes, vs 1.58 episodes per 100 patient-months for other calendar months; relative risk (RR) 1.46, 95% CI 1.12-1.90, p = 0.004], and more flares of lupus nephritis in December and January (1.14 episodes, vs 0.60 episodes per 100 patient-months for other calendar months; RR 1.90, 95% CI 1.29-2.80, p = 0.001). There were more cases of membranous nephropathy in December and January (0.46 episode, vs 0.18 episode per 100 patient-months for other calendar months; RR 2.59, 95% CI 1.36 4.93, p = 0.0027), while the variation in prevalence of proliferative lupus nephritis was not statistically significant. There was also a significant U-shape correlation between the rate of lupus flare and the monthly average environmental temperature (r = 0.802, p = 0.0096), with higher flare rate at extremes of temperature. CONCLUSION: We found substantial seasonal variation in the incidence of noncutaneous flare in our SLE patients, with peak incidence in December and January. There was a U-shaped relation between environmental temperature and the prevalence of noncutaneous flare. Keeping a warm living environment and avoiding exposure to extremes of temperature may help to reduce flare for SLE patients in subtropical countries. PMID- 18398943 TI - Longterm clinical and immunological effects of anti-CD20 treatment in patients with refractory systemic lupus erythematosus. AB - OBJECTIVE: To retrospectively evaluate longterm clinical and immunological effects of anti-CD20 treatment in patients with systemic lupus erythematosus (SLE) with active nephritis or autoantibody-mediated cytopenias refractory to conventional immunosuppressive treatment. METHODS: Anti-CD20 treatment (rituximab) was added to the ongoing immunosuppressive treatment in 31 SLE patients with active nephritis (n = 17), thrombocytopenia (n = 10), and hemolytic anemia (n = 4) refractory to conventional therapy. Disease activity was evaluated by the SLE Disease Activity Index. The median followup time after anti-CD20 treatment was 22 months (range 1-61 mo). RESULTS: Complete B cell depletion was obtained in all patients. In 11 of the 17 lupus nephritis patients complete or partial responses were achieved after 6-12 months. Eight of these patients increased their glomerular filtration rate (GFR) by > 25%. The responders were characterized by having shorter nephritis duration, a baseline GFR > 30 ml/min, and detectable circulating CD19+ B lymphocytes before B cell depletion. Anti-CD20 treatment was highly effective in patients with autoimmune thrombocytopenia, inducing a significant increase of platelet counts after 1 month (p < 0.01). Five of 10 patients achieved complete normalization of their platelet counts within 6 months. The anti-CD20 treatment was followed by a significant reduction of autoantibodies against dsDNA and platelets, in nephritic and in thrombocytopenic patients, respectively. CONCLUSION: Addition of anti-CD20 treatment to conventional immunosuppressive therapy may be a beneficial strategy in refractory lupus nephritis and autoimmune cytopenias, possibly by reducing the levels of pathogenic autoantibodies. PMID- 18398944 TI - Comparison of an interferon-gamma assay with tuberculin skin testing for detection of tuberculosis (TB) infection in patients with rheumatoid arthritis in a TB-endemic population. AB - OBJECTIVE: Tuberculosis (TB) in patients with rheumatoid arthritis (RA) undergoing treatment with anti-tumor necrosis factor (TNF) agents is commonly the result of reactivation of latent TB infection (LTBI); detection and treatment of LTBI is essential before treatment with anti-TNF agents. We reported previously that the tuberculin skin test (TST) is inaccurate for diagnosis of LTBI in patients with RA. Here, we compare the prevalence of LTBI in RA patients and matched controls according to positive TST and QuantiFeron-TB Gold In-Tube version (QFT) results and determine their agreement. METHODS: A cross-sectional study of 101 RA patients and 93 controls was conducted in Lima, Peru, where the prevalence of LTBI in the general population has been estimated to be 68%. Blood was drawn for QFT assay followed by TST using 2-TU of RT 23 purified protein derivative. TST was deemed positive at >or= 5 mm for RA patients and >or= 10 mm for controls. RESULTS: There were no significant differences between RA patients and controls for age, sex, bacillus Calmette-Guerin vaccination, or history of or contact with TB. 88% of patients had active RA disease and 2 (1.9%) patients had indeterminate QFT results. The number of subjects testing positive with the QuantiFeron assay was comparable between patients and controls (44.6% vs 59.1%, respectively), whereas the TST detected significantly less LTBI among RA patients (26.7%) than controls (65.6%). Thus, the rate of LTBI in RA patients represented 75% and 41% of the rate in their controls using QFT or TST, respectively (p = 0.008). Poor agreement between TST and QFT was seen in RA patients, but in controls, good agreement was observed between these tests. CONCLUSION: In a TB endemic population, the QuantiFeron-TB Gold In-Tube assay seemed to be a more accurate test for detection of LTBI in RA patients compared with the TST, and may potentially improve the targeting of prophylactic therapy before treatment with anti-TNF agents. PMID- 18398945 TI - Prevalence and significance of anti-peptidylarginine deiminase 4 antibodies in rheumatoid arthritis. AB - OBJECTIVE: To detect anti-peptidylarginine deiminase 4 (PAD4) antibody in patients with rheumatoid arthritis (RA) and to investigate its clinical significance in the pathogenesis of RA. METHODS: Serum samples were obtained from 109 patients with RA, 67 systemic lupus erythematosus (SLE), 48 primary Sjogren's syndrome (pSS), 41 systemic sclerosis (SSc), 34 osteoarthritis (OA), 23 dermatomyositis/polymyositis (DM/PM), and 19 ankylosing spondylitis (AS) and 106 healthy individuals. The presence of antibodies against recombinant human PAD4 (anti-PAD4) was examined by ELISA. Associations between anti-PAD4 and the clinical features of RA were evaluated. RESULTS: The prevalence of anti-PAD4 in RA patients (45.0%) was significantly higher than those of SLE (9.0%), pSS (4.2%), SSc (9.8%), OA (5.9%), DM/PM (13.0%), AS (0%), and controls (4.7%). The mean titer of anti-PAD4 in RA was also significantly higher than in SLE, other rheumatic diseases, and controls. Disease Activity Score-28 (DAS28), anti-cyclic citrullinated peptide (CCP) antibody, erythrocyte sedimentation rate, rheumatoid factor, IgM, and IgG in anti-PAD4-positive patients were all higher than in anti PAD4-negative patients. There were positive correlations between anti-PAD4 and DAS28 score (r = 0.333, p < 0.01) and anti-CCP antibody (r = 0.248, p < 0.05). CONCLUSION: The presence of anti-PAD4 in RA indicates that PAD4 may act as an autoantigen that may play a role in the pathogenesis of RA. PMID- 18398946 TI - Antibodies against mutated citrullinated vimentin are a better predictor of disease activity at 24 months in early rheumatoid arthritis than antibodies against cyclic citrullinated peptides. AB - OBJECTIVE: To evaluate the predictive values for disease progression of various antibodies against citrullinated peptide proteins (ACPA) and their relation to PTPN22 1858C/T polymorphism and HLA-DRB1 alleles in early rheumatoid arthritis (RA). METHODS: The ACPA, e.g., antibodies against mutated citrullinated vimentin (MCV), cyclic citrullinated peptides (CCP) type 2 and 3 (both of IgG isotype) and 3.1 (of both IgG and IgA isotypes), were analyzed at baseline in patients with early RA (n = 210) and in population controls (n = 102) using an enzyme immunoassay. A receiver-operating characteristic curve was constructed for each antibody. Disease activity [swollen and tender joints, visual analog scale for global health, and erythrocyte sedimentation rate (ESR)] was evaluated at baseline and regularly for 24 months. Radiographs of hands and feet were graded using the Larsen score. RESULTS: Patients with anti-MCV antibodies had significantly less reduction in Disease Activity Score (DAS28) over time (p < 0.01), and significantly increased area under the curve (AUC) for DAS28 (p < 0.05), ESR (p < 0.01), C-reactive protein (p < 0.01), and swollen joint count (p = 0.057) compared to those without. Corresponding differences were not found in patients with anti-CCP2, CCP3, and CCP3.1 antibodies. Radiological progression (p < 0.0001-0.01) and radiological outcome (p < 0.0001-0.01) at 24 months were significantly predicted by all ACPA after baseline adjustments. PTPN22 T variant and HLA-DRB1 alleles were not related to radiological progression or inflammatory activity over time. CONCLUSION: Anti-MCV antibodies are associated with a more severe RA disease, as measured by DAS28, ESR, and swollen joint count over time, compared with anti-CCP2, CCP3, and CCP3.1 antibodies. Radiological progression was predicted equally by all 4 autoantibodies. PMID- 18398947 TI - Osteonecrosis of the jaw: new developments in an old disease. PMID- 18398948 TI - Pharmacogenomics of methotrexate in rheumatoid arthritis: does race make a difference? PMID- 18398949 TI - The time has come to include assessment of radiographic progression in juvenile idiopathic arthritis clinical trials. PMID- 18398950 TI - Tophaceous calcium pyrophosphate dihydrate deposition disease of the temporomandibular joint. AB - Tophaceous pseudogout is a rare manifestation of calcium pyrophosphate dihydrate (CPPD) deposition disease that particularly affects the temporomandibular joint (TMJ). We describe a case of tophaceous pseudogout and review the literature. Thirty-four cases of chronic CPPD deposition disease affecting the TMJ are described. Symptoms usually included pain and swelling. Most patients required surgery because of extensive crystal deposits, usually localized to the joint and adjacent structures but occasionally invasive. For many patients, malignancy was the preoperative diagnosis. Although patients with acute pseudogout of the TMJ may have involvement of other joints, tophaceous pseudogout was predominantly isolated to the TMJ. PMID- 18398951 TI - Severe back pain in a 56-year-old woman. PMID- 18398953 TI - Progressive multifocal leukoencephalopathy in a minimally immunosuppressed patient with systemic lupus erythematosus treated with dapsone. PMID- 18398952 TI - Prospective study of the association between NAT2 gene haplotypes and severe adverse events with sulfasalazine therapy in patients with rheumatoid arthritis. PMID- 18398954 TI - Pseudotumoral presentation of calcium pyrophosphate dihydrate crystal deposition disease. PMID- 18398955 TI - Activation function-1 domain of estrogen receptor regulates the agonistic and antagonistic actions of tamoxifen. PMID- 18398956 TI - Q &A. Andrew D. Ellington. PMID- 18398957 TI - Neuroblastoma is a prototype of those malignancies which can progress very rapidly but can also be differentiated into a benign variant. In this issue. PMID- 18398958 TI - Re: AAPL practice guideline for the forensic psychiatric evaluation of competence to stand trial. PMID- 18398959 TI - [Imaging in lymphomatoid granulomatosis]. PMID- 18398960 TI - Proceedings of the 12th International Congress on Hormonal Steroids and Hormones & Cancer. September 13-16, 2006. Athens, Greece. PMID- 18398961 TI - WHO: 60 years on. PMID- 18398962 TI - WHO AFRO: failing the region. PMID- 18398963 TI - "I was the first": revisiting a paediatric heart transplantation. PMID- 18398964 TI - Fracture repair: challenges and opportunities. Proceedings of an American Academy of Orthopaedic Surgeons symposium. April 25-28, 2007. PMID- 18398965 TI - Additional abstracts from the XII International Workshop on Chronic Lymphocytic Leukemia. September 14-16, 2007. London, United Kingdom. PMID- 18398966 TI - [Abstracts of the 89th Congress of the Association of Morphologists, 7 June 2007, Limoges, France]. PMID- 18398967 TI - Following Z359 is the best approach. PMID- 18398968 TI - Political theory. PMID- 18398969 TI - Description of a "trans-Saharan" strain of human T-lymphotropic virus type 1 in West Africa. AB - The aim of this study was to assess the prevalence and the molecular epidemiology of human T-lymphotropic virus type 1 (HTLV-1) in a group of pregnant women living in Guinea Bissau. We studied 427 consecutive pregnant women attending 10 centers for HIV-1 infection monitoring in Bissau. HTLV-1 infection was found in 2.6% of the patients. Phylogenetic analysis of the long terminal repeat region showed that 10 isolates were of the cosmopolitan subtype (HTLV-1a) and that only 1 was of the widespread Central African subtype (HTLV-1b). All the cosmopolitan isolates belonged to the HTLV-1aD subgroup, which was first described in North Africa and clustered with other Senegal and Guinea isolates to form a significant West African clade. Our data show a high prevalence of HTLV-1 in Guinea Bissau and suggest the existence of a trans-Saharan strain distributed in North and West Africa, which probably crossed the desert in the past as a result of contacts between nomadic and sedentary populations or along trading routes. PMID- 18398970 TI - Clinical and genetic determinants of intracellular tenofovir diphosphate concentrations in HIV-infected patients. AB - BACKGROUND: Nucleos(t)ide reverse transcriptase inhibitors (NRTIs), such as tenofovir, require intracellular phosphorylation for pharmacologic activity. Drug transporters may contribute to the intracellular disposition of NRTIs. OBJECTIVE: We characterized intracellular tenofovir diphosphate (TFV-DP) concentrations in HIV-infected patients (n = 30), and investigated associations between TFV-DP concentrations and polymorphisms in the drug transporter genes SLC22A6, ABCC2, and ABCC4. METHODS: Subjects were genotyped for 6 single-nucleotide polymorphisms: 2 in SLC22A6 (encodes influx transporter, human organic anion transporter 1), 728G>A and 453G>A; 2 in ABCC2 (encodes efflux transporter, multidrug resistance protein [MRP] 2), -24C>T and 1249G>A; and 2 in ABCC4 (encodes efflux transporter, MRP4), 3463A>G and 4131T>G. RESULTS: The mean TFV-DP was 76.1 fmol/10(6) cells (range: 16.3 to 212 fmol/10(6) cells). Tenofovir apparent oral and renal clearances were significantly predictive of intracellular TFV-DP concentrations. For every 1-L/h decrease in tenofovir renal clearance, there was, on average, an 8% increase in TFV-DP (P = 0.002). We identified a novel relation between ABCC4 3463A>G genotype and TFV-DP. ABCC4 3463G variants had TFV-DP concentrations 35% higher (29 fmol/10(6) cells) than wild type (P = 0.04). CONCLUSION: This study provides direction for future investigations to elucidate the contribution of clinical characteristics and drug transporter genotype to TFV-DP safety and efficacy. PMID- 18398971 TI - Changes in lipid profile over 24 months among adults on first-line highly active antiretroviral therapy in the home-based AIDS care program in rural Uganda. AB - BACKGROUND: Use of highly active antiretroviral therapy (HAART) has been linked to dyslipidemia and increased risk of cardiovascular disease (CVD) in HIV infected patients in industrialized countries. The effects of HAART on lipid metabolism among sub-Saharan Africans, for whom access to antiretroviral therapy is expanding, remain largely unknown. METHODS: From July 2003 to May 2004, 987 antiretroviral-naive patients with symptomatic HIV disease or a CD4 count <250 cells/mm3 were started on HAART in the Home-Based AIDS Care (HBAC) Program in Tororo, Uganda. The HBAC Program provided weekly drug delivery and field-based clinical monitoring. Nonfasting repository sera from a subset of 374 patients were analyzed for levels of total cholesterol (TC), direct low-density lipoprotein cholesterol (LDL-c), direct high-density lipoprotein cholesterol (HDL c), and triglycerides (TG) at baseline (before HAART) and after 12 and 24 months of HAART using Randox enzymatic kits (Crumlin, United Kingdom). RESULTS: The 374 patients evaluated (49% women, mean age = 39 years, CD4 count = 124 cells/mm3, body mass index = 19.7 kg/m2) received initial HAART composed of stavudine, lamivudine, and either nevirapine (365 patients [98%]) or efavirenz (9 patients [2%]). During 24 months, 99 (26%) patients had single drug substitutions from stavudine to zidovudine and 27 (7%) had single drug substitutions from nevirapine to efavirenz. At baseline, the mean serum lipid concentrations were 120 mg/dL for TC, 53 mg/dL for LDL-c, 29 mg/dL for HDL-c, and 123 mg/dL for TG; values were generally comparable for men and women. During 24 months of treatment, TC increased by a mean of 31 mg/dL, LDL-c by a mean of 26 mg/dL, and HDL-c by a mean of 19 mg/dL, whereas the TC/HDL-c ratio decreased from a mean of 4.6 to 3.4 (all changes, P < 0.001). TG levels initially decreased and then returned to baseline levels by 24 months. At baseline and 24 months, respectively, TC was > or =200 mg/dL for 2% and 10% of patients, LDL-c was > or =130 mg/dL for 1% and 6%, HDL-c was <40 mg/dL for 88% and 41%, and TG were > or =150 mg/dL for 23% and 20%. CONCLUSIONS: Rural Ugandans with advanced HIV disease initiating nevirapine- or efavirenz-based HAART experienced infrequent elevations in TC, LDL-c, and TG at baseline and after 24 months of therapy. Increases in HDL-c levels were substantial and proportionally greater than increases in TC or LDL-c levels. The risk of CVD and how it is affected by lipid changes in this rural African population are unknown. However, the changes we observed after 24 months of HAART seem unlikely to increase the risk of CVD. PMID- 18398973 TI - Nevirapine concentrations in newborns receiving an extended prophylactic regimen. AB - BACKGROUND: The optimal neonatal antiretroviral (ARV) regimen for prevention of HIV mother-to-child transmission (MTCT) is unknown for infants born to mothers who receive no ARVs during pregnancy. METHODS: As part of a protocol comparing the efficacy of 3 neonatal ARV regimens in preventing HIV-1 MTCT in neonates born to mothers who receive no prenatal treatment with ARVs, we devised a 3-dose nevirapine (NVP) regimen with the goal of maintaining the NVP plasma concentration >100 ng/mL (10 times the in vitro median inhibitory concentration of 10 ng/mL) during the first 2 weeks of life. NVP concentrations were measured in 14 newborns participating in a pharmacokinetics substudy during the second week of life and in single samples from 30 more newborns on day 10 to 14. RESULTS: The median NVP elimination half-life was 30.2 hours (range: 17.8 to 50.3 hours). The NVP concentration remained greater than the target of 100 ng/mL in all samples collected through day 10 of life. By day 14, more than half of the newborns in the pharmacokinetic substudy had NVP levels <100 ng/mL, although only 1 neonate had no detectable NVP. CONCLUSION: Although this regimen failed to meet our 100-ng/mL target, it did maintain detectable NVP concentrations in nearly all newborns through the end of the second week of life and is to be used in the parent efficacy protocol. PMID- 18398974 TI - Intermediate highly active antiretroviral therapy adherence thresholds and empirical models for the development of drug resistance mutations. PMID- 18398972 TI - Temporal and lateral dynamics of HIV shedding and elevated sodium in breast milk among HIV-positive mothers during the first 4 months of breast-feeding. AB - OBJECTIVE: To better understand the dynamics of breast milk HIV shedding and its relation to postnatal HIV transmission, we investigated the temporal and lateral relations of breast milk viral shedding and sodium concentrations in HIV-positive women. DESIGN: This was a longitudinal cohort study in Lusaka, Zambia. METHOD: We examined patterns of HIV shedding in breast milk over the first 4 months of breast-feeding and their correlations with postnatal HIV transmission among 138 breast-feeding mothers. Sodium concentration in breast milk was also examined in the same samples and in breast milk from 23 HIV-negative controls. RESULTS: Higher breast milk viral load at 1 week, 1 month, and 4 months and consistent viral shedding in breast milk were significantly associated with increased risk of HIV transmission. Elevated breast milk sodium concentration (> or =13 mmol/L) at 4 months was associated with HIV transmission, low maternal CD4 cell count, and high maternal plasma viral load. Elevated sodium concentration at 1 week postpartum was common and was not associated with any of these parameters. CONCLUSIONS: Consistent viral shedding and high breast milk viral load are strong predictors of mother-to-child HIV transmission. Although sodium concentrations later in breast-feeding correlate with breast milk viral load, increased breast milk sodium is normal in early lactation and does not predict HIV transmission. PMID- 18398975 TI - Similar efficacy of lopinavir/ritonavir-containing regimens among clades B and F HIV-1-Infected individuals in Brazil. PMID- 18398976 TI - Impact of menopause on condom use by HIV-seropositive and comparison seronegative women. PMID- 18398977 TI - Durability of clinical responses to anti-HIV therapies. PMID- 18398978 TI - Policy watch. Encouraging leadership that actually leads us. PMID- 18398979 TI - Osteonecrosis in HIV-infected persons: radiographic findings delay clinical diagnosis. AB - Patients with HIV infection demonstrate an unexpectedly high incidence of bone related disorders, most notably osteonecrosis. We describe 4 HIV-infected patients with osteonecrosis for whom reliance on plain radiographs for establishing the diagnosis was misleading and resulted in a delay in diagnosis. NOne o four patients had significant previously reported risk factors that are associated with osteonecrosis in HIV-infected patients. Osteonecrosis appears to be yet another complication of HIV disease or tis related therapies that has potential for significant morbidity. This disease typically requires surgical intervention for optimal management. Clinicians should maintain a high index of suspicion for osteonecrosis in this patient population in cases of unexplained bone pain or persistent groin pain. Given the high incidence of osteonecrosis in patients with HIV infection, unexplained osteonecrosis should prompt HIV screening, particularly in the absence of identifiable risk factors. PMID- 18398980 TI - Editorial comment: osteonecrosis in HIV-infected persons--does delayed diagnosis matter? PMID- 18398981 TI - Images in HIV/AIDS. Sweet syndrome and syphilis in an HIV-infected person. PMID- 18398982 TI - Editorial comment: Sweet syndrome--a diagnosis seeking a cause. PMID- 18398983 TI - Southwest China opens methadone clinics in border areas to prevent spread of HIV. PMID- 18398984 TI - Preventing AIDS through animation. PMID- 18398985 TI - Current health disparities in HIV/AIDS. AB - The CDC and other public health organizations have identified numerous disparities in the incidence and outcomes of HIV disease among different population groups. Data suggest that some disparities are expected and unavoidable (eg, HIV incidence by age), while others imply inequalities or inequalities that are potentially remediable (eg, differential access by race or sex to HIV prevention and treatment services). Women and minorities, especially those in South and rural areas, have recently been identified as underserved populations at high risk for increased morbidity and mortality from HIV/AIDS. This article reviews current epidemiological trends in HIV/AIDS outcomes, key contributors to observed and emerging health disparities, and strategies that are being employed to overcome important modifiable disparities. PMID- 18398986 TI - Report from the 14th Retrovirus Conference. The HIV/AIDS epidemic in the U.S. PMID- 18398987 TI - Report from the 14th Retrovirus Conference. Antiviral therapy outcomes: large survey highlights differences and similarities worldwide. PMID- 18398988 TI - Report from the 14th Retrovirus Conference. New classes, new agents: changing the paradigm of antiretroviral treatment? PMID- 18398989 TI - Report from the 14th Retrovirus Conference. Metabolic complications: lipoatrophy, lipohypertrophy, and cardiovascular risk. PMID- 18398990 TI - Report from the 14th Retrovirus Conference. New data on HIV and viral hepatitis coinfection. PMID- 18398991 TI - New safety information for erythropoiesis-stimulating agents. PMID- 18398992 TI - Serious non-AIDS conditions: redefining the spectrum of HIV-related disease. PMID- 18398993 TI - FDA approval: maraviroc. PMID- 18398994 TI - Report from the 4th IAS Conference on HIV Pathogenesis, Treatment and Prevention. HLA B-5701 and abacavir hypersensitivity. PMID- 18398995 TI - Report from the 4th IAS Conference on HIV Pathogenesis, Treatment and Prevention. When to start HIV treatment in infants. PMID- 18398996 TI - Report from the 4th IAS Conference on HIV Pathogenesis, Treatment and Prevention. Further exploring the findings from SMART. PMID- 18398997 TI - Report from the 4th IAS Conference on HIV Pathogenesis, Treatment and Prevention. Acyclovir to prevent HIV infection? PMID- 18398998 TI - Report from the 4th IAS Conference on HIV Pathogenesis, Treatment and Prevention. Disappointing data from anti-HIV microbicide trials. PMID- 18398999 TI - Report from the 9th International Workshop on Adverse Drug Reactions and Lipodystrophy in HIV. PMID- 18399000 TI - HIV-1 vaccine candidate ineffective. PMID- 18399001 TI - Caution regarding nelfinavir use in children and pregnant women. AB - The discover of a potential carcinogen in nelfinavir has prompted new recommendations regarding use of this drug. PMID- 18399002 TI - Report from the 47th Interscience Conference on Antimicrobial Agents and Chemotherapy. PMID- 18399003 TI - Raltegravir, a new HIV integrase inhibitor. AB - Because raltegravir inhibits a novel viral target, even HIV that is resistant to other antiretroviral-medication classes is sensitive to this drug. PMID- 18399004 TI - Lower dose of ritonavir approved for use with fosamprenavir. PMID- 18399005 TI - Enfuvirtide and Biojector 2000 needle-free injection device. PMID- 18399006 TI - Top stories of 2007. PMID- 18399007 TI - HIV-1 vaccine might increase infection risk in certain subgroups. PMID- 18399008 TI - Updated antiretroviral treatment guidelines from DHHS and EACS. PMID- 18399009 TI - Crystal methamphetamine and HIV/AIDS. PMID- 18399010 TI - Revised guidelines for antiretroviral use in pregnant women. PMID- 18399012 TI - FDA approval: etravirine. PMID- 18399011 TI - Updated guideline for occupational postexposure prophylaxis for HIV. PMID- 18399013 TI - DHHS treatment guidelines tweaked again. PMID- 18399015 TI - Is liquid-based cytology better than Pap tests for CIN 2? PMID- 18399014 TI - Revised package insert for atazanavir. PMID- 18399016 TI - Should you delay solid food in infants to prevent atopy? PMID- 18399017 TI - Does adding vitamin K to warfarin improve anticoagulation control? PMID- 18399018 TI - Should you screen for asymptomatic carotid artery stenosis? PMID- 18399019 TI - Refreshing contradictions and lost in classification. PMID- 18399020 TI - Is Zetia a 'do nothing' drug? PMID- 18399021 TI - GPs' and health visitors' views on the diagnosis and management of postnatal depression: a qualitative study. AB - BACKGROUND: In the UK, 8-15% of women suffer from postnatal depression, with long term consequences for maternal mood and child development. Previous literature suggests that health visitors struggle with their conflicting roles with respect to mother and infant. Current policy is redirecting the emphasis and organisation of health visitor work, but guidelines state that health visitors and GPs should continue to have a major role in the detection and management of postnatal depression. AIM: To explore the views of GPs and health visitors on the diagnosis and management of postnatal depression. DESIGN OF STUDY: A qualitative study nested within a multicentre randomised controlled trial. SETTING: Nine primary care trusts in Bristol, Manchester, and London. METHOD: In-depth interviews with GPs and health visitors from primary care trusts participating in a randomised controlled trial of antidepressants versus health visitor-delivered non-directive counselling. Interviews were audiotaped and fully transcribed. Thematic analysis with an iterative approach was used to develop conceptual categories from the transcripts. RESULTS: Nineteen GPs and 14 health visitors were interviewed. GPs and health visitors described their work in making and negotiating the diagnosis of postnatal depression, the value of a long-term relationship with the woman, and how labelling affects management of women with postnatal depression. Responders described how they viewed others' roles in the management of postnatal depression, and how national policy and local organisational changes had an impact on patient care, so that no one health professional was assuming overall responsibility for the care of women with postnatal depression. CONCLUSION: Ongoing organisational changes within primary care, such as the implementation of corporate working by health visitors, affect care provided to women after birth, which in turn has an impact on the diagnosis and management of postnatal depression. PMID- 18399022 TI - Feasibility of an exercise intervention for women with postnatal depression: a pilot randomised controlled trial. AB - BACKGROUND: Postnatal depression is a serious mental health problem that may be reduced by exercise. AIM: This study examined the feasibility of an exercise intervention for women with postnatal depression, and assessed which methods of recruitment are most effective. DESIGN OF STUDY: Randomised controlled trial. SETTING: General practice and the community. METHOD: Participants were recruited from various sources and randomised to an exercise intervention or usual care with follow-up at 12 weeks. As well as assessing feasibility, other trial outcomes included exercise participation and self-efficacy for exercise. Levels of depression were assessed but the study was not powered to show a difference in this. RESULTS: The recruitment rate of eligible patients was 23.1%. The highest recruitment rate was via referral from the psychiatric mother and baby unit (9/28; 32.1%), followed by invitation letters from GPs (24/93; 25.8%). Thirty eight eligible participants were randomised. At follow-up there was no significant difference in exercise participation between groups. The intervention group reported significantly higher self-efficacy for exercise compared to usual care, but depression scores did not differ. CONCLUSION: Exercise participation over the 12-week period was not significantly increased, possibly because it is difficult to motivate women with postnatal depression to exercise, or the intervention was not sufficiently intensive. Eligible patients were recruited into this study but response rates were low. Optimum methods of recruitment in this difficult-to-reach population are required prior to a substantive trial. Further research is imperative given poorly-evidenced recommendations by the National Institute for Health and Clinical Excellence to consider this treatment. PMID- 18399023 TI - Medicine Meets Virtual Reality 16. Parallel, combinatorial, convergent: NextMed by Design. Preface. PMID- 18399024 TI - [Brief letter: "Gestational diabetes under clinical conditions in aspired normoglycemia: investigation for correlation of blood glucose daily profiles and fetometric ultrasound parameters"]. PMID- 18399025 TI - Evolution of the ISSCR Industry Committee. PMID- 18399026 TI - Biopolis: village on a hill. PMID- 18399027 TI - Stem cells down under-ISSCR 2007. AB - Cairns, Queensland, Australia provided the venue for the 5th annual meeting of the International Society for Stem Cell Research (ISSCR), June 17-20, 2007. Consonant with the young society's mission to serve the international stem cell community, this meeting was the first held outside North America. The meeting drew attendees from 44 countries, with excellent representation from the Asia/Pacific region. The more than 120 presentations and 1000 posters covered virtually all aspects of the stem cell field and provided a snapshot of current areas of investigation. Here we review some of the newest findings in an effort to convey the energy and excitement of the meeting and the tempo of stem cell science. PMID- 18399028 TI - [Off-label applications (professional rationales, financial possibilities)]. PMID- 18399029 TI - [A naturalistic, observational study of outpatients with schizophrenia: efficacy and safety results after 6 months. The International Schizophrenia Outpatient Health Outcomes study, IC-SOHO]. AB - The International Schizophrenia Outpatient Health Outcomes study, IC-SOHO is a three-year international observational study that investigates clinical and health outcomes of antipsychotic treatments. 7658 outpatients treated for schizophrenia were enrolled in the study, who needed an antipsychotic therapy to initiate or switch. The primary analysis compared the group taking olanzapine with the group taking any other antipsychotics, while the secondary comparison was performed between those treated with olanzapine and those with risperidone. Efficacy analysis was carried out based on changes in Clinical Global Impression of Severity scale (CGI-S), which was performed at a global symptom level, as well as with respect to the patients' positive, negative, cognitive and depressive symptoms. In addition, adverse events were also evaluated. Results of the analysis of the 3- and 6-month data from Hungary are disclosed in this publication. 200 patients were enrolled in the country. Demographics of the treatment groups were not significantly different. At 3 and 6 months after treatment initiation, there were no significant between-group differences in improvement of global symptomatology, however, cognitive symptoms improved more in the Olanzapine-group compared to those taking other antipsychotics (p<0.05). In patients showing Extrapyramidal Symptoms (EPS) at baseline, these symptoms finished to a greater extent among those receiving olanzapine than in those receiving other antipsychotics (after 6 months D<0.0001). Half a year later, significantly less patients showed extrapyramidal adverse events (p=0,0007), and the previous EPS terminated to a greater extent (p=0.0016) in the olanzapine group, as compared to those taking risperidone. No between-group differences were found in changes of sexual functions, as well as of weight and Body Mass Index measures. Switching antipsychotic initiated at study baseline, and adding-on one or more other antipsychotic to the initial one, were significantly less frequent in the Olanzapine-group compared to those initiated other antipsychotics. In the first 3 months, treatment compliance was significantly higher with olanzapine therapy than with other antipsychotic treatments, and with risperidone respectively. Results from the Hungarian sample correspond with results from higher analysis levels of wider patient populations of IC-SOHO study. Olanzapine showed outstanding efficacy in lessening cognitive disturbances and global clinical symptomatology associated with schizophrenia. Extrapyramidal symptoms of patients treated with olanzapine improved significantly better compared with those patients who received other antipsychotics and risperidone respectively. PMID- 18399030 TI - Comparison of propofol and etomidate regarding impact on seizure threshold during electroconvulsive therapy in patients with schizophrenia. AB - BACKGROUND: While propofol is known to shorten seizures during electroconvulsive therapy, in our previous study on patients with schizophrenia, there was no need for more frequent restimulations when using propofol compared with etomidate. We hypothesized that etomidate and propofol have similar effects on seizure activity in cases where seizure duration is shorter than 20 seconds. In this study, etomidate and propofol are compared regarding their impact on seizure threshold and seizure duration. METHOD: 30 schizophrenic patients participated in this prospective randomized cross-over study. For anesthetic induction were 1 mg/kg of propofol or 0.2 mg/kg of etomidate used alternately. For both anesthetics, seizure threshold was determined by titrating the dose of the stimulus necessary for eliciting a seizure. Seizure durations were also compared. RESULTS: After etomidate induction, seizure durations registered either by EEG or by EMG were longer than propofol treated cases (EEG: 49.6+/-23.1 s, versus 39.7+/-19 s, p=0.026; EMG: 41.4+/-22 s, versus 32.8+/-17.6 s, p=0.016). However, no significant differences were found for minimum seizure eliciting stimulation energy or the number of restimulations between the two anesthetics (41.58 mC+/ 13.6 mC, versus 41.58 mC+/-11.1 mC, p=1.00). CONCLUSION: During the ECT of patients with schizophrenia, propofol was shown to possess significant seizure shortening properties, but it does not elevate seizure threshold or drop seizure duration under the minimal threshold more frequently than etomidate does. Based on these findings, we conclude that the use of propofol does not result in a greater electric load on the patients than etomidate. PMID- 18399031 TI - Enhancing effect of mirtazapine on cognitive functions associated with prefrontal cortex in patients with recurrent depression. AB - INTRODUCTION: We put forward a hypothesis that a therapeutic administration of mirtazapine to depressed patients, due to pharmacological profile of the drug, could enhance cognitive functions associated with prefrontal cortex activity. METHODS: The study was performed on depressed patients receiving mirtazapine for the period of 6 months. Neuropsychological assessments after 3 and 6 months of treatment were performed by the Wisconsin Card Sorting Test, N-back test, TMT and Stroop tests. RESULTS: During acute depressive episode, a significant impairment on all neuropsychological tests was evident. Substantial improvement in performance has been noted after 3 and 6 months of mirtazapine treatment, and, after 6 months, a majority of the investigated patients achieved the results within the range of matched healthy control subjects. Improvement on neuropsychological tests after treatment with mirtazapine showed no correlation with the degree of amelioration of depression. DISCUSSION: Mirtazapine may exert a favorable influence on cognitive functions associated with prefrontal cortex in depressed patients. The lack of direct correlation with improvement of depressive symptoms suggests that mirtazapine may possess specific pro-cognitive properties. PMID- 18399032 TI - [The medications from the viewpoint of the psychotic patients]. AB - In this paper is given a short introduction to the functioning of the group therapy of psychotic patients at our unit; outlining also other aspects of their treatment (medication, hospitalisation, group therapy). By collecting examples from the reports of the group therapy we illustrate how the medicaments appear in the conversations. We analyse the given data from three different point of view: interactional, thematic and by the psychotic contents. In the quotations first we can find examples of the positive and negative attitudes towards medication, injections. Then patients speak about the states when medicament gives no more help, about their imaginations of alternative ways of healing, finally we can follow how the topic of medicaments mix with their delusions. PMID- 18399033 TI - [Measuring the cognitive deficit associated with schizophrenia: presentation of the MATRICS neuropsychological test battery]. AB - The aim of the present study is to introduce and give an overview of the Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS) neurocognitive test battery, which has been recently developed in the United States specifically for the assessment of neurocognitive deficits in patients with schizophrenia. The MATRICS test battery consists of 10 individually administered tests to measure cognitive performance in the following 7 domains: speed of processing, attention/vigilance, working memory, verbal learning, visual learning, reasoning and problem solving, and social cognition. Besides giving a detailed description of the test, the authors also aim to point out the advantages and novel features of the test compared to already existing neuropsychological test batteries used to assess cognition in patients with schizophrenia. The future applicability of the MATRICS test battery will be related to current trends in clinical trials and pharmacological studies. The significance of the standardization and adaptation of the test to Hungarian is emphasized and future possibilities of applying the test in a Hungarian language environment discussed. PMID- 18399034 TI - [Acquired drives. The cortical mechanism responsible to the emergence and development of social existence]. AB - This paper is a brief interpretation of the theory (J. Knoll: The Brain and Its Self, Springer, 2005) the main message of which is that the appearance of the mammalian brain with the ability to acquire drives ensured the development of social life, and eventually led to the evolution of the human society. In the mammalian brain capable to acquire drives, untrained cortical neurons (Group 1) possess the potentiality to change their functional state in response to practice, training, or experience in three consecutive stages, namely, by getting involved in (a) an extinguishable conditioned reflex (ECR) (Group 2), (b) an inextinguishable conditioned reflex (ICR) (Group 3), or (c)an acquired drive (Group 4). The activity of the cortical neurons belonging to Group 3 and 4 is inseparable from conscious perception. In any moment of life self is the sum of those cortical neurons that have already changed their functional significance and belong to Group 3 or 4. Metaphorically, every human being is born with a telencephalon that resembles a book with over 100 billion empty pages (untrained, naive cortical neurons, Group 1), and with the capacity to inscribe as much as possible in this book throughout life. Whenever a drive is acquired, chains of ICRs are fixed, neurons responsible for emotions are also coupled to the integral whole, thus cognitive/volitional consciousness is necessarily inseparable from an affective state of consciousness. Cortical neurons belonging to Group 3 or 4 continuously synthesize their specific enhancer substance within their capacity. This means that even in the vigilant resting state (leisure), in the absence of a dominant drive, as well as in the non-vigilant resting state (sleeping), the cortical neurons representing the totality of the already fixed ICRs and acquired drives are permanently under the influence of their specific enhancer substance. Although the level of this permanent, undulating activation remains low, it is unpredictable as to when any group of cortical neurons will be influenced by enhancer substances on the level already inseparable from conscious perception. Thus, as the totality of the cortical neurons belonging to Group 3 or 4 works continuously on an unconscious level, there is a steadily operating, chaotic background noise in the human telencephalon. Even in the active state ("fight or flight" behavior, goal-seeking), when the actually dominant drive determines the rational goal to be reached, the noise is suppressed, but cannot cease to exist. But it never endangers the function of the actually dominant innate or acquired drive. From this situation it follows that the rational brain activity is necessarily amalgamated with an irrational brain activity and we live through every moment of our life experiencing the totality of order and chaos in our brain. Human society the maintenance of which has always required the proper manipulation of the brain of its members still finds itself in a state of development. It seeks its final equilibrium: namely, that state in which behavioral modification induced by the home/school/society triad will be based, from birth until death, on the exact knowledge of the natural laws that keep the brain and its self going. In this way, members of the community will understand that simultaneity of order and chaos in their brain is the physiological reality that determines human activity, and will consciously try to find the acquired drives that optimally fit their natural endowments. For the time being those who have been lucky enough to acquire the best fitting drives in due time, in the early uphill period of life, have had fair chances for success and happiness. In contrast, those who for any reason have missed this opportunity will remain frustrated and look for 'ersatz'. It seems reasonable to conclude that order and chaos are of equal importance in our brain. Without the ability to adapt ourselves to the concrete (science), we would not be able to survive; without the ability which allows detachment from the concrete and explorations in the infinite (art), life would not be worth living. Thus, the human society, this most sophisticated form of organized life on earth is still in trial and error phase of its development. It seeks to outgrow the myth-directed era of its history and come to its final state, the reason-directed human society. PMID- 18399035 TI - Memory and schizophrenia. PMID- 18399036 TI - Functional mechanisms of episodic memory impairment in schizophrenia. AB - OBJECTIVE: To achieve a better understanding of the functional mechanisms underlying episodic memory dysfunction in schizophrenia, which is a prerequisite for unravelling schizophrenia's neural correlates in neuroimaging studies and, more generally, for developing an integrated approach to the pathophysiology of schizophrenia. It is also crucial for developing cognitive remediation. METHOD: This paper reviews empirical evidence of episodic memory dysfunction in schizophrenia obtained with reference to various theoretical models of episodic memory. RESULTS: All the studies converge to show a significant impairment of the critical feature of episodic memory: conscious recollection. Schizophrenia is also associated with a defect of autobiographical memory. The episodic memory dysfunction results from a predominant failure of strategic processing at encoding, although an impairment of strategic processing at retrieval cannot be ruled out. The possibility that it is not the execution of the encoding strategies that is defective but, rather, their self-initiation by the patients is plausible. CONCLUSIONS: These findings may explain some behavioural abnormalities associated with schizophrenia, notably, inadequate functional outcomes in everyday life. They may also have implications for cognitive remediation and better social and work functioning of patients with schizophrenia. PMID- 18399037 TI - Episodic memory bias and the symptoms of schizophrenia. AB - Much of the research on episodic memory in schizophrenia spectrum disorders has focused on memory deficits and how they relate to clinical measures such as outcome. Memory bias refers to the modulatory influence that state or trait psychopathology may exert on memory performance for specific categories of stimuli, often emotional in nature. For example, subjects suffering from depression frequently have better memory for negative stimuli than for neutral or positive ones. This dimension of memory function has received only scant attention in schizophrenia research but could provide fresh new insights into the relation between symptoms and neurocognition. This paper reviews the studies that have explored memory biases in individuals with schizophrenia. With respect to positive symptoms, we examine studies that have explored the link between persecutory delusions and memory bias for threatening information and between psychosis and a memory bias toward external source memory. Although relatively few studies have examined negative symptoms, we also review preliminary evidence indicating that flat affect and anhedonia may lead to some specific emotional memory biases. Finally, we present recent findings from our group delineating the relation between emotional valence for faces and memory bias toward novelty and familiarity, both in schizophrenia patients and in healthy control subjects. A better understanding of the biasing effects of psychopathology on memory in schizophrenia (but also on other cognitive functions, such as attention, attribution, and so forth) may provide a stronger association between positive and negative symptoms and memory function. Memory measures sensitive to such biases may turn out to be stronger predictors of clinical and functional outcome. PMID- 18399038 TI - A national survey of Canadian psychiatry residents' perceptions of psychotherapy training. AB - OBJECTIVE: To explore Canadian psychiatry residents' perceptions of their psychotherapy training and identify factors that may influence decisions to practise psychotherapy after graduation. METHOD: We surveyed psychiatry residents at all training sites across Canada, using a self-report questionnaire. RESULTS: The response rate was 63%. Of the respondents, 68% indicated that the prospect of learning and practising psychotherapy was a factor in their decisions to become psychiatrists, and 87% considered their ability to practise psychotherapy to be important to their identities as psychiatrists. The majority of residents (71%) were generally satisfied with their psychotherapy training. Among the graduating class of residents, 84% anticipate practising psychotherapy in some capacity. Satisfaction with their overall training experience and supervision and feeling competent to perform psychotherapy were significantly associated with their decisions to practise psychotherapy after graduation. CONCLUSIONS: Most psychiatry residents currently enrolled in postgraduate training programs across Canada view psychotherapy as having an important role in the way they anticipate practising psychiatry. PMID- 18399039 TI - Combined therapy of major depression with concomitant borderline personality disorder: comparison of interpersonal and cognitive psychotherapy. AB - OBJECTIVE: The combination of antidepressants and brief psychotherapies has been proven more efficacious in treating major depression and is particularly recommended in patients with concomitant personality disorders. We compare the effects of 2 combined therapies, fluoxetine and interpersonal therapy (IPT) or fluoxetine and cognitive therapy (CT), on major depression in patients with borderline personality disorder (BPD). METHOD: Thirty-five consecutive outpatients with a diagnosis of BPD and a major depressive episode (not bipolar and not psychotic) were enrolled. They were randomly assigned to 1 of the 2 combined treatments and treated for 24 weeks. Assessment included a semistructured interview, Clinical Global Impression (CGI) scale, Hamilton Depression Rating Scale (HDRS), Hamilton Anxiety Rating Scale (HARS), Beck Depression Inventory-II (BDI-II), Social and Occupational Functioning Assessment Scale (SOFAS), Satisfaction Profile (SAT-P) for quality of life (QOL), and Inventory of Interpersonal Problems (IIP-64). Statistical analysis was performed using the univariate General Linear Model to calculate the effects of duration and type of treatment. RESULTS: No significant differences between treatments were found at CGI, HDRS, BDI-II, and SOFAS score. Combined treatment with CT had greater effects on HARS score and on psychological functioning factor of SAT-P. Combined treatment with IPT was more effective on social functioning factor of SAT-P and on domains domineering or controlling and intrusive or needy of IIP-64. CONCLUSIONS: Both combined therapies are efficacious in treating major depression in patients with BPD. Differences between CT and IPT concern specific features of subjective QOL and interpersonal problems. These findings lack reliable comparisons and need to be replicated. PMID- 18399040 TI - Neural representation of maternal face processing: a functional magnetic resonance imaging study. AB - OBJECTIVE: The mother-child relationship may have important implications for emotional development and adult psychopathology. The objective of this study was to examine brain responses to processing maternal faces in healthy adult women. METHODS: Ten healthy adult female volunteers with adequate early-life maternal care and a normal relationship with their living mothers participated in the study. Functional magnetic resonance imaging was used to examine brain responses to pictures of the subject's mother, a close female friend, and 2 age-matched female strangers during passive viewing, valence (emotional), and salience (self relevance) evaluations. RESULTS: The main contrast of mother, compared with all others (that is, friend and older and younger strangers), demonstrated the following: first, significant activation in the left posterior cingulate cortex precuneus (PCC-Pcu), collapsed across all tasks; second, right ventromedial prefrontal cortex-anterior cingulate cortex (VMPFC-ACC) activation during the valence condition; and third, left dorsolateral prefrontal cortex (DLPFC) activation during the salience condition. In the region-of-interest analyses, the VMPFC-ACC and DLPFC showed significant activations in response to mothers' faces and deactivation in response to control faces. Among the 3 regions, only VMPFC ACC activity distinguished the unique processing of one's own mother's face from that of a close friend. PCC-Pcu activations demonstrate a graded response (mother > friend > strangers) and, further, demonstrated differential response with respect to mothering style. CONCLUSIONS: The activation in prefrontal and cingulate cortices related to maternal face processing is consistent with their implicated roles in mother-infant interactions, personal familiarity, and emotional and self-relevant processing. These findings suggest a neural basis for maternal attachment and propose a focus for future studies aimed at investigating the impact of disrupted maternal attachment on emotional development. PMID- 18399041 TI - [Treatment progress scale for violent psychosis patients]. AB - OBJECTIVES: The dangerousness of psychiatric patients is related to many well documented factors in literature. To better document the course of dangerousness in most violent psychotic patients with severe and persistent diseases, we have developed a Treatment Progress Scale (TPS). METHOD: The TPS was developed based on the literature research and the expertise of a multidisciplinary team. RESULTS: The instrument has good interrater reliability and has shown to be easy to use, after being implemented for 5 years in a treatment unit at Philippe-Pinel Institute. The instrument provides a systematic assessment of important clinical parameters validating treatment observance and patient evolution over months. It also builds on the daily observations made by different members of the multidisciplinary team, the common language of patients and staff, the transparency of our work with patients, and the clear identification of most treatment targets. It also brings a valued complement to the dangerousness assessment. CONCLUSIONS: The TPS is a relevant instrument for the violent psychotic inpatient specific population. It provides a better identification of treatment progress and helps to specify dangerousness from an evolution perspective. Variations may be developed to assess outpatients and patients in a general setting. PMID- 18399042 TI - Depressed mood in the suicidal population. AB - OBJECTIVE: To determine whether, and to what extent, those who reported suicidal ideation and (or) attempts in the previous year, without meeting the criteria for major depressive episode (MDE), might still have experienced subthreshold depression. METHOD: Data from the Canadian Community Health Survey: Mental Health and Well-Being (CCHS 1.2) were examined to estimate and compare the prevalence of depressed mood, a core symptom of MDE and subthreshold depression, within the previous month for the following 4 population groups: MDE within the previous month [D(1 month)], MDE within the previous year but not the previous month [D(1 year)], suicidality without MDE within the previous year (D-S+), and no MDE or suicidality within the previous year (D-S-). RESULTS: As expected, the D(1 month) group showed the highest prevalence of depressed mood in the previous month. However, the D-S+ group had a significantly higher prevalence of depressed mood than the D-S- group and only a slightly lower level than that of the D(1 year) group. CONCLUSIONS: A fraction of those who reported suicidality in the previous year met the criteria for MDE during that time; however, subthreshold depression might be common in the suicidal population. Future epidemiologic research in the areas of suicidality and depression will benefit from a comprehensive assessment of subthreshold depression. PMID- 18399043 TI - Re: In debate: are antidepressants as effective as claimed? PMID- 18399044 TI - Effectiveness of a computer skills program to improve written communication in children with Developmental Coordination Disorder. AB - Word processing using a keyboard is an option for children with Developmental Coordination Disorder (DCD) who have printing/ handwriting difficulties. A five replication single-subject ABA design was used to explore the ability of 6 children with DCD (entering Grades 3-5) experiencing problems with printing/handwriting to learn computer skills. Following a two-week intervention all children were familiar with basic computer functions and showed improved word processing skills. Touch-keyboarding speed increased for the five children entering Grades 4-5, but only one child acquired a speed (letters/minute) comparable to printing/ handwriting. Four children demonstrated increased text production when generating a story, but none achieved speeds comparable to printing/handwriting. Five-finger touch-keyboarding proficiency was better for children in Grades 4-5. Therapists observed improved legibility and increased motivation. Results suggest that the child's grade, program support, and program length should be considered when making decisions about keyboarding. PMID- 18399045 TI - Physical activity of young children: a two-year follow-up. AB - Inadequate physical activity in children is a major health concern. The purpose of this study was to examine changes in physical activity of boys and girls, between 6-8 and 8-10 years of age and how activity patterns correlated with selected family, child, and environment factors. The sample included 59 children without motor delays (26 boys and 23 girls) between 8 and 10 years of age. Twenty two of the children participated in a previous study at 6-8 years of age. Parents completed a questionnaire on their children's non-physical and physical activities. Children wore a pedometer during two weekdays and two weekend days. The results indicate that girls spent more time on homework and reading and on crafts and indoor play than boys. Girls spent more time on musical and cultural activities and boys spent more time on screen-based activities at 8-10 years of age. Children spent significantly less time on physical activity at 8-10 years of age. Boys took more steps per day than girls on weekends. The average number of steps taken per weekday increased for boys, but not girls, at 8-10 years of age. There was an inverse relationship between body mass index and number of steps taken per day (weekdays r = -.28; weekend r = -.32). Socioeconomic status was associated with the number of steps taken by children on weekends (r = .34). The results have implications for physical activities for girls and school and community programs for children. PMID- 18399046 TI - Self-reported quality of life in adolescents with cerebral palsy. AB - Health Related Quality of Life (HRQOL) is an important outcome in the delivery of health care. Research on the HRQOL in young people with cerebral palsy (CP) has relied on proxy-reports from parents. The purpose of this study was to investigate the self-reported HRQOL of young people with CP. A survey was mailed to 229 adolescents with CP in South Australia, of which 118 responded (51.5%). Seventy-one participants 11 to 17 years of age, self-reported HRQOL on the Pediatric Quality of Life Inventory 4.0 (PedsQL). Thirty-eight youth were deemed by their parent/guardian as having insufficient cognitive ability to self-report HRQOL and nine respondents returned their surveys incomplete. The mean PedsQL Physical Function score was 57.3 (SD = 24.3), the mean Psychosocial Function score was 64.5 (SD = 15.9), and the Overall PedsQL score was 62.0 (SD = 16.7). Compared to norms for children without disabilities, 67% of participants had an Overall PedsQL score greater than 1 SD below the mean. PedsQL scores were related to gross motor function classification level (Spearman's rho = -0.54), number of health issues (rho = -0.51), and socioeconomic status (rho = 0.28), but not age, gender, quality of sleep, or whether parent assistance was needed to complete the PedsQL. The results have implications for policy and efforts to identify and address barriers to full and satisfying participation in mainstream schools and community activities. PMID- 18399047 TI - Item test-retest reliability and responsiveness of the School Outcomes Measure (SOM). AB - The School Outcomes Measure (SOM) is a minimal data set designed to measure outcomes of students who receive school-based occupational therapy and physical therapy. The purpose of this study was to determine item test-retest reliability and the responsiveness to change of the SOM. Thirty-two occupational therapists and physical therapists collected data on 73 students, age 3-21 years, stratified into two groups based on the Gross Motor Function Classification System: those with mild/moderate functional limitations (Levels I, II, III) and those with severe functional limitations (Levels IV, V). Weighted kappa statistics (0.68 1.0) indicate good to excellent item test-retest reliability. Repeated measures analysis of variance indicated that the SOM is responsive to change in children with mild/moderate functional limitations but is less responsive to change in children with severe functional limitations. The findings support the use of the SOM to measure outcomes for students with mild/moderate functional limitations as well as enabling program evaluation of therapy services provided in school settings. More research is needed to evaluate responsiveness in students with severe functional limitations. PMID- 18399048 TI - Developmental trajectories and correlates of sensory processing in young boys with fragile X syndrome. AB - BACKGROUND AND PURPOSE: No longitudinal study on sensory processing in children with fragile X syndrome (FXS) exists. This study examined developmental trajectories and correlates of sensory processing from infancy through preschool years in 13 boys with FXS. METHOD: Participants were assessed using observational and parent-report measures 2-6 times between 9 and 54 months of age. RESULTS: Over time, an increasing proportion of boys displayed sensory processing that differed significantly from test norms. Observational measures were more sensitive than parent-reports early in infancy. Age and developmental quotient significantly predicted levels of hyporesponsiveness; there was a trend for hyperresponsiveness to increase with age. Baseline physiological and biological measures were not predictive. CONCLUSIONS: Sensory processing problems are observable early and grow increasingly problematic from infancy through the preschool ages. Early identification and intervention may attenuate long-term difficulties for children with FXS. PMID- 18399049 TI - Changing your practice. PMID- 18399052 TI - Would you tell the parents? PMID- 18399053 TI - Beyond preeclampsia: HELLP syndrome. PMID- 18399054 TI - Cardiac care: managing postoperative bleeding. PMID- 18399055 TI - Nursing shortage: looking to the future. PMID- 18399056 TI - Money doesn't grow on trees. PMID- 18399057 TI - The other end of the needle. PMID- 18399058 TI - Mammographic density--a risk factor for breast cancer. AB - The mammographic density belongs to risk factors for breast cancer. The increased density enhances the risk of interval breast cancers and it also increases the number of false negative as well as false positive results of the mammography. The increase in the mammographic density during the hormone replacement therapy is not identical with the enhanced mammographic density, which is a risk factor for the breast cancer. The mammographic density associated with the hormonal treatment recedes within 14 days after its withdrawal. The high mammographic density is not contraindication of the hormonal treatment, but it results in a preference of tibolone and estrogen replacement therapy over the estrogen gestagen treatment. PMID- 18399059 TI - Genetic polymorphisms as a risk factor for anorexia nervosa. AB - Anorexia nervosa (AN) is an eating disorder affecting mostly young people which could lead to serious complications and consequences. There are ethnical and gender differences in the incidence and prevalence of AN, but the influence of urbanization has not yet been proved. The relationship of genetic background to the risk of AN is still being investigated. In this review we summarize current knowledge about the relationship between AN and polymorphism of substances known to be regulating eating behaviour or metabolic pathways e.g. serotonin, ghrelin, catechol-O-methyl transferase, neuropeptide Y, brain-derived neurotrophic factor and adipokines. PMID- 18399060 TI - Weight, body composition and fat distribution changes of Czech women in the different reproductive phases: a longitudinal study. AB - Women tend to gain weight with age, especially fat mass which shows also regional changes. A cross-sectional study was done on 213 Czech women with the conclusion that there is a progressive weight increase up to the menopause, an increase in absolute and relative fat contribution and fat centralisation up to the postmenopause and these changes seemed to occur even before the weight increase occurs. The same cohort was examined the same way after 3 years for further analysis. The aim was the assessment of weight gain, body composition and fat distribution changes in four age groups representing the reproductive phases in Czech women in 3-year period. 146 healthy Czech women aged 20-65 were classified according to four reproductive phases: fully reproductive women (n=34, mean age 26.96, SD 4.47), pre-menopausal women (n=34, mean age 42.23, SD 2.78), menopausal women (n=45, mean age 51.56, SD 2.61) and postmenopausal women (n=33, mean age 59.55, SD 2.82). Body weight, body composition and fat distribution were determined using classical anthropometric methods in 2000 and 2003. BMI increased significantly in all the groups except for the premenopausal group and was the greatest in the menopausal group. Increase in fat percentage was significant in the fully reproductive (p < 0.001) and menopausal women (p < 0.001), there was no change in the postmenopausal group. Waist, WHR, hip and subgluteal thigh circumference increase significantly in the menopausal group (p < 0.01, p < 0.05, p < 0.05, p < 0.001 respectively). The highest mean values of waist, WHR and even abdominal circumference remain in the postmenopausal group. Changes of all 14 skinfolds and the sum of the peripheral and central skinfolds are shown; the sum of peripheral skinfolds shows the same values at the end of the study while the sum of central skinfolds increases from the fully reproductive to the postmenopausal women. These results permit us to state the following conclusions: the greatest weight gain in the menopausal group suggests weight gain acceleration around menopause. Body fat mass increase terminates in the early postmenopause. Fat centralisation was proved in the menopausal women with still preserved fat deposition in the gluteofemoral area, which was also apparent in the postmenopausal group, however, the postmenopausal women show the highest values of central fat indicators. PMID- 18399061 TI - Sensitivity and specificity of combined perimetric and volumetric evaluations in the diagnosis of arm lymphedema. AB - The objective of the current study was to evaluate the sensitivity and the specificity of perimetry combined with volumetry in the treatment of lymphedema. Ninety women, who had been submitted to breast cancer surgery, were randomly selected in the Government Healthcare Clinic for this study. Only patients who underwent surgical treatment of breast cancer with some degree of lymphadenectomy were included in the study cohort. Individuals with active disease, whether local or otherwise, functional alterations of the upper limbs before breast cancer surgery were not included. The following possibilities were considered: 1--the perimetry evaluation was considered positive when the difference between the affected and unaffected sides was > or =2 cm for any one of the seven measurements and volumetry was > or = 100 mL; 2--perimetry > or =2 cm and volumetry > or =200 mL; 3--a difference > 10% between the two limbs in volumetry and perimetry. Prevalence, sensitivity, specificity, positive predictive value, negative predictive value and accuracy were evaluated statistically with an alpha error of 5% considered acceptable (p-value < 0.05). The mean age of the women was 54.8 +/- 11.7 years. The sensitivity, negative predictive value and accuracy were higher using perimetry when a volume > or =2 cm were considered. The specificity and positive predictive values were better when the difference was greater than 10% between the two limbs by both perimetry and volumetry. Perimetry is a reliable method in the diagnosis of lymphedema when differences > or =2 cm between the two limbs should be considered. PMID- 18399062 TI - Frequency of micronuclei in Mexicans with type 2 diabetes mellitus. AB - A case-control study was carried out on a sample of 15 Mexican patients (40-56 years old) with type 2 diabetes mellitus (DM2) that had developed five years and been treated with oral hypoglycemic drugs (sulfonylurea and/or metformin), with no microvascular or macrovascular complications. The aim of this study was to assess whether Mexican patients with DM2 differed from a control group in the frequency of micronuclei (MN). A control group of 10 individuals without DM2 (38 54 years old) was included. The frequency of MN in binucleated lymphocytes was analyzed according to the Fenech criteria. At time being this investigation should be considered as a preliminary study in which the influence of potential confounders cannot be adequately assessed. However, our result showed a MN frequency significant increase in DM2 patients (6.53 +/- 2.03 per 1000 cells) relative to that of the control group (3.10 +/- 1.79 per 1000 cells). MN may constitute a possible component of a panel of biomarkers for the risk of DM2. This cytogenetic damage also indicates an enhanced risk of cancer, as has been found in previous studies. These results should be validated by other researchers. PMID- 18399063 TI - Complicated decisionmaking in indications for auditory brainstem implant (ABI) in a patient with neurofibromatosis 2. AB - A case of an accidental finding of neurofibromatosis 2 in a practically asymptomatic patient is described. Various therapeutic modalities, including restoration of hearing after vestibular schwannoma surgery with an auditory brainstem implant (ABI), are considered. PMID- 18399064 TI - X-linked dominant chondrodysplasia punctata (CDPX2): multisystemic impact of the defect in cholesterol biosynthesis. AB - Chondrodysplasia punctata represents clinically and genetically a heterogeneous group of disorders characterized by the presence of multiple congenital anomalies and stippled epiphyses. We present clinical course of the disease and the results of metabolic, X-ray and molecular analyses in 19-months old girl with X-linked dominant chondrodysplasia punctata with intrauterine growth retardation, craniofacial dysmorphy, cataracts, cutaneous anomalies including ichthyosis, asymmetric rhizomesomelic shortness of the limbs, deformity of the spine, club foot, polydactyly, syndactyly, epiphyseal stippling and low cholesterol (2.29 mmol/l). Spectrophotometric analysis revealed the presence of abnormal pattern of cholesterol precursors in blood. The increased level of 8-dehydrocholesterol (42.2 micromol/l, controls < 1) and 7-dehydrocholesterol (25.5 micromol/l, controls < 1) recognised with GC/MS suggested an endogenous defect of cholesterol biosynthesis. The diagnosis of X-linked dominant chondrodysplasia punctata (CDPX2) was confirmed by the molecular analysis. Sequencing of the EBP gene encoding for 3beta-hydroxysteroid-delta8,delta7-isomerase revealed the presence of "de novo" heterozygous mutation c.327C>T (p.Arg110Stop). High cholesterol diet normalized cholesterol level (3.28 mmol/l) but it had no influence on the unfavourable prognosis of the disease. Low level of cholesterol with abnormal sterol profile in a child with congenital development anomalies represent an important laboratory marker suggesting an inherited defect of cholesterol biosynthesis. PMID- 18399065 TI - Multidisciplinary orofacial therapy after suicidal treatment: casuistical report. AB - Cutting wounds and bruises in the area of neck are very specific. The overall rehabilitation of these conditions is difficult and often needs to be combined with special and customised orofacial prostheses. This case-study reports special multidisciplinary treatment. The surgical diagnosis was: a cutting wound on a neck caused by a chain saw and disruption of oropharynx. Due to contusion of the tongue root and contusion of both hypoglossal nerves, a complete paralysis of tongue occurred. The tongue was immobile and the patient has problems with swallowing fluids. A special palatal prosthesis was made to reduce the space between the tongue and the palate. The fluoroscopically monitored swallowing action helps to receive optimal patient's rehabilitation. PMID- 18399066 TI - Dynamic algorithm for inferring qualitative models of Gene Regulatory Networks. AB - We introduce a novel algorithm, DFL (Discrete Function Learning), for reconstructing qualitative models of Gene Regulatory Networks (GRNs) from gene expression data in this paper. We analyse its complexity of O(k x N x n2) on the average and its data requirements. The experiments of synthetic Boolean networks show that the DFL algorithm is more efficient than current algorithms without loss of prediction performances. The results of yeast cell cycle gene expression data show that the DFL algorithm can identify biologically significant models with reasonable accuracy, sensitivity and high precision with respect to the literature evidences. PMID- 18399067 TI - Improving domain-based protein interaction prediction using biologically significant negative datasets. AB - We propose a domain-based classification method to predict protein-protein interactions using probabilities of putative interacting domain pairs derived from both experimentally-determined interacting protein pairs and carefully chosen non-interacting protein pairs. Multi-species comparative results for protein interaction prediction show that such careful generation of biologically meaningful negative training data can improve classification performance. PMID- 18399068 TI - Spectral similarity for analysis of DNA microarray time-series data. AB - This paper proposes a new similarity measurement for comparison and analysis of DNA microarray time-series data. In this method, a gene expression time series is decomposed into frequency components and the correlation between the data from a pair of genes is measured in the frequency domain. The method effectively solves the phase delay problem and provides a more accurate metric for microarray time series classification. PMID- 18399069 TI - Transitive closure and metric inequality of weighted graphs: detecting protein interaction modules using cliques. AB - We study transitivity properties of edge weights in complex networks. We show that enforcing transitivity leads to a transitivity inequality which is equivalent to ultra-metric inequality. This can be used to define transitive closure on weighted undirected graphs, which can be computed using a modified Floyd-Warshall algorithm. These new concepts are extended to dissimilarity graphs and triangle inequalities. From this, we extend the clique concept from unweighted graph to weighted graph. We outline several applications and present results of detecting protein functional modules in a protein interaction network. PMID- 18399070 TI - BAG: a graph theoretic sequence clustering algorithm. AB - In this paper, we first discuss issues in clustering biological sequences with graph properties, which inspired the design of our sequence clustering algorithm BAG. BAG recursively utilises several graph properties: biconnectedness, articulation points, pquasi-completeness, and domain knowledge specific to biological sequence clustering. To reduce the fragmentation issue, we have developed a new metric called cluster utility to guide cluster splitting. Clusters are then merged back with less stringent constraints. Experiments with the entire COG database and other sequence databases show that BAG can cluster a large number of sequences accurately while keeping the number of fragmented clusters significantly low. PMID- 18399071 TI - An efficient motif discovery algorithm with unknown motif length and number of binding sites. AB - Most motif discovery algorithms from DNA sequences require the motif's length as input. Styczynski et al. introduced the Extended (l,d)-Motif Problem (EMP) where the motif's length is not an input parameter. Unfortunately, their algorithm takes an unacceptably long time to run, e.g. over 3 months to discover a length 14 motif. Since the best motif may not be the longest nor have the largest number of binding sites, in this paper we further eliminate another input parameter about the minimum number of binding sites in order to provide more realistic/robust results. We also develop an efficient algorithm to solve EMP and this redefined problem. PMID- 18399072 TI - Simulation study in Probabilistic Boolean Network models for genetic regulatory networks. AB - Probabilistic Boolean Network (PBN) is widely used to model genetic regulatory networks. Evolution of the PBN is according to the transition probability matrix. Steady-state (long-run behaviour) analysis is a key aspect in studying the dynamics of genetic regulatory networks. In this paper, an efficient method to construct the sparse transition probability matrix is proposed, and the power method based on the sparse matrix-vector multiplication is applied to compute the steady-state probability distribution. Such methods provide a tool for us to study the sensitivity of the steady-state distribution to the influence of input genes, gene connections and Boolean networks. Simulation results based on a real network are given to illustrate the method and to demonstrate the steady-state analysis. PMID- 18399073 TI - A parallel edge-betweenness clustering tool for Protein-Protein Interaction networks. AB - The increasing availability of protein-protein interaction graphs (PPI) requires new efficient tools capable of extracting valuable biological knowledge from these networks. Among the wide range of clustering algorithms, Girvan and Newman's edge betweenness algorithm showed remarkable performances in discovering clustering structures in several real-world networks. Unfortunately, their algorithm suffers from high computational cost and it is impractical for inputs of the size of large PPI networks. Here we report on a novel parallel implementation of Girvan and Newman's clustering algorithm that achieves almost linear speed-up for up to 32 processors. The tool is available in the public domain from the authors' website. PMID- 18399074 TI - Prediction of Protein Secondary Structure with two-stage multi-class SVMs. AB - Bioinformatics techniques to Protein Secondary Structure (PSS) prediction mostly depend on the information available in amino acid sequences. In this paper, we propose a two-stage Multi-class Support Vector Machine (MSVM) approach, where the second MSVM predictor is introduced at the output of the first stage MSVM to capture the contextual relationship among secondary structure elements in order to minimise the generalisation error in the prediction. By using position specific scoring matrices generated by PSI-BLAST, the two-stage MSVM approach achieves Q3 accuracies of 78.0% and 76.3% on the RS126 dataset of 126 non homologous globular proteins and the CB396 dataset of 396 non-homologous proteins, respectively, which are better than the scores reported on both datasets to date. By using MSVM, the present prediction scheme significantly achieves 2-6% and 3-15% of improvement in Q3 and Sov accuracies, respectively, on the two datasets. On larger blind-test datasets from PSIPRED, CASP4 and EVA datasets, two-stage MSVM approach achieves Q3 accuracies from 77.0% to 79.5%. PMID- 18399075 TI - Granular kernel trees with parallel genetic algorithms for drug activity comparisons. AB - With the growing interests of biological data prediction and chemical data prediction, more powerful and flexible kernels need to be designed so that the prior knowledge and relationships within data can be expressed effectively in kernel functions. In this paper, Granular Kernel Trees (GKTs) are proposed and parallel Genetic Algorithms (GAs) are used to optimise the parameters of GKTs. In applications, SVMs with new kernel trees are employed for drug activity comparisons. The experimental results show that GKTs and evolutionary GKTs can achieve better performances than traditional RBF kernels in terms of prediction accuracy. PMID- 18399076 TI - Exploring alternative knowledge representations for protein secondary-structure prediction. AB - Methods for 3-class secondary-structure prediction are thought to be reaching the highest achievable accuracy. Their accuracy on beta-sheet residue class is considerably lower than for the other two classes. We analysed the relevance of 315 individual input attributes for a predictor with the usual framework of using sequence-profile based data with an input window of fixed size. We propose two alternative knowledge representations with significantly smaller sets of input attributes. We also investigated the possibility of exploiting the prediction of connected pairs of beta-sheet residues and the prediction of residue contact maps for the improvement of accuracy of secondary-structure prediction. PMID- 18399077 TI - Bush Administration Medicaid budget cuts set off a firestorm. PMID- 18399078 TI - State Medicaid programs responding to the DRA changes to long-term care. PMID- 18399079 TI - Medicaid at the crossroad in Ohio. PMID- 18399080 TI - Making the case for home health care. PMID- 18399081 TI - The experience of selected states with home health care under Medicaid. PMID- 18399082 TI - ADEA--fit at 40. PMID- 18399083 TI - Unbundling medical supplies: a disease management initiative? PMID- 18399085 TI - Youman feelings. PMID- 18399084 TI - States look to telehomecare to cut Medicaid costs, improve service. PMID- 18399086 TI - Considerations for benchmarking. PMID- 18399087 TI - Wow! What's happening with patient satisfaction? PMID- 18399088 TI - The four big barriers to success in private duty with your certified agency. PMID- 18399089 TI - Sales and marketing: the toughest job I never knew I'd love. PMID- 18399090 TI - Marketing for more Medicaid hours? Try the Internet. PMID- 18399091 TI - CMS proposes regulatory cut of $2.9 billion in hospice payments: President's budget already calls for $5.14 billion cut. PMID- 18399092 TI - Miracles without fairytale endings. PMID- 18399093 TI - NAHC names protecting homebound Medicare patients as top priority. PMID- 18399094 TI - Combined radical retropubic prostatectomy and abdominoperineal excision of the rectum for locally invasive rectal cancer as a less invasive surgery: report of a case. AB - The optimal therapy for carcinoma of the rectum with invasion of the prostate gland has not been established. For a patient who has rectal carcinoma invading into the prostate and seminal vesicles and not invading into any other pelvic viscera, we performed combined radical retropubic prostatectomy and abdominoperineal excision of the rectum with reconstruction of the urinary tract by anastomosis of the ureter to the bladder. En bloc excision yielded negative surgical margins. After the operation, the patient had an infection of the abdominal wound and leakage of the anastomosis of the urethra to the bladder. These complications were treated conservatively and improved without becoming critical. The patient now has satisfactory postoperative function of voiding. This technique obviates the need for urinary diversion or urinary reconstruction such as the neobladder in the case of total pelvic exenteration. We consider this procedure is of benefit for improving the quality of life of patients with rectal cancer invading into the prostate. PMID- 18399095 TI - Patent omphalomesenteric duct and exomphalos minor: a rare and interesting association. AB - A completely patent omphalomesenteric duct was seen in association with two cases of exomphalos minor. Both the patients also had limb anomalies, and one patient had an ileal duplication and colonic atresia. Both babies underwent emergency surgery to correct the anomalies, and one survived. There have been few reports of this combination of anomalies, and there is increasing appreciation that these lesions occur together more often than by chance. PMID- 18399096 TI - Could fetal bovine serum protect the small bowel from ischemia and reperfusion injury by using a systemic perfusion. AB - The luminal perfusion of fetal bovine serum (FBS) has been reported to protect the small bowel from ischemia/reperfusion injury. We studied the effect of intravascular FBS infusion in a rat model of intestinal ischemia and reperfusion. Thirty male Lewis rats received an FBS (FBS group) or saline [control (CON) group] infusion through the vena cava. Fourteen days later (day 0), all rats underwent 2-hours total vascular occlusion in 15 cm of the intestine. Rats were killed on days 0, 3, and 6 to determine the intestinal morphology and DNA and protein content. After FBS perfusion, DNA and protein levels showed no significant increase. In both groups, the mucosa and weight of the intestine changed, but not significantly. The lung, spleen, and liver were congested. Mucosal morphology of the small intestine and DNA and protein content do not significantly change with FBS infusion. Reasons might be an insufficient dose of FBS or the short duration of intestinal ischemia. Systemically perfused FBS may also act as an antigen in some organs. The role of FBS in sysTemic perfusion requires further study. PMID- 18399097 TI - Anastomotic suture line recurrence of early rectal carcinomas that had been resected by laparoscopic surgery. AB - We report herein a case of a 64-year-old woman found to have anastomotic suture line recurrence of an early rectal carcinoma. The patient had undergone laparoscopy-assisted low anterior rectal resection for an early rectal carcinoma 2 years before the anastomotic site recurrence. A follow-up colonoscopy revealed an elevated lesion on the anastomotic suture line. The diagnosis of adenocarcinoma was confirmed by biopsy. The patient underwent a resection of the remnant rectum. Histological examination of the resected specimen showed that the anastomotic site recurrence might have been caused by intraluminal implantation from the primary rectal cancer. We speculate that intraluminal implantation might be caused by insufficient intraoperative rectal irrigation because of limited access often encountered in laparoscopic surgery. We propose that it is necessary to devise a method with which to perform sufficient intraoperative rectal irrigation in laparoscopic surgery for rectal carcinoma. PMID- 18399098 TI - Modified anterior compartment resection of the thigh. AB - For large soft tissue sarcomas of the anterior thigh, an anterior compartment resection is often performed. This may leave the patient with the inability to extend the knee. In our practice, we find that it is usually possible to preserve one of the heads of the quadriceps, usually the vastus medialis, with intact innervation, and thus preserve significant extension function of the knee joint, while the requirements for a radical oncologic procedure are being met. The modified anterior compartment resection preserves one or more heads of the quadriceps and requires a thorough knowledge of the course and distribution of the branches of the femoral nerve from the level of the inguinal ligament to their termination at the individual heads of the quadriceps femoris. PMID- 18399099 TI - Clinicopathological analysis of a case with mesenteric solitary Castleman's disease: diagnostic value of radiological findings. AB - We report a case with rare solitary mesenteric Castleman's disease (CD). A 45 year-old woman complaining of nausea was presented. A round-shaped, smooth margin, and hypoechoic mass was seen on screening abdominal ultrasonography. Computed tomography showed a markedly enhanced tumor anterior to the left iliopsoas muscle. Selective jejunal arteriography revealed an extreme hypervascularity without vascular invasion. These results retrospectively seem to differ from those of malignant lymphoma or sarcoma. The tumor was surgically resected, and hyaline-vascular type CD was pathologically diagnosed. We postulate that these radiological findings might suggest hyaline-vascular type CD as one of the differential diagnoses in similar cases, although more clinical data should be evaluated. PMID- 18399100 TI - Gene expression profile analysis of the peripheral blood mononuclear cells from tolerant living-donor liver transplant recipients. AB - Induction of transplant tolerance is a clinically desirable goal. To provide unbiased insight into transplant tolerance, we analyzed gene expression profiling in peripheral blood mononuclear cells from recipients of living-donor liver transplants (LDLTs) who had retained an immune tolerance with a well-functioning graft for several years using cDNA microarray. The comparative analyses with nontransplanted normal healthy volunteers showed that the majority of reliable detected genes were similar, and 5.6% of the genes in the tested genome (of which 627 up-regulated and 90 down-regulated) were significantly regulated and specific to tolerant LDLT recipients, indicating a significant genetic feature for inducing and maintaining immune tolerance. Moreover, the expression of several selected genes was confirmed by semiquantitative reverse transcriptase-polymerase chain reaction, which correlated to microarray data. Our data indicated that cDNA microarray technology was useful for this application and provided many informative insights into transplant tolerance mechanism. PMID- 18399101 TI - Novel mutations of the HOXD13 gene in hand and foot malformations. AB - Homeobox genes encode a set of transcription factors of fundamental importance for body patterning during embryogenesis. Hoxa9-a13 and Hoxd9-d13 play an especially important part in vertebrate limb development. Synpolydactyly (SPD) is characterized by various malformations of the limbs. The expansion of the polyalanine tract in 1OXD13 is one of its major causes. Recently, there have been many analysis studies of HOXD13 in patients with SPD and limb malformations. We analyzed HOXD13 in 100 patients with limb malformations, which affects the limbs in the distal parts of the metacarpal and/or metatarsal bones. Seven mutations in the coding region and two mutations in the 5'-untranslated region were identified. All were novel mutations. In this study, the mutations were located upstream in the homeobox. Thus, translation of the homeobox was affected by upstream mutations. Consequently, this suggested the possibility that abnormalities in the hands and feet could be caused by novel HOXD13 gene mutations. PMID- 18399102 TI - Massive mesenteric ischemia resulting from a giant strangulated umbilical hernia. AB - Hernia repair is the most commonly practiced operation in the departments of surgery in developed countries. Huge abdominal hernias are uncommon in western civilization. We present a rare case of a 73-year-old woman with a diagnosis at admission of intestinal obstruction caused by a giant strangulated umbilical hernia. At the clinical and radiological examination, the patient showed an enormous strangulated umbilical hernia with acute abdomen, atrial fibrillation, and pulmonary subedema. Emergency laparotomy showed a huge peritoneal umbilical sac containing massive mesenteric ischemia starting from 40 cm after the Treitz ligament and extended to the right colonic flexure. A near-total resection of the small bowel, a right colectomy with double terminal stomas, and a direct hernia repair without prosthetic mesh were performed. Twenty days after the operation, the patient was discharged and begun domiciliary total parenteral nutrition, and 24 months after surgical treatment she is still alive. PMID- 18399103 TI - Tumoral calcinosis of the shoulder. AB - Tumoral calcinosis is a rare clinical and histopathologic syndrome that causes the formation of calcium salt crystal deposits in periarticular soft tissue. This soft tissue calcification typically behaves as a large-sized tumor. However, the mechanism is unknown for the massive periarticular calcification caused by tumoral calcinosis. In addition, tumor calcinosis occurs infrequently in patients who have been on long-term hemodialysis and have end-stage renal disease (ESRD). This report describes a 40-year-old male uremia patient on long-term hemodialysis. This patient developed a huge tumor mass and pain over the left shoulder and was diagnosed with tumoral calcinosis. Although common treatments include management of metabolic levels such as calcium and phosphate, in some cases, surgical intervention may be needed. In this patient case, complete surgical excision was done successfully with an excellent outcome at the 2-year follow-up. PMID- 18399104 TI - A fast and safe living donor "finger assisted" nephrectomy technique. PMID- 18399105 TI - Napoleon Bonaparte really was murdered. The weapon: rat poison. PMID- 18399106 TI - [Applying clinical pathway to promote the reform of payment mode in medical insurance system]. PMID- 18399107 TI - [Mid-, and long-term effects of video-assisted thoracoscopic and transsternal thymectomy in treatment of non-thymomatous myasthenia gravis: analysis of 204 cases]. AB - OBJECTIVE: To evaluate the mid-, and long-term effects of video-assisted thoracoscopic thymectomy and transsternal thymectomy in treatment of myasthenia gravis (MG) and to identify the prognostic factors for thymectomy success. METHODS: 161 patients with non-thymomatous MS, 84 males and 120 females, aged 33, underwent transsternal thymectomy and were followed up for 5 years; and 43 patients with non-thymomatous MS A retrospective, 1 male and 25 females, 21 being aged >40 during operation, underwent thoracoscopic thymectomy and were followed up for 3 years. RESULTS: The mean operating time of the thoracoscopic group was (132 +/- 32) min minutes, significantly longer than that of the transsternal thymectomy group [(96 +/- 18) min, P = 0.000]. Four patients in the transsternal thymectomy group and 41 in the transsternal thymectomy group developed myasthenic crises (P = 0.023). The complete stable remission (CSR) rates 1, 2, and 3 years after operation of the thoracoscopic thymectomy group were 34.9%, 41.9%, and 46.5% respectively; and CSR rates 1, 2, 3, and 5 years after operation of the transsternal thymectomy group were 26.7%, 31.7%, 35.4%, and 40.4% respectively, without significant differences between these 2 groups. CONCLUSION: Both thoracoscopic and transsternal approaches to thymectomy in patients with MG are effective in terms of remission. The authors advocate adopting the thoracoscopic approach early. PMID- 18399108 TI - [Reducing oxidative DNA damage by adding antioxidants in human semen samples undergoing cryopreservation procedure]. AB - OBJECTIVE: To evaluate the protective effects of oxidative DNA damage by adding antioxidants: ascorbate, catalase (CAT), and superoxide dismutase (SOD) in human semen samples undergoing cryopreservation procedure. METHODS: Semen sample form 30 fertile men were mixed with modified cryoprotectant and divided into six groups according to the category and concentration of antioxidants: ascorbate 300 micromol/L, ascorbate 600 micromol/L, CAT 200 U/ml, CAT 400 U/ml, SOD 200 U/ml, and SOD 400 U/ml. Comet assay was conducted to measure the percentage of comet cells, and the nuclear DNA damaged parameters: tail DNA percentage (TD%) and Olive tail moment (OTM). Flow cytometry was used to detect the reactive oxidative species (ROS). The motility (a + b grade), viable recovery rate, nuclear DNA integrity and reactive oxidative species (ROS) of all groups were analyzed before and/or after freeze-thawing. RESULTS: (After cryopreservation, compared with the control group, the a + b grade sperm rates of the ascorbate 300 micromol/L, CAT 200 U, and CAT 400 U groups were all higher than that of the control group (all P < 0.05), however, the levels of reactive oxygen species (ROS) of the ascorbate 300 micromol/L, CAT 200 U, and CAT 400 U groups were 30 +/- 13, 30 +/- 11, and 30 +/- 11 respectively, all significantly lower than that of the control group (37 +/- 17 , all P < 0.05). The viable recovery rates of the ascorbate 300 micromol/L , CAT 200 U, and CAT 400 U groups were 67% +/- 14%, 68% +/- 14%, and 69% -/+ 15% respectively, all significantly higher than that of the control group (59% +/- 10%, all P < 0.05). (2) The TD% levels of the ascorbate 300 micromol/L, CAT 200 U, and CAT 400 U groups were 41% +/- 4%, 40% +/- 7%, 40% +/- 6%, all similar to that of the raw semen (all P > 0.05), but significantly lower than that of the control group (46% +/- 6%, all P < 0.01). The OTM levels of the ascorbate 300 micromol/ L, CAT 200 U, and CAT 400 U groups were 7.7 +/- 1.2, 7.5 +/- 1.6, and 7.8 +/- 1.9, all similar to that of the raw semen (all P > 0.05), but significantly lower than that of the control group (10.1 +/- 3.1, all P < 0.01) too. The TD% and OTM levels of the other groups were all significantly higher than that of the raw semen (all P < 0.01), but not significantly different from those of the control group (all P > 0.05). (3) ROS was significantly negatively correlated with the motility in all groups (P < 0.05 or P < 0.01). Apart from the ascorbate 600 micromol/L group, the TD% and OTM of the other groups were all significantly positively correlated with the ROS (P < 0.05 or P < 0.01). CONCLUSION: Supplementation of ascorbate or CAT reduces the level of ROS that induces sperm nuclear DNA damage, and improves the human sperm quality in the process of freeze-thawing. PMID- 18399109 TI - [Clinical study and prenatal diagnosis of fetus with shortened long bones]. AB - OBJECTIVE: To investigate the manifestations and prenatal diagnosis of the fetus with shortened long bones. METHODS: Ultrasonography was conducted on 57 fetuses with shortened long bones and 62 normal fetuses. Percutaneous umbilical blood sampling was performed on the 57 cases to undergo examination of karyotype and mutation in exon 10 of fibroblast growth factor receptor (FGFR)3 gene. Fetal radiography and autopsy were performed in 30 abnormal induced fetuses. RESULTS: Among the 57 fetuses of the shortened long bones group 21 (37%) were diagnosed as with specific skeletal dysplasia postnatally, abnormal chromosome were found in 8 (14%), 4 (7%) were with fetal growth restriction, and 24 (42%) were with unknown etiology. Prenatal ultrasonography showed that in the fetuses with skeletal dysplasia the value of standard deviation score (SD) for femur length and/or humerus length fell to -4 SD of the corresponding pregnancy week, and increase of biparietal diameter was often found; in the fetuses with abnormal chromosome and fetal growth restriction (FGR) the degree of bone shortening was between -2 SD to -4 SD; and in the fetuses with lethal skeletal dysplasia the long bones were shortened at the degree more than -8 SD. When the ratio of femur length to abdominal circumference was less than 0.16, the sensitivity and specificity for predicting narrow thorax were 100% and 85.7% respectively. CONCLUSION: The causes of fetus with shortened long bones include abnormal chromosome, skeletal dysplasia, and FGR. Femur length and/or humerus length falls below -4SD is almost an indication for skeletal abnormality. When the SD is between -2SD and -4SD, chromosome analysis is necessary and regular ultrasonography is recommended. When the femur length and/or humerus length is below -8SD and femur length/abdominal circumference (FL/AC) is less than 0.16, lethal skeletal dysplasia is suspected. PMID- 18399110 TI - [Executive function impairment in patients with temporal lobe epilepsy: neuropsychological and diffusion-tensor imaging study]. AB - OBJECTIVE: To explore the cognitive changes and the relation with the parameters of DTI in the brain areas in patients with temporal lobe epilepsy by neuropsychological test and DTI study. METHODS: Thirty-two patients with temporal lobe epilepsy, 11 males and 8 females, aged 32 +/- 12, with the education year of 8.6 +/- 1.2, and 42 sex, age, and education level-matched healthy controls underwent and a comprehensive test battery test including verbal fluency (VI), digit span (DSp), digit symbol (DSy), Stroop color-word test, trail making test, and so as to evaluate the intelligence level. Nineteen patients and twenty-one control subjects underwent diffusion tensor imaging (DTI) of the head. The correlation between the results of DTI and cognitive function tests of the patients was analyzed. RESULTS: VI test showed that the number of word spoken in 1 min by the healthy control group was (16 +/- 4), significantly more than that of the patient group [(11 +/- 4), P = 0.00]. The DSp score of the control group was (14.6 +/- 3.1) points, significantly more than that of the patient group [(11.1 +/- 2.8), P = 0.00]. The number of DSy filled by the patient group was (47 +/- 17), significantly less than that of the control group [(60 +/- 16), P = 0.00]. The time needed to make trail of the patients was (56 +/- 20) s, significantly longer than that of the control group [(37 +/- 11) s, P = 0.01]. The Stroop reaction time of the patient group was (6.1 +/- 4.5) s, significantly longer than that of the control group [(30 +/- 5) s, P = 0.00]. The Stroop error number of the patient group was (6.1 +/- 4.5), significantly more than that of the control group [(1.4 +/- 1.2), P = 0.00]. The fractional anisotropy (FA) values of the posterior limb of left internal capsule and of the bilateral thalami of the patient group were significantly lower than those of the control group (P = 0.01 or P = 0.00). VI was negatively correlated with the mean diffusivity (MD) in left caudamen (r = -0.56, P = 0.04), right putamen (r = 0.58, P = 0.04), left putamen (r = -0.58, P = 0.04), and right thalamus (r = 0.64, P = 0.02), and was positively correlated with the FA in white matter of left frontal lobe (r = 0.43, P = 0.04)and left occipital lobe (r = 0.47, P = 0.02). DSp was negatively correlated with the MD in splenium of corpus callosum (r = 0.58, P = 0.04) and left putamen (r = -0.59, P = 0.04). TMT was positively correlated with the MD in the posterior limb of right internal capsule (r = 0.65, P = 0.02) and posterior limb of left internal capsule (r = 0.59, P = 0.03). CONCLUSION: The prefrontal cortex is a vital component of the circuitry subserving executive function. But the corresponding damaged area in patients with impaired executive function was not limited to frontal lobe,more areas beyond frontal lobe may be involved in executive function. PMID- 18399111 TI - [Clinical and electrophysiological characteristics of myasthenia gravis with thymoma: analysis of 72 patients]. AB - OBJECTIVE: To investigate the clinical and electrophysiological characteristics of myasthenia gravis with thymoma (MGT). METHODS: The clinical and repetitive nerve stimulation (RNS) data of 72 patients with MGT, 36 males and 36 females, and 63 patients with myasthenia gravis without thymoma (NMGT) were analyzed retrospectively. RESULTS: The onset age of 52 of the 72 MGT patients (72.2%) was 35-59. The average onset age of the MGT patients was 39.5, older than that of the NMGT patients (29.4 yrs), and the course of the MGT patients was 13.1 months, shorter than that of the NMGT patients (29.1 months)The attack rate of the males was not significantly different than that of the females. The proportion of generalized presentation type among the MGT patients was 72.8%, higher than that among the NMGT patients (66.7%), particularly concerning the respiratory muscle (20.8% vs. 9.5%). The average time interval between the onset of symptoms and respiratory or bulbar muscle involvement of the NMGT patients were 12 and 7 months respectively, both remarkably shorter than those of the NMGT patients (26.4 and 11.6 months respectively). The abnormal RNS test yield rate of the MGT patients was 86.9%, higher than, however, not significantly that of the NMGT patients (75.0%). 97.2% of the thymoma and mediastinal abnormality could be detected through computed tomography (CT) scan on thymus. The positive rate of AchRab titer of the MGT patients was 50.0%, not significantly different from that of the NMGT patients (52.4%). CONCLUSION: MGT is common in middle-aged people, both male and female. The Course is quickly progressive and generalized skeletal muscles, particularly respiratory muscles, are more involved in the MGT patients than in the NMGT patients. RNS study is a sensitive tool to identify MGT. AchRab may not be the specific antibody for recognizing MGT. CT scan is useful in differentiating thymoma preoperatively. PMID- 18399112 TI - [Measurement of alveolar-arterial partial pressure of oxygen difference in patients with chronic obstructive pulmonary disease both at rest and during maximal exercise]. AB - OBJECTIVE: To investigate the change of alveolar-arterial partial pressure of oxygen (PO2) difference [P (A-a) O2)] at rest and during exercise in patients with chronic obstructive pulmonary disease (COPD). METHODS: Cardiopulmonary exercise testing was performed in 47 COPD male patients aged (66 +/- 8) at stable stage to measure the oxygen uptake (VO2) and carbon dioxide output (VCO2) continuously in a breath-by-breath mode. Arterial blood samples were drawn both at rest and during maximal exercise. P(A-a) O2 is computed by the equation: PAO2 PaO2. RESULTS: The PaO2 level during exercise was (89 +/- 14) mm Hg, a little bit, however, not significantly, lower than that at rest [(92 +/- 9) mm Hg, P = 0.506]. The PaCO2 during exercise was 43 +/- 3 mm Hg, significantly higher than that at rest (41 +/- 4 mm Hg, P = 0.003). The patients were divided into two groups according to lung diffusing capacity for carbon monoxide (DLco). There was a significant increase in P(A-a) O2 from (16 +/- 8) mm Hg at rest to (42 +/- 9) mm Hg during maximal exercise in the DLco < 80% group (P = 0.005); however, in the DLco >80% group the P(A-a) O2 level during maximal exercise was (26 +/- 6) mm Hg, not significantly different from that at rest [(20 +/- 6) mm Hg, P = 0.106]. The P(A-a)O2 level of the DLco <80% group during maximal exercise was (42 +/- 9) mm Hg, significantly higher than that at rest [(16 +/- 8) mm Hg, P = 0.005]. The P(A-a)O2 was significantly negatively correlated with the forced vital capacity (r= -0.581, P = 0.037) and DLco (r = -0.671, P = 0.012). CONCLUSION: The increase in P (A-a) O2 during exercise in the COPD patients is mainly due to the limited diffusing capacity of the lung. PMID- 18399113 TI - [Surgical treatment of medial collateral ligament laxity complicated by old cruciate ligament injury of the knee]. AB - OBJECTIVE: To investigate the therapeutic effect of transposition of semitendinosus tendon combined with femur insertion of medial collateral ligament (MCL) and reconstruction of anterior cruciate ligament (ACL) with autograft bone patellar tendon-bone in treatment of MCL laxity complicated by ACL injury of the knee. METHODS: Twenty patients (20 knees), 14 male and 6 female, aged 32.6 (16 64), 1 case at grade II and 19 cases at grade III according to X ray classification, all cases at grade III by MRI evaluation, and with preoperative Lysholm knee joint score of 48.9 (35-65), underwent transposition of semitendinosus tendon combined with femur insertion of medial collateral ligament (MCL) and reconstruction of anterior cruciate ligament (ACL) with autograft bone patellar tendon-bone. Followed up was conducted for 19.2 (6-59) months. RESULTS: The postoperative Lysholm score was 83.9 (50-96). Eighteen of the 20 patients obtained an excellent result with good function of the knee joint except one case with osteoarthritis and one case with limited range of motion of the knee (90 degree), and the good-to-excellent rate was 90%. CONCLUSION: Reconstruction with autograft of the bone-patellar tendon-bone and transposition of semitendinosus tendon combined with femur insertion of MCL proximal transfer is a reliable treatment method for MCL laxity complicated by old ACL injury of the knee. PMID- 18399114 TI - [Long-term clinical and radiological results of entirely hydroxyapatite-coated femoral components]. AB - OBJECTIVE: To evaluate the clinical and radiological results of entirely hydroxyapatite (HA)-coated femoral prosthesis. METHODS: Thirty-five patients aged 59.7 (32 -79), underwent total hip arthroplasty using HA-coated femoral prosthesis, 32 of which were followed up for 10.2 (9.7-10.9) years. Harris scoring and X-ray examination were used to evaluate the clinical and radiological results. RESULTS: The symptoms of all the patients were alleviated. One patient complained nonspecific discomfort in groin area, and 2 patients showed mild limp. One patient had to undergo pure repair of acetabulum 7 years later. No thigh pain was seen in other cases. The mean Harris score 10 year after operation was 87.70 +/- 7.73, significantly higher than that before operation (39.20 +/- 8.88, P < 0.05). No case of aseptic loosening was found. X-ray examination showed no osteolysis and radiolucent line. Subsidence of prosthesis was found in 3 cases; however, with a distance less than 2 mm. The 10-years survivorship of the entirely HA-coated prosthesis was 100%. CONCLUSION: Entirely HA-coated total hip prosthesis has an excellent clinical and radiological long term results. PMID- 18399115 TI - [Expression profile of clock genes during the process of embryonic stem cells differentiation]. AB - OBJECTIVE: To analysis the expression profile of7 clock genes during mouse embryonic stem cell (mES) differentiation. METHODS: Mouse ES cells of the line 129 were cultured and induced to differentiate into neurons by 5 stages method. The expression of 7 clock genes: BMAL1, CLOCK, CRY1, CRY2, PER1, PER2, and PER3 in the five stages were determined by RT-PCR. RESULTS: In the amplification condition, no non-specific band was found, and only bands of expected sizes were detected. Six clock genes, BMAL1, CLOCK, CRY1, CRY2, PER1 and PER3 were expressed in 129 cells at all five stages in the differentiation process, however, PER2, could only be determined in the last stage. CONCLUSION: The transcription profile of the clock genes is different during the differentiation of the ES cells towards neurons. The transcription of the gene PER2 is limited to the final stage, when postmitotic neurons and astrocytes emerged. A clock genes loop existing in the mature cells has not been established during the process of ES cell differentiation. PMID- 18399116 TI - [Autologous sinoatrial node cells heterotopic transplantation for treating bradycardia: an experiment with dogs]. AB - OBJECTIVE: To explore the feasibility of autografting sinoatrial nodal cells heterotopic transplantation to construct an ectopia pacemaker for treating bradycardia. METHODS: Sixteen healthy adult dogs were randomly divided into 2 equal groups: graft group and control group. The sinoatrial node (SAN) of the dogs in the graft group was harvested and digested into cell suspension in vitro, then injected to the autogenic right ventricular wall adjacent to heart apex. Commensurable culture medium was injected to the same position with the dogs in control group. Two week later, all dogs underwent transcatheter ablation of His bundle to create a complete heart block model and an electrophysiology study was carried out. In order to investigate the change of rhythm, isoproterenol and atropine was injected respectively to dogs of the graft group. Two weeks later the dogs were killed with their hearts taken out. Immunofluorescence histochemistry was used to investigate the survival of grafted cells and gap junction formed between grafted cells and ventricular myocytes. RESULTS The isolated cells from SAN retained active and beating. After ablation, the heart rate of the dogs of the graft group was (91 +/- 14) bpm, significantly higher than that of the control group, [(49 +/- 11) bpm, t = 6. 672, P < 0.01], and electrocardiography showed that these ventricular rhythms originated from the cell transplant sites. After the injection of isoproterenol the ventricular rate of the graft group was (118 +/- 15) bpm, significantly higher than that before the injection, (95 +/- 11) bpm, t = 3.491, P < 0.01), however, after the injection of atropine, the heart rate of the graft group was (101 +/- 17) bpm, not significantly different from that before the injection, [(95 +/- 11) bpm, t = 0.838, P > 0.05]. Immunofluorescence staining showed that the grafted autografting sinoatrial nodal cells all survived and that there was connexin-43 expression among the cells. CONCLUSION: Transplantation of autologous SAN cells into the right ventricular wall can boost the ventricular rhythm which is sensitive to isoproterenol but not to atropine. Grafted SAN cells can form gap junctions with adjacent myocytes. PMID- 18399117 TI - [In vivo MR imaging tracking of supermagnetic iron-oxide nanoparticle-labeled bone marrow mesenchymal stem cells injected into intra-articular space of knee joints: experiment with rabbit]. AB - OBJECTIVE: To evaluate the feasibility of in vivo magnetic resonance imaging (MRI) with 1.5T system tracking of the survival, migration and differentiation of magnetically labeled seed cells-bone marrow-derived mesenchymal stem cells (MSCs) injected into the articular cavity. METHODS: Rabbit MSCs were isolated, purified, expanded, and then coincubated in vitro with supermagnetic iron oxide particles (SPIO) and 5-bromo-2-deoxyuridine (BrdU). Prussian blue staining and transmission electron microscopy were performed to observe the intracellular iron. Some labeled MSCs were subjected to chondrogenic differentiation and the phenotype was examined to assess their chondrogenic differentiation capacity. MSCs colabeled with SPIO nanoparticles and (BrdU were suspended in chitosan and glycerophosphate (C-GP) gel. Eighteen rabbits underwent damage to the femoral trochlea to create cartilage defect models, and randomly divided into 3 groups 1 week later: Group A (n=6) undergoing injection of the MSC suspension in C-GP gel into the intra articular space of knee joints, Group B (n=6), injected with un-labeled MSC suspension in C-GP gel, and Group C (n=6), without injection. MRI of the knee was performed 1, 4, 8, and 12 weeks after the injection respectively on a certain numbers of rabbits. and then the rabbits were killed with their knee joints taken out to undergo immunohistochemistry. The MR imaging findings were compared with the histological findings. RESULTS: Prussian blue staining and transmission electron microscopy showed intracytoplasmic nanoparticles in the SPIO-labeled cells. Safranin-O staining showed deposition of proteoglycan and type II collagen outside both the labeled and unlabeled MSCs, showing chondrogenesis. GRE T2 weighted MR image showed marked hypointense signal void areas, representing the implanted MSCs, in the intra-articular space after the MSC injection in Group A that persisted for 12 weeks at least; 2 week after the MSC injection hypointense signal could be seen in the defect, which peaked in the signal intensity about 4 weeks later, and then gradually decreased in the signal intensity; and 12 weeks after the injection no recognizable hypointense signal in the defect was detected. Immunohistochemical staining demonstrated the presence of Prussian blue positive cells and BrdU-positive cells in the tissue sections in the areas corresponding well to the signal intensity loss regions in the MRI images. Group B and Group C showed no signal intensity loss in the intra-articular spaces by GRE T2-weighted MR imaging. Histological observation showed that the defects were repaired with fibrocartilage in Groups A and B, and with fiber tissue in Group C. CONCLUSION: Labeled with SPIO, the MSCs remains their ability of chondrogenic differentiation. It is feasible to track the fate and dynamic redistribution of magnetically labeled MSCs, the seed cells, injected into the articular cavity by 1.5T MRI, an efficient noninvasive technique. PMID- 18399118 TI - [Protein and mRNA expression of osteopontin in lung cancer and clinical significance thereof]. AB - OBJECTIVE: To investigate the protein and mRNA expression of osteopontin (OPN) in the lung cancer tissue and explore the roles thereof in the development and progression of lung cancer. METHODS: Immunohistochemistry and in situ hybridization were used to detect the protein and mRNA expression of OPN in 57 specimens of lung cancer tissue, 30 specimens of inflammatory pseudotumors and 20 specimens of pulmonary bulla, all obtained during operation. RESULTS: The OPN protein expression rate of the lung cancer tissue was 57.9% (33/57) , significantly higher than that of the inflammatory pseudotumor (16.7%, 5/30, chi2 = 13.581, P = 0.000). The OPN mRNA expression rate in the lung cancer tissue was 71.9% (41/57), significantly higher than that of the inflammatory pseudotumor (30.0%, 9/30, chi2 = 14. 138, P = 0.000). All the 20 specimens of pulmonary bullae were negative in the expression of OPN, both at the protein and mRNA levels. The OPN protein and mRNA expression rates of the lung cancer tissues with lymph node metastasis were 71.1% (27/38) and 86.8% (33/38) respectively, both significantly higher than those of the lung cancer tissue without lymph node metastasis [31.6% (6/19) and 42.1% (8/19) respectively, chi2 = 6.558, P = 0.010, and chi2 = 10.438, P = 0.001]. The OPN protein and mRNA expression rates of the non-small lung cancer tissues were 68.1% (32/47) and 78.7% (37/47) respectively, both significantly higher than those of the small lung cancer tissues [10% (1/10) and 25% (4/10) respectively, chi2 = 11.412, P = 0.001, and chi2 = 6.124, P = 0.013]. The OPN protein expression was positively correlated with the OPN mRNA expression in the lung cancer tissues (r = 0.623, P = 0.001). The 57 patients with lung cancer after surgery were followed up for 28 (24-40) months, 8 of the 32 patients with both positive OPN protein and mRNA expression had recurrence and 13 patients had a distant metastasis, while only 1 of the 15 cases negative in both OPN protein and mRNA expression showed recurrence (chi2 = 14.258, P = 0.000). 12 patients died in the both positive expression group but no patient died in the both negative expression group (chi2 = 7.554, P = 0.006). CONCLUSION: Over-expressions of OPN protein and OPN mRNA are found in lung cancer tissues and their expressions are correlated to the prognosis and metastasis of lung cancer. PMID- 18399119 TI - [Protective effects of hydroxyethyl starch (130/0.4) against ischemia reperfusion injury of kidney: experiment with rats]. AB - OBJECTIVE: To explore the protective effects of hydroxyethyl starch (HES)(130/0.4) against ischemia reperfusion injury of kidney and its impact on the expression of intercellular adhesion molecule-1 (ICAM-1). METHODS: Forty SD rats underwent resection of the right kidney and clamping of the left renal artery for 60 min to establish renal ischemia reperfusion model, and then were randomly divided into 4 equal groups: Group S, undergoing infusion of normal saline 20 mg/kg 20 min before the reopening of the renal artery, Group W1, undergoing infusion of HES 5 mg/kg 20 min before the reopening of the renal artery, Group W2, undergoing infusion of HES 10 mg/kg, and Group W3, undergoing infusion of normal saline 20 mg/kg. Ten rats underwent sham operation and were used as controls (Group C). 24 h after reperfusion arterial blood samples were collected to detect 6 the levels of serum creatinine (Cr) and blood urea nitrogen (BUN), and then the rats were killed with their kidneys taken out to undergo pathological examination and calculation of Paller's score. Immunohistochemistry was used to detect the expression of intercellular adhesion molecule-1 (ICAM-1). RESULTS: The levels of BUN and Cr of Group S were 152 +/- 22 and 3.17 +/- 1.00 mmol/L respectively, both significantly higher than those of Group C, and the levels of BUN and Cr of Groups W2 and W3 were 99 +/- 23 and 1.82 +/- 0.86 mmol/L and 92 +/- 28 and 1.57 +/- 0.70 mmol/L respectively, all significantly lower than those of Group S (P < 0. 05 or P < 0.01). The Paller's scores of W2 and W3 were 33.6 +/- 16.6 and 29.2 +/- 12.3 respectively,both significantly lower that of Group S (43.2 +/- 15. 8, both P < 0.05). The absorption levels of ICAM-1 of Groups W2 and W3 were 182 +/- 22 and 161 +/- 25 respectively, both significantly lower than that of Group S (212 +/- 32, P < 0.01). CONCLUSION: Infusion of 10-20 ml/kg HES (130/0.4) significantly alleviates the ischemia-reperfusion injury, probably by reducing the ICAM-1 expression. PMID- 18399120 TI - [Influence of overexpressed coagulant and fibrolytic components in tumor tissues on the prognosis of non-small cell lung cancer]. AB - OBJECTIVE: To evaluate the expression of tissue factor (TF), urokinase-type plasminogen activator (uPA), and urokinase-type plasminogen activator receptor (uPAR) in non-small cell lung cancer (NSCLC) tissues and to find their roles in lymph node metastasis, vascular involvement and prognosis. METHODS: Immunohistochemistry was used to examine the expression of TF, uPA, and uPAR in the tumor tissues of 97 NSCLC patients obtained during operation and 40 samples of normal lung tissues at least 5 cm away from the tumor tissues. The correlations of expression of TF, uPA, and uPAR with the clinicopathologic parameters were analyzed by chi2 test. The survival rates were calculated by Kaplan-Meier method. RESULTS: TF, uPA, and uPAR were diffusely expressed in the carcinoma cell cytoplasm with the positive rates of 61.9%, 58.8%, and 61.9% respectively; however, they were only weakly expressed in the scattered macrophage and fibroblast cells in the normal lung tissues. TF expression was correlated with tumor angiogenesis as measured by microvessel density (P < 0.01); TF(34/47), uPA(33/47), and uPAR (39/47) expressions were all positively correlated with lymph node metastasis (P < 0.05, P < 0.05, and P < 0.01), and the uPAR expression was positively correlated with vascular involvement (P < 0.01). The agreement between TF and uPAR expression was significant (r = 0.432, P < 0.01). Co-expression of TF and uPAR was significantly correlated with lymph node metastasis and vascular involvement. Kaplan-Meier survival analysis showed that median the survival time of the patients with TF, uPAR and TF-uPAR positive tumor was shorter than that of the patients with TF, uPAR and TF-uPAR negative tumors (P < 0.01). CONCLUSION: TF promotes angiogenesis, and uPAR contributes to lymph node metastasis and vascular involvement. Co-expression of TF and uPAR may play an important role in the metastasis and prognosis of NSCLC. PMID- 18399121 TI - [Development of food contamination monitoring, strengthening exposure evaluation]. PMID- 18399122 TI - [Determination of bisphenol A and alkyl phenols in canned food with high performance liquid chromatography--fluorescence]. AB - OBJECTIVE: To establish a comprehensive analytical high performance liquid chromatography fluorescence detection (HPLC-FL) in detecting bisphenol A (BPA), nonylphenol (NP) and octylphenol (OP) in canned food sold in Beijing markets. METHODS: BPA, NP and OP was extracted with methanol and dichloroacetamide and concentrated. The samples were purified on an solid extraction cartridges. The HPLC system consisted of Waters XTerra MS C18 column, a mixture of methanol and water as mobile phase and fluorescence detector with the excitation and emission wavelength at 225 nm and 310 nm respectively. RESULTS: The method established had a linear relationship, showing the detection limit of BPA, OP and NP being 0.5, 0.1 and 0.1 microg/kg in canned vegetable and instant noodle and 1, 0.5 and 0.5 microg/kg in canned fish and meat can, respectively. The recoveries of BPA, NP and OP were 74.9%-95.1% , 76.3%-103.6% and 72.1%-109.2%. The precision was 4.98% 11.2% , 2.35%-8.88% and 5.61%-12.3%, respectively. CONCLUSION: The method is simple with high sensitivity and selectivity, suitable for the determination of NP, OP and BPA in canned food. PMID- 18399123 TI - [Study on the contamination level and intake of organotins of Chinese dietary]. AB - OBJECTIVE: To obtain the baseline data of organotins' pollution of Chinese meal in order to carry on primary danger analysis of the exposure. METHODS: The samples of the third Chinese total diet study were determined by gas chromatography pulsed flame photometric detector to estimate dietary intake of organotins. The dietary intake of organotins was estimated according to the contamination level of organotins and food consumption. RESULTS: Only several kinds of organotin were founded in several foods and no organotins was found in fruit, sugar and alcoholic beverages. Dimethyltin (DMT) were detected in some samples from Southern 1 area, the content ranged from 1.5 microg/kg to 4.1 microg/kg. Butyltin compounds existed in seafoods from Southern 1 area, the contents of tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MTB) being 0.9 microg/kg, 1.1 microg/kg, 1.4 microg/kg respectively. The lower limit and upper limit of exposure to tributyltin were from 0.003 microg x kg(-1) x d(-1) to 0.006 microg x kg(-1) x d(-1) and from 0.004 microg x kg(-1) x d(-1) to 0.019 microg x kg(-1) x d(-1) respectively. Comparing to ADI of tributyltin (WHO), the Chinese dietary intake of tributyltin only accounted for 2.5% and that of butyltin only accounted for 3.5%. To identify the contamination source of organotins in Southern 1 area, the individual samples of aquatic food from individual province were analyzed, revealing that Fujian province and Shanghai City were the main contributors of organotins pollution in this area. The belt fish and yellow croaker were typical pollution samples. Higher levels of DMT were detected in seafood samples from Shanghai. CONCLUSION: The exposure level of Chinese dietary was relative low, however the sources of organotin pollution needs further investigation. PMID- 18399124 TI - [Lead and cadmium pollution in edible fungus sold in Beijing]. AB - OBJECTIVE: To investigate the lead and cadmium pollution in edible mushrooms sold in Beijing. METHODS: 146 samples of 14 species were collected form 25 markets during the period of Mar. through May, 2007 in Beijing. The pollution of lead and cadmium were analyzed respectively according to the standard of GB/T5009. 12-2003 and GB 7096-2003. RESULTS: The content of lead and cadmium in edible mushrooms was ND--1.592 mg/kg, ND--0.550 mg/kg, respectively, both lower than the allowable content prescribed by The National Ministry of Health. CONCLUSION: The contents of lead and cadmium in the mushrooms marketed in Beijing are in safe ranges. It is worthy of mentioning the variation coefficients of heavy metal concentrations existing in edible mushrooms. PMID- 18399125 TI - [Multiplex real-time PCR detecting Salmonella, Shigella and diarrheagenic Escherichia coli]. AB - OBJECTIVE: To develop a multiplex real-time PCR for the detection of Salmonella invasion protein A gene (invA), enterotoxigenic Escherichia coli (ETEC) heat labile I enterotoxin gene (elt), and Shigella or enteroinvasive E. coli (EIEC) invasive plasmid antigen H gene (ipaH). METHODS: Under the optimized reaction conditions of the multiplex real-time PCR, invA, elt, and ipaH were determined in 10-fold series of dilution of DNA extracted from Salmonella enterica serovar Typhimurium, ETEC 44815 strain and Shigella F301 strain. The three genes were examined in 90 fecal samples from diarrhea patients using the multiplex real-time PCR. When PCR-positive samples were found, the target strains were isolated and identified. RESULTS: The detectable concentration for this multiplex real-time PCR was 10 CFU/microl for Shigella F301 strain, 10(2) CFU/microl for S. enterica serovar Typhimurium and ETEC 44815 strain, respectively. Out of 90 fecal samples from diarrhea patients, thirteen were found positive for elt gene (14.4%), and five were found positive for ipaH gene (5.6%). Three E. coli strains positive for elt gene and four E. coli strains positive for ipaH gene were isolated successfully from the PCR-positive samples mentioned above. The detection of invA, elt and ipaH genes was completed in 10 h, which included an enrichment period of 6 h. CONCLUSION: The multiplex real-time PCR assay can detect invA, elt, ipaH simultaneously in a single reaction, moreover, it can detect for virulence genes in strains of Salmonella, ETEC, and Shigella or EIEC and screen these pathogens in fecal specimens from patients with diarrhea with a high specificity. PMID- 18399126 TI - [Effects of perfluorooctane sulfonate on Glu, PKC and PKA activities in mouse brain]. AB - OBJECTIVE: To study the effects of perfluorooctane sulfonate (PFOS) on contents of glutamate and activity of protein kinase C (PKC) and A (PKA) and ultrastructure injury in the brain of male mice and to explore the mechanism of neurotoxicity and patho-alteration resulted from PFOS. METHODS: 44 male mice were randomly divided into four groups, who were respectively orally given 0, 5, 10, 20 mg/kg PFOS for 10 days. The Glu consents in the brain of the mice was measured with spectrophotometer and protein kinases activity were measured with non radioactive assay of protein kinase and the changes of cerebral cortex ultrastructure were observed. RESULTS: Contents of Glu in 10 and 20 mg/kg groups were (1.57 +/- 0.11) and (1.62 +/- 0.16) mmol/g prot respectively,which was significantly increased compared with the corresponding controlled group [(1.45 +/- 0.13) mmol/g prot] (F = 39.59, P < 0.05). PKC activity in 5, 10 and 20 mg/kg BW groups were (29.05 +/- 2.89), (33.65 +/- 3.82) and (34.20 +/- 3.16) pmol x min(-1) x (mg prot)-1 respectively, which was significantly increased compared with the corresponding control group [(24.53 +/- 2.88) pmol x min(-1) x (mg prot) 1] (F = 7.75, P < 0.05). Compared with the corresponding control group, PKA in 5, 10 and 20 mg/kg BW groups increased by (24.12 +/- 3.86)%, (34.02 +/- 3.04)% and (33.42 +/- 3.71)% with a statistical significance (F = 26.27, P < 0.01). The exposed mice had cerebral cortex ultrastructure injury of cell nucleus envelope hollow. CONCLUSION: Exposure to PFOS increases Glu contents and activity of PKC and PKA in mouse brain and induce the cerebral cortex ultrastructural injury, a possible mechanism of the neurotoxicity caused by PFOS. PMID- 18399127 TI - [Study on inhibitory effects of c9, t11-conjugated linoleic acid on migration of human gastric carcinoma cell line via cyclooxygenase-2 pathway]. AB - OBJECTIVE: To study the inhibitory effects of c9, t11-conjugated linoleic acid (c9, t11-CLA) on migration of human gastric carcinoma cell line (SGC-7901) via cyclooxygenase-2 (COX-2) pathway. METHODS: After inhibiting COX-2 activity by 100 micromol/L COX-2 inhibitor NS-398 in SGC-7901 cell, we treated SGC-7901 cells with c9, t11-CLA at a concentration of 200,100, 50, 25 micromol/L for 24 h, respectively. Using reconstituted basement membrane invasion, adhesion, chemotaxis assays, we detected the effect of c9, t11-CLA and COX-2 on the cell migration. RESULTS: Compared to NS-398 group, 200, 100 micromol/L c9, t11-CLA significantly suppressed SGC-7901 cells invading into the reconstituted basement membrane (F = 14.309, P = 0.000; F = 19.005, P = 0.000). 200 micromol/L c9, t11 CLA significantly inhibited SGC-7901 cells adhering to laminin, fibronectin and Matrigel (F = 3.063, P = 0.021; F = 6.692, P = 0.001; F = 11.999, P = 0.000). The chemotaxis of SGC-7901 cells and inhibitory frequency were significantly decreased in the 200 micromol/L c9, t11-CLA group (F = 1.380, P = 0.276). CONCLUSION: c9, t11-CLA inhibits invasion, adhesion and chemotaxis of SGC-7901 cells, and the COX-2 plays an important role in the process. [ Key words] PMID- 18399128 TI - [Effects of fluoride on neural cell adhesion molecules mRNA and protein expression levels in primary rat hippocampal neurons]. AB - OBJECTIVE: To investigate the effects of fluoride on the growth and viability, and mRNA and protein expression levels of neural cell adhesion molecules (NCAM) in primary rat hippocampal neurons. METHODS: The growth and development, the rate of cell survivor, and the mRNA and protein expression levels of NCAM were measured by MTT, RT-PCR, and Western blot respectively after the hippocampal neurons were incubated with 20, 40, and 80 microg/ml sodium fluoride for 24 hours in vitro. RESULTS: As compared with the control group, the number of cells, the length and number of neuritis, and rate of cell survivor were significantly decreased in 80 microg/ml fluoride-treated group (P < 0.05). The mRNA expression levels of NCAM in 40 and 80 microg/ml fluoride-treated groups were significantly lower than that in the control group and decreased with the increasing fluoride concentration. Compared with the control group, the mRNA expression level of NCAM in 20 microg/ml fluoride-treated group was decreased, but the difference was not statistically significant (P > 0.05). The NCAM-180 protein expression levels in 40 and 80 microg/ml fluoride-treated groups, the NCAM-140 protein expression levels in all fluoride-treated groups, and NCAM-120 protein expression level in 80 microg/ml fluoride-treated group were significantly lower than those in the control group (P < 0.05, P < 0.05, P < 0.05, respectively). CONCLUSION: Fluoride might restrain the growth and survival of rat hippocampal neurons, and decrease mRNA and protein expression levels of NCAM. The impairment of developmental hippocampus might be one of the neurotoxicant target sites for fluoride toxicity. PMID- 18399129 TI - [The establishment and assessment on cosmetic products using 3T3 mouse fibroblast neutral red uptake phototoxicity assay]. AB - OBJECTIVE: To establish the 3T3 mouse fibroblast neutral red uptake (NRU-PT) phototoxicity test method, and evaluate the practicality of the method in detecting potential phototoxicity of the cosmetic products. METHODS: Fifteen phototoxic and 9 non-phototoxic chemicals were tested in our laboratories, the phototoxic potential of the test chemicals was evaluated in a prediction model in which either the photo irritation factor (PIF) or the mean photo effect (MPE) was compared with the coherence and sensitivity of the method. 20 kinds of functional cosmetics were detected and the results were analyzed by the 3T3 NRU-PT in vitro and Guinea pig skin phototoxicity test (in vivo). RESULTS: Both PIF and MPE of the chemicals were highly reproduced, and the correlation between in vitro and in vivo data was almost perfect. All the non-phototoxic provided a negative result, while 14 of the 15 phototoxic tested chemicals gave clear positive results. For cosmetics, the correlation between in vitro and in vivo data was consistent. CONCLUSION: The 3T3 NRU PT test was established successfully, it should be used as a good alternative method for assessing the phototoxic potential of the chemicals and cosmetics in China. PMID- 18399130 TI - [Genotypes, allele frequencies and dynamic distribution on resistance-associated esterase genes of Culex pipiens complex in Hangzhou]. AB - OBJECTIVE: To investigate the genotypes , allele frequencies and dynamic distribution on resistance associated esterase genes of Culex pipiens complex in Hangzhou. METHODS: The PCR-restriction fragment length polymorphism (PCR-RFLP) assay was applied to type the resistance associated esterase genes, and dynamic surveillance on frequencies of the resistance associated esterase gene of natural population of Culex pipiens complex in Hangzhou during 2003-2005, and phenotype of the resistance associated esterase genes were detected by esterase starch gel electrophoresis technique. RESULTS: The PCR-RFLP assay of esterase allele genes for three consecutive years disclosed four esterase genotypes, namely, the world wide highly active homozygous Est beta 1(1) (50%-54%), homozygous Est beta 2 (29% 34%), heterozygous Est beta 1(1)/beta 2 (5%-10%) and Est beta N (3.13%) of a new homozygous genotype. The research of the resistance associated esterase genes phenotype in natural population of Culex pipiens complex in Hangzhou in 2005 with esterase starch gel electrophoresis technique revealed four major types, namely, Est beta 1(1) (61%), Est alpha 2/beta 2 (12%), Est alpha 8/beta 8 (7%) and sensitive phenotype (29%). CONCLUSION: There should be various resistance associated esterase genotypes in natural population of Culex pipiens complex in Hangzhou. During the period of 2003-2005, Est beta 1(1) was the major type; Est alpha 2/beta 2 was the second. Est beta N was a new esterase genotype detected in 2005 only with a mere percentage of 3.13%. As for its resistance to the new insecticide, a follow-up study should be needed. The molecular typing of the amplified esterase gene should be consistent with the resistance associated esterase genes phenotype. PMID- 18399131 TI - [The relationship of ankle brachial index to all-cause and cardiovascular disease mortality in Chinese male patients with hypertension]. AB - OBJECTIVE: To evaluate the risk factors for peripheral arterial disease (PAD) and the relationship of low ankle brachial index (ABI) to all-cause and cardiovascular disease (CVD) mortality in Chinese male patients with hypertension. METHODS: The data of 1606 male participants with hypertension from the eight hospitals in Beijing and Shanghai were analyzed. ABI was ascertained at baseline by measuring the systolic pressures on bilateral brachial and tibial arteries. ABI < or = 0.9 was used as the diagnostic criteria for PAD identification. The follow-up survey was conducted from November 2005 to January 2006. RESULTS: Of 1606 male participants with hypertension at baseline, 406 (25.3% ) were in low-ABI group and 1200 (74.7%) were in normal-ABI group. Older age, TC, history of diabetes, history of smoking and 2-grade hypertension were associated with low ABI in male patients with hypertension. During the (12.87 +/- 2.94) months follow-up, there were 153 deaths. Of which, 62 were attributable to CVD. Low ABI was associated with adjusted all-cause and CVD mortality risk of 1.728 (1.223-2.441) and 2.388 (1.409-4.046) respectively in Cox regression models. Rate of survival for the low-ABI group was significantly worse than for the normal-ABI group. The risk of all-cause and CVD mortality was increased with the decline of ABI. CONCLUSION: Low ABI is independently associated with the high risks of all-cause and CVD mortality in Chinese male patients with hypertension. The utility of ABI as a tool for predicting mortality in the patients with hypertension should be popularized. PMID- 18399132 TI - [The effects of sexually transmitted infections (STI)/AIDS behavioral intervention among female sex workers]. AB - OBJECTIVE: To explore the effective sexually transmitted infections (STI)/AIDS intervention strategies among female sex workers at the metropolis areas with low prevalence. METHODS: A comprehensive intervention was carried out among female sex workers in eight recreation establishments which have representative characters in Chaoyang district, and the effects of intervention was subsequently evaluated by a questionnaire from September 2005 to February 2006. The data were analyzed by SPSS11.5. RESULTS: The study subjects aged 17-40 year old (25.22 +/- 5.81), and 203 (89. 82%) of them had high or middle school education background. 166 were unmarried, and 205 (90.70%) had no job, and 244 (99. 12%) subjects were migrants. After intervention, the correct answer rates for the knowledge of STI/ AIDS rose from 52.6% to 75.4% (chi2 = 22.701, P < 0.01); and the correct use of condom were higher than those of baseline, with the rise from 22.12% to 34.76% (chi2 = 8.14, P < 0.05). CONCLUSION: The STI/AIDS comprehensive intervention measures were effective and should be evaluated and generalized in the similar metropolis regions. PMID- 18399133 TI - [Application of time series analysis in the prediction of incidence trend of influenza-like illness in Shanghai]. AB - OBJECTIVE: To forecast incidence trend of influenza-like illness in Shanghai. METHODS: We collected everyday-report influenza-like illness surveillance information from January, 2004 to April, 2006 and used autoregressive integrated moving average model (ARIMA) to analyze and establish prediction model. 114 weeks preceding information was used to establish model and 9 weeks data to evaluate. RESULTS: Model ARIMA (1,0,0) (1,1,0) 26 from Surveillance information was both with seasonal and non-seasonal features (P < 0.001). White noise analysis show the minimum Box-Ljung value of autocorrelation function was 0.803 (P > 0.1) and the residual was randomized difference. We established prediction model as lgY(t) = 0.879 lgY(t-1) + 0.418 lgY(t-26) - 0.367 lgY(t-27) + 0.582 lgY(t-52) - 0.512 lgY(t-53) and forecasting effect was well. True values were all between 95% CI of predicted ones. CONCLUSION: ARIMA model can be well used to simulate incidence trend of influenza-like illness in Shanghai. PMID- 18399134 TI - [Epidemiology of HPV infection and cervical cancer and perspective of HPV vaccination in Chinese women]. PMID- 18399135 TI - [Study on the distribution of human papillomavirus types in cervix among Chinese women: a meta-analysis]. AB - OBJECTIVE: To explore the distribution the tyes- of human papillomavirus (HPV) in cervical biopsy tissue or exfoliated cell specimen of Chinese women and to estimate the perspective of HPV prophylactic vaccine in China. METHODS: A total of 18 studies using polymerase chain reaction to detect HPV types were included in this Meta-analysis. There were 1335, 394, 381 and 2584 cases included in invasive cervical cancer (ICC), high-grade squamous intraepithelial lesions (HSIL), low-grade squamous intraepithelial lesions (LSIL) and normal group respectively. The overall HPV prevalence was estimated by adjusting the variables using unconditional multiple logistic regression model. HPV type-specific HPV prevalence rates of 18 types including HPV6, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 70, 73 and 82 were estimated, after stratified by cervical lesion of Chinese women. RESULTS: In ICC, HSIL, LSIL and normal group, the overall HPV prevalence rates were 82.7%, 88.5%, 69.3% and 13.1% respectively, and the HPV16 type-specific prevalence were 61.9%, 46.7%, 21.0% and 3.1%. HIPV16 was the predominant type in all cervical lesion specimen followed by HPV18 and 58 in ICC group, then HPV58 and 52 in HSIL, LSIL and normal. The estimated HPV16/18 positive fractions were 69.6%, 59.1%, 32.3% and 4.4% respectively in ICC, HSIL and normal. Data from the Meta-analysis showed that the top five common HPV types in ICC women were HPV16, 18, 58, 52 and 31 which contributed for 82.9% of the ICC. Thus the HPV16, 18, 31, 33, 35, 45, 52 and 58 were accounted for 87.7% ICC in China mainland. CONCLUSION: The proportional impact of HPV16/18 vaccine on ICC was estimated as high as 69.6% in China. HPV58 seemed the priority HPV type and should be emphasized enough to be included in the development of new generation HPV vaccine. However, more uniform standardized multi center studies were necessary to get more exact figure about HPV distribution in China. PMID- 18399136 TI - [Study on the correlation between human papillomavirus infection and cervical lesion among women living in community of Beijing]. AB - OBJECTIVE: To observe the correlation between human papillomavirus (HPV) infection and cervical lesion among women living in community of Beijing. METHODS: A total of 795 women at age 20-54, living in Zhanlanlu District of Beijing were screened for cervical lesion. Samples of cervical cytology (LCT) and HPV test (hc2) were collected. Colposcopy and biopsy were conducted in women with positive LCT. RESULTS: In those 795 women, the infection rate of HPV was 14.1% (112/795). In 40 women who were LCT positive 1 early invasive cervical cancer, 4 cervical intra-epithelial neoplasia (CIN3), 3 CIN2 and 7 CIN1 were noticed. In 750 women with negative LCT, 5 CIN1 and 1 low-grade CGIN were diagnosed. In those women who were Cyto(+) and HPV(+), 15 cases (55.6%, 15/27) were diagnosed with > or = CIN1 (including 7 CIN1, 3 CIN2, 4 CIN3 and 1 early invasive cancer). CONCLUSION: The risk of cervical lesion significantly increased in women showing positive in cytology and HPV test. PMID- 18399137 TI - [Human papillomavirus infection and cervical intraepithelial neoplasia morbidity of women from different occupations in Shenzhen city, China]. AB - OBJECTIVE: To investigate the human papillomavirus (HPV) infection and cervical intraepithelial neoplasia (CIN) morbidity of women from different occupations in Shenzhen city. METHODS: 2045 women of five kinds of occupation in Shenzhen city, including 130 teachers, 385 workers, 316 service women, 199 poverish women, 420 doctors or nurses and 595 general residents were included. We screened these women by methods of detecting high risk HPV of hc2 combing with LCT. Women with screening positive results were diagnosed CIN by colposcopic biopsy. RESULTS: (1) High risk factors on HPV infection rate in different occupations were different with the highest in service occupation (19.3%) while the lowest appeared in medical workers (11.9%). (2) In those 2045 women, we found 199 cervical lesions including pathological HPV infection, CIN1, 2, 3 and cervical cancers, with morbidity rates as 4.11%, 3.28%, 1.67%, 0.54% and 0.15% respectively. Along with the progress of the cervical lesions, the morbidity decreased. (3) The morbidity rates of CIN in different occupations were different, with the highest of HSIL in service occupation and the lowest in teachers. CONCLUSION: Women of different occupations in Shenzhen city had different high risk HPV infection rates and CIN morbidity. The HPV infection rate and HSIL morbidity were highest among women having service related jobs. PMID- 18399138 TI - [Study on the prevalence of human papillomavirus infection and distribution of types in Shenyang city]. AB - OBJECTIVE: To study the status of infection and type-specific distribution on human papillomavirus (HPV) in the urban areas of Shenyang city, China and to provide data for vaccine introduction. METHODS: A list of 1000 women aged 15-59 years were obtained randomly from the local population, and a questionnairey was filled in. Samples of cervical cell were obtained from 702 women who had had sexual experience and were not pregnant during the study was carried on. HPV type was detected using a GP5+/6+ -based PCR, liquid-based cytology, VIA, VILI and electronic coloscopy were used for screening on cervical cancer. RESULTS: The prevalence of any HPV type was 17.0% among the 702 women who had been enrolled in this study, 64.1% in women with abnormal cytology, 11.1% in women with normal cytology and the difference was significant. The prevalence of high-risk HPV type was 11.9% and the low-risk type was 6.5%. The prevalence of either high-risk or low-risk HPV type was significantly higher in women with abnormal cytology than those with normal cytology. There were a total number of 32 HPV types identified and the most common HPV types were 16 (3.4%), 52 (2.4%) and 58 (1.9%). The prevalence rates of either high-risk or low-risk HPV type were not statistically different among different age-groups with P>0.05. HPV types with high risk were predominantly distributed in the groups below 45 years old and the difference was significant with P < 0.05. There was no significant difference between high-risk and low-risk types above 45 years old. CONCLUSION: The prevalence of HPV in women with abnormal cytology was statistically higher than those with normal cytology. The most common HPV types in this population were 16, 52 and 58, and the validity of present vaccines was not good enough for cervical cancer prevention in this local area. PMID- 18399139 TI - [Study on cervical intraepithelial neoplasia in relation to human papillomavirus and co-factors: a case-control study in China and Australia]. AB - OBJECTIVE: This study was to elucidate the role of human papillomavirus (HPV) types and cofactors in the development of cervical intraepithelial neoplasia (CIN). METHODS: Two hundred and twelve women with CIN and 427 women with normal cervical cytology (control group) were recruited from China and Australia. A questionnaire was administered to each participant to obtain the demographic and risk factor information. Cervical biopsies or smears were taken to detect HPV DNA by PCR and to identify HPV types by direct sequencing and/or Amplicor hybridisation. Data were analyzed by logistic regression. RESULTS: HPV prevalence rates of specimens from Chinese and Australian were 11% and 15% among controls (P >0.05), with 99% and 85% of CINs (P<0.001), respectively. The presence of any type of HPV DNA was strongly associated with CIN with OR 43.3 for Chinese and OR 541.6 for Australian women. The strongest risk was for HPV16,followed by HPV31 in Australians, but HPV58, 59 in Chinese women. The risk for multiple HPV infection was stronger in the Australians than that in the Chinese cohort. Except for HPV infection, educational attainment was unexpectedly associated with an increased risk for CIN in Chinese, and cancer history in family was a risk factor for Australians. For the combined cohorts, educational attainment, and frequency of vitamin consumption were identified to be risk factors for CIN. CONCLUSION: Cervical HPV DNA was a major risk factor, with the highest relative risk for type 16 HPV infection for CIN. There were variations in the distribution of HPV genotypes and cofactors in China versus Australia and in CIN. PMID- 18399140 TI - [Study on the value assessment of various screening programs regarding cervical cancer screening strategy in the rural areas of China]. AB - OBJECTIVE: To evaluate the application value and feasibility of various cervical screening methods and to explore a rapid and efficient cervical cancer screening program for the women in the rural areas of China. METHODS: We sequentially conducted human papillomavirus (HPV) DNA test by hybrid capture-2 (hc2) with cervical cells, liquid-based thinprep cytology test (TCT), visual inspection with acetic acid (VIA), visual inspection with iodine (VILI), colposcopy respectively for the 2499 married women between 30 and 49 years from Xiushui county of Jiangxi province. All the detection methods were performed independently under double blind design. Women who were diagnosed positive for having any VIA,VILI and colposcopy inspection or for those women who were diagnosed negative for VIA, VILI and colposcopy but with positive result of HPV or TCT test underwent cervical biopsy directly and endocervical curettage (ECC)when necessary. We performed cervical biopsy endocervical curettage within two weeks to observe the sensitive (SE), specificity (SP), negative predict value (NPV) and positive predict value (PPV) of these detection methods when used alone or combined each other, including HPV test, TCT inspection, VIA, VILI, and colposcopy, the pathological diagnosises of cervical tissue were confirmed by IARC (International Agency for Research on Cancer) while the cytological findings were underegone through the updated program of TBS (The Bethesda System) in 2001. RESULTS: A total of 2499 women underwent the screening and found 443 women who were diagnosed as HPV positive, 337 women with abnormal cervical cytology and 27 women with ASC-H, 157 cases with ASCUS; 103 cases with HSIL, 49 cases with LSIL and 1 cervical cancer. According to the pathological findings. There were 181 women diagnosed as cervical intraepithelial neoplasia (CIN) or cervical cancer, including 81 cases with CIN1 37 cases with CIN2,60 case cervical cancer. The sensitivity rates of HPV, TCT, HPV+ TCT, VIA, VILI, VIA+VILI and colposcopy were 96.67%, 89.47%, 97.98%, 56.57%, 36.36%, 63.64% and 39.39%, and the specificity rates were 85.00%, 96.91%, 86.97%, 94.60%, 96.23%, 92.97% and 98.14% respectively. CONCLUSION: HPV + TCT seemed to be more sensitive than other screening methods in the cervical cancer screening program. PMID- 18399141 TI - [Analysis on the expression of human papillomavirus type 16 E2 and E6 oncogenes and disruption of E2 in cervical cancer]. AB - OBJECTIVE: To evaluate the roles of expression and early protein E2 and E6 load of human papillomavirus type 16 (HPV16) on cervical cancer in order to explore the relation between disruption of E2 and development of cervical cancer. METHODS: A case-control study was conducted, including 141 cervical cancer patients as cases who had been diagnosed by cytological approaches and histological approaches in Shanxi province Tumor Hospital, China. Two type of controls including 137 hospital controls with hysteromyoma by cytology or histology and eligible 129 controls from 1582 healthy women in the community who took part in community-organized physical examination with neither CIN2-3 nor invasive cancer, nor other gynecologic diseases were recruited. HPV16 E2 and E6 oncogenes were detected by multiple polymerase chain reaction (multi-PCR). The levels of E2 and E6 were analyzed used Bio-1D+ + software provided by VILBER pattern formatter. RESULTS: The positive rates of HPV16 E6 in cancer cases (46.8%) were significantly higher than that in hysteromyoma group (24.1%) or healthy control group (2.3%) and accounted for 2.77 of OR (95% CI: 1.66-4.63) and 36.96 of OR(95% CI: 11.22-121.71) respectively. The expressions and loads of HPV16 E6 and E2 in cases were significantly higher than that in two control groups. Meanwhile, the expression or level of E6 was higher than that of E2 in each group. Disruption rate of E2 was 22.73% and the ratio of E6 to E2 was 1.24 in cervical cancer group. CONCLUSION: The positive rates and levels of HPV16 E6 or E2 found in cervical cancer were higher than that in hysteromyoma and healthy women. High expression of E6 and disruption of E2 might play an important role in the development of HPV-induced cervical cancer. PMID- 18399142 TI - [Programs on epidemiological research of human papillomavirus]. PMID- 18399143 TI - [Study on the prevalence rate and risk factors of chronic obstructive pulmonary disease in rural community population in Hubei province]. AB - OBJECTIVE: To explore the prevalence of chronic obstructive pulmonary disease (COPD) and its risk factors in rural area in the southern part of Hubei province and to provide evidence for prevention and treatment. METHODS: Using uniform scheme, procedures and questionnaire, a multistage survey for 1883 people in rural area in southern part of Hubei province was performed, and physical examinations and lung function tests were conducted for every participant. RESULTS: In this survey, 186 COPD cases were identified with a overall prevalence of 9.88%. The prevalence for male was 13.68% while in female it was 6.46%. The prevalence was higher in males than in females (chi2 = 27.48, P< 0.001) and higher with the increase of age (chi2 = 79.22, P<0.001). Factors associated with COPD were identified in one-way variance model as follows: sex, age, smoking, time for cooking, using biomass fuels for in-door cooking, family history, frequent cough before age 14, low body index and having house remodelling in the last five years. Results from multivariate logistic regression analysis showed that sex (OR= 1.3010), age (OR = 1.8506), smoking (OR= 3.0118), cooking time (OR= 1.7651), family history (OR = 1.5278), frequent cough before age 14 (OR = 2.8965) were risk factors. CONCLUSION: The prevalence of COPD in the southern part of Hubei province was high, suggesting that comprehensive intervention measures should be taken. PMID- 18399145 TI - [Study on the prediction of smear negative pulmonary tuberculosis with classification trees]. AB - OBJECTIVE: To improve the respiratory isolation policy for patients with suspected pulmonary tuberculosis (TB). METHODS: All consecutive patients with suspicion of having pulmonary TB when seeking health care at the County TB dispensary of the Center of Disease Control and Prevention received face to face interview. RESULTS: A Classification model was constructed with a sensitivity of 90.9% and specificity of 90.2%, while predictive factors of culture-proven pulmonary TB among smear negativecases were soakage in Chest X-ray exam (77.0% vs. 4.4%; P<0.0001), bilateral lung's abnormal (1.6% vs. 19.4%; P<0.0001) and reaction of tuberculin skin testing (0.0% vs. 2.6%; P=0.014). CONCLUSION: Soakage, bilateral lung's abnormal and positive reaction of tuberculin skin testing were important predictors to prognosticate culture positive diagnosis. The model had been proved to have promising sensitivity and specificity in the rural population covered by NTP-DOTs. PMID- 18399144 TI - [A survey on the infection rate of tuberculosis among employees of the antituberculosis institutions in Henan province, China]. AB - OBJECTIVE: To study the situation of tuberculosis (TB) infection among the employees of the anti-TB institutions in Henan. METHODS: Cross-sectional study was adopted the employees working in all municipal-level- anti-TB institutions and 40 anti-TB institutions at county-level selected randomly from 109 counties of the province were regarded as surveyed objects. Tuberculin skin test (TST) was used to test the infection with PPD. RESULTS: 2153 employees accepting the TST and the positive rate was 60.6%, of which the positive rate was 66.1% among healthcare workers. Among the employees and healthcare workers, the positive rates of TST adjusted by the stratum weights between municipal-level and county level institutions were 57.3% and 62.8% respectively with Chi-square test the analysis of multivariate logistic vegression, both positive rate and strong positive rate among healthcare workers, the employees older than 30 years of age and working in municipal-level institutions were significantly higher than those among non-healthcare workers, the employees younger than 30 years old and working in county-level institutions, respectively. There were not significant differences of positive and strong positive rates between employees with and without BCG-history, or between male employees and female employees. CONCLUSION: Program on TB infection control in anti-TB institutions of Henan were weak and the employees especiolly healthcare workers had a high vocational exposure. PMID- 18399146 TI - [The impact of social norm and susceptibility on drug use among adolescents]. AB - OBJECTIVE: To investigate the role of knowledge, attitude and perceived social norm on the use of New Type of Drugs (NTD) and to construct the mediated model among these factors. METHODS: With stratified random group sampling, 3018 students from 3 middle schools, 3 high schools, 3 vocational high schools and a municipal-own university in Wuhan city completed questionnaires under anonymous and voluntary practice. A conceptual model was developed based on Reasoned Behavior Theory. Structural equation modeling (SEM) was used to test the proposed model. RESULTS: The overall prevalent rate of NTD was 3.28%, with 4.81% for males, 1.94% for females,1.80% for middle school students, 2.91% for general high school students, 7.83% for vocational school students and 3.25% for college students. SEM seemed to have been fixed nicely into the model. Adolescent NTD was significantly predicted by knowledge, attitude, perceived social norm which were all directly affecting or medicated by the susceptibility of drugs. CONCLUSION: Knowledge, attitude, perceived social norm might increase the risks for NTD use among adolescents, suggesting that preventive education on drug use among adolescents should be paid to improve the knowledge on drugs. PMID- 18399147 TI - [Study on molecular epidemiology of people infected with human immunodeficiency virus-1 in Hubei province]. AB - OBJECTIVE: To study the distribution of human immunodeficiency virus-1 (HIV-1) genotypes in Hubei province. METHODS: Epidemiological survey was carried out to HIV-1 carriers who were identified in Hubei province. HIV-1 env V3-V4, gag P17/24 and the first exon of tat region were amplified by nested-polymerase chain reaction(nPCR) .The sequences were determined, and phylogenetic analyses were then performed. RESULTS: 4 HIV-1 strains or circulating recombinant forms (CRFs) were identified in Hubei province with subtype B' the predominant which covered 5 kinds of populations including former blood donors, blood receivers, spouses of the infected people, sex workers and their clients, homosexuals, mainly distributed in the areas with many former blood donors. CRF08-BC and CRF01-AE were found distributed in economically more developed cities or southern area of the province, and the major transmission routes was through sexual contact. Only 1 patient, an injecting drug user, was identified having subtype C. CONCLUSION: Subtype B' was the main epidemic subtypes in Hubei province while CRF08-BC, CRF01 AE and subtype C were also circulating in the province, indicating the transmission of the disease might to become more complex. PMID- 18399148 TI - [Study on the characteristics of Tsutsugamushi disease in the epidemic areas of south islands in China]. AB - OBJECTIVE: To study the increasing incidence and the characteristics of Tsutsugamushi disease in the areas of Nan Peng Lie islands, Nan Ao island, Wan Shan archipelago, Nao Zhou island and Lei Zhou peninsula, located in the southern part of China and to develop strategies for preventive measures. METHODS: Both epidemiological investigation, isolation and gene identification of Orientia tsutsugamushi, as well as pilot preventive measures were carried out. RESULTS: These islands belonged to the epidemic area of south subtropical zone of Tsutsugamushi disease. The main host was Rattus norvegicu and the overall rates of infection on Orientia tsutsugamushi were 22.78%-33.75%. The main biological vector was Leptotrombidium (Leptotrombidium) deliens and the rates of infection on Orientia tsutsugamushi were 40.00%-75.00%. 25 strains of Orientia tsutsugamushi had been isolated from Rattus norvegicu and Leptotrombidium (Leptotrombidium) deliens. Results showed that the isolated strains of Orientia tsutsugamushi were 15 Karp, 8 Kato, 2 Yonchon. Results from serological studies showed that the positive rate of anti-Orientia tsutsugamushi antibodies was high, in both residents and soldiers stationed in these islands. On these islands, rats and biological vectors were killed. Results showed that these measures had positive impact in reducing the incidence. CONCLUSION: Islands from the southern part of the country belonged to the epidemic area of Tsutsugamushi disease. People visiting this areas should be under protection. PMID- 18399149 TI - [Evaluation on the role of hepatitis A vaccine in the prevention and control of hepatitis A in Tianjin city]. AB - OBJECTIVE: To evaluate the effects of prevention and control strategies on hepatitis A. METHODS: Surveillance data on hepatitis A from 1990 to 2006 in Tianjin was analyzed, and the coverage rate of hepatitis A vaccine among targeted population was estimated, to compare the anti-HAV IgG level of children younger than 15 years old in 1999 and in 2005. RESULTS: Results showed that a) the morbidity of hepatitis A decreased from 25.26/10(5) in 1990 to 0.82/10(5) in 2006; b) the ratio of hepatitis A in viral hepatitis decreased from 30.43% in 1990 to 1.05% in 2006; c) the estimated coverage rate was 72.7%; d) the positive rate of anti-HAV among children younger than 15 years old in 2005 was distinctly higher than that in 1999. CONCLUSION: Positive results showed that it was successful to use hepatitis A vaccine as the strategy to prevent and control hepatitis A in the past five years in Tianjin. PMID- 18399151 TI - [Identification of a new subtype of blaADC produced by Acinetobacter baumannii isolated in children]. AB - OBJECTIVE: To investigate the genotype of blaADC which was a kind of AmpC produced by Acinetobacter baumannii (AB), isolated through the detection of 28 similar strains among children. METHODS: 28 strains of AB were collected and isolated from the Pediatrics clinic during 2006, and were identified through bacteria and susceptibility test using Vitex-32 automicroscan GNI and GNS cards. The genotype of blaADC was confirmed by polymerase chain reaction (PCR) and them sequenced. RESULTS: 3 of the 28 strains of AB showed multi-drugs resistance, with a positive rate of 10.71%. blaADC was discovered in 17 of the 28 strains and the positive rate was 60.71%. All the 28 strains of AB were resistant to Cefoxitin. blaADC positive strains were all sensitive to Ampicil/Sulbactam, and only one of them was resistant to Piperacillin/Tazobactan. There were no blaADC genes discovered in the strains that were resistant to Ampicil/Sulbactam or Piperacillin/Tazobactan. There were changes of amino acids on the site 4, 242, 342 and 376 in the sequence of blaADC of No.2 strain, comparing to gi /7258342/ emb /CAB77444. 1/ in GenBank. CONCLUSION: Above 60% of the AB isolated in children were carrying blaADC while a strain was collected from them at random. When they were undertaken nucleotide sequence analysis, significant difference was found from the others that landed in GenBank, which identified itself as new subtype. PMID- 18399150 TI - [Study on the association of hand, foot and mouth disease and enterovirus 71/CA16 among children in Beijing, 2007]. AB - OBJECTIVE: To reveal the etiological agent of hand, foot and mouth disease in children in Beijing. METHODS: Throat swabs were collected from 6 infants and young children with hand, foot and mouth disease who visited the affiliated Children's Hospital from May to June 2007. Aspirated fluid from tracheal intubatton, serum and cerebral spinal fluid (CSF) were collected from a 9 years old girl (No.4243) having central neural system complication of severe hand, foot and mouth disease and admitted to the hospital from the Emergency Department. Throat swab and aspirated fluid were inoculated into the cell lines Hep-2, MDCK and Vero for virus isolation. RNAs were extracted by Trizol from 6 throat swab specimens and aspirated fluid, serum while CSF was from that severe case (No.4243). The gene fragment from 5' UTR of enterovirus was amplified from throat swabs and aspirated fluid by reverse transcription-polymerase chain reaction (RT PCR) with the primer pairs located at the untranslated region of all enterovirus. EV71 was identified by RT-PCR with the 2 and half primer pairs located at different parts of VP1 gene of EV71. The PCR products for VP1 encoding gene of EV71 from the specimens were sequenced and sequence analysis was performed by comparing those published VP1 genes of EV71. EV71 and CA16 specific primers were used to identify the isolates by RT-PCR and the sequences were directly determined from PCR products. RESULTS: Gene fragments with expected molecular weight were amplified from all 6 throat swabs and the aspirate by the primer pairs universal for the 5' UTR of enterovirus, suggesting that these patients with hand, foot and mouth disease were infected by entorovirus. Out of these 7 specimens, 2 throat swabs and the aspirate were also showing positive for the VP1 of the EV71 by different primer sets. Sequence analysis revealed that the sequences for the amplicons from 1 throat swab (No. F4211) and the aspirate shared highest homology with those published EV71, indicating that these specimens were truly positive for EV71. The sequences amplified from these specimens shared 100% and 98.9% homology to each other and were closer to the sequences of EV71 identified from Zhejiang province than those from Taiwan and strain BrCr. Gene fragments for 5' UTR of enterovirus were obtained by RT-PCR after CPE appeared in 6 out of 7 inoculations including that aspirate fluid in Vero cell, indicating that enteroviruses were isolated from these specimens. Virus isolates from one throat swab (No. F4211) and the aspirate (No. 4243) were positive by RT-PCR with the primer pairs for EV71, which was consistent with RT PCR amplification directly from specimens. Virus isolates from other 4 specimens were CA16 by RT-PCR and sequence analysis. CONCLUSION: These data suggested that hand, foot and mouth disease recently appeared in children in Beijing was related with EV71 and CA16. EV71 could cause severe clinical manifestations with central nerve system complications even in the child older than 5 years. PMID- 18399152 TI - [Isolation of hepatitis E virus from liver of swine at abattoirs in Shandong province]. AB - OBJECTIVE: To determine the prevalence and genotype of hepatitis E virus (HEV) in swine liver before on the market and to analyze the phylogenetic relationship between the isolates from swine and human. METHODS: 35 swine liver specimens were collected from two slaughtering houses in the countryside of Shandong province, China. Nested reverse transcription-polymerase chain reaction (nested RT-PCR) and subsequent sequencing were used to determine the nucleotide sequences. A phylogenetic tree was constructed with Neighbor-joining method based on the Kimura-2-parameter model. RESULTS: 3. (8.57%) of the 35 swine liver specimens being tested were positive for HEV RNA. The three swine HEV strains isolated in the present study from liver samples shared the highest identity to genotype-IV HEV. CONCLUSION: Results from the study confirmed that HEV was detectable among swine before on the market and the genotype was the same as that representing human and swine isolates in China. It also suggested that much more attention should be paid to the safety on the digestion of swine liver. PMID- 18399153 TI - [Study on ELISAs with rTpN17, rTpN47 and its fusion protein as antigens in the detection of serum samples from syphilis patients]. AB - OBJECTIVE: To construct the prokaryotic expression systems of tpnl7 and tpn47 genes and tpn17-tpn47 fusion of Treponema pallidum, and to establish ELISAs based on rTpN17, rTpN47 and rTpN17-TpN47 as antigens to evaluate the sensitivity and specificity of the ELISAs for detection of serological diagnosed syphilis. METHODS: tpn17 and tpn47 genes were amplified and cloned by routine molecular biological methods. PCR with linking primers was used to construct artificial fusion gene tpn17-tpn47. The prokaryotic expression systems of the genes were then constructed. SDS-PAGE was used to measure the expression of the target recombinant proteins rTpN17, rTpN47 and rTpN17-TpN47. Ni-NTA affinity chromatography was applied to extract the three recombinant proteins, while Western blot was performed to determine their immunity. Using rTpN17, rTpN47 and rTpN17-TpN47 as the coated antigens, ELISAs (rTpN17-ELISA, rTpN47-ELISA and rTpN17-TpN47- ELISA) were established to detect serum samples from 200 healthy individuals, 25 RA patients, 17 SLE patients and 419 syphilis patients. Results of the ELISAs were compared to those with TRUST and TPHA. RESULTS: The sequence similarities of the cloned tpnl7 and tpn47 genes and the constructed tpn17-tpn47 fusion gene were 100%, compared with the corresponding sequences in GenBank. The expression outputs of rTpN17, rTpN47 and rTpN17-TpN47 were 37.2%, 23.3% and 29.8% of the total bacterial proteins, respectively. Each of the three purified recombinant proteins showed a single fragment in gel after electrophoresis, and could take place remarkable conjugation reactions to the positive sera from syphilis patients. The detection results of rTpN17-ELISA, rTpN47-ELISA and rTpN17 TpN47-ELISA were negative for the serum samples from healthy individuals, RA and SLE patients, while presented 84.4%, 82.3% and 98.1% positive detection rates for the serum samples from syphilis patients. The positive detection rates of rTpN17 ELISA and rTpN47-ELISA were lower than that of TPHA (P<0.01), while the positive detection rate of rTpN17-TpN47-ELISA was similar to that of TPHA (P>0.05). All the positive detection rates from ELISA tests were higher than that of TRUST (71.4%). CONCLUSION: rTpN17-ELISA, rTpN47-ELISA and especially rTpN17-TpN47-ELISA established in this study were of great hope as it was rapid, simple, convenient, safe, with high sensitivity and specificity for serological screening and detection of syphilis. PMID- 18399155 TI - [Study on the association of clinical characteristic, CD4+ and level of HIV viral load among 690 initial HIV-infection]. AB - OBJECTIVE: To understand the correlation between CD4+ cell count, HIV viral load (VL) and clinical characteristics among patients when HIV-1 was tested positive and initial AIDS diagnosis was made. METHODS: 690 HIV-infected cases from Beijing Di-Tan Hospital were included and under a cross sectional study while SPSS statistical method was used. RESULTS: The 690 HIV-infected cases would include 458 males and 232 females with age range from 2-72 years (mean age as 35.3). The modes of transmission showed that: homosexual contact taking up 17.5% while heterosexual was 16.7%. Most of the homosexual-infected ones lived in Beijing and most of them had bachelor or master's degrees. 19.4% of the transmission happened between heterosexual/bisexual couples, suggesting that HIV was transmitted through the "bridge population" while the rest were infected by contaminated blood/plasma. Many of the cases were identified when they lately visited the pre operation surveillance point in the hospital. Serious immunodeficiency symptoms or signs were discovered as: CD4+ count < 50 cell/microl, serious opportunistic infections including pneumocystosis pulmonary, cerebral toxoplasmosis and cryptococcal meningitis. Higher frequencies of diseases seen were dermotosis, pneumonia, upper respiratory tract infection, hepatitis and digestive tract moniliasis. CONCLUSION: Because of the late identification of the disease, serious immuo-suppression situation often appeared, suggesting that there was an urgent need to improve STD/AIDS knowledge on those HIV (+) people so they might have an early access to accept medical care. PMID- 18399154 TI - [Evaluation on the accuracy of obese indexes in predicting obstructive sleep apnea-hypopnea syndrome in male adults]. AB - OBJECTIVE: To determine the best cutoff value and reference standard of obese indexes and to compare their diagnostic value in screening, predicting and diagnosing obstructive sleep apnea-hypopnea syndrome (OSAHS) in male adults by receiver operating characteristic curve (ROC). METHODS: A total of 1110 male adults were included and cross-sectional study was adopted. Four parameters including: body mass index (BMI), neck circumference (NC), waist circumference (WC) and waist-to-hip ratio (WHR) were studied. The correlativity between apnea hypopnea index (AHI) and BMI, NC, WC and WHR were analyzed by pearson correlation test. ROC curve was used to assess the diagnostic value and determine their best cutoff value to predict OSAHS and reference standard to screen/diagnose it. Their conformity in predicting OSAHS was analyzed by area under curve (AUC). SPSS software version 13.0 was used for statistical analysis and P< or =0.05 was considered to indicate statistical significance. RESULTS: (1) All of the fourindexes (BMI, NC, WC and WHR) showed a significantly positive correlation with AHI in adult male patients, with their Pearson coefficients as 0.373, 0.276, 0.291 and 0.127, respectively, and their P value were all below 0.001; (2) The best cutoff value of BMI, NC and WC in predicting OSAHS were 26.5 kg/m2 , 39 cm, 95 cm, and their corresponding sensitivity rates were 54%, 57% and 64% while rates of specificity as 73%, 65% and 53%; (3) BMI was better than other two indexes in predicting OASHS in adult male patients; (4) The value of BMI, NC and WC in screening OSAHS among adult male patients with 90% sensitivity and 10% rate of missed diagnosis as 23 kg/m2, 35 cm, 85 cm; (5) The value of BMI, NC and WC in screening OSAHS in adult male patients with 90% specificity and 10% misdiagnosis rate as 29 kg/m2, 43 cm, 105 cm, respectively. CONCLUSION: BMI seemed better in predicting OSAHS in male adults than NC, WC and WHR. The best cutoff values to predict OSAHS of BMI, NC and WC were 26.5 kg/m2, 39 cm and 95 cm, respectively. Their reference standards to screen OSAHS were 23 kg/m2, 35 cm and 85 cm while 29 kg/m2, 43 cm and 105 cm to predict OSAHS. PMID- 18399156 TI - [Study on the relationship between chronic glomerulonephritis and human leukocyte antigen in Shandong province]. AB - OBJECTIVE: The purpose of this study was to observe the association between chronic glomerulonephritis (CGN) and human leukocyte antigen (HLA) on DNA level in order to identify susceptible and protective genes and to further explain the possible pathogenesis of CGN. METHODS: 1073 renal transplantation patients with Han ethnicity were included in this study. All patients were recruited from three provincial Hospitals during the past ten years. The control group contained 7418 healthy Han volunteer donors from Shandong Hematopoietic Stem Cell Data Bank of China. We collected data about the polymorphism of HLA-I , II and DRB1. Gene frequency (GF), relative risk (RR) and correlation test were analyzed using statistical software. Some patients carrying the susceptible genes were followed up for 1,3 and 5 years, and compared their survival rate respectively. RESULTS: The frequency of HLA-A23, A25,B15, B40, B53 and DRB1 * 18 alleles increased significantly in CGN patients than in controls, showing that they might be the suspicious susceptibility genes of CGN. After the follow-up periods, the prognosis of patients with the susceptible genes was worse than the controls. The frequency of haplotypes of A23-B44-DRB1 * 18, A25-B15-DRB1 * 07, A3-B70-DRB1 * 11, A68-B13-DRB1 * 04, A11-B10-DRB1 * 12 increased significantly in CGN patients than in controls. There were 8 lower frequencies alleles (including A20, A22, A35, A36, A38, B21, B73 and B78) in CGN patients that were not found in the control group. The frequencies of the HLA-A32, A33, B50, B58, B60, B71, DRB1 * 16 alleles decreased significantly in CGN patients than in controls, showing that they might be the protective genes of CGN. CONCLUSION: Our data showed that there might be corresponding susceptibility and protective genes of CGN in Han population, in Shandong. There was significant association between the five common haplotype and CGN in Shandong population. However, the prognosis of the patients with the susceptibility genes was worse than the controls. PMID- 18399157 TI - [Identification of gene-gene interactions related to the etiology of complex disease: a multifactor dimensionality reduction-genotype pedigree disequilibrium test]. AB - To introduce the application of a multifactor dimensionality reduction-genotype pedigree disequilibrium test (MDR-PDT) for detecting gene-gene interactions in the etiology of complex disease. A brief overview on the basic theory, implementing steps and features of MDR-PDT were described, and a practical research case was demonstrated to application of MDR-PDT in nuclear family studies. The MDR-PDT approach was the extension or development of conventional MDR method which could be used for detecting gene-gene interactions in families of diverse structure. CONCLUSION: MDR-PDT was a new nonparametric and model-free method which might use additional family members in the nuclear families and had a good power to identify gene-gene interactions. PMID- 18399158 TI - [Pharmaceutical intervention for pandemic influenza]. PMID- 18399159 TI - [To investigate the carriage drug resistances and serotype of Haemophilus in the upper respiratory tract among healthy children]. PMID- 18399160 TI - [Study on polymerase chain reaction technology in the detection of Brucella DNA from blood samples]. PMID- 18399161 TI - [Study on occupational stress and breast cancer risk among professional women in China]. PMID- 18399162 TI - [A 1:2 matched case-control study of autoantibodies in esophageal cancer patients of Kazakhs ethnicity in Xinjiang]. PMID- 18399163 TI - [Analysis on current status regarding gynecological health examination of rural resident in Tianjin in 2004]. PMID- 18399164 TI - [Plague bacterium examination and epidemiology significance analysis of "Three Jiang Yuan Area" in Qinghai province, China]. PMID- 18399165 TI - [The analysis of data on case from out-patient clinic of dog-bite wounds in Ninghai country Zhejiang province 2005]. PMID- 18399166 TI - [About the treatment of multiple myeloma]. PMID- 18399167 TI - [Effect of CYP2C19 gene polymorphism on efficacy of thalidomide-based regimens for the treatment of multiple myeloma]. AB - OBJECTIVE: To study the distribution of different genotypes of CYP2C19 in multiple myeloma (MM), and investigate the effect of its polymorphism on efficacy of thalidomide-based regimens for the treatment of MM and discuss the role of antiangiogenesis in MM. METHODS: The CYP2C19 genotype of 92 patients with multiple myeloma was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The incidence of poor metabolizer (PM) in MM was compared with that in healthy Chinese people. After they were treated with thalidomide-based regimens, the response rate was compared between extensive metabolizers (EMs) and PMs. RESULTS: Of 92 patients, 18 (19.5%) were PMs, which was comparable to that in healthy ones. The response rates in EMs and PMs were 62.6% and 33.3%, respectively (P < 0.05). When patients were grouped by treatment regimens, the response rate in EMs was significantly higher than that in PMs (60.8% vs. 27.3%) for the thalidomide-dexamethasone group, and similar results were observed for the thalidomide-chemotherapy group (65.2% vs. 42.7%) though there was no statistical difference (P > 0.05). CONCLUSION: CYP2C19 genotype has no difference between MM patients and healthy person, but exhibits an effect on the treatment efficacy of thalidomide for MM. The lower response rate observed in PMs is possibly due to the reduced activity to inhibit angiogenesis by thalidomide. PMID- 18399168 TI - [Clinical features of multiple myeloma patients with extramedullary disease: a report of 40 cases from a single center]. AB - OBJECTIVE: To analyze the clinical and laboratory features and risk factors of multiple myeloma (MM) with extramedullary disease (EM) and its extraosseous localizations at diagnosis and during the course of MM. METHODS: The clinical features, survival rate and prognostic factors were retrospectively analyzed in 40 patients having EM from a total of 418 MM patients hospitalized in Changzheng Hospital from 1993 to 2006. RESULTS: Among the 40 patients, the first three localizations of EM involved soft tissue, pleura or peritoneum and central nervous system (CNS). Median duration of follow-up was 30 months. The median overall survival (OS) was 28 months. Twenty-five patients (6%) were found to have EM at diagnosis (group A), and their median OS was 16 months and 15 patients (3.6%) developed EM during the course of the disease (group B), and their expected median OS was 72 months. There was a significant difference between group A and B (P = 0.0045) for OS. Compared with those in group A, patients in group B had a higher percentage of plasmacytes (P = 0.022) and plasmablasts (P = 0.029) in bone marrow, and less advanced stage for international staging system (ISS) (P = 0.027). Log-rank univariate analysis showed that higher CRP level, higher serum LDH, Stage II and III for ISS, Hb < 110 g/L at diagnosis were poor prognostic factors. However, multivariate analysis with COX model showed none of them were statistically significant. CONCLUSION: EM tumors are not a rare manifestation of MM. Soft tissue in the commonest area involved. Higher serum CRP and LDH level, more advanced stage for ISS, anemia and having EM are poor prognostic factors of MM. PMID- 18399169 TI - [Monoclonal antibody against brain derived neurotrophic factor inhibits myeloma growth and angiogenesis in the xenograft NOD/SCID animal model]. AB - OBJECTIVE: To evaluate the in vivo antitumor effect of anti-brain derived neurotrophic factor (BDNF) monoclonal antibody (MoAb) on a human myeloma xenograft animal model. METHODS: The xenograft tumor model was established in the nonobese diabetic/severe combined immunodeficiency (NOD/ SCID) mice by subcutaneous injection of human myeloma cell line RPMI 8226. The antibodies were injected intraperitoneally at a dose of 20 microg/mouse at day 1, day 2, day 3 after tumor cell inoculation or at a dose of 100 microg/mouse once a week after tumors were developed. The histologic and cytologic examination were performed to confirm the development of plasmacytomas. The microvascular densities (MVD) in tumors were analyzed by immunohistochemistry. The effect of anti-BDNF MoAb on the proliferation of RPMI 8226 cells in vitro and on endothelial cell network formation in the co-culture system were determined by 3H-thymidine incorporation assay and Matrigel network formation assay, respectively. RESULTS: The xenograft NOD/SCID animal model had high capacity for growth of RPMI 8226 subcutaneous tumors and presented pathologic features of plasmacytomas. After subcutaneous injection of RPMI 8226 cells, all mice developed localized tumors in (20 +/- 2) d. On 20 microg anti-BDNF MoAb 3 consecutive treatment, the mean tumor-free time was extended to (30 +/- 6) d and survival was significantly prolonged compared with IgG-treated group [(57 +/- 7) d vs (48 +/- 4) d, P < 0.05]. When mice died naturally, the tumors size in anti-BDNF MoAb treated ones was also reduced compared with control group [(157.9 +/- 21.6) mm3 vs (405.5 +/- 35.2) mm3, P < 0.05]. When the antibody treatment (100 microg/mouse) underwent from 27 th to 60 day once a week after tumor inoculation, the local tumor growth was inhibited partially and necrosis and infiltration were observed in the tumors. The median MVD in the antibody-treated mice (100 microg/mouse) was 11 vessels/0.216 mm2. The IgG treated mice had no decrease in MVD of subcutaneous tumors compared with untreated mice. In vitro, anti-BDNF MoAb (1.5 microg/ml) significantly but partially inhibited HUVEC network formation induced by RPMI 8226 (68.2% reduction) and significantly inhibited RPMI 8226 proliferation, too. The IgG (1.5 microg/ml) treated mice had no significant effect on both of two assays. CONCLUSIONS: Anti-BDNF monoclonal antibody could inhibit growth and angiogenesis in subcutaneous myeloma tumors. BDNF is a potential therapeutic target in MM. PMID- 18399170 TI - [Analysis of overexpression of vascular endothelial growth factor-C in patients with angioimmunoblastic T-cell lymphoma]. AB - OBJECTIVE: To explore the vascular endothelial growth factor-C (VEGF-C) expression and its clinical significance in malignant lymphoma. METHODS: Lymphoma cells were isolated by laser microdissection. VEGF-C expression in lymphoma tissue and microdissected lymphoma cells was measured by realtime quantitative PCR. Meanwhile, vessel ultrastructure was identified by transmission electron microscopy. RESULTS: Comparing with that in 8 patients with reactive lymphocyte hyperplasia, VEGF-C was overexpressed in angioimmunoblastic T-cell lymphoma, both in lymphoma tissue (n = 18, P = 0.0020) and in microdissected lymphoma cells (n = 10, P < 0.0001). Increased VEGF-C level was associated with bone marrow infiltration (P = 0.0039), skin involvement (P = 0.0046) and high-risk international prognostic index (P = 0.0302). In VEGF-C overexpressed cases, ultrastructural study showed dystrophic vessels, with swelling endothelial cells and absence of pericytes. CONCLUSION: The value of VEGF-C expression might be a biomarker of disease progression in angioimmunoblastic T-cell lymphoma. PMID- 18399171 TI - [Analysis of prognostic factors in 74 cases of diffuse large B-cell lymphoma]. AB - OBJECTIVE: To explore the relationship of clinic features, lab findings, the origin of tumor cell as well as prognosis in Chinese patients with diffuse large B-cell lymphoma( DLBCL). METHODS: Seventy four cases of primarily diagnosed DLBCL were analyzed. Immunohistochemistry stain was used to check the expressions of Bcl-6,CD10 and MUM1. RESULTS: Among the 74 patients, the average age was 58.5 years, the ratio of male to female was 1.64:1. 23.2% (16/69) cases developed in lymph node, 15.9% (11/ 69) in the extra node area. Among 55 follow-up cases, 13 (23.6%) died, and 12 (92.3%) died in the first year after diagnosis. The prognosis analysis showed that diagnosed at age > 65 years (P = 0.036), and the international prognostic index (IPI) (P = 0.009) were independent prognostic factors; origin of tumor cell had a trend to be a prognostic factor, but no statistic difference (P = 0.086). beta2-MG and Bcl-6 expression had no relation with the prognosis. CONCLUSION: The middle and old-aged male patients are the most common in DLBCL and the first-year mortality rate is higher. The age at diagnosis and IPI can predict the clinical outcome. The origin of tumor cell might suggest the prognosis. PMID- 18399172 TI - [Application of flow cytometric immunophenotypic analysis in the diagnosis of malignant lymphoma]. AB - OBJECTIVE: To explore the value of flow cytometric immunophenotyping (FCI) in the diagnosis and differentiated diagnosis of lymphoma and explain the immunophenotypic features and differences of malignant lymphoma. METHODS: Seventy four fresh samples of suspicious lymphoma were collected from Nov. 2004 to Aug. 2006. Each sample was individually evaluated by FCI. The results were analyzed and compared with the histopathological diagnosis. RESULTS: Among the 74 cases, the FCI data consisted with the final morphological diagnosis in 61 cases (82.4%). For the diagnosis of B and T non-Hodgkin's lymphoma (NHL), thymoma, carcinoma and benign lesions of lymph node, the concordance between FCI data and morphological diagnosis were 93.5%, 100%, 100%, 100% and 81.3%, respectively. CONCLUSION: Multi-parameter FCI analysis can provide important information and help for diagnosis of lymphoma. It is an assistant but necessary approach for the diagnosis and differential diagnosis of lymphoma. PMID- 18399173 TI - [Study on signal transduction pathway in differentiation and apoptosis of leukemia cells induced by heat shock protein inhibitor]. AB - OBJECTIVE: To explore the signal transduction pathway in the differentiation and apoptosis of leukemia cells induced by heat shock protein 90 (HSP90) inhibitor 17 Allyl amide-17-demethoxygeldanamycin (17AAG). METHODS: Kasumi-1 cells were treated with increasing concentrations or exposure time of 17AAG. The total kit protein (CD117), phosphorylated kit protein and its downstream signaling molecules were measured by Western blot analysis. Mutated kit protein from control and 17AAG-treated Kasumi-1 cells was immunoprecipitated and immunoblotted for associated chaperones. RESULTS: Total kit protein and kit activity were decreased in 17AAG treated cells, but c-kit mRNA level was not. Total AKT protein and phospho-AKT, as well as phospho-STAT3 were rapidly down-regulated in Kasumi-1 cell after treatment with 17AAG. There was no change in total STAT3 protein. Immunoprecipitation showed that 1 microM 17AAG treatment for 1 hour caused kit associated HSP90 decrease and HSP70 increase. CONCLUSION: 17AAG-induced apoptosis of Kasumi-1 cells is associated with a decline in Asn822Lys mutated kit protein level and phosphorylated kit, and with a downregulation in its downstream activated signaling molecules involved in proliferation. AKT is a client protein of HSP90. The changes of kit associated HSP90 and HSP70 satisfy the circulation mode of molecular chaperone complex. PMID- 18399174 TI - [Influence of tumor necrosis factor-alpha and interferon-gamma on erythropoietin production and erythropoiesis in cancer patients with anemia]. AB - OBJECTIVE: To explore impaired erythropoiesis and relative inadequacy of erythropoietin production in the anemic cancer patients and the correlation of tumor necrosis factor-alpha (TNF-alpha) or interferon-gamma (IFN-gamma) with inadequate erythropoietin (EPO) response and impaired erythropoiesis in cancer patients with anemia. METHODS: Fifty adult anemic and 15 non-anemic tumor patients were studied. Serum EPO levels were measured by radioimmunoassay (RIA) and serum soluble transferrin receptor (sTfR). TNF-alpha and IFN-gamma levels by enzyme-linked immunosorbent assay (ELISA). Log transformed EPO and sTfR values were used in statistical analysis. The R correlation analyses were performed. RESULTS: The mean serum immunoreactive erythropoietin level in anemic cancer patients [(23.11 +/- 10.00) IU/L] was not significantly higher than in healthy people (P = 0.053), but significantly lower than in IDA patients with similar degree of anemia [(43.00 +/- 22.00) IU/L, P < 0.01]. Both O/P EPO [0.88 (0.54 1.10)] and O/P sTfR [0.89 (0.57-1.22)] were significantly lower in anemic cancer patients than in controls and in non-anemic cancer patients. There was no significant difference between the latter two groups. Furthermore, the expected inverse linear relation between serum EPO and hemoglobin levels was absent in the anemic cancer patients, and so did the relation between serum sTfR and hemoglobin levels. There was no correlation between O/P EPO and O/P sTfR. The serum levels of both TNF-alpha and IFN-gamma in anemic cancer patients [(25.75 +/- 26.71) ng/L, (50.49 +/- 42.12) ng/L, respectively] were significantly higher than those in healthy controls (both P < 0.01) or in nonanemic cancer patients (both 0.01 < P < 0.05), and so did between non-anemic cancer patients and controls. The serum levels of TNF-alpha were inversely correlated with hemoglobin levels (r = - 0.40, P = 0.004), O/P EPO (r = -0.32, P = 0.025) or O/P sTfR (r = -0.36, P = 0.01); while serum levels of IFN-gamma were inversely correlated with hemoglobin levels (r = -0.36, P = 0.01) or O/P sTfR (r = 0.39, P = 0.006), but not with O/P EPO. Conclusions Anemia of cancer is due to impaired erythropoiesis and relative inadequacy of EPO production. TNF-alpha might inhibit EPO production and erythropoiesis, while IFN-gamma maybe directly inhibit erythropoiesis and be independent of EPO response inadequacy. PMID- 18399175 TI - [The risk factors for thrombosis, myelofibrosis and leukemia transformation in patients with polycythemia vera]. AB - OBJECTIVE: To reassess the natural history of polycythemia vera (PV) in Chinese and evaluate the relationship between the incidence of thrombosis, post polycythaemic myelofibrosis with myeloid metaplasia( PPMM) , leukemia transformation and the therapeutic outcome and prognostic factors. METHODS: The clinical manifestations, laboratory parameters and treatment were retrospectively analyzed in 287 patients with PV. Univariate analysis of prognostic factors was performed using Log-rank model and multivariate analysis using COX model in term of the incidence of thrombosis, PPMM, hematologic or non hematologic cancers and mortality. RESULTS: Of the 287 patients, the median follow-up time was 46 (8-360) months. 208 thromboses were recorded in 115 patients. Twice or more thrombotic events occurred on 59 patients (51.34%). Most of these episodes occurred either at presentation or in the 2 years before diagnosis. Elder patients, prior thrombosis, poor response to therapy were associated with poor prognosis. With these three adverse prognostic factors, the patients could be separated into different risk groups. The incidence of thrombosis was higher in high risk group. 36 patients progressed to PPMM, the median time to PPMM was 80 (7-190) months. Higher WBC count, splenomegaly and treatment with alkylating agent and hydroxy carbamide (HU) were associated with poor prognosis. 2 cases progressed to AML. 1 to lymphoma and 1 to nonhematologic cancer. 13 patients died, the cause of death was fatal thrombosis in 9 and AML in 2. CONCLUSION: The incidence of thromboembolism is higher and the time to myelofibrosis was shorter in Chinese PV patients than in western PV patients. The main factors that influence the survival of PV patients are thromboembolism and leukemia transformation. PMID- 18399176 TI - [Study of paired immunoglobin-like receptor B expression on dendritic cells and its relationship with immune tolerance in mouse]. AB - OBJECTIVE: To investigate the expression of paired immunoglobin-like receptor B (PIR-B) on dendritic cells (DCs) and its relationship with tolerogenic DCs (T DCs) in mouse. METHODS: DC2.4 cells, an immature dendritic cell line derived from C57BL/6 mouse, were stimulated by lipopolysaccharide (LPS) for 48 h to induce the mature dendritic cells (mDC) and cultured respectively with the recombined mouse interleukin-10 (rmIL-10) or recombined human transforming growth factor beta1 (rhTGF-beta1) to develop the tolerogenic dendritic cells (T-DC). Special small interference RNA (siRNA) molecular of PIR-B was chemically synthesized and transfected into DC2.4 cells (si-DC) by lip2000. The expression of PIR-B on DC2.4 cells was measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. The allogeneic lymphocyte proliferative capacity of DCs was measured by mixed lymphocyte reaction (MLR) using 3H thymidine incorporation test. The concentration of IFN-gamma in supernatants of MLR was analyzed by ELISA. RESULTS: Semi-quantitative RT-PCR and Western blot showed that PIR-B mRNA and protein were expressed on DC2.4 cells. RmIL-10 and rhTGF-beta1 induced the higher PIR-B mRNA and protein level on T-DCs (Relative values were 0.51 +/- 0.08 and 0.58 +/- 0.23; 0.85 +/- 0.07 and 0.87 +/- 0.14; 0.79 +/- 0.10 and 0.85 +/- 0.34, respectively) (P < 0.05). LPS down-regulated the PIR-B expression on mDC (0.35 +/- 0.10 and 0.32 +/- 0.04) (P < 0.05). The PIR-B mRNA and protein expression were inhibited by siRNA transfection (decreased by 78.9% and 74.2% respectively after 48 h interference) (P < 0.05). DC2.4 cells stimulated the proliferation of BALB/c mouse allo-genetic spleen cell. The mDC enhanced alloreactivity in MLR and the IFN-gamma secretion in supernatants. The T DCs alleviated the allo-genetic spleen cell proliferation (P < 0.05) and IFN gamma secretion in MLR (P < 0.05). Silence of the PIR-B expression (si-DC) also promoted of alloreactivity and enhanced the IFN-gamma secretion in MLR (P < 0.05). CONCLUSION: High expression of immune inhibition receptor PIR-B is one of the general features and molecular mechanism of dendritic cells to acquire immune tolerance in mouse. PMID- 18399177 TI - [Dear readers]. PMID- 18399178 TI - [Catastrophes--are they worse today than former?]. AB - When we were asked to write an article on the history of catastrophes and our perception of it, we quickly got up to a point were we understood that such a theme could easily be handled in form of an entire book. Because this is virtually impossible and because we physicians are only "guests" in the fields of meteorology, seismology and political sciences, this paper reflects more our personal view and experiences of this very complex topic. But like Heraklit who said: "Ta Panta rhei."--"everything flows" and who argued that the world is a permanent becoming and vanishing, in which everything constantly changes, we argue that not only the world changes but also our perception of catastrophic events. Only the future will tell how bad the today and the past has been. PMID- 18399179 TI - [The crash of a US Dakota in 1946]. AB - The crash of a US Dakota in 1946 in the alps of Bernese Oberland initiated an immense reaction of rescue. Aims, problems and challanges are the same today. PMID- 18399180 TI - [Tsunami in South-East Asia--rapid response deployment in Banda Aceh]. AB - On December 26, 2004 the second largest earthquake ever seismographically registered occurred in South-East Asia. It had a magnitude of 9.3 on Richter's scale and its epicentre was located on sea ground 160 km West of Banda Aceh, the capital of the province of Aceh on the island of Sumatra. The earthquake resulted in a tsunami which almost completely destroyed the city of Banda Aceh. Its death toll on the island of Sumatra was 168,000. The Swiss Humanitarian Aid Unit was deployed within a few days after the catastrophe with an advance team, which had to evaluate the need for supplies and personal in Banda Aceh. In close collaboration with relief forces of the Australian armed forces the team was able to deliver efficient medical and technical support. The most prevalent medical problems were: (1) Tsunami associated aspiration pneumonia; (2) Infected wounds of lower extremities; (3) Open bone fractures of lower extremities; (4) Tetanus infection. PMID- 18399181 TI - [Doctor's behavior in disasters]. AB - When facing a disaster outside of hospitals, physicians have to work in unfamiliar surroundings. Self-protection is the first priority: be aware of possible dangers of traffic, explosions, electricity, structural collapse and toxic gas. Arriving first, one should safe the scene of accident (e.g. breakdown triagle, turn off electricity) and try to get a general idea of what has occured. After that, call the rescue services (in Switzerland dial 144). Only once this ist done, should one begin treating patients. If the rescue services are at scene, look for the commander, usually fitted out with a warning west "Einsatzleiter" ("chef d'intervention" in the french speaking part of Switzerland). Introduce yourself (name, profession, skills and material). In disasters, fields other than medicine are often more important. Nevertheless, doctors can help with logical thinking, strong and agile hands and attentive listening. One should only do the tasks assigned. In situations with mass casualities, morbidity and mortality can be reduced by special measures. The most important for physicians is the Triage. Triage is the sorting of patients based on their need for treatment and transport with the available resources. Triage ist usually run by the most experienced doctor. PMID- 18399182 TI - [Important principles of war and disaster surgery]. AB - The actual situation in surgical training doesn't allow for intensive studies of war and disaster surgery anymore. There is a real risk for Switzerland to loose the know-how on these subjects in the future. With continuous surge in urban violence and increasing risks for technological and natural disasters, such knowledge is necessary to successfully manage such situations through optimal utilization of the available resources. PMID- 18399183 TI - [How hospitals should prepare for exceptional situations]. AB - Whenever case load grossly exceeds the available resources of a hospital, properly planned extraordinary measures are needed to boost performance. The crucial elements to be included in hospital disaster plans are: a well prepared alarm call system, simple lines of command with clear-cut responsibilities, measures to admit, triage and treat patients, a concept for internal and external communication including the needed tools. Such a disaster plan, based on an educated guess upon possible disasters and tailored to a realistic estimate of the hospital's resources, regularly needs to be checked and taught to the staff This article proposes guidelines how to successfully set up a hospital disaster plan. PMID- 18399184 TI - [Growing importance of the Coordinated Medical Services]. AB - Diminishing resources and growing risks call more than ever for optimum coordination of all medical assets between the different partners, so that all patients can be given the best possible medical support in all situations. The significance of the Coordinated Medical Services (CMS) has been underlined by the new ordinance which entered into force on 1 June 2005. PMID- 18399185 TI - [Nuclear, radiological, biological, and chemical dangers and risks]. AB - In contrast to military conflicts natural events, industrial disasters as well as terror attacks can surprise our society without warning. In all these events nuclear, radiological, biological or chemical dangers can be involved as well. The acute or retarded health effects on the public, first responders and society are well known. The psychological strain and disconcertion have also to be considered as well. Conditions to respond to these events with success are the early recognition and introduction of appropriate immediate measures. The early recognition is as such important because these events don't only have to start with an explosion. The early recognition should maybe base on the delayed appearance of atypical symptoms or with the increase of rare illnesses. The response to some incidence will inevitable includes drastical safety precautions such as evacuations, limitations of passenger traffic or food consumption. Precautions will only be carried out if they are understood by the public. Objective information and sensiblisation of the public and all concerned by this matter is crucial; without minimization or paranoia. PMID- 18399186 TI - [Biography changes in times of war and political turmoils]. AB - The principal therapeutical process in the case of refugees from the Caucasian region consists in sharing the traumatic experiences in a safe place with a person in which they can fully trust. As an extremely painful work this process is not a continuous one but very often interrupted and needs several sessions. PMID- 18399187 TI - [Special needs of women and children after a disaster]. AB - Disasters can have a severe impact on whole communities, but the impact can be especially difficult for the more vulnerable in our societies: women and children. This paper identifies a number of ways that women and children can be impacted, and explores some practical approaches that can serve to effectively address the special needs of women and children after a disaster. Specific community-based psychosocial practices are identified that can enhance the healthy psychological processes in children and women. This can be accomplished through the fostering of protective factors that strengthen what is known as "resilience processes". This is a new approach how to care to those who experienced disaster-related trauma or stress. It offers psychosocial methods that can be useful, practical and effective in post-emergency circumstances alternatively or integrated into standard medico-psychotherapy approaches where large populations of women and children are affected, particularly where people from cultures unfamiliar with western therapeutic concepts or interventions are affected. Possible applications and considerations for encouraging resilience processes in women and children who have experienced disasters are explored, along with a commentary about implications for applications in Switzerland. PMID- 18399188 TI - [Mental problems of victims and caregivers after critical incidents]. AB - Critical incidents often challenge the structures of a community to their upper limits. Therefore not only victims and their relatives, but often the whole community or region have to deal with disaster consequences, which have to be psychologically processed and fostering mental reactions, often far from normal experience. Meanwhile different rescue and security specialists are exposed to high pressure to perform and mental stress. In modern disaster deployments also specialists in psychological early interventions and caregivers are integrated, which are as well not immune to stress. Despite the chaotic initial position in the aftermath of disaster, a great deal of scientific evidence concerning acute stress reactions and possible long-term mental health consequences has been gathered. But chaos also holds hidden, often poorly measurable and therefore scientifically not comparable aspects, which might nevertheless be useful for understanding of and coping with psychological trauma. Therefore this article addresses not just scientific, evidence based medical and psychological aspects of traumatization but focuses especially on less measurable experiences from defined disasters. PMID- 18399189 TI - Constructing magnetic molecular solids by employing three-atom ligands as bridges. AB - The combination of some three-atom bridges with paramagnetic 3d transition metal ions results in the systematic isolation of molecular magnetic materials, ranging from single-molecule and single-chain magnets to layered weak ferromagnets and three-dimensional porous magnets. The design strategy and role of secondary components, such as co-ligands, templates and other mixed short ligands are discussed. PMID- 18399190 TI - Manganese(II)-lithium and -sodium inverse crown ether (ICE) complexes. AB - Extending to transition metals, the class of compounds known as inverse crown ethers, two mixed alkali metal-manganese(II) amide ring compounds with oxo cores have been synthesised and crystallographically characterised, together with an oxo-free alkyl-amido precursor. PMID- 18399191 TI - Biphasic hydroformylation in ionic liquids: interaction between phosphane ligands and imidazolium triflate, toward an asymmetric process. AB - Biphasic hydroformylation of dec-1-ene and styrene, at commercially competitive rates, can be successfully performed in imidazolium triflate ionic liquids; the ionic liquid network forms 'inclusion complexes' with the phosphane ligands used to modify the rhodium catalyst. PMID- 18399192 TI - Copper bromide complexed by fluorinated macroligands: towards microspheres by ATRP of vinyl monomers in scCO2. AB - We report the successful synthesis of poly(methyl methacrylate) (PMMA) by atom transfer radical polymerization using a catalyst ligated to a polymeric ligand having a dual role, i.e., the complexation of the copper salt and the stabilization of the growing PMMA particles; at the end of the polymerization, the catalyst is removed by supercritical fluid extraction leading to PMMA microspheres with low residual catalyst content. PMID- 18399194 TI - The mechanism of carbonate formation on Pd-Al2O3 catalysts. AB - Vibrational spectroscopic investigations of the adsorption of isotopically labelled and unlabelled CO and CO2 reveal that carbonate formation on Pd-alumina catalysts occurs via an "oxygen down" reaction of CO with hydroxyl groups on the support, whereas CO dissociation on Pd can be excluded. PMID- 18399193 TI - A dinuclear iron(II) complex, [(TPyA)FeII(THBQ(2-))FeII(TPyA)](BF4)2 [TPyA = tris(2-pyridylmethyl)amine; THBQ(2-) = 2,3,5,6-tetrahydroxy-1,4-benzoquinonate] exhibiting both spin crossover with hysteresis and ferromagnetic exchange. AB - Dinuclear [(TPyA)FeII(THBQ(2-))FeII(TPyA)](BF4)2 (1) possesses hydrogen bonding interactions that form a 1-D chain, and pi-pi interactions between the 1-D chains that give rise to a 2-D supramolecular-layered structure, inducing hysteresis in the spin crossover behavior; 1 has shown spin crossover behavior around 250 K with thermal hysteresis and ferromagnetic interactions at low temperature. PMID- 18399195 TI - The ionic introduction of an N1 unit to C60 and a unique rearrangement of aziridinofullerene. AB - A new chloramine-based aziridination of C60 and unique rearrangement of aziridinofullerene to azafulleroid is described. The ionic introduction of an N1 unit to C60 via an addition-cyclization mechanism was first achieved under mild conditions; the combination of chloramine and MS4A resulted in the promising rearrangement of aziridinofullerene to azafulleroid, and the isomerization could be performed catalytically. PMID- 18399196 TI - Photoresponsive rolling and bending of thin crystals of chiral diarylethenes. AB - Dithienylhexafluorocyclopentene with (R)- or (S)-N-phenylethylamide substituents formed rod-like and 0.2-1.0 microm-thick platelike crystals by sublimation; upon UV irradiation, the crystals bent concavely to the incident light and finally rolled crystals were obtained; the bent crystals were reconverted to flat crystals by visible light irradiation. PMID- 18399197 TI - Aligned carbon nanotubes with built-in FeN4 active sites for electrocatalytic reduction of oxygen. AB - The electrocatalytic site FeN4, which is active towards the oxygen reduction reaction, is incorporated into the graphene layer of aligned carbon nanotubes prepared through a chemical vapour deposition process, as is confirmed by X-ray absorption spectroscopy and other characterization techniques. PMID- 18399198 TI - A direct imaging of amphiphilic catalysts assembled at the interface of emulsion droplets using fluorescence microscopy. AB - An amphiphilic fluorescent catalyst Q9[EuW10O36] (Q = [(C18H37)2N+ (CH3)2]), assembled in the interface of emulsion systems, was directly imaged by fluorescence microscopy; the catalyst shows high selectivity and activity in the oxidation of alcohols using H2O2 as oxidant and the catalyst can be easily separated and recycled by demulsifying. PMID- 18399199 TI - Absolute asymmetric photocyclization in chiral diarylethene co-crystals with octafluoronaphthalene. AB - A photochromic diarylethene containing naphthyl groups formed a chiral crystal when co-crystallized with octafluoronaphthalene, although both molecules are achiral, and underwent highly enantioselective photocyclization owing to the conformational confinement in the crystal. PMID- 18399200 TI - Electrokinetic probes for single-step screening of polyol stereoisomers: the virtues of ternary boronate ester complex formation. AB - Electrokinetic probes based on the differential migration of ternary boronate ester complexes permit the selective analysis of micromolar levels of UV transparent polyol stereoisomers in urine samples via dynamic complexation capillary electrophoresis that is applicable to single-step screening of in-born errors of sugar metabolism, such as galactosemia. PMID- 18399201 TI - Structural features of the Cu(II) complex with the rat Abeta(1-28) fragment. AB - The interaction between Cu(II) and the rat amyloid beta (1-28) fragment in micellar solutions at pH 7.5 was investigated by CD and NMR spectroscopy; the proton-copper distances were used in restrained molecular dynamics simulations to obtain a structural model of the Cu(II) complex. PMID- 18399202 TI - Conformer independent heterodimerisation of linear arrays using three hydrogen bonds. AB - 5-Membered heterocycles are employed to give a conformer independent DDA array of hydrogen bonds, resulting in enhanced binding affinity to a complementary AAD array in comparison to a DDA array employing a 6-membered ring. PMID- 18399203 TI - Facile synthesis of a mesoporous silica-supported catalyst for Ru-catalyzed transfer hydrogenation of ketones. AB - A convenient method for preparation of a mesoporous silica-supported chiral catalyst by postgrafting a homogeneous catalyst on SBA-15 was developed and its application in the asymmetric transfer hydrogenation of aromatic ketones was investigated. PMID- 18399204 TI - Stepwise formation of "organometallic boxes" with half-sandwich Ir, Rh and Ru fragments. AB - Octanuclear complexes with half-sandwich Ir, Rh and Ru fragments and tetra(4 pyridyl)porphyrin (L1) and oxalate (L2) spacer ligands [(Cp*M)4L1]2[L2]4 (M = Ir (6a) M = Rh (6b)), [((cymene)Ru)4(L1)]2[L2]4 were prepared and characterized. PMID- 18399205 TI - Highly efficient submonolayer Pt-decorated Au nano-catalysts for formic acid oxidation. AB - A novel structure of catalyst, submonolayer Pt-decorated Au, has been synthesized with minimal use of Pt and shows markedly improved activity toward formic acid oxidation where it facilitates the direct oxidation of formic acid by suppressing the formation of poisonous species COads via the "ensemble" effect. PMID- 18399206 TI - Assembly of CdI2-type coordination networks from triangular ligand and octahedral metal center: topological analysis and potential framework porosity. AB - Reaction of a rigid triangular ligand 2,4,6-tris[4-(1H-imidazole-1-yl)phenyl] 1,3,5-triazine (TIPT) with Cd2+ ions affordedrare non-interpenetrating CdI2-type networks which display high thermostability and potential porosity; the topological character of the CdI2-type networks have been analyzed in comparison with three common uniform (4,4), (6,3) and (3,6) plane nets. PMID- 18399207 TI - A biporous coordination framework with high H2 storage density. AB - A biporous three dimensional (3,6)-connected coordination framework is reported which exhibits excellent H2 adsorption at low pressures (< or = 1 bar) with high H2 adsorption density. PMID- 18399208 TI - A coincident spontaneous resolution system for racemic 1,1'-binaphthyl-2,2' dicarboxylic acid and 1,2-diphenylethylenediamine induced by water. AB - Spontaneously resolvable conglomerate crystals are obtained by combining racemic (rac)-1,1'-binaphthyl-2,2'-dicarboxylic acid and rac-1,2-diphenylethylenediamine, and this spontaneous resolution system is induced by water which is present in the crystallization solvent. PMID- 18399209 TI - Luminescent europium nanoparticles with a wide excitation range from UV to visible light for biolabeling and time-gated luminescence bioimaging. AB - Silica-encapsulated highly luminescent europium nanoparticles with a wide excitation range from UV to visible light (200-450 nm) have been prepared and used for streptavidin labeling and time-gated luminescence imaging of an environmental pathogen, Giardia lamblia. PMID- 18399210 TI - Solvatomagnetic effect and spin-glass behavior in a 1D coordination polymer constructed from EE-azido bridged MnIII3O units. AB - The non-uniform chain manganese(III) complex constructed from EE-azido bridged MnIII3O units, [MnIII3O(Brppz)3-(MeOH)3(N3)] 2MeOH [1, Brppz = 3-(5-bromo-2 phenolate)-pyrazolate], is the first example of 1D azido-bridged coordination polymer showing both solvatomagnetic effect and spin-glass behavior. PMID- 18399211 TI - A two-step field-induced magnetic transition in spin-canted systems observed only for the CoII coordination polymer. AB - Two isostructural 1D compounds {[M3(hpdc)2(H2O)6] 2H2O}n (M = Mn, Co; H3hpde = 2 hydroxypyrimidine-4,6-dicarboxylic acid) were synthesized by the in situ hydrothermal reactions of 2-chloropyrimidine-4,6-dicarboxylic acid with MCl2 (M = Mn, Co) and NaOH; the MnII compound shows spin-canted antiferromagnetism, whereas the CoII compound exhibits the coexistence of spin-canting and a two-step field induced magnetic phase transition. PMID- 18399212 TI - Supramolecular enantiodifferentiating photoisomerization of cyclooctene with modified beta-cyclodextrins: critical control by a host structure. AB - Enantiodifferentiating photoisomerization of (Z)-cyclooctene included and sensitized by m-methoxybenzoyl-beta-cyclodextrin gave chiral (E)-isomers in up to 46% enantiomeric excess, which is the highest value ever reported for supramolecular photochirogenesis with analogous hosts, thus demonstrating the crucial role of the sensitizer-spacer moiety in supramolecular photochirogenic systems. PMID- 18399214 TI - Site specific self-cleavage of certain assemblies of G-quadruplex. AB - It is demonstrated that certain structural assemblies of G-quadruplex are capable of performing selfcleaving actions in a site specific fashion. PMID- 18399213 TI - Immobilization of heptyl viologens in DNA strands both to inhibit dimerization and to accelerate quasi-reversible electron transfer reaction. AB - Viologens, mono-molecularly immobilized in DNA grooves, show reversible colour changes without attenuation during potential switching after covering with ionic liquid type polymers. PMID- 18399215 TI - Nanostructured microspheres of MnO2 formed by room temperature solution processing. AB - Nanostructured microspheres of manganese oxide have been prepared by the solution processing ofmanganese(II) salts using 1,4,7,10-tetraazacyclododecane and utilizing a crystalline intermediate stage. PMID- 18399216 TI - A general gold-catalyzed direct oxidative coupling of non-activated arenes. AB - A gold-catalyzed mild and general oxidative homo-coupling of arenes using PhI(OAc)2 as the oxidant is described (13 examples, 31-81% yield). PMID- 18399217 TI - Novel fluorophore based on a multi-substituted olefin skeleton with enhanced three-photon absorption in the femtosecond regime. AB - A new multipolar fluorophore based on a multi-substituted olefin skeleton that possesses strong three-photon absorption and optical-limiting properties in the femtosecond regime has been designed and synthesized; this archetype suggests a new strategy to further optimize molecular structures toward enhanced nonlinear absorptivities based on known materials. PMID- 18399219 TI - Dalton transactions: providing the best services. PMID- 18399218 TI - Self-assembled monolayer electrode of a diiron complex with a phenoxo-based dinucleating ligand: observation of molecular oxygen adsorptionldesorption in aqueous media. AB - The phenoxo-based dinucleating ligand, 2,6-bis[bis(6-pivalamido-2 pyridylmethyl)amino-methyl-4-aminophenol (1), and its Fe2(II) complex, [Fe2(II)(1)(PhCOO)2](CF3SO3) (2), were prepared and 2 deposited on the Au surface (2/Au) is much more stable than in solution and exhibits redox behavior in aqueous media as well as reversible adsorption/desorption of oxygen at room temperature. PMID- 18399220 TI - Self-assembly in inorganic and hybrid systems: beyond the molecular scale. AB - The availability of well defined inorganic nanocrystals allows the construction of aligned structures with characteristic architectural elements in the nanometer range, the mesoscale. Contrary to alignment driven by external conditions and fields, we focus on strict "self-effects", where the organization is already encoded in the shape and mutual interaction potentials of the particles. This Perspective discusses the potential of this approach for generating valuable functional inorganic mesostructures. PMID- 18399221 TI - The surface functionalisation of gold nanoparticles with metal complexes. AB - The use of nanoparticles is now expanding well beyond the field of pure materials science. In particular, the modification of the surface functionality is playing a key role in catalysis, sensor applications and immobilisation of biomolecules. The use of coordinated metals on the outer surface of gold nanoparticles is the focus of this short article. PMID- 18399222 TI - A method for the synthesis of nickel(0) bis(carbene) complexes. AB - A new method leading to Ni(NHC)2 (NHC = IMes, IPri, SIPr(i), SIBu(t)) complexes in moderate to good yields, involves the reaction of NHC (pre-formed or generated in situ) with Ni(CH3)2(tmed), tmed = N,N'-tetramethylethylenediamine; in one case, the intermediate Ni[I(Me2)Pr(i)]2(CH3)2, I(Me2)Pr(i) = N,N'-diisopropyl-4,5 dimethylimidazol-2-ylidene, has been isolated and structurally characterised. PMID- 18399223 TI - Zwitterionic bis(phenolate)amine lanthanide complexes for the ring-opening polymerisation of cyclic esters. AB - The reaction of Sm{N(SiMe3)2}3 with the bis(phenol)amines H2O2N(R) (H2O2N(R) = RCH2CH2N(2-HO-3,5-C6H2(t)Bu2)2; R = OMe, NMe2 or Me) gave exclusively zwitterions Sm(O2N(R))(HO2N(R)). For R = OMe or NMe2 these were efficient catalysts for the ring-opening polymerisation of epsilon-caprolactone and D,L-lactide with a tendency to form cyclic esters; in contrast, no polymerisation was observed for R = Me. PMID- 18399224 TI - A versatile route to bifunctionalized silsesquioxane (POSS): synthesis and characterisation of Ti-containing aminopropylisobutyl-POSS. AB - A facile synthetic pathway leading to significant yields of bifunctional polyhedral oligomeric silsesquioxanes (POSS) with two different functionalities on the same cage structure, namely Ti-NH2POSS, is proposed in this manuscript. PMID- 18399225 TI - X-ray structure of [ReCl4(mu-ox)Cu(pyim)2]: a new heterobimetallic Re(IV)Cu(II) ferrimagnetic chain. AB - A new heterobimetallic Re(IV)Cu(II) compound has been prepared and its crystal structure determined by single-crystal X-ray diffraction; magnetic susceptibility measurements show that this compound behaves as a ferrimagnetic chain with significant antiferromagnetic interactions between Re(IV) and Cu(II) metal ions. PMID- 18399226 TI - Group 13 organoderivatives supported on a metallic oxide model. AB - The [{TiCp*(micro-O)}3(mu3-CH)] (1) metalloligand, (Cp* = eta5-C5Me5), coordinates in a 1:1 ratio to [AlMe3] or 9-BBN to give [{Me3Al}{(mu3-O)(mu O)2(TiCp)2(TiCp)3(mu3-CH)}](2) or [{(C8H14)B}(mu-H) {(mu3-O)(mu-O)2(TiCp*)3(mu3 CH)}](4), respectively, partial hydrolysis of 2 leads to the new hydroxo aluminium derivative [{MeAl} {(mu-OH)(mu3-O)}2{(mu-O)2(TiCp*)3-(mu3-CH)}2](3). PMID- 18399227 TI - 1-trimethylsilylphosphirane as a ligand and as a stable masked reagent for phosphirane. AB - 1-Trimethylsilylphosphirane, C2H4PSiMe3, has been prepared on a multi gram scale from P(SiMe3)3 via CICH2CH2P(SiMe3)2. C2H4PSiMe3 is readily susceptible to protonolysis forming the thermally unstable parent phosphirane, C2H4PH, in good yields. Reaction of C2H4PSiMe3 with fac-M(CO)3(CH3CN)3 (M = Cr, Mo) or [Fe(eta5 C5H5)(eta6-C6H6)](PF6) give rise tofac-M(CO)3(C2H4PSiMe3)3 and [Fe(eta5 C5H5)(C2H4PSiMe3)3](PF6) respectively. Protonolysis of the free or coordinated 1 trimethylsilylphosphirane readily causes P-Si cleavage to give rise to the parent C2H4PH or the respective complexes,fac-M(CO)3(C2H4PH)3 andfac-[Fe(eta5 C5H5)(C2H4PH)3](PF6) in situ. All new complexes are characterised by analytical and spectroscopic methods and the X-ray crystal structures of fac Cr(CO)3(C2H4PSiMe3)3 and fac-Mo(CO)3(C2H4PH)3 have also been determined. PMID- 18399228 TI - Synthesis, structure and magnetism of a new dimeric silicotungstate: K9Na2CU0.5[gamma-Cu2(H2O)SiW8O31]2.38H2O. AB - A new structurally distinct dimeric silicotungstate K9Na,Cu0.5[gamma Cu2(H2O)SiWO31]2.38H2O (1) has been synthesized and characterized by infrared spectroscopy, elemental analysis, and variable-temperature magnetic measurements. Blue needle-like crystals of 1 were obtained by reaction of K8[gamma-SiW10O36] with 2 equivalents of Cu(n) in a 0.5 M sodium acetate solution (pH 4.2) and subsequent addition of an equal volume of ethylene glycol. The structure of 1 was determined by single-crystal X-ray diffraction: final R1 = 3.41% based on 9709 independent reflections. The structure consists of two [gamma-Cu2(H2O)SiWsO3,1]6- Keggin-like units with the [Cu2O(H2O)(micro3-O)(micro2-O)] moiety of one unit bonded to the top of a [W2O6] moiety of the other unit. Magnetic susceptibility measurements indicate competing ferro- and antiferromagnetic intramolecular coupling between the four S = 1/2 Cu(II) centers in the cluster anion. PMID- 18399229 TI - A new family of octanuclear Cu4Ln4 (Ln = Gd, Tb and Dy) spin clusters. AB - The reaction of Cu(OAc)2 and Ln(OAc)3 (Ln = Gd, Tb and Dy) with 2-amino-2-methyl 1,3-propanediol (ampdH2) under solvothermal conditions has afforded a new family of isostructural octanuclear Cu4Ln4 complexes with the formula [Cu4Ln4(OAc)12(ampdH)8(OH2)2] (Ln = Gd (1), Tb (2) and Dy(3)) in good yield. Variable temperature magnetic susceptibility measurements reveal weak intramolecular exchange interactions for 1 and 2. Ferromagnetic coupling is observed for 1 and attributed to Cu...Gd interactions. In contrast, the magnetic susceptibility behaviour of 2 arises from a combination of intramolecular exchange interactions and the crystal field splitting of the (7)F6 ground state of the Tb(III) ions. PMID- 18399230 TI - Investigating the topochemical polymerization of aniline derivatives in Pb(II) borate scaffolds. AB - Three different aniline derivatives, namely m- and p-aminobenzoic acids and m amino benzenesulfonic acid, were sequestered in between layers of a borate-based coordination polymer, based on lead(II) tetrakis(imidazolyl)borate. The aniline derivatives pre-organized in the interlayer spacing of the coordination polymer, and the reactivity of these pre-organized anionic monomers in the crystalline state was studied. We found that thermally activated reactivity under ambient atmospheric oxygen promotes what appears to be polymerization, and that the most reactive species is m-aminobenzenesulfonic acid monomer due to its increased mobility within the layers of the polymer. PMID- 18399231 TI - Square versus tetrahedral iron clusters with polyoxometalate ligands. AB - Two new insoluble transition metal substituted phosphotungstates, (C2N2H10)11[{(B alpha-PW9O34)Fe3(OH)3}4(PO4)4Fe].38H2O(1) and K4(C2N2H10)12[(alpha PW10Fe2O39)4].30H2O(2), have been isolated by the hydrothermal reaction of [A alpha-PW9O34]9-, Fe(III) ions and ethylenediamine. Compound 1 has a tetrahedral symmetry and contains a Fe13 core built from the assembly of four Fe(III) trisubstituted [B-alpha-PW9O34]9- anions around a central disordered iron ion via four phosphato ligands. The anion in 2 can be described as a square of disubstituted [PW10O37]9- anions linked by Fe(III)-O-Fe(III) bridges. Magnetic measurements performed on 1 and 2 have shown the occurrence of antiferromagnetic interactions between the iron ions and have allowed the coupling constants between the magnetic centers to be determined. PMID- 18399232 TI - Theoretical and electrochemical studies on organometallic symmetrical schiff base complexes of Zn(II), Cu(II), Ni(II) and Co(II). AB - The electronic communication between two redox centres through a Schiff base complex has been investigated in a series of ethylenediimine-bis(1-ferrocenyl-1,3 butanedionate) complexes of Zn(II) 1, Cu(II) 2, Ni(II) 3 and Co(II) 4. Cyclic voltammetry experiments of 1 and 2 exhibit a unique two-electron reversible oxidation wave, whereas in the case of 3 and 4 two and three one-electron oxidation processes are, respectively, observed. These results suggest some electronic interaction between the iron atoms of the ferrocenyl groups. DFT calculations carried out on model complexes show that for all the studied compounds the removal of the first two electrons corresponds to the oxidation processes of the iron centres in the weakly coupled ferrocenyl termini. The electronic communication between the two iron centres increases on going from 1 to 4. Finally, a re-indexation of the bands observed in the UV-Visible spectra has been carried out using TDDFT calculations. PMID- 18399233 TI - Water-soluble and stable dinitrogen phosphine complexes trans-[ReCl(N2)(PTA H)n(PTA)4-n]n+ (n = 0-4), the first with 1,3,5-triaza-7-phosphaadamantane. AB - The first dinitrogen complexes with the hydrosoluble PTA ligand, or its protonated form PTA-H, trans-[ReCl(N2)(PTA-H)n(PTA)(4-n)]n+ (n = 0-4), are prepared, shown to be soluble and stable in water, interconvertible by stepwise protonation/deprotonation and to form, upon N2 loss, the corresponding penta coordinate compounds. Dinitrogen displacement by CO affords trans [ReCl(CO)(PTA)4]. PMID- 18399234 TI - Homoleptic coordination of arsenine C5H5As to molybdenum(0) and tungsten(0): further observations on the eta1,eta6-dilemma. AB - By means of metal-atom ligand-vapor cocondensation (CC) hexa(eta1 arsenine)molybdenum (11) and hexa(eta1-arsenine)tungsten (12) have been prepared; in the molybdenum case, the sandwich complex bis(eta6-arsenine)molybdenum (10) has been isolated as the primary product. The structure of 12 follows from a single-crystal X-ray diffraction study. Based on the results of CC synthesis, chromium binds to arsenine in the eta6-mode exclusively, molybdenum features both options eta1 and eta6, and for tungsten only eta1 coordination is observed. PMID- 18399235 TI - The gas-phase structure of 1-selena-closo-dodecaborane(11), 1-SeB11H11, determined by the concerted use of electron diffraction and computational methods. AB - The molecular structure of 1-selena-closo-dodecaborane(11), 1-SeB11H11, has been determined by the concerted use of quantum chemical calculations and gas-phase electron diffraction. The structure has C(5v) symmetry and is distorted from a regular icosahedron mainly through the expansion of the pentagon of boron atoms adjacent to selenium, with r(a3,1)(B-B) = 192.2(2) pm. The Se-B bond length is extremely well determined [r(a3,1) (Se-B) = 212.9(2) pm] and this is reflected by a pronounced peak in the radial-distribution curve. The accuracy of the experimental structure, as well as that calculated at the MP2/962(d) level, has been gauged by comparison of the 11B chemical shifts (calculated at two different gauge-including atomic orbitals (GIAO) levels) with experimental NMR values. The inclusion of electron correlation in the magnetic property calculations (GIAO MP2) gave superior results to those carried out using GIAO-Hartree-Fock. PMID- 18399236 TI - The coordination chemistry of perfluorovinyl substituted phosphine ligands, a crystallographic and spectroscopic study. Co-crystallisation of both cis- and trans-isomers of [PtCl2{P(i)pr2(CF=CF2)}2] within the same unit cell. AB - The coordination chemistry of the perfluorovinyl phosphines PEt2(CF=CF2), P(i)Pr2(CF=CF2), PCy,(CF=CF2) and PPh(CF=CF2)2 to rhodium(I), palladium(II), and platinum(II) centres has been investigated. The electronic properties of the ligands are estimated based on v(CO) and 1J(Rh-P) values. X-Ray diffraction data for the square-planar Pd(II) and Pt(II) perfluorovinyl-phosphine containing complexes allow estimates of the steric demand for the series of ligands PPh2(CF=CF2), PEt2(CF=CF2), P(i)Pr2(CF=CF2), PCy2(CF=CF2) and PPh(CF=CF2)2 to be determined. The (CF=CF2) fragment is found to be more electron withdrawing than (C6F5) yet sterically less demanding. These ligands therefore provide a range of electron-neutral to phosphite-like electronic properties combined with a range of steric demands. This study also reveals that short intramolecular interactions from the metal centre to the beta-fluorine atom cis to phosphorus of the CF=CF2 groups are observed in all-trans square planar complexes of the ligands. Unusually, the complex [PtCl2{P(i)Pr2(CF=CF2)}2] crystallises with both cis- and trans-isomers present in the unit cell. It appears that co-crystallisation of both isomers occurs in order to maximise fluorous regions in the crystal packing, and the extended structure displays short fluorine-fluorine contacts. The generation of mixed geometries seems to be a phenomenon of crystallisation, as solution phase NMR studies reveal the presence of only the trans-isomer. PMID- 18399237 TI - Organic-inorganic hybrids assembled by bis(undecatungstophosphate) lanthanates and dinuclear copper(II)-oxalate complexes. AB - A series of organic-inorganic hybrid compounds, K2H7[{Ln(PW11O39)2}{Cu2(bpy)2(mu ox)}].xH2O (Ln = La, x approximately = 18 (1); Ln = Pr, x approximately = 18(2); Ln = Eu, x approximately = 16(3); Ln = Gd, x approximately 22(4); Ln = Yb, x approximately = 19 (5); bpy = 2,2'-bipyridine and ox = oxalate), have been isolated by the conventional solution method. Single-crystal X-ray diffraction studies reveal that compounds 1-5 are isomorphic and consist of one-dimensional chains, which are constructed by alternating bis(undecatungstophosphate) lanthanates [Ln(PW11O39)2](11-) and dinuclear copper(II)-oxalate complexes [Cu2(bpy)2(mu-ox)]2+.pi-pi interactions of the bpy ligands from adjacent chains lead to their three-dimensional structures. An analogue of potassium K2H9[{K(PW11O39)2}{Cu2(bpy)2(mu-ox)}1].approximately 20.5H2O(6) has also been obtained. The syntheses and structures of these compounds are reported here. Magnetic properties of 1, 2 and 3 are discussed as well. Attempts to crystallize similar compounds containing Co(II) and Ni(II) were unsuccessful. PMID- 18399239 TI - Steric control of the nuclearity of metallamacrocycles: formation of a hexanuclear gallium metalladiazamacrocycle and a hexadecanuclear manganese metalladiazamacrocycle. AB - An S6-symmetric hexanuclear gallium metalladiazamacrocycle, [Ga(III)(6)L2(6)S6] with a -(lamda delta)(lamda delta)-chiral sequence and an S-symmetric hexadecanuclear manganese metalladiazamacrocycle, [Mn(III)(16)L2(16)S16] with a (lamda lamda delta delta)(lamda lamda delta delta)-chiral sequence were prepared using the same N2-trans-cinnamoyl-2-hydroxy-3-naphthoylhydrazide (H3L2) as a bridging pentadentate ligand between the metal centers for the formation of a macrocyclic system. PMID- 18399238 TI - Interpretation of the multiple vanadium-oxygen bonds in the central VO(eta2-O2)+ group. Synthesis, structure, supramolecular interactions and DFT studies for complexes with 2,2'-bipyridine, 1,10-phenanthroline, pyrazinato(1-) and pyrazinamide ligands. AB - Neutral peroxovanadium(v) complexes, [VO(O2)(pca)(bpy)] (1), [VO(O2)(pca)(phen)] (2) and [VO(O2)(pic)(pcaa)(H2O)].H2O(3), were synthesized [2,2'-bipyridine (bpy), 1,10-phenanthroline (phen), pyrazinecarboxamide (pcaa), 2-pyrazinecarboxylic (Hpca) and picolinic (Hpic) acids]. Their X-ray single crystal analysis revealed a distorted pentagonal bipyramidal geometry in all complex molecules. The four "free" coordination sites of the vanadium atoms of the VO(eta2-O2)+ moieties in 1 and 2 are occupied by the donor atoms of two bidentate heteroligands. The supramolecular structures of 1 and 2 are exclusively constructed by intermolecular C--H(ar)...O hydrogen bonds [dH(H...O): 2.292-2.708 A (1), and 2.260-2.720 A (2)]. In addition, the structures are stabilized by parallel off set pi-pi interactions between the bpy rings resp. non-parallel off-set interactions between the phen rings [centroid distances: 3.7000(1) A (1), 3.9781(2) and 3.6757(2) A (2)]. In the molecular structure of 3, pcaa is coordinated in an equatorial position of the bipyramid via the nitrogen atom of the pyrazine ring, while the aqua ligand is in the apical position. The disordered crystal water molecules are located in 1D channels oriented along the a axis. The intermolecular C-H(ar)...O hydrogen bonds in 3 were found within the dH(H..O) range 2.409-2.669 A. The pic ligands are off-set pi-pi stacked, with centroid distances: 3.6725(3) and 3.8323(3) A. The DFT orbital calculations and NBO analysis for the VO(eta2-O2)+ group gave evidence for a triple V[triple bond]O bond, and showed that the observed cis arrangement of the oxo and peroxo ligands results from the direct interaction between them. Experimental and calculated UV-Vis and IR spectral data are presented. PMID- 18399240 TI - Monohydroxamic acids and bridging dihydroxamic acids as chelators to ruthenium(III) and as nitric oxide donors: syntheses, speciation studies and nitric oxide releasing investigations. AB - The synthesis and spectroscopic characterisation of novel mononuclear Ru(III)(edta)(hydroxamato) complexes of general formula [Ru(H2edta)(monoha)] (where monoha = 3- or 4-NH2, 2-, 3- or 4-C1 and 3-Me-phenylhydroxamato), as well as the first example of a Ru(III)-N-aryl aromatic hydroxamate, [Ru(H2edta)(N-Me bha)].H2O (N-Me-bha = N-methylbenzohydroxamato) are reported. Three dinuclear Ru(III) complexes with bridging dihydroxamato ligands of general formula [{Ru(H2edta)}2(mu-diha)] where diha = 2,6-pyridinedihydroxamato and 1,3- or 1,4 benzodihydroxamato, the first of their kind with Ru(III), are also described. The speciation of all of these systems (with the exception of the Ru-1,4 benzodihydroxamic acid and Ru-N-methylbenzohydroxamic systems) in aqueous solution was investigated. We previously proposed that nitrosyl abstraction from hydroxamic acids by Ru(III) involves initial formation of Ru(III)-hydroxamates. Yet, until now, no data on the rate of nitric oxide (NO) release from hydroxamic acids has been published. We now describe a UV-VIS spectroscopic study, where we monitored the decrease in the ligand-to-metal charge-transfer band of a series of Ru(III)-monohydroxamates with time, with a view to gaining an insight into the NO releasing properties of hydroxamic acids. PMID- 18399241 TI - Mixed ligand ruthenium(II) complexes of bis(pyrid-2-yl)-/bis(benzimidazol-2-yl) dithioether and diimines: study of non-covalent DNA binding and cytotoxicity. AB - A series of mixed ligand ruthenium(II) complexes [Ru(pdto)(diimine)](ClO4)2/(PF6)2 1-3 and [Ru(bbdo)(diimine)](ClO4), 4-6, where pdto is 1,8-bis(pyrid-2-yl)-3,6-dithiooctane, bbdo is 1,8-bis(benzimidazol-2-yl) 3,6-dithiooctane and diimine is 1,10-phenanthroline (phen), dipyrido-[3,2-d:2',3' f]-quinoxaline (dpq) and dipyrido[3,2-a:2',3'-c]phenazine (dppz), have been isolated and characterised by analytical and spectral methods. The complexes [Ru(pdto)(phen)](PF6)2 la, [Ru(pdto)(dpq)(Cl](PF6) 2a, [Ru(bbdo)(phen)](PF6)2 4a and [Ru(bbdo)(dpq)](ClO4)2 5 have been structurally characterized and their coordination geometries around ruthenium(II) are described as distorted octahedral. In la, 4a and 5 the two thioether sulfur and two py/bzim nitrogen atoms of the tetradentate pdto/bbdo ligand are folded around Ru(II) to give predominantly a "cis-alpha" configuration. (I)H NMR spectral data of the complexes support this configuration in solution. In [Ru(pdto)(dpq)Cl](PF6) 2a with a distorted octahedral coordination geometry, one of the two py nitrogens of pdto is not coordinated. The DNA binding constants (Kb: 2, 2.00 +/- 0.02 x 10(4) M(-1), s = 1.0; 3, 3.00 +/- 0.01 x 10(6) M(-1), s = 1.3) determined by absorption spectral titrations of the complexes with CT DNA reveal that 3 interacts with DNA more tightly than 2 through partial intercalation of the extended planar ring of coordinated dppz with the DNA base stack. The DNA binding affinities of the complexes increase with increase in the number of planar aromatic rings in the co ligand, and on replacing both the py moieties in pdto complexes (1-3) by bzim moieties to give bbdo complexes (4-6). Upon interaction with CT DNA the complexes 1, 2, 5 and 6 show a decrease in anodic current in the cyclic voltammograms. On the other hand, interestingly, 3 and 4 show an increase in anodic current suggesting their involvement in electrocatalytic guanine oxidation. Interestingly, of all the complexes, only 6 alters the superhelicity of DNA upon binding with supercoiled pBR322 DNA. The cytotoxicities of the dppz complexes 3 and 6, which avidly bind to DNA, have been examined by screening them against cell lines of different cancer origins. It is noteworthy that 6 exhibits selectivity with higher cytotoxicity against the melanoma cancer cell line (A375) than other cell lines, potency approximately twice that of cisplatin and toxicity to normal cells 3 and 90 times less than cisplatin and adriamycin respectively. PMID- 18399242 TI - H-atom abstraction reaction for organic substrates via mononuclear copper(II) superoxo species as a model for DbetaM and PHM. AB - Hydrogen atom abstraction reactions have been implicated in oxygenation reactions catalyzed by copper monooxygenases such as peptidylglycine alpha-hydroxylating monooxygenase (PHM) and dopamine beta-monooxygenase (DbetaM). We have investigated mononuclear copper(I) and copper(II) complexes with bis[(6 neopentylamino-2-pyridyl)methyl][(2-pyridyl)methyl]amine (BNPA) as functional models for these enzymes. The reaction of [Cu(II)(bnpa)]2+ with H2O2, affords a quasi-stable mononuclear copper(II)-hydroperoxo complex, [Cu(II)(bnpa)(OOH)]+ (4) which is stabilized by hydrophobic interactions and hydrogen bonds in the vicinity of the copper(II) ion. On the other hand, the reaction of [Cu(I)(bnpa)]+ (1) with O2 generates a trans-mu-1,2-peroxo dicopper(II) complex [Cu(II)2(bnpa)2(O2(2-]2+ (2). Interestingly, the same reactions carried out in the presence of exogenous substrates such as TEMPO-H, produce a mononuclear copper(II)-hydroperoxo complex 4. Under these conditions, the H-atom abstraction reaction proceeds via the mononuclear copper(II)-superoxo intermediate [Cu(II)(bnpa)(O2-)]+ (3), as confirmed from indirect observations using a spin trap reagent. Reactions with several substrates having different bond dissociation energies (BDE) indicate that, under our experimental conditions the H-atom abstraction reaction proceeds for substrates with a weak X-H bond (BDE < 72.6 kcal mol(-1)). These investigations indicate that the copper(II)-hydroperoxo complex is a useful tool for elucidation of H-atom abstraction reaction mechanisms for exogenous substrates. The useful functionality of the complex has been achieved via careful control of experimental conditions and the choice of appropriate ligands for the complex. PMID- 18399243 TI - Bayes Window (5): analogue Bayes? PMID- 18399244 TI - Speed constancy and the perception of distance. AB - The distance-calibration hypothesis states that retinal velocity is scaled by using distance cues, and judged velocity remains unchanged when distance is changed. The relational hypothesis states that judged velocity depends on retinal velocities, and is proportional to judged distance. These hypotheses were compared in three experiments where the movements of the standard stimulus and the comparison stimulus were manipulated by the ratio of the angular velocity of the comparison stimulus to the angular velocity of the standard stimulus. The presentation conditions of the standard stimulus and the comparison stimulus, and the colour cues of the two stimuli were also manipulated in order to change the strength of the cues available to the observers. The results indicate that judged velocities and the relationship of judged distance and velocity depend on the strength of the cues. When cues are strong, the distance-calibration hypothesis adequately explains speed constancy. When cues are weak, judged velocity and the relationship between judged distance and velocity are consistent with the prediction of the relational hypothesis. The perceived speed of a stimulus depends not only on the physical speed of the stimulus but also on non-motion cues, some of which are distance cues involved in depth perception. PMID- 18399245 TI - Accelerating self-motion displays produce more compelling vection in depth. AB - We examined the vection in depth induced when simulated random self-accelerations (jitter) and periodic self-accelerations (oscillation) were added to radial expanding optic flow (simulating constant-velocity forward self-motion). Contrary to the predictions of sensory-conflict theory frontal-plane jitter and oscillation were both found to significantly decrease the onsets and increase the speeds of vection in depth. Depth jitter and oscillation had lesser, but still significant, effects on the speed of vection in depth. A control experiment demonstrated that adding global perspective motion which simulated a constant velocity frontal-plane self-motion had no significant effect on vection in depth induced by the radial component of the optic flow. These results are incompatible with the notion that constant-velocity displays produce optimal vection. Rather, they indicate that displays simulating self-acceleration can often produce more compelling experiences of self-motion in depth. PMID- 18399246 TI - Monitoring eye movements to investigate the picture superiority effect in spatial memory. AB - Spatial memory is usually better for iconic than for verbal material. Our aim was to assess whether such effect is related to the way iconic and verbal targets are viewed when people have to memorize their locations. Eye movements were recorded while participants memorized the locations of images or words. Images received fewer, but longer, gazes than words. Longer gazes on images might reflect greater attention devoted to images due to their higher sensorial distinctiveness and/or generation with images of an additional phonological code beyond the visual code immediately available. We found that words were scanned mainly from left to right while a more heterogeneous scanning strategy characterized encoding of images. This suggests that iconic configurations tend to be maintained as global integrated representations in which all the item/location pairs are simultaneously present whilst verbal configurations are maintained through more sequential processes. PMID- 18399247 TI - The effect of temporal and spatial frequency on phantom-contour detection. AB - Phantom contours are a visual illusion that can define regions with distinctive boundaries when no real surrounding edges exist. Spatial-frequency sensitivity is known to vary reliably across the visual-processing pathways, as does temporal frequency sensitivity. Given that the effect of temporal frequency on phantom contour detection has been previously established, and that the relationship between spatial frequency and temporal frequency is known, two experiments were designed to measure the highest level of spatial frequency that would still allow reliable pattern detection at different temporal frequencies by using the phantom contour paradigm. The results revealed that phantom-contour detection is impaired when the stimulus has a high spatial-frequency content and that phantom-contour perception is supported by low spatial frequencies. PMID- 18399248 TI - Independent processing of form, colour, and texture in object perception. AB - Most investigations of object recognition have focused on the form rather than the material properties of objects. Nevertheless, knowledge of the material properties of an object (via its surface cues) can provide important information about that object's identity. In this study, we used Garner's speeded classification task to explore whether or not the processing of form and the processing of surface properties are independent. In experiment 1, participants made length and width classifications in an initial form task. Participants were unable to ignore length while making width classifications, and were unable to ignore width while making length classifications. This suggests that the perception of length and the perception of width share common processing resources. In a subsequent task, we examined possible interactions between the processing of form and the processing of surface properties. In contrast to the findings with the form task, participants were able to ignore form while making surface-property classifications, and to ignore surface properties while making form classifications. This suggests that the form of objects and their surface properties are processed independently. In experiment 2, we went on to show that the two prominent surface-property dimensions of colour and texture can also be processed independently. In other words, participants were able to ignore colour while making texture classifications, and vice versa. Finally, in experiment 3, we examined the possibility that the stimuli and required responses that we used in experiment 2 were too categorical and thus not optimal for assessing whether or not colour and texture share common processing resources. Using a different stimulus set, participants were again able to ignore colour while making texture classifications, and vice versa. Taken together, these results provided convincing evidence that the separate ventral-stream brain regions identified for form, texture, and colour in a recent neuroimaging study (Cant and Goodale, 2007 Cerebral Cortex 17 713-731) can indeed function independently. PMID- 18399249 TI - The intrinsic constraint model for stereo-motion integration. AB - How the visual system integrates the information provided by several depth cues is central for vision research. Here, we present a model for how the human visual system combines disparity and velocity information. The model provides a depth interpretation to a subspace defined by the covariation of the two signals. We show that human performance is consistent with the predictions of the model, and compare them with those of another theoretical approach, the modified weak-fusion model. We discuss the validity of each approach as a model for human perception of 3-D shape from multiple cues to depth. PMID- 18399250 TI - Event-related potentials, reaction time, and response selection of skilled and less-skilled cricket batsmen. AB - The differences in P300 latency, P300 amplitude, response selection, and reaction time between skilled and less-skilled cricket batsmen have been investigated. Eight skilled and ten less-skilled right-handed batsmen each viewed 100 in-swing, 100 out-swing, and 40 slower deliveries displayed in random sequence from projected video footage whilst their responses and electroencephalograms were recorded. Logistic regression was used to derive a discriminative function for the P300 data. This was done to determine whether the skilled batsmen differed from the less-skilled batsmen on the basis of pooled P300 amplitude and latency data. All the batsmen were correctly characterised as being skilled or less skilled. Logistic regression equations with reaction time and correctness of response data indicated that behavioural data do not correctly classify skilled performance. It is suggested that skilled cricket batsmen have a superior perceptual decision-making ability compared with less-skilled cricket batsmen, as measured by P300 latency and amplitude. This appears to be the first study showing a link between skill and cerebral cortical activity during a perceptual cricket batting task and it could pave the way for future studies on mental processing in cricket batsmen. PMID- 18399251 TI - Is pictorial perception robust? The effect of the observer vantage point on the perceived depth structure of linear-perspective images. AB - Every image of a scene produced in accord with the rules of linear perspective has an associated projection centre. Only if observed from that position does the image provide the stimulus which is equivalent to the one provided by the original scene. According to the perspective-transformation hypothesis, observing the image from other vantage points should result in specific transformations of the structure of the conveyed scene, whereas according to the vantage-point compensation hypothesis it should have little effect. Geometrical analyses illustrating the transformation theory are presented. An experiment is reported to confront the two theories. The results provide little support for the compensation theory and are generally in accord with the transformation theory, but also show systematic deviations from it, possibly due to cue conflict and asymmetry of visual angles. PMID- 18399252 TI - Computational models of facial attractiveness judgments. AB - We designed two computational models to replicate human facial attractiveness ratings. The primary model used partial least squares (PLS) to identify image factors associated with facial attractiveness from facial images and attractiveness ratings of those images. For comparison we also made a model similar to previous models of facial attractiveness, in that it used manually derived measurements between features as inputs, though we took the additional step of dimensionality reduction via principal component analysis (PCA) and weighting of PCA dimensions via a perceptron. Strikingly, both models produced estimates of facial attractiveness that were indistinguishable from human ratings. Because PLS extracts a small number of image factors from the facial images that covary with attractiveness ratings of the images, it is possible to determine the information used by the model. The image factors that the model discovered correspond to two of the main contemporary hypotheses of averageness judgments: facial attractiveness and sexual dimorphism. In contrast, facial symmetry was not important to the model, and an explicit feature-based measurement of symmetry was not correlated with human judgments of facial attractiveness. This provides novel evidence for the importance of averageness and sexual dimorphism, but not symmetry, in human judgments of facial attractiveness. PMID- 18399253 TI - Investigating the effects of inversion on configural processing with an audiovisual temporal-order judgment task. AB - Research has shown that inversion is more detrimental to the perception of faces than to the perception of other types of visual stimuli. Inverting a face results in an impairment of configural information processing that leads to slowed early face processing and reduced accuracy when performance is tested in face recognition tasks. We investigated the effects of inverting speech and non-speech stimuli on audiovisual temporal perception. Upright and inverted audiovisual video clips of a person uttering syllables (experiments 1 and 2), playing musical notes on a piano (experiment 3), or a rhesus monkey producing vocalisations (experiment 4) were presented. Participants made unspeeded temporal-order judgments regarding which modality stream (auditory or visual) appeared to have been presented first. Inverting the visual stream did not have any effect on the sensitivity of temporal discrimination responses in any of the four experiments, thus implying that audiovisual temporal integration is resilient to the effects of orientation in the picture plane. By contrast, the point of subjective simultaneity differed significantly as a function of orientation only for the audiovisual speech stimuli but not for the non-speech stimuli or monkey calls. That is, smaller auditory leads were required for the inverted than for the upright-visual speech stimuli. These results are consistent with the longer processing latencies reported previously when human faces are inverted and demonstrates that the temporal perception of dynamic audiovisual speech can be modulated by changes in the physical properties of the visual speech (ie by changes in orientation). PMID- 18399254 TI - 17,000 years of depicting the junction of two smooth shapes. AB - Competent realistic drawings preserve viewpoint-invariant shape characteristics of simple parts, such that a contour in the object that is straight or curved, for example, is depicted that way in the drawing. A more subtle invariant--a V shaped singularity of the occluding boundary, containing a T-junction and a contour termination--is produced at the junction between articulated smooth surfaces, as with the leg joining the body of a horse. 45% of the drawings made in 2007 by individuals with only minimal art education correctly depicted such junctions, a proportion that is not reliably different from the incidence (42%) of correct depictions in a large sample of cave art made 17000 years ago. Whether a person did or did not include the invariant in their drawing, all agreed that it made for a better depiction. PMID- 18399255 TI - [16-MDCT angiography coronary artery in the emergency department for patients with acute coronary syndrome (NSTEMI-UA)]. AB - AIM: The diagnosis of acute coronary syndrome (ACS), non-ST-elevation myocardial infarction and unstable angina in the emergency department (ED) remains a challenge. The aim of our study was to investigate quality and the diagnostic accuracy of 16-MDCT coronary angiography, detecting coronary artery lesions in patients with suspected ACS presented in ED. MATERIALS AND METHODS: We studied with 16-MDCT (Sensation 16, Siemens, Forchleim, Germania) and coronary angiography 37 patients with the following inclusion criteria: chest pain compatible with myocardial ischemia, normal or no-diagnostic ECG changes and initial concentrations of serum troponin-I < or =1 ng/ml. The 16-MDCT was performed with ECG-gated technique after the intravenous administration of 90-100 ml of iodinated contrast material followed by a saline bolus. The scan parameters were: 120 kV, 650-720 mAs, 16 x 0.75mm collimation, 0.42s rotation time, 3 mm (pitch 0.25) feed/rot, B30f kernel. We evaluated for each patient: image quality and different artefacts, plaques identification and characterization. RESULTS: The evaluation of the image quality was based on a total of 453 segments, of which 415 segments (92.2%) were considered to have diagnostic image quality. MDCT correctly detected 15 patients with at least 1 stenosis >50% and correctly ruled out significant coronary artery disease in 19 patients with 1 FP and 2 FN: sensitivity 88%, specificity 95%, PPV 94%, NPV 90%. The plaques were hard in 6 cases, mixt in 16 cases and soft in 14 cases, respectively. CONCLUSIONS: Our results point-out that 16-MDCT in ED has the real ability to detect and rule out significant coronary stenoses in patients with ACS. PMID- 18399256 TI - [Treatment of mild malnutrition and reduction of morbidity in major abdominal surgery: randomized trial on 153 patients]. AB - OBJECTIVES: Severe malnutrition (defined as weight loss more than 10% in a period of six months) is considered an important risk factor in major abdominal surgery, because of a higher post-operative mortality and morbidity. The aim of our study is to assess the role of mild malnutrition (weight loss low than 10% in a period of six months) as a risk factor in major abdominal surgery and to evaluate the efficacy of therapy in order to improve outcomes in terms of in-hospital mortality, length of hospital stay and post-operative complications. Moreover, we evaluated serum albuminemia and lymphocyte count, important nutritional index, as predictive risk factors. MATERIALS AND METHODS: We performed a randomized prospective trial, and admitted in our institution 153 adult patients, 43 with mild malnutrition and 110 without. The malnourished patients were randomized in two groups: the first one received oral immunonutrition (Impact Oral) for 7-10 days before surgery (22 pz), the second one received no nutritional support. RESULTS: We observed a higher number of complications in the non-treated malnourished patients (57%) versus both the treated malnourished patients (13.6%) and the normal group (19%) (p<0.001). Increased morbidity was observed in patients with serum albuminemia <2.8 gr/dl (69.2%) and with lymphocyte count <1.500 mm3 (57%). CONCLUSIONS: Nutritional enriched support demonstrated his efficacy in reducing morbidity, and length of hospital stay. Pre-operative oral immunonutrition might be suggested and established in all the patients with mild malnutrition that will be operated on major abdominal surgery. PMID- 18399257 TI - [Sedation with propofol in endoscopic retrograde cholangiopancreatography: personal experience]. AB - AIM: Adequate sedation is fundamental for the execution of the endoscopic retrograde cholangiopancreatography (ERCP). Propofol is widely used for gastrointestinal endoscopy because of its rapid recovery profile. The aim of this study was to determine, retrospectively, whether the administration of propofol was safe in patients undergoing ERCP, both diagnostic and therapeutic. MATERIALS AND METHODS: In our GI Unit, from 1st February 2006 to 23 November 2006, we performed 100 ERCP. All the patients were sedated by using midazolam e.v., as pre anaesthetic agent, and propofol e.v. During the procedure, vital signs were continuously monitored (oxygen saturation, blood pressure, heart rate). Patients were also divided into two groups: less than 80 years of age (group I) and 80 years of age and older (group II). Cardiorespiratory complications were recorded. RESULTS: Patients were 51 females and 49 males, with a median age of 74 years (range: 23-94 years). Group I was composed by 72 patients (35 F, 37 M) and Group II by 28 patients (16 F, 12 M). There were no episodes of hemodynamic instability or airway obstruction. New ECG changes (1 ischemia, 3 arrhythmias) and 1 significant oxygen desaturation episode (SpO2<90%) occurred in 5% of procedures. If we considered the two groups, the rates of cardiopulmonary complications were 4.1% and 7.1%, respectively in group I and in group II. CONCLUSIONS: Propofol seems to be safe and effective sedation for ERCP, with a low complication rate, also in patients aged 80 years or older. PMID- 18399259 TI - [Bioethics and pharmacoeconomics: a difficult balance]. AB - While economic resources continue to decrease, there is a growing demand for health treatment. This faces health workers with an ethical dilemma. They are caught between, on the one hand, their responsibility to the individual patient and, on the other hand, the obvious need to make new treatments available to the largest number of patients. This clearly highlights the need for a regulated system of allocation of resources, whose rules must be agreed to by all operators in the field. PMID- 18399258 TI - [The surgical treatment of renal cell carcinoma metastases to the vertebrae: intra-operative anesthesia related problems]. AB - AIMS: Analysis of the intra-operative anesthesia related problems were compared during intra-lesional or extra-lesional surgeries for patients with metastatic renal cell carcinoma to the spine. MATERIALS AND METHODS: We retrospectively collected and analyzed the hemodynamic data on 30 patients who had undergone surgical intervention. Twenty patients had an intra-lesional excision of the metastatic lesion in the vertebra, whereas for the rest, an en-bloc wide or marginal margin resection was carried out. RESULTS: The surgical time for the en bloc resections was much longer, and thus also the exposure to the anesthesia, but lesser variability within the data on blood pressure and pulse as compared with intra-lesional curettage. There was also more blood loss with the intra lesional surgeries. CONCLUSIONS: En-bloc resections of the vertebral tumors not only give better oncological local control, but also have less compromising effects on the patient, especially with respect to the intra-operative hemodynamics. In selected cases, en-bloc resections present a good option of treatment in vertebral metastases, and especially in highly vascular tumors such as the renal cell carcinoma metastases. PMID- 18399260 TI - Concurrent tonsillitis and pneumonia due to S. pyogenes. AB - Tonsillitis is a common disease especially of children and young adults. The diagnosis of tonsillitis generally requires the consideration of Group A beta hemolytic streptococci infection. Pneumonia is an inflammation of the alveoli and terminal airspaces in response to invasion by an infectious agent that is introduced into the lungs through hematogenous spread or inhalation. We present a case of a concurrent acute tonsillitis and pneumonia due to S. pyogenes. To our knowledge, a similar case has not been previously described in the literature. PMID- 18399261 TI - Uric acid and renal function in multiple sclerosis. AB - OBJECTIVE: The purpose of this study was to examine the circadian distribution of creatinine and uric acid clearances in subjects with Multiple Sclerosis. MATERIALS AND METHODS: Eleven subjects with MS, 6 women (48+/-7y) and 5 men (58+/ 5y) volunteered for this circadian study. Thirteen healthy females (39+/-11y) served as controls. Data of seven healthy male controls (64+/-8 y) were extracted from a similar circadian study conducted previously. Each MS patient, and each male control had blood samples drawn around the clock, at 3h intervals (8/24h), and each collected urines over 3h periods (8/24h). Each female control contributed only one blood sample and one complete 24h urine collection. Blood and urine samples were analyzed for a number of relevant analytes: ELAM, IL-6, NO, insulin, ACTH, aldosterone, cortisol, electrolytes, lymphocytes, monocytes including creatinine and uric acid clearances. Those were standardized to an average body surface area of 1.73 m2. RESULTS: The relevant analytes demonstrated increased synthesis of insulin, IL-6, ELAM, monocytes, and reduced concentrations of serum NO. The creatinine clearances were significantly lower in MS females than in female controls, 63+/-22 vs.108+/-18 ml/min. They were also lower than those of MS males and male controls, 107.8+/-17, 97.5+/-8.2 ml/min. Uric acid clearances in MS females were also lower 6.9+/-2.4 vs. 10.5+/-4.4 ml/min. The uric acid clearance in MS males was higher than in male controls, 7.0+/-4.5 vs. 4.0+/-1.0 ml/min. CONCLUSIONS: The alterations in selected relevant analytes and the reduced creatinine and uric acid clearances in females but not in males, suggest a renal dysfunction in MS females. These observations may contribute to understanding better the mechanism of renal dysfunction in female patients and perhaps this may be an additional factor contributing to greater frequency of MS in females than in male subjects. PMID- 18399262 TI - Management of chronic hepatitis in drug addicts: a systematic review. AB - Injection drug users constitute the largest group of person at high risk for acquiring chronic hepatitis C, B and Delta. In particular viral, host and environmental factors all seem to favour rapid spread of these infections among drugs addicts. Host factors include behaviours that expose individuals to hepatitis virus such as the shared use of drug preparation, injection equipment and not protected sexual relationship with other drugs users. While in some clinical studies adherence to treatment regimens was often poor and to treat chronic hepatitis in injection drug users was stated as futile, in other controlled clinical studies adherence and sustained biological response to antiviral treatment was slightly lower or similar to that reported in other groups of patients. In this review we describe the epidemiology, diagnosis and treatment of chronic hepatitis C, B and Delta in intravenous drug users. PMID- 18399263 TI - Vascular injury, hypertension and coronary artery disease in human immunodeficiency virus infection. AB - The atherogenic effects of some highly active antiretroviral therapy (HAART) regimens, especially those including protease inhibitors (PI), may synergistically promote the acceleration of cardiovascular disease and increase the risk of death from cardiovascular events even in young HIV-infected people. Along with the endothelial dysfunction associated with visceral fat accumulation and related metabolic alterations of HIV-lipodystrophy syndrome (eg, insulin resistance), vascular injury has been associated with HIV-1 infection itself, with an autoimmune reaction to viral infection (vasculitis) and with a direct action of drugs included in HAART regimens. Clinical studies suggest that HIV infected patients under PI-including HAART and with preexisting cardiovascular risk factors, should be considered at risk for developing coronary artery disease, and that this risk increases with the time of exposure to HAART. A careful cardiovascular screening and monitoring of HIV-infected patients receiving HAART is needed according to the most recent clinical guidelines. PMID- 18399264 TI - Stakeholder engagement and public policy evaluation: factors contributing to the development and implementation of a regional network for geriatric care. AB - The development and implementation of a regional network that provides universally accessible and consistent services to the frail elderly living in Southwestern Ontario is described. Through continuous stakeholder engagement, clear network goals were identified and operationalized. Stakeholder commitment to the integration of expertise and specialized services, to evidence-based public policy and to iterative evaluation cycles were key to network success. PMID- 18399265 TI - Enhancing service delivery capacity through knowledge exchange: the Seniors Health Research Transfer Network. AB - The Seniors Health Research Transfer Network (SHRTN) was launched in 2005 in Ontario to improve the quality of health care provided to seniors by facilitating knowledge exchange opportunities for formal and informal caregivers, researchers and policymakers. This article describes the conceptual basis and development of SHRTN, as well as achievements, challenges and lessons learned during its first year of operation, which ended in March 2006. We begin by discussing knowledge exchange networks and their conceptual basis. We then offer a brief history of SHRTN, tracing its origins to both a broad interprofessional interest in creating and sharing knowledge within and across organizations, and also to the efforts of a small group of early champions. After this, we describe the main events, achievements and surprises of SHRTN's first year. Experience with SHRTN has highlighted the importance of careful attention to governance issues in the organization of knowledge exchange networks, and the challenge of balancing management control with broad participation and flexibility. Collaboration can yield synergy and innovation, but requires commitment from participants. The SHRTN experience has demonstrated that planning and coordinating a provincial network that engages diverse stakeholders is a logistical challenge that requires dedicated infrastructure and funding support. PMID- 18399266 TI - Disease management in Canada: surmounting barriers to adoption. AB - Disease Management (DM) programs are used to optimize economic outcomes and improve patient outcomes. Despite this, relative to the United States, Canadian health care organizations have been slow to adopt them. The objective of this article is to examine the concept of DM programs, the existing evidence to support their use and the barriers to their adoption in Canada. Several solutions aimed at overcoming the barriers to DM in Canada are proposed. PMID- 18399267 TI - Public-private partnerships: a Canadian hospital's perspective. AB - The McGill University Health Centre (MUHC) is undergoing a $1.579 billion redevelopment project, placing it amongst the largest hospital redevelopment projects in the world. As the Quebec government and the MUHC were considering the Public-Private Partnership (PPP) procurement alternative, the MUHC embarked upon an investigation of various jurisdictions' experiences with PPPs. The studies concluded that there are various frameworks available and that the specific characteristics of a project should be considered when determining which model to use. PMID- 18399268 TI - How can frontline managers demonstrate leadership in enabling interprofessional practice? AB - Interprofessional practice has been proposed to address many of the current health system challenges. Little attention has been paid to the role of frontline managers in supporting effective interprofessional practice. Qualitative interviews suggest that frontline managers can demonstrate leadership in enabling interprofessional practice by creating an organizational culture for interprofessional practice, eliminating barriers to collaboration, and acting as role models and facilitators. New management approaches may be needed to meet these demands. PMID- 18399269 TI - Individual and typological behavioral characteristics of CBA/CaLac mice. AB - Individual characteristics of higher nervous activity were studied in CBA/CaLac mice. The animals were divided into groups by the parameters of drinking response conditioning in a complex spatial environment. As distinct from bad learners, good learners exhibited higher orientation and exploratory activity in the open field, rapid adaptation, and low ability to retain the responses. Changes in conditioned reflex activity during neuroses were more pronounced in good learners. PMID- 18399270 TI - Cholinergic and GABAergic innervation regulate activity of electrogenic ionic pump in earthworm somatic muscle cells. AB - Carbacholine reduced, while baclofen and norepinephrine increased resting membrane potential in earthworm somatic muscle cells. In the presence of carbacholine, neither norepinephrine, nor baclofen hyperpolarized the membrane. Ouabain decreased resting potential and abolished the effects of carbacholine, norepinephrine, and baclofen on membrane potential. It was hypothesized that carbacholine directly inhibited the ouabain-sensitive component of Na+/K+ pump and abolished the activating effect of norepinephrine and baclofen. PMID- 18399271 TI - Parasympathetic cardiac effects in sympathectomized rats. AB - In adult sympathectomized rats the right and left vagus nerves exhibited asymmetric effects on heart rate and strength of cardiac contractions. After vagotomy, the heart rate and stroke volume remained high in sympathectomized rats, but returned to normal in intact animals. The sympathetic nervous system plays a role in postvagotomic tachycardia. Sympathectomized rats were characterized by higher reactivity of cardiac beta-adrenoceptors to propranolol compared to intact animals. PMID- 18399272 TI - M3 cholinergic receptors are involved in postnatal development of cholinergic regulation of cardiac activity in rats. AB - We studied the role of M3 cholinergic receptors in the regulation of cardiac activity in rats during early postnatal ontogeny in vivo. Blockade of M3 cholinergic receptors in 20-week-old animals increased heart rate and decreased blood pressure. In rats aging 8, 6, and 3 weeks, blockade of M3 cholinergic receptors had little effect on R-R interval, but unexpectedly increased it in 1 week-old animals. It can be hypothesized that tonic inhibitory effect of the vagus nerve in adult rats is realized through M3 cholinergic receptors of the heart. The decrease in heart rate during blockade of M3 cholinergic receptors in 1-week-old rats was probably related to specific innervation of the heart in animals of this age. PMID- 18399273 TI - Role of heat shock proteins HSP70 and HSP32 in the protective effect of adaptation of cultured HT22 hippocampal cells to oxidative stress. AB - Preadaptation of cultured HT22 mouse hippocampal neurons to oxidative stress prevented cell damage induced by severe oxidative stress. This protection manifested in a decrease in metabolic disturbances in neurons. Adaptation of neurons to oxidative stress was accompanied by accumulation of HSP32 and HSP70. HSP synthesis inhibitor quercetin abolished the protective effect of adaptation under conditions of oxidative stress. Activation of HSP70 synthesis in neurons is an important mechanism for adaptive protection of cells. PMID- 18399274 TI - Effects of alpha1-acid glycoprotein on hemostasis in experimental septic peritonitis. AB - Pathogenesis of hemostasis disorders in septic peritonitis and the possibility of their correction with acute phase protein (alpha1-acid glycoprotein; two doses of 150 mg/kg) were experimentally studied on outbred albino rats. Platelets count in the peripheral blood and their adhesion to endothelium did not change during peritonitis, while aggregation activity increased due to increased rate and shorter time of aggregation, which was associated with the development of hypercoagulation involving the intrinsic and common coagulation pathways and reduction of antithrombin activity. alpha1-Acid glycoprotein increased platelet count above the normal level, normalized aggregation rate, some blood clotting parameters, and antithrombin activity. Hence, alpha1-acid glycoprotein is a polyfunctional protein modulating all pathogenetic components in the development of blood clotting disorders during septic peritonitis. PMID- 18399275 TI - Early and late hemopoietic precursor cells in patients with chronic myeloproliferative diseases. AB - We evaluated the content of early and late cobblestone area-forming cells, immediate progeny of hemopoietic stem cells, and committed precursor cells in the bone marrow and peripheral blood of patients with chronic myeloproliferative diseases and healthy donors. In patients with essential thrombocythemia, the number of late cobblestone area-forming cells in the peripheral blood decreased, while other parameters did not differ from those in healthy donors. In patients with idiopathic myelofibrosis, we found a decreased number of late and early cobblestone area-forming cells in the bone marrow and late cobblestone area forming cells in the peripheral blood, while the count of early cobblestone area forming cells in the peripheral blood increased. In patients with chronic myeloid leukemia, the number of early cobblestone area-forming cells in the bone marrow decreased, but the count of late and early cobblestone area-forming cells in the peripheral blood increased. The number of endogenous committed precursor cells in the peripheral blood increased in all groups of patients with chronic myeloproliferative diseases and, particularly, in patients with idiopathic myelofibrosis and chronic myeloid leukemia. Functional characteristics of immediate descendants of hemopoietic stem cells probably reflect the level of damage and attest to the existence of various mechanisms underlying the defect of the hemopoietic stem cell during chronic myeloproliferative diseases. PMID- 18399276 TI - In vitro effects of thyroliberin on structural state of plasma membranes in mouse brain and liver. AB - The spin probe method was employed to study in vitro the effect of regulatory peptide thyroliberin on structural state of surface (0.8 nm) and deep (2 nm) lipid layers of the plasma membranes in mouse liver and brain. Thyroliberin in a concentration range of 10(-3)-10(-18) M enhanced structural order of surface lipids, the maximum effect was observed at 10(-9)-10(-10) M. The dose-effect dependencies for microviscosity of deep lipids were nonlinear and had 3 extrema at 10(-4)-10(-7) M, 10(-9) M, and 10(-14)-10(-16) M. The greatest changes in lipid microviscosity produced by 10(-9) M thyroliberin are explained by lipid receptor interaction. PMID- 18399277 TI - Effect of antenatal hypoxia on age-specific dynamics of ECG parameters and content of biogenic amines in the central nervous system. AB - We studied the effect of acute antenatal hypoxia during the stages of progestation and early organogenesis on some ECG parameters and level of biogenic amines in brain structures in rats. The effect of acute hypoxic exposure during the organogenesis period on the studied parameters was more pronounced than the effect of acute hypoxic exposure during the progestation period. The shift of the autonomic balance towards the sympathetic regulation of cardiac activity is linked with increased content of biogenic amines in the brain stem and cortical structures. PMID- 18399278 TI - Dynamics of LPO products and oxidative modification of proteins in human brain during postnatal development. AB - Opposite changes in the content of LPO products and products of oxidative modification of proteins were detected in human brain structures in the course of postnatal development. A clear-cut ontogenetic reduction of LPO products was observed in field 17 of the cortex, archicortex structures, and in the hypothalamus. Age-specific increase in the levels of products of oxidative modification of proteins was recorded in all compartments of the brain; it peaked by the age of 12-21 years and was most pronounced (4-6-fold) in the visual cortex, hippocampus, diencephalic and pontobulbar compartments of the brain. PMID- 18399279 TI - Study of the mechanism of hemostatic effect of desmopressin. AB - Intravenous injection of 1 microg/kg desmopressin to rabbits not only accelerated clotting of arterial blood (maximally by 59.16 +/- 8.53% after 1 h), but also increased the number of microvesicles containing the integral enzyme 5' nucleotidase in the arterial blood from the initial level of 36.26 +/- 8.08 ncat/liter to a maximum of 99.65 +/- 15.8 ncat/liter after 15 min. PMID- 18399280 TI - Oxidative status and citrate concentration in rat tissues during experimental hyperthyroidism and melatonin treatment. AB - Biochemiluminescence increased, while aconitate hydratase activity and citrate accumulation in tissues of the liver and heart and blood decreased in rats with experimental hyperthyroidism. These changes reflect activation of free radical oxidation, damage to enzyme molecules with reactive oxygen species, and impaired utilization of citrate under pathological conditions. Melatonin treatment during hyperthyroidism normalized aconitate hydratase activity and citrate concentration. Biochemiluminescence study showed that the effect of melatonin is related to antioxidant activity of this hormone, inhibition of free radical oxidation, and suppression of reactive oxygen species generation. PMID- 18399281 TI - Effect of N-terminal tripeptides of bombesin, litorin, and their analogue on body temperature and vasomotor responses. AB - Intranasal administration of bombesin caused hypothermia in rats maintained under cold conditions. N-terminal tripeptide of bombesin exhibits intrinsic vasomotor activity, while intranasal administration of its modified analogue produced a more potent hypothermic effect than intranasal bombesin. PMID- 18399282 TI - Self-administration of morphine by rats causes monoamine release in the anterior cingulate cortex. AB - Monoamine content in the microdialysate from the anterior cingulate cortex was measured in rats after injections and self-administration of morphine. Forced intraperitoneal injection of morphine did not lead to appreciable changes in the monoamine levels in the dialysate. Self-administration significantly increased monoamine levels in the extracellular space of the anterior cingulate cortex. Changes in catecholamine levels in the extracellular space of the anterior cingulate cortex correlated with the intensity of self-administration. The more morphine the animals injected to themselves, the greater was the increment in dopamine and norepinephrine levels. It seems that the increase of serotonin content in the anterior cingulate cortex did not depend on blood morphine concentration, but just reflected the fact of narcotic self-administration. The release of serotonin from nerve endings in the anterior cingulate cortex gradually increased at the beginning of the session, after which serotonin concentration started to decrease. The results indicate that monoamines are released from nerve endings in the anterior cingulate cortex only in response to self-administration of morphine. Catecholamines were released after each self administration of the narcotic, while serotonin release seemed to be associated with the general status of the animal realizing this behavior. PMID- 18399283 TI - Effect of magnesium chloride and magnesium L-aspartate on seizure threshold in rats under conditions of dietary magnesium deficiency. AB - We studied the effect of Mg-L-aspartate, MgCL2, and their combinations with vitamin B6, magneB6, and MgSO4 on seizure threshold in rats with dietary Mg2+ deficiency. Mg2+ deficiency was followed by a decrease in the threshold dose of corazol (from 79.20 to 49.48 mg/kg), shortening of the latency of the first jerk (by 33.6%, p=0.012), and reduction of the time to the first episode of clonic seizures (by 32.6%, p=0.011). Seizure threshold and latencies of the first jerk and first episode of clonic seizures increased over 3 weeks after peroral administration of Mg2+ salts. The combination of Mg2+ salts and pyridoxine was most effective in this respect. PMID- 18399284 TI - Comparative study of preventive and therapeutic effects of IEM-1966 and memantine in rats with experimental allergic encephalomyelitis. AB - We compared preventive and therapeutic effects of memantine, a selective blocker of NMDA-receptors, and IEM-1966, a blocker of both NMDA- and GluR1 AMPA receptors, on the model of acute experimental allergic encephalomyelitis. Memantine in high doses prevented the development of experimental allergic encephalomyelitis only in 10% rats, slightly (by 1.4-1.5 times) moderated the neurological disturbances, and shortened the duration of the disease. In far lower doses, IEM-1966 prevented the development of experimental allergic encephalomyelitis in 50% rats, while in the affected rats it decreased the severity of neurological disturbances and duration of the disease by 3-4 times. When applied during the clinical phase of the disease, IEM-1966 decreased the severity of neurological disturbances and duration of the disease by 2.0-2.5 times predominantly in rats with mild and moderate course of experimental allergic encephalomyelitis. PMID- 18399285 TI - Immnune response in mice of a new strain ASC (antidepressants sensitive catalepsy). AB - Inherited predisposition of ASC mice with depressive behavior to catalepsy was accompanied by a significant decrease in the immune response to sheep erythrocytes (compared to parent strains CBA and AKR). The degree of immunosuppression was highest on day 5 after immunization. PMID- 18399286 TI - Effect of chemotype on Escherichia coli interactions with bactericidal proteins. AB - We studied the effects of bactericidal proteins (lysozyme and lactoferrin) on endotoxin release from cell wall and inhibition of the growth of Escherichia coli colonies of different chemotypes. The structure of LPS core was found to be essential for the mechanisms of the interactions of the studied proteins with the cell wall. Cell viability after contact with cationic proteins is determined not by the amount of released LPS, but by the mechanism of damage to the cell wall. PMID- 18399287 TI - Direct evaluation of drug resistance parameters in gonococcus. AB - We carried out complex genetic analysis of clinical samples containing N. gonorrhoeae DNA, the genotype and profile of drug resistance of this agent were evaluated. Changes in genes responsible for the formation of N. gonorrhoeae resistance to penicillins, fluoroquinolones, and spectinomycin were detected during minisequencing with subsequent MALDI-TOF mass spectrometry. The sensitivity of gonococcus was evaluated directly in the clinical sample without culturing. PMID- 18399288 TI - Combined effect of cisplatin and lymphokine-activated killer cells on A549 cells of non-small cell lung cancer. AB - We studied the ability of lymphokine-activated killer cells to lyse A549 human non-small cell lung cancer cells after preincubation with cisplatin. Lymphokine activated killer cells obtained after incubation of human blood lymphocytes with interleukin-2 were characterized by high expression of natural killer cell antigens and activation molecules. Lymphokine-activated killer cells produced potent cytotoxic effect on intact A549 cells and lysed tumor cells survived after treatment with cisplatin in concentrations of IC50 and IC30. Cisplatin in noncytotoxic concentrations did not increase lytic activity of lymphokine activated killer cells. PMID- 18399289 TI - Age-related differences in expression of signal differentiation factors for human thymic epithelial cells. AB - Expression of transcription proteins PAX1, Hoxa3, and TLP regulating differentiation of thymic epithelial cells is detected in human thymus starting from gestation week 22 until the age of 95 years. Expression of transcription factors significantly decreased during aging. Apart from the decrease in the expression of signal differentiation factors in cultured thymic epithelial cells, proliferative activity of T lymphocytes cocultured with thymic epithelial cells also decreased in aging cultures, which demonstrated the important regulatory effect of transcription proteins on maturation and maintenance of T lymphocytes. Taking into account the important role of transcription proteins in the regulation of proliferation and function of T lymphocytes, whose number sharply decreases during aging, the maintenance of the level of expression of transcription factors during aging is a promising trend in modern biogerontology. PMID- 18399290 TI - Effect of laser radiation on production of reactive oxygen species in the blood of patients with chronic obstructive pulmonary disease. AB - The effect of laser radiation on generation of reactive oxygen species in the whole blood from patients with chronic obstructive pulmonary disease was studied by in vitro recording of luminol-dependent chemiluminescence. Laser irradiation of the blood from patients with increased production of reactive oxygen species decreased the microbicidal potential of cells. In patients with low generation of reactive oxygen species and normal potential of cells, laser exposure increased production of O2 metabolites. Laser radiation had little effect on chemiluminescence of the blood in patients with low generation of reactive oxygen species and decreased functional activity of cells. PMID- 18399291 TI - Autoantibodies to myosin light chains in the blood as early marker of myocardial injury after aortocoronary bypass surgery. AB - Serum concentration of autoantibodies to myosin light chains was measured after resumption of the bloodflow recovery in patients who underwent hypothermic aortocoronary bypass surgery. The patients were divided into 3 groups according to postoperative hemodynamic parameters and degree of myocardial injury. The studies showed significant differences in the concentrations of autoantibodies to myosin light chains between the groups. High correlation was shown between the duration of aorta clamping and concentration of autoantibodies to myosin light chains. Some factors characterizing initial severity of the disease can modulate blood concentration of autoantibodies to myosin light chains. PMID- 18399292 TI - Parameters of estrous cycles in albino rats normally and after injection of xenogenic cerebrospinal fluid. AB - We studied biological effects of xenogenic cerebrospinal fluid. Injection of the liquor to albino female rats during different periods of postnatal ontogeny induced changes in the dynamics of estrous cycles. Changes in the picture of vaginal smears, duration of estrous cycles, and proportion between their phases were found. PMID- 18399293 TI - Effects of different modes of interval hypoxic training on morphological characteristics and antioxidant status of heart and lung tissues. AB - The effects of various modes of interval hypoxic training differing by the intensity and duration of hypoxic exposure on the morphology and antioxidant status of the heart and lung tissues were studied. Interval hypoxic training mode with more severe, but shorter hypoxic component led to the prooxidant/antioxidant imbalance in the myocardial and lung tissues, which was paralleled by significant disorders in their morphology and function. Moderate hypoxic exposure of different duration promoted the maintenance of optimum antioxidant homeostasis and development of compensatory adaptive changes in tissue structure. PMID- 18399294 TI - Ultrastructural changes in the epithelium in asthma associated with gastroesophageal reflux. AB - Asthma concomitant with gastroesophageal reflux is regarded in the context of systemic multifactorial primary degenerative process, based on discoordination of neurohormonal regulation and systemic metabolic and degenerative changes. Degeneration and atrophy of the structural components of the bronchial tree and gastric wall are the leading phenomenons determining the organ dysfunction. PMID- 18399295 TI - Histomorphological characteristics of accelerated healing of acetate ulcers under the effect of glyprolines. AB - Glyprolines (PGP, GPG, GPGP, PGPGP, and GPGPGP) modulated histomorphological characteristics of acetate ulcers. They accelerated healing of acetate ulcers, promote complete differentiation of the surface epithelium and glands in the gastric mucosa, contributed to the appearance of a considerable number of fibroblasts at the site of the regenerating mucosa, and significantly decreased the count of macrophages. PMID- 18399296 TI - Coronarographic and pathomorphological characteristics of unstable atherosclerotic plaques in acute coronary syndromes. AB - The proximal segments of the main coronary vessels are the most often localization of "soft" unstable atherosclerotic plaques. The maximum number of plaques developed in the anterior descending branch of the left coronary artery. Pathognomonic relationship was found between the type of these plaques (with ulceration, rupture, thrombosis) and certain acute coronary syndrome. It was shown that the criteria of the plaque instability correlated with clinical manifestations of coronary syndromes. The bases and effects of "pathological" vascularization of unstable atherosclerotic plaques on the angioarchitectonics and hemodynamics of the heart were determined. Dissociation between myocardial vascularization degree and myocardial blood supply index was detected, which underlies the development of "unstable" myocardium in patients with acute coronary syndromes. PMID- 18399297 TI - On the accuracy of evaluations of temporal characteristics of visual perception. AB - We studied the accuracy of evaluations of the temporal characteristics of visual perception: critical frequency of light flickers, time of perception, and time of visual analyzer recovery. Results of comparison of the accuracy of evaluations are presented. PMID- 18399298 TI - Experimental validation for the use of a film on the basis of modified hyaluronic acid for prevention of postoperative peritoneal adhesions. AB - A film coating based on hyaluronic acid and 5-aminosalicylic acid was developed for the creation of anti-adhesion barrier. Application of the protective film in Wistar rats with modeled adhesion process in the abdominal cavity reduced the formation of peritoneal adhesions. PMID- 18399299 TI - Automating communication: using technology to increase effectiveness and profitability. PMID- 18399300 TI - Cadent iTero's digital system for dental impressions: the end of trays and putty? AB - Because conventional dental impression methods often are imprecise and subject to guesswork, and because these methods often are uncomfortable for the patient and expensive for the practitioner, the Cadent iTero system was created to help reduce or eliminate those negative aspects of dental impressions. The system's enhanced visualization and real-time analytical tools enable clinicians to adjust measurements before completing the intraoral digital scanning of dental patients. And because the iTero technology lends itself for use with all restorative procedures and materials via a network of dental laboratory partners, implantologists as well as other dentists interested in quality production of restorations can lower costs while assuring precise and patient-pleasing results. PMID- 18399301 TI - Dental implants and electrosurgery: Bident Bipolar Electrosurgical System. PMID- 18399302 TI - Social, demographic, clinical and lifestyle determinants of dental care visits in an urban sample of Portuguese adults. AB - PURPOSE: To describe the use of dental care services in an urban sample of adults from Porto, and to quantify the association between dentist visits and social, demographic, clinical and lifestyle factors. MATERIALS AND METHODS: Participants were selected by random digit dialling, and interviewed with a structured questionnaire to obtain information on socio-demographic, clinical and lifestyle variables. Crude, and age- and education-adjusted Odds Ratios (ORadj) were computed by unconditional logistic and multinomial logistic regression to quantify the association between the use of dental care and each exposure. RESULTS: In the year preceding the interview, 51.1% of the subjects visited a dentist at least once. Dental visits were less frequent in subjects aged > or =70 years compared to those aged 18-29 years (ORadj = 0.66, 95% Confidence Interval [CI]: 0.45-0.98), and increased with education (from 23.8% in subjects with 0-3 school years to 67.9% in those with > 12 years of education, p < 0.001 for trend). The use of dental care services at least once in the previous year was more frequent in white-collar workers (ORadj = 0.69, 95% CI: 0.52-0.91), when a private doctor was the usual source of medical care (ORadj = 1.38, 95% CI: 1.06 1.79) and in those who visited a medical doctor in the previous year (ORadj = 1.96, 95% CI: 1.57-2.45). Diabetics were less likely to seek dental care (ORadj = 0.67; 95% CI: 0.48-0.93). CONCLUSION: Nearly half the adults from Porto had not visited a dentist in the previous year. Education was the factor most strongly associated with dentist attendance. No differences were observed regarding the determinants of one or two or more dental visits in the previous year. PMID- 18399303 TI - Situational analysis of dental caries and existing oral health care of children and adolescents in Hubei Province, People's Republic of China: implications for change. AB - PURPOSE: (1) To evaluate trends in dental caries of children and adolescents of Hubei Province and Wuhan Municipality over the last two decades. (2) To conduct a situational analysis of existing oral health care and to recommend changes to improve this care for children. MATERIALS AND METHODS: An analysis was performed on data from national oral health surveys carried out in China at the provincial level, and data on 12-year-old children of Wuhan Municipality. A review of the literature was conducted on available and utilised oral health care facilities by children. RESULTS: In 1983, the mean decayed, missing and filled teeth (DMFT) of 12-year-olds in Hubei Province was 1.0, and in 1995 it was 0.6. The studies in Wuhan Municipality in 1993-1995 among 12-year-olds reported DMFT values ranging from 0.7-1.5, however different methods of caries assessment were performed in the various studies making comparisons difficult. The caries prevalence and the mean dmft score of 5-year-old children in 1995 in Hubei Province were 69% and 3.6, respectively. The percentage of untreated caries for all age groups in Hubei Province was very high. CONCLUSIONS: Analysis of epidemiological information over the last two decades does not provide any conclusive evidence supporting either an increasing or decreasing trend of dental caries in the child and adolescent population of Hubei Province and Wuhan Municipality. There is, however, a clear need to prevent the incidence of dental caries from increasing and to provide care to address the treatment needs of pre-school and school-age children. The promotion of twice-daily tooth brushing with fluoride toothpaste at maternal and child health care centres, and the establishment of daily tooth brushing with pea size amounts of fluoride toothpaste in the pre-school and primary school setting are recommended preventive strategies. Establishment of school-based oral health promotion should be accompanied with the provision of Atraumatic Restorative Treatment (ART) sealants and ART restorations which can be provided by dental nurses at dramatically lower costs than the traditional high-tech approach. PMID- 18399304 TI - Caries and periodontal disease: insights from two U.S. populations living a century apart. AB - PURPOSE: To compare caries experience and periodontal status between two U.S. populations that lived a century apart. MATERIALS AND METHODS: The University of Pittsburgh, School of Dental Medicine possesses a collection of skulls that was organised in the first half of the 20th century. Those individuals are likely to have lived in the northeast United States during the late 19th century and the early 20th century. The present study measured caries and periodontal disease morbidity in the collection of skulls and compared the data with current prevalence rates of these diseases in the United States (from the third National Health and Nutrition Examination Survey 1988-94). RESULTS: Periodontal health appears to have improved since this population resided, over 150 years ago. On the other hand, Americans between the ages of 16 and 39 appear to have the same prevalence of caries as a group of individuals that lived 150 years ago. CONCLUSIONS: The periodontal health of Americans has probably improved in the last 100 years. The individuals identified that were free of caries and periodontal disease among the group that lived during the 19th century may have some protective factor against these diseases. PMID- 18399305 TI - Development of a standardisation device for photographic assessment of dental fluorosis in field studies. AB - PURPOSE: This study evaluated the agreement between clinical and photographic assessment of dental fluorosis (DF) in 73 children during a field study, using portable equipment and a standardising device. MATERIALS AND METHODS: A dental exam was performed using the Tooth Surface Index of Fluorosis (TSIF). Photographs of the facial aspects of front teeth and from the occlusal aspect of the mandibular posterior teeth were taken. All photographs were taken following a standard operating procedure. Photographs of anterior and posterior teeth were coded and assigned a random study number. Examiners scored photographs as fluorosis cases or non-cases. RESULTS: Agreement among the clinical examiners ranged from good to excellent. Agreement was also good for the repeated photographic evaluations. In total, 22% of the children were diagnosed with DF during a clinical exam, and 19% were diagnosed with DF using the photographs. CONCLUSIONS: Development of this method aided in the photographic assessment of DF. The photographs obtained were a useful tool for documenting DF, to re-train examiners, and to determine intra- and inter-examiner agreement. PMID- 18399306 TI - Oral health status and oral hygiene habits of an adult Turkish population on dialysis. AB - PURPOSE: The present study was undertaken to evaluate the oral health status and oral hygiene habits of dialysis patients. MATERIALS AND METHODS: A total of 145 patients on dialysis were evaluated in two centres. Evaluation included oral examination of soft and hard tissues and completion of a questionnaire that was used to gather information about demographic, socioeconomic, medical and dental status and oral hygiene attitudes of patients. Awareness of the importance of oral health was also determined by four yes/no questions within this questionnaire. RESULTS: Of the 145 patients, 89 (61.4%) were presently candidates for renal transplantation. Nineteen patients (13.1%) were edentulous and nearly half of the patients (n=69, 47.6%) had 15 or fewer remaining teeth. Dentate patients who regularly brushed twice a day and flossed were few (n=18, 14.3%; n=3, 2.4%, respectively). In addition, plaque score was more than 50% in most of the dentate patients (93.7%). Gingival bleeding was reported by about two thirds of dentate patients (67.5%). None of the patients were referred to a dentist or were attending regular dental care. The percentage of patients who were aware of oral cavity-related infections and importance of oral hygiene following renal transplantation was very low (20.7% and 9.7%, respectively). CONCLUSIONS: Poor oral health, unsatisfactory daily oral hygiene habits and insufficient awareness of the importance of oral health is frequent among Turkish dialysis patients. Dentists and nephrologists should work in close cooperation in order to raise awareness and encourage regular dental controls. PMID- 18399307 TI - Smoking, tooth brushing and oral cleanliness among 15-year-olds in Tehran, Iran. AB - PURPOSE: To assess smoking, tooth brushing and oral cleanliness and their relationships among 15-year-olds in Tehran, Iran. MATERIALS AND METHODS: A cross sectional study based on World Health Organization criteria and the methods of the Second International Collaborative Study was carried out in autumn 2004 among 15-year-olds (n=502) in Tehran. Data were based on a self-administered questionnaire and a clinical dental examination. RESULTS: Smokers comprised 5% of the boys and 2% of the girls (p = 0.02). Smoking was more common among students of less-educated parents (50% vs. 30%, p < 0.05). Of all students, 26% reported twice-daily tooth brushing; those of higher socio-economic backgrounds and girls did so more frequently. Of the smokers, 11% reported no tooth brushing compared to 6% of the non-smokers. Oral cleanliness was good for 13%, moderate for 32%, and poor for 55%; the rates associated positively with female gender (p = 0.002), having higher-educated parents (p = 0.03), and reporting a higher frequency of tooth brushing (p < 0.001). Those students reporting twice-daily tooth brushing had less dental plaque and gingival bleeding (p < or = 0.01) on both anterior and posterior teeth. In multivariable analyses, the best predictors for a good level of oral cleanliness were female gender (OR = 2.0) or twice-daily tooth brushing (OR = 1.7). CONCLUSION: Oral cleanliness and tooth brushing among 15-year-olds were at poor levels, particularly among boys. Such poor levels call for intensive attempts to enhance rates of twice-daily tooth brushing and to improve its quality. For this age group, anti-smoking purposes should be combined into school based oral health promotion programmes as well. PMID- 18399308 TI - Risk-based approach in preventive practice among Iranian dentists. AB - PURPOSE: To study risk-based preventive practice among Iranian dentists. MATERIALS AND METHODS: A questionnaire survey was conducted at two annual dental meetings in 2004 and 2005 in Tehran. Using a five-point Likert scale, respondents indicated their level of agreement with taking preventive measures including oral hygiene, use of fluoride, diet and dental check-up for a high- and a low-risk (HR and LR, respectively) hypothetical patient case. Respondent's smoking and activity in smoking cessation were enquired about as well. Of 1033 responding dentists, 980 (64% men) were eligible for this study. Statistical evaluation was by the chi-square test and logistic regression. RESULTS: The top four of the eight measures were instructions on tooth brushing and flossing, advice related to fluoridated toothpaste, and regular dental check-ups, with the choice 'fully agree' being more prevalent for the HR (74%-58%) than for the LR case (59%-41%). For the HR case, 45% of the respondents fully agreed with applying chair-side tooth cleaning, 41% with advice on diet modification, and 38% with advice on home use of sodium fluoride mouthwash. Of all respondents, 76% were nonsmokers and 56% reported that they always recommend their smoking patients to quit. Female gender and activity in professional reading were associated with higher levels of agreement for applying preventive measures to the HR case. Non-smoking was the strongest explanatory factor (OR = 3.6, 95% CI = 2.6-5.1) of dentist's higher involvement in smoking cessation. CONCLUSION: Risk-based preventive dental care should be emphasised and applied in order to maximise efficient use of resources. PMID- 18399309 TI - The drop technique: a method to control the amount of fluoride dentifrice used by young children. AB - PURPOSE: The purpose of this study was to evaluate the amount of dentifrice applied to the toothbrush by school children using a liquid dentifrice (drop technique), when compared to toothpaste. MATERIALS AND METHODS: A total of 178 school children (4-8 years old) from two cities in Brazil (Bauru and Bariri) participated in the present two-part crossover study. Children from Bauru received training regarding tooth-brushing techniques and use of dentifrice before data collection. In each phase, the amount of toothpaste or liquid dentifrice applied by the children to the toothbrush was measured, using a portable analytical balance (+ 0.01 g). Data were tested by analysis of covariance (Ancova) and linear regression (p < 0.05). RESULTS: The mean (+/- standard deviation) amounts of toothpaste and liquid dentifrice applied to the toothbrushes for children from Bauru were 0.41 +/- 0.20 g and 0.15 +/- 0.06 g, respectively. For children from Bariri, the amounts applied were and 0.48 +/- 0.24 g and 0.14 +/- 0.05 g, respectively. The amount of toothpaste applied was significantly larger than the amount of liquid dentifrice for both cities. Children from Bariri applied a significantly larger amount of toothpaste, when compared to those from Bauru. However, for the liquid dentifrice, there was no statistically significant difference between the cities. A significant correlation between the amount of toothpaste applied and the age of the children was verified, but the same was not found for the liquid dentifrice. CONCLUSION: The use of the drop technique reduced and standardised the amount of dentifrice applied to the toothbrush, which could reduce the risk of dental fluorosis for young children. PMID- 18399310 TI - The relationship between smoking history, periodontal screening and recording (PSR) codes and overweight/obesity in a Mississippi dental school population. AB - PURPOSE: To examine the risks for being overweight (Ow) or obese (Ob) in subjects with elevated Periodontal Screening and Recording (PSRTM) Codes (> or =1) (with smoking as a modifier) in a population of dental school patients located within a region with a high incidence of Ow, Ob, diabetes mellitus (Db), and cardiovascular disease (CVD). MATERIALS AND METHODS: 1098 patient files were studied and data were grouped by average PSR Codes, smoking history, and body mass index (BMI). Data were compared by factorial ANOVA, the Mann-Whitney U test, and chi-square analysis using PSRTM Code as an exposure for BMI, Db, or CVD, with smoking as the effect modifier. Data were then analysed by adjusted multivariate logistic regression to determine the risks for being Ow or Ob. RESULTS: Subjects with an average PSRTM Code > or =1 and a smoking history did not have a significantly increased risk for either Db or CVD. Non-smoking subjects with an average PSR Code =3 had a significantly higher risk for being Ow; those with an average PSR Code of 2-4 had a significantly higher risk for being Ob (p < 0.05). In contrast, only smokers with an average PSR Code =4 had a significant risk for being either Ow or Ob. CONCLUSION: Our data suggest that smoking may not directly increase the risk factors for Db or CVD in subjects with an average PSRTM Code > 1 (confirming the 'Smoker's Paradox'), but could be an indirect risk factor for these diseases as a consequence of its effect on body weight. PMID- 18399311 TI - Higher elastase activity associated with lower IL-18 in GCF from juvenile systemic lupus patients. AB - PURPOSE: Our aim was to evaluate the expression of interleukin-18 (IL-18), interleukin-l-beta (IL-1beta) and the amount of elastase activity in gingival crevicular fluid (GCF) from inflamed gingival sites in patients with juvenile systemic lupus erythematosus (JSLE), and compare these to the expression in GCF from inflamed sites in generally healthy controls. In addition, the local inflammation in periodontal tissues was related to systemic inflammation by the assessment of IL-18 levels in plasma. MATERIALS AND METHODS: GCF from 16 patients with JSLE and 14 controls were collected using a washing device. Elastase activity was measured with a specific substrate, and IL-18 and 11-1 were measured by ELISA. RESULTS: The percentage of visible plaque index, gingival bleeding index and attachment level were similar in JSLE and controls, while the percentage of probing depth greater or equal to 3 mm was significantly higher in the controls. The total amount of IL-1beta and IL-18 in GCF were significantly decreased in JSLE, while the total amount and the percentage of free elastase activity were significantly higher in JSLE when compared with the controls. The plasma levels of I1-18 and the erythrocyte sedimentation rate were significantly higher in JSLE patients. CONCLUSION: We found more active elastase in GCF from inflamed sites in JSLE patients even in the presence of significantly lower levels of IL-18 and IL-13. The increased elastase activity suggests a hyperactivity of neutrophils in JSLE, possibly generated by a priming effect caused by the higher plasma levels of IL-18 found in these JSLE patients. PMID- 18399312 TI - Malaria vaccine: a current perspective. AB - The observation that inactivated Plasmodium sporozoites could protect against malaria is about a hundred years old. However, systematic demonstration of protection using irradiated sporozoites occurred in the nineteen-sixties, providing the impetus for the development of a malaria vaccine. In 1983, the circumsporozoite protein (CSP), a major sporozoite surface antigen, became the first Plasmodium gene to be cloned, and a CSP-based vaccine appeared imminent. Today, 25 years later, we are still without an effective malaria vaccine, despite considerable information regarding the genomics and proteomics of the malaria parasites. Although clinical immunity to malaria has been well-documented in adults living in malaria endemic areas, our understanding of the host-immune responses operating in such malaria immune persons remains poor, and limits the development of immune control of the disease. Currently, several antigen and adjuvant combinations have entered clinical trials, in which efficacy against experimental sporozoite challenge and/or exposure to natural infection is evaluated. This review collates information on the recent status of the field. Unresolved challenges facing the development of a malaria vaccine are also discussed. PMID- 18399313 TI - Studies on mosquitoes breeding in rock pools on inselbergs around Zaria, northern Nigeria. AB - BACKGROUND & OBJECTIVES: Rainwater often collects in depressions on rocks to form pools that are ideal breeding sites of mosquito vectors of diseases. Knowledge on the existence of disease vectors in these remote and relatively inaccessible locations could improve epidemiologic understanding and control capabilities. This study identifies mosquito species, their relative abundance and physicochemical characteristics of breeding microhabitats in rock pools on four inselbergs in northern Nigeria. METHODS: Soup ladle dipper was used to obtain representative samples of larval mosquitoes breeding in 141 rock pools on four inselbergs. Physicochemical parameters (depth, electrical conductivity, pH, surface area, temperature and total dissolved solids) of the pools were determined. Larvae were preserved in 70% alcohol and identified microscopically to species using taxonomic keys. Statistical correlation analysis and ANOVA were used to test the associations between physicochemical parameters and mosquito abundance, and for differences amongst inselbergs. RESULTS: Of 2991 larvae, five species of mosquito distributed in three genera (Anopheles, Aedes and Culex) including Ae. vittatus (92.88%), An. ardensis (0.13%), An. distinctus (1.67%), An. wilsoni (0.13%) and Cx. ingrami (5.18%) bred in the rock pools, Up to five species occurred per pool in various conspecific and heterogeneric combinations. Except for Ae. vittatus, the physicochemical parameters of the pools correlate significantly with species abundance. CONCLUSION: Ae. vittatus, a potential vector of yellow fever in Nigeria breeds profusely in rock pools on inselbergs around Zaria. For comprehensive vector implication and control, rock pools should be amongst the habitats of focus in yellow fever epidemiology. PMID- 18399314 TI - Population dynamics of onchocerca volvulus microfilariae in human host after six years of drug control. AB - BACKGROUND & OBJECTIVES: Mass administration of ivermectin drug was carried out annually between 1995 and 2001 in three villages that were endemic for onchocerciasis in the Lower Cross River Basin, Nigeria. The aim of this study was to evaluate the population dynamics (dispersion patterns, distribution, prevalence and intensity) of Onchocerca volvulus microfilariae in their human host after six years of ivermectin treatment. METHODS: A total of 1014 subjects from three rural areas in Etung Local Government Area of Cross River State, Nigeria were screened for skin microfilariae using standard parasitological method of diagnosis. RESULTS: Ivermectin drug intervention had significantly reduced the prevalence of skin microfilariae (PMF) from 69.3% pre-control to 39.3% and community microfilarial load (CMFL) from 7.11 to 2.31 microfilariae per skin snip. Males (45%) were significantly (p <0.05) more infected than females (34%). Both microfilarial prevalence and intensity increased with age. Pearson correlation test between intensity and age was not significant (r = 0.37; p >0.05). The correlation between age-dependent parasite prevalence and mean abundance was also not significant (r = 0.42; p >0.05). The degree of dispersion as measured by variance to mean ratio (VMR), coefficient of variation (CV) and exponent 'K' of the negative binomial model of distribution showed that the parasite aggregated, clumped and overdispersed in their human host. The relative index of potential infection of each age group showed that adults between the age of 21 and 50 yr accounted for 52.7% of microfilariae positive cases. INTERPRETATION & CONCLUSION: Aggregated and overdispersion of O. volvulus observed in this study showed that active transmission could still be going on, because the tendency of the vector, Simulium damnosum ingesting more microfilariae was high due to the aggregated and overdispersed nature of the parasite with its host. PMID- 18399315 TI - Entomological survey of phlebotomine sand flies (Diptera: Psychodidae) in a focus of visceral leishmaniasis in central Iran. AB - BACKGROUND & OBJECTIVES: Visceral leishmaniasis (VL) is a major vector-borne disease in Iran. A focus of VL is present in Shahreza county, Isfahan province, central Iran. The main objective of this study was to determine the probable vectors in this area. METHODS: Sand flies were collected biweekly using sticky paper traps, CDC light-traps, and aspirators from outdoors as well as indoors. All female sand flies were dissected and identified. Promastigotes were inoculated to hamsters and detected by Nested PCR. Approximately 7528 sand flies representing 12 species were collected from April 2003 to October 2004. Phlebotomus (Phlebotomus) papatasi Scopoli, Phlebotomus (Larroussious) major Annandale and Sergentomyia (Sergentomyia) sintoni Pringle were the predominant species. Two percent of P. major was found with natural promastigote infections. INTERPRETATION & CONCLUSION: This is the first report of natural promastigote infection in P. major in central Iran. The activity of P. major started from April and ended in October with a peak in September. The parasites were identified as Leishmania infantum using standard PCR. P. major is a possible vector of leishmaniasis and is susceptible to DDT in this area. PMID- 18399316 TI - Malaria related knowledge, practices and behaviour of people in Nepal. AB - BACKGROUND & OBJECTIVES: The information on malaria related knowledge, practices and behaviour of the people of Nepal living in malaria endemic areas are essential to develop behavioural change communication messages and for producing policy to prevent and control malaria in the country. The objective of this study was to assess knowledge, practices and behaviour of the people living in malaria endemic districts and relate with malaria control policy in Nepal. METHODS: The paper utilizes data from a cross-sectional study of 1330 households conducted during December 2004 to April 2005 in rural areas of Jhapa, Kailali and Kanchanpur districts. The method used includes structured questionnaire and focus group discussions. RESULTS: The results revealed that 40% respondents were illiterates. Eighty-six percent respondents have heard about malaria but only 50% responded fever with chills as the sign and symptom of malaria. Seventy-three percent responded that mosquito bite causes malaria transmission and 74% respondents considered that malaria is the fatal disease but very few have knowledge that the treatment of malaria in time can save life. More than 50% did not have information on availability of free treatment of malaria in Nepal. Still 16% were found consulting traditional healers for the treatment. The outside sleeping habit was found in almost one fourth of the population mainly in summer season indicating no knowledge about prevention of malaria. Although bednet use practice was higher, only 4% had knowledge on insecticide impregnated bednets and 23% of them practicing it. Issues regarding the proper and regular use of bednets, the quality of the bednet and use of free treatment provided by the government, since these districts are reporting high incidence of malaria. INTERPRETATION & CONCLUSION: Health education must be taken into account for communities in malaria endemic areas to produce desired outcomes in malaria control. PMID- 18399317 TI - Chloroquine prophylaxis associated with high prevalence of Plasmodium falciparum pfcrt K76T mutation in people with sickle-cell disease in Benin City, Nigeria. AB - BACKGROUND & OBJECTIVES: High mortality and morbidity in sickle-cell disease has been associated with malaria infection especially in countries where chloroquine is used. Chloroquine resistance has been associated with the emergence of Pfcrt mutant genes. This study aimed at comparing the prevalence rate of Pfcrt T76 mutation in Plasmodium falciparum isolates from infected individuals with sickle cell disease and sickle-cell trait. This study was carried out in Benin City between the months of April and June 2006. This period is marked with high transmission rate of malaria. METHODS: The genotype of the subjects was screened using haemoglobin electrophoresis system and the P. falciparum. Pfcrt genotyping was carried out using PCR-restriction fragment length polymorphism (RFLP). RESULTS: Four hundred and twenty-four subjects comprising of 207 haemoglobin AA, 136 haemoglobin AS and 81 haemoglobin SS typed individuals were enrolled for this study. No significant difference existed in the prevalence rate of malaria in the three groups (p > 0.05). However, the prevalence rate of Pfcrt K76T mutant gene was higher in the haemoglobin SS genotyped individuals than the haemoglobin AA and AS subjects (p < 0.05). INTERPRETATION & CONCLUSION: An uncontrolled use of chloroquine has been incriminated as the major cause of chloroquine resistance in Nigeria. Therefore, rapid intervention measures are needed as a matter of urgency to curb the up rise in the prevalence of the chloroquine resistant genes in our environment. PMID- 18399318 TI - Distribution and seasonality of vertically transmitted dengue viruses in Aedes mosquitoes in arid and semi-arid areas of Rajasthan, India. AB - BACKGROUND & OBJECTIVES: Transovarial transmission of dengue virus is a crucial etiological phenomenon responsible for persistence of virus during inter-epidemic period of the disease. Distribution and seasonality of this phenomenon in disease endemic areas may contribute to explain emergence of dengue and its subsequent prevention. The study on seasonal and area distribution of transovarial transmission of dengue virus in Aedes aegypti, Ae. albopictus and Ae. vittatus has been made in desert and non-desert districts of Rajasthan, India from 2006 to 2007. The observations revealed role of different Aedes species in transmission and retention of dengue virus. METHODS: The larvae of Ae. aegypti, Ae. albopictus and Ae. vittatus were collected during each of the study seasons from rural and urban areas of three districts-Jodhpur, Jaipur and Kota. The larvae were collected from domestic and peri-domestic containers and from tree holes of peri urban foci such as gardens and parks and were reared into adults in the laboratory at room temperature. The laboratory reared adults were subjected to Indirect fluorescence antibody test (IFAT). The laboratory-reared adult mosquitoes showing positive IFA were treated as the sample showing vertically transmitted dengue virus. RESULTS: Pooled data for all the four seasons revealed maximum (15.7%) mosquito infectivity in Ae. albopictus followed by Ae. aegypti (12.6%) in Jodhpur district. In Jaipur district, Ae. vittatus showed highest infection (20%) of vertically transmitted virus followed by Ae. albopictus (18.7%) and least in Ae. aegypti (13.3%). In Kota district, pooled data for all the four seasons showed maximum vertical infection of mosquitoes in Ae. albopictus (14.2%). INTERPRETATION & CONCLUSION: Transovarial transmission of dengue virus by available vector species in a dengue endemic setting could be the key etiological phenomenon responsible for re-emergence of the disease from inter epidemic to epidemic phase of disease onset. The observations in the present study suggest that during winter season which is not the active transmission season of dengue in Rajasthan, Ae. albopictus has shown maximum percentage of vertically transmitted virus. Our observation substantiates with the earlier studies that how Ae. albopictus is horbouring virus during inter-epidemic period of dengue. Another important lead emerging through present study is the high mosquito infectivity of Ae. aegypti during summer and rainy seasons especially from desert districts, Jodhpur and semi district. This observation suggests that in Rajasthan, owing to tendency of overstorage of domestic water by the inhabitants, mosquito and vertically transmitted virus get pronounced during summer season which could precedes the active transmission season of dengue during following rainy season. PMID- 18399319 TI - In vitro bioequivalence study of nine brands of artesunate tablets marketed in Nigeria. AB - BACKGROUND & OBJECTIVES: The availability of numerous brands of artesunate in our drug market today places clinicians and pharmacists in a difficult situation of choice of a suitable brand or the possibility of alternative use. The aim of the present study was to predict the bioequivalence of nine brands of artesunate tablets marketed in Nigeria using in vitro tests. METHODS: The in vitro dissolution study was carried out on the nine brands of artesunate tablets using the basket method according to US Pharmacopoeia (USP) guidelines. Other general quality assessment tests like hardness and disintegration time were also determined. RESULTS: All the brands tested passed the British Pharmacopoeia (BP) standard for disintegration time. Only AT2, AT4, AT6 and AT9 passed the standard for hardness. There were significant differences in the dissolution profiles of the nine brands. All the brands except AT1, however, released >70% of artesunate within 30 min. Four of the brands AT5, AT6, AT7 and AT8 exhibited >90% dissolution in <10 min. The other brands AT1, AT2, AT3, AT4 and AT9 (innovator brand) have calculated similarity factors of 23.8, 59.8, 50, 54.8 and 100. INTERPRETATION & CONCLUSION: Based on the in vitro tests, AT5, AT6, AT7 and AT8 are considered bioequivalent and interchangeable, while AT2, AT3 and AT4 are considered bioequivalent and interchangeable with the innovator brand (AT9). AT1 has very low dissolution rate, which will likely result in poor bioavailability. The results show the need for constant monitoring of new brands of artesunate introduced into the drug market to ascertain bioequivalence and conformity with pharmacopoeia standards. PMID- 18399320 TI - Larvicidal activity of leaf extract of Millingtonia hortensis (Family: Bignoniaceae) against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti. PMID- 18399321 TI - Sunlight exposure enhances larval mortality rate in Culex quinquefasciatus Say. PMID- 18399322 TI - Knowledge on lymphatic filariasis and mass drug administration (MDA) programme in filaria endemic districts of Andhra Pradesh, India. PMID- 18399323 TI - Acalculous cholecystitis by P. falciparum in a 3-year-old child. PMID- 18399324 TI - Impact of maternal Plasmodium falciparum malaria and haematological parameters on pregnancy and its outcome in southeastern Nigeria. PMID- 18399325 TI - Gene Regulatory Network modelling: a state-space approach. AB - This study proposes a state-space model with control portion for inferring Gene Regulatory Networks (GRNs). The proposed model views genes as the observation variables, whose expression values depend on the current internal state variables and control variables, and views the means of clusters of gene expression as the control variables of the internal state equation. Bayesian Information Criterion (BIC) and Probabilistic Principal Component Analysis (PPCA) are used to estimate the internal states from observation data. The proposed approach is applied to two gene expression datasets. Computational results show that inferred GRNs possesses the characteristics of the real-life GRNs. PMID- 18399326 TI - Segmentation of short human exons based on spectral features of double curves. AB - This paper presents a new segmentation method based on spectral analysis to locate borders between short protein coding regions and non-coding regions. We formulate the innovative double curve representation of a DNA sequence and apply local three-codon measurement on the discrete Fourier spectral features at 1/3 frequency to identify short protein coding regions. The proposed spectral segmentation method based on double curves requires no prior knowledge of the DNA data. Our simulation results show that the proposed spectral method greatly improves the accuracy of identifying short coding regions in DNA sequences compared with the results obtained from the other methods that analyse DNA sequences directly. PMID- 18399327 TI - Temporal representation for gene networks: towards a qualitative temporal data mining. AB - Processing literature (i.e., text corpora) to capture gene regulation events is not easy and can be driven by the final data representation. We propose to build, manually, an example of temporal representation (whole gene networks for coat formation in Bacillus Subtilis). Our temporal representation is based on a generalised formal language theory (S-languages). We propose an algorithm to link bags of relations with representation, by ordering interactions. In this paper, starting from the network made manually from text data, we show that S-languages are quite relevant to encapsulate gene properties, and infer knowledge across timestamped gene relations found in texts. PMID- 18399328 TI - An integrative approach for biological data mining and visualisation. AB - The emergence of systems biology necessitates development of platforms to organise and interpret plentitude of biological data. We present a system to integrate data across multiple bioinformatics databases and enable mining across various conceptual levels of biological information. The results are represented as complex networks. Context dependent mining of these networks is achieved by use of distances. Our approach is demonstrated with three applications: full metabolic network retrieval with network topology study, exploration of properties and relationships of a set of selected proteins, and combined visualisation and exploration of gene expression data with related pathways and ontologies. PMID- 18399329 TI - A rule-based approach for RNA pseudoknot prediction. AB - RNA plays a critical role in mediating every step of cellular information transfer from genes to functional proteins. Pseudoknots are functionally important and widely occurring structural motifs found in all types of RNA. Therefore predicting their structures is an important problem. In this paper, we present a new RNA pseudoknot structure prediction method based on term rewriting. The method is implemented using the Mfold RNA/DNA folding package and the term rewriting language Maude. In our method, RNA structures are treated as terms and rules are discovered for predicting pseudoknots. Our method was tested on 211 pseudoknots in PseudoBase and achieves an average accuracy of 74.085% compared to the experimentally determined structure. In fact, most pseudoknots discovered by our method achieve an accuracy of above 90%. These results indicate that term rewriting has a broad potential in RNA applications ranging from prediction of pseudoknots to discovery of higher level RNA structures involving complex RNA tertiary interactions. PMID- 18399330 TI - Pain management in adult acute sickle cell pain crisis: a viewpoint. AB - BACKGROUND: The acute pain crisis of sickle cell disease is inadequately treated in many countries. OBJECTIVE: To present a simple protocol that controls acute pain in most adult patients within 72 hours, based on our experience in an area where sickle cell disease is highly prevalent. METHODS: Patients aged 14 years and above with sickle cell disease presenting with pain crisis are treated initially in the Emergency Room. Those responding adequately are discharged home on oral analgesics while those with persisting pain after 6 hours are treated further, for up to 72 hours, in an Observation Unit attached to the Emergency Room. Narcotic analgesics are administered regularly for the first 24 hours. RESULTS: Using this protocol, acute pain crisis could be terminated or controlled in over 80% of patients within 72 hours. CONCLUSION: Regular intravenous narcotic analgesia for the initial 24 hours, supplemented by oral analgesia, is useful for adult patients with severe acute sickle cell pain crisis when the pain is inadequately controlled by initial Emergency Room treatment. There may be gender differences in pain perception and response in acute sickle cell pain crisis. PMID- 18399331 TI - Relationship between waist circumference and cholesterol in Central Africans with congestive heart failure. AB - BACKGROUND: There are controversies as to what the traditional risk factors for coronary heart disease should be in sub-Saharan Africa. OBJECTIVE: To assess the relationship between cholesterol and cardiovascular disease risk factors including Helicobacter pylori infection in black Africans with congestive heart failure. METHOD: A cross-sectional and observational study of 48 men and 52 women. RESULTS: Congestive heart failure was associated with abdomal obesity, hypertension, chronic renal failure, moderate levels of low HDL-C, excessive alcohol intake and hyperuricaemia, but low levels of cholesterol and triglycerides. TC was related by Univariate analysis with red cells, glucose, weight, waist circumference with HDL-C, CRP, fibrinogen and IgG antibodies against H pylori. Multivariate analysis revealed that waist circumference (B=0.688) and HDL-C (B=0.826) were the significant determinants of TC. There was a respective U-shaped relationship between CVD (P>0.05), chronic renal failure (P<0.05) H. pylori-induced chronic gastritis (P<0.05) and the HDL-categories. Ischemic stroke and myocardial infarction were significantly (p<0.05) associated with low HDL-C, respectively. Clinical insulin resistance (P<0.01) was predominantly more commonin the intermediate HDL-C category than in low and high HDL-categories. There was an inverse relation between lower TC: HDL-C ratio, high HDL-C and abdominal obesity/ insulin resistance in men. H. pylori gastritis was positively related to higher TC: HDL-C ratio in both men and women. CONCLUSION: Preventive measures, more studies on the interplay between HDL-C level and its function and a specific ethnic dfinition of metabolic syndrome in the African are needed. PMID- 18399332 TI - Molecular characterisation of Pseudomonas aeruginosa recovered in the Buea health district of Cameroon: implications for nosocomial spread. AB - BACKGROUND: Pseudomonas aeruginosa is a ubiquitous gram-negative pathogen with a propensity to cause opportunistic infections in humans. Different strains of the organism could colonise patients heralding a wide spectrum of P. aeruginosa infections in the environment. OBJECTIVE: To analyse isolates of P. aeruginosa from clinical and environmental samples using the Polymerase Chain Reaction (PCR) to establish strain relatedness. METHODS: Fifty-two strains of the organism were isolated from wound swabs, urine, sputum of patients and environmental samples from the hospital environment using standard microbiological techniques and ethical consideration. Genomic DNA of the isolates was amplified with primers AF1 (5'-AGA GTT TGA TCC TGG CTCA-3') and 1541R (5'-AAG GAG GTG ATC CAG CC-3'). RESULTS: At least two bands were observed in all isolates typed and band sizes ranged from 0.07 - 1.5kb. The strains were genetically diverse, displaying profiles of 2 - 6 bands between 0.07 - 1.5kb. CONCLUSION: The study demonstrates that strain diversity could be discerned between strains of P. aeruginosa, circulating in the environment of Buea, a finding which has important epidemiological and clinical significance bearing in mind that this pathogen is highly incriminated in nosocomial infections with attendant social implications. This therefore calls for more attention in the diagnosis and management of P. aeruginosa infections in the environment of Buea, Cameroon. PMID- 18399333 TI - Infection control audit and potential sources of infection of a Nigerian eye unit. AB - BACKGROUND: The prevalence of post-surgical eye infections in Nigeria has not been documented. However, anecdotal reports suggest that post-operative endophthalmitis is fairly common in Lagos. OBJECTIVES: This study was done to investigate the sources of post-operative eye infections and the preparedness of the hospital to prevent such infections. METHODS: A bacteriological survey of all eye surgical procedures between September 2004 and June 2005 was performed. Relevant samples were cultured pre- intra and post operatively as required. In the eye clinic, and operation theatre, infection control procedures and practices were audited using a pre-designed questionnaire. RESULTS: Out of 207 procedures performed, there was no case of post -operative eye infection but potential sources of infections identified included nares of staff, surgical packs, theatre bed, hands of surgeons and conjunctivae of patients. Running water and soap were available for hand washing. There was no specific provision for drying hands in the clinic and staff were observed to carry out hand wash considered ineffective. Staff showed good knowledge of disinfectants and antiseptic use. In the theatre, the staff who handle the steam sterilizer were not formally trained on its use. Although sterile packs were appropriately stored they were not dated. Appropriate types of waste bins were not in use and wastes were not segregated appropriately. Ophthalmic equipment were not adequately disinfected in between patients and use of eye drops for dilatation was communal. CONCLUSION: Potential sources of post operative eye infections were identified. Areas of deficiencies in infection control practices, which require proper infection control policies, include handwashing facilities and practices, sterilization procedures, disinfection of ophthalmic equipment, linen management and waste handling. PMID- 18399334 TI - Dental caries trend among adolescents in Lagos, South-West Nigeria. AB - BACKGROUND: The past few decades have witnessed a changing trend in dental caries prevalence among Nigerians; thus the need to investigate the current trend of caries among adolescents Nigerians. OBJECTIVE: To determine the prevalence and severity of dental caries among adolescents in Lagos, South-west of Nigeria. METHODS: The World Health Organization (WHO) methodology for basic oral health surveys was employed. The study population was 11 to 16 year-old adolescents from primary and secondary schools in Surulere, Lagos, Nigeria. Surulere local government area (LGA) of Lagos State, South-West Nigeria was selected as the location of this study as the LGA is quite cosmopolitan having Nigerians of all walks of life. Information on sex, age, socioeconomic status and ethnicity was obtained. The dentition of subjects was examined for dental caries, fillings and missing teeth. RESULTS: Six hundred adolescents of age 11 to 16 years were studied. A high caries-free prevalence of 457 (76.2%) was found among the subjects. Mean(SD) DMFT (decayed, missing, filled teeth) of 0.72 (1.67) and mean DMF% (decayed, missing, filled per cent) of 2.62 (6.17) were recorded among the study population. Older adolescents had higher caries erates than the younger ones. More females than males had caries while the socioeconomic status did not seem to influence caries experience. DT (decayed teeth) was the major contributor of the DMFT index. Among the ethnic groups the Igbo and Edo/Delta had the highest caries experience while the Efik/Ibibio, Yoruba and Hausa had the lowest. A restorative index of 1% was recorded, showing poor utilization of dental health care services by the population. CONCLUSION: This study shows an increasing caries trend when compared to previous Nigerian studies despite the mean DMFT of 0.72 being better than the WHO target of 1.5DMFT. Ready availability of sweets and confectionaries to all classes of people in the country compounded by poor knowledge of and utilization of preventive and restorative oral health care services may be contributory to the current trend. PMID- 18399335 TI - Perception of cigarette smoking and advertisement among senior secondary school students in Ibadan, Southwestern Nigeria. AB - BACKGROUND: Cigarette smoking among youths is increasingly a public health concern. OBJECTIVE: This study sought to determine the prevalence of smoking among secondary school students and to assess their perception of smoking and cigarette advertisement. METHODS: A cross sectional study was conducted among 1,223 senior secondary school students in Ibadan, Southwest Nigeria. RESULTS: The prevalence of smoking among this population was 41 (3.4%) with 115 (9.4%) reporting that they had ever smoked. Mean age of smoking initiation was 14.2 years. Smoking was greater among males than females and in higher secondary school classes. Only 700 (57%) of students knew that smoking was harmful to their health and 477 (39%) reported that they had been taught about the harmful effects of smoking in the school teaching programme. Twenty-six percent of students had an item with a cigarette brand name on it and 19% had been offered a free cigarette by a cigarette representative. One hundred and twenty-two (10%) of the students indicated that they will smoke when they leave school. CONCLUSION: Although the prevalence of smoking among students was low, exposure to cigarette advertisements may encourage them to smoke. PMID- 18399336 TI - Evaluation of Alvarado score as an admission criterion in patients with suspected diagnosis of acute appendicitis. AB - BACKGROUND: Patients with right iliac fossa pain present a diagnostic challenge to emergency room doctors when equivocal signs are present. OBJECTIVE: To evaluate Alvarado score as an admission criterion in patients with suspected diagnosis of acute appendicitis. METHODS: This was a prospective study of 100 consecutive patients who presented to the an emergency department with right iliac fossa pain and suspected diagnosis of acute appendicitis between January 2004 and December 2004. They were assessed usingAlvarado score. Main outcome measures in relation to these scores were, patient's admission, patient's discharge, and whether patient had appendicitis or not. RESULTS: The range of scores was 1-10. Seventy-four patients were admitted while 26 were discharged. Thirty-four patients with confirmed diagnosis of acute appendicitis had scores ranging from 5-10. No patient with score less than 5 had appendicitis. The sensitivity and negative predictive value reached 100% at scores below five, while specificity and positive predictive value reached 100% at the score of 10. CONCLUSION: The Alvarado score can be used as an objective criterion in selecting for admission patients with suspected diagnosis of acute appendicitis. Patients with scores of 4 and below do not require admission since they turn out not to have acute appendicitis. PMID- 18399337 TI - Efficacy and safety of misoprostol in induction of labour in a Nigerian tertiary hospital. AB - BACKGROUND: Misoprostol - a stable prostaglandin E1 analogue- is effective and safe in the induction of labour. There is paucity of information about the use of misoprostol for labour induction in Nigeria. OBJECTIVE: To evaluate the efficacy of misoprostol in the induction of labour in the third trimester. METHODS. Consecutive patients for induction of labour were randomized into misoprostol or oxytocin study groups. The misoprostol group received intravaginal 50 microg 6- hourly to a maximum of four doses. Those in the oxytocin group received a maximum of 48 iu/min. Outcome measures included induction-delivery interval, mode of delivery, Apgar score, perinatal death and maternal complications. RESULTS: Sixty two patients were recruited into the study-34 received misoprostol while 28 received oxytocin. The modal gestational age and Bishop score prior at induction were >36 weeks and 5-7 respectively. Hypertension in pregnancy was the commonest indication for induction of labour followed by prolonged pregnancy. The overall induction-delivery interval was 12.2 +/- 5.2 hours; Misoprostol v oxytocin, mean(range): 12.1(7-27) vs 12.3(4-27) hours, p = 0.88). There were no significant differences in the mean Apgar score and perinatal mortality rate in the two study groups. There were two cases of primary postpartum haemorrhage in the oxytocin group but none in the misoprostol group. One case of ruptured uterus was encountered in the misoprostol group. No case of maternal mortality was recorded. Four patients in the misoprostol group had minor side effects mainly nausea and vomiting. CONCLUSION: The efficacy of misoprostol in the induction of third trimester labour is comparable to oxytocin. The risk of ruptured uterus associated with misoprostol appears higher than that of oxytocin in the induction of labour. Further studies are needed to verify this observation in our setting. PMID- 18399338 TI - The effect of quinine and ascorbic acid on rat testes. AB - BACKGROUND: We have previously demonstrated that quinine is a testicular toxicant in Sprague-Dawley rat. OBJECTIVE: To describe the changes in the testicular levels of testosterone and lipid peroxidation secondary to quinine and ascorbic acid administration in rats. METHODS: Twenty male Sprague-Dawley rats per group were assigned to one of three treatment groups: 0 mg quinine and 0 mg ascorbic acid/kg body weight (control); 10 mg quinine/ kg BW; and 10 mg quinine plus 0.1 mg ascorbic acid/kg BW. Rats were intramuscularly administered their respective doses of quinine five days in a week and ascorbic acid three days in a week for eight weeks. All the animals were sacrificed at the end by decapitation. Seminal analysis was performed on tubular fluid from caudal epididymides. Evaluations were made for testicular levels of testosterone and lipid peroxidation through malondialdehyde (MDA). Testicular specimens were also processed for histology under light microscopy. RESULTS: Quinine significantly (p < 0.01) increased free radicals (from elevation of MDA) and decreased testosterone in the testis compared with those of the control group and those treated with a combination of quinine and ascorbic acid. The semen of rats treated with only quinine demonstrated a significantly (p < 0.001) lower sperm concentration and motility compared to the controls and those treated with quinine plus ascorbic acid. Microscopic examination of cross-sections of seminiferous tubules also showed that ascorbic acid partially protected against quinine -induced testicular effects. CONCLUSION: Ascorbic acid has beneficial effect and protects against quinine-induced testicular reduction of testosterone. PMID- 18399339 TI - Childhood Wilms' tumour: prognostic factors in North Central Nigeria. AB - BACKGROUND: Childhood Wilms' tumour or nephroblastoma represents one of the challenges for paediatric oncologists in developing countries. OBJECTIVE: To review the clinical characteristics and outcome of management of childhood nephroblastoma in North- Central Nigeria. METHODS: The clinical, operative and histological records of children aged 15 years and below, that were managed for Wilms' tumour at the Jos University Teaching Hospital between 1998 and 2005 were retrospectively reviewed. RESULTS: There were 32 children (M:F=1.9:1) with histologically confirmed nephroblastoma seen over the 7-year period. Their median(range) age was 4 (3-15) years. The patients invariably presented with a palpable abdominal mass, but haematuria was exceptional. The neoplasm tended to be larger on average than those reported previously among Caucasian children. At presentation, 1 (3.1%) patient was in stage I, 8 (25%) stage II, 11 (34.4%) stage III and 12 (37.5%) stage IV. About 72% of the patients presented with stage III IV disease. Poorly differentiated neoplasm was more common in male than in female patients. Nephrectomy and chemotherapy were the modality of treatment. Fifteen (46.9%) of the patients received little or no induction chemotherapy due to unavailability of drugs while only 12 (37.5%) received the prescribed maintenance treatment with the remainder getting erratic or no treatment. Overall, only 43.8% were alive between 1 and 9 months (median: 6 months) of follow-up period, but there was no survivor at two years after treatment. CONCLUSION: Childhood nephroblastoma has a high mortality rate in north central Nigeria because of late clinical presentation with advanced disease, poor availability of cytotoxic drugs and frequent interruptions in treatment and inadequate follow-up. PMID- 18399340 TI - Urinary bladder cancer and schistosomiasis in North-Western Nigeria. AB - BACKGROUND: An unusually high prevalence of bladder cancer was noticed by clinicians and in the cancer registry of Usmanu Danfodiyo University Teaching Hospital, Sokoto. Several areas of this region were also known to be endemic for urinary schistosomiasis. OBJECTIVE: To determine the epidemiological characteristics of bladder cancer in the region and to assess the impact of schistosomiasis on these cases. METHOD: Retrospective review of clinical and histopathogical records of bladder cancer cases seen at Usmanu Danfodiyo University Teaching Hospital, Sokoto from January 1999 to December 2004. RESULT: One hundred and thirty three cases satisfied our criteria for inclusion. There was a 4.7 fold rise in the number of bladder cancer cases between 1999 and 2004. The male to female ratio was 11.1:1.0. The mean age was 46.0 years and ranged from 20 to 82 years. Majority, 107 (80.5%) were farmers and fishermen from regions of the distribution of surrounding river or their smaller tributaries. Squamous Cell Carcinoma comprised 65.1% of histologically verified cases and in 50% of Squamous Cell Carcinoma; there was histological evidence of chronic urinary schistosomiasis. CONCLUSION: Bladder cancer is a common malignancy in Sokoto, North-Western Nigeria. The association with chronic urinary schistosomiasis is very strong and the hospital incidence appears to be rising. PMID- 18399341 TI - Clinical comparison of cardiorespiratory effects during unilateral and conventional spinal anaesthesia. AB - BACKGROUND: Spinal anaesthesia is widely employed in clinical practice but has the main drawback of post-spinal block hypotension. Efforts must therefore continue to be made to obviate this setback OBJECTIVE: To evaluate the cardiovascular and respiratory changes during unilateral and conventional spinal anaesthesia. METHODS: With ethical approval, we studied 74 American Society of Anesthesiologists (ASA), physical status class 1 and 2 patients scheduled for elective unilateral lower limb surgery. Patients were randomly allocated into one of two groups: lateral and conventional spinal anaesthesia groups. In the lateral position with operative side down, patients recived 10 mg (2mls) of 0.5% hyperbaric bupivacaine through a 25-gauge spinal needle. Patients in the unilateral group were maintained in the lateral position for 15 minutes following spinal injection while those in the conventional group were turned supine immediately after injection. Blood pressure, heart rate, respiratory rate and oxygen saturation were monitored over 1 hour. RESULTS: Three patients (8.1%) in the unilateral group and 5 (13.5%) in the conventional group developed hypotension, P= 0.71. Four (10.8%) patients in the conventional group and 1 (2.7%) in the unilateral group, P= 0.17 required epinephrine infusion to treat hypotension. Patients in the conventional group had statistically significant greater fall in the systolic blood pressures at 15, 30 and 45 minutes when compared to the baseline (P= 0.003, 0.001 and 0.004). The mean respiratory rate and oxygen saturations in the two groups were similar. CONCLUSION: Compared to conventional spinal anaesthesia, unilateral spinal anaesthesia was associated with fewer cardiovascular perturbations. Also, the type of spinal block instituted affected neither the respiratory rate nor the arterial oxygen saturation. PMID- 18399342 TI - [Psoas abscess in Cote d'Ivoire: a report of eighteen cases]. AB - BACKGROUND: Psoas abscess is a rarely encountered entity with a non specific clinical presentation resulting in delayed diagnosis and treatment. No published data exist on this disease in our country. OBJECTIVE: To describe the diagnostic and therapeutic features of psoas abscesses in Cote d'Ivoire. Methodes: A retrospective study of 18 psoas abscesses seen over seven years in two teaching hospitals. RESULTS: There were 10 women and eight men with a mean age of 35.7 years (range:16-62 years). The abscess was primary in 15 cases, secondary in three, right sided in 15 and left sided in three cases. Fever, abdominal pain, difficulties in walk, abdominal mass and psoitis were the main clinical signs. Ultrasonography allowed the diagnosis of psoas abscess in 14 cases and in the remaining 4 cases the diagnosis was done peroperatively. The germs were identified in 12 patients and were: Escherichia coli in 3 cases, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pneumoniae, Klebsiella pneumoniae in two cases each and Mycobacterium tuberculosis in one case. Patients were given antibiotics together with surgical drainage of the abscess (n=16) or percutaneous needle aspiration (n=2). Postoperative complications included parietal suppurations (n=5) and intraperitoneal abscesses (n=2). No death occurred. CONCLUSION: For psoas abscess in our practice, ultrasonography is a useful diagnostic tool and surgical drainage remains an effective therapeutic method. PMID- 18399343 TI - [Relevance of the systematic culture of the intraoperative swab and drain tip of Redon in orthopaedic-traumatology surgery]. AB - BACKGROUND: Post operative infections are more severe complications in bone surgery. The first stage culture on drain tip or intraoperative swab are not well known according to clear, open and aseptic orthopaedic surgery to predict wounds infections. OBJECTIVE: To show the place of the systemic bacteriological culture of an intraoperative swab and the proximal tip of the Redon in bone surgery. METHODS: This was a prospective continuous series of 92 interventions performed in the service of Orthopaedics Traumatology of Treichville University Hospital (Abidjan, Cote d'Ivoire). The lesions included were allocated into three groups based on the National Research Council classification. Group 1 consisted of 50 subjects with clean lesions and hyper clean. Group 2 was made up of 25 subjects with clean lesions contaminated or contamined ab initio while Group 3 consisted of 17 patients with the septic lesions. Fifty six men and 36 women with an average age of 36.9 years had two types of swabs culture. In the first type sample of intra operative haematoma or the pus before using antiseptic products was used; the second type of culture used the proximal tip of Redon at the time of its ablation. These two swabs were put in a sterile vial and sent to the same laboratory for culture. RESULTS: The overall sepsis rate was of 24(26,1%). The microbial population was dominated by the gram negative bacilli, bacilli positive intraoperative cultures were most frequent in the group 3. 15 (88,2%). The positivity of the culture of the Redon was high in the group 2 (32%) and in the group 3 (52.9%). There was a significant difference between these two groups of surgery. The sensitivity, the specificity, and the predictive values were low. For all groups, the reports of likelihood observed didn't permit to establish a relation of cause or effect between a positive culture and the occurrence of post operative infection. CONCLUSION: The gram negative bacilli were mostly observed on the culture of the site of infection. Although there was no significant relationshionship, it appears from the frequency that there may be a clinical link between the positive culture and open fracture. PMID- 18399344 TI - Staphylococcus lugdunensis endometritis: a case report. AB - BACKGROUND: Staphylococcus lugdunensis has been reported to cause several localized and blood stream infections, but not endometritis. OBJECTIVE: To desribe a case of Staphylococcus lugdunensis endometritis associated with premature rupture of membranes. CASE REPORT.A 39-year old woman presented with premature rupture of membrane (PROM) and underwent an emergency caesarean section at 40 weeks of gestation. Her endometritis was characterized by a foul odour and was so extensive that the baby was adherent to the endometrium and had to be separated by a gentle pull. In spite of these, neither the mother nor her baby suffered any adverse effect. The organism exhibited several unusual characteristics that are atypical ofstaphylococci. The baby did not develop any sepsis. The mother responded well to antibiotics and both were discharged home on the 4th postoperative day. CONCLUSION: To the best of our knowledge, this is the first report of this organism causing endometritis. PMID- 18399345 TI - Urethral fibrosis with pyonephrosis in a patient with situs inversus totalis: a case report. AB - BACKGROUND: Cases of a case of situs inversus totalis are very rare and therefore when encountered, they are likely to be missed or poorly managed. OBJECTIVE: To present a case of situs inversus totalis with a view to creating more awareness about this rare clinical entity. PATIENT AND METHOD: The patient, a 22 year old boy was referred from a private clinic after a failed and complicated attempted appendicectomy carried out by a general medical practitioner. The patient was admitted, resuscitated and had investigations done. Some of the investigations done were abdominal ultrasound scan, computerized axial tomography scan, a chest x ray, complete blood count, urinalysis and electrolyte and urea. RESULTS: Investigations showed features of situs inversus totalis. There was also bilateral hydronephrosis, worse on the right side. The patient also had features of renal failure. He died after three days on admission. Autopsy finding confirmed bilateral pyonephrosis which was worse on the right side. There was associated urethral fibrosis. Cause of death was urethral fibrosis resulting in obstructive uropathy, pyonephrosis and renal failure. CONCLUSION: Situs inverses totalis is rare but can psented in diverse ways and thus the diagnosis may be missed and the patient wrongly managed. A high index of clinical suspicion is required if we the diagnosis is to be made early and wrong treatments avoided. PMID- 18399346 TI - Macromastia and bilateralaxillary breast hypertrophy: a case report. AB - BACKGROUND: Breast hypertrophy presents at puberty or thereafter. It is a condition of abnormal enlargement of the breast tissue in excess of the normal proportion. Gland hypertrophy, excessive fatty tissue or a combination of both may cause this condition. Macromastia can be unilateral or bilateral. OBJECTIVE: To present a case of massive bilateral gigantomastia with huge bilateral hypertrophy of the axillary breasts. METHODS: Review of the prentation, clinical and investigative findings aswell as the outcome of surgical intervention of a young Nigerian woman with bilateral severe breast hypertrophy and severe hypertrophy of axillary breasts. RESULT: The patient was a 26-year-old woman who presented with massive swelling of her breasts and bilateral axillary swellings, both of six years duration.. In addition to the breast pathology, she also suffered significant psychological problems. The breast ultrasonography confirmed only diffuse swellings, with no visible lumps or areas of calcifiCation. She had total bilateral excision of the hypertrophied axillary breasts, and bilateral breast amputation with composite nipple-areola complex graft of the normally located breasts.The total weight of the breast tissues removed was 44.8 kilogram. CONCLUSION: Macromastia of this size is very rare. This case to date is probably the largest in the world literature. Surgical treatment of the condition gives a satisfactory outcome. PMID- 18399347 TI - Meigs' syndrome presenting as haemorrhagic pleural effusion. AB - BACKGROUND: The association of a benign ovarian tumor with ascites and hydrothorax that resolve after tumor resection, known as Meigs syndrome is a rare clinical entity. Rarer still is the haemorrhagic form of the syndrome OBJECTIVE: To describe a case of benign ovarian tumour associated with ascites and bloody pleural effusion. METHODS: A thirty-seven year old woman was referred for the further management of a pleural effusion. A detailed clinical evaluation was carried out, including pleural fluid cytology, chest CT scan and laparatomy. Treatment included antituberculous therapy and finally ovariectomy. RESULTS: The physical examination and a pelvic ultrasonographic scan revealed ascites in addition to a right sided ovarian mass. A chest CT-scan did not show any intrathoracic mass. Repeated pleural fluid cytology showed mesothelial cells but was negative for malignancy. An ovariectomy was performed and histological examination revealed a thecoma fibroma. The pleural effusion and ascites resolved spontaneously thus confirming the diagnosis of Meigs' syndrome. CONCLUSION: Meigs' syndrome should be considered in the differential diagnosis in female patients with hemorrhagic pleural effusion. PMID- 18399348 TI - Teaching undergraduate MBBS pharmacology in Nigeria: a need for curriculum modification. PMID- 18399349 TI - [Avian egg's white ovomucoid as food-allergen for human]. AB - Hen eggs are considered as the most common reason of a food allergy in humans. The most important allergens of egg white proteins are as follows: ovomucoid, lysozyme, ovalbumin and ovomucin. Ovomucoid is a Kazal-type protease inhibitor which accounts for about 10% of avian egg white protein. It is a glycoprotein containing 20 through 25% carbohydrates. The molecule of ovomucoid is composed of three homologous domains. All avian ovomucoid domains contain six cysteines in similar location that form three intradomain disulfide bonds. Ovomucoid (Gal d1) is one of the major allergen in hen's egg. It is a glycoprotein comprising 186 amino acids, and it has a molecular weight of 28000 Da and an isoelectric point of 4.1. Ovomucoid has antibacterial activity resulting from its ability to inhibit bacterial proteolytic enzymes crucial for microbial growth. Many studies reveal that ovomucoid is a thermo stable molecule. PMID- 18399351 TI - [New face of antiapoptotic proteins. I. Protein Mcl-1]. AB - Main regulators of apoptosis belong to Bcl-2 protein family and apoptosis inhibitory proteins--IAPs. In this review the apoptosis inhibitor--Mcl-1 protein is profoundly characterized. It is important that this unique short-living protein--the member of Bcl-2 family may also operate as apoptosis promoting agent, which results of alternative splicing of its pre-mRNA, posttranslational modifications or proteolysis. The review presents also other functions of Mcl-1, i.e. involvement in cell cycle regulation, elongation of telomers. Elevated expression of Mcl-1 accompanies the development of various cancers, neurodegenerative disorders and also infectious diseases. The obtained results indicate that expression level of Mcl-1 may be useful in treatment decisions of large number of diseases. Ablating expression of this protein may be an attractive therapeutic strategy in the treatment of various cancers, and the diseases where Mcl-1 may play a key role in apoptosis supression. PMID- 18399350 TI - [HAX-1 protein: multifunctional factor involved in apoptosis, cell migration, endocytosis and mRNA transport]. AB - HAX-1 protein, an anti-apoptotic factor, first identified in 1997, is also involved in cell migration, endocytosis and probably mRNA transport. HAX-1 structure indicates similarity to the proteins form Bcl-2 family, although there is no strong homology. HAX-1 is a substrate for Omi/HtrA2, a protease responsible for degradation of the caspases, and functions as an inhibitor of caspase-9, which points to its role in the regulation of apoptosis. Several HAX-1 interactions with proteins involved in apoptosis and cell motility were demonstrated. Another line of inquiry focus on its ability to bind 3' untranslated regions of the certain mRNAs. Some data indicate that it might be involved in mRNA transport. HAX-1 multifunctionality and its involvement in the processes important for the cell status suggest its possible role in oncogenesis and metastasis. It is also known that HAX-1 deficiency or overexpression leads to hereditary or systemic diseases (Kostmann disease, lesional psoriasis, systemic sclerosis). Therefore, detailed analysis of HAX-1 functions could be medically important. PMID- 18399352 TI - [New face of antiapoptotic proteins. II. Survivin]. AB - Survivin (mol.wt. 16.5 kDa; pI 5.1) belongs to the IAPs family--inhibitors of apoptosis. The human survivin protein contains an N-terminal BIR domain connected with a C-terminal, alpha-helical domain interacting with microtubules via a linker region. The BIR domain of the protein exhibits anti-apoptotic activity and plays a role in the binding of the chromosomal passenger complex (CPC) to the centromere regions of chromosomes. Alternative splicing of the human survivin gene gives rise to five different mRNA transcripts yielding wild-type survivin (142 aa) and four isoforms of the protein. In this review, the structure, features, and functions of wild-type survivin and its isoforms in the apoptotic process, cell cycle, and carcinogenesis as well as the significance of this protein as potential neoplastic marker are presented. Moreover, insights into the development of new anti-cancer therapeutic strategies targeting survivin are overviewed. PMID- 18399353 TI - [Regulation of gene expression by nitric oxide]. AB - During the past over 20 years, nitric oxide has become a subject of intensive research in molecular biology and other fields of science. Conducted studies discover its important role both in physiological and pathophysiological processes. A significant feature of nitric oxide is its participation in the regulation of gene expression. Mechanisms of this regulation mainly are based on direct action and consist in the modulation of transcription factors, of the translation and stability of mRNA, as well as in the posttranslational modification of primary gene product. The aim of this review is to present current knowledge about the influence of nitric oxide on the regulation of eukaryotic gene expression. PMID- 18399354 TI - [Nitric oxide production in plants]. AB - There are still many controversial observations and opinions on the cellular/subcellular localization and sources of endogenous nitric oxide synthesis in plant cells. NO can be produced in plants by non-enzymatic and enzymatic systems depending on plant species, organ or tissue as well as on physiological state of the plant and changing environmental conditions. The best documented reactions in plant that contribute to NO production are NO production from nitrite as a substrate by cytosolic (cNR) and membrane bound (PM-NR) nitrate reductases (NR), and NO production by several arginine-dependent nitric oxide synthase-like activities (NOS). The latest papers indicate that mitochondria are an important source of arginine- and nitrite-dependent NO production in plants. There are other potential enzymatic sources of NO in plants including xanthine oxidoreductase, peroxidase, cytochrome P450. PMID- 18399355 TI - [H(+)-coupled heavy metal transport in plants]. AB - It has been recently well documented that metal transport systems play a crucial role in the uptake, distribution and detoxification of heavy metals throughout the plant. A range of gene families that are likely to be involved in essential and non-essential metal transport has been now identified and their plasma membrane and/or tonoplast localization in plant cells has been recently confirmed. These include the primary metal transporters, using ATP as the source of energy and H(+)-coupling transporters, utilizing the electrochemical gradient previously generated by plasma membrane and tonoplast proton pumps. As the presence of nucleotide binding domains in the protein sequence may indicate its ATP-hydrolytic activity, it is more difficult to determine the H(+)-coupling activity of protein on the base of its structure. Thus, the H(+)-coupling activity of protein may be only proved by functional analysis of the protein. In this work, we briefly review the structure, regulation and function of the metal transporters operating as H(+)/metal cotransporters. PMID- 18399356 TI - [Alkaloids of Solanaceae (nightshade plants)]. AB - Alkaloids are nitrogen containing compounds found in many plants. They are products of plants secondary metabolism derived from amino acids, purines, pyrimidines or terpene. Most of them are drugs. The biological activity of some alkaloids has led to their intensive exploitation by humans, as pharmaceuticals, narcotics or poisons. During the past 30 years, major technical advances have led to substantial progress in our understanding of alkaloid biochemistry, but since then biosynthetic pathways of some alkaloids are not explained. The nightshade (Solanaceae) are widespread family of plants containing tropane alkaloids or glycoalkaloids. Both of them are naturally produced, as a defense mechanism against insects, predator and disease. On the other hand, most of the species of Solanaceae family have been used by human since several centuries. PMID- 18399357 TI - [5-n-alkylresorcinols of whole grain cereals and whole grain cereal products as biomarkers of healthy food]. AB - Epidemiological studies suggest that consumption of whole grain cereals and whole grain cereal products have many benefical health effects, including reducing risk of diabetes, obesity, coronary heart diseases, stroke and even some cancers. Precise knowledge protective compounds present in cereal grains can be achieved only when specific biomarkers (biological marker, indicator), that could provide estimation of grain cereals absorption and intake, are established and determined. 5-n-alkylresorcinols (main fraction of phenolic compounds in cereals), because of their specific occurrence only in bran fraction, obtained in refining of milling fractions process, could be a very good candidate to play the role of biomarker of whole grain intake. They are absorbed by animals and humans, present in human plasma and as metabolites in urine. Because composition of saturated homologues of 5-n-alkylresorcinols is different in rye and wheat grains, they could be used as an indicator of the intake of the specific type of cereals and whole grain cereal products. PMID- 18399358 TI - [The role of JNK pathway in familial Parkinson's disease]. AB - Parkinson's disease is a neurodegenerative disorder characterized by a dramatic loss of dopaminergic neurons in the substantia nigra. Among the many pathogenic mechanisms thought to contribute to the demise of these cells in sporadic cases of PD, oxidative stress has taken center stage due to extensive experimental evidence showing that dopamine- or MPTP-derived reactive oxygen species and oxidized dopamine metabolites may trigger toxicity through mitochondrial inhibition or deleterious modifications of biomolecules. In familial forms of PD, however, the involvement of toxic protein aggregation (synuclein), impairment of ubiquitin-proteosome system (parkin. and loss of antioxidative properties (DJ-1) has gained attention. Recently, JNK pathway has come to light that could link malfunction of mutated DJ-1, parkin, PINK1 and alpha-synuclein to the oxidative stress-triggered apoptosis, finally ascribing a common pathogenic mechanism to both the sporadic and familial forms of PD. PMID- 18399359 TI - Culturally competent care: a strategic imperative. PMID- 18399360 TI - Reducing health disparities through culturally sensitive treatment for HIV+ adults in Haiti. AB - There is a critical need to deliver empirically validated interventions to underserved populations. Haiti, the country most heavily affected by the AIDS epidemic in the Caribbean, accounts for approximately 50% of all cases in the region. Poverty, disparities in access to healthcare, and socio-political instability are among the reasons why the country has been ravaged by the disease. Ongoing projects in Haiti have shown that integrated prevention and care in resource poor settings are feasible and can be successful, as evidenced by a 50% drop in incidence among pregnant women since 1993. The AIDS prevention program has embarked on a comprehensive effort to culturally adapt a cognitive behavioral stress management program for Haitian HIV+ individuals. The purpose of the program is to improve adherence to antiretroviral medication, reduce transmission to uninfected partners, and improve coping. This comprehensive approach is necessary to ensure the validity of the cross-cultural adaptation of this intervention. PMID- 18399361 TI - Prostate cancer screening in African American and Caribbean males: detriment in delay. AB - Men of the African diaspora are diagnosed with prostate cancer much later than Caucasians and the mortality rate is significantly higher in these groups than among Caucasians. This study investigates health beliefs surrounding prostate health in a sample of African American and Caribbean men and identifies reasons men have for delaying or avoiding prostate screenings. One hundred African American and Caribbean men recruited from three churches, aged 37-89, were surveyed about their health seeking behaviors and knowledge of prostate cancer. Forty-five of these men also attended a seminar on the importance of early detection. Eighty percent of the men revealed they were embarrassed to have digital rectal examinations. Sixty percent feared impotence and incontinence after treatment if diagnosed with cancer. Findings reveal that attention to cultural realities may assist healthcare professionals in planning culturally sensitive educational interventions in the community that may narrow the health disparities gap in this population. PMID- 18399362 TI - Risk behaviours and healthcare needs of homeless drug users in Saint Lucia and Trinidad. AB - In the Caribbean AIDS has become the leading cause of death among those aged 15 45. The homeless are at high risk due to the nexus of mobility, psychiatric disorders and substance use, particularly crack. Seventy-four homeless, out-of treatment drug users were recruited in Saint Lucia and Trinidad to assess risk behaviour and healthcare needs. Information was collected in several domains: medical, mental health, substance use, sexual risk, and barriers to care. Ninety five percent of the sample reported crack cocaine use within the previous 30 days. Approximately 35% of the sample had ever been diagnosed with a sexually transmitted disease and 25% of the sample reported being HIV+. Approximately 40% had a history of trading sex for crack or money. Due to the range of needs in this population, interventions should be multimodal. A focus on the individual, community, and structural levels is warranted to improve chances of program effectiveness. PMID- 18399363 TI - Increasing access to preventive health care through cultural adaptation of effective HIV prevention interventions: a brief report from the HIV prevention in Haitian youths study. AB - This article describes an HIV prevention study among Haitian youths, based on the cultural adaptation of a cognitive behavioral HIV risk reduction intervention entitled "Becoming a Responsible Teen." The aim of the parent study is to evaluate whether the BART intervention is more effective than a control condition in reducing HIV risk behavior in the target population. The project explores how self-efficacy, behavioral intentions, social factors and acculturation influence the risk behavior of Haitian American adolescents. This community based translation of a risk reduction intervention, previously found to be effective in other populations, can serve as a model for reducing health disparities in a vulnerable adolescent population that lacks access to preventive health care. The purpose of this paper is to present preliminary data from the pre-assessment phase of the intervention, which enabled the project to incorporate health disparity issues with this population and address barriers to health care access. PMID- 18399364 TI - A quantum shift in home care service delivery. PMID- 18399365 TI - Long term urinary catheter problems: a flow chart to aid management. AB - In this article the author presents a flow chart designed to assist nurses in providing evidence-based care of long term urinary catheters. The flow chart guides the nurse in making a proper assessment of the reason for urinary bypassing, blockage and non-drainage. The flow chart is intended to help identify a cause and to aid the practitioner in following a long term management plan. PMID- 18399366 TI - Review of personal protection equipment used in practice. AB - Standard precautions offer a consistent approach to infection control that does not rely on knowledge or suspicion of infection, and contributes to staff and patient safety by reducing the risk of exposure to potentially infectious material. One of the cornerstones of standard precautions is the appropriate use of personal protective equipment (PPE) whenever contact with blood or body fluids is anticipated. However, evidence suggests that compliance with standard precautions including correct use of PPE is inadequate. Demographic and epidemiological changes in the U.K., and the drive to provide more complex patient care outside acute hospitals may lead to increased infection risks for both patients and community healthcare workers. This review examines the importance and use of PPE by community nurses and discusses the implications for practice of poor compliance with standard precautions. Recommendations for practice will be made aimed at improving compliance with this important element of standard precautions. PMID- 18399367 TI - Dealing sensitively with sexuality in a palliative care context. AB - Sexuality is integral to a person's identity and is encompassed in holistic palliative care. Current directives emphasize psychosocial support that is sensitive to a variety of issues, including sexuality, and offer community nurses the mandate to facilitate expressing sexuality. Nurses fail to address sexuality for a variety of reasons. Appreciating patients' individuality and avoiding making assumptions are some of the requirements to enabling expression of sexuality. Nurses need wisdom and determination to discuss such issues, and it is important that they recognize opportunities and cues as they arise. This article provides an examination of the issues surrounding sexuality outlining an approach that captures' constant attention to details. This guides the 'what', 'how' and 'why' sexuality in palliative care can be addressed. Ethics and safety aspects are briefly discussed and implications for practice suggested. Educating staff and focused education for the patient can positively influence sexuality and individuals' quality of life. PMID- 18399368 TI - The evolving role of the community nurse specialist in palliative care. AB - The community clinical nurse specialist in palliative care role has evolved in an uncoordinated manner resulting in a variety of grades of nurse with differing role expectations. In general the CNS role consists of a clinical, educational, strategic and managerial remit. The reality of working with such a diverse role creates tension and conflict for the CNS between personal, organization and practical expectations. Despite this the community palliative care CNS is central to the future development of palliative care services in the community generating a culture of sharing knowledge and expertise. This will however require further investigation into the impact these demands will have on the individual. PMID- 18399369 TI - Survey to investigate the role of the community stroke care coordinator. AB - The National Service Framework for older people recommends that patients recovering from a stroke should have access to a stroke care coordinator (SCC) to support their longer-term recovery. However, the knowledge base, skills and procedures for the role are poorly defined. We have therefore conducted a survey to examine the development and responsibilities of SCCs with a remit for longer term support. We identified 39 SCCs, most of whom were nurses. Three role types were observed: enhanced community follow-up (n=22); hybrid (n=15); and hospital outpatient (n=2). However, there was overlap between the roles reflecting a wide variation in practice in respect of timing and frequency of patient contacts, and methods for assessing patients' needs. Workload and knowledge of the evidence base were identified as the main challenges to delivering care. Meeting patients' needs was impeded by limited access to psychology and physical therapies. A more robust evidence-base to support longer-term stroke care, combined with routine clinical governance systems for monitoring care provision could enhance the development of the SCC role. PMID- 18399370 TI - Modernising your nursing career. PMID- 18399371 TI - The new national framework for NHS funded continuing care. AB - This article outlines the new National framework for NHS funded continuing care and NHS funded nursing care (Department of Health, 2007) and the implications for community nurses. Successful implementation is dependent upon good multidisciplinary assessment and person-centred care planning. Good practice in these areas is described. The role of the registered nurse in carrying out an assessment for NHS funded continuing care, the importance of record keeping and professional accountability are discussed. PMID- 18399373 TI - Doing well but room for improvement. PMID- 18399372 TI - Legal requirements for conducting clinical trials. AB - Clinical trials are crucial to the development of new medicines as they test the safety and effectiveness of the drug on human subjects. However, trials do pose a risk to the participants. To minimize this risk all clinical trials must meet the requirements of the Clinical Trials Directive (Council Directive (EC) 2001/20 2001) and the Medicines for Human Use (Clinical Trials) Regulations 2004. In this article Richard Griffith discusses the legal framework that regulates clinical trials in the United Kingdom. PMID- 18399374 TI - What are the new Trauma Codes, Form Locator (FL) 19, Type 5 and Revenue Code, 68x "G" 0390 and APC 0618 and how should they be implemented? PMID- 18399375 TI - Trauma 2007: a finger on the pulse. AB - Truma surgery today is facing a number of significant challenges that offer a stimulus for growth and evolution of tl practice. To successfully face these challenges, reexamination of the discipline, the current practice models for its providers, and the definition/scope of the specialty will be necessary. Further development and application of the cute care surgery model may represent the future direction for trauma care practitioners. PMID- 18399376 TI - Perceptions of nurses' caring behaviors by trauma patients. AB - A descriptive study of moderately to severely injured patients' perceptions of nurses' caring behaviors was conducted at a level I trauma center. Seventy patients older than 18 years admitted through the trauma resuscitation unit were included. The participants were interviewed in their rooms/cubicles by the research team to complete the Caring Behaviors Inventory. The average rating for the sample was 4.8. Patients receiving care through trauma services expressed an overall positive perception of caring from the nursing staff while there was room for improvement in some areas. PMID- 18399377 TI - The pupillary response in traumatic brain injury: a guide for trauma nurses. AB - Trarumatic brain injuries (TBIs) affect more than 1.4 million Americans annually. Trauma nurses caring for these patients routinely perform serial neurologic assessments, including pupillary examinations. While trauma nurses are likely familiar with basic components of the pupillary examination, some confusion about more specific aspects of the examination and the physiologic basis of the pupillary response may still remain, particularly as it pertains to patients with TBI. Therefore, the purpose of this article is to identify the key components of a pupillary examination and its associated physiologic response. A case study is provided to illustrate the application of this information among patients with TBI. PMID- 18399378 TI - Pediatric trauma nursing education requirements: a national overview. AB - This study describes current trauma nursing education requirements and nursing perception for additional pediatric trauma education. A web-based survey was electronically distributed to members of Society of Trauma Nurses. Overall, a lack of consistent standards across the United States for what constitutes pediatric trauma education was noted. Many hospital trauma programs expend time and money developing their own hospital course. Strong support exists for the development of an additional pediatric trauma course with a skills station. Basic concepts of primary/secondary survey, airway management, and fluid management for hypovolemic shock should be a high priority within this curriculum. PMID- 18399379 TI - Young riders: all-terrain vehicle use in north Texas. AB - Annually, there are more than 54,000 visits to nationwide emergency departments as a result of injuries sustained while riding all-terrain vehicles. Of those visits, nearly one-half involve children younger than 16 years. Children's Medical Center Dallas admitted 211 patients over a 5-year period from March 2002 until May 2007 as a result of all-terrain vehicle-related incidents with 207 of those patients aged 13 years or younger. The primary mechanisms of these injuries were secondary to collision with stationary objects, rolling of the vehicles, and falling from the vehicles. PMID- 18399380 TI - The growth and development of a level II trauma center. AB - Attaining verification as a Level II Trauma Center requires dedication, flexibility, and continuous education. This article contains the history, birth, and growth of a Level II Trauma Center through a trauma resource clinician's experiences. It is intended to share the thoughts, processes, and technological advances of establishing a Level II Trauma Center. PMID- 18399381 TI - An overview of New Zealand's trauma system. AB - Patterns of trauma and trauma systems in New Zealand are similar to those in Australia. Both countries have geographical considerations, terrain and distance, that can cause delay to definitive care. There are only 7 hospitals in New Zealand that currently manage major trauma patients, and consequently, trauma patients are often hospitalized some distance from their homes. The prehospital services are provided by one major provider throughout the country, with a high level of volunteers providing these services in the rural areas. New Zealand has a national no-fault accident insurance system, the Accident Compensation Corporation, which funds all trauma-related healthcare from the roadside to rehabilitation. This insurance system provides 24-hour no-fault personal injury insurance coverage. The Accident Compensation Corporation provides bulk funding to hospitals for resources to manage the care of trauma patients. Case managers are assigned for major trauma patients. This national system also has a rehabilitation focus. The actual funds are managed by the hospitals, and this allows hospital staff to provide optimum care for trauma patients. New Zealand works closely with Australia in the development of a national trauma registry, research, and education in trauma care for patients in Australasia (the islands of the southern Pacific Ocean, including Australia, New Zealand, and New Guinea). PMID- 18399382 TI - Management of venous thromboembolism in the palliative care setting. AB - Recent literature has highlighted the unique challenges facing professionals involved with the management of venous thromboembolism (VTE) in cancer patients. Although research favours the use of low molecular weight heparin (LMWH) rather than warfarin, no studies have been conducted exclusively in the palliative care population. Likewise, active treatment may not always be appropriate in the advanced cancer patient. This paper highlights the issues to be considered when facing a palliative care patient and suspected VTE. In particular, consideration must be given to the practicalities and acceptability of investigating suspected VTE, the dose and length of anticoagulation, and management of the patient nearing death. An understanding of the current evidence must be applied to the individual patient, taking into account their needs and wishes. PMID- 18399383 TI - The paradox of palliative care nursing across cultural boundaries. AB - The purpose of this qualitative study, using a modified grounded theory approach, was to investigate how palliative care nurses care for people from cultural backgrounds other than their own. Ten palliative care nurses were interviewed. The semi-structured interviews were tape-recorded and transcribed verbatim. Analysis commenced as soon as data began to emerge. The findings show that palliative nursing across cultural boundaries is paradoxical. In endeavouring to treat everyone equally, nurses treated everyone as individuals. They made intense efforts to transcend both cultural and language barriers. The nurses gave of themselves when caring for the patients, but their endeavours were impeded by limited resources and a lack of education. The theory of cross-cultural endeavour in palliative nursing was developed to explain how palliative care nurses care for patients from cultures other than their own. PMID- 18399384 TI - Children's needs when facing the death of a parent from cancer: part two. AB - A small exploratory study was conducted to identify the psychosocial needs of children facing the death and subsequent bereavement of a parent from cancer. The focus was on the palliative phase of care through the bereavement period with the aim of identifying the best way forward in further developing a community-based service for children in Fife. Part one selectively reviewed the literature related to children's and families' needs when facing and dealing with the death of a parent, and described. Part two presents the results and reports the issues which will be considered as a basis for future service development. PMID- 18399385 TI - Macmillan nurse facilitators: conducting a training needs assessment for district nurses. AB - This article will describe how Macmillan nurse facilitators conducted a training needs analysis in order to inform and develop an education programme in palliative care for district nurses. A questionnaire was developed and distributed to all 250 district nursing staff in the north/east sector of Glasgow to identify the priority training issues relating to palliative care. This included all district nursing sisters, first and second level registered nurses and auxiliary nursing staff working within the community setting. An 85% response rate to the questionnaire was achieved and an education programme was developed and delivered to address the needs identified by staff. This article goes on to describe the process of the training needs analysis and the benefits of providing an education programme that addresses the educational needs of staff members. Reference is made to a larger evaluation study which highlights staff members' views on both these points. PMID- 18399386 TI - Authentic performance appraisal: when home is the workplace. AB - Home care services have characteristics that are distinctive from other health care settings. Thus, they require different management systems to those traditionally used in institutional settings. Typical management models often require adaptation for home care services in areas such as staffing, education and support. This includes performance appraisal, where traditional models may be inappropriate when clinicians work alone, with little supervision. This article describes an experience of adapting a performance appraisal tool, in order to be more relevant for staff working in home-based care. The project arose from staff frustration with the expectation of management that a tool developed for institutional settings would be appropriate for the home setting. This frustration gave the impetus for developing a successful appraisal system, and because of staff involvement, the model was wholeheartedly embraced. The process of development is discussed, and the model explained, with commentary about its implementation in a home care service. PMID- 18399387 TI - Use of a modified ESAS in cancer patients: a pilot study of patient and staff experiences. AB - AIM: To evaluate the feasibility of a modified Edmonton Symptom Assessment Scale (ESAS) for monitoring symptoms in oncological palliative care. METHODS: A modified ESAS was delivered daily to 28 patients with advanced cancer. A questionnaire to discover staff members' opinions on the ESAS was delivered at the end of the study. Structured interviews were used to examine patients' opinions on the ESAS. RESULTS: The mean total ESAS score was 28.9 on inclusion day and 25.8 on Day 3 (p=0.531). Eleven of 21 of the staff considered the ESAS to be a 'good' or 'very good' way to obtain information about patients' symptoms. Seventeen of 24 patients who participated in the interview felt that ESAS was easy to fill in, and that there were no missing questions. CONCLUSION: The patients felt that the modified ESAS contained relevant symptoms, and that the questionnaires were easy to fill in. The staff considered the modified ESAS to be a useful instrument for obtaining information about patients' symptom distress. The modified ESAS is a good instrument for use as part of the daily clinical routine, as well as for monitoring symptoms in palliative oncological care. PMID- 18399388 TI - The waiting game we cannot afford to lose. PMID- 18399389 TI - Incompetence and empty pockets: all in a day's work. PMID- 18399390 TI - Unpacking the question of graduation. PMID- 18399391 TI - Women's help-seeking behaviour at the onset of myocardial infarction. AB - Few studies have explored the factors affecting women's decisions to seek medical assistance at the onset of cardiac symptoms. Although some studies have shown that there are gender differences between men and women in terms of how they cope with having a myocardial infarction (MI) and heart disease, few have explored how women actually experience MI and what influences their help-seeking behaviour. Those studies that have, suggest that women often fail to recognize the signs of MI and delay seeking treatment for symptoms of MI longer than men do. This qualitative study explores the female experience of MI, focusing on some of the thought processes and coping strategies used by women when having an MI. Using a grounded theory research approach, 25 post-MI female patients were interviewed about their experiences during acute MI. Through analysing and coding data, a number of categories emerged that provide insight into the health seeking behaviour of women at the onset of MI. Emergent categories were: a delay in seeking help (main category); an engendered perception of heart disease (sub category); not recognizing symptoms (sub-category); maintaining control through self-medication (sub-category). Analysis of the data suggests that when experiencing an MI, women delay seeking help for a number of reasons, including not recognizing symptoms, perceiving heart disease as a 'male' problem and preferring to self-medicate. PMID- 18399392 TI - The Sepsis Six: helping patients to survive sepsis. AB - Sepsis kills more people than lung cancer, and more people than bowel and breast cancer put together. The costs to the NHS are significant; it is estimated that in Europe, patients with severe sepsis cost healthcare funders around 7.6 billion euros per year (Daniels et al, 2007). Costs in the United States are estimated at $16 billion annually (Angus et al, 2001), and in the United Kingdom up to 46% of intensive care unit (ICU) bed days are used by patients with severe sepsis (Padkin et al, 2003), with each ICU bed costing around pounds sterling1700 per day. In 2002 an international campaign was launched: the Surviving Sepsis Campaign. The main aim of this campaign is to reduce mortality from sepsis by 25% by 2009. A lot of the early work has concentrated on improving sepsis care in intensive care units, but many patients on general wards develop sepsis, and the need to educate nurses throughout all areas of the hospital has been recognized. In September 2007 a new part of the campaign was launched called Survive Sepsis, which aims to deliver sepsis education to ward nurses and junior doctors. This article discusses how to recognize severe sepsis and explains how nurses can dramatically improve a patient's chance of survival by ensuring that six simple things (Sepsis Six) are done in the first hour. PMID- 18399393 TI - Care or harm: exploring essential components in skin care regimens. AB - Skin care is a fundamental aspect of basic nursing care, with the outcome of these interventions often used to gauge the quality of the care provided. However, possibly as a result of the 'basic' nature of this aspect of care, little attention has been paid to developing a firm evidence base to guide decision-making in this area. This is compounded by the wide variety of products now available, ranging from simple cleansers and moisturizers, to more complex agents that cleanse, moisturize and protect all in one, which adds to the confusion. This article explores the basic principles of skin care, and considers the evidence available to support the nursing interventions used, and the range of products available. Finally it suggests the adoption of clear skin care protocols, and recommends actions that should form part of these. PMID- 18399394 TI - The use of antiembolic stockings. part 2: a clinical audit. AB - AIM: To audit current practice regarding selection and use of graduated elastic compression stockings (GECS) in the authors' Trust. Following a literature review it was important for the Trust that the authors audited current practice before the development and implementation of any new guidelines. BACKGROUND: A literature review enabled the authors to establish best practice principles for the use of GECS and develop guidelines for use across their Trust. However, feedback from various sources highlighted conflicting practices regarding the methods by which patients either did or did not receive stockings, and which lengths and brands were used. As the guidelines aimed to allow implementation of standardized change across the Trust, it was important to establish current practice with regard to GECS selection and use. Therefore, an audit tool was developed carried out on the use of GECS. METHODS: An audit tool was developed and implemented to establish the practices before the implementation of the guidelines. RESULTS: The audit demonstrated that there was no consistent policy within the surgical services directorate for the correct use of GECS. CONCLUSIONS: It is important that all healthcare providers have a local policy for GECS use, which makes it clear how an assessment for patients requiring GECS needs to be conducted and how that assessment is documented. PMID- 18399395 TI - Undertaking a literature review: a step-by-step approach. AB - Nowadays, most nurses, pre- and post-qualification, will be required to undertake a literature review at some point, either as part of a course of study, as a key step in the research process, or as part of clinical practice development or policy. For student nurses and novice researchers it is often seen as a difficult undertaking. It demands a complex range of skills, such as learning how to define topics for exploration, acquiring skills of literature searching and retrieval, developing the ability to analyse and synthesize data as well as becoming adept at writing and reporting, often within a limited time scale. The purpose of this article is to present a step-by-step guide to facilitate understanding by presenting the critical elements of the literature review process. While reference is made to different types of literature reviews, the focus is on the traditional or narrative review that is undertaken, usually either as an academic assignment or part of the research process. PMID- 18399396 TI - The Mental Capacity Act 2005 and decision-making: advance decisions. AB - One of the crucial features of the Mental Capacity Act (MCA) 2005 was recognizing in a statute the ability of a person to refuse treatment at a subsequent time when he or she lacked the mental capacity to decide on treatment. This refusal, known as an advance decision or living will or advance refusal had been recognized at common law, i.e. judge-made law. This article, continuing the series on the MCA explains the statutory provisions, the specific requirements which must be satisfied if life-sustaining treatment is refused in advance and looks at a practical situation to illustrate the statutory provisions. The link between advance decisions are lasting powers of attorney is also discussed. PMID- 18399397 TI - Pain management for the opioid-dependent patient. AB - This article describes the development, introduction and implementation of'Guidelines for Pain Management in the Opioid-Dependent Patient' in Yeovil District Hospital NHS Foundation Trust, Somerset. It incorporates the relevant background experiences within the organization and establishes the rationale for the development of the guidelines. An outline of the consultation processes with the different professional groups involved is given, together with the strategy for implementation across the Trust. This project challenged old ways of working and embraced new expanding roles. Indications are that both the quality of care and the patient experience have improved as a direct result of the implementation of these guidelines. PMID- 18399398 TI - Nursing interventions for preventing alcohol-related harm. AB - Harrington-Dobinson and Blows recently provided a three-part series of articles on alcohol, its consequences for health and wellbeing, and the role of the nurse. Their third article outlined the health education and health promotion role of the nurse. They outlined basic principles for nursing practice in relation to the patient with alcohol dependence in the acute general hospital. The authors of this article believe that much more can, and must, be said in relation to the vital issue of nurses' clinical interventions for alcohol. This article builds on the third article from Harrington-Dobinson and Blows by outlining, in more concrete terms, how nurses in all settings can effectively intervene with patients. It introduces the current evidence-based guidelines in this area and use the 'consensus model' contained within them to describe the process of effective alcohol intervention. Using dialogue examples to illustrate the research, the authors introduce the literature on brief interventions and motivational interviewing to the nursing audience. PMID- 18399399 TI - Nursing electives: an innovative and creative learning opportunity. AB - Nursing elective can provide the nurse (learner or registered nurse) with a host of new opportunities to experience and observe practice from a variety of different perspectives. The nursing elective should be planned well in advance and much time and thought should be given to preparing for this activity. There are a number of resources (human and material) available to help with the planning of the event and as such careful and thorough planning at an early stage can make the activity much more successful. The funding of the elective event needs to be thought about early on in the planning stage if this is likely to be an issue. Practical considerations, such as insurance cover, honorary contracts, travel and other documents must also be considered carefully. The elective opportunity has the potential to provide the individual with a unique experience, a chance to delve deeper in relation to a particular area of interest as well as potentially experiencing the provision of health care through an alternative lens. The success of the elective largely depends upon the amount of effort the individual puts into it. PMID- 18399400 TI - Time in the professional gym. PMID- 18399401 TI - Strapped and other topographically nonplanar calixpyrrole analogues. Improved anion receptors. AB - Calixpyrroles and related macrocycles are non-aromatic synthetic anion receptors that have attracted considerable attention in recent years. The unfunctionalized, parent calix[4]pyrrole system, also known as octamethylporphyrinogen, may be prepared in one step and in high yield from pyrrole and acetone, and is an effective anion receptor, showing a preference for fluoride, phosphate, carboxylate and chloride anions in organic media. Efforts to improve the anion binding affinity of calix[4]pyrrole and to modify its inherent selectivity have led to the synthesis of a variety of new, modified calixpyrroles. Among the most effective of these are derivatives that contain bridging "straps". In this Feature Article, the preparation and properties of these and other topographically nonplanar calixpyrrole analogues are reviewed from the perspective of the anion recognition chemist. PMID- 18399402 TI - Trichlorogallium and trialkylgalliums in organic synthesis. AB - Organic gallium compounds formed by the interactions of organic compounds with trichlorogallium or trialkylgalliums exhibit various reactivities, and their use in organic synthesis is described. PMID- 18399403 TI - Can large magnetic anisotropy and high spin really coexist? AB - This theoretical study discusses the interplay of the magnetic anisotropy and magnetic exchange interaction of two Mn6 complexes and suggests that large magnetic anisotropy is not favoured by a high spin state of the ground state. PMID- 18399404 TI - Direct observation of change in the molecular structure of benzyl (Z,Z)-muconate during photoisomerization in the solid state. AB - For the solid-state photoisomerization of benzyl (Z,Z)-muconate to the corresponding (E,E)-muconate, the direct observation of a change in the crystal structure has revealed that the isomerization occurs by a topochemical reaction process according to a bicycle-pedal model and is finally accompanied by a phase transition to a stable crystal structure. PMID- 18399405 TI - Spontaneous resolution of a 3D chiral polyoxometalate-based polythreaded framework consisting of an achiral ligand. AB - Two enantiomerically 3D chiral POM-based architectures have been constructed based on the achiral ligand bbi, [V10O26](4-) polyoxoanion and mixed valence Cu(I/II) without a chiral auxiliary, and they represent the first examples of enantiomerically 3D POM-based compounds using achiral ligands. PMID- 18399406 TI - Assembling metals and clusters around an octaphosphine ligand based on N substituted bis(diphenylphosphanyl)amines: structural characterization of dendrimer-like Co12 and Co16 branched clusters. AB - A polypodal ligand based on an aromatic ring decorated by four dppa-type diphosphine moieties has been used to prepare Pt(II) complexes and to assemble four tricobalt or tetracobalt carbonyl clusters, leading to centrosymmetric Co12 and Co16 "clusters of clusters" which were characterized by X-ray diffraction. PMID- 18399407 TI - High performance platinized titanium nitride catalyst for methanol oxidation. AB - Platinized TiN is shown to be a very good catalyst for the electrochemical oxidation of methanol; it does not contain Ru and the CO poisoning effect is alleviated by the presence of TiN. PMID- 18399408 TI - Preparation and photosensitizing property of novel Cd10S16 molecular cluster dendrimer. AB - A new Cd10S16 molecular cluster dendrimer has been prepared and characterized; photooxygenation reaction using the molecular cluster dendrimer as a photosensitizer was successful. PMID- 18399409 TI - Questioning the paradigm of metal complex promoted phosphodiester hydrolysis: [Mo7O24](6-) polyoxometalate cluster as an unlikely catalyst for the hydrolysis of a DNA model substrate. AB - The first example of a phosphodiester bond cleavage promoted by a highly negatively charged polyoxometalate cluster has been discovered: the hydrolysis of the phosphodiester bond in a DNA model substrate bis(p-nitrophenyl)phosphate (BNPP) is promoted by the heptamolybdate anion [Mo7O24](6-) with rates which represent an acceleration of nearly four orders of magnitude compared to the uncatalyzed cleavage. PMID- 18399410 TI - Intermolecular radical addition reactions of alpha-iodo cycloalkenones and a synthetic study of the formal synthesis of enantiopure fawcettimine. AB - The generation of alpha-carbonyl vinyl radicals from alpha-iodo cycloalkenones, the scope of their participation in intermolecular addition reactions with electron-withdrawing olefins are studied and a synthetic study of the formal synthesis of enantiopure fawcettimine using this reaction is described. PMID- 18399411 TI - Iron(III) amine-bis(phenolate) complexes as catalysts for the coupling of alkyl halides with aryl Grignard reagents. AB - Catalytic cross-coupling of aryl Grignard reagents with primary and secondary alkyl halides bearing beta-hydrogens is achieved using Fe(III) amine bis(phenolate) halide complexes. PMID- 18399412 TI - The barrier to enantiomerization of N-Boc-2-lithiopyrrolidine: the effect of chiral and achiral diamines. AB - (-)-Sparteine and TMEDA dramatically lower both enthalpy and entropy of activation for the barrier to enantiomerization of N-Boc-2-lithiopyrrolidine in diethyl ether, whereas N,N'-diisopropylbispidine has little effect; the entropy of activation for enantiomerization is zero in the presence of TMEDA and slightly negative in the presence of sparteine; these data suggest a subtle change in mechanism of enantiomerization in the presence of TMEDA and sparteine. PMID- 18399413 TI - Rh-catalyzed P-P bond activation. AB - A Rh-catalyst derived from (NacNac)Rh(COE)(N2) effects the hydrogenation and silylation of P-P bonds to give secondary phosphines and silylphosphines, (Ph2PH) and (Ph2PSiRR'2) respectively; the latter process is shown to also involve the silylation of secondary phosphines. PMID- 18399414 TI - Endocyclic extension of porphyrin pi-system in etheno-bridged N-confused tetraphenylporphyrin. AB - Etheno-bridged N-confused tetraphenylporphyrin was synthesized through flipping of the confused pyrrole ring and endocyclic extension of [18]annulenic pi conjugated system was exemplified by the absorption spectra as well as the calculated Kohn-Sham orbitals. PMID- 18399415 TI - 1,2-Addition of trialkylaluminium reagents to N-diphenylphosphinoylketimines in the absence of any additional reagents. AB - Using reactive trialkylaluminium reagents for the 1,2-addition on acetophenone- and benzophenone-derived ketimines, alpha-trisubstituted amines were obtained in excellent yields up to 99%. PMID- 18399416 TI - Metal-specific allosteric activation and deactivation of a diamine. AB - The reaction of a potentially tetradentate bis(pyridyl-thiazole) ligand with acetone is allosterically activated upon complexation with Cd(II) but deactivated by reaction with Cu(I), demonstrating metal-specific allosteric controlled reactivity. PMID- 18399417 TI - Anti-Markovnikov hydroamination and hydrothiolation of electron-deficient vinylarenes catalyzed by well-defined monomeric copper(I) amido and thiolate complexes. AB - Monomeric Cu(I) amido and thiolate complexes that are supported by the N heterocyclic carbene ligand 1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene (IPr) catalyze the hydroamination and hydrothiolation of electron-deficient vinylarenes with reactivity patterns that are consistent with an intermolecular nucleophilic addition of the amido/thiolate ligand of (IPr)Cu(XR) (X = NH or S; R = Ph, CH2Ph) to free vinylarene. PMID- 18399418 TI - Tagging alcohols with cyclic carbonate: a versatile equivalent of (meth)acrylate for ring-opening polymerization. AB - Cyclic carbonate monomers based on a single biocompatible scaffold allow for incorporation of a wide range of functional groups into macromolecules via ring opening polymerization. PMID- 18399419 TI - An organic p-type dopant with high thermal stability for an organic semiconductor. AB - To overcome the thermal instability of a p-doped organic hole transporting layer using the state-of-the-art p-type dopant, 2,3,5,6-tetrafluoro-7,7,8,8 tetracyanoquinodimethane, a potent electron accepter, 3,6-difluoro-2,5,7,7,8,8 hexacyanoquinodimethane, has been found to possess superior thermal stability and proved to be an excellent p-type dopant. PMID- 18399420 TI - Application of furyl-stabilized sulfur ylides to a concise synthesis of 8a-epi swainsonine. AB - The total synthesis of 8a-epi-swainsonine has been achieved in 20% overall yield from R-glyceraldehyde dimethylacetonide 3 through epoxidation with the achiral furyl-substituted sulfonium ylide 2d as one of the key steps. PMID- 18399421 TI - Diabrotica virgifera virgifera LeConte: inconspicuous leaf beetle--formidable challenges to agriculture. AB - In the universe of entomology with its close to one million described and an estimated ten million undescribed species, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysometidae), the Western corn rootworm (WCR), is one of the economically most important pests. It causes annual treatment costs and damages of a billion dollars in the US (Metcalf 1986). Similar costs are predicted for Europe which the beetle invaded 15 years ago. Due to lack of natural enemies it is now expanding its territory at a rapid rate. With prior experience gained in Illinois, USA, and subsequent largely unsuccessful efforts to stop the pest in Europe, eradication efforts, although attempted in France and in the Veneto region of northern Italy, are not a viable sustainable strategy for the future nor a long term solution. Crop rotation, so far one of the best cultural management options of practicing entomologists, is beginning to show weaknesses specificly on Glycine max (Shaw et al. 1978, Levine et al. 2002, Spencer et al. 2005, Kiss et al. 2005, Tollefson and Prasifka 2006). WCR also was discovered on alternative hosts such as Cucurbita pepo in Slovenia (Hummel et al. 2007a, 2007b), C.maxima in Hungary (Moeser and Vidal 2001), and Serbia (Baca 1993, Baca and Berger 1994, Baca unpublished 1998, Baca 2007). WCR thus turns out to remain a challenge to general entomolog'sts, agricultural and ecological entomologists, geneticists, epidemiologists, crop protection engineers, phytosanitary services and economists alike. WCR and H. sapiens today are reaching a labile ecological equilibrium of coexistence, with new surprises on both sides in the "arms race" just waiting around the corner. Most experts will agree that WCR is in Europe to stay and is readily expanding from here to Asia and Africa if given the slightest chance.- The really unsolved question remains how to manage WCR sensibly, economically, and sustainably, and how and where to train a new generation of capable, reliable, all round entomological pest managers needed for the future. PMID- 18399422 TI - Soy polyol formulations as novel seed treatments for the management of soil-borne diseases of soybean. AB - Polyurethanes prepared from vegetable oils display a number of desirable properties useful for many commercial and industrial applications. One unique application is that of an agricultural seed treatment. Seed treatments are used to incorporate pesticides onto the seed coat and to decrease the disease susceptibility of the seed during its germination in the soil. In addition, by altering the movement of water across the seed coat and by incorporating protective pesticides in the coating, seed coating polymers can enhance the germination and survival of the seed under adverse environmental conditions. Soy polyols alone, and in combination with glycerin, polymerized with 4,4' diphenylmethane diisocyanate (MDI) were studied for their seed treating properties and impact on soybean seed germination. The cross-linking density and properties of these polyurethane compounds were varied by changing the isocyanate/hydroxyl molar ratios. In order to optimize the coating qualities and to increase the efficiency of the coating, acetone was also studied as a diluting solvent to reduce the viscosity of the polyurethane mixture prior to polymerization on the seed coat. Optimal polymerization and resulting germination (95%) were obtained using a 1:1 isocyanate/hydroxyl molar ratio consisting of a mixture of soy polyol 180 and glycerin, and the use of an equal volume of acetone as a dilution solvent. This optimal polyurethane seed treatment had several desirable qualities including: reduced viscosity, decreased seed coating thickness, increased seed coating uniformity and permitted larger volumes of seed to be treated with the same volume of polymer. This optimal seed treatment increased the soybean seed germination by 15%, as compared with untreated seed. In addition, preliminary studies of the compatibility of these unique formulations with commercial and experimental fungicides also support the use of these polymers as seed treatments due to their enhanced stability, longevity and slow active ingredient water teaching characteristics. Compatibility of these seed coating polymers as formulations with captan, metalaxyl, thiabendazole and novel antimicrobial lipids and triterpenoid compounds display that the active ingredients can readily provide a zone of fungal inhibition around the seed as it germinates in the presence of Macrophomina phaseolino, causal agent of charcoal rot of soybeans. However, the release of the active ingredient from the polyol seed treatments is less affected by water leaching as compared to commercially available water-soluble seed treating polymer formulations. This is most likely due to the polyols unique polymer cross-linking characteristics. These results support the continued exploration of soy polyol derived polymers as seed coating compounds. PMID- 18399423 TI - Phototransformation of pesticides on plant leaves: the case of sulcotrione. AB - Photochemistry is one of the main ways of pollutants degradation in the environment. There is an obvious lack of data concerning the photostability of agrochemicals on plant foliage. We report here the first photodegradation study of a triketonic herbicide (sulcotrione) used as a substitute of atrazine. To mimic surface Leaves, we used films made of carnauba grey wax or of cuticular wax extracted from maize Leaves. Under simulated solar light, sulcotrione was rapidly phototyzed. The phototysis was much faster on cuticular wax films than in aqueous phase. Moreover, the formulated sulcotrione (Mikado) disappeared even more quickly than the pure active ingredient. Sulcotrione mainly underwent an intramolecular cyclization. To validate these observations, sulcotrione and its main photoproduct were monitored after maize field treatment. The day after the treatment the main photoproduct was detected on the maize leaves and suLcotrione had already significantly disappeared. This experiment demonstrates that phototysis can be a relevant process in real conditions after field treatment. PMID- 18399424 TI - The efficiency of adjuvants combined with flupyrsulfuron-methyl plus metsulfuron methyl (Lexus XPE) on weed control. AB - This paper presents the results of laboratory tests on a selection of weeds (Viola arvensis, Polygonum persicaria, Chamomilla recutita, Chenopodium album, Veronica persicaria, Alopecurus myosusroides) to investigate the efficiency of flupyrsulfuron-methyl plus metsutfuronmethyl (Lexus XPE) in combination with different adjuvants. The efficiency of the herbicide improved in combination of adjuvants. The level of phytotoxicity of the adjuvants-herbicide treatments appllied varied among the different weed species. PMID- 18399425 TI - Combinations of reduced rates of 1,3-dichloropropene and dazomet as a broad spectrum soil fumigation strategy in view of methyl bromide replacement. AB - Almost no single commercially available or registered compound is able to replace the broad spectrum activity of methyl bromide. However, combinations of for long known fumigants or their generators, showing good activity in the control of specific groups of soil borne diseases or pests, show interesting broad spectrum activity and even synergism. Well-known combinations are 1,3-dichloropropene (1,3 D) and chloropicrin, applied by injection or through irrigation lines. A few decades ago, combination products containing 1,3-D and methyl isothiocyanate (MITC) were developed and commercialized but were not Longer supported. As 1,3-D is an excellent nematicide with other target groups depending on contact time and tarping, and dazomet as a MITC generator shows fungicidal, nematicidal, insecticidal and herbicidal activities, studies with combinations of both were started again. Laboratory trials with different combinations of 1,3-D and Basamid (97 % dazomet) in soil columns showed interesting results both with Basamid solved in 1,3-D, although limited by temperature, and with simultaneously applied products, even at half dose of each, or even tess. In a further stage, small greenhouse trials were performed with tomato and lettuce crop and could confirm synergistic activity through the evaluation of crop stand and harvest results. Other trial work was performed in collaboration between Certis Europe branches and local research groups in both Italy and Spain. There again it was demonstrated that the combination of 1,3-D (injectable as well as emulsifiable formulations) with Basamid are able to control different phytosanitary problems in different crops. In parallel with the above mentioned trial work, emphasis is also made on careful and appropriate tarping of the soil, preferentially by improved gastight barrier film. The latter allows dose reduction as well as improvement of the fumigation performance. Barrier properties of commercially available fumigation film are now studied for both 1,3-D and MITC. Finally, it is emphasized that appropriate application systems and machinery need more attention to achieve good results with this type of fumigants, i.e. less volatile and with less good diffusion capacity as compared to methyl bromide. PMID- 18399426 TI - The effect of air support on droplet characteristics and spray drift. AB - Air assistance on field sprayers creates a forced airstream under the spray boom which blows the spray droplets into the crop. The advantages of this relative new technique are less drift of spray droplets and the possibility to reduce the amount of pesticides and spray Liquid. The purpose of this work was to investigate the effect of air assistance on the characteristics of spray droplets and their driftability. Based on air velocity measurements on an air assisted field sprayer, a system of air assistance was developed in addition to a laser based measuring set-up for the characterisation of spray droplets. With this set up, the effect of air support on the droplet characteristics was investigated for different settings of the air assistance. The effect on spray drift was quantified based on field drift measurements. A reducing effect on the total amount of spray drift was demonstrated for the Hardi ISO F 110 02, F 110 03 and LD 110 02 nozzles with drift reduction factors a(d) of, respectively, 2.08, 1.77 and 1.53. The use of air support had no significant effect for the LD 110 03 nozzles on the total amount of spray drift. Comparing droplet size and drift results, it was found that air support has the highest impact on the amount of spray drift for the finer sprays by increasing droplet velocities. The effect of air support on droplet sizes is rather limited. PMID- 18399427 TI - Optimizing the application of entomopathogenic nematodes: experimental set-up. AB - The complex issue concerning the spray application of Entomopathogenic Nematodes (EPNs) with a hydraulic sprayer is still not solved. This research project focuses on the effect of spray application technique on the viability and deposition of EPNs. In this paper the experimental set-up used for this evaluation is described. A modular spray application system has been developed and is currently used to evaluate the effect of different parts of a sprayer on the viability of the EPNs. Based on the results of experiments using this modular spray application system, recommendations regarding pump type, mixing system, nozzle type and filter size will be formulated. Because of the large number of experiments in this research project, an image analysis system for the determination of the viability of the nematodes is developed. This paper describes two experiments comparing the new developed image processing technique with the standard microscopic counting technique. PMID- 18399428 TI - Personal protective equipment for registration purposes of pesticides. AB - Regulatory authorities in North America, Europe and Australia use different approaches for the estimation of exposure reduction effectiveness of personal protective equipment (PPE) in registration processes of agrochemical pesticides. TNO has investigated current views and facts for the use of default values and set up a discussion paper which can be used as a starting point to achieve an internationally harmonised set of PPE protection factors for regulatory use. For inhalation exposure Loading it is proposed to use the assigned protection factors (APF) as deduced by BSI (British Standard Institution) and ANSI (American National Standards Institution). Since these values are somewhat variance and since in agricultural settings efficient control and proper training and education with respect to inhalation protection devices is generally absent, it is good to err on the safe side and to use the Lowest of both values, if available. For dermal exposure Loading differentiations are made for operators and re-entry workers and further for hand and body protection. Next to this the restrictions and framework for the use of the proposed defaults are very relevant. Oral exposure loading is only considered in special cases where dermal exposure may be relatively high and the hand-mouth shunt may lead to appreciable oral exposure loading. The presented defaults for PPE have been discussed with experts of regulatory authorities and industry, but a formal discussion still has to take place. This needs to be done on EU level between Member States. The current proposal is based on state-of-the-art knowledge and policy considerations, but further research is needed to better underpin the proposed values and/or to adapt them. PMID- 18399429 TI - A rapid microbiotest for determination of soil toxicity to higher plants. PMID- 18399430 TI - Pesticides selectivity list to beneficial arthropods in four field vegetable crops. AB - Selectivity of pesticides to beneficial arthropods is a key data for the implementation of IPM program. In the context of field vegetables crop, a set of 16 fungicides, 17 herbicides and 14 insecticides commonly used in Belgium were tested on 5 indicator species: the parasitic hymenoptera Aphidius rhopalosiphi (De Stefani-Perez) (Hym., Aphidiidae), the aphid foliage dwelling predators Adalia bipunctata (L.) (Col., Coccinellidae) and Episyrphus balteatus (Dipt., Syrphidae) and the ground-dwelling predators Aleochara bilineata (Col., Staphyllinidae) and Bembidion lampros (Col., Carabidae). Pesticides were tested according a testing scheme including a first assessment on inert substrate (glass plates for adults of A. rhopalosiphi, larvae of A. bipunctata and E. balteatus, sand on adults of A. bilineata and B. lampros) and, for product that were toxic, a second assessment on natural substrate (barley seedlings for A. rhopalosiphi, french bean plants for A. bipunctato and E. balteatus and two type of soil for 8. lampros and A. bilineato). The effects of the product were assessed on basis on mortality, except for A. bilineata (Onion fly pupae parasitism). According to the final results obtained at the end of this testing scheme, the product were listed in toxicity class: green list if effect < or =30%, yellow list 30% < effect < 60% and orange list 60% < effect < or =80%. Products with toxicity higher than 80% on plants or on soils, or that reduce parasitism more than 80% on soil were put in red list and are not recommended for IPM. Results showed that all fungicides and herbicides were included in the green list except tebuconazole and boscalid + pyraclostrobin that were labeled as yellow for A. bipunctata. In opposite, no foliar insecticide was totally selective for all beneficial tested. However some products are in green list for one or several species. Soil insecticides were all are very toxic for ground dwelling arthropods and classed in red list. All results obtained during this study and further upgrade will be available on www.cra.wallonie.be/selectivite. In conclusions, fungicides and herbicides tested are compatible with IPM programs. For foliar insecticides, some treatments can be used carefully according to the selectivity. But for soil insecticide treatments, their toxicity raise the question of their use in IPM programs in vegetables and the need of new compounds or development of alternative pest control programs. PMID- 18399431 TI - Influence of organic matter on bio-availability of carbosulfan and its toxicity on a carabid beetle. AB - Study of factors influencing soil insecticide toxicity are needed to reduce negative impacts of these products on beneficial insects. To date, if high toxicity differences between different type of soils have been reported, there is few specific studies on soil parameters influence on selectivity of soil insecticides to beneficial arthropods. To assess the specific impact of organic matter, the relationship between bio-availability of a soil insecticide, carbosulfan [Sheriff 1 Gr], and its toxicity on a small Carabidae, Bembidion lampros (Herbst.) on a sand enriched with increasing quantities of organic matter was studied. In laboratory, adults of B. lampros were put on different substrate, made of sand or sand with addition of organic matter at 3, 6 and 9% w/w, and treated with carbosulfan applied as granule at the rate of 312.5, 625, 1250 and 6250 microg a.i./m corresponding respectively to 0.5, 1, 2 and 10% of the recommended field rate. Mortalities of B. lampros were assessed after 14 day of exposure. In parallel, the total carbosulfan residue (total extraction) and bioavailable fraction (CaCL2 aqueous extraction) were determined 48h after substrate treatments. According to the mortalities and bio-availability obtained, a dose--response relationship was calculated and compared with a reference relation dose--response obtained on sand, where the bio-availability of the product was considered as 100% of the amount of product applied. Carbosulfan was highly toxic on sand for B. lampros, with 100, 57 and 50% mortality at 10, 2 and 1% of the recommended field rate. When organic matter was added to the sand, the toxicity gradually decreased. This reduction in toxicity was more rapidly observed on sand + organic matter than on pure sand. The mortalities were strongly correlated with the bioavailability, indicating first that the organic matter is fixing an important part of the insecticide and secondly reduce its toxicity to beneficial arthropods. The results suggest that it could be possible, with further research, to predict toxicity of products in the field on different kind of soil according previous laboratory toxicity models and soil analysis. In conclusion, the organic matter influences strongly bio-availability of carbosulfan. This bioavailability was strongly correlated to toxicity to B. lampros. With the improvement of bioavailability determination and method validation, the assessment of pesticide bio-availability in the substrate could help to estimate the pesticide toxicity towards carabidae on different type of soils. PMID- 18399432 TI - Residues effects of isoproturon in mature earthworm (Aporrectodea caliginosa) under laboratory conditions. AB - This study was conducted to investigate the residues of isoproturon and its metabolites, 1-(4-isopropylphenyl)-3-methylurea, 1-(4-isopropylphenyl) urea, and 4-isopropylanilin in soil and mature earthworms under laboratory conditions. Mature earthworms (Aporrectodea caliginosa) were exposed for various durations (7, 15, 30, and 60 days) to soils contaminated with isoproturon concentrations (2, 4, 6, 8, and 10 mg.kg(-1) soil). The decrease in isoproturon concentration in the soil depended on initial concentration it was slower at higher concentrations. The isoproturon and its metabolites accumulated in earthworms it increased during the first 15 days and decreased thereafter. Acute toxicity of isoproturon was determined together with total soluble protein content and glycogen of worms. These parameters were related to isoproturon concentration in soil and earthworms. No lethal effect of isoproturon was observed even at the concentration 1000 mg.kg(-1) soil after 60 days of exposure. A reduction of total soluble protein was observed in all treated worms (maximum 59.54%). This study is suggesting the use of the total soluble protein content and glycogen of earthworms as biomarker of exposure to isoproturon. PMID- 18399433 TI - Powder formulation of Burkholderia cepacia for control of rape seed damping-off caused by Rhizoctonia solani. AB - Talc-based formulation of Burkholderia cepaci strain Bu1 was tested as seed and soil drenchs separately for its ability to control Rhizoctonia soloni the causal agent of rape seed damping-off in greenhouse and field trials. In general, the formulated bacteria was more effective to suppress the disease than the suspension of bacteria cells in carboxymethylcellulose solution (1% w/v), in both greenhouse and field trials. The formulation of strain Bul as soil and seed treatments had the greatest effect on reducing the rape seed damping-off in greenhouse and field trials (66.7, 53.3, 64.4 and 40% respectively). The formulation of strain Bu1 as soil and seed treatments were the most effective treatments to increase the root dry weights in the infected soil in greenhouse. The formulation of strain Bul as soil drench had the greatest effect on enhancement of the fresh weight of roots and stem fresh and dry weights. The formulation of strain Bu1 stored at 4 degrees C exhibited better shelf Life and efficacy in vitro than it's counterpart stored at 25 degrees C. PMID- 18399434 TI - The use of HET-CAM test in detecting the ocular irritation. AB - Agrochemicals must undergo numerous toxicological tests before registration. One of these experiments is the examination of eye irritation potential. To get knowledge about eye irritation, recently only the in vivo Draize-test is accepted, which is one of the most criticized methods because of the injuries inflicted on the test animals. Several in vitro methods have been used to investigate the toxicity of potential eye irritants with a view to replacing in vivo eye irritation testing. In the HET-CAM test chemicals are placed in direct contact with chorioallantoic membrane of the hen's egg. The occurrence of vascular injury or coagulation in response to a compound is the basis for employing this technique as an indication of the likelihood that a chemical would damage mucous membranes (especially the eye) in vivo. In our studies comparative screening was performed with a set of agrochemicals to establish paralell data on in vitro (HET-CAM) and in vivo (Draize) results in case of 6 agrochemicals. The solutions to be tested are added to the membrane and left in contact for 5 minutes and the membrane is examined for vascular damage at set time periods. Irritancy is scored according to the severity and speed at which damage occurs providing an indication of the likely irritant effect of the compound. Our study showed good correlation between results obtained by the HET-CAM test and those of the Draize rabbit eye test most cases. The present form of the HET-CAM test can be proposed as a pre-screen method of eye irritation tests. PMID- 18399435 TI - The impact of flucycloxuron on eggs weight kinetic and hematological parameters of chicken (Gallus domesticus). AB - The struggle against the harmful bugs of culture is intensified, and several products are appeared every year without the knowledge how to control their effects on environment and especially on being life. The introduced chemical products in nature are generally, the synthesis products witch are the pesticides. Our study consist the impact mechanism of a pesticides (FCX) on other biological model than harmful bugs, this biological model is a vertebrate model witch is the domestic chicken eggs (Gaollus domesticus). The toxicity of Flucycloxuron reviewed across the eggs weight kinetic accompanied with embryonic hematological parameters, in ovo and after hatching. The tested concentrations of pesticide are 1, 10 and 20 microg/egg injected at first day of incubation. Eggs treatment by three concentrations of pesticides, disturbs the studied parameters, where we observe that the pesticide inhibit the nutriment transformation, translated by eggs decreased weight kinetic according to the control, also the FCX affect the shell weight and cause the alteration of shell integrity. Hematological parameters show a clear impact of the pesticide at the lowest concentration (1 microg/egg). The obtained results confirm that the chosen biological model is good bio-indicator for eventual pollution and they are not far from pesticides toxicity. PMID- 18399436 TI - Bioavailability of insect growth regulators in citrus and stone fruits. AB - The aim of this study was to offer data about the bioavailability of flufenoxuron, lufenuron, pyriproxyfen and fenoxycarb in common commodities like mandarin, apricot and peach. The in vitro bioavailability of the compounds was studied not only in fresh fruit but also in standards and canned food in order to establish possible differences according to the matrix. The gastric digestion was simulated with porcine pepsin at pH 2, for 2 h in a shaking water bath at 37 degrees C. The intestinal digestion was simulated with porcine pancreatin at pH 7, for 2 h in a shaking water bath at 37 degrees C. The intestinal absorption was simulated with cellulose dialysis tubing filled with a solution of sodium carbonate. No in vitro bioavailability was observed in mandarin, peach and apricot samples spiked at the concentrations generally found in the market for the raw and processed commodities. In standards, the dialysis started at the level of 0.25 mg/kg. This is an approximation to the pesticide digestion and absorption in humans. PMID- 18399437 TI - Dissipation of insect growth regulators in fresh orange and orange juice. AB - It was studied the dissipation rates of fenoxycarb, Lufenuron, flufenoxuron and pyriproxyfen from their application on navelina orange crops to the production of orange juice. Supervised trials were carried out for the phytosanitary treatments under two situations, one according to Good Agricultural Practices (GAP) and the other one with Critical Agricultural Practices (CAP). Samples of both situations were transformed into orange juice according to the current industrial process. The analytical methodology included acetone and dichloromethane/petroleum ether extraction and aminopropyl-based cleanup. Method validation followed SANCO Guidelines. The final objective was the determination of the exposure to the residues in raw and processed orange when good and critical agricultural conditions are used in the field. PMID- 18399438 TI - Removal of pesticides from white wine by the use of fining agents and filtration. AB - The effects of four clarification agents (bentonite, charcoal, PVPP and potassium caseinate) on the removal of residues of three fungicides (famoxadone, fluquinconazole and trifloxystrobin) applied directly to a racked white wine, elaborated from Airen variety grapes from the D.O. Region of Jumilla (Murcia, Spain) are studied. The clarified wines were filtered with 0.45 microm nylon filters to determine the influence of this winemaking process in the disappearance of fungicide residues. Hydro-alcoholic solutions with the three fungicides at concentrations of 1 and 2 ppm were then added through intense stirring to each of the containers. Two hours later, the corresponding clarifying agent was added with intense stirring for some minutes. The containers were then sealed and left to settle for five days. Once the clean wines had been racked, they were filtered through nylon 0.45 microm pore filters. All assays were performed three times. Analytical determination of fluquinconazole and trifloxystrobin was performed by gas chromatography with an electron captor detector (ECD), while that of famoxadone was made using an HPLC-DAD. For the three fungicides, the highest elimination is produced with the clarification by charcoal, reaching Levels of removal of 100% in all cases. For the four clarifying agents, the highest elimination is produced for the fluquinconazole residues. The fungicide which is retained most in the lees is famoxadone, since it has the lowest solubility of the three pesticides studied. The highest percentage of residues in the lees is obtained for the assay with charcoal. The filtration process of the clarified wines using the four agents studied is not effective, since the elimination percentage is generally lower than 10% of the initial residues in the non-clarified wines. PMID- 18399439 TI - Influence of fungicides on grape yeast content and its evolution in the fermentation. AB - The influence of six fungicides (famoxadone, fenhexamid, fluquinconazole, kresoxim-methyl, quinoxyfen, trifloxystrobin) on the yeast content in harvested grapes has been studied along with their effect on it during the wine-making process. Two treatments were carried out with authorized formulates at the manufacturer doses. The first was carried out under good agricultural practices, obeying the security times, and the second one under critical conditions, applied on the day of harvesting. The grapes were harvested two hours after the application. During the wine-making process, samples were taken at 1, 5, 12 and 20 days after the start. The levels observed in the control sample (1.9 log CFU/cm2) were similar to previous studies. The counts in all treated samples were higher than the control sample, without adverse effect of the pesticides on the yeast count even in the grapes treated on the day of harvest. As with the observations on grapes, no effect of the pesticides on the fermentation count was observed. All counts on the treated samples were higher than the control sample. The treatment on the day of harvest did not have any effect on the fermentation count, with superior results being obtained for all the pesticides. PMID- 18399440 TI - The toxicity study of a systemic fungicide: artea 330 EC on the physiology and the respiratory metabolism of the tadpole (Rana saharica). AB - The intensification of the cereal cultures accompanied by the apparition of damaging illnesses for these cultures. These illnesses are Largely imputed to mushrooms micro and macroscopic chatty of important damages at wheat, the barley and of none targeted other animal species. The products used against these illnesses are called: Fungicides. In our work, we are interested in the survey of the effects of a systemic fungicide: the ARTEA 330 EC introduces newly in Algeria, on some physiological and metabolic parameters of a biologic model: the tadpole The exhibition of the populations of tadpoles to the different concentrations of the ARTEA 330 EC provokes a fall very important of the middleweights of the tadpoles treated. This one is reduced of meadows of 80% to the strongest concentration of ARTEA 330 EC. It is some in the same way for the reduction of the middle size of the tadpoles where one records a reduction of 25%. Concerning the percentage of mortality gotten, we observe that the one is here from 100% to the strongest concentration of ARTEA 330 EC. The breathing of the tadpoles treated by fungicide is disrupted strongly and to the strongest concentration, this one is inhibited completely. The gotten results show that the exhibition of the populations of tadpoles to the different concentrations of fungicide disrupts the physiological parameters strongly and inhibit the respiratory metabolism. A phenomenon of detoxication seems to intervene; it is put in evidence by a stimulation of the synthesis of the proteins. PMID- 18399441 TI - Removal of prioritary pesticides contamining r'mel ground water by using organic waste residues. AB - This study evaluated pesticide contamination of R'mel ground water located in northwest Morocco. The study area is densely populated and thriving, with intensive agriculture. Various techniques, including stir bar sorptive extraction (SBSE) and gas chromatography with mass spectroscopy detection (GC-MS), were used for the quantitative determination of 13 pesticides including alachlor, aldrin, atrazine, chlorpyrifos, chlorfenvinphos, dieldrin, alpha-endosulfan, endrin, hexachlorobenzene, beta-HCH, gamma-HCH (lindane), simazine and trifluralin. The survey results showed that contamination by pesticide residues is widespread in the area. With the exception of atrazine, the average concentrations were all below the regulatory limits established by the European Union. The potential of ten natural organic substances to eliminate pesticides included in the European Water Framework Directive was evaluated. The absorbents with the highest removal efficiency were date and olives stones and, to a Lesser degree, Raphanus raphanistrum and Cistus ladaniferus. The adsorption tests gave very satisfying results and pointed to the possible application of these supports as ecoLogical remediation techniques to prevent pesticide pollution of aquatic ecosystems. PMID- 18399442 TI - Distribution of organochlorine pesticides in crop growing in the province of Latina. AB - Aim of this trial was to verify the occurrence and the distribution of organochlorine pesticides (OCPs) in soil, uptake and translocation by different crops growing in glasshouses with different level of pollutants. Residues of OCPs have been detected in soils and crops in the province of Latina, an intensively agricultural area of Lazio Region, in Italy. Dieldrin was quantified particularly in zucchini, cucumber, melon, tomato during a vegetation period. Sampling was carried out at three different times after transplanting. Roots, stem, Leaves and fruits were analysed separately. It would not be advisable to grow cucurbits in soil with OCPs residues > 0.100 mg/kg, to respect the RML admitted in UE in cucurbitaceous fruits. PMID- 18399443 TI - Assessment of side-effect of water-soluble nitrogen fertilisers applied as foliar spray on the parasitic wasp Aphidius rhopalosiphi (Destefani-Perez) (Hym: Aphidiidae). PMID- 18399444 TI - Threats and control of the brown necked ravens (Corvus ruficollis) in Egypt. AB - The Suez Canal region is a small fertile area in Egypt that is placed under enormous pressure of existing resources. In these areas, intensive agricultural practices are performed in combination with high population densities. The described problem is that together with agricultural practices and urbanization, Brown necked ravens (Corvus ruficollis) are in huge amounts attracted. The birds are very aggressive and showed to be a serious pest. They spread germs and diseases for human health and food production. Therefore, the birds need to be controlled. Our research focuses on identifying food preferences of raven and on the way raven control is most effective. Ravens are omnivorous birds. From our laboratory study it was seen that most preferable foods were, in descending order: fresh fishes, cow liver, crustacean, watermelons, tomatoes and yoghurt. Under field conditions where stomach content was dissected, animal matters showed to a more preferred food source than plant matters. Observations on olfactory sensitivity showed that ravens could easily locate their food. Biological observations in the field on reproduction of ravens showed that raven females lay two to six eggs. Average number of babies per nest was between one and four. Average number of raven flock before sun rise was more than 100, while it was less than 100 before sun set. The impact of mechanical, biological and chemical control was investigated. Without control, approximately flock numbers of more than 100 ravens were recorded. The average number of raven flock was 60 before mechanical control operation (nests destroyed and using net), while it was 40 after mechanical control. Results of the biological control showed that kestrel (Falco tinnunculus rupicolaeformes) predated raven babies more effective than barn owls (Tyto alba). Within the chemical control experiments, Brodifacoum (0.0005%) was most effective against ravens, followed by Zink phosphide (19%) and Methomyl (90%, carbamate compound). PMID- 18399445 TI - Survey of weed composition befor maize sowing in long-term fertilization experiment. AB - The study was carried out in a long-term fertilization field experiment of the Experimental Station of University of Pannonia, Department of Crop Science and Soil Science in 2006. The Long-term fertilization experiment was set up in 1983. In the experiment, the success of the weeds ability to grow under the influence of NPK, NPK + FYM* and NPK + straw treatments was compared, and the effect of increasing Nitrogen dosing on weediness was studied. The bifactorial test was arranged in split plot design with three replications. Treatment A: nutrient: NPK, NPK + 35 t/ha FYM* and NPK + straw manure. Treatment B: N kg/ha(-1) N0-N4 (0, 70, 140, 210, 280), and 100 kg P2O5 ha(-1) & 100 kg K20. The weed survey was made on 2nd of May 2006. There were spraying no herbicide until the survey. For the weed survey the Balazs-Ujvarosi coenological method was applied. Altogether, we have found 23 weed species in the trial. In the NPK treatment there were 20 species, in the treatment NPK+organic manure there were 17 species and in the NPK+ stalk rest treatment there were 16 weed species. The most dominant of the weeds on the NPK and NPK+straw manure treatments was Veronica hederifolia while on the fertilizer + FYM, the A. theophrasti was most dominant. The average weed covering value of the treatment NPK + FYM was 1.36 times higher (10.87%) than that of treatment NPK only (7.97%) and 3.65 times higher than on the NPK + straw manure treatment. PMID- 18399446 TI - Chemical control of plant habitus in summer-to-autumn flowering chrysanthemum (Dendranthema grandiflora). AB - Summer-to-autumn flowering chrysanthemums (Dendranthema grandiflora Tzvelev. cv.) are grown outdoors in containers and are characterised by a uniform ball-shaped growth habit. Development of new cultivars expanded the production of marketable plants from end of July until end of October. During the first weeks of the production cycle plants were pinched regular to increase branching but also to prevent development of precocious initiated flowers. This pinching treatment however increases Labour costs and is therefore more and more abandoned. The effectiveness of ethephon for terminal bud destruction was evaluated for three cultivars 'Draga', 'Tardero' and 'Veria Dark'. PLants were sprayed with 0-240-480 1200-2400 mg a.i./l. Ethephon spraying resulted in growth arrest and apical bud necrosis and higher doses resulted in more pronounced reactions. The highest dose (2400 mg a.i./l) resulted in phytotoxicity one week after application; phytotoxicity was more severe in 'Veria Dark' than 'Draga' and 'Tardero'. Ethephon treatments significantly delayed flowering and have potential to be used in commercial production schemes of summer-to-autumn flowering chrysanthemum in outdoor conditions. PMID- 18399447 TI - A new weed control strategy in onion culture. AB - A new strategy combining modem hoeing technique and spray application has been developed in order to reduce the amount of herbicides down to 20% compared to common practice. The effects on weed control have been investigated as well as the impact on qualitative and quantitative harvest. In two large scale field trials and two years of testing the authors evaluated different hoeing techniques combined with band spray application and standard spray application, the minimal lethal herbicide dose method (MLHD). All varieties have been calculated for environmental impact as well as practical and economical means. These studies reveal crop losses due to improper weed control as well as losses due to herbicide stress. Detailed information on concentration depending impact of several herbicides have been correlated to their control of different weeds and the achieved yield. Two contrary effects influencing the total yield have been identified. The novel strategy is based on the knowledge of these complex effects which finally led to a well practicable and highly economic strategy that enables onion farmers to control weeds while reducing the amounts of herbicides down to approximately 20%. PMID- 18399448 TI - The phytotestkit: a practical tool for early growth measurements in plants. PMID- 18399449 TI - Chemical weed management in wheat at higher altitudes. AB - To evaluate the effect of different herbicides for controlling weeds in wheat (variety Fakhr-i-Sarhad),at higher attitude, an experiment was conducted at Agriculture Research Station, Chitral during Rabi season 2003-04, using Randomized Complete Block Design, keeping four replications. The experiment, sown in November comprised of eight treatments, viz; seven herbicides and a weedy check. Each treatment consisted of 5 rows each 30 cm apart and 5 m long thus giving a total size of 5 m x 1.5 m. The herbicides used included; terbutryn + triasulfuron at 0.16 kg, 2,4-D at 0.7 kg, fenoxaprop-P-ethyl at 0.93 kg, clodinafop at 0.05 kg, bromoxynil + MCPA at 0.49 kg, carfentrazon-ethyl at 0.02 kg and isoproturon at 1.0 kg a.i ha(-1). The data were recorded on weed kill efficiency (%), fresh weed biomass (kg ha(-1)), plant height (cm), spike length (cm), number of tillers m(-2), number of grains spike(-1), thousand grains weight (g), biological yield (kg ha(-1)), grain yield (kg ha(-1)) and harvest index (%). The data recorded on weed kill efficiency, weed biomass (kg ha1), grains yield (kg ha(-1)) and harvest index (%) were significantly affected by the different herbicidal treatments. Statistically isoproturon treatment exhibited the best performance, with maximum weed kill efficiency (48.26%) and minimum fresh weed biomass (433.3 kg ha(-1)) as compared to weedy check (6 %) and (1102 kg ha(-1)), respectively. Similarly, the spike length (8.34 cm), number of tillers (427 m( 2)), number of grains spike(-1) (38.0), thousand grains weight (39.85 g), biological yield (8475 kg ha(-1)), grain yield (2530 kg ha(-1)) and harvest index (31.3%) were the highest in isoproturon treatments as compared to weedy check having (7.64 cm), (356 m(-2)), (34.1), (37.12 g), (6858 kg), (1913 kg ha(-1)) and (27%), respectively. PMID- 18399450 TI - Effect of selected sugar beet herbicides on germination of various Chenopodium album populations. AB - Seeds of various fat-hen populations (Chenopodium album L.), mostly originating from sugar beet fields, were subjected to treatments with the following herbicides: metamitron, acetochlor, dimethenamid-P and S-metolachlor. Herbicides were applied either incorporated into a sandy Loam soil (2005-2007) and/or on filter paper in Petri dishes (2006-2007). Results between experiments were highly contrasting. Soil applications of metamitron, acetochlor and S-metolachlor were stimulating germination in the 2005 experiments, whereas in the 2006-2007 experiments effects were ranging from slightly stimulating to highly inhibitory. PMID- 18399451 TI - No-tillage system applied to the sunflower (hybrid pioneer PR64E83) resistant to the tribenuron-methyl in the conditions from Romania. AB - In the year 2006, the first experience with the NO-TILLAGE system for sunflower crops was organized in Romania, using the Pioneer PR64E83 sunflower hybrid resistant to the tribenuron-methyl herbicide. The experience took place in the Danube river flood plain, on an alluvial soil with a content between 3.1-4.5% and a content of clay of 34-40%. In the conventional system, in the fall of the year 2005, autumn ploughing was carried out, along with harrowing, and in spring the land was tilled two more times, using both the disk and the harrow. Before sowing, the land was prepared by using the combiner-machine. After the sunflower sprouted, three-time mechanical hoeing between the rows, as well as three-time manual hoeing on the sunflower row were performed. In the no-tillage system, no autumn ploughing, nor any other soil work were performed, and the sunflower was sowed directly, by using the Gaspardo seeder. Before sowing, the soil was covered in proportion of 70% by species of weeds of the Cirsum, Sonchus and Polygonum amphibium type. To destroy these species of weeds, 3 days before sowing, the soil was treated with the Roundup herbicide in a dose of 7 litres/ha. After the sunflower sprouted, when the plants had 4-5 leaves, the following sulphonylurea herbicides were applied post-emergently. (1) Express 50SG--which contains tribenuron-methyl (2) Granstar 75DF--which contains 75% tribenuron-methyl (3) Titus 25DF--which contains 25% rimsulfuron (4) Mistral--which contains 40 g/L nicosulfuron (5) Lintur 70WG--which contains 4.1% triasulfuron + 65.9% dicamba (6) Peak 75WG--which contains 75% prosulfuron The Express 50SG and Granstar 75DF herbicides were selective for the sunflower hybrid PR64E83, and the other herbicides although belonging o the same sulphonylurea group, they proved to have a very phytotoxic effect upon the sunflower crop. In the report, selectivity is presented depending on the dose of herbicide, weed control and sunflower production. PMID- 18399452 TI - The analysis of the effect of different herbicides on Johnson-grass in maize. AB - The Johnson-grass (Sorghum halepense /L./Pers) is a perennial monocotyledon, its gen centre is in the Near East. Botanically this weed belongs to the monocotyledone class, Poaceae (Gromineae) family and within this, together with the maize to the Andropogonoideaei subfamily. At the time of the First National Weed Survey (1947-1953) it has not been found in Hungary. In the periods of the 2nd (1969-1971), 3rd (1987-1988) and 4th Survey it occupied the 94th, 18th and 10th place of importance, respectively. Because of its rapid multiplication and spreading after the second weed survey (1969-1971) and because of its very complicated and difficult control, the Plant Protection Division of our Ministry of Agriculture ranged it in 1974 into the category of "Dangerous weed". Johnson grass is the 6th most important weed in the world. A small plot field trial was carried out in Szabadszentkirtly (12th May 2006) in maize. The effect of pre- and post emergent herbicides was investigated against Johnson grass germinating from seed and sprouting from rhizome. The treatments were done in 4 repeats with Stomp 330 EC, Dual Gold 960 EC, Merlin WG, Wing EC, Guardian Max and Monsoon herbicides. The number of weeds germinating after spraying and the number of damaged weeds were counted and compared to the control plots 10 days after the treatments and then weekly. Parallel to the field trial, a "soil culture in pots" trial in glasshouse was also carried out in which the effect of the above mentioned herbicides was investigated against Johnson grass germinating from seed. During on experiments all the herbicides showed an excellent herbicide activity until the 2nd week of treatment, although the precipitation of the second week was only 8.4 mm. Under the given experimental conditions the best herbicide effect until the 4th week, in both experimental phases were given by Merlin WG. PMID- 18399453 TI - Geographical distribution of principal cereal weeds in setif high plains. AB - The invasion of the wheat and barley cultures by weeds remains among the major problems of agriculture in Algeria considering the importance of yield tosses caused by these species. The cereal yields average is less than 700 kg ha(-1). The main purpose of this work was to obtain informations on geographical distribution of principal cereal weeds in the setif high plains. Two bioclimatic areas may be distinguished in this region, the semi-arid in the south and the sub humid in the north. One hundred ten fields of cereals were surveyed by stratified sampling. In each field the level of infestation was scored on a homogeneous surface. The weediness of the species was estimated on the basis of the frequency and average of abundance. The results indicate thirty four important species. This harmful flora was characterized by the prevalence of annual and brod leaved weeds. The study of the geographical distribution cards of these species shows three groups: species related to the northern zone, species related to the southern zone and species common to both zone. The knowledge of the geographical distribution of principal weeds is an essential element for any plan of integrated weed management. PMID- 18399454 TI - Comperative investigations of non chemical weed management methods in Hungary. AB - Organic farming has an increasing tendency in Hungary because of growing consumers' demands according to organic products not only in inland but also in the countries of the European Union. Developments of weed control methods in organically cropped field plants have become conspicuous next to developing chemical weed management methods of convencionally cropped cultural plants. The aim of our investigations was to make comperative investigations of non chemical weed control methods in wide rowed plants. PMID- 18399455 TI - Characterisation of powdery mildew resistance in a segregating diploid rose population. AB - Powdery mildew (Podosphaera pannoso) is one of the most serious fungal diseases on both greenhouse and field grown roses. Improvement of disease resistance is a major selection aim for garden rose breeders. For rose cultivars, being mostly tetraptoid, it is complicated to develop molecular markers for resistance. Hence, a segregating diploid population was established from a cross between 'Yesterday', a commercial available rose variety susceptible to powdery mildew, and R. wichurana, a rose species with resistance to certain isolates of powdery mildew. A progeny of 94 seedlings was planted in the field. The segregation of powdery mildew resistance was studied in this population by means of a bioassay with two different monoconidial isolates of powdery mildew. Based on the response to these inoculations different groups were selected: a first group of genotypes was susceptible to both isolates, other groups were susceptible to one of both isolates and a last group was resistant to both tested isolates. The disease resistance inherits for both isolates in a quantitative way. A genetic map based on AFLP and SSR markers was established and will be used for QTL analysis of powdery mildew resistance. PMID- 18399456 TI - Accumulation of pearl millet downy mildew resistance in Mali--2006 results. AB - Few crop breeding programs today are breeding crops in their areas of diversity and origin. This study reports on a Malian breeding program in an area of genetic diversity. It has the objective to accumulate resistance to major populations of Sclerospora graminicola (= Sg) with modern breeding and selection methods. This study is part of the development of pearl millet top cross hybrids, with a reduced plant height, Sg-resistance (= resistance to pearl millet downy mildew) and 'stay green' at physiological maturity. The parent entries, among other relevant characteristics, were selected for a high level of resistance (good sources of resistance) making use of a combination of artificial young plant screening methods and single location field testing, in 1998. Pedigree selection in F1 to F4 was from 1999 to 2002. Its synthetics and composites were selected for low S. graminicola-levels, in 2003 to 2005 and in 2003 and 2006 tested for S. graminicola-resistance together with 5 checks at two Locations differing in S. graminicola-virulence responses. The 2006 test seemingly indicated the expected quadratic checks, whereby entry 1 is resistant at location 1 and susceptible at location 2 and entry 2 is susceptible at location 1 but resistant at location 2. This quadratic check is indicating differences in virulence between the two S. grominicola-populations and also an adaptation of the pathogen populations on the newly accumulated genes for resistance in the host. It is also indicating that one or more genes for resistance against each of the two populations were accumulated. A good number of synthetics and composites combined low S. graminicola-incidences with relatively high yields and some had 'stay green' at physiological maturity. One too late entry seemingly had immunity. The 2006 results indicate presence of several S. graminicola-resistance genes in the parent entries and accumulation of one or more genes in certain derived entries, and were obtained in combination with reduced plant height and for the first time in pearl millet also with 'stay green' at physiological maturity. The accumulation of S. graminicola-resistance is expected to increase the chance for regional or global 'stay green' hybrids for grain (medium tall) and fodder (tall). PMID- 18399457 TI - Riboflavin-induced resistance against rice sheath blight functions through the potentiation of lignin formation and jasmonic acid signalling pathway. PMID- 18399458 TI - Rationalization of pesticide treatments against powdery mildew of grape. AB - The powdery mildew represents one of the diseases which affect the grape, it is diffused in all agricultural regions with variable intensity and epidemic course in operation of many microclimatic factors. The powdery mildew of grape is caused from Uncinala necator (Schw.) Burr. (nowadays named Erysiphe necator Schwein.); it is controlled with systemic therapy and contact chemicaL products. In some vineyards located in Latium (central Italy), different field trials have been carried out purposely to rationalize the treatments against E. necator. We have studied the powdery mildew infections through monitoring a set of environmental parameters, the evaluation of cultivar sensibility, the agricultural production method and the area characteristics. We have analysed the following environmental parameters monitoring every 15 minutes: precipitation, soil temperature, solar radiation, wind direction, wind speed, atmospheric relative humidity, atmospheric temperature, leaf wetness, soil humidity to cm 20 and soil humidity to cm 40. Besides, we have used Artificial Intelligence analysis techniques to try to forecast U. necator infections. Guideline EPPO/OEPP PP 1/4 (4) has been used. The trials were conducted in conventional and organic farms. In 2 conventional farms and in organic farm we have considered 1 untreated control thesis, in order to follow the course of infection, 1 standard farm reference thesis (standard), where the treatments were carried out according to the usual farm procedures and 1 thesis where the treatments were carried out according to examining the environmental data. In another conventional vineyard, we have considered only 1 untreated control thesis and 1 standard farm reference thesis (standard) to study disease trend. The achieved results have underlined the possibility (through the knowledge of data pedoclimatic and cultural) to position the treatments against the powdery mildew so that to reduce their number. The lower number of treatments that could follow as a result of environmental data to analyse could bring a series of evident economic and ecologic advantages for the farms. PMID- 18399459 TI - A two years study results in the use of artificial neural networks to forecast Plasmopara viticola infection in viticulture. AB - This paper describes the further results of the study that has been described in session 5 of the 58th International Symposium on Crop Protection (Ghent 2006). Since then our attention has been focused on verifying the previous communication results working on a two years basis data set belonging to a specific farm. The choice of using data from a single farm derives from the considerations that have been explained in the previous study in which it was clear that an efficient forecasting Artificial Neural Network (ANN) model can be created only in restricted (or at least comparable) pedoclimatic areas. On the basis of the matured experience, at the moment we have realized an ANN which, being trained on 2005 year data, elaborating the following year data is capable of correctly predicting the real Plasmopara viticola (Berk. et Curt.) Berl. et De Toni outbreak, never giving false negative signals (no alarm in presence of infection on the field) and, finally, giving few other alarms which are totally comparable with the ones given by the most common statistical instrument used in this field trials. We confirm the advantages of this approach in terms of: (a) Management and optimization improvement of agricultural activities. (b) Reduction of plant protection products use. (c) Quality improvement of the final product for a real lowering of plant protection products use. (d) Reduction of environmental impact. (e) A more efficient management of the climate changes. PMID- 18399461 TI - Barley yellow mosaic virus is overcoming RYM4 resistance in Belgium. AB - Barley yellow mosaic virus (BaYMV) is the causal agent of a soil-borne systemic mosaic disease on barley. It has been reported in Belgium since the 1980s. The control of this disease is managed almost exclusively through the use of resistant varieties. The resistance of most commercial barley cultivars grown in Europe is conferred mainly by a single recessive gene, rym4. This monogenic resistance provides immunity against BaYMV pathotype 1 and has been mapped on barley chromosome 3HL and shown to be caused by mutations in the translation initiation factor eIF4E. Another pathotype, BaYMV pathotype 2, which appeared in the late 1980s (in Belgium, in the early 1990s), is able to overcome the rym4 controlled resistance. Until recently, this pathotype remained confined to specific locations. During a systematic survey in 2003, mosaic symptoms were observed only on susceptible barley cultivars collected in Belgian fields. BaYMV was detected by ELISA and RT-PCR on the susceptible cultivars and only by RT-PCR on the resistant cultivars. In 2004, mosaic symptoms were observed on susceptible and resistant cultivars. BaYMV was detected by ELISA and RT-PCR on both cultivars. In addition to developing RT-PCR methods for detecting and identifying BaYMV and Barley mild mosaic virus (BaMMV), an RT-PCR targeting the VPg/NIa viral protein part of the genome, known to discriminate the two BaYMV pathotypes, was set up to accurately identify the pathotype(s) now present in Belgium. The sequences from the generated amplicons revealed the single nucleotide substitution resulting in an amino acid change from lysine to asparagine specific to BaYMV pathotype 2. The possible reasons for the change in the BaYMV pathotype situation in Belgium, such as climatic change or a progressive build-up of soil inoculum potential, will be discussed, as well as the use of eIF4E-based resistance. PMID- 18399460 TI - Low levels of ochratocin A in wines from Piedmont. AB - Ochratoxin A (OTA) is a mycotoxin mainly produced by a number of species of Aspergillus, commonly found in warm and tropical climates. OTA poses risks for the human health because of its nephrotoxic, teratogenic, immunotoxic and neurotoxic activity. The mycotoxin, classified as possible human carcinogen (Group 2B) by the IARC, naturally occurs in a wide range of foods, including wine, where the main producer is A. carbonarius. The aim of this work was the validation of a procedure for the analysis of OTA in Piedmontese red and white wines produced after vintage 2003 and 2004, in relationship with the limit of 2.0 microg l(-1) introduced by European Union for wine, must or grape juice (Regulation CE N. 123/2005). An analytical method based on immunoaffinity column (IAC) for clean-up and liquid chromatography with fluorescence detection (LC-FLD) was used to determine the occurrence of OTA in wines. Detection limit (LOD) and quantification Limit (LOQ) were 7.18 pg/ml and 9.31 pg/ml based on statistical method (IUPAC). Average recoveries of OTA from wine samples spiked at levels from 0.1 to 10 ng/ml ranged from 90.8% to 92.4%, with relative standard deviations (RSDs) between 2.64 and 2.71%. Repeatability limit was 8.73 pg/ml for samples spiked with 0.1 ng/ml of OTA. Ninety-one Denomination of Controlled Origin (DOC) wines were analysed, including 41 Barbera (red), 38 Dolcetto (red), and 16 white wines, such as Erbaluce, Cortese and Roero Arneis. The study focused on wines commercialized in Italian supermarkets and wine shops. The white wines resulted, as expected, less contaminated than the red ones. Wines produced after vintage 2003, a season particularly conducive to the growth of A. carbonorius, contained higher levels of OTA than the wines produced in 2004. The samples, resulting positive, contained a concentration of OTA highly inferior to the threshold limits introduced by the European Union. The sample of the highest level of OTA was a Dolcetto produced in 2004, with 1.10 ng/ml of mycotoxin. PMID- 18399462 TI - Effect of composting and pasteurisation on two important quarantine pests of potato. AB - The effect of composting and pasteurization on the quarantine pests of potato Clavibacter michiganensis ssp. sepedonicus (Cms) and Synchytrium endobioticum (Se) were examined on an experimental scale. Composting was performed with 2-L pots and 60-L composters for two months at temperatures below 50 degrees C and for 12 and 21 days at temperatures above 65 degrees C. Pasteurization was performed via water bath at 70 degrees C for maximum 2 hours. Pathogens were introduced directly or via carriers into the processes. After composting for two months and for 12 and 21 days it was possible to isolate vital Cms cells from bioassay plants and vital resting spores of Se could be extracted from sample material. Likewise it was possible to isolate vital Cms cells and resting spores of Se after pasteurization for up to two hours. Both pests could not be killed completely during the performed processes. Further studies concerning sanitization of potato wastes are necessary. PMID- 18399463 TI - Evaluation of innovative products to reduce copper applications to control potato late blight in organic production systems. AB - The main objective of this project, VETAB project, is to determine alternatives to massive copper utilization to control potato late blight (Phytophtora infestans (Mont.) de Bary) in organic systems. To reach such a target, we first performed a screening of candidate products and additives based on their efficiency in the laboratory, under controlled conditions. We evaluated a wide range of products: formulations with a low level of copper, antagonists suspensions, aminoacid extracts, plants extracts, potassium salts, sulphur formulation, organically stabilised peroxide and rhamnolipids. The product's suspensions were applied by vaporization on potato plants. Two different protocols of application were elaborated. To test the fungicide protection action, the product was applied four days before inoculation of the pathogen. To evaluate the defence stimulating effect, the product was applied several times during the plant growth before inoculation of the pathogen. The last vaporization was performed 4 days before inoculation. We also evaluated the resistance of the product to washing risk. Pathogen suspension was applied as droplets of 5 x 10(4) spo/ml on detached leaves. The leaves were then incubated (18 degrees C, RH > 90%, 6 days) in order to record symptoms development. The best results were obtained with formulations integrating reduced doses of copper and with potassium salts. In conclusion, a wide range of products and additives are proposed on the market but very few of those have a real efficiency. The performance of the most efficient products has to be confirmed in field trials. PMID- 18399464 TI - The Cypress aphids in the South of France. AB - In the South of France, Cinara (Cupressobium) cupressi, the main Cypress aphid, has been studied during 6 years on a mixed hedgerow in which Cupressus sempervirens and C. arizonica had been planted alternatively. In the area, this monoecious aphid is anholcyclic and heavily attended by ants. Continuous observation of the trees and of the colonies allowed a description of the population dynamics and the characterization of the damages. The two cypress species are regularly attacked, but the aphid appears earlier, remains longer and is more abundant on C. sempervirens, than on C. arizonica. The distribution of the colonies among trees is contagious. Some trees or groups of trees are attacked more frequently and/or present colonies earlier, suggesting that the tight interaction with ants may induce hivernation and early attack in some places. PMID- 18399465 TI - Using stable isotopes to trace origin and host plants of an African polyphagous pest and an European beneficial insect. AB - Crop protection against insect pests requires first a good knowledge of the biology and ecology of the different pest species and the associated beneficials, in particular the spatial distribution of the populations. But the movement of insect populations in the landscape remains often poorly known and in some cases does not make it possible to know the role of the various cultivated and wild habitats in the dynamics of pest and useful insects. Stable isotopes are a tool contributing to the knowledge of host plants (13carbon/12carbon) as well as geographical origin of insects (1hydrogen/2hydrogen). The analysis of stable isotopic ratios has been performed in south-western France on populations of the hoverfly Episyrphus balteatus, one of the most important predators of the cereal aphids in Europe and on West African populations of the bollworm Helicoverpa armigera, an important polyphagous pest attacking cotton and vegetables in the Old World. Methodology, preliminary results and perspectives given by stable isotopes are presented here. PMID- 18399466 TI - Intraguild interactions of aphidophagous predators in fields: effect of Coccinella septempunctata and Episyrphus balteatus occurrence on aphid infested plants. AB - Intraguild relations between beneficial insects have become a major research topic in biological pest control. In order to understand the intraguild competitions between aphidophagous populations in natural conditions, a field experiment was carried out in the experimental farm of the Gembloux Agricultural University. As biological control of pests involve a community of diverse natural enemies, this experiment firstly aimed to assess the aphidophagous predator diversity and abundance in green pea (Pisum sativum) field and secondly to investigate the impact of the large natural occurrence of C. septempunctata on the aphidophagous beneficial dispersion and efficiency as aphid biological control agents in pea field. Visual observations were weekly performed throughout the 2006 growing season. The pea aphids were attacked by several predatory groups, mainly ladybird beetles and hoverflies. Higher densities of ladybirds and hoverflies were recorded in the beginning of July, associated with an aphid occurrence peak. Using net cage system in the field, the particular intraguild relations between added C. septempunctata or E. balteatus and the natural beneficial arrivals and dispersion were observed. The E. batteatus (eggs and larvae) presence inhibited other aphidophagous predators presence on the aphid infested plants. Lower abundance of E. balteatus was observed on aphid infested plants already colonised by C. septempunctata. To explore more accurately the oviposition and predation behaviours of ladybirds and hoverflies and to determine the chemical factors that could influence these behaviours, current researches are performed in laboratory and will be discussed to promote efficient biological control of aphids by natural enemies. PMID- 18399467 TI - Foraging ecology of aphidophagous predators against Aphis punicae Passerini. AB - Foraging behaviour of three aphidophagous predators, viz., Cheilomenes sexmaculata (Fabricius), Scymnus sp. (Coleoptera: Coccinellidae) and Paragus serratus (Fabricius) (Diptera: Syrphidae) on prey Aphis punicae Passerini (Homoptera: Aphididae) was studied in the laboratory at Indian Institute of Horticultural Research, Bangalore (12 degrees 58' N; 77 degrees 35'E) during 2002 03 to understand the predation dynamics. The foraging studies of grubs of C. sexmaculata and Scymnus sp. on A. punicae revealed that the distance travelled to reach the food (aphids) was negatively correlated with the feeding time of prey (r = -0.434 and -0.743, respectively). The time taken for second encounter was positively correlated with feeding time in case of both C. sexmaculata (r = 0.715) and P.serratus (r = 0.641). The time taken for first encounter of prey by Scymnus sp. showed negative correlation with the feeding time (r = -0.9491) and positive correlation with the distance traveled by the predator to reach the food (r = 0.620). The foraging behavioural studies of syrphid, P. serratus on A. punicae showed that the time taken for first encounter of prey was positively correlated with both the feeding time (r = 0.467) and the distance travelled by the syrphid to reach the food (r = 0.485). The time taken for first encounter of prey also showed significant positive correlation with the search time of the prey (r = 0.384). PMID- 18399468 TI - Pathogenicity of entomopathogenic fungi on hibernating pupae of Cameraria ohridella Deschka & Dimic 1986 (Lepidoptera, Gracillariidae). Part 1: Pathogenicity against the naked pupa. AB - Strains from Paecilomyces fumosoroseus, Lecanicillium muscarium, Metarhizium anisopliae and Beauveria bassiana were examined in standardized Biotest to control the horse-chestnut leaf miner (Cameraria ohridella) in her pupal stage in winter. The fungi were pathogenic against the hibernating pupae of Cameraria ohridella at dose of 1.9 x 10(7) conidia/ml. They were aggressive, led to infection, death and mouldiness of naked pupae. Even at low temperature of 5 degrees C and 12 degrees C. L. muscarium strain V24 showed the highest pathogenicity after 4 weeks against this host, close followed by P. fumosoroseus strain P6. M. anisopliae strain 72 and 8. bassiana strain B412 were also pathogen but after a long-time period. Experiments gave information for general susceptibility of naked pupae of C. ohridella under low temperatures against entomopathogenic fungi. In further examinations it has to be tested, whether fungi can infected, when the pupae stay in their natural surroundings, the pupal cell in the leaf. PMID- 18399469 TI - Pathogenicity of entomopathogenic fungi on hibernating pupae of Cameraria ohridella Deschka & Dimic 1986 (Lepidoptera, Gracillariidae). Part 2: Efficacy of entomopathogenic fungi against pupa in her pupal cell. AB - The naked pupae of chestnut leafminer Cameraria ohridella are susceptible for entomopathogenic fungi Paecilomyces fumosoroseus strain P6; Lecanicillium muscarium strain V24; Metarhizium anisopliae strain M72 and Beauveria bassiana strain B412. Their activity at low temperature is advantageous especially for the use in the winter season. (Richter et al., 2007a). Therefore the possibility of infection should be examined in biotest for pupae in their pupal cells on leaf of chestnut at conditions Like in winter. Efficacy of entomopathogenic fungi is decreased, if pupae are in their pupal cells. But fungi, especially L. muscarium, proved the ability to infect pupae of C. ohridella in their pupal cells. Infected pupae moulded in their pupal cells after application. Infection doesn't come from spores. In semi field trials with L. muscarium 60% of pupae were infected in their pupal cells and died after incubation with mouldiness. PMID- 18399470 TI - Evaluation of different insecticides and strategies against Cacopsylla melanoneura (Forster) (Homoptera: Psyllidae), a vector of apple proliferation phytoplasma. PMID- 18399471 TI - Effects of alpha-mangostin from mangosteen pericarp extract and imidacloprid on Nilaparvata lugens (Stal.) and non-target organisms: toxicity and detoxification mechanism. AB - The brown planthopper, Nilaparvato lugens Stat. (BPH) is the most devastating insect pest in rice fields. Outbreaks of BPH, which are resistant to many synthetic insecticides, can cause total rice crop loss. This research was done to evaluate the efficiency of extracts of mangosteen pericarp (Garcina mangostana L.) as an alternative control of BPH Thailand strain. Topical spraying was applied to various stages of nymphal and adult BPH to determine toxicity. An ethanol extract of mangosteen pericarp extract gave the best control of BPH, with LC50 of 4.5% w/v (r2 = 0.95) with 3rd instar BPH nymphs when compared with the other solvents, hexane, acetone and dichloromethane. The active compound, alpha mangostin showed an LC50 of 5.44%w/v (r2 = 0.88). The toxicity of this extract was less than that of Imidacloprid which showed an LC50 of 0.0042% w/v (r2 = 0.99). The toxicity to non-target organisms was determined. This extract showed toxicity to guppies ((LC50 = 2.53 and 4.27 ppm for females and males, respectively; r2 = 0.97 and 0.97, respectively), bees (LC50 = 4.38% w/v, r2 = 0.95) and mice (no oral acute toxicity and no dermal inflammation but showed eye irritation in 1 day which became normal within 3 days). In vitro detoxification enzyme activities of carboxylesterase, acetylcholinesterase and glutathione-s transferase from BPH after 24 hours exposure were also observed. Carboxylesterase showed stronger activity than other enzymes. Toxicity in terms of LC50 values of both the extract and imidacloprid treatments increased in each generation. The LC50 values for each generation were 4.22-6.67 after sequential spraying. After the ethanol extract was kept at 4 degrees C, room temperature and 55 degrees C for 3 months, the quantity of alpha-mangostin and the BPH control efficiency was lower at 55 degrees C than those for other temperatures. The results from this research indicate that mangosteen pericarp extract can be an alternative insecticide for the control of BPH, which may possess high efficiency, cause with fewer environmental problems and result in less resistance in the BPH. PMID- 18399472 TI - Management of the pest resistance to Bt plants synthesizing two Cry toxins. PMID- 18399473 TI - Joint action of quercetin with four insecticides on the cotton leaf-worm larvae, Spodoptera littoralis Boisd. (Lep. : Noctuidae) in Egypt. AB - The naturally occurring phytoncidal chemical component of some plant- species are responsible for controlling and/or repelling insects from host plants. The use of natural products, readily available in the environment, to control the cotton leafworm, could be of help in reducing the need for applying the synthetic conventional insecticides . Moreover, crude vegetable oils application is quite safer to the non targets and the environment. The use of sex attractant pheromones as predicative tools to forecast pest population and their potential damage levels in specific crops, has significantly been limited because sex attractants are usually directed to one sex only. Thus, identification of plant constituents that attract S. littoralis Boisd. adults, especially females, and that directly affect their feeding or reproductive behaviour, would greatly expand opportunities for manipulation of the biological and environmental events that prevent the establishment, development, and dispersal of the insect-pest under study. Extracted Volatiles from certain parts of cotton plants, attracted both sexes of the cotton leafworm moths instead of attracting one sex only as the pheromones do. Plant flavonoids have been shown by many investigators to have an effect on insect behaviour, growth, and development. Quercetin is one of many bioflavonoids that exists in several fruits and vegetables. The Aim of the present work is to study in vivo the biochemical mode of action of quercetin as a synergist in combination with insecticides. The present results proved the synergistic effect of quercetin when combined with four insecticides, namely, profenofos (organophosphates), deltamethrin (pyrethroid), and tebufenozide et hexaflumuron as insect growth regulators against the studied insect-pest. Quercetin role as a synergist might be attributed to its ability to inhibit certain active oxidases, which may be responsible for in vivo detoxification of the intact insecticides when applied to insect larvae, through glutathione-S transferase. PMID- 18399474 TI - Acarological diagnostic research at the Diagnostic Centre for Plants during the period 2004-2006. AB - During the period 2004-2006, 1691 samples of different origin were examined at the Diagnostic Centre for Plants. We received 1046 samples of imported plant material for detection and identification of quarantine organisms. More than 200 samples were checked on mites and insects to get a phytosanitary certificate for export and 391 samples were investigated for diagnostic reason. The Berlese funnel and dissecting microscopy technique were used to separate mites from the samples. For identification, the mites were slide mounted in Berlese-Hoyer's medium and examined by using phase-contrast microscopy. In 3% of the samples examined on the presence of quarantine organisms, phytophagous mites belonging to the superfamily Tetranychoidea were found, but none with the quarantine status in accordance with the EPPO A1/A2 list. Besides Tetranychus urticae detected on different crops, the cassava green mite Mononychellus progresivus was found on cassava (import Cameroon) in 2006. Tenuipalpus elegans (Tenuipalpidae) was found on cut foliage (import South Africa) in 2004. In 19.9% of the investigated samples for diagnostic reason mites were found. In 47.7% of the infested samples mites were definitely the reason for the damage; in 15.9% mites were secondary and in 36.4% the occurrence of mites was not relevant for the injury. An overview of the determined mites will be given. During this 3 years diagnostic research a few new pest mites belonging to families Tetranychidae and Eriophyidae can be reported. In 2006 Panonychus citri was found on Prunus laurocerasus and later on Eleaegnus sp. and Skimmia sp.. Aceria silvicola was determined on Rubus idaeus in 2006 and Aculus ulae and Aceria carpini on Carpinus betulus in 2005. Besides new pest mites, never seen problems with the broad mite Polyphagotarsonemus latus (Fam. Tarsonemidae) occurred in tree-nurseries in 2005 and 2006. Also 20 samples coming from private persons were investigated. The main problems indoor were caused by Dermanyssus gallinae and Bryobia praetiosa. In gardens especially spider mite problems occurred. Eurytetranychus buxi, Oligonychus ununguis, Eotetranychus carpini and Eotetranychus tiliarium caused considerable damage in topiary and formal fences. PMID- 18399475 TI - Flight patterns of Anarsia lineatella (Lep., Gelechiidae) in relation to degree- days heat accumulation in Northern Greece. AB - The flight patterns of the peach twig borer Anarsia lineatella (Zeller) (Lepidoptera: Gelechiidae) were studied in two important and representative regions of peach production in Northern Greece. In order to determine the seasonal occurrence of the species, adult male moth flight was monitored from April till October (2005-2006) using 8 pheromone traps. The observation areas were located at the regions of Veria (lat. 40.32 degrees North) and Velvendos (lat. 40.16 degrees North). The first flight was observed from early May to early June in Veria and from late May to mid June in Velvendos. The 2nd and 3rd flights were observed from late July to early August and from late August to late September in both regions. The degree-days required for the first generation (431 Veria, 371 Velvendos) were fewer than for the 2nd (661 Veria, 504 Velvendos) and the 3rd generation (675 Veria, 506 Velvendos). A lower threshold temperature of 100 C was used in calculating daily day-degrees from March 1st. All the above data are valuable for the development of prediction models and for decision making in order to achieve better timing of treatments. Considering the increasing interest of biorational insecticides where precise timing of treatments is extremely important, degree-day models could be a useful tool for improving their efficacy in IPM. PMID- 18399476 TI - Strategy of freeze tolerance in adult of sunn pest bug (Eurygaster integriceps Put.). AB - To investigate cold hardiness of Eurygaster integriceps Put, super cooling points of whole bodies during December to March 2006-2007 were measured on samples collected from altitude of Ghara-aghaj varamin-Iran. In addition, the lower lethal temperature (LLT) was determined for adult insect. The super cooling points were observed about -5 in cold months. Since the minimum temperature under natural condition is reached to-7 degreesC or lower in January and LLT99 (-21.65 degrees C), --11 degreesC lower than their minimum SCP (-10.5 degrees C), therefore I concluded that adult insects are freeze tolerant. Furthermore, super cooling points of different sex, weight and attitude were not significantly different. Freeze tolerance in these insects may be a strategy to provide protection from long-term exposures to ice crystals in microhabitat in alititude. PMID- 18399477 TI - Demographic parameters of wrinkling aphid of pistachio leaf, Forda hirsuta Mordv. (Hem.:Pemphigidae) insitu of Rafsanjan's orchards. AB - Wrinkling aphid of pistachio leaf, Forda hirsuta Mordv. (Hem.:Pemphigidae) is one of the pests of pistachio trees. This aphid caused the shrinkage, thickening, and changing the color of the pistachio leaves. Since insect feeds from leave edges, the thick and rolled upward wrinkles were formed, which its green color turned into red. Therefore, its economical damages are out of direct feeding from plant extraction, twisting pistachio leaves, and the decrease of photosynthesis. In this research two orchards and 10 trees that each of them were selected in Rafsanjan region and 58 fundatrix galls, 120 nymphal galls were marked and the demographic parameters for apterus parthenogenesis female of this aphid were calculated via daily observations. The results indicated that intrinsic rates of increase (r) for 1, 2 and 3 generations were 0.01, 0.0638 and 0.0575 femal/femal/days respectively, Doubling time (DT) were 69.31, 10.52 and 12.04 days, respectively, net fecundity rates were 1.71, 11.5 and 7.37 femal/femal/days, respectively, Net fertility rates were 1.11, 8.87 and 5.01 days, respectively, and mean generation times (Tc) were calculated to be 31.5, 32.2 and 31.87 days, respectively. Other reproductive parameters such as gross hatch rate, gross fecundity rate, gross fertility rate, mean age gross fecundity and fertility, mean age net fecundity and fertility, mean age hatch, finite rate of increase (lambda), intrinsic birth rate (b) intrinsic death rate (d) and daily reproductive rate were also calculated. The results revealed that population parameters especially r in the first and second generations were lowest and highest, respectively and mean longevity of fundatrix was 30 days and the born nymphs made separate galls in the edge of pistachio leaves. PMID- 18399478 TI - The occurrence and species composition of thrips (Thysanoptera) on onion. AB - The aim of the present research work was to investigate the population density and species composition of thrips infesting crops of onion in South Poland. The flight activity of thrips was monitored using blue sticky traps and plant samples were taken to record the number of adult and Larvae of thrips on onion. In 2004 the thrips were caught into blue sticky traps from the start of June to the end of the first decade of September. The peak flight activity was noticed in the middle of July. In 2004 the significant growth of the numerousness of adult thrips on onion grown from seeds was recorded in the middle and in the third decade of August, whereas on onion grown from sets in the first decade of August. Thrips larvae were not observed during the whole vegetation season. In 2005, the thrips were caught into blue sticky traps form the half of June to the first decade of September. The peak flight activity was noticed in the first decade of August. In 2005 the rapid growth of the numerousness of adult thrips on onion grown from seeds was recorded in the third decade of July. The highest number of adult thrips on onion grown from sets was noticed at the beginning of August. Thrips larvae were observed on onion grown from seeds in the third decade of July and at the end of the second decade of August. In both years of observations the most numerous species was Frankliniella intonsa Tryb. The second most numerous species was Thrips tabaci Lindeman. In 2004, the most numerous species was predacious Aeolothrips intermedius Bagnall. PMID- 18399479 TI - Analysis of the thrips fauna (Thysanoptera) on flowers of herbs. AB - Studies of Thysanoptera were realized at the Agricultural Experimental Station in Mydlniki near Cracow over the years 2003-2005. This research work covers the observations of the species composition of thrips and the periods of the occurrence of the particulars species of thrips and their numerousness. The thrips were collected from flowers of Valeriana officinalis L., Hypericum perforatum L. and Levisticum officinale Koch. Following thrips species dominated samples collected directly from plants: Frankliniella intonsa (Trybom), Thrips fuscipennis Haliday, Thrips tabaci Lindeman and Thrips major Uzel. The greatest number of thrips was noticed during flowering period on Valeriana officinalis L., and Hypericum perforatum L. PMID- 18399480 TI - How does the age of hoverfly females affect their reproduction? AB - In the present study, we observed the effect of age of Episyrphus balteatus DeGeer (Diptera, Syrphidae) females on their fecundity and fertility (number and percentage of fertile eggs). Eight newborn E. batteatus couples were placed in separated cages (30 cm x 30 cm x 60 cm) and the number of eggs laid and the egg viability were recorded daily during 45 days. E. balteatus females had a pre oviposition period of 10 days before mating happened. The fecundity increased steadily from day 11 to day 16 with an average of 30 eggs a day. During the following days (from day 23 to day 45), we found that the optimal fecundity was observed every second day and that 70.47% of the eggs laid during the entire life of the females were fertile. We also found that the fertility decreased significantly when the females were older than 38 days. As result, the age of hoverfly females influence significantly on their reproduction, with suggesting that hoverfly females from 2 to 5 weeks old are important agents for biological control programs. PMID- 18399481 TI - Ecology of aphidophagous predators in pomegranate ecosystem in India. AB - The aphid, Aphis punicae Passerini (Homoptera : Aphididae) is a serious pest attacking pomegranate (Punica granatum L.), an important semi arid fruit crop grown widely in most parts of the country. The major predators found preying on A. punicae in pomegranate ecosystem were Cheilomenes sexmaculata (Fabricius), Scymnus sp., Pseudaspidemerus circumflexo (Motsch.), Paragus serratus (Fabricius), Ischiodon scutellaris (Fabricius) and Chrysopa sp. The population dynamics and spatial distribution of these predators in an unsprayed pomegranate ecosystem were studied at Indian Institute of Horticultural Research, Bangalore (12 degrees 58' N; 77 degrees 35'E), India during 2000-2002. The predators were found to be distributed uniformly among different tree quadrants and followed the same distributional pattern of A. punicae during their peak in January and February. The predator density was relatively higher in lower canopies than upper canopies. The spatial distribution of predators showed aggregate distribution pattern at higher mean densities and exhibited regular or under-dispersed distribution at lower mean densities. The temporal distribution of aphidophagous predators on A. punicae showed two peaks one during January - February and second during August - September The population of predators started building up along with aphid population and reached maximum at high aphid densities and declined as the prey availability declined. This indicated that predators followed the same trend of their prey, A. punicae, showing a clear numerical response. PMID- 18399482 TI - Inventarisation of natural enemies in witloof chicory fields: preliminary results. AB - Due to residues on the witloof chicory heads the use of insecticides is forbidden during forcing. At this moment a suitable chemical control of the leafminer Napomyza cichorii is carried out in the field, based on the phenology and the population density and taking into account the economic threshold level. No chemical treatment is recognized against the root aphid Pemphigus bursarius. For monitoring these pests during the growing season water traps were placed on observation fields, scattered over the Belgian endive production area. The large numbers and the wide variety of other insects caught in the traps in recent years indicate that there is a potential of natural enemies. An inventarisation of the native enemies in witloof chicory fields was started recently using yellow coloured water traps and pitfall traps; at the same time field observations were done. In all the observation fields we found besides Carabidae and Staphilinidae also Coccinellidae and Syrphidae in large numbers. The numbers and the diversity of Hymenoptera in the water traps were very large. Apart from the field observations witloof chicory heads were investigated during harvest. Parasitized pupae were collected and kept in a climate room. Dacnusa leptogaster (Braconidae), a specific parasite of the witloof chicory fly, hatched. This parasite was also found in the water traps during the growing season. PMID- 18399483 TI - Perspectives for the biological control of Cameraria ohridella. AB - The horse chestnut leaf-miner, Cameraria ohridella Deschka et Dimic (Lepidoptera, Gracillariidae) is a serious invasive pest of Aesculus hippocastanum in Europe. The larvae of this species feed on leaf parenchyma and can reduce the tree growth. We studied the impact of parasitoids on C. ohridella in the Czech Republic and also searched for entomopathogenic fungi associated with this pest. The results showed that the rate of parasitism varied between 5% and 15% in most cases. The most parasitized stages of C. ohridella were spinning stages and especially pupae. The most abundant parasitoid species were Minotetrastichus frontalis, Pnigalio sp. and Pediobius saulius (Hymenoptera, Eulophidae). All species are polyphagous. Using the Galleria-bait method we isolated many strains of entomopathogenic fungi. Dominant species were Paecilomyces fumosoroseus, Paecilomyces farinosus and Beauveria bassiana. The perspectives of fungal bioagents in control of C. ohridella is discussed. PMID- 18399484 TI - The occurrence, population dynamics and natural regulation of abundance of Phyllonorycter robiniella (Clem.) in Krakow area. AB - This work covers the four years (2002-2005) observations on the occurrence and numerousness of Phyllonorycter robiniella (Clem.) and their natural regulation by parasitoid complex in Krakow area. The highest infestation by P. robiniella was found in year 2003 when almost 100% of Leaves and 90% of leaflets was infested. During the vegetation period the density of the observed mines increased and reached the maximum in August in all years of observations. The most important factor decreasing the number of P. robiniella was parasitization. The parasitization ranged between 18% and 27% depends on year of observation. Generally the highest parasitization was noted in the end of growing season, in September after the maximum of pest abundance. The most common parasitoids were Hymenoptera from the family Eulophidae (Chalcidoidea). PMID- 18399485 TI - Control of Cydia pomonella L. and Cydia molesta (Busck) (Lepidoptera Tortricidae) in pome-fruit orchards with Ecodian sex pheromone dispensers. AB - A mating disruption approach using high densities of pheromone dispensers, has been recently proposed for codling moth, Cydia pomonella (L.), and oriental fruit moth, Cydia molesta (Busck.), (Lepidoptera Tortricidae), control. Ecodian Star dispensers, made of low-cost biodegradable material and easy to apply, were formulated with 10 mg of codlemone (E8,E10-12OH) and 10 mg of grapamone (Z8-12OH) and placed at a rate of 1,400-2,000 dispensers/ha. The pheromone release rates from new and field aged dispensers were evaluated by hexane extraction of the residual attractant (indirectly) and gas-chromatographic analysis. The release rate of field-aged dispensers decreased over time with a good linearity; they released a significant amount of synthetic sex pheromones over the entire season. Dispensers elicited close-range approaches of codling moth males in wind tunnel irrespective of their age. Field trials carried out from 2003 to 2004 confirmed the efficacy of Ecodian Star dispensers for codling moth and oriental fruit moth control, regardless the size of the treated area. Our results demonstrate that Ecodian dispensers achieved a good level of activity and longevity over the season. The potential of this strategy for the control of the moths is discussed. PMID- 18399486 TI - Optical brightener Tinopal C1101 as an ultraviolet protectant for a nucleopolyhedrovirus. AB - Baculoviruses are arthropod-specific viruses currently used as biological insecticides in several countries of the world. One important factor limiting the efficacy of these bioinsecticides is related to the inactivation of the virus by solar ultraviolet (UV) radiation. This has motivated studies focused on the use of optical brightener compounds that can protect the virus from UV inactivation. The effects of the optical brightener Tinopal C1101 (ethenedyl benzenesulfonic derivative) on the persistance of a nucleopolyhedrovirus (SfMNPV) was quantified in third instar Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), the principal pest of maize in the Americas. For this, laboratory studies were performed to determine the relationship between virus concentrations and radiation-caused inactivation of SfMNPV alone or in mixtures with 1.25 or 0.1% (vol/vol) Tinopal C1101 using a UV system [radiation was provided by a 15 W UV tube (emission maxima at 312 nm)]. SfMNPV alone was more sensitive to UV than in mixtures with 1.25 or 0.1% Tinopal C1101. For instance, at concentrations of 2.22 x 10(8) and 2.75 x 10(6) occlusion bodies (OBs)/ml, SfMNPV alone-caused mortality was reduced from 88 and 44% (without UV radiation) to 0%, in both cases, after 30 and 15 min exposure to the UV tube, respectively. In contrast, the incorporation of 1.25% Tinopal C1101 into the SfMNPV inoculum at 2.22 x 10(8) and 2.75 x 10(6) OBs/ml caused a mortality of 86.6 and 96.6% after 240 min, respectively. UV caused inactivation of SfMNPV was directly related to the optical brightener concentration. We conclude that optical brighteners are likely to represent valuable components of nucleopolyhedrovirus formulations. PMID- 18399487 TI - Interaction effects between Beauveria bassiana and imidacloprid against Thrips tabaci (Thysanoptera: Thripidae). AB - The onion thrips, Thrips tabaci Lindeman is a major pest of many horticultural crops in many parts of the world. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin and imidacloprid are used for the control of the onion thrips. The potential interactions between the fungus and the chemical against the onion thrips adults were evaluated in a laboratory and two greenhouse bioassays. The laboratory bioassay was a leaf dip bioassay for a mixture of B. bassiana and imidacloprid. The first greenhouse bioassay was a topical application of a mixture of the fungus and the chemical while the second bioassay was a drenching application of imidacloprid and a topical application of B. bassiana. In all bioassays, the combined application resulted in higher percentage mortalities than either B. bassiana or imidacloprid alone. In the laboratory bioassay, the highest mortality of 97% was achieved by using B. bassiana at the field rate with imidacloprid at 1/10th the field rate relative to 88%, 94%, 21% for B. bassiana alone, imidacloprid alone and the control, respectively. In the first greenhouse bioassay, the highest mortality of 80% resulted from the combined application of B. bassiana and imidacloprid at field rates compared with 55%, 75%, and 22% for B. bassiana alone, imidacloprid alone and the control, respectively. In the second greenhouse bioassay, the combined application of both control agents at field rates resulted in 85% mortality compared with 52%, 83%, and 18% for B. bassiana alone, imidacloprid alone and the control, respectively. The interaction effect between the two control agents was additive except when B. bassiana at 1/2 field rate was mixed with imidacloprid at 1/10th field rate in the laboratory bioassay, where the interaction was antagonistic. The above results indicate that combining imidacloprid with B. bassiana might reduce the rate of the insecticide application while enhancing the efficacy of the biological control agent. PMID- 18399488 TI - Sublethal effects of two neurotoxican insecticides on Araneus pratensis (Araneae: Araneidae). AB - Spiders are important predators of several agricultural pests and they play an important role as indicators of ecosystem disturb. In Argentina, soybean crop has increased from the introduction of transgenic soybean resistant to glyphosate. This expansion produced an increase in the use of conventional and non-selective pesticides to control soybean pests. The objective of this work was to evaluate the side effects of subletal concentrations of two neurotoxican insecticides with a different mode of action: endosulfan (Glex, 35%, 25 mg/l a.i.) and spinosad (Tracer, 48%, 30 and 3 mg/l a.i) on Araneus pratensis. The insecticides were applied by ingestion of the treated prey (Musca domestica), and the effects on mortality, prey consumption, web building, mating, ootheca construction and fecundity were determined. Spinosad (30 mg/l a.i.) produced higher mortality than endosulfan (25 mg/l a.i.). Tremors and non-coordinated movements were observed in this treatment. The prey consumption was significantly reduced by the two insecticides (approximately 40% lower than control). The spider web building was significantly affected by the two insecticides, but spinosad had a greater effect. Though mating was not affected by both pesticides, abnormal oothecas and dehydrated eggs were observed. This work reports that sublethal concentrations representing approximately from 25 to 2.5% of the maximum field recommended concentrations (105 and 120 mg/l a.i., respectively) showed negative effects on A. pratensis. The consequences of these effects on role of A. pratensis as a natural mortality factor of soybean pests are discussed. PMID- 18399489 TI - Effect of conventional and biorational insecticides on larvae of Chrysoperla externa. PMID- 18399490 TI - An approach for IPM program to control sucking pests infesting garden bean plants (Phaseolus vulgaris L.) in Egypt. AB - An approach to eatablish IPM program for some sucking pests infesting garden beans plants was carried out in two successive seasons of 2004 and 2005 under field conditions in Alexandria, Egypt. The sucking pests included the aphid, Aphis craccivora Koch (Homoptera: Aphididae), the white fly, Bemisia tabaci Genn. (Homoptera: Aleyroidae) and the spider mite, Tetranychus urticae Koch Pymetrozine, thiamethoxam, carbosulfan, abamectin, azadirachtin and the detergent (Masrol 410) were tested for their efficacy in this study. Thiamethoxam, carbosulfan, and pymetrozine proved to be highly effective against Aphis craccivora Koch, while abamectin, azadirachtin, and the detergent (Masrol 410) provided a moderate effect. Moreover, thiamethoxam, pymetrozine, and abamectin showed remarkable efficacy against adults and immature stages of the whitefly, Bemisia tabaci. However, azadirachtin provided a moderate effect, while carbosulfan and detergent (Masrol 410) had no effect altogether on whiteflies. In addition, abamectin induced the highest toxicity effect against the red spider mite, Teteranychus urticae followed by azadirachtin and the detergent that provided moderate effects. IPM program could be achieved by using abamectin, pymetrozine and azadirachtin in a sequence pattern throughout the whole season. Furthermore, the performance of the used agrochemicals on certain nutritional elements of beans pods vr. Branco , i.e. nitrogen, phosphorus, potassium, iron, and magnesium were also studied with varied results. PMID- 18399491 TI - Abamectin, pymetrozine and azadirachtin sequence as a unique solution to control the leafminer Liriomyza trifolii (Burgess) (Diptera: Agromyzidae) infesting garden beans (Phaseolus vulgaris L.) in Egypt. AB - Field trails were conducted to determine the performance of three different sequences as a unique solution for the control of the leaf miner Liriomyza trifolii (Burgess) (Diptera: Agromyzidae) infesting garden beans (Phaseolus vulgaris L.) during the two successive seasons of 2004 and 2005. Furthermore, during the evaluation period, the side effect against the ectoparasite Diglyphus isaea (Walker) (Hymenoptera: Eulophidae) was put into consideration. Meanwhile, the comparative evaluation of the pesticides alone showed that abamectin and azadirachtin were highly effective against Liriomyza trifolii, while carbosulfan, pymetrozine and thiamethoxam provided to be of a moderate effect. Moreover, carbosulfan showed harmful effect to the larvae of the ectoparasite Diglyphus isaea (Walker), while abamectin and azadirachtin gave a moderate effect. Thiamethoxam and the the detergent (Masrol 410) had slight effect in this respect. The highly effective sequence among the sequences was abamectin, pymetrozine and azadirachtin, against Liriomyza trifolii (Burgess), with slight harmful effect on Diglyphus isaea (Walker). However the sequence of azadirachtin, pymetrozine and abamectin had a moderate effect on Liriomyza trifolii (Burgess) and exhibited a slight toxic effect on Diglyphus isaea (Walker). In contrast, the sequence of carbosulfan, thiamethoxam and pymetrozine was the least effective and represented a slight effect on Diglyphus isaea (Walker). From this study, it was concluded that abamectin, pymetrozine and azadirachtin sequence has proved to be a unique solution for the control of the leaf miner Liriomyza trifolii (Burgess) infesting garden beans (Phaseolus vulgaris L.) in Egypt. PMID- 18399492 TI - Effects of seed coatings with thiamethoxam on germination and flea beetle control in flax. AB - Flea beetles damage the seedlings before and after emergence and later the top of the plants as well, resulting in forking of the stem. This is unacceptable for the flax fibre industry, since it influences fibre quality negatively. Five field trials were carried out in 2001, 2003 and 2004 to assess the control of the flax flea beetles Longitarsus parvulus (Payk) and the large flax flea beetle Aphthona euphorbiae (Schrank) by film coating the seeds with thiamethoxam in fibre flax crops (Linum usitatissimum L.). Seed treatments were compared with untreated seeds and standard post emergence sprays with deltamethrin or parathion-methyl. Film coatings of the seeds with thiamethoxam formulated as 280 g/l, 350 g/l and 600 g/l were used. Thiamethoxam 350 g/l at rates of 9.1 and 18.2 g a.i./kg seed resulted in phytotoxicity, shown by a decrease of emergence. Application of thiamethoxam 350 g/l and 600 g/l at rates of 1.1 g a.i./kg seed and higher resulted in excellent flea beetle control. Application of thiamethoxam 280 g/l at a rate of 1.1 g a.i./kg seed showed no decrease of attack in comparison with the standard spray treatment. Seed treatment with thiamethoxam 600 g/l at 0.6 g a.i./kg showed insufficient protection. PMID- 18399493 TI - Pheromone trap and population model-based control of the codling moth, Cydia pomonella L., in Romanian apple culture. AB - Like in most parts of the world, the codling moth, Cydia pomonello (L.), is one of the most important pests in apple orchards in the Romanian Banat region. There, the control of this pest is often inadequate and the damage and economic losses are therefore enormous. This lack of control is caused by structural problems such as the absence of an advanced distribution network for phytopharmaceutical products, obsolete spraying equipment and the insufficient exchange of knowledge and expertise of those products and the pest itself between the local research facilities and the apple growers. The purpose of this research project is rather demonstrative, because a change in the Romanian pome culture's attitude is urgently needed. Pheromone trapping and the computerized population model RIMpro Cydia, which predicts important phenological events such as adult flight and egg hatch, were used to determine the optimal spray timing. These modern pest management techniques would not only minimize pressure on the environment, but also on the income of the Romanian apple grower, for whom phytopharmaceuticals are a high-priced investment. In July 2006 a comparative study between the supervised pest control program using Calypso, Runner and Reldan, all easily obtainable and commercially available insecticides in the region, and a conventional Romanian management programme was conducted. The control plot and the two test plots were all three located in a high codling moth pressure orchard in Timisoara. In the control plot 9,44 % of the fruits were damaged by codling moth larvae. In the plot containing the trees that were treated according to the traditional Romanian management programme 5,56 % damage was recorded, while the supervised management plot showed only 1,1% damage. Additionally damage was measured in another orchard in Periam, about 60 km away from the other plots. There, an intensive control program using Reldan, Calypso, Actare, Karate and Insegar was applied and damage measured only 0.2%. While at first glance that approach seems most efficient, it has to be noticed that it used twice as much insecticide treatments as were used in the supervised control program. Furthermore, 1 % damage is suggested by most references as being the economical threshold for treatments. The results of this comparative study clearly show that supervised codling moth control proofs to be the most profitable. Starting from August the same year, all of the control programmes in Timisoara were stopped. Therefore the second and possibly the third generation of codling moth larvae could develop freely. In the plot which was earlier sprayed according to the warnings of RIMpro Cydia and the pheromone trap this resulted in 27.5% damaged fruits during harvest in September. In the control plot damage reached 60.95%. These results highlight the necessity of the development of an efficient codling moth control program. Once that program is fine-tuned it should be presented to the local apple growers in order to give the Romanian pome culture a boost. PMID- 18399494 TI - Biological performance of quercetin on the cotton leaf-worm larvae, Spodoptera littoralis Boisd. (Lep., Noctuidae) and prevailing natural enemies in the Egyptian cotton fields. AB - The Egyptian cotton (Gossypium barbadense L.) accounts for 65 % of the world production of long stable cultivars. Taking into consideration the competition of other cotton producing countries, it should be of great importance to control pests, which attack the cotton plants to improve the yield and its quality. The main objective of this study is to develop new approaches for the management of the cotton leafworm Spodoptera littoralis Boisd. within an IPM program, that include synthetic insecticides rationalization, and maximiziation the role of the biological control agents. Sunflower plants Helianthus annuus (Asterales: Asteraceae) raised in rows surrounding plots of cotton were used as trap plants to attract some biological agents, which subsequently lead to check the build-up of the cotton leafworm population. This scientific phenomenon was attributed to the main chemical constituent of sunflower plants, which has been proved to be the polyhydroxy flavone "quercetin". Field data of the two successive seasons 2004 and 2005 revealed that: (a) the total number of insect predators, Coccinella undecimpunctata, Paederus alfierli, Chrysopa vulgaris, Orius laevigatus, Scymnus synacus, and true spiders in the cotton plots surrounded by either one or two rows of sunflower plants significantly exceeded the corresponding numbers in the cotton plots without sunflower plants., (b) the least number of cotton leafworm Spodoptera littorolis larvae infestation was recorded simultaneously in the cotton plots surrounded by sunflower plants. Moreover, laboratory studies assured the antifeeding properties of quercetin against the 4th instar larvae of Spodoptera littoralis. Quercetin at a concentration rate of 4000 ppm, showed abnormal behaviour represented in feeding stop, growth inhibition and development retardation. Deformation of pupae, moths, and reduction up to 50% in egg laying was also noticed after quercetin application to the larvae. PMID- 18399495 TI - In vivo effect of ACE inhibiting in mealworms on ovarian composition and ecdysteroid amounts. AB - Captopril is an inhibitor of angiotensin converting enzyme (ACE) used against arterial hypertension in mammals. The aim of this study was to evaluate captopril in the mealworm Tenebio molitor (Coleoptera: Tenebrionidae). The drug was diluted in acetone and administered topically (10 microg/3microl) on newly emerged adult females. The biochemical composition (proteins, lipids, carbohydrates, nucleic acids) and the ecdysteroid amounts of ovaries were determined at day 4 following treatment corresponding to the end of the vitellogenesis process. Biochemical analyses revealed that captopril applied in vivo reduced both total protein, lipid and carbohydrate amounts of ovaries, respectively. In addition, measurement of ecdysteroids by an enzyme-immunoassay indicated a significant decrease in ovarian ecdysteroid amounts. As a consequence, the reduction of reproductive capacity is possibly related to ecdysteroid production. Further experiments are needed to determine the action mechanism of captopril and the functions of ACE in insects. PMID- 18399496 TI - Effect of a fungal lectin from Xerocomus chrysenteron (XCL) on the biological parameters of aphids. AB - Aphids are important pests of crop plants in Europe. Increasing resistance of aphids to insecticides and their side effects on the environment and non target organism's including human's stimulated research on alterative methods of aphid control, including the use of entomotoxic proteins. Lectins are carbohydrate binding proteins that are widely distributed in nature; they have been isolated from microorganisms, fungi, plants and animals. Several of these proteins were tested for their potential biocide effect on plenty of pests. A fungal lectin, namely Xerocomus Chrysenteron lectin (XCL) was previously purified and was shown to be toxic for several pests including aphids. XCL was clearly the most toxic lectin against M. persicae. In this work, bioassays using artificial diets incorporating a broad range of XCL concentrations (from 10 microg x ml(-1) to 5000 microg x ml(-1)) were developed to assess the negative effects of XCL on the biological parameters (development duration, weight and fecundity) of M. persicae a polyphagous aphid found on more than 400 host plant species and transmitting more than 100 viral diseases. A significant mortality of aphids was observed, corresponding to the LC50 and LC90 of 0, 46 and 6, 02 mg/ml respectively after 24hrs. Significant differences of M. persicae weight, development duration and fecundity (P < 0.05) was observed between the tested XCL concentrations. Conavalia ensifomris lectin (ConA) was included as lectin reference on the bioassay experiments and was shown to be less toxic and induced lower negative changes in M. persicae biological parameters when compared with XCL. PMID- 18399497 TI - Analysis of lectin concentrations in different Rhizoctonia solani strains. AB - Lectins are carbohydrate-binding proteins that contain at least one carbohydrate binding domain which can bind to a specific mono- or oligosaccharide. These proteins are widely distributed in plants. However, over the last decade evidence is accumulating that lectins occur also in numerous fungi belonging to both the Ascomycota and Basiodiomycota. Rhizoctonia solani is known to be an important pathogen to a wide range of host plants. In this study, isolates of R. solani from different anastomosis groups have been screened for the presence of lectin using agglutination assays to detect and quantitate lectin activity. The evaluation included determination of the lectin content in mycelium as well as in sclerotia. The amount of lectin in the sclerotia was higher than in the mycelium of the same strains. The R. solani strains with the highest amounts of lectin have been selected for cultivation, extraction and purification of the lectin. PMID- 18399498 TI - First results on the insecticidal action of saponins. AB - In the search for new, natural insecticides, numerous scientists are currently trying to obtain useful compound from plants. A possibly interesting class of molecules are the saponins, a group of steroidal or triterpenoidal secondary plant metabolites with divergent biological activities. In this study, we investigated the activity of saponins against living caterpillars Spodoptera littoralis) and aphids (Acyrthosiphon pisum) via treatment on artificial diets containing different concentrations of saponins. We conclude that saponins have insecticidal activity, causing mortality and/or growth inhibition in the tested insects, although from our experiments the mode of action could not be identified. PMID- 18399499 TI - Autotropic in vitro model system for the in-depth study between nematodes and Rhizobium spp. PMID- 18399500 TI - Root vs pod infection by root-knot nematodes on aflatoxin contamination of peanut. AB - Aflatoxins are potent carcinogens produced by some Aspergillus spp. Infection of peanut (Arachis hypogaea) by root-knot nematodes (Meloidogyne arenaria) can lead to an increase in aflatoxin contamination of kernels when the plants are subjected to drought stress during pod maturation. It is not clear whether the increased aflatoxin contamination is primarily due to greater invasion of the galled pods by toxigenic Aspergillus spp. or whether root galling is also involved. Our objective was to determine the contribution of root and pod galling caused by root-knot nematodes to the increase in aflatoxin contamination in peanut. Two greenhouse experiments were conducted in which pods and roots were physically separated. Pod set was restricted to soil-filled pans (41 cm dia. x 10 cm depth), while the roots grew underneath the pan into a pot. The experiments had a factorial arrangement of treatments: pod zone with and without nematodes, and root zone with and without nematodes. In Experiment 1, 5000 eggs of M. arenaria were added to the root zone14 days after planting (DAP) and 8000 eggs were added to the pod zone 60 and 80 DAP. In Experiment 2, 3000 eggs were added to the root zone 30 DAP and 8000 eggs were added to the pod zone every week starting 60 DAP. The four treatment combinations were replicated 10 to 13 times. Conidia of Aspergillus flavus/A. parasiticus was added to the soil surface (pods zone) at mid bloom. Plants were subjected to drought stress 40 days before harvest. In Experiment 1, adding nematodes to the pod zone had no effect on aflatoxin concentrations in the peanut kernel. However, the lack of an effect may have been to due to the low occurrence of galling on the hulls. In pots where nematodes were added to the root zone, 50 to 80% of the root system was galled. Adding nematodes to the root zone increased aflatoxin concentrations in the peanut kernels from 34 ppb in the control to 71 ppb. In Experiment 2, there was heavy pod galling with galls present on 53% of the pods. Adding nematodes to the pod zone increased aflatoxin concentrations in the kernels from 19 ppb in the control to 572 ppb. Based on the results of the two experiments, it appears that infection of either the roots or pods by M. arenaria can lead to greater aflatoxin contamination of peanut kernels. PMID- 18399501 TI - Response of five lettuce cultivars to root-knot nematode, Meloidogyne incognita. AB - The root-knot nematode, Meloidogyne incognito (Kofoid et White) Chitwood is an important pathogen of vegetables. Five commercial cultivars of lettuce (Lactuca sativa L.) were evaluated under greenhouse conditions for resistance to Meloidogyne incognita, Benguet population. Plants were inoculated with 1000 eggs collected from 'Apollo' tomato (Lycopersicon esculentum) roots. The degree of galling and number of egg masses were assessed 4 and 8 weeks after inoculation. Host plant response was classified as immune, highly resistant, resistant, moderately resistant, intermediate, moderately susceptible, and highly susceptible based on the resistance index of Kouame et at., 1998 [RI = (gall2 + egg2)]. Inoculation of 1000 eggs/plant significantly affected the growth and yield of the five lettuce cultivars 4 and 8 weeks after inoculation. A significant interaction was observed between treatment and cultivar during the two evaluation periods in terms of marketable and non-marketable yield, plant height, root weight, number of galls and number of egg masses. A reduction in growth and yield was observed in the cultivars Ballon, Lollo Rosa and Red Wave. Significant differences were noted in the number of galls and egg masses among the different cultivars tested. The highest average number of galls was obtained from the cultivars Red Wave, Ballon and Lollo Rosa. Cultivar Ballon had the highest average number of recovered nematode while Gilaben had the lowest with 15 and 4 per roots, respectively after 4 weeks inoculation. After 8 weeks, nematode was highest in cultivar Red Wave (615) and lowest in Great Lakes (70). Based on the host response, cultivars Great Lakes and Gilaben were rated highly resistant and resistant, respectively, while Red Wave, Ballon and Lollo Rosa were rated intermediate. PMID- 18399502 TI - AMF-induced bioprotection against migratory plant-parasitic nematodes: which mechanisms are responsible? PMID- 18399503 TI - Management of the potato cyst nematode (Globodera pallida) with bio fumigants/stimulants. AB - Field trials evaluated the effect of four plant-based bio-fumigants/stimulants on population levels of G. pallida and the resulting potato yields and quality. Three formulations contained seaweed biostimulants (Algifol, Nutridip and Metastim) and one bio-fumigant containing mustard and chilli pepper extracts (Dazitol). These were compared with the fumigant nematicide Nemathorin and untreated control plots. The effect of G. pallida on growing potato crops was assessed by recording haulm characteristics which indicated that the nematicide treatment gave most protection. Levels of PCN juveniles and migratory nematodes were assessed during the trial. Plots treated with Nemathorin and Dazitol had fewest PCN, whilst the highest number of migratory nematodes occurred in fallow plots. Sixteen weeks after planting the nematicide treatment produced highest yield and tuber numbers. Dazitol treatment produced a lower yield but the largest tubers. PMID- 18399504 TI - The global importance of the cereal cyst nematode (Heterodera spp.) on wheat and international approaches to its control. AB - The Cereal Cyst Nematodes (CCNs) are a group of several closely related species which have been documented to cause economic yield loss on rainfed wheat production systems in several part of the world including North Africa, West Asia, China, India, Australia, America and several countries in Europe. The most commonly reported species is Heterodera avenae, however there are at least two other species H. filipjevi and H. latipons are implicated. It is well appreciated that plants under water and nutrient stress suffer greater yield loss. Control of CCNs requires maintaining nematode populations below economic thresholds. Chemicals are not environmentally sustainable or economic and the major emphasis on control has been with host genetic resistance applied with other integrated pest managent options. Unfortunately due to the number of species and pathotype variation genetic control of Cereal Cyst Nematode with plant resistance is complex. Turkey is one of the top ten wheat producers in the world and has identified these nematode as a major biotic constraint in their rainfed wheat systems. In 2001 a new joint intiative was established between CIMMYT International, the Turkish Ministry of Agriculture and (Ukurova University in Adana to understand i) the distribution of cereal nematodes on wheat; ii) assess the economic importance and improve our understanding of the population dynamics iii) culture, screen and assess known sources of resistance and identify new sources to both groups of nematodes; iv) integrate new sources of resistance into bread wheat cultivars for Turkey and International germplasm using conventional and molecular tools; v) investigate other integrated control options such as rotation and different wheat management strategies and finally vi) capacity build scientists to work in this important area. Some highlights of this work will be presented and the newly formed ICCNI - International Cereal Cyst Nematode Initative introduced. PMID- 18399505 TI - Optimization of metam sodium application by rotary spading injection. AB - In The Netherlands the only chemical alternative for methyl bromide permitted is an application of metam-sodium (MS) with the active ingredient methyl isothiocyanate (MIT). After introduction of a new application method with 'rotary spading injection' legislation restricted the application of MS in 1993 to once in four years and since 2001 once in five years. Efficacy after injection of metam sodium at 10 cm depth and rotary spading a 25 cm soil layer was much better than with shank injection at 19 cm depth with a poor efficacy in the top soil layer. Legislation for arable crops allows a dosage of 300 l Monam (510 g MS/l) per ha. For vegetables, fruits, bulbs and weeds dosages permitted are 600 to 750 l Monam per ha. For an optimal disinfestation result the combination of required dosage and injection depth and rotovation depth had to be established. In a field experiment two dosages Monam were tested for efficacy against Meloidogyne fallax with rotary spading injection at varying injection depths. 300 l Monam per ha was compared with 600 l Monam per ha, both applied at 14 cm injection depth and distributed over a 28 cm soil layer. Another treatment was the application of 600 l Monam per ha, injected at 20 cm depth and rotovated through 40 cm of soil. A carrot crop was sown three weeks after disinfestation and quantity and quality of carrot yield was assessed. Three weeks after application Meloidogyne fallax population was reduced in the furrow with 99% and at 30-50 cm depth with 96% by both applications of 600 l Monam. Efficacy of 300 l Monam was significantly less than 600 l. Nematode population levels after carrot crop stayed lower in the furrow after 600 l in comparison with 300 l Monam. Gross carrot yield was significantly higher after 600 l than after 300 l Monam application at 14 cm depth. Although on this heavy infested field it was not possible to grow carrots without symptoms, after 600 l Monam the percentage of carrots without root-knot symptoms (net yield) was statistically higher than after 300 l Monam. PMID- 18399506 TI - Tolerance of Steinernema feltiae (Rhabditida: Steinernematidae) to agrochemical products. PMID- 18399507 TI - Pratylenchus goodeyi, a species-complex? PMID- 18399508 TI - Evaluation of soil biodesinfestation with crop and garden residues in the control of root-knot nematodes populations. AB - Fresh crop and garden residues were applied both under laboratory conditions and in commercial greenhouse in order to asses their effect on soil nematodes populations and soil fertility. In the laboratory experiments, dosages of 5 to 20 g of cabbage residues, chicken manure, cabbage residues+chicken manure, grass+chicken manure, as well as leaves and stems of orange tree, pine tree, oleander, olive tree, palm tree and boxwood were mixed with 500 g soil having root-knot nematodes (Meloidogyne incognita) and soil moisture was adjusted at field capacity. A control treatment without residues was also included. The mixtures were kept into plastic bags, with four replications, and the bags were incubated for four weeks at 30 degrees C, when nematological and soil fertility analyses were carried out. In general, all these materials significantly (P < 0.05) reduced M. incognita populations and increased saprophagous nematodes, with slight effects on soil fertility except for the K increase with residues application. Tomato plants susceptible to M. incognita were planted in pots with 300 cm3 of the treated soils and kept for five weeks in a growth chamber (24 +/- 1 degrees C, 14 hours light), when root galling indices were evaluated. Most materials applied reduced root galling indices as regards to the control. In the greenhouse experiment, cabbage residues, cabbage residues+chicken manure, grass+chicken manure and grass+cabbage residues were applied to the soil and covered with a polyethylene sheet for 5 weeks. A cabbage residues:chicken manure treatment and a control (not-amended) treatment, without polyethylene, were also included. At the end of the experiment, the nematological analysis showed that all materials successfully controlled M. incognita populations, reaching 86-100% mortality with organic amendments vs. 6% for the control. After the greenhouse biodesinfestation experiment, a tomato crop was grown for one month, when root galling indices were determined. All materials significantly reduced this value from 4.75 in the control to 1.0-2.25 with the organic amendments, except for the cabbage residues+chicken manure treatment without polyethylene (index = 4.0). Our results show that fresh crop and garden residues successfully reduced M. incognita populations and root galling indices when applied with polyethylene covers, having good potential to be considered in integrated management programs. PMID- 18399509 TI - D-(+)-iso-methanocarbathymidine: a high-affinity substrate for herpes simplex virus 1 thymidine kinase. AB - The stereoselective syntheses of the (+)-D and (-)-L enantiomers of iso methanocarbathymidine (iso-MCT) was achieved through two independent linear approaches that converged on two antipodal enantiomers, common to a key precursor used in the synthesis of racemic iso-MCT. In the study reported herein we identified (+)-3 [D-(+)-iso-MCT] as the active enantiomer that was exclusively recognized by the herpes simplex virus 1 thymidine kinase (HSV1-tk), as was predicted by molecular modeling. For this purpose, a human osteosarcoma (HOS) cell line modified to contain and express HSV1-tk from herpes simplex virus 1 (HSV1) was used to determine the cytotoxicity of the compounds by an assay that measures the level of ATP in the cells. The work demonstrates that changes in the substitution pattern of rigid bicyclo[3.1.0]hexane nucleosides, which, relative to normal nucleosides, appear unconventional, can lead to the spatial optimization of pharmacophores and vastly improved substrate recognition. PMID- 18399510 TI - Varied active-site constraints in the klenow fragment of E. coli DNA polymerase I and the lesion-bypass Dbh DNA polymerase. AB - We report on comparative pre-steady-state kinetic analyses of exonuclease deficient Escherichia coli DNA polymerase I (Klenow fragment, KF-) and the archaeal Y-family DinB homologue (Dbh) of Sulfolobus solfataricus. We used size augmented sugar-modified thymidine-5'-triphosphate (T(R)TP) analogues to test the effects of steric constraints in the active sites of the polymerases. These nucleotides serve as models for study of DNA polymerases exhibiting both relatively high and low intrinsic selectivity. Substitution of a hydrogen atom at the 4'-position in the nucleotide analogue by a methyl group reduces the maximum rate of nucleotide incorporation by about 40-fold for KF- and about twelve fold for Dbh. Increasing the size to an ethyl group leads to a further twofold reduction in the rates of incorporation for both enzymes. Interestingly, the affinity of KF- for the modified nucleotides is only marginally affected, which would indicate no discrimination during the binding step. Dbh even has a higher affinity for the modified analogues than it does for the natural substrate. Misincorporation of either TTP or T(Me)TP opposite a G template causes a drastic decline in incorporation rates for both enzymes. At the same time, the binding affinities of KF- for these nucleotides drop by about 16- and fourfold, respectively, whereas Dbh shows only a twofold reduction. Available structural data for ternary complexes of relevant DNA polymerases indicate that both enzymes make close contacts with the sugar moiety of the dNTP. Thus, the varied proficiencies of the two enzymes in processing the size-augmented probes indicate varied flexibility of the enzymes' active sites and support the notion of active site tightness being a criterion for DNA polymerase selectivity. PMID- 18399511 TI - Novel contributions to the mechanism of the enantioselective hydrogenation of dimethyl itaconate with rhodium complexes. PMID- 18399512 TI - Synthesis, structure, and transport property of perfluorinated oligofluorenes. PMID- 18399513 TI - Highly enantioselective strecker reaction of ketoimines catalyzed by an organocatalyst from (S)-BINOL and L-prolinamide. PMID- 18399514 TI - alpha,alpha-Cyclopentaneglycine dipeptides capped with biaryls as tachykinin NK(2) receptor antagonists. AB - The NK(2) receptor belongs to the family of tachykinin neurotransmitters. It has been reported to be involved in several pathological conditions, and selective antagonists are potentially useful drugs for the treatment of asthma, irritable bowel syndrome, cystitis, and depression. Starting from in-house capped dipeptide libraries, we were able to identify a number of molecules with sub-nanomolar binding affinity for the hNK(2) receptor. All were characterized by a rigid core structure with a strong constraint induced by an alpha,alpha-cyclopentaneglycine fragment. Herein we report the further elaboration of three initial basic structures. The planar benzothiophene group was substituted with a series of biphenyl and heterobiphenyl moieties that are well tolerated in terms of receptor affinity. The new compounds also maintained good antagonist potency in an in vitro functional assay, and a number of them showed significant in vivo activity after intravenous administration in our guinea pig model. PMID- 18399515 TI - Influence of amino acid side chains on long-distance electron transfer in peptides: electron hopping via "stepping stones". PMID- 18399516 TI - A general approach to the size- and shape-controlled synthesis of platinum nanoparticles and their catalytic reduction of oxygen. PMID- 18399517 TI - Water-soluble nanocrystals through dual-interaction ligands. PMID- 18399518 TI - Large-scale synthesis of ultrathin Bi2S3 necklace nanowires. PMID- 18399519 TI - The versatile chemistry and noncentrosymmetric crystal structures of salt inclusion vanadate hybrids. PMID- 18399520 TI - Bacterial acyltransferases as an alternative for lipase-catalyzed acylation for the production of oleochemicals and fuels. AB - Bacterial acyltransferases are a new class of enzymes, and the first member was identified as WS/DGAT in Acinetobacter baylyi ADP1. Their unspecificity have been used in several biotechnological applications for lipid modification, a field that has been dominated by the use of lipases. Examples are the biosynthesis of jojoba-like wax esters and fatty-acid ethyl esters. In addition, these enzymes are also capable of synthesizing acylthioesters. Acyloxoesters and acylthioesters can thus be produced in vivo by whole-cell fermentations rather than in vitro in an enzyme reactor. In this Minireview, we focus on the biotechnological utilization of acyltransferases for the production of modified lipids from renewable resources. PMID- 18399521 TI - Tailored control and optimisation of the number of phosphonic acid termini on phosphorus-containing dendrimers for the ex-vivo activation of human monocytes. AB - The syntheses of a series of phosphonic acid-capped dendrimers is described. This collection is based on a unique set of dendritic structural parameters cyclo(triphosphazene) core, benzylhydrazone branches and phosphonic acid surface and was designed to study the influence of phosphonate (phosphonic acid) surface loading towards the activation of human monocytes ex vivo. Starting from the versatile hexachloro-cyclo(triphosphazene) N(3)P(3)Cl(6), six first-generation dendrimers were obtained, bearing one to six full branches, that lead to 4, 8, 12, 16, 20 and 24 phosphonate termini, respectively. The surface loading was also explored at the limit of dense packing by means of a first-generation dendrimer having a cyclo(tetraphosphazene) core and bearing 32 termini, and with a first generation dendrimer based on a AB(2)/CD(5) growing pattern and bearing 60 termini. Human monocyte activation by these dendrimers confirms the requirement of the whole dendritic structure for bioactivity and identifies the dendrimer bearing four branches, thus 16 phosphonate termini, as the most bioactive. PMID- 18399522 TI - Estimating the temperature dependence of peptide folding entropies and free enthalpies from total energies in molecular dynamics simulations. AB - The temperature dependence of thermodynamic quantities, such as heat capacity, entropy and free enthalpy, may be obtained by using well-known equations that relate these quantities to the enthalpy of the molecular system of interest at a range of temperatures. In turn, the enthalpy of a molecular system can be estimated from molecular dynamics simulations of an appropriate model. To demonstrate this, we have investigated the temperature dependence of the enthalpy, heat capacity, entropy and free enthalpy of a system that consists of a beta-heptapeptide in methanol and have used the statistical mechanics relationships to describe the thermodynamics of the folding/unfolding equilibrium of the peptide. The results illustrate the power of current molecular simulation force fields and techniques in establishing the link between thermodynamic quantities and conformational distributions. PMID- 18399523 TI - Controllable synthesis of nickel hydroxide and porous nickel oxide nanostructures with different morphologies. AB - Alpha-Ni(OH)(2) nanobelts, nanowires, short nanowires, and beta-Ni(OH)(2) nanoplates have been successfully prepared in high yields and purities by a convenient hydrothermal method under mild conditions from very simple systems composed only of NaOH, NiSO(4), and water. It has been found that the ratio of NaOH to NiSO(4) not only affects the morphology of the Ni(OH)(2) nanostructures, but also determines whether the product is of the alpha- or beta-crystal phase. A notable finding is that porous NiO nanobelts were produced after exposure of the Ni(OH)(2) products to an electron beam for several minutes during transmission electron microscopy (TEM) observations. Another unusual feature is that rectangular nanoplates with many gaps were obtained. Furthermore, porous NiO nanobelts, nanowires, and nanoplates could also be obtained by annealing the as prepared Ni(OH)(2) products. A sequence of dissolution, recrystallization, and oriented attachment-assisted self-assembly of nanowires into nanobelts is proposed as a plausible mechanistic interpretation for the formation of the observed structures. The method presented here possesses several advantages, including high yields, high purities, low cost, and environmental benignity. It might feasibly be scaled-up for industrial mass production. PMID- 18399524 TI - Ionic liquids of bis(alkylethylenediamine)silver(I) salts and the formation of silver(0) nanoparticles from the ionic liquid system. AB - We have prepared novel ionic liquids of bis(N-2 ethylhexylethylenediamine)silver(I) nitrate ([Ag(eth-hex-en)(2)]NO(3) and bis(N hexylethylenediamine)silver(I) hexafluorophosphate ([Ag(hex-en)(2)]PF(6)), which have transition points at -54 and -6 degrees C, respectively. Below these transition temperatures, both the silver complexes assume amorphous states, in which the extent of the vitrification is larger for the eth-hex-en complex than for the hex-en complex. The diffusion coefficients of both the complex cations, measured between 30 (or 35) and 70 degrees C, are largely dependent on temperature; the dependence is particularly large in the case of the eth-hex-en complex cation below 40 degrees C. Small-angle X-ray scattering studies showed that the bilayer structure of the metal complex is formed in the liquid state for both the silver complexes. A direct observation of the yellowish [Ag(eth-hex en)(2)]NO(3) liquid by transmission electron microscopy (TEM) indicates the presence of nanostructures, as a microemulsion, of less than 5 nm. Such structures were not clearly observed in the [Ag(hex-en)(2)]PF(6) liquid. Although the [Ag(eth-hex-en)(2)]NO(3) liquid is sparingly soluble in bulk water, it readily incorporates a small amount of water up to [water]/[metal complex] = 7:1. Homogeneous and uniformly sized silver(0) nanoparticles in water were created by the reduction of the [Ag(eth-hex-en)(2)]NO(3) liquid with aqueous NaBH(4), whereas silver(0) nanoparticles were not formed from the [Ag(hex-en)(2)]PF(6) liquid in the same way. PMID- 18399525 TI - Tandem cofacial stacks of porphyrin-phthalocyanine dyads through complementary coordination. AB - A novel straightforward methodology to organize discrete heterogeneous stacks of porphyrin and phthalocyanine employed an imidazolyl-to-zinc complementary coordination protocol for a Zn(II) phthalocyanine that contains an imidazolyl terminal with an ethynylporphyrin as a coplanar spacer. Structural elucidation was performed by means of size-exclusion chromatography, spectral titration, and NMR spectroscopy. The association constants for the complementary coordination of the heterogeneous slipped-cofacial tetrads reached extremely high values, in the order of 10(14) M(-1). Close contact of the porphyrin and phthalocyanine planes led to a strong shielding of the cofacial protons, which were split due to the slipped-cofacial heterogeneous environment. In variable-temperature NMR spectroscopy, the split signals remained in the aromatic region, a result suggesting structural robustness. Addition of trifluoroacetic acid dissociated the coordination structure to unify the split signals. The stacked tetrads showed unique electronic structures, such as strong exciton coupling and charge-transfer properties between the porphyrin and phthalocyanine units, which were modulated by the peripheral substituents of the phthalocyanine subunit and by the solvent. Interconversion between the coordination tetrad and the corresponding dyad was observed upon addition of an axial ligand. PMID- 18399526 TI - Total synthesis of (-)-balanol, all stereoisomers, their N-tosyl analogues, and fully protected ophiocordin: an easy route to hexahydroazepine cores from garner aldehydes. AB - Total syntheses of (-)-balanol and all of its stereoisomers starting from easily available Garner aldehydes are described. Diastereoselective Grignard reactions on Garner aldehydes and ring-closing metatheses are the key steps for the construction of hexahydroazepine subunits. The benzophenone subunits were constructed through coupling of suitably functionalized aromatic aldehyde and bromo components. The synthetic route constitutes a convenient and scalable reaction sequence to generate all of the stereoisomers of balanol. The methodology is explored further for the synthesis of N-tosyl analogues of balanol and of fully protected ophiocordin. PMID- 18399527 TI - A core-shell nanoparticle approach to photoreversible fluorescence modulation of a hydrophobic dye in aqueous media. AB - Amphiphilic core-shell nanoparticles containing spiropyran moieties have been prepared in aqueous media. The nanoparticles consist of hydrophilic and biocompatible poly(ethyleneimine) (PEI) chain segments, which serve as the shell, and a hydrophobic copolymer of methyl methacrylate (MMA), a spiropyran-linked methacrylate, and a cross-linker, which forms the core of the nanoparticles. A hydrophobic fluorescent dye based on the nitrobenzoxadiazolyl (NBD) group was introduced into the nanoparticles to form NBD-nanoparticle complexes in water. The nanoparticles not only greatly enhance the fluorescence emission of the hydrophobic dye NBD in aqueous media, probably by accommodating the dye molecules in the interface between the hydrophilic shells and the hydrophobic cores, but also modulate the fluorescence of the dye through intraparticle energy transfer. This biocompatible and photoresponsive nanoparticle complex may find applications in biological areas such as biological diagnosis, imaging, and detection. In addition, this nanoparticle approach will open up possibilities for the fluorescence modulation of other hydrophobic fluorophores in aqueous media. PMID- 18399528 TI - Reactivity of silole within a core-modified porphyrin environment: synthesis of 21-silaphlorin and its conversion to carbacorrole. AB - Condensation of 1,1-dimethyl-3,4-diphenyl-2,5-bis(p-tolylhydroxymethyl)silole with pyrrole and p-tolylaldehyde did not form the expected 21,21-dimethyl-2,3 diphenyl-5,10,15,20-tetra(p-tolyl)-21-silaporphyrin, but rather its reduced derivative, 21-silaphlorin, which contains a tetrahedrally hybridised C5 carbon atom. Attempts to trap 21-silaporphyrin resulted in the serendipitous discovery of a unique transformation of 21-silaphlorin into a non-aromatic isomer of 2,3 diphenyl-5,10,15,21-tetra(p-tolyl)-carbacorrole (iso-carbacorrole). This novel carbaporphyrinoid contains a cyclopentadiene ring embedded in a tripyrrolic framework. This transformation of 21-silaphlorin to iso-carbacorrole, carried out under oxidative conditions, involves extrusion of dimethylsilylene accompanied by migration of the C(meso)-(p-tolyl) unit to create a cyclopentadiene ring directly linked to the adjacent pyrrole through a tetrahedral carbon atom. Insertion of silver or copper ions into iso-carbacorrole gave two structurally related organometallic complexes of "true" carbacorrole in which the metal(III) ions are bound by three pyrrolic nitrogen atoms and a tetrahedrally hybridised C21 atom of the cyclopentadiene moiety. In the presence of oxygen, the silver(III) carbacorrole undergoes internal oxidation to 21-oxacorrole. The structure of silver(III) carbacorrole was determined by X-ray crystallography. The C21 atom was found to have a tetrahedral geometry. The Ag-C(sp(3)) (2.046(5) A) bond length is similar to that in silver(III) carbaporphyrinoids in which a trigonal carbon atom coordinates to the metal ion. Density functional theory was applied to model the molecular and electronic structure of 21-silaphlorin and feasible isomers of carbacorrole. The total energies (kcal mol(-1) vs. iso-carbacorrole), calculated at the B3LYP/6-31G(**)//B3LYP/6-31G(*) level for carbacorrole, iso carbacorrole, vacataporphyrin and cyclobutadienephlorin, demonstrate the energetic preference for iso-carbacorrole. PMID- 18399529 TI - High catalytic activities of artificial peroxidases based on supramolecular hydrogels that contain heme models. AB - Composed of a supramolecular hydrogel and a heme model compound, a new type of artificial peroxidase shows high catalytic activity in organic media. The activity of this new type of artificial enzyme is significantly higher than that of the heme model compounds alone. Changes in the distal substituents above the coordinated-metal centers of the model compounds directly modulate catalytic activity. This supramolecular-hydrogel-based artificial enzyme is most active in toluene, reaching about 90% of the nascent activity of horseradish peroxidase. Moreover, this study confirms that the incorporation of the heme models into the nanofibers of gelators accounts for most of the enhancement of catalytic activity. PMID- 18399530 TI - New M(3)N@C(2n) endohedral metallofullerene families (M=Nd, Pr, Ce; n=40-53): expanding the preferential templating of the C(88) cage and approaching the C(96) cage. AB - Three new families of trimetallic nitride template endohedral metallofullerenes (TNT EMFs), based on cerium, praseodymium, and neodymium clusters, were synthesized by vaporizing packed graphite rods in a conventional Kratschmer Huffman arc reactor. Each of these families of metallofullerenes was identified and characterized by mass spectroscopy, HPLC, UV/Vis-NIR spectroscopy, and cyclic voltammetry. The mass spectra and HPLC chromatograms show that these larger metallic clusters are preferentially encapsulated by a C(88) cage. When the size of the cluster is increased, the C(96) cage is progressively favored over the predominant C(88) cage. It is also observed that the smaller cages (C(80)-C(86)) almost disappear on going from neodymium to cerium endohedral metallofullerenes. The UV/Vis-NIR spectra and cyclic voltammograms confirm the low HOMO-LUMO gap and reversible electrochemistry of these M(3)N@C(88) metallofullerenes. PMID- 18399531 TI - Developing a new class of axial chiral phosphorus ligands: preparation and characterization of enantiopure atropisomeric phosphinines. AB - Both enantiomers of the first atropisomeric phosphinine (1) have been isolated by using analytical HPLC on a chiral stationary phase. The enrichment of one enantiomer and a subsequent investigation into its racemization kinetics revealed a barrier for internal rotation of DeltaG(298)(double dagger) = (109.5+/-0.5) kJ mol(-1), which is in excellent agreement with the theoretically predicted value of DeltaG(298)(double dagger) =116 kJ mol(-1). Further analysis with UV and circular dichroism spectroscopies and density functional theory calculations led to the determination and assignment of the absolute configurations of both enantiomers. These results are the basis for future investigations into this new class of axially chiral phosphinine-based ligands and their possible applications in asymmetric homogeneous catalysis. PMID- 18399532 TI - Efficient synthesis of methylenetetrahydrofurans and methylenepyrrolidines by formal [3+2] cycloadditions of propargyl substrates. AB - Recent developments concerning the synthesis of methylenetetrahydrofurans and methylenepyrrolidines by one-pot formal [3+2] cycloadditions involving propargylic (and allylic) alcohols and amines with electrophilic alkenes are described. The synthetic methods provide powerful tools to prepare highly functionalized oxygen- and nitrogen-containing five-membered ring systems. The reactions can be effectively promoted by base, base/transition metals, and Lewis acids, depending on the substrates. PMID- 18399533 TI - Superstructure-dependent optical and electrical properties of an unusual face-to face, pi-stacked, one-dimensional assembly of dehydrobenzo[12]annulene in the crystalline state. AB - To develop a novel pi-conjugated molecule-based supramolecular assembly, we designed and synthesized trisdehydrotribenzo[12]annulene ([12]DBA) derivative 2 with three carboxyl groups at the periphery. Recrystallization of 2 from DMSO gave a crystal of the solvate 23 DMSO. Crystallographic analysis revealed, to our surprise, that a face-to-face pi-stacked one-dimensional (1D) assembly of 2 was achieved and that the DMSO molecule played a significant role as a "structure dominant element" in the crystal. This is the first example of [12]DBA to stack completely orthogonal to the columnar axis. To reveal its superstructure dependent optical and electrical properties, 2 and its parent molecule 1, which crystallizes in a herringbone fashion, were subjected to fluorescence spectroscopic analysis and charge-carrier mobility measurements in crystalline states. The 1D stacked structure of 2 provides a red-shifted, broadened, weakened fluorescence profile (lambda(max) = 545 nm, phi(F) = 0.01), compared to 1 (lambda(max) = 491 nm, phi(F) = 0.12), due to strong interactions between the p orbitals of the stacked molecules. The charge-carrier mobility of the single crystal of 23 DMSO, as well as 1, was determined by flash photolysis time resolved microwave conductivity (FP-TRMC) measurements. The single crystal of 23 DMSO revealed significantly-anisotropic charge mobility (sigma(mu) = 1.5x10(-1) cm(2) V(-1) s(-1)) along the columnar axis (crystallographic c axis). This value is 12 times larger than that along the orthogonal axis (the a axis). PMID- 18399534 TI - Module extension of a non-ribosomal peptide synthetase of the glycopeptide antibiotic balhimycin produced by Amycolatopsis balhimycina. PMID- 18399535 TI - Differential effect of oxidative stress on the apoptosis of early and late passage human diploid fibroblasts: implication of heat shock protein 60. AB - Since an attenuated response to stress is a characteristic of senescence, a cellular senescence model was used to examine the mechanism of resistance against oxidative stress using human diploid fibroblasts (HDF). With increasing passage, the HDF showed increased production of reactive oxygen species (ROS). Late passage HDF were resistant to the lethal effects of oxidative stress, showing less cleavage of pro-caspase-3 and PARP than those of early ones. Since heat shock proteins (Hsps) are not only cytoprotective but also interfere with the apoptotic cascade, the expression patterns of Hsps during cellular senescence were next examined. Oxidative stress induced a decrease in the mitochondrial Hsp60 levels with a concomitant increase in the cytosolic Hsp60 levels in the early passage HDF, but not in late ones. To show that the resistance to oxidative stress is a specific effect of Hsp60, the levels of Hsp60 were knocked down by siRNA. As expected the Hsp60 knock-down cells were more resistant to oxidative stress. These findings show that Hsp60 is a key player in the resistance mechanism against oxidative stress and aging. PMID- 18399536 TI - Effect of pH on stability of recombinant botulinum serotype A vaccine in aqueous solution and during storage of freeze-dried formulations. AB - The purpose of this study was to evaluate the importance of prelyophilization solution pH on the stability of botulinum neurotoxin, serotype A (rBoNTA(H(c))). This understanding is of significant importance for proteins such as rBoNTA(H(c)), a potential constituent of a multivalent vaccine product. For multivalent vaccines it may be difficult to identify a liquid formulation satisfying the stability requirements for all constituent protein antigens. Consequently, a lyophilized multivalent vaccine formulation may be a more viable alternative. Therefore evaluating the effect of prelyophilization pH (may be suboptimal) on the stability of antigens such as rBoNTA(H(c)) during lyophilization/storage becomes important. We hypothesize that when rBoNTA(H(c)) is lyophilized from a suboptimal pH, using the appropriate stabilizers can provide adequate physicochemical stability during lyophilization and long-term storage. We identified pH 5 and 8 in which the protein was stable and unstable against aggregation. Excipients were identified that could stabilize rBoNTA(H(c)) during lyophilization and storage in a stable solution of pH 5. These excipients were 7.5% (w/v) trehalose and 2.5% (w/v) trehalose with 2.5% (w/v) HES, with and without 0.01% (w/v) polysorbate 20. In support of our hypothesis, these excipients were found to provide adequate physicochemical stability to rBoNTA(H(c)) during lyophilization/storage, when freeze-dried from a prelyophilized solution of pH 8. PMID- 18399537 TI - Validation of in vitro cell culture models of the blood-brain barrier: tightness characterization of two promising cell lines. AB - In the course of the validation of blood-brain barrier in vitro models the aim of this work was to characterize two promising continuous cell lines with regard to their tightness properties. PBMEC/C1-2 and ECV304 cells were cultured in several media with different compositions on Transwell inserts. Inducibility and functionality of tightness were investigated by transendothelial electrical resistance (TEER) and by transport studies with transcellular marker diazepam, glycine antagonist Bu101 and paracellular marker APTS-dextran. Inducibility, expression and localization of tight junctional proteins were assessed by western blotting and immunofluorescence microscopy. Presence of factors derived from glioma cell line C6 resulted in increased TEER in both cell lines. Comparison to APTS-dextran data across Caco-2 layers emphasized that correlations between permeability of the paracellular marker and TEER are dependent on each investigated cell line and the corresponding growth medium. Presence and inducibility of tight junctional proteins ZO-1 and Occludin were proven for ECV304 layers. Cell line ECV304 seemed to be suitable for TEER dependent transport studies, whereas PBMEC/C1-2 showed higher potential for P-gP studies. PMID- 18399538 TI - Nestin expression defines both glial and neuronal progenitors in postnatal sympathetic ganglia. AB - Sympathetic ganglia are primarily composed of noradrenergic neurons and satellite glial cells. Although both cell types originate from neural crest cells, the identities of the progenitor populations at intermediate stages of the differentiation process remain to be established. Here we report on the identification in vivo of glial and neuronal progenitor cells in postnatal sympathetic ganglia, by using mouse superior cervical ganglia as a model system. There are significant levels of cellular proliferation in mouse superior cervical ganglia during the first 18 days after birth. A majority of the proliferating cells express both nestin and brain lipid-binding protein (BLBP). Bromodeoxyuridine (BrdU) fate-tracing experiments demonstrate that these nestin and BLBP double-positive cells represent a population of glial progenitors for sympathetic satellite cells. The glial differentiation process is characterized by a marked downregulation of nestin and upregulation of S100, with no significant changes in the levels of BLBP expression. We also identify a small number of proliferating cells that express nestin and tyrosine hydroxylase, a key enzyme of catecholamine biosynthesis that defines sympathetic noradrenergic neurons. Together, these results establish nestin as a common marker for sympathetic neuronal and glial progenitor cells and delineate the cellular basis for the generation and maturation of sympathetic satellite cells. PMID- 18399539 TI - Dynamic expression of the retinoic acid-synthesizing enzyme retinol dehydrogenase 10 (rdh10) in the developing mouse brain and sensory organs. AB - Organs develop through many tissue interactions during embryogenesis, involving numerous signaling cascades and gene products. One of these signaling molecules is retinoic acid (RA), an active vitamin A derivative, which in mammalian embryos is synthesized from maternal retinol by two oxidative reactions involving alcohol/retinol dehydrogenases (ADH/RDHs) and retinaldehyde dehydrogenases (RALDHs), respectively. The activity of RALDHs is known to be crucial for RA synthesis; however, recently a retinol dehydrogenase (RDH10) has been shown to represent a new limiting factor in this synthesis. We investigated the spatiotemporal distribution of Rdh10 gene transcripts by in situ hybridization and quantitative polymerase chain reaction (PCR) during development of the brain and sensory organs. Although Rdh10 relative mRNA levels decline throughout brain development, we show a strong and lasting expression in the meninges and choroid plexuses. Rdh10 expression is also specifically seen in the striatum, a known site of retinoid signaling. In the eye, regional expression is observed both in the prospective pigmented epithelium and neural retina. In the inner ear Rdh10 expression is specific to the endolymphatic system and later the stria vascularis, both organs being involved in endolymph homeostasis. Furthermore, in the peripheral olfactory system and the vibrissae follicles, expression is present from early stages in regions where sensory receptors appear and mesenchymal/epithelial interactions take place. The distribution of Rdh10 transcripts during brain and sensory organ development is consistent with a role of this enzyme in generating region-specific pools of retinaldehyde that will be used by the various RALDHs to refine the patterns of RA synthesis. PMID- 18399540 TI - Quantification of D1 and D5 dopamine receptor localization in layers I, III, and V of Macaca mulatta prefrontal cortical area 9: coexpression in dendritic spines and axon terminals. AB - D1 family receptors (D1R) in prefrontal cortex (PFC) are critical for normal cognition and are implicated in pathological states such as schizophrenia. The two D1R subtypes, D1 and D5, cannot be pharmacologically distinguished but have important functional differences. To understand their contributions to cortical function, we quantified their localization in the neuropil of primate PFC. We identified different patterns of distribution for the two receptors that showed variation across cortical laminae. Although D1 was enriched in spines and D5 in dendrites, there was considerable overlap in their distribution within neuronal compartments. To determine whether the D1 and D5 receptors are localized to separate populations of synapses, we employed double-labeling methods. We found the two receptors colocalized and quantified the overlap of their distribution in spines and axon terminals of prefrontal cortical area 9 in the Macaca mulatta monkey. The two receptors are found in partially overlapping populations, such that the D5 receptor is found in a subpopulation of those spines and terminals that contain D1. These results indicate that dopamine activation of the two D1R subtypes does not modulate disparate populations of synapses onto dendritic spines in prefrontal cortical area 9; rather, dopamine can activate D1 and D5 receptors on the same spines, plus an additional group of spines that contains only D1. The implications of these results for the dose-dependent relationship between D1R activation and PFC function are discussed. PMID- 18399542 TI - Tetrodotoxin-resistant voltage-gated sodium channels Na(v)1.8 and Na(v)1.9 are expressed in the retina. AB - Voltage-gated sodium channels (VGSCs) are one of the fundamental building blocks of electrically excitable cells in the nervous system. These channels are responsible for the generation of action potentials that are required for the communication of neuronal signals over long distances within a cell. VGSCs are encoded by a family of nine genes whose products have widely varying biophysical properties. In this study, we have detected the expression of two atypical VGSCs (Na(v)1.8 and Na(v)1.9) in the retina. Compared with more common VGSCs, Na(v)1.8 and Na(v)1.9 have unusual biophysical and pharmacological properties, including persistent sodium currents and resistance to the canonical sodium channel blocker tetrodotoxin (TTX). Our molecular biological and immunohistochemical data derived from mouse (Mus musculus) retina demonstrate expression of Na(v)1.8 by retinal amacrine and ganglion cells, whereas Na(v)1.9 is expressed by photoreceptors and Muller glia. The fact that these channels exist in the central nervous system (CNS) and exhibit robust TTX resistance requires a re-evaluation of prior physiological, pharmacological, and developmental data in the visual system, in which the diversity of VGSCs has been previously underestimated. PMID- 18399541 TI - Subcellular distribution of the Rho-GEF Lfc in primate prefrontal cortex: effect of neuronal activation. AB - The strength of synaptic connections in the brain varies with activity, and this plasticity depends on remodeling of the actin cytoskeleton in dendritic spines. Critical to this are the Rho family GTPases, whose activity is controlled by various modulatory proteins, including the Rho-GEF Lfc. In cultured neurons and nonneuronal cells, Lfc has been shown both to bind to microtubules and to regulate the actin cytoskeleton. Significantly, Lfc was found to be concentrated in the dendritic shafts of cultured hippocampal neurons under control conditions but then translocated into spines when neural activity was stimulated. In this study, we used immunohistochemistry and electron microscopy to examine activity dependent changes in the distribution of Lfc in the neuropil of monkey prefrontal cortex. We found that, although Lfc was concentrated in dendrites, it also had a complex distribution in the neuropil, including being present in spines, axons, terminals, and glial processes. Moreover, Lfc distribution varied in different layers of cortex. By using an in vitro slice preparation of monkey prefrontal cortex, we demonstrated an activity-dependent translocation of Lfc from dendritic shafts to spines. The results of this study support a role for Lfc in activity dependent spine plasticity and demonstrate the feasibility of studying activity dependent changes in protein localization in tissue slices. PMID- 18399543 TI - The random vibration effects on dissolution testing with USP apparatus 2. AB - Dissolution testing is of primary importance for optimization of drug formulation and quality control, but test results typically show large variability. Vibration is one of the factors that can increase variability of dissolution testing. In this study, a Distek USP Apparatus 2 was used to perform dissolution testing using disintegrating 10 mg prednisone tablets at 50 rpm in 500 mL of 37 degrees C degassed water medium. A controllable vertical random vibration was applied to the dissolution apparatus during the dissolution testing. Real-time vibration waveforms were recorded using accelerometers placed at various locations on the vessel plate and on the dissolution vessels. Preliminary results showed a strong correlation between induced vibration and dissolution results. The vibration measured on the vessel plate correlates well with that measured within nearby vessels. The observed dissolution profiles suggest that vibration affects the disintegration and dissolving processes by different mechanisms, leading to high or low results depending upon during which phase of the dissolution process the vibration occurs. This study also presents a method capable of measuring vibration in a meaningful manner and how to determine where best to measure it. PMID- 18399544 TI - Cryptochrome is present in the compound eyes and a subset of Drosophila's clock neurons. AB - Cryptochrome (CRY) is intimately associated with the circadian clock of many organisms. In the fruit fly Drosophila melanogaster, CRY seems to be involved in photoreception as well as in the core clockwork. In spite of the critical role of CRY for the clock of Drosophila, it was not quite clear whether CRY is expressed in every clock cell. With the help of a new antibody and a mutant that lacks CRY, we show here that CRY is expressed in specific subsets of Drosophila's pacemaker neurons and in the photoreceptor cells of the compound eyes. In the pacemaker neurons, CRY levels and kinetics under light-dark cycles are quite different from each other. High-amplitude oscillations are observed in only three groups of clock neurons, suggesting that these three groups are strongly receptive to light. The different CRY kinetics may account for phase differences in oscillations of the clock proteins observed in these three groups in earlier studies. The molecular clock of the neurons that contain lower CRY levels or are completely CRY negative can still be synchronized by light, probably via intercellular communication with the CRY-positive neurons as well as via external photoreceptors. PMID- 18399545 TI - Biomechanical evaluation of equine masticatory action: position and curvature of equine cheek teeth and age-related changes. AB - The equine cheek tooth battery is part of a very dynamic system. The aim of this study was to investigate whether the curvature and position of the teeth are also involved in such dynamical processes. The alveolar crest was labelled with a radiodense marker (48 cadaver heads, 15 skulls) and laterolateral radiographs were taken. Then a geometrical method was elaborated to determine a cheek tooth's curvature and its position by means of specific angles. This method respects the remarkable changes of the equine dentition throughout life by considering two items: (1) the alveolar crest was taken as a constant landmark, (2) the central axis of the curved dental crown was determined by calculation of a linear regression equation. This equation considered several geometrically determined points on the curved dental crown which had been marked in the radiographs. Our study yielded the following results: Mandibular cheek teeth became more curved with age, but their positions (represented by the so-called mesio-occlusal angle between tooth and alveolar crest) did not change significantly. In maxillary cheek teeth, however, the mesio-occlusal angle became larger with age (indication of change of dental position), while their curvature did not change. Even though changes of the dental position were not always statistically significant, they are discussed as being biologically/functionally relevant. The mandibular anticlinal tooth, i.e. the tooth positioned at a mesio-occlusal angle of about 90 degrees , was not in contact with the maxillary anticlinal tooth. Interestingly, the maxillary anticlinal tooth is known to cause most clinical dental problems. PMID- 18399546 TI - Distribution and corticosteroid regulation of glucocorticoid receptor in the brain of Xenopus laevis. AB - Glucocorticoids (GCs) play essential roles in physiology, development, and behavior that are mediated largely by the glucocorticoid receptor (GR). Although the GR has been intensively studied in mammals, very little is known about the GR in nonmammalian tetrapods. We analyzed the distribution and GC regulation of GR in the brain of the frog Xenopus laevis by immunohistochemistry. GR immunoreactive (GR-ir) cells were widely distributed, with the highest densities in the medial pallium (mp; homolog of the mammalian hippocampus), accumbens, anterior preoptic area (POA; homolog of the mammalian paraventricular nucleus), Purkinje cell layer of the cerebellum, and rostral anterior pituitary gland (location of corticotropes). Lower but distinct GR-ir was observed in the internal granule cell layer of the olfactory bulbs, dorsal and lateral pallium, striatum, various subfields of the amygdala, bed nucleus of the stria terminalis (BNST), optic tectum, various tegmental nuclei, locus coeruleus, raphe nuclei, reticular nuclei, and the nuclei of the trigeminal motor nerves. Treatment with corticosterone (CORT) for 4 days significantly decreased GR-ir in the POA, mp, medial amygdala (MeA), BNST, and rostral pars distalis. Treatment with the corticosteroid synthesis inhibitor metyrapone (MTP) also significantly reduced GR ir in the POA, mp, MeA and BNST, but not in the rostral pars distalis. Replacement with a low dose of CORT in MTP-treated animals reversed these effects in brain. Thus, chronic increase or decrease in circulating corticosteroids reduces GR-ir in regions of the frog brain. Our results show that the central distribution of GR-ir and regulation by corticosteroids are highly conserved among vertebrates. PMID- 18399548 TI - N,N'-Bis(5-aminosalicyl)-L-cystine is a potential colon-specific 5-aminosalicylic acid prodrug with dual therapeutic effects in experimental colitis. AB - To evaluate N,N'-bis(5-aminosalicyl)-L-cystine (5-ASA-Cys) as a potential colon specific 5-aminosalicylic acid prodrug with dual therapeutic effects in experimental colitis, the pharmacokinetics and therapeutic activity were investigated after oral administration of 5-ASA-Cys and amelioration of experimental colitis was compared after rectal administration of 5-aminosalicylic acid (5-ASA) and/or cysteine. In addition, the gluthathione (GSH) level in the inflamed colonic tissue was examined after administration of cysteine or 5-ASA Cys. Oral administration of 5-ASA-Cys delivered much greater amount of 5-ASA to the large intestine and excreted lower amount of 5-ASA via urine than that of free 5-ASA. Oral administration of 5-ASA-Cys ameliorated experimental colitis of rats induced by TNBS, which was more effective than that of sulfasalazine. Although cysteine administered rectally was not significantly effective, intracolonic treatment with both 5-ASA and cysteine showed a synergic effect in alleviating the rat colitis. Furthermore, not only 5-ASA-Cys administered orally but also cysteine administered rectally increased the glutathione level in the inflamed colonic tissue. Taken together, these results suggest that 5-ASA-Cys is a potential colon specific 5-ASA prodrug with dual therapeutic effects on experimental colitis and cysteine modulation of the glutathione level may be relevant to the dual effects of the prodrug. PMID- 18399549 TI - Effect of gonadal hormones on the cross-sectional area of pubococcygeus muscle fibers in male rat. AB - Effects of gonadal hormones on dimorphic striated muscles such as the bulbocavernosus/levator ani complex related to male penile erection have been widely studied. However, the action of these hormones on pelvic nondimorphic muscles is not known. In the present study, the sensitivity of the male rat pubococcygeus muscle (Pcm) to gonadal hormones was studied measuring the cross sectional area (CSA) of its fibers. For this, two experiments were done: in the first, the effect of castration, and in the second the effect of gonadal hormone administration was analyzed. We found that castration after 6 weeks significantly reduced the average CSA of the fibers of this muscle and that castration after 2 or 6 weeks reduced the percentage of fibers with higher CSAs, but only castration after 6 weeks increased the percentage of fibers with the lowest CSA. In comparison with castrated animals implanted with an empty Silastic capsule, Silastic implants of testosterone propionate or dihydrotestosterone significantly increased the average CSA of Pcm fibers, and the treatment with testosterone propionate, estradiol benzoate, or dihydrotestosterone decreased the percentage of fibers with low CSAs and increased the percentage with larger CSAs. Our results could be considered for therapy in patients with damage of the Pcm, and suffering urinary incontinence or ejaculatory dysfunctions. PMID- 18399550 TI - ING4 induces cell growth inhibition in human lung adenocarcinoma A549 cells by means of Wnt-1/beta-catenin signaling pathway. AB - ING4, as a novel candidate tumor suppressor gene, has been implicated in several human malignances by tumor growth inhibition and apoptosis enhancement. The mechanism of ING4 remains largely unknown. The purpose of this study was to investigate the inhibitory tumor growth effects of ING4 on lung adenocarcinoma, and its mechanism, by ING4 cDNA transduction into A549 cells. Furthermore, the expression level of ING4 in lung adenocarcinoma tissues was examined. The expression of ING4 was markedly reduced in human lung adenocarcinoma tissues. Overexpression of ING4 can induce growth inhibition in A549 cells both in vitro and in vivo, and also induce up-regulation of p27, down-regulation of cyclinD1, SKP2, and Cox2, and inactivation of the Wnt-1/beta-catenin pathway. Moreover, overexpression of ING4 can enhance the sensitivity of A549 cells to radiotherapy and chemotherapy. Thus, ING4 may play an inhibitory role on A549 cell proliferation and tumor growth in lung adenocarcinoma by up-regulation or down regulation of cell proliferation-regulating proteins such as p27, cyclinD1, SKP2, and Cox2 by means of inactivation of Wnt-1/beta-catenin signaling. PMID- 18399551 TI - A concise synthesis of (-)-oseltamivir. PMID- 18399552 TI - Synthesis of (-)-octalactin a by a strategic vanadium-catalyzed oxidative kinetic resolution. PMID- 18399553 TI - Heterotrimetallic salts: synthesis, structures, and superbase reactivity of crystalline tert-butoxides [Li4Na2K2(OtBu)8(mu-L)]n. PMID- 18399554 TI - Protein-cross-linked polymeric materials through site-selective bioconjugation. PMID- 18399555 TI - Total synthesis of (+)-oocydin A: application of the Suzuki-Miyaura cross coupling of 1,1-dichloro-1-alkenes with 9-alkyl 9-BBN. PMID- 18399556 TI - Diphenylprolinol silyl ether as a catalyst in an enantioselective, catalytic, formal aza [3+3] cycloaddition reaction for the formation of enantioenriched piperidines. PMID- 18399557 TI - Development of selective RabGGTase inhibitors and crystal structure of a RabGGTase-inhibitor complex. PMID- 18399558 TI - Fluorescence-quenching-based enzyme-activity assay by using photon upconversion. PMID- 18399559 TI - A caged retinoic acid for one- and two-photon excitation in zebrafish embryos. PMID- 18399561 TI - A highly efficient and selective Au(I)-catalyzed tandem synthesis of diversely substituted pyrrolo[1,2-a]quinolines in aqueous media. PMID- 18399562 TI - Simultaneous determination of the conformation and relative configuration of archazolide a by using nuclear overhauser effects, J couplings, and residual dipolar couplings. PMID- 18399563 TI - Formulation strategies to minimize oxidative damage in lyophilized lipid/DNA complexes during storage. AB - It has been shown that degradation of lipid/DNA complexes (lipoplexes) continues in the dried state during storage. The goal of this study was to evaluate the ability of various strategies to minimize the formation of reactive oxygen species (ROS) in lyophilized lipoplexes during storage, including metal removal from reagents, air displacement, and fortification with chelator/antioxidant agents. Formulations containing individual chelator (DTPA) and antioxidants (L methionine or alpha-tocopherol), or in combination, were subjected to lyophilization. Accelerated storage conditions were investigated and physico chemical characteristics and biological activity of samples were monitored at different time intervals. Generation of ROS during storage was determined by adding proxyl fluorescamine to the formulations prior to freeze-drying. Lipid peroxidation was assessed by monitoring the formation of thiobarbituric reactive substances (TBARS) and lipid hydroperoxides. We also assessed the effect of increased moisture content on the chemical and biological stability of lipoplexes containing additives. Our results show that both ROS and TBARS are generated in lyophilized cakes during storage, and that agents such as DTPA or alpha tocopherol are efficient in protecting lipid/DNA complexes against oxidative damage in the dried state. Our experiments also indicate that higher residual moisture has a deleterious effect on the stability of lipid/DNA complexes during storage. PMID- 18399564 TI - Pharmacokinetics and tissue distribution of 8,9-epoxy brevifolin in rats, a hepatoprotective constituent isolated from Phyllanthus simplex Retz by liquid chromatography coupled with mass spectrometry method. AB - 8,9-Epoxy brevifolin (EBF) is a novel compound isolated from Phyllanthus simplex Retz (P. simplex) and has been demonstrated to possess a hepatoprotective effect. The purposes of the present study were to examine the in vivo pharmacokinetics and tissue distribution of EBF in rats using a liquid chromatography coupled with mass spectrometry quantitative detection method (LC-MS/MS), with luteolin-7-O glucoside being employed as an internal standard (IS). The method was validated within the concentration range 20-15 000 ng/ml, and the calibration curves were linear with correlation coefficients of >0.999. The limit of quantitation (LOQ) for EBF was 20 ng/ml. The intra-assay accuracy and precision ranged from 98.7% to 100.2% and 2.19% to 6.25%, respectively, while the inter-assay accuracy and precision ranged from 97.5% to 100.3% and 3.35% to 7.28%, respectively. The method was further applied to assess the pharmacokinetics and oral bioavailability of EBF after intravenous and oral administration in rats. The oral bioavailability of EBF was 12.46 +/- 2.31%. In the tissue distribution assay, its concentration was higher in the heart (13.2 +/- 0.24 microg/g) and liver (14.5 +/- 0.19 microg/g) than in other tissues. PMID- 18399565 TI - The changing face of orthopaedic trauma: locked plating and minimally invasive techniques. AB - Rapid advances in the field of orthopaedic trauma have improved treatment options while keeping pace with the changing characteristics of the trauma population. The availability of locking implants has changed the approach to treating fractures in older patients with osteoporotic bones as well as in those with comminuted and complex injuries. Minimally invasive approaches have allowed the preservation and protection of soft tissues while allowing adequate reduction and fixation of fractures. This biologically friendly approach coupled with newer implants and instruments will improve early and long-term outcomes in trauma care. PMID- 18399566 TI - The changing face of orthopaedic trauma: fragility and periprosthetic fractures. AB - Because fragility fractures and periprosthetic fractures represent an increasingly common source of morbidity for the elderly population, all orthopaedic surgeons should be prepared to care for such injuries in their practice. Fractures in elderly patients appear to heal more slowly because of decreased numbers and responsiveness of osteoprogenitor stem cells, and stable fixation is more difficult to achieve. It is important to review the mechanical and biologic considerations relevant to the treatment of these challenging injuries. PMID- 18399567 TI - Technical tips in fracture care: fractures of the hip. AB - Hip fracture is an increasingly common and clinically significant injury with substantial economic impact. Associated risk factors are age, gender, race, bone density, activity level, and medical disorders. Prevention efforts include treatment of osteoporosis and programs to reduce the risks of a fall. Nondisplaced or impacted fractures of the femoral neck can be treated with screw fixation. Displaced femoral neck fractures in younger, more active patients may be treated with reduction and fixation. In physiologically older patients, joint arthroplasty is indicated for displaced fractures. In patients with systemic arthritis or preexisting hip disease, total hip arthroplasty may be an appropriate treatment choice. Intertrochanteric fractures are treated with reduction and fixation using either a sliding hip screw and side plate or intramedullary nail with cephalic interlock. Key technical points for successful outcomes include proper patient positioning, using a correct starting point for the nail, achieving acceptable reduction before fixation, and the use of various reduction techniques and aids. PMID- 18399568 TI - Locking plates: tips and tricks. AB - Locking plates are fracture fixation devices that allow the insertion of fixed angle/angular-stable screws or pegs and do not require friction between the plate and bone. The clinical care impetus for the development of these plates has been a combination of factors, including the increasing survival of patients with high energy injuries, aging Western European and North American populations with an increasing rate of fragility fractures, and dissatisfaction of patients and surgeons with the outcomes of treatment of specific periarticular fractures. Nonclinical factors likely include a push by industry for new technology and new markets as well as the general interest of the public in "minimally invasive" surgery. PMID- 18399569 TI - External fixation: how to make it work. AB - The external fixator has been in use for more than a century. Wutzer (1789-1863) used pins and an interconnecting rod-and-clamp system. Parkhill (1897) and Lambotte (1900) used devices that were unilateral with four pins and a bar-clamp system. By 1960, Vidal and Hoffmann had popularized the use of an external fixator to treat open fractures and infected pseudarthroses. The complications associated with the use of external fixation in the late 20th century were predominantly caused by a lack of understanding of the principles of application, the principles of fracture healing with external fixation, and old technology. Its use was reserved for the most severe injuries and for cases complicated by infection. Thus, pin problems, nonunions, and malunions were common. Better technology and understanding have since allowed for greater versatility and better outcomes. Simultaneous with developments in the Western world, Ilizarov developed the principles of external fixation with use of ring and wire fixation. It was not until the late 1980s and early 1990s, when more interaction and exchange between the West and East (Russia) became possible, and with the help of Italians who embraced the philosophy of external fixation, that the use of external fixation was proven to be successful. Several variations of external fixation have been developed, and its use is now widespread. However, in the United States, all but a minority of surgeons still have substantial apprehension about the use of external fixation. PMID- 18399570 TI - Management of open fractures and subsequent complications. AB - Early, systemic, wide-spectrum antibiotic therapy is necessary for the treatment of open fractures. The bead pouch technique delivers antibiotics locally and prevents secondary wound contamination. The open fracture wound should be thoroughly debrided. To avoid the complication of gas gangrene, the wound should not be closed. Extensive soft-tissue damage may necessitate the use of local or free flaps. Techniques of fracture stabilization depend on the anatomic location of the fracture and the characteristics of the injury. Early bone grafting and supplemental procedures may be needed to achieve healing. Management of the infected open fracture is based on radical debridement, skeletal stabilization, microbial-specific antibiotics, soft-tissue coverage, and reconstruction of bone defects. PMID- 18399571 TI - Orthopaedic war injuries: from combat casualty care to definitive treatment: a current review of clinical advances, basic science, and research opportunities. AB - Musculoskeletal war wounds often involve massive injury to bone and soft tissue that differ markedly in character and extent compared with most injuries seen in civilian practice. These complex injuries have challenged orthopaedic surgeons to the limits of their treatment abilities on the battlefield, during medical evacuation, and in subsequent definitive or reconstructive treatment. Newer methodologies are being used in the treatment of these wounds to prevent so called second hit complications, decrease complications associated with prolonged medical evacuation, reduce the incidence of infection, and restore optimal function. Basic science advances hold the promise of providing foundations for future treatment options that may improve both bone and soft-tissue healing. Research on the treatment of these often devastating wounds also will have broad applicability to trauma resulting from acts of terrorism or from natural disasters. PMID- 18399572 TI - From Iraq back to Iraq: modern combat orthopaedic care. AB - War wounds are often large and complex, with high degrees of contamination and tissue loss differing significantly from typical civilian injuries. Infection has been a common complication driving the tenets of care, even in the antibiotic age. Fractures were historically treated with casting or traction because of the risk of infection with internal fixation. However, current civilian fracture care has evolved significantly with extensive use of internal and external fixation with early mobilization and other adjuncts to restore function earlier and more completely. Whether the application of modern techniques and implants can better restore function in patients with these severe injuries is currently being evaluated. PMID- 18399573 TI - What went wrong and what to do about it: pitfalls in the treatment of shoulder impingement. AB - Shoulder impingement is a commonly treated disorder; however, the absence of a clear understanding of pathophysiology and causation can create variability in treatment. Pitfalls that can impede optimal patient outcomes can occur in the areas of diagnosis, treatment, and rehabilitation. Diagnostic pitfalls include failing to identify some of the multiple extrinsic diagnoses as well as the intrinsic subacromial pathology in patients with symptoms of impingement. Surgical pitfalls include not addressing the entire pathology and doing an incomplete or excessive decompression. Rehabilitation pitfalls include prescribing the wrong exercises at the wrong time and omitting steps in the rehabilitation process. Every patient who presents for primary treatment of impingement or secondary treatment following failed treatment should have a thorough evaluation of all intrinsic and extrinsic factors that can contribute to the symptoms of shoulder impingement. PMID- 18399574 TI - Complications in arthroscopic anterior shoulder stabilization: pearls and pitfalls. AB - Arthroscopic treatment of anterior glenohumeral instability has become increasingly common. As longer-term follow-up studies become available, certain trends dictating the success or failure of arthroscopic stabilization are becoming more evident. Bone defects are important predictors of clinical failure, and the recognition of bone loss and other pathoanatomic variables can help determine which patients will benefit from arthroscopic stabilization for anterior glenohumeral instability. Arthroscopic techniques for anterior shoulder instability must mirror the focus of open methods on retensioning the inferior glenohumeral ligament and restoring the anatomy of the anterior capsulolabral complex. Arthroscopic stabilization for anterior glenohumeral instability has achieved results comparable to those of open stabilization methods in properly selected patients. Advantages of arthroscopic treatment of shoulder instability include lower morbidity, decreased pain, the ability to treat other pathologies, and improved cosmesis. As arthroscopic treatment of recurrent shoulder instability becomes more commonplace, it is crucial to review the factors that influence the success or failure of arthroscopic instability procedures and to understand the guidelines for patient selection, surgical pearls and pitfalls, and adjunctive technical details designed to optimize results. PMID- 18399575 TI - Arthroscopic anterior shoulder instability repair: techniques, pearls, pitfalls, and complications. AB - Initial attempts to replicate the success of open anterior shoulder instability procedures using arthroscopic procedures were associated with unacceptably high failure rates. The resultant focus on identifying clear surgical indications and improving both arthroscopic technique and instrumentation has culminated in arthroscopic success rates approaching those of established open procedures. Current experience shows that with careful patient selection, strict adherence to proper surgical technique, and the avoidance of common surgical errors, excellent clinical results can be achieved with arthroscopic instability repair. PMID- 18399576 TI - Posterior and multidirectional instability of the shoulder: challenges associated with diagnosis and management. AB - The diagnosis and treatment of posterior and multidirectional instability of the shoulder remain challenging because of complexities in the classification and etiology of the condition and the physical examination of a patient with suspected posterior and multidirectional instability. With an improved understanding of pertinent clinical symptoms and physical examination findings, a successful strategy for nonsurgical and surgical management can be developed. It is essential to understand the biomechanics of repair procedures, relevant pathoanatomy, and techniques of surgical management to optimize surgical success. Arthroscopic techniques have allowed improved diagnosis and treatment of this condition. PMID- 18399577 TI - Massive irreparable tendon tears of the rotator cuff: salvage options. AB - The management of massive, irreparable rotator cuff tears is challenging. Arthroscopic debridement has produced reasonable short-term outcomes in patients who experience good relief from pain and improved range of motion after a subacromial injection with local anesthetic. Arthroscopic debridement with partial repair also has resulted in good outcomes, especially in patients with documented suprascapular nerve traction neurapraxia. Tendon transfer can offer a long-lasting solution in a patient with a weak shoulder who still has the ability to raise the shoulder past the horizontal position. Shoulder replacement with the use of a reverse prosthesis has emerged as a viable option in patients with pseudoparalysis with or without osteoarthritis of the glenohumeral joint. PMID- 18399578 TI - The reverse total shoulder arthroplasty. AB - A reverse total shoulder arthroplasty is a prosthesis that should be used in patients who have specific contraindications to the more conventional total shoulder prosthetic replacements. The patient and surgeon should understand that this reconstruction is technically more difficult and is associated with more complications than conventional shoulder reconstructions. The reverse total shoulder arthroplasty has been used in Europe more than in North America, and the experience in Europe is guiding its use in North America. An understanding of the mechanics of the reverse total shoulder arthroplasty and the technical details of its implantation will help in understanding its role in shoulder reconstruction. PMID- 18399579 TI - Median and radial nerve compression about the elbow. AB - It is important for physicians who treat upper extremity disorders to understand motor palsy or pain syndromes caused by compression of the median and radial nerves about the elbow and forearm. Patients with anterior interosseous nerve syndrome may report hand weakness, whereas those with pronator syndrome may present with pain and paresthesia that can be confused with carpal tunnel syndrome. Patients with posterior interosseous nerve syndrome report hand weakness, whereas those with radial tunnel syndrome report pain in the lateral elbow and forearm, which may be confused with lateral epicondylitis. Because each syndrome has overlapping symptoms, serial examinations are needed to determine the correct diagnosis. PMID- 18399580 TI - Compression of the ulnar nerve at the elbow: cubital tunnel syndrome. AB - Although cubital tunnel syndrome has been described as the most common entrapment of the ulnar nerve, there is still considerable difficulty identifying the exact location of the pathologic compression of the nerve and deciding on the correct surgical or nonsurgical treatment. The most commonly recommended surgical techniques include simple (in situ) decompression, decompression with medial epicondylectomy, anterior subcutaneous transposition, and anterior submuscular transposition of the ulnar nerve at the elbow. It is important to understand the pitfalls and possible complications of these commonly used treatments. PMID- 18399581 TI - Controversies in carpal tunnel syndrome. AB - Compression of the median nerve at the wrist (carpal tunnel syndrome) is the most common compression neuropathy of the upper extremity. Surgical decompression has become one of the most frequently performed peripheral nerve procedures. Despite the popularity of the procedure, and the volume of clinical and basic science information available, differences of opinion exist as to diagnosis and management. This review discusses several of these controversies and presents both recent and historical data that have led to some of these disagreements. PMID- 18399582 TI - Minimally invasive total hip arthroplasty: an overview of the results. AB - Small-incision total hip arthroplasty (THA) has been shown to be safe and effective in achieving early postoperative improvements in pain and function. The comparative published reports of the two-incision, anterior, and mini-posterior techniques have defined indications for small-incision THAs. The mini-posterior approach appears to be better than the traditional posterior approach for THA in terms of early patient function and acceptance. There may be little difference among the mini-incision techniques when preoperative patient education and postoperative rehabilitation are equivalent. Correct component positioning has been consistently achieved with small-incision procedures, and short-term results are the same as those of traditional THA. The mini-posterior approach also has been shown to have psychological advantages because it allows patients to be more confident about their outcomes. New anesthesia and pain management techniques have also improved early functional results. With time and technical advances such as computer navigation, the use of minimally invasive THA will become more prevalent. PMID- 18399583 TI - Muscle damage in minimally invasive total hip arthroplasty: MRI evidence that it is not significant. AB - The prevalence of damage to the musculature surrounding the hip joint was assessed in a random selection of patients who had a unilateral primary total hip replacement with either the two-incision minimally invasive technique, the standard posterolateral approach, or the direct lateral approach. The musculature of the operated hip was compared with that of the normal contralateral side using MRI with a special metal subtraction protocol that allowed visualization of the muscles and tendons while minimizing metal artifacts. All patients had undergone total hip arthroplasty at least 18 months before the investigation. The data show that the standard posterolateral and direct lateral approaches are associated with an increased incidence of postoperative alterations in the hip muscles after total hip arthroplasty, when compared with two-incision minimally invasive hip replacement. The results from this study suggest that the two-incision technique for total hip replacement may have muscle-sparing advantages over other standard approaches. PMID- 18399584 TI - Muscle damage during minimally invasive total hip arthroplasty: cadaver-based evidence that it is significant. AB - Minimally invasive total hip arthroplasty has generated substantial interest in both patients and surgeons. The concept that smaller incisions and less extensive surgical dissection should lead to less pain and a quicker recovery is inherently appealing. Advocates of minimally invasive total hip arthroplasty have suggested that some minimally invasive total hip approaches can be done without cutting any muscle or tendon. This contention has been carefully examined through a series of comparative cadaver studies, and the authors have determined that it is not possible to routinely perform minimally invasive total hip arthroplasty without causing some measurable degree of muscle damage. Anatomic studies showed that the two-incision approach with fluoroscopy, the posterior mini-incision approach, and the mini-incision Smith-Petersen approach all were associated with measurable muscle damage. The clinical importance of this muscle damage cannot be answered by these cadaver studies. PMID- 18399585 TI - Surgical nuances to minimize muscle damage during the direct lateral approach in minimally invasive hip replacement. AB - Compared with total hip replacements performed using a standard incision direct lateral approach, a minimally invasive direct lateral approach can produce less pain and blood loss without increasing the rate of complications. To minimize the potential for limping after detaching the anterior gluteus medius and the gluteus minimus tendons, special handling of the abductors (which is more difficult because of the limited exposure afforded by the smaller incision) is required. Using special retractors, minimizing overzealous retraction, and achieving strong repair has resulted in no increase in the incidence of limping. PMID- 18399586 TI - Surgical nuances to minimize muscle damage during the anterolateral intermuscular approach in minimally invasive hip replacement. AB - Muscle damage that can occur during minimally invasive total hip replacement is an important concern. Minimizing this iatrogenic injury can help achieve the goals of the minimally invasive approach: decreased postoperative pain, decreased blood loss, and faster rehabilitation. Knowledge of particular aspects of minimally invasive anterolateral total hip arthroplasty is important, with focus on the nuances that aid in reducing muscle injury. PMID- 18399587 TI - Functional recovery of muscles after minimally invasive total hip arthroplasty. AB - Whether mini-incision total hip arthroplasty is associated with accelerated postoperative recovery is a subject of considerable controversy. A study was conducted to compare objective outcomes using gait analysis as a measure for recovery of function in patients treated with three different minimally invasive surgical approaches and the traditional posterior approach. Sixty-nine patients underwent instrumented gait analysis at self-selected and fast velocities preoperatively and at 6 weeks and 3 months postoperatively. Four surgical groups were studied-30 treated with posterior mini-incisions, 11 anterolateral, 10 anterior Judet, and 18 traditional posterior long incisions. Overall, gait velocity increased slightly at 6 weeks and significantly at 3 months. However, there were no significant differences between groups for velocity, cadence, stride length, single-limb support time, or double-limb support time at 6 weeks or 3 months postoperatively. These data indicate that patients undergoing total hip arthroplasty with any of these surgical approaches recover muscle function, as measured by gait analysis, to preoperative levels within 6 weeks postoperatively. No advantage was shown with the use of any of the three different small-incision approaches. This finding suggests that the amount of muscle, or the particular muscle cut, does not have a significant effect on the recovery of postoperative gait function. PMID- 18399588 TI - Hot topics and controversies in arthroplasty: cementless femoral fixation in elderly patients. AB - Cementless femoral fixation has been established as the gold standard for hip arthroplasty in young patients because of its exceptional longevity. Because older Americans are living longer and staying active, cementless femoral fixation for hip arthroplasty should be considered in all patients who have good bone quality. Numerous studies have shown excellent results using cementless fixation for hip arthroplasty in elderly patients. Histologic analysis, radiographic review, and dual-energy x-ray absorptiometry have shown solid osseointegration for biologic fixation and minimal bone loss. Cementless fixation provides superb functional outcomes with results comparable to those achieved using cemented fixation for hip arthroplasty. Additional advantages of cementless femoral fixation include shorter surgical times and substantial savings in health care costs. PMID- 18399589 TI - Cement fixation of the femoral component in older patients. AB - Polymethylmethacrylate cement fixation of the femoral component in total hip arthroplasty is still a reasonable option in a select group of patients. Cement fixation is indicated in sedentary patients older than 75 years, certain older patients with rheumatoid arthritis, and patients with femoral neck fractures or conversion surgery. Hips with severe osteopenia or a "stove-pipe" femur (Dorr type C) may also be considered for cement fixation. The cemented femoral component should be fabricated of a cobalt-chromium alloy with a modern geometry and offset. Third-generation cementing techniques should be used to obtain a grade A or B cement mantle. Antibiotic cement may be used in patients at higher risk for infection. The 10-year results of cemented femoral components in these patient populations are excellent. PMID- 18399590 TI - Hip resurfacing arthroplasty: time to consider it again? No. AB - Total hip arthroplasty (THA) is one of the most successful operations in orthopaedics. A new procedure designed to replace THA, such as hip resurfacing arthroplasty, even for select indications, must offer a definite improvement over the well-established gold standard of treatment. Hip resurfacing does not currently meet this standard. THA is a proven and durable procedure with excellent results and superior short-term implant survival compared with hip resurfacing arthroplasty. Patients treated with hip resurfacing arthroplasty incur unique risks associated with implant malpositioning resulting from the surgeons' steep learning curve, the complex instrumentation involved, and the technical difficulty of the procedure, as well as a risk of femoral neck fracture. Hip resurfacing has limited ability to appropriately restore hip biomechanics and limb length, and concerns for the effects of metal ion and potential revision challenges remain. To date, there is a lack of literature supporting the claim of superior functional outcomes in hip resurfacing compared with THA. Reconsideration of hip resurfacing arthroplasty is unwarranted until appropriate comparative studies can demonstrate a clear benefit to patients. PMID- 18399591 TI - Digital templating in total hip arthroplasty. AB - Preoperative templating for total hip arthroplasty requires precision and accuracy. The current method of acetate overlays is subject to errors in magnification. Digital radiography permits the use of software programs that calculate the x-ray magnification with precision and then adjust the templates to match the exact magnification. A reproducible set of steps is described that allows a surgeon to preoperatively template in the digital environment. PMID- 18399592 TI - Extensor mechanism complications after total knee arthroplasty. AB - Extensor mechanism complications after total knee arthroplasty are relatively uncommon but potentially devastating. The etiology of these complications, which includes patellofemoral instability, periprosthetic patellar fracture, and disruptions of the quadriceps tendon and patellar ligament, has become better defined in recent years, with a subsequent decrease in the incidence, primarily resulting from changes in surgical technique and component design. In addition to addressing the patient's specific failure mechanism, the treatment of extensor mechanism complications after total knee arthroplasty may include nonsurgical management, primary repair, or reconstruction with autogenous, allogeneic, or synthetic tissue. Prevention of these complications, the foremost goal, is achieved through identification of patient and procedure risk factors, meticulous surgical technique, including vigilance during exposure and retractor placement, and a high index of suspicion both during and after the procedure. PMID- 18399593 TI - Instability after total knee arthroplasty. AB - Instability after total knee arthroplasty is reported to result in implant failure, and substantial instability often requires revision surgery. Successful outcomes can be achieved after revision total knee arthroplasty, particularly if the etiology of the instability is identified before the revision procedure. After careful clinical and radiologic analysis, instability can be classified as extension instability, flexion instability, or genu recurvatum. It is important to understand the causes and recommended treatments of each type of instability. PMID- 18399594 TI - Infected total knee arthroplasty: diagnosis and treatment. AB - Infection following total knee arthroplasty is a challenging complication for both the patient and the surgeon. Precautions to prevent infection include the use of prophylactic antibiotics, minimized surgical time, and meticulous surgical technique. A patient's risk factors should be thoroughly assessed and medical comorbidities should be treated before surgery. When infection is suspected, prompt evaluation of the patient is necessary. The management of a TKA infection is dictated by the duration of symptoms, suspicion for infection, time since index surgery, the patient's willingness and medical capability for undergoing multiple surgeries, the bacteria's sensitivity to antibiotics, and the surgeon's ability to perform complex surgery. Acute infections are treated with aggressive debridement and parenteral antibiotics. In appropriate circumstances, chronic infections are best treated with two-stage reimplantation, which can reliably eradicate infection, relieve pain, and restore good function. PMID- 18399595 TI - Infection in primary total knee arthroplasty: contributing factors. AB - Infection after a total knee arthroplasty is an infrequent but serious complication that can have devastating consequences. Infection carries a risk of significant morbidity, and the cost of treatment can be a substantial burden to the health care system. Eradication of infection can be very difficult. Prevention of infection remains the ultimate goal. Identification of host risk factors, careful patient selection, and optimization of the wound environment and the operating room remain some of the core fundamental steps that help minimize the overall incidence of infection. Although the exact role of each of these risk factors in a clinical setting can be debatable, a multidisciplinary approach incorporating all known and established methods of infection control can help to minimize the incidence of infection following total knee arthroplasty. PMID- 18399596 TI - Diagnosis and treatment of the infected primary total knee arthroplasty. AB - A diagnosis of infection in the painful primary total knee replacement is not always a straightforward endeavor. No single, fail-proof diagnostic study for infection exists. Often multiple diagnostic studies that include imaging, blood work, and joint aspiration as well as history and physical examination need to be considered. Infection may not always be determined before surgery, in which case intraoperative frozen sections can help to confirm infection or refute a negative workup. Treatment options vary, depending on the timing in the infection process and the source of the infection and may consist of simpler treatment courses, such as irrigation, debridement, and polyethylene exchange, to more complex treatment courses, such as two-stage revision with an antibiotic spacer to fusion or amputation. The orthopaedic surgeon uses an essential armamentarium of diagnostic and treatment options to determine the presence of infection and tailor the individual treatment of each patient. PMID- 18399597 TI - Revision total knee arthroplasty: planning, management, and controversies. AB - The number of total knee arthroplasties (TKAs) performed annually in the United States is increasing exponentially. Even with modest annual revision rates, the number of patients requiring revision knee surgery will increase in a similar fashion. It is therefore important to have a systematic approach dealing with a patient presenting with a painful TKA. The treating physician must be able to recognize a variety of failure patterns and treat them accordingly. Surgical exploration in the absence of a definable cause is rarely successful and should be avoided. The ability to plan and execute a complex revision TKA is a challenging and rewarding aspect of adult reconstructive surgery. Commonly encountered mechanisms of failure are identified and a stepwise approach to the surgical management of these conditions is presented. PMID- 18399598 TI - Controversies and techniques in the surgical management of patellofemoral arthritis. AB - Historically, the patellofemoral articulation has been a topic of less interest among orthopaedists and has been subject to fewer studies when compared with other major joints in the body. Patellofemoral arthritis is a common and debilitating condition, and greater awareness of this has led to a new interest and recent increase in the number of clinical investigations pertaining to this condition. It is hoped that an overview of patellofemoral kinematics, forces, and contact patterns will help in understanding the progression of patellofemoral arthritis. Furthermore, this understanding will ultimately allow the surgeon to apply these basic principles to more effective nonsurgical and surgical treatment options. Treatment methods for patellofemoral arthritis include both conservative as well as surgical interventions. Specifically, these treatments range from stretching and water exercises to patellar realignment procedures and the recently developed procedure of custom patellofemoral arthroplasty. In addition, many new and innovative treatments are on the horizon. This renewed interest in the patellofemoral articulation bodes well for patients who suffer from this condition. PMID- 18399599 TI - Total ankle arthroplasty. AB - Recent investigations support the belief that ankle replacement represents an attractive surgical alternative to arthrodesis for patients with advanced ankle arthritis. Although longer follow-up is necessary for total ankle arthroplasty (TAA) to displace arthrodesis as the surgical "gold standard," intermediate-term results are encouraging. Indications for TAA include primarily posttraumatic and inflammatory arthritis. Contraindications to TAA include unresectable osteonecrotic bone, peripheral vascular disease, neuropathy, active and/or recent ankle infection, nonreconstructible ankle ligaments, loss of lower leg muscular control, and severe osteopenia or osteoporosis. Young, active, high-demand patients with ankle arthritis may be better candidates for arthrodesis than for TAA. Rigorous patient selection is essential in the success of TAA, more than in other joint arthroplasty procedures. Total ankle prosthetic designs (Agility, Scandinavian Total Ankle Replacement, Hintegra, Salto, and Buechel-Pappas) with a minimum of published intermediate follow-up results, and several other innovative and biomechanically supported designs (the Mobility Total Ankle System, BOX, INBONE, and Salto-Talaris) are reviewed to demonstrate the recent evolution of TAA. Some TAA designs feature a nonconstrained polyethylene meniscus (mobile bearing) that articulates between the porous-coated tibial and talar components. The concern for edge loading (when the polyethylene component comes in contact with a metal edge) has been addressed in more recent designs by reducing the superior polyethylene surface area, expanding the tibial component surface, and even offering a convex tibial component. More practical, effective, and safer instrumentation for implantation has also been developed and has been essential to the success of TAA. However, complications with TAA (such as inadequate wound healing and malleolar fractures) are more frequent when compared with total hip and knee arthroplasty, irrespective of the surgeon's training method. As an individual surgeon gains more experience, the chances of a favorable outcome are increased. PMID- 18399600 TI - Surgical correction of moderate and severe hallux valgus: proximal metatarsal osteotomy with distal soft-tissue correction and arthrodesis of the metatarsophalangeal joint. AB - Hallux valgus correction by distal soft-tissue release and proximal metatarsal osteotomy is the procedure of choice for most patients with moderate and severe hallux valgus deformity. Complications can be avoided by selecting a procedure that provides adequate correction of the intermetatarsal angle and ensuring proper balancing of the metatarsophalangeal joint though lateral soft-tissue releases and medial joint plication. Arthrodesis should be considered when revision of failed surgery is planned, degenerative joint disease is present, and where the likelihood of failure of a bunion procedure is high (such as in elderly individuals with osteoporosis, severe deformity with significant involvement of the lesser metatarsophalangeal joint, and when spasticity is present). A review of biomechanical data, clinical studies, and surgical techniques is important for successful treatment of moderate and severe hallux valgus deformity. PMID- 18399601 TI - Spondylolisthesis and spondylolysis. AB - Spondylolisthesis is a common condition that can be managed both nonsurgically and surgically. More than 80% of children treated nonsurgically have resolution of symptoms. For those patients requiring surgical treatment, fusion in situ may provide adequate treatment for young patients. Patients with neural compression may require decompression to relieve symptoms, and fusion is also usually indicated. High-grade and degenerative spondylolisthesis require care that is unique to those conditions. Spondylolysis is a defect in the pars interarticularis that occurs in approximately 5% of the general population. Approximately 15% of individuals with a pars interarticularis lesion have progression to spondylolisthesis. PMID- 18399602 TI - Degenerative cervical spondylosis: clinical syndromes, pathogenesis, and management. AB - Degenerative changes in the cervical spinal column are ubiquitous in the adult population, but infrequently symptomatic. The evaluation of patients with symptoms is facilitated by classifying the resulting clinical syndromes into axial neck pain, cervical radiculopathy, cervical myelopathy, or a combination of these conditions. Although most patients with axial neck pain, cervical radiculopathy, or mild cervical myelopathy respond well to initial nonsurgical treatment, those who continue to have symptoms or patients with clinically evident myelopathy are candidates for surgical intervention. PMID- 18399603 TI - Slipped capital femoral epiphysis. AB - Slipped capital femoral epiphysis (SCFE) is a common adolescent hip disorder. The etiology of SCFE includes biomechanical and biochemical factors. SCFEs are classified as stable and unstable and are more common in boys than girls and in certain racial groups; most children with SCFEs are obese. Bilateral SCFEs may have a simultaneous or sequential presentation. Imaging studies show a posterior slip of the epiphysis relative to the metaphysis, seen early on lateral radiographs. The most common and effective initial treatment for stable SCFEs is in situ central single-screw fixation; other options include epiphysiodesis, and osteotomy with or without surgical dislocation of the hip. Later reconstruction options, typically reserved for the child with functional abnormalities, include proximal femoral osteotomy, or surgical dislocation of the hip with removal of metaphyseal prominent bone to remove the source of femoroacetabular impingement. Unstable SCFEs have an increased risk of osteonecrosis; the role of reduction, methods of fixation, and decompression are controversial. The natural history of untreated SCFEs is associated with the risk of progression and later degenerative joint disease. Based on treatment methods of 30 to 40 years ago, in situ fixation provided the best long-term function with the lowest risk of complications and the most effective delay of degenerative arthritis regardless of the severity of the SCFE. Newer technologies and techniques are allowing the reevaluation of the role of either acute or later reconstructive osteotomy. It has not yet been determined if these improved techniques will result in better outcomes than in the past. Surgical dislocation of the hip with epiphyseal orientation is a considered treatment option for those technically adept at the procedure; however, the long-term outcome compared with in situ fixation is still unknown. PMID- 18399604 TI - Subcapital realignment in slipped capital femoral epiphysis: surgical hip dislocation and trimming of the stable trochanter to protect the perfusion of the epiphysis. AB - Based on the recognition that even minor slip displacement in patients with slipped capital femoral epiphysis can regularly produce acetabular cartilage damage and early clinical symptoms, subcapital realignment of the epiphysis should be considered, although a substantial risk of osteonecrosis has been reported. A modified surgical technique can be used in which the perfusion of the epiphysis via the medial femoral circumflex artery is actively protected during surgery by executing surgical dislocation of the joint and by developing a soft tissue flap consisting of the retinaculum and the external rotator muscles. This flap allows mobilization of the epiphysis within the growth plate as well as complete callus resection of the neck without stretching the retinaculum. The dislocation of the head allows manual fixation of the epiphysis while curettage of the residual growth plate is performed, as well as manual reduction of the epiphysis onto the metaphysis under visual control of the retinaculum. With the head dislocated, any uncontrolled manipulation of the leg will result in less risk to the integrity of the retinaculum than would be the case if the head was reduced in the socket. PMID- 18399605 TI - Surgical management of forearm and distal radius fractures in children and adolescents. AB - Closed reduction with cast immobilization is the preferred method of treatment for most fractures of the forearm and distal radius in children and adolescents. Nonunion of these fractures is exceptionally rare, and remodeling restores alignment for minor incomplete reductions. Most closed reductions can be performed in the emergency department with the patient sedated or with regional anesthesia. However, reduction in an operating room with the patient under general anesthesia lowers the threshold for surgical stabilization to avoid the need for repeat reductions under general anesthesia. Surgical management with fixation is often indicated for unstable fractures, open fractures, refractures, and in circumstances involving multiple trauma and other complex injuries. PMID- 18399606 TI - Surgical treatment of carpal and hand injuries in children. AB - Most carpal and hand injuries in children are treated nonsurgically. However, surgical treatment is often required for certain clinical situations. Complications resulting from pediatric hand fractures, dislocations, and soft tissue injuries are most commonly caused by a failure to identify and treat an injury requiring surgery. PMID- 18399607 TI - Biceps tendon and superior labrum injuries: decision making. AB - Although the anatomy of the biceps tendon and the restraining structures within the rotator interval are well defined, biceps function is not clearly understood. Biceps pathology is often associated with rotator cuff disease. Although careful clinical examinations along with diagnostic testing can accurately identify patients with biceps pathology, arthroscopy is extremely valuable in the diagnosis and treatment of biceps pathology. Surgical treatment options for biceps pathology include decompression, debridement, tenotomy, and tenodesis. Several factors must be considered in this decision. The most important factors when deciding between tenodesis or tenotomy are the activity expectations of the patient, cosmesis, patient compliance, associated pathology, and patient age. Those older than 60 years tolerate a tenotomy with the fewest adverse effects. Various arthroscopic tenodesis techniques exist, including an interference screw in bone, suture anchor fixation, and suture to adjacent tissue fixation. An open subpectoral tenodesis is another option and appropriate for a retracted biceps rupture or when the biceps disease extends distal to the bicipital groove. A superior labrum anterior and posterior (SLAP) lesion at the attachment site of the biceps tendon to the superior glenoid labrum is uncommon. Clinically significant SLAP lesions are found in about 5% of all shoulder arthroscopies and may be mistaken for normal superior labral variations. Clinical examinations and diagnostic imaging tests for SLAP lesions are often unreliable, and the ultimate diagnostic confirmation is made by arthroscopy. Surgical treatment is focused on the reattachment of the unstable biceps-labral complex. PMID- 18399608 TI - Sports injuries in women: sex- and gender-based differences in etiology and prevention. AB - There has been a significant increase in the participation of women in sports at all levels, especially after the enactment of the Title IX Education Amendment in 1972. This increased participation at all levels has resulted in more women sustaining sports injuries. Data on sex- and gender-based differences in all organ systems, including the musculoskeletal system, are beign gathered. It is important to review some of the areas of sex- and gender-based differences in sports injuries for which there is significant research, such as osteoporosis, the female athlete triad, and anterior cruciate ligament injuries. It is also necessary to examine those areas in which more information is needed, such as injuries to the shoulder, foot, and ankle. PMID- 18399609 TI - Cartilage repair procedures: clinical approach and decision making. AB - Articular cartilage lesions present a clinical challenge, but treatment options for these lesions are changing and expanding. The size of the lesion, the physical demands of the patient, and treatment history all are important factors to consider when selecting a surgical approach. It is important for the surgeon to understand the physiology of the selected cartilage repair method and to be aware of its effects on the postoperative rehabilitation program. Attention to surgical technique and selection of the most appropriate rehabilitation protocol will provide patients with the best chance of optimal results. PMID- 18399610 TI - Articular cartilage repair using a resorbable matrix scaffold. AB - The creation of cartilage repair tissue relies on the implantation or neosynthesis of cartilage matrix elements. One cartilage repair strategy involves the implantation of bioabsorbable matrices that immediately fill a chondral or osteochondral defect. Such matrices support the local migration of chondrogenic or osteogenic cells that ultimately synthesize new ground substance. One such matrix scaffold, a synthetic resorbable biphasic implant (TruFit Plug; Smith & Nephew, San Antonio, TX), is a promising device for the treatment of osteochondral voids. The implant is intended to serve as a scaffold for native marrow elements and matrix ingrowth in chondral defect repair. The device is a resorbable tissue regeneration scaffold made predominantly from polylactide coglycolide copolymer, calcium sulfate, and polyglycolide. It is approved in Europe for the treatment of acute focal articular cartilage or osteochondral defects but is approved by the US Food and Drug Administration only for backfill of osteochondral autograft sites. Preclinical studies demonstrated restoration of hyaline-like cartilage in a goat model with subchondral bony incorporation at 12 months. Early clinical results of patients enrolled in the Hospital for Special Surgery Cartilage Registry have been favorable, with a good safety profile. PMID- 18399611 TI - Metabolic bone disease: a review and update. AB - Understanding the structure and formation of bone and the metabolic diseases that cause intrinsic biochemical alterations and ultimate damage to the skeletal system is an essential part of orthopaedic education and knowledge. Metabolic bone diseases such as rickets, osteomalacia, renal osteodystrophy, hyperparathyroidism, and osteoporosis often lead to subtle alterations in the patient's clinical status and to severe and disabling changes in the patient's bone structure. It is essential that orthopaedists recognize these conditions, provide a correct diagnosis, and use appropriate preventive and therapeutic treatments. PMID- 18399612 TI - Osteoporosis: the basics and case-based advanced treatment update for the orthopaedic surgeon. AB - When fragility fractures occur, urgent treatment is needed to reduce the risk of refracture. An extensive case-based review is used to provide an update on the basics of osteoporosis and the evaluation and treatment of patients with fragility fractures, with the goal of decreasing the risk of subsequent fracture. The pathophysiology of osteoporosis, mechanisms of action of optimal antifracture medications that will become a part of the pharmacologic armamentarium of every orthopaedic surgeon, and detailed evaluation and treatment information are presented to provide orthopaedic surgeons with the information needed to optimize the bone health of their patients. PMID- 18399613 TI - Venous thromboembolic disease after total hip and knee arthroplasty: current perspectives in a regulated environment. AB - Venous thromboembolic disease is the single most common reason for readmission to the hospital following total hip and total knee arthroplasty and remains a genuine threat to the life of the patient. Nevertheless, advances in surgical procedure, anesthetic management, and postoperative convalescence have altered the risks of venous thromboembolism after total joint arthroplasty in the lower extremity. Regional anesthetic techniques reduce the prevalence of venographic thrombosis by approximately 50%, and intraoperative monitoring has identified preparation of the femoral canal as the sentinel event that activates the coagulation cascade by the intravasation of marrow fat into the systemic circulation. Prevention of venographic thrombosis is most efficacious by administering fractionated heparin followed by warfarin; warfarin (international normalized ratio 2.0) appears to have a greater safety margin than fractionated heparin based on clinically meaningful bleeding events. Prevention of readmission events, proximal thrombosis, or pulmonary embolism has been demonstrated by using low-intensity warfarin. Aspirin, when used in conjunction with hypotensive epidural anesthesia after hip arthroplasty and regional anesthesia after knee arthroplasty, combined with pneumatic compression devices, also has been suggested to prevent clinical venous thromboembolism, as measured by readmission events. Oral thrombin inhibitors hold promise, but instances of liver toxicity have precluded approval in North America to date. Mechanical compression devices enhance venous flow and increase fibrinolytic activity in the lower extremity; clinical trials demonstrate efficacy in reducing venographic thrombosis alone after total knee arthroplasty and in combination with other chemoprophylactic agents after total hip arthroplasty. Extended chemoprophylaxis for 3 to 6 weeks after surgery is prudent in view of the protracted risk of thrombogenesis and the late occurrence of readmission for venous thrombosis and pulmonary embolism. PMID- 18399614 TI - Controversies in lower extremity amputation. AB - Using the experience gained from taking care of World War II veterans with amputations, Ernest Burgess taught that amputation surgery is reconstructive surgery. It is the first step in the rehabilitation process for patients with an amputation and should be thought of in this way. An amputation is often a more appropriate option than limb salvage, irrespective of the underlying cause. The decision making and selection of the amputation level must be based on realistic expectations with regard to functional outcome and must be adapted to both the disease process being treated and the unique needs of the patient. Sometimes the amputation is done as a life-saving procedure in a patient who is not expected to walk, but more often it is done for a patient who should be able to return to a full, active life. When considering amputation, the physician should establish reasonable goals when confronted with the question of limb salvage versus amputation, understand the roles of the soft-tissue envelope and osseous platform in the creation of a residual limb, understand the method of weight bearing within a prosthetic socket, and determine whether a bone bridge is a positive addition to a transtibial amputation. PMID- 18399615 TI - Malignant bone tumors. AB - Malignant bone tumors represent a small percentage of cancers nationwide and also are much less common than malignant soft-tissue tumors. The rarity of the condition makes it imperative that orthopaedic surgeons in nononcologic practices are able to recognize the symptoms that suggest a possible bony malignancy to avoid inappropriate or delayed treatment. The most common primary malignant bone tumors, osteosarcoma and Ewing's sarcoma, occur in childhood. Chondrosarcoma occurs more frequently in older adults. Rare tumors such as chordoma and adamantinoma have anatomic predilections for the sacrum and tibia, respectively. The primary symptom of a patient with a malignant bone tumor is pain, which often occurs at rest or at night. There are also characteristic findings on physical examination such as swelling or decreased joint range of motion. Patients with a likely malignancy require thorough staging to determine the extent of disease and a well-planned biopsy for accurate diagnosis. The biopsy can be an image-guided needle biopsy or an open incisional biopsy. Knowledge of specific tumor characteristics and treatment options for osteosarcoma, Ewing's sarcoma, chondrosarcoma, malignant fibrous histiocytoma, chordoma, and adamantinoma is important. Patients with osteosarcoma and resectable Ewing's sarcoma are treated with chemotherapy followed by surgical resection. Secondary sarcomas can occur in previously benign bone lesions and require aggressive treatment. Specific techniques are available for the resection of malignant bone tumors from the upper extremities, lower extremities, pelvis, and spine. Reconstruction options include the use of allografts, megaprostheses, and vascularized autografts. There has been a trend toward more prosthetic reconstructions because of early complications with allografts. The care of patients with primary malignant bone tumors requires a multidisciplinary approach to treatment. The orthopaedic oncologist is a vital member of a team composed of musculoskeletal radiologists and pathologists, radiation oncologists, medical and pediatric oncologists, and microvascular surgeons. PMID- 18399616 TI - Computer-assisted surgery: basic concepts. AB - Computer-assisted surgery has been advocated as a significant enabling technology that will enhance the surgical technique of various orthopaedic procedures. The computer becomes a sophisticated measuring tool, determining the three dimensional spatial orientation of fiducial points, which may be established by a variety of referencing methods. These fiducial points or arrays may define a bone, an instrument, or a prosthesis. Current referencing methods include using segmented computer tomograms; fluoroscopic images; ultrasound images; and imageless, direct anatomic point-picking methods. Tracking technologies use optical cameras and electromagnetic coils. Optical systems have high reliability with errors of less than 0.5 mm. Electromagnetic trackers have a similar capability, but are less reliable because of the distortion of the electromagnetic signal that may result from the complex operating room environment. Accuracy with current CT-referenced systems approximates 1 degrees or 1 mm. Other methods such as fluoroscopy or ultrasound are less precise because of difficulty related to the ability to consistently define a specific anatomic structure. Descriptive measures of outcome include standard deviation and quantification of error. Process capability indices or Six Sigma are suitable methods for comparing outcomes with computer-assisted surgery and can be generalized from various approaches. PMID- 18399617 TI - Surgical navigation in adult reconstruction surgery: techniques and clinical experience. AB - A surgeon's first response to the concept of computer-assisted orthopaedic surgery may be a sense of lost autonomy. However, a system need not and should not be designed to this end. Using the computational ability of the computer system to see beyond the human eye to view the knee with full kinematic dimensions, surgeons have recently made progress in the areas of computer assisted ligament balancing and in using smart tools for minimally invasive surgery. Full comprehension of the use of any navigational system must begin with the understanding that the system can provide feedback that is based on only specifically programmed computer code. In referring to or locating a point or axis, the computer programmer must create an absolutely reliable methodology for determining that point or axis. Expecting the computer to achieve certain functionalities when physicians have no ironclad method to achieve such functionalities exposes the true limitations of any computer-assisted process. Although the computer generates a methodology, the process of computer-assisted surgery requires that the surgeon be constantly vigilant in analyzing the feasibility of these responses. Those instrumental in the development and implementation of computer-assisted surgical techniques must ensure that measurements are valid, precise, and reproducible across subjects and users. Prospective users of computer-assisted techniques must ensure that each of these issues has been addressed before agreeing to use the system in standard practice. Once due consideration has been given to all aspects of use, and the limitations of the system are known, the benefits of computer assistance are easily understood. PMID- 18399618 TI - Computer-assisted total hip navigation. AB - Accurate implant placement in total hip replacement is important in avoiding dislocation, impingement, and edge-loading throughout the patient's postoperative functional range of motion. Current implants and bearing surfaces now provide the potential for prolonged longevity of the reconstruction, which can be compromised by malposition of the components outside of designated "safe zones." Computer assisted hip navigation offers the potential for more accurate placement of hip components and control of leg length and offset. Systems are now available that allow registration of the bony anatomy based on preoperative CT images, intraoperative fluoroscopic images, or imageless techniques based on palpation of the landmarks. In each of these approaches, cup position has been based on coordinate systems formed by identification of the anterior pelvic frontal plane. All systems have shown improved accuracy of acetabular cup placement compared with conventional manual techniques. Cup anteversion is less accurate than cup abduction with the imageless approach. Measurements made with the use of navigation systems also have shown a large variation in pelvic tilt or pelvic flexion-extension in series of cases, which can affect the appropriate cup position for each patient. The results of computer-assisted navigation in the future may be improved by incorporation of measurements of each patient's pelvic tilt, femoral stem position, and hip kinematics. PMID- 18399619 TI - The current status of computer-assisted high tibial osteotomy, unicompartmental knee replacement, and revision total knee replacement. AB - Navigation systems aid in achieving precision and accuracy of surgical steps. Conventional techniques fail to achieve optimal correction in more than 20% of patients undergoing high tibial osteotomy, which is sensitive to the accuracy of limb axis correction. A computer navigation system with dedicated software has been developed to allow the leg axis to be navigated intraoperatively, and the desired correction can be obtained with a corrective device of the appropriate height. Unicompartmental knee replacement also is sensitive to the accuracy of component placement. Standard navigation software has been modified to allow better accuracy when using a minimally invasive surgical approach. Revision total knee replacement is a challenging procedure because most of the standard bony and ligamentous landmarks are lost during the initial implantation. Using the standard navigation technique allows easier restoration of the joint line, adequate limb axis correction, and improved ligamentous stability than can be achieved using a conventional technique. However, standard software does not allow navigation of the stem extensions, which can lead to significant malpositioning of the prosthetic components. Specific software is currently being developed to address this issue. PMID- 18399620 TI - Selecting and starting an orthopaedic surgery practice. AB - Every new surgeon is faced with the same question as their residency or fellowship draws to a close: What is next? Few residents or fellows are as well prepared to answer that question as they could be. Most programs do not teach residents how to choose a practice type and location. After formal orthopaedic training, new surgeons must make decisions about their careers that can be nearly as complex and difficult as the decisions they make in the operating room. Career choices have both significant and long-term effects on the physician's financial situation, career satisfaction, and personal life. The physician should be aware of key non academic issues that arise when completing a residency program or just beginning the practice of orthopaedic surgery. PMID- 18399621 TI - The electronic medical office: optimizing solutions. AB - Optimizing the care for patients in the orthopaedic clinical setting involves a wide range of issues. Surgical techniques, preoperative and postoperative care, long-term outcomes follow-up, continuing education, and patient communication are a few of the important areas that surgeons deal with on a regular basis. Successful management of this information has an impact on clinical outcomes, direct patient care, financial decisions, and management of the surgeon's time. The development of a comprehensive electronic medical office is a powerful and probably necessary tool to successfully manage such information and achieve the goals of an effective and safe orthopaedic practice. PMID- 18399622 TI - Engineering the temporal response of photoconductive photodetectors via selective introduction of surface trap states. AB - Photoconductive photodetectors fabricated using simple solution-processing have recently been shown to exhibit high gains (>1000) and outstanding sensitivities ( D* > 10(13) Jones). One ostensible disadvantage of exploiting photoconductive gain is that the temporal response is limited by the release of carriers from trap states. Here we show that it is possible to introduce specific chemical species onto the surfaces of colloidal quantum dots to produce only a single, desired trap state having a carefully selected lifetime. In this way we demonstrate a device that exhibits an attractive photoconductive gain (>10) combined with a response time ( approximately 25 ms) useful in imaging. We achieve this by preserving a single surface species, lead sulfite, while eliminating lead sulfate and lead carboxylate. In doing so we preserve the outstanding sensitivity of these devices, achieving a specific detectivity of 10(12) Jones in the visible, while generating a temporal response suited to imaging applications. PMID- 18399623 TI - Zinc(II)-coordinated oligotyrosine: a new class of cell penetrating peptide. AB - A new series of cell penetrating peptides (CPPs) are described. The peptides are oligomers of Tyr-ZnDPA, a tyrosine derivative with an appended 2,2' dipicolylamine unit that forms a very stable coordination complex with a zinc (II) cation. This in turn allows reversible association with a chelating oxyanion such as a carboxylate or phosphate derivative. The peptide oligomers (Tyr-ZnDPA) n where n = 1, 2, 4, 8, are highly water soluble, but upon association with fatty acids or phospholipids they partition into an organic octanol phase. Furthermore, a fluorescent, fluorescein-labeled version of the octamer, (Tyr-ZnDPA) 8-Fl, can enter living mammalian cells via endocytosis and a biotin derivative can deliver fluorescein-labeled streptavidin. Fluorescence microscopy and flow cytometry experiments show that cell uptake is diminished by conditions that inhibit endocytosis. Additionally, uptake of (Tyr-ZnDPA) 8-Fl is greater than fluorescein labeled octaarginine (Arg 8-Fl) in all cell lines tested (CHO, COS-7, HeLa). Another difference with Arg 8-Fl is that cell uptake of (Tyr-ZnDPA) 8-Fl does not require the presence of heparan sulfate proteoglycans on the cell surface. This difference may eventually be of practical value because drug delivery systems that employ alternative endocytic mechanisms may be optimal for different cell lines or they may deliver selectively to different organelles within a cell. PMID- 18399624 TI - Imidazole substituent effects on oxidative reactivity of tripodal(imid)2(thioether)CuI complexes. AB - In the search for new bis(imidazole)thioether (BIT) copper complexes that accurately mimic the electronic and reactivity features of the CuM site of copper hydroxylase enzymes, a set of tripodal BIT ligands 4a, b- 6a, b has been synthesized that vary according to the imidazole C-(Ph or H) and N-(H or Me) substituents, as well as the position (2- or 4-) of the tripodal attachment. Corresponding [(BIT)Cu(L)](PF6) complexes 7a, b', 8a, b', and 9a', b' [L=CO (a), CH3CN (b)] have been prepared and characterized spectroscopically. The IR spectra of 7a- 9a (L=CO), specifically nu(CO), show little variation (2090-2100 cm(-1)), suggesting a similar electronic character of the Cu centers. In contrast, cyclic voltammetric analysis of these compounds (L=CH3CN) reveals quasi-reversible oxidation waves with significant variation of Epa in the range of + 0.45-0.57 V vs Fc/Fc(+), depending on the imidazole substituents. Each of the [(BIT)Cu(CH 3CN)]PF6 complexes reacts with dioxygen to form [(BIT)Cu(II) 2(mu-OH) 2](PF6)2 derivatives, 10- 12, but they vary considerably in their relative reactivity, following the same trend as the ease of their electrochemical oxidation, that is, [(2-BIT (NMe))Cu(CH 3CN)](+) ( 9b')>[(4-BIT (Ph,NMe))Cu(CH3CN)](+) ( 8b')>[(2-BIT (Ph2,NMe))Cu(CH3CN)](+) (1a')>[(4-BIT (Ph,NH))Cu(CH3CN)](+) (7b'). Thus, N-Me substitution and 4-tethering on the imidazole unit increase oxidation and oxygenation reactivity, while Ph-substitution and 2-tethering decrease reactivity. PM3 and DFT calculations are employed to analyze the relative stability, the electronic features, the Cu-CO vibrtional frequency, and the electrochemical and oxidative reactivity of the complexes. PMID- 18399625 TI - Isostructural potassium and thallium salts of sterically crowded triazenes: a structural and computational study. AB - Because of their similar cationic radii, potassium and thallium(I) compounds are usually regarded as closely related. Homologous molecular species containing either K(+) or Tl(+) are very rare, however. We have synthesized potassium and thallium salts MN3RR' derived from the biphenyl- or terphenyl-substituted triazenes Tph2N3H (1a), Dmp(Mph)N3H (1b), Dmp(Tph)N3H (1c), and (Me4Ter)2N3H (1d) (Dmp=2,6-Mes 2C6H3 with Mes=2,4,6-Me3C6H2; Me4Ter=2,6-(3,5-Me2C6H3)2C6H3; Mph=2 MesC6H4; Tph=2-TripC6H4 with Trip=2,4,6-(i)Pr3C6H2). The potassium complexes 2a- d were obtained in almost quantitative yield from the reaction of 1a- d with potassium metal in n-heptane. Metalation of 1a- d with TlOEt afforded the thallium triazenides 3a- d in high yields. All new compounds have been characterized by (1)H and (13)C NMR spectroscopy, elemental analysis, and X-ray crystallography and for selected species by melting point (not 3b), IR spectroscopy (2a, 2d, 3a, 3c, 3d), and mass spectrometry (2a, 3c). In the solid state structures of monomeric 2a and 3a, quasi-monomeric 2b, 3b, 2c, and 3c, and dimeric 2d and 3d additional metal-eta (n)-pi-arene-interactions to the flanking arms of the biphenyl- and terphenyl groups in the triazenide ligands of decreasing hapticity n are observed. Remarkably, all homologous potassium and thallium complexes crystallize in isomorphous cells. For 2a and 3a, the nature of the M-N and M...C(arene) bonding was studied by density functional theory calculations. PMID- 18399626 TI - Cyclam kappa4 to kappa3 ligand denticity change upon mono-n-substitution with a carboxypropyl pendant arm in a ruthenium nitrosyl complex. AB - The complex fac-[Ru(NO)Cl2(kappa(3)N(4),N(8),N(11)(1-carboxypropyl)cyclam)]Cl.H2O (1-carboxypropyl)cyclam=3-(1,4,8,11-tetraazacyclotetradecan-1-yl)propionic acid) was prepared in a one pot reaction by mixing equimolar amounts of RuNOCl 3 and (1 carboxypropyl)cyclam and was characterized by X-ray crystallography, electrospray ionization tandem mass spectrometry (ESI-MS/MS), elemental analysis, NMR, and electronic and vibrational (IR) spectroscopies. fac-[Ru(NO)Cl 2(kappa(3)N(4),N(8),N(11)(1-carboxypropyl)cyclam)]Cl.H2O crystallizes in the triclinic, space group P1, No. 2, with unit cell parameters of a=8.501(1) A, b=9.157(1) A, c=14.200(1) A, alpha=72.564(5) degrees , beta=82.512(5) degrees , gamma=80.308(5) degrees , and Z=2. The Ru-N interatomic distance and bond angle in the [Ru-NO] unit are 1.739(2) A and 167.7(2) degrees , respectively. ESI-MS/MS shows characteristic dissociation chemistry that initiates by HCl or NO loss. The IR spectrum displays a nu(NO) at 1881 cm(-1) indicating a nitrosonium character. The electronic spectrum shows absorptions bands at 264 nm (log epsilon=3.27), 404 nm (log epsilon=2.53), and 532 nm (log epsilon=1.88). (1)H and (13)C NMR are in agreement with the proposed molecular structure, which shows a very singular architecture where the cyclam ring N (with the carboxypropyl pendant arm) is not coordinated to the ruthenium resulting in a kappa(3) instead of the expected kappa(4) denticity. PMID- 18399627 TI - Modeling the resting state of oxalate oxidase and oxalate decarboxylase enzymes. AB - In view of the biological and commercial interest in models for Oxalate Decarboxylases (OxDC) and Oxalate Oxidases (OxOx), we have synthesized and characterized three new Mn (II) complexes ( 1- 3) employing N3O-donor amino carboxylate ligands (TCMA, 1,4,7-triazacyclononane- N-acetic acid; K (i) Pr 2TCMA, potassium 1,4-diisopropyl-1,4,7-triazacyclononane- N-acetate; and KBPZG, potassium N,N-bis(3,5-dimethylpyrazolyl methyl)glycinate). These complexes were characterized by several techniques including X-ray crystallographic analysis, X band electron paramagnetic resonance (EPR), electrospray ionization mass spectrometry (ESI-MS), and cyclic voltammetry. The crystal structures of 1 and 3 revealed that both form infinite polymeric chains of Mn (II) complexes linked by the pendant carboxylate arms of the TCMA (-) and the BPZG (-) ligands in a syn antipattern. Complex 2 crystallizes as a mononuclear Mn (II) cation, six coordinate in a distorted octahedral geometry. Although complexes 1 and 3 crystallize as polymeric chains, all compounds present the same N3O-donor set atoms around the metal center as observed in the crystallographically characterized OxDC and OxOx. Moreover, complex 2 also contains two water molecules coordinated to the Mn center as observed in the active site of OxDC and OxOx. ESI-MS spectrometry, combined with EPR, were useful techniques to establish that complexes 1- 3 are present as mononuclear Mn (II) species in solution. Finally, complexes 1- 3 are able to model the resting state active sites, with special attention focused on complex 2 which provides the first exact first coordination sphere ligand structural model for the resting states of both OxDC and OxOx. PMID- 18399628 TI - Highly selective synthesis of catalytically active monodisperse rhodium nanocubes. AB - Monodisperse sub-10 nm Rh nanocubes were synthesized with high selectivity (>85%) by a seedless polyol method. The {100} faces of the Rh NCs were effectively stabilized by chemically adsorbed Br- ions from trimethyl(tetradecyl)ammonium bromide (TTAB). This simple one-step polyol route can be readily applied to the preparation of Pt and Pd nanocubes. Moreover, the organic molecules of PVP and TTAB that encapsulated the Rh nanocubes did not prevent catalytic activity for pyrrole hydrogenation and CO oxidation. PMID- 18399629 TI - Size-selective Lewis acid catalysis in a microporous metal-organic framework with exposed Mn2+ coordination sites. AB - Treatment of selected aldehydes and ketones with cyanotrimethylsilane in the presence of the microporous metal-organic framework Mn3[(Mn4Cl)3BTT8(CH3OH)10]2 (1, H3BTT = 1,3,5-benzenetristetrazol-5-yl) leads to rapid conversion to the corresponding cyanosilylated products. The transformation is catalyzed by coordinatively unsaturated Mn2+ ions that serve as Lewis acids and lead to conversion yields of 98 and 90% for benzaldehyde and 1-naphthaldehyde, the highest thus far for a metal-organic framework. Larger carbonyl substrates cannot diffuse through the pores of 1, and conversion yields are much lower for these, attesting to the heterogeneity of the reaction and its dependence on guest size. The Mukaiyama-aldol reaction, known to require much more active Lewis catalysts, is also catalyzed in the presence of 1, representing the first such example for a metal-organic framework. Conversion yields obtained for the reaction of selected aldehydes with silyl enolates reach 63%, on par with those obtained with zeolites. Size selectivity is demonstrated for the first time with this reaction through the use of larger silyl enolate substrates. PMID- 18399631 TI - The Bip method, based on the induced circular dichroism of a flexible biphenyl probe in terminally protected -Bip-Xaa*- dipeptides, for assignment of the absolute configuration of beta-amino acids. AB - An induced axial chirality of the biphenyl core of the Bip (2',1':1,2;1'',2'':3,4 dibenzcyclohepta-1,3-diene-6-amino-6-carboxylic acid) residue in the terminally protected dipeptides Boc-Bip-beta-Xaa*-OMe (beta-Xaa* = L-beta(3)-HAla, L-beta(3) HVal, L-beta(3)-HLeu, L-beta(3)-HPro, trans-(1S,2S)-ACHC, trans-(1R,2R)-ACHC, trans-(1S,2S)-ACPC, trans-(1R,2R)-ACPC) resulted in an induced circular dichroism, revealing the usefulness of the Bip method for a reliable and fast assignment of the absolute configuration of chiral beta-amino acids. Remarkably, the Bip method was also applied to the unique spin-labeled, cyclic, beta-amino acids cis/trans-beta-TOAC and trans-POAC. In particular, this study allowed the assignment of the unknown absolute configurations of the enantiomers of the latter compound. PMID- 18399632 TI - Indenyl zirconium dinitrogen chemistry: N2 coordination to an isolated zirconium sandwich and synthesis of side-on, end-on dinitrogen compounds. AB - Exposure of the isolable zirconocene sandwich compounds, (eta(5)-C5Me5)(eta(5) C9H5-1-R(1)-3-R(2))Zr (R(1) = Me, (i)Pr, (t)Bu; R(2) = Me) to one atmosphere of dinitrogen resulted in N2 coordination. X-ray diffraction and NMR spectroscopy establish that the resulting dimeric dinitrogen compounds contain an unusual mu2,eta(2)-bridging indenyl ring and a weakly activated N2 ligand. N2 coordination from the isolable zirconium sandwich compounds is extremely sensitive to the number and size of the indenyl subsituents. Compounds bearing two [(i)Pr] or three methyl substituents are stable as eta(9) sandwich compounds for weeks under dinitrogen likely due to the inability to dimerize through a two atom N2 bridge. Performing the reduction of (eta(5)-C5Me5)(eta(5)-C9H5-1-R(1)-3 R(2))ZrCl2 (R(1) = (i)Pr, (t)Bu; R(2) = Me; R(1) = R(2) = SiMe3) under an N2 atmosphere produced a different outcome; rare examples of side-on, end-on zirconium dinitrogen compounds were isolated and in one case, crystallographically characterized. Protonolysis studies with weak Bronsted acids were used to evaluate the relative activation of the bridging dinitrogen ligands. PMID- 18399633 TI - Origin of the dendritic effect in multivalent enzyme-like catalysts. AB - Functionalization of multivalent structures such as dendrimers and monolayer passivated nanoparticles with catalytically active groups results in very potent catalysts, a phenomenon described as the positive dendritic effect. Here, we describe a series of peptide dendrons and dendrimers of increasing generation functionalized at the periphery with triazacyclononane, a ligand able to form a strong complex with Zn(II). Kinetic studies show that these metallodendrimers very efficiently catalyze the cleavage of the RNA model compound HPNPP, with dendrimer D32 exhibiting a rate acceleration of around 80,000 (kcat/k(uncat)) operating at a concentration of 600 nM. A theoretical model was developed to explain the positive dendritic effect displayed by multivalent catalysts in general. A detailed analysis of the saturation profile and the Michaelis-Menten parameters kcat and KM shows that it is not necessary to ascribe the positive dendritic effect to, for instance, changes in the catalytic site, increased substrate binding constant, or changes in the microenvironment. Rather it appears that the efficient catalytic behavior of multivalent catalysts is mainly determined by two factors: the number of catalytic sites occupied by substrate molecules under saturation conditions, and the efficiency of the multivalent system to generate catalytic sites in which multiple catalytic units act cooperatively on the substrate. PMID- 18399634 TI - Flexible graphene films via the filtration of water-soluble noncovalent functionalized graphene sheets. AB - Flexible graphene films were prepared by the filtration of water-soluble noncovalently functionalized graphene sheets with pyrenebutyrate. The work presented here will not only open a new way for preparing water-soluble graphene dispersions but also provide a general route for fabricating conducting films based on graphene. PMID- 18399635 TI - Synthesis of large quantities of single-walled aluminogermanate nanotube. AB - A simple aqueous synthesis yielded about 100 times more structurally well organized single-walled aluminogermanate nanotubes than previously reported "standard" procedures. The structure analyses using XRD, IRTF, TEM, and XAS were greatly facilitated by the high concentrations available, and they ascertained the imogolite-like structure of the nanotubes. Simplicity and yield of the synthesis protocol are likely to favor commercial applications of theses materials as well as simplified syntheses of other nanophases. PMID- 18399636 TI - Efficient synthesis of 2-(trifluoromethyl)nicotinic acid derivatives from simple fluorinated precursors. AB - Novel routes to 2-trifluoromethyl-nicotinic acid derivatives have been developed involving synthesis of the pyridine ring. These pyridyl compounds serve as key intermediates in the manufacture of the recently discovered COMT inhibitor, 3-(5 (3,4-dihydroxy-5-nitrophenyl)-1,2,4-oxadiazol-3-yl)-2-(trifluoromethyl)pyridine 1 oxide. PMID- 18399637 TI - Salt effect on the pH profile and kinetic parameters of microbial phytases. AB - The pH profiles of two microbial phytases were determined using four different general purpose buffers at different pH values. The roles of calcium chloride, sodium chloride, and sodium fluoride on activity were compared in these buffers. For Aspergillus niger phytase, calcium extended the pH range to 8.0. A high concentration of sodium chloride affected the activity of fungal phytase in the pH 3-4 range and shifted the pH optimum to 2.0 from 5.5 in Escherichia coli phytase. As expected, both of the microbial phytases were inhibited by sodium fluoride at acidic pH values. Because the Km for phytate increased nearly 2-fold for fungal phytase while Vmax increased about 75% in a high concentration of sodium chloride, it is possible that salt enhanced the product to dissociate from the active site due to an altered electrostatic environment. Modeling studies indicate that while the active site octapeptide's orientation is very similar, there are some differences in the arrangements of alpha-helices, beta-sheets, and coils that could account for the observed catalytic and salt effect differences. PMID- 18399638 TI - One-pot synthesis of chemically modified vegetable oils. AB - Vegetable oils are promising candidates as substitutes for petroleum base oils in lubricant applications, such as total loss lubrication, military applications, and outdoor activities. Although vegetable oils have some advantages, they also have poor oxidation and low temperature stability. One of the ways to address these issues is chemical modification of fatty acid chain of triglyceride. We report a one-pot synthesis of a novel class of chemically modified vegetable oils from epoxidized triacylglycerols and various anhydrides. In an anhydrous solvent, boron trifluoride etherate is used as catalyst to simultaneously open the oxirane ring and activate the anhydride. The reaction was monitored and products confirmed by NMR, FTIR, GPC, and TGA analysis. Experimental conditions were optimized for research quantity and laboratory scale-up (up to 4 lbs). The resultant acyl derivatives of vegetable oil, having diester substitution at the sites of unsaturation, have potential in formulation of industrial fluids such as hydraulic fluids, lubricants, and metal working fluids. PMID- 18399639 TI - Tailoring the structure of aminobisphosphonates to target plant P5C reductase. AB - Using the structure of (3,5-dichlorophenyl)aminomethylenebisphosphonic acid as a lead compound, 25 new phosphonates were synthesized and evaluated as possible inhibitors of Arabidopsis thaliana delta1-pyrroline-5-carboxylate (P5C) reductase. Derivatives substituted in the phenyl ring retained the inhibitory potential, though to a different extent. On the contrary any variation in the scaffold, i.e., the replacement of the second phosphonate moiety with a hydroxyl or an amino residue, resulted in a significant loss of biological activity. The availability of several structures capable of interfering with the catalytic mechanism in the micromolar to millimolar range allowed a proper structure activity relationship analysis, leading us to hypothesize about the steric and electronic requirements for maintenance or enhancement of the inhibitory properties. Reversal experiments with suspension cultured cells provided evidence for the occurrence of enzyme inhibition in vivo. Because in higher plants the step catalyzed by P5C reductase is shared by all pathways leading to proline synthesis, these compounds may be exploited for the design of new substances endowed with herbicidal activity. PMID- 18399640 TI - Characterization of phenolic compounds in rooibos tea. AB - Polyphenols present in rooibos, a popular herbal tea from Aspalathus linearis, were isolated in two steps. First, phenolic ingredients were separated by multilayer countercurrent chromatography (MLCCC). Preparative high-performance liquid chromatography (HPLC) was then applied to obtain pure flavonoids. The purity and identity of isolated compounds was confirmed by different NMR experiments, HPLC-diode array detector (DAD), or gas chromatography-mass spectrometry (GC-MS) analysis. This strategy proved to be valid to isolate material in up to gram quantities and to verify known and previously not published polyphenol structures. In addition the chemistry of dihydrochalcones and related intermediates was studied. The dihydrochalcone aspalathin was oxidized to the corresponding flavanone- C-glycosides (( R)/( S)-eriodictyol-6- C beta- D-glucopyranoside and ( R)/( S)-eriodictyol-8- C-beta- D-glucopyranoside). Flavanone-6- C-beta- D-glucopyranosides were further degraded to flavones isoorientin and orientin. PMID- 18399641 TI - PCB 118 and Aryl Hydrocarbon Receptor Immunoassays for Screening Dioxins in Retail Fish. AB - The efficacy of a combination of two enzyme-linked immunosorbent assay (ELISA) kits was examined for screening the toxic equivalent (TEQ) concentrations of dioxins in retail fish. The coplanar PCB-EIA system, which is a competitive immunoassay specific for polychlorinated biphenyl (PCB) 118, was tested as a screening method for mono- ortho PCBs. The Ah immunoassay (Ah-I), which is an ELISA-based aryl hydrocarbon receptor binding assay, was analyzed for its screening ability for non- ortho PCBs, polychlorinated dibenzo- p-dioxins (PCDDs), and dibenzofurans (PCDFs). Dilution and recovery tests using purified fish extracts revealed no major interference of the matrix in the PCB-EIA and suggested that the matrix effect was minimized in the Ah-I. Finally, the results for the fish samples ( n = 20) showed a strong correlation between this method and high-resolution gas chromatography coupled to high-resolution mass spectrometry for the determination of the TEQ concentrations of mono- ortho PCBs ( r = 0.99) and non- ortho PCBs and PCDD/Fs ( r = 0.97). These data indicate that our method is suitable for screening retail fish to determine the TEQ concentrations of dioxins. PMID- 18399643 TI - Microfluidic capillary system for immunoaffinity separations of C-reactive protein in human serum and cerebrospinal fluid. AB - A miniaturized system based on microfluidic capillaries is presented for point-of care testing and clinical assessment. The approach relies on microsyringe pump generated flow to deliver reagents and immunoaffinity chromatography to isolate the antigen from biological matrixes. Capillary sandwich immunoassays for C reactive protein (CRP) were demonstrated in human serum and cerebrospinal fluid (CSF), which are relevant matrixes for cardiovascular disease risk and meningitis research, respectively. Capillaries packed with antibody-coated silica beads were used to capture CRP from the matrix and a second, dye-labeled antibody was introduced to form a sandwich complex. An acidic elution buffer dissociated the antibody-antigen complexes, and the labeled antibody was detected with diode laser-induced fluorescence. Four parameter logistic functions and % relative error plots were used to model and assess the data. The calibration ranges for CRP were 0.05-3.0 microg/mL in 1:10 diluted serum and 0.01-30 microg/mL in undiluted CSF. The microfluidic apparatus employed a flow rate of 2 microL/min and a sample injection volume of 250 nL. Since it was not necessary to reach antibody-antigen reaction equilibrium and the assay platform dimensions were minimal, run times were as short as 10 min. PMID- 18399642 TI - Chitosan fibers: versatile platform for nickel-mediated protein assembly. AB - Fibers are a versatile platform because standard methods are available for the hierarchical assembly of individual fibers into controllable patterns (e.g., fabrics). Here, we report a method to biofunctionalize individual fibers by the reversible binding of proteins, and we suggest the potential of fiber assemblies by generating simple multifiber structures. Specifically, we use chitosan fibers and show that nickel can mediate assembly of histidine-tagged proteins to these fibers. Initial studies with the model His-GFP demonstrate the concept of nickel mediated protein assembly. Subsequent studies with a His-tagged streptococcal antibody-binding protein (protein G) demonstrate the assembly of antibodies to generate antibody-presenting fibers. Antibody assembly onto the fiber was shown to be controllable, and antigen-binding to these antibody-presenting fibers was measured. Importantly, antibody and antigen were observed to penetrate substantially into the individual fibers (tens of microns) to allow the assembly of pmole levels of protein per cm of fiber length. Finally, antibody-presenting fibers with different specificities were assembled into simple one- and two dimensional structures, and individual fibers in these fiber assemblies were observed to capture their respective antigens from antigen mixtures. The potential of fiber assemblies for multiplexed analysis is discussed. PMID- 18399644 TI - Performance of combinatorial peptide libraries in capturing the low-abundance proteome of red blood cells. 1. Behavior of mono- to hexapeptides. AB - For a better understanding of the behavior of solid-phase combinatorial peptide ligands for capturing the red blood cell low-abundance soluble proteome, combinatorial peptides of different lengths from a single amino acid up to a hexapeptide were evaluated. A red blood cell lysate (6 g total protein) was loaded in a cascade fashion to the six columns, which were individually eluted with 8 M urea, 2% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (v/w), and 50 mM citric acid. Each eluate was analyzed via sodium dodecyl sulfate polyacrylamide gel electrophoresis, two-dimensional maps, and nanoLC-MS/MS. THE RESULTS: mixed beads with a single amino acid attached showed the capture of a non-negligible portion of the proteome. A progressive increasing of the length of the peptide bait enlarges the pool of captured proteins. Above a length of four amino acids, a plateau is progressively reached, suggesting that not much could be gained with baits longer than six amino acids. Interestingly, whereas the beads laden with a single amino acid seem to be able to capture large-size proteins (>40 kDa), beads with progressively longer peptides capture additional proteins in the smaller size range (10-50 kDa). This suggests that interactions already begin with a single amino acid, but selectivity requires baits of proper length, at least above four amino acids. Plain beads, with a spacer arm carrying a primary amino terminal group for anchoring the baits, are essentially unable to capture proteins, suggesting that the peptide baits do not act by a mechanism of ion exchange but rather via a complex mixed mode, yielding a specific capture. PMID- 18399645 TI - Bulged adenosine influence on the RNA duplex conformation in solution. AB - The RNA single bulge motif is an unpaired residue within a strand of several complementary base pairs. To gain insight into structural changes induced by the presence of the adenosine bulge on RNA duplex, the solution structures of RNA duplex containing a single adenine bulge (5'-GCAGAAGAGCG-3'/5'-CGCUCUCUGC-3') and a reference duplex with all Watson-Crick base pairs (5'-GCAGAGAGCG-3'/5' CGCUCUCUGC-3') have been determined by NMR spectroscopy. The reference duplex structure is a regular right-handed helix with all of the attributes of an A-type helix. In the bulged duplex, single adenine bulge stacks into the helix, and the bulge region forms a well-defined structure. Both structures were analyzed by the use of calculated helical parameters. Distortions induced by the accommodation of unpaired residue into the helical structure propagate over the entire structure and are manifested as the reduced base pairs inclination and x-displacement. Intrahelical position of bulged adenine A5 is stabilized by efficient stacking with 5'-neighboring residues G4. PMID- 18399647 TI - Electrophoretic motion of a spherical particle with a symmetric nonuniform surface charge distribution in a nanotube. AB - The electrophoretic motion of a spherical nanoparticle, subject to an axial electric field in a nanotube filled with an electrolyte solution, has been investigated using a continuum theory, which consists of the Nernst-Planck equations for the ionic concentrations, the Poisson equation for the electric potential in the solution, and the Stokes equation for the hydrodynamic field. In particular, the effects of nonuniform surface charge distributions around the nanoparticle on its axial electrophoretic motion are examined with changes in the bulk electrolyte concentration and the surface charge of the tube's wall. A particle with a nonuniform charge distribution is shown to induce a corresponding complex ionic concentration field, which in turn influences the electric field and the fluid motion surrounding the particle and thus its electrophoretic velocity. As a result, contrary to the relatively simple dynamics of a particle with a uniform surface charge, dominated by the irradiating electrostatic force, that with a nonuniform surface charge distribution shows various intriguing behaviors due to the additional interplay of the nonuniform electro-osmotic effects. PMID- 18399646 TI - Pyridine nucleotide complexes with Bacillus anthracis coenzyme A-disulfide reductase: a structural analysis of dual NAD(P)H specificity. AB - We have recently reported that CoASH is the major low-molecular weight thiol in Bacillus anthracis [Nicely, N. I. , Parsonage, D., Paige, C., Newton, G. L., Fahey, R. C., Leonardi, R., Jackowski, S., Mallett, T. C., and Claiborne, A. (2007) Biochemistry 46, 3234-3245], and we have now characterized the kinetic and redox properties of the B. anthracis coenzyme A-disulfide reductase (CoADR, BACoADR) and determined the crystal structure at 2.30 A resolution. While the Staphylococcus aureus and Borrelia burgdorferi CoADRs exhibit strong preferences for NADPH and NADH, respectively, B. anthracis CoADR can use either pyridine nucleotide equally well. Sequence elements within the respective NAD(P)H-binding motifs correctly reflect the preferences for S. aureus and Bo. burgdorferi CoADRs, but leave questions as to how BACoADR can interact with both pyridine nucleotides. The structures of the NADH and NADPH complexes at ca. 2.3 A resolution reveal that a loop consisting of residues Glu180-Thr187 becomes ordered and changes conformation on NAD(P)H binding. NADH and NADPH interact with nearly identical conformations of this loop; the latter interaction, however, involves a novel binding mode in which the 2'-phosphate of NADPH points out toward solvent. In addition, the NAD(P)H-reduced BACoADR structures provide the first view of the reduced form (Cys42-SH/CoASH) of the Cys42-SSCoA redox center. The Cys42-SH side chain adopts a new conformation in which the conserved Tyr367' OH and Tyr425'-OH interact with the nascent thiol(ate) on the flavin si-face. Kinetic data with Y367F, Y425F, and Y367,425F BACoADR mutants indicate that Tyr425' is the primary proton donor in catalysis, with Tyr367' functioning as a cryptic alternate donor in the absence of Tyr425'. PMID- 18399648 TI - Comparing in vitro and in vivo activity of 2'-O-[2-(methylamino)-2-oxoethyl]- and 2'-O-methoxyethyl-modified antisense oligonucleotides. AB - A number of 2'- O-modified antisense oligonucleotides have been reported for their potential use in oligonucleotide-based therapeutics. To date, most of the in vivo data has been generated for 2'-O-MOE (2'-O-methoxyethyl)- and 2'-O-Me (2' O-methyl)-modified ASOs (antisense oligonucleotides). We now report the synthesis and biological activity of another 2'-O-modification, namely 2'-O-[2 (methylamino)-2-oxoethyl] (2'-O-NMA). This modification resulted in an increase in the affinity of antisense oligonucleotides to complementary RNA similar to 2' O-MOE-modified ASOs as compared to first-generation antisense oligodeoxynucleotides. The ASO modified with 2'-O-NMA reduced expression of PTEN mRNA in vitro and in vivo in a dose-dependent manner similar to 2'-O-MOE modified ASO. Importantly, toxicity parameters such as AST, ALT, organ weights, and body weights were found to be normal similar to 2'-O-MOE ASO-treated animal models. The data generated in these experiments suggest that 2'-O-NMA is a useful modification for potential application in both antisense and other oligonucleotide-based drug discovery efforts. PMID- 18399649 TI - Exploiting structure-activity relationships in docking. AB - From the perspective of 2D chemical descriptors, error in docking activity predictions is separated into noise and systematic components. This error framework explains how fitting docking scores to a 2D-QSAR equation often improves accuracy as well as its logical limits. Intriguingly, in examined cases where multiple docking models (e.g., multiple crystal structures or multiple scoring functions) are available for an enzyme, the noise component of error dominates the difference between the more accurate and less accurate docking models. When this is true, the QSAR equation fit statistics can rank each docking score set's accuracy in the absence of experimental activity data. PMID- 18399650 TI - The general synthesis and trapping of 3-substituted 1-chlorocyclopropenes. AB - Treatment of 1-bromo-2,2-dichloro-3-trimethylsilylcyclopropane with CsF in THF refluxing temperature gave chlorocyclopropenyl cation (1), which reacted with various nucleophiles to generate 3-monosubstituted 1-chlorocyclopropenes. The Diels-Alder reactions of 3-monosubstituted 1-chlorocyclopropenes with 1,3 diphenylisobenzofuran (DPIBF) yielded only the exo-anti adducts in excellent yield. When cation 1 was treated with water and trapped with DPIBF, an aldehyde, 12, was isolated. PMID- 18399651 TI - Dimethyl sulfide induced [3+2] annulation strategy: an efficient synthesis of functionalized dihydropyrazole derivatives using the Baylis-Hillman bromides. AB - Baylis-Hillman bromides have been successfully employed as a valuable source of 1,3-dipoles for cycloaddition onto dialkyl azodicarboxylates (dipolarophiles) under the influence of dimethyl sulfide and potassium carbonate to provide functionalized dihydropyrazole derivatives in a simple one-pot [3+2] annulation strategy. PMID- 18399652 TI - Diastereoselective synthesis of delta-aminoacids through domino Ireland-Claisen rearrangement and Michael addition. AB - A novel domino reaction--stereoselective Ireland-Claisen rearrangement and asymmetric Michael addition--is described. A protocol starting from Baylis Hillman adducts 3a-f using chiral lithium amide affords optically active gamma substituted delta-amino acids 4a-f with high diastereoselectivities and enantioselectivities. The acid can be isolated easily from large-scale reactions and transformed to 2,3-disubstituted piperidines 11 or 2-substituted nipecotic acid derivates 12. PMID- 18399653 TI - Utilization of a Michael addition: dipolar cycloaddition cascade for the synthesis of (+/-)-cylindricine C. AB - A new approach to the marine alkaloid (+/-)-cylindricine C has been devised. The key element of the synthesis consists of a Michael addition/dipolar cycloaddition cascade between 2,3-bis(phenylsulfonyl)-1,3-butadiene and 9 triisopropylsilanyloxy-non-1-en-5-one oxime. The resulting cycloadduct was converted into (+/-)-cylindricine C by a sequence of reactions including a reductive cyclization, intramolecular enolate alkylation, and conjugate addition to introduce the n-hexyl side chain. PMID- 18399654 TI - Modular synthesis of the C9-C27 degradation product of aflastatin A via alkyne epoxide cross-couplings. AB - A modular approach to the synthesis of complex polyketide natural products is demonstrated for the synthesis of the C9-C27 degradation product from aflastatin A. The product of the cross-coupling of C23-C27 terminal alkyne with C17-C22 epoxide underwent functionalization of the resulting internal alkyne, which was then coupled similarly with C9-C16 epoxide. This synthesis concluded with regio- and stereoselective addition of methyl onto the internal alkyne followed by stereoselective hydroboration-oxidation. PMID- 18399655 TI - A tandem approach to isoquinolines from 2-azido-3-arylacrylates and alpha diazocarbonyl compounds. AB - 2-Azido-3-arylacrylates react with alpha-diazocarbonyl compounds and triphenylphosphine to furnish isoquinolines in 60-92% yields. The tandem process involves a Wolff rearrangement, an aza-Wittig reaction, and an electrocyclic ring closure. The procedure is efficient, rapid, and general, and the substrates are readily available. PMID- 18399656 TI - Synthesis, structure, and metal complexation behavior of a new type of functionalized chiral phenanthroline derivative. AB - SmI(2) serves as an effective promoter for the coupling of 1,10-phenanthroline with an epoxide to generate a new class of chiral, functionalized ligands that readily form complexes with metals. Structural studies of the resulting phenanthroline derivative and two of its metal complexes are reported. PMID- 18399657 TI - Comparative study of the photochemistry of the azidopyridine 1-oxides. AB - The photochemistry of azidopyridine 1-oxides was studied using an array of glass and matrix isolation techniques. As with room temperature, the photochemistry of 4-azidopyridine 1-oxide is dominated by triplet nitrene chemistry. However, in the case of the 3-azide, matrix photolysis indicates the formation of diazabicyclo[4.1.0]hepta-2,4,6-triene N-oxide and diazacycloheptatetraene N-oxide intermediates as well as triplet nitrene. PMID- 18399658 TI - Improved regioselectivity in pyrazole formation through the use of fluorinated alcohols as solvents: synthesis and biological activity of fluorinated tebufenpyrad analogs. AB - The preparation of N-methylpyrazoles is usually accomplished through reaction of a suitable 1,3-diketone with methylhydrazine in ethanol as the solvent. This strategy, however, leads to the formation of regioisomeric mixtures of N methylpyrazoles, which sometimes are difficult to separate. We have determined that the use of fluorinated alcohols such as 2,2,2-trifluoroethanol (TFE) and 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) as solvents dramatically increases the regioselectivity in the pyrazole formation, and we have used this modification in a straightforward synthesis of fluorinated analogs of Tebufenpyrad with acaricide activity. PMID- 18399660 TI - Surface plasmon resonance enhanced transmission of light through gold-coated diffraction gratings. AB - Narrow peaks are observed in the transmission spectra of p-polarized light passing through a thin gold film that is coated on the surface of a transparent diffraction grating. The spectral position and intensity of these peaks can be tuned over a wide range of wavelengths by simple rotation of the grating. The wavelengths where these transmission peaks are observed correspond to conditions where surface plasmon resonance occurs at the gold-air interface. Light diffracted by the grating couples with surface plasmons in the metal film to satisfy the resonant condition, resulting in enhanced light transmission through the film. Notably, this phenomenon is not observed at flat, gold-coated surfaces or uncoated gratings, where coupling to surface plasmons does not occur. The nature of the coupling and, thus, the details of light transmission are governed by the momentum matching conditions between the diffracted light and the surface plasmons. In the presence of bound analytes or surface films, the enhanced transmission peaks are red-shifted, making a simple, yet highly responsive sensing platform. The utility of this platform is demonstrated for ex situ sensing by analyzing thin films of various thicknesses and detecting a model immunoreaction between bovine serum albumin and anti-bovine serum albumin. This grating-based transmission surface plasmonic device represents a simple and sensitive platform, which can be readily tuned to enhance performance and be used in the study of a variety of surface adsorption processes or analysis of biomolecular interactions. PMID- 18399659 TI - Capillary LC-MS for high sensitivity metabolomic analysis of single islets of Langerhans. AB - Reversed-phase, packed capillary liquid chromatography interfaced by electrospray ionization to mass spectrometry was explored as an analytical method for determination of metabolites in microscale tissue samples using single islets of Langerhans as a model system. With the use of a 75 microm inner diameter column coupled to a quadrupole ion trap mass spectrometer in full scan mode, detection limits of 0.1-33 fmol were achieved for glycoloytic and tricarboxylic acid cycle metabolites. Reproducible processing of islets for analysis with little loss of metabolites was performed by rapid freezing followed by methanol-water extraction. The method yielded 20 microL of extract of which just 15 nL was injected suggesting the potential for performing multiple assays on the same islet. Approximately 200 presumed metabolites could be detected, of which 22 were identified by matching retention times and MS/MS spectra to standards. Relative standard deviations for peak detection was from 7 to 18% and was unaffected by storage for up to 11 days. The method was used to detect changes in metabolism associated with increasing extracellular islet glucose concentration from 3 to 20 mM yielding results largely consistent with known metabolism of islets. Because most previous studies of islet metabolism have only observed a few compounds at once and require far more tissue, this measurement method represents a significant advance for studies of metabolism of islets and other microscale samples. PMID- 18399661 TI - Drug assessment based on detection of L-glutamate released from C6 glioma cells using an enzyme-luminescence method. AB - Monitoring of excitation activity of nerve cells is very useful for not only brain research but also assessment of the effects of various chemicals, including drugs and toxins. We previously reported a novel enzyme-luminescence method for real-time monitoring of l-glutamate release from C6 glioma cells with high levels of sensitivity ( approximately 10 nM) and temporal resolution (<1 s) using a luminescence plate reader. In the present study, we tested the applicability of this novel system for assessment of effects of drugs in vitro. Several drugs (e.g., veratridine and 4-aminopyridine) were administered to C6 glioma cells for inducing glutamate release. Moreover, antagonists of voltage-dependent Ca (2+) channels (e.g., nifedipine, flunarizine, and NiCl 2) and Na (+) channels (e.g., carbamazepine and lidocaine) were applied separately for evaluating the effects of these chemicals on glutamate release from the cells. The combined effect of carbamazepine and lidocaine was also investigated by using our method, and the combined effect was found to be more potent than that of single drug administration. These results indicated that the glutamate release from C6 cells was modulated by these drugs in a way similar to that found by using several conventional analytical techniques. We therefore conclude that the developed monitoring system for real-time detection of dynamic l-glutamate release from cells could be very useful for application to assessment of drugs acting on the nervous system. PMID- 18399662 TI - Development of quantitative cell-based enzyme assays in microdroplets. AB - We describe the development of an enzyme assay inside picoliter microdroplets. The enzyme alkaline phosphatase is expressed in Escherichia coli cells and presented in the periplasm. Droplets act as discrete reactors which retain and localize any reaction product. The catalytic turnover of the substrate is measured in individual droplets by monitoring the fluorescence at several time points within the device and exhibits kinetic behavior similar to that observed in bulk solution. Studies on wild type and a mutant enzyme successfully demonstrated the feasibility of using microfluidic droplets to provide time resolved kinetic measurements. PMID- 18399663 TI - Temporal evolution of the composition of mixed monolayers on TiO2 surfaces: evidence for a dimerization-induced chelate effect. AB - Mixed monolayers of octanoic acid (OA) and 16-mercaptohexadecanoic acid (MHDA) were adsorbed to nanocrystalline TiO(2) films from mixed solutions in tetrahydrofuran. For a range of solution compositions, the mole fraction of MHDA within the mixed monolayers (chi (MHDA,surf)) exceeded that of the coadsorption solution. In addition, chi (MHDA,surf) increased with time, while the sum of the surface coverages of MHDA and OA remained constant. To account for these effects, we propose a mechanism involving disulfide formation between the terminal thiol groups of surface-adsorbed MHDA molecules. Disulfide formation leads to an increase in the surface adduct formation constant ( K(ad)) of dimeric MHDA, causing the gradual displacement of OA from the surface. The mechanism is supported by spectroscopic evidence and desorption kinetics. These are the first examples of mixed monolayers that undergo time-dependent compositional changes as a result of covalent bond formation between surfactants. Our findings illustrate that dimerization and other intermolecular interactions between surfactants may dramatically influence the composition and terminal functionalization of a wide range of mixed monolayer systems. PMID- 18399664 TI - Zinc oxide growth morphology on self-assembled monolayer modified silver surfaces. AB - Using organic molecules to direct inorganic crystal growth has opened up new avenues for controlled synthesis on surfaces. Combined with soft lithography to form patterned templates, self-assembled monolayers (SAMs) have been shown to be a powerful approach for the assembly of inorganic nanostructures. In this work, we show that the surface free energy of SAM-modified silver, which depends on end groups and deposition method of SAMs, has a dramatic effect on the nucleation and growth of crystalline ZnO, a technologically important material, from supersaturated solutions. For SAMs with inert methyl end groups, ZnO nucleation is inhibited. For SAMs with chemically active (carboxylic or thiol) end groups, the ZnO morphology is found to be three-dimensional nanorods on low-surface energy surfaces and two-dimensional thin films on high-energy surfaces. PMID- 18399665 TI - Green chemistry for large-scale synthesis of semiconductor quantum dots. AB - Large-scale synthesis of semiconductor nanocrystals or quantum dots (QDs) with high concentration and high yield through simultaneously increasing the precursor concentration was introduced. This synthetic route conducted in diesel has produced gram-scale CdSe semiconductor quantum dots (In optimal scale-up synthetic condition, the one-pot yield of QDs is up to 9.6g). The reaction has been conducted in open air and at relatively low temperature at 190-230 degrees C in the absence of expensive organic phosphine ligands, aliphatic amine and octadecene, which is really green chemistry without high energy cost for high temperature reaction and unessential toxic chemicals except for Cd, which is the essential building block for QDs. PMID- 18399666 TI - Effect of metal-molecule contact roughness on electronic transport: bacteriorhodopsin-based, metal-insulator-metal planar junctions. AB - Molecular electronics is very much about contacts, and thus understanding of any generic contact effect is essential to its advance. For example, it is still not obvious in how far variations in electrode roughness of macroscopic contacts can lead to rectification. Here we report an investigation of this contact effect on electronic transport properties using metal-insulator-metal planar junctions with a 5 nm thick bacteriorhodopsin-based insulator as model system. We demonstrate that the experimentally observed rectifying behavior is not an intrinsic property of the molecules used, but rather of the local contact quality. Even a slight increase in surface roughness of the bottom electrode gives rise to distinct rectifying behavior in these and, by extrapolation, possibly other molecular junctions. PMID- 18399667 TI - Covalent and physical cross-linking of photonic crystals with 10-fold-enhanced chemomechanical stability. AB - We report opal photonic crystals that are self-assembled from functional polymer particles. We randomly copolymerized functional side-chain monomers containing motifs that form homodimers or heterobridges. These include ether or methylene bridges, hydrazone bridges, acids for anhydride formation, low- T g copolymers or physical cross-links by hydrogen bonds and/or polarity. To generate particles that are monodisperse, spherical, and functionalized, we combined emulsifier-free synthesis with swelling synthesis steps. Laser diffraction from centimeter-sized beams, white-light interferometry, and atomic force microscopy demonstrates symmetry and homogeneity across the entire crystal without the loss of interstitial volume. Compared to the stability of nonfunctional particles, the stability of the crystal against immersion in water and isopropanol was enhanced from 10 to a perfect 100%. One of the successful approaches (methylene bridges from N-methylolmethacrylamid) is triggered by thermal activation, but as shown, this is operative far from the trivial regime of sintering. We demonstrate successful infiltration with and solvation of a laser-polymerizable resin, thus enabling the processing of 3D photonic waveguide structures. PMID- 18399668 TI - Hydrothermal-induced structure transformation of polyelectrolyte multilayers: from nanotubes to capsules. AB - The assembled polyelectrolyte nanotubes composed of poly(styrenesulfonate) and poly(allylamine hydrochloride) multilayers by using the layer-by-layer assembly combined with the porous template method can be transformed into capsules by a high-temperature treatment. Scanning electron microscopy and confocal laser scanning microscopy images revealed the whole transition process. The structure transformation of polyelectrolyte multilayers after annealing can be initiated by the input of thermal energy which leads to a breakage of ion pairs between oppositely charged polyelectrolyte groups. The transition process from tubes to capsules is supposed to be driven by the Raleigh instability and leads to the generated polyelectrolyte capsules with different sizes. PMID- 18399669 TI - Controlling orientations of immobilized oligopeptides using N-terminal cysteine labels. AB - This letter reports a strategy of using N-terminal cysteine labels for controlling the immobilization of oligopeptides on aldehyde-terminated surfaces through the formation of stable thiazolidine rings. We also study the effect of cysteine position (either N-terminal or C-terminal) and lysine residue on the immobilization of oligopeptides. On the basis of our ellipsometry and quartz crystal microbalance (QCM) results, we conclude that the proposed immobilization strategy is highly site-specific. It works only when cysteine is in the N terminal position, and the formation of thiazolidine is much faster than the formation of imines between lysine residues and aldehydes, even in the presence of a reducing agent such as NaBH(3)CN. By labeling an oligopeptide CSNKTRIDEANNKATKML with an N-terminal cysteine, we immobilize this oligopeptide on an aldehyde-terminated surface and investigate the enzymatic activity of trypsin acting on the oligopeptide. It is found that trypsin is able to cleave the immobilized oligopeptide having a single anchoring point at the N-terminal cysteine. No cleavage is observed when the oligopeptide is immobilized through multiple anchoring points at lysine residues. PMID- 18399671 TI - Chemisorption of phosphoric acid and surface characterization of As passivated AlN powder against hydrolysis. AB - By simply refluxing a commercial AlN powder in a mixture solution of ethanol, H(3)PO(4), and Al(H(2)PO(4))(3) for 24 h at 80 degrees C, the powder was successfully passivated against hydrolysis. The phosphate layer formed on the surface of AlN powder was found to be quite stable toward protecting the powder from hydrolysis. The efficacy of the coating was established by suspending the treated and the untreated powders in water for 72 h and subsequently characterizing them by X-ray diffraction (XRD), Fourier transform infrared (FT IR), X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), and Raman analysis. The good dispersing behavior of the treated AlN powder in water was confirmed by the low viscosity of an AlN suspension containing 50 vol % solids demonstrating the viability of replacing organic solvents by water in colloidal processing of AlN-based ceramics. PMID- 18399670 TI - A highly stable nonbiofouling surface with well-packed grafted zwitterionic polysulfobetaine for plasma protein repulsion. AB - An ideal nonbiofouling surface for biomedical applications requires both high efficient antifouling characteristics in relation to biological components and long-term material stability from biological systems. In this study we demonstrate the performance and stability of an antifouling surface with grafted zwitterionic sulfobetaine methacrylate (SBMA). The SBMA was grafted from a bromide-covered gold surface via surface-initiated atom transfer radical polymerization to form well-packed polymer brushes. Plasma protein adsorption on poly(sulfobetaine methacrylate) (polySBMA) grafted surfaces was measured with a surface plasmon resonance sensor. It is revealed that an excellent stable nonbiofouling surface with grafted polySBMA can be performed with a cycling test of the adsorption of three model proteins in a wide range of various salt types, buffer compositions, solution pH levels, and temperatures. This work also demonstrates the adsorption of plasma proteins and the adhesion of platelets from human blood plasma on the polySBMA grafted surface. It was found that the polySBMA grafted surface effectively reduces the plasma protein adsorption from platelet-poor plasma solution to a level superior to that of adsorption on a surface terminated with tetra(ethylene glycol). The adhesion and activation of platelets from platelet-rich plasma solution were not observed on the polySBMA grafted surface. This work further concludes that a surface with good hemocompatibility can be achieved by the well-packed surface-grafted polySBMA brushes. PMID- 18399672 TI - N,N'-diarylureas: a new family of atropisomers exhibiting highly diastereoselective reactivity. AB - 2,6-disubstituted N-aryl ureas rotate slowly about their Ar-N bonds and can exist as separable atropisomers. They also react remarkably diastereoselectively, with the urea axis controlling new stereogenic centers with high fidelity in a variety of nucleophilic and electrophilic addition reactions. The sense of diastereoselectivity in lateral lithiation-electrophilic quench reactions is electrophile-dependent and appears to be the result of stereospecific reaction with one of two interconvertible diastereoisomeric organolithiums. PMID- 18399673 TI - Geometric and chelation influences on the electronic structure and optical properties of tetra(carboxylic acid)phenyleneethynylene dyes. AB - A quantum-chemical study on the consequences of geometric modification and chelation on the electronic structure and optical properties of a tetra(carboxylic acid)phenyleneethynylene dye, of interest for chemical sensing applications, is presented. Rotation within the central biphenylene and complexation with divalent metal ions--in particular Cu2+--lead to notable changes in the absorption and emission profiles. Calculations at both the density functional theory (DFT) and Hartree-Fock (HF) levels are used to evaluate geometric potential energy surfaces for rotation within the central biphenylene unit; HF coupled with configuration interaction singles (HF-CIS) is used to investigate the first excited state of the dye. Time-dependent DFT (TDDFT) calculations are employed to assess changes in optical absorption and fluorescence as a function of geometry and chelation. PMID- 18399674 TI - Low-lying electronic states and dissociation energies of the monochlorides of Cr, Mn, Fe, Co, and Ni. AB - Published equilibrium data involving the gaseous monochlorides of Cr, Mn, Fe, Co, and Ni have been re-examined by thermochemical analysis, using more recent information on the low-lying electronic states, yielding D degrees 0 values in kcal mol-1 of CrCl (90.0), MnCl (79.8), FeCl (79.3), CoCl (81.3), and NiCl (88.1). Although this revised approach is believed to yield more reliable values of the FeCl, CoCl, and NiCl dissociation energies, results show that use of M+ electronic levels in place of the adopted MCl values leads to alternate D degrees 0(MCl) values agreeing within 1.6 kcal mol-1, providing a useful check on electronic-level contributions to the thermochemical calculations. PMID- 18399675 TI - Interaction in the ternary complexes of HCl-methanol-X, X = H2O or NH3: Ab initio calculations and on-the-fly molecular dynamics. AB - Dynamics, structures, energetics, and vibrational spectra of the ternary complexes of hydrogen chloride with either methanol and water or methanol and ammonia were investigated by on-the-fly molecular dynamics and ab initio and density functional theory (DFT) with aug-cc-pvDZ basis sets. Addition of CH3OH to the HCl-NH3 system catalyzes the proton transfer from HCl to NH3. However, the dynamics of the system show that the proton is not localized on NH3; rather, it is shared between N and Cl. PMID- 18399676 TI - Coupled-cluster study of isomers of H2SO2. AB - A theoretical study has been made on six isomers of H2SO2 using coupled-cluster singles and doubles with noniterative triple excitations (CCSD(T)). The isomers studied are sulfoxylic acid (S(OH)2; C2 and Cs conformers), sulfinic acid (HS(=O)OH; 2 C1 conformers), dihydrogen sulfone (H2SO2; C2v), sulfhydryl hydroperoxide (HSOOH; C1), thiadioxirane (Cs), and dihydrogen persulfoxide (H2SOO; Cs). Molecular geometries, harmonic vibrational frequencies, and infrared intensities of all species were obtained using the CCSD(T) method and the 6 311++G(2d,2p) basis set. All aforementioned species were found to be local minima, with the exception of thiadioxirane, which has one imaginary frequency. A prior possible infrared observation of sulfinic acid was reassessed on the basis of the present data. In agreement with previous MP2 results, the present CCSD(T) data provide support for at most 4 of the 8 observed frequencies. The CCSD(T) frequencies and intensities should be of assistance in future identification of H2SO2 isomers by vibrational spectroscopy. Relative energies were calculated using the CCSD(T) method and several larger basis sets. As found previously, the lowest energy species is C2 S(OH)2, followed by Cs S(OH)2, HS(=O)OH, H2SO2, HSOOH, thiadioxirane, and H2SOO. Expanding the basis set significantly reduces the relative energies of HS(=O)OH and H2SO2. The CCSD(T) method was used with extended basis sets (up to aug-cc-pV(Q+d)Z) and basis set extrapolation in two reaction schemes to calculate the DeltaH degrees t (25 degrees C) of C2 S(OH)2. The two reaction schemes gave -285.8 and -282.7 kJ mol-1, which are quite close to a prior theoretical estimate (-290 kJ mol-1). PMID- 18399677 TI - Methyl group dynamics and the onset of anharmonicity in myoglobin. AB - The role of methyl groups in the onset of low-temperature anharmonic dynamics in a crystalline protein at low temperature is investigated using atomistic molecular dynamics (MD) simulation. Anharmonicity appears at approximately 150 K, far below the much-studied solvent-activated dynamical transition at approximately 220 K. A significant fraction of methyl groups exhibit nanosecond time scale rotational jump diffusion at 150 K. The splitting and shift in peak position of both the librational band (around 100 cm(-1)) and the torsional band (around 270-300 cm(-1)) also differ significantly among methyl groups, depending on the local environment. The simulation results provide no evidence for a correlation between methyl dynamics and solvent exposure, consistent with the hydration-independence of the low-temperature anharmonic dynamics observed in neutron scattering experiments. The calculated proton mean-square fluctuation and methyl NMR order parameters show a systematic nonlinear dependence on the rotational barrier which can be described using model functions. The methyl groups that exhibit many rotational excitations are located near xenon cavities, suggesting that cavities in proteins act as activation centers of anharmonic dynamics. The dynamic heterogeneity and the environmental sensitivity of motional parameters and low-frequency spectral bands of CH(3) groups found here suggest that methyl dynamics may be used as a probe to investigate the relation between low-energy structural fluctuations and packing defects in proteins. PMID- 18399678 TI - Scaling aspects of block co-polymer adsorption on curved surfaces from nonselective solvents. AB - In this paper, we have developed a geometric-based scaling model that describes the adsorption of diblock copolymer chains from good solvents and theta-solvents onto reactive surfaces of varying curvatures. To evaluate the impact of particle size on the adsorption process, we probed the adsorption of poly(styrene-b methymethacrylate) (PS-PMMA) diblock copolymers from solvents with different degrees of selectivity on aluminum oxide (Al(2)O(3)) surfaces belonging to particles of different sizes. When the adsorbed PMMA layer is dense enough (in the case of a theta-solvent for the PMMA block), our results show good correlation between the theory and experimental results, pointing to the formation of a PMMA adsorption layer and a brushlike PS layer. Conversely, when adsorption occurs from a nonpreferential solvent, particularly on particles with high curvature, the PMMA adsorption layer at the surface becomes less dense and the grafted PS moiety exhibits a transitional morphology consisting of several layers of increasingly sparsely spaced blobs. PMID- 18399679 TI - Method for macromolecular colocalization using atomic recombination in dynamic SIMS. AB - Localizing two or more components of assemblies in biological systems requires both continued development of fluorescence techniques and invention of entirely new techniques. Candidates for the latter include dynamic secondary ion mass spectrometry (D-SIMS). The latest generation of D-SIMS, the Cameca NanoSIMS 50, permits the localization of specific, isotopically labeled molecules and macromolecules in sections of biological material with a resolution in the tens of nanometers and with a sensitivity approaching in principle that of a single protein. Here we use two different systems, crystals of glycine and mixtures of proteins, to show that the formation of recombinant CN secondary ions under Cs bombardment can be exploited to create a new colocalization technique. We show experimentally that the formation of the recombinant (13)C(15)N secondary ion between (13)C- and (15)N-labeled macromolecules is indeed an indicator of the distance between the interacting macromolecules and on their shape. We build up a convolution model of the mixing-recombination process in D-SIMS that allows quantitative interpretations of the distance-dependent formation of the recombinant CN. Our results show that macromolecules can be colocalized if they are within 2 nm of one another. We discuss the potential advantages of this new technique for biological applications. PMID- 18399680 TI - Two-dimensionally well-ordered multilayer structures in thin films of a brush polypeptide. AB - In this study, we report the first production of two-dimensionally well-ordered molecular multilayers (i.e., with a well-defined molecular lamellar structure) based on the antiparallel beta-sheet chain conformation in thin films of a brush polypeptide, poly(S-n-hexadecyl-dl-homocysteine) (PHHC), through the use of a simple spin-coating process and the quantitative structural and property analysis of the thin films using a grazing incidence X-ray scattering technique combined with Fourier transform infrared spectroscopy and differential scanning calorimetry. These analyses provide detailed information about the structure and molecular conformation of the self-assembled lamellae in the PHHC thin film, which is not easily obtained using conventional techniques. Moreover, we used the in situ measurements carried out at various temperatures and the data analyses to establish mechanisms for the evolution of the self-assembled lamellar structures in the film and for their melting. In addition, we propose molecular structure models of the PHHC polymer molecules in the thin film at various temperatures. PMID- 18399681 TI - Quantized hydration energies of ions and structure of hydration shell from the experimental gas-phase data. AB - With previous data on alkali metal and halide ions included [Rais, J.; Okada, T. Anal. Sci. 2006, 22, 533], we analyzed rather broad data on ionic hydration from the point of view of gaseous cluster energetics. We have now added alkaline earth cations, Zn(2+), H(+), OH(-), Cu(+), Ag(+), Bi(+), Pb(+), and alkylammonium cations. The present analysis revealed the octa-coordinated nature of alkaline earth cations, which is not fully pronounced for Be(2+) and Zn(2+), existence of Eigen protonium complex, which is trigonally hydrated, and particular property of the first OH-, H(2)O cluster. Whereas these findings are generally in accordance with theoretical model calculation studies, we have foreseen in addition tetrahedral hydration for halide anions and Rb(+) and Cs(+), as well as for alkylammonium ions. The obtained picture of the quantized solvation of ions is mirrored in the ionization potentials of outer electrons of pertinent atoms. This is a second independent phenomenon, and together, they invoked a common pattern formation ("Aufbau") obeying tetra- and octa-coordinated principles. PMID- 18399682 TI - An efficient method for calculating atomic charges of peptides and proteins from electronic populations. AB - An efficient method is presented to calculate atomic charges of peptides and proteins derived from Mulliken electronic populations for terminally blocked amino acids (Ac-X-NHMe, X = any neutral or charged residue) calculated at the B3LYP/6-31G(d,p)//HF/6-31G(d,p) level of theory. This electronic population derived atomic charges (EPAC) method is based on the geometry-dependent atomic charge (GDAC) method proposed by Cho et al. (J. Phys. Chem. B 2001, 105, 3624), in which atomic charges are calculated by using the partial equalization of atomic electronegativities with electronegativity parameters and damping factors given by interatomic distances between covalently bonded atoms in a molecule. The overall mean absolute difference (mad) and root-mean-square deviation (rmsd) between dipole moments micro(EPAC) and micro(B3LYP), obtained from EPAC charges and from B3LYP/6-31G(d,p) level calculations, respectively, for Ac-X-NHMe are estimated to be 0.38 and 0.59 D, respectively, for 26 representative conformations in the training set, and 0.54 and 0.79 D, respectively, for 172 representative conformations not used for parametrization. For Ac-(Ala)(n)-NHMe (n = 2-6), the EPAC method reasonably predicts the increase of the dipole moment with increase of the chain length, although the deviations from the micro(B3LYP) values are somewhat larger. For Ac-Ala-NHMe and Ac-(Ala)(3)-NHMe, the EPAC charge for a specific type of atom does not depend on its position in the sequence or on the length of the sequence. In addition, charge neutrality holds for any Ala residue of these two peptides. Thus, these results suggest that the EPAC charges derived from B3LYP/6-31G(d,p) Mulliken populations can be used reliably for conformational analysis of peptides and proteins. PMID- 18399683 TI - Microemulsion polymerizations via high-frequency ultrasound irradiation. AB - The synthesis of nanosized polymer latex particles using high-frequency ultrasound (213 kHz) has been successfully performed. The effects of surfactant type and concentration of surfactants on the rates of polymerization, latex size, and molecular weights of the polymers produced are presented. PMID- 18399684 TI - Metal cation induced cubic phase in poly(ethylene glycol)-functionalized dioleoylphosphatidylethanolamine aqueous dispersions. AB - Metal cations (Mn(2+) or Ca(2+)) in aqueous dispersions of mixtures of dioleoylphosphatidylethanolamine (DOPE) and poly(ethylene glycol)-functionalized DOPE (DOPE-PEG(350)) induce, above a certain amount of the PEG lipid component, a phase transition from the inverted hexagonal phase H(II) to the bicontinuous inverted cubic phase Q(224) with space group Pn3m. The process is driven by the decrease of free elastic energy due to the Gaussian curvature of the cubic phase. The structural characterization of the phase behavior over the whole explored range of DOPE-PEG/DOPE weight ratio (3-25%) is reported, focusing on the role of the metal cation in the formation of the 3D cubic lattice. This result may represent a significant progress toward a design-based approach to drug delivery. PMID- 18399685 TI - In situ GISAXS study of the formation of mesostructured phases within the pores of anodic alumina membranes. AB - The formation and subsequent transformations of mesostructured silica within the confined tubular environment of anodic alumina membrane (AAM) channels [porous alumina membrane (PAM) channels] were investigated for the first time in situ with grazing incidence small-angle X-ray scattering (GISAXS) techniques, in combination with ex situ transmission electron microscopy (TEM) of the same samples. A better understanding of the mesostructure formation mechanism within the confined space of the AAM pores is a direct result of this study. Three different surfactants were used as the structure-directing agents in acid catalyzed silica synthesis solutions. With ionic cetyltrimethylammonium bromide acting as the structure-directing agent, a columnar hexagonal structure with mesopores oriented parallel to the AAM channels was observed to form directly from the beginning of the synthesis. In samples synthesized with the nonionic surfactants Brij 56 and Pluronic P123, a circular hexagonal structure was found to form first; here, the mesopores are aligned around the circumference of the AAM channels. The circular structure subsequently transforms directly into a columnar hexagonal (P123 surfactant), or a mixture of columnar hexagonal and a new curved lamellar phase with lamellae oriented parallel to the walls of the AAM channels (Brij 56 surfactant). These transformations occur after complete solvent evaporation and therefore differ from a simple evaporation-induced phase formation. The existence of a previously postulated lamellar phase could be proven by GISAXS and TEM investigations. PMID- 18399686 TI - Morphological evolution in dewetting polystyrene/polyhedral oligomeric silsesquioxane thin film bilayers. AB - Morphological evolution in dewetting thin film bilayers of polystyrene (PS) and a polyhedral oligomeric silsesquioxane (POSS), trisilanolphenyl-POSS (TPP), was studied as a function of annealing temperature and annealing time. The results demonstrate unique dewetting morphologies in PS/TPP bilayers at elevated temperatures that are significantly different from those typically observed in dewetting polymer/polymer bilayers. During temperature ramp studies by optical microscopy (OM) in the reflection mode, PS/TPP bilayers form cracks with a weak optical contrast at approximately 130 degrees C. The crack formation is attributed to tensile stresses within the upper TPP layer. The weak optical contrast of the cracks observed in the bilayers for annealing temperatures below approximately 160 degrees C is consistent with the cracking and dewetting of only the upper TPP layer from the underlying PS layer. The optical contrast of the morphological features is significantly enhanced at annealing temperatures of >160 degrees C. This observation suggests dewetting of both the upper TPP and the lower PS layers that results in the exposure of the silicon substrate. Upon annealing the PS/TPP bilayers at 200 degrees C in a temperature jump experiment, the upper TPP layer undergoes instantaneous cracking as observed by OM. These cracks in the upper TPP layer serve as nucleation sites for rapid dewetting and aggregation of the TPP layer, as revealed by OM and atomic force microscopy (AFM). X-ray photoelectron spectroscopy (XPS) results indicated that dewetting of the lower PS layer ensued for annealing times >5 min and progressed up to 90 min. For annealing times >90 min, OM, AFM, and XPS results revealed complete dewetting of both the layers with the formation of TPP encapsulated PS droplets. PMID- 18399687 TI - Phase separation in poly(tert-butyl acrylate)/polyhedral oligomeric silsesquioxane (POSS) thin film blends. AB - Phase separation in thin film blends of poly(tert-butyl acrylate) (PtBA) and a polyhedral oligomeric silsesquioxane (POSS), trisilanolphenyl-POSS (TPP), is studied as functions of annealing temperature and time, using reflected light optical microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy. The results demonstrate that the PtBA/TPP blend system confined to thin films ( approximately 90 nm) exhibits lower critical solution temperature (LCST) behavior with a critical temperature of approximately 70 degrees C and a critical composition of 60 wt % PtBA with insignificant dewetting at the phase boundary. Off-critical spinodal behavior is observed for 58 and 62 wt % PtBA blend films. Phase separation by nucleation and growth is observed for all compositions outside the window between 58 and 62 wt % PtBA. The temporal evolution of spinodal decomposition in 60 wt % PtBA blend films is explored at annealing temperatures of 75, 85, 95, and 105 degrees C. The morphological evolution in 60 wt % PtBA blend films is similar for all experimental temperatures (75, 85, 95, and 105 degrees C) with the expected shorter time scales for phase evolution at higher annealing temperatures. Fast Fourier transforms of optical micrographs reveal that these blend films immediately undergo phase separation by spinodal decomposition during temperature jump experiments. Power law scaling for the characteristic wavevector with time (q approximately t(n) with n approximately -1/4 to -1/3) for domain growth during the early stages of phase separation yields to domain pinning at the later stages for 60 wt % PtBA blend films annealed at 75, 85, and 95 degrees C. In contrast, domain growth is pinned over the entire experimental time scale for 60 wt % PtBA blend films annealed at 105 degrees C. PMID- 18399688 TI - Fabrication, structural characterization, and applications of langmuir and langmuir-blodgett films of a poly(azo)urethane. AB - The synthesis of a poly(azo)urethane by fixing CO(2) in bis-epoxide followed by a polymerization reaction with an azodiamine is presented. Since isocyanate is not used in the process, it is termed "clean method" and the polymers obtained are named "NIPUs" (non-isocyanate polyurethanes). Langmuir films were formed at the air-water interface and were characterized by surface pressure vs mean molecular area per mer unit (Pi-A) isotherms. The Langmuir monolayers were further studied by running stability tests and cycles of compression/expansion (possible hysteresis) and by varying the compression speed of the monolayer formation, the subphase temperature, and the solvents used to prepare the spreading polymer solutions. The Langmuir-Blodgett (LB) technique was used to fabricate ultrathin films of a particular polymer (PAzoU). It is possible to grow homogeneous LB films of up to 15 layers as monitored using UV-vis absorption spectroscopy. Higher number of layers can be deposited when PAzoU is mixed with stearic acid, producing mixed LB films. Fourier transform infrared (FTIR) absorption spectroscopy and Raman scattering showed that the materials do not interact chemically in the mixed LB films. The atomic force microscopy (AFM) and micro Raman technique (optical microscopy coupled to Raman spectrograph) revealed that mixed LB films present a phase separation distinguishable at micrometer or nanometer scale. Finally, mixed and neat LB films were successfully characterized using impedance spectroscopy at different temperatures, a property that may lead to future application as temperature sensors. Principal component analysis (PCA) was used to correlate the data. PMID- 18399689 TI - Thermocapillary motion of a liquid drop on a horizontal solid surface. AB - The motion of drops of decane on horizontal poly(dimethylsiloxane) (PDMS)-coated glass surfaces resulting from a temperature gradient on the surface is studied experimentally, and a theoretical description of the thermocapillary motion of spherical-cap drops on a horizontal solid surface obtained using the lubrication approximation also is presented. The drop size and the applied temperature gradient are varied in the experiments, and the measured velocities of the drops are compared with predictions from the model. The scalings of the velocity with drop size and with the applied temperature gradient are predicted correctly by the theoretical model, even though the actual velocities are smaller than those predicted. The influence of contact angle hysteresis, which leads to a critical drop size below which drops do not move, is found to be minimal. Unlike in previous studies (Chen, J. Z.; Troian, S. M.; Darhuber, A. A.; Wagner, S. J. Appl. Phys. 2005, 97, 014906; Brzoska, J. B.; Brochard-Wyart, F.; Rondelez, F. Langmuir 1993, 9, 2220), this small critical drop size appears to be independent of the applied temperature gradient. Results also are presented on the deformation of the contact lines of the moving drops in the form of an aspect ratio, and correlated with the temperature difference across the footprints of the drops and the capillary number. PMID- 18399690 TI - Quantitative method for determining the lateral strength of bacterial adhesion and application for characterizing adhesion kinetics. AB - A quantitative method for measuring the shear force required to detach individual adhered bacteria using atomic force microscopy (AFM) was developed. By determining the total compression of the cantilever during cell detachment events, a more accurate means of calculating the applied lateral force necessary to remove individual cells was achieved compared to previous methods. In addition, a tunable assay for monitoring the dynamics of Pseudomonas aeruginosa and Staphylococcus aureus adhesion strength was employed. The accumulation of force measurements over time allowed for the characterization of adhesion strength kinetics. P. aeruginosa reinforced its adhesion to the surface at a rate 7-fold faster than for S. aureus; the average adhesion strength of P. aeruginosa was larger than that of S. aureus at corresponding time points. Adhered cells of the same species and strain demonstrated a range of adhesion forces that broadened with time, indicating that the change in adhesion strength does not proceed uniformly. PMID- 18399691 TI - Molecular dynamics simulations of rhodopsin point mutants at the cytoplasmic side of helices 3 and 6. AB - The present work reports on a structural analysis carried out through different computer simulations of a set of rhodopsin mutants with differential functional features in regard to the wild type. Most of these mutants, whose experimental features had previously been reported [Ramon et al. J Biol Chem 282, 14272-14282 (2007)], were designed to perturb a network of electrostatic interactions located at the cytoplasmic sides of transmembrane helices 3 and 6. Geometric and energetic features derived from the detailed analysis of a series of molecular dynamics simulations of the different rhodopsin mutants, involving positions 134(3.49), 247(6.30), and 251(6.34), suggest that the protein structure is sensitive to these mutations through the local changes induced that extend further to the secondary structure of neighboring helices and, ultimately, to the packing of the helical bundle. Overall, the results obtained highlight the complexity of the analyzed network of electrostatic interactions where the effect of each mutation on protein structure can produce rather specific features. PMID- 18399692 TI - Structural complexes of human adenine phosphoribosyltransferase reveal novel features of the APRT catalytic mechanism. AB - Adenine phosphoribosyltransferase (APRT) is an important enzyme component of the purine recycling pathway. Parasitic protozoa of the order Kinetoplastida are unable to synthesize purines de novo and use the salvage pathway for the synthesis of purine bases rendering this biosynthetic pathway an attractive target for antiparasitic drug design. The recombinant human adenine phosphoribosyltransferase (hAPRT) structure was resolved in the presence of AMP in the active site to 1.76 A resolution and with the substrates PRPP and adenine simultaneously bound to the catalytic site to 1.83 A resolution. An additional structure was solved containing one subunit of the dimer in the apo-form to 2.10 A resolution. Comparisons of these three hAPRT structures with other 'type I' PRTases revealed several important features of this class of enzymes. Our data indicate that the flexible loop structure adopts an open conformation before and after binding of both substrates adenine and PRPP. Comparative analyses presented here provide structural evidence to propose the role of Glu104 as the residue that abstracts the proton of adenine N9 atom before its nucleophilic attack on the PRPP anomeric carbon. This work leads to new insights to the understanding of the APRT catalytic mechanism. PMID- 18399693 TI - Accurate aqueous proton dissociation constants calculations for selected angiotensin-converting enzyme inhibitors. AB - Studies that allow computing values of aqueous proton dissociation constants (pKa), gas phase proton affinities, and the free energy of solvation have been performed for six members of angiotensin-I-converting enzyme (ACE) inhibitor family (captopril, enalaprilat, imidaprilat, ramiprilat, perindoprilat, and spiraprilat). Density functional theory (DFT) calculations using PBE1PBE functional on optimized molecular geometries have been carried out to investigate the thermodynamics of gas-phase protonation. The conductor-like polarizable continuum model (CPCM) solvation method at various levels of theory was applied to calculate the free energy of solvation for the ACE inhibitors and their respective anions. The CPCM solvation calculations were performed on both gas phase and solvent-phase optimized structures. The combination of gas-phase and solvation energies according to the thermodynamic cycle enabled us to compute accurate pKa values for the all studied molecules. PMID- 18399694 TI - Study of multiple unfolding trajectories and unfolded states of the protein GB1 under the physical property space. AB - Forty nine molecular dynamics simulations of unfolding trajectories of the segment B1 of streptococcal protein G (GB1) provide a direct demonstration of the diversity of unfolding pathway and give a statistically utmost unfolding pathway under the physical property space. Twelve physical properties of the protein were chosen to construct a 12-dimensional property space. Then the 12-dimensional property space was reduced to a 3-dimensional principle component property space. Under the property space, the multiple unfolding trajectories look like "trees", which have some common characters. The "root of the tree" corresponds to the native state, the "bole" homologizes the partially unfolded conformations, and the "crown" is in correspondence to the unfolded state. These unfolding trajectories can be divided into three types. The first one has the characters of straight "bole" and "crown" corresponding to a fast two-state unfolding pathway of GB1. The second one has the character of "the standstill in the middle tree bole", which may correspond to a three-state unfolding pathway. The third one has the character of "the circuitous bole" corresponding to a slow two-state unfolding pathway. The fast two-state unfolding pathway is a statistically utmost unfolding pathway or preferred pathway of GB1, which occupies 53% of 49 unfolding trajectories. In the property space all the unfolding trajectories construct a thermal unfolding pathway ensemble of GB1. The unfolding pathway ensemble resembles a funnel that is gradually emanative from the native state ensemble to the unfolded state ensemble. In the property space, the thermal unfolded state distribution looks like electronic cloud in quantum mechanics. The unfolded states of the independent unfolding simulation trajectories have substantial overlaps, indicating that the thermal unfolded states are confined by the physical property values, and the number of protein unfolded state are much less than that was believed before. PMID- 18399695 TI - Condensed representations of protein secondary structure sequences and their application. AB - In this paper, we propose a nongraphical representation for protein secondary structures. By counting the frequency of occurrence of all possible four-tuples (i.e., four-letter words) of a protein secondary structure sequence, we construct a set of 3x3 matrices for the corresponding protein secondary structure sequence. Furthermore, the leading eigenvalues of these matrices are computed and considered as invariants for the protein secondary structure sequences. To illustrate the utility of our approach, we apply it to a set of real data to distinguish protein structural classes. The result indicates that it can be used to complement the classification of protein secondary structures. PMID- 18399696 TI - SNP detection for cytochrome P450 alleles by target-assembled tandem oligonucleotide systems based on exciplexes. AB - We report the first use of exciplex-based split-probes for detection of the wild type and *3 mutant alleles of human cytochrome P450 2C9. A tandem 8-mer split DNA oligonucleotide probe system was designed that allows detection of the complementary target DNA sequence. This exciplex-based fluorescence detector system operates by means of a contiguous hybridization of two oligonucleotide exciplex split-probes to a complementary target nucleic acid target. Each probe oligonucleotide is chemically modified at one of its termini by a potential exciplex-forming partner, each of which is fluorescently silent at the wavelength of detection. Under conditions that ensure correct three-dimensional assembly, the chemical moieties on suitable photoexcitation form an exciplex that fluoresces with a large Stokes shift (in this case 130 nm). Preliminary proof-of concept studies used two 8-mer probe oligonucleotides, but in order to give better specificity for genomic applications, probe length was extended to give coverage of 24 bases. Eight pairs of tandem 12-mer oligonucleotide probes spanning the 2C9*3 region were designed and tested to find the best set of probes. Target sequences tested were in the form of (i) synthetic oligonucleotides, (ii) embedded in short PCR products (150 bp), or (iii) inserted into plasmid DNA (approximately 3 Kbp). The exciplex system was able to differentiate wild type and human cytochrome P450 2C9 *3 SNP (1075 A-->C) alleles, based on fluorescence emission spectra and DNA melting curves, indicating promise for future applications in genetic testing and molecular diagnostics. PMID- 18399697 TI - Influence of ionic strength on Hoechst 33258 binding with DNA. AB - The binding of Hoechst 33258 with DNA at various ionic strengths of solution and different ligand concentrations has been investigated. Existence of more than one type of interactions of Hoechst 33258 with DNA has been revealed, which were very sensitive to the ionic strength. Hoechst 33258 doesn't show specificity to AT sequences of DNA at low ionic strength. High affinity binding mode becomes obvious at high ionic strength. The values of binding constants and binding site sizes for revealed strong and weak interactions have been determined. PMID- 18399698 TI - Excessive clustering of third codon position pyrimidines in prokaryotes. AB - The third codon positions are generally thought to be largely neutral, allowing for synonymous mutations. To see how much the third positions are loaded in general we analyzed the sequences of the nucleotides in the third positions. Simple word count analysis revealed excessive clustering of pyrimidines in the third position sequences of prokaryotic mRNA. The clusters have a clear tendency to follow one after another at characteristic distance of 25-30 triplets. Thus, the third codon positions do carry a rather strong message. Possible connection with loop-fold structure of proteins and (cotranslational) protein folding is discussed. PMID- 18399699 TI - Delving deeper into the unexpected correlation between gene expressivity and codon usage bias of Escherichia coli genome. AB - Biased usage of synonymous codons has been elucidated under the perspective of cellular tRNA abundance and more recently by the mRNA secondary structure folding stability of the corresponding genes. Taking advantage of publicly available gene expression data for Escherichia coli, a comprehensive investigation of the three classes of genes having different codon usage biases was performed from the standpoint of tRNA abundance, mRNA secondary structure folding stability, and translational error minimization procedure. We detected the different evolutionary forces for translational and/or transcriptional regulation of highly expressed genes depending upon their codon bias. Additionally, the possible role of mRNA folding stability in maintaining the overall high expressivity of the set of lowly biased genes has been articulated. These novel findings certainly strengthen the understanding of the codon usage bias in the Escherichia coli genome. PMID- 18399700 TI - 2-aminopurine fluorescence: discrimination between specific and unspecific ligand binding to the kissing-loop dimer of the HIV-1 RNA. AB - The fluorescent 2-aminopurine probe (2-AP) incorporated into the loop of 23-mer RNA hairpin of HIV-1 genome dimerization initiation site (DIS) was used for discrimination of specific and unspecific binding of paromomycin and spermine to the kissing loop dimer (KD) formed in solution. While both ligands stabilized the KD RNA structure, only paromomycin binding resulted in significant increase of 2 AP fluorescence. These observations suggest that the 2-AP fluorescent RNA construct might be useful for selecting ligands specifically binding the HIV-1 kissing loop RNA dimer. PMID- 18399701 TI - RNA2D3D: a program for generating, viewing, and comparing 3-dimensional models of RNA. AB - Using primary and secondary structure information of an RNA molecule, the program RNA2D3D automatically and rapidly produces a first-order approximation of a 3 dimensional conformation consistent with this information. Applicable to structures of arbitrary branching complexity and pseudoknot content, it features efficient interactive graphical editing for the removal of any overlaps introduced by the initial generating procedure and for making conformational changes favorable to targeted features and subsequent refinement. With emphasis on fast exploration of alternative 3D conformations, one may interactively add or delete base-pairs, adjacent stems can be coaxially stacked or unstacked, single strands can be shaped to accommodate special constraints, and arbitrary subsets can be defined and manipulated as rigid bodies. Compaction, whereby base stacking within stems is optimally extended into connecting single strands, is also available as a means of strategically making the structures more compact and revealing folding motifs. Subsequent refinement of the first-order approximation, of modifications, and for the imposing of tertiary constraints is assisted with standard energy refinement techniques. Previously determined coordinates for any part of the molecule are readily incorporated, and any part of the modeled structure can be output as a PDB or XYZ file. Illustrative applications in the areas of ribozymes, viral kissing loops, viral internal ribosome entry sites, and nanobiology are presented. PMID- 18399702 TI - Fuzzy kernel clustering of RNA secondary structure ensemble using a novel similarity metric. AB - Measuring the (dis)similarity between RNA secondary structures is critical for the study of RNA secondary structures and has implications to RNA functional characterization. Although a number of methods have been developed for comparing RNA structural similarities, their applications have been limited by the complexity of the required computation. In this paper, we present a novel method for comparing the similarity of RNA secondary structures generated from the same RNA sequence, i.e., a secondary structure ensemble, using a matrix representation of the RNA structures. Relevant features of the RNA secondary structures can be easily extracted through singular value decomposition (SVD) of the representing matrices. We have mapped the feature vectors of the singular values to a kernel space, where (dis)similarities among the mapped feature vectors become more evident, making clustering of RNA secondary structures easier to handle. The pair wise comparison of RNA structures is achieved through computing the distance between the singular value vectors in the kernel space. We have applied a fuzzy kernel clustering method, using this similarity metric, to cluster the RNA secondary structure ensembles. Our application results suggest that our fuzzy kernel clustering method is highly promising for classifications of RNA structure ensembles, because of its low computational complexity and high clustering accuracy. PMID- 18399703 TI - Quantum molecular modeling of glycyl-adenylate. AB - Glycyl-adenylate is the simplest model for understanding the central role played by aminoacyl-adenylates in the protein synthesis, and may be also in the prebiotic evolution. Optimized conformation of glycyl-adenylate (GlyAMP), in vacuum, was determined using ab initio RHF/6-31Gdp computations. GlyAMP in vacuum (form corresponding to neutral pH conditions) possesses a specific conformation of the glycyl-phosphate stem with a double seven membered ring that results from the electrostatic interaction between the anionic phosphate and the cationic ammonium. This structure is explained on the basis of the theoretical infrared vibrational spectrum and discussed with regard to those of glycyl-adenylate (GAP1550 and GAP2550 forms) in the active pockets of glycyl-tRNA synthetase. The electrostatic properties of these different conformations are presented, and the influence of the charged residues (anionic GLU188, GLU239, GLU359, and cationic ARG220) and Mg++, in interaction with GAP1550 are discussed, taking into account the computation of the electronic chemical potential, the electron affinity and the electrophilic index. PMID- 18399704 TI - Quantum chemical studies of structures and binding in noncanonical RNA base pairs: the trans Watson-Crick:Watson-Crick family. AB - The trans Watson-Crick/Watson-Crick family of base pairs represent a geometric class that play important structural and possible functional roles in the ribosome, tRNA, and other functional RNA molecules. They nucleate base triplets and quartets, participate as loop closing terminal base pairs in hair pin motifs and are also responsible for several tertiary interactions that enable sequentially distant regions to interact with each other in RNA molecules. Eleven representative examples spanning nine systems belonging to this geometric family of RNA base pairs, having widely different occurrence statistics in the PDB database, were studied at the HF/6-31G (d, p) level using Morokuma decomposition, Atoms in Molecules as well as Natural Bond Orbital methods in the optimized gas phase geometries and in their crystal structure geometries, respectively. The BSSE and deformation energy corrected interaction energy values for the optimized geometries are compared with the corresponding values in the crystal geometries of the base pairs. For non protonated base pairs in their optimized geometry, these values ranged from -8.19 kcal/mol to -21.84 kcal/mol and compared favorably with those of canonical base pairs. The interaction energies of these base pairs, in their respective crystal geometries, were, however, lesser to varying extents and in one case, that of A:A W:W trans, it was actually found to be positive. The variation in RMSD between the two geometries was also large and ranged from 0.32 2.19 A. Our analysis shows that the hydrogen bonding characteristics and interaction energies obtained, correlated with the nature and type of hydrogen bonds between base pairs; but the occurrence frequencies, interaction energies, and geometric variabilities were conspicuous by the absence of any apparent correlation. Instead, the nature of local interaction energy hyperspace of different base pairs as inferred from the degree of their respective geometric variability could be correlated with the identities of free and bound hydrogen bond donor/acceptor groups present in interacting bases in conjunction with their tertiary and neighboring group interaction potentials in the global context. It also suggests that the concept of isostericity alone may not always determine covariation potentials for base pairs, particularly for those which may be important for RNA dynamics. These considerations are more important than the absolute values of the interaction energies in their respective optimized geometries in rationalizing their occurrences in functional RNAs. They highlight the importance of revising some of the existing DNA based structure analysis approaches and may have significant implications for RNA structure and dynamics, especially in the context of structure prediction algorithms. PMID- 18399705 TI - Studies on tautomeric forms of Guanine-Cytosine base pairs of nucleic acids and their interactions with water molecules. AB - The relative stabilities of Guanine-Cytosine (G-C) DNA bare base pairs, its tautomeric forms and microhydrated base pairs are theoretically investigated with a focus on the keto-enol tautomerism as well as on the cis-trans isomerism using ab initio and density functional theory methods. The stabilities of the G-C bare base pairs, its tautomeric forms and microhydrated base pairs were affected by various factors including keto-enol tautomerization, cis-trans enol isomerization, and steric hindrance between the base pair and water molecules. The Atoms in Molecules theory (AIM) is employed to investigate H-bonding patterns both in bare and microhydrated base pairs. From the above topological results, an excellent linear correlation is shown between electron density [rho(r)], and its Laplacian [V2rho(r)] at the bond critical points. NBO analysis has been carried out to study the charge transfer between proton acceptor to the antibonding orbital of the X-H bond both in bare and microhydrated base pairs. PMID- 18399706 TI - Internet addiction: definition, assessment, epidemiology and clinical management. AB - Internet addiction is characterized by excessive or poorly controlled preoccupations, urges or behaviours regarding computer use and internet access that lead to impairment or distress. The condition has attracted increasing attention in the popular media and among researchers, and this attention has paralleled the growth in computer (and Internet) access. Prevalence estimates vary widely, although a recent random telephone survey of the general US population reported an estimate of 0.3-0.7%. The disorder occurs worldwide, but mainly in countries where computer access and technology are widespread. Clinical samples and a majority of relevant surveys report a male preponderance. Onset is reported to occur in the late 20s or early 30s age group, and there is often a lag of a decade or more from initial to problematic computer usage. Internet addiction has been associated with dimensionally measured depression and indicators of social isolation. Psychiatric co-morbidity is common, particularly mood, anxiety, impulse control and substance use disorders. Aetiology is unknown, but probably involves psychological, neurobiological and cultural factors. There are no evidence-based treatments for internet addiction. Cognitive behavioural approaches may be helpful. There is no proven role for psychotropic medication. Marital and family therapy may help in selected cases, and online self-help books and tapes are available. Lastly, a self-imposed ban on computer use and Internet access may be necessary in some cases. PMID- 18399708 TI - Burden of bipolar depression: impact of disorder and medications on quality of life. AB - Bipolar disorder is a complex, chronic psychiatric condition characterized by recurring episodes of depressive illness and mania or hypomania. Although the manic or hypomanic episodes define the disorder, recent research has shown that depressive symptoms predominate over manic symptoms in the majority of patients, and that bipolar depression accounts for much of the significant morbidity and mortality associated with bipolar disorder. Given these findings, there has been a recent upsurge of interest in furthering our understanding of the burden of depression in bipolar disorder. At the same time, increasing scientific attention is now being paid to expanding the measurement of outcome in bipolar disorder to encompass broader indicators of response, one of which is the assessment of quality of life (QOL). In this review, we provide a summary of the current knowledge about QOL in the depressive phase of bipolar disorder, and the effects of pharmacological treatment interventions for bipolar disorder upon QOL. It appears that QOL is poorer in bipolar disorder than in other mood disorders and anxiety disorders, but that schizophrenia might compromise QOL more severely than bipolar disorder. Existing data also suggest that, for patients with bipolar disorder, QOL is negatively associated with depression, both as a cross-sectional mood state and perhaps also as a feature of the patient's course. Despite its clinical and public health importance, bipolar depression has only recently started to receive the attention it warrants in clinical trials, and many important questions about its optimal pharmacological management remain to be answered. There is also a paucity of information about the impact of pharmacological interventions on QOL in bipolar depression. To our knowledge, only two clinical trials to date have specifically examined the impact of medications on QOL in patients with bipolar depression. A small number of other studies have examined the effects of depressive symptoms on QOL in patients who are experiencing manic or mixed episodes. Nonetheless, QOL appears to be a meaningful and important indicator of outcome and recovery in this patient population, and one that warrants further scientific interest and energy. PMID- 18399709 TI - Antiparkinsonian medication and pathological gambling. AB - Parkinson's disease is a common condition, usually treated by dopaminergic agents, both ergot and non-ergot. Many behavioural abnormalities are associated with such usage, including impulse control disorders (ICDs), dopamine dysregulation syndrome and 'punding'. Pathological gambling, a form of ICD, comprises persistent and maladaptive gambling of various types that disrupts personal, family or occupational activity. Pathological gambling may be associated with other abnormal actions such as pathological shopping, hoarding and hypersexuality. The incidence varies widely from study to study but may be up to 7% of users of dopaminergic agents. Recognition of this problem has led drug regulatory agencies to add precautions concerning pathological gambling to official drug information for the entire class of antiparkinsonian medications. The literature is not entirely consistent and opinions differ greatly, but pramipexole (a dopamine D2 and D3 agonist), and perhaps ropinirole (also a D2/D3 agonist), may be especially likely to be associated with pathological gambling, although the precise nature of the relationship is unclear. Treatment involves reducing the dose of the medication or switching to another medication; unfortunately, the Parkinson's disease may worsen. The mechanism of this adverse effect is believed to be excessive dopaminergic stimulation but probably not specifically involving D3 receptors. A parallel to addictive behaviour with stimulant drugs has been noted. PMID- 18399707 TI - Aripiprazole in the treatment of depressive and anxiety disorders: a review of current evidence. AB - Despite the availability of different classes of drugs for the treatment of depressive and anxiety disorders, there are a number of clinically significant unmet needs, such as a high prevalence of treatment resistance, partial response, subsyndromal symptomatology, recurrence and relapse. With the approval of atypical antipsychotics, which are associated with a lower adverse effect burden than typical antipsychotics, consideration of their off-label use for the treatment of affective disorders and various other psychiatric disorders has become a viable option. However, consideration should be given to the US FDA black box warning indicating that atypical antipsychotics may increase mortality risk, particularly in the elderly population with dementia-related psychosis. There has been much conjecture about the utility of these atypical drugs to facilitate traditional antidepressant therapy, either in combination (from the initiation of therapy) or as adjunctive therapy (in the case of partial/incomplete response). Nevertheless, at present, available evidence from randomized, placebo-controlled trials is sparse, and a formal risk/benefit assessment of the use of these agents in a nonpsychotic patient population is not yet possible. As a representative agent from the atypical antipsychotic class with a novel mechanism of action and a relatively low adverse effect burden, aripiprazole represents an interesting potential treatment for depressive and anxiety disorders. In this review, we focus on the rationale for the use of aripiprazole in these disorders. Preclinical data suggests that aripiprazole has a number of possible mechanisms of action that may be important in the treatment of depressive and anxiety disorders. Such mechanisms include aripiprazole action at serotonin (5-HT) receptors as a 5-HT1A partial receptor agonist, a 5-HT2C partial receptor agonist and a 5-HT2A receptor antagonist. Aripiprazole also acts as a dopamine D2 partial receptor agonist, and has a possible action at adrenergic receptors. Furthermore, aripiprazole may have possible neuroprotective effects. Clinical studies demonstrate that aripiprazole may be useful in the treatment of bipolar depression, major depressive disorder, treatment-resistant depression and possibly anxiety disorders. Clinical data also suggest that aripiprazole may have a lower adverse effect burden than the other atypical drugs. Future research may confirm the potential utility of aripiprazole in the treatment of depressive and anxiety disorders. PMID- 18399711 TI - Clinical pharmacokinetics and pharmacodynamics of the oral direct thrombin inhibitor dabigatran etexilate. AB - The direct thrombin inhibitor dabigatran etexilate is currently in phase III of development for the prophylaxis and treatment of thromboembolic disorders, with three trials completed in primary venous thromboembolism (VTE) prevention. Dabigatran etexilate is an orally administered prodrug, which is rapidly absorbed and converted to the active form, dabigatran. Dabigatran has been shown to specifically and reversibly inhibit thrombin, the key enzyme in the coagulation cascade. Studies in healthy volunteers and in patients undergoing orthopaedic surgery have indicated that dabigatran has a predictable pharmacokinetic/pharmacodynamic profile, allowing for a fixed-dose regimen. Peak plasma concentrations of dabigatran are reached approximately 2 hours after oral administration in healthy volunteers, with no unexpected accumulation of drug concentrations upon multiple dosing. Excretion is predominantly via the renal route as unchanged drug. Dabigatran is not metabolized by cytochrome P450 isoenzymes. The small differences in dabigatran pharmacokinetics associated with age and gender are attributed to variations in renal function. Additional studies have shown that the pharmacokinetic/pharmacodynamic profile of dabigatran is consistent across a range of patient populations, with no effect of moderate hepatic impairment being observed. Drug-drug interactions are not observed with concomitant administration of atorvastatin, diclofenac or digoxin. The pharmacodynamic profile of dabigatran demonstrates effective anticoagulation combined with a low risk of bleeding. Further phase III studies are ongoing, including acute VTE treatment and stroke prevention in atrial fibrillation; the results obtained so far show that dabigatran etexilate is well tolerated and effective in the treatment and prevention of thromboembolic events. PMID- 18399710 TI - An update on the pharmacological management of post-herpetic neuralgia and painful diabetic neuropathy. AB - Neuropathic pain is a persistent pain condition that develops secondary to nerve injury. The two most common types of peripheral neuropathic pain are post herpetic neuralgia (PHN) and painful diabetic neuropathy (PDN). Amitriptyline, nortriptyline, desipramine and imipramine are TCAs that have been shown to be effective for the symptomatic relief of PHN and PDN. Serotonin noradrenaline reuptake inhibitors (SNRIs) such as venlafaxine and duloxetine have been shown to be very promising for the treatment of PDN with fewer adverse effects than TCAs. Selective serotonin reuptake inhibitors (SSRIs) were shown in a number of studies to have some efficacy in relieving PDN-related pain, yet other studies of the SSRIs have demonstrated conflicting outcomes. Most of the older antiepileptic studies were performed in patients with PDN; consequently, little is known about the efficacy of these drugs in patients with PHN. Carbamazepine, phenytoin and valproic acid were shown to be effective in ameliorating PDN-related pain. Other antiepileptic agents, including lamotrigine, oxcarbazepine and topiramate, have demonstrated some beneficial effects for the treatment of PDN, although they were also found to be ineffective in some PDN studies. alpha2delta Ligands such as gabapentin and pregabalin have been proven to be effective for the treatment of PHN and PDN in a number of large placebo-controlled trials. These drugs are useful not only in relieving pain but also in improving quality of life. Although the use of opioids for the treatment of neuropathic pain is controversial, a number of studies support the efficacy and safety of opioids in the treatment of neuropathic pain. Of these, oxycodone and tramadol have been shown to be superior to placebo for the treatment of PHN and PDN. A number of small studies have shown that dextromethorphan was effective in patients with PDN but not in patients with PHN. Topical agents such as lidocaine 5% patches and topical capsaicin are useful in ameliorating pain in patients with PHN but these agents are unsatisfactory for use as a sole agent. Although a number of drug treatments are available for the symptomatic relief of neuropathic pain symptoms, these agents do not provide satisfactory relief in all patients. For these patients, other treatment alternatives such as combination drug therapy that produces pain relief via distinctly different mechanisms may be successful. The purpose of this review is to compare the efficacy and limitations of currently available pharmacological treatments for the symptomatic relief of PHN and PDN, and to discuss the potential of combination therapy in PHN and PDN. PMID- 18399712 TI - Free drug metabolic clearance in elderly people. AB - The question of whether metabolic drug clearance is decreased in elderly people has been the subject of considerable debate and is very important because clearance is a determinant of dosing. Drug clearance has been shown to be consistently impaired for flow-limited (high-clearance) drugs, but there have been conflicting results for capacity-limited (low-clearance) drugs. A limitation of the studies of capacity-limited drugs is that most have estimated clearance based on total drug concentrations (protein-bound plus free). Total drug clearance reflects both the intrinsic clearance of free drug and the extent of protein binding. Total clearance is a valid measure for capacity-limited drugs with low protein binding and appears to be consistently impaired in elderly subjects. For phenazone [antipyrine] (fraction unbound [f(u)] >0.9), seven studies have demonstrated statistical reductions in clearance of 20-52%. For theophylline (f(u) 0.6), five studies have demonstrated reductions in clearance of 22-35%. For paracetamol [acetaminophen] (f(u) 0.8), the clearance of which has been quoted as unchanged, four studies have demonstrated reductions in clearance of 19-35%. For highly protein-bound drugs, total clearance is not the appropriate parameter. Free drug clearance is more appropriate since it is independent of changes in protein binding. The literature was reviewed to test the hypothesis that in elderly people, capacity-limited drugs with high protein binding will show decreased free clearance even in the absence of a decrease in total clearance. For these drugs, data for free drug clearance based on measurement of actual free drug concentrations are limited, but suggest that the intrinsic metabolic clearance is impaired in elderly subjects. Four studies of naproxen (f(u) <0.01) have shown reduced free drug clearance of 50% or more. Two studies of valproic acid (f(u) 0.1-0.2) have shown reduced free clearance of 39% and 65%. Two studies of ibuprofen (f(u) <0.01) have shown reduced free clearance of S ibuprofen of 21% and 28%. There is some indirect evidence for reduced clearance of the highly protein-bound drugs oxaprozin, temazepam, lorazepam, diazepam, phenytoin and warfarin, although studies measuring free concentrations are lacking. Together, the above studies support the hypothesis that the intrinsic metabolic drug clearance is impaired in elderly subjects, in the order of 20-60%, and that this effect is masked if highly protein-bound drugs are assessed using total drug clearance. If the findings are confirmed in future well-designed studies of free drug clearance, there are profound and beneficial implications for dosing of drugs in elderly people. Lower doses are likely to achieve appropriate concentrations, allowing full efficacy but decreased dose-related adverse effects. PMID- 18399713 TI - Determinants of steady-state torasemide pharmacokinetics: impact of pharmacogenetic factors, gender and angiotensin II receptor blockers. AB - BACKGROUND: Torasemide is frequently used for the treatment of hypertension and heart failure. However, the determinants of torasemide pharmacokinetics in patients during steady-state conditions are largely unknown. We therefore explored the impact of genetic polymorphisms of cytochrome P450 (CYP) 2C9 (CYP2C9) and organic anion transporting polypeptide (OATP) 1B1 (SLCO1B1), gender, and the effects of losartan and irbesartan comedication on the interindividual variability of steady-state pharmacokinetics of torasemide. PATIENTS AND METHODS: Twenty-four patients receiving stable medication with torasemide 10 mg once daily and with an indication for additional angiotensin II receptor blocker (ARB) treatment to control hypertension or to treat heart failure were selected. Blood samples were taken before torasemide administration and 0.5, 1, 2, 4, 8, 12 and 24 hours after administration. After this first study period, patients received either irbesartan 150 mg (five female and seven male patients aged 69+/-8 years) or losartan 100 mg (two female and ten male patients aged 61+/-8 years) once daily. After 3 days of ARB medication, eight blood samples were again collected at the timepoints indicated above. The patients' long-term medications, which did not include known CYP2C9 inhibitors, were maintained at a constant dose during the study. All patients were genotyped for CYP2C9 (*1/*1 [n=15]; *1/*2 [n = 4]; *1/*3 [n=5]) as well as for SLCO1B1 (c.521TT [n=13]; c.521TC [n=11]). RESULTS: Factorial ANOVA revealed an independent impact of the CYP2C9 genotype (dose normalized area under the plasma concentration-time curve during the 24-hour dosing interval at steady state [AUC(24,ss)/D]: *1/*1 375.5+/-151.4 microg x h/L/mg vs *1/*3 548.5+/-271.6 microg x h/L/mg, p=0.001), the SLCO1B1 genotype (AUC(24,ss)/D: TT 352.3+/-114 microg x h/L/mg vs TC 487.6+/-218.4 microg x h/L/mg, p<0.05) and gender (AUC(24,ss)/D: males 359.5+/-72.2 microg x h/L/mg vs females 547.3+/-284 microg x h/L/mg, p<0.01) on disposition of torasemide. Coadministration of irbesartan caused a 13% increase in the AUC(24,ss)/D of torasemide (p=0.002), whereas losartan had no effect. CONCLUSION: This study shows that the CYP2C9*3 and SLCO1B1 c.521TC genotype and female gender are significant and independent predictors of the pharmacokinetics of torasemide. Coadministration of irbesartan yields moderate but significant increases in the torasemide plasma concentration and elimination half-life. PMID- 18399714 TI - Retrospective population pharmacokinetic analysis of levetiracetam in children and adolescents with epilepsy: dosing recommendations. AB - OBJECTIVE: To characterize levetiracetam pharmacokinetics, identify significant covariate relationships and identify doses in children that achieve blood concentrations similar to those observed in adults. METHODS: Nonlinear mixed effects modelling was used to analyse pooled data collected from 228 children with epilepsy aged 3 months to 18 years in five trials of adjunctive levetiracetam therapy. Simulations were used to identify dosing regimens achieving levetiracetam steady-state peak and trough plasma concentrations similar to those attained in adults receiving the recommended starting dose for adjunctive therapy (500 mg twice daily). The covariates considered for inclusion in the base model were age, bodyweight, gender, race, body surface area (BSA), body mass index (BMI), creatinine clearance (CL(CR)), levetiracetam dose, concomitant antiepileptic drug (AED) by category (neutral, enzyme inducer, inhibitor, combination of inducer and inhibitor), and benzodiazepines. RESULTS: A one-compartment model with first-order absorption and elimination best characterized the data. The following significant covariates were identified: (i) age on the absorption rate constant (k(a)); (ii) bodyweight, dose, CL(CR) and concomitant enzyme-inducing AED on plasma oral clearance (CL/F); and (iii) bodyweight on the apparent volume of distribution after oral administration (V(d)/F). The main explanatory covariates were age on k(a), bodyweight on CL/F and V(d)/F, and enzyme-inducing AED on CL/F, of which bodyweight was the most influential covariate. Dosing can be carried out with either 10 mg/kg of oral solution twice daily in children weighing <50 kg and a 500-mg tablet twice daily in those weighing >50 kg or, when patients favour a solid formulation, 10 mg/kg of oral solution twice daily in children weighing <20 kg, a 250-mg tablet twice daily in those weighing 20-40 kg, and a 500-mg tablet twice daily in those weighing >40 kg. All of these doses achieved steady-state peak and trough plasma concentrations similar to those observed in adults following the recommended starting dose for adjunctive therapy (500 mg twice daily). CONCLUSIONS: The most influential covariate of levetiracetam pharmacokinetics in children is bodyweight. A starting dose of levetiracetam 10 mg/kg twice daily ensures the same exposure in children as does 500 mg twice daily in adults. PMID- 18399715 TI - Effect of coadministered ketoconazole, a strong cytochrome P450 3A4 enzyme inhibitor, on the pharmacokinetics of ciclesonide and its active metabolite desisobutyryl-ciclesonide. AB - BACKGROUND AND OBJECTIVES: Cytochrome P450 (CYP) 3A4 isoenzyme has been identified in vitro as the key enzyme to metabolize desisobutyryl-ciclesonide (des-CIC), the pharmacologically active metabolite of the inhaled corticosteroid ciclesonide. This pharmacokinetic drug-drug interaction study was conducted to confirm this major metabolic pathway in vivo by using the strong CYP3A4 inhibitor ketoconazole, and to assess the effect of ketoconazole on the pharmacokinetics of ciclesonide and des-CIC. METHODS: Fourteen healthy adults participated in this open-label, nonrandomized, fixed sequence, two-period, repeated-dose pharmacokinetic study. During the first 7-day treatment period, the subjects orally inhaled ciclesonide 320 microg once daily. During the second 7-day treatment period, the subjects continued with the same dose of orally inhaled ciclesonide and concomitantly received oral ketoconazole 400 mg once daily. Pharmacokinetic profiles for ciclesonide and des-CIC were obtained on day 7 of each study period. RESULTS: For the parent compound, ciclesonide, no changes in the pharmacokinetic parameter estimates--the area under the serum concentration time curve during the dosage interval (AUC(tau)), maximum serum concentration (C(max)) and time to reach the C(max)--were observed. In contrast, the AUC(tau) and C(max) of des-CIC increased approximately 3.5-fold and 2-fold under the influence of the CYP3A4 inhibitor ketoconazole. CONCLUSIONS: The CYP3A4 pathway is the major pathway for biotransformation of the active metabolite of ciclesonide in humans. While elimination of des-CIC was reduced by strong CYP3A4 inhibitor coadministration in vivo, activation of the parent compound ciclesonide to des-CIC was not affected. Dose adjustment is not necessary when ciclesonide needs to be coadministered with ketoconazole, because the potency of an inhaled corticosteroid is mediated by topical concentrations in the lung and because ciclesonide has a very low potential to produce systemic adverse effects. PMID- 18399716 TI - Long-term interleukin-6 levels and subsequent risk of coronary heart disease: two new prospective studies and a systematic review. AB - BACKGROUND: The relevance to coronary heart disease (CHD) of cytokines that govern inflammatory cascades, such as interleukin-6 (IL-6), may be underestimated because such mediators are short acting and prone to fluctuations. We evaluated associations of long-term circulating IL-6 levels with CHD risk (defined as nonfatal myocardial infarction [MI] or fatal CHD) in two population-based cohorts, involving serial measurements to enable correction for within-person variability. We updated a systematic review to put the new findings in context. METHODS AND FINDINGS: Measurements were made in samples obtained at baseline from 2,138 patients who had a first-ever nonfatal MI or died of CHD during follow-up, and from 4,267 controls in two cohorts comprising 24,230 participants. Correction for within-person variability was made using data from repeat measurements taken several years apart in several hundred participants. The year-to-year variability of IL-6 values within individuals was relatively high (regression dilution ratios of 0.41, 95% confidence interval [CI] 0.28-0.53, over 4 y, and 0.35, 95% CI 0.23 0.48, over 12 y). Ignoring this variability, we found an odds ratio for CHD, adjusted for several established risk factors, of 1.46 (95% CI 1.29-1.65) per 2 standard deviation (SD) increase of baseline IL-6 values, similar to that for baseline C-reactive protein. After correction for within-person variability, the odds ratio for CHD was 2.14 (95% CI 1.45-3.15) with long-term average ("usual") IL-6, similar to those for some established risk factors. Increasing IL-6 levels were associated with progressively increasing CHD risk. An updated systematic review of electronic databases and other sources identified 15 relevant previous population-based prospective studies of IL-6 and clinical coronary outcomes (i.e., MI or coronary death). Including the two current studies, the 17 available prospective studies gave a combined odds ratio of 1.61 (95% CI 1.42-1.83) per 2 SD increase in baseline IL-6 (corresponding to an odds ratio of 3.34 [95% CI 2.45 4.56] per 2 SD increase in usual [long-term average] IL-6 levels). CONCLUSIONS: Long-term IL-6 levels are associated with CHD risk about as strongly as are some major established risk factors, but causality remains uncertain. These findings highlight the potential relevance of IL-6-mediated pathways to CHD. PMID- 18399717 TI - Quantifying the importance of interleukin-6 for coronary heart disease. PMID- 18399718 TI - Modelling within-host spatiotemporal dynamics of invasive bacterial disease. AB - Mechanistic determinants of bacterial growth, death, and spread within mammalian hosts cannot be fully resolved studying a single bacterial population. They are also currently poorly understood. Here, we report on the application of sophisticated experimental approaches to map spatiotemporal population dynamics of bacteria during an infection. We analyzed heterogeneous traits of simultaneous infections with tagged Salmonella enterica populations (wild-type isogenic tagged strains [WITS]) in wild-type and gene-targeted mice. WITS are phenotypically identical but can be distinguished and enumerated by quantitative PCR, making it possible, using probabilistic models, to estimate bacterial death rate based on the disappearance of strains through time. This multidisciplinary approach allowed us to establish the timing, relative occurrence, and immune control of key infection parameters in a true host-pathogen combination. Our analyses support a model in which shortly after infection, concomitant death and rapid bacterial replication lead to the establishment of independent bacterial subpopulations in different organs, a process controlled by host antimicrobial mechanisms. Later, decreased microbial mortality leads to an exponential increase in the number of bacteria that spread locally, with subsequent mixing of bacteria between organs via bacteraemia and further stochastic selection. This approach provides us with an unprecedented outlook on the pathogenesis of S. enterica infections, illustrating the complex spatial and stochastic effects that drive an infectious disease. The application of the novel method that we present in appropriate and diverse host-pathogen combinations, together with modelling of the data that result, will facilitate a comprehensive view of the spatial and stochastic nature of within-host dynamics. PMID- 18399719 TI - Rapid encoding and perception of novel odors in the rat. AB - To gain insight into which parameters of neural activity are important in shaping the perception of odors, we combined a behavioral measure of odor perception with optical imaging of odor representations at the level of receptor neuron input to the rat olfactory bulb. Instead of the typical test of an animal's ability to discriminate two familiar odorants by exhibiting an operant response, we used a spontaneously expressed response to a novel odorant-exploratory sniffing-as a measure of odor perception. This assay allowed us to measure the speed with which rats perform spontaneous odor discriminations. With this paradigm, rats discriminated and began responding to a novel odorant in as little as 140 ms. This time is comparable to that measured in earlier studies using operant behavioral readouts after extensive training. In a subset of these trials, we simultaneously imaged receptor neuron input to the dorsal olfactory bulb with near-millisecond temporal resolution as the animal sampled and then responded to the novel odorant. The imaging data revealed that the bulk of the discrimination time can be attributed to the peripheral events underlying odorant detection: receptor input arrives at the olfactory bulb 100-150 ms after inhalation begins, leaving only 50-100 ms for central processing and response initiation. In most trials, odor discrimination had occurred even before the initial barrage of receptor neuron firing had ceased and before spatial maps of activity across glomeruli had fully developed. These results suggest a coding strategy in which the earliest-activated glomeruli play a major role in the initial perception of odor quality, and place constraints on coding and processing schemes based on simple changes in spike rate. PMID- 18399720 TI - Control of apoptosis by asymmetric cell division. AB - Asymmetric cell division and apoptosis (programmed cell death) are two fundamental processes that are important for the development and function of multicellular organisms. We have found that the processes of asymmetric cell division and apoptosis can be functionally linked. Specifically, we show that asymmetric cell division in the nematode Caenorhabditis elegans is mediated by a pathway involving three genes, dnj-11 MIDA1, ces-2 HLF, and ces-1 Snail, that directly control the enzymatic machinery responsible for apoptosis. Interestingly, the MIDA1-like protein GlsA of the alga Volvox carteri, as well as the Snail-related proteins Snail, Escargot, and Worniu of Drosophila melanogaster, have previously been implicated in asymmetric cell division. Therefore, C. elegans dnj-11 MIDA1, ces-2 HLF, and ces-1 Snail may be components of a pathway involved in asymmetric cell division that is conserved throughout the plant and animal kingdoms. Furthermore, based on our results, we propose that this pathway directly controls the apoptotic fate in C. elegans, and possibly other animals as well. PMID- 18399721 TI - Darwinian evolution on a chip. AB - Computer control of Darwinian evolution has been demonstrated by propagating a population of RNA enzymes in a microfluidic device. The RNA population was challenged to catalyze the ligation of an oligonucleotide substrate under conditions of progressively lower substrate concentrations. A microchip-based serial dilution circuit automated an exponential growth phase followed by a 10 fold dilution, which was repeated for 500 log-growth iterations. Evolution was observed in real time as the population adapted and achieved progressively faster growth rates over time. The final evolved enzyme contained a set of 11 mutations that conferred a 90-fold improvement in substrate utilization, coinciding with the applied selective pressure. This system reduces evolution to a microfluidic algorithm, allowing the experimenter to observe and manipulate adaptation. PMID- 18399722 TI - Rethinking outreach: teaching the process of science through modeling. PMID- 18399723 TI - Progress toward public access to science. PMID- 18399725 TI - National patterns in antidepressant treatment by psychiatrists and general medical providers: results from the national comorbidity survey replication. AB - BACKGROUND: Primary care physicians, rather than psychiatrists, prescribe a majority of psychotropic medications in the United States. However, past research has shown significant differences in psychopharmacologic treatment practices of these 2 groups of physicians. The objective of this study was to compare patient characteristics and treatment patterns of adults in the United States treated with antidepressant medications by psychiatrists and other medical providers. METHOD: Data from the National Comorbidity Survey Replication (February 2001 April 2003) were used to compare characteristics of adults (aged >/= 18 years) prescribed antidepressants by psychiatrists (N = 255) or other medical providers (N = 673). The treatment groups were also compared with respect to presenting problem, antidepressant type and dose, and continuity of treatment. RESULTS: Approximately 1 in 10 adults (10.5%) were treated with an antidepressant in the past year, usually by a general medical provider (73.6%). Compared with those treated by psychiatrists, adults treated by general medical providers were significantly more likely to be at least 65 years of age and to reside in a nonurban area. By contrast, those treated by psychiatrists were significantly more likely to be male, to report significant distress, to present with serious mood or anxiety symptoms, and to meet DSM-IV criteria for mood and anxiety disorders. Individuals treated by psychiatrists typically received higher doses of medications, were less likely to stop the medication before 30 days, and were more likely to continue 90 days or longer. CONCLUSIONS: Most adults treated with anti-depressants receive the medication from general medical providers. In comparison with adults treated by psychiatrists, those treated by general medical providers are less likely to meet the criteria for mood or anxiety disorders or to continue medication beyond the first month. Quality improvement initiatives in general medical settings should focus on better targeting and continuity of antidepressant medications. PMID- 18399724 TI - Case-control analyses of the impact of pharmacotherapy on prospectively observed suicide attempts and completed suicides in bipolar disorder: findings from STEP BD. AB - OBJECTIVE: Given high rates of suicide and suicide attempts in bipolar disorder and the data suggesting a suicide-protective effect of lithium, we evaluated the impact of pharmacotherapy on prospectively observed suicides and suicide attempts in subjects in the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD). METHOD: The STEP-BD study enrolled 4360 participants with DSM-IV bipolar disorder diagnoses from September 1998 through November 2004. There were 270 suicide events in STEP-BD (8 completed suicides, 262 attempts). These occurred in 182 of STEP-BD participants (cases). Inclusion criteria required cases to be white or Caucasian, have at least 1 postbaseline visit, and have prescription information within 30 days of the suicide event. This reduced the available cohort to 106 cases. Matching included age, gender, history of previous suicide attempt, and a propensity score that considered bipolar subtype, marital status, age at onset, and history of psychosis, resulting in 93 matched pairs. A secondary analysis added mood state status within 30 days of the suicide event to the propensity score (N = 54 pairs). The association of drug prescriptions with suicide attempts/completions was assessed using a conditional logistic regression model. RESULTS: The results do not indicate a relationship between lithium use and suicide attempts or completions (p = .41). Similar findings were found for exposure to valproate, carbamazepine, lamotrigine, and the atypical antipsychotic medications. An association between selective serotonin reuptake inhibitor (SSRI) prescription and suicide events was observed (p < .0001). Findings were similar in a secondary analysis that controlled for mood state. CONCLUSIONS: Our data are not consistent with a suicide-protective effect of lithium. The association between suicide events and SSRI prescriptions requires cautious interpretation due to complex relationships between treatment, severity, and suicidality. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00012558. PMID- 18399726 TI - The impact of homocysteine levels on cognition in euthymic bipolar patients: a cross-sectional study. AB - OBJECTIVE: Bipolar disorder is associated with cognitive impairment. High homocysteine levels seem to have a negative impact on cognition in the elderly. The aim of the present study was to investigate the potential relationship of elevated homocysteine levels and cognitive impairment in bipolar patients. METHOD: Cognitive functioning of DSM-IV bipolar disorder patients who were euthymic (Hamilton Rating Scale for Depression score < or = 5 and Young Mania Rating Scale score < or = 5) and healthy controls was assessed with the revised Wechsler Adult Intelligence Scale Information Subtest, the Wechsler Adult Intelligence Scale III Letter-Number Sequencing Subtest, the Trail Making Test, and the Repeatable Battery for the Assessment of Neuropsychological Status Form A to examine premorbid IQ, information processing speed, working memory, verbal learning, visuospatial/constructional abilities, delayed memory, and executive functions. Total homocysteine plasma concentration was measured by using high performance liquid chromatography. Multivariate analyses of variance and multiple regression analyses were conducted to examine group differences and possible associations between cognitive functioning and homocysteine level. The study was conducted from 2002 through 2006. RESULTS: Seventy-five euthymic bipolar patients and 42 healthy controls participated in the study. Patients performed significantly worse than controls in all cognitive domains tested (Pillai Spur: F = 3.32, p = .038) except premorbid IQ (p = .068). The mean +/- SD homocysteine levels were 10.2 +/- 3.2 microM/L for patients and 8.9 +/- 2.8 microM/L for controls (p = .036). Stepwise regression analyses revealed a significant and independent association of homocysteine levels with verbal learning (p = .002), delayed memory (p = .030), and executive function (p = .011) in the patient group. About 11% of the variance was explained by only the homocysteine level. CONCLUSIONS: Elevated homocysteine levels may have a negative impact on verbal learning, delayed memory, and executive function in euthymic bipolar patients, but further studies are warranted. PMID- 18399727 TI - One-year course and predictors of outcome of adolescent depression: a case control study in Finland. AB - BACKGROUND: Clinical studies on the outcome of adolescent depression beyond treatment trials are scarce. OBJECTIVE: To investigate the impact of characteristics of the depressive episode and current comorbidity on the 1-year outcome of depression. METHOD: A sample of 174 consecutive adolescent psychiatric outpatients (aged 13 through 19 years) and 17 school-derived matched controls, all with unipolar depressive disorders at baseline, were reinterviewed for DSM-IV Axis I and Axis II disorders at 12 months. The study was conducted between January 1998 and May 2002. RESULTS: The outpatients had equal recovery rate and episode duration but shorter time to recurrence than the controls. Among the outpatients, Axis II comorbidity predicted shorter time to recurrence (p = .02). Longer time to recovery was predicted by earlier lifetime age at onset for depression (p = .02), poor psychosocial functioning (p = .003), depressive disorder diagnosis (p 0.5 Hz and gel concentration of 40 mg/mL fibrinogen appears to maintain cellular viability within scaffolds. Similarly, variations in gel component concentration and stimulus frequency can be modified such that a significant chondrogenic response can be achieved by hMSC in fibrin constructs after 8 h of compression spread out over 2 days. This study demonstrates the suitability of fibrin gel for supporting the cyclic compression-induced chondrogenesis of mesenchymal stem cells. PMID- 18399764 TI - The many reasons why goiter is seen in old paintings. PMID- 18399765 TI - Exogenous subclinical hyperthyroidism impairs endothelial function in nodular goiter patients. AB - BACKGROUND: Exogenous subclinical hyperthyroidism is associated with cardiovascular and metabolic changes. The aim of this study was to evaluate the effect of levothyroxine (LT4) suppression on endothelial function and insulin sensitivity in euthyroid nodular goiter patients. METHODS: Twenty-two euthyroid patients with multinodular goiter (MNG) and 22 matched healthy controls were studied. LT4 was administered in doses ranging from 50 to 150 microg/day to reach target serum thyroid-stimulating hormone (TSH) levels <0.5 mIU/L. Patients were studied before and after 8 weeks after the target TSH level <0.5 mIU/L. The control group was studied twice, 16 weeks apart. Flow mediated vasodilatation (FMD), insulin sensitivity index (ISI), lipid peroxidation, and high-sensitivity C-reactive protein (hsCRP) were the outcome measures. RESULTS: LT4 treatment significantly suppressed TSH levels to 0.2 +/- 0.1 mIU/L (minimum and maximum range was 0.05-0.3 mIU/L). FMD decreased from 10.7 +/- 2.7% to 5.4 +/- 1.7% (p < 0.001) and mean ISI decreased from 2.56 +/- 1.10 to 1.41 +/- 0.50 (p < 0.001) with LT4 treatment in the MNG group. Lipid peroxidation measured as thiobarbituric acid reactive substances (Tbars) (p < 0.05), and hsCRP (p < 0.001) levels significantly increased compared to the baseline in the MNG group. FMD measurement inversely correlated with free T4 (p = 0.008) and Tbars (p = 0.004), and positively correlated with ISI (p = 0.004). Serum Tbars and hsCRP were independent predictors of FMD (p = 0.004) in multivariate analysis. All results expressed as mean +/- SD. CONCLUSIONS: TSH suppression therapy with LT4 leading to subclinical hyperthyroidism may cause impaired endothelial function, increased oxidative stress, and decreased insulin sensitivity in euthyroid nodular goiter patients. PMID- 18399766 TI - Hypothalamic-pituitary axis and peripheral tissue responses to TRH stimulation and liothyronine suppression tests in normal subjects evaluated by current methods. AB - OBJECTIVE: To reevaluate the responses of thyrotropin-releasing hormone (TRH) stimulation test in baseline condition as well as after the administration of graded supraphysiological doses of liothyronine (L-T(3)) in normal subjects. DESIGN: To assess various parameters related to the hypothalamic-pituitary axis and peripheral tissue responses to L-T(3) in 22 normal individuals (median age: 30.5 years). Subjects were submitted to an intravenous TRH test at baseline condition and also to the oral administration of sequential and graded doses of L T(3) (50, 100, and 200 microg/day), each given over 3 days, at an outpatient clinic. Blood samples were obtained for thyrotropin (TSH) and prolactin (PRL) at basal and then 15, 30, and 60 minutes after the TRH injection. Effects of L-T(3) administration on cholesterol, creatine kinase, retinol, ferritin, and sex hormone-binding globulin (SHBG) were also measured at basal and after the oral administration of L-T(3). MAIN OUTCOME: TRH administration resulted in an increase of 4- to 14-fold rise in serum TSH (8.3 +/- 2.5-fold), and in a slight rise in serum PRL concentrations (3.8 +/- 1.5-fold). Administration of graded doses of triiodothyronine (T(3)) resulted in a dose-dependent suppression of TSH and PRL. Basal thyroxine-binding globulin (TBG) and cholesterol levels decreased, and ferritin and SHBG increased after L-T(3) administration, while creatine kinase and retinol did not change throughout the study. There was a positive correlation between basal TSH and TSH peak response to TRH at basal condition and after each sequential L-T(3) doses. On the other hand, TSH peak response to the TRH test did not predict cholesterol, TBG, ferritin, or SHBG values. CONCLUSION: Using the current methods on hormone and biochemical analysis, we standardized the response of many parameters to TRH stimulation test after sequential and graded T(3) suppression test in normal subjects. Our data suggest that the evaluation of the responses of the hypothalamus-pituitary axis to TRH test as well as the impact of L-T(3) on peripheral tissues were not modified by the current methods. PMID- 18399767 TI - Changed presentation of Hashimoto's thyroiditis in North-Eastern Sicily and Calabria (Southern Italy) based on a 31-year experience. AB - OBJECTIVE: To quantify the yearly prevalence of Hashimoto's thyroiditis at our Division and evaluate changes in its clinical presentation over the period 1975 2005. DESIGN: We have reviewed retrospectively the clinical records of patients admitted to a university hospital located in Messina, Sicily (Italy), from January 1, 1975, through December 31, 2005. This facility attracts patients from North-Eastern Sicily and most of Calabria, the Italian region across the Straits. HT was diagnosed based on various combinations of clinical, laboratory, and instrumental findings. The study group consisted of 4064 HT patients diagnosed during these 31 years. MAIN OUTCOME: We evaluated these indexes on a yearly basis: number of new HT diagnoses; age at presentation; male-to-female ratio; functional status; goitrous or nongoitrous variants with or without nodule(s); above-normal titers or levels of serum thyroid autoantibodies (thyroglobulin antibodies and thyroperoxidase antibodies). Several indexes have changed over those 31 years. Particularly, HT has become 10 times more common than it was until the early 1990s and males are relatively more represented. Patients are relatively younger, with a nongoitrous thyroid; the rate of S-Hypo exceeds largely the rate of O-Hypo. Serum thyroid autoantibodies have become less frequently positive, and when positive, they are present at a lower concentration. CONCLUSIONS: Only environmental changes, as opposed to genetic changes, can account for such alterations in the presentation of HT in our geographical area. PMID- 18399768 TI - Treating iodine deficiency: long-term effects of iodine repletion on growth and pubertal development in school-age children. AB - BACKGROUND: Iodine deficiency (ID) is still a major universal health problem. Iodine deficiency disorders (IDDs) affect people of all ages, among whom the most vulnerable are children and adolescents. The aim of the present study was to assess the long-term effects on growth and pubertal development of correcting severe ID in areas of Azerbaijan between 1999 and 2000. METHODS: Iodized oil was administered orally to 293,000 children, aged 6-16 years. Among those, 364 children were randomly selected and were examined 1 year before the administration of iodized oil (Group I-neg, iodine negative) and 295 children (Group I-Rx, iodine treated) were examined 4 years (Group I-R x 4, iodine treated 4 years later; n = 173) or 5 years (Group I-R x 5, iodine treated 5 years later; n = 122) after the last dose of iodide. RESULTS: In Group I-neg the median urine iodine concentration (UIC) (mcg/L) was 36 (mean: 36.272 +/- 11.036) and increased significantly (p < 0.001) in Group I-R x 4: 188 (mean: 230.969 +/- 155.818) and in Group I-R x 5: 175 (mean: 201.176 +/- 130.369). The prevalence of goiter was 99% in Group I-neg and 2% in Group I-R x 4. Children in Group I-Rx had a greater standard deviation score (SDS) for height (-0.1364 +/- 1.279, n = 294) than children in Group I-neg (-0.5019 +/- 1.17, n = 363) (p < 0.001, t = -3.817), which was more significant for boys. SDS for weight was similar in both groups (Group I-neg: -0.17 +/- 0.78, n = 363; Group I-Rx: -0.115 +/- 0.917, n = 294). The rate of puberty development as judged by the development of breast and pubic hair was normalized in both sexes after the correction of ID. CONCLUSIONS: Our results demonstrate that long-term correction of severe ID leads to sustained improvement of linear growth accompanied by a normalization of the time of onset of pubertal development for both sexes. PMID- 18399769 TI - The effects of recombinant TSH on bone turnover markers and serum osteoprotegerin and RANKL levels. AB - OBJECTIVE: Recently it was found that thyrotropin (TSH) receptors are present both in osteoclast and osteoblast and that TSH can modulate bone remodeling independent of thyroid hormones. The aim of this study was, firstly, to evaluate the effects of acute administration of TSH on bone remodeling markers both in men and in women and, secondly, to evaluate if these effects are mediated by variations in serum osteoprotegerin (OPG) and receptor activator of nuclear factor-KB ligand (RANKL). DESIGN: We studied 30 thyroidectomized patients (10 premenopausal and 10 postmenopausal women, 10 men) affected by thyroid carcinoma on l-thyroxine therapy. Eighty age- and sex-matched subjects were used as controls. A blood sample was drawn from each patient at baseline and 3 and 5 days after recombinant human TSH (rhTSH) administration, in preparation for (131)I whole body scan, to assess serum bone markers and serum OPG and RANKL levels. MAIN OUTCOME: At baseline, postmenopausal women and men had significantly higher values of bone turnover markers and serum OPG compared to control subjects. In all thyroidectomized patients serum RANKL was lower than in controls. After rhTSH administration, serum N-terminal propeptide of type-I procollagen (PINP), a marker of bone formation, increased significantly in postmenopausal women, while serum RANKL significantly increased after 3 days in postmenopausal patients and men returning to baseline values at day 5. Serum OPG levels did not change significantly. CONCLUSIONS: The low serum TSH observed in thyroidectomized patients on l-thyroxine therapy is associated with an increase of bone turnover in postmenopausal women and men that is associated with an increase of OPG and a decrease of serum RANKL levels. The acute TSH administration results in an increase of PINP, an index of osteoblastic activity, associated with an increase of serum RANKL. The lack of this response in premenopausal women suggests an influence of estrogen status on bone reactivity to TSH. PMID- 18399770 TI - Plasma exchanges overcome persistent iodine overload to enable 131I ablation of differentiated thyroid carcinoma. AB - BACKGROUND: Amiodarone has a high iodine content that can induce persistent iodine excess and may prevent radioiodine (RI) treatment. PATIENT: A 55-year-old obese man had taken amiodarone (200 mg/d) for 3 years and stopped 2 years earlier. He underwent total thyroidectomy for papillary cancer with extrathyroidal extension and a metastatic central lymph node, requiring RI treatment. But iodine overload, with no other documented iodinated drug intake, was found (urinary iodine excretion = 472 microg/24 h; normal < 150 microg/24 h), and persisted 3 months later. Plasma exchanges (PE) were prescribed. INTERVENTIONS AND RESULTS: Eight PE over 4 weeks were needed to eliminate 39,295 nmol of iodine. Urinary iodine excretion and serum iodine concentrations, before PE and after eight sessions were, respectively: 230 and 84 nmol/mmol of creatinine, and 811 and 71 nmol/L, enabling RI treatment (4 GBq (131)I). Post therapy whole-body scan revealed cervical uptake (0.48% of the total administered dose) corresponding to usual thyroid remnants. Ablation efficacy was confirmed 6 and 24 months later by cervical ultrasonography combined with an undetectable serum thyroglobulin level after recombinant human thyrotropin stimulation. CONCLUSIONS: When spontaneous iodine elimination is too slow to allow RI treatment of high-risk thyroid carcinoma within a reasonable time after thyroidectomy, PE are reliable and effective to overcome iodine overload. PMID- 18399771 TI - Giant posterior mediastinal parathyroid adenoma. PMID- 18399772 TI - Soft tissue implantation of thyroid adenomatous hyperplasia after endoscopic thyroid surgery. PMID- 18399774 TI - Nutrition and mutagenesis. AB - Diet-related mutagenesis plays an etiologic role in chronic diseases, including cardiovascular disease and cancer. Many dietary mutagens are DNA reactive, leading to distinct spectra of base-pair substitution mutations and structural chromosome changes. Examples include aflatoxin B1, ochratoxin A, ptaquiloside, various pyrrolizidine alkaloids, heterocyclic amines including 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine, and polycyclic aromatic hydrocarbons such as benzo[a]pyrene. However, endogenously or exogenously formed reactive species, inhibitors of topoisomerase II enzymes (e.g., flavonoids), of DNA repair (e.g., caffeine), or of the mitotic spindle (possibly acrylamide), also cause mutations, including structural chromosome changes and copy number variants. Genomic instability also results from inadequate nutrient intake (e.g., folate and selenium). Antimutagens include vitamin C, carotenoids, chlorophyllin, dietary fibers, and plant polyphenols acting through various mechanisms. Polymorphisms in genes for nutrient uptake, metabolism, and excretion will affect dietary intake in determining individual risk of disease development. Human studies utilizing nutrigenomic/nutrigenetic technologies will be essential to quantifying and overcoming diet-related mutagenesis. PMID- 18399775 TI - Ehrlichia chaffeensis infection in dogs in South Korea. AB - Ehrlichia chaffeensis is one of the causative agents of canine ehrlichiosis and human monocytic ehrlichiosis (HME). Canine ehrlichiosis caused by E. chaffeensis was diagnosed in two dogs in South Korea based on clinical findings, and the diagnosis was confirmed by polymerase chain reaction (PCR) and DNA sequencing. A 5-year-old intact male American Pit bull terrier allowed outdoors was found to be concurrently infected with Babesia gibsoni and E. chaffeensis. The major clinical findings were lethargy and reddish urine, and laboratory analysis revealed severe hematuria and thrombocytopenia. In addition, a 3-year-old neutered male Shih-tzu was also found to be infected with E. chaffeensis. Although this dog was an indoor companion animal, he was frequently allowed outside for exercise. The clinical signs observed in this dog included generalized purpura with petechiae and ecchymoses due to thrombocytopenia. A 390-bp partial portion of E. chaffeensis 16S rRNA gene was amplified in both cases, and nucleotide sequence analysis revealed 99% homology of this fragment with other E. chaffeensis isolates. These findings demonstrate the presence of E. chaffeensis infection in dogs in South Korea, and this is the first report to confirm clinical cases of E. chaffeensis infection in dogs. PMID- 18399776 TI - Presence of multiple variants of Borrelia burgdorferi in the natural reservoir Peromyscus leucopus throughout a transmission season. AB - White-footed mice (Peromyscus leucopus) serve as the principal reservoir for Borrelia burgdorferi and have been shown to remain infected for life. Complex infections with multiple genetic variants of B. burgdorferi occur in mice through multiple exposures to infected ticks or through exposure to ticks infected with multiple variants of B. burgdorferi. Using a combination of cloning and single strand conformation polymorphism (SSCP), B. burgdorferi ospC variation was assessed in serial samples collected from individual P. leucopus during a single transmission season. In individuals with ospC variation, at least seven ospC variants were recognized at each time point. One to four of these variants predominated at each time point; however, the predominant variants seldom remained consistent in an individual mouse throughout the entire sampling period. These results confirmed that mice in southern Maryland were persistently infected with multiple variants of B. burgdorferi throughout the transmission season. However, the presence of multiple ospC variants and the fluctuations in the frequency of these variants indicates that either new ospC variants are regularly introduced to this mouse population and predominate while the existing infections are cleared, or that the variation detected in the genetic profile at different time points reflects a complex mixture of B. burgdorferi populations whose relative frequencies may continually change. PMID- 18399777 TI - Prevalence of West Nile virus neutralizing antibodies in Spain is related to the behavior of migratory birds. AB - West Nile virus (WNV) is a bird flavivirus capable of infecting horses and humans that is transmitted by blood-sucking vectors. In Europe and Africa, sporadic infections and outbreaks causing human illness and deaths have occurred and have led to 2 mutually nonexclusive hypotheses regarding the circulation of WNV in Europe: (1) the occurrence of endemic sylvatic cycles that occasionally result in human or equine infection, or (2) sporadic seeding of WNV by migratory birds from areas where the virus is endemic in Africa or elsewhere that cause local epizootic foci and eventually lead to infection in humans. To investigate these 2 possibilities, we used a micro virus-neutralization test to examine the prevalence of WNV neutralizing antibodies in 574 individuals belonging to 25 species of birds captured in spring 2004 in Seville (southern Spain). Trans Saharan migrant species had both higher prevalences and antibody titers than resident and short-distance migrants. This result suggests that trans-Saharan migrants spend part of their life cycles in areas with greater circulation of WNV, or a closely related flavivirus, before their arrival in Spain. On the other hand, seroprevalences assessed in resident birds suggest a low level of WNV circulation in the studied locality. Aside from the question of local circulation, it thus seems that the risk for introduction of strains of WNV from Africa by migratory birds merits further field and experimental studies in Spain. PMID- 18399780 TI - Detection of Trypanosoma brucei in field-captured tsetse flies and identification of host species fed on by the infected flies. AB - The prevalence of trypanosome infections in tsetse flies in the Chiawa area of Lower Zambezi in Zambia, with endemic trypanosomosis, was determined by a polymerase chain reaction (PCR) method that allowed the detection of trypanosome DNA and determination of the type of animal host fed on by the tsetse fly Glossina pallidipes, using tsetse-derived DNA extracts as templates. Ninety G. pallidipes (82 females and 8 males; 18.3%) of the 492 flies captured by baited biconical traps tested positive for the presence of Trypanosoma brucei species genomic DNA. Of the 90 T. brucei-positive flies, 47 (52.2%) also tested positive for vertebrate mitochondrial DNA. Sequence analysis of the vertebrate mitochondrial DNA amplicons established that they originated from 8 different vertebrate species, namely, human (Homo sapiens), African elephant (Loxodonta cyclotis), African buffalo (Syncerus caffer), waterbuck (Kobus ellipsiprymnus), roan antelope (Hippotragus equinus), greater kudu (Tragelaphus strepsiceros), warthog (Phacochoerus africanus), and goat (Capra hircus). Furthermore, to investigate the prevalence of trypanosome infections in domestic goats in the same area where trypanosomes had been detected in tsetse files, a total of 86 goats were randomly selected from 6 different herds. Among the selected goats, 36 (41.9%) were found to be positive for T. brucei species. This combined detection method would be an ideal approach not only for mass screening for infection prevalence in tsetse populations, but also for the prediction of natural reservoirs in areas endemic for trypanosomosis. PMID- 18399779 TI - Identification of Rickettsia felis in the salivary glands of cat fleas. AB - Rickettsia felis, a flea-associated rickettsial pathogen, has been identified in many tissues, including the digestive and reproductive tissues, within the cat flea, Ctenocephalides felis. We utilized transmission electron microscopy and polymerase chain reaction to identify R. felis in the salivary glands of fed fleas and further define the distribution of R. felis within the arthropod host. We identified Rickettsia-like organisms in salivary glands using electron microscopy. Sequence analysis of portions of the Rickettsia genus-specific 17-kDa antigen gene and R. felis plasmid confirmed the morphological identification of R. felis in cat flea salivary glands. This is the first report of R. felis in tissues critical for horizontal transmission of rickettsiae. PMID- 18399781 TI - Clinical investigation of hospitalized human cases of West Nile virus infection in Houston, Texas, 2002-2004. AB - The objective of this study was to describe the clinical features of cases hospitalized with West Nile virus (WNV) infections and identify clinical parameters that could potentially predict poor outcome (death). Retrospective medical chart reviews were completed for 172 confirmed cases of WNV infection hospitalized in the Houston, Texas, metropolitan area between 2002 and 2004. Of the 172 patients, 113 had encephalitis which resulted in 17 deaths, 47 had meningitis, and 12 had uncomplicated fever. Risk factors associated with progression from encephalitis to death were absence of pleocytosis in the cerebrospinal fluid, renal insufficiency, requiring intubation and mechanical ventilation, presence of myoclonus or tremors, and loss of consciousness. These findings can aid physicians in evaluating their patients suspected of WNV infection and determining outcomes in their patients with confirmed WNV neuroinvasive disease. PMID- 18399782 TI - Bartonella spp. infection in rodents from different habitats in the Mazury Lake District, Northeast Poland. AB - Four rodent species (Clethrionomys glareolus, Apodemus flavicollis, Microtus arvalis, M. oeconomus) were captured in the period 2004-2006 in the Mazury Lake District, Northeast Poland, to determine the prevalence and genetic diversity of Bartonella species. The presence of bartonellae was assessed using polymerase chain reaction (PCR) with primers CS140f and BhCS1137n, amplifying a fragment of the gltA gene. Bartonella DNA was detected in 313 (30.6%) of 1024 rodents sampled: in 181 C. glareolus, 68 A. flavicollis, 50 M. arvalis, and 14 M. oeconomus, representing prevalence of 31.0%, 42.2%, 32.9%, and 11.1%, respectively. Comparison of the Bartonella gltA gene sequences from 38 isolates revealed six phylogenetic subgroups, out of 15 unique gltA sequences, and therein from one to five genotypic variants with homology of 88.6-99.1%. Six of 13 (46.2%) isolates from C. glareolus were identical to B. grahamii, species associated with human illness. These results have important public health implication, notably in relation to the risk of infection in humans following exposure to rodent bartonellae. PMID- 18399783 TI - Community-acquired methicillin-resistant Staphylococcus aureus emerging as an important cause of necrotizing fasciitis. AB - BACKGROUND: Necrotizing fasciitis (NF) is an uncommon fulminant soft tissue infection characterized by extensive fascial necrosis. Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) isolates producing the Panton-Valentine leukocidin (PVL) cytotoxin have been associated with serious necrotizing infections, but NF caused by CA-MRSA has been described only recently. We reviewed our NF experience at Denver Health Medical Center, where CA MRSA accounts for more than 50% of community S. aureus clinical isolates. METHODS: Patients treated for NF from January 2004 to February 2006 were identified by review of pathology records and diagnostic codes, and their medical records were reviewed. Isolates of MRSA from monomicrobial NF underwent testing for the PVL gene and pulsed-field gel electrophoresis to determine relatedness to CA-MRSA strains. RESULTS: Five of 30 NF cases during the study period, all involving the extremities, were caused by MRSA. Monomicrobial MRSA NF accounted for three cases, with all of the patients reporting a distinct "spider bite" lesion 2-3 days prior to admission. The median age was 32 years (range 28-55 years). Resistance to erythromycin and levofloxacin was present in four isolates. None of the isolates displayed inducible clindamycin resistance. Within 12 hours of admission, all patients received empiric antibiotics to which their isolate was susceptible. Patients required a median of six surgical procedures (range 2-7 operations). All patients survived. The MRSA isolates tested positive for PVL and had the USA 300 CA-MRSA deoxyribonucleic acid banding pattern. CONCLUSIONS: Community-acquired MRSA is an important cause of NF in our region, accounting for > 15% of NF cases. This infection was associated with significant morbidity necessitating multiple surgical interventions. Given the propensity of PVL positive CA-MRSA to cause severe necrotizing infections, it is reasonable to administer empiric MRSA coverage for NF in endemic locations. PMID- 18399784 TI - Abstracts from the 3rd International Congress on Complementary Medicine Research, March 29-31, 2008, Sydney, Australia. PMID- 18399786 TI - Characterization and genetic manipulation of human umbilical cord vein mesenchymal stem cells: potential application in cell-based gene therapy. AB - Stem cells are defined by two main characteristics: self-renewal capacity and commitment to multi-lineage differentiation. The cells have a great therapeutic potential in repopulating damaged tissues as well as being genetically manipulated and used in cell-based gene therapy. Umbilical cord vein is a readily available and inexpensive source of stem cells that are capable of generating various cell types. Despite the recent isolation of human umbilical cord vein mesenchymal stem cells (UVMSC), the self-renewal capacity and the potential clinical application of the cells are not well known. In the present study, we have successfully isolated and cultured human UVMSCs. Our data further revealed that the isolated cells express the self-renewal genes Oct-4, Nanog, ZFX, Bmi-1, and Nucleostemin; but not Zic-3, Hoxb-4, TCL-1, Tbx-3 and Esrrb. In addition, our immunocytochemistry results revealed the expression of SSEA-4, but not SSEA-3, TRA-1-60, and TRA-1-81 embryonic stem cell surface markers in the cells. Also, we were able to transfect the cells with a reporter, enhanced green fluorescent protein (EGFP), and a therapeutic human brain-derived neurotrophic factor (hBDNF) gene by means of electroporation and obtained a stable cell line, which could constantly express both transgenes. The latter data provide further evidence on the usefulness of umbilical cord vein mesenchymal stem cells as a readily available source of stem cells, which could be genetically manipulated and used in cell-based gene therapy applications. PMID- 18399788 TI - Looking to the future. PMID- 18399787 TI - Cyclic mechanical preconditioning improves engineered muscle contraction. AB - The inability to engineer clinically relevant functional muscle tissue remains a major hurdle to successful skeletal muscle reconstructive procedures. This article describes an in vitro preconditioning protocol that improves the contractility of engineered skeletal muscle after implantation in vivo. Primary human muscle precursor cells (MPCs) were seeded onto collagen-based acellular tissue scaffolds and subjected to cyclic strain in a computer-controlled bioreactor system. Control constructs (static culture conditions) were run in parallel. Bioreactor preconditioning produced viable muscle tissue constructs with unidirectional orientation within 5 days, and in vitro-engineered constructs were capable of generating contractile responses after 3 weeks of bioreactor preconditioning. MPC-seeded constructs preconditioned in the bioreactor for 1 week were also implanted onto the latissimus dorsi muscle of athymic mice. Analysis of tissue constructs retrieved 1 to 4 weeks postimplantation showed that bioreactor-preconditioned constructs, but not statically cultured control tissues, generated tetanic and twitch contractile responses with a specific force of 1% and 10%, respectively, of that observed on native latissimus dorsi. To our knowledge, this is the largest force generated for tissue-engineered skeletal muscle on an acellular scaffold. This finding has important implications to the application of tissue engineering and regenerative medicine to skeletal muscle replacement and reconstruction. PMID- 18399789 TI - An interview with Uwe Strahle, Ph.D. Interviewed by Vicki Glaser. PMID- 18399790 TI - Views on four key questions about zebrafish research. PMID- 18399791 TI - Initiation of a zebrafish blastula cell line on rainbow trout stromal cells and subsequent development under feeder-free conditions into a cell line, ZEB2J. AB - A continuous cell line, ZEB2, was developed from zebrafish blastula-stage embryos expressing enhanced green fluorescent protein (GFP). Originally the rainbow trout spleen cell line, RTS34st, was used as feeders to initiate and maintain the cells through several passages. ZEB2 was then grown for 2 years without feeders in L-15 with 15% fetal bovine serum (FBS) for 120 population doublings. This new cell line, ZEB2J, was heteroploid, had detectable telomerase activity, and was adherent. After growing into monolayers, some cells continued to grow into mounds. Cultures expressed Pou-2 mRNA and contained many alkaline phosphatase and a few stage-specific embryonic antigen-1-positive cells. In dishes coated with a phospholipid polymer (2-methacryloxyloxyethyl phosphorylcholine, MPC), ZEB2J formed spherical aggregates. Aggregates attached to conventional culture plastic, and most cells that emerged from aggregates had typical epithelial-like shapes of ZEB2J, which suggests that ZEB2J had limited differentiation potential, despite expressing some stem cell properties. The fluorescence of ZEB2J allowed relationships with feeder cells to be studied. In MPC dishes, ZEB2J formed mixed spheroids with RTS34st. In adherent cocultures, RTS34st and other fish cell lines strongly stimulated the ZEB2J growth, which could be quantified specifically because ZEB2J expressed GFP. ZEB2J should be useful for optimizing culture conditions for zebrafish embryonic stem cells. PMID- 18399795 TI - Catecholaminergic polymorphic ventricular tachycardia-related mutations R33Q and L167H alter calcium sensitivity of human cardiac calsequestrin. AB - Two missense mutations, R33Q and L167H, of hCASQ2 (human cardiac calsequestrin), a protein segregated to the lumen of the sarcoplasmic reticulum, are linked to the autosomal recessive form of CPVT (catecholaminergic polymorphic ventricular tachycardia). The effects of these mutations on the conformational, stability and Ca(2+) sensitivity properties of hCASQ2, were investigated. Recombinant WT (wild type) and mutant CASQ2s were purified to homogeneity and characterized by spectroscopic (CD and fluorescence) and biochemical (size-exclusion chromatography and limited proteolysis) methods at 500 and 100 mM KCl, with or without Ca(2+) at a physiological intraluminal concentration of 1 mM; Ca(2+) induced polymerization properties were studied by turbidimetry. In the absence of Ca(2+), mutations did not alter the conformation of monomeric CASQ2. For L167H only, at 100 mM KCl, emission fluorescence changes suggested tertiary structure alterations. Limited proteolysis showed that amino acid substitutions enhanced the conformational flexibility of CASQ2 mutants, which became more susceptible to tryptic cleavage, in the order L167H>R33Q>WT. Ca(2+) at a concentration of 1 mM amplified such differences: Ca(2+) stabilized WT CASQ2 against urea denaturation and tryptic cleavage, whereas this effect was reduced in R33Q and absent in L167H. Increasing [Ca(2+)] induced polymerization and precipitation of R33Q, but not that of L167H, which was insensitive to Ca(2+). Based on CASQ2 models, we propose that the Arg(33)-->Gln exchange made the Ca(2+)-dependent formation of front-to-front dimers more difficult, whereas the Leu(167)-->His replacement almost completely inhibited back-to-back dimer interactions. Initial molecular events of CPVT pathogenesis begin to unveil and appear to be different depending upon the specific CASQ2 mutation. PMID- 18399796 TI - Accumulation of free complex-type N-glycans in MKN7 and MKN45 stomach cancer cells. AB - During the N-glycosylation reaction, it has been shown that 'free' N-glycans are generated either from lipid-linked oligosaccharides or from misfolded glycoproteins. In both cases, occurrence of high mannose-type free glycans is well-documented, and the molecular mechanism for their catabolism in the cytosol has been studied. On the other hand, little, if anything, is known with regard to the accumulation of more processed, complex-type free oligosaccharides in the cytosol of mammalian cells. During the course of comprehensive analysis of N glycans in cancer cell membrane fractions [Naka et al. (2006) J. Proteome Res. 5, 88-97], we found that a significant amount of unusual, complex-type free N glycans were accumulated in the stomach cancer-derived cell lines, MKN7 and MKN45. The most abundant and characteristic glycan found in these cells was determined to be NeuAcalpha2-6Galbeta1-4GlcNAcbeta1-2Manalpha1-3Manbeta1-4GlcNAc. Biochemical analyses indicated that those glycans found were cytosolic glycans derived from lysosomes due to low integrity of the lysosomal membrane. Since the accumulation of these free N-glycans was specific to only two cell lines among the various cancer cell lines examined, these cytosolic N-glycans may serve as a specific biomarker for diagnosis of specific tumours. A cytosolic sialidase, Neu2, was shown to be involved in the degradation of these sialoglycans, indicating that the cytosol of mammalian cells might be equipped for metabolism of complex-type glycans. PMID- 18399797 TI - Apolipoprotein C-III: understanding an emerging cardiovascular risk factor. AB - The concurrence of visceral obesity, insulin resistance and dyslipidaemia comprises the concept of the metabolic syndrome. The metabolic syndrome is an escalating problem in developed and developing societies that tracks with the obesity epidemic. Dyslipidaemia in the metabolic syndrome is potently atherogenic and, hence, is a major risk factor for CVD (cardiovascular disease) in these subjects. It is globally characterized by hypertriglyceridaemia, near normal LDL (low-density lipoprotein)-cholesterol and low plasma HDL (high-density lipoprotein)-cholesterol. ApoC-III (apolipoprotein C-III), an important regulator of lipoprotein metabolism, is strongly associated with hypertriglyceridaemia and the progression of CVD. ApoC-III impairs the lipolysis of TRLs [triacylglycerol (triglyceride)-rich lipoproteins] by inhibiting lipoprotein lipase and the hepatic uptake of TRLs by remnant receptors. In the circulation, apoC-III is associated with TRLs and HDL, and freely exchanges among these lipoprotein particle systems. However, to fully understand the complex physiology and pathophysiology requires the application of tracer methodology and mathematical modelling. In addition, experimental evidence shows that apoC-III may also have a direct role in atherosclerosis. In the metabolic syndrome, increased apoC-III concentration, resulting from hepatic overproduction of VLDL (very-LDL) apoC-III, is strongly associated with delayed catabolism of triacylglycerols and TRLs. Several therapies pertinent to the metabolic syndrome, such as PPAR (peroxisome proliferator-activated receptor) agonists and statins, can regulate apoC-III transport in the metabolic syndrome. Regulating apoC-III metabolism may be an important new therapeutic approach to managing dyslipidaemia and CVD risk in the metabolic syndrome. PMID- 18399798 TI - Molecular pathogenesis of sialic acid storage diseases: insight gained from four missense mutations and a putative polymorphism of human sialin. AB - BACKGROUND INFORMATION: Free sialic acid storage diseases are caused by mutations of a lysosomal sialic acid transporter called sialin. We showed recently that the milder clinical form, Salla disease, and a related non-Finish case, are characterized by residual transport, whereas sialin mutants found in lethal infantile cases are inactive. In the present study, we have characterized the molecular effects of a putative polymorphism (M316I) and of four pathogenic mutations associated with either infantile (G127E and R57C) or Salla-like (G409E) phenotypes, or both (G328E). The transport activity of human sialin was analysed using a novel assay that was based on a construct without the functional lysosomal sorting motif, which is expressed at the plasma membrane. RESULTS: The lysosomal localization of human sialin was not (M316I and G328E) or only partially (R57C, G127E and G409E) affected by the missense mutations. In contrast, all pathogenic mutations abolished transport, whereas the putative M316I polymorphism induced an approx. 5-fold decrease of sialic acid transport. CONCLUSIONS: The molecular effects of the R57C and G127E mutations strengthen the conclusion that the infantile phenotype is caused by loss-of-function mutations. On the other hand, the milder severity of the heterozygous G409E patient may reflect an incomplete expression of the splicing mutation present on the second allele. In the case of the G328E mutation, found in the homozygous state in a clinically heterogeneous family, the apparent severity of the transport phenotype suggests that the genetic or environmental factors underlying this clinical heterogeneity might be protective. PMID- 18399799 TI - New thinking about presbyopia. PMID- 18399801 TI - Comparing methods of determining addition in presbyopes. AB - BACKGROUND: The use of plus lenses to compensate for the reduction in the range of accommodation associated with presbyopia, brings the near point of accommodation to a comfortable distance for near visual tasks. Our aim was to compare the tentative near addition determined using the most common procedures with the final addition prescribed in presbyopic patients. METHODS: Sixty-nine healthy subjects with a mean age of 51.0 years (range 40 to 60 years) were studied. Tentative near additions were determined using seven different techniques: dynamic retinoscopy, amplitude of accommodation (AA), age-expected addition, binocular fused cross-cylinder with and without myopisation, near duochrome, and balance of negative and positive relative accommodation. The power of the addition was then refined to arrive at the final addition. RESULTS: The mean tentative near additions were higher than the final addition for every procedure except for the fused cross-cylinder without initial myopisation and age expected addition methods. These biases were small in clinical terms (less than 0.25 D) with the exception of the AA procedure (0.34 D). The intervals between the 95% limits of agreement differed substantially and were always higher than +/ 0.50 D. CONCLUSIONS: All the techniques used displayed similar behaviour and provided a tentative addition close to the final addition. Due to the wide agreement intervals observed, the likelihood of error is high and supports the idea that any tentative addition has to be adjusted according to the particular needs of each patient. Among the methods examined here, we would recommend the age-expected procedure, as this technique produced results that correlated best with the final addition. PMID- 18399802 TI - Conservative management of Pseudomonal infectious sclerokeratitis. AB - Pseudomonal sclerokeratitis is an uncommon infection with the potential for loss of vision and the eye. A 66-year-old male contact lens wearer presented with a painful red eye diagnosed as sclerokeratitis. An intensive regimen of topical, subconjunctival and oral antibiotics as well as oral steroids was used as primary treatment. An overlying, focal collection of pus was incised and drained on two occasions. After three months of treatment, the infection had resolved and the treatment ceased. The involved sclera demonstrated a focal area of scleromalacia, which has remained stable. Infectious sclerokeratitis can be managed with conservative treatment without the need for surgical debridement and scleral grafting. PMID- 18399804 TI - George Smith 1941-2008. Associate professor in the Department of Optometry and Vision Sciences, The University of Melbourne. PMID- 18399800 TI - Restoration of accommodation: surgical options for correction of presbyopia. AB - Accommodation is a dioptric change in the power of the eye to see clearly at near. Ciliary muscle contraction causes a release in zonular tension at the lens equator, which permits the elastic capsule to mould the young lens into an accommodated form. Presbyopia, the gradual age-related loss of accommodation, occurs primarily through a gradual age-related stiffening of the lens. While there are many possible options for relieving the symptoms of presbyopia, only relatively recently has consideration been given to surgical restoration of accommodation to the presbyopic eye. To understand how this might be achieved, it is necessary to understand the accommodative anatomy, the mechanism of accommodation and the causes of presbyopia. A variety of different kinds of surgical procedures has been considered for restoring accommodation to the presbyopic eye, including surgical expansion of the sclera, using femtosecond lasers to treat the lens or with so-called accommodative intraocular lenses (IOLs). Evidence suggests that scleral expansion cannot and does not restore accommodation. Laser treatments of the lens are in their early infancy. Development and testing of accommodative IOLs are proliferating. They are designed to produce a myopic refractive change in the eye in response to ciliary muscle contraction either through a movement of an optic or through a change in surface curvature. Three general design principles are being considered. These are single optic IOLs that rely on a forward shift of the optic, dual optic IOLs that rely on an increased separation between the two optics, or IOLs that permit a change in surface curvature to produce an increase in optical power in response to ciliary muscle contraction. Several of these different IOLs are available and being used clinically, while many are still in research and development. PMID- 18399805 TI - A more coherent relationship between optical coherence tomography scans and retinal anatomy. PMID- 18399807 TI - Immunohistochemical characteristics of melanoma. AB - Melanoma has a wide spectrum of histologic features which mimic epithelial, hematologic, mesenchymal, and neural tumors. Immunohistochemistry has been the primary tool to distinguish melanomas from these other tumors; it has also been studied for use as an adjunct to distinguish benign and malignant melanocytic tumors and to elucidate prognosis. Furthermore, there has been extensive effort to find a suitable marker to differentiate spindle cell and desmoplastic melanoma from other tumors. We have reviewed the literature investigating melanocytic differentiation markers, proliferation markers, immunomodulatory markers, signaling molecules, and nerve growth factors and receptors. Despite the proliferation of immunohistochemical markers, S-100 remains the most sensitive marker for melanocytic lesions, while markers such as HMB-45, MART-1/Melan-A, tyrosinase, and MITF demonstrate relatively good specificity but not as good sensitivity as S-100. No marker has proven useful in distinguishing spindle cell and desmoplastic melanomas from other tumors. Ki67 remains the most useful adjunct in distinguishing benign from malignant melanocytic tumors. None of the markers reviewed has been shown conclusively to have prognostic value for melanocytic neoplasms. PMID- 18399809 TI - Bcl-2, CD34 and CD10 expression in basaloid follicular hamartoma, vellus hair hamartoma and neurofollicular hamartoma demonstrate full follicular differentiation. AB - Generalized basaloid follicular hamartoma syndrome (GBFHS) is a rare, recently described, autosomal-dominantly inherited disorder that presents with disseminated milia, palmoplantar pitting, hypotrichosis and basaloid follicular hamartomas (BFH). BFH is a benign adnexal tumor that resembles basal cell carcinoma (BCC). In this study, we report two cases of GBFHS and stain BFH, a vellus hair hamartoma (VHH) and a neurofollicular hamartoma (NH) with CD34, bcl-2 and CD10 to characterize and compare the staining patterns of these follicular tumors. Standard immunohistochemistry labeling with CD34, bcl-2 and CD10 was performed on paraffin-embedded, formalin-fixed tissue sections of five BFH (four for CD10), one VHH and one NH. CD34 stromal staining was observed in all specimens. Bcl-2 stained the outermost cell layers of the basaloid nests in all specimens. CD10 stained the peritumoral stroma of all specimens. The BFH, NFH and the VHH showed CD10 staining of matrical cells. CD34 and CD10 stain peritumoral stroma of BFH, VHH and NH. Bcl-2 stains the outermost cell layer of these tumors. CD10 was also observed to stain matrical cells. These results show the similarities in differentiation between these benign follicular neoplasms and trichoepithelioma. PMID- 18399808 TI - Collision of desmoplastic-neurotropic melanoma and squamous cell carcinoma on the lip. AB - We report on a case of the collision of a desmoplastic-neurotropic melanoma and a squamous cell carcinoma on the lip. A 46-year-old male developed a multifocal infiltrative squamous cell carcinoma of the lower lip, which also showed sparse melanocyte atypia within the epidermis and an extensive spindle cell proliferation within the dermis, subcutaneous tissues and nerves. An immunohistochemical panel showed that the spindle cells were melanocytes, not derived from the squamous cell carcinoma. Double labeling with AE1/AE3 and S100 showed striking localized proximity of the spindle-cell melanocytic and keratinocyte components in some areas of this tumor. To our knowledge, this is the first report of the collision of a squamous cell carcinoma and desmoplastic neurotropic melanoma. PMID- 18399810 TI - Circumscribed palmar hypokeratosis: two cases and a review of the literature. AB - A 60-year-old female and a 59-year-old male each presented with a solitary circumscribed patch of depressed skin of the right thenar palm and right sole, respectively. Both lesions were present for approximately 30 years without a history of trauma or other inciting incident. Histologically, the lesions showed a well-demarcated abrupt decrease in the thickness of the stratum corneum with a central area of thinned stratum corneum and hypogranulosis. Inflammation was absent. Ultrastructurally, keratin filaments were evident in the stratum corneum, and a diminished granular cell layer was apparent. There was a decrease in maturation of the keratinocytes, however, no defects in corneodesmosomes or other ultrastructural components were identified. No evidence of human papilloma virus was detected by in situ hybridization studies. Based on these 2 cases and the 25 cases reported in the literature to date, this entity seems to represent a chronic localized non-inflammatory defect in keratinization on acral sites. PMID- 18399811 TI - Eccrine gland stroma changes in cutaneous hamartoma of infancy. PMID- 18399813 TI - Quality assurance and continuing medical education in dermatopathology - the ASDP way. AB - Professional motivation led the American Society of Dermatopathology to approve the establishment of a program for quality assurance, the first of its kind in dermatopathology. Recent announcements from the American Board of Medical Specialties indicating that its member boards have a responsibility to set standards for, and to assess, the continuing competence of their diplomates have resulted in specialty board recertification programs evolving into programs of maintenance of certification. With this in mind, we initiate a CME section, administered online, in this issue of the journal based on the needs assessment of the ASDP membership survey conducted earlier. In addition to each participant receiving a printable CME certificate upon completion of the quiz, evaluation surveys are set up to enable us to get feedback essential for meeting the accreditation council for continuing medical education requirements and perhaps even more importantly to seek possible 'weaknesses' in an endeavor to improve the program. As always, your participation and constructive criticisms help us as a society to serve your needs better. PMID- 18399814 TI - Gadolinium - is it to blame? PMID- 18399815 TI - Genotypic comparison of clinical Legionella isolates and patient-related environmental isolates in The Netherlands, 2002-2006. AB - This study investigated the hypothesis that the genotype distribution of Legionella isolates from sporadic patients with Legionnaires' disease differs from that of Legionella strains in the environment. An amplified fragment-length polymorphism (AFLP) assay was used to genotype patient-derived and environmental Legionella isolates. The three Legionella pneumophila genotypes isolated most frequently from human respiratory secretions were AFLP types 004 Lyon, 010 London and 006 Copenhagen. These genotypes were cultured significantly less frequently from environmental samples (50% vs. 4%; p <0.001). The most frequently observed L. pneumophila serogroup 1 genotype among patient-derived isolates was 004 Lyon (32%). This genotype was cultured from only one of 6458 environmental samples. The positive sample contained 1.26 x 10(6) CFU/mL and originated from a whirlpool spa that had not been disinfected and had been maintained at 36 degrees C for several months. Overall, the distribution of genotypes differed significantly among patient and environmental isolates. A possible explanation is that virulent strains may exist in potential environmental sources at undetectable concentrations. PMID- 18399816 TI - First cluster of clindamycin-resistant Clostridium difficile PCR ribotype 027 in Switzerland. PMID- 18399817 TI - A gene for a new monogenic neurovascular migraine syndrome: a next step in unravelling molecular pathways for migraine? PMID- 18399818 TI - Spreading depression enhances human neocortical excitability in vitro. AB - Cortical spreading depression (CSD) plays a role in migraine with aura. However, studies of the neuronal effects of CSD in human cortex are scarce. Therefore, in the present study, the effects of CSD on the field excitatory postsynaptic potentials (fEPSP) and the induction of long-term potentiation (LTP) were investigated in human neocortical slices obtained during epilepsy surgery. CSD significantly enhanced the amplitude of fEPSP following a transient suppressive period and increased the induction of LTP in the third layer of neocortical tissues. These results indicate that CSD facilitates synaptic excitability and efficacy in human neocortical tissues, which can be assumed to contribute to hyperexcitability of neocortical tissues in patients suffering from migraine. PMID- 18399820 TI - Cluster headache prevalence: methodological considerations. PMID- 18399821 TI - The functional recurrent abdominal pain (RAP) in children may be the precursor of adult migraine. PMID- 18399828 TI - Outcomes related to placing an artificial urinary sphincter using a single incision, transverse-scrotal technique in high-risk patients. AB - OBJECTIVE: To determine, in a retrospective review, whether the single-incision technique for placing an artificial urinary sphincter (AUS) is as effective and as safe as the traditional two-incision approach in high-risk patients, as there have been questions about the efficacy and safety of a single incision, transverse-scrotal technique for placing the AUS. PATIENTS AND METHODS: From January 2000 to May 2006, 83 patients had an AUS placed using a single-incision, transverse-scrotal approach. Emphasis was put on placing the cuff around the proximal aspect of the bulbar urethra. The charts of the patients were reviewed retrospectively and the patients completed standardized questionnaires during the follow-up. RESULTS: Of the 83 patients, 24 (29%) had a history of radiation therapy and four had cryotherapy of the prostate. The mean follow-up was 18.8 months and 14 (17%) patients required revision or explantation (for any reason), nine of whom had had previous radiation therapy. The mean (sd) number of pads used per day before and after surgery was 6.7 (4.0) and 1.1 (1.6), respectively. Overall, 83% of the patients (79% of the irradiated and 85% of the unirradiated) used one or fewer pads per day after surgery. CONCLUSIONS: Even in high-risk patients there is no greater risk associated with using the single-incision, transverse-scrotal technique than in related series. Outcomes for pad use after surgery, rate of erosion, infections and mechanical malfunctions were similar to those in other series using the traditional technique. PMID- 18399827 TI - A phase II study of mifepristone (RU-486) in castration-resistant prostate cancer, with a correlative assessment of androgen-related hormones. AB - OBJECTIVE: To evaluate mifepristone (RU-486) in patients with castration resistant prostate cancer (CRPC), with a correlative assessment of serum androgens and androgen metabolites PATIENTS AND METHODS: The androgen receptor (AR) is critical in the development and progression of prostate cancer, but available antiandrogens incompletely abrogate AR signalling. Mifepristone is a potent AR antagonist that functions by competing with androgen, preventing AR coactivator binding and by enhancing binding of AR corepressors. Patients with CRPC were treated with mifepristone 200 mg/day oral until disease progression. Testosterone, dihydrotestosterone (DHT), androstenedione, dihydroepiandrosterone sulphate and the testosterone metabolite 3 alpha-diol G, were measured at baseline and during therapy. RESULTS: Nineteen patients were enrolled between April and August 2005; they were treated for a median (range) of 85 (31-338) days. The median prostate-specific antigen (PSA) level at enrollment was 22.0 (3.0-937.2) ng/mL. No patient had a PSA response (>50% reduction in PSA). Six patients had stable disease for a median of 5.5 months. After 1 month, adrenal androgens were increased and testosterone and DHT increased by 91% and 80%, respectively, compared to baseline. CONCLUSION: Mifepristone had limited activity in patients with CRPC, and stimulated a marked increase in adrenal androgens, testosterone and DHT. We hypothesise that inhibition of glucocorticoid receptor by mifepristone resulted in an increase in adrenocorticotropic hormone and subsequent increase in adrenal androgens, and that their conversion by tumour cells to testosterone and DHT probably limited the efficacy of mifepristone. These data emphasize the continued importance of alternative androgen sources in AR signalling in CRPC. PMID- 18399829 TI - Effect of a large prostate gland on open and robotically assisted laparoscopic radical prostatectomy. AB - OBJECTIVE: To evaluate the outcomes based on gland size between robotically assisted radical prostatectomy (RALP) and open RP (RRP), as larger prostates might increase the difficulty of RP. PATIENTS AND METHODS: We reviewed 660 patients who had RALP and 340 who had RRP from May 2003 to August 2006; the patients were divided into two groups, with a prostate of >75 and or=75 g had fewer positive margins than smaller glands, regardless of surgical technique. There was also a significant decrease in positive margin rate in among prostates of >75 g in favour of RALP. Thus, RALP appears to be comparable with RRP for patients with large glands, and might reduce the positive margin rate. PMID- 18399830 TI - Is robotically assisted laparoscopic radical prostatectomy less invasive than retropubic radical prostatectomy? Results from a prospective, unrandomized, comparative study. AB - OBJECTIVES: To evaluate whether robotically assisted laparoscopic prostatectomy (RALP) is less invasive than radical retropubic prostatectomy (RRP), as experimental studies suggest that the acute phase reaction is proportional to surgery-induced tissue damage. PATIENTS AND METHODS: Between May and November 2006, all patients undergoing RRP or RALP in our department were prospectively assessed. Blood samples were collected 24 h before (T0), during surgery (T1), at the end of anaesthesia (T2), and 12 (T3) and 24 h after surgery (T4), and assayed for interleukin(IL)-6 and IL-1 alpha, C-reactive protein (CRP), and lactate. The Mann-Whitney U-, Student's t- and Friedman tests were used to compare continuous variables, and the Pearson chi-square and Fisher test for categorical variables, with a two-sided P < 0.05 considered to indicate significance. RESULTS: In all, 35 and 26 patients were assessed for RALP and RRP, respectively; the median (interquartile range) age was 62 (56-68) and 68.5 (59.2-71.2) years, respectively (P < 0.009). Baseline levels (T0) of IL-1, IL-6, CRP and lactate were comparable in both arms. IL-6, CRP and lactates levels increased during both kinds of surgery. The mean IL-6 and CPR values were higher for RRP at T1 (P = 0.01 and 0.001), T2 (P = 0.001 and <0.001), T3 (P = 0.002 and <0.001) and T4 (P < 0.001 and 0.02), respectively. Lactate was higher for RRP at T2 (P = 0.001), T3 (P = 0.001) and T4 (P = 0.004), although remaining within the normal ranges. IL-1 alpha did not change at the different sample times. CONCLUSIONS: This study showed for the first time that RALP induces lower tissue trauma than RRP. PMID- 18399831 TI - A novel murine model of allogeneic vaccination against renal cancer. AB - OBJECTIVES: To develop a murine model for whole-cell allogeneic vaccination in renal cancer, as such vaccines aim to direct immune responses against patient tumour cells, due to shared antigens between the vaccine and tumour cells. MATERIALS AND METHODS: A novel murine renal cell line, allogeneic to BALB/c, was developed from a C57BL/6 mouse by primary cell culture (RVIK). It was immortalized by HPV16 E6/E7 and transfected with ras in an attempt to improve its immunogenicity. The cell line was characterized and tested as a vaccine in a BALB/c tumour-protection model after subsequent tumour challenge with autologous RenCa tumour cells. RESULTS: RVIK alone, with no ras induced cross-reactive immunity, providing a valid non-tumorigenic allogeneic whole-cell vaccine model for renal cancer. Ras transfection per se did not improve RVIK immunity. CONCLUSIONS: RVIK is a novel immunogenic murine renal epithelial cell line, which confers protection when used as an allogeneic vaccine. It provides proof of principle for the effectiveness of allogeneic whole-cell vaccines and may therefore form the basis of a useful model of allogeneic vaccination to further optimize vaccination schedules, formulation and adjuvants for a clinical setting. PMID- 18399833 TI - The drug-device continuum in the management of urological disease. PMID- 18399835 TI - Re: Effect of varicocelectomy on patients with unobstructive azoospermia and severe oligospermia. PMID- 18399836 TI - Vessel-sparing excision and primary anastomosis (for proximal bulbar urethral strictures). PMID- 18399837 TI - Quality measurement can improve the quality of care - when supervised by health professionals. PMID- 18399840 TI - Kangaroo Mother Care--a review. AB - Kangaroo Mother Care is a simple and beneficial intervention for the care of low birth weight infants. Although initially conceived for use in developing countries with limited resources, its use has expanded worldwide as clinicians, administrators and parents become familiar with the psychological, physiological, clinical and cost benefits associated with the practice. A recently documented benefit has specific relevance to blood transfusion medicine. PMID- 18399841 TI - Human platelet-specific antigens frequencies in the Argentinean population. AB - The frequencies of several human platelet antigens (HPAs) vary between different populations and are a major determinant for the prevalence of HPA alloimmunization and its clinical associated entities. The aim of this study was to characterize the allele frequencies of seven HPA systems in two different ethnic groups from the Argentinean city of Rosario, the major population and a minority Amerindian group recently arrived from the north of the country, the Tobas. A total of 192 healthy unrelated individuals from blood donors and hospital staff from the Hospital Italiano Garibaldi and 27 unrelated Toba Amerindians were genotyped for HPA-1, -2, -3, -4, -5, -6 and -15 systems by polymerase chain reaction with sequence specific primers (PCR-SSP). The present data showed that the distribution of the HPA alleles among Argentineans from Rosario is quite similar to that reported among Europeans. The frequencies seen in Tobas, although limited by the small number of aboriginal samples studied, are similar to those reported for other Amerindians populations. Statistically significant differences were found for the genotype distribution of HPA-1, -3, -5 and -15 between both groups, indicating important differences in the potential risk of HPA alloimmunization associated to transfusion and pregnancy. The study of these polymorphisms represents the first step in the elucidation of pathological conditions that are underdiagnosed in our population. It allowed us to establish a panel of characterized blood donors necessary for the serological work out and as a source for compatible platelets transfusion. PMID- 18399842 TI - Frequency of partial D in Western India. AB - Partial D is of clinical importance as the partial D-positive individuals who lack some epitopes of D antigen can develop anti-D if exposed to normal D antigen. The frequency of partial D varies in different populations. The majority of molecular studies on D variants have been reported in European, African and some East Asian populations, but no study has been reported in the Indian population so far. The aim of the study was to screen Indian population for detection of partial D by serology and classify them by multiplex polymerase chain reaction (M-PCR). The study population, consisting of 10,000 RhD-positive individuals from West India, was screened for detection of partial D using the partial D kit. In addition to these, blood samples referred because of serological RhD discrepant results from blood banks of West India were also investigated. The samples identified as partial D from these two groups were further characterized by M-PCR. Fifteen partial D cases were identified by population screening and 45 were identified from referred samples. Population screening revealed that one third of partial D was DFR when tested by partial D kit. We were able to classify 63.4 and 76.6% of partial D by partial D kit and M PCR, respectively. The incidence of partial D in West India was found to be at least 0.15% when tested with partial D kit. DFR partial D was found to be predominant in the present study. PMID- 18399843 TI - Transfusion-associated iron overload as a predictive factor for poor stem cell mobilization in patients with haematological malignancies. AB - Transfusion-associated iron overload is often observed in patients with haematological malignancies. We analysed the effect of iron overload, indicated by high serum ferritin level, on the mobilization of CD34(+) peripheral blood stem cells (PBSCs). We evaluated the association between the serum ferritin level prior to PBSC collection and the number of CD34(+) cells collected through leukapheresis in 51 patients with various haematological malignancies. Patients with serum ferritin level over 1000 ng mL(-1) were defined as high-ferritin group. Comparing the good (> or =1 x 10(6) per kg CD34(+) cells) and poor (<1 x 10(6) per kg CD34(+) cells) mobilizing groups, there was no difference in disease status, previous chemotherapies and white blood cell count at the first day of apheresis. However, there was a significant difference in the median units of red blood cell transfused between the good and poor mobilizer (2 vs. 8 units; P = 0.012). Serum ferritin level was notably higher in the poor mobilizer (1670 +/- 1320 ng mL(-1)) compared with the good mobilizer (965 +/- 705 ng mL(-1), P = 0.035). The cumulative number of CD34(+) cells per kg collected during the whole procedure was significantly lower in the high-ferritin group (5.5 +/- 4.7 x 10(6) per kg vs. 13.1 +/- 9.1 x 10(6) per kg, P = 0.01). Multivariate analysis revealed that serum ferritin level remained as an independent predictive factor for poor PBSC mobilization. Our study indicated that transfusion-associated iron overload is a predictive factor for poor PBSC mobilization. Iron chelation therapy prior to apheresis may be required to collect sufficient numbers of PBSCs in the iron overload patients. PMID- 18399844 TI - Procleix Ultrio transcription-mediated amplification vs. serological blood screening in south-western Greece. AB - We present here our overall experience after 27 months of performance of the Procleix Ultrio [HIV-1, hepatitis C virus (HCV), hepatitis B virus (HBV)] transcription-mediated amplification (TMA) assay. The aim of this report is to assess the impact of nucleic acid testing (NAT) implementation in blood screening of south-western Greek blood donors. We processed 38,264 units of blood as neat samples with the Procleix Ultrio TMA assay (Chiron/GenProbe, Emeryville/San Diego, CA, USA) between 1 January 2005 and 31 March 2007. NAT results were compared to those obtained from routine serology tests and quantitative polymerase chain reaction (PCR) assays. Overall, 52 units of blood tested positive for HBV (1.4 per thousand), 8 for HCV (0.2 per thousand) and none for HIV or multiple infections. The yield of TMA was 0.183 per thousand for HBV (7/38 264) and 0 for HCV. The TMA HBV-positive donations were tested for HBV DNA by a quantitative PCR assay and were found negative (below the detection limit of the method, 200 copies/mL). Follow-up testing showed that the TMA HBV-positive donations were positive for anti-hepatitis B core antigen immunoglobulin G antibodies. Implementation of the TMA assay in the individual donation configuration increased HBV detection compared to serological screening or a commonly used quantitative PCR assay. Follow-up studies will determine the impact of NAT implementation in HBV transmission in countries with an intermediate HBV incidence. PMID- 18399845 TI - The evidence for the use of recombinant factor VIIa in massive bleeding: development of a transfusion policy framework. AB - A review of the recent randomized control trial evidence of the use of recombinant factor VIIa (rFVIIa) in massive bleeding. rFVIIa is a recombinant genetically engineered clotting factor that has been used for the management of haemophilia patients with inhibitors. There has been increasing use in patients with massive bleeding, even when there is no underlying coagulation disorder present. In November 2006, the Canadian National Advisory Committee on Blood and Blood Products engaged in a consultation and review process with several leading Canadian experts to review and discuss the current evidence up to November 2006. There is little evidence to support the routine use of rFVIIa in massive bleeding on review of 13 randomized controlled trials. rFVIIa should only be considered as part of a transfusion policy framework for massive bleeding after all other transfusion and supportive measures are considered. An example of a policy framework is presented. PMID- 18399846 TI - How often should a red blood cell administration set be changed while a patient is being transfused? A commentary and review of the literature. AB - Current recommendations vary with regard to the frequency of change of a red blood cell (RBC) administration set. A full review was undertaken to evaluate the recommendations for how often a RBC administration set should be changed while a patient is being transfused. Comprehensive searches of Medline, Embase, Cinahl, the Cochrane Library, handsearching of transfusion journals, guidelines and websites and contact with administration set manufacturers identified 32 relevant papers: 11 clinical updates; 11 guidelines; 5 manufacturer data sheets; 3 standards; 1 Department of Health report and 1 expert opinion. Recommendations varied widely across papers. There was no pattern in recommendation by paper type, date or country of origin. Recommendations were based on change of RBC administration set either after a given number of hours or number of RBC units. The recommendations varied widely and ranged from 4 to 48 h and from 'every unit' to 'several units'. The most frequent recommendations were change of RBC administration set after 12 h or 4 units. Methodological quality of the included papers is poor. There is no formal evidence base on which to support current recommendations or challenge the current British Committee for Standards in Haematology guideline. Targeted research aimed at establishing an evidence base may be warranted and would need to document other variables that can impact frequency of change, including type of filter, age of blood and duration of RBC transfusion. PMID- 18399847 TI - Fresh frozen plasma transfusion for reversal of prolonged post-anaesthesia apnoea. AB - An 18-year old woman admitted for tonsillectomy developed prolonged post operative paralysis after anaesthesia with mivacurium. Investigation revealed a decreased cholinesterase activity because of a homozygous atypical and heterozygous K variant of the cholinesterase gene. Transfusion of fresh frozen plasma was associated with reversal of the respiratory paralysis and complete recovery. PMID- 18399848 TI - Use of TAGVHD warnings in patients receiving purine analogue chemotherapy. PMID- 18399849 TI - Haemolytic disease of the newborn presenting in a term baby born to a mother with negative red cell serology screening. PMID- 18399850 TI - Effective transfusion audit can improve and alter clinical practice: something that is often questioned. PMID- 18399851 TI - Respirology. PMID- 18399852 TI - Enhanced immediate inflammatory response to Streptococcus pneumoniae in the lungs of mice with pulmonary emphysema. AB - BACKGROUND AND OBJECTIVE: Pulmonary emphysema is associated with frequent respiratory infections but little is known about the reasons for this susceptibility to bacterial infection. We previously demonstrated an impaired inflammatory response to Streptococcus pneumoniae in an experimental emphysema mouse model at 24 h, or longer following bacterial inoculation. Toll-like receptors (TLR) have been recognized as regulators in the inflammatory response. We examined the expression of TLR on alveolar macrophages in experimental emphysema mice and evaluated the immediate inflammatory response of the emphysematous lung to streptococcal infection. METHODS: Elastase was administered once into mice trachea to induce pulmonary emphysema. Three weeks later, expression of TLR-2 and TLR-4 in the BAL cells was examined by immunostaining. Following the intratracheal inoculation of Streptococcus pneumoniae, pro inflammatory cytokine concentrations were measured in the BAL fluids of the control and emphysema mice. RESULTS: The expression of TLR-2 and TLR-4 was significantly elevated in the alveolar macrophages of emphysema mice. Six hours after infection, neutrophils in the BAL fluid of emphysema mice were significantly increased, and the levels of tumour necrosis factor-alpha, IL-1beta and IL-6 were significantly elevated, compared with the control mice. At 3 h post inoculation, macrophage inflammatory protein-2 levels were significantly elevated. CONCLUSIONS: The immediate inflammatory response in the emphysematous lung is significantly enhanced in response to streptococcal infection. This may be partly attributed to the increased expression of TLR in the alveolar macrophages of emphysema mice. PMID- 18399853 TI - Oxidant stress mediates inflammation and apoptosis in ventilator-induced lung injury. AB - BACKGROUND AND OBJECTIVE: Ventilator-induced lung injury (VILI) leads to airway epithelial cell apoptosis and lung inflammation. High tidal volume ventilation in vivo has been shown to induce MIP-2 production, lung neutrophil sequestration and apoptotic airway cell death. This study aimed to determine the effect of N acetylcysteine (NAC), a scavenger of oxygen radicals, on lung inflammation and apoptosis in an in vivo model of VILI. METHODS: Sprague-Dawley rats (n = 5 per group) were ventilated at low tidal volume (V(T) 7 mL/kg) or high tidal volume (V(T) 20 mL/kg) with or without administration of 140 mg/kg of intravenous NAC. Animals were ventilated for 30 min, 1 or 2 h, then allowed to recover for 2 h, at which time neutrophil infiltration, MIP-2, TNF-alpha and IL-6 in BAL fluid, as well as the percentage of apoptotic airway epithelial cells, were measured. RESULTS: Ventilation at V(T) 20 mL/kg increased oxidant release, as measured by serum isoprostane, and decreased lung glutathione, the major antioxidant in the lung. NAC treatment during ventilation at V(T) 20 mL/kg prevented the decrease in lung glutathione and significantly lowered serum isoprostane levels, neutrophil infiltration, cytokines in the BAL and apoptosis in the airways as compared with animals ventilated at V(T) 20 mL/kg without NAC (P < 0.05). CONCLUSIONS: These data point to an early role of oxidant-induced inflammation and apoptosis in VILI. PMID- 18399854 TI - Pleural fluid viscosity may help identifying malignant pleural effusions. AB - BACKGROUND AND OBJECTIVE: Cancer cells are larger in size and more rigid than blood cells. As the size and rigidity of cells contribute to blood viscosity, an association may exist between high pleural fluid viscosity and cancer cells in pleural effusions. The aim of this study was to determine the correlation between pleural fluid viscosity and cell constituents or laboratory data in pleural diseases with different aetiologies. METHODS: Fluid viscosities were determined in pleural effusions obtained via thoracocentesis. Pleural fluid viscosities were correlated with the laboratory data and with the percentages of different cellular constituents as assessed by cytological examination. RESULTS: Pleural fluid viscosity was highest in malignant pleural effusions with positive results on cytological examination, and was correlated with the percentages of tumour cells (Spearman's rho = 0.24, P = 0.037) and mitotic figures (rho = 0.23, P = 0.041) in the exudates. Multivariate logistic regression analysis showed that pleural fluid viscosity was a significant determinant of positive results on cytological examination (odds ratio (OR) 6.26, 95% confidence interval (CI) 1.32 29.8), as were the levels of protein (OR 1.48, 95% CI 1.01-2.16) and LDH (OR 1.001, 95% CI 1-1.002). CONCLUSION: High pleural fluid viscosity may suggest a potential diagnosis of malignant pleural effusion. PMID- 18399855 TI - Association of beta2-adrenoreceptor genotypes with bronchodilatory effect of tiotropium in COPD. AB - BACKGROUND AND OBJECTIVE: Recently, there has been interest in interactions of beta(2) adrenergic receptors (beta(2)-AR) and muscarinic acetylcholine receptors (mAChR), which share intracellular signal transduction systems. The aim of the present study was to investigate whether bronchodilator response to tiotropium is influenced by beta(2)-AR genotype in patients with COPD who show poor responsiveness to inhaled beta(2)-agonists. METHODS: After a 4-week run-in period, patients with COPD were treated with inhaled tiotropium bromide (18 microg once daily) for 8 weeks. Spirometric measurements and reversibility testing with inhaled beta(2)-AR agonists were performed and health-related quality of life was assessed using the St George's respiratory questionnaire (SGRQ) before and after treatment. Genomic DNA was prepared from peripheral blood and individual genotypes at amino acid 16 of the beta(2)-AR were examined. RESULTS: Forty-four patients with COPD completed the study. COPD patients with the Arg/Arg genotype (n = 22) had a significant increase in FEV(1) during treatment compared with those without the Arg/Arg genotype (n = 22) (FEV(1), P = 0.009; FEV(1)%, P = 0.006). While all component and total scores on the SGRQ improved significantly in both genetic groups, changes in impact and total scores were significantly greater in patients with Arg/Arg compared with those without (total scores, P = 0.005; impact scores, P < 0.001). CONCLUSIONS: These findings indicate that the homozygous Arg/Arg genotype at amino acid 16 of the beta(2)-AR could affect bronchodilator response to tiotropium in patients with COPD with significant effects on health-related quality of life. PMID- 18399856 TI - Clinical benefit of sequential three-step empirical therapy in the management of chronic cough. AB - BACKGROUND AND OBJECTIVE: A prospective study was conducted to investigate the efficacy of empirical therapy in the management of chronic cough. METHODS: A three-step empirical therapy protocol was used to treat patients who had a chronic cough and a normal CXR and normal lung function testing. In step one antihistamine decongestants plus bronchodilators were taken, corticosteroids in the second step, and a combination of a proton pump inhibitor and a prokinetic agent in the third step. Cough symptom scores were recorded before and after each step in the protocol. RESULTS: There were 102 patients recruited; mean age 49 +/- 16 years and with a history of cough of between 2 and 120 months at presentation. The cough resolved in 67.6% (n = 69) of the patients after the first-step therapy; in a further 12.7% (13) after the second-step therapy, and in another 7.8% (8) after the third-step of the treatment protocol. In three patients cough improved after the first-step of therapy but only achieved complete resolution when this treatment was combined with the third-step therapy. Overall, the three step empirical therapy was successful in 88.2% (n = 90) of the patients. Cough symptom scores decreased from 3.30 +/- 0.75 to 0.30 +/- 0.97 (P < 0.001). The mean duration of treatment was 2.70 +/- 1.52 weeks (range 1-12). Cough was controlled in <4 weeks in 79.4% (n = 81) of patients. CONCLUSIONS: Three-step empirical therapy seemed to be useful in most patients and has practical application in the management of chronic cough. PMID- 18399857 TI - Exhaled nitric oxide levels in patients with atopic cough and cough variant asthma. AB - BACKGROUND AND OBJECTIVE: Atopic cough (AC) is an established clinical entity in Japan, in which patients present with a chronic persistent non-productive cough. Exhaled nitric oxide (NO) is a biomarker of eosinophilic airway inflammation. The present study examined whether exhaled NO levels were increased in AC in comparison with cough variant asthma (CVA) and bronchial asthma (BA). METHODS: Consecutive patients presenting with an isolated cough lasting at least 8 weeks were enrolled in the study. The aetiology of the chronic cough was determined according to the Japanese Respiratory Society guidelines for management of cough. Exhaled NO, capsaicin cough sensitivity (capsaicin concentration eliciting five or more coughs (C5)) and bronchial reversibility were measured at the patients' first visit. Bronchial responsiveness (PC20 to methacholine) was measured at their second visit following a 6-day course of bronchodilator therapy. RESULTS: There were 58 patients recruited and fully investigated; of these 9 and 11 patients were diagnosed with AC and CVA, respectively, as single causes of chronic cough. Ten patients with BA who had not received corticosteroid therapy in the previous 4 weeks and who attended the same clinic in the same time period acted as controls. Exhaled NO levels in patients with AC were significantly lower than those in patients with CVA and BA. There was no significant difference in the exhaled NO levels between patients with CVA and BA. CONCLUSIONS: Exhaled NO may reflect eosinophilic inflammation of peripheral airways and its measurement may be useful in differentiating CVA from AC and other causes of chronic non productive cough. PMID- 18399858 TI - New predictive equations for spirometric reference values and comparison with Morris equation in a Korean population. AB - BACKGROUND AND OBJECTIVE: The clinical importance of the differences between actual and predicted spirometric indices in non-Western populations is poorly defined. This study evaluated the differences between the spirometric values derived from Morris equation, traditionally used in South Korea, and the actual values, in the classification and detection of patients with respiratory diseases, and developed new predictive equations for the calculation of reference spirometric values for healthy Koreans. METHODS: Data derived from a subset of the population who completed the initial baseline survey of the Korean Health and Genome Study were used to develop new predictive equations for spirometric reference values, using multiple linear regression. The effects of the new equations relative to those of Morris on the detection and classification of patients with respiratory diseases were then evaluated. RESULTS: In total, 9999 people completed the baseline survey; a subgroup of 1314 met the study inclusion criteria and were used to develop the new predictive equations. Morris equation are 53.8% less accurate in detecting people with restrictive disorders and 15.8% less accurate in estimating the severity of COPD than the newly derived equations, although the differences between values derived from the traditional equations and values from the new equations were as small as 3.3-7.6%. CONCLUSIONS: The use of spirometric reference values that underestimate the actual parameters, despite the small differences, may have a significant influence on the detection of patients with restrictive disorders and the staging of COPD. PMID- 18399859 TI - Longitudinal study of airway dimensions in chronic obstructive pulmonary disease using computed tomography. AB - BACKGROUND AND OBJECTIVE: Chest CT has been widely used for the evaluation of structural changes in lung parenchyma and airways in cross-sectional studies. There has been no report on the annual changes in airway dimensions as assessed by CT in COPD patients. The objective of this study was to investigate the annual changes in airway dimensions and lung attenuation using CT in patients with COPD and to evaluate the correlations among annual changes in CT measurements and pulmonary function. METHODS: Eighty-three men with COPD had completed five annual assessments of CT scans and pulmonary function tests over 4 years. Airway dimensions of the basal segment bronchi and lung attenuation on CT images were analysed in 38 subjects in whom the same airway could be measured at least three times, including at entry and at the end of the study. RESULTS: Mean annual decline in FEV(1) was 21 mL/year. Annual changes in the percentage of low attenuation areas were not significantly correlated with decline in FEV(1). On the other hand, annual changes in the percentage of wall area (WA%/year) were significantly inversely correlated with annual changes in FEV(1) (r = -0.363, P = 0.025), whereas WA%/year did not differ among severity stages at entry and did not correlate with baseline FEV(1). CONCLUSIONS: The results showing that annual changes in airway thickening correlated with annual decline in air flow limitation suggests the importance of treatment of airway inflammation in COPD. CT is a useful tool for quantitative estimation not only of emphysema but also of airway lesions in longitudinal studies. PMID- 18399860 TI - Effect of thixotropy conditioning of inspiratory muscles on the chest wall response to CPAP. AB - BACKGROUND AND OBJECTIVE: Thixotropy conditioning of inspiratory muscles changes the end-expiratory position of the respiratory system during resting breathing. We examined the immediate effects of thixotropy conditioning of inspiratory muscles on chest wall inflation induced by CPAP. METHODS: A cross-over design study was performed in 13 healthy men. Operating chest wall volume (Vcw) was measured by respiratory inductive plethysmography. Conditioning consisting of a 5 s inspiratory effort at a mask pressure of -20 cm H(2)O was performed under CPAP (10 cm H(2)O) at three Vcw values to change the muscle length at which conditioning occurred: (i) FRC at a CPAP of 10 cm H(2)O (FRC(10)); (ii) FRC at a CPAP of 0 cm H(2)O (FRC(0)); and (iii) RV at a CPAP of 10 cm H(2)O (RV(10)). RESULTS: CPAP (10 cm H(2)O) increased FRC by 0.60-0.70 L. Decreases in operating Vcw were noted after conditioning at RV(10), which decreased end-expiratory Vcw by 0.16 +/- 0.12 L and end-inspiratory Vcw by 0.20 +/- 0.14 L at 180 s (P < 0.01). Inspiratory capacity (1.59 +/- 0.45 L) and inspiratory reserve volume (1.18 +/- 0.40 L) were greater than preconditioning values (1.44 +/- 0.49 L and 0.99 +/- 0.41 L, respectively; P < 0.01). Conditioning at FRC(0) also decreased operating Vcw significantly, but conditioning at FRC(10) did not change operating Vcw. CONCLUSIONS: Thixotropy conditioning of inspiratory muscles performed below inflated FRC decreased the level of CPAP-induced inflation. PMID- 18399861 TI - Continuous positive airway pressure increases inspiratory capacity of COPD patients. AB - BACKGROUND AND OBJECTIVE: Hyperinflation with a decrease in inspiratory capacity (IC) is a common presentation for both unstable and stable COPD patients. As CPAP can reduce inspiratory load, possibly secondary to a reduction in hyperinflation, this study examined whether CPAP would increase IC in stable COPD patients. METHODS: Twenty-one stable COPD patients (nine emphysema, 12 chronic bronchitis) received a trial of CPAP for 5 min at 4, 7 and 11 cmH(2)O. Fast and slow VC (SVC) were measured before and after each CPAP trial. In patients in whom all three CPAP levels resulted in a decreased IC, an additional trial of CPAP at 2 cmH(2)O was conducted. For each patient, a 'best CPAP' level was defined as the one associated with the greatest IC. This pressure was then applied for an additional 10 min followed by spirometry. RESULTS: Following application of the 'best CPAP', the IC and SVC increased in 15 patients (nine emphysema, six chronic bronchitis). The mean change in IC was 159 mL (95% CI: 80-237 mL) and the mean change in SVC was 240 mL (95% CI: 97-386 mL). Among these patients, those with emphysema demonstrated a mean increase in IC of 216 mL (95% CI: 94-337 mL). Six patients (all with chronic bronchitis) did not demonstrate any improvement in IC. CONCLUSIONS: The best individualized CPAP can increase inspiratory capacity in patients with stable COPD, especially in those with emphysema. PMID- 18399862 TI - Effects of pulmonary rehabilitation in patients with idiopathic pulmonary fibrosis. AB - BACKGROUND AND OBJECTIVE: Although pulmonary rehabilitation is effective for patients with COPD, its efficacy in patients with IPF is unknown. The purpose of this study was to evaluate the effects of pulmonary rehabilitation in IPF. METHODS: Thirty patients diagnosed with IPF, according to the consensus statement, were randomly assigned to the rehabilitation group or the control group. The pulmonary rehabilitation mainly consisted of a 10-week programme of exercise training. Pulmonary function, blood gas analysis, 6MWD, dyspnoea rating with the baseline dyspnoea index and health-related quality of life score on the St George's Respiratory Questionnaire were evaluated at baseline and after the programme. RESULTS: Assessment of efficacy was carried out on 13 patients who completed the programme and 15 patients in the control group. There were no significant effects of the programme on measures of pulmonary function, values of arterial blood gas analysis or dyspnoea rating. Although there were some differences in the baseline 6MWD and total health-related quality of life score which were not statistically significant, marked improvements were observed in the 6MWD (mean difference 46.3 m (95% CI: 8.3-84.4), P < 0.05) and the total health-related quality of life score (-6.1 (95% CI: -11.7 to -0.5), P < 0.05). CONCLUSIONS: Pulmonary rehabilitation improves both exercise capacity and health related quality of life in patients with IPF. PMID- 18399863 TI - Predictors for failed dose reduction of inhaled corticosteroids in childhood asthma. AB - BACKGROUND AND OBJECTIVE: Studies of Western populations have shown that increased exhaled nitric oxide (FeNO) and/or sputum eosinophils (sp-Eos) are predictive of asthma exacerbations. However, the utility of these measurements in different populations and settings is unknown. This study aimed to determine the predictors for failure of reduction of inhaled corticosteroid (ICS) doses in children with stable asthma. METHODS: Fifty children (median age 11.8 years, interquartile range (IQR) 5.9 years) had their dose of ICS halved every 8 weeks until they reached the study end-point (exacerbation or weaned off ICS). Spirometry, FeNO and induced sputum cells were measured at baseline and at each stage of ICS reduction. RESULTS: Eleven subjects suffered an asthma exacerbation and the remainder was successfully weaned off ICS. Subjects with an exacerbation were older (15.4 years (IQR 5.4) vs 11.4 years (IQR 3.9), P = 0.019) and more likely to be boys (P = 0.035). FeNO (median 156 p.p.b. (IQR 131) vs 76.1 p.p.b. (IQR 79.5), P = 0.013) and sp-Eos (17.3% (IQR 33.8%) vs 7.1% (IQR 9.9%), P = 0.019) were significantly greater in those who had an exacerbation. The areas under the receiver operating characteristic curves for FeNO (0.78, 95% CI: 0.59 0.97, P = 0.013) and sp-Eos (0.76, 95% CI: 0.56-0.96, P = 0.016) were similar (P = 0.88) and both were significantly greater than that for FEV(1)% predicted (0.12, 95% CI: 0.08-0.56, P = 0.0013). CONCLUSIONS: Older boys with raised FeNO and sp-Eos are at higher risk of failure of reduction in their ICS dose. Monitoring airway inflammation in children with asthma using FeNO or sp-Eos is clinically useful in guiding ICS dose reduction in a non-Western outpatient setting. PMID- 18399864 TI - Clinically occult subpleural fibrosis and acute interstitial pneumonia a precursor to idiopathic pulmonary fibrosis? AB - BACKGROUND AND OBJECTIVE: The clinical features and acute component of the pathological findings in patients with acute exacerbation of IPF are similar to those of acute interstitial pneumonia (AIP). A growing body of research suggests that acute exacerbation of subclinical IPF is similar to AIP. This study tested the hypothesis that subpleural fibrosis (resembling localized usual interstitial pneumonia) in patients with diffuse alveolar damage can mimic patients with AIP alone. METHODS: The clinical, radiological and histopathological characteristics of patients with AIP with or without subpleural fibrosis, as identified at post mortem examination, were assessed. RESULTS: Fourteen patients with AIP met the inclusion criteria. These comprised nine men and five women ranging in age from 65 to 90 years. The mean duration from onset of symptoms to admission was 12.3 days, and from admission to death was 50.5 days. All patients presented with severe respiratory failure with bilateral pulmonary infiltrates on CXR. CT scans revealed bilateral ground glass opacity (100.0%), air space consolidation (69.2%) and no honeycombing. In seven patients (50.0%) histological examination showed subpleural fibrosis. There were no differences in the clinicopathological or CT scan findings between patients with AIP only compared with those with AIP and subpleural fibrosis. CONCLUSIONS: Subpleural fibrosis resembling localized usual interstitial pneumonia is often present in patients diagnosed as having AIP. These patients were not distinguishable from those with AIP only, based on the clinicopathological and radiological findings. PMID- 18399865 TI - Solid-organ malignancy as a risk factor for tuberculosis. AB - BACKGROUND AND OBJECTIVE: The effective control of tuberculosis (TB) requires that people at high risk for the reactivation of TB are identified. Haematological malignancy has been shown to be a risk factor for the development of TB, either through immune suppression by the tumour or through the effects of chemotherapy. This study assessed the hypothesis that solid-organ malignancy is a risk factor for the development of TB. METHODS: A retrospective cohort study was performed to determine the incidence of TB in patients with solid-organ malignancy and in control subjects without malignancy. Risk factors for the development of TB among patients with cancer were also assessed. RESULTS: The study recruited 1809 cases with cancer and 1809 control subjects and followed them for 3 years. The incidence of active TB per 1000 person-years was 3.07 in patients with cancer and 0.77 in controls (P = 0.009). Compared with the control group, patients with cancer had an increased risk of developing TB (incidence rate ratio (IRR) 4.69, 95% CI: 1.52-14.46). Proportional hazards regression analysis showed that the risk factors for development of TB were chronic renal failure (IRR 9.91, 95% CI: 1.17-83.60), old healed TB on CXR (IRR 45.05, 95% CI: 5.74-353.88), and anticancer chemotherapy (IRR 4.32, 95% CI: 1.10-16.89). An interaction between old healed TB and anticancer chemotherapy was observed. CONCLUSION: These findings indicate that immune suppression by cancer or by anticancer chemotherapy increases vulnerability to reactivation of TB, especially in cancer patients with old healed TB. PMID- 18399866 TI - Validation of symptom-based COPD questionnaires in Japanese subjects. AB - BACKGROUND AND OBJECTIVE: Symptom-based questionnaires may be helpful in diagnosing patients with COPD. The aim of this study was to determine whether two COPD questionnaires designed in Western countries were applicable to Japanese and other Asian patients. METHODS: The participants were Japanese people aged 40 years and over. Each subject answered questions on demographics and symptoms and underwent spirometry before and after administration of a bronchodilator. Questionnaire A was designed to identify previously undiagnosed COPD and questionnaire B was designed to differentiate between COPD and asthma. RESULTS: The numbers of COPD patients who answered questionnaires A and B were 33 of 169 (19.5%) and 112 of 168 (66.7%), respectively. Comparison of the COPD group with the non-COPD group revealed a significant difference in total score in both questionnaire A and questionnaire B (both P < 0.001). The area under the receiver operating characteristic curve (AUC-ROC) for questionnaire A was 0.791. With a cut-off value of 16.5 points, the sensitivity and specificity were 0.939 and 0.404, and with a 19.5-point cut-off, sensitivity and specificity were 0.848 and 0.647, respectively. The AUC-ROC for questionnaire B was 0.765. With cut-off values of 18.5 and 24.5 points, the respective sensitivities and specificities were 0.946 and 0.393, and 0.741 and 0.607. CONCLUSIONS: A simple self administered questionnaire can help to diagnose COPD in Japanese subjects. When these questionnaires are used in Japan, cut-off values should be set somewhat higher than in Western countries. PMID- 18399867 TI - Comparison of serological tests for detection of immunoglobulin M antibodies to Chlamydophila pneumoniae. AB - BACKGROUND AND OBJECTIVE: To evaluate an enzyme immunoassay (EIA) (AniLab C. pneumoniae) for detecting anti-Chlamydophila pneumoniae-specific IgM antibody, by comparing it with an ELISA, Hitazyme C. pneumoniae, and a micro immunofluorescence (MIF) test. METHODS: Antibodies in sera from three groups of patients were measured: eight serum samples collected serially from a patient with acute C. pneumoniae pneumonia, 34 serum samples with Hitazyme-ELISA false positive results, and 137 serum samples from patients with community-acquired pneumonia. RESULTS: The IgM antibody titre in the patient with acute C. pneumoniae pneumonia showed almost identical variation with the EIA, ELISA and MIF tests. Among the 34 samples found to be false-positive for IgM with ELISA, EIA revealed no positive cases. When a true positive case was defined as one for which a positive reaction was obtained with at least two tests, the sensitivities of the EIA, ELISA and MIF tests were 97.1%, 100% and 74.3%, with specificities of 100%, 37.3% and 100%, respectively. CONCLUSIONS: EIA was highly sensitive and specific as compared with the MIF test, and the ELISA test showed the lowest specificity. Consequently, the AniLab-EIA, rather than the Hitazyme-ELISA, is recommended as the routine method for accurately diagnosing acute C. pneumoniae infection. PMID- 18399868 TI - Diagnosis of pulmonary tuberculosis using MTB12 and 38-kDa antigens. AB - BACKGROUND AND OBJECTIVE: Mycobacterium tuberculosis MTB12 protein plays an essential role in pro-inflammatory responses during the early stages of human pulmonary tuberculosis (TB), even though the T-cell immunoreactivity of MTB12 is weaker than that of the 30-kDa antigen (Ag). The objective of this study was to evaluate the humoral immune responses induced by MTB12 Ag during human TB. METHODS: Using an ELISA, anti-MTB12 IgG levels in the sera of TB patients and healthy controls were compared with those induced by the 30-kDa Ag and 38-kDa Ag, or both. RESULTS: In TB patients, the sensitivity and specificity of MTB12 Ag were similar to those of other antigens at 53.0% and 95.4%, respectively. However, the sensitivity increased to 73.0% when the combination of MTB12 and 38 kDa Ag was measured. Specificity remained high when a combination of the individual antigens was used. ELISA results showed that after anti-tuberculosis treatment, the mean IgG levels against MTB12 alone or MTB12 plus 38-kDa Ag were significantly increased in the TB patients, while those against MTB12 plus 30-kDa Ag were not (P < 0.05). CONCLUSIONS: Collectively, these data suggest that MTB12, in combination with 38-kDa Ag, can be used to increase the accuracy of pulmonary TB diagnosis. PMID- 18399869 TI - BODE score is a useful predictor of hospital admission in rural patients with chronic obstructive pulmonary disease. AB - BACKGROUND AND OBJECTIVE: COPD is a chronic illness with frequent episodic exacerbations that require admission to hospital. The aim of this study was to identify risk factors (or predictors) for hospital admission in a sample of rural COPD patients. METHODS: COPD patients from the Goulburn and Crookwell areas of New South Wales, Australia, were included in the study. Patients were divided into two groups: those with two or fewer admissions, and those with three or more admissions in the last year. Patient interviews were conducted, and data were collected on demographics, consumption of health resources, COPD severity measures (BODE score) and quality of life. RESULTS: There were 32 patients recruited with a median age of 68 years, and 21 (65.6%) were male. There were no significant differences in risk factors (or predictors) identified between the two groups. A statistical difference was found between the two groups, with patients with three or more hospital admissions having a higher BODE score (P = 0.004), poorer quality-of-life score (P = 0.015) and reduced exercise tolerance (P = 0.001). CONCLUSIONS: In this small sample of COPD patients, there were no clear differences in terms of risk factors for COPD admission between the two groups. The BODE score was found to be a useful predictor of disease severity and likelihood of hospital admission. PMID- 18399870 TI - Influenza-associated pneumonia in a Turkish area with endemic avian influenza. AB - BACKGROUND AND OBJECTIVE: The aim of the study was to describe the effects of influenza associated with community-acquired pneumonia (CAP) on the outcome of patients in an area characterized by endemic avian influenza. METHODS: Sixty patients diagnosed with CAP and requiring hospitalization were studied for the presence of influenza. Based on the presence or absence of influenza, patients were divided into two groups: group 1 were CAP patients positive for influenza and group 2 included CAP patients negative for influenza. Laboratory investigations, CXR findings and prognosis were compared. RESULTS: Nasopharyngeal swabs or deep tracheal aspirates were tested for viral aetiology, and seven patients were positive for influenza AH3; one was positive for influenza AH1; seven were positive for influenza B; and one was positive for parainfluenza. Group 1 patients (n = 16) had a mean age of 56 years and group 2 patients (n = 45) had a mean age of 55 years. Significant increases in creatinine kinase and lymphopaenia were seen in group 1 patients. On CXR, interstitial infiltration was more marked in group 1. There was significantly higher mortality in group 1 than in group 2. CONCLUSION: CAP associated with influenza had a different clinical picture and outcome compared with patients without evidence of influenza. PMID- 18399871 TI - Stable prevalence of asthma symptoms in school-aged children in the Torres Strait region. AB - BACKGROUND AND OBJECTIVE: To (i) determine if the prevalence of asthma has altered in two previously studied communities and (ii) obtain baseline measures in two further communities in the Torres Strait region, Australia. METHODS: A population-based cross-sectional study of school-aged children was conducted. Five schools in four communities were selected: 361 children aged 5-17 years participated. The study used the same epidemiological tool that had been utilized to measure asthma prevalence (locally adapted International Study of Asthma and Allergy in Childhood questionnaire). RESULTS: The overall response rate was 30%; response rates in individual communities ranged from 23% to 100%. The prevalence of self-reported wheezing in the last 12 months decreased from 10.7% to 6.6% (P = 0.109) on Thursday Island and from 3.1% to zero (P = 0.358) on Warraber Island. The percentage of children with asthma symptoms was lower in this current study but changes were not statistically significant. Overall self-reported prevalence of ever wheezing was 12.5%; 5.4% reported wheezing in the previous 12 months, 5.9% reported wheezing after exercise and 12.2% reported ever having asthma. There was considerable inter-community variation in the prevalence of symptoms. CONCLUSIONS: Asthma prevalence in school-aged children living in the Torres Strait region remains high but, as in mainstream Australian children, the prevalence is stable. PMID- 18399872 TI - Possible therapeutic effect of direct haemoperfusion with a polymyxin B immobilized fibre column (PMX-DHP) on pulmonary oxygenation in acute exacerbations of interstitial pneumonia. AB - BACKGROUND AND OBJECTIVE: Acute exacerbations of interstitial pneumonias (IP) can occasionally occur, and have an extremely poor prognosis. Recently, direct haemoperfusion with a polymyxin B immobilized fibre column (PMX-DHP) was shown to have a beneficial effect in acute exacerbations of IPF. However, little is known about the efficacy of PMX-DHP in acute exacerbations of other IP. This study investigated the effectiveness and safety of PMX-DHP in acute exacerbations of IP. METHODS: The study subjects consisted of five patients with an acute exacerbation of IP, including three with IPF, one with idiopathic interstitial pneumonia (IIP) with atypical radiological findings of IPF, and one with myeloperoxidase antineutrophil cytoplasmic antibody (MPO-ANCA)-related IP. The patients were treated with two courses of 3-12 h each of PMX-DHP, concurrently with corticosteroids alone or plus cyclophosphamide. RESULTS: After two courses of PMX-DHP, the PaO(2)/fraction of inspired oxygen (FiO(2)) (P/F) ratio increased rapidly from an average of 93 to 260 mm Hg, and there was radiological improvement in all patients. However, one patient treated for 3 h each time eventually died of respiratory failure, and two patients treated for 6 h each time died from respiratory infections. The other two patients were treated for 12 h each time, and the therapeutic effects lasted longer, with both surviving longer than 48 days. No adverse effects were detected apart from thrombocytopaenia. CONCLUSION: PMX-DHP therapy was safe and effective in improving oxygenation in acute exacerbations of IPs, either with corticosteroids alone or plus cyclophosphamide, and may be beneficial for the treatment of this condition. PMID- 18399873 TI - Safety and efficacy of using a surgivac pump for the drainage of chronic indwelling pleural catheters in malignant pleural effusions. AB - BACKGROUND AND OBJECTIVE: This descriptive study assessed whether a reusable commercially available surgivac pump was a safe and affordable method for draining chronic malignant pleural effusions with an indwelling pleural catheter. METHODS: Patients who were managed as outpatients using this technique were recruited over a 5-year period in Cairo, Egypt. The indwelling pleural catheters were inserted under local anaesthesia in a bronchoscopy suite. Patients were instructed by a trained nurse on how to drain the catheter using the surgivac pump. RESULTS: Fifty-five patients were included in the study. Successful pleurodesis was achieved in 42 (76.3%) over a mean period of drainage of 19.1 days (range 12-59 days). In all patients, the surgivac pump was successful in draining their pleural fluid and there were no complications related to the device itself. CONCLUSION: The use of a surgivac pump to drain malignant effusions via a chronic indwelling pleural catheter is safe and results in a pleurodesis comparable to the more commonly used negative pressure containers (vacuum bottles). PMID- 18399874 TI - Peritoneal carcinomatosis in lung cancer. AB - BACKGROUND AND OBJECTIVE: A survey was carried out of peritoneal carcinomatosis (PC) in lung cancer. METHODS: A retrospective analysis was conducted of all lung cancer patients with malignant ascites diagnosed between 1990 and 2005 at a general hospital in Taiwan. RESULTS: There were 30 lung cancer patients with documented PC. The most frequent abdominal symptom was abdominal distension. The most common histological type of lung cancer with PC was adenocarcinoma. Eighty per cent of the patients developed malignant pleural effusions before the diagnosis of PC. The median survival after diagnosis of PC was 15 days. Two adenocarcinoma patients with PC benefited from treatment with gefitinib. CONCLUSIONS: Further study is warranted to investigate the use of gefitinib in treatment of patients with lung adenocarcinoma and PC. PMID- 18399875 TI - Interferon-gamma responses after isoniazid chemotherapy for latent tuberculosis. AB - BACKGROUND AND OBJECTIVE: Chemotherapy for Mycobacterium tuberculosis infection may decrease interferon (IFN)-gamma responses to early secretory antigenic target 6 and culture filtrate protein (CFP)-10; a reaction that could be useful to monitor the success of treatment. We investigated IFN-gamma responses in subjects with latent TB infection before and after isoniazid (INH) chemotherapy. METHODS: A total of 48 patients who had contact with a TB patient in a psychiatric hospital were suspected to have latent TB infection on the basis of a positive QuantiFERON-TB Gold (QFT-G) test and were offered INH treatment for 6 months. After INH chemotherapy, IFN-gamma responses were again quantified and compared with initial measurements. RESULTS: Thirty-four patients completed 6 months of therapy and 28 were retested. Seven (25%) had a negative test and the other patients showed an overall decline. Geometric mean for early secretory antigenic target 6 decreased from 1.398 to 0.362 (P < 0.001), and that for CFP-10 from 0.312 to 0.120 (P < 0.001). A subsequent QFT-G test carried out 18 months after chemotherapy showed no further decline of IFN-gamma responses. CONCLUSIONS: If the success of chemotherapy is defined as negative conversion in the QFT-G test, these results suggest that although the waning of QFT-G responses as a result of chemotherapy is an important characteristic of IFN-gamma responses, the extent of waning would not be sufficient to allow effective monitoring of the success of chemotherapy because a majority of contacts still showed positive responses in the QFT-G test even after chemotherapy. PMID- 18399876 TI - High adenosine deaminase activity in the pleural effusion of a patient with Legionnaires' disease. AB - Although Legionnaires' disease (LD) is frequently accompanied by pleural effusion, the characteristics of pleural effusions in LD have not been well studied. Levels of adenosine deaminase (ADA) activity in pleural fluid >40 IU/L have a high sensitivity (81-100%) and specificity (83-100%) for tuberculosis. ADA activity in pleural effusions due to LD has not been previously reported. The case of a patient with LD complicated by a pleural effusion with high ADA activity is reported. In countries where the prevalence of tuberculosis is high and pleural fluid ADA activities are frequently measured, LD should be included in the differential diagnosis of an exudative pleural effusion with high ADA activity. PMID- 18399877 TI - Severe Legionella pneumonia successfully treated by independent lung ventilation with intrapulmonary percussive ventilation. AB - A case of severe Legionella pneumonia was successfully treated by independent lung ventilation (ILV) with intrapulmonary percussive ventilation (IPV). A 57 year-old man with lobar pneumonia was intubated and mechanically ventilated because of his deteriorating respiratory status. The diagnosis of Legionella pneumonia was made on the fourth day after admission and appropriate antibiotic therapy was commenced. On the fifth hospital day, ILV was commenced because the right unaffected lung was over-distended, his haemodynamic state was unstable and his left lung was producing copious amounts of purulent sputum. His right lung was ventilated and his left lung was treated with IPV owing to the existence of massive atelectasis. After treatment with antibiotics and ILV combined with IPV, his respiratory and haemodynamic status gradually improved. On the tenth day after admission, ILV was changed to conventional bilateral ventilation. The patient was extubated on the sixteenth hospital day and discharged from the intensive care unit 30 days after admission. The combination of ILV and IPV was therapeutically effective during the acute phase of unilateral severe Legionella pneumonia. PMID- 18399878 TI - Pseudallescheria boydii or Aspergillus fumigatus in a lady with an unresolving lung infiltrate, and a literature review. AB - A 53-year-old lady with blood-stained sputum and pleuritic pain had a lingular opacity on CXR which failed to resolve. A bronchial aspirate and transbronchial biopsy revealed features of bronchocentric granulomatosis with dichotomous branching hyphae suggestive of Aspergillus infection. However, subsequent fungal culture grew Pseudallescheria boydii. This case demonstrates the similarity of clinical and histological features caused by these two fungi. This appears to be the first reported case of pulmonary pseudallescheria with a bronchocentric granulomatous response. PMID- 18399879 TI - Thoracic splenosis: A diagnosis by history and imaging. AB - Thoracic splenosis (TS) is autoimplantation of ectopic splenic tissue in the thoracic cavity that occurs following splenic injury. The majority of cases of TS are asymptomatic and are diagnosed in the course of an evaluation of incidentally discovered pulmonary lesions. Some cases may be difficult to diagnose, especially if features suggesting TS are not recognized. This may lead to an extensive work up and unnecessary invasive diagnostic procedures including thoracotomy. This case report describes a 40-year-old man, who was diagnosed incidentally with TS, several years after the initial injury. Multiple, asymptomatic, left-sided pleural based lesions associated with a history of thoracoabdominal injury and splenectomy are the key points that should alert suspicion of TS, which can then simply be confirmed by a Tc-99m sulphur colloid radionuclide scan. Most patients are treated conservatively unless they are symptomatic. Physicians must recognize the key features suggesting a diagnosis of TS, order appropriate imaging studies and avoid unnecessary invasive diagnostic procedures. PMID- 18399880 TI - What do cross-national comparisons of personality traits tell us? The case of conscientiousness. AB - Much research contrasts self-reported personality traits across cultures. We submit that this enterprise is weakened by significant methodological problems (in particular, the reference-group effect) that undermine the validity of national averages of personality scores. In this study, behavioral and demographic predictors of conscientiousness were correlated with different cross national measures of conscientiousness based on self-reports, peer reports, and perceptions of national character. The predictors correlated strongly with perceptions of national character, but not with self-reports and peer reports. Country-level self- and peer-report measures of conscientiousness failed as markers of between-nation differences in personality. PMID- 18399881 TI - Naming practices and the acquisition of key biological concepts: evidence from English and Indonesian. AB - Children's acquisition of fundamental biological concepts (living thing, animal, plant) is shaped by the way these concepts are named. In English, but not Indonesian, the name "animal" is polysemous: One sense includes all animate objects, and the other excludes humans. Because names highlight object categories, if the same name ("animal") points to two different, hierarchically related biological concepts, children should have difficulty settling on the scope of that term and its close neighbors (e.g., "alive"). Experiments with 4- to 9-year-old English- and Indonesian-speaking children revealed that "alive" poses unique interpretive challenges, especially for English-speaking children. When asked to identify entities that are "alive," older Indonesian-speaking children selected both plants and animals, but their English-speaking counterparts tended to exclude plants, which suggests that they may have misaligned "alive" with one of the "animal" senses. This work underscores the importance of considering language and cultural factors in studying the acquisition of fundamental concepts about the biological world. PMID- 18399882 TI - Individual differences in insular sensitivity during loss anticipation predict avoidance learning. AB - The anterior insula has been implicated in both the experience and the anticipation of negative outcomes. Although individual differences in insular sensitivity have been associated with self-report measures of chronic anxiety, previous research has not examined whether individual differences in insular sensitivity predict learning to avoid aversive stimuli. In the present study, insular sensitivity was assessed as participants anticipated monetary losses while undergoing functional magnetic resonance imaging. We found that insular responsiveness to anticipated losses predicted participants' ability to learn to avoid losses (but not to approach gains) in a behavioral test several months later. These findings suggest that in addition to correlating with self-reported anxiety, heightened insular sensitivity may promote learning to avoid loss. PMID- 18399883 TI - Hedonic and instrumental motives in anger regulation. AB - What motivates individuals to regulate their emotions? One answer, which has been highlighted in emotion-regulation research, is that individuals are motivated by short-term hedonic goals (e.g., the motivation to feel pleasure). Another answer, however, is that individuals are motivated by instrumental goals (e.g., the motivation to perform certain behaviors). We suggest that both answers have merit. To demonstrate the role instrumental goals may play in emotion regulation, we pitted short-term hedonic motives and instrumental motives against each other, by testing whether individuals were motivated to experience a potentially useful, albeit unpleasant, emotion. We found that (a) individuals preferred activities that would increase their level of anger (but not their level of excitement) when they were anticipating confrontational, but not nonconfrontational, tasks and that (b) anger improved performance in a confrontational, but not a nonconfrontational, task. These findings support a functional view of emotion regulation, and demonstrate that in certain contexts, individuals may choose to experience emotions that are instrumental, despite short-term hedonic costs. PMID- 18399884 TI - Prescribed optimism: is it right to be wrong about the future? PMID- 18399885 TI - First impressions and last resorts: how listeners adjust to speaker variability. AB - Perceptual theories must explain how perceivers extract meaningful information from a continuously variable physical signal. In the case of speech, the puzzle is that little reliable acoustic invariance seems to exist. We tested the hypothesis that speech-perception processes recover invariants not about the signal, but rather about the source that produced the signal. Findings from two manipulations suggest that the system learns those properties of speech that result from idiosyncratic characteristics of the speaker; the same properties are not learned when they can be attributed to incidental factors. We also found evidence for how the system determines what is characteristic: In the absence of other information about the speaker, the system relies on episodic order, representing those properties present during early experience as characteristic of the speaker. This "first-impressions" bias can be overridden, however, when variation is an incidental consequence of a temporary state (a pen in the speaker's mouth), rather than characteristic of the speaker. PMID- 18399886 TI - The sunny side of fairness: preference for fairness activates reward circuitry (and disregarding unfairness activates self-control circuitry). AB - Little is known about the positive emotional impact of fairness or the process of resolving conflict between fairness and financial interests. In past research, fairness has covaried with monetary payoff, such that the mental processes underlying preference for fairness and those underlying preference for greater monetary outcome could not be distinguished. We examined self-reported happiness and neural responses to fair and unfair offers while controlling for monetary payoff. Compared with unfair offers of equal monetary value, fair offers led to higher happiness ratings and activation in several reward regions of the brain. Furthermore, the tendency to accept unfair proposals was associated with increased activity in right ventrolateral prefrontal cortex, a region involved in emotion regulation, and with decreased activity in the anterior insula, which has been implicated in negative affect. This work provides evidence that fairness is hedonically valued and that tolerating unfair treatment for material gain involves a pattern of activation resembling suppression of negative affect. PMID- 18399887 TI - Perceptual mechanisms that characterize gender differences in decoding women's sexual intent. AB - Men and women often disagree about the meaning of women's nonverbal cues, particularly those conveying dating-relevant information. Men perceive more sexual intent in women's behavior than women perceive or report intending to convey. Although this finding has been attributed to gender differences in the threshold for labeling ambiguous cues as sexual in nature, little research has been conducted to determine etiology. Using a model that differentiates perceptual sensitivity from decisional bias, we found no evidence that men have lenient thresholds for perceiving women's nonverbal behavior as indicating sexual interest. Rather, gender differences were captured by a relative perceptual insensitivity among men. Just as in previous studies, men were more likely than women to misperceive friendliness as sexual interest, but they also were quite likely to misperceive sexual interest as friendliness. The results point to the promise of computational models of perception in increasing the understanding of clinically relevant social processes. PMID- 18399888 TI - Personality dominance and preferential use of the vertical dimension of space: evidence from spatial attention paradigms. AB - Previous research has shown that dominant individuals frequently think in terms of dominance hierarchies, which typically invoke vertical metaphor (e.g., "upper" vs. "lower" class). Accordingly, we predicted that in spatial attention paradigms, such individuals would systematically favor the vertical dimension of space more than individuals low in dominance. This prediction was supported by two studies (total N= 96), which provided three tests involving two different spatial attention paradigms. In all cases, analyses controlling for speed of response to horizontal spatial probes revealed that more dominant individuals were faster than less dominant individuals to respond to probes along the vertical dimension of space. Such data support the metaphor-representation perspective, according to which people think in metaphoric terms, even in on-line processing tasks. These results have implications for understanding dominance and also indicate that conceptual metaphor is relevant to understanding the cognitive processing basis of personality. PMID- 18399889 TI - Beyond fear: rapid spatial orienting toward positive emotional stimuli. AB - There is much empirical evidence for modulation of attention by negative -- particularly fear-relevant -- emotional stimuli. This modulation is often explained in terms of a fear module. Appraisal theories of emotion posit a more general mechanism, predicting attention capture by stimuli that are relevant for the needs and goals of the organism, regardless of valence. To examine the brain activation patterns underlying attentional modulation, we recorded event-related potentials from 20 subjects performing a dot-probe task in which the cues were fear-inducing and nurturance-inducing stimuli (i.e., anger faces and baby faces). Highly similar validity modulation was found for the P1 time-locked to target onset, indicating early attentional capture by both positive and negative emotional stimuli. Topographic segmentation analysis and source localization indicate that the same amplification process is involved whether attention orienting is triggered by negative, fear-relevant stimuli or positive, nurturance relevant stimuli. These results confirm that biological relevance, and not exclusively fear, produces an automatic spatial orienting toward the location of a stimulus. PMID- 18399890 TI - Is happiness having what you want, wanting what you have, or both? AB - Rabbi Hyman Schachtel (1954) proposed that "happiness is not having what you want, but wanting what you have" (p. 37). In two studies, we tested Schachtel's maxim by asking participants whether or not they had and the extent to which they wanted each of 52 material items. To quantify how much people wanted what they had, we identified what they had and the extent to which they wanted those things. To quantify how much people had what they wanted, we identified how much they wanted and whether or not they had each item. Both variables accounted for unique variance in happiness. Moreover, the extent to which people wanted what they had partially mediated effects of gratitude and maximization on happiness, and the extent to which they had what they wanted partially mediated the effect of maximization. Results indicate that happiness is both wanting what you have and having what you want. PMID- 18399891 TI - Why it pays to get inside the head of your opponent: the differential effects of perspective taking and empathy in negotiations. AB - The current research explored whether two related yet distinct social competencies -- perspective taking (the cognitive capacity to consider the world from another individual's viewpoint) and empathy (the ability to connect emotionally with another individual) -- have differential effects in negotiations. Across three studies, using both individual difference measures and experimental manipulations, we found that perspective taking increased individuals' ability to discover hidden agreements and to both create and claim resources at the bargaining table. However, empathy did not prove nearly as advantageous and at times was detrimental to discovering a possible deal and achieving individual profit. These results held regardless of whether the interaction was a negotiation in which a prima facie solution was not possible or a multiple-issue negotiation that required discovering mutually beneficial trade offs. Although empathy is an essential tool in many aspects of social life, perspective taking appears to be a particularly critical ability in negotiations. PMID- 18399892 TI - The secret life of emotions. AB - The possibility of unconsciously evoked emotions is often denied because awareness of an emotion's cause is considered to be precisely what produces the emotion. However, we argue that because emotional responding is important for successful living, both global and specific emotional responses can be induced without awareness. The present research used quick and super-quick subliminal priming techniques, and cognitive, feelings, and behavioral measures, to test this hypothesis. Our results show that both global moods and specific emotions can be evoked without conscious awareness of their cause. PMID- 18399893 TI - The representation of simple ensemble visual features outside the focus of attention. AB - The representation of visual information inside the focus of attention is more precise than the representation of information outside the focus of attention. We found that the visual system can compensate for the cost of withdrawing attention by pooling noisy local features and computing summary statistics. The location of an individual object is a local feature, whereas the center of mass of several objects (centroid) is a summary feature representing the mean object location. Three experiments showed that withdrawing attention degraded the representation of individual positions more than the representation of the centroid. It appears that information outside the focus of attention can be represented at an abstract level that lacks local detail, but nevertheless carries a precise statistical summary of the scene. The term ensemble features refers to a broad class of statistical summary features that we propose collectively make up the representation of information outside the focus of attention. PMID- 18399894 TI - It takes two: the interpersonal nature of empathic accuracy. AB - Although current theories suggest that affective empathy (perceivers' experience of social targets' emotions) should contribute to empathic accuracy (perceivers' ability to accurately assess targets' emotions), extant research has failed to consistently demonstrate a correspondence between them. We reasoned that prior null findings may be attributable to a failure to account for the fundamentally interpersonal nature of empathy, and tested the prediction that empathic accuracy may depend on both targets' tendency to express emotion and perceivers' tendency to empathically share that emotion. Using a continuous affect-rating paradigm, we found that perceivers' trait affective empathy was unrelated to empathic accuracy unless targets' trait expressivity was taken into account: Perceivers' trait affective empathy predicted accuracy only for expressive targets. These data suggest that perceivers' self-reported affective empathy can indeed predict their empathic accuracy, but only when targets' expressivity allows their thoughts and feelings to be read. PMID- 18399895 TI - "In-group love" and "out-group hate" as motives for individual participation in intergroup conflict: a new game paradigm. AB - What motivates individual self-sacrificial behavior in intergroup conflicts? Is it the altruistic desire to help the in-group or the aggressive drive to hurt the out-group? This article introduces a new game paradigm, the intergroup prisoner's dilemma-maximizing difference (IPD-MD) game, designed specifically to distinguish between these two motives. The game involves two groups. Each group member is given a monetary endowment and can decide how much of it to contribute. Contribution can be made to either of two pools, one that benefits the in-group at a personal cost and another that, in addition, harms the out-group. An experiment demonstrated that contributions in the IPD-MD game are made almost exclusively to the cooperative, within-group pool. Moreover, preplay intragroup communication increases intragroup cooperation, but not intergroup competition. These results are compared with those observed in the intergroup prisoner's dilemma game, in which group members' contributions are restricted to the competitive, between-group pool. PMID- 18399896 TI - Synaesthesia, color terms, and color space: color claims came from color names in Beeli, Esslen, and Jancke (2007). PMID- 18399897 TI - Two year follow-up of immunological response in mite-allergic children treated with sublingual immunotherapy. Comparison with subcutaneous administration. AB - Although the efficacy of allergen-specific sublingual immunotherapy (SLIT) is now accepted, the underlying mechanisms remain elusive. Such mechanisms are better documented in the case of subcutaneous immunotherapy (SCIT). In order to understand the T-lymphocyte response in patients receiving SLIT, we compared children with respiratory disease monosensitized to Dermatophagoides pteronyssinus receiving SLIT or SCIT over a 2-yr period. Peripheral blood was obtained before beginning immunotherapy, and after 3 months, 1 yr and 2 yr. Total IgE, specific IgE and IgG4 to D. pteronyssinus were determined in serum. T-cell markers (CD3, CD4, CD8, CD25) and intracellular cytokine production (TNF-alpha, IL-2, IL-4 and IFN-gamma) were determined in peripheral blood mononuclear cells (PBMC) by flow cytometry. No differences between SCIT and SLIT were detected in the clinical variables or in the subjective evaluation. Although an increase in specific IgE and IgG4 was only detected in SCIT, a significant decrease in the specific IgE/IgG4 ratio was found in both groups. SCIT and SLIT experienced an increase in the CD4/CD8 ratio over time, but an increase in the CD4(+)CD25(+) and a decrease in the CD8(+)CD25(+) subsets were only found with SCIT. A slight shift from a Th2 to a Th1 pattern, measured by the IFN-gamma/IL-4 ratio, was only detected in the CD4 T cells with SCIT. A decrease in both groups was found in TNF alpha and IL-2 production over time. Children with respiratory allergic diseases receiving SCIT or SLIT had a different immunologic response in peripheral blood during treatment, though the clinical improvement was similar. Whether SLIT induces a mucosal protective response should be studied. PMID- 18399898 TI - Effect of nasal steroid treatment on airway inflammation determined by exhaled nitric oxide in allergic schoolchildren with perennial rhinitis and asthma. AB - Rhinitis is common in asthmatic schoolchildren who are allergic to animal dander and constantly and indirectly exposed to these allergens in their everyday environment. As a patho-physiological linkage between nasal and bronchial inflammation has been proposed to exist, the primary objective of this study was to determine whether nasal administration of mometasone furoate (MSNF) can reduce bronchial inflammation, as reflected in the level of exhaled nitric oxide (F(E)NO) in asthmatic schoolchildren with dander allergy and mild-to-moderate rhinitis. Forty such children were assigned randomly to be treated for 4 wk with MSNF or placebo, employing a double-blind procedure. F(E)NO was the primary end point measured and secondary end-points were nasal levels of NO, the concentration of eosinophilic cationic protein (ECP) in nasal lavage, the relative numbers of eosinophils in blood, forced expiratory volume in 1 s (FEV(1)), peak expiratory flow (PEF) and scoring of symptoms. There was no significant difference in the F(E)NO values of the treated and control groups at any time-point, whereas the nasal level of ECP was lower in the treated group compared with placebo (p = 0.05) on both days 7 and 28, and compared with baseline for the treated group (p = 0.06 on day 7, p = 0.02 on day 28). Furthermore, the mean blood eosinophil count decreased in the treated group, which also demonstrated lower scores for nasal symptoms compared with placebo, but neither of these differences were statistically significant. FEV(1), PEF and nasal levels of NO remained unchanged in both groups. Four weeks of nasal treatment with MSNF had no effect on bronchial inflammation, as reflected by exhaled NO, whereas signs of nasal and systemic eosinophil activation were reduced. Thus, nasal administration of a steroid as a strategy to reduce asthmatic inflammation remains questionable in mild-to-moderately severe cases of perennial rhinitis and asthma. PMID- 18399899 TI - Mycotic keratitis: an overview of diagnosis and therapy. AB - The increased incidence of fungal infections in the recent past has been attributed to the increase in the number of human immunodeficiency virus-positive and AIDS patients. Early diagnosis of mycoses in patients is crucial for prompt antifungal therapy. The yield of clinical examination in the diagnosis of keratomycosis is 63-83% and KOH mount is 91%. This still highlights the limitation of routine clinical examination and smear examination, which is not performing 100% efficiently. It is for these 37%, 17% and 9% of cases, every day advanced technologies are called for. Those who deal with patient care are aware of certainties and uncertainties of results of clinical examination. The best reported figures at specialized centres might not translate into clinical practice. Another factor to be kept in mind is that many patients who come after secondary and tertiary referrals are already treated with antibiotics, antivirals, steroids and sometimes even antifungals that distort the clinical picture completely. Further, one has to consider as well the cases caused by yeast-like fungi, which resemble bacterial keratitis. Confirmation of diagnosis, not only in case of mycotic keratitis but also for other diseases, to initiate prompt and accurate therapy would avoid unnecessary and indiscriminate use of steroids/antibacterials/antivirals and antifungals. PMID- 18399900 TI - Direct fluconazole susceptibility testing of positive Candida blood cultures by flow cytometry. AB - The standard methods for yeast susceptibility testing require 24-48 h of incubation. As there has been an increase in incidence of non-albicans Candida species, the clinician is very often wary of initiating therapy with fluconazole until a final susceptibility report is generated, especially when treating very sick patients. A rapid reliable susceptibility testing method would enable the clinician to prescribe fluconazole, thus avoiding more toxic or expensive therapy. To determine the feasibility of direct susceptibility testing of Candida species to fluconazole by a rapid flow cytometric method, 50 Candida strains were seeded into blood culture bottles and were tested for susceptibility to fluconazole directly from the bottles after their being flagged as positive by the blood culture instrument. Minimal inhibitory concentration (MIC) determined by fluorescent flow cytometry (FACS) showed excellent agreement to that determined by macrodilution. Following the seeding experiments, 30 true patient specimens were tested directly from positive blood cultures, and MIC determined by both methods showed excellent agreement. Antifungal susceptibility testing by FACS directly from positive blood culture bottles is a reliable, rapid method for susceptibility testing of Candida to fluconazole. The method allows same-day results, does not require subculture to agar media, and can greatly assist in the selection of appropriate antifungal therapy. PMID- 18399901 TI - Evaluation of Mueller-Hinton-agar as a simple medium for the germ tube production of Candida albicans and Candida dubliniensis. AB - Candida albicans is the most frequently isolated yeast species from clinical specimens. A classical rapid presumptive differentiation from non-albicans species is based on its ability to produce germ tubes after incubation in human serum. The only non-albicans Candida species producing germ tubes is Candida dubliniensis. In this study, we evaluated Mueller-Hinton-agar (MH-agar) as a medium for germ tube formation of C. albicans and C. dubliniensis. A total of 859 yeast isolates from stool samples, including 632 strains of C. albicans, 10 C. dubliniensis and 217 other yeast strains from 20 different species, were grown on Sabouraud glucose (2%) agar at 37 degrees C for 24-72 h. Species were identified by standard methods. For the germ tube test (GTT), an inoculum from a single colony was streaked onto a MH-agar plate and covered by a sterile coverslip. After incubation at 37 degrees C for 2 h, the MH plates were examined using a light microscope at x200. The GTT was positive in 578 of 632 C. albicans strains (sensitivity 91.5%), in six of 10 C. dubliniensis strains (sensitivity 60.0%), and in none of the other yeast strains. MH-agar is a suitable medium for the GTT and the presumptive identification of C. albicans. It is safer to use than human serum and is widely available in microbiology laboratories. PMID- 18399902 TI - Identification of clinically relevant yeast species by DNA sequence analysis of the D2 variable region of the 25-28S rRNA gene. AB - Clinically relevant yeasts are conventionally identified by a combination of phenotypic tests, which occasionally provide ambiguous results for atypical isolates or uncommon species. In this study, we evaluate a direct polymerase chain reaction-sequencing method, which exploits sequence divergence in the hypervariable D2 region of the large subunit of the 25-28S ribosomal RNA (rRNA) gene for identification of facultative pathogenic asco- and basidiomycota. A panel of 53 yeasts, including 40 clinical isolates and 13 reference strains representative of some clinically relevant taxa, was investigated by combining standard phenotypic tests with commercial identification systems (RapID, API 20C AUX), and results were compared with the taxonomic allocations inferred by D2 sequence analysis. Species-level resolution was achieved for almost all (52/53) strains by combining internet-based D2 sequence homology (BLAST and FASTA) searches in free-access synchronised databases with phylogenetic analysis. The phylogenetic information carried by the short D2 sequence substantiates a pattern of molecular evolution, which is similar to that inferred from analysis of the larger D1/D2 region, and consistent with previously published 25-28S rRNA phylogenetic architectures of facultative pathogenic yeast, including recently identified species. Inconsistency between conventional and molecular identification results was observed for 11/53 strains, likely on account of the ambiguous interpretation of phenotypic tests. PMID- 18399903 TI - Innate and adaptive immunity in patients with autoimmune polyendocrinopathy candidiasis-ectodermal dystrophy. AB - Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is an autosomal recessive syndrome characterised by chronic mucocutaneous candidiasis (CMC) and multiple endocrine failures. While the spectrum and modalities of autoimmunity are the main objects of current research into APECED, unequivocal data on the efficiency of immune responses to infectious agents are still elusive. The in vitro ability of monocytes and polymorphonuclear leucocytes to phagocytise and kill bacteria and fungi, and the degree of activation of lymphocytes cultured with mitogens and Candida albicans were investigated by flow cytometry in 11 APECED patients and healthy subjects. In addition, a comparison of gamma-globulin and immunoglobulin (Ig) concentrations was performed, and a correlation was sought between oral fungal load and the anti-Candida antibody titre. No difference between APECED patients and healthy subjects was observed in the phagocyte function, although the patients had a larger number of monocytes. Similarly, cultured lymphocytes were equally activated in the two groups. The concentration of gamma-globulins was higher among APECED patients, and anti Candida IgM and IgG correlated with current and past oral candidiasis respectively. APECED patients have efficient innate and adaptive immune responses against exogenous stimuli, and currently, the mechanisms of mucocutaneous anergy leading to the high prevalence of CMC in this syndrome remain to be elucidated. PMID- 18399904 TI - Clinical assessment of postinfection, postcontact and postvaccination immunity manifestation after experimental inoculation of calves with Trichophyton verrucosum culture. AB - Immunity against ringworm has been well examined, but the knowledge of its development after repeated infection is limited. Therefore, the immunity developed after a single or double contact with various doses of Trichophyton verrucosum strain was investigated in calves which had or had not shown any signs of ringworm after contact with dermatophyte. In total six individual experiments were performed on 117 calves. Experimental infection with the virulent T. verrucosum strain was used to investigate the animals' immunity. Calves showing severe trichophytosis after application of high doses of T. verrucosum culture were immune to reinfection by equally high doses of this dermatophyte. A similar protection level was reached after intramuscular revaccination or after recovery from trichophytosis with moderate course induced by two inoculations of low doses. If moderate course of illness had been induced by same single dose, the immunity was lower. Animals displaying no signs of trichophytosis after inoculation with the low doses developed insufficient immunity. There was no immunity against reinfection in animals having previous contact with subinfectious doses through undamaged skin. The results obtained suggest that the level of postinfection immunity depends on the disease extent and the frequency of administration of the infectious agent. The development of postcontact immunity cannot be excluded in calves. PMID- 18399905 TI - Management and follow-up survey of Trichophyton tonsurans infection in a university judo club. AB - The prevalence of Trichophyton tonsurans infection of the scalp in members of a university judo club (combat sport) was investigated over a 3.5-year period using a questionnaire survey and an assay based on fungal culture by the hairbrush method. In November 2002, 11 (35%) of 31 athletes were found to be positive for T. tonsurans infection by the hairbrush method and provided treatment with oral and topical antifungal agents according to a prescribed protocol. All the infected subjects became culture-negative following this treatment. We continued to conduct screening examinations every year in the month of April, when new university enrolment occurs. During three-and-a-half years of follow-up, there have been no outbreaks of the infection among the members of the university judo club. There were some positive culture results among the newly enrolled students, but these cases also became culture-negative with treatment. No re-infection has been noted after graduation among the club members who had been educated about and treated for the infection. Our findings indicate that the spread of T. tonsurans infection in sports clubs can be controlled by regular mass screening examination, therapy and measures at regular intervals to prevent the infection. PMID- 18399906 TI - Nosocomial candidaemia in children: results of a 9-year study. AB - The aim of this study was to determine changes in the incidence of nosocomial candidaemia and to evaluate the risk factors, demographic features, treatment and clinical outcome associated with candidaemia in a Turkish tertiary care paediatric unit within a 9-year period. The data of children who were diagnosed as nosocomial candidaemia, were examined in this study. Between January 1997 and December 2005, a total of 102 nosocomial candidaemia episodes were identified in 102 patients. The rate of nosocomial candidaemia in our clinic increased from 3.2 cases per 1000 admissions in 1997-1999, to 5.5 per 1000 admissions in 2000-2002 and to 6.9 per 1000 admissions in 2003-2005 (P = 0.003). The species most frequently causing candidaemia were Candida albicans (39.2%), Candida parapsilosis (21.6%) and Candida tropicalis (15.7%). The mortality of C. albicans (37.5%), was significantly higher than the mortality of non-albicans species (17.7%) (P = 0.04). Independent risk factors associated with candidaemia-related deaths by logistic regression analysis were disseminated candidiasis (odds ratio, 5.7; P = 0.01), paediatric intensive care unit stay (odds ratio, 8.1; P = 0.001), prolonged antibiotics therapy (odds ratio, 5.2; P = 0.014), use of total parenteral nutrition (odds ratio, 4.4; P = 0.038) and mechanical ventilation (odds ratio, 4.9; P = 0.01). The rate of nosocomial candidaemia in our clinic increased >2-fold during the study period. PMID- 18399907 TI - Current scenario of cryptococcosis and antifungal susceptibility pattern in India: a cause for reappraisal. AB - This study analysed the spectrum, antifungal susceptibility pattern, clinical course and molecular epidemiology of cryptococcosis. Four hundred and thirty-nine samples obtained from 378 meningitis patients were processed by standard procedures. Minimum inhibitory concentration (MIC) of fluconazole and amphotericin B for the isolates was tested by broth micro dilution and by E-strip method. Molecular analysis by random amplified polymorphic DNA-PCR of eight isolates was performed using M13 primer. Cryptococcosis was diagnosed in 35 patients [HIV-1 seropositive (19) and apparently immunocompetent (16)]. Cryptococcus neoformans var. neoformans (serotype A and D) was the predominant isolate on phenotypic identification. Three C. neoformans var. gattii were isolated from HIV-1 seropositive (2) and apparently immunocompetent (1) patients. MIC 90 for amphotericin B and fluconazole were 1 and 8 mug ml(-1) respectively. On RAPD-PCR, less diversity was seen among Indian isolates. AIDS remains the single most important risk factor for cryptococcosis. Rising MIC of the available induction and maintenance drugs is of grave concern. The DNA typing technique showed less diversity among Indian strains. Routine surveillance and application of molecular typing methods are crucial to know the baseline and existing pattern of cryptococcosis. PMID- 18399908 TI - Lack of evidence of endosymbiotic toxin-producing bacteria in clinical Rhizopus isolates. AB - Infections by Rhizopus spp. account for about 90% of zygomycoses, many of which are lethal in immunocompromised patients. We recently noted that several strains of Rhizopus microsporus harbour rare bacterial endosymbionts (Burkholderia sp.) for the production of 'mycotoxins', which might play a role as virulence factors in human Rhizopus infections. In this study eight clinical Rhizopus spp. isolates have been investigated for the presence of toxin-producing bacterial endosymbionts. By metabolomic data, PCR targeting bacterial 16S rDNA and microscopic investigations with fluorescence dyes we provide three lines of evidence showing that the fungal strains are not associated with endofungal bacteria. Consequently, toxin-producing bacteria are not essential for Rhizopus infections and the development of zygomycoses in humans. PMID- 18399909 TI - Successful treatment of disseminated Geotrichum capitatum infection with a combination of caspofungin and voriconazole in an immunocompromised patient. AB - Disseminated Geotrichum capitatum infection is uncommon, and has been reported exclusively in immunocompromised patients. The prognosis is poor with a mortality rate of approximately 50-75%. We report a case of disseminated G. capitatum infection in a severely neutropenic patient who was receiving chemotherapy for acute myeloblastic leukaemia. G. capitatum was isolated from blood cultures, skin lesions, bronchoalveolar lavage fluid, throat swabs and stools. The infection was successfully cured with a combination of voriconazole and caspofungin. PMID- 18399910 TI - Abstracts of the 14th Annual Conference of the British HIV Association. April 23 25, 2008. Belfast, United Kingdom. PMID- 18399911 TI - Abstracts of the 18th European Congress of Clinical Microbiology and Infectious Diseases. Barcelona, Spain. April 19-22, 2008. PMID- 18399912 TI - Aire deficiency causes increased susceptibility to streptozotocin-induced murine type 1 diabetes. AB - Aire-deficient mice are a model of the human monogenic disorder autoimmune polyendocrine syndrome type I (APS I) characterized by a progressive autoimmune destruction of multiple endocrine glands such as the adrenal cortex, the parathyroids and the beta-cells of the pancreas. The disease is caused by mutations in the autoimmune regulator (AIRE) gene, a putative transcription factor expressed in thymic medullary epithelial cells and in antigen-presenting cells of the myeloid lineage in peripheral lymphoid organs. As Aire(-/-) mice do not spontaneously develop endocrinopathies, we wanted to evaluate the autoimmune multiple low-dose streptozotocin (MLDSTZ) diabetes model in Aire(-/-) mice. Surprisingly, Aire heterozygote mice were most susceptible to MLDSTZ-induced diabetes, whereas Aire(-/-) mice displayed an intermediate sensitivity to diabetes. Furthermore, Aire(-/-) macrophages produced higher levels of TNF-alpha and lower levels of IL-10 following streptozotocin stimulation, and Aire(-/-) mice developed a higher frequency of islet cells autoantibodies as a sign of increased activation. However, the number of islet infiltrating F4/80(+) Aire(-/ ) macrophages was significantly decreased which was attributed to an increased susceptibility to streptozotocin cytotoxicity of Aire(-/-) macrophages. In conclusion, Aire(-/-) macrophages display an increased activation after STZ stimuli, but suffer from increased susceptibility to STZ cytotoxicity. These results support an important function of Aire in the control of peripheral tolerance through myeloid antigen-presenting cells. PMID- 18399913 TI - Lewis-acid-catalyzed photodimerization of coumarins and N-methyl-2-quinolone. AB - The BF(3)-catalyzed photodimerization of parent coumarin (1), three 6 alkylcoumarins (2-4) and N-methyl-2-quinolone (5) in dichloromethane was studied by time-resolved UV-vis spectroscopy. The lowest triplet state properties in the absence and presence of BF(3) were outlined, in particular the effect of self quenching which initiates dimerization. The quantum yield of intersystem crossing (Phi(isc)) of 1-4 increases with BF(3) concentration, approaching Phi(isc) = 0.3. Phi(isc) and the relative quantum yield of dimerization go along, thereby favoring an overall triplet mechanism in both the direct and BF(3)-catalyzed photodimerization. The product ratio of 5 changes strongly with the BF(3) concentration from 100%anti-hh for the free quinolone to 100%syn-ht for the 1:1 complex. PMID- 18399914 TI - Rhodopsin regeneration is accelerated via noncovalent 11-cis retinal-opsin complex--a role of retinal binding pocket of opsin. AB - The regeneration of bovine rhodopsin from its apoprotein opsin and the prosthetic group 11-cis retinal involves the formation of a retinylidene Schiff base with the epsilon-amino group of the active lysine residue of opsin. The pH dependence of a Schiff base formation in solution follows a typical bell-shaped profile because of the pH dependence of the formation and the following dehydration of a 1-aminoethanol intermediate. Unexpectedly, however, we find that the formation of rhodopsin from 11-cis retinal and opsin does not depend on pH over a wide pH range. These results are interpreted by the Matsumoto and Yoshizawa (Nature 258 [1975] 523) model of rhodopsin regeneration in which the 11-cis retinal chromophore binds first to opsin through the beta-ionone ring, followed by the slow formation of the retinylidene Schiff base in a restricted space. We find the second-order rate constant of the rhodopsin formation is 6100+/-300 mol(-1) s(-1) at 25 degrees C over the pH range 5-10. The second-order rate constant is much greater than that of a model Schiff base in solution by a factor of more than 10(7). A previous report by Pajares and Rando (J Biol Chem 264 [1989] 6804) suggests that the lysyl epsilon-NH(2) group of opsin is protonated when the beta ionone ring binding site is unoccupied. The acceleration of the Schiff base formation in rhodopsin is explained by stabilization of the deprotonated form of the lysyl epsilon-NH(2) group which might be induced when the beta-ionone ring binding site is occupied through the noncovalent binding of 11-cis retinal to opsin at the initial stage of rhodopsin regeneration, followed by the proximity and orientation effect rendered by the formation of noncovalent 11-cis retinal opsin complex. PMID- 18399915 TI - Chromophore interaction in xanthorhodopsin--retinal dependence of salinixanthin binding. AB - Xanthorhodopsin is a light-driven proton pump in the extremely halophilic bacterium Salinibacter ruber. Its unique feature is that besides retinal it has a carotenoid, salinixanthin, with a light harvesting function. Tight and specific binding of the carotenoid antenna is controlled by binding of the retinal. Addition of all-trans retinal to xanthorhodopsin bleached with hydroxylamine restores not only the retinal chromophore absorption band, but causes sharpening of the salinixanthin bands reflecting its rigid binding by the protein. In this report we examine the correlation of the changes in the two chromophores during bleaching and reconstitution with native all-trans retinal, artificial retinal analogs and retinol. Bleaching and reconstitution both appear to be multistage processes. The carotenoid absorption changes during bleaching occurred not only upon hydrolysis of the Schiff base but continued while the retinal was leaving its binding site. In the case of reconstitution, the 13-desmethyl analog formed the protonated Schiff base slower than retinal, and provided the opportunity to observe changes in carotenoid binding at various stages. The characteristic sharpening of the carotenoid bands, indicative of its reduced conformational heterogeneity in the binding site, occurs when the retinal occupies the binding site but the covalent bond to Lys-240 via a Schiff base is not yet formed. This is confirmed by the results for retinol reconstitution, where the Schiff base does not form but the carotenoid exhibits its characteristic spectral change from the binding. PMID- 18399916 TI - Low-temperature spectroscopy of Met100Ala mutant of photoactive yellow protein. AB - The trans-to-cis photoisomerization of the p-coumaroyl chromophore of photoactive yellow protein (PYP) triggers the photocycle. Met100, which is located in the vicinity of the chromophore, is a key residue for the cis-to-trans back isomerization of the chromophore, which is a rate-determining reaction of the PYP photocycle. Here we characterized the photocycle of the Met100Ala mutant of PYP (M100A) by low temperature UV-visible spectroscopy. Irradiation of M100A at 80 K yielded a 380 nm species (M100A(BL)), while the corresponding intermediate of wild type (WT; PYP(BL)) is formed above 90 K. The amounts of redshifted intermediates produced from M100A (M100A(B') and M100A(L)) were substantially less than those from WT. While the near-UV intermediate (PYP(M)) is not formed from WT in glycerol samples at low temperature, M100A(M) was clearly observed above 190 K. These alterations of the photocycle of M100A were explained by the shift in the equilibrium between the intermediates. The carbonyl oxygen of the thioester linkage of the cis-chromophore in the photocycle intermediates is close to the phenyl ring of Phe96 (<3.5 A), which would be displaced by the mutation of Met100. These findings imply that the interaction between chromophore and amino acid residues near Met100 is altered during the early stage of the PYP photocycle. PMID- 18399917 TI - Identification of six new photoactive yellow proteins--diversity and structure function relationships in a bacterial blue light photoreceptor. AB - Photoactive yellow proteins (PYP) are bacterial photoreceptors with a Per-Arnt Sim (PAS) domain fold. We report the identification of six new PYPs, thus nearly doubling the size of this protein family. This extends the taxonomic diversity of PYP-containing bacteria from photosynthetic to nonphotosynthetic bacteria, from aquatic to soil-dwelling organisms, and from Proteobacteria to Salinibacter ruber from the phylum Bacteriodetes. The new PYPs greatly increase the sequence diversity of the PYP family, reducing the most prevalent pair-wise identity from 45% to 25%. Sequence alignments and analysis indicate that all 14 PYPs share a common structure with 13 highly conserved residues that form the chromophore binding pocket. Nevertheless, the functional properties of the PYPs vary greatly- the absorbance maximum extends from 432 to 465 nm, the pK(a) of the chromophore varies from pH 2.8 to 10.2, and the lifetime of the presumed PYP signaling state ranges from 1 ms to 1 h. Thus, the PYP family offers an excellent opportunity to investigate how functional properties are tuned over a wide range, while maintaining the same overall protein structural fold. We discuss the implications of these results for structure-function relationships in the PYP family. PMID- 18399918 TI - Quantitation of the effect of hydroxylamine on rhodopsin palmitylation. AB - Rhodopsin (the photosensitive rod visual pigment) has been a model for photobiologic studies of the opsins as well as a structural model for G-protein coupled receptors. The two palmitate groups attached to cysteines 322 and 323 are thought to serve as membrane anchors for the rhodopsin C-terminus, but the absence of the palmitates does not alter membrane localization. However, removal of the palmitates affects rhodopsin function. Therefore, it is important to quantitate the stability of rhodopsin palmitates to hydroxylamine, which is a widely utilized reagent in biochemical preparations of the apoprotein. We have developed a mass spectrometric method to quantitate the resulting opsin palmitylation. Our data show that both of the bovine rhodopsin palmitates are labile to hydroxylamine, with significant depalmitylation occurring at concentrations of >or=100 mM, with an EC(50) of 220 mM L(-1). The palmitate at position 322 is the more stable to hydroxylamine. Samples prepared in the presence of >50 mM should therefore be considered to be at least partially depalmitylated and the results interpreted accordingly. PMID- 18399919 TI - Photo-induced reactive oxygen species generation by different water-soluble fullerenes (C) and their cytotoxicity in human keratinocytes. AB - In this study we report the phototoxicity toward HaCaT keratinocytes that results from the photogeneration of superoxide and singlet oxygen ((1)O(2)) by four different "water-soluble" fullerene (C(60)) preparations-monomeric (gamma CyD)(2)/C(60) (gamma-cyclodextrin bicapped C(60)) and three aggregated forms THF/nC(60) (prepared by solvent exchange from THF solution); Son/nC(60) (prepared by sonication of a toluene/water mixture); and gamma-CyD/nC(60) (prepared by heating the [gamma-CyD](2)/C(60) aqueous solution). Our results demonstrate that all four C(60) preparations photogenerate (1)O(2) efficiently. However, the properties of C(60)-generated (1)O(2), including its availability for reactions in solution, are markedly different for the monomeric and aggregated forms. (1)O(2) produced by monomeric (gamma-CyD)(2)/C(60) is quenchable by NaN(3) and its quantum yield in D(2)O, which is only weakly dependent on oxygen concentration, is as high as C(60) in toluene. In contrast, (1)O(2) generated from aggregated C(60) is not quenchable by NaN(3), exhibits a solvent-independent short-lived lifetime (ca 2.9 micros), is highly sensitive to oxygen concentration while its phosphorescence is redshifted. All these features indicate that (1)O(2) is sequestered inside the C(60) aggregates, which may explain why these preparations were not phototoxic toward HaCaT cells. Electron paramagnetic resonance studies demonstrated the generation of the C(60) anion radical (C(60)) when (gamma-CyD)(2)/C(60) was irradiated (lambda > 300 nm) in the presence of a reducing agent (NADH); spin trapping experiments (lambda > 400 nm) with 5,5 dimethyl-1-pyrroline N-oxide clearly showed the generation of superoxide resulting from the reaction of C(60) with oxygen. In vitro tests with HaCaT keratinocytes provided evidence that (gamma-CyD)(2)/C(60) phototoxicity is mainly mediated by (1)O(2) (Type II mechanism) with only a minor contribution from free radicals (Type I mechanism). PMID- 18399920 TI - Thermal recovery of iodopsin from photobleaching intermediates. AB - The chloride effect on the photobleaching process of iodopsin, a chicken red sensitive cone visual pigment, was studied in detail by time-resolved low temperature spectroscopy at -40 degrees C to -10 degrees C. Decay-associated difference spectra obtained by kinetic analysis using the singular value decomposition method were composed of spectra of BL-iodopsin, lumiiodopsin, metaiodopsin I, metaiodopsin II and metaiodopsin III, essentially identical to those at room temperature. In each conversion step however, iodopsin was partially regenerated, which is not observed in the bleaching process for other visual pigments or iodopsin at room temperature. Moreover, iodopsin was slowly regenerated from the bleached species. The reverse reactions were completely suppressed by substitution of lyotropic NO(3)(-) for Cl(-), suggesting that Cl(-) binding to iodopsin interferes with light-induced cis-trans isomerization of the chromophore. It is likely that the water molecule hydrating Cl(-) forms the additional hydrogen bond(s), by which the protein conformational change necessary to release this steric hindrance becomes enthalpic. As progress of the bleaching process is a consequence of protein conformational change, it is suppressed at low temperatures, resulting in thermal back-isomerization. PMID- 18399921 TI - Surface elastic properties of human retinal pigment epithelium melanosomes. AB - Atomic force microscope (AFM) imaging and nanoindentation measurements in water were used to probe the mechanical properties of retinal pigment epithelium melanosomes isolated from 14-year-old and 76-year-old donors. Topographic imaging reveals surface roughness similar to previous measurements on dry melanosomes. Force-indentation measurements show different types of responses that were catalogued into four different categories. In these measurements no permanent surface damage of melanosomes was observed as revealed by imaging before and after indentation measurements. The indentation measurements that exhibited nearly elastic responses were used to determine the Young's modulus of melanosomes. The average Young's modulus values are similar for 14-year-old and 76-year-old melanosomes with a somewhat narrower distribution for the 14-year-old sample. These elastic modulus values are considerably higher than the modulus of organelles with cytoplasm (<1 MPa) and approaching values of the modulus of protein crystals (approximately 100 MPa) indicating rather high packing density of biologic material in melanosomes. The width of the Young's modulus distributions is considerable spanning from few megapascals to few tens of megapascals indicating large heterogeneity in the structure. A fraction of the force curves cannot be described by the homogeneous elastic sample model; these force curves are consistent with approximately 10 nm structural heterogeneity in melanosomes. The approach-withdraw hysteresis indicates a significant viscoelasticity, particularly in the samples from the 14-year-old sample. Adhesion of the AFM probe was detected on approximately 3% and approximately 20% of the surface of 14-year-old and 76-year-old samples, respectively. In light of previous studies on these same melanosomes using photoelectron emission microscopy, this adhesion is attributed to the presence of lipofuscin on the surface of the melanosomes. This suggestion indicates that part of the difference in photochemical properties between the old and young melanosomes originates from surface lipofuscin. PMID- 18399922 TI - Trends in UV irradiance of tanning devices in Norway: 1983-2005. AB - Indoor tanning increases skin cancer risk, but the importance of different parts of the UV spectrum is unclear. We assessed irradiance of tanning devices in Norway for the period 1983-2005. Since 1983, all tanning models needed approval before being sold or used. UV Type 3 limits were valid from late 1992 (<0.15 W m( 2) for CIE-weighted, i.e. erythemally weighted, short and long wave irradiances). We analyzed data from 90% of the approved tanning models (n = 446 models) and two large inspection surveys in 1998/1999 and 2003 (n = 1,341 tanning devices). Mean CIE-weighted short wave irradiance of approved models increased from 0.050 W m( 2) (95% confidence interval [CI] 0.045-0.055) in 1983-1992 to 0.101 W m(-2) (95% CI 0.098-0.105) in 1993-2005, and mean long wave from 0.091 W m(-2) (95% CI 0.088 0.095) to 0.112 W m(-2) (95% CI 0.109-0.115), respectively. Inspection surveys revealed short wave irradiances much higher than that approved. In 1998-1999, only 28% (293/1034) of the devices were equipped with correct sunlamps and only 1 out of 130 inspected establishments fulfilled all requirements. In 2003, corresponding numbers were 59% (180/307) of devices and 2 out of 52 establishments. Mean short and long wave irradiances of the inspected tanning devices in 2003 were 1.5 and 3.5 times, respectively, higher than the irradiance of natural summer sun in Oslo. In conclusion, the short wave irradiance has increased in indoor tanning devices in Norway over the last 20 years. Due to the high long wave irradiance throughout this period, the percentage of short wave irradiance was much lower than for natural sun. PMID- 18399923 TI - Transient vibronic structure in ultrafast fluorescence spectra of photoactive yellow protein. AB - The ultrafast photo-induced dynamics of wild-type photoactive yellow protein and its site-directed mutant of E46Q in aqueous solution was studied at room temperature by femtosecond fluorescence spectroscopy using the optical Kerr-gate method. The vibronic structure appears, depending on the excitation photon energy, in the time-resolved fluorescence spectra just after photoexcitation, which winds with time and disappears on a time scale of sub-picoseconds. This result indicates that the wavepacket is localized in the electronic excited state followed by dumped oscillations and broadening, and also that the initial condition of the wavepacket prepared depending on the excitation photon energy affects much the following ultrafast dynamics in the electronic excited state. PMID- 18399924 TI - Quantification of naturally occurring pyrrole acids in melanosomes. AB - Three naturally occurring pyrrole acids were found in Sepia, human black hair, and bovine choroid and iris melanosomes using high-performance liquid chromatography and mass spectrometry--pyrrole-2,3-dicarboxylic acid (PDCA), pyrrole-2,3,5-tricarboxylic acid (PTCA) and pyrrole-2,3,4,5-tetracarboxylic acid (PTeCA). PDCA and PTCA are common markers quantified from oxidative degradation of eumelanins. Using standards, the amounts of naturally occurring PDCA and PTCA were determined and compared to those obtained following peroxide oxidation of the same samples. Because the naturally occurring acids are water soluble, these results indicate that care must be exercised when comparing PDCA and PTCA yields from the degradation analyses of melanins isolated and prepared by different methods. This work also establishes that PTeCA is a naturally occurring pyrrole acid in melanosomes. PMID- 18399925 TI - Comparable photoreactivity of hair melanosomes, eu- and pheomelanins at low concentrations: low melanin a risk factor for UVA damage and melanoma? AB - Melanin is known to be photoreactive and photoprotective, but its function in skin in vivo is still debated. Data is lacking of the effects of UVA irradiation on human skin melanosomes of different pigmentation, which have not been extensively degraded by isolation procedures. We have shown previously that melanosomes isolated from human oriental and black cat hair, and synthetic eumelanins, are photoreactive producing superoxide at low concentrations when exposed to UVA irradiation comparable to UK levels of sunlight. Here we investigated the UVA-irradiation of melanosomes, isolated from different colored human hair samples, using electron spin resonance spectroscopy and spin trapping. Comparable irradiation of synthetic pheomelanins synthesized from L-dopa and L cysteine was also studied. An alkali method (5 min NaOH at 90 degrees C) could be used to isolate oriental hair melanosomes but was not suitable for auburn and blonde hair. Dithiothreitol and proteinase K resulted in melanin release from possible over-digestion of melanosomes; however, dithiothreitol and papain resulted in no melanin release and good melanosome yields with separation from residual keratin for brown, auburn and blonde hair. Melanosomes isolated by the latter method and synthetic pheomelanins were similar in UVA-photoreactivity at low concentrations, independent of hair color, and broadly comparable to synthetic melanins. Melanosome concentration at constant fluence may be more significant with respect to photodamage and UVA photocarcinogenesis (melanoma) via superoxide radical production than pigment type. PMID- 18399926 TI - Introduction to the Symposium-in-Print on Melanins. PMID- 18399927 TI - Ascorbate enhances photogeneration of hydrogen peroxide mediated by the iris melanin. AB - The iris in the human eye is exposed to UV and visible light transmitted by the cornea. This pigmented tissue is bathed with the aqueous humor (AqH), which contains high concentration of ascorbate. It has been postulated that the presence of ascorbate in AqH can contribute to increased photoproduction of H2O2 mediated by the iris melanin. In this study, we monitored generation of H2O2 induced by UV-VIS irradiation of bovine irides, bovine and human iris homogenates and iris melanin. Our data show that exogenous ascorbate significantly amplified the rate of H2O2 photoformation in all melanotic samples. Deactivation of endogenous catalase with NaN3 in bovine irides increased the level of the accumulated H2O2 in the bathing solution following sample irradiation. Photoformation of H2O2 in samples with exogenous ascorbate was accompanied by its photo-oxidation. Both photoprocesses exhibited significant wavelength dependence. EPR spectroscopy measurements showed that ascorbyl radical is an intermediate product of the ascorbate photo-oxidation. The photoproduction of H2O2 and photo oxidation of ascorbate appear to be stoichiometric processes. No significant differences in the photoreactivity of iridial melanin from donors of different age and iris color was found. We postulate that also in vivo ascorbate increases the rate of iris melanin-mediated photoformation of H2O2 and its steady state concentration in AqH. PMID- 18399928 TI - Prompt assessment of WST11-VTP outcome using luciferase transfected tumors enables second treatment and increase in overall therapeutic rate. AB - This study hypothesized that success rate assessment of vascular targeted photodynamic therapy (VTP) of solid tumors 24 h post-treatment may allow prompt administration of a second treatment in case of failure, increasing the overall success rate. Here, we show that treatment of luciferase transfected CT26-luc mouse colon carcinoma tumors in BALB/c mice by VTP with WST11 (a Pd bacteriochlorophyll-based photosensitizer) allows fast assessment of treatment success 24 h post-treatment, using bioluminescence imaging (BLI). WST11-VTP was found to abolish luciferin bioluminescence in the treated tumors resulting in two types of responses. One, comprising 75% of the mice, signified successful outcome, presenting neither BLI signal nor tumor regrowth (24 h-90 days post VTP). The second (the remaining 25% of the mice) signified treatment failure, presenting various levels of BLI signal with subsequent tumor regrowth (24 h-90 days). Consequently, the mice that failed the first treatment were treated again. We show that treatment success rate in both VTP sessions was identical and that the cumulative success rate of the treatment increased from 75% to over 90%. These results therefore, present a fast method of assessing VTP outcome and support the feasibility of successive multiple treatments with these sensitizers in the clinical arena. The presented methodology can also be helpful in future preclinical studies, and expedite the development of VTP drugs. PMID- 18399929 TI - Experimental measurements of leaf carbon isotope discrimination and gas exchange in the progenies of Plantago depressa and Setaria viridis collected from a wide altitudinal range. AB - Significant correlations between leaf carbon isotope discrimination (Delta) and altitude and between gas exchange and altitude have been reported in previous studies, raising the question of whether the altitudinal variations in discrimination and gas exchange can be attributed to genetic differences among populations from different altitudes. Studies that focus on in situ analysis cannot distinguish the effects of genetic variation from environmental variation. This article describes an experiment in which seeds of Plantago depressa (C3 species) and Setaria viridis (C4 species) collected from a wide altitudinal range were grown in the same environment. Carbon isotopic ratios (delta(13)C) and gas exchange of the seedlings were measured. The progenies of P. depressa and S. viridis no longer display any significant Delta decreases with the altitude of origin as seen in situ. Furthermore, photosynthetic rate, stomatal conductance, the ratio of intercellular to ambient CO(2) and intrinsic water use efficiency for P. depressa and S. viridis grown in the greenhouse are also not significantly related to the altitude of origin. The observations suggest that altitudinal variations in Delta and gas exchange are not because of genotypic differences, independent of photosynthetic type. PMID- 18399930 TI - Influence of intra-shoot trophic competition on shoot development in two grapevine cultivars (Vitis vinifera). AB - The effect of trophic competition between vegetative sources and reproductive sinks on grapevine (Vitis vinifera L.) shoot development was analyzed. Two international cultivars (Grenache N and Syrah) grown in pots, which were well watered, were studied. A large range of trophic competition levels was obtained by modifying the cluster loads per plant. An analytical breakdown of the branching system was used to analyze the effects of trophic competition. Phytomer production on the primary axis and the probability and timing of axillary budburst were not affected by trophic competition. However, the duration of development and leaf production rate for secondary axes were both significantly affected. The impact of trophic competition differed within the P0-P1-P2 architectural module, locally within the shoot and between cultivars. Trophic competition reduced the organogenesis of secondary axes most strongly close to clusters, on P1-P2 phytomers and in Grenache N. Based on these results, a modeling approach simulating sink strength variation and the local effects of sink proximity would be more relevant than a model considering only development as a function of thermal time or the global distribution of available biomass. PMID- 18399931 TI - Diagnostic screening of MODY2/GCK mutations in the Norwegian MODY Registry. AB - BACKGROUND: Maturity-onset diabetes of the young, type 2 (MODY2) is caused by mutations in the glucokinase gene (GCK). The aim of our study was to determine the prevalence of GCK mutations in the Norwegian MODY Registry and to delineate the clinical phenotype of identified GCK mutation carriers. METHODS: We screened 122 probands referred to the MODY Registry for mutations in GCK and studied extended families with MODY2. RESULTS: We found 2 novel (S76Y and N231S) and 13 previously reported (V62A, G72R, L146R, R191W, A208T, M210K, Y215X, M235T, R275C, E339G, R377C, S453L, and IVS5+1G>C) GCK mutations in 23 probands and in their 33 family members. The prevalence of MODY2 was 12% in the Norwegian MODY Registry. The subjects with GCK mutations had features of mild diabetes. Yet, 15 of 56 MODY2 subjects were treated with oral drugs or insulin. Three subjects had retinopathy and one had macrovascular disease. Also, a limited number of cases had elevated fasting serum triglyceride values. Moreover, two GCK mutation carriers were diagnosed with type 1 diabetes. CONCLUSIONS: According to our diagnostic screening of GCK in the MODY Registry, MODY2 is less prevalent than MODY3 in Norway but is likely to be underreported. Recognizing MODY2 in diabetic patients is important in order to prevent overtreatment. Finally, our study demonstrates the co-occurrence of MODY2 in families with type 1 or type 2 diabetes. PMID- 18399932 TI - NtGNL1 is involved in embryonic cell division patterning, root elongation, and pollen tube growth in tobacco. AB - The function of the ARF-GEF family has drawn great attention recently, especially GNOM and GNL1, owing to their important role in plant development. A homolog of GBF was identified in Nicotiana tabacum, named NtGNL1, which is ubiquitously expressed throughout the tobacco life cycle. In NtGNL1 RNAi plants, irregular orientation of cell division and asynchronous cell development during early embryogenesis disrupted the symmetry of the developing embryo. In addition, root growth in transgenic lines was significantly slower than that in wild-type plants, although the structure of the root tip was largely intact. Pollen germination and pollen tube growth were also inhibited in the transgenic lines, and the tip of the pollen tube presented various aberrant morphologies in one of the transgenic lines. The phenotypes of different NtGNL1 RNAi transgenic lines suggest that the NtGNL1 is likely to be involved not only in embryogenesis and postembryonic development, but also in sexual reproduction; thus, NtGNL1 may play multiple and critical roles in plant development. PMID- 18399933 TI - Leaf trichome responses to herbivory in willows: induction, relaxation and costs. AB - To circumvent the inherent problem of discriminating between the cost of losing photosynthetic tissue and the cost of producing an inducible defence, the growth response of herbivore-damaged plants was compared with plants damaged mechanically to the same extent but without eliciting the defence. Two experiments were conducted, studying the response of willows (Salix cinerea) to damage by adult leaf beetles (Phratora vulgatissima). In the first experiment, willows produced new leaves with an enhanced leaf trichome density 10-20 d after damage, coinciding in time with the feeding of beetle offspring. The response was relaxed in foliage produced 30-40 d after damage. In the second experiment, which also included mechanical damage, willows exposed to beetle feeding showed an increase in leaf trichome density of the same magnitude (> 70%) as in the first experiment. The cost of producing the defence was a 20% reduction in shoot length growth and biomass production. Willows exposed to mechanical damage had an 8% reduction in shoot length growth compared with control plants, that is, a cost of leaf area removal. The results are the first quantitative estimates of the cost of a plant defence induced by natural and low amounts (3.3%) of herbivory. PMID- 18399934 TI - Transcriptional profiling of aluminum toxicity and tolerance responses in maize roots. AB - Aluminum (Al) toxicity is a major factor limiting crop yields on acid soils. There is considerable genotypic variation for Al tolerance in most common plant species. In maize (Zea mays), Al tolerance is a complex phenomenon involving multiple genes and physiological mechanisms yet uncharacterized. To begin elucidating the molecular basis of maize Al toxicity and tolerance, a detailed temporal analysis of root gene expression under Al stress was performed using microarrays with Al-tolerant and Al-sensitive genotypes. Al altered the expression of significantly more genes in the Al-sensitive genotype, presumably as a result of more severe Al toxicity. Nevertheless, several Al-regulated genes exhibited higher expression in the Al-tolerant genotype. Cell wall-related genes, as well as low phosphate-responsive genes, were found to be regulated by Al. In addition, the expression patterns of genes related to Al-activated citrate release indicated that in maize this mechanism is probably regulated by the expression level and/or function of the citrate transporter. This study is the first comprehensive survey of global transcriptional regulation under Al stress. The results described here will help to further our understanding of how mechanisms of Al toxicity and tolerance in maize are regulated at the transcriptional level. PMID- 18399935 TI - The long strange trip of Borrelia burgdorferi outer-surface protein C. AB - Borrelia burgdorferi must adapt physiologically to two markedly different host milieus and efficiently transit between its mammalian host and arthropod vector during tick feeding. Differential production of lipoproteins is essential for spirochaetes to survive, multiply and migrate within both hosts. Outer-surface protein C (OspC), which is induced during the blood meal, is critical for transmission of Lyme disease spirochaetes by nymphal ticks. Its biological function is poorly understood, however, despite the fact that its crystal structure has been solved. Evidence has accumulated that OspC blocks clearance of spirochaetes following inoculation in skin, and it is thought to do so by facilitating evasion of innate immunity. The study by Liang and co-workers in this edition of Molecular Microbiology extends this work by showing that OspC prevents early elimination and promotes dissemination. Surprisingly, they also show that unrelated borrelial outer-surface lipoproteins can replace these functions in an ospC mutant. They propose that an abundance of lipoprotein(s) is needed to stabilize the borrelial outer membrane against innate defences. This provocative work clearly runs counter to prevailing orthodoxies of bacterial pathogenesis. It also points the way towards future studies that will clarify the 'partially specific' roles of this enigmatic molecule in Lyme disease pathogenesis. PMID- 18399936 TI - NoxA activation by the small GTPase RacA is required to maintain a mutualistic symbiotic association between Epichloe festucae and perennial ryegrass. AB - Small GTPases of the Rac group play a key regulatory role in NADPH oxidase catalysed production of reactive oxygen species (ROS) in mammals and plants, but very little evidence is available for a corresponding role in fungi. We recently showed that ROS produced by a specific fungal NADPH oxidase isoform, NoxA, are crucial in regulating hyphal morphogenesis and growth in the mutualistic symbiotic interaction between Epichloe festucae and perennial ryegrass. We demonstrate here that E. festucae RacA is required for NoxA activation and regulated production of ROS to maintain a symbiotic interaction. Deletion of racA resulted in decreased ROS production, reduction of radial growth and hyper branching of the hyphae in culture. In contrast, in planta the racA mutant showed extensive colonization of the host plant, resulting in stunting and precocious senescence of the host plants. Strains expressing a dominant active (DA) allele of RacA had increased ROS production, increased aerial hyphae and reduced radial growth. These results demonstrate that RacA plays a crucial role in regulating ROS production by NoxA, in order to control hyphal morphogenesis and growth of the endophyte in planta. PMID- 18399937 TI - Structure of the exosporium and sublayers of spores of the Bacillus cereus family revealed by electron crystallography. AB - We report on the first step in mapping out the spatial location of structural proteins within the exosporium, namely a description of its three-dimensional architecture. Using electron microscopy and image analysis, we have characterized crystalline fragments from the exosporium of Bacillus cereus, B. thuringiensis and B. anthracis strains and identified up to three distinct crystal types. Type I and type II crystals were examined in three dimensions and shown to form arrays of interlinked crown-like structures each enclosing a cavity approximately 26-34 A deep with threefold symmetry. The arrays appear to be permeated by tunnels allowing access from one surface to the other, possibly indicating that the exosporium forms a semi-permeable barrier. The pore size of approximately 23-34 A would allow passage of the endospore germinants, alanine or inosine but not degradative enzymes or antibodies. Thus the structures appear compatible with a protective role for the exosporium. Furthermore the outermost crystalline layer must act as a scaffold for binding the BclA protein that contributes to the 'hairy nap' layer. The array of crowns may also act as a matrix for the binding or adsorption of other proteins that have been identified in the exosporium such as GroEL, immune inhibitor A and arginase. PMID- 18399938 TI - Type IV pili: e pluribus unum? AB - The widespread role of pili as colonization factors in pathogens has long been recognized in Gram-negative bacteria and more recently in Gram-positive bacteria, making the study of these hair-like filaments a perennial hot topic for research. No other pili are found in as many or as diverse bacteria as type IV pili. This is likely a consequence of their ancient origin and unique ability to promote multiple and strikingly different phenotypes such as attachment to surfaces, aggregation, uptake of DNA during transformation, motility, etc. Two decades of investigations in several model species have shed some light on the structure of these filaments and the molecular basis of some of the properties they confer. Moreover, recent discoveries have led to a better knowledge of the genetic basis and molecular mechanisms of type IV pili biogenesis. This brings us a few steps closer to understanding how these filaments are produced, but leaves us wondering whether (as in the famous motto that inspired the title) out of the many models studied will emerge one unifying mechanism. PMID- 18399939 TI - Polarized growth in fungi--interplay between the cytoskeleton, positional markers and membrane domains. AB - One kind of the most extremely polarized cells in nature are the indefinitely growing hyphae of filamentous fungi. A continuous flow of secretion vesicles from the hyphal cell body to the growing hyphal tip is essential for cell wall and membrane extension. Because microtubules (MT) and actin, together with their corresponding motor proteins, are involved in the process, the arrangement of the cytoskeleton is a crucial step to establish and maintain polarity. In Saccharomyces cerevisiae and Schizosaccharomyces pombe, actin-mediated vesicle transportation is sufficient for polar cell extension, but in S. pombe, MTs are in addition required for the establishment of polarity. The MT cytoskeleton delivers the so-called cell-end marker proteins to the cell pole, which in turn polarize the actin cytoskeleton. Latest results suggest that this scenario may principally be conserved from S. pombe to filamentous fungi. In addition, in filamentous fungi, MTs could provide the tracks for long-distance vesicle movement. In this review, we will compare the interaction of the MT and the actin cytoskeleton and their relation to the cortex between yeasts and filamentous fungi. In addition, we will discuss the role of sterol-rich membrane domains in combination with cell-end marker proteins for polarity establishment. PMID- 18399940 TI - Systematic deletion of Salmonella small RNA genes identifies CyaR, a conserved CRP-dependent riboregulator of OmpX synthesis. AB - Post-transcriptional repression of porin synthesis has emerged as a major function of Hfq-dependent, small non-coding RNAs (sRNAs). Many enterobacteria express OmpX-like porins, a family of outer membrane proteins whose physiological roles and structural properties have been studied intensively. While regulatory sRNAs have been identified for most major and many minor porins of Salmonella and Escherichia coli, a post-transcriptional regulator of OmpX levels has never been found. Here, we have taken a 'reverse target search' approach by systematic inactivation of Salmonella sRNA genes, and screening 35 sRNA deletion strains for effects on OmpX synthesis. We have identified the Hfq-dependent CyaR (formerly RyeE) sRNA as an ompX repressor. Global transcriptomic profiling following induction of CyaR expression suggests that ompX mRNA is the primary target of this sRNA under standard growth conditions. The results of phylogenetic and mutational analyses suggest that a conserved RNA hairpin of CyaR, featuring a C rich apical loop, acts to sequester the Shine-Dalgarno sequence of ompX mRNA and to inhibit translational initiation. We have also discovered that cyaR expression is tightly controlled by the cyclic AMP receptor protein, CRP. This represents a new link between porin repression and nutrient availability that is likely to be widely conserved among enterobacteria. PMID- 18399942 TI - Effect of boiling water carcass immersion on aerobic bacteria counts of poultry skin and processed ground poultry meat. AB - This study was conducted to determine the relationship between bacteria destruction on poultry carcass skin and bacteria in raw ground poultry meat from the same carcasses. Immersion time in boiling water of broiler chicken whole carcasses required for maximum reduction of naturally occurring aerobic bacterial count on skin was measured. Treatments for chicken carcasses consisted of immersion in boiling water (approximately 95 degrees C) for 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4 min. Four skin samples taken following treatment and three taken from subsequently ground carcass meat were analyzed for total aerobic plate counts (APC). Analysis of the data indicated a linear increase in bacterial destruction on skin with increased boiling water immersion time from 0 to 4 min. Reduction of skin bacteria to less than 1 log10 occurred at 3 min carcass immersion or longer. The analysis also indicated that treatment with boiling water and removal of skin was effective in reducing bacterial counts in ground meat to similar levels at all treatment times from 0.5 to 4.0 min. Findings from this study indicated that a boiling water immersion intervention and removal of skin could reduce subsequent bacteria contamination of ground meat. This intervention could minimize the risk of pathogen-contaminated primary processed poultry carcasses used in further processing. PMID- 18399941 TI - Antibody response to an anti-rabies vaccine in a dog population under field conditions in Bolivia. AB - Rabies remains an important public health issue in Bolivia, South America. Public concern and fears are most focussed on dogs as the source of rabies. The objective of the present study was to assess immunity of an inactivated suckling mouse brain vaccine against canine rabies used for the official vaccination campaigns under field conditions in an endemic area of rabies in Bolivia. A total of 236 vaccinated and 44 unvaccinated dogs in Santa Cruz de la Sierra, selected using stratified random sampling, were investigated in order to obtain owned dog characteristics and antibody titres against rabies in April 2007. The proportion of vaccinated dogs with an antibody titre exceeded the protection threshold value of 0.5 EU/ml was 58% [95% confidence intervals (CI): 52-65], indicating that vaccination is likely to elicit an antibody response (odds ratio 6.3, 95% CI: 1.2 11.5). The range of geometric mean of antibody titre for vaccinated dogs (0.89 EU/ml; 95% CI: 0.75-1.04) was considered to meet the minimal acceptable level indicating an adequate immune response to the vaccine. However, the titre level was not satisfactory in comparison with the results from other field investigations with inactivated tissue culture vaccines. It is recommended for public health authorities to (1) consider modernizing their vaccine manufacturing method because the level of immunity induced by the current vaccine is comparably low, (2) conduct frequent vaccination campaigns to maintain high levels of vaccination coverage, and (3) actively manage the domestic dog population in the study area, which is largely responsible for rabies maintenance. PMID- 18399943 TI - All creatures great and minute: a public policy primer for companion animal zoonoses. AB - Approximately 63% of US households have at least one pet, a large percentage of which are considered family members. Pet owners can derive substantial physical and psychological benefits from interaction with companion animals. However, pet ownership is not without risks; zoonotic diseases are increasingly drawing the attention of healthcare professionals, policy makers and the general public. While zoonoses of 'traditional' pets are widely recognized and their prevention and treatment factors are generally known, the growing popularity of 'non traditional' pets has the potential to facilitate human exposure to novel zoonoses. However, the greatest risk of zoonoses probably arises from animals taken directly from the wild to serve as pets. Non-governmental organizations, state veterinary associations and others have been calling for increased regulation of animal imports, some proposing that all 'exotics' be banned from the pet trade. Because zoonotic diseases of companion animals are influenced by interacting factors of ecological, technical, socio-economic, and political origin, efforts to minimize their impact need be multi-dimensional, simultaneously addressing both the ecological and socio-political drivers of disease emergence and transmission. This study is intended to serve as a primer for animal care professionals seeking to engage with policy makers and the pet industry on the prevention of companion animal zoonoses. We provide background on the human-animal bond, risks of zoonoses associated with groups of companion animals, and the public policy context, as well as identify the factors needed to build a comprehensive approach to companion animal zoonoses risk management. Also included are examples of innovative, non-regulatory initiatives designed to limit the spread and impact of companion animal zoonoses, including a reptile salmonella poster, the National Reptile Improvement Plan, Habitattitude campaign, Pet Zoonoses Committee, and a wildlife disease surveillance initiative known as Project TripWire. PMID- 18399944 TI - Frequencies of PRNP gene polymorphisms in Vietnamese dairy cattle for potential association with BSE. AB - Summary Since 2004, significant associations between bovine spongiform encephalopathy (BSE) susceptibility in cattle and frequencies of insertion/deletion (ins/del; indel) polymorphisms within the bovine prion protein gene (PRNP) have been reported. In this study, we investigated the frequencies of indel polymorphisms within two variable sites, a 23-bp indel polymorphism in the promoter region (23indel) and a 12-bp indel polymorphism in intron 1 region (12indel), in the PRNP in 206 Vietnamese dairy cattle and seven Japanese BSE affected cattle. In Vietnamese dairy cattle, the frequency distributions of del allele and del/del genotypic polymorphisms in the 23indel site, which are thought to be associated with BSE susceptibility, were significantly higher, whereas the frequencies of del allelic and del/del genotypic polymorphisms in the 12indel site, which have been reported to confer BSE susceptibility, were significantly lower. We have provided evidence that Vietnamese dairy cattle have a unique genetic background in the PRNP gene in comparison with cattle or sires previously reported in other countries. PMID- 18399945 TI - Juvenile dogs as potential sentinels for West Nile virus surveillance. AB - We conducted a study to determine whether juvenile stray dogs could be sentinels for West Nile virus (WNV) surveillance. Seroconversion was detected 6 weeks before the first reported human case. Our findings provide evidence that dogs could be useful sentinels for monitoring areas for evidence of WNV during transmission seasons. PMID- 18399946 TI - Different testosterone levels are associated with ejaculatory dysfunction. AB - INTRODUCTION: The role of testosterone (T) in pathogenesis of ejaculatory symptoms has not been completely clarified. AIM: To evaluate the possible contribution of T and hypogonadism in the control of the ejaculatory reflex, comparing subjects with premature ejaculation (PE) or delayed ejaculation (DE) to those without ejaculatory dysfunction. METHODS: A consecutive series of 2,437 (mean age 51.9 +/- 13.0 years) male patients with sexual dysfunction was studied. MAIN OUTCOME MEASURE: Several hormonal and biochemical parameters were studied, along with the structured interview on erectile dysfunction (SIEDY) structured interview. Hypogonadism were defined when total testosterone (TT) was lower than 10.4 nmol/L. RESULTS: Among the patients studied, 714 (25.9%) and 121 (4.4%) reported PE and DE, respectively. In the youngest age band (25-40 years), subjects with PE reported higher TT and free testosterone (FT) levels when compared to the other groups (subjects with DE or those without PE and DE; P < 0.05 for both). Conversely, in the oldest age band (55-70 years), lower TT and FT levels were observed in DE subjects. Accordingly, patients with PE showed the lowest (12%) and subjects with DE the highest (26%) prevalence of hypogonadism. These differences were confirmed even after adjustment for confounders such as age and libido (HR = 0.75 [0.57-0.99] and 1.83 [1.14-3.94] for PE and DE, respectively; both P < 0.05). CONCLUSIONS: Our data seem to suggest that T plays a facilitatory role in the control of ejaculatory reflex. Both central and peripheral mechanisms have been advocated to explain this association. Clinical studies are currently in progress to further establish the role of T in the ejaculatory dysfunction, attempting to revert DE by androgen administration. PMID- 18399947 TI - Inhibition of Seminal emission is the main cause of anejaculation induced by a new highly selective alpha1A-blocker in normal volunteers. AB - INTRODUCTION: Recent studies have highlighted the influence of alpha1 adrenoceptor antagonists on ejaculatory function. AIM: We evaluated the effect of a new, highly selective alpha1A-blocker, silodosin, on ejaculatory function of normal volunteers. METHODS: The study included 15 healthy male urologists who voluntarily participated in the study. They took 4 mg of silodosin or a placebo twice daily for 3 days in a randomized, double-blind crossover design. MAIN OUTCOME MEASURES: We investigated the ejaculatory volume, sperm count in urine after ejaculation, and fructose concentration in seminal plasma before and after administration of the agents. RESULTS: All volunteers on silodosin had a complete lack of ejaculation. Three days after completion of silodosin, the mean ejaculatory volume recovered to the baseline level. There was no sperm in urine after ejaculation under silodosin administration in any volunteer. CONCLUSIONS: All volunteers on silodosin had anejaculation and did not show post-ejaculate sperm in their urine. The mechanism of ejaculatory dysfunction caused by silodosin is a loss of seminal emission. PMID- 18399948 TI - Does educational status affect a patient's behavior toward erectile dysfunction? AB - INTRODUCTION: Educational status has been investigated rarely as a potential factor affecting the behavior of patients with new onset erectile dysfunction (ED) toward seeking first medical help and subsequent compliance with prescribed phosphodiesterase type 5 inhibitor (PDE5) therapy. AIM: To test whether the educational status of patients with new onset ED and naive to PDE5 therapy may have a significant impact on the delay before seeking first medical help (DSH) and compliance with the suggested PDE5. MAIN OUTCOME MEASURES: Assessing DSH and compliance with PDE5 in new onset ED patients according to their educational status by means of detailed logistic regression analyses. METHODS: Data from 302 consecutive patients with new onset ED and naive to PDE5s were comprehensively analyzed. Patients were segregated according to their educational status into low (elementary and/or secondary school education) and high (high school and/or university degrees) educational levels. Complete data were available for 231 assessable patients. Univariate (UVA) and multivariate (MVA) logistic regression analyses addressed the association between educational status and DSH after adjusting for age, relationship status, and Sexual Health Inventory for Men score. Likewise, UVA and MVA were performed to test the association between educational status and patient compliance with PDE5 at the 9-month median follow up. RESULTS: Median DSH was 24 months (range 1-350; mean 38.1 +/- 42.8). The lower the educational status, the shorter the DSH (P = 0.03). In contrast, a significantly (P < 0.0001) greater proportion of patients with a higher educational status showed compliance with the suggested PDE5 at the 9-month follow-up. Overall, educational status was not an independent predictor of either DSH or patient compliance with PDE5 therapy. CONCLUSIONS: After adjusting for other variables, our findings suggest that in new onset ED patients, educational status does not independently affect the DSH and patient compliance with PDE5 therapy. PMID- 18399949 TI - Personality in Germany and Minnesota: an IRT-based comparison of MPQ self reports. AB - We compared personality across cultures to explore the extent of cultural differences. This study used samples matched on gender and age from Germany and Minnesota to compare traditional scale scores and IRT-based parameters for Tellegen's (1982) Multidimensional Personality Questionnaire scales. Because the samples consisted of twins, we were able to replicate the findings from the subsamples consisting of one member of each pair with the subsamples consisting of their co-twins. When the full scales were considered, Germans were higher in Aggression and Absorption and Minnesotans were higher in Well-being, Control, and Traditionalism, but, except for those in Traditionalism, different item difficulty parameters explained most of these differences. IRT highlighted less than optimal scale properties as well as differentially functioning items. We discuss the theoretical implications of these findings. PMID- 18399950 TI - Going forward by drawing from the past: personal strivings, personally meaningful memories, and personality traits. AB - The present research explored the idea that what people strive for in their daily lives is meaningfully linked to their autobiographical memories and stable personality characteristics. Study 1 showed that (a) personal strivings (both self-reported and content coded) are related to the emotional and motivational content of self-defining memories, (b) personal strivings are related to self esteem and narcissism, and (c) the relation between personal strivings and personality is partially mediated by memory content. Study 2 extended these findings to a longitudinal context and showed that personal strivings and memory content reciprocally influence each other over time. Together, the findings suggest that when people consider how to move forward in their lives, they draw on their past experiences to establish goals for the future and that the way this process plays out is shaped by people's stable personality characteristics. PMID- 18399951 TI - Do dispositional pessimism and optimism predict ambulatory blood pressure during school days and nights in adolescents? AB - We tested the hypotheses that (1) high pessimism and low optimism (LOT-R overall and subscale scores) would predict high ambulatory blood pressure (ABP) level and 24-hour load (percentage of ABP values exceeding the pediatric 95th percentile) among healthy Black and White adolescents (n=201; 14-16 yrs) across 2 consecutive school days and (2) that the relationships for the pessimism and optimism subscales would show nonlinear effects. The hypotheses were confirmed for pessimism but not for optimism. The results suggest that high pessimism may have different effects than low optimism on ABP and that even moderate levels of pessimism may effect blood pressure regulation. These results suggest that optimism and pessimism are not the opposite poles on a single continuum but ought to be treated as separate constructs. PMID- 18399952 TI - Parents' stories of grandparenting concerns in the three-generational family: generativity, optimism, and forgiveness. AB - Adults' level of Eriksonian generativity in midlife has been shown to predict variations in parenting, but there has been less research on its relation to inter generational processes in the three-generational family. As part of a larger study, a sample of 35 Canadian mothers and fathers described a particular, salient child-rearing problem with grandparents when their first-born children were 8 years old. Descriptions were rated for severity of the problem, anger/irritation, optimism about solution, and forgiveness of the grandparent's behavior. Generativity data were collected by a standard questionnaire (the Loyola Generativity Scale of McAdams; McAdams & de St. Aubin, 1992). Results showed few gender differences, though mothers tended to be angrier than fathers with the grandparents. More mature parents were more forgiving than younger parents and saw problems as less serious, as predicted. Finally, parent level of generativity predicted maternal and paternal forgiveness of grandparent behaviors, as well as paternal, but not maternal, optimism about problem outcomes. Parental generativity may thus serve to encourage greater forgiveness and optimism among the generations of the family. PMID- 18399953 TI - Regulating positive and negative emotions in daily life. AB - The present study examined how people regulate their emotions in daily life and how such regulation is related to their daily affective experience and psychological adjustment. Each day for an average of 3 weeks, participants described how they had regulated their emotions in terms of the reappraisal and suppression (inhibiting the expression) of positive and negative emotions, and they described their emotional experience, self-esteem, and psychological adjustment in terms of Beck's triadic model of depression. Reappraisal was used more often than suppression, and suppressing positive emotions was used less than the other three strategies. In general, regulation through reappraisal was found to be beneficial, whereas regulation by suppression was not. Reappraisal of positive emotions was associated with increases in positive affect, self-esteem, and psychological adjustment, whereas suppressing positive emotions was associated with decreased positive emotion, self-esteem, and psychological adjustment, and increased negative emotions. Moreover, relationships between reappraisal and psychological adjustment and self-esteem were mediated by experienced positive affect, whereas relationships between suppression of positive emotions and self-esteem adjustment were mediated by negative affect. PMID- 18399954 TI - Asymmetry in judgments of personality: others are less differentiated than the self. AB - Previous evidence suggests that judges rely more heavily on implicit personality theories when they rate relatively unfamiliar others. One further implication of this evidence is that correlations among traits should be stronger in other ratings than in self-ratings, particularly when (a) judges lack trait relevant information and/or (b) motivational accuracy is low. We tested these predictions by comparing self- versus other ratings on the Big Five in two studies. Study 1 used previously published data to demonstrate clear self/peer differences in the average relations between Big Five dimensions (excluding Extraversion). Study 2 was based on self- versus other ratings in 12 samples. Overall, the intercorrelations among Big Five traits (excluding Extraversion) tended to be significantly stronger in peer ratings than in self-ratings. The most consistent effect involved the relation between Neuroticism and Agreeableness (overall r= .43 and -.29 in the peer ratings and self-ratings, respectively). In addition, as expected, the degree of relation among traits varied depending upon the type of target being rated (i.e., spouse vs. dating partner vs. friend vs. stranger). Implications of these findings are discussed. PMID- 18399955 TI - Perceived parental conditional regard and autonomy support as predictors of young adults' self- versus other-oriented prosocial tendencies. AB - The current research examined the relations of parental conditional regard and autonomy-supportive parenting with levels of internalization and self- versus other-oriented helping tendencies. As predicted from self-determination theory, college students' perceptions of parental conditional regard correlated positively with introjection internalization and self-oriented helping tendencies. Autonomy-supportive parenting predicted fuller internalization and other-oriented helping. Results suggest that the relations between parental practices and prosocial orientations are mediated by the level of internalization that was predicted by each parental practice. PMID- 18399956 TI - Comparing clinical and social-personality conceptualizations of narcissism. AB - There is a lack of consensus surrounding the conceptualization of narcissism. The present study compared two measures of narcissism-one used in clinical settings (Personality Diagnostic Questionnaire, PDQ-4+; Hyler, 1994) and one used in social-personality research (Narcissistic Personality Inventory, NPI; Raskin & Terry, 1988)-across two samples. Sample 1 (N=271) was composed of undergraduates, whereas Sample 2 (N=211) was composed of parents of the Sample 1 participants. The scales were significantly interrelated but manifested divergent relations with general personality traits, personality disorders (including expert prototypal ratings of narcissism), recollections of parenting received, and psychological distress and self-esteem. PDQ-4 narcissism captured an emotionally unstable, negative-affect-laden, and introverted variant of narcissism; NPI narcissism captured an emotionally resilient, extraverted form. The clinical and social-personality conceptualizations of narcissism primarily share a tendency to use an antagonistic interpersonal style. Implications for the DSM-V are discussed. PMID- 18399957 TI - Goal striving within agentic and communal roles: separate but functionally similar pathways to enhanced well-being. AB - Do agency and communion strivings provide functionally similar but predictively independent pathways to enhanced well-being? We tested this idea via a year-long study of 493 diverse community adults. Our process model, based on self determination and motive disposition theories, fit the data well. First, the need for achievement predicted initial autonomous motivation for agentic (work and school) role-goals and the need for intimacy predicted felt autonomy for communal (relationship and parenting) goals. For both agentic and communal goals, autonomous motivation predicted corresponding initial expectancies that predicted later goal attainment. Finally, each type of attainment predicted improved adjustment or role-satisfaction over the year. Besides being similar across agency and communion, the model was also similar across race and gender, except that the beneficial effects of communal goal attainment were stronger for high need for intimacy women and Blacks. Implications for agency/communion theories, motivation theories, and theories of well-being are discussed. PMID- 18399958 TI - Explaining the extraversion/positive affect relation: sociability cannot account for extraverts' greater happiness. AB - The association between Extraversion and positive affect is one of the most robust findings in the study of personality and emotion. Temperament models posit that the association is direct; instrumental models posit that the association is mediated by additional processes. Two experience sampling studies were conducted to test instrumental mechanisms that might underlie the effect. According to a mediation model, extraverts' greater social activity can account for their increased positive affect when compared to introverts. According to a person-by situation interaction model, extraverts react more positively to social situations than do introverts, and this interaction can account for the association. Only weak support for the instrumental models was found; consistent with temperament models, a moderate direct association remained even after controlling for these effects. PMID- 18399959 TI - An immunohistochemical study of the extracellular matrix of entheses associated with the human pisiform bone. AB - The immunohistochemical labelling patterns of the extracellular matrix at the insertion of the flexor carpi ulnaris tendon and the entheses at both ends of the pisometacarpal and pisohamate ligaments were compared in order to relate the molecular composition of the attachment sites to their mechanical environment. Tissue was obtained from elderly dissecting room cadavers and labelled with a panel of monoclonal antibodies directed against collagens, glycosaminoglycans, proteoglycans and matrix proteins. All entheses were fibrocartilaginous and labelled positively for molecules typically associated with articular cartilage (type II collagen, chondroitin 6 sulphate, aggrecan and link protein). Labelling for type II collagen was most conspicuous at the attachment of the flexor carpi ulnaris tendon. In the ligaments, type II collagen labelling was always greater at the pisiform end. Matrilin 1 was universally present at all five entheses examined and fibromodulin labelling was most intense around the tidemark. Fibromodulin may thus be involved in anchorage and/or the control of mineralization at the hard-soft tissue interface of entheses. The greater prominence of fibrocartilage at the pisiform enthesis of the flexor carpi ulnaris tendon than at any ligament attachment may relate to the marked change in the tendon insertional angle that occurs with wrist movements. We also suggest that the more fibrocartilaginous character of the proximal compared with the distal ends of the ligaments relates to the fact that the pisiform is anchored in position and is thus at the centre of rotation of any movement of ligaments attached to it. PMID- 18399960 TI - Comparison of neuroprotective and neurorestorative capabilities of rasagiline and selegiline against lactacystin-induced nigrostriatal dopaminergic degeneration. AB - Nigrostriatal neurodegeneration in Parkinson's disease (PD) has been postulated to be caused by various pathological conditions, such as mitochondrial defects, oxidative stress, and ubiquitin-proteasome system (UPS) dysfunction. Pharmacological strategies designed to interfere with these pathological pathways may effectively counteract the degeneration. Rasagiline and selegiline are selective and irreversible monoamine oxidase-B inhibitors that possess significant protective properties on dopamine neurons in various pre-clinical models of PD. In the present study, the neuroprotective and neurorestorative effects of rasagiline and selegiline were compared in an animal model of PD produced by inhibition of the UPS. C57BL/6 male mice were microinjected bilaterally with UPS inhibitor lactacystin (1.25 mug/side), into the medial forebrain bundle. Administration of rasagiline (0.2 mg/kg, i.p. once per day) or selegiline (1 mg/kg, i.p. once per day), started 7 days before or after (up to 28 days) after lactacystin microinjection. We found that both rasagiline and selegiline exerted a significant neuroprotective effect against lactacystin induced neurodegeneration; but only rasagiline managed to restore the nigrostriatal degeneration. Furthermore, rasagiline showed a modest protection against lactacystin-induced inhibition of proteasomal activity. Our study indicates that compared with selegiline, rasagiline is more potent in protecting neurodegeneration induced by UPS impairment and may, therefore, exert disease modifying effects in PD. PMID- 18399961 TI - Neuromelanin selectively induces apoptosis in dopaminergic SH-SY5Y cells by deglutathionylation in mitochondria: involvement of the protein and melanin component. AB - Parkinson's disease (PD) is characterized by selective depletion of nigral dopamine (DA) neurons containing neuromelanin (NM), suggesting the involvement of NM in the pathogenesis. This study reports induction of apoptosis by NM in SH SY5Y cells, whereas protease-K-treated NM, synthesized DA- and cysteinyl dopamine melanin showed much less cytotoxicity. Cell death was mediated by mitochondria mediated apoptotic pathway, namely collapse of mitochondrial membrane potential, release of cytochrome c, and activation of caspase 3, but Bcl-2 over-expression did not suppress apoptosis. NM increased sulfhydryl content in mitochondria, and a major part of it was identified as GSH, whereas dopamine melanin significantly reduced sulfhydryl levels. Western blot analysis for protein-bound GSH demonstrated that only NM reduced S-glutathionylated proteins in mitochondria and dissociated macromolecular structure of complex I. Reactive oxygen and nitrogen species were required for the deglutathionylation by NM, which antioxidants reduced significantly with prevention of apoptosis. These results suggest that NM may be related to cell death of DA neurons in PD and aging through regulation of mitochondrial redox state and S-glutathionylation, for which NM-associated protein is absolutely required. The novel function of NM is discussed in relation to the pathogenesis of PD. PMID- 18399963 TI - Right ventricular septal lead implantation: new site, new risks? PMID- 18399964 TI - Left ventricular pseudoaneurysm following a "POP" phenomenon. PMID- 18399965 TI - Is there a connection? PMID- 18399966 TI - Mechanism of tissue heating during high intensity focused ultrasound pulmonary vein isolation: implications for atrial fibrillation ablation efficacy and phrenic nerve protection. AB - INTRODUCTION: The purpose of this study was to examine tissue temperatures around pulmonary veins (PVs) during high intensity focused ultrasound (HIFU) balloon ablation for atrial fibrillation. The thermodynamics and extent of PV and phrenic nerve (PN) heating during HIFU ablation have not been established. METHODS AND RESULTS: Tissue temperatures were recorded from epicardial thermocouples at right superior (RS) PV orifice and PN in eight dogs undergoing 51 RSPV HIFU ablations (40 seconds, 40 W). Maximum tissue temperatures, reflected by 288 recording profiles, were negatively correlated with distance from balloon surface (r = 0.79, P < 0.001). Average maximum RSPV temperature was 56.8 +/- 16.8 degrees C (distance: 6.6 +/- 4.1 mm), resulting in full-thickness, circumferential PV isolation in 7 of 8 animals. Direct local mechanical heating restricted to the focused ultrasound zone showed temperature rise to > or =50 degrees C within 10 seconds to a maximum temperature of 82.6 +/- 8.9 degrees C (n = 31). Apparent conductive heating showed gradual heating patterns (<50 degrees C within 10 seconds) at greater distance. PN temperature at all recording sites was 45.0 +/- 11.3 degrees C (distance: 9.2 +/- 5.0 mm). Intentional PN injury (10 of 51 [19.6%] ablations), occurred at 63.7 +/- 16.0 degrees C with closest distance of 4.4 +/- 2.5 mm, which was closer than in PNs without injury (7.3 +/- 4.3 mm, P = 0.005). CONCLUSIONS: HIFU ablation is highly effective in generating temperatures needed for full-thickness, circumferential ablation through rapid direct mechanical heating. Gradual heating of surrounding tissue supports the presence of conductive heating. This study also discloses direct HIFU effects as the mechanism of PN injury occurring within 4-7 mm from balloon surface. PMID- 18399967 TI - Effect of cardiac resynchronization therapy in isolated ventricular noncompaction in adults: follow-up of four cases. AB - BACKGROUND: An isolated ventricular noncompaction (IVNC) is an unclassified cardiomyopathy and, despite the increasing awareness of and interest in this disorder, the role of cardiac resynchronization therapy (CRT) remains obscure. OBJECTIVE: The purpose of this study was to clarify the long-term effect of CRT on IVNC in adult patients. METHODS: Four cases of IVNC were included in this study. Before the CRT device was implanted, all four patients (54 +/- 16-year old, 4 males) presented with symptomatic congestive heart failure. Echocardiography revealed their systolic dysfunction and their left ventricular ejection fraction (LVEF) was 21 +/- 8%. There was also mechanical dyssynchrony observed between the LV septum and free wall area. The QRS duration was "narrow" (112 and 120 ms) in two patients. One patient had been resuscitated from ventricular fibrillation (VF) and two had nonsustained ventricular tachycardia (VT). A CRT defibrillator (CRT-D) was implanted in three patients with VT/VF and a CRT pacemaker (CRT-P) in a patient without VT/VF. The LV lead was positioned in a lateral branch of the coronary sinus where a thickened noncompacted wall existed. RESULTS: During the follow-up period (28 +/- 23 months), their congestive heart failure had improved in terms of the cardiothoracic ratio on the chest X-ray, B-type natriuretic peptide level, LV systolic dimension, and LVEF. No episodes of defibrillation shocks were observed. CONCLUSION: CRT may improve the prognosis and quality-of-life in patients with an IVNC with mechanical dyssynchrony. PMID- 18399968 TI - The optimized V-V interval determined by interventricular conduction times versus invasive measurement by LVdP/dtMAX. AB - INTRODUCTION: We compared the calculated optimal V-V interval derived from intracardiac electrograms (IEGM) with the optimized V-V interval determined by invasive measurement of LVdP/dt(MAX). METHODS AND RESULTS: Thirty-two patients with heart failure (six females, ages 68 +/- 7.8 years) had a CRT device implanted. After implantation of the atrial, right and a left ventricular lead, the optimal V-V interval was calculated using the QuickOpt formula (St. Jude Medical, Sylmar, CA, USA) applied to the respective IEGM recordings (V-V(IEGM)), and also determined by invasive measurement of LVdP/dt(MAX) (V-V(dP/dt)). The optimal V-V(IEGM) and V-V(dP/dt) intervals were 52.7 +/- 18 ms and 24.0 +/- 33 ms, respectively (P = 0.017), without correlation between the two. The baseline LVdP/dt(MAX) was 748 +/- 191 mmHg/s. The mean value of LVdP/dt(MAX) at invasive optimization was 947 +/- 198 mmHg/s, and at the calculated optimal V-V(IEGM) interval 920 +/- 191 mmHg/s (P < 0.0001). In spite of this significant difference, there was a good correlation between both methods (R = 0.991, P < 0.0001). However, a similarly good correlation existed between the maximum value of LVdP/dt(MAX) and LVdP/dt(MAX) at a fixed V-V interval of 0 ms (R = 0.993, P < 0.0001), or LVdP/dt(MAX) at a randomly selected V-V interval between 0 and +80 ms (R = 0.991, P < 0.0001). CONCLUSION: Optimizing the V-V interval with the IEGM method does not yield better hemodynamic results than simultaneous BiV pacing. Although a good correlation between LVdP/dt(MAX) determined with V-V(IEGM) and V V(dP/dt) can be constructed, there is no correlation with the optimal settings of V-V interval in the individual patient. PMID- 18399970 TI - LQTS: better with age? PMID- 18399969 TI - Inducibility of atrial and ventricular arrhythmias along the ligament of marshall: role of autonomic factors. AB - BACKGROUND: The mechanism(s) underlying atrial fibrillation (AF) initiation along the ligament of Marshall (LOM) remains controversial. OBJECTIVES: We sought to examine the role of the autonomic nervous system in arrhythmogenesis along the LOM. METHODS: In 31 anesthetized dogs, a left thoracotomy exposed the LOM. During atrial pacing, high-frequency stimulation (HFS: 200 Hz, 0.1 ms pulse width, 40 ms duration, 0.6-12 V) was delivered during atrial refractoriness to different sites of LOM (LOM(CS)= near coronary sinus; LOM(LIPV)= near left inferior pulmonary vein; LOM(LS-LIPV)= between LIPV and left superior pulmonary vein (LSPV); LOM(LSPV)= near LSPV). HFS was repeated after intravenous administration of esmolol (1 mg/kg; n = 9) or atropine (2 mg; n = 12). Norepinephrine (10(-7) M, 0.4 cc) was injected into LOM(LSPV) (n = 5). RESULTS: The median voltages for HFS to induce AF were 3.2 V, 3.2 V, 8.0 V*(,double dagger), and 12 V*(,double dagger) at LOM(CS), LOM(LIPV), LOM(LS-LIPV), and LOM(LSPV), respectively (*P < 0.01, compared with LOM(CS) and double dagger P < 0.01, compared with LOM(LIPV)). Esmolol or atropine markedly increased the threshold for AF induction. Ventricular tachycardias (VT) and accelerated junctional rhythm were induced in 8 of 12 and 6 of 12 dogs after atropine administration, respectively. Sustained VT occurred within minutes in 5 of 5 dogs receiving norepinephrine injection into the LOM(LSPV.) CONCLUSION: HFS induced AF along LOM with a gradient of stimulation thresholds from LOM(CS) (lowest) toward LOM(LSPV) (highest). This response was inhibited by esmolol or atropine. These data suggest an autonomic basis for AF initiation in LOM, and both sympathetic and parasympathetic neural elements play an important role in AF initiation. Hyperactivity of the sympathetic neural elements in LOM may be crucial in the initiation of ventricular tachyarrhythmias. PMID- 18399971 TI - Atrial tachyarrhythmia: what is the ideal site for successful ablation? PMID- 18399972 TI - Quantitative analysis of the duration of slow conduction in the reentrant circuit of ventricular tachycardia after myocardial infarction. AB - BACKGROUND: Few data are available to define the circuits in ventricular tachycardia (VT) after myocardial infarction and the conduction time (CT) through the zone of slow conduction (SCZ). This study assessed the CT of the SCZ and identified different reentrant circuits. METHODS: During VTs, concealed entrainment (CE) was attempted. The SCZ was identified by a difference between postpacing interval (PPI) and VT cycle length (VTcl) < or =30 ms. Since the CT in the normally conducting part of the VT circuit is constant during VT and CE, a CE site within the reentrant circuit with (S-QRS)/PPI > or = 50% was classified as an inner reentry in which the entire circuit was within the scar, and a CE site with (S-QRS)/PPI < 50% as a common reentry in which part of the circuit was within the scar and part out of the scar. RESULTS: CE was achieved in 20 VTs (12 patients). Six VTs (30%) with a (S-QRS)/PPI > or =50% were classified as inner reentry and 14 VTs (70%) with a (S-QRS)/PPI <50% during CE mapping as common reentry. The EG-QRS interval (308 +/- 73 ms vs 109 +/- 59 ms, P < 0.0001) was significantly longer and the incidence of systolic potentials higher (4/6 vs 0/12, P < 0.001) in the inner reentry group. For the 14 VTs with a common reetry, the CT of the SCZ was 348 +/- 73 ms, while the CT in the normal area was 135 +/- 50 ms. CONCLUSION: According to the proposed classification, 30% of VTs after myocardial infarction had an entire reentrant circuit within the scar. In VTs with a common reentrant circuit, the CT of the SCZ is approximately four times longer than the CT in the normal area, accounting for more than 70% of VTcl. PMID- 18399973 TI - When an ICD is not the answer... hypothyroidism-induced cardiomyopathy and torsades de pointes. AB - INTRODUCTION: An increasing number of patients with left ventricular (LV) dysfunction are referred for placement of an implantable cardioverter defibrillator (ICD). CASE REPORT: A 78-year-old female with fatigue, palpitations, and presyncope was referred for consideration of an ICD because of a cardiomyopathy and nonsustained ventricular tachycardia (VT). Her evaluation revealed severe hypothyroidism, marked QT prolongation, and episodes of torsades de pointes. With levothyroxine therapy, her ventricular arrhythmias rapidly abated, with subsequent normalization of LV function and the QT interval. CONCLUSIONS: This report highlights the critical importance of detecting hypothyroidism as an unusual cause for reversible cardiomyopathy and ventricular arrhythmias. PMID- 18399974 TI - Premature ventricular contractions arising from the pulmonary artery: the use of multielectrode array mapping and cryoablation. AB - Premature ventricular contractions with a left bundle branch block-like morphology and inferior axis typically originate from the right ventricular outflow tract; however, they can also originate from the pulmonary artery. We report two patients with frequent premature ventricular contractions who underwent successful cryoablation of ectopics arising from the posterolateral region of the pulmonary artery. The safe and effective use of cryoablation within the pulmonary artery is discussed. We recommend the use of noncontact mapping with multielectrode array catheter in facilitating the technique and providing a more accurate localization of the ectopic focus within the pulmonary artery, compared with conventional pace mapping methods. PMID- 18399975 TI - Isolation, characterization and identification of lactic acid bacteria and yeasts from sour Mifen, a traditional fermented rice noodle from China. AB - AIMS: Considering the effect of natural fermentation on the textural improvement of fermented rice noodles in China and South Asia, and given the lack of reports concerning microbial populations and structure in the fermentation process, this study aims to determine the number of viable micro-organisms and identify the species isolated from the local factories, and to assess their potential use as a starter culture from their enzymatic profiles. METHODS AND RESULTS: Fourteen samples from three local factories were analysed for the presence of micro organisms. A total of 170 lactic acid bacteria (LAB) and 96 yeasts were isolated from the factories. The isolates were phenotypically characterized by using API 50 CHL kits, API 20 Strep kits, API ID 32 C kits and by performing additional biochemical tests. The enzymatic profiles of isolates were assessed by using API ZYM kits. Lactobacillus plantarum and Saccharomyces cerevisiae were identified as predominant species in the fermented supernatants. A majority of the isolates of LAB and yeasts displayed activities of alpha-glucosidase, beta-glucosidase, lipase and trypsin. CONCLUSIONS: The microbial composition and strain characteristics present in the fermentation supernatant demonstrate that a majority of micro-organisms have the ability to digest starch, sugar, protein or lipid. It supports our previous work in which the rice starch was modified and purified by fermentation and thus improves the texture of rice noodles. SIGNIFICANCE AND IMPACT OF THE STUDY: The dominant strains would be important in developing a starter culture. The results can form the basis for the improvement of product quality and consistency. PMID- 18399976 TI - Salmonella must be viable in order to attach to the surface of prepared vegetable tissues. AB - AIMS: The aims of the current study were to explore the site of bacterial attachment to vegetable tissues and to investigate the hypothesis that Salmonella must be living in order to attach to this site(s). METHODS AND RESULTS: Scanning electron micrographs of intact potato cells showed that Salm. serotype Typhimurium attached to cell-wall junctions; suggesting a high-level of site selectivity. Inactivation of Salm. Typhimurium using heat, ethanol, formalin or Kanamycin resulted in cells that could be no longer attached to these sites. Attachment of a Gfp(+) strain of Salm. Typhimurium to cell-wall material (CWM) was examined via flow cytometric analysis. Only live Salm. Typhimurium attached to the CWM. CONCLUSIONS: Salmonella serotype Typhimurium must be metabolically active to ensure attachment to vegetable tissues. Attachment preferentially occurs at the plant cell-wall junction and the cell-wall components found here, including pectate, may provide a receptor site for bacterial attachment. SIGNIFICANCE AND IMPACT OF THE STUDY: Further studies into individual plant cell wall components may yield the specific bacterial receptor site in vegetable tissues. This information could in turn lead to the development of more targeted and effective decontamination protocols that block this site of attachment. PMID- 18399977 TI - Dietary glycated protein modulates the colonic microbiota towards a more detrimental composition in ulcerative colitis patients and non-ulcerative colitis subjects. AB - AIM: To investigate the effect of native, heated and glycated bovine serum albumin (BSA) on the ulcerative colitis (UC) and non-UC colonic microbiota in vitro. METHODS AND RESULTS: Continuous flow culture (CFC) models of the human colonic microbiota inoculated with faeces from UC and non-UC volunteers were maintained on BSA as growth substrate. Changes in bacterial populations and short chain fatty acids were determined. UC and non-UC microbiota differed significantly in microbial populations, with elevated numbers of sulfate-reducing bacteria (SRB) and clostridia in the microbiota from UC patients. Compared with native BSA, glycated BSA modulated the gut microbiota of UC patients in vitro towards a more detrimental community structure with significant increases in putatively harmful bacteria (clostridia, bacteroides and SRB; P < 0.009) and decreases in dominant and putatively beneficial bacterial groups (eubacteria and bifidobacteria; P < 0.0004). The levels of beneficial short-chain fatty acids were significantly decreased by heated or glycated BSA, but were increased significantly by native BSA. CONCLUSION: The UC colonic microbiota maintained in CFC was significantly modified by glycated BSA. SIGNIFICANCE AND IMPACT OF THE STUDY: Results suggest that dietary glycated protein may impact upon the composition and activity of the colonic microbiota, an important environmental variable in UC. PMID- 18399978 TI - UV mutagenesis of Cupriavidus necator for extracellular production of (R)-3 hydroxybutyric acid. AB - AIM: Ultraviolet (UV) mutagenesis was carried out to obtain mutant strains of Cupriavidus necator that could produce (R)-3-hydroxybutyric acid [(R)-3-HB] in the culture supernatant. METHODS AND RESULTS: C. necator (formerly known as Ralstonia eutropha) was subjected to UV radiation to generate mutants that are capable of producing (R)-3-HB in the culture supernatant. Results indicated that UV mutagen disrupted the phbB (phbB knock-out) and thus, promoted production of (R)-3-HB in mutant strains. Inclusion of acetoacetate esters (carbonyl compounds) in the culture broth led to increased production of (R)-3-HB. Thus, acetoacetyl CoA (an intermediate of the PHB synthetic pathway) might have been converted to acetoacetate, which in the presence of (R)-3-HB dehydrogenase and NADPH/NADP(+), resulted in extracellular production of (R)-3-HB. CONCLUSIONS: UV mutagenesis proved to be a satisfactory method in generating interesting mutants for extracellular production of (R)-3-HB. Extracellular production of (R)-3-HB upon addition of acetoacetate esters would suggest a likely (R)-3-HB biosynthetic pathway in C. necator. SIGNIFICANCE AND IMPACT OF THE STUDY: Mutants obtained in this study are very useful for production of (R)-3-HB. For the first time, the production of (R)-3-HB by C. necator via acetoacetate is reported. PMID- 18399979 TI - An evolutionary perspective on Y-chromosomal variation and male infertility. AB - Genetic variation on the Y chromosome is one of the best-documented causes of male infertility, but the genes responsible have still not been identified. This review discusses how an evolutionary perspective may help with interpretation of the data available and suggest novel approaches to identify key genes. Comparison with the chimpanzee Y chromosome indicates that USP9Y is dispensable in apes, but that multiple copies of TSPY1 may have an important role. Comparisons between infertile and control groups in search of genetic susceptibility factors are more complex for the Y chromosome than for the rest of the genome because of population stratification and require unusual levels of confirmation. But the extreme population stratification exhibited by the Y also allows populations particularly suitable for some studies to be identified, such as the partial AZFc deletions common in Northern European populations where further dissection of this complex structural region would be facilitated. PMID- 18399980 TI - Construction of cavernosum smooth muscle using umbilical artery smooth muscle cells seeded on acellular corporal collagen matrices. AB - The objective of this study was to investigate the feasibility of tissue engineering of corpus cavernosal smooth muscle. Acellular corporal collagen matrices (ACCMs) were obtained from the penis of adult rabbits by a cell removal procedure. ACCMs were implanted into the back muscles of allogenic rabbits to investigate the resulting immunological reaction. Human umbilical artery smooth muscle cells (HUASMCs) were isolated from human umbilical arteries through explant techniques and expanded in vitro. Subsequently, third and fifth passage HUASMCs were seeded to ACCMs at a concentration of 30 x 10(6) cells/mL. Then, seeded ACCMs were implanted subcutaneously in athymic mice. The implants were retrieved at 10, 20 and 40 days after implantation. Histochemistry, immunohistochemistry and scanning electron microscopy were performed to analyse the morphological characteristics of the engineered tissues. Additionally, organ bath studies were performed to address the contractility of the engineered tissues. The decellularization process successfully extracted all cellular components while preserving the original collagen fibers. The immunological reaction to ACCMs consisted of only a transient nonspecific inflammatory response. Light and scanning electron microscopy demonstrated that HUASMCs extended onto the three-dimensional ACCMs scaffolds in vitro. Histological analyses of the explants from all time points demonstrated a progressive regeneration of smooth muscle, with structures very similar to native corpus cavernosum smooth muscle. The maximum contraction force induced by phenylephrine and electrical stimulation were 3.64 +/- 0.18 g/100 mg and 2.50 +/- 0.21 g/100 mg, respectively. Our study demonstrates that HUASMCs can be seeded on three dimensional ACCM scaffolds and will develop tissues similar to that of the native corpus cavernosum smooth muscle. PMID- 18399981 TI - PSP-I/PSP-II spermadhesin exert a decapacitation effect on highly extended boar spermatozoa. AB - PSP-I/PSP-II heterodimer is a major protein of boar seminal plasma that is able to preserve, in vitro, the viability, motility and mitochondrial activity of highly-extended boar spermatozoa. However, a relationship between the protective effects of the heterodimer and sperm capacitation is still unclear. The present study investigated the effect of the PSP-I/PSP-II (1.5 mg/mL) on membrane stability, intracellular calcium concentration ([Ca(2+)](I)) and plasma membrane and acrosome integrity of highly extended boar spermatozoa. Boar spermatozoa were diluted to 1 x 10(6) spermatozoa/mL and incubated at 38 degrees C in Phosphate buffered saline (PBS) for 10, 30, 60, 120 and 300 min or in modified Tris buffered medium (mTBM) for 10, 20, 30, 60 and 120 min. After each incubation time, the membrane stability (using Merocyanine-540/Yo-Pro-1), elevation of [Ca(2+)](I) (using Fluo-3-AM/PI) and the sperm plasma membrane and acrosome integrity (using SYBR-14/PI/PE-PNA) were evaluated by flow cytometry. As expected, exposure of the spermatozoa to the PSP-I/PSP-II preserved the plasma membrane and acrosome integrity compared to non-exposed spermatozoa in both media PBS and mTBM (p < .01). The evaluation of membrane stability showed no differences in the percentages of viable sperm with instable plasma membrane in the presence of the PSP-I/PSP-II compared to controls irrespective of the dilution media. The evaluation of the [Ca(2+)](I) levels showed that while spermatozoa incubated in mTBM and exposed to PSP-I/PSP-II had lower [Ca(2+)](I) than controls (39.08% vs. 47.97%, respectively; p < .05), no differences were observed in those samples incubated in PBS. However, a temporal evaluation of the samples showed that a similar proportion of live spermatozoa were able to achieve high levels of [Ca(2+)](I) and membrane instability independent of the presence of PSP-I/PSP-II. In conclusion, PSP-I/PSP-II exert a non-permanent decapacitation effect on highly extended boar spermatozoa that is related with a delay in the increase of [Ca(2+)](I) levels. PMID- 18399982 TI - Predictive value of CASA parameters in IUI with frozen donor sperm. AB - The objective of this study was to determine if characteristics of sperm motion determined by computer-aided semen analysis (CASA) after thawing and preparation on discontinuous gradient could predict pregnancy outcome after intrauterine insemination (IUI) from frozen donor sperm. A retrospective analysis of 100 non selected women undergoing 171 consecutive donor insemination cycles was conducted between January 2006 and April 2007. Semen samples from all donors were analysed after thawing and density gradient preparation. Women who became pregnant and those who did not were comparable in terms of age, ovarian stimulation regimen and indication of IUI with donor semen. Pregnancy rate per cycle was 21.8%, and pregnancy occurred after 2.5 IUI cycles on average. Motility parameters of sperm measured by CASA (VAP, VCL, VSL, LIN, STR, and ALH) and total spermatozoa concentration after preparation on discontinuous gradient showed no difference in both groups. Progressive and total motile spermatozoa concentration, as well as progressive and total motile percentages was significantly higher in pregnancy group. The receiver operating characteristic (ROC) curve analysis showed that total motile percentage >17% and motile concentration >0.9 x 10(6)/mL best predicted pregnancy. In a multivariate analysis, only total motility percentage was able to predict pregnancy. Sperm motility parameters of frozen-thawed prepared donor sperm obtained by CASA do not seem to predict pregnancy in IUI cycles. Total motile and progressive percentages and concentrations remain the best prognostic elements for pregnancy in IUI with donor semen. PMID- 18399983 TI - Capacitation-associated changes in membrane fluidity in asthenozoospermic human spermatozoa. AB - The fertilizing potential of human spermatozoa relies on their ability to capacitate as they travel through the female reproductive tract. During this process, cholesterol is released from the plasma membrane, altering its architecture and dynamics. Using ISolate gradients, we obtained high (L90)- and low (L45)-quality spermatozoa from asthenozoospermic human semen samples. We tested the hypothesis that the lower fertilizing ability of asthenozoospermic L90 cells could be related to a lower ability to increase their membrane fluidity during capacitation. We assessed two sets of fluorescent probes: (i) DPH, TMA-DPH and PA-DPH which senses the hydrophobic core, cytosolic and exofacial leaflets of the bilayer, respectively and (ii) Laurdan, sensitive to the amount of water molecules intercalated between lipid moieties of the membrane (membrane hydration). Before capacitation, membrane fluidity of asthenozoospermic sperm populations was similar to the corresponding fractions of normozoospermic cells when evaluated with DPH, TMA-DPH or PA-DPH. Asthenozoospermic whole samples displayed lower plasma membrane hydration than normozoospermic cells as evidenced with Laurdan. After capacitation, asthenozoospermic L45 and L90 cells failed to increase their membrane fluidity in opposition to normozoospermic cells. Interestingly, membrane hydration significantly correlated with the main sperm motion parameters analysed, being a low membrane hydration associated with poor sperm movement. These results show that low-motility spermatozoa are unable to respond to capacitation with the necessary changes in membrane fluidity. This defect in sperm plasma membrane rheology may be responsible for their poor functional quality and low fertilizing ability. PMID- 18399984 TI - Prophylactic effect of recombinant factor VIIa with congenital factor VII deficiency. PMID- 18399985 TI - Haplotype analysis confirms the association between the HCRTR2 gene and cluster headache. AB - BACKGROUND: Several studies suggested that genetic factors play a role in cluster headache (CH) susceptibility. We found a significant association between the 1246 G>A polymorphism of the hypocretin receptor-2 (HCRTR2) gene and the disease. This association was confirmed in a large study from Germany but was not replicated in a dataset of CH patients from Northern Europe. OBJECTIVE: The purpose of this study was to further evaluate the association between CH and the HCRTR2 gene using new polymorphisms, estimating the frequency of different gene haplotypes, searching for gene mutations, and evaluating the effects of the examined polymorphisms on hypocretin binding sites. METHODS: We genotyped 109 CH patients and 211 healthy controls for 5 new polymorphisms of the HCRTR2 gene and we inferred different gene haplotypes. Complete HCRTR2 sequencing was undertaken for 11 independent CH patients, 5 of whom had a positive family history. The effects of the 1246 G>A polymorphism on the hypocretin binding sites were evaluated using different computer-assisted analyses. RESULTS: Three new polymorphisms of the HCRTR2 gene resulted significantly associated with CH. The GTAAGG haplotype resulted more frequent in cases than in controls (OR: 3.68; 95% CI: 1.85-7.67). No point mutation of the HCRTR2 gene was found. Binding analyses showed that the 1246 G>A polymorphism (substitution of valine at position 308 by isoleucine) has no effect on the hypocretin binding sites but could influence the dimerization process of the receptor. CONCLUSION: Our data confirm previous studies suggesting that the HCRTR2 gene or a linked locus significantly modulates the risk for CH. In addition, we suggest that the V308I substitution of the HCRTR2 may interfere with the dimerization process of the receptor, thereby influencing its functional activity. PMID- 18399986 TI - Candida krusei produces ethanol without production of succinic acid; a potential advantage for ethanol recovery by pervaporation membrane separation. AB - The development of fermentative yeasts secreting no organic acids is highly desirable for ethanol production coupled with membrane separation processes, because the acidic byproduct, succinic acid, significantly inhibits the membrane permeation of ethanol. Of the Pichia and Candida yeasts tested, Candida krusei IA 1 showed the highest ethanol productivity [55 g L(-1) day(-1) from 150 g L(-1) (w/v) of glucose], comparable to the strains of Saccharomyces cerevisiae, and produced much less of the acid (0.6 g L(-1) day(-1)) than the Saccharomyces strains (1.5-1.8 g L(-1) day(-1)) under semi-aerobic conditions. Interestingly, under aerobic conditions, strain IA-1 showed no production of the acid. Stain IA 1 exhibited a good assimilation of the acid, while S. cerevisiae NBRC 0216 showed no assimilation. The activity of succinate dehydrogenase (SDH) in strain IA-1 was 37.5 mU mg(-1), and 7.8-fold higher than that in S. cerevisiae strain NBRC 0216. More significantly, SDH1 was abundantly transcribed in strain IA-1, different from that in strain NBRC 0216, regardless of the culture conditions. From these results, C. krusei IA-1 efficiently takes up succinic acid and metabolizes it in the Krebs cycle, producing an extremely low level of byproducts in the culture medium. Therefore, C. krusei is not only a promising alternative to S. cerevisiae but also a suitable model for metabolic engineering of S. cerevisiae. PMID- 18399987 TI - Carbonyl cyanide m-chlorophenylhydrazone induced calcium signaling and activation of plasma membrane H(+)-ATPase in the yeast Saccharomyces cerevisiae. AB - The plasma membrane H(+)-ATPase from Saccharomyces cerevisiae is an enzyme that plays a very important role in the yeast physiology. The addition of protonophores, such as 2,4-dinitrophenol (DNP) and carbonyl cyanide m chlorophenylhydrazone (CCCP), also triggers a clear in vivo activation of this enzyme. Here, we demonstrate that CCCP-induced activation of the plasma membrane H(+)-ATPase shares some similarities with the sugar-induced activation of the enzyme. Phospholipase C and protein kinase C activities are essential for this activation process while Gpa2p, a G protein involved in the glucose-induced activation of the ATPase, is not required. CCCP also induces a phospholipase C dependent increase in intracellular calcium. Moreover, we show that the availability of extracellular calcium is required for CCCP stimulation of H(+) ATPase, suggesting a possible connection between calcium signaling and activation of ATPase. PMID- 18399989 TI - Mycoplasma gallisepticum as the first analyzed bacterium in which RNA is not polyadenylated. AB - The addition of poly(A)-tails to RNA is a phenomenon common to almost all organisms. In addition to most eukaryotic mRNAs possessing a stable poly(A)-tail, RNA is polyadenylated as part of a degradation mechanism in prokaryotes, organelles, and the eukaryotic nucleus. To date, only very few systems have been described wherein RNA is metabolized without polyadenylation, including several archaea and yeast mitochondria. The minimal genome of the parasitic bacteria, Mycoplasma, does not encode homologs of any known polyadenylating enzyme. Here, we analyze polyadenylation in Mycoplasma gallisepticum. Our results suggest this organism as being the first described bacterium in which RNA is not polyadenylated. PMID- 18399988 TI - The endogenous adrenodoxin reductase-like flavoprotein arh1 supports heterologous cytochrome P450-dependent substrate conversions in Schizosaccharomyces pombe. AB - Mitochondrial cytochromes P450 are essential for biosynthesis of steroid hormones, vitamin D and bile acids. In mammals, the electrons needed for these reactions are provided via adrenodoxin and adrenodoxin reductase (AdR). Recently, Schizosaccharomyces pombe was introduced as a new host for the functional expression of human mitochondrial steroid hydroxylases without the coexpression of their natural redox partners. This fact qualifies S. pombe for the biotechnological production of steroids and for application as inhibitor test organism of heterologously expressed cytochromes P450. In this paper, we present evidence that the S. pombe ferredoxin reductase, arh1, and ferredoxin, etp1fd provide mammalian class I cytochromes P450 with reduction equivalents. The recombinant reductase showed an unusual weak binding of flavin adenine dinucleotide (FAD), which was mastered by modifying the FAD-binding region by site-directed mutagenesis yielding a stable holoprotein. The modified reductase arh1_A18G displayed spectroscopic characteristics similar to AdR and was shown to be capable of accepting electrons with no evident preference for NADH or NADPH, respectively. Arh1_A18G can substitute for AdR by interacting not only with its natural redox partner etp1fd but also with the mammalian homolog adrenodoxin. Cytochrome P450-dependent substrate conversion with all combinations of the mammalian and yeast redox proteins was evaluated in a reconstituted system. PMID- 18399990 TI - Attempts to revive Mycobacterium tuberculosis from 300-year-old human mummies. AB - Environmental persistence of Mycobacterium tuberculosis is subject to speculation. However, the reality that infected postmortem tissues can be a danger to pathologists and embalmers has worrisome implications. A few experimental studies have demonstrated the organism's ability to withstand exposure to embalming fluid and formalin. Recently, a failure was reported in an attempt to resuscitate an original isolate of Robert Koch to determine the lifetime of the tubercle bacillus. The present study also considers a historical approach to determine persistence under favorable environmental conditions. It asks whether acid-fast forms observed in tissues of 300-year-old Hungarian mummies can be resuscitated. Finding organisms before the advent of antibiotics and pasteurization may yield valuable genetic information. Using various media modifications, as well as guinea pig inoculation, an attempt was made to culture these tissues for M. tuberculosis. In addition, a resuscitation-promoting factor, known to increase colony counts in high G+C bacteria, was applied to the cultures. Although an occasional PCR-positive sample was detected, no colonies of M. tuberculosis were obtained. Our results may indicate that the life span of the tubercle bacillus is less than a few hundred years, even though in the short run it can survive harsh chemical treatment. PMID- 18399991 TI - Update on macrolide-lincosamide-streptogramin, ketolide, and oxazolidinone resistance genes. AB - This Minireview summarizes the changes in the field of bacterial resistance to macrolide, lincosamide, streptogramin, ketolide, and oxazolidinone (MLSKO) antibiotics since the nomenclature review in 1999. A total of 66 genes conferring resistance to this group of antibiotics has now been identified and includes 13 new rRNA methylase genes, four ATP-binding transporter genes coding for efflux proteins, and five new inactivating enzymes. During this same time period, 73 new genera carrying known rRNA methylase genes and 87 new genera carrying known efflux and/or inactivating genes have been recognized. The number of bacteria with mutations in the genes for 23S rRNA, L4 and L22 ribosomal proteins, resulting in reduced susceptibility to some members of the group of MLSKO antibiotics has also increased and now includes nine different Gram-positive and 10 different Gram-negative genera. New conjugative transposons carrying different MLSKO genes along with an increased number of antibiotics and/or heavy metal resistance genes have been identified. These mobile elements may play a role in the continued spread of the MLSKO resistance genes into new species, genera, and ecosystems. PMID- 18399992 TI - Photoreactivation of (6-4) photolyase in Dunaliella salina. AB - Dunaliella salina is a unicellular green alga and possesses two types of photolyase: Class II cyclobutane pyrimidine dimers (CPD) photolyase and (6-4) photolyase. The gene of D. salina (6-4) photolyase is the first one found in unicellular organisms. CPD photolyases have been extensively studied but (6-4) photolyases are less understood. Because of the data observed in this study, D. salina (6-4) photolyase is insensitive to high salinity; whether it can tolerate a higher level of salinity than other (6-4) photolyases needs to be studied further. However, evidence is provided that (6-4) photolyases might be highly conserved among different species, not only in the sequence identity but also in the photorepair mechanism. PMID- 18399993 TI - Stable low molecular weight RNA profiling showed variations within Sinorhizobium meliloti and Sinorhizobium medicae nodulating different legumes from the alfalfa cross-inoculation group. AB - Four different low molecular weight (LMW) RNA profiles, designated I-IV, among 179 isolates from Medicago, Melilotus and Trigonella species growing in a field site in Northern Spain were identified. From sequence analysis of the 16S rRNA, atpD and recA genes as well as DNA-DNA hybridization analysis with representatives of each LMW RNA profile it was evident that isolates with LMW RNA profiles I and II belonged to Sinorhizobium meliloti and those displaying profiles III and IV to Sinorhizobium medicae. Therefore, two distinct LMW RNA electrophoretic mobility profiles were found within each of these two species. Collectively, LMW RNA profiles I and II (identified as S. meliloti) were predominant in Melilotus alba, Melilotus officinalis and Medicago sativa. Profiles III and IV (identified as S. medicae) were predominant in Melilotus parviflora, Medicago sphaerocarpa, Medicago lupulina and Trigonella foenum graecum. All the four LMW RNA profiles were identified among isolates from Trigonella monspelliaca nodules. These results revealed a different specificity by the hosts of the alfalfa cross-inoculation group towards the two bacterial species found in this study. PMID- 18399994 TI - Isolation of a thermophilic and halophilic tyrosol-degrading Geobacillus from a Tunisian high-temperature oil field. AB - An aerobic, thermophilic, halotolerant and Gram-positive bacterium, designated strain C5, was isolated from a high-temperature oil field, located in Sfax, Tunisia, after enrichment on tyrosol. Strain C5 grew between 25 and 70 degrees C and optimally at 50 degrees C. It grew in the presence of 0-12% (w/v) NaCl, with optimum growth at 3% (w/v) NaCl. Strain C5 was able to degrade tyrosol aerobically, in the presence of 30 g L(-1) NaCl and under warm conditions (55 degrees C). The degradation of tyrosol proceeded via p-hydroxyphenylacetic and 3,4-dihydroxyphenylacetic acids. The products were confirmed by HPLC and GC-MS analyses. Strain C5 was also found to degrde a wide range of other aromatic compounds, including benzoic, p-hydroxybenzoic, protocatechuic, vanillic, p hydroxyphenylacetic, 3,4-dihydroxyphenylacetic, cinnamic and ferulic acids, phenol and m-cresol. Moreover, strain C5 was grown on diesel and crude oil as sole carbon and energy sources. Strain C5 was also able to utilize several carbohydrates. Phenotypic characteristics and phylogenetic analysis of the 16S rRNA gene sequence of strain C5 revealed that it was related to members of the genus Geobacillus, being most closely related to the type strain of G. pallidus (99% sequence similarity). In addition, we report on growth of the type strain of G. pallidus on different aromatic compounds and hydrocarbons. PMID- 18399995 TI - Nitrogen source influences natural abundance (15)N of Escherichia coli. AB - Escherichia coli cells were forced to mineralize or assimilate nitrogen in vitro by manipulating substrate carbon and nitrogen availability. When grown on an organic nitrogen source, E. coli cells released NH(4)(+) and were enriched in (15)N relative to the nitrogen source (1.6-3.1 per thousand). However, when cells were grown on an inorganic nitrogen source, the biomass was depleted (6.1-9.1 per thousand) relative to the source. By measuring (15)N enrichment of microorganisms relative to nitrogen pools, ecosystem ecologists may be able to determine if microorganisms are assimilating or mineralizing nitrogen. PMID- 18399996 TI - In vivo expression technology (IVET) selection of genes of Rhizobium leguminosarum biovar viciae A34 expressed in the rhizosphere. AB - IVET was used to identify genes that are specifically expressed in the rhizosphere of the pea-nodulating bacterium Rhizobium leguminosarum A34. A library of R. leguminosarum A34 cloned in the integration vector pIE1, with inserts upstream of a promoter-less purN:gfp:gusA, was conjugated into purN host RU2249 and recombined into the genome. After removal of colonies that expressed the reporter genes of the vector under laboratory conditions, the library was inoculated into a nonsterile pea rhizosphere. The key result is that 29 rhizosphere-induced loci were identified. Sequence analysis of these clones showed that a wide variety of R. leguminosarum A34 genes are expressed specifically in the rhizosphere including those encoding proteins involved in environmental sensing, control of gene expression, metabolic reactions and membrane transport. These genes are likely to be important for survival and colonization of the pea rhizosphere. PMID- 18399997 TI - Interactions of some common pathogenic bacteria with Acanthamoeba polyphaga. AB - Protozoan grazing is a major trophic pathway whereby the biomass re-enters the food web. Nonetheless, not all bacteria are digested by protozoa and the number known to evade digestion, resulting in their environmental augmentation, is increasing. We investigated the interactions of Bacillus cereus, Enterococcus faecalis, Enteropathogenic Escherichia coli (EPEC), Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and methicillin-sensitive Staphylococcus aureus (MSSA), with the amoeba, Acanthamoeba polyphaga. There was evidence of predation of all bacterial species except L. monocytogenes and S. aureus, where extracellular numbers were significantly higher when cultured with amoebae compared with growth in the absence of amoebae. Intracellular growth kinetic experiments and fluorescent confocal microscopy suggest that S. aureus survived and may even multiply within A. polyphaga, whereas there was no apparent intra amoebal replication of L. monocytogenes and higher numbers were likely sustained on metabolic waste products released during coculture. PMID- 18399998 TI - The SOS response affects thermoregulation of colicin K synthesis. AB - Temperature is one of the key environmental parameters affecting bacterial gene expression. This study investigated the effect of temperature on synthesis of Escherichia coli colicins E1, K, N and E7 as well as the molecular basis underlying thermoregulation of the colicin K activity gene cka. The results of our study show that synthesis of the investigated colicins is higher at 37 degrees C than at 22 degrees C and that temperature regulates cka expression at the level of transcription. We propose that the SOS response indirectly regulates thermoregulation of colicin K (and possibly of the other examined colicins). Two LexA dimers bind cooperatively with high affinity to the two overlapping LexA boxes in a temperature-independent manner. At 22 degrees C the relative degree of repression is higher as a result of less LexA cleavage due to a slower growth rate, while at 37 degrees C the extent of LexA cleavage is higher due to a higher growth rate. Thermoregulation of colicin synthesis is an additional example of the connection between the SOS regulon and cell physiology. PMID- 18399999 TI - Gene cloning and characterization of a second alanine racemase from Bacillus subtilis encoded by yncD. AB - Alanine racemase activity was investigated in Bacillus subtilis. A putative second alanine racemase gene (yncD) was cloned in parallel with the previously identified alanine racemase gene, dal. Each of the B. subtilis genes, dal and yncD complemented the Escherichia coli Alr- DadX- double mutant alanine auxotrophic strain MB2159 in vivo, restoring the prototrophic phenotype. Alanine racemase activity was also detected in vitro in cell-free extracts prepared from cultures of E. coli MB2159 harboring plasmids expressing either of the cloned B. subtilis genes and preliminary characterization of enzyme activity is presented. PMID- 18400000 TI - Prevotella intermedia induces matrix metalloproteinase-9 expression in human periodontal ligament cells. AB - Matrix metalloproteinases (MMPs) play pivotal roles in inflammatory diseases including chronic periodontitis. The effects of Prevotella intermedia, a major periodontal pathogen, on MMP-9 production in primary human periodontal ligament (hPDL) cells were examined in the present study. MMP-9 mRNA expression was measured by semiquantitative reverse transcriptase PCR and its protein secretion was assayed by gelatin zymography. Prevotella intermedia ATCC 25611 supernatant time and dose-dependently induced MMP-9 expression. In contrast, Porphyromanas gingivalis ATCC 33277 supernatants, Escherichia coli lipopolysacchride and IL 1beta exhibited no stimulatory effects on MMP-9 production in hPDL cells. Mitogen activated protein kinases [MAPK, including extracellular signal-related kinases (ERK), c-jun N-terminal kinases (JNK) and p38] inhibitors exerted no effect on the P. intermedia-induced MMP-9 production, indicating that P. intermedia induced MMP-9 production through an MAPK-independent pathway. Our results demonstrated that P. intermedia may contribute to periodontal tissue destruction during chronic periodontitis by inducing MMP-9 production in hPDL cells. PMID- 18400001 TI - Causes of constraints to patient flow in emergency departments: a comparison between staff perceptions and findings from the Patient Flow Study. AB - OBJECTIVES: The objectives of the study were: (i) to survey staff perceptions of causes of delay in patients' journeys through the ED; (ii) to identify and analyse key constraints to patient flow using real-time diagnostic/decision support software (Patient Flow Study); and (iii) to assess the correlation between staff perceptions and data from the Patient Flow Study. METHODS: ED and non-ED staff were surveyed prior to the Patient Flow Study. The survey involved ranking the likely reasons for delay at three set points after triage (160, 320 and 480 min). Real-time data on delay in patients' journeys through the ED were collected over a period of 5 weeks. The correlation between staff perceptions and study data at the three time points was calculated using Spearman's rank correlation coefficient. Subgroup analysis was performed on the basis of staff position, years of experience at St George and whether they had previously attended training on constraints to flow. RESULTS: A total of 68 staff responded to the questionnaire (response rate 42%). During the study period, 4555 ED attendances (97% of all presentations) were analysed for causes of constraint. Strong correlation between staff perceptions and real-time data was only found among some subgroups at the point 160 min from triage. CONCLUSIONS: Overall, staff perceptions regarding causes of constraint to patient flow do not correlate well with data obtained from real-time analysis. PMID- 18400002 TI - Review article: Non-invasive assessment of cardiac output with portable continuous-wave Doppler ultrasound. AB - Cardiac output is considered an important parameter when assessing the cardiovascular status of a critically ill patient. Both non-invasive (e.g. bioimpedance, echocardiography) and invasive methods (Swan Ganz catheter) have been used to measure cardiac output. The ultrasonic cardiac output monitoring device provides a new method of non-invasively assessing cardiac output in various clinical settings. The ultrasonic cardiac output monitoring device was introduced clinically in 2001, and appears to be a promising adjunct in the assessment of the cardiovascular state in a variety of patient cohorts. In this short review article, we will introduce this new technique, discuss the required skills and compare it with methods already in use. In particular, a critical comparison with the 'gold standard', the invasive measurement of cardiac output with the pulmonary artery catheter, will be given. PMID- 18400003 TI - Assessing the impact of streaming in a regional emergency department. AB - OBJECTIVE: To evaluate the impact of a streaming model, previously validated in metropolitan EDs, on selected performance indicators in a regional ED. METHOD: Multiple linear regression models were applied to monthly time series data from 43 months prior to the intervention and 15 months following the intervention to measure the impact of the streaming model on the following performance indicators: (i) percentage of emergency patients admitted to an inpatient bed within 8 h; (ii) percentage of non-admitted emergency patients with a length of stay of less than 4 h; and (iii) percentage of emergency patients who left without being seen by a doctor or nurse practitioner. SETTING: Bendigo Health ED in regional Victoria. RESULTS: Prior to the introduction of streaming, there was a downward trend in both the percentage of emergency patients admitted to an inpatient bed within 8 h, and the percentage of non-admitted emergency patients with a length of stay of less than 4 h. After the introduction of streaming, these trends were reversed (P = 0.008 and P = 0.004, respectively). There was no statistically significant change in the trend associated with the percentage of emergency patients who left without being seen (P = 0.904). CONCLUSIONS: The implementation of the streaming model had an impact on the two performance indicators associated with length of stay in this regional ED, but did not have a significant impact (positive or negative) on the percentage of patients who did not wait to be seen. These results might interest other EDs in regional hospitals. PMID- 18400004 TI - Synergistic interactions between the ACC deaminase-producing bacterium Pseudomonas putida UW4 and the AM fungus Gigaspora rosea positively affect cucumber plant growth. AB - Bacteria producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase modulate plant ethylene levels. Decreased ethylene levels increase plant tolerance to environmental stresses and promote legume nodulation. On the contrary, the role of ethylene in mycorrhizal symbiosis establishment is still controversial. In this work, the ACC deaminase-producing strain Pseudomonas putida UW4 AcdS+ and its mutant AcdS(-), impaired in ACC deaminase synthesis, were inoculated alone or in combination with the AM fungus Gigaspora rosea on cucumber. Mycorrhizal and bacterial colonization as well as plant growth and morphometric parameters were measured. The influence of each microorganism on the photosynthetic efficiency was evaluated on the second and fourth leaf. The strain AcdS+, but not the AcdS( ) mutant, increased AM colonization and arbuscule abundance. The mycorrhizal fungus, but not the bacterial strains, promoted plant growth. However, the AcdS+ strain, inoculated with G. rosea, induced synergistic effects on plant biomass, total root length and total leaf projected area. Finally, the photosynthetic performance index was increased by the strain UW4 AcdS+ inoculated in combination with G. rosea BEG9. These results suggest a key role of this enzyme in the establishment and development of AM symbiosis. PMID- 18400005 TI - Ericoid mycorrhizal fungi are common root inhabitants of non-Ericaceae plants in a south-eastern Australian sclerophyll forest. AB - Fungi were isolated from the roots of 17 plant species from the families Apiaceae, Cunoniaceae, Cyperaceae, Droseraceae, Fabaceae-Mimosoideae, Lomandraceae, Myrtaceae, Pittosporaceae, Proteaceae and Stylidiaceae at a sclerophyll forest site in New South Wales, Australia. Internal transcribed spacer (ITS) restriction fragment length polymorphism (RFLP) and sequence comparisons indicated that the isolated fungi had affinities to a range of ascomycetes, basidiomycetes and zygomycetes. Four RFLP types had closest affinities to previously identified Helotiales ericoid mycorrhizal (ERM) or Oidiodendron spp. Isolates representing six RFLP types, which were variously isolated from all 17 plant species, formed ERM coils in hair root epidermal cells of Woollsia pungens (Ericaceae) under gnotobiotic conditions. Three of these isolates formed intercellular hyphae, intracellular hyphae and/or microsclerotia, which are typical of dark septate endophyte infection, in roots of Stylidium productum (Stylidiaceae), indicating an ability to form different types of association with roots of different hosts. Overall the data indicate that a broad range of plant taxa may act as repositories for ERM fungi in sclerophyll forest soil. PMID- 18400006 TI - Degradation of N-acyl homoserine lactone quorum sensing signal molecules by forest root-associated fungi. AB - A collection of mycorrhizal and nonmycorrhizal root-associated fungi coming from forest environments was screened for their ability to degrade N-acyl homoserine lactones (AHL) or to prevent AHL recognition by producing quorum sensing inhibitors (QSI). No production of QS-inhibitors or -activators was detected using the two biosensors Chromobacterium violaceum CV026 and Agrobacterium tumefaciens in the culture supernatant of these fungi. However, the ability to degrade C6- and 3O,C6-HSL was detected for three fungal isolates. Acidification assay revealed that the AHL were degraded by a lactonase activity for two of these isolates. These results demonstrated for the first time that the forest root-associated fungi are capable of degrading the AHL signal molecules. PMID- 18400007 TI - Physical organization and phylogenetic analysis of acdR as leucine-responsive regulator of the 1-aminocyclopropane-1-carboxylate deaminase gene acdS in phytobeneficial Azospirillum lipoferum 4B and other Proteobacteria. AB - The phytostimulatory alphaproteobacterium Azospirillum lipoferum 4B exhibits the plant-beneficial gene acdS, which enables deamination of the ethylene precursor 1 aminocyclopropane-1-carboxylic acid (ACC). Here, we show that acdS is in the vicinity of acdR, a homolog to leucine-responsive regulator lrp, in A. lipoferum 4B and most other acdS+ Proteobacteria. Unlike in Beta- and Gammaproteobacteria, acdS (and acdR) is preferentially located on symbiotic islands and plasmids in Alphaproteobacteria. In A. lipoferum 4B, acdS was mapped on a 750-kb plasmid that is lost during phenotypic variation, whereas other phytobeneficial genes such as nifH (associative nitrogen fixation) are maintained. In Proteobacteria, the phylogenies of acdR and acdS were largely but not totally congruent, despite physical proximity of the genes, regardless of whether DNA or deduced protein sequences were used. Potential Lrp, cAMP receptor protein (CRP) and fumarate nitrate reduction regulator (FNR) binding sites were evidenced in the acdS promoter regions of strain 4B and most of 46 other acdS+ Proteobacteria. Indeed, transcriptional and enzymatic analyses done in vitro pointed to the involvement of Lrp- and FNR-like transcriptional up-regulation of ACC deaminase activity in A. lipoferum 4B. This is the first synteny, phylogenetic, and functional analysis of factors modulating acdS expression in Azospirillum plant growth-promoting rhizobacterium. PMID- 18400008 TI - Involvement of extracellular matrix proteins in the course of experimental paracoccidioidomycosis. AB - We aimed at determining involvement of extracellular matrix proteins (ECMp) and an ECM-binding adhesin (32-kDa protein) from Paracoccidioides brasiliensis, in the course of experimental paracoccidioidomycosis. BALB/c mice were infected with P. brasiliensis conidia previously incubated with soluble laminin, fibronectin and fibrinogen or a mAb against the fungal adhesin. Inflammatory response, chitin levels and cytokine production at different postinfection periods were determined. Chitin was significantly decreased in lungs of mice infected with ECMp-treated conidia when compared with controls at week 8, especially with laminin and fibrinogen. Contrariwise, when animals were infected with mAb-treated conidia no differences in chitin content were found. The observed inflammatory reaction in lungs was equivalent in all cases. IFN-gamma increased significantly in lungs from mice infected with soluble ECMp - (at day 4 and week 12) or mAb treated conidia (at week 12) when compared with animals infected with untreated conidia. Significant increased levels of tumour necrosis factor-alpha were observed at 8 weeks in animals infected with ECMp-treated conidia while no differences were observed during the remaining periods. These findings point toward an inhibitory effect of ECMp on P. brasiliensis conidia infectivity and suggest that these proteins may interfere with conidia initial adhesion to host tissues probably modulating the immune response in paracoccidioidomycosis. PMID- 18400009 TI - Fine-epitope mapping of an antibody that binds the ectodomain of influenza matrix protein 2. AB - Our previous work found that the monoclonal antibody 8C6, which recognized the epitope EVETPIRN on influenza A virus M2 protein, conferred protection against influenza virus challenge. In this study, 8C6 was used to screen the 7-mer phage peptide library in order to identify the crucial amino acid residues on the protective epitope EVETPIRN. Nine positive phage clones were selected by a test of dose-dependent binding activity to 8C6 after three rounds of panning. The phage clones exhibited a consensus motif (TXXR), which was found on the epitope EVETPIRN. Site-directed mutation analysis indicated that Thr and Arg on the epitope EVETPIRN played a key role in the recognition by 8C6. Furthermore, sequence alignment and analysis revealed that Thr and Arg on the epitope were highly conserved. Our results could provide useful information for influenza vaccine design based on M2 mimotope. PMID- 18400010 TI - Respiratory infection with Francisella novicida induces rapid dystrophic cardiac calcinosis (DCC). AB - Francisella tularensis causes pulmonary tularemia and death in humans when left untreated. Here, using a novel aerosol infection model, we show that acute pulmonary Francisella novicida infection not only causes pneumonia and liver damage, but also induces dystrophic cardiac calcinosis (DCC) in BALB/c mice. C57BL/6 mice also develop pneumonia and hepatic damage, but fail to develop DCC. Development of DCC in BALB/c mice is associated with significant induction of RANKL but not osteopontin in their organs. Depletion of lung macrophages prior to infection markedly reduces pericarditis and calcification in BALB/c mice but does not increase their susceptibility to infection. PMID- 18400011 TI - Apoptotic neutrophils containing Staphylococcus epidermidis stimulate macrophages to release the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-6. AB - Staphylococcus epidermidis infections are usually nosocomial and involve colonization of biomaterials. The immune defense system cannot efficiently control the bacteria during these infections, which often results in protracted chronic inflammation, in which a key event is disturbed removal of neutrophils by tissue macrophages. While ingesting uninfected apoptotic neutrophils, macrophages release anti-inflammatory cytokines that lead to resolution of inflammation. In clinical studies, we have previously found elevated levels of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 in synovial fluid from prostheses infected with coagulase negative staphylococci. We show that macrophages phagocytosing apoptotic neutrophils containing S. epidermidis released TNF-alpha and interleukin-6, whereas macrophages phagocytosing spontaneously apoptotic neutrophils did not. This difference was not due to dissimilar phagocytic capacities, because macrophages ingested both types of neutrophils to the same extent. The activation was induced mainly by the apoptotic neutrophils themselves, not by the few remaining extracellular bacteria. Macrophages were not activated by apoptotic neutrophils that contained paraformaldehyde-killed S. epidermidis. Proinflammatory reactions induced by clearance of apoptotic neutrophils containing S. epidermidis might represent an important mechanism to combat the infective agent. This activation of macrophages may contribute to the development of chronic inflammation instead of inflammation resolution. PMID- 18400012 TI - Evading the host immune response: how foot-and-mouth disease virus has become an effective pathogen. AB - Foot-and-mouth disease virus (FMDV) causes an economically devastating disease of cloven-hoofed animals. In this review, we discuss the mechanisms FMDV has evolved to counteract the host innate and adaptive immune responses and the role of viral proteins in this process. The viral leader proteinase, L pro, limits the host innate response by inhibiting the induction of interferon beta (IFN beta) mRNA and blocking host cell translation. A second viral proteinase, 3C pro, may affect host cell transcription because it cleaves histone H3. Viral protein 2B in conjunction with 2C or their precursor 2BC inhibits protein trafficking through the endoplasmic reticulum and Golgi apparatus. A decrease in surface expression of major histocompatibility class I molecules during FMDV infection suggests that 2B, 2C and/or 2BC may be involved in delaying the initiation of the host adaptive immune response and also adversely affect the secretion of induced signaling molecules. FMDV also causes a transient lymphopenia in swine, but the mechanism involved is not understood nor have any viral protein(s) been implicated. Furthermore, the interaction of FMDV with various cells in the immune system including lymphocytes and dendritic cells and the possible role of apoptosis and autophagy in these interactions are discussed. PMID- 18400013 TI - CD4+ T-cell activation by antigen-presenting cells infected with urease-deficient recombinant Mycobacterium bovis bacillus Calmette-Guerin. AB - We constructed a recombinant Mycobacterium bovis bacillus Calmette-Guerin (BCG Delta UT) that lacks urease, providing acidic intraphagosomal conditions to drive an effective human immune T-cell response. BCG-Delta UT-infected macrophages stimulated autologous CD4+ T cells more efficiently than parent BCG-infected macrophages. For further T-cell activation, BCG-Delta UT-infected macrophages required pretreatment with exogenous recombinant granulocyte-macrophage colony stimulating factor or costimulation with either CD40 ligand or interferon-gamma. By contrast, BCG-Delta UT-infected dendritic cells induced significant activation of naive CD4+ T cells without costimulating signals. C57BL/6 mice intradermally inoculated with BCG-Delta UT more efficiently produced memory T cells that responded to recall antigen. Therefore, the depletion of urease from BCG is useful for the activation of T cells. PMID- 18400014 TI - Arcobacter spp. possess two very short flagellins of which FlaA is essential for motility. AB - Like Campylobacter and Helicobacter spp., Arcobacter spp. possess two flagellin genes (flaA and flaB) located adjacent to each other. The aim of this study was to characterize the flagellin proteins of Arcobacter spp., because these proteins are known virulence factors in the Epsilonproteobacteria, to which these three species belong. With the exception of Arcobacter nitrofigilis, Arcobacter flagellins are almost half the size of those in other Epsilonproteobacteria. Arcobacter flagellin proteins lack a large part of the variable central region. The low homology observed among flagellins of different Arcobacter species indicates genetic heterology between the members of this genus. Unlike in other Epsilonproteobacteria, the transcription of flagellin genes is not regulated by sigma 28- or sigma 54-dependent promoters, which suggests that transcription must be regulated in a different way in Arcobacter spp. Mutational studies revealed that only FlaA is needed for the motility of Arcobacter spp. Quantitative PCR analysis showed that transcription of flaB is higher at 30 degrees C than at 37 degrees C. Mutation of flaB had no effect on motility or on flaA transcription while mutation of flaA abolished motility and increased the transcription of flaB. These results underline that the genus Arcobacter is an unusual taxon in the epsilon subdivision of the Proteobacteria. PMID- 18400015 TI - Biofilm production by clinical strains of Acinetobacter baumannii isolated from patients hospitalized in two tertiary care hospitals. AB - Microbial biofilms are considered as virulence factors. During the present study, 34 clinical strains of Acinetobacter baumannii, isolated from patients hospitalized in two tertiary care hospitals, were examined for biofilm formation. These strains showed high variability in biofilm formation. Furthermore, no relation could be found between the ability of biofilm production and molecular type, carbapenem resistance, site of isolation of the clinical strains of A. baumannii and disease severity. Interestingly, in two cases an increase in biofilm formation could be detected in A. baumannii isolates cultured from the same patient upon prolonged hospitalization. PMID- 18400016 TI - Plant behaviour and communication. AB - Plant behaviours are defined as rapid morphological or physiological responses to events, relative to the lifetime of an individual. Since Darwin, biologists have been aware that plants behave but it has been an underappreciated phenomenon. The best studied plant behaviours involve foraging for light, nutrients, and water by placing organs where they can most efficiently harvest these resources. Plants also adjust many reproductive and defensive traits in response to environmental heterogeneity in space and time. Many plant behaviours rely on iterative active meristems that allow plants to rapidly transform into many different forms. Because of this modular construction, many plant responses are localized although the degree of integration within whole plants is not well understood. Plant behaviours have been characterized as simpler than those of animals. Recent findings challenge this notion by revealing high levels of sophistication previously thought to be within the sole domain of animal behaviour. Plants anticipate future conditions by accurately perceiving and responding to reliable environmental cues. Plants exhibit memory, altering their behaviours depending upon their previous experiences or the experiences of their parents. Plants communicate with other plants, herbivores and mutualists. They emit cues that cause predictable reactions in other organisms and respond to such cues themselves. Plants exhibit many of the same behaviours as animals even though they lack central nervous systems. Both plants and animals have faced spatially and temporally heterogeneous environments and both have evolved plastic response systems. PMID- 18400017 TI - Are there general mechanisms of animal home range behaviour? A review and prospects for future research. AB - Home range behaviour is a common pattern of space use, having fundamental consequences for ecological processes. However, a general mechanistic explanation is still lacking. Research is split into three separate areas of inquiry - movement models based on random walks, individual-based models based on optimal foraging theory, and a statistical modelling approach - which have developed without much productive contact. Here we review recent advances in modelling home range behaviour, focusing particularly on the problem of identifying mechanisms that lead to the emergence of stable home ranges from unbounded movement paths. We discuss the issue of spatiotemporal scale, which is rarely considered in modelling studies, as well as highlighting the need to consider more closely the dynamical nature of home ranges. Recent methodological and theoretical advances may soon lead to a unified approach, however, conceptually unifying our understanding of linkages among home range behaviour and ecological or evolutionary processes. PMID- 18400018 TI - Ecological consequences of genetic diversity. AB - Understanding the ecological consequences of biodiversity is a fundamental challenge. Research on a key component of biodiversity, genetic diversity, has traditionally focused on its importance in evolutionary processes, but classical studies in evolutionary biology, agronomy and conservation biology indicate that genetic diversity might also have important ecological effects. Our review of the literature reveals significant effects of genetic diversity on ecological processes such as primary productivity, population recovery from disturbance, interspecific competition, community structure, and fluxes of energy and nutrients. Thus, genetic diversity can have important ecological consequences at the population, community and ecosystem levels, and in some cases the effects are comparable in magnitude to the effects of species diversity. However, it is not clear how widely these results apply in nature, as studies to date have been biased towards manipulations of plant clonal diversity, and little is known about the relative importance of genetic diversity vs. other factors that influence ecological processes of interest. Future studies should focus not only on documenting the presence of genetic diversity effects but also on identifying underlying mechanisms and predicting when such effects are likely to occur in nature. PMID- 18400019 TI - Darwin's naturalization conundrum: dissecting taxonomic patterns of species invasions. AB - Darwin acknowledged contrasting, plausible arguments for how species invasions are influenced by phylogenetic relatedness to the native community. These contrasting arguments persist today without clear resolution. Using data on the naturalization and abundance of exotic plants in the Auckland region, we show how different expectations can be accommodated through attention to scale, assumptions about niche overlap, and stage of invasion. Probability of naturalization was positively related to the number of native species in a genus but negatively related to native congener abundance, suggesting the importance of both niche availability and biotic resistance. Once naturalized, however, exotic abundance was not related to the number of native congeners, but positively related to native congener abundance. Changing the scale of analysis altered this outcome: within habitats exotic abundance was negatively related to native congener abundance, implying that native and exotic species respond similarly to broad scale environmental variation across habitats, with biotic resistance occurring within habitats. PMID- 18400020 TI - Light chain multiple myeloma with cutaneous AL amyloidosis. AB - Cutaneous AL amyloidosis is one complication of multiple myeloma. In our patient, painful sclerotic skin changes on the extremities and macroglossia were the presenting features which led to a more detailed investigation and the diagnosis of multiple myeloma. Histological examination revealed cutaneous deposits of amyloid which were positive with Congo red stain and had an apple green color in polarized light. PMID- 18400021 TI - E-learning in the dermatological education at the Charite: evaluation of the last three years. AB - BACKGROUND: Numerous e-learning initiatives have been launched during the last decade. Many of them have not been continued, due to lack of acceptance on the part of the students, low quality or insufficient financial funding. Since 2002, the DEJAVU project has been integrated into the curriculum at the Department of Dermatology at Charite-Universitatsmedizin Berlin. DEJAVU offers an online archive of recorded lectures, lecture hand outs, structured learning modules, and case reports as well as online information about the courses/classes. METHODS: Since beginning of the summer semester 2005, the student's acceptance and utilization of the online offerings was regularly surveyed, using anonymous questionnaires handed out together with the final exams. The teaching staff's opinions about e-learning were surveyed by means of a single anonymous questionnaire. RESULTS: At the end of winter semester 2006/2007, 93.5% of the students were aware of the existence of the e-learning program. The average amount of time spent with the program was 14.7 hours over the course of one semester. 66.8% of the students considered the program as very useful for their dermatology training. The lecture notes were the most frequently used online resource. Among the teaching staff, 86% considered e-learning a useful addition to traditional teaching. CONCLUSIONS: Our results show that e-learning is very well accepted by our students. It offers an additional way of acquiring knowledge and should be used to complement traditional ways of teaching. PMID- 18400022 TI - Photodynamic therapy in dermatology--an update 2008. AB - SUMMARY: Photodynamic therapy (PDT) is used for the prevention and treatment of non-melanoma skin cancer. Until recently, clinically approved indications have been restricted to actinic keratoses, nodular and superficial basal cell carcinoma, and--since 2006--Bowen disease. However, the range of indications has been expanding continuously. PDT is also used for the treatment of non-malignant conditions such as acne vulgaris and leishmaniasis, as well as for treating premature skin aging due to sun exposure. Here, PDT is used for the stimulation of immunomodulatory effects in contrast to the induction of necrosis and apoptosis as produced in the treatment of skin tumors. The porphyrin precursor 5 aminolevulinic acid (ALA) or its methyl ester (MAL, so far the only approved formulation in Europe) is applied topically as photosensitizer to exclude systemic reactions. Possible light sources include lasers as well as incoherent light sources; irradiation with incoherent light sources is cheaper and more appropriate for large treatment areas. The main advantages of PDT in comparison to other treatment modalities are its excellent cosmetic results and its high remission rates despite low invasiveness.This article provides up-to-date information about PDT with focus on recently published studies. PMID- 18400023 TI - Does climate change affect the incidence of skin and allergic diseases in Germany? AB - BACKGROUND: Skin diseases have great socio-economic importance in Germany due to their high and in some cases still-increasing prevalence. Little attention has yet been paid to the influence of the change in climate on these diseases. OBJECTIVE: Clarify the evidence of the effects of climate change on the prevalence of skin diseases and allergies in Germany. METHODS: First, a theoretical model of the possible mechanisms and influence factors of climate and weather was created for different disease groups (skin malignancies, allergies, skin infections). Then, a systematic online and manual literature search was made for model-derived key words.The relevant publications were selected and evaluated according to a priori criteria. RESULTS: From a total of n = 31,221 hits, n = 320 publications remained for evaluation.Changes in the following parameters can be regarded as essential climatologic factors influencing the prevalence of skin and allergic diseases: temperature, UV radiation, precipitation/humidity, cloudiness, and general weather conditions.There were only a few original articles addressed to this topic. Most of them address recurring phenomena (especially levels of airborne pollen), UV radiation or ozone (and the ozone hole). Quantitative statements, prognosis models and climate scenarios have not yet been published for Germany with respect to skin diseases. CONCLUSION: Only few scientific articles on the relationship between climate changes and the prevalence of skin diseases have been published. They do not allow a reliable statement on future developments. The outlook for changes in prevalence requires further clarification using published climate models. PMID- 18400024 TI - Polymeric black tea polyphenols inhibit mouse skin chemical carcinogenesis by decreasing cell proliferation. AB - OBJECTIVES: The aim of this study was to investigate the antitumour promoting effects and possible mechanisms of action of the most abundant polymeric black tea polyphenols (PBPs 1-5) or thearubigins, in vivo. MATERIALS AND METHODS: Effect of PBP pre-treatments on 12-O-tetradecanoylphorbol-13-acetate (TPA) promoted skin papillomas was studied in 7,12-dimethylbenz(a)anthracene initiated mice over 40 weeks. Cell proliferation and apoptosis, in epidermis of the skin, were measured using appropriate immunohistochemical staining. Mitogen-activated protein kinase signalling studies were conducted with Western blot analysis at 10, 20, 30 and 40 weeks of promotion. RESULTS: Pre-treatments with PBP fractions differentially altered latency, multiplicity and incidence of skin papillomas as compared to TPA treatments thereby exhibiting antipromoting effects. Most PBP fractions decreased TPA-induced cell proliferation by decreasing activation of signalling kinases (c-Jun N-terminal protein kinase, extracellular signal regulated protein kinase, p38 protein kinase and Akt), transcription factors (activator protein-1 and nuclear factor kappa B) and inflammatory protein (cyclooxygenase 2). TPA-induced epidermal cell apoptosis was also decreased by pre-treatment with most PBP fractions. Higher levels of p53 and p21 in skin cells pre-treated with PBP fractions followed by TPA treatment as compared to only TPA treated animals suggested possible activation of a cell cycle checkpoint. CONCLUSIONS: PBP-2 was observed to be the most potent polymeric polyphenol fraction and PBP-4 and PBP-5 showed only marginal activity, whereas PBP-1 and PBP 3 displayed intermediate efficacies. In conclusion, the protective effects of PBP fractions could be attributed to inhibition of TPA-induced cellular proliferation. PMID- 18400025 TI - Bleeding on probing differentially relates to bacterial profiles: the Oral Infections and Vascular Disease Epidemiology Study. AB - AIM: Various bacterial species are differentially prevalent in periodontal health, gingivitis or periodontitis. We tested the independent associations between three bacterial groupings and gingival inflammation in an epidemiological study. MATERIAL AND METHODS: In 706 Oral Infections and Vascular Disease Epidemiology Study (INVEST) participants > or =55 years, bleeding on probing (BoP), pocket depth (PD) and subgingival plaque samples (n=4866) were assessed in eight sites per mouth. Eleven bacterial species were quantitatively assayed and grouped as follows: (i) aetiologic burden (EB, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia); (ii) putative burden (PB, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Micromonas micros, Prevotella intermedia); (iii) health associated burden (HAB, Actinomyces naeslundii, Veillonella parvula). RESULTS: After mutual adjustment for EB, PB and HAB, the BoP prevalence increased by 45% ( p<0.0001) across increasing quartiles of EB while BoP decreased by 13% ( p<0.0001) across increasing quartiles of HAB. Mean PD increased 0.8 mm and decreased 0.3 mm from the first to fourth quartiles of EB (p<0.0001) and HAB ( p<0.0001), respectively. Among 1214 plaque samples with fourth quartile EB, 60% were collected from sites with PD < or =3 mm. CONCLUSION: Bacterial species believed to be aetiologically related to periodontitis were associated with BoP in sites with minimal PD and/or attachment level (AL). Species presumed to be associated with periodontal health demonstrated inverse associations with BoP. PMID- 18400026 TI - Prevalence of periodontitis and DMFT index in patients with Crohn's disease and ulcerative colitis. AB - AIM: To compare the prevalence of periodontal disease and the decayed, missing and filled teeth (DMFT) index in patients with Crohn's disease (CD) and ulcerative colitis (UC) with those without these diseases. MATERIAL AND METHODS: Ninety-nine CD (39.0 SD+/-12.9 years), 80 UC (43.3 SD+/-13.2) and 74 healthy controls (40.3 SD+/-12.9) were compared for DMFT index and presence of periodontitis. Probing pocket depth (PPD), clinical attachment loss (CAL), bleeding on probing (BOP), plaque and DMFT index were measured on all subjects. The presence of periodontitis was defined as having CAL > or =3 mm in at least four sites in different teeth. RESULTS: Significantly more patients with UC (90.0%; p<0.001) and CD (81.8%; p=0.03) had periodontitis than controls (67.6%). Among smokers, UC patients had significantly more periodontitis. CD had a greater mean DMFT score (18.7 versus 13.9; p=0.031) compared with controls and UC had greater median PPD (2.2 versus 1.7 mm; p<0.0001) than controls. Among non smokers, CD (2.4 mm; p<0.0001) and UC showed deeper pockets (2.3 mm; p<0.0001) compared with controls (1.5 mm). UC had a greater mean DMFT score (15.3 versus 12.1; p=0.037) compared with controls. CONCLUSIONS: CD and UC patients had higher DMFT and prevalence of periodontitis than controls, but smoking was an effect modifier. PMID- 18400027 TI - Severe periodontitis in young adults is associated with sub-clinical atherosclerosis. AB - AIM: The aim of this study was to evaluate the association between severe periodontitis and sub-clinical atherosclerosis in young (< or =40 years) systemically healthy individuals. MATERIAL AND METHODS: Ninety systemically healthy subjects, 45 affected by severe periodontitis (mean age 36.35+/-3.65 years) and 45 controls without a history of periodontal disease (mean age 33.78+/ 3.28 years), were enrolled in this study. Patients and controls were paired for age, gender, body mass index and smoking habits. Carotid intima-media thickness (IMT) was bilaterally assessed by ultrasonography at the level of common carotid artery. Traditional cardiovascular risk factors for atherosclerosis were also evaluated. RESULTS: The overall mean carotid IMT was 0.82+/-0.13 mm in the test group and 0.72+/-0.07 mm in the control group ( p<0.0001). Stepwise regression analysis showed that periodontitis ( p<0.0001) and regular physical activity ( p=0.0009) were predictor variables of overall mean carotid IMT. When considering an IMT> or =0.82 mm as the critical index of increased cardiovascular risk, periodontal patients overcame this threshold compared with healthy patients by an odds ratio=8.55 [confidence interval 95%: 2.38; 39.81]. No investigated haemostatic variable was associated with increased carotid IMT. CONCLUSION: Severe periodontitis is associated with sub-clinical atherosclerosis in young systemically healthy patients. PMID- 18400028 TI - Periodontal parameters and cervical root resorption during orthodontic tooth movement. AB - OBJECTIVES: To assess the relationship between periodontal parameters and cervical root resorption in orthodontically moved teeth. MATERIAL AND METHODS: In a standardized experimental tooth movement in 16 periodontally healthy subjects, 29 pre-molars were tipped buccally for 8 weeks. Eighteen contralateral pre-molars not subjected to orthodontic movement served as controls. Plaque Index (PI), Gingival Index (GI), probing depth and bleeding on probing were assessed three times before and six times during the experimental phase. Teeth were extracted and scanned in a micro-computed tomography scanner. The presence or absence, and the severity of cervical root resorption were evaluated on the three-dimensional reconstruction of the scans by two calibrated examiners. RESULTS: Overall, periodontal parameters were not different between the test and the control teeth. Clear signs of buccal cervical resorption were detected on 27 of 29 orthodontically moved teeth and on one control tooth. Ten subjects had perfect oral hygiene and no gingivitis, whereas six subjects showed a moderate level of plaque and gingivitis (>20% occurrences of PI or GI with >0). No relationship could be demonstrated between resorption and periodontal parameters. CONCLUSIONS: Nearly all orthodontically moved teeth showed signs of cervical resorption. Periodontal parameters were unrelated to this important side effect of orthodontic treatment. PMID- 18400029 TI - In vivo degradation of nitric oxide synthase (NOS) and heat shock protein 90 (HSP90) by calpain is modulated by the formation of a NOS-HSP90 heterocomplex. AB - We have shown previously that isolated heat shock protein 90 (HSP90) and nitric oxide synthase (NOS), once associated in a heterocomplex, become completely resistant to calpain digestion. In this study, it is shown that, in vivo, under conditions of calpain activation, the protection of NOS degradation occurs. In addition, the extent of NOS degradation is a function of the level of HSP90 expression. Thus, in rat brain, which contains a large excess of HSP90, almost all neuronal NOS is associated with the chaperone protein. In this condition, neuronal NOS retains its full catalytic activity, although limited proteolytic conversion to still active low-molecular-mass (130 kDa) products takes place. In contrast, in aorta, which contains much smaller amounts of HSP90, endothelial NOS is not completely associated with the chaperone, and undergoes extensive degradation with a loss of protein and catalytic activity. On the basis of these findings, we propose a novel role of the HSP90-NOS heterocomplex in protecting in vivo NOS from proteolytic degradation by calpain. The efficiency of this effect is directly related to the level of intracellular HSP90 expression, generating a high HSP90 to NOS ratio, which favours both the formation and stabilization of the HSP90-NOS heterocomplex. This condition seems to occur in rat brain, but not in aorta, thus explaining the higher vulnerability to proteolytic degradation of endothelial NOS relative to neuronal NOS. PMID- 18400031 TI - Identification in the yeast Pichia stipitis of the first L-rhamnose-1 dehydrogenase gene. AB - There are two distinctly different pathways for the catabolism of l-rhamnose in microorganisms. One pathway with phosphorylated intermediates was described in bacteria; here the enzymes and the corresponding gene sequences are known. The other pathway has no phosphorylated intermediates and has only been described in eukaryotic microorganisms. For this pathway, the enzyme activities have been described but not the corresponding gene sequences. The first enzyme in this catabolic pathway is the NAD-utilizing L-rhamnose 1-dehydrogenase. The enzyme was purified from the yeast Pichia stipitis, and the mass of its tryptic peptides was determined using MALDI-TOF MS. This enabled the identification of the corresponding gene, RHA1. It codes for a protein with 258 amino acids belonging to the protein family of short-chain alcohol dehydrogenases. The ORF was expressed in Saccharomyces cerevisiae. As the gene contained a CUG codon that codes for serine in P. stipitis but for leucine in S. cerevisiae, this codon has changed so that the same amino acid was expressed in S. cerevisiae. The heterologous protein showed the highest activity and affinity with L-rhamnose and a lower activity and affinity with L-mannose and L-lyxose. The enzyme was specific for NAD. A northern blot analysis revealed that transcription in P. stipitis is induced during growth on L-rhamnose but not on other carbon sources. PMID- 18400030 TI - Identification and functional characterization of an aggregation domain in long myosin light chain kinase. AB - The functions of long smooth muscle myosin light chain kinase (L-MLCK), a molecule with multiple domains, are poorly understood. To examine the existence of further potentially functional domains in this molecule, we analyzed its amino acid sequence with a tango program and found a putative aggregation domain located at the 4Ig domain of the N-terminal extension. To verify its aggregation capability in vitro, expressible truncated L-MLCK variants driven by a cytomegalovirus promoter were transfected into cells. As anticipated, only the overexpression of the 4Ig fragment led to particle formation in Colon26 cells. These particles contained 4Ig polymers and actin. Analysis with detergents demonstrated that the particles shared features in common with aggregates. Thus, we conclude that the 4Ig domain has a potent aggregation ability. To further examine this aggregation domain in vivo, eight transgenic mouse lines expressing the 4Ig domain (4Ig lines) were generated. The results showed that the transgenic mice had typical aggregation in the thigh and diaphragm muscles. Histological examination showed that 7.70 +/- 1.86% of extensor digitorum longus myofibrils displayed aggregates with a 36.44% reduction in myofibril diameter, whereas 65.13 +/- 3.42% of diaphragm myofibrils displayed aggregates and the myofibril diameter was reduced by 43.08%. Electron microscopy examination suggested that the aggregates were deposited at the mitochondria, resulting in structural impairment. As a consequence, the oxygen consumption of mitochondria in the affected muscles was also reduced. Macrophenotypic analysis showed the presence of muscular degeneration characterized by a reduction in force development, faster fatigue, decreased myofibril diameters, and structural alterations. In summary, our study revealed the existence of a novel aggregation domain in L-MLCK and provided a direct link between L-MLCK and aggregation. The possible significance and mechanism underlying the aggregation-based pathological processes mediated by L-MLCK are also discussed. PMID- 18400032 TI - A novel bradykinin potentiating peptide isolated from Bothrops jararacussu venom using catallytically inactive oligopeptidase EP24.15. AB - Characterization of the peptide content of venoms has a number of potential benefits for basic research, clinical diagnosis, development of new therapeutic agents, and production of antiserum. Here, we use a substrate-capture assay that employs a catalytically inactive mutant of thimet oligopeptidase (EC 3.4.24.15; EP24.15) to identify novel bioactive peptides in Bothrops jararacussu venom. Of the peptides captured with inactive EP24.15 and identified by mass spectrometry, three were previously identified bradykinin-potentiating peptides (BPP), 1000 colonoscopies) did not increase overall accuracy. Less experienced endoscopists had an increased accuracy rate with dye-spraying (76.7%vs 59.3%, P < 0.05) but experienced endoscopists had an increased accuracy rate with water insufflation (67.4%vs 63.3%, P > .05). CONCLUSION: Currently, terminal ileal imaging is not a reliable mode of documenting complete colonoscopy. Using water insufflation or dye-spraying coupled with modifications in image acquisition technique may improve its reliability but these methods require further investigation before they can replace the use of caecal landmarks as completion parameters. PMID- 18400042 TI - The BSG/ACPGBI guidelines for colonoscopic screening: what are we missing? AB - INTRODUCTION: Before publication of the British Society of Gastroenterology and Association of Coloproctology of Great Britain and Ireland guidelines in 2002, screening for people with a family history of colorectal cancer was sporadic and largely dependant on unvalidated local guidelines. Since 1990 we have been screening patients with both high and moderate risk family histories of colorectal cancer using local protocols which were more liberal than the new guidelines. In this study, we have analysed the pathology that would have been missed if we had been using the new guidelines in the period 1990-2002. METHOD: A total of 399 consecutive patients with a positive family history of colorectal malignancy underwent screening endoscopy according to local guidelines. Demographic, endoscopic and pathologic data were prospectively collected. Patients were retrospectively divided into those who would have been screened under the new guidelines (group 1) and those who would not (group 2). The recorded pathology was graded as significant or insignificant and the findings compared between the two groups. RESULTS: A total of 399 patients underwent 557 endoscopies of which 278 (50%) were indicated under the new guidelines (group 1) and 279 (50%) were not indicated (group 2). A significant pathology or carcinoma was found in 15.8% of group 1 endoscopies and 10.0% of group 2 endoscopies. This difference was significant. CONCLUSION: If we had been using the new guidelines in the period 1990-2002, we would not have performed 279 (50%) of the 557 procedures, but would not have discovered significant pathology in 10% of the moderate risk endoscopies representing 39% of the significant pathology, which was actually present in this population. PMID- 18400043 TI - Stereological assessment of the epithelial surface area of the ovine palatine and pharyngeal tonsils. AB - Although ovine tonsils play a major role in prion diseases, their morphology is poorly documented and morphometric data are sparse. Therefore, a stereological assessment of the surface area of the palatine and pharyngeal tonsils of five sheep was performed. The epithelial surface area of both tonsils is considerably increased by the presence of tonsillar crypts and folds. The mean epithelial surface area of the paired palatine tonsil was 7.14 cm(2) and that of the pharyngeal tonsil was 23.5 cm(2). In comparison with the paired palatine tonsil, the volume of the pharyngeal tonsil was almost two times larger, whilst its surface area was more than three times larger. PMID- 18400044 TI - Gastric submucosal fat in cats. AB - A previous study showed that a gastric intramural radiolucent band seen on radiographs and a gastric intramural hypoattenuating layer seen on computed tomography were due to the presence of gastric submucosal fat in a cat. There is no known prevalence and distribution pattern of this gastric submucosal fat in cats. This study was carried out to determine the prevalence and distribution pattern of submucosal fat in cat stomachs. Histologic examination of 41 cat stomachs was performed after scheduled killing of cats, obtained from animal shelters. The results revealed that submucosal fat was present in all stomachs. However, thickness and distribution pattern of the submucosal fat varied. PMID- 18400045 TI - Observations on the macroscopic anatomy of the intestinal tract and its mesenteric folds in the pampas deer (Ozotoceros bezoarticus, Linnaeus 1758). AB - We described the macroscopic anatomy of the intestines and their peritoneal folds of five adult pampas deer (Ozotoceros bezoarticus), a cervid species considered to ingest a high proportion of grass in its natural diet. The mean (+/-SD) body weight was 17 (+/-2) kg. The small intestine and the caecocolon measured 495 (+/ 37) cm and 237 (+/-24) cm in length, respectively, with an average ratio (small intestine:caecocolon) of 1.9 (+/-0.1). The ascending colon had two and a half centripetal gyri, a central flexure and two centrifugal gyri. The spiral ansa, which was similar to an ellipse, was fixed to the whole left face of the mesenterium. Apart from the peritoneal folds described in the Nomina Anatomica Veterinaria, three additional, hitherto not described folds were found: a fold that fixed the caecum to the proximal ansa of the ascending colon, one that joined the terminal part of the proximal ansa to the last centrifugal gyrus of the spiral ansa of the ascending colon, and one that linked the ascending duodenum to the proximal ansa of the ascending colon. When compared with published data from other cervids of different feeding niches, it appears that, among cervids, the ratio of small intestine to the caecocolon length does not reflect the natural diet. PMID- 18400046 TI - Anti-O6-methylguanine-methyltransferase (MGMT) immunohistochemistry in glioblastoma multiforme: observer variability and lack of association with patient survival impede its use as clinical biomarker. AB - Silencing of O6-methylguanine-DNA methyltransferase (MGMT) protein expression because of MGMT gene promoter hypermethylation is considered to be associated with postoperative chemoradiotherapy benefits in glioblastoma multiforme (GBM) patients. The objective of this study was to clarify the usability of MGMT immunohistochemistry (IHC) as a clinical biomarker. We immunostained a tissue microarray containing biopsy samples of 164 GBM patients from the European Organization for Research and Treatment of Cancer and the National Cancer Institute of Canada (EORTC/NCIC) trial 26981/22981 using two commercial anti-MGMT antibodies (clones MT3.1 and MT23.2). Immunostaining results were semiquantitatively evaluated by four observers from three neuropathological laboratories using a predefined algorithm. We analyzed (i) inter- and intraobserver agreement on MGMT expression (kappa statistics); (ii) correlation of MGMT expression with MGMT promoter methylation status (kappa statistics); and (iii) correlation of MGMT expression with patient outcome (log-rank test). Interobserver agreement on MGMT expression varied from slight to almost perfect, whereas intraobserver agreement ranged from substantial to almost perfect. MGMT expression showed poor to moderate correlation with MGMT promoter methylation status. We found no significant association of MGMT expression with patient outcome. In our hands, observer variability as well as lack of association with the MGMT promoter methylation status and patient survival impeded the use of anti MGMT immunohistochemistry as a clinical biomarker for routine diagnostic purposes. PMID- 18400047 TI - Strain-associated variations in abnormal PrP trafficking of sheep scrapie. AB - Prion diseases are associated with the accumulation of an abnormal form of the host-coded prion protein (PrP). It is postulated that different tertiary or quaternary structures of infectious PrP provide the information necessary to code for strain properties. We show here that different light microscopic types of abnormal PrP (PrP(d)) accumulation found in each of 10 sheep scrapie cases correspond ultrastructurally with abnormal endocytosis, increased endo-lysosomes, microfolding of plasma membranes, extracellular PrP(d) release and intercellular PrP(d) transfer of neurons and/or glia. The same accumulation patterns of PrP(d) and associated subcellular lesions were present in each of two scrapie strains present, but they were present in different proportions. The observations suggest that different trafficking pathways of PrP(d) are influenced by strain and cell type and that a single prion strain causes several PrP(d)-protein interactions at the cell membrane. These results imply that strains may contain or result in production of multiple isoforms of PrP(d). PMID- 18400048 TI - Smooth muscle NOS, colocalized with caveolin-1, modulates contraction in mouse small intestine. AB - Neuronal nitric oxide synthase (nNOS) in myenteric neurons is activated during peristalsis to produce nitric oxide which relaxes intestinal smooth muscle. A putative nNOS is also found in the membrane of intestinal smooth muscle cells in mouse and dog. In this study we studied the possible functions of this nNOS expressed in mouse small intestinal smooth muscle colocalized with caveolin-1(Cav 1). Cav-1 knockout mice lacked nNOS in smooth muscle and provided control tissues. 60 mM KCl was used to increase intracellular [Ca(2+)] through L-type Ca(2+) channel opening and stimulate smooth muscle NOS activity in intestinal tissue segments. An additional contractile response to LNNA (100 microM, NOS inhibitor) was observed in KCl-contracted tissues from control mice and was almost absent in tissues from Cav-1 knockout mice. Disruption of caveolae with 40 mM methyl-beta cyclodextrin in tissues from control mice led to the loss of Cav-1 and nNOS immunoreactivity from smooth muscle as shown by immunohistochemistry and a reduction in the response of these tissues to N-omega-nitro-L-arginine (LNNA). Reconstitution of membrane cholesterol using water soluble cholesterol in the depleted segments restored the immunoreactivity and the response to LNNA added after KCl. Nicardipine (1 microM) blocked the responses to KCl and LNNA confirming the role of L-type Ca(2+) channels. ODQ (1 microM, soluble guanylate cyclase inhibitor) had the same effect as inhibition of NOS following KCl. We conclude that the activation of nNOS, localized in smooth muscle caveolae, by calcium entering through L-type calcium channels triggers nitric oxide production which modulates muscle contraction by a cGMP-dependent mechanism. PMID- 18400049 TI - Porosome in astrocytes. AB - Secretion is a universal cellular process occurring in bakers yeast, to the complex multicellular organisms, to humans beings. Neurotransmission, digestion, immune response or the release of hormones occur as a result of cell secretion. Secretory defects result in numerous diseases and hence a molecular understanding of the process is critical. Cell secretion involves the transport of vesicular products from within cells to the outside. Porosomes are permanent cup-shaped supramolecular structures at the cell plasma membrane, where secretory vesicles transiently dock and transiently fuse to release intravesicular contents to the outside. In the past decade, porosomes have been determined to be the universal secretory machinery in cells, present in the exocrine pancreas, endocrine and neuroendocrine cells, and in neurons. In this study, we report for the first time the presence of porosomes in rat brain astrocytes. Using atomic force microscopy on live astrocytes, cup-shaped porosomes measuring 10-15 nm are observed at the cell plasma membrane. Further studies using electron microscopy confirm the presence of porosomes in astrocytes. Analogous to neuronal porosomes, there is a central plug in the astrocyte porosome complex. Immunoisolation and reconstitution of the astrocyte porosome in lipid membrane, demonstrates a structure similar to what is observed in live cells. These studies demonstrate that in astrocytes, the secretory apparatus at the cell plasma membrane is similar to what is found in neurons. PMID- 18400050 TI - Improved outcome of EAN, an animal model of GBS, through amelioration of peripheral and central inflammation by minocycline. AB - Experimental autoimmune neuritis (EAN) is a widely used animal model of the human acute inflammatory demyelinating polyradiculoneuropathy, which is the most common subtype of Guillain-Barre Syndrome. EAN is pathologically characterized by breakdown of the blood-nerve barrier, infiltration of reactive immune cells, local inflammation, demyelination in the peripheral nervous system and mechanical allodynia. Minocycline is known to have neuroprotective and anti-inflammatory effects. Furthermore, relieve of neuropathic pain following minocycline administration was observed in a variety of animal models. Here, we investigated the effects of minocycline on rat EAN. Suppressive treatment with minocycline (50 mg/kg body weight daily immediately after immunization) significantly attenuated the severity and duration of EAN. Macrophage and T-cell infiltration and demyelination in sciatic nerves of EAN rats treated with minocycline were significantly reduced compared to phosphate-buffered saline (PBS)-treated EAN rats. mRNA expressions of matrix metallopeptidase-9, inducible nitric oxide synthase and pro-inflammatory cytokines interleukin-1 beta and tumour necrosis factor-alpha in EAN sciatic nerves were greatly decreased by administration of minocycline as well. Furthermore, minocycline attenuated mechanical allodynia in EAN rats and greatly suppressed spinal microglial activation. All together, our data showed that minocycline could effectively suppress the peripheral and spinal inflammation (immune activation) to improve outcome in EAN rats, which suggests that minocycline may be considered as a potential candidate of pharmacological treatment for autoimmune-mediated neuropathies. PMID- 18400051 TI - The novel adipocytokine visfatin exerts direct cardioprotective effects. AB - Visfatin is an adipocytokine capable of mimicking the glucose-lowering effects of insulin and activating the pro-survival kinases phosphatidylinositol-3-OH kinase (PI3K)-protein kinase B (Akt) and mitogen-activated protein kinase kinase 1 and 2 (MEK1/2)-extracellular signal-regulated kinase 1 and 2 (Erk 1/2). Experimental studies have demonstrated that the activation of these kinases confers cardioprotection through the inhibition of the mitochondrial permeability transition pore (mPTP). Whether visfatin is capable of exerting direct cardioprotective effects through these mechanisms is unknown and is the subject of the current study. Anaesthetized C57BL/6 male mice were subjected to in situ 30 min. of regional myocardial ischaemia and 120 min. of reperfusion. The administration of an intravenous bolus of visfatin (5 x 10(-6) micromol) at the time of myocardial reperfusion reduced the myocardial infarct size from 46.1+/ 4.1% in control hearts to 27.3+/-4.0% (n>or= 6/group, P<0.05), an effect that was blocked by the PI3K inhibitor, wortmannin, and the MEK1/2 inhibitor, U0126 (48.8+/-5.5% and 45.9+/-8.4%, respectively, versus 27.3+/-4.0% with visfatin; n>or= 6/group, P<0.05). In murine ventricular cardiomyocytes subjected to 30 min. of hypoxia followed by 30 min. of reoxygenation, visfatin (100 ng/ml), administered at the time of reoxygenation, reduced the cell death from 65.2+/ 4.6% in control to 49.2+/-3.7%(n>200 cells/group, P<0.05), an effect that was abrogated by wortmannin and U0126 (68.1+/-5.2% and 59.7+/-6.2%, respectively; n>200 cells/group, P>0.05). Finally, the treatment of murine ventricular cardiomyocytes with visfatin (100 ng/ml) delayed the opening of the mPTP induced by oxidative stress from 81.2+/-4 sec. in control to 120+/-7 sec. (n>20 cells/group, P<0.05) in a PI3K- and MEK1/2-dependent manner. We report that the adipocytokine, visfatin, is capable of reducing myocardial injury when administered at the time of myocardial reperfusion in both the in situ murine heart and the isolated murine cardiomyocytes. The mechanism appears to involve the PI3K and MEK1/2 pathways and the mPTP. PMID- 18400054 TI - Could glucose be a proaging factor? AB - There is an ever-increasing scientific interest for the interplay between cell's environment and the aging process. Although it is known that calorie restriction affects longevity, the exact molecular mechanisms through which nutrients influence various cell signalling/modulators of lifespan remain a largely unresolved issue. Among nutrients, glucose constitutes an evolutionarily stable, precious metabolic fuel, which is catabolized through glycolytic pathway providing energy in the form of ATP and consuming NAD. Accumulating evidence shows that among the important regulators of aging process are autophagy, sirtuin activity and oxidative stress. In light of recent work indicating that glucose availability decreases lifespan whilst impaired glucose metabolism extends life expectancy, the present article deals with the potential role of glucose in the aging process by regulating--directly through its metabolism or indirectly through insulin secretion--autophagy, sirtuins as well as other modulators of aging like oxidative stress and advanced glycation end-products (AGEs). PMID- 18400052 TI - Caveolin-1: an ambiguous partner in cell signalling and cancer. AB - Caveolae are small plasma membrane invaginations that have been implicated in a variety of functions including transcytosis, potocytosis and cholesterol transport and signal transduction. The major protein component of this compartment is a family of proteins called caveolins. Experimental data obtained in knockout mice have provided unequivocal evidence for a requirement of caveolins to generate morphologically detectable caveolae structures. However, expression of caveolins is not sufficient per seto assure the presence of these structures. With respect to other roles attributed to caveolins in the regulation of cellular function, insights are even less clear. Here we will consider, more specifically, the data concerning the ambiguous roles ascribed to caveolin-1 in signal transduction and cancer. In particular, evidence indicating that caveolin 1 function is cell context dependent will be discussed. PMID- 18400055 TI - Identification of a localized mucosal immune response in rainbow trout, Oncorhynchus mykiss (Walbaum), following immunization with a protein-hapten antigen. AB - The ability of rainbow trout, Oncorhynchus mykiss (RBT), to produce a localized mucosal immune response was investigated following intraperitoneal (i.p.) or peranal (p.a.) immunization with a protein-hapten carrier, fluorescein isothiocyanate conjugated to keyhole limpet haemocyanin (FITC/KLH). Antibody levels in serum, mucus, tissue culture supernatant from blood and spleen leucocytes, and excised skin, intestine and gill tissues were determined by ELISA. Significantly, elevated antigen-specific antibodies were elicited in both serum and mucus of fish immunized i.p. Mucosal antibody responses, in general, paralleled serum responses over time. Leucocytes isolated from spleen and blood of i.p. immunized fish at week 10 produced significantly elevated antibody levels against FITC when cultured in vitro. Excised skin, intestine and gill tissues from these fish also exhibited significantly elevated antibody responses indicating localized production in the mucosa from tissue-specific B cells. A localized mucosal immune response was elicited only after i.p. and not p.a. immunization, suggesting that systemically stimulated B cells migrate to mucosal tissues where they produce antibodies locally. PMID- 18400053 TI - Emerging roles for protein histidine phosphorylation in cellular signal transduction: lessons from the islet beta-cell. AB - Protein phosphorylation represents one of the key regulatory events in physiological insulin secretion from the islet beta-cell. In this context, several classes of protein kinases (e.g. calcium-, cyclic nucleotide- and phospholipid-dependent protein kinases and tyrosine kinases) have been characterized in the beta-cell. The majority of phosphorylated amino acids identified include phosphoserine, phosphothreonine and phosphotyrosine. Protein histidine phosphorylation has been implicated in the prokaryotic and eukaryotic cellular signal transduction. Most notably, phoshohistidine accounts for 6% of total protein phosphorylation in eukaryotes, which makes it nearly 100-fold more abundant than phosphotyrosine, but less abundant than phosphoserine and phosphothreonine. However, very little is known about the number of proteins with phosphohistidines, since they are highly labile and are rapidly lost during phosphoamino acid identification under standard experimental conditions. The overall objectives of this review are to: (i) summarize the existing evidence indicating the subcellular distribution and characterization of various histidine kinases in the islet beta-cell, (ii) describe evidence for functional regulation of these kinases by agonists of insulin secretion, (iii) present a working model to implicate novel regulatory roles for histidine kinases in the receptor independent activation, by glucose, of G-proteins endogenous to the beta-cell, (iv) summarize evidence supporting the localization of protein histidine phosphatases in the islet beta-cell and (v) highlight experimental evidence suggesting potential defects in the histidine kinase signalling cascade in islets derived from the Goto-Kakizaki (GK) rat, a model for type 2 diabetes. Potential avenues for future research to further decipher regulatory roles for protein histidine phosphorylation in physiological insulin secretion are also discussed. PMID- 18400056 TI - gyrA and parC associated with quinolone resistance in Vibrio anguillarum. PMID- 18400057 TI - Editorial: prevention in the community and treatment in the clinic - two different methodological approaches to determining the evidence-base for child mental health interventions. PMID- 18400058 TI - Meta-analysis of randomized, controlled treatment trials for pediatric obsessive compulsive disorder. AB - OBJECTIVE: To conduct a meta-analysis on randomized, controlled treatment trials of pediatric obsessive-compulsive disorder (OCD). METHOD: Studies were included if they employed randomized, controlled methodology and treated young people (19 years or under) with OCD. A comprehensive literature search identified 13 RCTs containing 10 pharmacotherapy to control comparisons (N = 1016) and five cognitive-behavioral therapy (CBT) to control comparisons (N = 161). RESULTS: Random effects modeling yielded statistically significant pooled effect size (ES) estimates for pharmacotherapy (ES = .48, 95% CI = .36 to .61, p < .00001) and CBT (ES = 1.45, 95% CI = .68 to 2.22, p = .002). The results were robust to publication bias. CONCLUSIONS: This is the first meta-analysis of treatment RCTs for pediatric OCD. CBT and pharmacotherapy were the only treatments effective beyond control in alleviating OCD symptoms. CBT showed a greater ES than pharmacotherapy. Previous meta-analyses that included uncontrolled trials exaggerated the efficacy of both treatments. PMID- 18400059 TI - Heterogeneity in antisocial behaviours and comorbidity with depressed mood: a behavioural genetic approach. AB - BACKGROUND: Antisocial behaviour is often comorbid with depressed mood but is itself a collection of heterogeneous behaviours. Using a genetically informative design, we examine heterogeneity in antisocial behaviour and overlaps between different forms of antisocial behaviour with depressed mood. METHODS: Data were drawn from the G1219 large-scale community sample containing self-report questionnaire data from 941 twin pairs and 328 sibling-pairs aged 12-21 years. RESULTS: Inter-correlations among oppositionality, physical aggression and delinquency, and between the antisocial subscales and depressed mood, were mediated by genetic and non-shared environmental influences. Genetic factors generally contributed more to the correlations than the non-shared environment. CONCLUSIONS: These results indicate that many genetic and non-shared environmental influences were common to all the traits studied, but there was also evidence for trait-specific effects. PMID- 18400060 TI - Contrasting deficits on executive functions between ADHD and reading disabled children. AB - BACKGROUND: The object of this study was to analyze the executive functioning of children with attention deficit hyperactivity disorder (ADHD) or reading disability (RD) independent of their non-executive deficits. METHODS: Three carefully diagnosed groups of children, aged between 7 and 12 years (35 ADHD, 22 RD and 30 typically developing children), were tested on a wide range of tasks related to five major domains of executive functioning (EF): inhibition, visual working memory, planning, cognitive flexibility, and verbal fluency. Additional tasks were selected for each domain to control for non-executive processing. RESULTS: ADHD children were impaired on interference control, but not on prepotent and ongoing response suppression. ADHD showed deficits on visual working memory, planning, cognitive flexibility and phonetic fluency. RD children were impaired on phonetic fluency. The only EF measure that differentiated ADHD from RD was planning. CONCLUSIONS: The present sample of ADHD children showed several EF deficits, whereas RD children were almost spared executive dysfunction, but exhibited deficits in phonetic fluency. PMID- 18400061 TI - Culture and psychiatric symptoms in Puerto Rican children: longitudinal results from one ethnic group in two contexts. AB - BACKGROUND: The development of youth psychopathology may be associated with direct and continuous contact with a different culture (acculturation) and to distress related to this process (cultural stress). We examine cultural experiences of Puerto Rican families in relation to youth psychiatric symptoms in two different contexts: one in which migrant Puerto Ricans reside on the mainland as an ethnic minority and another in which they reside in their place of origin. METHODS: SAMPLE: Probability samples of 10- to 13-year-old youth of Puerto Rican background living in the South Bronx, New York City (SB) and in the San Juan Metropolitan area in Puerto Rico (PR) (N = 1,271) were followed over time. measures: Three assessments of internalizing psychiatric symptoms (elicited through the DISC-IV) and of antisocial behaviors (ASB) quantified through a six point index were carried out. Independent variables included scales of adult and child acculturation and cultural stress, and other putative correlates. DATA ANALYSIS: Within each study site, multilevel linear regression models were examined. RESULTS: Parental acculturation was associated with ASB in youth at both sites, but youth acculturation itself was not related to psychiatric symptoms. At both contexts, cultural stress was a more consistent correlate of youth psychiatric symptoms than acculturation after controlling for nativity, maternal education, child gender, stressful life events and parental psychopathology. However, the strength of the youth cultural stress association decreased over time. CONCLUSION: The association between cultural factors and child psychiatric symptoms is not restricted to contexts where an ethnic group is a minority. PMID- 18400062 TI - Human melanocytes express functional Toll-like receptor 4. AB - BACKGROUND: Melanocytes are not simply pigment-producing cells, but produce substances with a range of biological functions including antimicrobial defense. Recent studies suggest that Toll-like receptors (TLRs) play an important role in the cellular response through the recognition of pathogens. OBJECTIVES: To investigate whether TLR4 and their adapter molecules are expressed in human melanocytes. The regulation and functional role of TLR4 on cell activation were also investigated. METHODS: The expression of TLR4 in human melanocytes was determined by using reverse transcription polymerase chain reaction, western blotting, immunochemistry and flow cytometry. In vivo expression of TLR4 in melanocytes of normal human epidermis was detected by immunohistochemical double staining. The effects of gram-negative bacterial derived lipopolysaccharide (LPS) on pigmentation were investigated with the measurement of melanin content. RESULTS: TLR4 and its adaptor molecule CD14 and myeloid differentiation protein gene (MyD88) were constitutively expressed in cultured human melanocytes. Co staining of histological human skin sections with TLR4 and a melanocyte marker, gp100, confirmed the expression of TLR4 in melanocytes under physiological conditions. LPS upregulated the expression of TLR4 and MyD88 and induced NF kappaB nuclear translocation B nuclear translocation in melanocytes. Treatment of LPS increased pigmentation of human melanocytes. CONCLUSIONS: We demonstrated that functional TLR4 was expressed in human melanocytes. Our findings suggest that TLR4 may play a role in microbial-induced melanogenesis. PMID- 18400063 TI - Searching for foreign antigens as possible triggering factors of autoimmunity: Torque Teno virus DNA prevalence is elevated in sera of patients with bullous pemphigoid. AB - OBJECTIVE: The Torque Teno virus (TTV), a member of virus genus Anellovirus has been shown to be commonly present in humans, yet without detectable pathogenicity. Recent studies imply that TTV may contribute to provoke autoimmune progresses in systemic lupus erythematosus and idiopathic inflammatory myopathies. We aimed to study the presence of TTV in a group of patients with autoimmune bullous diseases with a further goal to identify long-lasting foreign antigen, such as TTV as possible triggers of skin-specific autoimmunity. PATIENTS AND METHODS: We performed in silico research to study similarities between known TTV sequences and antigens of bullous pemphigoid (BP), pemphigus vulgaris (PV) and dermatitis herpetiformis (DH). Basic Local Alignment Search Tool results showed matching regions for the major BP antigens BP180 and BP230, PV antigen desmoglein 3 and DH antigen transglutaminase 3 and disclosed overlapping, antigen predicted sequences only for BP180 regions. We also assessed the prevalence of TTV in these disorders and compared them with the results from two healthy blood donor groups (group 1: sex- and age-matched for the general bullous group, n = 95; group 2: sex- and age-matched for BP, n = 50). Furthermore, we assayed lymphocytes from four TTV DNA and BP180 NC16A blot-positive BP patients and three controls in a standard lymphocyte transformation test with a TTV peptide from the conserved ORF(Open Reading Frame)1/N22 region. RESULTS: We found that the detection rate of TTV was comparable with that in healthy controls in the group of PV (19/33); whereas detection rates in DH showed a slight, but not significant tendency for elevation (17/20). Contrary, the TTV prevalence in BP patients was significantly elevated (group 1: 36/40 vs group 2: 31/50, P < 0.032). Lymphocytes from all four virus-positive BP patients heavily reacted to TTV peptide while two of the three healthy controls have shown not to recognize the viral sequences. Only the TTV carrier healthy control had a minor reaction at lowest peptide concentration. The combined in silico, polymerse chain reaction and in vitro cell assay data of the present study indicate that a TTV persistence may contribute to the pathogenesis of BP. PMID- 18400064 TI - What causes hidradenitis suppurativa? AB - Hidradenitis suppurativa (HS)--a rather common, very chronic and debilitating inflammatory skin appendage disorder with a notoriously underestimated burden of disease--has long been a playground for the high priests of nomenclature: Ask a bunch of eminent dermatologists and skin pathologists to publicly share their thoughts on what causes HS, and they will soon get entrenched in a heated debate on whether this historical term is a despicable misnomer. Fortunately, the recently founded Hidradenitis Suppurativa Foundation (HSF; http://www.hs foundation.org), to which EXP DERMATOL serves as home journal, has broken with this unproductive tradition and has encouraged publication of the current CONTROVERSIES feature. This is exclusively devoted to discussing the pathobiology of this chronic neutrophilic folliculitis of unknown origin. Although traces of terminological bickering remain visible, it does the HS experts in our virtual debate room credit that they engage in a constructive and comprehensive dissection of potential pathogenesis pathways that may culminate in the clinical picture we know under the competing terms HS or acne inversa. These experts sketch more often complementary than mutually exclusive pathogenesis scenarios, and the outlines of a conceivable consensus on the many open pathobiology questions begin to emerge in these CONTROVERSIES. Hopefully, this heralds a welcome new tradition: to get to the molecular heart of HS pathogenesis, which can only be achieved by a renaissance of solid basic HS research, as the key to developing more effective HS therapy. PMID- 18400075 TI - The prevalence of transmitted antiretroviral drug resistance in treatment-naive patients and factors influencing first-line treatment regimen selection. AB - OBJECTIVES: To estimate the prevalence of transmitted antiretroviral (ARV) drug resistance, and to assess whether resistance testing influences first-line ARV regimen selection. METHODS: Data on patients' characteristics were collected through questionnaires. ARV drug resistance was tested by genotypic methods and defined by Quest-Stanford classification rule. Physicians reported the intended and actual treatments and the factors considered in treatment selection. RESULTS: Two hundred and twenty-eight patients were included. The prevalence of ARV drug resistance was 12.1%, with 9.8% for non-nucleoside reverse transcriptase inhibitors (NNRTIs), 4.5% for nucleoside reverse transcriptase inhibitors and 1.8% for protease inhibitors (PIs). Pill burdens, dosing frequency and physicians' experience with regimens were the major factors considered in treatment selection. The intended and actual treatment differed for 73 and 44% of the patients with and without ARV drug resistance, respectively [odds ratio (95% confidence interval, CI)=3.6 (1.5-9.0), P=0.006]. NNRTI-based regimens were intended for 10 patients with resistance to NNRTIs; these patients were prescribed PI-based regimens after genotypic testing. CONCLUSIONS: Transmitted ARV drug resistance was detected in 12.1% of treatment-naive patients, with resistance to NNRTIs the most common. Resistance-testing results played a partial role in first-line treatment selection. However, resistance to NNRTIs pre-empted NNRTI use. PMID- 18400076 TI - Standard-dose efavirenz vs. standard-dose nevirapine in antiretroviral regimens among HIV-1 and tuberculosis co-infected patients who received rifampicin. AB - BACKGROUND: There is limited comparative data between efavirenz (EFV) 600 mg/day and nevirapine (NVP) 400 mg/day-based antiretroviral therapy (ART) among HIV-1 patients with tuberculosis (TB) and receiving rifampicin. METHODS: A retrospective cohort study was conducted in all ART-naive patients who were receiving rifampicin between January 2002 and December 2005. RESULTS: Of 188 patients, 77 and 111 patients were initiated on EFV-based ART (EFV group) and NVP based ART (NVP group), respectively. Overall, median [interquartile range (IQR)] CD4 count was 36 (15-77) cells/microL and median (IQR) viral load was 5.6 (5.2 5.9) HIV-1 RNA log copies/mL. At 48 weeks, 77.9% (60/77) in the EFV group and 67.6% (75/111) in the NVP group achieved HIV-1 RNA <50 copies/mL (P=0.140, odds ratio=0.590, 95% confidence interval=0.302-1.153). At 24 and 48 weeks, respective median CD4 counts were 174 and 254 cells/muL in the EFV group and 156 and 218 cells/microL in the NVP group (P>0.05). By binary logistic regression, treatment group was not associated with HIV-1 RNA <50 copies/mL (P>0.05). No patient in the EFV group and eight (7.2%) patients in the NVP group discontinued ART because of adverse reactions (P=0.084). CONCLUSIONS: For HIV-TB co-infected patients who receive rifampicin, efficacy of 600 mg EFV-based and 400 mg NVP-based ART may be similar, although adverse events tend to be higher in NVP-based ART. PMID- 18400077 TI - Impact of hepatitis B virus co-infection on response to highly active antiretroviral treatment and outcome in HIV-infected individuals: a nationwide cohort study. AB - BACKGROUND: The impact of chronic hepatitis B virus (HBV) infection on viral suppression, immune recovery and mortality in HIV-1 infected patients on highly active antiretroviral treatment (HAART) is a matter of debate. The impact of HBeAg status is unknown. METHODS: This prospective cohort study included all adult Danish HIV-1 infected patients who started HAART between 1 January 1995 and 1 December 2006 (3180 patients). Patients were classified as chronic HBV-infected (6%), HBV-negative (87%) or HBV-unknown (7%). HBV-positive patients were divided into HBeAg-positive or -negative (3.0 vs. 2.6%). Study endpoints were viral load, CD4 cell count and mortality. RESULTS: HBV co-infection had no impact on response to HAART regarding viral suppression or immune recovery. HBV co-infection was associated with several outcomes: overall mortality [mortality rate ratio (MRR) 1.5; 95% confidence interval (CI) 1.1-2.1], liver-related mortality (MRR 4.0; 95% CI 1.6-9.9) and AIDS-related deaths (MRR 1.7; 95% CI 1.0-3.0). The presence of HBeAg did not influence patients' response to HAART. CONCLUSIONS: In HIV patients, chronic HBV infection has no impact on response to HAART concerning viral load and increase in CD4 cell count. However, co-infected patients have an increased mortality compared to HIV-monoinfected patients. PMID- 18400078 TI - Immunological profiles of immune restoration disease presenting as mycobacterial lymphadenitis and cryptococcal meningitis. AB - OBJECTIVES: A proportion of HIV patients beginning antiretroviral therapy (ART) develop immune restoration disease (IRD). Immunological characteristics of IRD were investigated in a cohort of HIV patients beginning therapy in Kuala Lumpur, Malaysia. METHODS: Peripheral blood mononuclear cells were collected at weeks 0, 6, 12, 24 and 48 of ART from five patients experiencing IRD [two with cryptococcal and three with Mycobacterium tuberculosis (Mtb) disease], eight non IRD controls who had begun ART with CD4 T-cell counts of <100 cells/microL and 17 healthy controls. Leukocytes producing interferon-gamma (IFNgamma) were quantified by enzyme-linked immunospot assay after stimulation with purified protein derivative (PPD), early secretory antigenic target-6 (ESAT-6), Cryptococcus neoformans or Cytomegalovirus antigens. Plasma immunoglobulin (IgG) antibodies reactive with these antigens were assessed by enzyme-linked immunosorbent assay. Proportions of activated (HLA-DR(hi)) and regulatory (CD25 CD127(lo) and CTLA-4(+)) CD4 T-cells were quantified by flow cytometry. RESULTS: Plasma HIV RNA declined and CD4 T-cell counts rose within 8-27 weeks on ART. Mtb IRD patients displayed elevated IFNgamma responses and/or plasma IgG to PPD, but none responded to ESAT-6. Cryptococcal IRD occurred in patients with low baseline CD4 T-cell counts and involved clear IFNgamma and antibody responses to cryptococcal antigen. Proportions of activated and regulatory CD4 T-cells declined on ART, but remained higher in patients than in healthy controls. At the time of IRD, proportions of activated CD4 T-cells and regulatory CD4 T-cells were generally elevated relative to other patients. CONCLUSIONS: Cryptococcal and Mtb IRD generally coincide with peaks in the proportion of activated T-cells, pathogen-specific IFNgamma responses and reactive plasma IgG. IRD does not reflect a paucity of regulatory CD4 T-cells. PMID- 18400074 TI - Loss to follow-up in an international, multicentre observational study. AB - OBJECTIVE: The aim of this work was to assess loss to follow-up (LTFU) in EuroSIDA, an international multicentre observational cohort study. METHODS: LTFU was defined as no follow-up visit, CD4 cell count measurement or viral load measurement after 1 January 2006. Poisson regression was used to describe factors related to LTFU. RESULTS: The incidence of LTFU in 12 304 patients was 3.72 per 100 person-years of follow-up [95% confidence interval (CI) 3.58-3.86; 2712 LTFU] and varied among countries from 0.67 to 13.35. After adjustment, older patients, those with higher CD4 cell counts, and those who had started combination antiretroviral therapy all had lower incidences of LTFU, while injecting drug users had a higher incidence of LTFU. Compared with patients from Southern Europe and Argentina, patients from Eastern Europe had over a twofold increased incidence of LTFU after adjustment (incidence rate ratio 2.16; 95% CI 1.84-2.53; P<0.0001). A total of 2743 patients had a period of >1 year with no CD4 cell count or viral load measured during the year; 743 (27.1%) subsequently returned to follow-up. CONCLUSIONS: Some patients thought to be LTFU may have died, and efforts should be made to ascertain vital status wherever possible. A significant proportion of patients who have a year with no follow-up visit, CD4 cell count measurement or viral load measurement subsequently return to follow-up. PMID- 18400079 TI - Antiretroviral drug resistance among antiretroviral-naive persons with recent HIV infection in Thailand. AB - OBJECTIVES: To evaluate the prevalence and patterns of antiretroviral (ARV) drug resistance (ARV-DR) among ARV drug-naive, recently infected persons with HIV in the 4-year interval (2003-2006) after the inception of the National Access to ARV Programme for People who have AIDS in Thailand. METHODS: Cross-sectional study of patients with recent HIV infection for HIV risks, ARV-DR risks and baseline ARV DR. RESULTS: Seven of the 305 patients (2%) had baseline ARV-DR. Via contract tracing, all seven patients with transmitted ARV-DR identified sexual partners with prior ARV treatment failure and had documented low (<75%) ARV adherence. Annual ARV-DR increased from 0 to 5.2% (P=0.06) between 2003 and 2006. CONCLUSIONS: Report of sexual partners with potential HIV and ARV drug exposures can prompt baseline ARV-DR testing of at-risk individuals, while behavioural interventions for adherence and safer sex are refined to minimize the emergence of resistance to generic, fixed-dose combination stavudine, lamivudine and nevirapine (GPO-VIR) therapy. PMID- 18400080 TI - Has carbohydrate-restriction been forgotten as a treatment for diabetes mellitus? A perspective on the ACCORD study design. AB - Prior to the discovery of medical treatment for diabetes, carbohydrate restriction was the predominant treatment recommendation to treat diabetes mellitus. In this commentary we argue that carbohydrate-restriction should be reincorporated into contemporary treatment studies for diabetes mellitus. PMID- 18400081 TI - Genomic context analysis in Archaea suggests previously unrecognized links between DNA replication and translation. AB - BACKGROUND: Comparative analysis of genomes is valuable to explore evolution of genomes, deduce gene functions, or predict functional linking between proteins. Here, we have systematically analyzed the genomic environment of all known DNA replication genes in 27 archaeal genomes to infer new connections for DNA replication proteins from conserved genomic associations. RESULTS: Two distinct sets of DNA replication genes frequently co-localize in archaeal genomes: the first includes the genes for PCNA, the small subunit of the DNA primase (PriS), and Gins15; the second comprises the genes for MCM and Gins23. Other genomic associations of genes encoding proteins involved in informational processes that may be functionally relevant at the cellular level have also been noted; in particular, the association between the genes for PCNA, transcription factor S, and NudF. Surprisingly, a conserved cluster of genes coding for proteins involved in translation or ribosome biogenesis (S27E, L44E, aIF-2 alpha, Nop10) is almost systematically contiguous to the group of genes coding for PCNA, PriS, and Gins15. The functional relevance of this cluster encoding proteins conserved in Archaea and Eukarya is strongly supported by statistical analysis. Interestingly, the gene encoding the S27E protein, also known as metallopanstimulin 1 (MPS-1) in human, is overexpressed in multiple cancer cell lines. CONCLUSION: Our genome context analysis suggests specific functional interactions for proteins involved in DNA replication between each other or with proteins involved in DNA repair or transcription. Furthermore, it suggests a previously unrecognized regulatory network coupling DNA replication and translation in Archaea that may also exist in Eukarya. PMID- 18400082 TI - annot8r: GO, EC and KEGG annotation of EST datasets. AB - BACKGROUND: The expressed sequence tag (EST) methodology is an attractive option for the generation of sequence data for species for which no completely sequenced genome is available. The annotation and comparative analysis of such datasets poses a formidable challenge for research groups that do not have the bioinformatics infrastructure of major genome sequencing centres. Therefore, there is a need for user-friendly tools to facilitate the annotation of non-model species EST datasets with well-defined ontologies that enable meaningful cross species comparisons. To address this, we have developed annot8r, a platform for the rapid annotation of EST datasets with GO-terms, EC-numbers and KEGG-pathways. RESULTS: annot8r automatically downloads all files relevant for the annotation process and generates a reference database that stores UniProt entries, their associated Gene Ontology (GO), Enzyme Commission (EC) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) annotation and additional relevant data. For each of GO, EC and KEGG, annot8r extracts a specific sequence subset from the UniProt dataset based on the information stored in the reference database. These three subsets are then formatted for BLAST searches. The user provides the protein or nucleotide sequences to be annotated and annot8r runs BLAST searches against these three subsets. The BLAST results are parsed and the corresponding annotations retrieved from the reference database. The annotations are saved both as flat files and also in a relational postgreSQL results database to facilitate more advanced searches within the results. annot8r is integrated with the PartiGene suite of EST analysis tools. CONCLUSION: annot8r is a tool that assigns GO, EC and KEGG annotations for data sets resulting from EST sequencing projects both rapidly and efficiently. The benefits of an underlying relational database, flexibility and the ease of use of the program make it ideally suited for non model species EST-sequencing projects. PMID- 18400083 TI - Developmental genetics of the C. elegans pharyngeal neurons NSML and NSMR. AB - BACKGROUND: We are interested in understanding how the twenty neurons of the C. elegans pharynx develop in an intricate yet reproducible way within the narrow confines of the embryonic pharyngeal primordium. To complement an earlier study of the pharyngeal M2 motorneurons, we have now examined the effect of almost forty mutations on the morphology of a bilateral pair of pharyngeal neurosecretory-motor neurons, the NSMs. RESULTS: A careful description of the NSM morphology led to the discovery of a third, hitherto unreported process originating from the NSM cell body and that is likely to play a proprioceptive function. We found that the three NSM processes are differently sensitive to mutations. The major dorsal branch was most sensitive to mutations that affect growth cone guidance and function (e.g. unc-6, unc-34, unc-73), while the major sub-ventral branch was more sensitive to mutations that affect components of the extracellular matrix (e.g. sdn-1). Of the tested mutations, only unc-101, which affects an adaptin, caused the loss of the newly described thin minor process. The major processes developed synaptic branches post-embryonically, and these exhibited activity-dependent plasticity. CONCLUSION: By studying the effects of nearly forty different mutations we have learned that the different NSM processes require different genes for their proper guidance and use both growth cone dependent and growth cone independent mechanisms for establishing their proper trajectories. The two major NSM processes develop in a growth cone dependent manner, although the sub-ventral process relies more on substrate adhesion. The minor process also uses growth cones but uniquely develops using a mechanism that depends on the clathrin adaptor molecule UNC-101. Together with the guidance of the M2 neuron, this is the second case of a pharyngeal neuron establishing one of its processes using an unexpected mechanism. PMID- 18400084 TI - Psychometric characteristics of the Spanish version of instruments to measure neck pain disability. AB - BACKGROUND: The NDI, COM and NPQ are evaluation instruments for disability due to NP. There was no Spanish version of NDI or COM for which psychometric characteristics were known. The objectives of this study were to translate and culturally adapt the Spanish version of the Neck Disability Index Questionnaire (NDI), and the Core Outcome Measure (COM), to validate its use in Spanish speaking patients with non-specific neck pain (NP), and to compare their psychometric characteristics with those of the Spanish version of the Northwick Pain Questionnaire (NPQ). METHODS: Translation/re-translation of the English versions of the NDI and the COM was done blindly and independently by a multidisciplinary team. The study was done in 9 primary care Centers and 12 specialty services from 9 regions in Spain, with 221 acute, subacute and chronic patients who visited their physician for NP: 54 in the pilot phase and 167 in the validation phase. Neck pain (VAS), referred pain (VAS), disability (NDI, COM and NPQ), catastrophizing (CSQ) and quality of life (SF-12) were measured on their first visit and 14 days later. Patients' self-assessment was used as the external criterion for pain and disability. In the pilot phase, patients' understanding of each item in the NDI and COM was assessed, and on day 1 test-retest reliability was estimated by giving a second NDI and COM in which the name of the questionnaires and the order of the items had been changed. RESULTS: Comprehensibility of NDI and COM were good. Minutes needed to fill out the questionnaires [median, (P25, P75)]: NDI. 4 (2.2, 10.0), COM: 2.1 (1.0, 4.9). Reliability: [ICC, (95%CI)]: NDI: 0.88 (0.80, 0.93). COM: 0.85 (0.75,0.91). Sensitivity to change: Effect size for patients having worsened, not changed and improved between days 1 and 15, according to the external criterion for disability: NDI: -0.24, 0.15, 0.66; NPQ: -0.14, 0.06, 0.67; COM: 0.05, 0.19, 0.92. VALIDITY: Results of NDI, NPQ and COM were consistent with the external criterion for disability, whereas only those from NDI were consistent with the one for pain. Correlations with VAS, CSQ and SF-12 were similar for NDI and NPQ (absolute values between 0.36 and 0.50 on day 1, between 0.38 and 0.70 on day 15), and slightly lower for COM (between 0.36 and 0.48 on day 1, and between 0.33 and 0.61 on day 15). Correlation between NDI and NPQ: r = 0.84 on day 1, r = 0.91 on day 15. Correlation between COM and NPQ: r = 0.63 on day 1, r = 0.71 on day 15. CONCLUSION: Although most psychometric characteristics of NDI, NPQ and COM are similar, those from the latter one are worse and its use may lead to patients' evolution seeming more positive than it actually is. NDI seems to be the best instrument for measuring NP-related disability, since its results are the most consistent with patient's assessment of their own clinical status and evolution. It takes two more minutes to answer the NDI than to answer the COM, but it can be reliably filled out by the patient without assistance. TRIAL REGISTRATION: Clinical Trials Register NCT00349544. PMID- 18400085 TI - Chronic disease mortality associated with infectious agents: a comparative cohort study of migrants from the Former Soviet Union in Israel and Germany. AB - BACKGROUND: Prevalence of infectious diseases in migrant populations has been addressed in numerous studies. However, information is sparse on their mortality due to chronic diseases that are aetiologically associated with an infectious agent. This study investigates mortality related to infectious diseases with a specific focus on cancers of possibly infectious origin in voluntary migrants from the Former Soviet Union residing in Israel and in Germany. METHODS: Both groups of migrants arrived from the Former Soviet Union in their destination countries between 1990 and 2001. Population-based data on migrants in Israel were obtained from the Israel Central Bureau of Statistics. Data for migrants in Germany were obtained from a representative sample of all migrants from the Former Soviet Union in Germany. Cause of death information was available until 2003 for the Israeli cohort and until 2005 for the German cohort. Standardized mortality ratios were calculated relative to the destination country for selected causes of death for which infectious agents may be causally involved. Multivariate Poisson regression was applied to assess differences in mortality by length of residence in the host country. RESULTS: Both in Israel and in Germany these migrants have lower overall mortality than the population in their destination countries. However, they have significantly elevated mortality from viral hepatitis and from stomach and liver cancer when compared to the destination populations. Regression analysis shows that in Israel stomach cancer mortality is significantly higher among migrants at shorter durations of residence when compared to durations of more than nine years. CONCLUSION: Higher mortality from cancers associated with infection and from viral hepatitis among migrants from the Former Soviet Union might result from higher prevalence of infections which were acquired in earlier years of life. The results highlight new challenges posed by diseases of infectious origin in migrants and call attention to the link between communicable and non-communicable diseases. PMID- 18400086 TI - Life style related to blood pressure and body weight in adolescence: cross sectional data from the Young-HUNT study, Norway. AB - BACKGROUND: The associations between physical activity, unhealthy dietary habits and cigarette smoking and blood pressure, overweight and obesity are well established in adulthood. This is not the case for similar associations in adolescence. Thus, the purpose of this study is to examine how physical activity, smoking status and dietary habits were related to overweight, obesity and blood pressure in a population of Norwegian adolescents. METHODS: Weight, height, systolic (SBP) and diastolic blood pressure (DBP) were measured, and body mass index (BMI) was calculated among 8408 adolescents who participated in a population based study in 1995-97 in the county of Nord-Trondelag. Internationally accepted cut-off values were used to determine if the adolescents were overweight or obese. The adolescents also completed a detailed questionnaire including dietary habits, physical activity and smoking habits. We calculated adjusted mean blood pressures and odds ratios for being overweight or obese for different exposure categories of life style variables. RESULTS: Low levels of physically activity were associated with increased odds of being overweight (odds ratio (OR), 1.4; 95% confidence interval (CI), 1.1-1.8 in girls and OR, 2.0; 95% CI, 1.6-2.5 in boys) or obese (girls: OR, 3.1; 95% CI, 1.6-6.0; boys: OR, 3.7; 95% CI, 2.1-6.4). In addition, the least physically active girls had a 1.5 mmHg higher mean DBP compared with the most active (p-trend <0.001), and among boys this difference was 1.0 mmHg (p-trend < 0.001). Smokers were more likely to be obese (OR, 1.6; 95% CI, 1.1-2.5 in girls and 1.4; 95% CI, 0.9-2.1 in boys) compared with non-smokers. Smokers also had lower mean SBP than non-smokers; however, this finding was restricted to smokers with the lowest smoking exposure. Associations between dietary habits and weight status largely disappeared after adjusting for weight losing behaviour. CONCLUSION: In this population of adolescents low levels of physical activity were associated with higher mean DBP and higher odds of overweight or obesity. Smoking was also associated with higher odds of overweight and obesity. The paradoxical associations between healthy dietary habits and overweight and obesity are most likely an effect of reverse causality. PMID- 18400087 TI - Molecular and functional characterization of a Rho GDP dissociation inhibitor in the filamentous fungus Tuber borchii. AB - BACKGROUND: Small GTPases of the Rho family function as tightly regulated molecular switches that govern important cellular functions in eukaryotes. Several families of regulatory proteins control their activation cycle and subcellular localization. Members of the guanine nucleotide dissociation inhibitor (GDI) family sequester Rho GTPases from the plasma membrane and keep them in an inactive form. RESULTS: We report on the characterization the RhoGDI homolog of Tuber borchii Vittad., an ascomycetous ectomycorrhizal fungus. The Tbgdi gene is present in two copies in the T. borchii genome. The predicted amino acid sequence shows high similarity to other known RhoGDIs. Real time PCR analyses revealed an increased expression of Tbgdi during the phase preparative to the symbiosis instauration, in particular after stimulation with root exudates extracts, that correlates with expression of Tbcdc42. In a translocation assay TbRhoGDI was able to solubilize TbCdc42 from membranes. Surprisingly, TbRhoGDI appeared not to interact with S. cerevisiae Cdc42, precluding the use of yeast as a surrogate model for functional studies. To study the role of TbRhoGDI we performed complementation experiments using a RhoGDI null strain of Dictyostelium discoideum, a model organism where the roles of Rho signaling pathways are well established. For comparison, complementation with mammalian RhoGDI1 and LyGDI was also studied in the null strain. Although interacting with Rac1 isoforms, TbRhoGDI was not able to revert the defects of the D. discoideum RhoGDI null strain, but displayed an additional negative effect on the cAMP-stimulated actin polymerization response. CONCLUSION: T. borchii expresses a functional RhoGDI homolog that appears as an important modulator of cytoskeleton reorganization during polarized apical growth that antecedes symbiosis instauration. The specificity of TbRhoGDI actions was underscored by its inability to elicit a growth defect in S. cerevisiae or to compensate the loss of a D. discoideum RhoGDI. Knowledge of the cell signaling at the basis of cytoskeleton reorganization of ectomycorrhizal fungi is essential for improvements in the production of mycorrhized plant seedlings used in timberland extension programs and fruit body production. PMID- 18400088 TI - Mapping of mitral regurgitant defects by cardiovascular magnetic resonance in moderate or severe mitral regurgitation secondary to mitral valve prolapse. AB - PURPOSE: In mitral valve prolapse, determining whether the valve is suitable for surgical repair depends on the location and mechanism of regurgitation. We assessed whether cardiovascular magnetic resonance (CMR) could accurately identify prolapsing or flail mitral valve leaflets and regurgitant jet direction in patients with known moderate or severe mitral regurgitation. METHODS: CMR of the mitral valve was compared with trans-thoracic echocardiography (TTE) in 27 patients with chronic moderate to severe mitral regurgitation due to mitral valve prolapse. Contiguous long-axis high temporal resolution CMR cines perpendicular to the valve commissures were obtained across the mitral valve from the medial to lateral annulus. This technique allowed systematic valve inspection and mapping of leaflet prolapse using a 6 segment model. CMR mapping was compared with trans oesophageal echocardiography (TOE) or surgical inspection in 10 patients. RESULTS: CMR and TTE agreed on the presence/absence of leaflet abnormality in 53 of 54 (98%) leaflets. Prolapse or flail was seen in 36 of 54 mitral valve leaflets examined on TTE. CMR and TTE agreed on the discrimination of prolapse from flail in 33 of 36 (92%) leaflets and on the predominant regurgitant jet direction in 26 of the 27 (96%) patients. In the 10 patients with TOE or surgical operative findings available, CMR correctly classified presence/absence of segmental abnormality in 49 of 60 (82%) leaflet segments. CONCLUSION: Systematic mitral valve assessment using a simple protocol is feasible and could easily be incorporated into CMR studies in patients with mitral regurgitation due to mitral valve prolapse. PMID- 18400089 TI - Quantitative characterization of myocardial infarction by cardiovascular magnetic resonance predicts future cardiovascular events in patients with ischemic cardiomyopathy. AB - BACKGROUND: Cardiovascular magnetic resonance (CMR) can provide quantitative data of the myocardial tissue utilizing high spatial and temporal resolution along with exquisite tissue contrast. Previous studies have correlated myocardial scar tissue with the occurrence of ventricular arrhythmia. This study was conducted to evaluate whether characterization of myocardial infarction by CMR can predict cardiovascular events in patients with ischemic cardiomyopathy (ICM). RESULTS: We consecutively studied 86 patients with ICM (LVEF < 50%, mean LVEF: 26 +/- 12%) with CMR before revascularization or medication therapy +/- implantable cardiac defibrillator, determined the amount of myocardial scar, and followed for development of cardiovascular events. Thirty-three patients (38%) had cardiovascular events (mean follow-up: 20 +/- 16 months). Patients who developed cardiovascular events had larger scar volume and scar percentage of the myocardium than those who did not develop cardiovascular events (16.8 +/- 12.4 cm3 vs. 11.7 +/- 12.6 cm3, p = 0.023 and 10.2 +/- 6.9% vs. 7.2 +/- 6.7%, p = 0.037, respectively). There were no significant differences in LVEDV, LVESV and LVEF between the patients with and without cardiovascular events (231 +/- 76 ml vs. 230 +/- 88 ml; 180 +/- 73 ml vs. 175 +/- 90 ml; and 25 +/- 10% vs. 27 +/- 13%, respectively). CONCLUSION: Quantification of the scar volume and scar percentage by CMR is superior to LVEDV, LVESV, and LVEF in prognosticating the future likelihood of the development of cardiovascular events in patients with ICM. PMID- 18400090 TI - Research utilization among children's mental health providers. AB - BACKGROUND: Children with emotional and behavioural disorders should be able to count on receiving care that meets their needs and is based on the best scientific evidence available, however, many do not receive these services. Implementation of evidence-based practice (EBP) relies, in part, on the research utilization practices of mental health care providers. This study reports on a survey of research utilization practices among 80 children's mental health (CMH) service provider organizations in Ontario, Canada. METHODS: A web-based survey was distributed to 80 CMH service provider organizations, to which 51 executive directors and 483 children's mental health practitioners responded. Research utilization was assessed using questions with Likert-type responses based on the Canadian Health Services Research Foundation's Four-A's approach: access, assess, adapt, apply. RESULTS: There was general agreement among executive directors and practitioners regarding the capacity of their organizations to use - access, assess, adapt, and apply - research evidence. Overall, both groups rated their organizations as using research information 'somewhat well.' The low response rate to the practitioner survey should be noted. CONCLUSION: These findings provide a useful benchmark from which changes in reported research utilization in the Ontario CMH sector can be tracked over time, as a function of EBP training and implementation initiatives, for instance. The need to improve access to research evidence should be addressed because it relates to the eventual implementation and uptake of evidence-based practices. Communities of practice are recommended as a strategy that would enable practitioners to build capacity in their adaptation and application of research evidence. PMID- 18400091 TI - Body Dysmorphic Disorder: gender differences and prevalence in a Pakistani medical student population. AB - BACKGROUND: Body dysmorphic disorder (BDD) is a psychiatric disorder characterized by a preoccupation with an imagined or slight defect which causes significant distress or impairment in functioning. Few studies have assessed gender differences in BDD in a non clinical population. Also no study assessed BDD in medical students. This study was designed to determine the point prevalence of BDD in Pakistani medical students and the gender differences in prevalence of BDD, body foci of concern and symptoms of BDD. METHODS: The medical students enrolled in a medical university in Karachi, Pakistan filled out a self report questionnaire which assessed clinical features of BDD. BDD was diagnosed according to the DSM-IV criteria. RESULTS: Out of the 156 students, 57.1% were female. A total of 78.8% of the students reported dissatisfaction with some aspect of their appearance and 5.8% met the DSM-IV criteria for BDD. The male to female ratio for BDD was 1.7. Regarding gender differences in body foci of concern, the top three reported foci of concern in male students were head hair (34.3%), being fat (32.8%), skin (14.9%) and nose(14.9%), whereas in females they were being fat (40.4%), skin (24.7%) and teeth (18%). Females were significantly more concerned about being fat (p = 0.005). Male students were significantly more concerned about being thin (p = 0.01) and about head hair (p = 0.012). CONCLUSION: BDD is fairly common in our medical student population, with a higher prevalence in males. Important gender differences in BDD symptomatology and reported body foci of concern were identified which reflected the influence of media on body image perception. The impact of cultural factors on the prevalence as well as gender differences in BDD symptomatology was also established. PMID- 18400092 TI - Does pharmaceutical advertising affect journal publication about dietary supplements? AB - BACKGROUND: Advertising affects consumer and prescriber behaviors. The relationship between pharmaceutical advertising and journals' publication of articles regarding dietary supplements (DS) is unknown. METHODS: We reviewed one year of the issues of 11 major medical journals for advertising and content about DS. Advertising was categorized as pharmaceutical versus other. Articles about DS were included if they discussed vitamins, minerals, herbs or similar products. Articles were classified as major (e.g., clinical trials, cohort studies, editorials and reviews) or other (e.g., case reports, letters, news, and others). Articles' conclusions regarding safety and effectiveness were coded as negative (unsafe or ineffective) or other (safe, effective, unstated, unclear or mixed). RESULTS: Journals' total pages per issue ranged from 56 to 217 while advertising pages ranged from 4 to 88; pharmaceutical advertisements (pharmads) accounted for 1.5% to 76% of ad pages. Journals with the most pharmads published significantly fewer major articles about DS per issue than journals with the fewest pharmads (P < 0.01). Journals with the most pharmads published no clinical trials or cohort studies about DS. The percentage of major articles concluding that DS were unsafe was 4% in journals with fewest and 67% among those with the most pharmads (P = 0.02). The percentage of articles concluding that DS were ineffective was 50% higher among journals with more than among those with fewer pharmads (P = 0.4). CONCLUSION: These data are consistent with the hypothesis that increased pharmaceutical advertising is associated with publishing fewer articles about DS and publishing more articles with conclusions that DS are unsafe. Additional research is needed to test alternative hypotheses for these findings in a larger sample of more diverse journals. PMID- 18400093 TI - GPR50 is the mammalian ortholog of Mel1c: evidence of rapid evolution in mammals. AB - BACKGROUND: The melatonin receptor subfamily contains three members Mel1a, Mel1b and Mel1c, found in all vertebrates except for Mel1c which is found only in fish, Xenopus species and the chicken. Another receptor, the melatonin related receptor known as GPR50, found exclusively in mammals and later identified as a member of the melatonin receptor subfamily because of its identity to the three melatonin receptors despite its absence of affinity for melatonin. The aim of this study was to describe the evolutionary relationships between GPR50 and the three other members of the melatonin receptor subfamily. RESULTS: Using an in silico approach, we demonstrated that GPR50 is the ortholog of the high affinity Mel1c receptor. It was necessary to also study the synteny of this gene to reach this conclusion because classical mathematical models that estimate orthology and build phylogenetic trees were not sufficient. The receptor has been deeply remodelled through evolution by the mutation of numerous amino acids and by the addition of a long C-terminal tail. These alterations have modified its affinity for melatonin and probably affected its interactions with the other two known melatonin receptors MT1 and MT2 that are encoded by Mel1a and Mel1b genes respectively. Evolutionary studies provided evidence that the GPR50 group evolved under different selective pressure as compared to the orthologous groups Me11 a, b, and c. CONCLUSION: This study demonstrated that there are only three members in the melatonin receptor subfamily with one of them (Me11c) undergoing rapid evolution from fishes and birds to mammals. Further studies are necessary to investigate the physiological roles of this receptor. PMID- 18400094 TI - Altered regulation of Prox1-gene-expression in liver tumors. AB - BACKGROUND: Prospero-related homeobox 1 (Prox1) transcription factor was described as a tumor-suppressor gene in liver tumors. In contrast, Prox1 knock out in murine embryos drastically reduces proliferation of hepatoblasts. METHODS: We have studied the expression of Prox1 in normal liver, liver cirrhosis and peritumoral liver samples in comparison to hepatocellular (HCC) and cholangiocellular carcinoma (CCC) at mRNA, protein and functional levels. RESULTS: Prox1 was found in hepatocytes of normal liver, while normal bile duct epithelial cells were negative. However, Prox1+ cells, which co-expressed biliary epithelial makers and showed ductular morphology, could be detected within fibrotic septa of cirrhotic livers, and in both HCC and CCC. Two Prox1 mRNA isoforms (2.9 kb and 7.9 kb) were identified with a prevalence of the longer isoform in several HCC samples and the shorter in most CCC samples. Evidence was provided that Myc-associated zinc finger protein (MAZ) might significantly contribute to the gene expression of Prox1 in HCC, while neo-expression of Prox1 in CCC remains to be resolved. A point mutation in the prospero domain of Prox1 was found in one HCC sample. CONCLUSION: Our study shows dysregulation of Prox1 in liver cirrhosis, HCC and CCC, such as neo-expression in cells with biliary epithelial phenotype in liver cirrhosis, and in CCC. Altered Prox1 mRNA expression is partly regulated by MAZ, and mutation of the prospero domain in HCC indicates an involvement for Prox1 during tumor progression. PMID- 18400095 TI - Postpartum ovarian vein thrombosis after cesarean delivery: a case report. AB - INTRODUCTION: Postpartum ovarian vein thrombosis is an uncommon complication; incidence varies between 0.002% and 0.05%. It most often occurs during the 2-15 days following delivery. CASE PRESENTATION: A 22-year-old pregnant woman at term presented to hospital with uterine contractions, abdominal pain, nausea and vomiting. After delivery an ovarian vein thrombosis was diagnosed. CONCLUSION: Low-molecular weight heparin with broad-spectrum antibiotics are the accepted therapy in non-complicated cases of postpartum ovarian vein thrombosis. PMID- 18400096 TI - Case-mix & patients' reports of outcome in Independent Sector Treatment Centres: Comparison with NHS providers. AB - BACKGROUND: There has been considerable concern expressed about the outcomes achieved in Independent Sector Treatment Centres (ISTCs) introduced in England since 2003. Our aim was to compare the case-mix and patients' reported outcomes of surgery in ISTCs and in NHS providers. METHODS: Prospective cohort study of 769 patients treated in six ISTCs and 1895 treated in 20 NHS providers (acute hospitals and treatment centres) in England during 2006-07. Participants underwent one of three day surgery procedures (inguinal hernia repair, varicose vein surgery, cataract extraction) or hip or knee replacement. Change in patient reported health status and health related quality of life (measured using a disease-specific and a generic (EQ-5D) instrument) was assessed either 3-months (day surgery) or 6-months (hip/knee) after surgery. In addition patient-reported post-operative complications and an overall assessment of success of surgery were collected. Outcome measures were adjusted (using multivariable regression) for patient characteristics (disease severity, duration of symptoms, age, sex, socioeconomic status, general health, previous similar surgery, comorbidity). RESULTS: Post-operative response rates varied by procedure (73%-88%) and were similar for those treated in ISTCs and NHS facilities. Patients treated in ISTCs were healthier, were less likely to have any comorbidity and, for those undergoing cataract surgery or joint replacement, their primary condition was less severe. Those undergoing hernia repair or joint replacement were less likely to have had similar surgery before. When adjustment was made for pre-operative characteristics, patients undergoing cataract surgery or hip replacement in ISTCs achieved a slightly greater improvement in functional status and quality of life than those treated in NHS facilities, while the opposite was true of patients undergoing hernia repair. No significant differences were found for the two other procedures. Patients treated in ISTCs were less likely to report post-operative problems than those treated in NHS facilities for cataract surgery (Adjusted Odds Ratio 0.35; 95% CI 0.17-0.70), hernia repair (0.42; 0.28-0.63) and knee replacement (0.44; 0.28-0.69). Most patients described the result of their operation as excellent, very good or good, regardless of where they were treated. CONCLUSION: The case-mix of patients treated in ISTCs differs from that in NHS providers, in line with the intention of the contracts. Caution is needed in interpreting the observation that patients treated in ISTCs reported slightly better outcomes as very few ISTCs participated, case-mix adjustment might have been insufficient, and patients' reports might have been biased as they were more likely to be satisfied with the way they were treated. PMID- 18400097 TI - Identification of a novel KCNQ1 mutation associated with both Jervell and Lange Nielsen and Romano-Ward forms of long QT syndrome in a Chinese family. AB - BACKGROUND: Long QT syndrome (LQTS) is a cardiac disorder characterized by prolonged QT intervals on electrocardiograms (ECG), ventricular arrhythmias, and sudden death. Clinically, two inherited forms of LQTS have been defined: autosomal dominant LQTS or Romano-Ward syndrome (RWS) not associated with deafness and autosomal recessive LQTS or Jervell and Lange-Nielsen syndrome (JLNS) associated with deafness. METHODS: A Chinese family with both RWS and JLNS was identified. Family members were diagnosed based on the presence of a prolonged QT interval as seen on a 12-lead ECG and a medical history of syncope, palpitation, and deafness. Mutational studies in the KCNQ1 potassium channel gene were performed using direct DNA sequence analysis and restriction length polymorphism analysis. RESULTS: The proband in the Chinese family and her brother had previously been diagnosed with JLNS, and two other members were affected with RWS. The proband was also affected with atrial fibrillation. A single nucleotide substitution of C to T at nucleotide 965 of KCNQ1 was identified, and the mutation resulted in the substitution of a threonine residue at codon 322 by a methionine residue (T322M). The novel heterozygous T322M mutation was identified in two patients with RWS, one member with borderline QTc, and two normal family members. The two JLNS patients in the family carried the homozygous T322M mutation. The T322M mutation was not found in 200 Chinese normal controls. CONCLUSION: Our results suggest that T322M is a novel mutation that caused RWS with high intrafamilial variability in the heterozygous carriers and typical JLNS in the homozygous carriers within this Chinese family. The T322M mutation is the first mutation identified for JLNS in the Chinese population. PMID- 18400098 TI - Review for the generalist: evaluation of pediatric foot and ankle pain. AB - Foot and ankle pain is common in children and adolescents. Problems are usually related to skeletal maturity and are fairly specific to the age of the child. Evaluation and management is challenging and requires a thorough history and physical exam, and understanding of the pediatric skeleton. This article will review common causes of foot and ankle pain in the pediatric population. PMID- 18400099 TI - PSAIA - protein structure and interaction analyzer. AB - BACKGROUND: PSAIA (Protein Structure and Interaction Analyzer) was developed to compute geometric parameters for large sets of protein structures in order to predict and investigate protein-protein interaction sites. RESULTS: In addition to most relevant established algorithms, PSAIA offers a new method PIADA (Protein Interaction Atom Distance Algorithm) for the determination of residue interaction pairs. We found that PIADA produced more satisfactory results than comparable algorithms implemented in PSAIA. Particular advantages of PSAIA include its capacity to combine different methods to detect the locations and types of interactions between residues and its ability, without any further automation steps, to handle large numbers of protein structures and complexes. Generally, the integration of a variety of methods enables PSAIA to offer easier automation of analysis and greater reliability of results.PSAIA can be used either via a graphical user interface or from the command-line. Results are generated in either tabular or XML format. CONCLUSION: In a straightforward fashion and for large sets of protein structures, PSAIA enables the calculation of protein geometric parameters and the determination of location and type for protein protein interaction sites. XML formatted output enables easy conversion of results to various formats suitable for statistic analysis. Results from smaller data sets demonstrated the influence of geometry on protein interaction sites. Comprehensive analysis of properties of large data sets lead to new information useful in the prediction of protein-protein interaction sites. PMID- 18400100 TI - Socioeconomic position and risk of short-term weight gain: prospective study of 14,619 middle-aged men and women. AB - BACKGROUND: The association between socioeconomic position in middle age and risk of subsequent, short-term weight gain is unknown. We therefore assessed this association in a prospective population based cohort study in Norfolk, UK. METHODS: We analysed data on 14,619 middle-aged men and women (aged between 40-75 at baseline) with repeated objective measures of weight and height at baseline (1993-1997) and follow up (1998-2000). RESULTS: During follow up 5,064 people gained more than 2.5 kg. Compared with the highest social class, individuals in the lowest social class had around a 30% greater risk of gaining more than 2.5 kg (OR 1.29; 95% CI 1.11-1.51; p for trend = 0.002). This association remained statistically significant following adjustment for sex, age, baseline BMI, smoking, and follow up time (OR 1.25; CI 1.07-1.46; p for trend <0.001). We also found no material difference between unadjusted models and those including all confounders and potential mediators. CONCLUSION: Individuals of low socioeconomic position are at greatest risk of gaining weight during middle age, which is not explained by classical correlates of socioeconomic position and risk factors for obesity. PMID- 18400102 TI - Gender differences in sex life issues - a population-based study of migraine sufferers. AB - BACKGROUND: Migraine is considered to have a negative influence on sex life. The present study was to analyse the perceptions of importance of and satisfaction with sex life as well as the expression of interest in sex among people having migraines in a prospective follow-up mail survey in 1998 and 2003. METHODS: The random sample was stratified according to gender and age in four age groups (20 24, 30-34, 40-44, and 50-54 years). Altogether 25 898 individuals responded to the baseline and 19 626 to the follow-up questionnaire (75.8% response rate). We examined as to how the perceptions of sex life of those suffering from migraine changed during a 5-year follow-up. Conditional logistic regression was used to analyse the data of the responses on self-reported migraine in the baseline and follow-up surveys (N = 2 977, 79.2% women). Each person with migraine was assigned a gender- and age-matched control in the analysis. RESULTS: All three outcome variables tended to decrease in value. Importance of sex life was higher among men with migraine than among their controls. Among women migraine lessened interest in sex life. CONCLUSION: Our findings suggested that migraine has a different impact on sex life among women from that among men. PMID- 18400101 TI - Exercise alters the immune profile in Tg2576 Alzheimer mice toward a response coincident with improved cognitive performance and decreased amyloid. AB - BACKGROUND: Inflammation is associated with Abeta pathology in Alzheimer's disease (AD) and transgenic AD models. Previously, it has been demonstrated that chronic stimulation of the immune response induces pro-inflammatory cytokines IL 1beta and TNF-alpha which contribute to neurodegeneration. However, recent evidence has shown that inducing the adaptive immune response reduces Abeta pathology and is neuroprotective. Low concentrations of IFN-gamma modulate the adaptive immune response by directing microglia to differentiate to antigen presenting cells. Our objective was to determine if exercise could induce a shift from the immune profile in aged (17-19 months) Tg2576 mice to a response that reduces Abeta pathology. METHODS: TG (n = 29) and WT (n = 27) mice were divided into sedentary (SED) and exercised (RUN) groups. RUN animals were provided an in cage running wheel for 3 weeks. Tissue was harvested and hippocampus and cortex dissected out. Quantitative data was analyzed using 2 x 2 ANOVA and student's t tests. RESULTS: IL-1beta and TNF-alpha were significantly greater in hippocampi from sedentary Tg2576 (TGSED) mice than in wildtype (WTSED) (p = 0.04, p = 0.006). Immune response proteins IFN-gamma and MIP-1alpha are lower in TGSED mice than in WTSED (p = 0.03, p = 0.07). Following three weeks of voluntary wheel running, IL-1beta and TNF-alpha decreased to levels indistinguishable from WT. Concurrently, IFN-gamma and MIP-1alpha increased in TGRUN. Increased CD40 and MHCII, markers of antigen presentation, were observed in TGRUN animals compared to TGSED, as well as CD11c staining in and around plaques and vasculature. Additional vascular reactivity observed in TGRUN is consistent with an alternative activation immune pathway, involving perivascular macrophages. Significant decreases in soluble Abeta40 (p = 0.01) and soluble fibrillar Abeta (p = 0.01) were observed in the exercised transgenic animals. CONCLUSION: Exercise shifts the immune response from innate to an adaptive or alternative response. This shift in immune response coincides with a decrease in Abeta in advanced pathological states. PMID- 18400103 TI - Comparative transcriptome analysis of Arabidopsis thaliana infested by diamond back moth (Plutella xylostella) larvae reveals signatures of stress response, secondary metabolism, and signalling. AB - BACKGROUND: Plants are exposed to attack from a large variety of herbivores. Feeding insects can induce substantial changes of the host plant transcriptome. Arabidopsis thaliana has been established as a relevant system for the discovery of genes associated with response to herbivory, including genes for specialized (i.e. secondary) metabolism as well as genes involved in plant-insect defence signalling. RESULTS: Using a 70-mer oligonucleotide microarray covering 26,090 gene-specific elements, we monitored changes of the Arabidopsis leaf transcriptome in response to feeding by diamond back moth (DBM; Plutella xylostella) larvae. Analysis of samples from a time course of one hour to 24 hours following onset of DBM feeding revealed almost three thousand (2,881) array elements (including 2,671 genes with AGI annotations) that were differentially expressed (>2-fold; p[t-test] < 0.05) of which 1,686 also changed more than twofold in expression between at least two time points of the time course with p(ANOVA) < 0.05. While the majority of these transcripts were up-regulated within 8 h upon onset of insect feeding relative to untreated controls, cluster analysis identified several distinct temporal patterns of transcriptome changes. Many of the DBM-induced genes fall into ontology groups annotated as stress response, secondary metabolism and signalling. Among DBM-induced genes associated with plant signal molecules or phytohormones, genes associated with octadecanoid signalling were clearly overrepresented. We identified a substantial number of differentially expressed genes associated with signal transduction in response to DBM feeding, and we compared there expression profiles with those of previously reported transcriptome responses induced by other insect herbivores, specifically Pieris rapae, Frankliniella occidentalis, Bemisia tabaci,Myzus persicae, and Brevicoryne brassicae. CONCLUSION: Arabidopsis responds to feeding DBM larvae with a drastic reprogramming of the transcriptome, which has considerable overlap with the response induced by other insect herbivores. Based on a meta-analysis of microarray data we identified groups of transcription factors that are either affected by multiple forms of biotic or abiotic stress including DBM feeding or, alternatively, were responsive to DBM herbivory but not to most other forms of stress. PMID- 18400104 TI - Detailed characterization of the mouse embryonic stem cell transcriptome reveals novel genes and intergenic splicing associated with pluripotency. AB - BACKGROUND: Transcriptional control of embryonic stem (ES) cell pluripotency has been a subject of intense study. Transcriptional regulators including Oct4 (Oct3/4 index), Sox2 and Nanog are fundamental for maintaining the undifferentiated state. However, the ES cell transcriptome is not limited to their targets, and exhibits considerable complexity when assayed with microarray, MPSS, cDNA/EST sequencing, and SAGE technologies. To identify novel genes associated with pluripotency, we globally searched for ES transcripts not corresponding to known genes, validated their sequences, determined their expression profiles, and employed RNAi to test their function. RESULTS: Gene Identification Signature (GIS) analysis, a SAGE derivative distinguished by paired 5' and 3' transcript end tags, identified 153 candidate novel transcriptional units (TUs) distinct from known genes in a mouse E14 ES mRNA library. We focused on 16 TUs free of artefacts and mapping discrepancies, five of which were validated by RTPCR product sequencing. Two of the TUs were revealed by annotation to represent novel protein-coding genes: a PRY-domain cluster member and a KRAB-domain zinc finger. The other three TUs represented intergenic splicing events involving adjacent, functionally unrelated protein-coding genes transcribed in the same orientation, with one event potentially encoding a fusion protein containing domains from both component genes (Clk2 and Scamp3). Expression profiling using embryonic samples and adult tissue panels confirmed that three of the TUs were unique to or most highly expressed in ES cells. Expression levels of all five TUs dropped dramatically during three distinct chemically induced differentiation treatments of ES cells in culture. However, siRNA knockdowns of the TUs did not alter mRNA levels of pluripotency or differentiation markers, and did not affect cell morphology. CONCLUSION: Transcriptome libraries retain considerable potential for novel gene discovery despite massive recent cDNA and EST sequencing efforts; cDNA and EST evidence for these ES cell TUs had been limited or absent. RTPCR and full-length sequencing remain essential in resolving the bottleneck between numerous candidate novel transcripts inferred from high-throughput sequencing and the small fraction that can be validated. RNAi results indicate that, despite their strong association with pluripotency, these five transcriptomic novelties may not be required for maintaining it. PMID- 18400105 TI - Very mild disease phenotype of congenic CftrTgH(neoim)Hgu cystic fibrosis mice. AB - BACKGROUND: A major boost to cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)Hgu mouse model with a divergent genetic background (129/Sv, C57BL/6, HsdOla:MF1) two inbred mutant mouse strains CF/1 CftrTgH(neoim)Hgu and CF/3-CftrTgH(neoim)Hgu had been generated using strict brother x sister mating. CF/1-CftrTgH(neoim)Hgu and CF/3-CftrTgH(neoim)Hgu mice were fertile and showed normal growth and lifespan. In this work the CftrTgH(neoim)Hgu insertional mutation was backcrossed from CF/3 CftrTgH(neoim)Hgu onto the inbred backgrounds C57BL/6J and DBA/2J generating congenic animals in order to clarify the differential impact of the Cftr mutation and the genetic background on the disease phenotype of the cystic fibrosis mutant mice. Clinical and electrophysiological features of the two congenic strains were compared with those of CF/1-CftrTgH(neoim)Hgu and CF/3-CftrTgH(neoim)Hgu and wild type controls. RESULTS: Under the standardized housing conditions of the animal facility, the four mouse strains CF/1-CftrTgH(neoim)Hgu, CF/3-CftrTgH(neoim)Hgu, D2.129P2(CF/3)-CftrTgH(neoim)Hgu and B6.129P2(CF/3)-CftrTgH(neoim)Hgu exhibited normal life expectancy. Growth of congenic cystic fibrosis mice was comparable with that of wild type controls. All mice but D2.129P2(CF/3)-CftrTgH(neoim)Hgu females were fertile. Short circuit current measurements revealed characteristic response profiles of the HsdOla:MF1, DBA/2J and C57BL/6J backgrounds in nose, ileum and colon. All cystic fibrosis mouse lines showed the disease-typical hyperresponsiveness to amiloride in the respiratory epithelium. The mean chloride secretory responses to carbachol or forskolin were 15-100% of those of the cognate wild type control animals. CONCLUSION: The amelioration of the clinical features and of the basic defect that had emerged during the generation of CF/3 CftrTgH(neoim)Hgu mice was retained in the congenic mice indicating that the Cftr linkage group or other loci shared between the inbred strains contain(s) the major modifier(s) of attenuation of cystic fibrosis symptoms. PMID- 18400106 TI - Undergraduate technical skills training guided by student tutors--analysis of tutors' attitudes, tutees' acceptance and learning progress in an innovative teaching model. AB - BACKGROUND: Skills labs provide a sheltered learning environment. As close supervision and individual feedback were proven to be important in ensuring effective skills training, we implemented a cross-year peer tutor system in our skills lab of internal medicine that allowed intense training sessions with small learning groups (3-4 students) taught by one student tutor. METHODS: The expectations, experiences and criticisms of peer tutors regarding the tutor system for undergraduate skills lab training were investigated in the context of a focus group. In addition, tutees' acceptance of this learning model and of their student tutors was evaluated by means of a pre/post web-based survey. RESULTS: 14 voluntary senior students were intensely prepared by consultants for their peer tutor activity. 127 students participated in the project, 66.9% of which responded to the web-based survey (23 topics with help of 6-point Likert scale + free comments). Acceptance was very high (5.69 +/- 0.07, mean +/- SEM), and self-confidence ratings increased significantly after the intervention for each of the trained skills (average 1.96 +/- 0.08, all p < 0.002). Tutors received high global ratings (5.50 +/- 0.07) and very positive anonymous individual feedback from participants. 82% of tutees considered the peer teaching model to be sufficient, and a mere 1% expressed the wish for skills training to be provided by faculty staff only. Focus group analyses with tutors revealed 18 different topics, including profit in personal knowledge and personal satisfaction through teaching activities. The ratio of 1:4 tutor/tutees was regarded to be very beneficial for effective feedback, and the personalized online evaluation by tutees to be a strong motivator and helpful for further improvements. The tutors ascribed great importance to the continuous availability of a contact doctor in case of uncertainties. CONCLUSION: This study demonstrates that peer teaching in undergraduate technical clinical skills training is feasible and widely accepted among tutees, provided that the tutors receive sufficient training and supervision. PMID- 18400107 TI - Steroid hormone regulation of EMP2 expression and localization in the endometrium. AB - BACKGROUND: The tetraspan protein epithelial membrane protein-2 (EMP2), which mediates surface display of diverse proteins, is required for endometrial competence in blastocyst implantation, and is uniquely correlated with poor survival from endometrial adenocarcinoma tumors. Because EMP2 is differentially expressed in the various stages of the murine and human estrous cycle, we tested the hypothesis that the steroid hormones progesterone and estrogen influence EMP2 expression and localization. METHODS: Frozen human proliferative and secretory endometrium were collected and analyzed for EMP2 expression using SDS PAGE/Western blot analysis. The response of EMP2 to progesterone and estradiol was determined using a combination of real-time PCR, SDS-PAGE/Western blot analysis, and confocal immunofluorescence in the human endometrial carcinoma cell line RL95-2. To confirm the in vitro results, ovariectomized mice were treated with progesterone or estradiol, and EMP2 expression was analyzed using immunohistochemistry. RESULTS: Within normal human endometrium, EMP2 expression is upregulated in the secretory phase relative to the proliferative phase. To understand the role of steroid hormones on EMP2 expression, we utilized RL95-2 cells, which express both estrogen and progesterone receptors. In RL95-2 cells, both estradiol and progesterone induced EMP2 mRNA expression, but only progesterone induced EMP2 protein expression. To compare steroid hormone regulation of EMP2 between humans and mice, we analyzed EMP2 expression in ovarectomized mice. Similar to results observed in humans, progesterone upregulated endometrial EMP2 expression and induced EMP2 translocation to the plasma membrane. Estradiol did not promote translocation to the cell surface, but moderately induced EMP2 expression in cytoplasmic compartments in vivo. CONCLUSION: These findings suggest that targeting of EMP2 to specific locations under the influence of these steroid hormones may be important for integrating the molecular responses required for implantation competence. PMID- 18400108 TI - Application of Portsmouth modification of physiological and operative severity scoring system for enumeration of morbidity and mortality (P-POSSUM) in pancreatic surgery. AB - BACKGROUND: Pancreatoduodenectomy (PD) is associated with high incidence of morbidity and mortality. We have applied P-POSSUM in predicting the incidence of outcome after PD to identify those who are at the highest risk of developing complications. METHOD: A prospective database of 241 consecutive patients who had PD from January 2002 to September 2005 was retrospectively updated and analysed. P-POSSUM score was calculated for each patient and correlated with observed morbidity and mortality. RESULTS: 30 days mortality was 7.8% and morbidity was 44.8%. Mean physiological score was 16.07 +/- 3.30. Mean operative score was 13.67 +/- 3.42. Mean operative score rose to 20.28 +/- 2.52 for the complex major operation (p < 0.001) with 2 fold increase in morbidity and 3.5 fold increase in mortality. For groups of patients with a physiological score of (less than or equal to) 18, the O:P (observed to Predicted) morbidity ratio was 1.3-1.4 and, with a physiological score of >18, the O:P ratio was nearer to 1. Physiological score and white cell count were significant in a multivariate model. CONCLUSION: P-POSSUM underestimated the mortality rate. While P-POSSUM analysis gave a truer prediction of morbidity, underestimation of morbidity and potential for systematic inaccuracy in prediction of complications at lower risk levels is a significant issue for pancreatic surgery. PMID- 18400110 TI - Elevated maternal lipoprotein (a) and neonatal renal vein thrombosis: a case report. AB - INTRODUCTION: Renal vein thrombosis, although rare in adults, is well recognized in neonates and is one of the most common manifestations of neonatal thromboembolic events. The etiology of renal vein thrombosis remains unidentified in the majority of cases. We report a case of renal vein thrombosis in a neonate associated with elevated maternal lipoprotein (a). CASE PRESENTATION: A full-term female infant, appropriate for gestational age, was born via spontaneous vaginal delivery to an 18-year-old primigravida. The infant's birth weight was 3680 g and the Apgar scores were eight and nine at 1 and 5 minutes respectively. Evaluation of the infant in the newborn nursery revealed a palpable mass in the right lumbar area. Tests revealed hematuria and a high serum creatinine level of 1.5 mg/dl. An abdominal ultrasound Doppler flow study demonstrated an enlarged right kidney, right renal vein thrombosis, and progression of the thrombosis to the inferior vena cava. There was no evidence of saggital sinus thrombosis. An extensive work up of parents for hypercoagulable conditions was remarkable for a higher plasma lipoprotein (a) level of 73 mg/dl and an elevated fibrinogen level of 512 mg/dl in the mother. All paternal levels were normal. The plasma lipoprotein (a) level in the neonate was also normal. The neonate was treated with low molecular weight heparin (enoxaparin) at 1.5 mg/kg/day every 12 hours for 2 months, at which time a follow-up ultrasound Doppler flow study showed resolution of the thrombosis in both the renal vein and the inferior vena cava. CONCLUSION: There have been no studies to date that have explored the effect of abnormal maternal risk factors on fetal hemostasis. A case-control study is required to investigate whether elevated levels of maternal lipoprotein (a) may be a risk factor for neonatal thrombotic processes. Although infants with this presentation are typically treated with anticoagulation, there is a lack of evidence-based guidelines. Treatment modalities vary between study and treatment centers which warrants the establishment of a national registry. PMID- 18400109 TI - Microarray analysis of E9.5 reduced folate carrier (RFC1; Slc19a1) knockout embryos reveals altered expression of genes in the cubilin-megalin multiligand endocytic receptor complex. AB - BACKGROUND: The reduced folate carrier (RFC1) is an integral membrane protein and facilitative anion exchanger that mediates delivery of 5-methyltetrahydrofolate into mammalian cells. Adequate maternal-fetal transport of folate is necessary for normal embryogenesis. Targeted inactivation of the murine RFC1 gene results in post-implantation embryolethality, but daily folic acid supplementation of pregnant dams prolongs survival of homozygous embryos until mid-gestation. At E10.5 RFC1-/- embryos are developmentally delayed relative to wildtype littermates, have multiple malformations, including neural tube defects, and die due to failure of chorioallantoic fusion. The mesoderm is sparse and disorganized, and there is a marked absence of erythrocytes in yolk sac blood islands. The identification of alterations in gene expression and signaling pathways involved in the observed dysmorphology following inactivation of RFC1 mediated folate transport are the focus of this investigation. RESULTS: Affymetrix microarray analysis of the relative gene expression profiles in whole E9.5 RFC1-/- vs. RFC1+/+ embryos identified 200 known genes that were differentially expressed. Major ontology groups included transcription factors (13.04%), and genes involved in transport functions (ion, lipid, carbohydrate) (11.37%). Genes that code for receptors, ligands and interacting proteins in the cubilin-megalin multiligand endocytic receptor complex accounted for 9.36% of the total, followed closely by several genes involved in hematopoiesis (8.03%). The most highly significant gene network identified by Ingenuitytrade mark Pathway analysis included 12 genes in the cubilin-megalin multiligand endocytic receptor complex. Altered expression of these genes was validated by quantitative RT-PCR, and immunohistochemical analysis demonstrated that megalin protein expression disappeared from the visceral yolk sac of RFC1-/- embryos, while cubilin protein was widely misexpressed. CONCLUSION: Inactivation of RFC1 impacts the expression of several ligands and interacting proteins in the cubilin-amnionless-megalin complex that are involved in the maternal-fetal transport of folate and other nutrients, lipids and morphogens such as sonic hedgehog (Shh) and retinoids that play critical roles in normal embryogenesis. PMID- 18400111 TI - Ambulance clinical placements--a pilot study of students' experience. AB - BACKGROUND: Undergraduate paramedic students undertake clinical placements in a variety of locations. These placements are considered an essential element for paramedic pre-employment education. However, anecdotal evidence suggests some students have not had positive experiences on their emergency ambulance placements. The objective of this study was to identify the type of experiences had by students during ambulance clinical placements and to provide feedback to the ambulance services. METHODS: In this pilot study we employed a cross sectional study methodology, using a convenience sample of undergraduate paramedic students available in semester one of 2007 to ascertain the students' views on their reception by on-road paramedics and their overall experience on emergency ambulance clinical placements. Ethics approval was granted. RESULTS: There were 77 students who participated in the survey, 64% were females, with 92% of students < 25 years of age and 55% < 65 Kg in weight. There was a statistically significant difference in average height between the genders (Male 179 cm vs Female 168 cm, p < 0.001). Clinical instructors were available to 44% of students with 30% of students excluded from patient management. Thirty percent of students felt there was a lot of unproductive down time during the placement. Paramedics remarked to 40% of students that they doubted their ability to perform the physical role of a paramedic, of this group 36% were advised this more than once. CONCLUSION: This study demonstrates that for a small group of students, emergency ambulance clinical placements were not a positive experience clinically or educationally. Some qualified paramedics doubt if a number of female students can perform the physical role of a paramedic. PMID- 18400112 TI - Polymorphisms of Glutathione S-transferases Omega-1 among ethnic populations in China. AB - BACKGROUND: Glutathione S-transferases (GSTs) is a genetic factor for many diseases and exhibits great diversities among various populations. We assessed association of the genotypes of Glutathione S-transferases Omega-1 (GSTO1) A140D with ethnicity in China. RESULTS: Peripheral blood samples were obtained from 1314 individuals from 14 ethnic groups. Polymorphisms of GSTO1 A140D were measured using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Logistic regression was employed to adjustment for regional factor. The frequency of GSTO1 140A allele was 15.49% in the total 14 ethnic populations. Compared to Han ethnic group, two ethnic populations were more likely to have AA or CA genotype [odds ratio (OR): 1.77, 95% confidence interval (95% CI): 1.05 2.98 for Uygur and OR: 1.78, 95% CI: 1.18-2.69 for Hui]. However, there were no statistically significant differences across 14 ethnic groups when region factor was adjusted. In Han ethnicity, region was significantly associated with AA or CA genotype. Han individuals who resided in North-west of China were more likely to have these genotypes than those in South of China (OR: 1.63, 95% CI: 1.21-2.20). CONCLUSION: The prevalence of the GSTO1 140A varied significantly among different regional populations in China, which showed that geography played a more important role in the population differentiation for this allele than the ethnicity/race. PMID- 18400114 TI - Rapid reviews versus full systematic reviews: an inventory of current methods and practice in health technology assessment. AB - OBJECTIVES: This review assessed current practice in the preparation of rapid reviews by health technology assessment (HTA) organizations, both internationally and in the Australian context, and evaluated the available peer-reviewed literature pertaining to the methodology used in the preparation of these reviews. METHODS: A survey tool was developed and distributed to a total of fifty International Network of Agencies for Health Technology Assessment (INAHTA) members and other selected HTA organizations. Data on a broad range of themes related to the conduct of rapid reviews were collated, discussed narratively, and subjected to simple statistical analysis where appropriate. Systematic searches of the Cochrane Library, EMBASE, MEDLINE, and the Australian Medical Index were undertaken in March 2007 to identify literature pertaining to rapid review methodology. Comparative studies, guidelines, program evaluations, methods studies, commentaries, and surveys were considered for inclusion. RESULTS: Twenty three surveys were returned (46 percent), with eighteen agencies reporting on thirty-six rapid review products. Axiomatic trends were identified, but there was little cohesion between organizations regarding the contents, methods, and definition of a rapid review. The twelve studies identified by the systematic literature search did not specifically address the methodology underpinning rapid review; rather, many highlighted the complexity of the area. Authors suggested restricted research questions and truncated search strategies as methods to limit the time taken to complete a review. CONCLUSIONS: Rather than developing a formalized methodology by which to conduct rapid reviews, agencies should work toward increasing the transparency of the methods used for each review. It is perhaps the appropriate use, not the appropriate methodology, of a rapid review that requires future consideration. PMID- 18400113 TI - miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample. AB - BACKGROUND: MicroRNAs (miRNAs) are small endogenous non-coding interfering RNA molecules regarded as major regulators in eukaryotic gene expression. Different methods are employed for miRNA expression profiling. For a better understanding of their role in essential biological processes, convenient methods for differential miRNA expression analysis are required. RESULTS: Here, we present the miR-Q assay as a highly sensitive quantitative reverse transcription PCR (qRT PCR) for expression analysis of small RNAs such as miRNA molecules. It shows a high dynamic range of 6 to 8 orders of magnitude comprising a sensitivity of up to 0.2 fM miRNA, which corresponds to single copies per cell. There is nearly no cross reaction among closely-related miRNA family members, which points to the high specificity of the assays. Using this approach, we quantified the expression of let-7b in different human cell lines as well as miR-145 and miR-21 expression in porcine intestinal samples. CONCLUSION: miR-Q is a cost-effective and highly specific approach, which neither requires the use of fluorochromic probes, nor Locked Nucleic Acid (LNA)-modified oligonucleotides. Moreover, it provides a remarkable increase in specificity and simplified detection of small RNAs. PMID- 18400115 TI - Why licensing authorities need to consider the net value of new drugs in assigning review priorities: addressing the tension between licensing and reimbursement. AB - Pharmaceutical regulators and healthcare reimbursement authorities operate in different intellectual paradigms and adopt very different decision rules. As a result, drugs that have been licensed are often not available to all patients who could benefit because reimbursement authorities judge that the cost of therapies is greater than the health produced. This finding creates uncertainty for pharmaceutical companies planning their research and development investment, as licensing is no longer a guarantee of market access. In this study, we propose that it would be consistent with the objectives of pharmaceutical regulators to use the Net Benefit Framework of reimbursement authorities to identify those therapies that should be subject to priority review, that it is feasible to do so and that this would have several positive effects for patients, industry, and healthcare systems. PMID- 18400116 TI - Systematic reviews and economic evaluations conducted for the National Institute for Health and Clinical Excellence in the United Kingdom: a game of two halves? AB - BACKGROUND: Decision analytic models, as used in economic evaluations, require data on several clinical parameters. The gold standard approach is to conduct a systematic review of the relevant clinical literature, although reviews of economic evaluations indicate that this is rarely done. Technology appraisals for the National Institute for Health and Clinical Excellence (NICE), which are fully funded, represent the best case scenario for the close integration of economic evaluations and systematic reviews. The objective of this study was to assess the extent to which the systematic review of the clinical literature informs the economic evaluation in NICE technology appraisals. METHODS: All NICE technology assessment reports (TARs) published between January 2003 and July 2006 were considered. Data were abstracted on the TAR topics, the primary measure of clinical effectiveness, the approach to pooling in the clinical review, the measure of economic benefit and the use, or non-use, of the systematic review in the economic evaluation. RESULTS: Forty-one TARs were published in the period studied, all of which contained a systematic review. Most of the economic evaluations (85 percent) were cost-utility analyses, reflecting NICE's guidelines for economic evaluation. In seventeen cases, the clinical data were not pooled in the review, owing to heterogeneity in the clinical data or the limited number of studies. In these cases, the economists used alternative approaches for estimating the key effectiveness parameter in the model. The results of the review (when pooled) were always used when the primary clinical effectiveness measure corresponded with the measure of economic benefit (e.g., survival). However, because preference-based quality of life measures are rarely included in clinical trials, the results of the systematic review were never directly used in the cost-utility analyses. Nevertheless, the outputs of the systematic review were used when the data were useful in estimating components of the quality adjusted life-year (QALY) (e.g., the life-years gained, or the frequencies of health states to which QALYs could be assigned). Problems occurred mainly when the clinical data were not pooled, or when the measure of clinical benefit could not be converted into health states to which QALYs could be assigned. CONCLUSIONS: Economic evaluations can benefit from systematic reviews of the clinical literature. However, such reviews are not a panacea for conducting a good economic evaluation. Much of the relevant data for estimating QALYs are not contained in such reviews and the chosen method for summarizing the clinical data may inhibit the assessment of economic benefit. Problems would be reduced if those undertaking the technology assessments discussed the data requirements for the economic model at an early stage. PMID- 18400117 TI - Increasing decision-makers' access to economic evaluations: alternative methods of communicating the information. AB - OBJECTIVES: Although the importance of economic evaluations is recognized, research suggests the ways in which studies are summarized may not be optimal for a busy decision maker with little training in economics methodology. Therefore, the objective of this study was to seek decision makers' views on different summary formats, including a score, short summary, and structured abstracts of different degrees of detail. METHODS: We contacted 2,400 people, of which 84 decision makers volunteered and were presented, cumulatively, with different formats and asked whether these provided sufficient detail on the methodology and results of an economic study. RESULTS: From the fifty decision makers who responded to the questionnaire, it was found that the preferred combination was a very short summary, plus a more detailed structured abstract. It was also found that decision makers with economics training preferred the most detailed format, partly reflecting their reasons for consulting economic evaluations. CONCLUSIONS: Decision makers require both an initial screen of study content, plus more detail should they find the study relevant or interesting. PMID- 18400118 TI - Physician awareness of diagnostic and nondrug therapeutic costs: a systematic review. AB - OBJECTIVES: The aim of this study was to investigate doctors' knowledge of the relative and absolute costs of diagnostic tests, medical consumables (e.g., syringes or intravenous tubing), and healthcare visits as well as to determine factors influencing awareness. METHODS: For this systematic review, we searched the Cochrane Library, EconoLit, EMBASE, and MEDLINE; reviewed reference lists; and had contact with authors. Studies were included if either doctors or trainees were surveyed, there were >10 survey respondents, costs of diagnostic or therapeutic items were estimated, results were expressed quantitatively, and a clear description was provided of how authors defined Accurate Estimates and determined True Cost. Two authors reviewed each article for eligibility and extracted data independently. Cost accuracy outcomes were summarized, but data were not combined due to extensive heterogeneity. RESULTS: Fourteen articles were included in the final analysis. Cost accuracy was low; 33 percent of estimates were within 20 percent or 25 percent of true cost and 50 percent were within 50 percent or in the 50-200 percent range of the true cost. Country, year of study, level of training, and specialty did not impact accuracy. The cost of items appears to have no impact on the accuracy (Fisher's exact test, p = .41) or pattern of estimation (binomial test, p = .92). CONCLUSIONS: Doctors have a limited understanding of diagnostic and nondrug therapeutic costs, and we could not identify anything that impacts understanding of these costs. More focus is required in the education of physicians about costs and the access to cost information. PMID- 18400119 TI - Influence of expert clinical adviser characteristics on opinions about interventional procedures. AB - OBJECTIVES: The aim of this study was to examine potential associations between the professional background and experience of expert clinicians and their opinions about the clinical utility of interventional procedures. METHODS: A retrospective survey of expert clinician characteristics and their opinions was conducted. Information was collected on expert clinical adviser self-declared "operator," "researcher," and conflict of interest status. Associations were sought between expert clinical adviser characteristics and their opinions on whether procedures were "established," "efficacious," and "safe." The setting was the Interventional Procedures Programme of the UK's National Institute for Health and Clinical Excellence (NICE). A total of 598 expert clinician questionnaires relating to 182 different interventional procedures were analyzed. RESULTS: Expert clinical advisers with operative experience of procedures were significantly more likely to consider them as established (odds ratio [OR] 3.93; 95 percent confidence interval [CI], 2.43 to 6.36; p < .001), efficacious (OR 1.76; 95 percent CI, 1.00 to 3.08; p = .049), and safe (OR 2.28; 95 percent CI, 1.43 to 3.65; p = .001). Once adjusted for other characteristics, there was no association between either researcher or conflict of interest status and opinions about the clinical utility of procedures. CONCLUSIONS: Expert clinical advisers are an important source of information for decision makers producing guidance about the use of procedures, especially when published evidence is sparse or of poor quality. This study suggests that those who are operators, but not those who are researchers or declare a conflict of interest, are more likely to have a favorable opinion of a procedure's clinical utility. Use of expert clinical advisers with a variety of experience and backgrounds seems a reasonable approach to obtaining authoritative opinions about interventional procedures, to supplement and help interpret evidence from published data. PMID- 18400120 TI - Direct versus indirect comparisons: a summary of the evidence. AB - OBJECTIVES: The aim of this study was to provide a comprehensive summary and interpretation of the current evidence on the use and validity of statistical methods to conduct indirect comparisons of treatment effects. METHODS: A narrative review was conducted. RESULTS: Well-conducted methodological studies provide good evidence that adjusted indirect comparisons can lead to results similar to those from direct comparisons. The internal validity of several statistical methods to conduct indirect comparisons, therefore, has been established. Meta-regression, logistic regression, or adjusted indirect comparisons should be the methods of first choice. Unadjusted indirect comparisons are always unacceptable. Deciding whether to combine direct and indirect evidence will be a matter of informed judgment based on the similarities and dissimilarities of populations and interventions. Unverifiable assumptions with respect to the similarity of compared studies and low power are serious limitations of indirect comparisons. CONCLUSIONS: In the absence of sufficient head-to-head evidence, adjusted indirect comparisons can be viewed as additional analytical tools to determine the comparative efficacy and effectiveness of competing interventions. Researchers who use indirect comparisons need to keep the limitations in mind. PMID- 18400121 TI - Cost-effectiveness and budget impact of adjunctive hyperbaric oxygen therapy for diabetic foot ulcers. AB - BACKGROUND: Hyperbaric oxygen therapy (HBOT) has been proposed as an adjunct to standard methods of care for diabetic foot ulcers (DFU). Its use may decrease the risk of infection and lower extremity amputations (LEAs). As part of a Canadian assessment, we estimated the cost-effectiveness and budget impact of HBOT in this application. METHODS: We developed a decision model comparing adjunctive HBOT with standard care alone. The population was a 65-year-old cohort with DFU. The time horizon was 12 years taken from a Ministry of Health perspective. The health states were a healed wound with or without a minor LEA, an unhealed wound with no related surgery, and a major LEA. Efficacy data were based on outcomes reported in studies included in a literature review. Cost and capacity needs for treating DFU patients in Canada were estimated using prevalence data from the literature, and cost and utilization data from government records. RESULTS: The 12-year cost for patients receiving HBOT was CND$40,695 compared with CND$49,786 for standard care alone. Outcomes were 3.64 quality-adjusted life-years (QALYs) for those receiving HBOT and 3.01 QALYs for controls. Estimated cost to treat all prevalent DFU cases in Canada was CND$14.4-19.7 million/year over 4 years. If seven-person HBOT chambers were used, a further nineteen to thirty-five machines would be required nationally. CONCLUSIONS: Adjunctive HBOT for DFU is cost-effective compared with standard care. Additional HBOT capacity would be needed if it were to be adopted as the standard of care throughout Canada. PMID- 18400122 TI - Cost-effectiveness of primarily human papillomavirus-based cervical cancer screening in settings with currently established Pap screening: a systematic review commissioned by the German Federal Ministry of Health. AB - OBJECTIVES: The aim of this study was to summarize the current evidence for the cost-effectiveness of primarily human papillomavirus (HPV) -based cervical cancer screening in settings with already established Papanicolaou test (Pap) programs. Emphasis was placed on the German situation with annual Pap screening. METHODS: Medical, economic, and health technology assessment (HTA) databases were systematically searched for cost-effectiveness studies comparing HPV to Pap screening. Study data were extracted, standardized, and summarized in cost effectiveness plots contrasting HPV strategies to Pap screening with 1-, 2-, 3-, and 5-years interval. For each Pap setting, the likelihood of cost-effective HPV screening was assessed depending on willingness-to-pay. RESULTS: We reviewed twelve decision-analytic cost-effectiveness models. Study results showed wide variation due to methodical heterogeneity. Data synthesis revealed that the cost effectiveness of HPV screening depends on the interval of the established Pap screening strategy. In comparison with Pap screening every 2 years, only 25 percent of the HPV-based screening strategies were cost-effective. However, in comparison with Pap screening every 1, 3, or 5 years, 83 percent, 55 percent, and 92 percent of HPV screening strategies were cost-effective, respectively. Results for settings with annual Pap screening are based on models assuming 100 percent screening coverage. CONCLUSIONS: The introduction of HPV-based screening programs is cost-effective if the screening interval of the established Pap program exceeds 2 years. In settings with biennial Pap screening, introduction of HPV based screening is unlikely to be cost-effective. Results also suggest cost effectiveness of HPV-based screening in settings with annual Pap screening; however, this finding should be confirmed under realistic screening adherence assumptions. PMID- 18400123 TI - Cost-effectiveness of a multidisciplinary fall prevention program in community dwelling elderly people: a randomized controlled trial (ISRCTN 64716113). AB - OBJECTIVES: Multidisciplinary and multifactorial interventions seem to be effective in preventing falls. We aimed to assess the cost-effectiveness of a multidisciplinary fall prevention program compared with usual Dutch healthcare in community-dwelling people 65 years of age or older who experienced a fall. METHODS: Cost-effectiveness and cost-utility analysis were performed from a societal perspective. Falls and healthcare utilization were continuously measured for 12 months. Daily functioning and quality of life were measured at baseline, after 4 and 12 months. Bootstrap analyses were performed to estimate uncertainty of the findings and sensitivity analysis to assess the generalizability of assumptions made. RESULTS: One hundred sixty-six participants were randomly allocated to the experimental group and 167 to the control group. The overall response rate was 74 percent. Healthcare and patient and family costs of both groups were comparable. Our analyses showed no effect of the intervention program on falls, daily functioning, or quality of life measures. CONCLUSIONS: The multidisciplinary intervention program to prevent falls was not cost-effective compared with usual care in the Netherlands. Notwithstanding our findings, however, falls still have an important impact on society and individuals in terms of costs and effects. Economic evaluations studying promising interventions to prevent falls, therefore, remain necessary. PMID- 18400124 TI - Economic evaluation of screening for open-angle glaucoma. AB - OBJECTIVES: The aim of this study was to assess the cost-effectiveness of screening for open-angle glaucoma (OAG) in the United Kingdom, given that OAG is an important cause of blindness worldwide. METHODS: A Markov model was developed to estimate lifetime costs and benefits of a cohort of patients facing, alternatively, screening or current opportunistic case finding strategies. Strategies, varying in how screening would be organized (e.g., invitation for assessment by a glaucoma-trained optometrist [GO] or for simple test assessment by a technician) were developed, and allowed for the progression of OAG and treatment effects. Data inputs were obtained from systematic reviews. Deterministic and probabilistic sensitivity analyses were performed. RESULTS: Screening was more likely to be cost-effective as prevalence increased, for 40 year olds compared with 60 or 75 year olds, when the re-screening interval was greater (10 years), and for the technician strategy compared with the GO strategy. For each age cohort and at prevalence levels of < or =1 percent, the likelihood that either screening strategy would be more cost-effective than current practice was small. For those 40 years of age, "technician screening" compared with current practice has an incremental cost-effectiveness ratio (ICER) that society might be willing to pay when prevalence is 6 percent to 10 percent and at over 10 percent for 60 year olds. In the United Kingdom, the age specific prevalence of OAG is much lower. Screening by GO, at any age or prevalence level, was not associated with an ICER < pound 30,000. CONCLUSIONS: Population screening for OAG is unlikely to be cost-effective but could be for specific subgroups at higher risk. PMID- 18400125 TI - Assessing short-term effects and costs at an early stage of innovation: the use of positron emission tomography on radiotherapy treatment decision making. AB - OBJECTIVES: Positron emission tomography (PET) is an innovative imaging tool. Associated with computed tomography (CT), it allows a better definition for the tumor volume for radiotherapy, compared with CT only. The aim of this study was to assess the effects of PET on resource allocation (costs and savings) and on the choice of the following treatment in radiotherapy. METHODS: In 2004 and 2005, 209 patients were enrolled (97 patients with Hodgkin's disease and 112 with non small cell lung cancer) in a national study conducted in eight hospitals. Two treatment decisions made on the basis of CT only or CT associated with PET, were compared in a prospective study where each subject was his/her own control. The direct medical cost of using PET was assessed by microcosting, using data collected from specific questionnaires. The costs of new tests and the costs and savings associated with changes in the chosen treatment were calculated on the basis of reimbursement rates. RESULTS: The mean cost of using PET was approximately euro 800 per patient (50 percent for the radionuclide 18F-FDG [2 [18F]fluoro-2-deoxy-D-glucose]). Radiotherapy treatments were modified for 10 percent of patients with Hodgkin's disease versus 40 percent of patients with lung cancer. Overall, the use of PET induced both increases and decreases in the mean cost per patient: the net effect was a euro 425 and euro 931 cost increase in lung cancer and Hodgkin's disease, respectively. CONCLUSIONS: The use of PET for radiotherapy decision making seems more valuable for lung cancer than for Hodgkin's disease, both in terms of costs and changes in radiotherapy treatment. This result might help policy makers for prioritization. PMID- 18400126 TI - Extracapsular hip fracture management: cost-consequences analysis of two alternative operative methods. AB - OBJECTIVES: The aim of this prospective study was to perform a cost and outcome comparison between two alternative operative techniques (osteosynthesis and hemiarthroplasty) used in the treatment of elderly patients with unstable trochanteric hip fracture. MATERIALS AND METHODS: One hundred seventy-three trochanteric hip fracture patients were followed-up for 1 year after surgery. For each operative technique, hospital treatment's cost per patient was computed. Mortality and complication rate in-hospital and at specific time points after surgery were used as outcome measures. Patients' functional level before and after hip fracture was estimated according to their mobility and ability to perform basic and instrumental activities of daily living. RESULTS: The cost for patients undergoing osteosynthesis reached euro 1,931 per case, whereas for those treated with hemiarthroplasty reached euro 3,719 per case (2001 rates). There was no statistically significant difference regarding in-hospital mortality and complication rate, as well as mortality and complication rate 1 year after surgery, between the two patient groups. CONCLUSIONS: The quite similar performance of the two operative techniques suggests that cost could be the key factor for choosing between them. However, it is critical that many more randomized studies, with larger sample sizes and wider follow-up time periods should be conducted. PMID- 18400127 TI - Health-related quality of life in elderly patients with familial hypercholesterolemia. AB - OBJECTIVES: There are very few descriptions on how familial hypercholesterolemia (FH), a genetic disorder with an increased risk of coronary heart disease (CHD), affects older patients. The aim of this study was to analyze the health-related quality of life (HRQoL) of elderly patients with FH. METHODS: All FH patients with the same FH North Karelia-mutation aged 65 and over living in North Karelia province in Eastern Finland were asked to participate in the study in 2003. They were compared with age-standardized controls from the Finnish general population. A total of forty-three elderly FH patients were identified and thirty-seven of them (aged 65 to 84 years, M/F 10/27) agreed to participate. Most of them (27/37) had been diagnosed with CHD. All but one of these FH patients had been using statins for approximately 15 years. HRQoL was assessed with the RAND-36 (SF-36) and 15D instruments. RESULTS: Despite the clear cardiovascular morbidity and low socioeconomic status, these FH patients enjoyed a similar HRQoL as the age standardized controls in the general population. CONCLUSIONS: Lifelong hypercholesterolemia was not associated with poorer HRQoL in elderly survivors of FH. These elderly FH individuals were characterized by their healthy lifestyle and long-term statin treatment. PMID- 18400128 TI - Analysis of strengths, weaknesses, opportunities, and threats in the development of a health technology assessment program in Turkey. AB - OBJECTIVES: The Turkish healthcare system is currently undergoing reform, and efficient use of resources has become a key factor in determining the allocation of resources. The objective of this study was to analyze strengths, weaknesses, opportunities, and threats (SWOT) in the development of a health technology assessment (HTA) program in Turkey. METHODS: A SWOT analysis was performed using a literature review and interviews with key people in the Turkish Ministry of Health and Ministry of Labor and Social Security. RESULTS: Regarding recent reforms in health care, investments for information network and databank are the strengths, but the traditional "expert-based" decision making, poor availability of data, and poor quality of data could be seen as some of the weaknesses. Another major weakness is lack of general awareness of HTA. Increasing demand for transparency in decision making, demand for evidence, and demand for credibility by decision makers are some of the opportunities, and current healthcare reforms, i.e., restructuring of healthcare and general health insurance, could also be seen as major opportunities. These opportunities unfortunately could be threatened by lack of funding, and resources are challenged by large, recent national investments. CONCLUSIONS: There is a good opportunity for Turkey to use the skills in HTA currently being developed through activities in Europe and the Americas to assist in the development of a much more cost-effective and transparent healthcare system in Turkey. PMID- 18400129 TI - Comments to the manuscript by Martelli et al. in Volume 23:4. PMID- 18400131 TI - Metabotropic glutamate 5 receptors regulate sensitivity to ethanol in mice. AB - The metabotropic glutamate receptor 5 (mGlu5) has been implicated in ethanol- and drug-seeking behaviours in rodent studies. Here we examine a number of ethanol related behavioural assays in mice lacking mGlu5 and wild-type littermates. In a two-bottle free-choice paradigm, mGlu5-deficient mice consumed less ethanol with a reduced preference compared to wild-type mice. Indeed, mGlu5-deficienct mice were ethanol-avoiding at both concentrations of ethanol proffered (5% and 10% v/v). However, there was no difference in the rate of hepatic ethanol and acetaldehyde metabolism between genotypes and consumption of saccharin was similar. In a conditioned place preference study, mGlu5-deficient mice displayed a place preference for ethanol when conditioned with a low dose (1g/kg) of ethanol. Thus, while mGlu5-deficient mice consume less ethanol (with a reduced preference) than wild-type mice, this is not apparently related to impaired hepatic metabolism or a lack of reward from ethanol. Rather, we provide evidence that deletion of the mGlu5 receptor increases sensitivity to centrally mediated effects of ethanol. PMID- 18400133 TI - RNA interference: a new therapy for neuropathic pain? PMID- 18400134 TI - A rare cause of maternal death: liver and inferior vena cava rupture due to previously undiagnosed Ehlers-Danlos Syndrome type IV. PMID- 18400135 TI - Just a grommet: cardiac arrest in a healthy adolescent. PMID- 18400136 TI - Is time to peak effect of neuromuscular blocking agents dependent on dose? Testing the concept of buffered diffusion. AB - BACKGROUND AND OBJECTIVES: For neuromuscular blocking agents, an inverse relationship between potency and time to peak effect has been observed. To test the hypothesis that this relationship is due to buffered diffusion, we investigated the influence of dose on time to peak effect. Pharmacokinetic pharmacodynamic simulations were performed to support the expected relationships between potency, dose, peak effect and time to peak effect. METHODS: Pigs (20-28 kg body weight) were anaesthetized with ketamine and midazolam, followed by pentobarbital and fentanyl intravenously. Neuromuscular block was measured by stimulating the peroneal nerve supramaximally at 0.1 Hz and measuring the response of the tibialis anterior muscle mechanomyographically. After an initial dose to establish the individual ED90 of a neuromuscular blocking agent (rocuronium, vecuronium, pipecuronium or d-tubocurarine), five different doses of the same compound were administered to each animal, aiming at 20%, 40%, 60%, 75% or 90% block, in a random order. Doses were given 45 min after complete recovery of the twitch response. RESULTS: For rocuronium and pipecuronium, time to peak effect increased with dose, whereas dose did not affect time to peak effect of vecuronium and d-tubocurarine. Simulations predict that time to peak effect decreases with dose if buffered diffusion is taken into account. CONCLUSIONS: The results suggest that buffered diffusion does not play a dominant role in the time to peak effect of neuromuscular blocking agents. Therefore it is unlikely that the observed inverse relationship between potency and time to peak effect of neuromuscular blocking agents in the clinical range is due to buffered diffusion. PMID- 18400137 TI - Intraoperative epidural analgesia does not reduce time of postoperative analgesic treatment compared to intravenous analgesia. PMID- 18400138 TI - Subcutaneous hydration: a potentially hazardous route. PMID- 18400139 TI - Maternal factors implicated in fetal bradycardia after combined spinal epidural for labour pain. AB - BACKGROUND AND OBJECTIVE: Combined spinal epidural analgesia is effective for fast relief of severe labour pain but has been associated with worrisome decreases in fetal heart rate. Since the reasons for this phenomenon remain elusive, some anaesthesiologists may abstain from using this technique. We postulated that factors unrelated to the neuraxial technique could play a role in the decrease in fetal heart rate. To our knowledge, no prospective study has previously looked into this possibility. METHODS: We collected prospective data on 223 consecutive patients who received combined spinal epidural analgesia (123) or epidural analgesia (100). Maternal blood pressure, analogue pain scores, exogenous infusion of oxytocin, cervical dilatation, maternal age, parity and ethnicity were collected and correlated with the occurrence of decreases in fetal heart rate post combined spinal epidural. RESULTS: Univariate analysis showed a correlation between the incidence of fetal bradycardia and higher maternal pain scores, older maternal age, and combined spinal epidural analgesia. Multivariate analysis revealed that only pain scores and maternal age were independent predictors of fetal bradycardia post neuraxial blockade. CONCLUSIONS: Maternal pain scores and older maternal age are factors unrelated to the neuraxial technique that are independent predictors of fetal bradycardia after neuraxial analgesia for labour. PMID- 18400140 TI - Advantageous effects of dexmedetomidine on haemodynamic and recovery responses during extubation for intracranial surgery. AB - BACKGROUND AND OBJECTIVE: The anaesthetic method for intracranial neurosurgery must provide haemodynamic stability on emergence and allow early evaluation of the neurological status. In this study, we examined the effects of the alpha-2 agonist dexmedetomidine given at the end of the procedure to prevent hyperdynamic responses during extubation and to allow a comfortable and high-quality recovery. METHODS: Forty ASA I-III patients, aged between 18 and 75 yr, having elective intracranial surgery, were divided into two random groups. Standard procedures and drugs were used for monitoring, induction and maintenance. Isoflurane was reduced by 50% 5 min before the end of the surgery, and in Group I dexmedetomidine 0.5 microg kg(-1) and in Group II 20 mL of 0.9% NaCl were administrated intravenously over 60 s. Systolic, diastolic and mean arterial pressures, and heart rate were recorded before intravenous administration and also at 1, 3 and 5 min after administration, 1 min before extubation, during extubation, 1, 3, 5, 10, 15, 20 and 30 min after extubation. Duration of extubation and recovery were noted, and the quality of extubation was evaluated on a 5-point scale. RESULTS: Mean arterial pressure and heart rate were significantly higher in Group II than in Group I (P < 0.01). There were no statistically significant differences between groups regarding the duration of extubation and recovery (P > 0.05). Extubation quality score of all the patients were 1 in Group I and in Group II, the quality scores were 1 for 35%, 2 for 45% and 3 for 20% of the patients (P < 0.001). None of the patients in Group I and Group II showed respiratory depression, nausea or vomiting. CONCLUSION: Without interfering in recovery time, dexmedetomidine 0.5 microg kg(-1) administered 5 min before the end of surgery stabilizes haemodynamics, allows easy extubation, provides a more comfortable recovery and early neurological examination following intracranial operations. PMID- 18400141 TI - Comparison of dexmedetomidine-propofol vs. fentanyl-propofol for laryngeal mask insertion. AB - BACKGROUND AND OBJECTIVES: There have been many studies to find the optimum anaesthetics to provide excellent conditions for laryngeal mask insertion. We compared the effects of dexmedetomidine administered before propofol, on laryngeal mask insertion with fentanyl combined with propofol. METHODS: In all, 52 patients, ASA I-II, scheduled to have minor urological procedures were randomized into two groups. Group F received 1 microg kg(-1) fentanyl (in 10 mL normal saline) and Group D received 1 microg kg(-1) dexmedetomidine (in 10 mL normal saline). We used 1.5 mg kg(-1) propofol for induction and 50% N2O and 1.5% sevoflurane in oxygen for maintenance. We observed jaw mobility (1: fully relaxed; 2: mild resistance; 3: tight but opens; 4: closed), coughing or movement (1: none; 2: one or two coughs; 3: three or more coughs; 4: bucking/movement) and other events such as spontaneous ventilation, breath holding, expiratory stridor and lacrimation. In each category, scores <2 were acceptable for laryngeal mask insertion. RESULTS: More patients developed apnoea and their apnoea times were longer in Group F than Group D (P < 0.001). Respiratory rates increased in Group D (P < 0.001). Adverse events during laryngeal mask insertion were similar. The reductions in systolic and mean blood pressures were greater in Group F (systolic: P < 0.05, mean: P < 0.01). Emergence times were shorter in Group F than in Group D (P < 0.001). CONCLUSION: Dexmedetomidine, when used before propofol induction provides successful laryngeal mask insertion comparable to fentanyl, while preserving respiratory functions more than fentanyl. PMID- 18400142 TI - Remifentanil-TCI and propofol-TCI for conscious sedation during fibreoptic intubation in the acromegalic patient. AB - BACKGROUND AND OBJECTIVE: To evaluate the use of remifentanil-propofol administered as target-controlled infusion during awake fibreoptic intubation for anticipated difficult tracheal intubation in acromegalic patients. METHOD: In all, 20 consecutive acromegalic patients underwent elective endonasal endoscopic transsphenoidal pituitary surgery. After premedication with midazolam 0.03 mg kg( 1), initially a target-controlled infusion of remifentanil 1.0 ng mL(-1) and propofol 1.5 microg mL(-1) was started. The fibreoptic intubation was performed by the same physician experienced with the fibreoptic technique. During the fibreoptic procedure the target concentrations of remifentanil and propofol ranged between 1.0 and 5.0 ng mL(-1), and between 1.5 and 3.5 microg mL(-1), respectively. Changes in heart rate and mean arterial pressure were recorded during airway manipulation, during tracheal intubation, and at 1 and 3 min after. On the first postoperative day, patient recall and level of discomfort during fibreoptic intubation were evaluated. RESULTS: Endotracheal intubation was efficaciously and quickly secured in all patients. A significant increase in mean arterial pressure and heart rate was recorded only during tracheal intubation (P < 0.05). Oxygenation was sufficient and no bradypnea or apnoea was recorded. All patients later described their anaesthetic experience as satisfactory. During fibreoptic intubation, remifentanil (ng mL(-1)) and propofol (microg mL(-1)) mean effect-site concentrations were 3.2 +/- 0.3 and 2.0 +/- 1.0, respectively. CONCLUSION: Remifentanil and propofol target-controlled infusion provided satisfactory conscious sedation allowing for successful oral fibreoptic intubation in acromegalic patients with no recall. PMID- 18400144 TI - Pitfall in interpretation of TEG results. PMID- 18400143 TI - Evaluation of DNA damage in vivo induced by combined application of cisplatin and sevoflurane. AB - BACKGROUND AND OBJECTIVE: The influence of the combined application of cisplatin and sevoflurane on a variety of cell types of healthy mice or mice bearing Ehrlich ascites tumour has been investigated in an in vivo study. METHODS: The alkaline comet assay method was carried out on peripheral blood leucocytes, brain, liver, kidney and tumour cells of healthy mice or mice bearing Ehrlich ascites tumour. Groups of mice were treated intraperitoneally with cisplatin, exposed to sevoflurane or by combined treatment of sevoflurane after treatment with cisplatin for 3 consecutive days. RESULTS: The in vivo exposure to sevoflurane induced genotoxicity to all assayed cells. A strong synergistic genotoxic effect to peripheral blood leucocytes, liver and kidney cells was found in mice receiving both cisplatin and sevoflurane. In contrast, a decrease of the comet tail lengths of brain cells in the combined treatments was found as compared to cisplatin alone in both healthy (P < 0.001) and Ehrlich ascites tumour-bearing mice (P < 0.05), respectively. In addition, Ehrlich ascites tumour cells of mice treated with combined treatments showed a decrease in tail lengths (P < 0.001). These findings indicate an antagonistic effect of combined treatments. CONCLUSION: Treatment of mice with cisplatin and sevoflurane induced genotoxic effect in peripheral blood leucocytes, liver, kidney, brain and Ehrlich ascites tumour cells; synergistic effect of combined treatments was expressed in all cells but brain and Ehrlich ascites tumour cells. PMID- 18400145 TI - Cost-effectiveness of insulin aspart versus human soluble insulin in type 2 diabetes in four European countries: subgroup analyses from the PREDICTIVE study. AB - OBJECTIVES: To evaluate the long-term health economic outcomes associated with insulin aspart (IAsp) compared to human soluble insulin (HI) in type 2 diabetes patients on basal-bolus therapy in Sweden, Spain, Italy and Poland. METHODS: A published computer simulation model of diabetes was used to predict life expectancy, quality-adjusted life expectancy and incidence of diabetes-related complications. Baseline cohort characteristics (age 61.6 years, duration of diabetes 13.2 years, 45.1% male, HbA(1c) 8.2%, BMI 29.8 kg/m(2)) and treatment effects were derived from the PREDICTIVE observational study. Country-specific complication costs were derived from published sources. The analyses were run over 35-year time horizons from third-party payer perspectives in Spain, Italy and Poland and from a societal perspective in Sweden. Future costs and clinical benefits were discounted at country-specific discount rates. Sensitivity analyses were performed. RESULTS: IAsp was associated with improvements in discounted life expectancy and quality-adjusted life expectancy, and a reduced incidence of most diabetes-related complications versus HI in all four settings. IAsp was associated with societal cost-savings in Sweden (SEK 2470), direct medical cost savings in Sweden and Spain (SEK 8248 and euro 1382, respectively), but increased direct costs in Italy (euro 2235) and Poland (euro 743). IAsp was associated with improved quality-adjusted life expectancy in Sweden (0.077 QALYs), Spain (0.080 QALYs), Italy (0.120 QALYs) and Poland (0.003 QALYs). CONCLUSIONS: IAsp was dominant versus HI in both Sweden and Spain, would be considered cost-effective in Italy with an incremental cost-effectiveness ratio of euro 18,597 per QALY gained, but would not be considered cost-effective in Poland. PMID- 18400146 TI - Dosing accuracy of two insulin pre-filled pens. AB - OBJECTIVE: This study was designed to determine the dose accuracy of two commonly available insulin pre-filled pens for use in diabetes - FlexPen (Novo Nordisk A/S, Bagsvaerd, Denmark) (FP) and SoloStar (Sanofi-Aventis, Paris, France) (SS). RESEARCH DESIGN AND METHODS: Dosing accuracy was tested at 5 U, 10 U and 30 U doses - three previously unused pens of both pre-filled pens were used for each dose. At the 5 U dose each pen was tested 42 times, at the 10 U dose each pen was tested 25 times and at the 30 U dose each pen was tested 9 times. The pre-filled pens were used strictly according to manufacturers' instructions and measurements were made on a sensitive balance and corrected for the specific density of the insulin formulations used. Specified limits were based on ISO standards (+/-1 U for the 5 U and 10 U doses and +/-1.5 U for the 30 U dose). RESULTS: FP was more accurate for injecting 5 U, 10 U and 30 U doses (absolute mean 4.95 +/- 0.19 U, 9.61 +/- 0.27 U and 29.70 +/- 0.34 U, respectively) than SS (absolute mean 4.86 +/- 0.39 U, 9.27 +/- 0.52 U and 28.73 +/- 0.47 U, respectively). No doses were outside specified limits for 5 U and 30 U with FP and 1.3% of doses were outside these limits at the 10 U dose. For SS; 1.6%, 29.3% and 33.3% of doses were below the pre-specified threshold for the 5 U, 10 U and 30 U doses, respectively. CONCLUSIONS: This non-blinded comparison indicates that FP is a more accurate insulin pre-filled pen than SS at three different insulin doses. In this single user study, FP delivered consistent and accurate doses of insulin but SS had a high frequency of under dosing. No assessment was made of user variability in this study. PMID- 18400147 TI - Detection and prevalence patterns of group I coronaviruses in bats, northern Germany. AB - We tested 315 bats from 7 different bat species in northern Germany for coronaviruses by reverse transcription-PCR. The overall prevalence was 9.8%. There were 4 lineages of group I coronaviruses in association with 4 different species of verspertilionid bats (Myotis dasycneme, M. daubentonii, Pipistrellus nathusii, P. pygmaeus). The lineages formed a monophyletic clade of bat coronaviruses found in northern Germany. The clade of bat coronaviruses have a sister relationship with a clade of Chinese type I coronaviruses that were also associated with the Myotis genus (M. ricketti). Young age and ongoing lactation, but not sex or existing gravidity, correlated significantly with coronavirus detection. The virus is probably maintained on the population level by amplification and transmission in maternity colonies, rather than being maintained in individual bats. PMID- 18400148 TI - Exploring the assumptions underlying genetic variation in host nematode resistance (Open Access publication). AB - The wide range of genetic parameter estimates for production traits and nematode resistance in sheep obtained from field studies gives rise to much speculation. Using a mathematical model describing host - parasite interactions in a genetically heterogeneous lamb population, we investigated the consequence of: (i) genetic relationships between underlying growth and immunological traits on estimated genetic parameters for performance and nematode resistance, and (ii) alterations in resource allocation on these parameter estimates. Altering genetic correlations between underlying growth and immunological traits had large impacts on estimated genetic parameters for production and resistance traits. Extreme parameter values observed from field studies could only be reproduced by assuming genetic relationships between the underlying input traits. Altering preferences in the resource allocation had less pronounced effects on the genetic parameters for the same traits. Effects were stronger when allocation shifted towards growth, in which case worm burden and faecal egg counts increased and genetic correlations between these resistance traits and body weight became stronger. Our study has implications for the biological interpretation of field data, and for the prediction of selection response from breeding for nematode resistance. It demonstrates the profound impact that moderate levels of pleiotropy and linkage may have on observed genetic parameters, and hence on outcomes of selection for nematode resistance. PMID- 18400149 TI - Simultaneous fine mapping of closely linked epistatic quantitative trait loci using combined linkage disequilibrium and linkage with a general pedigree. AB - Causal mutations and their intra- and inter-locus interactions play a critical role in complex trait variation. It is often not easy to detect epistatic quantitative trait loci (QTL) due to complicated population structure requirements for detecting epistatic effects in linkage analysis studies and due to main effects often being hidden by interaction effects. Mapping their positions is even harder when they are closely linked. The data structure requirement may be overcome when information on linkage disequilibrium is used. We present an approach using a mixed linear model nested in an empirical Bayesian approach, which simultaneously takes into account additive, dominance and epistatic effects due to multiple QTL. The covariance structure used in the mixed linear model is based on combined linkage disequilibrium and linkage information. In a simulation study where there are complex epistatic interactions between QTL, it is possible to simultaneously map interacting QTL into a small region using the proposed approach. The estimated variance components are accurate and less biased with the proposed approach compared with traditional models. PMID- 18400150 TI - Genetic parameters related to environmental variability of weight traits in a selection experiment for weight gain in mice; signs of correlated canalised response. AB - Data from an experimental mice population selected from 18 generations to increase weight gain were used to estimate the genetic parameters associated with environmental variability. The analysis involved three traits: weight at 21 days, weight at 42 days and weight gain between 21 and 42 days. A dataset of 5273 records for males was studied. Data were analysed using Bayesian procedures by comparing the Deviance Information Criterion (DIC) value of two different models: one assuming homogeneous environmental variances and another assuming them as heterogeneous. The model assuming heterogeneity was better in all cases and also showed higher additive genetic variances and lower common environmental variances. The heterogeneity of residual variance was associated with systematic and additive genetic effects thus making reduction by selection possible. Genetic correlations between the additive genetic effects on mean and environmental variance of the traits analysed were always negative, ranging from -0.19 to 0.38. An increase in the heritability of the traits was found when considering the genetic determination of the environmental variability. A suggested correlated canalised response was found in terms of coefficient of variation but it could be insufficient to compensate for the scale effect associated with an increase of the mean. PMID- 18400151 TI - Data transformation for rank reduction in multi-trait MACE model for international bull comparison. AB - Since many countries use multiple lactation random regression test day models in national evaluations for milk production traits, a random regression multiple across-country evaluation (MACE) model permitting a variable number of correlated traits per country should be used in international dairy evaluations. In order to reduce the number of within country traits for international comparison, three different MACE models were implemented based on German daughter yield deviation data and compared to the random regression MACE. The multiple lactation MACE model analysed daughter yield deviations on a lactation basis reducing the rank from nine random regression coefficients to three lactations. The lactation breeding values were very accurate for old bulls, but not for the youngest bulls with daughters with short lactations. The other two models applied principal component analysis as the dimension reduction technique: one based on eigenvalues of a genetic correlation matrix and the other on eigenvalues of a combined lactation matrix. The first one showed that German data can be transformed from nine traits to five eigenfunctions without losing much accuracy in any of the estimated random regression coefficients. The second one allowed performing rank reductions to three eigenfunctions without having the problem of young bulls with daughters with short lactations. PMID- 18400152 TI - Exploration of cytoplasmic inheritance as a contributor to maternal effects in Welsh Mountain sheep. AB - Cytoplasmic effects were investigated using a dataset comprising three breeding groups of Welsh Mountain sheep. The influences of cytoplasmic effects were investigated by comparing animal models with and without a random term representing cytoplasmic effects. The models were applied to the eight-week weight, scan weight (mean 152 days) and ultrasonically scanned muscle and fat depth. The animal model included the random effects of animals and the maternal additive genetic, maternal permanent environmental and maternal common environmental effects. In total there were 24 569, 10 509, 8389, 8369 records for the eight-week weight, scan weight, muscle depth and fat depth respectively. Four subsets were further analysed containing maternal lines with at least five, ten, fifteen and twenty animals/line. There was no evidence of cytoplasmic effects on eight-week weight and muscle depth. Cytoplasmic effects contributed 1-2% of phenotypic variance for scan-weight and fat depth, but the effect was generally non-significant (P >0.05). As the number of animals per maternal line increased, the magnitude of cytoplasmic effects also increased for these traits. Direct heritability estimates for the eight-week weight, scan weight, muscle depth and fat depth using the full dataset were 0.18, 0.25, 0.24, and 0.21 respectively. PMID- 18400153 TI - Genetic differentiation between the Old and New types of Serbian Tsigai sheep. AB - Two Tsigai sheep populations exist in Serbia: the Old type, called Cokan, and the New type. It is assumed that the New type results from upgrading Tsigai sheep with exotic genetic material. We investigated genetic diversity and differentiation of these types by analysing 23 autosomal microsatellites. Tests for Hardy-Weinberg proportions, linkage equilibrium between genotypes across loci and the calculation of inbreeding coefficients were performed and the deficiency in the number of alleles within the Tsigai types was examined using a Wilcoxon sign-rank test. The New type displayed a higher level of genetic variability than the Cokan in terms of allele numbers, but the New Tsigai showed a pattern of heterozygosity deficiency. The positive f value for the Cokan suggests the occurrence of inbreeding in this type. The proportion of linkage disequilibrium was below that expected by chance. Exclusion of two loci in Hardy-Weinberg disequilibrium did not alter our conclusions based on the entire data set i.e. the two Tsigai types are clearly differentiated and the New Tsigai type has been influenced by crossbreeding. Therefore, the Cokan Tsigai should be considered as a distinct endangered breed in the FAO classification. PMID- 18400154 TI - Of cartridges and columns: new roles for cadherins in visual system development. AB - In this issue of Neuron, two papers provide new insights into roles for local cadherin-based interactions during axon targeting in the Drosophila visual system. Using high-resolution analyses, Chen and Clandinin identify nonautonomous roles for the atypical cadherin Flamingo during photoreceptor targeting, while Nern et al. demonstrate that local cell type-specific roles for N-cadherin control layer-specific targeting of lamina neurons. PMID- 18400155 TI - Neurogenesis or neuronal specification: phosphorylation strikes again! AB - How basic helix-loop-helix (bHLH) transcription factors control neurogenesis and neuronal subtype specification through transcriptional mechanisms mediated by cell signaling remains to be fully elucidated. In this issue of Neuron, Ma et al. discover that phosphorylation via GSK3 of the bHLH factor, Ngn2 (Neurog2), adds a neuronal subtype-specific program to its functional repertoire that is activated in the developing neural tube in vivo. PMID- 18400156 TI - Methamphetamine induces chronic corticostriatal depression: too much of a bad thing. AB - Leading theories of drug addiction propose that repeated drug exposure produces a long-lasting homeostatic dysregulation in brain reward processing that is normalized by drug readministration. In this issue of Neuron, Bamford and colleagues describe a novel neurobiological substrate that may contribute to this effect. PMID- 18400157 TI - A remarkable facilitating effect of parietal damage. AB - When distracters conflict with our instructions, our reactions normally get slower. However, Coulthard and colleagues in this issue of Neuron show that damage to the right parietal lobe reverses this effect, paradoxically facilitating responses. This surprising result may shed light on the functional role of parietal cortex within a larger cortical circuit for voluntary behavior. PMID- 18400158 TI - Paths of convergence: sirtuins in aging and neurodegeneration. AB - Members of the sirtuin family of protein deacetylases support and promote longevity in diverse organisms and can extend life span when upregulated. Sirtuin pathways also modulate fundamental mechanisms in aging-related neurodegenerative diseases, including protein aggregation, stress responses, mitochondrial homeostasis, and inflammatory processes. In this minireview, we will discuss how progress in understanding the neurobiology of sirtuins is shedding light on the pathogenesis of these devastating conditions. We will also examine the potential and challenges of targeting sirtuin pathways therapeutically. PMID- 18400159 TI - One-dimensional dynamics of attention and decision making in LIP. AB - Where we allocate our visual spatial attention depends upon a continual competition between internally generated goals and external distractions. Recently it was shown that single neurons in the macaque lateral intraparietal area (LIP) can predict the amount of time a distractor can shift the locus of spatial attention away from a goal. We propose that this remarkable dynamical correspondence between single neurons and attention can be explained by a network model in which generically high-dimensional firing-rate vectors rapidly decay to a single mode. We find direct experimental evidence for this model, not only in the original attentional task, but also in a very different task involving perceptual decision making. These results confirm a theoretical prediction that slowly varying activity patterns are proportional to spontaneous activity, pose constraints on models of persistent activity, and suggest a network mechanism for the emergence of robust behavioral timing from heterogeneous neuronal populations. PMID- 18400160 TI - The cadherin Flamingo mediates level-dependent interactions that guide photoreceptor target choice in Drosophila. AB - Quantitative differences in cadherin activity have been proposed to play important roles in patterning connections between pre- and postsynaptic neurons. However, no examples of such a function have yet been described, and the mechanisms that would allow such differences to direct growth cones to specific synaptic targets are unknown. In the Drosophila visual system, photoreceptors are genetically programmed to make a complex, stereotypic set of synaptic connections. Here we show that the atypical cadherin Flamingo functions as a short-range, homophilic signal, passing between specific R cell growth cones to influence their choice of postsynaptic partners. We find that individual growth cones are sensitive to differences in Flamingo activity through opposing interactions between neighboring cells and require these interactions to be balanced in order to extend along the appropriate trajectory. PMID- 18400161 TI - Local N-cadherin interactions mediate distinct steps in the targeting of lamina neurons. AB - The organization of neuronal processes into a series of layers is a hallmark of many brain regions. Homophilic cell adhesion molecules of the cadherin family have been implicated in layer choice. How they contribute to the targeting of neurons to distinct layers remains unclear. Here we systematically explore the role of a classical cadherin, Drosophila N-cadherin (CadN), in the targeting of five classes of related neurons to a series of consecutive layers in the fly visual system. We show that CadN is required in lamina neurons at discrete developmental steps but not used in a layer-specific fashion. Local CadN expression patterns correlate with specific growth cone movements, and CadN expression on one growth cone in a specific layer is essential for the targeting of processes of another neuron to this layer. We propose that dynamic regulation of CadN enables this widely expressed protein to mediate specific local interactions during neural circuit assembly. PMID- 18400162 TI - Endocytosis is required for synaptic activity-dependent release of amyloid-beta in vivo. AB - Aggregation of amyloid-beta (Abeta) peptide into soluble and insoluble forms within the brain extracellular space is central to the pathogenesis of Alzheimer's disease. Full-length amyloid precursor protein (APP) is endocytosed from the cell surface into endosomes where it is cleaved to produce Abeta. Abeta is subsequently released into the brain interstitial fluid (ISF). We hypothesized that synaptic transmission results in more APP endocytosis, thereby increasing Abeta generation and release into the ISF. We found that inhibition of clathrin mediated endocytosis immediately lowers ISF Abeta levels in vivo. Two distinct methods that increased synaptic transmission resulted in an elevation of ISF Abeta levels. Inhibition of endocytosis, however, prevented the activity dependent increase in Abeta. We estimate that approximately 70% of ISF Abeta arises from endocytosis-associated mechanisms, with the vast majority of this pool also dependent on synaptic activity. These findings have implications for AD pathogenesis and may provide insights into therapeutic intervention. PMID- 18400163 TI - Oscillations in notch signaling regulate maintenance of neural progenitors. AB - Expression of the Notch effector gene Hes1 is required for maintenance of neural progenitors in the embryonic brain, but persistent and high levels of Hes1 expression inhibit proliferation and differentiation of these cells. Here, by using a real-time imaging method, we found that Hes1 expression dynamically oscillates in neural progenitors. Furthermore, sustained overexpression of Hes1 downregulates expression of proneural genes, Notch ligands, and cell cycle regulators, suggesting that their proper expression depends on Hes1 oscillation. Surprisingly, the proneural gene Neurogenin2 (Ngn2) and the Notch ligand Delta like1 (Dll1) are also expressed in an oscillatory manner by neural progenitors, and inhibition of Notch signaling, a condition known to induce neuronal differentiation, leads to downregulation of Hes1 and sustained upregulation of Ngn2 and Dll1. These results suggest that Hes1 oscillation regulates Ngn2 and Dll1 oscillations, which in turn lead to maintenance of neural progenitors by mutual activation of Notch signaling. PMID- 18400164 TI - Regulation of motor neuron specification by phosphorylation of neurogenin 2. AB - The mechanisms by which proneural basic helix-loop-helix (bHLH) factors control neurogenesis have been characterized, but it is not known how they specify neuronal cell-type identity. Here, we provide evidence that two conserved serine residues on the bHLH factor neurogenin 2 (Ngn2), S231 and S234, are phosphorylated during motor neuron differentiation. In knockin mice in which S231 and S234 of Ngn2 were mutated to alanines, neurogenesis occurs normally, but motor neuron specification is impaired. The phosphorylation of Ngn2 at S231 and S234 facilitates the interaction of Ngn2 with LIM homeodomain transcription factors to specify motor neuron identity. The phosphorylation-dependent cooperativity between Ngn2 and homeodomain transcription factors may be a general mechanism by which the activities of bHLH and homeodomain proteins are temporally and spatially integrated to generate the wide diversity of cell types that are a hallmark of the nervous system. PMID- 18400165 TI - Setting clock speed in mammals: the CK1 epsilon tau mutation in mice accelerates circadian pacemakers by selectively destabilizing PERIOD proteins. AB - The intrinsic period of circadian clocks is their defining adaptive property. To identify the biochemical mechanisms whereby casein kinase1 (CK1) determines circadian period in mammals, we created mouse null and tau mutants of Ck1 epsilon. Circadian period lengthened in CK1epsilon-/-, whereas CK1epsilon(tau/tau) shortened circadian period of behavior in vivo and suprachiasmatic nucleus firing rates in vitro, by accelerating PERIOD-dependent molecular feedback loops. CK1epsilon(tau/tau) also accelerated molecular oscillations in peripheral tissues, revealing its global role in circadian pacemaking. CK1epsilon(tau) acted by promoting degradation of both nuclear and cytoplasmic PERIOD, but not CRYPTOCHROME, proteins. Together, these whole-animal and biochemical studies explain how tau, as a gain-of-function mutation, acts at a specific circadian phase to promote degradation of PERIOD proteins and thereby accelerate the mammalian clockwork in brain and periphery. PMID- 18400167 TI - Transgenic mice lacking NMDAR-dependent LTD exhibit deficits in behavioral flexibility. AB - While most studies have focused on the role of long-term potentiation in behavior, far less is known about the role of long-term depression (LTD). To examine the potential involvement of LTD in learning and memory, we generated transgenic mice that express a fragment of the SV40 small t antigen known to inhibit protein phosphatase 2A (PP2A). Small t antigen expression blocked both stimulus-induced and chemically induced NMDAR-dependent LTD at Schaffer collateral synapses but did not affect potentiation, depotentiation, or mGluR dependent LTD. This physiological phenotype was associated with deficits in behavioral flexibility in both the Morris water maze and a delayed nonmatch to place T-maze task, suggesting that NMDAR-dependent LTD is required for behavioral flexibility and may act by weakening previously encoded memory traces when new information is learned. PMID- 18400166 TI - Repeated exposure to methamphetamine causes long-lasting presynaptic corticostriatal depression that is renormalized with drug readministration. AB - Addiction-associated behaviors such as drug craving and relapse are hypothesized to result from synaptic changes that persist long after withdrawal and are renormalized by drug reinstatement, although such chronic synaptic effects have not been identified. We report that exposure to the dopamine releaser methamphetamine for 10 days elicits a long-lasting (>4 month) depression at corticostriatal terminals that is reversed by methamphetamine readministration. Both methamphetamine-induced chronic presynaptic depression and the drug's selective renormalization in drug-experienced animals are independent of corresponding long-term changes in synaptic dopamine release but are due to alterations in D1 dopamine and cholinergic receptor systems. These mechanisms might provide a synaptic basis that underlies addiction and habit learning and their long-term maintenance. PMID- 18400168 TI - Decoupling through synchrony in neuronal circuits with propagation delays. AB - The level of synchronization in distributed systems is often controlled by the strength of the interactions between individual elements. In brain circuits the connection strengths between neurons are modified under the influence of spike timing-dependent plasticity (STDP) rules. Here we show that when recurrent networks with conduction delays exhibit population bursts, STDP rules exert a strong decoupling force that desynchronizes activity. Conversely, when activity in the network is random, the same rules can have a coupling and synchronizing influence. The presence of these opposing forces promotes the self-organization of spontaneously active neuronal networks to a state at the border between randomness and synchrony. The decoupling force of STDP may be engaged by the synchronous bursts occurring in the hippocampus during slow-wave sleep, leading to the selective erasure of information from hippocampal circuits as memories are established in neocortical areas. PMID- 18400169 TI - Lateral sharpening of cortical frequency tuning by approximately balanced inhibition. AB - Cortical inhibition plays an important role in shaping neuronal processing. The underlying synaptic mechanisms remain controversial. Here, in vivo whole-cell recordings from neurons in the rat primary auditory cortex revealed that the frequency tuning curve of inhibitory input was broader than that of excitatory input. This results in relatively stronger inhibition in frequency domains flanking the preferred frequencies of the cell and a significant sharpening of the frequency tuning of membrane responses. The less selective inhibition can be attributed to a broader bandwidth and lower threshold of spike tonal receptive field of fast-spike inhibitory neurons than nearby excitatory neurons, although both types of neurons receive similar ranges of excitatory input and are organized into the same tonotopic map. Thus, the balance between excitation and inhibition is only approximate, and intracortical inhibition with high sensitivity and low selectivity can laterally sharpen the frequency tuning of neurons, ensuring their highly selective representation. PMID- 18400171 TI - NotI is not boring. PMID- 18400170 TI - Control over conflict during movement preparation: role of posterior parietal cortex. AB - Flexible behavior in humans often requires that rapid choices be made between conflicting action plans. Although much attention has focused on prefrontal regions, little is understood about the contribution of parietal cortex under situations of response conflict. Here we show that right parietal damage associated with spatial neglect leads to paradoxical facilitation (speeding) of rightward movements in the presence of conflicting leftward response plans. These findings indicate a critical role for parietal regions in action planning when there is response competition. In contrast, patients with prefrontal damage have an augmented cost of conflict for both leftward and rightward movements. The results suggest involvement of two independent systems in situations of response conflict, with right parietal cortex being a crucial site for automatic activation of competing motor plans and prefrontal regions acting independently to inhibit action plans irrelevant to current task goals. PMID- 18400172 TI - Exit biology: battle for the nascent chain. PMID- 18400173 TI - Characterization of a trifunctional mimivirus mRNA capping enzyme and crystal structure of the RNA triphosphatase domain. AB - The RNA triphosphatase (RTPase) components of the mRNA capping apparatus are a bellwether of eukaryal taxonomy. Fungal and protozoal RTPases belong to the triphosphate tunnel metalloenzyme (TTM) family, exemplified by yeast Cet1. Several large DNA viruses encode metal-dependent RTPases unrelated to the cysteinyl-phosphatase RTPases of their metazoan host organisms. The origins of DNA virus RTPases are unclear because they are structurally uncharacterized. Mimivirus, a giant virus of amoeba, resembles poxviruses in having a trifunctional capping enzyme composed of a metal-dependent RTPase module fused to guanylyltransferase (GTase) and guanine-N7 methyltransferase domains. The crystal structure of mimivirus RTPase reveals a minimized tunnel fold and an active site strikingly similar to that of Cet1. Unlike homodimeric fungal RTPases, mimivirus RTPase is a monomer. The mimivirus TTM-type RTPase-GTase fusion resembles the capping enzymes of amoebae, providing evidence that the ancestral large DNA virus acquired its capping enzyme from a unicellular host. PMID- 18400174 TI - Structure prediction of domain insertion proteins from structures of individual domains. AB - Multidomain proteins continue to be a major challenge in protein structure prediction. Here we present a Monte Carlo (MC) algorithm, implemented within Rosetta, to predict the structure of proteins in which one domain is inserted into another. Three MC moves combine rigid-body and loop movements to search the constrained conformation by structure disruption and subsequent repair of chain breaks. Local searches find that the algorithm samples and recovers near-native structures consistently. Further global searches produced top-ranked structures within 5 A in 31 of 50 cases in low-resolution mode, and refinement of top-ranked low-resolution structures produced models within 2 A in 21 of 50 cases. Rigid body orientations were often correctly recovered despite errors in linker conformation. The algorithm is broadly applicable to de novo structure prediction of both naturally occurring and engineered domain insertion proteins. PMID- 18400175 TI - Multiple states of a nucleotide-bound group 2 chaperonin. AB - Chaperonin action is controlled by cycles of nucleotide binding and hydrolysis. Here, we examine the effects of nucleotide binding on an archaeal group 2 chaperonin. In contrast to the ordered apo state of the group 1 chaperonin GroEL, the unliganded form of the homo-16-mer Methanococcus maripaludis group 2 chaperonin is very open and flexible, with intersubunit contacts only in the central double belt of equatorial domains. The intermediate and apical domains are free of contacts and deviate significantly from the overall 8-fold symmetry. Nucleotide binding results in three distinct, ordered 8-fold symmetric conformations--open, partially closed, and fully closed. The partially closed ring encloses a 40% larger volume than does the GroEL-GroES folding chamber, enabling it to encapsulate proteins up to 80 kDa, in contrast to the fully closed form, whose cavities are 20% smaller than those of the GroEL-GroES chamber. PMID- 18400176 TI - Structure of the mammalian 80S ribosome at 8.7 A resolution. AB - In this paper, we present a structure of the mammalian ribosome determined at approximately 8.7 A resolution by electron cryomicroscopy and single-particle methods. A model of the ribosome was created by docking homology models of subunit rRNAs and conserved proteins into the density map. We then modeled expansion segments in the subunit rRNAs and found unclaimed density for approximately 20 proteins. In general, many conserved proteins and novel proteins interact with expansion segments to form an integrated framework that may stabilize the mature ribosome. Our structure provides a snapshot of the mammalian ribosome at the beginning of translation and lends support to current models in which large movements of the small subunit and L1 stalk occur during tRNA translocation. Finally, details are presented for intersubunit bridges that are specific to the eukaryotic ribosome. We suggest that these bridges may help reset the conformation of the ribosome to prepare for the next cycle of chain elongation. PMID- 18400177 TI - Structures of the rare-cutting restriction endonuclease NotI reveal a unique metal binding fold involved in DNA binding. AB - The structure of the rare-cutting restriction endonuclease NotI, which recognizes the 8 bp target 5'-GCGGCCGC-3', has been solved with and without bound DNA. Because of its specificity (recognizing a site that occurs once per 65 kb), NotI is used to generate large genomic fragments and to map DNA methylation status. NotI contains a unique metal binding fold, found in a variety of putative endonucleases, occupied by an iron atom coordinated within a tetrahedral Cys4 motif. This domain positions nearby protein elements for DNA recognition, and serves a structural role. While recognition of the central six base pairs of the target is accomplished via a saturated hydrogen bond network typical of restriction enzymes, the most peripheral base pairs are engaged in a single direct contact in the major groove, reflecting reduced pressure to recognize those positions. NotI may represent an evolutionary intermediate between mobile endonucleases (which recognize longer target sites) and canonical restriction endonucleases. PMID- 18400178 TI - The evolutionarily conserved family of cyanovirin-N homologs: structures and carbohydrate specificity. AB - Solution structures for three members of the recently discovered cyanovirin-N (CV N) homolog family of lectins have been determined. Cyanovirin-N homologs (CVNHs) from Tuber borchii, Ceratopteris richardii, and Neurospora crassa, representing each of the three phylogenetic groups, were selected. All proteins exhibit the same fold, and the overall structures resemble that of the founding member of the family, CV-N, albeit with noteworthy differences in loop conformation and detailed local structure. Since no data are available regarding the proteins' function or their natural ligands, extensive carbohydrate-binding studies were conducted. We delineated ligand-binding sites on all three proteins by nuclear magnetic resonance and identified which sugars interact by array screening. The number and location of binding sites vary for the three proteins, and different ligand specificities exist. Potential physiological roles for two family members, TbCVNH and NcCVNH, were probed in nutrition deprivation experiments that suggest a possible involvement of these proteins in lifestyle-related responses. PMID- 18400179 TI - The GTPase cycle of the chloroplast import receptors Toc33/Toc34: implications from monomeric and dimeric structures. AB - Transport of precursor proteins across chloroplast membranes involves the GTPases Toc33/34 and Toc159 at the outer chloroplast envelope. The small GTPase Toc33/34 can homodimerize, but the regulation of this interaction has remained elusive. We show that dimerization is independent of nucleotide loading state, based on crystal structures of dimeric Pisum sativum Toc34 and monomeric Arabidopsis thaliana Toc33. An arginine residue is--in the dimer--positioned to resemble a GAP arginine finger. However, GTPase activation by dimerization is sparse and active site features do not explain catalysis, suggesting that the homodimer requires an additional factor as coGAP. Access to the catalytic center and an unusual switch I movement in the dimeric structure support this finding. Potential binding sites for interactions within the Toc translocon or with precursor proteins can be derived from the structures. PMID- 18400180 TI - Crystal structure of human factor VIII: implications for the formation of the factor IXa-factor VIIIa complex. AB - Factor VIII is a procofactor that plays a critical role in blood coagulation, and is missing or defective in hemophilia A. We determined the X-ray crystal structure of B domain-deleted human factor VIII. This protein is composed of five globular domains and contains one Ca(2+) and two Cu(2+) ions. The three homologous A domains form a triangular heterotrimer where the A1 and A3 domains serve as the base and interact with the C2 and C1 domains, respectively. The structurally homologous C1 and C2 domains reveal membrane binding features. Based on biochemical studies, a model of the factor IXa-factor VIIIa complex was constructed by in silico docking. Factor IXa wraps across the side of factor VIII, and an extended interface spans the factor VIII heavy and light chains. This model provides insight into the activation of factor VIII and the interaction of factor VIIIa with factor IXa on the membrane surface. PMID- 18400182 TI - Coarse-grained MD simulations of membrane protein-bilayer self-assembly. AB - Complete determination of a membrane protein structure requires knowledge of the protein position within the lipid bilayer. As the number of determined structures of membrane proteins increases so does the need for computational methods which predict their position in the lipid bilayer. Here we present a coarse-grained molecular dynamics approach to lipid bilayer self-assembly around membrane proteins. We demonstrate that this method can be used to predict accurately the protein position in the bilayer for membrane proteins with a range of different sizes and architectures. PMID- 18400181 TI - Crystal structure of the CaV2 IQ domain in complex with Ca2+/calmodulin: high resolution mechanistic implications for channel regulation by Ca2+. AB - Calmodulin (CaM) regulation of Ca(2+) channels is central to Ca(2+) signaling. Ca(V)1 versus Ca(V)2 classes of these channels exhibit divergent forms of regulation, potentially relating to customized CaM/IQ interactions among different channels. Here we report the crystal structures for the Ca(2+)/CaM IQ domains of both Ca(V)2.1 and Ca(V)2.3 channels. These highly similar structures emphasize that major CaM contacts with the IQ domain extend well upstream of traditional consensus residues. Surprisingly, upstream mutations strongly diminished Ca(V)2.1 regulation, whereas downstream perturbations had limited effects. Furthermore, our Ca(V)2 structures closely resemble published Ca(2+)/CaM Ca(V)1.2 IQ structures, arguing against Ca(V)1/2 regulatory differences based solely on contrasting CaM/IQ conformations. Instead, alanine scanning of the Ca(V)2.1 IQ domain, combined with structure-based molecular simulation of corresponding CaM/IQ binding energy perturbations, suggests that the C lobe of CaM partially dislodges from the IQ element during channel regulation, allowing exposed IQ residues to trigger regulation via isoform-specific interactions with alternative channel regions. PMID- 18400183 TI - Infinite kinetic stability against dissociation of supramolecular protein complexes through donor strand complementation. AB - Adhesive type 1 pili from uropathogenic Escherichia coli strains are heat and denaturant resistant, filamentous protein complexes. Individual pilus subunits associate through "donor strand complementation," whereby the incomplete immunoglobulin-like fold of each subunit is completed by the N-terminal extension of a neighboring subunit. We show that antiparallel donor strand insertion generally causes nonequilibrium behavior in protein folding and extreme activation energy barriers for dissociation of subunit-subunit complexes. We identify the most kinetically stable, noncovalent protein complex known to date. The complex between the pilus subunit FimG and the donor strand peptide of the subunit FimF shows an extrapolated dissociation half-life of 3 x 10(9) years. The 15 residue peptide forms ideal intermolecular beta sheet H-bonds with FimG over 10 residues, and its hydrophobic side chains strongly interact with the hydrophobic core of FimG. The results show that kinetic stability and nonequilibrium behavior in protein folding confers infinite stability against dissociation in extracellular protein complexes. PMID- 18400185 TI - The Roaring Twenties. AB - New cell types and cytokines have emerged as key participants in the elaboration of and recovery from inflammation. A collection of reviews covers recent advances in our understanding of this crucial component of host defense. PMID- 18400184 TI - Structural basis of site-specific histone recognition by the bromodomains of human coactivators PCAF and CBP/p300. AB - Histone lysine acetylation is central to epigenetic control of gene transcription. Bromodomains of chromosomal proteins function as acetyl-lysine (Kac) binding domains. However, how bromodomains recognize site-specific histones remains unanswered. Here, we report three three-dimensional solution structures of the bromodomains of the human transcriptional coactivators CREB-binding protein (CBP) and p300/CBP-associated factor (PCAF) bound to peptides derived from histone acetylation sites at lysines 36 and 9 in H3, and lysine 20 in H4. From structural and biochemical binding analyses, we determine consensus histone recognition by the bromodomains of PCAF and CBP, which represent two different subgroups of the bromodomain family. Through bromodomain residues in the ZA and BC loops, PCAF prefers acetylation sites with a hydrophobic residue at (Kac+2) position and a positively charged or aromatic residue at (Kac+3), whereas CBP favors bulky hydrophobic residues at (Kac+1) and (Kac+2), a positively charged residue at (Kac-1), and an aromatic residue at (Kac-2). PMID- 18400186 TI - Cytokines as therapeutic targets: advances and limitations. AB - Biological therapies targeting cytokines, T cells, or B cells have improved outcomes of inflammatory diseases. However, many issues remain open: What is the best target? How well can response be predicted? How can cure be achieved? PMID- 18400187 TI - Th17 cell differentiation: the long and winding road. AB - The characterization of the new lineage of IL-17-producing CD4+ T helper (Th17) cells has revolutionized our current understanding of T cell-mediated immunity. Over the past five years, there have been many twists and turns as the pathways that lead to Th17 cell differentiation have been elucidated. Not least of these was the discovery that TGF-beta is a crucial cytokine for Th17 cell development, suggesting that Th17 and regulatory T cell subsets share reciprocal developmental pathways during the pathogenesis or control of inflammation. This review aims to bring together the observations that have formed current opinion on factors that promote and contain Th17 cell development, in both mouse and man. Unresolved controversies in this field are also discussed: For example, IL-23 is absolutely required for disease pathogenesis in many models of Th17-cell-mediated autoimmunity, yet its role in Th17 cell development is relatively unclear. PMID- 18400188 TI - The biological functions of T helper 17 cell effector cytokines in inflammation. AB - T helper 17 (Th17) cells belong to a recently identified T helper subset, in addition to the traditional Th1 and Th2 subsets. These cells are characterized as preferential producers of interleukin-17A (IL-17A), IL-17F, IL-21, and IL-22. Th17 cells and their effector cytokines mediate host defensive mechanisms to various infections, especially extracellular bacteria infections, and are involved in the pathogenesis of many autoimmune diseases. The receptors for IL-17 and IL-22 are broadly expressed on various epithelial tissues. The effector cytokines of Th17 cells, therefore, mediate the crucial crosstalk between immune system and tissues, and play indispensable roles in tissue immunity. PMID- 18400189 TI - Contextual regulation of inflammation: a duet by transforming growth factor-beta and interleukin-10. AB - Transforming growth factor-beta (TGF-beta) and interleukin-10 (IL-10) are regulatory cytokines with pleiotropic roles in the immune system. The prominent function of TGF-beta is to maintain T cell tolerance to self or innocuous environmental antigens via its direct effects on the differentiation and homeostasis of effector and regulatory T cells. A critical route for the regulation of T cells by TGF-beta is via activation of a T cell-produced latent form of TGF-beta1 by dendritic cell-expressed avbeta8 integrin. IL-10 operates primarily as a feedback inhibitor of exuberant T cell responses to microbial antigens. T cells are also the principal producers of IL-10, the expression of which is regulated by IL-27, IL-6, and TGF-beta. The collective activity of TGF beta and IL-10 ensures a controlled inflammatory response specifically targeting pathogens without evoking excessive immunopathology to self-tissues. PMID- 18400191 TI - B cell development: important work for ERK. AB - The B cell receptor (BCR) activates the ERK kinases, but what does ERK do? In this issue of Immunity, Yasuda et al. (2008) show that ERK controls a transcription factor network by which the pre-BCR drives early B cell development. PMID- 18400190 TI - Cytokine signaling modules in inflammatory responses. AB - Cytokine signaling via a restricted number of Jak-Stat pathways positively and negatively regulates all cell types involved in the initiation, propagation, and resolution of inflammation. Here, we focus on Jak-Stat signaling in three major cell types involved in inflammatory responses: T cells, neutrophils, and macrophages. We summarize how the Jak-Stat pathways in these cells are negatively regulated by the Suppressor of cytokine signaling (Socs) proteins. We emphasize that common Jak-Stat-Socs signaling modules can have diverse developmental, pro- and anti-inflammatory outcomes depending on the cytokine receptor activated and which genes are accessible at a given time in a cell's life. Because multiple components of Jak-Stat-Socs pathways are mutated or closely associated with human inflammatory diseases, and cytokine-based therapies are increasingly deployed to treat inflammation, understanding cytokine signaling will continue to advance our ability to manipulate chronic and acute inflammatory diseases. PMID- 18400192 TI - The STATs on dendritic cell development. AB - Dendritic cells are a heterogeneous group of antigen presenting cells. In this issue of Immunity, Esashi et al. (2008) demonstrate how cytokine-receptor regulated downstream transcription factors direct dendritic cell subpopulation differentiation from hematopoietic progenitor cells. PMID- 18400193 TI - Cancer immunosurveillance: NKG2D breaks cover. AB - The idea of cancer immunosurveillance has regained interest in recent years. In this issue of Immunity, Guerra et al. (2008) present the first genetic evidence for surveillance of primary tumors by a natural killer cell-associated receptor. PMID- 18400194 TI - Basophils are back! AB - In this issue of Immunity, Tsujimura et al. (2008) report that the release of platelet-activating factor by basophils stimulated with immunoglobulin G1 (IgG1) antigen immune complexes contributes substantially to the expression of an IgG1 dependent alternative pathway of systemic anaphylaxis in mice. PMID- 18400196 TI - NO detection in biological samples: differentiation of 14 NO and 15 NO using infrared laser spectroscopy. AB - Accurate characterization of the biochemical pathways of nitric oxide (NO) is essential for investigations in the field of NO research. To analyze the different reaction pathways of enzymatic and non-enzymatic NO formation, determination of the source of NO is crucial. Measuring NO-related products in biological samples distinguishing between (14)NO and (15)NO offers the opportunity to specifically analyze NO signaling in blood and tissue. The aim of this study was to establish a highly sensitive technique for the specific measurement of NO in an isotopologue-selective manner in biological samples. With the cavity leak-out spectroscopy setup (CALOS) a differentiation between (14)NO and (15)NO is feasible. We describe here the employment of this method for measurements in biological samples. Certified gas mixtures of (14)NO/N(2) and (15)NO/N(2) were used to calibrate the system. (14)NO2- and (15)NO2- of aqueous and biological samples were reduced in a triiodide solution, and the NO released was detected via CALOS. Gas-phase chemiluminescence detection (CLD) was used for evaluation. The correlation received for both methods for the detection of NO in the gas phase was r=0.999, p<0.0001. Results obtained using aqueous and biological samples verified that CALOS enables NO measurements with high accuracy (detection limit for (14)NO2- 0.3 pmol and (15)NO2- 0.5 pmol; correlation (14)NO: p<0.0001, r=0.975, (15)NO: p<0.0001, r=0.969). The CALOS assay represents an extension of NO measurements in biological samples, allowing specific investigations of enzymatic and non-enzymatic NO formation and metabolism in a variety of samples. PMID- 18400195 TI - Interleukin-23 restrains regulatory T cell activity to drive T cell-dependent colitis. AB - Interleukin-23 (IL-23) is an inflammatory cytokine that plays a key role in the pathogenesis of several autoimmune and inflammatory diseases. It orchestrates innate and T cell-mediated inflammatory pathways and can promote T helper 17 (Th17) cell responses. Utilizing a T cell transfer model, we showed that IL-23 dependent colitis did not require IL-17 secretion by T cells. Furthermore, IL-23 independent intestinal inflammation could develop if immunosuppressive pathways were reduced. The frequency of naive T cell-derived Foxp3+ cells in the colon increased in the absence of IL-23, indicating a role for IL-23 in controlling regulatory T cell induction. Foxp3-deficient T cells induced colitis when transferred into recipients lacking IL-23p19, showing that IL-23 was not essential for intestinal inflammation in the absence of Foxp3. Taken together, our data indicate that overriding immunosuppressive pathways is an important function of IL-23 in the intestine and could influence not only Th17 cell activity but also other types of immune responses. PMID- 18400197 TI - Suture-assisted vs forceps-assisted insertion of the donor lenticula during Descemet stripping automated endothelial keratoplasty. AB - PURPOSE: To compare a new insertion method using a suture with the standard forceps-assisted insertion of the lamellar corneal graft in Descemet stripping automated endothelial keratoplasty (DSAEK). DESIGN: Interventional, comparative case series. METHODS: DSAEK was performed on 28 consecutive eyes of 28 patients (mean age +/- standard deviation, 72.5 +/- 9.7 years; 12 males). The donor tissue was prepared using the Moria automated lamellar therapeutic keratectomy microkeratome head with a 300-mm blade depth on a Moria artificial anterior chamber (Moria, Antony, France). The central 9 mm of the recipient's Descemet membrane were stripped through a superior limbal incision. In eight consecutive unselected patients, a 10-0 monofilament suture on a long straight needle was used to insert the folded donor lenticula into the recipient's eye, whereas in the other 20 patients, a standard insertion forceps was used. During the suture insertion, an anterior chamber maintainer was used to fill the anterior chamber continuously with balanced salt solution to prevent its collapse. RESULTS: Suture assisted DSAEK did not differ significantly from forceps-assisted DSAEK in any of the preoperative parameters. No significant differences were noted in the intraoperative or postoperative complications. Six months after surgery, there were no significant differences in endothelial cell counts (1804 vs 1935 cells/mm(2), respectively), visual acuity (20/40 vs 20/36, respectively), or refraction (spherical equivalent, 0.93 vs 1.05 diopters, respectively). CONCLUSIONS: Suture-assisted DSAEK is a valid alternative to forceps insertion; however, it does not seem to offer a significant advantage. PMID- 18400198 TI - Photodynamic therapy for predominantly hemorrhagic lesions in neovascular age related macular degeneration. AB - PURPOSE: To evaluate the response of eyes treated with photodynamic therapy (PDT) for predominantly hemorrhagic lesions in neovascular age-related macular degeneration (NVAMD). DESIGN: Consecutive, interventional case series. METHODS: A retrospective review was conducted of eyes with NVAMD that were treated with PDT for predominantly hemorrhagic subfoveal lesions. Baseline data recorded were patient age, use of aspirin or warfarin, history of previous PDT treatment, visual acuity (VA), extent of subretinal hemorrhage (SRH), and central foveal thickness (CFT) and total macular volume (TMV) measured by optical coherence tomography (OCT). Follow-up VA, SRH status, and complications were recorded. RESULTS: Twenty-one eyes of 21 patients were identified with a mean posttreatment follow-up of 9.4 months. Mean pretreatment VA was 20/210, and mean SRH size was 16 disk areas. The SRH resolved in 18 eyes (86%) after a mean of six months, with a mean VA of 20/210. Mean VA for all eyes at most recent follow-up was 20/250. Thirteen (62%) of 21 eyes exhibited stable or improved VA at the most recent follow-up. VA one year after PDT inversely correlated with patient age (P = .045) and initial SRH size (P = .04) and positively correlated with pretreatment VA (P = .01). Previous PDT treatment, use of aspirin or warfarin, and OCT data did not significantly correlate with VA or SRH resolution. CONCLUSIONS: PDT may be an effective treatment for minimizing visual loss in eyes with NVAMD and predominantly hemorrhagic lesions. Further study is warranted to detail the risk benefit ratio and compare PDT with other treatment modalities. PMID- 18400199 TI - Association of LOC387715 A69S with vitreous hemorrhage in polypoidal choroidal vasculopathy. AB - PURPOSE: To investigate whether the LOC387715 polymorphism is associated with polypoidal choroidal vasculopathy (PCV) and with vitreous hemorrhage (VH), one of the most severe clinical phenotypes, in the Japanese population. DESIGN: Cross sectional case-control association study. METHODS: One hundred and nine Japanese patients with PCV, composed of nine patients associated with VH (VH group) and 100 patients without VH (non-VH group), and 85 control subjects were analyzed for the LOC387715 polymorphism (rs = 10490924), using denaturing high-performance chromatography. RESULTS: There was a significant difference in the T allele frequency between PCV patients and control subjects (P < .0001). In comparison with wild-type homozygosity (GG), homozygosity for the at-risk allele genotype (TT) increased the likelihood for PCV 8.4-fold (3.6 to 19.5, 95% confidence interval [CI]) and heterozygosity for the at-risk allele genotype (TG) increased the likelihood for PCV 4.0-fold (1.9 to 8.4, 95% CI). There was a significant difference in the genotypic frequency at the LOC387715 site between the VH and non-VH groups (P = .0099, Chi-square test) with the TT genotype occurring in 88.9% in the VH group and 37.0% in the non-VH group. The frequency of the T allele in the VH group was significantly greater than that in the non-VH group (0.944 vs 0.610; P = .0039, Fisher exact test). CONCLUSIONS: The LOC387715 polymorphism is associated with PCV and clinical severity in the subgroups of PCV in the Japanese population. PMID- 18400200 TI - Sneezing reflex associated with intravenous sedation and periocular anesthetic injection. AB - PURPOSE: To assess and compare the frequency of reflex sneezing occurring during periocular anesthetic injections with and without intravenous sedation. DESIGN: Retrospective, comparative case series. METHODS: Seven hundred and twenty-two patients undergoing oculoplastic surgical procedures were included in this study. Those who received a periocular anesthetic injection under intravenous sedation served as the test group of 381 subjects. Those who received a periocular anesthetic injection without intravenous sedation served as the control group of 341 subjects. The absence or presence of reflex sneezing in both groups was recorded and compared using Chi-square analysis. RESULTS: Of the 381 patients who received periocular anesthetic injections under intravenous sedation, 19 (5%) exhibited a vigorous sneeze. Conversely, none of the 341 patients who received periocular anesthetic injections without intravenous sedation sneezed (P < or = .001). CONCLUSIONS: Surgeons, anesthesia staff, and other operating room personnel should be aware of this unusual and potentially dangerous sneeze phenomenon when periocular anesthetic injections are delivered under intravenous sedation to reduce potential ocular complications. PMID- 18400201 TI - Vision-related quality of life and visual function after retinal detachment surgery. AB - PURPOSE: To compare the vision-related quality of life (VR QOL) after rhegmatogenous retinal detachment (RD) surgery with that of normal controls and to investigate the relationship between the VR QOL and visual function after surgery for RD. DESIGN: Prospective, consecutive, comparative case series. METHODS: The 25-Item National Eye Institute Visual Function Questionnaire (VFQ 25) was self-administered by 51 RD patients at six months after surgery. Among the patients with RD, 33 underwent pars plana vitrectomy and 18 received scleral buckling. We examined the logarithm of the minimum angle of resolution (logMAR) best-corrected visual acuity (BCVA), contrast sensitivity with the CSV-1000E (Vector Vision Co, Greenville, Ohio, USA), and low-contrast visual acuity (VA) with the CSV-1000LanC10% (Vector Vision Co). From the data obtained by CSV-1000E, the area under the log contrast sensitivity function (AULCSF) was calculated. The VFQ-25 also was administered to the 46 age-matched normal controls. RESULTS: The VFQ-25 composite score and the subscales associated with near activities, mental health, dependency, and peripheral vision were significantly lower in the RD group than in the normal controls (P < .05). The VFQ-25 composite score significantly correlated with AULCSF (r = 0.354; P < .05) and low-contrast VA (r = -0.475; P < 0.001), whereas there was no correlation between the VFQ-25 composite score and logMAR BCVA (r = 0.172; P = .229). CONCLUSIONS: The VR QOL is significantly impaired in patients after surgery for RD. The disturbance in VR QOL was significantly associated with deterioration of postoperative contrast sensitivity. PMID- 18400202 TI - In vivo laser confocal microscopy after descemet stripping with automated endothelial keratoplasty. AB - PURPOSE: To investigate in vivo corneal changes in patients with bullous keratopathies before and after Descemet stripping with automated endothelial keratoplasty (DSAEK) using laser confocal microscopy. DESIGN: Single-center, prospective, comparative clinical study. METHODS: Seven patients (two men, five women; mean age, 68.9 years; range, 59 to 78) with bullous keratopathies who underwent DSAEK enrolled in this study. Laser confocal microscopy was performed before and one, three, and six months after DSAEK. Selected images were evaluated qualitatively and quantitatively for degree of haze and density of deposits. RESULTS: Preoperatively, corneal epithelial edema, subepithelial haze, keratocytes in a honeycomb pattern, and tiny needle-shaped materials in the stroma were observed in all patients. After DSAEK, subepithelial haze, donor recipient interface haze, and interface particles were observed in all five measurable cases; postoperative haze and particles decreased statistically significantly (P < .05) over follow-up. CONCLUSIONS: In vivo laser confocal microscopy is capable of identifying subclinical corneal abnormality after DSAEK with high resolution. Further studies in a large number of patients and long-term follow-up after DSAEK using this device are needed to fully understand long-term corneal stromal changes after DSAEK and whether the preexisting corneal stromal pathologies are reversible. PMID- 18400203 TI - Vision-related quality of life and visual function following vitrectomy for proliferative diabetic retinopathy. AB - PURPOSE: To evaluate the vision-related quality of life (VR-QOL) and visual function following vitrectomy for proliferative diabetic retinopathy (PDR). DESIGN: Prospective, interventional, consecutive, comparative case series. METHODS: The 25-Item National Eye Institute Visual Function Questionnaire (VFQ 25) was answered by 51 patients with PDR before and three months after 20-gauge pars plana vitrectomy. Among the patients with PDR, 30 underwent combined cataract surgery and vitrectomy. Clinical data were collected, including logarithm of minimal angle of resolution (logMAR) best-corrected visual acuity (BCVA), letter contrast sensitivity, and metamorphopsia. The VFQ-25 was also administered to 46 age-matched normal controls. RESULTS: The preoperative VFQ-25 composite score was significantly lower in the PDR patients (56.3 +/- 18.8, mean +/- standard deviation) than in the normal controls (85.2 +/- 10.3; P < .0001). Vitrectomy significantly improved VFQ-25 composite score (P < .005) and eight of 12 subscales (P < .05). The preoperative VFQ-25 composite score significantly correlated with preoperative logMAR BCVA in the better-seeing eye (P < .001). The postoperative VFQ-25 composite score exhibited significant correlation with postoperative logMAR BCVA in the worse-seeing eye (P < .0001) as well as the better-seeing eye (P < .0001). Similar significant correlations were also observed between the VFQ-25 composite score and letter contrast sensitivity. CONCLUSIONS: Vitrectomy for PDR significantly improves VR-QOL. The current study quantitatively indicates that VR-QOL in PDR patients depends on visual acuity and contrast sensitivity in the better-seeing eye preoperatively and in both the better-seeing and worse-seeing eyes postoperatively. PMID- 18400204 TI - Novel KCNV2 mutations in cone dystrophy with supernormal rod electroretinogram. AB - PURPOSE: To describe patients with cone dystrophy and supernormal rod electroretinogram (ERG) and search for mutations in the recently described KCNV2 gene. DESIGN: Clinical and molecular study. METHODS: Patients from three families originating from France, Morocco, and Algeria had standard ophthalmologic examination and color vision analysis, Goldmann perimetry, International Society for Clinical Electrophysiology of Vision (ISCEV) protocol in accordance with ERG testing, autofluorescence evaluation, and optical coherence tomography 3 scanning. The two coding exons of KCNV2 were polymerase chain reaction amplified and sequenced. RESULTS: All patients had the characteristic features of supernormal, delayed rod ERG responses at the highest levels of stimulation and markedly reduced cone responses. In the French family, two affected sisters were compound heterozygotes for the recurrent c.1381G>A (Gly461Arg) mutation and for a novel c.442G>T (Glu148Stop) mutation. In the Moroccan family, affected members were homozygotes for the novel c.1404delC mutation (His468fsX503) and in the Algerian family, the proband was homozygote for the novel c.1001delC mutation (Ala334fsX453). In the three families, parents were unaffected heterozygote carriers. None of the mutations were present in 50 control chromosomes. CONCLUSIONS: The three novel truncative mutations are likely to be null mutations leading to loss of function, with no difference in the phenotype presentation. Amino acid changes are found exclusively in the N-terminal fragment of the protein and in the P-loop, indicating the importance of those regions for the function of the KCNV2 protein. PMID- 18400205 TI - Orientation teeth in nonmechanical femtosecond laser corneal trephination for penetrating keratoplasty. AB - PURPOSE: To evaluate femtosecond laser-assisted corneal full-thickness trephination with orientation teeth and notches for penetrating keratoplasty (PKP). DESIGN: Interventional case series. METHODS: Four eyes of four patients aged 34 to 55 years underwent PKP using a femtosecond laser system enabling corneal trephination with orientation teeth and notches. Patients were affected by advanced keratoconus (n = 2) or postinfectious corneal scar (n = 2). Scanning electron microscopy (SEM) was performed to evaluate the cut quality of donor and recipient cornea. RESULTS: All procedures were completed without any complications. In all cases, the donor and the recipient cornea were cut with the same size of 8.0-mm diameter. A complete perforating cut was achieved in all eyes, and a correct intraoperative matching of teeth and notches was obtained. SEM of donor and recipient specimens showed a full-thickness cut with smooth and regular shape of orientation teeth and notches. CONCLUSIONS: Orientation teeth and notches shaped PKP using the femtosecond laser technology is feasible and may offer further promising approaches toward customized trephination techniques. PMID- 18400206 TI - Retinoids induce cytochrome P450 3A4 through RXR/VDR-mediated pathway. AB - A panel of retinoids and carotenoids was screened as potential inducers of CYP3A4 through the RXR/VDR-mediated signaling pathway. Transient transfection assays revealed that 3 out of 12 retinoids screened transactivated RXRalpha/VDR and induced CYP3A4 reporter activity. These three retinoids are the active metabolites of retinoids, 9-cis-retinal, 9-cis-retinoic acid (9-cis-RA), and all trans-retinoic acid (all-trans-RA). 9-cis-RA and all-trans-RA preferentially transactivated the RXR/VDR heterodimers and RXR homodimers. Retinoids and VDR agonist 1alpha, 25-dihydroxyvitamin D(3), but not PXR or CAR activator, could induce Cyp3a11 mRNA level in hepatocytes derived from PXR/CAR-double null mouse. Moreover, retinoids induced CYP3A4 enzyme activity in HepG2 human hepatoma and Caco-2 human colorectal adenocarcinoma cells. A direct role of retinoid-mediated CYP3A4 induction through RXRalpha/VDR was proved by the results that 9-cis retinal, 9-cis-RA, and all-trans-RA recruited RXRalpha and VDR to CYP3A4 regulatory region pER6 (proximal everted repeat with a 6-nucleotide spacer) and dXREM (distal xenobiotic-responsive enhancer module). Thus, using various approaches, we have unequivocally demonstrated that retinoids transactivate RXR/VDR heterodimers and RXR homodimers and induce CYP3A expression at mRNA as well as enzyme activity levels in both liver and intestinal cells. It is possible that retinoids might alter endobiotic metabolism through CYP3A4 induction in vivo. PMID- 18400209 TI - Tyrosine availability modulates potassium-induced striatal catecholamine efflux in vivo. AB - The relationship between tyrosine availability and high potassium (K+) induced dopamine (DA) and norepinephrine (NE) efflux was examined in striatum using in vivo microdialysis. High K+ (80 mM) was included in perfusate for two 30 min periods, 2.5 h apart. After the first high-K+ perfusion, a tyrosine- and phenylalanine-free mixture of large neutral amino acids (LNAA(-)) was administered (IP) to lower brain tyrosine. Tyrosine (0, 25, 50 or 100 mg/kg IP) was administered 30 min prior to the second high-K+ perfusion. The ratio of catecholamine efflux during the two perfusions (P2/P1) was compared between groups. LNAA(-) significantly lowered P2/P1 for both DA and NE. Tyrosine 25-50 mg/kg blocked the LNAA(-) effect. We conclude that catecholamine efflux during prolonged depolarization is tyrosine dependent. Analyses of LNAA levels suggest that availability of tyrosine for tyrosine hydroxylation may be modulated by competition between LNAAs within brain extracellular fluid. PMID- 18400208 TI - Gamma-band auditory steady-state responses are impaired in first episode psychosis. AB - BACKGROUND: In chronic schizophrenia and chronic bipolar disorder, gamma band (30 100 Hz) auditory steady-state electroencephalogram responses (ASSRs) are reduced in power and phase locking, likely reflecting neural circuit dysfunction. Here we examined whether gamma ASSR deficits are also present at first hospitalization for psychosis. METHODS: Subjects were 16 first episode schizophrenia patients (SZ), 16 first episode affective disorder patients (AFF) (13 with bipolar disorder), and 33 healthy control subjects (HC). Stimuli were 20-, 30-, and 40-Hz binaural click trains. The ASSR phase locking and evoked power were analyzed with the Morlet wavelet transform. RESULTS: At 40-Hz stimulation, SZ and AFF had significantly reduced phase locking compared with HC. This deficit was more pronounced over the left hemisphere in SZ. Evoked power at 40 Hz was also reduced in the patients compared with HC. At 30-Hz stimulation phase locking and evoked power were reduced in both patient groups. The 20-Hz ASSR did not differ between groups, but phase locking and evoked power of the 40-Hz harmonic of the 20-Hz ASSR were reduced in both SZ and AFF. Phase locking of this 40-Hz harmonic was correlated with total positive symptoms in SZ. CONCLUSIONS: The gamma ASSR deficit is present at first hospitalization for both schizophrenia and affective disorder but shows a left hemisphere bias in first hospitalized SZ. Some of the neural circuitry abnormalities underlying the gamma ASSR deficit might be common to psychoses in general, whereas others might be specific to particular disorders. PMID- 18400207 TI - The cognitive neuroscience of working memory: relevance to CNTRICS and schizophrenia. AB - Working memory is one of the central constructs in cognitive science and has received enormous attention in the theoretical and empirical literature. Similarly, working memory deficits have long been thought to be among the core cognitive deficits in schizophrenia, making it a ripe area for translation. This article provides a brief overview of the current theories and data on the psychological and neural mechanisms involved in working memory, which is a summary of the presentation and discussion on working memory that occurred at the first Cognitive Neuroscience Treatment Research to Improve Cognition in Schizophrenia (CNTRICS) meeting (Washington, D.C.). At this meeting, the consensus was that the constructs of goal maintenance and interference control were the most ready to be pursued as part of a translational cognitive neuroscience effort at future CNTRICS meetings. The constructs of long-term memory reactivation, capacity, and strategic encoding were felt to be of great clinical interest but requiring more basic research. In addition, the group felt that the constructs of maintenance over time and updating in working memory had growing construct validity at the psychological and neural levels but required more research in schizophrenia before these should be considered as targets for a clinical trials setting. PMID- 18400210 TI - Activation of 5-HT2C receptors acutely induces Per1 gene expression in the rat SCN in vitro. AB - The suprachiasmatic nucleus of the hypothalamus receives dense serotonergic projections from the raphe nuclei. This input may be involved in the transmission of photic information with activation of 5-HT receptors at night having a photo mimetic effect. 5-HT2C receptors have been implicated in this role, with 5-HT2C receptor agonist administration at night resulting in the induction of cFOS protein, and a phase delay of melatonin metabolite excretion rhythms in a manner similar to light. To investigate the site of action of 5-HT2C receptor agonists, rat SCN explants were isolated in culture, allowing exposure of agonists to denervated tissue. Any response could then be attributed to the actions of the 5 HT2C receptor agonist on cells within the SCN, rather than at other distant sites or on pre-synaptic receptors. Rat SCN tissue was microdissected and prepared in tissue culture conditions. The acute effect of the 5-HT2A/2C receptor agonist DOI and glutamate administration at various circadian times on c-fos and Per1 expressions was assessed. Glutamate induced the expression of c-fos at CT6, CT16 and CT22, but induced Per1 expression at CT16 only. DOI altered c-fos expression at all times examined with an increase at CT6 and CT22, and a repression at CT16. However, DOI significantly increased Per1 expression when applied at CT16, but had no effect at either CT22 or CT6. This response was similar to that previously observed in vivo. This provides support for the hypothesis that the actions of 5 HT2C receptor agonists are on cells located within the SCN. PMID- 18400211 TI - Ferulic acid provides neuroprotection against oxidative stress-related apoptosis after cerebral ischemia/reperfusion injury by inhibiting ICAM-1 mRNA expression in rats. AB - Our previous studies have shown that ferulic acid (4-hydroxy-3-methoxycinnamic acid, FA) inhibits intercellular adhesion molecule-1 (ICAM-1) expression in the ischemic striatum after 2 h of reperfusion in a transient middle cerebral artery occlusion model in rats. The purpose of this study is to further investigate the neuroprotective effects of FA during reperfusion after cerebral ischemia. Rats were subjected to 90 min of ischemia; they were then sacrificed after 2, 10, 24 and 36 h of reperfusion. ICAM-1 and macrophage-1 antigen (Mac-1) mRNA were detected using semi-quantitative RT-PCR at 2 h of reperfusion. Mac-1, 4-hydroxy-2 nonenal (4-HNE), 8-hydroxy-2'-deoxyguanosine (8-OHdG), active caspase 3, neuronal nuclei (NeuN) and TUNEL positive cells were measured at 2, 10, 24 and 36 h of reperfusion. FA (100 mg/kg, i.v.) administered immediately after MCAo inhibited ICAM-1 and Mac-1 mRNA expression in the striatum at 2 h of reperfusion, and reduced the number of Mac-1, 4-HNE and 8-OHdG positive cells in the ischemic rim and core at 10, 24 and 36 h of reperfusion. FA decreased TUNEL positive cells in the penumbra at 10 h, and in the ischemic boundary and core at 24 and 36 h of reperfusion. FA curtailed active caspase 3 expression in the penumbra at 10 h and restored NeuN-labeled neurons in the penumbra and ischemic core at 36 h of reperfusion. FA decreased the level of ICAM-1 mRNA and the number of microglia/macrophages, and subsequently down-regulated inflammation-induced oxidative stress and oxidative stress-related apoptosis, suggesting that FA provides neuroprotection against oxidative stress-related apoptosis by inhibiting ICAM-1 mRNA expression after cerebral ischemia/reperfusion injury in rats. PMID- 18400213 TI - TTX-R Na+ current-reduction by celecoxib correlates with changes in PGE(2) and CGRP within rat DRG neurons during acute incisional pain. AB - The present study was undertaken to investigate whether celecoxib could regulate the tetrodotoxin-resistant (TTX-R) sodium channel current in rat dorsal root ganglia (DRG) and whether prostaglandin E2 (PGE2) and calcitonin gene-related protein (CGRP) were involved in celecoxib's analgesia during acute incisional pain. Seventy-five rats were randomly allocated into three groups. Group A was the control group receiving a placebo (sugar pill) 1 h before and 12 h after surgery (right hind paw incisional pain). Group B was the test group receiving celecoxib 30 mg/kg orally 1 h before and 12 h after surgery. Group C was the naive group receiving a sham operation. The changes in the mechanical withdrawal thresholds, PGE2 and CGRP concentration in incisional paw tissue and DRG, and total TTX-R sodium channel current density in small DRG neurons were investigated 1 h before the operation and 2 h, 6 h, 12 h, 24 h, 48 h and 96 h after the operation. The results showed both of a decrease in mechanical withdrawal thresholds and an increase of TTX-R sodium channel current density in DRG neurons in group B were significantly lower than those of group A at 24 h and 48 h after the operation (P<0.05). The increase in PGE2 and CGRP concentrations at incisional paw tissue and DRG neurons in group B were lower than those of groups A at 24 h and 48 h after the operation (P<0.05). This study indicates that: 1) celecoxib can inhibit TTX-R sodium channel current density in rat DRG neurons; 2) PGE2 and CGRP participate in celecoxib's analgesic effect on acute incisional pain. PMID- 18400212 TI - Losartan improves impaired nitric oxide synthase-dependent dilatation of cerebral arterioles in type 1 diabetic rats. AB - We examined whether activation of angiotensin-1 receptors (AT1R) could account for impaired responses of cerebral arterioles during type 1 diabetes (T1D). First, we measured responses of cerebral arterioles in nondiabetic rats to eNOS dependent (acetylcholine and adenosine diphosphate (ADP)) and -independent (nitroglycerin) agonists before and during application of angiotensin II. Next, we examined whether losartan could improve impaired responses of cerebral arterioles during T1D. In addition, we harvested cerebral microvessels for Western blot analysis of AT1R protein and measured production of superoxide anion by brain tissue under basal conditions and in response to angiotensin II in the absence or presence of losartan. We found that angiotensin II specifically impaired eNOS-dependent reactivity of cerebral arterioles. In addition, while losartan did not alter responses in nondiabetics, losartan restored impaired eNOS dependent vasodilatation in diabetics. Further, AT1R protein was higher in diabetics compared to nondiabetics. Finally, superoxide production was higher in brain tissue from diabetics compared to nondiabetics under basal conditions, angiotensin II increased superoxide production in nondiabetics and diabetics, and losartan decreased basal (diabetics) and angiotensin II-induced production of superoxide (nondiabetics and diabetics). We suggest that activation of AT1R during T1D plays a critical role in impaired eNOS-dependent dilatation of cerebral arterioles. PMID- 18400214 TI - Amyloid peptides in different assembly states and related effects on isolated and cellular proteasomes. AB - The role of amyloid-beta protein (Abeta) in the pathogenesis of Alzheimer's disease (AD) has been widely investigated and amyloid aggregates are considered a major cause of neuronal dysfunction. Increasing evidence has identified a correlation between this protein and the proteasome, the cellular proteolytic machinery, in particular the ubiquitin-proteasome system. The 20S proteasome is the catalytic core of a complex, known as 26S proteasome, and is the main responsible for the clearance of misfolded and oxidized proteins. In this work we have investigated the effects of different assembly states of two major amyloid peptides, Abeta (1-40) and Abeta (1-42) on the 20S proteasome functionality and on the ubiquitin-dependent pathway of protein degradation. In particular, we have tested proteasome activities after Abeta treatment on purified 20S complexes and on lysates of a human neuroblastoma cell line. Our findings show a significant decrease in proteasome activity, more evident in cell lysates than in isolated complexes, and an increased amount of ubiquitin-protein conjugates and of a known proteasome substrate (p27). Furthermore, the altered proteasome functionality is not associated with a decrease in cell viability, but is linked with increased levels of protein oxidation. PMID- 18400215 TI - Distribution of somatostatin immunoreactive neurons and fibres in the central nervous system of a chondrostean, the Siberian sturgeon (Acipenser baeri). AB - Somatostatin (SOM) is a neuropeptide that is widely distributed in the central nervous system of vertebrates. Two isoforms of somatostatin (SS1 and SS2) have been characterized in sturgeon and in situ hybridisation studies in the sturgeon brain have demonstrated that mRNAs of the two somatostatin precursors (PSS1 and PSS2) are differentially expressed in neurons [Trabucchi, M., Tostivint, H., Lihrmann, I., Sollars, C., Vallarino, M., Dores, R.M., Vaudry, H., 2002. Polygenic expression of somatostatin in the sturgeon Acipenser transmontanus: molecular cloning and distribution of the mRNAs encoding two somatostatin precursors. J. Comp. Neurol. 443, 332-345.]. However, neither the morphology of somatostatinergic neurons nor the patterns of innervation have yet been characterized. To gain further insight into the evolution of this system in primitive bony fishes, we studied the distribution of somatostatin-immunoreactive (SOM-ir) cells and fibres in the brain of the Siberian sturgeon (Acipenser baeri). Most SOM-ir cells were found in the preoptic area and hypothalamus and abundant SOM-ir fibres coursed along the hypothalamic floor towards the median eminence, suggesting a hypophysiotrophic role for SOM in sturgeon. In addition, SOM-ir cells and fibres were observed in extrahypothalamic regions such as the telencephalon thalamus, rhombencephalon and spinal cord, which also suggests neuromodulatory and/or neurotransmitter functions for this peptide. Overall there was a good correlation between the distribution of SOM-ir neurons throughout the brain of A. baeri and that of PSS1 mRNA in Acipenser transmontanus. Comparative analysis of the results with those obtained in other groups of fishes and tetrapods indicates that widespread distribution of this peptide in the brain is shared by early vertebrate lines and that the general organization of the somatostatinergic systems has been well-conserved during evolution. PMID- 18400216 TI - Potential use of hirudin in diabetic cataract: a study of galactose mediated human lens epithelial cells injury. AB - Osmotic stress, together with weakened antioxidant defense mechanisms, is attributed to the changes observed in human diabetic cataract. The use of hirudin, an antithrombic agent, in the pathogenesis of human cataracts has not been studied so far. Since the epithelium is the metabolic unit of the lens, the effect of recombinant hirudin variant III (rHV3) on galactose-induced morphological changes and antioxidant status of human lens epithelial line SRA01/04 in culture was evaluated in this study. The human lens epithelial cells (hLECs) were cultured in D/F(12) medium (normal group), D/F(12) medium + 50 mM D galactose (control group) or D/F(12) medium + 50 mM D-galactose+rHV3 (test group) for 24 or 72 h. The cells were observed under the light, fluorescence and transmission electron microscope for any morphological changes, while the cell viability was assessed by methylthiazol tetrazolium (MTT) assay. The cells in flasks were harvested for the estimation of various antioxidant parameters. Cell morphology, viability, malondialdeyde, glutathione and antioxidant enzymes were significantly altered in the control group as compared with the normal group. Administration of rHV3 confers significant protection against these changes in the human lens epithelial cells. These results demonstrated that rHV3 could effectively protect galactose-induced hLEC injury and suggested that it could have potential use in diabetic cataracts. PMID- 18400217 TI - On the role of regular phonological variation in lexical access: evidence from voice assimilation in French. AB - The present study investigated whether lexical access is affected by a regular phonological variation in connected speech: voice assimilation in French. Two associative priming experiments were conducted to determine whether strongly assimilated, potentially ambiguous word forms activate the conceptual representation of the underlying word. Would the ambiguous word form [sud] (either assimilated soute 'hold' or soude 'soda') facilitate "bagage" 'luggage', which is semantically related to soute but not to soude? In Experiment 1, words in either canonical or strongly assimilated form were presented as primes. Both forms primed their related target to the same extent. Potential lexical ambiguity did not modulate priming effects. In Experiment 2, the primes such as assimilated soute pronounced [sud] used in Experiment 1 were replaced with primes such as soude canonically pronounced [sud]. No semantic priming effect was obtained with these primes. Therefore, the effect observed for assimilated forms in Experiment 1 cannot be due to overall phonological proximity between canonical and assimilated forms. We propose that listeners must recover the intended words behind the assimilated forms through the exploitation of the remaining traces of the underlying form, however subtle these traces may be. PMID- 18400218 TI - Continuous theta-burst stimulation over the dorsal premotor cortex interferes with associative learning during object lifting. AB - When lifting objects of different mass, humans scale grip force according to the expected mass. In this context, humans are able to associate a sensory cue, such as a colour, to a particular mass of an object and link this association to the grip forces necessary for lifting. Here, we study the role of the dorsal premotor cortex (PMd) in setting-up an association between a colour cue and a particular mass to be lifted. Healthy right-handed subjects used a precision grip between the index finger and thumb to lift two different masses. Colour cues provided information about which of the two masses subjects would have to lift. Subjects first performed a series of lifts with the right hand to establish a stable association between a colour cue and a mass, followed by 20sec of continuous high frequency repetitive trancranial magnetic stimulation using a recently developed protocol (continuous theta-burst stimulation, cTBS) over (i) the left primary motor cortex, (ii) the left PMd and (iii) the left occipital cortex to be commenced by another series of lifts with either the right or left hand. cTBS over the PMd, but not over the primary motor cortex or O1, disrupted the predictive scaling of isometric finger forces based on colour cues, irrespective of whether the right or left hand performed the lifts after the stimulation. Our data highlight the role of the PMd to generalize and maintain associative memory processes relevant for predictive control of grip forces during object manipulation. PMID- 18400220 TI - Locomotor effects of morphine or alcohol in mice after a repeated treatment with the cannabinoid agonist HU 210. AB - The consequences of the consumption of cannabinoids with other drugs of abuse are of particular medical relevance. Several studies investigated the ability of cannabinoids to induce a locomotor cross-sensitization to other addictive drugs, but results remain inconsistent. Therefore, we investigated in mice the consequences of a repeated treatment with the cannabinoid agonist HU 210 on motor effects of morphine or alcohol. In mice receiving a daily injection of HU 210 (12.5 to 200 microg/kg) during 7 days, no hetero-sensitization to the stimulation induced by either morphine (7.5 mg/kg) or alcohol (1 or 1.5 g/kg) emerged, from 1 day up to 35 days after the end of the sub-chronic treatment with HU 210. Even a chronic treatment with a high dose of HU 210 (14 days, 200 microg/kg) induced no subsequent enhancement of the stimulant effects of morphine or alcohol. In fact, the motor stimulant effect of morphine or alcohol in chronically HU 210 pre treated mice was even abolished until the 3rd day of abstinence. This reduction was presumably due to residual HU 210 since this effect was prevented by the cannabinoid antagonist rimonabant. Afterwards, chronically cannabinoid pre treated mice remained less active than vehicle pre-treated mice from the 7th day up to the 35th day after the end of the 14-day treatment with HU 210. In conclusion, we failed to detect any hetero-sensitization whatever the pre treatment regimen. However, only after the 14-day regimen, HU 210 pre-treated mice displayed a long-lasting decrease in activity, suggesting that some neuronal adaptive changes may have occurred. PMID- 18400219 TI - Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. AB - Despite significant advances in identifying signaling molecules that induce cardiogenesis in mammals, the transcription factors that control the onset of cardiac myocyte gene expression have remained elusive. Candidates include the zinc finger transcription factors GATA binding proteins 4 and 6 (GATA4, GATA6). The individual loss of either protein in mice results in lethality prior to the onset of heart development due to defects in the extra-embryonic endoderm; however, when this extra-embryonic deficiency is circumvented using tetraploid embryo complementation, cardiac myocyte differentiation initiates normally. Here we show that these factors have redundant roles in controlling the onset of cardiac myocyte differentiation. As a consequence, Gata4(-/-)Gata6(-/-) embryos completely lack hearts, although second heart field progenitor cells are still generated. Our data support a model whereby GATA4 or GATA6 are essential for expression of the network of transcription factors that regulate the onset of cardiac myocyte gene expression during mammalian development. PMID- 18400221 TI - Moving towards therapies for juvenile Batten disease? AB - The neuronal ceroid lipofuscinoses (NCLs or Batten disease) are a group of at least nine autosomal recessively inherited monogenetic storage disorders. Because there are no effective therapies available, all forms of NCL invariably prove fatal after a prolonged period of disability. Indeed, for the forms of NCL that are the result of mutations in transmembrane proteins, the therapeutic outlook remains uniformly bleak. This includes juvenile NCL (JNCL); the most prevalent form of Batten disease that is the result of mutations in the CLN3 gene. Characterizing Cln3 deficient mice is now revealing important clues about the pathogenesis of JNCL. This includes evidence for elevated levels of glutamate within the JNCL CNS and cell type selective sensitivity to alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionate (AMPA) type glutamate receptor overactivation. These findings raise the possibility that AMPA-receptor blockade may potentially be beneficial in JNCL. This possibility has now begun to be tested in Cln3 mutant mice using a single intraperitoneal injection of the non-competitive AMPA antagonist EGIS-8332 by Kovacs and Pearce [Kovacs, A.D., Pearce, D.A., 2008. Attenuation of AMPA receptor activity improves motor skills in a mouse model of juvenile Batten disease. Exp. Neurol. 209, 288-291.]. Although a positive effect of upon motor coordination deficits in this mouse model of JNCL is reported in this acute study, it remains unclear whether EGIS-8332 provides any lasting benefit or effects upon other aspects of their disease phenotype. Although supplying the first evidence for any form of improvement in a disease-relevant phenotype in Cln3 mutant mice, more detailed studies will be needed to determine whether these preliminary findings will translate into a successful therapy for either murine or human JNCL. PMID- 18400222 TI - GnRH immunodetection in the brain of the holocephalan fish Chimaera monstrosa L.: correlation to oocyte maturation. AB - Chimera monstrosa (rabbit fish) like other holocephalans is a rare, delicate deep sea fish. Owing to the difficulty of sampling individuals in good shape, there is a paucity of information available on the morphology and physiology of this species especially concerning reproduction. In holocephalans, a hypothalamus pituitary-gonadal axis has been postulated and a GnRH molecule identical to cGnRH II has been identified. The aim of the present study was to correlate the presence of steroidogenic enzymes in the ovarian follicles with the presence of GnRH in the hypothalamus. Estrogens, the steroids that trigger the accumulation of yolk (vitellogenesis) in the oocytes are synthesized by the somatic cells of the follicle in the vitellogenic stages via a cascade of steroid dehydrogenases involving 3 beta-hydroxysteroid-dehydrogenase (3 beta-HSD; in the inner thecal layer) and aromatase cytochrome (P450; granulosa layer). Our results showed that 3 beta-HSD is present concomitant with the presence of cGnRH II in the preoptic area and in the ventral hypothalamus. Another form of immunoreactive GnRH, mGnRH is also present in the brain of C. monstrosa. It is localized in the ventral telencephalon and in the midbrain caudal diencephalon (boundary between ventral thalamus and tegmentum of the mesencephalon). This form of GnRH is probably correlated with sexual behaviour. PMID- 18400223 TI - Identification of unique thyrotropin receptor (TSHR) splice variants in the chicken: the chicken TSHR gene revisited. AB - We previously described the cloning of the full-length chicken thyrotropin receptor (TSHRa) and two splice variants, lacking exon 3 (TSHRb) or both exons 2 and 3 (TSHRc). Here we report the identification of three novel splice variants of the chicken TSHR, named TSHRd, -e and -f, differing in their C-terminal region and containing unique exonic sequences that are not present in the other TSHR variants. This finding suggests a TSHR gene structure with 13 rather than the previously assumed 10 exons. The three novel exons appear to be chicken-specific, as no equivalents of these exons were found in other vertebrate genomes. Like the full-length receptor, the five TSHR splice variants are most abundantly expressed in thyroid gland. TSHRb, -d, -e and -f mRNA was also present in virtually all extra-thyroidal tissues expressing TSHRa, whereas TSHRc shows a more restricted tissue distribution. Whether these receptor transcripts are translated to functional proteins needs to be verified, but if so, they could be attributed various physiological roles. PMID- 18400224 TI - Peptidomic approach based on combined capillary isoelectric focusing and mass spectrometry for the characterization of the plasmin primary products from bovine and water buffalo beta-casein. AB - The main peptides produced by hydrolysis of water buffalo beta-casein with plasmin were characterized by capillary electrophoresis and mass spectrometry and compared with their bovine homologous. A novel breakdown product arising from the hydrolysis of water buffalo beta-casein, originated by the presence of a plasmin sensitive Lys bond at position 68 was identified, which was not present in bovine beta-casein. On the basis of this evidence, an improved procedure for the detection and the differentiation of the products of plasmin hydrolysis of bovine and water buffalo beta-casein by capillary isoelectric focusing was set-up. In the experimental conditions, the gamma-casein from the two species was efficiently separated. Comparison of the capillary electropherograms with those obtained by ultra-thin-layer isoelectric focusing, the reference method for routine analysis of plasmin digests of casein, suggests that capillary electrophoresis isoelectric focusing may constitute a successful alternative to the traditional slab gel electrophoresis analysis of plasmin digests of casein either for basic structural studies or for applications in the quality assessment of dairy products. PMID- 18400225 TI - Isolation of Indonesian cananga oil using multi-cycle pressure drop process. AB - New process, instantaneous controlled pressure drop (DIC) was applied on Cananga odorata dry flowers with the aim to isolate essential oil. DIC is based on high temperature, short time heating followed by an abrupt pressure drop into a vacuum. A part of volatile compounds is carried away from flowers in the form of vapor (DIC direct oil) that evolves adiabatically during the pressure drop (proper isolation process) and the other part remains in the DIC-treated flowers (DIC residual oil). In the present paper, the effect of DIC cycle number (1-9) and heating time (4.3-15.7 min) on the availability of oil compounds was investigated at three levels of steam pressure (0.28, 0.4 and 0.6 MPa). The availability was defined as the amount of a compound in direct or residual oil divided by the amount of this compound in the reference oil extracted from non treated flowers by chloroform during 2h. The total availability and yield of volatiles in the direct oil increased with pressure and cycle number. At a higher pressure, the effect of heating time was insignificant. The amount of oxygenated monoterpenes and other light oxygenated compounds (i.e. predominantly exogenous compounds) in the residual flowers was lower than in the direct oil and this amount decreased with cycle number. On the other hand, the availability of oxygenated sesquiterpenes and other heavy oxygenated compounds (i.e. predominantly endogenous compounds) in residual flowers exhibited a maximum for about five cycles and their quantity at this point was three times as much as in the direct oil. The total availability of each compound at 0.6 MPa was higher than one. The rapid DIC process (0.6 MPa, 8 cycles, 6 min) gave better results than steam distillation (16 h) concerning direct oil yield (2.8%dm versus 2.5%dm) and content of oxygenated compounds (72.5% versus 61.7%). PMID- 18400226 TI - Chromatographic characterisation of monolithic capillary columns for liquid chromatography based on methyltrimethoxysilane as sole precursor. AB - A set of monolithic capillary columns for liquid chromatography, synthesized according to a recently developed protocol which uses methyltrimethoxysilane (MTMS) as a sole precursor, is characterised by various chromatographic tests to assess its physico-chemical properties. The new stationary phase material shows a hydrophobicity (assessed on the basis of methylene selectivity) comparable to commercial C(8) columns. The MTMS-based columns exhibit a reduced affinity towards planar molecules such as PAHs, compared to C(18) modified columns, which can be explained by a retention mechanism that is more governed by adsorption rather than partitioning. In comparison to commercial products an only moderate silanol activity was observed, even without any endcapping procedure applied. Selectivity between hydrophobic test compounds showed to be uniform between the columns investigated in this study, whereas retention factors differed up to 20% (batch-to-batch reproducibility) between columns produced under the same conditions. For most of the materials investigated in this study, size exclusion towards even only slightly larger molecules such as triphenylene was observed. It was demonstrated that inclusion of a micelle-forming detergent such as Brij in the synthetic protocol could partially overcome this problem. PMID- 18400227 TI - Ultra rapid liquid chromatography as second dimension in a comprehensive two dimensional method for the screening of pharmaceutical samples in stability and stress studies. AB - Separation orthogonality has been explored with respect to comprehensive two dimensional liquid chromatography (2D-LC) for different reversed-phase stationary phases. The outcome of this study points out an SB-CN x BEH-C18 combination, used in the first and the second dimension, respectively, as the most orthogonal chromatographic system for the samples assayed. The present investigation reports the employment of an ultra high pressure liquid chromatography system (UPLC) as the second chromatographic dimension, increasing the sensitivity and the speed, completing the whole chromatographic separation in a reasonable time frame. Finally, an off-line 2D-LC method with diode array detection based on the UPLC has been optimized, allowing the separation and minimizing the run time. SB-CN and BEH-C18 were employed as first and second dimension, respectively, with gradient elution applied in each dimension. Alprazolam degraded tablets were studied as a proof of concept of the utility of this type of setups for impurity profiling of complex samples. PMID- 18400228 TI - Sample preparation optimization in wine and grapes. Dilution and sample/headspace volume equilibrium theory for headspace solid-phase microextraction. AB - Most headspace solid-phase microextraction (HS-SPME) volatile analysis methods have been developed for aqueous samples and have been either adapted or applied to complex matrices. This study examines sample/headspace equilibrium based on realistic (non-spiked) concentration levels in real complex sample matrices (grapes and wine) with a systematic multivariate statistical approach. The presence and absence of matrix effects are explained through exponential and linear relationships, respectively. The potential of over- and underestimating volatile compounds in a diluted sample is illustrated and the common dilution equation (C1V1=C2V2) is shown to not always apply to headspace volatile analysis. Additionally, sample dilution was shown to be more sensitive to matrix effects than sample/headspace volume variations with the latter showing analyte dependency. An optimum sample size of 6.9-8.6g in a 20mL vial without dilution was observed. This study shows that sensitivity and limit of detection (LOD) can be improved to a limit with a subsequent loss - an extension to existing theory. The study further illustrates that in trying to bring an analyte within linear range through sample dilution, sensitivity and LOD can be lost with a probable shift in optimum ranges and sample/headspace equilibrium. PMID- 18400229 TI - Fast analysis of volatile organic compounds and disinfection by-products in drinking water using solid-phase microextraction-gas chromatography/time-of flight mass spectrometry. AB - A fast method was developed for the extraction and analysis of volatile organic compounds, including disinfection by-products (DBPs), with headspace solid-phase microextraction (HS-SPME) and gas chromatography/mass spectrometry (GC/MS) techniques. A GC/time-of-flight (TOF)-MS instrument, which had fast acquisition rates and powerful deconvolution software, was used. Under optimum conditions total runtime was 45s. Volatile organic compounds (VOCs), including purgeable A and B compounds (listed in US Environmental Protection Agency method 624), were identified in standard water samples. Extraction times were 1min for more volatile compounds and 2min for less volatile compounds. The method was applied to the analysis of water samples treated under different disinfection processes and the results were compared with those from a liquid-liquid extraction method. PMID- 18400230 TI - Factors affecting the sorption of trivalent chromium by zeolite synthesized from coal fly ash. AB - This research was initiated to determine the effects of different constituents and properties of zeolite synthesized from fly ash (ZFA) on Cr(III) sorption. The uptake of Cr(III) by ZFA was influenced greatly by pH, increasing with the increase in pH. The pH was controlled mainly by calcium-related components (especially CaCO3 and free CaO) and zeolite components in ZFAs. Sorption maximum of Cr(III) (Qm), determined by a repeated batch equilibration method, ranged from 22.29 to 99.91 mg/g for the 14 ZFAs. The Q(m) value correlated significantly with Ca-related components. The correlation coefficients were 0.9467, 0.5469, 0.7521, and 0.9195 for total Ca, CaCO3, CaSO4, and f.CaO, respectively. The Qm value was also closely related to cation-exchange capacity (r=0.6872) and specific surface area (r=0.7249). Correlation coefficients of Qm with dissociated Fe2O3 and Al2O3 were much higher than those of total Fe and total Al contents, respectively. It was suggested that, in ZFAs, zeolite and iron oxide acted as ion exchanger and adsorbent for Cr(III), respectively, while Ca components elevated the pH of the reaction system and consequently promoted ion exchange and adsorption and caused the surface precipitation of chromium hydroxide. PMID- 18400231 TI - A fluorescent magnetic nanoalloy--Lanthanon-doped FePt:RE. AB - Employing dibenzo-24-crown-8-ether (DB24C8) as a phase-transfer catalyst, monodispersed fluorescent lanthanon-doped magnetic FePt:RE (RE=Eu, Dy, and Ce) nanoparticles about 3 nm in size were synthesized through the reduction of H2PtCl6.6H2O, Fe2(C2O4)3.5H2O, and RE(NO3)3 (RE=Eu, Dy, and Ce) by propylene glycol using oleic acid as the stabilizer in the solvent-thermal system. The conversion of the as-synthesized chemically disordered fcc FePt:RE nanoparticles to a chemically ordered L1 0 structure occurred after annealing treatment at 873 K, and was simultaneously accompanied by a coercivity increase. It is interesting that the amorphous formation trend is strengthened in an europium-doped FePt:Eu alloy accompanied by enhancement of the coercive force. Its thermal stability indicated that the addition of europium can inhibit the phase transformation. Moreover, the optical measurement results proved that FePt:Dy alloy nanoparticles have fluorescent properties. PMID- 18400232 TI - Preparation and optical properties of worm-like gold nanorods. AB - A type of worm-like nanorods was successfully synthesized through conventional gold nanorods reacting with Na2S2O3 or Na2S. The generated worm-like gold nanorods comprise shrunk nanorod cores and enwrapped shells. Therefore, a gold gold sulfide core-shell structure is formed in the process, distinguishing from their original counterparts. The formation of the gold chalcogenide layers was confirmed by transmission electron microscopy and X-ray photoelectron spectroscopy. Experimental results showed that the thickness of the gold chalcogenide layers is controllable. Since the increase of shell thickness and decrease of gold nanorod core take place simultaneously, it allows one to tune the plasmon resonance of nanorods. Proper adjustment of reaction time, temperature, additives and other experimental conditions will produce worm-like gold nanorods demonstrating desired longitudinal plasmon wavelength (LPW) with narrow size distributions, only limited by properties of starting original gold nanorods. The approach presented herein is capable of selectively changing LPW of the gold nanorods. Additionally, the formed worm-like nanorods possess higher sensitive property in localized surface plasmon resonance than the original nanorods. Their special properties were characterized by spectroscopic methods such as Vis-NIR, fluorescence and resonance light scattering. These features imply that the gold nanorods have potential applications in biomolecular recognition study and biosensor fabrications. PMID- 18400233 TI - A sensitive three-step protocol for fluorescence-based Western blot detection. AB - Western blotting is widely used in protein analysis, classically using enhanced chemoluminescence for protein detection. Fluorescence-labelled secondary antibodies have emerged in recent years for detection of antigens, in order to improve the sensitivity and the linear range of detection. Here we show that the sensitivity can be further improved by an additional step in the detection procedure: the antigen is detected by successive incubations with a primary antibody, followed by a biotinylated secondary antibody and then a tertiary fluorescent conjugate. Using the detection of different antigens by CyDye conjugated secondary antibodies in a two-step protocol as a reference, two tertiary fluorescent conjugates were evaluated: CyDye-conjugated streptavidin and CyDye-conjugated anti-biotin antibody. An four-fold increase in sensitivity was achieved with CyDye-conjugated streptavidin; numerous unspecific bands were also generated. CyDye-conjugated anti-biotin antibody did not generate any unspecific bands and led to a 30-fold increase in sensitivity, compared to detection with CyDye-conjugated secondary antibody. PMID- 18400234 TI - Structure of the cytoskeleton of Spiroplasma melliferum BC3 and its interactions with the cell membrane. AB - Spiroplasma melliferum is a wall-less bacterium with dynamic helical symmetry. Taking advantage of the simplicity of this primitive lifeform, we have used structural (electron tomography and freeze fracture of whole cells; cryoelectron tomography and diffraction analysis of isolated cytoskeletons) and proteomic approaches to elucidate the basic organizing principles of its minimal yet functional cytoskeleton. From among approximately 30 Spiroplasma proteins present in a highly purified cytoskeletal fraction, we identify three major putative structural proteins: Fib, MreB, and elongation factor Tu. Fib assembles into a single flattened ribbon that follows the shortest helical line just under the plasma membrane and acts as a linear motor, whereas MreB is present as a matching array of membrane-associated fibrils parallel and associated with the motor. We also identify a prominent previously unknown filamentous network that occupies much of the cytoplasm and appears to cross-link the ribosomes. The abundant potentially filament-forming protein elongation factor Tu may be a component of this network, but the tomography data are most consistent with DNA as the core component. The results provide new information on the minimal organization necessary to support the scaffolding and motile functions of a minimal cytoskeleton. PMID- 18400235 TI - Simvastatin prevents cardiac hypertrophy in vitro and in vivo via JAK/STAT pathway. AB - Simvastatin (SIM), a HMG-CoA reductase inhibitor, has therapeutic effects that are not limited to cholesterol reduction. In this study, we investigated the change in the cell surface area and protein content of cultured rat cardiomyocytes on exposure to cardiotrophin-1 (CT-1), a cytokine involved in the growth and survival of cardiac cells, plus SIM, and thus confirmed that SIM ameliorated cardiomyocyte hypertrophy induced by CT-1. We also showed that SIM attenuated cardiac hypertrophy in rats with pressure overload due to abdominal aortic constriction by measuring such parameters as systolic blood pressure, ratio of heart weight to body weight and ratio of left ventricular weight to body weight in rats as well as cross-sectional area of cardiomyocytes. Western blot analysis indicated that the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway was involved in the mechanisms underlying the in vitro and in vivo inhibitory effects of SIM on cardiac hypertrophy. Moreover, the effect of SIM amelioration on CT-1-induced cultured cardiomyocyte hypertrophy might be related to the change in angiotensinogen (AGT) mRNA expression, as evidenced by RT-PCR analysis, and the subsequent alteration in angiotensin II (Ang II) levels. The results of our study provide further evidence that SIM, like other HMG-CoA reductase inhibitors, is a promising drug for prevention and treatment of cardiac hypertrophy. PMID- 18400236 TI - Statistical approach in search for optimal signal in simple olfactory neuronal models. AB - Several models (concentration detectors and a flux detector) for coding of odor intensity in olfactory sensory neurons are investigated. Behavior of the system is described by different stochastic processes of binding the odorant molecules to the receptors and their activation. Characteristics how well the odorant concentration can be estimated from the knowledge of response, the number of activated neurons, are studied. The approach is based on the Fisher information and analogous measures. These measures of optimality are computed and applied to locate the odorant concentration which is most suitable for coding. The results are compared with the classical deterministic approach which judges the optimal odorant concentration via steepness of the input-output function. PMID- 18400237 TI - Bioactive metabolites from the endophytic fungus Ampelomyces sp. isolated from the medicinal plant Urospermum picroides. AB - Extracts of cultures grown in liquid or on solid rice media of the fungal endophyte Ampelomyces sp. isolated from the medicinal plant Urospermum picroides exhibited considerable cytotoxic activity when tested in vitro against L5178Y cells. Chromatographic separation yielded 14 natural products that were unequivocally identified based on their 1H and 13C NMR as well as mass spectra and comparison with previously published data. Six compounds (2, 4, 5, 7, 9 and 11) were natural products. Both fungal extracts differed considerably in their secondary metabolites. The extract obtained from liquid cultures afforded a pyrone (2) and sulfated anthraquinones (7 and 9) along with the known compounds 1, 3, 6 and 8. When grown on solid rice medium the fungus yielded three compounds 4, 5 and 11 in addition to several known metabolites including 6, 8, 10, 12, 13 and 14. Compounds 4, 8 and 10 showed the strongest cytotoxic activity against L5178Y cells with EC50 values ranging from 0.2-7.3microg/ml. Furthermore, 8 and 10 displayed antimicrobial activity against the Gram-positive pathogens, Staphylococcus aureus, S. epidermidis and Enterococcus faecalis at minimal inhibitory concentrations (MIC) of 12.5microg/ml and 12.5-25microg/ml, respectively. Interestingly, 6 and 8 were also identified as constituents of an extract derived from a healthy plant sample of the host plant U. picroides thereby indicating that the production of bioactive natural products by the endophyte proceeds also under in situ conditions within the host plant. PMID- 18400238 TI - Monoterpene glycosides from Paeonia hybrida. AB - Monoterpene glycosides (1-5, and 7), together with 14 known compounds, were isolated from the methanol extract of the roots of Paeonia hybrida. These compounds included a paeoniflorin-related glycoside with a hybrid structure of paeoniflorin and paeonovicinoside (1), a monoterpene glucoside biogenetically related to lactiflorin (2), a paeoniflorin-related monoterpene (3), arbiflorin related monoterpenes (4 and 5), and a tymol-related monoterpene glycoside (7). Their structures were elucidated by extensive spectroscopic examinations. PMID- 18400239 TI - Bioactive aristolactams from Piper umbellatum. AB - Four alkaloids named piperumbellactams A-D (1-4) were isolated from branches of Piper umbellatum together with known N-hydroxyaristolam II (5), N-p-coumaroyl tyramine (6), 4-nerolidylcatechol (7), N-trans-feruloyltyramine, E-3-(3,4 dihydroxyphenyl)-N-2-[4-hydroxyphenylethyl]-2-propenamide, beta-amyrin, friedelin, apigenin 8-C-neohesperidoside, acacetin 6-C-beta-d-glucopyranoside, beta-sitosterol, its 3-O-beta-d-glucopyranoside and its 3-O-beta-d-[6' dodecanoyl]-glucopyranoside. Glycosidase inhibition, antioxidant and antifungal activities of these compounds were evaluated. Compounds 1-3 showed moderate alpha glucosidase enzyme inhibition with IC50 values 98.07+/-0.44, 43.80+/-0.56 and 29.64+/-0.46, respectively. In DPPH radical scavenging assay, compounds 2, 3 and 6 showed potent inhibitory activity while compounds 4, 5 and 7 showed potent antifungal activity. PMID- 18400240 TI - Outbreak of Chikungunya fever in Mayotte, Comoros archipelago, 2005-2006. AB - In 2005-2006, a large outbreak of Chikungunya (CHIK) fever occurred on the western Indian Ocean Islands. In Mayotte, concurrent with an enhanced passive case notification system, we carried out two surveys. A seroprevalence survey designed to document recent CHIK infection was conducted on serum samples collected from pregnant women in October 2005 (n=316) and in March-April 2006 (n=629). A cross-sectional clinical community survey carried out from 2 to 10 May 2006 among 2235 individuals was designed to determine the cumulative incidence of presumptive CHIK fever cases. The seroprevalence of recent infection among pregnant women was 1.6% in October 2005 and rose to 26% in April 2006. The clinical community survey showed that nearly 26% of respondents had experienced presumptive CHIK fever between January and May 2006. Extrapolated to the overall population of Mayotte, these figures lead to an estimated attack rate of 249.5 cases per 1000 population as of early May 2006. Nine patients with the maternofetal form and six subjects with the severe form were recorded. This first emergence of CHIK fever in Mayotte lead to a very large outbreak. Efforts to strengthen surveillance and prevention of arbovirus infection are needed at country and regional levels. PMID- 18400241 TI - From topical antidote against skin irritants to a novel counter-irritating and anti-inflammatory peptide. AB - The primary purpose of the present study was to investigate the mechanism of the counter-irritating activity of topical iodine against skin lesions induced by chemical and thermal stimuli. The hypothesis that iodine exerts its activity by inducing an endogenous anti-inflammatory factor was confirmed by exposing guinea pig skin to heat stimulus followed by topical iodine treatment and skin extraction. Injection of the extract into naive guinea pigs reduced heat-induced irritation by 69%. The protective factor, identified as a new nonapeptide (histone H2A 36-44, H-Lys-Gly-Asn-Tyr-Ala-Glu-Arg-Ileu-Ala-OH), caused reduction of 40% in irritation score in heat-exposed guinea pigs. The murine analog (H-Lys Gly-His-Tyr-Ala-Glu-Arg-Val-Gly-OH, termed IIIM1) reduced sulfur mustard (SM) induced ear swelling at a dose-dependent bell-shape manner reaching peak activity of 1 mg/kg. Cultured keratinocytes transfected with the peptide were more resistant towards SM than the control cells. The peptide suppressed oxidative burst in activated neutrophils in a concentration-dependent manner. In addition, the peptide reduced glucose oxidase-induced skin edema in mice at a dose dependent bell-shape manner. Apart from thermal and chemical-induced skin irritation this novel peptide might be of potential use in chronic dermal disorders such as psoriasis and pemphigus as well as non-dermal inflammatory diseases like multiple sclerosis, arthritis and colitis. PMID- 18400242 TI - Salmon trypsin stimulates the expression of interleukin-8 via protease-activated receptor-2. AB - In this study, we focus on salmon trypsin as an activator of inflammatory responses in airway cells in vitro. The rationale behind the investigation is that salmon industry workers are exposed to aerosols containing enzymes, which are generated during industrial processing of the fish. Knowing that serine proteases such as trypsin are highly active mediators with diverse biological activities, the stimulation of nuclear factor-kappa B (NF-kappaB) and interleukin (IL)-8 and the role of protease-activated receptors (PAR) in inflammatory signal mediation were investigated. Protease-activated receptors are considered important under pathological situations in the human airways, and a thorough understanding of PAR-induced cellular events and their consequences in airway inflammation is necessary. Human airway epithelial cells (A549) were exposed to trypsin isolated from fish (Salmo salar), and we observed that purified salmon trypsin could generate secretion of IL-8 in a concentration-dependent manner. Furthermore, we demonstrate that PAR-2 activation by salmon trypsin is coupled to an induction of NF-kappaB-mediated transcription using a PAR-2 transfected HeLa cell model. Finally, we show that the release of IL-8 from A549 following stimulation with purified salmon trypsin is mediated through activation of PAR-2 using specific small interfering RNAs (siRNAs). The results presented suggest that salmon trypsin, via activation of PAR-2, might influence inflammation processes in the airways if inhaled in sufficient amounts. PMID- 18400243 TI - Susceptibility to virus-cell fusion at the plasma membrane is reduced through expression of HIV gp41 cytoplasmic domains. AB - The cytoplasmic tail of the HIV transmembrane protein plays an important role in viral infection. In this study we analyzed the role of retroviral cytoplasmic tails in modulating the cytoskeleton and interfering with virus-cell fusion. HeLaP4 cells expressing different HIV cytoplasmic tail constructs showed reduced acetylated tubulin levels whereas the cytoplasmic tail of MLV did not alter microtubule stability indicating a unique function for the lentiviral cytoplasmic tail. The effect on tubulin is mediated through the membrane proximal region of the HIV cytoplasmic tail and was independent of membrane localization. Site directed mutagenesis identified three motifs in the HIV-2 cytoplasmic tail required to effect the reduction in acetylated tubulin. Both the YxxPhi domain and amino acids 21 to 45 of the HIV-2 cytoplasmic tail need to be present to change the level of acetylated tubulin in transfected cells. T-cells stably expressing one HIV-2 cytoplasmic tail derived construct showed also a reduction in acetylated tubulin thus confirming the importance of this effect not only for HeLaP4 and 293T cells. Challenge experiments using transiently transfected HeLaP4 cells and T cells stably expressing an HIV cytoplasmic tail construct revealed both reduced virus-cell fusion and replication of HIV-1(NL4.3) compared to control cells. In the virus-cell fusion assay only virions pseudotyped with either HIV or MLV envelopes showed reduced fusion efficiency, whereas VSV-G pseudotyped virions where not affected by the expression of HIV derived cytoplasmic tail constructs, indicating that fusion at the plasma but not endosomal membrane is affected. Overexpression of human histone-deacetylase 6 (HDAC6) and constitutively active RhoA resulted in a reduction of acetylated tubulin and reduced virus-cell fusion as significant as that observed following expression of HIV cytoplasmic tail constructs. Inhibition of HDAC6 showed a strong increase in acetylated tubulin and increase of virus-cell fusion confirming the correlation between post-translational modification of tubulin and virus-cell fusion. These results thus identify tubulin and its post-translational modification as a new cellular target for interference with HIV-cell fusion. PMID- 18400244 TI - Reaction of [Pt(Gly-Gly-N,N',O)I]- with the N-acetylated dipeptide L-methionyl-L histidine: selective platination of the histidine side chain by intramolecular migration of the platinum(II) complex. AB - The reaction of the monofunctional [Pt(Gly-Gly-N,N',O)I](-) complex, in which Gly Gly is the dipeptide glycyl-glycine coordinated through two nitrogen and oxygen atoms, with the N-acetylated dipeptide L-methionyl-L-histidine (MeCOMet-His) studied by (1)H NMR spectroscopy. All reactions were carried out in 50mM phosphate buffer at pD 7.4 and at 25 degrees C. In the initial stage of the reaction, the platinum(II) complex forms the kinetically favored [Pt(Gly-Gly N,N',O)(MeCOMet-His-S)](-) complex, with unidentate coordination of the MeCOMet His dipeptide through the sulfur atom of the methionine residue. In the second stage of the reaction, complete intramolecular migration of the [Pt(Gly-Gly N,N',O)] unit from the sulfur to the N3 nitrogen atom of imidazole was observed and a new platinum(II)-peptide complex, [Pt(Gly-Gly-N,N',O)(MeCOMet-His-N3)](-) was formed. In comparison with previous results obtained for the reaction of [Pt(dien)Cl](+) with different methionine- and histidine-containing peptides, this migration reaction was sufficiently fast and strongly selective to the N3 atom of the imidazole ring of the histidine side chain. This study is an important step in the development of new platinum(II) complexes for selective covalent modification of peptides and proteins. PMID- 18400245 TI - Desorption of sediment-associated polychlorinated dibenzo-p-dioxins, dibenzofurans, diphenyl ethers and hydroxydiphenyl ethers from contaminated sediment. AB - The transport and bioavailability of sediment-associated contaminants are often controlled by the contaminants' desorbing behaviour. This study examines the desorption kinetics of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), diphenyl ethers (PCDEs) and hydroxydiphenyl ethers (HO-PCDEs) from the highly contaminated River Kymijoki sediment in Finland. The desorption kinetics data were generated using Tenax((R)) extraction, and a first-order three compartment kinetic model was fitted to the data. The desorption data was compared to the previously published accumulation data from this same location to investigate the relationship between the rapidly desorbing fraction (F(r)) and biota-sediment accumulation factors (BSAFs) as well as semipermeable membrane device sediment accumulation factors (SSAFs). The PCDDs, PCDFs, PCDEs and HO PCDEs were tightly attached to sediment particles and formed a large very slowly desorbing fraction (F(vs)). Rapidly desorbing fractions (F(r)) varied between 0.8% and 8% of total amount in sediment. The size of the desorbing fraction was congener-specific and F(r) decreased with the increasing lipophilicity of congeners. The size of the F(r) was unable to explain the small variation in the BSAFs of Lumbriculus variegatus but may help to explain the observed variation in the SSAFs. To our best knowledge, this study is the first effort to investigate the desorption of PCDDs, PCDFs, PCDEs and HO-PCDEs in field-contaminated sediments. The major finding that the very slow desorption of these chemicals will continue years, provides essential information for the modern risk assessment process. PMID- 18400246 TI - Comments on "Elimination of 10 polybrominated diphenyl ether (PBDE) congeners and selected polychlorinated biphenyls from the freshwater mussel, Elliptio complanata" by Ken G. Drouillard, Stella Chan, Sarah O'Rourke, G. Douglas Haffner, Robert J. Letcher [Chemosphere 69(3) (2007) 362-370]. PMID- 18400247 TI - Aerobic biodegradability of the calcium channel antagonist verapamil and identification of a microbial dead-end transformation product studied by LC MS/MS. AB - In recent years pharmaceuticals and personal care products have been detected in increasing concentrations in hospital effluents, sewage treatment plants (STP) as well as in different environmental compartments such as surface water, groundwater and soil. Little is known about the elimination of these substances during sewage treatment or about the formation of potential metabolites in the environment caused by bacterial biotransformation. To assess the biodegradability of the popular cardiovascular drug verapamil and the possible formation of potential microbial degradation products, two tests from the OECD series were used in the present study: the widely used Closed Bottle test (OECD 301 D) and the modified Zahn-Wellens test (OECD 302 B). In the Closed Bottle test, a screening test that simulates the conditions of an environmental surface water compartment, no biological degradation was observed for verapamil at concentrations of 2.33mgl(-1). In the Zahn-Wellens test, a test for inherent biodegradability which allows evaluation of aerobic degradation at high bacterial density, only a partial biological degradation was found. Analysis of test samples by high performance liquid chromatography coupled to multiple stage mass spectrometry (HPLC-MSn) revealed 2-(3,4-dimethoxyphenyl)-2-isopropyl-5 (methylamino)pentane nitrile, already known as D617 (Knoll nomenclature), a metabolite of mammalian metabolism, which is the major degradation product and dead-end transformation product of aerobic degradation of verapamil. PMID- 18400248 TI - Microtox toxicity test: detoxification of TNT and RDX contaminated solutions by poplar tissue cultures. AB - Poplar (Populus deltoidesxnigra DN34) tissue cultures removed 2,4,6 trinitrotoluene (TNT) from an aqueous solution in five days, reducing the toxicity of the solution from highly toxic Microtox EC value to that of the control. 1,3,5-Trinitro-1,3,5-triazacyclohexane (RDX) was taken up by the plant tissue cultures more slowly, but toxicity reduction of the solution was evident. The measurement of toxicity reduction of aqueous solutions containing TNT and RDX was performed using a novel methodology developed for use with the Microtox testing system. Radiolabeled TNT and RDX were used to confirm removal of explosives from hydroponic solutions containing plant tissue cultures and to verify that toxicity did not change in solutions where no plant cultures were present (positive controls). High Performance Liquid Chromatography (HPLC) and Liquid Scintillation Counter (LSC) measurements confirmed removal of TNT and RDX from solutions containing poplar plant tissue cultures and constancy of the plant free controls. In addition, metabolites were identified in remediated solutions by HPLC, confirming the mechanism by which plants can remediate groundwater, surface water, and soil solutions. PMID- 18400249 TI - Evaluation of municipal sewage treatment systems for pollutant removal efficiency by measuring levels of micropollutants. AB - In order to evaluate the municipal sewage treatment systems used at Harbin municipal sewage treatment plant for their pollutant removal efficiency, raw sewage and effluent samples at different treatment stages from the sewage treatment systems were taken, priority pollutants (PPs) were identified and quantified using gas chromatography-mass spectrometry (GC-MS) and inductively coupled plasma-atomic emission spectrograph (ICP-AES). The test results indicated that there were one hundred and fifty species of organic pollutants identified in the raw sewage sample, and only ten species of PPs in all the sewage samples. The levels of dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), di-n-octyl phthalate (DnOP) in the sewage samples were 0.779-0.111 microg l(-1), 1.977-0.022 microg l(-1), 6.411-2.194 microg l(-1) and 7.152-2.953microg l(-1), respectively, and most of these phthalate esters (PAEs) were removed through anaerobic/aerobic (A/O) process; The levels of alachlor, acetochlor, atrazine were 0.074-0.021 microg l(-1), 0.160-0.096 microg l(-1) and 0.238-0.184 microg l(-1), respectively, and the total removal efficiency of atrazine was poorest through the sewage treatment systems. The levels of Cu, Cr, Se, Hg, Ni and Zn were 0.0030-0.2327 mg l(-1). It is therefore concluded from these results that the sewage treatment systems were efficient in removing most of the organic and inorganic compounds in this study, and so, the discharged effluent could cause little of the secondary pollution of the aquatic environment. PMID- 18400251 TI - Mucinous tubular and spindle cell carcinoma with aggressive histomorphology--a sarcomatoid variant. AB - Mucinous tubular and spindle cell carcinoma (MTSCC) is a recently described renal epithelial tumor. The bland cytomorphology of the spindled component and low grade behavior help in its differentiation from sarcomatoid renal carcinoma. Sarcomatoid change has been reported in most histologic variants of renal cell carcinoma apart from MTSCC. Herein we report a case of an MTSCC in a 72-year-old female patient with high-grade spindled areas resembling fibrosarcomatous and undifferentiated pleomorphic sarcoma patterns with metaplastic bone. This index case also demonstrates a high proliferation index and extensive necrosis representing the first documented case of sarcomatoid change in MTSCC. PMID- 18400250 TI - Association between multidrug resistance-associated protein 1 and poor prognosis in patients with nasopharyngeal carcinoma treated with radiotherapy and concurrent chemotherapy. AB - ATP-binding cassette (ABC) multidrug transporters have been associated with chemoresistance, which is a major obstacle in attempts to improve clinical outcome of patients with nasopharyngeal carcinoma (NPC). In this study, we investigated 3 ABC multidrug transporters including MDR1, MRP1, and BCRP for their potential as prognostic indicators in patients with NPC. We examined the protein expression profiles of MDR1, MRP1, and BCRP in NPC tissues from 60 patients with advanced stages who were treated with radiotherapy and concurrent chemotherapy. The clinicopathologic features, patterns of treatment failure, and survival data were compared with the transporter expression. Univariate analyses were performed to determine the prognostic factors that influenced treatment failure and patient survival. We found that MRP1 expression was strongly predictive of both 5-year survival (P = .025) and disease-free survival (P < .001). However, neither MDR1 nor BCRP expression was correlated with the clinicopathologic parameters. Interestingly, the incidence of recurrence and metastasis for patients in the MRP1-positive group was significantly higher than that in the MRP1-negative group (P = .003). With multivariate analysis, MRP1 expression at the time of diagnosis before the treatment was identified as an independent prognostic factor for both 5-year survival (P = .041) and disease free survival (P = .001). MRP1 expression can therefore be used as a potent molecular risk factor and a guide for chemotherapeutic regimens in patients with advanced stages of NPC. PMID- 18400252 TI - Juvenile juxtacortical chondromyxoid fibroma of bone: a case report. AB - Conventional intramedullary chondromyxoid fibroma (CMF) is a rare benign tumor of bone; juxtacortical lesions are rarer still, and juxtacortical lesions occurring in children are heretofore essentially unreported. We present a case of such a lesion in a 12-year-old boy. This patient, who was previously healthy, presented with a 1-week history of poorly defined pain and mild swelling in the region of the left proximal tibia. Magnetic resonance imaging and bone scan showed changes most consistent with an aggressive biological process. However, the permanent histologic sections showed a (pseudo) lobular pattern of spindle cells with minimal pleomorphism and other features consistent with CMF. A clonal abnormality was detected in 15% of tumor cells karyotyped, characterized by a break in the long arm of chromosome 6 and a balanced Robertsonian translocation involving chromosomes 14 and 21. The patient has remained well and free of recurrence for more than 4 years. In general, CMF needs to be distinguished from its mimicker low-grade chondrosarcoma, and it must be recognized as occurring on bone surfaces among a wide age range of individuals. Juxtacortical CMF has not proven to be unusually aggressive in adults nor in this child, and marginal (en-block) resection remains the treatment of choice. PMID- 18400253 TI - Expression of BRCA1 protein in breast cancer and its prognostic significance. AB - BRCA1 is a tumor suppressor gene which, when mutated, is associated with the development of hereditary breast cancers. In sporadic tumors, although inherent gene mutations are rare, loss of BRCA1, resulting from reduced expression or incorrect subcellular localization, is postulated to be important. The purpose of the current study was to examine the expression and localization of BRCA1 protein and to assess its prognostic value, in a well-characterized series of unselected breast carcinomas. We have examined BRCA1 in a series of invasive breast carcinoma (1940 cases) using tissue microarray and immunohistochemistry, to evaluate its expression pattern and to correlate this with clinicopathologic variables and patient outcome. In breast cancer, complete loss of nuclear expression was observed in 223 cases (15%) and cytoplasmic expression was found in 541 breast cancers (36.6%). Absent or reduced nuclear BRCA1 expression was observed more frequently in ductal carcinoma of no special type and medullary like carcinoma and less frequently in lobular and tubular mixed carcinomas. It was also associated with high-grade, advanced lymph node stage, larger size, vascular invasion, negative estrogen receptor, progesterone receptor and androgen receptor expression, and positive p53 and P-cadherin expression, and with the basal-like class of breast cancer. Altered BRCA1 was associated with shorter disease-free interval. Cytoplasmic expression was also associated with development of recurrence and positive EGFR and HER2 expression. It showed an inverse association with survival particularly in low-grade, small-size, and estrogen receptor-positive subgroups. In the grade 1 subgroup, multivariate analysis with adjustment for other prognostic factors showed that cytoplasmic expression of BRCA1 was an independent predictor of disease-free interval. BRCA1 alteration may play a significant role in the development and progression of breast cancer. Immunohistochemical assessment of BRCA1 expression could provide additional clinically relevant information in routine classification of breast cancer. PMID- 18400254 TI - Inflammatory myofibroblastic tumor and low-grade myofibroblastic sarcoma: a comparative study of clinicopathologic features and further observations on the immunohistochemical profile of myofibroblasts. AB - Inflammatory myofibroblastic tumor (IMT) and low-grade myofibroblastic sarcoma (LGMS) are intermediate- or low-grade malignant myofibroblastic neoplasms. In this study, the clinicopathologic profiles of 24 IMTs and 10 LGMS were compared with a focus on the immunohistochemical profiles of the neoplastic myofibroblasts. The primary antibodies used in this study were specific for the ultrastructural subcellular components: (1) *-smooth muscle actin (*-SMA), muscle specific actin (MSA), calponin, and h-caldesmon for myofilaments; (2) fibronectin for fibronexus; (3) laminin for basal lamina; (4) desmin and cytokeratin for intermediate filaments. Anaplastic lymphoma kinase (ALK) alterations were examined by immunohistochemical means, with selective fluorescence in situ hybridization analysis. Histologically, IMT had inhomogeneous microscopic features with multi-component and multi-patterned architecture, whereas LGMS tended to be more uniform in appearance with a higher cellularity, more prominent nuclear hyperchromasia, and a more widely infiltrative growth pattern than IMT. Immunohistochemically, firstly, more than 90% of the cases of both IMT and LGMS expressed calponin, *-SMA, MSA, and fibronectin, almost all with a high expression level, and no cases were positive for h-caldesmon. Secondly, 33.3% (7/21) of IMTs and 40% (4/10) of LGMS were positive for desmin with a low expression level. The positive percentage for laminin was 81.8% (18/22) in IMT, but was 42.9% (3/7) in LGMS with a low expression level. Thirdly, 13.6% (3/22) of IMTs were positive for cytokeratin, but no expression was found in LGMS. ALK staining was found in 40.9% (9/22) of IMTs, and the presence of ALK gene rearrangements was confirmed by fluorescence in situ hybridization in 5 of 6 IMTs examined. However, neither ALK gene rearrangements nor ALK protein labeling was detected in LGMS (0/9). In summary, IMT and LGMS are both composed of cells displaying well-developed myofibroblastic differentiation, which frequently and extensively express actin-associated proteins (*-SMA, MSA, and calponin) and fibronectin, consistent with the ultrastructure markers (myofilaments and fibronectin fibrils). Laminin expression does not exclude a diagnosis of myofibroblastic neoplasms. ALK and cytokeratin, when positive, can be helpful in differentiating IMT from LGMS. LGMS is not a member of the family of ALK-positive tumors. PMID- 18400255 TI - Keratin immunohistochemistry does not contribute to correct lymph node staging in patients with invasive lobular carcinoma. AB - Studies suggest that immunohistochemistry improves rate of detecting sentinel lymph node metastases and is needed for adequate staging in invasive lobular carcinoma. Our study evaluates the use of cytokeratin immunohistochemistry in detecting sentinel lymph node metastases and its effect on staging patients with invasive lobular carcinoma. Material from 76 patients with invasive lobular carcinoma was reviewed. Cytokeratin immunostaining was performed on negative nodes, and deposits were classified as macrometastasis (>2.0 mm), micrometastasis (>0.2-2 mm), or isolated tumor cells (95%) coverage with the MMR I vaccine in the future. If the vaccination coverage decreases to levels around 90% the anticipation of the first dose of the MMR I from 15 to 12 months of age, will ensure that R(e) remains below 1. PMID- 18400342 TI - Immunization strategies against visceral leishmaniosis with the nucleosomal histones of Leishmania infantum encoded in DNA vaccine or pulsed in dendritic cells. AB - Immunization of BALB/c mice with a DNA vaccine encoding the nucleosomal histones from Leishmania infantum resulted in a complete failure of protection against visceral leishmaniosis (VL), whereas the adoptive transfer of bone marrow-derived dendritic cells pulsed with the same pathoantigens plays an essential role in controlling parasite growth in half of the cases. Reduction of the visceral parasite burden seems to be related to low persistence of regulatory T-cells in the spleen from vaccinated mice. These results provide clues for the optimization of this vaccine strategy with the four Leishmania nucleosomal histones against L. infantum infection. PMID- 18400343 TI - The effect of pre-existing immunity on the capacity of influenza virosomes to induce cytotoxic T lymphocyte activity. AB - Protein antigens encapsulated in virosomes generated from influenza virus can induce antigen-specific cytotoxic T lymphocyte (CTL) responses. In the present study we determined, in a murine model system, whether pre-existing immunity against influenza virus hampers the induction of a CTL response. CTL induction was only slightly reduced by pre-injection of influenza virus-specific antibodies or pre-exposure to influenza virus. Both pretreatments resulted in the same level of reduction, suggesting that virus-specific antibodies rather than T cell responses account for the reduction. Furthermore, a booster immunization enhanced CTL activation, indicating that virosome-specific immunity induced by a prime immunization does not hamper the booster effect. In conclusion, CTL induction against virosome-encapsulated protein antigens is not significantly inhibited by pre-existing humoral or cellular immunity against influenza virus. PMID- 18400344 TI - Assessing the impact of different BCG vaccination strategies on severe childhood TB in low-intermediate prevalence settings. AB - The decrease in overall incidence of TB in industrialised countries, together with the increasing concern for adverse events following BCG immunization, has led to important modifications of BCG policies in the last decades. This article adapts and validates--with surveillance data--a theoretical model estimating the impact of different national BCG vaccination policies on severe childhood TB in low to intermediate TB prevalent countries. The model shows that a universal BCG programme could be beneficial in settings with prevalence levels around 30 sputum smear positive per 100,000. In settings with prevalence levels below 15 per 100,000 the benefit of universal BCG vaccination should be carefully assessed, particularly where prevalence is below 5 per 100,000 and universal vaccination might lead to an excess of adverse events per case prevented. To this purpose the model also provides a tool to assess the theoretical impact of a policy change towards selective 'high-risk groups' vaccination. PMID- 18400346 TI - Oral mucosal response to exposure to diquat: a rare occupational injury. AB - Diquat is a quaternary ammonium herbicide closely related to paraquat, and is used commonly by commercial and domestic gardeners. The systemic toxicity of this group of compounds is well-known, but isolated human oral mucosal responses to topical exposure are not well-reported. We present details of an accidental mucosal exposure to diquat during manufacture and the resultant injury, and give guidance for appropriate management of such an exposure. PMID- 18400345 TI - Prime-boost vaccination with a combination of proteosome-degradable and wild-type forms of two influenza proteins leads to augmented CTL response. AB - Targeting viral antigens for proteosomal degradation has previously been proposed as a means for immunogenicity augmentation. However, utilization of modified unstable antigens may be insufficient for potent T-cell cross-presentation by APCs, a mechanism that requires high levels of the antigenic protein. Therefore, we hypothesized that a recombinant vaccine utilizing a combination of proteosome sensitive and proteosome-resistant versions of an antigen in a prime-boost regimen may provide the most efficient CTL response. To address this hypothesis, we utilized conserved proteosome-resistant influenza A virus proteins M1 and NS1. Unstable versions of these polypeptides were constructed by destroying their 3D structure via truncations or short insertions into predicted alpha-helical structures. These modified polypeptides were stabilized in the presence of the proteosome inhibitor MG132, strongly suggesting that they are degraded via a ubiquitin-proteosome pathway. Importantly, with both M1 and NS1antigens, homologous DNA vaccination with a mixture of unstable and proteosome-resistant wt forms of these proteins resulted in significantly higher CTL activity than vaccination with either wt or degradable forms. The most dramatic effect was seen with NS1, where homologous immunization with a mixture of these two forms was the only regimen that produced a notable elevation of CTL response, compared to vaccination with the wt NS1. Additionally, for M1 protein, heterologous vaccination utilizing the unstable form as prime and wild-type form as boost, demonstrated significant augmentation of the CTL response. These data indicate that combining proteosome-sensitive and proteosome-resistant forms of an antigen during vaccination is advantageous. PMID- 18400347 TI - Individuals with mechanical ankle instability exhibit different motion patterns than those with functional ankle instability and ankle sprain copers. AB - BACKGROUND: Chronic ankle instability commonly develops after lateral ankle sprain, but differences in movement patterns between subtypes of ankle instability have not been determined. We hypothesized mechanically and functionally unstable ankle subjects would demonstrate different kinematics and kinetics compared to a group of ankle sprain injury "copers". METHODS: Sixty three recreational athletes, 21 in each of 3 groups (11 males, 10 females) matched for gender, age, height, mass, and limb dominance were tested. Knee and ankle sagittal and frontal plane motion, peak ground reaction forces, and time to peak ground reaction forces were measured during 5 tasks (walk, step down, run, drop jump, and stop jump). FINDINGS: Using 5 one-way Analyses of Variance with Tukey post hoc testing at alpha=0.05, the mechanically unstable group displayed at least one difference in sagittal and/or frontal plane ankle motion at initial contact, maximum, and/or in displacement than the functionally unstable and coper groups in each task except the run (P<0.05). The greatest number of differences occurred in the drop jump and stop jump tasks. The mechanically unstable group exhibited greater frontal plane displacement and maximum eversion, and decreased sagittal plane displacement and maximum plantar flexion in several tasks. INTERPRETATION: The mechanically unstable group demonstrated altered ankle motion patterns when compared to functionally unstable and coper groups. The landing pattern may help avoid ankle sprains, but may also increase risk of ankle joint degeneration. Unstable ankle groups exhibit different movement patterns and may require different treatment regimens. PMID- 18400348 TI - Mercury, cadmium and lead concentrations in different ecophysiological groups of earthworms in forest soils. AB - Bioaccumulation of Hg, Cd and Pb by eight ecophysiologically distinct earthworm species was studied in 27 polluted and uncontaminated forest soils. Lowest tissue concentrations of Hg and Cd occurred in epigeic Lumbricus rubellus and highest in endogeic Octolasion cyaneum. Soils dominated by Dendrodrilus rubidus possess a high potential of risk of Pb biomagnification for secondary predators. Bioconcentration factors (soil-earthworm) followed the sequence ranked Cd>Hg>Pb. Ordination plots of redundancy analysis were used to compare HM concentrations in earthworm tissues with soil, leaf litter and root concentrations and with soil pH and CEC. Different ecological categories of earthworms are exposed to Hg, Cd and Pb in the topsoil by atmospheric deposition and accumulate them in their bodies. Species differences in HM concentrations largely reflect differences in food selectivity and niche separation. PMID- 18400349 TI - Preferences and choices for care and health insurance. AB - Legislation that came into effect in 2006 has dramatically altered the health insurance system in the Netherlands, placing greater emphasis on consumer choice and competition among insurers. The potential for such competition depends largely on consumer preferences for price and quality of service by insurers and quality of affiliated providers. This study provides initial evidence on the preferences of Dutch consumers and how they view trade-offs between various aspects of health insurance product design. A key feature of the analysis is that we compare the responses of high and low risk individuals, where risk is defined by the presence of a costly chronic condition. This contrast is critically important for understanding incentives facing insurers and for identifying potential unanticipated consequences of market competition. The results from our conjoint analysis suggest that not only high risk but also low risk individuals are willing to pay substantially more for insurance products that can be shown to provide better health outcomes. This suggests that insurance products that are more expensive and provide better quality of care may also attract low risk individuals. Therefore, development and dissemination of good, reliable and understandable health plan performance indicators may effectively reduce the problem of adverse selection. PMID- 18400350 TI - The effects of household joblessness on mental health. AB - It is widely assumed that the economic and social costs that unemployment gives rise to must be exacerbated where joblessness is concentrated within families. This hypothesis is tested in this paper. Specifically, data from the first five waves of the Household, Income and Labour Dynamics in Australia Survey (HILDA), a nationally representative household panel survey administered in Australia, are used to test whether jobless individuals score worse on a measure of mental health when they live in households with other jobless people. Consistent with previous research, unemployment is found to be associated with lower levels of mental health. No evidence, however, can be found for any additional disadvantage to the unemployed stemming from living in a jobless household. PMID- 18400351 TI - A neurobehavioral investigation into judgmental processes: effect of bilateral motor behaviors. AB - Two experiments were conducted in order to examine how different bilateral motor activations of the approach and avoidance motivational systems influenced participants' evaluations of valenced stimuli (figurative expressions and pictures of everyday situations). The first Study (Study 1) showed that participants judged valenced expressions according to the motor congruence model put forward by Cretenet and Dru (2004). This may depend on the compatibility of the valenced stimuli with the congruency of the bilateral motor behaviors that involved two unilateral motor behaviors that are congruent to each other. These results were duplicated in Study 2 with the use of valenced pictures taken from the International Affective Picture System. The overall results shed new light on the influence of motor behaviors on judgments, by determining the motor system as operative in evaluative mechanisms, and not merely a simple executive function of higher cognitive processes. PMID- 18400352 TI - One year follow-up of postpartum-onset obsessive-compulsive disorder: a case series. PMID- 18400353 TI - Evaluation of toxicity of green tea catechins with 90-day dietary administration to F344 rats. AB - Green tea catechins (GTC), polyphenols extracted from the stalks and leaves of Camellia sinensis, are found in the different types of tea beverages and as antioxidant additives to many foods, snacks, fats and fatty oils. As a part of their safety assessment, subchronic toxicity was investigated in male and female F344 rats with dietary administration at concentrations of 0 (control), 0.3%, 1.25% and 5.0% for 90 days. The average daily intakes of GTC in each group were 180, 764 and 3525mg/kg body weight/day, respectively for males, and 189, 820 and 3542mg/kg body weight/day, respectively for females. No mortality or obvious clinical signs were observed throughout the experimental period but body weights were reduced from week 1 to the end of the experiment in 5.0% males. In serum biochemistry, alanine transaminase and alkaline phosphatase in 5.0% males and females and aspartate transaminase in 5.0% females were increased, together with the relative liver weights in both sexes receiving 5.0%. Although decreases were evident for total cholesterol in 0.3-5.0% males and triglycerides in 1.25% and 5.0% males and 5.0% females, these changes were not considered to be adverse. Hematology and histopathological observation revealed no GTC-related toxicological changes. Based on above findings, the no observed adverse effect level (NOAEL) of GTC was estimated to be 1.25% (764mg/kg body weight/day for males and 820mg/kg body weight/day for females). PMID- 18400354 TI - Antioxidant activity and bioactive compounds of ten Portuguese regional and commercial almond cultivars. AB - The antioxidant properties of different almond cultivars (cv.), either regional (Casanova, Duro Italiano, Molar, Orelha de Mula and Pegarinhos cv.) or commercial (Ferraduel, Ferranhes, Ferrastar and Guara cv.) were evaluated through several chemical and biochemical assays: DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, reducing power, inhibition of beta-carotene bleaching, inhibition of oxidative hemolysis in erythrocytes, induced by 2,2'-azobis(2 amidinopropane)dihydrochloride (AAPH), and inhibition of thiobarbituric acid reactive substances (TBARS) formation in brain cells, all used as models for the lipid peroxidation damage in biomembranes. The EC50 values were calculated for all the methods in order to evaluate the antioxidant efficiency of each almond cultivar. Bioactive compounds such as phenols and flavonoids were also obtained and correlated to antioxidant activity. The results obtained were quite heterogeneous, revealing significant differences among the cultivars assayed. Duro Italiano cv. revealed better antioxidant properties, presenting lower EC50 values in all assays, and the highest antioxidants contents. The protective effect of this cultivar on erythrocyte biomembrane hemolysis was maintained during 4h. PMID- 18400355 TI - Ghrelin prevents doxorubicin-induced cardiotoxicity through TNF-alpha/NF-kappaB pathways and mitochondrial protective mechanisms. AB - We had reported that increased levels of endogenous ghrelin during the progression of doxorubicin-induced cardiomyopathy and heart failure might provide a compensatory self-protective effect. We investigated which pathway(s) produced these protective effects in vitro. Primary cultured cardiomyocytes were induced with doxorubicin in the presence or absence of ghrelin or a tumor necrosis factor alpha (TNF-alpha) antagonist (etanercept). Ghrelin up-regulated TNF-alpha in a time- and dose-dependent manner. It significantly reduced cell apoptosis and markers of oxidative stress, such as malondialdehyde (MDA) content and lactate dehydrogenase (LDH) activity; it also increased anti-oxidative enzyme activity such as superoxide dismutase (MnSOD) and catalase (CAT), retained mitochondrial membrane potential and energy metabolism compared with doxorubicin alone. Moreover, ghrelin increased mitochondrial anti-apoptosis related gene protein expression such as bcl-2 and MnSOD, reduced cytoplasmic cytochrome C (Cyt C) release and strengthened the activation of NF-kappaB. All these effects were abrogated by etanercept. This suggests ghrelin affects the TNF-alpha/NF-kappaB activation pathways, up-regulating TNF-alpha, to produce anti-oxidative and anti apoptotic effects that protected cardiomyocytes from doxorubicin-induced cytotoxicity. PMID- 18400356 TI - Agentic extraversion as a predictor of effort-related cardiovascular response. AB - The present study examined an extraversion-based extension of the integrative model of cardiovascular effort regulation by Wright and Kirby [Wright, R.A., Kirby, L.D., 2001. Effort determination of cardiovascular response: an integrative analysis with applications in social psychology. In: Zanna, M.P. (Ed.), Advances in Experimental Social Psychology, Academic Press, San Diego, CA, pp. 255-307.]. This model explains cardiovascular effort reactivity in terms of task difficulty, ability appraisal, and success importance. Aggregate measures of cardiovascular variables (alpha-adrenergic, beta-adrenergic, and cholinergic activation components) were used to measure extraversion-based differences in effort. Subjects performed a sequential letter task (n-back verbal working memory task) with four levels of difficulty. Agentic extraverts (n=10) appraised their ability and happiness as significantly higher than introverts (n=10). Introverts showed the expected shark-fin shaped pattern of effort-related cardiovascular reactivity for the alpha-adrenergic and cholinergic activation components. Effort decreased after the moderately difficult 2-back task. Results provide first evidence for an extraversion-based extension of the model and are discussed with regard to mood and resource allocation as possible mechanisms. PMID- 18400357 TI - Effect of raloxifene on the vaginal epithelium of postmenopausal women. AB - OBJECTIVE: The objective was to analyze the effect of raloxifene on the vaginal epithelium of postmenopausal women. STUDY DESIGN: In this non-randomized clinical trial, 80 women (mean age = 60.6 years) were prospectively studied. Forty patients received 60 mg/day of raloxifene (RG), and 40 women constituted a non treated control group (CG), paired by age and time since menopause. The treated group consisted of patients with osteoporosis of the lumbar spine. Those with a diagnosis of infection in the lower genital tract and using hormone therapy (HT) up to 6 months prior to the study were excluded. Vaginal smears were collected at baseline and after 6 months of intervention. The vaginal maturation value (VMV) was determined, and counts of superficial, intermediate and parabasal cells were performed. Smears were analyzed by only one cytopathologist who was blinded to patient data. The t-test, Wilcoxon test, and Chi-Squared test were used in the statistical analysis. RESULTS: The study groups were homogeneous regarding age, time since menopause, parity, HT use, smoking, and body mass index. No statistically significant differences were observed in VMV median values (RG, 39.7 and 35.7; CG, 50.0 and 50.0, respectively) or in the percentage of superficial, intermediate and parabasal cells between the groups at baseline and after 6 months (p>0.05). There was no significant correlation between VMV and age, time since menopause, previous HT use, or body mass index, in either of the groups. CONCLUSION: Treatment with raloxifene for 6 months has no effect on the maturation of the vaginal epithelium in postmenopausal women with osteoporosis. PMID- 18400358 TI - Clinicopathological characteristics of cervical cancer between 2003 and 2005, after the introduction of a national cancer screening program in Slovenia. AB - OBJECTIVE: An organized cervical cancer (CC) screening program was introduced in Slovenia in 2003. With the purpose of clinical audit we analyzed the clinicopathological characteristics of CC patients for the period between 2003 and 2005. STUDY DESIGN: The retrospectively collected data of 450 CC patients were presented at three Advisory Boards of Gynecologic Oncology in Slovenia. They were stratified for stage, tumor characteristics and treatment methods according to gynecologic examination attendance in the last 5 years preceding the diagnosis of CC (attenders vs. nonattenders). RESULTS: In the period of observation, 242 women (53.8%) visited their gynecologists in the 5-year period prior to diagnosis of CC. Squamous cell carcinoma was present in 378 women (84.0%), adenocarcinoma in 45 (10.0%), adenosquamous carcinoma in 24 (5.3%) and other types in 3 women (0.7%). Attenders were significantly more frequently diagnosed with squamous cell carcinoma than nonattenders (chi-square=5.13; P<0.05). Attenders were significantly more frequently diagnosed in stage IA than in stage IB (chi square=22.35; P<0.01). Similarly, in attenders stage I was significantly more frequent than stage II (chi-square=18.81; P<0.01). Pathologic smears of women with CC in the last 5-year period were most frequently evaluated as Pap II in the years 2003 and 2004 (in 39.1 and 26.4% of women, respectively) and as Pap III (in 27.9% of women) in 2005. Surgery was performed in 282 women (62.7%), radiotherapy in 158 (35.1%), symptomatic therapy in 9 (2.0%) and chemotherapy alone in 1 (0.2%) woman. In attenders, surgery alone was the most frequent treatment method (chi-square=91.18; P<0.01). CONCLUSION: Only in attenders a significant redistribution of CC stages in favor of early stages is observed, and in these women more conservative and less extensive treatment methods could be applied. PMID- 18400359 TI - Myeloprotection following cytotoxic damage: the sooner the better. PMID- 18400360 TI - A cascade of Ca(2+)/calmodulin-dependent protein kinases regulates the differentiation and functional activation of murine neutrophils. AB - OBJECTIVE: The function of neutrophils as primary mediators of innate immunity depends on the activity of granule proteins and critical components of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. Expression of their cognate genes is regulated during neutrophil differentiation by a complex network of intracellular signaling pathways. In this study, we have investigated the role of two members of the calcium/calmodulin-dependent protein kinase (CaMK) signaling cascade, CaMK I-like kinase (CKLiK) and CaMKKalpha, in regulating neutrophil differentiation and functional activation. MATERIALS AND METHODS: Mouse myeloid cell lines were used to examine the expression of a CaMK cascade in developing neutrophils and to examine the effects of constitutive activation vs inhibition of CaMKs on neutrophil maturation. RESULTS: Expression of CaMKKalpha was shown to increase during neutrophil differentiation in multiple cell lines, whereas expression of CKLiK increased as multipotent progenitors committed to promyelocytes, but then decreased as cells differentiated into mature neutrophils. Expression of constitutively active CKLiKs did not affect morphologic maturation, but caused dramatic decreases in both respiratory burst responses and chemotaxis. This loss of neutrophil function was accompanied by reduced secondary granule and gp91(phox) gene expression. The CaMK inhibitor KN 93 attenuated cytokine-stimulated proliferative responses in promyelocytic cell lines, and inhibited the respiratory burst. Similar data were observed with the CaMKKalpha inhibitor, STO-609. CONCLUSIONS: Overactivation of a cascade of CaMKs inhibits neutrophil maturation, suggesting that these kinases play an antagonistic role during neutrophil differentiation, but at least one CaMK is required for myeloid cell expansion and functional activation. PMID- 18400361 TI - Socs3 maintains the specificity of biological responses to cytokine signals during granulocyte and macrophage differentiation. AB - Granulocyte colony-stimulating factor (G-CSF) and interleukin-6 (IL-6) play key roles in regulating emergency granulopoiesis and inflammation, and are both negatively regulated by the inducible intracellular protein suppressor of cytokine signaling-3 (Socs3). Mice with Socs3 deleted specifically in hematopoietic cells succumb to severe neutrophil and macrophage-driven inflammation by 1 year of age, and responses to G-CSF are grossly exacerbated. In order to determine which elements of cellular responses to cytokines require Socs3, we have examined the differentiative and proliferative capacity of hematopoietic progenitor cells stimulated by G-CSF and IL-6. The differentiation of Socs3-deficient progenitor cells is skewed toward macrophage production in response to G-CSF or IL-6, whereas wild-type progenitor cells produce mainly neutrophils. The proliferative capacity of Socs3-deficient progenitor cells is greatly enhanced in response to G-CSF at all concentrations, but only at low concentrations for IL-6. Strikingly, synergistic responses to costimulation with stem cell factor and IL-6 (but not G-CSF) are lost at higher concentrations in Socs3-deficient progenitor cells. Cytokine-induced expression of transcriptional regulators including cebpb, Ets2, Bcl3, c-Myc, Jun, and Fosl2 are differentially regulated in Socs3-deficient cells. The tight regulation by Socs3 of signal transducer and activator of transcription 3 phosphorylation and gene transcription after cytokine receptor ligation significantly influences the fate of myeloid progenitor cells. PMID- 18400362 TI - Gene expression profiles of Hodgkin's lymphoma cell lines with different sensitivity to cytotoxic drugs. AB - OBJECTIVE: The prognosis of patients with Hodgkin's lymphoma (HL) has been significantly improved as a result of combination treatment including chemotherapy. However, some patients are refractory to chemotherapy. Therefore, identification of new targets might be useful for development of alternative treatment strategies. In addition, identification of markers associated with chemoresistance can be used to identify patients with increased risk of relapse. MATERIALS AND METHODS: By using high-density DNA microarrays, we analyzed the gene-expression profile of HL-cell lines in comparison to a set of normal tissues. Furthermore, we tested the sensitivity of HL cells for cytotoxic drugs (cisplatin, etoposide, melphalan) and compared the gene-expression profile of chemotherapy-resistant and -sensitive cell lines. Differentially expressed genes were validated by polymerase chain reaction and flow cytometry. RESULTS: In addition to genes with high expression in all cell lines, we observed differences between the gene-expression profile of chemotherapy-resistant and -sensitive cells. Genes upregulated in resistant cells include cytokine receptors (IL5RA, IL13RA1), markers expressed on antigen-presenting cells (CD40, CD80), as well as genes with known association to chemoresistance, e.g., myristoylated alanine-rich protein kinase C substrate. In addition, the tumor antigen PRAME (preferentially expressed antigen in melanoma) was expressed in resistant cell lines only. CONCLUSION: Genes with high expression in HL cells might be potential targets for development of future therapeutic interventions. Expression of tumor antigens together with costimulatory molecules in chemotherapy-resistant HL cells might become targets for cytotoxic T-cell responses against HL cells. PMID- 18400364 TI - Early fetal gene delivery utilizes both central and peripheral mechanisms of tolerance induction. AB - OBJECTIVE: We previously reported the induction of stable immune tolerance following direct injection of retroviral vectors into preimmune fetal sheep. In the present studies, we conduct detailed analysis of the thymus of recipients of in utero gene transfer (IUGT) to delineate the mechanism of the observed immune tolerance and assess the impact of recipient age on this process. MATERIALS AND METHODS: Fetal sheep at varying gestational ages received the MSCV-NeoR-RFP retroviral vector. The thymus was then collected from these animals at 27 to 30 days postinjection and analyzed for evidence of transduction of key immunoregulatory thymic cells. RESULTS: Our results reveal that both thymic epithelial cells (TEC), crucial for presentation of self-antigen during T-cell thymic selection, and the cells comprising the Hassall's corpuscles, which can present antigen directly and also instruct dendritic cells to induce the formation of CD4(+)CD25(+) T-regulatory cells in the thymus, were only efficiently transduced if IUGT was performed early in gestation. CONCLUSIONS: Our findings thus demonstrate, for the first time, that early IUGT can potentially take advantage of multiple tolerogenic avenues in the fetus, transducing both TEC, which promote central tolerance, and Hassall's corpuscles, which induce formation of T regulatory cells that could act to maintain peripheral tolerance to the transgene products. PMID- 18400365 TI - Regression modeling of streamflow, baseflow, and runoff using geographic information systems. AB - Regression models for predicting total streamflow (TSF), baseflow (TBF), and storm runoff (TRO) are needed for water resource planning and management. This study used 54 streams with >20 years of streamflow gaging station records during the period October 1971 to September 2001 in Pennsylvania and partitioned TSF into TBF and TRO. TBF was considered a surrogate of groundwater recharge for basins. Regression models for predicting basin-wide TSF, TBF, and TRO were developed under three scenarios that varied in regression variables used for model development. Regression variables representing basin geomorphological, geological, soil, and climatic characteristics were estimated using geographic information systems. All regression models for TSF, TBF, and TRO had R(2) values >0.94 and reasonable prediction errors. The two best TSF models developed under scenarios 1 and 2 had similar absolute prediction errors. The same was true for the two best TBF models. Therefore, any one of the two best TSF and TBF models could be used for respective flow prediction depending on variable availability. The TRO model developed under scenario 1 had smaller absolute prediction errors than that developed under scenario 2. Simplified Area-alone models developed under scenario 3 might be used when variables for using best models are not available, but had lower R(2) values and higher or more variable prediction errors than the best models. PMID- 18400366 TI - Alternative treatment for septic tank sludge: co-digestion with municipal solid waste in bioreactor landfill simulators. AB - Co-disposal of septic tank sludge had a positive effect on the municipal solid waste (MSW) stabilisation process in Bioreactor Landfill simulators. Co-disposal experiments were carried out using the Bioreactor Landfill approach aiming to solve the environmental problems caused by indiscriminate and inadequate disposal of MSW and especially of septic tank sludge. The simulator receiving septic tank sludge exhibited a 200 days shorter lag-phase as compared to the 350 days required by the control simulator to start the exponential biogas production. Additionally, the simulator with septic sludge apparently retained more moisture (>60% w/w), which enhanced the overall conversion of organic matter hence increasing the biogas production (0.60 m3 biogas kg(-1)VS(converted)) and removal efficiency of 60% for VS from the simulator. Alkaline pH values (pH>8.5) did not inhibit the biogas production; moreover it contributed to reduce partially the negative effects of NH(4)(+) (>2 g L(-1)) due to NH(3) volatilisation thus reducing the nitrogen content of the residues. Associated risks and hazards with septage disposal were practically eliminated as total coliform and faecal coliform contents were reduced by 99% and 100%, respectively at the end of the experiment. These results indicate that co-disposal has two direct benefits, including the safe and environmentally sound disposal of septic tank sludge and an improvement of the overall performance of the Bioreactor Landfill by increasing moisture retention and supplying a more acclimatised bacterial population. PMID- 18400368 TI - Long-term rates of undetectable PSA with initial observation and delayed salvage radiotherapy after radical prostatectomy. AB - BACKGROUND: Randomized trials have shown an improvement in progression-free survival rates with adjuvant radiation therapy (ART) after radical prostatectomy for patients with a high risk of cancer recurrence. Less is known about the relative advantages and disadvantages of initial observation with delayed salvage radiation therapy (SRT). OBJECTIVE: To examine the results of SRT in a large single-surgeon radical prostatectomy series. DESIGN, SETTING, AND PARTICIPANTS: From a radical prostatectomy database, we identified 859 men with positive surgical margins (SM+), extracapsular tumor extension (ECE), or seminal vesicle invasion (SVI) who chose to defer ART. Following a period of initial observation, 192 ultimately received SRT for prostate-specific antigen (PSA) progression. MEASUREMENTS: Survival analysis was performed to examine the outcomes of initial observation followed by SRT. RESULTS AND LIMITATIONS: In patients with SM+/ECE and SVI, the 7-yr PSA progression-free survival rates with observation were 62% and 32%, respectively. Among those who had PSA progression, 56% and 26%, respectively, maintained an undetectable PSA for 5 yr after SRT. The long-term rates of undetectable PSA associated with an SRT strategy were 83% and 50% for men with SM+/ECE and SVI, respectively. In the subset of 716 men who did not receive any hormonal therapy, the corresponding long-term rates of undetectable PSA were 91% and 75%, respectively. CONCLUSIONS: Following radical prostatectomy, initial observation followed by delayed SRT at the time of PSA recurrence is an effective strategy for selected patients with SM+/ECE. Some patients with SVI may also benefit from this strategy. However, additional prospective studies are necessary to further examine the survival outcomes following SRT. PMID- 18400369 TI - Editorial comment on: long-term rates of undetectable psa with initial observation and delayed salvage radiotherapy after radical prostatectomy. PMID- 18400370 TI - Urethroplasty for stricture disease--what is success beyond the voiding function? PMID- 18400371 TI - How should small renal masses be treated today? PMID- 18400372 TI - Kaempferol has osteogenic effect in ovariectomized adult Sprague-Dawley rats. AB - Kaempferol (K), a flavonol, is known to have anti-osteoclastogenic effect. We here show that K, from 0.2 to 5.0 microM, increased mineralized nodules in rat primary osteoblasts. K also significantly attenuated adipocyte formation from bone marrow cells (BMCs). A single oral dose of 1 mg/kg body weight of K in Sprague-Dawley (180-200 g) rats resulted in a peak serum level of 2.04+/-0.8 nM in 30 min (Tmax), suggesting its rapid absorption. The Cmax of K in bone marrow was 0.684 nM after 90 min. Rats were ovariectomized (OVx) along with sham operated rats and left for 4 weeks. Daily oral administration of K (5 mg/kg body weight) was then started to one group of OVx rats, and continued for 10 weeks. K levels were found to be 0.311 and 0.838 nM at the end of 4 and 10 weeks, respectively. K exhibited no estrogenicity at the uterine level. The K-treated group exhibited significantly higher bone mineral density (BMD) in the trabecular regions (femur neck, proximal tibia and vertebrae) and lower serum ALP (bone turnover marker) compared with the OVx rats. The compressive energy of the vertebrae was significantly higher in the OVx+K-treated group compared with the OVx group. K treatment of OVx rats resulted in the increase in osteoprogenitor cells as well as inhibition of adipocyte differentiation from BMCs compared with the OVx group. Together we show that K is non-estrogenic in vivo and exerts bone anabolic activity with attendant inhibition of bone marrow adipogenesis. PMID- 18400373 TI - The role of parathyroid hormone and insulin-like growth factors in hematopoietic niches: physiology and pharmacology. AB - Hematopoietic stem cells (HSC) capable of both self-renewal and differentiation into all blood lineages reside within the bone marrow in specialized microenvironmental niches. While the precise location and composition of these niches largely remains unknown, it is now believed that osteoblasts at the endosteal surface play critical roles. Among the molecules demonstrated to influence the function of these niches are parathyroid hormone (PTH) and the insulin-like growth factors (IGF). Administration of PTH to both mice and men expands the number of bone marrow HSC, and an increase in the number of those cells in peripheral blood following treatment with mobilizing agents. Several molecules downstream of PTH are capable of signaling to HSC, including IGF that appear to regulate both the survival and expansion of hematopoietic stem and progenitor cells. As our current understanding of the role for PTH and IGF in hematopoietic niches is limited, we believe it is important that both their physiological importance and pharmacological potential be more fully investigated. PMID- 18400374 TI - Thyroid hormone regulation of stem cell development during intestinal remodeling. AB - During amphibian metamorphosis the small intestine is remodeled from larval to adult form, analogous to the mammalian intestine. The larval epithelium mostly undergoes apoptosis, while a small number of stem cells appear, actively proliferate, and differentiate into the adult epithelium possessing a cell renewal system. Because amphibian intestinal remodeling is completely controlled by thyroid hormone (T3) through T3 receptors (TRs), it serves as an excellent model for studying the molecular mechanism of the mammalian intestinal development. TRs bind T3 response elements in target genes and have dual functions by interacting with coactivators or corepressors in a T3-dependent manner. A number of T3 response genes have been isolated from the Xenopus laevis intestine. They include signaling molecules, matrix metalloproteinases, and transcription factors. Functional studies have been carried out on many such genes in vitro and in vivo by using transgenic and culture technologies. Here we will review recent findings from such studies with a special emphasis on the adult intestinal stem cells, and discuss the evolutionarily conserved roles of T3 in the epithelial cell-renewal in the vertebrate intestine. PMID- 18400375 TI - Priming with EGFR tyrosine kinase inhibitor and EGF sensitizes ovarian cancer cells to respond to chemotherapeutical drugs. AB - Over-expression of EGFR, as in most cases of ovarian cancer, is associated with advanced-stage disease and poor prognosis. Activation of EGFR signaling pathway is involved in increased cell proliferation, angiogenesis, metastasis and decreased apoptosis. Tyrosine kinase activity is essential for signal transduction and receptor down-regulation. However, we found in this study that tyrosine kinase activity is not necessary in ligand-induced EGFR down-regulation in ovarian cancer cell line CaOV3 cells. EGFR tyrosine kinase inhibitors, such as PD153035, AG1478, as well as non-specific tyrosine kinase inhibitor PP2 cannot reverse EGF-induced down-regulation of EGFR. These findings thus permit us to develop the following exciting but unconventional strategy to sensitize cancer cells, namely, by priming ovarian cancer cells with EGF and EGFR inhibitor PD153035, before chemotherapy. This priming procedure down-regulates EGFR without induction of mitogenic signals such as ERK and PI3K/AKT. EGF plus EGFR inhibitor primed ovarian cancer cells display increased sensitivity to taxol-induced cell death, resistant to EGF-induced cell migration and cell proliferation as well as ERK and PI3K/AKT activation. Further studies showed that PD153035, which does not reverse ligand-induced EGFR down-regulation, blocks EGF-induced EGFR activation as well as EGFR's binding to c-cbl and Grb2. Taken together, we contend that priming with EGFR inhibitors plus EGF inhibits cell signaling pathways leading to cell proliferation and survival, while down-regulating EGFR. This priming approach sensitizes ovarian cancer cells and would ultimately result in better chemotherapeutical outcome. PMID- 18400376 TI - The release of fibroblast growth factor-1 from melanoma cells requires copper ions and is mediated by phosphatidylinositol 3-kinase/Akt intracellular signaling pathway. AB - Melanoma is a highly invasive tumor with elevated mortality rates. Progression and aggressiveness appear related to the achievement of an angiogenic phenotype. Melanoma cells express several angiogenic factors, including fibroblast growth factor (FGF)-1 and FGF-2. The autocrine production and release of FGFs and the subsequent activation of FGF receptors, have a central role in melanoma tumor progression. We demonstrated that FGF-1 is secreted from a human melanoma cell line, A375, under conditions of serum deprivation. The release of FGF-1 is inhibited by the copper chelator ammonium tetrathiomolybdate, suggesting a role of copper in the secretory pathway, and is triggered by activation of phosphatidylinositol 3-kinase (PI3K)/Akt intracellular signaling. Interestingly, overexpression or activation of Akt has been correlated with poor prognosis in melanoma patients. Our data indicate a novel role for Akt in supporting the progression of human melanomas and advocate the need for new treatments targeting PI3K/Akt signaling pathway, to control tumor development and progression. PMID- 18400377 TI - Hydrometallurgical route to recover molybdenum, nickel, cobalt and aluminum from spent hydrotreating catalysts in sulphuric acid medium. AB - This work describes a hydrometallurgical route for processing spent commercial catalysts (CoMo and NiMo/Al2O3), for recovering the active phase and support components. They were initially pre-oxidized (500 degrees C, 5h) in order to eliminate coke and other volatile species present. Pre-oxidized catalysts were dissolved in H2SO4 (9molL-1) at approximately 90 degrees C, and the remaining residues separated from the solution. Molybdenum was recovered by solvent extraction using tertiary amines. Alamine 304 presented the best performance at pH around 1.8. After this step, cobalt (or nickel) was separated by adding aqueous ammonium oxalate in the above pH. Before aluminum recovery, by adding NaOH to the acid solution, phosphorus (H2PO4-) was removed by passing the liquid through a strong anion exchange column. Final wastes occur as neutral and colorless sodium sulphate solutions and the insoluble solid in the acid leachant. The hydrometallurgical route presented in this work generates less final aqueous wastes, as it is not necessary to use alkaline medium during the metal recovery steps. The metals were isolated in very high yields (>98wt.%). PMID- 18400378 TI - Removal of mercury(II) from aqueous media using eucalyptus bark: Kinetic and equilibrium studies. AB - In this study, eucalyptus camaldulensis bark, a forest solid waste, is proposed as a novel material for the removal of mercury(II) from aqueous phase. The operating variables studied were sorbent dosage, ionic strength, stirring speed, temperature, solution pH, contact time, and initial metal concentration. Sorption experiments indicated that the sorption capacity was dependent on operating variables and the process was strongly pH-dependent. Kinetic measurements showed that the process was uniform and rapid. In order to investigate the mechanism of sorption, kinetic data were modeled using the pseudo-first-order and pseudo second-order kinetic equations, and intraparticle diffusion model. Among the kinetic models studied, the pseudo-second-order equation was the best applicable model to describe the sorption process. Equilibrium isotherm data were analyzed using the Langmuir and the Freundlich isotherms. The Langmuir model yields a much better fit than the Freundlich model. Isotherms have also been used to obtain the thermodynamic parameters such as free energy, enthalpy, and entropy of sorption. The maximum sorption capacity was 33.11mgg(-1) at 20 degrees C and the negative value of free energy change indicated the spontaneous nature of sorption. These results demonstrate that eucalyptus bark is very effective in the removal of Hg(II) from aqueous solutions. PMID- 18400379 TI - Application of nanofiltration for the rejection of nickel ions from aqueous solutions and estimation of membrane transport parameters. AB - The present work deals with the application of a thin-film composite polyamide nanofiltration membrane for the rejection of nickel ions from aqueous wastewater. The operating variables studied are feed concentration (5-250ppm), applied pressure (4-20atm), feed flowrate (5-15L/min) and pH (2-8). It is observed that the observed rejection of nickel ions increases with increase in feed pressure and decreases with increase in feed concentration at constant feed flowrate. The maximum observed rejection of the metal is found to be 98% and 92% for an initial feed concentration of 5 and 250ppm, respectively. The effect of pH on the rejection of nickel ions and permeate flux are studied, and found that the variation in pH is having more effect on the latter than the former. The experimental data are analyzed using membrane transport models; combined-film theory-solution-diffusion (CFSD), combined-film theory-Spiegler-Kedem (CFSK) and combined-film theory-finely porous (CFFP) models; to estimate membrane transport parameters and mass transfer coefficient, k. Also, enrichment factor, concentration polarization modulus and Peclet number are found from various parameters. From CFFP model the effective membrane thickness and active skin layer thickness are found. PMID- 18400380 TI - Oxolinic acid photo-oxidation using immobilized TiO(2). AB - This work studied the photocatalysed oxidation of the antibiotic oxolinic acid (OA) in an annular reactor operated with immobilized TiO(2) on sintered glass cylinders (SGC). Experiments were carried out in 1l solution of OA (18 mg l(-1)) at pH 9 with oxygen bubbling. Irradiation was performed with black light (36 W). The reaction was monitored by COD, TOC and average oxidation state (AOS) calculations. The antibacterial activity of intermediates was followed using the inhibition halo technique on Escherichia coli cultures. The initial antibiotic concentration decreases in one order of magnitude after 60 min irradiation, and was completely eliminated at 100 min reaction. The TOC was reduced in 54% and the AOS reach values around +3 indicating the formation of low molecular weight carboxylic acids. The oxidation reaction fit well with the Langmuir-Hinshelwood kinetic model indicating the dependence of reaction rate with initial adsorption step. The antibacterial activity of the solution decreases with antibiotic removal, demonstrating that intermediates do not present antibiotic activity. PMID- 18400381 TI - The removal of an anionic red dye from aqueous solutions using chitosan beads-the role of experimental factors on adsorption using a full factorial design. AB - A factorial design was employed to evaluate the quantitative removal of an anionic red dye from aqueous solutions on epichlorohydrin-cross-linked chitosan. The experimental factors and their respective levels studied were the initial dye concentration in solution (25 or 600mgL(-1)), the absence or the presence of the anionic surfactant sodium dodecylbenzenesulfonate (DBS) and the adsorption temperature (25 or 55 degrees C). The adsorption parameters were analyzed statistically using modeling polynomial equations. The results indicated that increasing the dye concentration from 25 to 600mgL(-1) increases the dye adsorption whereas the presence of DBS increases it. The principal effect of temperature did not show a high statistical significance. The factorial results also demonstrate the existence of statistically significant binary interactions of the experimental factors. The adsorption thermodynamic parameters, namely Delta(ads)H, Delta(ads)G and Delta(ads)S, were determined for all the factorial design results. Exothermic and endothermic values were found in relation to the Delta(ads)H. The positive Delta(ads)S values indicate that entropy is a driving force for adsorption. The Delta(ads)G values are significantly affected by an important synergistic effect of the factors and not by the temperature changes alone. PMID- 18400382 TI - Ferric ion mediated photochemical decomposition of perfluorooctanoic acid (PFOA) by 254nm UV light. AB - The great enhancement of ferric ion on the photochemical decomposition of environmentally persistent perfluorooctanoic acid (PFOA) under 254nm UV light was reported. In the presence of 10microM ferric ion, 47.3% of initial PFOA (48microM) was decomposed and the defluorination ratio reached 15.4% within 4h reaction time. While the degradation and defluorination ratio greatly increased to 80.2% and 47.8%, respectively, when ferric ion concentration increased to 80microM, and the corresponding half-life was shortened to 103min. Though the decomposition rate was significantly lowered under nitrogen atmosphere, PFOA was efficiently decomposed too. Other metal ions like Cu(2+) and Zn(2+) also slightly improved the photochemical decomposition of PFOA under irradiation of 254nm UV light. Besides fluoride ion, other intermediates during PFOA decomposition including formic acid and five shorter-chain perfluorinated carboxylic acids (PFCAs) with C7, C6, C5, C4 and C3, respectively, were identified and quantified by IC or LC/MS. The mixture of PFOA and ferric ion had strong absorption around 280nm. It is proposed that PFOA coordinates with ferric ion to form a complex, and its excitation by 254nm UV light leads to the decomposition of PFOA in a stepwise way. PMID- 18400383 TI - Significance of the long-chain organic cation structure in the sorption of the penconazole and metalaxyl fungicides by organo clays. AB - Sorption-desorption of two different hydrophobic fungicides, penconazole and metalaxyl, by a series of clay minerals layered and non-layered (montmorillonite, illite, kaolinite, muscovite, sepiolite and palygorskite) modified with the two chain cationic surfactant, dihexadecyldimetylammonium (DHDDMA) was studied by first time in this work. DHDDMA-clays showed a good capacity to sorb both fungicides from water. Freundlich sorption constants (K(f)) increased 22-268-fold for penconazole and 4-112-fold for metalaxyl in relation to natural clays. High sorption irreversibility was observed for penconazole for all modified clays, while for metalaxyl this occurred only for some of the modified clays. Additionally, a comparative study of the sorption of the fungicides by clay minerals modified with organic cations of different structure was carried out by a statistical approach. Non-layered and layered clay minerals modified with the two-chain alkylammonium organic cation DHDDMA, and with single-chain organic cations octadecyltrimethylammonium (ODTMA) and hexadecylpyridinium (HDPY) were used as sorbents. The study indicated a significant linear regression between the distribution coefficients K(d) of fungicides and the organic carbon (OC) content of the organo clays (r(2)>or=0.80, p<0.001). According to this sorption of fungicides by partition between the aqueous solution and the organic medium created by the alkyl chains of the exchanged ammonium cations regardless of the type and structure of these cations is suggested. However, linear relationships between K(d) values of each pesticide by the different organo clays and their relative OC contents revealed higher increase in sorption for HDPY-clays (penconazole) and for ODTMA-clays (metalaxyl) pointing out different effectiveness of the OC provided by each organic cation. These effects were also supported by the values of K(d) relative to OC, K(d)(oc). The results evidenced the different relative weight of alkyl cations to make organo clays effective barriers to prevent the mobility of pesticides from a point source of pollution. PMID- 18400384 TI - Enhanced activity of bismuth-compounded TiO(2) nanoparticles for photocatalytically degrading rhodamine B solution. AB - TiO(2) nanoparticles compounded with different amounts of bismuth were prepared by a sol-gel method, and the effects of compounding bismuth on the phase transformation, photoinduced charge separation and photocatalytic activity for degrading rhodamine B solution were mainly investigated, along with enhancement mechanism of photocatalytic activity of TiO(2) nanoparticles by compounding bismuth species. It can be confirmed that, by means of X-ray diffraction (XRD), surface photovoltage spectroscopy (SPS) and ultraviolet-visible diffuse reflectance spectroscopy (UV-vis DRS), compounding bismuth can extend the optical response, and effectively inhibit the phase transformation process from anatase to rutile, consequently greatly improving the anatase crystallinity so as to promote the photoinduced charge separation. These factors are responsible for the increase in the photocatalytic activity of TiO(2) compounded with an appropriate amount of bismuth species. PMID- 18400385 TI - Adsorption properties of Congo Red from aqueous solution onto surfactant-modified montmorillonite. AB - A series of surfactant-modified montmorillonites (MMT) were prepared using octyltrimethylammonium bromide (OTAB), dodecyltrimethylammonium bromide (DTAB), cetyltrimethylammonium bromide (CTAB) and stearyltrimethylammonium bromide (STAB), and the organification of MMT was proved by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), scanning electron micrographic (SEM) and transmission electron microscope (TEM). The adsorption of Congo Red (CR) anionic dye from aqueous solution onto surfactant-modified MMT was carried out. Compared with MMT, the adsorption capacity of surfactant-modified MMT for CR was greatly enhanced and MMT modified with CTAB (2.0 CEC) exhibited the higher adsorption capacity. The effects of pH value of the dye solution, adsorption temperature, adsorption time and the initial dye concentration on the adsorption capacity of CR on CTAB-MMT have been investigated. The results showed that the adsorption kinetic of CR on CTAB-MMT could be best described by the pseudo-second order model and that the adsorption isotherm of CR was in good agreement with the Langmuir equation. The IR spectra and SEM analysis also revealed that the adsorption of CTAB-MMT was a chemical adsorption process between CTAB and the NH(2), -N=N- and SO(3) groups of CR. PMID- 18400386 TI - Development of enhanced sulphidogenesis process for the treatment of wastewater having low COD/SO(4)(2-) ratio. AB - An upflow hybrid sulphidogenesis reactor of 1.75 L volume was developed (at oxidation-reduction potential (ORP)=-225+/-25 mV) using flocculent extended aeration process sludge (selected based on screening study at COD/SO(4)(2-) ratio=1) for enhanced sulphidogenesis and COD removal. The reactor was subjected to various loading rate studies at a hydraulic retention time (HRT) of 1 day with COD/SO(4)(2-) ratio of 1.3. At loading rate of 2.5 kg COD/(m(3)day), excellent performance with more than 97% removal of sulphate was achieved within bottom 40% volume of the reactor. At a higher loading rate of 3.75 kg COD/(m(3)day), there was a decrease in both sulphate (70-75%) and COD (50%) removal efficiencies. A controlled and continuous air injection (0.19 L/(L min)) given at 40% volume of the reactor affected sulphide oxidation inside the reactor and enhanced the sulphate reduction in the reactor. The specific sulphate reduction capacity of mixed culture drawn from the bottom part of the reactor was 0.35 kg SO(4)(2-)/(kg VSS day). The results of this study showed that enhanced sulphidogenesis with sulphide inhibition control can maintain sulphate-reducing bacteria (SRB) in anaerobic reactor at low COD/SO(4)(2-) ratios between 1 and 2, with efficient simultaneous removal of COD and SO(4)(2-). The sulphide generated in the system can be recovered as elemental sulphur and/or oxidized back to sulphate. PMID- 18400387 TI - DFT study of the interaction between 3-nitro-1,2,4-triazole-5-one and hydrogen fluoride. AB - Three fully optimized geometries of 3-nitro-1,2,4-triazol-5-one-hydrogen fluoride (NTO-HF) complexes have been obtained with density functional theory (DFT) method at the B3LYP/6-311++G** level. The intermolecular interaction energy is calculated with zero point energy (ZPE) correction and basis set superposition error (BSSE) correction. The greatest corrected intermolecular interaction of the NTO-HF complexes is -34.155kJ/mol. Electrons in complex systems transfer from NTO to HF. Natural bond orbital (NBO) analysis is performed to reveal the origin of the interaction. The strong hydrogen bonds contribute to the interaction energies dominantly. Frequency calculations are carried out on each optimized structure, and their IR spectra are discussed. Vibrational analysis show that there are large red-shifts for H-X (X=N and F) stretching vibrational frequencies in the NTO and hydrogen fluoride complexes. The changes of thermodynamic properties from the monomer to complexes with the temperature ranging from 200K to 1500K have been obtained using the statistical thermodynamic method. It is found that two of three NTO-HF complexes can be produced spontaneously from NTO and HF at room temperature. PMID- 18400388 TI - Predicting activation energy of thermolysis of polynitro arenes through molecular structure. AB - The paper presents a new method for activation energy or the Arrhenius parameter E(a) of the thermolysis in the condensed state for different polynitro arenes as an important class of energetic molecules. The methodology assumes that E(a) of a polynitro arene with general formula C(a)H(b)N(c)O(d) can be expressed as a function of optimized elemental composition as well as the contribution of specific molecular structural parameters. The new method can predict E(a) of the thermolysis under conditions of Soviet Manometric Method (SMM), which can be related to the other convenient methods. The new correlation has the root mean square (rms) and the average deviations of 13.79 and 11.94kJ/mol, respectively, for 20 polynitro arenes with different molecular structures. The proposed new method can also be used to predict E(a) of three polynitro arenes, i.e. 2,2',2'',4,4',4'',6,6',6''-nonanitro-1,1':3',1''-terphenyl (NONA), 3,3'-diamino 2,2',4,4',6,6'-hexanitro-1,1'-biphenyl-3,3'-diamine (DIPAM) and N,N-bis(2,4 dinitrophenyl)-2,4,6-trinitroaniline (NTFA), which have complex molecular structures. PMID- 18400389 TI - Preparation, characterization and photocatalytic activity of manganese doped TiO(2) immobilized on silica gel. AB - A series of Mn-TiO(2)/SiO(2) (silica gel loaded with manganese doped TiO(2)) photocatalysts have been prepared by sol-gel method, and characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy (XPS). Photocatalytic activities were enhanced in photocatalytic degradation of methyl orange over Mn-TiO(2)/SiO(2). XPS analysis shows that a Ti O-Si or Ti-O-Mn bond is formed on the surface of photocatalyst. Mn is doped as a mixture of Mn(2+) and Mn(3+) on the surface of 1.0mol% Mn-TiO(2)/SiO(2). Mn(3+) appears to trap electrons and prohibit the electron-hole recombination. The electrons trapped in Mn(3+) site are subsequently transferred to the adsorbed O(2). As a result, the combination of the electron-hole pair was reduced. PMID- 18400390 TI - Catalytic ozonation of phenol in water with natural brucite and magnesia. AB - Natural brucite and magnesia were applied as catalysts in catalytic ozonation of phenol in this work. It was found that both brucite and magnesia had remarkable accelerations on degradation of phenol and removal of COD in water. On this basis, effective and feasible routes for catalytic ozonation of phenol in water were proposed. The influence of initial pH value, radical scavengers and reaction temperature were investigated. The results revealed that there were different ozonation mechanisms in two systems: molecular ozone direct oxidation mechanism was proved in catalytic ozonation with brucite, and hydroxyl radical mechanism was demonstrated to play a main role in catalytic ozonation with magnesia. PMID- 18400391 TI - Analysis of SUMO-1 modification of neuronal proteins containing consensus SUMOylation motifs. AB - SUMOylation is emerging as an important mechanism for modulating protein function in many cell types. A large variety of proteins have been proposed as SUMO targets based on the presence of a consensus SUMOylation core motif (Psi-K-x D/E). In neurons these include multiple synaptic proteins but it has not been established whether proteins carrying this motif are SUMOylated either in vitro or in vivo. Here we use a bacterial SUMOylation assay to systematically test for SUMO-1 modification of a selection of neuronal proteins containing one or more amino acid sequences predicted as high-probability SUMOylation sites in computer based searches. Of the 39 proteins analysed only 14 sites were posttranslationally modified by SUMO-1, including the group III metabotropic glutamate receptors and the kainate receptor subunit GluR7. These results identify new candidate proteins that may be involved in the SUMO regulation of synaptic activity and also demonstrate that the presence of the Psi-K-x-D/E motif is not sufficient to indicate that a protein can be SUMOylated in this bacterial system. PMID- 18400392 TI - Steering behaviour can be modulated by different optic flows during walking. AB - Optic flow is a typical pattern of visual motion that can be used to control locomotion. While the ability to discriminate translational or rotational optic flows have been extensively studied, how these flows control steering during locomotion is not known. The goal of this study was to compare the steering behaviour of subjects subjected to rotational, translational, or combined (rotational added to translational) optic flows with a focus of expansion (FOE) located to the right, left, or straight ahead. Ten healthy young subjects were instructed to walk straight in a virtual room viewed through a helmet mounted display while the location of the FOE was randomly offset. Horizontal trajectory of the body's centre of mass (CoM), as well as rotations of the head, trunk and foot were recorded in coordinates of both the physical and virtual worlds. Results show that subjects experienced a mediolateral shift in CoM opposite to the FOE location, with larger corrections being observed at more eccentric FOE locations. Head and body segment reorientations were only observed for optic flows containing a rotational component. CoM trajectory corrections in the physical world were also of small magnitude, leading to deviation errors in the virtual world. Altogether, these results suggest a profound influence of vision, especially due to the pattern of visual motion, on steering behaviours during locomotion. PMID- 18400393 TI - Effects of environmental enrichment and social isolation on sucrose consumption and preference: associations with depressive-like behavior and ventral striatum dopamine. AB - Little attention has been directed towards environmental control of sensitivity to natural reward and its possible relationship with other motivated behaviors, besides the well-known effects of environmental enrichment and social isolation on drug self-administration and locomotor sensitization to psychostimulants. Here, we investigate the effects of these rearing conditions on sucrose consumption and preference, and tissue levels of striatal dopamine. The possible relationship among sucrose intake, immobility behavior in the forced swimming test, and dopamine concentration was explored through correlation and regression analyses. Even though all animals preferred sucrose over water, we found, that during postnatal period, isolated rats consumed more sucrose than control or enriched littermates. In isolated rats sucrose intake correlated positively with ventral but not with dorsal striatum dopamine, even when striatal dopamine did not differ among groups. Especially in isolated animals immobility behavior was positively predicted by differences in sucrose intake. The dopamine concentration did not correlate with immobility behavior. Taken together, the present data support previous findings regarding the effects of early life events upon reward sensitivity and depressive-like behavior, and also provide further evidence about the relationship between these motivated behaviors and the likely role of ventral striatum dopamine in regulating them. PMID- 18400394 TI - Early cytokine gene expression in mouse CNS after peripheral nerve lesion. AB - The generation and maintenance of pain after peripheral nerve injury are thought to be influenced by cytokine signaling. Chronic constriction injury (CCI) to mouse sciatic nerve leads to an early local upregulation of pro-inflammatory cytokines already 1 h after the lesion. The early regulation of cytokines in pain related CNS areas is largely unknown. We investigated cytokine regulation in the lumbar spinal cord, hypothalamus, thalamus, hippocampus, and frontal cortex in C57Bl/6J mice after lesioning the right sciatic nerve by CCI. The gene expression of tumor necrosis factor-alpha (TNF), interleukin (IL)-1beta, IL-4, and IL-10 was analyzed by quantitative real-time-PCR from 1 to 12 h after surgery or until values were back to baseline. CCI led to an early downregulation of TNF and IL 1beta mRNA in distinct brain areas and in the lumbar spinal cord with a maximum decrease within the first 6h after CCI. The reduction of TNF mRNA was inhibited by the NMDA receptor antagonist (+)-MK-801, while the calpain inhibitor MDL-28170 had no effect. Our results suggest an early cytokine regulation in the CNS after peripheral nerve lesion, which is opposite in direction to that in the periphery and which is partly mediated by the NMDA receptor system. PMID- 18400395 TI - Motor activity induces release of serotonin in the dorsal horn of the rat lumbar spinal cord. AB - Literature highlights that serotonergic descending pathways are implicated in somatosensory functions in the spinal cord and that serotonin (5-HT) in the dorsal horn might play a role in motor function through proprioceptive feedback. We hypothesized that 5-HT release in dorsal horn might represent an important factor in the completion of locomotion by facilitation of the spinocerebellar tract and/or by modulation of spinal reflex pathways. The present study demonstrates that during locomotor activity, 5-HT is released in layers II, III, IV, V of Rexed. Microdialysis in combination with HPLC was used to measure concentrations of neurotransmitters in the lumbar dorsal horn before, during, and after a treadmill running exercise. Our results show a significant 41% increase of 5-HT release within the dorsal horn during the exercise. 5-HT release is temporally related to exercise. The present study demonstrates that dorsal horn 5 HT release might modulate locomotion. PMID- 18400396 TI - Variations in cortical thickness with dementia severity in Alzheimer's disease. AB - Previous magnetic resonance imaging (MRI) studies have used volumetric methods to investigate cerebral atrophy and showed its linear pattern with the measure of dementia severity in Alzheimer's disease (AD). This study analyzed the phase- and region-specific changes in cortical thickness with dementia severity. In 43 normal controls and 60 AD patients with clinical dementia rating (CDR) (0.5, n=21; 1, n=28; 2, n=11), the cortical thickness was measured using automated surface-based analysis of MRI data. Statistical analyses were performed to investigate overall the hemispheric mean thicknesses as well as the topography of cortical atrophy based on vertices in the groups. No significant difference in cortical thickness was observed for the mild (from CDR=0.5 to 1) stage of dementia. In contrast, a significant reduction of cortical thickness occurred from CDR=1 to 2. Topographic analysis of cortical atrophy showed that the significant cortical thinning in CDR=0.5 relative to normal was found in most association cortices, with this being more extensive than previously reported. There were significant cortical atrophies between CDR=1 and 2 in the frontal, inferolateral temporal, inferior parietal lobule, medial occipital, and posterior cingulated regions. Our results confirm and extend previous findings, suggesting that widespread cortical thinning occurs before the onset of dementia (from normal to CDR=0.5), and that once dementia starts, cortical atrophy in association cortices accelerates in moderate AD (from CDR=1 to 2). PMID- 18400397 TI - Changes in BOLD transients with visual stimuli across 1-44 Hz. AB - The dependency of positive BOLD (PBOLD) and post-stimulus undershoot (PSU) on the temporal frequency of visual stimulation was investigated using stimulation frequencies between 1 and 44 Hz. The PBOLD peak at 8 Hz in primary visual cortex was in line with previous neuroimaging studies. In addition to the 8 Hz peak, secondary peaks were observed for stimulation frequencies at 16 and 24 Hz. These additional local peaks were contrary to earlier fMRI studies which reported either a decrease or a plateau for frequencies above 8 Hz but in line with electrophysiological results obtained in animal local field potential (LFP) measurements and human steady-state visual evoked potential (SSVEP) recordings. Our results also indicate that the dependency of PSU amplitude on stimulus frequency deviates from that of PBOLD. Although their amplitudes were correlated within the 1-13 Hz range, they changed independently at stimulation frequencies between 13 and 44 Hz. The different dependency profiles of PBOLD and PSU to stimulation frequency points to different underlying neurovascular mechanisms responsible for the generation of these BOLD transients with regard to their relation to inhibitory and excitatory neuronal activity. PMID- 18400398 TI - Subthreshold noise facilitates the detection and discrimination of visual signals. AB - It has recently been shown that noise can facilitate signal detection in the sensory systems. We have previously shown that contrast detection sensitivity in the visual perception is improved in the presence of noise by measuring the detection threshold (Sasaki et al. Effect of noise on the contrast detection threshold in visual perception, Neurosci. Lett. 408 (2006) 94-97). In the present study, we reexamined the previous findings by measuring the percent correct detection rates in a two-alternative forced-choice (2AFC) paradigm and examined further whether the addition of noise improved the discrimination of the visual patterns. In the first experiment, the addition of weak noise facilitated the detection rate, thus confirming the previous findings. Furthermore, a significant correlation between the control detection rate without noise and the noise intensity that induced the maximum facilitation of the detection rate was found. In the second experiment, the addition of a subthreshold noise facilitated the discrimination of visual patterns. These results showed that the effect of noise that induces facilitation in signal detection is robust regardless of the methods used for measuring the detection sensitivity, and that pattern discrimination can also be improved by the addition of noise. PMID- 18400399 TI - Activation of endothelial nitric oxide synthase following spinal cord injury in mice. AB - Endothelial nitric oxide synthase (eNOS) plays a neuroprotective role after cerebral ischemia through the production of NO, which enhances cerebral blood flow. However, precise details regarding activation of eNOS after spinal cord injury (SCI) largely remain to be elucidated. In the present study we investigated chronological alteration and cellular location of eNOS and phosphorylated (p)-eNOS at Ser(1177) following SCI in mice. Western blot analysis showed eNOS to be significantly phosphorylated at Ser(1177) from 1 to 2 days after mild SCI, with gradual decrease thereafter. Immunohistochemistry revealed the p-eNOS to be mainly expressed in the endothelial cells of microvessels within gray matter under these conditions. These findings suggest that mild SCI activates eNOS in the subacute stage, which increases spinal cord blood flow and may be involved in protective and repair responses. PMID- 18400400 TI - Fibromyalgia relapse evaluation and efficacy for durability of meaningful relief (FREEDOM): a 6-month, double-blind, placebo-controlled trial with pregabalin. AB - This was a multicenter, double-blind (DB), placebo-controlled, randomized discontinuation trial to evaluate the efficacy of pregabalin monotherapy for durability of effect on fibromyalgia (FM) pain. The trial included a 6-week open label (OL) pregabalin-treatment period followed by 26-week DB treatment with placebo or pregabalin. Adults with FM and 40-mm score on 100-mm pain visual analog scale (VAS) were eligible. During OL weeks 1-3, patients received escalating dosages of pregabalin to determine their optimal dosages. During OL weeks 4-6, patients received their optimal fixed dosages (300, 450, 600mg/d). To be randomized, patients must have had 50% decrease in pain VAS and a self-rating of "much" or "very much" improved on Patient Global Impression of Change (PGIC) at the end of OL. Double-blind treatment was with placebo or the patient's optimal fixed dosage of pregabalin. Primary outcome was time to loss of therapeutic response (LTR), defined as <30% reduction in pain (from OL baseline) or worsening of FM. A total of 1051 patients entered OL; 287 were randomized to placebo, 279 to pregabalin. Time to LTR was longer for pregabalin versus placebo (P<.0001). Kaplan-Meier estimates of time-to-event showed half the placebo group had LTR by Day 19; half the pregabalin group still had not lost response by trial end. At the end of DB, 174 (61%) placebo patients met LTR criteria versus 90 (32%) pregabalin patients. Pregabalin was well tolerated, though 178 (17%) discontinued during OL for treatment-related adverse events (AE), and more pregabalin than placebo patients discontinued for AEs during DB. In those who respond, pregabalin demonstrated durability of effect for relieving FM pain. PMID- 18400401 TI - Isolation and characterization of a gene encoding carbonic anhydrase from Ostertagia ostertagi and quantitative measurement of expression during in vivo exsheathment. AB - The first event in the establishment of Ostertagia ostertagi infection in cattle is exsheathment. Exsheathment is the process whereby the L(2) cuticle retained from the previous molt is cast from the L3. For those trichostrongyle nematode species with a predilection site in the abomasum, such as O. ostertagi, exsheathment is initiated as the larvae pass through the rumen. Although the stimulus for exsheathment is not known, previously reported biochemical studies suggest a major role for the enzyme carbonic anhydrase (CA). Partial support for this hypothesis comes from the reported failure of the Haemonchus contortus L3 to exsheath following pretreatment with ethoxzolamide, a known inhibitor of CAs. Although convincing, a CA has not been previously reported from a trichostrongylid nematode. Therefore, our objective was to isolate a CA gene from O. ostertagi L3 and begin initial characterization studies. This work resulted in the successful isolation, cloning and sequencing of the first CA isolated from a gastrointestinal nematode. The gene, designated OoCA, shows 90.5% sequence identity with the CA eukaryotic consensus sequence, 78% similarity to the Caenorhabiditis elegans cah-6 and 55% similarity to the human CAIII. Sequence analysis of the genomic DNA encoding OoCA shows 8 exons and 7 introns covering 4.5kb. The first 1758 bases of the promoter region suggest OoCA may be regulated in part by transcription factors associated with hypoxic signaling and development. The mRNA profile of OoCA in exsheathing O. ostertagi L3 suggests this particular CA may play a role in immediate early developmental events following exsheathment initiation. PMID- 18400402 TI - Babesia divergens-like organisms from free-ranging chamois (Rupicapra r. rupicapra) and roe deer (Capreolus c. capreolus) are distinct from B. divergens of cattle origin - an epidemiological and molecular genetic investigation. AB - In 2005 and 2006, three adult female chamois (Rupicapra r. rupicapra) were found dead with signs of acute babesial infection in the eastern Swiss Alps. PCR on DNA extracted from blood or spleen of the carcasses revealed sequence identity of the amplified part of the 18S rRNA gene with GenBank entries attributed to Babesia divergens of cattle origin or B. capreoli of wild ruminant origin which have never been described before in this region. Examination of 424 blood samples from 314 head of cattle from this area by IFAT, microscopy and PCR provided no evidence for babesial infection. Six of 887 ticks collected from cattle were PCR positive, and sequencing revealed Babesia sp. genotype EU1 in five and B. divergens/B. capreoli in one of them. A Babesia isolate of chamois, two isolates of roe deer from the same region and one isolate of a roe deer from the north western Swiss Alps were genetically compared with two Swiss B. divergens isolates of cattle origin by analysing the genomic rDNA locus. Whereas the near full length sequences of the 18S rRNA gene were virtually identical among all six isolates (>99.4% identity), distinct differences between the two isolates from cattle on the one hand and the four isolates from free-ranging ruminants on the other hand were observed in the sequences of the internal transcribed spacers 1 and 2 (ITS1, ITS2) and part of the 28S rRNA gene. These results indicate that, albeit genetically very closely related, these babesial organisms from cattle and from free-ranging ruminants indeed are distinguishable organisms with different host specificities, and they support the use of the discrete species name B. capreoli for the B. divergens-like organisms from chamois and roe deer. PMID- 18400403 TI - Amitriptyline induces nuclear transcription factor-kappaB-dependent glutamate transporter upregulation in chronic morphine-infused rats. AB - We previously showed that intrathecal co-administration of amitriptyline with morphine upregulates the expression of the glial glutamate transporters glutamate aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1) and restores neuronal glutamate transporter excitatory amino acid carrier 1 (EAAC1) expression in chronically morphine-infused rats. The present study examined the role of nuclear transcription factor-kappaB (NF-kappaB) in the regulation of the expression of GLAST, GLT-1, and EAAC1 following long-term amitriptyline/morphine co-infusion. Male Wistar rats were implanted with two intrathecal catheters with or without a microdialysis probe; one of the catheters was used for continuous infusion of saline (control), morphine (15 microg/h), or morphine plus amitriptyline (both 15 microg/h) for 5 days, while the other was used for a single daily intrathecal injection of the NF-kappaB inhibitor Ro106-9920 (10 microl of 10 microM) for 5 days. We found that amitriptyline co-infusion restored the antinociceptive effect of morphine (4.5-fold right-shift in the morphine dose response curve compared with a 65-fold right-shift in its absence) and this effect was inhibited by Ro106-9920 administration (48-fold right-shift). Moreover, amitriptyline/morphine co-infusion increased IkappaBalpha phosphorylation and the translocation of NF-kappaB p65 from the cytosol to the nucleus. Daily intrathecal injection of Ro106-9920 prevented the amitriptyline/morphine-induced NF-kappaB p65 translocation and reversed the amitriptyline/morphine-induced GLAST and GLT-1 upregulation and inhibited the restoration of EAAC1 expression. The Ro106-9920 injections abolished the inhibitory effect of amitriptyline on the morphine-evoked release of excitatory amino acids into the spinal cerebrospinal fluid (CSF) dialysates. In conclusion, amitriptyline/morphine co-infusion restores the antinociceptive effect of morphine and upregulates GLAST and GLT-1 expression and restores EAAC1 expression to baseline levels, thus reducing excitatory amino acid levels in the spinal CSF dialysates. The mechanism involves activation of the NF-kappaB pathway, but may also involve other pathways. PMID- 18400404 TI - Kainate receptors in epilepsy and excitotoxicity. AB - Kainate (KA), an analog of glutamate, is a potent neurotoxin that has long been known to induce behavioral and electrophysiological seizures as well as neuropathological lesions reminiscent of those found in patients with temporal lobe epilepsy. More than a decade after the initial KA studies, molecular cloning of ionotropic glutamate receptors identified a family of receptors that binds KA with high affinity. The present review explores the links between the epileptogenic and excitotoxic actions of KA and the function of kainate receptors (KARs) in the activity of neuronal networks. We first present evidence that KARs are the main targets of KA to produce the epileptogenic and excitotoxic effects of KA and KA analogs, and we discuss the mechanisms of action of KA. Then the review evaluates the involvement of KARs activated by the endogenous agonist glutamate in the generation and propagation of epileptiform activity. Finally, we report recent findings proposing KARs as targets of antiepileptic drugs and neuroprotective agents. PMID- 18400405 TI - Promotion of neurite outgrowth by fibroblast growth factor receptor 1 overexpression and lysosomal inhibition of receptor degradation in pheochromocytoma cells and adult sensory neurons. AB - Basic fibroblast growth factor (FGF-2) is up-regulated in response to a nerve lesion and promotes axonal regeneration by activation of the tyrosine kinase receptor fibroblast growth factor receptor 1 (FGFR1). To determine the effects of elevated FGFR1 levels on neurite outgrowth, overexpression was combined with lysosomal inhibition of receptor degradation. In pheochromocytoma (PC12) cells, FGFR1 overexpression resulted in flattened morphology, increased neurite outgrowth and activation of extracellular signal-regulated kinase (ERK) and AKT. Degradation of FGFR1 was inhibited by the lysosomal inhibitor leupeptin and by the proteasomal inhibitor lactacystin. In rat primary adult neurons, FGFR1 overexpression enhanced FGF-2-induced axon growth which was further increased by co-treatment with leupeptin. Lysosomal inhibition of receptor degradation concomitant with ligand stimulation of neurons overexpressing FGFR1 provides new insight in tyrosine kinase receptor-mediated promotion of axon regeneration and demonstrates that adult sensory neurons express sub-optimal levels of tyrosine kinase receptors for neurotrophic factors. PMID- 18400407 TI - Colocalization of CB1 receptors with L1 and GAP-43 in forebrain white matter regions during fetal rat brain development: evidence for a role of these receptors in axonal growth and guidance. AB - There is recent evidence supporting the notion that the cannabinoid signaling system plays a modulatory role in the regulation of cell proliferation and migration, survival of neural progenitors, neuritic elongation and guidance, and synaptogenesis. This assumption is based on the fact that cannabinoid 1-type receptors (CB(1) receptors) and their ligands emerge early in brain development and are abundantly expressed in certain brain regions that play key roles in these processes. We have recently presented in vivo evidence showing that this modulatory action might be exerted through regulating the synthesis of the cell adhesion molecule L1 that is also a key element for those processes. To further explore this issue, we conducted here immunohistochemical studies aimed at determining the cellular substrates of CB(1) receptor-L1 interactions in the rat brain during late fetal development. In this period, we previously found that the activation of CB(1) receptors increased L1 synthesis in several forebrain white matter regions but not in gray matter areas. Using double labeling studies, we observed here colocalization of both proteins in fiber tracts including the corpus callosum, the adjacent subcortical white matter, the internal capsule and the anterior commissure. Experiments conducted with cultures of fetal rat cortical nerve cells revealed that L1 is present mainly in neurons but not in glial cells. This fact, together with the results obtained in the double labeling studies, would indicate that L1 and CB(1) receptors should possibly be present in axons elongating through these white matter tracts, or, alternatively, in migrating neurons. Further experiments confirmed the presence of CB(1) receptors in elongating axons, since these receptors colocalized with growth-associated protein 43 (GAP-43), a marker of growth cones, but not with synaptophysin, a marker of active synaptic terminals, in the same forebrain white matter regions. Lastly, using cultured fetal rat cortical neurons, we also observed that the activation of cannabinoid receptors increased the levels of the full-length L1 and altered those of some active proteolytic fragments of this protein whose generation has been associated with specific steps in the process of neuritic elongation in cultured neurons. In summary, we have demonstrated that the effects caused by cannabinoid agonists on L1 are facilitated by the colocalization of this cell adhesion molecule with CB(1) receptors in several forebrain white matter regions during fetal brain development. We have provided strong evidence that this phenomenon occurs in axons elongating through these white matter tracts, and we have explored in vitro how cannabinoid receptors influence L1 levels. Considering the role played by L1 in different events related to neural development, our observations support the occurrence of a physiological mechanism by which the cannabinoid system might regulate the process of axonal growth and guidance through regulating the synthesis and function of L1. PMID- 18400406 TI - Dendritic Ca2+ responses in neonatal lateral superior olive neurons elicited by glycinergic/GABAergic synapses and action potentials. AB - During development, GABA/glycinergic connections from the medial nucleus of the trapezoid body (MNTB) to the lateral superior olive (LSO) gradually change from being depolarizing to being hyperpolarizing. Previous studies have shown that depolarizing MNTB-LSO synapses can trigger action potentials and increase the concentration of intracellular calcium. In the present study we used confocal calcium imaging combined with whole-cell patch clamp recordings to investigate how depolarizing MNTB inputs in neonatal rats and mice increase the calcium concentration in the dendrites of LSO neurons. Our results show that subthreshold synaptic responses can elicit local dendritic calcium responses while suprathreshold responses reliably generate global calcium responses that are observed in all dendritic processes. The amplitude of global dendritic calcium responses increased with distance from the soma. Global calcium responses were blocked by tetrodotoxin and could not be recovered by somatic injection of action potential waveforms indicating that global calcium responses are generated by back-propagating sodium action potentials. PMID- 18400408 TI - Different protein profiles in inferior colliculus and cerebellum: a comparative proteomic study. AB - The characteristic features of individual brain regions are determined by anatomical, physiological, and biochemical properties, which are caused by the nature and amount of proteins expressed. Proteomics is a powerful technology for assessing different protein profiles, comparing hundreds of proteins simultaneously. Here we performed a semi-quantitative proteomic analysis of two prominent brain regions in the male adult rat, the inferior colliculus and the cerebellum. Both play important roles in sensorimotor integration but have distinct anatomical and biochemical features. Soluble proteins of mainly cytoplasmic origin were obtained through subcellular fractionation, separated by two-dimensional gel electrophoresis, and identified by matrix-assisted laser desorption/ionization mass spectrometry. Out of 169 annotated and quantified spots, 40 (24%) displayed significant differences in intensity between the two brain regions. Of those, 21 spots (containing 26 proteins) were more intense in the inferior colliculus and 19 spots (containing 25 proteins) in the cerebellum. The inferior colliculus displayed a higher abundance of proteins involved in vesicular trafficking, such as dynamin-1 and cofilin-1. In the cerebellum, Ca2+ binding proteins (calbindin and calretinin) as well as 14-3-3 proteins were more abundant. Both protein groups play a central role in cellular signaling. Finally, several differences occurred among proteins involved in cellular energy metabolism. Our study presents a proof of principle to demonstrate marked heterogeneity of proteins between two brain samples. The heterogeneity is likely associated with functional differences, warranting further histological and physiological analyses. PMID- 18400409 TI - Olfactory ensheathing cells promote proliferation and inhibit neuronal differentiation of neural progenitor cells through activation of Notch signaling. AB - A population of neural progenitor cells (NPCs) has been known to exist in adult spinal cord and migrate toward the lesion regions during spinal cord injury (SCI). Although there are some positive effects of the transplanted olfactory ensheathing cells (OECs) on axonal regeneration in SCI, little is known about the effects and the underlying mechanism of these grafted OECs on NPCs. In this study, we have investigated how soluble factors derived from rat OECs regulate the proliferation and differentiation of rat NPCs. The conditioned medium from cultured OECs showed its ability to promote proliferation and inhibit neuronal differentiation of NPCs. Notch signaling was apparently involved in this process. With the addition of DAPT, which inhibited Notch signaling, the effects of OEC conditioned medium on NPCs were blocked. We thus conclude that diffusible factors released from OECs activate the Notch signaling pathway to stimulate the proliferation and suppress neuronal differentiation of NPCs. These findings reveal the likely limitation of using OECs transplantation for SCI repair. PMID- 18400410 TI - Accumulation of beta-amyloid in the brain microvessels accompanies increased hyperphosphorylated tau proteins following microsphere embolism in aged rats. AB - To define mechanisms underlying neurovascular injury following brain embolism induced neurodegeneration, we investigated temporal and spatial pathological changes in brain microvessels up to 12 weeks after microsphere embolism (ME) induction in aged male rats. Mild ME upregulated endothelial nitric oxide synthase (eNOS) and protein tyrosine nitration in brain microvessels. Strong beta amyloid immunoreactivity coincident with increased eNOS immunoreactivity was observed in microvessels. Immunoblotting of purified brain microvessels revealed that beta-amyloid accumulation significantly increased 1 week after ME induction and remained elevated for 12 weeks. Importantly, beta-amyloid accumulation in brain parenchyma was also observed in areas surrounding injured microvessels at 12 weeks. Levels of Alzheimer's-related hyperphosphorylated tau proteins also concomitantly increased in neurons surrounding regions of beta-amyloid accumulation 12 weeks after ME induction, as did glycogen synthase kinase (GSK3beta) (Tyr-216) phosphorylation. Taken together, ME-induced aberrant eNOS expression and subsequent protein tyrosine nitration in microvessels preceded beta-amyloid accumulation both in microvessels and brain parenchyma, leading to hyperphosphorylation of neuronal tau proteins through GSK3beta activation. PMID- 18400411 TI - Pharmacological characterization and gene expression profiling of an L5/L6 spinal nerve ligation model for neuropathic pain in mice. AB - L5/L6 spinal nerve ligation (SNL) in rodents induces behavioral signs similar to the symptoms of neuropathic pain in humans. L5/L6 SNL in rats has been well characterized so far, but there have been few studies using mice. In this study, we established an L5/L6 SNL model in mice and examined the effects of known antinociceptive drugs in the model. We also analyzed the changes in gene expression in dorsal root ganglions with special reference to those which are known to change in a neuropathic pain state to validate the model. Mechanical allodynia in the ipsilateral side paw was observed beginning on day 1 and lasted for at least 2 months following surgery. Diclofenac showed no significant effect on the mechanical allodynia. Gabapentin and pregabalin completely reversed allodynia, but they also caused a decrease in locomotor activity. Duloxetine caused a partial recovery of the threshold. Mexiletine completely reversed allodynia, but it also caused sedation or motor impairment. Morphine caused a partial recovery of the threshold and hyper-locomotion. This mouse L5/L6 SNL model represents a robust mechanical allodynia, which shows a similar pharmacological response to that reported in rats and human patients with neuropathic pain. The pattern changes in gene expression also resembled those reported in rats. This model will therefore be useful for investigation of the effects of novel antinociceptive compounds and the mechanisms of neuropathic pain. PMID- 18400412 TI - The distribution of nitric oxide synthase in the inferior colliculus of guinea pig. AB - The modulation of neuronal activity by the gas nitric oxide is one of the most novel discoveries in neuroscience. In the auditory pathway, the highest expression of nitric oxide synthase is found in the inferior colliculus (IC), an important center for the convergence of parallel ascending pathways traveling in the brainstem, and descending projections from the auditory cortex. Here we use immunocytochemistry with an antibody for neuronal nitric oxide synthase (nNOS), or NOS Type 1, to map the distribution of nNOS expression in the IC of the guinea pig. The results show that nNOS is differentially expressed by both cell bodies and neuropil across its different subdivisions. The highest levels of neuronal staining are seen in the dorsal and lateral cortices, and the commissural nucleus, making them readily distinguishable from the ventro-lateral part of the central nucleus where nNOS expression in neuropil and somata is minimal. Dorso medially, and caudally, however, the region of nNOS expression extends from the dorsal cortex into the area normally designated as the central nucleus, and nNOS is expressed by neurons characteristic of this subdivision. Our findings support the idea of a gradual transition in cell properties rather than a distinct boundary between the central nucleus and the dorsal cortex. This transition zone may provide a cytoarchitectonic substrate for functional interaction between these two subdivisions. PMID- 18400413 TI - Biological challenge procedures used to study co-occurring nicotine dependence and panic disorder. AB - A wide array of biological challenge procedures - including carbon dioxide inhalation, hyperventilation, and breath holding - have been used to model panic in laboratory settings. Originally used to study developmental processes in panic disorder (PD), these procedures, along with nicotine patch administration and self-administered smoking, have recently been applied to help understand the etiology of co-occurring nicotine dependence and PD. The goals of the present paper are to review studies that have employed biological challenges to study the comorbid condition, identify the advantages and limitations of the various procedures, describe desirable outcome measures for use in biological challenges, and present recommendations for future challenge studies in this field. We argue that biological challenges, though in need of standardization, are useful for studying the development, maintenance, prevention, and treatment of comorbid nicotine dependence and PD. PMID- 18400414 TI - Hypothetical molecular mechanisms by which local iron overload facilitates the development of venous leg ulcers and multiple sclerosis lesions. AB - This paper presents a hypothetical model of role for iron in the development of venous leg ulcers and multiple sclerosis. Elevated concentrations of iron were found in the skin affected by venous hypertension and also in the areas of brain with multiple sclerosis lesions. Individuals with hemochromatosis gene (HFE) mutations: C282Y and H63D, which result in a less efficient transport of iron by macrophages, are characterized by an increased risk for venous leg ulcer and multiple sclerosis. Multiple sclerosis is a T cell-mediated disease, and T cells probably participate in the development of venous ulcers. This deleterious role of ferric ions could be related to the regulation of T cell proliferation and apoptosis. Under normal conditions excessive accumulation of T cells cannot take place, because nitric oxide and interferon-gamma drive these cells toward apoptosis. However, in tissues with a high concentration of iron, T lymphocytes proliferate instead of undergoing apoptosis. This is possible due to the internalization of the INF-gammaR2 chain of the interferon-gamma receptor, the downregulation of inducible nitric oxide synthase expression in macrophages and the inactivation of the active site of caspases. Yet, it should be emphasized that this hypothesis does not claim for the increased concentration of iron as a direct causal factor for the development of venous ulcerations or multiple sclerosis, but rather, iron is a factor that modulates and exaggerates the autoimmune process. Iron chelators, administered systemically or locally, should potentially exhibit therapeutic and prophylactic activity against venous leg ulcers and multiple sclerosis. PMID- 18400415 TI - The concentration of deoxyribonucleic acid in plasma from 73 patients with colorectal cancer and apparent clinical correlations. AB - BACKGROUND: Detection of cell-free plasma DNA has considerable potential as a tool for the diagnosis and assessment of the prognosis of many types of cancer. The aim of the present study was to quantify, by spectrophotometry, the cell-free DNA in plasma samples from patients with colorectal cancer at different stages of the disease and to attempt to correlate the resultant values with the clinical picture. METHODS: We reviewed the medical reports of 73 patients, who had undergone resection of primary colorectal cancer. Samples of blood had been taken from each patient immediately prior to surgery. DNA was extracted from samples of plasma and quantified, by spectrophotometry, after a storage period of no longer than 2 years in 89% of the cases examined. RESULTS: The mean(+/-S.D.) concentration of DNA in plasma samples was 108+/-156 ng/microl. We found a statistically significant correlation between the concentration of DNA and the presence of metastases (mainly liver metastases). CONCLUSION: The detection and quantitation of cell-free DNA in plasma, using this simple technique, might be of clinical value for the surveillance of colon cancer patients and the detection of metastases. PMID- 18400416 TI - Nutrition and breast cancer among sporadic cases and gene mutation carriers: an overview. AB - BACKGROUND: Breast cancer is the second leading cause of cancer-related deaths among women in most industrialized countries. Most breast cancers are considered sporadic, with only 5-10% estimated to be due to inherited susceptibility. The objective of this paper is to provide an overview of the effect of nutrition on breast cancer risk among gene mutation carriers as well as those with sporadic breast cancer. METHODS: The published literature from 1999 to 2007 was reviewed to examine the relationship between nutrition and breast cancer among sporadic cases and gene mutation carriers. RESULTS: Evidence suggests that fruits and vegetables, low-fat dairy products, fish, monounsaturated and polyunsaturated fatty acids, vitamin D, calcium, and phytoestrogens may reduce the risk of breast cancer. However, high intake of meat, poultry, total energy, total fat and saturated fatty acids may play a causative role in this disease. CONCLUSIONS: Diet in breast cancer pathogenesis is a modifiable risk factor on which to focus prevention efforts. Identification of the relationship between nutrition and breast cancer among sporadic cases and gene mutation carriers provides necessary data for breast cancer prevention. PMID- 18400417 TI - Risk factors for advanced sporadic colorectal neoplasia in persons younger than age 50. AB - BACKGROUND: Colorectal cancer (CRC) screening is recommended for average-risk adults beginning at age 50. However, 7% of CRC occurs in persons younger than age 50, a group for which risk factors are not well defined. We sought to determine whether a retrospective case-control study could identify risk factors for sporadic CRC and advanced adenomatous polyps (together known as sporadic colorectal neoplasia [CRN]). METHODS: Using the cancer registry, medical records, and endoscopy and pathology reports from six local hospitals, we identified potentially eligible persons with CRN (cases) or controls who had no neoplasia on colonoscopy between January 1, 2000 and December 31, 2002. Consenting subjects completed a survey encompassing medical and family history, physical measures, lifestyle habits, and diet. RESULTS: Surveys were completed by 20 (15%) of 130 potentially eligible cases and by 54 (13%) of 408 potentially eligible controls. The following factors differed between cases and controls: living with a spouse/significant other (55% vs. 80%; P=0.034); prior pelvic irradiation (20% vs. 2%; P=0.019); having a first-degree relative with CRC (25% vs. 7%; P=0.05); having had a prior sigmoidoscopy, colonoscopy, or barium enema (15% vs. 41%; P=0.038); and lightest weight since age 21 (155lbs vs. 135lbs; gender-adjusted P=0.049). CONCLUSIONS: The low recruitment rate of this retrospective case control study precludes its use for a larger, more definitive study. Several potential risk factors for advanced sporadic CRN were identified. It remains to be determined whether these factors represent an artifact of selection bias or true risk factors that may be used to stratify risk and target screening in persons under age 50. PMID- 18400418 TI - Pro-inflammatory cytokines and prostate-specific antigen in hyperplasia and human prostate cancer. AB - BACKGROUND: The aim of this study was to relate the expression, analyzed by Western blot and immunohistochemistry, of several pro-inflammatory cytokines, including IL-1, IL-6 and TNF-alpha, with serum levels of prostate-specific antigen (PSA) in normal and pathologic (hyperplasia and cancer) prostate tissues to elucidate their possible role in tumor progression. We are also discussing the possible use of these cytokines as a potential therapeutic target. METHODS: The study was carried out in 5 normal, 25 benign prostatic hyperplastic (BPH) and 17 cancerous human prostates (PC). Immunohistochemical and Western blot analysis were performed. Serum levels of PSA were assayed by an immulite autoanalyzer. RESULTS: The most relevant results showed that in BPH, IL-1alpha, IL-6 and tumor necrosis factor (TNF) were only expressed in patients with PSA serum levels of 0 4 or 4-20 ng/ml, but not in the group >20 ng/ml. In PC these cytokines were only expressed in patients with PSA serum levels >4 ng/ml. CONCLUSIONS: In PC there was an association between the high expression of pro-inflammatory cytokines (TNFalpha, IL-6, IL-1), elevated serum levels of PSA and cancer progression. A better understanding of the biologic mechanism of this association may provide new targets for therapy in these patients. PMID- 18400419 TI - A two-stage model for concurrent sequences. AB - Schneider and Davison [Schneider, S.M., Davison, M., 2005. Demarcated response sequences and the generalised matching law. Behav. Proc. 70, 51-61] showed that the generalised matching law applied to concurrent free-operant two-response sequences. When sufficient temporal spacing is required between the responses, however, neither the response-level nor the sequence-level forms of the generalised matching law provide good fits. An alternative "two-stage sensitivity" model with fewer free parameters features two types of sensitivity to the reinforcement contingencies on sequences. When temporal spacing between the responses is long, the "response distribution sensitivity" parameter describes sensitivity only of the individual responses to the sequence-level contingencies. At a threshold level, this sensitivity reaches a maximum. When spacing is shorter than threshold, the "response order sensitivity" parameter reflects a new sensitivity to the order of the responses within sequences. As this sensitivity approaches its maximum, sequence matching is achieved. For both stages, a changeover parameter describes bias against sequences that require changeovers between the two responses. The model fit data ranging from near response matching with long minimum inter-response times (IRTs) to sequence matching with no minimum IRTs, using two species and a variety of sequence reinforcer distributions. Rats differed from pigeons in achieving sequence matching only at a nonzero minimum IRT. In a comparison based on pigeon data with no minimum IRT, the two-stage sensitivity model was more efficient than the generalised matching law according to the Akaike criterion. The logic of the model suggests a new way of understanding the mechanisms underlying behavioural units. PMID- 18400420 TI - SQAB 2007: completing the circle. PMID- 18400421 TI - Comparison of the pathogenicity in three different Korean canine parvovirus 2 (CPV-2) isolates. AB - Canine parvovirus type 2 (CPV-2) is a major pathogen inducing acute hemorrhagic gastroenteritis in dogs. Despite the identification of numerous CPV-2 variants (from CPV-2a to CPV-2c), the pathogenic differences among the CPV-2 variants in dogs have not been evaluated. The aim of this study was to compare the pathogenicity of CPV-2 variants (CPV-2a-I, CPV-2a-V and CPV-2b) isolated mainly from Korea. We evaluated the pathogenicity of three different CPV-2 variants, by performing clinical, hematological, serological and histopathological examinations after experimentally inoculating three types of CPV-2 variants into young puppies. We found that the overall pathogenicity of the CPV-2a variants (CPV-2a-I and 2a-V) was severer compared to the CPV-2b variant. In addition, there was no significant difference in pathogenicity between the two CPV-2a variants. Our findings indicate that there are differences in the pathogenicity of CPV-2 variants in dogs, which may be useful to understand the different pathobiology of the CPV-2 variants. PMID- 18400422 TI - Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein. AB - S1D (residues 636-789) is a neutralizing epitope region on the spike protein (S) of porcine epidemic diarrhea virus (PEDV). To accurately identify epitopes on S1D, the S1-phage library containing the gene encoding the S1D region of PEDV S protein was micropanned by six specific monoclonal antibodies (McAbs) against the S1D region. These micropanned epitope regions (MER) were focused on 696-779 amino acids of the S protein. To further map epitopes of the MER, seven overlapping mini-fragments covering MER nucleotides were separately synthesized and expressed in Escherichia coli BL21 with a GST tag. These mini-GST fusion proteins were scanned by ELISA and Western blotting with the six McAbs, and the result showed that S1D5 (residues 744-759) and S1D6 (residues 756-771) are two linear epitopes of the PEDV S protein. The antisera of the epitopes S1D5 and S1D6 could react with the native S protein of PEDV. Furthermore, Pepscan of the two linear epitopes demonstrated that SS2 ((748)YSNIGVCK(755)) and SS6 ((764)LQDGQVKI(771)) are two core epitopes on S1D5 and S1D6, respectively, located on the S protein of PEDV. PMID- 18400423 TI - Sleep fragmentation and evidence for sleep debt in alcohol-exposed infants. AB - BACKGROUND: Infants exposed prenatally to alcohol are at increased risk for poor neurodevelopmental outcome including Sudden Infant Death Syndrome. AIM: To examine the relationship between prenatal alcohol exposure, sleep, arousal and sleep-related spontaneous motor movements in early infancy. STUDY DESIGN: Low income women (N=13) were interviewed regarding pre- and pregnancy rates of alcohol, cigarette smoking and other substance use in the perinatal period. Infants were examined in a laboratory nap study using EEG, videography and actigraphy at 6-8 weeks of age. Estimates of maternal pre- and pregnancy alcohol use were used to divide infants into high vs. low maternal alcohol use groups. SUBJECTS: Mother-infant dyads recruited from a family practice clinic. OUTCOME MEASURES: Sleep-related spontaneous movements, behavioral state, and maternal assessments of infant alertness and irritability. RESULTS: Pre-pregnancy rates of alcohol consumption including binge drinking correlated with maternal report of poor infant alertness, and increased irritability. High maternal exposure groups showed increased sleep fragmentation, e.g., frequency and duration of wakefulness following sleep onset and decreased active sleep. Bout analysis of the temporal structure of sleep-related spontaneous movements showed significantly reduced bout duration associated with high maternal alcohol use. CONCLUSION: These results present evidence that prenatal alcohol exposure disrupts postnatal sleep organization and suppresses spontaneous movements during sleep, and increased sleep fragmentation promotes sleep deprivation. Results are consistent with the SIDS model of chronic sleep debt and suggest that attenuated sleep-related movements should be examined as an important modulator of cardiorespiratory functions during sleep in high-risk groups. PMID- 18400424 TI - Improved delivery of cromolyn from oral proliposomal beads. AB - Proliposomal bead formulations for improved oral delivery of cromolyn (BCS Class III compound) were formulated. Phospholipid (distearylphosphatidylcholine) cholesterol-surfactant (Tween 80/sodium cholate) systems were spray-coated on beads containing cromolyn sodium and the dosage forms were characterized for vesicle formation and encapsulation efficiency. Delivery of cromolyn sodium from this novel dosage form was evaluated across the Caco-2 and everted rat intestinal sac model. Spontaneous formation of vesicles upon dilution of beads was observed. Enhancement in cromolyn transport was higher with phospholipids-surfactant proliposomal formulations compared to surfactant-free lipid formulations or pure surfactant solutions, most significant enhancement being with formulations with low surfactant concentration. No evidence of cellular damage to Caco-2 monolayers (e.g. significant decrease in the TEER) or change in transport and tissue accumulation of a marker molecule in the intestinal tissue model was observed. This indicated enhancement of transport via transcellular routes and not due to the modulation of the tight junctions or cell disruption. Results suggest that phospholipids-surfactant proliposomal beads offer a good potential for improved oral delivery of cromolyn. PMID- 18400425 TI - pH control of nucleophilic/electrophilic oxidation. AB - Finding formulations that prevent degradation of the active pharmaceutical ingredient is an essential part of drug development. One of the major mechanisms of degradation is oxidation. Oxidative degradation is complex, and can occur via different mechanisms, such as autoxidation, nucleophilic/electrophilic addition, and electron transfer reactions. This paper uses three model compounds and determines the mechanisms of oxidation and strategies to reduce degradation. The mechanism of oxidation was established by comparing the results of different forced degradation experiments (radical initiation and peroxide addition), computational chemistry to those of formulated drug product stability. The model compounds chosen contained both oxidizable amine and sulfide functional groups. Although, both oxidative forced degradation conditions showed different impurity profiles the peroxide results mirrored those of the actual stability results of the drug product. The major degradation pathway of all compounds tested was nucleophilic/electrophilic oxidation of the amine to form N-oxide. Strategies to prevent this oxidation were explored by performing forced degradation experiments of the active pharmaceutical ingredient (API) in solution, in slurries containing standard excipient mixtures, and in solid formulation blends prepared by wet granulation. The reaction was significantly influenced by pH in solvent and excipient slurries, with 100% degradation occurring at basic pH values (>pH 8) and no degradation occurring at pH 2 upon exposure to 0.3% peroxide. Wet granulated blends were also stabilized by lowering the pH during granulation through the addition of citric acid prior to the solution of peroxide, resulting in little (0.02% maximum) or no degradation for the four different blends after 6 week storage at 40 degrees C/75%RH. PMID- 18400426 TI - Evaluation of gatifloxacin penetration into skeletal muscle and lung by microdialysis in rats. AB - This study aimed to investigate gatifloxacin distribution into skeletal muscle and lung interstitial fluid by microdialysis and to correlate free tissue and free plasma levels of the drug. Microdialysis recoveries were determined in vitro by extraction efficiency and retrodialysis at 80, 160 and 400 ng/ml resulting in 33.5+/-1.3%, 33.1+/-1.2%, 31.8+/-2.7% and 31.4+/-2.6%, 33.1+/-2.2%, 30.6+/-3.3%, respectively. In vivo recovery by retrodialysis in Wistar rats' skeletal muscle and lung were 29.1+/-1.0% and 30.7+/-1.4%, respectively. The recovery was constant and independent on the method or media used. Gatifloxacin tissue penetration was investigated after intravenous dosing of 6 mg/kg to Wistar rats. Free skeletal muscle, lung and plasma profiles were virtually superimposable resulting in similar area under the curve (AUC(0-9)) of 3888+/-734 ng h/ml, 4138+/-1071 ng h/ml and 3805+/-577 ng h/ml, respectively (alpha=0.05). The tissue distribution factors were 1.02 and 1.08 for muscle and lung relative to plasma. In conclusion, free plasma levels are a good surrogate for gatifloxacin active levels at the infection site. PMID- 18400427 TI - Fructus Corni suppresses hepatic gluconeogenesis related gene transcription, enhances glucose responsiveness of pancreatic beta-cells, and prevents toxin induced beta-cell death. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Fructus Corni, the fruits of Cornus officinalis Sieb. et Zucc., is one important ingredient in Quei Fu Di Huang Wan, a Chinese herbal mixture. AIM OF THE STUDY: In the present study, additional anti-diabetic actions of Fructus Corni on transcriptional regulation of hepatic gluconeogenesis or beta-cell functions were investigated. MATERIALS AND METHODS: Insulin mimetic action of Fructus Corni on dexamethasone and 8-bromo-cAMP induced phosphoenolpyruvate carboxykinase (PEPCK) expression in H4IIE cells was investigated. Besides, BRIN-BD11 cells were used to evaluate both insulinotropic and beta-cell protective effect of Fructus Corni. RESULTS: Firstly, both methanol extract (CO-W-M) and fraction (CO-W-M2) had potent insulin mimic activity on PEPCK expression. Secondly, possibility of both loganin and ursolic acid as the responsible compounds was excluded. Moreover, indication of the existence of phenolic compounds in CO-W-M2 was noticed. In the presence of CO-W-M2, not only was the viability of BRIN-BD11 cells treated with alloxan, streptozotcin, or cytokine mix all significantly increased but also glucose-stimulated insulin secretion was potentiated. CONCLUSIONS: The ability of CO-W-M2 to reduce gene expression for hepatic gluconeogenesis, to protect beta-cell against toxic challenge, and to enhance insulin secretion strengthen the role of Fructus Corni in diabetes therapy. PMID- 18400428 TI - Anticomplementary principles of a Chinese multiherb remedy for the treatment and prevention of SARS. AB - AIM OF THE STUDY: To elucidate the anticomplementary principles of a Chinese multiherb remedy used for the treatment and prevention of SARS, which contains five ingredients of Herba Houttuyniae, Flos Chrysanthemi Indici, Herba Artemisiae Scopariae, Herba Eupatorii and Fructus Tsaoko. MATERIALS AND METHODS: The anticomplementary activity was evaluated from hemolytic assays through the classical pathway (CP) and the alternative pathway (AP) of complement system in vitro. Compounds were isolated using bioactivity-guided fractionation and tested in vitro for their complement-inhibiting properties on the CP and AP. HPLC-DAD ESI-MS was used to assign the isolated compounds in the five ingredient herbs. RESULTS: 15 compounds, including chlorogenic acid (1), rutin (2), hyperoside (3), p-hydroxyacephenone (4), scopoletin (5), quercitrin (6) (3R,4R,6S)-3,6-dihydroxy 1-menthene (7), acaciin (8), scoparone (9), luteolin (10), quercetin (11), apigenin (12), acacetin (13), aristolactam (14), and apigenin-7,4'-dimethyl ether (15) were isolated and identified. Nine flavonoids (2, 3, 6, 8, 10-13, 15) were found to show inhibitory effects towards the CP and AP of complement system, luteolin (10) was the most potent with the CH50 and AP50 values of 0.19 and 0.17 mM. The bioactive flavonoids were mainly derived from Herba Houttuyniae, Flos Chrysanthemi Indici and Herba Artemisiae Scopariae. CONCLUSIONS: A Chinese multiherb remedy used for the treatment and prevention of SARS has robust inhibitory effect on complement system through the CP and AP, and its anticomplementary activity could be attributed to the flavonoids present in some ingredient herbs. Luteolin could be a potential anticomplementary agent. PMID- 18400430 TI - Can the aggregation be a new approach for understanding the mechanism of Traditional Chinese Medicine? AB - "Frequent hitter" phenomenon emerged in the high-throughput screening; one of the most common mechanisms behind artifactual inhibition is that some organic molecules formed large colloid-like aggregates which are able to sequester and thereby inhibit enzymes. To investigate the situation in Traditional Chinese Medicine (TCM), 60 medicinal herbs and 24 Chinese herbal formulae were detected by dynamic light scattering (DLS), and aggregates were observed in all the 84 solution mixtures. The aggregates of two Chinese herbal formulae, 'Xue-Fu-Zhu-Yu Tang' (XF) and 'Jing-Guan Tang' (JG), were not only able to survive in the gastro intestinal environment, but also had the ability to pass through the monolayer of the Caco-2 cell. The activities of XF and JG against three cardiovascular targets were also aggregates-related. Based on these findings, a new possible mechanism of the action of Chinese medicine was proposed. PMID- 18400429 TI - BYHWD rescues axotomized neurons and promotes functional recovery after spinal cord injury in rats. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Buyang Huanwu Decoction (BYHWD), a Chinese prescription that has been used for hundreds of years to treat paralysis, has gained attention recently due to its significant neuroprotective properties. AIM OF THE STUDY: This study was to investigate whether BYHWD treatment protected axotomized rubrospinal neurons (RN) following spinal cord injury (SCI) in rats. MATERIALS AND METHODS: Adult rats received a right lateral funiculus transection at the level between C3 and C4, and were either treated with BYHWD or with distilled water (DW) via gastrogavage. Effects on RN viability and atrophy were determined by Nissl staining, axon regeneration was examined by biotinylated dextran amine (BDA) tracing techniques and functional recovery was studied by a test of forelimb usage during spontaneous vertical exploration. RESULTS: RN cell number and mean somal size were 20% and 29% higher, respectively, in BYHWD treated rats relative to DW-treated rats. There were a small number of BDA labeled axons in the caudal of injury site in BYHWD-treated rats, whereas no caudal axonal regeneration was detected in DW-treated rats. BYHWD-treated rats used the injured forelimb more often than rats treated with DW. CONCLUSIONS: These results indicate that administration of BYHWD following SCI protects injured neurons, promotes regeneration, and enhances functional recovery. PMID- 18400431 TI - Petroleum ether extract of Cnidium monnieri ameliorated scopolamine-induced amnesia through adrenal gland-mediated mechanism in male rats. AB - AIM: Our previous study indicated petroleum ether layer of Cnidium monnieri L. Cuss. (CM) and its ingredient osthole could alleviate scopolamine-induced amnesia in female rats. MATERIALS AND METHODS: Hence, this study was desired to investigate the mechanism of the ameliorating effects of petroleum ether layer of CM on the performance impairment of inhibitory avoidance task and Morris water maze induced by scopolamine in male rats. RESULTS: CM at 0.1-0.6g/kg orally administered 60 min before the training trial ameliorated the scopolamine-induced performance impairment on inhibitory avoidance learning and water maze in male rats. Only adrenalectomy but not peripheral cholinergic antagonist scopolamine methylbromide and catecholaminergic neurotoxin 6-hydroxydopamine blocked the ameliorating effects of CM on scopolamine-induced performance impairment in rats. CONCLUSION: Therefore, we demonstrated that the ameliorating effects of CM on scopolamine-induced performance impairment may be related to activating the adrenal gland and central acetylcholingeric neuron, instead of peripheral nervous system. PMID- 18400432 TI - Comparison of the validity of three dental methods for the estimation of age at death. AB - The aim of the present study was to compare the accuracy, precision, and bias of two macroscopic and one histological age at death estimation methods on human teeth. The sample was comprised of 67 permanent teeth, obtained from 37 individuals aged 20-91 years. Age was predicted according to the methods proposed by Lamendin et al. (LAM), Bang and Ramm (BR), and the quantification of tooth cementum annulations (TCA). TCA was found to be most accurate in all age groups. Its mean absolute error of the estimated age was about half as high as the mean absolute error for both LAM and BR. BR achieved approximately the same mean absolute error as TCA for old adults only. LAM displayed the highest precision in the young and the old age group whereas TCA was more precise in the middle age group. TCA was found to be the most precise method when the precision was calculated for all ages. Considering the bias, all methods displayed a tendency to overestimate age in young and to underestimate it in old specimens. The exception to this rule was TCA, which provided unbiased estimates for young adults. The higher accuracy and precision recommends favouring TCA over LAM and BR, provided that the required know-how and equipment are available. PMID- 18400433 TI - [Recurrent adenomas in ileal pouch following ileoanal anastomosis for familial adenomatous polyposis]. AB - Polyadenoma occuring on ileal pouch is a rare event. The treatment and the frequency of the endoscopic examinations is still matter of controversies. The authors report the case of recurrent endoileal pouch adenoma. PMID- 18400434 TI - [Pseudomalignant endoscopic appearance in hyperplastic polyp at esophagogastric junction: case report]. AB - Hyperplastic polyps are by far the most common benign tumors in the stomach larger than 5mm. Although the malignant potential of hyperplastic gastric polyps was originally denied, a low risk for carcinomatous conversion is now recognized. Its has been recommended that all gastric polyps of 5mm or more in diameter have to be removed. We present here the case of a large hyperplastic gastric polyp located at the esophagogastric junction with endoscopic malignant appearance. The description of its endoscopic and histological characteristics gives us the opportunity to discuss the difficulty of endoscopic diagnosis in the case of gastric polyps and to remind their prognosis. PMID- 18400435 TI - Access to electronic hepatitis C medical files by general practitioners. AB - OBJECTIVES: Hepatitis C virus (HCV) infection is a recognized public health issue in France. Institutional networks were created to improve healthcare practices and facilitate multi-disciplinary care for chronic diseases. The electronic medical file is one of the tools used by the networks to optimize patient care. METHODS: The main objective of this study was to determine what proportion of general practitioners in the Languedoc-Roussillon region of southern France would be interested in using the electronic medical files for patients with HCV infection. A random sample of 20% of the general practitioners in the region was selected and stratified by administrative district of practice. Among them, a telephone survey identified those interested in the hepatitis electronic medical files and following patients with hepatitis C. A more detailed questionnaire was sent to these interested physicians in order to obtain further information. RESULTS: Thirty-seven percent of the general practitioners concerned by the question followed patients with HCV infection. The advantages and disadvantages the physicians associated with use of these files were mostly related to the physician's age, attendance of continuing education courses and internet access. CONCLUSION: This study highlighted the fact that a significant number of general practitioners would be interested in accessing electronic medical files for patients with HCV infection. Considering these findings it might be useful to propose a working group including general practitioners and specialists in order to develop a concrete project for implementing electronic medical files for patients with HCV infection. PMID- 18400436 TI - Budd-Chiari syndrome associated with Behcet's disease. AB - OBJECTIVES: Budd-Chiari syndrome is a rare and serious complication of Behcet's disease, and is the result of occlusion of the major hepatic veins, the adjacent inferior vena cava, or both. The aim of this study was to determine the prevalence, clinical and laboratory findings, and treatment and clinical course of Budd-Chiari syndrome associated with Behcet's disease. METHODS: We analyzed retrospectively the charts of 220 patients fulfilling the international diagnostic criteria of Behcet's disease. From them, we selected those with Budd Chiari syndrome, and analyzed their epidemiological and clinical imaging features and outcomes. RESULTS: Seven male patients, mean age 29 years and already diagnosed with Behcet's disease, had Budd-Chiari syndrome. The clinical course was from subacute to chronic in all cases. Thrombosis of hepatic veins was associated with inferior vena cava thrombosis in six cases. Four patients had other venous thromboses (superior vena cava and lower limbs) and one also had pulmonary emboli. One patient was positive for anticardiolipin antibodies. All patients had anticoagulation therapy, and six had high-dose corticotherapy associated, in two cases, with monthly cyclophosphamid intravenous pulses. Clinical outcome was favourable in six cases, and one patient died of hepatic failure. CONCLUSION: The prevalence of Budd-Chiari syndrome in patients with Behcet's disease is 3.2%, confirming that this syndrome is not uncommon in Behcet's patients. The inferior vena cava is frequently involved in combination with hepatic veins and often associated with other venous thrombosis. The prognosis may be favorable with medical interventions, including anticoagulation, treatment of the vasculitis and the use of diuretics when required. PMID- 18400437 TI - [Esberiven Fort-associated lymphocytic colitis]. PMID- 18400438 TI - [Lymphoepithelial cyst of the pancreas: a new case report]. AB - We report the case of a lymphoepithelial cyst of the pancreas discovered by chance on imaging in a 54-year old man. CT-scan showed a 10 cm hypodense multilocular cystic tumor of the pancreatic isthmus. Fine-needle aspiration did not provide further information. Due to the lack of preoperative diagnosis and mostly because it was not known if the cyst was malignant or benign, the patient underwent a cephalic duodenopancreatectomy. Lymphoepithelial cyst of the pancreas is a rare benign lesion which is difficult to diagnose before surgery. Histologically, the cyst wall is lined by mature keratinizing squamous epithelium and a distinct surrounding lymphoid tissue layer. The cysts are filled with keratin plugs that are not always visualized on imaging. Cytological and histological analysis of fine-needle aspiration material if the sample material is sufficient may help avoid extensive surgery. PMID- 18400439 TI - [Stalevo-associated lymphocytic colitis]. PMID- 18400440 TI - Severe acute pancreatitis related to the use of adefovir in a liver transplant recipient. PMID- 18400441 TI - A case of isolated splenic infarction associated with a foramen ovale and an interatrial septal aneurysm. PMID- 18400442 TI - Radiology in suspected non-accidental injury: theory and practice in The Netherlands. AB - INTRODUCTION: This study evaluates radiological imaging in suspected non accidental injury (NAI) in children below the age of 2 years in the Netherlands. MATERIAL AND METHODS: The study consisted of two parts; first an on-line questionnaire on suspected NAI, amongst radiological practices within the Netherlands. The second part of the study was a retrospective analysis of skeletal surveys in children under the age of 2 years, which were reviewed in an expert centre of forensic medicine on request of the public prosecutor. RESULTS: Out of 116 hospitals 45 (39%) radiologists completed the on-line questionnaire; 8 (8%) of the proposed skeletal surveys complied with the ACR criteria. A total of 29 skeletal surveys in 26 children were reviewed. The median age at the time of the radiographic exam was 3 months for both boys and girls. Only 2 (7%) studies complied with the ACR criteria. DISCUSSION: The results of our study show that, in theory as well as in practice, Dutch radiological practices show a large variation in imaging protocols for suspected NAI. PMID- 18400443 TI - Multidetector spiral CT arthrography of the shoulder. Clinical applications and limits, with MR arthrography and arthroscopic correlations. AB - Although MR imaging and MR arthrography are the first choice modalities for shoulder imaging, CT arthrography (CTA) may be used successfully to address many clinical questions. The advent of submillimeter multiple detector CT technology and subsequent excellent three-plane resolution has considerably increased the quality of CTA examinations and has propelled this technique to the forefront in a growing number of indications. The combined use of iodinated contrast material for fluoroscopic confirmation of the articular position of the needle before injection of gadolinium chelates for MR arthrography offers the unique opportunity to compare CTA and MRA findings in carefully selected cases. This paper illustrates capabilities and limits of CTA for the study of rotator cuff tears, shoulder instability, cartilage lesions, anatomical variants and abnormalities of the glenoid labrum, with correlations to MR arthrography and surgical findings. PMID- 18400444 TI - UV-vis and FTIR-ATR characterization of 9-fluorenon-2-carboxyester/(2 hydroxypropyl)-beta-cyclodextrin inclusion complex. AB - In this work, the usefulness of (2-hydroxypropyl)-beta-cyclodextrin (HP-beta-CyD) as a tool to form an inclusion complex with 9-fluorenonic derivative (AG11) has been investigated, in pure water, by UV absorption. Phase-solubility diagrams allowed the determination of the association constant between AG11 and HP-beta CyD. At the same time, solid binary systems between AG11 and HP-beta-CyD have been prepared in 1:1 stoichiometry by co-precipitation method. In order to confirm the complexation, FTIR spectroscopy in ATR geometry measurements have been performed and the results have been compared with the free compounds and the corresponding physical mixture in the same molar ratio. The nature of the interactions between AG11 and HP-beta-CyD has been elucidated also by applying mathematical procedures such as deconvolution and curve fitting. Improvement of the aqueous solubility is expected to improve the bioavailability of the drug in oral administration. PMID- 18400446 TI - The practical application of implementing the equimolar response principle of chemiluminescent nitrogen detection in pharmaceutical analysis. AB - Since nearly 90% of all developmental and marketed drugs contain at least one nitrogen atom, the implementation of chemiluminescent nitrogen detection (CLND) in pharmaceutical analysis is intriguing due to its equimolar response for nitrogen. Although the documented accuracy of CLND when using a surrogate nitrogen-containing standard may be inadequate for purity and potency determinations, it is acceptable for the quantitation of low-level impurities. A comparison of the quantitative results obtained using both CLND and UV detection for a developmental drug and its related impurities is presented. The results indicated that the impurities can be accurately quantified using a surrogate standard, based on the equimolar response principle of CLND, when the concentrations of the surrogate standard and the component of interest are similar. When the impurities are present at much lower levels than the surrogate standard however, the common practice of direct conversion of area percent to weight percent can result in significant errors using CLND, due to the limited linear dynamic range for CLND. To increase quantitation accuracy, the authors propose that a secondary dilution of the surrogate reference standard solution should be used for the quantitation of low-level impurities. The practical application of this approach for impurity quantitation or as a means to determine impurity relative response factors for use with a traditional UV based method is discussed. PMID- 18400445 TI - HPLC analysis of the antidepressant trazodone and its main metabolite m-CPP in human plasma. AB - The present paper deals with the development of a rapid and feasible high performance liquid chromatographic method for the determination of trazodone and its main active metabolite 3-(1-clorophenyl)piperazine (m-CPP) in human plasma. Trazodone is a second-generation antidepressant with serotonin antagonist activity. The metabolite seems to be involved in the onset of some side effects of trazodone therapy, thus its determination is very important during therapeutic drug monitoring. Separation was achieved using a C8 reversed-phase column and a mobile phase composed of aqueous phosphate buffer (70%), containing triethylamine, at pH 3.5 and acetonitrile (30%). The UV detector was set at 255 nm and loxapine was used as the internal standard. An original pre-treatment procedure of plasma samples was developed, based on solid-phase extraction with C8 reversed phase cartridges (50mg, 1 mL). The obtained extraction yields values were higher than 90% and precision, expressed as R.S.D., was lower than 5.6%. The method was successfully applied to plasma samples from depressed patients undergoing therapy with trazodone; accuracy results were satisfactory (recovery >91%). Thus, the method seems to be suitable for the therapeutic drug monitoring of trazodone and its main active metabolite in depressed patients' plasma. PMID- 18400447 TI - Follicle-stimulating hormone isoforms and plasma concentrations of estradiol and inhibin A in dairy cows with ovulatory and non-ovulatory follicles during the first postpartum follicle wave. AB - Following parturition, all cows display a wave of ovarian follicular growth, but a large proportion fail to generate a preovulatory rise in estradiol, and hence fail to ovulate. Follicle-stimulating hormone (FSH) exists as multiple isoforms in the circulation depending on the type and extent of glycosylation, and this has pronounced effects on its biological properties. This study examined differences in plasma FSH, estradiol, and inhibin A concentrations, and the distribution of FSH isoforms in cows with ovulatory or atretic dominant follicles during the first postpartum follicle wave. Plasma FSH isoform distribution was examined in both groups during the period of final development of the dominant follicle by liquid phase isoelectric focusing. Cows with an ovulatory follicle had higher circulating estradiol and inhibin A concentrations, and lower plasma FSH concentrations. The distribution of FSH isoforms displayed a marked shift toward the less acidic isoforms in cows with ovulatory follicles. A higher proportion of the FSH isoforms had a pI>5.0 in cows with ovulatory follicles compared to those with atretic follicles. In addition, cows with ovulatory follicles had greater dry matter intake, superior energy balance, elevated circulating concentrations of insulin and insulin-like growth factor-I, and lower plasma nonesterified fatty acids. The shift in FSH isoforms toward a greater abundance of the less acidic isoforms appears to be a key component in determining the capability for producing a preovulatory rise in estradiol, and this shift in FSH isoforms was associated with more favorable bioenergetic and metabolic status. PMID- 18400448 TI - Evaluation of immune system function in neonatal pigs born vaginally or by Cesarean section. AB - Full term crossbred sows were selected to study the interaction of the immune system, hypothalamus-pituitary-adrenal axis, and growth in pigs born by Cesarean section (c-section; n=4 sows) or vaginal birth (n=4 sows). Gestation length and birth weight did not differ between vaginal birth and c-section pigs (P=0.34 and 0.62, respectively). Blood and tissue samples were collected from 44 pigs at birth. Forty-five pigs were weaned at 13 d. On d 14, pigs received an i.p. injection of lipopolysaccaride (LPS; 150 microg/kg) or saline at min 0, and blood samples were collected at -20, -10, 0, 5, 10, 20, 40, 60, 90, and 120 min. Vaginal birth pigs had 21% greater average daily gain than c-section pigs on d 14 (P<0.01). Basal serum concentrations of adrenocorticotrophin (ACTH) and cortisol were greater in c-section than vaginal birth pigs at birth (P<0.01) but were not different at 14 d (P=0.99 and 0.80, respectively). LPS increased serum concentrations of ACTH, cortisol, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha; P<0.01) but the response was not different between c-section and vaginal birth (P>0.22). Basal serum concentrations of TNF alpha tended to be greater in c-section vs vaginal birth pigs at 14 d (P=0.0967); however, basal serum concentrations of IFN-gamma tended to be lower in c-section pigs vs vaginal birth pigs at 14 d (P=0.0787). Expression of interleukin (IL)-6, IL-6 receptor, IL-1beta, and TNF-alpha mRNA did not differ between vaginal birth and c-section pigs but changed in an age and tissue dependent manner. Thus, reduced growth rate of c-section pigs is associated with altered immune system function. PMID- 18400449 TI - Functional and morphological changes in the adenohypophysis of dogs with induced primary hypothyroidism: loss of TSH hypersecretion, hypersomatotropism, hypoprolactinemia, and pituitary enlargement with transdifferentiation. AB - From case studies in humans it is known that primary hypothyroidism (PH) may be associated with morphological and functional changes of the pituitary. There is no insight into the time scale of these changes. In this study, seven beagle dogs were followed up for 3 years after the induction of primary hypothyroidism. Three of these dogs were followed up for another 1.5 years while receiving l-thyroxine. Adenohypophyseal function was investigated at 2-month intervals with the combined intravenous injection of CRH, GHRH, GnRH, and TRH, and measurement of the plasma concentrations of ACTH, GH, LH, PRL, and TSH. In addition, after 2 years of hypothyroidism a single TRH-stimulation test and a somatostatin test were performed, with measurements of the same pituitary hormones. Every 6 months the pituitary gland was visualized by computed tomography (CT). Induction of PH led to high plasma TSH concentrations for a few months, where after concentrations gradually declined to values no longer significantly different from pre-PH values. A blunted response to stimulation of TSH release preceded this decline. Basal plasma GH concentrations increased during PH and there was a paradoxical hyperresponsiveness to TRH stimulation. Basal GH concentrations remained elevated and returned only to low values during l-thyroxine treatment. Basal PRL concentrations decreased significantly during PH and normalized after several months of l-thyroxine treatment. The pituitary gland became enlarged in all dogs. Histomorphology and immunohistochemical studies in 4 dogs, after 3 years of PH, revealed thyrotroph hyperplasia, large vacuolated thyroid deficiency cells, and decreased numbers of mammotrophs. Several cells stained for both GH and TSH. In conclusion, with time PH led to a loss of the TSH response to low T4 concentrations, hypersecretion of GH, and hyposecretion of PRL. The enlarged pituitaries were characterized by thyrotroph hyperplasia, large vacuolated thyroid deficiency cells, and double-staining cells, which are indicative of transdifferentiation. PMID- 18400450 TI - Risk drinking behavior among psychotropic drug users in an aging Finnish population: the FinDrink study. AB - Psychotropic drug use and alcohol consumption is increasing among aging Finns. Alcohol use is not recommended with benzodiazepines and some other psychotropic medicines. Concomitant use may lead to accidents and other serious consequences. The aim of this study was to analyze the drinking behavior of psychotropic drug users in an aging Finnish population. This study is part of the ongoing epidemiologic FinDrink study. Self-reported data on alcohol consumption and psychotropic drug use were collected from the Kuopio Ischaemic Heart Disease Risk Factor Study examinations conducted in 1998-2001. Overall, 854 men and 920 women participated in the study. A total of 204 (11.5%) individuals used psychotropic drugs regularly (14.2% of women and 8.5% of men; P<.001). Three quarters of the study population had used alcohol weekly during the preceding year (68.9% of women and 87.5% of men; P<.001). Men who use anxiolytics and sedatives were more likely to drink alcohol at least twice a week (odds ratio=2.42; 95% confidence interval=1.30-4.51), to be binge drinkers (odds ratio=1.86; 95% confidence interval=1.01-3.43) and to be heavy alcohol consumers (odds ratio=2.22; 95% confidence interval= 1.13-4.39) than men not using psychotropics. In women, alcohol consumption and drinking patterns were same between the groups. Our results indicate the potential for alcohol-related health risks among aging Finnish men and women using psychotropic drugs. PMID- 18400451 TI - Effects of alcohol and saccharin deprivations on concurrent ethanol and saccharin operant self-administration by alcohol-preferring (P) rats. AB - Consumption of sweet solutions has been associated with a reduction in withdrawal symptoms and alcohol craving in humans. The objective of the present study was to determine the effects of ethanol and saccharin (SACC) deprivations on operant oral self-administration. Alcohol-preferring (P) rats were allowed to lever press concurrently self-administer ethanol (15% vol/vol) and SACC (0.0125% g/vol) for 8 weeks. Rats were then maintained on daily operant access (nondeprived), deprived of both fluids (2 weeks), deprived of SACC and given 2 ml of ethanol daily, or deprived of ethanol and given 2 ml of SACC daily. All groups were then given 2 weeks of daily operant access to ethanol and SACC, followed by an identical second deprivation period. P rats responded more for ethanol than SACC. All deprived groups increased responding on the ethanol lever, but not on the SACC lever. Daily consumption of 2 ml ethanol decreased the duration of the alcohol deprivation effect (ADE). Home cage access to 2 ml of SACC also decreased the ADE but to a lesser extent than access to ethanol. A second deprivation period further increased and prolonged the expression of an ADE. These results show ethanol is a more salient reinforcer than SACC. With concurrent access to ethanol and SACC, P rats do not display a saccharin deprivation effect. Depriving P rats of both ethanol and SACC had the most pronounced effect on the magnitude and duration of the ADE, suggesting that there may be some interactions between ethanol and SACC in their CNS reinforcing effects. PMID- 18400452 TI - Alcohol exposure on postnatal day 5 induces Purkinje cell loss and evidence of Purkinje cell degradation in lobule I of rat cerebellum. AB - The reduction in neuron number in specific brain regions is one of the most destructive aspects of alcohol-induced developmental brain injury, and its occurrence depends on the timing, pattern, and dose of maternal alcohol consumption during pregnancy. The purpose of this investigation was to quantify the dose-response aspect of Purkinje cell loss and rapid cellular degradation indicative of Purkinje cell loss following a single alcohol exposure on postnatal day 5 in lobule I, a lobule that has been shown to be vulnerable to alcohol induced injury during cerebellar development. Fluoro-Jade B was used to identify Purkinje cell degeneration in 2-h intervals during the first 24h following the single alcohol exposure. At the end of 24h, stereology cell counting techniques were used to estimate the number of Purkinje cells in lobule I of the cerebellum. Significant Fluoro-Jade B labeling of lobule I Purkinje cells began at 12-h postexposure in the 6.0-g/kg group with continued significant expression of the marker at the 16- and 18-h time points. Notably, the magnitude of Fluoro-Jade B expression in the 6.0-g/kg group remained high during the period between 12 and 24h even though the difference between the 6.0-g/kg group and other groups did not reach statistical significance at the 14-, 20-, and 24-h time points. On postnatal day 6, 24h following the alcohol exposure, rats exposed to the highest alcohol dose (6.0 g/kg) had lost significantly more Purkinje cells than those in the nutritional or caloric control to the highest dose of alcohol group. These results are suggestive of a unique relationship among the quantity of alcohol, the onset and duration of cell degradation, and the degree of eventual cell loss. Given that cerebellar Purkinje cells (and many developing neurons) are vulnerable to alcohol-induced neuronal loss within hours of a single alcohol insult, women should be counseled to avoid drinking alcohol in a manner that significantly increases blood alcohol levels during pregnancy (e.g., binge drinking). PMID- 18400453 TI - Alcohol reduces MMP-2 in humans and isolated smooth muscle cells. AB - Alcoholic beverages are known to exert a protective effect on atherosclerotic disease. This study aimed to assess the in vivo and in vitro effects of alcohol on matrix metalloproteinase (MMP) -2 and -9, known to determine atherosclerosis progression. Eighteen healthy volunteers, regular drinkers (two standard alcohol servings/day, on average) at first examination (baseline) were asked to abstain from any alcoholic beverage for one week (abstention), and then to assume two standard alcohol servings of beer daily for 1 week (re-exposure). Activity of MMP 2 and -9, total antioxidant activity (AOA), glutathione (GSH) plasma levels were carried out at baseline, at the end of abstention, and after 1 week of re exposure. To validate the in vivo results, MMP-2 activity and expression, AOA, and GSH, were determined in human smooth muscle cells treated for 96 h with increasing concentrations (12.5-100 mM) of ethanol. MMP-2, but not MMP-9 plasma activity was higher at abstention than at baseline or re-exposure (P<.001 and P< or =.005, respectively). Changes in AOA and GSH throughout the study were not significant. No correlation was found between MMPs and antioxidant activity. In vitro, ethanol at 25 mM reduced by around 10% MMP-2 activity (P=.003) in smooth muscle cells, whereas MMP-2 expression, AOA, and GSH were unaffected. Alcohol reduces MMP-2 plasma activity in healthy humans and in isolated vascular smooth muscle cells. This in vitro reduction is unrelated to MMP-2 expression in vascular cells or to antioxidant levels changes. PMID- 18400454 TI - Neuroprotective gene profile in the brain of magnesium-deficient mice. AB - BACKGROUND: Magnesium (Mg) deficiency may lead to serious metabolic, biological and organic dysfunctions, and cause various clinical disorders. In the current study, we explore endothelial cell activation, inflammation and cell death induced in the brain of adult mice by Mg-deficient diet. METHODS AND RESULTS: Neither TNFalpha, substance P, sTNFRI, sTNFRII proteins (ELISA), nor TNFalpha, adherence molecules and prolactin mRNAs, nor NK1R (immunohistochemistry on brain sections) were up-regulated. No inflammatory infiltrates and no apoptotic cells were observed. Using cDNA assay, we showed a neuroprotective, anti-apoptotic and neurotrophic gene expression profile in the brain at early stage of hypomagnesemia. As a model for neuronal injury, mild sound stimulation of Mg deficient mice without convulsive seizures triggers neither the release of substance P, nor the development of an inflammatory process or cell death in the brain. CONCLUSION: Our results suggest that Mg-deficiency in mice favours the development of a neuroprotective environment in the brain. PMID- 18400456 TI - Glutathione and Parkinson's disease: is this the elephant in the room? AB - At least 2 decades have past since the demonstration of a 40-50% deficit in total glutathione (GSH) levels in the substantia nigra in patients with Parkinson's disease (PD). The similar loss of GSH in the nigra in Incidental Lewy body disease, thought to be an early form of PD, indicates that this is one of the earliest derangements to occur in the pre-symptomatic stages of PD. Oxidative damage to lipids, protein and DNA in the nigra of PD patients is consistent with the loss of the antioxidant functions contributed by GSH. Past clinical trials that have used an antioxidant approach to treatment have used antioxidants that might substitute for GSH but these have shown modest to little benefit. More recent studies of the functions served by GSH in cells include in addition to its well-known participation in H(2)O(2) and toxin removal, such roles as modulation of protein function via thiolation which may control physiological and pathophysiological pathways to include DNA synthesis and repair, protein synthesis, amino acid transport, modulation of glutamate receptors and neurohormonal signaling. These multifunctional aspects to the workings of GSH in the cell would suggest that its loss perturbs many different processes and that replenishment and maintenance of GSH per se may be the best approach for preventing progressive damage from occurring. Despite this, few studies have been directed at specifically restoring GSH, although, as discussed herein, its unsanctioned use in PD is growing in popularity. This review will focus on glutathione in PD; the various functions carried out by glutathione and possible consequences of its depletion, as well as measures to elevate GSH in the CNS and its use in humans. Consideration of how the CNS generates and handles the substrates for GSH synthesis is also addressed with the view in mind that this may provide insights into control and maintenance of intracellular glutathione. PMID- 18400457 TI - [Heart failure with preserved ejection fraction]. AB - Heart failure with preserved ejection fraction (HFPEF) is defined by the combination of clinical symptoms or signs of heart failure and an ejection fraction> or =50+/-5%. HFPEF accounts for nearly 50% of cases of heart failure, and its prevalence is increasing as the population ages. It is seen most often as acute rapid-onset or flash pulmonary edema, in a patient with no or few symptoms except for these acute episodes, often associated with a precipitating factor. Diagnosis of HFPEF is facilitated by the development of plasma assays of natriuretic peptides and of Doppler echocardiography. Its causes, which must be identified, are mainly hypertensive, diabetic or ischemic cardiomyopathy, often combined. Its prognosis is similar to that of systolic heart failure and leads to frequent hospitalization for acute decompensation. Its treatment calls for the same therapeutic arsenal as for systolic heart failure, except among patients with diabetes. PMID- 18400455 TI - Inflammation and aging: can endocannabinoids help? AB - Aging often leads to cognitive decline due to neurodegenerative process in the brain. As people live longer, there exists a growing concern linked to long-term, slowly debilitating diseases, such as Alzheimer's disease for which a cure has not yet been found. Recently, the role of neuroinflammation has attracted attention due to its slow onset, chronic nature and its possible role in the development of many different neurodegenerative diseases. In the future, treatment of chronic neuroinflammation may help counteract aspects of neurodegenerative disease. Our recent studies have focused upon the endocannabinoid system for its unique effects on the expression of neuroinflammation. The basis for the manipulation of the endocannabinoid system in the brain in combination with existing treatments for Alzheimer's disease will be discussed in this review. PMID- 18400458 TI - Pain, distress, and adult-child interaction during venipuncture in pediatric oncology: an examination of three types of venous access. AB - This study examined pain and distress from needles in children undergoing blood sampling as a function of adult-child interaction and type of venous access (i.e., central external venous lines, internalized ports, or peripheral access via venipuncture). Participants were 55 pediatric oncology patients, aged 3-18 years, who were undergoing routine blood work. Pain ratings were obtained using the Faces Pain Scale-Revised (FPS-R) and conversation during the procedure was audio taped for coding using the Child-Adult Medical Procedure Interaction Scale Revised (CAMPIS-R). Children's ratings of pain using the FPS-R were similar in the port (M=2.57/10, standard deviation [SD]=3.46) and peripheral (M=2.56/10, SD=3.24) groups, despite the fact that most children with internal ports were given a topical anesthetic. Similarly, there were no differences between port and peripheral groups in rates of child coping or distress, or parent and nurse observations of child pain. As would be expected, external line access was not associated with pain or distress, even among very young children, suggesting that they appropriately understood the pain rating scale. Results of the transcribed CAMPIS-R data indicate that the influences in adult-child interaction are bidirectional. Support was found for the well-established positive relationship between child distress and adult reassurance and empathy. Implications for intervention and selection of central venous access devices are discussed. PMID- 18400459 TI - Assessing resident knowledge of acute pain management in hospitalized children: a pilot study. AB - This pilot study was undertaken to evaluate the hypotheses that there are differences in pediatric pain management (PPM) knowledge across resident specialties, that questions in the form of multiple-choice items could detect such differences, and that resident knowledge of analgesic-related adverse drug events (ADEs) would be greater than knowledge of PPM. Questions were based on two general categories of knowledge within acute pain management in hospitalized children: pediatric pain assessment and treatment, and identification of analgesic-related ADEs. As part of the pilot nature of this study, a convenience sample of 60 residents completed a 10-item PPM knowledge assessment prior to a PPM lecture. Twenty-six were pediatric residents (43%), 19 were orthopedic residents (32%), and 15 were anesthesiology residents (25%). All items had content validity. When controlling for resident year, performance by resident specialty was significantly different between anesthesia and orthopedics (P=0.006) and between anesthesia and pediatrics (P<0.001). Resident knowledge of analgesic-related ADEs was not greater than knowledge of PPM. The most difficult topics were opioid equianalgesia, assessment of the cognitively impaired child, and maximal acetaminophen doses. Repeated administration of the PPM knowledge assessment at multiple institutions will allow further evaluation of our initial findings, and with directed educational interventions, provide opportunity for measurement of improvement. PMID- 18400460 TI - Methodological challenges when using actigraphy in research. AB - Actigraphy has become a valuable clinical and research tool to objectively evaluate sleep, daytime activity, and circadian activity rhythms in healthy individuals as well as persons with primary and comorbid insomnia. However, procedures used for sampling, data processing, and analysis are not consistently reported in the literature. The wide variability in how actigraphy is reported makes it difficult to compare findings across studies. The procedures and reporting methods from 21 studies that used actigraphs to assess sleep and wake in adult patients with cancer are reviewed to highlight the differences in reporting strategies. Patients with cancer were chosen to illustrate the methodological challenges related to procedures and reporting in one population. The aim of this article was to advance standards of information presented in publications to enable comparisons across research studies that use actigraphy. Specific methodological challenges when using actigraphy in research include instrumentation, selection of pertinent variables, sampling, and data processing and analysis. Procedural decisions are outlined and discussed, and suggestions are made for standardized actigraphy information to include in research reports. More consistent procedures and reporting will advance the science of sleep, daytime activity, and circadian activity rhythms and their association with other health-related variables. PMID- 18400461 TI - A randomized controlled trial testing the efficacy of an HIV/AIDS symptom management manual. AB - This study investigates whether using an HIV/AIDS symptom management manual with self-care strategies for 21 common symptoms, compared to a basic nutrition manual, had an effect on reducing symptom frequency and intensity. A 775-person, repeated measures, randomized controlled trial was conducted over three months in 12 sites from the United States, Puerto Rico, and Africa to assess the relationship between symptom intensity with predictors for differences in initial symptom status and change over time. A mixed model growth analysis showed a significantly greater decline in symptom frequency and intensity for the group using the symptom management manual (intervention) compared to those using the nutrition manual (control) (t=2.36, P=0.018). The models identified three significant predictors for increased initial symptom intensities and in intensity change over time: (1) protease inhibitor-based therapy (increased mean intensity by 28%); (2) having comorbid illness (nearly twice the mean intensity); and (3) being Hispanic receiving care in the United States (increased the mean intensity by 2.5 times). In addition, the symptom manual showed a significantly higher helpfulness rating and was used more often compared to the nutrition manual. The reduction in symptom intensity scores provides evidence of the need for palliation of symptoms in individuals with HIV/AIDS, as well as symptoms and treatment side effects associated with other illnesses. The information from this study may help health care providers become more aware of self-management strategies that are useful to persons with HIV/AIDS and help them to assist patients in making informed choices. PMID- 18400462 TI - Characteristics and quality of life of patients who choose home care at the end of life. AB - Cancer patients with advanced disease and short-survival expectancy were given hospital-based advanced home care (AHC) or conventional care (CC), according to their preference. The two groups were compared at baseline to investigate whether there were differences between the AHC and the CC patients that may help explain their choice of care. The patients were consecutively recruited over 2(1/2) years. Sociodemographic and medical data, and the health-related quality of life (HRQL) of the two groups were compared. HRQL was assessed using a self-reporting questionnaire, including the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC QLQ-C30), the Impact of Event Scale (IES), five questions about social support, and two items concerning general well-being. The AHC group showed significantly poorer functioning on all the EORTC QLQ-C30 scales and an overall higher symptom burden than the CC patients. Fewer of the AHC patients were receiving cancer treatment. The AHC patients had lived longer with their cancer diagnosis, had a significantly shorter survival after study enrollment, and a significantly poorer performance status. The major differences between the two groups seemed to be related to being at different stages in their disease. The results indicate that patients are reluctant to accept home care until absolutely necessary due to severity of functioning impairments and symptom burden. These findings should be taken into consideration in planning palliative care services. PMID- 18400463 TI - High throughput kinetic Vibrio fischeri bioluminescence inhibition assay for study of toxic effects of nanoparticles. AB - Despite of the growing production and use of nanoparticles (NPs) in various applications, current regulations, including EC new chemical policy REACH, fail to address the environmental, health, and safety risks posed by NPs. This paper shows that kinetic Vibrio fischeri luminescence inhibition test--Flash Assay- that up to now was mainly used for toxicity analysis of solid and colored environmental samples (e.g. sediments, soil suspensions), is a powerful tool for screening the toxic properties of NPs. To demonstrate that Flash Assay (initially designed for a tube luminometer) can also be adapted to a microplate format for high throughput toxicity screening of NPs, altogether 11 chemicals were comparatively analyzed. The studied chemicals included bulk and nanosized CuO and ZnO, polyethylenimine (PEI) and polyamidoamine dendrimer generations 2 and 5 (PAMAM G2 and G5). The results showed that EC50 values of 30-min Flash Assay in tube and microplate formats were practically similar and correlated very well (log-logR2=0.98), classifying all analyzed chemicals, except nano CuO (that was more toxic in cuvette format), analogously when compared to the risk phrases of the EC Directive 93/67/EEC for ranking toxicity of chemicals for aquatic organisms. The 30-min EC50 values of nanoscale organic cationic polymers (PEI and dendrimers) ranged from 215 to 775 mg/l. Thirty-minute EC50 values of metal oxides varied largely, ranging from approximately 4 mg/l (bulk and nano ZnO) to approximately 100 mg/l (nano CuO) and approximately 4000 mg/l (bulk CuO). Thus, considering an excellent correlation between both formats, 96-well microplate Flash Assay can be successfully used for high throughput evaluation of harmful properties of chemicals (including organic and inorganic NPs) to bacteria. PMID- 18400464 TI - A bioluminescent cytotoxicity assay for assessment of membrane integrity using a proteolytic biomarker. AB - Measurement of cell membrane integrity has been widely used to assess chemical cytotoxity. Several assays are available for determining cell membrane integrity including differential labeling techniques using neutral red and trypan blue dyes or fluorescent compounds such as propidium iodide. Other common methods for assessing cytotoxicity are enzymatic "release" assays which measure the extra cellular activities of lactate dehydrogenase (LDH), adenylate kinase (AK), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in culture medium. However, all these assays suffer from several practical limitations, including multiple reagent additions, scalability, low sensitivity, poor linearity, or requisite washes and medium exchanges. We have developed a new cytotoxicity assay which measures the activity of released intracellular proteases as a result of cell membrane impairment. It allows for a homogenous, one-step addition assay with a luminescent readout. We have optimized and miniaturized this assay into a 1536 well format, and validated it by screening a library of known compounds from the National Toxicology Program (NTP) using HEK 293 and human renal mesangial cells by quantitative high-throughput screening (qHTS). Several known and novel membrane disrupters were identified from the library, which indicates that the assay is robust and suitable for large scale library screening. This cytotoxicity assay, combined with the qHTS platform, allowed us to quickly and efficiently evaluate compound toxicities related to cell membrane integrity. PMID- 18400466 TI - Effects of a third party observer and anxiety on tests of executive function. AB - For the past 10 years, research on the effects of observer presence on test performance has expanded in the neuropsychological literature. Previous studies have shown that the presence of a third party observer is associated with poorer performance on tests of effort, attention, concentration, learning, and memory. The present study was designed to investigate whether performance on tests of executive function is similarly impaired by the presence of a third party observer. The study also sought to examine associations among examinee anxiety, observer presence, and performance. Seventy-nine college undergraduates were recruited for the study, and 70 were included in the final analyses. Participants were randomly assigned to either the observation or control condition, and were administered verbal fluency tests, the Trail Making Test (parts A and B), and the Tactual Performance Test, as well as the Fear of Negative Evaluation scale and State-Trait Anxiety Inventory. Multivariate analyses of variance revealed that performance on the combined dependent variables was significantly associated with observer presence. A significant observation condition by trait anxiety interaction was also found. Univariate analyses revealed that performances on semantic fluency and TPT-localization were most strongly associated with observation and trait anxiety, with performance being poorer in the presence of a third party observer. Additionally, effects of trait anxiety on performance in the presence of an observer appear to vary depending on task characteristics. Implications and suggestions for further research are discussed. PMID- 18400465 TI - A bioassay for metals utilizing a human cell line. AB - The purpose of this study was to assess the ability of the HepG2 cell line to function as a bioassay for metal contamination in sediments, using metallothionein (MT) as a biomarker of exposure. Sediments were collected from the eastern and western ends of Lake Erie, extracted using EPA method 200.7, and analyzed for cadmium (Cd), mercury (Hg) and lead (Pb) levels using ICP-AES. Sediment extracts were neutralized then used at a 2.5% final concentration in the exposure medium. MT levels were measured using the cadmium-hemoglobin affinity assay after a 48 h exposure. Fortified blanks from the ICP protocol served as positive controls. Also, HepG2 cells were exposed to Cd, Pb or combinations of Cd and Pb to determine whether or not induction of MT observed in cells exposed to sediment extracts was due to a single metal, combinations of metals, pH, or some other factor. Additionally, cells were exposed to a range of Cd concentrations approximating the levels found in the extracts (0.0005-0.1mg/L) to determine if a concentration-response occurred. Total metal levels ranged from 527 to 33.5mg/kg with lead the predominant metal, accounting for 100-88.9% of the total quantifiable metals in the sediments. The biomarker response (MT induction) was strongly correlated (r2=0.9919, r2=0.990) with total metal and lead levels in the sediments, respectively, which supports recent field studies indicating the biomarker can discern differences in the strength of the inducing agent. Statistically significant MT induction was associated with sediments which contained measurable Cd concentrations and no significant differences were observed when comparing Cd only and Cd+Pb exposed cells indicating no interactions between Cd and Pb were occurring and supporting our finding that Cd was the main inducing agent in sediment extracts. MT levels also increased significantly in a concentration-dependent manner when cells were exposed only to Cd. Results suggest this human bioassay and the MT biomarker of exposure may be useful for monitoring complex metal mixtures in aquatic sediments. PMID- 18400467 TI - Different temperament and character dimensions correlate with panic disorder comorbidity in bipolar disorder and unipolar depression. AB - BACKGROUND: This study aimed to investigate temperament and character correlates of panic disorder (PD) comorbidity in euthymic patients with bipolar disorder (BD) or unipolar depression (UD). METHODS: Temperament and character were assessed using the Temperament and Character Inventory Revised (TCI-R) in 181 patients (70 patients with BD-I, 51 patients with BD-II and 60 with UD) in a euthymic state for at least 2 months. RESULTS: PD was diagnosed in 14.3% of BD-I patients, 31.4% of BD-II and 40% of UD. BD patients with PD, when compared with BD patients without PD, had higher scores on harm avoidance (OR=1.04; 95% CI=1.02 1.07; p=0.002). Patients with UD and PD, when compared to patients with UD without PD, had higher scores on social acceptance (OR=1.27; 95% CI=1.08-1.49; p=0.004). CONCLUSION: Different temperament and character dimensions correlated with PD comorbidity in BD and UD patients, suggesting different underlying pathophysiological mechanisms. PMID- 18400468 TI - Distribution and quantification of Candidatus Liberibacter americanus, agent of huanglongbing disease of citrus in Sao Paulo State, Brasil, in leaves of an affected sweet orange tree as determined by PCR. AB - Huanglongbing (HLB), an insect-transmitted disease of citrus, known for many years in Asia and Africa, has appeared in the state of Sao Paulo State (SSP), Brazil, in 2004, and the state of Florida, USA, in 2005. HLB endangers the very existence of citrus, as trees infected with the bacterial pathogen, irrevocably decline. In the absence of curative procedures, control of HLB is difficult and only based on prevention. Even though not available in culture, the HLB bacterium could be shown to be Gram-negative and to represent a new candidate genus, Candidatus Liberibacter, in the alpha subdivision of the Proteobacteria. Three Candidatus (Ca.) L. species occur: Ca. L. africanus in Africa, Ca. L. asiaticus in Asia, SSP, and Florida, and Ca. L. americanus in SSP. The liberibacters occur exclusively in the phloem sieve tubes. On affected trees, HLB symptoms are often seen on certain branches only, suggesting an uneven distribution of the Liberibacter. Occurrence of Ca. L. americanus, the major HLB agent in SSP, has been examined in 822 leaf samples from an affected sweet orange tree by two conventional PCR techniques and a newly developed real time (RTi) PCR, also used for quantification of the Liberibacter in the leaves. Even though RTi-PCR was able to detect as few as 10 liberibacters per gram of leaf tissue (l/g), no liberibacters could be detected in any of the many leaf samples from a symptomless branch, while in blotchy mottle leaves from symptomatic branches of the same tree, the Liberibacter titer reached values as high as 10(7)l/g. These results demonstrate the uneven distribution of the Liberibacter in HLB-affected trees. PMID- 18400469 TI - Guiding automated left ventricular chamber segmentation in cardiac imaging using the concept of conserved myocardial volume. AB - The active surface technique using gradient vector flow allows semi-automated segmentation of ventricular borders. The accuracy of the algorithm depends on the optimal selection of several key parameters. We investigated the use of conservation of myocardial volume for quantitative assessment of each of these parameters using synthetic and in vivo data. We predicted that for a given set of model parameters, strong conservation of volume would correlate with accurate segmentation. The metric was most useful when applied to the gradient vector field weighting and temporal step-size parameters, but less effective in guiding an optimal choice of the active surface tension and rigidity parameters. PMID- 18400470 TI - Nutrition disorder and immunologic parameters: study of the intestinal villi in growing rats. AB - OBJECTIVE: The aim of the present work was to study how a diet in which cereals were the only protein source would affect B and T lymphocytes and a cell population positive for thymus-expressed chemokine (TECK) in the intestinal villi of growing rats. METHODS: Wistar rats were fed a 6.5% precooked maize protein diet for 18-20 d (M group). An age-matched control group received stock diet (C group). Body weight (grams) was determined, ponderal growth rate (grams per day per 100 g) was calculated, and intestines were removed and processed by Saint Marie's technique. Tissue sections were studied by indirect immunofluorescence. CD5(+) T cells and the T-cell subsets TCRalphabeta(+), TCRgammadelta(+), CD4(+), CD8alpha(+), and CD8beta(+) in the lamina propria (LP) and intraepithelium, in addition to immunoglobulin A-positive B cells in the LP were determined (n cells/30 fields were read). In addition, the presence of the TECK(+) cell population was qualitatively assessed. RESULTS: The M versus C group showed statistically significant differences in body weight and ponderal growth rate. The number of immunoglobulin A-positive B cells in the LP and the CD5(+) T cells and CD4(+), CD8alpha(+), CD8beta(+), TCRalphabeta(+), and TCRgammadelta(+) T-cell subpopulations of the M group in the LP and intraepithelium of the gut villi were significantly decreased compared with the C group (P < 0.001). The M group also showed differences in the size and cellularity of the gut villi and in the distribution of TECK. CONCLUSION: The results show that intake of a low concentration of a low-quality dietary protein as the only source of protein produces an important disorder in the mucosal immunity of experimental rats. PMID- 18400471 TI - Expression of D-serine and glycine transporters in the prefrontal cortex and cerebellum in schizophrenia. AB - The NMDA receptor co-agonists D-serine and glycine are thought to contribute to glutamatergic dysfunction in schizophrenia. They are removed from the synapse by specific neuronal and glial transporters, the status of which is clearly relevant to theories of D-serine and glycine function in the disorder. D-serine is primarily transported by Asc-1, and glycine by GlyT1 but maybe also by SNAT2. As a first step to addressing this issue, we studied Asc-1, GlyT1 and SNAT2 expression in dorsolateral prefrontal cortex and cerebellum of 18 subjects with schizophrenia and 20 controls, using immunoblotting and in situ hybridization. Asc-1 protein and SNAT2 mRNA were decreased in schizophrenia in both regions. GlyT1 mRNA and protein, and Asc-1 mRNA, were not altered. Antipsychotic administration for 14 days did not alter expression of the genes in rat brain. Unchanged GlyT1 suggests that glycine transport is not markedly affected in schizophrenia, and therefore that increased synaptic removal is not the basis for the putative deficit in glycine modulation of NMDA receptors in the disorder. Lowered Asc-1 in schizophrenia implies that D-serine reuptake is reduced, perhaps as a response to decreased synaptic D-serine availability. However, this interpretation remains speculative. Further investigations will be valuable in the evaluation of these transporters as potential therapeutic targets in psychosis. PMID- 18400472 TI - Neuropsychological functioning in adolescents with first episode psychosis: a two year follow-up study. AB - Cognitive deficits are a core feature of psychotic disorders. Both in adult and adolescent populations, studies have shown that patients with psychosis have poorer cognitive functioning than controls. The cognitive domains that seem to be affected are mainly attention, working memory, learning and memory, and executive function. However, with regard to the trajectory of cognitive function throughout the illness, there is still a dearth of prospective data in patients who develop psychosis during adolescence. In this article, neuropsychological functioning was assessed in a sample of 24 first episodes of early onset psychosis (EOP) and 29 healthy adolescents at baseline and after a two-year follow-up. Patients with EOP showed lower scores than controls in overall cognitive functioning and in all specific domains assessed (attention, working memory, executive function, and learning and memory) both at baseline and the two-year follow-up. When changes in cognitive functioning over two years were assessed, patients and controls showed significant improvement in almost all cognitive domains. However, this improvement disappeared in the patient group after controlling for improvement in symptomatology. Our findings support a neurodevelopmental pathological process in this sample of adolescents with psychosis. PMID- 18400473 TI - Synthesis and characterization of PEO-PCL-PEO triblock copolymers: effects of the PCL chain length on the physical property of W(1)/O/W(2) multiple emulsions. AB - A series of poly(ethylene glycol)-block-poly(epsilon-caprolactone)-block poly(ethylene glycol) (PEO-PCL-PEO) triblock copolymers were prepared and then used for the investigation of the effects of the ratio of epsilon-caprolactone to poly(ethylene glycol) (i.e., [CL]/[EO]) on the physical properties of water-in oil-in-water (W(1)/O/W(2)) multiple emulsions containing a model reagent, ascorbic acid-2-glucoside (AA2G). In the synthesis, the [CL]/[EO] was varied from 0.11 to 0.31. The molecular weights and compositions of PEO-PCL-PEO were determined by GPC and (1)H NMR analyses. Thermal behavior and crystal formation were studied by DSC, XRD, FT-IR, and polarized optical microscopy (POM). Aggregate behavior of PEO-PCL-PEO was confirmed by DLS, UV, and (1)H NMR. Morphology and relative stiffness of the W(1)/O/W(2) multiple emulsions in the presence of PEO-PCL-PEO were studied by confocal laser scanning microscopy (CLSM) and rheometer. Variation in the [CL]/[EO] significantly affects the crystalline temperature and spherulite morphology of PEO-PCL-PEO. As the [CL]/[EO] increases, the CMCs of PEO-PCL-PEO decreases and the slope of aggregate size reduction against the copolymer concentration becomes steeper except for the lowest [CL]/[EO] value of PEO-PCL-PEO (i.e., P-222). P-222 significantly increases the viscosity of continuous (W(2)) phase, which implies the copolymer would exist in the W(2) phase. On the other hand, the triblock copolymers with relatively high [CL]/[EO] ratios mainly contribute to the size reduction of multiple emulsions and the formation of a firm wall structure. The particle size of the multiple emulsion decreases and the elastic modulus increased as [CL]/[EO] increases, confirmed by microscopic and rheometric analyses. PMID- 18400475 TI - Characterization of the grinding behaviour in a single particle impact device: studies on pharmaceutical powders. AB - The grinding behaviour of different materials can be described by the two material parameters fMat and Wm,min [Vogel, L., Peukert, W., 2003. Breakage behaviour of different materials-construction of a mastercurve for the breakage probability. Powder Technol. 129, 101-110]. fMat describes the resistance of particulate material against fracture in impact comminution, Wm,min characterizes the specific energy which a particle can take up without comminution. The material parameters are determined experimentally by single particle impact tests. This concept is also applicable to pharmaceutical powders, as will be shown in this work. A device is presented for the characterization of particles with sizes down to a few 10 microm. Particles are dispersed and accelerated in an air stream which is flowing against an impact plate. The impact velocity is controlled by the air flow. An LDA system enables the measurement of particle velocities. The results obtained with this jet mill are in accordance to those obtained from another single particle impact device used by Vogel and Peukert, in which the influence of fluid flow is completely avoided. Since the new device is especially designed for finer powders, it will allow a more detailed analysis of the material parameters at smaller particle sizes. Additionally, a new analysis method has been developed in order to determine the breakage probability not from sieve analysis but from laser light diffraction (LLD) data by using a population balance. PMID- 18400474 TI - Dilational viscoelasticity and relaxation properties of interfacial electrostatic complexes between oppositely charged hydrophobic and hydrophilic polyelectrolytes. AB - Strongly adsorbing hydrophobic cationic polyelectrolyte, Eudragit RS, containing approximately 2.5 mol% of pendent hydrophilic trimethylammonium (TMA) groups irreversibly adsorbs from its methylene chloride (MCl) solution at the MCl/water interface and forms solid-like adsorption layers (ALs). Submitted to periodic dilational deformations with the standard radial frequency omega(0)=0.63 rad/s, these ALs exhibit relatively high dilational storage modulus E' approximately 20 mN/m and practically zero loss modulus E'' at the bulk concentration C(Eud)=4 x 10(-3)g/L. The frequency scanning of these ALs in the diapason omega=0.01 0.63rad/s and the approximation of the experimental dependences E'(omega) and E''(omega) by two relaxation times rheological model makes it possible to estimate the crossing frequency of these ALs determined from the condition E'(omega(c))=E''(omega(c)) as omega(c) approximately 5 x 10(-4)rad/s. Upon dissolving the hydrophilic anionic polyelectrolyte, chitosan sulfate (ChS), in the water phase (C(ChS)=3 x 10(-2)g/L) the electrostatic interpolyelectrolyte complexes form at the MCl/water interface. The elasticity moduli E' and E'' of these mixed AL did not undergo remarkable variations, but the crossing frequency is sharply increased by approximately 10 times becoming equal to omega(c) congruent with 3 x 10(-3)rad/s. The increase of omega(c) certifies for the liquefaction of mixed Eudragit RS/ChS adsorption layers. A remarkable decrease of the storage modulus down to E'=8 mN/m and simultaneous increase of the crossing frequency up to omega(c) congruent with 10(-2)rad/s occurs upon increasing the concentrations of both components, Eudragit RS and ChS, up to 0.1g/L. The liquefaction effect in the mixed ALs of oppositely charged polyelectrolytes was explained on the basis of the proposed relaxation mechanism. The effect of the liquefaction of adsorption layers of strongly adsorbing hydrophobic polyelectrolytes by formation of interpolyelectrolyte complexes with hydrophilic polyelectrolytes must be taken into account in the production of nano-capsules and nano-fibers. PMID- 18400476 TI - [Ventricular repolarisation disorders and long QT syndrome: clinical and genetic study in 10 paediatric cases]. AB - We report 10 paediatric cases of ventricular repolarisation disorders and long QT syndrome, which differed in their mode of revelation, from asymptomatic forms to syncope events or heart arrest. Diagnosis is based on electrophysiological explorations and exhaustive genetic investigation. It allows a well-codified preventive and therapeutic action according to the genotype. PMID- 18400477 TI - [Dangers of early child-mother separations: Emile Zola as a precursor]. PMID- 18400478 TI - [Hypergonadotrophic hypogonadism and congenital galactosemia]. PMID- 18400480 TI - [Celiac disease diagnosis in 2008]. AB - Since the 1990's, the widespread use of very sensitive and specific serological tests has completely changed the conditions of the diagnosis of celiac disease (CD). The active form of the disease is now only the tip of the iceberg representing it. Currently, CD is evoked either in front of mild digestive symptoms at the usual age or in the course of screening in siblings of an index case or in patients at risk (insulin dependent diabetes for example) at a later age using IgA anti-endomysium or anti-tissue transglutaminase antibodies, the sensitivities and specificities of which exceed 90%. In some cases, HLA typing is helpful in allowing to exclude a patient who is neither DQ2 nor DQ8. The intestinal biopsy remains the "gold standard" of the diagnosis showing villous atrophy; the latter, however, is less and less often severe as the disease is milder and milder. The diagnosis of CD then rests on the confrontation of the clinical, biological and histological data. Once CD has been diagnosed, it can be classified either as active or silent (positive serology with isolated villous atrophy), or latent (positive serology with a normal mucosa) and the appropriate therapeutic decision can be taken. PMID- 18400479 TI - [Feeding during the first months of life and prevention of allergy]. AB - Allergy consists in the different manifestations resulting from immune reactions triggered by food or respiratory allergens. Both its frequency and severity are increasing. The easiest intervention process for allergy prevention is the reduction of the allergenic load which, for a major allergen such as peanuts, has to begin in utero. The primary prevention strategy relies first on the detection of at risk newborns, i.e. with allergic first degree relatives. In this targeted population, as well as for the general population, exclusive breastfeeding is recommended until the age of 6 months. The elimination from the mother's diet of major food allergens potentially transmitted via breast milk may be indicated on an individual basis, except for peanut, which is systematically retrieved. In the absence of breastfeeding, prevention consists in feeding at-risk newborns until the age of 6 months with a hypoallergenic formula, provided that its efficiency has been demonstrated by well-designed clinical trials. Soy based formulae are not recommended for allergy prevention. Complementary feeding should not be started before the age of 6 months. Introduction of egg and fish into the diet can be made after 6 months but the introduction of potent food allergens (kiwi, celery, crustaceans, seafood, nuts, especially tree nuts and peanuts) should be delayed after 1 year. This preventive policy seems partially efficacious on early manifestations of allergy but does not restrain the allergic march, especially in its respiratory manifestations. Probiotics, prebiotics as well as n-3 fatty polyunsaturated acids have not yet demonstrated any definitive protective effect. PMID- 18400482 TI - Morphological characteristics of sporangiospores of the tempe fungus Rhizopus oligosporus differentiate it from other taxa of the R. microsporus group. AB - The fungus Rhizopus oligosporus (R. microsporus var. oligosporus) is traditionally used to make tempe, a fermented food based on soybeans. Interest in the fungus has steadily increased, as it can also ferment other substrates, produce enzymes, and treat waste material. R. oligosporus belongs to the R. microsporus group consisting of morphologically similar taxa, which are associated with food fermentation, pathogenesis, or unwanted metabolite production (rhizonins and rhizoxins). The ornamentation pattern, shape, and size of sporangiospores of 26 R. microsporus group strains and two R. oryzae strains were studied using low-temperature SEM (LT-SEM) and LM. This study has shown that: (1) LT-SEM generates images from well-conserved sporangiophores, sporangia, and spores. (2) Robust spore ornamentation patterns can be linked to all different taxa of the R. microsporus group, some previously incorrectly characterized as smooth. Ornamentation included valleys and ridges running in parallel, granular plateaus, or smooth polar areas. Distribution of ornamentation patterns was related to spore shape, which either was regular, ranging from globose to ellipsoidal, or irregular. Specific differences in spore shape, size, and ornamentation were observed between Rhizopus taxa, and sometimes between strains. (3) R. oligosporus has a defect in the spore formation process, which may be related to the domesticated nature of this taxon. It had a high proportion, 10-31%, of large and irregular spores, and was significantly differentiated from other, natural Rhizopus taxa as evaluated with partial least squares discriminant analysis. It is remarkable that the vehicle of distribution, the sporangiospore, is affected in the strains that are distributed by human activity. This provides information about the specificity and speed of changes that occur in fungal strains because of their use in (food) industry. PMID- 18400481 TI - Choreography of Ig allelic exclusion. AB - Allelic exclusion guarantees that each B or T cell only produces a single antigen receptor, and in this way contributes to immune diversity. This process is actually initiated in the early embryo when the immune receptor loci become asynchronously replicating in a stochastic manner with one early and one late allele in each cell. This distinct differential replication timing feature then serves an instructive mark that directs a series of allele-specific epigenetic events in the immune system, including programmed histone modification, nuclear localization and DNA demethylation that ultimately bring about preferred rearrangement on a single allele, and this decision is temporally stabilized by feedback mechanisms that inhibit recombination on the second allele. In principle, these same molecular components are also used for controlling monoallelic expression at other genomic loci, such as those carrying interleukins and olfactory receptor genes that require the choice of one gene out of a large array. Thus, allelic exclusion appears to represent a general epigenetic phenomenon that is modeled on the same basis as X chromosome inactivation. PMID- 18400483 TI - Thiol redox state and related enzymes in sclerotium-forming filamentous phytopathogenic fungi. AB - Thiol redox state (TRS) reduced and oxidized components form profiles characteristic of each of the four main types of differentiation in the sclerotiogenic phytopathogenic fungi: loose, terminal, lateral-chained, and lateral-simple, represented by Rhizoctonia solani, Sclerotinia sclerotiorum, Sclerotium rolfsii, and Sclerotinia minor, respectively. A common feature of these fungi is that as their undifferentiated mycelium enters the differentiated state, it is accompanied by a decrease in the low oxidative stress-associated total reduced thiols and/or by an increase of the high oxidative stress associated total oxidized thiols either in the sclerotial mycelial substrate or in its corresponding sclerotium, indicating a relationship between TRS-related oxidative stress and sclerotial differentiation. Moreover, the four studied sclerotium types exhibit high activities of TRS-related antioxidant enzymes, indicating the existence of antioxidant protection of the hyphae of the sclerotium medulla until conditions become appropriate for sclerotium germination. PMID- 18400484 TI - "Slowed right down": Insights into the use of alcohol from research with Aboriginal Australians living with HIV. AB - OBJECTIVE: To describe the role that alcohol plays in the lives of Aboriginal people living with HIV in Western Australia. METHODS: Semi-structured interviews were conducted between February and September 2003 with 20 Aboriginal people who were HIV-positive; almost half the total number of Aboriginal people known to be living with HIV in Western Australia at that time. The main purpose of the study was to document their experiences of living with HIV and aspects of health service delivery. RESULTS: Drinking emerged as a key theme in the majority of interviews. Alcohol had a major role in disinhibition and risk-taking behaviour of both the participants and those they socialized with. It was perceived as a commodity, a way of altering reality, and a pathway through which social connection was maintained and was central to the common narratives of loss, chaos and transformation. Post-diagnosis, alcohol helped with disclosure and temporary blunting of distress as a result of the HIV diagnosis. It also contributed to a lack of capacity to comply with anti-retroviral therapy although narratives also included personal growth and restoration of physical health. CONCLUSIONS: Effective strategies to reduce risk of STIs and HIV and prevent other health problems need to address substance use issues but this also requires amelioration of the structural inequalities that make minority groups vulnerable. PMID- 18400485 TI - Effect of seasonal changes in quantities of biowaste on full scale anaerobic digester performance. AB - A 750,000l digester located in Roppen/Austria was studied over a 2-year period. The concentrations and amounts of CH4, H2, CO2 and H2S and several other process parameters like temperature, retention time, dry weight and input of substrate were registered continuously. On a weekly scale the pH and the concentrations of NH4+ -N and volatile fatty acids (acetic, butyric, iso-butyric, propionic, valeric and iso-valeric acid) were measured. The data show a similar pattern of seasonal gas production over 2 years of monitoring. The consumption of VFA and not the hydrogenotrophic CH4 production appeared to be the limiting factor for the investigated digestion process. Whereas the changes in pH and the concentrations of most VFA did not correspond with changes in biogas production, the ratio of acetic to propionic acid and the concentration of H2 appeared to be useful indicators for reactor performance. However, the most influential factors for the anaerobic digestion process were the amount and the quality of input material, which distinctly changed throughout the year. PMID- 18400486 TI - Effect of vadose zone on the steady-state leakage rates from landfill barrier systems. AB - Leakage rates are evaluated for a landfill barrier system having a compacted clay liner (CCL) underlain by a vadose zone of variable thickness. A numerical unsaturated flow model SEEP/W is used to simulate the moisture flow regime and steady-state leakage rates for the cases of unsaturated zones with different soil types and thicknesses. The results of the simulations demonstrate that harmonic mean hydraulic conductivity of coarse textured vadose zones is 3-4 orders of magnitude less than saturated hydraulic conductivity; whereas, the difference is only one order of magnitude for fine textured vadose zones. For both coarse and fine textured vadose zones, the effective hydraulic conductivity of the barrier system and the leakage rate to an underlying aquifer increases with increasing thickness of the vadose zone and ultimately reaches an asymptotic value for a coarse textured vadose zone thickness of about 10m and a fine textured vadose zone thickness of about 5m. Therefore, the fine and coarse textured vadose zones thicker than about 5m and 10m, respectively, act as an effective part of the barrier systems examined. Although the thickness of vadose zone affects the effective hydraulic conductivity of the overall barrier system, the results demonstrated that the hydraulic conductivity of the CCL is the dominant factor controlling the steady-state leakage rates through barrier systems having single low permeability clay layers. PMID- 18400487 TI - The complexities of starch biosynthesis in cereal endosperms. AB - Starch serves not only as an energy source for plants, animals, and humans but also as an environmentally friendly alternative for fossil fuels. Here, we describe recent findings concerning the synthesis of this important molecule in the cereal endosperm. Results from six separate transgenic reports point to the importance of adenosine diphosphate glucose pyrophosphorylase in controlling the amount of starch synthesized. The unexpected cause underlying the contrast in sequence divergence of its two subunits is also described. A major unresolved question concerning the synthesis of starch is the origin of nonrandom or clustered alpha-1,6 branch-points within the major component of starch, amylopectin. Developing evidence that several of the starch biosynthetic enzymes involved in amylopectin synthesis occur in complexes is reviewed. These complexes may provide the specificity for the formation of nonrandom branch-points. PMID- 18400488 TI - Introduction of additional double reading of mammograms by radiographers: effects on a biennial screening programme outcome. AB - PURPOSE: To determine the effect of introducing radiographer double reading, in addition to standard radiologist double reading, on screening mammography outcome. METHODS: In period A, 66,225 mammograms were read by two screening radiologists. In period B, 78,325 mammograms were read by two radiographers in addition and radiologists were blinded to the referral opinion of the radiographers. Mammograms, for which only radiographers had suggested referral, (i.e. cases that would only be referred by technologists) were re-evaluated by the screening radiologists. Women were referred if at least one radiologist considered this necessary, and diagnostic costs of these additional referrals were estimated. RESULTS: In period A, 322 cancers were diagnosed after referral of 678 women. During period B, radiologists initially referred 1122 patients and 411 cancers were detected. Radiologists' referral rate was higher in period B than in period A (1.43% versus 1.02%, p<0.001), as well as the cancer detection rate per 1000 women screened (CDR) (5.25 versus 4.86, p=0.3). The positive predictive value of referral (PPV) was 36.6% versus 47.5% (p<0.001). In period B, radiologist review of 544 additional positive radiographer readings led to 102 extra referrals, with 29 additional cancers detected, resulting in an overall referral rate of 1.56% (compared to period A, p<0.001), an overall CDR of 5.62 (p=0.048) and an overall PPV of 35.9% (p<0.001). Workup expenses of the 102 additional referrals were euro60,274. CONCLUSION: Additional radiographer double reading detected cancers that would have been missed by radiologists. Mean expenses for diagnostic confirmation of these extra cancers was euro2078 per cancer. PMID- 18400489 TI - Hormonal regulation of spermatogenesis and spermiogenesis. AB - Normal testicular function is dependent upon hormones acting through endocrine and paracrine pathways both in vivo and in vitro. Sertoli cells provide factors necessary for the successful progression of spermatogonia into spermatozoa. Sertoli cells have receptors for follicle stimulating hormone (FSH) and testosterone which are the main hormonal regulators of spermatogenesis. Hormones such as testosterone, FSH and luteinizing hormone (LH) are known to influence the germ cell fate. Their removal induces germ cell apoptosis. Proteins of the Bcl-2 family provide one signaling pathway which appears to be essential for male germ cell homeostasis. In addition to paracrine signals, germ cells also depend upon signals derived from Sertoli by direct membrane contact. Somatostatin is a regulatory peptide playing a role in the regulation of the proliferation of the male gametes. Activin A, follistatin and FSH play a role in germ cell maturation during the period when gonocytes resume mitosis to form the spermatogonial stem cells and differentiating germ cell populations. In vitro cultures systems have provided evidence that spermatogonia in advance stage of differentiation have specific regulatory mechanisms that control their fate. This review article provides an overview of the literature concerning the hormonal pathways regulating spermatogenesis. PMID- 18400490 TI - Aldosterone and the vascular system. AB - Aldosterone can act in different tissues exerting physiological and pathological effects. At the vascular level, aldosterone affects endothelial function since administration of aldosterone impaired endothelium-dependent relaxations. In addition, the administration of mineralocorticoid receptor antagonists ameliorate relaxation to acetylcholine in models of both hypertension and atherosclerosis and in patients with heart failure. A reduction in nitric oxide levels seems to be the main mechanism underlying this effect due to a reduction in its production as well as an increase in its degradation by reactive oxygen species. Aldosterone is a pro-inflammatory factor that can participate in the vascular inflammatory process associated with different pathologies including hypertension through activation of the NFkappaB system, which mediates the vascular production of different cytokines. This mineralocorticoid also participates in the vascular remodeling observed in hypertensive rats since the administration of eplerenone improved the media-to-lumen ratio in these animals. This effect seems to be due to an increase in extracellular matrix. In summary, aldosterone through mineralocorticoid receptors can participate in the vascular damage associated with different pathologies including hypertension through its prooxidant, pro inflammatory and profibrotic effects that triggered endothelial dysfunction, an inflammatory process and vascular remodeling. PMID- 18400492 TI - Fate of 14C-Pyrene in soil-plant system amended with pig manure compost and Tween 80: a growth chamber study. AB - This paper evaluated the effects of a pig manure compost (PMC) and a nonionic surfactant Tween 80 on the fate of 14C-Pyrene (Pyr) in a soil-plant system (Agropyron elongatum). Soils spiked with 14C-4, 5, 9, 10-Pyr were amended with 7.5% (w/w) PMC together with or without 100mgkg(-1) of Tween 80. Unplanted soil without amendments was set as the control. Gas phases of the systems were monitored for 14CO2 over a 60 days period. The impact of PMC and Tween 80 on the apparent loss of the PAH and the distribution of 14C-activity in the systems was studied. 14C-activity associated with different soil fractions was further examined by using methyl-isobutyl-ketone (MIBK) fractionation method. The results showed that the addition of PMC could increase the dissipation of Pyr in vegetated soil from 12.1% to 58.7%, while the co-addition of Tween 80 and PMC could further enhance the dissipation to 90.3%. Pyr dissipation in soil was correlated with the mineralization of 14C-Pyr, indicating that Pyr dissipation was mainly due to mineralization. A higher formation of water-extractable metabolites was observed in soil amended with PMC and Tween 80, and this was correlated with a higher biomass accumulation of 14C-activity and higher bound residue formation in the soil. Overall, this study suggested that the co application of PMC and Tween 80 could improve phytoremediation of Pyr contaminated soil. PMID- 18400491 TI - Ligand structure-dependent activation of estrogen receptor alpha/Sp by estrogens and xenoestrogens. AB - This study investigated the effects of E2, diethylstilbestrol (DES), antiestrogens, the phytoestrogen resveratrol, and the xenoestrogens octylphenol (OP), nonylphenol (NP), endosulfan, kepone, 2,3,4,5-tetrachlorobiphenyl-4-ol (HO PCB-Cl(4)), bisphenol-A (BPA), and 2,2-bis-(p-hydroxyphenyl)-1,1,1 trichloroethane (HPTE) on induction of luciferase activity in breast cancer cells transfected with a construct (pSp1(3)) containing three tandem GC-rich Sp binding sites linked to luciferase and wild-type or variant ERalpha. The results showed that induction of luciferase activity was highly structure-dependent in both MCF 7 and MDA-MB-231 cells. Moreover, RNA interference assays using small inhibitory RNAs for Sp1, Sp3 and Sp4 also demonstrated structure-dependent differences in activation of ERalpha/Sp1, ERalpha/Sp3 and ERalpha/Sp4. These results demonstrate for the first time that various structural classes of ER ligands differentially activate wild-type and variant ERalpha/Sp-dependent transactivation, selectively use different Sp proteins, and exhibit selective ER modulator (SERM)-like activity. PMID- 18400493 TI - PVA-gel beads enhance granule formation in a UASB reactor. AB - PVA-gel beads were used as a biocarrier in a lab-scale UASB reactor treating synthetic wastewater composed of corn steep liquor (CSL) with the aim of evaluating its use as a growth nucleus to enhance granule formation. Over 117 days of operation, the organic loading rate was increased to 22.5kgCOD/m3/day with an influent COD of about 10.8g/L at an HRT of 12h with COD removal efficiencies greater than 87%. By the end of the study period, the PVA-gel turned black and granule formation was achieved as compared with the formation of much fewer natural granules without the PVA-gel nucleus. No filamentous bacteria were found on the surface or interior of the PVA-gel beads. The PVA-gel granules had an average settling velocity 200m/h (5cm/s), and a biomass attachment of 0.93g VSS/g PVA-gel. The required time for formation of PVA-gel granules was thus demonstrated to be shorter than that of ordinary sludge granules under the experimental conditions used in this study. PMID- 18400494 TI - Pile settlement and volume reduction measurement during forced-aeration static composting. AB - In this study, a settlement measurement device was used to track the settlement of different layers in composting piles, and time courses of settlement velocity, bulk density distribution, and effective bulk density were investigated. Settlement is divided into two stages: the physical compressive settlement and the mass loss settlement. At the end of the former stage, bulk density (rho') and pile height (h) follow rho'=Ah(B), where A is a fitting parameter for the initial bulk density of the composting material, and B is the compression parameter. The physical compressibilities of composting mixtures are night-soil sludge and garden refuse (1:1) >sewage sludge and cornstalks (5:4) >night-soil sludge and garden refuse (2:3) >sewage sludge and bark (1:1). Adding the proportion of bulking agent in the proper extent effectively retarded composting settlement. Bark exhibits a better supporting capability than cornstalk does as a bulking agent. PMID- 18400495 TI - Fragment-based discovery of hepatitis C virus NS5b RNA polymerase inhibitors. AB - Non-nucleoside inhibitors of HCV NS5b RNA polymerase were discovered by a fragment-based lead discovery approach, beginning with crystallographic fragment screening. The NS5b binding affinity and biochemical activity of fragment hits and inhibitors was determined by surface plasmon resonance (Biacore) and an enzyme inhibition assay, respectively. Crystallographic fragment screening hits with approximately 1-10mM binding affinity (K(D)) were iteratively optimized to give leads with approximately 200nM biochemical activity and low microM cellular activity in a Replicon assay. PMID- 18400496 TI - Novel 2-imidazoles as potent and selective alpha1A adrenoceptor partial agonists. AB - Novel 2-imidazoles have been identified as potent partial agonists of the alpha(1A) adrenergic receptor, with good selectivity over the alpha(1B), alpha(1D) and alpha(2A) receptor sub-types. Sulfonamide 23 possessed attractive drug-like properties with respect to physicochemical and ADME properties and wide ligand selectivity. PMID- 18400497 TI - Discovery of amino-acetonitrile derivatives, a new class of synthetic anthelmintic compounds. AB - A new series of amino-acetonitrile derivatives (AAD) have been discovered that exhibit high anthelmintic activity against parasitic nematode species such as Haemonchus contortus and Trichostrongylus colubriformis. Significantly, these compounds also demonstrate activity against nematode strains resistant to the currently available broad-spectrum anthelmintics. The discovery, synthesis, structure-activity relationship and biological results are presented. PMID- 18400498 TI - Design, synthesis and evaluation of 2-phenyl-1H-benzo[d]imidazole-4,7-diones as vascular smooth muscle cell proliferation inhibitors. AB - A series of 2-phenyl-1H-benzo[d]imidazole-4,7-diones were synthesized and tested for their inhibitory activity on the PDGF-stimulated proliferation of rat aortic vascular smooth muscle cells. Among the tested compounds, 6-arylthio-5-chloro-2 phenyl-1H-benzo[d]imidazole-4,7-diones exhibited an potent antiproliferative activity. PMID- 18400499 TI - Selective androgen receptor modulators based on a series of 7H-[1,4]oxazino[3,2 g]quinolin-7-ones with improved in vivo activity. AB - Modification on a lead series of [1,4]oxazino[3,2-g]quinolin-7-ones at the 2 position led to selective androgen receptor modulators with improved in vivo activity. The most potent analog (-)-33a exhibited full maintenance of levator ani muscle at 3mg/kg and reduced activity on ventral prostate weight in a 2-week orally-dosed and orchidectomized rat maintenance assay. PMID- 18400500 TI - The effects of age on medio-lateral stability during normal and narrow base walking. AB - We examined age-related differences in frontal plane stability during performance of narrow base (NB) walking relative to usual gait. A cross-sectional analysis of participants from the Baltimore Longitudinal Study of Aging (BLSA) was performed on data from the BLSA Motion Analysis Laboratory. Participants were 34 adults aged 54-92 without history of falls. We measured step error rates during NB gait and spatial-temporal parameters, frontal plane stability, and gait variability during usual and NB gait. There was a non-significant age-associated linear increase in step error rate (P=0.12) during NB gait. With increasing age, step width increased (P=0.002) and step length and stride velocity decreased (P<0.001), especially during NB gait. Age-associated increases in medio-lateral (M-L) center of mass (COM) peak velocity (P<0.001) and displacement (P=0.005) were also greater during NB compared to usual gait. With increasing age there was greater variability in stride velocity (P=0.001) and step length (P<0.001) under both conditions. Age-associated differences related to M-L COM stability suggest that the quantification of COM control during NB gait may improve identification of older persons at increased falls risk. PMID- 18400501 TI - Assessment of skin extensibility and joint hypermobility in patients with spontaneous cervical artery dissection and Ehlers-Danlos syndrome. AB - Ehlers-Danlos syndrome (EDS) is a hereditary connective tissue disorder. One important clinical characteristic of classical type EDS is skin hyperextensibility. Examination of clinical evidence and electron microscopic views of skin biopsies suggest that connective tissue abnormalities resembling very mild EDS are present in a sizable proportion of patients with spontaneous cervical artery dissection (sCAD). Manual assessment of skin extensibility is difficult. Therefore, non-invasive machine-aided measurement of skin extensibility was used and compared with manual assessment of skin extensibility and joint hyperextensibility. Patients with classical EDS, vascular-type EDS, sCAD and healthy patients were evaluated. Skin extensibility was measurably and palpably elevated in all patients with classical type EDS but not in sCAD patients or patients with vascular-type EDS compared to healthy control individuals. Our method is able to measure the increased skin extensibility in classical type EDS. Increased skin extensibility is not present in sCAD patients. PMID- 18400502 TI - Synthesis and activity of tetrapeptidic HTLV-I protease inhibitors possessing different P3-cap moieties. AB - The causative agent behind adult T-cell leukemia and tropical spastic paraparesis/HTLV-I-associated myelopathy is the human T-cell leukemia virus type 1 (HTLV-I). Tetrapeptidic HTLV-I protease inhibitors were designed on a previously reported potent inhibitor KNI-10516, with modifications at the P(3) cap moieties. All the inhibitors showed high HIV-1 protease inhibitory activity (over 98% inhibition at 50nM) and most exhibited highly potent inhibition against HTLV-I protease (IC(50) values were less than 100nM). PMID- 18400503 TI - A quantitative study of sodium tungstate protective effect on pancreatic beta cells in streptozotocin-induced diabetic rats. AB - Diabetes is a major public health problem. Development of new therapies that are able to improve glycemia management, cure diabetes, and can even protect from it, are of great interest. This study investigated the protective effect of sodium tungstate against STZ-induced beta-cell damages by means of stereological methods. Sixty rats were divided into six groups: control (C), tungstate-treated control (TC), STZ-induced diabetic (D), STZ-induced diabetic rats were treated by sodium tungstate from 1 week before STZ injection (TDB), food-restricted diabetic (FRD), and diabetic rats treated with sodium tungstate 1 week after STZ administration (TDA). Stereological estimation of pancreas volume, islets volume density, volume-weighted mean islets volume and mass of beta cells, islets, and pancreas and total number of islets were done. Islets volume density, volume weighted mean islets volume, and mass of beta cells, islets, and pancreas of TDB group was significantly higher than D, FRD and TDA groups (P<0.001) and was comparable to controls (C and TC groups). Total number of islets, pancreas wet weight and volume did not show any significant changes between these groups (P>0.05). Results suggested that sodium tungstate preserves pancreatic beta cells from STZ-induced damages and diabetes induction in rats. PMID- 18400504 TI - In-situ SEM indentation studies of the deformation mechanisms in TiN, CrN and TiN/CrN. AB - In this study, the microstructure and the deformation mechanisms of TiN, CrN and multilayer TiN/CrN thin films on silicon substrates were investigated. Cross sectional lamellas of nanoindents were prepared by focused ion beam milling to observe by transmission electron microscopy the microstructure of the as deposited and deformed materials. TiN film exhibits nanocrystalline columns, whereas CrN shows large grains. The TiN/CrN multilayer presents microstructural features typical for both materials. A film hardness of 16.9GPa for CrN, 15.8GPa for TiN and 16.6GPa for TiN/CrN was found by the nanoindentation. Reduced modulus recorded for TiN and CrN reference coatings were 221.54 and 171.1GPa, respectively, and 218.6GPa for the multilayer coating. The deformation mechanisms were observed via in-situ scanning electron microscope nanoindentation. The TiN thin film showed short radial cracks, whereas CrN deformed through pile-up and densification of the material. For TiN/CrN multilayer pile-up and cracks were found. Transmission electron microscopy observations indicated that TiN deforms through grain boundary sliding and CrN via densification and material flow. The deformation mechanism observed in TiN/CrN multilayer was found to be a mixture of both modes. PMID- 18400505 TI - Differential expression of myogenic regulatory factor MyoD in pacu skeletal muscle (Piaractus mesopotamicus Holmberg 1887: Serrasalminae, Characidae, Teleostei) during juvenile and adult growth phases. AB - Skeletal muscle is the edible part of the fish. It grows by hypertrophy and hyperplasia, events regulated by differential expression of myogenic regulatory factors (MRFs). The study of muscle growth mechanisms in fish is very important in fish farming development. Pacu (Piaractus mesopotamicus) is one of the most important food species farmed in Brazil and has been extensively used in Brazilian aquaculture programs. The aim of this study was to analyze hyperplasia and hypertrophy and the MRF MyoD expression pattern in skeletal muscle of pacu (P. mesopotamicus) during juvenile and adult growth stages. Juvenile (n=5) and adult (n=5) fish were anaesthetized, sacrificed, and weight (g) and total length (cm) determined. White dorsal region muscle samples were collected and immersed in liquid nitrogen. Transverse sections (10 microm thick) were stained with Haematoxilin-Eosin (HE) for morphological and morphometric analysis. Smallest fiber diameter from 100 muscle fibers per animal was calculated in each growth phase. These fibers were grouped into three classes (<20, 20-50, and >50 microm) to evaluate hypertrophy and hyperplasia in white skeletal muscle. MyoD gene expression was determined by semi-quantitative RT-PCR. PCR products were cloned and sequenced. Juvenile and adult pacu skeletal muscle had similar morphology. The large number of <20 microm diameter muscle fibers observed in juvenile fish confirms active hyperplasia. In adult fish, most fibers were over 50 microm diameter and denote more intense muscle fiber hypertrophy. The MyoD mRNA level in juveniles was higher than in adults. A consensus partial sequence for MyoD gene (338 base pairs) was obtained. The Pacu MyoD nucleotide sequence displayed high similarity among several vertebrates, including teleosts. The differential MyoD gene expression observed in pacu white muscle is possibly related to differences in growth patterns during the phases analyzed, with hyperplasia predominant in juveniles and hypertrophy in adult fish. These results should provide a foundation for understanding the molecular control of skeletal muscle growth in economically important Brazilian species, with a view to improving production quality. PMID- 18400506 TI - High resolution imaging of craniofacial bone specimens by flat-panel volumetric computed tomography. AB - INTRODUCTION: The prototype flat-panel volumetric computed tomograph (fpvCT) provides a new 3D imaging technology with detailed high resolution by using large area flat-panel X-ray detectors. The object of this study was to evaluate the benefit of high resolution imaging using the experimental fpvCT to visualise different types of human craniofacial bone pathology. The study proved the feasibility of performing an intraoperative evaluation of free margins in bone malignancies using fpvCT. MATERIAL AND METHODS: In this study, 35 bone specimens of various pathological types were examined by fpvCT. fpvCT data were compared with pre-operative multislice clinical CT images as well as with post-operative histological findings. RESULTS: Bone tumours can be visualised with their specific pathological architecture and infiltration structure faster and more precisely by fpvCT than by multislice CT. The analysis of the resection margins supports the surgical procedure intraoperatively, especially when an immediate reconstruction with bone transplantation is carried out. DISCUSSION: The fpvCT has a superior image quality when compared with clinical CT systems. The imaging of the bone structure itself has been shown to be useful for the interpretation of osseous resection borders. Furthermore, it can facilitate the diagnosis of tumour progression, especially in areas that are difficult to access, such as the base of the skull. PMID- 18400507 TI - Cancer vaccines: accomplishments and challenges. AB - Advancements in knowledge in diverse fields of science, including genetics, cell biology, molecular biology and biochemistry, have shed light on the origins of cancer and cell intrinsic properties that allow it to grow, invade and metastasize. Many therapies currently in use or under development are based on this knowledge. Advances in immunology, on the other hand, have shed light on how the host responds to these malignant properties of cancer. Based on that knowledge, immunotherapy, in particular vaccines directed at improving the host response against cancer, is being developed as an alternative therapeutic approach. In this review, we address main issues that have driven development of cancer vaccines and the challenges that have been met and/or are anticipated. PMID- 18400508 TI - Cetuximab efficacy and safety in a retrospective cohort of elderly patients with heavily pretreated metastatic colorectal cancer. AB - BACKGROUND: Few data are available from clinical trials for elderly patients receiving cetuximab. PATIENTS AND METHODS: The clinical data of consecutive patients aged > or =70 years given cetuximab for metastatic CRC were retrospectively captured from hospital pharmacy registries in seven centers. RESULTS: Fifty-six patients received cetuximab+/-with irinotecan. Median age was 76 years (70-84), 86% of patients were pretreated with fluoropyrimidines, irinotecan and oxaliplatin and 69.6% had documented resistance to irinotecan. Objective response rate was 21% (95% CI: 11-32%). The median progression-free survival was 4.4 months (95% CI: 3.0-5.7 months) and the median overall survival was 16.0 months (95% CI: 13.5-18.5 months). Skin rash occurred in 75% of the patients (11% grade 3) and diarrhea in 80% (20% grades 3-4). CONCLUSION: Tolerability of cetuximab was acceptable in elderly patients with pretreated metastatic CRC. Efficacy appeared similar to that observed in younger patients. PMID- 18400510 TI - Effect of anti-tumor necrosis factor-alpha therapies on body mass index in patients with psoriasis. AB - Psoriasis is a lifelong, chronic and immune-mediated skin disease affecting approximately 1-3% of the Caucasian population. Pathogenesis of psoriasis is associated with an increased expression of tumor necrosis factor-alpha (TNF alpha). TNF-alpha is a pro-inflammatory cytokine and important mediator of cachexia. Anti-TNF-alpha therapies are effective in the treatment of psoriasis. The primary end-point was to investigate retrospectively the effect of anti-TNF therapies on body weight and body mass index (BMI) in patients with psoriasis under treatment with infliximab, etanercept, adalimumab (anti-TNF-alpha group), efalizumab or methotrexate (control group). The patients were treated for 48 weeks. BMI, weight and disease activity were measured at baseline (week 0), weeks 12, 24 and 48. At week 24 a significant increase in body weight and BMI in the anti-TNF-alpha treatment group compared to the control was observed. BMI and body weight did not interfere with the drugs' efficacy. We report a significant weight gain associated with three different anti-TNF-alpha therapies in a large number of patients affected with psoriasis. PMID- 18400509 TI - Granulocyte colony-stimulating factor: molecular mechanisms of action during steady state and 'emergency' hematopoiesis. AB - Neutrophils are phagocytes whose principal function is to maintain anti-bacterial immunity. Neutrophils ingest and kill invading bacteria, releasing cytotoxic, chemotactic and inflammatory mediators at sites of infection. This serves to control the immediate host immune response and attract other cells, such as macrophages and dendritic cells, which are important for establishing long-term adaptive immunity. Neutrophils thus contribute to both the initiation and the maintenance of inflammation at sites of infection. Aberrant neutrophil activity is deleterious; suppressed responses can cause extreme susceptibility to infection while overactivation can lead to excessive inflammation and tissue damage. This review will focus on neutrophil regulation by granulocyte colony stimulating factor (G-CSF), the principal cytokine controlling neutrophil development and function. The review will emphasize the molecular aspects of G CSF-driven granulopoiesis in steady state (healthy) conditions and during demand driven or 'emergency' conditions elicited by infection or clinical administration of G-CSF. Understanding the molecular control of granulopoiesis will aid in the development of new approaches designed to treat disorders of neutrophil production and function. PMID- 18400511 TI - Time-resolved limited proteolysis of mitogen-activated protein kinase-activated protein kinase-2 determined by LC/MS only. AB - Mass spectrometry has gained prominence in limited proteolysis studies largely due to its unparalleled precision in determining protein molecular mass. However, proteolytic fragments usually cannot be identified through direct mass measurement, since multiple subsequences of a protein can frequently be matched to observed masses of proteolytic fragments. Therefore, additional information from N-terminal sequencing is often needed. Here we demonstrate that mass spectrometry analysis of the time course of limited proteolysis reactions provides new information that is self-sufficient to identify all proteolytic fragments. The method uses a non-specific protease like subtilisin and exploits information contained in the time-resolved dataset such as: increased likelihood of identifying larger fragments generated during initial proteolysis solely by their masses, additivity of the masses of two mutually exclusive sequence regions that generate the full-length molecule (or an already assigned subfragment), and analyses of the proteolytic subfragment patterns that are facilitated by having established the initial cleavage sites. We show that the identities of the observed proteolytic fragments can be determined by LC/MS alone because enough constraints exist in the time-resolved dataset. For a medium-sized protein, it takes about 8 h to complete the study, a significant improvement over the traditional SDS-PAGE and N-terminal sequencing method, which usually takes several days. We illustrate this method with application to the catalytic domain of mitogen-activated protein kinase-activated protein kinase-2, and compare the results with N-terminal sequencing data and the known X-ray crystal structure. PMID- 18400512 TI - Probing the gas-phase folding kinetics of peptide ions by IR activated DR-ECD. AB - The effect of infrared (IR) irradiation on the electron capture dissociation (ECD) fragmentation pattern of peptide ions was investigated. IR heating increases the internal energy of the precursor ion, which often amplifies secondary fragmentation, resulting in the formation of w-type ions as well as other secondary fragments. Improved sequence coverage was observed with IR irradiation before ECD, likely due to the increased conformational heterogeneity upon IR heating, rather than faster breakdown of the initially formed product ion complex, as IR heating after ECD did not have similar effect. Although the ECD fragment ion yield of peptide ions does not typically increase with IR heating, in double resonance (DR) ECD experiments, fragment ion yield may be reduced by fast resonant ejection of the charge reduced molecular species, and becomes dependent on the folding state of the precursor ion. In this work, the fragment ion yield was monitored as a function of the delay between IR irradiation and the DR-ECD event to study the gas-phase folding kinetics of the peptide ions. Furthermore, the degree of intracomplex hydrogen transfer of the ECD fragment ion pair was used to probe the folding state of the precursor ion. Both methods gave similar refolding time constants of approximately 1.5 s(-1), revealing that gaseous peptide ions often refold in less than a second, much faster than their protein counterparts. It was also found from the IR-DR-ECD study that the intramolecular H. transfer rate can be an order of magnitude higher than that of the separation of the long-lived c/z product ion complexes, explaining the common observation of c. and z type ions in ECD experiments. PMID- 18400513 TI - Expression of a bioactive fusion protein of Escherichia coli heat-labile toxin B subunit to a synapsin peptide. AB - The B subunit of Escherichia coli heat-labile toxin (LTB) may function as an efficient carrier molecule for the delivery of genetically coupled antigens across the mucosal barrier. We constructed vectors for the expression of LTB and LTBSC proteins. LTBSC is a fusion protein that comprises the amino acid sequence from the C-domain of rat synapsin fused to the C-terminal end of LTB. Both constructions have a coding sequence for a 6His-tag fused in-frame. LTBSC was expressed in E. coli as inclusion bodies. The inclusion bodies were isolated and purified by Ni2+-chelating affinity chromatography under denaturing condition. Purified LTBSC was diluted in several refolding buffers to gain a soluble and biologically active protein. Refolded LTBSC assembled as an active oligomer which binds to the GM1 receptor in an enzyme-linked immunosorbent assay (ELISA). Soluble LTB in the E. coli lysate was also purified by Ni2+-chelating affinity chromatography and the assembled pentamer was able to bind with high affinity to GM1 in vitro. LTBSC and LTB were fed to rats and the ability to induce antigen specific tolerance was tested. LTBSC inhibited the specific delayed-type hypersensitivity (DTH) response and induced decreased antigen-specific in vivo and in vitro cell proliferation more efficiently than LTB. Thus, the novel hybrid molecule LTBSC when orally delivered was able to elicit a systemic immune response. These results suggest that LTBSC could be suitable for exploring further therapeutic treatment of autoimmune inflammatory diseases involving antigens from central nervous system. PMID- 18400514 TI - Expression, purification, refolding and characterization of a putative lysophospholipase from Pyrococcus furiosus: retention of structure and lipase/esterase activity in the presence of water-miscible organic solvents at high temperatures. AB - A putative lysophospholipase (PF0480) encoded by the Pyrococcus furiosus genome has previously been cloned and expressed in Escherichia coli. Studies involving crude extracts established the enzyme to be an esterase; however, owing presumably to its tendency to precipitate into inclusion bodies, purification and characterization have thus far not been reported. Here, we report the overexpression and successful recovery and refolding of the enzyme from inclusion bodies. Dynamic light scattering suggests that the enzyme is a dimer, or trimer, in aqueous solution. Circular dichroism and fluorescence spectroscopy show, respectively, that it has mixed beta/alpha structure and well-buried tryptophan residues. Conformational changes are negligible over the temperature range of 30 80 degrees C, and over the concentration range of 0-50% (v/v) of water mixtures with organic solvents such as methanol, ethanol and acetonitrile. The enzyme is confirmed to be an esterase (hydrolyzing p-NP-acetate and p-NP-butyrate) and also shown to be a lipase (hydrolyzing p-NP-palmitate), with lipolytic activity being overall about 18- to 20-fold lower than esterase activity. Against p-NP-palmitate the enzyme displays optimally activity at pH 7.0 and 70 degrees C. Remarkably, over 50% activity is retained at 70 degrees C in the presence of 25% acetonitrile. The high organic solvent stability and thermal stability suggest that this enzyme may have useful biodiesel-related applications, or applications in the pharmaceutical industry, once yields are optimized. PMID- 18400515 TI - Amorphous calcium carbonate biomineralization in the earthworm's calciferous gland: pathways to the formation of crystalline phases. AB - In this study, we investigated the microstructural transformations that take place during carbonate formation in the earthworm's calciferous gland by analysing the evolution from the precursor fluid of the solid phases (spherulites) to the final carbonate concretions released by the gland. Results from HREM and electron diffraction showed that the spherulithic deposits merely consisted of ACC partially transformed to vaterite. Furthermore, comparisons of the diffraction spectra and microstructural analyses allowed the identification of the transition sequences to more stable carbonates. And thus, transformations of ACC to calcite were observed on the surfaces of these amorphous globular aggregates as their smooth characteristic surface became rougher with time. This transition path was not unique, and the presence of aragonite, as an intermediate phase, has also been found. In this particular case, the transition process followed a completely different pathway with the crystallization starting in the centre of the sphere and progressively extending to the periphery, leading to the formation of radial aggregates. In situ experiments performed on the freshly extracted precursor fluid and analysed by FT-IR spectroscopy showed that ACC is the main constituent and is probably stabilised by macromolecules such as proteins and sugars. Furthermore, the Debye-Scherrer diffraction experiments showed that the carbonate phase present in this fluid remains stable as ACC for more than a week. All these features are indicative of this entire process being biologically controlled by the earthworms. The analysis of the amorphous structure factor of this ACC indicates that these transformations are preceded by short-range order modifications of the amorphous precursor phase. PMID- 18400516 TI - Expression of interferon and interferon-induced genes in salmonids in response to virus infection, interferon-inducing compounds and vaccination. AB - Interferons (IFNs) involved in innate immunity against viruses have recently been cloned from Atlantic salmon and rainbow trout. Moreover, several IFN-stimulated genes (ISGs) have been cloned from salmonids although only Mx has been shown to possess antiviral properties. Much less in known about how viruses induce IFNs in salmonids, but synthetic ligands for some of the main mammalian viral sensors also induce IFNs and ISGs in salmonids. Analysis of the promoters of the salmon IFN-alpha1 and IFN-alpha2 genes shows that activation is dependent on both NFkappaB and IRFs similar to human IFN-beta. Furthermore, several IFN-stimulated genes (ISGs) have been cloned from salmonids although only Mx has been shown to possess antiviral properties. The synthetic compounds poly I:C, imidazoquinolines and CpG oligonucleotides induce IFNs and ISGs in salmonids, probably through the same pathways as in mammals. Salmonid viruses show potent ability to stimulate expression of IFN and ISGs in vivo. Differences between viruses in the ability to stimulate host gene expression are often more evident in cell culture, but more work is needed to pinpoint how salmonid viruses antagonize the IFN system of their host. Finally, existing data suggest that IFNs play a role in the early non specific protection observed after vaccination of salmonids with rhabdoviral DNA vaccines and conventional polyvalent vaccines. PMID- 18400517 TI - The relationship between flexibility and EMG activity pattern of the erector spinae muscles during trunk flexion-extension. AB - BACKGROUND: Movements in the lumbar spine, including flexion and extension are governed by a complex neuromuscular system involving both active and passive units. Several biomechanical and clinical studies have shown the myoelectric activity reduction of the lumbar extensor muscles (flexion-relaxation phenomenon) during lumbar flexion from the upright standing posture. The relationship between flexibility and EMG activity pattern of the erector spinae during dynamic trunk flexion-extension task has not yet been completely discovered. OBJECTIVE: The purpose of this study was to investigate the relationship between general and lumbar spine flexibility and EMG activity pattern of the erector spinae during the trunk flexion-extension task. METHODS: Thirty healthy female college students were recruited in this study. General and lumbar spine flexibilities were measured by toe-touch and modified schober tests, respectively. During trunk flexion-extension, the surface electromyography (EMG) from the lumbar erector spinae muscles as well as flexion angles of the trunk, hip, lumbar spine and lumbar curvature were simultaneously recorded using a digital camera. The angle at which muscle activity diminished during flexion and initiated during extension was determined and subjected to linear regression analysis to detect the relationship between flexibility and EMG activity pattern of the erector spinae during trunk flexion-extension. RESULTS: During flexion, the erector spinae muscles in individuals with higher toe-touch scores were relaxed in larger trunk and hip angles and reactivated earlier during extension according to these angles (P<0.001) while in individuals with higher modified schober scores this muscle group was relaxed later and reactivated sooner in accordance with lumbar angle and curvature (P<0.05). Toe-touch test were significantly correlated with trunk and hip angles while modified schober test showed a significant correlation with lumbar angle and curvature variables. CONCLUSION: The findings of this study indicate that flexibility plays an important role in trunk muscular recruitment pattern and the strategy of the CNS to provide stability. The results reinforce the possible role of flexibility alterations as a contributing factor to the motor control impairments. This study also shows that flexibility changes behavior is not unique among different regions of the body. PMID- 18400518 TI - A framework for in vivo quantification of regional brain folding in premature neonates. AB - This paper describes and compares novel approaches to in vivo 3D measurement of brain surface folding in clinically acquired neonatal MR image data, which allows regional folding evaluation. Most of the current measures of folding are not independent of the area of the surface they are derived from. Therefore, applying them to whole-brain surfaces or subregions of different sizes results in differences which may or may not reflect true differences in folding. We address this problem by proposing new measures to quantify gyrification and two approaches to normalize previously defined measures. The method was applied to twelve premature infants (age 28-37 weeks) from which cerebrospinal fluid/gray matter and gray matter/white matter interface surfaces were extracted. Experimental results show that previous folding measures are sensitive to the area of the surface of analysis and that the area-independent measures proposed here provide significant improvements. Such a system provides a tool that facilitates the study of structural development in the neonatal brain within specific functional subregions, which may be critical in identifying later neurological impairment. PMID- 18400520 TI - Investigation of spatial resolution, partial volume effects and smoothing in functional MRI using artificial 3D time series. AB - This work addresses the balance between temporal signal-to-noise ratio (tSNR) and partial volume effects (PVE) in functional magnetic resonance imaging (fMRI) and investigates the impact of the choice of spatial resolution and smoothing. In fMRI, since physiological time courses are monitored, tSNR is of greater importance than image SNR. Improving SNR by an increase in voxel volume may be of negligible benefit when physiological fluctuations dominate the noise. Furthermore, at large voxel volumes, PVE are more pronounced, leading to an overall loss in performance. Artificial fMRI time series, based on high resolution anatomical data, were used to simulate BOLD activation in a controlled manner. The performance was subsequently quantified as a measure of how well the resulted activation matched the simulated activation. The performance was highly dependent on the spatial resolution. At high contrast-to-noise ratio (CNR), the optimal voxel volume was small, i.e. in the region of 2(3) mm(3). It was also shown that using a substantially larger voxel volume in this case could potentially negate the CNR benefits. The optimal smoothing kernel width was dependent on the CNR, being larger at poor CNR. At CNR >1, little or no smoothing proved advantageous. The use of artificial time series gave an opportunity to quantitatively investigate the effects of partial volume and smoothing in single subject fMRI. It was shown that a proper choice of spatial resolution and smoothing kernel width is important for fMRI performance. PMID- 18400519 TI - Structural and functional biomarkers of prodromal Alzheimer's disease: a high dimensional pattern classification study. AB - This work builds upon previous studies that reported high sensitivity and specificity in classifying individuals with mild cognitive impairment (MCI), which is often a prodromal phase of Alzheimer's disease (AD), via pattern classification of MRI scans. The current study integrates MRI and PET (15)O water scans from 30 participants in the Baltimore Longitudinal Study of Aging, and tests the hypothesis that joint evaluation of structure and function can yield higher classification accuracy than either alone. Classification rates of up to 100% accuracy were achieved via leave-one-out cross-validation, whereas conservative estimates of generalization performance in new scans, evaluated via bagging cross-validation, yielded an area under the receiver operating characteristic (ROC) curve equal to 0.978 (97.8%), indicating excellent diagnostic accuracy. Spatial maps of regions determined to contribute the most to the classification implicated many temporal, prefrontal, orbitofrontal, and parietal regions. Detecting complex patterns of brain abnormality in early stages of cognitive impairment has pivotal importance for the detection and management of AD. PMID- 18400521 TI - Global average gray and white matter N-acetylaspartate concentration in the human brain. AB - Since the amino acid derivative N-acetylaspartate (NAA) is almost exclusive to neuronal cells in the adult mammalian brain and its concentration has shown local (or global) abnormalities in most focal (or diffuse) neurological diseases, it is considered a specific neuronal marker. Yet despite its biological and clinical prominence, the relative NAA concentration in the gray and white matter (GM, WM) remains controversial, with each reported to be higher than, equal to, or less than the other. To help resolve the controversy and importantly, access the NAA in both compartments in their entirety, we introduce a new approach to distinguish and quantify the whole-brain average GM and WM NAA concentration by integrating MR-image segmentation, localized and non-localized quantitative (1)H MRS. We demonstrate and validate the method in ten healthy volunteers (5 women) 27+/-6 years old (mean+/-standard-deviation) at 1.5T. The results show that the healthy adult human brain comprises significantly less WM, 39+/-3%, than GM 60+/ 4% by volume (p<0.01). Furthermore, the average NAA concentration in the WM, 9.5+/-1.0 mM, is significantly lower than in GM, 14.3+/-1.1 mM (p<0.01). PMID- 18400522 TI - Population structure and genetic diversity in two species of Hawaiian picture winged Drosophila. AB - Over the last several decades many picture-winged Drosophila have become less common in both geographical distribution and local population size (pers. obs., Foote pers. comm., Montgomerey pers. comm.). Here we report on a study of two Hawaiian Drosophila species, D. engyochracea, and D. hawaiiensis, to determine the impact that changes in population sizes over the past thirty years have had on the genetic diversity of these species. D. engyochracea is known from only two locations on the Island of Hawai'i (Kipuka Ki and Kipuka Pua'ulu), while D. hawaiiensis is currently more wide spread across Hawai'i Island. We collected 65 D. hawaiiensis and 66 D. engyochracea from two forest patches (kipuka) isolated by a 400 year old volcanic ash deposit. DNA sequence data for 515 bases of the mitochondrial gene COII was analyzed for both species to estimate relative total genetic diversity as well as inter-kipuka gene flow. The more wide spread species, D. hawaiiensis, has more genetic diversity (23 vs. 11 unique haplotypes) than the rarer species, D. engyochracea. The distribution of haplotypes in the kipuka is consistent with more gene flow in D. engyochracea than in D. hawaiiensis. Phylogenetic analysis indicates a small number of individuals morphologically identified as one species but have DNA sequence diagnostic for the other species. These results are consistent with these individuals being descendant from hybrids between species. PMID- 18400523 TI - Acute hepatic injury in a child with Dravet syndrome: no protective effect of stiripentol. PMID- 18400524 TI - Status epilepticus in epileptic patients. Related syndromes, precipitating factors, treatment and outcome in a video-EEG population-based study. AB - INTRODUCTION: Status epilepticus (SE) is frequently observed in epileptic patients. We reviewed a series of video-EEG documented SE to define the characteristics of SE in this population. MATERIALS AND METHODS: Retrospective evaluation of 50 epileptic patients with SE, revision of the electro-clinical data and therapies, and definition of the semeiological subtypes, aetiology, outcome and related epileptic syndromes. RESULTS: We identified 28 convulsive (19 focal and 9 generalized) and 22 non-convulsive (8 focal and 14 generalized) SE patients. In 13 patients, SE was situation-related (poor compliance, AED reduction, worsening seizures). In the remaining 37 patients, SE was related to the natural history of epilepsy (progression of underlying pathologies or intrinsic expression of epileptic syndromes); in these last cases, our results show a higher occurrence in cryptogenic frontal epilepsy (p=0.01). We identified two subgroups according to the duration of the event, i.e. SE lasting <12h and SE lasting >12h. Our results showed a worse response to therapy in SE lasting >12h (p=0.01), a better response to therapy in non-convulsive SE than in convulsive SE (p<0.05) and a relationship at statistical significance limit between a poor response to therapy/worse outcome and symptomatic epileptic syndromes (p=0.06). CONCLUSION: SE in epileptic patients has a wide spectrum of electro-clinical features. It may be related to the withdrawal or reduction of AEDs, or may even be the expression of the evolution of epileptic syndromes. Response to therapy is dependent on early diagnosis and therapy. PMID- 18400525 TI - Articular chondrocytes derived from distinct tissue zones differentially respond to in vitro oscillatory tensile loading. AB - OBJECTIVE: The cell morphology, gene expression, and matrix synthesis of articular chondrocytes are known to vary with depth from the tissue surface. The objective of this study was to investigate if chondrocytes from different zones respond to in vitro oscillatory tensile loading in distinct ways and whether tensile strain, which is most prevalent near the articular surface, would preferentially stimulate superficial zone chondrocytes. DESIGN: Chondrocytes were separately isolated from the superficial, middle, and deep zones of articular cartilage and seeded into three-dimensional fibrin hydrogel constructs. An intermittent protocol of oscillatory tensile loading was applied for 3 days, and the effects on extracellular matrix (ECM) synthesis were assessed by measuring the incorporation of radiolabed precursors, size exclusion gel chromatography, and western blotting. RESULTS: Tensile loading was found to be a potent stimulus for proteoglycan synthesis only in superficial zone chondrocytes. Although overall biosynthesis rates by deep zone chondrocytes were unaffected by tensile loading, the molecular characteristics of proteins and proteoglycans released to the culture medium were significantly altered so as to resemble those of superficial zone chondrocytes. CONCLUSIONS: Oscillatory tensile loading differentially affected subpopulations of articular chondrocytes in three dimensional fibrin hydrogel constructs. Cells isolated from deeper regions of the tissue developed some characteristics of superficial zone chondrocytes after exposure to tensile loading, which may indicate an adaptive response to the new mechanical environment. Understanding how exogenous mechanical stimuli can differentially influence chondrocytes from distinct tissue zones will yield important insights into mechanobiological processes involved in cartilage tissue development, maintenance, disease, and repair. PMID- 18400526 TI - Comparison between osteoblasts derived from human dental pulp stem cells and osteosarcoma cell lines. AB - Stem cells derived from human dental pulp are able to differentiate into osteoblasts and are a potential source of autologous bone. The aim of this study was to compare genes differentially expressed in osteoblastoids from human dental pulp (OHDP) to osteosarcoma cells (OCs). Human dental pulp was extracted and immersed in a digestive solution. Cells were cultured and selected using c-kit, CD34, CD45 and STRO-1 antibodies. In parallel, two OCs (i.e., SAOS2 and TE85) were cultured. RNA was extracted from different populations of cells and cDNA was used for the hybridisation of human 19.2K DNA microarrays. We identified several differences in gene expression between OHDP and OCs. Some down-regulated OHDP genes, such as RUNX1, MAP4K4 and PRDM2, are involved in bone development, cell motility and transcript regulation. Gene expression in OHDP is significantly different from that in OCs, suggesting differences in cell function and activity between these cells. PMID- 18400527 TI - The mechanism of myoblast deformation in response to cyclic strain - A cytomechanical study. AB - Mechanical strain is one of the important epigenetic factors that cause deformation and differentiation of skeletal muscles. This research was designed to investigate how myoblast deformation occurs after cyclic strain loading. Myoblasts were passaged three times and harvested; various cyclic strains (2.5kPa, 5kPa and 10kPa) were then loaded using a pulsatile mechanical system. The adaptive response of the myoblasts was observed at different time points (0.5h, 1h, 6h and 12h) post-loading. At the early stage of cyclic strain loading (<1h), almost no visible morphological changes were observed in the myoblasts. The actin cytoskeleton showed a disordered arrangement and a weak fluorescence expression; there was little expression of talin. At 6h and 12h post-loading, the myoblasts changed their orientation to parallel (in the 2.5kPa and 5kPa groups) or perpendicular (in the 10kPa group) to the direction of strain. Fluorescence expression of both the actin cytoskeleton and talin was significantly increased. The results suggest that cyclic strain has at least two ways to regulate adaptation of myoblasts: (1) by directly affecting actin cytoskeleton at an early stage post-loading to cause depolymerization; and (2) by later chemical signals transmitted from the extracellular side to intracellular side to initiate repolymerization. PMID- 18400528 TI - Invited commentary on: separation of components in fenestrated and branched endovascular grafting--branch protection or a potentially new mode of failure? PMID- 18400529 TI - These DisUnited States of Health Care: a call for improved patient convenience in access to oral and maxillofacial radiology services. PMID- 18400530 TI - Congenital heart disease in boxer dogs: results of 6 years of breed screening. AB - The aim of this study was to analyse the results of 6years (1999-2004) of mandatory breed screening for congenital heart disease in Boxer dogs using physical examination and echocardiography. Records of 1283 Boxers were reviewed and 165 dogs (12.86%) were found to be affected by heart disease, with aortic and pulmonic stenosis being the most frequent cardiac lesions. Comparison of these results with those of a previous survey showed a lower overall prevalence of both outflow obstructions, particularly of the more severe forms. A male predisposition for both aortic and pulmonic stenosis was evident from the study. Consistent with reports from other countries, soft left basilar heart murmurs were detected in both healthy dogs and dogs affected with congenital heart disease. PMID- 18400531 TI - Ultrasonographic characteristics (cross-sectional area and relative echogenicity) of the digital flexor tendons and ligaments of the metacarpal region in Purebred Spanish horses. AB - The objectives of this study were to establish normal values for relative echogenicity (RE) and cross-sectional area (CSA) of the flexor tendons and ligaments of the metacarpal region of Purebred Spanish horses (PBSH), and to determine the effect of body mass index (BMI), age, sex and forelimb on these variables. Transverse ultrasonographic images were obtained using 20 normal PBSH (10 females, 10 males; 2-25 years of age; BMI 132.04-152.25; not in training). The images were digitised and values for CSA and RE were determined for each structure. The suspensory ligament was the most echogenic and the largest structure, and the superficial digital flexor tendon was the least echogenic and smallest of the tissues examined. The tendons and ligaments of the metacarpal region in PBSH showed similar RE but smaller CSAs when compared to other breeds. PMID- 18400532 TI - Controlling the equine neutrophil: a generator of devastating tissue damage during equine inflammatory disease. PMID- 18400533 TI - Molecular modeling study of CodX reveals importance of N-terminal and C-terminal domain in the CodWX complex structure of Bacillus subtilis. AB - In Bacillus subtilis, CodW peptidase and CodX ATPase function together as a distinctive ATP-dependent protease called CodWX, which participates in protein degradation and regulates cell division. The molecular structure of CodX and the assembly structure of CodW-CodX have not yet been resolved. Here we present the first three-dimensional structure of CodX N-terminal (N) and C-terminal (C) domain including possible structure of intermediate (I) domain based on the crystal structure of homologous Escherichia coli HslU ATPase. Moreover, the biologically relevant CodWX (W(6)W(6)X(6)) octadecamer complex structure was constructed using the recently identified CodW-HslU hybrid crystal structure. Molecular dynamics (MD) simulation shows a reasonably stable structure of modeled CodWX and explicit behavior of key segments in CodX N and C domain: nucleotide binding residues, GYVG pore motif and CodW-CodX interface. Predicted structure of the possible I domain is flexible in nature with highly coiled hydrophobic region (M153-M206) that could favor substrate binding and entry. Electrostatic surface potential observation unveiled charge complementarity based CodW-CodX interaction pattern could be a possible native interaction pattern in the interface of CodWX. CodX GYVG pore motif structural features, flexible nature of glycine (G92 and G95) residues and aromatic ring conformation preserved Y93 indicated that it may follow the similar mode during the proteolysis mechanism as in the HslU closed state. This molecular modeling study uncovers the significance of CodX N and C domain in CodWX complex and provides possible explanations which would be helpful to understand the CodWX-dependent proteolysis mechanism of B. subtilis. PMID- 18400534 TI - Insect transferrin functions as an antioxidant protein in a beetle larva. AB - In insects transferrin is known as an iron transporter, an antibiotic agent, a vitellogenin, and a juvenile hormone regulated protein. Here, a novel functional role for insect transferrin as an antioxidant protein is demonstrated. Stressors, such as heat shock, fungal challenge, and H(2)O(2) exposure, cause upregulation of the white-spotted flower chafer Protaetia brevitarsis (Coleoptera: Scarabaeidae) transferrin (PbTf) mRNA in the fat body and increases PbTf protein levels in the hemolymph. RNA interference (RNAi) treated PbTf reduction causes increased iron and H(2)O(2) levels in the hemolymph and results in induction of apoptotic cell death in the fat body during exposure to stress. The observed effects of PbTf RNAi suggest that PbTf inhibits stress-induced apoptosis by diminishing the Fenton reaction via the binding of iron, thus supporting an antioxidant role for PbTf in stress responses. PMID- 18400535 TI - A non-synonymous variant in SLC30A8 is not associated with type 1 diabetes in the Danish population. AB - Genome-wide association scans in type 2 diabetes (T2D) have identified a risk variant, rs13266634 (Arg325Trp), in SLC30A8 on chromosome 8. SLC30A8 encodes a beta-cell specific zinc-ion transporter and rs13266634 has been shown to affect insulin secretion. Recently, autoantibodies for Slc30A8 with high predictive value were demonstrated in individuals with type 1 diabetes (T1D), making this gene an interesting T1D candidate gene. We genotyped rs13266634 in 3008 cases and controls and 246 families from Denmark. Association to T1D could not be demonstrated. PMID- 18400536 TI - Bartonella species antibodies and DNA in cerebral spinal fluid of cats with central nervous system disease. AB - Bartonella species infection is associated with central nervous system (CNS) disease in some humans and cats but the diagnosis is difficult to confirm with blood or serum test results. In this retrospective study of 100 client-owned cats, serum and cerebral spinal fluid (CSF) were assayed for Bartonella species IgG antibodies and CSF was assayed for Bartonella species DNA. Bartonella species IgG antibodies were detected in serum of 36 cats, Bartonella species C-values>1 (suggesting antibody production by the CNS) were detected in CSF of 11 cats, and B henselae DNA was amplified from the CSF of 10 cats. While the clinical significance of these findings cannot be assessed without a control group, the development of neurological signs in some cats inoculated with B henselae and the results of this study warrant prospective evaluation of the association of Bartonella species with feline CNS disease. PMID- 18400537 TI - De novo N-palmitoylsphingosine synthesis is the major biochemical mechanism of ceramide accumulation following p53 up-regulation. AB - The tumor suppressor protein p53 and the putative lipid tumor suppressor ceramide play pivotal roles in inducing cell cycle arrest or in driving the cell towards apoptosis. Previously we had shown that, in a p53-dependent model of cell death, ceramide accumulated in a p53-dependent manner [Dbaibo GS, Pushkareva MY, Rachid RA, Alter N, Smyth MJ, Obeid LM, Hannun YA. J Clin Invest 1998;102:329-339]. In the current study, we investigated the biochemical pathways by which ceramide accumulated following p53 up-regulation. In both Molt-4 LXSN leukemia cells exposed to gamma-irradiation and in EB-1 colon cancer cells treated with ZnCl(2), p53 up-regulation led to de novo ceramide synthesis with predominance of N palmitoylsphingosine (C16-ceramide) synthesis. The activation of the de novo pathway was not associated with increased activity of the key enzyme serine palmitoyltransferase (SPT) but rather with the increased activity of ceramide synthase. Furthermore, transcriptional up-regulation of the palmitoyl-specific Lass5 ceramide synthase gene was observed in Molt-4 but not in EB-1 cells. The SPT inhibitor ISP-1 or the ceramide synthase inhibitor fumonisin B1 led to substantial inhibition of ceramide accumulation in response to p53 up-regulation. Other biochemical pathways of ceramide generation such as sphingomyelinase activation were examined and found unlikely to contribute to p53-dependent ceramide formation. These studies indicate that p53 specifically drives de novo ceramide synthesis by activation of a ceramide synthase that favors the synthesis of N-palmitoylsphingosine. PMID- 18400538 TI - Modulation of total ceramide and constituent ceramide species in the acutely and chronically hypoxic mouse heart at different ages. AB - Ceramide has been implicated in regulatory processes vital for cell survival under different stressors, most notably hypoxia. Little has been done to investigate the contributions of the different ceramide species to the regulation of cell survival. This study aims to highlight the patterns of variation in total ceramide and its species in the growing and hypoxic mouse heart. Mus musculus mice were placed in a hypoxic environment at birth. Control animals remained in room air. The hearts were extracted at different time points: 1 day, 1 week, 4 weeks, and 8 weeks. The total ceramide content and the amounts of component species were assayed by a modified diacylglycerol kinase assay and high performance liquid chromatography-tandem mass spectroscopy, respectively. Data was collected from both ventricles in hypoxic and control conditions. There was significant polycythemia in the hypoxic versus control animals with a nearly twofold increase in hematocrit levels. Hypoxic right ventricle (RV) mass significantly increased over that of controls at different age groups. When ceramide content was compared in the hypoxic versus control animals, there was a significant increase at day 1 and a significant decrease at week 4 in the left ventricle, whereas a significant decrease was found in the RV at 1 week, 4 weeks, and 8 weeks. There was also a differential involvement of the RV with regard to levels of N-palmitoyl-D-erythro-sphingosine (C16-Cer) and its synthetic precursor dihydro-N-palmitoyl-D-erythro-sphinganine (DHC-16-Cer). The decrease in C16-Cer observed in both hypoxic and control RV's over time was paralleled by a significant increase in DHC-16-Cer in hypoxic (142.1+/-15.0 pmol; p<0.05) but not control (52.8+/-4.0 pmol) RV's suggesting a role for DHC-16-Cer in the RV adaptive response to hypoxia. Another species, N-arachidoyl-D-erythro-sphingosine (C20-Cer), was specifically and significantly decreased in the hypoxic RV. These studies support the presence of distinct roles for different ceramide species and their precursors. A better assessment of cyanotic congenital heart disease in light of the mechanism and timing of cardiomyocyte death, will lead to punctual interventions and even novel cardioprotective strategies. PMID- 18400539 TI - Prevalence of hepatitis B virus markers among blood donors in a tertiary hospital in Tabuk, northwestern Saudi Arabia. AB - OBJECTIVE: This study was carried out to determine the prevalence of hepatitis B virus (HBV) markers among blood donors in a tertiary hospital in Tabuk, northwestern Saudi Arabia. MATERIALS AND METHODS: A cross-sectional survey was done among male Saudi voluntary blood donors at the blood bank of the King Khalid tertiary care hospital in the northwest region of Saudi Arabia. A total of 3192 subjects were included in the study over the period from June 1, 2005 to May 31, 2006. Subjects were tested for evidence of HBV infection plus other relevant blood-borne pathogens. A questionnaire sheet for the collection of personal information (such as name, age, and sex), socioeconomic data (such as marital status, education, and occupation), family history of HBV infection, history of immunization against HBV, and history of past exposure to a high risk procedure or behavior, was prepared and used for each subject. Data were tabulated and statistically analyzed. RESULTS: Among the studied sample, the prevalence of HBsAg (hepatitis B surface antigen) was found to be 3.0% and the prevalence of anti-HBc (anti-hepatitis B core antigen) was found to be 18.7%. Blood donors with positive HBV markers showed significant association with increased age, being married, lower educational level, specific occupations such as blue collar worker and the military, family history of HBV infection, and lack of immunization. However, there was no significant association with history of exposure to high risk procedure or behavior. CONCLUSIONS: The prevalence of HBV infection among male Saudi blood donors in the northwest region of Saudi Arabia is high compared to that reported from the central capital area (Riyadh). Public health education programs on HBV infection, adult hepatitis B immunization programs, and raising socioeconomic standards should be initiated in order to target the high-risk groups found in this study. Furthermore, a community-based study is recommended to investigate HBV infection in the northwest region of Saudi Arabia. PMID- 18400540 TI - Co-existent pneumococcal and tubercular mixed meningitis in heterozygous sickle cell disease: a case report. PMID- 18400541 TI - Is phage therapy acceptable in the immunocompromised host? AB - Over the last decade, bacteriophages (bacterial viruses) have emerged as the major alternative to antibiotics in the treatment of antibiotic-resistant infections. While a considerable body of evidence has accumulated for the efficacy and safety of phage therapy in immunocompetent patients, data remain relatively scarce regarding its use in the immunocompromised host. To our knowledge, the present article is the first to summarize all findings, of both experimental and clinical studies, that may be relevant to the employment of phage therapy in immunocompromised patients. The available data suggest that bacteriophages could also be an efficacious and safe therapeutic modality in such patients. PMID- 18400542 TI - The seroepidemiology of Helicobacter pylori infection in Australia. AB - BACKGROUND: Infection with Helicobacter pylori is common worldwide and a significant cause of upper gastrointestinal disease. Prevalence of this infection varies in different population groups internationally. Because of the invasiveness of specimen collection for bacteriologic diagnosis and the expense of tests such as labeled urea breath tests, serology is the most feasible means of determining the population epidemiology of H. pylori. The aim of this study was to describe the seroepidemiology of H. pylori infection in Australia. METHODS: H. pylori-specific ELISA for the presence of IgG antibodies was performed on a representative sample of 2413 sera from Australia in 2002, using validated serosurveillance methods. RESULTS: The overall seroprevalence of H. pylori infection in Australia was 15.1% in 2002, with no statistical difference between genders. Seropositivity rates increased progressively with age, ranging between 4.0% in the 1-4-year-olds and 23.3% in the 50-59-year-olds. CONCLUSIONS: The prevalence of infection with H. pylori in Australia was lower than rates reported in other developed countries, at 15.4%. This study provides important baseline measurements for future preventive measures including vaccine research and development. Further studies to determine subgroups at higher risk of infection may help target the more susceptible populations. PMID- 18400543 TI - Actinomycosis of the appendix: diagnostic and therapeutic considerations. PMID- 18400544 TI - The antibacterial activity against MRSA strains and other bacteria of a <500Da fraction from maggot excretions/secretions of Lucilia sericata (Diptera: Calliphoridae). AB - The application of Lucilia sericata larvae to chronic, infected wounds results in the rapid elimination of infecting microorganisms, including MRSA. Previously, we demonstrated in vitro antibacterial activity of native excretions/secretions (nES) from L. sericata and partially purified two low mass antibacterial compounds with masses of 0.5-10kDa and <500Da. The present study reports the antibacterial effects of the <500Da fraction (ES<500) on the growth and morphology of a range of bacteria, including 12 MRSA strains. Distinct morphological changes were observed in Bacillus cereus and Escherichia coli following exposure to ES<500. Flow cytometry and confocal microscopy analyses, in conjunction with turbidometric and CFU assays, revealed bacteriostatic activity of nES against S. aureus and E. coli. ES<500 also demonstrated bacteriostatic activity against S. aureus, however, bactericidal activity and the induction of a viable but non-culturable state were observed with ES<500-treated E. coli. PMID- 18400545 TI - Rosetting is associated with increased Plasmodium falciparum in vivo multiplication rate in the Saimiri sciureus monkey. AB - Severe Plasmodium falciparum malaria in African children is associated with high peripheral parasite densities and high rate of rosette-forming parasites. To explore the relationship between rosette formation and parasite density in vivo, we compared the multiplication rate of a rosette-forming variant (varO) of the Palo Alto line with a sibling non-rosetting variant (varR) in splenectomized Saimiri monkeys. The multiplication rate of varO parasites was 1.5-fold higher than that of the varR variant. This indicates that rosetting is indeed associated with high parasite multiplication efficiency in vivo and, as such, may contribute to the high parasite densities observed in severe malaria. PMID- 18400547 TI - The Moraxella IgD-binding protein MID/Hag is an oligomeric autotransporter. AB - The immunoglobulin D (IgD)-binding protein MID/Hag of the human respiratory pathogen Moraxella catarrhalis is an outer membrane protein of approximately 200kDa belonging to the autotransporter family. MID also functions as an adhesin and hemagglutinin. In the present paper, the ultrastructure of MID was mapped. Using a series of Escherichia coli transformants, the last 210 aa of the C terminal region were shown to translocate protein MID through the outer membrane suggesting that MID has a beta-barrel structure comprising of 10 transmembrane beta-sheets. Electron microscopy mapping with gold-labelled specific antibodies, and partial unravelling using guanidine hydrochloride showed that the rest of the MID protein forms an approximately 120nm long, fibrillar structure in which the individual monomers fold back on themselves to expose a globular distal domain at their tips comprising both the IgD-binding (MID962-1200) and adhesive (MID764 913) regions. This positions their N-termini close to the C-terminal membrane spanning domains. Mass measurements by scanning transmission electron microscopy (STEM) verified that the MID molecule is an oligomer. PMID- 18400546 TI - Decreased inflammatory response in Toll-like receptor 2 knockout mice is associated with exacerbated Pneumocystis pneumonia. AB - Pneumocystis pneumonia (PcP) is marked by substantial inflammatory damage to the lung. We have found that Toll-like receptor 2 (TLR2) mediates macrophage inflammatory responses to Pneumocystis and hypothesized that TLR2 deficiency would lead to less severe inflammation and milder lung injury during PcP. Histopathology examination showed that TLR2-/- mice with PcP indeed exhibited milder pulmonary inflammation. TLR2-/- mouse lungs contained less TNF-alpha and displayed lower levels of NF-kappaB activation during PcP. However, TLR2-/- mice with PcP displayed increased severity in symptoms and organism burden. The increased organism burden is likely due to defects in protective mechanisms in TLR2-/- mice. mRNA levels of the inducible nitric oxide synthase and NADPH oxidase p47phox, as well as nitric oxide levels in the lungs, were decreased in TLR2-/- PcP mice. Taken together, this study shows that TLR2-mediated inflammatory responses contribute to a certain degree to the clearance of Pneumocystis organism in mice. PMID- 18400548 TI - [A study comparing previous induced abortion rates in populations of newly delivered women and infertile women]. AB - OBJECTIVE: For years, induced abortions (IA) have been deemed responsible for altered fertility. The implication of various mechanisms including tubal infertility, intra-uterine adhesions, spontaneous abortion, ectopic pregnancy, cervical incompetence, shortened gestations, and any psychological trouble leading to anovulation has been raised. Though many authors find no evidence of an increased risk on fertility for women, whose IA is not complicated by infection, it might sometimes be insinuated that infertility is the consequence of previous abortion. Thus, we compared the rate of patients with any prior IA in a population of newly delivered women and in women ongoing IVF. PATIENTS AND METHODS: A retrospective study was conducted at Sevres hospital, comparing two populations of women, newly delivered women (n=1738) between January 1st and December 31st 2005, and women ongoing IVF at the same period (n=430). We reported the number of previous medical or surgical induced abortions and compared it in both groups. Another analysis compared these rates, among two subgroups of women with one or more prior pregnancy (secondary infertility (n=148), and secondary gestation (n=1088). RESULTS: The rate of prior IA was not different in the two populations. In the IVF group, 13% (n=56) had undergone one or more IA, versus 16.7% (n=291) in the newly delivered group (P=0.06). Among women with previous pregnancy, 37.8% (56) women of the IVF group had undergone one or more previous IA, versus 26.7% (291) of the newly delivered women (P=0.007). DISCUSSION AND CONCLUSION: As expected by literature data on IA and fertility, rates of prior induced abortions were not different in the population of fertile women versus infertile. However, women with one or more previous pregnancy are more likely to have undergone previous IA in the IVF group than in the newly delivered group, possibly due to a bias of age. More data are requested to eliminate linkage between IA and infertility. PMID- 18400549 TI - Validity of the local nonlinear arterial flow theory: influence of the upstream and downstream conditions. AB - The "local flow" theory provides a simple way to take into account the nonlinear convective terms associated with blood flow in large arteries. The assumption that blood velocity profiles vary slowly with the longitudinal coordinate allows a simple nonlinear resolution via a mathematical approximation. Although validated in vitro by its authors, this theory still needs to be verified in accordance with the range of variation of the hemodynamical parameters. This constitutes the aim of this work where we assess the validity of two models issued from this theory: the "direct model" using the pressure-gradient as an input for the calculations and the "indirect model" using the centre-line velocity. The assessment of these models is made by comparing their solutions to those of the linear theory using numerical simulations. Our main conclusion is that the indirect model has a wide range of validity while the direct one fails in the presence of a strong reflected wave. PMID- 18400550 TI - The effect of the subcutaneous fat on the transfer of current through skin and into muscle. AB - The present investigation was conducted to see the effect of subcutaneous fat on the transmission characteristics of an electrical stimulus applied to the skin and conducted to skeletal muscle. Two groups of subjects participated. In one, the subjects were three males and three females whose average age was 24.6+/-1.5 years, average weight was 74.8+/-18.2kg, and average height was 176.4+/-10.3cm. The other was a group of 30 subjects who average age was 26.2+/-1.9 years, average height 177.3+/-11.5cm, average weight 92.4+/-19.8kg. Electrical stimulation was applied above the quadriceps muscle at a current of 5mA and with sine and square wave stimulation at a frequency of 30Hz and a pulse width of 250micros. Current movement was measured on the skin and into muscle with surface and needle electrodes. The results showed that the thickness of the subcutaneous fat layer was directly related to signal loss from the skin (correlation between subcutaneous fat thickness and RC time constant was 0.96, p<0.001). Because of the subcutaneous fat layer and the resulting capacitance, an RC low pass filter is created such that square wave stimuli are not transmitted well into muscle whereas sine wave stimuli pass easily. Thus, when considering surface stimulation of nerve or muscle, any volume conductor model must take subcutaneous fat into consideration since the RC low pass filter created by fat will filter surface signals or, conversely, signals such as EMG which are generated in muscle but measured on the surface of the skin. PMID- 18400551 TI - Rate and accuracy of bacterial protein synthesis revisited. AB - Our understanding of the accuracy of tRNA selection on the messenger RNA programmed ribosome has recently increased dramatically because of high resolution crystal structures of the ribosome, cryo-electron microscopy reconstructions of its functional complexes, and fast kinetics experiments. Application of single-molecule spectroscopy with fluorescence resonance energy transfer to studies of tRNA selection by the ribosome has also provided new, albeit controversial, insights. Interestingly, when the fundamental trade-off between rate and accuracy in substrate-selective biosynthetic reactions is taken into account, some aspects of the current models of ribosome function appear strikingly suboptimal in the context of growing bacterial cells. PMID- 18400553 TI - Photoregulation in prokaryotes. AB - The spectroscopic identification of sensory rhodopsin I by Bogomolni and Spudich in 1982 provided a molecular link between the light environment and phototaxis in Halobacterium salinarum, and thus laid the foundation for the study of signal transducing photosensors in prokaryotes. In recent years, a number of new prokaryotic photosensory receptors have been discovered across a broad range of taxa, including dozens in chemotrophic species. Among these photoreceptors are new classes of rhodopsins, BLUF-domain proteins, bacteriophytochromes, cryptochromes, and LOV-family photosensors. Genetic and biochemical analyses of these receptors have demonstrated that they can regulate processes ranging from photosynthetic pigment biosynthesis to virulence. PMID- 18400552 TI - The elusive object of desire--interactions of bacteriophages and their hosts. AB - Bacteria and their viruses (phages) are locked in an evolutionary contest, with each side producing constantly changing mechanisms of attack and defense that are aimed to increase the odds of survival. As a result, phages play central roles in a great variety of genetic processes and increase the rate of evolutionary change of the bacterial host, which could ultimately work to the benefit of the host in a long run. PMID- 18400554 TI - Spectroscopic and structural elucidation of merocyanine dye 2,5-[1-metyl-4-[2-(4 hydroxyphenyl)ethenyl)]piridinium]-hexane tetraphenylborate aggregation processes. AB - Structural and spectroscopic elucidation of merocyanine dye, 2,5-[1-metyl-4-[2-(4 hydroxyphenyl)ethenyl)]piridinium]-hexane tetraphenylborate, is performed in gas and condense phase by means of solution and solid-state conventional and linear polarized IR-spectroscopy of oriented colloids in nematic liquid crystal suspension, UV-vis and fluorescence methods, HPLC MS/MS tandem and ESI mass spectrometry, (1)H, (13)C and (1)H-(1)H COSY NMR, TGV and DSC methods. Quantum chemical DFT calculations are performed for structural optimization and spectroscopic properties prediction. PMID- 18400555 TI - Increased biomagnetic activity in the ventral pathway in mild cognitive impairment. AB - OBJECTIVE: Mild cognitive impairment (MCI) patients represent an intermediary state between healthy aging and dementia. MCI activation profiles, recorded during a memory task, have been studied either through high spatial resolution or high temporal resolution techniques. However, little is known about the benefit of combining both dimensions. Here, we investigate, by means of magnetoencephalography (MEG), whether spatio-temporal profiles of neuromagnetic activity could differentiate between MCI and age-matched elderly participants. METHODS: Taking the advantage of the high temporal resolution and good spatial resolution of MEG, neuromagnetic activity from 15 elderly MCI patients and 20 age matched controls was recorded during the performance of a modified version of the Sternberg paradigm. RESULTS: Behavioral performance was similar in both groups. A between group analysis revealed that MCI patients showed bilateral higher activity in the ventral pathway, in both the target and the non-target stimuli. A within-group analysis of the target stimuli, indicates a lack of asymmetry through all late latency windows in both groups. CONCLUSIONS: MCI patients showed a compensatory mechanism represented by an increased bilateral activity of the ventral pathway in order to achieve a behavioral performance similar to the control group. SIGNIFICANCE: This spatio-temporal pattern of activity could be another tool to differentiate between healthy aging and MCI patients. PMID- 18400556 TI - Comparative assessment of best conventional with best theta burst repetitive transcranial magnetic stimulation protocols on human motor cortex excitability. AB - OBJECTIVE: Comparative assessment of best conventional with best theta burst repetitive transcranial magnetic stimulation (rTMS) protocols on human motor cortex excitability. METHODS: Stimulation of dominant hand abductor digiti minimi muscle representation of motor cortex in nine healthy right-handed subjects utilising two different MagPro stimulators to generate TMS pulses of distinct monophasic, half sine and biphasic shape with anteriorly or posteriorly directed current flow, measuring resting and active motor threshold and motor-evoked potential (MEP) amplitude. Comparison of two different conventional rTMSs and eight different theta burst stimulation (TBS) protocols. RESULTS: Continuous TBS (cTBS) yielded an inhibition, and intermittent TBS (iTBS) a facilitation of MEP amplitudes. These effects were not significantly influenced by waveform or current orientation. Compared to conventional rTMS paradigms, neither cTBS nor iTBS showed significantly stronger effects. In addition and confirming earlier findings, the asymmetric monophasic pulse and a posteriorly directed current flow yielded a higher threshold than all other TMS types. The less asymmetric half sine pulse had a lower threshold for postero-anterior current than in the opposite direction. The biphasic stimuli yielded the lowest motor threshold particularly with an antero-posterior current direction. CONCLUSIONS: TBS did not depend on current direction or on pulse configuration in this sample of healthy subjects. In addition, TBS effect size was not clearly superior to conventional rTMS protocols. SIGNIFICANCE: This systematic comparison of three waveforms, two current directions and two different interventional protocols allows a better understanding of the mechanisms of TMS on human motor cortex. PMID- 18400557 TI - Beneficial effects of bisoprolol on the survival of hypertensive diastolic heart failure model rats. AB - BACKGROUND: beta-blocker therapy is an established therapeutic strategy for systolic heart failure. However, its benefits in diastolic heart failure (DHF) are controversial. AIMS: This study was designed to investigate the effects of bisoprolol on DHF. METHODS AND RESULTS: Dahl salt-sensitive rats fed on 8% NaCl diet from age 6 weeks, DHF model rats, were divided into three groups at age 13 weeks. One group was treated with bisoprolol 12.5 mg/kg/day (Low dose group, n=18), one group was treated with bisoprolol 250 mg/kg/day (High dose group, n=18), and there was also an untreated group (Untreated group, n=18). The survival rate was best in the High dose group. Left ventricular hypertrophy and the expression of proinflammatory cytokines in the myocardium were significantly attenuated in the High dose group, but not in the Low dose group, and oxidative stress was most suppressed in the High dose group. Measurement with electron spin resonance revealed that bisoprolol had a potent scavenging ability, and bisoprolol attenuated the down-regulation of peroxisome proliferation-activated receptor coactivator-1alpha, an important element in the mitochondrial reactive oxygen species detoxification system. CONCLUSION: beta-blocker administration, particularly at high dose, improved the survival rate of the DHF model, at least partly through the attenuation of inflammatory changes and oxidative stress. PMID- 18400559 TI - Transfusion and Apheresis Science. Editorial 38.2. PMID- 18400560 TI - ESFH letter. PMID- 18400558 TI - Prognostic significance of maximum tumour (bulk) diameter in young patients with good-prognosis diffuse large-B-cell lymphoma treated with CHOP-like chemotherapy with or without rituximab: an exploratory analysis of the MabThera International Trial Group (MInT) study. AB - BACKGROUND: The definition and role of bulky disease in young patients (ie, aged 18-60 years) with good-prognosis diffuse large-B-cell lymphoma (DLBCL), who have been treated with CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisolone)-like chemotherapy with or without rituximab, remain controversial. We aimed to assess the effect of maximum tumour diameter (MTD) in these patients. METHODS: Patients from the MInT (Mabthera International Trial Group) study were eligible. We analysed event-free (EFS) and overall survival (OS) after CHOP-like chemotherapy with or without rituximab, according to MTD, by Martingale residual analyses and Cox regression models. Radiotherapy was given to sites of primary bulky disease according to national standards, and to primary extranodal disease at physician discretion. The primary endpoint was EFS and the secondary endpoint was OS. Analyses were by intention to treat. FINDINGS: Of the 824 patients enrolled in the MInT study, the informed-consent form of one patient was missing, leaving 823 patients evaluable for intention-to-treat analysis. Data on MTD of involved sites were available for 802 patients. Martingale residual analysis showed an adverse prognostic effect of MTD on EFS and OS, which increased linearly. In a multivariable analysis with MTD as a linear regression variable, the effect of MTD was significant after CHOP-like treatment alone for EFS (hazard ratio 1.090 [95% CI 1.051-1.130], p < 0.0001) and OS (1.119 [1.057-1.184], p = 0.0001), and after CHOP-like treatment and rituximab for OS (1.089 [1.003-1.183], p = 0.043), but not for EFS (1.044 [0.991-1.099], p=0.103). For CHOP-like treatment alone, 3-year EFS ranged from 78.2% (MTD < 5.0 cm, 95% CI 68.3-85.4) to 41.3% (MTD > or = 10.0 cm, 31.8-50.4). For CHOP-like treatment and rituximab, 3 year EFS ranged from 83.2% (MTD < 5.0 cm, 72.8-89.9) to 72.7% (MTD > or = 10.0 cm, 63.8-79.7). With CHOP-like treatment alone, 3-year OS decreased from 92.9% (MTD < 5.0 cm, 84.9-96.8) to 73.5% (MTD > or = 10.0 cm, 63.9-81.0); for CHOP-like treatment and rituximab, 3-year OS decreased from 98.0% (MTD < 5.0 cm, 92.2-99.5) to 85.2% (MTD > or = 10.0 cm, 77.0-90.6). For CHOP-like treatment, any cut-off point between 5.0 cm and 10.0 cm separated two populations with a significant EFS difference (p < 0.0001 for all log-rank tests) and OS difference (p < or = 0.003 for all log-rank tests). For CHOP-like treatment and rituximab, only a cut-off point of 10.0 cm separated two populations with a significant EFS difference (log rank p = 0.047), but any cut-off point of 6.0 cm or more separated two populations with a significant OS difference (log-rank p values 0.0009-0.037). INTERPRETATION: Rituximab decreased, but did not eliminate the adverse prognostic effect of MTD in young patients with good-prognosis DLBCL. Due to the linear prognostic effect of MTD on outcome, arbitrary cut-off points for bulky disease can be set between 5.0 cm and 10.0 cm, depending on clinical considerations. Based on this study, a cut-off point of 10.0 cm might be a suitable margin in the rituximab era to delineate those patients with bulky disease. PMID- 18400561 TI - Maternal morbidity and mortality in in-vitro fertilization. AB - Pregnancies following in-vitro fertilization (IVF) are known to be at increased risk of a number of pregnancy- and delivery-related complications when compared with non-IVF pregnancies. Most of these complications seem to be due to underlying fertility problems. Ovarian stimulation carries a marked risk for two serious conditions - ovarian torsion and ovarian hyperstimulation syndrome - both of which are relatively rare. Although some common pregnancy complications show an up to five times increased risk over non-IVF pregnancies, the absolute frequencies are still low for most of these conditions. However, an increased risk of placenta praevia might be to some extent due to the IVF procedure. No long-terms effects on cancer risk or mortality can be linked to the IVF procedure, although follow-up time is still relatively short. PMID- 18400562 TI - Effect of bacterial infection on antioxidant activity and lipid peroxidation in the midgut of Galleria mellonella L. larvae (Lepidoptera, Pyralidae). AB - Bacillus thuringiensis is one of the most widely used sources of biorational pesticides, as well as a key source of genes for transgenic expression to provide pest resistance in plants. In this study the effect of Bacillus thuringiensis ssp. galleriae (Bt) infection on the activity of superoxide dismutase (SOD), glutathione S-transferase (GST), catalase (CAT), concentrations of oxidated and reduced thiols (RSSR/RSH) and malondialdehyde (MDA) was tested in the midgut of Galleria mellonella larvae. We found that Bt infection resulted in increased activities of SOD, GST, malondialdehyde and RSSR/RSH ratio the first day after inoculation. However, catalase activity decreased on the first and following days after bacterial infection by Bt. Our results confirm the hypothesis that Bt infection increases the level of oxidative stress in the larval midgut. In light of this study, it seems possible that oxidative damage contributes to cell death in the midgut during bacteriosis. PMID- 18400563 TI - A comprehensive survey of cadherin superfamily gene expression patterns in Ciona intestinalis. AB - We have carried out a comprehensive survey of the spatiotemporal expression of cadherin superfamily genes in the basal chordate Ciona intestinalis, as an example of a genome-wide expression study of a gene family directly regulating cellular processes in morphogenesis. We found 15 definitely expressed cadherin superfamily genes in the Ciona intestinalis genome. Up to the late gastrula stage, all identified delta-protocadherins and the type II classical cadherin, but not other subfamily members, were zygotically expressed. At later stages, however, all cadherin superfamily genes were expressed in the nervous system. These data are useful for understanding the role of these genes in Ciona development and the evolution of chordates. PMID- 18400564 TI - Charge induced closing of Dionaea muscipula Ellis trap. AB - In terms of bioelectrochemistry, Venus flytrap responses can be considered in three stages: stimulus perception, electrical signal transmission, and induction of mechanical and biochemical responses. When an insect touches the trigger hairs, these mechanosensors generate receptor potentials, which induce solitary waves activating the motor cells. We found that the electrical charge injected between a midrib and a lobe closes the Venus flytrap leaf by activating motor cells without mechanical stimulation of trigger hairs. The mean electrical charge required for the closure of the Venus flytrap leaf is 13.6 muC. To close the trap, electrical charge can be submitted as a single charge or applied cumulatively by small portions during a short period of time. Ion channel blocker such as Zn(2+) as well as an uncoupler CCCP, dramatically decreases the speed of the trap closing a few hours after treatment of the soil. This effect is reversible. After soil washing by distilled water, the closing time of Venus flytrap treated by CCCP or ZnCl(2) decreases back from 2-5 s to 0.3 s, but higher electrical charge is needed for trap closure. The mechanism behind closing the upper leaf of Venus flytrap is discussed. PMID- 18400566 TI - Genetic recombination induced by DNA double-strand break in bacteriophage T4: nature of the left/right bias. AB - The experimental system combining double-strand breaks (DSBs), produced site specifically by SegC endonuclease, with the famous advantages of the bacteriophage T4 rII mutant recombination analysis was used here to elucidate the origin of the recombination bias on two sides of the DSB, especially pronounced in gene 39 (topoisomerase II) and gene 59 (41-helicase loader) mutants. Three sources were found to contribute to the bias: (1) the SegC endonuclease may remain bound to the end of the broken DNA and thus protect it from exonuclease degradation; (2) in heteroduplex heterozygotes (HHs), arising as the recombinant products in the left-hand crosses, the transcribed strands are of rII mutant phenotype, so they, in contrast to the right-hand HHs, do not produce plaques on the lawn of the lambda-lysogenic host; and (3) the intrinsic polarity of T4 chromosome, reflected in transcription, may be a cause for discrimination of promoter-proximal and promoter-distal DNA sequences. It is shown that the apparent recombination bias does not imply one-sidedness of the DSB repair but just reflects a different depth of the end processing. It is inferred that the cause, underlying the "intrinsic" bias, might be interference between strand exchange and transcription. Topoisomerase and helicase functions are necessary to turn the process in favor of strand exchange. The idea is substantiated that the double-stranded to single-stranded DNA transition edge (not ss-DNA tip) serves as an actual recombinogenic element. PMID- 18400565 TI - Identification of pathways controlling DNA damage induced mutation in Saccharomyces cerevisiae. AB - Mutation in response to most types of DNA damage is thought to be mediated by the error-prone sub-branch of post-replication repair and the associated translesion synthesis polymerases. To further understand the mutagenic response to DNA damage, we screened a collection of 4848 haploid gene deletion strains of Saccharomyces cerevisiae for decreased damage-induced mutation of the CAN1 gene. Through extensive quantitative validation of the strains identified by the screen, we identified ten genes, which included error-prone post-replication repair genes known to be involved in induced mutation, as well as two additional genes, FYV6 and RNR4. We demonstrate that FYV6 and RNR4 are epistatic with respect to induced mutation, and that they function, at least partially, independently of post-replication repair. This pathway of induced mutation appears to be mediated by an increase in dNTP levels that facilitates lesion bypass by the replicative polymerase Pol delta, and it is as important as error prone post-replication repair in the case of UV- and MMS-induced mutation, but solely responsible for EMS-induced mutation. We show that Rnr4/Pol delta-induced mutation is efficiently inhibited by hydroxyurea, a small molecule inhibitor of ribonucleotide reductase, suggesting that if similar pathways exist in human cells, intervention in some forms of mutation may be possible. PMID- 18400567 TI - Zinc-decorated silica-coated magnetic nanoparticles for protein binding and controlled release. AB - The aim of this study was to be able to reversibly bind histidine-rich proteins to the surface of maghemite magnetic nanoparticles via coordinative bonding using Zn ions as the anchoring points. We showed that in order to adsorb Zn ions on the maghemite, the surface of the latter needs to be modified. As silica is known to strongly adsorb zinc ions, we chose to modify the maghemite nanoparticles with a nanometre-thick silica layer. This layer appeared to be thin enough for the maghemite nanoparticles to preserve their superparamagnetic nature. As a model the histidine-rich protein bovine serum albumin (BSA) was used. The release of the BSA bound to Zn-decorated silica-coated maghemite nanoparticles was analysed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We demonstrated that the bonding of the BSA to such modified magnetic nanoparticles is highly reversible and can be controlled by an appropriate change of the external conditions, such as a pH decrease or the presence/supply of other chelating compounds. PMID- 18400568 TI - Every gastroenterologist deserves a gastrointestinal pathologist. AB - This review evaluates the current evidence supporting the usefulness of gastrointestinal pathology specialists. Using the discovery of Helicobacter pylori as an example, we show that as new conditions are discovered and old ones are revisited and reclassified, an intimate alliance between clinicians, basic scientists, and pathologists is indispensable. Whilst specialised gastrointestinal pathologists (defined as someone who has received at least a year of training in a section of a pathology laboratory where only tissue from the alimentary tract and liver were examined, who has spent time in a clinical gastroenterology unit, and who has restricted his or her practice to such specimens) are usually well-represented in academic settings, their contribution is becoming increasingly recognised also in private practice. We contend that a more sophisticated or detailed diagnosis actually translate to better care, and provide numerous examples that show not only a clinical benefit to the patient and the gastroenterologist, but also a financial advantage for the payors (patients, insurances). PMID- 18400569 TI - Reprocessing in digestive endoscopy units in Lombardy: results of a regional survey. AB - BACKGROUND AND AIM: Inadequate disinfection of endoscopes and associated instrumentation can result in transmission of infections to patients. The aim of this study is to assess cleaning and disinfection practice in gastrointestinal endoscopy units in public and private institutions in the Lombardy region. METHODS: We drafted a questionnaire which we e-mailed to all gastrointestinal centers in Lombardy. RESULTS: A total of 79 centers responded (77%). All endoscopy units perform manual cleaning before disinfection. Automated endoscope reprocessors are available in 84.4% of the centers. Glutaraldehyde-based disinfectants are the most common employed (67% of centers). The time of exposure to the disinfectant is of 10 min in only 9.8% of the centers. The majority of endoscopy units modify their disinfection procedures in the case of infectious disease patients. In 59% of centers disposable material is only used once. In 37% of the endoscopy units instruments are dried with propanol. CONCLUSIONS: The data collected suggest that cleaning and disinfection practices have improved in recent years, and that there is a good compliance with standard guidelines. There is still room for improvement in equipment, disinfection protocols, and traceability of instruments in order to improve safety for patients and staff. PMID- 18400570 TI - Successful pneumatic dilation of achalasia in pregnancy. PMID- 18400571 TI - Biology of intestinal metaplasia in 2008: more than a simple phenotypic alteration. AB - This review concentrates on one main aspect of cancerization in the oesophagus and stomach: principally, intestinal metaplasia. There are at least two other important pathways that lead to cancer and do not need such a morphological transformation. One is the gastric type of carcinoma on the Lauren classification, which arises directly from the stem cell zone and is the signet ring form of cancer, while the other is spasmolytic polypeptide-expressing metaplasia (SPEM)--spasmolytic polypeptide (TFF2) expressing metaplasia, where the gastric glands become filled with TFF2-expressing cells and may also lead to gastric dysplasia and cancer. The development of intestinal metaplasia is complex. Here, we examine intestinal metaplasia in molecular terms, noting the over-expression of Cdx1, Cdx2, Pdx1, Oct1, TFF3 and the downregulation of Hedgehog signalling; Runx3 is deactivated by epigenetic silencing, and pathways such as Wnt and MARK/ERK are involved. These changes start to explain the principles of the development of intestinal metaplasia and suggest that the regulation of these genes is of importance in the development of gastric cancer. PMID- 18400572 TI - Growth of a bonelike apatite on chitosan microparticles after a calcium silicate treatment. AB - Bioactive chitosan microparticles can be prepared successfully by treating them with a calcium silicate solution and then subsequently soaking them in simulated body fluid (SBF). Such a combination enables the development of bioactive microparticles that can be used for several applications in the medical field, including injectable biomaterial systems and tissue engineering carrier systems. Chitosan microparticles, 0.6microm in average size, were soaked either for 12h in fresh calcium silicate solution (condition I) or for 1h in calcium silicate solution that had been aged for 24h before use (condition II). Afterwards, they were dried in air at 60 degrees C for 24h. The samples were then soaked in SBF for 1, 3 and 7 days. After the condition I calcium silicate treatment and the subsequent soaking in SBF, the microparticles formed a dense apatite layer after only 7 days of immersion, which is believed to be due to the formation of silanol (Si-OH) groups effective for apatite formation. For condition II, the microparticles successfully formed an apatite layer on their surfaces in SBF within only 1 day of immersion. PMID- 18400574 TI - Fetal thrombocytopenia. AB - Fetal thrombocytopenia is most often caused by maternal alloantibodies against fetal platelets crossing the placenta and resulting in platelet destruction. This condition, known as fetal and neonatal alloimmune thrombocytopenia, is usually detected after the birth of a symptomatic child who shows signs of bleeding in the skin or in the brain. In the most severe cases, intracranial hemorrhage leads to severe handicap or death. The challenge for the clinician is to provide preventive treatment in the next pregnancy. The current cornerstone of this treatment is maternal intravenous administration of immunoglobulins during the second half of pregnancy. PMID- 18400575 TI - The use of a nasal rasp to shape a nasal implant. PMID- 18400573 TI - Measurement of fracture callus material properties via nanoindentation. AB - In bone fracture healing, the extent to which the injured bone regains stability and strength depends on the mechanical properties of the tissues that are formed during healing. While many techniques have been used to quantify the overall mechanical behavior of fracture calluses, few data exist on the material properties of individual callus tissues. The overall goal of this study was to quantify these material properties. Nanoindentation was performed at multiple locations across thin (200mum), longitudinal sections of rat fracture callus at 35 days post fracture. Following indentation, sections were stained with alizarin red S and alcian blue to obtain semi-quantitative estimates of tissue mineral content and proteoglycan content, respectively. Indentation moduli varied over three orders of magnitude (0.61-1010MPa) throughout the callus. Much of this variation was due to the presence of multiple tissue types: the indentation moduli of granulation tissue, chondroid tissue and woven bone ranged 0.61-1.27MPa (median=0.99MPa), 1.39-4.42MPa (median=2.89MPa) and 26.92-1010.00MPa (median=132.00MPa), respectively. In regions of alizarin red staining, the indentation modulus was correlated (r=0.62, P=0.04) with stain intensity, suggesting a positive correlation between modulus and mineral content in woven bone. In addition, the indentation modulus of woven bone along the periosteal aspect of the cortex increased with distance from the fracture gap (P=0.004). These results demonstrate the usefulness of nanoindentation in characterizing the elastic properties of the heterogeneous mixture of tissues present in bone fracture callus. PMID- 18400576 TI - Syndromes, disorders and maternal risk factors associated with neural tube defects (I). AB - Fetuses with neural tube defects (NTDs) may be associated with syndromes, disorders, and maternal risk factors. This article provides a comprehensive review of syndromes, disorders, and maternal risk factors associated with NTDs, such as acrocallosal syndrome, autosomal dominant brachydactyly-clinodactyly syndrome, Manouvrier syndrome, short rib-polydactyly syndrome, Disorganization ( Ds )-like human malformations, isolated hemihyperplasia, X-linked NTDs, meroanencephaly, schisis association, diprosopus, fetal valproate syndrome, DiGeorge syndrome/velocardiofacial syndrome, Waardenburg syndrome, folic acid antagonists, diabetes mellitus, and obesity. NTDs associated with syndromes, disorders, and maternal risk factors are a rare but important cause of NTDs. The recurrence risk and the preventive effect of maternal folic acid intake in NTDs associated with syndromes, disorders, and maternal risk factors may be different from those of non-syndromic multifactorial NTDs. Perinatal identification of NTDs should alert one to the syndromes, disorders, and maternal risk factors associated with NTDs, and prompt a thorough etiologic investigation and genetic counseling. PMID- 18400577 TI - Syndromes, disorders and maternal risk factors associated with neural tube defects (II). AB - Fetuses with neural tube defects (NTDs) may be associated with syndromes, disorders, and maternal risk factors. This article provides a comprehensive review of syndromes, disorders, and maternal risk factors associated with NTDs, such as Currarino syndrome, sacral defect with anterior meningocele, Jarcho-Levin syndrome (spondylocostal dysostosis), lateral meningocele syndrome, neurofibromatosis type I, Marfan syndrome, and hyperthermia. The recurrence risk and the preventive effect of maternal folic acid intake in NTDs associated with syndromes, disorders, and maternal risk factors may be different from those of non-syndromic multifactorial NTDs. Perinatal identification of NTDs should alert one to the syndromes, disorders, and maternal risk factors associated with NTDs, and prompt a thorough etiologic investigation and genetic counseling. PMID- 18400578 TI - Medical treatment for uterine myomas. AB - Uterine myomas are the most common benign tumors in the female reproductive tract. Most women with myomas are asymptomatic. Therefore, expectant observation and follow-up are often recommended for these myoma patients. However, myomas may cause menstrual symptoms, pelvic pain, pressure complaints, subfertility or pregnancy-related complications, with resultant requests for a definitive treatment. The management of myomas has become multidisciplinary in the past 20 years. Basically, the choice of treatment depends on the patient's age, the reason for treatment, the issue of fertility preservation, and the patient's preference. The treatment spectrum includes an expectant management, medical therapy, surgical intervention, uterine artery embolization or ablative techniques. Medical therapy is an option for women with symptomatic myomas who prefer non-surgical treatment, consider fertility preservation, or expect a less aggressive operation after shrinkage of the uterine volume. This review will summarize the recent well-documented drugs for the management of uterine myomas. PMID- 18400579 TI - The role of selective estrogen receptor modulators on breast cancer: from tamoxifen to raloxifene. AB - The link between hormones and breast cancer growth and development has been recognized for more than a century. Estrogen stimulates the proliferation of breast epithelial cells, and both endogenous and exogenous estrogens have been implicated in the pathogenesis of breast cancer. Classically, estrogen action at target sites around the body is mediated through related but distinct estrogen receptors (ERs), designated ERalpha and ERbeta, to alter gene expression. This accumulating understanding of the mechanism of action of estrogen led ultimately to the design of antiestrogenic agents that work by virtue of their interaction with the ER; these drugs have come to be known as selective estrogen receptor modulators (SERMs). Tamoxifen, a SERM, emerged as the first antiestrogenic agent that is clinically applicable to breast cancer. Tamoxifen became the "gold standard" and established the principles of tumor targeting and identified the appropriate treatment strategy to aid survivorship in breast cancer patients, with enhancement of disease-free survival and a 50% decrease in recurrences observed in ER-positive patients 15 years after diagnosis. However, because of the many adverse events in the use of tamoxifen, some of which have contributed to significant morbidity and mortality, drug modification which has resulted in fewer incidences of adverse events without compromising the therapeutic effect for breast cancer prevention may face an easier road to acceptance. Raloxifene may be a better alternative, since evidence from large clinical trials showed that raloxifene not only decreases the incidence of osteoporosis and related fractures, but also offers benefits for breast cancer prevention. The results from the Study of Tamoxifen and Raloxifene (STAR) trial showed the superiority of raloxifene over tamoxifen, not only for the equal efficacy in the prevention of invasive breast cancer but also for the fewer serious adverse events. Taken together, without other competition so far, raloxifene is recommended for postmenopausal women with osteoporosis who also need breast cancer prevention. PMID- 18400580 TI - Comparison of whole genome amplification methods for further quantitative analysis with microarray-based comparative genomic hybridization. AB - OBJECTIVE: Whole genome amplification (WGA) is a crucial procedure for genomic DNA analysis from limited sources, such as in forensic analysis, embryo biopsy for preimplantation genetic diagnosis, or needle aspiration biopsies. Several strategies for WGA have been developed for either genotyping or microarray-based comparative genome hybridization (array-CGH) during the past decade. Nevertheless, there were few studies in which various WGA methods had been performed side-by-side and results evaluated with multiple methods. MATERIALS AND METHODS: Ease of performance, qualitative accuracy, and quantitative fidelity of different WGA methods, such as degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), ligation-mediated PCR (LM-PCR) and strand displacement amplification (SDA), were compared in amplifying genomic DNA derived from karyotype-confirmed amniocytes and the cancer cell line SAOS2. RESULTS: Using analysis with microsatellite markers, single nucleotide polymorphism markers, and array-CGH, our results suggested that: (1) genomic DNA amplified from DOP-PCR resulted in false positive and negative results by analysis with array-CGH; (2) SDA is the easiest performance method among the three WGA methods; and (3) amplified DNA products generated by LM-PCR best reflect the original genomic DNA. CONCLUSION: The amplified DNA products generated by LM-PCR best reflect the original genomic DNA. PMID- 18400581 TI - Abdominal wall endometriosis: an overlooked but possibly preventable complication. AB - OBJECTIVE: To find ways of preventing abdominal wall endometriosis through a retrospective case review. MATERIALS AND METHODS: A retrospective study of 22 patients presenting with 26 postoperative abdominal wall masses. All masses were pathologically proved to be scar endometriosis between September 1994 and September 2006. The age, parity, symptoms and duration, previous surgeries, interval between previous surgery and current operation, initial diagnosis, and the 26 sites and size of endometrioma were recorded and analyzed. RESULTS: About 60% of the patients were in the fourth decade of life. All 22 cases, except one with mid-trimester hysterotomy, had previous cesarean section (CS). Three cases had vertical midline incision for CS, and the other 19 had Pfannenstiel incision, 18 of which were for CS and one for hysterotomy. Of the 22 cases, only three had multiple endometriomas, i.e. one case had three foci and the other two cases had two foci each. Twenty-three endometriomas were found in the Pfannenstiel incision group; 19 out of the 23 foci (82%) were located in either corner of the Pfannenstiel incision wounds (with right side predominance in 13 out of 19). Three endometriomas were noted in vertical midline incisions and two were in the upper corner. Three endometrioma excisions were done during repeated CS. CONCLUSION: Abdominal wall endometriosis may be caused by iatrogenic inoculation of the endometrium into the surgical wound. It is strongly recommended that, at the conclusion of the surgical procedure, the abdominal wound be cleaned thoroughly, particularly at both corner sites (especially the operators side). If an abdominal wall endometriosis is encountered after CS but the patient plans to have future pregnancy and the symptoms are mild, excision of the endometrioma may be deferred until the next indicated CS. PMID- 18400582 TI - Reappraisal of fetal abdominal circumference in an Asian population: analysis of 50,131 records. AB - OBJECTIVE: Fetuses from different populations may show different growth patterns. In obstetrics, fetal abdominal circumference (AC) is a very useful index for assessing fetal growth. In this study, we attempted to establish the normal fetal growth curves of AC in an Asian population in South Taiwan. MATERIALS AND METHODS: We reviewed our computer ultrasound database of fetal AC records from January 1991 to December 2006. During the study period of 16 years, only the fetuses examined by ultrasonography with gestational age between 14 and 41 weeks were included. We excluded extreme bilateral records after initial analysis. Eventually, 50,131 records of AC were included for final analysis. RESULTS: The observed gestation-specific AC values and the predicted AC values were calculated. The best-fit regression equation of AC versus gestational age is a second-order polynomial equation. In general, fetal AC values in our population showed similar patterns to those in Western populations. Besides, we established a table of the predicted AC values based on specific gestational age, including the 5 th , 10 th , 50 th , 90 th and 95 th centiles, for clinical reference. CONCLUSION: To the best of our knowledge, our series is the largest sample of AC reported in the medical literature. We believe that the gestational age-specific nomogram of fetal AC is important for further clinical assessment of fetal growth. PMID- 18400583 TI - Comparison of single versus double intrauterine insemination. AB - OBJECTIVE: To compare the outcomes of single versus double intrauterine insemination. MATERIALS AND METHODS: This prospective randomized study was carried out in 100 infertile patients. One intrauterine insemination was applied 36 hours after human chorionic gonadotropin (hCG) injection to 50 patients in the first group. To 50 patients in the second group, two intrauterine inseminations were applied, of which the first was applied 24 hours after and the second 48 hours after the hCG injection. RESULTS: In the first group, pregnancies were detected in eight patients (pregnancy rate per patient was 16%, pregnancy rate per cycle was 10.6%). In the second group, pregnancies were detected in five patients (pregnancy rate per patient was 10%, pregnancy rate per cycle was 6.4%). There was no statistically significant difference between the two groups ( p > 0.05). CONCLUSION: Single intrauterine insemination can be considered to be more reasonable than double intrauterine insemination treatment, taking into consideration the economic cost and the psychologic trauma to the patients. However, further studies with larger sample sizes are needed in order to reveal any actual differences between the two methods. PMID- 18400584 TI - Relationship between uterine and ovarian arterial blood flow measured by Doppler sonography at different stages of puberty. AB - OBJECTIVE: Measuring blood flow using a special form of color Doppler ultrasonography can help radiologists to identify the stages of development of organs, such as the uterus and ovary. The aim of this study was to determine the relationship between arterial blood flow to the uterus and ovaries at different stages of puberty. MATERIALS AND METHODS: Sixty girls referred to the Children's Medical Center in Tehran for the evaluation of pubertal stages were included in the study. They were divided into three subgroups: girls without primary puberty signs ( n = 20); girls with these puberty signs but without the beginning of menstruation ( n = 20); and girls with signs of puberty and menstruation ( n = 20). Uterus and ovary volumes were measured with the use of a 3.5-MHz convex transducer (Logic 500 Pro; General Electric, Milwaukee, WI, USA) and Doppler flow measurements of the uterine vessels were performed transabdominally with a 3.5 MHz color Doppler system (AU4 Idea; Esaote Biomedica, Genoa, Italy). RESULTS: We found no significant differences between the groups in terms of either resistance index or pulsatility index of the right ovary, left ovary or uterus. However, there were relationships between the volume of the uterus and pubertal stages, and also between the volume of the ovaries and these stages. CONCLUSION: Uterine and ovarian arterial blood flow, measured by Doppler sonography, was not useful for the evaluation of pubertal stages. However, it should be investigated with a greater sample size. PMID- 18400585 TI - Comparisons of different dosages of gonadotropin-releasing hormone (GnRH) antagonist, short-acting form and single, half-dose, long-acting form of GnRH agonist during controlled ovarian hyperstimulation and in vitro fertilization. AB - OBJECTIVE: Both gonadotropin-releasing hormone (GnRH) analogs and antagonists have been used for pituitary desensitization during controlled ovarian hyperstimulation (COH). We aimed to determine the minimum effective daily dose of GnRH antagonist in women undergoing COH. We also compared the efficiency of a GnRH antagonist and a GnRH agonist. MATERIALS AND METHODS: Women undergoing in vitro fertilization/intracytoplasmic sperm injection and embryo transfer were divided into five groups: (1) cetrorelix 0.25 mg ( n = 86); (2) cetrorelix 0.2 mg ( n = 28); (3) cetrorelix 0.15 mg ( n = 30); (4) leuprolide acetate (LA) 0.5 mg/day ( n = 58); (5) single half-dose LA depot 1.88 mg ( n = 49). Cetrorelix was administered daily from menstrual day 8 until the day of human chorionic gonadotropin administration. LA or LA depot was started on day 21 of the previous menstrual cycle. RESULTS: We observed lower gonadotropin (Gn) dosages, estradiol (E2) levels and reduced risk of ovarian hyperstimulation syndrome (OHSS) in the GnRH antagonist groups. A higher risk of luteinizing hormone (LH) surge was noted in cetrorelix 0.2 and 0.15 mg groups. Gn dosages (IU)/E2 levels (pg/mL) in each group were: (1) 1,949.4/1,191.1; (2) 1,869.6/1,010.8; (3) 1,856.7/1,023.6; (4) 2,184.5/1,323.6; and (5) 2,103.5/1,313.5, respectively. LH/OHSS risks were: (1) 3.5%/5.8%; (2) 7.1%/3.6%; (3) 13.3%/3.3%; (4) 3.4%/8.6%; and (5) 2%/8.2%, respectively. Number of oocytes/embryos/grade I, II embryos were: (1) 9.4/7.9/5.8; (2) 7.5/4.2/3.6; (3) 6.3/4.1/3.1; (4) 12.3/8.9/6.6; and (5) 11.8/8.4/6.1, respectively. There was no significant difference in terms of clinical outcomes between groups 1, 4 and 5, except for higher abortion rates (AR) in group 1. Pregnancy rate (PR)/implantation rate (IR) ratios in groups 1, 4, and 5 were statistically higher than those in groups 2 and 3. Chemical PR/IR/AR were: (1) 30.2%/5.9%/7%; (2) 21.4%/5.1%/7.1%; (3) 16.7%/4.1%/10%; (4) 32.8%/5.5%/8.6%; and (5) 30.6%/5.7%/8.2%, respectively. CONCLUSION: The lowest effective dosage of cetrorelix for pituitary desensitization during COH luteolysis is 0.25 mg, resulting in a comparable PR but a higher AR when compared with GnRH agonist. PMID- 18400586 TI - The application of three-dimensional ultrasonography in the prenatal diagnosis of arthrogryposis. AB - OBJECTIVE: To present the application of three-dimensional (3D) ultrasonography in the early prenatal diagnosis of fetal arthrogryposis. CASE REPORT: A 26-year old multipara had a fetus with anomalies of the limbs as shown by conventional ultrasonography at 18 weeks of gestation. A follow-up 3D ultrasonogram at the same gestational age was consistent with the diagnosis of arthrogryposis. Based on an abnormal chromosome 18p on the fetal karyotype, termination of the pregnancy was performed at 22 weeks' gestation. The outward appearance of the fetus coincided with the prenatal sonographic findings of arthrogryposis. CONCLUSION: As an advanced and sophisticated technology, 3D ultrasonography can serve as a useful technique for the early diagnosis of fetal anomalies, including arthrogryposis. The earlier the diagnosis is established, the earlier the appropriate management can be initiated, including counseling, additional work-up and timely termination of pregnancy, if indicated. PMID- 18400587 TI - Late prenatal diagnosis of hydrometrocolpos secondary to a cloacal anomaly by abdominal ultrasonography with complementary magnetic resonance imaging. AB - OBJECTIVE: Prenatal diagnosis of a cloacal anomaly is difficult. Magnetic resonance imaging (MRI) can assist in the identification of the connection and continuity of a cystic mass to confirm the diagnosis of a cloacal anomaly. CASE REPORTS: In the first case, a fetal abdominal cystic mass was observed at 32 weeks of gestation. Ultrasonography revealed a retrovesical septate hypoechoic mass with bilateral hydronephrosis. MRI demonstrated a midline cystic mass connected to a dilated uterus and a possible fistula between the bladder and vagina. In the second case, a fetal abdominal septate cystic mass was identified using ultrasonography at 34 weeks of gestation. MRI was performed and demonstrated hydrocolpos/hydrometrocolpos originating from a uterine didelphis with left dysgenesis and a possible vesicovaginal fistula. After birth, both newborns underwent immediate surgical intervention with good outcomes. CONCLUSION: MRI facilitated the prenatal diagnosis of cloacal anomalies and allowed additional time for parental counseling and planning of the delivery method with subsequent neonatal intensive care and surgical and urologic consultations. PMID- 18400588 TI - A rare case of rudimentary uterus with absence of both ovaries and 46,XX normal karyotype without mosaicism. AB - OBJECTIVE: We report an 18-year-old patient with bilateral ovarian agenesis, rudimentary uterus and normal fallopian tubes, and with normal 46,XX karyotype (without mosaicism). CASE REPORT: The patient was admitted to our clinic with primary amenorrhea. Secondary sexual characteristics (thelarche and pubarche) were both Tanner classification stage 1. With the help of vaginoscopy, vaginal depth was measured without distorting the hymenal ring and was found to be 8 cm. The laboratory findings were as follows: follicle-stimulating hormone 85 IU/L, luteinizing hormone 40 IU/L, and estradiol 14 pg/dL. Genetic investigation revealed a 46,XX karyotype without any mosaicism. Diagnostic laparoscopy was performed. During laparoscopic pelvic exploration, a rudimentary uterus without ovaries and normal bilateral fallopian tubes were observed. Bone mineral densitometry measurements were in the normal range. The patient was given oral contraceptives for hormone replacement. CONCLUSION: If gonadal agenesis is thought to be the cause of primary amenorrhea in patients with defective secondary sexual characteristics, we believe that laparoscopic evaluation is the gold standard in diagnosis. PMID- 18400589 TI - Intrauterine fetal goiter: diagnosis and management. AB - OBJECTIVE: The diagnosis and management of a fetal neck mass encountered during routine antenatal ultrasound screening, and later diagnosed definitely as a intrauterine fetal goiter, is discussed in this case report. CASE REPORT: A 28 year-old primigravida at 36 weeks' gestation with normal vital findings, normal laboratory values, and no history of pathologic findings was examined in our outpatient department for the first time. Her previous antenatal visits were done in secondary maternal care units. The ultrasound screening revealed polyhydramnios (amniotic fluid index, 21 cm), a homogeneous solid mass of 3.67 x 7.88 cm on the anterior of the fetal neck, and fetal abdominal ascites. Other fetal organs appeared normal. Maternal thyroid panel was in the normal range. The fetus was delivered abdominally at 38 weeks' gestation and diagnosed with defective thyroid hormone biosynthesis. With appropriate treatment, the size of the thyroid gland was observed to decrease. CONCLUSION: The early diagnosis and appropriate treatment of intrauterine fetal goiter is crucial, owing to the potential mental and motor outcomes of this disease. PMID- 18400590 TI - Prenatal sonographic and magnetic resonance imaging demonstration of a right midline arachnoid cyst associated with ventriculomegaly, colpocephaly, dilation of the third ventricle, absence of cavum septi pellucidi, agenesis of the corpus callosum and mega cisterna magna. PMID- 18400591 TI - Concomitant omphalocele and anencephaly associated with trisomy 18 and arthrogryposis diagnosed in early pregnancy. PMID- 18400592 TI - Prenatal visualization of cebocephaly with a prominent nose in a second-trimester fetus with alobar holoprosencephaly and trisomy 13. PMID- 18400593 TI - Posterior reversible encephalopathy syndrome in a pregnant woman. PMID- 18400594 TI - Noncardiogenic pulmonary edema due to ritodrine usage in preterm labor. PMID- 18400595 TI - Delayed recurrence of cervical cancer after radical hysterectomy for carcinoma in situ. PMID- 18400596 TI - Multicystic encephalomalacia in an uncomplicated twin pregnancy. PMID- 18400597 TI - Transarterial embolization for the control of postpartum hemorrhage. PMID- 18400598 TI - Successful clinical application of the molecular adsorbent recirculating system in a patient with acute fatty liver of pregnancy. PMID- 18400599 TI - Obstructive uropathy as a major presentation of ovarian clear cell adenocarcinoma. PMID- 18400600 TI - Postablation tubal sterilization syndrome. PMID- 18400601 TI - Disseminated peritoneal leiomyomatosis. PMID- 18400602 TI - Prenatal diagnosis and management of twin reversed arterial perfusion (TRAP) syndrome. PMID- 18400603 TI - Tamoxifen: to use or not to use. PMID- 18400604 TI - Is neonatal Bacillus Calmette-Guerin vaccination protective in Taiwan? PMID- 18400605 TI - Molecular epidemiology of human papillomavirus. AB - Human papillomavirus (HPV) has been considered to be an etiologic factor for anogenital cancers, such as cervical cancer and possibly a subset of cancers of the aerodigestive tract. These small, non-enveloped, double-stranded DNA viruses primarily infect the epithelium and induce benign as well as malignant lesions of the mucosa and skin. Some HPVs are considered to be high-risk due to their strong implication in carcinogenesis, particularly the malignant progression of cervical tumors. The recognition of papillomaviruses as a major etiologic agent for human cancers has increased their medical importance and stimulated research into developing strategies for the screening, diagnosis, prevention and treatment of HPV-associated diseases. PMID- 18400606 TI - Improvement in timing of antibiotic administration by using a prophylactic antibiotic record form. AB - BACKGROUND/PURPOSE: To evaluate the effect of a form recording the timing of prophylactic antibiotic administration, antibiotics used, and treatment duration on the proper use of prophylactic antibiotics in patients undergoing four types of operations. METHODS: This was a retrospective study in a 682-bed, university affiliated, tertiary teaching hospital. Using the form developed, nurse practitioners recorded the timing of prophylactic antibiotic administration, antibiotics used, and treatment duration for patients undergoing three types of clean operations (total hip replacement, total knee replacement, thyroidectomy) and one type of clean-contaminated operation (hemorrhoidectomy). Patient records were identified by chart review after searching the hospital database. A case was defined by a principal or secondary procedure code for the four types of operations. Patients who received therapeutic antibiotics for documented preoperative infection were excluded. RESULTS: A significant improvement (p<0.001; OR, 33.768; 95% CI, 4.304-264.951) in the use of prophylactic antibiotics was noted between the pre-intervention and intervention periods. Among the three criteria of antibiotic usage, the improvement in the appropriate timing of prophylactic antibiotics was significant (p<0.001; OR, 46.247; 95% CI, 5.891-363). The accuracy of the prophylactic antibiotic form in recording the timing of antibiotic administration, the choice of antibiotics, and the treatment duration was 84.2% (16/19), 89.4% (17/19), and 100% (19/19), respectively. CONCLUSION: The prophylactic antibiotic form effectively decreased the inappropriate use of prophylactic antibiotics. Due to its high degree of reliability, this infection control measure can be used as a continuous monitoring system for prophylactic antibiotic use. PMID- 18400607 TI - Clinical features and predictors of a complicated treatment course in peripheral tuberculous lymphadenitis. AB - BACKGROUND/PURPOSE: There remains uncertainty regarding the treatment strategy for patients with peripheral tuberculous lymphadenitis (pTBL) in areas endemic for tuberculosis. The purpose of this study was to demonstrate the clinical features and assess the predictors of a complicated treatment course in pTBL. METHODS: A retrospective analysis of 97 pTBL patients from January 1995 through to December 2004 was conducted. Patient characteristics with and without a complicated treatment course, defined as prolonged treatment (>9 months) and/or relapse, were compared for determining the predictors. RESULTS: The disease occurred predominantly in females (57.7%) with a mean age of 37. Most patients (72%) were asymptomatic. Cervical nodes were the most common (72%) manifestations. Fifty-six patients completed a 6-9 month course of therapy without relapse; 28 had a prolonged but complete treatment course, and 13 relapsed within a mean of 8.5 months after treatment (range, 3-42 months; median, 7.8 months). Of 97 pTBL patients, six had enlarged or newly appeared lymph nodes during treatment. Multivariate analysis indicated that low body mass index and bilateral cervical nodes were independent determinants of a complicated treatment course with the odds ratios of 1.2 (95% CI, 1.01-1.41; p=0.042) and 3.9 (95% CI, 1.08-14.0; p=0.038), respectively. CONCLUSION: This study found that pTBL is more likely to occur in young female patients. For patients who present with bilateral cervical nodes and low body mass index, a prolonged treatment course to ensure disease control should be considered. PMID- 18400608 TI - Psychometric properties of the Chinese version of the Perceived Therapeutic Efficacy Scale for type 2 diabetes. AB - BACKGROUND/PURPOSE: The purpose of this study was to test the psychometric properties of the Perceived Therapeutic Efficacy Scale (PTES) for type 2 diabetes with a Taiwanese sample. The mortality rate and health care cost of diabetes have dramatically increased in Taiwan, with many people with diabetes lacking the ability to control their disease appropriately. Addressing this problem requires enhancing self-efficacy towards self-management. Thus, there is a particular need for research into developing a diabetes-specific self-efficacy measurement instrument in Taiwan. METHODS: This study was undertaken in two stages. Stage 1 consisted of forward and back translation of the PTES into Chinese and examination of content validity. Stage 2 established the validity and reliability of the Chinese version of PTES (C-PTES). A total of 230 people with type 2 diabetes aged 30 years or more from a diabetes outpatient clinic and taking oral medicine were recruited for psychometric testing. RESULTS: Significant criterion related validity was demonstrated between the C-PTES and the Summary of Diabetes Self-Care Activities scores (r=0.32; p<0.01). Convergent validity was confirmed as the C-PTES converged well with the General Self-Efficacy Scale in measuring self-efficacy (r=0.42; p<0.01); construct validity using factor analysis composed a single subscale. Internal consistency showed Cronbach's alpha was 0.95 and the test-retest reliability (Pearson's correction) was 0.79 (p<0.01) and a Bland Altman plot showed that 97% of the subjects were within two standard deviations of the mean. CONCLUSION: The results of reliability and validity strengthen confidence in using the C-PTES. The C-PTES requires future studies to confirm the psychometric properties. PMID- 18400609 TI - Detection of polyomavirus JC genotype from transplant patients by capillary electrophoresis: comparison to fragment length polymorphism analysis. AB - BACKGROUND/PURPOSE: Among the genotypes of human polyomavirus JC (JCV) reported in Taiwan, CY, TW1, TW2 and TW3 are the most commonly correlated with human diseases. JCV is usually detected using nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis. The aim of this study was to detect the rate of positivity and genotype of the JCV genome in urine by RFLP or capillary electrophoresis (CE) in renal transplant patients and healthy volunteers. METHODS: We compared CE analysis to the methods of nucleotide sequencing and RFLP analysis for detection of JCV viruria among 60 renal transplant patients and 50 unrelated healthy controls. Genotyping of the positive PCR products was performed using CE and RFLP analysis simultaneously. RESULTS: The urine JCV-positive rate was significantly higher in renal transplant patients than in healthy volunteers (40% [24/60] vs. 20% [10/50]; p=0.0238). In addition, multiple genotypes of JCV could be detected by CE, but only one genotype could be detected by RFLP. In our study, 20% (2/10) of urine JCV-positive samples from healthy volunteers had two different genotypes. In renal transplant patients 66% (16/24) of JCV-positive samples had two different genotypes and 12% (3/24) had three different genotypes. CONCLUSION: In comparison with RFLP, CE can detect multiple genotypes in urine JCV-positive samples and requires only 1/200 of the volume of specimen required for RFLP analysis. The CE method has sensitivity and specificity suitable for use in the clinical laboratory, and identifies more genotypes than RFLP analysis. PMID- 18400610 TI - MMP-9 mRNA as a therapeutic marker in acute and chronic stages of arthritis induced by type II collagen antibody. AB - BACKGROUND/PURPOSE: Antibodies against type II collagen (anti-CII) are arthritogenic and central to the initiation of the disease. An animal model of collagen type II-specific monoclonal antibody-induced arthritis (CAIA) has been used for the evaluation of various therapeutic effects in rheumatoid arthritis (RA). We aimed to measure the expression of matrix metalloproteinase (MMP)-9 (gelatinase B), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL 1beta) in the acute and chronic stages of the CAIA model for application as a therapeutic marker. METHODS: A commercially available antibody cocktail containing four monoclonal anti-type II collagen antibodies were injected into 6 to 8-week-old male BALB/c mice (n=20) and 50 microL lipopolysaccharide was injected 3 days later. The clinical manifestations of RA were recorded and scored at 10 days (acute stage) and 21 days (chronic stage). Then the mice were sacrificed for histologic analysis of the inflamed footpad and gene expression of IL-1beta, TNF-alpha and MMP-9 by ELISA and quantitative polymerase chain reaction amplification. RESULTS: Marked inflammation was found in the limb joints of mice at 10 days. Both IL-1beta and MMP-9 expression played a central role in the inflammatory reaction in the acute stage. The expression level of MMP-9 mRNA remained high in the chronic stage of CAIA, but that of IL-1beta mRNA was unexpectedly negligible; the serum level of TNF-alpha in CAIA was undetectable in the acute stage. The expression level of TNF-alpha mRNA was also lower than IL 1beta and MMP-9 in the acute inflammatory stage. CONCLUSION: The CAIA model is a fast and highly replicable model of RA. MMP-9 and IL-1beta were highly expressed in the acute stage of CAIA. It is suggested the MMP-9 mRNA level is a suitable marker for both acute and chronic stage, whereas IL-1beta is a marker only for the acute stage of the CAIA murine model. PMID- 18400611 TI - Safety and feasibility of drug-eluting stent implantation at vertebral artery origin: the first case series in Asians. AB - BACKGROUND/PURPOSE: Studies of the safety and efficacy of drug-eluting stent (DES) implantation to treat vertebral artery (VA) ostial stenosis are lacking in the Asian population. The aim of this study was to evaluate the feasibility and safety of DES implantation to treat VA ostial stenosis in Asians. METHODS: We retrospectively analyzed the medical and radiologic records of patients receiving VA origin stenting with DES from October 2004 to September 2006 in the National Taiwan University Hospital. RESULTS: Eleven symptomatic patients (9 male; mean age, 72 years) with 11 lesions treated with DES were included. There were eight lesions treated with paclitaxel-eluting stents and three lesions treated with sirolimus-eluting stents. The success rate for implantation of a DES was 100%. The percentage of stenosis was significantly reduced after stenting (86+/-5 to 2+/-4%; p<0.001). No peri-interventional neurologic, vascular, or other medical complications were noted. All patients were asymptomatic at the mean follow-up time of 18.7+/-8.6 months (range, 6-30 months). Two patients received repeat angiography at 4 or 8 months after stenting. No significant in-stent restenosis was found. CONCLUSION: Symptomatic VA origin stenosis can be treated safely and effectively with DES in Asians. PMID- 18400612 TI - Increasing drug resistance of Mycobacterium tuberculosis isolates in a medical center in northern Taiwan. AB - BACKGROUND/PURPOSE: This retrospective study was conducted to evaluate the drug resistance patterns of Mycobacterium tuberculosis in a medical center in northern Taiwan between 2003 and 2004 in comparison to those reported in 1990-1992. METHODS: A total of 611 non-duplicate M. tuberculosis isolates from culture proven tuberculosis (TB) cases were tested for drug susceptibility against five first-line anti-TB drugs in a clinical mycobacterial laboratory using the agar proportional method for isoniazid (INH), rifampicin (RIF), ethambutol (EMB), and streptomycin (SM). The Wayne assay, which measures the activity of pyrazinamide (PZA), was used for PZA susceptibility testing. RESULTS: Of 611 patients, including 510 males and 101 females, 70.2% of patients were older than 65 years. A total of 339 isolates (55.5%) were resistant to one or more drugs. Isolates from patients aged <25 years showed a significantly higher drug resistance rate (79.2%) compared with other age groups (p=0.0312). Single-drug resistance was observed in 97 (15.9%) of all isolates. Monoresistance to PZA (8.0%) was most frequent, followed by INH (5.1%), RIF (0.5%), EMB (1.6%), and SM (0.7%). Among the polydrug resistant isolates (PDR-TB), resistance rates were 35.5% for INH and 27.0% for RIF. One hundred and fifty-nine isolates (26.0%) were resistant to both INH and RIF (multidrug-resistant [MDR] TB); 94.6% of RIF-resistant isolates were also resistant to INH. The overall drug resistance rates and percentages of PDR TB and MDR-TB increased over the 12-year study period (p<0.001). Based on medical records, primary cases were identified in 486 (84.7%) out of 574 patients, and resistance to any drug was identified in 268 (55.1%) patients, of which 130 (26.7%) were MDR-TB. Among the 88 with recurrent TB, 54 (61.4%) were resistant to at least one drug, and MDR-TB was identified in 29 (33.0%) patients. A history of previous anti-TB therapy was a significant factor for overall drug resistance, PZA monoresistance, PDR-TB, and MDR-TB (p<0.001). CONCLUSION: The emergence of M. tuberculosis isolates resistant to anti-TB agents in this hospital, and in particular among young patients, is alarming. Strict measures to control and prevent drug-resistant TB are urgently needed. PMID- 18400613 TI - Crossed pulmonary arteries: report of two cases with emphasis on three dimensional helical computed tomographic imaging. AB - Crossed pulmonary arteries are due to an anomalous origin of both pulmonary arteries from the main pulmonary trunk. This anatomy is often associated with other congenital cardiac and extracardiac diseases. We report two neonates with complex congenital heart disease who had this disorder, which was detected during cardiac computed tomography (CT) with three-dimensional reconstruction but not during echocardiography or angiography. The first patient was a 3-day-old male neonate who had tachypnea and feeding problems since birth. Cardiac CT showed crossed pulmonary arteries, type B interruption of the aortic arch, a ventricular septal defect, and a large patent ductus arteriosus. He received an emergency T colostomy at 3 days of age because of severe bowel distention. Low-type imperforated anus was diagnosed. His postoperative course was complicated with fluctuated saturation, seizure, hypocalcemia, hyperphosphatemia, and sepsis. Also found were cardiac defects, abnormal facies, thymic hypoplasia, cleft palate, hypocalcemia, and a variable deletion on chromosome 22q11 (CATCH 22 disorder). Because of his poor prognosis, the patient was transferred to another hospital on day 16 for further care, at the family's request. The other patient was a 5-day old female neonate who had a heart murmur since birth. Cardiac CT showed crossed pulmonary arteries, truncus arteriosus, type A interruption of the aortic arch, a ventricular septal defect, an atrial septal defect, and a large patent ductus arteriosus. She received complete surgical correction, including division of the patent ductus arteriosus and repair of the other defects. Intermittent respiratory distress and decreased blood pressure complicated her postoperative course, and she died on the eighth day after surgery. Crossed pulmonary arteries complicated accurate interpretation of two-dimensional echocardiographs of the great vessels, as well as the course and location of catheters during cardiac catheterization. Three-dimensional CT provided a noninvasive approach to clearly recognize these malformations and the related anatomic structures. This information is important in planning and performing surgery in neonates with crossed pulmonary arteries. PMID- 18400614 TI - Nephrogenic systemic fibrosis associated with gadolinium use. AB - Nephrogenic systemic fibrosis (NSF) is an idiopathic, progressive, systemic fibrosis that occurs in patients with renal diseases. Recently, gadolinium containing contrast (Gd-contrast) has become a suspected causal factor for NFS. This report discusses two female patients with end-stage renal disease, aged 70 and 51 years, respectively, who developed histologically proven NSF after exposure to Gd-contrast. Clinically, both patients were characterized by fibrosis and induration of skin and muscle mainly in the limbs with joint contracture. In the first case, NSF developed gradually after undergoing evaluation by Gd contrast magnetic resonance imaging (MRI) and subsequent surgery for her urothelial carcinoma. In the second patient, NSF developed after undergoing evaluation by Gd-contrast MRI for her right shoulder bursitis with calcification, and the conditions of NSF continued to worsen after the surgical treatment of this right shoulder lesion. Although the role of Gd-contrast in NSF is still not well known, the correlation in our cases strongly suggests that it should be used with cautioned in patients with end-stage renal disease. Both of our patients underwent surgery before or during the development of NSF, indicating that the surgical procedure may be a contributing factor. PMID- 18400615 TI - Notice of retraction of "Significant association of HLA-DQ5 with autoimmune hepatitis in Taiwan", J Formos Med Assoc 2007;106(12):1063-8. PMID- 18400616 TI - [E-page Original Images. An interesting case of coronary artery fistula: diagonal artery-, left circumflex artery- and right coronary artery to left ventricle fistulas]. PMID- 18400617 TI - [E-page Original Images. A case of coronary artery aneurysm resulted in sudden death]. PMID- 18400618 TI - E-page Original Images. Thrombotic lesion of saphenous vein graft resolved by antiaggregant therapy. PMID- 18400619 TI - E-page Original Images. The accordion phenomenon. PMID- 18400620 TI - E-page Original Images. A rare radiodiagnostic finding: totally and severely calcified aorta--the Porcelain aorta. PMID- 18400621 TI - E-page Original Images. The imaging of the aortic fibrous membrane associated with Williams syndrome. PMID- 18400622 TI - E-page Original Images. Left pulmonary artery thrombosis diagnosed by transesophageal echocardiography. PMID- 18400623 TI - E-page Original Images. Percutaneous coronary intervention performed for the revascularization of a single coronary artery originating from the right sinus of Valsalva. PMID- 18400624 TI - E-page Original Images. Coronary artery evaluation in Kawasaki disease by dual source multi-detector CT coronary angiography in children. PMID- 18400625 TI - E-page Original Images. Single coronary artery diagnosed by multidetector computed tomography. PMID- 18400626 TI - [Co-editors participation, preparation of the new editorial board, where is the ethic? Our new affliction]. PMID- 18400627 TI - Citric acid as a decalcifying agent for the excised calcified human heart valves. AB - OBJECTIVE: Cardiac valvular pathologies are frequently encountered as mechanical and functional disorders due to the calcification of the valves whatever the etiologies are. This pathophysiologic table usually ends up with valvular replacement. In this study, we aimed to decrease/eliminate the calcium in the excised calcified human heart valves by using citric acid in vitro hence bringing about the question for possible oral treatment of calcification of the valves by citric acid ingestion. METHODS: Fourteen pieces of mitral and/or aortic valves excised from 12 patients undergoing valve replacement were placed in a freshly prepared phosphate buffered saline solution containing 0.625% glutaraldehyde at +4 0C for 48 h. They were rinsed with 0.9% NaCl and divided into two groups; study and control. Control tissues were further treated in a freshly prepared solution with identical properties for another 5 days. Study tissues were placed into a solution containing 3.8% citric acid (pH 7.4) and kept for 48 h at +37 degrees C, then rinsed with 0.9% NaCl and transferred into a fresh solution containing 0.625% glutaraldehyde with phosphate buffer at 37 0C for 3 more days. Specimens were biochemically and histopathologically evaluated and compared using Mann Whitney U test. RESULTS: Calcium and phosphate levels in the study group were lower than in the control group (852.5+/-913.41 microg g-1 vs 413.05+/ 519.53 microg g-1, p=0.001 and 207.6+/-321.86 microg g-1 vs 124.4+/-289.48 microg g-1, p=0.035, respectively). Malondialdehyde and protein level values were changed insignificantly in the control and study groups. Histopathologic evaluation showed that collagen and elastin fibers were similar in both groups. In the study group, irregular and fusiform calcific formations around the collagen fibers were significantly decreased. CONCLUSIONS: Decalcifying human heart valves in vitro conditions with citric acid without an adverse change to the morphology of the valvular tissue specimens is meaningful. We believe that forwarding and looking for the answer to the question "whether systemic application of citric acid could lead to the decalcification and/or reduction of calcification in the native human heart valves" would be expressive. PMID- 18400628 TI - Statin use is associated with decreased CD-40 ligand expression on T lymphocytes of coronary atheroma plaque in patients with stable coronary artery disease. AB - OBJECTIVE: Atherosclerosis is a chronic inflammatory disease. Statins suppress the inflammation in the plaque. This cross-sectional study was planned to evaluate the effect of statins on plaque T cell activation markers in patients with stable angina pectoris undergoing coronary intervention and atherectomy procedures. METHODS: Twenty-six patients with stable angina with suitable for atherectomy coronary lesions were enrolled in the study. Fourteen of 26 patients who had been taking statin treatment for at least six months were assigned to the Group 1 (Statin group) and 12 patients who had not received any lipid lowering treatment comprised the Group 2 (Control group). Atherectomy specimens were studied with single and double immunohistochemical staining (CD25, CD69, and CD40L). Statistical analysis was performed using Student's t-test and Fisher's exact test. RESULTS: There was no significant difference between the total tissue area of sections (Group 1: 8.4+/-0.9 mm2, Group 2: 7.8+/-0.9 mm2, p>0.05). CD3, CD25, CD69, and CD40L positive cells did not show statistically significant difference between the groups in unit area (mm2). There was no significant difference between the groups for percentage of T lymphocytes expressing CD25 (Group 1: 7.8+/-4.6%, Group 2: 7.8+/-5.9%, p=0.97) and CD 69 (Group 1: 12.9+/ 4.6%, Group 2: 15.5+/-5.2%, p=0.203). The expression of CD40L was significantly lower in Group 1 than in Group 2 (Group 1: 4.8+/-3.9%, Group 2: 11.2+/-8.7%, p=0.034). CONCLUSION: We concluded that, statin treatment may decrease the expression of CD40L on plaque T lymphocytes in patients with stable angina pectoris. PMID- 18400629 TI - [Clonidine prevents development of hypertension in N (omega)-nitro-L-arginine treated rats]. AB - OBJECTIVE: Although, there are some evidences on the contribution of increased sympathoadrenergic activity on long-term nitric oxide synthase (NOS) inhibition induced hypertension, the contribution of sympathetic activity to the development of this model of hypertension are not sufficiently studied. The aim of the present study is to investigate the effects of clonidine on blood pressure and vascular alpha-adrenergic receptors in the long-term N (omega)-nitro-L-arginine (L-NNA) treated rats. METHODS: Sixty two Wistar rats were randomly divided into 8 groups. All groups were administrated L-NNA and/or clonidine in two different concentrations for ten days. L-NNA was administrated in concentrations of 15 and 45 mg/100ml to L-NNA15 and L-NNA45 groups, respectively. Clonidine was also administrated in concentrations of 150 and 225 microg/100ml to KLO150 and KLO225 groups, respectively. Blood pressure and heart rates were measured with tail-cuff method and plasma NOx levels with spectrophotometer. The a-adrenoreceptors responses were evaluated in thoracic aorta rings in "in vitro" conditions. RESULTS: Clonidine prevented the L-NNA induced hypertension dose-dependently, but did not effect the heart rates decreased by L-NNA. The heart rates and blood pressure of normotensive rats were not changed by clonidine alone. Plasma NOx levels increased in L-NNA15 group (0.62+/-0.11 micromol/L, p=0.003) but did not change in other groups. The sensitivity of aorta to phenylephrine (-7.33+/-0.11 micromol/L, p=0.001) and clonidine (-7.60+/-0.27 micromol/L, p=0.003) in L-NNA45 group and phenylephrine (-6.94+/-0.13 micromol/L, p=0.002) in L-NNA15 group increased. The sensitivity of aorta to phenylephrine (7.93+/-0.16 micromol/L, p=0.001) in KLO225 group and to clonidine (-7.20+/-0.10 micromol/L, p=0.009) in KLO150 group increased. CONCLUSION: This study supports the idea suggesting that symphathetic nervous system activation is partly responsible for the development of the long-term NOS inhibition induced hypertension. In conclusion, it was shown for the first time that clonidine prevents the development of long-term NOS inhibition induced hypertension dose-dependently. PMID- 18400631 TI - Mentholated cigarette smoking induced alterations in left and right ventricular functions in chronic smokers. AB - OBJECTIVE: Possible acute effects of smoking mentholated cigarette on left and right ventricular function is not known. The aim of the study was to compare acute effects of normal and mentholated cigarettes smoking on both ventricular diastolic functions in chronic smokers. METHODS: In a single-blinded, cross-over, open label and controlled study acute effect of smoking of the mentholated and the regular cigarettes was evaluated. Eighteen other than healthy regular cigarette smokers and 20 nonsmoker control subjects were included into the study. To compare the acute effects of mentholated and regular cigarette in each subject echocardiographic examination including tissue Doppler imaging (TDI) were performed at baseline, than in the smokers group TDI was measure 20-30 minutes after smoking two either cigarettes. RESULTS: In response to smoking two cigarettes, mitral E/A values declined from 1.78+/-0.44 to 1.58+/-0.41 after the regular cigarette (p=0.0043) and from 1.78+/-0.44 to 1.53+/-0.40 after the mentholated cigarette (p=0.0035). Tricuspid E deceleration time values declined from 185.28+/-20.05 ms to 222.72+/-26.47 ms after the regular cigarette (p<0.001) and 185.28+/-20.05 ms to 241.53+/-47.63 ms after the mentholated cigarette (p<0.001). Smoking of mentholated cigarette, but not regular cigarette smoking, increased tricuspid E deceleration time and right ventricular isovolumic contraction time (p=0.044; p=0.024 respectively) and decreased the right ventricular Em values (p=0.027). CONCLUSION: Mentholated and regular cigarette smoking have acute detrimental effects on right and left ventricular systolic and diastolic function. Mentholated cigarettes cause additional unfavorable acute effects on especially right ventricular tissue Doppler velocities, relaxation and contraction indices compared to regular cigarettes. PMID- 18400630 TI - Increased lipoprotein(a) in metabolic syndrome: is it a contributing factor to premature atherosclerosis? AB - OBJECTIVE: It is well known that patients with metabolic syndrome (MS) have a greater risk of developing coronary artery disease (CAD). However, the association of novel coronary risk factors with MS has not been well established. In this study, we sought to investigate the association of lipoprotein (a) [Lp(a)], homocysteine (Hcy), uric acid, and C-reactive protein (CRP) levels with MS. METHODS: We enrolled 355 consecutive patients from our outpatient cardiology clinic into this cross-sectional, controlled study-186 with MS and 169 without MS, according to the Adult Treatment Panel III criteria. Serum Hcy, Lp(a), uric acid, and CRP levels were determined and compared between the groups. RESULTS: The groups were homogenous with regard to age, sex, and other demographic variables (all p>0.05). As expected, the prevalence of hypertension (85.4% vs 55.6%, p<0.001) and dyslipidemia (78.3% vs 62.6%, p<0.05) were higher in patients in the MS group. Patients were comparable with respect to smoking (28.4% vs 24.8%, p =0.4) and family history of CAD (46.1% vs 40.8%, p=0.3). Patients with MS had significantly higher Lp(a) levels [29.2 (13.4-45.7) vs 16.2 (9.5-26.2) mg/dL; p<0.0001] compared with controls, whereas Hcy (12.2+/-4.8 vs 12.3+/-4.9 micromol/L; P=0.8), uric acid (5.7+/-1.6 vs 5.3+/-1.3 mg/dL; p=0.08), and CRP levels [6.0 (3.7-9.3) vs 5.1 (3.2-7.6) mg/L; p=0.07] were similar. CONCLUSION: Patients with MS seems to have increased serum levels of Lp(a), which might contribute to the premature atherosclerosis observed in these patients. Further research is needed to better clarify this issue. PMID- 18400632 TI - Clinical and angiographic importance of right bundle branch block in the setting of acute anterior myocardial infarction. AB - OBJECTIVE: To investigate functional status of patients (Killip class), left ventricular contractility, angiographic anatomy and severity of coronary lesions in patients with and without right bundle branch block (RBBB) in the setting of anterior myocardial infarction (MI). METHODS: Patients who admitted to coronary care unit with the diagnosis of acute anterior MI between 1999 and 2005 were retrospectively searched from our database. Out of 792 patients, 37 had RBBB (RBBB group) either at admission or in the course of anterior MI. Forty patients who developed no intraventricular conduction defect during the course of anterior MI with the same demographic characteristics were selected as the control group. RESULTS: Out of 37 patients, 30 had RBBB on admission and 7 developed RBBB in the course of acute MI. Left anterior descending artery (LAD) proximal lesion was more commonly detected in the RBBB group [23 (62.2%) vs. 11 (27.5%) patients, p=0.003]. Left ventricular ejection fraction was lower (33.0+/-4.2% vs 36.7+/ 4.9%, p=0.003) and end-systolic volume was higher (84.1+/-24.9 ml vs 74.6+/-22.0 ml, p=0.012) in patients with RBBB. Number of patients with high Killip grade (III and IV) was more in the RBBB group [7 (18.9%) vs 3 (7.5%), RR: 1.75, %95 CI 0.92-3.32, p=0.14], and number of patients with Killip grade I was more in the control group [34 (85.0%) vs 22 (59.5%), p=0.012]. Besides mean Killip class was higher in the RBBB group (1.65+/-0.90 vs 1.25+/-0.67, p=0.03). Three patients (8.1%) in the RBBB group and 2 patients (5.0%) in the control group died during hospitalization (p=0.67). CONCLUSION: Left ventricular ejection fraction decreases and Killip grade increases in case of RBBB in the setting of acute anterior MI. Culprit lesion in patients with RBBB is more commonly a LAD proximal lesion and threatened myocardial tissue is larger in patients with RBBB. PMID- 18400633 TI - The diagnostic value of multislice computed tomography in evaluation of coronary artery disease in patients with left bundle branch block. AB - OBJECTIVE: Noninvasive diagnostic methods do not supply sufficient data for patients with left bundle branch block (LBBB) accompanied with coronary artery diseases (CAD). Therefore, generally coronary angiography is required for these patients. Our aim was to evaluate the diagnostic value of multislice spiral computed tomographic (MSCT) coronary angiography to detect CAD in patients with LBBB. METHODS: Sixty one patients (31 males, 30 females, mean age: 56+/-13 years) with LBBB who have determined stenosis higher than 50% in quantitative coronary angiography were included in the cross-sectional study. The MSCT coronary angiography was applied to the patients with a 16-detector MSCT scanner that has an electrocardiographic synchronization unit. Each coronary artery was evaluated segmentally in the images acquired from MSCT coronary angiographies and any detected stenosis higher than 50% was recorded. RESULTS: The data of 793 coronary artery segments achieved from MSCT coronary angiographies of 61 patients (13 segments for each patient) were compared with the results of conventional coronary angiographies of the same patients. When all the segments evaluated were included in this comparison, the diagnostic accuracy, sensitivity, specificity, positive and negative predictive values of MSCT coronary angiography to detect stenosis higher than 50% were 91%, 67%, 97%, 85% and 92%, respectively. Also, it was observed that, MSCT coronary angiography has 80% sensitivity and 90% specificity to detect, at least, one segment coronary artery stenosis. CONCLUSION: The MSCT coronary angiography can be utilized as a noninvasive diagnostic method for patients with LBBB, in order to evaluate coronary artery disease. PMID- 18400634 TI - [Endovascular treatment of aortic lesions using the Medtronic Talent system: single center experience with mid-term follow-up]. AB - OBJECTIVE: The aim of this retrospective study is to investigate the safety and efficacy of endovascular repair of aortic lesions with the Medtronic Talent stent graft system and to present mid-term results of endovascular aortic repair performed in our center. METHODS: Between December 2002 and March 2007, 54 patients (6 women) with aortic (14 thoracic and 40 abdominal) lesions underwent treatment with Talent stent-graft. The average age of the patients was 64.8 (20 88) years. Duration of follow-up period ranged from 1 to 49 months (average 21 months). Indications for endovascular repair were degenerative aneurysm in 45, degenerative aneurysm and penetrating ulcer in 2, only penetrating ulcer in 1, traumatic thoracic isthmic transsection in 4 and vasculitic aneurysm secondary to Behcet's disease in 2 patients. RESULTS: Repair was performed with the tubular (16), aortouniiliac (1) or bifurcated (37) stent-grafts. Technical success rate was 100%. No death, major complication or need of immediate conversion to open repair was seen. Endoleak rate was 18.5% at 1 month follow-up period. Thirty-day mortality was 1.8% and morbidity (other than endoleaks) rate was 12.9%. During the follow-up period, secondary intervention was required in 12.9% of patients. Iliac limb occlusion was detected in 1 patient (1.8%). Graft migration causing type 1 endoleak and requiring open surgical treatment was seen in 1 patient (1.8%). Four patients (7.4%) are still under follow-up for type-2 endoleaks that do not require intervention. No graft infection or death due to aneurysm rupture was detected. CONCLUSION: Endovascular treatment of aortic lesions in selected patients with comorbid conditions using the Talent stent-graft exhibits a high degree of technical success with a low perioperative morbidity and mortality rate. The major disadvantage of endovascular aortic repair is necessity of life long imaging follow-up and secondary interventions. PMID- 18400635 TI - Human soluble tumor necrosis factor receptor I (sTNF-RI) and interleukin-I receptor antagonist (IL-I Ra) in different stages of acute rheumatic fever. AB - OBJECTIVE: Acute rheumatic fever (ARF) results from an autoimmune response to infection with group A streptococci. Serum concentrations of two anti inflammatory cytokines, interleukin-I receptor antagonist (IL-IRa) and human soluble tumor necrosis factor receptor I (sTNF-RI) were determined in patients with ARF at the time of admission and 3 months after treatment in order to evaluate changes in cytokine concentrations occurring during different stages of the disease. METHODS: Serum concentrations of two anti-inflammatory cytokines, IL I Ra and sTNF-RI , were investigated in children with ARF at the time of admission (n=21) and after 3 months following the cessation of treatment (n=15). The sTNF-RI and sIL-IRa were measured quantitatively in serum using enzyme-linked immunosorbent assay (ELISA). RESULTS: Levels of IL-1Ra and sTNF-RI were found to be significantly higher during acute phase and remission period of ARF when compared to age-matched healthy controls (p=0.001 and p=0.0001, respectively). CONCLUSION: Our study demonstrated that two anti-inflammatory cytokines, serum sTNFRI and IL-1Ra, are increased in acute and remission stages of ARF reflecting activation of the cellular immune response. We suggest this increase might probably be generated in an effort to counteract the already increased concentrations of proinflammatory cytokines. PMID- 18400636 TI - [Myocardial performance index]. AB - Myocardial performance index (MPI) is a numeric value, which could be obtained by using cardiac time intervals. This numeric value is defined as the sum of isovolumetric contraction time (ICT) and isovolumetric relaxation time (IRT) divided by ejection time (ET) and could be calculated for each ventricle individually. Myocardial performance index has been invented as a Doppler index of combined systolic and diastolic myocardial performance in patients with primary myocardial systolic dysfunction. Myocardial performance index has prognostic value in various clinical settings seems to be independent of heart rate. In this review, clinical applications of MPI are scrutinized. PMID- 18400637 TI - [Global confusion on the diagnostic criteria for metabolic syndrome: what is the point that guidelines can not agree?]. AB - Several international and national associations have proposed their own diagnostic criteria for metabolic syndrome. Regarding the heterogeneity of these guidelines, some authors refused the usage of metabolic syndrome definition. Here we aimed to analyze the differences between the diagnostic criteria proposed for metabolic syndrome. We reviewed and compared these different guidelines' criteria. We conclude that, despite the heterogeneity, there is a conceptual agreement on the definition of metabolic syndrome. But cut-off values and selection of diagnostic parameters are still conflicting. Diagnostic or definitive components of the syndrome (i.e., obesity, insulin resistance) should be considered separately from the associated-conditions (i.e., polycystic ovary, obstructive sleep apnea, microalbuminuria, non-alcoholic steatohepatitis etc.) of metabolic syndrome, during the course of diagnosis. Maintenance of the metabolic syndrome as a diagnostic category would still seem to be useful for an effective multiple cardiovascular risk prediction. PMID- 18400638 TI - [Thromboelastography and its use in cardiac surgery]. AB - Thromboelastography is an alternative method to conventional coagulation tests for the general evaluation of hemostatic system. Cardiac surgery with cardiopulmonary bypass is accomplished by complex alterations of hemostasis, including acquired dysfunction of platelets, consumption coagulopathy and increased fibrinolysis. Despite major advances in blood conservation methods and perioperative care of the patients, transfusion rates in cardiac surgery remain high. Thromboelastography has an ability to assess almost all components of haemostatic system globally. Currently, thromboelastography is used with standard coagulation tests to decrease the microvascular bleeding and homologous blood transfusion in cardiac surgery with cardiopulmonary bypass. In this review, we aimed to discuss thromboelastography technology and its usage in cardiac surgery. PMID- 18400639 TI - Implantation of a cardioverter defibrillator on the side of a hemodialysis fistula to prevent loss of the alternative arterio-venous access. PMID- 18400640 TI - Persistent atrial fibrillation associated with gastroesophageal reflux accompanied by hiatal hernia. PMID- 18400641 TI - A complex congenital cardiovascular anomaly: persistence of left inferior and superior caval veins in conjunction with atrial and ventricular septal defects. PMID- 18400642 TI - [Tricuspid valve mass: magnetic resonance imaging findings--case report]. PMID- 18400643 TI - Midterm survival following repair of a giant left ventricular true aneurysm ruptured during operation and associated with papillary muscle rupture. PMID- 18400644 TI - [Implantation of pulmonary valve without use of cardiopulmonary bypass: an innovative method. Case report]. PMID- 18400645 TI - The experience with the Epiclose-T vascular access closure device: a human study. PMID- 18400646 TI - Can a city population represent a whole country? Essentials of study design for epidemiologic studies/ Serum lipid profiles including non-high density lipoprotein cholesterol levels in Turkish school-children. PMID- 18400647 TI - An isolated case of left ventricular non-compaction with sick sinus syndrome. PMID- 18400649 TI - Original Images. Giant paraanastomotic iliac artery pseudoaneurysm. PMID- 18400648 TI - Original Images. Posttraumatic pneumopericardium and bilateral pneumothorax. PMID- 18400651 TI - Health evaluation of gold miners living in a mercury-contaminated village in Serra Pelada, Para, Brazil. AB - Serra Pelada is a village in the Amazon region of Brazil where most of the inhabitants are former gold miners. Of 235 individuals evaluated, 219 were males (93.19%), 16 were females (6.80%), and the mean age was 52.07 years (standard deviation = 11.57). Most were heavy drinkers (62.44%) and smokers (70.30%), and 85.53% had previously suffered from malaria. Reported symptoms included fatigue (30.60%), irritability (35.62%), excitability (14.16%), insomnia (34.48%), memory loss (61.80%), visual field constriction (4.18%), paresthesia (64.93%), partial hearing loss (16.35%), and gingivitis (18.01%). After an examination of the residents, the authors observed several neurological symptoms: tremors (22.80%), involuntary ocular movement (2.20%), visual field constriction (4.18%), Romberg syndrome (2.33%), involuntary tongue movement (2.19%), dysdiadochokinesia (0.43%), failure of a finger-nose test (10.96%), failure of a knee-heel test (4.84%), inability to complete a tandem march (6.25%), muscular weakness (2.27%), and damage to sensory organs (24.66%). The authors concluded that these neurological changes possibly resulted from mercury toxicity; however, they could not determine a significant correlation with the mercury levels detected in participants' urine. PMID- 18400653 TI - Cigarette smoking and the risk of adult leukemia: results from the Three Mile Island cohort study. AB - Smoking is an unconfirmed risk factor for the development of leukemia. The authors examined the potential link using data from the Three Mile Island cohort for the period 1979-1995. Eligible for analysis were 24,539 individuals aged 14 years or older who were followed up over 16 years from the Three Mile Island cohort. The authors identified all incident leukemia cases through the Pennsylvania Department of Health Cancer Registry. They used the Cox proportional hazards model to evaluate the relationships and observed 42 incident leukemia cases, including 15 acute myeloid leukemia (AML) cases, in the cohort. After controlling for other confounding factors, the authors found current smoking to be associated with an increased risk of adult AML (relative risk = 3.47; 95% confidence interval = 1.002-11.99). The authors also observed a marginally significant linear trend of risk of AML associated with the number of years smoked (p = .06). The results from this study suggested that cigarette smoking was associated with an increased risk of adult AML. Further investigation is required to confirm these findings. PMID- 18400654 TI - Exposure dose and significance of platinum and platinum salts in breast implants. AB - The objectives of this study were to illustrate and inform key scientific issues, as determined from the peer-reviewed literature, that are critical to developing an accurate understanding of the current state of knowledge regarding platinum (Pt) in medical devices (ie, breast implants). The author identified most studies for inclusion via a PubMed database search; she extracted descriptive statistics from the studies. The author calculated Pt and Pt salt exposure doses for environmental and occupationally related samples. She observed that a number of samples elicited biological effects over a wide range of concentrations. A single silicone breast implant may be expected to contain higher Pt and Pt salt doses than have produced adverse health effects in humans. The author posits a biologically plausible rationale for Pt salt-related health problems in women that have been exposed to silicone breast implants. PMID- 18400655 TI - Sick building syndrome: psychological, somatic, and environmental determinants. AB - The authors aimed to examine potential relationships between work-related symptoms attributed to sick building syndrome (SBS) and certain psychological, somatic, and environmental factors. The multidisciplinary, cross-sectional study comprised 171 female subjects working in air-conditioned and naturally ventilated nonindustrial office buildings. The authors collected information concerning symptoms related to SBS and made assessments of quality of life by using appropriate questionnaires. They assessed the women's levels of emotional stability or neuroticism using the Cornell Index. They determined skin and airway reactivity markers and indoor microclimate data by using standardized methods. The study showed that the subjects had a high prevalence of fatigue (60.2%), sore and dry eyes (57.9%), and headache (44.4%), as well as a generally high score according to the SBS Index. Neuroticism and subjectively estimated physical health as well as the type of building ventilation significantly contributed to the prediction of the SBS Index, explaining 15% of the variance. PMID- 18400656 TI - Probable occupational endophthalmitis from Bacillus cereus. AB - Endophthalmitis is a severe infection produced by the introduction of microorganisms into the eye after penetrating injury, surgery, or hematogenous spread from a distant primary site of infection. The case presented is a 44-year old man who worked as a machine operator with exposure to substantial metalworking fluid aerosols from a high-speed grinder generating fine particles. PMID- 18400657 TI - Reduction in telomere length in individuals exposed in utero to glycol ether. AB - Little is known about the mechanism by which ethylene glycol monomethyl ether (EGME) produces genotoxic effects in humans. The authors found that individuals exposed in utero to EGME showed characteristic dysmorphic features, unexplained mental retardation, and persistent cytogenetic damage. They hypothesized that these individuals had a higher level of terminal chromosome arrangements, accounting for the genomic instability detected. Results indicate that in utero exposed individuals have significantly reduced telomeres relative to non-in utero exposed participants. The M +/- SEM pixel intensity in the in utero-exposed participants was 43.6 +/- 7.6 compared with 74.1 +/- 1.9 in the non-in utero exposed. Findings suggest that exposure to EGME in utero could result in terminal chromosome rearrangements and shortening of telomere length, leading to the observed dysmorphic features and idiopathic mental retardation. PMID- 18400658 TI - American College Health Association - National College Health Assessment spring 2007 reference group data report (abridged). AB - OBJECTIVE: Assessing and understanding the health needs and capacities of college students is paramount to creating healthy campus communities. The American College Health Association-National College Health Assessment (ACHA-NCHA) is a survey that ACHA developed in 1998 to assist institutions of higher education in achieving this goal. The ACHA-NCHA contains approximately 300 questions assessing student health status and health problems, risk and protective behaviors, and impediments to academic performance. PARTICIPANTS: The spring 2007 reference group includes ACHA-NCHA data from 71,860 students at 107 institutions of higher education. METHODS: Officials at participating institutions administered the ACHA NCHA to all students, to randomly selected students, or to students in randomly selected classrooms. ACHA collected data between January and May 2007. RESULTS: Results from the spring 2007 reference group (N= 71,860) are presented. CONCLUSIONS: These data expand the understanding of the health needs and capacities of college students. PMID- 18400659 TI - Wired: energy drinks, jock identity, masculine norms, and risk taking. AB - OBJECTIVE: The author examined gendered links among sport-related identity, endorsement of conventional masculine norms, risk taking, and energy-drink consumption. PARTICIPANTS: The author surveyed 795 undergraduate students enrolled in introductory-level courses at a public university. METHODS: The author conducted linear regression analyses of energy-drink consumption frequencies on sociodemographic characteristics, jock identity, masculine norms, and risk-taking behavior. RESULTS: Of participants, 39% consumed an energy drink in the past month, with more frequent use by men (2.49 d/month) than by women (1.22 d/month). Strength of jock identity was positively associated with frequency of energy-drink consumption; this relationship was mediated by both masculine norms and risk-taking behavior. CONCLUSIONS: Sport-related identity, masculinity, and risk taking are components of the emerging portrait of a toxic jock identity, which may signal an elevated risk for health-compromising behaviors. College undergraduates' frequent consumption of Red Bull and comparable energy drinks should be recognized as a potential predictor of toxic jock identity. PMID- 18400660 TI - Prevalence and correlates of self-injury among university students. AB - OBJECTIVE: The authors' purpose in this research was to establish estimates of the prevalence and correlates of nonsuicidal self-injury among university students. PARTICIPANTS: The authors recruited participants (N = 2,843) from a random sample of 5,021 undergraduate and graduate students attending a large midwestern public university. METHODS: Using an Internet-based survey, the authors measured the prevalence of self-injury and potential risk factors, including depression, anxiety, eating disorders, suicidal thoughts, and negative health behaviors. RESULTS: Seven percent of students reported any self-injury over the previous 4 weeks. Factors associated with a significantly higher likelihood (p <.05) of self-injury included cigarette smoking, concurrent depressive and anxiety disorders, and for men, growing up in a family of low socioeconomic status and having symptoms of eating disorders. Only 26% of those who reported self-injury received mental health therapy or medication in the previous year. CONCLUSIONS: Students who harm themselves experience high anxiety and distress, yet are unlikely to seek help. PMID- 18400661 TI - Anabolic androgenic steroids: use and perceived use in nonathlete college students. AB - OBJECTIVE: The authors investigated the use and perceived use of anabolic androgenic steroids (AAS) among nonathlete college students. PARTICIPANTS: The authors surveyed a sample of 485 nonathlete college students at a major metropolitan university. METHODS: They administered a survey on use and perceived use of AAS to the students. RESULTS: Forty-two participants (9%) reported using AAS (37 men, 5 women). Seniors were the most likely to use AAS (36%), and freshman the least likely (7%). Thirty-four percent of nonusers and 41% of users indicated they knew between 1 and 5 AAS users. Of the total sample, 36% perceived that 5% to 10% of nonathlete college students used AAS. Reasons for AAS use were because friends were using (7%), a desire to enhance physical appearance (45%), and a desire to increase physical performance (48%). CONCLUSIONS: These findings may have significant implications in planning strategic preventive educational programs, and health educators should target incoming college freshmen with the intent of dissuading AAS use. PMID- 18400662 TI - Energy expenditure during physically interactive video game playing in male college students with different playing experience. AB - OBJECTIVE: Researchers have yet to explore the effect of physically interactive video game playing on energy expenditure, despite its potential for meeting current minimal daily activity and energy expenditure recommendations. PARTICIPANTS AND METHODS: Nineteen male college students -- 12 experienced Dance Dance Revolution (DDR) players and 7 inexperienced players -- completed maximal oxygen uptake assessments and a 30-minute DDR gaming session. The authors recorded heart rate (HR), rating of perceived exertion (RPE), respiratory exchange rate (RER), oxygen consumption (VO2), and total steps (TS30). RESULTS: Experienced participants showed higher exercise HR, RPE, RER, VO2, total and relative energy expenditure, exercise intensity, TS30, and average steps per minute, and less time and steps to expend 150 kilocalories (p < .05). CONCLUSIONS: Participants with greater playing experience can work at higher intensities, promoting greater energy expenditure. PMID- 18400663 TI - College students' gambling behavior: when does it become harmful? AB - OBJECTIVE: The authors investigated behavioral indicators of pathological gambling in a college student sample. PARTICIPANTS AND METHODS: The authors administered a diagnostic interview for pathological gambling to 159 college students, who also completed a demographic questionnaire, and a self-report measure of psychological distress. RESULTS: Consistent patterns of gambling behavior associated with pathological gambling included gambling more than 1.2 times per month, gambling more than 2.1 hours per month, intending to wager more than 6.1% of monthly income, and wagering more than 10.5% of monthly income. In addition, the combination of psychological distress, gambling frequency, and a history of parental gambling problems was significantly associated with pathological gambling in college students. CONCLUSIONS: These findings suggest initial markers of problematic gambling behavior in a college sample that may be used in assessment, prevention, and intervention. PMID- 18400664 TI - Behavior change and the freshman 15: tracking physical activity and dietary patterns in 1st-year university women. AB - OBJECTIVE AND PARTICIPANTS: The authors assessed the stability of diet and physical activity and their relationship to weight changes in first-year university women. METHODS: They collected anthropometric and body composition data from 101 resident women at the beginning of their first year of college and again at 12 months. The authors obtained physical activity and dietary logs 4 times throughout the year. RESULTS: Caloric intake decreased over 12 months in all participants (p = .01). There was little change in physical activity in participants who lost weight (p = .73, d = .18). Those who gained weight experienced a trend toward decreased physical activity (p = .13, d = .38). A significant Time X Group interaction on physical activity (p = .04) suggests that physical activity patterns differed substantially between individuals who gained weight and those who lost weight. CONCLUSIONS: Reduction in physical activity appears to be the defining characteristic in freshman weight gain. PMID- 18400666 TI - Over-the-counter medication and herbal or dietary supplement use in college: dose frequency and relationship to self-reported distress. AB - OBJECTIVE: A growing number of researchers have examined the use of over-the counter (OTC) medications and herbal or dietary supplements among college students. There is concern about the efficacy and safety of these products, particularly because students appear to use them at a higher rate than does the general public. PARTICIPANTS AND METHODS: The authors administered surveys to college students (N = 201) to assess the frequency of use in the past week. RESULTS: A substantial percentage reported using OTC medications (74.1%), herbal or dietary supplements (70.6%), or both concurrently (61.2%). Dose frequency of OTC medications was the best predictor of self-reported emotional distress in the past week. Higher doses of products containing pseudoephedrine or valerian were associated with self-reported anxiety. CONCLUSIONS: These data further reflect an increasing trend toward self-medication among college students. Investigators must conduct reliability and validity studies to evaluate the clinical utility of the measurement tool developed in this study. PMID- 18400665 TI - The Freshman 15: is it real? AB - OBJECTIVE: The belief that college students gain 15 lbs during freshman year is widespread, yet the evidence for this is limited. The authors aimed to determine whether college students gain weight during freshman year. PARTICIPANTS: The authors studied unmarried freshmen living on-campus at a private university in the northeastern United States. METHODS: The authors used an online survey to collect information about social behaviors and weight. RESULTS: The authors observed an average weight gain of 2.7 lbs. About half of the students gained weight, and 15% lost weight. Men gained more weight than did women. CONCLUSIONS: Freshman weight gain was 5.5 times greater than that experienced by the general population. PMID- 18400668 TI - Indoor tanning and problem behavior. AB - OBJECTIVE: The authors examined factors predicting college students' use of tanning beds. PARTICIPANTS AND METHODS: Undergraduate students (N = 745) at a large Northeastern university participated in the study by answering a survey measuring tanning behavior and other psychosocial variables, including sensation seeking, self-esteem, tanning image beliefs, and friends' tanning bed use. RESULTS: All 3 systems from problem behavior theory predicted past tanning bed use and intention to use tanning beds. The authors observed a positive association between sensation seeking and intention to use tanning beds. Tanning image beliefs were positively associated with both past tanning behavior and intention to use tanning beds. CONCLUSIONS: Interventions focusing on friend and acquaintance social network influences may be more effective than health-risk campaigns in reducing tanning bed use. PMID- 18400667 TI - Assessing college students' autonomy over smoking with the Hooked On Nicotine Checklist. AB - OBJECTIVE, PARTICIPANTS, AND METHODS: In this study, the authors explored the psychometric properties of the Hooked on Nicotine Checklist (HONC) among 300 college students who were current smokers. The HONC is a 10-item survey instrument designed to measure diminished autonomy over smoking, a key aspect of dependence. Autonomy is diminished when symptoms present a barrier to cessation. RESULTS: Internal consistency was high (alpha = .89), as was concurrent validity. Students who began smoking earlier and heavier smokers reported more symptoms than did those who started later and smoked less. After controlling for smoking frequency, the HONC was predictive of the likelihood of a failed cessation attempt, with each additional symptom doubling that likelihood. CONCLUSIONS: College health professionals could use the HONC to help new smokers recognize their symptoms of diminished autonomy. The HONC may serve as a recruiting tool for cessation programs, which could benefit such early-phase smokers. PMID- 18400669 TI - Sleep patterns of college students at a public university. AB - OBJECTIVE: The authors' purpose in this study was to determine the sleep patterns of college students to identify problem areas and potential solutions. PARTICIPANTS: A total of 313 students returned completed surveys. METHODS: A sleep survey was e-mailed to a random sample of students at a North Central university. Questions included individual sleep patterns, problems, and possible influencing factors. RESULTS: Most students reported later bedtimes and rise times on weekends than they did on weekdays. More than 33% of the students took longer than 30 minutes to fall asleep, and 43% woke more than once nightly. More than 33% reported being tired during the day. The authors found no differences between freshmen, sophomores, juniors, seniors, and graduate students for time to fall asleep, number of night wakings, or total time slept each night. CONCLUSIONS: Many students have sleep problems that may interfere with daily performance, such as driving and academics. Circadian rhythm management, sleep hygiene, and white noise could ameliorate sleep difficulties. PMID- 18400670 TI - Are peer educators really peers? AB - OBJECTIVE, PARTICIPANTS, AND METHODS: To determine whether peer educators are considered peers by their audiences, the authors compared 28 peer educators and 28 college students on their personal qualities using self-report surveys. RESULTS: Students who wanted to become peer educators reported higher self esteem, greater leadership skills, and fewer risky health behaviors than did demographically similar college students. However, peer educators appeared similar to their colleagues in terms of personal values and personality temperament. CONCLUSIONS: Although peer educators are natural leaders with strong self-esteem and more healthy behaviors, they appear to be quite similar to their audiences in regard to values and temperaments, making them successful role models and arguably true peers. PMID- 18400671 TI - Meditation lowers stress and supports forgiveness among college students: a randomized controlled trial. AB - OBJECTIVE AND PARTICIPANTS: The authors evaluated the effects on stress, rumination, forgiveness, and hope of two 8-week, 90-min/wk training programs for college undergraduates in meditation-based stress-management tools. METHODS: After a pretest, the authors randomly allocated college undergraduates to training in mindfulness-based stress reduction (MBSR; n = 15), Easwaran's Eight Point Program (EPP; n = 14), or wait-list control (n = 15). The authors gathered pretest, posttest, and 8-week follow-up data on self-report outcome measures. RESULTS: The authors observed no post-treatment differences between MBSR and EPP or between posttest and 8-week follow-up (p > .10). Compared with controls, treated participants (n = 29) demonstrated significant benefits for stress (p < .05, Cohen's d = -.45) and forgiveness (p < .05, d = .34) and marginal benefits for rumination (p < .10, d = -.34). CONCLUSIONS: Evidence suggests that meditation-based stress-management practices reduce stress and enhance forgiveness among college undergraduates. Such programs merit further study as potential health-promotion tools for college populations. PMID- 18400672 TI - Weight loss practices and body weight perceptions among US college students. AB - OBJECTIVE: The authors assessed associations between body weight perception and weight loss strategies. PARTICIPANTS: They randomly selected male and female college students (N = 38,204). METHODS: The authors conducted a secondary data analysis of the rates of weight loss strategies and body weight perception among students who completed the National College Health Assessment survey. RESULTS: Half of respondents (50%) were trying to lose weight, although only 28% of students were overweight or obese. Also, 12% of respondents had inaccurate body weight perception. Women and men with inaccurate body weight perception were significantly more likely to engage in inappropriate weight loss strategies than were those with accurate body weight perception. Of all students attempting to lose weight, 38% used both diet and exercise. CONCLUSIONS: These data show that college students are interested in weight loss and that body weight perception plays an important role in the desire to lose weight. PMID- 18400673 TI - Campus walkability, pedometer-determined steps, and moderate-to-vigorous physical activity: a comparison of 2 university campuses. AB - OBJECTIVE AND PARTICIPANTS: At 2 Arizona State University (ASU) campuses, the authors measured student activity and distance walked on campus, as well as student-reported walkability around the student union. METHODS: Students from ASU Polytechnic (n = 20, 33% male) and ASU-Tempe (n = 20, 60% male) recorded distance walked on campus and wore physical activity monitors for 5 days. RESULTS: Polytechnic students spent an average of 36.9 minutes in moderate-to-vigorous physical activity each week; Tempe students spent 69.5 minutes (p < .001). At Polytechnic, students walked an average of 7,674 steps per weekday; at Tempe, 11,294 steps (p = .003). Female students at Polytechnic walked an average of 1.3 km/d; at Tempe, 4.3 km/d (p < .001). At Polytechnic, men walked an average of 1.4 km/d; at Tempe, 3.1 km/d (p = .03). Tempe students rated campus walk-ability as very good, whereas Polytechnic students rated it fair (p < .001). CONCLUSIONS: Students at both campuses met activity recommendations; noted differences may be attributed to the built environment's contribution to walkability. PMID- 18400674 TI - Tattoos can harm perceptions: a study and suggestions. AB - OBJECTIVE: Health researchers have claimed that perceptions toward a person with a tattoo are more negative than are perceptions toward nontattooed persons. However, support for this has been obtained almost completely by nonexperimental research. PARTICIPANTS: In 2 experiments with 158 community college student participants, the authors found that tattoos harmed perceptions. METHODS: Students viewed a photograph of a female model with and without a visible tattoo, and rated her on 13 personal characteristics. RESULTS: In Experiment 1, ratings of a model with a dragon tattoo were significantly more negative (p < .05) on 5 of the 13 personal characteristics than were ratings of the same model shown without the tattoo. In Experiment 2, which included different participants, a different model, and a different tattoo, the authors found that a dolphin tattoo led to more negative ratings on 2 of the 13 characteristics. CONCLUSIONS: The authors discuss possible impacts of tattoos on person perception as well as implications of the results for college student healthcare providers. PMID- 18400675 TI - Evaluation of arm dominance by using the mechanomyographic signal. AB - The authors collected and compared mechanomyo-graphic (MMG) signal behavior from the biceps brachii of left and right arms of right-handed men (n = 19) and women (n = 20) who performed isometric contractions at 5 contraction levels. Mean frequency (MF) and RMS values were calculated from the MMG signals that arose from lateral oscillations of muscle fibers. Across genders and arms, RMS values increased with contraction level and MF values decreased with increase in muscle contraction. The authors found no significant difference in those parameters between dominant and nondominant arms. Therefore, summation of muscle twitches obtained from the MMG signal collected in biceps brachii muscle does not reveal any difference between dominant and nondominant arms. PMID- 18400676 TI - Interaction between the development of postural control and the executive function of attention. AB - The authors examined the interaction between the development of postural control and the development of the executive function of attention in 13 children and 6 adults in dual-task conditions. Participants performed an attentionally demanding cognitive task and a postural task simultaneously. The authors equalized the attentional load of the cognitive task across age groups. Comparative changes in the center of pressure in dual- and single-task conditions indicated that dual tasks interfered with postural performance in the wide stance (WS) and the modified Romberg stance (RS). Children at 4-6 years of age (but not children at ages 7-12 years of age or adults) experienced postural control interference in both stance positions, but interference was greater in the RS (p = .018). For all participants, cognitive task performance in RS was unchanged from that in WS. The knowledge gained from the results of this study will contribute to the design and implementation of academic and preacademic programming for young children. Their performance of an intentionally demanding cognitive task would be enhanced by the provision of appropriately sized desks and chairs or their use of an alternate, less demanding position. PMID- 18400677 TI - The role of stability in the dynamics of learning, memorizing, and forgetting new coordination patterns. AB - According to a dynamic theory of learning, how a new memory is formed depends on the stability of the nearest preexisting memories. To predict retention after practice, the authors analyzed how 15 participants memorized 2 bimanual coordination patterns (45 degrees or 135 degrees relative phase). The authors assessed (a) how participants memorized the required patterns with learning and (b) how the associated memory layout evolved. Results showed that a practiced 45 degrees pattern near a very stable memory (0 degrees ) persisted, whereas a 135 degrees pattern near a less stable memory (180 degrees ) was forgotten. Those findings corroborate the proposition that retention of coordination patterns depends on the stability of the extant motor memories. The authors discuss that proposal in terms of the coevolution of accuracy and stability with learning to predict persistence of required or false memories. PMID- 18400678 TI - Effect of imagined movement speed on subsequent motor performance. AB - Researchers realize that motor imagery (MI) duration is closely linked to actual motor action duration. In 2 experiments, the authors investigated the effect of changing MI speed on actual movement duration over a 3-week training period. Experiment 1 involved 2 series of body movements that 24 participants mentally performed faster or slower than their actual execution speeds. The fast MI group's actual times decreased on subsequent performance. Participants in Experiment 2 were 21 skilled athletes who increased (decreased) their well rehearsed actual movement times after MI training at a slow (fast) speed. The effect was task-related, however: MI affected only self-initiated movement. The effect of MI on actual speed execution supports the ideomotor theory because anticipation of sensory consequences of actions is mentally represented. PMID- 18400679 TI - Constrained motor-perceptual task in infancy: effects of sensory modality. AB - The authors examined the relative success of varying sensory stimulation modalities that they presented via a mobile reinforcement procedure for promoting left-knee extensions in 3-month-old infants. They separated 53 infants into 5 groups. Four groups received contingent unimodal auditory, enhanced auditory, or visual reinforcement or contingent bimodal auditory plus visual (aud + vis) reinforcement. One group (controls) received aud + vis noncontingent reinforcement. The group that received aud + vis contingent reinforcement was most successful in learning the motor task and maintained the highest attention levels. The authors observed few differences in learning and attention within the unimodal groups. The present findings confirm the effectiveness of contingent multisensory stimulation for promoting perception-action coupling in infancy. PMID- 18400680 TI - Extroversion-related differences in speed of premotor and motor processing as revealed by lateralized readiness potentials. AB - To further elucidate extroversion-related differences in speed of sensorimotor processing, the authors obtained behavioral and psychophysiological measures as participants (16 introverts and 16 extroverts) performed a visual go/no-go task. Although no extroversion-related differences in reaction time emerged, introverts showed faster premotor processing but slower central and peripheral motor processing--as indicated by latencies of the lateralized readiness potential (LRP) and electromyographic (EMG) data, respectively--than extroverts did. Additional regression analyses revealed that stimulus-locked LRP latency, response-locked LRP latency, and Nl EMG amplitude accounted for 40% of overall variability in individual extroversion scores. On the basis of the present results, the authors introduce a compensation hypothesis that accounts for the common failure of researchers to demonstrate extroversion-related differences in reaction time. The present results challenge J. Brebner and C. Cooper's (1985) model of extroversion in which stimulus analysis is not slower in introverts than in extroverts. However, the present findings support the assumption of faster motor processing in extroverts. PMID- 18400681 TI - Structured perceptual arrays and the modulation of Fitts's law: examining saccadic eye movements. AB - On the basis of recent observations of a modulation of Fitts's law for manual pointing movements in structured visual arrays (J. J. Adam, R. Mol, J. Pratt, & M. H. Fischer, 2006; J. Pratt, J. J. Adam, & M. H. Fischer, 2007), the authors examined whether a similar modulation occurs for saccadic eye movements. Healthy participants (N = 19) made horizontal saccades to targets that appeared randomly in 1 of 4 positions, either on an empty background or within 1 of 4 placeholder boxes. Whereas in previous studies, placeholders caused a decrease in movement time (MT) without the normal decrease in movement accuracy predicted by Fitts's law, placeholders in the present experiment increased saccadic accuracy (decreased endpoint variability) without an increase in MT. The present results extend the findings of J. J. Adam et al. of a modulation of Fitts's law from the temporal domain to the spatial domain and from manual movements to eye movements. PMID- 18400682 TI - Short-term limb immobilization affects motor performance. AB - C. Ghez, J. Gordon, and M. R Ghilardi (1995; J. Gordon, M. R Ghilardi, & C. Ghez, 1995; R. L. Sainburg, M. R Ghilardi, H. Poizner, & C. Ghez, 1995) have found that proprio-ceptive deafferentation impairs feedforward and feedback mechanisms that control reaching movements. In the present study, the authors found immobilization-induced changes in limb kinematics, including joint motion, in 32 healthy participants who performed out-and-back movements before and after 0, 6, or 12 hr of immobilization of the left arm. Control participants did not undergo the arm immobilization procedure. Immobilization for 12 hr, but not 6 hr, caused trajectories with increased hand-path areas and altered interjoint coordination. The abnormalities were smaller in amplitude but similar in quality to those reported in deafferented patients (R. L. Sainburg et al.). In addition, movement onset point significantly drifted after immobilization. Thus, short-term limb disuse can affect interjoint coordination by acting on feedforward mechanisms. These behavioral alterations are potentially related to cortical plastic changes. PMID- 18400683 TI - Species detection using environmental DNA from water samples. AB - The assessment of species distribution is a first critical phase of biodiversity studies and is necessary to many disciplines such as biogeography, conservation biology and ecology. However, several species are difficult to detect, especially during particular time periods or developmental stages, potentially biasing study outcomes. Here we present a novel approach, based on the limited persistence of DNA in the environment, to detect the presence of a species in fresh water. We used specific primers that amplify short mitochondrial DNA sequences to track the presence of a frog (Rana catesbeiana) in controlled environments and natural wetlands. A multi-sampling approach allowed for species detection in all environments where it was present, even at low densities. The reliability of the results was demonstrated by the identification of amplified DNA fragments, using traditional sequencing and parallel pyrosequencing techniques. As the environment can retain the molecular imprint of inhabiting species, our approach allows the reliable detection of secretive organisms in wetlands without direct observation. Combined with massive sequencing and the development of DNA barcodes that enable species identification, this approach opens new perspectives for the assessment of current biodiversity from environmental samples. PMID- 18400684 TI - Personality as a predictor of dietary quality in spouses during midlife. AB - The authors evaluated the NEO Personality Inventory-Revised (NEO-PI-R) as a predictor of dietary quality in 850 married couples, focusing on associations among each participant's personality as a predictor of their own dietary quality and their spouses' dietary quality. Diet was based on a modified version of the US Department of Agriculture Healthy Eating Index. Openness was associated with self-ratings of dietary quality for wives (r = .28) and husbands (r = .27). Wives' levels of the characteristic openness were also related to their spouses' ratings of dietary quality (r = .22). The primary facets of openness accounting for the domain-level findings were O2-aesthetics and O4-actions. The remaining personality domains (neuroticism, extraversion, agreeableness, and conscientiousness) were not associated with self or spousal ratings of dietary quality (rs = .08-.09). Openness was associated with healthy eating habits- findings that may affect disease prevention during, midlife. PMID- 18400685 TI - Client encounters in alternative HIV testing sites: counselors' perceptions and experiences. AB - In this study, the authors explored HIV test counselors' perceptions of and experiences working with "difficult" and "good" clients in alternative HIV testing sites. Trained interviewers made field observations and conducted sixteen 60-minute, semistructured interviews with counselors. Counselors reported 7 main characteristics of difficult clients: (1) uncooperative, (2) mean, (3) inebriated, (4) threatening, (5) "crazy," (6) sexually inappropriate, and (7) aesthetically unappealing/overly appealing. They also identified 3 main characteristics of good clients: (1) communicative, (2) responsive, and (3) vulnerable. In addition, HIV test counselors used 4 strategies to deal with difficult clients: (1) received help from other counselors, (2) refused to test or threatened to refuse to test, (3) verbally confronted clients, and (4) "followed the forms" (ie, asked the necessary questions on the standard risk assessment forms). Results highlight the combined importance of patient characteristics, HIV test counselor characteristics, and the testing environment in contributing to difficult and good encounters in alternative HIV testing sites and point to the need for better training and support services in this area. PMID- 18400686 TI - Alcohol consumption and prehypertension: an investigation of university youth. AB - Prehypertension and heavy alcohol consumption increase the risk for primary hypertension (PH), a major predictor of cardiovascular-related morbidity and mortality. Although undergraduate college students have exhibited prehypertensive blood pressure (BP) levels and more than 40% of undergraduates drink heavily, few researchers have examined both risk factors in the university context. In this study, the authors collected BP and self-reported quantity and frequency of alcohol consumption data from 211 undergraduates (95 women). Logistic regression analyses showed that prehypertensive undergraduates (ie, those with systolic BP > or = 120 mm Hg or diastolic BP > or = 85 mm Hg) were nearly 4 times more likely to consume alcohol levels associated with increased risk for developing PH. Additional research on alcohol and PH among adolescents and undergraduates is needed, with particular reference to mechanisms and reducing the risk for morbidity and mortality emanating from cardiovascular disease. PMID- 18400687 TI - Memory for fatigue in chronic fatigue syndrome: relationships to fatigue variability, catastrophizing, and negative affect. AB - Fatigue in chronic fatigue syndrome (CFS) is usually assessed with retrospective measures rather than real-time momentary symptom assessments. In this study, the authors hypothesized that in participants with CFS, discrepancies between recalled and momentary fatigue would be related to catastrophizing, anxiety, and depression and to variability of momentary fatigue. They also expected that catastrophizing, anxiety, and depression would be associated with momentary fatigue. The authors asked 53 adults with CFS to carry electronic diaries for 3 weeks and record their experiences of momentary fatigue. The authors assessed participants' fatigue recall with weekly ratings and administered questionnaires for catastrophizing, depression, and anxiety. Recall discrepancy was significantly related to the variability of momentary fatigue. In addition, catastrophizing, depression, and momentary fatigue were all significantly related to recall discrepancy. Catastrophizing, depression, anxiety, and momentary negative affect were all significantly associated with momentary fatigue. The findings suggest that momentary fatigue in patients with CFS is related to modifiable psychological factors. PMID- 18400688 TI - What you don't know can't hurt you. . . . PMID- 18400689 TI - Aging of the immune system as a prognostic factor for human longevity. AB - Accumulating data are documenting an inverse relationship between immune status, response to vaccination, health, and longevity, suggesting that the immune system becomes less effective with advancing age and that this is clinically relevant. The mechanisms and consequences of age-associated immune alterations, designated immunosenescence, are briefly reviewed here. PMID- 18400690 TI - Neural systems controlling the drive to eat: mind versus metabolism. AB - With the bleak outlook that 75% of Americans will be overweight or obese in 10 years, it is essential to find efficient help very soon. Knowledge of the powerful and complex neural systems conferring the basic drive to eat is a prerequisite for designing efficient therapies. Recent studies suggest that the cross talk between brain areas involved in cognitive, emotional, and metabolic regulatory functions may explain why energy homeostasis breaks down for many predisposed individuals in our modern environment. PMID- 18400691 TI - Mitochondria: the hub of cellular Ca2+ signaling. AB - Mitochondria couple cellular metabolic state with Ca(2+) transport processes. They therefore control not only their own intra-organelle [Ca(2+)], but they also influence the entire cellular network of cellular Ca(2+) signaling, including the endoplasmic reticulum, the plasma membrane, and the nucleus. Through the detailed study of mitochondrial roles in Ca(2+) signaling, a remarkable picture of inter organelle communication has emerged. We here review the ways in which this system provides integrity and flexibility for the cell to cope with the countless demands throughout its life cycle and discuss briefly the mechanisms through which it can also drive cell death. PMID- 18400692 TI - Apical transporters for neutral amino acids: physiology and pathophysiology. AB - Absorption of amino acids in kidney and intestine involves a variety of transporters for different groups of amino acids. This is illustrated by inherited disorders of amino acid absorption, such as Hartnup disorder, cystinuria, iminoglycinuria, dicarboxylic aminoaciduria, and lysinuric protein intolerance, affecting separate groups of amino acids. Recent advances in the molecular identification of apical neutral amino acid transporters has shed a light on the molecular basis of Hartnup disorder and iminoglycinuria. PMID- 18400693 TI - The solute carrier 26 family of proteins in epithelial ion transport. AB - Transepithelial Cl(-) and HCO(3)(-) transport is critically important for the function of all epithelia and, when altered or ablated, leads to a number of diseases, including cystic fibrosis, congenital chloride diarrhea, deafness, and hypotension (78, 111, 119, 126). HCO(3)(-) is the biological buffer that maintains acid-base balance, thereby preventing metabolic and respiratory acidosis (48). HCO(3)(-) also buffers the pH of the mucosal layers that line all epithelia, protecting them from injury (2). Being a chaotropic ion, HCO(3)(-) is essential for solubilization of ions and macromolecules such as mucins and digestive enzymes in secreted fluids. Most epithelia have a Cl(-)/HCO(3) exchange activity in the luminal membrane. The molecular nature of this activity remained a mystery for many years until the discovery of SLC26A3 and the realization that it is a member of a new family of Cl(-) and HCO(3)(-) transporters, the SLC26 family (73, 78). This review will highlight structural features, the functional diversity, and several regulatory aspects of the SLC26 transporters. PMID- 18400696 TI - Prevalence and predictors of mental disorders among women in Sanliurfa, Southeastern Turkey. AB - BACKGROUND: Mental health is one of the most important public health issues because of major contributor to the global burden of disease. In this study, we examined the prevalence and predictors of mental disorders among married women from 15 to 49 years of age and the need for mental health services in the primary health care settings. METHODS: In this cross-sectional study, 270 women were selected using probability cluster sampling method at 95% confidence interval (91.5% response rate). The Structured Clinical Interview for DSM-IV (SCID-I) and women socio-demographic information form were used to collect data. RESULTS: Although the prevalence of mental disorder was 25.9% (8.5% with one diagnosis; 17.4% were two or more diagnoses), 4.7% of these women had contacted a carer in the last year for psychological reasons. According to the SCID-I assessment, the most prevalent diagnoses were major depressive disorder (7.3%), phobic disorder (4.8%) and posttraumatic stress disorder (3.6%). In this study, comorbid diagnoses were present in 67.2% of patients. Logistic regression analyses revealed that domestic violence, history of previous trauma, anemia and cutaneous leishmaniasis were significant predictors of any mental disorders (P < 0.05). CONCLUSIONS: These findings highlight the need for systematic development of community-based mental health services in conjunction with primary health care services for the screening, early identification and treatment of women suffering from mental disorders, and the improvement of anemia and cutaneous leishmaniasis control programme. PMID- 18400694 TI - Ion transport and energetics during cell death and protection. AB - During ischemia, ATP and phosphocreatine (PCr) decline, whereas intracellular hydrogen ion, intracellular sodium (Na(+)), calcium (Ca(2+)), and magnesium (Mg(2+)) concentrations all rise. If the ischemia is relatively short and there is little irreversible injury (cell death), PCr, pH, Na(+), Mg(2+), and Ca(2+) all recovery quickly on reperfusion. ATP recovery can take up to 24 h because of loss of adenine base from the cell and the need for de novo synthesis. There are correlative data showing that a sustained rise in Ca(2+) during ischemia and/or lack of recovery during reperfusion is associated with irreversible cell injury. Interventions that reduce the rise in Ca(2+) during ischemia and reperfusion have been shown to reduce cell death. Therefore, a better understanding of the mechanisms responsible for the rise in Ca(2+) during ischemia and early reperfusion could have important therapeutic implications. This review will discuss mechanisms involved in alterations in ions and high energy phosphate metabolites in perfused or intact heart during ischemia and reperfusion. PMID- 18400697 TI - Measuring burden of disease in two inner London boroughs using Disability Adjusted Life Years. AB - BACKGROUND: This paper uses the Disability Adjusted Life Year (DALY) to estimate disease burden at a local level and relates this to programme budget (PB) data. METHODS: We estimated DALY using the global burden of disease (GBD) template. For years of life lost, local mortality data were used and for years of life with disability, the GBD estimates from World Health Organization EURO A region (including the UK) were used. We used PB data to analyse how healthcare expenditure matched disease burden. RESULTS: In 2005 the burden of disease in Lambeth was estimated at 36,368 DALYs (13,515 DALYs lost per 100,000) and in Southwark was 34,196 DALYs (13,244 DALYs lost per 100,000). There were gender and area differences. The ranking is different when mortality and morbidity are combined compared with mortality alone. We estimated that the average spend per DALY lost in 2005 was 11,066 pounds in Lambeth and 9390 pounds in Southwark. CONCLUSIONS: We used a pragmatic approach to estimate overall disease burden providing a local, more comprehensive picture with important differences in spend by disease and health authority area. However, a more detailed approach to support decisions about prioritization based on modelling interventions that impact on avoidable burden of disease is recommended. PMID- 18400698 TI - Are alkylresorcinols accurate biomarkers for whole grain intake? PMID- 18400699 TI - Egg consumption in relation to cardiovascular disease and mortality: the story gets more complex. PMID- 18400700 TI - Consequences of smoking for body weight, body fat distribution, and insulin resistance. AB - Our aim was to critically evaluate the relations among smoking, body weight, body fat distribution, and insulin resistance as reported in the literature. In the short term, nicotine increases energy expenditure and could reduce appetite, which may explain why smokers tend to have lower body weight than do nonsmokers and why smoking cessation is frequently followed by weight gain. In contrast, heavy smokers tend to have greater body weight than do light smokers or nonsmokers, which likely reflects a clustering of risky behaviors (eg, low degree of physical activity, poor diet, and smoking) that is conducive to weight gain. Other factors, such as weight cycling, could also be involved. In addition, smoking increases insulin resistance and is associated with central fat accumulation. As a result, smoking increases the risk of metabolic syndrome and diabetes, and these factors increase risk of cardiovascular disease. In the context of the worldwide obesity epidemic and a high prevalence of smoking, the greater risk of (central) obesity and insulin resistance among smokers is a matter of major concern. PMID- 18400701 TI - Dietary energy density and diet variety as predictors of outcome in anorexia nervosa. AB - BACKGROUND: Anorexia nervosa (AN) is a serious psychiatric illness associated with significant morbidity and mortality. Successful treatment results in weight restoration, but recidivism is common, and the rate of relapse is estimated to be as high as 50%. Maintenance of a healthy diet is central to the recovery process, but the relation between diet and relapse has not been investigated in AN patients. OBJECTIVE: The objective of the study was to determine whether diet energy density and diet variety in recently weight-restored women with AN predict outcome. DESIGN: After gaining weight to a body mass index (BMI; in kg/m(2)) of > or = 20, 47 hospitalized women completed 4-d food records, from which a mean diet energy density score (DEDS) and a mean diet variety score (DVS) were calculated. Outcome was determined at study end by using modified Morgan-Russell criteria, and it was dichotomized as "treatment success" or "treatment failure." Data were analyzed by using Student's t test. A logistic regression model was constructed to evaluate the effects of DEDS, DVS, and caloric intake on outcome. RESULTS: Groups did not differ significantly in mean measures of age, admission and weight restored BMI, or caloric intake. However, DEDS and DVS were significantly higher in the success group than in the failure group. The success and failure groups were followed for a mean of 240 and 170 d, respectively. In the logistic regression model, DEDS (P = 0.016) and DVS (P = 0.048) but not caloric intake (P = 0.585) significantly predicted outcome. CONCLUSION: In recently weight-restored women with AN, lower DEDS and DVS but not caloric intake were associated with poor outcome. PMID- 18400702 TI - Parallel activation of de novo lipogenesis and stearoyl-CoA desaturase activity after 3 d of high-carbohydrate feeding. AB - BACKGROUND: High-carbohydrate (HC) diets increase de novo lipogenesis (DNL), but effects on stearoyl-CoA desaturase (SCD) are not so well studied. OBJECTIVE: The objective was to investigate DNL and SCD in liver and adipose tissue by using fatty acid ratios after short-term dietary intervention. DESIGN: Eight subjects consumed isoenergetic 3-d HC (10% fat; 75% carbohydrates) or higher fat (HF; 40% fat; 45% carbohydrates) diets (sugar to starch ratio: 60:40 for both) in a crossover study. Blood was taken from an artery and a vein draining subcutaneous adipose tissue. DNL and SCD activity were investigated by using the ratios of 16:0 to 18:2n-6 and of 16:1n-7 to 16:0, respectively. A test meal, including [U (13)C]palmitate was given to trace dietary fatty acid incorporation into VLDL triacylglycerol (TG). The conversion of intravenously infused [(2)H(2)]palmitic acid to [(2)H(2)]palmitoleic acid in VLDL-TG was quantified as a specific marker of hepatic SCD activity. RESULTS: The VLDL-TG 16:0/18:2n-6 ratio, which reflects hepatic DNL, was greater after the HC diet than after the HF diet (P = 0.02). With the HC diet, increased plasma TG concentrations correlated with 16:0/18:2n-6 ratios (r = 0.76, P = 0.028). Plasma VLDL-TG and adipose venous nonesterified fatty acid (NEFA) 16:1n-7/16:0 ratios were higher after the HC diet (fasting: P = 0.01 and P = 0.05, respectively; postprandial: P = 0.03 and P = 0.05, respectively). Changes in fasting VLDL-TG 16:0/18:2n-6 and 16:1n-7/16:0 ratios were associated (P = 0.06). The contribution of total fatty acids from splanchnic sources (including DNL) was higher after the HC diet (P = 0.02). Expression of lipogenic genes in subcutaneous adipose tissue was not significantly affected by diet. CONCLUSION: Parallel activation of DNL and SCD was found after a short period of HC feeding. PMID- 18400703 TI - Reduced oxidation of dietary fat after a short term high-carbohydrate diet. AB - BACKGROUND: Short-term high-carbohydrate (HC) diets induce metabolic alterations, including hypertriacylglycerolemia, in both the fasting and postprandial states. The underlying tissue-specific alterations in fatty acid metabolism are not well understood. OBJECTIVE: We investigated alterations in exogenous and endogenous fatty acid metabolism by using stable isotope tracers to label meal triacylglycerol and plasma fatty acids. DESIGN: Eight healthy subjects consumed isocaloric diets containing a high percentage of energy from carbohydrates or a higher percentage of energy from fat for 3 d in a randomized crossover dietary intervention study. A test meal containing [U-13C] palmitate was combined with intravenous infusion of [2H2] palmitate to label plasma fatty acids and VLDL triacylglycerol. Blood and breath samples were taken before the meal and for 6 h postprandially. Blood samples were drawn from the femoral artery and from veins draining subcutaneous adipose tissue and forearm muscle for monitoring of tissue specific metabolic substrate partitioning. RESULTS: Systemic triacylglycerol concentrations were increased in both fasting (P = 0.02) and postprandial (P = 0.02) periods, and a greater amount of infused labeled fatty acid appeared in VLDL triacylglycerol after the HC diet than after the higher-fat diet (P = 0.05). Significantly less 13CO2 was exhaled after the HC diet (P = 0.04) and significantly less production of 13CO2 was seen across forearm muscle (P = 0.04). Systemic 3-hydroxybutyrate was significantly lower, postprandially, after the HC diet (P = 0.02). CONCLUSION: Metabolic alterations suggestive of repartitioning of fatty acids away from oxidation toward esterification in both liver and muscle occur in response to short-term adaptation to a HC diet. PMID- 18400704 TI - Alkylresorcinols as biomarkers of whole-grain wheat and rye intake: plasma concentration and intake estimated from dietary records. AB - BACKGROUND: Alkylresorcinols (ARs), phenolic lipids exclusively present in the outer parts of wheat and rye grains, have been proposed as specific dietary biomarkers of whole-grain wheat and rye intake. OBJECTIVE: The objective was to validate plasma ARs as a biomarker of whole-grain wheat and rye intakes by studying the correlation between their plasma concentration and intake calculated from food records. DESIGN: In a randomized crossover study, 22 women and 8 men were given a defined amount of either whole-grain or refined-cereal-grain products to be included in their habitual diets for two 6-wk periods. Blood samples were collected and food intakes were recorded before and after each intervention period. RESULTS: Plasma AR concentrations were significantly higher after the whole-grain diet period than after the refined-grain period (P < 0.0001) and were well correlated with average daily AR intake estimated by self reported weighed food records (Spearman's r = 0.58, P < 0.001). CONCLUSION: Plasma AR concentrations are correlated with intake assessed by food records, which suggests that ARs are selective nutritional biomarkers for the intake of whole-grain wheat and rye. PMID- 18400705 TI - Predictors of iron status in well-nourished 4-y-old children. AB - BACKGROUND: Iron status in childhood is influenced by diet. Other factors affecting iron status at that age are unclear. OBJECTIVES: The objectives of the study were to evaluate iron status in 4-y-old children, to track that status from infancy to childhood, and to examine the associations of iron status with dietary factors, growth, and heredity. DESIGN: This study consisted of a longitudinal follow-up at age 4 y of children (n = 127) from the cohort of a study that began at age 6 mo. Blood samples and anthropometry were assessed in both children and their parents; food records were collected from children only. RESULTS: Dietary intake was not significantly correlated with hemoglobin concentrations, whereas the consumption of meat products had a positive effect on serum ferritin concentrations and mean corpuscular volume in boys (P = 0.015 and 0.04, respectively). The prevalences of anemia and iron deficiency were low, affecting 2 (1.8%) and 3 (2.8%) children, respectively; no child had iron deficiency anemia. There was significant within-subject tracking of hemoglobin and mean corpuscular volume from age 6 mo to 4 y. The mother's but not the father's hemoglobin correlated with the child's hemoglobin over time. CONCLUSIONS: Food choices had little effect on iron status. Hemoglobin concentrations and mean corpuscular volume were tracked from infancy to childhood. In healthy, well nourished children with a low prevalence of iron deficiency, the mother's hemoglobin was significantly associated with that of her child, but the underlying mechanism is unclear. PMID- 18400706 TI - Energy-dense, low-fiber, high-fat dietary pattern is associated with increased fatness in childhood. AB - BACKGROUND: Evidence for the dietary determinants of obesity in children is limited. OBJECTIVE: The objective was to identify a dietary pattern that explained dietary energy density (DED), fiber density (FD), and percentage of energy intake from fat and analyze its relation to fatness in children. DESIGN: The subjects were 521 (at ages 5 and 9 y) and 682 (at ages 7 and 9 y) children participating in the Avon Longitudinal Study of Parents and Children. Diet was assessed with the use of 3-d diet diaries at ages 5 and 7 y. Reduced rank regression derived a dietary pattern with the use of DED, fiber, and fat intake as intermediate variables. Fat mass was measured at age 9 y with the use of dual energy X-ray absorptiometry. Fat mass index (FMI) was calculated, and excess adiposity was defined (as the top quintile of logFMI). RESULTS: Pattern score at ages 5 and 7 y was correlated with DED (r = 0.8), FD (r = -0.7), and percentage of energy intake from fat (r = 0.5). An increase of 1 SD of pattern score at ages 5 and 7 y, respectively, was associated with a 0.15-kg (95% CI: -0.1, 0.45 kg) and a 0.28-kg (95% CI: 0.05, 0.53 kg) higher fat mass at age 9 y, after controlling for confounders. The adjusted odds of excess adiposity at age 9 y for children in quintile 5 compared with quintile 1 of dietary pattern score at ages 5 and 7 y, respectively, were 2.52 (95% CI: 1.13, 6.08) and 4.18 (95% CI: 2.07, 9.38). CONCLUSION: An energy-dense, low-fiber, high-fat diet is associated with higher fat mass and greater odds of excess adiposity in childhood. PMID- 18400707 TI - Comparison of the effects on insulin resistance and glucose tolerance of 6-mo high-monounsaturated-fat, low-fat, and control diets. AB - BACKGROUND: The effect of dietary fat and carbohydrate on glucose metabolism has been debated for decades. OBJECTIVE: The objective was to compare the effect of 3 ad libitum diets, different in type and amount of fat and carbohydrate, on insulin resistance and glucose tolerance subsequent to weight loss. DESIGN: Forty six nondiabetic, obese [mean (+/-SEM) body mass index (in kg/m(2)): 31.2 +/- 0.3] men (n = 20) and premenopausal women (n = 26) aged 28.0 +/- 0.7 y were randomly assigned to 1 of 3 diets after > or = 8% weight loss: 1) MUFA diet (n = 16): moderate in fat (35-45% of energy) and high in monounsaturated fatty acids ( > 20% of energy); 2) LF diet (n = 18): low-fat diet (20-30% of energy), and 3) control diet (n = 12): 35% of energy as fat ( > 15% of energy as saturated fatty acids). Protein accounted for 15% of energy in all 3 diets. A 2-h oral-glucose tolerance test (OGTT) was performed before and after the 6-mo dietary intervention. All foods were provided by a purpose-built supermarket. RESULTS: After 6 mo, the MUFA diet reduced fasting glucose (-3.0%), insulin (-9.4%), and the homeostasis model assessment of insulin resistance score (-12.1%). Compared with the MUFA diet, the control diet increased these variables [1.4% (P = 0.014), 21.2% (P = 0.030), and 22.8% (P = 0.015), respectively], as did the LF diet [1.4% (P = 0.090), 13.1% (P = 0.078), and 15.5% (P = 0.095), respectively]. No significant group differences were detected in glucose or insulin concentrations during the OGTT, in the Matsudas index, in body weight, or in body composition. CONCLUSION: A diet high in monounsaturated fat has a more favorable effect on glucose homeostasis than does the typical Western diet in the short term and may also be more beneficial than the official recommended low-fat diet during a period of weight regain subsequent to weight loss. PMID- 18400708 TI - Further characterization of a furanocoumarin-free grapefruit juice on drug disposition: studies with cyclosporine. AB - BACKGROUND: We previously established furanocoumarins as mediators of the interaction between grapefruit juice (GFJ) and the model CYP3A4 substrate felodipine in healthy volunteers using a GFJ devoid of furanocoumarins. It remains unclear whether furanocoumarins mediate drug-GFJ interactions involving CYP3A4 substrates that are also P-glycoprotein substrates. OBJECTIVE: The effects of furanocoumarin-free GFJ on drug disposition were further characterized by using the dual CYP3A4/P-glycoprotein substrate cyclosporine. DESIGN: By randomized crossover design, 18 healthy volunteers received cyclosporine (5 mg/kg) with 240 mL orange juice (control), GFJ, or furanocoumarin-free GFJ. Blood was collected over 24 h. Juice treatments were separated by > or = 1 wk. The effects of diluted extracts of each juice and of purified furanocoumarins on [3H]cyclosporine translocation in Caco-2 cells were then compared. RESULTS: The median (range) dose-corrected cyclosporine area under the curve and the maximum concentration with GFJ (P < or = 0.007), but not with furanocoumarin-free GFJ (P > or = 0.50), were significantly higher than those with orange juice [15.6 (6.7 33.5) compared with 11.3 (4.8-22.0) x 10(-3) h/L and 3.0 (1.6-5.8) compared with 2.4 (1.1-3.1) mL(-1), respectively]. The median time to reach maximum concentration and terminal elimination half-life were not significantly different between the juices (2-3 and 7-8 h, respectively; P > or = 0.08). Relative to vehicle, the GFJ extract, orange juice extract, and purified furanocoumarins partially increased apical-to-basolateral and decreased basolateral-to-apical [3H]cyclosporine translocation in Caco-2 cells, whereas the furanocoumarin-free GFJ extract had negligible effects. Reanalysis of the clinical juices identified polymethoxyflavones as candidate P-glycoprotein inhibitors in orange juice but not in GFJ. CONCLUSIONS: Furanocoumarins mediate, at least partially, the cyclosporine-GFJ interaction in vivo. A plausible mechanism involves the combined inhibition of enteric CYP3A4 and P-glycoprotein. PMID- 18400709 TI - A double-blind, placebo-controlled, randomized trial of the effects of dark chocolate and cocoa on variables associated with neuropsychological functioning and cardiovascular health: clinical findings from a sample of healthy, cognitively intact older adults. AB - BACKGROUND: In recent years, there has been increased interest in the potential health-related benefits of antioxidant- and phytochemical-rich dark chocolate and cocoa. OBJECTIVE: The objective of the study was to examine the short-term (6 wk) effects of dark chocolate and cocoa on variables associated with neuropsychological functioning and cardiovascular health in healthy older adults. DESIGN: A double-blind, placebo-controlled, fixed-dose, parallel-group clinical trial was used. Participants (n = 101) were randomly assigned to receive a 37-g dark chocolate bar and 8 ounces (237 mL) of an artificially sweetened cocoa beverage or similar placebo products each day for 6 wk. RESULTS: No significant group (dark chocolate and cocoa or placebo)-by-trial (baseline, midpoint, and end of-treatment assessments) interactions were found for the neuropsychological, hematological, or blood pressure variables examined. In contrast, the midpoint and end-of-treatment mean pulse rate assessments in the dark chocolate and cocoa group were significantly higher than those at baseline and significantly higher than the midpoint and end-of-treatment rates in the control group. Results of a follow-up questionnaire item on the treatment products that participants believed they had consumed during the trial showed that more than half of the participants in both groups correctly identified the products that they had ingested during the experiment. CONCLUSIONS: This investigation failed to support the predicted beneficial effects of short-term dark chocolate and cocoa consumption on any of the neuropsychological or cardiovascular health-related variables included in this research. Consumption of dark chocolate and cocoa was, however, associated with significantly higher pulse rates at 3- and 6-wk treatment assessments. PMID- 18400710 TI - Iron absorption in young Indian women: the interaction of iron status with the influence of tea and ascorbic acid. AB - BACKGROUND: Ascorbic acid (AA) enhances and tea inhibits iron absorption. It is unclear whether iron status influences the magnitude of this effect. OBJECTIVE: We evaluated the influence of the iron status of young women on iron absorption from a rice meal with or without added tea or AA. DESIGN: Two stable-isotope iron absorption studies were made in 2 groups of 10 subjects with iron deficiency anemia (IDA) and 10 subjects who were iron replete (control subjects). In study 1, the reference rice meal was fed alone or with 1 or 2 cups of black tea. In study 2, the reference meal was fed alone or with AA (molar ratio to iron, 2:1 or 4:1). Iron absorption was measured by the erythrocyte incorporation of (57)Fe and (58)Fe labels at 14 d. RESULTS: Mean fractional iron absorption from the reference rice meal was approximately 2.5 times as great in the IDA group as in the control group (P < 0.05). The consumption of 1 or 2 cups of tea decreased iron absorption in the control subjects by 49% (P < 0.05) or 66% (P < 0.01), respectively, and in the IDA group by 59% or 67% (P < 0.001 for both), respectively. AA (molar ratio to iron, 2:1 or 4:1) increased iron absorption by 270% or 343%, respectively, in control subjects and by 291% or 350%, respectively, in subjects with IDA (P < 0.001). CONCLUSIONS: The inhibitory effect of tea and the enhancing effect of AA on iron absorption were similar in the 2 groups. Overall differences in iron absorption in the 2 groups, however, continued to be dictated by iron status. PMID- 18400711 TI - Maternal n-3, n-6, and trans fatty acid profile early in pregnancy and term birth weight: a prospective cohort study. AB - BACKGROUND: Maternal n-3, n-6, and trans fatty acids are claimed to affect fetal growth, yet evidence is limited. OBJECTIVE: We investigated the association between maternal n-3, n-6, and trans fatty acids measured early in pregnancy and fetal growth. DESIGN: Amsterdam pregnant women (n = 12 373) were invited to complete a questionnaire (response 67%) and donate blood around the 12th pregnancy week for nutrient analysis. For 4336 women, fatty acid concentrations were measured in plasma phospholipids (gas-liquid chromatography). Associations of these concentrations with birth weight and small-for-gestational-age (SGA) risk were analyzed (liveborn singleton term deliveries, n = 3704). RESULTS: Low concentrations of individual n-3 fatty acids and 20:3n-6, the precursor of arachidonic acid (20:4n-6), but high concentrations of the other n-6 fatty acids and the main dietary trans fatty acid (18:1n-9t) were associated with lower birth weight (estimated difference in univariate analysis -52 to -172 g for extreme quintile compared with middle quintile). In general, SGA risk increased accordingly. After adjustment for physiologic, lifestyle-related and sociodemographic factors, low concentrations of most n-3 fatty acids and 20:3n-6 and high concentrations of 20:4n-6 remained associated with lower birth weight ( 52 to -57 g), higher SGA risk, or both (odds ratios: 1.38-1.50). Infants of the 7% of women with the most adverse fatty acid profile were on average 125 g lighter and twice as likely to be small for gestational age. CONCLUSION: An adverse maternal fatty acid profile early in pregnancy is associated with reduced fetal growth, which, if confirmed, gives perspective for the dietary prevention of lower birth weight. PMID- 18400712 TI - Choline concentrations in human maternal and cord blood and intelligence at 5 y of age. AB - BACKGROUND: Animal studies indicate that maternal prenatal choline supplementation leads to permanent enhancement of attention and spatial memory abilities in offspring, whereas dietary choline restriction during pregnancy impairs cognitive function in offspring. The association between gestational choline concentrations and neurodevelopmental outcome in humans has not been studied. OBJECTIVE: Our objective was to assess the relation between maternal and cord blood choline concentrations and child intelligence quotient (IQ) scores at 5 y of age. DESIGN: With data and samples from a prospective study (n = 404 maternal-child pairs), serum concentrations of free and total choline were measured in maternal serum at 4 gestational age intervals (16-18 wk, 24-26 wk, 30 32 wk, and 36-38 wk) and in cord blood. Child IQ at 5 y of age was assessed with the Wechsler Preschool and Primary Scale of Intelligence-Revised. Multiple regression techniques were used to estimate the relation between choline concentrations and Full Scale IQ, Verbal and Performance IQ, and subscales that assess spatial relation and memory ability while adjusting for other factors that affect IQ. RESULTS: There was no effect at gestational ages 16-18 wk, 24-26 wk, 30-32 wk, and 36-38 wk or in cord blood of serum concentrations of free or total choline on Full Scale child IQ or on selected scales related to visuospatial processing and memory. CONCLUSION: Gestational and newborn choline concentrations in the physiologic range showed no correlation with childhood intelligence. PMID- 18400713 TI - Genetic and shared environmental influences on children's 24-h food and beverage intake: sex differences at age 7 y. AB - BACKGROUND: The genetics of habitual food and beverage intake in early childhood is poorly understood. OBJECTIVE: The objective was to test the magnitude of genetic and environmental influences on 24-h food and beverage intake in 7-y-old children. The association between intake of specific food-beverage categories and child body mass index (BMI; in kg/m(2)) was also tested. DESIGN: A classic twin design was conducted, using the MacArthur Longitudinal Study of Twins. There were 792 children, including 396 boys from 102 monozygotic and 96 dizygotic twin pairs and 396 girls from 112 monozygotic and 86 dizygotic twin pairs; Children's 24-h dietary intake was estimated by parental recall, from which 9 composite food beverage categories were derived. Height and weight were converted to BMI. Biometrical analyses of children's daily intake of food-beverage categories and BMI were conducted. RESULTS: There was consistent evidence of genetic influences on children's 24-h intake of food and beverages (servings/d), especially among boys. Seven categories showed significant heritability estimates among boys, ranging from 12% (fish and lemon) to 79% (peanut butter and jelly). Only 3 categories showed significant heritability estimates among girls, ranging from 20% (bread and butter) to 56% (fish and lemon). BMI showed a genetic correlation only with bread and butter intake in girls. CONCLUSION: The magnitude of genetic and environmental influences on children's 24-h food and beverage intake differed for boys and girls, which suggests sex differences in the development of eating patterns. Heritability estimates were generally large, although other eating phenotypes may be necessary for identifying genetic correlations with adiposity. PMID- 18400714 TI - Effect of long-chain polyunsaturated fatty acid supplementation of preterm infants on disease risk and neurodevelopment: a systematic review of randomized controlled trials. AB - BACKGROUND: Supplementation of preterm formulas with long-chain polyunsaturated fatty acids (LCPUFAs) is based on their effectiveness to increase blood status and improve visual outcomes. Dispute remains over their efficacy on global development. OBJECTIVE: The objective was to compare the effects of LCPUFA supplemented with those of control formulas on neurodevelopment and diseases associated with prematurity. DESIGN: We systematically reviewed randomized controlled trials involving preterm infants that tested LCPUFA-supplemented formulas. The weighted mean differences (WMDs) in neurodevelopmental scores and relative risk (RR) of disease were calculated to compare infants fed LCPUFA supplemented formula with those fed control formula. RESULTS: No clear differences in Bayley Scales of Infant Development (BSID) scores were observed between groups. Mental development of LCPUFA-supplemented infants was 3.4 points higher than that of control infants with BSID version II (WMD: 3.44; 95% CI: 0.56, 6.31; P = 0.02; n = 879), although it was driven by 2 trials with large effect sizes and wide CIs. Psychomotor development was lower in supplemented infants tested with BSID version I (WMD: -7.99; 95% CI: -14.00, -1.99; P = 0.009; n = 87); however, it was limited by small sample sizes. No differences in the RR of sepsis (1.08; 95% CI: 0.79, 1.46; P = 0.63; n = 1333) or in the RR of necrotizing enterocolitis (1.13; 95% CI: 0.62, 2.04; P = 0.69; n = 1333) were found. Similarly, the risks of retinopathy of prematurity, intraventricular hemorrhage, and bronchopulmonary dysplasia were not different between groups, but data were limited by small sample sizes and trials with an increased risk of bias. CONCLUSIONS: LCPUFA-supplemented formula does not alter the risk of NEC or sepsis. Further work is needed to determine the extent of benefit of LCPUFA supplemented formula on the mental development of preterm infants. PMID- 18400715 TI - Effects of alpha-lactalbumin-enriched formula containing different concentrations of glycomacropeptide on infant nutrition. AB - BACKGROUND: Formula-fed infants have growth and plasma amino acid patterns different from those of breastfed infants. OBJECTIVE: alpha-Lactalbumin is a major protein in human milk, and the addition of bovine alpha-lactalbumin to infant formula has been proposed to modify the plasma amino acid pattern of the recipient infant, possibly allowing a reduction in the protein content of the formula, which may affect growth. DESIGN: We compared breastfed infants and infants fed standard formula or alpha-lactalbumin-enriched formulas (25% of protein) with glycomacropeptide accounting for 15% or 10% of the protein. The protein content of each formula was 13.1 g/L. Ninety-six infants aged 6 +/- 2 wk were recruited. Anthropometric measures were recorded, and interviews were conducted at enrollment and monthly until 6 mo of age. Blood samples were collected at enrollment and at 4 and 6 mo. RESULTS: Formula intake did not differ between groups, and weight gain in the alpha-lactalbumin-enriched formula groups were similar to that of the breastfed infants. The standard formula group gained significantly more weight than did the breastfed infants. All formula-fed infants had significantly higher plasma concentrations of most essential amino acids at 4 and 6 mo than did the breastfed infants, and serum urea nitrogen was also higher in the formula-fed infants. Insulin and leptin concentrations did not differ between groups. CONCLUSIONS: Compared with standard formula-fed infants, infants fed formula with a modified protein composition had growth patterns more similar to those of breastfed infants. All formula-fed groups had plasma amino acid concentrations similar to or higher than those of breastfed infants. This indicates that the protein content of alpha-lactalbumin-enriched formula can be further reduced, which should be evaluated. PMID- 18400716 TI - Home fortification of complementary foods with micronutrient supplements is well accepted and has positive effects on infant iron status in Ghana. AB - BACKGROUND: Micronutrient deficiencies are common during infancy, and optimal approaches for their prevention need to be identified. OBJECTIVE: The objective was to compare the efficacy and acceptability of Sprinkles (SP), crushable Nutritabs (NT), and fat-based Nutributter (NB; 108 kcal/d), which provide 6, 16, and 19 vitamins and minerals, respectively, when used for home fortification of complementary foods. DESIGN: Ghanaian infants were randomly assigned to receive SP (n = 105), NT (n = 105), or NB (n = 103) daily from 6 to 12 mo of age. We assessed dietary intake, morbidity, and compliance weekly. Hemoglobin and plasma ferritin, TfR, C-reactive protein, and zinc were measured at 6 and 12 mo. We used an exit interview to assess acceptability. A randomly selected control group of infants who received no intervention (NI; n = 96) were assessed at 12 mo. RESULTS: All supplements were well accepted, and the mean percentage of days that supplements were consumed (87%) did not differ between groups. At 12 mo, all 3 intervention groups had significantly higher ferritin and lower TfR concentrations than did the NI control group. Mean (+/- SD) hemoglobin was significantly higher in NT (112 +/- 14 g/L) and NB (114 +/- 14 g/L) but not in SP (110 +/- 14 g/L) infants than in NI infants (106 +/- 14 g/L). The prevalence of iron deficiency anemia was 31% in the NI control group compared with 10% in the intervention groups combined (P < 0.0001). CONCLUSION: All 3 options for home fortification of complementary foods are effective for reducing the prevalence of iron deficiency in such populations. PMID- 18400717 TI - Comparison of cytokine modulation by natural peroxisome proliferator-activated receptor gamma ligands with synthetic ligands in intestinal-like Caco-2 cells and human dendritic cells--potential for dietary modulation of peroxisome proliferator-activated receptor gamma in intestinal inflammation. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPARgamma) plays a role in the regulation of intestinal inflammation and is activated by both natural (polyunsaturated fatty acid; PUFAs) and synthetic (troglitazone) ligands. The fatty acid content of defined formula diets may play a role in mediating the antiinflammatory effect, but the mechanism is unclear. OBJECTIVE: We evaluated to what extent the effect of PUFAs on intestinal inflammation is mediated via PPARgamma. DESIGN: The human enterocyte-like cell line Caco-2 and human dendritic cells were stimulated by interleukin (IL) 1beta and lipoprotein polysaccharide, respectively, in the presence of PPARgamma agonists (troglitazone or PUFAs) or antagonist (GW9662). Five PUFAs were tested: alpha-linolenic acid (ALA), conjugated linoleic acid (CLA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and gamma-linolenic acid (GLA). Cytokine production was measured by enzyme linked immunosorbent assay and PPARgamma, I-kappaB, and inducible nitric oxide synthase (iNOS) expression by Western blot. RESULTS: In Caco-2 cells, IL-6 secretion was significantly decreased by troglitazone, DHA, EPA, and GLA. IL-8 production was significantly decreased by troglitazone, ALA, DHA, EPA, and GLA. PPARgamma expression was significantly increased by troglitazone, DHA, and EPA. iNOS expression was significantly decreased by troglitazone, DHA, and EPA. Troglitazone and PUFAs at 0.1 mumol/L tended to increase the expression of I kappaB. Addition of GW9662 reversed the effect of troglitazone and PUFAs at 0.1 mumol/L on IL-8 production and decreased the expression of PPARgamma. EPA and DHA also modulated the dendritic cell response to lipoprotein polysaccharide. CONCLUSIONS: The tested PUFAs exerted an antiinflammatory effect in vitro in both models. This effect of PUFAs in Caco-2 cells is similar to that of troglitazone on intestinal inflammation mediated by PPARgamma, and the potency of the antiinflammatory effect is linked to the number of double bonds. PMID- 18400718 TI - Effect of n-3 polyunsaturated fatty acids on Barrett's epithelium in the human lower esophagus. AB - BACKGROUND: Epidemiologic studies suggest a reduced risk of esophageal adenocarcinoma in populations with a high consumption of fish, and n-3 fatty acids inhibit experimental carcinogenesis. One possible explanation is the suppression of eicosanoid production through inhibition of cyclooxygenase 2 (COX 2). OBJECTIVE: The objective was to determine the effects of dietary supplementation with the n-3 fatty acid eicosapentaenoic acid (EPA) on a number of biological endpoints in Barrett's esophagus. DESIGN: Fifty-two participants with known Barrett's esophagus underwent endoscopy. Biopsy samples were obtained from a recorded level within the area of Barrett's esophagus, and then 27 patients were randomly assigned to consume EPA capsules (1.5 g/d) for 6 mo or no supplement (controls). At the end of this period, patients again underwent endoscopy, and biopsy samples were collected at the same level. Tissue samples were analyzed for mucosal lipid, prostaglandin E2, leukotriene B4, COX-2 protein, and RNA concentrations. Cellular proliferation was also measured, by Ki-67 immunohistochemistry. RESULTS: The EPA content of esophageal mucosa increased over the study period in the n-3-supplemented subjects and was significantly different from the content in the controls (P < 0.01). There was also a significant decline in COX-2 protein concentrations (measured by immunoblotting) in the n-3 group, and the difference was significant from that in the controls (P < 0.05); no difference in COX-2 RNA concentrations was observed between groups. This change in COX-2 protein was inversely related to the change in EPA content (P < 0.05). There was no significant difference in the change in prostaglandin E2, leukotriene B4, or cellular proliferation between the 2 groups. CONCLUSION: Supplementation with EPA significantly changed n-3 fatty acid concentrations and reduced COX-2 concentrations in Barrett's tissue. PMID- 18400719 TI - Alcohol drinking frequency in relation to subsequent changes in waist circumference. AB - BACKGROUND: Cross-sectional studies have reported a lower prevalence of abdominal obese persons among frequent drinkers than among nonfrequent drinkers. OBJECTIVE: We tested the hypothesis that drinking frequency is associated with subsequent changes in waist circumference. DESIGN: Data come from a prospective cohort study conducted in 1993-1997 (baseline) and 1999-2002 (follow-up) and included 43 543 men and women. Baseline information on alcohol drinking frequency was related to 1) change in waist circumference by linear regression and 2) major gain and major loss in waist circumference (defined as waist change in the lowest or highest quintile of waist changes) by polytomous logistic regression, also taking into account amount of alcohol intake. RESULTS: Drinking frequency was inversely associated with changes in waist circumference in women and was unassociated with changes in waist circumference in men. Drinking frequency was unassociated with major waist loss but was inversely associated with major waist gain: odds ratios among men were 0.97 (95% CI: 0.73, 1.28), 0.95 (95% CI: 0.81, 1.12), 0.88 (95% CI: 0.77, 0.99), 0.82 (95% CI: 0.71, -0.95), and 0.79 (95% CI: 0.69, 0.9) for never drinking, drinking on 1, 2-4, 5-6, and 7 d/wk, respectively, compared with men who drank alcohol on <1 d/wk (P for trend < 0.0001). Results for women were similar. Adjustment for the amount of alcohol intake or total energy intake did not affect results considerably. CONCLUSIONS: Drinking pattern may be associated with development of abdominal obesity; in this prospective study, drinking frequency was inversely associated with major waist gain and was unassociated with major waist loss. PMID- 18400720 TI - Egg consumption in relation to cardiovascular disease and mortality: the Physicians' Health Study. AB - BACKGROUND: A reduction in dietary cholesterol is recommended to prevent cardiovascular disease (CVD). Although eggs are important sources of cholesterol and other nutrients, limited and inconsistent data are available on the effects of egg consumption on the risk of CVD and mortality. OBJECTIVE: We aimed to examine the association between egg consumption and the risk of CVD and mortality. DESIGN: In a prospective cohort study of 21,327 participants from Physicians' Health Study I, egg consumption was assessed with an abbreviated food questionnaire. Cox regression was used to estimate relative risks. RESULTS: In an average follow-up of 20 y, 1550 new myocardial infarctions (MIs), 1342 incident strokes, and 5169 deaths occurred. Egg consumption was not associated with incident MI or stroke in a multivariate Cox regression. In contrast, adjusted hazard ratios (95% CI) for mortality were 1.0 (reference), 0.94 (0.87, 1.02), 1.03 (0.95, 1.11), 1.05 (0.93, 1.19), and 1.23 (1.11, 1.36) for the consumption of <1, 1, 2-4, 5-6, and > or = 7 eggs/wk, respectively (P for trend < 0.0001). This association was stronger among diabetic subjects, in whom the risk of death in a comparison of the highest with the lowest category of egg consumption was twofold (hazard ratio: 2.01; 95% CI: 1.26, 3.20; P for interaction = 0.09). CONCLUSIONS: Infrequent egg consumption does not seem to influence the risk of CVD in male physicians. In addition, egg consumption was positively related to mortality, more strongly so in diabetic subjects, in the study population. PMID- 18400721 TI - Glycemic load, glycemic index, and pancreatic cancer risk in the Netherlands Cohort Study. AB - BACKGROUND: Recent studies of pancreatic cancer suggest a role for hyperinsulinemia in carcinogenesis. Because insulin is secreted in response to elevated blood glucose concentrations, dietary factors that increase these concentrations may be important in pancreatic carcinogenesis. OBJECTIVE: The objective was to examine prospectively the relation between pancreatic cancer risk and dietary glycemic load (GL), overall glycemic index (GI), and intake of total carbohydrates and mono- and disaccharides. DESIGN: The Netherlands Cohort Study consisted of 120,852 men and women who completed a baseline questionnaire in 1986. After 13.3 y of follow-up, 408 pancreatic cancer cases were detected, 66% of which were microscopically confirmed. A validated 150-item food-frequency questionnaire, completed at baseline, was used to calculate carbohydrate and mono and disaccharide intakes and the GL and GI of the diet. RESULTS: Dietary GL, GI, or intake of carbohydrates and mono- and disaccharides were not associated with pancreatic cancer risk in this cohort. Also, the associations were not modified by sex. Our results did not change after the analysis was restricted to microscopically confirmed pancreatic cancer cases or after individuals who reported a history of diabetes at baseline were excluded from the analyses. CONCLUSIONS: Overall, our findings do not support the hypothesis that GL, GI, or intake of carbohydrates and mono- and disaccharides are positively associated with pancreatic cancer risk. This is in agreement with previous prospective studies that investigated the relation between GL and GI and pancreatic cancer risk. PMID- 18400722 TI - Diet patterns and breast cancer risk in Hispanic and non-Hispanic white women: the Four-Corners Breast Cancer Study. AB - BACKGROUND: There is a lower incidence of breast cancer among Hispanic women than among non-Hispanic white women. Little is known about the role of diet in this difference. OBJECTIVE: We examined the associations of dietary patterns (Western, Prudent, Native Mexican, Mediterranean, and Dieter) with risk for breast cancer in Hispanic women (757 cases, 867 controls) and non-Hispanic white women (1524 cases, 1598 controls) from the Four-Corners Breast Cancer Study. DESIGN: Dietary intake, physical activity, and other exposures were assessed by using interviews. Dietary patterns were defined via factor analysis. Risk was assessed by using logistic regression with adjustment for age, center, education, smoking, total activity, calories, dietary fiber, dietary calcium, height, parity, recent hormone exposure, family history of breast cancer, menopausal status, and body mass index x recent hormone exposure. RESULTS: The Western (odds ratio for highest versus lowest quartile: 1.32; 95% CI: 1.04, 168; P for trend < 0.01) and Prudent (1.42; 1.14, 1.77; P for trend < 0.01) dietary patterns were associated with greater risk, and the Native Mexican (0.68; 0.55, 0.85; P for trend < 0.01) and Mediterranean (0.76; 0.63, 0.92; P for trend < 0.01) dietary patterns were associated with lower risk of breast cancer. Body mass index modified the associations of the Western diet and breast cancer among postmenopausal women and those of the Native Mexican diet among premenopausal women. CONCLUSIONS: Associations of dietary patterns with breast cancer risk varied by menopausal and body mass index status, but there was little difference in associations between non-Hispanic white and Hispanic women. PMID- 18400723 TI - Dietary intake of vitamin K and risk of prostate cancer in the Heidelberg cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC Heidelberg). AB - BACKGROUND: Anticarcinogenic activities of vitamin K have been observed in various cancer cell lines, including prostate cancer cells. Epidemiologic studies linking dietary intake of vitamin K with the development of prostate cancer have not yet been conducted. OBJECTIVE: We evaluated the association between dietary intake of phylloquinone (vitamin K1) and menaquinones (vitamin K2) and total and advanced prostate cancer in the Heidelberg cohort of the European Prospective Investigation into Cancer and Nutrition. DESIGN: At baseline, habitual dietary intake was assessed by means of a food-frequency questionnaire. Dietary intake of phylloquinone and menaquinones (MK-4-14) was estimated by using previously published HPLC-based food-content data. Multivariate-adjusted relative risks of total and advanced prostate cancer in relation to intakes of phylloquinone and menaquinones were calculated in 11 319 men by means of Cox proportional hazards regression. RESULTS: During a mean follow-up time of 8.6 y, 268 incident cases of prostate cancer, including 113 advanced cases, were identified. We observed a nonsignificant inverse association between total prostate cancer and total menaquinone intake [multivariate relative risk (highest compared with lowest quartile): 0.65; 95% CI: 0.39, 1.06]. The association was stronger for advanced prostate cancer (0.37; 0.16, 0.88; P for trend = 0.03). Menaquinones from dairy products had a stronger inverse association with advanced prostate cancer than did menaquinones from meat. Phylloquinone intake was unrelated to prostate cancer incidence (1.02; 0.70, 1.48). CONCLUSIONS: Our results suggest an inverse association between the intake of menaquinones, but not that of phylloquinone, and prostate cancer. Further studies of dietary vitamin K and prostate cancer are warranted. PMID- 18400724 TI - Inverse relation between dietary intake of naturally occurring plant sterols and serum cholesterol in northern Sweden. AB - BACKGROUND: Plant sterols are bioactive compounds, found in all vegetable foods, which inhibit cholesterol absorption. Little is known about the effect of habitual natural dietary intake of plant sterols. OBJECTIVE: We investigated the relation between plant sterol density (in mg/MJ) and serum concentrations of cholesterol in men and women in northern Sweden. DESIGN: The analysis included 37 150 men and 40 502 women aged 29-61 y, all participants in the Vasterbotten Intervention Program. RESULTS: Higher plant sterol density was associated with lower serum total cholesterol in both sexes and with lower LDL cholesterol in women. After adjustment for age, body mass index (in kg/m(2)), and (in women) menopausal status, men with high plant sterol density (quintile 5) had 0.15 mmol/L (2.6%) lower total serum cholesterol (P for trend = 0.001) and 0.13 mmol/L (3.1%) lower LDL cholesterol (P = 0.062) than did men with low plant sterol density (quintile 1). The corresponding figures for women were 0.20 mmol/L (3.5%) lower total serum cholesterol (P for trend < 0.001) and 0.13 mmol/L (3.2%) lower LDL cholesterol (P for trend = 0.001). CONCLUSIONS: The present study is the second epidemiologic study to show a significant inverse relation between naturally occurring dietary plant sterols and serum cholesterol. To the extent that the associations found truly mirror plant sterol intake and not merely a diet high in vegetable fat and fiber, it highlights the importance of considering the plant sterol content of foods both in primary prevention of cardiovascular disease and in the dietary advice incorporated into nutritional treatment of patients with hyperlipidemia. PMID- 18400725 TI - Consumption of cured meats and prospective risk of chronic obstructive pulmonary disease in women. AB - BACKGROUND: Most cured meats contain nitrites. Nitrites generate oxidative nitrative stress and were shown in animal models to cause emphysema. Prospective epidemiologic data on cured meats and chronic obstructive pulmonary disease (COPD), however, are sparse. OBJECTIVE: We examined the relation between cured meat consumption and the prospective risk of newly diagnosed COPD in women. DESIGN: This was a prospective cohort study of 71 531 women from the Nurses' Health Study who completed a validated dietary questionnaire at baseline in 1984 and had no baseline COPD or a report of asthma. Participants were aged 38-63 y in 1984 and were followed for 16 y. RESULTS: A total of 750 new cases of COPD were documented during the follow-up. Cured meat consumption was positively associated with COPD risk after adjustment for age, smoking, and multiple other potential confounders. The adjusted relative risks of COPD across categories of cured meat consumption (never or almost never, 1-3 servings/mo, 1 serving/wk, 2-3 servings/wk, and > or = 4 servings/wk) were 1.0, 1.14 (95% CI: 0.78, 1.66), 1.15 (95% CI: 0.79, 1.69), 1.40 (95% CI: 0.96, 2.05), and 1.51 (95% CI: 1.00, 2.27), respectively, (P for trend = 0.005). This positive association was present among both past (P for trend = 0.02) and current (P for trend = 0.03) smokers. No association was observed among never smokers, probably because of the small number of COPD cases in these women. CONCLUSION: Frequent cured meat consumption was associated with an increased risk of newly diagnosed COPD among women who smoke. PMID- 18400727 TI - Functional effects of a common single-nucleotide polymorphism (GPX4c718t) in the glutathione peroxidase 4 gene: interaction with sex. AB - BACKGROUND: Selenium is essential for health in humans. Selenium is present as selenocysteine in selenoproteins such as the glutathione peroxidases (GPx). Selenocysteine incorporation requires specific structures in the 3'untranslated region (3'UTR) of selenoprotein mRNAs. OBJECTIVE: This study investigated the functional significance of the single-nucleotide polymorphism (SNP) GPx4c718t within the 3'UTR of the GPx4 gene. DESIGN: A selenium supplementation trial was carried out with prospectively genotyped individuals of both homozygote genotypes for this SNP. Blood samples were analyzed at baseline, after a 6-wk supplementation with 100 mug Se as sodium selenite/d, and during a 6-wk washout period. RNA-protein binding studies were carried out in vitro. RESULTS: Both lymphocyte GPx1 protein concentrations and plasma GPx3 activity increased significantly after selenium supplementation in CC but not TT participants. After selenium withdrawal, there was a significant fall in both lymphocyte GPx4 protein concentrations and GPx4 activity in TT but not in CC participants; this effect was modulated by sex. RNA-protein binding assays showed that both T and C variants of transcripts corresponding to the GPx4 3'UTR formed complexes in vitro and that the C variant bound more strongly than did either the T variant or the GPx1 3'UTR. CONCLUSIONS: The GPX4c718t SNP both alters protein binding to the 3'UTR in vitro and influences the concentration of lymphocyte GPx4 and other selenoproteins in vivo. The latter is consistent with competition for selenium in selenoprotein synthesis, and, at low selenium intake, the SNP thus may influence susceptibility to disease. PMID- 18400726 TI - Selected antioxidants and risk of hormone receptor-defined invasive breast cancers among postmenopausal women in the Women's Health Initiative Observational Study. AB - BACKGROUND: Few studies have evaluated carotenoids and vitamins C and E in association with the risk of breast cancers defined by estrogen receptor (ER) and progesterone receptor (PR) status. OBJECTIVE: We examined the associations between dietary and supplemental intakes of these nutrients and risk of breast cancers jointly defined by both ER and PR status among postmenopausal women. DESIGN: Our investigation was conducted in the Women's Health Initiative Observational Study. After following 84 805 women for an average of 7.6 y, 2879 incident invasive breast cancer cases had been ascertained, of whom 2509 had receptor data. We used Cox proportional hazards models to assess the associations of interest. RESULTS: Dietary alpha-carotene (highest versus lowest quintile: RR = 0.83; 95% CL = 0.70, 0.99; P for trend = 0.019), beta-carotene (highest versus lowest quintile: RR = 0.78; 95% CL = 0.66, 0.94; P for trend = 0.021), and lycopene (highest versus lowest quintile: RR = 0.85; 95% CL = 0.73, 1.00; P for trend = 0.064) were inversely associated with risk of ER+PR+breast cancer, but not with other breast cancer groups jointly defined by ER and PR status. Total or supplemental beta-carotene and dietary intakes of lutein+zeaxanthin and beta cryptoxanthin were not associated with breast cancers defined by ER and PR status. Vitamin E (regardless of source) and dietary vitamin C were not associated with breast cancer. However, total and supplemental vitamin C intake had weak positive associations with breast cancer overall. CONCLUSION: Dietary intake of certain carotenoids might be differentially associated with risk of invasive breast cancers jointly defined by ER and PR status among postmenopausal women. PMID- 18400729 TI - Low vitamin D status adversely affects bone health parameters in adolescents. AB - BACKGROUND: The effects of subclinical vitamin D deficiency on bone mineral density (BMD) and bone turnover in adolescents, especially in boys, are unclear. OBJECTIVE: We aimed to investigate the relations of different stages of vitamin D status and BMD and bone turnover in a representative sample of adolescent boys and girls. DESIGN: BMD was measured by dual-energy X-ray absorptiometry at the nondominant forearm and dominant heel in a random sample of 12- (n = 260) and 15 y-old (n = 239) boys and 12- (n = 266) and 15-y-old (n = 250) girls. Serum 25 hydroxyvitamin D, parathyroid hormone, osteocalcin, and type I collagen cross linked C-telopeptide were assessed by using enzyme-linked immunoassays. Relations between vitamin D status and bone health indexes were assessed by using regression modeling. RESULTS: Using multivariate regression to adjust for potential physical, lifestyle, and dietary confounding factors, we observed that 12- and 15-y-old girls with high vitamin D status (>/=74.1 nmol/L) had significantly greater forearm (but not heel) BMD (beta = 0.018; SE = 0.008; P < 0.05 for each age group) and lower serum parathyroid hormone concentrations and bone turnover markers than did those with low vitamin D status. These associations were evident in subjects sampled throughout the year and in winter only. There was no significant relation between vitamin D status and BMD in boys. CONCLUSIONS: Maintaining serum 25-hydroxyvitamin D concentrations above approximately 50 nmol/L throughout the year may be a cost-effective means of improving bone health. Increased emphasis on exploring strategies for improving vitamin D status in adolescents is needed. PMID- 18400728 TI - Luminal fructose inhibits rat intestinal sodium-phosphate cotransporter gene expression and phosphate uptake. AB - BACKGROUND: While searching by microarray for sugar-responsive genes, we inadvertently discovered that sodium-phosphate cotransporter 2B (NaPi-2b) mRNA concentrations were much lower in fructose-perfused than in glucose-perfused intestines of neonatal rats. Changes in NaPi-2b mRNA abundance by sugars were accompanied by similar changes in NaPi-2b protein abundance and in rates of inorganic phosphate (Pi) uptake. OBJECTIVE: We tested the hypothesis that luminal fructose regulates NaPi-2b. DESIGN: We perfused into the intestine fructose, glucose, and nonmetabolizable or poorly transported glucose analogs as well as phlorizin. RESULTS: NaPi-2b mRNA concentrations and Pi uptake rates in fructose perfused intestines were approximately 30% of those in glucose and its analogs. NaPi-2b inhibition by fructose is specific because the mRNA abundance and activity of the fructose transporter GLUT5 (glucose transporter 5) increased with fructose perfusion, whereas those of other transporters were independent of the perfusate. Plasma Pi after 4 h of perfusion was independent of the perfusate, probably because normal kidneys can maintain normophosphatemia. Inhibiting glucose-6-phosphatase, another fructose-responsive gene, with tungstate or vanadate nonspecifically inhibited NaPi-2b mRNA expression and Pi uptake in both glucose- or fructose-perfused intestines. The AMP kinase (AMPK)-activator AICAR (5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside) enhanced and the fatty acid synthase-AMPK inhibitor C75 (3-carboxy-4-octyl-2-methylenebutyrolactone trans-4-carboxy-5-octyl-3-methylenebutyrolactone) prevented fructose inhibition of NaPi-2b but had no effect on expression of other transporters. NaPi-2b expression decreased markedly with age and was inhibited by fructose in all age groups. CONCLUSIONS: Energy levels in enterocytes may play a role in NaPi-2b inhibition by luminal fructose. Consumption of fructose that supplies approximately 10% of caloric intake by Americans clearly affects absorption of Pi and may promote Pi homeostasis in patients with impaired renal function. PMID- 18400730 TI - Influence of moderate energy restriction and seafood consumption on bone turnover in overweight young adults. AB - BACKGROUND: Overweight and obesity are increasing in young adults. However, moderate energy restriction aimed at lowering body weight may promote bone turnover and bone loss. Inclusion of fish or fish oils in a weight-loss diet may attenuate these adverse skeletal effects. OBJECTIVE: We examined the effects of incorporating fish or fish oil into an energy-restricted diet on bone turnover markers in young overweight adults. DESIGN: While following a strict hypoenergetic (-30% relative to estimated requirements) diet for 8 wk, 276 overweight men and women [body mass index (in kg/m(2)): 27.5-32.5; age: 20-40 y) were randomly assigned to 1 of 4 dietary groups: sunflower-oil capsules (3 g/d; control), cod (3 x 150 g/wk), salmon (3 x 150 g/wk), and fish-oil capsules (3 g/d). Body weight, bone biomarkers, and 25-hydroxyvitamin D were measured at baseline and endpoint. Data were analyzed with repeated-measures analysis of variance and general linear models. RESULTS: The mean (+/-SD) weight loss was 5.14 +/- 3.0 kg (5.8% +/- 3.2% body weight) during the 8 wk in the 4 dietary groups combined. Urinary N-telopeptides of type I collagen and serum C-terminal telopeptide of type I collagen increased (P < 0.05), whereas serum osteocalcin (but not bone-specific alkaline phosphatase) decreased (P < 0.05) from baseline to endpoint. Increased fish or fish-oil consumption had no effect (P > 0.1) on the changes in bone markers induced by weight loss. In contrast, increased salmon consumption increased serum 25-hydroxyvitamin D (P < 0.01). CONCLUSIONS: A nutritionally adequate but energy-restricted diet, with different contents of n-3 fatty acids, which resulted in modest weight loss, unfavorably altered bone turnover markers in young overweight adults. Such changes were not prevented by increased fish or fish-oil consumption. PMID- 18400731 TI - Comparative effects of dietary supplementation with red grape juice and vitamin E on production of superoxide by circulating neutrophil NADPH oxidase in hemodialysis patients. AB - BACKGROUND: Atherosclerotic cardiovascular disease is the most common cause of death among hemodialysis patients; it has been attributed to increased oxidative stress, dyslipidemia, malnutrition, and chronic inflammation. Activation of neutrophils is a well-recognized feature in dialysis patients, and superoxide anion production by neutrophil NADPH oxidase may contribute significantly to oxidative stress. OBJECTIVE: The aim of the study was to compare the effects of dietary supplementation with concentrated red grape juice (RGJ), a source of polyphenols, and vitamin E on neutrophil NADPH oxidase activity and other cardiovascular risk factors in hemodialysis patients. DESIGN: Thirty-two patients undergoing hemodialysis were recruited and randomly assigned to groups to receive dietary supplementation with RGJ, vitamin E, or both or a control condition without supplementation or placebo. Blood was obtained at baseline and on days 7 and 14 of treatment. RESULTS: RGJ consumption but not vitamin E consumption reduced plasma concentrations of total cholesterol and apolipoprotein B and increased those of HDL cholesterol. Both RGJ and vitamin E reduced plasma concentrations of oxidized LDL and ex vivo neutrophil NADPH oxidase activity. These effects were intensified when the supplements were used in combination; in that case, reductions in the inflammatory biomarkers intercellular adhesion molecule 1 and monocyte chemoattractant protein 1 also were observed. CONCLUSIONS: Regular ingestion of concentrated RGJ by hemodialysis patients reduces neutrophil NADPH-oxidase activity and plasma concentrations of oxidized LDL and inflammatory biomarkers to a greater extent than does that of vitamin E. This effect of RGJ consumption may favor a reduction in cardiovascular risk. PMID- 18400732 TI - No unique role for fructose sweeteners in obesity or cardiorenal disease. PMID- 18400733 TI - Time to get specific about dietary carbohydrates, affected populations, and diseases. PMID- 18400734 TI - The interaction between intestinal fatty acid-binding protein 2 polymorphism and delta-6 desaturase activity in obese children. PMID- 18400735 TI - An error in the US Department of Agriculture nutrient database results in vitamin A values that are 6 times too high. PMID- 18400736 TI - Homocysteine, vitamins, and vascular disease prevention: more negative results. PMID- 18400737 TI - Assessment of vitamin D in population-based studies. Preface. PMID- 18400738 TI - Vitamin D deficiency: a worldwide problem with health consequences. AB - Vitamin D deficiency is now recognized as a pandemic. The major cause of vitamin D deficiency is the lack of appreciation that sun exposure in moderation is the major source of vitamin D for most humans. Very few foods naturally contain vitamin D, and foods that are fortified with vitamin D are often inadequate to satisfy either a child's or an adult's vitamin D requirement. Vitamin D deficiency causes rickets in children and will precipitate and exacerbate osteopenia, osteoporosis, and fractures in adults. Vitamin D deficiency has been associated with increased risk of common cancers, autoimmune diseases, hypertension, and infectious diseases. A circulating level of 25-hydroxyvitamin D of >75 nmol/L, or 30 ng/mL, is required to maximize vitamin D's beneficial effects for health. In the absence of adequate sun exposure, at least 800-1000 IU vitamin D3/d may be needed to achieve this in children and adults. Vitamin D2 may be equally effective for maintaining circulating concentrations of 25 hydroxyvitamin D when given in physiologic concentrations. PMID- 18400739 TI - Blood biomarkers of vitamin D status. AB - In the past quarter century, more than 50 metabolites of vitamin D have been described. To date, only a few of these have been quantified in blood, but this has widened our understanding of the pathologic role that altered vitamin D metabolism plays in the development of diseases of calcium homeostasis. Currently, awareness is growing of the prevalence of vitamin D insufficiency in the general population in association with an increased risk of several diseases. However, for many researchers, it is not clear which vitamin D metabolites should be quantified and what the information gained from such an analysis tells us. Only 2 metabolites, namely, 25-hydroxyvitamin D [25(OH)D] and 1,25 dihydroxyvitamin D [1,25(OH)2D], have received the greatest attention. Of these, the need for measuring serum 1,25(OH)2D is limited, and this metabolite should therefore not be considered as part of the standard vitamin D testing regimen. On the other hand, serum 25(OH)D provides the single best assessment of vitamin D status and thus should be the only vitamin D assay typically performed. Currently, numerous formats exist for measuring serum 25(OH)D concentrations, each with its own advantages and disadvantages. This article reviews the currently available methods for serum 25(OH)D quantitation and considers important issues such as whether both the D2 and the D3 forms of the vitamin should be assayed, whether total or free concentrations are most important, and what measures should be taken to ensure the fidelity of the measurements. PMID- 18400740 TI - Vitamin D in foods: development of the US Department of Agriculture database. AB - Specific data on vitamin D2 and vitamin D3 are needed to enable the assessment of vitamin D dietary intake. These forms of the vitamin can occur in foods, both naturally or from fortification. The Nutrient Data Laboratory at the Beltsville Human Nutrition Research Center, Agricultural Research Service, US Department of Agriculture collaborated with vitamin D experts in an analytic project with 2 major goals: 1) to review and develop methods for analyzing a variety of food items for vitamin D content and 2) to sample and analyze foods considered to be major contributors of vitamin D. During 2007, analysts from up to 6 laboratories compared methods, made modifications in some cases, and validated results with quality-control samples of similar food types in preparation for the analysis of sampled foods. The Nutrient Data Laboratory has prioritized foods for analysis and has identified the following as important contributors of vitamin D: finfish and shellfish, naturally occurring sources, and fortified foods such as milk, calcium-fortified orange juice, breakfast cereals, American cheese, margarines, and yogurt. A nationwide multistage sampling plan was designed and conducted to select and procure representative sample units of all such foods. After analysis of these food samples and review of the results, acceptable values for vitamin D2 and D3 will be disseminated in the National Nutrient Database for Standard Reference (Internet: http://www.ars.usda.gov/nutrientdata). PMID- 18400741 TI - Sunlight exposure assessment: can we accurately assess vitamin D exposure from sunlight questionnaires? AB - The purpose of this review is to summarize the peer-reviewed literature in relation to sunlight exposure assessment and the validity of using sunlight exposure questionnaires to quantify vitamin D status. There is greater variability in personal ultraviolet (UV) light exposure as the result of personal behavior than as the result of ambient UV light exposure. Although statistically significant, the correlation coefficients for the relation between personal report of sun exposure and ambient UV light measured by dosimetry (assessment of radiation dose) are relatively low. Moreover, the few studies to assess the relation between sunlight measures and serum 25-hydroxyvitamin D show low correlations. These low correlations may not be surprising given that personal factors like melanin content in skin and age also influence cutaneous synthesis of vitamin D. In summary, sunlight exposure questionnaires currently provide imprecise estimates of vitamin D status. Research should be directed to develop more objective, nonintrusive, and economical measures of sunlight exposure to quantify personal vitamin D status. PMID- 18400742 TI - Vitamin D assessment in population-based studies: a review of the issues. AB - In the past decade, research on the relation between vitamin D exposure and disease in population-based studies has increased exponentially. These studies have involved measurement of vitamin D exposure by means of several methods: blood assays, self-reported dietary and supplemental intakes, and sunlight exposure questionnaires or diaries. As with all exposure measurements, researchers must consider the validity of their assessment tools for capturing vitamin D exposure. The purpose of this article is to summarize our current understanding of the various approaches to measuring vitamin D status within populations as reviewed at the 2007 Experimental Biology symposium, "Assessment of Vitamin D in Population-Based Studies." In summary, serum 25-hydroxyvitamin D is the accepted biomarker for short-term vitamin D status, but estimates of long term dietary and supplemental intakes of vitamin D and long-term sunlight exposure may be the most logistically feasible indicators of lifetime vitamin D exposure in population-based studies. Also discussed are issues investigators should consider when analyzing relations between vitamin D exposure and disease outcomes in population-based studies as well as research avenues that need further exploration. The best method for assessing vitamin D status in population based studies will depend primarily on the research question asked and the critical window of vitamin D exposure hypothesized to be most important. PMID- 18400743 TI - Heme oxygenase-1-derived carbon monoxide induces the Mycobacterium tuberculosis dormancy regulon. AB - The mechanisms that allow Mycobacterium tuberculosis (Mtb) to persist in human tissue for decades and to then abruptly cause disease are not clearly understood. Regulatory elements thought to assist Mtb to enter such a state include the heme two-component sensor kinases DosS and DosT and the cognate response regulator DosR. We have demonstrated previously that O(2), nitric oxide (NO), and carbon monoxide (CO) are regulatory ligands of DosS and DosT. Here, we show that in addition to O(2) and NO, CO induces the complete Mtb dormancy (Dos) regulon. Notably, we demonstrate that CO is primarily sensed through DosS to induce the Dos regulon, whereas DosT plays a less prominent role. We also show that Mtb infection of macrophage cells significantly increases the expression, protein levels, and enzymatic activity of heme oxygenase-1 (HO-1, the enzyme that produces CO), in an NO-independent manner. Furthermore, exploiting HO-1(+/+) and HO-1(-/-) bone marrow-derived macrophages, we demonstrate that physiologically relevant levels of CO induce the Dos regulon. Finally, we demonstrate that increased HO-1 mRNA and protein levels are produced in the lungs of Mtb-infected mice. Our data suggest that during infection, O(2), NO, and CO are being sensed concurrently rather than independently via DosS and DosT. We conclude that CO, a previously unrecognized host factor, is a physiologically relevant Mtb signal capable of inducing the Dos regulon, which introduces a new paradigm for understanding the molecular basis of Mtb persistence. PMID- 18400744 TI - Spatial and temporal dynamics of jasmonate synthesis and accumulation in Arabidopsis in response to wounding. AB - A new metabolite profiling approach combined with an ultrarapid sample preparation procedure was used to study the temporal and spatial dynamics of the wound-induced accumulation of jasmonic acid (JA) and its oxygenated derivatives in Arabidopsis thaliana. In addition to well known jasmonates, including hydroxyjasmonates (HOJAs), jasmonoyl-isoleucine (JA-Ile), and its 12-hydroxy derivative (12-HOJA-Ile), a new wound-induced dicarboxyjasmonate, 12 carboxyjasmonoyl-l-isoleucine (12-HOOCJA-Ile) was discovered. HOJAs and 12-HOOCJA Ile were enriched in the midveins of wounded leaves, strongly differentiating them from the other jasmonate metabolites studied. The polarity of these oxylipins at physiological pH correlated with their appearance in midveins. When the time points of accumulation of different jasmonates were determined, JA levels were found to increase within 2-5 min of wounding. Remarkably, these changes occurred throughout the plant and were not restricted to wounded leaves. The speed of the stimulus leading to JA accumulation in leaves distal to a wound is at least 3 cm/min. The data give new insights into the spatial and temporal accumulation of jasmonates and have implications in the understanding of long distance wound signaling in plants. PMID- 18400745 TI - Hyaluronan production in synoviocytes as a consequence of viral infections: HAS1 activation by Epstein-Barr virus and synthetic double- and single-stranded viral RNA analogs. AB - One of the hallmarks of arthritis is swollen joints containing unusually high quantities of hyaluronan. Intact hyaluronan molecules facilitate cell migration by acting as ligands for CD44. Hyaluronan degradation products, readily formed at sites of inflammation, also fuel inflammatory processes. Irrespective of whether viruses could be a cause of rheumatoid arthritis, there is clear evidence that links viral infections to this debilitating disease. For this study, live Epstein Barr virus and a number of double- and single-stranded synthetic viral analogs were tested for their effectiveness as activators of hyaluronan (HA) synthesis. As shown herein, Epstein-Barr virus-treated fibroblast-like synoviocytes significantly increase HA production and release. Real time reverse transcription PCR data show that HAS1 mRNA levels are significantly elevated in virus-treated cells, whereas mRNA levels for the genes HAS2 and HAS3 remain unchanged. As to the mechanism of virus-induced HAS1 transcription, data are presented that imply that among the double- and single-stranded polynucleotides tested, homopolymeric polycytidylic structures are the most potent inducers of HAS1 transcription and HA release, whereas homopolymeric polyinosinic acid is without effect. Analyses of virus-induced signal cascades, utilizing chemical inhibitors of MAPK and overexpressing mutated IKK and IkappaB, revealed that the MAPK p38 as well as the transcription factor NF-kappaB are essential for virus-induced activation of HAS1. The presented data implicate HAS1 as the culprit in unfettered HA release and point out targets in virus-induced signaling pathways that might allow for specific interventions in cases of unwanted and uncontrolled HA synthesis. PMID- 18400746 TI - Avian IgY binds to a monocyte receptor with IgG-like kinetics despite an IgE-like structure. AB - An ancestor of avian IgY was the evolutionary precursor of mammalian IgG and IgE, and present day chicken IgY performs the function of human IgG despite having the domain structure of human IgE. The kinetics of IgY binding to its receptor on a chicken monocyte cell line, MQ-NCSU, were measured, the first time that the binding of a non-mammalian antibody to a non-mammalian cell has been investigated (k(+1) = 1.14 +/- 0.46 x 10(5) mol(-1)sec(-1), k(-1) = 2.30 +/- 0.14 x 10(-3) s( 1), and K(a) = 4.95 x 10(7) m(-1)). This is a lower affinity than that recorded for mammalian IgE-high affinity receptor interactions (Ka approximately 10(10) m( 1)) but is within the range of mammalian IgG-high affinity receptor interactions (human: Ka approximately 10(8)-10(9) m(-1) mouse: Ka approximately 10(7)-10(8) m( 1). IgE has an extra pair of immunoglobulin domains when compared with IgG. Their presence reduces the dissociation rate of IgE from its receptor 20-fold, thus contributing to the high affinity of IgE. To assess the effect of the equivalent domains on the kinetics of IgY binding, IgY-Fc fragments with and without this domain were cloned and expressed in mammalian cells. In contrast to IgE, their presence in IgY has little effect on the association rate and no effect on dissociation. Whatever the function of this extra domain pair in avian IgY, it has persisted for at least 310 million years and has been co-opted in mammalian IgE to generate a uniquely slow dissociation rate and high affinity. PMID- 18400747 TI - STAT6 Inhibits TGF-beta1-mediated Foxp3 induction through direct binding to the Foxp3 promoter, which is reverted by retinoic acid receptor. AB - It has been shown that transforming growth factor beta1 (TGF-beta1) is critical in the generation of CD4(+)CD25(+)Foxp3(+)-inducible regulatory T cells (iTregs) from naive CD4(+)T cells. However, in contrast to natural Tregs, TGF-beta1 induced iTregs rapidly lose both Foxp3 expression and suppression activity. We found that TGF-beta1-induced Foxp3 levels were maintained by the addition of the anti-interleukin 4 (IL-4) antibody or by STAT6 gene deletion. Thus, IL-4 is an important suppressor of Foxp3 induction, and T helper 2 development is a major cause for the disappearance of iTreg during long culture. Using promoter analysis in EL4 cells and primary T cells, we identified a silencer region containing a STAT6 binding site. STAT6 binding to this site reduced TGF-beta1-mediated Foxp3 promoter activation and chromatin modification. Retinoic acid has also been shown to suppress loss of Foxp3 induced by TGF-beta1. Retinoic acid in the presence of TGF-beta1 reduced STAT6 binding to the Foxp3 promoter and enhanced histone acetylation, thereby reverting the effect of IL-4. We propose that antagonistic agents for neutralizing IL-4 could be a novel strategy to facilitate inducible Treg cell generation and the promotion of tolerance in Th2-dominated diseases such as allergy. PMID- 18400749 TI - Three novel collagen VI chains, alpha4(VI), alpha5(VI), and alpha6(VI). AB - We report the identification of three new collagen VI genes at a single locus on human chromosome 3q22.1. The three new genes are COL6A4, COL6A5, and COL6A6 that encode the alpha4(VI), alpha5(VI), and alpha6(VI) chains. In humans, the COL6A4 gene has been disrupted by a chromosome break. Each of the three new collagen chains contains a 336-amino acid triple helix flanked by seven N-terminal von Willebrand factor A-like domains and two (alpha4 and alpha6 chains) or three (alpha5 chain) C-terminal von Willebrand factor A-like domains. In humans, mRNA expression of COL6A5 is restricted to a few tissues, including lung, testis, and colon. In contrast, the COL6A6 gene is expressed in a wide range of fetal and adult tissues, including lung, kidney, liver, spleen, thymus, heart, and skeletal muscle. Antibodies to the alpha6(VI) chain stained the extracellular matrix of human skeletal and cardiac muscle, lung, and the territorial matrix of articular cartilage. In cell transfection and immunoprecipitation experiments, mouse alpha4(VI)N6-C2 chain co-assembled with endogenous alpha1(VI) and alpha2(VI) chains to form trimeric collagen VI molecules that were secreted from the cell. In contrast, alpha5(VI)N5-C1 and alpha6(VI)N6-C2 chains did not assemble with alpha1(VI) and alpha2(VI) chains and accumulated intracellularly. We conclude that the alpha4(VI)N6-C2 chain contains all the elements necessary for trimerization with alpha1(VI) and alpha2(VI). In summary, the discovery of three additional collagen VI chains doubles the collagen VI family and adds a layer of complexity to collagen VI assembly and function in the extracellular matrix. PMID- 18400748 TI - The kinetics of p53 activation versus cyclin E accumulation underlies the relationship between the spindle-assembly checkpoint and the postmitotic checkpoint. AB - Although cells can exit mitotic block aberrantly by mitotic slippage, they are prevented from becoming tetraploids by a p53-dependent postmitotic checkpoint. Intriguingly, disruption of the spindle-assembly checkpoint also compromises the postmitotic checkpoint. The precise mechanism of the interplay between these two pivotal checkpoints is not known. We found that after prolonged nocodazole exposure, the postmitotic checkpoint was facilitated by p53. We demonstrated that although disruption of the mitotic block by a MAD2-binding protein promoted slippage, it did not influence the activation of p53. Both p53 and its downstream target p21(CIP1/WAF1) were activated at the same rate irrespective of whether the spindle-assembly checkpoint was enforced or not. The accelerated S phase entry, as reflected by the premature accumulation of cyclin E relative to the activation of p21(CIP1/WAF1), is the reason for the uncoupling of the postmitotic checkpoint. In support of this hypothesis, forced premature mitotic exit with a specific CDK1 inhibitor triggered DNA replication without affecting the kinetics of p53 activation. Finally, replication after checkpoint bypass was boosted by elevating the level of cyclin E. These observations indicate that disruption of the spindle-assembly checkpoint does not directly influence p53 activation, but the shortening of the mitotic arrest allows cyclin E-CDK2 to be activated before the accumulation of p21(CIP1/WAF1). These data underscore the critical relationship between the spindle-assembly checkpoint and the postmitotic checkpoint in safeguarding chromosomal stability. PMID- 18400750 TI - 2-o-phosphorylation of xylose and 6-o-sulfation of galactose in the protein linkage region of glycosaminoglycans influence the glucuronyltransferase-I activity involved in the linkage region synthesis. AB - Sulfated glycosaminoglycans (GAGs), including heparan sulfate and chondroitin sulfate, are synthesized on the so-called common GAG-protein linkage region (GlcUAbeta1-3Galbeta1-3Galbeta1-4Xylbeta1-O-Ser) of core proteins, which is formed by the stepwise addition of monosaccharide residues by the respective specific glycosyltransferases. Glucuronyltransferase-I (GlcAT-I) is the key enzyme that completes the synthesis of this linkage region, which is a prerequisite for the conversion of core proteins to functional proteoglycans bearing GAGs. The Xyl and Gal residues in the linkage region can be modified by phosphorylation and sulfation, respectively, although the biological significance of these modifications remains to be clarified. Here we present evidence that these modifications can significantly influence the catalytic activity of GlcAT I. Enzyme assays showed that the synthetic substrates, Gal-Gal-Xyl(2-O-phosphate) O-Ser and Gal-Gal(6-O-sulfate)-Xyl(2-O-phosphate)-O-Ser, served as better substrates than the unmodified compound, whereas Gal(6-O-sulfate)-Gal-Xyl(2-O phosphate)-O-Ser exhibited no acceptor activity. The crystal structure of the catalytic domain of GlcAT-I with UDP and Gal-Gal(6-O-sulfate)-Xyl(2-O-phosphate) O-Ser bound revealed that the Xyl(2-O-phosphate)-O-Ser is disordered and the 6-O sulfate forms interactions with Gln(318) from the second GlcAT-I monomer in the dimeric enzyme. The results indicate the possible involvement of these modifications in the processing and maturation of the growing linkage region oligosaccharide required for the assembly of GAG chains. PMID- 18400751 TI - Conserved conformational changes in the ATPase cycle of human Hsp90. AB - The dimeric molecular chaperone Hsp90 is required for the activation and stabilization of hundreds of substrate proteins, many of which participate in signal transduction pathways. The activation process depends on the hydrolysis of ATP by Hsp90. Hsp90 consists of a C-terminal dimerization domain, a middle domain, which may interact with substrate protein, and an N-terminal ATP-binding domain. A complex cycle of conformational changes has been proposed for the ATPase cycle of yeast Hsp90, where a critical step during the reaction requires the transient N-terminal dimerization of the two protomers. The ATPase cycle of human Hsp90 is less well understood, and significant differences have been proposed regarding key mechanistic aspects. ATP hydrolysis by human Hsp90alpha and Hsp90beta is 10-fold slower than that of yeast Hsp90. Despite these differences, our experiments suggest that the underlying enzymatic mechanisms are highly similar. In both cases, a concerted conformational rearrangement involving the N-terminal domains of both subunits is controlling the rate of ATP turnover, and N-terminal cross-talk determines the rate-limiting steps. Furthermore, similar to yeast Hsp90, the slow ATP hydrolysis by human Hsp90s can be stimulated up to over 100-fold by the addition of the co-chaperone Aha1 from either human or yeast origin. Together, our results show that the basic principles of the Hsp90 ATPase reaction are conserved between yeast and humans, including the dimerization of the N-terminal domains and its regulation by the repositioning of the ATP lid from its original position to a catalytically competent one. PMID- 18400752 TI - Histone acetylation and methylation at sites initiating divergent polycistronic transcription in Trypanosoma cruzi. AB - Trypanosomes are ancient eukaryotic parasites in which the protein-coding genes, organized in large polycistronic clusters on both strands, are transcribed from as yet unidentified promoters. In an effort to reveal transcriptional initiation sites, we examined the Trypanosoma cruzi genome for histone modification patterns shown to be linked to active genes in various organisms. Here, we show that acetylated and methylated histones were found to be enriched at strand switch regions of divergent gene arrays, not at convergent clusters or intra- and intergenic regions within clusters. The modified region showed a bimodular profile with two peaks centered over the 5'-regions of the gene pair flanking the strand switch region. This pattern, which demarcates polycistronic transcription units originating from bidirectional initiation sites, is likely to be common in kinetoplastid parasites as well as in other organisms with polycistronic transcription. In contrast, no acetylation was found at promoters of the highly expressed rRNA and spliced leader genes or satellite DNA or at tested retrotransposonal elements. These results reveal, for the first time, the presence of specific epigenetic marks in T. cruzi with potential implications for transcriptional regulation; they indicate that both histone modifications and bidirectional transcription are evolutionarily conserved. PMID- 18400753 TI - Altered dimer interface decreases stability in an amyloidogenic protein. AB - Amyloidoses are devastating and currently incurable diseases in which the process of amyloid formation causes fatal cellular and organ damage. The molecular mechanisms underlying amyloidoses are not well known. In this study, we address the structural basis of immunoglobulin light chain amyloidosis, which results from deposition of light chains produced by clonal plasma cells. We compare light chain amyloidosis protein AL-09 to its wild-type counterpart, the kappaI O18/O8 light chain germline. Crystallographic studies indicate that both proteins form dimers. However, AL-09 has an altered dimer interface that is rotated 90 degrees from the kappaI O18/O8 dimer interface. The three non-conservative mutations in AL-09 are located within the dimer interface, consistent with their role in the decreased stability of this amyloidogenic protein. Moreover, AL-09 forms amyloid fibrils more quickly than kappaI O18/O8 in vitro. These results support the notion that the increased stability of the monomer and delayed fibril formation, together with a properly formed dimer, may be protective against amyloidogenesis. This could open a new direction into rational drug design for amyloidogenic proteins. PMID- 18400754 TI - High mobility group protein HMGB2 is a critical regulator of plasmodium oocyst development. AB - The sexual cycle of Plasmodium is required for transmission of malaria from mosquitoes to mammals, but how parasites induce the expression of genes required for the sexual stages is not known. We disrupted the Plasmodium yoelii gene encoding high mobility group nuclear factor hmgb2, which encodes a DNA-binding protein potentially implicated in transcriptional regulation of malaria gene expression. We investigated its function in vivo in the vertebrate and invertebrate hosts. Deltapyhmgb2 parasites develop into gametocytes but have drastic impairment of oocyst formation. A global transcriptome analysis of the Deltapyhmgb2 parasites identified approximately 30 genes whose expression is down regulated in the Deltapyhmgb2 parasites. These genes are conserved in all malaria species, and more than 90% of these genes show a peak of mRNA expression at the gametocyte stage. Surprisingly, the transcripts coding for the Plasmodium berghei orthologues of those genes are stored and translated in the ookinete stage. Therefore, sexual stage protein expression appears to be both transcriptionally and translationally regulated with Plasmodium HMGB2 acting as an important regulator of malaria sexual stage gene expression. PMID- 18400755 TI - X-ray structure of the [FeFe]-hydrogenase maturase HydE from Thermotoga maritima. AB - Maturation of the [FeFe]-hydrogenase active site depends on at least the expression of three gene products called HydE, HydF, and HydG. We have solved the high resolution structure of recombinant, reconstituted S-adenosine-L-methionine dependent HydE from Thermotoga maritima. Besides the conserved [Fe(4)S(4)] cluster involved in the radical-based reaction, this HydE was reported to have a second [Fe(4)S(4)] cluster coordinated by three Cys residues. However, in our crystals, depending on the reconstitution and soaking conditions, this second cluster is either a [Fe(2)S(2)] center, with water occupying the fourth ligand site or is absent. We have carried out site-directed mutagenesis studies on the related HydE from Clostridium acetobutylicum, along with in silico docking and crystal soaking experiments, to define the active site region and three anion binding sites inside a large, positive cavity, one of which binds SCN(-) with high affinity. Although the overall triose-phosphate isomerase-barrel structure of HydE is very similar to that of biotin synthase, the residues that line the internal cavity are significantly different in the two enzymes. PMID- 18400756 TI - Molecular chaperones and the assembly of the prion Ure2p in vitro. AB - The protein Ure2 from Saccharomyces cerevisiae possesses prion properties at the origin of the [URE3] trait. In vivo, a high molecular weight form of inactive Ure2p is associated to [URE3]. The faithful and continued propagation of [URE3]is dependent on the expression levels of molecular chaperones from the Hsp100, -70, and -40 families; however, so far, their role is not fully documented. Here we investigate the effects of molecular chaperones from the Hsp40, Hsp70, Hsp90, and Hsp100 families and the chaperonin CCT/Tric on the assembly of full-length Ure2p. We show that Hsp104p greatly stimulates Ure2p aggregation, whereas Ssa1p, Ydj1p, Sis1p, and Hsp82p inhibit aggregation to different extents. The nature of the high molecular weight Ure2p species that forms in the presence of the different molecular chaperones and their nucleotide dependence is described. We show that Hsp104p favors the aggregation of Ure2p into non-fibrillar high molecular weight particles, whereas Ssa1p, Ydj1p, Sis1p, and Hsp82p sequester Ure2p in spherical oligomers. Using fluorescently labeled full-length Ure2p and Ure2p-(94-354) and fluorescence polarization, we show that Ssa1p binding to Ure2p is ATP-dependent, whereas that of Hsp104p is not. We also show that Ssa1p preferentially interacts with the N-terminal domain of Ure2p that is critical for prion propagation, whereas Ydj1p preferentially interacts with the C-terminal domain of the protein, and we discuss the significance of this observation. Finally, the affinities of Ssa1p, Ydj1p, and Hsp104p for Ure2p are determined. Our in vitro observations bring new insight into the mechanism by which molecular chaperones influence the propagation of [URE3]. PMID- 18400757 TI - The same primary structure of the prion protein yields two distinct self propagating states. AB - The question of whether distinct self-propagating structures could be formed within the same amino acid sequence in the absence of external cofactors or templates has important implications for a number of issues, including the origin of prion strains and the engineering of smart, self-assembling peptide-based biomaterials. In the current study, we showed that chemically identical prion protein can give rise to conformationally distinct, self-propagating amyloid structures in the absence of cellular cofactors, post-translational modification, or PrP(Sc)-specified templates. Even more surprising, two self-replicating states were produced under identical solvent conditions, but under different shaking modes. Individual prion conformations were inherited by daughter fibrils in seeding experiments conducted under alternative shaking modes, illustrating the high fidelity of fibrillation reactions. Our study showed that the ability to acquire conformationally different self-propagating structures is an intrinsic ability of protein fibrillation and strongly supports the hypothesis that conformational variation in self-propagating protein states underlies prion strain diversity. PMID- 18400758 TI - The Hsp60-(p.V98I) mutation associated with hereditary spastic paraplegia SPG13 compromises chaperonin function both in vitro and in vivo. AB - We have previously reported the association of a mutation (c.292G > A/p.V98I) in the human HSPD1 gene that encodes the mitochondrial Hsp60 chaperonin with a dominantly inherited form of hereditary spastic paraplegia. Here, we show that the purified Hsp60-(p.V98I) chaperonin displays decreased ATPase activity and exhibits a strongly reduced capacity to promote folding of denatured malate dehydrogenase in vitro. To test its in vivo functions, we engineered a bacterial model system that lacks the endogenous chaperonin genes and harbors two plasmids carrying differentially inducible operons with human Hsp10 and wild-type Hsp60 or Hsp10 and Hsp60-(p.V98I), respectively. Ten hours after shutdown of the wild-type chaperonin operon and induction of the Hsp60-(p.V98I)/Hsp10 mutant operon, bacterial cell growth was strongly inhibited. No globally increased protein aggregation was observed, and microarray analyses showed that a number of genes involved in metabolic pathways, some of which are essential for robust aerobic growth, were strongly up-regulated in Hsp60-(p.V98I)-expressing bacteria, suggesting that the growth arrest was caused by defective folding of some essential proteins. Co-expression of Hsp60-(p.V98I) and wild-type Hsp60 exerted a dominant negative effect only when the chaperonin genes were expressed at relatively low levels. Based on our in vivo and in vitro data, we propose that the major effect of heterozygosity for the Hsp60-(p.V98I) mutation is a moderately decreased activity of chaperonin complexes composed of mixed wild-type and Hsp60-(p.V98I) mutant subunits. PMID- 18400759 TI - Sex-dependent effect of BAG1 in ameliorating motor deficits of Huntington disease transgenic mice. AB - The pathogenesis of Huntington disease (HD) is attributed to the misfolding of huntingtin (htt) caused by an expanded polyglutamine (polyQ) domain. Considerable effort has been devoted to identifying molecules that can prevent or reduce htt misfolding and the associated neuropathology. Although overexpression of chaperones is known to reduce htt cytotoxicity in cellular models, only modest protection is seen with Hsp70 overexpression in HD mouse models. Because the activity of Hsp70 is modulated by co-chaperones, an interesting issue is whether the in vivo effects of chaperones on polyQ protein toxicity are dependent on other modulators. In the present study, we focused on BAG1, a co-chaperone that interacts with Hsp70 and regulates its activity. Of htt mice expressing the N171 82Q mutant, we found that male N171-82Q mice show a greater deficit in rotarod performance than female N171-82Q mice. This sex-dependent motor deficit was improved by crossing N171-82Q mice with transgenic mice overexpressing BAG1 in neurons. Transgenic BAG1 also reduces the levels of mutant htt in synaptosomal fraction of male HD mice. Overexpression of BAG1 augmented the effects of Hsp70 by reducing aggregation of mutant htt in cultured cells and improving neurite outgrowth in htt-transfected PC12 cells. These findings suggest that the effects of chaperones on HD pathology are influenced by both their modulators and sex dependent factors. PMID- 18400760 TI - Novel chimeras of botulinum neurotoxins A and E unveil contributions from the binding, translocation, and protease domains to their functional characteristics. AB - Hyperexcitability disorders of cholinergically innervated muscles are treatable with botulinum neurotoxin (BoNT) A. The seven serotypes (A-G) potently block neurotransmission by binding to presynaptic receptors, undergoing endocytosis, transferring to the cytosol, and inactivating proteins essential for vesicle fusion. Although BoNT/A and BoNT/E cleave SNAP-25, albeit at distinct sites, BoNT/E blocks neurotransmission faster and more potently. To identify the domains responsible for these characteristics, the C-terminal heavy chain portions of BoNT/A and BoNT/E were exchanged to create chimeras AE and EA. After high yield expression in Escherichia coli, these single chain chimeras were purified by two step chromatography and activated by conversion to disulfide-linked dichains. In vitro, each entered neurons, cleaved SNAP-25, and blocked neuromuscular transmission while causing flaccid paralysis in vivo. Acidification-dependent translocation of the light chain to the cytosol occurred more rapidly for BoNT/E and EA than for BoNT/A and AE because the latter pair remained susceptible for longer to inhibitors of the vesicular proton pump, and BoNT/A proved less sensitive. The receptor-binding and protease domains do not seem to be responsible for the speeds of intoxication; rather the N-terminal halves of their heavy chains are implicated, with dissimilar rates of cytosolic transfer of the light chains being due to differences in pH sensitivity. AE produced the most persistent muscle weakening and therefore has therapeutic potential. Thus, proof of principle is provided for tailoring the pharmacological properties of these toxins by protein engineering. PMID- 18400761 TI - Interactions between the yeast SM22 homologue Scp1 and actin demonstrate the importance of actin bundling in endocytosis. AB - The yeast SM22 homologue Scp1 has previously been shown to act as an actin bundling protein in vitro. In cells, Scp1 localizes to the cortical actin patches that form as part of the invagination process during endocytosis, and its function overlaps with that of the well characterized yeast fimbrin homologue Sac6p. In this work we have used live cell imaging to demonstrate the importance of key residues in the Scp1 actin interface. We have defined two actin binding domains within Scp1 that allow the protein to both bind and bundle actin without the need for dimerization. Green fluorescent protein-tagged mutants of Scp1 also indicate that actin localization does not require the putative phosphorylation site Ser-185 to be functional. Deletion of SCP1 has few discernable effects on cell growth and morphology. However, we reveal that scp1 deletion is compensated for by up-regulation of Sac6. Furthermore, Scp1 levels are increased in the absence of sac6. The presence of compensatory pathways to up-regulate Sac6 or Scp1 levels in the absence of the other suggest that maintenance of sufficient bundling activity is critical within the cell. Analysis of cortical patch assembly and movement during endocytosis reveals a previously undetected role for Scp1 in movement of patches away from the plasma membrane. Additionally, we observe a dramatic increase in patch lifetime in a strain lacking both sac6 and scp1, demonstrating the central role played by actin-bundling proteins in the endocytic process. PMID- 18400763 TI - Crystal structures of the 70-kDa heat shock proteins in domain disjoining conformation. AB - The 70-kDa heat shock proteins (Hsp70s) are highly conserved ATP-dependent molecular chaperones composed of an N-terminal nucleotide binding domain (NBD) and a C-terminal protein substrate binding domain (SBD) in a bilobate structure. Interdomain communication and nucleotide-dependent structural motions are critical for Hsp70 chaperone functions. Our understanding of these functions remains elusive due to insufficient structural information on intact Hsp70s that represent the different states of the chaperone cycle. We report here the crystal structures of DnaK from Geobacillus kaustophilus HTA426 bound with ADP-Mg(2+) P(i) at 2.37A and the 70-kDa heat shock cognate protein from Rattus norvegicus bound with ADP-P(i) at 3.5A(.) The NBD and SBD in these structures are significantly separated from each other, and they might depict the ADP-bound conformation. Moreover, a Trp reporter was introduced at the potential interface region between NBD and the interdomain linker of GkDnaK to probe environmental changes. Results from fluorescence measurements support the notion that substrate binding enhances the domain-disjoining behavior of Hsp70 chaperones. PMID- 18400762 TI - ASAP3 is a focal adhesion-associated Arf GAP that functions in cell migration and invasion. AB - ASAP3, an Arf GTPase-activating protein previously called DDEFL1 and ACAP4, has been implicated in the pathogenesis of hepatocellular carcinoma. We have examined in vitro and in vivo functions of ASAP3 and compared it to the related Arf GAP ASAP1 that has also been implicated in oncogenesis. ASAP3 was biochemically similar to ASAP1: the pleckstrin homology domain affected function of the catalytic domain by more than 100-fold; catalysis was stimulated by phosphatidylinositol 4,5-bisphosphate; and Arf1, Arf5, and Arf6 were used as substrates in vitro. Like ASAP1, ASAP3 associated with focal adhesions and circular dorsal ruffles. Different than ASAP1, ASAP3 did not localize to invadopodia or podosomes. Cells, derived from a mammary carcinoma and from a glioblastoma, with reduced ASAP3 expression had fewer actin stress fiber, reduced levels of phosphomyosin, and migrated more slowly than control cells. Reducing ASAP3 expression also slowed invasion of mammary carcinoma cells. In contrast, reduction of ASAP1 expression had no effect on migration or invasion. We propose that ASAP3 functions nonredundantly with ASAP1 to control cell movement and may have a role in cancer cell invasion. In comparing ASAP1 and ASAP3, we also found that invadopodia are dispensable for the invasive behavior of cells derived from a mammary carcinoma. PMID- 18400765 TI - Analysis of potential predictors of depression among coronary heart disease risk factors including heart rate variability, markers of inflammation, and endothelial function. AB - AIMS: We investigated the relationship between autonomic nervous system balance, systemic immune activation, endothelial dysfunction, and depression in patients free of coronary heart disease (CHD) with increased CHD risk. METHODS AND RESULTS: Depression status (Beck Depression Inventory, BDI), selected CHD risk factors, inflammation markers, measures of heart rate variability (HRV), and indices of endothelial function (flow-mediated dilation, FMD) were evaluated in 415 subjects free of CHD, diabetes mellitus, and other life-threatening conditions, with at least two CHD risk factors among the following: older age, male gender, current smoking, hypertension, and dislipidaemia. Overall, 51.7% of the participants were males, aged 57.6 +/- 8.8 years on average (minimum 30, maximum 70). Almost half were hypertensive, 43.9% were dyslipidemic, 30.4% current smokers, and 23.1% showed a depressive symptomatology (BDI > or = 10). Logistic regression showed that, as compared with non-depressed individuals and after adjustment for age, gender, and hypertension, depressive subjects were significantly more likely to be smokers, to have higher total cholesterol, higher C-reactive protein, and Interleukin-6. In addition, depressed subjects were more likely to have altered HRV and their FMD was severely impaired (adjusted odds ratio of 1% increase = 0.72; 95% CI: 0.61-0.86). CONCLUSION: Our data indicate an independent association between depression and impaired HRV, systemic inflammatory, and endothelial function. These mechanisms play a role not only in the complication of advanced forms of disease, but also promote and/or accelerate the early disease and connect depression and CHD. PMID- 18400764 TI - Global Risk Profile Verification in Patients with Venous Thromboembolism (GRIP VTE) in 5 Gulf countries. AB - BACKGROUND: The Global Risk Profile Verification in Patients with Venous Thromboembolism was the first prospective multicenter registry conducted in Arabian Gulf countries to explore the epidemiology of venous thromboembolic (VTE) disorders and to provide data on diagnosis and disease management. METHODS: Data on 242 patients with confirmed VTE were submitted between September 2003 and November 2003 from 28 contributing hospitals in the Arabian Gulf region. Differences between groups were assessed by the chi(2) test or Fisher exact test for categorical variables. The Student's t test was used for testing proportions. RESULTS: The frequency of VTE cases is deep vein thrombosis (DVT), 187 (77.27%); pulmonary embolism (PE), 35 (14.46%); and DVT with PE, 20 (8.26%). The most common symptoms of DVT and DVT/PE patients were calf pain (72%), calf swelling (63.8%), and localized tenderness (52.2%). The most common symptoms in patients with PE alone and DVT/PE were dyspnea (83.6%), thoracic pain (69.1%), and cough (40%). Risk factors for VTE were immobilization (41.3%), age >65 years (28.9%), a history of VTE (20.7%), and trauma (19%). Among surgical interventions, orthopedic procedures induced the greatest number of VTE cases, followed by general surgery and gynecological procedures. Low-molecular-weight heparins were chosen to treat 33.7% of DVT cases, whereas unfractionated heparin was used in 21.9% of cases. CONCLUSION: VTE remains a common problem in medical and surgical patients in the Arabian Gulf states. Recognition of the common risk factors is of extreme importance to implement the appropriate prophylactic strategy according to the published guidelines. PMID- 18400766 TI - Purification and characterization of a copper-containing amine oxidase from Mycobacterium sp. strain JC1 DSM 3803 grown on benzylamine. AB - A bacterial semicarbazide-sensitive amine oxidase (SSAO) was purified and characterized from Mycobacterium sp. strain JC1 DSM 3803 grown on benzylamine. During the purification procedures, the enzyme was tending to aggregate and exhibited heterogeneity in native PAGE. The heterogeneous forms having amine oxidase (AO) activity could be separated by their native molecular weights using gel-filtration chromatography. Most of the AOs behaved as dimers (M(r) 150,000) composed of a 75-kDa subunit, but some aggregated to form tetramers (M(r) 300,000). Besides their native molecular weight, subunit composition and V(max) value, both forms (dimer and tetramer) have almost identical biochemical properties (e.g. subunit size, optimum pH and temperature, activation energy, K(m) value on benzylamine, substrate and inhibitor specificities). When AO activity was observed by activity staining, the best-oxidized substrate was benzylamine, although the AO also oxidized tyramine and histamine. The AO was strongly inhibited by semicarbazide and isoniazid, but KCN did not affect its activity. The purified enzyme was shown to contain 2.39 mol of copper per mole of subunit, but there were no evidences of topaquinone co-factor involvement, when tested by absorption spectrum analysis and redox-cycling staining for quinoprotein detection. PMID- 18400767 TI - 222RN and 220RN concentrations measured in various natural honey samples by using nuclear track detectors and resulting radiation doses to the members of the rural populations in Morocco. AB - Radon (222Rn) and thoron (220Rn) concentrations were measured in sixteen natural honey material samples collected from different regions in Morocco by using CR-39 and LR-115 type II solid state nuclear track detectors (SSNTDs). The concentrations of these radionuclides were also measured in nectar solutions corresponding to the studied honey samples. The measured concentrations of 222Rn and 220Rn in honey samples ranged from (2.3 +/- 0.2) to (8.1 +/- 0.6) Bq.l(-1) and (1.8 +/- 0.1) to (3.9 +/- 0.3) Bq.l(-1), respectively. Committed equivalent doses due to annual intakes of 222Rn were evaluated in the human gastrointestinal tract compartments of adult members of the Moroccan populations from the ingestion of studied honey samples. The influence of the target tissue mass and activities due to 222Rn on the annual committed equivalent doses in the compartments of the human gastrointestinal tract was investigated. PMID- 18400768 TI - Exposure assessment of workers to airborne PCDD/Fs, PCBs and PAHs at an electric arc furnace steelmaking plant in the UK. AB - Occupational exposure studies were undertaken at a UK electric arc furnace (EAF) steelmaking plant to investigate the exposure of workers via inhalation to dioxins, polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene (B[a]P). Surveys were undertaken in areas including the melting shop, the casting department and a furnace control cabin. The highest concentrations of dioxins and PCBs were found inside the melting shop nearby EAFs, whereas dioxin and PCB concentrations in the casting department and inside the control cabin were significantly lower. Risk characterization was carried out by comparing the daily intake of dioxins and PCBs through inhalation with the recommended tolerable daily intake (TDI). Health risk assessments were also carried out by combining exposure data with inhalation cancer potency factors to quantify the cancer risk. For the most exposed category of workers (melting shop workers), the estimated daily intake via inhalation was 0.35 pg WHO TEQ kg(-1) body weight (bw) in the worst case scenario. Considering that the average UK adult exposure to dioxins from the diet is 1.8 pg WHO-TEQ kg(-1) bw day(-1), the results indicated that the estimated daily intake of dioxins via inhalation at the EAF would not result in the recommended range of the TDI (1-4 pg WHO-TEQ kg(-1) bw day(-1)) being exceeded. Cancer risks for a 40-year occupational exposure period were determined by multiplying the inhalation dose by the inhalation cancer potency factor for 2,3,7,8-tetrachlorodibenzo-p-dioxin. For melting shop workers, cancer risks from exposure to dioxins and PCBs ranged from 2.05 x 10(-5) to 7.54 x 10(-5). Under most regulatory programmes, excess cancer risks between 1.0 x 10(-4) and 1.0 x 10(-6) indicate an acceptable range of excess cancer risk, suggesting a limited risk from dioxin exposure for workers in the EAF plant. For the calculation of excess cancer risks, no account has been taken of the protection provided by protective respiratory equipment worn by EAF workers. If personal protective equipments were taken into consideration, it is likely that the excess cancer risks for EAF workers would have been lower and considered as negligible. The highest concentrations of PAHs were found in the melting shop and the casting areas of the plant. In the melting shop area, B[a]P concentrations ranged from 1.4 to 24.5 ng m(-3), with a mean value of 7 ng m(-3). No workplace exposure limits have been published by the Health and Safety Executive in the UK for PAHs; however, the B[a]P concentrations found were below the limit value of 150 ng m(-3) (8-h time-weighted average) specified for workplace exposure in France. Exposure assessment of workers to PAHs via inhalation was carried out by calculating a potential cancer risk considering a 40-year occupational exposure period and B[a]Peq concentrations. Estimated cancer risks for the most exposed category of workers (i.e. melting shop workers) ranged from 3.66 x 10(-6) to 1.64 x 10(-5). The cancer risks determined in this study were well within an acceptable range of excess cancer risk of 1.0 x 10(-4) to 1.0 x 10(-6), specified by the US Environmental Protection Agency. PMID- 18400769 TI - Diabetic control and risk of tuberculosis: a cohort study. AB - Diabetes mellitus is associated with tuberculosis. A cohort of 42,116 clients aged 65 years or more, enrolled at 18 Elderly Health Service centers in Hong Kong in 2000, were followed up prospectively through the territory-wide tuberculosis registry for development of tuberculosis from 3 months after enrollment to December 31, 2005, by use of their identity card numbers as unique identifier. The effects of diabetes mellitus and diabetic control on tuberculosis risk were assessed with adjustment for sociodemographic and other background variables. Diabetes mellitus was associated with a modest increase in the risk of active, culture-confirmed, and pulmonary (with or without extrapulmonary involvement) but not extrapulmonary (with or without pulmonary involvement) tuberculosis, with adjusted hazard ratios of 1.77 (95% confidence interval: 1.41, 2.24), 1.91 (95% confidence interval: 1.45, 2.52), 1.89 (95% confidence interval: 1.48, 2.42), and 1.00 (95% confidence interval: 0.54, 1.86), respectively. Diabetic subjects with hemoglobin A1c <7% at enrollment were not at increased risk. Among diabetic subjects, higher risks of active, culture-confirmed, and pulmonary but not extrapulmonary tuberculosis were observed with baseline hemoglobin A1c > or =7% (vs. <7%), with adjusted hazard ratios of 3.11 (95% confidence interval: 1.63, 5.92), 3.08 (95% confidence interval: 1.44, 6.57), 3.63 (95% confidence interval: 1.79, 7.33), and 0.77 (95% confidence interval: 0.18, 3.35), respectively. PMID- 18400771 TI - Fast network component analysis (FastNCA) for gene regulatory network reconstruction from microarray data. AB - MOTIVATION: Recently developed network component analysis (NCA) approach is promising for gene regulatory network reconstruction from microarray data. The existing NCA algorithm is an iterative method which has two potential limitations: computational instability and multiple local solutions. The subsequently developed NCA-r algorithm with Tikhonov regularization can help solve the first issue but cannot completely handle the second one. Here we develop a novel Fast Network Component Analysis (FastNCA) algorithm which has an analytical solution that is much faster and does not have the above limitations. RESULTS: Firstly FastNCA is compared to NCA and NCA-r using synthetic data. The reconstruction of FastNCA is more accurate than that of NCA-r and comparable to that of properly converged NCA. FastNCA is not sensitive to the correlation among the input signals, while its performance does degrade a little but not as dramatically as that of NCA. Like NCA, FastNCA is not very sensitive to small inaccuracies in a priori information on the network topology. FastNCA is about several tens times faster than NCA and several hundreds times faster than NCA-r. Then, the method is applied to real yeast cell-cycle microarray data. The activities of the estimated cell-cycle regulators by FastNCA and NCA-r are compared to the semi-quantitative results obtained independently by Lee et al. (2002). It is shown here that there is a greater agreement between the results of FastNCA and Lee's, which is represented by the ratio 23/33, than that between the results of NCA-r and Lee's, which is 14/33. AVAILABILITY: Software and supplementary materials are available from http://www.eee.hku.hk/~cqchang/FastNCA.htm PMID- 18400770 TI - Independent predictive accuracy of classical electrocardiographic criteria in the diagnosis of paroxysmal atrioventricular reciprocating tachycardias in patients without pre-excitation. AB - AIMS: In patients without pre-excitation, the differential diagnosis of paroxysmal atrioventricular (AV) reciprocating tachycardias consists mainly of atrioventricular nodal re-entrant tachycardias (AVNRTs) and AV reciprocating tachycardias (AVRTs) through a concealed bypass. Our purpose was to validate the diagnostic accuracy of a predictive logistic model using classical electrocardiographic (ECG) criteria. METHODS AND RESULTS: We included 470 patients who underwent an electrophysiological study for paroxysmal, regular, and narrow-QRS complex tachycardia without pre-excitation in sinus rhythm. The ECG recordings were reviewed for the presence of the following: (i) pseudo r' deflection (V1) and/or pseudo s-wave (inferior leads), (ii) identifiable P-wave after the QRS complex, (iii) QRS alternans, and (iv) repolarization abnormalities during tachycardia. We performed a cross-validation method using the first 300 patients to develop a logistic model to predict the tachycardia diagnosis. The model was validated through the remaining 170 patients. The invasive study demonstrated AVNRT in 314 patients and AVRT in 156 patients. The presence of pseudo r' deflection and/or pseudo s-wave, a visible P-wave after the QRS complex, and QRS alternans were selected by a stepwise multiple logistic regression analysis as predictors for the diagnosis of AVNRT. The application of the model in the validation group showed a shrinkage prediction factor of 3%. Diagnostic probabilities for both tachycardia mechanisms depending on every combination of selected ECG criteria were >75% in 70% of the patients. CONCLUSION: The presence of pseudo r' deflection and/or pseudo s-wave, an identifiable P-wave after the QRS, and QRS alternans during tachycardia permit us to derive a reliable logistic model to predict the mechanism of paroxysmal AVRT in patients without pre-excitation. Precise probabilities for a correct diagnosis associated with every combination of those classical ECG criteria are presented. PMID- 18400772 TI - EPO-KB: a searchable knowledge base of biomarker to protein links. AB - The knowledge base EPO-KB (Empirical Proteomic Ontology Knowledge Base) is based on an OWL ontology that represents current knowledge linking mass-to-charge (m/z) ratios to proteins on multiple platforms including Matrix Assisted Laser/Desorption Ionization (MALDI) and Surface Enhanced Laser/Desorption Ionization (SELDI)--Time of Flight (TOF). At present, it contains information on m/z ratio to protein links that were extracted from 120 published research papers. It has a web interface that allows researchers to query and retrieve putative proteins that correspond to a user-specified m/z ratio. EPO-KB also allows automated entry of additional m/z ratio to protein links and is expandable to the addition of gene to protein and protein to disease links. AVAILABILITY: http://www.dbmi.pitt.edu/EPO-KB PMID- 18400773 TI - MedEvi: retrieving textual evidence of relations between biomedical concepts from Medline. AB - Search engines running on MEDLINE abstracts have been widely used by biologists to find publications that are related to their research. The existing search engines such as PubMed, however, have limitations when applied for the task of seeking textual evidence of relations between given concepts. The limitations are mainly due to the problem that the search engines do not effectively deal with multi-term queries which may imply semantic relations between the terms. To address this problem, we present MedEvi, a novel search engine that imposes positional restriction on occurrences matching multi-term queries, based on the observation that terms with semantic relations which are explicitly stated in text are not found too far from each other. MedEvi further identifies additional keywords of biological and statistical significance from local context of matching occurrences in order to help users reformulate their queries for better results. AVAILABILITY: http://www.ebi.ac.uk/tc-test/textmining/medevi/ PMID- 18400774 TI - A system for generating transcription regulatory networks with combinatorial control of transcription. AB - We have developed a new software system, REgulatory Network generator with COmbinatorial control (RENCO), for automatic generation of differential equations describing pre-transcriptional combinatorics in artificial regulatory networks. RENCO has the following benefits: (a) it explicitly models protein-protein interactions among transcription factors, (b) it captures combinatorial control of transcription factors on target genes and (c) it produces output in Systems Biology Markup Language (SBML) format, which allows these equations to be directly imported into existing simulators. Explicit modeling of the protein interactions allows RENCO to incorporate greater mechanistic detail of the transcription machinery compared to existing models and can provide a better assessment of algorithms for regulatory network inference. AVAILABILITY: RENCO is a C++ command line program, available at http://sourceforge.net/projects/renco/ PMID- 18400775 TI - Nominal and effective dosimetry of silica nanoparticles in cytotoxicity assays. AB - Because of their small size and large specific surface area (SA), insoluble nanoparticles are almost not affected by the gravitational force and are generally formulated in stable suspensions or sols. This raises, however, a potential difficulty in in vitro assay systems in which cells adhering to the bottom of a culture vessel may not be exposed to the majority of nanoparticles in suspension. J. G. Teeguarden et al., 2007, Toxicol. Sci. 95, 300-312 have recently addressed this issue theoretically, emphasizing the need to characterize the effective dose (mass or number or SA dose of particles that affect the cells) which, according to their model based on sedimentation and gravitation forces, might only represent a very small fraction of the nominal dose. We hypothesized, in contrast, that because of convection forces that usually develop in sols, the majority of the particles may reach the target cells and exert their potential toxicity. To address this issue, we exposed three different cell lines (A549 epithelial cells, EAHY926 endothelial cells, and J774 monocyte-macrophages) to a monodisperse suspension of Stober silica nanoparticles (SNP) in three different laboratories. Four different end points (lacticodehydrogenase [LDH] release, LDH cell content, tetrazolium salt (MTT), and crystal violet staining) were used to assess the cell response to nanoparticles. We found, in all cell lines and for all end points, that the cellular response was determined by the total mass/number/SA of particles as well as their concentration. Practically, for a given volume of dispersion, both parameters are of course intimately interdependent. We conclude that the nominal dose remains the most appropriate metric for in vitro toxicity testing of insoluble SNP dispersed in aqueous medium. This observation has important bearings on the experimental design and the interpretation of in vitro toxicological studies with nanoparticles. PMID- 18400777 TI - Reticulated platelets as a screening test to identify thrombocytopenia aetiology. AB - BACKGROUND: Thrombocytopenia is a common haematological abnormality and no simple diagnostic test is available to diagnose thrombocytopenia pathogenesis. AIM: To evaluate sensitivity and specificity of reticulated platelets (RP) as a diagnostic test for thrombocytopenia with increased thrombopoietic activity. DESIGN: Prospective observational study in thrombocytopenic patients. METHODS: A direct, whole-blood, dual-labelling flow cytometric method was used. Direct, whole-blood double coverage was achieved using a monoclonal anti-glycoprotein (GP)-III antibody (CD61 PerCP) for platelet identification and thiazole orange (Retic-count) as platelet mARN stain. RESULTS: RP were measured in 101 thrombocytopenic patients and 104 non-thrombocytopenic controls. The mean RP percentage in 60 thrombocytopenic patients with no increased thrombopoietic activity was 7.5% (CI for 95%: 5.2-9.7) and RP absolute number was 3.2 x 10(9)/l (CI for 95%: 2.1-4.3). The mean RP percentage in 41 thrombocytopenic patients with increased thrombopoietic activity was 30.3% (CI for 95%: 25.1-35.5) and RP absolute number was 6.2 (CI for 95%: 4.8-7.7). The RP percentage cut-off for a diagnosis of thrombocytopenia with increased thrombopoietic activity was 11% [sensitivity 93%, specificity 85%, positive predictive value (PPV) 83%, negative predictive value (NPV) 95%]. CONCLUSION: RP measurement by flow cytometry, directly from whole-blood, is a useful screening test to differentiate between thrombocytopenia with high or low thrombopoietic activity. A RP percentage in excess of 11%, has a high sensitivity and good specificity for a diagnosis of thrombocytopenia with increased thrombopoietic activity. PMID- 18400776 TI - A randomised controlled trial of intramuscular vs. intravenous antivenom for latrodectism--the RAVE study. AB - BACKGROUND: Widow spider-bite causes latrodectism and is associated with significant morbidity worldwide. Antivenom is given by both the intravenous (IV) and intramuscular (IM) routes and it is unclear which is more effective. AIM: To compare the effectiveness of IV vs. IM redback spider antivenom. DESIGN: Randomized controlled trial. METHODS: Patients with latrodectism were given either IV or IM antivenom according to a randomized double-dummy, double-blind protocol. The first antivenom treatment was followed by another identical treatment after two hours if required. The primary outcome was a clinically significant reduction in pain two hours after the last treatment. A fully Bayesian analysis was used to estimate the probability of the desired treatment effect, predetermined as an absolute difference of 20%. RESULTS: We randomly allocated 126 patients to receive antivenom IV (64) and IM (62). After antivenom treatment pain improved in 40/64(62%) in the IV group vs. 33/62(53%) in the IM group (+9%; 95% Credible Interval [CrI]: -8% to +26%). The probability of a difference greater than zero (IV superior) was 85% but the probability of a difference >20% was only 10%. In 55 patients with systemic effects, these improved in 58% after IV antivenom vs. 65% after IM antivenom (-8%; 95% CrI: -32% to +17%). Twenty-four hours after antivenom pain had improved in 84% in the IV group vs. 71% in the IM group (+13%; 95% CrI: -2% to +27%). A meta-analysis including data from a previous trial found no difference in the primary outcome between IV and IM administration. DISCUSSION: The difference between IV and IM routes of administration of widow spider antivenom is, at best, small and does not justify routinely choosing one route over the other. Furthermore, antivenom may provide no benefit over placebo. PMID- 18400778 TI - Detecting cis-regulatory binding sites for cooperatively binding proteins. AB - Several methods are available to predict cis-regulatory modules in DNA based on position weight matrices. However, the performance of these methods generally depends on a number of additional parameters that cannot be derived from sequences and are difficult to estimate because they have no physical meaning. As the best way to detect cis-regulatory modules is the way in which the proteins recognize them, we developed a new scoring method that utilizes the underlying physical binding model. This method requires no additional parameter to account for multiple binding sites; and the only necessary parameters to model homotypic cooperative interactions are the distances between adjacent protein binding sites in basepairs, and the corresponding cooperative binding constants. The heterotypic cooperative binding model requires one more parameter per cooperatively binding protein, which is the concentration multiplied by the partition function of this protein. In a case study on the bacterial ferric uptake regulator, we show that our scoring method for homotypic cooperatively binding proteins significantly outperforms other PWM-based methods where biophysical cooperativity is not taken into account. PMID- 18400779 TI - Inhibitory effect of a short Z-DNA forming sequence on transcription elongation by T7 RNA polymerase. AB - DNA sequences capable of forming unusual secondary structures can be a source of genomic instability. In some cases that instability might be affected by transcription, as recently shown for the Z-DNA forming sequence (CG)(14), which causes genomic instability both in mammalian cells and in bacteria, and this effect increases with its transcription. We have investigated the effect of this (CG)(14) sequence on transcription with T7 RNA polymerase in vitro. We detected partial transcription blockage within the sequence; the blockage increased with negative supercoiling of the template DNA. This effect was not observed in a control self-complementary sequence of identical length and base composition as the (CG)(14) sequence, when the purine-pyrimidine alternation required for Z-DNA formation was disrupted. These findings suggest that the inhibitory effect on T7 transcription results from Z-DNA formation in the (CG)(14) sequence rather than from an effect of the sequence composition or from hairpin formation in either the DNA or the RNA product. PMID- 18400781 TI - MultiPriDe: automated batch development of quantitative real-time PCR primers. AB - Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is a commonly employed gene expression quantification technique. This requires the development of appropriately targeted oligonucleotide primers, which necessitates the identification of ideal amplicons, development of optimized oligonucleotide sequences under most favorable pre-determined reaction conditions, and management of the resultant target-oligonucleotide pair information for each gene to be studied. The Primer3 utility exists for development of oligonucleotide primers and fills that role effectively. However, the manual process of identifying target sites and individually generating primers is inefficient and prone to user introduced error, especially when a large number of genes are to be examined. We have developed MultiPriDe (Multiple Primer Design), a Perl utility that accepts batch lists of Gene database identifiers, collects available intron and exon position data critical to qRT-PCR primer development, and supplies these sites as identified targets for the Primer3 utility. This automated 'gene to primer' procedure is coupled with a set of optimized hybridization conditions used by the Primer3 utility to maximize successful primer design. MultiPriDe and assembled repeat libraries are available upon request. Please direct requests to aziesel@emory.edu. PMID- 18400780 TI - Structural probing of the HIV-1 polypurine tract RNA:DNA hybrid using classic nucleic acid ligands. AB - The interactions of archetypical nucleic acid ligands with the HIV-1 polypurine tract (PPT) RNA:DNA hybrid, as well as analogous DNA:DNA, RNA:RNA and swapped hybrid substrates, were used to probe structural features of the PPT that contribute to its specific recognition and processing by reverse transcriptase (RT). Results from intercalative and groove-binding ligands indicate that the wild-type PPT hybrid does not contain any strikingly unique groove geometries and/or stacking arrangements that might contribute to the specificity of its interaction with RT. In contrast, neomycin bound preferentially and selectively to the PPT near the 5'(rA)(4):(dT)(4) tract and the 3' PPT-U3 junction. Nuclear magnetic resonance data from a complex between HIV-1 RT and the PPT indicate RT contacts within the same regions highlighted on the PPT by neomycin. These observations, together with the fact that the sites are correctly spaced to allow interaction with residues in the ribonuclease H (RNase H) active site and thumb subdomain of the p66 RT subunit, suggest that despite the long cleft employed by RT to make contact with nucleic acids substrates, these sites provide discrete binding units working in concert to determine not only specific PPT recognition, but also its orientation on the hybrid structure. PMID- 18400783 TI - Overexpression of human mitochondrial valyl tRNA synthetase can partially restore levels of cognate mt-tRNAVal carrying the pathogenic C25U mutation. AB - Phenotypic diversity associated with pathogenic mutations of the human mitochondrial genome (mtDNA) has often been explained by unequal segregation of the mutated and wild-type genomes (heteroplasmy). However, this simple hypothesis cannot explain the tissue specificity of disorders caused by homoplasmic mtDNA mutations. We have previously associated a homoplasmic point mutation (1624C>T) in MTTV with a profound metabolic disorder that resulted in the neonatal deaths of numerous siblings. Affected tissues harboured a marked biochemical defect in components of the mitochondrial respiratory chain, presumably due to the extremely low (<1%) steady-state levels of mt-tRNA(Val). In primary myoblasts and transmitochondrial cybrids established from the proband (index case) and offspring, the marked respiratory deficiency was lost and steady-state levels of the mutated mt-tRNA(Val) were greater than in the biopsy material, but were still an order of magnitude lower than in control myoblasts. We present evidence that the generalized decrease in steady-state mt-tRNA(Val) observed in the homoplasmic 1624C>T-cell lines is caused by a rapid degradation of the deacylated form of the abnormal mt-tRNA(Val). By both establishing the identity of the human mitochondrial valyl-tRNA synthetase then inducing its overexpression in transmitochondrial cell lines, we have been able to partially restore steady state levels of the mutated mt-tRNA(Val), consistent with an increased stability of the charged mt-tRNA. These data indicate that variations in the levels of VARS2L between tissue types and patients could underlie the difference in clinical presentation between individuals homoplasmic for the 1624C>T mutation. PMID- 18400782 TI - The mRNA encoding the yeast ARE-binding protein Cth2 is generated by a novel 3' processing pathway. AB - Microarray analyses of mRNAs over-expressed in strains lacking the nuclear exosome component Rrp6 identified the transcript encoding the ARE-binding protein Cth2, which functions in cytoplasmic mRNA stability. Subsequent northern analyses revealed that exosome mutants accumulate a 3'-extended transcript at the expense of the mature CTH2 mRNA. The 3' ends of the CTH2 mRNA were mapped to a [GU(3 5)](5) repeat, unlike any previously characterized polyadenylation site. CTH2 mRNA accumulation was not inhibited by mutations in 3'-cleavage and polyadenylation factors, Rna14, Rna15 and Pap1, which block accumulation of other mRNAs. The 3'-extended CTH2 pre-mRNA strongly accumulated in strains with mutations in the TRAMP4 polyadenylation complex or the Nrd1/Nab3/Sen1 complex, and contains multiple Nrd1 and Nab3 binding sites. CTH2 carries a consensus ARE element and levels of the pre-mRNA and mRNA were elevated by mutation of the ARE or inactivation of the nuclear 5'-exonuclease Rat1. We propose that CTH2 mRNA is processed from a 3'-extended primary transcript by the exosome, TRAMP and Nrd1/Nab3/Sen1 complexes. This unusual pathway may allow time for nuclear, ARE mediated regulation of CTH2 levels involving Rat1. PMID- 18400784 TI - The major form of hepatitis C virus alternate reading frame protein is suppressed by core protein expression. AB - Hepatitis C virus (HCV) is a human RNA virus encoding 10 proteins in a single open reading frame. In the +1 frame, an 'alternate reading frame' (ARF) overlaps with the core protein-encoding sequence and encodes the ARF protein (ARFP). Here, we investigated the molecular regulatory mechanisms of ARFP expression in HCV target cells. Chimeric HCV-luciferase reporter constructs derived from the infectious HCV prototype isolate H77 were transfected into hepatocyte-derived cell lines. Translation initiation was most efficient at the internal AUG codon at position 86/88, resulting in the synthesis of a truncated ARFP named (86/88)ARFP. Interestingly, (86/88)ARFP synthesis was markedly enhanced in constructs containing an inactivated core protein reading frame. This enhancement was reversed by co-expression of core protein in trans, demonstrating suppression of ARFP synthesis by HCV core protein. In conclusion, our results indicate that translation of ARFP occurs mainly by alternative internal initiation at position 86/88 and is regulated by HCV core protein expression. The suppression of ARFP translation by HCV core protein suggests that ARFP expression is inversely linked to the level of viral replication. These findings define key mechanisms regulating ARFP expression and set the stage for further studies addressing the function of ARFP within the viral life cycle. PMID- 18400785 TI - Testing the predictive power of the transtheoretical model of behavior change applied to dietary fat intake. AB - This study evaluated how well predictions from the transtheoretical model (TTM) generalized from smoking to diet. Longitudinal data were used from a randomized control trial on reducing dietary fat consumption in adults (n =1207) recruited from primary care practices. Predictive power was evaluated by making a priori predictions of the magnitude of change expected in the TTM constructs of temptation, pros and cons, and 10 processes of change when an individual transitions between the stages of change. Generalizability was evaluated by testing predictions based on smoking data. Three sets of predictions were made for each stage: Precontemplation (PC), Contemplation (C) and Preparation (PR) based on stage transition categories of no progress, progress and regression determined by stage at baseline versus stage at the 12-month follow-up. Univariate analysis of variance between stage transition groups was used to calculate the effect size [omega squared (omega(2))]. For diet predictions based on diet data, there was a high degree of confirmation: 92%, 95% and 92% for PC, C and PR, respectively. For diet predictions based on smoking data, 77%, 79% and 85% were confirmed, respectively, suggesting a moderate degree of generalizability. This study revised effect size estimates for future theory testing on the TTM applied to dietary fat. PMID- 18400786 TI - Language ability and verbal and nonverbal executive functioning in deaf students communicating in spoken English. AB - The internal use of language during problem solving is considered to play a key role in executive functioning. This role provides a means for self-reflection and self-questioning during the formation of rules and plans and a capacity to control and monitor behavior during problem-solving activity. Given that increasingly sophisticated language is required for effective executive functioning as an individual matures, it is likely that students with poor language abilities will have difficulties performing complex problem-solving tasks. The aim of this study was to investigate the relationship between language ability and verbal and nonverbal executive functioning in a group of deaf students who communicate using spoken English, as measured by their performance on two standardized tests of executive function: the Delis-Kaplan Executive Function System (D-KEFS) 20 Questions Test and the D-KEFS Tower Test. Expressive language ability accounted for more than 40% of variability in performance on the D-KEFS 20 Questions Test. There was no significant relationship between language ability and performance on the D-KEFS Tower Test. There was no relationship between language ability and familiarity with the specific problem-solving strategies of both D-KEFS Tests. Implications of the findings are discussed. PMID- 18400787 TI - The relationship between the reading and signing skills of deaf children in bilingual education programs. AB - This paper reports on one experiment in which we investigated the relationship between reading and signing skills. We administered a vocabulary task and a story comprehension task in Sign Language of the Netherlands and in written Dutch to a group of 87 deaf children from bilingual education programs. We found a strong and positive correlation between the scores obtained in the sign vocabulary task and the reading vocabulary task when age, short-term memory scores, and nonverbal intelligence scores were controlled for. In addition, a correlation was observed between the scores in the story comprehension tasks in Sign Language of the Netherlands and written Dutch but only when vocabulary scores for words and signs were not taken into account. The results are briefly discussed with reference to a model we recently proposed to describe lexical development for deaf children in bilingual education programs (Hermans, D., Knoors, H., Ormel, E., & Verhoeven, L., 2008). In addition, the implications of the results of the present study for previous studies on the relationship between reading and signing skills are discussed. PMID- 18400788 TI - Unequal rates of Y chromosome gene divergence during speciation of the family Ursidae. AB - Evolution of the bear family Ursidae is well investigated in terms of morphological, paleontological, and genetic features. However, several phylogenetic ambiguities occur within the subfamily Ursinae (the family Ursidae excluding the giant panda and spectacled bear), which may correlate with behavioral traits of female philopatry and male-biased dispersal which form the basis of the observed matriarchal population structure in these species. In the process of bear evolution, we investigate the premise that such behavioral traits may be reflected in patterns of variation among genes with different modes of inheritance: matrilineal mitochondrial DNA (mtDNA), patrilineal Y chromosome, biparentally inherited autosomes, and the X chromosome. In the present study, we sequenced 3 Y-linked genes (3,453 bp) and 4 X-linked genes (4,960 bp) and reanalyzed previously published sequences from autosome genes (2,347 bp) in ursid species to investigate differences in evolutionary rates associated with patterns of inheritance. The results describe topological incongruence between sex-linked genes and autosome genes and between nuclear DNA and mtDNA. In more ancestral branches within the bear phylogeny, Y-linked genes evolved faster than autosome and X-linked genes, consistent with expectations based on male-driven evolution. However, this pattern changes among branches leading to each species within the lineage of Ursinae whereby the evolutionary rates of Y-linked genes have fewer than expected substitutions. This inconsistency between more recent nodes of the bear phylogeny with more ancestral nodes may reflect the influences of sex-biased dispersal as well as molecular evolutionary characteristics of the Y chromosome, and stochastic events in species natural history, and phylogeography unique to ursine bears. PMID- 18400789 TI - A phylogenetic analysis of indel dynamics in the cotton genus. AB - Genome size evolution is a dynamic process involving counterbalancing mechanisms whose actions vary across lineages and over time. Whereas the primary mechanism of expansion, transposable element (TE) amplification, has been widely documented, the evolutionary dynamics of genome contraction have been less thoroughly explored. To evaluate the relative impact and evolutionary stability of the mechanisms that affect genome size, we conducted a phylogenetic analysis of indel rates for 2 genomic regions in 4 Gossypium genomes: the 2 coresident genomes (A(T) and D(T)) of tetraploid cotton and its model diploid progenitors, Gossypium arboreum (A) and Gossypium raimondii (D). We determined the rates of sequence gain or loss along each branch, partitioned by mechanism, and how these changed during species divergence. In general, there has been a propensity toward growth of the diploid genomes and contraction in the polyploid. Most of the size difference between the diploid species occurred prior to polyploid divergence and was largely attributable to TE amplification in the A/A(T) genome. After separating from the true parents of the polyploid genomes, both diploid genomes experienced slower sequence gain than in the ancestor, due to fewer TE insertions in the A genome and a combination of increased deletions and decreased TE insertions in the D genome. Both genomes of the polyploid displayed increased rates of deletion and decreased rates of insertion, leading to a rate of near stasis in D(T) and overall contraction in A(T) resulting in polyploid genome contraction. As expected, TE insertions contributed significantly to the genome size differences; however, intrastrand homologous recombination, although rare, had the most significant impact on the rate of deletion. Small indel data for the diploids suggest the possibility of a bias as the smaller genomes add less or delete more sequence through small indels than do the larger genomes, whereas data for the polyploid suggest increased sequence turnover in general (both as small deletions and small insertions). Illegitimate recombination, although not demonstrated to be a dominant mechanism of genome size change, was biased in the polyploid toward deletions, which may provide a partial explanation of polyploid genomic downsizing. PMID- 18400790 TI - Organellar RNA editing and plant-specific extensions of pentatricopeptide repeat proteins in jungermanniid but not in marchantiid liverworts. AB - The pyrimidine exchange type of RNA editing in land plant (embryophyte) organelles has largely remained an enigma with respect to its biochemical mechanisms, the underlying specificities, and its raison d'etre. Apparently arising with the earliest embryophytes, RNA editing is conspicuously absent in one clade of liverworts, the complex thalloid Marchantiidae. Several lines of evidence suggest that the large gene family of organelle-targeted RNA-binding pentatricopeptide repeat (PPR) proteins plays a fundamental role in the sequence specific editing of organelle transcripts. We here describe the identification of PPR protein genes with plant-specific carboxyterminal (C-terminal) sequence signatures (E, E+, and DYW domains) in ferns, lycopodiophytes, mosses, hornworts, and jungermanniid liverworts, one subclass of the basal most clade of embryophytes, on DNA and cDNA level. In contrast, we were unable to identify these genes in a wide sampling of marchantiid liverworts (including the phylogenetic basal genus Blasia)--taxa for which no RNA editing is observed in the organelle transcripts. On the other hand, we found significant diversity of this type of PPR proteins also in Haplomitrium, a genus with an extremely high rate of RNA editing and a phylogenetic placement basal to all other liverworts. Although the presence of modularly extended PPR proteins correlates well with organelle RNA editing, the now apparent complete loss of an entire gene family from one clade of embryophytes, the marchantiid liverworts, remains puzzling. PMID- 18400791 TI - Activity in face-responsive brain regions is modulated by invisible, attended faces: evidence from masked priming. AB - It is often assumed that neural activity in face-responsive regions of primate cortex correlates with conscious perception of faces. However, whether such activity occurs without awareness is still debated. Using functional magnetic resonance imaging (fMRI) in conjunction with a novel masked face priming paradigm, we observed neural modulations that could not be attributed to perceptual awareness. More specifically, we found reduced activity in several classic face-processing regions, including the "fusiform face area," "occipital face area," and superior temporal sulcus, when a face was preceded by a briefly flashed image of the same face, relative to a different face, even when 2 images of the same face differed. Importantly, unlike most previous studies, which have minimized awareness by using conditions of inattention, the present results occurred when the stimuli (the primes) were attended. By contrast, when primes were perceived consciously, in a long-lag priming paradigm, we found repetition related activity increases in additional frontal and parietal regions. These data not only demonstrate that fMRI activity in face-responsive regions can be modulated independently of perceptual awareness, but also document where such subliminal face-processing occurs (i.e., restricted to face-responsive regions of occipital and temporal cortex) and to what extent (i.e., independent of the specific image). PMID- 18400792 TI - A small world of neuronal synchrony. AB - A small-world network has been suggested to be an efficient solution for achieving both modular and global processing-a property highly desirable for brain computations. Here, we investigated functional networks of cortical neurons using correlation analysis to identify functional connectivity. To reconstruct the interaction network, we applied the Ising model based on the principle of maximum entropy. This allowed us to assess the interactions by measuring pairwise correlations and to assess the strength of coupling from the degree of synchrony. Visual responses were recorded in visual cortex of anesthetized cats, simultaneously from up to 24 neurons. First, pairwise correlations captured most of the patterns in the population's activity and, therefore, provided a reliable basis for the reconstruction of the interaction networks. Second, and most importantly, the resulting networks had small-world properties; the average path lengths were as short as in simulated random networks, but the clustering coefficients were larger. Neurons differed considerably with respect to the number and strength of interactions, suggesting the existence of "hubs" in the network. Notably, there was no evidence for scale-free properties. These results suggest that cortical networks are optimized for the coexistence of local and global computations: feature detection and feature integration or binding. PMID- 18400793 TI - Medial temporal lobe involvement in an implicit memory task: evidence of collaborating implicit and explicit memory systems from FMRI and Alzheimer's disease. AB - We used a prototype extraction task to assess implicit learning of a meaningful novel visual category. Cortical activation was monitored in young adults with functional magnetic resonance imaging. We observed occipital deactivation at test consistent with perceptually based implicit learning, and lateral temporal cortex deactivation reflecting implicit acquisition of the category's semantic nature. Medial temporal lobe (MTL) activation during exposure and test suggested involvement of explicit memory as well. Behavioral performance of Alzheimer's disease (AD) patients and healthy seniors was also assessed, and AD performance was correlated with gray matter volume using voxel-based morphometry. AD patients showed learning, consistent with preserved implicit memory, and confirming that AD patients' implicit memory is not limited to abstract patterns. However, patients were somewhat impaired relative to healthy seniors. Occipital and lateral temporal cortical volume correlated with successful AD patient performance, and thus overlapped with young adults' areas of deactivation. Patients' severe MTL atrophy precluded involvement of this region. AD patients thus appear to engage a cortically based implicit memory mechanism, whereas their relative deficit on this task may reflect their MTL disease. These findings suggest that implicit and explicit memory systems collaborate in neurologically intact individuals performing an ostensibly implicit memory task. PMID- 18400794 TI - Functional connectivity of human striatum: a resting state FMRI study. AB - Classically regarded as motor structures, the basal ganglia subserve a wide range of functions, including motor, cognitive, motivational, and emotional processes. Consistent with this broad-reaching involvement in brain function, basal ganglia dysfunction has been implicated in numerous neurological and psychiatric disorders. Despite recent advances in human neuroimaging, models of basal ganglia circuitry continue to rely primarily upon inference from animal studies. Here, we provide a comprehensive functional connectivity analysis of basal ganglia circuitry in humans through a functional magnetic resonance imaging examination during rest. Voxelwise regression analyses substantiated the hypothesized motor, cognitive, and affective divisions among striatal subregions, and provided in vivo evidence of a functional organization consistent with parallel and integrative loop models described in animals. Our findings also revealed subtler distinctions within striatal subregions not previously appreciated by task-based imaging approaches. For instance, the inferior ventral striatum is functionally connected with medial portions of orbitofrontal cortex, whereas a more superior ventral striatal seed is associated with medial and lateral portions. The ability to map multiple distinct striatal circuits in a single study in humans, as opposed to relying on meta-analyses of multiple studies, is a principal strength of resting state functional magnetic resonance imaging. This approach holds promise for studying basal ganglia dysfunction in clinical disorders. PMID- 18400796 TI - Molecular genetic basis of ribotyping. AB - Nearly 2,000 ribotyping-based studies exist, ranging from epidemiology to phylogeny and taxonomy. None precisely reveals the molecular genetic basis, with many incorrectly attributing detected polymorphisms to rRNA gene sequences. Based on in silico genomics, we demonstrate that ribotype polymorphisms result from sequence variability in neutral housekeeping genes flanking rRNA operons, with rRNA gene sequences serving solely as conserved, flank-linked tags. We also reveal that from such an informatics perspective, it is readily feasible a priori to design an interpretable ribotyping scheme for a genomically sequenced microbial species, and we discuss limitations to the basic restriction fragment length polymorphism-based method as well as alternate PCR ribotyping-based schemes. PMID- 18400795 TI - Central nervous system tuberculosis: pathogenesis and clinical aspects. AB - Tuberculosis of the central nervous system (CNS) is a highly devastating form of tuberculosis, which, even in the setting of appropriate antitubercular therapy, leads to unacceptable levels of morbidity and mortality. Despite the development of promising molecular diagnostic techniques, diagnosis of CNS tuberculosis relies largely on microbiological methods that are insensitive, and as such, CNS tuberculosis remains a formidable diagnostic challenge. Insights into the basic neuropathogenesis of Mycobacterium tuberculosis and the development of an appropriate animal model are desperately needed. The optimal regimen and length of treatment are largely unknown, and with the rising incidence of multidrug resistant strains of M. tuberculosis, the development of well-tolerated and effective antibiotics remains a continued need. While the most widely used vaccine in the world largely targets this manifestation of tuberculosis, the BCG vaccine has not fulfilled the promise of eliminating CNS tuberculosis. We put forth this review to highlight the current understanding of the neuropathogenesis of M. tuberculosis, to discuss certain epidemiological, clinical, diagnostic, and therapeutic aspects of CNS tuberculosis, and also to underscore the many unmet needs in this important field. PMID- 18400798 TI - Human bocavirus: passenger or pathogen in acute respiratory tract infections? AB - Human bocavirus (HBoV) is a newly identified virus tentatively assigned to the family Parvoviridae, subfamily Parvovirinae, genus Bocavirus. HBoV was first described in 2005 and has since been detected in respiratory tract secretions worldwide. Herein we review the literature on HBoV and discuss the biology and potential clinical impact of this virus. Most studies have been PCR based and performed on patients with acute respiratory symptoms, from whom HBoV was detected in 2 to 19% of the samples. HBoV-positive samples have been derived mainly from infants and young children. HBoV DNA has also been detected in the blood of patients with respiratory tract infection and in fecal samples of patients with diarrhea with or without concomitant respiratory symptoms. A characteristic feature of HBoV studies is the high frequency of coinciding detections, or codetections, with other viruses. Available data nevertheless indicate a statistical association between HBoV and acute respiratory tract disease. We present a model incorporating these somewhat contradictory findings and suggest that primary HBoV infection causes respiratory tract symptoms which can be followed by prolonged low-level virus shedding in the respiratory tract. Detection of the virus in this phase will be facilitated by other infections, either simply via increased sample cell count or via reactivation of HBoV, leading to an increased detection frequency of HBoV during other virus infections. We conclude that the majority of available HBoV studies are limited by the sole use of PCR diagnostics on respiratory tract secretions, addressing virus prevalence but not disease association. The ability to detect primary infection through the development of improved diagnostic methods will be of great importance for future studies seeking to assign a role for HBoV in causing respiratory illnesses. PMID- 18400797 TI - Respiratory viruses other than influenza virus: impact and therapeutic advances. AB - Though several antivirals have been developed and marketed to treat influenza virus infections, the development of antiviral agents with clinical activity against other respiratory viruses has been more problematic. Here we review the epidemiology of respiratory viral infections in immunocompetent and immunocompromised hosts, examine the evidence surrounding the currently available antivirals for respiratory viral infections other than influenza, highlight those that are in the pipeline, and discuss the hurdles for development of such agents. PMID- 18400802 TI - New aspects of neotropical polycystic (Echinococcus vogeli) and unicystic (Echinococcus oligarthrus) echinococcosis. AB - Of the four species of the genus Echinococcus (Cestoda) distinguished by biological and morphological characteristics, two species, E. vogeli and E. oligarthrus, occur widely in the Neotropics. Approximately 200 cases of polycystic echinococcosis (PE) have been recorded from 12 countries in South America. Following early proliferation of E. vogeli in the human host, typically in the liver, the metacestode usually spreads in the peritoneal and pleural cavities, and numerous organs may be invaded. The clinical characteristics of PE in 81 patients with sufficient information are reviewed. Type I disease consists of polycysts in the liver and abdominal cavity (37% of the patients had this characteristic); type II is similar to type I but also includes hepatic insufficiency (26%); type III consists of cysts in liver and chest (14%); type IV consists of cysts only in the mesenteries (16%); and type V consists of cysts calcified in liver and lung (4%). The percentage of patients with polycysts in the liver was 81%, and the percentage of patients with polycysts in the chest was 14%. PE is most ready diagnosed by geographic origin of the patient and by means of ultrasound or computerized tomography scanning showing cysts and calcifications. The highest mortality was for patients with type II disease, due to hepatic failure and its complications. There were five patients who died due to surgical accidents, whereas 35 cases had uncomplicated surgery. Twenty-three patients died of PE, making the total mortality 29% (23 of 78 cases). None of the 13 patients treated only with albendazole, the most efficacious treatment, was completely cured. PE represents a severe medical problem in South America. A reevaluation of the characteristics of the metacestode of E. oligarthrus indicated that it is unicystic. Only three human cases are known (two with infection in the orbit and one with infection in the heart). The metacestode of E. oligarthrus, in contrast with that of E. vogeli, consists of a spherical, fluid-filled vesicle that enlarges concentrically and is not known to undergo exogenous proliferation. PMID- 18400803 TI - Maternal use of fluconazole and risk of congenital malformations: a Danish population-based cohort study. AB - OBJECTIVES: Fluconazole is widely used for the treatment of candidiasis. Although the drug is also prescribed to pregnant women, data on the safety of use of fluconazole during pregnancy are limited. We examined the association between the maternal use of fluconazole during pregnancy and the risk of congenital malformations. PATIENTS AND METHODS: In this population-based cohort study in Northern Denmark, we included 1079 women who had a live birth or a stillbirth after the 20th week of gestation and who redeemed at least one prescription for fluconazole during the first trimester. The reference cohort comprised 170 453 pregnant women who redeemed no fluconazole prescription during pregnancy. The women were identified through the Danish Medical Birth Registry. Data on drug use, birth outcome and covariates were extracted from population-based healthcare databases. We used logistic regression to estimate the prevalence odds ratio (POR) for congenital malformations after fluconazole exposure, while adjusting for maternal smoking, parity, maternal age and concurrent prescriptions for antiepileptics or antidiabetics. RESULTS: Among 1079 women who filled a fluconazole prescription during the first trimester, 797 (74%) received a total of 150 mg of fluconazole, 235 (22%) received 300 mg of fluconazole, 24 (2%) received 350 mg of fluconazole and 23 (2%) received 600 mg of fluconazole. These women gave birth to 44 (4.1%) children with congenital malformations. The 170 453 women without fluconazole prescriptions gave birth to 6152 (3.6%) children with congenital malformations. For congenital malformations overall, the adjusted POR associated with the first-trimester fluconazole use was 1.0 (95% confidence interval: 0.8-1.4). CONCLUSIONS: We found no overall increased risk of congenital malformations after exposure to short-course treatment with fluconazole in early pregnancy. PMID- 18400804 TI - In vitro pharmacodynamics of novel rifamycin ABI-0043 against Staphylococcus aureus. AB - OBJECTIVES: ABI-0043 is a novel benzoxazinorifamycin derivative, which derives its potent bactericidal activity by the specific inhibition of bacterial RNA polymerase. We evaluated the in vitro pharmacodynamics and bactericidal activity of ABI-0043 against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA). METHODS: Using time kill studies at a wide range of concentrations of ABI-0043, we evaluated the killing activity against four clinical isolates of S. aureus over 24 h. An integrated pharmacokinetic/pharmacodynamic area measure was applied to all cfu data and was fitted to a Hill-type mathematical model to evaluate pharmacodynamics. RESULTS: Bacterial killing for ABI-0043 occurred rapidly and in a concentration-dependent manner. Bactericidal activity was achieved within 4 h at > or =16 x MIC against all isolates. Bacterial reductions were greatest at > or =64 x MIC against MRSA and MSSA isolates, as a >4 log(10) cfu/mL reduction was observed as early as 2 h, and sustained throughout 24 h. The pharmacodynamics of ABI-0043 was well described by a Hill-type model, with a steep sigmoidicity constant and a low EC(50) against all isolates. CONCLUSIONS: ABI-0043 displayed rapid and sustained bactericidal activity against S. aureus clinical isolates. ABI-0043 represents a promising antistaphylococcal agent to combat serious S. aureus infections. Further, pharmacokinetic, pharmacodynamic and in vivo studies are warranted to determine its ultimate place in antibacterial therapy. PMID- 18400805 TI - Antibiotic prescribing patterns in village health clinics across 10 provinces of Western China. AB - OBJECTIVES: To explore antibiotic prescribing patterns in rural areas across 10 provinces of Western China and to compare the patterns among these provinces. METHODS: About 20 125 prescriptions were collected from 680 primary health clinics in villages from 40 counties in 10 provinces of Western China. Percentage of prescriptions with antibiotics and number of antibiotics per 100 prescriptions were used as measurements of antibiotic utilization. RESULTS: The percentage of prescriptions with antibiotics was 48.43 (range: 41.12-57.47) in the study areas. There were 49 kinds of antibiotics prescribed in total, and 17 of them accounted for 90% of all usage. The number of antibiotics per 100 prescriptions was 54.62 (range: 43.78-69.56). CONCLUSIONS: The frequency and proportion of prescribed antibiotics in the rural areas of Western China are higher compared with the developed countries, and the patterns of antibiotic prescription differ greatly among provinces. The findings have important policy implications for recommendations on the utilization of antibiotics in China. PMID- 18400800 TI - The relationship between leishmaniasis and AIDS: the second 10 years. AB - To date, most Leishmania and human immunodeficiency virus (HIV) coinfection cases reported to WHO come from Southern Europe. Up to the year 2001, nearly 2,000 cases of coinfection were identified, of which 90% were from Spain, Italy, France, and Portugal. However, these figures are misleading because they do not account for the large proportion of cases in many African and Asian countries that are missed due to a lack of diagnostic facilities and poor reporting systems. Most cases of coinfection in the Americas are reported in Brazil, where the incidence of leishmaniasis has spread in recent years due to overlap with major areas of HIV transmission. In some areas of Africa, the number of coinfection cases has increased dramatically due to social phenomena such as mass migration and wars. In northwest Ethiopia, up to 30% of all visceral leishmaniasis patients are also infected with HIV. In Asia, coinfections are increasingly being reported in India, which also has the highest global burden of leishmaniasis and a high rate of resistance to antimonial drugs. Based on the previous experience of 20 years of coinfection in Europe, this review focuses on the management of Leishmania-HIV-coinfected patients in low-income countries where leishmaniasis is endemic. PMID- 18400801 TI - Anisakis simplex: from obscure infectious worm to inducer of immune hypersensitivity. AB - Infection of humans with the nematode worm parasite Anisakis simplex was first described in the 1960s in association with the consumption of raw or undercooked fish. During the 1990s it was realized that even the ingestion of dead worms in food fish can cause severe hypersensitivity reactions, that these may be more prevalent than infection itself, and that this outcome could be associated with food preparations previously considered safe. Not only may allergic symptoms arise from infection by the parasites ("gastroallergic anisakiasis"), but true anaphylactic reactions can also occur following exposure to allergens from dead worms by food-borne, airborne, or skin contact routes. This review discusses A. simplex pathogenesis in humans, covering immune hypersensitivity reactions both in the context of a living infection and in terms of exposure to its allergens by other routes. Over the last 20 years, several studies have concentrated on A. simplex antigen characterization and innate as well as adaptive immune response to this parasite. Molecular characterization of Anisakis allergens and isolation of their encoding cDNAs is now an active field of research that should provide improved diagnostic tools in addition to tools with which to enhance our understanding of pathogenesis and controversial aspects of A. simplex allergy. We also discuss the potential relevance of parasite products such as allergens, proteinases, and proteinase inhibitors and the activation of basophils, eosinophils, and mast cells in the induction of A. simplex-related immune hypersensitivity states induced by exposure to the parasite, dead or alive. PMID- 18400806 TI - Antibiotic use in 26 departments of internal medicine in 6 general hospitals in Israel: variability and contributing factors. AB - OBJECTIVES: Increased antibiotic consumption is associated with increased bacterial resistance worldwide. We aimed to analyse antibiotic consumption and potential contributory factors in internal medicine departments in Israel. METHODS: Data (2003-04) from 26 departments in 6 hospitals were retrieved. Defined daily doses (DDD)/100 bed-days were calculated for total antibiotic use and by antibiotic class. Patterns identified were correlated with 15 patients' and departmental variables by univariate and multivariate analyses. RESULTS: Total antibiotic consumption differed by a factor of 2.3 (115 DDD/100 bed-days to 49.1 DDD/100 bed-days) between the highest and lowest consuming departments. Antibiotic classes differed by a factor of 22.8 for macrolides, a factor of 20 for piperacillin/tazobactam, a factor of 17 for carbapenems, a factor of 13.3 for quinolones, a factor of 9 for vancomycin, a factor of 6.8 for amoxicillin/clavulanate, a factor of 6.6 for aminoglycosides, a factor of 5.3 for penicillins and a factor of 2.8 for cephalosporins. Even among departments within hospitals, there was a difference of up to 1.5-fold for total use and antibiotic class differences ranged between 2.5- and 7.2-fold for third- and fourth generation cephalosporins, despite similar Charlson scores and other patient variables. In the multivariate analysis, hospital affiliation and rate of 1 day hospitalization were the only significant variables predicting total antibiotic use, contributing 43% and 7.3%, respectively, to the variance. By antibiotic class, controlling for hospital affiliation, patients with neutropenia, lower respiratory tract infections and assisted ventilation were the most common significant contributors, ranging from 3.5% for quinolones to 7.7% for piperacillin/tazobactam. CONCLUSIONS: Patterns of antibiotic use vary widely among internal medicine departments in Israel, which cannot be explained by objective parameters related either to patients or wards. Ongoing monitoring and guideline formulation are needed to regulate antibiotic prescription. PMID- 18400807 TI - Maraviroc: integration of a new antiretroviral drug class into clinical practice. AB - Maraviroc (Pfizer's UK-427857, Selzentry or Celsentri outside the US) is the first agent in the new class of oral HIV-1 entry inhibitors to acquire FDA and EMEA approval. It is expected that this drug will be effective only in a subpopulation of HIV-1-infected people, namely those harbouring only the R5 virus. The wide use of this drug is currently hampered by the lack of a readily available R5 virus only determination test (tropism test) and by insufficient scientific insight into the dynamics of R5 and X4 viruses during infection. We discuss the challenges associated with the currently available assay, as well as the potential role of alternative assays. PMID- 18400799 TI - Cavitary pulmonary disease. AB - A pulmonary cavity is a gas-filled area of the lung in the center of a nodule or area of consolidation and may be clinically observed by use of plain chest radiography or computed tomography. Cavities are present in a wide variety of infectious and noninfectious processes. This review discusses the differential diagnosis of pathological processes associated with lung cavities, focusing on infections associated with lung cavities. The goal is to provide the clinician and clinical microbiologist with an overview of the diseases most commonly associated with lung cavities, with attention to the epidemiology and clinical characteristics of the host. PMID- 18400808 TI - Effect of nitrous oxide on plasma homocysteine and folate in patients undergoing major surgery. AB - BACKGROUND: Nitrous oxide (N(2)O) inhibits methionine synthetase resulting in elevated plasma homocysteine (Hcy) concentration after surgery. In epidemiological studies, hyperhomocysteinaemia is associated with increased risk of cardiovascular disease and dementia. METHODS: Blood samples were obtained to measure plasma folate and Hcy concentrations from two centres participating in a multicentre randomized trial investigating the effects of N(2)O on the outcome after major surgery. The effect of N(2)O and duration of anaesthesia on plasma Hcy, and the relationship between hyperhomocysteinaemia and outcomes were assessed. RESULTS: We enrolled 394 patients. The N(2)O Group had an increase in plasma Hcy concentration after surgery when compared with the N(2)O-free Group: 11.1 (3.8) vs 8.5 (4.0) micromol litre(-1), P<0.0005. Postoperative hyperhomocysteinaemia was associated with an increased risk of major complications: risk ratio (RR) 2.8 (95% CI: 1.4-5.4), P=0.002 and cardiovascular events, RR 5.1 (95% CI: 3.1-8.5), P<0.0005. There was a significant association between duration of anaesthesia and the relative change in plasma Hcy concentration, particularly in the N(2)O Group: r=0.42, P<0.001. CONCLUSIONS: N(2)O increases plasma Hcy concentration; this effect is greater with a longer duration of anaesthesia. Hyperhomocysteinaemia is a risk factor for major postoperative complications. N(2)O-induced increases in plasma Hcy concentration may be a cause of postoperative cardiovascular morbidity. PMID- 18400809 TI - Influence of a continuous prednisolone medication on the time course of neuromuscular block of atracurium in patients with chronic inflammatory bowel disease. AB - BACKGROUND: Corticosteroids interact with neuromuscular blocking agents. However, experimental data are contradictory: enhancement and attenuation of the neuromuscular block has been observed. This study tested the influence of long term medication with prednisolone on atracurium-induced neuromuscular block. METHODS: Sixty patients with chronic inflammatory bowel disease undergoing elective abdominal surgery were investigated. Thirty patients received a long term medication with prednisolone (Group A) and 30 were without corticoid medication (Group B). Additionally, another 30 patients without inflammatory bowel disease and without corticoid medication served as control (Group C). The following parameters of an atracurium-induced neuromuscular block (0.25 mg kg( 1)) were measured: onset time, maximum block, recovery to 25% first twitch height, recovery index (time from 25% until 75% recovery of first twitch), duration to recovery to a train-of-four (TOF) rate of 0.7 and 0.9. RESULTS: The groups did not differ with regard to onset time, maximum block, and recovery index. The duration to 25% twitch height was significantly lower in Group A [18.1 (0-30.7) min] compared with Group B [23.5 (0-36.7) min; P<0.05]. Duration to a TOF rate of 0.7 and 0.9, respectively, were significantly reduced in Group A [36.1 (7.9) and 40.9 (9.0 min)] compared with Group B [47.9 (7.6) and 53.4 (9.2) min; P<0.001]. CONCLUSIONS: Long-term medication with prednisolone resulted in a shorter duration of an atracurium-induced neuromuscular block in patients with Crohn's disease or ulcerative colitis. The presence of the inflammatory bowel disease did not influence the time course of the neuromuscular block. PMID- 18400810 TI - Changes in glomerular filtration rate after cardiac surgery with cardiopulmonary bypass in patients with mild preoperative renal dysfunction. AB - BACKGROUND: Cardiac surgery with cardiopulmonary bypass (CPB) is commonly perceived as a risk factor for decline in renal function, especially in patients with preoperative renal dysfunction. There are few data on the effects of CPB on renal function in patients with mild preoperative renal dysfunction. The purpose of this study was to evaluate renal function in patients with pre-existing mild renal dysfunction undergoing cardiac surgery with CPB. METHODS: In a multicentre study cohort we measured prospectively the glomerular filtration rate (GFR) by radioactive markers both before operation and on the 7th postoperative day in cardiac surgical patients with preoperative serum creatinine >120 micromol l(-1) (n=56). In a subgroup of patients (n=14) in addition to the GFR, the effective renal plasma flow (ERPF) and the filtration fraction (FF) were measured. RESULTS: While preoperative GFR [77.9 (25.5) ml min(-1)] increased to 84.4 (23.7) ml min( 1) (P=0.005) 1 week after surgery, ERPF did not change [295.8 (75.2) ml min(-1) and 295.9 (75.9) ml min(-1), respectively; P=0.8]. In accordance, the FF increased from 0.27 (0.05) (before operation) to 0.30 (0.04) (Day 7, P=0.01). CONCLUSION: Our results oppose the view that cardiac surgery with CPB adversely affects renal function in patients with preoperative mild renal dysfunction and an uncomplicated clinical course. PMID- 18400812 TI - Sevelamer. PMID- 18400813 TI - Do we need another equation to estimate GFR from serum creatinine in renal allograft recipients? PMID- 18400811 TI - Secondary effects of antipsychotics: women at greater risk than men. AB - CONTEXT: The health burden of antipsychotic medication is well known, but the disproportionate effect on women as compared with men is underappreciated. OBJECTIVE: The goal of this article is preventive--to better inform clinicians so that the risks to women and to their offspring can be diminished. METHOD: All PubMed sources in which the search term gender (or sex) was linked to a side effect of antipsychotic medication were reviewed. RESULT: There is general agreement in the literature on women's increased susceptibility to weight gain, diabetes, and specific cardiovascular risks of antipsychotics, with less consensus on malignancy risks and risks to the fetus. Cardiovascular death, to which men are more susceptible than women, is disproportionately increased in women by the use of antipsychotics. Sedating antipsychotics raise the risk of embolic phenomena during pregnancy, and postpartum. Prolactin-elevating drugs suppress gonadal hormone secretion and may enhance autoimmune proclivity. CONCLUSIONS: Clinicians need to be aware of the differential harm that women (and their offspring) can incur from the side effects of antipsychotics. PMID- 18400815 TI - Acute renal failure secondary to oxalosis in a recipient of a simultaneous kidney pancreas transplant: was mycophenolate the cause? PMID- 18400816 TI - The relationship between bone morphogenic protein-7 and peritoneal transport characteristics. AB - BACKGROUND: After prolonged peritoneal dialysis (PD) and exposure to a non physiological dialysis solution, peritoneal mesothelial cells undergo the epithelial-to-mesenchymal transition. In other biological systems, bone morphogenic protein-7 (BMP-7) is a key factor that controls this process. However, the role of BMP-7 in peritoneal physiology has not been studied. METHODS: We studied the peritoneal transport characteristics of 50 consecutive new PD patients at 4 and 52 weeks after PD. Peritoneal permeability will be determined by the standard peritoneal equilibration test (PET). BMP-7 in PD effluent (PDE) at mRNA and protein level at 4 weeks was quantified. RESULTS: At 4 weeks, the mRNA expression of BMP-7 in PDE significantly correlated with peritoneal transport characteristics, including the dialysate-to-plasma creatinine ratio at 4 h (D/P4) (r = 0.422, P = 0.015) and mass transfer area coefficient (MTAC) of creatinine (r = 0.457, P = 0.008). The PDE BMP-7 level by ELISA also had marginal correlation with D/P4 (r = 0.287, P = 0.072) and MTAC creatinine (r = 0.287, P = 0.073), although the result did not reach statistical significance. For the subgroup of patients who remained free of peritonitis, the PDE BMP-7 level by ELISA had significant correlation with the change in D/P4 (r = 0.441, P = 0.017) and MTAC creatinine in 52 weeks (r = 0.415, P = 0.025). The PDE BMP-7 level remained independently associated with the change in peritoneal transport adjusting for age, sex, serum C-reactive protein and PDE transforming growth factor-beta level. In patients who had peritonitis during the study period, the PDE BMP-7 level did not affect the change in peritoneal transport. Conclusion. We find that the peritoneal BMP-7 level correlates with peritoneal transport characteristics, and a high PDE BMP-7 level is associated with a gradual increase in peritoneal transport parameters with time. It remains unclear, however, whether this effect is beneficial, and the therapeutic role of exogenous BMP-7 on peritoneal transport requires a further study. PMID- 18400817 TI - Excess mortality due to interaction between protein-energy wasting, inflammation and cardiovascular disease in chronic dialysis patients. AB - BACKGROUND: Protein-energy wasting (PEW), inflammation and cardiovascular diseases (CVD) clearly contribute to the high mortality in chronic dialysis. Our aim was to examine the presence of additive interaction between these three risk factors in their association with long-term mortality in dialysis patients. METHODS: Patients from a prospective multi-centre cohort study among ESRD patients starting with their first dialysis treatment [the Netherlands Co operative Study on the Adequacy of Dialysis-2 (NECOSAD-II)] with complete data on these risk factors were included (n = 815, age: 59 +/- 15 years, 60% men, 65% HD). Hazard ratios (HR) were calculated for all-cause mortality in 7 years of follow-up. The presence of interaction between the three risk factors was examined, based on additivity of effects. RESULTS: Of all patients, 10% only suffered from PEW (1-5 on the 7-point subjective global assessment), 11% from inflammation (CRP >/=10 mg/L), 14% from CVD and 22% had any combination of two components. Only 6% of the patients had all three risk factors. Patients with either PEW (HR: 1.6, 95% CI: 1.3-2.0), inflammation (1.6, 1.3-2.0) or CVD (1.7, 1.4-2.1) had an increased mortality risk. In patients with all three risk factors, the crude mortality rate of 45/100 person-years was 16 deaths/100 person years higher than expected from the addition of the solo effects of PEW, inflammation and CVD. The relative excess risk due to interaction was 2.9 (95% CI: 0.3-5.4), implying additive interaction. After adjustment for age, sex, treatment modality, primary kidney diseases, diabetes and malignancy the HR for patients with all three risk factors was 4.8 (95% CI: 3.2-7.2). CONCLUSIONS: The concurrent presence of PEW, inflammation and CVD increased the mortality risk strikingly more than expected, implying that PEW interacts with inflammation and CVD in dialysis patients. PMID- 18400818 TI - Haemodialysis in satellite units outside the hospital. PMID- 18400819 TI - Use (or misuse) of vitamin D treatment in CKD and dialysis patients: A recent meta-analysis on vitamin D compounds in chronic kidney disease [1] and an editorial comment [2] accompanying this meta-analysis have already been published. We believe that these papers deserve some comments in the interest of the NDT readership. PMID- 18400820 TI - Application of normobaric hyperoxia therapy for amelioration of haemorrhagic shock-induced acute renal failure. AB - BACKGROUND: Hypoxia resultant from haemorrhagic shock is the primary cause of kidney damage. Application of normobaric hyperoxia therapy (NHT) is an acceptable treatment for acute haemorrhagic shock. We investigated the effect of NHT on amelioration of haemorrhagic shock-induced rat renal failure. METHODS: Twenty four Sprague-Dawley rats were subjected to gradual blood withdrawal/reperfusion, followed by 12-h, 24-h or 48-h NHT. Verification/monitoring of intrarenal hypoxia was performed using Hypoxyprobe-TM-1. Subsequently, cystatin C, urea and creatinine were assessed in serum by a Hitachi autoanalyser, and NO, 3-nitro tyrosine, STAT-8-isoprostane and NF-kB in renal medullae and cortices by specific ELISAs. RESULTS: In rats subjected to haemorrhagic shock, 12- to 48-h NHT significantly reduced intrarenal Hypoxyprobe-TM-1 stained areas and attenuated augmentation of urea, creatinine and cystatin C. Haemorrhagic shock resulted in a 10-fold drop of intrarenal NO availability. 12-h and 24-h, but not 48-h, NHT significantly increased cortical/medullar NO synthesis, the latter, however, not approaching the pre-shock values. Significant shock-induced accumulation of STAT 8-isoprostane and 3-nitro-tyrosine was further exacerbated by NHT. Haemorrhagic shock activated NF-kB in ischaemic tissues, which was not attenuated by NHT. CONCLUSIONS: (1) 12- to 48-h NHT decreased intrarenal hypoxia signs and ameliorated deterioration of renal functions in a rat model of haemorrhagic shock induced renal failure. (2) 12- to 24 h NHT improved bioavailability of NO in cortices/medullae of kidneys recuperating from haemorrhagic shock. (3) If any anti-inflammatory activities were stimulated by NHT, they would not be mediated via the NF-kB pathway. (4) Despite NHT-associated elevation of reactive oxygen species (ROS), early oxygen supply proved mandatory for effective recuperation of ischaemic kidney from detrimental consequences of haemorrhagic shock. PMID- 18400821 TI - Treatment of chronic hepatitis C in haemodialysis patients requires more ribavirin. PMID- 18400827 TI - Discourses on menopause--Part I: Menopause described in texts addressed to Danish women 1996-2004. AB - To understand Danish women's very different ways of interpreting menopausal experiences and the way they construct meaning relating to menopause, it is necessary to include the context in which meaning is constructed as well as the background of cultural attitudes to menopause existing in the Danish society. Using documentary material, the aim of this article was to describe different discourses on menopause in Denmark that present themselves to menopausal women, and to discuss how these discourses may affect women's identity and constitute their scope of action. One hundred and thirty-two pieces of text under the heading or subject of 'menopause' or 'becoming a middle-aged woman', published from 1996 to 2004, were included. All material was addressed to Danish women, and consisted of booklets and informational material, articles from newspapers and magazines and popular science books. Seven different discourses on menopause were identified: the biomedical discourse; the 'eternal youth' discourse; the health promotion discourse; the consumer discourse; the alternative discourse; the feminist/critical discourse; and the existential discourse. The biomedical discourse on menopause was found to be dominant, but was expanded or challenged by other discourses by offering different scopes of action and/or resting on different fundamental values. The discourses constructed and positioned individual women differently; thus, the women's position varied noticeably from one discourse to another. Depending on the discourse drawn upon, the woman's position could be that of a passive patient or that of an empowered woman, capable of making her own choices in relation to her health. PMID- 18400826 TI - The role of the Internet in cancer patients' engagement with complementary and alternative treatments. AB - This article draws on a study of 80 National Health Service cancer patients and their experiences of using the Internet within disease and treatment processes. It focuses on the role the Internet plays in the context of potential or actual engagement with complementary and alternative medicine (CAM). The results depart from previous conceptualizations of the Internet as a major source of CAM knowledge, and second, as a major pathway to patient CAM usage. Moreover, the results highlight significant anxiety as patients attempt to process vast amounts of complex biomedical diagnostic and prognostic information online. For patients attempting to embrace alternative therapeutic models of cancer care, exposure to prognostic data may pose considerable risks to individual well-being and engagement with healing practices. On the basis of these results we problematize social theorizations of the Internet as contributing to such things as: the democratization of knowledge; the deprofessionalization of medicine; and patient empowerment. We emphasize, instead, the potential role of the Internet in reinforcing biomedicine's paradigmatic dominance in cancer care. PMID- 18400828 TI - Discourses on menopause--Part II: How do women talk about menopause? AB - The aim of this article is to describe which of the different available discourses women relate to as revealed in the way they talk about menopause. We use a discourse analytic approach, which implies that meaning is ascribed to things according to how we talk about them. Twenty-four menopausal women from Denmark were interviewed. They were selected to cover a broad spectrum of Danish women with different menopausal experiences and social background factors. Seven previously identified discourses could be found in the interviews, though to varying degrees from woman to woman. Nearly all women used terms from the biomedical sphere like 'a period of decline and decay', even if they did not necessarily agree with this view. Also the existential discourse permeated most of the interviews, especially when the conversation turned to the ageing process, femininity and self-development. The way the menopause was talked about almost became kaleidoscopic when images speedily changed from the decrepit osteoporotic woman or a woman with lack of vitality and sex-appeal to a healthy and strong woman with control over her body and self. Since many women contact doctors in relation to menopause, and since the way doctors talk about menopause is influential, doctors should carefully consider which words and images they use in the counselling. The medical way of perceiving menopause is just one of many, and doctors must be aware that there are other different and partially contradicting discourses at play in society and in the women's universes. PMID- 18400825 TI - Circulatory support with right ventricular assist device and intra-aortic balloon counterpulsation in patient with right ventricle failure after pulmonary embolectomy. AB - Severe pulmonary embolism may lead to acute right ventricular failure despite immediate surgical embolectomy, which is regarded as the treatment of choice after recent CABG surgery. We report a case of a patient with massive pulmonary thromboembolism which resulted in acute right ventricular failure following early surgical embolectomy. Pulmonary embolism developed two days after an elective off pump CABG surgery. We observed severe circulatory collapse which resulted in cardiac arrest and proved refractory to pharmacological treatment after immediate cardiopulmonary resuscitation. Intra-aortic balloon pumping was used in an attempt to improve hemodynamic performance during surgical skin preparation. After the completion of the embolectomy and failure to wean the patient from CPB, upon clinical signs of low cardiac output and akinetic right ventricle, the decision was made to support its function with a centrifugal pump. The substantial improvement of the right ventricular function observed in the next 24 h allowed weaning the patient from right ventricle support. In spite of hemodynamic recovery, the patient remained in a coma on discharge from the cardiac-surgical ICU after 18 days, and died 10 days later from systemic infection. PMID- 18400822 TI - Predictors of change in estimated GFR: a population-based 7-year follow-up from the Tromso study. AB - BACKGROUND: Chronic kidney disease is associated with increased cardiovascular mortality, and even mild impairment of renal function is a cardiovascular risk factor. Several studies have investigated the risk factors for the development of end-stage renal disease, but little is known about predictors of change in renal function in the general population. METHODS: The present study included 2249 men and 2192 women without signs of kidney disease at baseline who were followed for 7 years from 1994 to 1995 in the Tromso Study. Estimated glomerular filtration rate (eGFR) was calculated from the Modification of Diet in Renal Disease study equation. Gender-specific multiple linear regression analyses were used to assess predictors of change in eGFR (DeltaGFR). RESULTS: Change in eGFR, measured in ml/min/1.73 m(2)/year, was associated with systolic blood pressure (SBP) [beta value for a 10-mmHg increase in SBP, men = -0.14, 95% confidence interval (CI) = 0.18 to -0.09; women = -0.07, 95% CI = -0.11 to -0.03] and fibrinogen [beta-value for 1 SD increase in fibrinogen, men (1 SD: 0.85 g/L) = -0.12, 95% CI -0.20 to 0.03; women (1 SD: 0.80) = -0.11, 95% CI -0.20 to -0.02]. High alcohol consumption in men and high physical activity in women predicted an increase in eGFR. Higher albumin/creatinine ratio was associated with a decline in eGFR in men only. CONCLUSIONS: Some risk factors for change in GFR seem to be gender specific but both high SBP and high levels of fibrinogen contribute to a more rapid decline in GFR for both men and women. PMID- 18400829 TI - Structural constraints and opportunities for CAM use and referral by physicians, nurses, and midwives. AB - This article examines the attitudes and reported behaviors of physicians, nurses, and midwives regarding complementary and alternative medicine (CAM), with respect to one key theme that emerged from a broader qualitative study. Of central interest are the structural influences identified by study participants that either act as constraints or opportunities for their professional use of or referral for CAM. In an effort to interpret these results, the analysis is situated within the sociology literature that documents the influences of professional socialization, practice philosophy, regulation, and organizations on professional behavior. These influences are classified as either professional level structures or work/organizational structures. The main conclusion is that future research should pay more attention to such structural dimensions that may be exerting influence on the decisions of providers about whether or not to professionally use or refer for CAM. This article is offered as a conceptual starting point for doing so. PMID- 18400831 TI - A 'little world of your own': stigma, gender and narratives of venereal disease contact tracing. AB - As in other countries, in order to protect the public from venereal disease (syphilis and gonorrhoea), contact tracing in New Zealand has been a public health strategy since the mid-20th century. So far, scholars have predominantly focused on the aspect of control of the cases traced. Based on a rare interview with a female contact tracer, together with a range of archival material, this article aims to expand the scholarship by focusing on the tracer instead of the patient. Using Erving Goffman's original concept of 'courtesy stigma', the article will show that his idea can be nuanced to take into account contact tracers and the ways in which this stigma can be refracted through gender. Working as a tracer had a distinct impact on her life and possibly even her marital status, which were compromised by secrecy, stigma, morality and the demands of public health policies--aspects that were, paradoxically, quite similar to those she traced. The courtesy stigma that contact tracers for venereal disease acquired limited their professional options, as well as isolated them in the non-stigmatized social world. PMID- 18400823 TI - Lack of elevation of urinary albumin excretion among patients with chronic syndromes of inappropriate antidiuresis. AB - BACKGROUND: A recent study has revealed that acute and chronic administration of the vasopressin V2 receptor (V2R) agonist dDAVP induced a marked increase of urinary albumin excretion (UAE) in healthy rats and humans (Bardoux P et al. Nephrol Dial Transplant 2003; 18: 497-506). The occurrence of an elevation of UAE among patients with chronic syndromes of inappropriate antidiuresis has not been reported. METHODS: We looked for the elevation of UAE in 24-h urine samples of the following patients: nine chronic SIADH patients, two patients with acute post operative SIADH, three patients of the same family with nephrogenic syndrome of inappropriate antidiuresis (NSAID) and two patients with hyponatraemia due to surdosage of dDAVP in the setting of central diabetes insipidus. RESULTS: There was no elevation of UAE in our patients (whether they presented with hyponatraemia or not), apart from a patient treated with supra-physiological doses of dDAVP. When she received 80 microg/day of dDAVP, her UAE was 42 mg/day. In this patient, UAE returned to the normal range (21 mg/day) when doses of dDAVP were tapered (20 microg/day). CONCLUSION: The present study shows that chronic V2R stimulation generally does not result in a rise in UAE. The discrepancy between our results and those of the above-mentioned study could be explained by a dose-dependent effect of V2R stimulation on UAE. PMID- 18400832 TI - Medicalization and beyond: the social construction of insomnia and snoring in the news. AB - What role do the media play in the medicalization of sleep problems? This article, based on a British Academy funded project, uses qualitative textual analysis to examine representations of insomnia and snoring in a large representative sample of newspaper articles taken from the UK national press from the mid-1980s to the present day. Constructed as 'common problems' in the population at large, insomnia and snoring we show are differentially located in terms of medicalizing-healthicizing discourses and debates. Our findings also suggest important differences in the gendered construction of these problems and in terms of tabloid and 'broadsheet' newspaper coverage of these issues. Newspaper constructions of sleep, it is concluded, are complex, depending on both the 'problem' and the paper in question. PMID- 18400830 TI - Governing the contagious body: genital herpes, contagion and technologies of the self. AB - Genital herpes is a prevalent sexually transmitted viral infection. While genital herpes is not life-threatening, it can cause physical discomfort and psychosocial difficulties, and may increase the risk of contracting HIV. Given that genital herpes cannot be cured, both the condition itself, and the possibility of passing it on to others, becomes a part of the everyday reality of those individuals diagnosed with genital herpes. In this article we explore the ways in which people with genital herpes attending the Sexually Transmitted Infections (STI) clinic govern their 'contagious bodies'. The discussion draws on the Foucauldian concept of governmentality, and uses Foucault's idea of ethics as a framework to identify the technologies of the self by which individuals with genital herpes govern their own thoughts and behaviours in relation to the contagiousness of the condition. Implications for practice and other ways of thinking about what happens in the STI clinic context are suggested. PMID- 18400834 TI - Role of hypoxia-inducible factor-1alpha in hypoxia-induced expressions of IL-8, MMP-1 and MMP-3 in rheumatoid fibroblast-like synoviocytes. AB - OBJECTIVES: Hypoxia-inducible factor-1alpha (HIF-1alpha) is a master regulator in the cellular response to hypoxic conditions, and rheumatoid synovial tissue is known to exist under hypoxic conditions. Therefore, this study was conducted to determine the contribution of HIF-1alpha to hypoxia-induced MMP and cytokine production in fibroblast-like synoviocytes (FLS). METHODS: RA FLS were transfected with either a plasmid that expresses HIF-1alpha or an empty vector as a control, and then cultured under normoxia (21% O(2)). Also, FLS were transfected with either HIF-1alpha small interfering RNA (siRNA) or control siRNA, and cultured under hypoxic conditions (1% O(2)). Following transfection, the amounts of MMP and cytokine mRNAs and HIF-1alpha protein were examined using real-time RT-PCR and western blotting, respectively. RESULTS: The expression of HIF-1alpha, MMP-1, MMP-3, IL-6 and IL-8 was markedly enhanced in FLS that were cultured under hypoxia. We confirmed that transient transfection of HIF-1alpha overexpressing vector or siRNA had occurred using western blotting, and in vitro studies conducted using FLS transfected with HIF-1alpha overexpression vector showed that they had significantly increased MMP-1, MMP-3 and IL-8 expression levels. Further, hypoxia-induced MMP-3 expression was significantly attenuated by knock-down of HIF-1alpha, whereas hypoxia-induced IL-8 or MMP-1 expression was not significantly repressed by HIF-1alpha siRNA. CONCLUSIONS: Hypoxia-induced MMP 3 expression is exclusively regulated by HIF-1alpha, and hypoxia-induced MMP-1 or IL-8 expression appears to have salvage pathways other than the HIF-1alpha pathway. Together, these data provide new insight regarding the mechanism by which hypoxia participates in joint inflammation and destruction in RA. PMID- 18400842 TI - Turner syndrome and clinical treatment. AB - BACKGROUND: Turner syndrome (TS) is a genetic disorder associated with abnormalities of the X chromosome, occurring in about 50 per 100,000 liveborn girls. TS is usually associated with reduced adult height, gonadal dysgenesis and thus insufficient circulating levels of female sex steroids leading to premature ovarian failure and infertility. The average intellectual performance is within the normal range. New insight into genetics, epidemiology, cardiology, endocrinology and metabolism from a number of recent studies will be included in this review. SOURCES OF DATA: For this review we concentrated on all papers published on TS with special emphasis on the most recent literature. Also papers relating to cardiology, especially aortic dissection, paediatrics and the effects of estradiol in other conditions were considered. The main source was PubMed and the major endocrinology and cardiology journals. AREAS OF AGREEMENT: Treatment with growth hormone (GH) during childhood and adolescence allows a considerable gain in adult height. SHOX deficiency explains some of the phenotypic characteristics in TS, principally short stature. Puberty has to be induced in most cases, and female sex hormone replacement therapy (HRT) is given during adult years. Morbidity and mortality is increased, especially due to the risk of dissection of the aorta and other cardiovascular (CV) diseases, as well as the risk of type 2 diabetes, osteoporosis and thyroid disease. AREAS OF CONTROVERSY: The proper dose of HRT with female sex steroids has not been established, and, likewise, benefits and/or drawbacks from HRT have not been thoroughly evaluated. In most countries it seems that the transition period from paediatric to adult care is especially vulnerable and the proper framework for transition has not been established. Today, most treatment recommendations are based on expert opinion and are unfortunately not evidence based, although more areas, such as GH treatment for increasing height, are well founded. GROWING POINTS: The description of adult life with TS has been broadened and medical, social and psychological aspects are being added at a compelling pace. AREAS TIMELY FOR DEVELOPING RESEARCH: Proper care during adulthood should be studied, since most morbidity potentially is amenable to proper care. Especially, interventional strategy and follow-up with respect to congenital CV malformations, as well as secondary CV disease, have to be developed and new treatment algorithms have to be studied. In summary, TS is a condition associated with a number of diseases and conditions, which need the attention of a multi-disciplinary team. PMID- 18400836 TI - Frequency and duration of clinical remission in patients with peripheral psoriatic arthritis requiring second-line drugs. AB - OBJECTIVE: To evaluate the frequency and duration of clinical remission in patients with PsA. METHODS: All consecutive new outpatients with peripheral PsA requiring second-line drugs and RA observed between January 2000 and December 2005 were included in a prospective, case-control study. Primary end point was to assess the frequency of remission in peripheral PsA compared with RA. Secondary end points were to compare the duration of clinical remission during treatment and after therapy interruption, ACR 20, 50, 70 response rates and to detect any remission predictor at diagnosis. Treatment regimen was standardized in both groups. From January 2003 to December 2005, therapy was suspended in PsA patients and controls if achieving remission. RESULTS: One or more episodes of remission occurred in 57/236 (24.1%) PsA patients and in 20/268 (7.5%) controls (P < 0.001). The mean duration of remission was of 13 +/- 9.4 months in PsA patients and 4 +/- 3.7 in controls (P > 0.001). Remission episodes were more frequent in PsA patients treated with anti-TNF compared with those receiving traditional DMARDs (P > 0.001), with no differences regarding the duration. After therapy interruption, the remission duration was 12 +/- 2.4 months in PsA and 3 +/- 1.5 in RA (P < 0.001). No remission predictor at diagnosis resulted by multivariate analysis. CONCLUSION: Remission is possible in up to 24% of patients with peripheral PsA. It is significantly more frequent, but not longer, in patients receiving anti-TNF drugs compared with those treated with traditional DMARDs. Patients remain in remission for a long period after therapy interruption, thus suggesting an intermittent therapeutic strategy. PMID- 18400835 TI - Functional epitope of muscarinic type 3 receptor which interacts with autoantibodies from Sjogren's syndrome patients. AB - OBJECTIVES: Recently, autoantibodies directed against muscarinic type 3 receptor (M3R) have been reported in patients with primary SS. However, the precise epitope(s) of the M3R that interacts with SS autoantibodies remains unclear. The aim of this study was to identify the functional epitope of M3R which interacts with SS immunoglobulin G (IgG). METHODS: Purified IgGs were obtained from the sera of seven SS patients (six primary and one secondary SS) and two normal persons. We examined whether SS IgG inhibits M3R function and identified the epitope using six synthetic peptides covering all the extracellular domains of M3R by microspectrofluorimetry and surface plasmon resonance-based optical biosensor system (BIAcore system). RESULTS: A volume of 0.5 mg/ml SS IgG inhibited carbachol (CCh)-induced [Ca(2+)](i) transient (CICT) in human submandibular gland (HSG) cells. However, co-incubation of SS IgG with the 6th peptide (514-527 amino acid region) corresponding to the third extracellular loop of M3R, recovered CICT. The result was further confirmed by BIAcore analysis. We found that the 6th peptide interacts with IgGs from three primary SS patients in a concentration-dependent manner. The synthetic peptide which consists of amino acids 228-237 corresponding to the COOH-terminus of the second extracellular loop of M3R also bound to SS IgG. However, normal IgGs did not interact with the 6th peptide. CONCLUSIONS: The results suggest that the third extracellular loop of M3R represents a functional epitope bound by SS IgG, and thereby partly inhibits M3R function. PMID- 18400833 TI - Cardiovascular risk profile of patients with psoriatic arthritis compared to controls--the role of inflammation. AB - OBJECTIVE: To examine the distribution of traditional and novel risk factors of cardiovascular disease (CVD) in patients with PsA compared with healthy controls. METHODS: We compared risk factors for CVD between 102 consecutive PsA patients and 82 controls, adjusting for BMI. We also assessed the role of inflammation on the CVD risk factor by using a BMI and high-sensitivity CRP (hsCRP)-adjusted model. RESULTS: The BMI of PsA patients were significantly higher than healthy controls. After adjusting for the BMI, PsA patients still have a higher prevalence of diabetes mellitus (DM) [odds ratio (OR) 9.27, 95% CI 2.09, 41.09) and hypertension (OR 3.37, 95% CI 1.68, 6.72), but a lower prevalence of low high density lipoprotein (HDL) cholesterol (OR 0.16, 95% CI 0.07, 0.41). PsA patients have significantly increased systolic and diastolic blood pressures, insulin resistance and inflammatory markers (hsCRP and white cell count) compared to controls. PsA patients have higher HDL cholesterol and apolipoprotein (Apo) A1 levels; and lower total cholesterol (TC) and low density lipoprotein cholesterol levels; and a lower TC/HDL ratio. However, the Apo B level (P < 0.05), and the Apo B/Apo A1 ratio (P = 0.07) were higher in PsA patients. Further adjustment for hsCRP level rendered the differences in the prevalence of hypertension and DM; the TC, and sugar levels; and white cell count non-significant between the two groups; while the differences in other parameters remained significant. CONCLUSION: These data support the hypothesis that PsA may be associated with obesity, hypertension, dyslipidaemia and insulin resistance because of the shared inflammatory pathway. PMID- 18400839 TI - Pre-natal exposure to paracetamol and risk of wheezing and asthma in children: a birth cohort study. AB - BACKGROUND: Paracetamol use has been associated with increased prevalence of asthma in children and adults, and one study reported an association between pre natal exposure to paracetamol and asthma in early childhood. METHODS: To examine if pre-natal exposure to paracetamol is associated with the risk of asthma or wheezing in early childhood, we selected 66 445 women from the Danish National Birth Cohort for whom we had information on paracetamol use during pregnancy and who participated in an interview when their children were 18-months-old and 12 733 women whose children had reached the age of 7 and estimated the prevalence of physician-diagnosed asthma and wheezing at the ages of 18 months and 7 years. We also linked our population to the Danish National Hospital Registry to record all hospitalizations due to asthma up to age of 18 months. RESULTS: Paracetamol use during any time of pregnancy was associated with a small but statistically significant increased risk of physician-diagnosed asthma or bronchitis among children at 18 months [relative risk (RR) = 1.17, 1.13-1.23)], hospitalizations due to asthma up to 18 months (hazard ratio = 1.24, 1.11-1.38) and physician diagnosed asthma at 7 years (RR = 1.15, 1.02-1.29). The highest risks were observed for paracetamol use during the first trimester of pregnancy and persistent wheezing (wheezing at both 18 months and 7 years) (RR = 1.45, 1.13 1.85). CONCLUSION: Paracetamol use during pregnancy was associated with an increased risk of asthma and wheezing in childhood. If this association is causal, we may need to revisit the clinical practice on use of paracetamol during pregnancy. PMID- 18400841 TI - Metabolic correlates of subthalamic nucleus activity in Parkinson's disease. AB - Overactivity of subthalamic nucleus (STN) neurons is a consistent feature of Parkinson's disease (PD) and is a target of therapy for this disorder. However, the relationship of STN firing rate to regional brain function is not known. We scanned 17 PD patients with (18)F-fluorodeoxyglucose (FDG) PET to measure resting glucose metabolism before the implantation of STN deep brain stimulation electrodes. Spontaneous STN firing rates were recorded during surgery and correlated with preoperative regional glucose metabolism on a voxel-by-voxel basis. We also examined the relationship between firing rate and the activity of metabolic brain networks associated with the motor and cognitive manifestations of the disease. Mean firing rates were 47.2 +/- 6.1 and 48.7 +/- 8.5 Hz for the left and right hemispheres, respectively. These measures correlated (P < 0.007) with glucose metabolism in the putamen and globus pallidus, which receive projections from this structure. Significant correlations (P < 0.0005) were also evident in the primary motor (BA4) and dorsolateral prefrontal (BA46/10) cortical areas. The activity of both the motor (P < 0.0001) and the cognitive (P < 0.006) PD-related metabolic networks was elevated in these patients. STN firing rates correlated with the activity of the former (P < 0.007) but not the latter network (P = 0.39). The findings suggest that the functional pathways associated with motor disability in PD are linked to the STN firing rate. These pathways are likely to mediate the clinical benefit that is seen following targeted STN interventions for this disease. PMID- 18400838 TI - Validity of the Pediatric Quality Of Life Inventory for youth with sickle cell disease. AB - OBJECTIVE: Evaluate the validity of the Pediatric Quality of Life Inventory (PedsQL) for sickle cell disease (SCD). METHODS: Sixty-eight parent-child dyads (children 5-18 years) completed the PedsQL. Medical record review assessed history of specific morbidities. RESULTS: Internal consistency of the scales varied. The strongest reliability was for parent proxy-report for specific domains or for global functioning scores with either informant. Modest internal consistency was found for specific domains with child informants, particularly for younger children. Moderate convergent validity was found between informants. History of neurologic problems or major pain episodes indicated criterion validity for specific scales. CONCLUSIONS: The PedsQL appears to validly assess quality of life in youth with SCD. Domain-specific measurement of quality of life was limited by (a) low reliability for youth-report and (b) lack of discriminant validity. Choice of informant may be important when evaluating quality of life effects from pain or neurologic problems in SCD. PMID- 18400840 TI - Intrauterine devices and intrauterine systems. AB - From a public health viewpoint, the intrauterine device (IUD) is the most widely used contraceptive method in the world. Prevalence rates range among countries from 2 to 80% of contraceptive users. During 5 years of IUD use, pregnancy occurs in less than 2 per 100 insertions. Bleeding and pain are the most common reasons for removal rates of 10% in the first year and up to 50% within 5 years. The contraceptive effects of IUDs may be due to a sterile inflammatory reaction in the endometrial cavity which interferes with sperm function, so that fertilization is less likely to occur. IUDs also interfere with implantation but the extent to which this contributes to their contraceptive action is unknown. In nulliparous women rates of expulsion and removal for bleeding and/or pain are higher than in parous women. Effective use of IUDs for up to 10 years has the same pregnancy rate as tubal interruption. Thus, the IUD may be an alternative to female sterilization, especially in younger women who are more likely to experience regret after sterilization. The levonorgestrel intrauterine system (LNG-IUS) reduces bleeding and dysmenorrhoea, provides superior effectiveness to copper IUDs, and may be a useful treatment for endometriosis or an alternative to hysterectomy for menorrhagia. PMID- 18400837 TI - Value of serum cartilage oligomeric matrix protein as a prognostic marker of large-joint damage in rheumatoid arthritis--data from the RAPIT study. AB - OBJECTIVE: To investigate the utility of serum COMP level measurements as a predictor of future damage of the weight-bearing (large) joints in RA patients participating in intensive exercise. METHODS: Data of the 281 completers of a 2 yr randomized controlled trial (Rheumatoid Arthritis Patients In Training; RAPIT) comparing the effects of usual care physical therapy with high-intensity weight bearing exercises were analysed. The primary outcome variable was defined as the change in radiological joint damage (Larsen score) of the large joints. Potential predictors of outcome were defined: baseline and change in serum level of COMP after 3 months, baseline radiological damage of the large and small joints, number of months on glucocorticoids, change in disease activity and in physical capacity (aerobic fitness and muscle strength) after 2 yrs, and participation in the exercise group. RESULTS: In cross-sectional evaluation of baseline data, we found strong association between the high serum COMP level and current damage of the large joints. Serum COMP level at baseline, however, was not associated with an increased rate of radiological joint damage after 2 yrs of follow-up. Furthermore, neither interaction between baseline COMP level and participation in exercises, nor change in COMP level after 3 months of exercising were associated with future damage of the large joints. CONCLUSION: Neither baseline serum COMP level nor its individual change after 3 months from start of intensive exercise predict longitudinal progression of damage of the large joints in this population. PMID- 18400843 TI - Transglutaminase-catalyzed transamidation: a novel mechanism for Rac1 activation by 5-hydroxytryptamine2A receptor stimulation. AB - Transglutaminase (TGase)-induced activation of small G proteins via 5 hydroxytryptamine (HT)(2A) receptor signaling leads to platelet aggregation (Cell 115:851-862, 2003). We hypothesize that stimulation of 5-HT(2A) receptors in neurons activates TGase, resulting in transamidation of serotonin to a small G protein, Rac1, thereby constitutively activating Rac1. Using immunoprecipitation and immunoblotting, we show that, in rat cortical cell line A1A1v, serotonin increases TGase-catalyzed transamidation of Rac1. This transamidation occurs in both undifferentiated and differentiated cells. Treatment with a 5-HT(2A/2C) receptor agonist 2,5-dimethoxy-4-iodoamphetamine, but not the 5-HT(1A) receptor agonist 5-hydroxy-2-dipropylamino tetralin, increases transamidation of Rac1 by TGase. In A1A1v cells, 5-HT(2A) receptors mediate the transamidation reaction because expression of 5-HT(2C) receptors was not detectable and the selective 5 HT(2A) receptor antagonist blocked transamidation. Time course studies demonstrate that transamidation of Rac1 is significantly elevated after 5 and 15 min of serotonin treatment, but returns it to control levels after 30 min. The activity of Rac1 is also transiently increased following serotonin stimulation. Inhibition of TGase by cystamine or small interfering RNA reduces TGase modification of Rac1, and cystamine also prevents Rac1 activation. Serotonin itself is bound to Rac1 by TGase following 5-HT(2A) receptor stimulation as demonstrated by coimmunoprecipitation experiments and a dose-dependent decrease of serotonin-associated Rac1 by cystamine. These data support the hypothesis that Rac1 activity is transiently increased due to TGase-catalyzed transamidation of serotonin to Rac1 via stimulation of 5-HT(2A) receptors. Activation of Rac1 via TGase is a novel effector and second messenger of the 5-HT(2A) receptor-signaling cascade in neurons. PMID- 18400845 TI - Silencing of rotavirus NSP4 or VP7 expression reduces alterations in Ca2+ homeostasis induced by infection of cultured cells. AB - Rotavirus infection of cells in culture induces major changes in Ca(2+) homeostasis. These changes include increases in plasma membrane Ca(2+) permeability, cytosolic Ca(2+) concentration, and total cell Ca(2+) content and a reduction in the amount of Ca(2+) released from intracellular pools sensitive to agonists. Various lines of evidence suggest that the nonstructural glycoprotein NSP4 and possibly the major outer capsid glycoprotein VP7 are responsible for these effects. In order to evaluate the functional roles of NSP4 and other rotavirus proteins in the changes in Ca(2+) homeostasis observed in infected cells, the expressions of NSP4, VP7, and VP4 were silenced using the short interfering RNA (siRNA) technique. The transfection of specific siRNAs resulted in a strong and specific reduction of the expression of NSP4, VP7, and VP4 and decreased the yield of new viral progeny by more than 90%. Using fura-2 loaded cells, we observed that knocking down the expression of NSP4 totally prevented the increase in Ca(2+) permeability of the plasma membrane and cytosolic Ca(2+) concentration measured in infected cells. A reduction in the levels of VP7 expression partially reduced the effect of infection on plasma membrane Ca(2+) permeability and Ca(2+) pools released by agonist (ATP). In addition, the increase of total Ca(2+) content (as measured by (45)Ca(2+) uptake) observed in infected cells was reduced to the levels in mock-infected cells when NSP4 and VP7 were silenced. Finally, when the expression of VP4 was silenced, none of the disturbances of Ca(2+) homeostasis caused by rotaviruses in infected cells were affected. These data altogether indicate that NSP4 is the main protein responsible for the changes in Ca(2+) homeostasis observed in rotavirus-infected cultured cells. Nevertheless, VP7 may contribute to these effects. PMID- 18400846 TI - Functional and structural dynamics of hepadnavirus reverse transcriptase during protein-primed initiation of reverse transcription: effects of metal ions. AB - Reverse transcription in hepadnaviruses is primed by the viral reverse transcriptase (RT) (protein priming) and requires the interaction between the RT and a specific viral RNA template termed epsilon. Protein priming is resistant to a number of RT inhibitors that can block subsequent viral DNA elongation and likely requires a distinct "priming" conformation. Furthermore, protein priming may consist of two distinct stages, i.e., the attachment of the first deoxynucleotide to RT (initiation) and the subsequent addition of 2 or 3 deoxynucleotides (polymerization). In particular, a truncated duck hepatitis B virus RT (MiniRT2) is competent in initiation but defective in polymerization when tested in the presence of Mg(2+). Given the known effects of metal ions on the activities of various DNA and RNA polymerases, we tested if metal ions could affect hepadnavirus RT priming. We report here that Mn(2+), in comparison with Mg(2+), showed dramatic effects on the priming activity of MiniRT2 as well as the full-length RT. First and foremost, MiniRT2 exhibited full polymerization activity in the presence of Mn(2+), indicating that MiniRT2 contains all sequences essential for polymerization but is unable to transition from initiation to polymerization with Mg(2+). Second, the initiation activities of MiniRT2 and the full-length RT were much stronger with Mn(2+). Third, the nucleotide and template specificities during protein priming were decreased in the presence of Mn(2+). Fourth, polymerization was sensitive to inhibition by a pyrophosphate analog in the presence of Mn(2+) but not in the presence of Mg(2+). Finally, limited proteolysis provided direct evidence that the priming active MiniRT2 adopted distinct conformations depending on the presence of Mn(2+) versus that of Mg(2+) and that the transition from initiation to polymerization was accompanied by RT conformational change. PMID- 18400848 TI - Mutagenesis of the active-site cysteine in the ubiquitin-specific protease contained in large tegument protein pUL36 of pseudorabies virus impairs viral replication in vitro and neuroinvasion in vivo. AB - Herpesviruses specify a ubiquitin-specific protease activity located within their largest tegument protein. Although its biological role is still largely unclear, mutation within the active site abolished deubiquitinating (DUB) activity and decreased virus replication in vitro and in vivo. To further elucidate the role of DUB activity for herpesvirus replication, the conserved active-site cysteine at amino acid position 26 within pUL36 of Pseudorabies virus (PrV) (Suid herpesvirus 1), a neurotropic alphaherpesvirus, was mutated to serine. Whereas one-step growth kinetics of the resulting mutant virus PrV-UL36(C(26)S) were moderately reduced, plaque size was decreased to 62% of that of the wild-type virus. Ultrastructural analysis revealed large accumulations of unenveloped nucleocapsids in the cytoplasm, but incorporation of the tegument protein pUL37 was not abolished. After intranasal infection with PrV-UL36(C(26)S) mice showed survival times two times longer than those of mice infected with wild-type or rescued virus. Thus, the DUB activity is important for PrV replication in vitro and for neuroinvasion in mice. PMID- 18400844 TI - The FUSE binding protein is a cellular factor required for efficient replication of hepatitis C virus. AB - Hepatitis C virus (HCV) infection is the leading cause of liver cirrhosis and hepatocellular carcinoma and one of the primary indications for liver transplantation. The molecular mechanisms underlying the actions of host factors in HCV replication remain poorly defined. FUSE (far upstream element of the c-myc proto-oncogene) binding protein (FBP) is a cellular factor that we have identified as a binder of HCV 3' nontranslated region (3'NTR). Mapping of the binding site showed that FBP specifically interacts with the poly(U) tract within the poly(U/UC) region of the 3'NTR. Silencing of FBP expression by small interfering RNA in cells carrying HCV subgenomic replicons severely reduced viral replication, while overexpression of FBP significantly enhanced viral replication. We confirmed these observations by an in vitro HCV replication assay in the cell-free replicative lysate, which suggested that there is a direct correlation between the cellular FBP level and HCV replication. FBP immunoprecipitation coprecipitated HCV nonstructural protein 5A (NS5A), indicating that FBP interacts with HCV NS5A, which is known to function as a link between HCV translation and replication. Although FBP is mainly localized in the nucleus, we found that in MH14 cells a significant level of this protein is colocalized with NS5A in the cytosol, a site of HCV replication. While the mechanism of FBP involvement in HCV replication is yet to be delineated, our findings suggest that it may be an important regulatory component that is essential for efficient replication of HCV. PMID- 18400847 TI - Functions of the ORF9-to-ORF12 gene cluster in varicella-zoster virus replication and in the pathogenesis of skin infection. AB - The gene cluster composed of varicella-zoster virus (VZV) open reading frame 9 (ORF9) to ORF12 encodes four putative tegument proteins and is highly conserved in most alphaherpesviruses. In these experiments, the genes within this cluster were deleted from the VZV parent Oka (POKA) individually or in combination, and the consequences for VZV replication were evaluated with cultured cells in vitro and with human skin xenografts in SCID mice in vivo. As has been reported for ORF10, ORF11 and ORF12 were dispensable for VZV replication in melanoma and human embryonic fibroblast cells. In contrast, deletion of ORF9 was incompatible with the recovery of infectious virus. ORF9 localized to the virion tegument and formed complexes with glycoprotein E, which is an essential protein, in VZV infected cells. Recombinants lacking ORF10 and ORF11 (POKADelta10/11), ORF11 and ORF12 (POKADelta11/12), or ORF10, ORF11 and ORF12 (POKADelta10/11/12) were viable in cultured cells. Their growth kinetics did not differ from those of POKA, and nucleocapsid formation and virion assembly were not disrupted. In addition, these deletion mutants showed no differences compared to POKA in infectivity levels for primary human tonsil T cells. Deletion of ORF12 had no effect on skin infection, whereas replication of POKADelta11, POKADelta10/11, and POKADelta11/12 was severely reduced, and no virus was recovered from skin xenografts inoculated with POKADelta10/11/12. These results indicate that with the exception of ORF9, the individual genes within the ORF9-to-ORF12 gene cluster are dispensable and can be deleted simultaneously without any apparent effect on VZV replication in vitro but that the ORF10-to-ORF12 cluster is essential for VZV virulence in skin in vivo. PMID- 18400849 TI - Definition of a conserved immunodominant domain on hepatitis C virus E2 glycoprotein by neutralizing human monoclonal antibodies. AB - Development of a successful hepatitis C virus (HCV) vaccine requires the definition of neutralization epitopes that are conserved among different HCV genotypes. Five human monoclonal antibodies (HMAbs) are described that cross compete with other antibodies to a cluster of overlapping epitopes, previously designated domain B. Each HMAb broadly neutralizes retroviral pseudotype particles expressing HCV E1 and E2 glycoproteins, as well as the infectious chimeric genotype 1a and genotype 2a viruses. Alanine substitutions of residues within a region of E2 involved in binding to CD81 showed that critical E2 contact residues involved in the binding of representative antibodies are identical to those involved in the binding of E2 to CD81. PMID- 18400851 TI - E protein domain III determinants of yellow fever virus 17D vaccine strain enhance binding to glycosaminoglycans, impede virus spread, and attenuate virulence. AB - The yellow fever virus (YFV) 17D strain is one of the most effective live vaccines for human use, but the in vivo mechanisms for virulence attenuation of the vaccine and the corresponding molecular determinants remain elusive. The vaccine differs phenotypically from wild-type YFV by the loss of viscerotropism, despite replicative fitness in cell culture, and genetically by 20 amino acid changes predominantly located in the envelope (E) protein. We show that three residues in E protein domain III inhibit spread of 17D in extraneural tissues and attenuate virulence in type I/II interferon-deficient mice. One of these residues (Arg380) is a dominant glycosaminoglycan-binding determinant, which mainly accounts for more rapid in vivo clearance of 17D from the bloodstream in comparison to 17D-derived variants with wild-type-like E protein. While other mutations will account for loss of neurotropism and phenotypic stability, the described impact of E protein domain III changes on virus dissemination and virulence is the first rational explanation for the safety of the 17D vaccine in humans. PMID- 18400850 TI - Characterization of neutralizing antibody responses elicited by clade A envelope immunogens derived from early transmitted viruses. AB - The vast majority of studies with candidate immunogens based on the human immunodeficiency virus envelope (Env) have been conducted with Env proteins derived from clade B viruses isolated during chronic infection. Whether non-clade B Env protein immunogens will elicit antibodies with epitope specificities that are similar to those of antibodies elicited by clade B Envs and whether the antibodies elicited by Envs derived from early transmitted viruses will be similar to those elicited by Envs derived from viruses isolated during chronic infection are currently unknown. Here we performed immunizations with four clade A Envs, cloned directly from the peripheral blood of infected individuals during acute infection, which differed in lengths and extents of glycosylation. The antibody responses elicited by these four Envs were compared to each other and to those elicited by a well-characterized clade B Env immunogen derived from the SF162 virus, which was isolated during chronic infection. Only one clade A Env, the one with the fewer glycosylation sites, elicited homologous neutralizing antibodies (NAbs); these did not target the V1, V2, or V3 regions. In contrast, all four clade A Envs elicited anti-V3 NAbs against "easy-to-neutralize" clade B and clade A isolates, irrespective of the variable region length and extent of glycosylation of the Env used as an immunogen. These anti-V3 NAbs did not access their epitopes on homologous and heterologous clade A, or B, neutralization resistant viruses. The length and extent of glycosylation of the variable regions on the clade A Env immunogens tested did not affect the breadth of the elicited NAbs. Our data also indicate that the development of cross-reactive NAbs against clade A viruses faces similar hurdles to the development of cross-reactive anti clade B NAbs. PMID- 18400853 TI - Replication in a superficial epithelial cell niche explains the lack of pathogenicity of primate foamy virus infections. AB - Foamy viruses (FVs) are ancient retroviruses that are ubiquitous in nonhuman primates (NHPs). While FVs share many features with pathogenic retroviruses, such as human immunodeficiency virus, FV infections of their primate hosts have no apparent pathological consequences. Paradoxically, FV infections of many cell types in vitro are rapidly cytopathic. Previous work has shown that low levels of proviral DNA are found in most tissues of naturally infected rhesus macaques, but these proviruses are primarily latent. In contrast, viral RNA, indicative of viral replication, is restricted to tissues of the oral mucosa, where it is abundant. Here, we perform in situ hybridization on tissues from rhesus macaques naturally infected with simian FV (SFV). We show that superficial differentiated epithelial cells of the oral mucosa, many of which appear to be shedding from the tissue, are the major cell type in which SFV replicates. Thus, the innocuous nature of SFV infection can be explained by replication that is limited to differentiated superficial cells that are short-lived and shed into saliva. This finding can also explain the highly efficient transmission of FVs among NHPs. PMID- 18400852 TI - Poliovirus 2A(Pro) increases viral mRNA and polysome stability coordinately in time with cleavage of eIF4G. AB - Poliovirus (PV) 2A protease (2A(Pro)) cleaves eukaryotic initiation factors 4GI and 4GII (eIF4GI and eIF4GII) within virus-infected cells, effectively halting cap-dependent mRNA translation. PV mRNA, which does not possess a 5' cap, is translated via cap-independent mechanisms within viral protease-modified messenger ribonucleoprotein (mRNP) complexes. In this study, we determined that 2A(Pro) activity was required for viral polysome formation and stability. 2A(Pro) cleaved eIF4GI and eIF4GII as PV polysomes assembled. A 2A(Cys109Ser) (2A(Pro) with a Cys109Ser mutation) protease active site mutation that prevented cleavage of eIF4G coordinately inhibited the de novo formation of viral polysomes, the stability of viral polysomes, and the stability of PV mRNA within polysomes. 2A(Cys109Ser)-associated defects in PV mRNA and polysome stability correlated with defects in PV mRNA translation. 3C(Pro) activity was not required for viral polysome formation or stability. 2A(Pro)-mediated cleavage of eIF4G along with poly(rC) binding protein binding to the 5' terminus of uncapped PV mRNA appear to be concerted mechanisms that allow PV mRNA to form mRNP complexes that evade cellular mRNA degradation machinery. PMID- 18400854 TI - Poly(rC) binding proteins and the 5' cloverleaf of uncapped poliovirus mRNA function during de novo assembly of polysomes. AB - Poliovirus (PV) mRNA is unusual because it possesses a 5'-terminal monophosphate rather than a 5'-terminal cap. Uncapped mRNAs are typically degraded by the 5' exonuclease XRN1. A 5'-terminal cloverleaf RNA structure interacts with poly(rC) binding proteins (PCBPs) to protect uncapped PV mRNA from 5' exonuclease (K. E. Murray, A. W. Roberts, and D. J. Barton, RNA 7:1126-1141, 2001). In this study, we examined de novo polysome formation using HeLa cell-free translation replication reactions. PV mRNA formed polysomes coordinate with the time needed for ribosomes to traverse the viral open reading frame (ORF). Nascent PV polypeptides cofractionated with viral polysomes, while mature PV proteins were released from the polysomes. Alterations in the size of the PV ORF correlated with alterations in the size of polysomes with ribosomes present every 250 to 500 nucleotides of the ORF. Eukaryotic initiation factor 4GI (eIF4GI) was cleaved rapidly as viral polysomes assembled and the COOH-terminal portion of eIF4GI cofractionated with viral polysomes. Poly(A) binding protein, along with PCBP 1 and 2, also cofractionated with viral polysomes. A C24A mutation that inhibits PCBP-5'-terminal cloverleaf RNA interactions inhibited the formation and stability of nascent PV polysomes. Kinetic analyses indicated that the PCBP-5' cloverleaf RNA interaction was necessary to protect PV mRNA from 5' exonuclease immediately as ribosomes initially traversed the viral ORF, before viral proteins could alter translation factors within nascent polysomes or contribute to ribonucleoprotein complexes at the termini of the viral mRNA. PMID- 18400855 TI - A subgenomic segment of Theiler's murine encephalomyelitis virus RNA causes demyelination. AB - The DA strain of Theiler's murine encephalomyelitis virus (TMEV) causes a persistent central nervous system (CNS) infection of mice with a restricted virus gene expression and induces an inflammatory demyelinating disease that is thought to be immune mediated and a model of multiple sclerosis (MS). The relative contribution of virus vis-a-vis the immune system in the pathogenesis of DA induced white matter disease remains unclear, as is also true in MS. To clarify the pathogenesis of DA-induced demyelination, we used Cre/loxP technology to generate a transgenic mouse that has tamoxifen (Tm)-inducible expression of a subgenomic segment of DA RNA in oligodendrocytes and Schwann cells. Tm-treated young transgenic mice developed progressive weakness leading to death, with abnormalities of oligodendrocytes and Schwann cells and demyelination, but without inflammation, demonstrating that DA virus can play a direct pathogenic role in demyelination. Tm treatment of mice at a later age resulted in milder disease, with evidence of peripheral nerve remyelination and focal fur depigmentation; surviving weak mice had persistent expression of the recombined transgene in the CNS, suggesting that the DA subgenomic segment can cause cellular dysfunction but not death, possibly similar to the situation seen during DA virus persistence. These studies demonstrate that DA RNA or a DA protein(s) is toxic to myelin-synthesizing cells. This Cre/loxP transgenic system allows for spatially and temporally controlled expression of the viral transgene and is valuable for clarifying nonimmune (and immune) mechanisms of demyelination induced by TMEV as well as other viruses. PMID- 18400856 TI - Critical role of conserved hydrophobic residues within the major homology region in mature retroviral capsid assembly. AB - During retroviral maturation, the CA protein undergoes dramatic structural changes and establishes unique intermolecular interfaces in the mature capsid shell that are different from those that existed in the immature precursor. The most conserved region of CA, the major homology region (MHR), has been implicated in both immature and mature assembly, although the precise contribution of the MHR residues to each event has been largely undefined. To test the roles of specific MHR residues in mature capsid assembly, an in vitro system was developed that allowed for the first-time formation of Rous sarcoma virus CA into structures resembling authentic capsids. The ability of CA to assemble organized structures was destroyed by substitutions of two conserved hydrophobic MHR residues and restored by second-site suppressors, demonstrating that these MHR residues are required for the proper assembly of mature capsids in addition to any role that these amino acids may play in immature particle assembly. The defect caused by the MHR mutations was identified as an early step in the capsid assembly process. The results provide strong evidence for a model in which the hydrophobic residues of the MHR control a conformational reorganization of CA that is needed to initiate capsid assembly and suggest that the formation of an interdomain interaction occurs early during maturation. PMID- 18400857 TI - The LEM domain proteins emerin and LAP2alpha are dispensable for human immunodeficiency virus type 1 and murine leukemia virus infections. AB - The human nuclear envelope proteins emerin and lamina-associated polypeptide 2alpha (LAP2alpha) have been proposed to aid in the early replication steps of human immunodeficiency virus type 1 (HIV-1) and murine leukemia virus (MLV). However, whether these factors are essential for HIV-1 or MLV infection has been questioned. Prior studies in which conflicting results were obtained were highly dependent on RNA interference-mediated gene silencing. To shed light on these contradictory results, we examined whether HIV-1 or MLV could infect primary cells from mice deficient for emerin, LAP2alpha, or both emerin and LAP2alpha. We observed HIV-1 and MLV infectivity in mouse embryonic fibroblasts (MEFs) from emerin knockout, LAP2alpha knockout, or emerin and LAP2alpha double knockout mice to be comparable in infectivity to wild-type littermate-derived MEFs, indicating that both emerin and LAP2alpha were dispensable for HIV-1 and MLV infection of dividing, primary mouse cells. Because emerin has been suggested to be important for infection of human macrophages by HIV-1, we also examined HIV-1 transduction of macrophages from wild-type mice or knockout mice, but again we did not observe a difference in susceptibility. These findings prompted us to reexamine the role of human emerin in supporting HIV-1 and MLV infection. Notably, both viruses efficiently infected human cells expressing high levels of dominant-negative emerin. We thus conclude that emerin and LAP2alpha are not required for the early replication of HIV-1 and MLV in mouse or human cells. PMID- 18400858 TI - Ninety-nine is not enough: molecular characterization of inhibitor-resistant human immunodeficiency virus type 1 protease mutants with insertions in the flap region. AB - While the selection of amino acid insertions in human immunodeficiency virus (HIV) reverse transcriptase (RT) is a known mechanism of resistance against RT inhibitors, very few reports on the selection of insertions in the protease (PR) coding region have been published. It is still unclear whether these insertions impact protease inhibitor (PI) resistance and/or viral replication capacity. We show that the prevalence of insertions, especially between amino acids 30 to 41 of HIV type 1 (HIV-1) PR, has increased in recent years. We identified amino acid insertions at positions 33 and 35 of the PR of HIV-1-infected patients who had undergone prolonged treatment with PIs, and we characterized the contribution of these insertions to viral resistance. We prepared the corresponding mutated, recombinant PR variants with or without insertions at positions 33 and 35 and characterized them in terms of enzyme kinetics and crystal structures. We also engineered the corresponding recombinant viruses and analyzed the PR susceptibility and replication capacity by recombinant virus assay. Both in vitro methods confirmed that the amino acid insertions at positions 33 and 35 contribute to the viral resistance to most of the tested PIs. The structural analysis revealed local structural rearrangements in the flap region and in the substrate binding pockets. The enlargement of the PR substrate binding site together with impaired flap dynamics could account for the weaker inhibitor binding by the insertion mutants. Amino acid insertions in the vicinity of the binding cleft therefore represent a novel mechanism of HIV resistance development. PMID- 18400859 TI - Partial functional complementation of a pseudorabies virus UL25 deletion mutant by herpes simplex virus type 1 pUL25 indicates overlapping functions of alphaherpesvirus pUL25 proteins. AB - Homologs of the UL25 gene product of herpes simplex virus 1 (HSV-1) are highly conserved among the Herpesviridae. However, their exact function during viral replication is unknown. Current evidence suggests that in the alphaherpesvirus pseudorabies virus (PrV) the capsid-associated pUL25 plays a role in primary envelopment of DNA-containing mature capsids at the inner nuclear membrane. In the absence of pUL25, capsids were found in close association with the inner nuclear membrane, but nuclear egress was not observed (B. G. Klupp, H. Granzow, G. M. Keil, and T. C. Mettenleiter, J. Virol. 80:6235-6246, 2006). In contrast, HSV-1 pUL25 has been assigned a role in stable packaging of viral genomes (N. Stow, J. Virol. 75:10755-10765, 2001). Despite these apparently divergent functions, we wanted to assess whether the high sequence homology translates into functional homology. Therefore, we first analyzed a newly constructed HSV-1 UL25 deletion mutant in our assay system and observed a similar phenotype as in PrV. In the nuclei of infected cells, numerous electron-dense C capsids were detected, whereas primary envelopment of these capsids did not ensue. In agreement with results from PrV, vesicles were observed in the perinuclear space. Since these data indicated functional homology, we analyzed the ability of pUL25 of HSV-1 to complement a PrV UL25 deletion mutant and vice versa. Whereas a HSV-1 pUL25 expressing cell line partially complemented the pUL25 defect in PrV, reciprocal complementation of a HSV-1 UL25 deletion mutant by PrV pUL25 was not observed. Thus, our data demonstrate overlapping, although not identical functions of these two conserved herpesvirus proteins, and point to a conserved functional role in herpes virion formation. PMID- 18400860 TI - During herpes simplex virus type 1 infection of rabbits, the ability to express the latency-associated transcript increases latent-phase transcription of lytic genes. AB - Trigeminal ganglia (TG) from rabbits latently infected with either wild-type herpes simplex virus type 1 (HSV-1) or the latency-associated transcript (LAT) promoter deletion mutant 17DeltaPst were assessed for their viral chromatin profile and transcript abundance. The wild-type 17syn+ genomes were more enriched in the transcriptionally permissive mark dimethyl H3 K4 than were the 17DeltaPst genomes at the 5' exon and ICP0 and ICP27 promoters. Reverse transcription-PCR analysis revealed significantly more ICP4, tk, and glycoprotein C lytic transcripts in 17syn+ than in 17DeltaPst. These results suggest that, for efficient reactivation from latency in rabbits, the LAT is important for increased transcription of lytic genes during latency. PMID- 18400861 TI - Scaffolding proteins altered in the ability to perform a conformational switch confer dominant lethal assembly defects. AB - In the phiX174 procapsid crystal structure, 240 external scaffolding protein D subunits form 60 pairs of asymmetric dimers, D(1)D(2) and D(3)D(4), in a non quasi-equivalent structure. To achieve this arrangement, alpha-helix 3 assumes two different conformations: (i) kinked 30 degrees at glycine residue 61 in subunits D(1) and D(3) and (ii) straight in subunits D(2) and D(4). Substitutions for G61 may inhibit viral assembly by preventing the protein from achieving its fully kinked conformation while still allowing it to interact with other scaffolding and structural proteins. Mutations designed to inhibit conformational switching in alpha-helix 3 were introduced into a cloned gene, and expression was demonstrated to inhibit wild-type morphogenesis. The severity of inhibition appears to be related to the size of the substituted amino acid. For infections in which only the mutant protein is present, morphogenesis does not proceed past the first step that requires the wild-type external scaffolding protein. Thus, mutant subunits alone appear to have little or no morphogenetic function. In contrast, assembly in the presence of wild-type and mutant subunits is blocked prematurely, before D protein is required in a wild-type infection, or channeled into an off-pathway reaction. These data suggest that the wild-type protein transports the inhibitory protein to the pathway. Viruses resistant to the lethal dominant proteins were isolated, and mutations were mapped to the coat and internal scaffolding proteins. The affected amino acids cluster in the atomic structure and may act to exclude mutant subunits from occupying particular positions atop pentamers of the viral coat protein. PMID- 18400862 TI - Small intestine CD4+ T cells are profoundly depleted during acute simian-human immunodeficiency virus infection, regardless of viral pathogenicity. AB - To analyze the relationship between acute virus-induced injury and the subsequent disease phenotype, we compared the virus replication and CD4(+) T-cell profiles for monkeys infected with isogenic highly pathogenic (KS661) and moderately pathogenic (#64) simian-human immunodeficiency viruses (SHIVs). Intrarectal infusion of SHIV-KS661 resulted in rapid, systemic, and massive virus replication, while SHIV-#64 replicated more slowly and reached lower titers. Whereas KS661 systemically depleted CD4(+) T cells, #64 caused significant CD4(+) T-cell depletion only in the small intestine. We conclude that SHIV, regardless of pathogenicity, can cause injury to the small intestine and leads to CD4(+) T cell depletion in infected animals during acute infection. PMID- 18400863 TI - Enhanced pathogenesis of an attenuated herpes simplex virus for mice lacking Stat1. AB - Mice lacking the Stat1 interferon signaling gene were infected with herpes simplex virus type 1 (HSV-1) or an attenuated recombinant lacking virion host shutoff (Delta vhs). Delta vhs virus-infected Stat1(-/-) mice showed levels of replication equivalent to that of the wild-type virus-infected control mice but reduced relative to wild-type virus-infected Stat1(-/-) mice. Stat1 deficiency relieves the immunomodulatory deficiency of Delta vhs virus, but not its inherent growth defect. Also Vhs is dispensable for reactivation. PMID- 18400864 TI - Systematic study of the functions for the residues around the nucleotide pocket in simian virus 40 AAA+ hexameric helicase. AB - The high-resolution structural data for simian virus 40 large-T-antigen helicase revealed a set of nine residues bound to ATP/ADP directly or indirectly. The functional role of each of these residues in ATP hydrolysis and also the helicase function of this AAA+ (ATPases associated with various cellular activities) molecular motor are unclear. Here, we report our mutational analysis of each of these residues to examine their functionality in oligomerization, DNA binding, ATP hydrolysis, and double-stranded DNA (dsDNA) unwinding. All mutants were capable of oligomerization in the presence of ATP and could bind single-stranded DNA and dsDNA. ATP hydrolysis was substantially reduced for proteins with mutations of residues making direct contact with the gamma-phosphate of ATP or the apical water molecule. A potentially noncanonical "arginine finger" residue, K418, is critical for ATP hydrolysis and helicase function, suggesting a new type of arginine finger role by a lysine in the stabilization of the transition state during ATP hydrolysis. Interestingly, our mutational data suggest that the positive- and negative-charge interactions in the uniquely observed residue pairs, R498/D499 and R540/D502, in large-T-antigen helicase are critically involved in the transfer of energy of ATP binding/hydrolysis to DNA unwinding. PMID- 18400865 TI - Site 1 protease is required for proteolytic processing of the glycoproteins of the South American hemorrhagic fever viruses Junin, Machupo, and Guanarito. AB - The cellular proprotein convertase site 1 protease (S1P) has been implicated in the proteolytic processing of the glycoproteins (GPs) of Old World arenaviruses. Here we report that S1P is also involved in the processing of the GPs of the genetically more-distant South American hemorrhagic fever viruses Guanarito, Machupo, and Junin. Efficient cleavage of Guanarito virus GP, whose protease recognition sites deviate from the reported S1P consensus sequence, indicates a broader specificity of S1P than anticipated. Lack of GP processing of Junin virus dramatically reduced production of infectious virus and prevented cell-to-cell propagation. Infection of S1P-deficient cells resulted in viral persistence over several weeks without the emergence of escape variants able to use other cellular proteases for GP processing. PMID- 18400866 TI - In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. AB - Adeno-associated virus (AAV) serotypes differ broadly in transduction efficacies and tissue tropisms and thus hold enormous potential as vectors for human gene therapy. In reality, however, their use in patients is restricted by prevalent anti-AAV immunity or by their inadequate performance in specific targets, exemplified by the AAV type 2 (AAV-2) prototype in the liver. Here, we attempted to merge desirable qualities of multiple natural AAV isolates by an adapted DNA family shuffling technology to create a complex library of hybrid capsids from eight different wild-type viruses. Selection on primary or transformed human hepatocytes yielded pools of hybrids from five of the starting serotypes: 2, 4, 5, 8, and 9. More stringent selection with pooled human antisera (intravenous immunoglobulin [IVIG]) then led to the selection of a single type 2/type 8/type 9 chimera, AAV-DJ, distinguished from its closest natural relative (AAV-2) by 60 capsid amino acids. Recombinant AAV-DJ vectors outperformed eight standard AAV serotypes in culture and greatly surpassed AAV-2 in livers of naive and IVIG immunized mice. A heparin binding domain in AAV-DJ was found to limit biodistribution to the liver (and a few other tissues) and to affect vector dose response and antibody neutralization. Moreover, we report the first successful in vivo biopanning of AAV capsids by using a new AAV-DJ-derived viral peptide display library. Two peptides enriched after serial passaging in mouse lungs mediated the retargeting of AAV-DJ vectors to distinct alveolar cells. Our study validates DNA family shuffling and viral peptide display as two powerful and compatible approaches to the molecular evolution of novel AAV vectors for human gene therapy applications. PMID- 18400867 TI - Cleavage of group 1 coronavirus spike proteins: how furin cleavage is traded off against heparan sulfate binding upon cell culture adaptation. AB - A longstanding enigmatic feature of the group 1 coronaviruses is the uncleaved phenotype of their spike protein, an exceptional property among class I fusion proteins. Here, however, we show that some group 1 coronavirus spike proteins carry a furin enzyme recognition motif and can actually be cleaved, as demonstrated for a feline coronavirus. Interestingly, this feature can be lost during cell culture adaptation by a single mutation in the cleavage motif; this, however, preserves a heparan sulfate binding motif and renders infection by the virus heparan sulfate dependent. We identified a similar cell culture adaptation for the human coronavirus OC43. PMID- 18400868 TI - AT1 blockade during lactation as a model of chronic nephropathy: mechanisms of renal injury. AB - Suppression of the renin-angiotensin system during lactation causes irreversible renal structural changes. In this study we investigated 1) the time course and the mechanisms underlying the chronic kidney disease caused by administration of the AT(1) receptor blocker losartan during lactation, and 2) whether this untoward effect can be used to engender a new model of chronic kidney disease. Male Munich-Wistar pups were divided into two groups: C, whose mothers were untreated, and L(Lact), whose mothers received oral losartan (250 mg.kg(-1).day( 1)) during the first 20 days after delivery. At 3 mo of life, both nephron number and the glomerular filtration rate were reduced in L(Lact) rats, whereas glomerular pressure was elevated. Unselective proteinuria and decreased expression of the zonula occludens-1 protein were also observed, along with modest glomerulosclerosis, significant interstitial expansion and inflammation, and wide glomerular volume variation, with a stable subpopulation of exceedingly small glomeruli. In addition, the urine osmolality was persistently lower in L(Lact) rats. At 10 mo of age, L(Lact) rats exhibited systemic hypertension, heavy albuminuria, substantial glomerulosclerosis, severe renal interstitial expansion and inflammation, and creatinine retention. Conclusions are that 1) oral losartan during lactation can be used as a simple and easily reproducible model of chronic kidney disease in adult life, associated with low mortality and no arterial hypertension until advanced stages; and 2) the mechanisms involved in the progression of renal injury in this model include glomerular hypertension, glomerular hypertrophy, podocyte injury, and interstitial inflammation. PMID- 18400869 TI - Cytoplasmic initiation of cisplatin cytotoxicity. AB - The mechanism of action of cisplatin as a chemotherapeutic agent has been attributed to DNA binding, while its mechanism of action as a nephrotoxin is unresolved. Only approximately 1% of intracellular cisplatin interacts with DNA, primarily forming intrastrand cross-linked adducts, and many studies have implicated both nuclear and cytoplasmic causes of cisplatin-induced death in cultured cells. We have demonstrated that cisplatin cytotoxicity depends on cdk2 activity, which is at least partly through the cdk2-E2F1 pathway. The mechanism of the dependency on cdk2, and whether cdk2 activation of E2F1 represents the only cell death pathway involved, is still unclear. Our previous work showed that deletion of the nuclear localization signal from p21 WAF1/CIP1, a cdk2 inhibitor, did not alter its protective action against cisplatin cytotoxicity. Active cdk2 cyclin complexes are localized in both the nucleus and cytoplasm, and it was reported that cdk2 translocated to the cytoplasm after an apoptotic stimulus. Herein, we show that cisplatin caused cell death in enucleated mouse kidney proximal tubule cells (TKPTS), which was prevented by cdk2 inhibition. Also, we localized cytoplasmic cdk2 to both the endoplasmic reticulum (ER) and Golgi compartments, and ER stress was blocked by specific cdk2 inhibition. We conclude that cisplatin can induce nuclear independent apoptosis, cisplatin cytotoxicity can be initiated by cytoplasmic events, and cytoplasmic cdk2 plays an important role in apoptosis signaling. PMID- 18400870 TI - Exploring mechanisms involved in renal tubular sensing of mechanical stretch following ureteric obstruction. AB - Tubular mechanical stretch is the key primary insult in obstructive nephropathy. This review addresses how the renal tubular epithelium senses and responds to mechanical stretch. Using data from renal and nonrenal systems, we describe how sensing of stretch initially occurs via the activation of ion channels and subsequent increases in intracellular calcium levels. Calcium influxes activate a number of adaptive and proinjury responses. Key among these are 1) the activation of Rho, consequent cytoskeletal rearrangements, and downstream increases in focal adhesion assembly; and 2) phospholipase activation and resultant mitogen activated protein kinase activation. These early signaling events culminate in adaptive cellular coupling to the extracellular matrix, a process termed the cell strengthening response. Direct links can be made between increased expression of genes involved in the development of obstructive nephropathy and initial sensing of mechanical stretch. The review illustrates the repercussions of mechanical stretch as a renal stress stimulus, specific to ureteric obstruction, and provides an insight into how tubular responses to mechanical stretch are ultimately implicated in the development of obstructive nephropathy. PMID- 18400871 TI - Renal hemodynamic effect of cyclooxygenase 2 inhibition in young men and women with uncomplicated type 1 diabetes mellitus. AB - In experimental studies, cyclooxygenase 2 (COX2)-derived vasodilatory prostaglandins play a more prominent role in arterial vasoregulation in females. The gender-dependent effect of COX2 modulation in humans with type 1 diabetes mellitus (DM) is unknown. Accordingly, we examined the renal hemodynamic role of prostaglandins by assessing the response to COX2 inhibition in young men and women with type 1 DM. We also used a graded ANG II infusion to determine whether gender-based differences were mediated by effects of COX2 inhibition on the renin angiotensin system (RAS). We hypothesized that COX2 inhibition would be associated with preferential vasoconstriction in women and would augment their response to ANG II. Baseline renal function and the response to an ANG II infusion were assessed during clamped euglycemia, and again after COX2 inhibition (200 mg celecoxib daily for 14 days) in 12 men and 9 women after 1 wk on a controlled protein and sodium diet. COX2 inhibition was associated with increases in filtration fraction (P = 0.045) and renal vascular resistance and a decline in renal blood flow (P = 0.04) in women compared with men. Before COX2 inhibition, women exhibited a decline in glomerular filtration rate in response to ANG II. COX2 inhibition abolished this effect, whereas the response was not altered in men. In summary, COX2 inhibition was associated with hemodynamic effects that differed based on gender. The ANG II response suggests that with uncomplicated type 1 DM, prostaglandins may contribute to RAS-mediated gender differences. Our results are consistent with experimental data suggesting augmented female prostanoid dependence. PMID- 18400872 TI - Effect of renal denervation on prenatal programming of hypertension and renal tubular transporter abundance. AB - Prenatal glucocorticoids are often administered to pregnant women to accelerate pulmonary maturation. We have demonstrated that administration of dexamethasone during specific periods of pregnancy in the rat causes hypertension in the offspring when they are studied as adults. The purpose of the present study was to determine whether the hypertension due to prenatal dexamethasone was mediated by renal nerves. We administered dexamethasone to rats daily for 4 days between days 15 and 18 of gestation. Rats underwent bilateral renal denervation or sham operation at 6 wk of age, and blood pressure was measured at 8 wk of age. Prenatal dexamethasone in the sham operation group resulted in an increase in blood pressure compared with vehicle-treated sham controls (134 +/- 3 vs. 145 +/- 3 mmHg, P < 0.05). Renal denervation did not affect blood pressure significantly in the prenatal vehicle-treated control group but resulted in normalization in blood pressure in the prenatal dexamethasone group and (130 +/- 3 and 128 +/- 5 mmHg, respectively). Prenatal dexamethasone increased type 3 Na+/H+ exchanger (NHE3), Na+K+-2Cl(-) cotransporter (NKCC2), and Na+-Cl(-) cotransporter (NCC), but not alpha-, beta-, and gamma-epithelial Na+ channel (ENaC) protein abundance compared with controls. The increase in NHE3, NKCC2, and NCC protein abundance by prenatal dexamethasone was not seen in 8-wk-old rats 2 wk after renal denervation. Renal denervation did not affect NHE3, NKCC2, and NCC protein abundance in prenatal vehicle-treated animals. This study is consistent with renal nerves playing a role in mediating the hypertension by prenatal programming by dexamethasone. PMID- 18400874 TI - A highly sensitive technique to measure myosin regulatory light chain phosphorylation: the first quantification in renal arterioles. AB - Phosphorylation of the 20-kDa myosin regulatory light chains (LC(20)) plays a key role in the regulation of smooth muscle contraction. The level of LC(20) phosphorylation is governed by the relative activities of myosin light chain kinase and phosphatase pathways. The regulation of these two pathways differs in different smooth muscle types and in the actions of different vasoactive stimuli. Little is known concerning the regulation of LC(20) phosphorylation in the renal microcirculation. The available pharmacological probes are often nonspecific, and current techniques to directly measure LC(20) phosphorylation are not sensitive enough for quantification in small arterioles. We describe here a novel approach to address this important issue. Using SDS-PAGE with polyacrylamide-bound Mn(2+) phosphate-binding tag and enhanced Western blot analysis, we were able to detect LC(20) phosphorylation using as little as 5 pg (250 amol) of isolated LC(20). Phosphorylated and unphosphorylated LC(20) were detected in single isolated afferent arterioles, and LC(20) phosphorylation levels could be accurately quantified in pooled samples of three arterioles (<300 cells). The phosphorylation level of LC(20) in the afferent arteriole was 6.8 +/- 1.7% under basal conditions and increased to 34.7 +/- 5.1% and 44.6 +/- 6.6% in response to 30 mM KCl and 10(-8) M angiotensin II, respectively. The application of this technique will enable investigations of the different determinants of LC(20) phosphorylation in afferent and efferent arterioles and provide insights into the signaling pathways that regulate LC(20) phosphorylation in the renal microvasculature under physiological and pathophysiological conditions. PMID- 18400873 TI - The natriuretic and diuretic response to dopamine is maintained during rat pregnancy. AB - During pregnancy, there is a marked plasma volume expansion due to renal sodium retention. Pregnant rats exhibit a blunted response to natriuretic stimuli that signal via cGMP, and expression and activity of the cGMP phosphodiesterase PDE-5 are upregulated in the inner medullary collecting duct during pregnancy. Here, we tested the hypothesis that the natriuretic response to a cAMP agonist, dopamine, is maintained during pregnancy. Anesthetized pregnant (day 16) and age-matched virgin Sprague-Dawley rats were used to determine whether dopamine-cAMP-mediated natriuresis remains intact in pregnant rats. Blood pressure, renal clearances of inulin and p-aminohippuric acid, and excretion of sodium were measured during baseline and dopamine infusion periods. Pregnant rats had a lower blood pressure and hematocrit at baseline than their age-matched virgin counterparts. Dopamine infusion decreased blood pressure and increased glomerular filtration rate and renal plasma flow in virgin but not pregnant rats. Dopamine infusion also increased urine volume, sodium excretion, and the fractional excretion of sodium to a similar extent in virgin and pregnant rats. These results indicate that a cAMP-mediated natriuresis and diuresis (stimulated by dopamine) persists in pregnant rats. PMID- 18400875 TI - Effective prevention of sports injuries: a model integrating efficacy, efficiency, compliance and risk-taking behaviour. AB - In 1992 van Mechelen et al published a "sequence of prevention model" based on a four-step process. This model has been widely used to implement preventive measures in response to sports injuries. However, the execution of this model has been shown to lack validity in determining the success of a preventive measure. A modified model has therefore been proposed which incorporates additional steps that enable the inclusion of external factors with a significant effect on the outcome of a prevention intervention. This expansion of van Mechelen's model leads to a more global model in which the compliance level and risk-taking behaviour of the individual and the assessment of efficiency of the stakeholders have a key influence on the preventive measure. This model gives a better insight into the different processes in injury prevention that can be used by clinicians, coaches and managers to decide whether to implement a preventive measure programme. PMID- 18400877 TI - Mobilisation of mesenchymal cells in cardiac patients: is intense exercise necessary? AB - Circulating mesenchymal cells (cMCs) have a potential for regenerating damaged tissue, e.g., ischaemic myocardium. In patients (age range: 53-76 years) with stable coronary artery disease cMCs were determined before and after dynamic exercise of moderate (< respiratory compensation threshold (RCT)) (n = 9 patients) or high intensity (>RCT) (n = 11). Only high-intensity exercise (i.e., provoking signs of myocardial ischaemia in 3 patients and ventricular extrasystoles in another) induced a significant increase in cMCs (p = 0.009). These results support the hypothesis that intense exercise (near or at the point of myocardial ischaemia) is a potent stimulus for MC mobilisation. PMID- 18400876 TI - Drinking policies and exercise-associated hyponatraemia: is anyone still promoting overdrinking? AB - OBJECTIVES: The purpose of this review is to describe the evolution of hydration research and advice on drinking during exercise from published scientific papers, books and non-scientific material (advertisements and magazine contents) and detail how erroneous advice is likely propagated throughout the global sports medicine community. DESIGN: Hydration advice from sports-linked entities, the scientific community, exercise physiology textbooks and non-scientific sources was analysed historically and compared with the most recent scientific evidence. CONCLUSIONS: Drinking policies during exercise have changed substantially throughout history. Since the mid-1990s, however, there has been an increase in the promotion of overdrinking by athletes. While the scientific community is slowly moving away from "blanket" hydration advice in which one form of advice fits all and towards more modest, individualised, hydration guidelines in which thirst is recognised as the best physiological indicator of each subject's fluid needs during exercise, marketing departments of the global sports drink industry continue to promote overdrinking. PMID- 18400878 TI - The use of prolotherapy in the sacroiliac joint. AB - OBJECTIVE: In this study the effectiveness of prolotherapy in the treatment of deficient load transfer of the sacroiliac joint (SIJ) was determined. DESIGN: A prospective descriptive study. SETTING: Authors' private practice. PARTICIPANTS: 25 patients who consented to treatment and attended for at least one follow-up visit and assessment. STUDY PERIOD: From April 2004 to July 2007. INTERVENTION: Three injections of hypertonic dextrose solution into the dorsal interosseous ligament of the affected SIJ, under CT control, 6 weeks apart. MAIN OUTCOME MEASURES: Quebec Back Pain Disability Scale, Roland-Morris 24, Roland-Morris 24 Multiform questionnaires and clinical examination by two authors independently. RESULTS: All patients included in this study attended at least one follow-up visit at 3, 12 or 24 months.. The number of patients at follow-up decreased at 12 and 24 months. Functional questionnaires demonstrated significant improvements for those followed-up at 3, 12 and 24 months (p<0.05). Clinical scores showed significant improvement from commencement to 3, 12 and 24 months (p<0.001). CONCLUSIONS: This descriptive study of prolotherapy in private practice has shown positive clinical outcomes for the 76% of patients who attended the 3-month follow-up visit (76% at 12 months and 32% at 24 months). Similar results were found in the questionnaires (Quebec Back Pain Disability Scale, Roland-Morris 24 and Roland-Morris 24 Multiform questionnaires) at 3, 12 and 24 months. PMID- 18400879 TI - Coeliac disease and the risk of infections. PMID- 18400880 TI - High school coaches' assessments, intentions to use, and use of a concussion prevention toolkit: Centers for Disease Control and Prevention's heads up: concussion in high school sports. AB - This study evaluated school coaches' perceptions, assessments, and use of a toolkit to prevent and manage concussions among school athletes. A computer assisted telephone survey was conducted with a stratified, random sample of high school coaches (n = 497; response rate = 39.3%; cooperation rate = 81.5%) from five states. Most reported that they had used or planned to use kit materials. Most (81%) in schools with a written plan for preventing and managing concussions indicated that the toolkit could be used to improve it and 96% of coaches in schools without a plan indicated that the kit could be used to develop one. Most assessed the kit as visually appealing, easy to use, and containing appropriate content. There were no significant differences among coaches with differing professional experience or for sports with different injury rates. Among those with other concussion-prevention materials, most indicated greater satisfaction with the toolkit. PMID- 18400881 TI - Transcription factor Lim1 specifies horizontal cell laminar position in the retina. PMID- 18400882 TI - Short-term memory and the human hippocampus. PMID- 18400883 TI - Insolubility of disrupted-in-schizophrenia 1 disrupts oligomer-dependent interactions with nuclear distribution element 1 and is associated with sporadic mental disease. AB - Disrupted-in-schizophrenia 1 (DISC1) and other genes have been identified recently as potential molecular players in chronic psychiatric diseases such as affective disorders and schizophrenia. A molecular mechanism of how these genes may be linked to the majority of sporadic cases of these diseases remains unclear. The chronic nature and irreversibility of clinical symptoms in a subgroup of these diseases prompted us to investigate whether proteins corresponding to candidate genes displayed subtle features of protein aggregation. Here, we show that in postmortem brain samples of a distinct group of patients with phenotypes of affective disorders or schizophrenia, but not healthy controls, significant fractions of DISC1 could be identified as cold Sarkosyl-insoluble protein aggregates. A loss-of-function phenotype could be demonstrated for insoluble DISC1 through abolished binding to a key DISC1 ligand, nuclear distribution element 1 (NDEL1): in human neuroblastoma cells, DISC1 formed expression-dependent, detergent-resistant aggregates that failed to interact with endogenous NDEL1. Recombinant (r) NDEL1 expressed in Escherichia coli selectively bound an octamer of an rDISC1 fragment but not dimers or high molecular weight multimers, suggesting an oligomerization optimum for molecular interactions of DISC1 with NDEL1. For DISC1-related sporadic psychiatric disease, we propose a mechanism whereby impaired cellular control over self-association of DISC1 leads to excessive multimerization and subsequent formation of detergent resistant aggregates, culminating in loss of ligand binding, here exemplified by NDEL1. We conclude that the absence of oligomer-dependent ligand interactions of DISC1 can be associated with sporadic mental disease of mixed phenotypes. PMID- 18400884 TI - Reverse signaling via a glycosyl-phosphatidylinositol-linked ephrin prevents midline crossing by migratory neurons during embryonic development in Manduca. AB - We have investigated whether reverse signaling via a glycosyl phosphatidylinositol (GPI)-linked ephrin controls the behavior of migratory neurons in vivo. During the formation of the enteric nervous system (ENS) in the moth Manduca, approximately 300 neurons [enteric plexus (EP) cells] migrate onto the midgut via bilaterally paired muscle bands but avoid adjacent midline regions. As they migrate, the EP cells express a single ephrin ligand (MsEphrin; a GPI-linked ligand), whereas the midline cells express the corresponding Eph receptor (MsEph). Blocking endogenous MsEphrin-MsEph receptor interactions in cultured embryos resulted in aberrant midline crossing by the neurons and their processes. In contrast, activating endogenous MsEphrin on the EP cells with dimeric MsEph-Fc constructs inhibited their migration and outgrowth, supporting a role for MsEphrin-dependent reverse signaling in this system. In short-term cultures, blocking endogenous MsEph receptors allowed filopodia from the growth cones of the neurons to invade the midline, whereas activating neuronal MsEphrin led to filopodial retraction. MsEphrin-dependent signaling may therefore guide the migratory enteric neurons by restricting the orientation of their leading processes. Knocking down MsEphrin expression in the EP cells with morpholino antisense oligonucleotides also induced aberrant midline crossing, consistent with the effects of blocking endogenous MsEphrin-MsEph interactions. Unexpectedly, this treatment enhanced the overall extent of migration, indicating that MsEphrin-dependent signaling may also modulate the general motility of the EP cells. These results demonstrate that MsEphrin-MsEph receptor interactions normally prevent midline crossing by migratory neurons within the developing ENS, an effect that is most likely mediated by reverse signaling through this GPI linked ephrin ligand. PMID- 18400886 TI - Alterations in Ca2+-buffering in prion-null mice: association with reduced afterhyperpolarizations in CA1 hippocampal neurons. AB - Prion protein (PrP) is a normal component of neurons, which confers susceptibility to prion diseases. Despite its evolutionary conservation, its normal function remains controversial. PrP-deficient (Prnp(0/0)) mice have weaker afterhyperpolarizations (AHPs) in cerebellar and hippocampal neurons. Here we show that the AHP impairment in hippocampal CA1 pyramidal cells is selective for the slow AHP, and is not caused by an impairment of either voltage-gated Ca(2+) channels or Ca(2+)-activated K(+) channels. Instead, Prnp(0/0) neurons have twofold to threefold stronger Ca(2+) buffering and double the Ca(2+) extrusion rate. In Prnp(0/0) neurons thapsigargin abolished the stronger Ca(2+) buffering and extrusion, and thapsigargin or cyclopiazonic acid abolished the weakening of the slow AHPs. These data implicate sarcoplasmic/endoplasmic reticulum calcium ATPase in the enhanced Ca(2+) buffering, and extrusion into the endoplasmic reticulum, which contains substantial amounts of PrP in wild-type mice. Altered Ca(2+) homeostasis can explain several phenotypes identified in Prnp(0/0) mice. PMID- 18400885 TI - Differential mechanisms of CRF1 and CRF2 receptor functions in the amygdala in pain-related synaptic facilitation and behavior. AB - A major site of extrahypothalamic expression of corticotropin-releasing factor (CRF) and its G-protein-coupled CRF1 and CRF2 receptors is the amygdala, a key player in emotions and affective disorders. Pain-related plasticity in the laterocapsular division of the central nucleus of the amygdala (CeLC) generates emotional-affective responses and anxiety-like behavior. CRF1 receptor antagonists have anxiolytic effects. Although both CRF1 and CRF2 receptors couple positively to adenylyl cyclase, they can have opposite effects, but the underlying mechanism is unknown. This study addressed CRF1 and CRF2 receptor functions and mechanisms in the amygdala in a model of arthritic pain. Using whole-cell patch-clamp recordings of CeLC neurons, we found that a selective CRF1 receptor antagonist (NBI27914 [5-chloro-4-(N-(cyclopropyl)methyl-N-propylamino)-2 methyl-6-(2,4,6-trichlorophenyl)]) amino-pyridine inhibited synaptic facilitation in brain slices from arthritic rats through a postsynaptic mechanism. Inhibition of the NMDA receptor-mediated synaptic component was occluded by a protein kinase A (PKA) inhibitor, consistent with our previous demonstration of PKA-dependent increased NMDA receptor function in arthritis pain-related plasticity. NBI27914 also decreased neuronal excitability through inhibition of highly tetraethylammonium (TEA)-sensitive ion channels that contribute to action potential repolarization and firing rate. In contrast, a CRF2 receptor antagonist (astressin-2B [cyclo(31-34) [d-Phe11,His12,C alphaMeLeu13,39, Nle17, Glu31, Lys34] Ac-Sauvagine(8-40)]) facilitated synaptic transmission through presynaptic inhibition of GABAergic transmission (disinhibition). NBI27914 inhibited arthritis pain-related behaviors (audible and ultrasonic vocalizations and hindlimb withdrawal reflexes). Astressin-2B had no significant behavioral effect. The data suggest that endogenous CRF1 receptor activation in the amygdala contributes to pain-related synaptic facilitation, increased excitability, and pain behavior through a postsynaptic mechanism involving activation of PKA and highly TEA-sensitive K(+)-currents. Presynaptic CRF2 receptor-mediated inhibition does not reach behavioral significance. PMID- 18400887 TI - No evidence for chronic demyelination in spared axons after spinal cord injury in a mouse. AB - The pattern of remyelination after traumatic spinal cord injury remains elusive, with animal and human studies reporting partial to complete demyelination followed by incomplete remyelination. In the present study, we found that spared rubrospinal tract (RST) axons of passage traced with actively transported dextrans and examined caudally to the lesion 12 weeks after mouse spinal cord contusion injury were fully remyelinated. Spared axons exhibited a marginally reduced myelin thickness and significantly shorter internodes. CASPR (contactin associated protein) and K(v)1.2 channels were used to identify internodes and paranodal protein distribution properties were used as an index of myelin integrity. This is the first time the CNS myelin internode length was measured in a mouse. To better understand the significance of shortened internodes and thinner myelin in spared axons, we modeled conduction properties using McIntyre's et al. model of myelinated axons. Mathematical modeling predicted a 21% decrease in the conduction velocity of remyelinated RST axons attributable to shortened internodes. To determine whether demyelination could be present on axons exhibiting a pathological transport system, we used the retroviral reporter system. Virally delivered green fluorescent protein unveiled a small population of dystrophic RST axons that persist chronically with evident demyelination or abnormal remyelination. Collectively, these data show that lasting demyelination in spared axons is rare and that remyelination of axons of passage occurs in the chronically injured mouse spinal cord. PMID- 18400889 TI - Impaired adult neurogenesis associated with short-term memory defects in NF kappaB p50-deficient mice. AB - Neurogenesis proceeds throughout adulthood in the brain of most mammalian species, but the molecular mechanisms underlying the regulation of stem/progenitor cell proliferation, survival, maturation, and differentiation have not been completely unraveled. We have studied hippocampal neurogenesis in NF-kappaB p50-deficient mice. Here we demonstrate that in absence of p50, the net rate of neural precursor proliferation does not change, but some of the steps leading to the final neuron differentiation status are hampered, resulting in approximately 50% reduction in the number of newly born neurons in the adult mutant hippocampus. Additionally, in p50(-/-) mice, we observed a selective defect in short-term spatial memory performance without impairment of hippocampal dependent spatial long-term memory and learning. Our results highlight the role of NF-kappaB p50 in hippocampal neurogenesis and in short-term spatial memory. PMID- 18400890 TI - Soluble adenylyl cyclase is not required for axon guidance to netrin-1. AB - During development, axons are directed to their targets by extracellular guidance cues. The axonal response to the guidance cue netrin-1 is profoundly influenced by the concentration of cAMP within the growth cone. In some cases, cAMP affects the sensitivity of the growth cone to netrin-1, whereas in others it changes the response to netrin-1 from attraction to repulsion. The effects of cAMP on netrin 1 action are well accepted, but the critical issue of whether cAMP production is activated by a netrin-1 induced signaling cascade remains uncertain. A previous report has suggested that axon guidance in response to netrin-1 requires cAMP production mediated by soluble adenyl cyclase (sAC). We have used genetic, molecular and biochemical strategies to assess this issue. Surprisingly, we found only extremely weak expression of sAC in embryonic neurons and determined that, under conditions where netrin-1 directs axonal pathfinding, exposure to netrin-1 does not alter cAMP levels. Furthermore, although netrin-1-deficient mice exhibit major axon guidance defects, we show that pathfinding is normal in sAC-null mice. Therefore, although cAMP can alter the response of axons to netrin-1, we conclude that netrin-1 does not alter cAMP levels in axons attracted by this cue, and that sAC is not required for axon attraction to netrin-1. PMID- 18400888 TI - Spectrotemporal processing differences between auditory cortical fast-spiking and regular-spiking neurons. AB - Excitatory pyramidal neurons and inhibitory interneurons constitute the main elements of cortical circuitry and have distinctive morphologic and electrophysiological properties. Here, we differentiate them by analyzing the time course of their action potentials (APs) and characterizing their receptive field properties in auditory cortex. Pyramidal neurons have longer APs and discharge as regular-spiking units (RSUs), whereas basket and chandelier cells, which are inhibitory interneurons, have shorter APs and are fast-spiking units (FSUs). To compare these neuronal classes, we stimulated cat primary auditory cortex neurons with a dynamic moving ripple stimulus and constructed single-unit spectrotemporal receptive fields (STRFs) and their associated nonlinearities. FSUs had shorter latencies, broader spectral tuning, greater stimulus specificity, and higher temporal precision than RSUs. The STRF structure of FSUs was more separable, suggesting more independence between spectral and temporal processing regimens. The nonlinearities associated with the two cell classes were indicative of higher feature selectivity for FSUs. These global functional differences between RSUs and FSUs suggest fundamental distinctions between putative excitatory and inhibitory interneurons that shape auditory cortical processing. PMID- 18400891 TI - Perturbation of syndapin/PACSIN impairs synaptic vesicle recycling evoked by intense stimulation. AB - Synaptic vesicle recycling has been proposed to depend on proteins which coordinate membrane and cytoskeletal dynamics. Here, we examine the role of the dynamin- and N-WASP (neural Wiskott-Aldrich syndrome protein)-binding protein syndapin/PACSIN at the lamprey reticulospinal synapse. We find that presynaptic microinjection of syndapin antibodies inhibits vesicle recycling evoked by intense (5 Hz or more), but not by light (0.2 Hz) stimulation. This contrasts with the inhibition at light stimulation induced by perturbation of amphiphysin (Shupliakov et al., 1997). Inhibition by syndapin antibodies was associated with massive accumulation of membranous cisternae and invaginations around release sites, but not of coated pits at the plasma membrane. Cisternae contained vesicle membrane, as shown by vesicle-associated membrane protein 2 (VAMP2)/synaptobrevin 2 immunolabeling. Similar effects were observed when syndapin was perturbed before onset of massive endocytosis induced by preceding intense stimulation. Selective perturbation of the Src homology 3 domain interactions of syndapin was sufficient to induce vesicle depletion and accumulation of cisternae. Our data show an involvement of syndapin in synaptic vesicle recycling evoked by intense stimulation. We propose that syndapin is required to stabilize the plasma membrane and/or facilitate bulk endocytosis at high release rates. PMID- 18400892 TI - Prefrontal cortex function in the representation of temporally complex events. AB - The frontal cortex and inferior temporal cortex are strongly functionally interconnected. Previous experiments on prefrontal function in monkeys have shown that a disconnection of prefrontal cortex from inferior temporal cortex impairs a variety of complex visual learning tasks but leaves simple concurrent object reward association learning intact. We investigated the possibility that temporal components of visual learning tasks determine the sensitivity of those tasks to prefrontal-temporal disconnection by adding specific temporal components to the concurrent object-reward association learning task. Monkeys with crossed unilateral lesions of prefrontal cortex and inferior temporal cortex were impaired compared with unoperated controls at associating two-item sequences of visual objects with reward. The impairment was specific to the learning of visual sequences, because disconnection was without effect on object-reward association learning for an equivalent delayed reward. This result was replicated in monkeys with transection of the uncinate fascicle, thus determining the anatomical specificity of the dissociation. Previous behavioral results suggest that monkeys represent the two-item serial compound stimuli in a configural manner, similar to the way monkeys represent simultaneously presented compound stimuli. The representation of simultaneously presented configural stimuli depends on the perirhinal cortex. The present experiments show that the representation of serially presented compound stimuli depends on the interaction of prefrontal cortex and inferior temporal cortex. We suggest that prefrontal-temporal disconnection impairs a wide variety of learning tasks because in those tasks monkeys lay down similar temporally complex representations. PMID- 18400893 TI - Beta-amyloid(1-42) induces neuronal death through the p75 neurotrophin receptor. AB - Alzheimer's disease is characterized by the accumulation of neurotoxic amyloidogenic peptide Abeta, degeneration of the cholinergic innervation to the hippocampus (the septohippocampal pathway), and progressive impairment of cognitive function, particularly memory. Abeta is a ligand for the p75 neurotrophin receptor (p75(NTR)), which is best known for mediating neuronal death and has been consistently linked to the pathology of Alzheimer's disease. Here we examined whether p75(NTR) is required for Abeta-mediated effects. Treatment of wild-type but not p75(NTR)-deficient embryonic mouse hippocampal neurons with human Abeta(1-42) peptide induced significant cell death. Furthermore, injection of Abeta(1-42) into the hippocampus of adult mice resulted in significant degeneration of wild-type but not p75(NTR)-deficient cholinergic basal forebrain neurons, indicating that the latter are resistant to Abeta induced toxicity. We also found that neuronal death correlated with Abeta(1-42) peptide-stimulated accumulation of the death-inducing p75(NTR) C-terminal fragment generated by extracellular metalloprotease cleavage of full-length p75(NTR). Although neuronal death was prevented in the presence of the metalloprotease inhibitor TAPI-2 (tumor necrosis factor-alpha protease inhibitor 2), Abeta(1-42)-induced accumulation of the C-terminal fragment resulted from inhibition of gamma-secretase activity. These results provide a novel mechanism to explain the early and characteristic loss of cholinergic neurons in the septohippocampal pathway that occurs in Alzheimer's disease. PMID- 18400894 TI - Altered histone monoubiquitylation mediated by mutant huntingtin induces transcriptional dysregulation. AB - Although transcriptional dysregulation is a critical pathogenic mechanism in Huntington's disease (HD), it is still not known how mutant huntingtin causes it. Here we show that alteration of histone monoubiquitylation is a key mechanism. Disrupted interaction of huntingtin with Bmi-1, a component of the hPRC1L E3 ubiquitin ligase complex, increases monoubiquityl histone H2A (uH2A) levels in a cell culture model of HD. Genes with expression that is repressed in transgenic R6/2 mouse brain have increased uH2A and decreased uH2B at their promoters, whereas actively transcribed genes show the opposite pattern. Reduction in uH2A reverses transcriptional repression and inhibits methylation of histone H3 at lysine 9 in cell culture. In contrast, reduction in uH2B induces transcriptional repression and inhibits methylation of histone H3 at lysine 4. This is the first report to demonstrate hPRC1L as a huntingtin-interacting histone modifying complex and a crucial role for histone monoubiquitylation in mammalian brain gene expression, which broadens our understanding of histone code. These findings also provide a rationale for targeting histone monoubiquitylation for therapy in HD. PMID- 18400895 TI - Right-hemisphere auditory cortex is dominant for coding syllable patterns in speech. AB - Cortical analysis of speech has long been considered the domain of left hemisphere auditory areas. A recent hypothesis poses that cortical processing of acoustic signals, including speech, is mediated bilaterally based on the component rates inherent to the speech signal. In support of this hypothesis, previous studies have shown that slow temporal features (3-5 Hz) in nonspeech acoustic signals lateralize to right-hemisphere auditory areas, whereas rapid temporal features (20-50 Hz) lateralize to the left hemisphere. These results were obtained using nonspeech stimuli, and it is not known whether right hemisphere auditory cortex is dominant for coding the slow temporal features in speech known as the speech envelope. Here we show strong right-hemisphere dominance for coding the speech envelope, which represents syllable patterns and is critical for normal speech perception. Right-hemisphere auditory cortex was 100% more accurate in following contours of the speech envelope and had a 33% larger response magnitude while following the envelope compared with the left hemisphere. Asymmetries were evident regardless of the ear of stimulation despite dominance of contralateral connections in ascending auditory pathways. Results provide evidence that the right hemisphere plays a specific and important role in speech processing and support the hypothesis that acoustic processing of speech involves the decomposition of the signal into constituent temporal features by rate-specialized neurons in right- and left-hemisphere auditory cortex. PMID- 18400896 TI - The distinct temporal origins of olfactory bulb interneuron subtypes. AB - Olfactory bulb (OB) interneurons are a heterogeneous population produced beginning in embryogenesis and continuing through adulthood. Understanding how this diversity arises will provide insight into how OB microcircuitry is established as well as adult neurogenesis. Particular spatial domains have been shown to contribute specific interneuron subtypes. However, the temporal profile by which OB interneuron subtypes are produced is unknown. Using inducible genetic fate mapping of Dlx1/2 precursors, we analyzed the production of seven OB interneuron subtypes and found that the generation of each subpopulation has a unique temporal signature. Within the glomerular layer, the production of tyrosine hydroxylase-positive interneurons is maximal during early embryogenesis and decreases thereafter. In contrast, the generation of calbindin interneurons is maximal during late embryogenesis and declines postnatally, whereas calretinin (CR) cell production is low during embryogenesis and increases postnatally. Parvalbumin interneurons within the external plexiform layer are produced only perinatally, whereas the generation of 5T4-positive granule cells in the mitral cell layer does not change significantly over time. CR-positive granule cells are not produced at early embryonic time points, but constitute a large percentage of the granule cells born after birth. Blanes cells in contrast are produced in greatest number during embryogenesis. Together we provide the first comprehensive analysis of the temporal generation of OB interneuron subtypes and demonstrate that the timing by which these populations are produced is tightly orchestrated. PMID- 18400897 TI - Prominent role of the spinal central pattern generator in the recovery of locomotion after partial spinal cord injuries. AB - The re-expression of hindlimb locomotion after complete spinal cord injuries (SCIs) is caused by the presence of a spinal central pattern generator (CPG) for locomotion. After partial SCI, however, the role of this spinal CPG in the recovery of hindlimb locomotion in the cat remains mostly unknown. In the present work, we devised a dual-lesion paradigm to determine its possible contribution after partial SCI. After a partial section of the left thoracic segment T10 or T11, cats gradually recovered voluntary quadrupedal locomotion. Then, a complete transection was performed two to three segments more caudally (T13-L1) several weeks after the first partial lesion. Cats that received intensive treadmill training after the partial lesion expressed bilateral hindlimb locomotion within hours of the complete lesion. Untrained cats however showed asymmetrical hindlimb locomotion with the limb on the side of the partial lesion walking well before the other hindlimb. Thus, the complete spinalization revealed that the spinal CPG underwent plastic changes after the partial lesions, which were shaped by locomotor training. Over time, with further treadmill training, the asymmetry disappeared and a bilateral locomotion was reinstated. Therefore, although remnant intact descending pathways must contribute to voluntary goal-oriented locomotion after partial SCI, the recovery and re-expression of the hindlimb locomotor pattern mostly results from intrinsic changes below the lesion in the CPG and afferent inputs. PMID- 18400898 TI - Maps of visual space in human occipital cortex are retinotopic, not spatiotopic. AB - We experience the visual world as phenomenally invariant to eye position, but almost all cortical maps of visual space in monkeys use a retinotopic reference frame, that is, the cortical representation of a point in the visual world is different across eye positions. It was recently reported that human cortical area MT (unlike monkey MT) represents stimuli in a reference frame linked to the position of stimuli in space, a "spatiotopic" reference frame. We used visuotopic mapping with blood oxygen level-dependent functional magnetic resonance imaging signals to define 12 human visual cortical areas, and then determined whether the reference frame in each area was spatiotopic or retinotopic. We found that all 12 areas, including MT, represented stimuli in a retinotopic reference frame. Although there were patches of cortex in and around these visual areas that were ostensibly spatiotopic, none of these patches exhibited reliable stimulus-evoked responses. We conclude that the early, visuotopically organized visual cortical areas in the human brain (like their counterparts in the monkey brain) represent stimuli in a retinotopic reference frame. PMID- 18400899 TI - Developmental plasticity in the human auditory brainstem. AB - Development of the human auditory brainstem is thought to be primarily complete by the age of approximately 2 years, such that subsequent sensory plasticity is confined primarily to the cortex. However, recent findings have revealed experience-dependent developmental plasticity in the mammalian auditory brainstem in an animal model. It is not known whether the human system demonstrates similar changes and whether experience with sounds composed of acoustic elements relevant to speech may alter brainstem response characteristics. We recorded brainstem responses evoked by both click and speech syllables in children between the ages of 3 and 12 years. Here, we report a neural response discrepancy in brainstem encoding of these two sounds, observed in 3- to 4-year-old children but not in school-age children. Whereas all children exhibited identical neural activity to a click, 3- to 4-year-old children displayed delayed and less synchronous onset and sustained neural response activity when elicited by speech compared with 5- to 12-year-olds. These results suggest that the human auditory system exhibits developmental plasticity, in both frequency and time domains, for sounds that are composed of acoustic elements relevant to speech. The findings are interpreted within the contexts of stimulus-related differences and experience-dependent plasticity. PMID- 18400901 TI - Absence of functional peroxisomes from mouse CNS causes dysmyelination and axon degeneration. AB - Peroxisomal metabolism is essential for normal brain development both in men and in mice. Using conditional knock-out mice, we recently showed that peroxisome deficiency in liver has a severe and persistent impact on the formation of cortex and cerebellum, whereas absence of functional peroxisomes from the CNS only causes developmental delays without obvious alteration of brain architecture. We now report that a substantial fraction of the latter Nes-Pex5 knock-out mice survive into adulthood but develop progressive motoric and coordination problems, impaired exploration, and a deficit in cognition and die before the age of 6 months. Histopathologically, both the white and gray matter of the CNS displayed a region-specific accumulation of neutral lipids, astrogliosis and microgliosis, upregulation of catalase, and scattered cell death. Nes-Pex5 knock-out mice featured a dramatic reduction of myelin staining in corpus callosum, whereas cerebellum and other white matter tracts were less affected or unchanged. This was accompanied by a depletion of alkenylphospholipids in myelin and differentially reduced immunoreactivity of myelin proteins. EM analysis revealed that myelin wrappings around axons did still form, but they showed a reduction in thickness relative to axon diameters. Remarkably, multifocal axonal damage occurred in the corpus callosum. Thereby, debris accumulated between axolemma and inner myelin surface and axons collapsed, although myelin sheaths remained present. These anomalies of myelinated axons were already present in juvenile mice but aggravated in adulthood. Together, loss of CNS peroxisomal metabolism both affects myelin sheaths and axonal integrity possibly via independent pathways. PMID- 18400900 TI - Trafficking of membrane-associated proteins to cone photoreceptor outer segments requires the chromophore 11-cis-retinal. AB - Lecithin retinol acyl transferase (LRAT) and retinal pigment epithelium protein 65 (RPE65) are key enzymes of the retinoid cycle. In Lrat(-/-) and Rpe65(-/-) mice, models of human Leber congenital amaurosis, the retinoid cycle is disrupted and 11-cis-retinal, the chromophore of visual pigments, is not produced. The Lrat(-/-) and Rpe65(-/-) retina phenotype presents with rapid sectorial cone degeneration, and the visual pigments, S-opsin and M/L-opsin, fail to traffic to cone outer segments appropriately. In contrast, rod opsin traffics normally in mutant rods. Concomitantly, guanylate cyclase 1, cone T alpha-subunit, cone phosphodiesterase 6alpha' (PDE6alpha'), and GRK1 (G-protein-coupled receptor kinase 1; opsin kinase) are not transported to Lrat(-/-) and Rpe65(-/-) cone outer segments. Aberrant localization of these membrane-associated proteins was evident at postnatal day 15, before the onset of ventral and central cone degeneration. Protein levels of cone T alpha and cone PDE6alpha' were reduced, whereas their transcript levels were unchanged, suggesting posttranslational degradation. In an Rpe65(-/-)Rho(-/-) double knock-out model, trafficking of cone pigments and membrane-associated cone phototransduction polypeptides to the outer segments proceeded normally after 11-cis-retinal administration. These results suggest that ventral and central cone opsins must be regenerated with 11-cis retinal to permit transport to the outer segments. Furthermore, the presence of 11-cis-retinal is essential for proper transport of several membrane-associated cone phototransduction polypeptides in these cones. PMID- 18400902 TI - Fear conditioning and extinction differentially modify the intrinsic excitability of infralimbic neurons. AB - Extinction of conditioned fear is an active learning process involving inhibition of fear expression. It has been proposed that fear extinction potentiates neurons in the infralimbic (IL) prefrontal cortex, but the cellular mechanisms underlying this potentiation remain unknown. It is also not known whether this potentiation occurs locally in IL neurons as opposed to IL afferents. To determine whether extinction enhances the intrinsic excitability of IL pyramidal neurons in layers II/III and V, we performed whole-cell patch-clamp recordings in slices from naive, conditioned, or conditioned-extinguished rats. We observed that conditioning depressed IL excitability compared with slices from naive animals, as evidenced by a decreased number of spikes evoked by injected current and an increase in the slow afterhyperpolarizing potential (sAHP). Extinction reversed these conditioning-induced effects. Furthermore, IL neurons from extinguished rats showed increased burst spiking compared with naive rats, which was correlated with extinction recall. These changes were specific to IL prefrontal cortex and were not observed in prelimbic prefrontal cortex. Together, these findings suggest that IL intrinsic excitability is reduced to allow for expression of conditioning memory and enhanced for expression of extinction memory through the modulation of Ca(2+)-gated K(+) channels underlying the sAHP. Inappropriate modulation of these intrinsic mechanisms may underlie anxiety disorders, characterized by exaggerated fear and deficient extinction. PMID- 18400903 TI - Tlx1 and Tlx3 coordinate specification of dorsal horn pain-modulatory peptidergic neurons. AB - The dorsal spinal cord synthesizes a variety of neuropeptides that modulate the transmission of nociceptive sensory information. Here, we used genetic fate mapping to show that Tlx3(+) spinal cord neurons and their derivatives represent a heterogeneous population of neurons, marked by partially overlapping expression of a set of neuropeptide genes, including those encoding the anti-opioid peptide cholecystokinin, pronociceptive Substance P (SP), Neurokinin B, and a late wave of somatostatin. Mutations of Tlx3 and Tlx1 result in a loss of expression of these peptide genes. Brn3a, a homeobox transcription factor, the expression of which is partly dependent on Tlx3, is required specifically for the early wave of SP expression. These studies suggest that Tlx1 and Tlx3 operate high in the regulatory hierarchy that coordinates specification of dorsal horn pain modulatory peptidergic neurons. PMID- 18400905 TI - Functional structure of the mitral cell dendritic tuft in the rat olfactory bulb. AB - The input-output transform performed by mitral cells, the principal projection neurons of the olfactory bulb, is one of the key factors in understanding olfaction. We used combined calcium and voltage imaging from the same neuron and computer modeling to investigate signal processing in the mitral cells, focusing on the glomerular dendritic tuft. The main finding was that the dendritic tuft functions as a single electrical compartment for subthreshold signals within the range of amplitudes detectable by voltage-sensitive dye recording. These evoked EPSPs had uniform characteristics throughout the glomerular tuft. The Ca(2+) transients associated with spatially uniform subthreshold synaptic potentials were comparable but not equal in amplitude in all regions. The average range of normalized amplitudes of the EPSP-driven Ca(2+) signals from different locations on dendritic branches in the glomerular tuft was relatively narrow and appeared to be independent of the dendritic surface-to-volume ratio. The computer simulations constrained by the imaging data indicated that a synchronized activation of approximately 100 synapses randomly distributed on tuft branches was sufficient to generate spatially homogenous EPSPs. This number of activated synapses is consistent with the data from anatomical studies. Furthermore, voltage attenuation of the EPSP along the primary dendrite at physiological temperature was weak compared with other cell types. In the model, weak attenuation of the EPSP along the primary dendrite could be accounted for by passive electrical properties of the mitral cell. PMID- 18400904 TI - Functional trade-offs in white matter axonal scaling. AB - The brains of large mammals have lower rates of metabolism than those of small mammals, but the functional consequences of this scaling are not well understood. An attractive target for analysis is axons, whose size, speed and energy consumption are straightforwardly related. Here we show that from shrews to whales, the composition of white matter shifts from compact, slow-conducting, and energetically expensive unmyelinated axons to large, fast-conducting, and energetically inexpensive myelinated axons. The fastest axons have conduction times of 1-5 ms across the neocortex and <1 ms from the eye to the brain, suggesting that in select sets of communicating fibers, large brains reduce transmission delays and metabolic firing costs at the expense of increased volume. Delays and potential imprecision in cross-brain conduction times are especially great in unmyelinated axons, which may transmit information via firing rate rather than precise spike timing. In neocortex, axon size distributions can account for the scaling of per-volume metabolic rate and suggest a maximum supportable firing rate, averaged across all axons, of 7 +/- 2 Hz. Axon size distributions also account for the scaling of white matter volume with respect to brain size. The heterogeneous white matter composition found in large brains thus reflects a metabolically constrained trade-off that reduces both volume and conduction time. PMID- 18400906 TI - Absence and rescue of morphine withdrawal in GIRK/Kir3 knock-out mice. AB - Although morphine induces both analgesia and dependence through mu-opioid receptors (MORs), the respective contributions of the intracellular effectors engaged by MORs remain unknown. To examine the contribution of G-protein-gated inwardly rectifying K(+) (GIRK, Kir3) channels to morphine dependence and analgesia, we quantified naloxone-precipitated withdrawal behavior and morphine analgesia using GIRK knock-out ((-/-)) mice. The morphine withdrawal syndrome was strongly attenuated, whereas morphine analgesia was mostly preserved in mice lacking both GIRK2 and GIRK3 (GIRK2/3(-/-) mice). In acute slices containing the locus ceruleus (LC) from GIRK2/3(-/-) mice, the increase in spontaneous firing typically associated with morphine withdrawal was absent. Moreover, although morphine elicited normal presynaptic inhibition in the LC, postsynaptic GIRK currents were completely abolished in GIRK2/3(-/-) mice. Altogether, these data suggested that morphine-evoked postsynaptic inhibition of the LC was required for the induction of dependence. Consistent with this hypothesis, morphine withdrawal behavior was rescued in GIRK2/3(-/-) mice by ablation of adrenergic fibers using the neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine. Our data suggest that inhibition of adrenergic tone is required for the induction of dependence, and that channels containing GIRK2 and GIRK3 serve as an inhibitory gate. PMID- 18400907 TI - Functional asymmetries in visual pathways carrying S-cone signals in macaque. AB - In the lateral geniculate nucleus of macaque, we recorded from neurons with substantial input from S-cones and found that, on several important dimensions, the properties of neurons that receive inhibitory input from S-cones ("S-") are quite unlike those of neurons that receive excitatory input from S-cones ("S+"). First, the organization of chromatic inputs differs substantially: in S+ cells, S cone signals were usually opposed by those of L- and M-cones; in S- cells, signals from L-cones were usually opposed to those of S- and M-cones. Second, to pure S-cone modulation, S+ cells are twice as sensitive as S- cells, but S- cells were much more susceptible to contrast adaptation. Third, in S- cells but not S+ cells, the spatial frequency resolution for achromatic modulation was often greater, the tuning curve and more bandpass, than that for S-cone modulation. Along the dimensions on which we measured, the properties of the S+ cells were relatively tightly clustered, suggesting a homogenous class. Although the chromatic properties of S- cells are heterogeneous, the distribution of their tuning along other stimulus dimensions does not suggest multiple subtypes. PMID- 18400909 TI - The hippocamposeptal pathway generates rhythmic firing of GABAergic neurons in the medial septum and diagonal bands: an investigation using a complete septohippocampal preparation in vitro. AB - The medial septum diagonal band area (MS/DB) projects to the hippocampus through the fornix/fimbria pathway and is implicated in generating hippocampal theta oscillations. The hippocampus also projects back to the MS/DB, but very little is known functionally about this input. Here, we investigated the physiological role of hippocamposeptal feedback to the MS/DB in a complete in vitro septohippocampal preparation containing the intact interconnecting fornix/fimbria pathway. We demonstrated that carbachol-induced rhythmic theta-like hippocampal oscillations recorded extracellularly were synchronized with powerful rhythmic IPSPs in whole cell recorded MS/DB neurons. Interestingly, we found that these IPSPs evoked rebound spiking in GABAergic MS/DB neurons. In contrast, putative cholinergic and glutamatergic MS/DB neurons responded only weakly with rebound spiking and, as a result, were mostly silent during theta-like oscillations. We next determined the mechanism underlying the rebound spiking that followed the IPSPs in MS/DB GABAergic neurons using phasic electrical stimulation of the fornix/fimbria pathway. We demonstrate that the increased rebound spiking was attributable to the activation of I(h) current, because it was significantly reduced by low concentrations of the I(h) antagonist ZD7288 [4-(N-ethyl-N-phenylamino)-1,2 dimethyl-6-(methylamino) pyridinium chloride]. Together, these results suggest that rhythmical activity in hippocampus is transferred to the MS/DB and can preferentially phase the spiking of GABAergic MS/DB neurons because of their significant expression of I(h) currents. Our data demonstrate that hippocamposeptal inhibition facilitates theta rhythmic discharges in MS/DB GABAergic neurons while favoring the inhibition of most ACh and glutamate neurons. PMID- 18400908 TI - A causal role for brain-derived neurotrophic factor in the homeostatic regulation of sleep. AB - Slow-wave activity (SWA), the EEG power between 0.5 and 4 Hz during non-rapid eye movement (NREM) sleep, is one of the best characterized markers of sleep need, because it increases as a function of preceding waking duration and decreases during sleep, but the underlying mechanisms remain unknown. We hypothesized that SWA is high at sleep onset because it reflects the occurrence, during the previous waking period, of widespread synaptic potentiation in cortical and subcortical areas. Consistent with this hypothesis, we recently showed that the more rats explore, the stronger is the cortical expression of BDNF during wakefulness, and the larger is the increase in SWA during the subsequent sleep period. There is compelling evidence that BDNF plays a causal role in synaptic potentiation, and exogenous application of BDNF in vivo is sufficient to induce long-term increases in synaptic strength. We therefore performed cortical unilateral microinjections of BDNF in awake rats and measured SWA during the subsequent sleep period. SWA during NREM sleep was higher in the injected hemisphere relative to the contralateral one. The effect was reversible within 2 h, and did not occur during wakefulness or rapid eye movement sleep. Asymmetries in NREM SWA did not occur after vehicle injections. Furthermore, microinjections, during wakefulness, of a polyclonal anti-BDNF antibody or K252a, an inhibitor of BDNF TrkB receptors, led to a local SWA decrease during the following sleep period. These effects were also reversible and specific for NREM sleep. These results show a causal link between BDNF expression during wakefulness and subsequent sleep regulation. PMID- 18400910 TI - Field evaluation of a rapid immunochromatographic assay for detection of Trypanosoma cruzi infection by use of whole blood. AB - Laboratory and clinical diagnostic classification of seropositive individuals, followed by treatment and supportive therapy, is an established component of Chagas' disease control in areas where this disease is endemic. However, most Chagas' disease patients live in remote areas where neither equipped laboratories nor skilled human resources are widely available. Employing a rapid diagnostic test (RDT), when using whole blood samples, is the best option for Chagas' disease control. A high sensitivity and specificity for the Chagas Stat-Pak RDT (Chembio Diagnostic Systems, Inc., Medford, NY) has been reported for assays using serum and plasma, but its validity for the detection of antibodies to Trypanosoma cruzi infection in whole blood is unknown. This cross-sectional study measured the sensitivity and specificity of the Chagas Stat-Pak with whole blood, using conventional serological assays for comparison. The interobserver reliability in the interpretation of the Chagas Stat-Pak results and "ease-of use" criterion needed to perform the Chagas Stat-Pak and conventional assays were also measured. The Chagas Stat-Pak yielded a high specificity (99.0%, 95% confidence interval [CI] = 98.4 to 99.4%) but a relatively low sensitivity (93.4%, 95% CI = 87.4 to 97.1%). The interobserver reliability was excellent (kappa [n = 1,913] = 0.999, P < 0.0001), and the quantified ease-of-use criterion suggested that the RDT is simple to perform. Despite the attributes of the Chagas Stat-Pak, it is not an ideal diagnostic test for the population investigated in the present study due to its relatively low sensitivity and high cost. The RDT manufacturer is called upon to improve the test if the international community hopes to make progress in controlling Chagas infections in areas where this disease is endemic. PMID- 18400911 TI - In vitro replication of varicella-zoster virus in human retinal pigment epithelial cells. AB - Here we describe for the first time the productive in vitro infection of human retinal pigment epithelial cells by varicella-zoster virus (VZV), resulting in a typical cytopathic effect (CPE) that is characterized by enlarged cells with increased granularity. Depending on the CPE dissemination, high titers of up to 1.6 x 10(6) PFU of cell-free and cryostable VZV/ml can be recovered. PMID- 18400912 TI - Comparison of sensitivities of two commercial gamma interferon release assays for pulmonary tuberculosis. AB - There are few head-to-head comparisons of the commercial gamma interferon release assays (GIRAs). We compared the performance of the T-SPOT.TB and QuantiFERON-TB Gold In-Tube (QFT-IT) assays in patients with culture-proven pulmonary tuberculosis. Blood was drawn for both assays within 14 days of starting antituberculosis treatment. The QFT-IT indeterminate rate was 3.5%; the T-SPOT.TB failure rate was 1.4%. There was poor agreement between the GIRAs (kappa = 0.257) among the 270 patients with valid results for both tests. The sensitivities of the T-SPOT.TB and QFT-IT assays were 94.1 and 83.0%, respectively, with a significant difference in the performance of the assays (P = 0.001 [McNemar test]). Factors independently associated with indeterminate QFT-IT results were an age of >or=60 years (odds ratio [OR] 11.18, 95% confidence interval [CI] = 1.841 to 67.823, P = 0.009), female sex (OR = 7.47, 95% CI = 1.517 to 36.733, P = 0.013) and non-Chinese (i.e., Indian or Malay) race (OR = 7.89, 95% CI = 1.585 to 39.267, P = 0.012). The QFT-IT assay was significantly less sensitive in patients >or=60 years old (OR = 0.41, 95% CI = 0.181 to 0.918, P = 0.030) and in Indian compared to Chinese patients (OR = 0.27, 95% CI = 0.073 to 0.990, P = 0.048). The T-SPOT.TB assay was significantly less sensitive in Malay (OR = 0.23, 95% CI = 0.063 to 0.815, P = 0.023) and Indian patients (OR = 0.09, 95% CI = 0.017 to 0.429, P = 0.003) compared to Chinese patients. The performance of both assays was not significantly altered in diabetics. The diminished sensitivity of the GIRAs in persons of Malay and Indian race merits further study. PMID- 18400913 TI - Evaluation of Versant hepatitis C virus genotype assay (LiPA) 2.0. AB - Hepatitis C virus (HCV) genotyping is a tool used to optimize antiviral treatment regimens. The newly developed Versant HCV genotype assay (LiPA) 2.0 uses sequence information from both the 5' untranslated region and the core region, allowing distinction between HCV genotype 1 and subtypes c to l of genotype 6 and between subtypes a and b of genotype 1. HCV-positive samples were genotyped manually using the Versant HCV genotype assay (LiPA) 2.0 system according to the manufacturer's instructions. For the comparison study, Versant HCV genotype assay (LiPA) 1.0 was used. In this study, 99.7% of the samples could be amplified, the genotype of 96.0% of samples could be determined, and the agreement with the reference method was 99.4% when a genotype was determined. The reproducibility study showed no significant differences in performance across sites (P = 0.43) or across lots (P = 0.88). In the comparison study, 13 samples that were uninterpretable or incorrectly genotyped with Versant HCV genotype assay (LiPA) 1.0 were correctly genotyped by Versant HCV genotype assay (LiPA) 2.0. Versant HCV genotype assay (LiPA) 2.0 is a sensitive, accurate, and reliable assay for HCV genotyping. The inclusion of the core region probes in Versant HCV genotype assay (LiPA) 2.0 results in a genotyping success rate higher than that of the current Versant HCV genotype assay (LiPA) 1.0. PMID- 18400915 TI - Genetic differentiation of Escherichia coli O157:H7 clades associated with human disease by real-time PCR. AB - The rapid and accurate identification of Escherichia coli O157:H7 strains is central to reducing the impact of outbreaks. A real-time PCR-based approach to differentiating major outbreak lineages of O157 with novel single-nucleotide polymorphisms is described. The utility of this method is in detection of hypervirulent strains in cases of clinical disease. PMID- 18400914 TI - Quantitative real-time PCR is not more sensitive than "conventional" PCR. PMID- 18400916 TI - Development and evaluation of a novel multiplex PCR technology for molecular differential detection of bacterial respiratory disease pathogens. AB - The ResPlex I assay (Qiagen) was designed to amplify and detect DNA of six bacterial respiratory pathogens. This assay was compared with real-time PCR assays based upon the same target sequences for the ability detect the target bacteria by use of both stock strains and specimens from respiratory disease patients. The ResPlex I assay is somewhat less sensitive than real-time PCR assays but offers the advantage of multiple assays in a single reaction. PMID- 18400917 TI - Comparison of immunoblotting, calculation of the Goldmann-Witmer coefficient, and real-time PCR using aqueous humor samples for diagnosis of ocular toxoplasmosis. AB - We compared three biological methods for the diagnosis of ocular toxoplasmosis (OT). Paired aqueous humor and serum samples from 34 patients with OT and from 76 patients with other ocular disorders were analyzed by three methods: immunoblotting or Western blotting (WB), the calculation of the Goldmann-Witmer coefficient (GWC), and PCR. WB and GWC each revealed the intraocular production of specific anti-Toxoplasma immunoglobulin G in 81% of samples (30 of 37). PCR detected toxoplasmic DNA in 38% of samples (13 of 34). Nine of the 13 PCR positive patients were immunocompetent. Combining the techniques significantly improved the diagnostic sensitivity, to 92% for the GWC-WB combination, 90% for the WB-PCR combination, and 93% for the GWC-PCR combination. The combination of all three techniques improved the sensitivity to 97%. PMID- 18400918 TI - Caspofungin disk diffusion breakpoints and quality control. AB - Interpretive disk diffusion breakpoints for caspofungin are proposed by evaluating 762 isolates of Candida spp., representing 10 different species obtained as part of the caspofungin clinical trials. Standardized broth microdilution reference tests were compared to the zone diameters observed with 5 microg caspofungin disks produced by two different disk manufacturers. Disk diffusion breakpoints of >or=11 mm for susceptible are proposed. Compared to results from MIC testing, these zone diameters produced error rates that were or=99% sequence similarity); 11 further isolates gave ambiguous results at this threshold and were rated as identified to the genus level only. Ten isolates were identified to the genus level (>or=97% similarity); and two isolates had similarity values below this threshold, were counted as not identified, and were excluded from further analysis. MALDI-TOF MS identified 67 of the 78 isolates (85.9%) included, in agreement with the results of the reference method; 9 were misidentified and 2 were unidentified. The identities of 10 randomly selected strains were 100% correct when three different mass spectrometers and four different cultivation media were used. Thus, MALDI-TOF MS based species identification of nonfermenting bacteria provided accurate and reproducible results within 10 min without any substantial costs for consumables. PMID- 18400922 TI - The brain's default network: anatomy, function, and relevance to disease. AB - Thirty years of brain imaging research has converged to define the brain's default network-a novel and only recently appreciated brain system that participates in internal modes of cognition. Here we synthesize past observations to provide strong evidence that the default network is a specific, anatomically defined brain system preferentially active when individuals are not focused on the external environment. Analysis of connectional anatomy in the monkey supports the presence of an interconnected brain system. Providing insight into function, the default network is active when individuals are engaged in internally focused tasks including autobiographical memory retrieval, envisioning the future, and conceiving the perspectives of others. Probing the functional anatomy of the network in detail reveals that it is best understood as multiple interacting subsystems. The medial temporal lobe subsystem provides information from prior experiences in the form of memories and associations that are the building blocks of mental simulation. The medial prefrontal subsystem facilitates the flexible use of this information during the construction of self-relevant mental simulations. These two subsystems converge on important nodes of integration including the posterior cingulate cortex. The implications of these functional and anatomical observations are discussed in relation to possible adaptive roles of the default network for using past experiences to plan for the future, navigate social interactions, and maximize the utility of moments when we are not otherwise engaged by the external world. We conclude by discussing the relevance of the default network for understanding mental disorders including autism, schizophrenia, and Alzheimer's disease. PMID- 18400923 TI - Episodic simulation of future events: concepts, data, and applications. AB - This article focuses on the neural and cognitive processes that support imagining or simulating future events, a topic that has recently emerged in the forefront of cognitive neuroscience. We begin by considering concepts of simulation from a number of areas of psychology and cognitive neuroscience in order to place our use of the term in a broader context. We then review neuroimaging, neuropsychological, and cognitive studies that have examined future-event simulation and its relation to episodic memory. This research supports the idea that simulating possible future events depends on much of the same neural machinery, referred to here as a core network, as does remembering past events. After discussing several theoretical accounts of the data, we consider applications of work on episodic simulation for research concerning clinical populations suffering from anxiety or depression. Finally, we consider other aspects of future-oriented thinking that we think are related to episodic simulation, including planning, prediction, and remembering intentions. These processes together comprise what we have termed "the prospective brain," whose primary function is to use past experiences to anticipate future events. PMID- 18400924 TI - Generalization and differentiation in semantic memory: insights from semantic dementia. AB - According to many theories, semantic representations reflect the parallel activation of information coded across a distributed set of modality-specific association brain cortices. This view is challenged by the neurodegenerative condition known as semantic dementia (SD), in which relatively circumscribed, bilateral atrophy of the anterior temporal lobes results in selective degradation of core semantic knowledge, affecting all types of concept, irrespective of the modality of testing. Research on SD suggests a major revision in our understanding of the neural basis of semantic memory. Specifically, it is proposed that the anterior temporal lobes form amodal semantic representations through the distillation of the multimodal information that is projected to this region from the modality-specific association cortices. Although cross-indexing of modality-specific information could be achieved by a web of direct connections between pairs of these regions, amodal semantic representations enable semantic generalization and inference on the basis of conceptual structure rather than modality-specific features. As expected from this hypothesis, SD is characterized by impaired semantic generalization, both clinically and in formal assessment. The article describes a comprehensive array of under- and overgeneralization errors by patients with SD when engaged in receptive and expressive verbal and nonverbal tasks and everyday behaviors. PMID- 18400925 TI - Spatial cognition and the brain. AB - Recent advances in the understanding of spatial cognition are reviewed, focusing on memory for locations in large-scale space and on those advances inspired by single-unit recording and lesion studies in animals. Spatial memory appears to be supported by multiple parallel representations, including egocentric and allocentric representations, and those updated to accommodate self-motion. The effects of these representations can be dissociated behaviorally, developmentally, and in terms of their neural bases. It is now becoming possible to construct a mechanistic neural-level model of at least some aspects of spatial memory and imagery, with the hippocampus and medial temporal lobe providing allocentric environmental representations, the parietal lobe egocentric representations, and the retrosplenial cortex and parieto-occipital sulcus allowing both types of representation to interact. Insights from this model include a common mechanism for the construction of spatial scenes in the service of both imagery and episodic retrieval and a role for the remainder of Papez's circuit in orienting the viewpoint used. In addition, it appears that hippocampal and striatal systems process different aspects of environmental layout (boundaries and local landmarks, respectively) and do so using different learning rules (incidental learning and associative reinforcement, respectively). PMID- 18400926 TI - Multisensory-based approach to the recovery of unisensory deficit. AB - This chapter reviews several highly convergent behavioral findings that provide strong evidence for the existence of multimodal integration systems subserving spatial representation in humans. These systems generally function through the multisensory coding of visuoauditory and visuotactile events but vary in their specific functional and anatomical characteristics. The chapter will also consider the adaptive advantages of multisensory integration systems; these systems might modulate the level of activation in cortical areas in short- and long-term ways, thereby providing a mechanism for permanent recovery from sensory and spatial deficits. PMID- 18400928 TI - Cognitive neuroscience of aging. AB - The number of reports on the cognitive neuroscience of aging has increased in recent years, and most of these studies have found many similarities in the patterns of activity in young and old adults, indicating that basic neural mechanisms are maintained into older age. Despite these overall similarities, older adults often have less activity in some regions, such as medial temporal areas during memory processing and visual regions across a variety of cognitive domains. It seems clear that age reductions in cognitive function can be tied, at least in part, to these reductions in brain activity. On the other hand, older adults typically also overrecruit some brain areas, mainly the ventral or dorsal prefrontal cortex during memory tasks, as well as both the frontal and parietal regions during tasks engaging cognitive control processes, such as attention. Sometimes this overrecruitment appears to be in response to altered function in other brain regions and is often seen in those older adults who perform better on the task at hand. These findings have provided rather convincing support for the idea that overrecruitment can be compensatory in the elderly. Nevertheless, not all age increases can be interpreted as compensatory, and some are more indicative of neural inefficiency. The challenge facing future research will be to understand the task conditions that promote compensation in older adults, the role of the various brain areas in aiding cognitive function, and how these compensatory mechanisms can be elicited to enhance quality of life in the elderly. PMID- 18400927 TI - The adolescent brain. AB - Adolescence is a developmental period characterized by suboptimal decisions and actions that are associated with an increased incidence of unintentional injuries, violence, substance abuse, unintended pregnancy, and sexually transmitted diseases. Traditional neurobiological and cognitive explanations for adolescent behavior have failed to account for the nonlinear changes in behavior observed during adolescence, relative to both childhood and adulthood. This review provides a biologically plausible model of the neural mechanisms underlying these nonlinear changes in behavior. We provide evidence from recent human brain imaging and animal studies that there is a heightened responsiveness to incentives and socioemotional contexts during this time, when impulse control is still relatively immature. These findings suggest differential development of bottom-up limbic systems, implicated in incentive and emotional processing, to top-down control systems during adolescence as compared to childhood and adulthood. This developmental pattern may be exacerbated in those adolescents prone to emotional reactivity, increasing the likelihood of poor outcomes. PMID- 18400929 TI - Can neurological evidence help courts assess criminal responsibility? Lessons from law and neuroscience. AB - Can neurological evidence help courts assess criminal responsibility? To answer this question, we must first specify legal criteria for criminal responsibility and then ask how neurological findings can be used to determine whether particular defendants meet those criteria. Cognitive neuroscience may speak to at least two familiar conditions of criminal responsibility: intention and sanity. Functional neuroimaging studies in motor planning, awareness of actions, agency, social contract reasoning, and theory of mind, among others, have recently targeted a small assortment of brain networks thought to be instrumental in such determinations. Advances in each of these areas bring specificity to the problems underlying the application of neuroscience to criminal law. PMID- 18400930 TI - The neural basis of moral cognition: sentiments, concepts, and values. AB - The human moral nature has perplexed laymen and academics for millennia. Recent developments in cognitive neuroscience are opening new venues for unveiling the complex psychological and neurobiological mechanisms underling human morality and its impairments. Here we review these lines of evidence and key topics of debate and explain why investigating the mechanisms of cognition-emotion interaction and of the neural bases of moral sentiments and values will be critical for our understanding of the human moral mind. PMID- 18400931 TI - Intention, choice, and the medial frontal cortex. AB - The medial frontal cortex (MFC) has been identified with voluntary action selection. Recent evidence suggests that there are three principal ways in which the MFC is an essential part of the neural circuit for voluntary action selection. First, the MFC represents the reinforcement values of actions and is concerned with the updating of those action values. Because it is particularly concerned with the rate at which action values should be updated, it mediates the influence that the past reinforcement history has over the next choice that is made and it may determine the learning rate. The MFC's representation of action value does not just reflect the potential reward associations of an action but instead represents both the reward and effort costs that are intrinsic to the action. Second, the MFC is important when an exploratory action is generated in order to obtain more information about action values and the environment. Third, the MFC is critical when conflicting information in the immediate environment instructs more than one possible response. In such situations the MFC exerts an influence over how actions will be chosen by other motor regions of the brain. PMID- 18400932 TI - Evaluating faces on trustworthiness: an extension of systems for recognition of emotions signaling approach/avoidance behaviors. AB - People routinely make various trait judgments from facial appearance, and such judgments affect important social outcomes. These judgments are highly correlated with each other, reflecting the fact that valence evaluation permeates trait judgments from faces. Trustworthiness judgments best approximate this evaluation, consistent with evidence about the involvement of the amygdala in the implicit evaluation of face trustworthiness. Based on computer modeling and behavioral experiments, I argue that face evaluation is an extension of functionally adaptive systems for understanding the communicative meaning of emotional expressions. Specifically, in the absence of diagnostic emotional cues, trustworthiness judgments are an attempt to infer behavioral intentions signaling approach/avoidance behaviors. Correspondingly, these judgments are derived from facial features that resemble emotional expressions signaling such behaviors: happiness and anger for the positive and negative ends of the trustworthiness continuum, respectively. The emotion overgeneralization hypothesis can explain highly efficient but not necessarily accurate trait judgments from faces, a pattern that appears puzzling from an evolutionary point of view and also generates novel predictions about brain responses to faces. Specifically, this hypothesis predicts a nonlinear response in the amygdala to face trustworthiness, confirmed in functional magnetic resonance imaging (fMRI) studies, and dissociations between processing of facial identity and face evaluation, confirmed in studies with developmental prosopagnosics. I conclude with some methodological implications for the study of face evaluation, focusing on the advantages of formally modeling representation of faces on social dimensions. PMID- 18400933 TI - Disorders of consciousness. AB - The vegetative state and other so-called disorders of consciousness present some of the most significant practical and ethical challenges in modern medicine. It is extremely difficult to assess residual cognitive function in these patients because their movements may be minimal or inconsistent, or because no cognitive output may be possible. In recent years, behavioral and neuroimaging techniques developed within the cognitive neurosciences have provided a number of new approaches for investigating these disorders, leading to significant advances in current understanding. In several cases, residual cognitive function and even conscious awareness have been demonstrated in patients who are assumed to be vegetative yet retain cognitive abilities that have evaded detection using standard clinical methods. In this article, I review these data, focusing primarily on the vegetative and minimally conscious states. PMID- 18400934 TI - The neural correlates of consciousness: an update. AB - This review examines recent advances in the study of brain correlates of consciousness. First, we briefly discuss some useful distinctions between consciousness and other brain functions. We then examine what has been learned by studying global changes in the level of consciousness, such as sleep, anesthesia, and seizures. Next we consider some of the most common paradigms used to study the neural correlates for specific conscious percepts and examine what recent findings say about the role of different brain regions in giving rise to consciousness for that percept. Then we discuss dynamic aspects of neural activity, such as sustained versus phasic activity, feedforward versus reentrant activity, and the role of neural synchronization. Finally, we briefly consider how a theoretical analysis of the fundamental properties of consciousness can usefully complement neurobiological studies. PMID- 18400935 TI - Differential regulation of the expression of two high-affinity sulfate transporters, SULTR1.1 and SULTR1.2, in Arabidopsis. AB - The molecular mechanisms regulating the initial uptake of inorganic sulfate in plants are still largely unknown. The current model for the regulation of sulfate uptake and assimilation attributes positive and negative regulatory roles to O acetyl-serine (O-acetyl-Ser) and glutathione, respectively. This model seems to suffer from exceptions and it has not yet been clearly validated whether intracellular O-acetyl-Ser and glutathione levels have impacts on regulation. The transcript level of the two high-affinity sulfate transporters SULTR1.1 and SULTR1.2 responsible for sulfate uptake from the soil solution was compared to the intracellular contents of O-acetyl-Ser, glutathione, and sulfate in roots of plants submitted to a wide diversity of experimental conditions. SULTR1.1 and SULTR1.2 were differentially expressed and neither of the genes was regulated in accordance with the current model. The SULTR1.1 transcript level was mainly altered in response to the sulfur-related treatments. Split-root experiments show that the expression of SULTR1.1 is locally regulated in response to sulfate starvation. In contrast, accumulation of SULTR1.2 transcripts appeared to be mainly related to metabolic demand and is controlled by photoperiod. On the basis of the new molecular insights provided in this study, we suggest that the expression of the two transporters depends on different regulatory networks. We hypothesize that interplay between SULTR1.1 and SULTR1.2 transporters could be an important mechanism to regulate sulfate content in the roots. PMID- 18400936 TI - Role of swollenin, an expansin-like protein from Trichoderma, in plant root colonization. AB - Swollenin, a protein first characterized in the saprophytic fungus Trichoderma reesei, contains an N-terminal carbohydrate-binding module family 1 domain (CBD) with cellulose-binding function and a C-terminal expansin-like domain. This protein was identified by liquid chromatography-mass spectrometry among many other cellulolytic proteins secreted in the coculture hydroponics medium of cucumber (Cucumis sativus) seedlings and Trichoderma asperellum, a well-known biocontrol agent and inducer of plant defense responses. The swollenin gene was isolated and its coding region was overexpressed in the same strain under the control of the constitutive pki1 promoter. Trichoderma transformants showed a remarkably increased ability to colonize cucumber roots within 6 h after inoculation. On the other hand, overexpressors of a truncated swollenin sequence bearing a 36-amino acid deletion of the CBD did not differ from the wild type, showing in vivo that this domain is necessary for full protein activity. Root colonization rates were reduced in transformants silenced in swollenin gene expression. A synthetic 36-mer swollenin CBD peptide was shown to be capable of stimulating local defense responses in cucumber roots and leaves and to afford local protection toward Botrytis cinerea and Pseudomonas syringae pv lachrymans infection. This indicates that the CBD domain might be recognized by the plant as a microbe-associated molecular pattern in the Trichoderma-plant interaction. PMID- 18400937 TI - The pentratricopeptide repeat protein DELAYED GREENING1 is involved in the regulation of early chloroplast development and chloroplast gene expression in Arabidopsis. AB - An Arabidopsis (Arabidopsis thaliana) mutant that exhibited a delayed greening phenotype (dg1) was isolated from a population of activation-tagged Arabidopsis lines. Young, inner leaves of dg1 mutants were initially very pale, but gradually greened and mature outer leaves, more than 3 weeks old, appeared similar to those of wild-type plants. Sequence and transcription analyses showed that DG1 encodes a chloroplast protein consisting of eight pentratricopeptide repeat domains and that its expression depends on both light and developmental status. In addition, analysis of the transcript profiles of chloroplast genes revealed that plastid encoded polymerase-dependent transcript levels were markedly reduced, while nucleus-encoded polymerase-dependent transcript levels were increased, in dg1 mutants. Thus, DG1 is probably involved in the regulation of plastid-encoded polymerase-dependent chloroplast gene expression during early stages of chloroplast development. PMID- 18400938 TI - Impact of activated vitamin D and race on survival among hemodialysis patients. AB - Contrary to most examples of disparities in health outcomes, black patients have improved survival compared with white patients after initiating hemodialysis. Understanding potential explanations for this observation may have important clinical implications for minorities in general. This study tested the hypothesis that greater use of activated vitamin D therapy accounts for the survival advantage observed in black and Hispanic patients on hemodialysis. In a prospective cohort of non-Hispanic white (n = 5110), Hispanic white (n = 979), and black (n = 3214) incident hemodialysis patients, higher parathyroid hormone levels at baseline were the primary determinant of prescribing activated vitamin D therapy. Median parathyroid hormone was highest among black patients, who were most likely to receive activated vitamin D and at the highest dosage. One-year mortality was lower in black and Hispanic patients compared with white patients (16 and 16 versus 23%; P < 0.01), but there was significant interaction between race and ethnicity, activated vitamin D therapy, and survival. In multivariable analyses of patients treated with activated vitamin D, black patients had 16% lower mortality compared with white patients, but the difference was lost when adjusted for vitamin D dosage. In contrast, untreated black patients had 35% higher mortality compared with untreated white patients, an association that persisted in several sensitivity analyses. In conclusion, therapy with activated vitamin D may be one potential explanation for the racial differences in survival among hemodialysis patients. Further studies should determine whether treatment differences based on biologic differences contribute to disparities in other conditions. PMID- 18400939 TI - The effect of raloxifene treatment in postmenopausal women with CKD. AB - It is unknown whether treatment for osteoporosis with raloxifene is safe or effective in those with chronic kidney disease (CKD). With data from a multicenter, randomized, placebo-controlled trial of 7705 postmenopausal women with osteoporosis, the effect of raloxifene on rate of change of bone mineral density (BMD), incidence of fractures, and adverse events by stage of CKD was examined over 3 yr. Baseline serum creatinine values were available for 7316 women, and these values were used to assign a category of creatinine clearance (CrCl) using the Cockcroft-Gault formula (CrCl < 45, 45 to 59, and > or = 60 ml/min). BMD was measured at baseline and annually by dual x-ray absorptiometry. Within the placebo group, lower baseline CrCl was associated with a trend for higher annual losses of BMD at the femoral neck; however, within the raloxifene group, lower baseline CrCl was associated with greater increases in femoral neck BMD. This interaction between category of CrCl and treatment assignment was significant for rate of change of BMD at the hip. Irrespective of kidney function, raloxifene treatment was associated with a greater increase in spine BMD, a reduction in vertebral fractures, and no effect on nonvertebral fractures compared with placebo. Within each category of kidney function, adverse events were similar between the raloxifene and placebo groups. In conclusion, raloxifene increases BMD at both the hip and the spine and reduces the risk for vertebral fractures among individuals with CKD. The effect ofraloxifene on hip BMD is greater among those with mild to moderate CKD. PMID- 18400940 TI - Autonomic renal denervation ameliorates experimental glomerulonephritis. AB - Increasing evidence indicates that inflammation of visceral organs is significantly affected by the autonomic nervous system. Such neuroimmune interactions have not been studied in the kidney. Here, we show that the rat kidney is innervated by both tyrosine hydroxylase-positive sympathetic efferent nerve fibers and calcitonin gene-related peptide-positive primary afferent nerve fibers, both of which are found in proximity to macrophages and dendritic cells. Complete surgical bilateral renal denervation was performed 2 d before glomerulonephritis was induced by injecting the monoclonal anti-Thy-1.1 antibody OX-7. Denervation significantly reduced albuminuria, mesangiolysis, formation of microaneurysms, deposition of glomerular collagen IV, and expression of TGF-beta compared with sham-operated controls. Accordingly, inflammation, identified by accumulation of interstitial macrophages and renal expression of TNF-alpha, and mesangial cell proliferation were significantly reduced. These findings indicate that autonomic renal denervation ameliorates and, by inference, innervation exacerbates acute inflammation in the kidney; therefore, neurotransmitters or neuropeptides and their receptors might represent novel targets for the treatment of acute glomerulonephritis. PMID- 18400941 TI - Pitfalls of measuring total blood calcium in patients with CKD. AB - Disorders of mineral and bone metabolism are prevalent in patients with chronic kidney disease (CKD). The recent National Kidney Foundation Kidney Disease Outcomes Quality Initiative (K/DOQI) guidelines recommend that blood calcium (Ca) be regularly measured in patients with stages 3 to 5 CKD. The Kidney Disease: Improving Global Outcomes (KDIGO) position states that the measurement of ionized Ca (iCa) is preferred and that if total Ca (tCa) concentration is used instead, then it should be adjusted in the setting of hypoalbuminemia. In 691 consecutive patients with stages 3 to 5 CKD, we compared the ability of noncorrected and albumin-corrected tCa concentration to identify low, normal, or high iCa concentration. The agreement between noncorrected or albumin-corrected tCa and iCa was only fair. The risk for underestimating ionized calcium was independently increased by a low total CO(2) concentration when either noncorrected or albumin corrected Ca was used and by a low albumin concentration only when noncorrected tCa was used. The risk for overestimating iCa was increased by a low albumin concentration only when albumin-corrected Ca was used. In conclusion, albumin corrected tCa does not predict iCa better than noncorrected tCa. Moreover, both noncorrected and albumin-corrected tCa concentrations poorly predict hypo- or hypercalcemia in patients with CKD. PMID- 18400942 TI - R-Spondin family members regulate the Wnt pathway by a common mechanism. AB - The R-Spondin (RSpo) family of secreted proteins is implicated in the activation of the Wnt signaling pathway. Despite the high structural homology between the four members, expression patterns and phenotypes in knockout mice have demonstrated striking differences. Here we dissected and compared the molecular and cellular function of all RSpo family members. Although all four RSpo proteins activate the canonical Wnt pathway, RSpo2 and 3 are more potent than RSpo1, whereas RSpo4 is relatively inactive. All RSpo members require Wnt ligands and LRP6 for activity and amplify signaling of Wnt3A, Wnt1, and Wnt7A, suggesting that RSpo proteins are general regulators of canonical Wnt signaling. Like RSpo1, RSpo2-4 antagonize DKK1 activity by interfering with DKK1 mediated LRP6 and Kremen association. Analysis of RSpo deletion mutants indicates that the cysteine rich furin domains are sufficient and essential for the amplification of Wnt signaling and inhibition of DKK1, suggesting that Wnt amplification by RSpo proteins may be a direct consequence of DKK1 inhibition. Together, these findings indicate that RSpo proteins modulate the Wnt pathway by a common mechanism and suggest that coexpression with specific Wnt ligands and DKK1 may determine their biological specificity in vivo. PMID- 18400943 TI - Cross-talk between fibroblast growth factor and bone morphogenetic proteins regulates gap junction-mediated intercellular communication in lens cells. AB - Homeostasis in the lens is dependent on an extensive network of cell-to-cell gap junctional channels. Gap junction-mediated intercellular coupling (GJIC) is higher in the equatorial region of the lens than at either pole, an asymmetry believed essential for lens transparency. Primary cultures of embryonic chick lens epithelial cells up-regulate GJIC in response to purified fibroblast growth factor (FGF)1/2 or to medium conditioned by vitreous bodies, the major reservoir of factors (including FGF) for the lens equator. We show that purified bone morphogenetic protein (BMP)2, -4, and -7 also up-regulate GJIC in these cultures. BMP2, -4, or both are present in vitreous body conditioned medium, and BMP4 and 7 are endogenously expressed by lens cells. Remarkably, lens-derived BMP signaling is required for up-regulation of GJIC by purified FGF, and sufficient for up-regulation by vitreous humor. This is the first demonstration of an obligatory interaction between FGF and BMPs in postplacode lens cells, and of a role for FGF/BMP cross-talk in regulating GJIC in any cell type. Our results support a model in which the angular gradient in GJIC in the lens, and thus proper lens function, is dependent on signaling between the FGF and BMP pathways. PMID- 18400944 TI - Mitochondrial protein import motor: differential role of Tim44 in the recruitment of Pam17 and J-complex to the presequence translocase. AB - The presequence translocase of the mitochondrial inner membrane (TIM23 complex) mediates the import of preproteins with amino-terminal presequences. To drive matrix translocation the TIM23 complex recruits the presequence translocase associated motor (PAM) with the matrix heat shock protein 70 (mtHsp70) as central subunit. Activity and localization of mtHsp70 are regulated by four membrane associated cochaperones: the adaptor protein Tim44, the stimulatory J-complex Pam18/Pam16, and Pam17. It has been proposed that Tim44 serves as molecular platform to localize mtHsp70 and the J-complex at the TIM23 complex, but it is unknown how Pam17 interacts with the translocase. We generated conditional tim44 yeast mutants and selected a mutant allele, which differentially affects the association of PAM modules with TIM23. In tim44-804 mitochondria, the interaction of the J-complex with the TIM23 complex is impaired, whereas unexpectedly the binding of Pam17 is increased. Pam17 interacts with the channel protein Tim23, revealing a new interaction site between TIM23 and PAM. Thus, the motor PAM is composed of functional modules that bind to different sites of the translocase. We suggest that Tim44 is not simply a scaffold for binding of motor subunits but plays a differential role in the recruitment of PAM modules to the inner membrane translocase. PMID- 18400945 TI - A novel group of glutaredoxins in the cis-Golgi critical for oxidative stress resistance. AB - Glutaredoxins represent a ubiquitous family of proteins that catalyze the reduction of disulfide bonds in their substrate proteins by use of reduced glutathione. In an attempt to identify the full complement of glutaredoxins in baker's yeast, we found three so-far uncharacterized glutaredoxin-like proteins that we named Grx6, Grx7, and Grx8. Grx6 and Grx7 represent closely related monothiol glutaredoxins that are synthesized with N-terminal signal sequences. Both proteins are located in the cis-Golgi, thereby representing the first glutaredoxins found in a compartment of the secretory pathway. In contrast to formerly described monothiol glutaredoxins, Grx6 and Grx7, showed a high glutaredoxin activity in vitro. Grx6 and Grx7 overlap in their activity and deletion mutants lacking both proteins show growth defects and a strongly increased sensitivity toward oxidizing agents such as hydrogen peroxide or diamide. Our observations suggest that Grx6 and Grx7 do not play a general role in the oxidative folding of proteins in the early secretory pathway but rather counteract the oxidation of specific thiol groups in substrate proteins. PMID- 18400947 TI - Biomechanical performance of rotator cuff repairs with humeral rotation: a new rotator cuff repair failure model. AB - BACKGROUND: Traditional biomechanical evaluations of rotator cuff repair techniques employ cyclic loading of the supraspinatus tendon in an isolated medial direction. PURPOSE: This study was conducted to evaluate 2 different rotator cuff repair techniques that are currently the subject of debate with cyclic loading and with internal and external humeral rotation to better simulate postoperative rehabilitation. STUDY DESIGN: Controlled laboratory study. METHODS: Nine fresh-frozen paired human cadaver shoulders (18 shoulders) were studied. A single-row repair with 2 suture anchors was compared with a double-row repair with 4 suture anchors. The shoulders were tested in a custom device to position the shoulder in neutral, 45 degrees of internal rotation, and 45 degrees of external rotation. Cyclic loading of the supraspinatus tendon was performed with an MTS material testing machine. Gap formation was measured and analyzed for each rotational position using the MTS device. RESULTS: For the single-row repair, average gap formation per 100 cycles in the positions of neutral, internal, and external humeral rotation was 1.47 +/- 0.63, 3.11 +/- 1.55, and 2.24 +/- 0.94 mm, respectively. For the double-row repair, average gap formation per 100 cycles in the positions of neutral, internal, and external humeral rotation was 1.25 +/- 0.54, 2.29 +/- 1.10, and 1.57 +/- 0.48 mm, respectively. For gapping averaged over all positions, the double-row repair had significantly less gapping than the single-row repair (P = .0109); gapping was greatest for internal rotation, followed by external rotation, and least for neutral (P < .0001). CONCLUSION: The testing method of including a rotational component in biomechanical rotator cuff repair testing is a more realistic model of the loading conditions experienced by a repaired rotator cuff as the patient participates in postoperative rehabilitation. Double-row repair has better fixation strength than single-row repairs when exposed to cyclic loading and changes in humeral rotation position. CLINICAL RELEVANCE: Humeral rotation affects rotator cuff fixation and should be considered in postoperative rehabilitation. PMID- 18400946 TI - ERdj4 and ERdj5 are required for endoplasmic reticulum-associated protein degradation of misfolded surfactant protein C. AB - Mutations in the SFTPC gene associated with interstitial lung disease in human patients result in misfolding, endoplasmic reticulum (ER) retention, and degradation of the encoded surfactant protein C (SP-C) proprotein. In this study, genes specifically induced in response to transient expression of two disease associated mutations were identified by microarray analyses. Immunoglobulin heavy chain binding protein (BiP) and two heat shock protein 40 family members, endoplasmic reticulum-localized DnaJ homologues ERdj4 and ERdj5, were significantly elevated and exhibited prolonged and specific association with the misfolded proprotein; in contrast, ERdj3 interacted with BiP, but it did not associate with either wild-type or mutant SP-C. Misfolded SP-C, ERdj4, and ERdj5 coprecipitated with p97/VCP indicating that the cochaperones remain associated with the misfolded proprotein until it is dislocated to the cytosol. Knockdown of ERdj4 and ERdj5 expression increased ER retention and inhibited degradation of misfolded SP-C, but it had little effect on the wild-type protein. Transient expression of ERdj4 and ERdj5 in X-box binding protein 1(-/-) mouse embryonic fibroblasts substantially restored rapid degradation of mutant SP-C proprotein, whereas transfection of HPD mutants failed to rescue SP-C endoplasmic reticulum associated protein degradation. ERdj4 and ERdj5 promote turnover of misfolded SP C and this activity is dependent on their ability to stimulate BiP ATPase activity. PMID- 18400948 TI - The effect of donor age and low-dose gamma irradiation on the initial biomechanical properties of human tibialis tendon allografts. AB - BACKGROUND: Most tissue banks recover and irradiate tibialis tendon allografts from donors aged up to 65 years. It is unknown whether donor age and low-dose gamma irradiation affect the initial biomechanical properties of tibialis allografts. HYPOTHESIS: Donor age up to 65 years and low-dose gamma irradiation do not significantly affect the initial biomechanical properties of tibialis allografts. STUDY DESIGN: Controlled laboratory study. METHODS: One hundred twenty-six tibialis tendon allografts (63 pairs, 37 human donors) were divided into 3 age groups: young (<45 years), middle (46-55 years), and old (56-65 years). Within each age group, half of the paired tendons underwent tensile testing as single-strand grafts and the other half as double-strand grafts. One tendon from each donor pair was randomly assigned to undergo terminal sterilization with an absorbed dose of 1.46 to 1.80 Mrad (14.6-18.0 kGy) gamma irradiation, whereas the other tendon received no irradiation. All tendon grafts were preconditioned with a cyclic load and tested to failure in tension. RESULTS: Irradiated single-strand tendons in the old age group had a longer displacement at failure compared to the middle but not the young age group. Nonirradiated double-strand tendons in the old age group had a lower failure stress. Single strand irradiated old tendons had a lower stiffness, and all irradiated young tendons and old double-strand tendons had a higher failure stress compared to nonirradiated tendons. CONCLUSION: Donor age up to 65 years does not significantly affect the initial failure load, stiffness, or displacement at failure of tibialis allografts. An age-related decrease in failure stress was observed among nonirradiated tendons but not in tendons subjected to irradiation. CLINICAL RELEVANCE: The results provide biomechanical evidence for use of tibialis allografts from donors up to 65 years of age. Low-dose gamma irradiation does not negatively influence the initial biomechanical properties of tibialis allografts. Further studies examining age and irradiation effects after submaximal cyclic loading conditions are recommended. PMID- 18400949 TI - The effect of anterior cruciate ligament deficiency and reconstruction on the patellofemoral joint. AB - BACKGROUND: Little is known about the effect of anterior cruciate ligament deficiency and reconstruction on the patellofemoral joint. HYPOTHESIS: Anterior cruciate ligament deficiency changes the patellofemoral joint biomechanics. Reconstruction of the ligament does not restore the altered patellofemoral joint function. STUDY DESIGN: Controlled laboratory study. METHODS: Eight patients with an acute anterior cruciate ligament injury in 1 knee and the contralateral side intact were included in the study. Magnetic resonance and dual-orthogonal fluoroscopic imaging techniques were used to compare the patellofemoral joint function during a single-leg lunge between the intact, the anterior cruciate ligament-injured, and the anterior cruciate ligament-reconstructed knee. Data on the patellar tendon apparent elongation and orientation, patellar tracking, and patellofemoral cartilage contact location were collected preoperatively and at 6 months after reconstruction. RESULTS: Anterior cruciate ligament deficiency caused a significant apparent elongation and change in orientation of the patellar tendon. It decreased the flexion and increased the valgus rotation and tilt of the patella. Anterior cruciate ligament injury caused a proximal and lateral shift in patellofemoral cartilage contact location. Anterior cruciate ligament reconstruction reduced the abnormal apparent elongation but not the orientation of the patellar tendon, and it restored the patellar flexion and proximal shift in contact. The abnormal patellar rotation, tilt, and lateral shift in cartilage contact persisted after reconstruction. CONCLUSION: The altered function of the patellar tendon in anterior cruciate ligament deficiency resulted in an altered patellar tracking and patellofemoral cartilage contact. Persistent changes in patellofemoral joint function after anterior cruciate ligament reconstruction imply that reconstruction of the anterior cruciate ligament does not restore the normal function of the patellofemoral joint. CLINICAL RELEVANCE: The abnormal kinematics of the patellofemoral joint might predispose the patellofemoral cartilage to degenerative changes associated with anterior cruciate ligament deficiency, even if the ligament is reconstructed in a way that restores anteroposterior knee laxity. PMID- 18400950 TI - Diffusion of the second messengers in the cytoplasm acts as a variability suppressor of the single photon response in vertebrate phototransduction. AB - The single photon response in vertebrate phototransduction is highly reproducible despite a number of random components of the activation cascade, including the random activation site, the random walk of an activated receptor, and its quenching in a random number of steps. Here we use a previously generated and tested spatiotemporal mathematical and computational model to identify possible mechanisms of variability reduction. The model permits one to separate the process into modules, and to analyze their impact separately. We show that the activation cascade is responsible for generation of variability, whereas diffusion of the second messengers is responsible for its suppression. Randomness of the activation site contributes at early times to the coefficient of variation of the photoresponse, whereas the Brownian path of a photoisomerized rhodopsin (Rh*) has a negligible effect. The major driver of variability is the turnoff mechanism of Rh*, which occurs essentially within the first 2-4 phosphorylated states of Rh*. Theoretically increasing the number of steps to quenching does not significantly decrease the corresponding coefficient of variation of the effector, in agreement with the biochemical limitations on the phosphorylated states of the receptor. Diffusion of the second messengers in the cytosol acts as a suppressor of the variability generated by the activation cascade. Calcium feedback has a negligible regulatory effect on the photocurrent variability. A comparative variability analysis has been conducted for the phototransduction in mouse and salamander, including a study of the effects of their anatomical differences such as incisures and photoreceptors geometry on variability generation and suppression. PMID- 18400951 TI - Binding cooperativity in phage lambda is not sufficient to produce an effective switch. AB - In the wild-type phage lambda, binding of CI to O(R)2 helps polymerase bound to P(RM) transition from a closed to open complex. Activators on other promoters increase the polymerase-DNA binding energy, or affect both the binding energy and the closed-open transition probability. Using a validated mathematical model, we show that these two modes of upregulation have very different effects on the promoter function. We predict that if CI(2) bound to O(R)2 produced equal increase in RNAP-DNA binding constant (compared to wild-type increase in the closed-open transition probability), the lysogen would be significantly less stable. PMID- 18400952 TI - A residue at the cytoplasmic entrance of BK-type channels regulating single channel opening by its hydrophobicity. AB - Single large-conductance calcium-activated K(+) (BK) channels encoded by the mSlo gene usually have synchronous gating, but a Drosophila dSlo (A2/C2/E2/G5/10) splice variant (dSlo1A) exhibits very flickery openings. To probe this difference in gating, we constructed a mutant I323T. This channel exhibits four subconductance levels similar to those of dSlo1A. Rectification of the single channel current-voltage relation of I323T decreased as [Ca(2+) ](in) increased from 10 to 300 microM. Mutagenesis suggests that the hydrophobicity of the residue at the position is important for the wild-type gating; i.e., increasing hydrophobicity prolongs open duration. Molecular dynamics simulation suggests that four hydrophobic pore-lining residues at position 323 of mSlo act cooperatively in a "shutter-like" mechanism gating the permeation of K(+) ions. Rate-equilibrium free energy relations analysis shows that the four I323 residues in an mSlo channel have a conformation 65% similar to the closed conformation during gating. Based on these observations, we suggest that the appearance of rectification and substates of BK-type channels arise from a reduction of the cooperativity among these four residues and a lower probability of being open. PMID- 18400953 TI - Sensation-targeted motor control: every spike counts? Focus on: "whisker movements evoked by stimulation of single motor neurons in the facial nucleus of the rat". PMID- 18400954 TI - Two kinds of FMRI repetition suppression? Evidence for dissociable neural mechanisms. AB - Repetition suppression (RS) is a reduction of neural response that is often observed when stimuli are presented more than once. Many functional magnetic resonance imaging (fMRI) studies have exploited RS to probe the sensitivity of cortical regions to variations in different stimulus dimensions; however, the neural mechanisms underlying fMRI-RS are not fully understood. Here we test the hypothesis that long-interval (between-trial) and short-interval (within-trial) RS effects are caused by distinct and independent neural mechanisms. Subjects were scanned while viewing visual scenes that were repeated over both long and short intervals. Within the parahippocampal place area (PPA) and other brain regions, suppression effects relating to both long- and short-interval repetition were observed. Critically, two sources of evidence indicated that these effects were engendered by different underlying mechanisms. First, long- and short interval RS effects were entirely noninteractive even although they were measured within the same set of trials during which subjects performed a constant behavioral task, thus fulfilling the formal requirements for a process dissociation. Second, long- and short-interval RS were differentially sensitive to viewpoint: short-interval RS was only significant when scenes were repeated from the same viewpoint while long-interval RS less viewpoint-dependent. Taken together, these results indicate that long- and short-interval fMRI-RS are mediated by different neural mechanisms that independently modulate the overall fMRI signal. These findings have important implications for understanding the results of studies that use fMRI-RS to explore representational spaces. PMID- 18400956 TI - Reference frame conversions for repeated arm movements. AB - The aim of this study was to further understand how the brain represents spatial information for shaping aiming movements to targets. Both behavioral and neurophysiological studies have shown that the brain represents spatial memory for reaching targets in an eye-fixed frame. To date, these studies have only shown how the brain stores and updates target locations for generating a single arm movement. But once a target's location has been computed relative to the hand to program a pointing movement, is that information reused for subsequent movements to the same location? Or is the remembered target location reconverted from eye to motor coordinates each time a pointing movement is made? To test between these two possibilities, we had subjects point twice to the remembered location of a previously foveated target after shifting their gaze to the opposite side of the target site before each pointing movement. When we compared the direction of pointing errors for the second movement to those of the first, we found that errors for each movement varied as a function of current gaze so that pointing endpoints fell on opposite sides of the remembered target site in the same trial. Our results suggest that when shaping multiple pointing movements to the same location the brain does not use information from the previous arm movement such as an arm-fixed representation of the target but instead mainly uses the updated eye-fixed representation of the target to recalculate its location into the appropriate motor frame. PMID- 18400955 TI - NMDA receptor desensitization regulated by direct binding to PDZ1-2 domains of PSD-95. AB - Regulation of N-methyl-d-aspartate receptor (NMDAR) activity by desensitization is important in physiological and pathological states; NMDAR desensitization contributes in shaping synaptic responses and may be protective by limiting calcium influx during sustained glutamate insults. We previously reported that glycine-independent desensitization decreases during hippocampal neuronal development, correlating with NMDAR synaptic localization and association with postsynaptic density 95 (PSD-95). PSD-95/Discs large/zona occludens (PDZ)-1,2 domains of PSD-95 bind to the C-terminus of NMDAR NR2 subunits. The role of PSD 95 in anchoring signaling proteins near NMDARs is well documented. To determine if PSD-95-induced changes in NMDAR desensitization occur because of direct binding to NR2 or due to recruitment of regulatory proteins, we tested the effects of various PSD-95 constructs on NMDAR currents in human embryonic kidney 293 (HEK293) cells and neurons. In HEK cells, wild-type PSD-95 significantly reduced wild-type NMDAR desensitization without altering currents of NMDARs containing NR2A-S1462A, a mutation that abolishes PSD-95 binding. The PSD-95 N terminus truncated after the PDZ1-2 domains was sufficient for this effect in neurons with low endogenous PSD-95 levels; in NMDAR-expressing HEK cells, the effect persisted when PSD-95 multimerization was eliminated. Moreover other PSD 95 family members with highly homologous PDZ1-2 domains significantly reduced NMDAR desensitization. In mature neurons, disruption of PSD-95/NMDAR interaction through protein kinase C (PKC) activation increased desensitization to levels found in immature neurons, and this effect was not due to PKC direct regulation of NMDAR activity. We conclude that direct binding of PSD-95 increases stability of NMDAR responses to agonist exposure in neuronal and nonneuronal cells. PMID- 18400957 TI - Developmental changes in the fidelity and short-term plasticity of GABAergic synapses in the neonatal rat dorsal horn. AB - The lower thresholds and increased excitability of dorsal horn neurons in the neonatal rat suggest that inhibitory processing is less efficient in the immature spinal cord. This is unlikely to be explained by an absence of functional GABAergic inhibition because antagonism of gamma-aminobutyric acid (GABA) type A receptors augments neuronal firing in vivo from the first days of life. However, it is possible that more subtle deficits in GABAergic signaling exist in the neonate, such as decreased reliability of transmission or greater depression during repetitive stimulation, both of which could influence the relative excitability of the immature spinal cord. To address this issue we examined monosynaptic GABAergic inputs onto superficial dorsal horn neurons using whole cell patch-clamp recordings made in spinal cord slices at a range of postnatal ages (P3, P10, and P21). The amplitudes of evoked inhibitory postsynaptic currents (IPSCs) were significantly lower and showed greater variability in younger animals, suggesting a lower fidelity of GABAergic signaling at early postnatal ages. Paired-pulse ratios were similar throughout the postnatal period, whereas trains of stimuli (1, 5, 10, and 20 Hz) revealed frequency-dependent short-term depression (STD) of IPSCs at all ages. Although the magnitude of STD did not differ between ages, the recovery from depression was significantly slower at immature GABAergic synapses. These properties may affect the integration of synaptic inputs within developing superficial dorsal horn neurons and thus contribute to their larger receptive fields and enhanced afterdischarge. PMID- 18400958 TI - Dopamine increases the gain of the input-output response of rat prefrontal pyramidal neurons. AB - Dopaminergic modulation of prefrontal cortical activity is known to affect cognitive functions like working memory. Little consensus on the role of dopamine modulation has been achieved, however, in part because quantities directly relating to the neuronal substrate of working memory are difficult to measure. Here we show that dopamine increases the gain of the frequency-current relationship of layer 5 pyramidal neurons in vitro in response to noisy input currents. The gain increase could be attributed to a reduction of the slow afterhyperpolarization by dopamine. Dopamine also increases neuronal excitability by shifting the input-output functions to lower inputs. The modulation of these response properties is mainly mediated by D1 receptors. Integrate-and-fire neurons were fitted to the experimentally recorded input-output functions and recurrently connected in a model network. The gain increase induced by dopamine application facilitated and stabilized persistent activity in this network. The results support the hypothesis that catecholamines increase the neuronal gain and suggest that dopamine improves working memory via gain modulation. PMID- 18400960 TI - Serotonin transduction cascades mediate variable changes in pyloric network cycle frequency in response to the same modulatory challenge. AB - A fundamental question in systems biology addresses the issue of how flexibility is built into modulatory networks such that they can produce context-dependent responses. Here we examine flexibility in the serotonin (5-HT) response system that modulates the cycle frequency (cf) of a rhythmic motor output. We found that depending on the preparation, the same 5-min bath application of 5-HT to the pyloric network of the California spiny lobster, Panulirus interruptus, could produce a significant increase, decrease, or no change in steady-state cf relative to baseline. Interestingly, the mean circuit output was not significantly different among preparations prior to 5-HT application. We developed pharmacological tools to examine the preparation-to-preparation variability in the components of the 5-HT response system. We found that the 5-HT response system consisted of at least three separable components: a 5 HT(2betaPan)-like component mediated a rapid decrease followed by a sustained increase in cf; a 5-HT(1alphaPan)-like component produced a small and usually gradual increase in cf; at least one other component associated with an unknown receptor mediated a sustained decrease in cf. The magnitude of the change in cf produced by each component was highly variable, so that when summed they could produce either a net increase, decrease, or no change in cf depending on the preparation. Overall, our research demonstrates that the balance of opposing components of the 5-HT response system determines the direction and magnitude of 5-HT-induced change in steady-state cf relative to baseline. PMID- 18400959 TI - Computational role of large receptive fields in the primary somatosensory cortex. AB - Computational studies are challenging the intuitive view that neurons with broad tuning curves are necessarily less discriminative than neurons with sharp tuning curves. In the context of somatosensory processing, broad tuning curves are equivalent to large receptive fields. To clarify the computational role of large receptive fields for cortical processing of somatosensory information, we recorded ensembles of single neurons from the infragranular forelimb/forepaw region of the rat primary somatosensory cortex while tactile stimuli were separately delivered to different locations on the forelimbs/forepaws under light anesthesia. We specifically adopted the perspective of individual columns/segregates receiving inputs from multiple body location. Using single trial analyses of many single-neuron responses, we obtained two main results. 1) The responses of even small populations of neurons recorded from within the same estimated column/segregate can be used to discriminate between stimuli delivered to different surround locations in the excitatory receptive fields. 2) The temporal precision of surround responses is sufficiently high for spike timing to add information over spike count in the discrimination between surround locations. This surround spike-timing code (i) is particularly informative when spike count is ambiguous, e.g., in the discrimination between close locations or when receptive fields are large, (ii) becomes progressively more informative as the number of neurons increases, (iii) is a first-spike code, and (iv) is not limited by the assumption that the time of stimulus onset is known. These results suggest that even though large receptive fields result in a loss of spatial selectivity of single neurons, they can provide as a counterpart a sophisticated temporal code based on latency differences in large populations of neurons without necessarily sacrificing basic information about stimulus location. PMID- 18400961 TI - Postnatal changes in the inactivation properties of voltage-gated sodium channels contribute to the mature firing pattern of spinal motoneurons. AB - Most mammals are born with the necessary spinal circuitry to produce a locomotor like pattern of neural activity. However, rodents seldom demonstrate weight supported locomotor behavior until the second or third postnatal week, possibly due to the inability of the neuromuscular system to produce sufficient force during this early postnatal period. As spinal motoneurons mature they are seen to fire an increasing number of action potentials at an increasing rate, which is a necessary component of greater force production. The mechanisms responsible for this enhanced ability of motoneurons are not completely defined. In the present study we assessed the biophysical properties of the developing voltage-gated sodium current to determine their role in the maturing firing pattern. Using dissociated postnatal lumbar motoneurons in short-term culture (18-24 h) we demonstrate that currents recorded from the most mature postnatal age group (P10 P12) were significantly better able to maintain channels in an available state during repetitive stimulation than were the younger age groups (P1-P3, P4-P6, P7 P9). This ability correlated with the ability of channels to recover more quickly and more completely from an inactivated state. These age-related differences were seen in the absence of changes in the voltage dependence of channel gating. Differences in both closed-state inactivation and slow inactivation were also noted between the age groups. The results indicate that changes in the inactivation properties of voltage-gated sodium channels are important for the development of a mature firing pattern in spinal motoneurons. PMID- 18400962 TI - Models and measurements of functional maps in V1. AB - The organization of primary visual cortex has been heavily studied for nearly 50 years, and in the last 20 years functional imaging has provided high-resolution maps of its tangential organization. Recently, however, the usefulness of maps like those of orientation and spatial frequency (SF) preference has been called into question because they do not, by themselves, predict how moving images are represented in V1. In this review, we discuss a model for cortical responses (the spatiotemporal filtering model) that specifies the types of cortical maps needed to predict distributed activity within V1. We then review the structure and interrelationships of several of these maps, including those of orientation, SF, and temporal frequency preference. Finally, we discuss tests of the model and the sufficiency of the requisite maps in predicting distributed cortical responses. Although the spatiotemporal filtering model does not account for all responses within V1, it does, with reasonable accuracy, predict population responses to a variety of complex stimuli. PMID- 18400963 TI - Studies of Lewis antigens and H. pylori adhesion in CHO cell lines engineered to express Lewis b determinants. AB - Many microbes bind and adhere via adhesins to host cell carbohydrates as an initial step for infection. Therefore, cell lines expressing Lewis b (Le(b)) determinants were generated as a potential model system for Helicobacter pylori colonization and infection, and their expression of blood group Lewis determinants was characterized. CHO-K1 cells were stably transfected with selected glycosyltransferase cDNAs, and two Le(b) positive clones, 1C5 and 2C2, were identified. Expression of Lewis (Le(a), Le(b), Le(x), and Le(y)) determinants was analyzed by flow cytometry of intact cells, SDS-PAGE/Western blot of solubilized glycoproteins, and thin layer chromatography immunostaining of isolated glycolipids (GL). Binding of H. pylori to cells was examined by microscopy and quantified. Flow cytometry showed that 1C5 and 2C2 were Le(a) and Le(b) positive. 1C5 expressed Le(b) on O-linked, but not N-linked, glycans and only weakly on GLs. In contrast, 2C2 expressed Le(b) on N-, O-glycans, and GLs. Furthermore, both clones expressed Le(a) on N- and O-glycans but not on GLs. 2C2, but not 1C5, stained positively for Le(y) on N-linked glycans and GLs. Both clones, as well as the parental CHO-K1 cells, expressed Le(x) on GLs. A Le(b) binding H. pylori strain bound to the 1C5 and 2C2 cells. In summary, two glycosyltransferase transfected CHO-K1 cell clones differed regarding Lewis antigen expression on N- and O-linked glycans as well as on GLs. Both clones examined supported adhesion of a Le(b)-binding H. pylori strain and may thus be a useful in vitro model system for H. pylori colonization/infection studies. PMID- 18400964 TI - A randomized study of extended dosing regimens for initiation of epoetin alfa treatment for anemia of chronic kidney disease. AB - BACKGROUND AND OBJECTIVES: Although epoetin alfa is commonly initiated weekly (QW) in anemic chronic kidney disease (CKD) patients, recent evidence indicates that it can be initiated every 2 wk (Q2W) and used in maintenance therapy every 4 wk (Q4W). This study examined the feasibility of initiating epoetin alfa Q4W in anemic CKD patients not receiving dialysis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This open-label study randomized subjects (1:2:2:2) to treatment with epoetin alfa 10,000 IU QW, 20,000 IU Q2W, 20,000 IU Q4W, or 40,000 IU Q4W for 16 wk. Subjects were > or =18 yr, had hemoglobin <11 g/dl, a glomerular filtration rate of 15 to 90 ml/min per 1.73 m(2), and had not received erythropoietic therapy within 8 wk. The primary analysis was a noninferiority comparison of the 40,000 IU Q4W to the 20,000 IU Q2W group in the per-protocol population with respect to hemoglobin change from baseline to the end of study. RESULTS: Of 262 subjects randomized, 229 comprised the per-protocol population. Mean hemoglobin change from baseline for the 40,000 IU Q4W group (1.24 g/dl) was not inferior to the 20,000 IU Q2W group (1.11 g/dl) with the lower limit of 95% CI, -0.21 g/dl. In the QW, 20,000 IU Q2W, 20,000 IU Q4W, and 40,000 IU Q4W groups, 90%, 87%, 75%, and 86% of subjects, respectively, achieved a hemoglobin increase > or =1 g/dl. Serious adverse events were similar across all groups. CONCLUSIONS: Epoetin alfa can be initiated Q4W in anemic CKD subjects. PMID- 18400965 TI - Comparison of early versus late use of antibiotic locks in the treatment of catheter-related bacteremia. AB - BACKGROUND AND OBJECTIVES: This retrospective study compared the effectiveness of the timing of the antibiotic locks to clear catheter-related bacteremia in children on chronic hemodialysis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The early antibiotic lock group received antibiotic locks along with systemic antibiotics from the very beginning of catheter-related bacteremia. The late antibiotic lock group was given only systemic antibiotics initially, and antibiotic locks were used late in the infection if the catheter-related bacteremia could not be cleared after resolution of symptoms. RESULTS: There were 264 catheter-related bacteremias in 79 children during 6 yr of observation. Early antibiotic locks were able to clear catheter-related bacteremia and resolve the symptoms more effectively without the need for catheter exchange when compared with late antibiotic locks. A total of 84 catheter-related bacteremias required wire-guided exchange of the catheters. Late antibiotic locks required wire-guided catheter exchange more frequently than the early antibiotic locks. The post catheter-related bacteremia infection-free survival of the catheters after wire guided exchange were significantly longer than those of both antibiotic lock groups. Recurrence of catheter-related bacteremia within 45 d after wire-guided exchange occurred at similar rates compared with the antibiotic lock groups. CONCLUSION: Antibiotic locks are significantly more effective in clearing catheter-related bacteremia when used early in infection, diminishing the need for catheter exchange. Wire-guided exchange has a late-onset advantage for infection-free survival compared with catheter in situ treatment. The recurrence rates in the first 45 d after catheter-related bacteremia are similar regardless of the treatment strategy. PMID- 18400966 TI - Albumin excretion rate in normal adolescents: relation to insulin resistance and cardiovascular risk factors and comparisons to type 1 diabetes mellitus patients. AB - BACKGROUND AND OBJECTIVES: Although albumin excretion rates have been related to cardiovascular morbidity and mortality in both diabetic and nondiabetic adults, little is known about the relation between albuminuria and either cardiovascular risk factors or the insulin resistance syndrome in adolescents. A normal range for albumin excretion in adolescents was established, correlations between albumin excretion and cardiovascular risk factors were evaluated, and albumin excretion in normal adolescents was compared with that in type 1 diabetes mellitus adolescents. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Albumin excretion rate was measured in 368 normal and 175 diabetic adolescents. Multiple regression analysis was used to predict the relation of age, sex, Tanner stage, body mass index, and systolic blood pressure to albumin excretion in both cohorts. In addition, correlations between albumin excretion and age, blood pressure, body mass index, lipids, and measurements of insulin resistance were performed in the normal adolescents. RESULTS: Mean albumin excretion was significantly lower in normal adolescents (4.0 microg/min) than in type 1 diabetic adolescents (5.0 microg/min). Albumin excretion increased with age in diabetics. Albumin excretion did not significantly correlate with any measure of cardiovascular risk or insulin resistance but did significantly correlate with fasting insulin. CONCLUSIONS: Albumin excretion rate is not related to insulin resistance or traditional cardiovascular risk factors in adolescence but is related to fasting insulin. Diabetic adolescents have increased albumin excretion compared with normal adolescents. PMID- 18400968 TI - Introduction to Vitamin D Symposium, March 14, 2008. AB - A large body of work in diverse clinical and scientific areas has accumulated that supports a role for vitamin D in multiple organ systems and physiologic and molecular processes. The vitamin D receptor is distributed ubiquitously, and by binding with its receptor, vitamin D initiates a series of events that can affect cellular proliferation and differentiation, inflammation, the immune system, and the endocrine system, including the renin-angiotensin system, insulin resistance, and lipid metabolism. PMID- 18400967 TI - Need for quality improvement in renal systematic reviews. AB - BACKGROUND AND OBJECTIVES: Systematic reviews of clinical studies aim to compile best available evidence for various diagnosis and treatment options. This study assessed the methodologic quality of all systematic reviews relevant to the practice of nephrology published in 2005. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We searched electronic databases (Medline, Embase, American College of Physicians Journal Club, Cochrane) and hand searched Cochrane renal group records. Clinical practice guidelines, case reports, narrative reviews, and pooled individual patient data meta-analyses were excluded. Methodologic quality was measured using a validated questionnaire (Overview Quality Assessment Questionnaire). For reviews of randomized trials, we also evaluated adherence to recommended reporting guidelines (Quality of Reporting of Meta-Analyses). RESULTS: Ninety renal systematic reviews were published in year 2005, 60 of which focused on therapy. Many systematic reviews (54%) had major methodologic flaws. The most common review flaws were failure to assess the methodologic quality of included primary studies and failure to minimize bias in study inclusion. Only 2% of reviews of randomized trials fully adhered to reporting guidelines. A minority of journals (four of 48) endorsed adherence to consensus guidelines for review reporting, and these journals published systematic reviews of higher methodologic quality (P < 0.001). CONCLUSIONS: The majority of systematic reviews had major methodologic flaws. The majority of journals do not endorse consensus guidelines for review reporting in their instructions to authors; however, journals that recommended such adherence published systemic reviews of higher methodologic quality. PMID- 18400969 TI - Association of strong immune responses to PPE protein Rv1168c with active tuberculosis. AB - Accurate diagnosis of tuberculosis (TB) infection is critical for the treatment, prevention, and control of TB. Conventional diagnostic tests based on purified protein derivative (PPD) do not achieve the required diagnostic sensitivity. Therefore, in this study, we have evaluated the immunogenic properties of Rv1168c, a member of the PPE family, in comparison with PPD, which is routinely used in the tuberculin test, and Hsp60 and ESAT-6, well-known immunodominant antigens of Mycobacterium tuberculosis. In a conventional enzyme immunoassay, the recombinant Rv1168c protein displayed stronger immunoreactivity against the sera obtained from patients with clinically active TB than did PPD, Hsp60, or ESAT-6 and could distinguish TB patients from Mycobacterium bovis BCG-vaccinated controls. Interestingly, Rv1168c antigen permits diagnosis of smear-negative pulmonary TB as well as extrapulmonary TB cases, which are often difficult to diagnose by conventional tests. The immunodominant nature of Rv1168c makes it a promising candidate to use in serodiagnosis of TB. In addition, our studies also show that Rv1168c is a potent T-cell antigen which elicits a strong gamma interferon response in sensitized peripheral blood mononuclear cells obtained from TB patients. PMID- 18400971 TI - Enhancement of diagnostic efficiency by a gamma interferon release assay for pulmonary tuberculosis. AB - This study was designed to examine the use of the QuantiFERON-TB Gold assay as an aid in the diagnosis of active pulmonary tuberculosis (TB) in Brazilian patients. Using the receiver operating characteristic curve, the cutoff was adjusted to >or=0.20 IU/ml. The sensitivity increased to 86%, with 100% specificity. All TB patients with negative sputum smear microscopy and negative culture results were positive using this test. PMID- 18400970 TI - Molecular structure and expression of anthropic, ovine, and murine forms of complement receptor type 2. PMID- 18400972 TI - LruA and LruB antibodies in sera of humans with leptospiral uveitis. AB - Uveitis can be a serious complication of leptospirosis. Previous studies indicated that the leptospiral lipoproteins LruA and LruB are expressed in the eyes of uveitic horses and that antibodies directed against those proteins show in vitro cross-reactivity with components of equine lens, ciliary body, and/or retina. We now demonstrate that sera from a significant proportion of humans who have leptospiral uveitis also contain antibodies against LruA and LruB. Different categories of nonleptospiral uveitis and autoimmune uveitis were also screened; patients diagnosed with Fuchs uveitis or Behcet's syndrome produced antibodies that cross-reacted with LruA and LruB, suggesting similarities of the autoimmune responses in those diseases with those of leptospiral uveitis. PMID- 18400974 TI - Differential immune responses of red Deer (Cervus elaphus) following experimental challenge with Mycobacterium avium subsp. paratuberculosis. AB - Immune responses of red deer (Cervus elaphus) that presented with different levels of paucibacillary pathology were profiled to detail immune changes during the progression of Johne's disease. Immune responses were monitored using an immunoglobulin G1 (IgG1) antibody enzyme-linked immunosorbent assay (ELISA), a gamma interferon (IFN-gamma) ELISA, and flow cytometry. Animals in the study were divided into outcome groups postmortem according to disease severity. All animals mounted IgG1 antibody and IFN-gamma responses to both the vaccination and experimental challenges. The Mycobacterium avium subsp. paratuberculosis-specific IgG1 antibody responses in the challenged group showed marked differences between infected and severely diseased animals. Slightly higher IFN-gamma responses were seen in infected animals compared with severely diseased animals. No significant changes were seen in the phenotype of lymphocyte populations investigated. Vaccination with killed M. avium subsp. paratuberculosis in mineral oil adjuvant reduced the level of severe disease; however, it obscured immunological differences between the infected and severely diseased groups. This suggests protection is not exclusively mediated via the presence of a type 1 response and, furthermore, the presence of a type 2 response is compatible with protection. These profiles provide information on the different immune processes in Johne's disease progression. PMID- 18400973 TI - Effects of antenatal and postnatal environments on CD4 T-cell responses to Mycobacterium bovis BCG in healthy infants in the Gambia. AB - The Mycobacterium bovis BCG vaccine has a poor record of efficacy in low-income tropical settings. Against this background, we evaluated the immune response of infants to mycobacterial antigens over the 2 years following BCG vaccination at birth by measuring the gamma interferon (IFN-gamma), interleukin-2 (IL-2), and CD154 responses of CD4 T cells. Similar numbers of cells expressed IFN-gamma in infants, 4- to 5-year-old children, and adults, while CD154 was not expressed at comparable levels until the second year of infancy. The IL-2 response remained relatively low in infants, children, and adults but correlated negatively with mother's body mass index and was highest among infants born to Mandinka mothers. Similarly, infants born in the wet season had a stronger CD154 response than those born in the dry season throughout the 2 years of the study. We conclude that the prenatal and perinatal environments have a lasting effect on the response of infants to the BCG vaccine. PMID- 18400975 TI - Dendritic cells differentiated in the presence of a single-stranded viral RNA sequence conserve their ability to activate CD4 T lymphocytes but lose their capacity for Th1 polarization. AB - Monocyte-derived dendritic cells (DCs) differentiate in the presence of Toll-like receptor (TLR) ligands in the course of ongoing infections. A single-stranded RNA (ssRNA) sequence, corresponding to the sequence of the U5 region of human immunodeficiency virus type 1 RNA, was used to mimic viral activation of TLR7 in human DCs. We determined the effector potential of DCs differentiated in the presence of this ssRNA molecule (ssRNA-DCs). ssRNA-DCs phenotypically resembled mature DCs. In contrast, their capacity to allostimulate naive CD4(+) T cells resembled that of conventional immature DCs and could be increased by TLR4 stimulation. Th1 polarization of CD4(+) T cells and production of interleukin 12p70 (IL-12p70) by ssRNA-DCs were selectively abrogated in response to a late TLR4, but not in response to a CD40, maturation signal. Inhibition of p38 mitogen activated protein kinase partially restored IL-12p70 secretion but did not restore Th1 polarization, whereas addition of exogenous IL-12 led to recovery of Th1 polarization. In contrast to lipopolysaccharide, ssRNA induced IL-12p70 production at the very earliest stages of DC differentiation, indicating a particular role for TLR7 in monocyte-derived DCs recently engaged in differentiation. These data demonstrate generation of phenotypically mature DCs with the ability to expand CD4(+) T lymphocytes lacking Th1/2-polarizing capacity. PMID- 18400976 TI - Clinical phase 1 testing of the safety and immunogenicity of an epitope-based DNA vaccine in human immunodeficiency virus type 1-infected subjects receiving highly active antiretroviral therapy. AB - A DNA vaccine encoding sequence-conserved human immunodeficiency virus type 1 (HIV-1)-derived cytotoxic T-lymphocyte (CTL) epitopes from multiple HIV-1 gene products (designated EP HIV-1090) was evaluated in a placebo-controlled, dose escalation phase 1 clinical trial of HIV-1-infected subjects receiving potent combination antiretroviral therapy. Patients received four intramuscular immunizations with EP HIV-1090 over a 4-month period at one of four doses (0.5, 1.0, 2.0, or 4.0 mg) or received a placebo. The vaccine was determined to be safe and well tolerated at all doses tested. CTL responses were measured from cryopreserved peripheral blood mononuclear cells using gamma interferon enzyme linked immunospot assays, with and without in vitro peptide stimulation (IVS). Responses to one or more vaccine epitopes were detected throughout the course of vaccination in 37.5% (12/32) and 47% (15/32) of vaccine recipients measured without and with IVS, respectively, indicating possible vaccine-induced priming of epitope-specific T cells. However, differences in rates of response to HIV-1 epitopes between vaccine and placebo recipients did not achieve statistical significance. The HIV-1 epitope-specific CTL responses measured in the peripheral blood after vaccination were often low level and short-lived, and therefore, alternative immunization schedules, routes of delivery, or vaccine formulations may be required to increase vaccine potency. PMID- 18400977 TI - Efficient testing of large pools of Mycobacterium tuberculosis RD1 peptides and identification of major antigens and immunodominant peptides recognized by human Th1 cells. AB - Comparative genomics has identified several regions of difference (RDs) of Mycobacterium tuberculosis that are deleted or absent in Mycobacterium bovis BCG vaccines. To determine their relevance for diagnostic and vaccine applications, it is imperative that efficient methods are developed to test the encoded proteins for immunological reactivity. In this study, we have used 220 synthetic peptides covering sequences of 12 open reading frames (ORFs) of RD1 and tested them as a single pool (RD1(pool)) with peripheral blood mononuclear cells obtained from pulmonary tuberculosis (TB) patients and M. bovis BCG-vaccinated healthy subjects in Th1 cell assays that measure antigen-induced proliferation and IFN-gamma secretion. The results showed that RD1(pool) induced strong responses in both TB patients and BCG-vaccinated healthy subjects. The subsequent testing of peptide pools of individual ORFs revealed that all ORFs induced positive responses in a portion of donors, but PPE68, CFP10, and ESAT6 induced strong responses in TB patients and PPE68 induced strong responses in BCG vaccinated healthy subjects. In addition, HLA-DR and -DQ typing of donors and HLA DR binding prediction analysis of proteins suggested HLA-promiscuous presentation of PPE68, CFP10, and ESAT6. Further testing of individual peptides showed that a single peptide of PPE68 (121-VLTATNFFGINTIPIALTEMDYFIR-145) was immunodominant. The search for sequence homology revealed that a part of this peptide, 124 ATNFFGINTIPIAL-137, was present in several PPE family proteins of M. tuberculosis and M. bovis BCG vaccines. Further experiments limited the promiscuous and immunodominant epitope region to the 10-amino-acid cross-reactive sequence 127 FFGINTIPIA-136. PMID- 18400978 TI - Association of mannose-binding lectin gene polymorphism but not of mannose binding serine protease 2 with chronic severe aortic regurgitation of rheumatic etiology. AB - N-Acetylglucosamine (GlcNAc) is the major immunoepitope of group A streptococcal cell wall carbohydrates. Antistreptococcal antibodies cross-reactive with anti GlcNAc and laminin are present in sera of patients with rheumatic fever. The cross-reactivity of these antibodies with human heart valvular endothelium and the underlying basement membrane has been suggested to be a possible cause of immune-mediated valve lesion. Mannose-binding lectin (MBL) encoded by the MBL2 gene, a soluble pathogen recognition receptor, has high affinity for GlcNAc. We postulated that mutations in exon 1 of the MBL2 gene associated with a deficient serum level of MBL may contribute to chronic severe aortic regurgitation (AR) of rheumatic etiology. We studied 90 patients with severe chronic AR of rheumatic etiology and 281 healthy controls (HC) for the variants of the MBL2 gene at codons 52, 54, and 57 by using a PCR-restriction fragment length polymorphism based method. We observed a significant difference in the prevalence of defective MBL2 alleles between patients with chronic severe AR and HC. Sixteen percent of patients with chronic severe AR were homozygotes or compound heterozygotes for defective MBL alleles in contrast to 5% for HC (P = 0.0022; odds ratio, 3.5 [95% confidence interval, 1.6 to 7.7]). No association was detected with the variant of the MASP2 gene. Our study suggests that MBL deficiency may contribute to the development of chronic severe AR of rheumatic etiology. PMID- 18400979 TI - Prevention of alcohol problems in Dutch youth: missed opportunities and new developments. AB - Binge drinking among young people is a problem in the Netherlands. This article outlines the current Dutch approach to alcohol prevention in this target group. It is argued that well-enforced evidence-based control measures are lacking despite renewed political interest in them. Politicians often favor alcohol education, but to increase the effectiveness of alcohol prevention, a combined approach of policy measures, enforcement, and education is needed. Translation of education and policy-based measures is discussed. PMID- 18400982 TI - Effects of once-daily oral and transdermal methylphenidate on sleep behavior of children with ADHD. AB - OBJECTIVE: Methylphenidate is a leading first-line treatment for ADHD (AD/HD). This stimulant has long been suspected to adversely affect sleeping patterns of treated individuals, especially children. There are few studies on the effects of recently developed longer-acting methylphenidate treatments, such as once-daily oral or transdermal formulations, on sleep. METHOD: The authors examined eight indices of sleep behavior among children treated with either of these two methylphenidate preparations or placebo in a randomized, double-blind, multicenter, parallel-group study. RESULTS: The main predictor of sleep problems was baseline numbers or severity of preexisting sleep problems, whereas the different treatments and placebo varied little in their propensity to elicit such problems. There was no significant relationship between dosage and severity or frequency of sleep problems. CONCLUSION: The authors found little evidence that methylphenidate treatment (at least in sustained-release forms) was a significant cause of sleep problems in treated children who were carefully titrated to an optimal dose. PMID- 18400983 TI - Vigilance and sustained attention in children and adults with ADHD. AB - OBJECTIVE: The present article tests the hypothesis of a sustained attention deficit in children and adults suffering from ADHD. METHOD: Vigilance and sustained attention of 52 children with ADHD and 38 adults with ADHD were assessed using a computerized vigilance task. Furthermore, the attentional performance of healthy children (N = 52) and healthy adults (N = 38) was examined. RESULTS: Children and adults with ADHD performed significantly less well in the vigilance task than healthy participants (main effect for group). Furthermore, children and adults showed a significant decrease of performance over time (time-on-task effects). However, there was no greater decrement of performance with the passage of time in patient groups than in control groups (group-by-time interaction). CONCLUSION: The present results do not support the hypothesis of a sustained attention deficit in children and adults with ADHD. PMID- 18400984 TI - Specific signals involved in the long-term maintenance of radiation-induced fibrogenic differentiation: a role for CCN2 and low concentration of TGF-beta1. AB - The fibrogenic differentiation of resident mesenchymal cells is a key parameter in the pathogenesis of radiation fibrosis and is triggered by the profibrotic growth factors transforming growth factor (TGF)-beta1 and CCN2. TGF-beta1 is considered the primary inducer of fibrogenic differentiation and is thought to control its long-term maintenance, whereas CCN2 is considered secondary effector of TGF-beta1. Yet, in long-term established fibrosis like that associated with delayed radiation enteropathy, in situ TGF-beta1 deposition is low, whereas CCN2 expression is high. To explore this apparent paradox, cell response to increasing doses of TGF-beta1 was investigated in cells modeling initiation and maintenance of fibrosis, i.e., normal and fibrosis-derived smooth muscle cells, respectively. Activation of cell-specific signaling pathways by low TGF-beta1 doses was demonstrated with a main activation of the Rho/ROCK pathway in fibrosis-derived cells, whereas the Smad pathway was mainly activated in normal cells. This leads to subsequent and cell-specific regulation of the CCN2 gene. These results suggested a specific profibrotic role of CCN2 in fibrosis-initiated cells. Furthermore, the modulation of CCN2 expression by itself and the combination of TGF-beta1 and CCN2 was investigated in fibrosis-derived cells. In fibrosis initiated cells CCN2 triggered its autoinduction; furthermore, low concentration of TGF-beta1-potentiated CCN2 autoinduction. Our findings showed a differential requirement and action of TGF-beta1 in the fibrogenic response of normal vs. fibrosis-derived cells. This study defines a novel Rho/ROCK but Smad3-independent mode of TGF-beta signaling that may operate during the chronic stages of fibrosis and provides evidence of both specific and combinatorial roles of low TGF-beta1 dose and CCN2. PMID- 18400985 TI - Bestrophin Cl- channels are highly permeable to HCO3-. AB - Bestrophin-1 (Best1) is a Cl(-) channel that is linked to various retinopathies in both humans and dogs. Dysfunction of the Best1 Cl(-) channel has been proposed to cause retinopathy because of altered Cl(-) transport across the retinal pigment epithelium (RPE). In addition to Cl(-), many Cl(-) channels also transport HCO3(-). Because HCO3(-) is physiologically important in pH regulation and in fluid and ion transport across the RPE, we measured the permeability and conductance of bestrophins to HCO3(-) relative to Cl(-). Four human bestrophin homologs (hBest1, hBest2, hBest3, and hBest4) and mouse Best2 (mBest2) were expressed in HEK cells, and the relative HCO3(-) permeability (P HCO3/PCl) and conductance (G HCO3/GCl) were determined. P HCO3/PCl was calculated from the change in reversal potential (Erev) produced by replacing extracellular Cl(-) with HCO3(-). hBest1 was highly permeable to HCO3(-) (P HCO3)/PCl = approximately 0.44). hBest2, hBest4, and mBest2 had an even higher relative HCO3(-) permeability (P HCO3/PCl = 0.6-0.7). All four bestrophins had HCO3(-) conductances that were nearly the same as Cl(-) (G HCO3/GCl = 0.9-1.1). Extracellular Na+ did not affect the permeation of hBest1 to HCO3(-). At physiological HCO3(-) concentration, HCO3(-) was also highly conductive. The hBest1 disease-causing mutations Y85H, R92C, and W93C abolished both Cl(-) and HCO3(-) currents equally. The V78C mutation changed P HCO3/PCl and G HCO3/GCl of mBest2 channels. These results raise the possibility that disease-causing mutations in hBest1 produce disease by altering HCO3(-) homeostasis as well as Cl(-) transport in the retina. PMID- 18400986 TI - Neuronal nitric oxide synthase signaling within cardiac myocytes targets phospholamban. AB - Studies have shown that neuronal nitric oxide synthase (nNOS, NOS1) knockout mice (NOS1-/-) have increased or decreased contractility, but consistently have found a slowed rate of intracellular Ca2+ ([Ca2+]i) decline and relengthening. Contraction and [Ca2+]i decline are determined by many factors, one of which is phospholamban (PLB). The purpose of this study is to determine the involvement of PLB in the NOS1-mediated effects. Force-frequency experiments were performed in trabeculae isolated from NOS1-/- and wild-type (WT) mice. We also simultaneously measured Ca2+ transients (Fluo-4) and cell shortening (edge detection) in myocytes isolated from WT, NOS1-/-, and PLB-/- mice. NOS1-/- trabeculae had a blunted force-frequency response and prolonged relaxation. We observed similar effects in myocytes with NOS1 knockout or specific NOS1 inhibition with S-methyl l-thiocitrulline (SMLT) in WT myocytes (i.e., decreased Ca2+ transient and cell shortening amplitudes and prolonged decline of [Ca2+]i). Alternatively, NOS1 inhibition with SMLT in PLB-/- myocytes had no effect. Acute inhibition of NOS1 with SMLT in WT myocytes also decreased basal PLB serine16 phosphorylation. Furthermore, there was a decreased SR Ca2+ load with NOS1 knockout or inhibition, which is consistent with the negative contractile effects. Perfusion with FeTPPS (peroxynitrite decomposition catalyst) mimicked the effects of NOS1 knockout or inhibition. beta-Adrenergic stimulation restored the slowed [Ca2+]i decline in NOS1-/- myocytes, but a blunted contraction remained, suggesting additional protein target(s). In summary, NOS1 inhibition or knockout leads to decreased contraction and slowed [Ca2+]i decline, and this effect is absent in PLB-/- myocytes. Thus NOS1 signaling modulates PLB serine16 phosphorylation, in part, via peroxynitrite. PMID- 18400987 TI - Induction of Na+/K+/2Cl- cotransporter expression mediates chronic potentiation of intestinal epithelial Cl- secretion by EGF. AB - Alterations in EGF receptor (EGFR) signaling occur in intestinal disorders associated with dysregulated epithelial transport. In the present study, we investigated a role for the EGFR in the chronic regulation of intestinal epithelial secretory function. Epithelial Cl(-) secretion was measured as changes in short-circuit current (Isc) across voltage-clamped monolayers of T84 cells in Ussing chambers. Acute treatment of T84 cells with EGF (100 ng/ml, 15 min) chronically enhanced Isc responses to a broad range of secretagogues. This effect was apparent within 3 h, maximal by 6 h, and sustained for 24 h after treatment with EGF. The Na+/K+/2Cl(-) cotransporter (NKCC1) inhibitor bumetanide (100 microM) abolished the effect of EGF, indicating increased responses are due to potentiated Cl(-) secretion. Neither basal nor agonist-stimulated levels of intracellular Ca2+ or PKA activity were altered by EGF, implying that the effects of the growth factor are not due to chronic alterations in levels of second messengers. EGF increased the expression of NKCC1 with a time course similar to that of its effects on Cl(-) secretion. This effect of EGF was maximal after 6 h, at which time NKCC1 expression in EGF-treated cells was 199.9 +/- 21.9% of that in control cells (n = 21, P < 0.005). EGF-induced NKCC1 expression was abolished by actinomycin D, and RT-PCR analysis demonstrated EGF increased expression of NKCC1 mRNA. These data increase our understanding of mechanisms regulating intestinal fluid and electrolyte transport and reveal a novel role for the EGFR in the chronic regulation of epithelial secretory capacity through upregulation of NKCC1 expression. PMID- 18400988 TI - Maintaining the integrity of trophoblast during growth of the placenta. Focus on "Insulin-like growth factor I and II regulate the life cycle of trophoblast in the developing human placenta". PMID- 18400989 TI - Enhanced exocytotic-like insertion of Orai1 into the plasma membrane upon intracellular Ca2+ store depletion. AB - Ca+ release-activated Ca2+ (CRAC) channels are activated when free Ca2+ concentration in the intracellular stores is substantially reduced and mediate sustained Ca2+ entry. Recent studies have identified Orai1 as a CRAC channel subunit. Here we demonstrate that passive Ca2+ store depletion using the inhibitor of the sarcoendoplasmic reticulum Ca2+-ATPase, thapsigargin (TG), enhances the surface expression of Orai1, a process that depends on rises in cytosolic free Ca2+ concentration, as demonstrated in cells loaded with dimethyl BAPTA, an intracellular Ca2+ chelator that prevented TG-evoked cytosolic free Ca2+ concentration elevation. Similar results were observed with a low concentration of carbachol. Cleavage of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor, synaptosomal-assiciated protein-25 (SNAP-25), with botulinum neurotoxin A impaired TG-induced increase in the surface expression of Orai1. In addition, SNAP-25 cleaving by botulinum neurotoxin A reduces the maintenance but not the initial stages of store-operated Ca2+ entry. In aggregate, these findings demonstrate that store depletion enhances Orai1 plasma membrane expression in an exocytotic manner that involves SNAP-25, a process that contributes to store-dependent Ca2+ entry. PMID- 18400990 TI - Insulin-like growth factor I and II regulate the life cycle of trophoblast in the developing human placenta. AB - The main disorders of human pregnancy are rooted in defective placentation. Normal placental development depends on proliferation, differentiation, and fusion of cytotrophoblasts to form and maintain an overlying syncytiotrophoblast. There is indirect evidence that the insulin-like growth factors (IGFs), which are aberrant in pregnancy disorders, are involved in regulating trophoblast turnover, but the processes that control human placental growth are poorly understood. Using an explant model of human first-trimester placental villus in which the spatial and ontological relationships between cell populations are maintained, we demonstrate that cytotrophoblast proliferation is enhanced by IGF-I/IGF-II and that both factors can rescue cytotrophoblast from apoptosis. Baseline cytotrophoblast proliferation ceases in the absence of syncytiotrophoblast, although denuded cytotrophoblasts can proliferate when exposed to IGF and the rate of cytotrophoblast differentiation/fusion and, consequently, syncytial regeneration, increases. Use of signaling inhibitors suggests that IGFs mediate their effect on cytotrophoblast proliferation/syncytial formation through the MAPK pathway, whereas effects on survival are regulated by the phosphoinositide 3 kinase pathway. These results show that directional contact between cytotrophoblast and syncytium is important in regulating the relative amounts of the two cell populations. However, IGFs can exert an exogenous regulatory influence on placental growth/development, suggesting that manipulation of the placental IGF axis may offer a potential therapeutic route to the correction of inadequate placental growth. PMID- 18400992 TI - Structural and functional changes in regenerating antennules in the crayfish Orconectes sanborni. AB - Crayfish rely on the chemosensory neurons in their antennules to help them find food and habitat and to mediate social interactions. These structures often sustain damage from aggressive interactions or from the environment, but they have the ability to regenerate. In this study, we examine whether the effects of antennule ablation and regeneration on odor-tracking ability correlate with structural changes in the antennule that occur during regeneration. We initiated the regeneration process by removing the right antennules from 55 individuals of Orconectes sanborni. We developed a method to nondestructively sample the regenerating antennules so that we could follow the growth of new antennular tissue in the same animals over time. We used dental epoxy to make molds of the regenerating antennule after each molt. We then made resin positives, which were visualized using scanning electron microscopy. Structural parameters including aesthetasc length, diameter, segment length, and number per row were measured from scanning electron micrographs using Image J software. Crayfish were tested in a tabletop water Y-maze before and after surgery and after each molt to assess their ability to track food odors. The structural and the behavioral data indicate that the antennules possessed many aspects of their original structure by the end of the second molt. Flicking of antennules, investigation of substrate, success rate at finding the odor-containing Y-maze branch, and time to completion of Y-maze regained pre-antennulectomy values by the end of the third molt. PMID- 18400993 TI - A spring-matrix model for pigment translocation in the red ovarian chromatophores of the freshwater shrimp Macrobrachium olfersi (Crustacea, Decapoda). AB - A model for intracellular transport of pigment granules in the red ovarian chromatophores of the freshwater shrimp Macrobrachium olfersi is proposed on the basis of shifts in the equilibrium of resting forces acting on an elastic pigment matrix. The model describes a pigment-transport mechanism in which mechanochemical protein motors like kinesin and myosin alternately stretch and compress a structurally unified, elastic pigment matrix. Quantifiable properties of the spring-matrix obey Hooke's Law during the rapid phases of pigment aggregation and dispersion. The spring-like response of the pigment mass is estimated from previous kinetic experiments on pigment translocation induced by red pigment concentrating hormone, or by the calcium ionophore A23187. Both translocation effectors trigger an initial phase of rapid pigment aggregation, and their removal or washout after complete aggregation produces a phase of rapid pigment dispersion, followed by slow pigment translocation. The rapid-phase kinetics of pigment transport are in reasonable agreement with Hooke's Law, suggesting that such phases represent the release of kinetic energy, probably produced by the mechanochemical protein motors and stored in the form of matrix deformation during the slow phases of translocation. This semiquantitative model should aid in analyzing intracellular transport systems that incorporate an elastic component. PMID- 18400991 TI - Repetitive deformation activates Src-independent FAK-dependent ERK motogenic signals in human Caco-2 intestinal epithelial cells. AB - Repetitive deformation due to villous motility or peristalsis may support the intestinal mucosa, stimulating intestinal epithelial proliferation under normal circumstances and restitution in injured and inflamed mucosa rich in tissue fibronectin. Cyclic strain enhances Caco-2 and IEC-6 intestinal epithelial cell migration across fibronectin via ERK. However, the upstream mediators of ERK activation are unknown. We investigated whether Src and FAK mediate strain induced ERK phosphorylation and migration in human Caco-2 intestinal epithelial cells on fibronectin. Monolayers on tissue fibronectin-precoated membranes were subjected to an average 10% repetitive deformation at 10 cycles/min. Phosphorylation of Src-Tyr 418, FAK-Tyr 397-Tyr 576-Tyr 925, and ERK were significantly increased by deformation. The stimulation of wound closure by strain was prevented by Src blockade with PP2 (10 micromol/l) or specific short interfering (si)RNA. Src inhibition also prevented strain-induced FAK phosphorylation at Tyr 397 and Tyr 576 but not FAK-Tyr 925 or ERK phosphorylation. Reducing FAK by siRNA inhibited strain-induced ERK phosphorylation. Transfection of NH2-terminal tyrosine phosphorylation-deficient FAK mutants Y397F, Y576F-Y577F, and Y397F-Y576F-Y577F did not prevent the activation of ERK2 by cyclic strain, but a FAK mutant at the COOH terminal (Y925F) prevented the strain-induced activation of ERK2. Although the Y397F-Y576F Y577F FAK construct exhibited less basal FAK-Tyr 925 phosphorylation under static conditions, it nevertheless exhibited increased FAK-Tyr 925 phosphorylation in response to strain. These results suggest that repetitive deformation stimulates intestinal epithelial motility across fibronectin in a manner that requires both Src activation and a novel Src-independent FAK-Tyr 925-dependent pathway that activates ERK. This pathway may be an important target for interventions to promote mucosal healing in settings of intestinal ileus or fasting. PMID- 18400994 TI - Reciprocal changes in calcification of the gastrolith and cuticle during the molt cycle of the red claw crayfish Cherax quadricarinatus. AB - Mobilization of calcium during the molt cycle from the cuticle to transient calcium deposits is widely spread in crustaceans. The dynamics of calcium transport to transient calcium deposits called gastroliths and to the cuticle over the course of the molt cycle were studied in the crayfish Cherax quadricarinatus. In this species, calcium was deposited in the gastroliths during premolt and transported back to the cuticle during postmolt, shown by digital X ray radiograph analysis. The predominant mineral in the crayfish is amorphous calcium carbonate embedded in an organic matrix composed mainly of chitin. Scanning electron micrographs of the cuticle during premolt showed that the endocuticle and parts of the exocuticle were the source of most of the labile calcium, while the epicuticle did not undergo degradation and remained mineralized throughout the molt cycle. The gastroliths are made of concentric layers of amorphous calcium carbonate intercalated between chitinous lamella. Measurements of pH and calcium levels during gastrolith deposition showed that calcium concentrations in the gastroliths, stomach, and muscle were about the same (10 to 11 mmol l(-1)). On the other hand, pH varied greatly, from 8.7+/-0.15 in the gastrolith cavity through 7.6+/-0.2 in muscle to 6.9+/-0.5 in the stomach. PMID- 18400995 TI - Endosymbionts of Siboglinum fiordicum and the phylogeny of bacterial endosymbionts in Siboglinidae (Annelida). AB - Siboglinid worms are a group of gutless marine annelids that are nutritionally dependent upon endosymbiotic bacteria. Four major groups of siboglinids are known vestimentiferans, moniliferans, Osedax spp. and frenulates. Although endosymbionts of vestimentiferans and Osedax spp. have been previously characterized, little is currently known about endosymbiotic bacteria associated with frenulate and moniliferan siboglinids. This is particularly surprising given that frenulates are the most diverse and widely distributed group of siboglinids. Here, we molecularly characterize endosymbiotic bacteria associated with the frenulate siboglinid Siboglinum fiordicum by using 16S rDNA ribotyping in concert with laser-capture microdissection (LCM). Phylogenetic analysis indicates that at least three major clades of endosymbiotic gamma-proteobacteria associate with siboglinid annelids, with each clade corresponding to a major siboglinid group. S. fiordicum endosymbionts are a group of gamma-proteobacteria that are divergent from bacteria associated with vestimentiferan or Osedax hosts. Interestingly, symbionts of S. fiordicum, from Norway, are most closely related to symbionts of the frenulate Oligobrachia mashikoi from Japan, suggesting that symbionts of frenulates may share common evolutionary history or metabolic features. PMID- 18400996 TI - Tunic phagocytes are involved in allorejection reaction in the colonial tunicate Aplidium yamazii (Polyclinidae, Ascidiacea). AB - The colonial ascidian Aplidium yamazii exhibited an allorejection reaction when two allogeneic colonies were brought into contact at their growing edges or at artificial cut surfaces. This species has no vascular network in the tunic, unlike the botryllid ascidians, which have a vascular network throughout the colony's common tunic. In the allorejection reaction induced by contact at the growing edges, some small, hard-packed tunic masses were formed at the contact points. Histological and electron microscopic investigation of these tunic masses revealed that they contained aggregates of tunic cells, with tunic phagocytes being the major cell type present. Some of the tunic phagocytes in these tunic masses appeared to be disintegrating. When allogeneic colonies were placed in contact at their artificial cut surfaces, the colonies partially fused, then separated. In this allorejection reaction, some loosely packed tunic masses remained in the gap between the two withdrawn colonies. These results strongly suggest that the tunic phagocytes are likely to be the major effector cells in the allorejection reaction. We also propose that the tunic phagocytes are not only the effector cells in the allorejection reaction but also bear the sites of allorecognition. PMID- 18400997 TI - Origin and expansion of foliated microstructure in pteriomorph bivalves. AB - The ultrastructure of the calcitic prisms of the prismatic shell layers of pteriomorph bivalves was examined by scanning electronic microscopy and diffraction techniques. Results indicate that the internal structure of the prisms is noticeably different among taxa. In species belonging to the families Pinnidae, Pteriidae, and Isognomonidae (Pterioida), prisms are built up with nanometric calcite crystals. On the other hand, Pectinidae, Propeamussliidae, Anomiidae (order Pectinoida) and the Ostreidae (Ostreoida) have prisms constituted by calcitic laths with micrometric size. These laths are indistinguishable from those constituting the foliated microstructure. In almost all cases, there is mineral continuity from the prisms to the underlying foliated layer, as confirmed by X-ray texture analyses. These findings corroborate a previous assumption that the foliated microstructure derived from calcitic prisms, particularly from those with internal foliated structure. The appearance of the foliated microstructure facilitated drastic mineralogical and microstructural changes in pteriomorph shells-for example, the development of rigid shell margins and the production of largely calcitic shells. Such changes have, no doubt, contributed to the evolutionary success of the groups, which have shown a pronounced diversification over time. PMID- 18400998 TI - Laboratory observations on the feeding behavior and feeding rate of the nemertean Procephalothrix simulus. AB - The feeding behavior of the nemertean Procephalothrix simulus and the effects of extrinsic factors on the feeding rates of this nemertean were experimentally studied. Procephalothrix simulus showed a typical macrophagous feeding strategy and consumed various food items. When capturing freshwater Tubifex sp., the nemerteans successfully ingested prey in all attacks, and they did not evert the proboscis in 25% of capture events. When capturing marine Saccocirrus gabrillae, the nemerteans failed to consume prey in more than half of the attacks, and they always everted the proboscis. There was a positive relationship between nemertean body weight and the rate of successful attacks, and a negative relationship between nemertean body weight and the duration of feeding events. The feeding rate of P. simulus increased when the temperature was raised from 5 degrees C to 30 degrees C but was significantly inhibited at 32 degrees C. Food intake was significantly reduced in media diluted to a practical salinity of 20 and 10 and in medium with the salinity elevated to 45. Dark conditions induced higher food intake, but prey density had no significant effect on feeding rate. These results suggest that P. simulus is a predator successfully adapted to the variable environmental conditions of the intertidal habitat. PMID- 18400999 TI - Phylogenetic analysis of caprellid and corophioid amphipods (Crustacea) based on the 18S rRNA gene, with special emphasis on the phylogenetic position of Phtisicidae. AB - Members of the amphipod suborder Caprellidea exhibit degenerated abdomens and pereopods 3 and 4. Some genera of Podoceridae (Gammaridea, Corophioidea) such as Dulichia also show reduced abdomens and pereopods and thus are generally regarded as a sister group of the Caprellidea. In addition, one of the caprellid families, the Caprogammaridae, exhibits abdominal segments that are similar to those of the podocerids, as well as rudimentary pereopods 3 and 4, which are more consistent with those of other caprellids. Therefore, an evolutionary scheme has been suggested on the basis of the gradual degeneration of the pereopods and abdomen: [Dulichia, (caprogammarids, caprellids)]. However, the Phtisicidae (Caprellidea) contradict this hypothesis because they exhibit well-developed pereopods 3 and 4, along with degenerated abdomens. Therefore, previous studies have suggested that the Phtisicidae and other caprellids may be polyphyletic. We examined the phylogenetic position of the Phtisicidae and other caprellid amphipods, using 18S rRNA gene sequence data. The results strongly indicate that the Phtisicidae and other caprellid families form a monophyletic clade. However, a close phylogenetic relationship among Dulichia (Corophioidea) and taxa belonging to the Caprellidea was not definitively supported. This study is the first to use molecular data to investigate the phylogenetic relationships among the Caprellidea. PMID- 18401000 TI - On fertilization in Chaetopleura apiculata and selected Chitonida. AB - Early events of fertilization are described in Chaetopleura apiculata and other selected Chitonida. C. apiculata egg hulls are elaborated into multi-branched spines with interlocking polygonal bases. Around the perimeter of each base are a series of open pores, ranging in size from 0.1-0.5 microm, which permit sperm direct access to the vitelline layer. In Callochitonidae (Chitonida) even larger pores occur in egg jelly coats, but this is considered to be the plesiomorphic condition, found also in Lepidopleurida such as Deshayesiella curvata. Other Chitonina, such as Rhyssoplax tulipa and Acanthopleura granulata, have a continuous outer dense layer that lacks pores and must be digested by penetrating sperm. Fertilization in Chitonida is unique and involves injection of chromatin into the egg via a narrow tubular nuclear extension that appears to exclude other sperm organelles, including mitochondria, centrioles, and flagellum. New evidence from studies of fertilization in Mopalia muscosa (Chitonida: Acanthochitonina) supports this hypothesis. This type of fertilization implies maternal inheritance of both mitochondria and centrioles, which is highly unusual, because in most animals one sperm centriole assists movements of pronuclei and regulates organization of the mitotic spindle. This mechanism of fertilization is defined by a series of apomorphic characters that unify the order Chitonida. PMID- 18401001 TI - Patterns of male reproductive success in Crepidula fornicata provide new insight for sex allocation and optimal sex change. AB - The size-advantage model and sex-allocation theory are frequently invoked to explain the evolution and maintenance of sequential hermaphroditism in many taxa. A test of current theory requires quantitative estimates of reproductive success and knowledge of the relationship between reproduction and size for each gender. Reproductive success can be difficult to measure. In species where polyandry occurs, it can be quantified only by determining paternity of offspring. We employed microsatellite loci to establish paternity for 12 families of Crepidula fornicata, where a family is defined as a single female, her brood, and the males stacked on top of her. Genetic data were analyzed and paternity was assigned to a single potential father for more than 83% of the offspring tested. Estimates of reproductive success revealed that one male within the family fathered the majority of offspring and that he was usually the largest male and the one closest to the brooding female. The dominant male's success also tended to decrease as the number of mature males within the family increased. Our results suggest that sperm competition could be a driving force in determining male reproductive success and the timing of sex change in C. fornicata. PMID- 18401002 TI - Effect of chronic inhibition of converting enzyme on proximal tubule acidification. AB - The acute effect of angiotensin-converting enzyme inhibition (ACEi) on proximal convoluted tubule (PCT) function is well documented. However, the effect of chronic treatment is less known. The aim of this work was to evaluate the effect of chronic ACEi on PCT acidification (J(HCO(3)(-))). Rats received enalapril (10 mg.kg(-1).day(-1), added to the drinking water) during 3 mo. Micropuncture experiments were performed to measure the effect of chronic ACEi on J(HCO(3)(-)). Nitric oxide (NO.) synthesis in kidney cortex homogenates was assessed by quantifying the conversion of [(14)C]-L-arginine to [(14)C]-L-citrulline. Western blot analysis was performed to determine the abundances of V-H(+)ATPase and NHE3 isoform of the Na(+)/H(+) exchanger in proximal brush-border membrane vesicles (BBMV). Enalapril treatment induced an approximately 50% increase in J(HCO(3)( )). Luminal perfusion with ethyl-isopropyl amiloride (EIPA) 10(-4)M or bafilomycin 10(-6)M decreased J(HCO(3)(-)) by approximately 60% and approximately 30%, respectively, in both control and enalapril-treated rats. The effect of EIPA and bafilomycin on absolute J(HCO(3)(-)) was larger in enalapril-treated than in control rats. Acute inhibition of NO. synthesis with N(G)-nitro-L-arginine methyl ester abolished the enalapril-induced increase in J(HCO(3)(-)). Cortex homogenates from enalapril-treated rats displayed a 46% increase in nitric oxide synthase (NOS) activity compared with those from untreated animals. Enalapril treatment did not affect the abundances of NHE3 and V-H(+)ATPase in BBMV. Our results suggest that PCT acidification is increased during chronic ACEi probably due to an increase in NO. synthesis, which would stimulate Na(+)/H(+) exchange and electrogenic proton transport. PMID- 18401003 TI - ABCB- and ABCC-type transporters confer multixenobiotic resistance and form an environment-tissue barrier in bivalve gills. AB - Aquatic organisms and, in particular, filter feeders, such as mussels, are continuously exposed to toxicants dissolved in the water and, presumably, require adaptations to avoid the detrimental effects from such chemicals. Previous work indicates that activity of ATP-binding cassette (ABC) transporters protects mussels against toxicants, but the nature of these transporters and the structural basis of protection are not known. Here we meld studies on transporter function, gene expression, and localization of transporter protein in mussel gill tissue and show activity and expression of two xenobiotic transporter types in the gills, where they provide an effective structural barrier against chemicals. Activity of ABCB/MDR/P-glycoprotein and ABCC/MRP-type transporters was indicated by sensitivity of efflux of the test substrate calcein-AM to the ABCB inhibitor PSC-833 and the ABCC inhibitor MK-571. This activity profile is supported by our cloning of the complete sequence of two ABC transporter types from RNA in mussel tissue with a high degree of identity to transporters from the ABCB and ABCC subfamilies. Overall identity of the amino acid sequences with corresponding homologs from other organisms was 38-50% (ABCB) and 27-44% (ABCC). C219 antibody staining specific for ABCB revealed that this transporter was restricted to cells in the gill filaments with direct exposure to water flow. Taken together, our data demonstrate that ABC transporters form an active, physiological barrier at the tissue-environment interface in mussel gills, providing protection against environmental xenotoxicants. PMID- 18401006 TI - Adenylate cycalse toxin of Bordetella pertussis inhibits TLR-induced IRF-1 and IRF-8 activation and IL-12 production and enhances IL-10 through MAPK activation in dendritic cells. AB - Adenylate cyclase toxin (CyaA) of Bordetella pertussis binds to CD11b/CD18 on macrophages and dendritic cells (DC) and confers virulence to the bacteria by subverting innate immune responses of the host. We have previously demonstrated that CyaA promotes the induction of IL-10-secreting regulatory T cells in vivo by modulating DC activation. Here, we examine the mechanism of immune subversion, specifically, the modulation of TLR signaling pathways in DC. We found that CyaA synergized with LPS to induce IL-10 mRNA and protein expression in DC but significantly inhibited IL-12p70 production. CyaA enhanced LPS-induced phosphorylation of p38 MAPK and ERK in DC, and inhibitors of p38 MAPK, MEK, or NF kappaB suppressed IL-10 production in response to LPS and CyaA. However, inhibition of p38 MAPK, MEK, and NF-kappaB did not reverse the inhibitory effect of CyaA on TLR agonist-induced IL-12 production. Furthermore, CyaA suppression of IL-12 was independent of IL-10. In contrast, CyaA suppressed LPS- and IFN-gamma induced IFN-regulatory factor-1 (IRF-1) and IRF-8 expression in DC. The modulatory effects of CyaA were dependent on adenylate cyclase activity and induction of intracellular cAMP, as an enzyme-inactive mutant of CyaA failed to modulate TLR-induced signaling in DC, whereas the effects of the wild-type toxin were mimicked by stimulation of the DC with PGE2. Our findings demonstrate that CyaA modulates TLR agonist-induced IL-10 and IL-12p70 production in DC by, respectively, enhancing MAPK phosphorylation and inhibiting IRF-1 and IRF-8 expression and that this is mediated by elevation of intercellular cAMP concentrations. PMID- 18401004 TI - Connexin 40 and ATP-dependent intercellular calcium wave in renal glomerular endothelial cells. AB - Endothelial intracellular calcium ([Ca(2+)](i)) plays an important role in the function of the juxtaglomerular vasculature. The present studies aimed to identify the existence and molecular elements of an endothelial calcium wave in cultured glomerular endothelial cells (GENC). GENCs on glass coverslips were loaded with Fluo-4/Fura red, and ratiometric [Ca(2+)](i) imaging was performed using fluorescence confocal microscopy. Mechanical stimulation of a single GENC caused a nine-fold increase in [Ca(2+)](i), which propagated from cell to cell throughout the monolayer (7.9 +/- 0.3 microm/s) in a regenerative manner (without decrement of amplitude, kinetics, and speed) over distances >400 microm. Inhibition of voltage-dependent calcium channels with nifedipine had no effect on the above parameters, but the removal of extracellular calcium reduced Delta[Ca(2+)](i) by 50%. Importantly, the gap junction uncoupler alpha glycyrrhetinic acid or knockdown of connexin 40 (Cx40) by transfecting GENCs with Cx40 short interfering RNA (siRNA) almost completely eliminated Delta[Ca(2+)](i) and the calcium wave. Breakdown of extracellular ATP using a scavenger cocktail (apyrase and hexokinase) or nonselective inhibition of purinergic P2 receptors with suramin, had similar blocking effects. Scraping cells off along a line eliminated physical contact between cells but did not effect calcium wave propagation. Using an ATP biosensor technique, we detected a significant elevation in extracellular ATP (Delta = 76 +/- 2 microM) during calcium wave propagation, which was abolished by Cx40 siRNA treatment (Delta = 6 +/- 1 microM). These studies suggest that connexin 40 hemichannels and extracellular ATP are key molecular elements of the glomerular endothelial calcium wave, which may serve important juxtaglomerular functions. PMID- 18401008 TI - TSPY expression is variably altered in transgenic mice with testicular feminization. AB - TSPY (testis-specific protein, Y-encoded) genes are expressed in premeiotic germ cells and round spermatids. The topology and timing of TSPY expression, and also its homology to members of the TTSN-family, suggest that TSPY is a proliferation factor for germ cells. There is also evidence for a role of TSPY in the aetiology of testis cancer. TSPY is a candidate for GBY, the elusive gonadoblastoma locus on the human Y chromosome, which is thought to predispose dysgenetic gonads of 46, XY sex-reversed females to develop gonadoblastoma. We have previously generated a TSPY transgenic mouse line (Tg(TSPY)9Jshm) that carries approximately 50 copies of the human TSPY gene on the mouse Y chromosome. In order to elucidate TSPY expression under complete androgen insensitivity and to investigate a possible role of TSPY in gonadal tumorigenesis, we have now generated sex reversed TSPY transgenic Ar(Tfm) mice hemizygous for the X-linked testicular feminization mutation (Ar(Tfm)). We can show that the TSPY transcript is aberrantly spliced in the testes of TSPY-Ar(Tfm) mice, and that TSPY expression is upregulated by androgen insensitivity in some but not all animals. TSPY transgenic mice showed significantly increased testes weights. In one TSPY transgenic Ar(Tfm) animal, spermatogenesis proceeded beyond meiotic prophase. No tumors of germ cell origin were found in the testes of TSPY-Ar(Tfm) mice. Five out of 46 TSPY transgenic Ar(Tfm) mice, and 3 out of 31 age-related NMRI-Ar(Tfm) controls developed Leydig cell tumors, whereas none of the age-matched Ar(Tfm) mice (n=44) on a wild type background were affected by Leydig cell tumorigenesis. PMID- 18401005 TI - Acute alcohol intoxication increases atrogin-1 and MuRF1 mRNA without increasing proteolysis in skeletal muscle. AB - Acute alcohol intoxication decreases muscle protein synthesis, but there is a paucity of data on the ability of alcohol to regulate muscle protein degradation. Furthermore, various types of atrophic stimuli appear to regulate ubiquitin proteasome-dependent proteolysis by increasing the muscle-specific E3 ligases atrogin-1 and MuRF1 (i.e., "atrogenes"). Therefore, the present study was designed to test the hypothesis that acute alcohol intoxication increases atrogene expression leading to an elevated rate of muscle protein breakdown. In male rats, the intraperitoneal injection of alcohol dose- and time-dependently increased atrogin-1 and MuRF1 mRNA in gastrocnemius, the latter of which was most pronounced. A comparable change was absent in the soleus and heart. The ability of in vivo-administered ethanol to increase atrogene expression was independent of the route of alcohol administration (intraperitoneal vs. oral), as well as of nutritional status (fed vs. fasted) and gender (male vs. female). The increase in atrogin-1 and MuRF1 was independent of alcohol metabolism, and the overproduction of endogenous glucocorticoids and could not be prevented by maintaining the circulating concentration of insulin-like growth factor-I. Despite marked changes in atrogene expression, acute alcohol in vivo did not alter the release of either 3-methylhistidine (MH) or tyrosine from the isolated perfused hindlimb, suggesting that the rate of muscle proteolysis remains unchanged. Moreover, alcohol did not increase the directly determined rate of protein degradation in isolated epitrochlearis muscles or cultured myocytes. Finally, no increase in atrogene expression or 3-MH release was detected in muscle from rats fed an alcohol-containing diet. Our results indicate that although acute alcohol intoxication increases atrogin-1 and MuRF1 mRNA preferentially in fast-twitch skeletal muscle, this change was not associated with increased rates of muscle proteolysis. Therefore, the loss of muscle mass/protein in response to chronic alcohol abuse appears to result primarily from a decrement in muscle protein synthesis, not an increase in degradation. PMID- 18401007 TI - Mice deficient for a small cluster of Piwi-interacting RNAs implicate Piwi interacting RNAs in transposon control. AB - The mammalian testis expresses a class of small noncoding RNAs that interact with mammalian PIWI proteins. In mice, the PIWI-interacting RNAs (piRNAs) partner with mammalian PIWI proteins, PIWIL1 and PIWIL2, also known as MIWI and MILI, to maintain transposon silencing in the germline genome. Here, we demonstrate that inactivation of Nct1/2, two noncoding RNAs encoding piRNAs, leads to derepression of LINE-1 (L1) but does not affect mouse viability, spermatogenesis, testicular gene expression, or fertility. These findings indicate that piRNAs from a cluster on chromosome 2 are necessary to maintain transposon silencing. PMID- 18401009 TI - Estimating mitochondrial DNA content of chinook salmon spermatozoa using quantitative real-time polymerase chain reaction. AB - Animal mitochondrial DNA (mtDNA) is predominantly inherited maternally. Various mechanisms to avoid the transmission of paternal mtDNA to offspring have been proposed, including the dilution of paternal mtDNA by maternal mtDNA in the zygote. The effectiveness of dilution as a barrier will be determined by the number of mtDNA molecules contributed by each parental gamete, and is expected to be highly variable among different taxa due to interspecific differences in mating systems and gamete investment. Estimates of this ratio are currently limited to few mammalian species, and data from other taxa are therefore needed to better understand the mechanisms of mitochondrial inheritance. The present study estimates mtDNA content in salmon sperm, the first nonmammalian vertebrate to be examined. Although highly divergent, it appears that the mtDNA content may be conserved within vertebrate taxa, indicating that the reduction of mtDNA is a key factor of spermatogenesis to ensure mitochondrial functionality on the one hand, and to avoid paternal leakage at a significant or detectable level on the other hand. We employ quantitative real-time PCR (Q-PCR) and demonstrate the accuracy and high reproducibility of our experiments. Furthermore, we compare and evaluate two standard approaches used for the quantification of genes, Q-PCR and blotting methods, in regard to their utility in the accurate quantification of mitochondrial genes. PMID- 18401010 TI - Hamster sperm capacitation: role of pyruvate dehydrogenase A and dihydrolipoamide dehydrogenase. AB - Recently, we demonstrated that pyruvate dehydrogenase A2 (PDHA2) is tyrosine phosphorylated in capacitated hamster spermatozoa. In this report, using bromopyruvate (BP), an inhibitor of PDHA, we demonstrated that hamster sperm hyperactivation was blocked regardless of whether PDHA was inhibited prior to or after the onset of hyperactivation, but the acrosome reaction was blocked only if PDHA was inhibited prior to the onset of the acrosome reaction. Further, inhibition of PDHA activity did not inhibit capacitation-associated protein tyrosine phosphorylation observed in hamster spermatozoa. It is demonstrated that the essentiality of PDHA for sperm capacitation is probably dependent on its ability to generate effectors of capacitation such as reactive oxygen species (ROS) and cAMP, which are significantly decreased in the presence of BP. MICA (5 methoxyindole-2-carboxylic acid, a specific inhibitor of dihydrolipoamide dehydrogenase [DLD]), another component of the pyruvate dehydrogenase complex (PDHc), also significantly inhibited ROS generation and cAMP levels thus implying that these enzymes of the PDHc are required for ROS and cAMP generation. Furthermore, dibutryl cyclic adenosine monophosphate could significantly reverse the inhibition of hyperactivation observed in the presence of BP and inhibition of acrosome reaction observed in the presence of BP or MICA. The calcium ionophore, A23187, could also significantly reverse the inhibitory effect of BP and MICA on sperm acrosome reaction. These results establish that PDHA is required for hamster sperm hyperactivation and acrosome reaction, and DLD is required for hamster acrosome reaction. This study also provides evidence that ROS, cAMP, and calcium are involved downstream to PDHA. PMID- 18401011 TI - Heredity--venturing beyond genetics. AB - Our knowledge of heredity has recently undergone major upheaval. Heredity transmits considerably more than just genetic elements. First, the oocyte is full of maternal cytoplasmic components that subsequently are present in each new cell. Second, maternal cells can pass to the progeny, where they remain active into adult life (microchimerism). Here, we examine the notion that the transmission of characters involves at least two processes in addition to that of mendelian heredity, long considered to be the only hereditary mechanism. These processes all involve epigenetic processes, including the transmission of macromolecules, subcellular organelles, and living cells solely from the mother to her offspring, whether female or male, during pregnancy and lactation. We postulate that cytoplasmic heredity and maternal transmission of cells leading to a long-term state of microchimerism in progeny are two good examples of matrilineal, nonmendelian heredity. A mother's important contribution to the development and health of her progeny seems to possess many uncharted depths. PMID- 18401012 TI - Extracellular adenosine 5'-triphosphate alters motility and improves the fertilizing capability of mouse sperm. AB - Extracellular adenosine 5'-triphosphate (ATPe) treatment of human sperm has been implicated in improving in vitro fertilization (IVF) results. We used the mouse model to investigate mechanisms of action of ATPe on sperm. ATPe treatment significantly enhanced IVF success as indicated by both rate of pronuclear formation and percentage cleavage to the 2-cell stage. However, ATPe did not increase the percentage of sperm undergoing spontaneous acrosomal exocytosis nor change the pattern of protein tyrosine phosphorylation normally observed in capacitated sperm. ATPe altered sperm motility parameters; in particular, both noncapacitated and capacitated sperm swam faster and straighter. The percentage of hyperactivated sperm did not increase in capacitated ATPe-treated sperm compared to control sperm. ATPe induced a rapid increase in the level of intracellular calcium that was inhibited by two distinct P2 purinergic receptor inhibitors, confirming that these receptors have an ionotropic role in sperm function. The observed motility changes likely explain, in part, the improved fertilizing capability when ATPe-treated sperm were used in IVF procedures and suggest a mechanism by which ATPe treatment may be beneficial for artificial reproductive techniques. PMID- 18401013 TI - RAB2A: a major subacrosomal protein of bovine spermatozoa implicated in acrosomal biogenesis. AB - The perinuclear theca (PT) of mammalian sperm is a unique subcellular structure encapsulating the nucleus. Compositionally, the PT is made up of at least six prominent polypeptides (60, 36, 31, 28, 24, and 15 kDa), of which only two have been sequence identified, as well as many less prominent ones. As an ongoing process in unveiling the protein composition of the PT, we have uncovered the sequence identity of the prominent 24-kDa polypeptide (PT24). Initial N-terminal sequence analysis obtained by Edman degradation suggested that PT24 is a RAB2 protein. This was corroborated by mass spectrometric analyses of trypsin-digested fragments of PT24, identifying RAB2A of the RAB2 subfamily as the best sequence match. Quadrapole/time-of-flight analysis identified 72%% sequence coverage between PT24 and bull, human, mouse, or rabbit RAB2A. Since a genome search only identified two RAB2 subfamily members, RAB2A and RAB2B, the 72%% sequence coverage of PT24 provides assurance that this protein is RAB2A and not a new RAB2 subfamily member. Furthermore, commercial RAB2A antibodies, raised against oligopeptide fragments in the unique C-terminal region of RAB2A, specifically labeled PT24 on Western blot analysis of PT extracts. These anti-RAB2A antibodies, along with immune serum that we raised and affinity purified against isolated PT24, demonstrated at both light and electron microscope levels that RAB2 is associated with the periphery of the growing proacrosomic and acrosomic vesicles in the Golgi and cap phases of spermiogenesis and consequently assembled as part of the PT. This pattern of subacrosomal assembly is reminiscent of the pathway used by SubH2Bv (PT15), another prominent and exclusive subacrosomal protein, indicating a common route for subacrosomal-PT assembly. Traditionally somatic RAB2 proteins are involved in vesicular transport between the endoplasmic reticulum and the cis-side of the Golgi apparatus. Our study suggests an unprecedented direction of RAB2A-mediated vesicular transport in spermatids during acrosomal biogenesis, from the trans-side of the Golgi apparatus to the nuclear envelope. PMID- 18401014 TI - Coexpression of adrenomedullin and its receptors in the reproductive system of the rat: effects on steroid secretion in rat ovary. AB - The present study demonstrates the expression of adrenomedullin (ADM) in the reproductive system of the female rat and its effect on the secretion of estradiol and progesterone. Ovarian ADM and Adm mRNA levels were decreased at estrus, whereas oviductal Adm mRNA levels were low at proestrus. Both tissues were shown to coexpress mRNAs encoding the calcitonin receptor-like receptor and receptor activity-modifying protein 1 (Ramp1), Ramp2, and Ramp3. Gel filtration chromatography of ovarian extracts showed two peaks, with the predominant one eluting at the position of authentic rat ADM (1-50) at estrus and at the position of ADM precursor at diestrus. Positive ADM immunostaining was localized in the granulosa and theca cells of the follicle and corpora lutea of the ovary. Adrenomedullin inhibited FSH-induced estradiol secretion in 2-day-old follicles and also suppressed eCG-stimulated progesterone release in corpora lutea. The inhibitory effect of ADM on the follicles and the corpora lutea was abolished by calcitonin gene-related peptide (8-37) and ADM (22-52), respectively. The presence of ADM and the gene expression of ADM and its receptor components in the female reproductive system suggest a paracrine effect of ADM on ovarian steroidogenesis. PMID- 18401016 TI - Cutaneous pharmacodynamics of a toll-like receptor 7 agonist, 852A, in humans. AB - 852A is a specific toll-like receptor 7 (TLR7) agonist. Thirty-two healthy adults (8 subjects per group) received two 1-g topical applications over 400 cm(2), separated by >or= 5 days, of 852A 0.01% followed by vehicle, vehicle followed by 852A 0.1%, 852A 0.3% followed by vehicle, or vehicle followed by 852A 1.0%. Systemic absorption was minimal as 852A was not quantifiable in any serum sample up to 24 hours postadministration and was only quantifiable at 24 hours in the urine of 4 of 8 subjects after application of 852A 1.0%. No systemic adverse events were associated with drug treatment. Gene expression analysis from application site biopsies showed a >or=2-fold increase in expression for 40 genes in at least 2 subjects. CXCL9/MIG (8/32 subjects), CCL2/MCP1 (7/32), and OAS3 (5/32) were most frequently increased, followed by other type I interferon inducible genes. Cluster analysis of the genes with a >or=2-fold increase did not reveal a definitive pattern with respect to 852A concentration or time of biopsy. Overall, single topical application of 852A up to 1.0% was well tolerated. Data gathered from these subjects are suggestive that 852A can produce increases in local gene expression consistent with TLR7 stimulation. PMID- 18401015 TI - Disease progression or pseudoprogression after concomitant radiochemotherapy treatment: pitfalls in neurooncology. AB - Although radionecrosis has been exhaustively described in depth in the neurooncological literature, its diagnosis is still a challenging issue because its radiological pattern is frequently indistinguishable from that of tumor recurrence. This review discusses the causes of radionecrosis and the potential effect of adjuvant chemotherapy concomitant with radiotherapy on its rate and onset. The potential pitfalls in clinical studies attempting to make a differential diagnosis between radionecrosis and disease progression are also discussed. PMID- 18401017 TI - Population pharmacokinetics of infliximab in patients with ankylosing spondylitis. AB - The population pharmacokinetics of infliximab were characterized in patients with active ankylosing spondylitis (n = 274). Serum infliximab concentration data, from a 2-year period, were analyzed using NONMEM. A 2-compartment linear pharmacokinetic model was chosen to describe the pharmacokinetic characteristics of infliximab in serum. Population estimates (typical value +/- standard error) were obtained from the final covariate model: clearance (CL: 0.273 +/- 0.007 L/day), volume of distribution in the central compartment (V(1): 3.06 +/- 0.057 L), intercompartment clearance (Q: 1.72 +/- 0.48 L/day), and volume of distribution in the peripheral compartment (V(2): 2.94 +/- 0.17 L). Interindividual variability for CL and V(1) was 34.1% and 17.5%, respectively. White blood cell count at baseline and the antibody-to-infliximab status were significant covariates to CL; body surface area and sex were significant covariates to V(1). The CL for patients with a positive antibody-to-infliximab status was estimated to be 41.9% to 76.7% higher than for the remaining patients. Other covariates (baseline disease activity and the concomitant medication use of prednisolone, omeprazole, nonsteroidal anti-inflammatory drugs, or analgesics) did not affect infliximab pharmacokinetics. The development of antibodies to infliximab was associated with accelerated infliximab clearance and may represent a potential underlying mechanism for an inadequate response, or loss of response, to infliximab treatment. PMID- 18401018 TI - Total cholesterol and the risk of Parkinson disease. AB - OBJECTIVE: To examine the association between serum total cholesterol at baseline and the risk of Parkinson disease (PD). METHODS: Study cohorts included 24,773 Finnish men and 26,153 women aged 25 to 74 years without a history of PD and stroke at baseline. Hazard ratios (HRs) of incident PD were estimated for different levels of total cholesterol. RESULTS: During a mean follow-up period of 18.1 years, 321 men and 304 women developed incident PD. After adjustment for confounding factors (age, study years, body mass index, systolic blood pressure, education, leisure-time physical activity, smoking, alcohol consumption, coffee and tea consumption, and history of diabetes), the HRs of PD at different levels of total cholesterol (<5, 5-5.9, 6-6.9, and >or=7 mmol/L) were 1.00, 1.33, 1.53, and 1.84 (p for trend = 0.035) in men; 1.00, 1.55, 1.57, and 1.86 (p for trend = 0.113) in women; and 1.00, 1.42, 1.56, and 1.86 (p for trend = 0.002) in men and women combined (adjusted also for sex). In both sexes combined, the increased risk of PD associated with increasing levels of serum total cholesterol was present both in subjects aged 25-44 years and in subjects aged 45-54 years at baseline, and in never smokers and smokers; however, no association was found among subjects aged 55 years or older at baseline. CONCLUSION: This large prospective study suggests that high total cholesterol at baseline is associated with an increased risk of Parkinson disease. PMID- 18401019 TI - Results from a phase I safety trial of hAADC gene therapy for Parkinson disease. AB - BACKGROUND: In a primate model of Parkinson disease (PD), intrastriatal infusion of an adeno-associated viral (AAV) vector containing the human aromatic l-amino acid decarboxylase (hAADC) gene results in robust gene expression. After gene transfer, low doses of systemically administered l-dopa are converted to dopamine in the transduced striatal neurons, resulting in behavioral improvement without the side effects typically associated with higher doses of l-dopa. These studies led to the initiation of a phase I safety trial. Here we report the findings for the first cohort of five patients. METHODS: Patients with moderate to advanced PD received bilateral infusion of a low dose of the AAV-hAADC vector into the putamen. PET scans using the AADC tracer, 6-[18F]fluoro-l-m-tyrosine (FMT), were performed at baseline and at 1 and 6 months after infusion as an in vivo measure of gene expression. RESULTS: PET results showed an average 30% increase in FMT uptake (K(i)(c)) in the putamen after gene transfer. Preliminary analysis of clinical data indicates a modest improvement, but absence of a control and the nonblinded analyses make interpretation difficult. CONCLUSIONS: Thus far, this gene therapy approach has been well tolerated and shows PET evidence of sustained gene expression. These initial findings demonstrate the safety of the therapy; higher doses of adeno-associated viral vector containing the human aromatic l amino acid decarboxylase gene in the next cohort of patients may further increase dopamine production in the putamen and provide more profound clinical benefit. PMID- 18401020 TI - Impaired insulin secretion increases the risk of Alzheimer disease. AB - OBJECTIVE: Subjects with diabetes are reported to have an increased risk of dementia and cognitive impairment. However, the underlying causes remain unknown. We investigated the longitudinal associations between midlife insulin secretion, glucose metabolism, and the subsequent development of Alzheimer disease (AD) and dementia. METHODS: The population-based Uppsala Longitudinal Study of Adult Men started 1970 when the 2,322 participants were 50 years old. Investigation at baseline included determinations of acute insulin response and glucose tolerance using the IV glucose tolerance test and Homeostasis Model Assessment insulin resistance index. During a median follow up of 32 years, 102 participants were diagnosed with AD, 57 with vascular dementia, and 394 with any dementia or cognitive impairment. Associations were analyzed using Cox proportional hazard models. RESULTS: A low insulin response at baseline was associated with a higher cumulative risk of AD (hazard ratio for 1 SD decrease, 1.31; 95% CI, 1.10-1.56) also after adjustment for age, systolic blood pressure, body mass index, serum cholesterol, smoking, education level, and insulin resistance. This association was stronger in subjects without the APOE epsilon4 allele. Impaired glucose tolerance increased the risk of vascular dementia (hazard ratio for 1 SD decrease, 1.45; 95% CI, 1.05-2.00) but not AD. Impaired insulin secretion, glucose intolerance, and estimates of insulin resistance were all associated with higher risk of any dementia and cognitive impairment. CONCLUSIONS: In this longitudinal study, impaired acute insulin response at midlife was associated with an increased risk of Alzheimer disease (AD) up to 35 years later suggesting a causal link between insulin metabolism and the pathogenesis of AD. PMID- 18401021 TI - Plasma amyloid levels and the risk of AD in normal subjects in the Cardiovascular Health Study. AB - OBJECTIVES: To examine the association between incident Alzheimer disease (AD), and plasma A beta 1-40 and A beta 1-42 levels in normal and mild cognitive impairment (MCI) subjects in a subgroup of participants of the Cardiovascular Health Study Cognition Study. METHODS: We determined the plasma A beta 1-40 and A beta 1-42 levels of 274 nondemented subjects (232 normals and 42 with MCI) in 1998-1999 and repeated the measurements in 2002-2003. The mean age of the subjects at baseline was 79.3 +/- 3.6 years. We examined the association between A beta levels and incident AD over the ensuing 4.5 years, controlling for age, cystatin C level (marker of glomerular function), apolipoprotein E-4 allele, Modified-Mini-Mental State Examination scores, and MRI-identified infarcts. RESULTS: In an unadjusted prospective model in normal subjects, both A beta 1-40 and A beta 1-42 levels in 1998-1999 were associated with incident AD (n = 55) in 2002-2003 (longitudinal analysis). In the fully adjusted multivariate model, neither A beta 1-42 nor A beta 1-40 nor their ratio was associated with incident AD. However, adjustment had a very small effect on point estimates for A beta 1 42, from an odds ratio (OR) of 1.61 (p = 0.007) in the unadjusted model to an OR of 1.46 (p = 0.08) in the fully adjusted model. In 2002-2003 (cross-sectional analysis), only the unadjusted models showed that both peptides were associated with AD. CONCLUSIONS: Plasma A beta levels are affected by age and by systemic and CNS vascular risk factors. After controlling for these conditions, A beta-40 and A beta 1-42 are weak predictors of conversion to Alzheimer disease (AD) in normal subjects and are only weakly associated with AD in cross-sectional analysis. Consequently, plasma levels of A beta do not seem to be useful biomarkers for AD. PMID- 18401022 TI - Abnormal TDP-43 immunoreactivity in AD modifies clinicopathologic and radiologic phenotype. AB - BACKGROUND: TAR DNA-binding protein 43 (TDP-43) is one of the major disease proteins in frontotemporal lobar degeneration with ubiquitin immunoreactivity. Approximately one-fourth of subjects with pathologically confirmed Alzheimer disease (AD) have abnormal TDP-43 (abTDP-43) immunoreactivity. The aim of this study was to determine whether subjects with pathologically confirmed AD and abTDP-43 immunoreactivity have distinct clinical, neuropsychological, imaging, or pathologic features compared with subjects with AD without abTDP-43 immunoreactivity. METHODS: Eighty-four subjects were identified who had a pathologic diagnosis of AD, neuropsychometric testing, and volumetric MRI. Immunohistochemistry for TDP-43 was performed on sections of hippocampus and medial temporal lobe, and positive cases were classified into one of three types. Neuropsychometric data were collated and compared in subjects with and without abTDP-43 immunoreactivity. Voxel-based morphometry was used to assess patterns of gray matter atrophy in subjects with and without abTDP-43 immunoreactivity compared with age- and sex-matched controls. RESULTS: Twenty-nine (34%) of the 84 AD subjects had abTDP-43 immunoreactivity. Those with abTDP-43 immunoreactivity were older at onset and death and performed worse on the Clinical Dementia Rating scale, Mini-Mental State Examination, and Boston Naming Test than subjects without abTDP-43 immunoreactivity. Subjects with and without abTDP-43 immunoreactivity had medial temporal and temporoparietal gray matter loss compared with controls; however, those with abTDP-43 immunoreactivity showed greater hippocampal atrophy. Multivariate logistic regression adjusting for age at death demonstrated that hippocampal sclerosis was the only pathologic predictor of abTDP-43 immunoreactivity. CONCLUSIONS: The presence of abnormal TDP 43 immunoreactivity is associated with a modified Alzheimer disease clinicopathologic and radiologic phenotype. PMID- 18401024 TI - Rufinamide for generalized seizures associated with Lennox-Gastaut syndrome. AB - BACKGROUND: Lennox-Gastaut syndrome is a catastrophic pediatric epilepsy syndrome characterized by multiple types of treatment-resistant seizures and high rates of seizure-related injury. Current available treatments are inadequate, leaving patients with few treatment options and opportunities. METHODS: We conducted a double-blind, randomized, placebo-controlled trial of the antiepileptic drug rufinamide in patients with Lennox-Gastaut syndrome. Eligible patients between 4 and 30 years of age had multiple types of seizures (including tonic-atonic and atypical absence seizures) with a minimum of 90 seizures in the month before baseline and a recent history of a slow spike-and-wave pattern on EEG. RESULTS: After a 28-day baseline period, 139 eligible patients were randomized; 138 patients received either rufinamide (n = 74) or placebo (n = 64) in addition to their other antiepileptic drugs. The median percentage reduction in total seizure frequency was greater in the rufinamide therapy group than in the placebo group (32.7% vs 11.7%, p = 0.0015). There was a difference (p < 0.0001) in tonic-atonic ("drop attack") seizure frequency with rufinamide (42.5% median percentage reduction) vs placebo (1.4% increase). The rufinamide group had a greater improvement in seizure severity (p = 0.0041) and a higher 50% responder rate compared with placebo for total seizures (p = 0.0045) and tonic-atonic seizures (p = 0.002). The common adverse events (reported by >or=10% of patients receiving rufinamide) were somnolence (24.3% with rufinamide vs 12.5% with placebo) and vomiting (21.6% vs 6.3%). CONCLUSIONS: Rufinamide was an effective and well tolerated treatment for seizures associated with Lennox-Gastaut syndrome. PMID- 18401023 TI - APOE epsilon 4 allele predicts faster cognitive decline in mild Alzheimer disease. AB - OBJECTIVE: To determine whether APOE epsilon 4 predicts rate of cognitive change in incident and prevalent Alzheimer disease (AD). METHODS: Individuals were recruited from two longitudinal cohort studies-the Washington Heights and Inwood Columbia Aging Project (WHICAP; population-based) and the Predictors Study (clinic-based)--and were followed for an average of 4 years. Three samples of participants diagnosed with AD, with diverse demographic characteristics and baseline cognitive functioning, were studied: 1) 199 (48%) of the incident WHICAP cases; 2) 215 (54%) of the prevalent WHICAP cases; and 3) 156 (71%) of the individuals diagnosed with AD in the Predictors Study. Generalized estimating equations were used to test whether rate of cognitive change, measured using a composite cognitive score in WHICAP and the Mini-Mental State Examination in Predictors, varied as a function of epsilon 4 status in each sample. RESULTS: The presence of at least one epsilon 4 allele was associated with faster cognitive decline in the incident population-based AD group (p = 0.01). Parallel results were produced for the two prevalent dementia samples only when adjusting for disease severity or excluding the most impaired participants from the analyses. CONCLUSION: APOE epsilon 4 may influence rate of cognitive decline most significantly in the earliest stages of Alzheimer disease. PMID- 18401025 TI - Silver syndrome variant of hereditary spastic paraplegia: A locus to 4p and allelism with SPG4. AB - OBJECTIVE: To perform a clinical and genetic study of two large Italian families (RM-36 and RM-51) showing the cardinal clinical features of Silver syndrome (SS), a rare dominantly inherited form of hereditary spastic paraplegia (HSP) complicated by amyotrophy of the small hand muscles. METHODS: Clinical assessment including neurophysiologic, neuropsychological, and neuroimaging evaluations. Genetic studies included linkage and sequence analyses. RESULTS: Using a genome wide survey in the RM-36 family, a novel locus (SPG38) has been identified and mapped within the 13.1-cM region on chromosome 4p16-p15 between markers D4S432 and D4S1599. The RM-51 family was linked to the SPG4 locus at 2p21-p24 and sequence analysis of SPG4 showed a novel frameshift mutation p.Asp321GlyfsX6. Clinical examination of the affected members carrying the mutation showed high frequency of additional clinical features including decreased vibration sense, pes cavus, temporal lobe epilepsy, and cognitive impairment. CONCLUSIONS: This study demonstrates evidence of a novel locus SPG38 for Silver syndrome (SS) and suggests that genetic defects in SPG4 might lead to broad clinical features overlapped with those of SS. PMID- 18401026 TI - Long-term efficacy of gamma knife radiosurgery in mesial temporal lobe epilepsy. AB - BACKGROUND: Gamma knife (GK) radiosurgery has been proposed as an alternative to classic microsurgery in mesial temporal lobe epilepsy (MTLE). Short-term follow up studies have reported encouraging results, but long-term efficacy is not known. OBJECTIVE: To report the efficacy and tolerance of GK radiosurgery in MTLE after a follow-up > 5 years. METHODS: Patients with a follow-up > 5 years presenting with MTLE and treated with a marginal dose of 24 Gy were included in the study. RESULTS: Fifteen patients were included. Eight were treated on the left side, and 7 were treated on the right. The mean follow-up was 8 years (range 6-10 years). At the last follow-up, 9 of 16 patients (60%) were considered seizure free (Engel Class I) (4/16 in Class IA, 5/16 in Class IB). Seizure cessation occurred with a mean delay of 12 months (+/- 3) after GK radiosurgery, often preceded by a period of increasing aura or seizure occurrence (6/15 patients). The mean delay of appearance of the first neuroradiologic changes was 12 months (+/- 4). Nine patients (60%) experienced mild headache and were placed on corticosteroid treatment for a short period. All patients who were initially seizure free experienced a relapse of isolated aura (10/15, 66%) or complex partial seizures (10/15, 66%) during antiepileptic drug tapering. Restoration of treatment resulted in good control of seizures. CONCLUSION: Gamma knife radiosurgery is an effective and safe treatment for mesial temporal lobe epilepsy. Results are maintained over time with no additional side effects. Long term results compare well with those of conventional surgery. PMID- 18401027 TI - Is muscle glycogenolysis impaired in X-linked phosphorylase b kinase deficiency? AB - OBJECTIVE: It is unclear to what extent muscle phosphorylase b kinase (PHK) deficiency is associated with exercise-related symptoms and impaired muscle metabolism, because 1) only four patients have been characterized at the molecular level, 2) reported symptoms have been nonspecific, and 3) lactate responses to ischemic handgrip exercise have been normal. METHODS: We studied a 50-year-old man with X-linked PHK deficiency using ischemic forearm and cycle ergometry exercise tests to define the derangement of muscle metabolism. We compared our findings with those in patients with McArdle disease and in healthy subjects. RESULTS: Sequencing of PHKA1 showed a novel pathogenic mutation (c.831G>A) in exon 7. There was a normal increase of plasma lactate during forearm ischemic exercise, but lactate did not change during dynamic, submaximal exercise in contrast to the fourfold increase in healthy subjects. Constant workload elicited a second wind in all patients with McArdle disease, but not in the patient with PHK deficiency. IV glucose administration appeared to improve exercise tolerance in the patient with PHK deficiency, but not to the same extent as in the patients with McArdle disease. Lipolysis was higher in the patient with PHK deficiency than in controls. CONCLUSION: These findings demonstrate that X linked PHK deficiency causes a mild metabolic myopathy with blunted muscle glycogen breakdown and impaired lactate production during dynamic exercise, which impairs oxidative capacity only marginally. The different response of lactate to submaximal and maximal exercise is likely related to differential activation mechanisms for myophosphorylase. PMID- 18401028 TI - The revised World Health Organization diagnostic criteria for polycythemia vera, essential thrombocytosis, and primary myelofibrosis: an alternative proposal. PMID- 18401029 TI - Knockout punches with a fistful of zinc fingers. PMID- 18401030 TI - Differences in spontaneous mutation frequencies as a function of environmental stress in soil fungi at "Evolution Canyon," Israel. AB - When various wild strains of Penicillium lanosum and Aspergillus niger were placed in the same mild laboratory environment, their frequencies of new spontaneous mutations were clearly related to whether they had been isolated from a region of high or low microclimatic stress. In the mild environment, the total frequencies of conidial color and morphological mutations in P. lanosum, summed over all relevant loci, ranged from 0.29% to 2.4% for six strains from the north facing, less stressful "European" slope (ES/NFS) of "Evolution Canyon" I, compared with 6.5-11.6% for five strains from the south-facing "African" slope (AS/SFS), which is a much more stressful environment, being harsher, drier, more fluctuating in temperature, and receiving up to eight times more UV radiation than the opposite slope. The corresponding figures for A. niger were 0.42-1.50% for three strains from the ES/NFS and 2.3-4.9% for six strains from the AS/SFS. The more mutagenic environment of the AS/SFS than of the ES/NFS means that, in Evolution Canyon, the mutation frequency differences between the very stressful environment and the less stressful environment are probably even larger than the 4- and 6-fold differences found here in a mild laboratory environment. The evidence from these two filamentous fungi, which have no sexual cycle, is that there are inherited differences in spontaneous mutation rates according to the levels of stress in the environment, and this feature may well be adaptive. Evolution Canyon I is at Nahal Oren, Mount Carmel, Israel. PMID- 18401031 TI - Worshiping false idols: the impact factor dilemma. PMID- 18401032 TI - Benign rolandic epilepsy -- perhaps not so benign: use of magnetic source imaging as a predictor of outcome. AB - The purpose of this study was to evaluate children with benign rolandic epilepsy, a childhood epilepsy characterized by centrotemporal/rolandic spike-wave discharges with infrequent partial seizures that may secondarily generalize. Recently, some investigators have questioned whether benign rolandic epilepsy is indeed "benign" or whether long-term cognitive outcome may be adversely affected. We initiated an ongoing study to identify children with benign rolandic epilepsy. The children were evaluated in the Texas Comprehensive Epilepsy Program using outpatient or continuous video-electroencephalographic monitoring, brain magnetic resonance imaging, magnetoencephalography, and neuropsychological testing. Neuropsychological testing revealed fine motor dysfunction, visuomotor integration deficits, dyscalculia, and/or expressive language deficits in all of the 9 patients evaluated, reaffirming that benign rolandic epilepsy is not necessarily a benign disorder. Our study shows a high concordance of motor and cognitive deficits in benign rolandic epilepsy, as others have previously suggested. Furthermore, magnetic source imaging shows a higher resolution of dipole localization compared with conventional electroencephalography, which may ultimately improve prediction of deficits. This reaffirms that magnetoencephalography is a valuable diagnostic tool in the evaluation of children with benign rolandic epilepsy. PMID- 18401033 TI - Handwriting performance in children with attention deficit hyperactivity disorder (ADHD). AB - Attention deficit hyperactivity disorder (ADHD) is the most common neurobehavioral condition of childhood. Consequences are multifaceted and include activity limitations in daily-living skills, academic challenges, diminished socialization skills, and motor difficulties. Poor handwriting performance is an example of an affected life skill that has been anecdotally observed by educators and clinicians for this population and can negatively impact academic performance and self-esteem. To guide health and educational service delivery needs, the authors reviewed the evidence in the literature on handwriting difficulties in children with ADHD. Existing evidence would suggest that children with ADHD have impaired handwriting performance, characterized by illegible written material and/or inappropriate speed of execution compared to children without ADHD. Studies with larger sample sizes using standardized measures of handwriting performance are needed to evaluate the prevalence of the problem and to better understand the nature of handwriting difficulties and their impact in this population. PMID- 18401034 TI - Psychosis with frontal lobe epilepsy responds to carbamazepine. AB - Frontal lobe epilepsy is an unusual form of complex partial seizures associated with behavioral changes including fear, agitation, kicking, and screaming. The seizures are often missed or mistaken for psychiatric problems or sleep disorders. We report 2 unusual patients presenting with psychosis who were found to have frequent frontal lobe seizures. Treatment of the seizures with carbamazepine resulted in complete resolution of their psychiatric symptoms. The relationship between frontal lobe seizures and psychosis is explored. PMID- 18401035 TI - Behavioral disorders in children with idiopathic intracranial hypertension. AB - The purpose of this research was to evaluate the association between idiopathic intracranial hypertension and behavior, attention, and learning abilities in children. Parents of school-age children with idiopathic intracranial hypertension were asked to fill out a questionnaire and to rank the child's behavioral patterns before and after the diagnosis and treatment of the disease. The questionnaire was based on Conners' test. Ten children were included in the study. Mean age at diagnosis was 11.5 years. Mean follow-up time was 25 months. Six patients (60%) met the definition of attention- and concentration-deficit disorders before diagnosis of idiopathic intracranial hypertension; 1 patient was treated with methylphenidate (Ritalin) before referral to eye examination. After the diagnosis was made and treatment was established, 5 patients (83%) reported an improvement in their attention and behavior. Of these 6 patients, 2 (33%) reported marked improvement. We conclude that attention- and concentration deficit disorder might be an early sign for pediatric idiopathic intracranial hypertension. Diagnosis and treatment of idiopathic intracranial hypertension in these children may improve the child's behavior, attention, and achievements in school, without the need to resort to other modes of therapy. PMID- 18401036 TI - Pediatric tanycytic ependymoma of the cauda equina: case report and review of the literature. AB - Tanycytic tumors of the cauda equina region are being increasingly reported. Such tumors can occur among pediatric patients and adults. An unusual case of tanycytic ependymoma located at the region of the cauda equina is reported in a girl. The clinical presentation, radiological features, intraoperative findings, and histopathologic features are described. The prognostic significance of differentiating such a neoplasm from other intramedullary tumors is noted. A heightened awareness of this pathologic entity among neurosurgeons and pathologists will help in diagnosing this neoplasm and in enabling better patient management. PMID- 18401037 TI - Multiple intracranial tuberculomas associated with partial status epilepticus and refractory infantile spasms. AB - We report the case of a 10 month-old girl presenting with multiple intracranial tuberculomas associated with partial status epilepticus and cerebrovascular accident in the left sylvian territory. She later developed paradoxical enlargement of the tuberculomas during antituberculous treatment and severe neurological sequelae with refractory infantile spasms. The development of infantile spasms in this context is particularly remarkable, and this case is illustrative of the complex interrelationship between intracranial lesions, partial status epilepticus, and infantile spasms. It also highlights the difficult diagnosis of cerebral tuberculomas in infants and further supports the need for continued vigilance in industrialized countries. PMID- 18401039 TI - Re: Blood levels of mercury are related to diagnosis of autism: a reanalysis of an important data set. PMID- 18401040 TI - Misphasing of ion motion in quadratic potential induced by space-periodic disturbance. AB - An ideally quadratic potential used in different types of ion mass analyzer such as Paul trap, Kingdon trap and quadratic field reflectron may be space periodically disturbed due to inaccuracy of fabrication and design features. If ion motion in such devices is computer-simulated, disturbances of potential may be caused by the peculiarity of the computation method. The problem investigated in this work is the effect that weak space-periodic disturbance of a quadratic potential takes on the ion motion in such a potential. The effect of the disturbance we considered is the misphasing of an ion cloud oscillating in a disturbed quadratic potential. A method to evaluate the characteristic misphasing time is presented. For the case of disturbance amplitude being constant along ion trajectories, the designated problem may be considered analytically. If the disturbance amplitude depends on oscillation co-ordinate, the result can be obtained by use of numerical integration. An example of numerical calculation is presented. PMID- 18401041 TI - Ion optical evaluation of a miniature double-focusing mass spectrograph. AB - A new miniature double-focusing mass spectrograph has been designed and constructed. The ion optical system was designed based on Mattauch-Herzog geometry. The mass spectrograph employs a focal plane detector consisting of a microchannel plate, a phosphor layer, a fiber-optic plate and a charge-coupled device. For the evaluation of the ion optics of the instrument, the energy and angular focal planes were investigated both experimentally and by simulation. Double focusing was satisfactorily achieved along a straight line over a wide mass range, and the experimental and simulated results were mutually consistent. A second-order element of the transfer matrix was also measured experimentally and proved to be in good agreement with the simulated result. PMID- 18401042 TI - Electron-transfer-induced decomposition of 1,2-dioxetanes in negative-mode matrix assisted laser desorption/ionization time-of-flight mass spectrometry. AB - 1,2-Dioxetanes bearing an aromatic electron donor undergo intramolecular charge transfer-induced chemiluminescence (CTICL). Although there has been some controversy regarding the mechanisms involved, there is little experimental evidence to strongly support any of the proposed mechanisms. In the course of our investigations, to clarify these mechanisms, we tried to effectively ionize dioxetanes bearing a phenolic group and found that poly(3-octylthiophene-2,5 diyl) was a promising matrix for negative-mode matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS). Electron transfer ionization was found to take place for dioxetanes bearing a hydroxyphenyl moiety that had been further substituted with an aromatic group, which acted as an antenna to catch an electron from the matrix. Furthermore, the characteristic fragmentation of dioxetanes 3c-3d was thought to occur by the elimination of 2-methyl-1-propene (56 u) and pivalaldehyde (86 u) from deprotonated ion [M - H](-) of dioxetanes, based on the results of muliple mass spectrometry measurements of dioxetanes using MALDI quadrupole ion trap ToF-MS. Based on a comparison of fragmentation in dioxetanes and the corresponding keto esters, dioxetanes were presumed to initially generate excited keto esters from which fragmentation took place. PMID- 18401043 TI - Differentiation of 2-alkylthioorotic acids, methyl and ethyl 2-alkylthioorotates and hydrazides of 2-alkylthioorotic acids by using electron ionization mass spectra. AB - Electron ionization mass spectra of 12 derivatives of 2-thioorotic acid have been discussed and general fragmentation routes of their molecular ions have been proposed. The compounds under discussion were three groups of four chemical species (2-alkylthioorotic acid, methyl 2-alkylthioorotate, ethyl 2 alkylthioorotate and 2- alkylthioorotic hydrazide) each with the same relative molecular mass. The comparison of selected ions relative abundances and their correlation with the abundance of molecular ions enable differentiation between isomeric or isobaric species in this class of compounds. PMID- 18401044 TI - Structural characterization and differentiation of isomeric omega-bromoalkoxy derivatives of (E)-chalcone by means of mass spectrometry. AB - Mass spectrometry with electron ionization and electrospray ionization have been applied to characterize and differentiate the isomeric ortho-, meta- or para-(E) bromoalkyloxychalcones 1-15. The difference in the values of micro1-micro5 (i.e. the ratio of abundances of the selected fragment ions to those of the molecular ions) in the series of isomeric chalcones studied and the so-called "in-source" fragmentation induced by increasing cone voltage have been found to be important and reliable indicators differentiating the isomers studied. PMID- 18401045 TI - Electrospray tandem mass spectrometry of beta-nitroalkenyl meso tetraphenylporphyrins. AB - Beta-nitroalkenyl meso-tetraphenylporphyrins [beta-TPPCHC(NO(2))R)], as free bases and Zn(II) complexes, were studied by electrospray mass spectrometry (ESI MS). Under this ionisation condition the [M + H](+) ions are formed. The fragmentation pattern of the resulting [M + H](+) ions were studied by electrospray tandem mass spectrometry (ESI-MS/MS). The ESI-MS/MS of beta- nitroalkenylporphyrins, either as free-bases or as Zn(II) complexes, show several interesting features, distinct from the typical behaviour of nitro compounds. For the studied compounds, common main fragmentation patterns are observed, namely characteristic losses of NO(2), HNO(2), 2OH, RNO(2), RCNO, RCNO(2), RCH(2)NO(2), C(6)H(5) plus NO(2) and the formation of the protonated macrocycle, [TPP + H](+) or [ZnTPP + H](+). However, depending on the presence or absence of the metal and the nature of the R substituent, important differences are observed on the relative abundances of the ions formed by the same fragmentation pathway. The presence of bromine in the alkenyl group leads to a peculiar behaviour, since the main fragmentation pattern corresponds to the combined elimination of the bromine atom with the typical nitro group fragments. When R = Br, the loss of the nitro group occurs in low relative abundance (11-16%). However, when R = CH(3), the relative abundance of the ion due to the loss of HNO(2) changes drastically from 100%, observed for the free-base porphyrin, to 29% in the case of the Zn(II) complex. These variations of the relative abundance of the fragment corresponding to the loss of the nitro moiety (typically considered as a diagnostic fragment) can induce to an erroneous interpretation of their MS/MS spectra. Some fragmentations are observed only for the free-base porphyrins, namely the loss of CH(NO(2)R and HNO(2) plus C(2)H(2), while the loss of OH, H(2)O, OH plus H(2)O and RCCH plus H(2)O is observed only for the complexes. Unusual and unexpected fragmentations are also observed, namely the losses of RCNO, RCNO(2) and HNO(2) plus C(2)H(2). This work demonstrates that valuable structural information about the beta-nitroalkenyl substituents linked to meso- tetraarylporphyrins can be achieved using MS/MS. These results can also be useful for the interpretation of the mass spectra of other nitroalkenyl substituted compounds. PMID- 18401046 TI - [Dynamic observation of hormonal characteristics in the newborn babies with periventricular leukomalacia during their first week of lives]. AB - The problem of peculiarity of hormonal adaptation in newborn babies with periventricular leucomalacia--a condition that causes lifelong disability, in spite of its scientific and practical value is not adequately explored. The aim of the research was to investigate hormonal characteristics in the newborn babies with periventricular leucomalacia and to determine clinical and laboratory criteria for early prognosis of periventricular leucomalacia. Dynamic observation was conducted on 48 newborn babies who had acute and traumatic periventricular leucomalacia. Concentration of thyrotrophic hormone (TTH), tiroxine (T4) and triodtironin (T3) were determined in venous blood of the newborn babies on their first and fifth days of life. Cortisol was determined by the radioimmune method (RIA). The investigation revealed definite normal-adaptational reactions. Changes of hormonal traumatic periventricular leucomalacia, manifested in reduction of TTG, T3 level; concentration of T4 was moderately high. PMID- 18401047 TI - [Clinical and immunological indices for prognosis of hypoxic ischemic encephalopathy with periventricular leucomalacia in the newborns]. AB - The aim of the research was to determine clinical and laboratory criteria for early prognosis of Hypoxic-ischemic encephalopathy and periventricular leucomalacia in the newborn babies. 40 newborn babies (68% boys and 32% girls) from their first to thirty days of life were investigated. Results revealed marked immunological disorders which can be used to diagnose diseases. PMID- 18401048 TI - The relationship between endothelial dysfunction and 24-hour blood pressure rhythm in patients with arterial hypertension. AB - The purpose of the study was an investigation of relationships between duration of arterial hypertension (AH), 24-hour blood pressure (BP) rhythm and endothelial function. 18 (mean age 50+/-8.7; 8%-male, 10%-female) outpatients with AH were included in the study. All subjects underwent off-therapy 24-hour ambulatory BP monitoring and vascular Doppler echography of the brachial artery. Hypercholesterolemic subjects, diabetics, smokers and patients with Raynaud's phenomenon were excluded from the study. Compared with dipper women, non-dipper women showed statistically significant lowering of FMD (11.8+/-4.2% vs. 3.4+/ 3.85%, p<0.005). The same fact was observed in men (16.5+/-3.45% dipper vs. 4.0+/ 1.08% non-dipper, p<0.05). Association of the duration of arterial hypertension with FMD was significant (7.9+/-4% (<10 y) vs. 2.0+/-3% (>10 y); p= 0.017). Compared with dipper patients, non-dipper subjects showed an impaired endothelial function (11.4+/-2% vs. 3.5+/-2.6%; p=0.0006). The present data confirm the presence of disturbed endothelium-dependent vasodilatation in arterial hypertension. The study showed significant relation between duration of arterial hypertension and FMD. Furthermore, these results demonstrate that the main factor, which leads to endothelial dysfunction, is the shortage in the lowering BP during the night. PMID- 18401049 TI - [The therapeutic efficiency of complex-ACE inhibitor (perindopril) and traditional antiarrhythmic-treatment for prevention atrial fibrillation paroxysms after successful electric cardio version]. AB - The aim of the study was to estimate the therapeutic efficiency of complex: ACE inhibitor (perindopril) and traditional antiarrhythmic treatment of atrial fibrillation paroxysms after successful electric cardio version for prevention of repeated attacks of atrial fibrillation and for maintenance of normal sinus rhythm in patients. The proposed scheme of treatment reduced number of recurrences, maintained normal sinus rhythm and allowed reaching relatively rapid normalization of mechanical function of left atrium. PMID- 18401050 TI - [Effect of combination therapy with preduktal and prestarium on the left ventricular function in the postinfarction period]. AB - The aim of the study was to estimate the efficiency of combination therapy with Preductal and Prestarium after MI. We investigated 152 patients with acute MI of anterior wall (Q wave) and Heart Failure (II-III class by NYHA). I group included 92 p, which were treated by Preductal (60 mg daily) on the background of the standard therapy. II group (60 p) treated by standard therapy. (ACE inhibitor, diuretic, beta-blocks). We divided the patients in 2 subgroups, with EF>45% and EF<45%. Echo investigation was performed after 1,3,6.12 months from the beginning of MI. We estimated LV EDV and ESV, EF, FS, SV and diastolic function. 12 month after myocardial infarction decreases of average heart failure and angina classes. After 6,12 months was noted decreased EDV and ESV, increased EF, FS,SV in both groups confidence, but spatially in the I gr. Patients, who had EF<45% and were treated with Preductal improved EF with 30,1%(p<0,05) confidence, which was 1,5 times more then in the control gr, also increased FS and SV by 32,7%, 13,6% p<0,05. At the same time was observed diastolic function improvement, particular decreased Ve, Ve/Va, and increased Va specially in the Preductal group more intensive and confidence. The treatment with Preductal during 12 months indicates his positive influence on the LV systolic and diastolic function, which is most significant in the patients with low EF. PMID- 18401051 TI - Bronchial asthma treatment retrospective analysis. AB - Allergology and Clinical Immunology Department of the Tbilisi State Medical University has created and implemented numerous medications and methods in treatment of asthma since its establishment in 1974. The aim of the present study was to analyze the thirty three year experience of the Allergology and Clinical Immunology Department in its attempt to treat bronchial asthma. The retrospective analysis of case histories between 1980 and 2006 period has been carried out to evaluate inpatient treatment efficacy of patients with bronchial asthma in Georgia during different periods of social and economic development. The first group included the period between 1980 and 1984. The second group--between 1996 and 2000 (time of medical reform initiation). The third group--between 2002 and 2006 (state standard implementation and function period). The efficacy of inpatient treatment has been investigated for each of three groups. PMID- 18401052 TI - [Dynamics of hip joint biomechanics in patients with coxarthrosis at the time of hippotherapy]. AB - The problems of degenerative-dystrophic abnormalities stimulate the development of new skills and methods of treatment and rehabilitation of the diseases. The goal of the study was to determine the efficacy of hippotherapy in patients with coxarthrosis, according to functional and biomechanical parameters. Hippotherapy involves the utilization of horseback riding to stimulate the patient's normal reactions and locomotion; to improve the balance and coordination of movement, normalize muscle tension, and eliminate pathological reflexes. The advantage of the hippotherapy is in the specific posture, which is adopted by hip joint at the time of riding and in movement, which is accomplished by rider, at different paces of the horse. 10 female patients from 14 to 32 years old with coxarthrosis were under the observation. The rehabilitation of the patients was carried out by means of hippotherapy, which consisted of three months riding three times a week. To evaluate the efficacy of treatment, a new method of biomechanical registration of hip joint movement during hippotherapy on pacing horse was developed. The dynamics of biomechanical curves before and after the treatment, as well as the clinical and functional parameters of the patients allowed the authors to conclude: hippotherapy improves a hip joint functional state in patients with coxarthrosis; improves the muscle-tendineous component of hip joint movement. Hippotherapy may be considered as the pathogenetic method of treatment of coxarthrosis. Drawing the biomechanical curve of hip joint movement at the time of riding is the objective method of studying its function. PMID- 18401053 TI - [Epidemiological and structural features of prenatal and perinatal risk factors involved in central nervous system injuries]. AB - Different prenatal and perinatal factors are the main reason of psycho-motor development delay. Early reveal of risks factors determine in newborns is the guarantor of the maximal decrease in physical and mental inability. The goal of the study was to investigate the epidemiological and structural features of the neurological diseases and to reveal the risk factors for neurological diseases in newborns. The results of the investigation of 1272 newborns are reported. In the early neonatal period the newborns were divided into three groups: practically healthy newborns, newborns with pathological diseases. The group of risk included newborns with the risk of development of pathologies because of the certain prenatal and perinatal factors. Risk factors which may be the reason of a delay of intrauterine development of the child or/ and the damage of central nervous system. Results of investigation coincide with the reference data, according to which the different prenatal and perinatal risks factors promote the development of different neurological diseases of newborn in future. PMID- 18401054 TI - [Repeated general anesthesia, as a component of intensive therapy of critical conditions in children with severe burns]. AB - Adequate maintenance of anaesthesia during bandaging of burn injuries in children remains to be the problem of current importance. The authors analyzed 166 bandage changes under anesthetic support (children under ketamine with various types of premedication). It is revealed, that the ketamine anesthesia irrespective of a way of its introduction stimulates cardiovascular system and reduces parameters of external breath. With an intramuscular dose of 10 mg/kg of body weight, or intravenous dose of 3 mg/kg as employed in anaesthesia, the effect of ketamine continues for 12 minutes. It is found that intravenous dose of 3 mg/kg ketamine, has the same effect on homodynamic and breath as the dose of 5 mg/kg ketamine. The intravenous infusion of 5 mg/kg ketamine continued anesthesia for 17 minutes. Premedication of diazepam, extended for 4 minutes duration of anesthesia. It is concluded that ketamine-diazepam combinations can be very useful for induction of general anaesthesia. PMID- 18401055 TI - [Ophthalmological problems in children as distant earthquake consequences in Armenia]. AB - The special program was introduced by HKCC Govard Karagezian in 2000-2002 to provide eye examinations for children in zone of earthquake (Gyumri and Vanadzor) in Armenia. Specialized regional services were established. The aim of the program was the improvement of pediatric health care and accessibility of modern ophthalmologic services, and investigation of distant consequences of earthquake. The screening revealed that 3696 of 24.877 Gyumri suffer from weak vision (148,6+/-2,3). Weak vision was found in 2407 of 13.005 children from 7 to 14 years old in Vanadzor (185,1+/-3,4). 2. 443 children in Gyumri and 2.593 children in Vanadzor had eye pathology. The socio-economic consequences of Spitak earthquake were also investigated. 52,9% of children suffering from eye problems live or study in temporary shelters (metal or wooden) in Gyumri and in Vanadzor- 4,97%. The refractive errors are prevalent among schoolchildren--62,1% in Gyumri and 55,5% in Vanadzor. PMID- 18401056 TI - The mechanisms of neurodegenerative processes and current pharmacotherapy of Alzheimer's disease. AB - An increasing number of studies suggest that current pharmacotherapy used in Alzheimer's disease (AD), in addition to having a symptomatic effect, also may interact with the ongoing neuropathological processes in the brain. The oldest hypothesis explain the cause of the AD is the "cholinergic hypothesis", which states, that AD begins as a deficiency in the production of acetylcholine (ACh). Interactions between the cholinergic and serotoninergic systems are believed to play a role in the mechanism underlying AD. The activation of NMDA receptors and increase in intracellular Ca(++) concentration play key role in the development of neurodegenerative processes. Potassium channels may also be involved in several other steps within the cascade that leads to neurodegeneration. In the development of AD a great role play an imbalance between anabolism and catabolism causes an accumulation of amyloid beta-peptide (Abeta), which is a proposed trigger of the onset of AD. There are various therapeutic strategies in current pharmacotherapy of AD. Major group is inhibitors of acetylcholinesterase- donepezil, galantamine, physostigmine. The new effective treatments of AD are NMDA glutamate receptor antagonist-memantine and potassium channel openers such as retigabine. Experimental study suggest, that neprilisin, a rate-limiting peptidase, decreases neurodegeneration. PMID- 18401057 TI - Anti-tumor preventative effect of mono-therapy with the use of proteus vaccine, Staphylococcus antitoxin and divaccine of Staphylococcus-proteus. AB - Anti-tumor preventative effect of mono-therapy with the use of Proteus vaccine, Staphylococcus antitoxin and divaccine of Staphylococcus-Proteus has been studied. Experiments were carried out on 80 non-purebred laboratory white mice (age--3-3,5 months, body mass--18-20 g) and 60 rats (body mass--100-120 g) using intraperitoneal inoculation of Ehrlich's adenocarcinoma (ascitic form--EAT, in mice, cancer cells--3 x 10(6)), and subcutaneous inoculation of Sarcoma S-45 (in rats). Anti-tumor preventative effect of bacterial vaccines and immunization was evaluated according to the following parameters: Frequency of cancer development, Inhibition of cancer growth, Body mass index of experimental animals, Volume of ascitic fluid. Results of experiments have shown that use of bacterial polysaccharides with preventative purposes has better effect at S-45 growth than at EAT growth; Vaccination with Proteus prolongs lifespan mach more than vaccination with antitoxin of Staphylococcus; Vaccination with complex divaccine of Staphylococcus-Proteus causes complete resorption of tumors from 32 to 60 days; Development of experimental malignant tumors depends on type of anti microbial vaccines and starting date of inoculation after completion of vaccination. PMID- 18401058 TI - [Factors associated with the health status of Georgian dance studio students]. AB - The article investigates factors associated with occupational disease among Georgian dance studio students. The problem of work-related neuromuscular and musculoskeletal disorders is outlined. Reiterating and stereotypical movements cause tension of specific muscles, constrained body, monotonous work, and emotional tension lead to occupational support-motor apparatus disorders. The aim of the work is to study the health status of Georgian dance studio students. 254 Georgian dance studio students were examined. It was found that pathology of cardio-vascular, respiratory and digesting systems was more frequent among male students than females; musculoskeletal disorders were common only among the girls. 1/3 of girls with disorder of musculoskeletal system had inborn dysplasia of hip joint bones. In three male students hernia had developed during the first year of the study. PMID- 18401059 TI - Assessment of the sources of error affecting the quantitative accuracy of SPECT imaging in small animals. AB - Small animal SPECT imaging systems have multiple potential applications in biomedical research. Whereas SPECT data are commonly interpreted qualitatively in a clinical setting, the ability to accurately quantify measurements will increase the utility of the SPECT data for laboratory measurements involving small animals. In this work, we assess the effect of photon attenuation, scatter and partial volume errors on the quantitative accuracy of small animal SPECT measurements, first with Monte Carlo simulation and then confirmed with experimental measurements. The simulations modeled the imaging geometry of a commercially available small animal SPECT system. We simulated the imaging of a radioactive source within a cylinder of water, and reconstructed the projection data using iterative reconstruction algorithms. The size of the source and the size of the surrounding cylinder were varied to evaluate the effects of photon attenuation and scatter on quantitative accuracy. We found that photon attenuation can reduce the measured concentration of radioactivity in a volume of interest in the center of a rat-sized cylinder of water by up to 50% when imaging with iodine-125, and up to 25% when imaging with technetium-99m. When imaging with iodine-125, the scatter-to-primary ratio can reach up to approximately 30%, and can cause overestimation of the radioactivity concentration when reconstructing data with attenuation correction. We varied the size of the source to evaluate partial volume errors, which we found to be a strong function of the size of the volume of interest and the spatial resolution. These errors can result in large (>50%) changes in the measured amount of radioactivity. The simulation results were compared with and found to agree with experimental measurements. The inclusion of attenuation correction in the reconstruction algorithm improved quantitative accuracy. We also found that an improvement of the spatial resolution through the use of resolution recovery techniques (i.e. modeling the finite collimator spatial resolution in iterative reconstruction algorithms) can significantly reduce the partial volume errors. PMID- 18401060 TI - Non-uniform depth scanning for proton therapy systems employing active energy variation. AB - In proton scanning systems that employ active energy variation for depth modulation, a switch of the particle energy might typically require 1-2 s. For plans comprising many energy slices, these seconds could sum up to a non negligible fraction of the total treatment duration. We have applied the Nyquist Shannon sampling theorem to determine an efficient spatial arrangement of Bragg peaks in a target volume. This pre-determined schedule of increasing energy spacing with higher energy allows us to reduce the number of used energy slices without compromising the physical dosimetric quality of a plan. Our results suggest that the advantage of such a simple implementation would be especially significant for larger, deep-seated tumors such as the prostate; the number of energy slices was cut by a factor of 2-6. PMID- 18401061 TI - The development and investigation of a prototype three-dimensional compensator for whole brain radiation therapy. AB - Whole brain radiation therapy (WBRT) is the standard treatment for patients with brain metastases, and is often used in conjunction with stereotactic radiotherapy for patients with a limited number of brain metastases, as well as prophylactic cranial irradiation. The use of open fields (conventionally used for WBRT) leads to higher doses to the brain periphery if dose is prescribed to the brain center at the largest lateral radius. These dose variations potentially compromise treatment efficacy and translate to increased side effects. The goal of this research was to design and construct a 3D 'brain wedge' to compensate dose heterogeneities in WBRT. Radiation transport theory was invoked to calculate the desired shape of a wedge to achieve a uniform dose distribution at the sagittal plane for an ellipsoid irradiated medium. The calculations yielded a smooth 3D wedge design to account for the missing tissue at the peripheral areas of the brain. A wedge was machined based on the calculation results. Three ellipsoid phantoms, spanning the mean and +/- two standard deviations from the mean cranial dimensions were constructed, representing 95% of the adult population. Film was placed at the sagittal plane for each of the three phantoms and irradiated with 6 MV photons, with the wedge in place. Sagittal plane isodose plots for the three phantoms demonstrated the feasibility of this wedge to create a homogeneous distribution with similar results observed for the three phantom sizes, indicating that a single wedge may be sufficient to cover 95% of the adult population. The sagittal dose is a reasonable estimate of the off-axis dose for whole brain radiation therapy. Comparing the dose with and without the wedge the average minimum dose was higher (90% versus 86%), the maximum dose was lower (107% versus 113%) and the dose variation was lower (one standard deviation 2.7% versus 4.6%). In summary, a simple and effective 3D wedge for whole brain radiotherapy has been developed. The wedge gives a more uniform dose distribution than commonly used techniques. Further development and shape optimization may be necessary prior to clinical implementation. PMID- 18401062 TI - Calculated SAR distributions in a human voxel phantom due to the reflection of electromagnetic fields from a ground plane between 65 MHz and 2 GHz. AB - If an electromagnetic field is incident normally onto a perfectly conducting ground plane, the field is reflected back into the domain. This produces a standing wave above the ground plane. If a person is present within the domain, absorption of the field in the body may cause problems regarding compliance with electromagnetic guidelines. To investigate this, the whole-body averaged specific energy absorption rate (SAR), localised SAR and ankle currents in the voxel model NORMAN have been calculated for a variety of these exposures under grounded conditions. The results were normalised to the spatially averaged field, a technique used to determine a mean value for comparison with guidelines when the field varies along the height of the body. Additionally, the external field values required to produce basic restrictions for whole-body averaged SAR have been calculated. It was found that in all configurations studied, the ICNIRP reference levels and IEEE MPEs provided a conservative estimate of these restrictions. PMID- 18401063 TI - Quantification of interplay effects of scanned particle beams and moving targets. AB - Scanned particle beams and target motion interfere. This interplay leads to deterioration of the dose distribution. Experiments and a treatment planning study were performed to investigate interplay. Experiments were performed with moving radiographic films for different motion parameters. Resulting dose distributions were analyzed for homogeneity and dose coverage. The treatment planning study was based on the time-resolved computed tomography (4DCT) data of five lung tumor patients. Treatment plans with margins to account for respiratory motion were optimized, and resulting dose distributions for 108 different motion parameters for each patient were calculated. Data analysis for a single fraction was based on dose-volume histograms and the volume covered with 95% of the planned dose. Interplay deteriorated dose conformity and homogeneity (1-standard deviation/mean) in the experiments as well as in the treatment-planning study. The homogeneity on radiographic films was below approximately 80% for motion amplitudes of approximately 15 mm. For the treatment-planning study based on patient data, the target volume receiving at least 95% of the prescribed dose was on average (standard deviation) 71.0% (14.2%). Interplay of scanned particle beams and moving targets has severe impact on the resulting dose distributions. Fractionated treatment delivery potentially mitigates at least parts of these interplay effects. However, especially for small fraction numbers, e.g. hypo fractionation, treatment of moving targets with scanned particle beams requires motion mitigation techniques such as rescanning, gating, or tracking. PMID- 18401064 TI - Quantification of cardiac magnetic field orientation during ventricular de- and repolarization. AB - We compared the stability and discriminatory power of different methods of determining cardiac magnetic field map (MFM) orientation within the context of coronary artery disease (CAD). In 27 healthy subjects and 24 CAD patients, multichannel magnetocardiograms were registered at rest. MFM orientation was determined during QT interval using: (a) locations of the positive and negative centres-of-gravity, (b) locations of the field extrema and (c) the direction of the maximum field gradient. Deviation from normal orientation quantified the ability of each approach to discriminate between healthy and CAD subjects. Although the course of orientation was similar for all methods, receiver operating characteristic analysis showed the best discrimination of CAD patients for the centre-of-gravity approach (area-under-the-curve = 86%), followed by the gradient (84%) and extrema (76%) methods. Consideration of methodological and discriminatory advantages with respect to noninvasive diagnosis of CAD suggests that the centres-of-gravity method is the most suited one. PMID- 18401065 TI - Real-time monitoring and diagnosis of scintillation dosimeters using an ultraviolet light emitting diode. AB - Plastic scintillator fibre optic dosimeters (FODs) have advantages for both brachytherapy and external beam radiotherapy applications. Convenient real-time monitoring and diagnosis of such dosimeters are desirable because of changes in the optical circuit that may arise in use. In this paper, we propose and demonstrate a real-time method using ultraviolet light emitting diodes (LED) to stimulate the scintillator and to diagnose failures of FODs. Key aspects of the LED FOD dosimetry design are investigated, enabling the design of a stable and accurate real-time monitoring dosimetry system. We demonstrate experimentally that the real-time monitoring FOD system is convenient to be used to monitor FOD dosimeters and to diagnose their failures resulted from different mechanisms. PMID- 18401066 TI - Speed optimized influence matrix processing in inverse treatment planning tools. AB - An optimal plan in modern treatment planning tools is found through the use of an iterative optimization algorithm, which deals with a high amount of patient related data and number of treatment parameters to be optimized. Thus, calculating a good plan is a very time-consuming process which limits the application for patients in clinics and for research activities aiming for more accuracy. A common technique to handle the vast amount of radiation dose data is the concept of the influence matrix (DIJ), which stores the dose contribution of each bixel to the patient in the main memory of the computer. This study revealed that a bottleneck for the optimization time arises from the data transfer of the dose data between the memory and the CPU. In this note, we introduce a new method which speeds up the data transportation from stored dose data to the CPU. As an example we used the DIJ approach as is implemented in our treatment planning tool KonRad, developed at the German Cancer Research Center (DKFZ) in Heidelberg. A data cycle reordering method is proposed to take the advantage of modern memory hardware. This induces a minimal eviction policy which results in a memory behaviour exhibiting a 2.6 times faster algorithm compared to the naive implementation. Although our method is described for the DIJ approach implemented in KonRad, we believe that any other planning tool which uses a similar approach to store the dose data will also benefit from the described methods. PMID- 18401067 TI - Tiny a priori knowledge solves the interior problem in computed tomography. AB - Based on the concept of differentiated backprojection (DBP) (Noo et al 2004 Phys. Med. Biol. 49 3903, Pan et al 2005 Med. Phys. 32 673, Defrise et al 2006 Inverse Problems 22 1037), this paper shows that the solution to the interior problem in computed tomography is unique if a tiny a priori knowledge on the object f(x, y) is available in the form that f(x, y) is known on a small region located inside the region of interest. Furthermore, we advance the uniqueness result to obtain more general uniqueness results which can be applied to a wider class of imaging configurations. We also develop a reconstruction algorithm which can be considered an extension of the DBP-POCS (projection onto convex sets) method described by Defrise et al (2006 Inverse Problems 22 1037), where we not only extend this method to the interior problem but also introduce a new POCS algorithm to reduce computational cost. Finally, we present experimental results which show evidence that the inversion corresponding to each obtained uniqueness result is stable. PMID- 18401068 TI - The physics, biophysics and technology of photodynamic therapy. AB - Photodynamic therapy (PDT) uses light-activated drugs to treat diseases ranging from cancer to age-related macular degeneration and antibiotic-resistant infections. This paper reviews the current status of PDT with an emphasis on the contributions of physics, biophysics and technology, and the challenges remaining in the optimization and adoption of this treatment modality. A theme of the review is the complexity of PDT dosimetry due to the dynamic nature of the three essential components -- light, photosensitizer and oxygen. Considerable progress has been made in understanding the problem and in developing instruments to measure all three, so that optimization of individual PDT treatments is becoming a feasible target. The final section of the review introduces some new frontiers of research including low dose rate (metronomic) PDT, two-photon PDT, activatable PDT molecular beacons and nanoparticle-based PDT. PMID- 18401069 TI - Reproducibility of infrared thermography measurements in healthy individuals. AB - The aim of this study was to investigate the reproducibility of skin surface infrared thermography (IRT) measurements and determine the factors influencing the variability of the measured values. While IRT has been widely utilized in different clinical conditions, there are few available data on the values of the skin temperature patterns of healthy subjects and their reproducibility. We recorded the whole body skin temperatures of sixteen healthy young men with two observers on two consecutive days. The results were compared using intra-class correlations analyses (ICC). The inter-examiner reproducibility of the IRT measurements was high: mean ICC 0.88 (0.73-0.99). The day-to-day stability of thermal patterns varied depending on the measured area: it was high in the core and poor in distal areas. The reproducibility of the side-to-side temperature differences (deltaT) was moderately good between the two observers (mean ICC 0.68) but it was reduced with time, especially in the extremities, mean ICC 0.4 ( 0.01-0.83). The results suggest that the IRT technique may represent an objective quantifiable indicator of autonomic disturbances although there are considerable temporal variations in the measured values which are due to both technical factors such as equipment accuracy, measurement environment and technique, and physiological variability of the blood flow, and these factors should be taken into account. PMID- 18401070 TI - Continuous estimates of dynamic cerebral autoregulation: influence of non invasive arterial blood pressure measurements. AB - Temporal variability of parameters which describe dynamic cerebral autoregulation (CA), usually quantified by the short-term relationship between arterial blood pressure (BP) and cerebral blood flow velocity (CBFV), could result from continuous adjustments in physiological regulatory mechanisms or could be the result of artefacts in methods of measurement, such as the use of non-invasive measurements of BP in the finger. In 27 subjects (61+/-11 years old) undergoing coronary artery angioplasty, BP was continuously recorded at rest with the Finapres device and in the ascending aorta (Millar catheter, BP(AO)), together with bilateral transcranial Doppler ultrasound in the middle cerebral artery, surface ECG and transcutaneous CO(2). Dynamic CA was expressed by the autoregulation index (ARI), ranging from 0 (absence of CA) to 9 (best CA). Time varying, continuous estimates of ARI (ARI(t)) were obtained with an autoregressive moving-average (ARMA) model applied to a 60 s sliding data window. No significant differences were observed in the accuracy and precision of ARI(t) between estimates derived from the Finapres and BP(AO). Highly significant correlations were obtained between ARI(t) estimates from the right and left middle cerebral artery (MCA) (Finapres r=0.60+/-0.20; BP(AO) r=0.56+/-0.22) and also between the ARI(t) estimates from the Finapres and BP(AO) (right MCA r=0.70+/-0.22; left MCA r=0.74+/-0.22). Surrogate data showed that ARI(t) was highly sensitive to the presence of noise in the CBFV signal, with both the bias and dispersion of estimates increasing for lower values of ARI(t). This effect could explain the sudden drops of ARI(t) to zero as reported previously. Simulated sudden changes in ARI(t) can be detected by the Finapres, but the bias and variability of estimates also increase for lower values of ARI. In summary, the Finapres does not distort time-varying estimates of dynamic CA obtained with a sliding window combined with an ARMA model, but further research is needed to confirm these findings in healthy subjects and to assess the influence of different physiological manoeuvres. PMID- 18401071 TI - Study of the motion artefacts of skin-mounted inertial sensors under different attachment conditions. AB - A common problem shared by accelerometers, inertial sensors and any motion measurement method based on skin-mounted sensors is the movement of the soft tissues covering the bones. The aim of this work is to propose a method for the validation of the attachment of skin-mounted sensors. A second-order (mass-spring damper) model was proposed to characterize the behaviour of the soft tissue between the bone and the sensor. Three sets of experiments were performed. In the first one, different procedures to excite the system were evaluated to select an adequate excitation stimulus. In the second one, the selected stimulus was applied under varying attachment conditions while the third experiment was used to test the model. The heel drop was chosen as the excitation method because it showed lower variability and could discriminate between different attachment conditions. There was, in agreement with the model, a trend to increase the natural frequency of the system with decreasing accelerometer mass. An important result is the development of a standard procedure to test the bandwidth of skin mounted inertial sensors, such as accelerometers mounted on the skin or markers heavier than a few grams. PMID- 18401072 TI - The role of the autonomic nervous system in hypertension: a bond graph model study. AB - A bond graph model of the cardiovascular system with embedded autonomic nervous regulation was developed for a better understanding of the role of the autonomic nervous system (ANS) in hypertension. The model is described by a pump model of the heart and a detailed representation of the head and neck, pulmonary, coronary, abdomen and extremity circulation. It responds to sympathetic and parasympathetic activities by modifying systemic peripheral vascular resistance, heart rate, ventricular end-systolic elastance and venous unstressed volumes. The impairment of ANS is represented by an elevation of the baroreflex set point. The simulation results show that, compared with normotensive, in hypertension the systolic and diastolic blood pressure (SBP/DBP) rose from 112/77 mmHg to 144/94 mmHg and the left ventricular wall thickness (LVWT) increased from 10 mm to 12.74 mm. In the case that ANS regulation was absent, both the SBP and DBP further increased by 8 mmHg and the LVWT increased to 13.22 mm. The results also demonstrate that when ANS regulation is not severely damaged, e.g. the baroreflex set point is 97 mmHg, it still has an effect in preventing the rapid rise of blood pressure in hypertension; however, with the worsening of ANS regulation, its protective role weakens. The results agree with human physiological and pathological features in hemodynamic parameters and carotid baroreflex function curves, and indicate the role of ANS in blood pressure regulation and heart protection. In conclusion, the present model may provide a valid approach to study the pathophysiological conditions of the cardiovascular system and the mechanism of ANS regulation. PMID- 18401073 TI - The hydraulic conductivity of Matrigel. AB - In this study, we measured the specific hydraulic conductivity (K) of Matrigel at 1% and 2% concentrations as a function of perfusion pressure (0 to 100 mmHg) and compared the results to predictions from two models: a fiber matrix model that predicted K of the gel based upon its composition, and a biphasic model that predicted changes in K caused by pressure induced compaction of the gels. The extent of gel compaction as a function of perfusion pressure was also assessed, allowing us to estimate the stiffness of the gels. As expected, 2% Matrigel had a lower K and a higher stiffness than did 1% Matrigel. Measured values of K of both 1% and 2% Matrigel samples showed good agreement with the predictions of the fiber matrix model. Pressure-induced changes in K were better described by the biphasic model than a model in which uniform compression of the gel was assumed. We conclude that K of multi-component gels, such as Matrigel can be well characterized by fiber matrix models, and that pressure-induced changes in K of such gels can be well characterized by biphasic models. PMID- 18401074 TI - Mechanical response of a living human epidermal keratinocyte sheet as measured in a composite diaphragm inflation experiment. AB - Sheets of normal human epidermal keratinocytes (NHEKs) were reconstituted in vitro on tensed but highly compliant, freestanding polydimethylsiloxane (PDMS) membranes, 5.0 mm in diameter and 10 mum thick. NHEK-PDMS composite diaphragm (CD) specimens were then subjected to cyclical axisymmetric inflation tests to investigate epithelial sheet rheology under conditions of physiologically severe deformations (~50% nominal polar biaxial strains). Because the compliance of the specially formulated PDMS membrane was greater than that of the attached cell layer, the finite load-deformation behavior (mechanical response) of the living NHEK sheet was inferred from differences between the mechanical behavior of the CD specimen and the response of the underlying PDMS membrane measured prior to cell culture. In these composite diaphragm inflation (CDI) experiments, interconnected NHEKs exhibited rheological behaviors that were suggestive of a viscoelastic-plastic stress response. Remarkably, specimens returned to quiescent culture following a sequence of inflation tests recovered at least 80% of their original ability to store elastic strain energy, evidence of biological adaptation and recovery or restitutio ad integrum. Unlike methodologies that assay the morphological or biochemical response of cultured cells to an applied mechanostimulus, CDI experiments can be used to probe the load-bearing functions of desmosomes and adherens junctions within a living epithelial sheet, as well as to assess the rheological behaviors of the intermediate filament and microfilament networks that these cell-cell junctions serve to interconnect. PMID- 18401075 TI - Effect of lanthanum on red blood cell deformability. AB - Prior reports describing the effects of lanthanum (La(3+)) on red blood cells (RBC) have focused on the effects of this lanthanide on cell fusion or on membrane characteristics (e.g., ion movement across membrane, membrane protein aggregation); the present study explores its rheological and biophysical effects. Normal human RBC were exposed to La(3+) levels up to 200 microM then tested for: (1) cellular deformability using a laser-based ektacytometer and an optical-based rheoscope; (2) membrane viscoelastic behavior via micropipettes; (3) surface charge via micro electrophoresis. La(3+) concentrations of 12.5 to 200 microM caused dose-dependent decreases of deformability that were greatest at low stresses: these rheological changes were completely reversible upon removing La(3+) from the media either by washing with La(3+)-free buffer or by suspending La(3+)-exposed cells in La(3+)-free media (i.e., viscous dextran solution). Both membrane shear elastic modulus and membrane surface viscosity were increased by 25-30% at 100 or 200 microM. As expected, La(3+) decreased RBC electrophoretic mobility (EPM), with EPM inversely but not linearly associated with deformability; changes of EPM were also completely reversible. These results thus indicate novel aspects of RBC cellular and membrane rheological behavior yet raise questions regarding specific mechanisms responsible for La(3+)-induced alterations. PMID- 18401076 TI - Volume flow and wall shear stress quantification in the human conjunctival capillaries and post-capillary venules in vivo. AB - Understanding the mathematical relationships of volume blood flow and wall shear stress with respect to microvessel diameter is necessary for the study of vascular design. Here, for the first time, volume flow and wall shear stress were quantified from axial red blood cell velocity measurements in 104 conjunctival microvessels of 17 normal human volunteers. Measurements were taken with a slit lamp based imaging system from the post capillary side of the bulbar conjunctiva in microvessel diameters ranging from 4 to 24 micrometers. The variation of the velocity profile with diameter was taken into account by using a profile factor function. Volume flow ranged from 5 to 462 pl/s with a mean value of 102 pl/s and gave a second power law best fitting line (r=0.97) deviating significantly from the third power law relation with diameter. The estimated wall shear stress declined hyperbolically (r=0.93) from a maximum of 9.55 N/m(2) at the smallest capillaries, down to a minimum of 0.28 N/m(2) at the higher diameter post capillary venules. The mean wall shear stress value for all microvessels was 1.54 N/m(2). PMID- 18401077 TI - Viscoelastic properties of an exopolysaccharide: Aeromonas gum, produced by Aeromonas nichidenii 5797. AB - The viscoelastic properties of Aeromonas (A) gum in water were investigated by using the Rheometric Scientific ARES controlled strain rheometer. An intrinsic viscosity of 8336 ml/g was obtained according to the Fuoss-Straus equation. The effect of salt concentration on intrinsic viscosity revealed that the A gum exists as semiflexible chain. Typical shear-thinning (pseudoplastic) behavior was observed at concentrations higher than 0.52%. The zero shear viscosity (eta(0)) increased with increasing polysaccharide concentration (c) showing a gradient of approximately 1.0, 2.9 and 4.8 in different concentration domains. The critical concentrations c* and c**, at which the transitions from a dilute solution of independently moving chains to semidilute and then concentrated domains occurred, were determined roughly to be 1.2% and 3.5%. The results from dynamic experiments revealed that the A gum solution shows characteristics of polymer solutions without any evidence of gel-like character. All the results from steady and dynamic tests suggest that the A gum is a non-gelling polysaccharide. The temperature dependence of apparent viscosity was described by Arrhenius equation and the flow activation energy was estimated to be 45.2 kJ/mol, which is independent on polymer concentration. PMID- 18401078 TI - High frequency viscoelastic behaviour of low molecular weight hyaluronic acid water solutions. AB - Hyaluronic acid (HA) is a polysaccharide widely used in biomedical applications, due to its elevated biocompatibility and the peculiar viscoelastic properties of its solutions. Although the viscoelastic behaviour of HA solutions has been extensively studied in the literature it has been often reported in the range of low frequency (1-100 Hz) and high salt concentration, whereas the main rheological peculiarities of this molecule are expected at high frequency (>100 Hz) and low salt concentration. In this work we studied the viscoelastic properties of low molecular weight HA (155 kDa) in wide range of concentrations (0.01-20 mg/ml) at low ionic strength and over an extended frequency range (0.1 1000 Hz) using both optical tweezers and conventional rheometry. Good agreement between the high frequency dynamic behaviour (optical tweezers) and the viscoelastic properties at low frequency (rheometry) was found. We also found that, in apparent contradiction with polyelectrolyte solution theory, HA solution behaves as liquid-like viscoelastic fluid (G''>G') even at concentrations higher than the entanglement concentration where a weak-gel behavior should be expected. PMID- 18401079 TI - Concepts on the pathogenesis of adolescent idiopathic scoliosis. Bone growth and mass, vertebral column, spinal cord, brain, skull, extra-spinal left-right skeletal length asymmetries, disproportions and molecular pathogenesis. AB - There is no generally accepted scientific theory for the causes of adolescent idiopathic scoliosis (AIS). Encouraging advances thought to be related to AIS pathogenesis have recently been made in several fields including anthropometry of bone growth, bone mass, spinal growth modulation, extra-spinal left-right skeletal length asymmetries and disproportions, magnetic resonance imaging of vertebral column, spinal cord, brain, skull, and molecular pathogenesis. These advances are leading to the evaluation of new treatments including attempts at minimally invasive surgery on the spine and peri-apical ribs. Several concepts of AIS are outlined indicating their clinical applications but not their research potential. The concepts, by derivation morphological, molecular and mathematical, are addressed in 15 sections: 1) initiating and progressive factors; 2) relative anterior spinal overgrowth; 3) dorsal shear forces that create axial rotational instability; 4) rotational preconstraint; 5) uncoupled, or asynchronous, spinal neuro-osseous growth; 6) brain, nervous system and skull; 7) a novel neuro osseous escalator concept based on a putative abnormality of two normal polarized processes namely, a) increasing skeletal dimensions, and b) the CNS body schema - both contained within a neuro-osseous timing of maturation (NOTOM) concept; 8) transverse plane pelvic rotation, skeletal asymmetries and developmental theory; 9) thoraco-spinal concept; 10) origin in contracture at the hips; 11) osteopenia; 12) melatonin deficiency; 13) systemic melatonin-signaling pathway dysfunction; 14) platelet calmodulin dysfunction; and 15) biomechanical spinal growth modulation. From these concepts, a collective model for AIS pathogenesis is formulated. The central concept of this model includes the body schema of the neural systems, widely-studied in adults, that control normal posture and coordinated movements with frames of reference in the posterior parietal cortex. The escalator concept has implications for the normal development of upright posture, and the evolution in humans of neural control, the trunk and unique bipedal gait. PMID- 18401080 TI - Idiopathic scoliosis and chaos. AB - At growing ages, the progression of the idiopathic scoliosis with a curve under 25 degrees outlines many features related to the chaos theory. The image of the scoliosis calls to mind the "strange attractors" of the chaotic spine. We describe the 7 main characteristics of the dynamical scoliotic system classified as chaotic. It is an open set system, unpredictable, multi-factorial complex, discontinuous with thresholds that you can model, and it is an inter-phase between childhood and adult time. The chaotic model enables us to understand more the progression of the idiopathic scoliosis. It positively modifies the speech with the patient and its family as well as the therapeutic treatment. PMID- 18401081 TI - How can we achieve success in understanding the aetiology of AIS? AB - A cure to prevent scoliosis from developing does not seem to be available in the near future. Primarily this is because of a lack of understanding of the aetiology of this devastating disease or cosmetic deformity. While extensive research has been performed in this area over the past 100 years many experiments have been poorly designed because they have been developed on the premise that patients with AIS all have the same, single underlying cause despite much evidence to the contrary. Consequently, much of the data in the literature can be challenged and perhaps explains the lack of significant progress. Certainly, the results from this previous research suggest strongly that a new approach needs to be adopted or the same confusing results will continue to be collected and little progress will be made. There are certain areas of research that hold the greatest potential for success in finding a cure. These are identified in this paper and included in a theoretical research laboratory. It is suggested that this laboratory need not be theoretical if modern, cheap communication systems were readily adopted throughout the world and if people were willing to share ideas readily and contact each other regularly. In perhaps an unconventional way, the emphasis of this paper is on finding a cure to prevent scoliosis from developing and uses the area of research into the aetiology of scoliosis as the platform for discussion. PMID- 18401082 TI - Mechanical modulation of spinal growth and progression of adolescent scoliosis. AB - It is unclear why some children with a small magnitude scoliosis at the onset of the adolescent growth spurt develop a progressive curve. Normally the skeleton grows symmetrically, presumably because genetic and epigenetic factors regulating growth to maintain growth symmetry despite activities and environmental factors causing asymmetrical loading of the spine. This chapter reviews the recently published data relating to the notion that progression of scoliosis is a result of biomechanical factors modulating spinal growth ('vicious cycle' theory). Quantitative data exist for the key variables in an analysis of scoliosis curve progression. In a predictive model of the evolution of scoliosis simulating the 'vicious cycle' theory, and using these published data, a small lateral curvature of the spine can produce asymmetrical spinal loading that causes asymmetrical growth and a self-perpetuating progressive deformity during skeletal growth. This can occur if the neuromuscular control of muscle activation is directed at minimizing the muscular stress (force per unit cross section), although other activation strategies may produce differing spinal growth patterns. Mechanical modulation of vertebral growth is a significant contributor to the progression of an established scoliosis deformity. Quantitative simulation of this mechanism demonstrates how therapeutic interventions to alter neuromuscular control of trunk muscles or otherwise modify spinal loading may alter the natural history of progression. PMID- 18401083 TI - School screening as a research tool in epidemiology, natural history and aetiology of idiopathic scoliosis. AB - The aim of school screening is to identify most or all the individuals with unrecognized idiopathic scoliosis (IS) at an early stage when a less invasive treatment is more effective. However like other medical screening programs it has not escaped controversy about its value. The present study summarises the contribution of school screening in research of IS epidemiology, natural history and aetiology. Such contribution is beyond the original aim of school screening but is very important to expand our knowledge and adequately understand the pathogenesis of IS. The role of biological factors such as the menarche, the lateralization of the brain, the handedness, the thoracic cage, the intervertebral disc, the melatonin secretion, as well as the role of environmental factors such as the light and the impact of the geographical latitude in IS prevalence were studied in children referred from school screening. The present study provides evidence to support that school screening programs should be continued not only for early detection of IS but also as a basis for epidemiological surveys until we learn much more about the aetiology of IS. PMID- 18401084 TI - Scoliosis and the human genome project. AB - Understanding the cause of a disease or disorder is key to developing effective and humane strategies for early intervention and treatment. School screening programs have made it possible to demonstrate the high prevalence of childhood scoliosis, worldwide, and to reliably identify spinal curvatures early in the disease process before progression to a fixed structural deformity. Unfortunately, effective early interventions have not been established. Developing strategies to prevent scoliosis has been compromised, in general, by lack of understanding of its causes on a case by case basis. Information about genetic loci associated with disorders including scoliosis is emerging rapidly, since completion of the human genome sequence in 2003. These data can be used to identify children at high risk for developing spinal deformities and to design strategies for prevention. PMID- 18401085 TI - How to improve the effectiveness of school screening for idiopathic scoliosis. AB - The value of school screening for idiopathic scoliosis (IS) has been questioned recently, because of its high false positive referrals and its excessive cost, although in areas where screening programs exist, fewer patients ultimately require surgery for IS. In a typical school screening setting there are numerous factors which can determine the effectiveness. The present study identifies some of these factors and provides evidence based recommendations for the improvement of school screening effectiveness. After reviewing all the research papers which originated from the Thriasio school screening program and published in peer review journals, specific suggestions for the organization, the optimal age of screening according to the geographical latitude, the best examined position, the standardization of referrals, the follow up of younger referrals with trunk asymmetry and the reduction of the financial cost are made. We strongly suggest the introduction of these recommendations to all the existing school screening programs in order to improve their effectiveness and to reduce the negative impact they may have on families and on the health system. PMID- 18401086 TI - Clinical evaluation of scoliosis during growth: description and reliability. AB - The clinical evaluation, even today, remains a central point in the diagnosis, prognostic definition and treatment prescription regarding scoliosis. The clinical evaluation of a scoliotic patient has been established for a long time, but it has not been standardized. The aim of the present work is to report the most common clinical measures for the assessment of scoliosis, explain the usefulness of each clinical measurement, and report the repeatability and limits in order to help the physician in making appropriate clinical choices. METHODS: The height of the hump, the angle of trunk rotation, the sagittal and frontal profiles, and the Trunk Aesthetic Clinical Evaluation (TRACE) have been fully described, and their reliability and repeatability have been assessed. RESULTS: The measures analyzed showed good reliability and repeatability on the intra operator basis. The inter-operator repeatability is usually not that good. CONCLUSION: The main measures of the clinical assessment of scoliotic patients have been tested, and their reliability has been evaluated. The knowledge of measurement error, as well as intra- and inter-operator reliability, are essential for the clinical evaluation and treatment of scoliosis. This is an unavoidable basis for decision making in the assessment and the treatment of scoliosis. PMID- 18401087 TI - 3-DEMO classification of scoliosis: a useful understanding of the 3(rd) dimension of the deformity. AB - The third-dimension of scoliosis represent a great challenge for clinicians used to think in two dimensions due to the classical radiographic representation of the deformity. This caused problems in everyday clinical approaches, and led to the development of new bidimensional classifications (King, Lenke) who tried in different ways to face these problems, mainly in a surgical perspective. Recently, some three-dimensional classifications have been proposed, all developed in laboratory by bioengineers. In this paper we present the existing classifications of scoliosis, both bi-dimensional and three-dimensional and we thoroughly discuss the 3-DEMO (3-D Easy Morphological) that has been first presented years ago, and recently thoroughly published; this classification has been developed by clinicians with the main aim of being understandable and easily applicable to everyday clinical life. PMID- 18401088 TI - Inclusion and assessment criteria for conservative scoliosis treatment. AB - The efficacy of brace or conservative treatment in adolescent idiopathic scoliosis is controversial due to variations in inclusion and assessment criteria. This makes the interpretation of brace studies and their comparisons difficult. The Scoliosis Research Society recently introduced new standardized inclusion and assessment criteria for future brace studies. The inclusion criteria include: age 10 years or older at initiation of bracing, Risser sign 0 2, primary curve magnitude 25 to 40 degrees, no prior treatment, and females either premenarche or less than one year post-menarche. The assessment criteria include: percentage of patients with < or = 5 degree curve progression and percentage of patients with > or = 6 degree curve progression at skeletal maturity, percentage of patients who had surgery or recommended before skeletal maturity, percentage of patients with curves exceeding 45 degrees at maturity, and a minimum of 2 years follow-up beyond skeletal maturity for those patients felt to have been successfully treated. All patients treated irregardless of compliance are to be included in the results (intent to treat). The use of these criteria should assist in the determination of the effectiveness of brace treatment, as well as accurate comparison between patient groups and different braces. PMID- 18401089 TI - Indications for conservative management of scoliosis (SOSORT guidelines). AB - This guideline has been discussed by the SOSORT guideline committee prior to the SOSORT consensus meeting in Milan, January 2005 and published in its first version on the SOSORT homepage: http://www.sosort.org/meetings.php. After the meeting it again has been discussed by the members of the SOSORT guideline committee to establish the final 2005 version submitted to Scoliosis, the official Journal of the society, in December 2005. This chapter is a republication from the original paper published in "Scoliosis" BioMed journal and it is included in this book due to its high importance. PMID- 18401090 TI - Specific exercises in the treatment of scoliosis--differential indication. AB - Different methods of physiotherapy are applied in scoliosis management and different opinions exist about the efficacy of conservative scoliosis treatment. Because this divergence of opinions corresponds to a great variety of standards applied, it is not surprising that also the results of conservative treatment greatly differ. Scoliosis normally does not have such dramatic effects that immediate surgery would be indicated. Moreover it is clear that functional and physiological impairments of scoliosis patients-including pain, torso deformity, psychological disturbance and pulmonary dysfunction-require therapeutic intervention. The triad of out-patient physiotherapy, intensive in-patient rehabilitation (SIR) and bracing has proven effective in conservative scoliosis treatment in central Europe. Indication, content and results of physiotherapy are described and discussed in this paper. The differential indication of methods of physiotherapy assigned to current "Best Practice" is documented here as well. The positive outcome of current "Best Practice" conservative management validates a policy of offering conservative treatment as an alternative to scoliosis patients, including those for whom surgery is discussed. PMID- 18401091 TI - Scientific Exercises Approach to Scoliosis (SEAS): efficacy, efficiency and innovation. AB - SEAS is an acronym for "Scientific Exercises Approach to Scoliosis". Main characteristics of SEAS are team approach and cognitive-behavioural approach because in our view these are two indispensable elements in chronic disease rehabilitation. In this article we describe the main differences between SEAS approach and other exercise techniques as well as theoretical bases and therapeutic goals. We illustrate practical application of SEAS concept and scientific results in order to reduce the patient's progress of scoliosis so that a brace would be needed. When compared to usual care, improvement of scoliosis parameters and balance normalization in scoliosis patients. PMID- 18401092 TI - Scoliosis intensive out-patient rehabilitation based on Schroth method. AB - Conservative management of idiopathic scoliosis (IS) and other spinal deformities is a real alternative to surgical treatment. Most of adolescent with IS can be managed conservatively with high safety. Many infantile and juvenile cases show also a good immediate response to conservative care, which can be considered a sign of good prognosis. Only patients showing a continue deterioration even treated conservatively with efficient techniques should be considered candidates to surgical correction and stabilization. Rehabilitation (including specific exercises) and bracing are usually involved in conservative care of IS. In this paper we describe our personal approach in conservative scoliosis care regarding rehabilitation. Bracing has been described in a different paper also published in the present book. Specific exercises can change the signs and symptoms in scoliosis patients. Specialists in physiotherapy for spinal deformities teach the patient how to perform a routine of 'curve pattern' specific exercises with the purpose to facilitate the correction of the asymmetric posture and to teach the patient to maintain the corrected posture in daily activities. Principles of correction are based on those developed by the German physiotherapist K. Schroth. PMID- 18401093 TI - Dobosiewicz method physiotherapy for idiopathic scoliosis. AB - The method developed since 1979, comprises active 3-dimensional auto-correction, concerning the primary curve mobilization towards the correction of the curvature, with special emphasis on the kyphotization of the thoracic spine, carried on in closed kinematic chains, and developed on a symmetrically positioned pelvis and shoulder girdle, followed by active stabilization of the corrected position, and endured as postural habit. The positions for exercising and the movements involved are described in details. Small, moderate and important curves can be managed with DoboMed, however the effectiveness of the therapy depends on the curve flexibility and patient's compliance. DoboMed has been used as a single therapy or together with bracing, as well as preparation for scoliosis surgery. The published results demonstrated that the DoboMed has a positive influence on inhibition of the curve progression in idiopathic scoliosis, the improvement of respiratory functions, assessed by the spirometric values, and the general exercise efficiency evaluated using ergospirometry. PMID- 18401094 TI - Function of the respiratory system in patients with idiopathic scoliosis: reasons for impairment and methods of evaluation. AB - The paper presents the review of pathological changes which develop within the respiratory system in patients with structural progressive idiopathic scoliosis. The impairment of the function of the respiratory system is one of the principal impact of idiopathic scoliosis on the general health and function, as well as on the quality of life. Although the fatal outcomes of respiratory failure are usually prevented by a successful conservative treatment or by the spinal surgery, the reduction of the volume of the thorax, the restriction of the thorax, as well as decreased efficacy of the respiratory muscles are still a major issue (problem) for many patients with structural scoliosis that may lead to respiratory insufficiency or failure. The papers presents main functional tests to assess the respiratory impairment and the basic rules for interpretation of the results of the examination. PMID- 18401095 TI - Side-shift exercise and hitch exercise. AB - We use side-shift exercise and hitch exercise for the treatment of idiopathic scoliosis. Outcomes of side-shift exercise used for the curves after skeletal maturity or used in combination with part-time brace wearing treatment are better than the natural history. Side-shift exercise and hitch exercise are useful treatment option for idiopathic scoliosis. PMID- 18401096 TI - "FITS" concept Functional Individual Therapy of Scoliosis. AB - Functional Individual Therapy of Scoliosis--FITS concept may be used as a separate system of scoliosis correction, a supportive therapy to bracing, children preparation to surgery and also shoulder and pelvic girdle correction after surgical interventions. Taking into account the dysfunctions accompanying scoliosis, the authors of the concept propose an individually adjusted programme of exercises depending on a curvature angle and a result of clinical examination of a patient. On this basis both general and specific goals are set. FITS concept consists of two stages: --Elimination of myofascial restrictions which limit a three-plane corrective movement, by using different techniques of muscle energization; --building new corrective posture patterns in functional positions. By sensory motor balance training and exercising the lower trunk stabilization we can start teaching corrective breathing (with the scoliosis convexities) and adjust corrective movement patterns (in open and close chains). A selection of corrective movement patterns will depend on a type of scoliosis, a direction of spine rotation and a place of building functional compensation. Every corrective pattern includes three components: flexion/extension (the saggital plane correction), elevation/depression (the frontal plane correction) and external/internal rotation (the transverse plane correction). In the processs of therapy the corrective movement patterns are being changed depending on curvature angle behaving and clinical picture of a patient. PMID- 18401097 TI - Brace treatment for adolescent idiopathic scoliosis. AB - Adolescent idiopathic scoliosis (AIS) presents as an abnormal curvature of spine with vertebral rotation. It may impact the patient for their entire life. Approximately 0.25% of adolescents will require treatment. Brace treatment is the most commonly used non-surgical treatment for AIS. Its goal is stop the progression and maintain the curve at an acceptable level through the high risk growth phase of adolescence. However, its effectiveness is controversial and the actual biomechanical action of the brace is not fully understood. Recently, the Scoliosis Research society (SRS) created a standard criterion for AIS brace studies. However, to evaluate the effectiveness of the brace treatment, the spine flexibility, in brace correction and patient's compliance should also be included in a study. Although bracing has been used for more than 50 years, there are still many unknowns. How much wear time per day is needed for an optimum treatment outcome? How much brace tightness is optimal? What is the best weaning protocol? What is the best method to determine the curve flexibility? How much in brace correction is needed to obtain good results? Without accurate and precise methods to objectively measure or answer the above questions, it is misleading to state whether or not brace treatment is effective. Therefore, a lot of research is still required before one can answer the effectiveness of the brace treatment. PMID- 18401098 TI - Biomechanical and clinical perspectives on nighttime bracing for adolescent idiopathic scoliosis. AB - The present review article aims at providing an update on the basic science and clinical information underlying the use of nocturnal braces for adolescent idiopathic scoliosis. The use of nocturnal braces has been dictated by the encouraging results recorded by some studies on part-time bracing, combined with increasing concerns on poor patient compliance noted with the use of full-time bracing. The cardinal feature of nighttime braces lays in their ability to hypercorrect the scoliotic curvature, thereby eliminating the asymmetric water accumulation that occurs in the apical and adjacent intervertebral discs, thus restoring a close-to-normal force application through the Hueter-Volkmann principle and preventing curve progression. The two nighttime braces mostly used hypercorrect the spine through different mechanisms, one acting by bending the spine and the other by the application of opposing forces. Based on the clinical results available, nighttime braces constitute an attractive option for single major lumbar/thoracolumbar curves not exceeding 35 degrees in magnitude. Multi center, randomized studies using strict criteria set forth by the Scoliosis Research Society and the Study group On Scoliosis Orthopaedic and Rehabilitation Treatment are needed to better define the role of nocturnal bracing in the conservative treatment of adolescent idiopathic scoliosis. PMID- 18401099 TI - The cheneau concept of bracing--actual standards. AB - In-brace correction and compliance are the main predictors of a successful outcome of brace treatment in the management of patients with Idiopathic scoliosis. The latest CAD/CAM or module based bracing concepts, related to a proper classification have lead to a better in-brace correction and have made the braces easier to wear for the patient. Nevertheless, the latest developments on the market do not allow successful treatment in every case. The latest biomechanical models of brace correction therefore may lead to a differential indication for certain concepts described in this paper. Thoracic curves with Cobb angles < 50 degrees may be treated with the best possible success with the latest Cheneau derivates enabling a real 3D-correction including also the sagittal correction of the spine. The application of those braces demands a proper classification of curve patterns. Thoracic curves with Cobb angles > 50 degrees demand to increase the force vector from dorsal with the ventral counteraction of subclavicular pads both sides, although this may be at the cost of sagittal correction. The percentage of in-brace correction is a good indicator for brace action, however in the individual case this is not always the most important factor. PMID- 18401100 TI - The Cheneau concept of bracing--biomechanical aspects. AB - Current concept of bracing must take in consideration both the three-dimensional (3D) nature of Adolescent Idiopathic Scoliosis (AIS) and its pathomechanism of progression. A modern brace should be able to correct in 3D in order to break the so called 'vicious cycle' model. Generally speaking, it is necessary to create detorsional forces to derotate in the transversal plane, to correct the lateral deviation in the frontal plane and to normalize the sagittal profile of the spine. Breathing mechanics can be used to fight against the thoracic structural flat back. The original Cheneau brace was introduced at the end of the 70's and its principles were based more in anatomical observations rather than in biomechanics. A further evolution , enunciating new principles, has allowed a higher standard, improving in brace corrections and trunk modelling. This biomechanical principles have been developed under the name of Rigo-Cheneau System (RSC) and used later in latest brace models like the Cheneau light with reduced material, and similar in brace corrections. Experience is also important to improve the end results. The blueprints to built the brace according to the anatomorradiological pattern are very helpful. PMID- 18401101 TI - Passive and active mechanisms of correction of thoracic idiopathic scoliosis with a rigid brace. AB - Contemporary bracing developed numerous novel technical solutions to most of main aspects of the correction of structural progressive idiopathic scoliosis. This paper presents a short review on principal biomechanical rules for the three dimensional scoliosis correction. Apart from the tissue transfer, which is a known passive mechanism of rigid bracing, the other passive mechanisms are described, containing the "cherry stone" distraction effect, the thoracic derotation and the bending. A demanding technical construction of the orthosis enables active mechanisms to develop: the corrective factor of the vertebral growth, hypercorrection-oriented trunk movements and respiration, as well as the anti-gravitational mechanism, by which postural reflexes maintain the curve correction, proximally and distally out of the limits of the brace. We believe that systematic investigations unfolded in the area of neurophysiological aspects of postural control of the spinal balance will continuously improve the fascinating capabilities of the active scoliosis autocorrrection assisted by the brace. PMID- 18401102 TI - Lyon brace. AB - For the last 60 years, the impressive progress of the scoliosis surgery has hidden the development of the conservative orthopedic treatment. The stabilization of the scoliosis, which implies the safeguarding of a spine as mobile as possible, remains a valid objective. The Lyon Brace management combines 3 techniques. A reduction of the scoliosis using a plaster cast fixed on an EDF (Elongation Derotation Flexion) Cotrel's frame. It carries through a flow of the musculoligamentar structure of the concavity. A contention by Lyon Brace. Orthesis without any cervical superstructure is adjustable, symmetric, see through and active. The elongation between the two scapular and pelvic girdle leads to a disc decompression which makes easier the 3D correction of the curves. The individual moulding (custom made) is actually electronic using a "full 3D imaging" system by Orten. To every 14 types of Lenke's classification matches a specific blue print. A specific physiotherapy combining the consciousness of the deformity, suppling up of the retracted elements of the concavity, compensatory suppling up of the girdles, improvement of the vital capacity based on exhalation, reharmonisation of the static, static strengthening in order to facilitate the ability to be still in a corrected position, kyphotisation proprioceptive exercises to stimulate the maturation of the postural system. We advise the scoliotics to practice sport during the treatment period. The long term follow up confirms a global effectiveness indication of 0,89 with the rib hump declining by half. When we treat scoliotics with Cobb angle less than 45 degrees, surgical treatment can be prevented in 98% of the patients.In France 60% of the families agree with this stringent treatment which becomes easier thanks to its ambulatory realization and the excellent formation of the partners, the physiotherapists and the orthesist. PMID- 18401103 TI - Treatment of early adolescent idiopathic scoliosis using the SpineCor System. AB - The purpose of this prospective observational study was to evaluate the effectiveness of the Dynamic SpineCor System for adolescent idiopathic scoliosis in accordance with the standardized outcome criteria proposed by the Scoliosis Research Society Committee on Bracing and Nonoperative Management. The SpineCor System is the first and only truly dynamic brace, which provides a progressive correction of Idiopathic Scoliosis from 15 degrees Cobb angle and above. The new therapeutic approach is based on a new concept upon the etiology and pathogenesis of idiopathic scoliosis; a pathology of the neuro-musculoskeletal system in growth and maturation. This prospective observational study was carried out on a group of 639 patients (92.3% females) having idiopathic scoliosis treated with the SpineCor brace. Five hundred and eighty three patients met the criteria for inclusion, and 234 patients were still actively being treated. Overall, 349 patients have a definitive outcome. All girls were premenarchal or less than 1 year postmenarchal. Assessment of brace effectiveness included (1) percentage of patients who have 5 degrees or less curve progression, and percentage of patients who have 6 degrees or more progression; (2) percentage of patients who have been recommended/undergone surgery before skeletal maturity; (3) percentage of patients with curves exceeding 45 degrees at maturity (end of treatment); and (4) Two-year follow-up beyond maturity to determine the percentage of patients who subsequently underwent surgery. Successful treatment (correction, > 5 degrees, or stabilization, +/- 5 degrees) was achieved in 259 (74.2%) of the 349 patients from the time of the fitting of the SpineCor brace to the point in which it was discontinued (or at the time of the surgery). Fifty one immature patients (14.6%) required surgical fusion while receiving treatment. Eight mature patients out of 298 (2.7%) required surgery within 2 years of follow-up beyond skeletal maturity. The conclusion drawn from these findings is that the SpineCor brace is effective for the treatment of adolescent idiopathic scoliosis. Moreover, positive outcomes are maintained after 2 years because 151 (93.2%) of 162 patients stabilized or corrected their end of bracing Cobb angle up to 2 years after bracing. PMID- 18401104 TI - The SPoRT (Symmetric, Patient-oriented, Rigid, Three-dimensional, active) concept for scoliosis bracing: principles and results. AB - The biomechanical action of an orthesis for the conservative treatment of AIS has two goals: correction and stabilization. These goals have been pursued through very well established principles of correction, developed over the years, divided in terms of efficacy (the correct positioning of pushes, as well as through escape ways and proper drivers of the forces and stops) and acceptability (compliance, perfect body design, maximal freedom in the ADL). To achieve all these goals, the Sforzesco brace has been developed through progressive changes and verification. Finally, we discovered we had something new, and summarised it in the SPoRT acronym: Symmetric, Patient-oriented, Rigid, Three-dimensional, active. The SPoRT concept always requires a customised construction of the brace according to the patient's individual requirements. It's possible to apply CAD CAM technologies, which usually allow us to obtain the best results in this case, but without using pre-built forms stored in databases, as is usually done. Once done, a final test must be made on the patient so as to change the first theoretical project and adapt it in the best possible way, depending on the real interaction between the body and the brace. The results that are today available on the SPoRT concept relate to the Sforzesco brace and necessarily are short term, because the first treated patients are now reaching the third-year follow up examination and haven't yet completed their treatments. According to first studies we can state that: tte Sforzesco brace is more effective than the Lyon brace after six months of treatment; the Sforzesco brace is equally effective as Risser Plast brace. PMID- 18401105 TI - The Boston Brace System philosophy, biomechanics, design & fit. AB - The Boston Brace System developed in the early 1970's by Dr John Hall and Bill Miller, CPO, is a logical, multidisciplinary approach to the treatment of idiopathic scoliosis. The Brace itself is but one component of the Brace system. The clinical team consists of patient, family, orthopedist, orthotist, physical therapist and nurse. Each team member needs to have a working knowledge of each others discipline, and educate the patient on his/her roll of the treatment plan. If the patient is not educated and understands the process, the logic behind the process and the critical roll they play- then the whole system has been compromised. The Boston Brace itself is one of the most widely studied orthosis used in the conservative management of scoliosis. It has been shown that orthotist training and skill, as well as the ability to assess and modify the fit in 3D have a positive influence on patient outcome. This chapter will discuss the philosophy, biomechanics, design, fit and adjustments necessary for a successful outcome. By following the basic tenants of the system, and maintaining the patient focus, the goal of having a stable spine in adulthood can be obtained. It takes a team effort. This outline will take about how to construct and evaluate the orthosis to maximize fit and function. PMID- 18401106 TI - Cosmetic outcome after conservative treatment of idiopathic scoliosis with a dynamic derotation brace. AB - Improved cosmesis is a major concern for the adolescents with Idiopathic Scoliosis (IS). We hypothesized that if we correct the rotation simultaneously to the lateral curvature of the spine with a dynamic brace we may decrease the asymmetry of the back and ultimately improve the cosmetic appearance of the scoliotic child. Thirty six scoliotic children (32 girls and 4 boys) with a mean age of 13.9 (range 12-17) years, a mean Cobb angle of 28.2 degrees (range 19-38 degrees ) and a mean ATI 7.8 degrees (range 4 degrees -17 degrees ) were studied. The examined children were divided in 3 subgroups according to the curve type. All children wore the Dynamic Derotation Brace (DDB), which is a modified Boston brace with antirotatory blades, for 23 hours per day, for a minimum duration of 2 years. The ATI was assessed using the Pruijs scoliometer at baseline and at the end of treatment. ATI was improved statistically significant in the thoracolumbar (p < 0.01) and lumbar region (p < 0.013) of double curves and in the thoracolumbar (p < 0.018) and lumbar region (p < 0.027) of thoracolumbar curves. ATI improvement in the thoracic region was not statistically significant either in double curves (p < 0.088) or in thoracolumbar curves (p < 0.248). For right thoracic curves, ATI improvement was not statistically significant for all the examined regions. The above findings indicate that in double and thoracolumbar curves, a deforming torsional force is present, blocked by the antirotatory action of the blades of the DDB, and seems to be more active in the "lower" spine. In conclusion, DDB improves the cosmetic appearance of the back of IS children with all but right thoracic curves. PMID- 18401107 TI - End-growth results of bracing and exercises for adolescent idiopathic scoliosis. Prospective worst-case analysis. AB - BACKGROUND: In the literature the rate of surgery for AIS (Adolescent Idiopathic Scoliosis) of 30 degrees ranges from 22.4% to 31% when braces are used, versus the natural history rate of 28.1%. When a complete conservative approach is used (braces and exercises), this rate decreases to the range of 3.8% to 7.3%. All these studies are retrospective. AIM: The aim was to evaluate the final results of a prospective set of patients treated in a center fully dedicated to a complete conservative treatment (exercises and braces) of AIS. MATERIALS AND METHODS: This is an everyday clinical, retrospective study on a prospective data base. The population included 112 AIS patients, 13.2+/-1.8 years old, with 23.4+/ 11.5 degrees Cobb degrees at the start of treatment. All the patients had been treated with a full set of conservative treatments, including exercises, according to their individual needs. We used the SEAS (Scientific Exercises Approach to Scoliosis) protocol and the ISICO approach, while the orthosis used included: Risser cast, and the Lyon, Sforzesco-SPoRT, Sibilla-Cheneau and Lapadula braces. The patients had been followed up by the same physician, braces had been made and exercises had been applied by the same team. The outcomes were established for each single patient: The absolute aim was to avoid surgery, while the minimal and optimal outcomes were defined according to the starting curve. An efficacy analysis and worst-case analysis had been performed. RESULTS: The rate of surgery was 0.9% (efficacy analysis), and 4.5% (worst case); the minimal outcomes had been obtained in 99% of patients and the optimal ones in 84%. Overall, the curves over 40 degrees , which numbered eleven at the start of observation, were reduced to three. In total, eight patients exited the presumable area of risk in adulthood (final curve over 30 degrees ). The treatment produced a statistically significant reduction in the worst curves, and the best results have been obtained in the curves over 40 degrees. CONCLUSION: Provided the use of a complete conservative approach, there is very little doubt that it is possible to reduce the rate of surgery in AIS treatment. PMID- 18401108 TI - Quality of life after conservative treatment of adolescent idiopathic scoliosis. AB - As an important consideration in medical care, health-related quality of life (HRQoL) refers to the patient's ability to enjoy normal life activities. When assessing the effectiveness of conservative treatment of AIS, HRQoL variables are more important than the radiographic results or pulmonary function. The present study examines the impact of conservative treatment in HRQoL of the AIS patients. Thirty-two female patients with a mean age of 13.5 (range 12-16) years, a mean Cobb angle of 29.4 degrees (range 21-38 degrees ) and a mean angle of trunk inclination (ATI) 7.9 degrees (range 5 degrees -18 degrees ) were treated with a full time application of a modified Boston brace and occasionally physiotherapy, for a minimum of 2 years. HRQoL was measured with the Brace Questionnaire (BrQ) at baseline and at the end of treatment. Correlations were determined by the Pearson correlation coefficient, with p < 0.05 considered significant. AIS patients scored lower in all the domains of BrQ at the end of treatment. This difference was statistically significant for the mean overall score of BrQ and for the domains of general health perception, physical functioning, emotional functioning, self-esteem and aesthetics, bodily pain and social functioning. The scores in the domains of vitality and school activity were not affected. The HRQoL immediately after the end of conservative treatment of AIS is found to have been negatively affected. Because of limitations in study design, this finding does not necessarily implicate the conservative treatment itself, but highlights the importance of HRQoL measurement in assessing how AIS patients perceive the impact of their disease. PMID- 18401109 TI - A systematic review of analytical observational studies investigating the association between cardiovascular disease and drinking water hardness. AB - The aim of this study is to systematically review and critically assess analytical observational epidemiology studies investigating the association between levels of drinking water hardness and cardiovascular disease. We searched electronic databases and used standardised forms to extract data and assess study quality. Of 2,906 papers identified, 14 met the inclusion criteria (nine case control and five cohort studies). Of the nine case control studies, seven examined both drinking water magnesium and calcium and risk of death from cardiovascular disease. A pooled odds ratio showed a statistically significant inverse association between magnesium and cardiovascular mortality (OR 0.75 (95%CI 0.68, 0.82), p < 0.001). Only two studies reported a statistically significant effect for calcium. Substantial heterogeneity between studies made calculation of a summary estimate for drinking water calcium inappropriate. Of three cohort studies reviewed, two were of good quality. A weak suggestion that soft water was harmful in females and possibly associated with a slightly greater risk of sudden death was reported, but there was no association between water hardness and mortality from stroke or cardiovascular disease. This study found significant evidence of an inverse association between magnesium levels in drinking water and cardiovascular mortality following a meta-analysis of case control studies. Evidence for calcium remains unclear. PMID- 18401110 TI - Large-scale freshwater microbiological study: rationale, results and risks. AB - A fifteen-month fortnightly survey of microbial health risk indicators and pathogens has been carried out at 25 freshwater recreational and water supply sites distributed throughout New Zealand, for: E. coli, Clostridium perfringens spores, F-RNA bacteriophage, somatic coliphage, human enteroviruses, human adenoviruses, Cryptosporidium oocysts, Giardia cysts, Salmonella and Campylobacter. Sites were selected to represent five geographical areas covering New Zealand and five categories of predominant environmental impact: birds, dairy farming, forestry/undeveloped, municipal, and sheep/pastoral farming. Six of the sites were also source waters for treated drinking-water supplies. Of the indicators, E. coli was detected in 99 % of all samples, with somatic coliphage being detected most of the time (89 %). Of the pathogens tested, Campylobacter and human adenoviruses were inferred to be the most likely to cause human waterborne illness to recreational freshwater users. Using data from all sites, an estimated 5 % of notified campylobacteriosis cases in New Zealand could be attributable to water contact recreation. The critical value for E. coli as an indicator of increased Campylobacter infection is in the range of 200-500 E. coli per 100 ml. This result has been used to derive new national water quality guidelines for recreational fresh water in New Zealand. PMID- 18401111 TI - Quantification of the health risk associated with wastewater reuse in Accra, Ghana: a contribution toward local guidelines. AB - Quantitative Microbial Risk Assessment (QMRA) models with 10,000 Monte Carlo simulations were applied to ascertain the risks of rotavirus and Ascaris infections for farmers using different irrigation water qualities and consumers of lettuce irrigated with the different water qualities after allowing post harvest handling. A tolerable risk (TR) of infection of 7.7 x 10(-4) and 1 x 10( 2) per person per year were used for rotavirus and Ascaris respectively. The risk of Ascaris infection was within a magnitude of 10(-2) for farmers accidentally ingesting drain or stream irrigation water; approximately 10(0) for farmers accidentally ingesting farm soil and 10(0) for farmers ingesting any of the irrigation waters and contaminated soil. There was a very low risk (10(-5)) of Ascaris infection for farmers using pipe-water. For consumers, the annual risks of Ascaris and rotavirus infections were 10(0) and 10(-3) for drain and stream irrigated lettuce respectively with slight increases for rotavirus infections along the post-harvest handling chain. Pipe irrigated lettuce recorded a rotavirus infection of 10(-4) with no changes due to post harvest handling. The assessment identified on-farm soil contamination as the most significant health hazard. PMID- 18401112 TI - Impacts of rainfall on the water quality of the Newport River Estuary (Eastern North Carolina, USA). AB - The Newport River Estuary (NPRE), an important North Carolina (NC) shellfish harvesting area, has been experiencing alterations to the land-water interface due to increasing population and coastal development. Water quality degradation in the estuary over the last decade has led to an increase of shellfish harvesting area closures, and has been postulated to be due to non-point source contamination in the form of stormwater. Water samples were taken in the NPRE (n =179) over a range of weather conditions and all seasons from August 2004 to September 2006. Fecal coliform (FC), as estimated by E. coli (EC), and Enterococcus (ENT) concentrations (MPN per 100 ml) were examined in relation to rainfall levels and distance from land. The relationships among the fecal indicator bacteria (FIB) and environmental parameters were also examined. The data revealed a significant increase in FC concentrations after measured rainfall amounts of 2.54 cm (general threshold) and 3.81 cm (management action threshold). However, higher than expected FIB concentrations existed during conditions of negligible rainfall (<0.25 cm), indicating a possible reservoir population in the sediment. Overall, stormwater runoff appears to be adversely impacting water quality in the NPRE. PMID- 18401113 TI - Problems of treatment process selection for relief agency water supplies in an emergency. AB - There is currently limited public information available concerning methods for the selection of appropriate water treatment technologies for application in acute phase emergency water and sanitation responses. Considering the number of available emergency water treatment and supply options, it is speculated that a framework methodology offering guidance in the form of a selection process would offer advantages. This process would relate a defined set of emergency factors to a selection of available water treatment and supply options, described by a basic, but complete, set of technical and operational characteristics. It is suggested that, whilst not a complete process, the proposed framework methodology is sufficient to lead to the identification and selection of appropriate options for deployment. Furthermore the support of critical decision making by a logical framework, could help shape and justify the actions of agencies and workers in the field. PMID- 18401114 TI - Lessons learned from ten investigations of waterborne gastroenteritis outbreaks, France, 1998-2006. AB - Ten outbreaks of waterborne acute gastroenteritis (AGE) have been investigated in France since 1998. These outbreaks have affected populations of over 1,000 people, with generally high attack rates. The causal agents have been identified in six of these events. Aetiologies involved mainly noroviruses and Cryptosporidium sp. The point of entry of the contamination was located in the distribution network in five outbreaks (waste water backflows in four cases and one case of contamination induced by maintenance work) and at the water collection facilities in five other cases. Once the outbreak was detected, epidemiological and environmental investigations and crisis management followed well-established procedures. Further progress in public health surveillance will depend on more complete and rapid detection and reporting. Automated analysis of health insurance data on the reimbursement of drugs for AGE should help make detection more complete. Improved reactivity depends primarily on the operator immediately reporting incidents that indicate a possible massive contamination of the water network to health authorities - in particular complaints from the population, which are the only early-warning alerts in the case of waste water backflows. PMID- 18401115 TI - Bottled water selection and health considerations from multi-element analysis of products sold in New York state. AB - Nineteen bottled water products were purchased from stores in Potsdam and Wappingers Falls, New York and analyzed for 71 inorganic elements by inductively coupled plasma - mass spectrometry (ICP-MS). The wide range in elemental concentrations observed suggests considerable variation in source water composition, processing, and treatment. Comparison with samples from a typical small municipal water system (Potsdam, New York) was made to evaluate the differences between bottled and municipal water and in many cases little difference is apparent. With the exception of one sample of tonic water and one mineral water, all bottled waters tested meet United States Environmental Protection Agency (USEPA) primary standards for drinking water supplies. Ingestion of some of the waters could provide significant percentages of the reference daily intakes (RDI) of key trace elements. Knowledge of the inorganic chemistry of bottled water can help consumers select the brands best suited to their individual health needs or preferences. PMID- 18401116 TI - Inactivation of Cryptosporidium parvum under chlorinated recreational water conditions. AB - Cryptosporidium is a chlorine-resistant protozoan parasite and the etiological agent in many disinfected recreational water outbreaks. While previous studies have reported disinfection Ct values for Cryptosporidium parvum using sodium hypochlorite, these studies have employed conditions and procedures which are not ideal for establishing public health remediation recommendations for chlorinated recreational water venues. In the present study, free chlorine Ct values were measured at pH 7.5 using young oocysts (<1 month old) and tissue culture to determine oocyst viability. Two different oocyst isolates were used: one originating from Iowa and one from Maine (USA). This study determined that the Ct values for a 3-log reduction in oocyst viability were 10,400 (Iowa) and 15,300 (Maine) at pH 7.5. These Ct values are higher than the Centers for Disease Control and Prevention (USA) currently recommends (Ct = 9,600) for achieving a 3.0-log inactivation of Cryptosporidium oocysts during remediation of recreational water venues following fecal diarrhea accidents. PMID- 18401117 TI - An investigation of bacteriological and chemical water quality and the barriers to private well water sampling in a Southwestern Ontario Community. AB - Private well owners in Canada are responsible for maintenance, including routine sampling, of their private drinking water supply. Sampling rates in a Southern Ontario community are well below the public health recommendation. A study with private well owners was conducted to improve private well water sampling rates through the removal of two significant barriers to private well water testing.During the pilot and extended study phases, 549 nitrate and 425 bacteriological water sampling bottles were delivered to private well owners and water samples were collected the following day. A follow-up telephone survey was conducted with both study participants and non-participants to identify barriers to private water sampling that were encountered by the study sample population.Participation rates in the pilot and extended study phases were less than 50% prompting the follow-up telephone survey. Inconvenience and lack of time [statistically significant, P < 0.01] were found to be the main barriers for participation in the study.The findings from this study illustrate the influence that certain barriers have on the frequency of private well water testing in a Southern Ontario community. The findings provide guidance for other health authorities to improve sampling rates. PMID- 18401118 TI - First report of Cryptosporidium spp. oocysts in oysters (Crassostrea rhizophorae) and cockles (Tivela mactroides) in Brazil. AB - The consumption of oysters and cockles, which are usually eaten raw or lightly cooked, can cause outbreaks of human diseases, especially if these shellfish are harvested from polluted areas. In Brazil data about the occurrence of pathogens, like hepatitis A virus, in shellfish have been reported but research on natural contamination for pathogenic protozoa is still non-existent. Cryptosporidium oocyst contamination of oysters (Crassostrea rhizophorae) and cockles (Tivela mactroides) was evaluated during two different periods in a coastal area from Sao Paulo, Brazil. From June to November 2005, and from July to December 2006, 180 mollusks were harvested for tissue examination. The gills and gastrointestinal tract (n = 36 pools) were carefully extracted from the animals and homogenized in a tissue homogenizer by adding surfactant Tween 80 (0.1%). Immunofluorescence assays were performed and Cryptosporidium oocysts were detected in 50.0% of gill pools of cockles and 10.0% of gill pools of oysters. In order to evaluate seawater quality in shellfish growing areas, total levels of thermotolerant coliforms, Escherichia coli and enterococci were determined. This is the first time that Cryptosporidium oocysts were found in shellfish from the coastal region of Brazil, and to the best of our knowledge it is also the first report in Latin America and the case might be of public health importance, reflecting the extension of the contamination on seafood, requiring a need for quality control standards. PMID- 18401119 TI - Increased prevalence of indicator and pathogenic bacteria in Vembanadu Lake: a function of salt water regulator, along south west coast of India. AB - Prevalence of faecal indicator bacteria, Escherichia coli and pathogenic bacteria, Vibrio cholerae, Vibrio parahaemolyticus and Salmonella were analysed in Vembanadu lake (9 degrees 35'N 76 degrees 25'E), along south west coast of India for a period of one year from ten stations on the southern and northern sides of a salt water regulator constructed in Vembanadu Lake in order to prevent incursion of seawater during certain periods of the year. While the northern side of the lake has a connection to the sea, the southern side is enclosed when the salt water regulator is closed. The results revealed the water body is polluted with high faecal coliform bacteria with mean MPN value ranging from 1718-7706/100 ml. E. coli, V. cholerae, V. parahaemolyticus and Salmonella serotypes such as S. paratyphi A, B, C and S. newport were isolated and this is the first report on the isolation of these Salmonella serovars from this lake. E. coli showed highest percentage of incidence (85.6-86.7%) followed by Salmonella (42-57%), V. choleare (40-45%) and V. parahaemolyticus (31.5-32%). The increased prevalence of indicator and pathogenic bacteria in the enclosed southern part of Vembanadu Lake may be resulting from the altered flow patterns due to the salt water regulator. PMID- 18401120 TI - Application of a risk management system to improve drinking water safety. AB - The use of a comprehensive risk management framework is considered a very effective means of managing water quality risks. There are many risk-based systems available to water utilities such as ISO 9001 and Hazard Analysis and Critical Control Point (HACCP). In 2004, the World Health Organization's (WHO) Guidelines for Drinking Water Quality recommended the use of preventive risk management approaches to manage water quality risks. This paper describes the framework adopted by Yarra Valley Water for the development of its Drinking Water Quality Risk Management Plan incorporating HACCP and ISO 9001 systems and demonstrates benefits of Water Safety Plans such as HACCP. PMID- 18401121 TI - Impact of jerry can disinfection in a camp environment - experiences in an IDP camp in Northern Uganda. AB - In July 2007, a study by the Centre for Environmental Health Engineering, at the University of Surrey, assessed a modified method of jerry can cleaning in an internally displaced persons (IDP) camp in Kitgum, N. Uganda. The poor condition of drinking water vessels used in the camp was confirmed as a potential source for microbiological contamination both visually and by microbiological testing. Jerry cans were disinfected using high strength sodium hypochlorite (NaOCl) generated using an experimental AquaChlor Solar unit. The study suggested that regular jerry can cleaning, using a high strength chlorine based disinfectant, offers an effective method of alleviating the adverse effects of contamination in water collection and storage vessels. Results indicated that the method is capable of significantly reducing thermo-tolerant coliform numbers to below 5 cfu/100 ml in most cases. Chlorine strength depletion after repetitive cleaning confirms the impact of process. The method does not substitute for good hygiene practices, which are essential for maintaining water quality in the household. It is suggested that the process can play an important role during outbreaks of water-borne diseases, such as cholera, particularly if combined with regular water disinfection. PMID- 18401123 TI - Water safety plans: planning for adverse events and communicating with consumers. AB - A wide range of microbial and chemical characteristics in drinking water have the potential to affect human health. However, it is not possible or practical to test drinking water for all potentially harmful characteristics. If drinking water is contaminated, people may already be exposed by the time test results are available. The 'boil water alert' issued in Sydney, Australia in 1998 following the detection of Cryptosporidium and Giardia in the finished water supply, highlighted the uncertainties associated with the public health response to test results. The Sydney experience supports the international consensus that a preventive risk-management approach to the supply of drinking water (manifesting as water safety plans (WSPs)) is the most reliable way to protect public health. A key component of a comprehensive WSP is that water suppliers and health authorities must have plans to respond in the case of water contamination and/or outbreaks. These plans must include clear guidance on when to issue warnings to consumers, and how these warnings are to be communicated. The pressure on health authorities to develop clear and systematic boil-water guidance will increase as utilities all over the world develop their WSPs. PMID- 18401122 TI - Resuscitation of eleven-year VBNC Citrobacter. AB - Citrobacter freundii strain WA1 was stressed by incubation in seawater microcosms for eleven years. After two years of starvation, no culturable strain was observed. Incubation of samples in nutrient-rich broth medium not supplemented with growth factors, however, allowed resuscitation of VBNC cells so that subsequent plating yielded observable colonies for significantly extended periods of time. Recovery of VBNC Citrobacter freundii was obtained by incubation in nutrient broth even after eleven years of starvation. To see whether the samples contained the same strain of Citrobacter freundii inoculated 11 years ago. The complete 16S rRNA gene was PCR amplified and sequenced from initial, stressed and revived strains of Citrobacter freundii strain WA1.The 16S rRNA gene sequences from eleven-year stressed strains were homologous with a high degree of similarity to the GenBank reference strain and were identical to each other. PMID- 18401124 TI - Unintentional drinking-water contamination events of unknown origin: surrogate for terrorism preparedness. AB - Drinking-water is a direct conduit to many human receptors. An intentional attack (e.g. terrorism) on drinking-water systems can shock and disrupt elements of national infrastructures. We report on an unintentional drinking-water contamination event that occurred in Tel Aviv, Israel in July, 2001. Initially of unknown origin, this event involved risk management strategies used by the Ministry of Health for abating a potential public health crisis as might be envisaged of water contamination due to terrorism. In an abrupt event of unknown origin, public health officials need to be responsible for the same level of preparedness and risk communication. This is emphasized by comparison of management strategies between the Tel Aviv event and one of dire consequences that occurred in Camelford, England in 1988. From the onset of the Tel Aviv incident, the public health strategy was to employ the precautionary principle by warning residents of the affected region to not drink tap water, even if boiled. This strategy was in contrast to an earlier crisis that occurred in Camelford, England in 1988. An outcome of this event was heightened awareness that a water crisis can occur in peacetime and not only in association with terrorism. No matter how minor the contamination event or short-term the disruption of delivery of safe drinking-water, psychological, medical and public health impact could be significant. PMID- 18401125 TI - Water contamination events in UK drinking-water supply systems. AB - Water supply companies in the UK have a duty under prime UK legislation to notify the Drinking Water Inspectorate of events affecting or potentially affecting the quality of drinking-water supplies. Under the same legislation, the Inspectorate has a duty to investigate each event. After assessing all of the information available, including companies' reports, the Inspectorate advises on the way in which the event was handled and whether any statutory requirements were contravened. If appropriate, a prosecution of the water company may be initiated. Copies of the assessment are sent to the water company, relevant local and health authorities, Ofwat (the economic regulator), the regional Consumer Council for Water and any other interested parties, including consumers who request it. Generic guidance may be issued to the industry on matters of wider concern. This paper considers the role of the Inspectorate, the powers available to it and reporting arrangements. An overview is presented of events that occurred between 1990 and 2005 and common features are identified. Causes of different types of event are discussed. The importance of well-established contacts between the various interested parties involved in protecting public health is emphasised through discussion of example incidents. PMID- 18401126 TI - Making sustainable water and sanitation in the Peruvian Andes: an intervention model. AB - Sustainability of water supplies in remote rural communities is problematic and resource consuming. CARE has a long history of working hand in hand with remote rural communities and devising programs tailored to their needs. We present here an intervention that integrates development of water supplies and sanitation, with operation and maintenance skills development and training of health promoters that can educate from within the community that ensures the sustainability of drinking water supply systems in rural communities. The training used is innovative in that it uses a series of video-workshops which are found to be particularly useful in communities with high illiteracy rates. PMID- 18401127 TI - Drinking-water safety: challenges for community-managed systems. AB - A targeted review of documented waterborne disease outbreaks over the past decades reveals some recurring themes that should be understood by drinking-water suppliers. Evidence indicates the outbreaks are often linked to some significant change in conditions that provides a sudden challenge to a water system. Severe weather events, such as heavy rainfall or runoff from snow melt, as well as treatment process and system changes, are common risk factors for drinking-water outbreaks. Failure to recognise warning signs and complacency are important contributors to drinking water becoming unsafe. Drinking-water suppliers must focus on competence and vigilance in maintaining effective multiple barriers appropriate to the challenges facing the drinking-water system. Understanding the risk factors and failure modes of waterborne disease outbreaks is an essential component for effective management of community drinking-water supplies and ensuring the delivery of safe drinking-water to consumers. PMID- 18401128 TI - Drinking water public right-to-know requirements in the United States. AB - The United States Environmental Protection Agency implements a national drinking water program under the authority of the Federal Safe Drinking Water Act. Amendments to the Act in 1996 added new provisions to enhance consumer understanding of drinking-water issues. Notification requirements associated with annual consumer confidence reports, source water assessments and state compliance reports are intended to enhance the public's knowledge of the quality of their drinking water. Water utilities are also subject to public notification requirements to provide more timely information to consumers in response to violations of health standards. These right-to-know requirements are intended to build the public's confidence, but communicating with consumers can be challenging for both utility managers and government leaders. This paper discusses the need for timely communication, the challenge of providing information when there is uncertainty in the science and the importance of preparing to respond to critical incidents. Because surveys have shown that other members of the community may have better access to consumers or are more trusted, it is important for water utilities to establish relationships with the media and the local public health community. PMID- 18401129 TI - Communicating with healthcare providers. AB - Studies of risk communication have identified healthcare providers, especially physicians, as the source of information most trusted by the public on issues of environmental health. Nothing in medical, nursing or most healthcare provider training actually prepares practitioners to play this role and healthcare providers are generally more oriented toward treatment and medical care than prevention and public health. Healthcare providers require education in order to play this role but rarely seek it. Gaps in the knowledge of professional on the issue of Cryptosporidium illustrate the problem. For members of the professional water community, communicating with healthcare providers is best done when messages are delivered in familiar settings, such as hospital Grand Rounds (a universal format for teaching conferences) and provided in a narrative (case based) form but gaining access is difficult if the topic is not obviously clinical in nature. In addition to being a critically important target group itself, public health professionals are easier to reach and may mediate good working relationships with medical practitioners. We suggest a strategy for water utilities based on partnerships with academic public health and providing education through well-recognized formats in continuing medical and nursing education. PMID- 18401130 TI - Communicating effectively with vulnerable populations during water contamination events. AB - Water contamination events are a public health concern worldwide with significant potential to impact the global community. When communicating with the public during these crisis situations, it is vital to consider the multiple audiences who receive the messages. Before developing or delivering messages to a particular community, it is essential to be familiar with the community's characteristics, needs, concerns, and who is considered credible to that community. Vulnerable populations are those with difficulties in comprehension or accessibility that may limit their full understanding of risks and may mitigate the effectiveness of public health strategies. Vulnerable populations include, but are not limited to, the urban/rural poor, those who are mentally ill, intellectually disabled, medically vulnerable, at the extremes of age (children and the elderly), racial/ethnic minorities, and those with low literacy or limited English proficiency.A water contamination event poses a unique opportunity to work with diverse populations to effectively convey important health messages. Each population needs to receive appropriate public health messages. Becoming familiar with vulnerable populations and their needs prior to a water contamination event will help in identifying barriers and developing and refining effective messages in such a crisis. In water contamination crises, our publics' health depends on effective, targeted crisis communication. PMID- 18401131 TI - The semantics and pragmatics of water notices and the impact on public health. AB - In communicating health risks to the public, it is essential that the message delivered is not misunderstood, as this can lead to risk behaviour. There has been much interest in risk communication and trust, but here we take this further by a cognitive and linguistic analysis of the intended and understood meanings of core words in drinking-water incidents. Based on a questionnaire study of 107 undergraduate UK students using the example of a 'do not drink' notice, we found that, although the majority (87.9%) would buy and drink bottled water, as many as 44% would still drink the polluted tapwater. Males and females would generally behave similarly; however, significantly more men would drink water straight from the tap. All in all, 78% of the population -- males in particular -- could potentially be at risk. We trace the risk behaviour to a misinterpretation of the words drink and not. Public health protection is generally perceived as the choosing of what is safe behaviour and informing the public of appropriate actions in the event of a crisis: however, communicators need to also address the words used to make up risk messages and the prior beliefs that they invoke. PMID- 18401132 TI - Production of thermostable protease enzyme in wastewater sludge using thermophilic bacterial strains isolated from sludge. AB - The volume of sludge produced annually is very high and poses serious disposal problems. The traditional methods of sludge disposal produce secondary pollutants. Therefore, the alternate or suitable solution is reuse of sludge in an ecofriendly approach. Biotechnology is an interesting tool to add value to the processes involved in wastewater and wastewater sludge disposal/reuse. In this context, a study was carried out on thermophilic bacterial strains that produce thermostable proteases. The bacterial strains were first isolated from municipal wastewater sludge. In contrast to the conventional strains used in industries, like Bacillus sp., the new strains were Gram-Negative type. In semi-synthetic medium, a maximal protease activity of 5.25 IU/ml (International Unit per ml) was obtained at a pH of 8.2 and at a temperature of 60 degrees C, which is higher than the stability temperature of 37 degrees C for a similar protease obtained from the conventional producer Bacillus licheniformis. Moreover, growth and protease activity of the strains were tested in wastewater sludge. It is expected that the complexity of sludge could stimulate/enhance the protease production and their characteristics. In conclusion, reuse of wastewater sludge will help to reduce their quantity as well as the value-added products produced will replace chemical products used in industries. PMID- 18401133 TI - Effects of various process parameters on struvite precipitation kinetics and subsequent determination of rate constants. AB - In this paper, struvite (MgNH(4)PO(4).6H(2)O) precipitation kinetics were studied with different operating conditions (varying supersaturation, pH, Mg:P ratio, degree of mixing and seeding conditions) and relevant rate constants were determined by fitting a slightly modified first-order kinetic model to the experimental data obtained. The rate of change of ortho-P concentration in the bulk solutions increases with increasing supersaturation ratio. The estimated rate constants are 2.034, 1.716 and 0.690 hr(-1) for the supersaturation ratio of 9.64, 4.83, and 2.44, respectively. Kinetic parameters were also evaluated for the Mg:P ratio between the ranges of 1.0 and 1.6, indicating higher phosphorus removal efficiency with increasing Mg:P ratio. The rate constants were found to be 0.942, 2.034 and 2.712 hr(-1) for Mg:P ratios of 1.0, 1.3 and 1.6, respectively. The experimental observations for kinetic study of struvite precipitation with different stirrer speeds clearly show that the mixing intensity used had little effect on the intrinsic rate constants. K values found to be 2.034 and 1.902 h(-1) for 100 and 70 rpm, respectively. Seeding, with 250 500 microm of seed crystals during the struvite precipitation kinetics test, was found to have very little effect on the ortho-P removal. PMID- 18401134 TI - Microbial fuel cell cathodes: from bottleneck to prime opportunity? AB - Microbial fuel cells that can generate energy out of wastewaters are close to pilot scale testing. As such, MFC technology is complementary to methane generation due to the possibility to rapidly convert organic acids, polish effluents and work at low substrate concentrations. The main bottleneck perceived at the moment is the cathodic electron transfer. A variety of catalysts has been investigated for the direct transfer of electrons from the cathode to oxygen in the air. Overlooked in this context were bacteria. Bacteria could indeed be worthwhile to replace chemical catalysts. Moreover, their versatility enables us to not only target at oxygen, but also at nitrous oxides and contaminants as possible drivers of electricity generation, nutrient removal and bioremediation. This paper addresses several recent developments in MFC cathode research, and demonstrates that energy generation is but an aspect of this versatile technology. PMID- 18401135 TI - Monitoring and control of the biogas process based on propionate concentration using online VFA measurement. AB - Simple logic control algorithms were tested for automatic control of a lab-scale CSTR manure digester. Using an online VFA monitoring system, propionate concentration in the reactor was used as parameter for control of the biogas process. The propionate concentration was kept below a threshold of 10 mM by manipulating the feed flow. Other online parameters such as pH, biogas production, total VFA, and other individual VFA were also measured to examine process performance. The experimental results showed that a simple logic control can successfully prevent the reactor from overload, but with fluctuations of the propionate level due to the nature of control approach. The fluctuation of propionate concentration could be reduced, by adding a lower feed flow limit into the control algorithm to prevent undershooting of propionate response. It was found that use of the biogas production as a main control parameter, rather than propionate can give a more stable process, since propionate was very persistent and only responded very slowly to the decrease of the feed flow which lead to high fluctuation of biogas production. Propionate, however, was still an excellent parameter to indicate process stress under gradual overload and thus recommended as an alarm in the control algorithm. PMID- 18401136 TI - Process reliability and significance of reclaimed water quality parameters. AB - Microbial quality of reclaimed water can be adequately interpreted by graphical adjustment of experimental results to a lognormal probability distribution; the method serves to verify the degree of conformity to the proposed distribution and to visualize the time variability of the results. A conventional physico-chemical treatment can inactivate up to 1.0 microlog of common bacterial indicators, while maintaining time variability below 1.0 (microlog/100 ml). Dissolved organic matter in the influent will significantly alter the performance and the reliability of disinfection process, highlighting the need for improved adjustments of disinfectant dose to actual quality of treated water. Although those process may reach inactivation rates as high as 6.0 microlog for common bacterial indicators, their reliability may be considerably affected, forcing standard deviations beyond 3.0 (microlog/100 ml). Natural processes can provide inactivation rates from 3.0 to 3.5 for common microbial indicators, under the weather conditions studied. Although those values are lower than those of physical and chemical disinfectants, the reliability of natural processes is more stable, with standard deviations ranging from 0.65 to 1.1 (microlog/100 ml). Natural processes are particularly sensible to external inputs of microbial indicators, due to the presence of wildlife. PMID- 18401137 TI - Investigation of the diversity of homoacetogenic bacteria in mesophilic and thermophilic anaerobic sludges using the formyltetrahydrofolate synthetase gene. AB - Homoacetogenic bacteria are strict anaerobes capable of autotrophic growth on H(2)/CO(2) or CO, and of heterotrophic growth on a wide range of sugars, alcohols, methoxylated aromatic compounds and one carbon compounds, yielding acetate as their sole metabolic end-product. Batch activity tests on anaerobic granular sludge, using H(2)/CO(2) as a substrate and 2-bromoethanesulfonate (BES) as a specific methanogenic inhibitor revealed that H(2)/CO(2) conversion and concomitant acetate production commenced only after a lag period of 60-100 h. This finding suggests that the homoacetogenic population of digester sludge could be maintained by heterotrophic growth on sugars or other organic compounds, rather than by autotrophic growth on H(2)/CO(2). In the present study, two upflow anaerobic sludge bed (UASB) reactors were operated at 37 degrees C and 55 degrees C for two distinct trial periods, each characterised by the application of influents designed to enrich for homoacetogenic bacteria. Specific primers designed for the amplification of the functional gene encoding formyltetrahydrofolate synthetase (FTHFS), a key enzyme in the acetyl-CoA pathway of acetogenesis, were used as a specific probe for acetogenic bacteria. The diversity of acetogens in the granular sludge cultivated in each reactor was revealed by application of FTHFS targeted PCR. Results show that biomass acetogenic composition was dependent upon the operational temperature of the reactor and the substrate supplied as influent. PMID- 18401138 TI - Measurement of H2 consumption and its role in continuous fermentative hydrogen production. AB - To maximise the yield from fermentative H(2) production, H(2) consumption must be minimised. This work demonstrated for the first time that H(2) consumption exists in an established continuous-flow biohydrogen system. The rate of H(2) consumption was found to be related to the concentration of CO(2), with H(2) consumption inhibited at both low and high CO(2). N(2) sparging of the continuous reactor at 31 mL/min/L-liquid increased the H(2) yield from 1.31 to 1.87 mol H(2)/mol glucose, but did not significantly change the in-situ rate of H(2) consumption (0.07-0.09 mM/h). Assuming sparging completely inhibited H(2) consumption, it could only account for 2-11% of the H(2) yield increase during sparging, based on H(2) consumption rates measured in the reactor and in vials. Therefore, H(2) consumption may be of minor concern for continuous biohydrogen systems. PMID- 18401139 TI - Anaerobic digestion of catering wastes: effect of micronutrients and retention time. AB - Source-separated foodwastes collected from a campus catering facility were processed in bench-scale single-stage anaerobic digesters. The feedstock contained a varied mix of fruits, vegetables, meats and fried foods. A constant organic loading rate (OLR) was maintained with differing hydraulic retention times (HRT). Regular addition of trace elements or prolonged retention time allowed stable digestion at high total volatile fatty acid (TVFA) levels. Reactors on HRT of 25, 50, and 100 days with no micronutrient supplementation exhibited methanogenic failure after approximately 40, 100 and 90 days respectively, while duplicate reactors with micronutrient supplementation maintained stable digestion. An extended HRT of 180 days has so far allowed continued digestion (for reactors with and without micronutrient supplementation) at levels of ammonia nitrogen exceeding 5.7 g l(-1) and volatile fatty acid levels exceeding 15 g l(-1), usually considered inhibitory or toxic. PMID- 18401140 TI - Inhibitory effects of nitrate reduction on methanogenesis in the presence of different electron donors. AB - The preferential utilization of different electron donors and their effects on the nitrate reduction and methanogenesis in a mixed, mesophilic (35 degrees C) methanogenic culture were investigated. Batch methanogenic cultures were fed with dextrin/peptone (D/P), propionate, acetate, and H(2)/CO(2) at an initial COD of 500 mg/L and an initial nitrate concentration of 50 mg N/L. Immediate cessation of methane production was observed in all nitrate-amended cultures. Methane production completely recovered in the D/P- and acetate-fed cultures, and partially recovered or did not recover in the propionate- and H(2)/CO(2)-fed, nitrate-amended cultures, respectively. Accumulation of denitrification intermediates was observed in both the propionate- and H(2)/CO(2)-fed cultures, which resulted in inhibition of fermentation and/or methanogenesis. The fastest and the slowest nitrate reduction were observed in the acetate- and propionate fed cultures, respectively. PMID- 18401141 TI - Flow structures in MBR-tanks. AB - In this paper a CFD model is presented that allows for an approximation of the flow field within MBR-tanks. The technique is demonstrated by means of two examples considering both hollow-fibre and sheet-membrane modules. The flow field is computed and a comparison between measured and computed flow velocities shows the good quality of the numerical model. PMID- 18401142 TI - Potentials of using nanofiltration to recover phosphorus from sewage sludge. AB - Due to the depletion of mineral phosphorus resources there is an increasing demand for efficient phosphorus recovery technologies. In this study the potential of nanofiltration to recover phosphorus from pre-treated sewage sludge is investigated. The efficiency of three commercial nanofiltration membranes (Desal 5DK, NP030; MPF34) was tested using model solutions. Desal 5DK showed the best selectivity for phosphorus. A pH of lower than 1.5 was found to be most suitable. Desal 5DK was used on four different sewage sludge ash eluates and on one sewage sludge. In these experiments it was shown that a separation of phosphorus from undesired components such as heavy metals was possible with significant variations in the efficiency for the different ash and sludge types. Additionally the achievable product recovery was investigated with model solutions. A product recovery of 57.1% was attained for pH 1 and 41.4% for pH 1.5. PMID- 18401143 TI - Helminth eggs removal by microscreening for water reclamation and reuse. AB - Irrigation with reclaimed water is becoming a practical alternative to conventional irrigation in semi-arid areas of the Mediterranean like Spain, but it requires a reliable treatment process to provide a safe water supply. Helminth eggs are one of the main concerns for the safe use of reclaimed water, as they can survive adverse environmental conditions and they are highly infective. Spanish water quality criteria and International guidelines set a limit of 0.1 eggs/l for water uses with unrestricted human exposure. Two microscreening processes have been tested to determine their potential for helminth eggs removal, after a conventional physic-chemical reclamation process. Hydrotech Drum and Discfilters, provided with 10 microm pore size filter cloth, were tested to determine their efficiency for helminth eggs straining. An experimental test was conducted using 20 microm spherical latex particles, as surrogates for helminth eggs, to test the removal efficiency of a small full-scale drumfilter. In a subsequent laboratory test, actual Trichuris suis eggs were strained using a 10 microm pore size filter cloth from a discfilter. Results from both tests indicate that drum and discfilters are able to achieve 99% removal efficiency for spherical latex particles and a complete removal for helminth eggs in reclaimed water. PMID- 18401144 TI - Characterization of sludges for predicting anaerobic digester performance. AB - Batch anaerobic digestion tests of primary sludge and waste activated sludge were conducted for a duration of 123 days to determine the ultimate degradability of the sludges. For primary sludges the inert fraction of the particulate COD that was predicted by the wastewater models could be employed to predict their biodegradability under anaerobic conditions. The degradation of waste activated sludge was adequately characterized for the first 60 days of digestion using a model that assumed equivalent biodegradability of particulate COD components under aerobic and anaerobic conditions. However after 60 days of anaerobic digestion it appeared that decay of the endogenous products was occurring. This could be described with a first order decay function with a coefficient of 0.0075 d(-1). For continuous flow digesters operating at SRTs of 30-60 days, the predicted VSS destruction with the modified model was approximately 10% higher than that predicted on the basis of inert endogenous decay products. PMID- 18401145 TI - Effects of treated wastewater irrigation on the dissolved and soil organic carbon in Israeli soils. AB - In many arid and semi-arid regions, the demand for drinking water and other domestic uses is constantly growing due to demographic growth and increasing standard of living. Therefore, less freshwater is available for agricultural irrigation and new water sources are needed. Treated wastewater (TWW) already serves as an important water source in Israel since more than 40 years and its usage will further be extended. Related to its high loads with nutrients, salts and organic materials its use as irrigation water can have major effects on the soil physical, chemical and biological properties, in the worst case leading to soil degradation. Additional organic matter reaches the soil with the effluent water and soil microbial activity is stimulated. Soil organic carbon (SOC) seems to accumulate in the topsoil and tends to decrease after long-term irrigation with secondary TWW in the subsoil. The amount of dissolved organic carbon increased and the aromaticity of the organic compounds in the soil percolates decreased over the irrigation period. Priming effects, occurring after stimulation of microbial activity by the addition of easily degradable substances, could be found in the soils and were stronger for subsoil (1 m depth). PMID- 18401146 TI - Ammonia, a selective agent for methane production by syntrophic acetate oxidation at mesophilic temperature. AB - In biogas processes, methane production from acetate proceeds by either aceticlastic methanogenesis or through syntrophic acetate oxidation (SAO). In the present study, the pathway for methane production from acetate was analysed; i) during a gradual increase of the ammonia concentration (final concentration 7 g NH(4)(+) -N/L) in a semi-continuous lab-scale anaerobic digester (4.3 L), operating at mesophilic temperature (37 degrees C) or ii) in diluted enrichment cultures (100 ml) experiencing a gradual increase in ammonia, sodium, potassium and propionic acid. The pathway for methane formation was determined by calculating the (14)CO(2)/(14)CH(4) ratio after incubating samples with (14)C-2 acetate. In the anaerobic digester, as well as in the enrichment cultures, the (14)CO(2)/(14)CH4 ratio clearly increased with increasing ammonium-nitrogen concentration, i.e. as the ammonia concentration increased, a shift from the aceticlastic mechanism to the syntrophic pathway occurred. The shift was very distinct and occurred as the NH(4)(+) -N concentration rose above 3 g/l. No shift in pathway was seen during increasing concentrations of sodium, potassium or propionic acid. The shift to SAO in the biogas digester resulted in a twofold decrease in the specific gas and methane yield. PMID- 18401147 TI - Cross sectoral and scale-up impacts of greywater recycling technologies on catchment hydrological flows. AB - With the growth of urban areas and climate change, decisions need to be taken to improve water management. This paper reports an assessment of the impact of greywater recycling systems on catchment scale hydrological flows. A simulation model developed in InfoWorks CS (Wallingford Software Ltd) was used to evaluate how river flows, sewer flows, surface runoff and flooding events may be influenced when grey water recycling systems of different number and scale are implemented in a representative catchment. The simulations show the effectiveness of greywater recycling systems in reducing total wastewater volume and flood volume. However, no hydraulic impacts due to implementation of greywater was identified by the model. PMID- 18401148 TI - Integration of equalisation tanks within control strategies for anaerobic reactors. Validation based on ADM1 simulations. AB - The combination of equalisation tanks and anaerobic digesters represents a typical design scenario within the treatment of industrial wastewaters. In this context, if the hydraulic capacity of the equalisation tanks is effectively handled, significant improvements in the performance of anaerobic digesters can be achieved in terms of process stability and biogas production. This paper presents a rule-based control strategy for anaerobic reactors with the objective of maximising in the long-term the net production of biogas. The control algorithm combines real-time information about the state of the anaerobic digester with on-line measurements about the wastewater volume of the equalisation tank in order to set permanently the appropriate production of biogas. Such a strategy guarantees a continuous influent flow so that emptying and overflowing episodes in the equalisation tank can be prevented. Aiming at a further full-scale implementation, only reliable and cost-effective on-line instrumentation has been considered within the control architecture. The performance of the proposed control approach has been validated for an anaerobic hybrid configuration (AHR) by simulation using the IWA ADM1 model. PMID- 18401149 TI - A critical evaluation of combined engineered and aquifer treatment systems in water recycling. AB - Australian experience at five research sites where stormwater and reclaimed water have been stored in aquifers prior to reuse, have yielded valuable information about water treatment processes in anaerobic and aerobic aquifers. One of these sites is the stormwater to potable water ASTR project at the City of Salisbury, a demonstration project within the broader EC project 'RECLAIM WATER'. A framework for characterising the effectiveness of such treatment for selected organic chemicals, natural organic matter, and pathogens is being developed for inclusion in new Australian Guidelines for Management of Aquifer Recharge. The combination of pre-treatments (including passive systems such as reed beds) and aquifer treatment effectiveness in relation to source waters and intended uses of recovered water will be described. Advantages and disadvantages of various types of pre-treatments in relation to effectiveness and sustainability of managed aquifer recharge will be discussed taking account of aquifer characteristics. These observations will be consolidated into a draft set of principles to assist in selection of engineered treatments compatible with passive treatment in aquifers. PMID- 18401150 TI - Heat island mitigation using water retentive pavement sprinkled with reclaimed wastewater. AB - In Japan, reclaimed wastewater has been recycled widely for non-potable urban applications and it is to be used for sprinkling roads to mitigate heat island in urban areas. To assess the heat island mitigation effects of the sprinkling reclaimed wastewater on water retentive pavement, we carried out a survey at Shiodome-District, Tokyo. The temperatures of air and roads, humidity, and WBGT (Wet-bulb globe temperature) were measured and heat flux was estimated to compare the condition of the areas with/without sprinkling. The following results were obtained. 1) Sprinkling reclaimed wastewater decreased the road surface temperature by 8 degrees during the daytime and by 3 degrees at night: temperatures equal to those on planting zones. Nevertheless sprinkling was done only in the daytime, the temperature decrease effect was not only obtained during the daytime: it continued through the night, due to the water retentive pavement. 2) Sprinkling reclaimed wastewater reduced the amount of sensible heat flux and increased that of latent heat flux. These results suggest that sprinkling reclaimed wastewater on water retentive pavement can effectively mitigate the heat island phenomenon. PMID- 18401151 TI - Exerting ultrasound to control the membrane fouling in filtration of anaerobic activated sludge--mechanism and membrane damage. AB - In this study, ultrasound was applied to control membrane fouling development online in an anaerobic membrane bioreactor (AMBR). Experimental results showed that membrane fouling could be controlled effectively by ultrasound although membrane damage may occur under some operational conditions. Based upon the observation on the damaged membrane surface via SEM, two mechanisms causing membrane damage by exerting ultrasound are inferred as micro particle collide on the membrane surface and chemical interaction between membrane materials and hydroxyl radicals produced by acoustic cavitations. Not only membrane damage but also membrane fouling control and membrane fouling cleaning were resulted from these mechanisms. Properly selecting ultrasonic intensity and working time, and keeping a certain thickness of cake layer on membrane surface could be effective ways to protect membrane against damage. PMID- 18401152 TI - Water reuse for urban landscape irrigation: aspersion and health related regulations. AB - The Mediterranean seaside resort of Le Grau du Roi includes 40 hectares of landscaped areas spray irrigated with river water supplied through a separate network. Wastewater collected from several municipalities is treated in an activated sludge wastewater treatment plant (WWTP) and polished in waste stabilization ponds (WSPs). Planned substitution of treated wastewater for river water is hindered by spray irrigation prohibition within a 100 m distance from houses and recreational areas. WWTP and WSP effluents were monitored for pathogens with a particular attention to Legionella in Spring and Summer 2006. Helminth eggs, salmonellae and enteroviruses were never detected neither in WWTP effluent nor in the ponds. Legionella spp content was slightly higher or of the order of magnitude of river water contents. Regarding Legionella pneumophila contents, WSP effluent did not significantly differ from the river water. E.coli and enterococci contents in WSP effluents complied with the "excellent quality" criteria of the European Directive for coastal bathing waters. Therefore, substituting WSP effluents to river water is unlikely to alter health risks related to spray irrigation and, in this case, the buffer zone required by the French water reuse guidelines appears being short of support. PMID- 18401153 TI - Spatial and temporal changes in sulphate-reducing groundwater bacterial community structure in response to Managed Aquifer Recharge. AB - The population dynamics of bacterial able to be cultured under sulphate reducing condition was studied in conjunction with changes in aquifer geochemistry using multivariate statistics for two contrasting Managed Aquifer Recharge (MAR) techniques at two different geographical locations (Perth, Western Australia and Adelaide, South Australia). Principal component analysis (PCA) was used to investigate spatial and temporal changes in the overall chemical signature of the aquifers using an array of chemical analytes which demonstrated a migrating geochemical plume. Denaturing Gradient Gel Electrophoresis (DGGE) using DNA from sulphate-reducing bacteria cultures was used to detect spatial and temporal changes in population dynamics. Bacterial and geochemical evidence suggested that groundwater at greatest distance from the nutrient source was least affected by treated effluent recharge. The results suggested that bacterial populations that were able to be cultured in sulphate reducing media responded to the migrating chemical gradient and to the changes in aquifer geochemistry. Most noticeably, sulphate-reducing bacterial populations associated with the infiltration galleries were stable in community structure over time. Additionally, the biodiversity of these culturable bacteria was restored when aquifer geochemistry returned to ambient conditions during the recovery phase at the Adelaide Aquifer Storage and Recovery site. PMID- 18401154 TI - Sludge deterioration in a full scale UASB reactor after a pH drop working under low loading conditions. AB - A full scale UASB reactor treating the effluent of a malting plant was operated during nearly two years. During 37 weeks of operation the reactor worked with a COD removal efficiency of 80% and a biogas production of nearly 300 m(3)/d with a methane content of 77%. After the start up and during these months of operation the volumetric organic load was 4 kgCOD/m(3).d and the specific organic load was between 0.2-0.4 kgCOD/kgVSS.d. The sludge SMA in this period was around 0.25 kgCOD/kg VSS.d. On week 37 as a result of a problem at the industrial process the pH in the reactor dropped to a value of 4.8. After pH recovering, the reactor worked with fluctuating COD values in the exit and showed a downward trend in the COD removal efficiency. On week 81 the presence of filaments in the granules was detected. High proportion of Chloroflexi filaments were detected by FISH in the sludge. Changes in the microbial population caused by the low pH probably destabilize the reactor performance. The presence of filamentous granules in the sludge and its further growing could be encouraged by the pH drop and the low specific organic load applied to the reactor. The low specific organic load was a consequence of the high VSS content in the UASB reactor, due to the lack of purges. The length of the filaments attached to the granules grew throughout time. In order to eliminate the sludge with poor settlement properties a recycle was applied to the reactor. As a consequence, low amount of granular sludge stayed in the reactor. At the end, COD concentration in the influent reached higher values than in normal operation; at the same time a complete sludge wash out occurred. On the other hand, using the same sludge (after the recycle implementation) in a bench scale reactor the good properties of the sludge were completely recovered. PMID- 18401155 TI - Interleukin-11 for treatment of hepatitis C-associated ITP. AB - BACKGROUND: Immune thrombocytopenic purpura (ITP) is frequently associated with chronic hepatitis C (HpC-ITP). METHODS: Recombinant interleukin-11 (rIL-11), which has both thrombopoietic and anti-inflammatory properties, was evaluated in 12 patients with HpC-ITP in this pilot study. Group 1 (7 patients) was treated at high dose (50 microg/kg daily) while group 2 (5 patients) at low dose (15-35 microg/kg three/week). RESULTS: In group 1, mean platelet counts rose from initial 54 x 10(9)/l to 103 x 10(9)/l (p = 0.02) and in group 2, from an initial 51 x 10(9)/l to 74 x 10(9)/l (p = 0.04). Antiplatelet antibody (aPlt-Ab) decreased in group 1. LFT improved in both groups. The mean HCV-RNA decreased in group 1 (p = 0.04), not in group 2. Side effects were common and troublesome, but were minimized with individualized dosing. One patient achieved good remission of both ITP and HpC lasting >2 years with low-dose maintenance. CONCLUSION: When used based on individual tolerance, rIL-11 appears useful in HpC-ITP. PMID- 18401156 TI - Correlations of sialic acid with markers of inflammation, atherosclerosis and cardiovascular events in hemodialysis patients. AB - BACKGROUND: Serum total sialic acid (S-TSA) is associated with atherosclerotic process in general population. The aim of our study was to evaluate possible correlations of S-TSA with markers of inflammation and atherosclerosis in hemodialysis (HD) patients. METHODS: We involved 53 asymptomatic, nondiabetic HD patients and 28 healthy controls. Atherosclerosis was evaluated by carotid ultrasonography, estimating intima media wall thickness and wall to lumen ratio bilaterally. To confirm our findings, we performed a 32-month cohort study, during which cardiovascular (CV) events were analyzed in relation to S-TSA concentration. RESULTS: HD patients had higher S-TSA compared to controls (adjusted OR: 1.04, p = 0.026). In HD patients, S-TSA independently correlated with hs-CRP (p < 0.0001), lipoprotein(a) (p = 0.02), intima media wall thickness (p = 0.023) and wall to lumen ratio (p = 0.028). Increased S-TSA concentration was associated with more CV events (p = 0.03). CONCLUSIONS: Serum TSA seems to correlate with inflammation, accelerated atherosclerosis and CV events in nondiabetic HD patients, but more studies need to confirm our findings. PMID- 18401157 TI - Proteomics for the investigation of acute kidney injury. AB - Acute kidney injury (AKI), previously referred to as acute renal failure (ARF), represents an important problem in clinical medicine. Despite significant improvements in therapeutics, the mortality and morbidity associated with AKI remain high. The reasons for this include (a) an incomplete understanding of the underlying pathophysiologic mechanisms, and (b) the lack of early markers for AKI, and hence an unacceptable delay in initiating therapy. Fortunately, the application of innovative technologies such as functional genomics and proteomics to human and animal models of AKI has uncovered several novel genes and proteins that are emerging as biomarkers and novel therapeutic targets. Recent advances in proteomics that hold promise in ischemic AKI, the most common and serious subtype of ARF, are chronicled in this article. These include the identification of biomarkers in the plasma (NGAL and cystatin C) and urine (NGAL, KIM-1, IL-18, cystatin C, alpha 1-microglobulin, fetuin-A, Gro-alpha, and meprin) for the investigation of AKI. It is likely that the AKI panels will be useful for timing the initial insult and assessing the duration of AKI. Based on the differential expression of the biomarkers, it is also likely that the AKI panels will distinguish between the various etiologies of AKI, and predict clinical outcomes. PMID- 18401158 TI - Proteomics of plasma and urine in primary nephrotic syndrome in children. AB - Primary nephrotic syndrome in children, especially the variant with segmental glomerulosclerosis, remains an unsolved clinical problem. In spite of some progress, its pathogenesis is still unknown and the therapy options are confined to gross immune modulation. Indirect evidence based on posttransplant recurrence of the disease suggested an implication of plasma factors, whose characterization remains in course. Besides historical candidates, research is now considering glyco- and lipoderivatives. Structural analysis of plasma and urinary proteins based on proteomics has recently shown an increased proteolysis of major components such as albumin and the implication of alpha 1-antitrypsin that represents the first-line defense against exogenous and endogenous substances with proteolytic activity. Albumin has also emerged as a major plasma antioxidant, and recent studies have demonstrated that in patients with active focal segmental glomerulosclerosis albumin undergoes massive and stable oxidation with sulfonation of Cys34, formation of an adduct with +48 Da molecular weight, changes of the net charge due to additional negative residues, and loss of free thiol group (SH) titration. Altogether, these data suggest that oxidative stress determines selective protein damages in focal segmental glomerulosclerosis patients with formation of new adducts and fragmentation of plasma proteins. Research should now address whether oxidation of podocyte proteins is important for the maintenance of renal selectivity and is involved in proteinuria. PMID- 18401159 TI - Urinary proteome profiling to search for biomarkers in steroid-resistant nephrotic syndrome. AB - Long-term outcomes for patients with nephrotic syndrome (NS) correlate closely with the degree of steroid responsiveness. There are currently no diagnostic tests that accurately predict steroid responsiveness in NS. In children in particular, a prolonged course of daily, high-dose corticosteroid therapy is as much a diagnostic maneuver as it is a treatment. Urine proteomics has been emerging as a potentially rich source of noninvasive yet informative biomarkers of drug responsiveness in NS. In this review, we discuss some of the initial studies of the nephrotic urinary proteome as well as some ongoing and future challenges, including defining the normal urinary proteome, and extracting valuable urinary protein data from an abundance of urinary albumin. PMID- 18401160 TI - Searching for novel biomarkers and new therapeutic targets of diabetic nephropathy using proteomics approaches. AB - Diabetic nephropathy is a major complication of diabetes and remains a common health problem worldwide. It is the dominant cause of incident end-stage renal disease. Currently, microalbuminuria is an only noninvasive marker available for the diagnosis of diabetic nephropathy. However, some patients with microalbuminuria have advanced renal pathological changes for which therapy is less effective than earlier stages of the disease. Additionally, the immunoassay to measure microalbuminuria can detect only immunoreactive forms of albumin, whereas immunounreactive forms are undetectable by this conventional method. Therefore, novel biomarkers for earlier diagnosis of diabetic nephropathy are crucially required. Moreover, the incidence rate of diabetes-induced end-stage renal disease remains high despite good control of blood sugar levels and adequate treatment with appropriate regimens, indicating the need of new therapeutic targets for better therapeutic outcome and successful prevention of diabetic nephropathy in diabetes patients. During the proteomic era, proteomics has become a powerful tool for unraveling the disease pathophysiology and for biomarker discovery. This chapter summarizes recent studies that applied proteomics for the investigation of diabetic nephropathy with major aims to search for novel biomarkers and new therapeutic targets. PMID- 18401161 TI - Diagnostic and prognostic biomarkers in acute renal failure. AB - Acute kidney injury (AKI) is a process that can lead to renal failure. No biological markers are available for predicting the cause or prognosis of AKI. Tests that can predict which patients will need renal replacement therapy (RRT) are needed. In this chapter, we review the recent literature for proteomic analysis in AKI and identify new candidate markers to predict the need for RRT. We also used artificial neural network (ANN) analysis of urine protein data obtained by two-dimensional gel electrophoresis from 19 patients with acute tubular necrosis to identify a set of proteins that can predict whether a patient will require RRT. Ten patients were randomly selected to train an ANN algorithm. The remaining 9 patients were withheld to serve as an independent validation set. The ANN algorithm correctly predicted the renal prognosis of all 10 patients in the training set. In the validation set, the test correctly predicted the future course of renal failure in 7 of the 9 patients (78% accuracy) including 3 of 4 patients who would require RRT (75% sensitivity) and 4 of 5 who would not (80% specificity). Combinations of urine proteins can be used to predict which patients will require RRT. PMID- 18401162 TI - Proteomics and renal transplantation: searching for novel biomarkers and therapeutic targets. AB - Renal transplantation has emerged as the preferred option for many patients with endstage renal failure. While significant progress has been achieved in short term outcomes, long-term survival has only marginally improved. Adaptation of immunosuppressive drugs to the individual needs of every patient at every time point after transplant will be essential to improve long-term outcomes. Thus, assays are required that detect allograft injury very early, which implies frequent noninvasive measurements (e.g. in urine or serum). In this review, we describe important general aspects in urine biomarker discovery using proteomics and discuss currently published studies. Although proteomics has the potential to provide insights into complex pathophysiological processes and reveal novel diagnostic biomarkers as well as therapeutic drug targets, the actual status of urine proteomic activities in renal transplantation is still far from reaching these ambitious goals. PMID- 18401163 TI - Metabolomics: a complementary tool in renal transplantation. AB - Renal transplant success is closely tied to the ability to monitor transplant recipients and responsively change their medications. However, transplant monitoring still depends on relatively dated technologies - serum creatinine levels, urine output, and histopathology of biopsy samples. These techniques do not offer sufficient specificity, sensitivity, or accuracy for appropriate and timely interventions. As a result, more specific diagnostic techniques, based on proteomics, genomics and metabolomics are being sought. Metabolomics (the high throughput measurement and analysis of metabolites) may make it possible to monitor transplants more effectively and specifically. Changes in the concentration profiles of a number of small molecule metabolites found in either blood or urine can be used to localize kidney damage, assess organs at risk of rejection, assess kidneys suffering from ischemiareperfusion injury or identify organs that have been damaged by immunosuppressive drugs. The application of metabolomics to kidney transplant monitoring is still in its early stages. Nevertheless, there are a number of easily measured metabolites in both urine and serum that can provide reliable indications of kidney function, kidney injury, and immunosuppressive drug toxicity. Metabolomics could serve as a good complement to existing proteomic and genomic technologies. PMID- 18401164 TI - Understanding and managing renal cell carcinoma: can proteomic studies contribute to clinical practice? AB - Renal cell carcinoma (RCC) is associated with a poor prognosis and there is a need for biomarkers to assist at all stages of disease management including diagnosis, prognosis, monitoring for relapse and predicting response to therapy. Additionally, identification of new therapeutic targets is a priority. Increased understanding of disease pathogenesis and the molecular changes underlying tumour formation is essential to assist in the rational design of such molecules. As the technologies underlying proteomics-based research have developed, they have been applied extensively to the analysis of cancers including RCC, with tissues, cell lines and biological fluids being used for analysis. A number of approaches have been adopted including two-dimensional polyacrylamide gel electrophoresis and mass spectrometry profiling of intact proteins, shotgun mass spectrometry-based profiling at the peptide level, antibody arrays and strategies analysing the immune response to tumours with a view to identifying tumour-associated antigens. Although these studies are still at a relatively early stage, promising results have been reported with some being taken forward to preliminary validation. The challenge now is to build on these initial efforts, focusing particularly on interrogating the less readily accessible, lower abundance proteome and implementing large-scale validation studies to develop potential markers, antigens and targets and facilitate translation of suitable findings into the clinic. PMID- 18401165 TI - Capillary electrophoresis coupled to mass spectrometry for biomarker discovery and diagnosis of kidney diseases. AB - The main focus of urinary proteome analysis in nephrology is currently on detection and identification of polypeptides that significantly alter (in abundance, distribution, etc.) during (patho)physiological changes of the kidney structure and/or function. Capillary electrophoresis coupled online to electrospray ionization time-of-flight mass spectrometry (CE-MS) was applied to human urine to identify biomarkers for clinical diagnostics. To extract the information of the CE-MS spectra in a timely fashion, software was designed to automatically deconvolute and normalize the spectra. Furthermore, bioinformatics and statistic approaches were used to discriminate patients with different diseases and healthy individuals, respectively. Samples from patients with renal diseases display polypeptide patterns that differ significantly from those obtained from healthy individuals. Examining series of patients with the same disease allowed the establishment of polypeptide patterns typical for specific diseases. This permits the search for the disease-specific peptide markers. The combinations of several polypeptides found in urine are forming a specific pattern, which is indicative not only for the particular disease, but also for the stage of disease. These results show that proteome analysis with CE-MS of urinary polypeptides in patients with different renal diseases and urological disorders can display the current status of the kidney or the urogenital tract. This advancement offers the early diagnosis of different nephrological diseases. PMID- 18401166 TI - Identification of urinary biomarkers by proteomics in newborns: use in obstructive nephropathy. AB - An important issue in congenital unilateral ureteropelvic junction (UPJ) obstruction, a frequent pathology in newborns, is whether infants should undergo surgery. Non-invasive biomarkers to reduce or replace the current invasive clinical exploration are not available. The objective of this study was to identify urinary markers of UPJ obstruction. We compared a number of proteome technologies to study the urinary proteome in UPJ obstruction and selected online capillary electrophoresis coupled to mass-spectrometry for the selection of non invasive prognostic biomarkers. We selected 53 urinary biomarkers that were able to distinguish between different levels of UPJ obstruction. In a prospective study using these 53 biomarkers, we predicted with 97% accuracy, and several months in advance, the clinical outcome of 36 UPJ-obstruction patients. Some of the discriminating biomarkers were identified. A newly identified marker, proSAAS (proprotein convertase subtilisin/kexin type 1 inhibitor), generated a new hypothesis in the physiopathology of UPJ obstruction. These results show that analysis of urinary polypeptides in newborns with UPJ obstruction can predict their clinical outcome. PMID- 18401167 TI - Proteomics and kidney stone disease. AB - Kidney stone disease (nephrolithiasis) is an ancient and common affliction. It has been recognized for a long time with evidence of stone found in approximately 7,000-year-old mummies and remains a common problem worldwide, indicating ineffective prevention in the past. Precise pathogenic and molecular mechanisms of kidney stone formation are still poorly understood and should be further elucidated. Also, identification of novel therapeutic targets for better therapeutic outcome and successful prevention of the occurrence and recurrence of the stone are crucially required. One of the most promising tools for current and future biomedical research is proteomics, which has been extensively and widely applied to the nephrology field during the past 5 years. Its high-throughput capability holds a great promise also to kidney stone research. This chapter provides a brief overview of proteomic methodologies recently used for the investigation of nephrolithiasis and recent proteomic studies of nephrolithiasis are summarized. PMID- 18401168 TI - Exploring the uremic toxins using proteomic technologies. AB - Kidney failure leads to the uremic syndrome that is the clinical expression of the malfunction of vital organs due to the accumulation of uremic toxins, which are normally cleared by the kidneys. Progressively more uremic retention solutes have been identified and their potential toxicity has been characterized. Polypeptides constitute a heterogeneous group of uremic molecules. Therefore, proteome analysis represents a new and promising analytical approach to identify new uremic toxins. Proteomic technologies cover applicability to a broad molecular mass range. For polypeptides >10 kDa classical proteomic techniques, such as two-dimensional gel electrophoresis followed by mass spectrometry, are able to identify uremic polypeptides. In the mass range from approximately 1 to 10 kDa, capillary electrophoresis coupled to mass spectrometry (CE-MS) emerged as a fast possibility to analyze of up to 1,400 compounds in a single step. This chapter will provide an overview about proteomic technologies as efficient tools for the detection of uremic toxins, emphasizing the features of CE-MS. Subsequently, examples of the application of proteomic techniques to define novel biomarkers for renal diseases and uremic toxins will be discussed. PMID- 18401169 TI - Proteomic approaches for the study of cell signaling in the renal collecting duct. AB - In the current era of large-scale biology, proteomics has evolved as a powerful, new technique that aims to identify, quantify, and analyze a large number of proteins in a functional context. Therefore, proteomics can be used to study cellular pathways and identify disease biomarkers. In this review, we first outline the principles of two important proteomics techniques that either use difference gel electrophoresis (DIGE) or liquid chromatography (LC) for protein separation, followed by tandem mass spectrometry (MS/MS). The advantages and limitations of each technique are discussed, emphasizing the ability of DIGE to perform quantitative proteomics and the high-throughput and high-sensitivity characteristics of LC-MS/MS. We have employed both techniques to unravel the molecular machinery of vasopressin signaling, which governs water homeostasis by recruiting aquaporin-2 (AQP2) water channels after activation of the vasopressin 2 receptor by vasopressin. Several aspects of vasopressin signaling in the inner medullary collecting duct (IMCD) were investigated, including the short- and long term regulation of AQP2, phosphoproteomics, signaling during vasopressin escape, and the proteomes of AQP2-bearing vesicles and the IMCD plasma membranes. We also emphasize that proteomics of body fluids will be the strategy to identify disease biomarkers, and therefore conclude the review by highlighting the perspectives of biomarker discovery in urinary exosomes. PMID- 18401170 TI - Overview of kidney and urine proteome databases. AB - With the completion or almost completion of genome sequences of many organisms in combination with the tremendous development of mass spectrometric analysis of proteins, several comprehensive proteomic studies, targeting whole organisms, body fluids, organs, tissues, cells, cellular organelles, or functional protein complexes, have produced valuable resources that can be shared and retrieved. In the present review, we provide current concept of construction of protein databases with special emphasis on high-throughput identification of protein using mass spectrometry, annotations, computational tools, and search engines to retrieve information of the identified proteins. We then update the current status of available protein databases of kidney and urine proteomes. PMID- 18401171 TI - Distribution of APOE genotypes in a memory clinic cohort. AB - AIM: To describe the distribution of apolipoprotein E (APOE) genotypes in a cohort of memory clinic patients. METHODS: We included 749 memory clinic patients. Diagnoses were made in a multidisciplinary consensus meeting and the APOE genotype was determined. The community-based cohort of the Longitudinal Aging Study Amsterdam was used as control population (n = 2,233). RESULTS: In the memory clinic sample, there were 173 patients with subjective complaints, 125 patients with mild cognitive impairment (MCI), 251 patients with Alzheimer disease (AD), 107 patients with another type of dementia, and 93 patients with another neurologic or psychiatric diagnosis. The APOE allele distribution differed among groups. There was no difference in the prevalence of the epsilon2 allele, but there were differences in distribution of the epsilon3 and epsilon4 alleles. Compared with the control population (15%), the prevalence of APOE epsilon4 was increased among patients with subjective complaints (22%), MCI (36%), AD (42%) and other types of dementia (25%). CONCLUSION: We observed an increased prevalence of APOE epsilon4 in patients with MCI and subjective complaints. This finding is of great clinical importance as nondemented patients positive for APOE epsilon4 could be identified as being at genetic risk of AD, and for that reason monitored more closely. PMID- 18401172 TI - Cognitive function and psychiatric symptoms in early- and late-onset frontotemporal dementia. AB - BACKGROUND/AIM: Some recent studies mentioned that late-onset frontotemporal dementia (FTD) is more common than previously assumed. Although much research has been done in the field, there are no systematic studies which have compared clinical characteristics of early- and late-onset FTD. The aim of this study was to compare cognitive function and psychiatric symptoms in patients with early- and late-onset FTD. METHODS: Study participants were consecutive outpatients. There were 35 FTD patients; their mean age at onset was 63.0 years. We studied sex, education, duration from onset to consultation, Clinical Dementia Rating (CDR) scores, Mini-Mental State Examination (MMSE) scores, Raven's Coloured Progressive Matrices (RCPM) scores, and Neuropsychiatric Inventory (NPI) scores at first consultation of early- and late-onset FTD patients. RESULTS: There were no significant differences in sex ratio, education, CDR scores, and duration from onset to consultation. There were significant differences in the total MMSE scores, 'three-word recall task', 'construction task', and RCPM scores; late onset groups scored significantly lower than early-onset groups. There were significant differences in the apathy domain of NPI and total NPI scores; late onset groups scored significantly higher than early-onset groups. CONCLUSION: Late-onset FTD patients may have memory and visuospatial deficits in addition to their behavioural changes, even if they are clinically diagnosed according to consensus diagnostic criteria. They also present more apathy, and they may have a different histolopathological background. PMID- 18401173 TI - Effect of ApoE genotype on response to donepezil in patients with Alzheimer's disease. AB - BACKGROUND/AIMS: The possible influence of apolipoprotein E (ApoE) genotype on the response to acetylcholinesterase inhibitor therapy in patients with Alzheimer's disease (AD) remains a matter of controversy. In order to address this issue, we investigated the effects of ApoE genotype on the clinical response to donepezil in patients with mild to moderate AD. METHODS: An open study was carried out in 51 patients with probable AD who were treated with 5-10 mg of donepezil per day for 48 weeks. RESULTS: Eighteen (35.3%) of the 51 patients had 1 or 2 ApoE epsilon4 alleles. ApoE epsilon4 carriers with AD showed a mean 1.1 point increase from the baseline score of 23.9 on the 70-point Alzheimer's Disease Assessment Scale-Cognitive Component at 48 weeks, while the ApoE epsilon4 noncarrier group showed a 3.1-point increase from the baseline score of 22.5 (p = 0.03). The ApoE epsilon4 carrier group exhibited a mean 0.13-point worsening from the baseline score of 0.97 on the Korean Instrumental Activities of Daily Living at 48 weeks, while the ApoE epsilon4 noncarrier group exhibited a 0.17-point worsening from the baseline score of 0.64 (p = 0.05). CONCLUSION: AD patients who carry the ApoE epsilon4 allele may respond more favorably to donepezil than epsilon4 noncarriers. PMID- 18401174 TI - Vascular cognitive impairment: prodromal stages of ischemic vascular dementia. AB - BACKGROUND/AIMS: To describe the natural history of the prodromal stages of ischemic vascular dementia (pVaD). METHODS: A sample of 314 inpatients with pVaD or a clini- cal diagnosis of vascular dementia (VaD; lacunar state, Binswanger's disease, pure cortical VaD, corticosubcortical and strategic infarctions) admitted to a teaching tertiary center during a 13-year period was assessed (retrospectively n = 88, prospectively n = 226). Prospective neuropsychological assessment consisted of Mini Mental State Examination, Revised Wechsler Adult Intelligence Scale, Exit-25, Trail Making tests, Blessed Dementia Scale and Camdex H, Global Depression Scale and Hamilton Depression Rating Scale tests. Univariate analysis and logistic regressions are displayed. RESULTS: An unrecognized pVaD was related with a clinical onset with cognitive impairment no dementia (CIND) versus symptomatic cerebrovascular events (p < 0.0001), and with being under therapy with anticoagulant or antiplatelet agents (p < 0.01). Age <85 years at diagnosis of VaD (p < 0.01) correlated with a delayed pVaD diagnosis. CIND onset was associated with a longer prodromal stage (p < 0.01), no clinical strokes during pVaD (p < 0.001), silent ischemia (p < 0.01) and Binswanger's disease (p < 0.01). CONCLUSIONS: Vascular cognitive impairment remains an underdiagnosed, yet treatable entity. A brief neuropsychological examination and informant interviews should become standard practice in elderly populations with vascular risk factors. Small-vessel disease is a prevalent condition with a distinct natural history. PMID- 18401175 TI - Lentigo maligna in a young adult. AB - BACKGROUND: Lentigo maligna is mainly diagnosed in the elderly with severely sun damaged skin. OBSERVATION: We describe a case of lentigo maligna in a 51-year-old woman with Fitzpatrick skin type III. Based on private photographs of the patient, we were able to ascertain that the lesion was already present at the age of 22 years. Despite the 29-year clinical history, the lesion was still entirely intraepidermal on histologic examination. CONCLUSION: Lentigo maligna in young adults is extremely rare, and therefore represents a diagnostic challenge for both clinicians and histopathologists. PMID- 18401176 TI - Papillon-Lefevre syndrome and malignant melanoma. A high incidence of melanoma development in Japanese palmoplantar keratoderma patients. AB - Papillon-Lefevre syndrome (PLS) is a rare autosomal-recessive genodermatosis characterized by palmoplantar hyperkeratosis and severe early-onset periodontitis. The development of malignant cutaneous neoplasms within the hyperkeratotic lesions of the syndrome is quite rare. Here, we report on a 51 year-old Japanese woman with PLS associated with recurrent malignant melanoma (MM). Mutation analysis of the cathepsin C gene revealed that the proband was homozygous for a missense mutation, c.415G-->A, which is predicted to result in the amino acid substitution p.G139R. Including our case, 4 families have been described as having PLS with MM, 3 of which are Japanese, implying a high incidence of melanoma development in Japanese PLS patients. We suggest that hereditary palmoplantar keratoderma (PPK) in Japanese patients might be predisposed to MM. A literature review revealed that in 18 cases of MM-associated PPK, 13 (76%) were Japanese, suggesting a high incidence of MM in Japanese PPK patients. This tendency might be attributable to the high frequency of acral lentiginous melanoma in Japanese subjects, in contrast to a lower frequency of this subtype in Caucasians. PMID- 18401177 TI - Acquired unilateral melanocytic nevi in otherwise normal skin. AB - We describe a case with numerous melanocytic nevi in otherwise normal skin. A 5 year-old girl presented with more than 100 small pigment lesions on her left arm, shoulder and upper back without underlying light brown macule. The pigment lesions were first found on her left forearm at 3 months old and gradually increased along with her growth. Skin biopsy from a pigmented lesion shows a pathological change in compound-type melanocytic nevus without any atypical changes. Speckled lentiginous nevus is known to have multiple melanocytic lesions on the underlying brown macule from birth. Partial unilateral lentiginosis is characterized by unilateral lentigines with histopathological changes in lentigo but not melanocytic proliferation in the dermis. Agminated melanocytic nevi tend to be clustered together in a circumscribed area, whereas in the present case melanocytic nevi were segmentally arranged but not agminated. We consider that this is an unusual type of mosaicism of melanocytic disorders. PMID- 18401178 TI - Morphological analysis of the interstitial ultrasonic ablation in porcine liver in vivo. AB - BACKGROUND: The shape of the induced thermal ablation area is as important as its dimension. The aim of this study was to analyze the size reproducibility and the aspect of the interstitial ultrasonic ablation obtained by a planar transducer in porcine liver in vivo. METHODS: Five pigs were used. Two complete ultrasonic lesions were made in each animal under pedicle clamping. All the lesions underwent MR examination on day 7 and then a histological analysis. RESULTS: The tested probe has the advantage of providing a step-by-step and highly directional treatment in the target zone. The ultrasonic lesions presented as well-defined and homogenous areas of tissue coagulation. The lesion volumes ranged from 8.1 to 92.3 cm3 with an averaged lesion length of 56 mm at gross examination. Three dimensional reconstruction of the lesions from the MR images showed cylindrical and conical shapes. Large intrahepatic vessels distorted the lesion shape, and the vicinity of the application to the liver surface increased significantly the volume of the ultrasonic necrosis. Histological examination showed complete necrosis in the area of damage. CONCLUSION: The ultrasonic ablation has a regular shape, always with sharply defined borders. However, it showed some variability in the size of the induced lesions. PMID- 18401179 TI - Preserved BK channel function in vasospastic myocytes from a dog model of subarachnoid hemorrhage. AB - Cerebral vasospasm after subarachnoid hemorrhage (SAH) is due to contraction of smooth muscle cells in the cerebral arteries. The mechanism of this contraction, however, is not well understood. Smooth muscle contraction is regulated in part by membrane potential, which is determined by K+ conductance in smooth muscle. Voltage-gated (Kv) and large-conductance, Ca2+-activated K+ (BK) channels dominate arterial smooth muscle K+ conductance. Vasospastic smooth muscle cells are depolarized relative to normal cells, but whether this is due to altered Kv or BK channel function has not been determined. This study determined if BK channels are altered during vasospasm after SAH in dogs. We first characterized BK channels in basilar-artery smooth muscle using whole-cell patch clamping and single-channel recordings. Next, we compared BK channel function between normal and vasospastic cells. There were no significant differences between normal and vasospastic cells in BK current density, kinetics, Ca2+ and voltage sensitivity, single-channel conductance or apparent Ca2+ affinity. Basilar-artery myocytes had no, small- or intermediate-conductance, Ca2+-activated K+ channels. The lack of difference in BK channels between vasospastic and control cells suggests alteration(s) in other K+ channels or other ionic conductances may underlie the membrane depolarization and vasoconstriction observed during vasospasm after SAH. PMID- 18401180 TI - Does the endothelium abolish or promote arterial vasomotion in rat mesenteric arteries? Explanations for the seemingly contradictory effects. AB - BACKGROUND AND AIMS: Vasomotion consists in cyclic oscillations of the arterial diameter induced by the synchronized activity of the smooth muscle cells. So far, contradictory results have emerged in the literature about the role of the endothelium in the onset and maintenance of vasomotion. Here our aim is to understand how the endothelium may either abolish or promote vasomotion. METHODS AND RESULTS: We investigate rat mesenteric arterial strips stimulated with phenylephrine (PE). Our results show that the endothelium is not necessary for vasomotion. However, when the endothelium is removed, the PE concentration needed to induce vasomotion is lower and the rhythmic contractions occur for a narrower range of PE concentrations. We demonstrate that endothelium-derived relaxing products may either induce or abolish vasomotion. On the one hand, when the strip is tonically contracted in a nonoscillating state, an endothelium-derived relaxation may induce vasomotion. On the other hand, if the strip displays vasomotion with a medium mean contraction, a relaxation may induce a transition to a nonoscillating state with a small contraction. CONCLUSION: Our findings clarify the role of the endothelium on vasomotion and reconcile the seemingly contradictory observations reported in the literature. PMID- 18401182 TI - Endometrial adenocarcinoma: prevention and early diagnosis. PMID- 18401181 TI - The circulating inactive form of matrix Gla Protein (ucMGP) as a biomarker for cardiovascular calcification. AB - OBJECTIVE: Matrix gamma-carboxyglutamate (Gla) protein (MGP) is a vitamin K dependent protein and a strong inhibitor of vascular calcification. Vitamin K deficiency leads to inactive uncarboxylated MGP (ucMGP), which accumulates at sites of arterial calcification. We hypothesized that as a result of ucMGP deposition around arterial calcification, the circulating fraction of ucMGP is decreased. Here we report on the development of an ucMGP assay and the potential diagnostic utility of monitoring serum ucMGP levels. METHODS AND RESULTS: An ELISA-based assay was developed with which circulating ucMGP can be determined. Serum ucMGP levels were measured in healthy subjects (n = 165) and in four patient populations; patients who underwent angioplasty (n = 30), patients with aortic stenosis (n = 25), hemodialysis patients (n = 52), and calciphylaxis patients (n = 10). All four patient populations had significantly lower ucMGP levels. In angioplasty patients and in those with aortic stenosis, some overlap was observed with the control population. However, in the hemodialysis and calciphylaxis populations, virtually all subjects had ucMGP levels below the normal adult range. CONCLUSION: Serum ucMGP may be used as a biomarker to identify those at risk for developing vascular calcification. This assay may become an important tool in the diagnosis of cardiovascular calcification. PMID- 18401192 TI - Evaluation of factors associated with chronic low back pain in hemodialysis patients. AB - BACKGROUND/AIMS: Low back pain is a common and disabling symptom that has not been properly studied in hemodialysis patients. The aim of this study was to evaluate factors associated with chronic low back pain in these patients. METHODS: We evaluated 205 patients undergoing chronic hemodialysis. Data were obtained through medical records, patient interviews and physical examination. Balance was evaluated through the Tinetti scale. The muscular strength was evaluated through the quadriceps muscle manual test and the Kendall scale graduation. RESULTS: 74 patients (36.1%) had low back pain. They were older (p = 0.020), undergoing dialysis for a longer period (p = 0.071) and had more comorbidities than patients without lumbar pain. Among the patients with low back pain, 75.7% presented balance disorders and 59.5% muscular weakness. In multivariate analysis the factors related to low back pain were balance deficit (OR: 9.30; 95% CI: 2.77-31.20), muscular strength deficit (OR: 14.33; 95% CI: 4.55-45.19), arterial hypertension (OR: 4.51; 95% CI: 1.32-15.37), bone disease (OR: 43.39; 95% CI: 11.51-163.53) and cerebrovascular disease (OR: 20.21; 95% CI: 3.28-124.65). CONCLUSIONS: Chronic low back pain is common in hemodialysis patients and is associated with muscle weakness, balance disorders, and comorbidities. PMID- 18401193 TI - Depression on dialysis. AB - Depression is the most common psychopathological condition among patients with end-stage renal disease (ESRD), yet it is still under-recognized and misdiagnosed. Depression reduces quality of life and has a negative clinical impact upon sufferers with chronic illness, including ESRD. This article discusses the negative effects of depression among the ESRD population treated with dialysis, the prevalence of the condition, the methodological issues involved with screening and treatment, and the possible psychological and somatic causes. There is a need to identify the prevalence of the disorder by effective methods, overcome the current issues surrounding depression assessment and to undertake trials of suitable treatments. PMID- 18401195 TI - Transcutaneous electrical stimulation of subjective tinnitus. A placebo controlled, randomized and comparative analysis. AB - AIM: The effect of the transcutaneous application of the electrical stimulus on tinnitus perception has been reviewed in a placebo-controlled, randomized and comparative analysis to eventually determine the outcome of the therapeutic role of the therapy. METHOD: There are 42 patients who were randomized into 2 groups according to their order of admission. Group A consists of 31 patients who were subjected to transcutaneous electrical stimulation 3 times a week for 1 month. Group B includes 11 patients who had electrical stimulus attachment but where no stimulus was given (placebo group). The stimulator is a custom-made device which generates direct and alternative current in 10-200 Hz frequency. An alternative low-frequency (not >100 Hz) pulsed current was used for tinnitus therapy through a preauricular skin electrode. The amplitude of stimulus ranged between 50 and 2,000 mA. The pulse frequency was 30 Hz. Each session lasted for 25 min for both groups. Statistical analysis was performed. RESULT: The rate of improvement following the therapy was 42.8% (18/42) in the electrical therapy group and 28.5% (4/14) in the placebo group. CONCLUSION: Electrical suppression of the tinnitus does not offer a promising outcome for patients with tinnitus in the presented study. PMID- 18401194 TI - Role of histamine H3 and H4 receptors in mechanical hyperalgesia following peripheral nerve injury. AB - OBJECTIVE: Histamine is a chemical mediator that acts at four known types of histamine receptors and has been widely implicated in the development of nociception and neuropathic pain. Blocking histamine H(1) and H(2) receptors has been shown to reduce hyperalgesia following nerve injury, but the role of histamine H(3) and H(4) receptors in neuropathic pain has not been studied. Here, we used blockers of histamine H(3) and H(4) receptors to assess their effects on neuropathic pain behavior and mast cell numbers following peripheral nerve injury. In addition, we assessed the effect of activating H(4) receptors on neuropathic pain behavior. METHODS: Rats were subjected to a partial ligation of the sciatic nerve, a model of neuropathic pain, and were treated either systemically or locally (hindpaw) with the H(3)/H(4) receptor inverse agonist thioperamide, the specific H(4) receptor antagonist JNJ 7777120, or the H(4) receptor agonist VUF 8430. Measurements of mechanical hyperalgesia were carried out by Randall-Selitto test for 1-3 weeks, and sciatic nerve tissues were analyzed for numbers of intact mast cells by histology at 9 h after surgery. RESULTS: Rats treated with thioperamide or JNJ 7777120 showed significantly enhanced mechanical hyperalgesia after partial ligation of the sciatic nerve. The number of intact mast cells in the injured nerve of these rats was higher than in control rats suggesting reduced mast cell degranulation, but was still significantly lower than in intact nerves. Rats treated with VUF 8430 showed significantly reduced mechanical hyperalgesia. CONCLUSION: We propose that the increase in mechanical hyperalgesia produced by thioperamide and JNJ 7777120 and the decrease in mechanical hyperalgesia produced by VUF 8430 may represent a direct effect of these agents on mechanospecific primary afferents, or an indirect effect of these agents via injury-induced inflammation. PMID- 18401196 TI - Expression of glucose transporter-1 and -3 in the head and neck carcinoma--the correlation of the expression with the biological behaviors. AB - OBJECTIVE: The aim of this study was to determine the biological significance of glucose transporter (Glut)-1 and Glut-3 expression in head and neck carcinoma (HNC). METHODS: We detected expression of Glut-1 and -3 in 38 HNCs and analyzed the relationship between increased expression and the biological behavior of HNCs. RESULTS: The gene expression levels of Glut-1 and -3 in HNCs were significantly higher than those in adjacent cancer tissues or in normal tissues. The GLUT-1 gene level was correlated with the lymph node metastasis and clinical stage of 38 HNCs, and the GLUT-3 gene level was correlated with the lymph node metastasis of 38 HNCs. Of the 38 cases, 30 showed positive expression of Glut-1 protein. The Glut-1 protein expression level was related to the lymph node metastasis and clinical stage of 38 HNCs. However, there was no case that expressed Glut-3 protein. The high expression of Glut-1 gene and protein was associated with poor survival in the HNCs. GLUT-3 gene expression was not associated with the prognosis of HNCs. CONCLUSIONS: GLUT-1 gene expression level and protein expression were correlated with lymph node metastasis, poor survival and clinical stage of HNCs. The GLUT-3 gene expression level was high in HNCs, and its expression was associated with an increased incidence of lymph node metastasis of HNCs. PMID- 18401197 TI - Tinnitus and neurovascular compression. PMID- 18401198 TI - Hyperbaric oxygen treatment restores sudden hearing loss in a patient with Fabry disease. AB - Fabry disease is an X-linked inherited disorder of glycosphingolipid metabolism due to the deficient activity of a lysosomal enzyme, alpha-galactosidase A. The resultant systemic accumulation of sphingolipids can lead to progressive and sudden hearing loss alongside renal, cardiac and cerebrovascular complications. Although replacement therapy seems to be beneficial for cochlear function, few data are available regarding treatment of sudden hearing loss. This case report describes the course of a unilateral sudden hearing loss in a young (15-year-old) male patient and its improvement following hyperbaric oxygen treatment. PMID- 18401199 TI - Intensive insulin therapy in preschool-aged diabetic children: from multiple daily injections to continuous subcutaneous insulin infusion through indwelling catheters. AB - In this study, glycemic control, diabetes care indices and quality of life (QoL) were assessed in 2 groups of newly diagnosed Type 1 diabetic subjects <6 yr old who were randomized to multiple daily injections with (Group A) or without (Group B) an indwelling catheter. Group A [12 males (M)/8 females (F), mean age 3.2+/ 1.4 yr] and Group B (9M/11F, mean age 3.9+/-1.8 yr) were evaluated at baseline and after 6 and 12 months of treatment. No significant difference was observed in metabolic control (glycosylated hemoglobin) or in the number of hypoglycemic events between the groups. Patients in Group A had a greater number of daily insulin injections, monitored blood glucose more frequently and had a lower total daily insulin dose per kg (p<0.05). QoL was better in group A. At the end of the study 30% of group A patients progressed to continuous sc insulin infusion (CSII), while no child in Group B switched to a different insulin regimen. Based on these findings, indwelling catheter therapy may be helpful for selected CSII candidates. PMID- 18401200 TI - Relative cardiac expression of growth hormone receptor and insulin-like growth factor-I mRNA in congenital heart disease. AB - GH may exert direct growth-promoting and metabolic actions on target tissues, but most of its effects are mediated by circulating (endocrine) or local (auto /paracrine) IGF-I. The GH/IGF-I system has an important role in cardiac development and in maintaining the structure and function of the heart. A subgroup of children with pronounced heart defects will eventually need transplants, owing to congestive heart failure. Since the symptoms are often severe and may progress while waiting for surgery, it is necessary to develop supportive medical treatment. GH has been proposed as a therapeutic agent in adults with heart failure, but to date studies are lacking on children and more information is necessary. We have examined the expression of IGF-I mRNA and GH receptor (GH-R) mRNA in children undergoing surgery for congenital heart disease. Eighteen children scheduled for open-heart surgery were included in the study. Right auricular biopsies were taken at the time of venous catheterization preceding cardiac bypass. The specimens were analysed using realtime PCR. We were able to show expression of both IGF-I mRNA and GH-R mRNA in the pediatric heart. The relative expressions were intercorrelated (r=0.75, p<0.001). GH-R mRNA correlated positively to standardized weight (r=0.65, p=0.004), body mass index (BMI) (r=0.59, p=0.01), and standardized BMI (r=0.59, p=0.01). IGF-I mRNA only correlated to BMI (r=0.50, p=0.04). This is the first study displaying cardiac expression of IGF-I mRNA and GH-R mRNA in children with congenital heart disease, although further studies are needed to define a role for GH in the treatment of these patients. PMID- 18401202 TI - Frequency of atopy and allergic disorders among adults with Type 1 diabetes mellitus in the southern Marmara region of Turkey. AB - AIM: Autoimmune disorders are considered to be associated with a Th1 immune response whereas allergic diseases with a Th2 response. Studies mainly performed on children revealed conflicting results regarding the association of atopy/allergic disease and autoimmune disorders. Therefore, we aimed to investigate the prevalence of allergic diseases in adult Type 1 diabetic patients. METHODS: Eighty-nine Type 1 diabetic patients and 64 controls were enrolled into the study. Skin-prick test and European Community Respiratory Health Survey questionnaire were performed on all cases. Patients who gave at least one positive answer to questions about asthma in the questionnaire underwent pulmonary function test and methacholine challenge test. RESULTS: Patients' mean age were similar in diabetic patients and controls (28.2+/-8.9 and 28.1+/-5.2 yr; respectively). In skin-prick test, the rate of positive response to at least one allergen was not significantly different in diabetes (29.2%) and in the control group (31.3%). In European Community Respiratory Health Survey questionnaire, diabetic patients waked up by an attack of cough more than controls did. The rate of physician-diagnosed asthma was similar in both groups. There was no difference between the 2 groups based on the answers of other questions about asthma and other allergic diseases such as allergic rhinitis, eczema, and drug allergy. CONCLUSION: We found that atopy frequencies were similar in an adult population of Type 1 diabetic patients and controls. Although asthmatic symptom prevalence is increased in diabetic patients, the incidence of current asthma was similar in both groups. PMID- 18401201 TI - Cloning of Russian sturgeon (Acipenser gueldenstaedtii) growth hormone and insulin-like growth factor I and their expression in male and female fish during the first period of growth. AB - In this study, the GH and IGF-I of the Russian sturgeon (rs), Acipenser gueldenstaedtii, were cloned and sequenced, and their mRNA gene expression determined. In addition, to improve our understanding of the GH function, the expression of this hormone was assessed in young males and females. Moreover, IGF I expression was quantified in young males and compared to that in older ones. The nucleotide sequence of the rsGH cDNA was 980 bp long and had an open reading frame of 642 bp, beginning with the first ATG codon at position 39 and ending with the stop codon at position 683. A putative polyadenylation signal, AATAAA, was recognized 42 bp upstream of the poly (A) tail. The position of the signal- peptide cleavage site was predicted to be at position 111, yielding a signal peptide of 24 amino-acids (aa) and a mature peptide of 190 aa. When the rsGH aa sequence was compared with other species, the highest degree of identity was found to be with mammalians (66-70% identity), followed by anguilliformes and amphibia (61%) and other fish (39-47%). The level of rsGH mRNA was discovered to be similar in pituitaries of females and males of 5 age groups (1, 2, 3, 4, and 5 yr-old). In females and males, the levels did not change dramatically during the first 5 yr of growth. The partial nucleotide sequence of the rsIGF-I was 445 bp long and had an open reading frame of 396 bp, beginning with the ATG codon at position 50. The position of the signal-peptide cleavage site was predicted to be at position 187, yielding a signal peptide of 44 aa. The highest level of IGF-I mRNA expression was recorded in the kidney of adult sturgeons. The IGF-I mRNA expression levels in the intestine, pituitary gland, and liver were not significantly different. Low levels of expression were found in the brain, heart, and muscle. In most tissues, there was no significant difference between mRNA levels of one and 5-yr-old fish. In conclusion, based on the GH-sequence analysis, A. gueldenstaedtii is genetically distant from other teleosts. The expression of the GH mRNA was similar in males and females, and its level remained constant during the first 5 yr of growth. While the IGF-I mRNA expression differed amongst various tissues, the level in each tissue was similar in 1 and 5-yr-old fish. PMID- 18401203 TI - Diagnostic and prognostic implications of the World Health Organization classification of neuroendocrine tumors. AB - BACKGROUND: Neuroendocrine differentiation of tumors is often difficult to establish. In the same manner, the evaluation of the prognostic role of neuroendocrine differentiation may constitute a relevant clinical problem. Although different classifications are used for neuroendocrine tumors (NET) of different origin, the last World Health Organization (WHO) classification of NET, originally proposed for gastroenteropancreatic tumors, has proved to be a practical tool to allow pathologists to uniform the diagnoses and re-classify these tumors into 3 main categories. AIM: The present study was carried out in order to evaluate diagnostic and prognostic implications of NET reclassification according to the last WHO classification of NET. MATERIALS AND METHODS: Thirty one tumors with an initial diagnosis referable to a NET achieved before 1999 were independently evaluated by 3 pathologists on the basis of the 2000 WHO classification of NET. Immunohistochemistry for panneuroendocrine markers and Ki 67 was also performed in all cases. RESULTS: Twelve, 14, and 4 tumors were respectively reclassified as well-differentiated NET, well-differentiated neuroendocrine carcinoma and poorly differentiated neuroendocrine carcinoma; 1 tumor was reclassified as mixed endocrine-exocrine tumor. Two or more neuroendocrine markers were expressed in all NET regardless of histotype, differentiation degree, and site of primary tumor. After revision, 10 of the 31 tumors under study (32%) changed histo-prognostic category when compared to the initial diagnosis. Ki-67 score was the best predictor of survival at the multivariate analysis. CONCLUSION: The WHO classification is suitable to accurately reclassify tumors with an initial diagnosis referable to a NET and to separate these tumors in 3 well-distinct histo-prognostic categories with relevant clinical implications. Ki-67 score seems to be a better predictor of survival than the degree of differentiation. PMID- 18401204 TI - Acute ghrelin response to intravenous dexamethasone administration in idiopathic short stature or isolated idiopathic growth hormone-deficient children. AB - Acylated ghrelin has been originally described for its potent GH-releasing activity mediated by the activation of the GH secretagogue receptor type 1a. More recently, ghrelin has been reported to exert several other GH-independent biological actions, among which in the modulation of metabolic functions. Glucocorticoids are well known to exert important metabolic functions but also to modulate GH secretion, although through mechanisms that have not been fully clarified so far. Interestingly, the existence of a feedback link between glucocorticoids and ghrelin system has already been reported. The aim of our study was to evaluate the acute GH and ghrelin responses to dexamethasone (DEX) administration in children with idiopathic short stature (ISS) or isolated idiopathic GH deficiency (GHD). Eight children with ISS (age: 9.5+/-1.2 yr) and 7 with GHD (12.1+/-1.4 yr) underwent iv DEX administration (0.3 mg/body surface area at 0 min). IGF-I, GH, and ghrelin levels were assayed at baseline and every 30 min from 120 up to 240 min after DEX. Compared to baseline levels DEX decreased ghrelin in ISS at 120 min and 240 min (p<0.04). On the other hand DEX did not modify ghrelin levels in GHD. After DEX, ghrelin was reduced in ISS compared to GHD (p<0.02). DEX increased GH in ISS but not in GHD (peak: 11.1+/ 1.2 vs 7.6+/-0.9 microg/l). Basal, as well as after-DEX ghrelin levels negatively correlated with IGF-I in GHD (p<0.03) and with height SD score (HSDS) in ISS (p<0.02). Acute DEX administration is able to decrease ghrelin in ISS, but not in GHD children. Both basal and after-DEX ghrelin levels negatively correlate with IGF-I and HSDS. All these data suggest the existence of a feedback link among ghrelin, glucocorticoids and the GH/IGF-I axis. PMID- 18401205 TI - Direct effects of microgravity on testicular function: analysis of hystological, molecular and physiologic parameters. AB - Spaceflight experiments carried out in microgravity environments have revealed that exposure to altered gravity condition results in alteration of several cellular functions and, consequently, of several apparatuses. There is some evidence in the literature indicating that spaceflight affects the physiology of the testis. The data on effects of spaceflight or simulated microgravity on testicular function, however, sometimes appear contradictory. In the present study we used an in vitro experimental model in order to investigate the direct effects of microgravity on testicular tissue. We generated a microgravity environment using the Rotating Wall Vessel and performed experiments on testicular fragments isolated from pre-pubertal rats. In this model we then analyzed several parameters such as histological integrity, cell proliferation, cell apoptosis, occludin distribution pattern, and hormonal secretions. The emerging picture shows some alterations of testicular tissue physiology. Interestingly, we also demonstrate for the first time that, in organ culture, Leydig cell survival is severely affected by simulated microgravity. PMID- 18401206 TI - Glucocorticoid effect on insulin sensitivity: a time frame. AB - It is well known that glucocorticoids induce insulin resistance, but the exact time scale in humans is not well known. The aim of the study was to determine the time scale of effects of pharmacologic doses of glucocorticoids on insulin sensitivity. Subjects were treated with repeated methylprednisolone infusions and oral prednisone for Graves' orbitopathy. Insulin sensitivity was determined using euglycemic hyperinsulinemic clamp (EHGC) before, during the first glucocorticoid infusion and after 2 months of treatment. EHGC started 2 h after the start of the glucocorticoid infusion, and lasted for 2 h. In another group of patients, insulin sensitivity was determined by short insulin tolerance test (SITT) before and during the first glucocorticoid infusion. SITT started 15 min after the start of the glucocorticoid infusion and lasted for 15 min. Ten subjects were included in each protocol. All were euthyroid during the study period. Four hours after the start of the glucocorticoid infusion significant reduction of insulin sensitivity was observed, which did not change for a further 2 months of glucocorticoid treatment [before 7.82 (95% confidence interval (CI) 5.35-10.29), first infusion, 4.93 (95% CI 2.99-6.87), after 2 months 5.36 (95%CI 3.91-6.81) mg/kg/min]. No significant change in insulin sensitivity occurred during the first 30 min of glucocorticoid infusion [before 139.7 (95%CI 94.1-185.3), during 146.7 (95%CI 106.3-187.1) mumol/l/min]. In humans, glucocorticoid- induced insulin resistance develops quickly, in about 4 h, and does not change during further glucocorticoid treatment. PMID- 18401207 TI - A five-year follow-up study of goiter and thyroid nodules in three regions with different iodine intakes in China. AB - OBJECTIVE: The association between iodine status and the prevalence of goiter and thyroid nodules has been well established but the extent to which different iodine intake levels influence the incidence of goiter and thyroid nodules is unclear. The aim of the study was to determine the incidence of goiter and thyroid nodules in 3 regions with different iodine intake levels: mildly deficient, more than adequate, and excessive. DESIGN, PATIENTS AND MEASUREMENTS: Of the 3385 unselected subjects enrolled in 1999 in Panshan, Zhangwu, and Huanghua where median urinary iodine excretion (UIE) was 83.5 microg/l, 242.9 microg/l, and 650.9 microg/l, respectively, 2708 (80.0%) participated in the follow-up study in 2004. The examinations of thyroid ultrasonography, thyroid function, thyroid autoantibodies and UIE were performed at baseline and follow up. RESULTS: The cumulative incidence of diffuse goiter was 7.1%, 4.4%, and 6.9%, respectively, higher in Panshan and Huanghua than in Zhangwu (p=0.013 and p=0.015) and that of nodular goiter was 5.0%, 2.4%, and 0.8%, respectively, declining with increasing iodine intake levels (p<0.001). Mild iodine deficiency, chronic iodine excess as well as positive thyroid autoantibodies were associated with the occurrence of goiter [Logistic regression: odds ratio (OR)=1.83 (95% confidence interval (CI) 1.26-2.65), OR=1.46 (95% CI 1.01-2.11) and OR=1.68 (95% CI 1.14-2.48), respectively]. The cumulative incidence of single nodule was 4.0%, 5.7%, and 5.6%, respectively and that of multiple nodules was 0.4%, 1.2%, and 1.0%, respectively. CONCLUSIONS: The relationship between iodine and the risk for the occurrence of diffuse goiter shows a U-shaped curve. Nodular goiters are more prevalent in iodine-deficient areas. PMID- 18401208 TI - Effect of inappropriate and continuous therapy with alendronate for ten years on skeletal integrity - observations in two elderly patients. AB - Alendronate is a potent aminobisphosphonate that has been used worldwide to decrease fracture risk in millions of post-menopausal women with and without osteoporosis, men with low bone mass, and in those with glucocorticoid- induced osteoporosis. A recent report of 9 patients with spontaneous atypical non vertebral fractures during treatment with alendronate for up to 8 yr raised questions suggesting the possibility of severe suppression of bone turnover and resultant susceptibility to fracture. Our recent observations in 2 elderly women with inactive monostotic Paget's disease of bone who had been treated elsewhere continuously for this disease with alendronate for 10 yr at doses overall of 2 and 4 times the osteoporotic dose provided an opportunity to engage in the ongoing controversy over long-term safety of bisphosphonate therapy. Despite such therapy, skeletal integrity was maintained with normal bone densities and the absence of skeletal fractures. These observations do not support the suggestion of deleterious effects with longterm alendronate therapy. PMID- 18401210 TI - Acute keratoconus induced by hypothyroxinemia during pregnancy. AB - Keratoconus is a bilateral non-inflammatory corneal ectasia with an incidence of approximately 1 per 2000 in the general population. The etiology of keratoconus is multifactorial, although not completely understood. Several clinical observations suggest that thyroid gland dysfunction is associated with keratoconus pathophysiology. Pregnancy represents a major alteration in the maternal endocrinologic status characterized by modified function of the thyroid gland. We report on a 33-yr-old pregnant women suffering from keratoconus, who presented simultaneously with a remarkable deterioration of the disease during gestation and with thyroid gland dysfunction. Acute keratoconus in this patients coincided with the lowest plasma T4 level reached during pregnancy. During the third trimester of gestation, the keratoconus receded significantly. Here we provide direct evidence in support of a possible linkage between keratoconus and thyroid gland dysfunction, assumed by previous clinical observations. This case report is the first to suggest that changes in maternal thyroid hormone levels during pregnancy can aggravate the progression of keratoconus. PMID- 18401209 TI - Clinical and biochemical improvement in acromegaly during pregnancy. AB - Numerous case reports of pregnancy in acromegaly exist, however detailed descriptions of changes in placental and pituitary GH and IGF-I throughout gestation are rare. A 19-yr-old female presented to this institution with signs and symptoms of a GH-secreting pituitary adenoma. Following transphenoidal hypophysectomy, she had 3 unplanned pregnancies, despite ongoing active disease. No pregnancy was complicated by glucose intolerance or hypertension and 3 healthy newborns were delivered near or at term. Clinical improvement was observed during each pregnancy, accompanied by IGF-I levels lower than in the non-pregnant state, the majority lying within the normal range. This was despite increasing placental GH levels, and was not consistent with previous reports in the literature. Further surgical and medical therapies for acromegaly failed to normalize nonpregnant GH or IGF-I levels in this woman. Estrogen is known to alter GH signaling via its interaction with Janus kinase/signal transducers and activators of transcription (JAK-STAT) pathways. We hypothesize that increasing concentrations of estrogen or other pregnancy-related hormones resulted in her clinical and biochemical improvement during pregnancy. This may be used for future therapeutic benefit. PMID- 18401211 TI - Skeletal involvement in adult patients with endogenous hypercortisolism. AB - Overt endogenous glucocorticoid excess is a well-recognized cause of bone loss and osteoporotic fractures. Cortisol excess inhibits bone formation, increases bone resorption, impairs calcium absorption from the gut, and affects the secretion of several hormones (in particular gonadotropins and GH), cytokines, and growth factors, influencing bone metabolism. The glucocorticoid excess mainly affects trabecular bone, leading to vertebral fractures in up to 70% of patients. Osteoporotic fractures may be the presenting symptom of an otherwise silent glucocorticoid excess and can precede the diagnosis of hypercortisolism by up to 2 yr. The removal of glucocorticoid excess leads to a recovery of bone mass which is, however, often incomplete and delayed, although it reduces the risk of osteoporotic fractures. Bisphosphonate therapy has been suggested to be useful in maintaining bone mass in these patients. Subclinical hypercortisolism, a condition of impaired hypothalamic- adrenal-axis homeostasis without the classical signs and symptoms of glucocorticoid excess, is a recently defined entity, which has been shown to be associated to increased bone resorption, bone loss, and high prevalence of vertebral fractures regardless of gonadal status. However, data about the effect of this subtle glucocorticoid excess on bone are still scarce and conflicting. Moreover, it is not yet known whether removing the cause of subclinical hypercortisolism leads to a recovery of bone mass and reduces the risk of osteoporotic fractures. Finally, recent data suggest that subclinical hypercortisolism is a common and underrated finding in patients with established osteoporosis. In summary, it is crucial to evaluate the risk of osteoporosis and fractures in patients with glucocorticoid excess; on the other hand, it also seems advisable to screen for glucocorticoid excess patients with osteoporotic fractures without known secondary causes of osteoporosis. PMID- 18401213 TI - Transcranial Doppler blood flow assessment in patients with mild heart failure: correlates with neuroimaging and cognitive performance. AB - Cardiac output and cerebral perfusion are reduced in patients with advanced stages of heart failure. Our aim was to determine whether cerebral blood flow velocity measured by transcranial Doppler ultrasonography was reduced in outpatients with mild heart failure in comparison to controls and, if so, whether this reduction was related to cognitive performance and abnormalities of the brain diagnosed by magnetic resonance imaging. PMID- 18401212 TI - The biological characterization of neuroendocrine tumors: the role of neuroendocrine markers. AB - Neuroendocrine tumors (NET) may originate in different organs, from cells embryologically different but expressing common phenotypic characteristics, such as: the immuno-reactivity for markers of neuroendocrine differentiation (defined as "pan-neuroendocrine"), the capacity to secrete specific or aspecific peptide and hormones and the expression of some receptors, that are at the basis of the current diagnostic and therapeutical approach, peculiar to these tumors. NET have been conventionally distinguished in functioning, when associated with a recognized clinical endocrine syndrome, and non-functioning. However, this terminology may be misleading, since the great majority of NET may secrete neuroendocrine peptides, which can be employed as clinical markers for both diagnosis and follow-up. On the other hand, tissue immuno-reactivity for specific hormones does not always reflect secretory activity of the tumor cells. Finally, receptors and genetic markers are acquiring a relevant role in the characterization of NET, both improving knowledge of biology and physiopathology of NET, as well as in developing specific strategies to establish an early diagnosis and targeted therapies, to adopt prophylactic strategies in familial forms, and to identify more efficacious targets for therapy in the future. PMID- 18401214 TI - Baseline characteristics of patients randomized in The Resynchronization Reverses Remodeling In Systolic Left Ventricular Dysfunction (REVERSE) study. AB - The Resynchronization Reverses Remodeling in Systolic Left Ventricular Dysfunction (REVERSE) study is a randomized controlled trial currently assessing the safety and efficacy of cardiac resynchronization therapy in patients with asymptomatic left ventricular (LV) dysfunction with previous symptoms of mild heart failure. This paper describes the baseline characteristics of randomized patients; 610 patients with New York Heart Association (NYHA) class II (82.3%) heart failure or asymptomatic (NYHA class I) LV dysfunction with previous symptoms (17.7%) were randomized in 73 centers. The mean age was 62.5+/-11.0 years, the mean LV ejection fraction was 26.7%+/-7.0%, and the mean LV end diastolic diameter was 66.9+/-8.9 mm. A total of 97% of patients were taking angiotensin-converting enzyme inhibitors or angiotensin receptor blockers and 95.1% were taking beta-blockers, which were at the target dose in 35.1% of patients. Compared with previous randomized cardiac resynchronization therapy trials, REVERSE patients are on better pharmacologic treatment, are younger, and have a narrower QRS width despite similar LV dysfunction. PMID- 18401215 TI - Nonsteroidal anti-inflammatory drugs and the heart: what is the danger? AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) and selective cyclooxygenase (COX 2) inhibitors, or "coxibs," are used for a number of disease conditions for relief of pain and inflammation. Currently available data suggest concern for prothrombotic risk with coxibs and some NSAIDs, and the magnitude of risk may vary with individual agents. NSAIDs and coxibs also increase blood pressure, worsen hypertension control, and may precipitate heart failure, with important differences among agents. Physicians should consider patterns of risk and benefit in selecting the most appropriate agent for individual patients based on the individual gastrointestinal and cardiovascular risk profile. PMID- 18401216 TI - A practical guide for ultrafiltration in acute decompensated heart failure. AB - Acute decompensated heart failure is the most common reason for inpatient hospital admission. Most patients admitted for decompensated heart failure are by definition diuretic-resistant. The therapeutic objective for these patients is volume and sodium removal and restoration of diuretic sensitivity. In a significant proportion of patients, this objective is not met, subjecting patients to readmission for recurrent heart failure decompensation. Ultrafiltration therapy offers the potential of greater volume and sodium removal as compared with conventional therapies in a more expeditious manner. Ultrafiltration can be safely and effectively accomplished in a non-intensive care setting but relies on earlier discharge with reduced readmission rates to be economically feasible. This paper reviews the current data regarding ultrafiltration therapy and provides a practical guide to patient selection, implementation and management of this therapy. PMID- 18401217 TI - Cardiomyopathy associated with ephedra-containing nutritional supplements. PMID- 18401218 TI - Utility of cardiac magnetic resonance imaging in evaluating left ventricular pseudoaneurysm. PMID- 18401219 TI - Growing hearts by the bushel. PMID- 18401220 TI - Genetic testing and genetic counseling in cardiovascular genetic medicine: overview and preliminary recommendations. AB - In this emerging era of cardiovascular genetic medicine, increasing responsibility will be placed on cardiovascular practitioners to be aware of the latest clinical genetic testing methods and the knowledge base needed to interpret genetic test results. Some cardiovascular specialists will develop the expertise within the field to order genetic testing and interpret results, while other practitioners will refer patients to centers of excellence in cardiovascular genetic medicine. A previous article in the Cardiovascular Genetic Medicine: Clinical Perspectives and Future Applications series(1) highlighted an increasing recognition of the cardiomyopathies (hypertrophic [HCM], dilated [DCM], arrhythmogenic right ventricular dysplasia [ARVD]) and channelopathies (long QT syndrome [LQTS] and others) as genetic diseases, and focused on the importance of a targeted family history as a critical part of patient evaluation. The goal of this article, second in the series, is to provide a general framework for understanding the principles of genetic testing and genetic counseling. We review the growing number of genetic tests currently available to cardiac specialists, the selection of an appropriate test, and the numerous genetic counseling issues raised by the testing process. We also provide our preliminary recommendations for genetic testing in cardiovascular genetic medicine. PMID- 18401221 TI - Cardiomyopathy and multiple pulmonary emboli in a patient with myotonic dystrophy. PMID- 18401222 TI - Pay (adequately) for what works: the economic undervaluation of office and ambulatory blood pressure recordings. PMID- 18401223 TI - A cluster randomized trial to evaluate physician/pharmacist collaboration to improve blood pressure control. AB - This was a prospective, cluster randomized controlled trial in patients with uncontrolled hypertension aged 21 to 85 years (mean, 61 years). Pharmacists made recommendations to physicians for patients in the intervention clinics (n=101) but not patients in the control clinics (n=78). The mean adjusted difference in systolic blood pressure (BP) between the control and intervention groups was 8.7 mm Hg (95% confidence interval [CI], 4.4-12.9), while the difference in diastolic BP was 5.4 mm Hg (CI, 2.8-8.0) at 9 months. The 24-hour BP levels showed similar effects, with a mean systolic BP level that was 8.8 mm Hg lower (CI, 5.0-12.6) and a mean diastolic BP level that was 4.6 mm Hg (CI, 2.4-6.8) lower in the intervention group. BP was controlled in 89.1% of patients in the intervention group and 52.9% in the control group (adjusted odds ratio, 8.9; CI, 3.8-20.7; P<.001). Physician/pharmacist collaboration achieved significantly better mean BP values and overall BP control rates, primarily by intensification of medication therapy and improving patient adherence. PMID- 18401224 TI - The use of olmesartan medoxomil as monotherapy or in combination with other antihypertensive agents in elderly hypertensive patients in Japan. AB - The efficacy and safety of the angiotensin receptor blocker olmesartan medoxomil (OLM) was assessed in 550 elderly Japanese hypertensive patients who were followed for 24 weeks in daily clinical practice. Patients were given OLM alone or in combination with other antihypertensive drugs at the discretion of the investigators. After 24 weeks of treatment, systolic and diastolic blood pressure (BP) significantly decreased from baseline (P<.0001). When patients were classified as either young-old (65-74 years) or older-old (75 years and older), with either isolated systolic hypertension (ISH) or systolic-diastolic hypertension (SDH), the reduction of diastolic BP in ISH patients was significantly smaller than that in SDH patients (5.0 vs 15.2 mm Hg; P<.0001), indicating that OLM did not cause excessive reduction of diastolic BP in ISH patients. Treatment was well tolerated in all groups. In conclusion, the medication was safe and effective in reducing BP levels in ISH patients aged 75 years and older, as well as in other elderly hypertensive patients. PMID- 18401225 TI - Use of home blood pressure monitoring by hypertensive patients in primary care: survey of a practice-based research network cohort. AB - Using a survey of a cohort of primary care patients, the authors determined the proportion currently using home blood pressure monitoring (HBPM) and calculated odds ratios (ORs) of factors associated with such use. Overall, 530 questionnaires were received (80% response rate); 35.2% of respondents reported that their doctor had recommended HBPM (95% confidence interval [CI], 31.1-39.3), and 43.1% reported currently using HBPM (95% CI, 38.8-47.3). Compared with patients younger than 45 years, hypertensive patients older than 65 years were more likely to be using HBPM (OR, 2.53; 95% CI, 1.20-5.33). Those with a history of stroke/transient ischemic attack were also more likely to use HBPM (OR, 2.06; 95% CI, 1.00-4.24). Compared with patients with a level of hypertension knowledge <10th percentile, those with a knowledge level >90th percentile were more likely to use HBPM (OR, 1.96; 95% CI, 1.08-3.56). The factor most strongly associated with use of HBPM was recalling a doctor's recommendation to do so (OR, 7.93; 95% CI, 4.96-12.7). PMID- 18401226 TI - Perceived and actual stroke risk among men with hypertension. AB - The purposes of this study were to determine whether there is a significant correlation between the perceived and actual stroke risk among hypertensive patients and to identify patient characteristics associated with inaccurate estimation of stroke risk. The authors performed a cross-sectional analysis of 296 men with hypertension who were enrolled in the Veterans Study to Improve the Control of Hypertension (V-STITCH). A patient's actual stroke risk was calculated using the Framingham stroke risk (FSR); patients' perceived risk was measured according to a self-reported 10-point risk scale. The median 10-year FSR was 16%, but the median perceived risk score was 5 (range, 1 [lowest] to 10 [highest]). There was no significant correlation between patients' perceived risk of stroke and their calculated FSR (Spearman rho=-0.08; P=.16; 95% confidence interval, 0.19 to 0.03). Patients who underestimated their stroke risk were significantly less likely to be worried about their blood pressure than patients with accurate risk perception (12.4% vs 69.6%; P<.0001). The lack of correlation between hypertensive patients' perceived stroke risk and FSR supports the need for better patient education on the risks associated with hypertension. PMID- 18401227 TI - Effects of arterial stiffness, pulse wave velocity, and wave reflections on the central aortic pressure waveform. AB - Brachial systolic and pulse blood pressures (BPs) are better predictors of adverse cardiovascular (CV) events than diastolic BP in individuals older than 50 years. The principal cause of increased systolic and pulse BP is increased stiffness of the elastic arteries as a result of degeneration and hyperplasia of the arterial wall. Recent studies have shown that central BP, the pressure exerted on the heart, brain, and kidneys, is a better predictor of CV risk than brachial BP. As stiffness increases, reflected wave amplitude increases and augments pressure in late systole, producing an increase in left ventricular afterload and myocardial oxygen demand. Vasoactive drugs have little direct effect on large human elastic arteries but can markedly modify wave reflection by altering stiffness of the muscular arteries and changing pulse wave velocity of the reflected wave from the periphery to the heart. Vasodilators decrease the amplitude and increase the travel time (or delay) of the reflected wave, causing a generalized decrease in systolic BP. The decrease in systolic BP brought about by this mechanism is grossly underestimated when systolic BP is measured in the brachial artery. PMID- 18401228 TI - Vascular system: role of nitric oxide in cardiovascular diseases. AB - In contrast with the short research history of the enzymatic synthesis of nitric oxide (NO), the introduction of nitrate-containing compounds for medicinal purposes marked its 150th anniversary in 1997. Glyceryl trinitrate (nitroglycerin) is the first compound of this category. On October 12, 1998, the Nobel Assembly awarded the Nobel Prize in Medicine or Physiology to scientists Robert Furchgott, Louis Ignarro, and Ferid Murad for their discoveries concerning NO as a signaling molecule in the cardiovascular system. NO-mediated signaling is a recognized component in various physiologic processes (eg, smooth muscle relaxation, inhibition of platelet and leukocyte aggregation, attenuation of vascular smooth muscle cell proliferation, neurotransmission, and immune defense), to name only a few. NO has also been implicated in the pathology of many inflammatory diseases, including arthritis, myocarditis, colitis, and nephritis and a large number of pathologic conditions such as amyotrophic lateral sclerosis, cancer, diabetes, and neurodegenerative diseases. Some of these processes (eg, smooth muscle relaxation, platelet aggregation, and neurotransmission) require only a brief production of NO at low nanomolar concentrations and are dependent on the recruitment of cyclic guanosine monophosphate (cGMP)-dependent signaling. Other processes are associated with direct interaction of NO or reactive nitrogen species derived from it with target proteins and requires a more sustained production of NO at higher concentrations but do not involve the cGMP pathway. PMID- 18401229 TI - Implementing effective hypertension quality improvement strategies: barriers and potential solutions. AB - Many quality improvement strategies have focused on improving blood pressure control, and these strategies can target the patient, the provider, and/or the system. Strategies that seem to have the biggest effect on blood pressure outcomes are team change, patient education, facilitated relay of clinical information, and promotion of self-management. Barriers to effective blood pressure control can affect the patient, the physician, the system, and/or "cues to action."We review the barriers to achieving blood pressure control and describe current and potential creative strategies for optimizing blood pressure control. These include home-based disease management, combined patient and provider education, and automatic decision support systems. Future research must address which components of quality improvement interventions are most successful in achieving blood pressure control. PMID- 18401230 TI - Well tolerated spironolactone-related hyponatremia. PMID- 18401231 TI - What is the best drug to prescribe for a young woman in her childbearing years with essential hypertension? PMID- 18401232 TI - Global risk, women, men: what's the score? PMID- 18401233 TI - Efficacy of aerobic exercise on coronary heart disease risk factors. AB - The authors examined the effects of aerobic exercise on selected coronary heart disease (CHD) risk factors using data from previously published meta-analyses. Using a random effects model, the effects of aerobic exercise on glycosylated hemoglobin (HbA1c) (mean, 95% confidence interval, -0.9%, -1.9% to 0.03%), resting systolic blood pressure (-6.9 mm Hg, -9.1 to -4.6 mm Hg), low-density lipoprotein cholesterol (-3.1 mg/dL, -6.1 to 0 mg/dL), and body mass index (-1.3 kg/m(2), -2.5 to -0.1 kg/m(2)) were either statistically significant or demonstrated a trend for statistical significance. Changes were equivalent to relative reductions of -8.5%, -4.7%, -2.0%, and -4.5%, respectively. Changes corresponded to estimated 5-year reductions in CHD mortality of 14%, 17%, 1.5%, and 5%, respectively. The results of this review reinforce the idea that aerobic exercise is an important nonpharmacologic intervention for improving selected CHD risk factors. PMID- 18401234 TI - Cardiovascular risk factors among internal medicine residents. AB - Modification of risk factors can reduce the number of deaths due to cardiovascular disease (CVD). Internal medicine (IM) residents devote significant clinical time to help patients modify CVD risk factors but may fail to recognize the presence of such factors in their own lives. The prevalence of major modifiable risk factors was assessed in IM residents. Of 101 eligible residents, 56 completed at least 1 component of the study. None had symptoms or history of CVD, and 11 (20%) had > or = 2 CVD risk factors. Of 39 subjects who had physical assessment, 3 had systolic hypertension and 13 had a body mass index > or = 25. Of 38 patients with fasting lipid measurements, 13 had total cholesterol >200 mg/dL; 28 had low-density lipoprotein > or = 100 mg/dL, with 7 >160 mg/dL; and 7 had high-density lipoprotein <40 mg/dL. Three residents smoked 10 cigarettes per day and 15 were sedentary. Training in IM should include strategies to increase awareness of modifiable personal risk factors for CVD, as well as strategies to reduce or eliminate them. PMID- 18401235 TI - The effect of tai chi exercise on blood pressure: a systematic review. AB - A systematic review of the literature on the effect of tai chi exercise on blood pressure (BP) was performed. The authors searched Medline, CAB, Alt HealthWatch, BIOSIS previews, Science Citation Index, and EMBASE systems (inception through January 2007); researched Chinese Medical, China Hospital Knowledge, China National Knowledge Infrastructure, and China Traditional Chinese Medicine databases (inception to June 2005); and performed hand searches at the medical libraries of Beijing and Nanjing Universities. Clinical studies of tai chi examining BP as an outcome published in English or Chinese were included. Studies reporting only acute exercise effects were excluded. Data were extracted in a standardized manner and 2 independent investigators assessed methodologic quality. Twenty-six studies examining patients with and without cardiovascular conditions met inclusion criteria: 9 randomized controlled trials, 13 nonrandomized studies, and 4 observational studies. Study heterogeneity precluded formal meta-analyses. Twenty-two studies (85%) reported reductions in BP with tai chi (3-32 mm Hg systolic and 2-18 mm Hg diastolic BP reductions). Five randomized controlled trials were of adequate quality (Jadad score > or = 3). No adverse effects were reported. Tai chi exercise may reduce BP and serve as a practical, nonpharmacologic adjunct to conventional hypertension management. PMID- 18401236 TI - The accuracy of aneroid sphygmomanometers in the ambulatory setting. AB - The mercury sphygmomanometer is the undisputed gold standard for the indirect measurement of blood pressure. Some public health advocates have recently expressed concern about the use of mercury in medical practice.(2) This concern has prompted many medical facilities to replace mercury manometers with aneroid devices. The present report examined the performance of 282 aneroid sphygmomanometers in outpatient medical practices. Results were examined for predetermined end points within +/-3 mm Hg from the reference values and to indicate zero at no pressure. Ninety-three devices (33%) failed to perform at > or = 1 pressure levels. Most (76%) of the failures were due to low readings. Only 7 of the 93 failing units did not rest at zero, making this an unreliable indicator of accuracy. Inaccurate readings of aneroid sphygmomanometers may result in a failure to diagnose and treat hypertension, thereby placing hypertensive patients at risk for end-organ damage and cardiovascular events. PMID- 18401237 TI - Elevated levels of homocysteine predict cardiovascular death, nonfatal myocardial infarction, and symptomatic bypass graft disease at 2-year follow-up following coronary artery bypass surgery. AB - Severe graft disease occurs in patients at a rate of approximately 15% within the first year of coronary artery bypass grafting (CABG). In this study, the authors examined predictors of the combined end point of death, nonfatal myocardial infarction (MI), and bypass graft disease at 2-year follow-up after CABG. One hundred twenty-one consecutive patients were included in this study after informed consent was obtained. In univariate analysis, there was a significantly (P<.05) higher homocysteine level (11.0 ng/mol vs 9.7 ng/mol, P=.04) in patients who met the combined end point vs those who did not. There were no statistically significant differences in the following: low-density lipoprotein cholesterol, high-sensitivity C-reactive protein, and lipoprotein(a) values; age; body mass index; smoking and diabetes status; statin or aspirin use; creatinine level; hematologic markers; left ventricular ejection fraction; number of bypass grafts; and distribution of coronary artery disease. Logistic regression analysis modeling for low-density lipoprotein cholesterol, lipoprotein(a), fibrinogen, and homocysteine showed that homocysteine value (P=.016) was an independent predictor of the primary combined end point. PMID- 18401238 TI - Peak oxygen consumption and the minute ventilation/carbon dioxide production relation slope in morbidly obese men and women: influence of subject effort and body mass index. AB - The authors evaluated the minute ventilation/carbon dioxide production relation (VE/VCO2 slope) as a complementary measure to peak oxygen consumption (peak VO2) in 76 patients (mean +/- SD age = 44.3+/-10.8 years, 69.7% female) with morbid obesity (mean +/- SD body mass index [BMI] = 49.4+/-7.0 kg/m(2)), as it is not limited by effort. Nearly one-half (43%) of the patients achieved a peak respiratory exchange ratio <1.10. Mean peak VO2 and VE/VCO2 slope were 17.0+/-3.7 mL/kg/min and 27.8+/-4.0, respectively. Peak VO2 correlated with BMI (r=-0.45, P<.0001), while VE/VCO2 slope did not (r=-0.04, P=.73). There was a linear trend for declining mean peak VO2 (P=.001) but not for VE /VCO2 slope (P=.59) with increasing BMI quintiles. The VE/VCO2 slope is an effort-independent measure that is also independent of BMI and may serve as an adjunctive cardiorespiratory variable when evaluating morbidly obese men and women. PMID- 18401239 TI - Postpartum adiponectin concentration, insulin resistance and metabolic abnormalities among women with pregnancy-induced disturbances. AB - The authors compared postpartum adiponectin levels among women with prior pregnancy-induced disturbances and assessed their association with homeostasis model assessment for insulin resistance (HOMA-IR), the metabolic syndrome (MS), and the Framingham risk score (FRS). Women delivering in 1998 through 2001 and who had gestational diabetes mellitus (n=22), gestational hypertension (n=32), or preeclampsia (n=34) were examined 1 to 2 years after delivery and were grouped matched to controls (n=29) by age and prepregnancy body mass index. HOMA-IR was increased, adiponectin values were decreased, and there was a higher MS prevalence in women with prior gestational diabetes mellitus (all P<.05). Adiponectin levels were inversely related to HOMA-IR (r=-0.45; P<.0001) and FRS (r=-0.25; P=.007), and a significant trend for decreasing adiponectin values with increased number of MS components was noted (P trend <.0001). Adiponectin concentration remained a significant correlate of FRS and MS irrespective of pregnancy history; a concentration <10.5 microg/mL provided the optimal cutoff to distinguish those with or without MS. Thus, a lower postpartum adiponectin concentration identifies women at increased cardiovascular risk regardless of pregnancy history. PMID- 18401240 TI - Clinical characteristics and outcomes in hypertensive patients of Hispanic descent. AB - In the United States, patients of Hispanic descent have higher rates of hypertension-related morbidity and mortality than patients from other ethnic groups even though the prevalence of hypertension among Hispanics is lower. This discrepancy likely reflects lower rates of hypertension awareness and control among Hispanics as well as a higher prevalence of diabetes mellitus and other cardiovascular risk factors in this population. Although available data suggest that patients of Hispanic descent who receive antihypertensive therapy experience responses and treatment benefits similar to those achieved by other ethnic groups, clinical trials of antihypertensive therapy have typically enrolled only small numbers of Hispanic patients. Agents targeting the renin-angiotensin system, namely angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers, may be particularly useful in the Hispanic population given the ability of these drugs to protect against hypertension-related and diabetes related end-organ damage independent of their blood pressure-lowering effects. PMID- 18401241 TI - Portion distortion. PMID- 18401243 TI - American Society for Preventive Cardiology. PMID- 18401242 TI - ILLUMINATE-Part 2 and CORONA. PMID- 18401254 TI - Use of sedatives, opioids, and neuromuscular blocking agents in patients with acute lung injury and acute respiratory distress syndrome. AB - OBJECTIVE: The use of sedatives, opioids, and neuromuscular blocking agents (NMBAs) may delay weaning and prolong intensive care unit length of stay. We hypothesized that in patients on higher positive end-expiratory pressure (PEEP), sedatives, opioids, and NMBAs are used in a higher proportion of patients and in higher doses and that the use of these medications is associated with prolongation of weaning and mortality. DESIGN: Retrospective analysis. SETTING: The ALVEOLI trial. PATIENTS: Five hundred forty-nine patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS) who were enrolled in the ALVEOLI trial. INTERVENTIONS: We analyzed prospectively collected data regarding the impact of sedatives, opioids, and NMBAs in ALI/ARDS patients on duration of mechanical ventilation, time to weaning landmarks, and mortality. MEASUREMENTS AND MAIN RESULTS: Sedatives and opioids were used in >80% of the patients in similar proportion in the two groups. The use of sedatives and opioids, but not the use of NMBAs, was associated with longer time on mechanical ventilation and an increased time to achieve a 2-hr spontaneous breathing trial (p < .0001). Sedatives were also associated with increased time to achieve unassisted breathing. NMBAs were used for a short period of time, in a higher proportion of patients in the lower PEEP group, and for a longer time (0.23 days). CONCLUSIONS: Sedatives and opioids use was similar in the higher and lower PEEP groups. The use of sedatives and opioids, but not NMBAs, was associated with a longer time to achieve important weaning landmarks. PMID- 18401255 TI - Pulse contour cardiac output analysis in a piglet model of severe hemorrhagic shock. AB - OBJECTIVE: Pulse contour cardiac output (PCCO) analysis is a technique for continuous cardiac output (CO) monitoring through an arterial catheter after calibration by transpulmonary thermodilution (TPTD). Studies in adults show good correlation with pulmonary artery thermodilution (PATD) CO. Data are limited in children and patients with hemodynamic instability. The objective was to determine whether TPTD CO and PCCO analysis correlate with PATD CO in a piglet model of severe hemorrhagic shock. Mixed venous oxygen saturation (SVO2) was also compared with PATD CO. DESIGN: Prospective animal study. SETTING: University animal research laboratory. SUBJECTS: Domesticated piglets, 24-37 kg. INTERVENTIONS: Hemorrhagic shock was created by graded hemorrhage in anesthetized piglets. Hemorrhage was initiated to achieve mean arterial pressure plateaus of 60, 50, 40, 30, and 20 mm Hg. MEASUREMENTS AND MAIN RESULTS: CO was measured by PATD and simultaneously with two femoral artery PCCO catheters. At each mean arterial pressure plateau, one PCCO catheter was recalibrated by TPTD; the other catheter was not recalibrated during hemorrhage. TPTD CO, PCCO measurements from each catheter, and SVO2 were compared with PATD CO at each mean arterial pressure level. TPTD CO and recalibrated PCCO showed excellent correlation (r2 = .96 and .97) and small bias (+0.11 and +0.14 L/min), respectively, compared with PATD. Without recalibration, PCCO measurements were not accurate during rapid hemorrhage (r2 = .22). SVO2 decline did not correlate as well with PATD CO (r2 = .69). CONCLUSIONS: TPTD CO and recalibrated PCCO analysis correlate well with PATD CO in this severe hemorrhagic shock model. The mean difference is small (<0.15 L/min) and is not clinically significant. With rapid changes in blood pressure or intravascular volume, PCCO is not accurate unless recalibrated by TPTD CO. SVO2 did not correlate well with CO in this model. PMID- 18401256 TI - Introduction: targets for improving outcomes in renal allografts transplanted during the next ten years. PMID- 18401257 TI - Vasculopathy in the kidney allograft at time of transplantation: impact on later function of the graft. AB - BACKGROUND: Older and marginal donors, increasingly used to overcome organ shortness, often have a cerebrovascular accident as cause of death and could have vascular lesions in their kidneys. METHODS: In this literature study, we evaluated the predictive value of vasculopathy in the renal allograft at the time of transplantation, on the subsequent graft function. RESULTS: Short-term graft survival rates do not seem to be diminished by suboptimal donor histology. When vasculopathy is clearly present at the time of transplantation, impaired kidney function is showed at 1-week posttransplantation, at hospital discharge, or at 3 months and an increased frequency of delayed graft function. The long-term graft survival rate, in the studies of Pokorna and Taub, was significantly lower in the group with arteriolosclerosis. Wang et al. concluded in their study that arterial fibrous intimal thickening is the single most important histological predictor of both graft loss and delayed graft function. However, Minakawa et al. observed no significant correlation between 1 or 2-year graft survival and vasculopathy score. Severe vascular lesions in the donor kidney do affect the level of kidney function in the later posttransplant period as described in different studies (follow-up till 7 years posttransplantation). CONCLUSIONS: Data obtained from the studies of donor biopsies sustain the notion that vasculopathy is a major determinant of the short-term and the long-term outcome of the kidney allograft. PMID- 18401258 TI - Biotransformation enzymes and drug transporters pharmacogenetics in relation to immunosuppressive drugs: impact on pharmacokinetics and clinical outcome. AB - Immunosuppressive drugs commonly used after organ transplantation to prevent acute rejection including tacrolimus, cyclosporine, sirolimus, and mycophenolic acid are characterized by a narrow therapeutic index and broad interindividual variability in their pharmacokinetics. Adequate immunosuppression aims to reach an optimal benefit-risk ratio. Therapeutic drug monitoring represents a crucial step in routine practice to maintain blood concentrations within the target window, because the bioavailability of these drugs depends on their absorption, distribution, biotransformation, and elimination. Single nucleotide polymorphisms (SNPs) in genes encoding biotransformation enzymes (CYP3A) and drug transporters (ABCB1) have opened up a promising way for the selection of individual dosages. The relationship of these SNPs with immunosuppressive drug pharmacokinetics was extensively studied after kidney, liver, heart, and lung transplantations. Patient susceptibility to nephrotoxicity in the long term was also reported in relation to some SNPs, which could allow effective assessment of individual risk and selection of treatment according to patient parameters. Further studies are needed to provide evidence that a genetic analysis combined with therapeutic drug monitoring has the potential to optimize drug use after transplantation. PMID- 18401259 TI - Immunosuppressive drug therapy and subclinical acute renal allograft rejection: impact and effect. AB - The incidence of subclinical acute rejection (SCAR) varies between 5% and 15% with current maintenance immunosuppressive drug regimens. Despite many similarities between SCAR and clinical acute rejection exist, the inflammatory activated cell infiltrates are not completely identical while graft cytokine profiles and counteractive immune responses are characterized by subtle differences that could explain why SCAR is not accompanied by immediate graft dysfunction. Evidence that SCAR contributes to chronic allograft damage (interstitial fibrosis and tubular atrophy) and negatively affects graft outcome is counterbalanced by the scarcity of controlled data proving the beneficial effect of SCAR treatment. The development of sensitive and specific noninvasive methods to monitor the immune status of the graft by using mRNA determinations, gene expression analysis (microarrays), proteomic analysis, and magnetic resonance spectroscopy, can help to ultimately replace protocol biopsies and also contribute to the further unraveling of the complex underlying immunological mechanisms responsible for SCAR. The latter would enable clinicians to preemptively make strategic adjustments to immunosuppressive therapy in an attempt to further improve renal allograft survival and clinical care of the transplant patient. PMID- 18401260 TI - Cold ischemia is a major determinant of acute rejection and renal graft survival in the modern era of immunosuppression. AB - BACKGROUND: The aim of our study was to examine, in a recent cohort of kidney transplant recipients who have received modern immunosuppressive therapy, the respective role of cold ischemia time (CIT) and delayed graft function (DGF) on acute rejection (AR) rates and long-term graft survival. METHODS: We retrospectively reviewed the charts of 611 renal transplantations between 1996 and 2005. Most patients received a calcineurin inhibitor as maintenance therapy, either cyclosporine (43%) or tacrolimus (52%) and 76% of the patients received an antilymphocyte induction therapy. Study endpoints were DGF, first-year AR, and long-term graft survival. Uni- and multivariate analyses were performed to determine factors that may have influenced the study outcomes. RESULTS: DGF was observed in 16.2% of patients. Both older donor age and longer CIT were significant risk factors for DGF. DGF rates were similar whether patients received a calcineurin inhibitor before transplantation or not. AR occurred in 16.5% of grafts during the first year. Independent predictors of AR by multivariate analysis were duration of dialysis, CIT, current panel-reactive lymphocytotoxic antibody more than 5%, and the number of human leukocyte antigen A, B, and DR mismatches. Each hour of cold ischemia increases the risk of rejection by 4%. With respect to death-censored graft survival, three pretransplant parameters emerged as independent predictors of graft loss: younger recipient age, peak panel-reactive lymphocytotoxic antibody more than 5% and longer CIT. The detrimental effect of CIT on graft survival was entirely because of its propensity to trigger AR. When AR was added to the multivariate Cox model, CIT was no longer significant whereas first-year AR became the most important predictor of graft loss (Hazards ratio, 4.6). CONCLUSION: Shortening CIT will help to decrease not only DGF rates but also AR incidence and hence graft loss. Patients with prolonged CIT should receive adequate immunosuppression, possibly with antilymphocyte preparations, to prevent AR occurrence. PMID- 18401261 TI - Therapeutic interventions favorably influencing delayed and slow graft function in kidney transplantation: mission impossible? AB - Dialyzed delayed graft function (DGF) and nondialyzed slow graft function (SGF) both determine a large portion of the negative impact on patient and graft survival, on early acute rejection incidence, on tubulointerstitial fibrosis/tubular atrophy, and on graft function. In an era where expanded criteria donors, including older donors, and non-heart beating donors are used to overcome organ shortage, the long-term results of kidney transplantation are influenced by the events occurring within the first 24 hr before and after the transplantation procedure. The ischemia-reperfusion injury is initiated in the brain-death donor and continues during preservation and engraftment of the transplant. The determinants of this DGF/SGF complex intervene in five chronologically related stages--donor tissue quality, brain death and related stress, preservation variables, immune factors, and recipient variables. For each of these five stages, therapeutic interventions or preventive measures are capable of partially reversing the expected dismal outcome of DGF/SGF; these measures are discussed in this article. Donor selection and pharmacologic modulations do have an effect on DGF/SGF. PMID- 18401262 TI - Chronic allograft nephropathy: what have we learned from protocol biopsies? AB - Chronic allograft nephropathy, characterized by interstitial fibrosis and tubular atrophy, is still a major cause of graft loss after kidney transplantation. The complex pathophysiology of chronic allograft nephropathy is still poorly understood, and could be clarified by a more systematic performance of implantation and protocol biopsies of the renal allograft. This review highlights the contribution of implantation and protocol biopsies to our current knowledge of the complex interaction of multiple processes, ultimately leading to the development of interstitial fibrosis and tubular atrophy in the transplanted kidney. In addition, the safety and the limitations of protocol biopsies are discussed, as well as potential future directions for clinical practice and clinical research. PMID- 18401263 TI - Polyomavirus in renal transplantation: a hot problem. AB - Polyomavirus BK has emerged as an important complication after kidney transplantation. Although, BK nephropathy develops in only 1% to 5% of renal transplant recipients, its prognosis when present is very poor. The most accepted risk factor is the level of immunosuppressive treatment, but the serostatus of donor and recipient and the absence of human leukocyte antigen C7 in donor and/or recipient influence the BK virus (BKV) reactivation. The gold standard in diagnosing BKV nephropathy (BKVN) continues to be biopsy with use of immunohistochemistry for large T antigens. Urinary decoy cells and blood BKV DNA polymerase chain reaction are used in the screening, but their positive predictive values are poor. However, their use as predictors of the evolution of BKVN is more valuable. The reduction of immunosuppressive therapy currently represents the first-line treatment for BKVN. Cidofovir and leflunomide can be used when BKVN continues to progress. In the event of graft loss, retransplantation is possible with a low risk of recurrence when the infection is no longer active. PMID- 18401264 TI - Do clinical rehabilitation education programs really improve stroke-related knowledge? AB - OBJECTIVE: We evaluated the effectiveness of a clinical stroke education program for improving stroke-related health knowledge after inpatient rehabilitation in a "real world" setting. DESIGN: Thirty-four patients participated in an inpatient rehabilitation clinical stroke education program. Their stroke-related health knowledge in three key domains-risk factors, warning signs, and appropriate actions to take if a stroke is suspected-was evaluated at admission and 12 wks later using a single-group, pretest-posttest design. Pretest and posttest comparisons were conducted using the Marginal Homogeneity test and the McNemar test. RESULTS: Small, nonsignificant improvements in stroke-related health knowledge were detected at posttest. Twelve weeks after the education program, 29% of participants were unable to name a single risk factor, 32% were unable to name a single warning sign, and 29% were unable to name appropriate emergency action in the event they suspected a stroke. CONCLUSIONS: A substantial proportion of patients who completed our clinical stroke education program continued to have poor stroke-related health knowledge. We noted several limitations in our program that may have contributed to this outcome. Changes may be useful for improving the success of clinical stroke education programs, thereby reducing knowledge deficits. PMID- 18401265 TI - Powered mobility for middle-aged and older adults: systematic review of outcomes and appraisal of published evidence. AB - OBJECTIVE: To identify the outcomes of power mobility devices for middle-aged and older adult users, and to critically appraise the research evidence. DESIGN: Systematic review of primary source studies involving adults aged 50 and over using power mobility devices (1996-2007). Articles were (i) mapped to the Taxonomy of Assistive Technology Device Outcomes, which describes categories of impact of assistive devices from the vantages of effectiveness, social significance, and subjective well-being; and (ii) appraised using the Grading of Recommendations, Assessment, Development, and Evaluation criteria. RESULTS: This review retained 19 studies and identified 52 different categories of impacts of power mobility devices spanning the three vantages of the taxonomy. The coverage of outcome dimensions was not as extensive for adults age 50 and over as it was for mixed-age groups. Most of the research designs were assigned very low evidence grades. Three studies were low to moderate in quality of evidence, among which one was a randomized trial. CONCLUSIONS: A vast array of potential impacts of powered mobility devices have been described in the last decade. The level of quality of this evidence is improving, but most of these studies were not designed to verify causal relationships, and this is largely responsible for the absence of unequivocal evidence for directly attributing benefits to devices themselves and for quantifying relationships between power mobility device intervention and outcome. To raise the level of evidence about power mobility device interventions in older adults, studies are needed that use prospective designs, better-defined user groups, and well-grounded conceptual frameworks for measuring interventions and outcomes. PMID- 18401266 TI - Who is performing stapedectomy surgery? Implications for residency and fellowship training. AB - OBJECTIVES: To demonstrate that 1) recent graduates of training programs in otorhinolaryngology-head and neck surgery are less likely to recommend/perform stapedectomy than more senior otolaryngologists; and 2) when surgery is recommended, referral is most commonly made to an otologist/neurotologist. STUDY DESIGN: Survey of 500 regional otolaryngologists pertaining to their treatment of patients with hearing loss secondary to otosclerosis. METHODS: Otolaryngologists in community practice were provided with written surveys pertaining to their treatment of otosclerosis. RESULTS: Data were obtained from 179 general otolaryngologists treating adults and children in solo or group private practices in our geographic region. The majority (66%) diagnosed one to five new cases per year. Ten percent of surgeons graduating in the 1970s, 25% graduating in the 1980s, 50% graduating in the 1990s, and 90% of graduates in the 2000s never performed stapedectomy as part of their practices (P < .001). Similarly, a significant number of surgeons who formerly performed stapedectomies no longer do this surgery. A trend toward greater use of hearing aids for the treatment of otosclerosis was seen in more recent graduates (P < .08). When surgery was recommended, otologists/neurotologists received the majority of referrals (75%) from the practitioners surveyed. CONCLUSIONS: Stapedectomy is performed and recommended less often by more recent graduates of otolaryngology training programs. Training requirements for both general otolaryngologists and neurotologists may need modification given current practice patterns for stapes surgery. PMID- 18401268 TI - Free abdominal fat transfer for reconstruction of the total parotidectomy defect. AB - OBJECTIVES: Free abdominal fat transfer (FAT) grafts have been used after superficial parotidectomy to restore facial contour. We reviewed our experience with the use of FAT reconstruction of total parotidectomy defects to determine the safety and efficacy of this procedure. STUDY DESIGN: Non-randomized retrospective analysis. MATERIALS AND METHODS: The medical records of all patients who underwent total parotidectomy with the harmonic scalpel at the Medical College of Georgia from September 2004 to June 2007 were retrospectively reviewed. Patients with a prior bleeding disorder, facial nerve weakness, or who had undergone concurrent neck dissection were excluded. RESULTS: Twenty-two patients met the study criteria; nine patients underwent FAT reconstruction. There was a greater percentage of malignancy in the FAT group (44%) compared with controls (15%), and a larger tumor size in the FAT group (3.8 +/- 2.1 cm) compared with controls (2.6 +/- 1.2 cm), but these differences did not achieve significance. FAT reconstruction was associated with increased length of surgery (324.6 +/- 86.9 minutes vs. 206.6 +/- 61.8 minutes for controls, P = .0013) and intraoperative blood loss (138.9 +/- 95.3 mL vs. 50.1 +/- 30.9 mL, P = .0052). There were no significant differences between FAT and control groups with respect to postoperative drain output (42.0 +/- 35.0 mL vs. 52.5 +/- 41.3, P > .05), duration of drainage (32.0 +/- 12.0 h vs. 34.8 +/- 15.1 h, P > .05), or facial nerve function. All patients who underwent FAT reconstruction were satisfied with their postoperative appearance, compared to 70% of control patients who reported some degree of dissatisfaction postoperatively. CONCLUSIONS: FAT reconstruction of the total parotidectomy defect is associated with greater blood loss from the FAT donor site and increased surgical time, but is safe and highly effective. These data suggest that the benefits of FAT reconstruction outweigh the additional increment in operative time and blood loss observed. PMID- 18401267 TI - Onset and offset phonation threshold flow in excised canine larynges. AB - OBJECTIVES/HYPOTHESIS: In this study, we examined the hypothesis that the minimum flow required for phonation onset, the onset phonation threshold flow (PTF), is greater than the minimum flow to sustain phonation, the offset PTF. We also explored the hypothesis that the ratio of these two (PTF offset divided by PTF onset) falls within [0.707, 1.0]. STUDY DESIGN: This was a methodology study to measure onset and offset PTFs in 10 excised canine larynges; their ratio (PTF offset divided by PTF onset) was predicted to fall in a specific domain. METHODS: The onset and offset PTF and phonation threshold pressure (PTP) values were observed using 10 excised canine larynges mounted on a bench apparatus. The subglottal flow was increased until phonation was observed, and then decreased until phonation ceased; airflow and pressure measurements at critical conditions of phonation were observed as the PTF and PTP. Larynges with elongated vocal folds were then tested to observe PTF and determine if the hypothesis was observed in pathologic-like larynges. RESULTS: The offset PTF was always less than the onset PTF (P << .0001) and 80.0% of the observed onset-offset PTF ratios were bound by [0.707, 1.0]. CONCLUSIONS: PTF onset or offset could be a useful diagnostic parameter of the voice, particularly when used in conjunction with PTP to describe laryngeal resistance and aerodynamic power. Further exploration of the relationship between onset and offset PTF values could augment clinical diagnostic ability and advance current theories on the physics of phonation. PMID- 18401269 TI - Long-term efficacy of microdebrider-assisted inferior turbinoplasty with lateralization for hypertrophic inferior turbinates in patients with perennial allergic rhinitis. AB - OBJECTIVES: The aim of the study was to evaluate the long-term efficacy of microdebrider-assisted inferior turbinoplasty with lateralization (MAITL) compared to submucosal resection for hypertrophic inferior turbinates. STUDY DESIGN: Surgical outcomes were evaluated with respect to visual analogue scale, anterior rhinomanometry, and saccharin test results. METHODS: From January 2002 to December 2006 inclusively, 160 patients with perennial allergic rhinitis and hypertrophic inferior turbinates were enrolled into this study. The patients, all suffering from chronic nasal obstruction, were randomly classified into two groups, MAITL group or SR group, each comprised of 80 patients. Ten patients who did not display any nasal discomfort served as normal controls. For the submucosal resection group, patients underwent submucosal resection of the inferior turbinate, whereas patients in the MAITL group underwent microdebrider assisted inferior turbinoplasty with lateralization. Assessments (visual analogue scale, anterior rhinomanometry, and saccharin test) were conducted prior to the surgery and 1, 2, and 3 years after completion of surgery. RESULTS: Compared to preoperative values, subjective complaints including nasal obstruction, sneezing, rhinorrhea, and snoring improved significantly in both groups at 1, 2, and 3 years after surgery in both groups (P < .05 for all). Rhinomanometric assessment also showed significant improvement at 1, 2, and 3 years postoperatively in both groups (P < .05 for all). Saccharin transit time was significantly decreased (P < .05 for all) compared to preoperative values 1, 2, and 3 years after surgery in both groups. CONCLUSION: Microdebrider-assisted inferior turbinoplasty with lateralization appears to be as effective as submucosal resection at relieving nasal symptoms and decreasing total nasal resistance and saccharin transit times for more than 3 years in patients with perennial allergic rhinitis who have had substantial nasal obstruction. PMID- 18401270 TI - Desirudin reduces the rate of microvenous thrombosis in a rat model. AB - OBJECTIVES: In an effort to evaluate pharmacologic agents for optimal anticoagulant prophylaxis in patients undergoing free tissue transfer, we evaluated the efficacy of desirudin (Canyon Pharmaceuticals, Hunt Valley, MD), a recombinant hirudin that acts as a direct thrombin inhibitor, using a rat model of microvenous thrombosis. STUDY DESIGN: Randomized, blinded study using an in vivo rat model of microvenous failure. METHODS: Thirty-two rats received either desirudin or saline in a randomized, blinded fashion 30 minutes prior to performance of a standardized thrombogenic procedure on rat femoral veins. Bleeding time, vessel patency, and presence of clot within the anastomosis were subsequently assessed. Appropriate statistical analyses were then performed. RESULTS: There was a significant increase in vessel patency in rats treated preoperatively with desirudin compared to controls receiving saline (96.9% vs. 53.1%, P < .001). In evaluating patent vessels for non-occluding clot, 41.2% of control rats had non-obstructive clot at the site of anastomosis, versus 3.2% of rats treated with desirudin (P = .002). Bleeding times were longer in desirudin treated rats than those that received saline (7.17 +/- 3 minutes vs. 5.15 +/- 1.2 minutes, P = .027). CONCLUSIONS: The use of preoperative desirudin increases the rate of microvascular anastomotic patency, decreases the occurrence of non occluding clot, and increases bleeding time in an in vivo rat model, indicating potential efficacy in patients undergoing microvascular free tissue transfer. PMID- 18401271 TI - Outcomes and efficacy of newborn hearing screening: strengths and weaknesses (success or failure?). AB - OBJECTIVE: To assess the outcomes of neonatal hearing screening with regard to the final diagnosis in a very large number of newborns and investigate related strengths and weaknesses of the program. SUBJECTS: In this study, 76,560 newborns were assessed. METHOD: All neonates were assessed using transient evoked otoacoustic emissions (TEOAEs). RESULTS: From the 76,560 neonates screened, 1,564 (2%) failed the test. According to the screening protocol, all parents of failed neonates were asked to bring their children 1 month following discharge to repeat the test. Of the 541 (34.6%) newborns who repeated the test, 303 (56%) were found normal and 238 (44%) again failed TEOAE. The latter children were referred to two special public centers for full audiology evaluation. In addition, 124 neonates were also referred due to other reasons revealed in the screening process (family history, high levels of bilirubin, etc.). Of the 362 children who were referred to the two special audiology centers, 113 (31.2%) were evaluated by these two centers. In addition, 42 children who had failed initial screening and did not show up for a follow-up appointment to repeat TEOAE were also assessed in the same centers. Of the 155 children who had a special audiologic evaluation, 56 (36.1%) were found to have hearing loss (HL) and 99 (63.9%) normal hearing. In detail, 28 had bilateral sensorineural HL greater than 40 dB, 10 had unilateral sensorineural HL greater than 40 dB, and 18 had otitis media with effusion or other conductive HL. CONCLUSIONS: Derived from the present study: 1) repeated testing of "failed" newborns in the maternity hospital and before discharge leads to an acceptable referral rate of 2%; 2) the 1-month follow-up of "failed" newborns further limits the false positive results but leads to high rate of newborns lost to follow-up; 3) a dedicated secretariat system should be implemented to follow-up each "failed" newborn and remind parents about their follow-up appointments; and 4) additional measures such as detailed educational material and parental friendly approach should also be implemented. PMID- 18401272 TI - Optimization of autologous muscle stem cell survival in the denervated hemilarynx. AB - OBJECTIVE: Current treatments for vocal fold paralysis are suboptimal in that they fail to restore dynamic function. Autologous muscle stem cell (MSC) therapy is a promising potential therapy for vocal fold paralysis in that it can attenuate denervation-induced muscle atrophy and provide a vehicle for delivery of neurotrophic factors, thereby potentially selectively guiding reinnervation. The goal of this project was to characterize optimal conditions for injected autologous MSC survival in the thyroarytenoid (TA) muscle following recurrent laryngeal nerve (RLN) injury by local administration of adjuvant factors. STUDY DESIGN: Animal experiment. METHODS: Unilateral RLN transection and sternocleidomastoid muscle (approximately 1 g) biopsies were performed in 20 male Wistar rats. One month later, 10 autologous MSCs labeled via retroviral-enhanced green fluorescent protein (EGFP) transduction were injected into the denervated hemilarynx of each animal with one of four adjuvant therapies: cardiotoxin [(CTX) 10 M], insulin-like growth factor-1 [(IGF- 1) 100 microg/mL], ciliary neurotrophic factor [(CNTF) 50 microg/mL], or saline. Animals were euthanized 1 month later and larynges harvested, sectioned, and analyzed for MSC survival. RESULTS: All specimens demonstrate extensive MSC survival, with fusion of the MSCs with the denervated myofibers. Based on mean fluorescent intensity of the laryngeal specimens, IGF-1 and CNTF had the greatest positive influence on MSC survival. Myofiber diameters demonstrated myofiber atrophy to be inversely related to MSC survival, with the least atrophy in the groups having the greatest MSC survival. CONCLUSIONS: Autologous MSC therapy may be a future treatment for vocal fold paralysis. These findings support a model whereby MSCs genetically engineered to secrete CNTF and/or IGF-1 may not only promote neural regeneration, but also enhance MSC survival in an autocrine fashion. PMID- 18401273 TI - Evolving attractive faces using morphing technology and a genetic algorithm: a new approach to determining ideal facial aesthetics. AB - OBJECTIVES: The objectives of this study were to: 1) determine if a genetic algorithm in combination with morphing software can be used to evolve more attractive faces; and 2) evaluate whether this approach can be used as a tool to define or identify the attributes of the ideal attractive face. STUDY DESIGN: Basic research study incorporating focus group evaluations. METHODS: Digital images were acquired of 250 female volunteers (18-25 y). Randomly selected images were used to produce a parent generation (P) of 30 synthetic faces using morphing software. Then, a focus group of 17 trained volunteers (18-25 y) scored each face on an attractiveness scale ranging from 1 (unattractive) to 10 (attractive). A genetic algorithm was used to select 30 new pairs from the parent generation, and these were morphed using software to produce a new first generation (F1) of faces. The F1 faces were scored by the focus group, and the process was repeated for a total of four iterations of the algorithm. The algorithm mimics natural selection by using the attractiveness score as the selection pressure; the more attractive faces are more likely to morph. All five generations (P-F4) were then scored by three focus groups: a) surgeons (n = 12), b) cos-metology students (n = 44), and c) undergraduate students (n = 44). Morphometric measurements were made of 33 specific features on each of the 150 synthetic faces, and correlated with attractiveness scores using univariate and multivariate analysis. RESULTS: The average facial attractiveness scores increased with each generation and were 3.66 (+0.60), 4.59 (+/-0.73), 5.50 (+/-0.62), 6.23 (+/-0.31), and 6.39 (+/-0.24) for P and F1-F4 generations, respectively. Histograms of attractiveness score distributions show a significant shift in the skew of each curve toward more attractive faces with each generation. Univariate analysis identified nasal width, eyebrow arch height, and lip thickness as being significantly correlated with attractiveness scores. Multivariate analysis identified a similar collection of morphometric measures. No correlation with more commonly accepted measures such as the length facial thirds or fifths were identified. When images are examined as a montage (by generation), clear distinct trends are identified: oval shaped faces, distinct arched eyebrows, and full lips predominate. Faces evolve to approximate the guidelines suggested by classical canons. F3 and F4 generation faces look profoundly similar. The statistical and qualitative analysis indicates that the algorithm and methodology succeeds in generating successively more attractive faces. CONCLUSIONS: The use of genetic algorithms in combination with a morphing software and traditional focus-group derived attractiveness scores can be used to evolve attractive synthetic faces. We have demonstrated that the evolution of attractive faces can be mimicked in software. Genetic algorithms and morphing provide a robust alternative to traditional approaches rooted in comparing attractiveness scores with a series of morphometric measurements in human subjects. PMID- 18401274 TI - Effects of a novel chitosan gel on mucosal wound healing following endoscopic sinus surgery in a sheep model of chronic rhinosinusitis. AB - INTRODUCTION: Postoperative bleeding and adhesion formation remain the two major problems after endoscopic sinus surgery (ESS). This study investigates the effect on adhesion formation and wound healing in a sheep model of chronic sinusitis of three topical agents: recombinant tissue factor (rTF, Dade Innovin, Marburg, Germany), poly-ethylene glycol (SprayGel, Confluent Surgical, Waltham, MA), and a novel chitosan-dextran derivative gel (CD, Department of Chemistry, University of Otago, Dunedin, New Zealand). METHODS: Twenty sheep with chronic sinusitis underwent ESS with standardized mucosal injuries created on the lateral nasal wall and the ethmoid region. Injured areas were divided into four groups, and one of the three agents or control (no treatment) was randomly applied. The presence and severity of adhesions were noted and the healing was evaluated by taking brushings for ciliary beat frequency and biopsies of the injured regions at day 28, 56, 84, and 112 post initial surgery. The biopsy specimens were assessed for re-epithelialisation using light microscopy and scanning electron microscopy for reciliation. The cytobrush specimens assessed cilial function by measuring ciliary beat frequency. RESULTS: CD significantly decreased lateral nasal wall and ethmoidal adhesions compared to tissue factor at all time points (5% vs. 25%, and 0 vs. 50%, respectively). There was a noticeable trend toward decreased adhesions on the lateral nasal wall and ethmoids in the SprayGel group (10% and 14%) and the CD group (10% and 0%) compared to controls (15% and 40%). The CD group had a significantly greater percentage of re-epithelialisation at day 28 and day 84 compared to the rTF group (70% vs. 33%, P < .001; 84.5% vs. 61%, P < 0.05). At day 28, the CD group was significantly more ciliated than control (62% vs. 31%, P < .01) and than rTF (62% vs. 23%, P < .001). This difference between CD and rTF reciliation remained significant at day 56 (67% vs. 40%, P < .05). In addition, the mean cilial grade for CD at day 112 was significantly better than control (1.9 vs. 2.7, P < .05). CONCLUSION: In the sheep model of chronic sinusitis, CD significantly improves microscopic wound healing and reduces adhesion formation after ESS. PMID- 18401275 TI - Sonographically-directed neck dissection for recurrent thyroid carcinoma. AB - OBJECTIVE: To determine the effectiveness of sonographically-directed compartmental neck dissection for recurrent differentiated thyroid carcinoma as confirmed by ultrasound surveillance and thyroid-specific biochemical markers. STUDY DESIGN: Retrospective chart review. METHODS: Retrospective cohort study of 18 patients with sonographically localized and pathology-confirmed recurrent differentiated thyroid carcinoma. Fifteen patients chose to undergo compartmental neck dissection by a single surgeon. Surgery involved central compartment (level VI) or functional lateral neck dissection (level II-IV+/-V). Three patients declined surgery despite proven recurrent lymph node disease. RESULTS: All 18 patients started out with sonographic evidence of recurrent lymph node disease. Preoperative thyroglobulin or thyroglobulin antibody levels were positive in 17 of 18 patients (94%). Postoperatively, all 15 patients who underwent compartmental lymph node dissections had no sonographic evidence of lymph node disease. Of the 14 patients with preoperative positive thyroglobulin or thyroglobulin antibody levels, 9 patients converted to negative levels (64%). Furthermore, two of four patients (50%) converted to thyroglobulin antibody negative status after surgery. All three patients who declined surgery had persistently detectable sonographic nodal disease in addition to positive thyroglobulin and/or thyroglobulin antibody levels (100%). Surgical complications were minimal and self-limited as no patient experienced permanent cranial nerve deficits. CONCLUSION: Ultrasound-directed cranial nerve sparing compartmental lymph node dissection results in no sonographically detectable cervical lymph node disease and undetectable basal thyroglobulin or thyroglobulin antibody levels in the majority of patients with low morbidity. PMID- 18401276 TI - Immunodefense of the round window. AB - A systematic analysis using serial sectioning of the round window membrane (RWM) in the cynomolgus monkey was performed. Light and transmission electron microscopy (LM and TEM) revealed that the RWM rim may be endowed with gland-like structures with glyco-protein material secernated into the window niche. This was detected in one third of the specimens. The secreted material displayed waste material and scavenger cells. There was also a rich network of capillaries, lymph channels, and sinusoidal veins containing leukocytes. Their abluminal surfaces displayed mature plasma cells and monocytes. These findings suggest that in certain primates the middle ear may have developed specific immunoprotective means for disposal of foreign and noxious substances before they reach the inner ear. PMID- 18401277 TI - Laser disruption of biofilm. AB - OBJECTIVES/HYPOTHESIS: To demonstrate the capability of a fiber-based Q-switched Nd:YAG laser (ARCLaser, Nuremberg, Germany and Valam, Orangeburg, NY) to disrupt biofilm. STUDY DESIGN: Biofilms were grown in broth for 72 hours prior to the experiment. A clinical otorrhea isolate from Pseudomonas aeruginosa was used. Biofilms were placed in MatTek culture plates, on stainless steel screws, tympanostomy tubes, and polyethylene terephthalate (PET) sutures. METHODS: Culture plates, stainless steel screws, tympanostomy tubes, and PET sutures were used for the laser disruption of biofilm experiments. Q-switched Nd:YAG laser pulses were delivered on biofilms using shockwave probes originally designed for cataract surgery. The thin laser fiber tip was targeted against a titanium target, creating the production of plasma and resulting in a shockwave effect. RESULTS: Biofilm areas were imaged before, during, and after laser application using a confocal microscope. The biofilm was imaged growing on the glass/plastic step of the plates, in the grooves of the threads of the screws, over the tympanostomy tube, and on the PET suture. During laser treatment, a time-lapse function was used to capture the results. As a result of laser-generated shockwaves, the biofilm was initially seen to oscillate and eventually break off with individual pulses. Large and small pieces of biofilm were totally and instantly removed from the surface to which they were attached in a matter of a few seconds. CONCLUSIONS: We were able to effectively disrupt Pseudomonas aeruginosa biofilms in vitro using a miniature Q-switched Nd:YAG laser, thin fibers, and special probes that generated plasma formation and a resulting shockwave effect. This laser technology has the ability to generate a powerful stress wave sufficient to disrupt biofilm without any ill effect to the underlying host structure. PMID- 18401278 TI - Flexible endoscopic clip-assisted Zenker's diverticulotomy: the first case series (with videos). AB - BACKGROUND: In treating Zenker's diverticulum (ZD), there are potential risks associated with performing flexible endoscopic diverticulotomy without suturing or stapling. We recently introduced flexible endoscopic clip-assisted diverticulotomy (ECD) in treating ZD by securing the septum prior to dissection. OBJECTIVE: To evaluate the feasibility and safety of ECD for complete septum dissection. STUDY DESIGN: Case series at an academic center. Seven consecutive patients (mean age 71 y; range 48-91 y) with symptomatic ZD of various craniocaudal sizes based on radiographic measurements (mean 2.6 cm; range 0.8 cm 4.5 cm) were included. The mean depth of the septum was 1.73 cm (range 0.3 cm-3.1 cm). The mean duration of symptoms was 4.8 years (range 0.5-10 y). METHODS: After endoclips were placed on either side of the cricopharyngeal bar, the septum was dissected between these two clips down to the inferior end of the diverticulum with a needle-knife. Procedures including "one-step ECD" (n = 1), "stepwise ECD" (n = 3), and "bottom ECD" (n = 2) were performed based on the septum depth of the ZD during endoscopy. ECD was not performed on one patient due to severe mucosal fragility of the esophageal inlet. Iatrogenic blunt dissection of the septum by the endoscopic hood occurred secondary to patient retching during the procedure. Main outcome measurements were symptom resolution and complications. RESULTS: All patients (n = 6) who underwent ECD had complete resolution of esophageal symptoms at a minimum 6-month follow-up. There were no procedural complications. The patient who did not undergo ECD developed an esophageal perforation. She was managed conservatively without surgical intervention. On follow-up, her dysphagia was completely resolved. CONCLUSIONS: ECD is feasible, safe, and effective for complete septum dissection. ECD and endoscopic stapler-assisted diverticulotomy are complimentary rather than competing strategies in approaching ZD. Study limitations include the case series design and limited follow-up period. PMID- 18401279 TI - Clinical validation of percutaneous cochlear implant surgery: initial report. AB - OBJECTIVE: Percutaneous cochlear implant surgery consists of a single drill path from the lateral mastoid cortex to the cochlea via the facial recess. We sought to clinically validate this technique in patients undergoing traditional cochlear implant surgery. STUDY DESIGN: Prospective clinical trial. METHODS: After institutional regulatory board-approved protocols, five ears were studied via the following steps. 1) In the clinic under local anesthesia, bone-implanted anchors were placed surrounding each mastoid. 2) Temporal-bone computed tomography (CT) scans were obtained. 3) On the CT scans, paths were planned from the lateral mastoid cortex, through the facial recess, to the basal turn of the cochlea both "manually" and "automatically" using computer software. 4) Customized microstereotactic frames were rapid-prototyped to serve as drill guides constraining the drill to follow the appropriate path. 5) During cochlear implant surgery, after drilling of the facial recess, drill guides were mounted on the bone-implanted anchors. 6) Accuracy of paths was assessed via intraoperative photodocumentation. RESULTS: All surgical paths successfully traversed the facial recess and hit the basal turn of the cochlea. Distance in millimeters (average +/ SD) from the midpoint of the drill to the facial nerve was 1.18 +/- 0.68 for the "manual" path and 1.24 +/- 0.44 mm for the "automatic" path and for the chorda tympani 0.986 +/- 0.48 for the "manual" path and 1.22 +/- 0.62 for the "automatic" path. CONCLUSIONS: Percutaneous cochlear implant access using customized drill guides based on preoperative CT scans and image-guided surgery technology can be safely accomplished. PMID- 18401280 TI - Soluble intercellular adhesion molecule 1 and soluble vascular cell adhesion molecule 1 in sudden hearing loss. AB - HYPOTHESIS: The aim of the present study was to evaluate the concentration of soluble intercellular adhesion molecule 1 and soluble vascular cell adhesion molecule 1 in patients affected by sudden sensorineural hearing loss (SSHL). STUDY DESIGN: Prospective study. SETTING: Tertiary referral center. PATIENTS: Patients affected by SSHL were evaluated. Inclusion criteria for this study were hearing loss of more than 30 dB hearing level affecting at least 3 contiguous frequencies, normal hearing on the contralateral ear, negative history of hearing loss or ear surgery in the affected ear, and magnetic resonance with gadolinium negative for VIII cranial nerve pathologic findings. INTERVENTION: Circulating levels of soluble intercellular adhesion molecule 1 and soluble vascular cell adhesion molecule (VCAM) 1 were evaluated by means of enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: The levels of adhesion molecules in SSHL patients were compared with those of a control group. RESULTS: Intercellular adhesion molecule 1 and VCAM-1 levels in sera of patients with SSHL were significantly higher than those of the matched control subjects (p < 0.001). Statistical analysis did not show significant differences between the 2 groups in terms of the known vascular risk factors such as total and fractionated cholesterol, triglycerides, fibrinogen, erythrocyte sedimentation rate smoking, and diabetes. CONCLUSION: The results of this study show that in SSHL patients, there is an increased expression of circulating adhesion molecules confirming the existence of an endothelial dysfunction and supporting the vascular involvement in the pathogenesis of the disease. The identification of high levels of adhesion molecules and of the endothelial dysfunction open the way to selective pharmacologic treatments able to correct the activation of endothelial cells. PMID- 18401281 TI - Surgical factors in pediatric cochlear implantation and their early effects on electrode activation and functional outcomes. AB - OBJECTIVE: To assess the impact of surgical factors on electrode status and early communication outcomes in young children in the first 2 years of cochlear implantation. STUDY DESIGN: Prospective multicenter cohort study. SETTING: Six tertiary referral centers. PATIENTS: Children 5 years or younger before implantation with normal nonverbal intelligence. INTERVENTION: Cochlear implant operations in 209 ears of 188 children. MAIN OUTCOME MEASURES: Percent active channels, auditory behavior as measured by the Infant Toddler Meaningful Auditory Integration Scale/Meaningful Auditory Integration Scale and Reynell receptive language scores. RESULTS: Stable insertion of the full electrode array was accomplished in 96.2% of ears. At least 75% of electrode channels were active in 88% of ears. Electrode deactivation had a significant negative effect on Infant Toddler Meaningful Auditory Integration Scale/Meaningful Auditory Integration Scale scores at 24 months but no effect on receptive language scores. Significantly fewer active electrodes were associated with a history of meningitis. Surgical complications requiring additional hospitalization and/or revision surgery occurred in 6.7% of patients but had no measurable effect on the development of auditory behavior within the first 2 years. Negative, although insignificant, associations were observed between the need for perioperative revision of the device and 1) the percent of active electrodes and 2) the receptive language level at 2-year follow-up. CONCLUSION: Activation of the entire electrode array is associated with better early auditory outcomes. Decrements in the number of active electrodes and lower gains of receptive language after manipulation of the newly implanted device were not statistically significant but may be clinically relevant, underscoring the importance of surgical technique and the effective placement of the electrode array. PMID- 18401282 TI - Cochlear otosclerosis and endolymphatic hydrops. PMID- 18401283 TI - Is there a right cochlear implant advantage? AB - OBJECTIVE: To compare behavioral speech perception performance of children with right versus left cochlear implants (CIs). STUDY DESIGN: A retrospective cohort study. SETTING: Academic university medical center. SUBJECTS: Seventy-one prelingually deafened children that obtained a CI device at 48 months or younger. INTERVENTION: Cochlear implantation with Cochlear, Advanced Bionics, and Med-El devices. MAIN OUTCOME MEASUREMENTS: Patients were divided into 2 groups according to ear of implant (right, n = 30; left, n = 41) and matched in age at implantation and preoperative audiologic results. Multivariate analysis of variance was used to evaluate the effects of 1) side of CI, 2) age at implantation, 3) time with CI (T1, 18-24 mo; T2, 36-42 mo), and 4) dominance (i.e., compatibility between CI side and handedness) on performance in a monosyllabic open-set test scored for words and phonemes. RESULTS: A small yet significant "right CI advantage" was evident throughout the study follow-up and was independent of age at implantation. The performance of children implanted at 24 months or younger was significantly higher than that of children implanted between 25 and 48 months. Regardless of CI side and age at implantation, all children exhibited improvement in speech perception with continuous use. CONCLUSION: The present study provides first-time evidence for a right CI advantage for speech perception in prelingually deafened children that can be taken into account when selecting side of CI in candidates with similar residual hearing in both ears and no anatomic constraints. The present data lend further support to the notions that greater gains in speech perception are associated with earlier age at implantation and continuous use. PMID- 18401284 TI - Ear pain and its treatment in hypobaric chamber training in the Japan Air Self Defense Force. AB - BACKGROUND: We have documented that ear pain is the most prevalent physiologic incident during hypobaric chamber training in the Japan Air Self-Defense Force. Ear pain may increase also in flight in the future because it is closely related to allergic rhinitis. Therefore, it is very important to know the characteristic of ear pain and the efficacy of its treatment. METHODS: The incidence of ear pain was calculated in each training profile from 1990 to 1998. Type III chamber flight records were further analyzed for the characteristics of ear pain: relationship with a trainee occupational category, time of occurrence of ear pain, and efficacy of treatment. RESULTS: Of 17,935 exposures, 740 trainees (4.1%) had ear pain. Of 7,047 trainees, 429 (6.1%) complained of ear pain especially in Type III, totaling 625 times. Fighter pilots and cargo pilots complained of ear pain one twelfth and one third the number of times, respectively, compared with passengers. Of the 625 episodes, 616 (98.6%) occurred during descent. Three kinds of treatment were administered until the pain was relieved in the following order: Valsalva maneuver, Politzer bag, and decompression. The efficacy rates were 35.8, 92.3, and 83.9%, respectively. Only 5 trainees (0.07%) could not complete training due to ear pain despite treatment. CONCLUSION: The combination treatment of Valsalva maneuver, Politzer bag, and decompression is very effective for relieving ear pain encountered during hypobaric chamber training. A health specialist needs to understand ear pain and its treatment in hypobaric environment such as aircraft. PMID- 18401285 TI - Combination therapy (intratympanic dexamethasone + high-dose prednisone taper) for the treatment of idiopathic sudden sensorineural hearing loss. AB - BACKGROUND: Idiopathic Sudden Sensorineural Hearing Loss (ISSNHL), commonly defined as greater than 20 dB of unilateral hearing loss in at least 3 frequencies occurring within 3 days, has a reported incidence of 5 to 20 per 100,000 patients per year. Untreated, it has a recovery rate of 32 to 65%. Although accepted therapy is high-dose prednisone taper (HDPT), recent publications suggest that intratympanic dexamethasone (IT-Dex) therapy may improve hearing recovery. METHODS: This multicenter, double-blinded, placebo controlled, randomized study seeks to compare hearing results in ISSNHL patients who have received HDPT alone, IT-Dex alone, or IT-Dex and HDPT (combination therapy). Fifty-one patients with a less than 6-week history of ISSNHL were randomized to 1 of 3 arms and followed prospectively. Group A (17 patients) received IT-Dex therapy with placebo taper, whereas Group B (18 patients) were administered HDPT and placebo intratympanic injections. Patients in Group C (16 patients) were administered IT-Dex and HDPT, otherwise known as combination therapy. Injections (IT-Dex/placebo) and audiograms were performed weekly for 3 weeks, and a final audiogram was obtained 4 weeks after the final injection. RESULTS: Patients receiving combination therapy (IT-Dex + HDPT) in Group C had an average improvement in speech discrimination score of 44 percentage points and a 40-dB improvement in pure-tone average (PTA). Patients in Group C had statistically significant improvements in speech discrimination score compared with Group B patients (HDPT alone; p < 0.05). When defining a significant improvement in PTA as greater than 15 dB, there was a statistically significant difference between the groups in the proportion of patients achieving hearing improvement. Furthermore, the proportion of patients achieving a significant PTA improvement in Group C was statistically greater than patients in Group B (p < 0.02). Logistic regression analysis indicates that patients receiving combination therapy demonstrated better odds of hearing recovery than patients in both of the other groups (p < 0.05), when all 3 groups were adjusted for age, vertigo, initial hearing levels, and time delay between onset of hearing loss and treatment. Lastly, combination therapy patients recovered their hearing more quickly than patients in the other groups (p < 0.05). CONCLUSION: The results of this study suggest that ISSNHL patients treated with IT-Dex + HDPT (combination therapy) have a higher likelihood of hearing recovery than those treated with HDPT alone. PMID- 18401286 TI - Intralabyrinthine hemorrhage associated with superficial siderosis of the central nervous system. PMID- 18401287 TI - The impact of cochlear implants from the perspective of significant others of adult cochlear implant users. AB - OBJECTIVE: To explore the benefits and shortcomings of cochlear implants as perceived by a cochlear implant user's significant other in the context of the World Health Organization's International Classification of Functioning, Disability and Health (ICF). STUDY DESIGN: An open-ended questionnaire concerning the perceived shortcomings and benefits of the implant was completed by the significant others of cochlear implants users. SETTING: The study involved the participants of an adult cochlear implant program. SUBJECTS: Significant others as selected by the cochlear implant user. MAIN OUTCOME MEASURE: Nature of effects reported by the significant other relating to both the cochlear implant user and the significant other using the ICF domains. RESULTS: Benefits experienced by the significant others included a decrease in communication burden and an increased participation in life activities. Those affecting the cochlear implant user were related to psychological issues, particularly an increase in confidence. Improved quality of life was reported for both partners. Reported shortcomings predominantly involved the cochlear implant user and were device related. CONCLUSION: Because family and friends play an important role in how those with a profound hearing impairment cope with the impairment, it is important to acknowledge the impact that an intervention has on those supportive individuals. This study shows that a cochlear implant provides psychosocial and acoustic benefits for the significant other and the cochlear implant user. The ICF was able to accommodate both the auditory and nonauditory impact of the cochlear implant and the residual hearing difficulty. PMID- 18401288 TI - DMSA renal scans and the top-down approach to urinary tract infection. AB - Assessment of first febrile urinary tract infection in young children has usually involved 2 imaging techniques: renal ultrasonogram and voiding cystourethrography. Currently, there is growing interest in using the dimercaptosuccinic acid (DMSA) scan labeled with technetium-99m as an alternative initial study, in the evaluation of UTI. DMSA renal scanning is the most sensitive radiologic study to detect acute pyelonephritis. Early DMSA renal scanning has been called the top-down approach, because the focus is the identification of kidney injury rather than reflux. Positive and negative aspects of DMSA renal scans are discussed by a pediatric urologist. PMID- 18401289 TI - Safety of the intranasal, trivalent, live attenuated influenza vaccine (LAIV) in children with intermittent wheezing in an open-label field trial. AB - BACKGROUND: Safety of the intranasal, trivalent, live attenuated influenza vaccine (LAIV) in children with asthma is unknown. A previous report showed an "asthma signal" in children aged 18-35 months. METHODS: Healthy children aged 1.5 18 years with history of intermittent wheezing received single annual LAIV doses during a 4-year trial. Rates of medically-attended acute respiratory illnesses, including acute asthma exacerbation, during 0-14 and 0-42 days post-LAIV were compared with respective reference periods (before day 0 and after 14 or 42 days). To assess the risk of new-onset asthma, LAIV recipients without history of wheezing were analyzed. RESULTS: During each of the 4 years, 454, 656, 656, and 430 children, respectively, with intermittent wheezing who received LAIV had no increased risk for medically-attended acute respiratory illnesses, including asthma exacerbation. First-dose LAIV recipients, including those aged 1.5-4 years, and those receiving 2-4 consecutive annual doses had no increased risk. Children with parents' report of intermittent wheezing and those with administrative database codes for asthma during 2 prior years had no increased risk. During the 4 years, 2952, 3092, 2953, and 2478 children without history of wheezing had no increased risk of new-onset asthma. CONCLUSIONS: LAIV administration in children aged 1.5-18 years with history of intermittent wheezing was safe, and was not associated with increased risk for medically attended acute respiratory illnesses, including acute asthma exacerbation. This was true for the first and 2-4 consecutive annual doses. Parents' report of intermittent wheezing was reliable. First-dose LAIV was not associated with new onset asthma in children without history of wheezing. PMID- 18401290 TI - [Drusen in children: three case studies]. AB - We report three cases of optic disc drusen in children. The children were 5 months, 10 years, and 11 years old. Two of them were male. The optic disc drusen was unilateral in two cases and bilateral in one case. One child had a history of Down syndrome and another one a history of Alagille syndrome. The third child had no particular general history but he had unilateral high myopia with optic disc drusen in the same eye. Fundus examination showed pseudopapilledema in all three cases; the disc appeared elevated and its margins were blurred. Ultrasonography of the eye demonstrated optic disc drusen in the three cases. Optic disc calcification on CT scan was identified in only one case. In children, optic disc drusen are often buried, leading to pseudopapilledema appearance of the optic disc. Ultrasonography is highly sensitive in detecting optic disc drusen, making more expensive investigations unnecessary. PMID- 18401291 TI - [Retinal neovascularization with myelinated nerve fibers]. AB - INTRODUCTION: The prevalence of myelinated retinal nerve fibers is 0.3%-0.6% of eyes. Although they often constitute benign lesions, they are rarely associated with retinal vascular abnormalities including preretinal neovascularization. CASE REPORT: A 31-year-old patient, with no previous pathological antecedents, consulted us for myodesopsia in her left eye. The exam found a visual acuity of 10/10 and a normal anterior segment in the two eyes. At the ophthalmoscopic examination of the left eye, we noted myelinated nerve fibers in the inferotemporal quadrant that were associated with a temporal neovascular bouquet covering area of 1.5 optic disks. A small amount of intravitreal hemorrhage was found. The ophthalmoscopic examination of the right eye was normal. The treatment consisted in sector-based photocoagulation with argon laser and cryotherapy. DISCUSSION: The source of neovascularization in the myelinated retinal nerve fibers was discussed. Underlying retinal ischemia is the most probable mechanism. The treatment was based on sector-based or panretinal photocoagulation. CONCLUSION: Preretinal neovascularization is a rare complication of myelinated nerve fibers. The earlier the care is given, the more complications can be avoided. PMID- 18401292 TI - [Familial exudative vitreoretinopathy associated with persistence of hyaloid artery]. AB - Familial exudative vitreoretinopathy (FEVR) is a well-defined inherited disorder of retinal vessel development. Association of FEVR with other congenital ocular diseases has already been described. We report the first clinical case of FEVR associated with hyaloid artery persistence. An 18-year-old Tunisian woman and four members of her family (mother, two brothers, and sister) presented signs of FEVR. Examination of this patient also revealed functional hyaloid artery persistence in her right eye. These two disorders have many common aspects in their pathological process, in particular vascular endothelial growth factor expression, which can explain this association. PMID- 18401293 TI - [Pseudoexfoliation syndrome in cataract surgery. Retrospective study of 37 cases]. AB - PURPOSE: To analyze the clinical features of pseudoexfoliation syndrome and to determine complications in cataract surgery in eyes with pseudoexfoliation syndrome compared with eyes without this syndrome. METHODS: A retrospective, single-center comparative study was conducted on patients who underwent cataract surgery between 2004 and 2006. Demographic parameters (age, sex, ethnicity, medical and ophthalmologic history), clinical features (visual acuity, pupil dilatation, intraocular pressure) before and after surgery, and surgical complications were analyzed and compared to a control group without pseudoexfoliation. RESULTS: Seventy-four eyes of 71 patients were included in the study. The sex ratio of the pseudoexfoliation group was 1.26 with 19 males (56%) and 15 females (44%). Pseudoexfoliation was unilateral in 21 cases (61%). Open angle glaucoma was associated in 11 cases (32%) in the pseudoexfoliation group versus two cases in the control group (5.4%). Poor pupil dilatation was observed in eight cases (24%) in the pseudoexfoliation group and in one case (2.7%) in the control group. Phacoemulsification with IOL was the surgical technique in all the cases with only one case (2.7%) of anterior chamber IOL in the pseudoexfoliation group. There was no significant difference in the surgical complications between the pseudoexfoliation and control group (5.4%). The mean preoperative intraocular pressure was 16+/-5 mmHg in the pseudoexfoliation group and 14+/-3 mmHg in the control group. The mean postoperative intraocular pressure at 1 month was 14+/-4 mmHg in the pseudoexfoliation group and 14+/-3 in the control group. The mean preoperative visual acuity was 1.0+/-0.7 LogMar in the pseudoexfoliation group and 0.7+/-0.5 LogMar in the control group. The mean postoperative visual acuity was 0.4+/-0.6 LogMar in the pseudoexfoliation group and 0.2+/-0.1 LogMar in the control group. The mean follow-up was 70 days in the pseudoexfoliation group and 30 days in the control group. CONCLUSION: Pseudoexfoliation syndrome is the most common in its unilateral presentation. It is frequently associated with chronic secondary open-angle glaucoma. Poor pupil dilatation is one of the most common problems faced by cataract surgeons. Nevertheless, pseudoexfoliation syndrome did not confer a statistically higher risk for surgical complication in eyes without marked phacodonesis or lens subluxation. PMID- 18401294 TI - [Trabeculectomy according to P. Khaw's protocol: medium-term tonometric results]. AB - PURPOSE: P. Khaw has described a new trabeculectomy technique to enhance the reliability (fewer complications) and the efficacy of the classic trabeculectomy introduced by Cairns. The purpose of this study was to assess the medium-term tonometric results and complications of this new surgical procedure. MATERIAL: and methods: In a retrospective and nonrandomized study, 26 consecutive eyes of 21 patients who underwent trabeculectomy according to P. Khaw's protocol were analyzed. The surgeon conducted postoperative care for the first 2 months. The minimal follow-up was 1 year, with a mean follow-up of 16.8 months (range, 12-18 months). The criteria for success were defined as intraocular pressure (IOP) less than 16 mmHg without treatment. RESULTS: The mean IOP remained stable around 12 mmHg during the study follow-up: 12.5 mmHg+/-3.07 at 12 months and 11.8 mmHg+/ 3.17 at 18 months. The success rate was 92.3% at 3 months and 84.6% at 6, 9, and 12 months. The mean IOP reduction was 44.1%+/-17.05. No failure was observed between months 6 and 18. The probability for an IOP under 16 mmHg without treatment at months 12 and 18 was 84% (Kaplan-Meier). At 1 year, 65% of the eyes had an IOP under 14 mmHg without treatment. Complications were limited and transient: one case of choroidal detachment and three cases of hyphema. No flat anterior chamber and no infection were observed during follow-up. CONCLUSION: The new trabeculectomy described by P. Khaw is a promising procedure because at the medium term it provides tonometric results matching the target pressure of glaucomatous patients without the complications of the classic trabeculectomy. PMID- 18401295 TI - [Evaluation of the results of 12-mm recession of the inferior oblique muscle in superior oblique palsy: retrospective study in six patients]. AB - INTRODUCTION: Surgical possibilities in superior oblique palsy are numerous. The aim of this study was to evaluate the results of a 12-mm inferior oblique recession in each cardinal position of gaze. PATIENTS AND METHODS: Six patients suffering from superior oblique palsy were studied. All of them presented disabling symptoms such as right-left incomitance, and hypertropia in primary gaze equal to or lower than 25 diopters. They all underwent a pre- and postoperative deviometry. The surgical protocol was a 12-mm inferior oblique muscle recession, with reinsertion 4 mm behind the inferior rectus. RESULTS: Functional results were positive. Mean hypertropia in primary gaze was 14.7 diopters for far and 12.7 diopters for near vision. The reduction of hypertropia in primary gaze was approximately 10 diopters, corresponding to 71.6% for far and 81.4% hypertropia for near vision. The mean postoperative vertical deviation was equal to 4.2 diopters for far and 2.3 diopters for near vision. Results in adduction were good even if there was a high level of hypertropia in this gaze position. CONCLUSION: Many authors suggest considering anatomic abnormalities to improve surgical results such as traction testing on the oblique superior and superior rectus muscles. We can suggest a first-intention 12-mm recession of the inferior oblique muscle in cases of right-left incomitance with hypertropia in primary gaze lower than 25 diopters. PMID- 18401296 TI - [Outcome of photodynamic therapy for subretinal neovascularization secondary to choroidal nevi]. AB - INTRODUCTION: Choroidal nevi are the most common benign intraocular tumors. However, the occurrence of subretinal neovascularization secondary to choroidal nevus is very rare. Our goal was to assess the efficacy and the safety of photodynamic therapy (PDT) for treatment of subretinal neovascularization related to choroidal nevi. PATIENTS AND METHOD: We conducted a retrospective study including subretinal neovascularization secondary to choroidal nevi treated by PDT from 1999 to 2005. PDT was performed according to the standard protocols used for treating neovascularization due to age-related macular degeneration (ARMD). Post-PDT follow-up was also done according to the guidelines used in ARMD. Moreover, tumors were followed up every 3 months. RESULTS: Six subretinal neovascularizations secondary to choroidal nevi were included in our study. All nevi were localized in the posterior choroids. The median diameter of tumors was 3mm and their thickness was less than 2mm in all cases. Subretinal neovascularizations were classic in five cases. The last case was a small vascularized pigment epithelium detachment. Locations were subfoveal in two cases, juxtafoveal in three cases, and extrafoveal in one case. In half the cases, the size of subretinal neovascularization was 1-disk-diameter or less. A mean of 3.3 sessions of PDT were necessary to obtain the stabilization of subretinal neovascularization on both fluorescein angiography and OCT. The average follow-up period was 35 months with no patients lost to follow-up. Visual outcomes were extremely variable. Large or subfoveal neovascularization but also a history of atrophic ARMD were related to worse functional results. No growth of the nevi was observed. CONCLUSIONS: Subretinal neovascularization is reassuring from the oncologist's point of view because it is considered a relative indicator of benignity of the nevus. However, it often marks a definitive turning point in visual function. PDT appears to be a safe and effective procedure for the treatment subretinal neovascularization secondary to choroidal nevus, with results similar to those observed in ARMD. PMID- 18401297 TI - Retinal detachment: visual acuity and subretinal immunoreactive endothelin-1. AB - PURPOSE: To analyze whether subretinal (SRF) endothelin-1 (ET-1) - a vasoactive, mitogenic, and pro-apoptotic peptide - levels are related to visual acuity (VA) in rhegmatogenous retinal detachment (RD). PATIENTS AND METHODS: Sixty-six healthy patients between 42 and 70 years of age with unilateral RD, all candidates for scleral buckling surgery (PVRT, c.602C>T, and c.808G>T) showed significant differences in metformin pharmacokinetics when compared with the reference genotype, with higher peak plasma concentration (C(max)) and area under the curve (AUC) and lower renal clearance (Cl(renal)), thereby suggesting that a decrease in transport function associated with the SLC22A2 variants results in reduced Cl(renal) of metformin and consequently leads to increased plasma concentrations. PMID- 18401340 TI - Clinical pharmacology: opportunities for educating new practitioners--an Australian perspective. AB - There has been a transformation in the role of clinical pharmacologists in Australasia. The traditional approach was for medically qualified, college accredited (postgraduate education), hospital-based clinical pharmacologists to confine themselves mainly to a local focus. Today many more opportunities exist for expanding the roles of health professionals who have the clinical pharmacology training. These professionals can influence national and international policy, practice, and education in their field. However, the new roles do require extra initiatives in providing educational input in clinical pharmacology to take care of future needs. Some of the potential gaps in the process are identified in this article. PMID- 18401341 TI - Upper intestinal lipids trigger a gut-brain-liver axis to regulate glucose production. AB - Energy and glucose homeostasis are regulated by food intake and liver glucose production, respectively. The upper intestine has a critical role in nutrient digestion and absorption. However, studies indicate that upper intestinal lipids inhibit food intake as well in rodents and humans by the activation of an intestine-brain axis. In parallel, a brain-liver axis has recently been proposed to detect blood lipids to inhibit glucose production in rodents. Thus, we tested the hypothesis that upper intestinal lipids activate an intestine-brain-liver neural axis to regulate glucose homeostasis. Here we demonstrate that direct administration of lipids into the upper intestine increased upper intestinal long chain fatty acyl-coenzyme A (LCFA-CoA) levels and suppressed glucose production. Co-infusion of the acyl-CoA synthase inhibitor triacsin C or the anaesthetic tetracaine with duodenal lipids abolished the inhibition of glucose production, indicating that upper intestinal LCFA-CoAs regulate glucose production in the preabsorptive state. Subdiaphragmatic vagotomy or gut vagal deafferentation interrupts the neural connection between the gut and the brain, and blocks the ability of upper intestinal lipids to inhibit glucose production. Direct administration of the N-methyl-d-aspartate ion channel blocker MK-801 into the fourth ventricle or the nucleus of the solitary tract where gut sensory fibres terminate abolished the upper-intestinal-lipid-induced inhibition of glucose production. Finally, hepatic vagotomy negated the inhibitory effects of upper intestinal lipids on glucose production. These findings indicate that upper intestinal lipids activate an intestine-brain-liver neural axis to inhibit glucose production, and thereby reveal a previously unappreciated pathway that regulates glucose homeostasis. PMID- 18401342 TI - NF-kappaB dictates the degradation pathway of IkappaBalpha. AB - IkappaB proteins are known as the regulators of NF-kappaB activity. They bind tightly to NF-kappaB dimers, until stimulus-responsive N-terminal phosphorylation by IKK triggers their ubiquitination and proteasomal degradation. It is known that IkappaBalpha is an unstable protein whose rapid degradation is slowed upon binding to NF-kappaB, but it is not known what dynamic mechanisms control the steady-state level of total IkappaBalpha. Here, we show clearly that two degradation pathways control the level of IkappaBalpha. Free IkappaBalpha degradation is not controlled by IKK or ubiquitination but intrinsically, by the C-terminal sequence known as the PEST domain. NF-kappaB binding to IkappaBalpha masks the PEST domain from proteasomal recognition, precluding ubiquitin independent degradation; bound IkappaBalpha then requires IKK phosphorylation and ubiquitination for slow basal degradation. We show the biological requirement for the fast degradation of the free IkappaBalpha protein; alteration of free IkappaBalpha degradation dampens NF-kappaB activation. In addition, we find that both free and bound IkappaBalpha are similar substrates for IKK, and the preferential phosphorylation of NF-kappaB-bound IkappaBalpha is due to stabilization of IkappaBalpha by NF-kappaB. PMID- 18401343 TI - Quantitative proliferation dynamics and random chromosome segregation of hair follicle stem cells. AB - Regulation of stem cell (SC) proliferation is central to tissue homoeostasis, injury repair, and cancer development. Accumulation of replication errors in SCs is limited by either infrequent division and/or by chromosome sorting to retain preferentially the oldest 'immortal' DNA strand. The frequency of SC divisions and the chromosome-sorting phenomenon are difficult to examine accurately with existing methods. To address this question, we developed a strategy to count divisions of hair follicle (HF) SCs over time, and provide the first quantitative proliferation history of a tissue SC during its normal homoeostasis. We uncovered an unexpectedly high cellular turnover in the SC compartment in one round of activation. Our study provides quantitative data in support of the long-standing infrequent SC division model, and shows that HF SCs do not retain the older DNA strands or sort their chromosome. This new ability to count divisions in vivo has relevance for obtaining basic knowledge of tissue kinetics. PMID- 18401344 TI - A molecular switch required for retrovirus assembly participates in the hexagonal immature lattice. AB - In the Rous sarcoma virus (RSV) Gag protein, the 25 amino-acid residues of the p10 domain immediately upstream of the CA domain are essential for immature particle formation. We performed systematic mutagenesis on this region and found excellent correlation between the amino-acid side chains required for in vitro assembly and those that participate in the p10-CA dimer interface in a previously described crystal structure. We introduced exogenous cysteine residues that were predicted to form disulphide bonds across the dimer interface. Upon oxidation of immature particles, a disulphide-linked Gag hexamer was formed, implying that p10 participates in and stabilizes the immature Gag hexamer. This is the first example of a critical interaction between two different Gag domains. Molecular modeling of the RSV immature hexamer indicates that the N-terminal domains of CA must expand relative to the murine leukaemia virus mature hexamer to accommodate the p10 contact; this expansion is strikingly similar to recent cryotomography results for immature human immunodeficiency virus particles. PMID- 18401345 TI - Metaplasticity: tuning synapses and networks for plasticity. AB - Synaptic plasticity is a key component of the learning machinery in the brain. It is vital that such plasticity be tightly regulated so that it occurs to the proper extent at the proper time. Activity-dependent mechanisms that have been collectively termed metaplasticity have evolved to help implement these essential computational constraints. Various intercellular signalling molecules can trigger lasting changes in the ability of synapses to express plasticity; their mechanisms of action are reviewed here, along with a consideration of how metaplasticity might affect learning and clinical conditions. PMID- 18401347 TI - How to succeed in science: a concise guide for young biomedical scientists. Part II: making discoveries. AB - Making discoveries is the most important part of being a scientist, and also the most fun. Young scientists need to develop the experimental and mental skill sets that enable them to make discoveries, including how to recognize and exploit serendipity when it strikes. Here, I provide practical advice to young scientists on choosing a research topic, designing, performing and interpreting experiments and, last but not least, on maintaining your sanity in the process. PMID- 18401348 TI - How to succeed in science: a concise guide for young biomedical scientists. Part I: taking the plunge. AB - Biomedical research has never been more intellectually exciting or practically important to society. Ironically, pursuing a career as a biomedical scientist has never been more difficult. Here I provide unvarnished advice for young biomedical scientists on the difficulties that lie ahead and on how to find the right laboratories for training in the skills that you will need to succeed. Although my advice is geared towards succeeding in the United States, many aspects apply to other countries. PMID- 18401346 TI - Adipocyte dysfunctions linking obesity to insulin resistance and type 2 diabetes. AB - Acquired resistance to the action of insulin to stimulate glucose transport in skeletal muscle is associated with obesity and promotes the development of type 2 diabetes. In skeletal muscle, insulin resistance can result from high levels of circulating fatty acids that disrupt insulin signalling pathways. However, the severity of insulin resistance varies greatly among obese people. Here we postulate that this variability might reflect differences in levels of lipid droplet proteins that promote the sequestration of fatty acids within adipocytes in the form of triglycerides, thereby lowering exposure of skeletal muscle to the inhibitory effects of fatty acids. PMID- 18401349 TI - Obesity and sexual dysfunction, male and female. AB - Obesity has become a worldwide public health problem of epidemic proportions, as it may decrease life expectancy by 7 years at the age of 40 years: excess bodyweight is now the sixth most important risk factor contributing to the overall burden of disease worldwide. Overweight and obesity may increase the risk of erectile dysfunction (ED) by 30-90% as compared with normal weight subjects. On the other hand, subjects with ED tend to be heavier and with a greater waist than subjects without ED, and also are more likely to be hypertensive and hypercholesterolemic. The metabolic syndrome, characterized by a clustering of risk factors associated with insulin resistance and abdominal obesity, associates with ED. Moreover, women with the metabolic syndrome have an increased prevalence of sexual dysfunctions as compared with matched control women. Lifestyle changes aimed at reducing body weight and increasing physical activity induce amelioration of both erectile and endothelial functions in obese men. Moreover, preliminary evidence suggests that a Mediterranean-style diet might be effective in ameliorating sexual function in women with the metabolic syndrome. Lifestyle changes, mainly focussing on regular physical activity and a healthy diet, are effective and safe ways to reduce cardiovascular diseases and premature mortality in all population groups; they may also prevent and treat sexual dysfunctions in both sexes. PMID- 18401350 TI - Prophylactic antibiotics in the prevention of catheter-associated bloodstream bacterial infection in preterm neonates: a systematic review. AB - OBJECTIVE: To evaluate the efficacy of prophylactic antibiotics in preventing infection associated with central venous catheters in preterm neonates. STUDY DESIGN: The search strategy of the Cochrane Neonatal Review Group was used. The following databases were searched: Medline, Cochrane Central Register of Controlled Trials, CINAHL and EMBASE. In addition, we hand-searched abstracts of Pediatric Academic Societies annual meetings published in Pediatric Research (1990 to July 2007) and Canadian Pediatric Society annual meeting proceedings (1990 to July 2007). No language restrictions were applied. Included were randomized controlled trials of antibiotics given prophylactically to prevent infection in preterm infants (<37 completed weeks) less than 1-month old admitted to neonatal intensive care units. Both centrally or peripherally inserted central venous catheters were included. Assessment of methodological quality and extraction of data for included trials was undertaken independently by two authors. When suitable, data from trials were combined in a meta-analysis. RESULT: A total of three studies were found which addressed the role of prophylactic antibiotics to prevent catheter-related infection in neonates. Two studies used vancomycin as the prophylactic antibiotic and one study used amoxicillin. The meta-analysis of studies that used vancomycin had shown an absolute risk reduction of infection from 23 to 2.4%, which yields a number needed to treat equal to 5 (P=0.0001). Total duration of catheter stay and mortality, were both similar in the vancomycin and control groups. In the amoxicillin study, catheter-related sepsis was not significantly different between the treatment and control groups (P=0.40). The rate of colonization, however, was significantly higher in the control group (relative risk 0.48; 95% CI 0.12, 1.35). The incidence of necrotizing enterocolitis, intracranial hemorrhage, thrombosis and deaths were not statistically significant between groups. CONCLUSION: Prophylactic vancomycin appeared to be effective in preventing catheter-related sepsis in preterm neonates. The potential risks, however, of the emergence of resistance because of prophylactic antibiotics, and their continued effectiveness, need further evaluation, before routine use can be recommended. PMID- 18401351 TI - Amerindian ancestry in Argentina is associated with increased risk for systemic lupus erythematosus. AB - Previous studies have demonstrated that in admixed populations, West African ancestry is associated with an increased prevalence of systemic lupus erythematosus (SLE). In the current study, the effect of Amerindian ancestry in SLE was examined in an admixed population in Argentina. The Argentine population is predominantly European with approximately 20% Amerindian admixture, and a very small (<2%) contribution from West Africa. The results indicate that Amerindian admixture in this population is associated with a substantial increase in SLE susceptibility risk (Odds Ratio=7.94, P=0.00006). This difference was not due to known demographic factors, including site of collection, age and gender. In addition, there were trends towards significance for Amerindian ancestry influencing renal disease, age of onset and anti-SSA antibodies. These studies suggest that populations with Amerindian admixture, like those with West African admixture, should be considered in future studies to identify additional allelic variants that predispose to SLE. PMID- 18401352 TI - EVI5 is a risk gene for multiple sclerosis. AB - HLA-DRB1 is the major locus associated with risk for multiple sclerosis (MS). A recent genome-wide study showed three additional single-nucleotide polymorphisms (SNPs), within the IL2RA and IL7RA genes respectively, also to be associated with MS. Consistent association but lower significance was found for 13 other SNPs. In this study, we aimed to verify association of these SNPs with MS in 46 MS patients and 194 controls from a Dutch genetically isolated population. Apart from the human leukocyte antigen locus, the EVI5 gene on chromosome 1 was confirmed as a novel risk gene, with odds ratios (ORs) even higher than those from the MS Consortium (ORs 2.01 and 1.9; P=0.01). The risk effect of EVI5 was further validated for the general MS population in an independent set of 1318 MS patients from the Canadian Collaborative Project on the Genetic Susceptibility to MS. On the basis of the transmission disequilibrium testing, a weak but significant risk effect was observed (OR 1.15; P=0.03 and OR 1.15; P=0.04). This study confirms EVI5 as another risk locus for MS; however, much of the genetic basis of MS remains unidentified. PMID- 18401354 TI - Analysis of the functional relevance of a putative regulatory SNP of PDCD1, PD1.3, associated with systemic lupus erythematosus. AB - This study aimed to test the functional effects of the PD1.3 single nucleotide polymorphism (SNP) (rs11568821), which were proposed based on its association to systemic lupus erythematosus (SLE) susceptibility and in electrophoretic mobility shift assays (EMSA) results. We analysed transcriptional effects of the PD1.3 locus by enhancer reporter assays. Results were against the hypothesis that the PD1.3 locus acts as enhancer in transcriptional regulation of PDCD1. In addition, they excluded a differential effect of the PD1.3 alleles. EMSA results confirmed that oligonucleotides with the PD1.3 G allele bind RUNX1 but not those with the A allele. However, binding to PD1.3 G oligonucleotides was much lower than binding to positive control oligonucleotides. Criss-cross experiments showed that this was due to flanking nucleotides in the PD1.3 sequence that negatively affect RUNX1 binding. These results cast doubts on the functional relevance of the PD1.3 SNP and, together with the lack of association in several studies, put into question its role as an SLE susceptibility factor. Investigation of other PDCD1 polymorphisms is needed to uncover the possible effect of this gene on SLE susceptibility. PMID- 18401358 TI - Defining 'natural'. PMID- 18401353 TI - Stat4-dependent, T-bet-independent regulation of IL-10 in NK cells. AB - Interleukin-10 (IL-10) is intensely studied, yet little is known about the mechanisms that control IL-10 expression. We identified striking similarities between IL-10 and interferon-gamma (IFN-gamma) regulation in mouse natural killer (NK) cells. Like IFN-gamma, IL-10 expression is induced by IL-2 and IL-12 and IL 2+IL-12 stimulation is synergistic. Unlike IFN-gamma, neither IL-18 nor Ly-49D cross-linking induced IL-10 expression however. Additionally, the IL-12 homologs IL-23 and IL-27 also do not regulate NK cell-specific IL-10. We determined that a small population of NK cells accounts for IL-10 production. The induction of IL 10 by IL-2+IL-12 treatment in NK cells appears to be biphasic, with an initial burst of expression which diminishes by 12 h but spikes again at 18 h. We determined that much like IFN-gamma, Stat4 is largely required for IL-12-induced IL-10. Conversely, we observed normal induction of IL-10 in T-bet-deficient NK cells. We identified a Stat4-binding element in the fourth intron of the Il10 gene, which is completely conserved between mouse and human. This intronic Stat4 motif is within a conserved noncoding sequence, which is also a target for cytokine-induced histone acetylation. These findings highlight tissue- and receptor-specific IL-10 regulatory mechanisms, which may be part of an early feedback loop. PMID- 18401359 TI - The path to productive partnerships. PMID- 18401360 TI - Ready or not. PMID- 18401363 TI - Advanced biofuels face an uncertain future. PMID- 18401364 TI - FDA to vet embryonic stem cells' safety. PMID- 18401366 TI - The fraudster returns... PMID- 18401365 TI - Chemical weapons agency shifts focus. PMID- 18401367 TI - Gertrude versus Goliath. PMID- 18401369 TI - Bill Foster. PMID- 18401370 TI - Poll results: look who's doping. PMID- 18401371 TI - Radio sweat gland--90 GHz. PMID- 18401379 TI - Evolution: scandal! Sex-starved and still surviving. PMID- 18401380 TI - Collaborations: with all good intentions. PMID- 18401381 TI - Cleaning up the final phase of the fossil-fuel industry. PMID- 18401382 TI - Italy must invest more in science and technology. PMID- 18401383 TI - CropLife still committed to assessment's original aims. PMID- 18401384 TI - Collaborations: investigating international misconduct. PMID- 18401394 TI - Rise of the digital machine. PMID- 18401395 TI - Atmospheric chemistry: are plant emissions green? PMID- 18401396 TI - Immunology: blood lines redrawn. PMID- 18401397 TI - Materials science: strong teeth, strong seeds. PMID- 18401399 TI - Quantum physics: observations turn up the heat. PMID- 18401400 TI - Cell biology: porter and sorter. PMID- 18401401 TI - Neurodegeneration: a question of balance. PMID- 18401402 TI - Cell biology: SUMO. PMID- 18401403 TI - Clathrin is a key regulator of basolateral polarity. AB - Clathrin-coated vesicles are vehicles for intracellular trafficking in all nucleated cells, from yeasts to humans. Many studies have demonstrated their essential roles in endocytosis and cellular signalling processes at the plasma membrane. By contrast, very few of their non-endocytic trafficking roles are known, the best characterized being the transport of hydrolases from the Golgi complex to the lysosome. Here we show that clathrin is required for polarity of the basolateral plasma membrane proteins in the epithelial cell line MDCK. Clathrin knockdown depolarized most basolateral proteins, by interfering with their biosynthetic delivery and recycling, but did not affect the polarity of apical proteins. Quantitative live imaging showed that chronic and acute clathrin knockdown selectively slowed down the exit of basolateral proteins from the Golgi complex, and promoted their mis-sorting into apical carrier vesicles. Our results demonstrate a broad requirement for clathrin in basolateral protein trafficking in epithelial cells. PMID- 18401404 TI - Thermodynamic control by frequent quantum measurements. AB - Heat flow between a large thermal 'bath' and a smaller system brings them progressively closer to thermal equilibrium while increasing their entropy. Fluctuations involving a small fraction of a statistical ensemble of systems interacting with the bath result in deviations from this trend. In this respect, quantum and classical thermodynamics are in agreement. Here we predict a different trend in a purely quantum mechanical setting: disturbances of thermal equilibrium between two-level systems (TLSs) and a bath, caused by frequent, brief quantum non-demolition measurements of the TLS energy states. By making the measurements increasingly frequent, we encounter first the anti-Zeno regime and then the Zeno regime (namely where the TLSs' relaxation respectively speeds up and slows down). The corresponding entropy and temperature of both the system and the bath are then found to either decrease or increase depending only on the rate of observation, contrary to the standard thermodynamical rules that hold for memory-less (Markov) baths. From a practical viewpoint, these anomalies may offer the possibility of very fast control of heat and entropy in quantum systems, allowing cooling and state purification over an interval much shorter than the time needed for thermal equilibration or for a feedback control loop. PMID- 18401405 TI - Microscopic theory of the extraordinary optical transmission. AB - The phenomenon of extraordinary light transmission through metallic films perforated by nanohole arrays at optical frequencies was first observed a decade ago and initiated important further experimental and theoretical work. In view of potential applications of such structures--for example, subwavelength optics, optoelectronics devices, and chemical sensing--it is important to understand the underlying physical processes in detail. Here we derive a microscopic theory of the transmission through subwavelength hole arrays, by considering the elementary processes associated with scattering of surface-plasmon-polariton (SPP) modes by individual one-dimensional chains of subwavelength holes. Using a SPP coupled mode model that coherently gathers these elementary processes, we derive analytical expressions for all the transmission spectrum characteristics--such as the resonance wavelength, the peak transmission and the anti-resonance. Further comparisons of the model predictions with fully vectorial computational results allow us quantitatively to check the model accuracy and to discuss the respective impacts of SPP modes and of other electromagnetic fields on producing the extraordinary transmission of light. The model greatly expands our understanding of the phenomenon and may affect further engineering of nanoplasmonic devices. PMID- 18401406 TI - Improper ferroelectricity in perovskite oxide artificial superlattices. AB - Ferroelectric thin films and superlattices are currently the subject of intensive research because of the interest they raise for technological applications and also because their properties are of fundamental scientific importance. Ferroelectric superlattices allow the tuning of the ferroelectric properties while maintaining perfect crystal structure and a coherent strain, even throughout relatively thick samples. This tuning is achieved in practice by adjusting both the strain, to enhance the polarization, and the composition, to interpolate between the properties of the combined compounds. Here we show that superlattices with very short periods possess a new form of interface coupling, based on rotational distortions, which gives rise to 'improper' ferroelectricity. These observations suggest an approach, based on interface engineering, to produce artificial materials with unique properties. By considering ferroelectric/paraelectric PbTiO3/SrTiO3 multilayers, we first show from first principles that the ground-state of the system is not purely ferroelectric but also primarily involves antiferrodistortive rotations of the oxygen atoms in a way compatible with improper ferroelectricity. We then demonstrate experimentally that, in contrast to pure PbTiO3 and SrTiO3 compounds, the multilayer system indeed behaves like a prototypical improper ferroelectric and exhibits a very large dielectric constant of epsilon(r) approximately 600, which is also fairly temperature-independent. This behaviour, of practical interest for technological applications, is distinct from that of normal ferroelectrics, for which the dielectric constant is typically large but strongly evolves around the phase transition temperature and also differs from that of previously known improper ferroelectrics that exhibit a temperature-independent but small dielectric constant only. PMID- 18401407 TI - Atmospheric oxidation capacity sustained by a tropical forest. AB - Terrestrial vegetation, especially tropical rain forest, releases vast quantities of volatile organic compounds (VOCs) to the atmosphere, which are removed by oxidation reactions and deposition of reaction products. The oxidation is mainly initiated by hydroxyl radicals (OH), primarily formed through the photodissociation of ozone. Previously it was thought that, in unpolluted air, biogenic VOCs deplete OH and reduce the atmospheric oxidation capacity. Conversely, in polluted air VOC oxidation leads to noxious oxidant build-up by the catalytic action of nitrogen oxides (NO(x) = NO + NO2). Here we report aircraft measurements of atmospheric trace gases performed over the pristine Amazon forest. Our data reveal unexpectedly high OH concentrations. We propose that natural VOC oxidation, notably of isoprene, recycles OH efficiently in low NO(x) air through reactions of organic peroxy radicals. Computations with an atmospheric chemistry model and the results of laboratory experiments suggest that an OH recycling efficiency of 40-80 per cent in isoprene oxidation may be able to explain the high OH levels we observed in the field. Although further laboratory studies are necessary to explore the chemical mechanism responsible for OH recycling in more detail, our results demonstrate that the biosphere maintains a remarkable balance with the atmospheric environment. PMID- 18401408 TI - Estimating the impact of school closure on influenza transmission from Sentinel data. AB - The threat posed by the highly pathogenic H5N1 influenza virus requires public health authorities to prepare for a human pandemic. Although pre-pandemic vaccines and antiviral drugs might significantly reduce illness rates, their stockpiling is too expensive to be practical for many countries. Consequently, alternative control strategies, based on non-pharmaceutical interventions, are a potentially attractive policy option. School closure is the measure most often considered. The high social and economic costs of closing schools for months make it an expensive and therefore controversial policy, and the current absence of quantitative data on the role of schools during influenza epidemics means there is little consensus on the probable effectiveness of school closure in reducing the impact of a pandemic. Here, from the joint analysis of surveillance data and holiday timing in France, we quantify the role of schools in influenza epidemics and predict the effect of school closure during a pandemic. We show that holidays lead to a 20-29% reduction in the rate at which influenza is transmitted to children, but that they have no detectable effect on the contact patterns of adults. Holidays prevent 16-18% of seasonal influenza cases (18-21% in children). By extrapolation, we find that prolonged school closure during a pandemic might reduce the cumulative number of cases by 13-17% (18-23% in children) and peak attack rates by up to 39-45% (47-52% in children). The impact of school closure would be reduced if it proved difficult to maintain low contact rates among children for a prolonged period. PMID- 18401409 TI - A plant pathogen virulence factor inhibits the eukaryotic proteasome by a novel mechanism. AB - Pathogenic bacteria often use effector molecules to increase virulence. In most cases, the mode of action of effectors remains unknown. Strains of Pseudomonas syringae pv. syringae (Pss) secrete syringolin A (SylA), a product of a mixed non ribosomal peptide/polyketide synthetase, in planta. Here we identify SylA as a virulence factor because a SylA-negative mutant in Pss strain B728a obtained by gene disruption was markedly less virulent on its host, Phaseolus vulgaris (bean). We show that SylA irreversibly inhibits all three catalytic activities of eukaryotic proteasomes, thus adding proteasome inhibition to the repertoire of modes of action of virulence factors. The crystal structure of the yeast proteasome in complex with SylA revealed a novel mechanism of covalent binding to the catalytic subunits. Thus, SylA defines a new class of proteasome inhibitors that includes glidobactin A (GlbA), a structurally related compound from an unknown species of the order Burkholderiales, for which we demonstrate a similar proteasome inhibition mechanism. As proteasome inhibitors are a promising class of anti-tumour agents, the discovery of a novel family of inhibitory natural products, which we refer to as syrbactins, may also have implications for the development of anti-cancer drugs. Homologues of SylA and GlbA synthetase genes are found in some other pathogenic bacteria, including the human pathogen Burkholderia pseudomallei, the causative agent of melioidosis. It is thus possible that these bacteria are capable of producing proteasome inhibitors of the syrbactin class. PMID- 18401410 TI - Endothelins are vascular-derived axonal guidance cues for developing sympathetic neurons. AB - During development, sympathetic neurons extend axons along a myriad of distinct trajectories, often consisting of arteries, to innervate one of a large variety of distinct final target tissues. Whether or not subsets of neurons within complex sympathetic ganglia are predetermined to innervate select end-organs is unknown. Here we demonstrate in mouse embryos that the endothelin family member Edn3 (ref. 1), acting through the endothelin receptor EdnrA (refs 2, 3), directs extension of axons of a subset of sympathetic neurons from the superior cervical ganglion to a preferred intermediate target, the external carotid artery, which serves as the gateway to select targets, including the salivary glands. These findings establish a previously unknown mechanism of axonal pathfinding involving vascular-derived endothelins, and have broad implications for endothelins as general mediators of axonal growth and guidance in the developing nervous system. Moreover, they suggest a model in which newborn sympathetic neurons distinguish and choose between distinct vascular trajectories to innervate their appropriate end organs. PMID- 18401411 TI - The earliest thymic progenitors for T cells possess myeloid lineage potential. AB - There exists controversy over the nature of haematopoietic progenitors of T cells. Most T cells develop in the thymus, but the lineage potential of thymus colonizing progenitors is unknown. One approach to resolving this question is to determine the lineage potentials of the earliest thymic progenitors (ETPs). Previous work has shown that ETPs possess T and natural killer lymphoid potentials, and rare subsets of ETPs also possess B lymphoid potential, suggesting an origin from lymphoid-restricted progenitor cells. However, whether ETPs also possess myeloid potential is unknown. Here we show that nearly all ETPs in adult mice possess both T and myeloid potential in clonal assays. The existence of progenitors possessing T and myeloid potential within the thymus is incompatible with the current dominant model of haematopoiesis, in which T cells are proposed to arise from lymphoid-. Our results indicate that alternative models for lineage commitment during haematopoiesis must be considered. PMID- 18401412 TI - Adult T-cell progenitors retain myeloid potential. AB - During haematopoiesis, pluripotent haematopoietic stem cells are sequentially restricted to give rise to a variety of lineage-committed progenitors. The classical model of haematopoiesis postulates that, in the first step of differentiation, the stem cell generates common myelo-erythroid progenitors and common lymphoid progenitors (CLPs). However, our previous studies in fetal mice showed that myeloid potential persists even as the lineage branches segregate towards T and B cells. We therefore proposed the 'myeloid-based' model of haematopoiesis, in which the stem cell initially generates common myelo-erythroid progenitors and common myelo-lymphoid progenitors. T-cell and B-cell progenitors subsequently arise from common myelo-lymphoid progenitors through myeloid-T and myeloid-B stages, respectively. However, it has been unclear whether this myeloid based model is also valid for adult haematopoiesis. Here we provide clonal evidence that the early cell populations in the adult thymus contain progenitors that have lost the potential to generate B cells but retain substantial macrophage potential as well as T-cell, natural killer (NK)-cell and dendritic cell potential. We also show that such T-cell progenitors can give rise to macrophages in the thymic environment in vivo. Our findings argue against the classical dichotomy model in which T cells are derived from CLPs; instead, they support the validity of the myeloid-based model for both adult and fetal haematopoiesis. PMID- 18401414 TI - Sphingosine kinase-1 is a downstream regulator of imatinib-induced apoptosis in chronic myeloid leukemia cells. AB - We examined the involvement of sphingosine kinase-1 (SphK1), which governs the ceramide/sphingosine-1-phosphate balance, in susceptibility to imatinib of either sensitive or resistant chronic myeloid leukemia cells. Imatinib-sensitive LAMA84 s displayed marked SphK1 inhibition coupled with increased content of ceramide and decreased pro-survival sphingosine-1-phosphate. Conversely, no changes in the sphingolipid metabolism were observed in LAMA84-r treated with imatinib. Overcoming imatinib resistance in LAMA84-r with farnesyltransferase or MEK/ERK inhibitors as well as with cytosine arabinoside led to SphK1 inhibition. Overexpression of SphK1 in LAMA84-s cells impaired apoptosis and inhibited the effects of imatinib on caspase-3 activation, cytochrome c and Smac release from mitochondria through modulation of Bim, Bcl-xL and Mcl-1 expression. Pharmacological inhibition of SphK1 with F-12509a or its silencing by siRNA induced apoptosis of both imatinib-sensitive and -resistant cells, suggesting that SphK1 inhibition was critical for apoptosis signaling. We also show that imatinib-sensitive and -resistant primary cells from chronic myeloid leukemia patients can be successfully killed in vitro by the F-12509a inhibitor. These results uncover the involvement of SphK1 in regulating imatinib-induced apoptosis and establish that SphK1 is a downstream effector of the Bcr-Abl/Ras/ERK pathway inhibited by imatinib but upstream regulator of Bcl-2 family members. PMID- 18401415 TI - STAT6 activity is regulated by SOCS-1 and modulates BCL-XL expression in primary mediastinal B-cell lymphoma. PMID- 18401416 TI - Dasatinib induces durable cytogenetic responses in patients with chronic myelogenous leukemia in chronic phase with resistance or intolerance to imatinib. AB - Dasatinib, a potent inhibitor of BCR-ABL in vitro, is effective for patients with chronic myelogenous leukemia (CML) resistant or intolerant to imatinib. To provide a more definitive assessment of dasatinib in chronic-phase (CP)-CML, we report extended follow-up of a phase II trial, presenting data for the entire patient cohort (N=387). Dasatinib (70 mg) twice daily was administered to patients with imatinib-resistant or -intolerant CP-CML. With median follow-up of 15.2 months (treatment duration, <1-18.4 months), a complete hematologic response was attained or maintained in 91% of patients. A major cytogenetic response (MCyR) was attained or maintained by 59% (52% imatinib resistant and 80% imatinib intolerant); this was complete in 49% of patients (40% imatinib resistant and 75% imatinib intolerant). Of 230 patients achieving an MCyR, 7 experienced disease progression. Fifteen-month progression-free survival was 90% while overall survival was 96%. Grade 3/4 thrombocytopenia and neutropenia were reported in 48 and 49% of patients, respectively. Non-hematologic toxicity (any grade) consisted primarily of diarrhea (37%), headache (32%), fatigue (31%), dyspnea (30%) and pleural effusion (27%). Pleural effusions were classified as grade 3 in 6% of reported events, with no incidence of grade 4. Dasatinib is associated with high response rates in patients with imatinib-resistant or -intolerant CP-CML. PMID- 18401417 TI - Activity of tyrosine kinase inhibitors against human NUP214-ABL1-positive T cell malignancies. AB - Amplification of the NUP214-ABL1 oncogene can be detected in patients with T cell acute lymphoblastic leukemia (T-ALL). We screened 29 patients with T cell malignancies for the expression of NUP214-ABL1 by reverse transcription polymerase chain reaction (RT-PCR). NUP214-ABL1 was detected in three (10%) patients. These results were confirmed by fluorescence in situ hybridization techniques. We also studied the activity of imatinib, nilotinib and dasatinib against the human NUP214-ABL1-positive cell lines PEER and BE-13. All three tyrosine kinase inhibitors decreased the viability of PEER and BE-13 cells, but nilotinib and dasatinib had >1-log lower IC(50) values than imatinib (P<0.001). In contrast, the NUP214-ABL-negative T-ALL cell line Jurkat, was remarkably resistant to tyrosine kinase inhibition. The inhibition of cellular proliferation was associated with time-dependent induction of apoptosis and inhibition of ABL, CrKL and STAT5 phosphorylation. Moreover, dasatinib was active in a NUP214-ABL1 positive leukemia xenograft murine model and in marrow lymphoblasts from a patient with NUP214-ABL1-positive T-ALL. On the basis of these results, ABL1 kinase inhibitors warrant clinical investigation in patients with NUP214-ABL1 positive T-cell malignancies. PMID- 18401418 TI - BCR/ABL promotes accumulation of chromosomal aberrations induced by oxidative and genotoxic stress. PMID- 18401419 TI - Overexpression of Syk tyrosine kinase in peripheral T-cell lymphomas. AB - Peripheral T-cell lymphomas (PTCLs) are fatal in the majority of patients and novel treatments, such as protein tyrosine kinase (PTK) inhibition, are needed. The recent finding of SYK/ITK translocations in rare PTCLs led us to examine the expression of Syk PTK in 141 PTCLs. Syk was positive by immunohistochemistry (IHC) in 133 PTCLs (94%), whereas normal T cells were negative. Western blot on frozen tissue (n=6) and flow cytometry on cell suspensions (n=4) correlated with IHC results in paraffin. Additionally, western blot demonstrated that Syk positive PTCLs show tyrosine (525/526) phosphorylation, known to be required for Syk activation. Fluorescence in situ hybridization showed no SYK/ITK translocation in 86 cases. Overexpression of Syk, phosphorylation of its Y525/526 residues and the availability of orally available Syk inhibitors suggest that Syk merits further evaluation as a candidate target for pharmacologic PTK inhibition in patients with PTCL. PMID- 18401420 TI - Natural T-cell responses against minor histocompatibility antigen (mHag) HY following HLA-matched hematopoietic cell transplantation: what are the requirements for a 'good' mHag? PMID- 18401421 TI - Increase sensitivity to chemotherapeutical agents and cytoplasmatic interaction between NPM leukemic mutant and NF-kappaB in AML carrying NPM1 mutations. AB - Mutations in nucleophosmin (NPM) exon 12 and the resulting delocalization of NPM into the cytoplasm are the most specific and frequent cellular events in acute myeloid leukemia patients (AML) with normal karyotype. Cytoplasmatic NPM (NPMc+) is associated with responsiveness to chemotherapy and better prognosis. The activation of nuclear factor-kappaB (NF-kappaB) has been demonstrated to occur in a subset of AML patients and is thought to induce resistance to many chemotherapeutical agents. In this study, we demonstrate the increased in vitro sensitivity of NPMc+ cells to chemotherapeutical agents and their reduced NF kappaB activity. Furthermore, we provide evidence of the interaction between NPMc+ and NF-kappaB in the cytoplasm, resulting in the sequestration and inactivation of NF-kappaB. The cytosolic localization and consequent inactivation of NF-kappaB justifies the reduced NF-kappaB DNA-binding activity observed in NPMc+ patients. These data, taken together, may provide a possible explanation for the increased rate of chemosensitivity observed among the NPMc+ patients. PMID- 18401422 TI - Hic-5 promotes the hypertrophic scar myofibroblast phenotype by regulating the TGF-beta1 autocrine loop. AB - Following severe traumatic or thermal injury to the dermis, hypertrophic scars (HTSs) often develop in humans. These scar fibroblasts (hypertrophic scar fibroblasts (HTSFs)) retain the myofibroblast phenotype persistently, rather than transiently as in acute wounds. These pathogenic myofibroblasts constitutively express smooth-muscle cell alpha-actin (SMAA), deposit an excessive amount of extracellular matrix (ECM) proteins, are highly contractile, and stably display large focal adhesions. Increasing evidence supports a mechanism in which autocrine production and activation of transforming growth factor-beta1 (TGF beta1) are required to maintain the pathogenic myofibroblast phenotype. We recently reported that Hic-5, a focal adhesion protein that is upregulated by TGF beta1, is expressed persistently in HTSF compared to normal adult fibroblasts (NADFs). We now find that Hic-5 is an important regulator of the constitutive myofibroblast phenotype in HTSFs. Silencing the expression of Hic-5 in HTSFs with specific siRNAs dramatically reduces TGF-beta1 production, decreases the generation of supermature focal adhesions reduces expression of SMAA and decreases collagen contraction and ECM synthesis. Our findings demonstrate that Hic-5 is an essential component of the mechanism regulating autocrine production of TGF-beta1 and the resulting pathogenic myofibroblast phenotype. PMID- 18401423 TI - Tie2-R849W mutant in venous malformations chronically activates a functional STAT1 to modulate gene expression. AB - Tie2 is an endothelial receptor tyrosine kinase. An amino-acid substitution of tryptophan for arginine at residue 849 (Tie2-R849W) leads to a ligand-independent activation of its kinase activity. This mutation has been associated with familial venous malformations (VMs), manifested by variable thickness or lack of smooth-muscle cells in the veins of patient lesions. The underlying mechanism for Tie2-R849W action in endothelial cells remains elusive. In this study, we used adenoviral infection to differentiate the effects of ectopic Tie2 (wild type, kinase-dead K855A, or constitutively active R849W) expression on endothelial cellular behaviors and Tie2-mediated downstream targets. Ectopic Tie2 reduced endothelial cell proliferation and serum withdrawal-induced apoptosis, while stimulating migration. When comparing R849W with K855A and its wild-type counterpart, a functional tyrosine kinase activity was required only for migration, and constitutively active Tie2-R849W conferred highest resistance to serum-induced apoptosis, but lowest ability to maintain tube-like structures formed on Matrigel. We further demonstrated that Tie2-R849W chronically induced STAT1 tyrosine phosphorylation and the promoter activity of STAT1-responsive IFN regulatory factor 1 (IRF1). Although STAT1 phosphorylation required JNK and p38MAPK activation, only JNK activation was essential for IRF1 promoter activation by Tie2-R849W. Additional studies are needed to study the role of STAT1 activation in VMs. PMID- 18401424 TI - Systemic PPARgamma ligation inhibits allergic immune response in the skin. AB - We have shown previously that specific ligands of the peroxisome proliferator activated receptor-gamma (PPARgamma) inhibit the systemic allergic immune response. The objective of this study was to investigate the impact of PPARgamma ligand treatment on the local allergic immune response. We established a murine model exhibiting clinical and histological features of AD-like skin lesions with high reproducibility. In this model, the PPARgamma ligand was applied in an either preventive or therapeutic manner via systemic and local routes. The affected skin areas were assessed by standardized skin score, histological analyses, and immunohistochemical examinations. Our data show that systemic application of PPARgamma ligand by a preventive protocol led to significantly reduced onset of eczematous skin lesions. This was confirmed by histology, showing decreased skin thickness accompanied by significantly reduced infiltrations of CD4+ and CD8+ lymphocytes but also mast cells. Additionally, early allergen-specific IgE and IgG1 responses were reduced (day 21/35), whereas IgG2a levels remained unchanged. In conclusion, our results demonstrate that PPARgamma-ligand treatment inhibits not only systemic allergic immune response, but also local allergen-mediated dermatitis. Our findings point to therapeutic strategies, including a PPARgamma-ligand-based treatment. PMID- 18401425 TI - The effects of depilatory agents as penetration enhancers on human stratum corneum structures. AB - The depilatory cream thioglycolate has been shown to be an effective enhancer for transdermal drug delivery. However the mechanism remains unknown. In addition, it may also increase the risk of permeation of exogenous toxic agents across skin in depilatory cream users. The aim of this study was to characterize its effect on the transepidermal route and the associated structural alterations. Fresh human skin was treated with a depilatory cream for 10 minutes and then permeated with fluorescent model drugs. The penetration of model drugs was then imaged and quantified. The structural alternations of stratum corneum were assessed by multi photon imaging, histology, Nile red staining, and electron microscopy. Our results show that penetration of both hydrophilic and hydrophobic model drugs across stratum corneum was enhanced. Disruption of cellular integrity and focal detachment of superficial corneocytes was observed in multi-photon imaging. In addition, nile red staining showed disorganized lipid distribution. Finally, ultrastructural analysis revealed disruption of intracellular keratin matrix, protein cell envelope, and regular lamellar intercellular lipid packing. Because intracellular and intercellular structures were altered, our results suggest that depilatory agents enhance transepidermal drug delivery by reducing resistance in both transcellular and intercellular routes of stratum corneum. PMID- 18401426 TI - Proteolytic activation of matrix metalloproteinase-9 in skin wound healing is inhibited by alpha-1-antichymotrypsin. AB - An excessive amount of matrix metalloproteinase-9 (MMP-9) has been well documented in inflammatory diseases, including chronic wounds and cancers. Secreted as a zymogen, proMMP-9 can be irreversibly converted to a mature form through cleavage of the N-terminal propeptide domain. Although the converting enzyme for proMMP-9 in human tissues is unknown, we previously found that tumor necrosis factor-alpha (TNF-alpha) promotes activation of proMMP-9 in human skin, and characterized the converting activities as tissue-associated chymotrypsin like proteinases. On the other hand, the pathophysiologic inhibitor to prevent proMMP-9 maturation also remains elusive. In this regard, we observed the presence of the inhibitory property in burn blister fluid that abrogates the skin extract-mediated activation of proMMP-9. Then we determined that alpha-1 antichymotrypsin (alpha-ACT), an acute-phase factor abundantly present in the blister, effectively inhibited proMMP-9 activation in human and rodent skin. In contrast, the aminophenylmercuric acetate-induced "cysteine switch" and activation of proMMP-9 were not affected by alpha-ACT. TNF-alpha-induced activation of proMMP-9 by the explants of human skin was inhibited by alpha-ACT but not by related alpha-1-antitrypsin. alpha-ACT specifically attenuated maturation of proMMP-9 but not proMMP-2 or proMMP-13. Furthermore, short peptides that mimic the reactive center loop (RCL) of alpha-ACT were sufficient to inhibit the conversion. Mutation analysis demonstrated that a conserved leucine within the RCL was critical for alpha-ACT-exerted inhibition. In chronic wounds, a large amount of mature MMP-9 was associated with fragmentation and inactivation of alpha-ACT. Taken together, these results demonstrate that, to the best of our knowledge, alpha-ACT is a previously unreported pathophysiologic inhibitor that controls proMMP-9 activation in skin tissue. PMID- 18401427 TI - The human papillomavirus type 8 E2 protein induces skin tumors in transgenic mice. AB - Transgenic mice expressing early genes of the cutaneous human papillomavirus 8 (HPV8) spontaneously develop skin papillomas, epidermal dysplasia, and squamous cell carcinoma (6%). As the HPV8 protein E2 revealed transforming capacity in vitro, we generated three epidermal specific HPV8-E2-transgenic FVB/N mouse lines to dissect its role in tumor development. The rate of tumor formation in the three lines correlated with the different E2-mRNA levels. More than 60% of heterozygous line 2 mice, but none of the HPV8-negative littermates, spontaneously developed ulcerous lesions of the skin over an observation period of up to 144 weeks, beginning on average 74+/-22 weeks after birth. Most lesions presented infundibular hyperplasia and acanthosis combined with low-grade dysplasia. Severe dysplasia of the epidermis occurred in 6%. Two carcinomas revealed a sharply demarcated spindle-cell component. Only 3 weeks after a single UV irradiation, 87% of heterozygous line 2 and 36% of line 35 mice developed skin tumors. A rapidly growing invasive tumor composed of spindle cells arose 10 weeks after irradiation of a line-35 animal. The histology of skin cancers in HPV8-E2 mice is reminiscent of a subset of highly aggressive squamous cell carcinoma in immunosuppressed transplant recipients with a massive spindle-cell component. PMID- 18401428 TI - Plasticity of the cancer cell: implications for epigenetic control of melanoma and other malignancies. AB - Current treatments of many advanced malignancies, including melanoma, have failed to significantly reduce mortality rates, necessitating newer approaches. There is now abundant evidence that cancer cells, given the appropriate environmental and molecular context, are capable of remarkable plasticity, including complete reversal of the malignant phenotype. Such reprogramming involves both extrinsic and intrinsic factors and can occur via three routes: perturbations of extracellular matrix-cell receptor interactions, modulation of intracellular signaling pathways, and exploitation of epigenetic inheritance. Studies demonstrate the potential for producing dramatic changes in structural, biochemical, immunological, and functional properties of a broad spectrum of tumor cell types, including melanoma, leading to growth arrest, differentiation, senescence, or self destruction. Translating the promise inherent in tumor cell plasticity to the clinical arena remains a major challenge, but it is likely that a variety of epigenetic methods will play an increasingly important and effective role in the future control of malignant melanoma and other cancers. PMID- 18401429 TI - A 3' UTR modification of the mitochondrial rieske iron sulfur protein in mice produces a specific skin pigmentation phenotype. PMID- 18401430 TI - Knockdown of myosin Va isoforms by RNAi as a tool to block melanosome transport in primary human melanocytes. AB - The movement of melanosomes, dense melanin-containing organelles, within human melanocytes is actin-dependent and mediated through the formation of a Rab27a Slac2-a-myosin Va (MyoVa) protein complex. We previously showed that only the melanocyte-specific exon F isoforms of MyoVa are involved in melanosome transport to the dendrite extremities. Here, we investigate siRNA to downregulate the exon F-containing isoforms of MyoVa in primary human melanocytes. Efficient and specific knockdown of the MyoVa exon F isofoms were shown at both mRNA and protein levels. Further, a stable shRNA against the MyoVa exon F isoforms was prepared by using a lentiviral system to improve and confirm the silencing effect in hard-to-transfect melanocyte cells. Immunofluorescence microscopy shows that knockdown of the exon F isoforms results in blockade of intramelanocytic melanosome transport due to the inability to form the Rab27a-Slac2-a-MyoVa tripartite complex. Interestingly, the observed phenotypic effect (that is, perinuclear accumulation of melanosomes) is the same as that seen in melanocytes from patients with human Griscelli syndrome causing abnormal pigmentation. We conclude that our siRNA-based strategy provides a previously unreported tool to block the intracellular melanosome movement in primary human melanocytes and may become an innovative drug to treat hyperpigmentation. PMID- 18401431 TI - Changing the adenovirus fiber for retaining gene delivery efficacy in the presence of neutralizing antibodies. AB - Prior infection has primed most adult humans for a rapid neutralizing antibody (NAb) response when re-exposed to adenovirus. NAb induction can severely limit the efficacy of systemic re-administration of adenoviral gene therapy. We hypothesized that changing the fiber knob could overcome NAb. Immune-competent mice were exposed to serotype 5 adenovirus (Ad5)(GL), Ad5/3luc1, Ad5lucRGD or Ad5pK7(GL). Mice immunized with Ad5(GL) featured reduced intravenous Ad5(GL) gene transfer to most organs, including the liver, lung and spleen. Ad5(GL) gene transfer was affected much less by exposure to capsid-modified viruses. Anti Ad5(GL) NAb blocked intravenous Ad5(GL) gene transfer to orthotopic lung cancer xenografts, whereas capsid-modified viruses were not affected. When gene transfer to fresh cancer and normal lung explants was analyzed, we found that capsid modified viruses allowed effective gene delivery to tumors in the presence of anti-Ad5(GL) NAb, whereas Ad5(GL) was blocked. In contrast, crossblocking by NAbs induced by different viruses affected gene delivery to normal human lung explants, suggesting the importance of non-fiber-knob-mediated infection mechanisms. We conclude that changing the adenovirus fiber knob is sufficient to allow a relative degree of escape from preexisting NAb. If confirmed in trials, this approach might improve the efficacy of re-administration of adenoviral gene therapy to humans. PMID- 18401432 TI - Potential of AAV vectors in the treatment of metabolic disease. AB - Inborn errors of metabolism are collectively common, frequently severe and in many instances difficult or impossible to treat. Accordingly, there is a compelling need to explore novel therapeutic modalities, including gene therapy, and examine multiple phenotypes where the risks of experimental therapy are outweighed by potential benefits to trial participants. Among available gene delivery systems recombinant AAV shows special promise for the treatment of metabolic disease given the unprecedented efficiencies achieved in transducing key target tissues, such as liver and muscle, in small animal models. To date over 30 metabolic disease phenotypes have been investigated in small animal studies with complete phenotype correction being achieved in a substantial proportion. Achieving adequately widespread transduction within the central nervous system, however, remains a major challenge, and will be critical to realization of the therapeutic potential of gene therapy for many of the most clinically troubling metabolic disease phenotypes. Despite the relatively low immunogenicity of AAV vectors, immune responses are also emerging as a factor requiring special attention as efforts accelerate toward human clinical translation. Four metabolic disease phenotypes have reached phase I or I/II trials with one, targeting lipoprotein lipase deficiency, showing exciting early evidence of efficacy. PMID- 18401433 TI - Toward exascale production of recombinant adeno-associated virus for gene transfer applications. AB - To gain acceptance as a medical treatment, adeno-associated virus (AAV) vectors require a scalable and economical production method. Recent developments indicate that recombinant AAV (rAAV) production in insect cells is compatible with current good manufacturing practice production on an industrial scale. This platform can fully support development of rAAV therapeutics from tissue culture to small animal models, to large animal models, to toxicology studies, to Phase I clinical trials and beyond. Efforts to characterize, optimize and develop insect cell based rAAV production have culminated in successful bioreactor-scale production of rAAV, with total yields potentially capable of approaching the exa-(10(18)) scale. These advances in large-scale AAV production will allow us to address specific catastrophic, intractable human diseases such as Duchenne muscular dystrophy, for which large amounts of recombinant vector are essential for successful outcome. PMID- 18401434 TI - The size of endothelial fenestrae in human liver sinusoids: implications for hepatocyte-directed gene transfer. AB - Fenestrae allow the passage of gene transfer vectors from the sinusoidal lumen to the surface of hepatocytes. We have previously shown that the diameter of fenestrae correlates with species and strain differences of transgene expression following intravenous adenoviral transfer. In the current study, we demonstrate that the diameter of fenestrae in humans without liver pathology is 107+/-1.5 nm. This is similar to the previously reported diameter in New Zealand White (NZW) rabbits (103+/-1.3 nm) and is significantly smaller than in C57BL/6 mice (141+/ 5.4 nm) and Sprague-Dawley rats (161+/-2.7 nm). We show that the diameter of fenestrae in one male NZW rabbit and its offspring characterized by a more than 50-fold increase of transgene expression after adenoviral gene transfer is significantly (113+/-1.5 nm; P<0.001) larger than in control NZW rabbits. In vitro filtration experiments using polycarbonate filters with increasing pore sizes demonstrate that a relatively small increment of the diameter of pores potently enhances passage of adenoviral vectors, consistent with in vivo data. In conclusion, the small diameter of fenestrae in humans is likely to be a major obstacle for hepatocyte transduction by adenoviral vectors. PMID- 18401436 TI - Adeno-associated virus integration: virus versus vector. AB - Although a large percentage of the world population is seropositive for exposure to various strains of adeno-associated virus (AAV), a human parvovirus, AAV has never been identified as an etiologic agent of human disease. Most likely contributing to the pronounced lack of pathogenicity is the fact that AAV is a naturally defective virus that requires a helper virus for productive replication of its genome. Another unusual aspect of wild-type AAV biology is the ability of the virus to establish latent infection by preferential integration of its genome into a specific locus of human chromosome 19. Site-specific integration was a major impetus for the development of recombinant AAV vectors, which typically lack all AAV coding sequences. It was soon realized, however, that expression of at least one species of the virally encoded initiator proteins, Rep78 or Rep68, is necessary for targeted integration of AAV-derived DNA constructs to occur. This article will present a chronological outline of studies characterizing site specific integration of wild-type AAV sequences and the quasi-random target site selection observed with recombinant AAV vectors. PMID- 18401435 TI - Inflammatory tumour cell killing by oncolytic reovirus for the treatment of melanoma. AB - Reovirus is a promising unmodified double-stranded RNA (dsRNA) anti-cancer oncolytic virus, which is thought to specifically target cells with activated Ras. Although reovirus has been tested in a wide range of preclinical models and has entered early clinical trials, it has not previously been tested for the treatment of human melanoma. Here, we show that reovirus effectively kills and replicates in both human melanoma cell lines and freshly resected tumour; intratumoural injection also causes regression of melanoma in a xenograft in vivo model. Reovirus-induced melanoma death is blocked by caspase inhibition and is dependent on constituents of the Ras/RalGEF/p38 pathway. Reovirus melanoma killing is more potent than, and distinct from, chemotherapy or radiotherapy induced cell death; a range of inflammatory cytokines and chemokines are released by infected tumour cells, while IL-10 secretion is abrogated. Furthermore, the inflammatory response generated by reovirus-infected tumour cells causes bystander toxicity against reovirus-resistant tumour cells and activates human myeloid dendritic cells (DC) in vitro. Hence, reovirus is suitable for clinical testing in melanoma, and may provide a useful danger signal to reverse the immunologically suppressive environment characteristic of this tumour. PMID- 18401437 TI - Cluster intradermal DNA vaccination rapidly induces E7-specific CD8+ T-cell immune responses leading to therapeutic antitumor effects. AB - Intradermal administration of DNA vaccines via a gene gun represents a feasible strategy to deliver DNA directly into the professional antigen-presenting cells (APCs) in the skin. This helps to facilitate the enhancement of DNA vaccine potency via strategies that modify the properties of APCs. We have previously demonstrated that DNA vaccines encoding human papillomavirus type 16 (HPV-16) E7 antigen linked to calreticulin (CRT) are capable of enhancing the E7-specific CD+ T-cell immune responses and antitumor effects against E7-expressing tumors. It has also been shown that cluster (short-interval) DNA vaccination regimen generates potent immune responses in a minimal time frame. Thus, in the current study we hypothesize that the cluster intradermal CRT/E7 DNA vaccination will generate significant antigen-specific CD8+ T-cell infiltrates in E7-expressing tumors in tumor-bearing mice, leading to an increase in apoptotic tumor cell death. We found that cluster intradermal CRT/E7 DNA vaccination is capable of rapidly generating a significant number of E7-specific CD8+ T cells, resulting in significant therapeutic antitumor effects in vaccinated mice. We also observed that cluster intradermal CRT/E7 DNA vaccination in the presence of tumor generates significantly higher E7-specific CD8+ T-cell immune responses in the systemic circulation as well as in the tumors. In addition, this vaccination regimen also led to significantly lower levels of CD4+Foxp3+ T-regulatory cells and myeloid suppressor cells compared to vaccination with CRT DNA in peripheral blood and in tumor-infiltrating lymphocytes, resulting in an increase in apoptotic tumor cell death. Thus, our study has significant potential for future clinical translation. PMID- 18401438 TI - Inflammation and tumor microenvironments: defining the migratory itinerary of mesenchymal stem cells. AB - Mesenchymal stem cells (MSC) exhibit tropism for sites of tissue damage as well as the tumor microenvironment. Many of the same inflammatory mediators that are secreted by wounds are found in the tumor microenvironment and are thought to be involved in attracting MSC to these sites. Cell migration is dependent on a multitude of signals ranging from growth factors to chemokines secreted by injured cells and/or respondent immune cells. MSC are likely to have chemotactic properties similar to other immune cells that respond to injury and sites of inflammation. Thus, the well-described model of leukocyte migration can serve as a reasonable example to facilitate the identification of factors involved in MSC migration. Understanding the factors involved in regulating MSC migration to tumors is essential to ultimately develop novel clinical strategies aimed at using MSC as vehicles to deliver antitumor proteins or suppress MSC migration to reduce tumor growth. For example, radiation enhances inflammatory signaling in the tumor microenvironment and may be used to potentiate site-specific MSC migration. Alternatively, restricting the migration of the MSC to the tumor microenvironment may prevent competent tumor-stroma formation, thereby hindering the growth of the tumor. In this review, we will discuss the role of inflammatory signaling in attracting MSC to tumors. PMID- 18401440 TI - Resonance wavelength-dependent signal of absorptive particles in surface plasmon resonance-based detection. AB - We describe the resonance wavelength-dependent signal of absorptive particles in surface plasmon resonance (SPR)-based detection using both modeling and experimental results. The particles, gold nanocages, have a significant absorption cross-section in the nearinfrared (NIR), resulting in a wavelength dependent refractive index as measured by SPR. The SPR signal due to the nanocages varies by 4-fold over resonance wavelengths from 650 nm to 950 nm. The greatest SPR signal occurs at the longest resonance wavelengths; its magnitude is due to the inherent increase in sensitivity of SPR on gold with increasing wavelength and the optical absorption properties of the nanocages. PMID- 18401439 TI - Molecular analysis of the gut microbiota of identical twins with Crohn's disease. AB - Increasing evidence suggests that a combination of host genetics and the composition of the gut microbiota are important for development of Crohn's disease (CD). Our aim was to study identical twins with CD to determine microbial factors independent of host genetics. Fecal samples were studied from 10 monozygotic twin pairs with CD (discordant n=6 and concordant n=4) and 8 healthy twin pairs. DNA was extracted, 16S rRNA genes were PCR amplified and T-RFLP fingerprints generated using general bacterial and Bacteroides group-specific primers. The microbial communities were also profiled based on their percentage G+C contents. Bacteroides 16S rRNA genes were cloned and sequenced from a subset of the samples. The bacterial diversity in each sample and similarity indices between samples were estimated based on the T-RFLP data using a combination of statistical approaches. Healthy individuals had a significantly higher bacterial diversity compared to individuals with CD. The fecal microbial communities were more similar between healthy twins than between twins with CD, especially when these were discordant for the disease. The microbial community profiles of individuals with ileal CD were significantly different from healthy individuals and those with colonic CD. Also, CD individuals had a lower relative abundance of B. uniformis and higher relative abundances of B. ovatus and B. vulgatus. Our results suggest that genetics and/or environmental exposure during childhood, in part, determine the gut microbial composition. However, CD is associated with dramatic changes in the gut microbiota and this was particularly evident for individuals with ileal CD. PMID- 18401441 TI - Fluorescence resonance energy transfer in quantum dot-protein kinase assemblies. AB - In search of viable strategies to identify selective inhibitors of protein kinases, we have designed a binding assay to probe the interactions of human phosphoinositide-dependent protein kinase-1 (PDK1) with potential ligands. Our protocol is based on fluorescence resonance energy transfer (FRET) between semiconductor quantum dots (QDs) and organic dyes. Specifically, we have expressed and purified the catalytic kinase domain of PDK1 with an N-terminal histidine tag [His(6)-PDK1(DeltaPH)]. We have conjugated this construct to CdSe ZnS core-shell QDs coated with dihydrolipoic acid (DHLA) and tested the response of the resulting assembly to a molecular dyad incorporating an ATP ligand and a BODIPY chromophore. The supramolecular association of the BODIPY-ATP dyad with the His(6)-PDK1(DeltaPH)-QD assembly encourages the transfer of energy from the QDs to the BODIPY dyes upon excitation. The addition of ATP results in the displacement of BODIPY-ATP from the binding domain of the His(6)-PDK1(DeltaPH) conjugated to the nanoparticles. The competitive binding, however, does not prevent the energy transfer process. A control experiment with QDs, lacking the His(6)-PDK1(DeltaPH), indicates that the BODIPY-ATP dyad adsorbs nonspecifically on the surface of the nanoparticles, promoting the transfer of energy from the CdSe core to the adsorbed BODIPY dyes. Thus, the implementation of FRET-based assays to probe the binding domain of PDK1 with luminescent QDs requires the identification of energy acceptors unable to interact nonspecifically with the surface of the nanoparticles. PMID- 18401442 TI - Competition of Lactobacillus paracasei with Salmonella enterica for adhesion to Caco-2 cells. AB - Competition of commensal and probiotic bacteria with pathogens for adhesion and colonization is one of the important protective mechanisms of gastrointestinal tract. In this study, we examined the ability of Lactobacillus paracasei to inhibit the adhesion of pathogenic Salmonella enterica to human colon adenocarcinoma Caco-2 cells. Caco-2 cells were grown for 6 or 21 days to obtain nondifferentiated or well-differentiated cells, respectively. In adhesion experiments, bacteria were added to the cells for 2 or 4 hours. The number of attached bacteria was expressed as colony-forming units (CFUs), Caco-2 cells were counted in hematocytometer. Both bacterial strains used adhered better to well differentiated than to nondifferentiated Caco-2 cells, however, the amount of Salmonella adhered to Caco-2 after 2 hours of contact was 12-fold higher in comparison to L. paracasei and almost 27-fold higher after 4 hours of contact. Two types of experiments were done: coincubation (both bacteria were added to Caco-2 cells simultaneously), and preincubation (L. paracasei was incubated with Caco-2 cells first, and then S. enterica was added). In coincubation experiment, the presence of L. paracasei decreased S. enterica adhesion by 4-fold and in preincubation experiment even 7-fold. Generally, Lactobacillus spent culture supernatants (SCSs) acted weaker as inhibitors of Salmonella adhesion in comparison to the whole L. paracasei culture in coincubation experiment. In conclusion, the displacement of pathogens by lactic acid bacteria and its secretions showed here depends on the time of bacteria-epithelial cell contact, and also on the stage of Caco-2 differentiation. PMID- 18401443 TI - Fluorescence intensity and intermittency as tools for following dopamine bioconjugate processing in living cells. AB - CdSe/ZnS quantum dots (QDs) conjugated to biomolecules that quench their fluorescence, particularly dopamine, have particular spectral properties that allow determination of the number of conjugates per particle, namely, photoenhancement and photobleaching. In this work, we quantify these properties on a single-particle and ensemble basis in order to evaluate their usefulness as a tool for indicating QD uptake, breakdown, and processing in living cells. This creates a general framework for the use of fluorescence quenching and intermittency to better understand nanoparticle-cell interactions. PMID- 18401444 TI - Potential Therapeutic Targets for PPARgamma after Spinal Cord Injury. AB - Traumatic injury to the spinal cord results in multiple anatomical, physiological, and functional deficits as a result of local neuronal and glial cell death as well as loss of descending and ascending axons traversing the injury site. The many different mechanisms thought to contribute to protracted secondary cell death and dysfunction after spinal cord injury (SCI) are potential therapeutic targets. Agents that bind and activate the transcription factor peroxisome proliferator-activated receptor-gamma (PPAR-gamma) show great promise for minimizing or preventing these deleterious cascades in other models of CNS disorders. This review will summarize the major secondary injury cascades occurring after SCI and discuss data from experimental CNS injury and disease models showing the exciting potential for PPARgamma therapies after SCI. PMID- 18401445 TI - The in vitro antitumour activity of novel, mitochondrial-interactive, gold-based lipophilic cations. AB - In this study we compared the effects of two previously described antimitochondrial gold complexes, that is, [A] [Au(dppe)(2)]Cl and [B] [Au(d4pype)(2)]Cl with two novel lipophilic cations, that is, [C] [Au(dpmaaH(2))(dpmaaSnMe(2))]Cl and [D] [Au(dpmaaSnMe(2))(2)]Cl as antimitochondrial agents. The results of this study indicate that [C] and [D] have intermediate partition coefficients and exhibited a selective uptake by cells. They exhibited a higher selectivity for the various cell lines than [A] but were more cytotoxic than [B]. There is a significant correlation between the cytotoxic potential of [A], [B], [C], and [D] and their octanol/water partition coefficients in both MCF-7 (breast cancer) and MCF-12A (nonmalignant breast) cells, whereas their cytotoxic potential and ability to induce the release of cytochrome c correlated only in the case of the MCF-12A cells. Complexes [C] and [D] are promising new chemotherapeutic drugs. These compounds target the mitochondrial membranes of certain cancer cells exploiting the differences between the mitochondrial membrane potential of these cells and normal cells. Although the concentrations of these compounds necessary to eradicate cancer cells are very high, the results provide a basis for the synthesis of a new family of compounds with intermediate partition coefficients compared to [A] and [B] but with increased activity against cancer cells. PMID- 18401446 TI - C-Peptide and atherogenesis: C-Peptide as a mediator of lesion development in patients with type 2 diabetes mellitus? AB - Patients with insulin resistance and early type 2 diabetes exhibit an increased propensity to develop a diffuse and extensive pattern of arteriosclerosis. Typically, these patients show increased levels of C-peptide and over the last years various groups examined the effect of C-peptide in vascular cells as well as its potential role in lesion development. While some studies demonstrated beneficial effects of C-peptide, for example, by showing an inhibition of smooth muscle cell proliferation, others suggested proatherogenic mechanisms in patients with type 2 diabetes. Among them, C-peptide may facilitate the recruitment of inflammatory cells into early lesions and promote lesion progression by inducing smooth muscle cell proliferation. The following review will summarize the effects of C-peptide in vascular cells and discuss the potential role of C-peptide in atherogenesis in patients with type 2 diabetes. PMID- 18401447 TI - Building application-related patient identifiers: what solution for a European country? AB - We propose a method utilizing a derived social security number with the same reliability as the social security number. We show the anonymity techniques classically based on unidirectional hash functions (such as the secure hash algorithm (SHA-2) function that can guarantee the security, quality, and reliability of information if these techniques are applied to the Social Security Number). Hashing produces a strictly anonymous code that is always the same for a given individual, and thus enables patient data to be linked. Different solutions are developed and proposed in this article. Hashing the social security number will make it possible to link the information in the personal medical file to other national health information sources with the aim of completing or validating the personal medical record or conducting epidemiological and clinical research. This data linkage would meet the anonymous data requirements of the European directive on data protection. PMID- 18401449 TI - Increasing Efficiency and Quality by Consolidation of Clinical Chemistry and Immunochemistry Systems with MODULAR ANALYTICS SWA. AB - MODULAR ANALYTICS Serum Work Area (in USA Integrated MODULAR ANALYTICS, MODULAR ANALYTICS is a trademark of a member of the Roche Group) represents a further approach to automation in the laboratory medicine. This instrument combines previously introduced modular systems for the clinical chemistry and immunochemistry laboratory and allows customised combinations for various laboratory workloads. Functionality, practicability, and workflow behaviour of MODULAR ANALYTICS Serum Work Area were evaluated in an international multicenter study at six laboratories. Across all experiments, 236000 results from 32400 samples were generated using 93 methods. Simulated routine testing which included provocation incidents and anomalous situations demonstrated good performance and full functionality. Heterogeneous immunoassays, performed on the E-module with the electrochemiluminescence technology, showed reproducibility at the same level of the general chemistry tests, which was well within the clinical demands. Sample carryover cannot occur due to intelligent sample processing. Workflow experiments for the various module combinations, with menus of about 50 assays, yielded mean sample processing times of <38 minutes for combined clinical chemistry and immunochemistry requests; <50 minutes including automatically repeated samples. MODULAR ANALYTICS Serum Work Area offered simplified workflow by combining various laboratory segments. It increased efficiency while maintaining or even improving quality of laboratory processes. PMID- 18401448 TI - PPAR Genomics and Pharmacogenomics: Implications for Cardiovascular Disease. AB - The peroxisome proliferator-activated receptors (PPARs) consist of three related transcription factors that serve to regulate a number of cellular processes that are central to cardiovascular health and disease. Numerous pharmacologic studies have assessed the effects of specific PPAR agonists in clinical trials and have provided insight into the clinical effects of these genes while genetic studies have demonstrated clinical associations between PPAR polymorphisms and abnormal cardiovascular phenotypes. With the abundance of data available from these studies as a background, PPAR pharmacogenetics has become a promising and rapidly advancing field. This review focuses on summarizing the current state of understanding of PPAR genetics and pharmacogenetics and the important implications for the individualization of therapy for patients with cardiovascular diseases. PMID- 18401451 TI - PPARs and RXRs in Male and Female Fertility and Reproduction. PMID- 18401450 TI - CFP and YFP, but not GFP, provide stable fluorescent marking of rat hepatic adult stem cells. AB - The stable expression of reporter genes in adult stem cells (ASCs) has important applications in stem cell biology. The ability to integrate a noncytotoxic, fluorescent reporter gene into the genome of ASCs with the capability to track ASCs and their progeny is particularly desirable for transplantation studies. The use of fluorescent proteins has greatly aided the investigations of protein and cell function on short-time scales. In contrast, the obtainment of stably expressing cell strains with low variability in expression for studies on longer time scales is often problematic. We show that this difficulty is partly due to the cytotoxicity of a commonly used reporter, green fluorescent protein (GFP). To avoid GFP-specific toxicity effects during attempts to stably mark a rat hepatic ASC strain and, therefore, obtain stable, long-term fluorescent ASCs, we evaluated cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP), in addition to GFP. Although we were unable to derive stable GFP-expressing strains, stable fluorescent clones (up to 140 doublings) expressing either CFP or YFP were established. When fluorescently marked ASCs were induced to produce differentiated progeny cells, stable fluorescence expression was maintained. This property is essential for studies that track fluorescently marked ASCs and their differentiated progeny in transplantation studies. PMID- 18401452 TI - Gel electrophoresis of gold-DNA nanoconjugates. AB - Gold-DNA conjugates were investigated in detail by a comprehensive gel electrophoresis study based on 1200 gels. A controlled number of single-stranded DNA of different length was attached specifically via thiol-Au bonds to phosphine stabilized colloidal gold nanoparticles. Alternatively, the surface of the gold particles was saturated with single stranded DNA of different length either specifically via thiol-Au bonds or by nonspecific adsorption. From the experimentally determined electrophoretic mobilities, estimates for the effective diameters of the gold-DNA conjugates were derived by applying two different data treatment approaches. The first method is based on making a calibration curve for the relation between effective diameters and mobilities with gold nanoparticles of known diameter. The second method is based on Ferguson analysis which uses gold nanoparticles of known diameter as reference database. Our study shows that effective diameters derived from gel electrophoresis measurements are affected with a high error bar as the determined values strongly depend on the method of evaluation, though relative changes in size upon binding of molecules can be detected with high precision. Furthermore, in this study, the specific attachment of DNA via gold-thiol bonds to Au nanoparticles is compared to nonspecific adsorption of DNA. Also, the maximum number of DNA molecules that can be bound per particle was determined. PMID- 18401453 TI - Protein alterations in infiltrating ductal carcinomas of the breast as detected by nonequilibrium pH gradient electrophoresis and mass spectrometry. AB - Improvement of breast-cancer detection through the identification of potential cancer biomarkers is considered as a promising strategy for effective assessment of the disease. The current study has used nonequilibrium pH gradient electrophoresis with subsequent analysis by mass spectrometry to identify protein alterations in invasive ductal carcinomas of the breast from Tunisian women. We have identified multiple protein alterations in tumor tissues that were picked, processed, and unambiguously assigned identities by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF). The proteins identified span a wide range of functions and are believed to have potential clinical applications as cancer biomarkers. They include glycolytic enzymes, molecular chaperones, cytoskeletal-related proteins, antioxydant enzymes, and immunologic related proteins. Among these proteins, enolase 1, phosphoglycerate kinase 1, deoxyhemoglobin, Mn-superoxyde dismutase, alpha-B-crystallin, HSP27, Raf kinase inhibitor protein, heterogeneous nuclear ribonucleoprotein A2/B1, cofilin 1, and peptidylprolyl isomerase A were overexpressed in tumors compared with normal tissues. In contrast, the IGHG1 protein, the complement C3 component C3c, which are two newly identified protein markers, were downregulated in IDCA tissues. PMID- 18401454 TI - PPAR-alpha Ligands as Potential Therapeutic Agents for Wet Age-Related Macular Degeneration. AB - The peroxisome proliferator-activated receptors (PPAR's) are members of the steroid/thyroid nuclear receptor, superfamily of transcription factors. There are currently three known PPAR subtypes, alpha, beta, and gamma. The PPARs are now recognized participants in a number of biological pathways some of which are implicated in the pathogenesis of age-related macular degeneration (AMD). These include immune modulation, lipid regulation, and oxidant/antioxidant pathways important to the onset and progression of "dry" AMD, and vascular endothelial growth factor (VEGF) mediated pathways that stimulate choroidal neovascularization (CNV), characteristic of "wet" AMD. PPAR-alpha is found in retina and also on vascular cells important to formation of CNV. At this time, however, relatively little is known about potential contributions of PPAR-alpha to the pathogenesis of dry and wet AMD. This review examines current literature for potential roles of PPAR-alpha in the pathogenesis and potential treatment of AMD with emphasis on prevention and treatment of wet AMD. PMID- 18401455 TI - The Leishmania HSP20 is antigenic during natural infections, but, as DNA vaccine, it does not protect BALB/c mice against experimental L. amazonensis infection. AB - Protozoa of the genus Leishmania are causative agents of leishmaniasis, an important health problem in both human and veterinary medicine. Here, we describe a new heat shock protein (HSP) in Leishmania, belonging to the small HSP (sHSP) family in kinetoplastids. The protein is highly conserved in different Leishmania species, showing instead significant divergence with sHSP's from other organisms. The humoral response elicited against this protein during Leishmania infection has been investigated in natural infected humans and dogs, and in experimentally infected hamsters. Leishmania HSP20 is a prominent antigen for canine hosts; on the contrary, the protein seems to be a poor antigen for human immune system. Time-course analysis of appearance of anti-HSP20 antibodies in golden hamsters indicated that these antibodies are produced at late stages of the infection, when clinical symptoms of disease are patent. Finally, the protective efficacy of HSP20 was assessed in mice using a DNA vaccine approach prior to challenge with Leishmania amazonensis. PMID- 18401456 TI - Crystal structure and antitumor activity of the novel zwitterionic complex of tri n-butyltin(IV) with 2-thiobarbituric acid. AB - A novel tri-n-butyl(IV) derivative of 2-thiobarbituric acid (HTBA) of formula [(n Bu)(3)Sn(TBA) H(2)O] (1) has been synthesized and characterized by elemental analysis and (119)Sn-NMR and FT-IR spectroscopic techniques. The crystal structure of complex 1 has been determined by single crystal X-ray diffraction analysis at 120(2) K. The geometry around Sn(IV) is trigonal bipyramidal. Three n butyl groups and one oxygen atom from a deprotonated 2-thiobarbituric ligand are bonded to the metal center. The geometry is completed with one oxygen from a water molecule. Compound 1 exhibits potent, in vitro, cytotoxicity against sarcoma cancer cells (mesenchymal tissue) from the Wistar rat, polycyclic aromatic hydrocarbons (PAH, benzo[a]pyrene) carcinogenesis. In addition, the inhibition caused by 1, in the rate of lipoxygenase (LOX) catalyzed oxidation reaction of linoleic acid to hyperoxolinoleic acid, has been also kinetically and theoretically studied. The results are compared to that of cisplatin. PMID- 18401457 TI - Connectivity-based parcellation of the cortical mantle using q-ball diffusion imaging. AB - This paper exploits the idea that each individual brain region has a specific connection profile to create parcellations of the cortical mantle using MR diffusion imaging. The parcellation is performed in two steps. First, the cortical mantle is split at a macroscopic level into 36 large gyri using a sulcus recognition system. Then, for each voxel of the cortex, a connection profile is computed using a probabilistic tractography framework. The tractography is performed from q fields using regularized particle trajectories. Fiber ODF are inferred from the q-balls using a sharpening process focusing the weight around the q-ball local maxima. A sophisticated mask of propagation computed from a T1 weighted image perfectly aligned with the diffusion data prevents the particles from crossing the cortical folds. During propagation, the particles father child particles in order to improve the sampling of the long fascicles. For each voxel, intersection of the particle trajectories with the gyri lead to a connectivity profile made up of only 36 connection strengths. These profiles are clustered on a gyrus by gyrus basis using a K-means approach including spatial regularization. The reproducibility of the results is studied for three subjects using spatial normalization. PMID- 18401458 TI - AGEs and glucose levels modulate type I and III procollagen mRNA synthesis in dermal fibroblasts cells culture. AB - In the dermis, fibroblasts play an important role in the turnover of the dermal extracellular matrix. Collagen I and III, the most important dermal proteins of the extracellular matrix, are progressively altered during ageing and diabetes. For mimicking diabetic conditions, the cultured human dermal fibroblasts were incubated with increasing amounts of AGE-modified BSA and D-glucose for 24 hours. The expression of procollagen alpha2(I) and procollagen alpha1(III) mRNA was analyzed by quantitative real-time PCR. Our data revealed that the treatment of fibroblasts with AGE-modified BSA upregulated the expression of procollagen alpha2(I) and procollagen alpha1(III) mRNA in a dose-dependent manner. High glucose levels mildly induced a profibrogenic pattern, increasing the procollagen alpha2(I) mRNA expression whereas there was a downregulation tendency of procollagen alpha1(III) mRNA. PMID- 18401460 TI - X-ray phase-contrast imaging with three 2D gratings. AB - X-ray imaging is of paramount importance for clinical and preclinical imaging but it is fundamentally restricted by the attenuation-based contrast mechanism, which has remained essentially the same since Roentgen's discovery a century ago. Recently, based on the Talbot effect, groundbreaking work was reported using 1D gratings for X-ray phase-contrast imaging with a hospital-grade X-ray tube instead of a synchrotron or microfocused source. In this paper, we report an extension using 2D gratings that reduces the imaging time and increases the accuracy and robustness of phase retrieval compared to current grating-based phase-contrast techniques. Feasibility is demonstrated via numerical simulation. PMID- 18401459 TI - PPARs and Female Reproduction: Evidence from Genetically Manipulated Mice. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-activated nuclear receptors controlling many important physiological processes, including lipid and glucose metabolism, energy homeostasis, inflammation, as well as cell proliferation and differentiation. In the past decade, intensive study of PPARs has shed novel insight into prevention and treatment of dyslipidemia, insulin resistance, and type 2 diabetes. Recently, a large body of research revealed that PPARs are also functionally expressed in reproductive organs and various parts of placenta during pregnancy, which strongly suggests that PPARs might play a critical role in reproduction and development, in addition to their central actions in energy homeostasis. In this review, we summarize recent findings elucidating the role of PPARs in female reproduction, with particular focus on evidence from gene knockout and transgenic animal model study. PMID- 18401461 TI - Anti-chaperone betaA3/A1(102-117) peptide interacting sites in human alphaB crystallin. AB - PURPOSE: Our previous work identified 23 low molecular weight (<3.5 kDa) crystallin peptides in the urea-soluble fractions of normal young, normal aged, and aged cataract human lenses. We found that one of these crystallin fragments, betaA3/A1(102-117) peptide (SDAYHIERLMSFRPIC), that are present in aged and cataract lens, increased the scattering of light by beta- and gamma-crystallins and alcohol dehydrogenase (ADH) and also reduced the chaperone-like activity of alphaB-crystallin. The present study was performed to identify the interacting sites of the betaA3/A1(102-117) peptide in alphaB-crystallin. METHODS: betaA3/A1(102-117) peptide was first derivatized with sulfo-succinimidyl-2-[6 (biotinamido)-2-{p-azidobenzamido}-hexanoamido] ethyl-1-3 dithio propionate (sulfo-SBED), a photoactivable, heterotrifunctional biotin-containing cross linker. The biotin-derivatized peptide was then incubated with alphaB-crystallin at 37 degrees C for 2 h to allow complex formation followed by photolysis to facilitate the transfer of the biotin label from the peptide to alphaB crystallin. Label transfer was confirmed by western blot, and the labeled alphaB crystallin was digested with trypsin. Tryptic peptides from alphaB-crystallin carrying the biotin label were purified by avidin affinity chromatography, and betaA3/A1(102-117) peptide interacting sites in alphaB-crystallin were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and nanospray quadrupole time-of-flight mass spectrometry (QqTOF MS/MS). RESULTS: We found that the betaA3/A1(102-117) peptide interacted with alphaB-crystallin regions (70)LEKDR(74), (83)HFSPEELKVK(92), (91)VKVLGDVIEVHGK(103), (93)VLGDVIEVHGKHEER(107), and (121)KYR(123), which are part of the alpha-crystallin domain, and were previously shown to be part of the functional chaperone site in alphaB-crystallin. The betaA3/A1(102-117) peptide also interacted with regions at the COOH-terminal extension of alphaB-crystallin, (150)KQVSGPER(157), (164)EEKPAVTAAPK(174), and (164)EEKPAVTAAPKK(175). When two of the hydrophobic residues of betaA3/A1(102-117) peptide were replaced with hydrophilic residues, the resulting substituted peptide, SDADHGERLMSFRPIC, did not show the anti-chaperone property. CONCLUSIONS: This study confirmed the interactions between a low molecular weight peptide derived from betaA3/A1 crystallin found in aged and cataract lenses and alphaB-crystallin. The binding of betaA3/A1(102-117) peptide to the chaperone site and the COOH-terminal extension of alphaB-crystallin may explain its anti-chaperone property. PMID- 18401462 TI - The Governance of Genomic Information: Will it Come of Age? AB - The completion of the Human Genome Project has opened up unprecedented possibilities in healthcare, but also ethical and social dilemmas in terms of how these can be achieved. Genomic information can be seen as a "global public good" (GPG), in that it is represented by knowledge in the public domain and across national boundaries. Lack of investment, infrastructure and expertise in developing countries means that they are unable to take advantage of these GPG characteristics to address their health needs, fuelling fears of a growing "genomics divide". Some have suggested an international knowledge sharing and capacity building network, a Global Genomics Initiative, as a means to harness the potential of genomics to reduce inequalities in health between North and South. Three UNESCO declarations also call for cooperation between developed and developing countries in genomics research and science and technology in general. Using international relations theories around global governance and networks as a conceptual framework, this paper examines whether these initiatives are likely to succeed in providing effective governance of genomics. PMID- 18401464 TI - IL-10 from CD4CD25Foxp3CD127 adaptive regulatory T cells modulates parasite clearance and pathology during malaria infection. AB - The outcome of malaria infection is determined, in part, by the balance of pro inflammatory and regulatory immune responses. Failure to develop an effective pro inflammatory response can lead to unrestricted parasite replication, whilst failure to regulate this response leads to the development of severe immunopathology. IL-10 and TGF-beta are known to be important components of the regulatory response, but the cellular source of these cytokines is still unknown. Here we have examined the role of natural and adaptive regulatory T cells in the control of malaria infection and find that classical CD4+CD25(hi) (and Foxp3+) regulatory T cells do not significantly influence the outcome of infections with the lethal (17XL) strain of Plasmodium yoelii (PyL). In contrast, we find that adaptive IL-10-producing, CD4+ T cells (which are CD25-, Foxp3-, and CD127- and do not produce Th1, Th2, or Th17 associated cytokines) that are generated during both PyL and non-lethal P. yoelii 17X (PyNL) infections are able to down-regulate pro-inflammatory responses and impede parasite clearance. In summary, we have identified a population of induced Foxp3- regulatory (Tr1) T cells, characterised by production of IL-10 and down regulation of IL-7Ralpha, that modulates the inflammatory response to malaria. PMID- 18401465 TI - Determining the cost-effectiveness of preventing cardiovascular disease: are estimates calculated over the duration of a clinical trial adequate? AB - BACKGROUND: Economic analyses of randomized clinical trials often focus only on the results that are observed during the study. However, for many preventive interventions, associated costs and benefits will accrue over a patient's remaining lifetime. To determine the importance of the chosen time horizon, the cost-effectiveness (C/E) of ramipril therapy was calculated and compared in the Heart Outcomes Prevention Evaluation (HOPE), the Microalbuminuria, Cardiovascular, and Renal Outcomes in HOPE (MICRO-HOPE) and the Acute Infarction Ramipril Efficacy (AIRE) study versus the entire life expectancy (L/E) of potential patients. METHODS: The Cardiovascular Disease Life Expectancy model, a validated Markov model, was calibrated to accurately forecast the results of each trial. These results were then extrapolated over the remaining L/E of hypothetical patients 55 to 75 years of age. The predicted change in L/E and associated direct health care costs for Canadians were calculated and discounted 3% annually. RESULTS: In HOPE, the forecasted increased L/E averaged 0.06 years during the five-year study versus 1.3 years over the remaining years of L/E. The associated C/E of ramipril was $15,000 per year of life saved (YOLS) over the study duration and $8,500/YOLS over the remaining lifetime. For hypothetical patients, the C/E of ramipril over 4.5 years ranged from $6,700/YOLS to more than $58,300/YOLS and was lowest among elderly men. When the remaining L/E was considered, the C/E of ramipril was similar for men and women of all ages, ranging from $8,100/YOLS to $10,200/YOLS. The analyses of MICRO-HOPE and AIRE provided similar results. CONCLUSION: The estimated efficacy and associated C/E of ramipril in HOPE, MICRO-HOPE and the AIRE study is extremely sensitive to the selected time horizon. Economic analyses beyond the duration of randomized clinical trials are required to fully evaluate the potential costs and benefits of long-term preventive therapies. PMID- 18401466 TI - Choices, persistence and adherence to antihypertensive agents: evidence from RAMQ data. AB - BACKGROUND: Most treatment recommendations for hypertension are based on criteria that consider efficacy, safety and cost. Given the need for long-term use of antihypertensive agents, treatment compliance should also be taken into consideration in the selection process. OBJECTIVE: The purpose of the present study was to estimate persistence and adherence to antihypertensive agents in a real-life setting. METHODS: Persistence and adherence to treatment were estimated using data from the Regie de l'assurance maladie du Quebec. RESULTS: Data from a random sample of 4561 subjects with a diagnosis of hypertension covered by the Regie de l'assurance maladie du Quebec drug plan and using one of the antihypertensive agents reimbursed by the drug plan for the first time between January 2000 and December 2001 were analyzed. The persistence rate observed after a two-year period with diuretics was significantly lower (52.8%) than with any other classes of antihypertensive agent (P<0.01). Persistence rates for beta blockers, calcium channel blockers, angiotensin-II receptor blockers and angiotensin-I converting enzyme inhibitors were 69.3%, 64.3%, 60.9% and 58.9%, respectively. After two years, the proportion of patients who were 80% adherent to their treatment was 64.9% for angiotensin-I converting enzyme inhibitors, 65.0% for angiotensin-II receptor blockers, 64.2% for calcium channel blockers, 60.3% for beta-blockers and 50.9% for diuretics. The proportion of patients who were 80% adherent to their treatment was significantly lower for diuretics than with any other antihypertensive agents (P<0.01). CONCLUSION: Persistence and adherence to treatment are essential to treatment success. Results of the present study indicate that, in a real-life setting, patients are significantly less compliant to diuretics than to any other antihypertensive agents. PMID- 18401467 TI - Muscle sympathetic nerve activity and ventilation during exercise in subjects with and without chronic heart failure. AB - BACKGROUND: Changes within skeletal muscle, including augmentation of its capacity to elicit reflex increases in both efferent muscle sympathetic nerve activity (MSNA) and ventilation during work, contribute significantly to exercise intolerance in heart failure (HF). Previously, we demonstrated that peak oxygen uptake (pVO(2)) in HF relates inversely to MSNA at rest and during exercise. OBJECTIVE: To test the hypothesis that there is an independent positive relationship between resting MSNA and the ratio of ventilation to carbon dioxide output during exercise (VE/VCO(2)) that is augmented in HF. METHODS: MSNA at rest and VE/VCO(2)) during stationary cycling were measured in 30 patients (27 men) with HF (mean +/- SD ejection fraction 20+/-6%) and in 31 age-matched controls (29 men). RESULTS: MSNA was higher in HF patients than in controls (51.5+/-14.3 bursts/min versus 33.0+/-11.1 bursts/min; P<0.0001). The VE/VCO(2) slope was also higher in HF patients than in controls (33.7+/-5.7 versus 26.0+/-3.5; P<0.0001), whereas pVO(2) was lower in HF patients than in controls (18.6+/-6.6 versus 31.4+/-8.4 mL/kg/min; P<0.0001). There were significant relationships between MSNA and VE/VCO(2) in both HF (r=0.50; P=0.005) and control subjects (r=0.36; P=0.046). The slope of this regression equation was steeper in HF (0.20 versus 0.11 x MSNA; P=0.001). An analysis of covariance for main effects, including age and pVO(2), identified a significant independent relationship between MSNA burst frequency and VE/VCO(2) (P=0.013) that differed between HF and controls (P<0.01). CONCLUSIONS: The magnitude of resting sympathetic activity correlates positively with the VE/VCO(2) slope. Augmentation of this relationship in HF patients is consistent with the concept that enhanced mechanoreceptor reflex activity exaggerates their ventilatory response to exercise. PMID- 18401468 TI - Mid-term outcomes of off-pump versus on-pump coronary artery bypass graft surgery. AB - OBJECTIVE: To evaluate survival and readmissions to hospital for cardiac events or coronary revascularization (REVASC) in patients having off-pump (OPCAB) versus conventional on-pump (CCAB) coronary artery bypass graft surgery (CABG). METHODS: Of 11,368 consecutive patients undergoing isolated CABG between 1996 and 2002, 514 had OPCAB surgery. Using propensity scores, 503 CCAB patients were randomly matched to 503 OPCAB patients. RESULTS: There were no clinical or statistical differences between the two groups for any prognostic variable. However, OPCAB patients received significantly fewer distal anastomoses than the CCAB group (2.6+/-1.0 versus 3.1+/-1.0; P<0.001). There was no difference in operative mortality (OPCAB 1.0%, CCAB 1.4%; P=0.6), but the OPCAB group had significantly fewer operative strokes (0.2% versus 1.8%; P=0.01). Follow-up was 99.7% complete at 2.2+/-1.2 years (range 0 to 6 years). Twice as many OPCAB patients (n=24) required REVASC compared with the CCAB (n=11) group. The following five-year actuarial outcomes are presented for CCAB and OPCAB, respectively: survival: 77+/ 6%, 76+/-8%, P=0.8; freedom from REVASC: 95+/-3%, 92+/-2%, P=0.02; and cardiac event-free survival: 76+/-5%, 62+/-8%; P=0.05. Cox regression revealed that OPCAB was a significant independent predictor of poorer freedom from REVASC (RR 2.2, 95% CI 1.0 to 4.6; P=0.04) and cardiac event-free survival (RR 1.6, 95%CI 1.1 to 2.2; P=0.02). CONCLUSIONS: The use of OPCAB remains controversial. These results, from this early experience, suggest that despite improved hospital outcomes, the lesser degree of REVASC raises concerns about the need for repeat revascularization in the OPCAB group. PMID- 18401469 TI - Silent ischemia: silent after all? AB - OBJECTIVE: To examine the association of nonpain symptoms in men and women with exercise-related silent ischemia, as well as the independence of these findings from other clinical factors. METHODS: A prospective study of 482 women and 425 men (mean age 58 years) undergoing exercise stress testing with myocardial perfusion imaging. Analyses were performed on 60 women and 155 men with no angina but medical perfusion imaging evidence of ischemia during exercise. MEASURES: The presence of various non-pain-related symptoms. Ischemia is indicated by myocardial perfusion defects on exercise stress testing with single photon emission computed tomography. RESULTS: Women reported more nonangina symptoms than men (P<0.05). They experienced fatigue, hot flushes, tense muscles, shortness of breath and headaches more frequently (P<0.05). Symptoms relating to muscle tension and diaphoresis were associated with ischemia after controlling for pertinent clinical covariates. However, the direction of association differed according to sex and history of coronary artery disease events or procedures. Sensitivity of the detection models showed modest improvements with the addition of these symptoms. CONCLUSIONS: While patients who experience silent ischemia experience a number of nonpain symptoms, those symptoms may not be sufficiently specific to ischemia, nor sensitive in detecting ischemia, to be of particular help to physicians in the absence of other clinical information. PMID- 18401470 TI - Clinical and public health assessment of benefits and risks of statins in primary prevention of coronary events: resolved and unresolved issues. AB - Peer-reviewed, evidence-based recommendations for statin use in primary prevention of cardiovascular events are limited. A narrative review of published randomized controlled trials and meta-analyses was conducted to critically appraise the benefits and risks of statins in primary prevention. Statins effectively reduce plasma concentrations of low-density lipoprotein cholesterol, and reduce the risk of cardiovascular events and death. The greatest benefits are observed in high-risk subjects, such as patients with diabetes or hypertension. Serious cardiovascular events should not be included among serious adverse events because they are efficacy outcomes and are dependent on the baseline risk of patients. Rates of specific serious adverse events, such as cancer and rhabdomyolysis, seem to be similar between the statin and control arms of the clinical trials examined. Thus, the benefits of statins in primary prevention outweigh the risks, particularly among high-risk patients. However, the benefit risk ratio would likely be optimized through interventions designed to increase persistence and adherence in a real-life setting. PMID- 18401471 TI - Statins and primary prevention: is all the evidence in? PMID- 18401472 TI - Pulmonary venous structural remodeling in a canine model of chronic atrial dilation due to mitral regurgitation. AB - OBJECTIVE: Structural remodelling plays an important role in the genesis and maintenance of atrial fibrillation (AF). Although some studies that associate structural remodelling with atrial dilation have been reported, structural pulmonary venous (PV) remodelling due to chronic atrial dilation remains unclear. METHODS: Six sham dogs and five mitral regurgitation (MR) dogs (three months after partial mitral valve avulsion) were studied. Separate cryosections from the PV and left atrium (LA) were immunolabelled with antibodies against connexin (Cx) 40 and Cx43 and analyzed by confocal laser scanning microscopy. Tissue samples from the PV and LA were stained with hematoxylin and eosin, and Masson's trichrome. RESULTS: In MR models, a decrease in Cx40 (0.57+/-0.2% versus 1.18+/ 0.3%, P<0.05) and Cx43 (0.48+/-0.2% versus 1.56+/-0.5%, P<0.05) expression was observed compared with sham dogs. The distribution pattern of Cx40 and Cx43 changed from homogeneous to heterogeneous. Gap junction remodelling was not observed in the LA. In Masson's trichrome-stained sections from MR dogs, regions with increased interstitial fibrosis were present in the PV. Thickness in the PV and the PV-LA junction did not change in the MR group. CONCLUSION: The present study demonstrated a decrease in Cx40 and Cx43 expression and increased interstitial fibrosis in PV due to MR. These changes may potentially be a mechanism that renders the dilated atria more susceptible to AF. PMID- 18401473 TI - Catheter-based local antiproliferative therapy in kissing balloon technique for in-stent stenosis of coronary artery bifurcation lesions. AB - There is currently no promising interventional solution for in-stent stenosis in previously stented bifurcation lesions, even with drug-eluting stents. Rather than being restricted to stent struts, catheter-based local antiproliferative therapy offers the advantage of homogenous drug transfer to the whole vessel wall, and thereby allows for intracoronary pharmacotherapy without adding additional layers of metal into an already stented lesion. The newly developed GENIE catheter (Acrostak Corp, Switzerland), applied in the kissing balloon technique, allows for delivery of liquid paclitaxel into whole bifurcation lesions without repeat stent implantation. After conventional percutaneous transluminal coronary angioplasty, local delivery of paclitaxel using two GENIE catheters in the kissing balloon technique was performed in three patients (left anterior descending, left circumflex and right coronary arteries) with highly symptomatic in-stent bifurcation stenoses. The intervention was feasible and safe in all coronary arteries. Final angiography and control angiography after six months showed good results. No major adverse cardiac events occurred 30 days and six months after intervention. The patients, who represent a group at high risk of recurrent instent restenosis, remained asymptomatic since the local drug delivery. They did not require insertion of a drug-eluting stent or crossover to coronary artery bypass surgery. In conclusion, this new treatment strategy proved to be safe and effective in this first human experience and offers a promising alternative to surgery or implantation of additional stents in these patients. PMID- 18401474 TI - Giant right atrial thrombi treated with thrombolysis. AB - The present report describes giant atrial thrombi that were treated with thrombolysis in a community hospital. Two patients with giant atrial thrombi whose treatment involved complications are presented. Both patients developed cardiogenic shock and were treated unsuccessfully with thrombolysis. Because thrombolysis of giant thrombi may be ineffective, patients in this situation may require surgery. PMID- 18401475 TI - The effectiveness of renal transplantation as a treatment for recurrent uremic cardiomyopathy. AB - Congestive heart failure is more prevalent in patients with end-stage renal disease than in the general population. Although optimal treatment has yet to be defined, these patients are considered to be at high risk for renal transplantation. The present report of a 27-year-old man describes a full recovery from uremic cardiomyopathy following a renal transplant. Despite the patient developing recurrent cardiac dysfunction following graft failure, a second transplant was successful, which, again, resulted in complete resolution of the cardiomyopathy. Patients with recurrent uremic cardiomyopathy following failed kidney transplantation should be considered for a second kidney transplant. PMID- 18401476 TI - Osborn waves in a hypothermic patient with head trauma. PMID- 18401477 TI - Mitral stenosis and cardiogenic shock from an obstructive abscess of the mitral annulus. AB - A 48-year-old female hemodialysis patient with gram-positive mitral endocarditis developed progressive hemodynamic collapse. Echocardiography revealed a mitral annular abscess, which expanded rapidly due to connection with and pressure from the left ventricle. The mitral annular abscess had enlarged sufficiently to obstruct the mitral orifice, resulting in acute cardiogenic shock. The patient underwent and survived emergency surgical repair, and remained well on follow-up. PMID- 18401478 TI - Acute stent thrombosis in a patient with giant cell arteritis. AB - A 70-year-old man presented with a non-ST elevation myocardial infarction attributable to a 99% occlusion of his mid-left anterior descending artery and concomitant symptoms suggestive of giant cell arteritis. He underwent bare metal stenting, with an excellent angiographic result, and was placed on dual antiplatelet therapy, as well as oral prednisone 30 mg twice daily, while awaiting confirmatory temporal artery biopsy. Unfortunately, 70 h after percutaneous coronary intervention, he developed acute stent thrombosis. Platelet aggregometry demonstrated that he was responsive to acetylsalicylic acid, but marginally responsive to clopidogrel. Over the next 30 days, he had a marked clinical improvement, with an improvement in platelet response to clopidogrel that paralleled the clinical resolution of his vasculitis. The present case reports the first incidence of acute stent thrombosis associated with giant cell arteritis, and suggests that the proinflammatory milieu of acute inflammatory arthritides may warrant prolonged, aggressive antiplatelet therapy in the setting of an acute coronary syndrome. PMID- 18401479 TI - Quadricuspid aortic valve in a patient with mitral valve prolapse. PMID- 18401480 TI - Continuing medical education-not enough to simply ''show up''. PMID- 18401481 TI - Orthopedic board certification and physician performance: an analysis of medical malpractice, hospital disciplinary action, and state medical board disciplinary action rates. AB - Specialty board certification status has become the de facto standard of competency by which the profession and the public recognize physician specialists. However, the relationship between orthopedic board certification and physician performance has not been established. Rates of medical malpractice claims, hospital disciplinary actions, and state medical board disciplinary actions were compared between 1309 board-certified (BC) and 154 non-board certified (NBC) orthopedic surgeons in 3 states. There was no significant difference between BC and NBC surgeons in medical malpractice claim proportions (BC, 19.1% NBC, 16.9% P = .586) or in hospital disciplinary action proportions (BC, 0.9% NBC, 0.8% P = 1.000). There was a significantly higher proportion of state medical board disciplinary action for NBC surgeons (BC, 7.6% NBC, 13.0% P = .028). An association between board certification status and physician performance is necessary to validate its status as the de facto standard of competency. In this study, BC surgeons had lower rates of state medical board disciplinary action. PMID- 18401482 TI - Management of late posttraumatic kyphosis with anterior Z-plate instrumentation. AB - Failed treatment of thoracolumbar spine fractures may lead to late posttraumatic kyphosis (LPK), and LPK treatment is challenging. The aim of this retrospective study was to investigate whether anterior reduction and Z-plate instrumentation constitute feasible treatment for LPK (> 30 days after injury). Twenty patients who developed LPK after a thoracolumbar fracture were treated with the Z-plate anterior thoracolumbar plating system. Sixteen patients were followed for a mean of 35 months. Ten of 15 patients with a thoracotomy had persistent postthoracotomy pain. Mean back pain decreased significantly, from 9.2 before surgery to 4.1 after surgery (10 = worst pain ever experienced, 0 = no pain). Osseous union occurred in all patients. Postoperative loss of reduction of 4.9 degrees kyphotic angle was recorded at follow-up. Anterior stabilization with the Z-plate is a technically feasible procedure in patients with LPK. Long-term postthoracotomy pain seems to be a significant problem in these patients. PMID- 18401483 TI - Focal spontaneous osteonecrosis and medial meniscus tear: two cases and a literature review. PMID- 18401484 TI - Transient osteoporosis of the hip in association with osteogenesis imperfecta: two cases, one complicated by a femoral neck fracture. PMID- 18401485 TI - Bilateral tibial tubercle avulsion fractures associated with Osgood-Schlatter's disease. PMID- 18401486 TI - Surgical reconstruction of a late-presenting volar radiocarpal dislocation: a case report. PMID- 18401487 TI - Two cases of missed Salter-Harris III coronal plane fracture of the lateral femoral condyle. AB - Coronal plane fractures of the lateral femoral condyle can be difficult to diagnose, especially in children with open physis. Two adolescents who sustained this uncommon Salter-Harris III fracture of the knee were misdiagnosed after initial clinical examination and standard x-rays. Oblique x-rays, computed tomography, and magnetic resonance imaging were valuable in arriving at the correct diagnosis and in decision making. PMID- 18401488 TI - Using the quick couple drill attachment as an intraoperative metal file. AB - The occasional need to alter or modify plate length during surgery is familiar to most orthopedic surgeons. However, at the modification site, sharp edges may remain. Files or rasps are often used to smooth these edges to prevent local tissue irritation and injury to surgical personnel. To reduce the potential for personal injury during preparation and implantation, while adding convenience and speed to this procedure, we introduce a technique for filing down sharp edges of plates and implants with equipment readily available within most orthopedic operating rooms. PMID- 18401489 TI - Re-examining the safety issues of ceramic-on-ceramic bearing surface. PMID- 18401490 TI - Metallosis after metal-on-polyethylene total hip arthroplasty. AB - Metal debris should not be generated in a well-fixed, well-functioning metal-on polyethylene total hip arthroplasty. However, surgeons sometimes encounter periprosthetic metallosis during revision hip surgery. Insert wear, fracture, or dislodgment in modular components may lead to articulation of the prosthetic head with the metallic shell and subsequent metallosis. Metallosis may occur with loose acetabular components as a consequence of fretting of the screws and shell screw holes or shedding of the ingrowth surface of the component. The femoral component can also be a source of metallosis: Wear of a titanium femoral head, loosening of rough surface finish from the femoral stem, and stem fracture all may result in metallic particles being deposited in periarticular tissues. Specific clinical and radiographic findings can help in differentiating these forms of failure and in planning surgery. When metallic debris-induced bone loss is recognized early, surgical intervention may limit its progression. PMID- 18401491 TI - Ceramic total hip arthroplasty in the United States: safety and risk issues revisited. AB - The advantages of all-alumina bearings are superb wear resistance, stability, and inertness demonstrated over 3 decades. The disadvantage is a small risk for brittle fracture, as described in this paper. Surveying the latest ceramic hip series reported in recent journal articles or presented at the 6th World Biomaterials Congress, we found 11 studies representing more than 35,000 cases followed for 3 to 25 years. There were 24 reported fractures. A unique survey of hip complications in the 1990s found a fracture risk of approximately 1.4 per 1000 ceramic balls used in the United States. A company database holding more than 2.5 million records described the overall fracture risk as 1 per 10,000 cases. Initial use of ceramic cup inserts indicated a 2% to 3% incidence of chipping during surgery. Beginning in 1997, the number of ceramic-metal cup locking cases entered into a US Food and Drug Administration ceramics database was more than 2400, with no fractures reported by the FDA in July 2003. PMID- 18401492 TI - In vitro corrosion analysis in low-intensity, pulsed ultrasound. AB - Clinical investigations have demonstrated a reduced time to union in certain fractures whose management is augmented with low-intensity ultrasound. It is hypothesized that ultrasound augmentation is attributable to mechanical stimulations at the cellular level. Additionally, mechanical stimulation of various magnitudes affects the corrosion rate of metals. Therefore, the effect of ultrasound on the corrosion properties of orthopedic implant materials warrants evaluation prior to recommending ultrasound as an adjunctive treatment for fractures in the presence of internal fixation devices. The purpose of this study was to determine whether low-intensity ultrasound adversely affects the corrosion properties of 316L stainless steel, a commonly used metal in surgical implants. An electrochemical cell was used to expose 316L stainless steel specimens to a corrosion environment. Experimental specimens were subjected to low-intensity ultrasound at the clinically applied intensity. Polarization curves were used to extract average corrosion current density in the passive region, primary passive potentials, and transpassive potentials. Analysis revealed no significant differences between the experimental and control corrosion current density, primary passive potentials, or transpassive potentials. Based on this in vitro analysis, we demonstrated no significant difference in corrosion rate between controls and exposed samples. We conclude that low-intensity ultrasound has no adverse effect on the corrosion properties of stainless steel implant materials. PMID- 18401493 TI - Knee mass from severe metallosis after failure of a metal-backed patellar component total knee arthroplasty. PMID- 18401494 TI - Failed first metatarsophalangeal arthroplasty salvaged by hamstring interposition arthroplasty: metallic debris from grommets. PMID- 18401495 TI - Functionalization of polyoxometalates: towards advanced applications in catalysis and materials science. AB - Functionalization via covalent grafting of organic functions allows to tune the redox and acid-base properties, and the solubility of polyoxometalates, to enhance their stability and biological activity and to reduce their toxicity, to facilitate their implementation in extended structures and functional devices. We discuss herein the electronic and binding connections, and the various synthesis methodologies. We emphasize on organonitrogen, organosilyl and organophosphonyl derivatives with special attention to synthesis, characterization and potential applications in catalysis and materials science. We also consider the giant molybdenum oxide-based clusters especially the porous capsule-type clusters (Keplerates) which have high relevance to this context. PMID- 18401496 TI - Iodide-catalysed self-assembly of donor-acceptor [3]catenanes. AB - Charged donor-acceptor [3]catenanes comprising the pi-accepting cyclobis(paraquat 4,4'-biphenylene) and pi-donating aromatic crown ether macrocycles have been prepared in high yields using thermodynamically controlled dynamic nucleophilic substitution. PMID- 18401497 TI - Rhodamine B thiolactone: a simple chemosensor for Hg2+ in aqueous media. AB - Rhodamine B thiolactone is developed as a simple chemosensor towards Hg(2+) in neutral aqueous solution with high selectivity. PMID- 18401498 TI - Rhodamine thiospirolactone. Highly selective and sensitive reversible sensing of Hg(II). AB - A novel rhodamine thiospirolactone chemosensor was found to develop prominent absorbance and fluorescence enhancements in the presence of Hg(2+) in aqueous solution and this was suggested to result from the thiospiro ring opening induced by Hg(2+) binding. PMID- 18401499 TI - Chemoenzymatic synthesis of prodigiosin analogues--exploring the substrate specificity of PigC. AB - Analogues of prodigiosin, a tripyrrolic pigment produced by Serratia species with potent immunosuppressive and anticancer activities, have been produced by feeding synthetic analogues of the normal precursor MBC to mutants of Serratia sp. ATCC 39006 or to engineered strains of Escherichia coli; in this way it has been shown that the prodigiosin synthesising enzyme, PigC, has a relaxed substrate specificity. PMID- 18401501 TI - A dinuclear ruthenium(II) complex that functions as a label-free colorimetric sensor for DNA. AB - A dinuclear ruthenium(II) complex groove binds to DNA and this interaction results in distinctive color changes that are dependent on both DNA sequence and structure. PMID- 18401500 TI - Role and substrate specificity of the Streptomyces coelicolor RedH enzyme in undecylprodiginine biosynthesis. AB - The function of RedH from Streptomyces coelicolor as an enzyme that catalyses the condensation of 4-methoxy-2,2'-bipyrrole-5-carboxaldehyde (MBC) and 2 undecylpyrrole to form the natural product undecylprodiginine has been experimentally proven, and the substrate specificity of RedH has been probed in vivo by examining its ability to condense chemically-synthesised MBC analogues with 2-undecylpyrrole to afford undecylprodiginine analogues. PMID- 18401502 TI - Size matters--strong binding of the terephthalate dianion by thiourea functionalised fused [n]polynorbornane hosts. AB - Remarkably strong binding of the new [5]polynorbornane based host to the terephthalate dianion is based on size complementarity of the preorganised binding cleft with the rigid dicarboxylate guest. PMID- 18401503 TI - High-level QM/MM modelling predicts an arginine as the acid in the condensation reaction catalysed by citrate synthase. AB - High-level ab initio quantum mechanical/molecular mechanical (QM/MM) modelling of citryl-CoA formation in citrate synthase reveals that an arginine residue acts as the proton donor; this proposed new mechanism helps to explain how chemical and large scale conformational changes are coupled in this paradigmatic enzyme. PMID- 18401504 TI - TNT adsorption on Au(111): electrochemistry and adlayer structure. AB - Electrochemistry and adlayer structure of trinitrotoluene (TNT) on an Au(111) electrode were investigated using cyclic voltammetry and in situ electrochemical scanning tunneling microscopy (ECSTM). PMID- 18401505 TI - Unimolecular binary half-adders with orthogonal chemical inputs. AB - Unimolecular half-adders based upon an arylvinyl-bipyridyl fluorophore platform were demonstrated where all the chemical input combinations were fully processed by half-adder molecules to generate the arithmetic results of the entire truth table. PMID- 18401506 TI - Single microbead SELEX for efficient ssDNA aptamer generation against botulinum neurotoxin. AB - An efficient and easy-to-execute single microbead SELEX approach is developed to generate high affinity ssDNA aptamers against botulinum neurotoxin. PMID- 18401507 TI - Fast energy transfer within a self-assembled cyclic porphyrin tetramer. AB - The structure of a cyclic self-assembled tetramer of an asymmetric meso ethynylpyridyl-functionalized Zn(II)-porphyrin was established by solution-phase X-ray scattering and diffraction; femtosecond transient absorption and anisotropy spectroscopies were used to (a) observe rapid energy transfer (3.8 ps(-1)) between porphyrin subunits and (b) establish that the transfer occurs between adjacent units. PMID- 18401508 TI - Isolation and X-ray structural characterization of tetraisopropylpyrene cation radical. AB - A practical synthesis of 1,3,6,8-tetraisopropylpyrene and the isolation and X-ray structural characterization of its monomeric cation radical salt are described. PMID- 18401509 TI - Self-assembly of a peptide rod-coil: a polyproline rod and a cell-penetrating peptide Tat coil. AB - Peptide rod-coil molecules, composed of a stiff polyproline rod and a hydrophilic cell-penetrating peptide Tat coil, self-assemble into nanocapsules and mediate efficient intracellular delivery of entrapped hydrophilic molecules. PMID- 18401510 TI - A near-infrared dye for multichannel imaging. AB - A large Stokes shift dye, composed of water-solubility and near-infrared feature, was developed for multichannel imaging applications. PMID- 18401511 TI - Unconventional thermodynamically stable cis isomer and trans to cis thermal isomerization in reversibly photoresponsive [0.0](3,3')-azobenzenophane. AB - Sterically hindered [0.0](3,3')-azobenzenophane exhibits thermal trans-to-cis isomerization to the thermodynamically stable cis-cis isomer, and reversible photochemical isomerization with good fatigue resistance. PMID- 18401512 TI - Optical sensing of sulfate by polymethinium salt receptors: colorimetric sensor for heparin. AB - Polymethinium salts based on substituted malondialdehyde have been prepared; the salt with PEG substitution showed high selectivity for sulfate anions and heparin in aqueous medium at physiological condition; intracellular imaging of heparin rich subcellular compartments was achieved with our polymethinium novel receptor for cancer cell lines. PMID- 18401513 TI - Time-dependent growth of zinc hydroxide nanostrands and their crystal structure. AB - Positively-charged crystalline zinc hydroxide nanostrands with a diameter of 2 nm and a length of a few micrometres rapidly grew in dilute aqueous solution of zinc nitrate and aminoethanol. The nanostrands were composed of hexagonal clusters of [Zn(61)(OH)(116)(H(2)O)(n)](6+). PMID- 18401514 TI - A chiral pinwheel supramolecular network driven by the assembly of PTCDI and melamine. AB - The mixing of perylene-3,4,9,10-tetracarboxylic diimide (PTCDI) and 1,3,5 triazine-2,4,6-triamine (melamine) at room temperature in a ratio of 3 : 4 on Au(111) leads to the formation of a new chiral "pinwheel" structure. PMID- 18401515 TI - Ni-nitrilotriacetic acid-modified quantum dots as a site-specific labeling agent of histidine-tagged proteins in live cells. AB - Ni-nitrilotriacetic acid (NTA) functionalized CdSe/ZnS quantum dots (QDs) were exploited as a site-specific labeling agent of histidine-tagged biomolecules in live cells; the QDs were found to be water-soluble, aggregation free and stable for several months. PMID- 18401516 TI - Fluorescence microscopy coupled to electrochemistry: a powerful tool for the controlled electrochemical switch of fluorescent molecules. AB - The first-time coupling of fluorescence microscopy with a three-electrode electrochemical cell is described and applied to the investigation of a controlled-potential redox switch of organic fluorophores such as tetrazine derivatives. PMID- 18401517 TI - Preparation of silyl substituted crotylzinc reagents and their highly diastereoselective addition to carbonyl compounds. AB - Readily prepared beta-silyl substituted crotylzinc reagents undergo highly selective allylation of carbonyl compounds leading to syn-homoallylic alcohols. PMID- 18401518 TI - Synthesis and reactivity of tetrakis(imino)pyracene (TIP) ligands; bifunctional analogues of the BIAN ligand class. AB - The first two examples of a new class of bifunctional BIAN-type ligand have been prepared, and the reactions of one such ligand with CuBr(2) and BCl(3) have been explored. PMID- 18401519 TI - One-pot synthesis of reverse type-I In2O3@In2S3 core-shell nanoparticles. AB - A novel method to one-pot-synthesize high-quality In(2)O(3)@In(2)S(3) core-shell nanoparticles, consisting of a step of reducing In(2)O(3) core surface into a layer of active indium metal in high-temperature organic solution and a step of converting this layer to In(2)S(3) using CS(2), has been developed. PMID- 18401520 TI - A new NCN pincer ruthenium complex and its catalytic activity for enantioselective hydrogenation of ketones. AB - New bis(oxazolinyl)phenyl-ruthenium(II) complexes, which were synthesized by C-H bond activation with RuCl(3).3H(2)O in zinc powder and 1,5-cyclooctadine followed by ligand exchange reaction with sodium acetylacetonate or acetylacetone, exhibited enantioselective hydrogenation of ketones in up to 90% ee. PMID- 18401521 TI - An unusual dianion equivalent from acylsilanes for the synthesis of substituted beta-keto esters. AB - The reaction of lithiated diazo esters with acylsilanes generates a remarkable intermediate en route to highly substituted beta-keto esters. PMID- 18401522 TI - Novel fluorene-based functional 'click polymers' for quasi-solid-state dye sensitized solar cells. AB - Quasi-solid-state DSSCs in SnO(2):F/quasi-solid-state electrolyte/Pt devices have been fabricated with click polymers for the first time; they show a maximum power conversion efficiency (PCE) of 2.80%. PMID- 18401523 TI - Phase-transfer catalytic aza-Michael addition of tert-butyl benzyloxycarbamate to electron-deficient olefins. AB - A highly efficient phase-transfer catalytic aza-Michael addition of tert-butyl benzyloxycarbamate to a wide range of electron-deficient olefins is presented (90 99%). PMID- 18401524 TI - Activity assay of membrane transport proteins. AB - Membrane transport proteins are integral membrane proteins and considered as potential drug targets. Activity assay of transport proteins is essential for developing drugs to target these proteins. Major issues related to activity assessment of transport proteins include availability of transporters, transport activity of transporters, and interactions between ligands and transporters. Researchers need to consider the physiological status of proteins (bound in lipid membranes or purified), availability and specificity of substrates, and the purpose of the activity assay (screening, identifying, or comparing substrates and inhibitors) before choosing appropriate assay strategies and techniques. Transport proteins bound in vesicular membranes can be assayed for transporting substrate across membranes by means of uptake assay or entrance counterflow assay. Alternatively, transport proteins can be assayed for interactions with ligands by using techniques such as isothermal titration calorimetry, nuclear magnetic resonance spectroscopy, or surface plasmon resonance. Other methods and techniques such as fluorometry, scintillation proximity assay, electrophysiological assay, or stopped-flow assay could also be used for activity assay of transport proteins. In this paper the major strategies and techniques for activity assessment of membrane transport proteins are reviewed. PMID- 18401525 TI - Inhibitor of apoptosis proteins and apoptosis. AB - Apoptosis is a physiological cell death process that plays a critical role in development, homeostasis, and immune defense of multicellular animals. Inhibitor of apoptosis proteins (IAPs) constitute a family of proteins that possess between one and three baculovirus IAP repeats. Some of them also have a really interesting new gene finger domain, and can prevent cell death by binding and inhibiting active caspases, but are regulated by IAP antagonists. Some evidence also indicates that IAP can modulate the cell cycle and signal transduction. The three main factors, IAPs, IAP antagonists, and caspases, are involved in regulating the progress of apoptosis in many species. Many studies and assumptions have been focused on the anfractuous interactions between these three main factors to explore their real functional model in order to develop potential anticancer drugs. In this review, we describe the classification, molecular structures, and properties of IAPs and discuss the mechanisms of apoptosis. We also discuss the promising significance of clinical applications of IAPs in the diagnosis and treatment of malignancy. PMID- 18401526 TI - Zinc finger protein 278, a potential oncogene in human colorectal cancer. AB - Zinc finger protein 278 (ZNF278) is a novel Krueppel Cys2-His2-type zinc finger protein that is ubiquitously distributed in human tissues. Whether ZNF278 is related to the development of colorectal cancer is still unclear. The transcriptional level of ZNF278 was studied in colorectal cancer by real-time polymerase chain reaction. The results showed that ZNF278 expression was increased in 53% of colorectal cancer tissues compared to corresponding non cancerous tissues. The transcriptional down-regulation of ZNF278 was detected in only three (6%) human colorectal cancer tissues compared to corresponding non cancer tissues. No significant difference was detected in 19 (41%) pairs of samples. However, we failed to find a significant association between the up regulation of ZNF278 transcription and age, sex, the degree of infiltration, or the tumor size of colorectal cancer. To study the function of ZNF278 in colorectal carcinogenesis, the colon cancer cell line SW1116 was stably transfected with a wild-type ZNF278 plasmid to construct an overexpression system, and was transiently transfected with the small interfering RNA of ZNF278 to construct a ZNF278 knockdown system. Cell proliferation was assessed with 3 [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide dye and a cell counter. The results show that ZNF278 promotes cell growth, and its knockdown suppresses cell proliferation. ZNF278 could be a potential proto-oncogene in colorectal cancer. PMID- 18401527 TI - Inhibition of fibroblast growth factor 2-induced apoptosis involves survivin expression, protein kinase C alpha activation and subcellular translocation of Smac in human small cell lung cancer cells. AB - To investigate the mechanism by which fibroblast growth factor 2 (FGF-2) inhibits apoptosis in the human small cell lung cancer cell line H446 subjected to serum starvation, apoptosis was evaluated by flow cytometry, Hoechst 33258 staining, caspase-3 activity, and DNA fragmentation. Survivin expression induced by FGF-2 and protein kinase C alpha (PKC alpha) translocation was detected by subcellular fractionation and Western blot analysis. In addition, FGF-2-induced release of Smac from mitochondria to the cytoplasm was analyzed by Western blotting and immunofluorescence. FGF-2 reduced apoptosis induced by serum starvation and up regulated survivin expression in H446 cells in a dose-dependent and time dependent manner, and inhibited caspase-3 activity. FGF-2 also inhibited the release of Smac from mitochondria to the cytoplasm induced by serum starvation and increased PKC alpha translocation from the cytoplasm to the cell membrane. In addition, PKC inhibitor inhibited the expression of survivin. FGF-2 up-regulates the expression of survivin protein in H446 cells and blocks the release of Smac from mitochondria to the cytoplasm. PKC alpha regulated FGF-2-induced survivin expression. Thus, survivin, Smac, and PKC alpha might play important roles in the inhibition of apoptosis by FGF-2 in human small cell lung cancer cells. PMID- 18401528 TI - Angiotensin II suppresses adriamycin-induced apoptosis through activation of phosphatidylinositol 3-kinase/Akt signaling in human breast cancer cells. AB - Angiotensin II (Ang II) stimulates tumor growth and angiogenesis in some solid cancer cells, but its anti-apoptosis role in breast cancer remains unclear. To address this issue, we investigated the effect of Ang II on adriamycin-induced apoptosis in breast cancer MCF-7 cells. Treatment of human breast cancer MCF-7 cells with adriamycin, a DNA topoisomerase II alpha inhibitor, caused apoptosis. However, cells pretreated with Ang II were resistant to this apoptosis. Ang II significantly reduced the ratio of apoptotic cells and stimulation of phospho-Akt Thr308 and phospho-Akt-Ser473 in a dose-dependent and time-dependent manner. In addition, Ang II significantly prevented apoptosis through inhibiting the cleavage of procaspase-9, a major downstream effector of Akt. The Ang II type 1 receptor (AT1R) was responsible for these effects. Among the signaling molecules downstream of AT1R, we revealed that the phosphatidylinositol 3-kinase/Akt pathway plays a predominant role in the anti-apoptotic effect of Ang II. Our data indicated that Ang II plays a critical anti-apoptotic role in breast cancer cells by a mechanism involving AT1R/phosphatidylinositol 3-kinase/Akt activation and the subsequent suppression of caspase-9 activation. PMID- 18401529 TI - Apelin-13 induces ERK1/2 but not p38 MAPK activation through coupling of the human apelin receptor to the Gi2 pathway. AB - Apelin signaling to the family of mitogen-activated protein kinases (MAPKs), such as extracellular-regulated kinases 1/2 (ERK1/2) and p38 MAPK, through the coupling of apelin receptor (APJ) to G-protein, mediates important pathophysiological responses. Although apelin fragments have been reported to induce ERK1/2 activation through Gi-protein, the intracellular pathways by which APJ activates these MAPKs are only partially understood. Here, using stably transfected human embryonic kidney 293 (HEK293) cells overexpressing human APJ (HEK293-apelinR), we showed that apelin-13 signaling leads to ERK1/2 and p38 MAPK pathways through APJ activation. It was found in HEK293-apelinR cells that ERK1/2 activation was initiated by apelin-13 at 5 min, with the peak of activation occurring at 15 min, and a return to the basal level within 60 min. The activation of ERK1/2 appeared to be dose-dependent with a significant activation being observed at 10 nM apelin-13 and maximal activation at 100 nM. However, phosphorylated-p38 MAPK was not detected in HEK293-apelinR cells treated with apelin-13. We also shown that the apelin-13-induced ERK1/2 activation requires a coupling with pertussis toxin-sensitive G-protein, and that overexpression of dominant-negative Gi2 completely inhibits the apelin-13-induced ERK1/2 activation. In addition, treatment with apelin-13 resulted in a concentration dependent reduction of forskolin-stimulated cAMP production. It is therefore suggested that apelin-13 activates ERK1/2 but not p38 MAPK, which involves the coupling of APJ to the Gi2 cascade. In conclusion, the ERK1/2, but not p38 MAPK pathway is activated by apelin-13 through coupling of human APJ to Gi2-protein, which contributes to cellular responses. PMID- 18401530 TI - Alternative method for site-directed mutagenesis of complex polyketide synthase in Streptomyces albus JA3453. AB - Sequence analysis of oxazolomycin (OZM) biosynthetic gene cluster from Streptomyces albus JA3453 revealed a gene, ozmH, encoding a hybrid polyketide and non-ribosomal peptide enzyme. Tandem ketosynthase (KS) domains (KS 10-1 and KS 10 2) were characterized and they show significant homology with known KSs. Using an alternative method that involves RecA-mediated homologous recombination, the negative selection marker sacB gene, and temperature-sensitive replications, site directed mutagenesis of the catalytic triad amino acid cysteines were carried out in each of the tandem KS domains to test the function they play in OZM biosynthesis. HPLC-mass spectrometry analysis of the resulting mutant strains showed that KS 10-2 is essential for OZM biosynthesis but KS 10-1 is not indispensable and might serve as a "redundant" domain. These results confirmed the existence of an "extra domain" in complex polyketide synthase. PMID- 18401531 TI - Immunization against Egyptian Schistosoma mansoni infection by multivalent DNA vaccine. AB - The development of multivalent vaccines consisting of several antigens is a novel approach to creating broad-range protection against different parasite strains and parasite life cycle stages. We have previously confirmed that the schistosome Sm21.7 and SmFimbrin (SmFim) proteins could induce protection in mice. Therefore, this study aimed to construct the multivalent DNA vaccine Sm21.7-SmFim/pBudCE4.1 and evaluate its immune efficacy. The open reading frames of two Schistosoma mansoni genes, Sm21.7 and SmFim, were inserted into the eukaryotic expression plasmid pBudCE4.1 designed for the independent expression of two genes in mammalian cells. To evaluate the in vitro expression of the multivalent Sm21.7 SmFim/pBudCE4.1 DNA vaccine and its immunological effect in mice, the recombinant plasmid Sm21.7-SmFim/pBudCE4.1 was used to transfect 293T cells, and the expression of mRNA and proteins was examined using reverse transcription polymerase chain reaction and Western blot analysis. Then the ability of Sm21.7 SmFim/pBudCE4.1 to protect against S. mansoni challenge infections was analyzed according to worm burden and egg reduction rates after vaccination of mice. Vaccinated mice showed a significant level of protection (56%), and a decrease in the number and size, and change in the cellular profile, of granulomas. Egg reduction in liver and intestine was 41.53% and 55.63%, respectively, as determined relative to mice that received the empty vector only. In addition to reductions in worm viability, worm fecundity and egg hatching ability were observed following challenge infection in the immunized group. Results showed that Sm21.7-SmFim/pBudCE4.1 could express Sm21.7 and SmFim mRNA and proteins. Enzyme-linked immunosorbent assay and Western blot analysis indicated that immunized mice generated specific immunoglobulin G against Sm21.7 SmFim/pBudCE4.1. These results suggest that vaccination with multivalent S. mansoni DNA vaccine (SmFim-Sm21.7/pBudCE4.1) not only induces a significant reduction in worm and egg burdens, but also significantly reduces the size of egg granulomas. In summary, the multivalent vaccine stimulated specific immunity with a significant level of protection and has anti-pathological effect. PMID- 18401532 TI - Glucose-regulated protein 75 suppresses apoptosis induced by glucose deprivation in PC12 cells through inhibition of Bax conformational change. AB - Glucose-regulated protein 75 (Grp75) is an important molecular chaperone that belongs to the heat shock protein 70 family and resides predominantly in mitochondria. Grp75 can protect cells from glucose deprivation (GD) injury. However, the molecular mechanisms by which it carries out this function are unknown. Here we report that Grp75 could delay the release of cytochrome c and reduce apoptosis induced by GD, and we also found that Grp75 could decrease Bax/Bcl-2 gene expression ratio and inhibit the conformational change of Bax during this process. In conclusion, these findings suggested that Grp75 overexpression was able to inhibit apoptosis induced by GD. Grp75 inhibited Bax conformational change to delay the release of cytochrome c, thus providing protection to PC12 cell which was used primarily as a neuron model against GD toxicity. PMID- 18401533 TI - High expression of osteoglycin decreases the metastatic capability of mouse hepatocarcinoma Hca-F cells to lymph nodes. AB - Osteoglycin, one of the matrix molecules, belongs to the small leucine-rich proteoglycan gene family and might play important roles in cell growth and differentiation and in pathological processes such as fibrosis and cancer growth. In this study, a eukaryotic expression plasmid pIRESpuro3 osteoglycin(+) was constructed and transfected into mouse hepatocarcinoma Hca-F cells to evaluate the contribution of osteoglycin to the malignant behavior of Hca-F. It was found that Hca-F cells transfected with pIRESpuro3 osteoglycin(+) showed significantly decreased potential for both migration and invasion. Furthermore, Hca-F cells transfected with osteoglycin showed decreased metastatic potential to peripheral lymph nodes. However, proliferation potential and adhesive capacity of Hca-F cells to different protein substrates were not influenced by osteoglycin transfection. In summary, high expression of osteoglycin decreases the metastatic capability of Hca-F to lymph nodes. PMID- 18401534 TI - Cloning, expression, and polymorphism of the porcine calpain10 gene. AB - Calpains are calcium-regulated proteases involved in cellular functions that include muscle proteolysis both ante- and post-mortem. This study was designed to clone the complete coding sequence of the porcine calpain10 gene, CAPN10, to analyze its expression characteristics and to investigate its polymorphism. Two isoforms of the CAPN10 gene, CAPN10A and CAPN10B, were obtained by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends methods combined with in silico cloning. RT-PCR results indicated that CAPN10 mRNA was ubiquitously expressed in all tissues examined and, with increasing age, the expression level increased in muscles at six different growth points. In the same tissues, the expression level of CAPN10A was higher than that of CAPN10B. In addition, three single nucleotide polymorphisms were detected by the PCR-single-stranded conformational polymorphism method and by comparing the sequences of Chinese Min pigs with those of Yorkshire pigs. C527T mutation was a missense mutation and led to transforming Pro into Leu at the 176 th amino acid. The results of the current study provided basic molecular information for further study of the function of the porcine CAPN10 gene. PMID- 18401535 TI - Sorption of acetochlor, atrazine, 2,4-D, chlorotoluron, MCPA, and trifluralin in six soils from Slovakia. AB - Sorption of the herbicides (acetochlor, atrazine, 2,4-D: , chlorotoluron, MCPA and trifluralin) in their commercially available formulations was characterized on six agricultural soils from Slovakia. Weak acid herbicides (2,4-D: and MCPA) were the least sorbed, whereas weak base such as atrazine and nonionic herbicides were the most sorbed in the order: atrazine < acetochlor approximately chlorotoluron < trifluralin, which was closely related to the hydrophobicity of herbicides expressed as logKOW. The KF values positively correlated with soil organic carbon content for all herbicides. Moreover, all herbicides were the most sorbed by Vertisol with relatively high smectite clay content in proportion to organic carbon content. The KOC values for each herbicide showed a smaller variation among soils (excluding Vertisol) than the KF and were used to compare soil mobility potential of herbicides. PMID- 18401536 TI - Giant intravesical calculus during pregnancy. AB - Urolithiasis is commonly found during pregnancy; but the presence of a giant vesical calculus during pregnancy is a very rare entity, associated with several potential obstetric complications. A 25-year-old primigravida at 25 weeks of gestational age was referred to our tertiary care unit because she presented a giant hyperechoic intravesical mass and inability to pass urine with suprapubic pain since 2 days. An open cystolithotomy revealed a huge intravesical calculus. The patient continued with her pregnancy until full term without adverse perinatal outcomes. PMID- 18401537 TI - Serious complication 1 year after sacrospinous ligament fixation. AB - Myositis of the gluteal region caused by group A streptococci 1 year after a sacrospinous ligament fixation was recognised as a serious complication of this procedure. Most likely, the infection was spread to the gluteal region through a port d'entree caused by vaginal atrophy, via the non-resorbable sutures. The patient was treated successfully with antibiotics intravenous and local estrogens. PMID- 18401538 TI - Urinary tract infections after pelvic floor gynecological surgery: prevalence and effect of antimicrobial prophylaxis. A systematic review. AB - We evaluated the prevalence of urinary tract infection (UTI) after pelvic floor operations for non-malignant etiology and the effectiveness of antibiotic prophylaxis. This was made possible by a review of the evidence from relevant randomized controlled trials (RCTs). Nineteen out of 879 initially identified studies met the criteria for inclusion in our review. Four RCTs compared an antibiotic prophylactic regimen with placebo, 11 two different prophylactic antibiotic regimens, and four had three different treatment arms. Among placebo recipients undergoing pelvic floor surgery, 10-64% developed UTI. In contrast, UTI after pelvic floor gynecological surgery occurred in 0-15% of the patients who received cephalosporins as antibiotic prophylaxis; the likelihood for postoperative UTI was higher for patients receiving cotrimoxazole (28%), ampicillin/sulbactam (13.6%), metronidazole plus ampicillin (20%), metronidazole (10-22.7%), or ciprofloxacin (27.2%). The use of a cephalosporin as perioperative antimicrobial prophylaxis is the optimal regimen in preventing UTIs after pelvic floor surgery. PMID- 18401539 TI - Intrathecal chemotherapy delivered by a lumbar-thecal catheter in metastatic medulloblastoma: a case illustration. AB - BACKGROUND: Medulloblastoma is the most common malignant brain tumour in children. Despite recent advances, the prognosis in high risk patients remains poor. Further improvement in survival is dependent upon the development of strategies to attack the tumour more effectively, but with less toxicity. Intrathecal chemotherapy, is an ideal but currently underused method of directly targeting residual tumour within the area of resection and the leptomeningeal disease commonly associated with this tumour. METHOD: We describe the case of a 12 yr old child with metastatic medulloblastoma, who received intrathecal topotecan via a spinal catheter. CONCLUSION: This method represents a simple, safe and effective method of delivering an even and widespread distribution of drug within the cerebrospinal fluid (CSF) of the neuroaxis. With new agents being identified and others in the early stages of development, intrathecal chemotherapy may emerge as an important therapeutic option to consider when faced with such challenging cases. PMID- 18401541 TI - Using ensemble of classifiers for predicting HIV protease cleavage sites in proteins. AB - The focus of this work is the use of ensembles of classifiers for predicting HIV protease cleavage sites in proteins. Due to the complex relationships in the biological data, several recent works show that often ensembles of learning algorithms outperform stand-alone methods. We show that the fusion of approaches based on different encoding models can be useful for improving the performance of this classification problem. In particular, in this work four different feature encodings for peptides are described and tested. An extensive evaluation on a large dataset according to a blind testing protocol is reported which demonstrates how different feature extraction methods and classifiers can be combined for obtaining a robust and reliable system. The comparison with other stand-alone approaches allows quantifying the performance improvement obtained by the ensembles proposed in this work. PMID- 18401540 TI - Proteasomal abnormalities in cortical Lewy body disease and the impact of proteasomal inhibition within cortical and cholinergic systems. AB - Dementia with Lewy bodies (DLB) accounts for 15-20% of the millions of people worldwide with dementia. In the current work we investigate the association between proteasome dysfunction and the development of cortical Lewy body pathology. Analysis of post-mortem cortical tissue indicated levels of the alpha subunit of the 20S proteasome were significantly reduced in DLB cortex, but not Alzheimer's, in comparison to control and this reduction correlated with both the severity and duration of dementia. Application of proteasome inhibitors to rodent cortical primary neurones in vitro and by direct injection onto rodent cholinergic forebrain neurons in vivo gave rise to dose dependent neuronal death and in rodent cortex -- marked cholinergic deficits accompanied by the accumulation of inclusions that stained positive for alpha-synuclein and ubiquitin. These findings suggest that proteasomal abnormalities are present within cortical Lewy body disease and the experimental inhibition of proteasomal function mirrors the neuropathological changes seen within the disorder. PMID- 18401542 TI - Human Delta1-pyrroline-5-carboxylate synthase: function and regulation. AB - Mammalian Delta(1)-pyrroline-5-carboxylate synthase (P5CS) is a bifunctional ATP- and NAD(P)H-dependent mitochondrial enzyme that catalyzes the coupled phosphorylation and reduction-conversion of L: -glutamate to P5C, a pivotal step in the biosynthesis of L: -proline, L: -ornithine and L: -arginine. Previously, we reported cloning and characterization of two P5CS transcript variants generated by exon sliding that encode two protein isoforms differing only by a two amino acid-insert at the N-terminus of the gamma-glutamyl kinase active site. The short form (P5CS.short) is highly expressed in the gut and is inhibited by ornithine. In contrast, the long form (P5CS.long) is expressed ubiquitously and is insensitive to ornithine. Interestingly, we found that all the established human cell lines we have studied expressed P5CS.long but not P5CS.short. In addition, expression of P5CS.long can be modulated by hormones: downregulation by hydrocortisone and dexamethasone and upregulation by estradiol, for example. Using a quantitative proteomic approach, we showed that P5CS.long is upregulated by p53 in p53-induced apoptosis in DLD-1 colorectal cancer cells. Functional genomic analysis confirmed that there are two p53-binding consensus sequences in the promoter region and in the intron 1 of the human P5CS gene. Interestingly, overexpression of P5CS by adenoviruses harboring P5CS.long or P5CS.short in various cell types has no effect on cell growth or survival. It would be of importance to further investigate the role of P5CS as a p53 downstream effector and how P5CS.short expression is regulated by hormones and factors of alternative splicing in cells isolated from model animals. PMID- 18401543 TI - The metabolism of proline, a stress substrate, modulates carcinogenic pathways. AB - The resurgence of interest in tumor metabolism has led investigators to emphasize the metabolism of proline as a "stress substrate" and to suggest this pathway as a potential anti-tumor target. Proline oxidase, a.k.a. proline dehydrogenase (POX/PRODH), catalyzes the first step in proline degradation and uses proline to generate ATP for survival or reactive oxygen species for programmed cell death. POX/PRODH is induced by p53 under genotoxic stress and initiates apoptosis by both mitochondrial and death receptor pathways. Furthermore, POX/PRODH is induced by PPARgamma and its pharmacologic ligands, the thiazolidinediones. The anti tumor effects of PPARgamma may be critically dependent on POX/PRODH. In addition, it is upregulated by nutrient stress through the mTOR pathway to maintain ATP levels. We propose that proline is made available as a stress substrate by the degradation of collagen in the microenvironmental extracellular matrix by matrix metalloproteinases. In a manner analogous to autophagy, this proline-dependent process for bioenergetics from collagen in extracellular matrix can be designated "ecophagy". PMID- 18401544 TI - Victimization and bullying among 8-year-old Finnish children: a 10-year comparison of rates. AB - OBJECTIVE: There are only few population-based time-trend studies on changes in prevalence of bullying and victimization among children. The main aim of this study was to find out whether changes have occurred in prevalence rates of bullying and victimization from 1989 to 1999 among eight-year-old children. The associations between victimization and bullying and psychiatric symptoms and their possible differences in the statistical strengths of associations between the years 1989 and 1999 were also studied. METHOD: Two cross-sectional, representative samples from southern Finland were compared. All children born in 1981 (1989 sample, n = 985, response rate 95%) and 1991 (1999 sample, n = 962, response rate 86%) and living in the selected school district were included in the study samples. Children, parents and teachers were asked about bullying and victimization. The Children's Depression Inventory and Rutter's parent and teacher scales were used to study psychiatric symptoms. RESULTS: In 1999, fewer 8 year-old children were victims of bullying than in 1989. There was a decrease in the number of bullies but the change was statistically significant only in the parental reports. The statistical strengths of associations of victimization and bullying with psychiatric symptoms were mainly the same in 1989 as in 1999. CONCLUSIONS: Slightly decreased levels of victimization among 8-year-old Finnish children is a promising result, but further time-trend studies are needed, as well as qualitative studies, to obtain a deeper understanding of the bullying phenomenon and the factors reducing it. It might be that especially young children are suspectible to influences diminishing victimisation and bullying. PMID- 18401545 TI - A rare case of primary actinomycosis of the anterior abdominal wall: diagnosis and treatment. AB - We present the case of a 60-year-old man who presented with a left hypochondrial swelling first noticed 4 weeks prior to admission to our clinic. Based on the findings of the ultrasound and magnetic resonance imaging investigation, a tumour of uncertain origin of the abdominal wall was suspected, also involving the small bowel. The swelling, including the affected lateral and transverse oblique muscles as well as the subcutaneous tissue and the adjacent omentum majus, was completely excised. The resulting myoaponeurotic defect of the left lateral abdominal wall was closed with interrupted Vicryl sutures and stabilised with a PTFE prosthesis (20 x 10 cm) that was placed intraabdominally and secured by spiral tackers and interrupted transfascial monofilament Prolene sutures. Microscopic examination of the excised specimen revealed an actinomycosis of the anterior abdominal wall, which is extremely rare. The surgical treatment was followed by antibiotic therapy for 6 months. This treatment resulted in full recovery with no further complications. PMID- 18401546 TI - Positive association of the FTSJ1 gene polymorphisms with nonsyndromic X-linked mental retardation in young Chinese male subjects. AB - To investigate the possible genetic association of nonsyndromic X-linked mental retardation (NS-XLMR) with FTSJ1 gene polymorphisms, a case-control association study was performed focusing on the Chinese Han population in the Qinba mountain region. Three common single nucleotide polymorphisms (SNPs) (rs2268954, rs2070991, rs5905692) in the gene were selected and genotyped using the polymerase chain reaction single-strand confirmation polymorphism (PCR-SSCP) method. Pairwise linkage disequilibrium (LD) analysis showed that the three SNPs were in strong LD (all D' > 0.8). There were significant differences between cases and controls in allele frequency distribution of rs2268954 (P = 0.036), rs2070991 (P = 0.043), and rs5905692 (P = 0.014) and in the distributions of common haplotypes combined by these SNPs (global P = 0.01236) in male subjects. In female subjects, however, no positive results were found. Our results suggest a positive association between the genetic variants of the FTSJ1 gene and NS-XLMR in young male subjects in the Chinese Han population in the Qinba region. PMID- 18401547 TI - TIM-3 polymorphisms in type 1 diabetes families. AB - TIM-3 is a transmembrane protein preferentially expressed on differentiated Th1 cells, which play a role in Th1-mediated diseases including type 1 diabetes. We investigated the role of the rs10515746 (A/C), rs1036199 (A/C), and rs10053538 (A/C) single nucleotide polymorphisms (SNPs) within the TIM-3 gene in 186 German type 1 diabetes families (558 individuals) and its interaction with human leukocyte antigen (HLA) high-risk haplotypes DQ2(DQA*0501-DQB*0201)-DQ8 (DQA*0301 DQB*0302). Alleles A, C, and A of the rs10515746 (A/C), rs1036199 (A/C), and rs10053538 (A/C) SNPs were found in a frequency of 20.4%, 19.0%, and 4.2%, respectively. Transmission analysis of these polymorphisms did not show any significant difference. Although in patients with HLA DQx/x (neither HLA DQ2 nor DQ8) an undertransmission of allele A (14.3% vs. 85.7%) of the rs10053538 (A/C) SNP and an overtransmission of allele A (66.7% vs. 33.3%) of the rs10515746 (A/C) SNP was observed, these associations did not remain statistically significant after correction for multiple comparisons. Although we found no association of TIM-3 with type 1 diabetes in the German population, we cannot discard a possible association in a larger size. PMID- 18401548 TI - Formation of downhill esophageal varices as a rare but serious complication of hemodialysis access: a case report and comprehensive literature review. AB - BACKGROUND: Proximal or "downhill" esophageal varices are a rare complication of superior vena caval (SVC) obstruction. Few reports describe downhill varices in dialysis patients with catheter-related SVC occlusion. METHODS: We studied a case of downhill esophageal varices in a dialysis patient from our center and reviewed the published literature on presentation, evaluation and treatment in other dialysis patients (MEDLINE database search). RESULTS: Including our current case, we identified eight reports of dialysis patients with downhill varices. All cases were recognized after presentation with gastrointestinal bleeding, in contrast to low reported bleeding rates of downhill varices in non-dialysis patients. Localized edema and superficial venous engorgement (signs of SVC occlusion) were each observed in four of eight patients. The duration of hemodialysis dependence ranged from 2.5 to 23 years, and dialysis access history included multiple central venous catheters when described (seven cases). Central venous imaging by direct, magnetic resonance or computerized tomographic venography documented SVC stenosis in all cases. Management included percutaneous transluminal angioplasty of the SVC with or without stenting in five of eight patients, three of whom developed restenosis during observation. Successful surgical venous bypass was performed in one patient after failed percutaneous venoplasty. Varices were treated with band ligation in four of eight cases without reported complications. CONCLUSIONS: Although rare, downhill esophageal varices should be considered in the differential diagnosis of upper gastrointestinal hemorrhage in dialysis patients exposed to central venous catheters. Diagnosis should prompt radiographic evaluation of SVC patency. Treatment requires timely and coordinated care by specialists in endovascular interventions and gastrointestinal endoscopy. PMID- 18401550 TI - Autoenucleation in a 84-year-old dementia patient. AB - BACKGROUND: Autoenucleation is a severe, rare form of self-mutilation. The majority of cases have been reported in the 15- to 60-year age group, usually in psychiatric patients with a history of depression or schizophrenia, sometimes caused by drug abuse. CASE REPORT: We report a case of left-sided autoenucleation in an 84-year-old dementia patient suffering from reactive depression. Medical reports mentioned a suicide attempt 2 weeks prior to the incident, whereupon the patient was admitted to the locked ward of a psychiatric hospital. During one night of inpatient stay, the patient manually autoenucleated his left eye. Inspection of the enucleated organ revealed a completely intact globe with an attached optic nerve 5.5 cm in length. The orbit was filled with a massive haematoma. A contrast-enhanced computed tomography (CT) scan showed an orbital haematoma, absence of the left globe and optic nerve and a chiasmatic lesion, accompanied by an intracranial bleeding into the subarachnoid space. Primary wound closure was performed without complications. Visual acuity of the right eye could not be tested due to the patient's lack of cooperation. CONCLUSION: To the best of our knowledge, this is the only reported case of an elderly patient with primary dementia who performed autoenucleation. Other aspects, such as patient history, suicide attempt, manual eye extraction and chiasma lesion are similar to cases reported earlier. The identification and evaluation of intracranial bleedings and chiasmatic lesions that can be associated with autoenucleation requires a contrast-enhanced CT, especially if a long optic nerve fragment is attached to the enucleated globe. PMID- 18401549 TI - Intracranial hemodynamics during intravenous infusion of glyceryl trinitrate. AB - The mechanisms of glyceryl trinitrate (GTN)-induced headache are not fully elucidated. In this study we administered GTN 0.5 microg/kg/min i.v. for 20 min in six healthy volunteers. Before, during and 60 min after the infusion, we investigated regional cerebral blood flow (rCBF), cerebral blood volume (CBV), both estimated with SPECT, and blood flow velocity (BFV) in the middle cerebral artery (MCA), measured with transcranial Doppler. Headache was scored on a numerical verbal rating (0-10) scale. rCBF was unchanged, CBV was slightly increased (13%) during GTN infusion, whereas BFV decreased both during (20%) and 60 min (15%) after GTN. Headache was short-lived and maximal during infusion. This discrepancy of time-effect curves for the effect of GTN on headache and dilatation of MCA indicates that MCA is most likely not the primary source of pain in GTN-induced headache. The time-effect curves for the effect of GTN on headache and on dilation of MCA differed markedly. This indicates that MCA is most likely not the primary source of pain in GTN-induced headache. PMID- 18401551 TI - Effective temporary analgesia for severe painful blind eye. AB - There are various methods of providing pain relief for painful blind eyes. We wish to recommend this effective method of providing temporary analgesia in patients suffering from a severe painful blind eye before undergoing enucleation. PMID- 18401552 TI - Intravitreal bevacizumab (Avastin) as primary and rescue treatment for choroidal neovascularization secondary to ocular toxoplasmosis. AB - BACKGROUND: Treatment modalities for choroidal neovascularization due to ocular toxoplasmosis include laser photocoagulation, surgery, corticosteroids, and verteporfin therapy. Intravitreal injection of bevacizumab in the treatment of choroidal neovascularization due to other conditions appears to be an effective and safe therapeutic option. CASE REPORTS: We report two young patients with choroidal neovascularization secondary to ocular toxoplasmosis who received a single intravitreal injection of bevacizumab as primary or rescue therapy. After a follow-up of 12 and 10 months, respectively, visual acuity improved, and features of active neovascularization resolved with no recurrence. No adverse events were recorded. CONCLUSION: Intravitreal injection of bevacizumab appears to be an effective and safe treatment modality in patients with choroidal neovascularization secondary to ocular toxoplasmosis. Further evaluation with a longer follow-up period is needed to confirm these findings. PMID- 18401553 TI - Implantation of scleral fixated sutured posterior chamber lenses: a retrospective analysis of 119 cases. AB - BACKGROUND: The implantation of scleral fixated sutured posterior chamber lenses enables lens implantation in the absence of adequate lens capsule support. Anterior chamber lenses and their side effects can also be avoided, particularly in cases of unilateral aphakia and contact lens incompatibility. METHODS: This study is a retrospective analysis of 119 scleral fixated sutured posterior chamber lenses implanted in 115 patients. Seventy-nine cases of damaged zonula or damaged lens capsule and 17 cases of corneal decompensation secondary to anterior chamber lenses were the main indications for the suture fixation. The mean follow up was 23 months (6-83 months). RESULTS: In 50% of all cases, improved visual acuity was achieved. Twenty-four percent of patients showed unchanged visual acuity, while 26% had poorer vision postoperatively, mainly because of corneal decompensation following anterior chamber lens explantation. The main postoperative complication was transient rise of intraocular pressure, occurring in 44% of the cases. CONCLUSIONS: In most cases improved or comparable postoperative visual acuity was achieved. The main complication observed was postoperative glaucoma, especially in patients with pre-existing glaucoma. PMID- 18401554 TI - Isolated foveal hypoplasia. AB - PURPOSE: To describe a patient with isolated foveal hypoplasia. METHODS: A 55 year-old man with the clinical suspicion of foveal hypoplasia was given a complete ophthalmological examination, including optical coherence tomography (OCT), fluorescein angiography (FA) and fundus-related perimetry (FRP). Mutation screening for oculocutaneous albinism and aniridia was also performed, but the results were negative for both. RESULTS: Following a complete ophthalmological examination and genetic studies, we were able to confirm the clinical suspicion of isolated foveal hypoplasia in this otherwise healthy patient. CONCLUSIONS: With this report we want to highlight the roles of OCT, FA and FRP in the diagnosis of such a singular condition as isolated foveal hypoplasia. PMID- 18401555 TI - Visual outcome in cataract surgery complicated by vitreous loss in a district general hospital. AB - PURPOSE: To determine the visual outcome of patients following cataract surgery complicated by vitreous loss and to establish the factors, if any, that influenced the final visual outcome. METHODS: Retrospective review of all patients over a 1-year period who had planned phacoemulsification complicated by vitreous loss. Information was entered into a pre-devised proforma and the results analysed. RESULTS: Of the 1614 eyes operated on for cataract, 43 (2.66%) had surgery complicated by vitreous loss. The mean age of the patients experiencing surgical complications was 79.7 years (range 48-93 years). Twenty three patients (53%) had at least one factor contributing to the difficulty of procedure, with dense cataract being the top factor in 11 patients. There was no statistically significant association between any pre-operative predisposing factor, stage at which vitreous loss occurred, grade of the surgeon and final visual outcome. When pre-existing co-morbidity was excluded, 26 of 30 (87%) patients achieved a best-corrected visual acuity of 6/12 or better post operatively. Cystoid macula oedema was the underlying cause in 50% of those patients that lost one or more lines post-operatively. CONCLUSIONS: Surgeons at all levels of training had an acceptable complication rate, with the majority of patients achieving a good visual outcome. Dense cataract was the most common factor contributing to the complexity of the surgery. PMID- 18401556 TI - Endothelial nitric oxide synthase gene haplotypes and diabetic nephropathy among Asian Indians. AB - Endothelial dysfunction plays a key role in the pathogenesis of diabetic vascular disease, including diabetic nephropathy. Endothelial-derived nitric oxide synthase (eNOS) gene polymorphisms affect eNOS activity and are associated with endothelial dysfunction. We evaluated the association of the constitutive endothelial nitric oxide synthase gene (eNOS) polymorphisms with type 2 diabetic nephropathy. We genotyped three polymorphisms of eNOS (Two SNPs: -786T > C, 894G > T and one 27-bp repeat polymorphism in Intron 4 (27VNTR)) in type 2 diabetic nephropathy patients (cases: n = 195) and type 2 diabetic without nephropathy (controls: n = 255), using validated PCR-RFLP assays. We measured serum NO levels in these subjects and examined its correlation with diabetic nephropathy and eNOS genotypes. The frequency of CC (-786T > C), TT (894G > T) and aa genotypes (27VNTR) were significantly higher in diabetic nephropathy patients as compared to the diabetes without nephropathy group (CC: P = 0.003, TT: P = 0.03, aa: P < 0.0001). These mutant genotypes were found to be associated with higher risk of nephropathy (-786T > C: OR: 5.5, 95%CI: 1.53-19.79; 894G > T: OR: 1.8, 95%CI: 1.03-3.16; Intron 4: OR: 6.23, 95%CI: 2.23-16.31). Haplotype with all the wild alleles (T-b-G) was found to be associated with a decreased risk of nephropathy (OR: 0.68, P = 0.005) and haplotype with all mutant alleles (C-a-T) was associated with higher risk of diabetic nephropathy as compared to diabetes without nephropathy group (OR: 2.6, P = 0.14). No significant linkage disequilibria were observed among the variants in this case-control study. The serum NO levels were observed to be significantly (P < 0.05) lower in mutant allele carriers ('C' allele of T-786C SNP and/or 'T' allele of G894T SNP) as compared with the wild-type allele carriers (-786T and/or 894G) within each of the subject groups (with and without nephropathy). These results suggest that the eNOS gene locus is associated with diabetic nephropathy and the functional polymorphisms (-786T > C & 894G > T) might lead to a decreased expression of eNOS gene. PMID- 18401557 TI - Contribution of the conservative cleavage motif to posttranslational processing of the carboxyl terminal domain of rodent Muc3. AB - In previous works, we showed by transient expression studies in COS-1 cells that the C-terminal domain of rat intestinal membrane mucin (rMuc3) that was cloned in the pSecTag2 plasmid (named as p20) is posttranslationally cleaved twice. One location is between the glycine and the serine within a LS1KGS2IV1V2 motif, and the other is in the 49 kDa membrane-tethered fragment at an undefined site. The sea-urchin sperm protein, enterokinase and agrin module of rMuc3 is responsible for the cleavage and association of the cleaved fragments. The present study demonstrates how the conservative cleavage motif LS1KGS2IV1V2 contributes to posttranslational processing through mutagenesis of each residue in the LS(1)KGS2IV1V2 motif. Mutation of S2 to alanine (p20s2/a) completely prevented cleavage. While p20k/a (in this construct the K is replaced by A) and p20s1/a (in this construct the S1 is replaced by A) (6 and 3%) showed almost the same result as the wild-type p20 transfectant (4%), 79, 39, 22, 17, and 14% of the products from p20g/a (in this construct the G is replaced by A), p20i/a (in this construct the I is replaced by A), p20l/a (in this construct the L is replaced by A), p20v2/a (in this construct the V2 is replaced by A), and p20v1/a (in this construct the V1 is replaced by A) remained uncleaved. The cleaved N-terminal fragment of the p20s1/a transfectant was 26 kDa, but the N-terminal fragments from p20, p20g/a, p20l/a, p20k/a, p20i/a, p20v1/a, and p20v2/a were 30 kDa. The S1 residue was possibly O-glycosylated, which was supported by deglycosylation with O-cocktail (a mixture of glycosidases). The N-terminal fragment of p20s1/a transfected cells was present at high levels in the spent media. Thus, the S2, G, I, L, V2, and V1 residues within the conserved cleavage motif, LS1KGS2IV1V2, are important for cleavage and contribute to the structural formation and conformational stress of the small loop between the beta2 and the beta3 strands. The S1 residue is possibly O-glycosylated, and mutation of S1 residue to alanine does not affect the cleavage of the LS1KGS2IV1V2 motif, but it is important for the dissociation and further release of the cleaved N-terminal fragment from the cell surface. PMID- 18401558 TI - Influence of gender and oral contraceptives intake on innate and inflammatory response. Role of neuroendocrine factors. AB - The purpose of the present work was to determine differences between young men (M), and women in the follicular phase (W) and women taking oral contraceptives containing ethinylestradiol (CW) in the phagocytic process of neutrophils (chemotaxis, phagocytosis and microbicide capacity), in the serum concentrations of cytokines both pro-inflammatory (IFNgamma, TNFalpha, IL-12, IL-6, IL-8) and anti-inflammatory ones (IL-10 and IL-13), and in neuroendocrine factors with immunomodulatory capacity (estradiol, prolactin, cortisol, catecolamines and 72 kDa heat shock proteins, Hsp72). Men showed a lower phagocytosis and microbicide capacity than women, and less serum concentrations of the pro-inflammatory cytokines IL-6 and IL-8. CW neutrophils also showed a lower phagocytic capacity than W neutrophils, together with less serum IL-8 concentration. CW showed the highest serum concentration of IL-13. However, no statistical changes were observed in the pro-inflammatory cytokines: INF-gamma, TNF-alpha, IL-12 and in the anti-inflammatory cytokine IL-10. The greater anti-inflammatory status in CW than in W was parallel with lower concentrations of oestrogens. Cortisol, prolactin, and the extracellular Hsp72 seem to be involved in the gender- and contraceptives-induced differences in the inflammatory response. While cortisol (in general an immunosuppressive hormone) showed the highest values in CW, prolactin and Hsp72 (an immunopermissive factors) showed the lowest values in CW and M. Less clear is the participation of catecholamines in the gender inflammatory differences. PMID- 18401560 TI - New method showing the influence of matrix components in Leuconostoc mesenteroides biofilm formation. AB - Studying biofilm formation and influence of the matrix composition was heavy because only old and long methods were employed up to now: confocal microscopy, fluorescent chemical markers, and/or dying techniques. In this context, an innovative tool, the BioFilm Ring Test, was here employed to explore the role of exopolysaccharides, proteins, and nucleic acids in the formation of biofilm by Leuconostoc mesenteroides. The principle is to add magnetic particles in the culture medium. When a biofilm is formed, particles are unable to migrate in the media to form a ring when a magnet is brought nearer to the well. Therefore, culture media supplemented with proteases, glycanases, and/or nucleases allowed us to identify the involvement of these substances in L. mesenteroides biofilm formation. The results permitted to demonstrate that dextran, proteins, and nucleic acids are implied in biofilm formation. PMID- 18401559 TI - Efficacy of DL-alpha lipoic acid against systemic inflammation-induced mice: antioxidant defense system. AB - Inflammation can activate macrophages or monocytes and sequentially release several inflammatory cytokines and reactive oxygen species (ROS). Oxidative stress-induced acute inflammatory response plays an important role in several diseases. This study was designed to investigate the prophylactic effect of the antioxidant lipoic acid (LA) during inflammation-induced mice. Mice were divided in to three groups (n = 8 in each): control, systemic inflammation, and LA treated mice with systemic inflammation. Results show that ROS was significantly higher in lymphocytes, hepatocytes, and astrocytes (P < 0.05) of inflammation induced mice when compared with control but no significant changes were observed in the LA treated group. Increased levels of lipid peroxidation (LPO) and decreased activities of oxidants such as superoxide dismutase (SOD), catalase, glutathione peroxidase, glutathione, and ATPase were observed in the inflammation induced mice, which returned to near normalcy following LA therapy. In vitro study has shown that LA treatment not only suppresses the increased LPO levels but also inhibits the lipid break down resulting from autoxidation. In addition, increased immunoreactivity of the astrocyte marker glial fibrillary acidic protein (GFAP) was observed in the neocortex region of inflammation-induced mice, whereas nuclear factor kappa B p65 (NFkappaB) immunoreactivity was observed in both the neocortex and liver of the same group which were effectively controlled by LA therapy suggesting that LA can efficiently manage systemic inflammation. PMID- 18401561 TI - Detection of virulence genes in Vibrio cholerae isolated from aquatic environment in Kerala, Southern India. AB - Vibrio cholerae is the etiologic agent of cholera. It is an autochthonous inhabitant of all aquatic environments. The virulence of V. cholerae is maintained by the CTX genetic element and tcpA gene. In the present investigation, environmental strains of V. cholerae isolated from different aquatic biotopes in Kerala were identified and serotyped. The antibiotic resistance pattern and presence of virulence and regulatory genes were examined. We found the presence of toxigenic non-O1/non-O139 strains harboring the CTX genetic element, heat-stable enterotoxin, rtxA gene, El Tor hemolysin, and Vibrio pathogenicity island (VPI). The strains also produced the cholera toxin (CT) as determined by monosialoganglioside enzyme-linked immunosorbent assay. A few strains belonging to the O1 serogroup but lacking the CTX genetic element were also observed. The majority of the environmental strains belonged to non-O1/non O139 serogroup with many possessing toxR, ompU, heat-stable enterotoxin, and rtxA gene. The toxigenic non-O1/non-O139 strains exhibited resistance to trimethoprim, ampicillin, and polymixin B and intermediate resistance to co-trimoxazole. However, all other environmental strains were found resistant to ampicillin and polymixin B. Our findings demonstrate that the virulence genes are dispersed among the environmental strains of V. cholerae and a complex aquatic environment can give rise to pathogenic V. cholerae. PMID- 18401562 TI - Specific entities affecting the craniocervical region: osteogenesis imperfecta and related osteochondrodysplasias: medical and surgical management of basilar impression. AB - INTRODUCTION: Osteogenesis imperfecta (OI) is an inheritable disorder of bone development caused by defective collagen synthesis. The attendant basilar impression or secondary basilar invagination is uncommon but can be devastating. CLINICAL MATERIALS AND METHODS: Fifty-two patients with osteochondrodysplasia (28 with OI, six with Hajdu-Cheney syndrome, six with Paget's disease, and 12 with spondyloepiphyseal dysplasia) with basilar impression were evaluated between 1985 and 2005. The male/female ratio in this cohort was 1:1. The mean age at presentation was 12.2 years. SYMPTOMS: Symptoms and signs included headache, lower cranial nerve dysfunction, dysphagia, respiratory embarrassment, weakness, and ataxia. TREATMENT: In the earlier part of the series (1985-1995), all patients with hydrocephalus were shunted and a ventral transoral decompression made for ventral compression of the pontomedullary junction followed by a dorsal occipitocervical fusion. As a result of this evaluation, it was felt that most patients would benefit by early bracing after the hydrocephalus was shunted if it existed. However, 20% of patients still required an anterior ventral decompression and the occipitocervical fusion. RESULTS: The results showed that the fusions were stable but over a period of time, there was progressive forward bending with osteogenesis imperfecta as well as with the Hajdu-Cheney syndrome. All patients with spondyloepiphyseal dysplasia had a good strong stable fusion which stood the test of time. CONCLUSION: In conclusion, we feel that early intervention with occipitocervical bracing can prevent the progressive march of significant basilar impression which leads to mortality. PMID- 18401563 TI - Craniocervical developmental anatomy and its implications. AB - INTRODUCTION: Congenital and developmental osseous abnormalities and anomalies that affect the craniocervical junction complex can result in neural compression and vascular compromise and can manifest itself with abnormal cerebrospinal fluid dynamics. An understanding of the development of the craniocervical junction is essential to recognize the pathological abnormalities. MATERIALS AND METHODS: Atlas assimilation, segmentation failures, os odontoideum, basilar invagination, and the various syndromes that affect the craniocervical junction have been analyzed. The natural history provides an added insight into its treatment. RESULTS: Proatlas segmentation abnormalities surrounded the foramen magnum and the posterior arch of C1. Hindbrain herniation was associated in 33 of the 90 children involved. Spastic quadriparesis presented in 80% and lower cranial nerve abnormalities in 33%. Vertebrobasilar dysfunction was observed in 40% and trauma presentation seen in 60% of individuals. Atlas assimilation was present in 550 individuals who were evaluated for craniovertebral junction abnormalities. Hindbrain herniation occurred in 38%. Segmentation failure of C2 and C3 vertebrae compounded the abnormal dynamics resulting in atlantoaxial instability. This was a reducible instability with formation of pannus around the odontoid process until it became irreducible at approximately 14 years of age. Unilateral atlas assimilation caused torticollis in children. Os odontoideum was investigated regarding craniocervical trauma at a young age. CONCLUSION: The conclusion was that os odontoideum was associated with an unrecognized fracture in children below the age of 5 with a previously normal odontoid structure as observed in our series. Atlas and axis abnormalities were reviewed in this series. This large database has provided an understanding of the natural history of many entities and allowed treatment protocols to be established that have stood the test of time. PMID- 18401564 TI - Craniovertebral junction neoplasms in the pediatric population. AB - INTRODUCTION: The incidence of tumors at the craniovertebral junction in the pediatric population is low. Because of the variable pathology and the rarity of these tumors, ideal therapies are only now being defined. MATERIALS AND METHODS: Thirty-eight children with tumors affecting the craniocervical junction were encountered between 1991 and 2006. These comprised neoplasms of osseous origin and neural extramedullary tumors. RESULTS AND DISCUSSION: Chordomas of the clivus and foramen magnum were seen in eight, fibrous dysplasia in four, aneurysmal bone cysts in four, eosinophilic granuloma affecting the atlas and axis vertebra in four, Ewing's sarcoma involving the atlas in two, osteoblastoma in two, neurenteric cysts in four, meningioma in five, schwannoma in two, and plexiform neurofibromas in three. The location of these tumors was predominantly ventral, and a very small number had a lateral or dorsal location. The ventral tumors included chordoma, meningioma, fibrous dysplasia, aneurysmal bone cyst, and osteoblastoma. Plexiform neurofibroma affecting the craniocervical junction was ventral to the clivus and upper cervical spine causing severe kyphosis of the craniocervical region. Pain in the head and neck occurred in 70%. Paresthesias and dysesthesias in the hands were seen in 40% and spastic weakness of extremities in 22%. Cranial nerve palsies were seen in 33%. Twenty-eight percent of children showed dysphagia or dysarthria. The cranial nerves affected were the vagus followed by hypoglossal and glossopharyngeal nerves. This led to dysphagia, slurred speech, repeated aspiration pneumonia, and weight loss. The most common findings for chordomas at the craniocervical junction were isolated hypoglossal nerve palsy. All individuals underwent magnetic resonance imaging, computed tomography, and 3D computed tomography and angiography. Vertebral angiography was used to understand the dynamics of collateral circulation and tumor vascularity. Tumor embolization was performed in chordoma and aneurysmal bone cysts. Our experience and results are presented here. PMID- 18401565 TI - Relative toxicity of malathion to trematode-infected and noninfected Rana palustris tadpoles. AB - Amphibian populations around the world are facing threats that include disease and pollution. Although the effect of environmental contaminants on susceptibility to infection has been demonstrated for several amphibian species, to our knowledge, the opposite interaction, infection status affecting contaminant susceptibility, has not been studied. We conducted standard 48-h toxicity tests to compare susceptibility to malathion, a widely used organophosphate insecticide, of uninfected pickerel frog (Rana palustris) tadpoles and tadpoles infected with two levels (10 or 30 cercariae) of the trematode Echinostoma trivolvis. Trematode encystment rates were high (>90%) in both trematode treatment groups. LC(50) values ranged from 16.5 to 17.4 mg/L, within the range reported for other amphibian species. However, we found no differences in susceptibility to malathion among parasite treatments. Although we detected no effect of parasites on pesticide susceptibility in this system, it is important to investigate this question using other pesticides, parasites, and amphibian hosts before dismissing this potentially threatening interaction. PMID- 18401566 TI - Different roles of galectin-9 isoforms in modulating E-selectin expression and adhesion function in LoVo colon carcinoma cells. AB - Galectin-9, a member of galectin family, plays multiple roles in a variety of cellular functions, including cell adhesion, aggregation, and apoptosis. Galectin 9 also has three isoforms (named galectin-9L, galectin-9M, and galectin-9S), but whether these isoforms differ in their functions remains poorly understood. In this study, we showed that transient expression of galectin-9L decreased E selectin levels, while transient expression of galectin-9M or galectin-9S increased E-selectin levels in LoVo cells, which do not express endogenous galectin-9. We also found that over-expression of three galectin-9 isoforms led to increased attachment of LoVo cells to extracellular matrix proteins respectively, while over-expression of galectin-9M or galectin-9S increased the adhesion of LoVo cells to human umbilical vein endothelial cells in vitro. In summary, these findings indicate that different isoforms of galectin-9 exhibit distinct biological functions. PMID- 18401567 TI - C-reactive protein gene polymorphisms affect plasma CRP and homocysteine concentrations in subjects with and without angiographically confirmed coronary artery disease. AB - Human C-reactive protein (CRP) is a reactant involved in the acute phase response and one of the many molecular factors involved in pathogenesis of coronary artery disease (CAD). CRP gene variants potentially mediate CRP plasma concentrations and the development of CAD. 220 Croatian subjects with angiographically confirmed CAD and 132 control subjects were included in the study. CRP gene polymorphisms 1059G/C and -717G/A were determined by RFLPs, using MaeIII and KspI endonuclease, respectively. Plasma concentrations of CRP and homocysteine were determined by immunoturbidimetry and FPIA, respectively. CRP 1059G/C gene variants were significantly associated with CAD (OR = 0.50; 95% CI = 0.27, 0.94; P = 0.032). Wild GG genotype and rare allele C carrier genotypes were 184 and 22 in CAD(+) group, and 101 and 24 in CAD(-) group, respectively. Multivariate analysis with age, gender, BMI, smoking status, hypertension and diabetes as covariates showed that 1059C carriers had lower CRP concentrations in CAD(-) (P = 0.010) and CAD(+) subjects (P = 0.028). This allele was also significantly associated with lower plasma homocysteine concentrations in both groups (P = 0.018 for CAD(-) and 0.002 for CAD(+). There was no significant difference between CAD(+) and CAD(-) subjects in absolute frequencies for CRP -717A/G gene variant, but multivariate analysis showed that carriers of the rarer G allele had significantly higher CRP plasma concentrations in CAD(-) subjects (P = 0.031) and higher homocysteine concentrations in CAD(+) group (P < 0.001). Atherosclerosis is an inflammatory disease resulting from different genetic and environmental factors. Results presented here support the contribution of CRP genetic variations in the development of CAD. PMID- 18401568 TI - Type I and II circumcaval ureter in children: experience in three cases. AB - Circumcaval ureter is a congenital abnormality in which the right ureter passes behind the inferior vena cava (IVC), but it is seldom reported in childhood. Surgery is required for symptomatic cases to transect and reposition the right ureter to lie anterior to the IVC in order to correct this abnormality. We present three childhood cases where two types of obstructive circumcaval ureter were diagnosed and treated surgically. PMID- 18401570 TI - [Replacement therapy with adrenal steroids]. AB - Replacement with adrenal steroids comprises treatment with glucocorticoids, mineralocorticoids and adrenal androgen precursors, mainly in patients with adrenal insufficiency. Attention has also been directed to replacement treatment with glucocorticoids in critically ill patients or with dehydroepiandrosterone (DHEA) in elderly people with an age related decline of DHEA/DHEAS levels. Despite the use of current replacement concepts well-being is often not fully restored in patients with adrenal insufficiency. Innovations comprise the development of new delayed release glucocorticoid preparations that allow to better mimic the circadian cortisol secretion and may have the potential to improve the treatment of patients with adrenal insufficiency. Recent results from the CORTICUS study have not confirmed previous beneficial results and have led to restrictions in the use of hydrocortisone substitution therapy in patients with septic shock. Treatment with the androgen precursor DHEA in patients with adrenal insufficiency may improve quality of life. However, in patients with an age related decline of DHEA levels, the available evidence does not support the use of DHEA replacement as an anti aging treatment. PMID- 18401571 TI - Eggshell colour does not predict measures of maternal investment in eggs of Turdus thrushes. AB - The striking diversity of avian eggshell colour has long fascinated biologists. Recently, it has been proposed that the blue-green colour of some eggs may function as a post-mating sexually selected signal of female phenotypic quality to their mates to induce higher allocation of paternal care. It has been suggested that maternally deposited yolk carotenoids may be the specific aspect of reproductive quality that the female is signalling via eggshell colour. We use the known properties of the thrush visual system (Turdus sp.) to calculate photon capture for the four single cone photoreceptors, and the principal member of the double cone class for eggs in clutches of two introduced European thrush species (Turdus merula and Turdus philomelos) in New Zealand. We show that differences in the avian-perceived colours of individual eggs are not consistently correlated with different measures of maternal investment in the egg. Given the growing extent of the knowledge between maternal quality, parental investment and eggshell pigmentation across avian taxa, we encourage the use of avian perceptual modelling for testing alternative non-signalling explanations for the structural and physiological basis of these relationships. PMID- 18401572 TI - [Acute ischemic stroke. Diagnostic imaging and interventional options]. AB - Stroke is the third most common cause of death after myocardial infarction and neoplasms in industrialized countries and the most common cause for permanent disability with impairment of an independent life style. In addition to the socioeconomic problems caused by a disabling stroke, it is to be expected that with an increasing average age of the population, the number of stroke patients will increase as well [4]. The need for effective and widely available therapies against this severe disease is highly evident. Diagnostic imaging is indispensable in order to apply these therapies efficiently and precisely. In addition to the established intravenous thrombolytic therapy with rt-PA within the first 3 h, a therapeutic benefit can also be achieved with thrombolysis inside the time-window 3-6 h, whereas the rate of symptomatic intracerebral hemorrhages increases. Local intraarterial fibrinolysis (LIF) within 6 h is effective and safe and may lead to considerable improvement despite an initially severe medical condition [13]. Besides LIF, interventional techniques for mechanical recanalization of intracranial vessel occlusions are becoming increasingly more established. According to international guidelines for the treatment of acute stroke, computed tomography (CT) is considered to be the most important technical diagnostic tool if available round-the-clock. Alternatively, magnetic resonance tomography (MRT) may primarily be performed if carried out without delay and if the imaging protocol contains a sequence suitable for exclusion of hemorrhages. PMID- 18401573 TI - [Acute chest pain: a purely clinical problem or a question for radiology?]. AB - Acute chest pain represents a very common clinical occurrence and at the same time poses a severe diagnostic dilemma. It can be due to an acute life threatening event such as acute cardiac infarct, or a relatively harmless condition of pain and illness (e.g. vertebrogenic pain) under the main symptom category of acute chest pain. This often unclear symptomatic, behind which there can always be a life-threatening disease leads to an exaggerated grouping of patients into emergency cases and to an increased number of inpatients for observation. The diagnosis of acute coronary syndrome with no initial ECG changes typical for ischemia is especially problematic. The availability of modern multidetector computed tomography is becoming increasingly more important for radiologists in the diagnosis and clarification of acute chest pain. In this article the clinical difficulties and radiology options for the diagnosis of patients with acute chest pain will be presented and possible future algorithms for diagnosis will be discussed. PMID- 18401574 TI - Boron toxicity in rice (Oryza sativa L.). I. Quantitative trait locus (QTL) analysis of tolerance to boron toxicity. AB - Boron toxicity tolerance of rice plants was studied. Modern japonica subspecies such as Koshihikari, Nipponbare, and Sasanishiki were tolerant, whereas indica subspecies such as Kasalath and IR36 were intolerant to excessive application of boron (B), even though their shoot B contents under B toxicity were not significantly different. Recombinant inbred lines (RILs) of japonica Nekken-1 and indica IR36 were used for quantitative trait locus (QTL) analysis to identify the gene responsible for B toxicity tolerance. A major QTL that could explain 45% of the phenotypic variation was detected in chromosome 4. The QTL was confirmed using a population derived from a recombinant inbred line which is heterogenic at the QTL region. The QTL was also confirmed in other chromosome segment substitution lines (CSSLs). PMID- 18401575 TI - Transient inhibitory effect of methoxychlor on testicular steroidogenesis in rat: an in vivo study. AB - Methoxychlor, an organochlorine pesticide, has been reported to induce reproductive abnormalities in male reproductive tract. To get more insight into the mechanism(s) of gonadal toxicity provoked by methoxychlor, we investigated whether treatment with methoxychlor at low observed adverse effect level (LOAEL) would alter the activities of steroidogenic enzymes such as Delta(5)3beta hydroxysteroid dehydrogenase (3beta-HSD) and Delta(5)17beta-hydroxysteroid dehydrogenase (17beta-HSD), the expression levels of steroidogenic acute regulatory (StAR) protein and androgen binding protein (ABP) in the testis of adult male rats. The experimental rats were exposed to a single dose of methoxychlor (50 mg/kg body weight) orally. The rats were killed at 0, 3, 6, 12, 24 and 72 h following treatment using anesthetic ether and testes were collected, processed and used to measure the activities of 3beta-HSD, 17beta-HSD, levels of hydrogen peroxide produced and the expression levels of StAR protein, and ABP. Methoxychlor administration resulted in a sequential reduction in the expression of StAR protein and activities of 3beta-HSD, 17beta-HSD with concomitant increase in the levels of hydrogen peroxide in the testis. These changes were significant between 6-12 h following treatment. The levels of ABP declined at 6-12 h following exposure to methoxychlor. The present study demonstrates transient effect of methoxychlor at LOAEL on testicular steroidogenesis and the possible role of hydrogen peroxide in mediating these effects. PMID- 18401576 TI - Interdisciplinary study for the evaluation of biochemical alterations on mussel Mytilus galloprovincialis exposed to a tributyltin-polluted area. AB - An interdisciplinary approach was employed to monitor the concentration and the effects of butyltin compounds in mussels (Mytilus galloprovincialis). Tissues from animals exposed to a marine area (Vado Ligure harbour) with a high concentration of tributyltin (TBT) were analysed and compared with control samples. TBT concentrations were measured by gas chromatography-mass spectrometry and the protein pattern in gill tissues was studied by proteomic analysis. Several proteomic signatures associated with contaminant exposure were observed; spots that were significantly increased in all contaminated samples were identified by mass spectrometry as fragments of beta-tubulin. The degradation of beta-tubulin was then confirmed by western blot analysis with specific anti-beta tubulin antibody. The effects observed on mussel gills after exposure in the TBT polluted area are discussed. PMID- 18401577 TI - Safety aspects of iodinated contrast media related to their physicochemical properties: a pharmacoepidemiology study in two Tuscany hospitals. AB - BACKGROUND: More than 380,000 angiographic procedures are performed every year in Italian hospitals, with an increase rate of 8% per year. Although contrast media (CM) are considered relatively safe, adverse drug reactions (ADRs) remain an important issue. OBJECTIVES: The objective of this study was to quantify the incidence of immediate and delayed nonrenal ADRs to iodinated CM in an Italian cohort and to evaluate whether their different physicochemical properties are able to affect the incidence of immediate or delayed ADRs. METHODS: A prospective intensive monitoring study was conducted on a cohort of patients undergoing radiodiagnostic procedures with iodinated CM enrolled in two hospitals in Tuscany, Italy. To evaluate both immediate (within 1 h after CM administration) and delayed (>1 h to 1 week after CM administration) ADRs to CM, two questionnaires were administered. Adverse events (AEs) were analyzed to check the causality assessment between CM and ADR. If more than one symptom occurred in the same patient, they were treated as a single event. RESULTS: One thousand five hundred and fourteen subjects who were exposed to iodinated CM completed the questionnaires. Mean age [standard deviation (SD)] was 65.4 (13.3) years, and 57.9% were male patients. A total of 178 [11.8%; 95% confidence interval (CI) 10.1-13.4] ADRs were reported. Thirty-four (2.2%; 1.5-3.1) and 144 (9.5%; 8.0 11.1) developed immediate and delayed ADRs, respectively. Both types of ADRs were experienced by six subjects (0.4%; 0.1-0.8). One hundred and seventy-six cases (98.8%; 96.0-99.8) were classified as possible and two (1.1%; 0.1-3.9) as probable ADRs. Monomeric low-osmolal (iopromide, iomeprol, iobitridol) and dimeric iso-osmolal (iodixanol) groups mainly reported delayed allergy-like ADRs of mild severity. Only one immediate reaction was severe. Multivariate analysis confirmed a higher risk of immediate reactions occurring for monomeric CM (OR 4.3; 95% CI 1.2-15.7), whereas the risk of delayed ADRs was significantly higher for the dimeric group (OR 1.8; 1.1-2.5). CONCLUSIONS: Monomeric CM were more frequently involved in immediate ADRs, whereas dimeric CM were involved in delayed reactions. Although severe life-threatening ADRs to CM were confirmed to be rare, due to the large use of these drugs, they still retain clinical and epidemiological significance. PMID- 18401578 TI - The effect of paroxetine on the pharmacokinetics, safety, and tolerability of ramosetron in healthy subjects. AB - OBJECTIVE: The aim of our study was to investigate the effects of multiple doses of paroxetine on the pharmacokinetics, safety, and tolerability of a single oral 10-microg dose of ramosetron. METHODS This was an open, one-sequence crossover design study. On day 1, healthy male and female subjects were administered a single dose of 10 microg ramosetron. On the morning of day 3, the subjects were administered paroxetine to reach steady state, which consisted of morning doses of 20 mg on days 3-12. The dose on day 11 was administered in combination with a single dose of 10 microg ramosetron. RESULTS: In subjects genotyped as extensive CYP2D6 metabolizers, coadministration of paroxetine with ramosetron resulted in an increase in area under the curve from 0 to infinity (AUC(0-inf)) and the peak concentration (Cmax) of ramosetron by 1.14-fold (90% confidence interval (CI): 1.07-1.22) and by 1.06-fold (90% CI: 1.00-1.11), respectively. CONCLUSIONS: It can be concluded that the single-dose pharmacokinetic profile of ramosetron 10 microg is not affected to a clinically relevant degree by paroxetine 20 mg once daily administered for 10 days. PMID- 18401579 TI - HIV-1-cellular interactions analyzed by single virus tracing. AB - Single virus tracing (SVT) allows the direct investigation of the entry pathway of viruses into living cells. Using fluorescently labeled virus-like particles (VLPs) and SVT, we have studied the interaction between human immunodeficiency virus type 1 (HIV-1) and the plasma membrane of living cells. From the trajectories of freely diffusing VLPs in solution, we established that the particle preparation was homogeneous and the particles had a hydrodynamic radius of 86 +/- 5 nm, consistent with the size of single HI viruses. The VLPs that come in contact with the cell surface either become immobilized or rapidly dissociate from the cell surface. The fraction of virions that become immobilized on the plasma membrane correlates with the surface heparan sulfate linked proteoglycans (HSPG) concentration of the cell line tested. The particles that are not immobilized make an average of 1.5 contacts with the cell surface before diffusing away. For most cell lines investigated, the contact duration follows an exponential distribution with a lifetime between 20 and 50 ms depending on the cell type. PMID- 18401580 TI - Central skeletal sarcoidosis mimicking metastatic disease. AB - Sarcoidosis is a systemic disease that histologically typically shows non caseating granulomas. The most common radiologic finding is hilar and mediastinal adenopathy. Patients with widely disseminated disease may show involvement of the peripheral appendicular skeleton in 1-13% of such cases. A primary skeletal presentation without other manifestations typical of the disease is rare. We present a case of sarcoidosis in a middle-aged Caucasian man in whom the disease presented with widespread lytic lesions in the axial skeleton and long bones, mimicking metastatic disease. There was no involvement of the peripheral skeleton, skin or lungs. PMID- 18401581 TI - Magnetic resonance imaging of the femoral trochlea: evaluation of anatomical landmarks and grading articular cartilage in cadaveric knees. AB - PURPOSE: The purpose of the study was to define magnetic resonance imaging (MRI) findings before and after contrast medium opacification of the knee joint in cadaveric specimens to demonstrate anatomical landmarks of the trochlear surface in relation to the neighboring structures, and to evaluate different MRI sequences in the detection of cartilage defects of the trochlear and patellar surface of the knee. MATERIALS AND METHODS: The morphology and relationship of the proximal trochlear surface to the prefemoral fat of the distal femur were investigated by use of different MR sequences before and after intra-articular gadolinium administration into the knee joint in ten cadaveric knees. Anatomic sections were subsequently obtained. In addition, evaluation of the articular surface of the trochlea was performed by two independent observers. The cartilage surfaces were graded using a 2-point system, and results were compared with macroscopic findings. RESULTS: Of 40 cartilage surfaces evaluated, histopathologic findings showed 9 normal surfaces, 20 containing partial thickness defects, and 11 containing full-thickness defects. Compared with macroscopic data, sensitivity of MR sequences for the two reviewers was between 17 and 90%; specificity, 75 and 100%; positive predictive value, 75 and 100%; negative predictive value, 20 and 100%, depending on patellar or trochlea lesions. Interobserver variability for the presence of disease, which was measured using the kappa statistic, was dependent on the MR sequence used between 0.243 and 0.851. CONCLUSION: Magnetic resonance imaging sequences can be used to evaluate the cartilage of the trochlear surface with less accuracy when compared with the results of grading the articular cartilage of the patella. PMID- 18401582 TI - Concurrence of marginal zone B-cell lymphoma MALT-type and Langerhans cell histiocytosis in a thyroid gland with Hashimoto disease. PMID- 18401583 TI - Prolonged haematological toxicity from the hyper-CVAD regimen: manifestations, frequency, and natural history in a cohort of 125 consecutive patients. AB - The hyperfractionated cyclophosphamide, vincristine, doxorubicin and dexamethasone (hyper-CVAD) regimen has impressive efficacy in several haematological malignancies but is associated with considerable short-term haematological toxicity. Secondary myelodysplasia (MDS) or acute myeloid leukaemia (AML) also occurs. In this retrospective study, we also describe other prolonged haematological sequelae of this regimen. One hundred and twenty-five patients were treated with a median of six hyper-CVAD cycles and followed for a median of 28 months. Follow-up for cytopenias was censored at the next cytotoxic therapy. At 3 months post-therapy, 77 patients were evaluable. Cytopenias persisted in 59% of patients. Requirement for dose attenuation was the only factor significantly associated with persisting cytopenias (p<0.05). The median time to normalisation of counts for those with post-treatment cytopenias in the respective lineages was 9 months (range, 6-12) for anaemia, 6 months (range, 6 30) for neutropenia and 9 months (range, 6-30) for thrombocytopenia. MDS/AML was diagnosed in four patients at 4, 21, 24 and 37 months after therapy with a cumulative incidence rate of 4.43% at 4 years. These results indicate a considerable rate of prolonged haematological toxicity after hyper-CVAD and a modest rate of MDS at this limited follow-up. These findings likely reflect cumulative damage to haematopoietic stem cells. PMID- 18401584 TI - Absidia corymbifera necrotizing cellulitis in an immunocompromised patient while on voriconazole treatment. PMID- 18401585 TI - The face of high risk prostate cancer. AB - OBJECTIVES: To define high risk prostate cancer using prostate cancer specific mortality as the key outcome metric. METHODS: Data from two population based cohorts of men from Connecticut who were diagnosed with localized prostate cancer were analyzed to determine the natural history of prostate cancer and the impact of treatment on long term survival. RESULTS: Men with Gleason 7-10 prostate cancer and a potential survival of 10 years have a high risk of dying from their disease if they elect active surveillance. Surgery appears to offer an improved survival for these men when compared to radiation therapy or observation. Men diagnosed with Gleason 6 tumors in the contemporary era are more likely to harbor low risk prostate cancer when compared to historical series. CONCLUSIONS: Our studies confirm that high risk prostate cancer is best identified by Gleason score 7-10, but challenge the concept that men with high-grade disease are less likely to benefit from radical surgery. Men who have rising PSA values following treatment with either surgery or radiation have residual prostate cancer and are at very high risk of dying from prostate cancer within 10 years. PMID- 18401586 TI - Physical characteristics of digesta and their influence on flow and mixing in the mammalian intestine: a review. AB - The physical properties of digesta may influence mixing, efficiency of digestion, and absorption within the lumen of the intestine. We review how the physical properties of digesta change during transit through the various segments of the intestine, and how their influence on flow and mixing may be modulated by peristaltic activity. We examine how, in more fluid digesta, the solid and liquid phases interact to influence flow and mixing. Similarly, how in viscid digesta, shear strength, plasticity and elasticity of contained particulate material may influence the permeation of the fluid phase and secretions into and out of the digesta bolus. The manner in which the solid and liquid phases of digesta interact in a partly gaseous environment, such as the lower bowel, to influence bolus cohesion is also examined. Those mechanisms that promote the formation of a less viscous layer at the mucosal interface to promote plug flow are reviewed, and their effect on the efficiency of mixing and digestion discussed. It is recommended that in any future work investigating the character of mixing in the intestine, a wider range of appropriate digesta properties be measured and that, in investigations of intestinal movement, perfusates with similar characteristics to digesta be used. PMID- 18401587 TI - Congenital diaphragmatic hernia and retinoids: searching for an etiology. AB - Congenital diaphragmatic hernia (CDH) is a major life-threatening cause of respiratory failure in the newborn. Recent data reveal the role of a retinoid signaling pathway disruption in the pathogenesis of CDH. We describe the epidemiology and pathophysiology of human CDH, the metabolism of retinoids and the implications of retinoids in the development of the diaphragm and lung. Finally, we describe the existing evidence of a disruption of the retinoid signaling pathway in CDH. PMID- 18401588 TI - Baicalin protects human fibroblasts against ultraviolet B-induced cyclobutane pyrimidine dimers formation. AB - Exposure to ultraviolet B (UVB) irradiation is a major risk factor for the development of skin cancer. Therefore, it is important to identify agents that can offer protection against UVB-caused DNA damage. Photocarcinogenesis is caused largely by mutations at the sites of incorrectly repaired DNA photoproducts, of which the most common are the cyclobutane pyrimidine dimers (CPDs). In this study, a DNA damage model of UVB irradiation-induced fibroblasts was established. The immunocytochemical staining, immuno dot blotting and Western blotting were employed in the study. We demonstrated that pre-treatment of fibroblasts with Baicalin dose-dependently reduced the amount of UVB-generated CPDs. Compared with UVB irradiated cells, UVB-induced p53 accumulation was less pronounced in Baicalin-treated cells. Taken together, these results suggest that Baicalin prevent CPDs formation induced by UVB. Baicalin is therefore a promising protective substance against UVB radiation. PMID- 18401589 TI - Gas gangrene of the breast: management of a potential life-threatening infection. AB - INTRODUCTION: Gas gangrene of the breast is a rare infection and potentially mortal. CASE REPORT: We report a case of a fast extension of a painful right breast erythema whose starting point was a right parasternal cutaneous abscess. A diagnosis of gas gangrene of the right breast was made. A right mammectomy was carried out in Emergency and an antibiotherapy adapted to the germs was given. CONCLUSION: Mixed anaerobic and aerobic florae are often responsible for the infection. Its medico-surgical management is an emergency. PMID- 18401590 TI - Fetal therapy in twin reversed arterial perfusion sequence pregnancies with alcohol ablation or bipolar cord coagulation. AB - OBJECTIVE: We aimed to evaluate perinatal outcome of seven pregnancies with twin reserve arterial perfusion sequence. MATERIALS AND METHODS: The study group included seven cases of acardiac twins. Out of seven acardiac twins, two cases were in follow-up without any interventions. We performed four alcohol ablation and one bipolar coagulation. For alcohol ablation, a 20-gauge needle guided with color Doppler USG was directed to abdominal insertion site of the single umbilical artery of the acardiac twin, and 1.0-2.0 mL of absolute alcohol was injected. For bipolar coagulation of the umbilical cord, we used 3.5 mm laparoscopic trocar and 3.0 mm bipolar forceps. The procedures were performed under the guidance of transabdominal ultrasonography. RESULTS: Gestational age of the cases at diagnosis and at delivery was 15-32 and 17-38 weeks, respectively. Two cases without intervention were lost at 17 and 32 weeks. The mean time of procedure for bipolar coagulation and alcohol ablation were 30 and 10 min, respectively. One of the four cases of alcohol ablation group was aborted although alcohol ablation was successful. The other one case was aborted after alcohol ablation due to lost of fetal cardiac activity of the pump fetus. In two other cases, umbilical cord ablation with alcohol was successful, and they delivered live fetuses at 36 and 38 weeks. In one case we performed bipolar cord coagulation successfully, and the case delivered a live fetus at 39 weeks. The overall survival rate for intrauterine surgery was 60% (N=3/5). CONCLUSION: In twin reversed arterial perfusion sequence pregnancies with findings of poor prognosis, alcohol ablation or bipolar cord coagulation as fetal therapy under the guidance of ultrasonography can be done successfully, and should be offered as a choice to families upon discussion of intervention or follow-up with own complications. PMID- 18401591 TI - Surgical treatment and outcomes of temporal bone chondroblastoma. AB - Chondroblastoma is an uncommon primary benign bone tumor that usually arises in the epiphyses of the long bones. Temporal bone chondroblastoma is a rare primary bone tumor that affects the floor of the middle cranial fossa and temporomandibular joint (TMJ). The biological nature of temporal bone chondroblastoma is occasionally aggressive because of local invasion and is known to have a high recurrence after curettage. Therefore, complete resection is recommended. However, the literature provides little information regarding long term surgical outcomes and complications after surgical resection. The authors have retrospectively analyzed four cases of temporal bone chondroblastoma that had been completely excised by a single surgeon with an eventual long-term follow up. A single surgeon operated on four patients, two males and two females, with a mean age of 34 years, at the Department of Otorhinolaryngology, Severance Hospital. In all cases, the tumor involved the middle cranial fossa dura and the mandibular fossa with variable degree of infiltration. All patients have had no tumor recurrence to date (mean follow-up period of 5 years). Complete surgical resection of the temporal bone chondroblastoma is the gold standard for treatment. Precise preoperative image evaluation of tumor extension and proper management of the dura mater and temporomandibular joint (TMJ) are the major important features in complete surgical removal that minimize complications in temporal bone chondroblastoma treatment. PMID- 18401592 TI - Effects of endothelin-1 on calcium-independent contraction of bovine trabecular meshwork. AB - BACKGROUND: Endothelin-1 (ET-1) is known to induce contraction of trabecular meshwork (TM) and is probably involved in the pathogenesis of glaucoma. Calcium (Ca(2+))-independent contraction has been shown in TM, and its inhibition may represent an interesting way of influencing outflow facility, and thus intraocular pressure (IOP). This study investigates the role of ET-1 and its receptors ET-A and ET-B (ET-AR and ET-BR) in TM Ca(2+)-independent contractility. METHODS: Isometric tension measurements of bovine TM (BTM) strips were performed using a force-length transducer system. Intra- and extracellular Ca(2+) buffering was achieved by means of EGTA and BAPTA-AM. Under Ca(2+)-free conditions, ET-1 induced contractility of TM was assessed also in the presence of the specific inhibitors for ET-AR and ET-BR, BQ123 and BQ788 respectively. In order to clarify the intracellular mediators of Ca(2+)-independent contractility induced by ET-1, TM contraction was further measured in the presence of Y-27632, a selective inhibitor of Rho-associated kinases (ROCKs). The expression of ROCK1 and of its activating protein RhoA in BTM cells was investigated using western blot analysis. RESULTS: ET-1 induced a significant contraction of native BTM after intra- and extracellular Ca(2+)-depletion (45% +/- 8% of the maximally inducible contraction). Both endothelin receptor inhibitors BQ123 and BQ788 significantly reduced TM Ca(2+)-independent contraction in response to ET-1 (8.4 +/- 3.3% and 20.3 +/- 4.8% respectively). In the presence of the ROCK inhibitor Y-27632, ET-1 induced contraction of TM under Ca(2+)-free conditions was almost completely abolished (4.3% +/- 1.7%, p < 0.001). A clear signal for RhoA at 24 kDa and ROCK1 at 160 kDa could be detected in lysates of native tissue and cultured BTM cells with western blot. CONCLUSIONS: This study shows evidence that a significant portion of ET-1-induced contraction of TM is Ca(2+)-independent. In this contraction pathway, both ET-AR and ET-BR are involved with RhoA and its kinases as intracellular mediators. Ca(2+)-independent contraction of TM in response to ET-1 may represent a specific target to modulate IOP. PMID- 18401593 TI - A model of cardinality blindness in inferotemporal cortex. AB - A classifier is cardinality invariant if it can classify more than one token of a single type at a time. We present a convolutional neural network (CNN) model of inferotemporal cortex (IT) and show that it is cardinality invariant. While the CNN is designed with translation invariance in mind, cardinality invariance is an emergent property. We speculate that translation invariance may lead to cardinality invariance in general, and particularly in IT. Recent investigations have shown that cells in IT are indeed cardinality blind. We also explore the implications of a cardinality blind classifier for vision overall, concentrating on visual attention and search. PMID- 18401595 TI - Sex-steroidal regulation of aromatase mRNA expression in adult male rat brain: a quantitative non-radioactive in situ hybridization study. AB - Neuronal aromatase, the enzyme that catalyzes the conversion of androgens to estrogens, is involved in brain sexual differentiation, the regulation of reproductive behavior, and gonadotropin secretion. We have previously reported that aromatase P450 (AromP450) protein expression is enhanced by both androgens and estrogens in the principal nucleus of the bed nucleus of the stria terminalis (prBST) and posterodorsal part of the medial amygdaloid nucleus (pdMAm) of the adult rat but is not altered in the central amygdaloid nucleus (CeAm) even after sex-steroid withdrawal or supplementation. Here, we have evaluated, via in situ hybridization with digoxigenin-labeled cRNA probes, the sex-steroidal regulation of brain AromP450 mRNA in the prBST, pdMAm, and CeAm of orchidectomized and adrenalectomized adult male rats treated with sesame oil, testosterone (1 mg/rat/day), dihydrotestosterone (1 mg/rat/day), or 17beta-estradiol (2 microg/rat/day) for 6 days. AromP450-mRNA expression in the prBST and pdMAm was markedly reduced in orchidectomized/adrenalectomized rats treated with sesame oil but strongly enhanced by testosterone or dihydrotestosterone and significantly reinstated by 17beta-estradiol. These results are essentially consistent with those of AromP450 protein expression and thus indicate that enhanced AromP450 protein expression in the prBST and pdMAm reflects transcriptional upregulation and/or post-transcriptional stabilization of its mRNA by sex steroids. In the CeAm, despite moderate AromP450-protein expression, the mRNA has never been detected with or without sex-steroidal manipulations, indicating that the putative sex-steroid-insensitive AromP450 mRNA in the CeAm may be distinct from that in the prBST and pdMAm or, if it occurs at all, expressed at much lower levels. PMID- 18401594 TI - Genome-wide screen for asthma in Puerto Ricans: evidence for association with 5q23 region. AB - While the number of success stories for mapping genes associated with complex diseases using genome-wide association approaches is growing, there is still much work to be done in developing methods for such studies when the samples are collected from a population, which may not be homogeneous. Here we report the first genome-wide association study to identify genes associated with asthma in an admixed population. We genotyped 96 Puerto Rican moderate to severe asthma cases and 88 controls as well as 109 samples representing Puerto Rico's founding populations using the Affymetrix GeneChip Human Mapping 100K array sets. The data from samples representing Puerto Rico's founding populations was used to identify ancestry informative markers for admixture mapping analyses. In addition, a genome-wide association analysis using logistic regression was performed on the data. Although neither admixture mapping nor regression analysis gave any significant association with asthma after correction for multiple testing, an overlap analysis using the top scoring SNPs from different methods suggested chromosomal regions 5q23.3 and 13q13.3 as potential regions harboring genes for asthma in Puerto Ricans. The validation analysis of these two regions in 284 Puerto Rican asthma trios gave significant association for the 5q23.3 region. Our results provide strong evidence that the previously linked 5q23 region is associated with asthma in Puerto Ricans. The detection of causative variants in this region will require fine mapping and functional validation. PMID- 18401596 TI - Multipotent properties of myofibroblast cells derived from human placenta. AB - Human uterine fibroblasts (HuF) isolated from the maternal part (decidua parietalis) of a term placenta provide a useful model of in vitro cell differentiation into decidual cells (decidualization, a critical process for successful pregnancy). After isolation, the cells adhere to plastic and have either a small round or spindle-shaped morphology that later changes into a flattened pattern in culture. HuF robustly proliferate in culture until passage 20 and form colonies when plated at low densities. The cells express the mesenchymal cell markers fibronectin, integrin-beta1, ICAM-1 (CD54), and collagen I. Flow cytometry of HuF has detected the presence of CD34, a marker of the hematopoietic stem cell lineage, and an absence of CD10, CD11b/Mac, CD14, CD45, and HLA type II. Furthermore, they also express the pluripotency markers SSEA-1, SSEA-4, Oct-4, Stro-1, and TRA-1-81 as detected by confocal microscopy. Treatment for 14-21 days with differentiation-inducing media leads to the differentiation of HuF into osteoblasts, adipocytes, and chondrocytes. The presence of alpha smooth muscle actin, calponin, and myosin light-chain kinase in cultured HuF implies their similarity to myofibroblasts. Treatment of the HuF with dimethyl sufoxide causes reversion to the spindle-shaped morphology and a loss of myofibroblast characteristics, suggesting a switch into a less differentiated phenotype. The unique abilities of HuF to exhibit multipotency, even with myofibroblast characteristics, and their ready availability and low maintenance requirements make them an interesting cell model for further exploration as a possible tool for regenerative medicine. PMID- 18401597 TI - Resolving the three-dimensional myoarchitecture of bovine esophageal wall with diffusion spectrum imaging and tractography. AB - In order to determine the three-dimensional (3D) resolved muscular anatomy of the mammalian esophagus, we have examined its myoarchitecture with diffusion spectrum magnetic resonance imaging (DSI) and tractography. DSI measures diffusion displacement as a function of magnetic gradients of varied direction and intensity and displays the displacement profile as a 3D contour per voxel. In tractography, the orientation vectors of maximum diffusion/voxel are identified, and intervoxel associations are constructed by a streamline algorithm based on angular similarity in order to generate mesoscale myofiber tracts. We demonstrate that the proximal body of the esophagus consists of helically aligned crossing fiber populations that overlap between layers in the form of a "zipper" region along the length of the tissue. With increasingly distal position along the length of the tissue, helix angle and skeletal muscle prevalence are reduced such that fibers align themselves in the most distal location into distinct inner circular and outer longitudinal smooth muscle layers. We conclude that esophageal myoanatomy consists of crossing myofibers exhibiting a decreasing degree of helicity as a function of axial position and propose that this unique geometric construct provides a mechanism to resist distension and promote aboral flow. PMID- 18401599 TI - Subgroup analysis: at best suggestive (rather than conclusive). PMID- 18401600 TI - What is the value of magnetic resonance venography in children before renal transplantation? AB - Radiological evaluation before renal transplantation includes imaging of vascular anatomy, as thrombosis, narrowing and anomalies of the inferior vena cava (IVC) and/or iliac veins (IV) can influence the surgical technique. Most cases only require investigation with Doppler vessel ultrasonography (US), with magnetic resonance venography (MRV) reserved to clarify US findings and investigate high risk patients. The purpose of this study was to compare these modalities in evaluating IVC and IV and correlate imaging and operative findings of patients at RTx surgery. Twenty-nine children, 21 (72%) of whom had subsequent RTx surgery, were investigated over 5 years with correlation of US and MRV in 62% (18 of 29). Technically difficult US examinations needing MRV for clarification occurred in six (21%), and normal US with anatomical variations on MRV was seen in three (10%). The anatomical variations consisted of left-sided IVC, aberrant right common femoral vein and a left IV partly draining into the azygos and renal veins. US is an excellent screening tool for evaluating vascular anatomy patency in children. MRV infrequently contributes beneficial information, is difficult to justify as a screening tool, and due to the risks of gadolinium in uraemia, should only be used on an individual patient basis. PMID- 18401601 TI - [Minimally invasive injection treatment of radicular lumbal syndrome]. PMID- 18401603 TI - Knowledge and misconceptions of tuberculosis in the general population in Serbia. AB - This study was conducted in Serbia, including Vojvodina and central Serbia, but excluding Kosovo. We aimed to assess the level of knowledge and misconceptions about tuberculosis (TB) in the general population in Serbia. A total of 1,129 inhabitants aged at least 15 years old were interviewed. A combination of telephone and field surveys was used. More than 20% of respondents correctly understood the modes of transmission of TB. However, more than a quarter (28.5%) of respondents had some misconceptions about the possible modes of transmission. Analysis showed that the only significant predictors of a correct understanding of the modes of transmission were the education level and a close personal relationship with a patient with TB. The strongest predictors of misconceptions were older age and an urban type of settlement (odds ratio [OR] 1.5, confidence interval [CI] 1.3-1.6; OR 0.4, CI 0.3-0.6, respectively). A total of 33.9% of respondents could not identify any sign of TB and 86.6% of respondents knew that TB could be treated effectively, but only 20.1% knew that the treatment was free of charge. Our study showed that knowledge about TB is not satisfactory in the general population in Serbia. PMID- 18401602 TI - Postoperative pain following the use of two different intracanal medications. AB - This study evaluated the incidence of postoperative pain after intracanal dressings with either 0.12% chlorhexidine digluconate gel (CHX) or a calcium hydroxide/camphorated paramonochlorophenol/glycerin paste (CH/CPMC). Overall, 138 asymptomatic teeth had their canals instrumented under irrigation with 2.5% NaOCl and then dressed with either CH/CPMC or CHX. The incidence of different intensity levels of postoperative pain was registered for the period between appointments. Data revealed that 84% of the total number of cases treated with either medicament showed absence of any level of pain. No case medicated with CH/CPMC and four cases (5.8%) medicated with CHX were categorized as flare-ups. There were no statistically significant differences between all possible comparisons involving the two medicaments in treatment/retreatment cases and teeth with/without apical periodontitis lesions. The low incidence of postoperative pain after the use of both medications, coupled to their antimicrobial effectiveness, gives support to using one or the other in routine treatment/retreatment. PMID- 18401604 TI - Successful management of a ruptured hepatic artery pseudoaneurysm with a stent graft. AB - Hepatic artery pseudoaneurysm is a rare and potentially life-threatening vascular disorder. Careful consideration and early management of this lesion can be life saving. The authors report a case of a hepatic artery pseudoaneurysm which was successfully controlled by endovascular repair using a covered stent-graft. PMID- 18401605 TI - Pattern electroretinograms for the detection of neural loss in patients with permanent temporal visual field defect from chiasmal compression. AB - PURPOSE: To evaluate the ability of full-field and hemifield pattern electroretinogram (PERG) parameters to differentiate between healthy eyes and eyes with band atrophy (BA) of the optic nerve. METHODS: Twenty-six eyes from 26 consecutive patients with permanent temporal hemianopic visual field defects and BA of the optic nerve from previous chiasmal compression and 26 healthy subjects were studied prospectively. All patients were submitted to an ophthalmic examination including Humphrey 24-2 SITA Standard automated perimetry. Full-field and hemifield (nasal and temporal) stimulation transient pattern electroretinograms (PERG) were recorded using checkerboard screens. Amplitudes and peak times for the P50 and N95 as well as the overall P50+N95 amplitude were measured. The intraocular N95:P50 amplitude ratio was calculated. Comparisons were made using Student's t-test. Receiver operating characteristic (ROC) curves were used to describe the ability of PERG parameters to discriminate the groups. RESULTS: Full-field P50, N95, and P50+N95 amplitude values were significantly smaller in eyes with BA than in control eyes (P < 0.001). Nasal and temporal hemifield PERG studies revealed significant differences in N95 and P50+N95 amplitudes measurements. No significant difference was observed regarding peak times or N95:P50 amplitude ratios. Nasal and temporal hemifield PERG values did not differ significantly in eyes with BA or in controls. Using the 10th percentile of normals as the lower limit of normal, 16 of 26 eyes were considered abnormal according to the best discriminating parameters. CONCLUSIONS: Transient PERG amplitude measurements were efficient at differentiating eyes with BA and permanent visual field defects from normal controls. Hemifield stimulation PERG parameters were unable to detect asymmetric hemifield neural loss, but further studies are required to clarify this issue. PMID- 18401609 TI - Cardiac MR imaging: from physics to protocols. PMID- 18401610 TI - Pediatric cardiac MRI: anatomy and function. PMID- 18401611 TI - Cardiac CTA: congenital heart disease. PMID- 18401612 TI - New perspectives in imaging interstitial lung disease in children. PMID- 18401613 TI - Advancing CT and MR imaging of the lungs and airways in children: imaging into practice. PMID- 18401614 TI - Sedating children for radiological procedures: an intensivist's perspective. PMID- 18401615 TI - The pediatric sedation service: who is appropriate to sedate, which medications should I use, who should prescribe the drugs, how do I bill? PMID- 18401616 TI - Avoiding sedation in pediatric radiology. PMID- 18401617 TI - Pediatric DXA: technique, interpretation and clinical applications. PMID- 18401618 TI - MR arthrography made simple: indications and techniques. PMID- 18401619 TI - Musculoskeletal imaging at 3 T: imaging and optimization. PMID- 18401620 TI - Musculoskeletal imaging: evaluation of congenital anomalies. PMID- 18401621 TI - Major pitfalls in musculoskeletal imaging-MRI. PMID- 18401622 TI - Cartilage imaging. PMID- 18401623 TI - Pediatric abdominal CT angiography. AB - Advancing multidetector technology offers opportunities for improved vascular assessment in children. Much of what is available deals with thoracic and central nervous system applications, with very little written about abdominal applications. That said, many of the technical aspects are similar to computed tomography (CT) angiography in these regions and are worthy of reviewing, in addition to those unique considerations for abdominal CT angiography (CTA) in children. Familiarity with appropriate abdominal CTA for pediatric multidetector array CT will provide the same opportunities as CTA in other regions. PMID- 18401624 TI - Pancreatic and biliary anomalies: imaging in 2008. PMID- 18401625 TI - Imaging of the bowel in children: new imaging techniques. PMID- 18401626 TI - MR urography: anatomy and physiology. PMID- 18401627 TI - Contrast media toxicity in children. PMID- 18401631 TI - An unusual cause of persistent headache: chloroma (2008: 2b). AB - A chloroma (granulocytic sarcoma) is a rare tumour that is usually associated with leukaemia. The osseous skeleton is most commonly involved, and it confers a poorer prognosis for the underlying leukaemic disease. We present a case of a chloroma of the nasal sinuses that was the primary presentation of the patient's underlying leukaemia. PMID- 18401633 TI - An extensive study of mutation and selection on the wobble nucleotide in tRNA anticodons in fungal mitochondrial genomes. AB - Two alternative hypotheses aim to predict the wobble nucleotide of tRNA anticodons in mitochondrion. The codon-anticodon adaptation hypothesis predicts that the wobble nucleotide of tRNA anticodon should evolve toward maximizing the Watson-Crick base pairing with the most frequently used codon within each synonymous codon family. In contrast, the wobble versatility hypothesis argues that the nucleotide at the wobble site should be occupied by a nucleotide most versatile in wobble pairing, i.e., the wobble site of the tRNA anticodon should be G for NNY codon families and U for NNR and NNN codon families (where Y stands for C or U, R for A or G, and N for any nucleotide). We examined codon usage and anticodon wobble sites in 36 fungal genomes to evaluate these two alternative hypotheses and identify exceptional cases that deserve new explanations. While the wobble versatility hypothesis is generally supported, there are interesting exceptions involving tRNA(Arg) translating the CGN codon family, tRNA(Trp) translating the UGR codon family, and tRNA(Met) translating the AUR codon family. Our results suggest that the potential to suppress stop codons, the historical inertia, and the conflict between translation initiation and elongation can all contribute to determining the wobble nucleotide of tRNA anticodons. PMID- 18401634 TI - Effectiveness of angiotensin-converting enzyme inhibitors in pediatric patients with mid to severe aortic valve regurgitation. AB - The long-term benefit of angiotensin-converting enzyme inhibitors in pediatric patients with aortic valve regurgitation is under consideration. Eighteen patients with mid to severe aortic valve regurgitation were retrospectively evaluated. Echocardiographic parameters (left ventricular end-diastolic diameter, shortening fraction, left ventricular posterior wall thickness, and grade of aortic valve regurgitation) were analyzed before and during therapy with angiotensin-converting enzyme inhibitors. Data are given as standard deviation scores (Z-scores) derived from body surface-adjusted normal values. Median (interquartile range) age at start of therapy was 8.4 (5.4 to 10.0) years, and total follow-up 2.3 (0.9 to 5.4) years. Left ventricular end-diastolic diameter increased from 3.6 (2.3 to 4.5) to 3.7 (2.4 to 4.8), and left ventricular posterior wall diameter decreased from 1.9 (1.1 to 3.0) to 1.1 (0.5 to 2.3). Grade of aortic valve regurgitation increased from 3.5 (2.3 to 4.0) to 4.0 (2.0 to 4.0), and shortening fraction decreased from 39% (34% to 43%) to 37% (34% to 42%). No significant effect of angiotensin-converting enzyme inhibitors on left ventricular dimensions or function was found in our population of patients with mid to severe aortic valve regurgitation. Angiotensin-converting enzyme inhibitors may not alter left ventricular overload in pediatric patients with aortic valve regurgitation. PMID- 18401635 TI - A single center's experience with the Ross procedure in pediatrics. AB - The use of a pulmonary autograft for aortic valve replacement (AVR) has become more prevalent than other forms of AVR in the pediatric population. We reviewed the data on pediatric patients who underwent the Ross procedure at our institution from 1993 to 2005. Sixty patients <18 years old who underwent a Ross procedure had available clinical and echocardiographic data collected and statistical analysis performed. Mortality rate was 3.3%, while overall survival and freedom from reoperation of either the homograft or the autograft were 96.7% and 66.2%, respectively, at 10 years. Freedom from reoperation of the left ventricular outflow tract was 60.5% at 10 years. Echocardiographic data showed aortic regurgitation to be mild or less in 76% of patients by last follow-up, while dilation of the sinuses of Valsalva had occurred in 52%. Compared to other AVR options, the Ross procedure in eligible pediatric patients demonstrates good intermediate survival rates and continued growth potential, yet a time-dependent need for reoperation. PMID- 18401636 TI - Indomethacin therapy for patent ductus arteriosus in premature infants. PMID- 18401637 TI - Assessing and prioritizing ecological communities for monitoring in a regional habitat conservation plan. AB - In nature reserves and habitat conservation areas, monitoring is required to determine if reserves are meeting their goals for preserving species, ecological communities, and ecosystems. Increasingly, reserves are established to protect multiple species and communities, each with their own conservation goals and objectives. As resources are always inadequate to monitor all components, criteria must be applied to prioritize both species and communities for monitoring and management. While methods for prioritizing species based on endangerment or risk have been established, approaches to prioritizing ecological communities for monitoring are not well developed, despite a long-standing emphasis on communities as target elements in reserve design. We established guidelines based on four criteria derived from basic principles of conservation and landscape ecology--extent, representativeness, fragmentation, and endangerment--to prioritize communities in the San Diego Multiple Species Conservation Plan (MSCP). The MSCP was one of the first multiple-species habitat conservation areas established in California, USA, and it has a complex spatial configuration because of the patterns of surrounding land use, which are largely urbanized. In this case study, high priority communities for monitoring include coastal sage scrub (high endangerment, underrepresented within the reserve relative to the region, and moderately fragmented), freshwater wetlands, and coastal habitats (both have high fragmentation, moderate endangerment and representativeness, and low areal extent). This framework may be useful to other conservation planners and land managers for prioritizing the most significant and at-risk communities for monitoring. PMID- 18401638 TI - A misleading meta-analysis of seprafilm. PMID- 18401639 TI - Parathyroid cyst: often mistaken for a thyroid cyst. PMID- 18401640 TI - What is a surgical complication? PMID- 18401641 TI - Can surgeons assess CT suitability for endovascular repair (EVAR) in ruptured abdominal aortic aneurysm? Implications for a ruptured EVAR trial. AB - The purpose of this study was to determine whether surgeons without formal radiological training are able to assess suitability of patients with ruptured abdominal aortic aneurysms (AAA) for EVAR. The CT scans of 20 patients with AAA were reviewed under timed conditions by six vascular surgeons. Twenty minutes was allocated per scan. They were asked to determine if each aneurysm would be treatable by EVAR in the emergency setting and, if so, to measure for device selection. The results were then compared with those of a vascular radiologist. Six surgeons agreed on the suitability of endovascular repair in 45% of cases (95% CI, 23.1-68.5%; 9/20 scans; kappa = 0.41 [p = 0.01]) and concurred with the radiologist in eight of these. Individually, agreement ranged from 13 to 16 of the 20 scans, 65-80% between surgeons. The kappa value for agreement between all the surgeons and the radiologist was 0.47 (p = 0.01, moderate agreement). For the individual surgeons, this ranged from 0.3 to 0.6 (p = 0.01). In conclusion, while overall agreement was moderate between the surgeons and the radiologist, it is clear that if surgeons are to assess patients for ruptured EVAR in the future, focused training of surgical trainees is required. PMID- 18401642 TI - Circulating endothelial cells and endothelial progenitor cells in obstructive sleep apnea. AB - Increased circulating endothelial cells (CECs) have been observed in patients with vascular injury associated with acute myocardial infarction, pulmonary hypertension, and congestive heart failure. Decreased circulating endothelial progenitor cells (EPCs) have been observed in patients with risk factors for cardiovascular disease. Obstructive sleep apnea (OSA) is associated with increased risk of cardiovascular disease and endothelial dysfunction. Subjects were recruited from patients referred for overnight polysomnograms; 17 subjects had OSA and 10 control subjects did not have OSA. All subjects lacked vascular disease and risk factors for vascular disease. Peripheral blood was obtained from fasting subjects in the morning, following sleep studies. CECs and EPCs were quantified using magnetic bead separation with UV epifluorescence microscopy and flow cytometry immunophenotyping, respectively. Cell counts and demographic variables were compared using unpaired t tests. Regression analysis was performed comparing cell counts with the apnea-hypopnea index (AHI) and nadir SaO(2). Subjects with OSA and controls did not differ significantly in terms of age and body mass index. Subjects with OSA had higher AHI, lower nadir SaO(2), and greater sleepiness (Epworth Sleepiness Scale scores). There were no significant differences in CEC (7.0+/-1.5 vs. 4.9+/-0.9, p>0.05) or EPC (1077+/-318 vs. 853+/ 176, p>0.05) between controls and OSA cases, respectively. In this small study, we found no differences in CECs or circulating EPCs between patients with OSA and controls. OSA may not be associated with these markers of vascular endothelial cell injury in patients with no concomitant vascular disease. PMID- 18401643 TI - Influence of learning curve on short-term results after laparoscopic resection for rectal cancer. AB - BACKGROUND: Technical difficulties have been encountered in laparoscopic surgery for the treatment of rectal cancer. There are fewer studies about the learning curve for laparoscopic rectal resection. METHODS: Between June 1995 and August 2007, 200 patients who were scheduled to undergo laparoscopic rectal resection for rectal cancer were enrolled in the study. Each surgeon's operative experience was divided into three groups: 1-20 cases, 21-40 cases, and 41 or more cases. Furthermore, patients were divided chronologically into four groups of 50 patients each. This report describes the association between the learning curves (surgeon's experience and team's experience) and short-term outcomes such as operating time, complication rate, and hospital stay in the case of laparoscopic resection for rectal cancer. We also analyzed how the learning curve influences several postoperative outcomes compared with other clinical factors. RESULTS: The team's experience was not associated with short-term results except for surgical site infection (SSI). On the other hand, surgeon's experience was associated with mean operating time and SSI rate. The endpoints of the learning curve for reducing mean operating time and SSI rate were defined as 40 and 20 cases of laparoscopic rectal resection. In contrast, anastomotic leakage was not associated with surgeon's experience and showed the greatest correlation with total mesorectal excision (TME). CONCLUSION: Surgeon's learning improved operating time and SSI. On the other hand, low level of anastomosis accompanied with TME was strongly related with leakage, and the association between leakage and surgeon's learning was not clearly demonstrated. PMID- 18401644 TI - The role of video-assisted laparoscopy in management of patients with small bowel injuries in abdominal trauma. AB - BACKGROUND: Patients with small bowel injuries (SBI) in abdominal trauma have no clear clinical or radiological signs on initial examination. This leads to delay in appropriate surgical interventions with consequent high morbidity and mortality. In this paper we demonstrate the role of video-assisted laparoscopy (VAL) in management of such patients. METHODS AND MATERIALS: 819 patients with SBI were evaluated retrospectively between 1994 and 2003. The mechanism of the trauma was blunt in 146 (17.8%) patients and stab wounds in 640; routine investigations and VAL were used for triage of the subjects. All the victims underwent laparotomy or video-assisted laparoscopy. Demographic information, incidence of organs injuries, types of surgery, complications, associated injuries, and hospital mortality were analyzed. RESULTS: Diagnostic video laparoscopy was sufficient for 518 (63.2%) patients. Small bowel repair was performed in 332 (40.6%) patients, clipping of mesenteric blood vessels in 27 (3.4%) patients, and coagulation and ligation of omental blood vessels in 48 (5.9%) patients. Ninety-seven (11.8%) patients with small bowel injury were associated with postoperative complications. There were 19 (2.3%) deaths associated with hemoperitoneum, severe head injury, and multiple ribs fracture. Generalized peritonitis was revealed in one case and focal abscesses between intestinal loops were identified in another one patient. CONCLUSION: The DVAL findings of visceral injuries give optimal approach for management of small bowel injuries in abdominal trauma. PMID- 18401645 TI - The risk of communicating TEP-related infertility risk is an opportunity and not a "Cinderella concern" any more. PMID- 18401646 TI - The fate of patients who undergo "preoperative" ERCP to clear known or suspected bile duct stones. AB - BACKGROUND: There is debate as to whether recurrent biliary complications are more common in patients who do not have elective cholecystectomy after endoscopic retrograde cholangiopancreatography (ERCP) management of common bile duct (CBD) stones. The aim of this study was to determine the fate of patients with intact gallbladders who have had CBD stones removed at ERCP, and to assess their risk of recurrent biliary symptoms. METHODS: We retrospectively identified all patients in our large tertiary center population with intact gallbladders who had an ERCP for CBD stones from December 1999 to March 2002. We determined which patients had subsequent elective cholecystectomy, and the outcomes of patients who did not have elective surgery. RESULTS: 309 patients had CBD stones at ERCP during the study period, of which 139 had intact gallbladders at the time of ERCP. Of these 139 patients 59 had subsequent elective cholecystectomy, 11 by open operation and 48 laparoscopically. Of these 139 patients, 27 had cholecystectomy planned; 47 patients were managed with a wait-and-see strategy, 30 of whom were poor surgical candidates. Of these 47 patients in whom a wait-and-see policy was adopted, 9 (19%) developed complications including recurrent pain and/or abnormal liver function tests (LFTs), recurrent biliary colic, and pancreatitis. Eight of these nine patients were from the poor surgical candidate group. Sphincterotomy had been performed at initial ERCP in all patients. CONCLUSIONS: Over half of our population of 139 patients with CBD stones at ERCP and intact gallbladders had actual or planned elective cholecystectomy. For those patients in whom a decision to wait-and-see was made, almost 20% developed complications. Elective cholecystectomy after a finding of choledocholithiasis is supported by many and is a common strategy in our experience. Recurrent biliary complications are relatively common in those who do not undergo elective cholecystectomy, especially those patients who represent a high operative risk. PMID- 18401647 TI - Laparoscopic total colectomy for colonic inertia: surgical and functional results. AB - BACKGROUND: Total colectomy is the surgery of choice for colonic inertia (CI) when medical treatment has failed. Laparoscopic total colectomy has demonstrated to be a feasible technique. OBJECTIVE: Present our experience in patients with CI who underwent laparoscopic total colectomy (TC) + ileorectoanastomosis (IRA) and evaluate the functional results and medium-term follow-up after this surgery. MATERIAL AND METHODS: All patients with CI were included in a prospective laparoscopic surgical protocol, from 2002 to 2007. These patients had a complete work-up for chronic constipation (clinical records, barium enema, colonic transit time, defecography, anorectal manometry, small bowel follow through). All patients were evaluated with Wexner's score for constipation pre- and postoperatively, asked if they would recommend surgery to other patients, and if they were satisfied with the procedure (on a scale from 1 to 10). Statistical analysis was carried out using Student's T-test. RESULTS: In this period 20 patients were operated with diagnosis of CI. All patients were females with an average age of 41.5 years (range 18-52 years). Mean operative time was 248 min (range 170-360 min). One (5%) patient was converted to open surgery. The medium time to flatus per anum and feeding was 2 (range 1-6) and 3 (range 2-6) days, respectively. The medium postoperative stay was 7 days. Seven patients (35%) presented surgical postoperative complications (three postoperative ileus, one portal thrombosis, one rectal hemorrhage, one anastomotic leakage, and one hemoperitoneum). There was no postoperative mortality. The average follow-up was 25 months (range 1-60 months). Preoperative Wexner's constipation score was 22.3 (range 19-29 months) pre surgery and at the end of follow-up was 1.8 (range 0-6) (p < 0.01). The medium level of satisfaction was 8 (range 2-10) and only one patient would not recommend surgery to other patients. CONCLUSION: The laparoscopic access is a safe technique with satisfactory functional results after medium-term follow-up. PMID- 18401648 TI - Validated robotic laparoscopic surgical training in a virtual-reality environment. AB - BACKGROUND: A robotic virtual-reality (VR) simulator has been developed to improve robot-assisted training for laparoscopic surgery and to enhance surgical performance in laparoscopic skills. The simulated VR training environment provides an effective approach to evaluate and improve surgical performance. This study presents our findings of the VR training environment for robotic laparoscopy. METHODS: Eight volunteers performed two inanimate tasks in both the VR and the actual training environment. The tasks were bimanual carrying (BC) and needle passing (NP). For the BC task, the volunteers simultaneously transferred two plastic pieces in opposite directions five times consecutively. The same volunteers passed a surgical needle through six pairs of holes in the NP task. Both tasks require significant bimanual coordination that mimics actual laparoscopic skills. Data analysis included time to task completion, speed and distance traveled of the instrument tip, as well as range of motion of the subject's wrist and elbow of the right arm. Electromyography of the right wrist flexor and extensor were also analyzed. Paired t-tests and Pearson's r were used to explore the differences and correlations between the two environments. RESULTS: There were no significant differences between the actual and the simulated VR environment with respect to the BC task, while there were significant differences in almost all dependent parameters for the NP task. Moderate to high correlations for most dependent parameters were revealed for both tasks. CONCLUSIONS: Our data shows that the VR environment adequately simulated the BC task. The significant differences found for the NP task may be attributed to an oversimplification in the VR environment. However, they do point to the need for improvements in the complexity of our VR simulation. Further research work is needed to develop effective and reliable VR environments for robotic laparoscopic training. PMID- 18401649 TI - Liver resection with intraoperative and laparoscopic ultrasound: report of 32 cases : Ultrasonic shears device for liver parenchymal transection. AB - BACKGROUND: Careful staging of hepatic tumors is mandatory for appropriate selection of patients for liver resection. Number and relationships of liver nodules are issues of utmost importance when evaluating resectability. Sensitivity of preoperative imaging for secondary lesions has been reported between 60-75% with spiral contrast-enhanced computed cosmography (CT), 80-85% with magnetic resonance imaging (MRI), and 90-96% with intraoperative ultrasound (IOUS). Also for primary lesions IOUS has been reported to allow detection of liver nodules in 17% and modify surgical strategy in 10%. The aim of this study was to point out the usefulness of open (IOUS) and laparoscopic (LIOUS) ultrasound in patients undergoing hepatic surgery for liver tumors. METHODS: In the years 2004-2006, 50 patients, mean age 66 years (range 44-76 years) were evaluated for resective surgery at the General Surgery Department of Monaldi Hospital, Naples, Italy. All of them were studied with biphasic CT and transabdominal ultrasound. Eighteen (36%) were judged unresectable. The others were scheduled for laparoscopy and LIOUS, by means of an ALOKA SSD-5500 (Aloka Co. Ltd. Tokyo, Japan), equipped with linear flexible tip laparoscopic probe. RESULTS: Six patients (18.7%) were excluded because of pathology diffusion; 26 (81.3%) were resected, using ultrasonic shears (Harmonic ACE, Ethicon Endo Surgery, Cincinnati Oh., USA) for parenchymal transection, 3 (11.5%) laparoscopically and 23 (88.5%) after laparotomy. IOUS was repeated in the latter group. LIOUS spared useless laparotomies in six patients (18.7%) and, coupled with IOUS, found undetected nodes in five patients (19.2%), changing surgical strategy in three patients (11.5%). CONCLUSION: In our experience LIUOS and IOUS proved to be of utmost importance both in the selection of patients for resective surgery and in planning surgical approach. Ultrasonic shears device and systematic pedicle clamping sped up resection time and reduced intraoperative bleeding. PMID- 18401650 TI - Assessing patient-reported outcomes of cholecystectomy in short-stay surgery. AB - BACKGROUND: The project aimed at testing the feasibility of a quality improvement system based on patient-reported outcomes in short-stay surgery for cholecystectomy. METHODS: In two centers for short-stay surgery, patients referred for laparoscopic cholecystectomy were surveyed between Aug 1999 and Jan 2002. Patients reported on health-related quality of life (SF-36), symptoms (gallstone symptom checklist, GSCL) and other indicators preoperatively (T0), 14 days (T1) and 6 months postoperatively (T2). The SF-36 subscales physical functioning, bodily pain, and role physical as well as the GSCL and treatment satisfaction at T2 were considered as main outcomes. The main outcomes were analyzed by generalized linear models with regard to predictors. RESULTS: At T0 a total of 205 patients were included. The response rate at T2 was 63.4% (74.6% females, 53.6 years of age). The GSCL score at T0 (29.4% preoperatively) fell slightly to 27.9% at T1 and decreased to 14.8% at T2 (T0-T2: p < 0.001). The SF 36 subscales showed a different course over time (decrease of health-related quality of life at T1 and large increase at T2). The level of satisfaction with the outcome of cholecystectomy was 82.3%. The patient-reported outcomes were mainly influenced by the preoperative level, age and self-reported postoperative complaints. CONCLUSION: The low response rate was mainly due to nondelivery of questionnaires at T1 during the regular postoperative visit by the operating physician. Though nonresponse occurs under conditions of routine care, meaningful information was gained, which should be used for quality improvement activities. Because the preoperative level is a major determinant of the postoperative health outcomes, the prospective pre-post measurement should be preferred if institutional comparisons are intended. PMID- 18401651 TI - Endoscopic characteristics of gastric adenomas suggesting carcinomatous transformation. AB - BACKGROUND: Currently, endoscopic resections, including endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD), are widely performed for the management of gastric neoplasia. This study aimed to evaluate the potential predictive factors for carcinomas on the basis of endoscopic features. METHODS: This study investigated 114 samples from 114 patients. Gastric adenoma was diagnosed initially for all the patients. The endoscopic findings were reviewed for location, size, gross appearance, surface nodularity, ulceration, surface color, and number of biopsy samples. These variables were analyzed and compared between an adenoma group (51 cases) and a carcinoma group (63 cases) on the basis of postresection diagnosis. RESULTS: The mean age of the patients was 62 years (range, 43-82 years), and 83 of the patients were men. The diameter of the lesions was 14.6 +/- 8.2 mm in the adenoma group and 15.4 +/- 7.4 mm in the carcinoma group. Depressed type, combined high-grade dysplasia, red discoloration, and mucosal ulceration were significant variables associated with carcinomas. In the multivariate analysis, combined high-grade dysplasia was a significant independent predictor of carcinomas. CONCLUSIONS: The results suggest that patients with high-grade dysplasia on forceps biopsies should be considered candidates for endoscopic resection. Characteristics of gastric adenomas such as a depressed type, red color, and ulceration that may have foci of carcinomas in other parts of the adenomas also should be considered for endoscopic resection. PMID- 18401652 TI - What a signature adds to the consent process. AB - BACKGROUND: "Consent is a process by which a patient is informed and becomes a participant in decisions regarding their medical management." It is argued, however, that providing a signature to a form adds little to the quality of this process. METHODS: Views regarding the consent ritual of nonselected patients undergoing endoscopy (cystoscopy or sigmoidoscopy) were prospectively studied together with those of the attending staff. Patient volunteers were randomly assigned to one of two groups and given verbal explanation before the procedure, either alone (group A) or with a request to sign a form in addition (group B). A standardized questionnaire regarding preferences then was applied. RESULTS: A total of 37 patients (22 men) were studied along with seven staff members. Most surveyed felt that signing a consent form helped to empower the patient (group A, 84%; group B, 83%; staff, 100%). Although the patients mainly believed that it functioned primarily to protect the hospital and doctor (group A, 89%; group B, 67%), only one patient (3% of total) felt that such a formality undermined the patient-doctor relationship. Most staff members favored signing a form (86%). The majority of patients either favored it (group A, 47%; group B, 78%) or expressed no strong preference (group A, 32%; group B, 11%). Interestingly, more women than men preferred signing (73 vs. 55%; p = 0.25), perhaps because more women believed that it functioned to preserve autonomy (93 vs. 77% of men). Age was no particular determinant of perspective. CONCLUSION: Although it may be viewed as primarily serving to protect the doctor and hospital, the formal process of signing written consent forms appeals to patients and staff. PMID- 18401654 TI - Laparoscopic gastrectomy: advances enable wide clinical application. PMID- 18401655 TI - The role of three-dimensional endoluminal ultrasound imaging in the evaluation of anorectal diseases: a review. AB - BACKGROUND: The authors conducted a review of the available English literature to evaluate the advantages of three-dimensional ultrasound for assessing anorectal pathology, to provide a state-of-the-art approach, and to compare this technique with conventional endoluminal ultrasound and other imaging methods. METHODS: All studies describing results obtained with three-dimensional ultrasound in the evaluation of anorectal pathologies, both alone and compared with other techniques, were selected. RESULTS: Since 1996, 32 articles have been published. In tumor staging, specific data derived by three-dimensional reconstruction for the assessment of T invasion and nodal involvement were more accurate than endoluminal ultrasound and computed tomography. For evaluating perianal sepsis, the detection of secondary fistula tracts and fluid collections and the location of internal openings were superior to endoanal magnetic resonance imaging. In the evaluation of anal incontinence, the results for sphincter defects were similar to those obtained with endoanal magnetic resonance imaging. CONCLUSION: Analysis of the literature confirmed that three-dimensional ultrasound is a valuable technique for assessing anorectal disorders, facilitating the interpretation of the images obtained, and providing additional data that in many cases have changed the operative approach. PMID- 18401653 TI - Laparoscopic entry techniques: clinical guideline, national survey, and medicolegal ramifications. AB - BACKGROUND: This study aimed to establish criteria for safe laparoscopic entry through a systematic literature search and evidence-based medicine appraisal, to determine surgeon preferences for laparoscopic entry in the United Kingdom, and to appraise the medicolegal ramifications of complications arising from laparoscopic entry. METHODS: A systematic literature search of MEDLINE and EMBASE (1996-2007) was performed as well as a national surgeon survey by questionnaire (May-December 2006). RESULTS: Laparoscopic entry criteria involving 10 steps were established based on the systematic literature search and evidence-based critical appraisal. The national survey had 226 respondents, with the majority aware of the Middlesbrough consensus or Royal College of Obstetricians and Gynaecologists [RCOG]-sourced guidance. There was considerable variation in preferred laparoscopic entry techniques. Currently, there is clear judicial guidance on the medicolegal stance toward laparoscopic entry-related complications. CONCLUSIONS: Despite widespread awareness of laparoscopic entry guidelines, there remains considerable variation in the techniques adopted in clinical practice. Unless practice concurs with recommended guidance, women undergoing laparoscopy will be exposed to increased unnecessary operative risk. Laparoscopic entry-related injury in an uncomplicated woman is considered negligent practice according to UK legal case law. PMID- 18401656 TI - Challenges of rural surgery. PMID- 18401657 TI - Advanced endoscopic technologies. AB - The use of endoscopy for diagnosing and treating ailments of the alimentary tract has evolved steadily over the past few decades, with tremendous growth and innovation in the past few years. Initially, endoscopy relied on rigid telescopes, direct visualization, and dangerously exothermic sources of illumination. The introduction of fiber optics, charge-coupled-device cameras, and increasingly efficient light sources has enabled researchers to investigate areas of the human gastrointestinal tract through flexible endoscopy not previously thought to be reachable without formal surgical exploration. The more recent advances in scope platforms, devices, and techniques have allowed researchers to push the envelope of endoscopic diagnostics and therapeutics to greater heights. Specific new platforms include ColonoSight and mother-daughter endoscopes such as the ShapeLock TransPort and the SpyGlass direct visualization system. Specific devices include the EndoCinch suturing system, the full thickness Plicator procedure, Esophyx, the Stretta system, and the HALO(360) system. Specific new techniques include small-caliber endoscopy, endoscopic mucosal and submucosal resection, and natural orifice translumenal endoscopic surgery (NOTES). This article describes the most relevant recent advances in endoscopic innovation with regard to platform design, devices, and techniques anticipated to serve as the foundation for further research and design for developing generations of endoscopic technologies to come. PMID- 18401658 TI - Reliable gastric closure after natural orifice translumenal endoscopic surgery (NOTES) using a novel automated flexible stapling device. AB - BACKGROUND: Reliable closure of the translumenal incision is one of the main challenges facing natural orifice translumenal endoscopic surgery (NOTES). This study aimed to evaluate the use of an automated flexible stapling device (SurgASSIST) for closure of the gastrotomy incision in a porcine model. METHODS: A double-channel gastroscope was advanced into the stomach. A gastric wall incision was made, and the endoscope was advanced into the peritoneal cavity. After peritoneoscopy, the endoscope was withdrawn into the stomach. The SurgASSIST stapler was advanced orally into the stomach. The gastrotomy edges were positioned between the opened stapler arms using two endoscopic grasping forceps. Stapler loads with and without a cutting blade were used for gastric closure. After firing of the stapler to close the gastric wall incision, x-ray with contrast was performed to assess for gastric leakage. At the end of the procedure, the animals were killed for a study of closure adequacy. RESULTS: Four acute animal experiments were performed. The delivery and positioning of the stapler were achieved, with technical difficulties mostly due to a short working length (60 cm) of the device. Firing of the staple delivered four rows of staples. Postmortem examination of pig 1 (when a cutting blade was used) demonstrated full-thickness closure of the gastric wall incision, but the cutting blade caused a transmural hole right at the end of the staple line. For this reason, we stopped using stapler loads with a cutting blade. In the three remaining animals (pigs 2-4), we were able to achieve a full-thickness closure of the gastric wall incision without any complications. CONCLUSIONS: The flexible stapling device may provide a simple and reliable technique for lumenal closure after NOTES procedures. Further survival studies are currently under way to evaluate the long-term efficacy of gastric closure with the stapler after intraperitoneal interventions. PMID- 18401659 TI - A modified method of laparoscopic side-to-side esophagojejunal anastomosis: report of 14 cases. AB - BACKGROUND: Alimentary tract reconstruction after laparoscopic total gastrectomy is a technical challenge. Although feasible, reconstruction through a small incision has several drawbacks. The authors therefore report a modified method of laparoscopic side-to-side esophagojejunal anastomosis developed at their hospital. METHODS: The side to side esophagojejunal anastomosis was completed with a endo-GIA firing, followed by transection of the jejunum and esophagus with another firing of endo-GIA. RESULTS: This modified procedure was performed successfully for 14 patients with gastric cancer. The mean operation time for this procedure was 42.5 +/- 10.2 min. No postoperative death, fistula, or hemorrhage occurred. All the patients were followed up for a mean period of 14.5 months with no cancer recurrence at the anastomosis or anastomotic stricture. All the patients had a barium swallow test 6-2 months after the operation. The mean maximum diameter of the anastomosis was 3.8 cm (range, 3.0-4.2 cm). Four patients experienced temporary symptoms of dumping syndrome or dysphagia, which disappeared 6 months postoperatively. CONCLUSION: The authors consider this modified laparoscopic side-to-side esophagojejunal anastomosis to be safe, less challenging, and more economical, providing an alternative for alimentary tract reconstruction after laparoscopic total gastrectomy. PMID- 18401660 TI - Development of tolerance to the dietary plant secondary metabolite 1,8-cineole by the brushtail possum (Trichosurus vulpecula). AB - The common brushtail possum (Trichosurus vulpecula) is a generalist herbivore whose diet includes Eucalyptus leaves that are well defended by plant secondary metabolites (PSM) such as the terpene 1,8-cineole (cineole). We accustomed possums to a terpene-free diet, then challenged them with the addition of 2% cineole to the diet. Initially, there was a 50% reduction in total overnight food consumption associated with a marked decrease in the mass of the major feeding bout. After nine nights, however, cineole tolerance had developed as total food consumption had returned to the control amount. Compared to the control diet, the cineole diet was eaten in a larger number of smaller bouts, which were also eaten at a slower rate. The experiment was repeated with animals that had been accustomed to day-time feeding to take blood samples during feeding sessions. Feeding variables and blood concentration data for cineole were compared on the first and seventh day of the cineole diet. Although the total food consumed increased 2.5-fold after 7 days of the cineole diet, there was no increase in average blood cineole concentration, measured as the area under the concentration time curve. This indicates that induction of liver enzymes resulted in greater pre-systemic metabolism of cineole and reduced bioavailability. The maximum tolerated blood concentration of cineole also increased, suggesting some adaptation of the central nervous system to the cineole aversive effects. This appears to be the first report in a vertebrate herbivore that consumption of a dietary PSM leads to metabolism induction and that this contributes to development of tolerance to the PSM. Overall, herbivores adapt to newly encountered dietary PSMs by immediate changes in feeding behavior followed by development of increased metabolism of PSM and probably diminished cellular responsiveness to effects. PMID- 18401661 TI - Developmental and geographical variation in the chemical defense of the walkingstick insect Anisomorpha buprestoides. AB - Anisomorpha buprestoides, a walkingstick common in the southeastern United States, sprays chemicals that irritate and repel threatening insects, birds, or mammals. The active chemical in this substance was initially identified as a monoterpene dialdehyde. This compound can be present in several stereoisomeric forms, and subsequent studies have revealed that A. buprestoides produces at least three diastereomers: anisomorphal, dolichodial, and peruphasmal. However, no inquiry has been made to date into the geographical or developmental dependence of this variation. We report here that different populations of adult A. buprestoides spray either anisomorphal, or peruphasmal, or a mixture of the two stereoisomers. Additionally, offspring of a peruphasmal-producing population produced a variable mixture of anisomorphal and dolichodial but switched to peruphasmal upon reaching sexual maturity. This appears to be the first report of a developmentally regulated change in walkingstick insect chemical defense. Our results suggest a more complex role of these substances in the overall chemical ecology of walkingstick insects. PMID- 18401662 TI - Local therapy in stage IV breast cancer patients. PMID- 18401664 TI - Genomic alterations associated with early stages of breast tumor metastasis. AB - BACKGROUND: Molecular studies suggest that acquisition of metastatic potential occurs early in the development of breast cancer; mechanisms by which cells disseminate from the primary carcinomas and successfully colonize foreign tissues are, however, largely unknown. Thus, we examined levels and patterns of chromosomal alterations in primary breast tumors from node-negative (n = 114) and node-positive (n = 115) patients to determine whether specific genomic changes are associated with tumor metastasis. METHODS: Fifty-two genetic markers representing 26 chromosomal regions commonly altered in breast cancer were examined in laser microdissected tumor samples to assess levels and patterns of allelic imbalance (AI). Real time-PCR (RT-PCR) was performed to determine expression levels of candidate genes. Data was analyzed using exact unconditional and Student's t-tests with significance values of P < 0.05 and P < 0.002 used for the clinicopathological and genomic analyses, respectively. RESULTS: Overall levels of AI in primary breast tumors from node-negative (20.8%) and node positive (21.9%) patients did not differ significantly (P = 0.291). When data were examined by chromosomal region, only chromosome 8q24 showed significantly higher levels (P < 0.0005) of AI in node-positive primary tumors (23%) versus node-negative samples (6%). c-MYC showed significantly higher levels of gene expression in primary breast tumors from patients with lymph node metastasis. CONCLUSIONS: Higher frequencies of AI at chromosome 8q24 in patients with positive lymph nodes suggest that genetic changes in this region are important to the process of metastasis. Because overexpression of c-MYC has been associated with cellular dissemination as well as development of the premetastatic niche, alterations of the 8q24 region, including c-MYC, may be key determinants in the development of lymph node metastasis. PMID- 18401665 TI - A computational study of the development of epithelial acini: II. Necessary conditions for structure and lumen stability. AB - Simple epithelial tissues are organized as single layers of tightly packed cells that surround hollow lumens and form selective barriers separating different internal compartments of the body. The maintenance of epithelial structure and its function requires tight coordination and control of all the life processes of epithelial cells via cell-to-cell communication and signaling. These well balanced cellular systems are, however, quite often disturbed by genetic or environmental cues that may lead to the formation of epithelial tumors (carcinomas). In fact, more than a half of all diagnosed tumors are initiated from epithelial cells. It is, therefore, important to gain a greater understanding of the factors that form and maintain the epithelial structure, as well as the features of the acinar structure that are modified during cancer development as observable in experimental and clinical research. We address these questions using the bio-mechanical model of the developing hollow epithelial acini introduced in Rejniak and Anderson (Bull. Math. Biol. 70:677-712, 2008). Here, we propose several scenarios involving various bio-mechanical interactions between neighboring cells that result in abnormal acinar development. Whenever possible, we compare our computational results with known experimental cases of mutant acini. PMID- 18401666 TI - Isolation and characterization of mesenchymal stem cells derived from bone marrow of patients with Parkinson's disease. AB - Mesenchymal stem cells (MSCs) are capable of self-renewing and differentiating into multiple tissues; they are expected to become a source of cells for regenerative therapy. Compared to allogeneic MSCs, autologous MSCs from patients needing cell-based therapy may be an ideal alternative stem cell source. However, characterizations of MSCs from a disease state remains extremely limited. Therefore, we have isolated and characterized MSCs from Parkinson's disease (PD) patients and compared them with MSCs derived from normal adult bone marrow. Our results show that PD-derived MSCs are similar to normal MSCs in phenotype, morphology, and multidifferentiation capacity. Moreover, PD-derived MSCs are capable of differentiating into neurons in a specific medium with up to 30% having the characteristics of dopamine cells. At last, PD-derived MSCs could inhibit T-lymphocyte proliferation induced by mitogens. These findings indicate that MSCs derived from PD patients' bone marrow may be a promising cell type for cellular therapy and somatic gene therapy applications. PMID- 18401667 TI - Cytological properties of an Aedes albopictus mosquito cell line infected with Wolbachia strain wAlbB. AB - In vitro production of the obligate intracellular bacterium, Wolbachia pipientis, is essential to its manipulation as a genetic tool to spread transgenes within vector populations. We have adapted the Wolbachia-infected Aa23 Aedes albopictus mosquito cell line to Eagle's minimal medium, supplemented with nonessential amino acids, glutamine, and 20% fetal bovine serum. When plated at low densities, Aa23E cells grew as patchy monolayers, comprised of non-contiguous clusters of cells that gave rise to solid clumps of tightly adherent cells. Multicellular clumps eventually detached from the substrate and floated freely in the medium. Removal of Wolbachia by treatment with tetracycline did not alter the cytological properties of the host cells, which had a population doubling time of 4-5 d. The presence of Wolbachia was monitored by Giemsa staining of cytological preparations, polymerase chain reaction (PCR) amplification of Wolbachia 16S ribosomal DNA, and by simultaneous PCR amplification of ribosomal protein genes from Wolbachia and mosquito host cell genomes. Wolbachia morphology was pleomorphic, and Wolbachia DNA persisted in the culture medium for several weeks after degradation of PCR-amplifiable host cell DNA. PMID- 18401668 TI - Metabolic syndrome after laparoscopic bariatric surgery. AB - BACKGROUND: Metabolic syndrome (MS) is common among morbidly obese patients undergoing bariatric surgery. The aim of this study was to assess the impact and predictors of bariatric surgery on the resolution of MS. METHODS: Subjects included 286 patients [age 44.0 +/- 11.5, female 78.2%, BMI 48.7 +/- 9.4, waist circumference 139 +/- 20 cm, AST 23.5 +/- 14.9, ALT 30.0 +/- 20.1, type 2 diabetes mellitus (DM) 30.1% and MS 39.2%] who underwent bariatric surgery. RESULTS: Of the entire cohort, 27.3% underwent malabsorptive surgery, 55.9% underwent restrictive surgery, and 16.8% had combination restrictive malabsorptive surgery. Mean weight loss was 33.7 +/- 20.1 kg after restrictive surgery (follow up period 298 +/- 271 days), 39.4 +/- 22.9 kg after malabsorptive surgery (follow-up period 306 +/- 290 days), and 28.3 +/- 14.1 kg after combination surgery (follow-up period 281 +/- 239 days). Regardless of the type of bariatric surgery, significant improvements were noted in MS (p values from <0.0001-0.01) as well as its components such as DM (p values from <0.0001 0.0005), waist circumference (p values <0.0001), BMI (p values <0.0001), fasting serum triglycerides (p values <0.0001 to 0.001), and fasting serum glucose (p values <0.0001). Additionally, a significant improvement in AST/ALT ratio (p value = 0.0002) was noted in those undergoing restrictive surgery. Multivariate analysis showed that patients who underwent malabsorptive bariatric procedures experienced a significantly greater percent excess weight loss than patients who underwent restrictive procedures (p value = 0.0451). Percent excess weight loss increased with longer postoperative follow-up (p value <0.0001). CONCLUSIONS: Weight loss after bariatric surgery is associated with a significant improvement in MS and other metabolic factors. PMID- 18401669 TI - What is the quality of information about bariatric surgery on the internet? AB - BACKGROUND: The purpose of this study was to evaluate the role of Internet on patients scheduled for bariatric procedures and the quality of information available on different websites. METHODS: Between July 2003 to July 2005, patients undergoing bariatric surgical procedures completed a survey. Data were collected prospectively. One hundred valid surveys were returned. Independently, two bariatric surgeons evaluated available French and English websites using major search engines. RESULTS: Forty-two of 100 patients (42%) sought information about bariatric surgery on the Internet. Seventy-four percent of these patients (n = 31/42) used search engines with 81% visiting less than ten websites. According to the patient's evaluation, 58% of the websites visited did not provide technical details of any surgical bariatric procedures, and only 61% provided information regarding postoperative weight loss. Furthermore, 58% of websites did not provide information about the laparoscopic approach, and 54% did not give any information on potential postoperative complications. Bariatric surgeon's evaluation was similar except for two differences: laparoscopic approach and postoperative weight loss information were discussed in 90% (p < 0.001) and 43% (p < 0.1) of visited websites, respectively. CONCLUSION: When the Internet was used to search for information about bariatric surgery, search engines were preferentially used but search duration was short. Available Internet websites can be considered as moderately reliable; however, 25% of visited websites contain misleading information. Comparison between patients and surgeons views showed that patients were effective in detecting misleading information. PMID- 18401670 TI - The effect of laparoscopic Roux-en-Y gastric bypass on fibromyalgia. AB - BACKGROUND: Fibromyalgia is a chronic debilitating disorder affecting 3-5% of the US population. Treatment of this disorder is a challenge. The incidental finding of improvement of fibromyalgia following laparoscopic Roux-en-Y gastric bypass stimulated us to study this phenomenon. METHODS: A retrospective chart review of patients with fibromyalgia who underwent laparoscopic Roux-en-Y gastric bypass. RESULTS: Postoperative decrease in median of BMI from 49.4 to 29.7 was significant (p value = 0.0010). This was associated with statistically significant improvement in median of pain score (p value = 0.0010) and median points of tenderness (p value = 0.0010). CONCLUSION: Significant weight loss following laparoscopic Roux-en-Y gastric bypass is associated with resolution or improvement of fibromyalgia. Consequently, the bariatric surgeon should be a member of the multidisciplinary team approach for treating fibromyalgia. PMID- 18401672 TI - Sleeve gastrectomy in a 10-year-old child. AB - Adolescents bariatric surgery (ABS) in morbid obesity (MO), with or without comorbid conditions, is and will be more and more indicated. Restrictive operations have the advantage of no influence on absorption. Laparoscopic sleeve gastrectomy (LSG) can be an excellent alternative. A LSG was done in a 10-year old boy, body mass index (BMI) 42, who has Blount's disease (tibia vara) with severe pain at the knee joints that made him a wheelchair-bound person. He had a LSG and gallbladder removal without incidents. Eight months later, he has a BMI 28 and almost all his knees pain is gone. No side effects have been detected. A LSG may be the ideal bariatric operation for ABS with MO. PMID- 18401671 TI - Influence of morbid obesity and insulin resistance on gene expression levels of AQP7 in visceral adipose tissue and AQP9 in liver. AB - BACKGROUND: Glycerol production and its efflux from adipocytes to the liver are key to modulate lipid and glucose homeostasis. Aquaporin 7 (AQP7) is an aquaglyceroporin that acts as the adipose glycerol channel, whereas aquaporin 9 (AQP9) is the specific channel operating in the liver. The aim of the present work was to evaluate the effect of obesity and type 2 diabetes mellitus (T2DM) on gene expression levels of AQP7 in visceral adipose tissue (VAT) and AQP9 in liver. METHODS: VAT and liver biopsies obtained from 20 women were used in the study. Patients were classified as lean or obese with the last group being further subclassified as normoglycemic (NG), patients with impaired glucose tolerance (IGT), or with T2DM. Anthropometric measurements as well as circulating metabolites, hormones, and adipokines were determined. Real-time polymerase chain reaction analyses were performed to quantify transcript levels of AQP7 in VAT and AQP9 in the liver. RESULTS: Gene expression levels of AQP7 in VAT showed a tendency toward an increase (P = 0.065) in obese patients (both NG and T2DM) compared to lean subjects. AQP9 showed a significant downregulation in the hepatic biopsies obtained from obese T2DM patients compared to obese NG and IGT patients (P = 0.028). CONCLUSION: The tendency toward an elevation of mRNA expression of VAT AQP7 in obesity together with the decreased hepatic AQP9 expression observed in obese T2DM subjects suggests a potential role in facilitating glycerol release from adipose tissue and reducing glycerol entry into hepatocytes in obesity and T2DM, respectively. PMID- 18401673 TI - Tailoring a fruit and vegetable intervention on novel motivational constructs: results of a randomized study. AB - BACKGROUND: Tailored health communications to date have been based on a rather narrow set of theoretical constructs. PURPOSE: This study was designed to test whether tailoring a print-based fruit and vegetable (F & V) intervention on relatively novel constructs from self-determination theory (SDT) and motivational interviewing (MI) increases intervention impact, perceived relevance, and program satisfaction. The study also aimed to explore possible user characteristics that may moderate intervention response. METHODS: African American adults were recruited from two integrated health care delivery systems, one based in the Detroit Metro area and the other in the Atlanta Metro area, and then randomized to receive three tailored newsletters over 3 months. One set of newsletters was tailored only on demographic and social cognitive variables (control condition), whereas the other (experimental condition) was tailored on SDT and MI principles and strategies. The primary focus of the newsletters and the primary outcome for the study was fruit and vegetable intake assessed with two brief self-report measures. Preference for autonomy support was assessed at baseline with a single item: "In general, when it comes to my health I would rather an expert just tell me what I should do". Most between-group differences were examined using change scores. RESULTS: A total of 512 (31%) eligible participants, of 1,650 invited, were enrolled, of which 423 provided complete 3-month follow-up data. Considering the entire sample, there were no significant between-group differences in daily F & V intake at 3 month follow-up. Both groups showed similar increases of around one serving per day of F & V on the short form and half a serving per day on the long form. There were, however, significant interactions of intervention group with preference for autonomy-supportive communication as well as with age. Specifically, individuals in the experimental intervention who, at baseline, preferred an autonomy-supportive style of communication increased their F & V intake by 1.07 servings compared to 0.43 servings among controls. Among younger controls, there was a larger change in F & V intake, 0.59 servings, than their experimental group counterparts, 0.29 servings. Conversely, older experimental group participants showed a larger change in F & V, 1.09 servings, than older controls, 0.48. CONCLUSION: Our study confirms the importance of assessing individual differences as potential moderators of tailored health interventions. For those who prefer an autonomy-supportive style of communication, tailoring on values and other motivational constructs can enhance message impact and perceived relevance. PMID- 18401674 TI - Immunological function of thymoma and pathogenesis of paraneoplastic myasthenia gravis. AB - Thymoma and thymic carcinoma are the representative tumors arising from the thymic epithelium. Thymoma is well known for association with autoimmune diseases including myasthenia gravis, suggesting its biological activity. Herein, recent progress in research of thymoma is reviewed with reference to its immunological function. Myasthenia gravis is frequently associated with WHO type B1 and B2 thymomas. These types of thymomas hold a significant number of CD4(+)CD8(+) double-positive T cells, and at the same time, the neoplastic epithelial cells express HLA-DR molecules at a slightly reduced level compared with the normal thymus. The impaired expression of HLA-DR molecules in neoplastic epithelial cells of thymomas possibly affects positive selection of CD4(+)CD8(-) single positive T cells and may result in alteration of its repertoire. The function of thymoma neoplastic cells as the cortical epithelium of the thymus and the morphological resemblance of thymomas to the cortex suggest that thymoma is of cortical epithelial origin; this might imply that thymoma lacks the functional medulla where professional antigen-presenting cells are engaged in negative selection. These findings suggest that thymoma generates autoreactive T cells causing autoimmunity. Further investigation on immunological function of thymoma is supposed to elucidate the pathogenesis of thymoma-related autoimmunity and the high affinity of thymoma with myasthenia gravis. In addition, studying the biology of thymoma is also expected to contribute to further understanding of T cell development and immunological tolerance in the human, because thymoma can be considered an acquired thymus. PMID- 18401675 TI - Radical sublobar resection for lung cancer. AB - Sublobar resection (segmentectomy or wedge resection) has not historically been the strategy of choice for treating lung cancer and is performed only as a compromise for patients with early-stage lung cancer accompanied by poor lung function. Although some studies have advocated higher rates of local recurrence and a poorer prognosis after sublobar resection than after lobectomy, most others have found promising outcomes after sublobar resection in selected patients. Yet even now, when early-stage small lung cancers are being detected with increasing frequency, sublobar resection has yet to become the treatment of choice. This review summarizes surgical outcomes of sublobar resection compared with lobectomy in the literature. Current evidence indicates that radical sublobar resection should be considered as an alternative for cT1N0 lung cancer of < or = 2 cm, even in low-risk patients. A foundation is thus established for starting a new series of randomized controlled trials, which could bring about revolutionary changes to surgery for lung cancer in the era of early detection. PMID- 18401676 TI - Bilateral pulmonary artery banding for hypoplastic left heart syndrome and related anomalies. AB - OBJECTIVE: Bilateral pulmonary artery banding is considered an option for initial palliation in high-risk patients with hypoplastic left heart syndrome or related anomalies. However, there are potential interim morbidities, including ductal constriction, compromised growth of the ascending aorta, and pulmonary artery stenosis at the banding site. In addition, there are still controversies regarding the optimal timing and choice of the second-stage operation. METHODS: Between February 2003 and October 2005, twelve high-risk patients with hypoplastic left heart syndrome or related anomalies underwent bilateral pulmonary artery banding. An atrial septal defect was created simultaneously when the septum was intact or the defect was restrictive. After the procedure, intravenous prostaglandin E(1) was continuously administered. RESULTS: The median weight was 3.0 kg (range 1.8-3.6 kg), and the median age was 16 days (range 0-27 days). Atrial septum defect creation was performed simultaneously in two patients. Ten of the twelve patients were discharged from the hospital. One patient died of progressive multiple organ failure and another patient died of cerebral hemorrhage. Eight patients developed various degrees of ductal constriction and required dose adjustment of prostaglandin E(1). Two patients underwent ductal arteriosus stent placement: one was unsuccessful owing to the size mismatch between the stent and the ductus. The diameter of the ascending aorta remained unchanged until the time of the second-stage operation, except in two patients who had a sufficiently sized aorta at the time of pulmonary artery banding. Pulmonary arterial branch stenosis developed at the banding site in three patients. At the time of the second-stage operation, the Norwood procedure was performed in seven patients, biventricular repair in two, and Norwood plus Glenn procedure in one. CONCLUSION: Bilateral pulmonary artery banding is an option to salvage high-risk neonates with hypoplastic left heart syndrome or related anomalies. However, it is associated with high interstage morbidity, and patients may benefit from early conversion to the Norwood operation. PMID- 18401677 TI - Pharmacokinetic analysis of flomoxef in children undergoing cardiopulmonary bypass and modified ultrafiltration. AB - OBJECTIVE: Cardiopulmonary bypass (CPB) induces changes in the pharmacokinetics of drugs. The purpose of this study was to model the pharmacokinetics of flomoxef, a cephalosporin antibiotic, in pediatric cardiac surgery. METHODS: Each patient received a flomoxef dose of 30 mg/kg as a bolus after the induction of anesthesia and an additional dose (1 g for a child weighing < 10 kg, 2 g for > or = 10 kg) was injected into the CPB prime. Modified ultrafiltration (MUF) was routinely performed. Blood samples, urine, and ultrafiltrate were collected. In seven patients (group I), serum flomoxef concentration-time courses were analyzed by a modified two-compartment model. Utilizing the estimated parameters, serum concentrations were simulated in another eight patients (group II). RESULTS: The initiation of CPB resulted in an abrupt increase in serum flomoxef concentrations in group I; however, concentrations declined biexponentially. The amount of excreted flomoxef in the urine and by MUF was 47% +/- 8% of the total administered dose. In group II, an excellent fit was found between the values calculated by the program and the observed serum concentrations expressed; most of the performance errors were <1.0. There was no difference in any kinetic parameter between group I and groups I + II (n = 15). CONCLUSION: The pharmacokinetics of flomoxef in children undergoing CPB and MUF were well fitted to a modified two-compartment model. Using the kinetic data from this study, the individualization of dosage regimens for prophylactic use of flomoxef might be possible. PMID- 18401678 TI - Role of oxidative stress in hypertrophied myoblasts stimulated by isoproterenol. AB - OBJECTIVE: Hydrogen peroxide (H(2)O(2)), one of the oxyradical members, has been shown to have insulin-like effects, and endogenous H(2)O(2) may have the ability to improve impaired glucose metabolism. As a mechanism contributing to progression of hypertrophy, we hypothesized that the formation of H(2)O(2) to improve glucose uptake could be an oxidative stress resulting in apoptotic cell death. METHODS: We used cultured myoblasts (H9c2) stimulated by isoproterenol as a model to focus on glucose metabolism, oxidative stress, apoptosis, and extracellular signal-regulated protein kinases (ERKs) expression. RESULTS: In hypertrophied myoblasts, anaerobic glycolysis became prominent at day 7 of isoproterenol stimulation (ISO7; isoproterenol was administered for 7 days). Hydroperoxides production started to increase at day 5 of isoproterenol stimulation (ISO5) and peaked at ISO7. Apoptotic cell death was significantly increased in ISO5 and ISO7. Temporary ERKs suppression was also found at ISO5 and ISO7. Change in glucose metabolism led to stimulation of hydroperoxides production, apoptotic cell death and suppression of ERKs. CONCLUSION: We concluded that oxidative stress was increased during the progression of hypertrophied myoblasts in association with impaired glucose metabolism and increased apoptotic cell death. Suppression of ERKs may be involved in signal transduction of apoptotic cell death. PMID- 18401679 TI - Communication between the right pulmonary artery and left atrium with aneurysm formation. AB - Communication between the pulmonary artery and the left atrium is a rare congenital anomaly. A 7-year-old boy was referred to our hospital with a diagnosis of pulmonary arteriovenous fistula. Referral to a pediatric cardiologist had been delayed because he had only slight cyanosis and no heart murmur. Catheterization and computed tomography scans confirmed that the right pulmonary branches and pulmonary venous returns were normal and that an additional aneurysmal communication was located between the right pulmonary artery and the left atrium. Surgery proceeded uneventfully together with a cardiopulmonary bypass. The connection from the right pulmonary artery to the aneurysm was ligated, and the connection from the left atrium to the aneurysm was closed with a polytetrafluoroethylene patch. We describe this rare anomaly with cyanosis and no heart murmur. PMID- 18401680 TI - Extraskeletal osteosarcoma of the pleura: a case report. AB - We herein present an exceedingly rare case of pleural osteosarcoma that was surgically resected in an elderly patient. A 74-year-old man, complaining of a dry cough and breathlessness on effort, was found to have massive pleural effusion on a chest X-ray. The chest CT and MRI scans indicated a massive effusion and a pleural tumor encasing the left lower lung. The resected tumor was histologically an osteosarcoma, measuring 11.3 x 9.0 x 6.0 cm. Because there was no evidence of any other primary tumor, the diagnosis of extraskeletal osteosarcoma was appropriate. PMID- 18401681 TI - Extensive total arch replacement via clamshell incision in a patient with aortic arch aneurysm and Stanford type B aortic dissection. AB - The utility of the clamshell approach, namely, a bilateral anterior thoracotomy incision for treating extensive thoracic aortic disease was described by Kouchoukos et al. in 2001 and by Doss et al. in 2003. We describe the utility of this approach for treating aortic arch aneurysm with Stanford type B aortic dissection. PMID- 18401682 TI - Video-assisted thoracoscopic surgery for pulmonary arteriovenous malformations: report of five cases. AB - We experienced five cases of pulmonary arteriovenous malformations (PAVMs) that were successfully treated by video-assisted thoracoscopic surgery. Four malformations were treated by local wedge resection and one was treated by segmentectomy. Criteria for patient selection for surgery were peripheral and solitary lesions, with feeding arteries larger than 3 mm. Postoperative hospital stays were 1-7 days (median, 2 days). All patients showed unchanged or increased values of PaO(2) in arterial blood after operation. No major postoperative complication occurred in any patient, but a persistent air leak for 5 days occurred in the one patient who was treated by segmentectomy. No growth of accessory vessels or untreated malformations were seen in any patient throughout the follow-up period of 14-54 months. Thoracoscopic surgical resection for well selected patients provides a high certainty of eliminating fistulae and was associated with lower morbidity, lower mortality, and shorter hospital stays. PMID- 18401683 TI - Inhaled foreign body overlooked for 25 years in an adult. AB - A 30-year-old man was admitted to our hospital because of refractory pneumonia. He had been repeatedly hospitalized owing to pneumonia recurring in the left lower lung field and had a 7-day history of hemoptysis at the age of 11. Chest computed tomography demonstrated reduced volume of the left lower lobe with parenchymal consolidation. The patient preferred surgery as first-line treatment rather than other interventions because he had suffered from refractory pneumonia or local pleuritis for a long time. Left lower lobectomy was performed. The resected specimen revealed obstruction of the posterior basal segment bronchus in which a foreign body (FB) was found embedded in granulation tissue. The FB was a plastic air-pistol pellet. After further questioning, the patient remembered aspirating the pellet while playing with the pistol at the age of 5. When it is difficult to remove a long-standing intrabronchial FB, we recommend surgery. PMID- 18401684 TI - Primary pulmonary mucinous (colloid) adenocarcinoma. AB - We report a rare case of primary pulmonary mucinous (colloid) adenocarcinoma (MC) in a 79-year-old man. A computed tomographic scan of the chest showed a round, well-defined nodule in his right upper lobe that enlarged slowly over 12 months from 1.7 cm to 2.0 cm. Wedge resection of the right upper lobe was performed to obtain a definitive diagnosis by video-assisted thoracoscopic surgery. Macroscopically, the cut surface showed that the nodule was well demarcated and filled with a yellowish-white gelatinous substance. The postoperative histological diagnosis was primary pulmonary MC. PMID- 18401685 TI - Tracheal chondrosarcoma. AB - Tracheal chondrosarcomas are rare, with only 14 cases previously documented. We report a case of chondrosarcoma of the trachea and review other published cases. Our patient was a 34-year-old man who began to feel shortness of breath 8 months after initial hemoptysis. Transverse and coronal computed tomography demonstrated a tumor that was obstructing nearly 80% of the tracheal lumen at the thoracic inlet. A bronchoscopic biopsy specimen led to an initial diagnosis of chondroma. Five tracheal rings from the third to the seventh including the tumor were completely resected, and an end-to-end anastomosis was performed. The histological diagnosis of the surgical specimen showed it to be a chondrosarcoma. The patient was alive without recurrence at 6 years 4 months after surgery. Because recurrence after incomplete resection and malignant transformation can occur with chondromas, radical resection of the trachea is recommended for all cartilaginous tumors of the trachea. PMID- 18401686 TI - Biofilm formation and partial biodegradation of polystyrene by the actinomycete Rhodococcus ruber: biodegradation of polystyrene. AB - Polystyrene, which is one of the most utilized thermoplastics, is highly durable and is considered to be non-biodegradable. Hence, polystyrene waste accumulates in the environment posing an increasing ecological threat. In a previous study we have isolated a biofilm-producing strain (C208) of the actinomycete Rhodococcus ruber that degraded polyethylene films. Formation of biofilm, by C208, improved the biodegradation of polyethylene. Consequently, the present study aimed at monitoring the kinetics of biofilm formation by C208 on polystyrene, determining the physiological activity of the biofilm and analyzing its capacity to degrade polystyrene. Quantification of the biofilm biomass was performed using a modified crystal violet (CV) staining or by monitoring the protein content in the biofilm. When cultured on polystyrene flakes, most of the bacterial cells adhered to the polystyrene surface within few hours, forming a biofilm. The growth of the on polystyrene showed a pattern similar to that of a planktonic culture. Furthermore, the respiration rate, of the biofilm, exhibited a pattern similar to that of the biofilm growth. In contrast, the respiration activity of the planktonic population showed a constant decline with time. Addition of mineral oil (0.005% w/v), but not non-ionic surfactants, increased the biofilm biomass. Extended incubation of the biofilm for up to 8 weeks resulted in a small reduction in the polystyrene weight (0.8% of gravimetric weight loss). This study demonstrates the high affinity of C208 to polystyrene which lead to biofilm formation and, presumably, induced partial biodegradation. PMID- 18401687 TI - Fe(III), Cr(VI), and Fe(III) mediated Cr(VI) reduction in alkaline media using a Halomonas isolate from Soap Lake, Washington. AB - Hexavalent chromium is one of the most widely distributed environmental contaminants. Given the carcinogenic and mutagenic consequences of Cr(VI) exposure, the release of Cr(VI) into the environment has long been a major concern. While many reports of microbial Cr(VI) reduction are in circulation, very few have demonstrated Cr(VI) reduction under alkaline conditions. Since Cr(VI) exhibits higher mobility in alkaline soils relative to pH neutral soils, and since Cr contamination of alkaline soils is associated with a number of industrial activities, microbial Cr(VI) reduction under alkaline conditions requires attention.Soda lakes are the most stable alkaline environments on earth, and contain a wide diversity of alkaliphilic organisms. In this study, a bacterial isolate belonging to the Halomonas genus was obtained from Soap Lake, a chemically stratified alkaline lake located in central Washington State. The ability of this isolate to reduce Cr(VI) and Fe(III) was assessed under alkaline (pH = 9), anoxic, non-growth conditions with acetate as an electron donor. Metal reduction rates were quantified using Monod kinetics. In addition, Cr(VI) reduction experiments were carried out in the presence of Fe(III) to evaluate the possible enhancement of Cr(VI) reduction rates through electron shuttling mechanisms. While Fe(III) reduction rates were slow compared to previously reported rates, Cr(VI) reduction rates fell within range of previously reported rates. PMID- 18401688 TI - Larvae of chigger mites Neotrombicula spp. (Acari: Trombiculidae) exhibited Borrelia but no Anaplasma infections: a field study including birds from the Czech Carpathians as hosts of chiggers. AB - Chigger mites were collected from 1,080 wild birds of 37 species at Certak (Czech Republic), in the western Carpathian Mountains, from 29 July to 24 September 2005. The prevalence of infestation with chigger larvae was 7%. A total of 325 chigger specimens from 10 bird species was identified and three chigger species were found: Neotrombicula autumnalis, N. carpathica, and N. inopinata, the latter two species being reported on new hosts. Neotrombicula carpathica is reported in the Czech Republic for the first time. A total of 509 chigger larvae found on 79 host specimens were examined by polymerase chain reaction (PCR) for the presence of Borrelia burgdorferi s.l. DNA (fragments of the rrf (5S)--rrl (23S) intergenic spacer), and Anaplasma phagocytophilum DNA (epank1 gene). A fragment of specific Borrelia DNA was amplified through PCR in one sample, and the PCR product was further analyzed by reverse line blotting assay, whereby both genospecies of B. garinii and B. valaisiana were proved. This sample pooled five chigger larvae collected from one Sylvia atricapilla on 11 August 2005. No A. phagocytophilum DNA was amplified. We conclude that larvae of the genus Neotrombicula can be infected with Borrelia genospecies originated from their present or former hosts. PMID- 18401689 TI - Impact of preoperative antiandrogen medication and nerve-sparing surgery on the outcome of radical prostatectomy. AB - OBJECTIVES: Radical prostatectomy in antiandrogen-medicated patients, as well as nerve-sparing surgery, has increased within recent years. We investigated the impact of antiandrogen medication on loss of blood and of nerve-sparing surgery on continence and potency. METHODS: A total of 401 men who underwent radical prostatectomy between January 1995 and September 1999 at our clinic were asked about antiandrogenic medication prior to radical prostatectomy and about daily life activities, overall satisfaction, voiding ability, and sexual activities. Consumption of erythrocyte concentrates and cryoprecipitated plasma was taken from the records. RESULTS: The average follow-up was 36 months. Seventy-two percent (n = 289) of our patients replied. Sixty-seven men (23%) who replied to our questionnaire had received antiandrogen medication prior to radical prostatectomy. In 53 (18%) of the men, the nerve-sparing technique was used. In antiandrogen-medicated patients, we observed a significantly elevated consumption of erythrocyte concentrates (antiandrogen-medicated 0.93, not antiandrogen medicated 0.44; P = 0.013) and of cryoprecipitated plasma (antiandrogen-medicated 0.39, not antiandrogen-medicated 0.08; P = 0.010). Patients who underwent the nerve-sparing technique reported better results in daily life activities, general health status, International Prostate Symptom Score (IPSS), and continence. CONCLUSION: Our results point to a higher quality of life in patients who had undergone nerve-sparing surgery. PMID- 18401690 TI - Psychosis and the experience of employment. AB - INTRODUCTION: Although unemployment is common among those diagnosed with psychotic illnesses, few studies have looked directly at the experience of those who have gained competitive paid employment subsequent to being diagnosed. This study explored the experiences of people diagnosed with schizophrenia or schizoaffective disorder in relation to paid employment. METHODS Eight participants with experience of paid employment were interviewed. The data were analysed using Interpretative Phenomenological Analysis (IPA) [Smith et al. (In: Murray, Chamberlain, editors) Qualitative health psychology, 1999]. RESULTS: Three super-ordinate themes; 'Coping', 'Interpersonal support and reactions', and 'Personal change' were identified. CONCLUSIONS: Interpersonal support seemed to be lacking in workplaces, where discrimination was a reality for people with psychosis. Further study into what facilitates coping with symptoms in the workplace is needed. PMID- 18401691 TI - Brief report: attention effect on a measure of social perception. AB - A measure of social perception (CASP) was used to assess differences in social perception among typically developing children, children with autistic spectrum disorders (ASD), and children with Attention Deficit/Hyperactivity Disorder (ADHD). Significant between-group differences were found in recognition of emotions in video, with children diagnosed with ADHD or ASD performing more poorly than controls on measures of knowledge of nonverbal cues and emotional expression. The number of inattention symptoms was found to be a significant contributor to poorer video interpretation across diagnostic groups. An effect of attention on poor social perception was found that may be unique to the ASD group. Clinicians may overestimate the social deficits of children with ASD if they also have symptoms of inattention, while researchers may need to control for the effects of inattention in their studies. PMID- 18401692 TI - The Classroom Observation Schedule to Measure Intentional Communication (COSMIC): an observational measure of the intentional communication of children with autism in an unstructured classroom setting. AB - The Classroom Observation Schedule to Measure Intentional Communication (COSMIC) was devised to provide ecologically valid outcome measures for a communication focused intervention trial. Ninety-one children with autism spectrum disorder aged 6 years 10 months (SD 16 months) were videoed during their everyday snack, teaching and free play activities. Inter-rater reliability was high and relevant items showed significant associations with comparable items from concurrent Autism Diagnostic Observation Schedule-Generic (Lord et al. 2000, J Autism Dev Disord 30(3):205-223) assessments. In a subsample of 28 children initial differences in rates of initiations, initiated speech/vocalisation and commenting were predictive of language and communication competence 15 months later. Results suggest that the use of observational measures of intentional communication in natural settings is a valuable assessment strategy for research and clinical practice. PMID- 18401693 TI - The effectiveness of parent-child interaction therapy for families of children on the autism spectrum. AB - We report the results of a pilot trial of an evidence-based treatment-Parent Child Interaction Therapy (PCIT; Eyberg et al. Psychopharmacology Bulletin, 31(1), 83-91, 1995) for boys aged 5-12 with high functioning autism spectrum disorders and clinically significant behavioral problems. The study also included an investigation of the role of shared positive affect during the course of therapy on child and parent outcomes. The intervention group showed reductions in parent perceptions of child problem behaviors and child atypicality, as well as an increase in child adaptability. Shared positive affect in parent child dyads and parent positive affect increased between the initial and final phases of the therapy. Parent positive affect after the first phase was related to perceptions of improvement in problem behaviors and adaptive functioning. PMID- 18401694 TI - When prototypes are not best: judgments made by children with autism. AB - The current study used a factorial comparison experimental design to investigate conflicting findings on prototype effects shown by children with autism (Klinger and Dawson, Dev Psychopathol 13:111-124, 2001; Molesworth et al., J Child Psychol Psychiatry 46:661-672, 2005). The aim was to see whether children with high functioning autism could demonstrate prototype effects via categorization responses and whether failure to do so was related to difficulty understanding ambiguous task demands. Two thirds of the autism group did show an effect. The remainder, a sub-group defined by performance on a control task, did not. The discussion focuses on the influence of heterogeneity within the autism group and the ability to resolve ambiguity on task performance. Finally, an alternative experimental design is recommended for further research into these issues. PMID- 18401695 TI - Complete genome sequence of attenuated low-temperature Thiverval strain of classical swine fever virus. AB - The Thiverval vaccine strain of classical swine fever virus (CSFV) was derived from virulent Alfort strain through the serial passages in cells at 29-30 degrees C. In this study, we determined the complete genome sequence of this strain and found that its genome contains one open reading frame (ORF) that encodes a polyprotein with 3,898 amino acids. The 5'-UTR of Thiverval is 373 nt long with only one mutation at position 220. In contrast, the length of 3'-UTR is highly heterogeneous ranging from 233 to 259 bp. The heterogeneity of length of the 3' UTR was due to an insertion of a variable length of T-rich sequence ranging from 6 to 32 nt. The insertion may change the structure and free energy of the 3'-UTR, resulting in a destabilization of the 3'-UTR. Sequence alignment of Thiverval and other CSFV strains showed 85.2-99.6% identities at the nucleotide level and 92.5 99.5% at the amino acid level. The phylogenetic tree analysis of the complete ORF, partial region of E2, and NS5B suggests that the CSFV Thiverval strain belongs to genetic group 1 and subgroup 1.1. The results from this study provide insight into the molecular mechanism of the attenuation of Thiverval vaccine strain. PMID- 18401696 TI - Swine infection with H9N2 influenza viruses in China in 2004. AB - In 2004, H9N2 influenza A viruses were isolated from pigs with respiratory syndrome in commercial swine farms in Henan province, China. Antigenic and genetic characterization were performed for seven swine H9N2 influenza viruses. The hemagglutinin antigenicity of swine H9N2 viruses was similar to those of avian H9N2 viruses of A/duck/Hong Kong/Y280/1997 (Dk/HK/Y280/97)-like sublineage prevalent in China. It is noteworthy that the neuraminidase of these isolates had no deletions in the stalk, which was seldom observed in those viruses of Dk/HK/Y280/97-like sublineage. Genetic analysis revealed that all seven isolates had an -R-S-S-R- motif at the HA cleavage site, which was the same as those of Dk/HK/Y280/97-like viruses established in avian population in China. Phylogenetic analyses showed that the seven swine H9N2 viruses were completely derived from avian influenza viruses of Dk/HK/Y280/97-like sublineage. The present results indicated that avian-to-pig interspecies transmission of H9N2 viruses continued to exist in China through 2004; therefore, surveillance of swine influenza should be given a high priority. PMID- 18401697 TI - Genome characterization and genetic diversity of beet curly top Iran virus: a geminivirus with a novel nonanucleotide. AB - Beet curly top Iran virus (BCTIV) was previously reported as a distinct curtovirus in Iran. Complete nucleotide sequences of three BCTIV isolates, one each from central, southern, and south eastern Iran were determined to be 2844, 2844, and 2845 nt long, respectively. BCTIV shared highest nucleotide sequence identity (52.3%) with Spinach curly top virus (SpCTV) and lowest identity (46.6%) with Horseradish curly top virus (HrCTV). The BCTIV genome comprises three virion sense (V1, V2, and V3) and two complementary-sense (C1 and C2) ORFs. ORFs C3 and C4 were not found in BCTIV genome. Based on a comparison of nucleotide sequence identity of individual genes, the three virion-sense ORFs were 72.7-79.9% related to the corresponding ORFs of curtoviruses, whereas no significant relationship was found between the C1 and C2 ORFs of BCTIV and curtoviruses. These two ORFs, however, were only distantly related with those of mastreviruses. Similar to the latter viruses, the BCTIV genome comprises two intergenic regions. The BCTIV large intergenic region included a sequence capable of forming a stem loop structure and a novel nonanucleotide (TAAGATT/CC) with a unique nick site. Phylogenetic analysis using deduced amino acid sequence of individual ORFs revealed that the V2 and V3 ORFs are monophyletic and the V1 ORF is classified with the related ORF of curtoviruses. Whereas the two complementary-sense ORFs are grouped with those of mastreviruses. Computer-based prediction suggested that BCTIV has a chimeric genome which may have arisen by a recombination event involving curto- and mastrevirus ancestors. Percent nucleotide sequence identities of the coat protein gene of ten isolates of BCTIV, collected from a wide range of geographical regions in Iran, varied from 87.1 to 99.9, with the isolates being distributed between two subgroups. Based on biological and molecular properties, BCTIV is proposed as a new member of the genus Curtovirus. PMID- 18401698 TI - Correlates of any condom use among Russian narcology patients reporting recent unprotected sex. AB - The purpose of this study was to assess whether HIV/sexually transmitted infection (STI) risk factors: risky sex (multiple sex partners and sex trade involvement), past HIV or STI diagnosis and substance use (at risk drinking and injection drug use) are associated with the outcome any condom use in the past 6 months among Russian narcology hospital patients. Participants (N = 178) included only those who reported unprotected sex in the past 6 months and were aged 18-55 years and 76% male. Any condom use in the past 6 months was reported by 55% of the sample. History of STIs was reported by 43% of participants; 15% were HIV infected. Regression analyses adjusted for demographics demonstrated that those reporting multiple sex partners (OR(adj) = 4.2, 95% CI = 2.0-8.7) and sex trade involvement (OR(adj) = 2.4, 95% CI = 1.1-5.1) in the past 6 months had significantly higher odds of reporting any condom use in this same timeframe. HIV/STI and substance use were not associated with increased odds of condom use. PMID- 18401699 TI - Adherence to both cotrimoxazole and placebo is associated with improved survival among HIV-infected Zambian children. AB - In the CHAP randomized placebo-controlled trial of cotrimoxazole prophylaxis in HIV-infected Zambian children conducted between 2001 and 2003, cotrimoxazole was associated with significant mortality reductions. In a secondary analysis we used Cox regression models to estimate the association between adherence measured by bottle weights and caregiver report and subsequent mortality in children surviving >28 days (n = 496, 153 deaths). Adherence was high and similar in both cotrimoxazole and placebo groups; adherence from bottle weights was 100% at 71% of visits, while caregivers reported 100% adherence at 79% of visits. Every 10% lower adherence to cotrimoxazole or placebo measured by bottle weights was associated with a 10-11% increase in mortality risk. Effects remained after adjustment for baseline predictors of survival and for current and recent change in primary caregiver. Caregiver-reported adherence was not associated with survival. The association between bottle-weight adherence to placebo and survival is likely capturing unmeasured caregiver effects, whose identification will be essential for quantifying the impact of antiretroviral therapy (ART) adherence on clinical outcomes in children. PMID- 18401700 TI - A population-based survey of the prevalence of HIV, syphilis, hepatitis B and hepatitis C infections, and associated risk factors among young women in Vitoria, Brazil. AB - OBJECTIVE: To estimate the prevalence of HIV, hepatitis B (HBV) and C (HCV), and syphilis infections and associated risk exposures in a population-based sample of young women in Vitoria, Brazil. METHODS: From March to December 2006, a cross sectional sample of women aged 18-29 years was recruited into a single stage, population-based study. Serological markers of HIV, HBV, HCV, and syphilis infections and associated risk exposures were assessed. RESULTS: Of 1,200 eligible women, 1,029 (85.8%) enrolled. Median age was 23 (interquartile range 20 26) years; 32.2% had < or = 8 years of education. The survey weighted prevalence estimates were: HIV, 0.6% [(95% CI), 0.1%, 1.1%]; anti-HBc, 4.2% (3.0%, 5.4%); HBsAg, 0.9% (0.4%, 1.6%); anti-HCV, 0.6% (0.1%, 1.1%), and syphilis 1.2% (0.5%, 1.9%). Overall, 6.1% had at least one positive serological marker for any of the tested infections. A majority (87.9%) was sexually active, of whom 12.1% reported a previously diagnosed sexually transmitted infection (STI) and 1.4% a history of commercial sex work. Variables independently associated with any positive serological test included: older age (> or = 25 vs. < 25 years), low monthly income (< or = 4x vs. > 4x minimum wage), previously diagnosed STI, > or = 1 sexual partner, and any illicit drug use. CONCLUSIONS: These are the first population-based estimates of the prevalence of exposure to these infectious diseases and related risks in young women, a population for whom there is a scarcity of data in Brazil. PMID- 18401701 TI - An online needs assessment of a virtual community: what men who use the internet to seek sex with men want in Internet-based HIV prevention. AB - As part of a study to develop effective Internet-based HIV prevention interventions for Men who use the Internet to seek Sex with Men (MISM), we sought information from the target population on; (a) acceptability of sexually explicit media; (b) interest in specific content areas; and (c) identification of credible sources of information. A cross-sectional stratified Internet-based survey design was employed. Between September and November 2005, we recruited 2,716 MISM through Gay.com stratified across race/ethnicity to ensure adequate racial/ethnic diversity. Sixteen Likert-type items assessed acceptability of sexual explicitness, 24 items identified topics for inclusion, and two assessed sources of information. There was near universal acceptability for highly sexually explicit education. Over 75% reported high interest in 10 sexual health topics. HIV positive MISM and MISM engaged in unprotected anal sex with multiple male partners reported significantly less interest in HIV prevention specific content. Differences across age, race/ethnicity and education were identified. Idiosyncratic searches and gay sites were frequently cited sources of information; however blogs, government, and media sites were not. It is acceptable for web-based HIV prevention for MISM to be highly sexually explicit and to provide detailed content relevant to men's sexual health. Since demographic differences in acceptability and content were minor, it is appropriate for interventions to target across demographics. Interventions to re engage men engaging in high risk and HIV + MISM should be considered. Leading health agencies should review whether their web information is retrievable, credible and useful to those most at risk. PMID- 18401702 TI - Poor usage of HUGO standard gene nomenclature in breast cancer studies. PMID- 18401703 TI - Correlates of circulating C-reactive protein and serum amyloid A concentrations in breast cancer survivors. AB - INTRODUCTION: Inflammatory status may be an important prognostic factor for breast cancer. Correlates of markers of inflammation in breast cancer survivors have not been thoroughly evaluated. METHODS: Using data from, the Health, Eating, Activity, and Lifestyle (HEAL) Study (a population-based, multiethnic prospective cohort study of female breast cancer patients) we evaluated the associations between circulating markers of inflammation (C-reactive protein [CRP] and serum amyloid A [SAA], measured approximately 31 months after diagnosis) and several demographic, lifestyle, and clinical characteristics in 741 disease-free breast cancer survivors. Analysis of variance and regression methods were used for statistical analyses of log-transformed values of CRP and SAA. RESULTS: After adjusting for age, BMI, ethnicity, and study site, higher concentrations of CRP were associated with increasing concentration of SAA (P-trend < 0.0001), increasing age (P-trend < 0.0001), increasing BMI (P-trend < 0.0001), increasing waist circumference (P-trend < 0.0001), positive history of heart failure (P = 0.0007), decreasing physical activity (P-trend = 0.005), Hispanic ethnicity (P = 0.05 vs. non-Hispanic white), and current smoking (P = 0.03 vs. never smoking). Vitamin E supplementation (P = 0.0005), tamoxifen use (P = 0.008), and radiation treatment (compared to no chemotherapy or radiation; P = 0.04) were associated with reduced CRP. Associations of CRP with clinical characteristics were not significant in the adjusted models. In a multivariate analysis, CRP showed significant associations with waist circumference, BMI, age, history of heart failure, tamoxifen use, and vitamin E supplementation (R (2) = 0.35). Similar, yet fewer, associations were observed for SAA (R (2) = 0.19). CONCLUSIONS: This study highlights important correlates of inflammatory status in breast cancer patients. Our results are consistent with those from similar studies of healthy women. PMID- 18401705 TI - Enhanced trafficking of tetrameric Kv4.3 channels by KChIP1 clamping. AB - The cytoplamsic auxiliary KChIPs modulate surface expression and gating properties of Kv4 channels. Recent co-crystal structure of Kv4.3 N-terminus and KChIP1 reveals a clamping action of the complex in which a single KChIP1 molecule laterally binds two neighboring Kv4.3 N-termini at different locations, thus forming two contact interfaces involved in the protein-protein interaction. In the second interface, it functions to stabilize the tetrameric assembly, but the role it plays in channel trafficking remains elusive. In this study, we examined the effects of KChIP1 on Kv4 protein trafficking in COS-7 cells expressing EGFP tagged Kv4.3 channels using confocal microscopy. Mutations either in KChIP1 (KChIP1 L39E-Y57A-K61A) or Kv4.3 (Kv4.3 E70A-F73E) that disrupt the protein protein interaction within the second interface can reduce surface expression of Kv4 channel proteins. Kv4.3 C110A, the Zn2+ binding site mutation in T1 domain, that disrupts the tetrameric assembly of the channels can be rescued by WT KChIP1, but not the KChIP1 triple mutant. These results were further confirmed by whole cell current recordings in oocytes. Our findings show that key residues of second interface involved in stabilizing tetrameric assembly can regulate the channel trafficking, indicating an intrinsic link between tetrameric assembly and channel trafficking. The results also suggest that formation of octameric Kv4 and KChIP complex by KChIPs clamping takes place before their trafficking to final destination on the cell surface. PMID- 18401704 TI - Anemia treatment in chronic kidney disease: shifting uncertainty. AB - Epidemiologic observations showing associations between higher levels of some biologic markers such as blood pressure and serum cholesterol with heightened risk of death and non-fatal cardiovascular events have provided important data to develop hypotheses regarding pharmacologic therapies to modify these markers to improve prognosis. Randomized controlled trials have shown that strategies to reduce blood pressure with a variety of antihypertensive agents and LDL cholesterol with statins do, indeed, result in important improvements in clinical outcomes. However, there are several instances where a hypothesis based on strong observational data has been rejected based on surprising counterintuitive evidence generated from randomized controlled clinical trials. Use of inotropic therapies for patients with reduced left ventricular ejection fraction heart failure, administration of class I antiarrhythmic agents to suppress ventricular arrhythmias in high-risk patients, and use of hormone replacement therapy for postmenopausal women have each shown that therapies presumed to be of benefit may actually be producing unfavorable clinical results. Use of erythropoietic stimulating agents (ESA) in chronic kidney disease patients with anemia is similarly based on strong observational data indicating that the degree of anemia is independently associated with higher risk for cardiovascular morbidity and mortality. In non-dialysis patients with mild to moderate anemia, current clinical outcome studies have only addressed arbitrary hemoglobin targets for ESA therapy and have shown that targeting the higher hemoglobin levels was not associated with the benefit and may even result in harm. This review will outline the importance of having a placebo-controlled trial in this patient population to better assess the risk benefit profile of this therapy. PMID- 18401706 TI - A differential proteomic approach reveals an evolutionary conserved regulation of Nme proteins by Fe65 in C. elegans and mouse. AB - The function of the APP-Fe65 complex is still not definitively understood. To address this point we studied the phenotype of Fe65 (feh-1) ablation, which results in severe developmental defects in C. elegans, including embryonic and larval arrests. To shed light on the complex phenotype of embryonic arrest, we undertook a systematic approach, aiming at the definition of the altered proteomic profile of feh-1 null worms. We defined a panel of 27 regulated proteins, 16 of which actually participating to embryonic development processes in the nematode. Protein spots corresponding to the products of the F25H2.5 gene, the nematode orthologue of mammalian Nm23/Nme gene family members, were consistently up-regulated in feh-1 -/- embryos. We observed similar up-regulation of Nme1 and Nme2 genes, both at the transcript and the protein levels, in the brain of Fe65 knock-out mice, thus highlighting the occurrence of evolutionary conserved mechanisms of Nme expression in nematodes and mammals. PMID- 18401707 TI - Role of ciliary neurotrophic factor in microglial phagocytosis. AB - Microglia, CNS-resident macrophages, serve as scavengers to remove cellular debris and facilitate tissue remodeling in the developing and injured CNS. Little is known as what and how microenvironmental factors mediate the phagocytotic ability of microglia. Our previous study has indicated that treatment with glial cell line-derived neurotrophic factor (GDNF) increased the phagocytotic activity of primary rat microglia possibly through the upregulation of alpha5 integrin. In the present study, ciliary neurotrophic factor (CNTF), which has been reported to be produced by glia, was shown to have stimulatory effect on the phagocytosis of primary rat microglia and mouse microglial cell line BV2. Ca2+ imaging analysis and the application of intracellular calcium chelator BAPTA-AM revealed that CNTF induced increase in microglial phagocytosis was mediated by a calcium signaling pathway. Furthermore, treatment with CNTF led to an increase in the expression of alphav integrin, which has been reported to be involved in the phagocytosis of the apoptotic cells. In summary, we have provided evidence that CNTF can increase microglial phagocytosis through a calcium-mediated pathway. Our results also suggest that the upregulation of alphav integrin by CNTF could be involved in the increased phagocytotic activity of microglia. PMID- 18401708 TI - Electro-acupuncture reduces neuronal apoptosis linked to Bax and Bcl-2 expression in the spinal cords of cats subjected to partial dorsal root ganglionectomy. AB - While electro-acupuncture (EA) has been well known to contribute towards neuroplasticity occurring in both the central and the peripheral nervous system after injury, the underlying mechanism remains largely unknown. This study evaluated the effects and the possible mechanism of EA on neuronal apoptosis in the spinal cords of cats subjected to the removal of L(1)-L(5) and L(7)-S(2) dorsal root ganglion, sparing the L(6) dorsal root ganglion. EA treatment decreased the number of TUNEL-positive apoptotic cells in lamina II of the L(3) and L(6) cord segments at 7 and 14 days post operation (dpo). This EA-mediated neuroprotection is associated with a decrease in the number of Bax immunoreactive neurons and an increase in the number of Bcl-2 immunoreactive neurons. Furthermore, Western blot and RT-PCR analysis revealed a significant downregulation of Bax protein and its mRNA, but an upregulation of Bcl-2 in the dorsal horn of L(3) and L(6) cords at both 7 and 14 dpo. The present findings suggest that EA could inhibit neuronal apoptosis in dorsal root deafferentated cat spinal cords, possibly by Bax downregulation and Bcl-2 upregulation. PMID- 18401710 TI - Consumer perspectives on information and other inputs to decision-making: implications for evidence-based practice. AB - This study is an exploration of mental health consumers' perspectives on information, including scientific information, and on other inputs to decision making. Four focus groups were held with severely mentally ill consumers at two sites in the summer of 2005. Consumers varied in age, race and diagnosis. Participant responses were coded by theme and into subthematic categories. Implications for evidence-based decision-making included that: consumers desire and seek information about their illnesses and the mental health system; consumers identify scientific studies as information with special and welcome properties; and consumers also identify other influences on their decision making, most of which fall under the "recovery" rubric. PMID- 18401709 TI - Searching for molecular players differentially involved in neurotransmitter and neuropeptide release. AB - Neurotransmitters and neuropeptides are stored in small clear vesicles (SCVs) and large dense core vesicles (LDCVs), respectively. Many differences in the properties of SCVs and LDCVs suggest that these two classes of secretory organelles may employ different sets of molecules in exocytosis. Relatively little is known, however, about factors that differentially participate in SCVs and LDCVs release. This article briefly overviews some key molecules that are possibly involved in the differential regulation of the trafficking, docking, priming and fusion of SCVs and LDCVs. PMID- 18401711 TI - Longitudinal patterns of health system retention among veterans with schizophrenia or bipolar disorder. AB - Inconsistent service use for schizophrenia and bipolar disorder is associated with poorer outcomes of care. We analyzed VHA National Psychosis Registry data for 164,150 veterans with these disorders to identify characteristics associated with 5-year patterns of survival and with retention in VHA care. Most cohort members (63%) survived the period with no break in VHA healthcare lasting over 12 months. Inconsistent utilization was associated with younger age, no service connected disability, and less physical comorbidity, regardless of diagnosis. The influence of gender and ethnicity on attrition varied by diagnosis and gap duration. Variation in attrition by gender and ethnicity warrants additional attention. PMID- 18401712 TI - The relationship between maternal and child symptom change in community mental health. AB - The mental health of mothers and children are closely linked. This study examined the relationship between child and maternal symptom change during a period in which children participated in community mental health treatment. Symptom change was measured using the Child Behavior Checklist for children and the Beck Depression Inventory for Mothers. Results indicate that mothers whose children improve in community mental health treatment are significantly more likely to report a reduction in maternal depressive symptoms than mothers whose children do not improve. Implications of these findings for mental health service settings are discussed. PMID- 18401713 TI - Factors associated with early attrition from psychosocial rehabilitation programs. AB - This study aimed to identify characteristics associated with early dropout from a vocationally oriented psychosocial rehabilitation (PSR) program for clients with severe mental illness. The sample consisted of 194 individuals who participated in a study comparing a supported employment program to a stepwise vocational program. Study participants who dropped out of the PSR program within 6 months of study entry were compared to those who continued for at least 6 months. Dropouts had poorer competitive employment outcomes than those who continued. Participants with at least a high school diploma, never married, with a schizophrenia-spectrum diagnosis, and those assigned to a stepwise model of vocational rehabilitation were more likely to dropout. The implications of these findings are discussed. PMID- 18401715 TI - Variceal bleeding and long-acting octreotide: a new addition to the armamentarium? PMID- 18401714 TI - Endoscopic evaluation in gastrointestinal graft-versus-host disease: comparisons with histological findings. AB - The gastrointestinal (GI) tract is the major target site of the graft-versus-host disease (GVHD). Whether endoscopic findings can predict the histological diagnosis and degree of severity in GVHD remains controversial. We investigated the degree of correlation between endoscopic and histological findings, and evaluated the impact of endoscopic examination on clinical decision in GVHD management. This study was conducted as a retrospective single-center study. One hundred and one patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) and at the risk of GI GVHD were referred for endoscopic evaluation. Endoscopic images and histology were reviewed to diagnose and grade in a blind fashion, and the histological findings were used as the "gold standard" for diagnosis. Endoscopic findings revealed a significantly positive association with histological abnormalities of GVHD (odds ratio [OR] = 33.6, 95% CI 4.3-261.1), and the sensitivity and specificity were 98 and 44%, respectively. The kappa coefficient for agreement between the endoscopic and histological grading was 0.423 (p-value < 0.001). Ten (18%) patients out of 57 histology negative cases were managed successfully as GVHD on the endoscopic finding. Though the overall reliability of endoscopic diagnosis in GVHD is still insufficient in terms of sensitivity and specificity, high-grade cases such as grades 3 or 4 showed specific endoscopic findings to draw a significant agreement with histological findings. Endoscopic examination can give critical information and impose a pivotal impact on clinical decision when the histology is discordant with clinical presentation. PMID- 18401716 TI - Polymorphisms of the ICAM-1 gene are associated with biliary atresia. AB - Inflammation is an important feature of biliary atresia, and recent studies suggest that its occurs in a genetically susceptible host. The intercellular adhesion molecule-1 (ICAM-1) is of paramount importance for the initiation and propagation of various inflammatory conditions. AIM: To determine whether the Glu241Arg polymorphism in the ICAM-1 gene, which impairs inflammatory responses, is associated with biliary atresia. METHODS: Between February 2002 and November 2004, 19 patients (mean age 1 +/- 0.4 years) diagnosed as biliary atresia were included in the study. Thirty-eight children with chronic liver disease and a group of unrelated healthy controls (n = 123) included in this study. After informed consent, blood was collected and genomic DNA was obtained. Genotyping was performed by amplification-refractory mutation system polymerase chain reaction (ARMSPCR). Associations were assessed by using Fischer's exact test. RESULTS: ICAM G242R A allele frequency was significantly higher in the BA group than in both the CLD and healthy control groups (OR = 4.4, 95 CI% 1.3-15.1, P = 0.03 and OR = 4.8 CI% 1.5-15.6, P = 0.01, respectively). Univariate analysis showed that polymorphism of ICAM G241R polymorphism was significantly related to biliary atresia. There was not significant correlation between PELD score and ICAM-1 genotypes both in BA and CLD groups. CONCLUSION: These findings provide evidence for the possible role of ICAM-1 241R polymorphism in BA pathogenesis. PMID- 18401717 TI - Differences in the influence of family background and social activities on smoking of minority and white high school seniors, 1976-2004. AB - This population-based study of high school seniors examines differences in the influence of several important determinants of smoking among white, African American, Hispanic, and other teens. With national survey data for each year from 1976 to 2004, logistic regression is used to test for differences across race and ethnic groups (averaged over all years) in the effects on daily smoking of background factors such as living arrangements and parents' education and social activities such as academic performance and religiosity. The results show similarity in effects across racial categories, but some determinants have weaker or reversed effects for African-American and Hispanic youth than for white youth. For example, high parents' education increases smoking among Hispanic youth, has little influence among African Americans, and decreases smoking among whites. These results suggest that smoking patterns of minority teens differ in some ways from those of white teens. PMID- 18401718 TI - Human tissue factor pathway inhibitor-2 induces caspase-mediated apoptosis in a human fibrosarcoma cell line. AB - Human TFPI-2 is an extracellular matrix-associated Kunitz-type serine proteinase inhibitor. We previously demonstrated that a human fibrosarcoma cell line, HT 1080, does not express TFPI-2, but genetic restoration of TFPI-2 expression in these cells markedly inhibited their growth and metastasis in vivo. In the present study, either full-length recombinant TFPI-2, or its mutated first Kunitz type domain (R24K KD1), were offered to HT-1080 cells, and the degree of apoptosis assessed by nuclear fragmentation, ethidium bromide and acridine orange staining, fluorescence activated cell sorting, immunoblotting and gene expression profiling. R24K KD1 induced apoptosis in 69% of HT-1080 cells in a 48 h period compared to 39% for TFPI-2, while a KD1 preparation lacking a reactive site arginine/lysine residue (R24Q KD1) produced only an 18% apoptosis rate, suggesting that the observed apoptosis was related to proteinase inhibition. Immunoblotting experiments indicated increased caspase 3 and 9 activation, up regulation of pro-apoptotic Bax and suppression of anti-apoptotic Bcl-2 protein. Finally, microarray analyses of R24K KD1-treated cells indicated elevated expression of several pro-apoptotic genes and under-expression of anti-apoptotic genes. Collectively, our results demonstrate that treatment of HT-1080 cells exogenously with either TFPI-2 or R24K KD1 activates caspase-mediated, pro apoptotic signaling pathways resulting in apoptosis. PMID- 18401719 TI - Adenovirus-mediated PDCD5 gene transfer sensitizes K562 cells to apoptosis induced by idarubicin in vitro and in vivo. AB - PDCD5 (programmed cell death 5) accelerates apoptosis of certain tumor cells and is expressed at low levels in marrow-nucleated cells of AML and CML patients. In the present study, we evaluated the effects of PDCD5 overexpression on drug sensitivity of leukemia cells. K562 cells were treated with idarubicin (IDR) alone or in combination with adenoviral vectors expressing PDCD5 (Ad-PDCD5). As shown by annexin-V-FITC/PI dual labeling, apoptosis rates were markedly increased after combined treatment with Ad-PDCD5 compared to IDR treatment alone. We observed that PDCD5 overexpression significantly improves the antitumor effects of low dose IDR treatment in vivo. Tumor sizes were significantly decreased in combined Ad-PDCD5 and low dose IDR treatment groups compared with single IDR treatment groups. Similar results were obtained with combined systemic treatment of Ad-PDCD5 and low dose IDR, and combined treatment with Ad-PDCD5 local injection and low dose IDR i.p. injection. These results indicate that Ad-PDCD5 may be a promising agent for enhancing chemosensitivity. PMID- 18401720 TI - Lymphocytic hypophysitis with associated thyroiditis in a man with aseptic meningitis. AB - OBJECTIVE: Lymphocytic hypophysitis (LH) is a rare chronic inflammatory disorder characterized by lymphocytic infiltration of the pituitary gland commonly affecting women during pregnancy or post-partum period. The pathogenesis remains uncertain, however an autoimmune process is frequently implicated. There is limited data on the occurrence of LH outside the setting of autoimmunity. CASE: We describe a 37-year-old man presenting with diarrhoea, nausea, weight loss, low grade fever, headache and cerebrospinal fluid analysis consistent with aseptic meningitis. Magnetic resonance imaging (MRI) demonstrated a homogenously enlarged pituitary gland with biochemical testing revealing partial hypopituitarism with adenocorticotrophic hormone and gonadotrophin deficiency. Notably, his free thyroid hormone levels were elevated with a suppressed thyroid-stimulating hormone and a suppressed thyroid technetium scan consistent with thyroiditis. Tissue antibodies including thyroid antibodies were negative. Following introduction of hydrocortisone, he developed transient diabetes insipidus which spontaneously resolved after 4 months. Thyrotoxicosis resolved after 5 weeks and thyroxine was commenced as he developed secondary hypothyroidism. Repeat MRI 3 months later showed a reduction in the size of the pituitary gland which by 6 months had returned to normal size. He remains well on hydrocortisone, thyroxine and testosterone replacement. CONCLUSIONS: Based on clinical and radiological grounds, the diagnosis was consistent with lymphocytic hypophysitis associated with subacute thyroiditis. This is only the second report of this combination in the absence of autoimmunity and the first report of LH and thyroiditis with associated aseptic meningitis in the absence of tissue autoantibodies. We propose a possible viral illness as the unifying aetiological cause. PMID- 18401721 TI - Syncytial coupling of neurons in tissue culture and early ontogenesis. AB - Computerized time-lapse video recording was used to detect the process of formation of syncytial couplings between the processes of different neurons in dissociated neuron cultures. These studies showed that once the processes of one neuron had formed connections with another neuron, death of the cell body (its trophic center) was not followed by Wallerian degeneration. Translocation of cytoplasmic varicosities along the branches of one neuron to another was observed over periods of several hours. Electron microscopic studies of the nerve processes of cells in the intramural intestinal plexus in the early postnatal period demonstrated all the transitional states from fusion and perforation of the membranes of contacting dendrites to complete fusion of the neuroplasm of processes with formation of residual membranous structures at the location of the former intercellular contact. PMID- 18401723 TI - Quantitative analysis of the distribution of the motor cortex representations of the fore-and hindlimbs in the red nucleus of the cat. AB - A quantitative analysis of the distribution of corticorubral fibers was performed after precise electrolytic lesioning of the lateral and medial margins of the posterior sigmoid gyrus--the motor representations of the fore-and hindlimbs respectively--in cats. The cortical representations of the forelimbs were found to project to the whole of the rostrocaudal extent of the red nucleus (RN). The number of efferent fibers terminating at the rostral margin of the RN was almost twice that terminating in the caudal third of the RN. Efferent fibers of the cortical representation of the hindlimbs did not project to the rostral two thirds of the RN but ended in its caudal third; the number of projecting corticorubral fibers was the same as the number running from the cortical representation of the forepaws to the caudal third of the RN. The significantly (almost double) greater number of fibers running from the cortical representation of the forelimbs in comparison with the number directed from the representation of the hindlimbs found in the present study is probably evidence of the greater functional importance of corticorubral connections in movement reactions performed by the forelimbs. PMID- 18401722 TI - Morphological characteristics of apoptosis and its significance in neurogenesis. AB - Results from our own studies and published data are used to provide a critical analysis of current views of the role of apoptosis in regulating the quantitative and qualitative constancy of cells in the developing brain. Detailed descriptions of the morphological features and mechanisms of the different phases of apoptotic neuron death are given. Apoptosis affects the ordering of connections in neural networks and is involved in the pathogenesis of neurodegenerative diseases- epilepsy, schizophrenia, and Alzheimer's disease. The question of the interaction of the NO-ergic mechanism and the cell in executing apoptosis is discussed. The data allow NO to be regarded as a cytotoxic factor which induces apoptosis. The ultrastructural features of post-mitotic immature neurons suggests the operation of the apoptotic and necrosis-like (NO-dependent) pathways of natural cell death during the process of differentiation. PMID- 18401724 TI - Morphofunctional state of neurons in the temporal cortex and hippocampus in relation to the level of spatial memory in rats after ablation of the olfactory bulbs. AB - Ablation of the olfactory bulbs (bulbectomy) in mice and guinea pigs evokes a neurodegenerative process which, in terms of its morphological, biochemical, and behavioral features, is similar to Alzheimer's disease. We report here studies of the long-term sequelae of bulbectomy in rats. One year after surgery, testing of spatial memory in bulbectomized rats (BER) allowed the animals to be divided into two groups-those with good memory (BER-gm) and those with poor memory (BER-pm). Quantitative analysis of the morphofunctional state of neurons showed that BER pm, as compared with the BER-gm group, had more marked pathological lesions in neurons of the temporal cortex and hippocampus, with significant increases in the numbers of cells showing pyknosis, karyolysis, cytolysis, and vacuolization. Both groups showed decreases in the distribution density of cells in the cortex. In terms of the level of brain beta-amyloid, the study groups fell in the order: BER pm>BER-gm>control sham-operated rats. These results provide evidence of the long term nature of changes in the morphofunctional state of neurons in the brains of BER, correlating with their levels of spatial memory. PMID- 18401725 TI - Neuron density in the hippocampus in rat strains with contrasting nervous system excitability after prolonged emotional-pain stress. AB - A model system consisting of two rat strains bred for nervous system excitability in response to electric shocks was used to study changes in the number density of neurons in hippocampal field CA3 at 24 h, two weeks, and two and six months after prolonged emotional-pain stress (PEPS). Neuron density in hippocampal field CA3 decreased after completion of PEPS. These changes arose at different time points in the different rat strains (one day for low-excitability rats, two months for high-excitability rats) and persisted to six months. Thus, this is the first demonstration that persistent differential effects of stress on the number density of neurons in hippocampal field CA3, which plays an important role in learning and memory processes, depend on genetically determined constitutive characteristics of the nervous system. PMID- 18401726 TI - Kindling-induced hippocampal cell death in rats. AB - Quantitative analysis of the main cells and interneurons in different layers and fields of Ammon's horn, the hilus, and dentate gyrus was performed two weeks and one month after kindling, i.e., specific electrical stimulation of the ventral hippocampus. At both experimental time points, cells numbers were significantly decreased at all levels of the areas studied except the marginal layer of field CA4. Thus, in this model of experimental epilepsy, the process of epileptogenesis involved both interneurons and main cells in the hippocampus. The possible reorganization of intrahippocampal circuits and their involvement in the increased excitability characteristic of epilepsy are discussed. PMID- 18401727 TI - Neuron-glial contacts formed in the cerebellum during electrical stimulation in the presence of an NO-generating compound. AB - The molecular layer of the cerebellum of the frog Rana temporaria was studied by light and electron microscopy after electrical stimulation in the presence of an NO-generating compound. In these conditions, there was severe swelling of granule cell axon terminals (boutons) and astrocyte processes (AP), with loss of cytoplasmic elements. However, along with damaged structures, there were also undamaged structures: boutons with synaptic vesicles and AP with glycogen granules. It is suggested that these persisting viable AP may form 1) glial "wrappings" around damaged synapses or boutons and 2) neuron-glial contacts, which form when synaptic vesicles cross damaged bouton membranes to AP containing glycogen granules. It is also suggested that the presence of glycogen in AP in conditions of oxygen and glucose deficiency may provide a source of high-energy substrates such as glucose and ATP, thus providing conditions for neuron survival in pathological states (ischemia/hypoxia). PMID- 18401728 TI - Comparative analysis of the locations of the NR1 and NR2 NMDA receptor subunits in honeybee (Apis mellifera) and fruit fly (Drosophila melanogaster, Canton-S wild-type) cerebral ganglia. AB - The locations of the NR1 and NR2 subunits of the GABA receptor were studied in brain structures in insects--honeybees and fruit flies--using an immunohistochemical method. The specificities of the antibodies to the NR1 and NR2 subunits were confirmed by the antisense knockdown method for the NR1 subunit and western blotting. The data obtained here lead to the conclusion that the distributions of the NR1 and NR2 subunits of the NMDA receptor complex in the cerebral ganglia of the honeybee and fruit fly are similar; areas with the highest concentrations of NR1 and NR2 subunits were identified, and these were found to be different in the different insects. This is associated with the behavioral characteristics of these two insect species. PMID- 18401729 TI - Development of brain vessels in human embryos and fetuses in conditions of prenatal exposure to alcohol. AB - Light and electron microscopy were used to study the characteristics of the formation of brain vascular structures at the early stages of development in conditions of maternal alcoholization during pregnancy. Computer morphometric methods using the Scion Image system for image analysis showed that fetuses at 11 12 weeks of development in conditions of prenatal alcohol exposure showed a decrease in the mean absolute cross-sectional area of vessels in the intermediate layer of the brain, with an increase in their relative area and an increase per unit area of sections, as compared with the control group. Vessels started to differentiate into arteries and veins from 10 weeks of development. PMID- 18401730 TI - Effects of beta-amyloid (25-35) on learning in the common snail. AB - The effects of neurotoxic beta-amyloid fragment (25-35) on the formation of behavioral sensitization and a conditioned defensive reflex to food were studied. Administration of beta-amyloid (25-35) to common snails before the start of training led to a significant reduction in sensitization of the defensive reaction, weakening of the formation of the conditioned defensive reflex to food, and impairment of memory. These impairments to behavioral plasticity may be mediated by changes in synaptic plasticity previously observed in the presence of beta-amyloid. PMID- 18401731 TI - Involvement of brain dopaminergic systems in the development of an MPTP-induced depressive state in rats. AB - A depressive state was induced in Wistar rats by repeated i.p. injections of the proneurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which induces the death of dopaminergic neurons in the brain. Signs of the development of the experimental depressive state were a reduction in body weight and an increase in the proportion of REM sleep during daytime sleep. The rearrangement of the spectral characteristics of electrical activity during the development of experimental depressive syndrome in rats was shown to occur in target structures of the nigrostriatal, mesocortical, and mesolimbic dopaminergic systems of the brain, as well as in the amygdala and hippocampus. The most marked changes were seen in the terminal field of the nigrostriatal dopaminergic system and hippocampus. Spectral rearrangements of electrical activity in the theta-1 and theta-2 ranges in the hippocampus and dopaminergic structures suggest the involvement of the hippocampus in mediating changes in the emotional status of the experimental animals during the development of the MPTP-induced depressive state. PMID- 18401732 TI - Age-related features of the interaction of learning success and characteristics of auditory operative memory. AB - The characteristics of the interaction between learning success and measures of auditory operative memory were studied by psychoacoustic testing of 42 medical workers aged 20-65 years trained to work in areas new to them (information science). Three age groups were identified: 20-35 years, 36-50 years, and 51-65 years. The acoustic test consisted of a single presentation via headphones of 12 sequential target words from information science with subsequent presentation of 12 target and 12 masking words in random order. The subjects' task was to recognize the target words. Stepwise linear regression analysis identified a relationship between the efficiency with which the new material was learned and measures of auditory operative memory, whose role in learning success increased with age. Since subjects older than 35 years showed a reduction in remembering efficiency, it was suggested that age-related changes in the characteristics of auditory operative memory are one of the major reasons for degradation of the ability to learn new material. PMID- 18401733 TI - The role of dopamine-dependent negative feedback in the hippocampus-basal ganglia thalamus-hippocampus loop in the extinction of responses. AB - A mechanism for the extinction of the responses of hippocampal and dopaminergic neurons to repeated sensory stimuli is proposed, based on dopamine-dependent negative feedback in the hippocampus-basal ganglia-thalamus-hippocampus loop. Activation of hippocampal neurons evoked by a new stimulus facilitates the appearance of responses in dopaminergic neurons as a result of disinhibition via striopallidal cells of the nucleus accumbens and ventral pallidum. However, increases in dopamine levels and activation of D2 receptors on striopallidal cells, facilitating depression of hippocampal inputs, prevent disinhibition of dopaminergic neurons, such that their responses start to decline. Subsequent reductions in actions on D1 receptors lead to decreases in the efficiency of excitation both of neurons in hippocampal field CA1 and strionigral cells in the nucleus accumbens. The direct pathway via the basal ganglia mediates disinhibition of the thalamic nucleus reuniens, exciting neurons in field CA1, which leads to extinction of the responses of hippocampal neurons, decreases in disinhibition of dopaminergic cells, and further extinction of their responses. PMID- 18401734 TI - Modification of the effects of glutamate by nitric oxide (NO) in a pattern generating network. AB - Previous studies have shown that nitric oxide (NO) transforms the responses of various neurons to glutamate, though it remained unclear whether this mechanism is involved in the formation of behavior. We therefore studied the buccal generator of the feeding rhythm of the mollusk Lymnaea stagnalis (pond snail). In this organism, glutamate and NO are synthesized by defined neurons; glutamate is the neurotransmitter for the second phase of the standard triphasic feeding rhythm. Motoneuron B4 was used for monitoring. Studies using isolated CNS preparations showed that in some cases glutamate evoked hyperpolarization of B4 and terminated rhythmic network activity (n=17; group 1), while in other cases glutamate evoked depolarization of B4 and activation of a non-standard biphasic rhythm (n=12; group 2). In group 1, the NO donor nitroprusside lifted the inhibitory effect of glutamate (n=13), with transformation into an excitatory effect in nine cases. In group 2, the NO acceptor PTIO transformed the excitatory effect of glutamate into an inhibitory effect (n=7). These results provide evidence that: 1) the responses of the central generator of the buccal motor rhythm to glutamate depend on the NO level, and 2) this regulatory mechanism can modify feeding behavior. PMID- 18401735 TI - Expression of the c-Fos gene in the rat hypothalamus in electrical pain stimulation and UHF stimulation of the skin. AB - Studies of the pattern of activation of hypothalamic cells in conditions of electrical pain stimulation demonstrated an increase in the proportion of c-Fos positive cells (to 72.02-98.95%), predominantly affecting cells of size 10-50 microm2. UHF irradiation of the skin activated hypothalamic cells of size 10-70 microm2 in the paraventricular and dorsomedial nuclei and in the periforniceal zone of the lateral hypothalamic field. Combined action of UHF irradiation of the skin and electrical pain stimulation led to decreases in the proportions of cells activated as a result of pain stimulation in the anterior hypothalamic field, the ventromedial nucleus, and the basal part of the lateral hypothalamic field. Changes in the distribution of activated cells by size (mainly a reduction in the proportion of cells of area 10-30 microm2 and an increase in the proportion of larger cells) were detected in the ventromedial nucleus and the basal part of the lateral field. UHF irradiation of the skin was found to have a modifying effect, consisting of a reduction in the degree of activation of hypothalamic cells evoked by electrical pain stimulation. PMID- 18401736 TI - Involvement of ionotropic glutamate receptors in the appearance of arecoline tremor in mice. AB - Administration of the muscarinic cholinoreceptor agonist arecoline (6 mg/kg, s.c.) to mice induced long-lasting tremor. The ability of non-competitive antagonists of ionotropic glutamate receptors to suppress the onset of tremor was studied. These antagonists, i.e., adamantane and phenylcyclohexyl derivatives, selectively blocked NMDA-type receptor channels (monocations) or both NMDA-and AMPA-type channels (dications). Both types of blocker weakened arecoline tremor, though the dose-response relationships were different for mono-and dications. The effects of dications appeared only at low blocker doses (0.0001-0.01 micromol/kg) but gradually disappeared on dose elevation. These data lead to the conclusion that the mechanism of pathogenesis of arecoline tremor predominantly involves NMDA-type receptors. Moderate blockade of AMPA-type receptors could potentiate the preventive effect of mixed-action antagonists (anti-NMDA+anti-AMPA), though predominance of blocking action against AMPA-type receptors prevented this effect. PMID- 18401737 TI - Prediction of outbreaks of post-traumatic stress disorder after large-scale terrorist acts. AB - An original computer technology--a mathematical model and computer programs running under Windows--for the operative assessment of outbreaks of post traumatic stress disorders (PTSD) and prediction of their sequelae among victims of terrorist attacks is described. The structure of the mathematical model is described, along with the relationship between the mathematical model and the calculating algorithm for prediction of the development of outbreaks of PTSD. The computer program allowed predictions of the numbers of mental disorders with subsequent behavioral impairment, addictive disorders, personality changes, and signs of maladaptation in victims of terrorist acts. A model of an epidemic of mass panic in a population sensitive to negative information on the consequences of terrorist acts is described. PMID- 18401738 TI - Morphochemical characteristics of neuron responses in functionally different brain structures to amphetamine treatment of different durations. PMID- 18401739 TI - Baby blues syndrome as an adaptation disorder in the early stages of formation of the mother-child system. PMID- 18401740 TI - Heterologous expression of a hyperthermophilic alpha-amylase in xanthan gum producing Xanthomonas campestris cells. AB - A hyperthermophilic alpha-amylase encoding gene from Pyrococcus woesei was transferred and expressed in Xanthomonas campestris ATCC 13951. The heterologous alpha-amylase activity was detected in the intracellular fraction of X. campestris and presented similar thermostability and catalytic properties with the native P. woesei enzyme. The recombinant alpha-amylase was found to be stable at 90 degrees C for 4 h and within the same period it retained more than 50% of its initial activity at 110 degrees C. Furthermore, X. campestris transformants produced similar levels of recombinant alpha-amylase activity regardless of the carbon source present in the growth medium, whereas the native X. campestris alpha-amylase production was highly dependent on starch availability and it was suppressed in the presence of glucose or other reducing sugars. On the other hand, xanthan gum yield, which appeared to be similar for both wild type and recombinant X. campestris strains, was enhanced at higher starch or glucose concentrations. Evidence presented in this study supports that X. campestris is a promising cell factory for the co-production of recombinant hyperthermophilic alpha-amylase and xanthan gum. PMID- 18401741 TI - PCR-based gene synthesis, molecular cloning, high level expression, purification, and characterization of novel antimicrobial peptide, brevinin-2R, in Escherichia coli. AB - Brevinin-2R, a member of a new family of antimicrobial peptides isolated from the skin of Rana ridibunda, displays antimicrobial activity against bacteria and fungi. In this study, we have used an assembly PCR method for the fast and extremely accurate synthesis of the brevinin-2R gene. A total of six primers were assembled in a single step PCR, and the assembly was then amplified by PCR to produce the final gene. The synthetic gene was cloned into the pET32a (+) vector to allow the expression of brevinin-2R as a Trx fusion protein in Escherichia coli. The results indicated that the expression level of the fusion protein could reach up to 25% of the total cell proteins. The expression products could be easily purified by Ni-NTA chromatography and released from the fusion protein by factor Xa protease. The peptide displayed antimicrobial activity similar to that of the purified brevinin that was reported earlier. This method allows the fast synthesis of a gene that optimized the overexpression in the E. coli system and production of sufficiently large amounts of peptide for functional and structural characterizations. PMID- 18401742 TI - Studies on the chemical modification of the essential groups of N-Acetyl-beta-D glucosaminidase from viscera of green crab (Scylla Serrata). AB - The chemical modification of N-acetyl-beta-D: -glucosaminidase (EC3.2.1.30) from viscera of green crab (Scylla serrata) has been first studied. The modification of indole groups of tryptophan of the enzyme by N-bromosuccinimide can lead to complete inactivation, accompanying the absorption decreasing at 275 nm and the fluorescence intensity quenching at 338 nm, indicating that tryptophan is essential residue to the enzyme. The modification of histidine residue, the carboxyl groups, and lysine residue inactivates the enzyme completely or incompletely. The results show that imidazole groups of histidine residue or sulfhydryl residues, the carboxyl groups of acidic amino acid, amino groups of lysine residue, and indole groups of tryptophan were essential for the catalytic activity of enzyme, while the results demonstrate that the disulfide bonds and the carbamidine groups of arginine residues are not essential to the enzyme's function. PMID- 18401743 TI - Application relevant studies of fungal beta-galactosidases with potential application in the alleviation of lactose intolerance. AB - Functional screening studies revealed that Aspergillus carbonarius ATCC6276 produced extracellular beta-galactosidase activity potentially suited for use as a lactase digestive supplement in the treatment of lactose intolerance. The crude preparation contained two beta-galactosidase activities, beta-gal 1 and beta-gal 2, which were separated by ion-exchange chromatography. Both enzymes were purified to homogeneity by a combination of gel filtration, ion-exchange, chromatofocusing and hydrophobic interaction chromatographies. beta-gal 1 and beta-gal 2 displayed differences in molecular mass (110 kDa versus 120 kDa as judged by SDS PAGE) and in a range of additional physicochemical properties. Km values of 83 and 309 mM, respectively, were recorded using lactose as substrate while temperature optima of 55 degrees C versus 65 degrees C were obtained. Unlike current commercialized supplemental lactases, both of the purified enzymes displayed significant stability when exposed to simulated gastric conditions, with beta-gal 1 in particular retaining 70% residual activity after exposure to pH 2.0 in the presence of pepsin for 2 h. Overall the results indicate that the beta-galactosidases of Aspergillus carbonarius ATCC6276, either individually or in combination, may be suitable for use as a digestive supplement for the alleviation of lactose intolerance. PMID- 18401744 TI - Prokaryotic expression, purification and characterization of Aspergillus sulphureus beta-mannanase and site-directed mutagenesis of the catalytic residues. AB - Wild type (WT) DNA sequence, which encoded a mature beta-mannanase of Aspergillus sulphureus, composed of 1,152 nucleotides (nt), was amplified from pUCm-T-mann by polymerase chain reaction (PCR). Based on this DNA fragment, mutants designated as E(206)G and E(314)G were constructed by overextension PCR (OE-PCR). Glutamic acids of the 206th and 314th sites in the amino acid sequence of beta-mannanase were separately replaced by glycine in these two mutants. The WT and mutant genes were ligated into prokaryotic vector pET-28a (+) and transformed into the Escherichia coli BL21 strain, respectively. The recombinant enzyme proteins were expressed by IPTG induction and detected by Western blot. The recombinant proteins purified with Ni-NTA column were dialyzed to correctly refold. The WT recombinant beta-mannanase showed optimal activity at 50 degrees C and pH 2.4. The kinetic parameters of K (m) and V (max) for purified beta-mannanase were 1.38 mg/ml and 72.99 U/mg, respectively. However, the mutant proteins did not show any activity. It was demonstrated that E(206) and E(314) were the catalytic residues of beta-mannanase. PMID- 18401745 TI - Increasing the value of hominy feed as a coproduct by fermentation. AB - Hominy feed is a low value ($83.7/metric ton) coproduct of the corn dry milling process that accounts for nearly 35% of the starting corn quantity. The average composition of hominy feed on a dry basis is 56.9% starch, 25.2% neutral detergent fiber, 11.1% protein, and 5.3% fat. Starch in hominy feed can be fermented to ethanol thus increasing its levels of protein and fat. The increase in protein and fat percentages may increase the market competitiveness and price of hominy feed. Hydrolysis and fermentation were performed on nine hominy feed samples collected from three corn dry milling plants in the USA. The original hominy feed samples and postfermentation solids were analyzed for starch, protein, fat, and fiber content. Compared to the original hominy feed, the percentage increase in protein, fat and fiber in postfermentation solids of nine samples ranged from 10.4 to 21.3, 6.78 to 10.6, and 12.6 to 28.7% (dry basis), respectively. Ethanol yields varied from 271.7 to 380.2 l/metric ton for the nine hominy feed samples. These results indicate that the value of hominy feed as an animal feedstock can potentially be increased with fermentation and can produce more profit per metric ton than currently being derived by its sale as a low protein feed ingredient. PMID- 18401746 TI - Isolation of novel alkaliphilic bacillus strains for cyclodextrin glucanotransferase production. AB - New alkaliphilic Bacillus producers of cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) were isolated from 17 Bulgarian alkaline and normal habitats (springs and soils) by three steps of a selection. None of the isolates obtained, producing CGTase, appeared to be thermophilic in character. One hundred and thirty-seven strains were estimated for CGTase activity by batch cultivation in a liquid alkaline medium. Twenty-seven of them had a detectable CGTase activity in their culture supernatants under the enzyme assay conditions, despite of the significant growth of all isolates. The phenotypic properties of three selected strains (20RF, 8SB and 24WE) were determined. They were aerobic endospore-forming Bacillus strains: two of them were obligated alkaliphiles (20RF and 8SB) and one, alkalitolerant (24WE). Both obligated alkaliphiles were further characterised by 16S rRNA analysis. According to the full 16S rRNA gene sequences obtained and deposited to the NCBI GenBank database, both isolated obligated alkaliphiles 20RF and 8SB were clustered into the group of alkaliphilic Bacillus species. The exhibited CGTase production by them (230-250 U ml(-1) for 20RF and 130-160 U ml( 1) for 8SB) defined these new isolates as promising producers of the enzyme, especially Bacillus sp. 8SB synthesising thermostable alkaline beta-CGTase. Both new enzymes from 20RF and 8SB Bacillus strains formed only two types of cyclodextrins, beta and gamma, which could be of interest for their easy separation and industrial production. PMID- 18401747 TI - Purification and biochemical characterization of extracellular beta-glucosidases from the hypercellulolytic Pol6 mutant of Penicillium occitanis. AB - The Pol6 mutant of Penicillium occitanis fungus is of great biotechnological interest since it possesses a high capacity of cellulases and beta-glucosidase production with high cellulose degradation efficiency (Jain et al., Enzyme Microb Technol, 12:691-696, 1990; Hadj-Taieb et al., Appl Microbiol Biotechnol, 37:197 201, 1992; Ellouz Chaabouni et al., Enzyme Microb Technol, 16:538-542, 1994; Ellouz Chaabouni et al., Appl Microbiol Biotechnol, 43:267-269, 1995). In this work, two forms of beta-glucosidase (beta-glu 1 and beta-glu 2) were purified from the culture supernatant of the Pol6 strain by gel filtration, ion exchange chromatography, and preparative anionic native electrophoresis. These enzymes were eluted as two distinct species from the diethylamino ethanol Sepharose CL6B and anionic native electrophoresis. However, both behaved identically on sodium dodecyl sulfate polyacrylamide gel electrophoresis (MW, 98 kDa), shared the same amino acid composition, carbohydrate content (8%), and kinetic properties. Moreover, they strongly cross-reacted immunologically. They were active on cellobiose and pNPG with Km values of 1.43 and 0.37 mM, respectively. beta-glu 1 and beta-glu 2 were competitively inhibited by 1 mM of glucose and 0.03 mM of delta-gluconolactone. They were also significantly inhibited by Hg(2+) and Cu(2) at 2 mM. The addition of purified enzymes to the poor beta-glucosidase crude extract of Trichoderma reesei increased its hydrolytic efficiency on H(3)P0(4) swollen cellulose but had no effect with P. occitanis crude extract. Besides their hydrolytic activities, beta-glu 1 and beta-glu 2 were endowed with trans glycosidase activity at high concentration of glucose. PMID- 18401748 TI - Phytase production by a marine yeast Kodamea ohmeri BG3. AB - The marine yeast strain Kodamea ohmeri BG3 isolated from the gut of a marine fish (Hexagrammes otakii) was found to secrete a large amount of phytase into the medium. The crude phytase produced by this marine yeast showed the highest activity at pH 5.0 and 65 degrees C. The optimal medium for phytase production contained oat 10.0 g/l, ammonium sulfate 15.0 g/l, glucose 30 g/l, and NaCl 20.0 g/l, while the optimal cultivation conditions for phytase production were pH 5.0, a temperature of 28 degrees C, and a shaking speed of 170 rpm. Under the optimal conditions, over 557.9 mU/ml of phytase activity was produced within 72 h of fermentation at the shake flask level. This is a very high level of phytase activity produced by yeasts. We think that the medium and process for phytase production by the marine yeast strain were very simple, and such marine yeast from the gut of natural marine fish may have a potential application in the maricultural industry and marine environmental protection. The results demonstrate that phytate was actively degraded by the crude phytase within a short period. PMID- 18401749 TI - Separate and concentrate lactic acid using combination of nanofiltration and reverse osmosis membranes. AB - The processes of lactic acid production include two key stages, which are (a) fermentation and (b) product recovery. In this study, free cell of Bifidobacterium longum was used to produce lactic acid from cheese whey. The produced lactic acid was then separated and purified from the fermentation broth using combination of nanofiltration and reverse osmosis membranes. Nanofiltration membrane with a molecular weight cutoff of 100-400 Da was used to separate lactic acid from lactose and cells in the cheese whey fermentation broth in the first step. The obtained permeate from the above nanofiltration is mainly composed of lactic acid and water, which was then concentrated with a reverse osmosis membrane in the second step. Among the tested nanofiltration membranes, HL membrane from GE Osmonics has the highest lactose retention (97 +/- 1%). In the reverse osmosis process, the ADF membrane could retain 100% of lactic acid to obtain permeate with water only. The effect of membrane and pressure on permeate flux and retention of lactose/lactic acid was also reported in this paper. PMID- 18401750 TI - Parameter estimation for simultaneous saccharification and fermentation of food waste into ethanol using Matlab Simulink. AB - The increase in waste disposal and energy costs has provided an incentive to convert carbohydrate-rich food waste streams into fuel. For example, dining halls and restaurants discard foods that require tipping fees for removal. An effective use of food waste may be the enzymatic hydrolysis of the waste to simple sugars and fermentation of the sugars to ethanol. As these wastes have complex compositions which may change day-to-day, experiments were carried out to test fermentability of two different types of food waste at 27 degrees C using Saccharomyces cerevisiae yeast (ATCC4124) and Genencor's STARGEN enzyme in batch simultaneous saccharification and fermentation (SSF) experiments. A mathematical model of SSF based on experimentally matched rate equations for enzyme hydrolysis and yeast fermentation was developed in Matlab Simulink. Using Simulink parameter estimation 1.1.3, parameters for hydrolysis and fermentation were estimated through modified Michaelis-Menten and Monod-type equations with the aim of predicting changes in the levels of ethanol and glycerol from different initial concentrations of glucose, fructose, maltose, and starch. The model predictions and experimental observations agree reasonably well for the two food waste streams and a third validation dataset. The approach of using Simulink as a dynamic visual model for SSF represents a simple method which can be applied to a variety of biological pathways and may be very useful for systems approaches in metabolic engineering in the future. PMID- 18401751 TI - Oxygen-controlled biosurfactant production in a bench scale bioreactor. AB - Rhamnolipids have been pointed out as promising biosurfactants. The most studied microorganisms for the aerobic production of these molecules are the bacteria of the genus Pseudomonas. The aim of this work was to produce a rhamnolipid-type biosurfactant in a bench-scale bioreactor by one strain of Pseudomonas aeruginosa isolated from oil environments. To study the microorganism growth and production dependency on oxygen, a nondispersive oxygenation device was developed, and a programmable logic controller (PLC) was used to set the dissolved oxygen (DO) concentration. Using the data stored in a computer and the predetermined characteristics of the oxygenation device, it was possible to evaluate the oxygen uptake rate (OUR) and the specific OUR (SOUR) of this microorganism. These rates, obtained for some different DO concentrations, were then compared to the bacterial growth, to the carbon source consumption, and to the rhamnolipid and other virulence factors production. The SOUR presented an initial value of about 60.0 mgO(2)/g(DW) h. Then, when the exponential growth phase begins, there is a rise in this rate. After that, the SOUR reduces to about 20.0 mgO(2)/g(DW) h. The carbon source consumption is linear during the whole process. PMID- 18401752 TI - Continuous production of ethanol from starch using glucoamylase and yeast co immobilized in pectin gel. AB - This work presents a continuous simultaneous saccharification and fermentation (SSF) process to produce ethanol from starch using glucoamylase and Saccharomyces cerevisiae co-immobilized in pectin gel. The enzyme was immobilized on macroporous silica, after silanization and activation of the support with glutaraldehyde. The silica-enzyme derivative was co-immobilized with yeast in pectin gel. This biocatalyst was used to produce ethanol from liquefied manioc root flour syrup, in three fixed bed reactors. The initial reactor yeast load was 0.05 g wet yeast/ml of reactor (0.1 g wet yeast/g gel), used in all SSF experiments. The enzyme concentration in the reactor was defined by running SSF batch assays, using different amount of silica-enzyme derivative, co-immobilized with yeast in pectin gel. The chosen reactor enzyme concentration, 3.77 U/ml, allowed fermentation to be the rate-limiting step in the batch experiment. In this condition, using initial substrate concentration of 166.0 g/l of total reducing sugars (TRS), 1 ml gel/1 ml of medium, ethanol productivity of 8.3 g/l/h was achieved, for total conversion of starch to ethanol and 91% of the theoretical yield. In the continuous runs, feeding 163.0 g/l of TRS and using the same enzyme and yeast concentrations used in the batch run, ethanol productivity was 5.9 g ethanol/l/h, with 97% of substrate conversion and 81% of the ethanol theoretical yield. Diffusion effects in the extra-biocatalyst film seemed to be reduced when operating at superficial velocities above 3.7 x 10(-4) cm/s. PMID- 18401753 TI - Lipase production in solid-state fermentation monitoring biomass growth of aspergillus niger using digital image processing. AB - The aim of this study was to monitor the biomass growth of Aspergillus niger in solid-state fermentation (SSF) for lipase production using digital image processing technique. The strain A. niger 11T53A14 was cultivated in SSF using wheat bran as support, which was enriched with 0.91% (m/v) of ammonium sulfate. The addition of several vegetable oils (castor, soybean, olive, corn, and palm oils) was investigated to enhance lipase production. The maximum lipase activity was obtained using 2% (m/m) castor oil. In these conditions, the growth was evaluated each 24 h for 5 days by the glycosamine content analysis and digital image processing. Lipase activity was also determined. The results indicated that the digital image process technique can be used to monitor biomass growth in a SSF process and to correlate biomass growth and enzyme activity. In addition, the immobilized esterification lipase activity was determined for the butyl oleate synthesis, with and without 50% v/v hexane, resulting in 650 and 120 U/g, respectively. The enzyme was also used for transesterification of soybean oil and ethanol with maximum yield of 2.4%, after 30 min of reaction. PMID- 18401754 TI - The effects of surfactants on the estimation of bacterial density in petroleum samples. AB - The effect of the surfactants polyoxyethylene monostearate (Tween 60), polyoxyethylene monooleate (Tween 80), cetyl trimethyl ammonium bromide (CTAB), and sodium dodecyl sulfate (SDS) on the estimation of bacterial density (sulfate reducing bacteria [SRB] and general anaerobic bacteria [GAnB]) was examined in petroleum samples. Three different compositions of oil and water were selected to be representative of the real samples. The first one contained a high content of oil, the second one contained a medium content of oil, and the last one contained a low content of oil. The most probable number (MPN) was used to estimate the bacterial density. The results showed that the addition of surfactants did not improve the SRB quantification for the high or medium oil content in the petroleum samples. On other hand, Tween 60 and Tween 80 promoted a significant increase on the GAnB quantification at 0.01% or 0.03% m/v concentrations, respectively. CTAB increased SRB and GAnB estimation for the sample with a low oil content at 0.00005% and 0.0001% m/v, respectively. PMID- 18401755 TI - An alternative application to the Portuguese agro-industrial residue: wheat straw. AB - The effects of alkaline treatments of the wheat straw with sodium hydroxide were investigated. The optimal condition for extraction of hemicelluloses was found to be with 0.50 mol/l sodium hydroxide at 55 degrees C for 2 h. This resulted in the release of 17.3% of hemicellulose (% dry starting material), corresponding to the dissolution of 49.3% of the original hemicellulose. The yields were determined by gravimetric analysis and expressed as a proportion of the starting material. Chemical composition and physico-chemical properties of the samples of hemicelluloses were elucidated by a combination of sugar analyses, Fourier transform infrared (FTIR), and thermal analysis. The results showed that the treatments were very effective on the extraction of hemicelluloses from wheat straw and that the extraction intensity (expressed in terms of alkali concentration) had a great influence on the yield and chemical features of the hemicelluloses. The FTIR analysis revealed typical signal pattern for the hemicellulosic fraction in the 1,200-1,000 cm(-1) region. Bands between 1,166 and 1,000 cm(-1) are typical of xylans. PMID- 18401756 TI - The use of seaweed and sugarcane bagasse for the biological treatment of metal contaminated waters under sulfate-reducing conditions. AB - When wetlands reach maximum treatment capacity to remove heavy metals, removal can still take place through precipitation as sulfide because of the biological reduction of sulfate. To achieve this goal, anaerobic conditions must be attained, a sulfate source must exist, and an adequate substrate for sulfate reducing bacteria (SRB) is also required. In the present work, two ligneous cellulosic materials, a brown seaweed and sugarcane bagasse, have been selected as substrates for SRB growth. Experiments were simultaneously conducted in continuous operation in two columns (0.57 L each), one containing the ligneous cellulosic material plus inoculum and another containing only the ligneous cellulosic material. In this work, the removal of cadmium and zinc was studied because of their presence in effluents from mining/metallurgy operations. Results obtained indicated that the inoculated reactor was able to treat the effluent more efficiently than the noninoculated reactor considering the time course of the tests. PMID- 18401757 TI - Development of activity-based cost functions for cellulase, invertase, and other enzymes. AB - As enzyme chemistry plays an increasingly important role in the chemical industry, cost analysis of these enzymes becomes a necessity. In this paper, we examine the aspects that affect the cost of enzymes based upon enzyme activity. The basis for this study stems from a previously developed objective function that quantifies the tradeoffs in enzyme purification via the foam fractionation process (Cherry et al., Braz J Chem Eng 17:233-238, 2000). A generalized cost function is developed from our results that could be used to aid in both industrial and lab scale chemical processing. The generalized cost function shows several nonobvious results that could lead to significant savings. Additionally, the parameters involved in the operation and scaling up of enzyme processing could be optimized to minimize costs. We show that there are typically three regimes in the enzyme cost analysis function: the low activity prelinear region, the moderate activity linear region, and high activity power-law region. The overall form of the cost analysis function appears to robustly fit the power law form. PMID- 18401758 TI - Reaction kinetics of the hydrothermal treatment of lignin. AB - Lignins derived from abundant and renewable resources are nontoxic and extremely versatile in performance, qualities that have made them increasingly important in many industrial applications. We have shown recently that liquefaction of lignin extracted from aspen wood resulted in a 90% yield of liquid. In this paper, the hydrothermal treatment of five types of lignin and biomass residues was studied: Kraft pine lignin provided by MeadWestvaco, Kraft pine lignin from Sigma-Aldrich, organosolv lignin extracted from oat hull, the residues of mixed southern hardwoods, and switchgrass after hydrolysis. The yields were found dependent on the composition or structure of the raw materials, which may result from different pretreatment processes. We propose a kinetic model to describe the hydrothermal treatment of Kraft pine lignin and compare it with another model from the literature. The kinetic parameters of the presented model were estimated, including the reaction constants, the pre-exponential factor, and the activation energy of the Arrhenius equations. Results show that the presented model is well in agreement with the experiments. PMID- 18401759 TI - Hydrodynamic characterization of a column-type prototype bioreactor. AB - Agro-food industrial processes produce a large amount of residues, most of which are organic. One of the possible solutions for the treatment of these residues is anaerobic digestion in bioreactors. A novel 18-L bioreactor for treating waste water was designed based on pneumatic agitation and semispherical baffles. Flow patterns were visualized using the particle tracer technique. Circulation times were measured with the particle tracer and the thermal technique, while mixing times were measured using the thermal technique. Newtonian fluid and two non Newtonian fluids were used to simulate the operational conditions. The results showed that the change from Newtonian to non-Newtonian properties reduces mixed zones and increases circulation and mixing times. Circulation time was similar when evaluated with the thermal and the tracer particle methods. It was possible to predict dimensionless mixing time (theta (m)) using an equivalent Froude number (Fr (eq)). PMID- 18401760 TI - Thermal effects on hydrothermal biomass liquefaction. AB - Batch pressure vessels commonly used for hydrothermal liquefaction have typical heating times in the range of 30 to 60 min. Thermodynamically, the complex set of reactions are path dependent, so that the heating rate can possibly affect yields and the composition of the resultant liquid products. It is postulated that the mode of heat transfer becomes an uncontrolled variable in kinetic studies and can seriously impact scale-up. To confirm this hypothesis and minimize these heat transfer-related artifacts, we designed a batch pressure vessel equipped with an induction heating system, which allows the reduction of heat-up times by about two orders of magnitude to several seconds, compared to tens of minutes with standard pressure reactors. This system was used to study the direct liquefaction of corn stover and aspen wood with a pretreatment. The heating rate was found to have no significant effect on the composition of the liquid products. However, the liquid yields are dependent on the heating rate. Varying the cooling rate does not show obvious effects. The results confirm that the heating rate, as governed by the mode of heat transfer, is an important factor that needs to be considered during scale-up. PMID- 18401761 TI - Pattern of intra-family hetero-oligomerization involving the G-protein-coupled secretin receptor. AB - Oligomerization of G-protein-coupled receptors (GPCRs) is emerging as a mechanism for regulation and functional modification, although it has been studied most extensively for Family A receptors. Family B receptors have clear structural differences from Family A. In this paper, we have systematically evaluated GPCRs that are capable of association with the prototypic Family B secretin receptor. All of the receptor constructs were shown to traffic normally to the plasma membrane. We utilized receptor bioluminescence resonance energy transfer (BRET) to determine the presence of constitutive and ligand-dependent receptor association. Extensive intra-family and no cross-family association was observed. Of the nine Family B receptors studied, all constitutively yielded a significant BRET signal with the secretin receptor, except for the calcitonin receptor. Each of the associating hetero-oligomeric receptor pairs generated a BRET signal of similar intensity, less than that of homo-oligomeric secretin receptors. BRET signals from some receptor pairs were reduced by ligand occupation, but none were increased by this treatment. Thus, Family B GPCR oligomerization occurs, with many structurally related members associating with each other. The specific functional implications of this need to be further evaluated. PMID- 18401762 TI - A matter of identity: transcriptional control in oligodendrocytes. AB - Oligodendrocyte development progresses from specification to terminal differentiation through several phases. By now, a number of transcription factors have been identified that are essential for one or more of these phases. They stem from transcription factor families with known roles in many developmental processes. Basic helix-loop-helix, homeodomain, and high-mobility-group containing transcription factors such as the Olig, Nkx, and Sox proteins have been particularly well studied. A complex picture has emerged in which these transcription factors interact in transcriptional networks and thereby combine and influence their respective activities as repressors or activators in such a way that stage- and cell-type specific gene expression is achieved during oligodendrocyte development. PMID- 18401764 TI - High prevalence of hypovitaminosis D and secondary hyperparathyroidism in elders living in nonprofit homes in South Brazil. AB - OBJECTIVES: Hypovitaminosis D (HD) and secondary hyperparathyroidism (SHP) are common in elders, and many factors could contribute to them. The objectives of this study were to estimate the prevalence of HD, SHP, and its associated factors, in individuals living in nonprofit homes for elders in south Brazil. Design Cross-sectional study. METHODS: Serum 25-hydroxyvitamin D 25(OH)D, intact parathyroid hormone (PTH), total calcium, phosphorus, alkaline phosphatase, magnesium, creatinine, and albumin levels were measured in late spring, November, 2005. The presence of factors potentially related with HD and SHP-age, sex, weight, height, skin phototype, sun exposure, exercise, smoking, use of < or = 5 medications or diuretics or alcohol, and daily calcium ingestion. RESULTS: 102 subjects age 77.8 +/- 9.0 were included in the study. HD was found in 85.7% and SHP in 53% of the subjects. The estimated daily calcium ingestion was 720 mg. There was no association between serum 25(OH)D levels and any of the risk factors evaluated. Serum 25(OH)D levels were correlated with serum PTH (r = -0.358, P = 0.000), calcium (r = 0.306, P = 0.002), and albumin (r = 0.253, P = 0.011) levels. In univariate analysis, SHP was positively associated with age (P = 0.006), and female sex (0.007); and negatively associated with sunlight exposure (P = 0.020), GFR (P = 0.000), Ln25(OH)D (P = 0.002), and total serum calcium (P = 0.024). After multivariate model adjustment, age [OR 1.09 (CI 1.01-1.18); P = 0.024], Ln25(OH)D [OR 0.92 (CI 0.08-0.74); P = 0.013], GFR [OR 0.96 (CI 0.92 0.99); P = 0.013], and hydrochlorothiazide treatment [OR 7.63 (CI 1.67-34.9); P = 0.008] were independently associated with SHP. CONCLUSIONS: HD and SHP are highly prevalent in elders living in old-age homes. No associations were established between common risk factors and low serum levels of 25(OH)D levels; however, SHP was independently related with age, 25(OH)D, GFR, and hydrochlorothiazide use. PMID- 18401763 TI - Inhibition of rat granulosa cell differentiation by overexpression of Galphaq. AB - Activation of FSH and LH receptors in undifferentiated granulosa cells (i.e., no prior exposure to FSH) results in comparable induction of progesterone production, but activation of the LH receptor is less effective than FSH in inducing aromatase and the native LH receptor. Because the LH receptor can also activate the Galphaq signaling pathway, we investigated whether activation of this pathway could be responsible for these differences. Overexpression of Galphaq inhibited FSH induction of both the estradiol and progesterone biosynthetic pathways as well as mRNA levels for cholesterol side-chain cleavage enzyme (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), LH receptor (LHr), and P450aromatase (aromatase). This suppression was associated with a reduction (P < 0.05) in FSH-stimulated cAMP production. Lower cAMP levels were not due to reduced FSH receptor (FSHr) mRNA levels or reduced levels of Galphas. Phosphodiesterase (PDE) activity and regulator of G-protein signaling 2 (RGS2) mRNA levels were significantly (P < 0.05) increased by Galphaq, both of which could account for diminished cAMP levels. We conclude that Galphaq signaling pathway inhibits both estradiol and progesterone production comparably and thus activation of this pathway does not seem to account for differences between FSH and LH in the regulation of aromatase and the LH receptor. PMID- 18401765 TI - Effects of TGFbeta1 on gene expression in the HP75 human pituitary tumor cell line identified by gene expression profiling. AB - The pathogenesis of pituitary adenomas and many of the genes influencing growth of these tumors are unknown. TGFbeta is known to inhibit proliferation of cultured anterior pituitary cells and anterior pituitary tumors, but the signal transduction pathways involved in the inhibition of growth are unclear. We treated the human HP75 pituitary cell line with 10(-9) M TGFbeta1 for 4, 24, and 96 h and performed global gene expression profiling by Affymetrix GeneChip microarray analysis. Quantitative PCR validation of specific genes involved in the TGFbeta1-induced regulation of pituitary cell growth was also done. Of the 15,000 genes queried, there were 37 genes up-regulated and 48 genes down regulated twofold or more after 4 h of TGFbeta1 treatment. There were 121 genes up-regulated and 109 genes down-regulated twofold or more after 24 h of TGFbeta1 treatment and 112 genes up-regulated and 43 genes down-regulated twofold or more after 96 h of TGFbeta1 treatment. Galectin-3 (Gal-3) protein was decreased by TGFbeta1 treatment and several genes which interacted with Gal-3 including RUNX1 and WNT5B were up-regulated after TGFbeta1 treatment. SOX4 was also up-regulated by TGFbeta1 treatment. SMAD3, which is directly involved in the TGFbeta signal transduction pathway, was down-regulated by TGFbeta1 treatment. These findings highlight the diverse gene networks and pathways through which TGFbeta operates in its effects on pituitary tumor cells. PMID- 18401766 TI - Open-label, randomized, multiple-center, parallel study comparing glycemic responses and safety profiles of Glucerna versus Fresubin in subjects of type 2 diabetes mellitus. AB - Since the increase of prevalence of type 2 diabetes mellitus (T-2DM), the replacing quickly absorbed carbohydrates with a fat source rich in monounsaturated fatty acid to provide improved glycemic control in these patients has become an important assistant therapy. In the present study, we compared glycemic response and safety of two nutritional products, Glucerna and Fresubin, in Chinese subjects with T-2DM. Overall, 203 T-2DM subjects were randomly assigned (1:1) to either Glucerna or Fresubin. The primary endpoint was the adjusted area under the curve (adj-AUC) for plasma glucose at 0-240 min. Blood samples were collected at 0, 30, 60, 90, 120, 180, and 240 min to compare the adjusted area under the curve (AUC) for the change in plasma glucose or insulin from 0 to 240 min. Adjusted peak values and times of glucose and insulin responses and adjusted glucose and insulin values were collected at the same time points. Safety parameters were also evaluated. The adjusted AUC for the change in plasma glucose in the Glucerna group was significantly lower than in Fresubin group (5.60 +/- 5.88 mmol/l*h vs. 7.97 +/- 6.32 mmol/l*h, P = 0.0061), as was the adjusted peak value of glucose (3.51 +/- 2.04 mmol/l vs. 4.69 +/- 1.99 mmol/l, P < 0.0001). Glucerna subjects had a longer adjusted peak time to insulin response compared to Fresubin subjects (105.00 +/- 43.4 min vs. 88.81 +/- 37.69 min, P = 0.0050). Glucerna subjects also experienced more gradual changes in glucose and insulin values. In conclusion, Glucerna provided better control of postprandial plasma glucose and insulin levels in Chinese subjects with T-2DM. Variation of postprandial glucose tended to be relatively stable after patients took Glucerna. Study results suggest that Glucerna may be beneficial in the reduction of postprandial glycemia. PMID- 18401776 TI - Cellulose acetate microspheres as floating depot systems to increase gastric retention of antidiabetic drug: formulation, characterization and in vitro-in vivo evaluation. AB - Gastric emptying is a complex process that is highly variable and makes the in vivo performance of drug delivery systems uncertain. In order to avoid this variability, efforts have been made to increase the retention time of the drug delivery systems for more than 12 hours utilizing floating or hydrodynamically controlled drug delivery systems. The objective of this investigation was to develop a floating, depot-forming drug delivery system for an antidiabetic drug based on microparticulate technology to maintain constant plasma drug concentrations over a prolonged period of time for effective control of blood sugar levels. Formulations were optimized using cellulose acetate as the polymer and evaluated in vitro for physicochemical characteristics and drug release in phosphate buffered saline (pH 7.4), and evaluated in vivo in healthy male albino mice. The shape and the surface morphology of the prepared microspheres were characterized by optical microscopy and scanning electron microscopy. In vitro drug release studies were performed and drug release kinetics were calculated using the linear regression method. Effects of stirring rate during preparation and polymer concentration on the size of microspheres and drug release were observed. The prepared microspheres exhibited prolonged drug release (more than 10 hours) and remained buoyant for over 10 hours. Spherical and smooth-surfaced microspheres with encapsulation efficiency ranging from 73% to 98% were obtained. The release rate decreased and the mean particle size increased at higher polymer concentrations. Stirring speed affected the morphology of the microspheres. This investigation revealed that upon administration, the biocompatible depot-forming polymeric microspheres controlled the drug release and plasma sugar levels more efficiently than plain orally given drug. These formulations, with their reduced frequency of administration and better control over drug disposition, may provide an economic benefit to the user compared with products currently available for diabetes control. PMID- 18401777 TI - Antimicrobial activity of parabens in submicron emulsions stabilized with lecithin. AB - Antimicrobial efficacy of methyl and propylparaben combination as potential preservatives for submicron emulsions, and the effect of oil and lecithin concentration on the microbial growth were investigated. Parabens were ineffective in standard or doubled concentrations as per pharmacopoeial criteria. Poor growth inhibition and multiplication of reference strains point to protective and growth properties of submicron emulsions. No correlation was observed between oil/lecithin ratio and efficacy of parabens; partitioning of the latter into the oily phase and lipophilic domains could be the reason for such effect. Further studies are necessary to establish a stable and safe composition of such formulations. PMID- 18401767 TI - The Emerging Picture of Human Breast Cancer as a Stem Cell-based Disease. AB - There are increasing data supporting the existence of a cell hierarchy within the mammary gland. At the top or this hierarchy a small population of cells with self renewal properties maintains the tissue architecture and remodeling, and they are known as stem cells. Also, recent evidences indicate that breast cancer is originated and maintained by its own cancer stem cells reminding the normal mammary gland. The existence of this small population of cells with self-renewal capability has important biological and clinical significances. So, the interpretation of tumors as hierarchical cellular structures has changed our vision of the breast cancer scenario. Here, we review the current knowledge about normal and breast cancer stem cells, and their implications in cancer development, together with their consequences in breast cancer susceptibility, dissemination and treatment response. PMID- 18401778 TI - Optimization and evaluation of time-dependent tablets comprising an immediate and sustained release profile using artificial neural network. AB - The aim of this work was to optimize time-dependent tablets using artificial neural network (ANN). The time-dependent tablet consisted of a tablet core, which contained sustained release pellets (70% isosorbide-5-mononitrate [5-ISMN]), immediate release granules (30% 5-ISMN), superdisintegrating agent (sodium carboxymethylstarch, CMS-Na), and other excipients, surrounded by a coating layer composed of a water-insoluble ethylcellulose and a water-soluble channeling agent. The chosen independent variables, i.e., X(1) coating level of tablets, X(2) coating level of pellets, and X(3) CMS-Na level, were optimized with a three factor, three-level Box-Behnken design. Data were analyzed for modeling and optimizing the release profile using ANN. Response surface plots were used to relate the dependent and the independent variables. The optimized values for the factors X(1)-X(3) were 4.1, 14.1, and 29.8%, respectively. Optimized formulations were prepared according to the factor combinations dictated by ANN. In each case, the observed drug release data of the optimized formulations were close to the predicted release pattern. An in vitro model for predicting the effect of food on release behavior of optimized products was used in this study. It was concluded that neural network technique could be particularly suitable in the pharmaceutical technology of time-dependent dosage forms where systems were complex and nonlinear relationships often existed between the independent and the dependent variables. PMID- 18401779 TI - A novel superdisintegrating agent made from physically modified chitosan with silicon dioxide. AB - Disintegrants and fillers represent important excipients for immediate-release solid dosage forms in many pharmaceutical applications. A new excipient based on the coprecipitation of chitosan and silica has been achieved. The "intimate" physical association between chitosan and silica creates an insoluble, hydrophilic, highly absorbent material, consequently, resulting in superiority in water uptake, water saturation for gelling formation, and compactability among other superdisintegrants. The new excipient has an outstanding functionality that does not primarily depend on water wicking and swelling properties. In fact, it translates it into superior disintegration characteristics with improved powder flow and compaction properties. Thus, the new excipient could act as a superdisintegrant and pharmaceutical filler at the same time. Studies have shown that chitosan-silica delivers superior performance in wet granulation formulations and is the only disintegrant that is effective at all concentrations in tablet formulation. PMID- 18401780 TI - Basil oil is a promising skin penetration enhancer for transdermal delivery of labetolol hydrochloride. AB - The present work investigates effectiveness of basil oil, a volatile oil containing alcoholic terpenes, as a potential penetration enhancer for improved skin permeation of labetolol hydrochloride (LHCl) with reference to camphor, geraniol, thymol, and clove oil. Saturation solubilities of LHCl were determined in water, vehicle (ethanol:water, 60:40 v/v) and vehicle containing 5% w/v terpenes. Comparable (P > 0.05) saturation solubilities were found suggesting an insignificant increase in LHCl flux across rat skin on account of thermodynamic activity. Permeation of LHCl in vehicle per se and in presence of 5% w/v enhancer was investigated by performing in vitro rat abdominal skin permeation studies using a side-by-side glass diffusion cell. Various parameters viz. steady state flux, permeability coefficient, lag time, partition coefficient, diffusion coefficient, and enhancement ratios (ER) were calculated from the permeation data. Basil oil produced the maximum enhancement (ER = 46.52) over neat vehicle, among all enhancers. Activation energies for LHCl permeation in water, vehicle per se and in presence of 5% w/v basil oil were found to be 23.16, 18.71, and 10.98 kcal/mole, respectively. Lowering of activation energy in presence of basil oil suggests creation of new polar pathways in the skin for enhanced permeation of LHCl. Basil oil is proposed as a promising penetration enhancer for improved transdermal drug delivery of labetolol. PMID- 18401781 TI - Mechanistic analysis of drug release from theophylline pellets coated by films containing pectin, chitosan and Eudragit RS. AB - The objective of this study was to obtain detailed information on the mechanism of drug release from mixed-film of pectin-chitosan/Eudragit RS. Pellets (710-840 microm in diameter) containing 60% theophylline and 40% microcrystalline cellulose were prepared by extrusion-spheronization method. Eudragit L100-55 enteric coating capsules included film-coated pellets of theophylline in theoretical coating weight gains of 10, 15, and 20%, with pectin-chitosan complex contents of 5, 10, 15, and 20% for each level of weight gain were prepared and subjected to in vitro drug release. Drug release from this system showed a bimodal release profile characteristic with the drug release enhancement, being triggered (burst release) in the colonic medium. The reason for burst drug release may be due to the enzymatic degradation of pectin via pectinolytic enzymes in the simulated colonic medium. The mechanism of drug release from each formulation was evaluated in the terms of zero-order, first-order, Higuchi and Korsmeyer-Peppas models. It was observed that none of the enteric coating capsules showed any drug release in the simulated gastric medium (phase I). The analysis of release profiles showed that zero-order kinetics was found as the better fitting model for all formulations in the simulated small intestine (phase II) and it could be due to the pectin-chitosan swelling and subsequent formation of aqueous channels. In the colonic medium (phase III), due to degradation of pectin and its leaching from the mixed-film, there was a modification in drug release kinetics from swelling-controlled at phase II to anomalous at phase III. It also was found that both zero-order and Higuchi models contributed in colonic drug release from most of the formulations. PMID- 18401782 TI - Enhanced percutaneous permeability of diclofenac using a new U-type dilutable microemulsion. AB - Enhanced systemic absorption in vivo and percutaneous penetration in vitro was demonstrated after transdermal administration of diclofenac sodium formulated in U-type microemulsion. Diclofenac sodium was solubilized in a typical four component system consisting of an oil, polyoxyethylene-10EO-oleyl alcohol (Brij 96V) as the surfactant, and 1-hexanol along water dilution line W46 (40 wt % surfactant and 60 wt % oil phase before water titration). Viscosity and small angle X-ray scattering measurements have evidenced bicontinuous structures within water fractions of 0.25 and 0.5 along the dilution line. Self-diffusion NMR studies showed that drug molecules accumulated in the interfacial film and, to some extent, dissolved in the oil. Relative to a commercial macro-emulsion cream (Voltaren Emulgel), microemulsions containing paraffin oil or isopropyl myristate increased the in vivo transdermal penetration rate of diclofenac by two order of magnitude, whereas the rat plasma levels were increased by one order of magnitude. The in vitro data obtained from excised rat skin were comparable to the in vivo results, but suffered from discrepancies from the ideal in vivo-in vitro correlation, which might be explained by optimal in vitro conditions of perfusion and hydration. It has also been found that when jojoba oil is formulated as the oil phase in the microemulsion, the penetration rate of the drug decreases significantly. Based on the three-dimensional structure of jojoba oil, the wax is presumed to prevent the drug from being freely diffused into the skin while migrating from the interfacial film into the continuous oil phase. PMID- 18401783 TI - A thermo-sensitive polymeric gel containing a gadolinium (Gd) compound encapsulated into liposomes significantly extended the retention of the Gd in tumors. AB - Gadolinium neutron capture therapy (Gd-NCT) is a promising approach to fight cancer. One key factor for the success of Gd-NCT is to deliver and maintain a sufficient amount of Gd inside tumors. A large amount of Gd can be readily introduced into tumors by direct intratumor injection. However, an innovative approach is needed to maintain the Gd in the tumors. We encapsulated a Gd compound into a liposome formulation and then dispersed the liposomes into a thermo-sensitive polymeric gel. In murine tumor models, we showed that this liposome-in-thermo-sensitive gel system significantly extended the retention of the Gd compound in tumors. This similar concept may be applied to prolong the retention of other cytotoxic chemicals in tumors, and thus, improve their anti tumor efficacy. PMID- 18401784 TI - Encapsulation of indomethacin using coaxial ultrasonic atomization followed by solvent evaporation. AB - We have encapsulated indomethacin into poly (lactide-co-glycolide) (PLGA) using coaxial ultrasonic atomization technique. The specific aims of this study were to evaluate the effect of drug loading and a change in relative concentration of polymer in the inner and outer layers of coflowing spray liquids on the physicochemical characteristics of the particles. Indomethacin, a non steroidal anti-inflammatory drug, was selected as a model compound. The micro/nanocapsules prepared using a drug free PLGA solution as an outer layer showed higher encapsulation efficiency. Thermal analysis of the formulations indicated that indomethacin was dissolved within the PLGA matrix. The formulations prepared with 25 mg indomethacin showed relatively smaller particle size compared with the formulations prepared with 50 mg indomethacin. The particles, in general, showed bi- and tri-modal distribution. Irrespective of the compositions of the liquids 1 and 2, all the particles were smooth and spherical. A cross-section view of the particles revealed the presence of three different internal morphologies. These formulations were a mixture of hollow or solid spheres, and single or multiple spheres encapsulated into a larger sphere. To the best of our knowledge, this is the first study revealing the cross-sectional view of particles prepared with coaxial ultrasonic atomization technique. PMID- 18401785 TI - Freeze drying of double emulsions to prepare topotecan-entrapping liposomes featuring controlled release. AB - Topotecan-entrapping liposomes were prepared by freeze drying double emulsions with hydrogenated soy phosphatidylcholine, N-(carbonyl methoxypolyethyleneglycol2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and cholesterol. Different inner aqueous phases of different pH values containing topotecan together with different chemicals, such as citrate and sulfate, were used to modify the physicochemical state of the drug to prepare W1/O/W2 double emulsions that were then freeze dried to obtain dry products. Upon rehydration, the dry products, which were stable for at least 6 months, formed into unilamellar liposomes with a high encapsulation efficiency of up to 80% and a mean diameter below 200 nm. The in vitro release experiments demonstrated that different formulations displayed different drug release properties. Thus, stable submicron unilamellar topotecan-entrapping liposomes can be prepared by freeze drying double emulsions, and the drug release can be successfully controlled by altering the physicochemical state of the incorporated drug. PMID- 18401786 TI - Diclofenac-bismuth complex: synthesis, physicochemical, and biological evaluation. AB - Diclofenac-bismuth complexation was attempted by mixing diclofenac sodium (Na) and bismuth-subcitrate aqueous solutions at diclofenac:bismuth molar ratio of 3:1. A solid precipitate was obtained and isolated. The precipitate was characterized for stoichiometric ratio of diclofenac-bismuth complexation using capillary electrophoresis, which showed 1:1 complexation. In addition, nuclear magnetic resonance and Fourier transform infrared analysis were performed for the isolated solid complex and indicated that bismuth was in coordinate bond formation with the carboxylate group of diclofenac. In comparison with diclofenac Na powder, the complex was evaluated as an aqueous suspension for in vitro drug dissolution. The complex exhibited a faster dissolution rate than and similar dissolution extent as diclofenac Na. In comparison with an aqueous solution of diclofenac Na and an aqueous suspension of physical mixture of diclofenac acid (suspended) and bismuth-subcitrate (dissolved), the aqueous complex suspension was evaluated for ulcerogenic effect in rats upon oral administration. The complex led to more gastric ulceration than diclofenac Na, which was not in accordance with the antiulcer properties of bismuth. This antiulcer effect was shown as the physical mixture administration was accompanied with lower gastric ulceration than diclofenac Na administration. These gastric ulceration results were explained in terms of the difference in particle size between solid diclofenac acid formed as a result of the complex breakdown in an acidic medium (0.1 M HCl to simulate the gastric fluid) and that formed as a result of diclofenac Na neutralization. Diclofenac acid particles formed from the complex breakdown were of average size, three times smaller of those formed as a result of diclofenac Na protonation. This difference in particle size was correlated with the higher gastric ulceration associated with the complex than with diclofenac Na in terms of higher coverage of the gastric mucosa with diclofenac, and consequently, higher local ulceration. PMID- 18401787 TI - In situ ophthalmic gel of ciprofloxacin hydrochloride for once a day sustained delivery. AB - This article focuses on preparation and evaluation of a once a day ophthalmic delivery system for ciprofloxacin hydrochloride based on the concept of pH triggered in situ gelation. The in situ gelling system involves the use of polyacrylic acid (Carbopol 980NF) as a phase transition polymer, hydroxypropyl methylcellulose (Methocel K100LV) as a release retardant, and ion exchange resin as a complexing agent. Ciprofloxacin hydrochloride was complexed with ion exchange resin to avoid incompatibility between drug and polyacrylic acid. The developed formulation was stable, and nonirritant to rabbit eyes and in vitro drug release was found to be around 98% over a period of 24 hours. PMID- 18401788 TI - Investigation of nanostructured lipid carriers for transdermal delivery of flurbiprofen. AB - The aim of this study was to develop nanostructured lipid carriers (NLC) for transdermal delivery of Flurbiprofen (FP). The physical stability of FP-NLC and its in vitro permeation profile were investigated. After three months of storage at 4 degrees C, 20 degrees C, and 40 degrees C, no significant differences between the evaluated parameters, such as pH value, the entrapment efficiency, particle size, and zeta potential were observed. In in vitro permeation studies, the cumulative permeated amounts and the release rate from FP-NLC were 412.53 +/- 21.37 microg/cm(2) and 35.25 microg/cm(2)/h after 12 h (n = 6), respectively, while from saturated FP-PBS (pH = 7.4) were 90.83 +/- 8.67 microg/cm(2) and 6.99 microg/cm(2)/h, respectively. These results indicated that the FP-NLC were with good physical stability and were able to improve the permeated amounts and the release rate of FP. It could potentially be exploited as a carrier with improved drug entrapment efficiency and permeated amount in the transdermal delivery of FP. PMID- 18401789 TI - Measurement of the physical properties of aerosols in a fullerene factory for inhalation exposure assessment. AB - Assessment of human exposure is important for the elucidation of potential health risks. However, there is little information available on particle number concentrations and number size distributions, including those of nanoparticles, in the working environments of factories producing engineered nanomaterials. The authors used a scanning mobility particle sizer and an optical particle counter to measure the particle number size distributions of particles ranging in diameter (D(p)) from 10 nm to >5000 nm in a fullerene factory and used scanning electron microscopy to examine the morphology of the particles. Comparisons of particle size distributions and morphology during non-work periods, during work periods, during an agitation process, and in the nearby outdoor air were conducted to identify the sources of the particles and to determine their physical properties. A modal diameter of 25 nm was found in the working area during the non-work period; this result was probably influenced by ingress of outdoor air. During the removal of fullerenes from a storage tank for bagging and/or weighing, the particle number concentration at D(p)<50 nm was no greater than that in the non-work period, but the concentration at D(p)>1000 nm was greater during the non-work period. When a vacuum cleaner was in use, the particle number concentration at D(p)<50 nm was greater than that during the non work period, but the concentration at D(p)>1000 nm was no greater. Scanning electron microscopy revealed that the coarse particles emitted during bagging and/or weighing were aggregates/agglomerates of fullerenes; although origin of particles with D(p)<50 nm is unclear. PMID- 18401791 TI - [Basic principles and clinical application of retinal laser therapy]. AB - The scientific background of laser photocoagulation of the ocular fundus was studied extensively by several investigators in the 1970 s and 1980 s. The basic principles were successfully resolved during that time and clinical consequences for proper application of the laser photocoagulation for various diseases were deduced. The present paper gives an overview about the physical basics of laser tissue interactions during and after retinal laser treatment and the particular laser strategies in the treatment of different retinal diseases. Thus, it addresses the issue of the impact on tissue of laser parameters as wavelength, spot size, pulse duration and laser power. Additionally, the different biological tissue reactions after laser treatment are presented, such as, e. g., for retinopexia or macular treatments as well as for diabetic retinopathies. Specific laser strategies such as the selective laser treatment of the RPE (SRT) or the transpupillary thermotherapy (TTT) are presented and discussed. PMID- 18401792 TI - [The retinal organ culture--a model system for the examination of the early cytoskeletal reaction pattern after retinal detachment]. AB - BACKGROUND: In vivo animal experiments have shown that the cytoskeleton plays a crucial role in case of structural changes after an induced retinal detachment. This study attempts to clarify whether a retinal organ culture could serve as an in vitro model for retinal detachment and thus represent an alternative to animal experiments. The main focus of this publication lies on the early cytoskeletal changes after retinal detachment. MATERIALS AND METHODS: Porcine retinas were mounted on special carriers, cultured for one or two weeks and examined by standard immunohistological (vimentin, GFAP, alpha-tubulin), as well as electron microscopical procedures. RESULTS: The cytoskeletal changes revealed similar spatio-temporal pattern compared with in vivo induced retinal detachments. In addition, it was shown that microtubules might play a crucial role in the early phase of gliosis, i. e., prior to a subretinal invasion by Muller cell extensions. CONCLUSIONS: The presented organ culture model will be used to unravel the largely unknown initial reactions of retinal gliosis, focusing on subcellular changes localised at the outer limiting membrane. The intracellular transport system of microtubules might play a key role in these processes. PMID- 18401793 TI - [Intraepithelial phototherapeutic keratectomy and alcohol delamination for recurrent corneal erosions--two minimally invasive surgical alternatives]. AB - BACKGROUND: Phototherapeutic keratectomy (PTK) has become established as a successful therapy for recurrent corneal erosions. After epithelial debridement, Bowman's lamella and anterior stroma are ablated by the Excimer laser. We have evaluated two alternative stroma-sparing treatment options, intraepithelial PTK, and alcohol delamination. PATIENTS AND METHODS: All treatments were performed in the relapse-free period. 17 eyes with recurrent corneal erosions were treated with intraepithelial PTK: from the intact epithelium, 12 - 25 microm of tissue were ablated by the Excimer laser (group I). Alcohol delamination was performed in 13 eyes (group II). Follow-up time was between 6 months and 7 years (mean 4.2 years). RESULTS: Both methods turned out to be safe, no refractive changes were detectable. After intraepithelial PTK, we saw a cumulative recurrence rate of 12 % after 1 year, 18 % after 2 years, and 24 % after 3 years, and a temporary subepithelial scaring was seen. Alcohol delamination resulted in a recurrence rate of 15 % during the whole follow-up time (no statistically significant difference compared to intraepithelial PTK), showing no haze or scarring. CONCLUSION: Both minimally invasive, stroma-sparing methods were effective for the treatment of trauma-associated recurrent erosion. The ablation of Bowman's lamella or anterior stroma does not seem to be necessary. However, for basal membrane dystrophy, we recommend PTK after epithelial debridement for the partial ablation of Bowman's lamella. PMID- 18401794 TI - [Atypical macular holes]. AB - BACKGROUND: Therapeutic strategies for macular holes have been optimised during the last years. However, little is known about atypical macular holes. This study was conducted to analyse the clinical and anatomic outcome in secondary macular holes of different origins. PATIENTS AND METHODS: In a retropective analysis 60 eyes with atypical macular holes that underwent surgical repair were identified. Demografic data, lens status, macular situation, best corrected visual acuity (BCVA) pre- and postoperative and complications were documented. After exclusion of patients with trauma, vitreomacular traction syndrome and epiretinal gliosis four subgroups were analysed. Group I: after retinal detachment (n = 6), Group II: with retinal vein occlusion (n = 5), Group III: associated with diabetic macular oedema (n = 6), Group IV: during/after internal limiting membrane peeling (n = 3). RESULTS: I. Four of six eyes showed a macular hole after successful retinal detachment surgery and two eyes in the presence of retinal detachment. Five of six eyes showed postoperative closure of the macular hole. BCVA improved in four eyes, worsened in one eye and remained unchanged in one. II. In five eyes a secondary macular hole occurred after retinal vein occlusion. After vitrectomy and gas tamponade a successful hole repair was observed in all eyes. Improvement of BCVA occurred in four eyes and remained unchanged in one eye. III. In six eyes a secondary macular hole developed after rupture of cysts in diabetic macular oedema. Four of six holes were closed successfully after vitrectomy. Improvement of BCVA was seen in two eyes, impaired BCVA in one eye and in one eye vision remained unchanged. IV. This group consists of two eyes with a macular hole after vitrectomy and membrane peeling and one eye with an iatrogenic intraoperatively created macular hole. After vitrectomy and gas tamponade, anatomic success was achieved in two eyes. Improvement of BCVA was observed in two eyes, in one eye BCVA deteriorated markedly. CONCLUSIONS: In spite of the different underlying diseases and pathomechanisms, secondary macular holes can be treated successfully in the majority of cases. Visual recovery was moderate in patients with diabetic macular oedema but marked in the other subgroups. Therefore, vitrectomy seems reasonable also in non-atypical macular holes of various origins. PMID- 18401795 TI - [Autologous translocation of large retinal pigment epithelium choroid patches in age-related macular degeneration]. AB - BACKGROUND: The latest development in ARMD surgery is the translocation of an autologeous pigment epithelium choroid patch. The method has technical shortcomings: The transplant is excised including the overlaying retina and inserted through a retinotomy near the posterior pole thus causing iatrogenic field defects. For the same reasons the size of the transplant is limited. MATERIALS AND METHODS: The technique was modified as follows: lens surgery using a special PCL with equal power in water and silicone oil, 180 degrees retinotomy just at the temporal ora serrata, subretinal surgery including patch transplantation with the retina folded over nasally and fixed by PFCL, complete silicone oil tamponade without any water remaining. PATIENTS: 12 consecutive cases, age 79 (70 - 86) years, 4 RPE detachments and rips, 8 subretinal hemorrhages from wet ARMD, follow-up in 10 eyes over 15.3 (3 - 23) months. Time courses for visual acuity, depth of central scotoma, OCT and FAG. RESULTS: The mean diameter of the transplants was 16.5 (9 - 33) degrees . Silicone explantation in 7 / 10. Complications in 3 / 10: 1 macula puckering, 1 peripheral detachment, 1 PVR detachment. According to FAG the transplant vascularises in 4 - 6 weeks. 4 / 10 eyes reached visual acuity > 0.2 with limited reading capability. Central scotoma depth remained constant at -11 dB. Function deteriorated again after 6 - 9 months with cystoid degeneration and retinal thickening. 8 / 11 patients estimated the operated eye to be superior to the untreated partner eye. CONCLUSIONS: Patch transplantation is able to restore limited reading capability in eyes having minor damage of the central retina. The best cases for this type of operation are RPE rips and recent sub-RPE haemorrhages. The functional success lasts 5 to 9 months, then the retina over the transplant begins to degenerate. PMID- 18401796 TI - [Intravitreal anti-VEGF therapy for capillary hemangioblastomas in von Hippel Lindau disease]. AB - BACKGROUND: Up-regulated expression of the vascular endothelial growth factor (VEGF) in von Hippel-Lindau (VHL) disease has been postulated to induce retinal hemangioblastoma. Intravitreal injections of anti-VEGF drugs might provide a new therapeutic option in this condition. METHODS: In a single case decision a patient with active retinal hemangioblastomas due to VHL disease received repeated intravitreal injections of 0.5 mg ranibizumab. RESULTS: Subsequent to intravitreal anti-VEGF therapy, the signs of activity of the retinal hemangioblastomas slowly regressed. CONCLUSIONS: Intravitreal anti-VEGF therapy might, as monotherapy or as combination therapy, offer a new treatment option for retinal hemangioblastoma. PMID- 18401797 TI - [Solitary albinotic spot of the retinal pigment epithelium: a functional and imaging study]. AB - PURPOSE: The presence or absence of functional changes associated with solitary, congenital, hypopigmented lesions of the retinal pigment epithelium (RPE) have been a matter of controversy. This case report describes retinal and functional findings in a young patient with such a lesion. METHODS: A 10-year-old Hispanic female with a solitary congential hypopigmented spot of the RPE was examined using fundus photography, fluorescein angiography, autofluorescence imaging (AF) and optical coherence tomography (OCT). Functional analyses were performed using the Humphrey 24 - 2 visual field, Goldmann perimetry and the multifocal ERG (mfERG). RESULTS: A small visual field defect was demonstrated on both Goldmann perimetry (I/ 2e test object) and on Humphrey 24 - 2 visual field testing (significant at the 0.5 % level for pattern deviation). The multifocal ERG response amplitudes were decreased in the corresponding area and increased in implicit time. Autofluorescence imaging showed an absence of fluorescence corresponding to the area of the lesion. OCT findings were indicative of a small amount of subretinal fluid or schisis-like changes overlying the RPE anomaly. CONCLUSION: The results indicate that solitary, albinotic spots of the RPE can be associated with visual field defects and outer retinal deficits; these may be related to impaired RPE function and/or chronic exudative changes. PMID- 18401798 TI - [40 years of incomplete silicone oil tamponade after retinal detachment surgery]. AB - INTRODUCTION: Since the early 1970 s, silicone oil injection was used for the treatment of complicated retinal detachments. Especially in this early period, the possible complications of silicone oil tamponade were difficult to assess. Today, we strive to reduce the duration of silicone oil tamponade - aiming at a removal within 3 - 6 months. A permanent tamponade is reserved for special situations. PATIENT AND METHOD: We report on a 53-year-old male patient, who underwent retinal surgery and was followed-up for 40 years. RESULTS: After a blunt trauma at the age of 13 the patient developed a macula-off retinal detachment. An encircling procedure (Arruga suture) with combined incomplete silicone oil injection was performed. Best corrected visual acuity (BCVA) preoperatively was 20 / 60 and 20 / 100 was attained postoperatively. As a complication of silicone oil tamponade a localised cataract formation was observed. At the last visit BCVA was 20 / 200. No other silicone oil-related complications were noticed. CONCLUSION: Long-term silicone oil tamponade over a considerable period of 40 years was well tolerated without significant complications. The incomplete silicone oil filling of the vitreous cavity may be responsible for the lack of serious side effects and segmental cataract. PMID- 18401799 TI - [Leber's hereditary optic neuropathy on initial suspicion of optic neuritis]. PMID- 18401800 TI - [Medicinal herbs in ophthalmology in the manuscript Macer floridus and a comparison with the de materia medica of dioskurides, the codex medicina antiqua and the Viennese dioscurides]. PMID- 18401801 TI - A new squash specific incremental field test. AB - The purpose of this study was to develop a practical and reliable squash specific incremental test for estimating maximal oxygen uptake (VO(2max)). VO(2max) was measured using breath-by-breath online gas analysis among 11 male and 10 female squash players during a maximal incremental running treadmill protocol. All subjects performed a specially designed maximal squash specific incremental test (SSIT) until fatigue on two separate occasions. There were positive intra-class correlations for the two SSIT trials for time to fatigue (r=0.998, p<0.001) but the second trial was shorter (t=2.7, p<0.05) due to less stable scores among female players. A positive correlation was found between SSIT performance and VO(2max) (r=0.924, p<0.001). SSIT time to fatigue and VO(2max) regression equations were used to produce a table to predict VO(2max) from SSIT performance. SSIT was found to be a reliable and valid method of estimating VO(2max) among squash players. A balance between usability and squash specificity was achieved in the design of the SSIT without jeopardising its purpose of estimating VO(2max). SSIT is inexpensive, quick to set up and simple to administer compared to other squash specific tests and can be used with novice through to elite players. PMID- 18401802 TI - The role of resting duration in the kinematic pattern of two consecutive bench press sets to failure in elite sprint kayakers. AB - This study aimed to investigate the role of rest period duration (RP) on the time course of the acceleration portion (AP) and mean velocity of the concentric phase across two bench press sets to failure with a submaximal load (60% of the 1RM) using different RP. Ten elite junior kayakers performed, on four different days, two consecutive bench press sets to failure, allowing randomly 1-, 2-, 3- and 4 min RP between sets. AP reached a maximal value of 66% of the concentric movement time. This maximal AP was observed in repetition number 2 or 3, and then AP declined during the set, with a significant decrease when the number of repetitions was over 80% of the total number of repetitions performed. AP and lifting velocity patterns of the concentric phase were not altered during a second set to failure, regardless of RP. However, when velocity was expressed in absolute terms, 1-min RP was insufficient to maintain the average lifting velocity during the second set, compared to the first one. These results may be of use in selecting number of repetitions and resting duration in order to ensure optimal maintenance of the accelerative portion of concentric movement time with different resting-period durations. PMID- 18401803 TI - Auxiliary muscles and slow component during rowing. AB - The aim of this study was to investigate the contribution of the auxiliary muscles, utilized to sustain the subject's position on the ergometer, to the oxygen uptake slow component phenomenon. Three tests were performed at the same severe relative intensity on a rowing ergometer: a standard rowing exercise test, a rowing exercise performed with the arms and one performed with the legs only. During the three exercise modalities, oxygen uptake, local oxyhemoglobin saturation and surface electromyography signals of the trapezius and vastus lateralis muscles were measured. The slow component amplitude, in absolute values, resulted statistically lower for rowing (343.9 ml . min (-1)) than for arms (795.6 ml . min (-1)) and legs (695.8 ml . min (-1)) exercise modes. The same result was found when the slow component amplitude was calculated as percentage of V O (2peak) (7.1 % for rowing; 17.2 % for arms; 17.3 % for legs). The lower slow component amplitude measured for the rowing exercise mode with respect to both arms and legs modes, demonstrates that the auxiliary muscles involved in the exercise contribute to the increasing energetic cost due to the slow component. PMID- 18401804 TI - Validity of the running repeated sprint ability test among playing positions and level of competitiveness in trained soccer players. AB - The purpose of this study was to examine the construct validity of the running repeated sprint ability (rRSA) test to discriminate performances between: i) various playing positions (Study 1), and ii) teams of different level of competitiveness (Study 2), in trained soccer players. Study 1 comprised clubs' players participating in the top local professional league. Study 2 comprised professional and semiprofessional players from the National Under-23 and Youth squads respectively, and amateur-level players from a University team. The rRSA test protocol consisted of either 6 or 8 repetitions of 20-m all-out sprints, interspersed with a 20-s active recovery period. There were significant differences in the rRSA performance between goalkeepers and outfield-positions (p < 0.01). Forwards had significantly better rRSA performance compared to defenders and midfielders (F ratio = 4.147, p = 0.02). Performance in the rRSA was superior in teams with relatively higher competitiveness (F ratio = 3.973, p = 0.02). The rRSA is a specific physical or fitness attribute of an outfield-position player, and may be of greater importance to the forwards' position. Performance in the rRSA also seems to be associated with a higher level of competitiveness and/or adaptation to resistance training. These data support the construct validity of the rRSA test in trained soccer players. PMID- 18401805 TI - Skeletal muscle pathways of contraction-enhanced glucose uptake. AB - Muscle contraction acutely increases glucose transport in both healthy and type 2 diabetic individuals. Since glucose uptake during muscle contraction has been observed in the absence of insulin, the existence of an insulin-independent pathway has been suggested to explain this phenomenon. However, the exact mechanism behind the translocation of GLUT4 vesicles through the sarcolemma during muscle contraction is still unknown. Some substances, such as AMPK and calcium activated proteins, have been suggested as potential mediators but the exact mechanisms of their involvement remain to be elucidated. A hypothetical convergence point between the insulin cascade and the potential pathways triggered by muscle contraction has been suggested. Therefore, the earliest concept that two different routes exist in skeletal muscle has been progressively modified to the notion that glucose uptake is induced by muscle contraction via components of the insulin pathway. With further consideration, increased glucose uptake and enhanced insulin sensitivity observed during/after exercise might be explained by a metabolic- and calcium-dependent activation of several intermediate molecules of the insulin cascade. This paper aimed to review the literature in order to examine in detail these concepts behind muscle contraction induced glucose uptake. PMID- 18401806 TI - Testosterone and BMD in elite male lightweight rowers. AB - The purpose of the present study was to investigate if a relationship between BMD and testosterone levels could be identified in elite male lightweight rowers. Thirteen male lightweight national team rowers had their BMD measured in a DEXA scanner. Plasma concentrations of total testosterone (TT), free testosterone (FT), dihydrotestosterone (DHT) and sex hormone binding globulin (SHBG) and additional parameters related to bone metabolism were measured. Plasma concentrations of TT, FT and DHT were in the lower part of the normal range, while BMD was close to or above normal. BMD of total body and L2 - L4 were correlated to years of training (r (s): 0.59, p = 0.034 and r (s): 0.73, p = 0.005) and to TT (r (s): 0.56, p = 0.046 and rs: 0.63, p = 0.021). Moreover, L2 - L4 BMD was correlated to FT (r (s): 0.62, p = 0.024). After adjusting for years of training, partial correlation analysis showed a significant correlation between L2 - L4 BMD and TT (r (s): 0.61, p < 0.05). BMD appears to be influenced by both testosterone levels and years of training in elite male lightweight rowers. The relatively high BMD and low testosterone levels indicate that the mechanical loading induced by rowing is more important to BMD than testosterone levels. Prospective investigations are needed to elucidate potential causal relationships. PMID- 18401807 TI - Anthropometric characteristics of ultramarathoners. AB - Little is known about the anthropometric characteristics of ultramarathon runners. The present work reports on the physical characteristics of the 392 (310 males, 82 females) starters and 270 (216 males, 54 females) finishers of the 2007 Western States Endurance Run, one of the largest 161-km trail runs in North America. Among the starters, mean (and 25th to 75th percentiles) body mass index (BMI) values were 23.2 (21.6 - 24.6) and 20.6 (19.4 - 21.9) kg . m (-2) for the men and women, respectively. Men were significantly taller, heavier and had greater BMI's across all age groups compared with the women. Among the top-5 overall finishers, mean BMI values were 23.2 (range 22.4 - 24.7) for the men and 19.8 (range 17.3 - 21.1) for the women. Average running speed and BMI were negatively correlated for both men (r (2) = 0.11, p < 0.0001) and women (r (2) = 0.10, p = 0.02). From this analysis, it is concluded that those participating in ultramarathon runs can vary widely in physical characteristics with BMI values that would classify some individuals as underweight and others as overweight. BMI varied considerably even among the top finishers, but lower BMI values were associated with faster running times. PMID- 18401808 TI - Testosterone and Ca2+ regulation in skeletal muscle. AB - The aim of this study was to examine the effect of testosterone treatment on the expression of dihydropyridine and ryanodine receptors in skeletal muscle of mouse. Furthermore, the effects of training, a method also known to elevate the plasma testosterone level, were studied and compared to the effects of pure testosterone administration. Male mice were either administered with testosterone or trained with treadmill. After 6 weeks, hindlimb muscles were excised and the expression of receptors was measured by Western blotting. Furthermore, the alterations in myosin heavy chain phenotypes were studied. In general, both training and testosterone administration induced changes in the expression of both receptors and in myosin heavy chain composition. In testosterone treated mice the expression of dihydropyridine receptor in extensor digitorum longus was higher compared to the control ones (38.9 %, p = 0.026). In soleus the expression was quite the contrary (- 27.3 %, p = 0.044), as was the case with ryanodine receptor (- 51.4 %, p = 0.012). The amount of ryanodine receptors was higher in rectus femoris (144.0 %, p = 0.044) and plantaris (48.1 %, p = 0.037) in testosterone treated mice. In trained mice, the expression of ryanodine receptor was significantly higher in gastrocnemius (27.6 %, p = 0.018), soleus (57.2 %, p = 0.025), plantaris (28.5 %, p = 0.009) and extensor digitorum longus (94.8 %, p = 0.009) than in the control ones. No differences were observed in the dihydropyridine receptor level. To conclude, training has a more important role in skeletal muscle adaptation compared to increased plasma testosterone level. However, in postural muscles both treatments have comparable effects. PMID- 18401809 TI - Effect of a whole-body vibration session on knee stability. AB - The aim of the study was to investigate the effect of WBV on stretch reflexes involved in knee joint control. We evoked stretch reflexes of the hamstring muscles by inducing an anterior tibial translation during standing in 23 healthy subjects which were divided into a control and an intervention group. WBV with a frequency of 30 Hz and a vertical amplitude of 4 mm was induced by an uniformly oscillating platform. The WBV session lasted 60 seconds and was repeated twice. Short (SLR) and medium latency responses (MLR) of the hamstring muscles and maximum tibia translation were assessed using surface EMG and linear potentiometers. While there were no significant changes in latency, the size of the lateral and medial hamstring SLR was significantly increased after WBV (p = 0.039 and p = 0.043, respectively). No significant differences were found for the hamstring MLR size after WBV. Maximum tibial translation was significantly decreased after WBV (p = 0.031). Our results suggest that single WBV exposure has a positive effect on knee joint stability as a short-term adaptation on neuromuscular level. This appears to be directly associated with an increase of hamstring SLR size in response to the anterior tibial movement which may cause the decrease in anterior tibial translation. PMID- 18401810 TI - Exercise training is beneficial for Alzheimer's patients. AB - Decreased ability to perform activities of daily living (ADLs) associated with deterioration in physical capacity are key determinants of the poor quality of life and loss of independence of patients with Alzheimer's disease (AD). The purpose of this study was to determine the effects of a 12-week training program (including resistance, flexibility, joint mobility and balance/coordination exercises) for Spanish patients with AD on their i) overall functional capacity (muscle strength and flexibility, agility and balance while moving, and endurance fitness), and ii) ability to perform ADLs. Using a randomized block design, 16 patients were assigned to a training (mean [SD] age: 76 [4] yrs) or control group (73 [4] yrs) (n = 8 subjects [3 male, 5 female] per group). The results showed significant improvements after training (p < 0.05) in upper and lower body muscle strength and flexibility, agility and dynamic balance, and endurance fitness (using the Senior Fitness test), gait and balance abilities (with subsequent decrease in risk of falls) (Tinetti scale) and in the ability to perform ADLs independently (Katz and Barthel scores). No changes (p > 0.05) were found in the control group over the 12-week period. Exercise training could be included in the overall medical/nursing care protocol for patients with AD. PMID- 18401811 TI - Athletes' knowledge and views on OTC medication. AB - A questionnaire was administered to elite athletes from Australia, Canada, the UK, and the USA representing 10 Olympic sports in order to explore knowledge and understanding of over-the-counter (OTC) medication since the removal of many of these substances from the World Anti-Doping Agency (WADA) Prohibited List, in 2004. Athletes demonstrated limited knowledge and understanding. Around half (50.5 %) knew the penalty incurred following a doping violation involving a banned OTC stimulant. The terms Monitoring Program and Specified Substance List were understood by 43.3 % and 67.5 % of respondents, respectively. Overall, the status of substances in relation to the Prohibited List was correctly identified in just 35.1 % of cases. As a whole, athletes were of the opinion that OTC stimulants posed a risk to health, were performance enhancing and that their use was against the spirit of sport. They were undecided as to whether these drugs should be returned to the Prohibited List. Elite athletes require targeted education programmes that will enable them to make informed decisions on the potential of OTC medications for therapeutic or performance enhancing purposes. PMID- 18401812 TI - [Nasal dermoid sinus cyst: accidental coincidence or syndrome association?]. AB - BACKGROUND: Peters anomaly is a rare congenital glaucoma disease. The Peters' plus syndrome is characterized by distinct malformations. As some of the common craniofacial malformations like cleft lip and palate are frequent in Peters' plus syndrome, no nasal dermoid sinus cysts has been reported so far. Nasal dermoid sinus cysts usually present in isolation, although associations to other anomalies or syndromes are possible. The occurrence of such an anomaly may be either accidental, or present a syndrome association. PATIENTS AND METHOD: One patient with an unilateral cleft lip and Peters' plus syndrome had undergone removal of nasal dermoid sinus cyst previously and was referred for management of recurrent disease. Complete surgical removal and plastic reconstruction was performed. RESULTS: Concerning the common (lateral) cleft lip nasal deformity with no midline nasal masses, there are reasons for the assumption that a coincidence of both anomalies might be accidental. Especially in Peters' plus syndrome no occurrence of nasal dermoids has thus far been documented. However, the embryological pathway of the frontonasal region differs from lip and palate development in time and location: So unique formation of both lesions seems inconsistent. Complete surgical removal and plastic reconstruction simultaneously or in a second step are recommended. CONCLUSION: As two cases of arhinia and Peters anomaly have been described in 1978, midline nasal masses might be a possible appearance of Peters' plus syndrome. PMID- 18401813 TI - Drug utilisation in very preterm infants: any changes during the past decade? AB - BACKGROUND: Improved standards of perinatal care for preterm infants led to decreased hospital mortality rates during the past decade. However, studies investigating changes in drug utilisation in neonatal intensive care units (NICU) during this period are missing. OBJECTIVE: The aim of the present study therefore was to evaluate the most frequently used groups of drugs in preterm infants treated in NICUs and to analyse potential changes in drug utilisation over a period of ten years. METHODS: Drug utilisation patterns in 164 preterm infants born between 1989 and 1990 (group I; gestational age 27.2+/-1.2 weeks, birth weight 970+/-145 g) were compared to those in 113 preterm infants born between 2001 and 2004 (group II; gestational age 26.9+/-1.65 weeks, birth weight 930+/ 253 g, mean and standard deviation each) with need for postnatal mechanical ventilation. RESULTS: Significant changes in drug utilisation patterns were observed for complete courses of antenatal corticosteroids (40 vs. 51.5%), diuretics (78 vs. 36.6%), surfactant (63.3 vs. 75%), methylxanthines (89.9 vs. 56.7%), sedatives/analgesics (82.4 vs. 91.5%) and catecholamines (38.3 vs. 52.4%) (group II vs. group I each). Postnatal corticosteroids were applied more often in group II (17.4 vs. 13.4%). However, duration of postnatal corticosteroid treatment has decreased (6 d vs. 13 d). The use of antibiotics remained unchanged (100 vs. 98.9%). Comparison of clinical outcome variables showed a decreased duration of mechanical ventilation and a significantly increased survival rate. CONCLUSION: Drug utilisation patterns in preterms have changed considerably during the past decade. Improved standards of care and shorter duration of mechanical ventilation may be operative. PMID- 18401814 TI - Testicular histology related to fertility outcome and postpubertal hormone status in cryptorchidism. AB - BACKGROUND: Early surgical correction of an undescended testis is performed to prevent the development of male infertility. However, in boys with cryptorchidism early successful surgery cannot prevent infertility if they lack Ad spermatogonia. In this study, sperm concentrations and postpubertal hormone levels were correlated to bilateral testicular histology. The aim was to define the risk of future infertility via a testis biopsy program for boys with cryptorchidism. METHODS: Eighty-nine boys who had an orchidopexy were subjected to bilateral testicular biopsy. Histological analysis of 178 biopsies indicated three groups of high, intermediate, and low risk of infertility according to the presence of Ad spermatogonia. After puberty, sperm concentrations were analysed and correlated with plasma gonadotropin and testosterone levels. FINDINGS: In patients with unilateral cryptorchidism 70% of scrotal testes had an impaired transformation of Ad spermatogonia, indicating that cryptorchidism is a bilateral disease. Sperm concentrations correlated to the number of Ad spermatogonia found at the time of orchidopexy (p<0.001). All males in the high risk of infertility group were oligospermic (mean: 8.9 x 10 (6) sperm/ejaculate) and 20% were azoospermic. These patients had 25 times less sperm compared to the group with presence of Ad spermatogonia in both testes (p<0.001). Correlations between testicular histology and postpubertal hormone levels confirmed a relative gonadotropin deficiency in the majority of males with cryptorchidism. INTERPRETATIONS: Ad spermatogonia proved to be a discriminating factor for the fertility outcome in cryptorchidism. Gonadotropin treatment following orchidopexy should be considered in cryptorchidism when no Ad spermatogonia are found in undescended gonads and scrotal testis have Ad germ cell counts <0.005 per tubule. PMID- 18401815 TI - Intraoperative diffusion-weighted imaging for visualization of the pyramidal tracts. Part I: pre-clinical validation of the scanning protocol. AB - Integration of intraoperative diffusion-weighted imaging (iDWI) into neuronavigation can be potentially useful for identification of the pyramidal tract during surgery for parenchymal brain lesions. The technique of iDWI using an intraoperative MR scanner of low magnetic field strength (0.3 Tesla) has been developed. For image acquisition, a specially designed solenoid radiofrequency receiver coil integrated with a modified Sugita head holder (head-holder coil) was used. While the sensitivity characteristics of the head-holder coil were found to be 29% lower compared to a diagnostic quadrature head coil, they were sufficient to obtain iDWI images of good quality. The relationship between the angle of the motion probe gradient (MPG) pulse to the vertical axis and pyramidal tract contrasting were examined in 4 healthy men with a mean age of 30+/-5.7 years. The contrast ratio reached a maximum when the MPG pulse was applied exactly in the anteroposterior direction. The difference of the contrast ratio between right and left sides was not statistically significant. Pyramidal tract visualization became worse and the contrast ratio was reduced when the MPG pulse was applied at different angles to the vertical axis; the reduction rate varied from 20.1 to 27.9% for each 15 degrees of rotation irrespective of its side. In conclusion, the developed scanning protocol for iDWI using an originally designed head-holder coil allowed effective visualization of the pyramidal tracts using an intraoperative MR scanner of low magnetic field strength. PMID- 18401816 TI - Intraoperative diffusion-weighted imaging for visualization of the pyramidal tracts. Part II: clinical study of usefulness and efficacy. AB - Precise identification and preservation of the pyramidal tract during surgery for parenchymal brain tumors is of crucial importance for the avoidance of postoperative deterioration of the motor function. The technique of intraoperative diffusion-weighted imaging (iDWI) using an intraoperative MR scanner of low magnetic field strength (0.3 Tesla) has been developed. Its clinical usefulness and efficacy were evaluated in 10 surgically treated patients with gliomas (5 men and 5 women, mean age: 41.2+/-13.9 years). iDWI permitted visualization of the pyramidal tract on the non-affected side in all 10 cases, and on the affected side in 8 cases. Motion artifacts were observed in four patients, but were not an obstacle to identification of the pyramidal tract. Good correspondence of the anatomical landmarks localization on iDWI and T (1) weighted imaging was found. All participating neurosurgeons agreed that, in the majority of cases, iDWI was very useful for localization of the pyramidal tract and for clarification of its spatial relationships with the tumor. In conclusion, image quality and accuracy of the iDWI obtained with an MR scanner of low magnetic field strength (0.3 Tesla) are sufficient for possible incorporation into an intraoperative neuronavigation system. The use of iDWI in addition to structural iMRl and subcortical functional mapping with electrical stimulation can potentially result in a reduction of the postoperative morbidity after aggressive surgical removal of lesions located in the vicinity to the motor white matter tracts. PMID- 18401817 TI - Endoscopic optic nerve decompression for idiopathic intracranial hypertension in two cases: case report. AB - Pseudotumor cerebri is a syndrome with headaches and visual loss that sometimes cannot be treated with medication. In spite of medical treatment, the clinical course in some patients can approach extreme dysfunction with severe impairment in visual field and acuity as well as incapacitating headache. Patients with progressive or persistent symptoms require surgical treatment and decompression of the optic nerve is the preferred surgical procedure. We report on two cases of bilateral decompression of the optic nerves performed through an endonasal endoscopic approach. PMID- 18401818 TI - Microsurgically induced aneurysm models in rats, part I: techniques and histological examination. AB - INTRODUCTION: The percentage of aneurysms treated surgically has steadily decreased since the results of the ISAT study were published in 2002. The aim of this study was to develop different reliable and reproducible aneurysm models for microsurgical training and further research to guarantee effective instruction in microsurgery for young neurosurgeons with comparable aneurysms like in humans. METHODS: Arterial and venous pouch aneurysm models were created microsurgically using 22 Wistar rats. The femoral and the proximal iliac vessels and the bifurcation of the common carotid artery were exposed for induction. For histological examination every aneurysm was dissected out and analysed. RESULTS: A total of 39 microaneurysms was created in three different regions. During the creation four complications occurred: 4 bleedings and 1 defect were observed and immediately treated. Linear regression curves of the microsurgical evaluation showed a significant advancement in the course of the study. The volumes of the different models were: 2.58+/-1.01 mm (3) for the carotid, 6.49+/-3.36 mm (3) for the iliac and 10.41+/-4.13 mm (3) for the femoral aneurysms. The aspect ratios were 1.86+/-0.45 at the iliac, 1.62+/-0.3 at the femoral and 1.21+/-0.29 for the carotid aneurysms. In 89.7% of the cases the aneurysm sac was thrombosed accentuated at the aneurysm tip while the central zone of blood inflow revealed no thrombus formation. The proportion of endothelial cells displayed a reduction in relation to the total number per cross-section in all aneurysm types. CONCLUSIONS: The presented aneurysm models in rats are reliably and immediately available for further training or scientific histological investigations. Despite the fact that these are not bifurcation aneurysms, basic techniques such as suturing and microtechniques used for the dissection and repair of vessels can be taught. PMID- 18401819 TI - Intra-parenchymal tumor biopsy using neuroendoscopy with navigation. AB - BACKGROUND AND OBJECTIVES: Neuroendoscopy has allowed us to biopsy tumors located in the ventricle and the para-ventricle. With this technique we can obtain specimens under direct visual monitoring and examine tumor dissemination. For intra-parenchymal tumors, however, we normally use stereotactic procedures to collect tissues or perform open biopsies. We now report that we have successfully combined neuroendoscopy with navigational guidance to biopsy intra-parenchymal tumors. We explain our methods and discuss the advantages and disadvantages of this technique. CASES AND METHODS: We carried out intra-parenchymal tumor biopsies or resection using neuroendoscopy and guided navigation in three patients. We advanced a transparent sheath, containing a removable inner tube on which a navigation tool was mounted, to target tissues. We directly monitored procedures with a neuroendoscope placed in the transparent sheath. We collected specimens just in front of and in the tumor by forceps. In one patient with bleeding, we used the aspirator tip as a monopolar coagulator. We were also able to check for bleeding along the sheath tract using endoscopy. Without causing any new neurological deficits, we collected tissues for histological diagnosis of astrocytoma in two patients and radiation necrosis in one. CONCLUSIONS: We believe that combining neuroendoscopy with navigation guidance is a safe and precise method for obtaining biopsies of intra-parenchymal tumors. Tumors with rich vasculature will not benefit from this procedure until better hemocoagulation instruments have been developed. PMID- 18401820 TI - Locomotor activity is a predictive test after global ischemia-reperfusion in Mongolian gerbils. AB - OBJECTIVE: The Mongolian gerbil is one of the main animal species used for the study of global ischemia, due to its specific Circle of Willis. Because of their anatomic variations, a large number of animals is needed. On account of the specific vulnerability of the hippocampus, striatum and neocortex, it is possible to evaluate the severity of the ischemic damage through an analysis of locomotor activity. The tests support the sensitvity of the experimental sample and compensate the interanimal variability. METHODS: The locomotor pattern of 30 male Mongolian gerbils was recorded before they were subjected to experimental bilateral carotid clippage for 15 minutes followed by reperfusion. A transparent 75x50x90 cm acrylic box was filmed in order to determine the total distance covered by the animals in five minutes, for three consecutive days. The locomotor activity of the animals was also examined in an open field at 24 hours and seven days after ischemia. Serum neurospecific enolase (NSE) was measured in the ischemic group and compared with that of an intact control group. RESULTS: The recording for normal animals was uniformly similar (average 200 squares in periphery), in the first trial of 3 consecutive days (188+/-6.7 S.D.). After ischemia, the numbers increased to 388 (+/- 40 S.D.), indicating that they were sensitive to the ischemic episode. Seven days later they returned to basal values. Serum NSE was high in the ischemic group versus the intact control group (S=<0.001). CONCLUSIONS: Locomotor activity in an open field is a useful reference as a predictive test to determine the sensitivity of experimental animals to ischemia. It is also associated to the degree of cerebral damage in global ischemia-reperfusion, and this behavior is representative of the expression of selective ischemic injury. The determination of NSE is useful as an associated parameter of ischemic injury. PMID- 18401821 TI - Evaluation of carotid plaque composition by computed tomographic angiography and black blood magnetic resonance images. AB - The purpose of this study was to evaluate quantitatively the composition of carotid plaque by computed tomographic (CT) angiography and qualitatively by black blood magnetic resonance images (MRI). Thirty-eight patients with high grade carotid artery stenosis were included in this study. Ultrasonography, CT angiography and black blood MRI of the cervical carotid artery were performed, and the CT number was measured in Hounsfield units (HU). The average CT number of the 15 unstable plaques (39.5%) was 27.7+/-7.5 HU and that of the 23 stable plaques (60.5%) was 60.4+/-20.8 HU (p=0.0001). In the 23 patients with stable plaque, 21 demonstrated isointensity in T (1)- and T (2)-weighted images in the black blood MRI (p=0.0001). By using CT angiography and MRI, precise images of the pathology of the carotid arterial wall can be obtained. It is possible to evaluate the components of carotid artery plaque with high reliability by quantification of the CT number in CT angiography and performing black blood MRI as well as carotid ultrasonography. PMID- 18401822 TI - Combined subtemporal and retrosigmoid keyhole approach for extensive petroclival meningiomas surgery: report of experience with 7 cases. AB - OBJECTIVE: The resection of petroclival meningiomas offers great challenges to the neurosurgeon. Surgery via a combined subtemporal and retrosigmoid keyhole approach surgery was evaluated for the treatment of extensive petroclival meningiomas on the basis of our experience with 7 cases. METHODS: From July 2002 to July 2005, the resections of 7 petroclival meningiomas, which involved both supra- and infra-tentorial regions, were performed via a combined subtemporal and retrosigmoid keyhole approach. The extent of tumor resection was evaluated by MRI 3 months after surgery, and postoperative complications were investigated. RESULTS: The maximum diameter of tumors ranged from 3.4 to 6.0 cm (mean: 4.4 cm). Gross total resection (GTR) was achieved in 3 cases, giving a GTR rate of 43%. Subtotal resection (STR) was carried out in 4 cases. Neurological status remained intact in one case, while the others presented with cranial nerve deficits (VII, VI, V, III and lower CN). No death was reported during the postoperative period. CONCLUSION: This combined keyhole approach is suitable for the treatment of extensive petroclival meningiomas. It provides easy and quick access to the supra and infratentorial juxta-clival regions without any petrous bone drilling. Complications related to the approach can be minimized. PMID- 18401823 TI - Comparison of techniques for decompressive lumbar laminectomy: the minimally invasive versus the "classic" open approach. AB - OBJECT: Decompressive laminectomy offers an effective surgical treatment of lumbar spinal stenosis. The purpose of this study was to compare the elements of treatment commonly associated with successful outcomes in the assessment of laminectomies - operating room times, estimated blood loss, length of stay, and complications - of the minimally invasive and open approach laminectomies. METHODS: We retrospectively reviewed the medical records and relevant imaging of 126 patients who underwent surgical decompression for lumbar stenosis. Thirty eight patients underwent bilateral decompression via a unilateral minimally invasive technique, while 88 patients underwent bilateral decompression via a standard open technique. A chart review was performed to determine intraoperative blood loss, length of operative time, length of hospital stay, and number and nature of complications. RESULTS: The minimally invasive lumbar laminectomy (MID) patients had shorter operating room times, less estimated blood loss, shorter length of stay, and fewer complications. CONCLUSIONS: Bilateral decompression of lumbar spinal stenosis via a unilateral approach involves shorter operating times and less blood loss, less muscle dissection, fewer and less severe complications, and better mobility in the immediate postoperative period than open decompressive techniques. In addition, this technique is very similar to the commonly performed microendoscopic discectomy and is easily mastered over time. PMID- 18401824 TI - Percutaneous anterior odontoid screw fixation. AB - OBJECTIVE: A technical description of a novel percutaneous technique of anterior odontoid screw fixation is given and the clinical and radiological results of this technique in 5 patients are described. METHOD: The percutaneous anterior odontoid screw fixation technique was described as a cadaveric study in 1999. To the best of our knowledge, no patient series operated on by this technique has so far been presented in the English literature. We have percutaneously operated on 5 patients with unstable odontoid fractures between February 2004 and July 2006. RESULTS: There have not been any complications in our patients. The first four patients showed radiological evidence of fusion in their latest control. CONCLUSIONS: Percutaneous anterior odontoid screw fixation is a minimally invasive and feasible surgical procedure. PMID- 18401825 TI - Is it appropriate to redefine the indication for stereotactic brain biopsy in the MRI Era? Correlation with final histological diagnosis in supratentorial gliomas. AB - BACKGROUND: The aim of this study was to evaluate the advisability of modifying the indications for stereotactic brain biopsy (SBB) in high- and low-grade supratentorial glial tumors in correlation with the diagnostic accuracy of magnetic resonance imaging (MRI). MATERIALS AND METHODS: We analyzed 174 cases of supratentorial glial lesions diagnosed by MRI before biopsy. On the basis of the MRI findings the patients were divided into two groups: high-grade (n=107) and low-grade (n=67) gliomas. All patients underwent CT-guided biopsy. Only one preoperative diagnosis was allowed. RESULTS: A final histological diagnosis was achieved in 95% of the 174 cases. In the group of high-grade gliomas (HGG) there was diagnostic coincidence in 87% of cases, reaching 100% in lesions of the corpus callosum. In 11 cases (10%) the histological analysis changed the presumptive diagnosis and the consequent management. In the group of low-grade gliomas (LGG) there was diagnostic coincidence in 63% (42 cases), whereas there was discordance in 30%: 10 cases were upgraded to anaplastic astrocytoma, and in 10 cases no tumors were observed at all. CONCLUSIONS: In the future, the histological diagnosis of glial tumors will include molecular genetic definition, thus making it crucial for management using the new therapeutic options. Today, the indications for biopsy in lesions mimicking high-grade gliomas are mainly linked to the site of the tumor, coexisting differential diagnoses or more than one treatment option. In lesions of the corpus callosum, histological analysis seems not to be required in order to proceed to complementary therapies. On the contrary, in lesions where MRI findings indicate low-grade gliomas, grading is crucial also in order to avoid treatment inappropriate in non-neoplastic lesions. PMID- 18401826 TI - Frame-based stereotactic resection of subcentimeter arteriovenous malformations in deep or eloquent regions of the brain: indications and technique with eight consecutive operations. AB - Subcentimeter arteriovenous malformations (AVMs) located in deep or eloquent cortex can be difficult to localize intraoperatively and safely remove with surgery. Nevertheless, surgical resection may be the optimal definitive treatment option available for select patients. In this communication, we describe our experience using a framed-based stereotactic approach for resecting these AVMs. The operative records of all AVMs treated at our institution over an 8-year period (1996-2004) were reviewed. 180 surgically treated AVMs were identified. From this group of patients, frame-based stereotaxy was used for 8 AVMs (4.4%) in 7 patients. The angiograms, operative reports, and medical records for these 7 patients were retrospectively reviewed with attention to neurological outcome, extent of AVM obliteration, and anatomic factors that impacted the decision to employ a frame-based stereotactic approach. All AVMs removed with this technique were less than 1 cm in diameter. Angiography confirmed complete resection in all cases. No new neurological deficits occurred in any patient. By providing highly accurate three-dimensional nidus localization and minimizing approach-related brain manipulation, frame-based stereotaxy reduces the morbidity associated with resection of subcentimeter AVMs located in deep or eloquent regions of the brain. This technique makes a definitive surgical cure available to patients who otherwise would only be considered for radiosurgery. PMID- 18401827 TI - Course of the bony canal associated with high-positioned supraorbital foramina: an anatomic study to facilitate safe mobilization of the supraorbital nerve. AB - OBJECTIVE: In the dissection of the superior orbital rim, the supraorbital foramen must be released to preserve the supraorbital nerve. The aim of this study was to clarify the spatial dimensions of a high-positioned foramina to allow for the safe performance of this maneuver. METHODS: We examined 90 orbital sides. In the detected foramina we measured the distance between the superior orbital rim and the inferior margin of the foramen (height), and between the rim and the anterior margin of the opening of the foramen in the orbital roof (depth). To evaluate the inclination of the canal, we calculated the height:depth ratios. Foramina with a height exceeding 2.0 mm were defined as high-positioned and measurements between low- and high-positioned foramina were compared. RESULTS: We were able to find 37 foramina in 32 orbital sides; 25 (67.6%) were low-positioned and their height ranged from 0.5-1.9 mm (mean: 1.16 mm); 12 (32.4%) were high-positioned with a height ranged from 2.0-11.9 mm (mean: 4.0 mm). The depth in low- and high-positioned foramina ranged from 0.5-3.2 mm (mean: 1.44 mm) and 0.7-6.5 mm (mean: 2.13 mm), respectively. The height:depth ratio was greater in high-positioned foramina, ranging from 1.03-3.38 (mean: 2.19), than in low-positioned foramina where it ranged from 0.4-2.2 (mean: 0.98). CONCLUSION: The height:depth ratio in high-positioned foramina, an approximate mean value of 2, and the absence of a value less than 1, was considered to indicate a steep canal inclination. To avoid postoperative forehead numbness, anatomic information regarding the course of the bony canal must be considered. PMID- 18401828 TI - Modified irrigation hose placement in draping for craniotomy: provision of a free foot space for surgeons: technical note. AB - Sterility and utility are essential in surgical draping. For craniotomy, we modified the course of the irrigation hose to maintain a free foot space for the surgeon by connection with a suction bottle placed beside the patient's body through a slit made in the linen. This minor modification provides convenience to the surgeon during operations. PMID- 18401829 TI - Use of Grotenhuis perforator in endoscopic third ventriculostomy and cyst fenestration. AB - We performed a retrospective review of consecutive patients at our institution who underwent endoscopic third ventriculostomy (E3 V) or fenestration of intraventricular cysts using the Grotenhuis endoscopic perforator. The procedure was performed on 23 patients between 2001 and 2006, and included 20 E3Vs and three intraventricular cyst fenestrations. The Grotenhuis perforator was effective in accomplishing a fenestration with multiple attempts. When the floor of the third ventricle was translucent, the perforator was effective with the least amount of effort. The instrument was less effective and additional instruments were necessary in patients with arachnoid cysts or when the floor of the third ventricle was thick. The main advantage in using the Grotenhuis perforator was in displacing the floor of the third ventricle away from the basilar artery during perforation. No basilar artery injury or other serious complications occurred in patients who underwent E3 V or cyst fenestration using the Grotenhuis perforator. PMID- 18401830 TI - Protein kinase A subunit expression is altered in Bloom syndrome fibroblasts and the BLM protein is increased in adrenocortical hyperplasias: inverse findings for BLM and PRKAR1A. AB - Bloom syndrome is a genetic disorder associated with chromosomal instability and a predisposition to tumors that is caused by germline mutations of the BLM gene, a RecQ helicase. Benign adrenocortical tumors display a degree of chromosomal instability that is more significant than benign tumors of other tissues. Cortisol-producing hyperplasias, such as primary pigmented nodular adrenocortical disease (PPNAD), which has been associated with protein kinase A (PKA) abnormalities and/or PRKAR1A mutations, also show genomic instability. Another RecQ helicase, WRN, directly interacts with the PRKAR1B subunit of PKA. In this study, we have investigated the PRKAR1A expression in primary human Bloom syndrome cell lines with known BLM mutations and examined the BLM gene expression in PPNAD and other adrenal tumor tissues. PRKAR1A and other protein kinase A (PKA) subunits were expressed in Bloom syndrome cells and their level of expression differed by subunit and cell type. Overall, fibroblasts exhibited a significant decrease in protein expression of all PKA subunits except for PRKAR1A, a pattern that has been associated with neoplastic transformation in several cell types. The BLM protein was upregulated in PPNAD and other hyperplasias, compared to samples from normal adrenals and normal cortex, as well as samples from cortisol- and aldosterone-producing adenomas (in which BLM was largely absent). These data reveal an inverse relationship between BLM and PRKAR1A: BLM deficiency is associated with a relative excess of PRKAR1A in fibroblasts compared to other PKA subunits; and PRKAR1A deficiency is associated with increased BLM protein in adrenal hyperplasias. PMID- 18401831 TI - Epidermal growth factor and parathyroid hormone-related peptide mRNA in the mammary gland and their concentrations in milk: effects of postpartum hypoxia in lactating rats. AB - The physiological adaptations of the neonatal rat to hypoxia from birth include changes in gastrointestinal function and intermediary metabolism. We hypothesized that the hypoxic lactating dam would exhibit alterations in mammary gland function leading to changes in the concentration of milk peptides that are important in neonatal gastrointestinal development. The present study assessed the effects of chronic hypoxia on peptides produced by the mammary glands and present in milk. Chronic hypoxia decreased the concentration of epidermal growth factor (EGF) in expressed milk and pup stomach contents and decreased maternal mammary gland EGF mRNA. The concentration of parathyroid hormone-related protein (PTHrp) was unchanged in milk and decreased in pup stomach contents; however, mammary PTHLH mRNA was increased by hypoxia. There was a significant increase in adiponectin concentrations in milk from hypoxic dams. Chronic hypoxia decreased maternal body weight, and pair feeding normoxic dams an amount of food equivalent to hypoxic dam food intake decreased body weight to an equivalent degree. Decreased food intake did not affect the expression of EGF, PTHLH, or LEP mRNA in mammary tissue. The results indicated that chronic hypoxia modulated mammary function independently of hypoxia-induced decreases in maternal food intake. Decreased EGF and increased adiponectin concentrations in milk from hypoxic dams likely affect the development of neonatal intestinal function. PMID- 18401832 TI - Sustained efficacy and reduced hypoglycemia during one year of treatment with vildagliptin added to insulin in patients with type 2 diabetes mellitus. PMID- 18401833 TI - The double role of epicardial adipose tissue as pro- and anti-inflammatory organ. AB - Obesity is associated with low grade inflammation. Whether this is just an adaptive response to excess adiposity to maintain a normal oxygen supply or a chronic activation of the innate immune system is still unknown. Recent research has focused on the origin of the inflammatory markers in obesity and the extent to which adipose tissue has a direct effect. The production of adipokines by visceral adipose tissue is of particular interest since their local secretion by visceral fat depots may provide a novel mechanistic link between obesity and the associated vascular complications. Growing evidences suggest that the epicardial adipose tissue, the visceral fat depot located around the heart, may locally interact with myocardium and coronary arteries. Epicardial fat is a source of adiponectin and adrenomedullin, adipokines with anti-inflammatory properties, and several proinflammatory cytokines as well as Tumor Necrosis Factor-alpha (TNF alpha), Interleukin 1 (IL1), IL-1 h, Interleukin (IL6), Monocyte Chemoattractive Protein-1 (MCP-1), Nerve Growth Factor (NGF), resistin, Plasminogen Activator Inhibitor-1 (PAI-1), and free fatty acids. Epicardial adipose tissue could locally modulate the heart and vasculature, through paracrine secretion of pro- and anti-inflammatory cytokines, thereby playing a possible role in the adiposity related inflammation and atherosclerosis. On the other hand, epicardial fat could exert a protective effect through adiponectin and adrenomedullin secretion as response to local or systemic metabolic or mechanical insults. Future studies will continue to provide new and fascinating insights into the double role of epicardial adipose tissue in the development of cardiovascular pathology and/or in protecting the heart and arteries. PMID- 18401834 TI - Maternal perinatal undernutrition impairs chromaffin cells proliferation in the postnatal rat. AB - Numerous data show that malnutrition during early life programs chronic diseases in adulthood. Many of these disorders may result from alterations in the development of neuroendocrine systems, such as the hypothalamo-pituitary-adrenal axis and the sympathoadrenal system. We have previously reported that maternal 50% food restriction during late pregnancy and lactation reduces adrenal weight and impairs chromaffin cell differentiation in male rats at weaning. In addition, maternal undernutrition modifies the expression of several genes involved in proliferation and apoptosis. This study therefore investigated the impact of maternal food restriction on adrenal cell growth in the late postnatal rat. Histological analysis showed that the number of proliferating chromaffin cells assessed by nuclear labelling with BrdU was reduced by 45%, whereas the level of apoptosis visualised by caspase-3 immunoreactivity was increased by 340% in adrenal medulla of offspring from undernourished mothers. In contrast, maternal food restriction did not affect proliferation and apoptosis in cortical cells of rats. These developmental changes were associated with overexpression of TGFbeta2. These data show that perinatal undernutrition impairs the balance between chromaffin cell proliferation and apoptosis. These modifications may lead to "malprogramming" of adrenal medulla development, which could contribute to the pathogenesis of chronic diseases in adulthood. PMID- 18401835 TI - White adipose tissue production and release of IL-6 and TNF-alpha do not parallel circulating and cerebrospinal fluid concentrations in pregnant rats. AB - IL-6 and TNF-alpha are synthesized in white adipose tissue both by adipocytes and by the stroma-vascular fraction. They both are known to interfere with insulin signaling, reducing insulin sensitivity and lipid deposition. At a central level, IL-6 enhances sympathetic nervous system activity, thus enhancing lipolysis and reducing fat mass. During late pregnancy, white adipose tissue (WAT) mass increases and insulin sensitivity decreases. To assess the involvement of both adipokines in such processes, we analyzed the tissue content and release of both adipokines in parametrial and subcutaneous WAT depots and their circulating and cerebrospinal fluid concentrations in nonpregnant rats and in pregnant rats by days 8, 15, and 19 of pregnancy. The tissue content of both adipokines was enhanced 5-6 times by day 8 until the end of pregnancy in parametrial WAT, whereas the increase took place by day 15-19 in subcutaneous WAT. No increase in tissue release was detected, suggesting a local action. However, circulating IL-6 concentration was enhanced by day 8 until the end of pregnancy, suggesting sources other than WAT. IL-6 concentration in cerebrospinal fluid paralleled the increases in serum by days 8 and 15, suggesting a systemic origin. However, it returned to basal levels by day 19, suggesting a central control for IL-6 entrance. TNF-alpha was not detected in either serum or cerebrospinal fluid. These results led us to conclude that across pregnancy adipokines control WAT depots in a time- and depot-dependent manner. They do so directly, by local production, but the enhanced concentrations of both circulating and CSF IL-6 suggest an indirect action mediated by the nervous system. PMID- 18401836 TI - Effect of moderate-intensity exercise session on preprandial and postprandial responses of circulating ghrelin and appetite. AB - Responses of plasma total ghrelin and appetite were investigated during preprandial and postprandial stages of recovery from a moderate-intensity cycling session. Healthy recreationally active men underwent one exercise and one control trial. In the exercise trial, subjects exercised for approximately 60 minutes, while in the control trial they rested quietly for the same duration. After the intervention, subjects rested for 120 minutes and then consumed a test meal. Measurements were obtained immediately and 120 minutes after the intervention and then during 180 minutes of the postprandial period. The post-intervention concentration of total ghrelin was lower (p<0.05) in the exercise than in the control trial. The modulating effect of exercise was related to the reduction in the postprandial rather than preprandial concentration. Post-intervention scores of appetite were not different between the two trials, but when preprandial and postprandial responses were considered separately, postprandial hunger and desire to eat was higher (p<0.05) in the exercise trial. In summary, during recovery from moderate-intensity exercise, total ghrelin does not respond in a compensatory manner to disturbances in energy balance. Thus, an exercise-induced increase in appetite during the later stages of recovery coinciding with the postprandial state cannot be explained by changes in the plasma concentration of total ghrelin. PMID- 18401837 TI - Type 1 diabetes alters brain cannabinoid receptor expression and phosphorylation status in rats. AB - One of the most common symptoms of diabetes is extreme hunger, but the brain mechanism underlying this hyperphagia is unknown. The endocannabinoid system has emerged as one of the main food intake regulators in the brain. However, the effects of type 1 diabetes on the endocannabinoid system are not completely known. Thus, the aim of the present work is to establish the possible alterations induced by type 1 diabetes on the brain endocannabinoid system in rats. Western blot and immunocytochemistry were used to measure CB1 and phosphorylated CB1 receptor expression in several prosencephalic regions in streptozotocin-induced type 1 diabetic rats. Serum leptin levels were measured by ELISA. CB1 receptor expression was increased in striatum and hypothalamus of diabetic animals, with no changes in other brain areas studied. CB1 receptor phosphorylation was also increased in the same brain areas. Type 1 diabetes induced significant weight loss, and serum leptin levels were severely decreased. These results reinforce the possible role of the CB1 receptor as a pharmacological target for the clinical management of appetite in diabetic patients. PMID- 18401838 TI - Prevalence of risk determinants for metformin-associated lactic acidosis and metformin utilization in the study of health in pomerania. AB - Risk determinants for the life threatening complication of metformin-associated lactic acidosis are frequently disregarded. Our first aim was to investigate the prevalence of risk determinants in subjects with type 2 diabetes mellitus (DM2) taking metformin compared to subjects with nonmetformin treatment. Our second aim was to estimate the proportion of subjects with alternative drug-treatment, and no risk determinants, which would probably benefit from metformin. The Study of Health in Pomerania is a population-based health survey including 322 DM2 subjects. Risk determinants were assessed by personal interview, ultrasound, and laboratory analysis. Among the subjects with DM2 n=92 (28.6%) were treated with metformin, n=162 (50.3%) with alternative medication, and n=68 (21.1%) with diet. The prevalence of at least one risk determinant was 65% [corrected] for metformin users. There was no difference in number and type of risk determinants. Heart failure, angina pectoris, and liver disorders presented the most frequent risk determinants. Current risk determinants for metformin-associated lactic acidosis are largely disregarded. Improved selection of patients can result in safe metformin utilization in one quarter of subjects on DM2 related drug treatment. Risk determinants need to be revised. A more practical definition of risk determinants would improve prescription adherence. PMID- 18401839 TI - Retinol-binding protein 4 and nicotinamide phosphoribosyltransferase/visfatin in rat obesity models. AB - Retinol-binding protein 4 (RBP4) and nicotinamide phosphoribosyltransferase/visfatin (Nampt/visfatin) are adipocyte-secreted proteins (adipokines) whose relevance to the metabolic syndrome and regulation in obesity remain controversial. Here, we tested the hypothesis that adipose tissue expression and circulating levels of these two adipokines are elevated in obesity by analyzing their changes in both a genetic and a diet-induced model of obesity in the rat (obese FA/ FA Zucker rats and Wistar rats fed a cafeteria diet, respectively). Compared with lean controls, obese FA/ FA rats were hyperleptinemic, hyperinsulinemic, and insulin resistant and had reduced RBP4 serum levels and mRNA levels in adipose depots, unchanged Nampt/visfatin serum levels, and reduced Nampt/visfatin mRNA levels selectively in the inguinal adipose depot. Cafeteria diet-induced obesity resulted in increased fed blood glucose levels, a variable degree of insulin resistance, unchanged serum Nampt/visfatin and RBP4 levels, and reduced mRNA levels of both adipokines in several adipose depots. Hence, increases in RBP4 or Nampt/visfatin do not accompany obesity and insulin resistance in the models examined. PMID- 18401840 TI - The use of 17 alpha-hydroxyprogesterone caproate (17p) in women with cervical cerclage. AB - Our objective was to compare the incidence of recurrent spontaneous preterm delivery (SPTD) in patients with cervical cerclage treated with weekly 17 alpha hydroxyprogesterone caproate (17P) injections versus daily outpatient nursing surveillance (ONS) without 17P. Included in this retrospective cohort study were singleton gestations with cerclage placed at the discretion of the provider due to prior SPTD, delivering between January 1, 2004 and May 1, 2006. The study group (n = 232) consisted of women receiving once-weekly nursing visit and 17P injection. The control group (n = 1650) consisted of women enrolled for ONS (twice-daily electronic uterine contraction monitoring and nursing assessment). Data were further stratified by the number of prior preterm deliveries (1, > 1). Primary study outcome was the incidence of SPTD. No difference in rates of recurrent SPTD at < 37 or < 35 weeks were observed between the study and control groups. Study patients were less likely to be diagnosed with preterm labor (PTL) than controls (45.7% versus 70.8%, respectively; P < 0.001). The incidence of preterm premature rupture of membranes was similar between the groups (8.6% versus 8.1%; P = 0.770). We concluded that the incidence of recurrent SPTD was similar in women with cerclage treated with 17P or ONS, although women receiving 17P had a lower incidence of PTL. This benefit of 17P should be considered when managing patients with prior SPTD and cerclage. PMID- 18401841 TI - Fetal cardiac malposition: incidence and outcome of associated cardiac and extracardiac malformations. AB - Cardiac malposition is a rare but important finding when detected on fetal ultrasound. The purpose of this study was to evaluate the incidence of fetal cardiac malposition, associated abnormalities, and clinical outcome in a tertiary care medical center. Records of fetuses (1993 to 2006) with dextroposition, dextrocardia, mesocardia, ectopia cordis, or heterotaxy were reviewed. The presence of congenital heart disease (CHD), extracardiac anomalies, and outcome were noted. Cardiac malposition was present in 101 fetuses among a total of 3313 (3%) pregnancies. In 78 (78%) patients, the heart was positioned in the right hemithorax. Of those, 26 (33%) had dextrocardia (CHD = 21), and 52 (67%) had dextroposition (CHD = 14). Sixteen (16%) patients had mesocardia (CHD = 8), and 7 (7%) had ectopia cordis (CHD = 6). The majority (58%) of fetal cardiac malposition was caused by intrathoracic masses. Concomitant CHD occurred in 50%. Outcome was available in 97%. The perinatal and neonatal mortality rate was 30%; the elective termination rate was 4%. Patients with CHD had higher mortality rates. The highest mortality rates occurred in ectopia cordis and combined disease of diaphragmatic hernia and CHD. PMID- 18401842 TI - Isolation and characterization of a novel trypsin inhibitor from fresh lily bulbs. AB - A 17-kDa trypsin inhibitor was isolated from fresh lily bulbs with an isolation procedure that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Sepharose, and gel filtration by FPLC on Superdex 75. Its N-terminal sequence displayed similarity to a short segment of the sequences of the Populus tremula trypsin inhibitor, a putative trypsin inhibitor from Arabidopsis thaliana and sporamin B from sweet potato. The trypsin inhibitor was adsorbed on DEAE-cellulose, unadsorbed on Affi-gel blue gel, and adsorbed on SP-Sepharose. It dose dependently inhibited trypsin with an IC (50) value of 1.3 microM. There was a stimulatory effect on macrophage production of nitric oxide. Unlike field bean trypsin inhibitor it did not inhibit [methyl-(3)H]thymidine incorporation by leukemia L1210 cells and MBL2 cells when tested up to 100 microM. In contrast to broad bean trypsin inhibitor, there was no inhibitory effect on HIV-1 reverse transcriptase when lily bulb trypsin inhibitor was tested up to 100 microM. The present report is one of the very few on bulbs in contrast to the voluminous literature on seeds. PMID- 18401843 TI - Induction of mitochondrial-dependent apoptosis by an essential oil from Tanacetum gracile. AB - The essential oil of Tanacetum gracile (Accession no. AT-01 termed AT-01 in the manuscript), a cold desert alpine highly aromatic herb, has 40 constituents including lavendulol (21.5 %), lavendulol acetate (1.7 %), alpha-pinene (11.2 %), 1,8-cineole (15.2 %), CIS-beta-ocimene (6.9 %), borneol (6.1 %), limonene (5.1 %) and chamazulene (3.7 %). AT-01 was evaluated for its anticancer activity. It inhibited HL-60 cell proliferation with an IC (50) of 27 microg/mL. Furthermore, AT-01 induced apoptosis in human leukemia HL-60 cells as measured by several biological end points. AT-01 induced apoptotic body formation, enhanced annexinV FITC binding of the cells, increased sub-G (0) DNA fraction, loss of mitochondrial membrane potential (Deltapsi (mt)) and release of cytochrome c from mitochondria, activated caspase-9 as well as caspase-3, and increased cleavage of PARP in HL-60 cells. Thus, AT-01 induced apoptosis through the mitochondrial dependent pathway in HL-60 cells. PMID- 18401844 TI - LC-DAD-MS (ESI+) analysis and antioxidant capacity of crocus sativus petal extracts. AB - In this study, various fractions isolated from the petals of Crocus sativus were assessed at first for their phenolic content both qualitatively and quantitatively and secondly for their antioxidant activity. The phytochemical analysis was carried out by LC-DAD-MS (ESI (+)) whereas the antioxidant potential was evaluated by applying two methodologies, the DPPH. radical scavenging activity test and the Co(II)-induced luminol chemiluminescence procedure. According to data obtained from these antioxidant tests, the diethyl ether, ethyl acetate and aqueous fractions demonstrated the strongest antioxidant capacity. Interestingly, the major constituents identified in these fractions correspond to kaempferol, quercetin, naringenin and some flavanone and flavanol derivatives glycosylated and esterified with phenylpropanoic acids. In addition, the presence of some nitrogen-containing substances, as well as other phenolics and phenylpropanoic derivatives was also traced. The identification and structural elucidation of all substances isolated in this study was achieved by both comparing available literature data and by proposed fragmentation mechanisms based on evaluating the LC-DAD-MS (ESI (+)) experimental data. The quantitative analysis data obtained thus far have shown that Crocus sativus petals are a rich source of flavonoids. Such a fact suggests that the good antioxidant capacity detected in the various fractions of Crocus sativus petals could be attributed to the presence of flavonoids, since it is already known that these molecules exert antioxidant capability. The latter, along with the use of Crocus sativus in food and pharmaceutical industry is discussed. PMID- 18401845 TI - Two new cytotoxic pregnane glycosides from Cynanchum auriculatum. AB - Two new pregnane glycosides, kidjoranin 3- O- alpha-diginopyranosyl-(1-->4)- beta cymaropyranoside (1) and kidjoranin 3-O-beta-digitoxopyranoside (2), together with one known compound caudatin 3 -O-beta-cymaropyranoside (3), were isolated from the roots of Cynanchum auriculatum. Their structures were established on the basis of NMR analyses. Compounds 1 - 3 were tested for their in vitro inhibitory activity against the growth of human tumor cell lines SMMC-7721, HeLa and MCF7; all of them displayed marked cytotoxic activities against cells SMMC-7721 and HeLa with IC (50) values ranging from 8.6 microM to 58.5 microM, yet no activity against the cell line MCF7 was detected. PMID- 18401846 TI - New bioactive bromotyrosine-derived alkaloid from a marine sponge Aplysinella sp. AB - A new bromotyrosine-derived alkaloid, (+)-aplysinillin (1), together with seven known compounds, was isolated from the crude organic extract of the marine sponge Aplysinella sp. collected from the Federated States of Micronesia. The structure of 1 was deduced by NMR and mass spectral techniques. Compounds 1-8 were evaluated for their inhibitory activity with the hyphae formation inhibition assay in Streptomyces 85E. Compounds 1 and 8 exhibited antiproliferative activities against the human breast adenocarcinoma cancer cell line MCF-7. PMID- 18401847 TI - Dermal and transcutaneous delivery of the major glycoside constituents of Harpagophytum procumbens (Devil's Claw) in vitro. AB - The potential of the administration of Harpagophytum procumbens extract via the topical route has not been studied previously. In the current work, the dermal and transcutaneous delivery of the major pharmacologically active constituents present in H. procumbens tuber extract were determined across porcine ear skin from four vehicles: de-ionised water, 30 % ethanol in water (v/v), PEG 400, 50 : 50 PEG 400 in 30 % EtOH (v/v). Permeation profiles were obtained under infinite conditions and tape stripping was performed at 24 h. The permeation of the compounds varied according to their physicochemical properties as well as the nature of the vehicle. The highest permeation was found from the ethanol/water saturated solutions and the lowest MW harpagide was obtained at significantly higher concentrations in the receptor phase compared to the rest of the compounds, with the permeability coefficient being inversely dependent on dielectric constant of the vehicle. Depth profiling revealed higher penetration of all compounds from ethanol/water; in addition, significantly higher amounts of the pro-inflammatory harpagide were present in the strips and the remaining epidermis compared to other compounds. This suggests that ethanol is not a suitable vehicle as it leads to more harpagide penetration, potentially counteracting the anti-inflammatory activity of the other compounds. The development of new systems for local cutaneous inflammation (e. g., psoriasis, eczema) and subcutaneous inflammation (e. g., arthritis) is supported. PMID- 18401848 TI - [Management and outcome of vein of galen malformation in childhood]. AB - The vein of Galen malformation (VGM) is a rare congenital cerebral vascular malformation. In most cases it already has been diagnosed prenatally. The impact of heart failure due to the arteriovenous shunt is relevant for the prognosis.Prognosis of neonatal symptomatic VGM with early heart failure is severe. Therefore an interdisciplinary cooperation is necessary and it is important to treat patients in a clinic with a perinatal center and a neuroradiology and neurosurgery that is experienced in treating such patients. The genesis, the pathophysiology, the diagnostic agents, the therapeutic agents and the prognosis of VGM will be described. PMID- 18401849 TI - [New ways in psychiatric diagnostics?]. AB - There is an increasing criticism of the current operational diagnostic approach of ICD-10 and DSM-IV in psychiatry. It has not been possible to validate the respective diagnostic categories with neurobiological findings or to detect sharp boundaries between the diagnostic categories with respect to psychopathological symptoms. Therefore, dimensional diagnostic models are suggested in order to supplement the categorical diagnostic approach. Based on a historical review and a survey of the current criticism of the diagnostic approach of ICD-10 and DSM-IV proposals are made in order to solve the current problems. In conclusion, a multiaxial diagnostic model is suggested that comprises the axes "syndromes", "course types" and "etiology" and that may provide a bridge between the traditional categorical diagnostic approach and dimensional models. PMID- 18401850 TI - [Night eating syndrome and nocturnal eating--what is it all about?]. AB - In the first description of the night eating syndrome (NES) 1955 by Stunkard et al. the criteria included (1) consumption of at least 25% of the total calories for the day after the evening meal, (2) sleeplessness, at least until midnight more than one half of the time and (3) morning anorexia with negligible food intake at breakfast. Further studies altered these criteria step by step, without ever relating to the changes already made by other authors. So today our knowledge about NES and its related features is based on an amazing variety of constructs merely referred to by the same term. However, there seems to be an agreement about a higher prevalence of the NES in overweight and obese treatment seeking samples. The relationship between NES, body weight and a possible influence of NES on overweight and obesity remains unclear and needs to be further examined. In addition to the research activities regarding NES as a possible eating disorder, sleep disorder specialists showed a growing interest in patients with sleeplessness and nocturnal eating episodes. New definitions were developed: the "night eating/drinking syndrome" (synonymous: NES), a disorder occurring mainly in infancy and early childhood but also seen in adults. Today the less restrictive concept "sleep related eating disorder" (SRED) eliminated the NES-concept, but also states a sleep disorder that is not clearly distinguishable from NES described by several authors as a possible eating disorder. In psychosomatic research the criteria of nocturnal eating (recurrent awakenings & getting up to eat) was included in the NES by a growing number of authors in the last 15 years. Based on this diversity of diagnostic criteria in two different fields of expertise a lot of research was done do investigate the prevalence of NES and further describe patients with NES. Today a meaningful summary of these findings is not possible and despite a growing number of research in NES and obesity the clinical relevance of the concept NES remains unclear. In this article diagnostic criteria so far will be summarized and a rough differentiation of NES to related constructs and disorders will be given. PMID- 18401851 TI - [Fatigue and its determinants in radio-oncology]. AB - BACKGROUND: Fatigue is a common symptom among cancer patients, influencing their quality of life. The primary goal of his study was to identify parameters influencing fatigue, both prior to radiotherapy (RT) and changes in fatigue during RT. METHODS: 239 patients could be assessed prior to RT, 208 patients were reassessed at the end of RT. Measures comprised the Multidimensional Fatigue Inventory (MFI-20), the Resilience Scale (RS) and the SF-12 as a measure of health related Quality of Life (QoL). RESULTS: The sample revealed higher scores in the MFI and in the RS as well as lower scores in the SF-12 than normative samples. Fatigue increased significantly during RT. Fatigue at the beginning of RT was best predicted by the patients' resilience scores, changes of fatigue scores during RT depended on the patients' initial fatigue scores, the decrease in haemoglobin and the patients' experience with RT. CONCLUSIONS: Fatigue appears to be an important problem among cancer patients receiving RT. Resilience proved to powerfully predict the patients' fatigue at the beginning of RT. This result confirms other studies showing resilience to be an important psychological predictor of QoL and coping in cancer patients. The change of fatigue during RT is mainly related to disease- and treatment-related factors. PMID- 18401852 TI - Determination of beta-carboline alkaloids in fruiting bodies of Hygrophorus spp. by liquid chromatography/electrospray ionisation tandem mass spectrometry. AB - The beta-carboline alkaloids harmane (1) and norharmane (2) were isolated from fruiting bodies of Hygrophorus eburneus (Bull.) Fr. as well as brunnein A (3) from Hygrophorus hyacinthinus Quel. (Tricholomataceae, Agaricales) for the first time. Their occurrence within the genus was investigated using liquid chromatography/electrospray ionisation tandem mass spectrometric methods, especially by selected reaction monitoring. Based on these results their chemotaxonomical relevance is discussed. PMID- 18401853 TI - 1H chemical shifts in NMR. Part 27: proton chemical shifts in sulfoxides and sulfones and the magnetic anisotropy, electric field and steric effects of the SO bond. AB - The (1)H chemical shifts of a series of sulfoxide and sulfone compounds in CDCl(3) solvent were obtained from experiment and the literature. These included dialkyl sulfoxides and sulfones (R(2)SO/R(2)SO(2), R = Me, Et, Pr, n-Bu), the cyclic compounds tetramethylene sulfoxide/sulfone, pentamethylene sulfoxide/sulfone and the aromatic compounds p-tolylmethylsulfoxide, dibenzothiopheneoxide/dioxide, E-9-phenanthrylmethylsulfoxide and (E) (Z)-1 methylsulfinyl-2-methylnaphthalene. The spectra of the pentamethylene SO and SO(2) compounds were obtained at -70 degrees C to obtain the spectra from the separate conformers (SO) and from the noninverting ring (SO(2)). This allowed the determination of the substituent chemical shifts (SCS) of the SO and SO(2) functional groups, which were analyzed in terms of the SO bond electric field, magnetic anisotropy and steric effect for long-range protons together with a model (CHARGE8d) for the calculation of the two and three bond effects. After parameterization, the overall root mean square (RMS) error (observed-calculated) for a dataset of 354 (1)H chemical shifts was 0.11 ppm. The anisotropy of the SO bond was found to be very small, supporting the dominant single bond S(+)--O(-) character of this bond. PMID- 18401854 TI - Characterisation of different types of hay by solid-phase micro-extraction-gas chromatography mass spectrometry and multivariate data analysis. AB - The volatile organic compounds of different ground hay samples from Austria, Italy and Switzerland were collected at 50 degrees C on a Supelco Carbowax Divinylbenzene headspace solid-phase microextraction fibre, separated by capillary gas chromatography on an HP5-ms column running a temperature programme and using helium as carrier gas, detected with a mass sensitive detector and studied with principal component analyses after autoscaling selected variables. The analytes, mainly mono- and sesquiterpenes, were able to cluster differences resulting from the site of production. Coumarin can be used to differentiate hay grown north and south of the main chain of the Alps. Acetic acid is appropriate for distinguishing between hay from Kastelruth and Passeier Valley, two South Tyrolean regions. The average linalool content in aftermath is higher than in hay. PMID- 18401855 TI - Distended azygos and hemiazygos vein without interrupted inferior vena cava in a case of agenesis of the ductus venosus. PMID- 18401856 TI - Genetic variability in the mitochondrial serine protease HTRA2 contributes to risk for Parkinson disease. AB - In one genetic study, the high temperature requirement A2 (HTRA2) mitochondrial protein has been associated with increased risk for sporadic Parkinson disease (PD). One missense mutation, p.Gly399Ser, in its C-terminal PDZ domain (from the initial letters of the postsynaptic density 95, PSD-95; discs large; and zonula occludens-1, ZO-1 proteins [Kennedy, 1995]) resulted in defective protease activation, and induced mitochondrial dysfunction when overexpressed in stably transfected cells. Here we examined the contribution of genetic variability in HTRA2 to PD risk in an extended series of 266 Belgian PD patients and 273 control individuals. Mutation analysis identified a novel p.Arg404Trp mutation within the PDZ domain predicted to freeze HTRA2 in an inactive form. Moreover, we identified six patient-specific variants in 5' and 3' regulatory regions that might affect HTRA2 expression as supported by data of luciferase reporter gene analyses. Our study confirms a role of the HTRA2 mitochondrial protein in PD susceptibility through mutations in its functional PDZ domain. In addition, it extends the HTRA2 mutation spectrum to functional variants possibly affecting transcriptional activity. The latter underpins a previously unrecognized role for altered HTRA2 expression as a risk factor relevant to parkinsonian neurodegeneration. PMID- 18401857 TI - Real-time fluorescence-based detection of furanocoumarin photoadducts of DNA. AB - Real-time fluorescence detection systems were adapted to identify DNA adducts formed by photogenotoxic phytochemicals. Two assays were developed: the first was based on quantitative polymerase chain reaction (qPCR) while the second used thermal denaturation and renaturation (D-R). Both assays employed yeast DNA, the fluorescent dye SYBR Green and a real-time PCR thermocycler. The furanocoumarins 8-methoxypsoralen (8-MOP), 5-methoxypsoralen (5-MOP), psoralen, angelicin and imperatorin, and the furanochrome khellin, were tested for adduct forming ability with up to 2 h of UVA light exposure (lambda = 320-400 nm). The known bifunctional compounds, 8-MOP, 5-MOP and psoralen, were inferred to form biadducts here based on both D-R and qPCR assays, as expected from previous research. The known monofunctional compound angelicin was used as a negative control and did not form biadducts based on either assay. Two compounds of unknown functional specificity, imperatorin and khellin, were determined to be positive and negative for biadduct activity, respectively. Detection of biadducts with 8-MOP, 5-MOP, psoralen and imperatorin, but not angelicin or khellin, was further verified by temperature gradient gel electrophoresis. The fluorescence methods improve and expand upon existing assays to monitor DNA adducts. PMID- 18401858 TI - Ion yield improvement for static secondary ion mass spectrometry by use of polyatomic primary ions. AB - Static secondary ion mass spectrometry (S-SIMS) is one of the potentially most powerful and versatile tools for the analysis of surface components at the monolayer level. Current improvements in detection limit (LOD) and molecular specificity rely on the optimisation of the desorption-ionisation (DI) process. As an alternative to monoatomic projectiles, polyatomic primary ion (P.I.) bombardment increases ion yields non-linearly. Common P.I. sources are Ga+ (liquid metal ion gun (LMIG), SF5+ (electron ionisation) and the newer Au(n)+, Bi(n)q+ (both LMIG) and C60+ (electron ionisation) sources. In this study the ion yield improvement obtained by using the newly developed ion sources is assessed. Two dyes (zwitterionic and/or thermolabile polar functionalities on a largely conjugated backbone) were analysed as a thin layer using Ga+, SF5+, C60+, Bi+, Bi3(2+) and Bi5(2+) projectiles under static conditions. The study aims at evaluating the improvement in LOD, useful and characteristic yield and molecular specificity. The corrected total ion count values for the different P.I. sources are compared for different instruments to obtain a rough estimate of the improvements. Furthermore, tentative ionisation and fragmentation schemes are provided to describe the generation of radical and adduct ions. Characteristic ion yields are discussed for the different P.I. sources. An overview of the general appearances of the mass spectra obtained with the different P.I. sources is given to stress the major improvement provided by polyatomic P.I.s in yielding information at higher m/z values. PMID- 18401859 TI - Pyoderma gangrenosum treated with infliximab in inactive ulcerative colitis. PMID- 18401860 TI - Chiral separations in sub- and supercritical fluid chromatography. AB - Sub- and supercritical fluid chromatography (SFC) received more and more attention in pharmaceutical analysis during the last years. Especially for chiral separations, this technique is becoming increasingly popular. This review gives an overview of most chiral separation applications using SFC, covering the literature from 2000 on. PMID- 18401861 TI - Designing supercritical carbon dioxide extraction of rice bran oil that contain oryzanols using response surface methodology. AB - This study examines the supercritical carbon dioxide (SC-CO(2)) extraction of oryzanols contained rice bran oil from powdered rice bran. The extraction efficiencies and concentration factors of oryzanols, free fatty acids and triglycerides in the SC-CO(2) extracts were determined. With top-flow type SC CO(2) extraction the total oil yield was 18.1% and the extraction efficiencies of oryzanols and triglycerides were 88.5 and 91.3% respectively, when 2750 g CO(2 )was consumed during the extraction of 35 g rice bran powder. The concentration factors of oryzanols and triglycerides in SC-CO(2)-extracted oil were higher than in the Soxhlet n-hexane extracted oil. SC-CO(2) extractions indicated that pressure can be used more effectively than temperature to enhance the extraction efficiency and concentration factor of oryzanols. A two-factor central composite scheme of response surface methodology was employed to determine the optimal pressure (300 bar) and temperature (313 K) for increasing the concentration of oryzanols in the SC-CO(2) extracted oil. PMID- 18401862 TI - The advantages of ESI-MS detection in conjunction with HILIC mode separations: Fundamentals and applications. AB - The analysis of highly hydrophilic, ionic, and polar compounds has been performed by HILIC-ESI-MS for the last few years. The use of low aqueous/high polar organic solvent content in HILIC separation mobile phase is almost ideal for ESI-MS detection in many cases, resulting in increased sensitivity. Although the addition of modifiers such as acids or salts is necessary in some circumstances for a good separation, the optimum concentrations used are still highly amenable for ESI-MS analysis, showing few deleterious effects. In this review, the mechanism of HILIC separation and ESI ion generation will be briefly discussed, followed by a summary of method development and applications in several fields of research including pharmaceutical, biomolecular, food, metabolic, and environmental analysis. PMID- 18401863 TI - Quantification of doxazosin in human plasma using hydrophilic interaction liquid chromatography with tandem mass spectrometry. AB - Hydrophilic interaction LC with MS/MS (HILIC-MS/MS) was described as a rapid, sensitive, and selective method for the quantification of doxazosin in human plasma. Doxazosin and cisapride (internal standard) were extracted from human plasma with ethyl acetate at alkaline pH and analyzed on an Atlantis HILIC Silica column with the mobile phase of ACN/ammonium formate (100 mM, pH 4.5) (93:7 v/v). The analytes were detected using an ESI MS/MS in the selective-reaction monitoring mode. The standard curve was linear (r = 0.9994) over the concentration range of 0.2-50 ng/mL. The LOQ for doxazosin was 0.2 ng/mL using 100 microL plasma sample. The CV and relative error for intra- and interassay at four QC levels were 3.7-8.7% and 0.0-9.8%, respectively. The matrix effect for doxazosin and cisapride were practically absent. The recoveries of doxazosin and cisapride were 67.4 and 61.7%, respectively. This method was successfully applied to the pharmacokinetic study of doxazosin in humans. PMID- 18401864 TI - A ruthenium bridge in fullerene-ferrocene arrays: synthesis of [Ru(C60Me5)R(CO)2] (R=C6H4Fc, C identical withCFc) and their charge-transfer properties. AB - New fullerene-ferrocene arrays, [Ru(C(60)Me(5))(C(4)H(6)Fc)(CO)(2)] (Fc=ferrocenyl) and [Ru(C(60)Me(5))(CCFc)(CO)(2)], in which the ruthenium complex functions as a conjugative bridge, were synthesized by the reaction of [Ru(C(60)Me(5))Cl(CO)(2)] with FcC(6)H(4)MgBr and FcCCLi, respectively. These compounds were investigated by electrochemical measurement, single-crystal X-ray structural analysis, and photophysical measurement. Upon photoirradiation, the former compound was converted rapidly into the corresponding triplet state in toluene (tau(singlet)=21 ps), whereas the charge-separated state was predominant in THF (tau(singlet)=10.5 ps; tau(CS)=355 ps). The latter compound, on the other hand, formed the charge-separated state in both toluene and THF (tau(singlet)=3.0 ps; tau(CS)=152 ps). Thus, the structural difference between the phenylene and acetylene bridges in 1 and 2, respectively, was found to change the outcome of the photophysical processes. PMID- 18401865 TI - Grafting BSA onto poly[(L-lactide)-co-carbonate] microspheres by click chemistry. AB - Model protein bovine serum albumin (BSA) was covalently grafted onto poly[(L lactide)-co-carbonate] microsphere surfaces by "click chemistry." The grafting was confirmed by confocal laser scanning microscopy and X-ray photoelectron spectroscopy. The maximum amount of surface-grafted BSA was 45 mg x g(-1). The secondary structure of the grafted BSA was analyzed by FTIR and the results demonstrated that the grafting did not affect protein structure. This strategy can also be used on microspheres prepared from poly(L-lactide)/poly[(L-lactide) co-carbonate] blend materials. PMID- 18401867 TI - Development of electroactive and elastic nanofibers that contain polyaniline and poly(L-lactide-co-epsilon-caprolactone) for the control of cell adhesion. AB - In this work, electrically conductive polyaniline (PAni) doped with camphorsulfonic acid (CPSA) is blended with poly(L-lactide-co-epsilon caprolactone) (PLCL), and then electrospun to prepare uniform nanofibers. The CPSA-PAni/PLCL nanofibers show a smooth fiber structure without coarse lumps or beads and consistent fiber diameters (which range from 100 to 700 nm) even with an increase in the amount of CPSA-PAni (from 0 to 30 wt.-%). However, the elongation at break decreases from 391.54 +/- 9.20% to 207.85 +/- 6.74% when 30% of CPSA-PAni is incorporated. Analysis of the surface of the nanofibers demonstrates the presence of homogeneously blended CPSA-PAni. Most importantly, a four-point probe analysis reveals that electrical properties are maintained in the nanofibers where the conductivity is significantly increased from 0.0015 to 0.0138 S x cm(-1) when the nanofibers are prepared with 30% CPSA-PAni. The cell adhesion tests using human dermal fibroblasts, NIH-3T3 fibroblasts, and C2C12 myoblasts demonstrate significantly higher adhesion on the CPSA-PAni/PLCL nanofibers than pure PLCL nanofibers. In addition, the growth of NIH-3T3 fibroblasts is enhanced under the stimulation of various direct current flows. The CPSA-PAni/PLCL nanofibers with electrically conductive properties may potentially be used as a platform substrate to study the effect of electrical signals on cell activities and to direct desirable cell function for tissue engineering applications. PMID- 18401868 TI - Hydrogen atom release dynamics in radical-radical reactions: saturated vs unsaturated. PMID- 18401866 TI - Release of prostaglandin E(1) from N-(2-hydroxypropyl)methacrylamide copolymer conjugates by bone cells. AB - Bone-targeting N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-PGE(1) conjugates, containing cathepsin K sensitive spacers, were incubated with induced osteoclasts and osteoblasts, their precursors, and control non-skeletal cells. The release of PGE(1) was monitored by an HPLC assay. In both murine and human cell lines, osteoclasts appeared to be the most active cells in the cleavage (PGE(1) release). Incubation with osteoblasts also resulted in fast PGE(1) release, whereas precursor and control cells released PGE(1) with a substantially slower rate than bone cells (apparently through ester bond cleavage). Experiments in the presence of inhibitors revealed that other enzymes, in addition to cathepsin K, were participating in the cleavage of the conjugate. Confocal fluorescence studies exposed internalization of the conjugate by endocytosis with ultimate localization in the lysosomal/endosomal compartment. PMID- 18401873 TI - Organic-inorganic nanohybrids through the direct tailoring of semiconductor nanocrystals with conjugated polymers. AB - Two main synthetic strategies are used to prepare conjugated oligomer- or polymer nanocrystal nanohybrids. In the first strategy ligand exchange is invoked to either replace with a bifunctional ligand, which contains a second functional group for coupling with conjugated oligomers or polymers (COs or CPs), or exchange for the functionalized COs or CPs in a "grafting-onto" process. Alternatively, in the second strategy the nanocrystal (NC) is passivated with functional ligands from which COs or CPs are directly grown in the absence of ligand exchange. The well-defined interface between the COs or CPs and NCs facilitates an efficient charge-transfer between them. PMID- 18401874 TI - Persons with HIV/AIDS face downside of living longer. PMID- 18401875 TI - HIV research opens up potential targets for drugs. PMID- 18401876 TI - Report from the 14th Retrovirus Conference. Treatment options for antiretroviral naive patients. PMID- 18401877 TI - Report from the 14th Retrovirus Conference. The promise of prevention: male circumcision and microbicides. PMID- 18401878 TI - Report from the 14th Retrovirus Conference. Health implications of low CD4-cell counts. PMID- 18401879 TI - Report from the 4th IAS Conference on HIV Pathogenesis, Treatment and Prevention. Maraviroc in treatment-experienced and treatment-naive patients. PMID- 18401880 TI - Can a statin reduce mortality and CV events in elderly CHD patients? PMID- 18401881 TI - Overcoming kinase resistance in chronic myeloid leukemia. AB - Imatinib is a small-molecule inhibitor of BCR-ABL tyrosine kinase activity, with proven efficacy and tolerability. Despite imatinib's activity, the development of resistance, whether BCR-ABL dependent or independent, is a concern. BCR-ABL dependent resistance is commonly a result of mutations in the BCR-ABL gene, which can induce a structural predisposition towards the active conformation of the protein, resulting in a shift in the equilibrium of BCR-ABL from inactive, which imatinib binds, to active, which imatinib is unable to bind. BCR-ABL gene amplification may play a role in the development of imatinib resistance in patients with CML. There are a number of BCR-ABL-independent mechanisms of imatinib resistance, including the efflux protein multidrug resistance protein-1, of which imatinib is a substrate. Another mechanism may be the development of alternative pathways of disease progression, leading to less reliance on BCR-ABL; indeed, the SRC family tyrosine kinases LYN and HCK have been frequently implicated in treatment resistance and progression of CML. Clearly, imatinib resistance requires the development of other treatment options. Dasatinib, with increased binding potency (325-fold greater potency than imatinib for wild-type BCR-ABL), inhibition of both the active and inactive formation of BCR-ABL, and targeting of SRC family kinases, is the only agent approved for the treatment of patients with imatinib-resistant or -intolerant CML and Ph+ ALL. Dasatinib is highly active in all phases of these diseases, and is active in the majority of imatinib-resistant mutations, with the exception of T315I. The development of agents that effectively inhibit T315I mutations suggests that future treatment options will include combination therapy. PMID- 18401882 TI - Weber and Christian of Weber-Christian disease. PMID- 18401883 TI - Abandoning negative marking. PMID- 18401884 TI - Re: Borin JF, Melamud O, Clayman RV. Initial experience with full-length metal stent to relieve malignant ureteral obstruction. J Endourol 2006;20:300-304. PMID- 18401885 TI - Bear attack--A unique fatality in Finland. AB - Fatalities due to animal bites, the vast majority of which are associated with dogs and big cats, are relatively uncommon and rarely described in the literature. Especially rare are fatal bear attacks on humans. We herein present a forensic investigation of a fatal assault, involving numerous bites on a 42-year old man in Finland by an European brown bear (Ursus arctos arctos). PMID- 18401886 TI - Re: Effectiveness of antidepressants. PMID- 18401890 TI - Crystal engineering with urea and thiourea hydrogen-bonding groups. AB - The utilization of N,N'-disubstituted ureas and thioureas as design elements in the synthesis of crystalline organic solids is reviewed. These hydrogen-bonding units are versatile yet predictable building blocks that can be rationally employed in both crystal assembly and functionalization. PMID- 18401891 TI - DNA-templated CMV viral capsid proteins assemble into nanotubes. AB - This communication describes the in vitro assembly of genetically recombinant Cucumber Mosaic Virus (CMV) viral capsid proteins (CPs) into biological nanotubes, several micrometres long yet with a diameter of only approximately 17 nm, triggered by double-stranded DNAs of different lengths. PMID- 18401892 TI - Anion binding vs. sulfonamide deprotonation in functionalised ureas. AB - Sulfonamide groups, commonly used as neutral hydrogen bond donors in a wide variety of anion receptors, deprotonate upon addition of certain basic anionic guests in two simple functionalised ureas. PMID- 18401893 TI - The RNA2-PNA2 hybrid i-motif-a novel RNA-based building block. AB - We report the formation of a hybrid RNA2-PNA2 i-motif comprised of two RNA and two PNA strands based on the sequence specific self assembly of RNA, with potential as a building block for structural RNA nanotechnology. PMID- 18401894 TI - Lithium-promoted hydrogenation of carbon dioxide to formates by heterobimetallic hydridozinc alkoxide clusters. AB - The remarkably distinct reactivity of hydridozinc heterobimetallic cubanes [(HZnOtBu)4-n(thf LiOtBu)n] 1a-1d towards CO2 is reported--the hydride transfer from Zn-H to CO2 is drastically accelerated in the presence of Li ions in 1b-1d which led to the respective metal formate hydrates; the systems are inspiring models for the selective conversion of water gas into formates on lithium promoted ZnO supports. PMID- 18401895 TI - Continuous catalytic Friedel-Crafts acylation in the biphasic medium of an ionic liquid and supercritical carbon dioxide. AB - Immobilisation of catalytically-active metal salts in ionic liquids, with extraction by supercritical carbon dioxide, affords continuous Friedel-Crafts acylation, with in situ-recycling of the catalyst. PMID- 18401896 TI - Activation and reduction of diethyl ether by low valent uranium: formation of the trimetallic, mixed valence uranium oxo species [U(CpRR')(mu-I)2]3(mu3-O) (CpRR' = C5Me5, C5Me4H, C5H4SiMe3). AB - The reaction of UI3 and KCpRR' (CpRR' = pentamethylcyclopentadienyl, trimethylsilylcyclopentadienyl or tetramethylcyclopentadienyl) in diethyl ether results in the two-electron reduction of the solvent to form trimetallic, mixed valence uranium oxo species. PMID- 18401897 TI - Infusion of dye molecules into Red clover necrotic mosaic virus. AB - The Red clover necrotic mosaic virus capsid is utilized to package and release molecules through reversible depletion and re-addition of divalent cations. PMID- 18401898 TI - Health plans target advanced imaging services: cost, quality and safety concerns prompt renewed oversight. AB - Faced with double-digit annual increases in the use of advanced imaging services, health plans are stepping up efforts to manage imaging utilization, maintain imaging quality and ensure patient safety, according to findings from the Center for Studying Health System Change's (HSC) 2007 site visits to 12 nationally representative metropolitan communities. Plan strategies range from informing physicians about evidence-based imaging guidelines to requiring prior authorization of services to credentialing physicians and imaging equipment. Mindful of the physician backlash against managed care in the 1990s, plans are instituting requirements they perceive to be less intrusive and burdensome for physicians. Some physicians, however, view the requirements as administratively onerous and obstacles to patient care. Depending on the experience with imaging, plans may expand utilization management to other services with rapid volume increases. PMID- 18401900 TI - Oral cancer--time for another look? PMID- 18401899 TI - Consumer-directed health plans: mixed employer signals, complex market dynamics. AB - Health plans have expanded consumer-directed health plan (CDHP) product offerings -typically high-deductible health plans coupled with a spending account--and more employers are offering these products to workers, according to findings from the Center for Studying Health System Change's (HSC) 2007 site visits to 12 nationally representative metropolitan communities. In developing CDHPs, health plans are responding to a broader employer strategy to confer more responsibility on workers for their health care costs, lifestyle choices and treatment decisions. CDHP adoption by employers and consumers depends on a range of factors, including product features and employer characteristics, and varies across the 12 communities. While more large employers are introducing CDHPs into health benefit programs, adoption of CDHPs remains modest. Health plans and employers expect CDHP enrollment to grow as employers and employees become more knowledgeable about CDHP features, health plans develop more sophisticated support tools for plan enrollees, and there are more opportunities to learn from early adopters' experiences.early PMID- 18401901 TI - Toronto Academy of Dentistry winter clinic panel discussion on grey market and counterfeit dental materials. PMID- 18401902 TI - Guideline for the early detection of oral cancer in British Columbia 2008. PMID- 18401903 TI - Oral isotretinoin is as effective as a combination of oral isotretinoin and topical anti-acne agents in nodulocystic acne. AB - BACKGROUND: Due to the late introduction of oral isotretinoin, there is only a single report of the use of this drug from India. AIMS: To evaluate the efficacy, acceptability and side effects of oral isotretinoin in Indian conditions and to compare the efficacy of oral isotretinoin monotherapy with that of a combination of oral isotretinoin and topical anti-acne agents. METHODS: Sixty clinically diagnosed patients of nodulocystic acne were divided into two groups of 30 each. Patients in group A were administered oral isotretinoin 20 mg twice a day along with topical clindamycin (1%) and adapalene (0.1%) for 24 weeks. Patients in group B were given only oral isotretinoin. An objective clinical scoring was repeated every four weeks and patients were followed up for six months on completion of treatment. RESULTS: Out of the 60 patients, 50 completed the treatment and 35 completed the follow-up at six months. The mean pretreatment score in group A came down from 12.8 to 1.24 and in group B from 12.4 to 1.48, thus showing a 90.55% and 88% reduction in pretreatment scores respectively. There was no statistical difference in the results obtained from the two groups. Side effects were common but minor in nature. CONCLUSION: Isotretinoin produces gratifying results in patients of nodulocystic acne in Indian conditions. Addition of topical antiacne agents does not alter the final outcome. This addition is well tolerated but requires careful monitoring. PMID- 18401904 TI - Cerebriform elephantiasis of the vulva following tuberculous lymphadenitis. PMID- 18401905 TI - [Clinical case]. PMID- 18401906 TI - [Clinical case]. PMID- 18401908 TI - Wound pain: revisiting a time-honored strategy. PMID- 18401907 TI - [Clinical case]. PMID- 18401909 TI - Wound closure and gradual involution of an infantile hemangioma using a noncontact, low-frequency ultrasound therapy. AB - Facial hemangiomas are benign vascular neoplasms of infancy and childhood. Although most hemangiomas do not result in significant complications, some result in pain, ulceration, infection, and structural abnormalities and can interfere with respiration, vision, and hearing. A 5-month-old infant presented to the author's wound center with a necrotic wound within a large, segmental hemangioma of mixed superficial and deep morphology; her vision was obstructed. The patient's discomfort was indicated by poor feeding and irritability. The hemangioma had been treated with oral corticosteroids and laser therapy. The wound within had been treated with povidone-iodine, moist healing, and topical growth factors. Debridement of necrotic tissue was indicated at presentation; however, bleeding and discomfort were risks. Daily noncontact, low-frequency ultrasound (used for painless debridement of chronic wounds) was initiated to remove necrotic tissue and prepare the wound for grafting. By day 8 of this treatment, the wound bed was covered with healthy-appearing granulation tissue and epithelialization had begun. Additionally, the hemangioma began to involute. Laser treatments were discontinued and the patient was weaned from oral steroids. By week 7, the wound was completely closed and did not require skin grafting. The hemangioma continued to involute. Additional studies are warranted to elucidate the role of noncontact, low-frequency ultrasound in the management of ulcerated hemangioma. PMID- 18401910 TI - Physiological studies in tropical savannas and seasonally dry forests. Preface. PMID- 18401911 TI - Continuing competence: one state's efforts. PMID- 18401912 TI - [Organization of Hospitals for Cancer Treatment. Cancer Act and Council for National Cancer Strategy]. PMID- 18401913 TI - Pulmonary artery configuration after the Norwood procedure for hypoplastic left heart syndrome. PMID- 18401914 TI - [The decline of rosiglitazone]. PMID- 18401915 TI - [Physicians' outlook on mankind]. PMID- 18401916 TI - [Musculoskeletal tumor]. PMID- 18401917 TI - [Proteomic analysis for connective and soft tissue neoplasms]. PMID- 18401918 TI - [Molecular targeting for treatment of connective and soft tissue neoplasms]. PMID- 18401919 TI - [Current topics of chemotherapy for sarcoma and bone neoplasms]. PMID- 18401920 TI - [Topic of minimally invasive surgery for connective and soft tissue neoplasms]. PMID- 18401921 TI - Evidence-based animal care: new contributions to our knowledge base and the need for more. PMID- 18401922 TI - Position of the American Dietetic Association: nutrition and lifestyle for a healthy pregnancy outcome. AB - It is the position of the American Dietetic Association that women of child bearing ages should maintain good nutritional status through a lifestyle that optimizes maternal health and reduces the risk of birth defects, suboptimal fetal growth and development, and chronic health problems in their children. The key components of a health-promoting lifestyle during pregnancy include appropriate weight gain; appropriate physical activity; consumption of a variety of foods in accordance with the Dietary Guidelines for Americans 2005; appropriate and timely vitamin and mineral supplementation; avoidance of alcohol, tobacco, and other harmful substances; and safe food handling. Pregnant women with inappropriate weight gain, hyperemesis, poor dietary patterns, phenylketonuria, certain chronic health problems, or a history of substance abuse should be referred to a registered dietitian for medical nutrition therapy. Prenatal weight gain within the Institute of Medicine recommended ranges has been associated with better pregnancy outcomes. Most pregnant women need 2,200 to 2,900 kcal a day, but prepregnancy body mass index, rate of weight gain, maternal age, and appetite must be considered when tailoring this recommendation to the individual. The consumption of more food to meet energy needs, and the increased absorption and efficiency of nutrient utilization that occurs in pregnancy, are generally adequate to meet the needs for most nutrients. However, vitamin and mineral supplementation is appropriate for some nutrients and situations. This position paper also includes recommendations pertaining to use of alcohol, tobacco, caffeine, and illicit drugs. PMID- 18401923 TI - Impacts of abstinence education on teen sexual activity, risk of pregnancy, and risk of sexually transmitted diseases. AB - This paper examines the impacts of four abstinence-only education programs on adolescent sexual activity and risks of pregnancy and sexually transmitted diseases (STDs). Based on an experimental design, the impact analysis uses survey data collected in 2005 and early 2006 from more than 2,000 teens who had been randomly assigned to either a program group that was eligible to participate in one of the four programs or a control group that was not. The findings show no significant impact on teen sexual activity, no differences in rates of unprotected sex, and some impacts on knowledge of STDs and perceived effectiveness of condoms and birth control pills PMID- 18401924 TI - Reassessing the WIC effect: evidence from the Pregnancy Nutrition Surveillance System. AB - Recent analyses differ on how effective the Special Supplemental Nutrition Program for Women, Infants and Children (WIC) is at improving infant health. We use data from nine states that participate in the Pregnancy Nutrition Surveillance System to address limitations in previous work. With information on the mother's timing of WIC enrollment, we test whether greater exposure to WIC is associated with less smoking, improved weight gain during pregnancy, better birth outcomes, and greater likelihood of breastfeeding. Our results suggest that much of the often reported association between WIC and lower rates of preterm birth is likely spurious, the result of gestational age bias. We find modest effects of WIC on fetal growth, inconsistent associations between WIC and smoking, limited associations with gestational weight gain, and some relationship with breastfeeding. A WIC effect exists, but on fewer margins and with less impact than has been claimed by policy analysts and advocates. PMID- 18401925 TI - Sex matters. PMID- 18401926 TI - Mandating HPV vaccination: what are the arguments? PMID- 18401927 TI - Working in healthcare today: what would Jesus do? PMID- 18401928 TI - Healthcare missions: mobilizing communities for health. PMID- 18401929 TI - Faith integration in education: how iron sharpens iron. PMID- 18401930 TI - A special bond of life: one educator's story. PMID- 18401931 TI - A begining look at the spiritual practices of older adults. PMID- 18401932 TI - Shed or you're dead. PMID- 18401933 TI - The "5 R's" of sharing the gospel. PMID- 18401934 TI - Measures of success. PMID- 18401935 TI - The power of the rugged cross. PMID- 18401936 TI - How can I respond to hard questions? PMID- 18401937 TI - Child oral health challenges. PMID- 18401938 TI - The regulation of the dental profession in New Zealand. AB - Professions have developed and evolved in response to many different societal pressures. The dental profession in New Zealand is no exception, and (in particular) has been influenced by--and has influenced--changing regulatory environments since the nineteenth century. The Health Practitioners Competence Assurance Act (2003) is the latest Act to regulate dentistry, and has many details that will affect the practice of dentistry in New Zealand and challenge the dental profession to respond in new ways. PMID- 18401939 TI - The provision of dental treatment for children under general anaesthesia. PMID- 18401940 TI - Changes in the New Zealand dentist workforce over a nine-year period. AB - OBJECTIVE: To identify and describe trends in the NZ dentist workforce over a nine-year period. METHODS: Data from the 1997, 2001 and 2005 DCNZ workforce questionnaires were collated and analysed using both cross-sectional and longitudinal methods. RESULTS: 2088 dentists completed the workforce questionnaire in at least one of these years, and 1026 (49.1%) did so in all three of them. Substantial changes occurred in the NZ dentist workforce over the observation period. The proportion of women and overseas-trained dentists in the workforce continued to increase, along with the mean age of dentists. There was a large decrease in the number of hours worked per week by specialist dentists (who still worked more hours than dentists), but there was no clear trend for general dentists. Increasing numbers of dentists appear to be opting for part-time work; the main reasons for doing so included personal choice, parental responsibilities, and semi-retirement. There was a trend for fewer dentists to be working in small towns, mainly through dentists leaving (or retiring) and not being replaced, rather than dentists moving from towns to the cities. Rural dentists and female practitioners were less likely to achieve the continuing professional development (CPD) target than their colleagues. In 1997, fewer overseas-qualified dentists (than NZ graduates) were self-employed and had undertaken 20 hours CPD, and a larger proportion worked in small towns. However, these differences were not evident in the later years. CONCLUSION: The New Zealand dental workforce continues to change, particularly with respect to gender mix, age, work circumstances and CPD activities. PMID- 18401941 TI - Changes in serum leptin, insulin, androstenedione and luteinizing hormone during ovarian cycle in the bat, Taphozous longimanus. AB - Detailed reproductive pattern and associated endocrine characteristics have been documented in only a few species of order Chiroptera. The aim of the present study was to examine the changes in body weight, serum insulin, leptin, androstenedione and luteinizing hormone (LH) concentrations during annual ovarian cycle in the sheath-tailed bat, Taphozous longimanus. Bats were sampled over three years. Leptin, a satiety hormone produced primarily by adipose tissue, provides information to feeding center of the brain about nutritional status, fat mass, appetite and energy expenditure. The circulating concentration of leptin begins to increase from October and attains a peak in December. The peak serum leptin concentration coincides with body weight in November before winter dormancy in December. The serum leptin levels dissociate from body weight during December. The other peaks of serum leptin levels coincide with late stages of the two successive pregnancies. The serum insulin concentration begins to increase from September and attains a peak during December. The insulin concentration remains low from January to August. The circulating androstenedione concentration begins to increase in October, reaching a peak in December. This increase in androstenedione concentration correlated with the period of heavy accumulation of abdominal fat and increase in body weight. There was a sharp decline in androstenedione concentration and body weight in January. The serum LH shows peaks, in November, coinciding with the peaked body weight, the other peaks in January and May, coinciding with ovulation for the two successive pregnancies. The high leptin and insulin levels might be responsible for the maintenance of reproductive response and gonadal function during adverse environmental condition in the winter, while high androstenedione, and associated body weight along with LH might be responsible for maintaining basal gonadal function. We conclude that high leptin, androstenedione and insulin serve, as signal for the reproductive functions in that sufficient body fat are available to meet the caloric demands and maintain normal function during adverse winter conditions. PMID- 18401942 TI - Effects of dexamethasone on insulin receptor in aging. AB - The aim of this study was to examine the effects of dexamethasone (Dex) on functional properties of the rat insulin receptor (IR). Male Mill Hill hooded rats, 3, 6, 12, 18 and 21 months old, were injected with Dex (4 mg/kg) and rat liver and erythrocytes were used for experiments 18 h after Dex administration. Treatment with Dex lowered the specific binding (SB) of insulin (INS) in the liver of 3- and 18-month-old rats and concentration of INS binding sites (N1, N2) and the dissociation constant of low-affinity binding sites (Kd2) in the liver of 6- and 18-month-old rats. In addition, Dex treatment lowered the liver IR protein level in all analyzed groups, except 21-month-old rats where it remained unchanged, but raised the IR mRNA level in 18-month-old rats. In erythrocytes, treatment with Dex decreased SB and Kd2 (in animals 3 and 6 months old) and N1 (in ones 3 and 18 months old). Following Dex treatment, the INS plasma level increased (in rats 3, 18 and 21 months old), while glucose (Glu) concentration increased in 3 and 12 months old, but decreased in 6- and 21-month-old rats. In summary, Dex exerts the strongest effect on the erythrocyte IR of 3- and 6-month old rats and the hepatic IR of 18-month-old rats. IR in both tissues is almost insensitive to Dex in 12- and 21-month-old rats. The pattern of age-related changes of IR induced by Dex does not correlate with changes of plasma Glu and INS. PMID- 18401943 TI - Acetylcholinesterase, catalase and glutathione S-transferase activity in beet armyworm (Spodoptera exigua) exposed to nickel and/or diazinon. AB - The effect of single and combined action of nickel and pesticide (diazinon) on enzymes activity (glutathione S-transferase, catalase and acetylcholinesterase) in the digestive tract, body wall and fat body as well as basic growth parameters (life span and body mass) of Spodoptera exigua were investigated under laboratory conditions. The experiment was carried out on two nickel treated groups [300 (NiI) and 900 (NiII) mg Ni kg(-1) dry weight of the culture medium] and a control group. The results showed that mortality of caterpillars in NiII group was higher (51.1%) when compare with the controls. The body mass of the caterpillars in the NiI group was higher by 20% than in the control group, and the body mass of the pupae in the NiII group was lower by 22% than in the control group. Exposure to nickel influenced AChE, GST and catalase activity in the body wall (increase up to 66%) and GI tract, while in the fat body the above-mentioned activity remained unchanged. The pesticide application caused a strong, about 70% reduction in AChE activity in GI tract, while in case of GST activity pesticide treatment resulted in multidirectional response depending on the organ. Nickel pre-treatment affects the susceptibility to pesticide, which is manifested in a lower activity of GST and catalase in the fat body (from 26 to 36%), when compare with the other experimental groups. PMID- 18401944 TI - The effect of lead on fitness components and developmental stability in Drosophila subobscura. AB - We analyzed the developmental time, egg-to-adult viability, and developmental stability (fluctuating wing size asymmetry) in Drosophila subobscura, maintained for six generations on different concentrations of lead. Development time is significantly affected by generation and lead concentration, but interaction of these factors is not a significant source of variability for this fitness component. Generation and the interaction generation x concentration of lead significantly affect egg-to-adult viability. Levene's test of heterogeneity of variance showed that variability of FA is not significant in any of the samples. Within both lead concentrations females showed significantly higher FA indices for the wing width than males. Within sexes, a significantly higher FA was found only in females for wing width FA between the control and the lower concentration of lead. The results show that if strong relationship between FA and the studied fitness components exists, it results in a stronger selection of unstable genotypes under lead as a stress factor and, consequently, FA needs to be used with caution as a biomarker in natural populations under environmental stress. PMID- 18401945 TI - Antioxidative and metabolic responses to extended cold exposure in rats. AB - In this work, we investigated whether extended cold exposure increases oxidative damage and susceptibility to oxidants of rat liver, heart, kidney and lung which are metabolically active tissues. Moreover in this study the effect of cold stress on some of the lipid metabolic mediators were studied in rat experimental model. Male albino Sprague-Dawley rats were randomly divided into two groups: The control group (n=12) and the cold-stress group (n=12). Tissue superoxide dismutase (SOD), catalase (CAT), glutathion S-transferase (GST) and glutathion reductase (GR) activities and glutathion (GSH) were measured using standard protocols. The biochemical analyses for total lipid, cholesterol, trigliceride, HDL, VLDL and LDL were done on autoanalyzer. In cold-stress groups SOD activity was decreased in the lung whereas it increased in the heart and kidney. CAT activity was significantly decreased (except liver) in all the tissues in treated rats. GST activity of cold-induced rats increased in liver and heart while decreased in the lung. GR activity was significantly decreased (except in liver) in all the tissues in cold-stressed rats. GSH level was significantly increased in the heart but decreased in the lung of animals exposed to cold when compared to controls. It was found that among the groups trigliceride, total lipid, HDL and VLDL parameters varied significantly but cholesterol and LDL had no significant variance. In this study, we found that exposure of extended (48 h) cold (8 degrees C) caused changes both in the antioxidant defense system (as tissue and enzyme specific) and serum lipoprotein profiles in rats. PMID- 18401946 TI - Impact of 864 MHz or 935 MHz radiofrequency microwave radiation on the basic growth parameters of V79 cell line. AB - The aim of this study was to evaluate and compare the influence of 864 MHz and 935 MHz radiofrequency/microwave (RF/MW) fields on the growth, colony-forming ability, and viability of V79 cells (continuous line). Cell samples with 1 x 10(4) V79 cells each, were exposed to continuous wave frequencies of 864 MHz and 935 MHz for 1, 2 and 3 hours. Exposed samples were matched with unexposed control samples. Specific absorption rate (SAR) was 0.08 W/kg for the 864 MHz or 0.12 W/kg for the 935 MHz field. Cell growth and viability were determined by counting cells every day for five days after exposure. Colony-forming ability was assessed by counting colonies seven days after exposure. The growth of the 864 MHz irradiated cells was significant after two- and three-hour exposure 72 hours after irradiation (p < 0.05). The similar was observed 72 hours after exposure for cells exposed to 935 MHz microwaves for three hours (p <0.05). Colony-forming ability and cell viability in V79 cells exposed to 864 MHz or 935 MHz microwaves did not significantly differ from control cells. The two applied RF/MW fields showed similar effects on the growth, colony-forming ability and viability of V79 cells. Cell growth impact was time-dependent for both fields. PMID- 18401947 TI - Selection of salt tolerant plants of Nicotiana tabacum L. through in vitro and its biochemical characterization. AB - Sodium chloride tolerant organogenic callus lines of Nicotiana tabacum were developed in vitro on Murashige and Skoog [16] medium supplemented with BA, IAA and different concentration of NaCl. The maximum shoot bud regeneration was achieved from both tolerant and non-tolerant calluses on MS medium supplemented with 1.0 mg/l BA, 0.1 mg/l IAA with or without NaCl within 4 weeks of culture. Standard growth parameters such as fresh weight and dry weight of organogenic callus, growth tolerant index and enzyme activity (peroxidase and catalase) were used as indicators of salt tolerance. The growth tolerance index in the 4-week after the beginning of treatments yielded significant differences among the non tolerant and tolerant organogenic callus lines. The regenerated shoots were rooted on half-strength MS basal salts supplemented with 2% sucrose but devoid of growth regulator. The regenerated plants from tolerant callus lines were capable of growing in vitro in presence of 175 mM NaCl. SDS-PAGE profile showed that the progenies derived from tolerant sources were tolerant to salt. This investigation may help in the selection and characterization of salt tolerance in plant improvement programme. PMID- 18401948 TI - Effects of polyamines and polyamine biosynthetic inhibitors on mitotic activity of Allium cepa root tips. AB - The genotoxic and cytotoxic effects of exogenous polyamines (PAs), putrescine (Put), spermidine (Spd), spermine (Spm) and PA biosynthetic inhibitors, alpha difluoromethylornithine (DFMO), cyclohexilamine (CHA), methylglioxal bis (guanylhydrazone) (MGBG) were investigated in the root meristems of Allium cepa L. The reduction of mitotic index and the induction of chromosomal aberrations such as bridges, stickiness, c-mitotic anaphases, micronuclei, endoredupliction by PAs and PA biosynthetic inhibitors were observed and these were used as evidence of genotoxicity and cytotoxicity. PMID- 18401949 TI - Leaf width, nrDNA and cpDNA its sequence variation within central European Bulbocodium vernum and B. versicolor (Colchicaceae) populations: are there really two taxa? AB - The taxonomy of the genus Bulbocodium, in which two European species, a smaller eastern (B. versicolor) and a more robust western (B. vernum) are included, has been controversial since the description of the eastern species in 1821. Nuclear encoded ribosomal DNA ITS1 and the entire chloroplast DNA ITS were sequenced from several European populations, from France to the Ukraine, and the leaf width of mature living individuals was measured and analysed by ANOVA and Tukey-test. Although the studied DNA regions proved to be invariable, leaf width shows extreme variability. We found no correlation between the leaf size of the individuals and the geographical position of the populations, and in addition, the sequenced DNA regions showed total uniformity. Thus, our results do not support the division of the genus Bulbocodium into two taxa, at least in the sampled area. The formerly described size variants can be treated taxonomically at the forma level. PMID- 18401950 TI - Influence of phosphate and ammonia on the growth, exopolysaccharide production and symbiosis of Rhizobium leguminosarum bv. trifolii TA1 with clover (Trifolium pratense). AB - The Rhizobium-legume interaction is sensitive to a number of environmental factors, among which phosphate (Pi) and ammonium availability are the most important. We investigated the effect of Pi and ammonia concentration on exopolysaccharide production and symbiosis Trifolium pratense with Rhizobium leguminosarum bv. trifolii TA1 (RtTA1). The optimal Pi concentration in the bacterial growth medium for RtTA1 growth and exopolysaccharide production was in the range from 0.9 mM to 8.1 mM. Independently of Pi concentration, ammonium (NH4Cl) concentration above 8.1 mM in the culture medium significantly decreased EPS production, indicating a regulatory role of this nutrient on the EPS production in the RtTA1 strain. Pi availability has a beneficial effect on both partners of symbiosis. Pi concentration in the plant medium in the range from 1.7 mM to 5 mM was optimal for nodule formation, nodule occupancy and nitrogen fixation ability. Despite of T. pratense cv. Bryza tolerance on high Pi concentration, 20 mM Pi occurs to be nearly phytotoxic, which negatively affects almost all symbiotic parameters. Large amounts of starch were accumulated in the nodules formed by clover grown on medium containing high Pi concentration. PMID- 18401951 TI - Failure of oral E. coli O83 lipopolysaccharide to influence intestinal morphology and cell proliferation in rats: short communication. AB - It is known that bacterial lipopolysaccharide (LPS) is not absorbed from the gastrointestinal tract in significant quantities. However, the question remained whether oral LPS modified the structure or function of the gut. In the present experiment Escherichia coli 083 LPS was administered to growing rats in repeated oral doses of 400 mg/kg body weight (b. w.), every 8 h. After three days of treatment, morphometric and histochemical examinations of the small intestine did not show significant differences between treated and control rats. It is concluded that repeated oral administration of high doses of E. coli 083 LPS had no demonstrable effect on intestinal structure and cell proliferation in a rat model. PMID- 18401952 TI - Salmonella Genomic Island 1 (SGI1) and genetic characteristics of animal and food isolates of Salmonella typhimurium DT104 in Hungary. AB - To study the genetic characteristics of DT104 strains of Salmonella Typhimurium and the prevalence of Salmonella Genomic Island (SGI1) in Hungary, 140 recent Salmonella strains of food and animal origin were examined. For the first time in Hungary, the SGI1 was found in 17 out of 59 S. Typhimurium isolates (all proven to be DT104 phage type). These 17 strains were then subtyped by pulsed-field gel electrophoresis (PFGE) into 6 pulsotypes which were less correlated with the geographic origin than with the animal species of origin. PMID- 18401953 TI - Distribution and genetic relatedness of vancomycin-resistant enterococci (VRE) isolated from healthy slaughtered chickens in Hungary from 2001 to 2004. AB - The presence of the vanA gene was determined in enterococci from healthy poultry, originating from the Hungarian resistance monitoring system between 2001 and 2004. Enterococci (n = 562) were collected from intestinal samples of slaughtered broiler chickens. The presence of van genes was detected by polymerase chain reaction (PCR). The vancomycin-resistant enterococcus (VRE) strains carried only the vanA gene. Genus- and species-level identification of the vanA gene carrier strains was carried out by PCR using specific primers. In 2001, 25 out of the 289 isolated strains (8.6%) were vanA carriers (1 Enterococcus mundtii, 13 E. durans and 11 E.faecium). In 2002 (n = 87), 20 (23%) strains were vanA positive (11 E. durans and 9 E. faecium). In 2003 and 2004, none of the strains (n = 95 and 91, respectively) were positive for the most common van genes. In 2003, there was only one strain for which higher minimum inhibitory concentrations (MIC) of vancomycin (4 mg/L) and teicoplanin (8 mg/L) were found. In 2004 there were three strains for which the MIC of vancomycin was 8 mg/L, and 2 strains and 1 strain with teicoplanin MICs of 4 mg/L and 8 mg/L, respectively. The potential similarity of these strains was studied by pulsed-field gel electrophoresis (PFGE). The VRE strains were not closely related to one another. The annual data of vancomycin resistance indicate an association between the recovery of vancomycin-resistant enterococci and the use of avoparcin in animal feeds. This study indicates that with the reduced use of antibiotics in food animals, it is possible to decrease the rate of resistant bacteria. Although the use of avoparcin had been banned in 1998, the VRE strains disappeared only five years later. PMID- 18401954 TI - Atrophic rhinitis vaccine composition triggers different serological profiles that do not correlate with protection. AB - Atrophic rhinitis (AR) is a widespread and economically important disease of swine caused by Bordetella bronchiseptica and Pasteurella multocida. It can be controlled by vaccination. This study investigates the effect of altering the composition (adjuvants and/or addition of formalin-inactivated P. multocida toxin, fPMT) of conventional vaccines on the serological profile and on protection against AR in swine. A significantly higher B. bronchiseptica specific antibody titre was detected for vaccines with novel immunostimulants, the best being Montanide IMS 1313 (1:630 compared to 1:274 obtained with alum). The highest B. bronchiseptica antibody titre was demonstrated for a combination of B. bronchiseptica--fPMT, while PMT antibody titre was highest for monovalent fPMT (both adjuvanted with IMS 1313). The AR-specific antibodies were transmitted from dams to their offspring in similar titres and with the same hierarchy of effectiveness. After a B. bronchiseptica--P. multocida bacterial challenge, piglets from dams vaccinated with fPMT combined with B. bronchiseptica or B. bronchiseptica--P. multocida bacterins showed the lowest nasal lesions scores (4.5 and 3.2, respectively, out of a possible maximum score of 18). These combinations, both of which were adjuvanted with IMS 1313, gave the best protection against experimentally induced AR. Our results show that the adjuvant and the antigen composition of the vaccine strongly affect seroconversion, and that the AR-specific antibody titre does not necessarily correlate with the degree of protection. PMID- 18401955 TI - Effects of high dietary molybdenum in rabbits. AB - To study the effects of high dietary molybdenum (Mo) content, rabbits were fed with commercial pellets and carrots containing 39 mg Mo/kg dry matter (DM) [Experiment 1] and with a commercial diet supplemented with 40 mg Mo/kg DM [Experiment 2] for 14 days. The high dietary Mo contents failed to reduce the growth performance of rabbits. Moreover, supplemental Mo given in a dose of 40 mg/kg non-significantly decreased the apparent digestibility of crude protein (CP) and crude fibre (CF) compared to the control (73.63 +/- 2.49 and 18.56 +/- 5.10 vs. 74.31 +/- 3.03 and 21.38 +/- 6.48, respectively). Molybdenum ingested with feeds was mainly excreted (57%) via the urine. The highest Mo levels were found in kidney and liver samples (3.464 +/- 0.872; 5.27 +/- 0.95 mg/kg DM [Experiment 1] and 1.878 +/- 0.283; 1.62 +/- 0.16 mg/kg DM [Experiment 2], respectively), and Mo could also be detected in limb meat (0.336 +/- 0.205 mg/kg DM). It was stated that the testes were more sensitive to Mo exposure than the female reproductive organs because the number of germ cells was reduced. Due to the high dietary Mo intake free radicals could be generated, resulting in a marked increase of creatine kinase (CK) activity. PMID- 18401956 TI - Effect of various dietary fat supplementations on liver lipid and glycogen of high-yielding dairy cows in the peripartal period. AB - In a model experiment, Holstein-Friesian dairy cows were fed on a corn-silage based diet supplemented with 11.75 MJ NE1 per day of calcium soaps of palm oil fatty acids (CAS) or hydrogenated triglyceride (HTG) or without fat supplementation (control). All diets were fed to the cows over a period from 21 +/- 3 days (d) prior to the expected calving to d 100 +/- 5 postpartum. On d 25 (basal sample) and d 14 prepartum as well as on d 5 and 25 postpartum liver samples were collected by percutaneous biopsy. Total lipid content, fatty acid composition and glycogen of liver tissues were determined. At d 5 postpartum, both control and CAS cows had higher liver lipid (P < 0.05) and lower glycogen (P < 0.05) concentrations than cows in the HTG group. No significant (P < 0.05) differences were detected in liver fat content among the groups at d 14 prepartum or d 25 postpartum. The glycogen concentration slightly decreased in the liver of cows in each treatment group from d 14 prepartum to d 5 postpartum; however, this decrease was more intensive in both the control and CAS groups than in the HTG group. The variations in liver lipid concentrations were accompanied by significant changes in the proportion of C16:0, C16:1n-7, C18:0, C18:1n-9, C18:2n 6 and C20:4n-6 fatty acids in the liver lipids. The results show that HTG supplementation exerted more advantageous effects on liver lipid and glycogen metabolism than did CAS supplementation. PMID- 18401957 TI - Further evidence for the involvement of anatomical parameters of the cardiopulmonary system in the development of ascites syndrome in broiler chickens. AB - Eggs from a broiler line were incubated at two different altitudes and hatched. Relative heart and lung weights, volumes of the heart, lung and thoracic cavity, incidence of right ventricular hypertrophy and ascites, and related physiological parameters were followed in the day-old chickens hatched from the above eggs. Lung and heart weights as a percentage of body weight, lung and heart volumes relative to the volume of the thoracic cavity after removing the heart and lungs were higher in chickens hatched at high altitude. Additionally, embryonic triiodothyronine (T3) and thyroxine (T4) levels relative to cardiopulmonary parameters were higher in day-old chickens that hatched at high altitude as compared with chickens hatched at low altitude. This was associated with a lower incidence of right ventricular hypertrophy and ascites in chickens hatched at high altitude. Our data indicate that chronic hypoxia interacting with the endogenous functions of embryos during embryonic development at high altitude, as adaptation mechanisms, changed the developmental trajectories of cardiopulmonary parameters in postnatal chickens. This important development facilitates an increase in the gas exchange area in broiler chickens, thus lowering their susceptibility to pulmonary hypertension and ascites. PMID- 18401958 TI - Seroprevalence of Toxoplasma gondii and Neospora caninum infection of cats in Hungary. AB - Blood samples were collected from 330 cats in Hungary in order to evaluate their seroconversion to Toxoplasma gondii and Neospora caninum using the indirect fluorescent antibody test (IFAT). The overall prevalence of toxoplasmosis was 47.6%, the prevalence being 22.4% among urban, 50% among suburban and 61.3% among rural animals. Significantly more cats had high IFAT titres (1:640 to 1:5120) in the countryside. Female cats were more frequently infected with T. gondii than males (53.3% vs. 39.3%), and seropositivity increased with the age of animals. The prevalence (0.6%) and titre (1:40) of antibodies to N. caninum was low. Sixty two cats were also screened for seroconversion to feline infectious peritonitis (FIP) virus. Higher titres to T. gondii were more frequently detected among FIP positive cats, but this difference was non-significant due to the small number of cats with concurrent infection. PMID- 18401959 TI - Application of anti-BCG antibody for rapid immunohistochemical detection of bacteria, fungi and protozoa in formalin-fixed paraffin-embedded tissue samples. AB - The applicability of an anti-Mycobacterium bovis (BCG) antibody-based immunohistochemistry (IHC) procedure was investigated using everyday veterinary pathological samples collected from 13 different animal species. Fifty-one formalin-fixed and paraffin-embedded tissue samples were selected for this study. Forty, 4 and 7 tissue samples contained different species of bacteria, fungi and protozoa, respectively. Three serial sections were prepared in each case. Two sections were pre-treated with enzyme and heat, respectively, while the last section was not pre-treated. In seven cases the sensitivity of histochemical staining (HSM), IHC and bacteriological culture were compared. Heating of the sections in a microwave oven was the most effective method in the case of almost all pathogens used. Strong or moderate positive reactions were observed for 26 bacterial species, all fungal and 2 protozoal species, while weak reactions occurred for 2 bacterial and 1 protozoal species. Only 4 protozoal and 12 bacterial species, including Leptospira and all the five Mycoplasma species examined, showed no reaction in this test. IHC had almost the same sensitivity as bacteriological culture and was more sensitive than HSM. The IHC method presented here should be preferred to HSM as a general screening tool in cases where pathological lesions suspicious for infections are evident and no microorganism can be cultured in vitro or only formalin-fixed tissue samples are available for the laboratory examination. PMID- 18401960 TI - Proliferative and necrotising pneumonia and severe vascular lesions in pigs naturally infected with porcine circovirus type 2. AB - Severe disease induced by porcine circovirus type 2 (PCV2) was observed in three pigs originating from a large herd affected by respiratory and digestive signs as well as wasting. Proliferative and necrotising pneumonia (PNP) was diagnosed in two animals, while severe acute interstitial pneumonia characterised by the presence of abundant hyaline membrane in the alveoli and fibrin in the bronchioles was found in one pig. In all cases, large amounts of PCV2 antigen were found in each tissue sample collected from the lungs and mediastinal lymph nodes. Neither porcine reproductive and respiratory syndrome virus (PRRSV) nor swine influenza virus (SIV) was detected, and no bacteria could be cultured in any of the cases. Vascular lesions, e.g. degeneration of endothelial cells, perivascular and intramural oedema, fibrinoid necrosis, vasculitis, perivasculitis, and vascular thrombi were observed in all cases, associated with the presence of PCV2 antigen. The viral antigen was present in the intravascular mononuclear cells, endothelial cells, myocytes and infiltrating inflammatory cells in lymph and blood vessels. In one case, obliterating thrombi in the lymph and blood vessels were directly connected to areas of tissue necrosis and were associated with abundant PCV2 antigen. The results further suggest the causative role of PCV2 infection in PNP, and the importance of the vascular system in the pathogenesis of PCV2-associated diseases of swine. PMID- 18401961 TI - Feline vaccine-associated fibrosarcoma induced by aluminium compound in two cats: short communication. AB - Two cases of feline vaccine-associated fibrosarcoma (FVAF) are reported. The excised tumours were both characterised as well circumscribed, subcutaneous, firm and white with central necrosis. Histopathologically, they consisted of well differentiated and variably sized and shaped anaplastic cells, characterised by marked nuclear and cellular pleomorphism including giant cells. The mitotic activity was low. Aluminium was demonstrated in the central necrosis and giant cells. Neoplastic cells were positive for vimentin and negative for desmin and cytokeratin. The presence of feline sarcoma virus and feline immunodeficiency virus could not be detected by PCR in either case. PMID- 18401962 TI - Immunohistochemical distribution of alpha B-crystallin in the cerebellum of dogs infected with canine distemper virus. AB - The cerebella of 12 dogs infected with canine distemper virus (CDV) and those of three normal dogs were examined. The avidin-biotin-peroxidase complex technique was used to detect alphaB-crystallin (alphaB-c) immunoreactivity and immunolocalisation of the CDV antigen. CDV antigens, immunopositive astrocytes, oligodendrocytes and granular neurons were seen in both the white and grey matter of the infected dogs. In the controls, alphaB-c immunopositive glial cells were seen in the white matter and around the Purkinje cells. In dogs with distemper, alphaB-c immunoreactivity was not observed in some of the glial cells around the Purkinje cells. A significant negative correlation of P < 0.01 level was found between areas of severe demyelination and the number of alphaB-c immunopositive cells in dogs infected with CDV. Such correlation was not observed between mild and moderate demyelinating areas and alphaB-c immunostaining. The alphaB crystallin/ total number of cells ratio was found to be significant in severely affected demyelinating areas (P < 0.05). These data indicate that there was a relationship between the degrees of CDV associated with demyelination and the level of alphaB-c expression in the glial cells. PMID- 18401963 TI - Melengestrol acetate as a tool for inducing early ovulation in transitional mares. AB - The efficacy of melengestrol acetate (MGA) to shorten the vernal transition of mares by synchronising and accelerating the first ovulation of the year after 60 days of phototherapy was determined by ultrasonographic monitoring. Sixteen mares in late transition were fed two doses of MGA (150 mg/mare/day and 100 mg/mare/day, respectively) for 10 days. A luteolytic dose of prostaglandin was administered to each mare one day after the end of MGA treatment. The presence and duration of oestrus, follicular growth, uterine oedema and presence of ovulation were monitored by ultrasonography and the cervical tone was evaluated by rectal palpation. Ovulation was detected in 87.5% of the mares treated with 150 mg MGA/mare/day for 10 days, and in 62.5% of the mares receiving 100 mg MGA/mare/day for 10 days. This was statistically different (P = 0.03) from the untreated control mares having an ovulation rate of 20%. Mares that received 150 mg MGA/day for 10 days had a mean treatment to ovulation interval of 13.1 +/- 5.97 days after the end of treatment, while mares that received 100 mg MGA/day for 10 days had a mean of 25.6 +/- 10.50 days (P = 0.01) to ovulation. These results suggest that MGA can be used for synchronising and hastening the first ovulation of the year in mares. PMID- 18401964 TI - Acrosomal and viability status of bovine spermatozoa evaluated by two staining methods. AB - Artificial insemination with frozen-thawed spermatozoa is commonly used in cattle breeding. A simple and fast procedure is needed for routine evaluation of the acrosomal status of frozen-thawed bovine sperm. Therefore, the purpose of this study was to test two staining procedures used to determine the viability and integrity of acrosome of frozen-thawed bovine spermatozoa. Double staining and Hoechst/FITC-Pisum sativum agglutinin (FITC-PSA) labelling were tested for evaluating the viability and acrosome reaction induced by calcium ionophore of bull spermatozoa. In our experiments no significant differences were detected in the frequency of acrosome-reacted sperm either by double staining (37.98%) or by FITC-PSA labelling (39.33%). The viability of sperm stained by the double staining method was 67.17%, and a higher portion of viable sperm (82.67%) was observed by staining with the Hoechst procedure (P < 0.01). On the basis of the results obtained it is concluded that both methods can be used for detecting the acrosome reaction of frozen-thawed bovine spermatozoa. PMID- 18401965 TI - Detection and analysis of porcine circovirus type 1 in Hungarian wild boars: short communication. AB - Porcine circovirus type 1 (PCV1) is considered to be a non-pathogenic virus detected in cell cultures, vaccines or products used for cell culture preparations, all of them of porcine origin. Serological evidence and genetic studies suggested that PCV1 was widespread in domestic pigs. The presence of PCV1 in wild boars in Germany was also described using serological methods. This paper reports the first detection of PCV1 in Hungarian wild boars. Samples were collected at slaughterhouses and processed for polymerase chain reactions. The complete genome of PCV1 detected in the samples was determined and compared with the available PCV1 sequences of the GenBank database. The genomes formed two distinct clusters with minimum differences, where the Hungarian wild boar PCV1 (WB-H8) grouped together with genomes originating from domestic swine from China and Australia and with a genome detected in a porcine pepsin product. PMID- 18401966 TI - Mosquitocidal and repellent activity of the leaf extract of Citrullus vulgaris (cucurbitaceae) against the malarial vector, Anopheles stephensi liston (diptera culicidae). AB - The efficacy of the Cucurbitaceous plant Citrullus vulgaris against the Anopheles stephensi was assessed in the present study. The leaf extract of Citrullus vulgaris with different solvents viz, benzene, petroleum ether, ethyl acetate and methanol were tested for larvicidial, ovicidal, repellent and insect growth regulatory activities against Anopheles stephensi. The larval mortality was observed after 24 h exposure. The LC50 values are 18.56, 48.51, 49.57 and 50.32 ppm respectively. The mean percent hatchability of the egg of Anopheles stephensi were observed after 48 h. 100 per cent mortality was exerted at 250 ppm with benzene extract and the other extracts exerted 100 percent mortality at 300 ppm. Skin repellent test at 1.0, 2.5 and 5.0 mg per cm2 concentration gave the mean complete protection time ranged from 119.17 to 387.83 minutes with the four different extracts tested. The Citrullus vulgaris plant extract have shown insect growth regulatory activity against Anopheles stephensi at five different test concentrations ranging from 10 to 150 ppm with different solvents and they exhibit the following EI50 values 28.99, 70.02, 106.33 and 84.25 ppm respectively. PMID- 18401967 TI - Myocardial bridging: cases reports. AB - We report 2 cases of Myocardial Bridge (MB). MB, even though a very frequent anomaly (found out in 85% of autopsies), often is asymptomatic. It is hard to diagnose in patients affected by coronary artery disease, seldomly found out on selective coronary angiograms (in 16-20% of chest pain cases). MB can be considered as one of the causes of coronary artery disease. PMID- 18401968 TI - Anatomic relationship between trigeminal nerve and temporomandibular joint. AB - Temporomandibular joint disorders (TMD) is a collective term used to describe pathologic conditions involving temporomandibular joint (TMJ), masticatory muscles and associated structures. Common related complaints include local pain, limited mouth opening and TMJ noises whereas symptoms often associated to TMD with debated pathogenesis enclose earache, headaches, tinnitus and trigeminal like symptoms such as atypical orofacial pain. In particular, TMD trigeminal associated symptoms are intricate, difficult to treat and exert a great impact on everyday life of the patients thus invoking a complex multidisciplinary treatment. In this paper, the authors analyze the anatomic and topographic relationships between the mandibular branch of the trigeminal nerve and the medial aspect of the TMJ capsule in 8 fresh adult cadavers thus resuming a pathologic relationship between atypical trigeminal symptoms and TMD. PMID- 18401969 TI - Pathogenesis, diagnosis, and management of disseminated intravascular coagulation: a literature review. AB - BACKGROUND: Disseminated Intravascular Coagulation (DIC) is an acquired syndrome characterized by systemic intravascular activation of coagulation, leading to deposition of fibrin in the circulation, occurring in the course of severe diseases. OBJECTIVE: To review literature for articles that focus on the pathogenesis, diagnosis, and management of DIC. INFORMATION SOURCES: Selected articles from the Index Medicus data base. STATE OF THE ART: Recent studies have elucidate the pathogenic pathways that can trigger DIC. However, clinical and laboratory diagnosis of the syndrome remains difficult, as there is no single laboratory test, sensitive and specific enough, to allow a definitive diagnosis of DIC. Cornerstone for the treatment of DIC remains the optimal management of the underlying disorder. However, therapeutic interventions based on our present knowledge of the pathogenesis of DIC may be appropriate. CONCLUSION: Recent research on DIC, resulted in the development of diagnostic algorithms for the diagnosis of the syndrome and better supportive therapeutic strategies. PMID- 18401970 TI - Effects of lifestyle modification on adipocytokine levels in obese patients. AB - OBJECTIVE: Adipose tissue is considered an active secretory organ of adipocytokines. The principal aim of our study was to examine the changes in adipocytokines levels after weigth reduction in obese patients. MATERIALS AND METHODS: A population of 126 obese (body mass index > 30) non-diabetic outpatients was analyzed in a prospective way. Before and after three months of a lifestyle modification program, an indirect calorimetry, a tetrapolar electrical bioimpedance, blood pressure, a serial assessment of nutritional intake with 3 days written food records and a series of biochemical analyses were performed. The lifestyle modification program consisted of a daily hypocaloric diet (1520 kcal: 52% of carbohydrates, 25% of lipids and 23% of proteins). The exercise program consisted of an aerobic exercise for at least 3 times per week (60 minutes each). RESULTS: The mean age was 45.6 +/- 16.9 years and the mean BMI 34.5 +/- 5.2, with 33 males (26.2%) and 93 females (73.8%). A total of 88 patients completed the follow up during 3 months, with a percentage of weight loss of 3.1%. Seventy two patients lost weight after treatment (responders) with an average age of 45.4 +/- 16.8 years and a percentage of weight loss of 4.2%. Sixteen patients did not respond (no-responders), with an increase in weight (88.9 +/- 10.5 vs 88.7 +/- 10.9 kg: p < 0.05) and BMI (34.7 +/- 5.4 vs 35.5 +/- 5.5 kg: p < 0.05). The average age (45.9 +/- 15.9 years) of this group was similar than that of responders. In responders (weight loss), BMI, weight, fat mass, glucose, total cholesterol, LDL cholesterol and systolic blood pressure decreased and VO2 increased. After treatment, no statistical differences were detected in energy intake: carbohydrate, fat, and protein. Exercise improved after treatment in responder group. Only serum leptin levels had a significant decrease in responder group (12%). After treatment, no responder group has similar values of all adipokines, including leptin (81.3 +/- 70.6 vs. 76.1 +/- 43 ng/ml). CONCLUSION: Three months of lifestyle modification significantly improved anthropometric and cardiovascular risk factors, regardless of their minimal decrease in energy intake and the weight loss. Additional studies will be need to clarify the contribution of lifestyle modification in changes of serum adipocytokine levels. PMID- 18401971 TI - Active thymopoiesis in idiopathic chronic pancreatitis. AB - BACKGROUND/AIMS: Cellular immunity has a pivotal role in the pathogenesis of chronic pancreatitis (CP), resulting in pancreas infiltration by T-cells. Studies on systemic immunity are few and contradictory. One study reported a decrease of naive CD45RA+ cells. The presence of naive T cells, detected as recent thymic emigrants (RTEs), is evaluated with a new molecular technique by using real-time PCR to detect the T-cell receptor excision circles (TREC). To elucidate the role of naive T-cells in the pathogenesis of CP, we investigated the percentage of sj TREC in CP patients. PATIENTS: Thirty CP patients were studied and compared to 30 sex- and age-matched healthy volunteers. METHODS: Genomic DNA was isolated from peripheral blood mononuclear cells (PBMC) of each patient. RTEs were evaluated by measuring sj-TREC by real-time PCR. RESULTS: The mean percentage of sj-TREC+ cells present in CP was not significantly different from that of control group (0.02319% vs 0.02338%, respectively). CONCLUSION: Our data show that naive TREC+ cells are normally represented in CP. The presence of active thymopoiesis may be the underlying mechanism resulting in continuous production of T-cells, responsible of maintaining the inflammatory process. PMID- 18401972 TI - Anti-phospholipid syndrome: clinical spectrum and therapeutical/prophylactic strategies in the pediatric population. AB - Anti-phospholipid syndrome (APS) is a potentially life-threatening autoimmune condition characterized by the presence of anti-phospholipid antibodies (aPL) giving rise to increased hypercoagulability, which induces venous or arterial thrombotic events at whatever age and recurrent fetal loss in the fertile age. Antigens that are targeted by aPL include cardiolipin and beta2-glycoprotein I. Primary APS is defined in the absence of an underlying disease, while secondary APS is observed in the context of another established pathological condition. APS has a wide variety of clinical signs and serological characteristics. This paper describes the current approaches towards diagnosis, therapeutic modalities and secondary prevention applied to children. PMID- 18401973 TI - Specimen removal after laparoscopic appendectomy: a cheap trick. AB - Laparoscopic appendectomy (LA) has gained worldwide acceptance as an effective surgical treatment of acute appendicitis. One of its main advantages is the almost total absence of cosmetic sequelae. Obviously, this relates with the use of little sized trocars. Whereas the whole dissection can be performed by means of two 5-mm operative ports, other than the 10-mm laparoscope one, usually the dissected appendix cannot be removed through a 5-mm trocar. As in most cases a 5 mm camera is not available, the surgeon must use a second 10-mm trocar, thus losing the cosmetic advantages. In this technical note a cheap trick to retrieve the specimen after LA is described. PMID- 18401975 TI - HCV co-infection does not affect the TREC/IL-7 pathway in HIV disease. AB - About thymic output, little is known in HIV-HCV co-infected patients. Thymic output can be measured by T-cell receptor excision circles (TREC) present in the so called "recent thymic emigrants" (RTEs). We have analyzed, by Real time PCR, sj-TREC+ cells in 11 patients with HIV-HCV co-infection; all patients were treated with highly active antiretroviral therapy (HAART), but were naive for interferon anti-HCV treatment. The results were compared with those of 21 age matched normal donors. These data show no reduction of sj-TREC+ cells in co infection. In 5 co-infected patients, IL-7 plasmatic levels were also evaluated by ELISA and no difference between co-infected patients and normal controls was found. Taken together, our data, although limited by the numerosity of the sample, may suggest that HCV co-infection does not affect the TREC/IL-7 pathway in HIV disease. PMID- 18401976 TI - [The use of PET/CT in the diagnostics of lung cancer]. AB - More than 6000 new cases of lung cancer are diagnosed annually in the Czech Republic. While incidence in men remains stagnant in recent years, incidence in women steadily grows. Combined imaging modality positron-emission tomography (PET) and computed tomography (CT) becomes the imaging method of choice for an increasing number of oncology indications, including lung cancer. With this integrated PET/CT device, accurately registered anatomic and functional images can be acquired in a single examination. Having such aligned image sets available has been shown to increase the accuracy of the interpretation of pulmonary nodules and pulmonary masses. PMID- 18401974 TI - The efficacy and safety of a single dose of polyhexamethylene biguanide gynaecologic solution versus a seven-dose regimen of vaginal clindamycin cream in patients with bacterial vaginosis. AB - OBJECTIVE: At the present the clinical treatment of choice of bacterial vaginosis (BV) is the use of systemic or local metronidazole or clindamycin. Aim of the study was to evaluate the efficacy and tolerability of a single dose of gynaecologic solution, Polyhexamethylene Biguanide (PHMB), Monogin, in the treatment of BV in comparison to a 7-days treatment with clindamycin vaginal cream. STUDY DESIGN: This multicenter, randomized, single-blind, parallel-group study enrolled 740 patients with BV infections. Treatment consisted of either a single intravaginal dose of PHMB or 7 daily doses of Clindamycin. Efficacy and safety were assessed 21-30 days after the start of treatment. The efficacy endpoints were Investigator Cure, Clinical Cure (a composite of all 4 Amsel's criteria and investigator Cure), Nugent Cure (Nugent score < 4), and therapeutic cure (a composite of clinical cure and Nugent Cure). Resolution of individual Amsel's criteria was also evaluated. Any adverse event of the treatment has been monitored throughout the study. RESULTS: No significant differences has been reported in cure rates between the PHMB and Clindamycin treatment groups in Investigator Cure (P = 0.702), Clinical Cure (P = 0.945), Nugent Cure (P = 0.788), or Therapeutic Cure (P = 0.572). Results were also similar for 3 of 4 and 2 of 4 Amsel's criteria and for each individual Amsel's criterion (all P-values > 0.200). Ninety-five percent confidence intervals for each endpoint were consistent with equivalence between the 2 products. There was no significant difference between the treatment groups in the incidence of treatment-emergent adverse events (P = 0.386). CONCLUSIONS: A single dose, of PHMB gynaecologic solution (Monogin) is equivalent in safety and efficacy to a 7-dose regimen of Clindamycin vaginal cream in the treatment of bacterial vaginosis. Futhermore the compliance as been reported to be higher for the single-dose treatment with PHMB than with 7-days treatment with Clindamycin. PMID- 18401977 TI - [The Rubinstein-Taybi syndrome or a broad thumb-hallux syndrome]. AB - The Rubinstein-Taybi syndrome (broad thumb-hallux syndrome) is a rare congenital disease with prevalence 1:125,000 of life-born children. It is characterised by multiplex malformations, which includes growth and psychomotor retardation. Up to this date, over 1000 cases have been described in literature. This review is mainly focused on a description of symptoms, which occur in the syndrome mentioned above and serve as main diagnostic markers. Mutations in the CBP and the p300 genes have been associated with this disease as well. Therefore, substantial part is devoted to aetiology, where emphasis is put on a genetic origin of the Rubinstein-Taybi syndrome. Possibilities of this diagnose are mentioned at the end of the article. PMID- 18401978 TI - [Problems of surrogate decision-making. 1st part--absence of mandatory directive for treatment]. AB - Recently, due to the progress of modern medicine, physicians get into situation when the patient is unable to decide about his treatment, but the decision has to be done and moreover with all respect to fundamental principles of medical ethics. There are two principal questions: "How to make the decision?" and "Who should make the decision". The answer is more complicated and in this article I am trying to show some possible approaches. PMID- 18401979 TI - [Problems of surrogate decision-making. 2nd part--presence of mandatory directive for treatment]. AB - Recently, due to the progress of modern medicine, physicians get into situation when the patient is unable to decide about his treatment, but the decision has to be done and moreover with all respect to fundamental principles of medical ethics. There are two principal questions: "How to make the decision?" and "Who should make the decision". The answer is more complicated and in this article I am trying to show some possible approaches. PMID- 18401980 TI - [Spiritual phenomena occurring in everybody and health]. AB - The past several years have seen an explosion of research in the area of spirituality and health. However, confusion and incomprehension of the conception of spirituality (e.g. confounding spirituality with various conventional views on religiousness) hampers better understanding in this area. The present paper proposes definition of spiritual phenomena in man based on natural epistemological and instrumental criteria (whether a certain phenomenon can be objectively known and evoked): spiritual phenomena in man are those, which cannot be objectively known nor evoked, but which act (e.g., love, idea). Spiritual phenomena can be really known only in the self ("in spirit"). Objectively known can be only manifestations of spiritual phenomena. Some attributes of love (e.g. its personal uniqueness) or ideas (e.g., sense of own life) whose satisfaction appears to be important for health are briefly outlined. A review of some frequently cited recent papers investigating the role of spirituality in health and discussion of frequent pitfalls in this area is given. Spirituality is a universal human phenomenon. All human beings, secular or religious, encounter with spiritual phenomena. Although the present conception of spirituality distances from some conventional views on religiousness, it is not atheistic. On the contrary, it accommodates the basic religious concept "God is love". Conceptual clarification is essential for further progress in the study of impact of spirituality on health. PMID- 18401981 TI - [Tuberculosis among foreigners in the Czech Republic 2003-2006]. AB - Authors compared the situation in tuberculosis among foreigners in the Czech Republic during the years 2003-2006. Foreigners participated in 13.0-14.2% of total notified tuberculosis cases during this period. Proportion of smear positive cases among foreigners in this period grew up from 32.8% to 40.8%. The proportion of active case finding of tuberculosis cases among foreigners declined from 36.0% to 15.4%. At the same time foreigners formed an important part in drug resistance of TB-strains in the Czech Republic. 50% of MDR-TB cases were detected in foreigners. In 20 MDR-TB cases notified in the years 2003-2006 10 were detected in foreigners. If the number of immigrants from countries with high prevalence of tuberculosis to the Czech Republic increases the control of TB in the Czech Republic could be negatively influenced. PMID- 18401982 TI - [Are we threatened by misuse? Discreditation of medical genetics]. PMID- 18401983 TI - [Kawasaki disease and its ocular manifestations]. AB - The article describes one of the occurring types of vasculitis in children. Discussing the essential manifestations of the disease it focuses on ocular symptoms as one of the major criteria. The diagnosis is often done on the basis of a clinical description further specified by laboratory and instrument techniques. Awareness of the symptoms typical of the disease, frequently identifiable even macroscopically, facilitates the early diagnosis. A differential diagnosis of febrile responses may include a wide range of possibilities considered including faulty conclusions consequently causing a delay in therapy, which may be fatal. With the Kawasaki disease a bilateral injection of bulbar conjunctiva is the first signal of objectification followed by frequent acute bilateral iridocyclitis with delayed development and prolonged duration. Thus the problem of a red eye and febrile responses may suggest the above-mentioned serious diagnosis, which is at present, however, well-curable and recognisable with stress put on the time factor. PMID- 18401984 TI - [Corneille Jean Francois Heymans (1892-1968)]. PMID- 18401985 TI - [Regional anaesthesia and cancer recurrence risk]. PMID- 18401986 TI - [Inflammation, cancer--and anesthesia?]. PMID- 18401987 TI - [General anesthesia in mentally disabled patients undergoing dental surgery]. AB - OBJECTIVE: To describe the anesthetic technique used, the evaluation of airway patency, and the perioperative complications in patients with lower than average intelligence (mentally disabled) who are administered general anesthesia for dental surgery. MATERIAL AND METHODS: We carried out a prospective, descriptive, comparative study of mentally disabled ASA 2-3 patients. The patients were distributed in 2 groups: mild to moderate mental disability and severe to very severe mental disability. Induction was via intravenous or inhaled anesthesia depending on availability of venous access. Maintenance was with sevoflurane in oxygen and air at variable concentrations in order to maintain a bispectral index (BIS) between 40 and 60. Statistical comparisons were based on the chi2 test, the log-rank test and the t test. RESULTS: Forty-seven patients were enrolled. The anesthetic technique maintained hemodynamic stability in both groups. No statistically significant differences were found in Mallampati classification, Cormack-Lehane classification, or level of disability. The incidence of complications was higher in the group with more severe disability; the most common complication was difficult tracheal intubation. Bradycardia was the most common complication in the group with mild to moderate mental disability. CONCLUSIONS: The anesthetic technique used in this study proved to be safe and effective in this type of patient. A higher degree of mental disability led to less effective examination of the airway and more difficult direct laryngoscopy. BIS was as effective for monitoring in this population as it is in the general population. PMID- 18401988 TI - [Assessment of agreement between anesthesiologists' location of anatomical landmarks]. AB - OBJECTIVE: The main objective of this study was to evaluate agreement between anesthesiologists' location of specific anatomical landmarks in regional anesthesia. MATERIAL AND METHODS: We administered an anonymous written questionnaire asking for the location of the T5 and T10 dermatomes, Tuffier's line (L3-L4 space), and the T7-T8 and C6-C7 spaces on an anatomical drawing. We asked about each anesthesiologist's number of years of experience and type of block most commonly performed and in what type of surgery, number of blocks performed per year, whether or not levels of sensory and motor blocks were assessed, and whether training had been received. Relative frequencies were then calculated. The relationships between years of experience and number of blocks performed and number of correct identifications of anatomical points were analyzed using the chi2 test. RESULTS: We studied 100 doctors (66 residents and 24 staff anesthesiologists). The landmark for which agreement was highest was T10 (91.8%); agreement was lowest for T5 (38.1%). Forty-five percent of anesthesiologists performed between 100 and 250 neuraxial blocks per year. The most commonly performed procedure was a lumbar block (98% of the anesthesiologists). Level of sensory block was evaluated by 95% of the respondents and level of motor block by 83%. Response to heat and cold was used by 94.8%; the Bromage scale was used by 81%. The number of years of experience and the number of correctly located points appeared to be unrelated (P=.76). Nor was there a correlation between the number of blocks performed and number of correctly located points (P=.2). CONCLUSIONS: T10 was the landmark correctly identified by the largest number of respondents; T5 was correctly identified by the fewest respondents, with answers differing by as much as 4 dermatomes. PMID- 18401989 TI - [Survey of anesthetic techniques used in Catalonia: results of the analysis of 23,136 anesthesias (2003 ANESCAT study)]. AB - OBJECTIVE: To examine anesthesia practice in Catalonia in 2003 and to describe the specific characteristics and distribution of the anesthesia techniques used and the patient profiles. PATIENTS AND METHODS: We used the data from the ANESCAT epidemiological study that gathered information in questionnaire form on all anesthesias performed in Catalonia on 14 randomly selected days representative of practice in 2003. RESULTS: All 131 hospitals authorized to perform anesthesia participated in the study. We collected 23136 questionnaires, from which we extrapolated to estimate 603189 anesthesias for the year and a rate of 9.0 anesthesias per 100 inhabitants per year. Fifty-eight percent of the patients were women and the mean age was 52 years. The physical status of the patients was as follows: ASA 1 or 2, 73.3%; ASA 3, 213%; ASA 4 or 5, 5.4%. The mean duration of anesthesia was 60 minutes. The most common form of anesthesia was a regional block (41.4%) and spinal block was the one performed most often. General anesthesia was used in 33.5% of the cases, combined anesthesia in 3.5%, and sedation in 21.6%. In descending order, orthopedics/trauma, ophthalmology, general surgery, obstetrics, cataract surgery, vaginal delivery, inguinal hernia repair, and colonoscopy were the procedures for which anesthesia was most commonly administered. CONCLUSIONS: Almost 1 in 10 persons in Catalonia are given anesthesia each year and most of the procedures involve locoregional anesthesia or sedation. These data provide a picture of the current situation of anesthesiology and make it possible to forecast future anesthesia requirements. PMID- 18401990 TI - [Maternal satisfaction with the quality of epidural analgesia for pain relief in labor]. AB - OBJECTIVE: To measure patient satisfaction with epidural analgesia in labor and to analyze the sociodemographic characteristics of the obstetric population treated in our hospital. MATERIAL AND METHODS: We administered an anonymous questionnaire in July and December 2003 to all patients who received epidural analgesia during labor to obtain information on the intensity of pain before analgesia and the efficacy of this procedure. RESULTS: A total of 1067 questionnaires were returned. Of the patients who responded to the questionnaire, 91.3% were satisfied with the process of epidural anesthesia, 93.8% stated that they would recommend the technique used in our hospital, and 94% responded that they would request the technique again in our hospital. Spanish nationals accounted for 74.8% of the surveyed patients; the remaining 25.2% were from other countries-mainly from Central and South America (18% of the total). Before administration of epidural analgesia, 23.3% of Spanish primiparas defined labor pain as severe compared to 40.7% of foreign primiparas (P<.001). Of the multiparas, 212% of Spanish patients described the pain as severe compared to 40.4% of foreign women (P<.001). CONCLUSIONS: The overall level of satisfaction with the process of epidural analgesia is very high and was not influenced by sociodemographic factors. Labor pain is perceived as being more intense by non Spanish women. PMID- 18401991 TI - [Anesthesia in malignant hyperthermia]. AB - Malignant hyperthermia is a condition for which the anesthesiologist must be prepared to take specific measures. Associated mortality is high despite correct diagnosis and, occasionally, the right treatment. Malignant hyperthermia is rare in Spain, however, and perhaps for this reason the Spanish public health authorities have not made an effort to respond to persistent calls for a national reference center for study and diagnosis, such as can be found in other European countries. The problem of malignant hyperthermia may become worse in the near future due to changes in the population brought about by immigration. It is therefore necessary to establish appropriately revised protocols and action plans to cope with this possibility. A reference center cannot be created by individuals, but we can become more aware of the need to establish up-to-date protocols in our departments in order to be prepared for this situation. PMID- 18401992 TI - [Perioperative management of a ruptured sinus of Valsalva aneurysm]. AB - Congenital sinus of Valsalva aneurysms are extremely rare in Spain. The lesion consists of a defect that allows the aortic media to separate from the annulus fibrosus of the aortic valve, causing it to dilate with arterial pressure. The natural course of the aneurysm involves the risk of complication due to bacterial endocarditis, with conduction blocks or myocardial ischemia. Rupture of the aneurysm, usually into a right chamber, causes a left-right shunt that leads to heart failure and death if untreated. We present the case of a previously asymptomatic 60-year-old woman who presented with a clinical picture that led to a diagnosis of hyperthyroidism, and in whom there occurred a coincident rupture of a congenital sinus of Valsalva aneurysm. We describe the anesthetic procedure and emphasize the importance of intraoperative echocardiography throughout the resection of the aneurysm. PMID- 18401993 TI - [Anesthetic management in a patient previously diagnosed with takotsubo cardiomyopathy]. AB - Takotsubo cardiomyopathy (transient apical ballooning of the left ventricle) is a recently described and often underdiagnosed entity. The syndrome is observed predominately in postmenopausal women and the clinical signs are similar to those of an acute anterior myocardial infarction. In most of the reported cases an emotional or physical stress event has been identified as a trigger, and perioperative stress has been suggested as the trigger in some of these cases. Outcome is favorable with the right treatment, though recurrences are possible. We report the anesthetic management of a 79-year-old woman with a previously diagnosed episode of Takotsubo cardiomyopathy, who was admitted to our hospital for total hip replacement. Care was taken to provide proper preoperative sedation before provision of hyperbaric spinal anesthesia, followed by sedation with intravenous propofol. Surgery and the early postoperative period were uneventful. We believe that minimizing perioperative anxiety should be a priority in these patients due to the possibility that a catecholamine discharge might trigger an episode of Takotsubo cardiomyopathy. PMID- 18401994 TI - [Sevoflurane and liver dysfunction]. PMID- 18401995 TI - [Pneumothorax secondary to axillary sentinel lymph node biopsy]. PMID- 18401996 TI - [Comment on the letter to the editor "Anesthesia with remifentanil for thoracoscopic thymectomy in a girl with myasthenia gravis"]. PMID- 18401997 TI - [Foreign body aspiration during extubation]. PMID- 18401998 TI - [Ventricular fibrillation in a patient with a type I Brugada syndrome]. PMID- 18401999 TI - [Stents, antiplatelet therapy, and surgery]. PMID- 18402001 TI - [An option for difficult intubation: the Truview EVO2 videolaryngoscope]. PMID- 18402000 TI - [Anesthetic management in a case of recurrent mediastinal paraganglioma]. PMID- 18402002 TI - [Chance finding of a right subclavian artery aneurysm in a preoperative chest film of a patient with dysphagia]. PMID- 18402003 TI - [In situ pinning with Kirschner wires for chronic juvenile slipped capital femoral epiphysis]. AB - Fixation in situis the most common method of management for different degrees of slip of the proximal capital femoral epiphysis. The aim of our study is to retrospectively analyze final results and complications after in situ pinning with Kirschner wires for stabile slipped capital femoral epiphysis. We retrospectively reviewed clinical records and radiographs of 61 children (17 girls and 33 boys) at the average age of 11.8 years. The average follow-up period was 7.4 years, minimum 3 years. In slips 300 and less (38 hips) vast majority of patients had very good and good subjective results (pain and function), 5% and 8% has satisfactory objective (limping and range of motion) and radiological results respectively. In slips between 30-60 degrees (23 hips) 75%, 60% and 70% had satisfactory subjective, objective and radiological results respectively. One child had chodrolysis that resolved at later follow up. There was no case of AVN. In 6 cases wires migration was found. In situ spinning with Kirschnerwires is simple safe and gives satisfactory results. This method of treatment especially recommended in younger, skeletally immature children with remaining growth of femoral neck. PMID- 18402004 TI - [Results of the surgical treatment of ankle fractures in children and adolescents]. AB - We present results of treatment in 36 cases of distal tibial epiphyseolysis in 34 patients treated between 1999 and 2005 with a minimal follow up of 12 months (average 28,5, range 13-84 months). The mean age at injury was 12 years (range from 7 to 16). There were 22 cases of type II injury according to the Salter Harris classification, 5 cases of type VII, 3 of type IV, 2 of type I and two cases of triplane fracture, one case of retrospectively recognised type V and one of type III. 12 children required open reduction and K-wire fixation, the rest of 22 children underwent closed reduction followed by percutaneus fixation in 12 cases. Concomitant fibular fracture occured in 30 cases and required open reduction and fixation in 4. There were 18 good, 14 fair and 3 bad results according to Gleizes. The mean AOFAS scale result was 92.7. One child underwent 2 weeks of intravenous antybiotic therapy to recover from superficial skin infection around percutaneus K-wires. Four cases of physeal bar formation were noted. In two of them distal tibial osteotomy to correct ankle varus was necessary and one underwent bony bridge removal followed by fatty tissue interposition. The remaining physeal bar resulted in 10 degree of ankle valgus without functional dispairement. PMID- 18402006 TI - [The three-dimensional pattern of the pelvis movement in standarized gait cycle in patients before and after knee arthroplasty--leading article]. AB - The main goal of this study was to estimate the influence of knee arthroplasty on angular changes of pelvis movement in three planes. The three-dimensional analysis of locomotion was carried out with 13 patients afer total knee replacement aged 47-74, using Vicon system. The first examination took place before knee operation and the second examination 6 months after arthroplasty. The results were represented at the background of biomechanical gait parameters of 30 healthy people aged 50-70. The angular changes of pelvis movement in each gait phase were taken into consideration. After knee arthroplasty the results of patients examinations were similar to the biomechanical norm in sagittal and transversal plane. In the sagittal plane the difference between the results of the first and the second research was 15 degrees. The first examination of pelvis movement in transversal plane showed that the distinction between patients results and biomechanical norm was 17 degrees in initial contact and terminal swing; after total knee replacement the movement amplitude was similar to the biomechanical norm. The biggest change of the hip joint movement after knee replacement was noticeable in transversal plane but there was only slight improvement in sagittal and frontal plane. There was also noticeable change in the knee movement in frontal plane and the results of the two research sessions in sagittal and transversal plane was similar to the biomechanical norm. PMID- 18402005 TI - [Remote results and failures operating treatment peritrochanteric femur fractures using dynamic hip stabilizer]. AB - The authors make distant evaluation of functional and radiological outcomes after operative treatment of peritrochanteric femur fractures using dynamic hip stabilizer (DSB) from perticulary to take into consideration complications and failures. It was done basing on clinical material of 1998-2005 years. 142 patients were included to the study, 89 women (62.7%) and 53 men (37.3%), aged between 53-97 years (average 77 years). These patients make up 72.5% of all treated because of peritrochanteric femur fractures. Boyd and Gryffin classification was used for evaluation of type of fracture, Harris criterions was used for evaluation of functional outcomes and Golec's criterions was used for evaluation of radiological outcomes. Obtained outcomes allow confirm usefulness of DSB for treatment of type I and II according to Boyd and Griffin classification. Poor outcomes were obtained in type III and IV. PMID- 18402007 TI - [Prognostic value of Lauge-Hansen and Danis-Weber classification in nonoperative and operative tibia ankle fracture treatment]. AB - Tibia ankle fractures and traumatic injuries of the ankle-tibia joint area are one of the most intractable problems in orthopedic and traumatic surgery. Because of the displacement of these fractures they require precise opening of the joint surface area to undertake the early repositioning in order to gain eficient walk. The objective of this paper was to evaluate and compare the final results of nonoperative and operative treament methods, specificlly the two most common prognosis values of tibia ankle fracture clasifications by Danis-Weber and Lauge Hansen. This is the prognositic marking in the range of functional and radiological results dependant upon the method beeing applied. The repetetiveness and accordance of these two classifications has been evaluated as well. This research was conducted on the retrospective evalutaion based on the X-rays of 61 patients (28 women and 33 men) between the ages of 20-76 making the average age 49.5. The definition of accordance and repetitivenes was conducted by three doctors, who evaluated the x-rays from the standard fornt, back, and side projection of relaxed joint ankle. PMID- 18402009 TI - [Effect of intramedullary nailing of tibial shaft fractures on the pressure in the deep posterior compartment of the leg]. AB - In the years of 2003-2005 there was carried out an investigation on 24 patients (17 male and 7 female) based on measuring the pressure in the deep posterior compartment during tibial intramedullary nailing with reaming. The pressure was checked in each part of the operation. The mean increase of the pressure was 7% (+ 1.6 mmHg) and didn't cause any risk of compartment syndrome. In our opinion the intramedullary nailing might be consider as a safe procedure. PMID- 18402008 TI - [Long-term results comparison of simultaneous open reduction and subtalar arthrodesis with the efficacy of closed reduction in the treatment of fresh fractures of calcaneus]. AB - AIM: The aim of the study was the retrospective comparison of results of treatment of fresh fractures of calcaneus with open reduction and simultaneous subtalar arthrodesis performed at Orthopaedic and Traumatology Department in Lublin with closed reduction according to Westhues in Traumatology and Orthopaedic Department in Krosno. MATERIAL AND METHODS: In the years 1994-2001 in Orthopaedics and Traumatology Department in Lublin 33 patients were treated because of 35 fractures of calcaneus (2 bilateral fractures). The comparative material consists of 31 patients treated because of 38 fractures of calcaneus (7 bilateral fractures) in Traumatology and Orthopaedic Department in Krosno in the years 1994-2004. In retrospective study preoperative and postoperative Bohler's angle was measured, as wellas type of fracture was defined according to Essex Lopresti classification. The clinical results of treatmentwas evaluated with Rowe's classification. RESULTS: Long-term evaluation was made to all of the patients treated at Orthopaedic and Traumatology Department in Lublin. The follow up is from 5 to 12 years (average 8 years). Clinical results according to Rowe's classification is from 15 to 100 points (average 84.4 pts.). 17 patients treated in Traumatology and Orthopaedic Department in Krosno with closed reduction according to Westhues were evaluated. The follow up is from 2 to 12 years (average 7 years) with the average 52.35 pts in Rowe's classification. The lack of healing of subtalar arthrodesis was observed in one patient, meanwhile osteoarthritis of subtalar joint was observed at 14 patients after closed reduction according to Westhues. CONCLUSIONS: Closed reduction according to Westhues gives possibility for Bohler's angle restoration, but it does not create the chance for anatomical reduction of articular surface. The considerable percentage of patients suffering pain after Westhues's method testifies about incomplete reduction of articular surface. Open reduction of fracture of calcaneus, combined with simultaneous subtalar arthrodesis restores the correct Bohler's angle and prevents from pain during walking. PMID- 18402010 TI - [Ankle fractures in children and adolescents]. AB - We present a rewiev of orthopaedic literature, our clinical experience, and examples on pediatric ankle fractures. The rules of anatomic physeal and articular surface reduction in intraarticular injuries are emphasised. We present complications, specially ankle axial deformities caused by bony bars and its reasons, specially periosteal interposition or jatrogenic physeal damage by an inapropriate stabilisation. PMID- 18402011 TI - [Prevention of algodystrophy of the upper limb]. AB - Treatment of algodystrophy is a challenge. Early recognition and immediate commencement of effective therapy gives a real chance of recovery, whereas progression into chronic phase is associated with poor prognosis and disability of the limb. Prevention of the condition is a reasonable approach, but little is known about it. Some authors have claimed that careful operative technique, anatomic dissection, avoidance of nerve traction, proper postoperative care and early mobilization of the limb after trauma or operation can reduce risk the frequency of algodystrophy, but this has not been scientifically confirmed. This paper presents current knowledge about prophylaxis of algodystrophy, based on literature and author's own experience. It was stated that, as yet, no specific, effective measures are known to prevent algodystrophy. Administration of 0.5 g vitamin C for 50 days after fracture of the distal radius reduces risk of the condition, but is was reported in only one study. Operation of unstable distal radial fractures, avoiding of tight, uncomfortable plaster casts and painful, forced physiotherapy is believed to reduce risk of algodystrophy, but it is based on observation rather than scientific evidence. Patients who recovered form algodystrophy are not more susceptible to recurrence after further trauma or surgery. An early recognition of incipient algodystrophy and immediate commencement of effective therapy (i.e., with Mannitol and Dexamethasone) was found to be the most important part of prophylaxis of the development of the florid condition. PMID- 18402013 TI - [What is the future of nephrology for nephrologists?]. PMID- 18402012 TI - Is conservative treatment really effective for Legg-Calve-Perthes disease? A critical review of the literature. AB - INTRODUCTION: The treatment of Legg-Calve-Perthes Disease (LCPD) is controversial and not well defined. This literature review aimed to assess the quality of the evidence available to support the effectiveness of conservative LCPD treatment advocated by orthopaedic surgeons and(or) paediatricians. A secondary aim was to see if conservative treatments really modify the natural history of LCPD. MATERIALS AND METHODS: The review was performed mainly on the PubMed Database and based on the principles of Evidence Based Practice (EBP). Keywords used were Perthes disease, conservative treatment, containment treatment, review, and weight-relief. EBP is the integration of the best research evidence with clinical expertise and patient values of which there are five different levels of evidence: (1) Randomized Controlled Trials; (2) Prospective Cohort Study; (3) Case Control or Retrospective Cohort Study;(4) Case Series; 5) Expert Opinion or Individual Case Report. Results. Until 20th August 2005 there were 144 articles of clinical relevance about conservative treatment of LCPD: 16.7% of EBP level 5; 50.7% of level 4; 31.9% of level 3; none of level 2; and 0.7% of level 1. CONCLUSION: The quality of evidence that supports conservative treatment for children with LCPD is not of high quality. There is no scientific evidence that conservative treatments modify LCPD natural history. Containment, no containment and simple symptomatic treatment have comparable effectiveness. Prolonged weight relief and(or) containment treatments are associated with social and psychological problems. PMID- 18402014 TI - [Acute renal failure in 2008]. AB - Prevalence of acute renal failure is rising on account of a aging population. Despite improvements in treatment of renal failure-related complications and renal replacement therapy, its mortality remains elevated. Therefore, earlier detection and prevention of frequent causes of acute renal failure in patients at risk are of paramount importance. PMID- 18402015 TI - [Nephrogenic systemic fibrosis: another problem for patients with chronic renal failure]. AB - Nephrogenic systemic fibrosis, another problem for patients with chronic renal failure Nephrogenic systemic fibrosis is a rare but severe disease described in patients with kidney failure. High morbidity ant mortality are associated with this new condition. Epidemiological studies strongly suggest a link between nephrogenic systemic fibrosis and gadolinium administration for magnetic resonance imaging enhancement. The disease is primarily cutaneous, with oedema affecting the limbs, later evolving to fibrosis that leads to joints contractures. The lesions can spread to the trunk and involve systemic organs like the heart, lungs and muscles. Given the lack of proved efficient therapy, careful evaluation of the risks and benefits of gadolinium administration should be done in patients with kidney disease. If really needed, a highly stable contrast media should be used. PMID- 18402016 TI - [Plasmapheresis: technique, complications and indications]. AB - Plasmapheresis is an extracorporeal technique used to remove pathogenic macromolecules from the plasma. Plasmapheresis is used to treat neurological, renal, hematological as well as systemic diseases, which explains why many different specialties in medicine can be involved. Plasmapheresis has evolved in forty years into a frequently used, relatively safe procedure. Nowadays a large spectrum of different techniques exists, each with its own possible complications. In this article we will give an overview of these different techniques, their complications and indications, in order to familiarize the reader with this fascinating treatment. PMID- 18402017 TI - [Treatment of secondary hyperparathyroidism in renal insufficiency: role of calcitriol, sevelamer and cinacalcet]. AB - Along with the decrease in kidney function arises a secondary hyperparathyroidism, which constitutes one of the most important risk factor for mortality in patients suffering from renal insufficiency. Treating secondary hyperparathyroidism is challenging, as most of the parameters of mineral metabolism are interconnected. We review here the pathophysiology and treatment options of this entity. PMID- 18402018 TI - [Medical care of renal transplant recipients after the first year post transplantation]. AB - Kidney transplant recipients are a growing population in ambulatory care. Medical follow up after the first post transplant year requires a tight collaboration between transplant centers, primary care physicians and community nephrologists. Although kidney transplantation is the treatment of choice for patients in end stage renal failure, no major improvement has been seen in long-term patient and graft survivals. Mortality of kidney transplant recipients remains higher than that of the general population, due to the high incidence of cardiovascular disease, infection and malignancies related to progressive renal failure and also to immunosuppressive treatment. We review here the optimal ambulatory medical care needed by these patients after the first post transplant year. PMID- 18402019 TI - [Practical lessons from group treatment in low back pain patients]. AB - Multidisciplinary group therapies for low back pain patients have been devised for more than 30 years. Various models have been developed during this period, resulting in multiple programmes with varying effectiveness. Taking the opportunity of the establishment of a new programme at the Geneva University Hospitals, we reviewed these various models and listed the most prevailing elements in the literature. The summary presented here illustrates the hurdles in the treatment of low back pain patients. It underlines the numerous orientations available to the clinician and offers some indications for improving the treatment in these patients. PMID- 18402020 TI - [Subungual exostosis]. PMID- 18402021 TI - [Why is the scope of abuse underestimated and the patients insufficiently treated?]. PMID- 18402022 TI - [Breast cancer: recommendations for prevention (2)]. PMID- 18402023 TI - [Response to James Watson on the question of race]. PMID- 18402024 TI - [Analysis of human hair and its relation to geography]. PMID- 18402025 TI - [What I think of managed care (8)]. PMID- 18402026 TI - [The new constitutional article vote in June]. PMID- 18402027 TI - Jump starting your nursing career: toolbox for success. AB - While there is no single path to career advancement in nursing, there are developmental tools and organizational systems to promote nurses' career advancement and to assist in the organization of professional accomplishments. As nurses achieve additional credentialing, certification, licensure and academic credentials, it is important that they list these credentials after their names to market their career accomplishments and expertise. Career tool boxes such as a file system, resume and professional portfolio, are key organizational systems for nurses to keep track of their success and to promote their careers in nursing. PMID- 18402028 TI - Political leadership: politics 101. PMID- 18402029 TI - Leadership architecture: a multi-dimensional approach to succession planning. PMID- 18402031 TI - Decreasing asthmagens in hospitals. PMID- 18402030 TI - Solving the leadership dilemma: where will nursing's next leaders come from? PMID- 18402032 TI - Leading leaders. PMID- 18402033 TI - Learning leadership through the Student Nurses Association. PMID- 18402034 TI - [Treatment tactics of patients with choledocholithiasis complicated with acute cholangitis]. AB - The results of observation and treatment of 137 patients with choledocholithiasis complicated acute cholangitis are presented. The estimated scale APACHE II is adopted for the state severity of patients with choledocholithiasis and cholangitis sigh determination are examined. With multivariant correlative regression analysis the most informative factors are determined, its grade estimation are carrying out. Optimal treatment tactics depended from state severity is definited. It permits to improve the treatment results owing to decrease of postoperative complications frequency, duration of patients therapy in hospital. PMID- 18402035 TI - [Noncomplicated varicose disease: morphological peculiarities, the ways of reflux spread]. AB - The studying results of anatomical variants of the leg large subcutaneous vein (LLSV) and its tributaries structure are adduced, the possible ways of the blood reflux spread are determined. The authors present anatomical classification of varicose disease of the LLSV basin, the epidemiology data of the disease forms incidence are adduced. PMID- 18402036 TI - [Hepatic functional state in live relative donors of its right lobe in perioperative period]. AB - Analysis of hepatic functional state in live relative donors of its right lobe in perioperative period was conducted. Temporary hepatic dysfunction was revealed, manifesting by tolerance to glucose lowering, hepatic proteinsynthetic and excretal functions disorder and cytolytic syndrome. In 2 weeks after the operation these disorders are become compensated but not all indices return to initial level. PMID- 18402037 TI - [Phenotype of haptoglobin and characteristics of biopolymer metabolism in the connective tissue in adhesive peritoneal disease]. AB - Peculiarities of the connective tissue biopolymers metabolism in adhesive peritoneal disease were studied in patients with different phenotype of gaptoglobin. It was established, that the polymers metabolism in the patients does not differ in gaptoglobin phenotype 2-1 and 2-2. The connective tissue biopolymers metabolism disorders in adhesive peritoneal disease is more expressed in patients with gaptoglobin phenotype 1-1 than in those with gaptoglobin phenotype 2-1 and 2-2. Inclination to adhesions formation in gaptoglobin phenotype 1-1 is caused by the connective tissue biopolymers metabolism disorder. PMID- 18402038 TI - [Dynamics of cellular populations in reparation for the burn wounds in patients with subfascial burns and the opening of the shin bones]. AB - The dynamics of reparation of deep burn wounds after performance of ostheonecrectomy, ostheotrepanation and ostheotrepa-nation in conjunction with application of allofibroblasts culture was studied, basing on the morphometric investigations data. The authors have had concluded, that application of ostheotrepanation, solely or in combination with the allofibroblasts culture usage, make the reparation process less durable as well as the term before effective autodermoplasty is possible and stationary treatment period in injured persons, suffering subfascial burns and the shin bones opening. PMID- 18402039 TI - [Pathogenesis of the lethal postoperative cerebral inflammatory-purulent complications in neurooncological patients]. AB - The main and immediate causes of death, pathogenetic mechanisms of postoperative cerebral inflammatory-purulent complications (CIPC) in patients, suffering the brain tumour, were analyzed. The main pathogenetic patterns of the postoperative period course in the CIPC occurrence were studied, basing on the data of analysis of 30 patients, who died after the operation. There was proved, that the sepsis occurrence on the background of postoperative meningoencephalitis means the lethal ending of the disease. There was established, that in occurrence of septic complications, connected with postoperative meningoencephalitis, the most frequently the affection of respiratory and urinary systems occurs. PMID- 18402040 TI - Adaptive Fuzzy Association Rule mining for effective decision support in biomedical applications. AB - Due to complexity of biomedical classification problems, it is impossible to build a perfect classifier with 100% prediction accuracy. Hence a more realistic target is to build an effective Decision Support System (DSS). Here 'effective' means that a DSS should not only predict unseen samples accurately, but also work in a human-understandable way. In this paper, we propose a novel adaptive Fuzzy Association Rules (FARs) mining algorithm, named FARM-DS, to build such a DSS for binary classification problems in the biomedical domain. In the training phase, four steps are executed to mine FARs, which are thereafter used to predict unseen samples in the testing phase. The new FARM-DS algorithm is evaluated on two publicly available medical datasets. The experimental results show that FARM-DS is competitive in terms of prediction accuracy. More importantly, the mined FARs provide strong decision support on disease diagnoses due to their easy interpretability. PMID- 18402041 TI - Bi-level clustering of mixed categorical and numerical biomedical data. AB - Biomedical data sets often have mixed categorical and numerical types, where the former represent semantic information on the objects and the latter represent experimental results. We present the BILCOM algorithm for 'Bi-Level Clustering of Mixed categorical and numerical data types'. BILCOM performs a pseudo-Bayesian process, where the prior is categorical clustering. BILCOM partitions biomedical data sets of mixed types, such as hepatitis, thyroid disease and yeast gene expression data with Gene Ontology annotations, more accurately than if using one type alone. PMID- 18402042 TI - Kernel design for RNA classification using Support Vector Machines. AB - Support Vector Machines (SVMs) are a state-of-the-art machine learning tool widely used in speech recognition, image processing and biological sequence analysis. An essential step in SVMs is to devise a kernel function to compute the similarity between two data points. In this paper we review recent advances of using SVMs for RNA classification. In particular we present a new kernel that takes advantage of both global and local structural information in RNAs and uses the information together to classify RNAs. Experimental results demonstrate the good performance of the new kernel and show that it outperforms existing kernels when applied to classifying non-coding RNA sequences. PMID- 18402043 TI - State-space approach with the maximum likelihood principle to identify the system generating time-course gene expression data of yeast. AB - We use linear Gaussian state-space models to analyse time-course gene expression data of yeast. They are modelled to be generated from hidden state variables in a system. To identify the system, we estimate parameters of the model by EM algorithm and determine the dimension of the state variable by BIC. PMID- 18402044 TI - Text analysis of MEDLINE for discovering functional relationships among genes: evaluation of keyword extraction weighting schemes. AB - One of the key challenges of microarray studies is to derive biological insights from the gene-expression patterns. Clustering genes by functional keyword association can provide direct information about the functional links among genes. However, the quality of the keyword lists significantly affects the clustering results. We compared two keyword weighting schemes: normalised z-score and term frequency-inverse document frequency (TFIDF). Two gene sets were tested to evaluate the effectiveness of the weighting schemes for keyword extraction for gene clustering. Using established measures of cluster quality, the results produced from TFIDF-weighted keywords outperformed those produced from normalised z-score weighted keywords. The optimised algorithms should be useful for partitioning genes from microarray lists into functionally discrete clusters. PMID- 18402045 TI - Simulating the cellular passive transport of glucose using a time-dependent extension of Gillespie algorithm for stochastic pi-calculus. AB - Realistic simulations of the biological systems evolution require a mathematical model of the stochasticity of the involved processes and a formalism for specifying the concurrent nature of the biochemical interactions. A time dependent extension of the Gillespie algorithm implementing the race condition of the stochastic pi-calculus formalism satisfies both these requirements. This paper formulates those modifications to the original Gillespie algorithm necessary when the time dependence of the reaction propensity is due to changes either of volume or temperature. This re-formulation has been incorporated in the framework of stochastic pi-calculus and has been applied to simulate the passive glucose cellular transport. PMID- 18402046 TI - Transductive learning with EM algorithm to classify proteins based on phylogenetic profiles. AB - We proposed a novel method for protein classification based on phylogenetic profiles. Each protein's profile was extended with extra bits encoding the phylogenetic tree structure and the likelihood, in the form of weights on profile indices, of the protein's functional family membership in each of the reference genomes. The extended profiles were then integrated as part of a kernel of a support vector machine, which was trained in a transductive learning scheme using the EM algorithm to update the weights. Classification accuracy was greatly increased when tested on the proteome of Saccharomyces cerevisiae using the MIPS classification as a benchmark. PMID- 18402047 TI - A constraint logic programming approach to associate 1D and 3D structural components for large protein complexes. AB - The paper describes a novel framework, constructed using Constraint Logic Programming (CLP) and parallelism, to determine the association between parts of the primary sequence of a protein and alpha-helices extracted from 3D low resolution descriptions of large protein complexes. The association is determined by extracting constraints from the 3D information, regarding length, relative position and connectivity of helices, and solving these constraints with the guidance of a secondary structure prediction algorithm. Parallelism is employed to enhance performance on large proteins. The framework provides a fast, inexpensive alternative to determine the exact tertiary structure of unknown proteins. PMID- 18402048 TI - A Merge-Decoupling Dead End Elimination algorithm for protein side-chain conformation. AB - Dead End Elimination (DEE) is a technique for eliminating rotamers that can not exist in any global minimum energy configuration for the protein side chain conformation problem. A popular method is Simple Goldstein DEE (SG-DEE) which is fast and eliminates rotamers by considering single residues for possible elimination. We present a Merge-Decoupling DEE (MD-DEE) that further reduces the number of rotamers after SG-DEE. MD-DEE works by forming residue-pairs but is fast and, like SG-DEE, is practical even for large proteins. Our experiments show that MD-DEE achieves further reduction in residue elimination (up to 25%) after SG-DEE. PMID- 18402049 TI - Biomedical text summarisation using concept chains. AB - BioChainSumm is a biomedical text summariser utilising concept chaining (called BioChain) to link semantically-related concepts within biomedical text together. The BioChain process is adapted from existing lexical chaining approaches which chain semantically-related terms rather than concepts. The BioChain concept chains are used to identify salient candidate sentences which are extracted to produce a summary of the biomedical text. The Unified Medical Language System Metathesaurus and Semantic Network semantic resources identify related biomedical concepts. BioChainSumm is evaluated using the ROUGE system along with several existing, publicly-available summarisers. Our results show BioChain provides a promising methodology for biomedical text summarisation. PMID- 18402050 TI - [Fundamental evaluation of HCV core antigen method comparison with Cobas Amplicor HCV monitor v2.0 (high range method)]. AB - Quantitative measurement of hepatitis C virus (HCV) has been performed by PCR method. However, PCR method has problems such as a special instrument, a complicated manual skill and a high cost. Recently, simple and highly sensitive HCV core antigen (Ag) method has been developed. We performed fundamental evaluation of HCV core Ag method, and compared HCV core Ag method with HCV PCR high-range method. The intra-assay and inter-assay variation coefficients for HCV core Ag were calculated to be within the ranges of 1.0-11.3% and 0.8-9.3%, respectively. The test of dilution linearity revealed the unstableness in the vicinity of a cut-off level of 50 fmol/L. Based on the result of the high-range method; sensitivity, specificity, positive predictive value, negative predictive value, and agreement rate were 97.0%, 100%, 100%, 82.0%, and 96.5%, respectively. The correlation between the HCV core Ag method and the high-range method was r = 0.87. Cost per sample and time from sample preparation to final report for HCV core Ag were cheaper and shorter than those of HCV PCR method, respectively. We consider that the HCV core Ag method seems to be useful as the quantitative measurement of HCV with respect to rapidness, easiness and low cost. PMID- 18402051 TI - [Pathogenesis and therapy for transthyretin related amyloidosis]. AB - Transthyretin (TTR) is a beta-sheet rich protein whose plasma half life is 1.9 days. It behaves as a tetramer and binds to retinol binding protein (RBP) and thyroxin in plasma. Since TTR is a tryptophan-rich-protein, the protein is used as a useful marker protein for nutrition supporting team (NST). However, TTR is also an anti-acute phase protein, and its concentration is influenced by various conditions, such as inflammation and infection. Mutated forms of TTR are the precursor protein of familial amyloidotic polyneuropathy (FAP). Since plasma TTR is predominantly synthesized by the liver, liver transplantation has been performed as an effective therapy for FAP. However, the surgery has several problems, so we must develop novel essential therapies for FAP. Single stranded oligonucleotides (SSOs) or short interference RNA (SiRNA) is a promising option for an essential therapy for FAP. In mutated TTR, instability of tetrameric form of TTR occurs, resulting in misfolding of TTR molecule, which lead to amyloid fibril formation. Since mutated TTR exposes criptic epitopes on the surface of TTR molecule, induction of an antibody for the epitopes was thought to be effective. We synthesized ATTR Y78P, a spontaneously misfolded TTR, and injected it to amyloid laden transgenic mice having human ATTR V30M to induce the antibody for amyloid fibrils. As we expected, amyloid deposition was significantly reduced by the injection of ATTR Y78P to the mice. These therapies may become novel strategies for essential FAP therapy instead of liver transplantation. PMID- 18402052 TI - [Implication of neuron-glia interaction in the molecular mechanism of drug dependence]. PMID- 18402053 TI - [Electrophysiological technique--significance as an experimental method for alcoholic study]. PMID- 18402054 TI - [Difference in medical history classified by ICD-10 between male and female alcoholics]. AB - The drinking history and current medical history of patients with alcohol dependence were surveyed and they were analyzed by gender, age and changes with time (2 stages). The results showed that in the course of continued habitual drinking by patients with alcohol dependence, a wide range of physical complications occurred. The main complications in men were gastrointestinal diseases and in women were mental and behavioral disorders, showing a gender difference in the medical history. This result suggested that there is a high possibility that this will contribute to early discovery and early measures against alcohol related problems in women, which are difficult to bring out into the open. Better alcohol education including mental health is important from an early age. PMID- 18402055 TI - Ethanol-induced JNK activation suppressed via active Akt in hepatocytes. AB - Ethanol induces c-Jun N-terminal kinase (JNK) activation leading to cell death in hepatocytes. However, acute alcohol exposure does not induce remarked cell death in hepatocytes. We hypothesized that active Akt may suppress JNK activation. To clarify this point, we evaluated the role of active Akt in JNK activation under treatment with hepatocyte growth factor (HGF) and compared it with ethanol treatment. Primary rat hepatocytes were treated with 10 ng/ml HGF. 10 min after that, 5 microM insulin, an activator of the Akt pathway, and/or 5 microM LY294002, an inhibitor of the pathway, were added. Hepatocytes were treated with 100 mM ethanol and LY294002. HGF treatment increased JNK activities in hepatocytes. This JNK activation was accumulated by addition of LY294002. These finding suggest that active Akt suppresses JNK activation induced by HGF. On the other hand, addition of insulin did not decrease the JNK activity, showing that insulin-induced Akt activation may rather increase JNK activity. Ethanol also induced JNK activation and this JNK activation was enhanced by LY294002 similar to HGF treatment. We found that active Akt suppressed JNK activation induced by ethanol as well as HGF in hepatocytes. JNK activation may be suppressed by prolonged active Akt or basal active Akt, rather than peaked activation of Akt induced by insulin stimulation. Our results suggest that the suppression of JNK by active Akt may prevent cell death in acute alcohol intoxication. PMID- 18402056 TI - [Is adolescent tobacco use a gateway drug to adult alcohol abuse? A Japanese longitudinal prospective study on adolescent drinking]. AB - This study is the first prospective study correlating smoking and drinking among Japanese adolescents. The aim of the study is to determine if adolescent smoking experience predicts future adult alcohol abuse. We conducted a longitudinal prospective study on adolescent drinking from 1997 to 2007. The study subjects were about 800 male and female junior high school students. Their mean age was 13.5 years old at the starting point. We conducted surveys by post on the subjects concerning drinking status, alcohol-related problems and smoking experience once a year for 10 years. This study attempted to determine if smoking experiences in 1999 (mean age, 15.8 years old) correlated with alcohol abuse in 2005 (mean age, 22 years old) among the subjects of the prospective study. The result showed that male students who had had smoking experiences in 1999 were more likely to become alcohol abusers in 2005 than those without smoking experiences according to the AUDIT. On the other hand, females with smoking experiences as teenagers showed no correlation with alcohol abuse as young adults. Adolescent tobacco use could be called a gateway drug to adult alcohol abuse among Japanese males, but not among females. PMID- 18402057 TI - [Self-disclosure on the Internet: the effects of anonymity of the self and the other]. AB - The effects of anonymity on self-disclosure were investigated in a CMC (computer mediated communication) situation by separately manipulating the anonymity of the self and the other. It was hypothesized that anonymity of the self would enhance disclosure, whereas anonymity of the other would decrease it. Female undergraduate students (n = 60) were randomly assigned to a 2 (self: anonymous or non-anonymous) x 2 (other: anonymous or non-anonymous) experimental design. The anonymity of the self and the other was manipulated in the profile, such as the photo, name, and sex of the self or the other, on the computer screen. The results indicated that anonymity of the self decreased the feeling of anxiety, but had no effect on disclosure, whereas anonymity of the other decreased the feeling of closeness and decreased the intimacy of disclosure. PMID- 18402058 TI - [Facial expressions in the course of relationship formation]. AB - Previous studies investigated the effects of interpersonal relationships on facial expressions by comparing friends with strangers. This study examined facial expressions in the course of relationship formation. Twenty pairs of female strangers met once a week for three weeks watching film clips aimed to elicit a positive emotions. We measured smiles and gazes during and after presentation of the film clips. Subjective emotions and social motives were also measured. Smiling increased from Week one to Week two, but did not change from Week two to three. The results support the theory of Berg & Clark's (1986) that relationships are differentiated very early on. The score for "concern about partner's evaluation" on the social motive questionnaire decreased from Week one to two suggesting that decreasing avoidant social motives facilitates the expressions of smiles in the course of relationship formation. PMID- 18402059 TI - [The structure of interaction in romantic relationships: hierarchical data analysis of inter-subjectivity between partners]. AB - A hierarchical data analysis was conducted using data from couples to examine how self-reports of interactions between partners in romantic relationships predict the quality of the relationships. Whereas the social exchange theory has elucidated the quality of relationships from the individual level of subjectivity, this study focused on the structure of interactions between the partners (i.e., the frequency, strength, and diversity) through a process of inter-subjectivity at the couple level. A multilevel covariance structure analysis of 194 university students involved in romantic relationships revealed that the quality of relationships was mainly related to the strength and the diversity of interactions at the couple level, rather than the strength of interactions at the individual level. These results indicate that the inter subjective process in romantic relationships may primarily explain the quality of relationships. PMID- 18402060 TI - [Contextual cueing in repetition of global or local layouts]. AB - Visual context, such as an association between a target location and a distractor configuration, has been shown to be implicitly learned through repeated experiences in a visual search task, and facilitates the search performance (contextual cueing effect). In the present study, we examined whether the whole layout or only the local half-layout contributed to contextual cueing. In a training session, the whole layout or the local half-layout including the target location was repeated. Then in a test session, half of the layout including the target location was presented while the remaining half of the layout was randomly changed. The results showed that the search performance in the repeated whole layout condition was equivalent to the condition where only half of the layout was repeated throughout both the training and the test sessions. This suggests that the local layout around the target location was selectively learned and used for contextual cueing even if the whole layout was repeated. PMID- 18402061 TI - [Development of semantic knowledge in children's associative false memory]. AB - In the Deese-Roediger-McDermott (DRM) procedure, false recall of a word that was not presented (the critical-lure) can be produced when participants study a list of associative words related to the critical-lure. Recently, some studies using the DRM procedure showed that young children did not produce false recall. The present study hypothesized that the children did not produce false recall in those studies because the lists of words did not reflect the children's associative knowledge. To test this possibility, the present study developed lists that reflect the associative knowledge of five-year-old children and examined false recall using the DRM procedure. The results showed that children falsely recalled the critical-lure after studying the lists that reflected the children's associative knowledge, while they did not recall the critical-lure after studying the lists that reflected adults' associative knowledge. The results indicate that children produce false recall when the lists of words reflect those children's associative knowledge. The present finding suggests that the structuring of semantic knowledge that mediates false recall of the critical lure has developed five years of age. PMID- 18402062 TI - [Visual-haptic simultaneity judgments for dynamic deformation]. AB - Simultaneity is important in cross-modal information processing. However, it is still unclear how simultaneity is perceived between different sensory modalities. Various factors such as spatial location or attention are known to affect simultaneity judgments. In the present study, we focused on the simultaneity judgments of dynamic events, and investigated what kinds of dynamic properties affect these judgments. We presented the deformation of a virtual object in vision and haptics with various stimulus onset asynchronies. Participants judged whether the deformation occurred simultaneously. We measured the effects of duration, velocity, and the amount of deformation on the visual-haptic simultaneity judgments. The results showed that the point of subjective simultaneity changed depending on the duration of deformation. For a shorter duration (400 ms), the visual deformation needed to precede the haptic one to be judged as simultaneous, while for a longer duration (800 ms, 1 200 ms), the asymmetry was diminished, suggesting that information relevant to the duration of the event was used for the vision-haptics simultaneity judgments of dynamic events. PMID- 18402063 TI - [Coping for paranoid ideation in college students]. AB - Research has shown that individual with a high level of paranoia use more avoidant coping strategies (Ellett, Lopes, & Chadwick, 2003). The present study examines coping strategies in 141 college students with lower or higher levels of paranoid ideation using the Paranoia Scale (Fenigstein & Vanable, 1992) and the Interpersonal Stress Coping Inventory (ISI; Kato, 2000). Participants were first required to complete the ISI for paranoid ideation (ISI-pre). They were then asked to complete ISI again (ISI-post) on the assumption that the previous coping strategies (ISI-pre) did not work well. The results showed that people with a higher level of paranoid ideation reported more negative relationship-oriented strategies on both the ISI-pre and ISI-post. The ISI-pre was higher than ISI post for positive relationship-oriented strategies among the participants with a higher level of paranoid ideation. These results suggested that people with paranoid ideation keep using the negative relationship-oriented strategies and reduce positive relationship-oriented strategies, if the primary coping strategies do not work well. PMID- 18402064 TI - [The influences of laterality on global precedence: interference effects]. AB - This study aimed to clarify the influence of laterality on interference effects for global precedence, and to examine the mechanism of these effects. In most previous studies, the neutral condition was excluded from the consistency factor and the difference of latencies between the consistent and inconsistent conditions was regarded as the interference score, which may not be accurate. In this study, the difference of latencies between the neutral and the inconsistent condition was regarded as the interference score. The results of the analysis of variance (ANOVA) for latencies showed that interference effects influenced only right visual field-left hemisphere (RVF-LH), and facilitation effects influenced only left visual field-right hemisphere (LVF-RH) in the local condition. These findings indicate that it does not matter how optimal processing influenced interference effects on suboptimal processing in each hemisphere, for example how global processing influenced local processing in the right hemisphere, but rather how each hemisphere controlled optimal processing. PMID- 18402065 TI - [Young-onset dementia: an unresolved challenge]. AB - Younger people with dementia present a unique challenge to modern Japanese society and those individuals who care for them. Although illnesses causing dementia occur much less commonly in younger than older people, it was estimated that there were about 30,000 affected younger people in 1995 in Japan. For younger people with dementia, effects on families, the presence of dependent young children, and the economic implications are particular challenges. In this article, firstly, epidemiological findings regarding illnesses causing dementia in younger people were described. Secondly, the three major degenerative dementia forms that often develop in presenescence were reviewed. Thirdly, the issue of service planning for such cases was discussed. PMID- 18402066 TI - [Practical use of a rating scale for hospitalization in psychiatric services--the inter-rater reliability of the Japanese version of level of care utilization system for psychiatric and addiction services adult version 2000 (LOCUS)]. PMID- 18402067 TI - [Idiopathic normal pressure hydrocephalus--overviews and pathogenesis]. AB - Idiopathic normal pressure hydrocephalus (iNPH) is a syndrome of gait disturbance, dementia and urinary incontinence without any preceding disorders in elderly patients. Rapid increase of elderly population in Japan prompted us to make guidelines for diagnosis and management of iNPH. The Japanese guidelines were published in 2004 and their characteristics were summarized as follows; 1) Simple and treatment-oriented, 2) Three diagnostic levels: possible, probable and (definite); Shunt surgery is indicated for probable iNPH only, 3) Clinical significance of gait disturbance as the most frequent and treatable symptom, 4) CSF tap test as a keystep of diagnosis, 5) Recommend the use of a programmable valve for prevention of overdrainage, and 6) Flow-charts for diagnosis, shunt indication and prevention of shunt complication. The International guidelines of iNPH were published in 2005. They were created based on evidence-based medicine, but there are some differences. Major differences were in the concept of iNPH, age, range of CSF pressure, MRI findings, and evaluation of CSF tap test, outflow resistance measurement and continuous CSF drainage. This difference would be derived from the weight of the accuracy, popularity and invasiveness of diagnostic tests. At present the pathogenesis of iNPH is not known. A new proposal in which CSF is mainly absorbed in the capillaries in all of the central nervous system may contribute to clarify the pathogenesis in iNPH. PMID- 18402068 TI - [Gait disturbance in idiopathic normal pressure hydrocephalus]. AB - The triad symptoms of idiopathic normal pressure hydrocephalus (iNPH), i.e., dementia, gait disturbance, and urinary incontinence may appear commonly in the elderly suffering from various brain diseases as well as in those with musculoskeletal disorders. It would not be easy to differentiate iNPH from other neurological diseases with musculoskeletal disorders in the elderly. Gait disturbance is an early and central clinical symptom, and its improvement after temporary removal of CSF and shunt surgery is the most notable. This paper reviews the nature, differential diagnosis, pathophysiology, and evaluation of iNPH gait. Broad-based, short-step, magnetic gait with start hesitation and increased instability on turning, which is often expressed as apraxic/ataxic gait, is the cardinal sign of iNPH. Although iNPH gait shares the features with gait in Parkinson disease, progressive supranuclear palsy, and cerebellar ataxia, the nature of iNPH gait is the closest to apraxia of gait, which is characterized primarily by gait ignition failure. The semiological similarity to apraxia of gait and the close relationship to frontal symptoms indicate that the iNPH gait is probably caused by frontal dysfunction. The gait subscale of the idiopathic normal pressure hydrocephalus grading scale and the timed "Up and Go" test are recommended as useful tools for measuring gait disturbance in iNPH. PMID- 18402069 TI - [Cognitive impairment in patients with idiopathic normal pressure hydrocephalus]. AB - The major cognitive impairment in patients with idiopathic normal pressure hydrocephalus (iNPH) is frontal lobe symptoms, such as psychomotor slowing and impairment of attention, working memory, verbal fluency and executive function. Compared with Alzheimer's disease (AD), frontal lobe symptoms are disproportionately severe and the impairment of memory and orientation is disproportionately mild in patients with iNPH. Concerning memory impairment, recognition memory is relatively preserved compared with recall in iNPH. The cognitive impairment of iNPH could be classified into subcortical dementia type. However, compared with Binswanger's disease, impairment of memory and visuospatial attention in NPH may be more severe. Agraphia is also found in some iNPH patients. Thus, iNPH patients could have not only cognitive impairment of subcortical but also cortical types. After shunt operation, cognitive impairment improves but marked improvement in cognitive functions is less frequent than that of the gait disorder. The impairment of memory, working memory and visuoconstructive and psychomotor slowing appears likely to respond shunt surgery. However, in iNPH patients with impairment of memory and either of visuoconstructive or executive function, improvement of overall cognitive deficits after shunt surgery is less, suggesting that a higher degree of neural impairment may identify less reversible injury or that superimposed AD might limit the potential to show improvement. Neuroanatomical bases of cognitive impairment of iNPH are not well documented. However, damage of periventricular structures, such as corpus callosum, frontal subcortical and cortical areas, thalamus, basal ganglia and hippocampus, could be associated with cognitive impairment of iNPH. PMID- 18402070 TI - [Urinary dysfunction in idiopathic normal pressure hydrocephalus]. AB - Urinary incontinence (UI) is a significant burden in the elderly and their caregivers when assessed by quality-of-life measures, early institutionalization, or health economics. UI is well known as a clinical triad for the diagnosis of normal pressure hydrocephalus (NPH). However, other than UI, NPH patients commonly have urinary urgency/frequency (overactive bladder: OAB), and less commonly, voiding difficulty. Fourteen percent of the patients (either women or men) have post-void residual > 100 ml. The most common urodynamic abnormality is detrusor overactivity (DO), which was noted in 95% of patients. The underlying pathophysiology for OAB/DO in patients with NPH seems to be decreased cerebral blood flow in the right frontal cortex, and to a lesser extent, altered basal ganglia function. Functional UI can overlap the OAB/DO due secondary to impaired cognition/initiative, immobility, or disturbed consciousness in this disorder. PMID- 18402071 TI - [Neuroimaging of idiopathic normal pressure hydrocephalus--current concepts]. AB - Neuroimaging plays an important role in the diagnosis and management of the idiopathic normal pressure hydrocephalus (iNPH). In addition to a ventricular dilatation with an Evans index of 0.3 or more, a tightness of the subarachnoid space in the high convexity/midline areas with a concurrent dilatation of the sylvian fissure and basal cistern is characteristic in iNPH. Voxel-based morphometry techniques can readily determine the validity of these findings. Periventricular hyperintensity, marked flow void in the aqueduct, and ventricular reflux and stasis of contrast agents are believed to have limited advantages in differential diagnosis and patient management. Cerebral blood flow measurement may help in discriminating iNPH from other disorders; however, partial volume effects due to disproportionate narrowing and dilatation of the subarachnoid space can cause substantial errors during post-processing and interpretation. PMID- 18402072 TI - [Development of shunt technology especially for idiopathic normal pressure hydrocephalus]. AB - Outcome of the treatment on hydrocephalus especially in idiopathic normal pressure hydrocephalus (iNPH) shows the better outcome after shunting operation according to the evolution of the shunt system and the better management of shunt complications, and also, comprehension of the pathophysiology of NPH. The history of shunt system is benefit for the understanding of the now available shunt system. The first generation system is emerge as the typical standard differential pressure valves. The second generation valves was developed for the management of overdrainage problems and siphoning control. Japanese guidelines of iNPH recommended the use of the programmable valve for shunt system. Now, various shunt system have been developed for the treatment of hydrocephalus. After millennium, the new third generation shunt systems that are consisted with programmable valve and anti-siphon devices, are now line up for the management of iNPH. The new recognition of NPH and the familiarity with these various shunt devices are enable to the new stage of the clinical sciene of the diagnosis and the management of iNPH. PMID- 18402073 TI - [Neural substrates underlying cognitive expertise]. AB - Recent behavioral and neuro-anatomical studies of cognitive expertise have suggested that that superior performance in memory experts is neither due to extraordinary general intelligence nor anatomical brain difference. Furthermore, functional neuro-imaging studies have revealed that expert performance in mental abacus and memory experts is not attributable to increased brain activities of the process that exists in non-experts or to hyper-activity in the prefrontal cortex. On the contrary, cognitive experts utilize brain areas that are not used by non-experts. These brain regions are directly related to exceptional knowledge and unique strategies that are acquired by experts through extensive practice. These neuro-imaging findings support psychological theories that cognitive expertise is achieved by developing extensive knowledge and novel cognitive strategies not used by non-experts. These lines of evidence suggest that ordinary people could become experts through appropriate practice. PMID- 18402074 TI - [Temporal order judgment in the brain]. AB - It is generally accepted that the brain can resolve the order of two stimuli that are separated in time by 20-50 ms, irrespective of their sensory modalities. This may support a traditional view that there is a single common decision mechanism of temporal order that compares arrival times of sensory signals of any modalities. However, the traditional view has been challenged recently in two ways. Firstly, it has been shown that human judgment of the temporal order is liable to change depending on our prior experiences, and in a modality specific manner. A constant tone-light pair presented repeatedly is judged as occurring simultaneously (lag adaptation), whereas opposite perceptual changes occur in judging the order of tactile stimuli delivered one to each hand. The latter change conforms to a Bayesian integration theory. Secondly, subjective temporal order has been shown to be inverted in some special conditions. Crossing the arms caused inverting of the temporal order of two tactile stimuli that were separated in time by 100-200 ms. The temporal order of visual and tactile stimuli separated by 50 ms was inverted when they were delivered just prior to the onset of a saccade. These recent results suggest that the mechanism of temporal order judgment is not so simple as previously hypothesized but would involve multiple processes whose results are later integrated to reconstruct a subjective order of events in time. PMID- 18402075 TI - [Clinicoradiological studies and frequency of incidental meningiomas on brain check-up]. AB - The purpose of this study is to evaluate clinico-radiological findings and to estimate frequency of incidental meningiomas detected by brain check-up (BC). A total of 12,258 healthy adults (8,515 men and 3,743 women) had BC, using physical check-up, brain magnetic resonance imaging (MRI) and MR angiography. Mean age (SD) was 52.6 (11.4) years, 52.7 (11.2) in men and 52.5 (11.8) in women. Duration of this study was between April 2004 and March 2007. BC subjects were divided into the first BC and the repeated BC group. The number of the first BC subjects was 5,056 (3,284 men and 1,772 women). In the repeated group defined that they had the first BC before April 2004 in our center, there were 7,202 subjects (5,231 men and 1,971 women). Fifteen subjects (7 men and 8 women) were diagnosed as meningiomas. Fourteen cases were detected by the first BC and one case by the repeated BC. The percentage of incidental meningiomas in the first BC group was calculated as 0.28, 0.18 in men and 0.45 in women. The sex ratio of women/men was 2.5. The mean age of those cases was 55.2 (10.1), 54.6 (12.9) in men and 55.8 (7.7) in women. Incidental rate of meningioma in the repeated BC group was calculated as 0.01% and 0.02% in men. No subjects had meningioma at age < or = 30 years. Asymptomatic meningiomas were seen in 14 cases (93.3 %) and they expected BC to ascertain healthy and normal brain by themselves. The size of tumors was ranged 10 to 46 mm and mean (SD) was 21.3 (10.2) mm. The locations of meningiomas were variable. Three cases had surgical removal. Twelve cases were under investigation on MRI. In middle-aged working subjects, incidental rates of asymptomatic meningiomas detected by BC are not uncommon in Japan. We should pay more attention to incidental meningioma when brain MRI is performed in 40-60 years women. PMID- 18402076 TI - [Memory impairment due to circumscribed infarct at the left dorsolateral prefrontal lobe]. AB - A 65-year-old man experienced an embolism at the left dorsolateral prefrontal region of the brain (the inferior frontal gyrus and a part of the middle frontal gyrus). He was a known case of diabetes mellitus and atrial fibrillation. On evaluation his Mini-Mental State Examination (MMSE) score was 26, indicating the absence of dementia. His chief complaint was episodic memory impairment. On the Wechsler Memory Scale-Revised (WMS-R), he had lower scores for general memory, visual memory and verbal memory. In particular, his verbal memory was markedly impaired compared to visual memory. Tests performed to assess working memory, such as evaluation on the WMS-R for attention/concentration in WMS-R and both parts A and B of the trail making test, revealed normal findings; this indicated that the patient had normal working memory. However, he complained of forgettingess of either phone numbers or routes, and this suggested the impairment of working memory in his daily life. In summary, he suffered from either episodic verbal memory or working memory impairment. His executive function test revealed a slight disturbance of set-exchange on the Wisconsin Card Sorting Test-Keio version. Such a case of memory impairment caused by a lesion in the left dorsolateral prefrontal lobe has rarely been reported. PMID- 18402077 TI - [Bickerstaff's brainstem encephalitis with positive findings on head MRI--case report]. AB - We report a case of Bickerstaff's brainstem encephalitis (BBE) with positive findings on head MRI. Abnormal lesions on head MRI were detected in approximately 30% cases of BBE. An 81-year-old female presented with disturbance of consciousness (Japan Coma Scale 200), ophthalmoplegia (downward deviation) and tetraplegia. Cerebrospinal fluid examination revealed mild pleocytosis 244 cells in 3 microl of CSF with normal glycorrhachia. Head MRI showed high-intensity abnormalities extending over the brain stem. The patient was diagnosed with BBE. It was thought that her symptoms were believed to arise from the lesion in reticular formation, rostral interstitial nucleus of the medial longitudinal fasciculus, and the pyramidal tract. At discharged after 1 month, she had no sequela. PMID- 18402078 TI - [Vertebral arteriovenous fistula as a complication of atlantoaxial transarticular screw fixation--case report]. AB - We report a rare case of a vertebral arteriovenous fistula that developed as a complication of atlantoaxial transarticular screw fixation. The patient was a 44 year-old male with a history of juvenile rheumatoid arthritis. He had undergone an atlantoaxial transarticular screw fixation for an atlantoaxial dislocation. At 2 months after the surgery, he complained of right-side tinnitus. A selective left vertebral angiography showed a high-flow arteriovenous fistula of the right V2 segment and occulusion of the right vertebral artery at the level of the C3 vertebral body. Endovascular embolization of the arteriovenous fistula was successfully performed using detachable coils. No deficits were observed after the treatment, and the tinnitus disappeared completely. Endovascular coil embolization is currently an effective and safe treatment for the vertebral arteriovenous fistula. PMID- 18402079 TI - [3D CT findings of biparietal thinning]. PMID- 18402080 TI - [One hundred books which built up neurology (15)--Charles Bell: Essays on the anatomy of expression in painting (1806)]. PMID- 18402081 TI - Rare case of myloglossus in Japanese cadaver: anatomical and developmental considerations. AB - The styloglossus is one of the three extrinsic muscles of the tongue; it enters the tongue from behind. During human dissection practice for dental students at the Showa University School of Dentistry in 2006, the authors observed anomalous muscle arising from the inner surface of the mandibular angle and inserting into the tongue bilaterally. This muscle was termed myloglossus by Wood. On the right side, the myloglossus was accompanied by a weak rudimentary styloglossus, but the styloglossus was absent on the left side. The hypoglossal nerve innervated the myloglossus bilaterally, and a small branch of mandibular nerve also entered the muscle on the left side. The styloglossus develops from the styloid process and the mandibular angle as a bicipital muscle, but the segment attached to the mandibular angle generally atrophies during development. The present anomalous muscle was thought to represent persistence of this muscle bundle arising from the inner surface of the mandibular angle. The innervation of the innermost part of the left myloglossus by the mandibular nerve was explained by persistence of a vestigial nerve supplying a small muscle bundle originating from the tensor veli palatini, which had fused to the myloglossus. PMID- 18402082 TI - Gross heart anatomy of Arctocephalus australis (Zimmerman, 1783). AB - Little research has been carried out on the gross visceral anatomy of the Otariidae, and the anatomical information for the southern fur seals, Arctocephalus spp., is scant. The aim of the present study was to describe the external and internal conformation, and the sanguineous irrigation of the heart of Arctocephalus australis. Twelve hearts of Arctocephalus australis were studied by simple dissection. In the right ventricle the trabeculae carneae were well developed and there were three or more papillary muscles. In the left ventricle there were two papillary muscles, subatrialis and subauricularis, attached to the parietal wall. There was also a great development of trabeculae carneae which occupied almost all of the ventricle, from the left atrioventricular valve up to the proximities of the expulsion route. A large quantity of muscular strands were found extending themselves between the trabeculae carneae, becoming more dense and forming a network when near the apex. The distribution of the branches of the coronary arteries was highly variable and no heart was similar to another one in this sense. In the majority of the hearts the subsinosal interventricular branch proceeded from the right coronary artery. It is concluded that there were many differences between the heart of the Arctocephalus australis and the heart of the domestic dog, contrary to what has been suggested for other genera of Otariidae. PMID- 18402083 TI - Study of the eye and lacrimal glands in experimental iron overload in rats in vivo. AB - A variety of syndromes leading to hemosiderosis in men cause ocular lesions. The purpose of the present study was to determine the distribution of iron and of transferrin in the eyes and lacrimal glands of rats in experimental hemosiderosis, so as to achieve a better understanding of the formation of the ocular lesions observed in patients with advanced hemosiderosis. In order to achieve hemosiderosis the rats were fed 3% (w/w) carbonyl iron or received i.p. or i.v. polymaltose iron. Hemosiderin deposits were detected in macrophages lying in the interlobular connective tissue of lacrimal glands, in the interstitial connective tissue of the choroid, in the ciliary body, in the iris and extracellularly in the sclera in all animals that received iron i.v. Also, scanty hemosiderin laden macrophages were found to a lesser degree in interstitial connective tissue of the choroid and in the interlobular connective tissue of lacrimal glands in animals that received iron i.p. No iron deposits were detected in the eye and lacrimal glands of control rats and in rats that were on an iron enriched diet. No transferrin was detected in the eye and in the lacrimal glands, neither in the control rats nor in the rats that received iron. Experimental iron overload leads to increased iron deposition in tissues of the eye and lacrimal glands, whereas no transferrin could be detected in the aforementioned organs. PMID- 18402084 TI - Morphological study of external oblique motor nerves and nuclei in cats. AB - In order to clarify the morphological features of peripheral motor nerves and motoneurons that innervate trunk muscles, the size distribution of external oblique (EO) peripheral motor fibers and motoneurons of the thoracic and the lumbar segments were examined. Histograms of the size distribution of EO motor fibers in peripheral nerves after ganglionectomy clearly had a bimodal distribution of small fiber groups and large fiber groups. It is very likely that small fiber groups correspond to gamma motor fibers and large fiber groups to alpha motor fibers. Gamma and alpha motor fiber groups were separated at 8-14 microm. The average diameter of the gamma and alpha motor fibers were different in each segment. The ratio of gamma and alpha motor fibers was approximately 1:2.0 in the thoracic segments and from 1:1.8 to 1:0.9 in the lumbar segments. Horseradish peroxidase was applied to the central stump of EO nerves, and the size distribution of EO motoneuron cell bodies in the thoracic and the lumbar spinal cords was examined. The size distribution of motoneuron cell bodies was bimodal in one cat (small and large motoneurons) and unimodal in three cats. When the ratio of small motor fibers to large motor fibers in peripheral nerves was applied to that of small motoneurons to large motoneurons, the separation of small and large motoneurons was approximately 40 microm. These results suggest that the morphological characteristics in peripheral nerves of trunk muscles are not reflected in motoneurons. PMID- 18402085 TI - Intersection patterns of human coronary veins and arteries. AB - Intersections between the coronary veins (CV) and arteries (CA) of 103 adult human hearts were mapped on the heart surface. Then the correlations of these intersection patterns to their localization were studied. Eight spots were selected where one of four major CV (anterior cardiac vein, middle cardiac vein, left posterior ventricular vein, and great cardiac vein) intersected with one of CA and their branches (right coronary artery, posterior interventricular branch, left posterior ventricular branch, circumflex branch, diagonal branch, and anterior interventricular branch). The great cardiac vein (GCV) ran beneath the anterior interventricular branch in 56 specimens out of 103, beneath the diagonal branch in 75 specimens out of 103, and beneath the circumflex branch in 36 specimens out of 103, while the other CV mostly ran over CA. The present observations suggest that the CV on the right side may be formed prior to CA, while the CV on the left side may be formed simultaneously with CA. PMID- 18402086 TI - Morphological study of the ansa cervicalis and the phrenic nerve. AB - Morphological features of ansa cervicalis and phrenic nerve were studied in 106 cadavers. Ansa cervicalis was located medial to the internal jugular vein in 63% (medial type) and lateral to the vein in 33.7% (lateral type). Ansa cervicalis was derived from a combination of C1-C4 spinal segments, with C1-C3 being the most frequent pattern (87.5%). In >60% the ansa was bilaterally symmetrical. The distribution of medial and lateral types was equal on left and right sides of the body. The segmental composition of the inferior root was higher in the medial type and also on the left side of the body. In the lateral type the branches that formed the inferior root frequently (75%) formed a common trunk before joining the superior root, but in 74.8% of the medial type they joined the superior root independently. The phrenic nerve was derived from C4 and C5 in 52%. The C4 segment was present in the phrenic nerve in all cases except one. Additional phrenic components that pass anterior to the subclavian vein were defined as accessory phrenic nerves and found in 28.7%, while those passing posterior to the same vein were defined as secondary phrenic nerves (19.8%). Most of the accessory phrenic nerves contained a C5 segment and the nerve to subclavius was the commonest source. Various relationships between the ansa cervicalis and the phrenic nerve are investigated and, based on these findings, two separate classifications for the two nerves are suggested. PMID- 18402087 TI - Clinical relevance of palmaris longus agenesis: common anatomical aberration. AB - Palmaris longus muscle, although of little functional use to the human upper limb, assumes great importance when used as a donor tendon for transfer or transplant. The variability in the prevalence of palmaris longus agenesis among various ethnic groups has been established, and the surgeon's awareness of the prevalence in a population or ethnic group is desirable. The prevalence of palmaris longus agenesis has, to the best of the authors' knowledge, not been reported in Indian patients. Five hundred Indian patients were examined for the presence or absence of palmaris longus tendon, using the conventional test for presence of palmaris longus. The prevalence and pattern of palmaris longus agenesis was analyzed statistically and any difference in prevalence or pattern of palmaris longus agenesis with regard to body side or sex was looked for. All statistical analysis was done using SPSS (version 12). chi2 test was used to analyze the association of agenesis with limb laterality and sex. The prevalence of palmaris longus agenesis was found to be 17.2% (8% bilateral and 9.2% unilateral). The prevalence of agenesis was significantly more common on the left side. Male subjects had a greater likelihood of unilateral agenesis, while female subjects were more likely to have bilateral agenesis. That prevalence of palmaris longus agenesis is race dependent is reaffirmed in the present study. Although the prevalence of palmaris longus agenesis in Indian patients was found to be much higher than the reported average prevalence in an Asian population, this tendon can still be counted on by surgeons treating Indian patients for use as a donor tendon, which will be present in a vast majority of Indian patients. PMID- 18402088 TI - Right ligamentum teres joining to the right branch of the portal vein. AB - Ligamentum teres joining to the right branch of the portal vein in a 79-year-old Japanese male cadaver was noted during student dissection at Kumamoto University in 2004. The ligamentum teres entered the liver along the left side of the gallbladder fossa. The quadrate lobe was not distinguished from the left lobe in the visceral surface. When the liver parenchyma was removed by tearing off to expose the branches of the portal and hepatic veins, it was clarified that the ligamentum teres unusually joined to the bifurcation of the upper anterior and lower anterior branches of the right branch of the portal vein. The ligamentum teres is the remnant of the umbilical vein working throughout fetal life. Initially a pair of the umbilical veins entered the sinus venosus. During the fourth and fifth weeks they connect to the hepatic sinusoids, which become the portal and hepatic veins, and the parts entering the sinus venosus of both umbilical veins disappear. By the eighth week, as all remainder of the right umbilical vein disappears, the left umbilical vein is the only one to carry blood from the placenta to the liver. It results in the ligamentum teres joining to the left branch of the portal vein. However, in the present case it is thought that the right umbilical vein remained instead of the left one for some reason, and it then became the right ligamentum teres joining to the right branch of the portal vein. PMID- 18402089 TI - Nerve fiber analysis on the so-called accessory subscapularis muscle and its morphological significance. AB - A rare muscular anomaly, so-called accessory subscapularis muscle, was found in the left axillary fossa of a 95-year-old male cadaver during a student dissection practise. The muscle arose near the lateral margin of the scapula from the surface of the subscapularis muscle and ran upward to fuse with the capsule of the shoulder joint via a tendon. It measured 1.0 cm in width, 7.0 cm in length and 1.5 mm in thickness, and was separated from the underlying subscapularis muscle by the axillary and inferior subscapular nerves. Macroscopically, the anomalous muscle received its nerve supply from a branch arising from the lower root of the radial nerve near the origin of the thoracodorsal nerve and entered the muscle from its ventral surface. Nerve fiber analysis showed that the supplying nerve originated from fibers of the dorsal element of C7 immediately cranial to the thoracodorsal nerve. These findings indicate that the present anomalous muscle might be close to the formation of the latissimus dorsi muscle in its derivation rather than the subscapularis muscle. PMID- 18402090 TI - Professionalism. PMID- 18402093 TI - In vitro evaluation of the influence of air abrasion on detection of occlusal caries lesions in primary teeth. AB - PURPOSE: The purpose of this study was to assess the influence of cleaning pits and fissures with an aluminum oxide air abrasion system on the detection of occlusal caries in primary teeth using loser fluorescence (LF) and visual examination. METHODS: The sample comprised 65 pit and fissure sites on extracted primary teeth suspected to be carious. The sites were submitted to 2 visual examinations (examiner JAR) and 2 LF readings (examiner TMV). Next, the occlusal surfaces were air-abraded and re-examined thereafter using both methods. The teeth were sectioned, and the histological analysis of the sites with a stereoscopic magnifying lens at X32 magnification was used as the gold standard. RESULTS: Cohen's kappa statistic for LF and visual examination were, respectively, 0.282/0.884 before and 0.896/0.905 after air abrasion. LF showed a sensitivity of 0.28 increasing to 0.49 and a specificity of 0.50 increasing to 0.92. Visual examination showed sensitivity of 0.78 and specificity of 0.73. Both increased after air abrasion. CONCLUSION: The findings suggest that cleaning pits and fissures with aluminum oxide air abrasion increased the accuracy of LF and visual examination for detection of occlusal caries in primary teeth. PMID- 18402092 TI - The relationship between acute otitis media and the anatomic form of the hard palate. AB - PURPOSE: Acute otitis media (AOM) is a serious health concern for millions of children. This study was conducted to determine the relationship between AOM and the shape of the hard palate. METHODS: Intraoral examinations were performed on 175 pediatric dental patients ages 4-6. The palatal height was classified as high, medium or low. The accompanying parent completed a survey regarding their child's age, gender, overall health, history of AOM and presence of known risk factors for AOM. RESULTS: Eighty five percent of the total sample had a positive history for AOM, with 76% experiencing AOM prior to age 1. High palatal vault was a significant finding in children that experienced AOM before age 1 (odds ratio 3.49). Logistic regression analysis revealed that children with high palatal vaults underwent tube placement more often than the rest of the study population (odds ratio 2.49). CONCLUSIONS: Our study revealed a relationship between the presence of high palatal vaults and early, recurrent acute otitis media in young children. Recognition of high palatal vault in children suffering from AOM, in the presence of other known risk factors, may guide health care professionals to identify an otitis media prone patient and aid in prevention of recurrence. PMID- 18402094 TI - Occlusal fossae depth of permanent first molars assessed by visual examination: an in vitro approach. AB - PURPOSE: Sealant placement commonly is employed in molars with deep fissures to protect the teeth from occlusal caries. Some have advocated the some preventive approach for deep occlusal fossae in permanent molars. The decision of whether or not to place a sealant depends largely on the dentist's assessment of the depth of the occlusal fossae. The purposes of this in vitro study were to: (1) evaluate the morphology and determine the actual depth of occlusal fossae from newly erupted permanent first molars; (2) compare them with depth assessments obtained from visual examination, and (3) characterize how pediatric dentists look at the occlusal surfaces to assess fossae depth. METHODS: Fossa depth of 48 intact permanent molars was measured by a computerized micromeasurement system (Michigan Laser Digitizer) and compared with the depth assessed during a bench-top examination conducted by a panel of pediatric dentists. RESULTS: The computerized and visual measurements were positively and significantly correlated (r=0.64, P<.001). Agreement as measured by the Kappa statistic was moderate (k=0.46; 95% confidence interval=0.22-0.71; P<.001). CONCLUSION: Pediatric dentists' perception of fossa depth in permanent molars correlates moderately well with the actual fossa depth. PMID- 18402095 TI - Effectiveness of primer and bond in sealant retention and caries prevention. AB - PURPOSE: The purpose of this clinical trial was to test the use of a primer and bonding agent to increase the retention of a fissuresealant in a group of Kuwaiti children. METHODS: In 78 6- to 9-year-old children who required sealants in all 4 permanent frst molars, sealants were placed in 2 teeth using a primer and bond (3M Scotch Bond) and in 2 teeth without primer and bond. Sealant retention was evaluated 1 and 2 years later. Sealant retention was scored as: (1) complete retention; (2) partial loss of sealant; and (3) complete loss of sealant Caries was scored for those teeth in which the sealant was partially or completely lost RESULTS: At the 2-year examination, in teeth sealed with primer and bond (a) 64% of the sealants were completely retained; (b) 23% were partially lost; and (c) 13% were completely lost. In teeth sealed without primer and bond: (a) 68% of the sealants were completely retained; (b) 20% were partially lost; and (c) 12% were completely lost There was no difference in sealant retention (P=.22) and caries (P=.56) in teeth sealed with and without bond. In multivariate analyses after controlling for age, gender, tooth surface, and arch, no differences in sealant retention and caries were seen. CONCLUSION: If a proper technique is used in sealant placement, primer and bond does not enhance sealant retention. PMID- 18402096 TI - Marginal microleakage of a sealant applied to permanent enamel: evaluation of 3 application protocols. AB - PURPOSE: The purpose of this in vitro study was to investigate and compare the microleakage-occurring after the placement of a light-curing sealant-to unground permanent enamel which had been previously conditioned using 3 different application protocols. METHODS: Sixty-three molars were randomly distributed in 3 different groups according to the application protocol of the sealant: (1) conditioning of the enamel with 38% phosphoric acid (group 1); (2) conditioning with phosphoric acid and single-bottle dentin bonding agent (group 2); or (3) conditioning with a self-etching adhesive (group 3). Samples were: (1) thermocycled; (2) stained; (3) sectioned; and (4) examined for marginal microleakage. RESULTS: No dye penetration was noted for 25%, 59%, and 40% of cases for groups 1, 2, and 3, respectively. CONCLUSIONS: Placement of a bonding agent layer prior to the sealant allows significantly less microleakage than the traditional conditioning of enamel with phosphoric acid alone. The self-etching adhesive used here seems to be an attractive alternative to the acid-etch and adhesive technique for sealant application in young children, since it would simplify the procedure. Clinical trials should be performed to assess the performance of these products before definitive conclusions can be formulated. PMID- 18402097 TI - Light-cured calcium hydroxide vs formocresol in human primary molar pulpotomies: a randomized controlled trial. AB - PURPOSE: The purpose of this prospective study was to compare light-cured calcium hydroxide (Ca(OH)2) with diluted formocresol (FC) for its success as a primary molar pulpotomy medicament METHODS: Selection criteria included at least 2 matching, asymptomatic, contralateral primary molars requiring vital pulpotomies. Matched teeth in each patient were randomized to receive either Ca(OH)2 or FC as a pulpotomy medicament. All teeth were restored with prefabricated metal crowns. Twenty patients (34 pairs of teeth) were followed clinically and radiographically for > or =1 year. Two blinded, standardized, and calibrated examiners evaluated and scored each radiograph for signs of pathology, based upon a modified scale previously proposed. RESULTS: Findings were grouped in: (a) 0 - 6; (b) 7 - 12; and (c) 13 - 24 month intervals. Radiographic scoring favored the FC group of the 7- to 12- and 13- to 24-month intervals (P<.05). Clinical success was similar for Ca(OH)2 (94%) and FC (97%) at 12 months or less. Beyond 12 months, clinical success varied more (Ca(OH)2=84%, FC=97%), but not significantly (P=.08). Combined success rates were lower for Ca(OH)2 (56%) than FC (94%). CONCLUSIONS: Light-cured calcium hydroxide does not appear to be a viable alternative to diluted formocresol as a pulpotomy agent. PMID- 18402098 TI - A survey of primary tooth pulp therapy as taught in US dental schools and practiced by diplomates of the American Board Of Pediatric Dentistry. AB - PURPOSE: The purpose of this study was to repeat a 1997 survey of current pulp therapy practice. METHODS: The directors of dental school predoctoral pediatric dentistry programs (N=56) and board certified pediatric dentists (N=1200) were surveyed in 2005. RESULTS: More dental schools (83%) taught indirect pulp therapy (IPT) compared to 1997. Significantly more used glass ionomer for IPT with most dental schools and diplomates not re-entering a tooth after IPT. Over 30% of schools and diplomates do direct pulp cops using glass ionomer. For pulpotomy, diluted formocresol usage decreased in dental schools (54%) while ferric sulfate significantly increased (24%) and full strength remained at 22%. Shorter placement of pulpotomy medication was noted and ZOE alone the preferred base. Pulpectomy was advocated by 85% of 2005 schools and diplomates with ZOE filler use decreasing while iodoform/calcium hydroxide filler use increasing. CONCLUSIONS: More pediatric dentists are using glass ionomer for IPT and direct pulp capping, and there was a trend away from the use of 1:5 diluted formocresol with more using ferric sulfate for pulpotomy. For pulpectomy, most use ZOE but iodoform pastes and calcium hydroxide have increased in usage since 1997 Disagreements continue concerning when to use certain pulp therapies and some directors and diplomates did not follow the AAPD guidelines. PMID- 18402099 TI - Caries risk assessment practices among Texas pediatric dentists. AB - PURPOSE: The purpose of this study was to determine current caries risk assessment (CRA) practices of Texas pediatric dentists. METHODS: A 20-question survey was sent to all 204 active members of the Texas Academy of Pediatric Dentistry. The mailing list was obtained from the Texas Academy of Pediatric Dentistry roster. RESULTS: The response rate was 62% (127/204). Eighty-three percent of respondents reported that parents are receptive to education about caries risk factors. Socioeconomic status was unrelated to receptivity. Ninety three percent of respondents reported that they are actively involved in CRA. Thirty-eight percent of those responding reported performing CRAs on greater than 76% of their patients; 9% reported no CRA use. Forty percent of respondents reported that they assess caries risk but do not document the caries risk status. Seventy percent of respondents reported a desire for more CRA education. No significant differences were found between: (1) year o fgraduation; (2) practice type; or (3) payer sources received and CRA practices. CONCLUSIONS: A need exists for more comprehensive caries risk assessment practices and increased documentation of caries risk status by Texas pediatric dentists. The continuing education of dental professionals in caries risk assessment is essential. PMID- 18402100 TI - Dental needs and status of autistic children: results from the National Survey of Children's Health. AB - PURPOSE: The purpose of this study was to assess the oral health status and dental needs of a nationally representative sample of 1- to 17-year-old children with or without autism. METHODS: In the 2003 National Survey of Children's Health, parents reported their child's oral health status and needs. The condition of the child's teeth, demographics, time since last dental visit, and dental needs were assessed in autistic children (N=495) and nonautistic children (N=95,059). For a subset of children with reported fair or poor teeth, specific problems about their dentition were assessed for autistic children (N=69) and nonautistic children (N=7,002). Weighted percentages and chi-square statistics were calculated. RESULTS: According to parents, 69% of nonautistic children and 52% of autistic children had their teeth in excellent or very good condition (P <.001). The dental status of children with autism and without autism, identified with fair or poor teeth, was comparable. CONCLUSIONS: Overall, parents of US autistic children were more likely to report their children's dentition to be in fair or poor condition than parents of US nonautistic children. Children with or without autism who had fair or poor teeth are faced with similar dental problems. PMID- 18402101 TI - Stainless steel crown aspiration during sedation in pediatric dentistry. AB - Foreign body aspiration (FBA) causes death in more than 300 children every year in the United States. Morbidity and mortality are increased in children due to narrow airways and immature protective mechanisms. Factors to consider in pediatric dentistry are: (1) the patient's age and behavior; (2) presence and extent of disability; (3) local anesthesia; (4) body positioning; and (5) loose teeth. FBA requires prompt recognition and early treatment to minimize potentially serious and sometimes fatal consequences. The purpose of this case report was to describe the aspiration of a stainless steel crown in a 5-year-old boy during conscious sedation. It also discusses how a prompt and accurate diagnosis, early referral, and immediate treatment helped prevent serious complications. PMID- 18402102 TI - Management of an ectopically erupting permanent mandibular molar: a case report. AB - Ectopically erupting permanent first molars occur in approximately 3% to 4% of children, and presentation in the mandible is very rare. When the malocclusion warrants a nonextraction treatment plan in the mandible, early correction of an impacted ectopic molar with space regaining is appropriate. The purpose of this case report was to describe and document the correction of a severely impacted permanent mandibular left first molar in the early mixed dentition along with long-term results. A bilateral lingual arch was placed on the primary mandibular second molars with a Halterman-type extension in conjunction with a bonded button on the occlusal surface of the impacted permanent molar and chain elastic Early space regaining allowed future nonextraction treatment. PMID- 18402103 TI - Sequential supernumerary teeth in nonsyndromic patients: report of 3 cases. AB - Supernumerary tooth/teeth (ST) are a well recognized clinical phenomenon defined as teeth that are additional to the normal complement. The purpose of this report was to describe 3 patients who developed multiple sequential ST. All patients were boys between 8 and 12 years old without significant medical history. In 2 cases, the ST were discovered while investigating the nonexfoliation of primary teeth, whereas in the other they were an occasional finding on a radiograph taken for orthodontic purposes. All patients were kept under close follow-up ofter treatment. The 3 patients collectively developed a total of 17 ST with 9 found in a single patient, over a 5-year period. This article demonstrates the importance of clinical and radiographic follow-up of ST patients, regardless of age, family history, or systemic condition. PMID- 18402104 TI - The effects of general anesthesia legislation on operating room visits by preschool children undergoing dental treatment. AB - PURPOSE: The purpose of this study was to examine the impact of state-level general anesthesia (GA) legislation on operating room visits for the treatment of dental caries on preschool-aged children. METHODS: The North Carolina Ambulatory Surgery Discharge Database was used to observe GA visits for fiscal years (FY) 1997 to 2001. A pretest/post-test design with concurrent comparison groups was used for 2 analyses: (1) all children treated for dental caries were compared to those treated for otitis media; and (2) those whose treatment for dental caries was reimbursed by Medicaid were compared to those whose treatment for dental caries was not reimbursed by Medicaid. RESULTS: In the prelegislation period (FY 1997 and 1998), there were 3,857 GA visits for dental core and 21,038 for otitis media. Postlegislation (FY 2000 and 2001) dental visits increased to 5,511(43%), and otitis media visits increased to 22,279 (6%)-a statistically significant difference (P<.05). Before the legislation, there were 1,370 non-Medicaid dental visits and 2,487 Medicaid dental visits. Non-Medicaid and Medicaid dental visits postlegislation increased to 2,195 (60%) and 3,316 (33%), respectively. This difference was significant (P<.05). CONCLUSIONS: General anesthesia legislation resulted in an increase in access to care for children needing dental care in North Carolina. PMID- 18402105 TI - [Characteristics and biomechanical mechanism of riding injuries in accidents of bicycles collided by motor vehicles]. AB - OBJECTIVE: To study the characteristics and biomechanical mechanism of riding injuries involving bicycles collided by motor vehicles. METHODS: The real traffic accident cases of bicycles collided by motor vehicles, including the information of scenes, bicycles, motor vehicles, rider wounds and traffic directions, were collected. Retrospective method was used to study these riding injuries. In addition, typical cases were selected to simulate traffic accident courses with computer simulation software, and the dynamic data like acceleration, force, moment were cxtracted to compare with those in the real cases. RESULTS: There were no difference of occurring frequency between cases with or without riding injuries, as well as between one-side-collision and front- or back-collision. The riding injuries seemed less in accidents involving large-scale vehicles. The frequency of riding injuries increased with vehicle speed. The wound location was low on collision side and high on opposite. CONCLUSION: Analysis of riding injury characteristic in traffic accidents and their biomechanical mechanism would be helpful for estimation of traffic manner. PMID- 18402106 TI - [Estimation of postmortem interval with multivoxel-voxel proton 1H-MR spectroscopy]. AB - OBJECTIVE: The purpose of our study was to evaluate the postmortem interval with multi-voxel 1H-MR spectroscopy. METHODS: Twelve healthy rabbits were studied and the quantities of N-acetylaspartate, total choline, phosphocreatine and creatine were measured by 1H-MR spectroscopy after death at 0.5, 1, 2, 4, 6, 8, 12, 16, and 24 h. RESULTS: The levels of Naa/Cr and Naa/Ch decreased following death, while the level of Ch/Cr increased initially and then decreased following death. CONCLUSION: Multi-voxel proton MR spectroscopy for Naa/Cr and Ch/Cr metabolic ratio could be used in future postmortem interval studies. PMID- 18402107 TI - [Effects of serum from crush injury rats on vascular endothelial cell apoptosis and their potential mechanism]. AB - OBJECTIVE: To investigate the effects of serum from crush injury rats on vascular endothelial cell apoptosis and their potential mechanism. METHODS: Bovine aorta endothelial cells were cultured in vitro and the effects of serum from crush injury rats on cell apoptosis and intracellular calcium concentration ([Ca2+]i) were observed. Meanwhile, the levels of rat blood plasma endothelin-1 (ET-1) and atrial natriuretic peptide(ANP) were measured. RESULTS: Compared with normal rat serum treatment, the cell apoptosis rate decreased from (8.26+/-1.75)% to (2.75+/ 0.90)%, while the concentration of [Ca2+]i increased from (96.98+/-3.95) to (118.79+/-3.22) nmol/L in serum from crush injury rats, respectively. The concentration of ET-1 and ANP increased significantly in crush injury rat serum. CONCLUSION: Serum from crush injury rats could inhibit apoptosis of the vascular endothelial cells. These effects may be related to increased level of [Ca2+]i mediated by ET-1 and ANP. PMID- 18402108 TI - [Application of immunofluorescence and sandwich ELISA with double-antibodies in detection of human rabies]. AB - OBJECTIVE: To explore the application of immunofluorescence and sandwich ELISA with double-antibodies in detection of human rabies. METHODS: The cerebrum, cerebellum, brainstem, and hippocampus of four patients died of rabies identified by clinical diagnosis were collected and kept in freezer at -70 degrees C or in formaldehyde solution separately. The rat brain tissue infected by CVS strain of rabies virus was used as a positive control and the brain tissue of a patient died of acute pancreatitis was used as a negative control. RESULTS: Rabies virus was detected in the tissues kept in freezer at -70 degrees C and the positive control but was not detected in the tissues kept in formaldehyde solution and the negative control. CONCLUSION: Immunofluorescence and Sandwich ELISA with double antibodies could be used in detection of human rabies. The samples should be kept in deep frozen temperature condition instead of in formaldehyde solution. PMID- 18402109 TI - [Study on development of hand-wrist bone in 14 year-old adolescents I: bone age standard for boys]. AB - OBJECTIVE: To study the bone development in adolescents in China, and establish forensic skeletal age standard for estimation of the criminal responsible age in 14 years old male adolescents. METHODS: One hundred and three healthy boys aged from 13 years and 9 months to 14 years and 3 months in Lengshuijiang, Hunan, China were enrolled in this study. X-ray films of their left hand-wrist were taken dorsaventrally, and 15 selected sites on the wrist X-ray films were examined and measured under standard condition. The development grade was confirmed by maximum percentage to establish a method to estimate the bone age, which was then checked by single blind method. RESULTS: Development grade of the first metacarpal bone and the first phalanx IV was partially closed (less than 1/3), but the rest was non-closed. The cross diameters of all the osteoepiphysis were wider than that of all the diaphysis. The development grade of the fourth metacarpal bone and the fifth metacarpal bone as well as the adjacent third phalanx were different in both younger age group (<14 years, mostly non-closed) and the older age group (>14 years, mostly closed). The confirmed accuracy (in 30 boys) was 80%. CONCLUSION: Our study seems to be feasible in establishing the estimation standard for skeletal age determination using developmental metaphysis and anthropometry. The standard may be particularly helpful in forensic practice for estimation of accurate criminal responsible age in adolescents. PMID- 18402110 TI - [Stature estimation from upper extremity long bones by digital radiography]. AB - OBJECTIVE: To obtain regression formula for estimation of stature in Han population in Sichuan Province from length of the upper extremity long bones by digital radiography. METHODS: The statures of 365 healthy adults and digital radiographs of their right upper extremity long bones were measured. All statistical dispositions were done in SPSS including description and regression analysis. RESULTS: Twenty-seven simple regression and 3 multiple regression formulae with statistic significance in estimation of the stature from the sum of the length of the upper extremity long bones were established. Regression coefficient was higher in ulna than in radius, in male than in female, and was more accurate in multiple regression formulae than in simple regression formulae. CONCLUSION: Digital radiograph measurement of the length of the upper extremity long bones seems to be a simple and practical method to estimate human stature in forensic practice. PMID- 18402111 TI - [The effects of heroin on intracellular free Ca2+ of rat myocardium]. AB - OBJECTIVE: To observe the effects of heroin on intracellular free Ca2+ in rat myocardium. METHODS: The effects of heroin on intracellular free Ca2+ were observed in cultured neonatal rat myocardium by measuring intracellular free Ca2+ concentration using calcium fluorescent probe Flou-3/AM and laser scanning confocal microscope. RESULTS: Different doses and concentrations of heroin appeared to have different effects on intracellular free Ca2+ concentrations, with a dosage dependent short linear increase in the fluorescence intensity (i.e., Ca2+ concentration) leading to [Ca2+]i peak. CONCLUSION: Heroin could affect concentrations of [Ca2+]i in myocardium and its dosage related effect needs further investigation. PMID- 18402112 TI - [The role of dopamine D3 receptor in the amphetamine-induced conditioned place preference in mice]. AB - OBJECTIVE: To study the role of dopamine D3 receptor involved in the amphetamine induced conditioned place preference (CPP) in mice. METHODS: The CPP was observed in D3 receptor knock-out (D3RKO) mice and C57BL/6 wild-type control mice after administration of amphetamine. The data were analyzed with a two-way ANOVA using the SPSS 13.0 software. RESULTS: D3RKO mice showed a significant amphetamine induced CPP (P<0.001), compared with the ones administered with saline in C57BL/6 control mice. CONCLUSION: The results indicate that amphetamine can produce significant CPP in dopamine D3 receptor knock-out mice, suggesting that amphetamine-induced addiction can be inhibited by dopamine D3 receptor. PMID- 18402113 TI - [Application of spiral CT 3D reconstruction of rib fracture in clinical forensic practice]. AB - OBJECTIVE: To assess the clinical application of spiral CT 3D reconstruction in forensic diagnosis of rib fracture. METHODS: Thirty-five cases of rib fractures were collectively studied by X-ray, regular CT scan, and adoptive 3D reconstruction with spiral CT. RESULTS: Clear and stereoscopic images of rib fractures without interference of the surrounding soft tissue were obtained in all of 35 cases examined. CONCLUSION: Spiral CT 3D reconstruction could be a valuable means in forensic diagnosis of rib fracture. PMID- 18402114 TI - [The relationship between age-related stress distribution and fracture incidence of the mandible]. AB - OBJECTIVE: To analyze age-related mandible stress distribution due to midline force. METHODS: Mandibles of children, adults, and elderly individuals were scanned by spiral CT to establish three-dimension imaging models with mesh elements by MIMICS software and HYPERMESH software. The mandible stress distribution was analyzed using ANSIS software. RESULTS: There was no significant difference in mandible stress distribution in various age groups with the greatest stress distribution (Von Mises) present at the mandible angle. Although there was stress present at the mandible neck in adults, no such mandible neck stress was found in children and elderly individuals. CONCLUSION: Mandible stress distribution is closely related to the incidence of fracture in various age groups, i.e., more stress and more fracture. PMID- 18402115 TI - [Age-dependent mitochondrial DNA 4977bp depletion in human skeletal muscle]. AB - OBJECTIVE: To explore depletion of human mitochondrial DNA 4977-bp and its relation with aging. METHODS: Total DNA (nuclear and mtDNA) was extracted from 100mg muscle tissue. UV light illumination of ethidium bromide-stained PCR products was used to study the depletion of mtDNA (wild-type or mutant). RESULTS: The proportions of mtDNA depletion in human skeletal muscle could be determined. The frequency of mtDNA 4977-bp depletion in different age groups (0-9, 10-19, 20 29, 30-39, 40-49, 50-59, 60-69, 70-79, 80-89, 90-99) was: 0%, 0%, 0.003%, 0.011%, 0.015%, 0.033%, 0.038%, 0.062%, 0.069%, and 0.091%, respectively. CONCLUSION: Our findings suggest that the frequency of the mtDNA4977 depletion in human skeletal muscle increases with age. It might be useful for human age estimation. PMID- 18402116 TI - [Determination of mexiletine in human blood by liquid chromatography-tandem mass spectrometry]. AB - OBJECTIVE: To establish a liquid chromatography-tandem mass spectrometry (LC MS/MS) method for detection of mexiletine by liquid chromatography tandem mass spectrometry. METHODS: After simple protein precipitation of the blood sample with acetonitrile, the organic solvent layer diluted with LC mobile solvent was separated by Allure PFP Propyl column, confirmed and quantified by MS/MS in the multi-reaction monitoring (MRM) mode via positive electrospray ionization. RESULTS: Mexiletine and naloxone (internal standard) got ideal resolution under the selected analytical condition. The correlation coeficient of linear calibration curve was over 0.9999 within the mexiletine concentration range 0.02 10 microg/mL. The relative standard deviations were under 10% for intra-day and under 15% for inter-day, and the detection limit was 0.01 microg/mL. CONCLUSION: The established LC-MS/MS method is simple, rapid, sensitive, unaffected by matrix effect and appropriate for detection of mexiletine in blood in the field of therapeutic drug monitoring and forensic toxicology. PMID- 18402117 TI - [Distribution of tryptophan hydroxylase gene-T457C locus polymorphism in Han ethnic group in northern China]. AB - OBJECTIVE: To study the distribution of the tryptophan hydroxylase (TPH) gene T457C locus polymorphism in Han ethnic group in northern China and to find its applicable value in forensic science. METHODS: Genomic DNA samples, extracted from 180 unrelated individuals in northern Chinese Han population, were analyzed by PCR-RFLP. RESULTS: The discrimination power (DP) value and the power of exclusion (PE) value of the TPH gene-T457C locus were 0.624 and 0.187, respectively. The allele frequency showed significant difference from that of French people (P=0.04). CONCLUSION: Polymorphism of the TPH gene-T457C locus could show ethnic and regional differences. It has a potential to be used in forensic science. PMID- 18402118 TI - [Recent advances in venous air embolism]. AB - Air embolism, a life-threatening complication of medical procedure, is a frequently encountered challenge in the forensic practices. It can be easily missed due to ignorance by forensic examiner or it could be difficult to be identified due to prolonged storage of the cadaver. This article reviews the etiological factors, pathophysiological changes, clinical manifestation, diagnosis, and the medicolegal identification of air embolism. The authors suggest that modern imaging techniques including echocardiogram, computer tomography, and magnetic resonance imaging play animportant role in the clinical diagnosis and forensic identification of air embolism. PMID- 18402119 TI - [The application of cerebral angiography in the forensic postmortem examination]. AB - As a "Gold Standard" in clinical diagnosis of cerebrovascular disease, cerebral angiography also plays an important role in forensic postmortem examination. The key roles of identifying the site of cerebrovascular disease and the cause of death as well as providing guidance for autopsy and postmortem sampling and the broad future application of cerebral angiography in forensic practice are reviewed. PMID- 18402120 TI - [Recent progress in epilepsy in neuropathology]. AB - Epilepsy is a common cerebral disease, and may cause sudden death. Although electric activity study of epileptic brain had been emphasized in the past, the neuropathological study of epilepsy has become a main focus in clinical and forensic medicine recently. This article reviews the recent progress in neuropathology of epilepsy including developmental disorder, abnormal tumoral proliferation, hippocampal sclerosis, dual pathological alteration, and mossy fiber sprouting. Its significance in forensic medicine, particularly for the diagnosis of sudden unexpected death in epilepsy, is discussed. PMID- 18402121 TI - [The application of palynology in forensic medicine]. AB - Palynology, one science of plant's pollen and spores, has been proven to be a new frontier discipline. Because of the characteristics of pollen and spores, such as small size, light weight, large amount, and difficult to be found, they can leave physical evidence and provide new clues to solve a case. Therefore palynology has a good prospect for practical application in forensic medicine. The paper intends to analyze the advantage and limitation of palynology in forensic medicine by reviewing its general characteristics, classification, morphology, and disseminating circadian rhythm. We hope to provide some reference to apply palynology in forensic medicine. PMID- 18402122 TI - Successful interventional dilatation of a complicated stricture of the anastomotic colon: report of a case. AB - Colonic anastomotic strictures are usually secondary to ischemia, leakage, inflammation, or hemorrhage at the anastomotic line. Several techniques for stricture dilation including endoscopic and radiologic dilation have been reported to avoid surgery. However, if colonic strictures are more than 1 cm long and the diameter is <5 mm, surgery is usually necessary. We describe the combined use of bougie, stent, and endoscopic techniques in successful dilation of a complicated anastomotic stricture of the colon, with a length of 5 cm with angulation and a diameter of <5 mm. PMID- 18402123 TI - Oral fluoropyrimidines may not reduce the risk of postoperative recurrence in colorectal cancer associated with mesenteric lymph node metastasis. AB - To clarify the efficacy and problems of postoperative adjuvant chemotherapy using oral fluoropyrimidines, the clinicopathological data of 307 colorectal cancer patients treated with or without postoperative chemotherapy were analyzed retrospectively. Patients in the chemotherapy group (n=188) who underwent curative resection were followed by administration of oral fluoropyrimidine. The other 119 patients underwent surgery alone. The disease-free survival rates were compared between the two groups. The disease-free survival rate in the chemotherapy group was significantly higher than that in the surgery alone. However, no significant difference in disease-free survival rate was found for those with tumors that were associated with mesenteric lymph node involvement and tumors with a high grade of lymphatic invasion or high grade of venous invasion. Postoperative adjuvant chemotherapy using oral fluoropyrimidines such as UFT (litegafur +4:uracil) and 5'-DFUR (doxifluridine) might not reduce the risk of recurrence in colorectal cancer with mesenteric lymph nodes involvement. PMID- 18402124 TI - Emergency surgery for complicated colorectal carcinoma: a comparison of older and younger patients. AB - The outcomes after emergency surgery for complicated colorectal cancer of 54 patients >70 years of age (group 1) and 53 patients < or =70 years of age (group 2) treated between 1991 and 2002 were compared. Forty-two patients had obstructions in group 1 and 41 patients had obstructions in group 2. Twelve patients had perforations in each group. Significantly higher ASA grading and APACHE II scores were seen in group 1. The overall and major morbidity and mortality rates in groups 1 and 2 were 81.5% and 58.5% (P = 0.009), 42.6% and 24.6% (P = 0.04), and 27.8% and 1.9% (P = 0.000), respectively. It was concluded that the outcome in older patients who underwent emergency surgery for complicated colorectal carcinoma was poor compared with that of younger patients. Careful preoperative assessment of the patient's general condition and intensive treatment after surgery in those with a poor physical status are recommended. PMID- 18402125 TI - Papillary carcinoma arising in a thyroglossal duct cyst; two case reports and review of the literature. AB - We present in this report two cases of papillary carcinoma arising in a thyroglossal duct cyst. The first case was a 32-year-old female patient who presented with a neck mass of 5 years' duration that had recently increased in size. The patient was otherwise asymptomatic. The second patient was a 41-year old male patient who presented with a submental mass that had been growing over the previous several months. Associated symptoms included local symptoms such as dysphagia and hoarseness and general symptoms such as fatigue and weight loss. Pathological examination revealed the presence of papillary carcinoma in the mass with presence of focus of papillary carcinoma in the thyroid bed in both cases. Periosseous invasion of the hyoid bone and involvement of the submandibular lymph nodes were observed in the second patient. The patients underwent total thyroidectomy with lymph node dissection followed by radioactive iodine therapy and are currently on thyroxin replacement. PMID- 18402126 TI - Incidental carcinoid tumors of the appendix: do they affect presentation or prognosis? AB - Carcinoid tumors are the most common neoplasm of the vermiform appendix, with most found incidentally after surgery for appendicitis. To determine in patients presenting with appendicitis if incidental carcinoid tumors affected presentation or prognosis, we reviewed our experience. Of 1350 patients receiving an appendectomy from 1992 to 2000 for appendicitis, 20 were found to have appendiceal carcinoid tumors. Comparing tumor patients to 251 consecutive patients whom underwent appendectomy without tumors, we found patients with tumors were older at presentation (41 +/- 4 versus 30 +/- 1 years, P = 0.005) and had initial lower white blood cell (WBC) counts (10.8 +/- 0.8 vs. 14.2 +/- 0.3, P = 0.007). There was equivalent survival between patients with carcinoid and without. Therefore, these data suggest that patients with carcinoid tumors are older and have lower WBC counts at presentation with appendicitis, although outcome was not affected by their presence. PMID- 18402127 TI - A rare and unusual case of bilateral, axillary, metachronous apocrine carcinoma. AB - Apocrine carcinoma is a rare sweat gland neoplasm with very few cases reported in the published literature. We report a case of primary axillary apocrine carcinoma with later recurrences in both axillae. A 55-year-old man was clinically diagnosed with hydradenitis suppurativa in the right axilla, and after excision of lesion, histology showed metastatic adenocarcinoma of probable breast origin. However, no primary focus was found after extensive work-up except for metastatic lymph nodes in the ipsilateral axilla treated with axillary clearance. After 4 years, the patient developed metastatic lymph nodes in the contralateral axilla and had surgery. He had a further recurrence in the right axilla and was treated with surgery and radiotherapy. He continues to do well at this time, with no evidence of local or distant metastases. PMID- 18402128 TI - Effect of Internet use on patient's surgical outcomes. AB - The Internet is an increasingly popular source of health care information, while 85% of physicians have experienced a patient bringing Internet information to a visit. Many physicians seem to be neutral toward inappropriate or incorrect information gathered by patients. To explore the impact of Internet use on surgical outcome and patients' overall satisfaction with the repair of inguinal hernia, patients presenting for hernia consult were given three preselected Internet sites and told to read up on the topic before the first surgical visit. 79.4% of the Internet group reported a very good to excellent postoperative level of satisfaction compared to 45.5% of the control group. When asked whether their expectations were met throughout the entire process, 74% of the Internet group reported "very much so" compared to 30% of the control group. The data encourage us to recommend that physicians review Internet health-related sites and make directed site referrals to their patients. PMID- 18402129 TI - Influence of synthetic mesh on ilioinguinal nerve motor conduction and chronic groin pain after inguinal herniorrhaphy: a prospective randomized clinical study. AB - Chronic postherniorrhaphy pain is a significant complication that can ruin a patient's quality of life. Our study aimed to assess the mesh-nerve interactions and the possible effects of this interaction on nerve morphology and function. Ilioinguinal nerve motor transmission studies using electromyelogram (EMG) were performed before the herniorrhaphy procedure, and Lichtenstein (n=50) or Shouldice (n=50) herniorrhaphies were used for repair. At the end of the first year after surgery, we found no significant differences on ilioinguinal nerve motor conduction studies between the mesh and Shouldice groups. We found a significant correlation between EMG results and inguinal pain. Our results indicates that nerve graft contact does not influence nerve motor conduction. The etiology of postherniorrhaphy chronic pain may be caused by nerve injury resulting in dissection or compression of the nerves. PMID- 18402130 TI - Solitary fibrous tumor of the pleura. AB - Solitary fibrous tumor of the pleura is a rare clinical entity. Approximately 600 such cases have been reported in the literature. The purpose of this study was to review the surgical outcome of this disease. This is a retrospective review of seven cases of solitary fibrous tumor of the pleura. There were five women and two men. Their ages ranged from 24 to 65 years. Symptoms included cough, shortness of breath, and chest pain. Diagnostic studies included chest X-ray, computed tomography (CT) scan, bronchoscopy, and CT-guided needle biopsy. All the tumors were pedunculated. The sizes ranged from 5.0 to 11.0 cm. All were resected by thoracotomy. The largest of these tumors weighed 68 g. All patients remain disease free. Follow-up ranged from 2 to 20 years. This is a rare disease. Complete resection offers the best chance for long-term care. PMID- 18402131 TI - Microbiological evaluation after gastrectomy and reconstruction for gastric cancer. AB - The purpose of this study was to evaluate microbial flora in the mucosa of reconstructed organs after gastrectomy for gastric cancer and improve postoperative quality of life by treating the flora. The number of aerobes was significantly higher in the gastric remnant in the proximal gastrectomy-jejunal pouch interposition group (n=8) than the distal gastrectomy-Billroth II reconstruction (G-BII) group (n=2) or the pylorus-preserving gastrectomy (PPG) group (n=8). The mean number and positive rate of anaerobes tended to be higher in jejunal pouch reconstruction groups. No Helicobacter pylori were detected in any specimens after the G-BII and jejunal pouch reconstruction. However, the gastric remnant and duodenum in the distal gastrectomy-Billroth I reconstruction group (n=5; positive rate of 80% and 20%, respectively) and the PPG group (positive rate of 63% and 25%, respectively) showed H. pylori. We concluded that more anaerobes tended to grow in the mucosa of reconstructed organs after jejunal pouch reconstruction than other procedures. Some patients after jejunal pouch reconstruction worried about their halitosis. Therefore, elimination of anaerobes may relieve it and improve postoperative quality of life. PMID- 18402132 TI - Cyst wall resection and ablation by hand-assisted laparoscopic surgery combined with argon plasma coagulator for huge hepatic cysts. AB - Six cases of deroofing of giant liver cysts by hand-assisted laparoscopic surgery (HALS) and safe and definitive cyst wall ablation with argon plasma coagulation (APC), which was verified by pathological examination of the resected cyst wall, were shown. PMID- 18402133 TI - Spatial modeling of human risk of exposure to vector-borne pathogens based on epidemiological versus arthropod vector data. AB - Understanding spatial patterns of human risk of exposure to arthropod vectors and their associated pathogens is critical for targeting limited prevention, surveillance, and control resources (e.g., spatial targeting of vaccination, drug administration, or education campaigns; use of sentinel sites to monitor vector abundance; and identifying areas for most effective use of pesticides). Vector borne disease risk can, in many cases, be modeled with high predictive accuracy by using geographic information system approaches because abundances of vectors and pathogen reservoirs often are associated with environmental factors. Spatial risk models for human exposure to vector-borne pathogens, which ideally should have high accuracy for predicting areas of elevated risk without overestimating risk coverage, can be constructed based on epidemiological data or abundance of vectors or infected vectors. We use five bacterial or viral vector-borne diseases occurring in the United States and with pathogen transmission by fleas (plague), ticks (Lyme disease and tularemia), or mosquitoes (dengue and West Nile virus disease) to 1) examine how spatial risk of human exposure to vector-borne pathogens typically is presented to the public health community and public and 2) evaluate the utility of basing spatial risk models on epidemiological data relative to data for arthropod vectors or infected vectors. Recommended future directions for vector-borne disease risk modeling include development of subcounty level spatial risk models combining epidemiological and vector data and the use of simulation or analytical models to assess critical vector abundance thresholds required for enzootic pathogen maintenance. PMID- 18402134 TI - Review of external morphology of Chirorhynchobiidae (Acari: Sarcoptoidea) with description of a new species. AB - Two species of Chirorhynchobiidae, Chirorhynchobia glossophaga sp. n. and Chirorhynchobia matsoni Yunker, 1970 are described and redescribed, respectively. The male of C. glossophaga and the tritonymph of C. matsoni are the first to be described in the family. The mites are ectoparasites living on the wing membrane of phyllostomid bats Glossophaga soricina (Pallas) from Peru and Anoura geoffroyi Gray from several localities. Females of the new species differ from those of C. matsoni by the shorter idiosoma (length 439-450 versus 497-562 in C. matsoni), by the dorsal position of setae 4a (terminal in C. matsoni), and by the narrower posterior projections of the propodonotal shield (width 6 versus 17). Although the morphology of Chirorhynchobiidae is strongly regressive, homologies of the leg and idiosomal setae in these mites with those of other Astigmata are established. The combination of absence of famulus epsilon and solenidia sigmaI II and presence of solenidia omega1I-II and omega3I in these mites is shared with the Rhyncoptidae-Sarcoptidae clade, suggesting a possible relationship with those groups. PMID- 18402135 TI - Identification of male specimens of the Culex pipiens complex (Diptera: Culicidae) in the hybrid zone using morphology and molecular techniques. AB - The identification of the members of the Culex pipiens L. complex in arbovirus surveillance programs relies heavily on the use of morphology. In this work, we studied Cx. pipiens complex male mosquitoes collected from nine different locations, from northern, southern, and the hybrid zone sites in North America; Cairo, Egypt; and Nairobi, Kenya. Specimens were identified using DV/D ratio and also using amplification of the acetylcholinesterase (ACE.2) gene by both conventional and real-time polymerase chain reaction, and examination of the CQ11 locus. Consistent with previous findings, the morphological and molecular identifications did not always agree, particularly in regions of extensive introgression. There was an increased frequency of hybrid forms in late summer and early fall in Champaign Co., IL, that is north of the previously described Cx. pipiens complex hybrid zone. This represents an expansion of the North American hybrid zone. The biological and epidemiological relevance of the high degree of introgression and the late season increase in the proportion of intermediate forms is discussed. PMID- 18402136 TI - Geographic variation in adult survival and reproductive tactics of the mosquito Aedes albopictus. AB - Climate differences across latitude can result in seasonal constraints and selection on life history characters. Since Aedes albopictus (Skuse) invaded North America in the mid-1980s, it has spread across a range of approximately 14 degrees latitude and populations in the north experience complete adult mortality due to cold winter temperatures that are absent in the south. Life table experiments were conducted to test for differences in the adult survival and reproductive schedules of Ae. albopictus females from two populations from the northern (Bloomington, IN [BL] and Manassas, VA [VA]; approximately 39 degrees N) and southern (Tampa, FL and Fort Myers, FL; approximately 27-28 degrees N) extremes of the species distribution in North America Regardless of population origin, age-specific hazard rate increased with reproductive output and decreased with number of bloodmeals. Larger females took fewer bloodmeals, and they had greater hazard rates than did smaller females. There were no consistent differences between northern versus southern populations in resource allocation between reproduction and maintenance, reproduction over time, and reproductive investment among offspring, suggesting that latitudinal variation in climate is probably not a main selective factor impinging on adult mortality and reproductive schedules. One possible effect of climate on geographic differences in life history was detected. BL had lower survivorship, lower lifetime reproductive output, and lower adult reproductive rate than did all other populations. This result may be an indirect result of lower egg survivorship due to the severity of winter in BL compared with other populations, including VA at approximately the same latitude. Such a scenario may make the BL population more prone to extinction, irregularly recolonized from more favorable sites, and thus more susceptible to founder effects, genetic drift, and inbreeding, resulting in lower mean values of fitness-related traits. PMID- 18402137 TI - Aggregation in the tick Ixodes ricinus (Acari: Ixodidae): use and reuse of questing vantage points. AB - Ongoing work in oak woods in Killarney National Park in southwestern Ireland is focusing on the factors influencing the fine-scale aggregated distribution of Ixodes ricinus L. (Acari: Ixodidae) on the ground. The extent of reuse of stems of vegetation as questing points by adult ticks was determined by paint-marking stems on which ticks were found, counting and removing these ticks, and subsequently reexamining the same stems for ticks on two further occasions. Overall, an estimated 2,967 stems in 123 separate rush plants (Juncus effusus L.) were examined. Statistical analysis of the data demonstrated a highly significant reoccupancy by ticks of stems previously and recently used. Furthermore, it is shown that the extent of stem reuse by ticks is significantly and positively correlated with the numbers of ticks originally observed on those stems. Although other factors may be involved in generating clumping of ticks, the results are compatible with the proposition that aggregation of I. ricinus on the ground is pheromone-mediated. The findings are discussed in relation to what is known about the powers of lateral movement of I. ricinus on the ground and the possible implications for the performance of tick traps. PMID- 18402138 TI - Mitochondrial and ribosomal internal transcribed spacer 1 diversity of Cimex lectularius (Hemiptera: Cimicidae). AB - Understanding genetic variation among populations of medically significant pest insects is important in studying insecticide resistance and insect dispersal. The bed bug, Cimex lectularius L. (Hemiptera: Cimicidae), is widespread hematophagus insect pest around the world, including North America, and it has recently been identified as an emerging resurgent pest. To date, no studies have been conducted on genetic variation of this species. For this study, 136 adult bed bugs representing 22 sampled populations from nine U.S. states, Canada, and Australia were subjected to genetic analysis using polymerase chain reaction (PCR) to amplify and sequence a region of the mitochondrial DNA (mtDNA) 16S rRNA gene and a portion of the nuclear rRNA internal transcribed spacer (ITS) 1 region. For the 397-bp 16S marker, a 12 nucleotide sites in total were polymorphic, and 19 unique haplotypes were observed. Heterozygosity was observed within many of the sampled populations for the mtDNA marker. This suggests that bed bug populations did not undergo a genetic bottleneck as one would expect from insecticide control during the 1940s and 1950s, but instead, that populations may have been maintained on other hosts such as birds and bats. In contrast to the high amount of heterozygosity observed with the mitochondrial DNA marker, no genetic variation in the 589-bp nuclear rRNA marker was observed. This suggests increased gene flow of previously isolated bed bug populations in the United States, and given the absence of barriers to gene flow, the spread of insecticide resistance may be rapid. PMID- 18402139 TI - A primer-introduced restriction analysis-polymerase chain reaction method to detect knockdown resistance mutations in Anopheles gambiae. AB - In the major malaria vector Anopheles gambiae Giles, two point mutations at the voltage-gated sodium channel have been associated with knockdown resistance (kdr) to DDT and pyrethroid insecticides. Simple allele-specific polymerase chain reaction (PCR) assays to detect these single-nucleotide polymorphisms are prone to lack of specificity and therefore alternative techniques have been proposed. However, these may not be easily implemented in many laboratories from malaria endemic regions. Here, we describe a primer-introduced restriction analysis (PIRA)-PCR method to detect kdr mutations in An. gambiae. This method unambiguously identified all six genotypes for the kdr locus in a sample of 113 field-collected mosquitoes for which kdr genotypes had been confirmed by DNA sequencing. Co-occurrence of both kdr alleles was found in sites from Equatorial Guinea and Gabon and the L1014F mutation was detected in M-form individuals from Angola. The PIRA-PCR proved to be a reliable, robust, and simpler alternative for the detection of kdr mutations in this malaria vector. PMID- 18402140 TI - Monooxygenase levels and knockdown resistance (kdr) allele frequencies in Anopheles gambiae and Anopheles arabiensis in Kenya. AB - Pyrethroid-treated bed-nets and indoor spray are important components of malaria control strategies in Kenya. Information on resistance to pyrethroid insecticides in Anopheles gambiae and An. arabiensis populations is essential to the selection of appropriate insecticides and the management of insecticide resistance. Monooxygenase activity and knockdown resistance (kdr) allele frequency are biochemical and molecular indicators of mosquito resistance to pyrethroids. This study determined baseline information on monooxygenase activity and kdr allele frequency in anopheline mosquitoes in the western region, the Great Rift Valley central region, and the coastal region of Kenya. In total, 1,990 field-collected individuals, representing 12 An. gambiae and 22 An. arabiensis populations were analyzed. We found significant among-population variation in monooxygenase activity in An. gambiae and An. arabiensis and substantial variability among individuals within populations. Nine of 12 An. gambiae populations exhibited significantly higher average monooxygenase activity than the susceptible Kisumu reference strain. The kdr alleles (L1014S) were detected in three An. gambiae populations, and one An. arabiensis population in western Kenya, but not in the Rift Valley-central region and the coastal Kenya region. All genotypes with the kdr alleles were heterozygous, and the conservative estimation of kdr allele frequency was below 1% in these four populations. Information on monooxygenase activity and kdr allele frequency reported in this study provided baseline data for monitoring insecticide resistance changes in Kenya during the era when large scale insecticide-treated bed-net and indoor residual spray campaigns were being implemented. PMID- 18402141 TI - Effects of physiological conditioning on behavioral avoidance by using a single age group of Aedes aegypti exposed to deltamethrin and DDT. AB - The behavioral and physiological responses of 6-d-old Aedes aegypti (L.) adult females exposed to deltamethrin and DDT were characterized using a free-choice excito-repellency test system. Excluding varying pretest age and carbohydrate availability as possible confounders, insecticide contact (measuring irritancy) and noncontact (measuring repellency) behavioral assays were conducted on two nonbloodfed groups, either unmated or mated (nulliparous), and two blood-fed groups, either parous or newly full-engorged mosquitoes. The degree of escape response to deltamethrin and DDT varied according to the physiological conditioning. Escape rates from contact and noncontact chambers with deltamethrin were more conspicuous in nonbloodfed groups compared with mosquitoes previously bloodfed. There were no significant differences in escape responses between unmated and nulliparous test populations. With DDT, a more pronounced escape response was observed in unmated compared with other physiological conditions. More moderate escape response was seen in nulliparous mosquitoes, and the least was observed in full bloodfed test individuals, regardless of test compound. Ae. aegypti, regardless of pretest conditioning, was completely susceptible to deltamethrin, whereas showing high resistance to DDT. Despite profound differences in resistance, there was no significant difference in avoidance response between chemicals and mosquito conditioning. Moreover, pre- and postbloodmeals were found to influence assay outcome and thus to have relevance on the interpretation of susceptibility and excito-repellency assays. PMID- 18402142 TI - Relationship between kdr mutation and resistance to pyrethroid and DDT insecticides in natural populations of Anopheles gambiae. AB - The spread of insecticide resistance genes in Anopheles gambiae Giles sensu stricto threatens to compromise vector-based malaria control programs. Two mutations at the same locus in the voltage-gated sodium channel gene are known to confer knockdown resistance (kdr) to pyrethroids and DDT. Kdr-e involves a leucine-serine substitution, and it was until recently thought to be restricted to East Africa, whereas kdr-w, which involves a leucine-phenylalanine substitution, is associated with resistance in West Africa. In this study, we analyze the frequency and relationship between the kdr genotypes and resistance to type I and type II pyrethroids and DDT by using WHO test kits in both the Forest-M and S molecular forms of An. gambiae in Cameroon. Both kdr-w and kdr-e polymorphisms were found in sympatric An. gambiae, and in many cases in the same mosquito. Kdr-e and kdr-w were detected in both forms, but they were predominant in the S form. Both kdr-e and kdr-w were closely associated with resistance to DDT and weakly associated with resistance to type II pyrethroids. Kdr-w conferred greater resistance to permethrin than kdr-e. We also describe a modified diagnostic designed to detect both resistant alleles simultaneously. PMID- 18402143 TI - Characterization of rickettsial infection in Amblyomma americanum (Acari: Ixodidae) by quantitative real-time polymerase chain reaction. AB - Several species of the spotted fever group Rickettsia (SFGR), with considerable variation in vertebrate host pathogenicity, are present in ticks in the United States. In this study, quantitative real-time PCR (qPCR) was used to characterize the growth and the distribution of Rickettsia amblyommii in selected tissues (salivary glands, gut, and ovaries) of naturally infected Amblyomma americanum (L.) (Acari: Ixodidae), during bloodmeal acquisition and throughout vertical transmission to eggs and postembryonic life cycle stages (larvae and nymphs). R. amblyommii was identified in the samples at ratios of < or = 1 rickettsiae per tick cell. Significant variability in the ratio of rickettsial to tick gene copy numbers between the tissues was identified; however, no single tissue was consistently observed to have the greatest rickettsial burden throughout the feeding event. Furthermore, the ratio of rickettsial to tick gene copy numbers did not significantly differ between eggs, immature ticks, and feeding events. This is the first study to use qPCR to enumerate rickettsial growth and distribution in the tick host during bloodmeal acquisition. Deciphering SFGR tissue distribution and transmission mechanisms is necessary for the development of novel approaches to control tick-borne rickettsial diseases. PMID- 18402144 TI - Host-seeking heights, host-seeking activity patterns, and West Nile virus infection rates for members of the Culex pipiens complex at different habitat types within the hybrid zone, Shelby County, TN, 2002 (Diptera: Culicidae). AB - Host-seeking heights, host-seeking activity patterns, and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) infection rates were assessed for members of the Culex pipiens complex from July to December 2002, by using chicken-baited can traps (CT) at four ecologically different sites in Shelby County, TN. Host seeking height was assessed by CT placed at elevations of 3.1, 4.6, and 7.6 m during one 24-h period per month. Host-seeking activity was assessed by paired CT placed at an elevation of 4.6 m. Can traps were sampled at one 10-h daytime interval and at seven 2-h intervals during the evening, night, and morning. Cx. pipiens complex mosquitoes accounted for 87.1% of collected mosquitoes. Culex (Melanoconion) erraticus (Dyar & Knab) accounted for 11.9% of specimens. The average number of Cx. pipiens complex mosquitoes collected per 24-h CT period from July to September was lowest at a rural middle income site (1.7), intermediate at an urban middle income site (11.3), and highest at an urban low income site (47.4). Can traps at the forested site failed to collect Cx. pipiens complex mosquitoes. From July to September at urban sites, Culex pipiens pipiens L. was the rarest of the three complex members accounting for 11.1-25.6% of specimens. At the rural site, Culex pipiens quinquefasciatus Say was the rarest member of the complex. Cx. p. pipiens was not collected after September. Mean abundance of Cx. pipiens complex mosquitoes was higher in traps at 7.6 m than in traps at 4.6 m. Abundances at 3.1 m were intermediate and not significantly different from abundances at the other heights. Initiation of host-seeking activity was associated with the end of civil twilight and activity occurred over an extended nighttime period lasting 8-10 h. All 11 WNV-positive mosquitoes were Cx. pipiens complex mosquitoes collected from urban sites in traps placed at elevations of 4.6 and 7.6 m. Infection rates were marginally nonsignificant by height. Infection rates, host-seeking heights, and activity patterns were not significantly different among members of the Cx. pipiens complex. PMID- 18402145 TI - Infection and co-infection rates of Anaplasma phagocytophilum variants, Babesia spp., Borrelia burgdorferi, and the rickettsial endosymbiont in Ixodes scapularis (Acari: Ixodidae) from sites in Indiana, Maine, Pennsylvania, and Wisconsin. AB - In total, 394 questing adult blacklegged ticks, Ixodes scapularis Say (Acari: Ixodidae), collected at four sites were analyzed by polymerase chain reaction (PCR) for five microbial species: Anaplasma phagocytophilum, Babesia microti, Babesia odocoilei, Borrelia burgdorferi, and the rickettsial I. scapularis endosymbiont. Identities of genetic variants of A. phagocytophilum were determined by sequencing a portion of the 16S DNA. In 55% of infected ticks (193/351), a single agent was detected. In 45% (158/351), two or more agents were detected; 37% harbored two agents and 8% harbored three agents. One male tick, collected from Ft. McCoy, WI, harbored all four microbial genera The highest rates of co-infection were by the Ixodes endosymbiont and B. burgdorferi (95/351). Two species of Babesia co-occurred within a single tick population in Wells National Estuarine Research Reserve, Wells, ME, whereas only B. odocoilei was found in other tick populations. Only A. phagocytophilum human anaplasmosis variant was detected in questing ticks from Tippecanoe River State Park, IN; from Wells; and Ft. McCoy, whereas a single infected tick from Presque Isle, PA, was infected by AP-Variant 1. Partially engorged ticks from deer in Tippecanoe River State Park were all infected with AP-Variant 1. Frequency of infections with each agent varied among populations. Rates and types of co-infections were not significantly different from random except for the Ixodes endosymbiont and B. burgdorferi in male ticks, which co-occurred less frequently than expected. Thus, I. scapularis hosts an array of pathogenic and symbiotic agents and potential evidence of interactions among microbial species was observed. PMID- 18402146 TI - Relative contribution of monooxygenase and esterase to pyrethroid resistance in Triatoma infestans (Hemiptera: Reduviidae) from Argentina and Bolivia. AB - Recently, high resistance to pyrethroid insecticides has been associated with ineffective field treatments against Triatoma infestans (Klug) (Hemiptera: Reduviidae) in northern Argentina. Samples were collected from two areas in Argentina (Salta and La Rioja) and one are in Bolivia (Yacuiba), and they were subjected to toxicological and biochemical assays. All populations were resistant to deltamethrin, but they showed different profiles to nonpyrethroid insecticides. The Salta population showed high resistance ratios (RRs) to deltamethrin and only slight differences in the susceptibility to fenitrothion and fipronil compared with the reference strain. Otherwise, the La Rioja population showed a lower RR to deltamethrin and no resistance to fenitrothion or fipronil. Finally, the Yacuiba population had high a RR to deltamethrin, but it was susceptibility to fenitrothion and fipronil. In several cases, deltamethrin resistant populations had higher susceptibility to bendiocarb than the reference strain. Measured activity of P450 monooxygenase in individual insects (based on ethoxycoumarine-O-deethylase), tended to be higher in the deltamethrin-resistant populations, but the differences were not statistically significant. Activity of specific esterases determined by the hydrolysis of 7-coumaryl permethrate demonstrated an increase in the percentage of insects with higher esterase activity in the Salta and La Rioja populations. Unexpectedly, the Yacuiba population showed lower pyrethroid esterase activity than the reference strain. The different pyrethroid resistance patterns found in T. infestans from three geographical regions within Argentina and in Bolivia suggests that enzyme-based pyrethroid resistance in this species has multiple origins. Nevertheless, because nerve insensitivity (related to the presence of the kdr gene) is also an important mechanism related to pyrethroid resistance, further studies on the kdr gene should be carried to clarify the relative contribution of each pyrethroid associated mechanism in deltamethrin-resistant populations of T. PMID- 18402147 TI - Age-dependent survival of the dengue vector Aedes aegypti (Diptera: Culicidae) demonstrated by simultaneous release-recapture of different age cohorts. AB - The assumption that mosquito survival remains constant and that it is independent of age was tested with free-ranging Aedes aegypti (L.) (Diptera: Culicidae) by using a "synthetic cohort" mark-release-recapture method. Mosquito age cohorts (1 2, 3-4, and 19-20 d) were released simultaneously in homes in rural Thai villages during dry and rainy seasons (2002). Significant age-dependent effects were detected during the dry and rainy seasons. More young than older mosquitoes were recaptured (1-4 versus 5-20 d). The best predictive fit for recaptures was obtained using a quadratic function of age. Our results provide the first field based evidence for age-dependent Ae. aegypti mortality in which the death rate increases with advancing age and highlight the need for research on ecological and epidemiological aspects of this process. PMID- 18402148 TI - Investigation of ecological and environmental determinants for the presence of questing Ixodes ricinus (Acari: Ixodidae) on Gower, South Wales. AB - The spatial heterogeneity of questing Ixodes ricinus (L.) (Acari: Ixodidae) within endemic areas in Great Britain is well established. Their presence is acutely responsive to blood host availability and their ability to maintain water balance, which are in turn governed by a variety of ecological and environmental factors. This article details the findings of a 3-yr study on the Gower peninsula, south Wales, which investigated the contribution of such factors (both ground- and geographic information systems [GIS] -derived) for predicting the presence of questing I. ricinus (Q(P)), at a local scale. Statistically significant univariate associations were found between Q(P) and calcareous/ neutral grassland and heathland habitats, particularly those grazed by livestock, and various factors that intuitively promote tick survival. For example, topographical features, such as certain aspects, that reduce exposure to cold northerly winds and the hot midday sun, favored Q(P). Similarly, positive associations were found with substrata composed of less permeable soil types and less permeable superficial/bedrock geologies that promote a moist microhabitat and reduce the likelihood of desiccation. Q(P) was also higher in areas of high soil moisture. This study highlighted a number of GIS-derived data sets that could be applied in the development of local and national predictive maps for I. ricinus in Great Britain. An understanding of the influence of these factors on questing I. ricinus can aid targeted tick control programs and help to educate the public, and those occupationally exposed, in understanding likely I. ricinus prolific areas within an I. ricinus endemic region. PMID- 18402149 TI - Bloodmeal-induced differential gene expression in the disease vector Culex nigripalpus (Diptera: Culicidae). AB - We have characterized gene expression changes in the midgut tissue of Culex nigripalpus Theobold (Diptera: Culicidae) females, after they ingest a bloodmeal, by differential display of RNA isolated from pre- and postfeeding females. Seventy-two cDNA fragments exhibiting reproducible differences were observed. Of these, 22 showed an increase in gene expression and 50 showed a decrease in gene expression as a result of blood feeding. Eleven cDNA fragments exhibiting increased expression after a bloodmeal were cloned and sequenced. Seven of these were novel transcripts, and they did not match any sequences in GenBank. Three clones were similar to conserved proteins of unknown function from Aedes aegypti (L.) and Anopheles gambiae Giles. Here, we present the expression data on the first cDNA clones isolated from Cx. nigripalpus midgut tissue, including their molecular characterization, and we discuss their possible involvement in blood feeding-associated processes and disease transmission. PMID- 18402150 TI - Pupal dimensions as predictors of adult size in fitness studies of Aedes aegypti (Diptera: Culicidae). AB - Adult body size is a central life history character in mosquito fitness studies. I evaluated the predictive values of pupal cephalothorax length, cephalothorax width, and wet weight for adult size (wing length) of male and female Aedes aegypti (L.) (Diptera: Culicidae). Cephalothorax length was the most consistent and accurate predictor of adult size. Width of the cephalothorax and wet weight were more variable, and they significantly decreased shortly before adult emergence. I propose that cephalothorax length could be used as a proxy for adult size to test how physical and biological factors such as resource-limited environments and competition affect mosquito fitness with the advantage that the specimen does not need to be killed. PMID- 18402151 TI - Taxes well paid. PMID- 18402152 TI - Effect of Alzheimer's training on multicultural personal care aides. AB - This article reports on the first 2 years of an ongoing project that examined the efficacy of a 10-hour dementia training provided to entry-level personal care aide (PCA) trainees from the Hispanic, White, African American, and Asian communities in New York City. Participants were enrolled in a 90-hour PCA training program offered by the New York City Department for the Aging and were either recipients of public assistance, displaced employees from September 11, or recent immigrants to the United States from China. Classes were conducted in Spanish, English, and Mandarin/ Cantonese. An 11-item Knowledge of Alzheimer's Disease instrument was developed for the purposes of this project and administered before and after the dementia training and at 3 months following graduation. All groups, regardless of language, showed a significant increase in knowledge of Alzheimer's disease at the conclusion of the training and retention of this knowledge at 3 months follow-up. Age was strongly correlated with an increase in knowledge, while gender and education were not. PMID- 18402153 TI - Medicare part D: legal analysis. PMID- 18402154 TI - Infinite possibility: clowning with elderly people. AB - Over the past 30 years, there has been aplentitude of research into the health benefits of humor and laughter for healthy, sick, or depressed adults and children as well as for senior citizens. Medical research supports our human instinct that people who smile and laugh are happy, whereas those who are inexpressive are usually not happy. Research shows that humor stimulus results in mirth, which elicits a primarily emotional response with psychological efects, and laughter, which elicits a physical response with physiological effects. The many physiological benefits of laughter in older adults have been clearly demonstrated. Yet much of the medical research is based on experiments using funny videos and cartoons for humor sessions. I argue that "clowning around" with elderly people brings greater benefits than laughter alone. These benefits are clearly evident, though they may not be scientifically measurable: When the game is rooted in the patient's own imagination, thereby giving agency to a powerless individual it is many times more personal and transformative. In this article, I focus on my experiences with older adults while working with Clowns Without Borders and Risaterapia as well as on my own relationship with my grandfather. I provide a framework for why humanitarian clowning and the principles behind it can be incredibly well suited for working with the elderly. PMID- 18402155 TI - Medical care for the homeless elderly. AB - This is a case study of two elderly, frail women in New York City who were recently rendered homeless. One woman had a massive tumor on her occipital scalp; the other was in renal failure. The obstacles and complexities of providing care to those with double jeopardy--being elderly and homeless--are described. There are enormous difficulties for placement into safe, supportive housing once people become homeless. The process is expensive and labor intensive. This can be complicated by the existence of mental illness. A New York agency that works with mentally ill homeless people is described. There are systemic obstacles as well: One woman loses her Medicaid when she moves from one state to another to be closer to her family. Another, 82 years old, is told to get a job so that she could qualify for Medicare. There are numerous contradictions and unnecessary costs in a fragmented health care system to which the obvious solution is a national single-payer system of care. PMID- 18402156 TI - Confident living program for senior adults experiencing vision and hearing loss. AB - Many people experience both vision and hearing losses as they age. The Confident Living Program was developed by Helen Keller National Center to address the unique psychosocial and educational needs of older adults living with dual sensory impairments. PMID- 18402157 TI - [Session: on the subject: "Healthcare response to terrorism armed conflict excepted". Introduction]. PMID- 18402158 TI - [Treatment of radiation burns with surgery and cell therapy. A report of two cases]. AB - Treatment of severe radiation burns remains a difficult challenge. Conventional surgical treatment (excision, skin grafting, skin or muscle flaps) often fails to prevent unpredictable and uncontrolled extension of the necrotic process. We report two clinical cases in which surgery was combined with mesenchymal stem cell (MSC) therapy. Clinical outcome was good and there was no recurrence of radiation inflammatory waves observed in the first patient after one year. This novel multi-disciplinary therapeutic approach, combining physical techniques, modern plastic surgery and cell therapy should improve the medical management of severe localized radiation burns. PMID- 18402159 TI - [Chemical terrorism and the healthcare response]. AB - Many chemical products can be used for terrorist purposes, as they have the potential for seriously intoxicating many people and creating a risk of secondary contamination. Despite precautions taken by the authorities with respect to emergency responses and equipment for protection, decontamination and treatment, victim management remains difficult. Theoretical and practical training is compulsory. Fundamental research, notably in enzymatic and biomolecular engineering, has identified very promising proteolytic agents that may be used for protection and treatment of those exposed to organophosphorus nerve agents and insecticides. PMID- 18402160 TI - [Healthcare response to bioterrorism]. AB - Bioterrorists seek to use highly virulent pathogens to cause social and economic disruption. Historical agents such as anthrax, Yersinia pestis and botulinum toxin can easily be produced, whereas others such as hemorragic fever virus require much more sophisticated infrastructure and government-scale resources. In the near future, with the spread of new biotechnologies, bioterrorists may produce new transgenic pathogens. Epidemiological surveillance and diagnostic capacities should thus be more reactive. Real-time epidemiological surveillance together with a network of high-security laboratories would enable us to cope effectively with new infectious diseases that emerge naturally or through human manipulation. Clinical and research skills should be merged in infection-control centers, bringing a new dynamic to microbiology. PMID- 18402161 TI - [From alert to laboratory: a coherent network designed to deal with to naturally occurring infectious disease outbreaks and bioterrorism]. AB - Public health prevention requires early detection of disease outbreaks, whether naturally occurring or due to bioterrorism. Permanent surveillance and a network of laboratories are the two main pillars of effective outbreak management. Coordination of information, training, and procedures are under the responsibility of the French public health watch institute and the scientific advisory board for the Biotox-Piratox laboratory network. Protective capacities against bioterrorism are improving but efforts must continue. PMID- 18402162 TI - [Psychological impacts of terrorism on victims and the general population]. AB - Terrorism is a major public health concern. The impact of violence against the civilian population is reinforced by the media reporting. Thus, terrorism has a psychological impact not only on its direct victims but also on the population as a whole. More research is needed on how to manage these consequences. PMID- 18402163 TI - [Persistent Bartonella infection: epidemiological and clinical implications]. AB - Bartonella are Gram-negative hemotropic bacteria that infect a wide range of mammals. At least 14 Bartonella species or subspecies have been reported to be pathogenic for humans and animals. Wild and domestic animals represent a large reservoir. Reservoir species usually display chronic bacteremia. This explains some aspects of the epidemiology of these infections, and especially vector-borne transmission. The molecular mechanisms of persistent infection have clinical consequences both for occasional hosts and for human and animal reservoirs. An increasing number of clinical cases are being described in reservoir species that were previously considered to remain asymptomatic. PMID- 18402164 TI - [Immunization against varicella and zoster]. AB - Two vaccines against varicella-zoster virus are available in France. These live attenuated vaccines are derived from the Oka strain used in Japan since 1974. They are indicated for healthy subjects from 12 months of age, at a dose of one injection until 12 years of age, and two injections 4-8 weeks apart for older children and adults. Seroconversion occurs in 95% of cases and the antibodies persist beyond 5 years. Clinical efficacy is about 85% against all forms of varicella and nearly 100% against severe forms. Post-exposure vaccination within 3 days may also prevent the disease. A universal immunization program against varicella was implemented in the USA in 1995. Now, with vaccine coverage at about 80%, the incidence of the disease has been reduced by 85%, with the largest decrease in 1- to 4-year-olds. Tolerability is generally good, with only mild reactions at the injection site and moderate fever The length of protection is not yet known. A two-dose schedule seems advisable to avoid breakthrough varicella, which occurs in 4% of vaccinees each year. Insufficient coverage is expected to lead to later disease onset, with more severe cases in adolescents and adults. Universal immunization could also increase the incidence of zoster. These problems indeed seem to be emerging in the United States. France has adopted restrictive guidelines on VZV vaccination, but they are expected to be revised when the combined MMR-V vaccine becomes available. Zoster vaccine, prepared with the same strain but at a higher concentration, has moderate efficacy on zoster and on post-zoster neuralgia in patients over 70. This vaccine is not yet recommended in France, because the length of protection is not known and there is a potential risk of delaying the occurrence of zoster and, thus, of increasing the risk of post zoster neuralgia. PMID- 18402165 TI - [Computed tomography and cranial paleoanthropology]. AB - Since its invention in 1972, computed tomography (C.T.) has significantly evolved. With the advent of multi-slice detectors (500 times more sensitive than conventional radiography) and high-powered computer programs, medical applications have also improved. CT is now contributing to paleoanthropological research. Its non-destructive nature is the biggest advantage for studying fossil skulls. The second advantage is the possibility of image analysis, storage, and transmission. Potential disadvantages include the possible loss of files and the need to keep up with rapid technological advances. Our experience since the late 1970s, and a recent PhD thesis, led us to describe routine applications of this method. The main contributions of CT to cranial paleoanthropology are five-fold: -Numerical anatomy with rapid acquisition and high spatial resolution (helicoidal and multidetector CT) offering digital storage and stereolithography (3D printing). --Numerical biometry (2D and 3D) can be used to create "normograms" such as the 3D craniofacial reference model used in maxillofacial surgery. - Numerical analysis offers thorough characterization of the specimen and its state of conservation and/or restoration. --From "surrealism" to virtual imaging, anatomical structures can be reconstructed, providing access to hidden or dangerous zones. --The time dimension (4D imaging) confers movement and the possibility for endoscopic simulation and internal navigation (see Iconography). New technical developments will focus on data processing and networking. It remains our duty to deal respectfully with human fossils. PMID- 18402166 TI - [Gene expression profiling in colon cancer]. AB - Identification of new prognostic factors for colon cancer with no lymph node involvement may improve the selection of patients for adjuvant chemotherapy. The aim of this study was to assess the possibility of using gene expression profiling for this purpose. Fifty patients operated on for stage II colon cancer were included. Twenty-five of these patients relapsed, while the other 25 remained disease-free for at least 5 years. MRNA was extracted from fresh-frozen biopsies and hybridized to the Affymetrix GeneChip HGU133A. One thousand six hundred random splits of the 50 patients into a training set and a validation set were considered. For each split, a prognostic combination was derived from the training set (selection of the 30 genes most differentially expressed between patients who recurred and those who did not, by linear discriminant analysis), and its prognostic performance was assessed with the validation set. On average, accuracy was 76%, sensitivity 85%, and specificity 68%. A total of 6,124 genes were included in at least one of the 1,600 predictive combinations, and 55 genes were included in more than 100 combinations. This study supports the possibility of predicting the prognosis of non-metastatic colon cancer by tumor gene expression profiling. It also shows the highly variable gene composition of predictive combinations. PMID- 18402167 TI - [Epidemiology and risk factors of inflammatory bowel diseases]. AB - Inflammatory bowel diseases (IBD) are a public health problem in industrialized countries, where 1 in 1000 people are affected Most patients are young adults. The incidence of IBD has increased considerably in western countries since the second world war but is beginning to level off. On the other hand, the incidence is still rising in low-incidence areas such as Eastern Europe, Asia and developing countries. Differences in incidence rates across age, time, and geographic areas suggest that environmental factors are involved in IBD, but only cigarette smoking and appendectomy have consistently been identified as risk factors. An important role of genetic factors in IBD was first suggested by epidemiological studies showing familial aggregation of IBD and by twin studies. In 2001, the first CD susceptibility gene, NOD2/CARD15 on chromosome 16, was characterized. Other susceptibility genes have since been located. Their identification should help to understand the complex interaction between the environment and the intestinal immune system. PMID- 18402168 TI - [Treatment of chronic inflammatory bowel diseases]. AB - The same drugs are used to treat hemorrhagic rectocolitis (HRC) and Crohn's disease (CD), although the aims are very different. Aminosalicylates are highly beneficial in HRC but virtually ineffective in CD, a disease in which immunosuppressants are more useful. Aminosalicylates exert their antiinflammatory effect directly on the lesions. Various oral and rectal preparations have been developed in order to deliver the active molecule to the intestinal target segment. These drugs are now known to act by stimulating the nuclear receptor PPAR-gamma, and this knowledge should help with the development of new agents. Chronic aminosalicylate treatment appears to diminish the risk of malignant transformation. Systemic steroids are still the mainstay of treatment for exacerbations of HRC and CD, yielding remissions in 60 to 90% of cases. In contrast, systemic steroids should not be used for maintenance therapy. Budesonide is a preparation that selectively releases steroids in the ileocolonic region, thereby reducing systemic adverse effects. Immunosuppressants such as azathioprine and 6-mercaptopurine, and also methotrexate in some cases, are used for maintenance therapy of steroid-dependent and highly recurrent forms. These drugs stabilize the disease in about half the patients who receive them. Treatment typically lasts several years and necessitates regular monitoring, especially of hematological status. Cyclosporine is used intravenously in severe HRC. Infliximab (RemicadeR), a chimeric monoclonal antibody targeting TNF is effective in acute forms and as maintenance therapy for CD. It was also recently shown to be effective in HRC. Infliximab is indicated in steroid-resistant forms and contraindicated in patients with latent systemic infections (tuberculosis, hepatitis B) and heart failure. Artificial nutrition is now only used in children with acute forms, in order to avoid the need for steroids. Probiotics might have a place in maintenance treatment of HRC. Surgical treatment of HRC consists of colectomy or, more radically, ileoprotectomy followed by ileoanal anastomosis with resection: however, the likely benefits and potential complications (especially nocturnal diarrhea) must be carefully weighed up. Surgical treatment of CD consists of resecting the worst-affected segments. As available medical and surgical treatments can only control these diseases, without curing them, patient management must be planned on a long-term basis. Control of exacerbations is judged on the basis of clinical parameters and biological markers of inflammation, rather than on lesion healing. The choice of maintenance therapy depends on the nature of the disease (HRC or CD) and its progressive nature. Surgery is reserved for patients with complicated and drug-resistant forms. PMID- 18402169 TI - [Crohn's disease: current surgical treatment]. AB - Surgical treatment of Crohn's disease (CD) is purely symptomatic. In addition, medical therapy always precedes surgery and almost always continues afterwards. The indications for surgical treatment are failure of medical treatment and progressive complications such as chronic bowel obstruction and occlusion, internal abscesses and fistulae, and recurrences. Between 70 and 80% of patients with CD will undergo surgical treatment at some time, mainly reflecting the very high frequency of recurrences (50% at 10 years). Laparoscopic surgery has many advantages in this setting, except for urgent interventions. In the last 30 years, segmental small-bowel resection has followed precise technical intestine saving rules. Widening enteroplasty ("stricturoplasty") must replace resection for patients with staged CD and multiple foci, and must be used for patients who have already had mutilating resections for multiple recurrences, in order to avoid the "short bowel" syndrome. In severe acute colitis, early surgery is indicated if short-term resuscitation fails: the operation is always subtotal colectomy with double stomy of the ileum and of the sigmoid colon. In chronic and scalable colorectal attacks, the choice of technique depends on the location and severity of the lesions. Conserving the distal colon is justified if the lesions are moderate: this saves natural transit for a time, which is important for young adults. At least half of these conservative treatments eventually fail. Progressive pancoloproctitis complicated by anoperineal lesions that compromise continence is usually treated by total coloproctectomy with final ileostomy. Sphincter conservation by ileoanal anastomosis is only possible in rare patients with colorectal CD. PMID- 18402170 TI - A picture to be painted. PMID- 18402171 TI - Re: Ethnic minority patients. PMID- 18402172 TI - Continuous professional development (CPD) is an obligation for all health professionals. PMID- 18402173 TI - Noise annoys. PMID- 18402174 TI - Sounds lively! Choirs: introducing singing into healthcare. PMID- 18402175 TI - Time being: difficulties in integrating arts in health. PMID- 18402176 TI - Challenging stigma around mental illness and promoting social inclusion using the performing arts. AB - This article outlines the rationale, evidence base, method and qualitative evaluation of a project that uses the performing arts to challenge the stigma surrounding mental illness and promote social inclusion of people with mental health problems. The partnership project has run for three years with students of Reigate Sixth Form College and staff and users of voluntary and statutory mental health services in East Surrey. Collation of three years of evaluation data showed it as a successful approach to: positively influence students' attitudes, knowledge and empathy around mental health issues; and positively affect mental health service clients' mood and their feelings of achievement, confidence and inclusion. Key elements of the project's success and sustainability are summarised. PMID- 18402177 TI - 'Singing for the brain': reflections on the human capacity for music arising from a pilot study of group singing with Alzheimer's patients. AB - The paper reports the activities of a pilot study of group singing for people with Alzheimer's and their carers, and presents data arising from the study. AIMS: The project aimed to answer the following questions: Would people with Alzheimer's be able to participate in group singing? Would progress be identifiable in participants' singing? Would responses to the activity be positive? Would participating carers find the activity worthwhile? Additionally, the following question was posed: Within the supportive environment of group singing, might people with Alzheimer's be able to participate in a song that they had to learn? METHODS: Songs were selected that would be familiar to participants aged over 40 years, and one original song was introduced by the group leader following a successful session. The group leader did not know which of the participants were patients with Alzheimer's and which were carers. Participants sat in a circle in a resonant hall. Sessions were filmed and a separate audio recording made and subsequently analysed, and questionnaires were completed by carers after sessions. RESULTS: Nine hours of video recording, three hours of audio recording and three collated questionnaires (one for each session) were collected. Confidence grew over the three sessions, with a traceable development in the alertness of many of the people with Alzheimer's. It proved possible to divide the group so as to sing two songs simultaneously, and also so as to perform three- and four-part rounds. It proved possible to teach an unknown song. Carers generally felt that the sessions were of value. CONCLUSIONS: People with Alzheimer's are able to participate in group singing and some longer-term benefits are perceived by their carers. In a group activity such as Singing for the Brain it is difficult to be certain how the overall effect arises from the interaction of individuals. The data assembled is difficult to submit to clinical testing, relying as it does on the judgement of participants regarding reported recall of the content of sessions on the part of people with Alzheimer's. Further research questions are raised by the success of this project. PMID- 18402178 TI - Does partnered dance promote health? The case of tango Argentino. AB - AIMS: Cultural activities, including music, singing and dance, have often been conceptualised as enhancing well-being as well as promoting mental and physical health. This paper focuses on partnered dance in the context of 'cultures of fitness'. Its purpose was to investigate the dancers of tango Argentino, and to explore potential health benefits in this group. METHODS: Participants (N=110) completed a self-developed inventory, which was, in part, based on interviews and observations as a participant observer. The inventory addresses educational, musical, and socio-economic background, motivation for and investment in dancing tango, and other leisure activities. RESULTS: Dancers are characterized by high level education and socio-economic status. Motivation appears to be predominantly driven by both hedonistic and social factors, accounting altogether for nearly 60% of the variance. The majority of individuals started dancing tango only in their 30s. Physical investment in terms of time and money indicate tango dancing as a highly important activity that seems to involve substantial opportunity of moderate physical exercise, social interaction and emotional reward. In consequence, tango dance was identified as the primary leisure activity. CONCLUSIONS: Tango dancers were identified as highly educated individuals, who are dedicated to the practice in many respects, including physical and emotional. They draw from similar motivations as compared to participants of other musical activities such as singing and listening to music. Notably, aspects of physical fitness are more prominent in this group. The intensity with which tango is practised seems to depend to some degree on individual lifestyles. Taken together, the results suggest a high degree of personal involvement in tango dance as a primary leisure activity. Implications of partnered dance for social, emotional, and physical well-being and health promotion demand further research. PMID- 18402179 TI - The impact of art, design and environment in mental healthcare: a systematic review of the literature. PMID- 18402180 TI - Human intestinal parasites. PMID- 18402181 TI - Perceptions of women about menstrual regulation services: qualitative interviews from selected urban areas of Dhaka. AB - Menstrual regulation (MR) programmes were introduced in Bangladesh in 1974 to reduce morbidity and mortality due to unsafe abortions. About 468,000 MR procedures are performed annually, and its potential is not fully used. To develop MR programmes, the voices of women could add important aspects to its acceptability. This qualitative interview study aimed to explore and describe the perceptions about MR in a sample of women from Dhaka, Bangladesh. The most prominent perception was that, despite the moral dilemma inherent in terminating pregnancies, MR was highly valued as a solution in problematic life situations. However, unprofessional attitudes and misconduct among MR providers were revealed, and there was also a lack of knowledge and openness in families. To improve the quality of MR services, professional ethics needs to be highlighted in training and supervision of providers. To improve the acceptability of MR, education on the benefits of MR has to be made available to the whole population. PMID- 18402182 TI - Prevalence of intestinal parasites in young Quichua children in the highlands of rural Ecuador. AB - The prevalence of intestinal parasites in young Quichua children was assessed in 20 rural communities in the highlands of Ecuador in August 2005. The caregivers of 293 children aged 12-60 months were interviewed about the status of child health, household socioeconomic and environmental factors, and water-use practices and were requested to collect a faecal sample from the study child. Two hundred three (69.3%) of the 293 children provided faecal samples that were tested for parasites. The overall prevalences of infection for specific agents were Entamoeba histolytica or dispar 57.1%, Ascaris lumbricoides 35.5%, Entamoeba coli 34.0%, Giardia intestinalis (lamblia) 21.1%, Hymenolepis nana 11.3%, Cryptosporidium parvum 8.9%, Chilomastix mesnili 1.7%, Hymenolepis diminuta 1.0%, Strongyloides stercoralis 0.7%, and Trichuris trichiura 0.5%. The prevalence of parasites increased with age. Water storage, water treatment, consistent latrine use, and participation in a community-based clean water project were not strongly associated with the prevalence of intestinal parasites, although having dirt floors was a risk factor for infection with E. histolytica or dispar and G. intestinalis. PMID- 18402183 TI - Prevalence, haemolytic and haemagglutination activities and antibiotic susceptibility profiles of Campylobacter spp. isolated from human diarrhoeal stools in Vhembe District, South Africa. AB - Campylobacter species are increasingly being recognized as agents of gastroenteritis worldwide. However, data on the pathogenic characteristics of the organism isolated in rural communities in South Africa are lacking. In this study, the prevalence of Campylobacter spp. from diarrhoeal stools, haemolytic and haemagglutinating activities of the isolates, and antibiotic susceptibility profiles, including minimum inhibitory concentration (MIC) patterns to different antibiotics, were determined using the standard microbiological techniques. Campylobacter spp. were isolated from individuals of all age-groups; however, the infection rate was higher among individuals aged less than two years (30.4%). Of 115 Campylobacter strains isolated, polymerase chain reaction (PCR) analysis indicated that 98 (85%) were C. jejuni, while 17 (15%) were C. coli. Seventy-one (62%) of the strains showed haemolysis on human blood, and 80% agglutinated human blood, whereas 22.6% were beta-lactamase-positive. Resistance to antimicrobials, such as erythromycin, ciprofloxacin, vancomycin, and fusidic acid, was high. Increased resistance to macrolide and quinolone antibiotics poses major risks for treatment failure. Haemolytic and haemagglutinating activities may be useful in preliminary characterization of pathogenic strains in settings where Campylobacter-associated infections are common. PMID- 18402184 TI - Acute otitis media during the first two years of life in a rural community in Bangladesh: a prospective cohort study. AB - The study investigated the burden of acute otitis media (AOM) during the first two years of life in a cohort of 252 newborns in rural Bangladesh using data collected on occurrences of AOM. Trained community health workers (CHWs) conducted household surveillance and picked up cases of AOM using the study algorithm. The incidence rate was 0.9 episodes per child-year observed. Forty-six percent (n=115) of the 252 subjects developed AOM: 36% (n=91) during the first year of life and 10% (n=24) during the second year of life (p<0.001). The age specific incidence rates of AOM varied; peaks occurred in the 6-12-month age group and the lowest in the first three months of life. In total, 20% (n=49) of the study subjects had single, 26% (n=66) recurrent, and 54% (n=137) no episode of AOM. Perforation with discharge developed in 85% (n=322) of 375 episodes. The duration of discharge from the ears was < or =6 weeks in 95% of the episodes, but in 5% of the episodes, discharge from the ears continued for >6 weeks. The incidence of AOM was higher in the monsoon season compared to the summer season (p<0.003). The study documented AOM as an important cause of morbidity among rural children up to two years of age in Bangladesh and should be addressed with strategies to overcome the burden of disease. PMID- 18402185 TI - Prevalence of Helicobacter pylori in children and their family members in a district in Turkey. AB - The aim of this study was to determine the prevalence of Helicobacter pylori among children and their family members and to evaluate some epidemiologic characteristics. The study included 275 children, aged 1-15 year(s), suffering from different gastrointestinal complaints. Blood serology and stool antigen testing were used for the diagnosis of infection due to H. pylori. Sixty-five (23.6%) of the 275 children were positive for H. pylori, and this positivity had a significantly increasing correlation with age (p<0.001). H. pylori-associated infection was observed among 45 (69.2%) and 17 (8%) mothers in the H. pylori infected and non-infected groups respectively (p<0.0001). Most children and their families infected with H. pylori were living in an urban area. The findings suggest that infection due to H. pylori is a problem for this district area, and all children having any gastrointestinal complaints should be examined whether H. pylori was prevalent among their family members. PMID- 18402186 TI - Prevalence, pathogenesis, antibiotic susceptibility profiles, and in-vitro activity of selected medicinal plants against Aeromonas isolates from stool samples of patients in the Venda region of South Africa. AB - The prevalence, pathogenic indices, such as haemolytic and haemagglutinating activities, antibiograms, and in-vitro activities of local medicinal plants against Aeromonas isolates in Vhembe district of Limpopo province, South Africa, were studied using standard microbiological methods. In total, 309 diarrhoeic stool samples were collected from patients attending five health centres in the region during December 2004-May 2005. Aeromonas species were identified using the API 20E system. The haemagglutinating and haemolytic activities of isolates on human, sheep, pig and chicken red blood cells were investigated. Antibiotic susceptibility profiles of the isolates to several antibiotics and in-vitro activity of local medicinal plants were also ascertained using previously reported schemes. Results showed that 104 (33.6%) of the 309 samples were positive for Aeromonas species, of which 89 (85.6%) were Aeromonas hydrophila, 12 (11.5%) A. sobria, and three (2.9%) A. caviae. All strains of A. hydrophila and A. caviae produced haemolysis on sheep blood, while eight of the 12 A. sobria strains were haemolytic on sheep blood. The haemolytic activities of the isolates were variable on other red blood cells tested. High level of resistance was observed to amoxicillin and ampicillin, followed by cefuroxime (79%), chloramphenicol (74%), and erythromycin (65%). The carbapenems were the most active drugs with only 7% resistance to meropenem and 11% to imipenem. About 12% of the isolates were resistant to ciprofloxacin. The extracts of three of seven medicinal plants tested showed inhibitory activity against all Aeromonas isolates; these included acetone and hexane extracts of Pterocarpus angolensis, Syzygium cordatum, and Zornia milneana. The results suggest a high prevalence of Aeromonas species in the region. The isolates demonstrated multiple resistant profiles to different antibiotics tested. Some local medicinal plants were inhibitory to Aeromonas isolates, indicating a potential role in the management of Aeromonas-related infections. Structural elucidation of the active components may pave the way for the discovery of candidate templates for eventual drug design. Most isolates possessed important virulence characteristics based on their haemolytic and haemagglutinating ability. However, the genetic characterization of the isolates will further confirm their pathogenicity and the origin of multiple antibiotic resistance. PMID- 18402187 TI - Development and management of a geographic information system for health research in a developing-country setting: a case study from Bangladesh. AB - In the last decade, geographic information systems (GIS) have become accessible to researchers in developing countries, yet guidance remains sparse for developing a GIS. Drawing on experience in developing a GIS for a large community trial in rural Bangladesh, six stages for constructing, maintaining, and using a GIS for health research purposes were outlined. The system contains 0.25 million landmarks, including 150,000 houses, in an area of 435 sq km with over 650,000 people. Assuming access to reasonably accurate paper boundary maps of the intended working area and the absence of pre-existing digital local-area maps, the six stages are: to (a) digitize and update existing paper maps, (b) join the digitized maps into a large-area map, (c) reference this large-area map to a geographic coordinate system, (d) insert location landmarks of interest, (e) maintain the GIS, and (f) link it to other research databases. These basic steps can produce a household-level, updated, scaleable GIS that can both enhance field efficiency and support epidemiologic analyses of demographic patterns, diseases, and health outcomes. PMID- 18402188 TI - A review of changing episode definitions and their effects on estimates of diarrhoeal morbidity. AB - This paper describes how the methodology used for measuring diarrhoeal morbidity has changed over time and assesses how differences in episode definition have affected estimates of diarrhoeal morbidity among children aged less than five years. The episode definition used in 73 studies included in three previously published literature reviews was identified. In earlier work, a method was developed that adjusts morbidity estimates to take account of differences in episode definition. This adjustment method was applied to the studies identified in these three literature reviews. Episode definitions were better documented and were more consistent in studies published after 1980. Adjusting morbidity estimates to account for definitional differences did not substantially alter the reviews' conclusions. Diarrhoeal surveillance has steadily improved since 1980, with methodology becoming more consistent between studies and better documented. Although episode definitions have changed over time, the morbidity estimates derived in the three reviews appear robust to these changes. PMID- 18402189 TI - Sociodemographic characteristics of tobacco consumers in a rural area of Bangladesh. AB - Bangladesh typifies many developing countries experiencing an increasing trend in tobacco consumption. However, little is known about the general pattern of tobacco consumption and about population groups who are more prone to tobacco consumption. This paper aimed at generating knowledge on tobacco consumption, especially emphasizing the identification of sociodemographic groups who are more prone to tobacco consumption vis-a-vis tobacco-related health consequences in a remote rural area in Bangladesh. Information on the tobacco consumption status of 6,618 individuals (52.1% males, 47.9% females), aged over 15 years, was collected in 1994. Both univariate and multivariate analyses were done. Individuals were categorized as consumers if they consumed tobacco in any form at all, i.e. smoke or chew. The independent variables included various characteristics of individuals and households. Overall, 43.4% of the study subjects consumed tobacco. Males were 9.38 times more likely to consume tobacco than their female counterparts. Individuals with no education were 3.62 times more likely to consume tobacco than those who had completed six or more years of schooling, and the poor were almost twice as likely to consume tobacco than the rich. Tobacco consumption in both smoke and chewing form has been a part of household consumption in Bangladesh from time immemorial. Only aggressive anti-tobacco programmes on various fronts may salvage the vulnerable groups from the menace of tobacco consumption in Bangladesh. PMID- 18402190 TI - Successful treatment of rotavirus-induced diarrhoea in suckling mice with egg yolk immunoglobulin. AB - The role of specific immunoglobulins at mucosal sites in imparting protection against disease, such as rotavirus-associated diarrhoea, is well-established. Oral immunoglobulin therapy with egg yolk-derived anti-rotavirus immunoglobulins has previously been shown to achieve moderate therapeutic effect in diarrhoea due to rotavirus in a clinical trial. Here, data on the therapeutic potential of the same immunoglobulin preparation in an infant mouse model of rotavirus-induced diarrhoea is presented. The use of an animal model has allowed therapy to be evaluated with higher doses of immunoglobulins and has suggested that an improved therapeutic effect can be achieved by increasing the dose in the clinical setting. PMID- 18402191 TI - Adults' perceived prevalence of enteric fever predicts laboratory-validated incidence of typhoid fever in children. AB - This study was undertaken to develop a model to predict the incidence of typhoid in children based on adults' perception of prevalence of enteric fever in the wider community. Typhoid cases among children, aged 5-15 years, from epidemic regions in five Asian countries were confirmed with a positive Salmonella Typhi culture of the blood sample. Estimates of the prevalence of enteric fever were obtained from random samples of adults in the same study sites. Regression models were used for establishing the prediction equation. The percentages of enteric fever reported by adults and cases of typhoid incidence per 100,000, detected through blood culture were 4.7 and 24.18 for Viet Nam, 3.8 and 29.20 for China, 26.3 and 180.33 for Indonesia, 66.0 and 454.15 for India, and 52.7 and 407.18 for Pakistan respectively. An established prediction equation was: incidence of typhoid (1/100,000= -2.6946 + 7.2296 x reported prevalence of enteric fever (%) (F=31.7, p<0.01; R2=0.992). Using adults' perception of prevalence of disease as the basis for estimating its incidence in children provides a cost-effective behavioural epidemiologic method to facilitate prevention and control of the disease. PMID- 18402192 TI - Delivery practices of traditional birth attendants in Dhaka slums, Bangladesh. AB - This paper describes associations among delivery-location, training of birth attendants, birthing practices, and early postpartum morbidity in women in slum areas of Dhaka, Bangladesh. During November 1993-May 1995, data on delivery location, training of birth attendants, birthing practices, delivery-related complications, and postpartum morbidity were collected through interviews with 1,506 women, 489 home-based birth attendants, and audits in 20 facilities where the women from this study gave birth. Associations among maternal characteristics, birth practices, delivery-location, and early postpartum morbidity were specifically explored. Self-reported postpartum morbidity was associated with maternal characteristics, delivery-related complications, and some birthing practices. Dais with more experience were more likely to use potentially-harmful birthing practices which increased the risk of postpartum morbidity among women with births at home. Postpartum morbidity did not differ by birth-location. Safe motherhood programmes must develop effective strategies to discourage potentially-harmful home-based delivery practices demonstrated to contribute to morbidity. PMID- 18402193 TI - Body mass index of male youths aged 18-20 years of the Han nationality living in different regions of China. AB - The study was conducted to assess the nutritional status and levels of body mass index (BMI, kg/m2) and to evaluate the geographical distribution of male youths of the Han nationality in China. In total, 60,773 male youths, aged 18-20 years, of the Han nationality, were categorized into underweight, normal-weight, overweight, and obesity according to the international adult BMI cut-offs. Different levels of nutritional status and BMI of male youths of the Han nationality were compared among different areas. The mean BMI for the whole country was 20.6 in urban areas and 20.0 in rural areas. BMI increased from 20.1 among 18-year old youths to 20.5 among 20-year old youths. The prevalence of underweight among the male youths was 21.6%, while the prevalence of overweight and obesity were 4.6% and 0.6% respectively. For urban youths, the prevalence of underweight, overweight, and obesity were 21.0%, 6.8%, and 1.1% respectively, while these were, respectively, 21.9%, 3.3%, and 0.3% for rural youths. The nutritional status of the male youths in North-China was at the highest level (21.1) among the six areas, and the prevalence of underweight, overweight, and obesity were 14.3%, 9.1%, and 1.4% respectively. The highest prevalence of underweight was 29.8% in the North-West region, and the lowest prevalence of overweight was 2.2% in the South-Middle region, while the lowest prevalence of obesity was 0.2% in the South-West region. The nutritional status of the male youths was significantly different among different areas. Underweight was still prevalent in all male youth groups. Nonetheless, overweight was more prevalent among urban youths than among rural youths and was more prevalent in the North region than in the South region. PMID- 18402194 TI - Newborn Aides: an innovative approach in sick newborn care at a district-level special care unit. AB - A Sick Newborn Care Unit (SNCU), established in a district hospital in India, substantially reduced the neonatal mortality rate in the district; it, however, suffered from a dearth of trained nurses. Local girls with 10-12 years of school education underwent structured and hands-on training for six months, followed by a six-month internship at the SNCU and were assigned to it as stipendiary 'Newborn Aides'. Based on the results of formal examinations, internal on-the-job assessment and interview of doctors, nurses, and parents and their technical skills and motivation were rated very high. Although the incremental cost of training is small, the cost of sustaining them, i.e. stipend and replacing attrition, needs to be addressed. Trained Newborn Aides may substantially alleviate human-resource constraint for SNCUs and Sick Newborn Stabilization units in smaller peripheral hospitals for care of sick newborns at an affordable cost. PMID- 18402195 TI - Preoperative evaluation of lung resection candidates. Preface. PMID- 18402196 TI - Spirometry: predicting risk and outcome. AB - Predicted postoperative FEV1 is certainly the most widely used parameter in preoperative risk stratification [54] and the measure recommend by BTS and ACCP functional guidelines as a first step in the screening of patients for lung resection surgery. Nevertheless, recent evidences have demonstrated that ppoFEV1 is not a reliable predictor of postoperative cardiopulmonary complications in patients with preoperative impaired pulmonary function. This may be because of the fact that the resection of a portion of lung in patients with obstructive disease determines only a minimal loss, or even an improvement, in overall respiratory function and exercise tolerance. This lung volume reduction effect takes place very early, since the first postoperative days, balancing what ever negative physiologic effects a thoracotomy and lung resection may entail. In addition to its poor predictive role in COPD patients, ppoFEV1 largely underestimate the actual loss in the very first days after operation, when most of the complications develop. The rationale to use a parameter which is poorly correlated with the pulmonary function at the moment the complications occur seems unwarranted. At the very best, ppoFEV1 appears a weak surrogate of the immediate postoperative FEV1. The FEV1 measured on the first postoperative day may be 30% less than predicted. Corrective equations have been published to correct this discrepancy with the aim to improve risk stratification. PMID- 18402197 TI - The role of diffusing capacity and exercise tests. AB - Beside pulmonary function tests, other measures of cardiopulmonary fitness have been shown to be useful for preoperative risk stratification. Reduced values of carbon monoxide lung diffusion capacity and its predicted postoperative value have been reported to be associated with postoperative complications and mortality. The most widely used low-technology exercise tests, which include 6 minutes of walking and stair climbing, have been reported to be valid predictors of complications. Patients with an impaired performance at these tests need to undergo a formal cardiopulmonary test with measurement of maximum oxygen consumption at cycle-ergometry or treadmill. Functional algorithms have been proposed that incorporate all these tests, with the aim to assist in defining the surgical risk. PMID- 18402198 TI - Assessing regional lung function. AB - The assessment of regional lung function to predict postoperative function is integral to preoperative evaluation of pulmonary resection candidates who have impaired lung function. The four validated ways are: anatomic calculation, split radionucleotide perfusion scanning with single photon emission CT (SPECT), quantitative CT scanning, and dynamic perfusion MRI. Estimations of predicted postoperative lung function based on anatomical calculations are the simplest, tend to overestimate postresectional functional loss, and should be performed in most operative candidates. Patients requiring further evaluation require one of the other techniques: SPECT, quantitative CT, or MRI. All effectively predict postoperative forced expiratory volume in 1 second (FEV1). Cost, local expertise and availability, and the need to predict postoperative VO maximum oxygen consumption (VO2 max) dictate the clinician's choice. PMID- 18402199 TI - Future trends in preoperative evaluation. AB - Preoperative evaluation before lung resection has been frequently addressed in modern medical literature. Actual or predicted pulmonary volumes are considered relevant to predict the risk of surgery. Nevertheless, ppoFEV1 underestimates the real functional loss in the immediate postoperative period when most of the complications occur. Not all patients, however, have comparable functional changes after lobectomy. Minimal impairment or even improvements have been demonstrated in COPD cases after lobectomy. Efforts should be directed to an accurate prediction of the immediate postoperative pulmonary volumes for a better evaluation of high-risk patients caused by respiratory impairment. Future developments are needed on the role of measuring preoperative DLCO and how to evaluate a patient's general cardiorespiratory status. Evidence underlines the relevance of routine evaluation of preoperative DLCO at rest or, better, during exercise for a thorough assessment of patient's capability to adapt to a stressful situation (Fig. 3). Only by improving knowledge about the general condition of the patient, can one assess the physiologic response to surgery. Widespread use of sophisticated or simple exercise tests and measurements or daily activity using motion detectors can identify high-risk patients with otherwise acceptable pulmonary volumes. Another suggested investigation issue is to develop different relevant outcome parameters, not only from the surgeon's point of view but also from the patient's perspective, such as postoperative QOL related variables or delayed outcomes. Finally, multidisciplinary investigation teams, including experts in mathematical modeling, are essential to improve the quality and validity of the developed models. Although knowledge about perioperative physiologic changes has increased, clinicians are still far from finding a way to put all this knowledge down and make it applicable for an individual patient. Multicentric cooperation and evaluation of large prospectively recorded databases are essential to develop evidence-based clinical guidelines on preoperative evaluation. PMID- 18402200 TI - Preoperative and postoperative pulmonary rehabilitation in lung cancer patients. AB - The benefits derived from comprehensive PR, when applied to patients who have lung cancer, should have significant impact on both survival and health status. Because PR is known to improve exercise capacity, it is reasonable to expect that this treatment modality may provide more patients with a potential cure. In addition, improvement in symptoms and quality of life can prove critically important when long-term survival is not an outcome that can be impacted on. Studies thus far support the value of this treatment modality in the global approach to patients who have lung cancer. Future well-designed clinical trials will need to corroborate these findings. We look forward to improving lung cancer outcomes with the widespread use of pulmonary rehabilitation. PMID- 18402201 TI - Preoperative cardiac evaluation of lung resection candidates. AB - Patients who have lung cancer typically have both pulmonary and cardiac disease as a result of cigarette smoking and are potentially at increased risk for perioperative cardiopulmonary complications. Knowledge of risk factors and a careful preoperative assessment will help the medical team stratify the patient's level of risk and employ measures to minimize surgical risk. The available literature specific to cardiac risk and lung cancer surgery is minimal, but the general principles of preoperative cardiac risk evaluation and perioperative management have been reviewed. When considering cardiac testing and interventions, the medical consultant must remember that surgery is the treatment of choice for non-small cell lung cancer and must avoid any excessive delay that might compromise the patient's chance of a surgical cure. PMID- 18402202 TI - Predicting the need for intensive care following lung resection. AB - On the basis of the evidence available, the authors would suggest a decision making algorithm to determine the need for ICU admission postoperatively similar to that shown in Fig. 1. First, patients should quit smoking at least 1 month and preferably 2 months before surgery. Those over the age of 70 years should receive elective ICU admission. Second, those at increased risk of general anesthesia, as judged by ASA and performance status scores and cardiovascular risk assessment, should be prebooked into the ICU in the postoperative period. A ppo FEV1 of less than 44% should warrant additional monitoring rather than mandate ICU admission. Pre-existing fibrotic lung disease mandates ICU admission. Third, perioperatively, protective (low tidal volume) ventilatory strategies should be applied during one lung ventilation. Patients undergoing one lung ventilation, and especially those undergoing extensive lymphatic dissection, should be monitored closely for signs of ALI in the first 5 days postoperatively. This, together with any indication of postoperative complications such as POP, BPF or empyema, should mandate immediate transfer to the ICU. PMID- 18402203 TI - Risks of neoadjuvant chemotherapy and radiation therapy. AB - Pulmonary resection for locally advanced NSCLC after induction chemotherapy or chemoradiotherapy can generally be performed with acceptable morbidity and mortality. Data from several phase II and a few randomized trials showed that patients should be carefully selected for surgery especially with respect to their age, performance status, and pulmonary and cardiovascular function tests. The presence of cardiovascular disease should be actively investigated and preexisting arrhythmias appropriately managed. Similarly, respiratory status should be evaluated and any reversible condition, such bronchial obstruction, infection, pulmonary embolism, and smoking, should be addressed. Surgical resection is technically more demanding; intraoperative blood loss should be minimized. Several studies demonstrated that pneumonectomy especially on the right side is associated with a significantly increased risk of postoperative morbidity and mortality especially in patients after induction chemoradiotherapy. Therefore, pneumonectomy should be performed very selectively and only when alternative procedures such as bronchial or vascular sleeve resection or both are technically not possible. Long-term follow-up reports of bimodality protocols without surgery versus bimodality followed by surgical resection are awaited. These results will heln to define standards of care and the role of surgery for patients with NSCLC stage III disease. Future decision-making will have to take into account treatment morbidity and mortality and parameters of organ-sparing surgery following induction. probably best represented by rates of pneumonectomy or rates of lobectomy in different patient groups. PMID- 18402204 TI - Long-term quality of life after lung resection. AB - Several studies have shown that physical domains of QOL deteriorate early after lung cancer surgery but improve to baseline by 6 months after surgery. Impaired cardiopulmonary function following pulmonary resection is an important predictor of immediate postoperative morbidity; meanwhile, an objective assessment of pulmonary function did not significantly contribute to QOL in LTLC survivors. Chronic pain is the most common complication after lung cancer surgery. Lung cancer survivors do not experience the same length of life and QOL as their age matched peers or other cancer survivors. Lung cancer patients are less well than survivors of other cancers because of the nature of the cancer and the smoking history. There is clearly a need to identify and intervene with subgroups of patients who are at an elevated risk of premature death and diminished QOL. PMID- 18402205 TI - Parenchymal-sparing resections: why, when, and how. AB - Although lobectomy should continue to be regarded as the procedure of choice for NSCLC, certain subsets of patients who have favorable characteristics may be treated appropriately with segmentectomy without adversely affecting oncologic outcome as long as an adequate assessment of intraoperative nodal status and surgical margin is performed. The use of anatomic segmentectomy may be particularly useful for small, peripheral tumors less than 2 cm in diameter located within anatomic segmental boundaries, as well as for elderly patients who have impaired cardiopulmonary function. Ground-glass opacities and lesions displaying bronchoalveolar histology also may be appropriate target lesions for segmentectomy because of their low metastatic potential. Prospective, randomized studies (such as the CALGB/Altorki trial and the brachytherapy mesh trial) will be necessary to delineate fully the utility of segmentectomy in patients who have NSCLC. PMID- 18402206 TI - The use of scoring systems in selecting patients for lung resection: work-up bias comes full-circle. AB - The construction of statistical models of perioperative risk and long-term postoperative survival is a useful activity. It facilitates fair, assessment of surgical outcomes and provides insight into the association between certain clinical features and outcome. It provides quantitative estimates of risk or long term survival. There are, however, a number of limitations to the use of such models in informing decisions concerning the selection of patients for lung resection. In essence, the limitations described in this article are those of work-up bias come full circle. Concerning the use of scoring systems in selecting patients for resection, one should remember the advice of the wise Gene Blackstone: caveat emptor. The findings of model-building exercises, if based on surgical databases, can only augment, and not replace, clinical judgment. When models suggest that certain patient groups do well, the prior selection of these patients should be borne in mind. When models of perioperative risk or long-term survival suggest that certain patient groups, despite being carefully selected by clinical teams, do badly, this information should be heeded. That said, moves to deny informed patients lung resection on the basis of estimates of risk or "poor" survival should be considered carefully. For example the British Thoracic Society Guidelines on Surgery for lung cancer state that mortality following resection should not be in excess of 4% for lobectomy. It is not exactly clear what is intended by publishing that statement. It represents some form of audit standard but clearly fails if one thinks in terms of the individual patient. A patient who has a curable cancer and who faces a life expectancy likely to be under 2 years without surgery might well accept a greater than 1 in 25 chance of perioperative death. If 25 patients in a room were facing that prospect, all 25 might reasonably hope to be among the 24 expected survivors and opt for surgery. PMID- 18402207 TI - The influence of preoperative risk stratification on fast-tracking patients after pulmonary resection. AB - Fast-tracking protocols or postoperative care computerized algorithms have been shown to reduce hospital length of stay and reduce costs; however, not all patients can be fast-tracked. Certain patient characteristics may put patients at increased risk to fail fast-tracking. Additionally some patients have multiple risk factors that have an additive effect that puts them at an even increased risk to fail fast-tracking, and more importantly, to significant morbidity. It is a mistake to force these protocols on all patients because it can lead to increased complications, readmissions, and low patient and family satisfaction. By carefully analyzing surgical results via accurate prospective databases, the types of patients who fail fast-tracking and the reasons they fail can be identified. Once these characteristics are pinpointed, specific changes to the postoperative algorithm can be implemented, and these alterations can lead to improved outcomes. The authors have shown that by using pain pumps instead of epidurals in elderly patients we can improve outcomes and still fast-track octogenarians with minimal morbidity and high-patient satisfaction. We have also shown that the increased use of physical therapy and respiratory treatments (important parts of the care of all patients after pulmonary resection, but a limited resource in most hospitals) may also lead to improved surgical results for those who have low FEV1% and DLco%. Further studies are needed. Although fast tracking protocols cannot be applied to all, the vast majority of patients who undergo elective pulmonary resection, even those at high risk, can undergo safe, efficient, and cost-saving care via preset postoperative algorithms. When the typical daily events are convened each morning and the planned date of discharge is frequently communicated with the patient and family before surgery and each day in the hospital, most patients can be safely fast-tracked with high satisfaction and outstanding results. PMID- 18402208 TI - [Evaluation of lipid digestion using the 13C-mixed triglyceride breath test in patients with chronic pancreatitis]. AB - BACKGROUND: Steatorrhea represents the indirect sign of lipid maldigestion in chronic pancreatitis and even when the measurement of fecal fat is considered as a gold standard for the diagnosis of steatorrhea, this test is not commonly used within clinical practice because of the inconvenience related to sample collection. Although the use of breath test using mixted tryglicerides was initally validated as an indirect alternative for the assessment of exocrine pancreas reserve, only recently has used this method as a surrogate for the measurement of fat in feces. AIM: To evaluate fat digestion by means of the breath test with 13C labelled mixed triglycerides in patients with chronic pancreatitis. MATERIAL AND METHODS: Patients with chronic pancreatitis underwent clinical and biochemical evaluation. The latter included serum amylase, lipase, betacarotenes; fecal fat analysis and breath test using 13C-mixed tryglicerides. Breath test results are expressed as the percentage of 13C recovered in the breath sample. RESULTS: Seventeen patients (age: 45 +/- 5 years) were included, of which 7 had steatorrhea (fecal fat greater than 7 g/day). In patients with steatorrhea, the percentage of recovered 13C from breath was significantly lower (6 +/- 4%) than in patients without it (25 +/- 5%). CONCLUSION: Results suggest the uselfuness of breath test with 13C-mixed tryglicerides as an alternative for the assessment of lipid digestion in patients with chronic pancreatitis. PMID- 18402209 TI - [Usefulness of peristalsis, flatulence and evacuation for predicting oral route tolerance in patients subjected to major abdominal surgery]. AB - OBJECTIVE: to evaluate the usefulness of bowel sounds, flatus and bowel movement presence to predict tolerance of oral intake in patients following major abdominal surgery. BACKGROUND DATA: nutrition is one of the most important factors in the management of postoperative care. The early oral intake has shown to contribute to a faster recovery. Traditionally the beginning of postoperative feeding after major abdominal surgery is delayed until bowel sounds, flatus and/or bowel movement are present although there is no enough medical evidence for their usefulness. METHODS: We studied 88 patients following major abdominal surgery. We registered the presence of bowel sounds, flatus and bowel movement each 24 hours in the postoperative period. We analized the relationship between the presence of these signs and the ability to tolerate oral intake. Predictive values, sensitivity, specificity and ROC curves were calculated. RESULTS: results shown that bowel sounds have an acCeptable sensibility but a very low specificity to predict the ability to tolerate oral intake. Unlike bowel sounds, bowel movements shown a low sensibility and a high specificity. Flatus turned out to have and intermediate sensitivity and specificity in the prediction of tolerance of oral feeding. CONCLUSIONS: in this study any of these signs were shown as a reliable indicator for beginning oral feeding because they have a moderate to low usefulness. PMID- 18402210 TI - [Obesity, symptoms of gastroesophageal reflux and endoscopic findings in a referral hospital]. AB - OBJECTIVE: To determine association between obesity, gastroesophageal reflux symptoms, hiatus hernia and erosive esophagitis. METHODS: Consecutive patients who underwent upper endoscopies at our center were studied. Before endoscopy, through a direct interview all subjects were asked to complete the Carlsson-Dent questionnaire. Weight and height of all patients were determined. Body mass index was calculated. At endoscopy it was looked for the presence of hiatus hernia and erosive esofagitis. We excluded patients with some of the following conditions: Pregnancy, ascitis, esophageal varices, esophageal, gastric or duodenal stenosis, and patient who required an emergency or therapeutic endoscopy. We compared prevalence of symptomatic GERD, hiatus hernia and erosive esophagitis between normal weight patients, overweight and obese. RESULTS: A total of 196 patients were included for analysis, 122 women and 74 men, mean age 52.5 years, 40.3% were overweight and 28.1% were obese. There were 124 patients (63.3%) with symptoms of GERD determined by the C-D Questionnaire obtained. Hiatus hernia was observed in 87 patients (44.4%), and erosive esophagitis in 69 patients (35.7%). Prevalence of GERD symptoms in obese or overweight patients were similar to those normal weight patients (66.4% and 56.5% respectively, p = 0.20) OR 1.52 (IC 95%, 0.82 2.82). Among obese or overweight hiatus hernia was observed in 47% compared to 38.7% of those normal-weight (p = 0.28) OR 1.40 (IC 95%, 0.76-2.59). Prevalence of erosive esofagitis was also similar in both groups 37.3% and 32.3% respectively (p = 0.52) OR 1.25 (IC 95%, 0.66-2.36). CONCLUSIONS: This study didn't show association between symptoms of GERD, hiatus hernia or erosive esofagitis and overweight or obesity. PMID- 18402211 TI - [Detection and treatment of subclinical hypothyroidism in patients with irritable bowel syndrome. Does it modify the use of health resources?]. AB - BACKGROUND: It is still unknown whether the treatment and control of conditions that coexist with irritable bowel syndrome (IBS) change the utilization of healthcare resources for this disease. OBJECTIVES: To determine whether the treatment of subclinical hypothyroidism in patients with IBS decreased the use of healthcare resources and work absenteeism. MATERIAL AND METHODS: Patients with IBS and subclinical hypothyroidism (cases) and patients with IBS and no hypothyroidism (controls) were diagnosed during a three-year period. Other conditions were ruled out by means ofa group of tests. Four years later, a survey on the associated medical events, the use of healthcare resources, and work absenteeism during the previous year was applied. Controls were age- and gender matched at a 2:1 control: case ratio. RESULTS: Fourteen cases and 28 controls were studied, all of them females, with a mean age of 46 years. The survey was conducted 4 years after the initial diagnosis in both groups. No significant differences were found between both groups in the following: the proportion of the patients who had received medical treatment for IBS (79% of cases vs. 82% of controls), the prescription of psychotropics (36% vs. 36%, respectively), the proportion seen in the emergency room due to gastrointestinal complaints (21% vs. 18%, respectively), job absenteeism (7% vs. 11%, respectively), and the proportion of patients who underwent abdominal surgery (14% vs. 11%, respectively). The mean number of visits to the doctor due to gastrointestinal complaints within the year prior to the interview was similar in both groups (3.1 for cases vs. 3.4 for controls). CONCLUSIONS: The detection and treatment of subclinical hypothyroidism do not modify the utilization of healthcare resources in IBS. PMID- 18402212 TI - [Complications in diagnostic and therapeutic endoscopic cholangiopancreatography. Prospective study]. AB - BACKGROUND: Studies of ERCP-related morbidity seldom include a sufficient patient follow-up. The complication rate is variable. AIM: To characterize and to evaluate the frequency of complications in patients after Diagnostic and Therapeutic ERCP. PATIENTS AND METHODS: All patients undergoing ERCP during a 2 year period were included in this prospective study. Complications were assessed at time of ERCP and by personal or telephone contact at 1st, 7, 14 and 30-days after the procedure. RESULTS: A total of 897 ERCPs were included in the analysis, of which 93.9% were therapeutic procedures. 640 were female with age of 49.68 (+/ 18.59) and 257 male with age 58.80 (+/-16.67). The 30-day complications rate was 3.19%; the procedure-related mortality rate was 0.11%. Mild hemorrhage occurred in 1.48% and moderate hemorrhage in 0.11%. Post-ERCP pancreatitis occurred in 0.79% and duodenal perforation in 0.45%. The 30-day procedure-no related mortality was 1.14%. CONCLUSION: This prospective study confirms the complications rate of ERCP including therapeutic procedures is low in our experience. PMID- 18402213 TI - [Bone marrow infiltration as an early sign of gastric cancer]. AB - The symptomatic infiltration to bone marrow by gastric carcinoma is an extremely infrequent condition even so as an early manifestation. Very few reports exist in the worldwide literature about it. In this paper we report a case of a 47 years woman who was attended in our Institute because of the presence of thrombocytopenia, anaemia, fever involuntary weight loss and linfadenopathy, and also dyspepsia. Through her diagnostic evaluation the diagnosis of an occult gastric cancer was established. The aim of this report is to present the clinical, radiological and pathological characteristics of this unusual presentation of gastric cancer; and also we present a brief literature review of the cases that had been reported and their clinical implication. PMID- 18402214 TI - [Autoimmune hepatitis and primary sclerosing cholangitis: dependent or independent association?]. AB - BACKGROUND AND OBJECTIVE: The diagnosis of an autoimmune liver disease is based on clinical, biochemical, immunological and histological criteria particular to each disease, such as autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) or primary sclerosing cholangitis (PSC). Hepatic overlap syndromes (OS) are defined by the presence of a well established autoimmune liver disease, primarily AIH plus two or more characteristics associated with another liver disease (PBC or PSC); however the association of HAI and PSC is considered rare in adult population, and only 43 cases have been reported in the literature. The aim of this study is to review and reappraise the characteristics of this rare association, and to discuss current concepts on OS. METHODS: We describe the clinical presentation, evolution, radiologic studies, histological characteristics and therapeutic options in a Mexican woman with OS (HAI-PSC). We also include an updated review of the literature. RESULTS: Overlap ofAIH and PSC has been described in a number of repqrts during the last decade, and is assumed to exist in a considerable part of mainly young patients with autoimmune liver diseases. Sequential appearance of AIH and PSC has been described in children, but also may be observed in adults. This association has been reported between 1.4%-8%, probably because differences in age of the study populations, range of autoantibodies taken into consideration, and degree of completeness of analyzed data. CONCLUSIONS: HAI-PSC is a rare disease, more common in children, and its outcome and evolution seem to be similar of AIH alone. Ursodesoxycholic acid in combination with an immunosuppressive regimen may be an adequate medical treatment for most patients with this association, and liver transplantation should be considered in late stage disease. PMID- 18402215 TI - [Total gastrectomy in the management of Menetrie's disease. Institutional experience and review of the literature]. AB - OBJECTIVE: Analyze retrospectively the clinical, histopathological and surgical characteristics of patients with Mindtrier's disease (MD) at our Institution. BACKGROUND DATA: Menetrier's disease (MD) includes hipertophy of gastric mucosa, hipo or hiperchloridria and severe hipoalbuminemia. In the literature, it has been underestimated its relationship with the development of gastric carcinoma. METHODS: We analyzed retrospectively the clinical trials of patients with MD treated surgically at our Institution during the last three decades (From 1970 through 2002). RESULTS: Three patients with clinical diagnosis of MD underwent total gastrectomy during the study period. All of them were corroborated histopathologically and in one patient carcinoma in situ was demonstrated in the surgical specimen. CONCLUSIONS: There is a very limited experience with surgical treatment of MD and it is limited to patients who have severe hipoalbuminemia or associated malignant disease. However accordig to our experience and literature review, surgical treatment should be considered early in the course of the disease for the relationship between MD and gastric adenocarcinoma. PMID- 18402216 TI - [Clinical images in gastroenterology. Pyogenic liver abscess with pleural complication and Papillon-Lefevre syndrome]. PMID- 18402217 TI - [Clinical images in gastroenterology. Acuphagia]. PMID- 18402218 TI - [AGA Institute technical review on acute pancreatitis]. PMID- 18402219 TI - [Clinical guidelines for the diagnosis and treatment of dyspepsia. Epidemiology, definition, classification]. PMID- 18402220 TI - [Clinical guidelines for the diagnosis and treatment of chronic ulcerative colitis (CUC). Diagnosis. Usefulness of clinical data and laboratory analysis]. PMID- 18402221 TI - [Clinical guidelines for the diagnosis and treatment of chronic ulcerative colitis (CUC). Diagnosis. Imaging]. PMID- 18402222 TI - [Clinical guidelines for the diagnosis and treatment of chronic ulcerative colitis (CUC). Medical treatment]. PMID- 18402223 TI - [Clinical guidelines for the diagnosis and treatment of chronic ulcerative colitis (CUC). Treatment of mild to moderate CUC. Remission induction and its maintenance]. PMID- 18402224 TI - [Clinical guidelines for the diagnosis and treatment of chronic ulcerative colitis (CUC). Treatment of severe CUC and followup approach for cancer risk assessment]. PMID- 18402225 TI - [Third Mexican Consensus on Helicobater pylori]. PMID- 18402226 TI - [III Mexican Consensus on Helicobacter pylori]. PMID- 18402227 TI - [On the applicability of the North American principlism]. AB - The four prima facie principles proposed for bioethics by Beauchamp and Childress (principlism) has been one of the most followed bioethical paradigms, gaining a prominent place not only in United States, but in many other parts of the world. This model has been criticized from different points of view, especially in the last fifteen years. One of the major objections to principlism is the difficulty of applying the system to real bioethical dilemmas. The essential problem of the system, apart from an absence of hierarchy among the four principles, is an absence of a clear framework of human goods (anthropologically speaking) which are necessary to protect. Consequently, in the face of an ethical dilemma two opposite solutions can be reached using one principle or another. The authors offer as a solution to these difficulties the specification and consideration of the principles and rules. However, in many cases, this is not sufficient. PMID- 18402228 TI - [The religious convictions in the argumentation bioethics. Two different secularists perspectives: Sadaba and Habermas-Rawls]. AB - This article analyses the position of two secularized theories on the role of religious beliefs in bioethical reasoning. The excluding laicism of Sadaba rejects the rationality of religious fact and extend a general suspicion about the bioethical reasoning of believer. Contrary, the open position of Habermas Rawls considers reasonable religions as one of the typical comprehensive views of liberal State, encourage secularized citizens to value his contributions and urge to secular and, then, neutral, State not to impose to all citizens a secularized cosmo-vision. Only the second perspective put the bases for a fruitful and calm dialogue in the bioethical area. PMID- 18402229 TI - [Bioethics of principles]. AB - Bioethics emerges about the tecnological problems of acting in human life. Emerges also the problem of the moral limits determination, because they seem exterior of this practice. The Bioethics of Principles, take his rationality of the teleological thinking, and the autonomism. These divergence manifest the epistemological fragility and the great difficulty of hmoraln thinking. This is evident in the determination of autonomy's principle, it has not the ethical content of Kant's propose. We need a new ethic rationality with a new refelxion of new Principles whose emerges of the basic ethic experiences. PMID- 18402230 TI - [The privacy respect and the professional secret in nursing]. AB - The situations that appear in health care demand keep awake aspects of the personal privacy, that in the normal life would be reserved to familiary relations. The interactions that settle down between the person who care and who is cared is an exceptional relationship of proximitiy. The healh professional, managers, computing pleople and administratives, collect original data of clinical history, these data are specially protected by the Protection Data law. Discretion has been considered an essential quality of the health professionals and were been collected in codes that regulate these professions. The arguments that defense the privacy value are bases in the person's value, his/her dignity and autonomy. PMID- 18402231 TI - [The informed consent in international clinical trials including developing countries]. AB - BACKGROUND: The informed consent procedure has been one of the most important controversies of ethical debates about clinical trials in developing countries. In this essay we present our recommendations about important aspects to consider in the informed consent procedure for clinical trials in developing countries. METHODS: We performed a full publications review identified by MEDLINE using these terms combinations: informed consent, developing countries, less developed countries and clinical trials. RESULTS: To protect volunteers in less developed countries should be valuated the importance of the community in the informed consent proceeding. The signing and dating of the informed consent form is not always the best procedure to document the informed consent. The informed consent form should be written by local translators. Alternative medias of communications could be needed for communicatios of the information to volunteers. CONCLUSIONS: Comparing with developed countries the informed consent proceeding in clinical trials in developing countries frequently require additional efforts. The developing of pragmatic researches is needed to implement informed consent proceedings assuring subjects voluntarily in each developing country. The main aspects to define in each clinical trial for each country are the influence of the community, the effective communication of the information, the documentation of the informed consent and local authority's control. PMID- 18402232 TI - [Ethics and efficacy in sexual health campaigns]. AB - The effectiveness of the diverse campaigns of sexual education carried out in Spain in the last 15 years scarcely has been analyzed. These campaigns have been directed fundamentally to adolescent people and its declared purpose has consisted of promoting the called "safe sex" being based only in the information on methods of barrier. Trying to clarify the efficacy of these campaigns, in the present work the epidemiological data contributed by the Department of Health and the National Institute of the Spanish Youth have been retrospectively investigated. As far as it can be measured, the evolution of the consequences of the sexual practices of the adolescents (abortions and unintended pregnancies) in the sanitary environment were also analyzed. Likewise, the data of the National Registry of Epidemiological Surveillance on the evolution of sexually transmitted diseases are collected. The results obtained of this analysis show that in teenages between 15 and 19 years, a progressive increase in the percentage of abortions regarding the total number of pregnancies has grown from 20% in 1990 to 44% in 2000 arriving at 46.6% in 2003. These data correspond with an progressive increase also related to the total number of abortions in Spain, reaching 13.7% in 2005. Likewise, the consumption of the postcoital pills for adolescents is analyzed. It has passed from 160.000 prescriptions in 2001 to nearly half a million units in 2005. This means the demand of this resource in the last 5 years has multiplied by three without achieving a stabilization in the number of new abortions per year. The evolution of the declared sexually transmitted diseases shows an increase of 79% in the infections by syphilis and a 45.8% in uncomplicated gonorrhoea. As conclusions, since an ethical perspective and since the perspective of sanitary efficiency, it can be affirmed that the validity of the campaigns of "safe sex" remains in doubt after analyzing the available data. The refusal to include in these campaigns the promotion of the abstinence in the first years of the adolescence, and the the refusal to promote the fidelity limiting the number of sexual partners, only seems to be justified for not sanitary, ideological motives, implying this a clear damage to the population at risk, saying nothing about the data of inefficiency, already available, of the campaigns previously carried out. PMID- 18402233 TI - [Respiratory symptoms in the general practice. An analysis of the Leipzig SESAM Study]. AB - The Saxon Study of General Medicine (SESAM) investigated the reasons why patients consulted the general practitioner, which diagnoses were established and how the patients were subsequently treated. In the majority of cases, the respiratory symptoms were simple infections of the upper and lower respiratory tract. Pneumonia must always be considered, while severe pulmonary diseases are of no significance in the differential diagnosis of respiratory complaints in the general practice. PMID- 18402234 TI - [Conjugated linoleic acids (CLA) and their relevance in the reduction of body fat. A critical review of the currently available data]. AB - Various experiments on animals have shown that conjugated linoleic acids (CLA) supposedly have numerous positive effects on health including reducing body fat. Although previous studies on humans did not lead to consistent results, the popularity of CLA as a weight loss product for overweight people and as a muscle building substance for athletes is increasing. Numerous internet providers or drugstores market CLA supplements or CLA-containing products. This article presents background information on the occurrence and biological effect of CLA and provides an up-to-date summary of studies on human subjects. Finally, whether supplementation with CLA for the reduction of body fat is useful will be evaluated. PMID- 18402235 TI - [Midlife crisis in men? Subjectively perceived physical and mental changes in men of advancing age]. AB - OBJECTIVE: What changes physically and mentally in men with increasing age? Does satisfaction with life only decrease? The personal subjective perception of the situation plays a decisive role in this assessment. One compares oneself to the past and to others, notices the illnesses of others and monitors oneself. METHODS: To study this problem, 85 men between the ages of 40 and 65 years were anonymously questioned by letter. The men filled out two questionnaires: one on change of life and virility symptoms (KLV) and a second on satisfaction with life (FLZ). The respondents were divided into three age groups: 40-47 (n = 26), 48-55 (n = 33) and 56-65 (n = 26) years old. RESULTS: The most frequent mental changes were related to concentration disorders (72.9%), tiredness (70.6%), increased irritability (65.9%) and a failing memory (63.5%). For physical changes, joint and limb pain (77.6%) were named the most frequently, followed by back pain (68.2%), the diminishing interest in sex (62.6%) and weight gain (55.0%). Whereas the rating of many criteria in the 40 to 47-year olds did not differ from that of the 56 to 65-year olds, the satisfaction with life in the 48 to 55-year olds in regards to leisure time, partner relationship, personal sexuality, satisfaction with oneself and social integration was lower. CONCLUSIONS: In summary, many areas of a man's life become destabilized at around age 50. Nevertheless, men are able to adapt to their changed physical and mental abilities and qualitatively reorient themselves. PMID- 18402236 TI - [The small difference in medicine. A survey on the acceptance of gender-related aspects in medicine]. AB - INTRODUCTION: Gender medicine investigates sex-related differences in morbidity and mortality. The consideration of relevant sex-specific factors should lead to an improvement in prevention, diagnosis and therapy. The goal of this survey was to obtain a picture of the current status and the future potential of gender medicine from the perspective of clinicians. METHODS: A questionnaire on gender medicine in education, diagnosis, therapy and research was sent to all professors as well as a random sample of assistant medical directors specializing in gynaecology, cardiology and neurology at German university hospitals. RESULTS: The response rate was 55% (N = 136). The present importance of gender medicine in Germany was rated by 83% of the respondents as low, and 62% said that gender medicine should be a required subject during medical studies. Through more consideration of gender-specific aspects, 72% expect a more exact diagnosis, 80% more specific treatment, 77% the avoidance of secondary complications, 64% a reduction of mortality and 73% the reduction of consequential costs. DISCUSSION: The current importance of gender medicine is assessed as rather low. The vast majority of the doctors expect clear improvements in patient medical care when the connection between gender and health are given more consideration. PMID- 18402237 TI - [The medical history as a historical source. As an example, the story of a war comrade of the "English patient", Count Laszlo Ede Almasy (1895-1951) is presented]. AB - The medical history is an important prerequisite for making a diagnosis and for establishing a trust- based doctor-patient relationship. Sometimes they can also serve as a historical source, as this example of an 89-year old patient shows. The patient worked with the Hungarian Count Laszlo Ede Almasy (1895-1951) during a German military secret service operation in North Africa during the Second World War. Graf Almasy became internationally famous through the novel "The English Patient" by Michael Ondaatje or rather, through the film of the same name. In the film however, the historical facts were very distorted as the medical history of our patient shows. PMID- 18402238 TI - [Does morphine have a life-shortening effect?]. AB - Morphine is a standard medication in pain therapy; opioids are grouped in step two and three of the WHO analgesic ladder. In palliative medicine, morphine is the preferred drug and is used for alleviating pain and reducing anxiety. When dosed correctly, a life-shortening effect can be definitely excluded today. Hence, the administration of morphine at the end of life cannot be called assisted suicide from a legal standpoint. PMID- 18402239 TI - [Can cardiovascular diseases be reduced over 80% by a "superpill"? Review on the epidemiological study from Wald and Law, 2003]. AB - As a consequence of our "western" diet and lifestyle, the risk for cardiovascular diseases is increased in the western world. It can be decreased by influencing the LDL-cholesterol values, the blood pressure and the platelet aggregation. It is, however, not proven that lowering of serum homocysteine levels reduces the risk forcardiovascular diseases. Wald and Law suggest in their epidemiological study a polypill, which has as its ingredients a statin, an anti-hypertonic drug, folic acid, and aspirine. Overall, the risk reduction by a polypill is overestimated, since a threshold is currently not existing, in all drugs side effects and interactions are possible, pharmaceuticals are only labelled for specific indications, and with missing efficacy there could be a negative risk benefit-relation. Indeed, a combined therapy with different drugs is in place in patients with higher risk and for secondary prevention, which is already risk adjusted. The use of effective drugs, however, in patients without evidence of a positive risk-benefit-relation and without medical control cannot be justified. Therefore, the application of the polypill for subjects beyond 55 years of age is an illusion; by contrast, treatment of patients with a high risk for atherosclerotic diseases is already reality. PMID- 18402240 TI - [Analgesic and muscle tonus normalizing effect of flupirtine retard in chronic back pain. Results of a standardized therapeutic evaluation applying objective methods for measuring pain pressure threshold, pain pressure tolerance and muscle tension]. AB - BACKGROUND: Chronic back pain is mainly caused by painful tension in the back muscles. Thus, analgesics with muscle tone decreasing effects that apparently normalize increased muscle tonus through specific modes of action without disturbing normal muscular movement are an important therapeutic option. Flupirtine retard (Katadolon S long) has provided such an option since 2006. OBJECTIVES: To impartially evaluate the muscle tonus normalizing effects of flupirtine retard by applying specific, objective test methods in patients with chronic musculoskeletal pain under routine practice conditions. METHODS: Prospective standardized evaluation of a treatment with flupirtine retard in 30 patients with continuous chronic, therapy refractory back pain. Measurement of general pain intensity, pain pressure threshold and pain pressure tolerance for trigger-point related pain and muscle tension in the affected back muscles before and during flupirtine retard treatment were performed in a standardized manner. RESULTS: In comparison to the reproducible, constant initial values, the two-week treatment with flupirtine retard led to a significant improvement in all measured muscle-specific indicators: pain pressure threshold (+48%), pain pressure tolerance (+27%) and depth of penetration in the muscle (+18%) (all values p < 0.001). These were also correlated with a clinically observable and statistically significant pain relief from an initial level of 7.0 +/- 1.3 to 3.0 +/- 1.4. CONCLUSION: Flupirtine retard was shown to be a useful, effective and very tolerable therapeutic option for patients with chronic back pain. The improvement of muscle disturbances which are responsible for the pain in addition to pain relief was shown firstly by objective measure methods. PMID- 18402241 TI - [Efficacy of a combination therapy with vitamins B6, B12 and folic acid for general feeling of ill-health. Results of a non-interventional post-marketing surveillance study]. AB - OBJECTIVE: In the present non-interventional postmarketing surveillance study, patients with symptoms of an inadequate supply of vitamins were tested for how a treatment with a combination vitamin injection consisting of vitamins B6, B12 and folic acid affects mood and fitness. The evaluation of the efficacy and tolerability as well as the documentation of adverse drug reactions were carried out by the physician. METHODS: The patient collective included 1430 patients (70.8% women, average age 67.1 years, average BMI 25.5 kg/m2). The average duration of treatment was 4.5 weeks with an average of 8.3 intramuscular injections. The principal method for determining the efficacy was the self assessment scale of well-being (Bf-S) according to Zerssen (sum score with a value range between 0 and 56 points). RESULTS: The sum score of the Bf-S decreased from 37.5 (+/-10.1) points at admission to 15.6 (+/- 9.4) points after four weeks of treatment. The subjective impression improved correspondingly in 96.3% of the patients. The improvement of the Bf-S was equally good in women and men. The number of injections correlated with the improvement in the sum score. The tolerability was mainly rated as very good or good. CONCLUSIONS: Eight vitamin infections over four weeks led to a clear improvement in the mood and vitality of patients with symptoms of intracellular vitamin B deficiency. PMID- 18402242 TI - [Investigation on the blood pressure-independent nephroprotective effect of nilvadipine in type 2 diabetics]. AB - BACKGROUND: The nephroprotective effect of ACE inhibitors and AT1 blockers has been demonstrated in manystudies. In particular, the effect produced by AT1 blockers has been described as being blood pressure-independent. The nephroprotective effect has been inconsistently observed in calcium antagonists; at the same time, it is always pointed out that calcium antagonists improve albumin excretion through a blood pressure-lowering mechanism. GOAL AND METHODS: Up to now there was not enough data available on the nephroprotective effect of nilvadipine. The goal of this study was to study the blood pressure-independent effect of the calcium antagonist nilvadipine on glomerular and renal tubular insufficiency during a 48-week treatment in hypertensive type 2 diabetics. Tubular or glomerular insufficiency was characterized on the basis of tubular and glomerular proteinuria and classified accordingto severity. RESULTS: Four of 24 patients showed an improvement in glomerular insufficiency after the 48-week treatment with a once-a-day dose of 16 mg nilvadipine. In 18 of the 24 patients, a progression in the disease was prevented. Renal tubular insufficiency was improved in 15 of 24 patients and progression in 20 of 24 patients was prevented. This nephroprotective effect on impaired tubular reabsorption, which has been ascribed to early damage in diabetic patients or as a possible prodromal stage of glomerular nephropathy in recent work, has not been observed in any other medicine. PMID- 18402243 TI - [Importance of a fixed combination of AT1-receptor blockade and hydrochlorothiazide for blood pressure lowering in cardiac risk patients. A postmarketing surveillance study with Candesartan/HCTZ]. AB - BACKGROUND: The treatment of arterial hypertension in Germany is, compared to international control rates, not adequate. In particular patients having additional cardiac risk factors like diabetes mellitus, dyslipidemia, increased waist circumference as well as concomitant diseases like myocardial infarction, heart- as well as kidney failure would benefit from an effective antihypertensive therapy. Aim of the present study was therefore to investigate in more detail blood pressure control in patients receiving a fixed combination of 16 mg Candesartan and 12.5 mg hydrochlorothiazide (HCTZ). METHODS: The present studywas performed as a non-interventional observational study. Included were patients with previously uncontrolled hypertension with at least one further risk factor. Primary variable was the blood pressure reduction over a time period of 8 weeks; secondary variables were the achievement of blood pressure targets and the tolerability of Candesartan/HCTZ. RESULTS: Between August 2006 and February 2007 3,787 patients in 893 physicians' offices in Germany were included. Patients were 62.2 +/- 11.3 years old, 48.1% were female, 97.5% had at least one additional risk factor, 29.8% a cardiovascular event. The risk to die from cardiovascular disease within the next 10 years was 7.4% according to the SCORE Score. By prescribing patients a fixed combination of 16 mg Candesartan/12.5 mg HCTZ a mean blood pressure reduction of -27.2/-13.4 mmHg was achieved (p < 0.001), pulse pressure was reduced by 13.8 mmHg (p <0.001) - both compared to previous therapy. A mean of 83.1% of patients achieved the guideline defined blood pressure targets (140/90 mmHg), 42.9% of patients even 130/80 mmHg. Serious adverse events were extremely rare (0.5%; hypertensive crisis, transitory ischemic attack). CONCLUSION: Prescribing patients with previously uncontrolled hypertension a combination of 16 mg Candesartan/12.5 mg HCTZ was tolerable and effective for blood pressure reduction in patients with arterial hypertension and additional risk factors like diabetes mellitus, dyslipidemia and the metabolic syndrome. PMID- 18402244 TI - Fetal cardiac surgery--a big challenge in the 21st century. PMID- 18402245 TI - Glucagon-like peptide 1 based therapy for type 2 diabetes. AB - BACKGROUND: Incidence of type 2 diabetes mellitus (T2DM) has increased in young people in recent years and new therapies are required for its effective treatment. Glucagon-like peptide 1 (GLP-1) is a potent blood glucose-lowering hormone produced in the L cells of the intestine. It may be potentially effective in the treatment of hyperglycemia in patients with T2DM. DATA SOURCES: PubMed database were searched with the terms "GLP-1", "incretins" and "diabetes". RESULTS: GLP-1 is a product of the glucagon gene, and its secretion is controlled by both neural and endocrine signals. GLP-1 lowers plasma glucose by stimulating insulin and suppressing secretion of glucagons, thus inhibiting gastric emptying and reducing appetite. GLP-1 exerts these actions by the engagement of structurally distinct G-protein-coupled receptors (GPCRs). In patients with T2DM, GLP-1 increases insulin secretion and normalizes both fasting and postprandial blood glucose when given as a continuous intravenous infusion. However, the native hormone is unsuitable as a drug because it is broken down rapidly by dipeptidyl peptidase IV (DPP-4) and cleared by the kidneys. Fortunately, many GLP 1 agonists or analogues and DPP-4 inhibitors have been found or developed, such as exendin-4, exenatide, liraglutide, CJC1131, vidaliptin and P32/98. Clinical trials have shown their therapeutic functions in T2DM with little adverse reaction. CONCLUSION: A GLP-1 based therapy will be safe and effective for the treatment of T2DM. PMID- 18402246 TI - Current concept about postoperative cholangitis in biliary atresia. AB - BACKGROUND: Postoperative cholangitis characterized by fever and acholic stool and positive blood culture is a common and serious complication following Kasai's operation for biliary atresia. The aim of this review was to describe the pathogenesis, clinical manifestations, medical treatment and outcome of postoperative cholangitis. DATA SOURCES: Articles on biliary atresia retrieved from Pubmed and MEDLINE in the recent 10 years were reviewed. RESULTS: The pathogenesis of postoperative cholangitis is still controversial. Recent methods for the diagnosis of postoperative cholangitis include urinary sulfated bile acids (USBA) and magnetic resonance cholangio-pancreaticography (MRCP). High-dose steroids and oral antibiotics have been used to reduce the incidence of postoperative cholangitis, and recurrent cholangitis leads to a lower survival rate. CONCLUSIONS: Cholangitis is one of the most important determinants of long term survival after the Kasai's procedure. The knowledge on postoperative cholangitis has been increasing in the past 10 years, showing a lower incidence of the disease and better therapeutic results. PMID- 18402247 TI - Vitamin A deficiency and child feeding in Beijing and Guizhou, China. AB - BACKGROUND: Vitamin A deficiency (VAD) is one of the three major micronutrient deficiencies in the world. In order to investigate the status of VAD and child feeding in China, we conducted the survey in Beijing city and Guizhou province. METHODS: We included a high socioeconomic area (Beijing) and a low socioeconomic area (Guizhou province) in China in our study. Participants included 1236 randomly selected children aged 0-71 months from stratified clusters (628 in Beijing and 608 in Guizhou), 409 from urban and 827 from rural areas. A food intake frequency questionnaire was used for dietary assessment. Fluorescence microanalysis was carried out to measure serum retinol concentrations. Serum retinol cut-off values of less than 20 microg/dl and 30 microg/dl were defined as sub-clinical VAD and suspected sub-clinical VAD, respectively. RESULTS: No xerophthalmia or night blindness was found. The mean concentration of serum retinol was 31.5 microg/dl in the high socioeconomic group, and 26.5 microg/dl in the low socioeconomic group. Rural infants had lower concentrations of serum retinol compared with the urban ones (26.9+/-8.1 microg/dl vs 31.8+/-7.3 microg/dl). The prevalence of sub-clinical VAD among all the children was 7.8%, and increased to 15.7% in children from the low socioeconomic group. In infants from the high socioeconomic area, the prevalence of suspected subclinical VAD was 38.0%, increasing to 59.5% in infants from the low socioeconomic area. The children from the low socioeconomic area had significantly lower fequency of intake of meat than the children in other groups. The prevalence of suspected sub clinical VAD was higher in the children with lower consumption frequency of vitamin A rich foods than the children with higher consumption frequency of vitamin A rich foods. CONCLUSIONS: VAD appears to be a moderate public health problem in certain areas of China. In areas with low socioeconomic status, VAD in childrean is more severe, and infants may be the group at the highest risk for VAD. Inadequate intake of vitamin A rich foods may result in VAD. A comprehensive long-term national strategy needs to be fostered in China for the treatment and prevention of the deficiency. PMID- 18402248 TI - Glucose metabolism disorder in obese children assessed by continuous glucose monitoring system. AB - BACKGROUND: Continuous glucose monitoring system (CGMS) can measure glucose levels at 5-minute intervals over a few days, and may be used to detect hypoglycemia, guide insulin therapy, and control glucose levels. This study was undertaken to assess the glucose metabolism disorder by CGMS in obese children. METHODS: Eighty-four obese children were studied. Interstitial fluid (ISF) glucose levels were measured by CGMS for 24 hours covering the time for oral glucose tolerance test (OGTT). Impaired glucose tolerance (IGT), impaired fasting glucose (IFG), type 2 diabetic mellitus (T2DM) and hypoglycemia were assessed by CGMS. RESULTS: Five children failed to complete CGMS test. The glucose levels in ISF measured by CGMS were highly correlated with those in capillary samples (r=0.775, P<0.001). However, the correlation between ISF and capillary glucose levels was lower during the first hour than that in the later time period (r=0.722 vs r=0.830), and the ISF glucose levels in 69.62% of children were higher than baseline levels in the initial 1-3 hours. In 79 obese children who finished the CGMS, 2 children had IFG, 2 had IGT, 3 had IFG + IGT, and 2 had T2DM. Nocturnal hypoglycemia was noted during the overnight fasting in 11 children (13.92%). CONCLUSIONS: Our data suggest that glucose metabolism disorder including hyperglycemia and hypoglycemia is very common in obese children. Further studies are required to improve the precision of the CGMS in children. PMID- 18402249 TI - Effect of phototherapy on blood endothelin and nitric oxide levels: clinical significance in preterm infants. AB - BACKGROUND: Phototherapy may have an adverse effect on the hemodynamics of preterm infants, and endothelin (ET) and nitric oxide (NO) are both the powerful vasoactive substances. This study was designed to observe the effect of phototherapy on blood levels of ET and NO in preterm infants. METHODS: Sixty-four preterm infants with hyperbilirubinemia requiring phototherapy were studied. Among them, 31 patients were born at 32-36 weeks' gestational age (GA), and 33 patients were < or =32 weeks GA. Control group included 26 full-term infants with hyperbilirubinemia requiring phototherapy. All patients were treated with continuous phototherapy for 24 hours. Blood samples were collected before and after phototherapy. The amount of ET in the blood samples was determined by radioimmunoassay, and NO levels were determined using nitrate reductase. Heart rate, respiratory rate, apnea, and mean arterial blood pressure (MABP) were monitored regularly (defined interval: hourly, 4 hours, etc) during phototherapy. RESULTS: Blood ET levels measured after 24 hours of phototherapy were higher than the pretreatment values, as were blood NO levels measured after 12 hours and 24 hours of phototherapy. Both increases were statistically significant (P<0.05) in the GA< or =32 weeks group. In the GA>32 weeks group, blood NO levels measured after 24 hours of phototherapy were higher than the pretreatment values; these changes were also statistically significant (P<0.05). In the GA<32 weeks group, heart rate increased and the MABP decreased during phototherapy. The changes after 24 hours of phototherapy compared to the pretreatment values were statistically significant. A few episodes of apnea occurred during phototherapy in the GA< or =32 weeks group. This was significantly higher than that in the other two groups. CONCLUSIONS: Under phototherapy, blood levels of ET and NO were significantly higher in preterm infants, especially in preterm infants of < or =32 weeks GA. PMID- 18402251 TI - Is pneumoperitoneum an absolute indication for surgery in necrotizing enterocolitis? AB - BACKGROUND: Necrotizing enterocolitis (NEC) is the most common gastrointestinal medical/surgical emergency in neonates. Non-operative support is needed in 70% of NEC cases, and surgical intervention in the rest 30%. Historically, pneumoperitoneum has been considered as an absolute indication for laparotomy. In the present study we emphasize that pneumoperitoneum is not an absolute indication for exploratory laparotomy in NEC cases. METHODS: We prospectively studied 58 patients with severe NEC having pneumoperitoneum on abdominal X-ray in the last 5 years. At the time of admission, the patients were given intravenous fluid, total parental nutrition, blood transfusion and broad spectrum antibiotics followed by abdominal tapping (paracentesis). All the patients with pneumoperitoneum were closely monitored for 48 hours if abdominal tapping was repeated. When the disease seemed to worsen clinically, radiologically and laboratorially, the patient was subjected to exploratory laparotomy. RESULTS: Of the 58 patients, 40 were treated conservatively whereas 18 underwent surgical intervention. The overall mortality in the present study was 12.1%, including 5% of the patients managed conservatively and 27% of the patients undergoing surgery. CONCLUSIONS: Pneumoperitoneum is not an absolute indication for surgery in cases of neonatal NEC. Most of the patients can be treated conservatively. PMID- 18402250 TI - Hemogram and bone marrow morphology in children with chronic aplastic anemia and myelodysplastic syndrome. AB - BACKGROUND: Aplastic anemia (AA) and myelodysplastic syndrome (MDS) are both acquired disorders in which bone marrow fails to produce or release sufficient blood cells. Anemia, infections and thrombocytopenia are common signs of such diseases. Clinically, it is difficult to distinguish chronic aplastic anemia (CAA) from MDS, especially from MDS without splenomegaly. As prognosis and treatment of AA and MDS are different, it is extremely important to make a differential diagnosis for the two diseases. METHODS: The medical records of 31 patients with CAA and 17 patients with MDS were retrospectively reviewed. Hemogram, bone marrow smear and biopsy for those patients were analyzed. RESULTS: The mean counts of monocytes and platelets in the peripheral blood of the CAA patients were significantly lower than those of the MDS patients. Bone marrow smear showed a reduction of cellularity in CAA patients. The mean counts of myeloblasts+promyelocytes, myeloblasts+proerythroblasts, and megakaryocytes in the bone marrow of CAA patients were markedly lower than those in MDS patients. But the mean lymphocyte count was reversed. Bone marrow cells showed morphological abnormalities in MDS. Hematopoietic tissue in the bone marrow biopsy decreased obviously in more than 96% of the patients with CAA. Adipose tissue in the bone marrow of CAA patients increased obviously. A reduction or deficiency (<2 cell/piece) of megakaryocytes was noted in 28 patients with CAA. Fibrous tissue in the bone marrow was detected in 5 patients with CAA. Bone marrow biopsy results showed hypercellular changes in 12 MDS patients. Ten patients showed aggregated erythroblasts which were in the same stage of development, and 15 patients had abnormal localization of immature precursors (ALIP). CONCLUSIONS: Blood cell counts can be decreased in addition to the reduction of cellularity in the bone marrow without dyshematopoiesis in CAA patients. Peripheral blood monocytes, fibrous tissue and cellularity in bone marrow are increased in MDS. Dyshematopoiesis and ALIP may appear characteristically in the children with MDS. Histology of bone marrow is important in the differential diagnosis of MDS and CAA. PMID- 18402252 TI - Functional constipation in children: investigation and management of anorectal motility. AB - BACKGROUND: Constipation is a common disease in children. Despite many causes, constipation is most often functional. This study was undertaken to investigate the anorectal motility in children with functional constipation as compared with healthy children and to determine the efficacy of management based on the results of anorectal manometry. METHODS: A multi-functional manometry was used to detect the anorectal manometry indexes of 8 patients with functional constipation (11.4+/-4.8 years) as well as those of 10 healthy children (10.5+/-3.5 years) from May 2004 to June 2005. The patients received a combined treatment regimen including probiotics (bifid triple viable bacterial tablet), prebiotics (lactulose) and regular defecation according to the results of anorectal manometry. The efficacy of these conservative measures was estimated during the course of treatment. RESULTS: No statistical difference was found in the indexes of effective length of the anal sphincter, anal tract maximal systolic pressure and the duration of more than 50% maximal systolic pressure between the two groups. But minimal sensitivity and maximal tolerated volume between the two groups were different significantly. Seven of the eight patients got better with the conservative treatment. CONCLUSIONS: Abnormalities exist in the anorectal motility of the children with functional constipation. Conservative treatment regimen based on the results of anorectal manometry is significantly effective. PMID- 18402253 TI - Enflurane requirement for blocking adrenergic responses to incision in infants and children. AB - BACKGROUND: Enflurane is one of the most commonly used inhaled anesthetics in China, but its requirement to block adrenergic responses after skin incision in pediatric patients is still unknown. This study was to determine the minimum alveolar anesthetic concentration (MAC) of potent inhaled anesthetics required to blunt the adrenergic response to skin incision of enflurane (MACBAR) in infants and children. METHODS: Twenty-eight patients, 10 infants (6-12 months) and 18 young children (1-6 years), were studied. The 18 children were randomly assigned into two groups, with or without fentanyl. Anesthesia was induced with 3 mg/kg propofol and 0.15 mg/kg vecuronium, and maintained with enflurane in 100% oxygen. Fentanyl (3 microg/kg) was given intravenously 5 minutes before incision for the patients of fentanyl group. The "up and down" method (with 0.3 MAC as a step size and 1 MAC as the start dose) was applied to determine MACBAR. The response was considered positive if the mean arterial pressure (MAP) or heart rate (HR) increased > or =15% after incision. The MACBAR was calculated as the mean of four independent cross-over responses in each group. RESULTS: MACBAR of enflurane in children of 1-6 years old was 3.2% (95% CI, 2.8%-3.6%) and was reduced to 2.2% (95% CI, 1.8%-2.5%) by 3 microg/kg fentanyl. In infants of 6-12 months old, the MACBAR of enflurane was 3.4% (95% CI, 3.0%-3.8%). CONCLUSIONS: MACBAR of enflurane in infants older than 6 months is similar to that in young children. The MACBAR of enflurane decreases with co-administration of fentanyl in the pediatric population. PMID- 18402254 TI - Considerations before repair of acquired rectourethral and urethrovaginal fistulas in children. AB - BACKGROUND: Acquired rectourethral or urethrovaginal fistula between the rectum or vaginal and lower urinary tract is an uncommon entity, which occurs as a consequence of pelvic disorder, including trauma, iatrogenic injury, inflammatory bowel disease, pelvic neoplasm or infection. But when is it appropriate to repair the fistula and what methods to be chosen? There has been no consensus on them. This study was undertaken to determine the timing of the procedure and the repair of rectourethral and urethrovaginal fistula. METHODS: From 1998 to 2006, we treated 19 children with rectourethral or urethrovaginal fistula, including rectourethral fistula in 15 boys and urethrovaginal fistula in 4 girls. The mean age of the patients was 6.2 years (range, 8 months to 11.5 years). The fistula occurred after pelvic fracture in 10 patients, and after iatrogenic injury in 9 including 4 after radical operation for Hirschsprung's disease and 5 due to anorectal malformation. Preoperatively, the general and local infections were controlled thoroughly, and complications such as urethral stricture and secondary megacolon were treated at first. At least 6 months after the last procedure, all patients underwent the 1-stage York-Mason procedure (via parasacrococcygeal incision) without colostomy and suprapubic cystostomy. Intraoperatively, the entire fistulous tract was excised completely. RESULTS: Infection and partial dehiscence of the wound occurred in 2 patients respectively. All fistulae were closed successfully without fecal incontinence or postoperative anal stricture. No patient suffered from urinary incontinence after fistula repair. The scars around the fistula were removed because they would shrink and lead to subsequent urethral occlusion or stricture. CONCLUSIONS: The timing of operation for acquired rectourethral or urethrovaginal fistula is appropriate at least 6 months after the last procedure. The 1-stage York-Mason procedure for the repair of the fistula is feasible and effective. PMID- 18402255 TI - Expression and clinical significance of stem cell marker CD133 in human neuroblastoma. AB - BACKGROUND: Recent evidences indicate that CD133, a kind of transmembrane protein, can be used as a marker to isolate stem cells from tumors originating from neural crest. This study was undertaken to explore the expression and clinical significance of stem cell marker CD133 in neuroblastoma (NB). METHODS: Immunohistochemical staining was used to detect the expression of CD133 in 32 patients with NB and 8 patients with ganglioneuroblastoma (GNB). The relationships were analyzed among CD133 expression, international neuroblastoma staging system (INSS) stages, pathological classification, and postoperative survival time of NB patients. RESULTS: The expression rates of CD133 in NB and GNB were 46.9% (15/32) and 37.5% (3/8) respectively, mainly in cytoplasm of neuroblastoma cells. The expression rates of stage 1-2, stage 3-4 and stage 4S were 30.7%, 57.9% and 37.5%, respectively. The differences in various stages were significant (P<0.05). The positive rate of CD133 in patients with unfavorable histology (52.4%) was significantly higher than that in patients with favorable histology (36.8%) (P=0.007). The survival time of CD133 negative patients was significantly longer than that of CD133 positive patients (P=0.026). CONCLUSIONS: CD133 which might be correlated with the development and progression of NB can serve as one of the important indicators for prognosis of NB. PMID- 18402256 TI - Esophageal achalasia of unknown etiology in infants. AB - BACKGROUND: Achalasia cardia is an uncommon disease in children particularly in infants. We present 8 cases of achalasia who were encountered over a 12-year period. In infantile achalasia, respiratory symptoms predominate and vomiting may easily be mistaken for gastroesophageal reflux (GER). Vomiting of uncurdled milk is characteristic of achalasia. METHODS: In this retrospective study, the data were obtained from records of the Department of Pediatric Surgery IMS, BHU Varanasi, India. The patients were diagnosed by clinical examination and barium study. Any other associated anomalies were noted in these patients. The patients underwent esophagocardiomyotomy with antireflux procedure via the abdominal route. RESULTS: In the present series, 7 patients survived. The follow-up study after operation showed remarkable relief of symptoms with satisfactory weight gain. No post-operative death occurred in the patients. Achalasia associated with alacrimia was noted in one infant who was lost to follow up. One infant was initially diagnosed as having gastric volvulus, but exploration revealed achalasia cardia. CONCLUSIONS: Esophageal achalasia is a rare disease in children and its origin is generally indeterminable. Achalasia cardia should be emphasized in the differential diagnosis of an infant presenting with signs and symptoms of esophageal obstruction. PMID- 18402257 TI - Dermoid cyst of the posterior fossa associated with congenital dermal sinus in a child. AB - BACKGROUND: Intracranial dermoid cysts are congenital benign neoplasms. Hydrocephalus and abscess as the principal manifestations of the posterior fossa dermoid cyst are rare. We present a case of obstructive hydrocephalus and abscess induced by an adjacent dermoid cyst with occipital dermal sinus. METHODS: A 2 year-old girl presented with headache and vomiting. Physical examination showed nothing abnormal except for a small subcutaneous nodule above the occipital protuberance with a small skin opening. She had no neurological deficits. Neuroradiological studies including CT and MRI showed a cyst located in the posterior fossa. The cyst in the posterior fossa with occipital dermal sinus was diagnosed. She was treated by radical excision of the occipital cyst through a suboccipital approach, and was followed up. RESULTS: Histopathologic examination suggested a dermoid cyst with an abscess. Bacterial investigation revealed Staphylococcus epidermidis, and appropriate systemic antibiotic therapy was given. The child recovered and a 2-year follow-up was uneventful. CONCLUSIONS: Posterior fossa dermoid cyst should be considered in all children with occipital skin lesions, especially dermal sinus. CT and MRI scan are helpful in the diagnosis of the lesion. Neurosurgical treatment of the lesion should be planned early to prevent infections such as abscess and meningitis. PMID- 18402259 TI - [Progressive low back pain in a young girl. Bending forward confirms the diagnosis]. PMID- 18402260 TI - [Every 3rd student has problems with alcohol. Intoxicated in the lecture hall (interview by Carola Goring)]. PMID- 18402258 TI - Diagnosis and treatment of blue rubber bleb nevus syndrome in children. AB - BACKGROUND: Blue rubber bleb nevus syndrome (BRBNS) is characterized by distinctive vascular malformations of skin and the gastrointestinal tract, often leading to chronic anemia and intestinal bleeding. It usually presents right after birth or during early infancy. Though the disease is inherent, its occurrence is sporadic. Thus it is usually not timely diagnosed. We analyzed the clinical characteristics and treatment of this disorder in order to improve the diagnosis and treatment. METHODS: Three patients with BRBNS treated at our hospital during 2002-2003 and 39 patients from the literature reported during 1965-2003 were reviewed in terms of the diagnosis and treatment. BRBNS may be diagnosed as cutaneous cavernous hemangioma associated with the same lesion of the gastrointestinal tract and other organs. RESULTS: Our 3 patients suffered from cutaneous angioma and gastrointestinal hemangioma. In 39 patients reported in the literature, cutaneous angioma was observed in all of them, and gastrointestinal hemangioma in 31. Additionally, the lesions were also found in other organs such as the brain (7 patients), joint (2), liver (2), eye (1), kidney (1) and spleen (1). Cutaneous angioma was located on the surface of the skin, including body (93%), limbs (86%), hip (36%) and face (26%). Gastrointestinal hemangioma was more common in the small intestine (100%) than in the colon (74%) and stomach (26%). When the joint was involved by hemangioma, pathologic fracture or overgrowth of bone needed traction and amputation (1 patient respectively). For significant gastrointestinal bleeding, endoscopic techniques (8 patients), surgical excision (5), or both (1) were performed. Recurrent bleeding was successfully treated by endoscopic laser combined with steroid or interferon in one patient. CONCLUSIONS: BRBNS in children presents atypical symptom and systemic complications. It should be dealt with seriously if gastrointestinal bleeding or orthopedic complication occurs. Treatment includes conservative, endoscopic and surgical options. Its recurrence with new angioma in the gastrointestinal tract needs laser-steroid therapy. PMID- 18402261 TI - [Where the tax auditor likes to investigate]. PMID- 18402262 TI - [Often the diagnosis is considered too late--family practitioners are also challenged. Many victims of sepsis could still be alive!]. PMID- 18402263 TI - [Losing no time in suspected sepsis. Every emergency bag should have a broad spectrum antibiotic! (interview by Dr. med. Horst Gross)]. PMID- 18402264 TI - [Etiology and therapy choices in dysphagia. Swallowing can be relearned]. PMID- 18402265 TI - [Significant dyspnea in a patient with COPD--ECG comparison pointed in the right direction]. PMID- 18402266 TI - [Rising prevalence, high cost. Hay fever remains a challenge]. PMID- 18402267 TI - [Allergen-specific immunotherapy in allergic rhinitis]. PMID- 18402268 TI - [ASA-intolerance (Samter's syndrome): an important differential diagnosis for chronic airway diseases]. PMID- 18402269 TI - [Norovirus gastroenteritis]. PMID- 18402270 TI - [Fournier gangene]. PMID- 18402271 TI - [Injury patterns in bombings]. PMID- 18402272 TI - [Emergency checklist: hypothermia]. PMID- 18402273 TI - 87% of hospitals aren't following all practices to stop avoidable infections. Report's findings are 'alarming'. PMID- 18402274 TI - Set a goal of zero central line and VAP infections. Determining what is really preventable. PMID- 18402275 TI - CDC: hospitals "need to do more" to control MRSA. AHRQ report disheartening. PMID- 18402276 TI - Discharge planning advisor. Program targets uninsured, underinsured patients. Goal: establishing 'home' at nearby center. PMID- 18402277 TI - Discharge planning advisor. CMS keeps emphasizing patients' right to choose. Discharge guidance refers to BBA. PMID- 18402278 TI - Core measures: are needed changes getting made? Concurrent data collection is best. PMID- 18402280 TI - Report: hospitals still struggling with many core measure requirements. PMID- 18402279 TI - Keep digging to uncover root causes. What do you do when your RCA hits dead end? PMID- 18402281 TI - Physicians may not report their impaired colleagues. PMID- 18402282 TI - Abbreviations: they're a definite threat to safety. PMID- 18402283 TI - Publicly reported data are misleading, says study. PMID- 18402284 TI - Your hand hygiene data may soon go 'public'. PMID- 18402285 TI - Implementing changes, overcoming resistance. PMID- 18402286 TI - Effects of olprinone on neuromuscular blockade caused by vecuronium. AB - We studied the effect of olprinone on neuromuscular blockade caused by vecuronium. Thirty women undergoing nitrous oxide-oxygen-isoflurane anesthesia were randomly divided into olprinone (n=15) or control group (n=15). In the olprinone group, the patients received an intravenous initial loading dose of olprinone at a rate of 2 microg/kg/minute for 5 minutes, followed by a continuous infusion of olprinone at 0.3 microg/kg/minute. In the control group, the patients received normal saline. Thirty minutes after the beginning of the infusion of olprinone or normal saline, vecuronium (0.1 mg/kg) was administered. The degree of neuromuscular blockade was monitored electromyographically at the adductor pollicis muscle. The time to the onset of neuromuscular blockade, and to the return of the first, second, third, or fourth response in train-of-four (TOF; T1, T2, T3, or T4, respectively), and the time course of recovery of T1/control did not differ significantly between the groups. After 50-70 minutes of vecuronium, the TOF ratio (T4/T1) in the olprinone group was significantly higher than in the control group. During this period, the mean TOF ratios in the control and olprinone groups were 0.15-0.39 and 0.40-0.57, respectively. In conclusion, olprinone accelerates the recovery of the TOF ratio, and the quickening effect of olprinone on the recovery of the TOF ratio may be apparent 50-70 minutes after vecuronium in anesthetized patients receiving vecuronium. PMID- 18402287 TI - Inhibitory oligodeoxynucleotide improves glomerulonephritis and prolongs survival in MRL-lpr/lpr mice. AB - Inhibitory oligodeoxynucleotides (ODNs), which are capable of blocking CpG induced inflammation, have been anticipated to be beneficial therapeutic agents for autoimmune diseases. In this study, we show that GpC ODN, which inverted the cytosine guanine sequence of CpG motif to guanine cytosine sequence, is an inhibitory ODN. The inhibitory effects of GpC ODN on CpG ODN-induced immune activation were confirmed by cytokine assay using splenocytes from lupus-prone MRL-lpr/lpr mice. In vivo, injecting MRL-lpr/lpr mice with GpC ODN did not reduce the deposition of IgG and C3 in the glomeruli, the serum level of IL-12, the serum level of rheumatoid factors and anti-ds DNA antibody, or alter the composition of IgG isotypes of anti-ds DNA antibody. However, the mice in the GpC group showed less proteinuria, significantly lower blood urea nitrogen levels (BUN) and significantly prolonged survival. Our results suggest that inhibitory ODNs, such as GpC ODN, have the potential to become a treatment for autoimmune diseases, like lupus nephritis. PMID- 18402288 TI - Can 3D-CT angiography (3D-CTA) replace conventional catheter angiography in ruptured aneurysm surgery? Our experience with 162 cases. AB - In this communication, we studied whether 3D-CT angiography (3D CTA) gives us enough information for a safe operation without those from conventional catheter angiography (CCA) in patients with ruptured aneurysms. Between December 1996 and September 2005, we treated 162 consecutive patients with ruptured aneurysms in the acute stage based on 3D-CTA findings. One hundred sixty-two ruptured aneurysms, including 64 associated unruptured aneurysms, were detected using 3D CTA. CCA was performed in nine (5.6%) of the 162 patients after 3D-CTA. They were four dissecting vertebral artery aneurysms, two basilar tip aneurysms, one basilar artery-superior cerebellar artery (BA-SCA), one previously clipped BA-SCA and one internal carotid-posterior communicating artery aneurysm. All ruptured aneurysms confirmed at surgery were treated successfully. The lack of information on CCA did not lead any neurological deficits or difficulties in the surgical procedure. 3D-CTA was of high diagnostic value compatible with CCA and yielded important information such as the configuration of the aneurysmal sac and neck, calcification in the aneurysmal wall, and the aneurysms' anatomic relation with adjacent vessels and bone structures. We suggest that 3D-CTA can replace CCA in the diagnosis of ruptured aneurysms and that most of ruptured aneurysms can be operated by using only 3D-CTA without CCA. PMID- 18402290 TI - Multiple and incidentally found cancers detected by 18F-FDG positron emission tomography/CT at one examination. AB - During the initial 8 months period of 18F-FDG PET/CT examination in our institution eleven cases of double cancers were detected. Eight cases were simultaneous second cancers and 3 cases are consecutive cancers. All cases are clinical ones and were referred from both outside hospitals and our own hospital. 18F-FDG PET/CT examination were utilized either to determine the extent of tumor or to stage the cancer or to detect recurrent tumors during the follow-up period. During the 8-months period 964 cases were studied. Therefore, the detection rates of simultaneous and consecutive cancers are 0.83% and 0.31% respectively. All together the detection rate of double cancer was 1.14%. To gain the general conception of double cancers the authors reviewed the autopsy registry of Japanese Society of Pathology during the four years from 2000 through 2003, and tabulated the combination of primary and second cancers. Frequently found combination of cancers were cancers of the thyroid, lung, stomach, liver, biliary tract, colon, rectum, and prostate. 18F-FDG PET/CT examination seems to be very useful in the management of cancer patients in terms of whole patient care. PMID- 18402289 TI - Breakthrough of immune self-tolerance to calreticulin induced by CpG oligodeoxynucleotides as adjuvant. AB - Reportedly, bacterial DNA containing unmethylated cytosine-guanosine dinucleotide motif-containing oligodeoxynucleotides (CpG-ODNs) can induce Th1-type adjuvant effects. We produced autoantibodies and induced hepatitis in mice using extracted proteins from human hepatocytes with CpG-ODNs as adjuvant. Western blot analysis was performed of sera from immunized mice and two patients with autoimmune hepatitis (AIH). When a common band was detected, N-terminal amino acid sequencing was performed to determine its site. For detection of antibodies against the identified protein (calreticulin), ELISA was performed of sera of 50 patients with AIH: 45 with primary biliary cirrhosis (PBC), 24 with chronic hepatitis C (CH), and 24 healthy controls. Mice were immunized with calreticulin protein with CpG-ODNs as adjuvant. Several reacted bands were detected in their sera; in addition, a common band to the sera of patients with AIH was detected at 60 kDa. Subsequent N-terminal amino acid sequencing revealed that the protein was human calreticulin. ELISA showed that, of patients with AIH, PBC, and CH, 30.0% (15/50), 17.8% (8/45), and 12.5% (3/24), respectively, were positive for anti calreticulin antibodies. Splenocytes from immunized mice produced IFN-gamma after they were pulsed with calreticulin protein. Histological analyses of liver specimens taken from mice immunized with calreticulin protein together with CpG ODNs showed spotty and focal necrosis. Immunofluorescence analysis showed increased expression of calreticulin in the liver treated with CpG-ODNs. These results suggest that a breakthrough of immune self tolerance to calreticulin is induced with CpG-ODNs as adjuvant and that calreticulin protein might be a target antigen in this model. PMID- 18402291 TI - Native troponin-T of the American cockroach (CR), Periplaneta americana, binds to IgE in sera of CR allergic Thais. AB - The American cockroach, Periplaneta americana, is the predominant cockroach (CR) species in Thailand and a major source of indoor allergens second only to the house dust mite. The incidence of CR allergy among allergic Thai patients is increasing but basic information on the allergenic components is scarce. In this study a recombinant troponin-T was produced by using cDNA prepared from RNA of the P. americana as a template and PCR primers designed from the P. americana troponin-T sequence deposited in the GenBank database. The recombinant protein (Mr approximately 50) did not bind to IgE in the sera of 18 skin prick test positive CR allergic patients. Rabbit polyclonal antiserum (PAb) against the recombinant troponin-T was produced and used in preparing an affinity column for the purification of native troponin-T from the crude P. americana extract (Mr approximately 47). IgE-immunoblotting revealed that the native protein bound to IgE in 3 of the 18 (16.7%) patients. Our results imply that native P. americana troponin-T, but not its recombinant counterpart, is a minor allergen among the CR allergic Thais. PMID- 18402292 TI - Bioinformatic studies on the group 2 allergens of Dermatophagoides farinae from China. AB - The present study aimed to characterize the group 2 allergens of the house dust mite Dermatophagoides farinae (Der f 2) from Hainan Island, a tropical region in Southeastern China. We cloned and sequenced cDNA coding for Der f 2 and found an additional region of 87 base pairs (bp) (from +77 to +163 bp) in our strain that was absent in the reference sequence (GenBank AB195580) used for primer design. However, the BLAST analysis identified the same sequence in strains reported from Reinbek, Germany, and Guangzhou, China. A phylogenetic tree was constructed using the Der f 2 nucleotide sequences from different regions or countries and showed that the Hainan sequence clustered with the strains from Reinbek and Guangzhou. Analysis of the translated amino acid sequence suggests that the encoded peptide is hydrophobic and extracellular with a cleavage site between the 17th and 18th amino acid residues and contains a strong trans-membrane helix from the 6th amino acid to the 24th amino acid, indicating a MD-2-related lipid recognition domain in this protein. Furthermore, the secondary structure of the pro-protein consists of 16.57% alpha helix, 32.57% extended strand and 50.86% random coil. In brief, we obtained a gene coding for Der f 2 and predicted the molecular characteristics of this protein using bioinformatics tools. Our analysis identified that this gene showed several significant differences to those reported previously. PMID- 18402293 TI - Adult-onset atopic dermatitis: a cross-sectional study of natural history and clinical manifestation. AB - The previously reported prevalence of adult-onset atopic dermatitis (AD) varied from 13% to 47%. There were a few reports of clinical features of adult-onset AD. The purpose of this article was to study the clinical features of Thai patients with adult-onset AD. We recruited prospective patients from the outpatient Department of Dermatology, Siriraj hospital, Mahidol University in Bangkok, Thailand, from June, 2006 to May, 2007. The diagnosis of AD was made according to the criteria of Hanifin and Rajka and the severity of AD in each patient was assessed using the Rajka and Langeland score. Fifty nine patients were enrolled. The majority of the patients developed their dermatitis during their third decade. Typical lichenified/exudative lesions were found in all cases. Non typical morphologic variants were found in 76.3%. The most common were nummular lesions. The main sites of involvement were the flexural area. The common sites of non-flexural involvement were the trunk, extensors and hands. The severity of AD was moderate in 64.4% of cases. A personal history of atopy was found in 84.7% of cases. Skin prick testing showed positive results (mostly to multiple allergens) in 25 of 29 patients (86.2%). Elevated serum total immunoglobulin E was detected in 6 of 10 patients (60%). It is concluded that adult-onset AD is not a rare but under-recognized eczematous condition. PMID- 18402294 TI - Orbital pseudotumor: clinical features and outcomes. AB - To describe the clinical characteristics of orbital pseudotumor, a retrospective analysis was performed on patients with orbital pseudotumor at Siriraj Hospital for ten years. Forty-nine patients (24 males and 25 females; 62 eyes) with a mean age of 43.75 years were included (a mean follow-up of 25 months). Thirty-six patients (73.5%) had unilateral disease. The clinical features were proptosis (79.6%), ocular motor deficit (61.2%), pain (51%), lid swelling or lid mass (44.9%), ptosis (24.5%), and chemosis (18.4%). The most common presenting sign was proptosis (49%). All were treated with corticosteroids with clinical improvement in 40 (81.6%) patients. Ten (83.3%) of 12 patients with visual loss improved with mean recovery time of 10.3 days. Ocular motility recovered in 24 (80%) patients, occurring an average of 17.8 days after initiation of therapy. It is concluded that the clinical features of orbital pseudotumor are varied. Most patients were improved with corticosteroids treatment. PMID- 18402295 TI - Seroprevalence of tetanus antibody in the Thai population: a national survey. AB - Tetanus is a disease with high mortality and the most important measure for effective prevention is vaccination. Tetanus immunization has been introduced to Thailand's national immunization program for 30 years. Yet, the coverage and seroprevalence of tetanus antibody in vast parts of the population has not been assessed. This study has been performed on 1,277 subjects aged between 6 months and 60 years or above from four geographically distinct provinces of Thailand. Tetanus antibody levels were measured using a commercially available ELISA kit. Most of the Thai population had immunity against tetanus. The level of antibodies to tetanus, as demonstrated by the geometric mean titer of antibody (GMT) (and 95% confidence interval) was 2.62 (2.34-2.91) IU/ml. The highest and lowest GMT was found in subjects aged between 5 and 9 years, and above 60 years of age with GMT (and 95% confidence intervals) of 3.64 (3.34-3.96) and 1.24 (0.67-2.29) IU/ml respectively. The minimum protective level of antitoxin (>0.01 IU/ml) was detected in 99.7 % of subjects. More than 90% of subjects displayed durable antibody protection levels (DAPL) (> or = 1.0 IU/ml), except for subjects above the age of 60 years (82%). According to this study, the majority of the population expresses tetanus antibody levels that can confer long term protection. Yet, considering the lowest GMT and the highest incidence of tetanus cases found in subjects aged above 60 years, re-immunization should be targeted at this age group especially if they had sustained any tetanus-prone injury. PMID- 18402296 TI - Components of pathogenic Leptospira spp. with potentials for diagnosis of human leptospirosis. AB - Existing serological methods for diagnosis of leptospirosis are still unsatisfactorily due mainly to their low accuracy. In this study, serum samples of 18 clinically diagnosed-, IgM dipstick positive-, MAT positive-leptospirosis patients (group 1) were analyzed by IgG Western blotting against SDS-PAGE separated-whole cell homogenates of pathogenic and non-pathogenic Leptospira spp. belonging to 20 serovars of 15 serogroups. The samples of group 1 were collected from the patients at days 3 to 10 after the fever onset (fist samples). Second and third samples could be obtained from 4 patients. Sera of the 22 patients with other febrile illnesses (group 2) and 22 healthy counterparts (group 3) were used as patient- and normal- controls, respectively. Irrespective of the serovar or serogroup of the pathogenic Leptospira spp. used as antigen in the Western blotting, all of the 18 sera of patients with leptospirosis (group 1) gave characteristic diffuse antigen-antibody reactive bands located at approximately 35-38 and 22-26 kDa; and thus 100% diagnostic sensitivity of the Western blot assay. Some serum samples of the leptospirosis patients also reacted to components located at 80-100, approximately 70, 60, 54, and 48 kDa. More bands or the early recognized bands with increased intensity were observed when tested the second and third samples. The characteristic bands were not seen when homogenates of L. biflexa, serogroup Semaranga, serovar Patoc (saprophytic) and L. biflexa, serogroup Andamana, serovar Andamana (non-pathogenic but can infect host) were used in the assay. Sera of groups 2 and 3 did not react to the components at the seven locations implying 100% diagnostic specificity of the IgG Western blot assay. While awaiting validation with more patients' samples, the IgG Western Blot analysis aiming at the detection of the characteristic antigen-antibody reactive bands described in this study has high potential for early, rapid, simple and accurate diagnosis of human leptospirosis. PMID- 18402297 TI - Cloning, protein expression and immunogenicity of HBs-murine IL-18 fusion DNA vaccine. AB - Hepatitis B is a global serious disease caused by hepatitis B virus (HBV). There is no known cure for hepatitis B. The best way to deal with the disease is by preventing with hepatitis B vaccine. However, the current protein-based vaccines made up of recombinant hepatitis B surface antigen (HBsAg) are ineffective in chronic HBV carriers and a significant number of the vaccinees do not mount the protective immune response. Novel DNA-based immunization may overcome the deficits of the protein-based immunization and may provide more effective prophylactic and therapeutic outcomes. In this study, we constructed a recombinant plasmid carrying gene encoding the HBV surface antigen (HBs) linked to DNA segment encoding full-length murine interleukin-18, i.e. pcDNA-HBs-IL-18. Immunogenicity of the DNA construct was carried out in BALB/c mice in comparison with mock, i.e. pcDNA3.1+ and vaccines comprised of pRc/CMV-HBs and pRc/CMV-HBs plus pcDNA-IL-18. All vaccinated mice revealed significant serum anti-HBs IgG response after two intramuscular injections of the vaccines at 28 day interval as compared to the level of mock. Co-administration of pRc/CMV-HBs and pcDNA-IL-18 elicited arbitrarily higher levels of anti-HBs IgG than the levels in mice immunized with pRc/CMV-HBs alone and mice that received pcDNA-HBs-IL-18 although not statistically different. Further experiments are needed to investigate the subisotypes of the IgG antibody, the kinetics of cytokine and the cell-mediated immune response. For this communication, the prototype HBs-IL-18 DNA vaccine was successfully constructed and the gene encoding murine IL-18 was successfully cloned. The latter can be co-injected with the antigen coding DNA or used as a fusion partner to the DNA for priming the immune response. The recombinant HBs and full-length IL-18 proteins have potential for other research purposes. They may be used also as standard proteins in the protein quantification assay. PMID- 18402298 TI - Nonsense mutations of the CYBB gene in two Thai families with X-linked chronic granulomatous disease. AB - X-linked chronic granulomatous disease (X-CGD) is an immunodeficiency disorder characterized by defective intracellular killing of microorganisms due to the neutrophils' inability to generate superoxide ions. Although it is always caused by mutations in the CYBB gene, clinical and molecular characteristics vary in different ethnic backgrounds. Two unrelated Thai boys presented with severe persistent pulmonary infections at the age of two months. Their abnormal dihydrorhodamine (DHR) flow cytometry assays supported the diagnosis of X-CGD. Mutation analysis was performed by polymerase chain reaction (PCR) amplification and sequencing of the entire coding regions of CYBB. Mutations identified were confirmed by restriction enzyme analyses. PCR-sequencing of the entire coding regions of CYBB identified nonsense mutations, 271C>T (R91X) in exon 4 and 456T>A (Y152X) in exon 5, in probands of each family. Both of the patients' mothers were found to be carriers. This observation supports that CYBB is the gene responsible for X-CGD across different populations and nonsense mutations are associated with severe phenotypes. PMID- 18402299 TI - A novel mutation of the CYBB gene resulting in severe form of X-linked chronic granulomatous disease. AB - We evaluated a boy who had multiple Salmonella septicemia, Aspergillus pneumonia and brain abscesses. His nitroblue tetrazolium (NBT) test was reportedly abnormal. The dihydrorhodamine (DHR) flow cytometry assay was compatible with typical X-linked chronic granulomatous disease (X-CGD). CYBB analysis revealed a novel complex mutation atggacg --> ttca in exon 12 (base pairs 1532-1538). As a result, 3 amino acids Tyr 511, Gly 512 and Arg 513 were deleted and replaced by 2 amino acids, Phe and Gln. The DHR and mutation analysis of his mother showed normal DHR pattern and no mutations in exon 12 of CYBB gene. In conclusion, any children with multiple Salmonella and Aspergillus infection should be suspected of CGD. NBT test, DHR assay and gene analysis are helpful toolsto confirm the diagnosis e v en i n the case of de novo mutation. PMID- 18402300 TI - Surgical approaches to malignant bowel obstruction. AB - The management of patients with malignant bowel obstruction (MBO) can be one of the most challenging aspects of advanced cancer care, and as a result, their symptoms are often palliated poorly, especially near the end of life. The term MBO encompasses a heterogeneous clinical syndrome,defined as obstructive symptoms due to the presence of intra-abdominal neoplastic disease. Radiological imaging, particularly with computed tomography, is critical in determining the cause of obstruction and possible therapeutic interventions. Options include laparotomy with or without a stoma, decompression with a stent, or aggressive medical therapy. Surgical decision-making involves the selection of the intervention most likely to relieve symptoms and improve quality of life for a particular patient at that particular point along his or her disease course. Although MBO is a relatively common dilemma encountered in clinical practice, there are no simple treatment guidelines or algorithms to follow. Instead, each patient must be assessed individually to devise a treatment plan that best balances the advantages and disadvantages of the intervention, considering the patient's prognosis, tumor biology, and-most importantly-his or her goals of care, as determined through an honest discourse between physician and patient. This review outlines a surgical framework for clinicians managing patients with MBO. PMID- 18402301 TI - Malignant bowel obstruction: tailoring treatment to individual patients. PMID- 18402302 TI - The value of a systematic approach to malignant bowel obstruction. PMID- 18402303 TI - Adjunctive nabilone in cancer pain and symptom management: a prospective observational study using propensity scoring. AB - A prospective observational study assessed the effectiveness of adjuvant nabilone (Cesamet) therapy in managing pain and symptoms experienced by advanced cancer patients. The primary outcomes were the differences between treated and untreated patients at 30 days' follow-up, in Edmonton Symptom Assessment System (ESAS) pain scores, and in total morphine-sulfate-equivalent (MSE) use after adjusting for baseline discrepancies using the propensity-score method. Secondary outcomes included other ESAS parameters and frequency of other drug use. Data from 112 patients (47 treated, 65 untreated) met criteria for analyses.The propensity adjusted pain scores and total MSE use in nabilone-treated patients were significantly lower than were those found in untreated patients (both P < 0.0001). Other ESAS parameters that improved significantly in patients receiving nabilone were nausea (P < 0.0001), anxiety (P = 0.0284) and overall distress (total ESAS score; P = 0.0208). The nabilone group showed borderline improvement in appetite (P = 0.0516). When compared with those not taking nabilone, patients using this cannabinoid had a lower rate of starting nonsteroidal anti inflammatory agents, tricyclic antidepressants, gabapentin, dexamethasone, metoclopramide, and ondansetron and a greater tendency to discontinue these drugs. PMID- 18402304 TI - A coordinated, multidisciplinary approach to caring for the patient with head and neck cancer. PMID- 18402305 TI - Benign hypertrophy of the prostate. PMID- 18402306 TI - Treatment of benign prostatic hyperplasia at Mater Hospital, Nairobi. AB - OBJECTIVE: To evaluate the use of medical therapy in the management of patients with mild, moderate and severe symptoms of benign prostatic hyperplasia. DESIGN: Prospective study. SETTING: Private urology clinic in Mater Hospital, Nairobi between 1995 and 2005. PATIENTS: Six hundred and eighty patients with benign prostatic hypertrophy aged 50-80 years. RESULTS: Two hundred and eighty (43.8%) patients benefited from the drug treatment alone and were released from the clinic follow up after five years. Two hundred and seventy (42.2%) cases could not complete the five years follow up on drugs alone due to symptoms recurrence or severity or unavailability of drugs. They were removed from the drugs alone treatment and put on other interventions. Ninety (15.9%) cases were operated on based on their decision and insistence or severity of their symptoms or no drugs response. CONCLUSIONS: This observational study confirms that medical treatment is beneficial in the management of benign prostatic hyperplasia (BPH) and is cheap and gives better first line option of treatment choice to many patients. It also gives hopes to the surgical risk patients. The quality of life (QOL) of patients and their family disturbance is also improved. PMID- 18402307 TI - Comparison of four-day and ten-day post-mastectomy passive drainage in Accra, Ghana. AB - OBJECTIVES: To determine the optimum period for passive post-mastectomy drainage in Accra, Ghana, comparing early (day 4) to late (day 10) drain removal. DESIGN: Randomised prospective clinical study. SETTING: Surgical Department, Korle Bu Teaching Hospital, Accra, Ghana SUBJECTS: Patients with breast cancer scheduled for total mastectomy and level 11 axillary clearance. INTERVENTIONS: Patients were randomised to one of the two groups: early (day 4) and late (day 10) passive drainage. MAIN OUTCOME MEASURES: Duration of hospital stay, seroma formation and postoperative wound infection. RESULTS: Forty five patients had 4-day drainage and 42 had 10-day drainage. Late removal of drains (Day 10) was associated with a significantly higher drainage (1123ml) than those with early (4-day) drain removal (571 ml); p=0.0019. Late removal, however, had fewer seromas (28.6% vs. 46.7%; p=0.2), smaller aspirate volumes (435ml vs. 563ml) and fewer number of aspirations (2.1 vs. 3.2). Early removal had a shorter hospital stay of 6.2 (+/ 1.52) days compared to 11.07 (+/- 0.76) days for late removal, and a lower wound infection rate (2.2% vs. 9.5%). There was a low incidence of seromas when drainage was <30 ml/day at the time of drain removal. CONCLUSIONS: Post mastectomy passive drains may be removed when drainage is <30 ml in 24 hours. When there is a persistent large volume of axillary drainage, patients should be counselled about the relative risks of early and late drain removal. For patients who do not find seroma aspiration unacceptable, early (day 4) removal appears preferable to late (day 10) removal of drains. PMID- 18402308 TI - Exfoliative erythroderma at Kenyatta National Hospital, Nairobi. AB - BACKGROUND: Exfoliative erythroderma (EE), (Synonyms: Exfoliative dermatitis, Red man syndrome) is a clinical syndrome characterised by generalised erythema and scale. It is an important cause of functional skin failure and associated high morbidity and variable mortality rates. OBJECTIVES: To study demographics, aetiology, complications and clinical outcomes of exfoliative erythroderma (EE) on patients attending Kenyatta National Hospital (KNH). DESIGN: Cross- sectional descriptive study. SETTING: Kenyatta National Hospital, Dermatology Unit. SUBJECTS: All available medical records on inpatients seen by qualified dermatologists at KNH with generalised erythema and scale from 1996 to 2006. MAIN OUTCOME MEASURES: Discharge or death. RESULTS: Incidence exfoliative erythroderma was documented in 146 out of all 123 admissions (13%) into the dermatology unit from 1996-2006. Demographic mean age was 47 years, M: F ratio was 3:2, 67% had no income and 53% and 30% were residents of Nairobi and adjacent districts respectively. Sixty three percent were due to skin diseases, 23% due to systemic diseases of which 20% were due to HIV/AIDS and 14% due to adverse cutaneous drug reactions. Ninety percent of patients were treated and discharged and 10% died; 50% of whom had dermatoses and 29% due to HIV associated antituberculous drugs. CONCLUSIONS: Exfoliative erythroderma is an important cause of morbidity, admission and mortality in patients attending KNH. Dermatoses and HIV / AIDS were the most frequent causes. The mortality rate was relatively low and attributable to controllable diseases. PMID- 18402309 TI - Self-reported adherence to single dose nevirapine in the prevention of mother to child transmission of HIV at Kitale District Hospital. AB - OBJECTIVES: To evaluate the uptake and adherence to single dose nevirapine among HIV positive mothers. DESIGN: Descriptive cross-sectional study. SETTING: The maternal and child health and family planning (MCH-FP) clinics in Kitale district hospital, Western Kenya. SUBJECTS: HIV positive postnatal women attending MCH-FP clinic who had gone through the PMTCT programme. RESULTS: A total of 146 respondents were recruited for this study. Most (90%) of them reported swallowing their nevirapine tablets, however only 55 swallowed their tablets within 4-12 hours before delivery. The most important factor affecting nevirapine adherence was place or delivery (p<0.05). Most (71%) of mothers who did not swallow their nevirapine delivered at home. Women attending ANC for two times or less young women under 20 years of age and single women were also less likely to swallow their nevirapine (p < 0.05). Most (91%) of the babies received their nevirapine syrup with 98% of them getting it within 72 hours of delivery. Eighty eight percent of babies who did not take their nevirapine were delivered at home. Babies whose mothers did not take their nevirapine were also more likely to miss it. CONCLUSIONS: Self reported adherence to take home nevirapine is high. However mothers who deliver in a health facility were more likely to access nevirapine both for themselves and their babies than those delivering at home. PMID- 18402310 TI - Assessment of outpatient therapeutic programme for severe acute malnutrition in three regions of Ethiopia. AB - OBJECTIVE: To document the experiences and lessons for rolling out of the OTP service at the wider scale with the aim of assessing the strengths and weaknesses of the project and suggest recommendations for future programming. DESIGN: Qualitative methods of data collection including focus group discussion, observation and in-depth interview of key informants were employed to get relevant data. Review of health facility, reports and programme documents were done to capture further- information. SETTING: Out Patient Programme (OTP) pilot programme implemented by CONCERN/ VALID in three administrative regions of Ethiopia namely: South Nations and Nationalities Peoples Regions (SPNNR), Addis Ababa and Oromia regions. A total of thirteen health centres which had started OTP service from the three regions were included in the study. SUBJECTS: Thirty six key informants and 30 focus group discussants were involved in the study conducted from 16th to 25th November 2006. RESULTS: Out Patient Programme (OTP) has enhanced community's understanding of malnutrition as a health problem through an excellent entry point it created for behaviour change communication (BCC) on optimal infant and young child feeding (IYCF). It has also enhanced utilisation of the existing equipments of the respective health services to promote nutrition and increased mental satisfaction of the providers who observed rapid recovery of malnourished children taking the plumpy nut. It also resulted in increased awareness of the community about malnutrition and its treatment, which resulted in increased need-based demand for the OTP and self-referral of children to health facilities. Shift in the thinking of the providers on the fact that malnutrition can be treated without admitting the child and reduction in the burden of malnutrition and associated mortality are other positive findings of the study. CONCLUSION: While it was observed that the programme was very effective in treating case of severe acute malnutrition and is highly acceptable by planners, health care providers and beneficiaries, there were different operational issues that needed to be strengthened. The irregularity and incompleteness of supply availability, high attrition of trained human power, inadequate supportive supervision especially from local ministry of health, inadequate community mobilisations are some of the shortcomings identified. Based of these findings recommendations were forwarded. PMID- 18402311 TI - Aetiology, diagnosis and management of haemospermia: a review. AB - OBJECTIVE: To provide an overview of the aetiology, investigations and the various treatment methods currently available in the management of haemospermia. DATA SOURCE: Review of literature was effected through medline and index medicus search of major published indexed journals and books. DATA SELECTION: Published data on haemospermia, hemospermia, haematospermia and semen over the last five decades (1967-2007) were utilised. DATA EXTRACTION: Abstracts of selected articles were read and analysed to determine their possible contribution and relevance to this article. DATA SYNTHESIS: All relevant articles were reviewed in full and contribution extracted for this review as necessary. CONCLUSION: Haemospermia (haematospermia) is a relatively frequent, distressing and frightening symptom in sexually active men. It is usually a benign self-limiting condition resolving within several weeks except for the few with underlying aetiology including prostate malignancy and idiopathic. Patients presenting with haemospermia should have a detailed medical history, physical examillation including blood pressure measurement, genital and digital rectal examination. Persistent and recurrent haemospermia is best investigated by TRUS, CT, MRI, urethrocystoscopy, and biopsy and histological confirmation of malignancies. Specific treatment depends on the underlying pathological cause but often involves only minimal investigations and simple reassurance. PMID- 18402312 TI - Cheilitis glandularis progressing to squamous cell carcinoma in an hiv-infected patient: case report. AB - Cheilitis glandularis (CG), is a rare inflammatory minor salivary gland disease affecting the lower lip. The hallmarks of which include progressive enlargement and eversion of the lower labial mucosa resulting in the obliteration of the mucosal-vermillion interface. A case is presented of a 47-year-old HIV-infected woman who initially manifested clinical features of CG with a typical histopathology picture of a non-specific sialadenitis while a second biopsy performed six months later revealed well differentiated squamous cell carcinoma (SCC). She is symptom free one year following excision and radiotherapy treatment. PMID- 18402313 TI - Conservative management of horizontal root fracture of a maxillary permanent incisor: case report. AB - A case is presented of an 11-year-old boy who was diagnosed with a horizontal radicular fracture located in the middle third of a maxillary right central incisor that was immediately managed conservatively. Subsequent to the injury, the patient complained of inability to bite, a swollen upper lip and pain on palpation in the periapical region of the tooth. Clinically the tooth appeared extruded. Radiographic examination revealed a root fracture. The extruded tooth was repositioned and immobilised with a wire-composite splint, under a suitable prophylactic antibiotic and anti- inflammatory analgesic coverage for one week. Two years after trauma, the treated tooth was firm and healthy with good aesthetics and normal function. PMID- 18402314 TI - Infective endocarditis. PMID- 18402315 TI - [Imperforate hymen; importance of inspection of the external genitalia]. AB - Three women aged 17, 12 and 13 years presented with intermittent abdominal pain. Secondary sexual characteristics, i.e. mammae and pubic hair, were developed, but menarche had not occurred. All were found to have imperforate hymen. After incision, the symptoms disappeared. Imperforate hymen causes clinical symptoms from the time of menarche. Pubertal girls presenting with abdominal symptoms must be examined, including observation of secondary sexual characteristics and inspection of the external genitalia. This examination is often omitted because the patient and the physician are embarrassed. However, it is essential and should be carried out in a considerate manner. Excision of the hymen must be performed under general anaesthesia using a technique that will prevent secondary stenosis. PMID- 18402316 TI - [The value of follow-up in surgical oncology]. AB - Follow-up after surgical interventions for cancer is essential for the early detection of recurrent (2nd) primary and metastatic disease; patient coaching, including psychosocial coaching; and treatment quality control. As the incidence of cancer increases, cost-benefit analyses become increasingly important. The most effective period of follow-up is 5 years. An intensive follow-up schedule has been shown to increase life expectancy in colorectal cancer patients through early detection and timely intervention for recurrent disease. In melanoma patients, self-detection plays an important role. Nurse practitioners are increasingly involved in follow-up to reduce costs and specialist time. A differential follow-up schedule that takes into account the initial tumour stage can further enhance the cost-benefit ratio of follow-up. PMID- 18402317 TI - [The practice guideline 'Asthma in children' (second revision) from the Dutch College of General Practitioners; a response from the perspective of general practice]. AB - The second revised guideline 'Asthma in children' from the Dutch College of General Practitioners adequately describes the present state of knowledge regarding the diagnosis and treatment of asthma in children. Updates include the recommendation to investigate allergies in children aged less than 6 years, because this can contribute to a probable diagnosis of asthma. This specific recommendation, which has yet to appear in international guidelines, is based on an excellent clinical study performed in general practices in the Netherlands. It is important that the findings of this study are confirmed by at least one additional study. The revised guideline correctly asserts that fast-acting bronchodilators and inhaled corticosteroids form the cornerstone of treatment for children with asthma, and that cromoglycate no longer has a role in this setting. However, the question remains whether the currently modest role assigned to leukotriene antagonists is justified based on the most recent literature. PMID- 18402318 TI - [The practice guideline 'Asthma in children' (second revision) from the Dutch College of General Practitioners; a response from the perspective of paediatric pulmonology]. AB - The second revised guideline 'Asthma in children' from the Dutch College of General Practitioners provides a well-documented and useful overview. Diagnostic recommendations depend heavily on the symptom 'wheezing', which may be difficult to detect in young children. A uniform therapeutic approach for the various asthma phenotypes is recommended, including a therapeutic trial with inhaled corticosteroids for persistent symptoms. Pitfalls include the fact that inhalation of corticosteroids is sometimes difficult in young children, and that the use of corticosteroids in children with transient virus-induced wheeze is not evidence-based. The guideline no longer recommends the use of cromoglycate. Children who are refractory to inhaled steroids should be referred to a paediatrician before trying long-acting beta2-sympathomimetics or leukotriene antagonists. The current challenge is to effectively incorporate this new guideline into a coherent system of care for the asthmatic child in cooperation with paediatricians, asthma nurses and others involved in the diagnosis and treatment of childhood asthma. PMID- 18402319 TI - [Practical questions related to self-measurement of blood pressure]. AB - The results of self-measurements of blood pressure predict the risk of developing cardiovascular disease better than those of blood pressure measurements taken at the GP surgery or hospital. In spite of the increasing availability of devices for home measurement, exactly how, by whom, with what and when, blood pressure should be measured at home remains unclear. Self-measurement is to be recommended as a supplement to conventional blood pressure measurement, as, in this way, the white-coat effect and masked hypertension can be recognized. Self-measurement is only useful if it is carried using a validated, automatic, sphygmomanometer and measured in the correct way. It is essential that the patient be clearly instructed on how to do this. A limit for home measurement of 135/85 mmHg should be adhered to. When blood pressure measurements taken at home lead to a different conclusion than those taken at hospital or GP surgery (and if there is no white coat or masked hypertension), it is recommended that the procedure be repeated. If after this, there is still a discrepancy between the results of these two methods of blood pressure measurement, ambulatory 24-hour blood pressure measurement will perhaps provide the definitive answer to the 'real' level of the patient's blood pressure. PMID- 18402320 TI - [Summary of the practice guideline 'Asthma in children' (second revision) from the Dutch College of General Practitioners]. AB - A number of important changes have been made in the second revision of the guideline 'Asthma in children' from the Dutch College of General Practitioners. In children under the age of 6 years, the symptoms stuffiness and recurrent cough are no longer considered part of the symptomatic diagnosis of asthma. Wheezing has become the key symptom of asthma. In children aged 6 years or more, spirometry is the optimal method for both diagnosis and monitoring. This method is preferred over peak flow measurement. Inhalation allergies should be investigated in children under the age of 6 years because the presence of an inhalation allergy may influence the management approach. Starting asthma medication in children under the age of 6 years should always be considered a therapeutic trial, and its effect should always be evaluated. The prescription of allergen-resistant mattresses and bed coverings is only effective when it is one component of a set of allergen reduction measures. At this time, the Dutch Health Council recommends influenza vaccination in children with asthma. PMID- 18402321 TI - [Diagnostic image (364). A boy with a painful inguinal mass]. AB - A 9-year-old boy presented with right-sided abdominal pain and an inguinal mass due to torsion of an acquired ectopic testicle. PMID- 18402322 TI - [Detection of the first recurrence in patients with melanoma: three quarters by the patient, one quarter during outpatient follow-up]. AB - OBJECTIVE: To evaluate the benefit of frequent outpatient follow-up after the initial diagnosis and treatment of melanoma. DESIGN: Retrospective study. METHOD: Patients from the Groningen University Medical Centre with a first recurrence ofa previously treated primary melanoma (American Joint Committee on Cancer stage I III) were interviewed to determine how many of them had detected the recurrence themselves. Patient data and recurrence characteristics were compared with data from a previous Australian study in order to evaluate the differences between the Dutch and the Australian population. RESULTS: 70 patients with a first recurrence of melanoma were studied. Of the 70 first recurrences, 53 (76%) had been detected by the patient; 10% of the patients had a sign or symptom, but did not detect the recurrence themselves; 11% of the first recurrences were detected as a consequence of self-examination. The proportion of detection by the patient versus detection by a doctor was roughly equal in the Dutch and Australian populations. CONCLUSION: Three quarters of the first recurrences of a treated melanoma had been detected by the patients themselves, the largest number by accident and only 11% by self-examination. It is not likely that the continuation of frequent follow-up visits will contribute to the detection of recurrences. These findings are no different from the results in the Australian population. PMID- 18402323 TI - [Low complication rate with cataract operations carried out by registrars in ophthalmology]. AB - OBJECTIVE: To determine the complication rate of cataract operations and learning curve of registrars in ophthalmology. DESIGN: Retrospective, descriptive investigation. METHOD: 982 consecutive cataract operations performed by registrars in ophthalmology at University Medical Centre Utrecht and the Meander Medical Centre, Amersfoort, the Netherlands, were evaluated for complications. Of patients in whom a complication occurred, the nature of the complication, any possible follow-up operation, and best corrected visual acuity before and following operation were assessed, as well as the stage of training the registrar was at. RESULTS: The most important complications occurred in the following frequencies: endophthalmitis 0%, posterior capsule rupture with vitreous loss 1.32%, dropped nucleus 0.2%. Four patients required a second operation. There were no statistically significant differences in complication rate between the three phases of the training. In all patients with complications the best corrected visual acuity was more than 0.5. CONCLUSION: The complication rate was smaller than described in the literature and comparable with rates known for ophthalmologists. Training in cataract surgery does not lead to irresponsible risks for the cataract patient. The combination of training in both a university hospital and a general hospital has several advantages for a registrar in ophthalmology. PMID- 18402324 TI - [Two neonates with congenital aniridia: the necessity of genetic investigation]. AB - Two female neonates were diagnosed post partum with bilateral aniridia. The first patient had the familial form, caused by a point mutation in the paired box 6 (PAX6) gene. The second patient had a sporadic aniridia caused by a de novo microdeletion involving both the PAX6 gene as well as the Wilms tumour suppressor I (WT1) gene. This made screening for the presence of a Wilms tumour necessary. The second patient died several months after birth, due to respiratory insufficiency. Aniridia is a rare developmental disorder of the eye, with absence of most of the iris tissue, caused by an abnormality in the PAX6 gene on chromosome 11p13. Familial aniridia is usually due to a point mutation of the PAX6 gene, which causes solely ocular abnormalities. Sporadic aniridia is caused by a de novo deletion or microdeletion of chromosome 11p13, which affects not only the PAX6 gene but also the adjacent WT1 gene. In these patients, the Wilms tumour, aniridia, genitourinary anomalies, and mental retardation (WAGR) syndrome can be present, and screening for a Wilms tumour is indicated. Unless previous investigation of a family member has demonstrated the WT1 gene to be normal, chromosome studies should always be performed in patients with aniridia. PMID- 18402325 TI - [Viewpoint of the Dutch Melanoma Working Group regarding sentinel lymph node biopsy: prognostic but not therapeutic implications]. AB - The sentinel lymph node procedure provides prognostic information for patients with cutaneous melanoma, but its therapeutic implications were unclear until now. The international multicentre selective lymphadenectomy trial has shown that sentinel lymph node biopsy confers no survival benefit for patients with cutaneous melanoma. The presence of melanoma cells in a sentinel lymph node, however, has prognostic value. Therefore, the Dutch Melanoma Working Group recommends discussing this procedure with all patients with melanoma of Breslow thickness 1.2-3.5 mm. There is no evidence to suggest that the sentinel lymph node procedure is beneficial when the Breslow thickness is less than 1.2 mm or greater than 3.5 mm. PMID- 18402326 TI - [Skid marks]. PMID- 18402327 TI - [Of old people.....]. PMID- 18402328 TI - [Alternative flow chart for laboratory procedures for anemia]. PMID- 18402329 TI - [Alternative flow chart for laboratory procedures for anemia]. PMID- 18402330 TI - [Alternative flow chart for laboratory procedures for anemia]. PMID- 18402331 TI - [Calcium-sensing receptor. From the bench to the bedside and to the public health]. PMID- 18402332 TI - Endoscopic treatment of high-risk bleeding ulcers: success, rebleeding and mortality. AB - INTRODUCTION AND AIMS: Endoscopic treatment of peptic ulcers with high-risk stigmata has been probed. The rates of recurrent bleeding, need for emergent surgery and death are related to Forrest Classification, Blatchford's modified risk score and the kind of endoscopic treatment used (monotherapy vs. dual). The aims of the present study were to report the success of endoscopic therapy in the reduction of the rate of initial success, recurrent bleeding, the need for surgery, and the mortality rate for patients with bleeding peptic ulcer and high risk stigmata. PATIENTS AND METHODS: From a retrospective view, patients seen from September 2004 to March 2007 who had peptic ulcers Forrest Ia, Ib, IIa and/or IIb were included. RESULTS: Fifty-six patients were included (mean [SD] age 57.3 +/-16.6 years). The success rate was 91%, whilst the rest of the patients required immediate surgery. Recurrent bleeding was presented in 14 (27%) patients and eight (14.2%) required emergency surgery. The mortality rate was 3.6%. No factors were associated with the risk of failure to initial treatment, recurrent bleeding or need for surgery. The use of monotherapy by endoscopy was associated with the mortality. The variable "performed by a fellow alone" was not associated with any kind of outcome. CONCLUSION: Complication rate is similar to previous reports of general hospitals, but is higher than those of referral centers. Endoscopic monotherapy is associated with a major mortality risk. PMID- 18402333 TI - A single-institution, 20-year prospective experience with an affordable Fc receptor blockade method to treat patients with chronic, refractory autoimmune thrombocytopenic purpura. AB - In a 20-year period in a single institution, 34 patients with chronic, refractory autoimmune thrombocytopenic purpura were prospectively treated with ex vivo anti D opsonized autologous red blood cells. All patients had received previous treatment with steroids and/or immunosuppressive agents, and 11 had been splenectomized. Twenty one patients had an increase in the platelet count; in five cases, the increase was more than 50 x 10(9)/L platelets and in 16 the increase was more than 100 x 10(9)/L platelets. Early responses were observed in 20 patients and late responses in seven, whereas seven patients (20%) did not respond at all. Nine of the 20 individuals who achieved an ER had a subsequent drop in the platelet count; however, only three had a drop below 50 x 10(9)/L. When last censored, of the 34 patients, 24 (70%) had a platelet count above 50 x 10(9)/L. The 84-month thrombocytopenia-free (over 50 x 10(9)/L platelets) status of the whole group is 70%, whereas the 84-month complete remission (over 100 x 10(9)/L platelets) status of the whole group is 50%. It is concluded that the use of ex vivo anti-D opsonized red blood cells may represent another, substantially cheaper treatment of patients with chronic, refractory, autoimmune thrombocytopenic purpura. PMID- 18402334 TI - [Quality of medication prescribing for elderly patients with non-malignant pain syndrome attending family medicine clinics]. AB - OBJECTIVE: To assess the quality of medication prescribing in ambulatory elderly patients attending to family medicine clinics due to non-malignant pain syndrome. PATIENTS AND METHODS: By doing a secondary data analysis we evaluated the quality of medication prescribing in 495 subjects aged 60 or more years, which were seen due to non-malignant pain syndrome in family medicine clinics belonging to Instituto Mexicano del Seguro Social. The analyzed information included general patient characteristics, medical histories, and medication (complete data). The quality of medication prescribing was assessed by using the Beers criteria and the MAI. All data were analyzed by using descriptive statistics. RESULTS: The total number of medicines prescribed was 3017, with an average number of prescribed drugs of 6 (+/- 2.5). About 35% of patients had prescriptions with at least one inappropriate medication according to the Beers criteria. The mean MAI score was 3.5 (+/- 4.3) for drug and 20.9 (+/- 10.7) per patient. According to MAI criteria, the most frequent errors were: failure to provide practical directions, prescription of drug combinations with potential drug-drug interactions, and inappropriate indications. CONCLUSION: Inappropriate prescription is a common problem in elderly patients with nonmalignant pain syndrome attending to primary care facilities. PMID- 18402335 TI - [Prevalence of Salmonella sp. in foods in Tamaulipas, Mexico, during 2005]. AB - OBJECTIVE: To determine the prevalence of Salmonella sp. in foods obtained from Tamaulipas sanitary jurisdictions during 2005. MATERIALS AND METHODS: Several kinds of foods were taken from the State of Tamaulipas from January to December 2005, which were analyzed in the Public Health State Laboratory of Tamaulipas. If the food fulfilled with the acceptance criteria, it was analyzed looking for Salmonella sp. in agreement with NOM-114-SSA1-1994. Positive foods were sent to the National Institute of Epidemiological Reference and Diagnostic (INDRE) to determine Salmonella species and serovariety. Data were collected in a database and analyzed using chi2 and Fisher's exact test in STATA 8.2 software. RESULTS: 24 of 1300 analyzed foods were positives to Salmonella sp. (1.9%, CI 95% 1.2 2.7). By sanitary jurisdiction, Victoria showed the highest prevalence of positive results (3.9%). When the analysis was carried out by "north jurisdictions" versus "south jurisdictions", south jurisdictions showed a high prevalence of Salmonella in foods (0.9% vs. 2.5%, p < 0.05). Sausage was the most contaminated food (20.0%). A higher percentage of positive results was noticed in terrestrial foods than in sea-derived foods (2.3% vs. 0.3%; p < 0.05). In January (5.5%) and May (3.7%) the highest prevalence of positive results was found; without significant difference by year seasons (p > 0.05). The most common bacterial species was Salmonella enterica, serovariety enteritidis, which was found in 58.3% (n=14) of positive foods. CONCLUSIONS: The jurisdictions with the highest prevalence of Salmonella in foods were identified. Terrestrial foods showed more Salmonella contamination. The results show that thanks to epidemiologic surveillance it is possible to identify potential sources of salmonelosis outbreaks. A bigger research effort it is necessary to extent the epidemiologic surveillance activities and their results, by one side toward the kind of foods initially identified as "high risk foods", and by other side, to other regions of Mexico in order to know the distribution of this pathogen and establish adequate preventive measures. PMID- 18402336 TI - Duplication 2p and monosomy 8p in mosaicism: clinical, molecular cytogenetic and molecular markers of a unique case. AB - We report on a female patient, with a de novo mosaicism for a structural rearrangement producing trisomy 2p21-->pter and monosomy 8p21-pter. GTG bands and fluorescence in situ hybridization (FISH) in lymphocytes identified: mos 46,XX,der(8)(8qter-->8p21::p21::2p21-->pter),9qh +[52]/ 46,XX,9qh+[82]. Fibroblasts showed the same cell lines in 15 and 12 cells respectively. DNA profiling with fourteen autosomal STR markers, did not reveal a chimerism status in our patient. She did not present the classical phenotype described for trisomy 2p and for monosomy 8p probably due to approximately 60% of the patient's cells being normal. The abnormality probably arose in a very early stage of development during the first post-fertilization divisions with a non-sister chromatid exchange event between chromosomes 2 and 8 producing three cellular clones: a normal clone, one with trisomy 2p and monosomy 8p and a third with monosomy 2p and trisomy 8p. Only the first two cell lines were found in both lymphocytes and fibroblasts of hypopigmented skin; the third may have been lost or limited to other tissues. PMID- 18402337 TI - [Critical reading of clinical research reports among resident doctors at IMSS of five specialties. Exploratory national study]. AB - INTRODUCTION: Critical reading holds an important role in methodical criticism of clinical aptitude, which contributes to its perfection, which would be indispensable in the study and consideration in the continuing formation of the medical resident. OBJECTIVE: To investigate the degree of development of critical reading of information of clinical investigation, in medical residents of five specialties in the country. MATERIAL AND METHODS: A cross-sectional study took place, in 3060 medical residents of five residencies (internal medicine n=474, pediatric n=358, family medicine n=1651, ER/doctor surgical n=269 and anaesthesiology n=308). In order to measure the degree of development of critical reading, an instrument with eight clinical research article summaries was elaborated, selected according to the proposed classification of types of studies by McMaster University, with subjects that could be of interest for the studied residents; constituted by 150 items that explore three indicators (to interpret, to judge and to propose); validated by five experts with educational experience, clinical-epidemiologist and in publication of works; a pilot study was made and the internal trustworthiness with the test of Kuder-Richardson was considered (KR 20), obtaining a 0.90 coefficient. The capture of data and the qualification of the instrument were made with the blinded technique. The analysis was made with parametric statistic. RESULTS: The residencies that received greater points were internal medicine and pediatrics (X 50.23 +/- 15.02 and X 48.84 +/- 15.76, respectively, of a theoretical qualification maxima of 150). Also, it was not observed in the points reached in the critical reading by the resident doctors, an ascending tendency per year of residency (of R1 to R4). CONCLUSIONS: In critical reading, residents of internal medicine as well as pediatrics obtained the highest scores in contrast with family medicine, ER, surgical and anesthesiology which obtained lower scores. Also, it seems to be that until this moment, the development of this aptitude does not keep correspondence as it is passed per year of residency. PMID- 18402338 TI - [AMPK as a cellular energy sensor and its function in the organism]. AB - The adenine monophosphate (AMP) activated protein kinase (AMPK), is a heterotrimeric complex that is activated by an increase in the AMP/ATP ratio, and is considered to be a cellular energy sensor that contributes to regulate energy balance and caloric intake. AMPK is activated by LKB1 hinase and it can phophorylate several enzymes involved in anabolism to prevent further ATP consumption, and induces some catabolic enzymes to increase ATP generation. Furthermore, AMPK regulates the expression of genes involved in lipogenesis and mitochondrial biogenesis, among others. AMPK is distributed in most organs including, liver, skeletal muscle, heart and hypothalamus; and even in adipose cells. In addition, AMPK is activated in the hypothalamus stimulating appetite due to energy depletion. AMPK also participates in glycolysis regulation, glucose uptake, lipid oxidation, fatty acid synthesis, cholesterol synthesis and gluconeogenesis, and it has been considered as a possible target enzyme in the treatment of some diseases such as obesity, type 2 diabetes and hepatic steatosis. This review provides a general overview of AMPK structure, its activators and its function in the organism. PMID- 18402339 TI - [Pathophysiological role of aldosterone in acute renal failure]. PMID- 18402340 TI - [Toll-like receptor in lung response to pathogens]. AB - Innate immunity plays a central role in antimicrobial defense. Advances in the understanding of pathogen recognition systems of innate cells have yielded the identification of Toll like receptors (TLR) as key elements of the lung defense mechanisms which is heavily exposed to a variety of stimuli. TLR recognition of several microbial compounds induces proinflammatory cytokines production whose contribution to the host may be either protective or detrimental. Human immune response diversity may explain the differences observed between patients facing bacterial, viral and fungal lung infections. New strategies designs that modify innate immune response may be useful to limit detrimental consequences of inflammatory processes in the lung. PMID- 18402341 TI - [Letter of Dr. Lister to Dr. Arista]. PMID- 18402342 TI - Analysis of haptoglobin phenotype polymorphism in a Mennonite population of northern Mexico. PMID- 18402343 TI - Spirituality and nursing. PMID- 18402344 TI - A historical overview of spirituality in nursing. AB - The following article is an attempt to encapsulate an historical overview of spirituality in nursing. Despite a plethora of information relative to spirituality in nursing, the decision was to do an eclectic overview that was not reflective of any one spiritual/religious group. The authors at times found this goal difficult when writing about the Pre-Christian and Christian eras. Most of the major religions have their own perspective on the concept of spirituality, and historical personalities recorded maybe reflective of that particular religion. Another factor that impacted on the writing of this article was the concept that spirituality was not always linked specifically to religion. Spirituality was an experiential component of the wonders of nature and life, a domain that the authors took into account but did not expound upon. Finally the authors realized that spirituality has almost always been an aspect of African American life and certainly of those in nursing. To this end the authors realize that there is a need for more research in this area. PMID- 18402345 TI - Spirituality in the natural sciences and nursing: an interdisciplinary perspective. AB - The Big Bang theory, a widely accepted theory of the origin of the universe, states that the universe was created between ten to twenty billion years ago from a cosmic explosion. Charles Darwin, a 19th century English naturalist, convinced the scientific community through his work that life evolved by natural selection over three and a half million years ago rather than through the influence of a Supreme Being or creator. Although there has been scientific data to support the claims of natural selection, there still remain many unanswered questions suggesting that other mechanisms contributed to the evolution of life. These unresolved findings greatly influenced mysticism and the development of the theological argument, which suggest the existence of a supreme being (God), who is believe to be an omnipotent healer, comforter, provider of salvation, and the center of mysticism spirituality. There has been consistent use of spiritual practices to address health concerns by individuals for thousands of years. There is increasing data that supports the implementation of spirituality in nursing for client care to enhance health outcome and patient wellbeing. Incorporating spiritual care into practice is an integral dimension of holistic care that is the crux of nursing practice in the 21st century. Holistic care of clients requires that nurses use the nursing process to implement spiritual care in practice. PMID- 18402346 TI - Integrative review of spirituality in African American breast cancer survivors. AB - This paper reports findings of an integrative review of the literature on spirituality in AA breast cancer survivors, isolates key spiritual themes, and recommends future research. Inclusion criteria are 1994 to 2004 research studies that included AA breast cancer survivors 18 years old and older. Content analysis was used to isolate spiritual themes and spiritual domains/dimensions. Seven studies resulted that used qualitative, quantitative, and mixed methods. Themes identified were spirituality provided the strength to cope; the need to care for others and receive care; beliefs that God is the healer and in control; God assists in decision-making; and closeness to God. Spiritual domains were beliefs, functions, and social support. Future research should explore the domains/dimensions and meanings of spirituality experienced by diverse groups ofAA breast cancer survivors. Culturally appropriate, evidence-based nursing care should include spiritually based interventions that acknowledge the significance of God. PMID- 18402347 TI - Spiritual well-being, religious coping, and the quality of life of African American breast cancer treatment: a pilot study. AB - There is a dearth of knowledge about the quality of life of African American women during the breast cancer treatment phase. This pilot study explored spiritual wellbeing, religious coping, and the quality of life of African American women during the breast cancer treatment phase. The sample included a total of 11 African American women from the mid-Atlantic and southeastern United States. The Roy Adaptation Model (Roy & Andrews, 1999) served as a guide for this study. This pilot study used a descriptive cross-sectional design. Data were analyzed utilizing descriptive statistics and the Spearman rho correlational analysis. African American women used more positive religious coping than negative religious coping. Significant relationships were found between spiritual well-being and the QOL domains of physical, emotional, and functional well-being. These findings suggest that nurses should incorporate spiritual and religious support in the care of African American women during the breast cancer treatment phase. PMID- 18402348 TI - Assessing spirituality in mentally ill African Americans. AB - The case scenario illustrates the advantage of using spirituality as a tool for recovery when working with mentally ill African American clients. Often spiritual and clinical perspectives are seen as contradictory. But for African Americans, these perspectives can be mutually reinforcing. Spirituality can serve as a resource of strength. It can provide emotional consolation, inspiration, guidance, and security. It can foster personal responsibility, identity, respect for ethical codes and community building. Mental Health professionals who use spirituality as a tool for recovery can expect to have better client outcomes when working with African Americans than those who do not. PMID- 18402349 TI - The influence of spirituality on health care-seeking behaviors among African Americans. AB - This study examined spirituality in the cultural context of African Americans and how it relates to their health care-seeking behaviors. Focus group methodology was utilized for data collection. Three focus (3) groups were conducted in the Hampton Roads area. The sample consisted of African American men (n=12, 35%) and women (n=22, 65%). Between 11 and 12 participants were in each group for a total of 34 participants, ranging in age between 10 and 80 years. A semi-structured interview guide facilitated the focus groups. This guide was designed to facilitate the subjects' overall perceptions of spirituality, health and health care-seeking behaviors. Focus group data were transcribed and analyzed by selecting phrases exemplifying common themes. RESULT: The findings indicated that spirituality had an influence on health and health-care seeking behaviors in African Americans. It was concluded that spirituality should be a focus for health care professionals in providing holistic care for African Americans. PMID- 18402350 TI - The influence of spirituality and religiosity on breast cancer screening delay in African American women: application of the Theory of Reasoned Action and Planned Behavior (TRA/TPB). AB - African American women (AAW) are 25% more likely to present with late stage breast cancer and 20% more likely to die from their disease than Caucasian women. Researchers report that a treatment delay of 3 months is a significant factor in breast cancer mortality. Socioeconomic factors, lack of access and knowledge, spiritual and religious beliefs, fear and fatalism are reported as contributing factors to screening delays. Studies have primarily applied the Health Belief Model (HBM) and modified versions like the Champion HBM to preventive health practices. Neither have significant inclusion of spirituality or religiosity. The TRA/TPB focus on beliefs, intent and attitude as individual determinants of the likelihood of performing a specific behavior; but have not had wide utility in studies related to screening delays among AAW. This paper explores the utility of applying the TRA/TPB as the theoretical framework for determining cultural relevance of spirituality and religiosity to screening delays among AAW. PMID- 18402351 TI - Measuring maternal responsiveness. AB - The purpose of this study was to review current research studies examining maternal responsiveness to locate self-report instruments, and to identify variables in these self-report instruments used to describe maternal responsiveness. The electronic databases, Medline, PsychLit, SocLit, and Cinahl were searched with the following key words: maternal responsiveness, maternal sensitivity, maternal attachment, maternal response, and responsiveness. Articles were obtained, assessed, and agreed upon for their usefulness to the study by the researcher and research assistant. Criteria included: articles must include key word, maternal responsiveness, in the title or a self-report instrument measuring maternal responsiveness. Fifty-two eligible studies were identified from the literature, read by two reviewers and both reviewers confirmed its applicability. All included studies were summarized in tabular form. References with the key words in the title were extracted from the literature to examine the measurement strategies. Specifically, the researchers were looking for self-report measures of maternal responsiveness. The trials identified by this review addressed limited aspects of communication and focused solely on antenatal care. Current self-report instruments do not contain the variables of maternal awareness, maternal recognition, and maternal reflection. Future research is needed in this area of instrumentation and maternal responsiveness. PMID- 18402352 TI - Teaching the psychology of women: an interdisciplinary approach. AB - This paper explores the development of a college freshman level course entitled, "Psychology of Women" at the University of the Incarnate Word in San Antonio, Texas. Interdisciplinary focus groups were held that included female professors from various disciplines, i.e., business, education, English literature, biology, nursing, and religious studies. The critical question posed to the group was: In this era of globalization and diversity, what issues and topics should be covered in a class that explores the psychology of women across the lifespan? The importance of studying womanhood within the context of nationality, religion, race, ethnicity, socioeconomic status, age, and physical ability was highlighted. PMID- 18402353 TI - Outcomes from Walk the Talk: a nursing intervention for Black women. AB - OBJECTIVE: Explore changes in walking behavior, blood pressure, and weight over the course of a pilot intervention designed to increase physical activity. DESIGN: A pre/post single group design was used to examine outcomes of a 12-month group intervention with 6-month follow-up. SAMPLE: Twenty-one sedentary hypertensive African American women residing in Mid-Missouri with a mean age of 50.3 years. INTERVENTION: A 3-hour monthly meeting was paired with an at-home walking component. RESULTS: Mean steps per day taken by participants were 3857 at baseline, 4060 at the end of the group intervention, and 5282 at the end of the follow-up period. Mean systolic blood pressure was 142 mmHg at baseline, 123 mmHg at intervention end, and 130 mmHg at 18 months. Mean diastolic blood pressure at baseline was 87 mmHg, 80 mmHg at 12 months, and 78 mmHg at the end of the follow up period. Mean weight decreased from 206 lbs at baseline to 188 lbs at the end of the intervention. However, mean weight had increased to 213 lbs by the end of the 6-month follow-up. CONCLUSIONS: Outcomes from this pilot study are encouraging with respect to the potential of group level interventions as a strategy for promoting cardiovascular health among hypertensive African American women. It is particularly noteworthy that women maintained decreases in systolic blood pressure and further decreased diastolic blood pressure despite regaining weight during the follow-up period. Replication of this study with a larger sample size and control group will significantly enhance our ability to evaluate whether the unusual pattern of change in walking behavior noted in this pilot was a fluke or indicative of a slower pattern of change in this population. PMID- 18402354 TI - College females as mothers: balancing the roles of student and motherhood. AB - As more female college students are involved in sexual relationships their risk of conception increases. However, when pregnancy occurs it is only the woman who bears the burden and risk of the pregnancy and in most cases child care. Female college students who become pregnant are then faced with, not only the risk of the child birth but also the responsibility of their education and childcare. This study describes how the women in situations like this cope with their experiences. In their own words the participants will suggest how they handle their responsibility of school and parenting. Five themes are associated with this study: (a) mother's unconditional love; (b) relationship with the child's father; (c) responsibility for education; (d) family and friends involvement; and (e) the learned lessons. These themes are used as a guide to get an understanding of what the females go through when faced with these responsibilities. PMID- 18402355 TI - [Clinical and neurophysiological aspects of depression and apathy]. PMID- 18402356 TI - [RTMS and depression: what is offered in the literature?]. PMID- 18402357 TI - [Psychopharmacotherapy in psychiatric comorbidities among patients with chronic pain]. PMID- 18402358 TI - [Dimensional structure of psychoses: relations with diagnostic categories and contribution into the study of endo-phenotypes]. PMID- 18402360 TI - [Drugs acting on D3 receptors in neuropsychiatric disorders: facts and prospects. Implication for schizophrenia and its treatment in the regulation of BDNF dependence on D3 receptor expression]. PMID- 18402359 TI - [Epigenetic model in the pharmacologic modulation of vulnerability to schizophrenia]. PMID- 18402361 TI - [Compulsive buying: psychological and biological treatment]. PMID- 18402362 TI - [Hedonistic homeostasis in Parkinson disease]. PMID- 18402363 TI - [Ouality indicators for the evaluation of colorectal cancer screening programmes]. AB - Colorectal cancer screening programmes, which are supported by national and international recommendations, are gradually developing andspreading throughout Italy. The programmes strive to guarantee maximum equality, improve quality of life, and inform the target population about benefits and possible risks. These goals can only be achieved with adequate planning and training for the staff involved. Moreover, it is essential to regularly verify the quality of each step of the process, with a system that can constantly monitor the parameters used for the evaluation of quality and efficacy. The present document, created by the Italian Group for Colorectal Screening (GISCoR), proposes a unified method for collecting and reporting screening data using commonly agreed terminology, definitions and classifications, thus making it possible to compare data with standards and data from other programmes, and to plan suitable interventions. The indicators considered refer to the entire screening process, including organisational, logistic and performance indicators. The indicators are provided in the form of concise and easy-to-use data sheets. Every data sheet contains briefsections: definition of the indicator, goal, the data necessary to build it, the formula required to build it, possible problems of interpretation, acceptable and desirable standards. PMID- 18402364 TI - PDA survey of quality risk management practices in the pharmaceutical, devices, & biotechnology industries. AB - In July 2006 the Parenteral Drug Association's Risk Management Task Force for Aseptic Processes, conducted an electronic survey of PDA members to determine current industry practices regarding implementation of Quality Risk Management in their organizations. This electronic survey was open and publicly available via the PDA website and targeted professionals in our industry who are involved in initiating, implementing, or reviewing risk management programs or decisions in their organizations. One hundred twenty-nine members participated and their demographics are presented in the sidebar "Correspondents Profile". Among the major findings are: *The "Aseptic Processing/Filling" operation is the functional area identified as having the greatest need for risk assessment and quality risk management. *The most widely used methodology in industry to identify risk is Failure Mode and Effects Analysis (FMEA). This tool was most widely applied in assessing change control and for adverse event, complaint, or failure investigations. *Despite the fact that personnel training was identified as the strategy most used for controlling/minimizing risk, the largest contributors to sterility failure in operations are still "Personnel". *Most companies still rely on "Manufacturing Controls" to mitigate risk and deemed the utilization of Process Analytical Technology (PAT) least important in this aspect. *A majority of correspondents verified that they did not periodically assess their risk management programs. *A majority of the correspondents desired to see case studies or examples of risk analysis implementation (as applicable to aseptic processing) in future PDA technical reports on risk management. PMID- 18402365 TI - Development and evaluation of push-pull based osmotic delivery system for pramipexole. AB - An oral push-pull system that can deliver pramipexole for extended period of time has been developed and characterized. A bilayer osmotic drug delivery system was developed using a basic design consisting of an oral controlled porosity osmotic pump. Unlike other osmotic systems, which require a preformed orifice for drug release, controlled porosity membranes contain water-soluble pore-formers in the coating membrane. When such systems come in contact with water, the additives dissolve resulting in an in-situ formation of a microporous membrane. The push layer swells releasing the drug at a controlled rate. In advanced Parkinson's disease the usual dose of pramipexole is 1.5 mg three to four times a day. Hence, an attempt was made to develop a once-a-day controlled release system. This may offer significant patient benefits by providing enhanced efficacy and reduced side effects and may also reduce the number of daily doses compared to conventional therapies. This developed push-pull system was compared with other types of osmotic delivery systems, such as an asymmetric membrane coating and a dense coat with mechanical drilling. An optimized system was selected to study the effect of the concentration of a pore-forming agent such as PEG 400 and dibutyl phthalate, the pH of dissolution media, the effect of agitation and osmotic agents on drug release. The osmotic pressure generated was determined using a 3D3 freezing point osmometer. The drug release was found to follow zero order kinetics. Drug release increased with an increase in osmotic pressure. The developed push-pull osmotic system showed the desired once-a-day release kinetics. PMID- 18402366 TI - Preparation, characterization, and pharmacokinetic evaluation of puerarin submicron emulsion. AB - A novel formulation of puerarin was studied. Puerarin submicron emulsion was prepared by complex phase inversion-high-pressure homogenization technology. Characterization, distribution of drug in emulsion, short-term stability, and pharmacokinetics of emulsion were evaluated. The mean diameter and zeta potential of puerarin emulsion were 188.14 nm and -29.45 mv, respectively. The distribution range of puerarin emulsion was very narrow. The concentration of puerarin in the interfacial surface, oil droplet, water, and liposome-micelles were 7.821, 1.079, 0.637 and 0.423 mg/mL, respectively. Puerarin submicron emulsion was stable for a period of 3 months. The area under the whole blood concentration-time curve of rabbits after intravenous administration of puerarin emulsion was 1.718-fold higher than that of rabbits of intravenous administration of puerarin (P < 0.05). And compared with the puerarin group, the elimination rate of puerarin emulsion group was significantly decreased (P < 0.05), and the biological half-life and the mean retention time of puerarin emulsion were markedly increased (P < 0.05). PMID- 18402367 TI - Nanoemulsion system for the transdermal delivery of a poorly soluble cardiovascular drug. AB - The purpose of the present study is to develop and evaluate the potential of nanoemulsions for increasing the solubility and the in vitro transdermal delivery of carvedilol. Pseudoternary phase diagrams were developed and various nanoemulsion formulations were prepared using oleic acid and isopropyl myristate (IPM) (1:1) as the oil, Tween 80 as surfactant, and Transcutol P as cosurfactant. The prepared nanoemulsions were subjected to physical stability tests. Transdermal permeation of carvedilol through rat abdominal skin was determined with Keshary-Chien diffusion cell. Significant increase (P < 0.05) in the steady state flux (Jss) and permeability coefficient (Kp) was observed in nanoemulsion formulations as compared to control or drug-loaded neat components. The highest value of these permeability parameters was obtained in optimized formulation B3, which consisted of 0.5% w/w of carvedilol, 6% w/w of oleic acid:IPM (1:1), 22.5% w/w of Tween 80, 22.5% w/w of Transcutol P, and 49% w/w of distilled water and in which the solubility of the drug was 4500-fold higher. The optimized nanoemulsion was characterized for pH, conductivity, viscosity, droplet size, droplet shape, and refractive index. Thermodynamic studies showed that there had been a significant decrease of 88% in activation energy (Eact) when the drug was incorporated in nanoemulsion. The irritation studies suggested that the optimized nanoemulsion was a non-irritant transdermal delivery system. PMID- 18402368 TI - Preparation and characterization of solid lipid nanoparticles loaded with alpha Asarone. AB - This work investigated the potential of solid lipid nanoparticles (SLNs) to improve oral bioavailablity and tissue uptake of a poorly soluble drug, alpha Asarone. Ultrasonic homogenization method was employed to prepare alpha-Asarone loaded SLNs (alpha-Asarone-SLNs). Particle size and distribution, pH, viscosity, drug incorporation and zeta potential of the SLNs were investigated. Pharmacokinetic study of oral administration to male rats at 10 mg/Kg suggested that the relative bioavailability of alpha-Asarone was significantly improved in alpha-Asarone-SLN group compared to alpha-Asarone solution group. Comparison of alpha-Asarone-SLN to alpha-Asarone control solution for alpha-Asarone concentrations in rat tissue showed an increased uptake of alpha-Asarone in brain and lung for the ARE-SLN group. These results indicate that alpha-Asarone-SLNs significantly enhance the absorption and tissue distribution of alpha-Asarone. SLNs offer a new approach to improve the oral bioavailability of poorly soluble drugs. PMID- 18402369 TI - Microemulsions as potential drug delivery systems: a review. AB - By many estimates, up to 40 percent of new chemical entities discovered by the pharmaceutical industry are poorly soluble or lipophilic compounds. Solubilization of hydrophobic drugs with low aqueous solubility has been a major area of interest in recent years. Various solubilization techniques involve the use of co-solvents and surfactants along with pH adjustments. Applications of microemulsions have also drawn attention in the field of solubilization techniques. Microemulsions are optically isotropic and thermodynamically stable systems consisting of water, oil, a surfactant, and a co-surfactant and are known to enhance the bioavailability of drugs via topical and systemic routes. The objective of this review is to present briefly the possible applications of these novel systems of microemulsions. Most studies reported in the literature have investigated microemulsions intended for dermatological application because of the wider range of potential excipients. PMID- 18402370 TI - Organizational unity. An important key to dentistry's success. PMID- 18402371 TI - No release, no refund. PMID- 18402372 TI - A hidden agenda. PMID- 18402373 TI - Managed care contracts need eyes of attorney, too. PMID- 18402374 TI - 2008 NYSDA President Stephen B. Gold. Caring, compassionate and a need to serve. PMID- 18402375 TI - Bisphosphonate-associated osteonecrosis. A clinician's reference to patient management. AB - Bisphosphonates, as inhibitors of osteoclasts, are widely used in the management of metastatic bone disease and in the prevention of osteomalacia and osteoporosis. Recent cases of bone necrosis of the jaws have been associated with the use of bisphosphonate therapy. A case is presented of a patient with osteonecrosis of the maxilla with a history of long-term bisphosphonate therapy for metastatic breast cancer. The authors treated the patient and suggest appropriate patient management guidelines with reference to current knowledge. Although a definitive treatment for bisphosphonate-associated osteonecrosis has not yet been established, clinicians must be aware of the pharmacologic properties of several bisphosphonates currently available and their indications, susceptible risk factors in the development of osteonecrosis of the jaws, the clinical signs and symptoms, and recommendations for patient management, including prevention and early recognition. PMID- 18402376 TI - Sequential development of multiple supplemental premolars. Four-year follow-up report. AB - Multiple supernumerary teeth are usually associated with various syndromes, but they are relatively uncommon as an isolated dental anomaly. The presence of multiple supplemental premolars in all four quadrants and especially in the maxilla is rare. This report presents a unique case of multiple supplemental premolars in all four quadrants that were distributed unevenly (two erupted; five unerupted). A series of radiographs taken over four years demonstrated further sequential developing bilateral supplemental premolars in the maxilla, where they are considered the rarest. Appropriate management and the importance of regular radiographic monitoring of such cases are briefly reviewed. PMID- 18402377 TI - The efficacy of two UV toothbrush sanitization devices. A pilot study. AB - The authors conducted a study to evaluate the efficacy of two different toothbrush sanitization machines for household use and to determine if one machine's sanitizing power was superior to the other. Bacteria from the same individual were tested for six days using the same brand toothbrush and toothpaste at the same time of day. The toothbrushes were sanitized using the VIOlight and HIGH DENT. In 83% of cases with the VIOlight machine, the amount of bacteria decreased after being sanitized, whereas the HIGH DENT had a 100% decrease in bacteria. Eighty-three percent of brushes sanitized with HIGH DENT resulted in rare or no growth compared to only 33% of those sanitized with VIOlight. While both machines reduced the amount of bacteria present on the toothbrush, the HIGH DENT was 50% more effective than VIOlight in reducing the majority of bacteria on the toothbrush. PMID- 18402378 TI - Oral health status of New Yorkers. PMID- 18402379 TI - Concrescence: report of rare complication. AB - Concrescence is an uncommon developmental anomaly that may influence exodontia as well as periodontal, endodontic, orthodontic and even prosthodontic diagnosis and treatment planning. Unexpected complications arising from this condition may lead to legal complications. To minimize risk and adverse outcome of treatment, consideration should be given to recognizing this condition and, possibly, modifying treatment planning. This article presents an unsuspected case of concrescence with a review of the literature. PMID- 18402380 TI - Why do I have white spots on my front teeth? AB - Aesthetics have always been important in mainstream society. Fluorosis, a condition of hypomineralization of the enamel, is at its most critical stage when a child is between 22 and 25 months of age. Because their swallowing reflex is not fully developed, children under the age of 6 can swallow between 25% and 33% of fluoridated toothpaste with each brushing. In order to better educate parents about fluorosis and its effect on children's teeth, it is worth revisiting the guidelines for toothpaste use. PMID- 18402381 TI - Sodium hypochlorite chemical burn. Case report. AB - Sodium hypochlorite (NaOCl) is a well documented and widely accepted intracanal irrigant used to disinfect the pulp space in endodontic therapy. NaOCl can be an extremely cytotoxic material; therefore, care must be taken with its use. The authors present a case of periradicular extravasation of NaOCl with sequelae of pain, swelling, ecchymosis and parasthesia consistent with a chemical burn. The patient was treated with a nonsurgical medical management approach. She experienced a complete resolution of signs and symptoms, aside from a persistent residual focal parasthesia. PMID- 18402382 TI - Drug utilization pattern in dental outpatients in tertiary care teaching hospital in western Nepal. AB - A high incidence of dental disease has been reported in Nepal. Previous studies, both in the Manipal Teaching Hospital, Pokhara, Nepal, and other centers revealed problems in the use of medicines in dentistry. A number of initiatives to improve prescribing have been carried out. The study presented here was undertaken to assess the impact of these initiatives on drug utilization among dental outpatients. The study was conducted among patients attending the dental outpatient department of the hospital over a six-month period. Demographic details were studied. The drug classes and individual drugs prescribed were recorded. The cost of drugs was calculateS using the outpatient pharmacy price list. The prescriptions were analyzed using the WHO/INRUD prescribing indicators. Anomalies were noted in prescribing. Improvement was noted in certain parameters compared to previous studies. The educational initiatives should be strengthened. Managerial interventions can be considered. Further studies are required. PMID- 18402383 TI - Expression of monocyte chemoattractant protein-1 in brain tissue of patients with intractable epilepsy. AB - OBJECTIVE: To investigate the expression ofmonocyte chemoattractant protein-1 (MCP-1) in the brain tissue of patients with intractable epilepsy (IE) and to explore its role in the pathogenesis of IE. MATERIAL: Patients with IE were collected from the Department of Neurosurgery of several Chinese hospitals between 2002 and 2005. The average age of these 40 patients was 23.65 +/- 10.14 (X +/- SD) years (11 - 58 years), with 19 males and 21 females. In addition to the typical symptoms and distinct EEG features of epilepsy, all recruited patients met the requirements of intractable epilepsy. METHODS: On the basis of positive results obtained from gene chip analysis, the expression of MCP-1 was first investigated in the brain tissue of IE patients (40 cases) using Western Blotting and immunohistochemistry and then compared with the controls. RESULTS: Gene chip scanning demonstrated that expression of MCP-1 mRNA was upregulated in the brain tissue of patients with IE. Western Blotting and immunohistochemical experiments further confirmed that, as compared with that in the control group, expression of MCP-1 protein was significantly increased in the IE patients (p < 0.05). CONCLUSION: These results suggest that MCP-1 are involved in the pathogenesis of IE. PMID- 18402384 TI - Early onset Alexander disease: a case report with evidence for manifestation of the disorder in neurohypophyseal pituicytes. AB - We report the first case of Alexander disease diagnosed and published in the region of former Czechoslovakia. The case was characterized by early (late infantile) onset, the absence of megacephaly but with extensive internal hydrocephaly, despite a patent aqueduct. Neuropathology revealed severe depletion ofoligodendroglia and myelin, loss of axons, prominent astrocytosis with massive intracellular, dense globular GFAP aggregates which differed from typical Rosenthal fibers. Additionally, many large aggregates of GFAP were located extracellularly. Globular GFAP aggregates were also identified in neurohypophyseal pituicytes. DNA analysis disclosed a heterozygous mutation c.1117G>A in the GFAP, which is predicted to lead to the amino acid exchange p.Glu-373Lys (E373K) in the C-terminal tail of the GFAP protein. The parents and a healthy sister did not show any variation in GFAP in somatic cells. PMID- 18402385 TI - Papillary glioneuronal tumor--contribution to a new tumor entity and literature review. AB - Papillary glioneuronal tumor (PGNT) is a recently identified low-grade mixed glial-neuronal neoplasm of juvenile and young adult patients. The WHO classification does not categorize this tumor as a separate entity, but rather considers it as a variant of ganglioglioma. We present a new case of this rare entity, representing the 3rd case of this lesion in Chinese patients and review the findings in 34 patients from different case reports found in the international literature. This report describes a histologically similar appearing lesion arising in the left frontoparietal lobe of a 23-year-old man. Its salient morphological characteristics are the presence of pseudopapillary structures composed of blood vessels, often hyalinized, lined by uniform small astrocytes and a proliferation of neurocytic cells which eventually admixed with ganglioid and ganglion cells. Sporadic Rosenthal fibers, foci of calcification, areas of hemosiderin deposition were identified. The mean Ki67 labeling index remained below 1%. Signs of anaplasia, in particular mitotic figures, endothelial proliferation or necrosis were consistently lacking. It is important that every new case of PGNT is reported to allow its recognition and classification. We perceive PGNT as a clinically and morphologically well-delineated subgroup of extraventricular low malignant potential neoplasm, whose presentation may allow for consideration as an entity. PMID- 18402386 TI - Congenital ependymoblastoma arising in the sacrococcygeal soft tissue: a case study. AB - Ependymoblastomas are distinct embryonal tumors of the central nervous system reported only rarely in the literature. Most examples arise in young children under the age of 2 years, in the supratentorial compartment, and may or may not be related to the ventricular system. We report the case of a one-day-old infant who presented with a 6.4 x 5.6 x 3.5 cm ruptured buttock mass. Ultrasound demonstrated a solid mass at the base of the spine that displaced the bladder anteriorly with resultant hydronephrosis. Magnetic resonance images confirmed the presence of a solid mass surrounding the lower sacrum with an internal component partially encircling and deviating the rectum. Histopathological evaluation confirmed the diagnosis of ependymoblastoma. Of note, immunohistochemical analysis revealed diffuse staining with vimentin and patchy expression of synaptophysin, glial fibrillary acidic protein, neurofilament proteins, neuron specific enolase, CD99 and nestin. On the 42nd day of life, chemotherapy was initiated with a modified Children's Oncology Group (COG) AGCT-01P1 (cyclophosphamide, cisplatin, 70% etoposide, no bleomycin) regimen. The authors describe their experience and review the literature, emphasizing that ependymoblastomas should be considered in the differential diagnosis of sacral masses in the newborn. PMID- 18402387 TI - DNA topoisomerase II-alpha as a proliferation marker in human gliomas: correlation with PCNA expression and patient survival. AB - OBJECTIVE: DNA topoisomerase II-alpha (Topo-IIalpha) is the inducible form of the enzyme responsible for the first step in the modification of DNA topology. Topo IIalpha upregulation has been demonstrated in different tumors. Topo-IIalpha products may modulate tumoral growth, metastasis and immunosuppression, inhibit apoptosis and cause resistance to chemotherapy. Moreover, the antitumoral effect ofTopo-IIalpha inhibitors has been documented. MATERIAL AND METHODS: We studied the immunohistochemical expression and the prognostic value of Topo-IIalpha on 57 surgical specimens ofglioma. Furthermore, we evaluated the correlation between the immunohistochemical expression of Topo-IIalpha and proliferating cell nuclear antigen (PCNA). RESULTS: A statistically significant correlation with survival time was found, there was no statistically significant difference in survival between patients receiving or not receiving carmustine-based combined chemotherapy (p > 0.05), regardless of histological type. A significant correlation between Topo-IIalpha and PCNA was documented (r = 0.9245, p < 0.001). CONCLUSIONS: Our findings show that gliomas immunohistochemically express Topo IIalpha that is correlated with PCNA expression, and which is significantly less frequent in long survivors. The presence of a statistical correlation with survival time and tumor histological grade encourages further studies on a larger series to verify the prognostic value of Topo-IIalpha expression in gliomas. PMID- 18402388 TI - Primary anaplastic ganglioglioma with a small-cell glioblastoma component. AB - Gangliogliomas usually present as benign tumors corresponding to World Health Organization (WHO) Grade I. Very rarely, gangliogliomas show histological features of malignancy and are then classified as anaplastic gangliogliomas of WHO Grade III or IV. In most cases, anaplastic gangliogliomas developed after radiation therapy or progression from a pre-existing low-grade ganglioglioma. Here, we report the case of a 77-year-old male patient who was operated on a primary ganglioglioma with a highly anaplastic glial component corresponding to a small-cell glioblastoma. To our knowledge, this is the first reported case of a primary anaplastic ganglioglioma with a small-cell glioblastoma component. PMID- 18402389 TI - Sarcoid neuropathy: clinico-pathological study of 4 new cases and review of the literature. AB - There are several reviews devoted to neurosarcoidosis and a few reports restricted to sarcoid neuropathy. Since 1989, we have investigated 4 new cases of sarcoid neuropathy, 1 with chronic sensory motor neuropathy (CSMN), another with painful neuropathy and 2 with atypical chronic inflammatory demyelinating polyneuropathy (CIDP). In each case, biopsy specimens from the superficial peroneal nerve and peroneus brevis muscle were taken by the same cutaneous incision and studied on paraffin sections, semi-thin sections and under electron microscope. We compared neuropathological findings from our 4 cases with those from 34 well-studied nerve biopsies previously reported in the literature, and which concerned 16 cases of CSMN, 13 cases ofmononeuropathy multiplex, 2 cases of painful neuropathy and three cases of CIDP. In all of these 38 cases of sarcoid neuropathy, the characteristic noncaseiting granulomas (NCG) were observed on the nerve in 11 cases, on the muscle alone in 5, on both muscle and nerve in 10, and in the nerve and another parenchyma in 4. In the 8 remaining cases, NCG were observed in another parenchyma, mainly lung or lymph nodes. Moreover, necrotizing vasculitis was present in nerve biopsies from 8 cases and microvasculitis without obvious necrosis in 2 others. Nerve fiber lesions, which are mainly axonal, are probably related to mechanical compression by NCG and/or to an ischemic process due to vasculitis. Cytokines and immune factors may also play a role, especially in certain cases with a clinical presentation of CIDP. PMID- 18402390 TI - Genetic testing for all forms of myotubular/centronuclear myopathy. PMID- 18402391 TI - [Tetralogy of Fallot: evolution of medical-surgical management (part 2)]. AB - This review is divided into two parts: the first of which traces the evolution of medical and palliative techniques; the following is a selective account of surgical concepts and procedures; the emphasis is on therapeutic developments during the last 50 years. The second part approaches the surgical corrections and tries to explain the reinterventional causes and the residual defects responsible of cardiac failure. PMID- 18402393 TI - [Primary vesico-ureteral reflux of the newborn]. AB - Vesico-ureteral reflux (VUR) and urinary tract infections (UTI) are more common in male than female infants. It was demonstrated that Voiding Detrusor Pressure (VDP) is higher in healthy males than healthy females and more in males with VUR versusmales without VUR. Baby boys with severe reflux may present also hypercontractility. During infancy, VDP and hypercontractility generally decrease: this is essential for reflux resolution. PMID- 18402392 TI - [Chronic sequelae in preterm infants]. PMID- 18402394 TI - Primary non-refluxing megaureter: need for conservative treatment. AB - PURPOSE: In the last two decades, many reports have confirmed the efficacy and safety of the conservative treatment of non-refluxing megaureter in asymptomatic patients and many cases of ureteral dilatation tend to resolve spontaneously. We report our experience on 108 patients with primary non-refluxing megaureter detected prenatally or diagnosed after birth and we discuss our results with long term non surgical treatment. MATERIAL AND METHODS: All patients were evaluated by ultrasound (US), voiding cystourethrogram (VCUG) and MAG3 renography. Observation period ranged from 6-72 months (mean 29.1). RESULTS: Surgery was performed in 12 patients (11.1%) with severe hydroureteronephrosis. Complete resolution or significant improvement was noted in 80 cases (74%) and persisted in 16 cases (14.8%). In the group with spontaneous resolution the ureteral diameter was less than in patients without resolution. Megaureters grade 1 to 3 tended to resolve between 12 and 36 months of observation. CONCLUSION: Conservative management is the treatment of choice in primary non refluxing megaureter. The grade of hydroureteronephrosis is an important predictor factor and infants should be followed periodically with renal ultrasound and diuretic renography. PMID- 18402395 TI - [Thoracoscopic treatment of oesophageal atresia]. AB - BACKGROUND: Advancements in minimally invasive surgery in newborns have allowed even the most complex neonatal procedures to be approached using these techniques. Other authors have demonstrated its efficacy in the treatment of the esophageal atresia with distal fistula. METHODS: We report our experience based on the thoracoscopic repair of esophageal atresia with distal fistula in two newborns. Birth weights were 2.800 g and 2.300 g respectively. The patients were intubated endotracheally and placed in a left prone position. Four trocars were inserted: the first one of 5 mm was positioned in the fifth intercostal space for the camera, the other two of 3 mm were positioned in the fourth intercostal space on the anterior and posterior axillary line respectively for the operative instruments. The last trocar of 3 mm was inserted in the third intercostal space on the anterior axillary line for the lung retractor. CO2 was insufflated at a pressure of 8 mm Hg and a flow of 0.5 L/min. The fistula was first isolated then ligated and cut with scissors. The proximal esophagus was opened and an anastomosis was made over a 6F or 8F nasogastric tube with interrupted 4-0 Vicryl sutures. A tube chest was placed through the lower trocar site with the tip near the anastomosis. RESULTS: These two procedures were free of neither intraoperative nor post-operative complications. Feeding by nasogastric tube was started after a mean of 4 days. Barium swallow made on day 7 demonstrated no leakage and no stenosis of the anastomosis. Total oral feeding was possible after 8 days. Mean hospitalization was 14 days. CONCLUSION: This initial report shows, as demonstrated by the experience since 1999 by other authors, that the thoracoscopic esophageal repair in the newborns is technically feasible and, thanks to a magnified vision, it allows to abtain a good isolation of the esophagus and of the tracheo-esophageal fistula respecting the anatomical structures. Moreover the advantages are in terms of exposure and esophageal length, avoiding the significant short and long-term morbidity associated with thoracotomy. PMID- 18402396 TI - [Gastric fibroid polyp in a 4-month-old girl with Costello syndrome]. AB - We describe the case of a 4-month-old girl with a gastric fibroid polyp. This was an occasional radiographic finding, confirmed by sonography and computerized tomography. This very rare benign tumor was surgically removed. The diagnosis of Costello syndrome was based on clinical appearance. This is the first report of a gastric fibroid polyp in Costello syndrome, a genetic disease with a high tumor frequency. PMID- 18402398 TI - Intussusception due to a cecal duplication cyst: a rare cause of acute abdomen. Case report. AB - Duplications of the alimentary tract are rare congenital anomalies. The ileum is the most common site, whereas rectal, duodenal, gastric and cecal duplications are extremely rare. Duplication cysts of the cecum, in a neonate, are even rarer, with only 19 cases reported in medical literature to date. We report a case of intestinal intussusception due to a cecal duplication cyst. PMID- 18402397 TI - [Pompe's disease: the role for early diagnosis and treatment]. AB - Pompe's disease (PD) is a glycogen storage disease characterized by the deposition of glycogen within body cells. This may lead to severe cardiac hypertrophy, with heart failure. The authors describe a female infant with PD, who developed severe cardiac hypertrophy, and was treated with recombinant human enzyme replacement therapy. This approach led to a progressive reduction of the heart hypertrophy, with improvement of the clinical condition. PMID- 18402400 TI - [Who wants to slough off the pediatric specialties?]. PMID- 18402399 TI - [How does the landscape change in lysosomal storage disease]. PMID- 18402401 TI - [Respiratory distress of the neonate and the rate of caesarean section have increased over the last 30 years. Is there a link?]. AB - In Switzerland, the rate of respiratory distress in neonates needing hospitalization has doubled over the last thirty years, concerning in particular babies weighing more than 2500 g. In the same time, the rate of Caesarean section (CS) has also multiplied. We suppose that a link between the two evolutions might be the increase of elective CS. They tend to be planned early at term to avoid the onset of spontaneous labour As a consequence, the foetus is deprived of different mechanisms helping pulmonary transition around birth. The potential benefits of CS regarding morbidity of foetus and mother should not overshadow that CS is a significant risk factor for respiratory problems of the neonate. This risk could be dramatically decreased by planning elective CS only after completed 39 weeks of gestation. PMID- 18402402 TI - [Intrauterine growth restriction: impact on brain development and function]. AB - Evidence exists that the developing organism adapts to the environment it finds itself. Short and long-term adjustments take place and will initially induce intrauterine growth retardation but will also have consequences that will appear later in life. These adjustments are referred as "programming". The use of advanced magnetic resonance imaging techniques in IUGR babies has delineated changes in the development of the central nervous system that correlate with altered neurodevelopment and could be implicated in the development of neuropsychiatric disorders in adult life. In this review, we will delineate some modifications of CNS development and functions that occur after exposition to adverse environment and that can now be studied in vivo with advanced imaging technology. PMID- 18402403 TI - [How to investigate sleep related breathing disorders in children?]. AB - Sleep related breathing disorders (SDB) in children has a major negative impact on their neurocognitive development and should be identified and treated early as to decrease morbidity. Children do not usually present daytime fatigue and sleepiness like adults, but rather show abnormal behavior patterns and learning disabilities. The presence of sustained nightime snoring is a good screening toll for SDB. Polysomnography is the gold standard for diagnosis even though nightime oxymetry, with its high positive predictive value, can also be used. The most frequent SDB in childhood is sleep obstructive apnea syndrome (SOAS), which generally requires surgical treatment with adenotonsillectomy. Long term evolution of pediatric SOAS still has to be defined, as the role of other diagnostic tools such as outpatient polygraphy. PMID- 18402404 TI - [Migraine and symptomatic headache in children]. AB - Pediatric migraine differs from adult migraine especially in regards to duration, localisation and quality of pain. A detailed description of the symptoms with a normal neurological examination allows in most cases to rule out secondary headaches without other exams. Many different medications are used for symptomatic or prophylactic treatment with success. Symptomatic headaches should be suspected if there is any abnormality in history or neurological exam. Headaches due to arterial hypertension, ENT problems or maxillofacial causes should not be forgotten. Intracranial hypertension should be excluded especially in children with ventriculo-peritoneal shunt, since shunt dysfunction can be fatal. Post traumatic headache can be impressive but have a good prognosis. PMID- 18402405 TI - [Periodic fever in children: keep in mind the PFAPA syndrome]. AB - The autoinflammatory diseases should be considered in the differential diagnosis of recurrent fever in childhood. These diseases are characterized by inflammatory episodes without an evident cause. Some of these diseases, like the Familial Mediterranean Fever, have a genetic origin and need a chronic treatment to avoid severe complications on the long term. PFAPA syndrome is the most frequent cause of recurrent fever and is diagnosed based on unspecific criteria. The treatment is still controversial. One dose of Prednisone is able to interrupt the flare and tonsillectomy may induce a remission in the majority of the cases. PMID- 18402406 TI - [How to treat childhood obesity? Importance of primary prevention]. AB - The prevalence of childhood obesity increases dramatically. First signs of cardiovascular diseases and type 2 diabetes appear early in life. The treatment of childhood obesity aims at weight maintenance during growth, normalization of body mass index at long-term and prevention of complications. The family based behavioural therapy is a promising approach. It provides simultaneous treatment for the overweight parent and child in order to modify the family environment, to provide role models and support for child behaviour changes. However, this requires group leaders and multiple counselors to meet with families. The treatment should be initiated as soon as possible, as its efficacy is reduced after the onset of puberty. Early preventive interventions that aim to modify both individual's behaviours and the environment are needed. PMID- 18402407 TI - [Drugs and breastfeeding: some facts to consider before deciding they are incompatible]. AB - Breastfeeding is considered the ideal way of providing young infants with the required nutrients for healthy growth and development. An increase in the prevalence and duration of breastfeeding has been observed over the last years in response to promotion campaigns. When a medicamentous treatment is started, discontinuation of breastfeeding is often proposed, for fear of harmful consequences for the infant. Nevertheless such a decision is not unimportant, and it appears that many drugs can actually be used during breastfeeding without significant risk. An assessment of the real risk incurred by exposed children makes it possible to avoid unnecessary discontinuations of breastfeeding. This article aims to review the facts needed to assess amount of drug exposure to the child and to list the few drugs associated with significant effects on the nursing infant. PMID- 18402408 TI - [Anatomophysiological consideration on the sacral reflex]. AB - Sacral reflex testing is a very specific one. Coming from sensory receptors afferent neuronal pudendal passway enters the spinal cord S2 to S4 via spinal dorsal roots and transmits the bioelectrical activity to a complex interneuronal circuitry. Information is processed at this level and dispached into spinal motoneurones for innervating the pelviperineal muscles according to their specific radicular origins. Selective staged sacral reflexes contribute to locate the site of compression. Sacral reflex configuration was based on clinical and electrophysiological findings which is actually confirmed and valided by anatomical studies. This testing is the most efficacious and useful procedure of assessing the pudendal neuralgia, the S2 to S4 reflexes and the pelvic floor dysfunctions. PMID- 18402409 TI - [Management problems in the ECG of an elderly person. Case no. 9]. PMID- 18402410 TI - [Breast cancer: recommendations for its prevention (I)]. PMID- 18402411 TI - [Is it necessary to treat subclinical thyroid diseases?]. PMID- 18402412 TI - [The archbishop of Paris and the status of the human embryo (I)]. PMID- 18402413 TI - [Of ecology and virginity(2)]. PMID- 18402414 TI - [Breathing under a dome]. PMID- 18402415 TI - Women's journey of recovery from sexual assault trauma: a grounded theory--part 1. AB - Thousands of women and children experience sexual assault trauma annually in South Africa. The challenge posed by recovery from sexual assault trauma is a reality that confronts the survivors of sexual assault, their families and the larger community of service providers. Yet, little research has been conducted on recovery from sexual assault as a phenomenon. The purpose of the study was to explore and analyse the journey of recovery which is undertaken by women who have been sexually assaulted, with the aim of discovering and developing the grounded theory of recovery from sexual assault trauma within the first six months following the event of rape. The main research question was: What constitutes the journey of recovery undertaken by women within the first six months following sexual assault? A longitudinal qualitative study was conducted using the principles of grounded theory methodology as proposed by Strauss and Corbin (1990, 1998). A series of in-depth one-to-one interviews were conducted with a sample of ten women. The participants were selected through open, purposive and theoretical sampling procedures. The study was conducted over a period of six months following the event of sexual assault. The substantive theory was discovered and constructed through the inductive and deductive analysis of data, grounded on the ten women's descriptions of their journey of recovery from sexual assault. The theory of women's journey of recovery that was discovered and developed consisted of eight theoretical concepts or categories. These included the following concepts: 1. Sexual assault trauma 2. Awakening 3. Pragmatic acceptance 4. Turning point 5. Reclaiming what was lost 6. Defining own landmarks of healing 7. Readiness for closure 8. Returning to self. The grounded theory of the journey of recovery from sexual assault is a contribution to the knowledge about women's journey of recovery from sexual assault. It provides a process and language for understanding women's experiences within the larger social context of recovery from assault. The findings suggested that women's journey of recovery from sexual assault trauma is an individual experience that is influenced by many and varied causal and intervening conditions. These included personal biographies, relationships, supportive and non-supportive environments at individual, family, community and societal levels. Recommendations for future research and clinical management of sexual assault are made based on the researcher's interpretation of the findings. PMID- 18402416 TI - Women's journey of recovery from sexual assault trauma: a grounded theory--part 2. AB - The purpose of the study was to explore and analyse the journey of recovery which is undertaken by women who have been sexually assaulted, with the aim of discovering the grounded theory of recovery from sexual assault within the first six months following the event of rape. The main research question was: 'What is the journey of recovery that is undertaken by women within the first six months following sexual assault?' Another question that developed during data collection and data analysis was 'What is the meaning that women attach to recovery?' The findings are discussed under the eight concepts or categories and the context and the intervening conditions that influence the journey of recovery from sexual assault trauma. Refer to part 1 article. These are complemented with abstracts of data from the participants' voices and the related discussions. The developed theory highlights the process and the interconnectedness of the different stages of what the women experience in their journey of recovery from sexual assault trauma. PMID- 18402417 TI - Exploration of knowledge, attitudes and percepions regarding sexually transmitted infections among patients attending a primary health care clinic in Gauteng--part 1. AB - Sexually transmitted infections are a major public health problem in South Africa. The high incidence and prevalence of sexually transmitted infections in South Africa pose a serious threat to public health for two main reasons. Firstly, the long-term consequences of these infections are a major cause of loss of health or life, and secondly, sexually transmitted infections are important co factors in driving the human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) epidemic. The control and prevention of sexually transmitted infections has become an urgent priority. At a primary health care clinic, the nursing personnel experienced an increasing number of persons daily seeking treatment for sexually transmitted infections. The purpose of the study was therefore to investigate the knowledge, attitudes and perceptions of people seeking treatment at Stanza Bopape Clinic regarding sexually transmitted infections. The study was a contextual, quantitative survey. The population was all patients (15 years and older) who visited the clinic for a health-related problem during November 2004 to January 2005. The sampling method was convenient. The data gathering method was self-report using a structured questionnaire basic on current knowledge. The data analysis was done with descriptive statistics. PMID- 18402418 TI - Guidelines to reduce sexually transmitted infections at primary health care centres--part 2. AB - Sexually transmitted diseases are the second highest cause of death among women aged 15-44 years. The incidence and prevalence of sexually transmitted diseases is on the increase at many health care centres despite their dreadfulness and many preventative measures. According to the Scope of Practice of registered professional nurses working in a community health clinic, the registered professional nurse has a responsibility to prevent disease and promote health as well as do research. This paper aims to describe guidelines to reduce sexually transmitted infections at primary health care centres based on research done to investigate the community specific knowledge, attitudes and perceptions regarding sexually transmitted infections. Structured interviews were used to collect data from 1694 participants (males and females) who were consulted at a primary health care clinic in Mamelodi, Gauteng province. Existing patients' records were also utilised to calculate the incidence rate of sexually transmitted infections. Data were computerised and analysed quantitatively to reflect the results in frequencies and percentages as reported in Part 1 of this research. Guidelines to direct the practice of registered professional nurse were developed based on the findings of the results. These guidelines must underpin community specific interventions to reduce the incidence of sexually. PMID- 18402419 TI - Knowledge of breast self-examination in women in Sierra Leone. AB - This research is a follow up of a Breast Week which was organized in Freetown, Sierra Leone. The specific objective of this study was to assess the effectiveness of the knowledge and teachings given to the women who participated in this project. A quantitative approach with an exploratory descriptive design was adopted and an observational checklist guided the data collection process. A sample size of 120 women (10%) who participated in the Breast Week was obtained through systematic sampling. During the Breast Week women were taught how to examine their breasts using breast self-examination (BSE) to detect abnormalities of the breasts. This study was undertaken one year later by the researcher to ascertain whether the information on breast self-examination provided during the Breast Week was being utilised and whether what was taught was being put into use. Data were analyzed using SPSS version 11.5. Reliability and validity were ensured through the use of a structured observational checklist and a pilot study was undertaken. The observations were all observed and recorded by the same researcher. The majority of the 120 women (91.7%) stated that they had never practiced BSE before the Breast Week. After receiving health education on BSE, 95% were able to demonstrate an effective method of undertaking BSE. It is thus recommended that every opportunity should be utilized in health care settings to teach BSE and to reinforce the practice, especially in poverty stricken countries where other forms of screening methods are unavailable. PMID- 18402420 TI - A concept analysis of holistic nursing care in paediatric nursing. AB - Holistic nursing care is widely advocated and is espoused in the philosophy of the South African Nursing Council. This concept is unclear, variously interpreted and poorly understood in paediatric nursing. This study was undertaken to examine the meaning of holistic nursing care and to develop a framework for holistic nursing care, which can be utilised in nurse education settings and in clinical nursing practice in the context of paediatric nursing. A qualitative, interpretive, explorative and contextual research design was used. An evolutionary concept analysis was undertaken to clarify the concept "holistic nursing care" in paediatric nursing in three Johannesburg hospitals. Rodgers' (1989, 2000) evolutionary method was utilised to analyse the concept. The study objectives were formulated in two phases to: --Conduct an analysis of the concept "holistic nursing care" --Obtain an emic viewpoint of holistic nursing care from paediatric nurses working in the academic hospitals. --Identify the characteristics and dimensions of "holistic nursing care" and develop a framework of holistic nursing care for paediatric nurses working in the academic hospitals. Attributes of holistic nursing care yielded two dimensions; whole person and mind body-Spirit dimension. The decriptors of whole-person include physical, mental, emotional, spirit and spitual being. Spirituality is the predominant antecedent. Holistic nursing care is initiated by the recognition of the individual as a spiritual being with a mind-body-spirit dimension. Spirituality is an ever present force pervading all human experience. Complimentary alternative medicine (CAM) was identified as a surrogate term. The connection of CAM with holistic nursing care is the focus of therapeutic interventions that are directed to the mind-body-spirit dimension. Therapeutic interventions are designed to meet the needs of the whole-person. Caution is advocated in the use of CAM therapies in child nursing, as CAM efficacy has not been sufficiently investigated in child health care. PMID- 18402421 TI - Introducing human rights and health into a nursing curriculum. AB - AIM: The introduction of a module in Health and Human Rights into a Postgraduate diploma curriculum for registered nurses is described. BACKGROUND: An important component of nursing programmes in South Africa has been teaching of the principles of ethical practice and relevant ethical codes. A number of factors have contributed to the need to include human rights as an integral component of nursing curricula in South Africa. These include the Bill of Rights in the Constitution of South Africa and the implications thereof for health care delivery, the primary health care approach in the delivery of health care in South Africa, the development and acceptance of Patients' Rights Charters, and the recognition of the role that health professionals played--whether through lack of knowledge and awareness or direct involvement--in the human rights violations in the health sector exposed during the hearings of the Truth and Reconciliation Commission. MODULE DESCRIPTION: An outline of themes covered in the course content is described, together with examples of learning activities and teaching materials. Reflections of the course convenor and students are used to highlight the importance of inclusion of health and human rights as foundational to ethical nursing practice, regardless of the health care setting. PMID- 18402422 TI - Rationalisation of nursing education in Limpopo province : nurse educators' perspectives. AB - Nursing education institutions are facing a challenge of realigning its functioning according to the changes that are taking place within the country. The intention of the government post apartheid was to correct the imbalances which were brought about by the apartheid government and the following regulations and policies influenced the change in nursing education, that is, Reconstruction and Development Programme (RDP), White Paper on Higher Education (WPHE), and the National Qualification Framework (NQF) (South Africa, 1995:6). In 1996 the government introduced the first democratic constitution of the Republic of South Africa (RSA) according toAct 108 of 1996. In the light of those increasing changes in nursing education, led by political change, the experiences of nurse educators is a critical issue facing nursing campuses. The purpose of this study was two-fold; namely: to explore and describe the experiences of nurse educators with regard to the rationalisation of nursing education and to use information obtained to describe guidelines for the effective rationalisation of a nursing college in the Limpopo Province. A qualitative, exploratory, descriptive and contextual research design was used. Qualitative interviews were conducted with nurse educators who worked in nursing colleges before and after 1994. Measures to ensure trustworthiness were applied and ethical issues were adhered to throughout the research process. Data was analysed following Tesch's method (Creswell 1994:154-155). The research established that nurse educators experienced dissatisfaction in several areas relating to the rationalization of nursing education. Support was also expected from bureaucracy at higher level. This study developed guidelines to policy makers and nurse educators to ensure effective rationalisation process. PMID- 18402423 TI - Plain language for interpreting in consulting rooms. AB - Intercultural communication is by nature a complex activity. In a multilingual society like ours, it will inevitably surface in the health care sector. The services of an interpreter are often considered to break the impasse in this communication process. The communication problem between the two parties, the service provider and client/patient, is often not simply a matter of language but societal factors of which the liaison interpreter should be aware of also plays a major role for effective extended communication. This article focuses on some of the problems in rendering an oral source text in multilingual and multicultural societies such as South Africa in which there are heterogeneous target audiences for interpreting. It is pointed out that interpreters in such societies must take into account the heterogeneity of the target audiences, or otherwise interpreting will only be symbolic gestures, empty of value, and thus not communicate the message intended. In the process the limitations of the interpreter and how the presence of the interpreter can be facilitated, is also highlighted. PMID- 18402424 TI - Risk factors for cardiovascular disease in the Ga-Rankuwa community. AB - Cardiovascular disease is the most common and yet one of the most preventable causes of death in the world. Rapid urbanization in South Africa is accompanied by rapid changes in lifestyle and environmental exposure that increase the burden of chronic cardiovascular diseases. Risk factors, modifiable or nonmodifiable, exist that increases a person's chances of developing cardiovascular disease. Though some knowledge is available about the prevalence of the risk factors in South Africa, no information is available regarding the community of Ga-Rankuwa. The purpose of the study was therefore to investigate the prevalence of risk factors for cardiovascular disease amongst the working-age people (18-40 years) in Ga-Rankuwa community. A quantitative survey was done and the sample was selected from zone 1, 2, 4, and 16 of Ga-Rankuwa from July 2005 to October 2005. The sampling method was census sampling (n=604). The data-gathering was self report using a structured questionnaire as well as physical measurement. Data were analysed using descriptive statistics. The results indicated that risk factors, specifically obesity, physical inactivity and hypertension, were very prevalent in Ga-Rankuwa community. Different distributions of risk factors exist in the various sex and age groups. This finding again emphasises the importance of not developing health interventions with a single focus, for example hypertension or obesity. The risk factors are interwoven and affect each other. It is important to initiate a comprehensive health project to lower the risk factors of cardiovascular diease in the Ga-Rankuwa community. PMID- 18402425 TI - [Diseases of the kidney tubules]. PMID- 18402426 TI - [Acute and chronic pancreatitis--an overview]. PMID- 18402427 TI - [Acute coronary syndrome]. PMID- 18402428 TI - [Pulmonary hypertension]. PMID- 18402429 TI - Computational study on the aminolysis of beta-hydroxy-alpha,beta-unsaturated ester via the favorable path including the formation of alpha-oxo ketene intermediate. AB - The possible mechanisms of the aminolysis of N-methyl-3-(methoxycarbonyl)-4 hydroxy-2-pyridone (beta-hydroxy-alpha,beta-unsaturated ester) with dimethylamine are investigated at the hybrid density functional theory B3LYP/6-31G(d,p) level in the gas phase. Single-point computations at the B3LYP/6-311++G(d,p) and the Becke88-Becke95 1-parameter model BB1K/6-311++G(d,p) levels are performed for more precise energy predictions. Solvent effects are also assessed by single point calculations at the integral equation formalism polarized continuum model IEFPCM-B3LYP/6-311++G(d,p) and IEFPCM-BB1K/6-311++G(d,p) levels on the gas-phase optimized geometries. Three possible pathways, the concerted pathway (path A), the stepwise pathway involving tetrahedral intermediates (path B), and the stepwise pathway via alpha-oxo ketene intermediate due to the participation of beta-hydroxy (path C), are taken into account for the title reaction. Moreover, path C includes two sequential processes. The first process is to generate alpha oxo ketene intermediate via the decomposition of N-methyl-3-(methoxycarbonyl)-4 hydroxy-2-pyridone; the second process is the addition of dimethylamine to alpha oxo ketene intermediate. Our results indicate that path C is more favorable than paths A and B both in the gas phase and in solvent (heptane). In path C, the first process is the rate-determining step, and the second process is revealed to be a [4+2] pseudopericyclic reaction without the energy barrier. Being independent of the concentration of amine, the first process obeys the first order rate law. PMID- 18402430 TI - Theoretical elucidation of conflicting experimental data on vertical ionization potentials of microhydrated thymine. AB - In a recent article we reported calculations of the ionization energy thresholds (IET) of microhydrated thymine (Close; et al. J. Phys. Chem. A, 2006, 110, 7485). Calculations showed a distinct effect of microhydration on the IET's of thymine. The first water molecule was seen to decrease the IET by about 0.1 eV, and the second and third water molecules caused a further decrease of less than 0.1 eV each. These changes in IET calculated for the canonical form of thymine with 1-3 waters of hydration are smaller than the experimental values determined by Kim et al. (J. Phys. Chem. C 1996, 100, 7933). In the present study it has been shown that there is considerable reorientation of the water molecules in microhydrated thymine upon ionization. This leads to the expectation that the experimental ionization energies may therefore represent an adiabatic process. The results presented here show that the changes in experimental ionization energies determined by Kim et al. for microhydrated thymine are in good agreement with the calculated adiabatic ionization energies. PMID- 18402431 TI - Azo-hydrazone tautomerism in protonated aminoazobenzenes: resonance Raman spectroscopy and quantum-chemical calculations. AB - The protonation effect on the vibrational and electronic spectra of 4 aminoazobenzene and 4-(dimethylamino)azobenzene was investigated by resonance Raman spectroscopy, and the results were discussed on the basis of quantum chemical calculations. Although this class of molecular systems has been investigated in the past concerning the azo-hydrazone tautomerism, the present work is the first to use CASSCF/CASPT2 calculations to unveil the structure of both tautomers as well the nature of the molecular orbitals involved in chromophoric moieties responsible for the resonance Raman enhancement patterns. More specifically both the resonance Raman and theoretical results show clearly that in the neutral species, the charge transfer transition involves mainly the azo moiety, whereas in the protonated forms there is a great difference, depending on the tautomer. In fact, for the azo tautomer the transition is similar to that observed in the corresponding neutral species, whereas in the hydrazone tautomer such a transition is much more delocalized due to the contribution of the quinoid structure. The characterization of protonated species and the understanding of the tautomerization mechanism are crucial for controlling molecular properties depending on the polarity and pH of the medium. PMID- 18402434 TI - Distributed chemical computing using ChemStar: an open source java remote method invocation architecture applied to large scale molecular data from PubChem. AB - We present the application of a Java remote method invocation (RMI) based open source architecture to distributed chemical computing. This architecture was previously employed for distributed data harvesting of chemical information from the Internet via the Google application programming interface (API; ChemXtreme). Due to its open source character and its flexibility, the underlying server/client framework can be quickly adopted to virtually every computational task that can be parallelized. Here, we present the server/client communication framework as well as an application to distributed computing of chemical properties on a large scale (currently the size of PubChem; about 18 million compounds), using both the Marvin toolkit as well as the open source JOELib package. As an application, for this set of compounds, the agreement of log P and TPSA between the packages was compared. Outliers were found to be mostly non druglike compounds and differences could usually be explained by differences in the underlying algorithms. ChemStar is the first open source distributed chemical computing environment built on Java RMI, which is also easily adaptable to user demands due to its "plug-in architecture". The complete source codes as well as calculated properties along with links to PubChem resources are available on the Internet via a graphical user interface at http://moltable.ncl.res.in/chemstar/. PMID- 18402432 TI - Structural determinants for affinity enhancement of a dual antagonist peptide entry inhibitor of human immunodeficiency virus type-1. AB - Structure-activity correlations were investigated for substituted peptide conjugates that function as dual receptor site antagonists of HIV-1 gp120. A series of peptide conjugates were constructed via click reaction of both aryl and alkyl acetylenes with an internally incorporated azidoproline 6 derived from the parent peptide 1 (12p1, RINNIPWSEAMM). Compared to 1, many of these conjugates were found to exhibit several orders of magnitude increase in both affinity for HIV-1 gp120 and inhibition potencies at both the CD4 and coreceptor binding sites of gp120. We sought to determine structural factors in the added triazole grouping responsible for the increased binding affinity and antiviral activity of the dual inhibitor conjugates. We measured peptide conjugate potencies in both kinetic and cell infection assays. High affinity was sterically specific, being exhibited by the cis- but not the trans-triazole. The results demonstrate that aromatic, hydrophobic, and steric features in the residue 6 side-chain are important for increased affinity and inhibition. Optimizing these features provides a basis for developing gp120 dual inhibitors into peptidomimetic and increasingly smaller molecular weight entry antagonist leads. PMID- 18402435 TI - Indirect similarity based methods for effective scaffold-hopping in chemical compounds. AB - Methods that can screen large databases to retrieve a structurally diverse set of compounds with desirable bioactivity properties are critical in the drug discovery and development process. This paper presents a set of such methods that are designed to find compounds that are structurally different to a certain query compound while retaining its bioactivity properties (scaffold hops). These methods utilize various indirect ways of measuring the similarity between the query and a compound that take into account additional information beyond their structure-based similarities. The set of techniques that are presented capture these indirect similarities using approaches based on analyzing the similarity network formed by the query and the database compounds. Experimental evaluation shows that most of these methods substantially outperform previously developed approaches both in terms of their ability to identify structurally diverse active compounds as well as active compounds in general. PMID- 18402436 TI - Reporting a new class of divanadium(V) compounds connected by an unsupported hydroxo bridge. AB - Dinuclear oxovanadium(V) compounds [LV(V)O(mu-OH)OV (V)L](PF6) [H2L = N,N'-tert ethylene bis(salicylideneimine) (H 2Salen) and its derivatives] ( 1- 3) have been obtained by aerial oxidation of V (IV)OL precursors in THF in the presence of added NH 4PF 6. The oxidized vanadium(V) probably extracts an OH (-) ligand from the residual moisture in the solvent and is retained as an unsupported hydroxo bridge between the metal centers of these compounds as confirmed by single crystal X-ray diffraction analyses. The molecules of 1- 3 have centrosymmetric structures with each vanadium center having a distorted octahedral geometry. The bridging OH (-) group is located trans to the terminal VO t bond. The latter exerts strong trans labilizing influence to set the participating vanadium centers apart by about 4.1 A. These separations are by far the largest (e.g., V...V#, 4.131 A in 1) among all binuclear compounds containing an unsupported hydroxo bridge reported to date. The complexes retain their identity also in solution as established by (1)H NMR spectroscopy. Electrochemically, the behaviors of 1-3 are quite interesting as studied by cyclic voltammetry in acetonitrile, each undergoing three (except 3) nearly reversible metal-based reductions, all in the positive potential range (e.g., at E (1/2) = 0.57, 0.39, and 0.04 V versus Ag/AgCl reference for 1) as indicated by steady state voltammetry. The electrode process at 0.39 V appears to involve a single-step two electron transfer as revealed from the normal and differential pulse voltammetric data and probably includes a combination of V(V)-V(IV) <--> V(III)-V(IV) mixed oxidation states. Compounds 1-3 thus provide a unique example of divanadium compounds in which the metal centers are linked by an unsupported hydroxo-bridge. PMID- 18402437 TI - Self-assembly of an azido-bridged [NiII6] cluster featuring four fused defective cubanes. AB - The cluster [Ni6(H2L)2(HL1)2(N3)8].2C2H5OH.2H2O [1.2C2H5OH.2H2O], featuring four fused defective cubanes, has been obtained via azido-bridge-driven dimerization of two phenolate-centered trinuclear Ni3 fragments. PMID- 18402438 TI - Structure, cytotoxicity, and DNA-cleavage properties of the complex [Cu(II)(pbt)Br2]. AB - The reactions of the ligand 2-(2-pyridyl)benzthiazole (pbt) with CuBr 2 and ZnCl 2 in acetonitrile produce the complexes [Cu(pbt)Br 2] ( 1) and [Zn(pbt)Cl 2] ( 3), respectively. When complex 1 is dissolved in DMF, complex 2 is obtained as light-green crystals. The reaction of pbt with CuBr 2 in DMF also yields the complex [Cu(pbt)Br 2(dmf)] ( 2) (dmf = dimethylformamide). Complexes 1- 3 were characterized by X-ray crystallography. Complexes 1 and 3 have distorted tetrahedral coordination environments, and complex 2 is constituted of two slightly different copper centers, both exhibiting distorted trigonal bipyramidal geometries. Complexes 1 and 2 cleave phiX174 phage DNA, both in the presence and the absence of reductant. The free ligand pbt does not show any DNA-cleaving abilities. The poor solubility of complex 3 makes it not applicable for biological tests. The occurrence of DNA breaks in the presence of various radical scavengers suggests that no diffusible radicals are involved in the DNA cleavage by complex 1, as none of the scavengers inhibit the cleavage reaction. The DNA cleavage products are not religated with the enzyme T4 DNA ligase, which is an additional proof that the cleavage is nonhydrolytic. Most probably the cleaving reaction involves reactive oxygen species, which could not be trapped, leading to an oxidative mechanism. An easy oxidation of Cu (II)(pbt)Br 2 to Cu (III) in DMF and the reduction of the same to Cu (I), under similar electrochemical conditions may lead to the in situ activation of molecular oxygen, resulting in the formation of metal solvated nondiffusible radicals able to prompt the oxidative cleavage of DNA. Complex 1 and the pure ligand exhibit remarkable cytotoxic effects against the cancer cell lines L1210 and A2780 and also against the corresponding cisplatin-resistant mutants of these cell lines. PMID- 18402439 TI - A versatile series of nickel(II) complexes derived from tetradentate imine/pyridyl ligands and various pseudohalides: azide and cyanate compared. AB - The chemical reactions of a family of tetradentate pyridyl/imine ligands, L1, L2, and L3 (L1=[ N, N'-bis(2-pyridinylmethylene)]ethane-1,2-diamine; L2=[ N, N' bis(pyridin-2-yl)benzylidene]ethane-1,2-diamine; L3=[ N, N'-bis(2 pyridinylmethylene)]propane-1,3-diamine), with Ni (II) in the presence of various pseudohalides (N3(-), SCN(-), and NCO(-)) have served to prepare six different complexes, [Ni 2(L1)2(N3)2](ClO4)2.H2O (1), [Ni 2(L2)2(N3)2](ClO4)2 (2), [Ni2(L2)2(NCS)4] (3), [Ni2(L2)2(NCO) 2](ClO4)2 (4), [Ni2(L3)2(NCO)2](ClO4)2 (5), and [Ni(L3)(N 3)2] (6), which have been characterized by X-ray crystallography. Interestingly, four of these complexes are dinuclear and exhibit end-on (EO) pseudohalide bridges (1, 2, 4, and 5), one is dinuclear and bridged exclusively by the tetradentate ligand (3), and one is mononuclear (6). The bulk magnetization of the complexes bridged by EO pseudohalides has been studied, revealing these ligands to mediate ferromagnetic coupling between the Ni(II) ions, with modeled coupling constants, J, of +31.62 (1), +28.42 (2), +2.81 (4), and +1.72 (5) cm(-1) (where the convention H=-2JS1S2 was used). The striking difference in the coupling intensity between N3(-) and NCO(-) has prompted an investigation by means of density functional theory calculations, which has confirmed the experimental results and provided insight into the reasons for this observation. PMID- 18402440 TI - Steric influence on the excited-state lifetimes of ruthenium complexes with bipyridyl-alkanylene-pyridyl ligands. AB - The structural effect on the metal-to-ligand charge transfer (MLCT) excited-state lifetime has been investigated in bis-tridentate Ru(II)-polypyridyl complexes based on the terpyridine-like ligands [6-(2,2'-bipyridyl)](2-pyridyl)methane ( 1) and 2-[6-(2,2'-bipyridyl)]-2-(2-pyridyl)propane ( 2). A homoleptic ([Ru( 2) 2] (2+)) and a heteroleptic complex ([Ru(ttpy)( 2)] (2+)) based on the new ligand 2 have been prepared and their photophysical and structural properties studied experimentally and theoretically and compared to the results for the previously reported [Ru( 1) 2] (2+). The excited-state lifetime of the homoleptic Ru (II) complex with the isopropylene-bridged ligand 2 was found to be 50 times shorter than that of the corresponding homoleptic Ru (II) complex of ligand 1, containing a methylene bridge. A comparison of the ground-state geometries of the two homoleptic complexes shows that steric interactions involving the isopropylene bridges make the coordination to the central Ru (II) ion less octahedral in [Ru( 2) 2] (2+) than in [Ru( 1) 2] (2+). Calculations indicate that the structural differences in these complexes influence their ligand field splittings as well as the relative stabilities of the triplet metal-to-ligand charge transfer ( (3)MLCT) and metal-centered ( (3)MC) excited states. The large difference in measured excited-state lifetimes for the two homoleptic Ru (II) complexes is attributed to a strong influence of steric interactions on the ligand field strength, which in turn affects the activation barriers for thermal conversion from (3)MLCT states to short-lived (3)MC states. PMID- 18402441 TI - Tailoring the key in a molecular lock-and-key model system: the propylene oxide...2-fluoroethanol complex. AB - The conformational isomerism of the propylene oxide (PO)...2-fluoroethanol (FE) complex has been investigated using molecular beam Fourier-transform microwave spectroscopy complemented with high level ab initio calculations. Rotational transitions of three different binary conformers have been observed experimentally. On the basis of the agreement of the experimental and calculated rotational constants, they could be identified as the three most stable structures, anti G-g+, anti G+g-, and syn G+g-. All the observed structures exhibit a primary O-H...O hydrogen bond, an intramolecular O-H...F hydrogen bond and two secondary intermolecular C-H...F contacts. The two anti conformers, with FE and the PO methyl group on the opposite sides of the oxirane ring, show higher abundances than the syn conformer. In all three observed conformers, FE remains approximately in its favorable compact gauche conformation. The monofluorination of the molecular lock-and-key model system PO...ethanol increases not only the number of possible binary conformers, but also the discrimination energy among them. The superior discrimination ability of FE as compared to ethanol classifies it as a tailored key to the PO lock. PMID- 18402442 TI - Supramolecular control for the modular synthesis of pseudopeptidic macrocycles through an anion-templated reaction. AB - The anion-templated synthesis of different pseudopeptidic macrocycles has been studied in detail by using a multidisciplinary approach. The reaction between an open-chain pseudopeptidic diamine and the appropriate dialdehyde is highly affected by the presence of the best fitting anionic template. The formation of the corresponding macrocyclic tetraimino-template supramolecular complex is demonstrated by NMR (ROESY and PGSE) and mass spectrometry (ESI-TOF). These supramolecular complexes can be easily reduced to the corresponding more stable tetraamine macrocycles. Accordingly, we have used this reaction to efficiently synthesize a family of new pseudopeptidic macrocycles in a one-pot two-steps anion-templated reductive amination reaction, which comprises a multicomponent macrocyclization through the selective formation of four chemical bonds to yield a unique macrocyclic structure. Different variables like the aliphatic spacer between amino acidic moieties, geometry of the dialdehyde, and structure of the amino acid side chains were thoroughly studied, and their effect in the formation and stability of the supramolecular complexes discussed. The conformational preorganization induced by the template has been monitored by circular dichroism, reflecting the differences observed in the isolated yields, as well as by NMR spectroscopy. This effect has been also supported by molecular modeling. All the experimental and theoretical techniques were strongly consistent and reflected the same trends by comparing the different structural variables introduced in the system. PMID- 18402443 TI - A calcium-modulated plasmonic switch. AB - A plasmonic switch based on the calcium-induced conformational changes of calmodulin is shown to exhibit reversible wavelength modulations in response to changing calcium concentration. The extinction maximum (lambdamax) of a localized surface plasmon resonance (LSPR) sensor functionalized with a novel calmodulin construct, cutinase-calmodulin-cutinase (CutCaMCut), reversibly shifts by 2-3 nm. A high-resolution (HR) LSPR spectrometer with a wavelength resolution (3sigma) of 1.5 x 10-2 nm was developed to detect these wavelength modulations in real-time, providing information about the dynamics and structure of the protein. The rate of conversion from open (Ca2+-bound) to closed (Ca2+-free) calmodulin is shown to be 4-fold faster than the reverse process, with a closing rate of 0.127 s-1 and opening rate of 0.034 s-1. As far as we are aware, this plasmonic switch marks the first use of LSPR spectroscopy to detect reversible conformational changes in an unlabeled protein. PMID- 18402444 TI - Autocatalytic oxidative addition of PhBr to Pd(PtBu3)2 via Pd(PtBu3)2(H)(Br). AB - We report that oxidative addition of bromobenzene to Pd(PtBu3)2 occurs by an unusual autocatalytic mechanism. Studies on the effect of various additives showed that the degree of rate acceleration followed the trend: (PtBu3)Pd(Ph)(Br) approximately (HPtBu3)Br < [(PtBu3)Pd(mu-Br)]2 < (PtBu3)2Pd(H)(Br). Studies on the reactions of Pd(PtBu3)2 in the presence of (PtBu3)2Pd(H)(Br) showed that the concentration of (PtBu3)2Pd(H)(Br) decreased only after the Pd(0) complex had been consumed. These data indicated that the catalyst in this process is (PtBu3)2Pd(H)(Br). Thermal decomposition of the three-coordinate oxidative addition product (PtBu3)Pd(Ar)(Br) during the reaction of Pd(PtBu3)2 and bromoarenes ultimately leads to formation of (PtBu3)2Pd(H)(Br). Parallel reactions of bromobenzene with (PtBu3)2Pd(H)(Br) and Pd(PtBu3)2 showed that the bromoarenes reacted considerably faster with the Pd(II) species than with the Pd(0) species. We therefore propose a catalytic cycle for oxidative addition in which PBut3.HBr reacts with the Pd(0) species to form (PtBu3)2Pd(H)(Br), and (PtBu3)2Pd(H)(Br) reacts with the bromoarene, possibly though the anionic species [HPtBu3+][(PtBu3)Pd(Br)-], to form [Pd(PtBu3)(Ar)(Br)]. PMID- 18402445 TI - Functional determinants of a synthetic vesicle fusion system. AB - Selective membrane mergers may be driven by small-molecule recognition between synthetic surface-displayed fusogens which bear vancomycin glycopeptide and its native binding target, D-Ala-D-Ala dipeptide. These recognition motifs are membrane anchored by antimicrobial peptide magainin II and a phosphatidylethanolamine lipid derivative, respectively. We report herein characterization of this synthetic membrane fusion reaction with regard to the following: effects of fusogen concentration, lipid composition, and membrane charge. Our findings indicate that these parameters are determinants of fusion rate, vesicle stability, peptide binding, catalytic fusion and membrane disruption during fusion. Notably, these data indicate the importance of coupling between molecular recognition and insertion for bilayer activation as well as the critical role of membrane subdomain formation for membrane fusion reactivity. These phenomena are general to lipid membrane chemistry, and therefore these findings provide a guideline for understanding more complex biomembrane systems. PMID- 18402446 TI - Isolation and characterization of a neutral imino-semiquinone radical. AB - The neutral imino-semiquinone, 4,6-di-tert-butyl-2-tert-butylimino-semiquinone (isqH.), can be prepared by a conproportionation of the parent aminophenol and iminoquinone compounds. The neutral radical species has been characterized in the solid state by X-ray diffraction and in solution by EPR and UV-vis absorption spectroscopy. The stability of the open-shell radical is derived from the basicity of the tert-butylimino group and the intramolecular hydrogen bond. PMID- 18402447 TI - Deprotonation induced ligand-to-metal electron transfer: synthesis of a mixed valence Rh(-I,I) dinuclear compound and its reaction with dioxygen. AB - Treatment of bis(2-picolyl)amine (bpa) with [{Rh(nbd)(mu-OMe)}2] leads to the unexpected and unique redox asymmetric dinuclear Rh-I,Rh+I complex [{Rh(nbd)}2(bpa-2H)] (2) with a pi-coordinating imine bound to a tetrahedral low valent rhodate(-I). Mono-oxygenation of the deprotonated bpa ligand in 2 by O2 leads to the mononuclear carboxamido complex [Rh(nbd)(bpam-H)] (3) (bpam = N-(2 picolyl)picolinamide). The second O atom of O2 ends up as a hydroxo fragment in [{Rh(nbd)(mu-OH)}2]. PMID- 18402448 TI - Beta-H transfer from the metallacyclobutane: a key step in the deactivation and byproduct formation for the well-defined silica-supported rhenium alkylidene alkene metathesis catalyst. AB - The surface complex [([triple bond]SiO)Re([triple bond]CtBu)(=CHtBu)(CH2tBu)] (1) is a highly efficient propene metathesis catalyst with high initial activities and a good productivity. However, it undergoes a fast deactivation process with time on stream, which is first order in active sites and ethene. Noteworthy, 1 butene and pentenes, unexpected products in the metathesis of propene, are formed as primary products, in large amount relative to Re (>>1 equiv/Re), showing that their formation is not associated with the formation of inactive species. DFT calculations on molecular model systems show that byproduct formation and deactivation start by a beta-H transfer trans to the weak sigma-donor ligand (siloxy) at the metallacyclobutane intermediate having a square-based pyramid geometry. This key step has an energy barrier slightly higher than that calculated for olefin metathesis. After beta-H transfer, the most accessible pathway is the insertion of ethene in the Re-H bond. The resulting pentacoordinated trisperhydrocarbyl complex rearranges via either (1) alpha-H abstraction yielding the unexpected 1-butene byproduct and the regeneration of the catalyst or (2) beta-H abstraction leading to degrafting. These deactivation and byproduct formation pathways are in full agreement with the experimental data. PMID- 18402450 TI - Dynamics of silica-supported catalysts determined by combining solid-state NMR spectroscopy and DFT calculations. AB - The molecular dynamics of a series of organometallic complexes covalently bound to amorphous silica surfaces is determined experimentally using solid-state nuclear magnetic resonance (NMR) spectroscopy and density functional theory calculations (DFT). The determination is carried out for a series of alkylidene based catalysts having the general formula [([triple bond]SiO)M(ER)(=CH(t)Bu)(R')] (M = Re, Ta, Mo or W; ER = C(t)Bu, NAr or CH2(t)Bu; R' = CH2(t)Bu, NPh2, NC4H4). Proton-carbon dipolar coupling constants and carbon chemical shift anisotropies (CSA) are determined experimentally by solid-state NMR. Room-temperature molecular dynamics is quantified through order parameters determined from the experimental data. For the chemical shift anisotropy data, we validate and use a method that integrates static values for the CSA obtained computationally by DFT, obviating the need for low-temperature measurements. Comparison of the room-temperature data with the calculations shows that the widths of the calculated static limit dipolar couplings and CSAs are always greater than the experimentally determined values, providing a clear indication of motional averaging on the NMR time scale. Moreover, the dynamics are found to be significantly different within the series of molecular complexes, with order parameters ranging from = 0.5 for [([triple bond]SiO)Ta(=CH(t)Bu)(CH2(t)Bu)2] and [([triple bond]SiO)Re([triple bond]C(t)Bu)(=CH(t)Bu)(CH2(t)Bu)] to = 0.9 for [([triple bond]SiO)Mo([triple bond]NAr)(=CH(t)Bu)(R') with R' = CH2(t)Bu, NPh2, NC4H4. The data also show that the motion is not isotropic and could be either a jump between two sites or more likely restricted librational motion. The dynamics are discussed in terms of the molecular structure of the surface organometallic complexes, and the orientation of the CSAs tensor at the alkylidene carbon is shown to be directly related to the magnitude of the alpha-alkylidene CH agostic interation. PMID- 18402451 TI - Vapor-liquid-solid and vapor-solid growth of phase-change Sb2Te3 nanowires and Sb2Te3/GeTe nanowire heterostructures. AB - We report the synthesis and characterization of radial heterostructures composed of an antimony telluride (Sb2Te3) core and a germanium telluride (GeTe) shell, as well as an improved synthesis of Sb2Te3 nanowires. The synthesis of the heterostructures employs Au-catalyst-assisted vapor-liquid-solid (VLS) and vapor solid (VS) mechanisms. Energy-dispersive X-ray spectrometry indicates that Sb and Ge are localized in the Sb2Te3 and GeTe portions, respectively, confirming the alloy-free composition in the core/shell heterostructures. Transmission electron microscopy and diffraction studies show that Sb2Te3 and GeTe regions exhibit rhombohedral crystal structure. Both Sb2Te3 and GeTe grow along the [110] direction with an epitaxial interface between them. Electrical characterization of individual nanowires and nanowire heterostructures demonstrates that these nanostructures exhibit memory-switching behavior. PMID- 18402449 TI - Noncovalent cell surface engineering: incorporation of bioactive synthetic glycopolymers into cellular membranes. AB - The controlled addition of structurally defined components to live cell membranes can facilitate the molecular level analysis of cell surface phenomena. Here we demonstrate that cell surfaces can be engineered to display synthetic bioactive polymers at defined densities by exogenous membrane insertion. The polymers were designed to mimic native cell-surface mucin glycoproteins, which are defined by their dense glycosylation patterns and rod-like structures. End-functionalization with a hydrophobic anchor permitted incorporation into the membranes of live cultured cells. We probed the dynamic behavior of cell-bound glycopolymers bearing various hydrophobic anchors and glycan structures using fluorescence correlation spectroscopy (FCS). Their diffusion properties mirrored those of many natural membrane-associated biomolecules. Furthermore, the membrane-bound glycopolymers were internalized into early endosomes similarly to endogenous membrane components and were capable of specific interactions with protein receptors. This system provides a platform to study cell-surface phenomena with a degree of chemical control that cannot be achieved using conventional biological tools. PMID- 18402452 TI - Ruthenium nanoparticles inside porous [Zn4O(bdc)3] by hydrogenolysis of adsorbed [Ru(cod)(cot)]: a solid-state reference system for surfactant-stabilized ruthenium colloids. AB - The gas-phase loading of [Zn4O(bdc)3] (MOF-5; bdc = 1,4-benzenedicarboxylate) with the volatile compound [Ru(cod)(cot)] (cod = 1,5-cyclooctadiene, cot = 1,3,5 cyclooctatriene) was followed by solid-state (13)C magic angle spinning (MAS) NMR spectroscopy. Subsequent hydrogenolysis of the adsorbed complex inside the porous structure of MOF-5 at 3 bar and 150 degrees C was performed, yielding ruthenium nanoparticles in a typical size range of 1.5-1.7 nm, embedded in the intact MOF-5 matrix, as confirmed by transmission electron microscopy (TEM), selected area electron diffraction (SAED), powder X-ray diffraction (PXRD), and X-ray absorption spectroscopy (XAS). The adsorption of CO molecules on the obtained Ru@MOF-5 nanocomposite was followed by IR spectroscopy. Solid-state (2)H NMR measurements indicated that MOF-5 was a stabilizing support with only weak interactions with the embedded particles, as deduced from the surprisingly high mobility of the surface Ru-D species in comparison to surfactant-stabilized colloidal Ru nanoparticles of similar sizes. Surprisingly, hydrogenolysis of the [Ru(cod)(cot)]3.5@MOF-5 inclusion compound at the milder condition of 25 degrees C does not lead to the quantitative formation of Ru nanoparticles. Instead, formation of a ruthenium-cyclooctadiene complex with the arene moiety of the bdc linkers of the framework takes place, as revealed by (13)C MAS NMR, PXRD, and TEM. PMID- 18402453 TI - Ribosomal synthesis of N-methyl peptides. AB - N-methyl amino acids (N-Me AAs) are a common component of nonribosomal peptides (NRPs), a class of natural products from which many clinically important therapeutics are obtained. N-Me AAs confer peptides with increased conformational rigidity, membrane permeability, and protease resistance. Hence, these analogues are highly desirable building blocks in the ribosomal synthesis of unnatural peptide libraries, from which functional, NRP-like molecules may be identified. By supplementing a reconstituted Escherichia coli translation system with specifically aminoacylated total tRNA that has been chemically methylated, we have identified three N-Me AAs (N-Me Leu, N-Me Thr, and N-Me Val) that are efficiently incorporated into peptides by the ribosome. Moreover, we have demonstrated the synthesis of peptides containing up to three N-Me AAs, a number comparable to that found in many NRP drugs. With improved incorporation efficiency and translational fidelity, it may be possible to synthesize combinatorial libraries of peptides that contain multiple N-Me AAs. Such libraries could be subjected to in vitro selection methods to identify drug-like, high-affinity ligands for protein targets of interest. PMID- 18402454 TI - Leaching and efficiency of six organic zinc fertilizers applied to navy bean crop grown in a weakly acidic soil of Spain. AB - Zinc contamination of groundwater from fertilizers applied to pulse crops is a potential problem, but the use of different types of organic chelates can minimize the contamination potential while still adequately feeding the crops. The objective of this study was to compare the leaching, distribution in fractions and availability, and relative effectiveness of Zn from six organic Zn fertilizers (zinc-ethylenediaminetetraacetate- N-2 hydroxyethylethylenediaminetriacetate (Zn-EDTA-HEDTA), Zn-HEDTA, zinc- S, S' ethylenediaminedisuccinate (Zn- S, S-EDDS), zinc-polyhydroxyphenylcarboxylate, Zn EDTA, and zinc-ethylenediaminedi(2-hydroxy-5-sulfophenylacetate) (Zn-EDDHSA)) applied to a navy bean ( Phaseolus vulgaris, L.) crop cultivated by applying different Zn levels, in a weakly acidic soil under greenhouse conditions. Zinc soil behavior was evaluated by diethylenetriaminepentaacetic acid-triethanolamine (DTPA-TEA), DTPA-ammonium bicarbonate (DTPA-AB), Mehlich-3, and BaCl 2 extractions and sequential fractionation. In all the fertilizer treatments, the percentage of labile Zn that remained in the soil was high with respect to the quantity of Zn applied, with values respectively ranging from 42 to 80% for Zn EDDHSA and Zn-EDTA sources. A positive correlation with a high level of significance existed between the micronutrient concentration in the navy bean crop (total and soluble) and labile Zn fractions, available Zn, and easily leachable Zn ( r ranged from 0.89 to 0.95, P < 0.0001). The relatively high quantity of total Zn leached by applying Zn-EDTA and Zn-S,S-EDDS sources (11.9 and 6.0%, respectively, for the rate 10 mg of Zn kg(-1) of soil) poses a potential pollution risk for neighboring waters. It would seem recommendable to apply Zn-HEDTA or Zn-EDDHSA sources, even applied at the low rate (5 mg of Zn kg( 1) of soil), because they produced available Zn concentrations in the soil that were above the critical concentration and also produced high Zn concentrations in plants (139 and 106 mg of Zn kg(-1) of dry matter, respectively). PMID- 18402455 TI - Total synthesis of (-)-kainic acid via intramolecular Michael addition: a second generation route. AB - A total synthesis of (-)-kainic acid starting from the commercially available 2 azetidinone is described. The key delta-lactone intermediate was concisely prepared from the commercially available azetidinone through the Reformatsky-type reaction and an introduction of a glycine moiety. The construction of the functionalized pyrrolidine ring was executed by a one-pot sequential elimination Michael addition protocol of a beta-amino-delta-lactone intermediate with high diastereoselectivity. PMID- 18402456 TI - Rhodium-catalyzed borylative cyclization of 2-alkynylaryl isocyanates with bis(pinacolato)diboron. AB - 2-alkynylaryl isocyanates reacted with bis(pinacolato)diboron in the presence of a cationic rhodium(I) catalyst to produce borylated 3-alkylideneoxindoles stereoselectively. The carbon-boron linkage formed was further transformed via reactions such as a cross-coupling and a halogenation reaction. PMID- 18402457 TI - A cation-directed switch of intermolecular spin-spin interaction of guanosine derivatives functionalized with open-shell units. AB - The guanosine derivative 1 functionalized with the persistent radical unit 4 carbonyl-2,2,6,6-tetramethylpiperidin-1-oxyl in solution has no particular intermolecular spin-spin interactions; however, in the presence of potassium ions this compound can form a D4-symmetric octameric assembly [1(8)K]+ in which the nitroxyl moieties show a weak electron spin-spin exchange interaction. Since the relative geometry of the radicals is the outcome of K+-directed self-assembly, the spin-spin interaction can be suppressed by removing the alkaline ion. PMID- 18402458 TI - Palladium-catalyzed carbene insertion and trapping with carbon nucleophiles. AB - Palladium catalysts are shown to catalyze the three-component coupling of vinyl halides, trimethylsilyldiazomethane, and stabilized carbon nucleophiles. The reaction is believed to proceed through a palladium-carbene intermediate LX(R)PdCHSiMe 3 that undergoes migration of the vinyl substituent to the electrophilic carbene center to generate an eta 3-allylpalladium intermediate. The allylpalladium intermediate is attacked by the carbon nucleophile to generate a vinylsilane product. PMID- 18402459 TI - Gas-phase stereoselective binding to Mn/salen catalysts. AB - Gas-phase equilibrium measurements have been used to determine the stereoselectivity of binding the enantiomers of 1-phenylethanol to manganese/salen asymmetric epoxidation catalysts. There is significant selectivity in the gas-phase binding, and the results are compared to data from condensed-phase epoxidations. The study demonstrates the utility of a novel internal standard approach that allows for rapid, accurate measures of the stereoselectivity of gas-phase ligand binding. Moreover, the data suggest that gas-phase binding stereoselectivity could be a potential predictor of condensed phase enantioselectivity. PMID- 18402460 TI - Enantiopure 1,5-diols from dynamic kinetic asymmetric transformation. Useful synthetic intermediates for the preparation of chiral heterocycles. AB - Dynamic kinetic asymmetric transformation (DYKAT) of a series of 1,5-diols has been performed in the presence of Candida antarctica lipase B (CALB), Pseudomonas cepacia lipase II (PS-C II), and ruthenium catalyst 4. The resulting optically pure 1,5-diacetates are useful synthetic intermediates, which was demonstrated by the syntheses of both an enantiopure 2,6-disubstituted piperidine and an enantiopure 3,5-disubstituted morpholine. PMID- 18402461 TI - Synthesis and stereochemistry-activity relationship of small bacteriocin, an autoinducer of the symbiotic nitrogen-fixing bacterium Rhizobium leguminosarum. AB - The four stereoisomers of small bacteriocin, an autoinducer of the symbiotic nitrogen-fixing bacterium Rhizobium leguminosarum, were synthesized via a versatile methodology for 3'-hydroxyacyl homoserine lactones based on the Nagao asymmetric aldol reaction. The synthetic isomers were much less effective at inhibiting the growth of R. leguminosarum RBL5523 than the natural isomer, showing the importance of stereochemistry for activity. PMID- 18402462 TI - Benzyl protection of phenols under neutral conditions: palladium-catalyzed benzylations of phenols. AB - Benzyl protection of phenols under neutral conditions was achieved by using a Pd(eta3-C3H5)Cp-DPEphos catalyst. The palladium catalyst efficiently converted aryl benzyl carbonates into benzyl-protected phenols through the decarboxylative etherification. Alternatively, the nucleophilic substitution of benzyl methyl carbonates with phenols proceeded in the presence of the catalyst, yielding aryl benzyl ethers. PMID- 18402463 TI - Total synthesis of (+/-)-murrayazoline. AB - The total synthesis of (+/-)-murrayazoline (1) is described. The characteristic hexa-heterocyclic structure of 1 was constructed by a combination of the intramolecular Friedel-Crafts-type Michael addition and Pd-catalyzed C-O coupling reactions. The N-substituted carbazole component was synthesized in one pot by the double N-arylation of a sterically hindered amine with a dibromobiphenyl derivative. PMID- 18402464 TI - Towards the rational synthesis of norfullerenes. controlled deletion of one carbon atom from C60 and preparation of 2,5,9-trioxo-1-nor(C60-Ih)[5,6]fullerene C59(O)3 derivatives. AB - Norfullerenes are fullerene-like compounds (fulleroids) resulting from partial deletion of fullerene skeleton carbons. The one carbon less norfullerene C59(O) 3 derivatives having three carbonyl groups on the rim of the orifice are prepared through peroxide-mediated reactions. A key step involves a novel PCl 5 initiated rearrangement of a hydroxyl amine adduct. Decarboxylation serves as the carbon removal step. PMID- 18402467 TI - Comparative NMR analysis of cellooligosaccharide hydrolysis by GH9 bacterial and plant endo-1,4-beta-glucanases. AB - 1H NMR spectroscopy has been used to analyze the product profiles arising from the hydrolysis of cellooligosaccharides by family GH9 cellulases. The product profiles obtained with the wild type and several active site mutants of a bacterial processive endoglucanase, TfCel9A, were compared with those obtained by a randomly acting plant endoglucanase, PttCel9A. PttCel9A is an orthologue of the Arabidopsis endocellulase, Korrigan, which is required for efficient cellulose biosynthesis. As expected, poplar PttCel9A was shown to catalyze the degradation of cellooligosaccharides by inversion of the configuration of the anomeric carbon. The product analyses showed that the number of interactions between the glucose units of the substrate and the aromatic residues in the enzyme active sites determines the point of cleavage in both enzymes. PMID- 18402465 TI - Purification and reconstitution of sterol transfer by native mouse ABCG5 and ABCG8. AB - ABCG5 (G5) and ABCG8 (G8) are ATP-binding cassette half-transporters that limit intestinal uptake and promote biliary secretion of neutral sterols. Here, we describe the purification of endogenous G5G8 from mouse liver to near homogeneity. We incorporated the native proteins into membrane vesicles and reconstituted sterol transfer. Native gel electrophoresis, density-gradient ultracentrifugation, and chemical cross-linking studies indicated that the functional native complex is a heterodimer. No higher order oligomeric forms were observed at any stage in the catalytic cycle. Sterol transfer activity by purified native G5G8 was stable, stereospecific, and selective. We also report that G5 but not G8 is S-palmitoylated and that palmitoylation is not essential for dimerization, trafficking, or biliary sterol secretion. Both G5 and G8 have short but highly conserved cytoplasmic tails. The functional roles of these C terminal regions were examined using an in vivo functional assay. PMID- 18402468 TI - Xenobiotic geometry and media pH determine cytotoxicity through solubility. AB - Polychlorinated biphenyls (PCBs), a class of 209 individual congeners, have become persistent and ubiquitous environmental contaminants. The health impacts of PCBs, such as cancer, cardiovascular disease, developmental toxicity, and neurotoxicity, have been widely reported, but for many of these, the mechanisms of toxicity are still poorly understood. Many mechanistic studies involve cultured cells where the biological activity is dependent upon the solubility of the xenobiotic. In the present study, we investigated the factors that determine solubility as measured by diffraction spectroscopy and have modeled, with semiempirical and ab initio molecular orbital methods, the dihedral angle and calculated the dipole moment of a series of monofluorinated analogues (F-PCBs 3) of 4-chlorobiphenyl (PCB 3) as model compounds in vacuum and in water. We found a strong positive correlation between the dihedral angle, the rotation energy, the cavitation energy, the solubility, and the cytotoxicity in three human cell lines. The dipole moment was of minor influence. We also determined the influence of pH changes in a medium containing 10% fetal bovine serum (FBS), changes that could be expected when cells in culture are removed from a CO 2 incubator even for a short time. We found that the solubility is strongly affected by the pH and that this effect is not reversed by subsequent pH readjustment. In a study examining cytotoxicity, we showed that the actual pH and the pH history of a medium containing FBS were of major influence. We suggest that pH-driven changes in the tertiary and quaternary structure of albumin are responsible. These observations have implications for studies of the biological activity of semisoluble compounds, like PCBs and related compounds. PMID- 18402469 TI - Comparisons of (+/-)-benzo[a]pyrene-trans-7,8-dihydrodiol activation by human cytochrome P450 and aldo-keto reductase enzymes: effect of redox state and expression levels. AB - Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants that are metabolically activated to proximate carcinogenic trans-dihydrodiols. PAH trans dihydrodiols are further activated in humans by cytochrome P450 (P450) 1A1 and 1B1 to yield diol-epoxides or by aldo-keto reductases (AKR) 1A1 and 1C1-1C4 to yield reactive and redox-active o-quinones. Reconstituted in vitro systems were used to compare the steady-state kinetic constants for human P450 (P450 1A1 and 1B1) and AKR (AKR1A1, AKR1C1-1C4) mediated metabolism of (+/-)- trans-7,8 dihydroxy-7,8-dihydrobenzo[ a]pyrene ((+/-)-B[ a]P-7,8-diol) at physiological pH. It was found that P450 isoforms yielded much greater k cat/ K m values than AKR enzymes. Initial rates of (+/-)-B[ a]P-7,8-diol oxidation were measured for AKR1A1, AKR1C2, P450 1A1, and P450 1B1 as the ratio of NADPH/NAD (+) cofactors was varied to determine the redox state necessary for AKRs to successfully compete for trans-dihydrodiols. P450 and AKR enzymes equally competed for (+/-) B[ a]P-7,8-diol substrate at an NADPH/NAD (+) ratio equal to 0.001. The resting NADPH/NAD (+) ratio was determined in A549 human lung adenocarcinoma cells to be 0.28. These data suggest that the P450 pathway would be favored over the AKR pathway if the enzymes were equally expressed. Basal mRNA transcript levels of AKR1C1-1C3 exceed those of both basal and 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD)-induced P450 1A1 and 1B1 by up to 90-fold in A549 cells as measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR) methods. AKR expression levels were comparable to TCDD-induced P450 1A1 and 1B1 in HBEC-KT immortalized normal human bronchial epithelial cells. Functional assays of both A549 and HBEC-KT cell lysates demonstrated a lack of TCDD-inducible P450 1A1/1B1 activity but robust basal expression of AKR1A1 and AKR1C activities, where the functional assay for P450 detection is 300-fold more sensitive than the functional assay for AKR isoforms. These data suggest that AKR enzymes may effectively compete with P450 1A1/1B1 for PAH trans-dihydrodiol activation in human lung cells. PMID- 18402471 TI - All-conjugated block copolymers. AB - All-conjugated block copolymers of the rod-rod type came into the focus of interest because of their unique and attractive combination of nanostructure formation and electronic activity. Potential applications in a next generation of organic polymer materials for photovoltaic devices ("bulk heterojunction"-type solar cells) or (bio)sensors have been proposed. Combining the fascinating self assembly properties of block copolymers with the active electronic and/or optical function of conjugated polymers in all-conjugated block copolymers is, therefore, a very challenging goal of synthetic polymer chemistry. First examples of such all-conjugated block copolymers from a couple of research groups all over the world demonstrate possible synthetic approaches and the rich application potential in electronic devices. A crucial point in such a development of novel polymer materials is a rational control over their nanostructure formation. All conjugated di- or triblock copolymers may allow for an organization of the copolymer materials into large-area ordered arrays with a length scale of nanostructure formation of the order of the exciton diffusion length of organic semiconductors (typically ca. 10 nm). Especially for amphiphilic, all-conjugated copolymers the formation of well-defined supramolecular structures (vesicles) has been observed. However, intense further research is necessary toward tailor-made, all-conjugated block copolymers for specific applications. The search for optimized block copolymer materials should consider the electronic as well as the morphological (self-assembly) properties. PMID- 18402470 TI - Bifunctional asymmetric catalysis: cooperative Lewis acid/base systems. AB - In the field of catalytic, asymmetric synthesis, there is a growing emphasis on multifunctional systems, in which multiple parts of a catalyst or multiple catalysts work together to promote a specific reaction. These efforts, in part, are result-driven, and they are also part of a movement toward emulating the efficiency and selectivity of nature's catalysts, enzymes. In this Account, we illustrate the importance of bifunctional catalytic methods, focusing on the cooperative action of Lewis acidic and Lewis basic catalysts by the simultaneous activation of both electrophilic and nucleophilic reaction partners. For our part, we have contributed three separate bifunctional methods that combine achiral Lewis acids with chiral cinchona alkaloid nucleophiles, for example, benzoylquinine (BQ), to catalyze highly enantioselective cycloaddition reactions between ketene enolates and various electrophiles. Each method requires a distinct Lewis acid to coordinate and activate the electrophile, which in turn increases the reaction rates and yields, without any detectable influence on the outstanding enantioselectivities inherent to these reactions. To place our results in perspective, many important contributions to this emerging field are highlighted and our own reports are chronicled. PMID- 18402472 TI - Nanomechanical fingerprints of individual blocks of a diblock copolymer chain. AB - Atomic force microscopy (AFM)-based single polymer chain pulling experiments have been used to study the structural transitions of individual homopolymer chains in water. Polystyrene (PS) showed a three-regime force-extension profile exhibiting a force plateau reminiscent of a first-order transition, as predicted theoretically, whereas poly(methyl methacrylate) (PMMA) showed a characteristic saw-tooth pattern reminiscent of multidomain disassembly behavior. The two distinct structural transtions provide fingerprints for the individual homopolymers, which can be used to identify individual blocks of symmetric and asymmetric PS- b-PMMA diblock copolymer chains. PMID- 18402473 TI - Electrochemical characterization of pore formation by bacterial protein toxins on hybrid supported membranes. AB - The interaction of pore-forming streptolysin O (SLO) with biomimetic lipid membranes has been studied by electrochemical methods. Phosphatidylcholine lipid vesicles were deposited onto gold electrodes modified with supporting layers of hexyl thioctate (HT) or thioctic acid tri(ethylene glycol) ester (TA-TEGE), and integrity and permeability of the resulting membranes were characterized by cyclic voltammetry and impedance spectroscopy. Both positively and negatively charged electrochemical probes, potassium ferrocyanide, hexaammineruthenium(III) chloride, and ferrocene carboxylic acid (FCA), were employed to evaluate their suitability to probe the membrane permeability properties, with FCA exhibiting ideal behavior and thus employed throughout the work. Fusion of vesicles incubated with SLO on the electrodes yielded membranes that showed a distinctive response pattern for FCA as a function of SLO concentration. A direct dependence of both the currents and peak separation of FCA in the cyclic voltammograms was observed over a concentration range of 0-10 hemolytic units (HU)/microL of the toxin. The interaction of SLO with preformed supported lipid membranes was also investigated, and much lower response was observed, suggesting a different extent of membrane-toxin interactions on such an interface. Nonionic surfactant Triton was found to disrupt the vesicle structure but could not completely remove a preformed membrane to fully restore the electrode response. The information reported here offers some unique insight into toxin-surface interactions on a hybrid membrane, facilitating the development of electrochemically based sensing platforms for detecting trace amounts of bacterial toxins via the perforation process. PMID- 18402475 TI - pH-dependent locking of giant mesogens in fibers drawn from mussel byssal collagens. AB - Byssal threads are tough collagenous fibers that mussels use to secure themselves against dislodgement by waves in the marine intertidal zone. Here, preCol, a family of hybrid collagens comprising up to 96% of the protein content in certain regions of byssal threads, was purified in mg amounts from mussel foot tissue for the first time. Conditions for drawing preCols into quality fibers ex vivo were investigated. The most important factor affecting fiber formation was the pH of the drawing solution. The morphology and tensile properties of drawn fibers were also characterized and suggest that a liquid crystal mesophase combined with cross-linking by His-metal coordination plays a role in the assembly/mechanics of drawn fibers and likely in native byssal threads as well. PMID- 18402476 TI - An expedient synthesis of mellitic triimides. AB - Heating of the solid ammonium salts obtained from treatment of mellitic acid with 3 equiv of a primary amine yields trisubstituted mellitic triimides via dehydration and imide ring closure. This surprisingly simple synthetic approach is amenable to incorporation of alkyl, aryl, and amino acid ester substituents, thereby opening broad access to a family of C(3)-symmetric organic electron acceptors. PMID- 18402477 TI - Strain-driven electronic band structure modulation of si nanowires. AB - One of the major challenges toward Si nanowire (SiNW) based photonic devices is controlling the electronic band structure of the Si nanowire to obtain a direct band gap. Here, we present a new strategy for controlling the electronic band structure of Si nanowires. Our method is attributed to the band structure modulation driven by uniaxial strain. We show that the band structure modulation with lattice strain is strongly dependent on the crystal orientation and diameter of SiNWs. In the case of [100] and [111] SiNWs, tensile strain enhances the direct band gap characteristic, whereas compressive strain attenuates it. [110] SiNWs have a different strain dependence in that both compressive and tensile strain make SiNWs exhibit an indirect band gap. We discuss the origin of this strain dependence based on the band features of bulk silicon and the wave functions of SiNWs. These results could be helpful for band structure engineering and analysis of SiNWs in nanoscale devices. PMID- 18402478 TI - Imaging mechanical vibrations in suspended graphene sheets. AB - We carried out measurements on nanoelectromechanical systems based on multilayer graphene sheets suspended over trenches in silicon oxide. The motion of the suspended sheets was electrostatically driven at resonance using applied radio frequency voltages. The mechanical vibrations were detected using a novel form of scanning probe microscopy, which allowed identification and spatial imaging of the shape of the mechanical eigenmodes. In as many as half the resonators measured, we observed a new class of exotic nanoscale vibration eigenmodes not predicted by the elastic beam theory, where the amplitude of vibration is maximum at the free edges. By modeling the suspended sheets with the finite element method, these edge eigenmodes are shown to be the result of nonuniform stress with remarkably large magnitudes (up to 1.5 GPa). This nonuniform stress, which arises from the way graphene is prepared by pressing or rubbing bulk graphite against another surface, should be taken into account in future studies on electronic and mechanical properties of graphene. PMID- 18402479 TI - Establishing effective simulation protocols for beta- and alpha/beta-peptides. II. Molecular mechanical (MM) model for a cyclic beta-residue. AB - All-atom molecular mechanical (MM) force field parameters are developed for a cyclic beta-amino acid, amino-cyclo-pentane-carboxylic acid (ACPC), using a multi objective evolutionary algorithm. The MM model is benchmarked using several short, ACPC-containing alpha/beta-peptides in water and methanol with SCC-DFTB (self consistent charge-density functional tight binding)/MM simulations as the reference. Satisfactory agreements are found between the MM and SCC-DFTB/MM results regarding the distribution of key dihedral angles for the tetra alpha/beta-peptide in water. For the octa-alpha/beta-peptide in methanol, the MM and SCC-DFTB/MM simulations predict the 11- and 14/15-helical form as the more stable conformation, respectively; however, the two helical forms are very close in energy (2-4 kcal/mol) at both theoretical levels, which is also the conclusion from recent NMR experiments. As the first application, the MM model is applied to an alpha/beta-pentadeca-peptide in water with both explicit and implicit solvent models. The stability of the peptide is sensitive to the starting configuration in the explicit solvent simulations due to their limited length ( approximately 10-40 ns). Multiple ( approximately 20 x 20 ns) implicit solvent simulations consistently show that the 14/15-helix is the predominant conformation of this peptide, although substantially different conformations are also accessible. The calculated nuclear Overhauser effect (NOE) values averaged over different trajectories are consistent with experimental data, which emphasizes the importance of considering conformational heterogeneity in such comparisons for highly dynamical peptides. PMID- 18402480 TI - Methimazole-based ionic liquids. AB - The alkylation reaction of 2-mercapto-1-methylimidazole 1a with iodoethane and chlorobutane produced S-alkylmethimazole halides 2a and 2b which were subjected to anion metathesis with two different metal salts (MA) to afford methimazole based room-temperature ionic liquids 3a, 3b, and 3c in 82%, 85%, and 87% yields, respectively. S-Alkylation giving 2a and 2b suggests that methimazole reacts through the thione tautomer. PMID- 18402481 TI - Is a proposed reaction mechanism free from unnecessary assumptions? Occam's razor applied in a mathematical way to complex first-order reaction systems. AB - Following Occam's principle, a proposed reaction mechanism should not contain assumptions about the existence of reactive intermediates and reaction paths that are unnecessary for a full description and interpretation of the available facts. A mechanism refers, in this paper, to a proposed reaction scheme or network that represents the reactions supposed to be going on in a complex reaction system with observable species as well as unobservable reactive intermediates. The scope is limited here to (pseudo) first-order reactions and the steady-state approximation is invoked in order to relate unknown mechanistic rate constants to experimentally determined ones, and, when available, theoretically calculated quantities. When the resulting, nonlinear system of equations admits a unique solution within a physically reasonable domain, it is concluded that the reaction mechanism fulfills Occam's principle. Otherwise, there are many or no solutions. No subjective or qualitative arguments enter the procedure and the outcome is not negotiable. PMID- 18402482 TI - Evidence for emergent chemical bonding in Au+-Rg complexes (Rg = Ne, Ar, Kr, and Xe). AB - Evidence is presented that there is a clear covalent component in the bonding of Au+ to Kr and Au+ to Xe, with some evidence that there may be such bonding between Au+ and Ar; for Au+ and Ne, there is no such evidence, and the bonding seems to be entirely physical. A model potential analysis shows that when all attractive inductive and dispersive terms out to R-8 are properly included in the Au+-Ne case, with an Ae(-bR) Born-Mayer repulsive term, essentially all the bonding in Au+-Ne can be rationalized by physical attraction alone. This is consistent with a natural bond order (NBO) analysis of the Au+-Ne ab initio wavefunctions, which shows the charge on Au+ to be very close to 1.0. In contrast, similar model potential and NBO analyses show quite clearly that physical interactions alone cannot account for the large bond energy values for the Au+-Kr and Au+-Xe complexes and are consistent with covalent contributions to the Au+-Kr and Au+-Xe interactions. Au+-Ar is seen to lie on the borderline between these two limits. In performing the model potential analyses, high-level ab initio calculations are employed [CCSD(T) energies, extrapolated to the complete basis set limit], to obtain reliable values of Re, De and omegae as input. A comparison of the gold-Xe bond distances in several solid-state Au(I, II and III) oxidation-state complex ions, containing "ligand" Xe atoms, prepared by Seppelt and co-workers, with that of the "free" Au+-Xe gas-phase ion is made, and a discussion of the trends is presented. PMID- 18402483 TI - Entropic changes control the charge separation process in triads mimicking photosynthetic charge separation. AB - Laser-induced optoacoustic spectroscopy (LIOAS) measurements with carotene porphyrin-acceptor "supermolecular" triads (C-P-A, with A = C60, a naphthoquinone NQ, and a naphthoquinone derivative, Q) were carried out with the purpose of analyzing the thermodynamic parameters for the formation and decay of the respective long-lived charge separated state C*+-P-A*-. The novel procedure of inclusion of the benzonitrile solutions of the triads in Triton X-100 micelle nanoreactors suspended in water permitted the separation of the enthalpic and structural volume change contributions to the LIOAS signals, by performing the measurements in the range 4-20 degrees C. Contractions of 4.2, 5.7, and 4.2 mL mol-1 are concomitant with the formation of C*+-P-A*- for A = C60, Q and NQ, respectively. These contractions are mostly attributed to solvent movements and possible conformational changes upon photoinduced electron transfer, due to the attraction of the oppositely charged ends, as a consequence of the giant dipole moment developed in these compounds upon charge separation ( approximately 110 D). The estimations combining the calculated free energies and the LIOAS-derived enthalpy changes indicate that entropy changes, attributed to solvent movements, control the process of electron transfer for the three triads, especially for C-P C60 and C-P-Q. The heat released during the decay of 1 mol of charge separated state (CS) is much smaller than the respective enthalpy content obtained from the LIOAS measurements for the CS formation. This is attributed to the production of long-lived energy storing species upon CS decay. PMID- 18402484 TI - Experimental and theoretical analysis of the reorientational dynamics of fullerene C70 in various aromatic solvents. AB - A previous study of C70 in deuterated chlorobenzene generated evidence suggesting C70 was experiencing unique reorientational behavior at given temperatures. The present study explores the possibility that this behavior is present across other solvents. The 13C spin-lattice relaxation rates for four carbon resonances in C70 were analyzed in benzene-d6, chlorobenzene-d5, and o-dichlorobenzene-d4, and as a function of temperature, to probe the reorientational dynamics of this fullerene. Anisotropic behavior was observed at the lowest (283 K) and highest temperatures (323 K), isotropic diffusion was seen between 293 and 303 K, and quasi-isotropic at 313 K. When anisotropic motion was present, diffusion about the figure axis was seen to be higher than diffusion of the figure axis. Experimentally obtained diffusion coefficients generated reorientational correlation times that were in excellent agreement with experimental values. Theoretical predictions generated by a modified Gierer-Wirtz model provided acceptable predictions of the diffusion constants; with DX usually being more closely reproduced and DZ values generally being underestimated. Overall, the results indicate that the factors affecting rotational behavior are complex and that multiple solvent factors are necessary to characterize the overall motion of C70 in these solvents. Although a solvent's viscosity is normally sufficient to characterize the tumbling motion, the spinning motion is less sensitive to solvent viscosity but more responsive to solvent structure. The balance and collective influence of these factors ultimately determines the overall rotational behavior. PMID- 18402485 TI - Physicochemical properties and microstructure formation of the surfactant mixtures of sodium N-(2-(n-dodecylamino)ethanoyl)-L-alaninate and SDS in aqueous solutions. AB - Aggregation behavior of a novel anionic amphiphilic molecule, sodium N-(2-(n dodecylamino)ethanoyl)-L-alaninate (C(12)Ala), was studied in the presence of sodium dodecyl sulfate (SDS) surfactant at different [C(12)Ala]/[SDS] molar ratios and concentrations. The viscosity of aqueous SDS solution increased in the presence of C(12)Ala surfactant. The bulk viscosity of water was found to increase upon increase of both molar ratio and total surfactant concentration. The microenvironments of the self-assemblies were investigated using the fluorescence probe technique. Fluorescence anisotropy studies indicated formation of rodlike micelles. Both dynamic light scattering and small-angle neutron scattering measurements were performed to obtain the size and shape of the microstructures. The concentration and composition dependence of the hydrodynamic diameter of the aggregates were investigated. Transmission electron micrographs revealed the presence of a hexagonal liquid crystal phase in dilute solutions of the C(12)Ala-SDS mixture. The micrographs of moderately concentrated solution, however, showed cholesteric liquid-crystal structures with fingerprint-like texture. Temperature-dependent phase behavior of the self-assemblies was studied by use of the fluorescence probe technique. PMID- 18402486 TI - Preparation of N,N'-bisethoxyethane[12]amideferrocenophane and its application in anion recognition. AB - Novel N,N'-bisethoxyethane[12]amideferrocenophane has been synthesized by a condensation reaction and characterized by (1)H NMR and mass spectrum. The anion recognition properties of the compound are evaluated via (1)H NMR, FT-IR, and electrochemical measurement. It is found that N,N' bisethoxyethane[12]amideferrocenophane exhibits remarkable electrochemical response to H(2)PO(4-) anion in CH(2)C(l2) or CH(3)CN solution, and response to anions can also be observed in CH(3)CN solution containing up to 15% water. Binding constants between the compound and HSO(4-) in different solutions have been determined by UV-vis spectrum titration experiments. The results indicate that the compound shows a selective recognition trend of H(2)PO(4-) > HSO(4-) (F( )) > Br(-). PMID- 18402487 TI - Temperature dependence of electron transfer to the M-side bacteriopheophytin in rhodobacter capsulatus reaction centers. AB - Subpicosecond time-resolved absorption measurements at 77 K on two reaction center (RC) mutants of Rhodobacter capsulatus are reported. In the D(LL) mutant the D helix of the M subunit has been substituted with the D helix from the L subunit, and in the D(LL)-FY(L)F(M) mutant, three additional mutations are incorporated that facilitate electron transfer to the M side of the RC. In both cases the helix swap has been shown to yield isolated RCs that are devoid of the native bacteriopheophytin electron carrier HL (Chuang, J. I.; Boxer, S. G.; Holten, D.; Kirmaier, C. Biochemistry 2006, 45, 3845-3851). For D(LL), depending whether the detergent Deriphat 160-C or N-lauryl-N,N-dimethylamine-N-oxide (LDAO) is used to suspend the RCs, the excited state of the primary electron donor (P*) decays to the ground state with an average lifetime at 77 K of 330 or 170 ps, respectively; however, in both cases the time constant obtained from single exponential fits varies markedly as a function of the probe wavelength. These findings on the D(LL) RC are most easily explained in terms of a heterogeneous population of RCs. Similarly, the complex results for D(LL)-FY(L)F(M) in Deriphat glycerol glass at 77 K are most simply explained using a model that involves (minimally) two distinct populations of RCs with very different photochemistry. Within this framework, in 50% of the D(LL)-FY(L)F(M) RCs in Deriphat-glycerol glass at 77 K, P* deactivates to the ground state with a time constant of approximately 400 ps, similar to the deactivation of P* in the D(LL) mutant at 77 K. In the other 50% of D(LL)-FY(L)F(M) RCs, P* has a 35 ps lifetime and decays via electron transfer to the M branch, giving P+HM- in high yield (> or =80%). This result indicates that P* --> P(+)H(M)(-) is roughly a factor of 2 faster at 77 K than at 295 K. In alternative homogeneous models the rate of this M-side electron-transfer process is the same or up to 2-fold slower at low temperature. A 2-fold increase in rate with a reduction in temperature is the same behavior found for the overall L-side process P* --> P(+)H(L)(-) in wild-type RCs. Our results suggest that, as for electron transfer on the L side, the M-side electron transfer reaction P* --> P(+)H(M)(-) is an activationless process. PMID- 18402488 TI - Competing patterns of weak directional forces in pressure-frozen CH2ClI and CH2I2. AB - Isostructural relations and phase transitions of dihalomethanes have been rationalized by the competing patterns of CH...halogen hydrogen bonds and halogen...halogen interactions, the common weak directional interactions in soft organic matter. Pressure-frozen chloroiodomethane, CH(2)ClI, at 295 K and 0.72 GPa forms centrosymmetric phase III, which at ca. 400 K and 1.6 GPa disproportionates into CH(2)Cl(2) and CH(2)I(2). The directional character of intermolecular contacts between halogen atoms results from the characteristic anisotropic charge distribution on molecular surface. PMID- 18402489 TI - Programmed metalloporphyrins for self-assembly within light-harvesting stacks: (5,15-dicyano-10,20-bis(3,5-di-tert-butylphenyl)porphyrinato)zinc(II) and its push-pull 15-N,N-dialkylamino-5-cyano congeners obtained by a facile direct amination. AB - The title dicyano compound was synthesized via cyanation and it self-assembles in nonpolar solvents giving red-shifted and broad absorption maxima just as the bacteriochlorophylls which are encountered in the light-harvesting organelles of early photosynthetic bacteria. In the crystal, stacks are formed through a hierarchic combination of pi-stacking and a CN-Zn electrostatic interaction. Push pull 15-N,N-dialkylamino-5-cyano congeners could be obtained in high yields using a solvent- and catalyst-free direct amination of meso-bromoporphyrins. Importantly, the fluorescence of the self-assembled species due to the very orderly manner in which the chromophores are arranged is not entirely quenched and has a surprisingly long lifetime of over 1 ns. This lends hope of using the trapped energy in biomimetic hybrid solar cells. PMID- 18402490 TI - Supramolecular self-organization in PEO-modified C60 fullerene/water solutions: influence of polymer molecular weight and nanoparticle concentration. AB - Utilizing a first-principles-based coarse-grained implicit solvent model, we have investigated the self-association of C(60) fullerenes that have been symmetrically modified with six grafted poly(ethylene oxide) (PEO) chains in aqueous solution. Despite the highly symmetric nature of the pair interactions between PEO-grafted fullerenes, their supramolecular assemblies are highly anisotropic and resemble the linear clusters formed in Stockmayer fluids. The dipole-like interaction between these symmetrically modified fullerenes results from the shielding of the C(60) fullerenes by PEO, favoring the addition of more PEO-grafted fullerenes to the linear clusters at the relatively unprotected ends. At low nanoparticle concentrations, self-association is dominated by the formation of stable dimers and trimers resulting from fullerene-fullerene contact and favorable PEO-fullerene interactions. With increasing nanoparticle concentration, larger clusters become increasingly probable. The molecular weight of the PEO tethers can be treated as a temperature-like analogue, with a reduction in average cluster size with increasing chain length due to increased steric repulsion, which is qualitatively similar to effects observed in Stockmayer fluids with increasing temperature. The role of PEO in supramolecular self-organization in PEO-modified C(60) fullerene/water solutions is complex, contributing not only to steric stabilization but also to favorable energetic interactions, nanoparticle shielding, and depletion-driven aggregation. PMID- 18402491 TI - Radiation-induced synthesis of gold nanoparticles within lamellar phases. Formation of aligned colloidal gold by radiolysis. AB - A sheared lamellar phase has been used as a nanoreactor for the synthesis of gold nanoparticles by radiolysis and by a photochemical approach. A gold salt solution (KAuCl4, 10(-2) M) is introduced into the aqueous compartments between the lipid based bilayers. Gold nanoparticles grow within the lamellar phase as shown by TEM analysis and X-ray diffraction, limiting the particle size. Homodisperse, 2.4- and 5.9-nm-sized, spherical nanoparticles are produced by gamma irradiation and UV irradiation, respectively. When produced by radiolysis, they are perfectly aligned along the lamellae. Their UV-vis spectra display a maximum at 565 nm suggesting that nanoparticles are coupled by dipole-dipole interactions within the lamellar phase. PMID- 18402492 TI - Supramolecular self-assembled arrangements of maltose glyconanoparticles. AB - Our group previously reported the preparation of water-soluble Au-Fe(x)O(y) nanoparticles functionalized with a maltose neoglycoconjugate. A fraction soluble in methanol was also separated and originated a new supramolecular polymeric aggregate. We report here the full characterization of this novel material by transmission electron microscopy (TEM), fluorescence emission, and atomic force microscopy. By means of noncontact dynamic atomic force microscopy, we have been able to obtain information about the organization of the organic components of the polymers, which eluded TEM analysis. We have observed that polymers packed in units about 65 nm in length and 40 nm in width on Au surfaces. The nanoparticles seem to be encapsulated by the organic material. We propose interactions between the sugar residues and the amphiphilic character of the maltose neoglycoconjugate (with a lipophilic undecane spacer) as responsible for the origin of these amazing supramolecular arrangements. PMID- 18402493 TI - Mixing in adsorbed monolayers: perfluorinated alkanes. AB - The mixing behavior of binary combinations of perfluoroalkanes in the bulk and in solid monolayers adsorbed at the graphite/liquid interface, determined by calorimetry and powder diffraction, is reported. The perfluoroalkanes are found to generally have a smaller excess enthalpy of mixing on the surface than in the bulk, and their relative size ratio is a good parameter to predict the mixing behavior. The excess enthalpy of mixing for perfluoroalkanes is found to be significantly smaller than that of the closely related hydrocarbons. The preferential adsorption of longer homologues over shorter ones is observed. Interestingly, the extent of preferential adsorption with relative size ratio is very similar to that of the hydrocarbons. These results can be understood in terms of the increased compressibility and lower polarizability of the perfluoroalkanes compared to hydrocarbons. PMID- 18402494 TI - Managing bipolar disorder from urgent situations to maintenance therapy. AB - Treatment for bipolar disorder consists of 2 goals: 1) managing urgent situations and 2) transitioning patients into maintenance therapy. Patients with bipolar disorder who present in urgent situations, including symptoms of suicidality, aggression, occupational impairment, legal involvement, and psychosis, require quick and efficient identification and resolution of symptoms. Once patients are stabilized, clinicians must then implement maintenance therapy to manage behavioral symptoms, promote optimal functioning, and ultimately, to reach full, functional recovery. PMID- 18402495 TI - Management of patients with treatment-resistant depression. AB - Treatment-resistant depression can be attributed to multiple causes, including nonadherence to medication and medical and psychiatric comorbidities. Selecting the appropriate strategy for a patient with treatment-resistant depression, whether it be a single antidepressant, a combination of medications, or a nonpharmacologic method, depends on a correct determination of the cause of the patient's resistance to treatment. PMID- 18402496 TI - The bipolar spectrum in children and adolescents: developmental issues. AB - Bipolar disorders are common, chronic illnesses that can develop in children and adolescents at early ages. However, diagnosing bipolar disorders in youths can be difficult because currently defined gradations of the disorder, including bipolar I and bipolar II disorders and bipolar disorder not otherwise specified, were not created with the developmental differences of children in mind. Children with major depression or attention-deficit/hyperactivity disorder plus a family history of bipolar disorder may be presenting with prodromal signs and symptoms of a bipolar disorder, although longitudinal studies are needed to clarify risk factors for developing bipolar disorders. Neuropsychological and biological markers may eventually aid clinicians in distinguishing bipolar spectrum disorders and offering early intervention for at-risk children and adolescents. PMID- 18402497 TI - Psychosis and agitation in dementia: should general psychiatrists care? PMID- 18402498 TI - Clinical trials and tribulations: perspectives on research career development. PMID- 18402499 TI - Clinical insights into pharmacogenetics and schizophrenia, part 1. PMID- 18402500 TI - What constitutes evidence-based pharmacotherapy for bipolar disorder? Part 2: complex presentations and clinical context. PMID- 18402501 TI - Quetiapine treatment for mania secondary to brain injury in 2 patients. PMID- 18402502 TI - Gender imbalance in clinical trials in schizophrenia. PMID- 18402504 TI - Misdiagnosis of bipolar II disorder as major depressive disorder. PMID- 18402503 TI - Urinary retention associated with ziprasidone: a case report. PMID- 18402505 TI - Treating schizophrenia with comorbid depressive or demoralization symptoms. PMID- 18402507 TI - Parkinson's disease--Part 1: Pathophysiology, symptoms, burden, diagnosis, and assessment. AB - Parkinson's disease (PD) is a chronic neurodegenerative disease associated with substantial morbidity, increased mortality, and high economic burden. Of importance to managed care is that the number of cases of PD are on the rise, paralleling the advancing age of the population, and misdiagnosis is common. Effective management of PD can minimize disability and potentially improve long term outcomes, which would minimize long-term healthcare costs and medical resource utilization. This article provides a brief review of the epidemiology, pathophysiology, clinical course, and burden of PD. PMID- 18402508 TI - Parkinson's disease--Part 2: Treatment of motor symptoms. AB - In the absence of a cure, the primary goals in managing Parkinson's disease (PD) are to preserve functionality and health-related quality of life. Meeting these goals can minimize healthcare-resource utilization and long-term healthcare costs. Although effective treatment of motor symptoms of the disease is a central consideration to facilitate improved outcomes, management of nonmotor symptoms is now recognized as an equally important target of intervention, since nonmotor symptoms can contribute greatly to disability. The article addresses the current treatment options of choice for reducing motor symptoms of PD and their most rational use. Cost-effectiveness is a major consideration for managed care and is also analyzed for many available treatment options. PMID- 18402509 TI - Parkinson's disease--Part 3: Neuropsychiatric symptoms. AB - The nonmotor neuropsychiatric symptoms of Parkinson's disease, particularly depression, psychosis, and cognitive impairment/dementia, are major contributors to disability and a decline in quality of life. Their effect on patients may be more disabling than motor symptoms. Increasing awareness of the importance of recognizing and treating neuropsychiatric symptoms of this disease in the medical community is a focus of specialists and organizations. This article looks at useful screening measures to help clinicians recognize neuropsychiatric symptoms and offers suggestions for their effective treatment. PMID- 18402510 TI - A strategic plan for integrating cost-effectiveness analysis into the US healthcare system. AB - The Panel on Integrating Cost-Effectiveness Considerations into Health Policy Decisions, composed of medical and pharmacy directors at public and private health plans, was convened to (1) explore the views of health plan purchasers about cost-effectiveness analysis (CEA) and (2) to develop a strategic plan for policymakers to address obstacles and to integrate CEA into health policy decisions, drawing on stakeholders as part of the solution. Panelists expressed strong support for a greater role for CEA in US health policy decisions, although they also highlighted barriers in the current system and challenges involved in moving forward. The strategic plan involves a series of activities to advance the use of CEA in the United States, including research and demonstration projects to illustrate potential gains from using the technique and ongoing consensus- building steps (eg, workshops, conferences, town meetings) involving a broad coalition of stakeholders. Funding and leadership from policymakers and nonprofit foundations will be needed, as well as the active engagement of legislators and business and consumer groups. Panelists emphasized the importance of the Medicare program taking a lead role, and the need for new "infrastructure," in the form of either a new institute for conducting research or increased funding for existing institutions. PMID- 18402511 TI - Costs of treating lower respiratory tract infections. AB - OBJECTIVE: To determine the direct medical costs of treating lower respiratory tract infections (LRTIs) in a managed care organization (MCO). STUDY DESIGN: Retrospective analysis of a regional MCO identifying adults diagnosed with acute exacerbation of chronic bronchitis (AECB) or community- acquired pneumonia (CAP). METHODS: A claims database examination of International Classification of Diseases, Ninth Revision, Clinical Modification codes was conducted to identify adults receiving initial outpatient care for an LRTI during 2005-2006. Medical record review then was conducted to verify clinical diagnosis of AECB or CAP. Clinical and demographic data were collected. Outpatient office and clinic visits, hospitalization, and radiology, pathology, and pharmacy records were used to determine treatment costs. Treatment failure was determined by use of a second antibiotic course, follow-up emergency room presentation, or hospitalization for LRTI within 28 days of the index visit. The primary outcome was per-case treatment cost from the payer perspective. RESULTS: Clinical diagnosis was confirmed for 65 unique coded visits (60 patients; 39 with AECB, 22 with CAP; 1 in both cohorts). Initial visit, initial diagnostic testing, and subsequent hospitalization accounted for the majority (63%) of payer costs. Antibiotics were responsible for 15% of payer costs. Higher initial antibiotic expenditure in the AECB cohort yielded a cost-benefit ratio of 3:1. Mean per-case costs for success and failure were $277 & $372 for AECB, and $493 & $3019 for CAP, respectively. CONCLUSIONS: Initial visit and hospitalization costs contribute the majority of payer expenditure while antibiotic expenditure incurs a nominal burden. Higher expenditure on initial antibiotic therapy in the AECB population appears to be beneficial. PMID- 18402512 TI - Association of insurance coverage with chlamydia screening. AB - OBJECTIVES: To examine the rates of self-reported Chlamydia trachomatis (CT) screening among young women and to examine the independent association of type of insurance and specific health plans with these rates. STUDY DESIGN: Cross sectional analyses of the 2003 California Health Interview Survey data. METHODS: Using bivariate analysis and logistic regression models, we assessed the CT screening rate of 1659 sexually active women age 18-25 years, given various factors including type of health insurance coverage. We further assessed the CT screening rate of the subset of 533 sexually active women age 18-25 years enrolled in a private health plan and reexamined the relationship of various factors with CT screening rates. RESULTS: Being older, an immigrant, or having 1 sexual partner reduced the likelihood of CT screening, while being a smoker, being single, or having had multiple doctor visits as well as a Pap test or clinical breast exam increased this likelihood. The uninsured had the lowest rate, and public managed care enrollees had the highest rate, of CT screening, but this insurance effect was superseded by other explanatory variables. A few differences in significantly associated factors were identified when private health plans were separately examined. CONCLUSIONS: The results suggest that self reported CT screening rates were low, particularly among the uninsured. However, these rates were primarily influenced by CT risk factors rather than insurance coverage. Continued efforts to increase CT screening rates are warranted. PMID- 18402513 TI - Asthma-specific quality of life and subsequent asthma emergency hospital care. AB - OBJECTIVE: To identify an optimal cut-point score on the Mini-Asthma Quality of Life Questionnaire (mini-AQLQ) to predict subsequent asthma exacerbations, and to determine the additional risk conferred by a prior history of acute episodes. STUDY DESIGN: Cross-sectional survey linked to administrative records. METHODS: A total of 1006 HMO patients with active asthma completed surveys that included the mini-AQLQ and prior-year history of acute episodes. Surveys were linked to administrative data that captured asthma emergency department and hospital care (emergency hospital care) for the year after the survey. Optimal mini-AQLQ cut point scores were determined by stepwise logistic regression analyses using subsequentyear asthma emergency hospital care as the outcome and various mini AQLQ cut-points as the predictors. Predictive properties of the 2 risk factors (mini-AQLQ cut-points and prior acute episodes) were determined. RESULTS: A mini AQLQ cut-point of 4.7 was most significantly associated with subsequent exacerbations in patients without a history of prior acute episodes. The presence of either a mini-AQLQ score <4.7 or a history of prior acute episodes provided high sensitivity (90.4%) and identified a group nearly 6 times more likely to require emergency hospital care than patients with neither risk factor. The presence of both risk factors provided high specificity (79.2%) and resulted in a risk ratio of 9.5 compared with the absence of both risk factors. CONCLUSION: Asthma-specific quality of life and a history of acute episodes can be used together to identify patients with clinically meaningful higher and lower risks of subsequent acute exacerbations. PMID- 18402514 TI - Does sharing care across the primary-specialty interface improve outcomes in chronic disease? A systematic review. AB - OBJECTIVE: To determine the effectiveness of shared care interventions designed to improve the management of chronic disease across the primary-specialty care interface. STUDY DESIGN AND METHODS: Systematic review using the Cochrane Collaboration method. RESULTS: Twenty studies were identified, 19 of which were randomized controlled trials. The majority of studies examined complex interventions and were of short duration. Results were mixed, with no consistent improvements in physical or mental health outcomes, psychosocial outcomes, hospital admissions, default or participation rates, recording of risk factors, and satisfaction with treatment. However, there were improvements in prescribing in the studies that considered this outcome. The methodologic quality of studies varied, with only a minority of studies of high-quality design. Cost data were limited and difficult to interpret across studies. CONCLUSIONS: At present, there is insufficient evidence to support the introduction of shared care services into clinical practice. However, methodologic shortcomings, particularly inadequate length of follow-up, may account for this lack of evidence. Further research is needed to test models of collaboration across the primary-specialty care divide both in terms of effectiveness and sustainability over longer periods of time. PMID- 18402515 TI - Characteristics of and trends in the late-stage biopharmaceutical pipeline. AB - OBJECTIVE: To quantify and characterize biopharmaceutical agents and new indications in late-stage development in the United States as of May 2006. STUDY DESIGN: Review of drug development databases and other secondary sources. METHODS: Biopharmaceutical was defined as "any biology-based therapeutic that structurally mimics compounds found within the body." Unique biopharmaceuticals, including new molecular entities or new indications in phase 2 or higher development, were identified and characterized through reviews of the literature, 5 drug development databases, a clinical trial database, and telephone inquiries with manufacturers. RESULTS: As of May 2006, there were 111 unique biopharmaceuticals in late-stage development for 190 indications. Of 111 unique agents in the pipeline, 87 are new molecular entities, and 24 are already approved for other indications. Overall, 38 disease categories were targeted, and at least 33 physician specialties are likely to be affected. The greatest proportion of agents (43 biopharmaceuticals and 83 indications) target cancer. More than 70% of agents in the pipeline will require administration by a healthcare provider. More than 50% of the indications in the pipeline will require long-term (chronic) treatment (defined as >1 year and excludes cancer). CONCLUSIONS: The steady growth of the US biopharmaceutical pipeline and consequent anticipated near-term approvals will increasingly affect third-party portfolio decision making. Cost of therapy, identifying the right drug for the right patient, and outcomes-based value should drive that decision process. PMID- 18402516 TI - Successful targeted therapies for hepatocellular carcinoma: are we really getting there? PMID- 18402517 TI - Potential new drug targets against hormone-dependent breast cancer identified. PMID- 18402518 TI - Gemcitabine in the treatment of metastatic pancreatic cancer. AB - Gemcitabine (2 ,2 -difluorodeoxycytidine) is a deoxycytidine-analog antimetabolite with broad activity against a variety of solid tumors and lymphoid malignancies. It was approved as standard of care in patients with pancreatic cancer one decade ago, based primarily on improvement in clinical benefit response such as pain reduction, improvement in Karnofsky performance status and increase in body weight. This article gives an overview of the pharmacodynamics and pharmacokinetics of gemcitabine, highlights the clinical activity of gemcitabine and summarizes the treatment options in metastatic pancreatic cancer with focus on gemcitabine-based chemotherapy. The emerging role of combinations of gemcitabine with novel targeted agents, including small-molecule inhibitors and other investigational drugs, is also discussed. PMID- 18402519 TI - Alemtuzumab in the treatment of chronic lymphocytic lymphoma. AB - Alemtuzumab was the first monoclonal antibody to be humanized, a process which embeds rodent sequence fragments in a human IgG framework. The antibody target is CD52, an antigen expressed on normal lymphocytes as well as many T- and B-cell neoplasms. It therefore has a potential broad application across a spectrum of B- and T-cell malignancies as well as use as an immunosuppressant drug in, for example, bone marrow transplantation. The original licensing in the USA and Europe was for the treatment of fludarabine-refractory chronic lymphocytic leukemia (CLL). However, recent trials using alemtuzumab as a first-line agent for CLL have shown superior response rates compared with traditional alkylator therapy and this has led to US FDA approval for first-line treatment for CLL. It seems to be particularly useful in patients with CLL who have deletion of the TP53 tumor suppressor gene, a subset of disease that responds poorly to other currently available chemotherapeutics. PMID- 18402520 TI - Cladribine in indolent non-Hodgkin's lymphoma. AB - Before the advent of rationally designed targeted antineoplastic therapies, cladribine was identified as a lymphocyte-specific cytotoxic agent. Cladribine is a purine nucleoside analogue that is resistant to cellular catabolism. Through diverse mechanisms, cladribine is equally toxic to dividing and nondividing cells, making it highly active in indolent lymphoproliferative diseases. In clinical practice, cladribine is mostly used in the treatment of hairy cell leukemia and Waldenstrom's macroglobulinemia. However, its remarkable activity in follicular lymphoma and other indolent non-Hodgkin's lymphoma subtypes has not been more widely appreciated. Cladribine compares favorably to other standard treatments for these conditions. Future Phase III clinical studies should incorporate cladribine into multiagent chemotherapy programs to more fully evaluate its potential in indolent non-Hodgkin's lymphoma. PMID- 18402521 TI - Redefining adjuvant chemotherapy in patients with stage III colon cancer: X-ACT trial. AB - The current standard adjuvant chemotherapy for suitable patients with stage III colon cancer is the combination of oxaliplatin and 5-fluorouracil plus folinic acid (5-FU/LV). However, until recently and for many years prior to this, the accepted standard adjuvant chemotherapy was 6-8 months of bolus 5-FU/LV. However, bolus treatment was associated with significant toxicity, namely stomatitis, diarrhea and neutropenia, in addition to multiple hospital visits for drug administration for patients. The X-ACT trial (Xeloda in Adjuvant Colon Cancer Therapy) compared traditional bolus 5-FU/LV (as per the Mayo Clinic regimen) with capecitabine, in the adjuvant treatment of 1987 stage III colon cancer patients. The main safety, efficacy and pharmacoeconomic results have all been published, and the updated 5-year efficacy results have also recently been presented. This trial demonstrated that capecitabine was at least as effective as bolus 5-FU/LV in terms of disease-free and overall survival, with trends towards superiority for both. Moreover, there was much less toxicity associated with capecitabine, apart from hand-foot syndrome which was significantly more prevalent. On the basis of the X-ACT trial, capecitabine was approved by the US FDA, the National Institute for Clinical Excellence and the Scottish Medicines Consortium as monotherapy for the adjuvant treatment of stage III colon cancer. PMID- 18402522 TI - Emerging therapies for melanoma. AB - Substantial progress has been made in understanding the pathogenesis of malignant melanoma at the cellular level, leading to the development of several new agents in the treatment of melanoma. The focus of this review is to summarize the emerging therapies and investigational agents in the treatment of melanoma over the last 5 years. PMID- 18402523 TI - Immune cells in colorectal cancer: prognostic relevance and therapeutic strategies. AB - During the last two decades, considerable efforts have been made to improve the prevention, early diagnosis and therapy of colorectal cancer by gaining enhanced insights into disease-specific pathogenesis. Along these lines, tumor infiltrating immune cells turned out to be critical indicators for an efficient antitumor immune response and the number and type of tumor-infiltrating immune cells determined the resulting tumor prognosis. This review aims to describe the prognostic relevance of the different subsets of tumor-infiltrating immune cells and highlights their specific function in the complex process of immune system mediated rejection of colorectal cancer cells. Considering the clinical impact of this increased insight into tumor-associated immunology, resulting innovative approaches in the treatment of colorectal cancer are summarized and discussed immunotherapeutic critically. PMID- 18402524 TI - Lynch syndrome in colorectal cancer patients. AB - Lynch syndrome, also known as hereditary nonpolyposis colorectal cancer, is the most common form of hereditary colorectal cancer. It is characterized by early onset of colorectal cancer and other extracolonic-associated malignancies. This disorder is inherited in an autosomal dominant pattern and is due to a mutation in one of the DNA mismatch repair genes. Although clinical and molecular understanding of the syndrome has progressed dramatically in the last decade, diagnosis of the syndrome is still a clinical challenge. This review summarizes the main features of the syndrome and provides an update of its management. PMID- 18402525 TI - Emerging role of intensity-modulated radiation therapy in anorectal cancer. AB - Although radiation therapy has an established role to play in the management of rectal and anal tumors, there are often treatment-related morbidities that negatively impact on patients. There is a long-standing interest in radiation oncology on maximizing treatment efficacy while minimizing treatment-related toxicities, which can be pronounced in the treatment of pelvic malignancies. Intensity-modulated radiation therapy is a recently introduced technology that has the potential to increase the efficacy:toxicity ratio. It has been implemented in the treatment of prostate and head and neck tumors with success. This article reviews the rationale for its use in treating anorectal tumors and discusses early clinical data supporting its continued investigation. PMID- 18402526 TI - Progress and challenges in the adjuvant treatment of stage II and III colon cancers. AB - Whereas the benefit of adjuvant 5-fluorouracil and leucovorin have been well established in resected stage III colon cancer, a significant benefit for patients with stage II disease has been more difficult to demonstrate. More recently, oxaliplatin-based chemotherapy with regimens such as oxaplatin plus 5 fluorouracil/leucovorin have been shown to improve disease-free and overall survival in these stage III patients. This review will discuss the development of adjuvant chemotherapy in colon cancer, focusing on recent progress and particular topical issues related to its use in this disease, such as the use of surrogate end points for overall survival in contemporary clinical trials. PMID- 18402527 TI - Inflammation in the development of lung cancer: epidemiological evidence. AB - The lung is a site for repeated or chronic inflammatory insults. Epidemiologic research has provided evidence to support the hypothesis that tissue damage caused by inflammation can initiate or promote the development of lung cancer, possibly in conjunction with tobacco use. For example, some studies suggest an increased risk of lung cancer among persons with lung infections, such as tuberculosis, bacterial pneumonia, or inflammatory lung diseases. Elevated serum levels of C-reactive protein, an inflammation marker, are associated with heightened lung cancer risk. Recent studies also demonstrate increased lung cancer risk among immunosuppressed individuals infected with HIV. Other research indicates an association between genetic polymorphisms in the inflammation pathway, which might modulate the inflammatory response and lung cancer risk. PMID- 18402528 TI - Ewing sarcoma: prognostic criteria, outcomes and future treatment. AB - Ewing sarcoma (EWS) is a bone tumor occurring primarily in adolescence and young adulthood. Multi-institutional clinical trials have improved the outcome for patients with nonmetastatic EWS, but not with metastatic EWS. Furthermore, although 30% of EWS recur, multi-institutional studies have not been completed for this high-risk group. Planning such studies has been hampered by both the lack of novel therapies and the inability to incorporate the biology of EWS. While the importance and detail of the EWS-FLI-1 translocation between chromosomes 11 and 22 are described, these have not yet led to new drug development for this orphan tumor. However, recent evidence supporting novel cytotoxic therapy, antiangiogenic therapy, and receptor-targeted therapy provides reason for optimism for patients with high-risk disease. PMID- 18402530 TI - Advances in radiation therapy of head and neck cancer. AB - Head and neck cancer is the fifth most common cancer in the USA. Although there have been major improvements in surgical and radiation techniques, the overall survival has not changed significantly in the last decade. The major changes occurring in recent years have been in the ability to preserve organs and to improve quality of life. The advances in radiation therapy include 3D conformal radiotherapy, intensity-modulated radiotherapy and, more recently, imaging-guided radiotherapy. In advanced head and neck cancer the addition of chemotherapy to radiation has concomitantly improved survival and facilitated higher rates of organ preservation. Clinical trials are needed to develop better strategies customized to subgroups defined by individual biological risk and imaging findings. PMID- 18402529 TI - RET oncogene in MEN2, MEN2B, MTC and other forms of thyroid cancer. AB - Hereditary medullary thyroid carcinoma (MTC) is caused by specific autosomal dominant gain-of-function mutations in the RET proto-oncogene. Genotype-phenotype correlations exist that help predict the presence of other associated endocrine neoplasms as well as the timing of thyroid cancer development. MTC represents a promising model for targeted cancer therapy, as the oncogenic event responsible for initiating malignancy has been well characterized. The RET proto-oncogene has become the target for molecularly designed drug therapy. Tyrosine kinase inhibitors targeting activated RET are currently in clinical trials for the treatment of patients with MTC. This review will provide a brief overview of MTC and the associated RET oncogenic mutations, and will summarize the therapies designed to strategically interfere with the pathologic activation of the RET oncogene. PMID- 18402531 TI - Salivary gland cancers: current treatments, molecular characteristics and new therapies. AB - Salivary gland cancers are relatively rare and quite diverse. Current therapy relies on local ablation. There are few large clinical trials or randomized trials to guide treatment, especially for metastatic disease. This article reviews the epidemiology, staging, molecular characteristics, and treatment evidence for the most common types of salivary cancers and suggests potential future diagnostic and treatment directions. Progress in understanding the molecular and cell biology of salivary gland cancers may lead to the development of targeted therapies in these rare tumors. Multidisciplinary and multi institutional collaborative studies are needed to help improve survival in salivary gland cancers. PMID- 18402533 TI - Tailor-made therapy for the prevention of acute coronary syndromes: future role of biomarkers in risk stratification and disease management. PMID- 18402534 TI - Surgical management of congestive heart failure: translational research to clinical application--the future is bright! PMID- 18402536 TI - Ezetimibe: cholesterol lowering and beyond. AB - Ezetimibe is a cholesterol absorption inhibitor that blocks the intestinal absorption of both biliary and dietary cholesterol. It appears to exert its effect by blocking intestinal sterol transporters, specifically Niemann-Pick C1 like 1 proteins, thereby inhibiting the intestinal absorption of cholesterol, phytosterols and certain oxysterols. Ezetimibe monotherapy and in combination with statin therapy is primarily indicated for lowering LDL-cholesterol levels. In addition, it may favorably affect other parameters that could potentially further reduce atherosclerotic coronary heart disease risk, such as raising HDL cholesterol and lowering levels of triglycerides, non-HDL-cholesterol, apolipoprotein B and remnant-like particle cholesterol. Further effects of ezetimibe include a reduction in circulating phytosterols and oxysterols and, when used in combination with statins, a reduction in high-sensitivity C-reactive protein. The clinical significance of the LDL-cholesterol lowering and other effects of ezetimibe is being evaluated in clinical outcome studies. PMID- 18402537 TI - Beta-blockers in the management of hypertension: focus on nebivolol. AB - Hypertension is a major cardiovascular risk factor but most patients remain asymptomatic for many years. Successful therapy not only needs to be effective, it also needs to be well tolerated. beta-blockers are well established as effective antihypertensive agents. However, one major drawback to the currently available beta-blockers, particularly the noncardioselective beta-blockers, is their side-effect profile, including sexual dysfunction, fatigue, depression and metabolic abnormalities such as impaired glucose tolerance and lipid abnormalities. Nebivolol (Bystolic), a novel, highly cardioselective, third generation beta-blocker that recently received approval by the US FDA for the treatment of hypertension in the USA, is effective in treating blood pressure and has a favorable side-effect profile. Studies conducted in Europe, where nebivolol has been available for some time for the treatment of hypertension, have shown that nebivolol achieves blood pressure reductions comparable to other beta blockers but with fewer side effects. Additionally, nebivolol has demonstrated similar efficacy in blood pressure reduction when compared with calcium channel blockers and inhibitors of the renin-angiotensin system. When combined with hydrochlorothiazide there was an additive antihypertensive effect. Lastly, nebivolol exhibits a vasodilatory property that is related to its effect on nitric oxide, an intrinsic vasodilator produced in the vascular endothelium. Nebivolol enhances nitric oxide bioavailability. Studies have also demonstrated nebivolol's ability to function as an antioxidant and decrease markers of oxidative stress. These effects are believed to ultimately produce a modulation of the endothelial dysfunction typically seen in hypertension. PMID- 18402538 TI - Current surgical management of mitral regurgitation. AB - From Walton Lillehei, who performed the first successful open mitral valve surgery in 1956, until the advent of robotic surgery in the 21st Century, only 50 years have passed. The introduction of the first heart valve prosthesis, in 1960, was the next major step forward. However, correction of mitral disease by valvuloplasty results in better survival and ventricular performance than mitral valve replacement. However, the European Heart Survey demonstrated that only 40% of the valves are repaired. The standard procedures (Carpentier's techniques and Alfieri's edge-to-edge suture) are the surgical basis for the new technical approaches. Minimally invasive surgery led to the development of video-assisted and robotic surgery and interventional cardiology is already making the first steps on endovascular procedures, using the classical concepts in highly differentiated approaches. Correction of mitral regurgitation is a complex field that is still growing, whereas classic surgery is still under debate as the new era arises. PMID- 18402539 TI - Optimum management of elderly patients with calcified aortic stenosis. AB - Increased life-expectancy has led to a growing elderly population frequently presenting with aortic stenosis. This review focuses on the pathogenesis of calcific aortic stenosis, diagnosis and possible ways to halt the progression to severe symptomatic aortic stenosis, methods of assessing symptoms and severity, and modalities and timing of aortic valve replacement. At present the treatment of aortic stenosis for the majority of patients is surgical, and any patient with symptomatic severe aortic stenosis should be considered for aortic valve replacement. This article also discusses the role of emerging techniques of closed heart valve implantation either transfemoral or transapical, and which patients might be candidates for these new approaches to the treatment of aortic stenosis in the elderly population. PMID- 18402540 TI - Neuroprotection during cardiac surgery. AB - Cardiac surgery continues to be associated with significant adverse cerebral outcomes, ranging from stroke to cognitive decline. The underlying mechanism of the associated cerebral injury is incompletely understood but is believed to be primarily caused by cerebral embolism and hypoperfusion, exacerbated by ischemia/reperfusion injury. Extensive research has been undertaken in an attempt to minimize the incidence of perioperative cerebral injury, and both pharmacological and nonpharmacological strategies have been investigated. Although many agents demonstrated promise in preclinical studies, there is currently insufficient evidence from clinical trials to recommend the routine administration of any pharmacological agents for neuroprotection during cardiac surgery. The nonpharmacological strategies that can be recommended on the basis of evidence include transesophageal echocardiography and epiaortic ultrasound guided assessment of the atheromatous ascending aorta with appropriate modification of cannulation, clamping or anastomotic technique and optimal temperature management. Large-scale randomized controlled trials are still required to address further the issues of optimal pH management, glycemic control, blood pressure management and hematocrit during cardiopulmonary bypass. Past, present and future directions in the field of neuroprotection in cardiac surgery will be discussed. PMID- 18402541 TI - New strategies for surgical management of ischemic cardiomyopathy. AB - Ischemic cardiomyopathy affects an estimated 3 million people in the USA and is the most common cause of heart failure. Traditional operations have included heart transplantation, myocardial revascularization, mitral valve repair, left ventricular reconstruction, first-generation left ventricular assist devices and cardiac resynchronization therapy. These operations have become safer in recent times, due to improved technologies. Current technologies and surgical approaches can benefit a significant number of patients. However, there remains a large group of patients in whom traditional approaches can not be offered. Newer generation ventricular assist devices, passive ventricular restraint devices and cellular-based therapies (including skeletal- and bone marrow-derived stem cells) have the potential to be more universal in their applications. Ongoing investigations with each of these modalities will allow surgeons to offer treatment to patients who are not considered surgical candidates at this time. PMID- 18402542 TI - Current status of bare and drug-eluting stents in infrainguinal peripheral vascular disease. AB - Currently, the use of endovascular therapies including stent application is increasingly becoming the treatment of choice for superficial femoral and below the-knee lesions. Primary stenting of even longer lesions is supported by favorable results of mid- to long-term evaluations. However, issues such as the long-term patency of these devices, optimal design of the stents used, and the role of coating with drugs for infrainguinal disease are under investigation. With more widespread use, problems such as fractures become evident and further developments will have to address these issues. Currently, a number of studies are being conducted, which will help to clarify the role of patient and lesion selection, choice of device, design of the stent and the question of whether or not to use drug-eluting stents for this application. Furthermore, endovascular therapy of below-the-knee lesions is becoming an increasingly interesting area for interventional procedures. PMID- 18402543 TI - Peripheral artery disease: therapeutic advances. AB - Peripheral arterial disease (PAD), usually caused by atherosclerosis, is defined as an obstructive arterial disease of the lower extremities that reduces arterial flow during exercise or, in advanced stages, at rest. It affects more than 8.5 million people in the USA. PAD may appear as an asymptomatic arterial disease with abnormal noninvasive test results, or as a symptomatic disease presenting with atypical limb pain, classic intermittent claudication, or critical limb ischemia. The spectrum of PAD is not a continuum. Patients who present with critical limb ischemia may have experienced minimum symptoms. PAD results in limitation of exercise and walking ability, described as intermittent claudication. Patients with PAD are physically impaired and have a higher risk of cardiovascular events; therefore, the treatment goals are aimed at decreasing their cardiovascular risk, as well as improving exercise and daily functional performance. Apart from supervised exercise, which is a major treatment modality for patients with PAD, as of yet there have been very few significant pharmacological breakthroughs in the treatment of PAD that increases blood flow to the ischemic limb. Although percutaneous intervention has markedly improved the treatment of PAD, bypass surgery continues to play an important role. For the most part medical therapy for PAD is designed as a secondary prevention for cardiovascular risk. These include antiplatelet therapy, statins, ACE-inhibitors, smoking cessation and possibly antihypertensive therapy. Revascularization is most beneficial for patients with lifestyle limiting symptoms, acute or chronic limb ischemia with resting pain or nonhealing ulcers. In the following review article we will try to explore the clinical role of some of the latest developments in this field. PMID- 18402544 TI - Sex differences in response to chronic heart failure therapies. AB - Sex-related differences in clinical and laboratory characteristics, course and prognosis are well documented in patients with heart failure. However, most information regarding heart failure therapies has been obtained from studies conducted primarily in men. Reviewing the existing literature indicates that the recommendations regarding pharmacological and device therapies should apply similarly to men and women. One possible exception, however, is the possibility of more benefit derived from angiotensin receptor blockers in women, and from angiotensin converting enzyme inhibitors in men. Future clinical trials should be conducted either exclusively in women or have a larger representation of women to insure the adequate assessment of the potential benefit versus risk in women. PMID- 18402545 TI - Familial hypercholesterolemia: current treatment and advances in management. AB - Heterozygous familial hypercholesterolemia is associated with elevated levels of LDL-cholesterol and the development of premature cardiovascular disease. The condition is considerably under-diagnosed and under-treated. Statins are the first choice treatment for all patients with heterozygous familial hypercholesterolemia. For those patients who do not reach their treatment target or who are unable to use adequate statin dose, several alternative treatment modalities can be used, either as add-on therapy or as monotherapy. In this review the various treatment options are discussed. PMID- 18402549 TI - Cattle density and Shiga toxin-producing Escherichia coli infection in Germany: increased risk for most but not all serogroups. AB - Shiga toxin-producing Escherichia coli (STEC) cause severe gastroenteritis and life-threatening hemolytic-uremic syndrome. For STEC of serogroup O157, association between disease incidence and cattle contact has been established in some countries. For other (non-O157) serogroups, however, accounting for approximately 80% of notified STEC gastroenteritis in Germany, the role of cattle in human infection is less clear. For example, an association of non-O157 STEC infection and cattle density has not been investigated. The aim of this study was thus to investigate a potential association between STEC incidence and cattle density in Germany, with special attention to the non-O157 serogroups. We modeled district-level incidence of notified human STEC cases in relation to cattle density, utilizing German notification data from 2001 through 2003. Cattle numbers came from the national "Proof of Origin and Information Database for Animals." A Bayesian Poisson regression model was used, incorporating independent, as well as spatially correlated, district-level random effects into the analysis. We analyzed 3216 German STEC cases. Cattle density was positively associated with overall STEC incidence. The risk for STEC infection increased by 68% per 100 additional cattle/km(2). The magnitude of the risk estimates differed by serogroup and was greatest for O111. A positive association was found for all major disease-causing serogroups (O26, O103, O111, O128, O145, O157) except O91. The association with serogroup O26 (lowest median age of patients) was only borderline significant. Residual variation indicates that additional factors not under study may also be of importance, and that they may be serogroup- and region specific, too. In conclusion, this study suggests that living in a cattle-raising region appears to imply risk not only for STEC O157, but also for most non-O157 serogroups. Furthermore, the varying magnitude of this risk and the residual variation found for different serogroups indicate that risk profiles for human STEC infection may be serogroup-specific. This needs to be taken into account in risk factor studies for non-O157 STEC, ideally by reporting risks separately by serogroup. PMID- 18402550 TI - Redox considerations in female reproductive function and assisted reproduction: from molecular mechanisms to health implications. AB - Physiological levels of reactive oxygen species (ROS) play an important regulatory role through various signaling transduction pathways in folliculogenesis, oocyte maturation, endometrial cycle, luteolysis, implantation, embryogenesis, and pregnancy. Persistent and elevated generation of ROS leads to a disturbance of redox potential that in turn causes oxidative stress (OS). Our literature review captures the role of ROS in modulating a range of physiological functions and pathological processes affecting the female reproductive life span and even thereafter (i.e., menopause). The role of OS in female reproduction is becoming increasingly important, as recent evidence suggest that it plays a part in conditions such as polycystic ovarian disease, endometriosis, spontaneous abortions, preeclampsia, hydatidiform mole, embryopathies, preterm labor, and intrauterine growth retardation. OS has been implicated in different reproductive scenarios and is detrimental to both natural and assisted fertility. Many extrinsic and intrinsic conditions exist in assisted reproduction settings that can be tailored to reduce the toxic effects of ROS. Laboratory personnel should avoid procedures that are known to be deleterious, especially when safer procedures that can prevent OS are available. Although antioxidants such as folate, zinc, and thiols may help enhance fertility, the available data are contentious and must be evaluated in controlled studies with larger populations. PMID- 18402547 TI - Delivery of recombinant adeno-associated virus-mediated human tissue kallikrein for therapy of chronic renal failure in rats. AB - The tissue kallikrein-kinin system is important in regulating cardiovascular and renal function, and dysregulation of the system has been implicated in heart and kidney pathologies. These findings suggest that if balance can be restored to the kallikrein-kinin axis, then associated disease progression may be attenuated. To test this hypothesis, recombinant adeno-associated virus (rAAV)-mediated human tissue kallikrein (HK) expression was induced in a rodent model of chronic renal failure involving 5/6 nephrectomy (nephrectomy plus 70% reduction of remaining kidney). rAAV-HK treatment attenuated the rise in blood pressure, glomerular sclerosis, and tubulointerstitial injury observed in this model. rAAV-HK treatment also attenuated renal function decline as measured by urinary microalbumin, osmolarity, and cGMP levels. Reverse transcriptase-polymerase chain reaction analysis showed that rAAV-HK-treated rats had higher levels of bradykinin receptor-2 (B(2)R) and dopamine receptor-1 mRNAs. In contrast, angiotensin II receptor-1, endothelin receptor-A, and vasopressin receptor-2 mRNAs were markedly downregulated in kidneys from HK-treated rats. Bradykinin induced similar changes in receptor levels and prevented transforming growth factor-beta(1)-induced tubulointerstitial fibrosis. The effects of bradykinin could be reversed with the B(2)R antagonist HOE-140. Together, these findings suggest that restoration of the kallikrein-kinin system reduces kidney injury and protects renal function in 5/6-nephrectomized rats via changes in the expression and activation of G protein-coupled receptors including B(2)R. PMID- 18402551 TI - Gene-modified mesenchymal stem cells express functionally active nerve growth factor on an engineered poly lactic glycolic acid (PLGA) substrate. AB - Delivery of cellular and/or trophic factors to the site of injury may promote neural repair or regeneration and return of function after peripheral nerve or spinal cord injury. Engineered scaffolds provide a platform to deliver therapeutic cells and neurotrophic molecules. We have genetically engineered mesenchymal stem cells (MSCs) from the green rat (CZ-004 [SD TgN(act-EGFP)OsbCZ 004]) to express nerve growth factor (NGF) using an adenoviral vector. Cells maintained their stem cell phenotype as judged by expression of CD71 and CD172 markers, and absence of the hematopoietic marker CD45. Cells continued to express green fluorescent protein (GFP) on a long-term basis. Morphology, viability, and growth kinetics were maintained when cells were grown on a poly-lactic-co glycolic acid (PLGA) polymer scaffold. Under appropriate growth conditions, they differentiated into chondrogenic, osteogenic, and adipogenic phenotypes, demonstrating that they retained their characteristics as MSCs. NGF was secreted from transduced MSCs at physiologically relevant levels ( approximately 25 ng/mL) measured by enzyme-linked immunoabsorbent assay (ELISA). Secreted NGF was functionally active in a neurite growth assay with PC12 cells. We conclude that MSCs are a good candidate for delivery of therapeutic factors into the injured nervous system. They are autologous, may be genetically modified to express neurotrophins, and are compatible with polymer surfaces that may be used as a potential delivery system. PMID- 18402552 TI - Hepatocyte growth factor activator inhibitor-1 has a complex subcellular itinerary. AB - HAI-1 [HGF (hepatocyte growth factor) activator inhibitor-1] is a Kunitz-type transmembrane serine protease inhibitor that forms inhibitor complexes with the trypsin-like serine protease, matriptase. HAI-1 is essential for mouse placental development and embryo survival and together with matriptase it is a key regulator of carcinogenesis. HAI-1 is expressed in polarized epithelial cells, which have the plasma membrane divided by tight junctions into an apical and a basolateral domain. In the present study we show that HAI-1 at steady-state is mainly located on the basolateral membrane of both Madin-Darby canine kidney cells and mammary gland epithelial cells. After biosynthesis, HAI-1 is exocytosed mainly to the basolateral plasma membrane from where 15% of the HAI-1 molecules are proteolytically cleaved and released into the basolateral medium. The remaining membrane-associated HAI-1 is endocytosed and then recycles between the basolateral plasma membrane and endosomes for hours until it is transcytosed to the apical plasma membrane. Minor amounts of HAI-1 present at the apical plasma membrane are proteolytically cleaved and released into the apical medium. Full length membrane-bound HAI-1 has a half-life of 1.5 h and is eventually degraded in the lysosomes, whereas proteolytically released HAI-1 is more stable. HAI-1 is co-localized with its cognate protease, matriptase, at the basolateral plasma membrane. We suggest that HAI-1, in addition to its protease inhibitory function, plays a role in transporting matriptase as a matriptase-HAI-1 complex from the basolateral plama membrane to the apical plasma membrane, as matriptase is known to interact with prostasin, located at the apical plasma membrane. PMID- 18402553 TI - Alternative metabolic fates of phosphatidylinositol produced by phosphatidylinositol synthase isoforms in Arabidopsis thaliana. AB - PtdIns is an important precursor for inositol-containing lipids, including polyphosphoinositides, which have multiple essential functions in eukaryotic cells. It was previously proposed that different regulatory functions of inositol containing lipids may be performed by independent lipid pools; however, it remains unclear how such subcellular pools are established and maintained. In the present paper, a previously uncharacterized Arabidopsis gene product with similarity to the known Arabidopsis PIS (PtdIns synthase), PIS1, is shown to be an active enzyme, PIS2, capable of producing PtdIns in vitro. PIS1 and PIS2 diverged slightly in substrate preferences for CDP-DAG [cytidinediphospho-DAG (diacylglycerol)] species differing in fatty acid composition, PIS2 preferring unsaturated substrates in vitro. Transient expression of fluorescently tagged PIS1 or PIS2 in onion epidermal cells indicates localization of both enzymes in the ER (endoplasmic reticulum) and, possibly, Golgi, as was reported previously for fungal and mammalian homologues. Constitutive ectopic overexpression of PIS1 or PIS2 in Arabidopsis plants resulted in elevated levels of PtdIns in leaves. PIS2-overexpressors additionally exhibited significantly elevated levels of PtdIns(4)P and PtdIns(4,5)P(2), whereas polyphosphoinositides were not elevated in plants overexpressing PIS1. In contrast, PIS1-overexpressors contained significantly elevated levels of DAG and PtdEtn (phosphatidylethanolamine), an effect not observed in plants overexpressing PIS2. Biochemical analysis of transgenic plants with regards to fatty acids associated with relevant lipids indicates that lipids increasing with PIS1 overexpression were enriched in saturated or monounsaturated fatty acids, whereas lipids increasing with PIS2 overexpression, including polyphosphoinositides, contained more unsaturated fatty acids. The results indicate that PtdIns populations originating from different PIS isoforms may enter alternative routes of metabolic conversion, possibly based on specificity and immediate metabolic context of the biosynthetic enzymes. PMID- 18402554 TI - A new biodegradable nanocomposite based on polyhedral oligomeric silsesquioxane nanocages: cytocompatibility and investigation into electrohydrodynamic jet fabrication techniques for tissue-engineered scaffolds. AB - Our group has developed a non-biodegradable nanocomposite based on POSS (polyhedral oligomeric silsesquioxane) nanocages with PCU [poly(carbonate urethane)] and previous studies have shown good cell-compatibility and antithrombogenic properties. The latest biodegradable formulation is a POSS modified poly(hexanolactone/carbonate)urethane/urea containing 80% hexanolactone (caprolactone) with the tradename UCL-NanoBio. The direct effect of the polymer on cells was investigated by seeding stem cells on to circular discs of the polymer in 24-well plates; these discs were prepared mainly by electrohydrodynamic jetting. To assess the indirect effect of the polymer, various concentrations of the polymer powder were added to CCM (cell culture medium) and left on a shaker for 10 days. The precipitate was then removed and the CCM was used for culturing the cells seeded on to 24-well plates. Cell viability and growth at 48 and 96 h were assessed using Alamar Blue and lactate dehydrogenase, and morphology was studied by scanning electron microscopy. Cells were shown to adhere well to the polymer, with cell metabolism being comparable with that found on TCP (tissue-culture plastic). Indirect assessment demonstrated some decrease in cell viability with high concentrations of polymer, but showed no difference in cell death between polymer concentrations. The viability of cells seeded on to the polymer was comparable with that of those seeded on to TCP. Cell viability was comparable on both electrosprayed and electrospun scaffolds, but infiltration into the scaffold was much more evident on the electrospun scaffolds. It can be concluded that this new nanocomposite can support the growth and viability of stem cells and that scaffolds of this polymer nanocomposite fabricated by electrohydrodynamic jetting routes have potential use for tissue engineering in the future. PMID- 18402555 TI - Redox state of quinone affects sensitivity of Acanthamoeba castellanii mitochondrial uncoupling protein to purine nucleotides. AB - We studied FFA (free fatty acid)-induced uncoupling activity in Acanthamoeba castellanii mitochondria in the non-phosphorylating state. Either succinate or external NADH was used as a respiratory substrate to determine the proton conductance curves and the relationships between respiratory rate and the quinone reduction level. Our determinations of the membranous quinone reduction level in non-phosphorylating mitochondria show that activation of UCP (uncoupling protein) activity leads to a PN (purine nucleotide)-sensitive decrease in the quinone redox state. The gradual decrease in the rate of quinone-reducing pathways (using titration of dehydrogenase activities) progressively leads to a full inhibitory effect of GDP on LA (linoleic acid) induced proton conductance. This inhibition cannot be attributed to changes in the membrane potential. Indeed, the lack of GDP inhibitory effect observed when the decrease in respiratory rate is accompanied by an increase in the quinone reduction level (using titration of the quinol-oxidizing pathway) proves that the inhibition by nucleotides can be revealed only for a low quinone redox state. It must be underlined that, in A. castellanii non-phosphorylating mitochondria, the transition of the inhibitory effect of GDP on LA-induced UCP-mediated uncoupling is observed for the same range of quinone reduction levels (between 50% and 40%) as that observed previously for phosphorylating conditions. This observation, drawn from the two different metabolic states of mitochondria, indicates that quinone could affect UCP activity through sensitivity to PNs. PMID- 18402556 TI - DFT-based de novo QSAR of Phenoloxidase Inhibitors. AB - The phenoloxidase or tyrosinase is a key enzyme in insects, which is responsible for hydroxylation of tyrosine into o-quinones via o-diphenols. A series of benzaldehyde thiosemicarbazone, benzaldehyde and benzoic acid families were taken with their pragmatic pIC(50) values against phenoloxidase from pieris rapae (Lepidoptera) larvae. Density functional theory-based quantitative structure activity relationship (QSAR) analyses were performed to speculate the key interaction. The most fitted four different QSAR models were identified and discussed. The softness, electrophilicity index, molar refractivity and log P were identified as best descriptors; however, the atomic values of softness and philicity obtained from Fukui function are more significant than global values. The study reveals that electrostatic and steric fields jointly contribute to activity. To gain further insight, the three-dimensional quantitative structure activity relationship (3D-QSAR) analyses were performed using two molecular field techniques: comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA). The successful 3D-QSAR models were obtained from CoMFA (q(2)= 0.94, r(2)= 0.99, r(2)(pred)= 0.92) and CoMSIA (q(2)= 0.94, r(2)= 0.98, r(2)(pred)= 0/95). The CoMFA and CoMSIA results indicate that, a bulky and negative group around sulfur atom but a small and positive group around nitrogen atom might have good effects on activity. The ortho and meta positions of ring are favorable for small group. These QSAR models might be helpful to design the novel and potent inhibitors. PMID- 18402557 TI - End-of-life decision-making in intensive care: the case for an international standard or a standard of care? PMID- 18402558 TI - Managing high blood glucose levels in coronary care. PMID- 18402559 TI - End of life in the intensive care unit: knowledge and practice of clinicians from Karachi, Pakistan. AB - BACKGROUND: With improvements in the care of critically ill, physicians are faced with obligations to provide quality end-of-life care. Barriers to this include inadequate understanding of the dying patient and withdrawal or limitation of care. The objectives of this study were to document the comprehensions of physicians and nurses regarding the recognition and practice of end-of-life care for critically ill patients placed on life support in the intensive care unit. METHODS: This was a cross-sectional study carried out at three hospitals in Karachi. Chi-squared analysis and one-way ANOVA were used to compare differences in response between the groups. RESULTS: One hundred and thirty-seven physicians and critical care nurses completed the survey. 'Brain death' was defined as an 'irreversible cessation of brainstem function' by 85% respondents, with 50% relying on specialty consultation. Withdrawal of life support is practised by 83.2%; physicians are more likely (Chi square test P-value < 0.001) to withdraw mechanical ventilation, compared with nurses who would withdraw vasopressors (P value 0.006). In a do not resuscitate patient, 72.3% use vasopressors, 83% initiate haemodialysis and 17.5% use non-invasive ventilation; 72.6% consult Hospital Ethics Committees; 16% respondents never withdraw life support; 28.3% considered it their responsibility to 'sustain life at all costs' and only 8% gave religious beliefs as a reason. CONCLUSIONS: There are confusions in the definition of brain death, end-of-life recognition and indications and processes of withdrawal of life support. There are discrepancies between physicians' and nurses' perceptions and attitudes. Clearly, teaching programmes will need to incorporate cultural and religious differences in their ethics curricula. PMID- 18402560 TI - Glucose control peri-myocardial infarction. AB - This review summarises the available clinical trials data investigating the effects of glucose lowering on mortality in patients admitted to hospital with acute myocardial infarction. The results of these studies are inconclusive with no clear evidence that this intervention has additional benefit over and above routine care. PMID- 18402561 TI - Glucose control during acute myocardial infarction. AB - The need for tight glycaemic control in the immediate aftermath of myocardial infarction is controversial. Trials of glucose-insulin-potassium (GIK) therapy, given without regard to glucose levels, have only confused the issue. Despite considerable clinical interest, only three randomized controlled trials have, as their primary goal, aimed to determine whether better glycaemic control improves the outcomes of myocardial infarction. This review examines the results of these trials and other data to support the case for tight glycaemic control in patients with myocardial infarction. PMID- 18402562 TI - Experience with a new commercial skin testing kit to identify IgE-mediated penicillin allergy. AB - Many patients who describe a history of allergy to penicillin do not prove to be allergic and can be treated safely with penicillin. After a period of 2 years where testing of penicillin allergy was not possible, a new commercial kit has recently become available. We report our initial experience with use of the kit with 29 patients and discuss one patient who experienced anaphylaxis during i.d. testing. PMID- 18402563 TI - Medications for the secondary prevention of ischaemic heart disease and heart failure are underutilized in dialysis patients. AB - We determined the prescription rates of medications for the secondary prevention of ischaemic heart disease and left ventricular systolic dysfunction in stable dialysis patients. In patients with established ischaemic heart disease, statins, beta-blockers and renin-angiotensin-aldosterone system blockers were substantially underprescribed. Furthermore, beta-blockers and renin-angiotensin aldosterone system blockers were prescribed in less than half of those with left ventricular systolic dysfunction. Contraindications to treatment were infrequent and did not explain these findings. PMID- 18402564 TI - Unusual causes of rhabdomyolysis. AB - We report four cases with unusual causes of rhabdomyolysis. These included quail ingestion (coturnism), interaction between grapefruit juice and atorvastatin, hypothyroidism and infection by Epstein-Barr virus. PMID- 18402565 TI - The Wunderly Oration 2007. Indigenous responses to health and disease: principles, goals and pathways. PMID- 18402566 TI - Severe occipital neck pain and a deviating tongue. PMID- 18402567 TI - Mediastinal mass in an asthmatic patient. PMID- 18402568 TI - Tongue infarction in giant cell (temporal) arteritis. PMID- 18402569 TI - Eplerenone in the treatment of Gitelman's syndrome. PMID- 18402570 TI - Growing pains? PMID- 18402572 TI - Conservation, neoliberalism, and social science: a critical reflection on the SCB 2007 Annual Meeting in South Africa. PMID- 18402573 TI - Productivity in conservation research in the southern hemisphere. PMID- 18402574 TI - The popularity of biodiversity. PMID- 18402575 TI - At the end of our tether: the rationality of nonviolence. PMID- 18402576 TI - Nonviolent change, a good idea. PMID- 18402577 TI - Agents of change and nonviolent action. PMID- 18402578 TI - Collapse of bluefin tuna in the Western Atlantic. PMID- 18402579 TI - Conservation performance payments for carnivore conservation in Sweden. PMID- 18402580 TI - The ecological future of the North American bison: conceiving long-term, large scale conservation of wildlife. AB - Many wide-ranging mammal species have experienced significant declines over the last 200 years; restoring these species will require long-term, large-scale recovery efforts. We highlight 5 attributes of a recent range-wide vision-setting exercise for ecological recovery of the North American bison (Bison bison) that are broadly applicable to other species and restoration targets. The result of the exercise, the "Vermejo Statement" on bison restoration, is explicitly (1) large scale, (2) long term, (3) inclusive, (4) fulfilling of different values, and (5) ambitious. It reads, in part, "Over the next century, the ecological recovery of the North American bison will occur when multiple large herds move freely across extensive landscapes within all major habitats of their historic range, interacting in ecologically significant ways with the fullest possible set of other native species, and inspiring, sustaining and connecting human cultures." We refined the vision into a scorecard that illustrates how individual bison herds can contribute to the vision. We also developed a set of maps and analyzed the current and potential future distributions of bison on the basis of expert assessment. Although more than 500,000 bison exist in North America today, we estimated they occupy <1% of their historical range and in no place express the full range of ecological and social values of previous times. By formulating an inclusive, affirmative, and specific vision through consultation with a wide range of stakeholders, we hope to provide a foundation for conservation of bison, and other wide-ranging species, over the next 100 years. PMID- 18402581 TI - Why the conservation of forest genetic resources has not worked. AB - Genetic diversity is indispensable for long-term forest sustainability and is therefore mentioned in numerous binding and nonbinding political covenants calling for action. Nevertheless, there are significant obstacles to the conservation of forest genetic resources. We discuss hindrances to genetic conservation, mainly in Europe. We identified impediments by reviewing the literature and on the basis of the experiences of the authors in this field and their participation in related political processes. The impediments include (1) difficulties in assessing and monitoring genetic erosion and human impacts (e.g., by the lack of markers showing adaptive variation and the lack of record keeping on the use and transfer of forest-tree germplasm), (2) complexities of European national structures that make the development of a common strategy toward forest genetic conservation problematic, (3) lack of effective forest governance in many parts of the world, (4) the general unattractiveness of genes as flagships in raising public awareness, (5) lack of integration of genetic aspects into biodiversity conservation, and (6) the fact that scientists and politicians are often at cross-purposes. To overcome these impediments, forest geneticists and their peers in species conservation have to participate more actively in decision making. In doing so, they must be prepared to face challenges on 2 fronts: participating in political processes and the provision of significant research findings to ensure that decisions with respect to forest genetic diversity are politically implementable and effectively address targets. PMID- 18402582 TI - Assessing the ecological and social benefits of private land conservation in Colorado. AB - Conservation of private land through conservation easements, development agreements, and clustered housing has increased greatly as have criticisms of the laws, public programs, and incentives that motivate landowners to use them. Rapid land-use change at the urban-rural interface in Larimer County, Colorado, has given rise to programs that provide a variety of land-conservation options for landowners. As of January 2005, roughly 60% of Larimer County was publicly owned, and 3% or 16,200 ha was privately owned with some form of protection. We used document analysis, a landowner survey, targeted interviews, and a landscape-level spatial analysis to analyze the patterns, quantities, and qualities of private land conservation. We created a jurisdiction-specific typology of desired benefits from local government-planning documents to help evaluate conservation parcels. Most easements and other conservation documents used general terms and did not describe the site-specific values of the land being conserved. Landowners were able to describe some benefits not included in parcel-specific documents, and our spatial analysis revealed parcel-specific and cumulative conservation benefits such as the amount of buffering, infill, connectivity, protected agricultural land, riparian protection, and other benefits not referenced by either documents or landowners. Conservation benefits provided by a parcel varied depending on its geographic location, the specific institution such as a land trust or open space program that a landowner worked with, and the conservation mechanism used, such as voluntary easement or residential clustering requirements. The methods we used provide a template for jurisdictions wishing to undertake a similar analysis. Our findings may assist other jurisdictions and institutions interested in improving how land-conservation benefits are described; justify and inform future investments in private land conservation; assist local governments and other institutions with the assessment of program effectiveness; and be useful for conservation planners who wish to become more involved in on-the-ground implementation of conservation actions. PMID- 18402583 TI - Cost-effective management alternatives for Snake River Chinook salmon: a biological-economic synthesis. AB - The mandate to increase endangered salmon populations in the Columbia River Basin of North America has created a complex, controversial resource-management issue. We constructed an integrated assessment model as a tool for analyzing biological economic trade-offs in recovery of Snake River spring- and summer-run chinook salmon (Oncorhynchus tshawytscha). We merged 3 frameworks: a salmon-passage model to predict migration and survival of smolts; an age-structured matrix model to predict long-term population growth rates of salmon stocks; and a cost effectiveness analysis to determine a set of least-cost management alternatives for achieving particular population growth rates. We assessed 6 individual salmon management measures and 76 management alternatives composed of one or more measures. To reflect uncertainty, results were derived for different assumptions of effectiveness of smolt transport around dams. Removal of an estuarine predator, the Caspian Tern (Sterna caspia), was cost-effective and generally increased long-term population growth rates regardless of transport effectiveness. Elimination of adult salmon harvest had a similar effect over a range of its cost estimates. The specific management alternatives in the cost effective set depended on assumptions about transport effectiveness. On the basis of recent estimates of smolt transport effectiveness, alternatives that discontinued transportation or breached dams were prevalent in the cost-effective set, whereas alternatives that maximized transportation dominated if transport effectiveness was relatively high. More generally, the analysis eliminated 80-90% of management alternatives from the cost-effective set. Application of our results to salmon management is limited by data availability and model assumptions, but these limitations can help guide research that addresses critical uncertainties and information. Our results thus demonstrate that linking biology and economics through integrated models can provide valuable tools for science-based policy and management. PMID- 18402584 TI - Rates of movement of threatened bird species between IUCN red list categories and toward extinction. AB - In recent centuries bird species have been deteriorating in status and becoming extinct at a rate that may be 2-3 orders of magnitude higher than in prehuman times. We examined extinction rates of bird species designated critically endangered in 1994 and the rate at which species have moved through the IUCN (World Conservation Union) Red List categories of extinction risk globally for the period 1988-2004 and regionally in Australia from 1750 to 2000. For Australia we drew on historical accounts of the extent and condition of species habitats, spread of invasive species, and changes in sighting frequencies. These data sets permitted comparison of observed rates of movement through the IUCN Red List categories with novel predictions based on the IUCN Red List criterion E, which relates to explicit extinction probabilities determined, for example, by population viability analysis. The comparison also tested whether species listed on the basis of other criteria face a similar probability of moving to a higher threat category as those listed under criterion E. For the rate at which species moved from vulnerable to endangered, there was a good match between observations and predictions, both worldwide and in Australia. Nevertheless, species have become extinct at a rate that, although historically high, is 2 (Australia) to 10 (globally) times lower than predicted. Although the extinction probability associated with the critically endangered category may be too high, the shortfall in realized extinctions can also be attributed to the beneficial impact of conservation intervention. These efforts may have reduced the number of global extinctions from 19 to 3 and substantially slowed the extinction trajectory of 33 additional critically endangered species. Our results suggest that current conservation action benefits species on the brink of extinction, but is less targeted at or has less effect on moderately threatened species. PMID- 18402585 TI - Effectiveness of voluntary conservation agreements: case study of endangered whales and commercial whale watching. AB - The use of voluntary approaches to achieve conservation goals is becoming increasingly popular. Nevertheless, few researchers have quantitatively evaluated their efficacy. In 1998 industry, government agencies, and nongovernmental organizations established a voluntary conservation program for whale watching in the northeast region of the United States, with the intent to avoid collisions with and harassment of endangered whales by commercial and recreational whale watching vessels. One important aspect of the program was the establishment of 3 speed zones within specific distances of whales. We wanted to determine the level of compliance with this aspect of the program to gauge its efficacy and gain insights into the effectiveness of voluntary measures as a conservation tool. Inconspicuous observers accompanied 46 commercial whale-watching trips from 12 companies in 2003 (n= 35) and 2004 (n= 11). During each trip, vessel position and speed were collected at 5-second intervals with a GPS receiver. Binoculars with internal laser rangefinders and digital compasses were used to record range and bearing to sighted whales. We mapped whale locations with ArcGIS. We created speed-zone buffers around sighted whales and overlaid them with vessel-track and speed data to evaluate compliance. Speeds in excess of those recommended by the program were considered noncompliant. We judged the magnitude of noncompliance by comparing a vessel's maximum speed within a zone to its maximum recorded trip speed. The level of noncompliance was high (mean 0.78; company range 0.74-0.88), some companies were more compliant than others (p= 0.02), noncompliance was significantly higher in zones farther from whales (p < 0.001), and operators approached the maximum speed capabilities of their vessel in all zones. The voluntary conservation program did not achieve the goal of substantially limiting vessel speed near whales. Our results support the need for conservation programs to have quantifiable metrics and frequent evaluation to ensure efficacy. PMID- 18402586 TI - Economic growth and marine biodiversity: influence of human social structure on decline of marine trophic levels. AB - We assessed the effects of economic growth, urbanization, and human population size on marine biodiversity. We used the mean trophic level (MTL) of marine catch as an indicator of marine biodiversity and conducted cross-national time-series analyses (1960-2003) of 102 nations to investigate human social influences on fish catch and trends in MTL. We constructed path models to examine direct and indirect effects relating to marine catch and MTL. Nations' MTLs declined with increased economic growth, increased urbanization, and increased population size, in part because of associated increased catch. These findings contradict the environmental Kuznets curve hypothesis, which claims that economic modernization will reduce human impact on the environment. To make informed decisions on issues of marine resource management, policy makers, nonprofit entities, and professional societies must recognize the need to include social analyses in overall conservation-research strategies. The challenge is to utilize the socioeconomic and ecological research in the service of a comprehensive marine conservation movement. PMID- 18402587 TI - Dependence of effective marine park zoning on survey design, data quality, and community acceptance: response to Lynch. PMID- 18402589 TI - Threats to avifauna on oceanic islands revisited. PMID- 18402591 TI - Mining and other threats to the New Caledonia biodiversity hotspot. PMID- 18402595 TI - Epidemiological characteristics of tsutsugamushi disease in Oita Prefecture, Japan: yearly and monthly occurrences of its infections and serotypes of its causative agent, Orientia tsutsugamushi, during 1984--2005. AB - Using indirect immunofluorescence assay, we examined the sera of 561 patients from November 1984 to February 2005 to determine the incidence of tsutsugamushi disease (scrub typhus) in Oita Prefecture, Japan. The results obtained were positive in 384 individuals (68.4%). Municipalities where patients were presumed to have been infected with Orientia tsutsugamushi were Taketa City (41.7%), Oyama Town (13.5%), and Ogi Town (8.3%). Infections occurred most often in October, November, and December. A small number of cases occurred from January to May. The serotypes Kuroki (47.5%), Kawasaki (42.5%), and Karp (10.0%) were detected by genetic analysis of O. tsutsugamushi DNA extracted from the blood of 120 patients. The gene sequences of the Kuroki type were highly homologous to that of the Nishino strain. The gene sequences of the Kawasaki type were identical to that of the Kawasaki strain. The gene sequence of the Karp type was highly homologous to that of the JP-2 type. To determine the distribution of vector mites, 558 wild rodents were captured and 72010 mites attached to these rodents were collected from 1982 to 1998. Six genera and 16 species of trombiculid mites were collected. Leptotrombidium pallidum and L. scutellare, which are known to be mite vectors for tsutsugamushi disease, accounted for 20.5% and 5.9%, respectively, of all trombiculid mites collected. The geographical distribution of cases roughly coincided with the distribution of L. scutellare. In Oita Prefecture, L. scutellare is presumed to primarily transmit tsutsugamushi disease. In addition, our results also suggest that L. pallidum transmits the Karp type of the causative rickettsia in some municipalities. PMID- 18402596 TI - Enhancement of gene expression by a peptide p(CHWPR) produced by Bifidobacterium lactis BB-12. AB - Recently, probiotics, including Bifidobacterium, Lactobacillus, and Enterococcus, among other organisms, have been clinically applied for their enhancing effects on defense mechanisms. It is reported that gene expression in somatic cells can be activated by autoinducers, which are hormone-like molecules produced in a microbial QS system. In the present study, based on a hypothesis that a low molecular substance related to the QS system is involved in the probiotics effects of Bifidobacterium, we intended to extract the low-molecular substance. As a result, we successfully isolated the peptide p(CHWPR), which was composed of five amino acids including Cys, His, Trp, Pro, and Arg, and found that the peptide was produced in the stationary phase of bacterial growth and that it could enhance the gene expression of oxalyl-CoA decarboxylase (Oxc). p(CHWPR) enhanced the gene expression of c-myc and interleukin (IL)-6 in an established cell line, HL-60. We demonstrated that p(CHWPR) penetrates the cell membrane and binds specifically to RORgamma, which is a cytosolic nuclear receptor. This suggests that RORgamma bound to p(CHWPR) would bind to promoter regions of the c myc gene. Furthermore, we found that p(CHWPR) also bound to a transcriptional avtivation subunit, CRSP70; this suggests that p(CHWPR), RORgamma, and CRSP70 in combination enhance transcription activity. PMID- 18402597 TI - Efficacy of Clostridium butyricum preparation concomitantly with Helicobacter pylori eradication therapy in relation to changes in the intestinal microbiota. AB - Antibiotic associated diarrhea due to human intestinal microbiota abnormalities is a side effect of H. pylori eradication therapy. We examined intestinal microbiota changes during H. pylori eradication therapy and the preventive effect of CBM588 as a probiotic agent. Nineteen patients with gastro-duodenal ulcer were randomly divided into three groups: group A (without probiotics), group B (with regular doses of CBM588) and group C (with double doses of CBM588). The incidence of diarrhea and soft stools during H. pylori eradication therapy was 43% in group A and 14% in group B, while none of the patients in group C reported diarrhea or soft stools. Both bacterial counts and detection rates of bifidobacteria and/or obligate anaerobe were decreased by eradication therapy. However, bacterial counts of obligate anaerobes in group C were significantly higher than in group A (P < 0.05). Additionally, during eradication therapy C. difficile toxin A was detected in both group A and group B but not in group C. In conclusion, these results indicate that H. pylori eradication therapy induces antibiotic associated diarrhea due to abnormalities in intestinal microbiota and/or C. difficile. However, these side effects might be prevented by probiotics. PMID- 18402598 TI - Identification of a human monoclonal Fab with neutralizing activity against H3N2 influenza A strain from a newly constructed human Fab library. AB - A combinatorial Fab library was constructed in pComb3H phagemid vectors, using RNA from peripheral blood lymphocytes of a healthy volunteer who had recovered from an influenza A virus infection. The library contained approximately 1.3 x 10(8)E. coli transformants. Bio-panning was carried out against an influenza vaccine containing components of influenza A/New Caledonia/20/99 (H1N1), A/Panama/2007/99 (H3N2), and B/Shandong/7/97 for the enrichment of phages displaying human Fab specific to the viral proteins. E. coli transformed with IF1A11, 1 of 94 randomly selected clones, displayed a human Fab antibody molecule (FabIF1A11) with efficient neutralizing activity against H3N2 influenza A virus strains. The purified FabIF1A11 demonstrated neutralizing activity against A/Okayama/6/01 (H3N2) and A/Kitakyushu/159/93 (H3N2) with 50% plaque reduction neutralization titers of 0.11 microg/ml (2.2 nM) and 1.4 microg/ml (28 nM) respectively. However, FabIF1A11 did not show neutralizing activity against the influenza A virus strain A/USSR/77 (H1N1) or the influenza B virus strain B/Kanagawa/73, even at a concentration of 20 microg/ml (400 nM). The Kd of FabIF1A11 was calculated as 3.6 x 10(-9) M. FabIF1A11 was estimated to recognize a conformational epitope on the hemagglutinin of A/Okayama/6/01 (H3N2). The human monoclonal Fab product FabIF1A11 may have potential as a therapeutic or short term prophylactic molecule for humans with influenza A H3N2 infection. PMID- 18402599 TI - Cryptococcus neoformans inhibits nitric oxide synthesis caused by CpG oligodeoxynucleotide-stimulated macrophages in a fashion independent of capsular polysaccharides. AB - Cryptococcus neoformans is eradicated by macrophages via production of NO. Unmethylated CpG-ODN protect mice from infection with this fungal pathogen by inducing IFN-gamma. The present study was designed to elucidate the effect of C. neoformans on the synthesis of NO by alveolar macrophages. For this purpose, MH S, an alveolar macrophage cell line, was stimulated with CpG-ODN in the presence of IFN-gamma. A highly virulent strain of C. neoformans with thick capsule suppressed the production of NO. Capsular polysaccharides were not essential for this suppression, because there was no difference between acapsular mutant (Cap67) and its parent strain. Physical or close interaction of Cap67 with MH-S was necessary, as shown by the loss of such effect when direct contact was interfered by nitrocellulose membrane. Similar effects were observed by disrupted as well as intact Cap67. Whereas the inhibitory effect of intact Cap67 was completely abrogated by heat treatment, disrupted Cap67 did not receive such influence. Finally, disrupted Cap67 did not show any inhibitory effect on the TLR9-mediated activation of NF-kappaB in a luciferase reporter assay with HEK293T cells, although the TLR4-mediated activation was suppressed. These results revealed that C. neoformans suppressed the synthesis of NO by CpG-ODN and IFN gamma-stimulated macrophages in a fashion independent of capsular polysaccharides, although the precise mechanism remains to be elucidated. PMID- 18402600 TI - Evaluation of a rapid ELISA technique for detection of circulating antigens of Toxoplasma gondii. AB - To evaluate a modified rapid ELISA method for detecting CAg during Toxoplasma gondii infection, we analyzed the specificity and sensitivity of the ELISA method by using experimental Toxoplasma infection in rabbits and also tested this method in human samples including 5428 serum, 548 cerebrospinal fluid and two breast milk samples. We prepared PcAb, and used it for rapid one-step sandwich ELISA testing in which an incubation time in the regular ELISA procedure was omitted. This method detected CAg at the concentration of 31.2 ng/mL, and no cross reaction was found with antigens of protozoa (Cryptosporidium parvum, Plasmodium falciparum), trematode (Schistosoma japonicum, Paragonimus sp.) and nematode (Brugia malayi, Ancylostoma duodenale, Ascaris lumbricoides and Trichinella spiralis). CAg was detected in rabbit serum 3 days after infection, and optical density values reached a peak 9-13 days after infection, then declined gradually. Among human serum samples, the positive rate of CAg was 2.11% in cerebral paralysis patients, whereas it was 0.22% or 0.71% in patients without neurological symptoms or in uncomplicated pregnant women. The difference among these three groups was statistically significant (P < 0.05). The positive rate of cerebrospinal fluid samples from cerebral paralysis patients was 10.58%. There is a statistically significant difference between the positive rates of meat-packing workers and blood donors (P < 0.01). In the retrospective analysis, CAg was detected in accordance with the onset of clinical symptoms, suggesting that CAg could reflect the clinical course in humans. Together with these results, CAg detected in the modified rapid sandwich ELISA could be a sensitive marker for acute and active infection of T. gondii. PMID- 18402601 TI - Increased susceptibility to tumor necrosis factor-alpha in butyric acid-induced apoptosis is caused by downregulation of cFLIP expression in Jurkat T cells. AB - Butyric acid is one of the major extracellular metabolites of periodontopathic Gram-negative bacteria. We previously demonstrated that butyric acid induced apoptosis in human T cells. In the present study, we examined the interaction between butyric acid and TNF-alpha in Jurkat T-cell apoptosis. Simultaneous treatment with TNF-alpha enhanced butyric acid-induced apoptosis by promoting caspase activity more than was achieved by either reagent alone. We examined which genes were associated with the increased susceptibility to TNF-alpha caused by butyric acid, and revealed that expression of cFLIP decreased with increased concentrations of butyric acid. Furthermore, exogenous expression of cFLIP protein suppressed the enhancing effect by TNF-alpha in the apoptosis. These results suggest that butyric acid downregulates cFLIP expression and increases the susceptibility to TNF-alpha by activating caspases via the death receptor signal. PMID- 18402602 TI - Salivary gel-forming mucin MUC5B--a nutrient for dental plaque bacteria. AB - INTRODUCTION: Model systems with oral bacteria from dental plaque have demonstrated that the utilization of complex glycoproteins as a food source cannot be undertaken by single species but requires concerted degradation by a multi-species consortium, with each member contributing one or a few hydrolytic enzymes. Unlike previous studies, the aim of the present investigation was to explore the ability of fresh dental plaque to degrade salivary mucin, MUC5B, isolated by methods designed to retain intact the natural polymeric structure and physiological conformation, in an attempt to mimic the naturally occurring interaction between the oral microbiota and salivary mucins. METHODS: Human salivary MUC5B was isolated from whole saliva by density-gradient centrifugation and incubated with freshly isolated supragingival dental plaque with samples subjected to fluorescent staining for viability and metabolic activity. In addition, the degradation of MUC5B oligosaccharide side chains was studied using a lectin assay, recognizing three different carbohydrate epitopes commonly found on mucin oligosaccharide side chains. RESULTS: The addition of purified salivary MUC5B elicited a strong metabolic response from the biofilm cells, whereas individual strains of Streptococcus oralis and Streptococcus gordonii isolated from the same plaque were not able to utilize the MUC5B. The degradation of terminal saccharide moieties on the MUC5B was demonstrated by a marked decrease in both sialic acid and fucose reactivity. CONCLUSION: These results have shown that dental plaque is capable of utilizing human salivary MUC5B as a nutrient source, a process possibly requiring the synergistic degradation of the molecule by a consortium of oral bacteria in the plaque community. PMID- 18402603 TI - Effect of Veillonella parvula on the antimicrobial resistance and gene expression of Streptococcus mutans grown in a dual-species biofilm. AB - INTRODUCTION: Our previous studies showed that Streptococcus mutans and Veillonella parvula dual-species biofilms have a different acid production profile and a higher resistance to chlorhexidine than their single-species counterparts. The aim of the current study was to test whether the susceptibility of S. mutans grown in the presence of V. parvula is also decreased when it is exposed to various other antimicrobials. Furthermore, the aim was to identify other changes in the physiology of S. mutans when V. parvula was present using transcriptomics. METHODS: Susceptibility to antimicrobials was assessed in killing experiments. Transcript levels in S. mutans were measured with the help of S. mutans microarrays. RESULTS: When V. parvula was present, S. mutans showed an increase in survival after exposure to various antimicrobials. Furthermore, this co-existence altered the physiology of S. mutans. The expression of genes coding for proteins involved in amino acid synthesis, the signal recognition particle-translocation pathway, purine metabolism, intracellular polysaccharide synthesis, and protein synthesis all changed. CONCLUSION: Growing in a biofilm together with a non-pathogenic bacterium like V. parvula changes the physiology of S. mutans, and gives this bacterium an advantage in surviving antimicrobial treatment. Thus, the study of pathogens implicated in polymicrobial diseases, such as caries and periodontitis, should be focused more on multispecies biofilms. In addition, the testing of susceptibility to currently used and new antimicrobials should be performed on a multispecies microbial community rather than with single pathogens. PMID- 18402604 TI - Lack of Toll-like receptor 4 decreases lipopolysaccharide-induced bone resorption in C3H/HeJ mice in vivo. AB - INTRODUCTION: Few in vivo studies have demonstrated whether Toll-like receptor 4 (TLR4) is indispensable for lipopolysaccharide (LPS)-induced bone resorption and little is known about the receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) expression induced by LPS under conditions of lack of TLR4. METHODS: We compared bone resorption histomorphometrically in C3H/HeN and C3H/HeJ mice that were repeatedly injected with Actinobacillus actionmycetemcomitans LPS into their gingiva every 48 h. RANKL-, interleukin 1beta- and OPG-positive cells in the connective tissue were also compared immunohistochemically. RESULTS: Bone resorption in C3H/HeJ mice in the fourth, seventh, and tenth injection groups was significantly less than that C3H/HeN mice (P < 0.05). The number of RANKL-positive cells in C3H/HeJ mice in the 10th injection group was significantly smaller than that in C3H/HeN mice (P < 0.05). The numbers of interleukin-1beta-positive cells in C3H/HeJ mice in the seventh and tenth injection groups were significantly decreased compared with those in C3H/HeN mice (P < 0.05). The numbers of OPG-positive cells in C3H/HeN and C3H/HeJ mice gradually increased, but there was no significant difference between the two strains of mice. CONCLUSION: TLR4 is indispensable for LPS-induced bone resorption in vivo. PMID- 18402605 TI - Microbial complexes in supragingival plaque. AB - BACKGROUND/AIMS: To examine microbial communities in supragingival biofilm samples. METHODS: Supragingival plaque samples were taken from 187 subjects at baseline (n = 4745). Fifty-five subjects provided supragingival plaque samples at 1-7 days after professional tooth cleaning (n = 1456); 93 subjects provided 8044 samples between 3 and 24 months post-therapy. All samples were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization. Microbial associations among species were sought using cluster analysis and community ordination techniques for the three groups separately. RESULTS: Six complexes were formed for the baseline samples. Similar complexes were formed for the samples taken 3-24 months post-therapy. However, distinct changes were observed in microbial communities in samples taken during the 7 days of plaque redevelopment. The complexes related to clinical parameters of periodontal disease. CONCLUSION: There were specific microbial complexes in supragingival plaque that were similar to those found in subgingival plaque samples with a few minor differences. The relation of previously unclustered taxa to the complexes was also described. PMID- 18402606 TI - Influences of starch and sucrose on Streptococcus mutans biofilms. AB - INTRODUCTION: The combination of starch and sucrose has been shown to be potentially more cariogenic than either alone. The aim of this study was to examine the influence of starch and sucrose, alone or in combinations, on formation, polysaccharide composition, gene expression, and acidogenicity of Streptococcus mutans biofilms. METHODS: S. mutans UA159 biofilms were formed on saliva-coated hydroxyapatite (sHA) discs in batch culture for 5 days in the presence of 1% (weight/volume) starch, 1% sucrose, 1% starch plus 1% sucrose, 1% starch plus 0.5% fructose plus 0.5% glucose, or 1% sucrose plus 1% glucose. RESULTS: Amylase activity from sHA disks was detected up to 48 h, thereby increasing the availability of reducing sugars and acidogenicity in the early stages of biofilm development. S. mutans grown in the presence of sucrose alone or in combinations formed well-defined and tightly adherent biofilms comprised of mostly water-insoluble polysaccharides (INS); in contrast, the presence of starch or starch + glucose + fructose resulted in little biofilm formation with minimal amounts of INS. However, the combination of starch + sucrose produced biofilms with more biomass and acidogenicity, and a higher content of INS than those grown in sucrose or sucrose + glucose (P < 0.05). The INS extracted from biofilms formed in the presence of starch + sucrose displayed a higher percentage of 3 linked branching (3,4-, 3,6-, and 3,4,6-linked glucose) compared to those from biofilms grown in sucrose or sucrose + glucose. Furthermore, biofilms grown in starch + sucrose expressed significantly higher levels of gtfB messenger RNA than sucrose-grown or sucrose + glucose-grown biofilms (P < 0.05). CONCLUSION: The combination of starch and sucrose has profound effects not only on the composition and structure of the polysaccharide matrix but also on gene expression of S. mutans within biofilms, which may enhance the cariogenic potential of dental biofilms. PMID- 18402607 TI - Role of sucrose in the fitness of Streptococcus mutans. AB - INTRODUCTION: Dental caries has been closely linked to fermentable carbohydrates as key environmental factors. Sucrose has been identified as the most cariogenic carbohydrate. Streptococcus mutans, considered to be the primary pathogen causing dental caries, is able to utilize sucrose as a nutrient source, partially for the production of intracellular storage components and for the production of extracellular glucans via the glucosyltransferases GtfB, GtfC, and GtfD. The following study explores the competitiveness and fitness of S. mutans when grown with different concentrations of sucrose. METHODS: Growth competition with oral streptococci and antimicrobial susceptibility in static biofilm models grown without sucrose or with 0.1% or 0.5% sucrose were investigated using confocal laser scanning microscopy. The numbers of surviving S. mutans of both wild-type and an isogenic Gtf-negative mutant after antimicrobial treatment were determined as colony-forming units. RESULTS: S. mutans was able to establish microcolonies with increasing sucrose concentration in the presence of other streptococcal competitors during biofilm development. The antimicrobial susceptibility decreased when sucrose was available as substrate and was dependent on the presence of the Gtfs. CONCLUSION: The increased resistance against antimicrobial treatment was associated with the availability of sucrose, but was not influenced much by the concentration used during this study. The resistance was strongly associated with the Gtf activity, excluding any intracellular metabolic effect of sucrose in the resistance mechanism. PMID- 18402608 TI - An acute injection of Porphyromonas gingivalis lipopolysaccharide modulates the OPG/RANKL system and interleukin-6 in an ovariectomized mouse model. AB - BACKGROUND/AIMS: In the present study, we attempted to develop a simulated model to explore the causal effects of periodontal pathogens on skeletal homeostasis in postmenopausal osteoporosis. METHODS: Fifty-three female adult ICR mice were randomly assigned to an experimental group (ovariectomized) or a control group. A single injection of Porphyromonas gingivalis lipopolysaccharide (P. gingivalis LPS, ATCC 33277) or Escherichia coli lipopolysaccharide (E. coli-LPS) was administered intraperitoneally 4 weeks after an ovariectomy. Concentrations of interleukin-6 (IL-6), osteoprotegerin (OPG), and the receptor activator of nuclear factor-kappaB ligand (RANKL) in serum were subsequently analyzed using an enzyme-linked immunosorbent assay (ELISA). RESULTS: Under stimulation with P. gingivalis-LPS or E. coli-LPS, the concentration of OPG rose in both groups. The serum level of RANKL showed a decreasing trend 24 h after the injection in both groups. After injection of P. gingivalis-LPS in both the experimental and control animals, the OPG : RANKL ratio increased 24 h after the booster (22.26-620.99, P < 0.05). The serum level of IL-6 in the experimental group significantly increased 1-6 h after administration of E. coli-LPS and 1-3 h after administration of P. gingivalis-LPS (P < 0.05). CONCLUSIONS: A single booster injection of P. gingivalis-LPS induced short-term changes in OPG, RANKL, and IL-6 serum levels in this ovariectomized mouse model. PMID- 18402609 TI - Adherence of Aggregatibacter actinomycetemcomitans via serotype-specific polysaccharide antigens in lipopolysaccharides. AB - INTRODUCTION: Gram-negative Aggregatibacter actinomycetemcomitans is recognized as an important periodontal pathogen. A striking property of this bacterium is its ability to form a tenacious biofilm adhering to abiotic surfaces. Both fimbrial and non-fimbrial adhesins are believed to be responsible for this ability. In our study, specific markerless mutants in the biosynthesis genes of cell surface polysaccharides were constructed with the Cre-loxP recombination system to identify non-fimbrial adhesin(s). METHODS: Non-fimbriated A. actinomycetemcomitans strain ATCC29523 (serotype a) was used to construct a deletion mutant of serotype-a specific polysaccharide antigen (SPA-a) in lipopolysaccharide (LPS). The LPS was purified through a polymyxin B column following phenol extraction, and verified by silver staining following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by immunoblot analysis using rabbit antisera raised against SPA-a. Strains were grown in broth for 2 days and examined for the adherence of bacterial cells on the glass surface. RESULTS: Strain ATCC29523 formed a thin film of bacterial growth on the glass surface. The deletion of SPA-a affected its ability to form this thin film. When this mutant was rescued with the wild-type SPA-a gene cluster, its adherence positive phenotype was restored. CONCLUSION: SPA-a in the LPS molecule appears to promote the adherence of A. actinomycetemcomitans cells to abiotic surfaces. PMID- 18402610 TI - Effect of glucose on Treponema denticola cell behavior. AB - INTRODUCTION: Treponema denticola inhabits the oral subgingival environment and is part of a proteolytic benzoyl-dl-arginine-naphthylamide-positive 'red complex' associated with active periodontal disease. Spirochetes have a unique form of chemotactic motility that may contribute to their virulence. Chemotaxis is essential for efficient nutrient-directed translocation. METHODS: We examined the effect of glucose on T. denticola cell velocity, expression of periplasmic flagella proteins, and chemotaxis, e.g. translocation into capillary tubes. RESULTS: The presence of glucose did not significantly effect T. denticola cell velocity in high viscosity conditions nor did it alter periplasmic flagella protein expression. The addition of glucose to capillary tubes resulted in greater numbers of T. denticola cells in tubes containing glucose. A non-motile mutant did not migrate into capillary tubes containing glucose. CONCLUSION: These results are consistent with a chemotactic response to glucose that is motility dependent. PMID- 18402611 TI - Comparison of nested polymerase chain reaction (PCR), real-time PCR and viral culture for the detection of cytomegalovirus in subgingival samples. AB - INTRODUCTION: The purpose of this study was to compare nested polymerase chain reaction (PCR), real-time PCR, and shell vial for the detection of human cytomegalovirus (HCMV) in subgingival samples in periodontitis patients. METHODS: A group of 44 patients and 24 individuals without periodontitis were included in the study. A full periodontal examination was conducted in each subject. Gingival crevicular fluid (GCF) was collected by pocket lavage and used for viral culture (shell vial). Additional subgingival samples were obtained with paper points and used for molecular analysis. Nested PCR and real-time PCR were used to detect and quantify HCMV. Student's t-test and chi-squared test were used to compare groups. The sensitivity and specificity for the tests were calculated on 2 x 2 tables considering the nested PCR as the gold standard. RESULTS: The detection of HCMV was greater using nested PCR than with either real-time PCR or shell vial (P < 0.0001). However, the frequency detection of both molecular techniques was higher than in viral culture (P < 0.0001). Only one case of chronic periodontitis was positive by viral culture. Agreement between nested PCR and real-time PCR was observed 47.7% and 4.1% of the time in the periodontitis and control groups, respectively. The sensitivity of real-time PCR was 60%, compared with 2.8% for the shell vial technique. CONCLUSIONS: In conclusion, this study confirmed that active HCMV infection occurs in human periodontitis; however, its frequency seems to be low. In contrast, latent periodontal HCMV infection seems to be a more frequent event. PMID- 18402612 TI - Molecular and enzymatic characterization of betaC-S lyase in Streptococcus constellatus. AB - BACKGROUND/AIMS: Streptococcus anginosus and Streptococcus constellatus are frequently isolated from dental abscesses and other suppurative lesions. We previously reported that betaC-S lyase from a strain of S. anginosus produced significantly more hydrogen sulfide than betaC-S lyases from other streptococci. The purpose of this study was to establish the molecular and enzymatic features of the betaC-S lyase in S. constellatus and to elucidate whether this unique capacity is common to many strains of S. constellatus and S. anginosus. METHODS: The capacity of crude extract to produce hydrogen sulfide was evaluated among 16 strains of S. constellatus, S. anginosus, and Streptococcus gordonii. The lcd gene encoding betaC-S lyase was cloned from the genomic DNA of each strain to compare the deduced amino acid sequences. The recombinant betaC-S lyases of three representative strains were purified and characterized. RESULTS: Incubation of crude extracts from all strains of S. constellatus and S. anginosus with l cysteine resulted in the production of a large amount of hydrogen sulfide. The primary sequence of betaC-S lyase was very similar among strains of S. constellatus and S. anginosus. The kinetic properties of the betaC-S lyases purified from S. constellatus resembled those for betaC-S lyases purified from S. anginosus. In contrast, the betaC-S lyases of S. constellatus and S. gordonii differed in terms of their hydrogen sulfide production, with the former producing much more. CONCLUSION: A high level of hydrogen sulfide production, which appears to be a common feature in both S. constellatus and S. anginosus, may be associated with their abscess formation. PMID- 18402613 TI - Purification of bacteriocin LS1 produced by human oral isolate Lactobacillus salivarius BGHO1. AB - INTRODUCTION: Lactobacillus salivarius BGHO1, a human oral isolate with antagonistic activity against growth of Streptococcus mutans, Streptococcus pneumoniae, Staphylococcus aureus, Enterococcus faecalis, Micrococcus flavus, and Salmonella enteritidis, probably produces more than one proteinaceous antimicrobial substance. The objective of this study was the purification of a bacteriocin, named LS1, produced by L. salivarius BGHO1. METHODS: A simple and fast procedure for bacteriocin purification was developed, consisting of reverse phase chromatography of the ammonium sulfate precipitate of cell-free culture supernatant by fast protein liquid chromatography and high-performance liquid chromatography, followed by tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), with the subsequent extraction of bacteriocin from the gel. RESULTS: The supernatant of L. salivarius BGHO1 culture retained its antimicrobial activity after boiling in a water bath for 15 min. Its antimicrobial activity was also maintained even after treatment for 20 min at 121 degrees C in an autoclave. Bacteriocin LS1 was purified to homogeneity. The molecular mass of bacteriocin LS1 was estimated to be approximately 10 kDa, based on tricine SDS-PAGE. During purification, another compound with antimicrobial activity, produced by L. salivarius BGHO1, was detected. The molecular mass of this compound was estimated to be approximately 5 kDa, based on tricine SDS-PAGE. CONCLUSION: Our results imply that LS1 is most probably a new bacteriocin, different from previously described bacteriocins produced by L. salivarius strains. The purification of bacteriocin LS1 enabled the further characterization of LS1 on both the molecular and genetic levels. PMID- 18402614 TI - Actinobacillus actinomycetemcomitans lipopolysaccharide stimulates collagen phagocytosis by human gingival fibroblasts. AB - INTRODUCTION: Collagen phagocytosis by fibroblasts is involved in the intracellular pathway related to collagen breakdown in soft connective tissues. The possible role of lipopolysaccharide (LPS) in regulating this fibroblast function has not been elucidated so we investigated the effect of LPS from Actinobacillus actinomycetemcomitans, a periodontopathic bacterium, on collagen phagocytic activity in human gingival fibroblasts and associated regulatory mechanisms. METHODS: LPS pretreatment stimulated binding of collagen-coated beads to cells and, subsequently, their internalization. RESULTS: The LPS-activated collagen phagocytic process was enhanced in the presence of the soluble form of CD14 (sCD14) or LPS-binding protein (LBP), while the LPS/LBP treatment activated Akt and induced actin reorganization. Furthermore, these LPS/LBP-induced effects were partially suppressed by adding phosphatidyl-inositol-3 kinase (PI3K) inhibitors. CONCLUSION: These results suggest that A. actinomycetemcomitans LPS disturbs the homeostasis of collagen metabolism within gingival tissue by facilitating collagen phagocytosis by gingival fibroblasts, and serum sCD14 and LBP positively regulate the action of LPS. In addition, the PI3K/Akt signaling is thought to partially mediate the LPS/LBP-stimulated collagen phagocytic pathway, which may be dependent on actin cytoskeletal rearrangement. PMID- 18402615 TI - Research and special needs dentistry. PMID- 18402616 TI - Interdisciplinary approach to oral health for persons with developmental disabilities. AB - The aim of this interdisciplinary study was to survey the oral health among persons with developmental disabilities (PDD), and to evaluate the impact of the routine referral process at a regional center in Los Angeles, California. This study evaluated the subjects' oral health, access to care, oral health behaviors, and adherence to referrals. The study recruited 102 subjects (74% were African American or Hispanic). Among them, 24% reported brushing their teeth less than once a day, and 68% had dental insurance. The prevalence of leukoplakia was 4% and the prevalence of temporomandibular joint (TMJ) clicking and pain were 11% and 9%, respectively. Their decayed, missing, filled (DMF) equaled 14 points with 7 missing teeth. The subjects' DMF index was poorer compared to the general population. The routine referral process was not effective in improving the subjects' access to dental care. More sophisticated referral systems need to be developed for this population. PMID- 18402617 TI - Dental findings in Kabuki make-up syndrome: a case report. AB - This article documents the case history of a 9-year-old Brazilian child diagnosed with Kabuki make-up syndrome (KMS). The clinical features are described, with emphasis on the craniofacial and orodental manifestations. The patient had the distinctive KMS craniofacial features consisting of long palpebral fissure, hypertelorism, high-arched eyebrows with sparse hair in the lateral one-third, eyes with eversion of the lateral one-third of the lower eyelids, long and curved eyelashes, palpebral ptosis, large anteverted prominent ears in a lower position, broad, depressed nasal root with a flat nasal tip, and mild neuropsychomotor developmental deficits. The intraoral examination revealed an anterior open bite, overretention of primary teeth, and a high-arched palate. Two findings in this child have not been previously reported in patients with KMS: the presence of supernumerary teeth and taurodontic teeth in the maxillary arch. The clinical and radiographic detection of these unique dental features may be helpful in identifying children who may have milder forms of KMS. PMID- 18402618 TI - Death, dying and bereavement: a survey of dental practitioners. AB - The dentist's role following the death of a patient in his/her practice has received little attention in the literature. This study determined the prevalence of death within a dental practice. It also investigated methods by which dentists supported grieving survivors, and how frequently dentists received formal bereavement education in dental school. A perceived need for training in death and dying was also investigated. A survey was mailed to 200 randomly selected general dental practitioners in Ontario, Canada. It was found that (1) the vast majority of respondents (86%) had experienced the death of a patient within the past 12 months; (2) support methods included sending sympathy cards (79.3%), sending flowers (34.5%), attending the funeral or wake (23%), or visiting/calling survivors (11.5%); (3) only 4% of respondents reported receiving formal bereavement education during dental school; and (4) 61% believed bereavement education should be included in the dental school curricula. While the majority of dentists in this study provided bereavement support and believed they could effectively comfort grieving persons, these dentists experienced significant stress when dealing with issues of death and bereavement. The stress may be explained by inadequate education and exposure to the issues of death and dying. PMID- 18402619 TI - Prevalence of dental trauma in patients with cerebral palsy. AB - This study presents the findings of a record review that evaluated the prevalence of dental trauma in patients with cerebral palsy and evaluated its possible relationship with age, type of palsy, and epilepsy. The dental records of 2,200 patients with special needs admitted to the special care clinic of the School of Dentistry of Aracatuba/UNESP, Brazil, between 1998 and 2003 were reviewed. Of the records that were analyzed, 500 patients who had cerebral palsy were selected for this study. Information regarding age, gender, type of palsy and dental trauma was collected and statistically analyzed. It was observed that 10.6% of the subjects (n = 53) had sustained dental trauma. The number of traumatized teeth was 84. Enamel or enamel/dentin fractures were the most frequent types of traumatic injury (84.9%). The permanent maxillary central incisors were the most commonly affected teeth (50%). The frequency of traumatic injuries showed no significant correlation (p> .05) with the type of cerebral palsy or gender and they were more frequent in subjects between the ages of 0 and 12 years. Having epilepsy was not a statistically significant risk factor (p> .05) for dental trauma. PMID- 18402620 TI - Oral health, dental treatment, and cardiac valve surgery outcomes. AB - The aim of this study was to determine whether not treating chronic dental infection during the admission for cardiac valve surgery would increase the morbidity and mortality of patients. Patients were divided into three groups: dentally unhealthy and untreated (Group A), dentally healthy not requiring treatment (Group B), and dentally unhealthy and treated (Group C). Hospital computer records and phone interviews were used to assess morbidity and mortality as assessed through the Social Security Death Index. Ninety-eight patient charts were reviewed. Patients in Group A (n = 47)were not at a significantly greater risk for developing infective endocarditis (IE) within 6 months of cardiac surgery compared to patients in Groups B (n = 17) and C (n = 34). Also, patients in Group A did not have a significantly higher rate of mortality compared to other groups (p= .09). The results suggest that there is no need to treat chronic oral infections in patients with compromised cardiac function within 24 to 48 hours prior to cardiac valve replacement surgery since this will not lower the risk of IE and death following cardiac valve surgery. Multicenter prospective case-controlled studies are needed to address this question definitively. PMID- 18402621 TI - Doxepin rinse for management of mucositis pain in patients with cancer: one week follow-up of topical therapy. AB - This study assessed the effectiveness of oral doxepin rinse for mucositis-related pain management in patients following 1 week of repeated dosing. Patients with oral mucositis due to head and neck radiation therapy or hematopoietic stem cell transplant (HSCT) were recruited to participate in a 1-week follow-up study. Subjects who gave informed consent rinsed with doxepin (5 ml) during the initial visit and were then told to use doxepin rinse over the next week as needed, three to six times per day, and return for a follow-up visit. At each visit, mucositis was scored using the Oral Mucositis Assessment Scale and oral pain was assessed using a visual analogue scale before and after rinsing. The use of a systemic analgesic was recorded, and side effects were documented. At the follow-up visit, subjects were also asked to retrospectively report average pain scores they experienced over the past week, 5 and 15 minutes following rinse. Nine subjects were enrolled in the study. Statistically significant reductions in pain scores were reported for 2 hours following doxepin rinse during the initial visit (p < .05). Patients recalled that their pain significantly dropped within 5 minutes of rinsing over the week of repeated dosing (p < .05). At the follow-up visit, subjects reported statistically significant pain reduction 5 minutes after doxepin rinsing (p < .05). These results indicate that doxepin rinsing continues to produce reduced intensity of pain levels over a 1-week span of repeated dosing. PMID- 18402623 TI - Norepinephrine and serotonin imbalance in the locus coeruleus in bipolar disorder. AB - OBJECTIVES: Abnormalities in norepinephrine (NE) and serotonin (5-HT) are implicated in bipolar disorder (BD). We examined 5-HT input and NE neurons in the locus coeruleus (LC, the NE nucleus that innervates the forebrain) in BD by quantifying immunoreactivity (IR) for tyrosine hydroxylase (TH) and tryptophan hydroxylase (TPH), the biosynthetic enzymes for NE and 5-HT, respectively. METHODS: Six suicides with BD were compared to matched normal controls and unipolar major depression suicides, using immunocytochemistry with computer assisted quantification of immunoreactivity. RESULTS: Depressed bipolar suicides had 26.7 +/- 1.3% of LC area occupied by the TH immunoreactive (TH-IR) process, while controls had 50.7 +/- 8% (p = 0.002) and unipolar depressed suicides had 50.3 +/- 2.5% (p = 0.003). In bipolars, these processes did not stain as darkly (1.9 +/- 0.5 x background) as controls (2.9 +/- 0.9 x background; p = 0.01) or unipolars (2.9 +/- 0.6 x background; p = 0.002). Bipolar suicides also had less TPH-IR processes in the LC (11.7 +/- 10%) compared with controls (32.8 +/- 8.8%; p = 0.01) or unipolar suicides (30.3 +/- 8%; p = 0.02). The TPH-IR intensity did not differ between groups. CONCLUSIONS: We found less TH-IR and TPH-IR in the LC in depressed bipolar suicides, but not unipolar suicides, suggesting that both NE and 5-HT activity is lower in BD. Studies during manic or euthymic states will determine whether these changes are mood state dependent. PMID- 18402624 TI - Neuroprotective effect of chronic lithium treatment against hypoxia in specific brain regions with upregulation of cAMP response element binding protein and brain-derived neurotrophic factor but not nerve growth factor: comparison with acute lithium treatment. AB - OBJECTIVES: We evaluated the neuroprotective effect of chronically or acutely administered lithium against hypoxia in several brain regions. Furthermore, we investigated the contribution of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), and cAMP response element binding protein (CREB) to the neuroprotective effect of lithium. METHODS: Brain slices were prepared from rats that had been treated chronically or acutely with lithium. The cerebral glucose metabolic rate (CMRglc) before and after hypoxia loading to brain slices was measured using the dynamic positron autoradiography technique with [(18)F]2 fluoro-2-deoxy-D-glucose. The changes of expression of proteins were investigated using Western blot analysis. RESULTS: Before hypoxia loading, the CMRglc did not differ between the lithium-treated and untreated groups. After hypoxia loading, the CMRglc of the untreated group was significantly lower than that before hypoxia loading. However, the CMRglc of the chronic lithium treatment group recovered in the frontal cortex, caudate putamen, hippocampus and cerebellum, but not in the thalamus. In contrast, the CMRglc of the acute lithium treatment group did not recover in any analyzed brain regions. After chronic lithium treatment, the levels of expression of BDNF and phospho-CREB were higher than those of untreated rats in the frontal cortex, but not in the thalamus. However, the expression of NGF did not change in the frontal cortex and thalamus. CONCLUSIONS: These results demonstrated that lithium was neuroprotective against hypoxia only after chronic treatment and only in specific brain regions, and that CREB and BDNF might contribute to this effect. PMID- 18402625 TI - A new accurate method for predicting lithium clearance and daily dosage requirements in adult psychiatric patients. AB - OBJECTIVE: The present study aimed to derive new equations for estimating lithium clearance and daily dosage requirements needed to achieve an intended lithium serum level for adult psychiatric inpatients and outpatients. METHODS: Data were retrospectively collected from 60 adult psychiatric patients (34 males and 26 females, aged between 18-80 years) in both inpatient and outpatient settings. All variables that might affect lithium clearance and/or lithium serum concentration were included and analyzed by stepwise multiple linear regression to produce equations describing lithium clearance and daily dosage requirements for these patients. The validation of the developed equations was performed by application to another 60 psychiatric subjects in both the inpatient and outpatient settings. The bias and accuracy of the new methods were also compared to those set forth by the empirical method and the a priori methods developed by Zetin, Pepin, Jermain and Terao and colleagues. RESULTS: The following prediction equations for lithium clearance (CL(Li)) were obtained: CL(Li) (inpatients) = 0.932 + 0.185CL(Cr) and CL(Li) (outpatients) = 1.021 + 0.141CL(Cr). The equations derived for daily dosage requirements were: daily dose (inpatients, mg) = 350.15 + 289.92 (desired lithium level, mmol/L) + 0.84 (weight, kg) - 1.76 (age, years) + 34.43 [tricyclic antidepressant (TCA), yes = 1, no = 0] + 62.1(CL(Cr), L/h) + 13.1 [blood urea nitrogen (BUN), mmol/L] + 40.9 (sex, male = 1, female = 0) and daily dose (outpatients, mg) = 784.92 + 530.22 (desired lithium level, mmol/L) + 8.61 (weight, kg) - 12.09 (age, years) - 11.14 (TCA, yes = 1, no = 0) - 7.63 (CL(Cr), L/h) - 42.62 (BUN, mmol/L) - 23.43 (sex, male = 1, female = 0). In the present method, the prediction error for clearance was 10.31% and 6.62% for inpatients and outpatients, respectively, and the prediction error for daily dosage requirements was 3.96% and 2.95% for inpatients and outpatients, respectively. CONCLUSIONS: Compared to previously reported methods, the present method proved to be accurate and can be safely used for the prediction of lithium clearance and daily dosage requirements in psychiatric inpatients and outpatients. PMID- 18402626 TI - Auditory P300 in patients with bipolar disorder and their unaffected relatives. AB - OBJECTIVES: There is evidence that genetic susceptibility may be shared between bipolar disorder (BD) and schizophrenia, but electrophysiological phenotypes which have been extensively used in studies of genetic susceptibility for schizophrenia remain far less explored in bipolar illness. This study assesses whether auditory P300 latency delays and amplitude reductions, which have been demonstrated in patients with schizophrenia and their unaffected first-degree relatives, are associated with familial liability to psychotic bipolar illness. METHODS: The P300 auditory evoked potential was obtained using an oddball task from 37 participants with BD who had a history of psychotic symptoms, 38 of their unaffected first-degree relatives and 42 healthy unrelated comparison subjects. Patients and relatives came from families multiply affected with BD or another functional psychotic disorder. P300 amplitude and latency at midline sites were compared between the groups, using linear regression analyses and robust variance estimators for clustered data, including age and gender as covariates. RESULTS: Bipolar disorder patients with a history of psychosis and their unaffected relatives showed significantly delayed P300 latency at Pz compared to controls. The groups did not differ in P300 amplitude. CONCLUSIONS: P300 latency delays are associated with both psychotic BD and familial liability for this illness. Sample size limited our ability to test for multimodal distribution of P300 measures among relatives, which might be expected if only a subgroup inherits any deficits. In future it will be of interest to directly compare groups of families with psychotic and non-psychotic forms of BD to explore further the role of psychotic symptoms with regard to P300 measures in the disorder. Our results indicate that delayed P300 latency is a promising candidate endophenotype for psychotic BD, as well as schizophrenia, and may reflect the impact of shared susceptibility genes for both types of psychosis. PMID- 18402627 TI - Neurocognitive endophenotypes for bipolar disorder. AB - OBJECTIVES: Neurocognitive deficits have been proposed as vulnerability markers or endophenotypes for the development of bipolar I disorder (BD I). However, few research studies have examined whether neurocognitive deficits also exist in first-degree relatives of individuals with BD I. METHODS: This prospective study examined neurocognitive function in individuals with BD I, their first-degree relatives and a normal control group using a comprehensive battery of neurocognitive tests. RESULTS: Results indicated that individuals with bipolar disorder and their unaffected relatives demonstrated neuropsychological deficits in comparison to the normal control group in the domains of visuospatial/constructional abilities, executive function, visual learning and memory, and motor speed. In general, the unaffected relatives demonstrated an intermediate level of performance in comparison to the normal control and bipolar group. After adjustment for mood symptoms, significant differences were present for the visuospatial/constructional, executive function, and motor domains. Individuals with bipolar disorder also demonstrated a differential right versus left hemisphere deficit with respect to neurocognitive tasks. CONCLUSIONS: Results suggest that deficits on specific neuropsychological tests, most notably Digit Symbol, Block Design and Judgment of Line Orientation, may be indicative of cognitive endophenotypes for bipolar disorder. Replication studies are needed to further identify these deficits as endophenotypes for BD I. PMID- 18402628 TI - Adaptation and validation of the Spanish version of the Mood Disorder Questionnaire for the detection of bipolar disorder. AB - OBJECTIVE: The Mood Disorder Questionnaire (MDQ) is an instrument for the detection of patients with bipolar disorder (BD). The original English version is validated in both the psychiatric and the general population, but a validated Spanish version is not yet available. Psychometric properties of the Spanish adaptation of the MDQ in psychiatry are described. METHODS: The MDQ is a self administered questionnaire comprising a list of 13 hypomanic symptoms and two questions about concurrence of symptoms and functional impairment caused by the symptoms. We selected patients from 15 psychiatric outpatient departments, diagnosed with BD type I and II (BDI and BDII) and major depression (MD) according to DSM-IV-TR criteria (concurrent validity instrument). A control group of healthy subjects (HS) was selected. The patient-selection criteria included stability of the disorder and pharmacological treatment. The MDQ was administered to 236 subjects, distributed among the four groups, on two occasions, four weeks apart. We analysed the internal consistency, test-retest reliability, and discriminative capacity of the MDQ for the detection of patients with BD. RESULTS: Concurrent validity based on diagnosis according to DSM-IV-TR was 0.83. The internal consistency, evaluated by Cronbach's alpha, was 0.90. The mean (SD) number of affirmative responses by group was: 9.8 (2.4) for BDI, 8.5 (2.8) for BDII, 2.7 (2.2) for MD, and 1.02 (1.9) for HS. Statistically significant differences between all the groups were found (Kruskal-Wallis test, p < 0.001). Concurrent validity using the diagnostic variable was 0.83. Test-retest reliability was 0.92. We analysed the scale's discriminative capacity, revealing a sensitivity value of 0.60 [95% confidence interval (CI) = 0.51-0.69] and a specificity value of 0.98 (95% CI = 0.94-0.99) in the detection of BD. The positive and negative probability ratios were 35.5 and 2.4, respectively. If we consider only seven positive responses as the discriminative criterion, sensitivity increases to 0.81 (95% CI = 0.73-0.88), the specificity value is 0.95 (95% CI = 0.89-0.98) and the positive and negative probability quotients are 16 and 5.3. CONCLUSIONS: The psychometric characteristics of the Spanish version are similar to those of the original version. In the Spanish adaptation of the MDQ, seven positive responses to hypomanic symptoms show a good discriminative capacity for BD in patients attending psychiatric outpatient facilities; therefore, this cut-off score is proposed for the detection of BD in psychiatric outpatients. PMID- 18402629 TI - Differences in outcome of DSM-IV bipolar I and II disorders. AB - OBJECTIVES: To investigate whether the course of bipolar disorder (BD) type II is more depressive than that of BD I, and, if so, to explore the underlying factors that cause this difference. METHODS: In a prospective, naturalistic study of 191 secondary care psychiatric in- and outpatients diagnosed in an acute phase of BD I or II, 160 patients (85.1%) were followed for 18 months. Using a life chart, the exact timing of symptom states in follow-up was examined. Differences between BD I (n = 75) and II (n = 85) in duration of index phase and episode, time to full remission and recurrence, and time in any mood episode were investigated. RESULTS: Patients with BD II spent a higher proportion of time ill (47.5% versus 37.7%; p = 0.02) and in depressive symptom states (58.0% versus 41.7%; p = 0.003) than BD I patients. This was a result of the higher proportion (61.7% versus 48.6%; p = 0.03) and mean number (1.69 versus 1.11; p = 0.006) of depressive illness phases in BD II, rather than of differences in the duration of depressive phases. Type of index phase strongly predicted the outcome. In linear regression models, both BD II and type of index phase predicted more time spent in depressive symptom states. CONCLUSIONS: In medium-term follow-up, BD II patients spend about 40% more time in depressive symptom states than BD I patients because a higher proportion of BD II patients have depressive phases and the frequency of these is higher. Differences in type of index phase may markedly confound differences in outcome between BD I and II. PMID- 18402630 TI - A preliminary functional magnetic resonance imaging study of prefrontal-amygdalar activation changes in adolescents with bipolar depression treated with lamotrigine. AB - OBJECTIVES: Hypotheses regarding mood dysregulation in bipolar disorder (BD) have centered on limbic overactivity with relative prefrontal underactivity during mood episodes. Therefore, we hypothesized that adolescents with bipolar depression successfully treated with lamotrigine would show decreases in amygdalar activation, and increases in prefrontal activation. METHODS: Eight adolescents with BD underwent functional magnetic resonance imaging (fMRI) at baseline and after eight weeks of lamotrigine treatment. Blocks of negatively and neutrally valenced emotional pictures were presented during scanning, and subjects were asked to rate how each picture made them feel. Activation in bilateral amygdalae and dorsolateral prefrontal cortices (DLPFC) for negative minus neutral pictures was correlated with Children's Depression Rating Scale (CDRS) scores. RESULTS: Mean (SD) CDRS scores decreased significantly, from 53.0 (10.6) at baseline to 26.3 (5.3) at Week 8. This clinical improvement was correlated with decreased right amygdalar activation (r = 0.91, p = 0.002). At Week 8, but not baseline, CDRS score was positively correlated with bilateral amygdalar activation (r = 0.85, p = 0.007). DLPFC activation was not correlated with change in CDRS score. CONCLUSIONS: These preliminary results indicate that adolescents with BD treated with lamotrigine demonstrated less amygdalar activation when viewing negative stimuli as depressive symptoms improved. Larger controlled studies are needed to confirm these findings. PMID- 18402631 TI - Lamotrigine in bipolar disorder: efficacy during pregnancy. AB - OBJECTIVE: Clinical management of bipolar disorder (BPD) patients during pregnancy is a major challenge. The high risk of bipolar depression during pregnancy encourages consideration of lamotrigine (LTG). We therefore compared recurrence risks among pregnant women with BPD treated with LTG to those discontinuing mood stabilizer therapies. METHODS: We compared risks and weeks to new DSM-IV illness-episodes among 26 initially clinically stable pregnant women diagnosed with DSM-IV BPD who continued LTG treatment to those discontinuing all mood stabilizer treatment during pregnancy. RESULTS: The risk of new illness episodes with LTG was 30% versus 100% after discontinuing mood stabilizers, and survival-computed time-to-25%-recurrence was 28.0 versus 2.0 weeks (chi(2 )=17.3, p < 0.0001; hazard ratio = 12.1; 95% confidence interval = 1.6-91.7). CONCLUSIONS: Discontinuing mood stabilizer treatment presents high risks of illness-recurrence among pregnant women diagnosed with BPD. LTG may afford protective effects in pregnancy, and its reported fetal safety compares favorably to other agents used to manage BPD. PMID- 18402632 TI - Functional impairment as a predictor of short-term symptom course in bipolar I disorder. AB - OBJECTIVES: Most prior research has focused on functional impairment as a consequence, rather than a predictor, of mood symptoms in bipolar disorder (BD). Yet the majority of this research has been cross-sectional, thus limiting conclusions regarding directionality of effects. Indeed, just as functional impairment may represent an important outcome of BD, it may also serve as a risk factor for future affective symptoms or episodes. Thus, the primary aim of this study was to evaluate functional impairment as a predictor of mood symptoms in BD. METHODS: Ninety-two patients with bipolar I disorder, recruited from hospital settings, were administered the Modified Hamilton Rating Scale for Depression, Bech-Rafaelson Mania Scale, and UCLA Social Attainment Survey (SAS) at baseline and at four-month follow-up. RESULTS: Overall, patients evidenced a moderate level of functional impairment at both time points. Whereas baseline functional impairment was not associated with subsequent manic symptoms, baseline functional impairment was significantly predictive of depressive symptom levels at four month follow-up. When individual SAS subscales were evaluated, impaired romantic relationship functioning and activity involvement were each significantly predictive of subsequent depressive symptoms, whereas baseline peer functioning was not. CONCLUSIONS: The study results suggest that functional impairment may be predictive of subsequent depressive, but not manic, symptoms over a relatively short-term follow-up period. Future studies that evaluate illness course over longer follow-up periods would be useful to further clarify the potential bidirectional relationship between depression and functional impairment in BD. PMID- 18402633 TI - Age at onset in Sardinian bipolar I patients: evidence for three subgroups. AB - OBJECTIVE: We studied age at onset (AAO) in order to assess the presence of different subgroups in a homogeneous genetic population, such as the Sardinian population. METHODS: Admixture analysis was applied in order to identify a model of separate normal distribution of AAO characterized by different means, variances and population proportions to allow for evaluation of different subgroups in a sample of 181 unrelated patients of Sardinian origin with bipolar disorder (BP) type I. The Mann-Whitney test was used to compare the means of AAO between subjects with a history of suicide attempts and subjects with no such history. RESULTS: The best-fitting model had three components with means (SD) of 18.1 (2.3), 24.3 (5.3) and 41 (11.5) years, comprising 36%, 39% and 25% of the sample, respectively. We obtained two cut-off points at 21 and 33 years, enabling the sample to be divided into three subgroups. The Mann-Whitney test revealed a difference between the mean AAO of subjects with a positive history of suicide attempts and that of subjects with no such history (p = 0.041). CONCLUSIONS: We found three AAO sub-groups in our sample of BP I patients of Sardinian origin. Our findings add further support to the hypothesis whereby AAO acts as a clinical marker of biological heterogeneity in BP. PMID- 18402634 TI - Mania as the first manifestation of Wilson's disease. AB - BACKGROUND: Although mental changes are frequent in Wilson's disease, severe psychiatric disorders occur uncommonly and usually accompany the neurological picture. There are few reports in the literature of Wilson's disease patients with typical bipolar affective disorder (BPAD). CASE REPORT: The authors report the case of a patient with Wilson's disease whose initial manifestation was a manic episode followed by depression. Tremor in the upper limbs appeared one year after the onset of symptoms. The diagnosis of Wilson's disease was established three years after the first symptoms appeared, based on the neuropsychiatric picture, the detection of Kayser-Fleischer rings and the results of diagnostic tests indicating chronic liver disease and copper excess. ATP7B genotyping and magnetic resonance imaging of the brain with proton spectroscopy study were also performed. The patient became asymptomatic two years after starting treatment with penicillamine and remained non-symptomatic controlled during the eight-year follow-up period, without any specific treatment for the BPAD. CONCLUSIONS: To our knowledge, this is a singular report of a case of Wilson's disease in which a manic episode preceded the onset of neurological symptoms. The association between Wilson's disease and bipolar disorder is discussed. PMID- 18402636 TI - New perspectives in neurogastroenterology and motility. Foreword. PMID- 18402637 TI - A 20-year perspective--from Journal of Gastrointestinal Motility to Neurogastroenterology and Motility. AB - This review traces the origins of this journal as the consequence of a revolution in academic publishing that started in Oxford 50 years ago. It also traces the expansion of the field of gastrointestinal motility by the development of new methodology that prompted both the creation of learned societies and a role for a journal devoted to this field. The problems that arose because the new journal was an 'orphan journal' lacking a parent society are recounted, as are the events that eventually led to its adoption. Finally, the introduction of the term 'neurogastroenterology' into scientific and clinical parlance is explained. PMID- 18402638 TI - The journey to establish purinergic signalling in the gut. AB - Although the concept of purinergic signalling arose from experiments designed to find the identity of the non-adrenergic, non-cholinergic (NANC) inhibitory neurotransmitter in the gut, it has taken many years for the more general importance of the various roles of ATP as a physiological messenger in the gut to be recognized. Firstly, vasoactive intestitial polypeptide (VIP) and later nitric oxide (NO) were considered the NANC transmitter and it was only later, after the concept of cotransmission was established, that ATP, NO and VIP were recognized as cotransmitters in NANC nerves, although the proportions vary in different gut regions. Recently, many purinoceptor subtypes have been identified on myenteric, submucosal motor, sensory and interneurons involved in synaptic neurotransmission and neuromodulation and reflex activity of several kinds, including ascending excitatory and descending inhibitory reflex pathways. Nucleotide receptors have been shown to be expressed on enteric glial cells and interstitial cells of Cajal. Purinergic mechanosensory transduction, involving release of ATP from mucosal epithelial cells during distension to stimulate subepithelial nerve endings of intrinsic and extrinsic sensory nerves to modulate peristalsis and initiate nociception respectively, is attracting current attention. Exciting new areas of interest about purinergic signalling in the gut include: involvement of purines in development, ageing and regeneration, including the role of stem cells; studies of the involvement of nucleotides in the activity of the gut of invertebrates and lower vertebrates; and the pathophysiology of enteric purinergic signalling in diseases including irritable bowel syndrome, postoperative ileus, oesophageal reflux, constipation, diarrhoea, diabetes, Chaga's and Hirschprung's disease. PMID- 18402639 TI - Functional development of the enteric nervous system--from migration to motility. AB - The enteric nervous system (ENS) consists of many different types of enteric neurones forming complex reflex circuits that underlie or regulate many gut functions. Studies of humans with Hirschsprung's disease (distal aganglionosis), and of animal models of Hirschsprung's disease, have led to the identification of many of the genetic, molecular and cellular mechanisms responsible for the colonization of the gut by enteric neurone precursors. However, later events in the ENS development are still poorly understood, including the development of functioning ENS circuits. This article is a personal view of the current state of play in our understanding of the ENS development and of the future of the field. PMID- 18402640 TI - The enteric nervous system: normal functions and enteric neuropathies. AB - Most aspects of the normal organisation and functioning of the enteric nervous system have been resolved in recent years, especially for the small and large intestines, where the ENS has essential roles in controlling bowel movement and transmucosal fluid exchange. The roles of the ENS in the esophagus are not understood, and the relative roles of intrinsic reflexes in relation to extrinsic control of the stomach require clarification. In the small intestine and colon, it needs to be understood how neural activity is orchestrated to subserve different functional outcomes, for example propulsion, mixing and retrograde movement. However, the most important future challenges are to properly understand the molecular and cellular changes that underlie enteric neuropathies, to utilise knowledge of the normal neurochemistry, pharmacology and physiology of the ENS to devise strategies to treat disorders of motility and secretion, and to develop effective therapeutic compounds. It is suggested that ion channels of enteric neurons have been under-investigated as therapeutic targets. Other future challenges lie in the identification of biomarkers for functional bowel disorders and in the use of neural stem cells for restitution of ENS function. PMID- 18402641 TI - Regulation of smooth muscle excitation and contraction. AB - Smooth muscle cells (SMC) make up the muscular portion of the gastrointestinal (GI) tract from the distal oesophagus to the internal anal sphincter. Coordinated contractions of these cells produce the motor patterns of GI motility. Considerable progress was made during the last 20 years to understand the basic mechanisms controlling excitation-contraction (E-C) coupling. The smooth muscle motor is now understood in great molecular detail, and much has been learned about the mechanisms that deliver and recover Ca2+ during contractions. The majority of Ca2+ that initiates contractions comes from the external solution and is supplied by voltage-dependent Ca2+ channels (VDCC). VDCC are regulated largely by the effects of K+ and non-selective cation conductances (NSCC) on cell membrane potential and excitability. Ca2+ entry is supplemented by release of Ca2+ from IP(3) receptor-operated stores and by mechanisms that alter the sensitivity of the contractile apparatus to changes in cytoplasmic Ca2+. Molecular studies of the regulation of smooth muscle have been complicated by the plasticity of SMC and difficulties in culturing these cells without dramatic phenotypic changes. Major questions remain to be resolved regarding the details of E-C coupling in human GI smooth muscles. New discoveries regarding molecular expression that give GI smooth muscle their unique properties, the phenotypic changes that occur in SMC in GI motor disorders, tissue engineering approaches to repair or replace defective muscular regions, and molecular manipulations of GI smooth muscles in animals models and in cell culture will be topics for exciting investigations in the future. PMID- 18402642 TI - Interstitial cells of Cajal in health and disease. AB - The gastrointestinal tract serves the physiological function of digesting and absorbing nutrients from food and physically mixing and propelling these contents in an oral to anal direction. These functions require the coordinated interaction of several cell types, including enteric nerves, immune cells and smooth muscle. Interstitial cells of Cajal (ICC) are now recognized as another cell type that are required for the normal functioning of the gastrointestinal tract. Abnormalities in ICC numbers and networks are associated with several gastrointestinal motility disorders. This review will describe what is known about the function and role of ICC both in health and in a variety of motility disorders with a focus on unresolved issues pertaining to their role in the control of gastrointestinal motility. PMID- 18402644 TI - Postinflammatory visceral sensitivity and pain mechanisms. AB - The inflammatory reaction is normally tightly regulated, and as soon as the original insult has been cleared, a resolution phase starts that aims at leading the tissues back to a normal physiological state. However, after intestinal inflammation, a number of patients develop postinflammatory hypersensitivity symptoms, which can be defined as an excessive sensitivity to gut nociceptive stimulation. The pain experienced by those patients has been largely studied in the context of postinfectious intestinal diseases. The mechanisms of postinflammatory persistent visceral pain involve peripheral and central neuroplastic changes, low-grade chronic inflammation that sensitizes visceral afferent pathways and sensitization of non-neuronal resident cells of the gut. Several molecular determinants such as neurokinins, serotonin, proteases and voltage-gated ion channels seem to play a significant role in the control of postinflammatory visceral sensation. This review tries to give insights into the mechanisms of persistent visceral pain following the resolution of intestinal inflammation and tries to identify what needs to be done to further advance the field of postinflammatory hypersensitivity clinical management. PMID- 18402643 TI - Vagal and hormonal gut-brain communication: from satiation to satisfaction. AB - Studying communication between the gut and the brain is as relevant and exciting as it has been since Pavlov's discoveries a century ago. Although the efferent limb of this communication has witnessed significant advances, it is the afferent, or sensory, limb that has recently made for exciting news. It is now clear that signals from the gut are crucial for the control of appetite and the regulation of energy balance, glucose homeostasis, and more. Ghrelin, discovered just a few years ago, is the first gut hormone that increases appetite, and it may be involved in eating disorders. The stable analogue of glucagon-like peptide 1 has rapidly advanced to one of the most promising treatment options for type-2 diabetes. Changes in the signalling patterns of these and other gut hormones best explain the remarkable capacity of gastric bypass surgery to lower food intake and excess body weight. Given the enormous societal implications of the obesity epidemic, these are no small feats. Together with the older gut hormone cholecystokinin and abundant vagal mechanosensors, the gut continuously sends information to the brain regarding the quality and quantity of ingested nutrients, not only important for satiation and meal termination, but also for the appetitive phase of ingestive behaviour and the patterning of meals within given environmental constraints. By acting not only on brainstem and hypothalamus, this stream of sensory information from the gut to the brain is in a position to generate a feeling of satisfaction and happiness as observed after a satiating meal and exploited in vagal afferent stimulation for depression. PMID- 18402645 TI - Mentation on the immunological modulation of gastrointestinal motility. AB - The immunological modulation of gastrointestinal motility is currently one of the most dynamic and fascinating areas of enteric research, as investigators are beginning to focus their studies on the pathophysiology of various gastrointestinal dysmotilities. The new fruits of this investigative initiative has resulted in the appearance of a fascinating series of articles which demonstrate that intestinal inflammatory events alter a distinct population of enteric neurons and that these alterations last long past the apparent resolution of the inciting event. Studies over the past few years have unequivocally demonstrated that the muscularis externa itself is an active and complex immunological compartment with unique features. The rodent muscularis externa is constitutively populated by a dense network of muscularis macrophages throughout the entire gastrointestinal tract. Although few other leukocytes are present in the rodent, the human muscularis is densely populated by both macrophages and mast cells. Postoperative ileus and endotoxin-induced ileus have turned out to be extremely useful rodent models to elucidate the importance of muscularis leukocytes in causing intestinal dysfunction. Using models of ileus, studies have demonstrated that a complex molecular inflammatory scenario is triggered within the muscularis externa, which consists of MAP kinase phosphorylation, transcriptior factor activation and the subsequent induction of various cytokines, chemokines and, importantly, smooth muscle inhibitory substances, such as nitric oxide and prostaglandins from iNOS and COX-2. This local molecular inflammatory milieu leads to leukocyte extravasation. Data suggests that the muscularis macrophage network is the conductor of the molecular and cellular inflammatory responses which causes ileus. PMID- 18402646 TI - Pathogenesis of gastro-oesophageal reflux disease and novel options for its therapy. AB - Better understanding of the mechanisms that lead to reflux disease is an important area for future research, given the very high prevalence of this problem. During the lifetime of this journal, much has been learnt about the pathophysiology of reflux disease. Abnormally, frequent acid reflux plays a key role in pathogenesis: this reflux occurs predominantly during transient lower oesophageal sphincter relaxations. Analysis of the literature suggests that the importance of transient relaxations as the major permissive event for occurrence of acid reflux is currently substantially underestimated. 'Transient relaxation' is an inexact descriptor, as this motor programme includes inhibition of the diaphragmatic hiatus and distal oesophageal body circular muscle and contraction of the oesophageal longitudinal muscle. Laxity of the diaphragmatic hiatus and hiatus hernia are probably important factors that increase the probability for acid reflux to occur during transient relaxations and in allowing strain-induced reflux episodes. The importance of straining and low basal tone of the lower oesophageal sphincter in causing abnormal reflux has probably been overestimated, but these need more investigation. High resolution manometry is the key method for acquisition of important new insights into the normal and disordered mechanics of the antireflux function of the gastro-oesophageal junction, but as yet, the potential of this technique has been tapped relatively little. In the future, improved understanding of the mechanics of the gastro-oesophageal junction should lead to improved physical antireflux procedures. Much progress has been made in defining the control of transient relaxations and this has been translated into several promising options for a new class of drug that treats reflux disease by inhibition of transient relaxations. Clinical trials on these agents appear imminent. PMID- 18402647 TI - Lower gastrointestinal functions. AB - The human colon serves to absorb water and electrolytes, store intraluminal contents until elimination is socially convenient, and salvage nutrients by bacterial metabolism of carbohydrates that have not been absorbed in the small intestine. The anorectum is responsible for fecal continence and defecation. This article is a broad perspective of the current status and a personal perspective of future challenges in understanding lower gastrointestinal functions in health and disease in humans. PMID- 18402648 TI - Psychosocial factors in functional gastrointestinal disorders: an evolving phenomenon. AB - The psychosocial aspect of functional gastrointestinal disorders have a long and complicated investigative history. Emerging from the 1930s when the observations of individual investigators and clinicians was the norm we have evolved in the last 25 years to an increasingly sophisticated era of scientific observation using standardized nosology, validated psychometric instruments and have made use of emerging technology such as brain imaging, barostat testing and other technologies. The application of the scientific method to help improve out understanding of the relationship of psychosocial factors as they relate to gastrointestinal illnesses is slowly but surely revolutionizing gastroenterology practice. It is the purpose of this paper to review the history of "Psychosomatic Gastroenterology" to review the dimensions of psychosocial factors as they relate to gastroenterology and to review the emerging technologies which are helping us to develop this knowledge. Finally we will attempt to speculate on where the field will be going in the future. PMID- 18402649 TI - Functional gastrointestinal disorders as a public health problem. AB - The functional gastrointestinal disorders (FGIDs) are a heterogeneous group of chronic conditions that are considered to be important to public health because they are remarkably common, can be disabling, and induce a major social and economic burden. Despite the Rome consensus process, the line defining true abnormality from health is as yet imprecise. Furthermore, the concept that the FGIDs have no pathological or biochemical correlates is starting to unravel, and some candidate morphometric abnormalities (e.g. duodenal eosinophilia, colonic mastocytosis) have been documented. The quality of care that patients with FGIDs receive is not well understood, despite the high volume of patients seen in primary and specialty care. There are a remarkable number of symptom-based and disease-based co-morbidities that appear to be more common in those with FGIDs that are not alone explained by coexistent somatization. Whether there is any means of preventing the development of FGIDs post-infection is untested but has vast public health implications. Defining disease severity remains key to better understanding the public health impact of FGIDs; severity is probably influenced by the intestinal and extra-intestinal symptom burden, psychological distress and the impact on quality of life, but how these factors interact remains uncertain. An important future research direction must be to quantify at what risk level it is reasonable to prescribe a medication that may be dangerous, and stratify which subsets of patients with FGIDs would qualify for such an approach using objective well validated tools (e.g. based on accurately defining gradations of severity of disease). Finally, there are some reasons to suspect that there may be an increase in mortality in FGID sufferers; for example, unnecessary surgery or invasive testing has a small but not zero associated mortality, and this should be a factor in view of the high prevalence of disease. PMID- 18402650 TI - The trials and tribulations of drug development for functional gastrointestinal disorders. AB - Functional gastrointestinal disorders (FGID) are common conditions seen in primary care and specialty practices but many affected individuals report a lack of satisfaction with available treatments. Despite the unmet need for more effective pharmacotherapy, drug development for these conditions can be challenging on many levels. This review will discuss the rationale and challenges of drug development for FGID. The reasons for engaging in drug development include that these conditions are highly prevalent, associated with a significant economic and healthcare burden, and associated with a lack of satisfaction with current therapies. The challenges include the lack of perception that FGID are legitimate disorders, the multidimensional and complex pathophysiology of FGID, the lack of a biological marker for diagnosis and treatment response, the heterogeneity of the patient population, the lack of consensus regarding the best outcome measures for clinical trials and the perceived increased risk-benefit ratio associated with drugs for FGID. Ongoing efforts are being taken to work towards a better understanding of pathophysiology, illness severity, patient reported outcome measures, and benefit : risk assessment, and towards increasing education and communication amongst patients, clinicians, investigators, industry and regulatory agencies which will hopefully help optimize drug development strategies for FGID. PMID- 18402651 TI - Abstracts of the 12th Annual Meeting of the French Society of Pharmacology and Therapeutics, the 75th Annual Meeting of the Society of Physiology, the 29th Pharmacovigilance Meeting, the 9th APNET Seminar and the 6th CHU CIC Meeting. April 9-11, 2008, Clermont-Ferrand, France. PMID- 18402652 TI - Influence of airway management strategy on "no-flow-time" during an "advanced life support course" for intensive care nurses - a single rescuer resuscitation manikin study. AB - BACKGROUND: In 1999, the laryngeal tube (VBM Medizintechnik, Sulz, Germany) was introduced as a new supraglottic airway. It was designed to allow either spontaneous breathing or controlled ventilation during anaesthesia; additionally it may serve as an alternative to endotracheal intubation, or bag-mask ventilation during resuscitation. Several variations of this supraglottic airway exist. In our study, we compared ventilation with the laryngeal tube suction for single use (LTS-D) and a bag-mask device. One of the main points of the revised ERC 2005 guidelines is a low no-flow-time (NFT). The NFT is defined as the time during which no chest compression occurs. Traditionally during the first few minutes of resuscitation NFT is very high. We evaluated the hypothesis that utilization of the LTS-D could reduce the NFT compared to bag-mask ventilation (BMV) during simulated cardiac arrest in a single rescuer manikin study. METHODS: Participants were studied during a one day advanced life support (ALS) course. Two scenarios of arrhythmias requiring defibrillation were simulated in a manikin. One scenario required subjects to establish the airway with a LTS-D; alternatively, the second scenario required them to use BMV. The scenario duration was 430 seconds for the LTS-D scenario, and 420 seconds for the BMV scenario, respectively. Experienced ICU nurses were recruited as study subjects. Participants were randomly assigned to one of the two groups first (LTS-D and BMV) to establish the airway. Endpoints were the total NFT during the scenario, the successful airway management using the respective device, and participants' preference of one of the two strategies for airway management. RESULTS: Utilization of the LTS-D reduced NFT significantly (p < 0.01). Adherence to the time frame of ERC guidelines was 96% in the LTS-D group versus 30% in the BMV group. Two participants in the LTS-D group required more than one attempt to establish the LTS-D correctly. Once established, ventilation was effective in 100%. In a subjective evaluation all participants preferred the LTS-D over BMV to provide ventilation in a cardiac arrest scenario. CONCLUSION: In our manikin study, NFT was reduced significantly when using LTS-D compared to BMV. During cardiac arrest, the LTS-D might be a good alternative to BMV for providing and maintaining a patent airway. For personnel not experienced in endotracheal intubation it seems to be a safe airway device in a manikin use. PMID- 18402653 TI - Management of SPN in France. Pathways for definitive diagnosis of solitary pulmonary nodule: a multicentre study in 18 French districts. AB - BACKGROUND: The process of diagnosis and management of solitary pulmonary nodules (SPNs) between 1 and 3 cm is not standardized. This multicentre study investigated how diagnosis of newly discovered SPNs is managed in routine practice. METHODS: We examined 11,515 radiology reports of patients undergoing chest computed tomography (CT) at all 76 radiology centres in 18 French administrative districts covering 8,220,000 people. Information on diagnostic procedures and treatment administered from discovery to definitive diagnosis of SPN was collected prospectively. RESULTS: We identified 152 cases of newly diagnosed SPNs. Follow-up was complete for 112 patients. The median number of diagnostic tests was 4 and the mean time to diagnosis was 41.4 days. Marked variability was observed in the sequence of diagnostic tests, and 8 diagnostic pathways were identified. Patients' characteristics and radiological features of SPNs influenced the number of tests performed. Referral by specialist, history of smoking and spiculated SPN predicted the performance of at least one invasive procedure (P < 0.01). Definitive diagnosis was a malignant disease in 30 patients (26%). CONCLUSION: The diagnosis of SPN is a complex process that physicians approach in markedly different ways. Implementing practice guidelines for managing the diagnosis of SPN requires clarification. PMID- 18402654 TI - A yeast three-hybrid system that reconstitutes mammalian hypoxia inducible factor regulatory machinery. AB - BACKGROUND: Several human pathologies, including neoplasia and ischemic cardiovascular diseases, course with an unbalance between oxygen supply and demand (hypoxia). Cells within hypoxic regions respond with the induction of a specific genetic program, under the control of the Hypoxia Inducible Factor (HIF), that mediates their adaptation to the lack of oxygen. The activity of HIF is mainly regulated by the EGL-nine homolog (EGLN) enzymes that hydroxylate the alpha subunit of this transcription factor in an oxygen-dependent reaction. Hydroxylated HIF is then recognized and ubiquitinilated by the product of the tumor suppressor gene, pVHL, leading to its proteosomal degradation. Under hypoxia, the hydroxylation of HIF by the EGLNs is compromised due to the lack of oxygen, which is a reaction cosubstrate. Thus, HIF escapes degradation and drives the transcription of its target genes. Since the progression of the aforementioned pathologies might be influenced by activation of HIF-target genes, development of small molecules with the ability to interfere with the HIF regulatory machinery is of great interest. RESULTS: Herein we describe a yeast three-hybrid system that reconstitutes mammalian HIF regulation by the EGLNs and VHL. In this system, yeast growth, under specific nutrient restrictions, is driven by the interaction between the beta domain of VHL and a hydroxyproline containing HIFalpha peptide. In turn, this interaction is strictly dependent on EGLN activity that hydroxylates the HIFalpha peptide. Importantly, this system accurately preserves the specificity of the hydroxylation reaction toward specific substrates. We propose that this system, in combination with a matched control, can be used as a simple and inexpensive assay to identify molecules that specifically modulate EGLN activity. As a proof of principle we show that two known EGLN inhibitors, dimethyloxaloylglycine (DMOG) and 6-chlor-3 hydroxychinolin-2-carbonic acid-N-carboxymethylamide (S956711), have a profound and specific effect on the yeast HIF/EGLN/VHL system. CONCLUSION: The system described in this work accurately reconstitutes HIF regulation while preserving EGLN substrate specificity. Thus, it is a valuable tool to study HIF regulation, and particularly EGLN biochemistry, in a cellular context. In addition, we demonstrate that this system can be used to identify specific inhibitors of the EGLN enzymes. PMID- 18402655 TI - Early acute kidney injury and sepsis: a multicentre evaluation. AB - INTRODUCTION: We conducted a study to evaluate the incidence, risk factors and outcomes associated with early acute kidney injury (AKI) in sepsis. METHODS: The study was a retrospective interrogation of prospectively collected data from the Australian New Zealand Intensive Care Society Adult Patient Database. Data were collected from 57 intensive care units (ICUs) across Australia. In total, 120,123 patients admitted to ICU for more than 24 hours from 1 January 2000 to 31 December 2005 were included in the analysis. The main outcome measures were clinical and laboratory data and outcomes. RESULTS: Of 120,123 patients admitted, 33,375 had a sepsis-related diagnosis (27.8%). Among septic patients, 14,039 (42.1%) had concomitant AKI (septic AKI). Sepsis accounted for 32.4% of all patients with AKI. For septic AKI stratified by RIFLE (risk of renal failure, injury to the kidney, failure of kidney function, loss of kidney function and end stage kidney disease) category, 38.5% of patients belonged to the risk category, 38.8% to the injury category and 22.7% to the failure category. Septic AKI patients had greater acuity of illness (P < 0.0001), lower blood pressure (P < 0.0001), higher heart rates (P < 0.0001), worse pulmonary function measures by arterial oxygen tension/fraction of inspired oxygen ratio (P < 0.0001), greater acidaemia (P < 0.0001) and higher white cell counts (P < 0.0001) compared with patients with nonseptic AKI. Septic AKI was also associated with greater severity of AKI (RIFLE category injury or failure) compared with nonseptic AKI. Septic AKI was associated with a significantly higher crude and co-variate adjusted mortality in the ICU (19.8% versus 13.4%; odds ratio 1.60, 95% confidence interval 1.5 to 1.7; P < 0.001) and in hospital (29.7% versus 21.6%; odds ratio 1.53, 95% confidence interval 1.46 to 1.60; P < 0.001) compared with nonseptic AKI. Septic AKI was associated with higher ICU and hospital mortality across all strata of RIFLE categories. Septic AKI patients had longer durations of stay in both ICU and hospital across all strata of RIFLE categories. CONCLUSION: Septic AKI is common during the first 24 hours after ICU admission. Patients with septic AKI are generally sicker, with a higher burden of illness, and have greater abnormalities in acute physiology compared with patients with nonseptic AKI. Moreover, septic AKI is independently associated with higher odds of death and longer duration of hospitalization. PMID- 18402657 TI - The effectiveness of brief personalized normative feedback in reducing alcohol related problems amongst university students: protocol for a randomized controlled trial. AB - BACKGROUND: Studies have shown that university/college students tend to have an exaggerated view of the quantities of alcohol being consumed by their peers. Making students aware of this misperception may help change behaviour and reduce problem drinking. METHODS/DESIGN: A Solomon Three Group Design will be used. There is one intervention group and two control groups, controlling separately for measurement and for intervention effects. Recruitment, consent, randomisation and data collection are all on-line. The primary outcomes are AUDIT Score, weekly consumption, perceived social norms, and alcohol related problems; secondary outcomes include alcohol expectancies and other health behaviours. DISCUSSION: This trial will provide information on the effectiveness of an on-line personalized normative feedback intervention for alcohol misuse in university students. TRIAL REGISTRATION: International Standard Randomised Controlled Trial Number: ISRCTN30784467. PMID- 18402656 TI - Hidden layers of human small RNAs. AB - BACKGROUND: Small RNA attracts increasing interest based on the discovery of RNA silencing and the rapid progress of our understanding of these phenomena. Although recent studies suggest the possible existence of yet undiscovered types of small RNAs in higher organisms, many studies to profile small RNA have focused on miRNA and/or siRNA rather than on the exploration of additional classes of RNAs. RESULTS: Here, we explored human small RNAs by unbiased sequencing of RNAs with sizes of 19-40 nt. We provide substantial evidences for the existence of independent classes of small RNAs. Our data shows that well-characterized non coding RNA, such as tRNA, snoRNA, and snRNA are cleaved at sites specific to the class of ncRNA. In particular, tRNA cleavage is regulated depending on tRNA type and tissue expression. We also found small RNAs mapped to genomic regions that are transcribed in both directions by bidirectional promoters, indicating that the small RNAs are a product of dsRNA formation and their subsequent cleavage. Their partial similarity with ribosomal RNAs (rRNAs) suggests unrevealed functions of ribosomal DNA or interstitial rRNA. Further examination revealed six novel miRNAs. CONCLUSION: Our results underscore the complexity of the small RNA world and the biogenesis of small RNAs. PMID- 18402658 TI - Substance abusers' personality disorders and staff members' emotional reactions. AB - BACKGROUND: Previous research has indicated that aggressive behaviour and DSM-IV cluster B personality disorders (PD) may be associated with professionals' emotional reactions to clients, and that cluster C PD may be associated with positive emotional reactions. METHODS: Staff members recruited from workshops completed a self-report inventory of emotional reactions to patients, the Feeling Word Checklist-58, and substance abusers completed a self-report of DSM-IV personality disorder, the DSM-IV and ICD-10 Personality Disorder Questionnaire. Correlational analysis and multiple regression analysis was used to assess the associations between personality disorders and emotional reations. RESULTS: Cluster B disorder features were associated with feeling distance to patients, and cluster C disorder features were associated with feeling helpful towards patients. Cluster A disorders had no significant impact on emotional reactions. CONCLUSION: The findings confirm clinical experiences that personality disorder features in patients with substance abuse have an impact on staff members reactions to them. These reactions should be considered in supervision of staff, and in treatment models for patients with co-morbid personality disorders and substance abuse. PMID- 18402659 TI - Perceiving molecular evolution processes in Escherichia coli by comprehensive metabolite and gene expression profiling. AB - BACKGROUND: Evolutionary changes that are due to different environmental conditions can be examined based on the various molecular aspects that constitute a cell, namely transcript, protein, or metabolite abundance. We analyzed changes in transcript and metabolite abundance in evolved and ancestor strains in three different evolutionary conditions - excess nutrient adaptation, prolonged stationary phase adaptation, and adaptation because of environmental shift - in two different strains of bacterium Escherichia coli K-12 (MG1655 and DH10B). RESULTS: Metabolite profiling of 84 identified metabolites revealed that most of the metabolites involved in the tricarboxylic acid cycle and nucleotide metabolism were altered in both of the excess nutrient evolved lines. Gene expression profiling using whole genome microarray with 4,288 open reading frames revealed over-representation of the transport functional category in all evolved lines. Excess nutrient adapted lines were found to exhibit greater degrees of positive correlation, indicating parallelism between ancestor and evolved lines, when compared with prolonged stationary phase adapted lines. Gene-metabolite correlation network analysis revealed over-representation of membrane-associated functional categories. Proteome analysis revealed the major role played by outer membrane proteins in adaptive evolution. GltB, LamB and YaeT proteins in excess nutrient lines, and FepA, CirA, OmpC and OmpA in prolonged stationary phase lines were found to be differentially over-expressed. CONCLUSION: In summary, we report the vital involvement of energy metabolism and membrane-associated functional categories in all of the evolutionary conditions examined in this study within the context of transcript, outer membrane protein, and metabolite levels. These initial data obtained may help to enhance our understanding of the evolutionary process from a systems biology perspective. PMID- 18402660 TI - Temporal trends in hepatitis B and C infection in family blood donors from interior Sindh, Pakistan. AB - BACKGROUND: Hepatitis B (HBV) and C (HCV) infections are a serious global and national public health problem. Earlier studies have reported increasing rates of hepatitis infection in Pakistan, particularly in rural areas. Pakistan has no active surveillance program to monitor trends of these infections. The objective of this study was to verify this trend in blood donors from the rural Sindh area of the country. METHODS: The study analysed the data of blood donors of interior Sindh who donated blood at JPMC blood bank from January 1, 2004 to September 15, 2007. HBsAg status was determined by using HBsAg Serodia kit and antibodies to HCV using the Detect HCV V.3 Kit. Samples repeatedly reactive for HBsAg or anti HCV were considered positive for HBV or HCV infection respectively. RESULTS: The overall seroprevalence of HBV infection among donors was 6.2 % (95% CI 5.5%-6.9%) and did not change significantly over the study period. Overall seroprevalence of HBV infection in literate blood donors was 5.7 %(95% CI 4.7%-6.8%). Prevalence decreased significantly in this group over the study period (p = 0.05). No other significant trends in seroprevalence of HBV infection were seen in the stratified analyses. The overall seroprevalence of HCV among donors was 7.5% (95% CI 6.8% 8.3%) and increased significantly over the study period from 7.2% (95% CI 5.8% 8.7%) in 2004 to 8.9% (95% CI 7.4%-10.6%) in 2007 (p = 0.02). Significant increase in seroprevalence was particularly seen in literate (p = 0.03), non first time (p = 0.01) and Sindhi speaking (p = 0.01) donors. CONCLUSION: Our study finds a steady increase in the prevalence of HCV infection in blood donors from interior Sindh between 2004 and 2007. On the contrary, decreasing prevalence of HBV was found, particularly in literate blood donors. There may be a need to have rural community-based epidemiological studies to identify the determinants of the spread of HCV infection and also those that are limiting the spread of HBV infection particularly in the literate blood donor population. PMID- 18402661 TI - Proteochemometric modeling of HIV protease susceptibility. AB - BACKGROUND: A major obstacle in treatment of HIV is the ability of the virus to mutate rapidly into drug-resistant variants. A method for predicting the susceptibility of mutated HIV strains to antiviral agents would provide substantial clinical benefit as well as facilitate the development of new candidate drugs. Therefore, we used proteochemometrics to model the susceptibility of HIV to protease inhibitors in current use, utilizing descriptions of the physico-chemical properties of mutated HIV proteases and 3D structural property descriptions for the protease inhibitors. The descriptions were correlated to the susceptibility data of 828 unique HIV protease variants for seven protease inhibitors in current use; the data set comprised 4792 protease-inhibitor combinations. RESULTS: The model provided excellent predictability (R2 = 0.92, Q2 = 0.87) and identified general and specific features of drug resistance. The model's predictive ability was verified by external prediction in which the susceptibilities to each one of the seven inhibitors were omitted from the data set, one inhibitor at a time, and the data for the six remaining compounds were used to create new models. This analysis showed that the over all predictive ability for the omitted inhibitors was Q2 inhibitors = 0.72. CONCLUSION: Our results show that a proteochemometric approach can provide generalized susceptibility predictions for new inhibitors. Our proteochemometric model can directly analyze inhibitor-protease interactions and facilitate treatment selection based on viral genotype. The model is available for public use, and is located at HIV Drug Research Centre. PMID- 18402662 TI - Prodrug converting enzyme gene delivery by L. monocytogenes. AB - BACKGROUND: Listeria monocytogenes is a highly versatile bacterial carrier system for introducing protein, DNA and RNA into mammalian cells. The delivery of tumor antigens with the help of this carrier into tumor-bearing animals has been successfully carried out previously and it was recently reported that L. monocytogenes is able to colonize and replicate within solid tumors after local or even systemic injection. METHODS: Here we report on the delivery of two prodrug converting enzymes, purine-deoxynucleoside phosphorylase (PNP) and a fusion protein consisting of yeast cytosine deaminase and uracil phosphoribosyl transferase (FCU1) into cancer cells in culture by L. monocytogenes. Transfer of the prodrug converting enzymes was achieved by bacterium mediated transfer of eukaryotic expression plasmids or by secretion of the proteins directly into the host cell cytosol by the infecting bacteria. RESULTS: The results indicate that conversion of appropriate prodrugs to toxic drugs in the cancer cells occured after both procedures although L. monocytogenes-mediated bactofection proved to be more efficient than enzyme secretion 4T1, B16 and COS-1 tumor cells. Exchanging the constitutively PCMV-promoter with the melanoma specific P4xTETP promoter resulted in melanoma cell-specific expression of the prodrug converting enzymes but reduced the efficiencies. CONCLUSION: These experiments open the way for bacterium mediated tumor specific activation of prodrugs in live animals with tumors. PMID- 18402663 TI - Rapid short-duration hypothermia with cold saline and endovascular cooling before reperfusion reduces microvascular obstruction and myocardial infarct size. AB - BACKGROUND: The aim of this study was to evaluate the combination of a rapid intravenous infusion of cold saline and endovascular hypothermia in a closed chest pig infarct model. METHODS: Pigs were randomized to pre-reperfusion hypothermia (n = 7), post-reperfusion hypothermia (n = 7) or normothermia (n = 5). A percutaneous coronary intervention balloon was inflated in the left anterior descending artery for 40 min. Hypothermia was started after 25 min of ischemia or immediately after reperfusion by infusion of 1000 ml of 4 degrees C saline and endovascular hypothermia. Area at risk was evaluated by in vivo SPECT. Infarct size was evaluated by ex vivo MRI. RESULTS: Pre-reperfusion hypothermia reduced infarct size/area at risk by 43% (46 +/- 8%) compared to post-reperfusion hypothermia (80 +/- 6%, p < 0.05) and by 39% compared to normothermia (75 +/- 5%, p < 0.05). Pre-reperfusion hypothermia infarctions were patchier in appearance with scattered islands of viable myocardium. Pre-reperfusion hypothermia abolished (0%, p < 0.001), and post-reperfusion hypothermia significantly reduced microvascular obstruction (10.3 +/- 5%; p < 0.05), compared to normothermia: (30.2 +/- 5%). CONCLUSION: Rapid hypothermia with cold saline and endovascular cooling before reperfusion reduces myocardial infarct size and microvascular obstruction. A novel finding is that hypothermia at the onset of reperfusion reduces microvascular obstruction without reducing myocardial infarct size. Intravenous administration of cold saline combined with endovascular hypothermia provides a method for a rapid induction of hypothermia suggesting a potential clinical application. PMID- 18402664 TI - Protein fingerprints of cultured CA3-CA1 hippocampal neurons: comparative analysis of the distribution of synaptosomal and cytosolic proteins. AB - BACKGROUND: All studies aimed at understanding complex molecular changes occurring at synapses face the problem of how a complete view of the synaptic proteome and of its changes can be efficiently met. This is highly desirable when synaptic plasticity processes are analyzed since the structure and the biochemistry of neurons and synapses get completely reshaped. Because most molecular studies of synapses are nowadays mainly or at least in part based on protein extracts from neuronal cultures, this is not a feasible option: these simplified versions of the brain tissue on one hand provide an homogeneous pure population of neurons but on the other yield only tiny amounts of proteins, many orders of magnitude smaller than conventional brain tissue. As a way to overcome this limitation and to find a simple way to screen for protein changes at cultured synapses, we have produced and characterized two dimensional electrophoresis (2DE) maps of the synaptic proteome of CA3-CA1 hippocampal neurons in culture. RESULTS: To obtain 2D maps, hippocampal cultures were mass produced and after synaptic maturation, proteins were extracted following subfractionation procedures and separated by 2D gel electrophoresis. Similar maps were obtained for the crude cytosol of cultured neurons and for synaptosomes purified from CA3-CA1 hippocampal tissue. To efficiently compare these different maps some clearly identifiable reference points were molecularly identified by mass spectrometry and immunolabeling methods. This information was used to run a differential analysis and establish homologies and dissimilarities in these 2D protein profiles. CONCLUSION: Because reproducible fingerprints of cultured synapses were clearly obtained, we believe that our mapping effort could represent a simple tool to screen for protein expression and/or protein localization changes in CA3-CA1 hippocampal neurons following plasticity. PMID- 18402665 TI - Minimum detectable and minimal clinically important changes for pain in patients with nonspecific neck pain. AB - BACKGROUND: The minimal detectable change (MDC) and the minimal clinically important changes (MCIC) have been explored for nonspecific low back pain patients and are similar across different cultural settings. No data on MDC and MCIC for pain severity are available for neck pain patients. The objectives of this study were to estimate MDC and MCIC for pain severity in subacute and chronic neck pain (NP) patients, to assess if MDC and MCIC values are influenced by baseline values and to explore if they are different in the subset of patients reporting referred pain, and in subacute versus chronic patients. METHODS: Subacute and chronic patients treated in routine clinical practice of the Spanish National Health Service for neck pain, with or without pain referred to the arm, and a pain severity > or = 3 points on a pain intensity number rating scale (PI NRS), were included in this study. Patients' own "global perceived effect" over a 3 month period was used as the external criterion. The minimal detectable change (MDC) was estimated by means of the standard error of measurement in patients who self-assess as unchanged. MCIC were estimated by the mean value of change score in patients who self-assess as improved (mean change score, MCS), and by the optimal cutoff point in receiver operating characteristics curves (ROC). The effect on MDC and MCIC of initial scores, duration of pain, and existence of referred pain were assessed. RESULTS: 658 patients were included, 487 of them with referred pain. MDC was 4.0 PI-NRS points for neck pain in the entire sample, 4.2 for neck pain in patients who also had referred pain, and 6.2 for referred pain. MCS was 4.1 and ROC was 1.5 for referred and for neck pain, both in the entire sample and in patients who also complained of referred pain. ROC was lower (0.5 PI-NRS points) for subacute than for chronic patients (1.5 points). MCS was higher for patients with more intense baseline pain, ranging from 2.4 to 4.9 PI NRS for neck pain and from 2.4 to 5.3 for referred pain. CONCLUSION: In general, improvements < or = 1.5 PI-NRS points could be seen as irrelevant. Above that value, the cutoff point for clinical relevance depends on the methods used to estimate MCIC and on the patient's baseline severity of pain. MDC and MCIC values in neck pain patients are similar to those for low back pain and other painful conditions. PMID- 18402666 TI - A stable isotope dual-labelling approach to detect multiple insemination in un irradiated and irradiated Anopheles arabiensis mosquitoes. AB - BACKGROUND: In the context of a Sterile Insect Technique programme, the occurrence of multiple insemination in the malaria mosquito Anopheles arabiensis Patton was studied using a novel labelling system with the stable isotopes 15N and 13C. The incidence of multiple insemination in the absence of radiation, and when males were irradiated in the pupal stage and competed against un-irradiated males were assessed. Males used in the experiments were labelled with either 15N or 13C and the label was applied to the larval rearing water. Males with either label and virgin females were caged at a 1:1:1 ratio. Males used in the radiation treatments were irradiated in the pupal stage with a partially or fully sterilizing dose of 70 or 120 Gy, respectively. After mating, females were dissected and inseminated spermathecae analysed using mass spectrometry. RESULTS: The data indicate that about 25% of inseminated females had been inseminated multiply. The presence of irradiated males in the experiments did not affect the incidence of multiple insemination. In line with previous research, irradiated males were generally less competitive than un-irradiated males. CONCLUSION: The implications of these findings for the Sterile Insect Technique are discussed, and further experiments recommended. The dual-labelling system used to determine paternity gave good results for 13C, however, for 15N it is recommended to increase the amount of label in future studies. PMID- 18402667 TI - Development, reliability and factor analysis of a self-administered questionnaire which originates from the World Health Organization's Composite International Diagnostic Interview - Short Form (CIDI-SF) for assessing mental disorders. AB - BACKGROUND: The Composite International Diagnostic Interview - Short Form consists of short form scales for evaluating psychiatric disorders. Also for this version training of the interviewer is required. Moreover, the confidentiality could be not adequately protected.This study focuses on the preliminary validation of a brief self-completed questionnaire which originates from the CIDI SF. SAMPLING AND METHODS: A preliminary version was assessed for content and face validity. An intermediate version was evaluated for test-retest reliability. The final version of the questionnaire was evaluated for factor exploratory analysis, and internal consistency. RESULTS: After the modifications by the focus groups, the questionnaire included 29 initial probe questions and 56 secondary questions. The test retest reliability weighted Kappas were acceptable to excellent for the vast majority of questions. Factor analysis revealed six factors explaining 53.6% of total variance. Cronbach's alpha was 0.89 for the questionnaire and 0.89, 0.67, 0.71, 0.71, 0.49, and 0.67, for the six factors respectively. CONCLUSION: The questionnaire has satisfactory reliability, and internal consistency, and might be efficient for using in community research and clinical practice. In the future, the questionnaire could be further validated (i.e., concurrent validity, discriminant validity). PMID- 18402668 TI - Epiphytic bacterial community composition on two common submerged macrophytes in brackish water and freshwater. AB - BACKGROUND: Plants and their heterotrophic bacterial biofilm communities possibly strongly interact, especially in aquatic systems. We aimed to ascertain whether different macrophytes or their habitats determine bacterial community composition. We compared the composition of epiphytic bacteria on two common aquatic macrophytes, the macroalga Chara aspera Willd. and the angiosperm Myriophyllum spicatum L., in two habitats, freshwater (Lake Constance) and brackish water (Schaproder Bodden), using fluorescence in situ hybridization. The bacterial community composition was analysed based on habitat, plant species, and plant part. RESULTS: The bacterial abundance was higher on plants from brackish water [5.3 x 10(7) cells (g dry mass)-1] than on plants from freshwater [1.3 x 10(7) cells (g dry mass)-1], with older shoots having a higher abundance. The organic content of freshwater plants was lower than that of brackish water plants (35 vs. 58%), and lower in C. aspera than in M. spicatum (41 vs. 52%). The content of nutrients, chlorophyll, total phenolic compounds, and anthocyanin differed in the plants and habitats. Especially the content of total phenolic compounds and anthocyanin was higher in M. spicatum, and in general higher in the freshwater than in the brackish water habitat. Members of the Cytophaga Flavobacteria-Bacteroidetes group were abundant in all samples (5-35% of the total cell counts) and were especially dominant in M. spicatum samples. Alphaproteobacteria were the second major group (3-17% of the total cell counts). Betaproteobacteria, gammaproteobacteria, and actinomycetes were present in all samples (5 or 10% of the total cell counts). Planctomycetes were almost absent on M. spicatum in freshwater, but present on C. aspera in freshwater and on both plants in brackish water. CONCLUSION: Bacterial biofilm communities on the surface of aquatic plants might be influenced by the host plant and environmental factors. Distinct plant species, plant part and habitat specific differences in total cell counts and two bacterial groups (CFB, planctomycetes) support the combined impact of substrate (plant) and habitat on epiphytic bacterial community composition. The presence of polyphenols might explain the distinct bacterial community on freshwater M. spicatum compared to that of M. spicatum in brackish water and of C. aspera in both habitats. PMID- 18402669 TI - Gene identification and protein classification in microbial metagenomic sequence data via incremental clustering. AB - BACKGROUND: The identification and study of proteins from metagenomic datasets can shed light on the roles and interactions of the source organisms in their communities. However, metagenomic datasets are characterized by the presence of organisms with varying GC composition, codon usage biases etc., and consequently gene identification is challenging. The vast amount of sequence data also requires faster protein family classification tools. RESULTS: We present a computational improvement to a sequence clustering approach that we developed previously to identify and classify protein coding genes in large microbial metagenomic datasets. The clustering approach can be used to identify protein coding genes in prokaryotes, viruses, and intron-less eukaryotes. The computational improvement is based on an incremental clustering method that does not require the expensive all-against-all compute that was required by the original approach, while still preserving the remote homology detection capabilities. We present evaluations of the clustering approach in protein-coding gene identification and classification, and also present the results of updating the protein clusters from our previous work with recent genomic and metagenomic sequences. The clustering results are available via CAMERA, (http://camera.calit2.net). CONCLUSION: The clustering paradigm is shown to be a very useful tool in the analysis of microbial metagenomic data. The incremental clustering method is shown to be much faster than the original approach in identifying genes, grouping sequences into existing protein families, and also identifying novel families that have multiple members in a metagenomic dataset. These clusters provide a basis for further studies of protein families. PMID- 18402670 TI - N-acetyltransferase 8, a positional candidate for blood pressure and renal regulation: resequencing, association and in silico study. AB - BACKGROUND: Kidneys have an important function in blood pressure (BP) regulation and elevated BP may lead to kidney failure. Chr2p12-p13 region linked to BP traits in multiple studies harbours a potential candidate for BP and renal function, N-acetyltransferase 8 (NAT8) expressed in embryonic and adult kidney and associated with nephrotoxicity response. METHODS/RESULTS: We report the first study exploring NAT8 as a potential candidate gene for blood pressure and kidney function. The resequencing (n = 42, random Estonian samples) identified 15 NAT8 polymorphisms, including 6 novel variants. The diversity of NAT8 5' upstream region (pi/bp = 0.00320) exceeded up to 10 times the variation in the NAT8 genic region (pi/bp = 0.00037) as well as the average variation (pi/bp = 0.00040) for the promoters of 29 reference genes associated with hypertension. We suggest that a potential source for such high variation could be an active gene conversion process from NAT8B duplicate gene to NAT8. Similarly to NAT8, several reference genes with the most variable upstream regions have also duplicate copies. The NAT8 promoter SNPs were targeted with pilot quantitative association studies for blood pressure (n = 137, healthy unrelated individuals) and for the index of kidney function - estimated glomerular filtration rate (eGFR; n = 157 hypertensives with and without nephropathy). Minor alleles of these polymorphisms revealed a significant protective effect against elevated systolic BP as well as kidney failure in hypertension patients (p < 0.05; linear regression model, addictive effect). CONCLUSION: The full resequencing and pilot association study of a novel positional candidate gene for blood pressure and renal function, human N-acetyltransferase 8, suggested a contribution of highly variable NAT8 promoter polymorphisms in determination of systolic blood pressure and eGFR. Based on in silico analysis, we raise the hypothesis that the alternative SNP alleles of the NAT8 upstream region may have differential effect on gene expression. PMID- 18402671 TI - Evaluating human papillomavirus vaccination programs in Canada: should provincial healthcare pay for voluntary adult vaccination? AB - BACKGROUND: Recently, provincial health programs in Canada and elsewhere have begun rolling out vaccination against human papillomavirus for girls aged 9-13. While vaccination is voluntary, the cost of vaccination is waived, to encourage parents to have their daughters vaccinated. Adult women who are eligible for the vaccine may still receive it, but at a cost of approximately CAN$400. Given the high efficacy and immunogenicity of the vaccine, the possibility of eradicating targeted types of the virus may be feasible, assuming the vaccination programs are undertaken strategically. METHODS: We develop a mathematical model to describe the epidemiology of vaccination against human papillomavirus, accounting for a widespread childhood vaccination program that may be supplemented by voluntary adult vaccination. A stability analysis is performed to determine the stability of the disease-free equilibrium. The critical vaccine efficacy and immunogenicity thresholds are derived, and the minimum level of adult vaccination required for eradication of targeted types is determined. RESULTS: We demonstrate that eradication of targeted types is indeed feasible, although the burden of coverage for a childhood-only vaccination program may be high. However, if a small, but non-negligible, proportion of eligible adults can be vaccinated, then the possibility of eradication of targeted types becomes much more favourable. We provide a threshold for eradication in general communities and illustrate the results with numerical simulations. We also investigate the effects of suboptimal efficacy and immunogenicity and show that there is a critical efficacy below which eradication of targeted types is not possible. If eradication is possible, then there is a critical immunogenicity such that even 100% childhood vaccination will not eradicate the targeted types of the virus and must be supplemented with voluntary adult vaccination. However, the level of adult vaccination coverage required is modest and may be achieved simply by removing the cost burden to vaccination. CONCLUSION: We recommend that provincial healthcare programs should pay for voluntary adult vaccination for women aged 14-26. However, it should be noted that our model results are preliminary, in that we have made a number of simplifying assumptions, including a lack of age-dependency in sexual partner rates, a lack of sexual activity outside of the vaccine age-range among females and a uniform age of sexual debut; thus, further work is desired to enhance the external generalisability of our results. PMID- 18402672 TI - Minisequencing mitochondrial DNA pathogenic mutations. AB - BACKGROUND: There are a number of well-known mutations responsible of common mitochondrial DNA (mtDNA) diseases. In order to overcome technical problems related to the analysis of complete mtDNA genomes, a variety of different techniques have been proposed that allow the screening of coding region pathogenic mutations. METHODS: We here propose a minisequencing assay for the analysis of mtDNA mutations. In a single reaction, we interrogate a total of 25 pathogenic mutations distributed all around the whole mtDNA genome in a sample of patients suspected for mtDNA disease. RESULTS: We have detected 11 causal homoplasmic mutations in patients suspected for Leber disease, which were further confirmed by standard automatic sequencing. Mutations m.11778G>A and m.14484T>C occur at higher frequency than expected by change in the Galician (northwest Spain) patients carrying haplogroup J lineages (Fisher's Exact test, P-value < 0.01). The assay performs well in mixture experiments of wild:mutant DNAs that emulate heteroplasmic conditions in mtDNA diseases. CONCLUSION: We here developed a minisequencing genotyping method for the screening of the most common pathogenic mtDNA mutations which is simple, fast, and low-cost. The technique is robust and reproducible and can easily be implemented in standard clinical laboratories. PMID- 18402673 TI - Data capture in bioinformatics: requirements and experiences with Pedro. AB - BACKGROUND: The systematic capture of appropriately annotated experimental data is a prerequisite for most bioinformatics analyses. Data capture is required not only for submission of data to public repositories, but also to underpin integrated analysis, archiving, and sharing - both within laboratories and in collaborative projects. The widespread requirement to capture data means that data capture and annotation are taking place at many sites, but the small scale of the literature on tools, techniques and experiences suggests that there is work to be done to identify good practice and reduce duplication of effort. RESULTS: This paper reports on experience gained in the deployment of the Pedro data capture tool in a range of representative bioinformatics applications. The paper makes explicit the requirements that have recurred when capturing data in different contexts, indicates how these requirements are addressed in Pedro, and describes case studies that illustrate where the requirements have arisen in practice. CONCLUSION: Data capture is a fundamental activity for bioinformatics; all biological data resources build on some form of data capture activity, and many require a blend of import, analysis and annotation. Recurring requirements in data capture suggest that model-driven architectures can be used to construct data capture infrastructures that can be rapidly configured to meet the needs of individual use cases. We have described how one such model-driven infrastructure, namely Pedro, has been deployed in representative case studies, and discussed the extent to which the model-driven approach has been effective in practice. PMID- 18402674 TI - Data-driven haemodynamic response function extraction using Fourier-wavelet regularised deconvolution. AB - BACKGROUND: We present a simple, data-driven method to extract haemodynamic response functions (HRF) from functional magnetic resonance imaging (fMRI) time series, based on the Fourier-wavelet regularised deconvolution (ForWaRD) technique. HRF data are required for many fMRI applications, such as defining region-specific HRFs, effciently representing a general HRF, or comparing subject specific HRFs. RESULTS: ForWaRD is applied to fMRI time signals, after removing low-frequency trends by a wavelet-based method, and the output of ForWaRD is a time series of volumes, containing the HRF in each voxel. Compared to more complex methods, this extraction algorithm requires few assumptions (separability of signal and noise in the frequency and wavelet domains and the general linear model) and it is fast (HRF extraction from a single fMRI data set takes about the same time as spatial resampling). The extraction method is tested on simulated event-related activation signals, contaminated with noise from a time series of real MRI images. An application for HRF data is demonstrated in a simple event related experiment: data are extracted from a region with significant effects of interest in a first time series. A continuous-time HRF is obtained by fitting a nonlinear function to the discrete HRF coeffcients, and is then used to analyse a later time series. CONCLUSION: With the parameters used in this paper, the extraction method presented here is very robust to changes in signal properties. Comparison of analyses with fitted HRFs and with a canonical HRF shows that a subject-specific, regional HRF significantly improves detection power. Sensitivity and specificity increase not only in the region from which the HRFs are extracted, but also in other regions of interest. PMID- 18402675 TI - Analyses of murine GBP homology clusters based on in silico, in vitro and in vivo studies. AB - The interactions between pathogens and hosts lead to a massive upregulation of antimicrobial host effector molecules. Among these, the 65 kDa guanylate binding proteins (GBPs) are interesting candidates as intricate components of the host effector molecule repertoire. Members of the GBP family are highly conserved in vertebrates. Previous reports indicate an antiviral activity of human GBP1 (hGBP1) and murine GBP2 (mGBP2). We recently demonstrated that distinct murine GBP (mGBP) family members are highly upregulated upon Toxoplasma gondii infection and localize around the intracellular protozoa T. gondii. Moreover, we characterised five new mGBP family members within the murine 65 kDa GBP family. Here, we identified a new mGBP locus named mGbp11. Based on bacterial artificial chromosome (BAC), expressed sequence tag (EST), and RT-PCR analyses this study provides a detailed insight into the genomic localization and organization of the mGBPs. These analyses revealed a 166-kb spanning region on chromosome 3 harboring five transcribed mGBPs (mGbp1, mGbp2, mGbp3, mGbp5, and mGbp7) and one pseudogene (pseudomGbp1), as well as a 332-kb spanning region on chromosome 5 consisting of six transcribed mGBPs (mGbp4, mGbp6, mGbp8, mGbp9, mGbp10, and mGbp11), and one pseudogene (pseudomgbp2). Besides the strikingly high homology of 65% to 98% within the coding sequences, the mGBPs on chromosome 5 cluster also exhibit a highly homologous exon-intron structure whereas the mGBP on chromosome 3 reveals a more divergent exon-intron structure. This study details the comprehensive genomic organization of mGBPs and suggests that a continuously changing microbial environment has exerted evolutionary pressure on this gene family leading to multiple gene amplifications. A list of links for this article can be found in the Availability and requirements section. PMID- 18402676 TI - Extracting expression modules from perturbational gene expression compendia. AB - BACKGROUND: Compendia of gene expression profiles under chemical and genetic perturbations constitute an invaluable resource from a systems biology perspective. However, the perturbational nature of such data imposes specific challenges on the computational methods used to analyze them. In particular, traditional clustering algorithms have difficulties in handling one of the prominent features of perturbational compendia, namely partial coexpression relationships between genes. Biclustering methods on the other hand are specifically designed to capture such partial coexpression patterns, but they show a variety of other drawbacks. For instance, some biclustering methods are less suited to identify overlapping biclusters, while others generate highly redundant biclusters. Also, none of the existing biclustering tools takes advantage of the staple of perturbational expression data analysis: the identification of differentially expressed genes. RESULTS: We introduce a novel method, called ENIGMA, that addresses some of these issues. ENIGMA leverages differential expression analysis results to extract expression modules from perturbational gene expression data. The core parameters of the ENIGMA clustering procedure are automatically optimized to reduce the redundancy between modules. In contrast to the biclusters produced by most other methods, ENIGMA modules may show internal substructure, i.e. subsets of genes with distinct but significantly related expression patterns. The grouping of these (often functionally) related patterns in one module greatly aids in the biological interpretation of the data. We show that ENIGMA outperforms other methods on artificial datasets, using a quality criterion that, unlike other criteria, can be used for algorithms that generate overlapping clusters and that can be modified to take redundancy between clusters into account. Finally, we apply ENIGMA to the Rosetta compendium of expression profiles for Saccharomyces cerevisiae and we analyze one pheromone response-related module in more detail, demonstrating the potential of ENIGMA to generate detailed predictions. CONCLUSION: It is increasingly recognized that perturbational expression compendia are essential to identify the gene networks underlying cellular function, and efforts to build these for different organisms are currently underway. We show that ENIGMA constitutes a valuable addition to the repertoire of methods to analyze such data. PMID- 18402677 TI - Risk factors for childhood obesity: shift of the entire BMI distribution vs. shift of the upper tail only in a cross sectional study. AB - BACKGROUND: Previous studies reported an increase of upper body mass index (BMI) quantiles for formula fed infants compared to breastfed infants, while corresponding mean differences were low. The aim of this study was to assess the impact of known risk factors for childhood obesity on the BMI distribution. METHODS: Data on 4,884 children were obtained at obligatory school entry health examinations in Bavaria (Germany). Exposure variables were formula feeding, maternal smoking in pregnancy, excessive TV-watching, low meal frequency, poor parental education, maternal overweight and high infant weight gain. Cumulative BMI distributions and Tukey mean-difference plots were used to assess possible shifts of BMI distributions by exposure. RESULTS: Maternal overweight and high infant weight gain shifted the entire BMI-distribution with an accentuation on upper quantiles to higher BMI values. In contrast, parental education, formula feeding, high TV consumption, low meal frequency and maternal smoking in pregnancy resulted in a shift of upper quantiles only. CONCLUSION: The single shifts among upper parts of the BMI distribution might be due to effect modification of the corresponding exposures by another environmental exposure or genetic predisposition. Affected individuals might represent a susceptible subpopulation of the exposed. PMID- 18402678 TI - MS-DOCK: accurate multiple conformation generator and rigid docking protocol for multi-step virtual ligand screening. AB - BACKGROUND: The number of protein targets with a known or predicted tri dimensional structure and of drug-like chemical compounds is growing rapidly and so is the need for new therapeutic compounds or chemical probes. Performing flexible structure-based virtual screening computations on thousands of targets with millions of molecules is intractable to most laboratories nor indeed desirable. Since shape complementarity is of primary importance for most protein ligand interactions, we have developed a tool/protocol based on rigid-body docking to select compounds that fit well into binding sites. RESULTS: Here we present an efficient multiple conformation rigid-body docking approach, MS-DOCK, which is based on the program DOCK. This approach can be used as the first step of a multi-stage docking/scoring protocol. First, we developed and validated the Multiconf-DOCK tool that generates several conformers per input ligand. Then, each generated conformer (bioactives and 37970 decoys) was docked rigidly using DOCK6 with our optimized protocol into seven different receptor-binding sites. MS DOCK was able to significantly reduce the size of the initial input library for all seven targets, thereby facilitating subsequent more CPU demanding flexible docking procedures. CONCLUSION: MS-DOCK can be easily used for the generation of multi-conformer libraries and for shape-based filtering within a multi-step structure-based screening protocol in order to shorten computation times. PMID- 18402679 TI - A high-throughput pipeline for designing microarray-based pathogen diagnostic assays. AB - BACKGROUND: We present a methodology for high-throughput design of oligonucleotide fingerprints for microarray-based pathogen diagnostic assays. The oligonucleotide fingerprints, or DNA microarray probes, are designed for identifying target organisms in environmental or clinical samples. The design process is implemented in a high-performance computing software pipeline that incorporates major algorithmic improvements over a previous version to both reduce computation time and improve specificity assessment. RESULTS: The algorithmic improvements result in significant reduction in runtimes, with the updated pipeline being nearly up to five-times faster than the previous version. The improvements in specificity assessment, based on multiple specificity criteria, result in robust and consistent evaluation of cross-hybridization with nontarget sequences. In addition, the multiple criteria provide finer control on the number of resulting fingerprints, which helps in obtaining a larger number of fingerprints with high specificity. Simulation tests for Francisella tularensis and Yersinia pestis, using a well-established hybridization model to estimate cross-hybridization with nontarget sequences, show that the improved specificity criteria yield a larger number of fingerprints as compared to using a single specificity criterion. CONCLUSION: The faster runtimes, achieved as the result of algorithmic improvements, are critical for extending the pipeline to process multiple target genomes. The larger numbers of identified fingerprints, obtained by considering broader specificity criteria, are essential for designing probes for hard-to-distinguish target sequences. PMID- 18402680 TI - Risk communication formats for low probability events: an exploratory study of patient preferences. AB - BACKGROUND: Clear communication about the possible outcomes of proposed medical interventions is an integral part of medical care. Despite its importance, there have been few studies comparing different formats for presenting probabilistic information to patients, especially when small probabilities are involved. The purpose of this study was to explore the potential usefulness of several new small-risk graphic communication formats. METHODS: Information about the likelihoods of cancer and cancer prevention associated with two hypothetical cancer screening programs were used to create an augmented bar chart, an augmented grouped icon display, a flow chart, and three paired combinations of these formats. In the study scenario, the baseline risk of cancer was 53 per 1,000 (5.3%). The risk associated with cancer screening option A was 38 per 1,000 (3.8%) and the risk associated with screening option B was 29 per 1,000 (2.9%). Both the augmented bar chart and the augmented grouped icon display contained magnified views of the differences in cancer risk and cancer prevention associated with the screening programs. A convenience sample of 29 subjects (mean age 56.4 years; 76% men) used the Analytic Hierarchy Process (AHP) to indicate their relative preferences for the six formats using 15 sequential paired comparisons. RESULTS: The most preferred format was the combined augmented bar chart + flow diagram (mean preference score 0.43) followed by the combined augmented icon + augmented bar chart format (mean preference score 0.22). The overall differences among the six formats were statistically significant: Kruskal Wallis Chi Square = 141.4, p < 0.0001. The three combined formats all had statistically significant higher preferences scores than the single format displays (p < 0.05). CONCLUSION: These findings suggest that patients may prefer combined, rather than single, graphic risk presentation formats and that augmented bar charts and icon displays may be useful for conveying comparative information about small risks to clinical decision makers. Further research to confirm and extend these findings is warranted. PMID- 18402681 TI - On the validity of area-based income measures to proxy household income. AB - BACKGROUND: This paper assesses the agreement between household-level income data and an area-based income measure, and whether or not discrepancies create meaningful differences when applied in regression equations estimating total household prescription drug expenditures. METHODS: Using administrative data files for the population of BC, Canada, we calculate income deciles from both area-based census data and Canada Revenue Agency validated household-level data. These deciles are then compared for misclassification. Spearman's correlation, kappa coefficients and weighted kappa coefficients are all calculated. We then assess the validity of using the area-based income measure as a proxy for household income in regression equations explaining socio-economic inequalities in total prescription drug expenditures. RESULTS: The variability between household-level income and area-based income is large. Only 37% of households are classified by area-based measures to be within one decile of the classification based on household-level incomes. Statistical evidence of the disagreement between income measures also indicates substantial misclassification, with Spearman's correlations, kappa coefficients and weighted kappa coefficients all indicating little agreement. The regression results show that the size of the coefficients changes considerably when area-based measures are used instead of household-level measures, and that use of area-based measures smooths out important variation across the income distribution. CONCLUSION: These results suggest that, in some contexts, the choice of area-based versus household-level income can drive conclusions in an important way. Access to reliable household level income/socio-economic data such as the tax-validated data used in this study would unambiguously improve health research and therefore the evidence on which health and social policy would ideally rest. PMID- 18402682 TI - Alternative splicing at NAGNAG acceptors in Arabidopsis thaliana SR and SR related protein-coding genes. AB - BACKGROUND: Several recent studies indicate that alternative splicing in Arabidopsis and other plants is a common mechanism for post-transcriptional modulation of gene expression. However, few analyses have been done so far to elucidate the functional relevance of alternative splicing in higher plants. Representing a frequent and universal subtle alternative splicing event among eukaryotes, alternative splicing at NAGNAG acceptors contributes to transcriptome diversity and therefore, proteome plasticity. Alternatively spliced NAGNAG acceptors are overrepresented in genes coding for proteins with RNA-recognition motifs (RRMs). As SR proteins, a family of RRM-containing important splicing factors, are known to be extensively alternatively spliced in Arabidopsis, we analyzed alternative splicing at NAGNAG acceptors in SR and SR-related genes. RESULTS: In a comprehensive analysis of the Arabidopsis thaliana genome, we identified 6,772 introns that exhibit a NAGNAG acceptor motif. Alternative splicing at these acceptors was assessed using available EST data, complemented by a sequence-based prediction method. Of the 36 identified introns within 30 SR and SR-related protein-coding genes that have a NAGNAG acceptor, we selected 15 candidates for an experimental analysis of alternative splicing under several conditions. We provide experimental evidence for 8 of these candidates being alternatively spliced. Quantifying the ratio of NAGNAG-derived splice variants under several conditions, we found organ-specific splicing ratios in adult plants and changes in seedlings of different ages. Splicing ratio changes were observed in response to heat shock and most strikingly, cold shock. Interestingly, the patterns of differential splicing ratios are similar for all analyzed genes. CONCLUSION: NAGNAG acceptors frequently occur in the Arabidopsis genome and are particularly prevalent in SR and SR-related protein-coding genes. A lack of extensive EST coverage can be compensated by using the proposed sequence-based method to predict alternative splicing at these acceptors. Our findings indicate that the differential effects on NAGNAG alternative splicing in SR and SR-related genes are organ- and condition-specific rather than gene-specific. PMID- 18402683 TI - Motives for (not) participating in a lifestyle intervention trial. AB - BACKGROUND: Non-participants can have a considerable influence on the external validity of a study. Therefore, we assessed the socio-demographic, health related, and lifestyle behavioral differences between participants and non participants in a comprehensive CVD lifestyle intervention trial, and explored the motives and barriers underlying the decision to participate or not. METHODS: We collected data on participants (n = 50) and non-participants (n = 50) who were eligible for inclusion in a comprehensive CVD lifestyle interventional trial. Questionnaires and a hospital patient records database were used to assess socio demographic, health-related and lifestyle behavioral variables. Univariate and multivariate logistic regression was used to describe the relationship between explanatory variables and study participation. Furthermore, motives and barriers that underlie study participation were investigated by means of questionnaires. RESULTS: Participants were younger, single, had a higher level of education and were employed. No statistically significant differences were found in health measures and behavioral variables. The motives for participation that were most frequently reported were: the perception of being unhealthy and willingness to change their lifestyle. The main barriers reported by non-participants were financial arguments and time investment. CONCLUSION: The differences between participants and non-participants in a lifestyle intervention trial are in mainly demographic factors. The participants consent in order to alter their lifestyle, and/or because they want to improve their health. To minimize non-participation, it is recommended that access to a lifestyle intervention program should be easy and cause no financial restraints. TRIAL REGISTRATION: ISRCTN69776211. PMID- 18402684 TI - Enhanced nigrostriatal neuron-specific, long-term expression by using neural specific promoters in combination with targeted gene transfer by modified helper virus-free HSV-1 vector particles. AB - BACKGROUND: Direct gene transfer into neurons has potential for developing gene therapy treatments for specific neurological conditions, and for elucidating neuronal physiology. Due to the complex cellular composition of specific brain areas, neuronal type-specific recombinant gene expression is required for many potential applications of neuronal gene transfer. One approach is to target gene transfer to a specific type of neuron. We developed modified Herpes Simplex Virus (HSV-1) particles that contain chimeric glycoprotein C (gC) - glial cell line derived neurotrophic factor (GDNF) or brain-derived neurotrophic factor (BDNF) proteins. HSV-1 vector particles containing either gC - GDNF or gC - BDNF target gene transfer to nigrostriatal neurons, which contain specific receptors for GDNF or BDNF. A second approach to achieve neuronal type-specific expression is to use a cell type-specific promoter, and we have used the tyrosine hydroxylase (TH) promoter to restrict expression to catecholaminergic neurons or a modified neurofilament heavy gene promoter to restrict expression to neurons, and both of these promoters support long-term expression from HSV-1 vectors. To both improve nigrostriatal-neuron specific expression, and to establish that targeted gene transfer can be followed by long-term expression, we performed targeted gene transfer with vectors that support long-term, neuronal-specific expression. RESULTS: Helper virus-free HSV-1 vector packaging was performed using either gC - GDNF or gC - BDNF and vectors that contain either the TH promoter or the modified neurofilament heavy gene promoter. Vector stocks were injected into the midbrain proximal to the substantia nigra, and the rats were sacrificed at either 4 days or 1 month after gene transfer. Immunofluorescent costaining was performed to detect both recombinant gene products and nigrostriatal neurons. The combination of targeted gene transfer with neuronal-specific promoters improved nigrostriatal neuron-specific expression (83 to 93%) compared to either approach alone, and supported long-term (1 month) expression at levels similar to those observed using untargeted gene transfer. CONCLUSION: Targeted gene transfer can be used in combination with neuronal-specific promoters to achieve a high level of nigrostriatal neuron-specific expression. Targeted gene transfer can be followed by long-term expression. Nigrostriatal neuron-specific expression may be useful for specific gene therapy approaches to Parkinson's disease or for genetic analyses of nigrostriatal neuron physiology. PMID- 18402685 TI - Immunomodulation of human monocytes following exposure to Lutzomyia intermedia saliva. AB - BACKGROUND: Sand fly saliva contains potent and complex pharmacologic molecules that are able to modulate the host's hemostatic, inflammatory, and immune systems. In this study, we evaluated the effects of salivary gland sonicate (SGS) of Lutzomyia intermedia, the natural vector of Leishmania braziliensis, on monocytes obtained from the peripheral blood mononuclear cells (PBMC) of healthy volunteers. We investigated the effects of sand fly saliva on cytokine production and surface molecule expression of LPS-stimulated human monocytes uninfected or infected with L. braziliensis. RESULTS: Pre-treatment of non-infected human monocytes with L. intermedia SGS followed by LPS-stimulation led to a significant decrease in IL-10 production accompanied by a significant increase in CD86, CD80, and HLA-DR expression. Pre-treatment with SGS followed by LPS stimulation and L. braziliensis infection led to a significant increase in TNF-alpha, IL-6, and IL-8 production without significant alterations in co-stimulatory molecule expression. However, pre-treatment with L. intermedia SGS did not result in significant changes in the infection rate of human monocytes. CONCLUSION: Our data indicate that L. intermedia saliva is able to modulate monocyte response, and, although this modulation is dissociated from enhanced infection with L. braziliensis, it may be associated with successful parasitism. PMID- 18402686 TI - Facile whole mitochondrial genome resequencing from nipple aspirate fluid using MitoChip v2.0. AB - BACKGROUND: Mutations in the mitochondrial genome (mtgenome) have been associated with many disorders, including breast cancer. Nipple aspirate fluid (NAF) from symptomatic women could potentially serve as a minimally invasive sample for breast cancer screening by detecting somatic mutations in this biofluid. This study is aimed at 1) demonstrating the feasibility of NAF recovery from symptomatic women, 2) examining the feasibility of sequencing the entire mitochondrial genome from NAF samples, 3) cross validation of the Human mitochondrial resequencing array 2.0 (MCv2), and 4) assessing the somatic mtDNA mutation rate in benign breast diseases as a potential tool for monitoring early somatic mutations associated with breast cancer. METHODS: NAF and blood were obtained from women with symptomatic benign breast conditions, and we successfully assessed the mutation load in the entire mitochondrial genome of 19 of these women. DNA extracts from NAF were sequenced using the mitochondrial resequencing array MCv2 and by capillary electrophoresis (CE) methods as a quality comparison. Sequencing was performed independently at two institutions and the results compared. The germline mtDNA sequence determined using DNA isolated from the patient's blood (control) was compared to the mutations present in cellular mtDNA recovered from patient's NAF. RESULTS: From the cohort of 28 women recruited for this study, NAF was successfully recovered from 23 participants (82%). Twenty two (96%) of the women produced fluids from both breasts. Twenty NAF samples and corresponding blood were chosen for this study. Except for one NAF sample, the whole mtgenome was successfully amplified using a single primer pair, or three pairs of overlapping primers. Comparison of MCv2 data from the two institutions demonstrates 99.200% concordance. Moreover, MCv2 data was 99.999% identical to CE sequencing, indicating that MCv2 is a reliable method to rapidly sequence the entire mtgenome. Four NAF samples contained somatic mutations. CONCLUSION: We have demonstrated that NAF is a suitable material for mtDNA sequence analysis using the rapid and reliable MCv2. Somatic mtDNA mutations present in NAF of women with benign breast diseases could potentially be used as risk factors for progression to breast cancer, but this will require a much larger study with clinical follow up. PMID- 18402687 TI - Comparison between conventional and "clinical" assessment of experimental lung fibrosis. AB - BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a treatment resistant disease with poor prognosis. Numerous compounds have been demonstrated to efficiently prevent pulmonary fibrosis (PF) in animal models but only a few were successful when given to animals with established fibrosis. Major concerns of current PF models are spontaneous resolution and high variability of fibrosis, and the lack of assessment methods that can allow to monitor the effect of drugs in individual animals over time. We used a model of experimental PF in rats and compare parameters obtained in living animals with conventional assessment tools that require removal of the lungs. METHODS: PF was induced in rats by adenoviral gene transfer of transforming growth factor-beta. Morphological and functional changes were assessed for up to 56 days by micro-CT, lung compliance (measured via a mechanical ventilator) and VO2max and compared to histomorphometry and hydroxyproline content. RESULTS: Standard histological and collagen assessment confirmed the persistent fibrotic phenotype as described before. The histomorphological scores correlated both to radiological (r2 = 0.29, p < 0.01) and functional changes (r2 = 0.51, p < 0.0001). VO2max did not correlate with fibrosis. CONCLUSION: The progression of pulmonary fibrosis can be reliably assessed and followed in living animals over time using invasive, non-terminal compliance measurements and micro-CT. This approach directly translates to the management of patients with IPF and allows to monitor therapeutic effects in drug intervention studies. PMID- 18402688 TI - Inequalities in public water supply fluoridation in Brazil: An ecological study. AB - BACKGROUND: The literature is scarce on the social and geographic inequalities in the access to and implementation of the fluoridation of public water supplies. This study adds knowledge to the Brazilian experience of the chronic privation of water and wastewater policies, access to potable water and fluoridation in the country. Thus, the aim of this study was to verify possible inequalities in the population's access to fluoridated drinking water in 246 Brazilian municipalities. METHODS: The information on the process of water fluoridation in the municipalities and in the macro region in which each municipality is located was obtained from the national epidemiological survey which was concluded in 2003. The data relating to the human development index at municipal level (HDI-M) and access to mains water came from the Brazilian Human Development Atlas, whilst the size of the population was obtained from a governmental source. The Fisher exact test (P < 0.05) was employed to identify significant associations between the explanatory variables and their ability to predict the principal outcomes of interest to this study, namely the presence or absence of the water fluoridation process in the municipalities as well as the length of time during which this measure has been implemented. Linear regression was used to observe the associations between the relevant variables in a multivariate environment. RESULTS: The results clearly showed that there is a relationship between municipalities with larger populations, located in more socio-economically advantaged regions and with better HDI-M, and where fluoridation is both present and has been implemented for a longer period of time (started before 1990). CONCLUSION: The findings suggest that the aim of treating water with fluoride may not be being adequately achieved, requiring more effective strategies so that access to this measure can be expanded equitably. PMID- 18402689 TI - Reducing the probability of false positive research findings by pre-publication validation - experience with a large multiple sclerosis database. AB - BACKGROUND: Published false positive research findings are a major problem in the process of scientific discovery. There is a high rate of lack of replication of results in clinical research in general, multiple sclerosis research being no exception. Our aim was to develop and implement a policy that reduces the probability of publishing false positive research findings. We have assessed the utility to work with a pre-publication validation policy after several years of research in the context of a large multiple sclerosis database. METHODS: The large database of the Sylvia Lawry Centre for Multiple Sclerosis Research was split in two parts: one for hypothesis generation and a validation part for confirmation of selected results. We present case studies from 5 finalized projects that have used the validation policy and results from a simulation study. RESULTS: In one project, the "relapse and disability" project as described in section II (example 3), findings could not be confirmed in the validation part of the database. The simulation study showed that the percentage of false positive findings can exceed 20% depending on variable selection. CONCLUSION: We conclude that the validation policy has prevented the publication of at least one research finding that could not be validated in an independent data set (and probably would have been a "true" false-positive finding) over the past three years, and has led to improved data analysis, statistical programming, and selection of hypotheses. The advantages outweigh the lost statistical power inherent in the process. PMID- 18402690 TI - GpnmbR150X allele must be present in bone marrow derived cells to mediate DBA/2J glaucoma. AB - BACKGROUND: The Gpnmb gene encodes a transmembrane protein whose function(s) remain largely unknown. Here, we assess if a mutant allele of Gpnmb confers susceptibility to glaucoma by altering immune functions. DBA/2J mice have a mutant Gpnmb gene and they develop a form of glaucoma preceded by a pigment dispersing iris disease and abnormalities of the immunosuppressive ocular microenvironment. RESULTS: We find that the Gpnmb genotype of bone-marrow derived cell lineages significantly influences the iris disease and the elevation of intraocular pressure. GPNMB localizes to multiple cell types, including pigment producing cells, bone marrow derived F4/80 positive antigen-presenting cells (APCs) of the iris and dendritic cells. We show that APCs of DBA/2J mice fail to induce antigen induced immune deviation (a form of tolerance) when treated with TGFbeta2. This demonstrates that some of the immune abnormalities previously identified in DBA/2J mice result from intrinsic defects in APCs. However, the tested APC defects are not dependent on a mutant Gpnmb gene. Finally, we show that the Gpnmb mediated iris disease does not require elevated IL18 or mature B or T lymphocytes. CONCLUSION: These results establish a role for Gpnmb in bone marrow derived lineages. They suggest that affects of Gpnmb on innate immunity influence susceptibility to glaucoma in DBA/2J mice. PMID- 18402691 TI - Haplotype analysis of TP53 polymorphisms, Arg72Pro and Ins16, in BRCA1 and BRCA2 mutation carriers of French Canadian descent. AB - BACKGROUND: The TP53 polymorphisms Arg72Pro (Ex4+199 G>C) and Ins16 (IVS3+24 ins16) have been proposed to modify risk of breast cancer associated with germline BRCA1 and BRCA2 mutations. Allele frequencies of these polymorphisms were investigated to determine if they modify risk in BRCA mutation carriers in breast cancer cases drawn from French Canadian cancer families, a population shown to exhibit strong founder effects. METHODS: The frequencies of the TP53 alleles, genotypes and haplotypes of 157 index breast cancer cases comprised of 42 BRCA1 mutation carriers, 57 BRCA2 mutation carriers, and 58 BRCA mutation negative cases, where each case was drawn from independently ascertained families were compared. The effect of TP53 variants on the age of diagnosis was also investigated for these groups. The TP53 polymorphisms were also investigated in 112 women of French Canadian descent with no personal history of cancer. RESULTS: The BRCA mutation-positive groups had the highest frequency of homozygous carriers of the 72Pro allele compared with mutation-negative group. The TP53 polymorphisms exhibited linkage disequilibrium (p < 0.001), where the 72Arg and Ins16minus alleles occurred in strong disequilibrium. The highest frequency of carriers of Ins16minus-72Arg haplotype occurred in the BRCA mutation-negative groups. The BRCA1 mutation carriers homozygous for the 72Pro allele had the youngest ages of diagnosis of breast cancer. However none of these observations were statistically significant. In contrast, the BRCA2 mutation carriers homozygous for the 72Pro allele had a significantly older age of diagnosis of breast cancer (p = 0.018). Moreover, in this group, the mean age of diagnosis of breast cancer in carriers of the Ins16minus-72Arg haplotype was significantly younger than that of the individuals who did not this carry this haplotype (p = 0.009). CONCLUSION: We observed no significant association of breast cancer risk with TP53 genetic variants based on BRCA1/2 mutation carrier status. Although the small sample size did not permit analysis of all possible haplotypes, we observed that BRCA2 mutation carriers harboring the Ins16minus-72Arg haplotype had a significantly younger mean age of diagnosis of breast cancer. These observations suggest that investigations in a larger French Canadian sample are warranted to further elucidate the effects of TP53 variants on age of diagnosis of breast cancer among BRCA1 and BRCA2 mutation carriers. PMID- 18402692 TI - Zebrafish brd2a and brd2b are paralogous members of the bromodomain-ET (BET) family of transcriptional coregulators that show structural and expression divergence. AB - BACKGROUND: Brd2 belongs to the bromodomain-extraterminal domain (BET) family of transcriptional co-regulators, and functions as a pivotal histone-directed recruitment scaffold in chromatin modification complexes affecting signal dependent transcription. Brd2 facilitates expression of genes promoting proliferation and is implicated in apoptosis and in egg maturation and meiotic competence in mammals; it is also a susceptibility gene for juvenile myoclonic epilepsy (JME) in humans. The brd2 ortholog in Drosophila is a maternal effect, embryonic lethal gene that regulates several homeotic loci, including Ultrabithorax. Despite its importance, there are few systematic studies of Brd2 developmental expression in any organism. To help elucidate both conserved and novel gene functions, we cloned and characterized expression of brd2 cDNAs in zebrafish, a vertebrate system useful for genetic analysis of development and disease, and for study of the evolution of gene families and functional diversity in chordates. RESULTS: We identify cDNAs representing two paralogous brd2 loci in zebrafish, brd2a on chromosome 19 and brd2b on chromosome 16. By sequence similarity, syntenic and phylogenetic analyses, we present evidence for structural divergence of brd2 after gene duplication in fishes. brd2 paralogs show potential for modular domain combinations, and exhibit distinct RNA expression patterns throughout development. RNA in situ hybridizations in oocytes and embryos implicate brd2a and brd2b as maternal effect genes involved in egg polarity and egg to embryo transition, and as zygotic genes important for development of the vertebrate nervous system and for morphogenesis and differentiation of the digestive tract. Patterns of brd2 developmental expression in zebrafish are consistent with its proposed role in Homeobox gene regulation. CONCLUSION: Expression profiles of zebrafish brd2 paralogs support a role in vertebrate developmental patterning and morphogenesis. Our study uncovers both maternal and zygotic contributions of brd2, the analysis of which may provide insight into the earliest events in vertebrate development, and the etiology of some forms of epilepsy, for which zebrafish is an important model. Knockdowns of brd2 paralogs in zebrafish may now test proposed function and interaction with homeotic loci in vertebrates, and help reveal the extent to which functional novelty or partitioning has occurred after gene duplication. PMID- 18402693 TI - A proposed approach in defining population-based rates of major injury from a trauma registry dataset: delineation of hospital catchment areas (I). AB - BACKGROUND: Determining population-based rates for major injury poses methodological challenges. We used hospital discharge data over a 10-year period (1996-2005) from a national trauma registry, the Trauma Audit and Research Network (TARN) Manchester, to construct valid numerators and denominators so that we can calculate population-based rates of major injury in the future. METHODS: We examined data from all hospitals reporting to TARN for continuity of numerator reporting; rates of completeness for patient postcodes, and clear denominator populations. We defined local market areas (>70% of patients originating from the same postcode district as the hospital). For relevant hospitals we assessed data quality: consistency of reporting, completeness of patient postcodes and for one selected hospital, North Staffordshire Royal Infirmary (NSRI), the capture rate of numerator data reporting. We used an established method based on patient flow to delineate market areas from hospitals discharges. We then assessed the potential competitors, and characterized these denominator areas. Finally we performed a denominator sensitivity analysis using a patient origin matrix based on Hospital Episodes Statistics (HES) to validate our approach. RESULTS: Sixteen hospitals met the data quality and patient flow criteria for numerator and denominator data, representing 12 hospital catchment areas across England. Data quality issues included fluctuations numbers of reported cases and poor completion of postcodes for some years. We found an overall numerator capture rate of 83.5% for the NSRI. In total we used 40,543 admissions to delineate hospital catchment areas. An average of 3.5 potential hospital competitors and 15.2 postcode districts per area were obtained. The patient origin matrix for NSRI confirmed the accuracy of the denominator/hospital catchment area from the patient flow analysis. CONCLUSION: Large national trauma registries, including TARN, hold suitable data for determining population-based injury rates. Patient postcodes from hospital discharge allow identification of denominator populations using a market area approach. PMID- 18402694 TI - Transcriptional profile of the homologous recombination machinery and characterization of the EhRAD51 recombinase in response to DNA damage in Entamoeba histolytica. AB - BACKGROUND: In eukaryotic and prokaryotic cells, homologous recombination is an accurate mechanism to generate genetic diversity, and it is also used to repair DNA double strand-breaks. RAD52 epistasis group genes involved in recombinational DNA repair, including mre11, rad50, nsb1/xrs2, rad51, rad51c/rad57, rad51b/rad55, rad51d, xrcc2, xrcc3, rad52, rad54, rad54b/rdh54 and rad59 genes, have been studied in human and yeast cells. Notably, the RAD51 recombinase catalyses strand transfer between a broken DNA and its undamaged homologous strand, to allow damaged region repair. In protozoan parasites, homologous recombination generating antigenic variation and genomic rearrangements is responsible for virulence variation and drug resistance. However, in Entamoeba histolytica the protozoan parasite responsible for human amoebiasis, DNA repair and homologous recombination mechanisms are still unknown. RESULTS: In this paper, we initiated the study of the mechanism for DNA repair by homologous recombination in the primitive eukaryote E. histolytica using UV-C (150 J/m2) irradiated trophozoites. DNA double strand-breaks were evidenced in irradiated cells by TUNEL and comet assays and evaluation of the EhH2AX histone phosphorylation status. In E. histolytica genome, we identified genes homologous to yeast and human RAD52 epistasis group genes involved in DNA double strand-breaks repair by homologous recombination. Interestingly, the E. histolytica RAD52 epistasis group related genes were differentially expressed before and after UV-C treatment. Next, we focused on the characterization of the putative recombinase EhRAD51, which conserves the typical architecture of RECA/RAD51 proteins. Specific antibodies immunodetected EhRAD51 protein in both nuclear and cytoplasmic compartments. Moreover, after DNA damage, EhRAD51 was located as typical nuclear foci-like structures in E. histolytica trophozoites. Purified recombinant EhRAD51 exhibited DNA binding and pairing activities and exchanging reactions between homologous strands in vitro. CONCLUSION: E. histolytica genome contains most of the RAD52 epistasis group related genes, which were differentially expressed when DNA double strand-breaks were induced by UV-C irradiation. In response to DNA damage, EhRAD51 protein is overexpressed and relocalized in nuclear foci-like structures. Functional assays confirmed that EhRAD51 is a bonafide recombinase. These data provided the first insights about the potential roles of the E. histolytica RAD52 epistasis group genes and EhRAD51 protein function in DNA damage response of this ancient eukaryotic parasite. PMID- 18402695 TI - Novel and nodulation-regulated microRNAs in soybean roots. AB - BACKGROUND: Small RNAs regulate a number of developmental processes in plants and animals. However, the role of small RNAs in legume-rhizobial symbiosis is largely unexplored. Symbiosis between legumes (e.g. soybean) and rhizobia bacteria (e.g. Bradyrhizobium japonicum) results in root nodules where the majority of biological nitrogen fixation occurs. We sought to identify microRNAs (miRNAs) regulated during soybean-B. japonicum symbiosis. RESULTS: We sequenced approximately 350000 small RNAs from soybean roots inoculated with B. japonicum and identified conserved miRNAs based on similarity to miRNAs known in other plant species and new miRNAs based on potential hairpin-forming precursors within soybean EST and shotgun genomic sequences. These bioinformatics analyses identified 55 families of miRNAs of which 35 were novel. A subset of these miRNAs were validated by Northern analysis and miRNAs differentially responding to B. japonicum inoculation were identified. We also identified putative target genes of the identified miRNAs and verified in vivo cleavage of a subset of these targets by 5'-RACE analysis. Using conserved miRNAs as internal control, we estimated that our analysis identified approximately 50% of miRNAs in soybean roots. CONCLUSION: Construction and analysis of a small RNA library led to the identification of 20 conserved and 35 novel miRNA families in soybean. The availability of complete and assembled genome sequence information will enable identification of many other miRNAs. The conserved miRNA loci and novel miRNAs identified in this study enable investigation of the role of miRNAs in rhizobial symbiosis. PMID- 18402696 TI - Nucleotide diversity and population differentiation of the melanocortin 1 receptor gene, MC1R. AB - BACKGROUND: The melanocortin 1 receptor gene (MC1R) is responsible for normal pigment variation in humans and is highly polymorphic with numerous population specific alleles. Some MC1R variants have been associated with skin cancer risk. RESULTS: Allele frequency data were compiled on 55 single nucleotide polymorphisms from seven geographically distinct human populations (n = 2306 individuals). MC1R nucleotide diversity, pi, was much higher (10.1 x 10-4) than in other genes for all subjects. A large degree of population differentiation, determined by FST, was also present, particularly between Asia and all other populations, due to the p.R163Q (c.488 G>A) polymorphism. The least amount of differentiation was between the United States, Northern Europe, and Southern Europe. Tajima's D statistic suggested the presence of positive selection in individuals from Europe. CONCLUSION: This study further quantifies the degree of population-specific genetic variation and suggests that positive selection may be present in European populations in MC1R. PMID- 18402698 TI - Calculating expected years of life lost for assessing local ethnic disparities in causes of premature death. AB - BACKGROUND: A core function of local health departments is to conduct health assessments. The analysis of death certificates provides information on diseases, conditions, and injuries that are likely to cause death - an important outcome indicator of population health. The expected years of life lost (YLL) measure is a valid, stand-alone measure for identifying and ranking the underlying causes of premature death. The purpose of this study was to rank the leading causes of premature death among San Francisco residents, and to share detailed methods so that these analyses can be used in other local health jurisdictions. METHODS: Using death registry data and population estimates for San Francisco deaths in 2003-2004, we calculated the number of deaths, YLL, and age-standardized YLL rates (ASYRs). The results were stratified by sex, ethnicity, and underlying cause of death. The YLL values were used to rank the leading causes of premature death for men and women, and by ethnicity. RESULTS: In the years 2003-2004, 6312 men died (73,627 years of life lost), and 5726 women died (51,194 years of life lost). The ASYR for men was 65% higher compared to the ASYR for women (8971.1 vs. 5438.6 per 100,000 persons per year). The leading causes of premature deaths are those with the largest average YLLs and are largely preventable. Among men, these were HIV/AIDS, suicide, drug overdose, homicide, and alcohol use disorder; and among women, these were lung cancer, breast cancer, hypertensive heart disease, colon cancer, and diabetes mellitus. A large health disparity exists between African Americans and other ethnic groups: African American age-adjusted overall and cause-specific YLL rates were higher, especially for homicide among men. Except for homicide among Latino men, Latinos and Asians have comparable or lower YLL rates among the leading causes of death compared to whites. CONCLUSION: Local death registry data can be used to measure, rank, and monitor the leading causes of premature death, and to measure and monitor ethnic health disparities. PMID- 18402697 TI - Reducing the false positive rate in the non-parametric analysis of molecular coevolution. AB - BACKGROUND: The strength of selective constraints operating on amino acid sites of proteins has a multifactorial nature. In fact, amino acid sites within proteins coevolve due to their functional and/or structural relationships. Different methods have been developed that attempt to account for the evolutionary dependencies between amino acid sites. Researchers have invested a significant effort to increase the sensitivity of such methods. However, the difficulty in disentangling functional co-dependencies from historical covariation has fuelled the scepticism over their power to detect biologically meaningful results. In addition, the biological parameters connecting linear sequence evolution to structure evolution remain elusive. For these reasons, most of the evolutionary studies aimed at identifying functional dependencies among protein domains have focused on the structural properties of proteins rather than on the information extracted from linear multiple sequence alignments (MSA). Non parametric methods to detect coevolution have been reported to be especially susceptible to produce false positive results based on the properties of MSAs. However, no formal statistical analysis has been performed to definitively test the differential effects of these properties on the sensitivity of such methods. RESULTS: Here we test the effect that variations on the MSA properties have over the sensitivity of non-parametric methods to detect coevolution. We test the effect that the size of the MSA (number of sequences), mean pairwise amino acid distance per site and the strength of the coevolution signal have on the ability of non-parametric methods to detect coevolution. Our results indicate that all three factors have significant effects on the accuracy of non-parametric methods. Further, introducing statistical filters improves the sensitivity and increases the statistical power of the methods to detect functional coevolution. Statistical analysis of the physico-chemical properties of amino acid sites in the context of the protein structure reveals striking dependencies among amino acid sites. Results indicate a covariation trend in the hydrophobicities and molecular weight characteristics of amino acid sites when analysing a non redundant set of 8000 protein structures. Using this biological information as filter in coevolutionary analyses minimises the false positive rate of these methods. Application of these filters to three different proteins with known functional domains supports the importance of using biological filters to detect coevolution. CONCLUSION: Coevolutionary analyses using non-parametric methods have proved difficult and highly prone to provide spurious results depending on the properties of MSAs and on the strength of coevolution between amino acid sites. The application of statistical filters to the number of pairs detected as coevolving reduces significantly the number of artifactual results. Analysis of the physico-chemical properties of amino acid sites in the protein structure context reveals their structure-dependent covariation. The application of this known biological information to the analysis of covariation greatly enhances the functional coevolutionary signal and removes historical covariation. Simultaneous use of statistical and biological data is instrumental in the detection of functional amino acid sites dependencies and compensatory changes at the protein level. PMID- 18402699 TI - Use of a Cybex NORM dynamometer to assess muscle function in patients with thoracic cancer. AB - BACKGROUND: The cachexia-anorexia syndrome impacts on patients' physical independence and quality of life. New treatments are required and need to be evaluated using acceptable and reliable outcome measures, e.g. the assessment of muscle function. The aims of this study were to: (i) examine the acceptability and reliability of the Cybex NORM dynamometer to assess muscle function in people with non-small cell lung cancer or mesothelioma; (ii) compare muscle function in this group with healthy volunteers and; (iii) explore changes in muscle function over one month. METHODS: The test consisted of 25 repetitions of isokinetic knee flexion and extension at maximal effort while seated on a Cybex NORM dynamometer. Strength and endurance for the quadriceps and hamstrings were assessed as peak torque and total work and an endurance ratio respectively. Thirteen patients and 26 volunteers completed the test on three separate visits. Acceptability was assessed by questionnaire, reliability by intraclass correlation coefficients (ICC) and tests of difference compared outcomes between and within groups. RESULTS: All subjects found the test acceptable. Peak torque and work done were reliable measures (ICC >0.80), but the endurance ratio was not. Muscle function did not differ significantly between the patient and a matched volunteer group or in either group when repeated after one month. CONCLUSION: For patients with non small cell lung cancer or mesothelioma, the Cybex NORM dynamometer provides an acceptable and reliable method of assessing muscle strength and work done. Muscle function appears to be relatively well preserved in this group and it appears feasible to explore interventions which aim to maintain or even improve this. PMID- 18402700 TI - EST Express: PHP/MySQL based automated annotation of ESTs from expression libraries. AB - BACKGROUND: Several biological techniques result in the acquisition of functional sets of cDNAs that must be sequenced and analyzed. The emergence of redundant databases such as UniGene and centralized annotation engines such as Entrez Gene has allowed the development of software that can analyze a great number of sequences in a matter of seconds. RESULTS: We have developed "EST Express", a suite of analytical tools that identify and annotate ESTs originating from specific mRNA populations. The software consists of a user-friendly GUI powered by PHP and MySQL that allows for online collaboration between researchers and continuity with UniGene, Entrez Gene and RefSeq. Two key features of the software include a novel, simplified Entrez Gene parser and tools to manage cDNA library sequencing projects. We have tested the software on a large data set (2,016 samples) produced by subtractive hybridization. CONCLUSION: EST Express is an open-source, cross-platform web server application that imports sequences from cDNA libraries, such as those generated through subtractive hybridization or yeast two-hybrid screens. It then provides several layers of annotation based on Entrez Gene and RefSeq to allow the user to highlight useful genes and manage cDNA library projects. PMID- 18402701 TI - Reliability of Ashworth and Modified Ashworth scales in children with spastic cerebral palsy. AB - BACKGROUND: Measurement of spasticity is a difficult and unresolved problem, partly due to its complexity and the fact that there are many factors involved. In the assessment of spasticity in the pediatric disabled population, methods that are easily used in practice are ordinal scales that still lack reliability. A prospective cross-sectional observational study was planned to determine the reliability of the Ashworth Scale (AS) and the Modified Ashworth Scale (MAS) in children with spastic cerebral palsy (CP). METHODS: The study included 38 children with spastic diplegic CP. The mean age for the children was 52.9 months (SD: 19.6) ranging from 18 to 108 months. The functional levels of children were classified according to the Gross Motor Function Classification System. 20 children were in Level II (52.6%), 18 were in Level III (47.4%) and 9 were in Level I (23.7%). Spasticity in hip flexors, adductors, internal rotators, hamstrings, gastrocnemius were assessed by AS and MAS. Each child was assessed by three physiotherapists in two different sessions, a week apart. The intrarater reliability was determined by paired comparison of measurements for each therapist for the two assessments. Interrater reliability was determined by paired comparisons of the three therapists' measurements on the same day. The inter and intrarater reliability of the scales were evaluated by the intraclass correlation coefficient (ICC). RESULTS: According to ICC scores, interrater reliability of AS and MAS varied from moderate to good. ICC scores of AS were between 0.54 and 0.78 and MAS were between 0.61-0.87. Test-retest results of AS and MAS varied from poor to good. ICC values were between 0.31 and 0.82 for AS and between 0.36 and 0.83 for MAS. CONCLUSION: The interrater and intrarater reliability of AS and MAS are related to muscle and joint characters. The repetition of measurements by the same physiotherapist, and experience may not affect reliability. These scales are not very reliable and assessments of spasticity using these scales should be therefore interpreted with great caution. PMID- 18402702 TI - Significance analysis of microarray for relative quantitation of LC/MS data in proteomics. AB - BACKGROUND: Although fold change is a commonly used criterion in quantitative proteomics for differentiating regulated proteins, it does not provide an estimation of false positive and false negative rates that is often desirable in a large-scale quantitative proteomic analysis. We explore the possibility of applying the Significance Analysis of Microarray (SAM) method (PNAS 98:5116-5121) to a differential proteomics problem of two samples with replicates. The quantitative proteomic analysis was carried out with nanoliquid chromatography/linear iron trap-Fourier transform mass spectrometry. The biological sample model included two Mycobacterium smegmatis unlabeled cell cultures grown at pH 5 and pH 7. The objective was to compare the protein relative abundance between the two unlabeled cell cultures, with an emphasis on significance analysis of protein differential expression using the SAM method. Results using the SAM method are compared with those obtained by fold change and the conventional t-test. RESULTS: We have applied the SAM method to solve the two sample significance analysis problem in liquid chromatography/mass spectrometry (LC/MS) based quantitative proteomics. We grew the pH5 and pH7 unlabelled cell cultures in triplicate resulting in 6 biological replicates. Each biological replicate was mixed with a common 15N-labeled reference culture cells for normalization prior to SDS/PAGE fractionation and LC/MS analysis. For each biological replicate, one center SDS/PAGE gel fraction was selected for triplicate LC/MS analysis. There were 121 proteins quantified in at least 5 of the 6 biological replicates. Of these 121 proteins, 106 were significant in differential expression by the t-test (p < 0.05) based on peptide-level replicates, 54 were significant in differential expression by SAM with Delta = 0.68 cutoff and false positive rate at 5%, and 29 were significant in differential expression by the t-test (p < 0.05) based on protein-level replicates. The results indicate that SAM appears to overcome the false positives one encounters using the peptide-based t-test while allowing for identification of a greater number of differentially expressed proteins than the protein-based t test. CONCLUSION: We demonstrate that the SAM method can be adapted for effective significance analysis of proteomic data. It provides much richer information about the protein differential expression profiles and is particularly useful in the estimation of false discovery rates and miss rates. PMID- 18402703 TI - Gene expression analyses in maize inbreds and hybrids with varying levels of heterosis. AB - BACKGROUND: Heterosis is the superior performance of F1 hybrid progeny relative to the parental phenotypes. Maize exhibits heterosis for a wide range of traits, however the magnitude of heterosis is highly variable depending on the choice of parents and the trait(s) measured. We have used expression profiling to determine whether the level, or types, of non-additive gene expression vary in maize hybrids with different levels of genetic diversity or heterosis. RESULTS: We observed that the distributions of better parent heterosis among a series of 25 maize hybrids generally do not exhibit significant correlations between different traits. Expression profiling analyses for six of these hybrids, chosen to represent diversity in genotypes and heterosis responses, revealed a correlation between genetic diversity and transcriptional variation. The majority of differentially expressed genes in each of the six different hybrids exhibited additive expression patterns, and approximately 25% exhibited statistically significant non-additive expression profiles. Among the non-additive profiles, approximately 80% exhibited hybrid expression levels between the parental levels, approximately 20% exhibited hybrid expression levels at the parental levels and ~1% exhibited hybrid levels outside the parental range. CONCLUSION: We have found that maize inbred genetic diversity is correlated with transcriptional variation. However, sampling of seedling tissues indicated that the frequencies of additive and non-additive expression patterns are very similar across a range of hybrid lines. These findings suggest that heterosis is probably not a consequence of higher levels of additive or non-additive expression, but may be related to transcriptional variation between parents. The lack of correlation between better parent heterosis levels for different traits suggests that transcriptional diversity at specific sets of genes may influence heterosis for different traits. PMID- 18402704 TI - Emergency hospital services utilization in Lleida (Spain): A cross-sectional study of immigrant and Spanish-born populations. AB - BACKGROUND: The use of emergency hospital services (EHS) has increased steadily in Spain in the last decade while the number of immigrants has increased dramatically. Studies show that immigrants use EHS differently than native-born individuals, and this work investigates demographics, diagnoses and utilization rates of EHS in Lleida (Spain). METHODS: Cross-sectional study of all the 96,916 EHS visits by patients 15 to 64 years old, attended during the years 2004 and 2005 in a public teaching hospital. Demographic data, diagnoses of the EHS visits, frequency of hospital admissions, mortality and diagnoses at hospital discharge were obtained. Utilization rates were estimated by group of origin. Poisson regression was used to estimate the rate ratios of being visited in the EHS with respect to the Spanish-born population. RESULTS: Immigrants from low income countries use EHS services more than the Spanish-born population. Differences in utilization patterns are particularly marked for Maghrebi men and women and sub-Saharan women. Immigrant males are at lower risk of being admitted to the hospital, as compared with Spanish-born males. On the other hand, immigrant women are at higher risk of being admitted. After excluding the visits with gynecologic and obstetric diagnoses, women from sub-Saharan Africa and the Maghreb are still at a higher risk of being admitted than their Spanish-born counterparts. CONCLUSION: In Lleida (Spain), immigrants use more EHS than the Spanish born population. Future research should indicate whether the same pattern is found in other areas of Spain and whether EHS use is attributable to health needs, barriers to access to the primary care services or similarities in the way immigrants access health care in their countries of origin. PMID- 18402705 TI - An intensivist-led tracheostomy review team is associated with shorter decannulation time and length of stay: a prospective cohort study. AB - INTRODUCTION: Without specific strategies to address tracheostomy care on the wards, patients discharged from the intensive care unit (ICU) with a tracheostomy may receive suboptimal care. We formed an intensivist-led multidisciplinary team to oversee ward management of such patients. To evaluate the service, we compared outcomes for the first 3 years of the service with those in the year preceding the service. METHODS: Data were prospectively collected over the course of 3 years on ICU patients not under the care of the ear, nose, and throat unit who were discharged to the ward with a tracheostomy and compared with outcomes in the year preceding the introduction of the service. Principal outcomes were decannulation time, length of stay after ICU discharge, and stay of less than 43 days (upper trim point for the disease-related group [DRG] for tracheostomy). Analysis included trend by year and multivariable analysis using a Cox proportional hazards model. P values of less than 0.05 were assumed to indicate statistical significance. As this was a quality assurance project, ethics approval was not required. RESULTS: Two hundred eighty patients were discharged with a tracheostomy over the course of a 4-year period: 41 in 2003, 60 in 2004, 95 in 2005, and 84 in 2006. Mean age was 61.8 (13.1) years, 176 (62.9%) were male, and mean APACHE (Acute Physiology and Chronic Health Evaluation) II score was 20.4 (6.4). Length of stay after ICU decreased over time (30 [13 to 52] versus 19 [10 to 34] days; P < 0.05 for trend), and a higher proportion of decannulated patients were discharged under the upper DRG trim point of 43 days (48% versus 66%; P < 0.05). Time to decannulation after ICU discharge decreased (14 [7 to 31] versus 7 [3 to 17] days; P < 0.01 for trend). Multivariate analysis showed that the hazard for decannulation increased by 24% (3% to 49%) per year. CONCLUSION: An intensivist-led tracheostomy team is associated with shorter decannulation time and length of stay which may result in financial savings for institutions. PMID- 18402706 TI - A novel representation of RNA secondary structure based on element-contact graphs. AB - BACKGROUND: Depending on their specific structures, noncoding RNAs (ncRNAs) play important roles in many biological processes. Interest in developing new topological indices based on RNA graphs has been revived in recent years, as such indices can be used to compare, identify and classify RNAs. Although the topological indices presented before characterize the main topological features of RNA secondary structures, information on RNA structural details is ignored to some degree. Therefore, it is necessity to identify topological features with low degeneracy based on complete and fine-grained RNA graphical representations. RESULTS: In this study, we present a complete and fine scheme for RNA graph representation as a new basis for constructing RNA topological indices. We propose a combination of three vertex-weighted element-contact graphs (ECGs) to describe the RNA element details and their adjacent patterns in RNA secondary structure. Both the stem and loop topologies are encoded completely in the ECGs. The relationship among the three typical topological index families defined by their ECGs and RNA secondary structures was investigated from a dataset of 6,305 ncRNAs. The applicability of topological indices is illustrated by three application case studies. Based on the applied small dataset, we find that the topological indices can distinguish true pre-miRNAs from pseudo pre-miRNAs with about 96% accuracy, and can cluster known types of ncRNAs with about 98% accuracy, respectively. CONCLUSION: The results indicate that the topological indices can characterize the details of RNA structures and may have a potential role in identifying and classifying ncRNAs. Moreover, these indices may lead to a new approach for discovering novel ncRNAs. However, further research is needed to fully resolve the challenging problem of predicting and classifying noncoding RNAs. PMID- 18402707 TI - Dental general anaesthetic receipt among Australians aged 15+ years, 1998-1999 to 2004-2005. AB - BACKGROUND: Adults receive dental general anaesthetic (DGA) care when standard dental treatment is not possible. Receipt of DGA care is resource-intensive and not without risk. This study explores DGA receipt among 15+-year-old Australians by a range of risk indicators. METHODS: DGA data were obtained from Australia's Hospital Morbidity Database from 1998-1999 to 2004-2005. Poisson regression modeling was used to examine DGA rates in relation to age, sex, Indigenous status, location and procedure. RESULTS: The overall DGA rate was 472.79 per 100,000 (95% CI 471.50-474.09). Treatment of impacted teeth (63.7%) was the most common reason for DGA receipt, followed by dental caries treatment (12.4%), although marked variations were seen by age-group. After adjusting for other covariates, DGA rates among 15-19-year-olds were 13.20 (95% CI 12.65-13.78) times higher than their 85+-year-old counterparts. Females had 1.46 (95% CI 1.45-1.47) times the rate of their male counterparts, while those living in rural/remote areas had 2.70 (95% CI 2.68-2.72) times the rate of metropolitan-dwellers. DGA rates for non-Indigenous persons were 4.88 (95% CI 4.73-5.03) times those of Indigenous persons. The DGA rate for 1+ extractions was 461.9 per 100,000 (95% CI 460.6-463.2), compared with a rate of 23.6 per 100,000 (95% CI 23.3-23.9) for 1+ restorations. CONCLUSION: Nearly two-thirds of DGAs were for treatment of impacted teeth. Persons aged 15-19 years were disproportionately represented among those receiving DGA care, along with females, rural/remote-dwellers and those identifying as non-Indigenous. More research is required to better understand the public health implications of DGA care among 15+-year-olds, and how the demand for receipt of such care might be reduced. PMID- 18402708 TI - ERCC1 and BRCA1 mRNA expression levels in metastatic malignant effusions is associated with chemosensitivity to cisplatin and/or docetaxel. AB - BACKGROUND: One of the major challenges in currently chemotherapeutic theme is lacking effective biomarkers for drug response and sensitivity. Our current study focus on two promising biomarkers, ERCC1 (excision repair cross-complementing group 1) and BRCA1 (breast cancer susceptibility gene 1). To investigate their potential role in serving as biomarkers for drug sensitivity in cancer patients with metastases, we statistically measure the mRNA expression level of ERCC1 and BRCA1 in tumor cells isolated from malignant effusions and correlate them with cisplatin and/or docetaxel chemosensitivity. METHODS: Real-time quantitative PCR is used to analysis related genes expression in forty-six malignant effusions prospectively collected from non-small cell lung cancer (NSCLC), gastric and gynecology cancer patients. Viable tumor cells obtained from malignant effusions are tested for their sensitivity to cisplatin and docetaxel using ATP-TCA assay. RESULTS: ERCC1 expression level is negatively correlated with the sensitivity to cisplatin in NSCLC patients (P = 0.001). In NSCLC and gastric group, BRCA1 expression level is negatively correlated with the sensitivity to cisplatin (NSCLC: P = 0.014; gastric: P = 0.002) while positively correlated with sensitivity to docetaxel (NSCLC: P = 0.008; gastric: P = 0.032). A significant interaction is found between ERCC1 and BRCA1 mRNA expressions on sensitivity to cisplatin (P = 0.010, n = 45). CONCLUSION: Our results demonstrate that ERCC1 and BRCA1 mRNA expression levels are correlated with in vitro chemosensitivity to cisplatin and/or docetaxel in malignant effusions of NSCLC and gastric cancer patients. And combination of ERCC1 and BRCA1 may have a better role on predicting the sensitivity to cisplatin than the single one is considered. PMID- 18402709 TI - Morphology and function of cryopreserved whole ovine ovaries after heterotopic autotransplantation. AB - BACKGROUND: The objective of this study was to perform complex characterization of cryopreserved and then autotransplanted ovaries including determination of the ability to respond to in vivo follicle stimulating hormone (FSH)-treatment, fertilizability of retrieved oocytes, and morphology, vascularization, cellular proliferation and apoptosis in sheep. METHODS: Mature crossbred ewes were divided into two groups; an intact (control) group (n = 4), and autotransplanted group (n = 4) in which oophorectomy was performed laparoscopically and ovaries with intact vascular pedicles frozen, thawed and transplanted back into the same animal at a different site. Approximately five months after autotransplantation, estrus was synchronized, ewes were treated with FSH, and ovaries were collected. For all ovaries, number of visible follicles was determined, and collected cumulus oocyte complexes (COC) were matured and fertilized in vitro. Remaining ovarian tissues were fixed for evaluation of morphology, expression of factor VIII (marker of endothelial cells), vascular endothelial growth factor (VEGF; expressed by pericytes and smooth muscle cells), and smooth muscle cell actin (SMCA; marker of pericytes and smooth muscle cells), and cellular proliferation and apoptosis. Two fully functional ovaries were collected from each control ewe (total 8 ovaries). RESULTS: Out of eight autotransplanted ovaries, a total of two ovaries with developing follicles were found. Control ewes had 10.6 +/- 2.7 follicles/ovary, oocytes were in vitro fertilized and developed to the blastocyst stage. One autotransplanted ewe had 4 visible follicles from which 3 COC were collected, but none of them was fertilized. The morphology of autotransplanted and control ovaries was similar. In control and autotransplanted ovaries, primordial, primary, secondary, antral and preovulatory follicles were found along with fully functional vascularization which was manifested by expression of factor VIII, VEGF and SMCA. Proliferating cells were detected in follicles, and the rate of apoptosis was minimal in ovaries of control and autotransplanted ovaries. CONCLUSION: These data demonstrate successful autotransplantation of a portion of frozen/thawed ovaries manifested by restoration of selected ovarian function including in vitro maturation of collected oocytes, presence of follicles from several stages of folliculogenesis and blood vessels expressing specific markers of vascularization, and proliferation and apoptosis of ovarian cells. Thus, heterotopic autotransplantation of a whole frozen/thawed ovary allows for development of preovulatory follicles, oocyte growth, and for restoration of vascularization and cellular function. However, additional improvements are required to enhance the efficiency of autotransplantation of frozen/thawed ovaries to produce more oocytes. PMID- 18402710 TI - Students' perceptions of learning environment in an Indian medical school. AB - BACKGROUND: Learning environment in any medical school is found to be important in determining students' academic success. This study was undertaken to compare the perceptions of first year and clinical phase students regarding the learning environment at Melaka Manipal Medical College (MMMC) (Manipal Campus) and also to identify the gender wise differences in their perceptions. METHODS: In the present study, the Dundee Ready Education Environment Measure (DREEM) inventory was used. DREEM was originally developed at Dundee and has been validated as a universal diagnostic inventory for assessing the quality of educational environment. In the present study, DREEM was administered to undergraduate medical students of first year (n = 118) and clinical phase (n = 108) and the scores were compared using a nonparametric test. RESULTS: Among the two batches, first year students were found to be more satisfied with the learning environment at MMMC (as indicated by their higher DREEM score) compared to the clinical batch students. Gender wise, there was not much difference in the students' perceptions. CONCLUSION: The present study revealed that both groups of students perceived the learning environment positively. Nevertheless, the study also revealed problematic areas of learning environment in our medical school which enabled us to adopt some remedial measures. PMID- 18402711 TI - A flexibly shaped space-time scan statistic for disease outbreak detection and monitoring. AB - BACKGROUND: Early detection of disease outbreaks enables public health officials to implement disease control and prevention measures at the earliest possible time. A time periodic geographical disease surveillance system based on a cylindrical space-time scan statistic has been used extensively for disease surveillance along with the SaTScan software. In the purely spatial setting, many different methods have been proposed to detect spatial disease clusters. In particular, some spatial scan statistics are aimed at detecting irregularly shaped clusters which may not be detected by the circular spatial scan statistic. RESULTS: Based on the flexible purely spatial scan statistic, we propose a flexibly shaped space-time scan statistic for early detection of disease outbreaks. The performance of the proposed space-time scan statistic is compared with that of the cylindrical scan statistic using benchmark data. In order to compare their performances, we have developed a space-time power distribution by extending the purely spatial bivariate power distribution. Daily syndromic surveillance data in Massachusetts, USA, are used to illustrate the proposed test statistic. CONCLUSION: The flexible space-time scan statistic is well suited for detecting and monitoring disease outbreaks in irregularly shaped areas. PMID- 18402713 TI - Clinically uncomplicated Plasmodium falciparum malaria with high schizontaemia: a case report. AB - BACKGROUND: The treatment options for acute Plasmodium falciparum malaria are based on the clinician classifying the patient as uncomplicated or severe according to the clinical and parasitological findings. This process is not always straightforward. CASE PRESENTATION: An adult male presented to a clinic on the western border of Thailand with a physical examination and P. falciparum trophozoite count (1.2% of infected red blood cells, IRBC) from malaria blood smear, consistent with a diagnosis of uncomplicated P. falciparum infection. However, the physician on duty treated the patient for severe malaria based on the reported P. falciparum schizont count, which was very high (0.3% IRBC), noticeably in relation to the trophozoite count and schizont:trophozoite ratio 0.25:1. On intravenous artesunate, the patient deteriorated clinically in the first 24 hours. The trophozoite count increased from 1.2% IRBC at baseline to 20.5% IRBC 18 hours following the start of treatment. By day three, the patient recovered and was discharged on day seven having completed a seven-day treatment with artesunate and mefloquine. CONCLUSION: The malaria blood smear provides only a guide to the overall parasite biomass in the body, due to the ability of P. falciparum to sequester in the microvasculature. In severe malaria, high schizont counts are associated with worse prognosis. In low transmission areas or in non immune travelers the presence of schizonts in the peripheral circulation is an indication for close patient supervision. In this case, an unusually high schizont count in a clinically uncomplicated patient was indicative of potential deterioration. Prompt treatment with intravenous artesunate is likely to have been responsible for the good clinical outcome in this case. PMID- 18402712 TI - Structure and expression pattern of Oct4 gene are conserved in vole Microtus rossiaemeridionalis. AB - BACKGROUND: Oct4 is a POU-domain transcriptional factor which is essential for maintaining pluripotency in several mammalian species. The mouse, human, and bovine Oct4 orthologs display a high conservation of nucleotide sequence and genomic organization. RESULTS: Here we report an isolation of a common vole (Microtus rossiaemeridionalis) Oct4 ortholog. Organization and exon-intron structure of vole Oct4 gene are similar to the gene organization in other mammalian species. It consists of five exons and a regulatory region including the minimal promoter, proximal and distal enhancers. Promoter and regulatory regions of the vole Oct4 gene also display a high similarity to the corresponding regions of Oct4 in other mammalian species, and are active during the transient transfection within luciferase reporter constructs into mouse P19 embryonic carcinoma cells and TG-2a embryonic stem cells. The vole Oct4 gene expression is detectable starting from the morula stage and until day 17 of embryonic development. CONCLUSION: Genomic organization of this gene and its intron-exon structure in vole are identical to those in all previously studied species: it comprises five exons and the regulatory region containing several conserved elements. The activity of the Oct4 gene in vole, as well as in mouse, is confined only to pluripotent cells. PMID- 18402714 TI - The multi-faceted assessment of independence in patients with rheumatoid arthritis: preliminary validation from the ATTAIN study. AB - OBJECTIVE: To consider the feasibility of assessing multiple facets of independence in rheumatoid arthritis (RA) using a measure developed from existing items and examining its face validity, construct validity and responsiveness to change. METHODS: The ATTAIN (Abatacept Trial in Treatment of Anti-tumor necrosis factor [TNF] Inadequate responders) database was used. Patients with RA were randomized 2:1, abatacept (n = 258) and placebo (n = 133). A multi-faceted scale to measure physical and psychosocial independence was constructed using items from the Health Assessment Questionnaire (HAQ) and Short Form 36 Health Survey (SF-36). Questions assessing activity limitations and need for outside caregiver help were also examined. Interviews with 20 RA patients assessed face validity. RESULTS: Item Response Theory analysis yielded two traits - 'Psychosocial Independence', derived from the number of days with activity limitations plus the Role Emotional, Social Functioning and Role Physical subscale items from the SF 36; and 'Physical Independence', derived from 15 HAQ items assessing need for help from another. The two traits showed no significant differential item functioning for age or gender and demonstrated good face validity. Changes over 169 days on Psychosocial Independence were greater (mean 0.46 units, 95% confidence interval [CI]: 0.17-0.75) for the abatacept group than for placebo (p = 0.002). Changes in Physical Independence were greater (mean 0.59 units, 95% CI: 0.35-0.82) for the abatacept group than for placebo (p < 0.001). CONCLUSIONS: The multi-faceted assessment of independence in RA based on items from commonly used instruments is feasible suggesting promise for evaluating independence in future clinical trials. This approach demonstrated good face and construct validity and responsiveness in RA patients who had previously failed anti-TNF therapy. However, we caution against an interpretation that these data suggest that abatacept improves independence because the component parts of this assessment came from instruments used in the ATTAIN trial where data had been previously analyzed. PMID- 18402716 TI - Anticoagulation in atrial fibrillation: the trials and tribulations of keeping within therapeutic range. AB - Atrial fibrillation (AF) is a growing epidemic in the developed world that has attracted much attention due to the associated risk of stroke and thromboembolism. This article discusses the application of oral anticoagulation (OAC) therapy with vitamin K antagonists (VKA) in order to reduce these risks. The article provides an overview of OAC management and highlights the need for greater efforts to increase time within therapeutic range. PMID- 18402715 TI - Effect of setting, monitoring intensity and patient experience on anticoagulation control: a systematic review and meta-analysis of the literature. AB - OBJECTIVES: To investigate the relationship between time spent in the recommended target International Normalised Ratio (INR) range and the setting and intensity of anti coagulant monitoring, in both treatment-experienced and treatment-naive atrial fibrillation (AF) patients receiving oral anticoagulation (OAC) therapy for the prevention of ischaemic stroke. RESEARCH DESIGN AND METHODS: Systematic review of published studies on participants with atrial fibrillation on anticoagulation therapy. We compared frequent monitoring (well-controlled, according to a strict protocol) versus infrequent monitoring (frequency representative of routine clinical practice), specialised care versus usual care, and naive versus prior anticoagulant use. Meta-analysis was performed using a random effects model. RESULTS: 36 studies were included, 22 (primary data) of AF patients managed in line with the consensus guidelines target INR range of 2.0 3.0, and 14 studies (secondary data) of mixed patient groups, including AF, with an INR target of 2.0-3.5. Both data sets were combined for sensitivity analysis. Pooled mean time in INR range was 59.1% (95% CI: 55.5, 62.8%) and 64.3% (95% CI: 60.5, 68.0%) for infrequent monitoring and frequent monitoring, respectively. Significantly more time was spent in range in specialist care settings compared to usual care: +11.3% (95% CI: 0.1-21.7%). Naive OAC users spent less time in range 56.5% (95% CI: 45.5-67.5%) than existing users 61.2% (95% CI: 57.2-65.2%). All of these differences were found to be significant in the sensitivity analyses. CONCLUSIONS: INR control is variable and dependent on monitoring intensity and duration of anticoagulant therapy. PMID- 18402717 TI - Brimonidine purite 0.1% versus brinzolamide 1% as adjunctive therapy to latanoprost in patients with glaucoma or ocular hypertension. AB - OBJECTIVE: To evaluate the efficacy and tolerability of brimonidine purite 0.1% in comparison to brinzolamide 1% when used as adjunctive therapy to latanoprost 0.005% in patients with glaucoma or ocular hypertension. METHODS: Randomized, single-center, investigator-masked, parallel-group clinical study. Patients with IOP >or= 18 mmHg while on once-daily latanoprost were randomized to adjunctive treatment with brimonidine purite TID (n = 20) or brinzolamide TID (n = 20) for 3 months. Intraocular pressure (IOP) was measured at 8 a.m., 10 a.m., and 4 p.m. at latanoprost-treated baseline and after 1 and 3 months of latanoprost and adjunctive therapy. A patient questionnaire was administered to evaluate the tolerability of eye drop instillation. RESULTS: Baseline mean diurnal IOP (+/- standard deviation, mmHg) on latanoprost was comparable between groups (brimonidine purite: 19.6 +/- 2.94; brinzolamide: 19.8 +/- 3.25; p = 0.846). Mean diurnal IOP at Month 3 was 16.3 +/- 2.63 mmHg with brimonidine purite and 17.8 +/ 2.19 mmHg with brinzolamide (p = 0.028). Adjunctive use of brimonidine purite provided greater IOP lowering than brinzolamide at 10 a.m. (p < 0.001) and 4 p.m. (p = 0.050) and equivalent IOP lowering to brinzolamide at 8 a.m. (p = 0.716). Blurred vision at Month 1 and bitter taste at Months 1 and 3 were more common upon instillation of brinzolamide eye drops. CONCLUSION: Brimonidine purite 0.1% provided significantly lower IOP compared with brinzolamide 1% when used as adjunctive therapy to latanoprost. Both adjunctive therapies were well tolerated. Limitations of this study include the use of a single site and the sample size. Additional studies are needed to further evaluate these drugs as adjunctive therapy to prostaglandin analogs. PMID- 18402718 TI - TLC of alkaloids on cyanopropyl bonded stationary phases. Part II. Connection with RP18 and silica plates. AB - Some standards of the alkaloids and synthetic or natural mixtures are separated by two-dimensional thin-layer chromatography (TLC) on different adsorbent layers. Normal- and reversed-phase systems are used to obtain significant differences in the separation selectivity. Optimization of the one-dimensional TLC separation of the alkaloids' standards is performed on cyanopropyl-silica, RP18W, and silica layers in various eluents containing (besides diluent and modifier) silanol blockers, such as diethyl amine or ammonia. The most selective systems are used for the separation of the alkaloids' mixtures by two-dimensional TLC with an adsorbent gradient method. The mixtures of alkaloids or plant extracts (Chelidonium majus, Fumaria officinalis, or Glaucium flavum) are chromatographed in system I; the plates are connected with the plate pre-coated with various adsorbent, and partly separated fractions are transferred to the second layer and developed in system II. CN-silica-RP18W and CN-silica-silica are used as the connected layers. The alkaloids are identified by R(F) values of standards, and the components of plant extracts are identified in both systems, and by the comparison of UV spectra obtained in diode array detector densitometry. PMID- 18402719 TI - Silica thin-layer chromatographic studies of surfactants with mixed aqueous organic eluents containing thiourea: simultaneous separation of co-existing cetyltrimethylammonium bromide, dodecyltrimethylammonium bromide, and polyoxyethylene (20) sorbitan monolaurate. AB - Silica thin-layer chromatography of three surfactants using various solvent systems is described. The mutual separation of coexisting cetyltrimethylammonium bromide (CTAB), dodecyltrimethyl-ammonium bromide (DTAB), and polyoxyethylene (20) sorbitan monolaurate (Tween 20) is achieved on silica layer using 5% aqueous thiourea-acetone-methanol (60:20:20,v/v/v) as the mobile phase. The effect of the carbon chain length of alcohols (methanol, ethanol, n-propanol, and n-butanol) on the mobility of these surfactants is examined on silica layers. The comparative study is performed with sulfur- (thiourea) and oxygen- (urea) containing compounds in the eluent on the mobility as well as on the separation of co existing CTAB, DTAB, and Tween 20. The interference on the resolution of the mixture of CTAB, DTAB, and Tween 20, due to presence of metal cations as impurities, is also examined. The limits of detection of CTAB, DTAB, and Tween 20 are estimated. PMID- 18402720 TI - HPTLC determination of rabeprazole and domperidone in capsules and its validation. AB - This paper describes a validated high-performance thin-layer chromatography (HPTLC) method for simultaneous estimation of rabeprazole (RA) and domperidone (DO) in pure powder and in capsule formulations. An HPTLC method separation is achieved on an aluminum sheet of silica gel 60F(254) using ethyl acetate-methanol benzene-acetonitrile (30:20:30:20 v/v) as mobile phase. Quantitation is achieved with UV detection at 287 nm over a concentration range of 400-1200 ng/spot and 600-1800 ng/spot with mean recovery of 99.82 +/- 0.74 and 99.43 +/- 0.68 for RA and DO, respectively, in the HPTLC method. This method is simple, precise, and sensitive, and it is applicable for the simultaneous determination of RA and DO in pure powder and in capsule formulation. PMID- 18402721 TI - Acid modified diatomaceous earth--a sorbent material for thin layer chromatography. AB - Natural diatomaceous earth (DE) is modified by flux calcination and refluxing with acid. To characterize natural DE, modified DE's [flux calcinated (FC)DE and FCDE-I] and silica gel 60GF(254) (Si-60GF(254)) are analyzed microscopically, physically, and chemically by various techniques. FCDE-I and Si-60GF(254) are investigated for their usefulness in the stationary phase of thin layer chromatography (TLC) both individually and in composition. Sodium diethyldithiocarbamate (DEDTC) and ammonium pyrrolidinedithiocarbamate (PyDTC) are prepared as Co or Cu (M) complexes [M(DEDTC)(2) and M(PyDTC)(2), respectively]. These complexes and their mixtures are run on thin layers of Si 60GF(254) and FCDE-I individually, and on various FCDE-I and Si-60GF(254) mixtures. Pure toluene and various toluene-cyclohexane mixtures (3:1, 1:1, 1:2, 1:3, v/v) are used as mobile phases for the running the complexes. The best analytical separations of both M(DEDTC)(2) and M(PyDTC)(2) complexes are obtained when using pure toluene and toluene-cyclohexane (3:1, 1:1, v/v) as mobile phases on FCDE-I-Si-60GF(254) (1:3, 1:1, w/w) layers as stationary phases. This study shows that it is possible to qualitatively analyze and to satisfactorily separate a mixture Cu(2+) and Co(2+) cations on cited chromatographic systems. PMID- 18402722 TI - TLC analysis of intermediates arising during the preparation of oxime HI-6 dimethanesulfonate. AB - In this study, several thin-layer chromatography (TLC) methods are described for the identification of quaternary and non-quaternary compounds (parent compounds, intermediates, by-products, and products) arisen within the synthesis of the acetylcholinesterase reactivator HI-6, at present the most promising antidote in the case of nerve agent poisonings. Using the TLC technique, particular E and Z isomers of this compound on the oxime group are separated. These TLC methods could be of high interest as quick purity control for those who are interested in development of new acetylcholinesterase reactivators and the synthesis of HI-6 in laboratories or in large-scale production. PMID- 18402723 TI - Separation and quantitation of jasmonic acid using HPTLC. AB - A rapid, simple, and stringent protocol for the detection and quantitation of jasmonic acid (JA) is designed using high-performance thin-layer chromatography. Acidified culture filtrate of Lasiodiplodia theobromae is extracted with an equal volume of ethyl acetate and spotted on silica gel 60 F(254) foil using Linomat-5 spray-on applicator. Standard JA is also spotted either internally or adjacent to the sample, and the foils are developed with isopropanol-ammonia-water [10:1:1 (v/v)] as the mobile phase. A quantitative estimation of the separated JA is performed by measuring the absorbance at 295 nm in the reflective mode. The sensitivity of the method is improved by adding internal standard to obtain a detection limit of 1 microg. The limit of quantitation is found to be 80 microg with this method. The method is shown to have selectivity, accuracy, precision, and high sample throughput, making it useful for the routine analysis of JA in basic science and perfumery industries. PMID- 18402724 TI - Implementation of solid-phase microextraction with micellar desorption method for priority phenolic compound determination in natural waters. AB - Eleven phenolic compounds considered by the Environmental Protection Agency to be priority pollutants are extracted and determined in different water samples. The method involves the extraction and clean-up step of target compounds by solid phase microextraction and micellar desorption (SPME-MD) and a second step of determination by liquid chromatography with diode array detection. Different fibers and surfactants are evaluated for the analysis of these target analytes in water samples. In the optimum conditions for the SPME process, recoveries for the target compounds are between 80% and 109%; relative standard deviations are lower than 10%, and detection limits are in the range 0.3-3.5 ng/mL. The main advantages of this method are the combination of time and efficiency, safety, and an environmentally friendly process for sample extraction prior to instrumental determination. This demonstrates that SPME-MD can be used as an alternative to traditional methods for the extraction and determination of priority phenolic compounds in natural waters from different origins. PMID- 18402725 TI - A comparison of results obtained using liquid injection and headspace solid-phase microextraction for crude oil analysis by GC with mass spectrometer detection. AB - Gas chromatographic (GC) analysis in solution and head space solid-phase microextraction (SPME)-GC analysis of a sample of crude oil gave different results. The SPME technique allowed the identification of a larger number of components than by using usual GC-mass spectrometry (MS). The method failed within the range of C14-C25 where GC-MS in solution allowed to obtain more representative results; on the contrary, SPME allowed to obtain data on the presence of volatile compounds that can not be identified in GC-MS analysis in solution. Furthermore, in the range C8-C12, SPME allowed to identify approximately 30 compounds not shown in the GC-MS analysis in solution. SPME analysis showed the presence of some alkenes not identified in GC-MS analysis in solution. SPME-GC-MS can be used in the analysis of crude oil in contaminated soil. PMID- 18402726 TI - Photodegradation of crude oil: liquid injection and headspace solid-phase microextraction for crude oil analysis by gas chromatography with mass spectrometer detector. AB - The fate of crude oil under irradiation is studied. After UV irradiation, the fraction present in the highest percentage shifts from the C8-C9 fractions to C13, using gas chromatography-mass spectrometry (GC-MS) analysis in solution. An increase of the relative amount of the C13-C25 fraction is observed, while a decrease in the relative amount of the C7-C12 fractions is present. In headspace solid-phase microextraction (HS-SPME) analysis, the C8-C10 fractions represent 53% of all the compounds detected. A decrease in the relative amount of the C8 C10 fractions is observed, while C11-C15 fractions increase. The irradiation of crude oil with a solar simulator gives a mixture the analysis of which using GC MS in solution furnishes the same type of results: the relative amounts of linear alkanes and aromatic compounds increase, while a sharp decrease in the relative amounts of branched and cyclic alkanes is observed. In the SPME analysis, a decreased relative amount of branched alkanes and alkenes, and an increase in the relative amounts of cyclic alkanes and aromatic compounds are observed. Analysis of the distribution of the compounds in all the types of compound shows that a dynamic equilibrium between different compounds and different types of compounds is present. To confirm the presence of a dynamic equilibrium, the irradiation of methylcyclohexane in the presence of 2-methylnaphthalene shows the presence in the reaction mixture of a small amount of tetradecane. PMID- 18402727 TI - Simultaneous determination of five fat-soluble vitamins in feed by high performance liquid chromatography following solid-phase extraction. AB - A high-performance liquid chromatography method is developed for the simultaneous determination of menadione, retinyl acetate, cholecalciferol, alpha-tocopherol, and alpha-tocopherol acetate in feed. The present study uses an enzyme to destroy the coating film, ethanol to extract free vitamins, and Oasis HLB cartridges to purify. Vitamins are separated using an Atlantis dC18 column. The mobile phase is methanol-water (98:2 v/v). Detection is performed with a UV-vis detector at 230 and 265 nm. The linearity, accuracy, and repeatability of this method are all satisfactory. Application of the method is suitable for the determination of the fat-soluble vitamins in general feed. PMID- 18402728 TI - Determination of closantel residues in milk and animal tissues by HPLC with fluorescence detection and SPE with oasis MAX cartridges. AB - A liquid chromatographic method for the determination of closantel residues in milk and tissues is developed and validated. An acetonitrile-acetone solution (80:20, v/v) is used for the extraction of closantel residues from milk and animal tissues, and the extract is purified by solid-phase extraction with Oasis MAX cartridges and a mixture of formic acid-acetonitrile (5:95, v/v) as the elution solution. A C(18) bonded silica column is used for chromatographic separation. The mobile phase consists of acetonitrile-water (85:15, v/v) containing 0.05% triethylamine at pH 2.5, adjusted with phosphoric acid with the flow-rate set at 1.0 mL/min. Using the fluorescence emission of closantel at lambda(ex) = 335 nm and lambda(ex) = 510 nm, the calibration curve is linear, with a correlation coefficient of 0.9999 over the concentration range of 10-5000 microg/kg for the tissue sample and 10-5000 microg/L for the milk sample. The detection limit (s/n = 3) is 3 microg/kg for tissue sample and 3 microg/L for milk sample. The intra- and inter-day repeatabilities are between 3.35-7.66% and 4.04-8.67%, respectively. The proposed method enables the quantitative determination of closantel residues at levels as low as 10 microg/kg in animal tissue samples and 10 microg/L in milk samples. PMID- 18402729 TI - Development and validation of an LC-MS assay for finasteride and its application to prostate cancer prevention trial sample analysis. AB - An analytical method is developed and validated for the quantitative determination of finasteride, a potent 5 alpha-reductase inhibitor, in human plasma. Calibration curves are linear in the concentration range of 1 to 100 ng/mL. Sample pretreatment involves a liquid-liquid extraction with ethyl acetate using 0.2 mL aliquots of plasma. Finasteride and the internal standard (beclomethasone) are separated on a Waters Symmetry Shield RP18 column (50 x 2.1 mm, 3.5 microm) and eluted using a gradient mobile phase composed of acetonitrile and 10mM ammonium acetate with 0.1% formic acid. The column eluant is monitored by mass spectrometry with electrospray ionization. A complete validation of the method is performed. For quality control samples at three different concentrations that were analyzed in quintuplicate, on six separate occasions, the accuracy and precision range from 95.2% to 101% and 3.4% to 7.3%, respectively. The developed method is subsequently applied to measure the steady state finasteride concentration of patients who participated in the Prostate Cancer Prevention Trial. PMID- 18402730 TI - Prospecting for Camptothecines from Nothapodytes nimmoniana in the Western Ghats, South India: identification of high-yielding sources of camptothecin and new families of camptothecines. AB - Camptothecin (CPT), a monoterpene alkaloid, is an important anti-cancer compound obtained from several plant sources including Camptotheca acuminta (from China) and Nothapodytes nimmoniana (from India). Currently, by far the highest levels of CPT (approximately 0.3% w/w) are reported from Nothapodytes nimmoniana, a small tree distributed in the Western Ghats, India. In recent years because of the heavy demand, there has been a serious threat of extinction of the populations of the tree in the Western Ghats forest of south India. Several studies have chemically profiled populations of the species in the Western Ghats to identify sources of high yield and therefore to enable the sustainable production and harvesting of CPT. In this study, using both high-performance liquid chromatography and liquid chromatography-mass spectrometry, we report for the first time the identification of trees that produce at least 5- to 8-fold more CPT than hitherto reported. Furthermore, we show for the first time the production of a few minor camptothecines, including 10-hydroxy camptothecin, in the stem and root bark extracts of the tree. These results have important implications for not only harnessing the high-yielding individuals for clonal multiplication but also for exploiting some of the minor camptothecines, which also have been shown to have important anti-cancer and anti-viral activity. PMID- 18402731 TI - Immediate impact of an intensive one-week laparoscopy training program on laparoscopic skills among postgraduate urologists. AB - INTRODUCTION: Laparoscopic techniques are difficult to master, especially for surgeons who did not receive such training during residency. To help urologists master challenging laparoscopic skills, a unique 5-day mini-residency (M-R) program was established at the University of California, Irvine. The first 101 participants in this program were evaluated on their laparoscopic skills acquisition at the end of the 5-day experience. METHODS: Two urologists are accepted per week into 1 of 4 training modules: (1) ureteroscopy/percutaneous renal access; (2) laparoscopic ablative renal surgery; (3) laparoscopic reconstructive renal surgery; and (4) robot-assisted prostatectomy. The program consists of didactic lectures, pelvic trainer and virtual reality simulator practice, animal and cadaver laboratory sessions, and observation or participation in human surgeries. Skills testing (ST) simulating open, laparoscopic, and robotic surgery is assessed in all of the M-R participants on training days 1 and 5. Tests include ring transfer, suture threading, cutting, and suturing. Performance is evaluated by an experienced observer using the Objective Structured Assessment of Technical Skill (OSATS) scoring system. Statistical methods used include the paired sample t test and analysis of variance at a confidence level of P65 years old. The patient demographics, comorbidities, hernia characteristics, and operative and postoperative data were compared. RESULTS: Younger patients were found to have a significantly increased BMI, while the older group had an increased number of comorbidities. No difference was found in the complication or recurrence rates between the 2 groups. CONCLUSION: Elective laparoscopic ventral hernia repair in senior citizens is safe and feasible in our experience. We believe that the decision to perform an elective hernia repair in this patient population should be based on the general condition of the patient rather than the patient's chronological age. PMID- 18402739 TI - Seroma and recurrence in laparoscopic ventral hernioplasty. AB - BACKGROUND: Recurrence after laparoscopic ventral hernioplasty is a severe problem despite surgeons' increased experience in recent years. It is well known that the main reasons for recurrences are lack of experience, bad technique, infection, and seroma. The aim of this study was to investigate the events, what caused recurrences, and the technique to prevent recurrence in laparoscopic ventral hernioplasty. METHODS: From May 1996 through December 2005, 78 patients who underwent 80 laparoscopic ventral hernioplasties (67 incisional hernias, 8 large epigastric, 5 large umbilical) were separated into 2 groups. Group A (n=28): ePTFE dual mesh patch secured intraperitoneally by full-thickness stitches and endoscopic tacks to cover the hernia defect and to overlap healthy margins by at least 2.5 cm (n=17, subgroup A1) or 4 cm (n=11, subgroup A2). In subgroup A2, a full-thickness suture was placed in the center of the hernia defect to reduce the "dead space." Group B (n=52): The same technique as in group A, but the hernia sac was cauterized by monopolar cautery (n=5) or Harmonic scalpel (n=47). The overlapping healthy margins were at least 2.5 cm (n=16, subgroup B1) or 4 cm (n=36, subgroup B2). In subgroup B2, a full-thickness suture was placed in the center of the hernia defect to reduce the dead space. Postoperatively, CT-scans were used to confirm complications or recurrences. RESULTS: In group A, 7 seromas [4 clinical (A1) and 3 subclinical (A1=1, A2=2)], 3 hematomas (A1=2, A2=1), 2 infections (A1), and 3 recurrences (10.7%) were observed (A1=2 or 11.8%, A2=1 or 9%). Two recurrences were observed in symptomatic seromas (subgroup A1) and 1 in a patient without seroma (subgroup A2). In group B, 1 subclinical seroma, 1 hematoma, and 1 recurrence (6.2%) were noted in subgroup B1. In subgroup B2, no recurrence was observed. Significantly fewer total seromas occurred in group B compared with group A (P=0.004). The total recurrence rate in group B was 1.95% (NS vs group A), but a significant difference was observed between subgroups A1 and B2 (P=0.036). CONCLUSION: Cauterization of the hernia sac and a central full-thickness suture to reduce dead space seems to prevent seroma. This technique combined with a large patch to cover at least 4 cm of healthy margins and the surgeon's experience may be sufficient to prevent recurrences in laparoscopic ventral hernioplasty. PMID- 18402740 TI - Laparoscopic evaluation of abdominal adhesions with different prosthetic meshes in rabbits. AB - BACKGROUND: The use of prosthetic materials to reinforce the abdominal wall is associated with a low index of recurrence; however, intraperitoneal placement of a foreign body may lead to adhesions. The present investigation was designed to determine adhesion formation with commercially available meshes implanted laparoscopically in rabbits. METHODS: Three different meshes were implanted laparoscopically in 24 rabbits: polypropylene (mesh A), polypropylene and sodium hyaluronate-carboxymethylcellulose (mesh B), and polypropylene and expanded polytetrafluoroethylene (mesh C). Sites of implantation for each mesh (the left lower quadrant, right lower quadrant, and lower midline) were randomly determined so that every rabbit had all 3 meshes implanted. All animals underwent diagnostic laparoscopy after 28 days to grade adhesions and histological analysis of inflammation. RESULTS: Adhesions were noticed in 46 of the 72 meshes implanted (64%). The number of adhesions was higher for mesh C (87.5%) compared with meshes A (62.5%) and B (41.6%). The severity of adhesions was also higher for mesh C (grade I in 14, II in 6, and III in 1) compared with mesh A (grade I in 10, II in 4, and III in 1 case) and B (all of them grade II). Histological inflammatory reaction was classified as mild in 23 cases of mesh A, 15 of mesh B, and 23 of mesh C. A moderate reaction was found in 1 case of mesh A, 4 cases of mesh B, and 1 case of mesh C. Severe reaction was induced in 5 cases of mesh B. Mesh B induced a higher inflammatory reaction compared with the other meshes. CONCLUSIONS: All meshes induced adhesions of different grades. Mesh B had fewer adhesions and more intense inflammation them did the others. PMID- 18402741 TI - Management of common bile duct stones: a ten-year experience at a tertiary care center. AB - BACKGROUND: The authors present their experience with the treatment of cholecystocholedocholithiasis. METHODS: From August 1995 to August 2005, we treated 3750 patients with symptomatic gallbladder stones, and 12.7% of these patients also had a choledocholithiasis. This percentage with reference to the literature is certainly due to intraoperative cholangiography (IOC) performed routinely. During the follow-up, the percentage of residual versus recurrent stones was evaluated. RESULTS: Only 2 patients (3.1%) of the laparoscopic choledochotomy and cholecystectomy group had retained stones discovered at 5 and 11 months after surgery. These patients received ERCP with good results. No patients had leaks or fistulas. Only one patient died from a massive pulmonary embolism (1.6%). CONCLUSION: We believe that the key to successful management of patients with CBD stones is a well-standardized institutional procedural algorithm consistent with the available local resources and technologies. PMID- 18402742 TI - Laparoscopic management of symptomatic Meckel's diverticula: a simple tangential stapler excision. AB - BACKGROUND: Meckel's diverticulum was first described about 400 years ago and continues to be a rare congenital disorder. Laparoscopic surgery for Meckel's diverticulum has been described in mostly case reports. We present our series of patients with symptomatic Meckel's diverticulum. METHODS: We have treated 12 patients with symptomatic Meckel's diverticulum from 1994 through 2006 at our institution. All the patients presented with features of either appendicitis or peritonitis, some with a vague abdominal mass. Clinical diagnosis of Meckel's diverticulum was made in only 4 patients. Diagnostic laparoscopy confirmed Meckel's diverticulitis in all patients. Laparoscopic stapler resection of the lesions was performed for all patients, tangential excision in 10 and wedge excision in 2. RESULTS: The incidence of Meckel's diverticulum at our institution is 0.3%. The majority of patients were male children. There were no staple-line leaks in any case. All patients recovered well postoperatively, and the day of discharge was in the range of the fourth to the seventh POD. Heterotopic gastric mucosa was found in the majority of the diverticula. Eight patients were followed up for 24 months, and 4 patients reported for follow-up after 45 months and were found to be symptom-free. DISCUSSION: The diagnosis of Meckel's diverticulitis is rarely made preoperatively. Surgical resection is indicated only if the diverticulum is symptomatic or if the base is narrow. Traditionally, open wedge resection (including the anterior wall of the ileum) of the diverticulum is the treatment. We think that a simple tangential stapler resection can also be performed, with good outcome. CONCLUSION: Laparoscopy is useful in both diagnosis and treatment. Laparoscopic resection of Meckel's diverticulum is feasible and ideal, especially when performed in specialized centers. PMID- 18402743 TI - Laparoscopic cholecystectomy can be safely performed in a resource-limited setting: the first 49 laparoscopic cholecystectomies in Yemen. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) is the gold standard for gallstone disease. Many studies have confirmed the safety and feasibility of LC and have shown that it is comparable regarding complications to open cholecystectomy (OC). The aim of this study was to evaluate the outcomes of LC including safety, feasibility in a resource-poor setting like Yemen, and also to compare the outcomes of LC with those of OC. METHODS: This was a prospective, nonrandomized, comparative study of 112 patients who were admitted to Alburaihy Hospital with a diagnosis of gallstone disease and underwent cholecystectomy from July 1998 to March 2004. Hospital stay, duration of operation, postoperative analgesia, and morbidity due to wound infection, bile leak, common bile duct (CBD) injury, missed CBD stone, bleeding, subphrenic abscess, and hernia were evaluated. Patients were followed up on an outpatient basis. RESULTS: Forty-nine patients underwent LC and 63 patients underwent OC. The mean age of LC patients was 43.96 years and of OC patients was 44.63 years. The 2 groups were similar in terms of age (p=0.740) and sex (p=0.535). No significant difference was found in the incidence of acute cholecystitis between the 2 groups (p=0.000). The mean operative duration for LC was 39.88 minutes versus 56.76 minutes for OC (p=0.000), and the mean hospital stay was 1.63 and 5.38 days for LC and OC, respectively (p=0.000). A drain was used frequently in OC (p=0.000). LC patients needed less analgesia (p=0.000). The morbidity rate in LC was 12.2% versus 6.3% for OC, which was not statistically significant (p=0.394), (p>0.05). Wound infection and bile leak were more common with LC. No mortalities were reported in either group. CONCLUSION: An experienced surgeon can perform LC safely and successfully in a resource-limited setting. As in other studies, LC outcomes were better than OC outcomes. PMID- 18402744 TI - Evaluation of efficacy of four laparoscopic needle drivers. AB - INTRODUCTION: To evaluate the impact of needle driver design on laparoscopic suturing skills by experts and novices. METHODS: Three experienced laparoscopic surgeons and 3 novice junior residents were asked to perform a fixed set of suturing tasks in a laparoscopic pelvic-trainer. The laparoscopic needle drivers compared were (1) the Ethicon driver (E 705R), (2) Karl Storz (KS) pistol grip (26173 KC), (3) KS finger grip (26167 SK), and (4) KS palm grip (26173 ML). Times were recorded for each operator to grasp and position a needle for suturing in a particular angle, as well as to throw a horizontal and a vertical stitch and tie a single square knot using 2-0 Vicryl suture with a taper CT-1 needle. Subsequently, participants were asked to complete a subjective questionnaire rating the drivers. RESULTS: The average suturing time provided the most discriminatory power in comparing the needle drivers. For experienced operators, the KS pistol grip allowed faster suturing times than did the KS finger grip and the KS palm grip but not the Ethicon driver. For novice users, the Ethicon driver allowed faster suturing times than did the KS finger grip but not the KS pistol grip or the KS palm grip. In the subjective questionnaire, the KS pistol grip received the highest scores, and the KS finger grip received the lowest scores. CONCLUSION: Novice laparoscopists performed best with the KS pistol grip as well as the Ethicon laparoscopic needle drivers while experienced laparoscopists performed best with the pistol grip KS needle driver. PMID- 18402745 TI - Reactionary hemorrhage in gynecological surgery. AB - BACKGROUND: Bleeding is a major complication in contemporary gynecological surgery. We discusses this rare, albeit potentially serious, unexpected complication. The authors mean by "reactionary," hemorrhage that occurs within the first 24 hours after surgery. More or less, all gynecological surgeons have had to deal with this situation at some stage of their career. The seriousness of this complication stems from the fact that often the surgeon is not in the immediate vicinity to promptly step in and treat the patient. Nevertheless, the key to successful management is prompt diagnosis, immediate resuscitation, and operative intervention. METHODS: By using the collective hospital database, we reviewed 719 patient records. The authors operated on these patients between November 1990 and March 2007 (inclusive) in one hospital, Royal Brisbane and Women's Hospital, the main teaching hospital in Brisbane, Queensland, Australia. The procedures performed in the 719 patients comprised 476 vaginal hysterectomies and 243 laparoscopic hysterectomies. Both public and private patients were included. The objective of the review was to establish the incidence of postoperative reactionary hemorrhage following the initial operation, as defined by the number of patients returning to the operating theatre (OT) because of postoperative hemorrhage within 24 hours of the initial hysterectomy. RESULTS: Of the 719 patients, 6 experienced reactionary postoperative hemorrhage, 3 each in the vaginal hysterectomy and laparoscopic hysterectomy groups. That would make the incidence of postoperative reactionary hemorrhage 0.6% in the vaginal hysterectomy and 1.2% in the laparoscopic hysterectomy group. None of these 6 patients had any preoperative hemorrhagica diatheses. There was neither ureteric, bladder, intestinal, nor any other injuries in the whole series. No long-term complications or mortalities occurred. CONCLUSION: Reactionary postoperative hemorrhage is a rare, albeit serious, complication of contemporary gynecological surgery; this complication may occur despite meticulous surgical technique. The key to successful management is prompt diagnosis, urgent resuscitation, and return to the OT to arrest the bleeding. PMID- 18402746 TI - Intestinal obstruction by capsule endoscopy in a patient with radiation enteritis. AB - BACKGROUND: Video capsule endoscopy is in widespread use as a diagnostic modality. Although capsule endoscopy is generally considered safe, several prior reports have documented capsules' failure to progress through narrowed areas of intestine. Symptomatic retention and obstruction by capsule endoscopy have not yet been reported in the setting of radiation enteritis. METHODS: We report a case of a patient with a history of pelvic radiation who underwent capsule endoscopy to identify an occult intestinal bleeding source after conventional modalities were not diagnostic. RESULTS: The patient was noted to have capsule retention several days longer than was expected, and video images of the distal bowel showed edema, narrowing, and ulceration consistent with radiation enteritis. The patient developed a symptomatic bowel obstruction requiring resection of this segment of bowel, including the impacted capsule. CONCLUSION: A history of abdominal or pelvic irradiation in patients with occult gastrointestinal bleeding should serve as a relative contraindication to video capsule endoscopy. The risk of obstruction and possible need for surgical intervention should be clearly outlined for such patients if they are to undergo this diagnostic maneuver. PMID- 18402747 TI - Endoscopic stenting of colovaginal fistula: the transanal and transvaginal "kissing" wire technique. AB - Interventional endoscopy is a rapidly evolving field allowing surgeons and endoscopists to approach surgical conditions nonoperatively. Stenting of benign colorectal disease has been limited due to technical issues and lack of long-term data. Colovaginal fistula can be a challenging condition to treat. In this report, we describe the technical aspect and results of endoscopic stenting of benign colovaginal fistula in 2 patients, using the combined transanal and transvaginal approach. PMID- 18402748 TI - Laparoscopic colotomy repair following colonoscopic polypectomy. AB - BACKGROUND AND OBJECTIVES: The unfortunate complication of a colotomy resulting from a colonoscopic polypectomy can be disastrous. Using the versatility of laparoscopic surgery, we aim to provide a treatment algorithm for these colonoscopic perforations. METHODS: We report a case of cecal perforation, in a 70 year old female, following colonoscopic polypectomy that was treated successfully with laparoscopic application of an Endo-GIA linear stapler. RESULTS: Four months following staple resection of her cecal perforation, the patient is asymptomatic and has undergone a repeat colonoscopy without any sequelae. CONCLUSIONS: Laparoscopic techniques can be safely applied in the treatment of colonic perforations following therapeutic polypectomies. A suggested management algorithm is provided highlighting the role laparoscopy may provide in selected patients. PMID- 18402749 TI - The perforated intrauterine device: endoscopic retrieval. AB - BACKGROUND: Uterine perforation is the most serious complication associated with an intrauterine contraceptive device (IUD). Minimally invasive techniques, such as hysteroscopy and advanced laparoscopy, are ideally suited to the diagnosis and surgical management of the perforated IUD. CASE REPORTS: Three cases of uterine perforation caused by an IUD and treated with endoscopic surgery are presented. In all 3 cases, the IUD was located by using x-rays, ultrasonography, or pelvic magnetic resonance imaging. Diagnostic laparoscopy was performed to identify the specific location of the IUD and to remove it. All patients recovered without incident. CONCLUSION: The gynecologic surgeon should acquire familiarity with the complications of, and proficiency in managing, perforated and ectopic IUDs by using modern surgical techniques that permit the patient's rapid return to health. PMID- 18402750 TI - Pregnant noncommunicating rudimentary uterine horn with placenta percreta. AB - BACKGROUND: To report a placenta percreta in a 7-week gestational rudimentary noncommunicating uterine horn pregnancy. METHODS: A 28-year-old woman with no complaints presented with a rudimentary uterine horn pregnancy at 7-weeks gestation. The diagnosis was suspected by ultrasonography and diagnosed by laparoscopy. Laparoscopic excision of the rudimentary uterine horn and ipsilateral salpingectomy were performed, as well as biopsy of several peritoneal endometriosis lesions. RESULTS: A 7-week gestation pregnancy with placenta percreta was identified in the rudimentary uterine horn. No communication was found with the right unicornuate uterus. Endometriosis was confirmed. Clinical outcome was favorable. CONCLUSION: Placenta percreta may occur in rudimentary uterine horn pregnancies, but accidents may be avoided by an early diagnosis and surgical management. However, in young women who desire pregnancy, planned laparoscopic resection of a rudimentary uterine horn revealed accidentally should be discussed. PMID- 18402751 TI - Laparoscopic transhiatal treatment of large epiphrenic esophageal diverticulum. AB - BACKGROUND: Epiphrenic diverticulum is an uncommon disorder of the distal third of the esophagus. We report the case of a 73-year-old woman with a large symptomatic esophageal epiphrenic diverticulum, diffuse nonspecific esophageal dysmotility, and a hiatal hernia. METHODS: Surgery was indicated by the patient's symptoms, the size of the diverticulum (maximum diameter 10 cm), and the associated esophageal motor disorder. Preoperative study included barium swallow, upper gastrointestinal endoscopy, and esophageal manometry. A laparoscopic transhiatal diverticulectomy associated with a Heller myotomy, hiatoplasty, and a Dor's fundoplication was carried out. The overall operative time was 230 minutes. RESULTS: No intraoperative complications occurred. Gastrografin swallow performed on postoperative day 4 did not show any signs of leakage from the staple line. The postoperative hospital stay was 5 days. The patient was readmitted 10 days after discharge complaining of fever and chest pain. A new Gastrografin swallow demonstrated a small leak from the staple line successfully treated with 3 weeks of total enteral nutrition. CONCLUSION: The laparoscopic approach to epiphrenic diverticulum is feasible. Postoperative Gastrografin swallow is not 100% sensitive in detecting small suture-line leaks if a preexisting esophageal motility disorder is present. In case of late postoperative fever and pleural effusion, a suture-line leak should be suspected. Conservative management of the small suture-line leak should be considered as an effective therapeutic option. PMID- 18402752 TI - Twenty years of laparoscopic cholecystectomy: Philippe Mouret--March 17, 1987. PMID- 18402753 TI - Expression of mRNAs encoding oestrogen receptor (ER) alpha and ERbeta, androgen receptor and progesterone receptor during gonadal and follicular development in the marsupial brushtail possum (Trichosurus vulpecula). AB - The objective of the present study was to determine which ovarian cells express mRNAs for oestrogen (ERalpha and ERbeta), androgen (AR) and progesterone (PR) receptors during ovarian and follicular development in the brushtail possum. Expression of ERalpha and/or ERbeta mRNA was observed from birth, initially in cells of the blastema, then in the medullary cords from Day 20. ERalpha was expressed in the oocytes and granulosa cells of secondary and antral follicles. Preovulatory follicles did not express ERalpha mRNA, although their oocytes were not examined for any gene. ERbeta mRNA was observed in oocytes at all follicular stages examined, but was not consistently observed in granulosa or theca cells. Expression of AR mRNA before Day 40 was very faint; thereafter, expression was observed in the medullary cords, peaking between Days 60 and 120. Oocytes, granulosa cells and theca of secondary and antral, but not preovulatory, follicles expressed AR mRNA. PR mRNA was expressed throughout the gonad by Day 20. Granulosa cells of some secondary and antral follicles and theca of antral follicles expressed PR mRNA. Thus, the expression of mRNAs encoding steroidogenic receptors in a time- and cell-specific manner supports a role for steroids in the process of ovarian follicular formation and growth. PMID- 18402754 TI - Oestrogen and progesterone receptor binding capacity and oestrogen receptor alpha expression (ERalpha mRNA) along the cervix of cycling ewes. AB - The aim of the present work was to study the oestrogen receptor (ER) and progesterone receptor (PR) binding capacity and the oestrogen receptor alpha (ERalpha) mRNA concentration in cranial and caudal cervix during the ovine oestrous cycle. Cervical samples of synchronised Corriedale ewes were obtained on Day 1 (n=7), 6 (n=6) or 13 (n=7) after oestrus detection (Day 0). The ER and PR binding capacity by ligand-binding assay and the ERalpha mRNA concentration by solution hybridisation in both cranial and caudal zones of the cervix were determined. The ER and PR binding capacity were higher (P<0.005) on Day 1 than on Days 6 and 13 in both cranial and caudal zones. The ERalpha mRNA concentrations were higher (P<0.0001) on Day 1 than on Days 6 and 13 only in the caudal zone. The PR binding capacity and ERalpha mRNA concentration were higher (P<0.005) in the caudal than in the cranial zone on Day 1. The ER and PR expression in the ovine cervix varied during the oestrous cycle in agreement with the known upregulation exerted by oestrogen and downregulation exerted by progesterone. Differences in ER and PR expression along the longitudinal axis of the ovine cervix were found, reflecting histological and functional differences between the cranial and caudal zones. PMID- 18402755 TI - Homeodomain protein HLX is expressed primarily in cytotrophoblast cell types in the early pregnancy human placenta. AB - Homeobox genes are a large family of transcription factors. Of these, the HLX homeobox gene (previously known as HLX1 and HB24) is important for normal placentation. We have previously shown that HLX mRNA expression is significantly reduced in fetal growth-restricted human placentae compared with control placentae. In this study, a rabbit polyclonal antibody to the homeodomain protein HLX was raised and characterised. Western analysis revealed a protein of 50 kDa. HLX protein was detected in cellular nuclei in the cytotrophoblast-derived cell lines HTR8/SVneo, SGHPL-4, JEG-3, JAR and BeWo. Dual labelling with cytokeratin 7 was used to determine the spatial distribution of HLX in the early placenta and fetal membranes, showing both a perinuclear and punctate nuclear distribution for HLX. In the early pregnancy placenta HLX was localised to villous cytotrophoblast, and extravillous cytotrophoblast nuclei in the proximal regions of the cytotrophoblast cell columns, but was not detected at significant levels in the syncytiotrophoblast. In first trimester placental bed biopsies, HLX expression was not localised to the nucleus but instead was found in the cytoplasm. We conclude that HLX is primarily expressed in cytotrophoblast cell types in the human placenta and propose that HLX is involved in cytotrophoblast proliferation and downregulation of cell differentiation. PMID- 18402756 TI - Effect of increasing progesterone concentration from Day 3 of pregnancy on subsequent embryo survival and development in beef heifers. AB - Higher systemic progesterone in the immediate post-conception period is associated with an increase in embryonic growth rate, interferon-tau production and pregnancy rate in cattle. The objective of this study was to examine the effect of increasing progesterone concentration on Day 3 on subsequent embryo survival and development. Oestrus (Day 0) was synchronised in beef-cross heifers (n=210) and approximately two-thirds of the heifers were inseminated with semen from a proven sire, while the remainder were not inseminated. In order to produce animals with divergent progesterone concentrations, half of the animals received a progesterone-releasing intravaginal device (PRID) on Day 3 of the oestrous cycle, which was left in situ until slaughter. The four treatment groups were: (i) pregnant, high progesterone; (ii) pregnant, normal progesterone; (iii) non pregnant, high progesterone; and (iv) non-pregnant, normal progesterone. Animals were blood-sampled twice daily from Days 0 to 8 and once daily thereafter until slaughter on Days 5, 7, 13 or 16, corresponding to the 16-cell stage, the blastocyst stage, the beginning of elongation and the day of maternal recognition of pregnancy, respectively. Embryos were recovered by flushing the tract with phosphate-buffered saline and characterised by stage of development and, in the case of Days 13 and 16, measured. Data were analysed by mixed models ANOVA, Chi square analysis and Student's t-test where appropriate. Insertion of a PRID on Day 3 increased (P<0.05) progesterone concentrations from Day 3.5 onwards. There was no difference between treatments in the proportion of embryos at the expected stage of development on Days 5 or 7 (P>0.05). While not significantly different, the proportion of viable embryos recovered was numerically greater in the high progesterone group on both Day 13 (58 v. 43%) and Day 16 (90 v. 50%). Elevation of progesterone significantly increased embryonic length on Day 13 (2.24+/-0.51 mm v. 1.15+/-0.16 mm, P=0.034) and Day 16 (14.06+/-1.18 cm v. 5.97+/-1.18 cm, P=0.012). In conclusion, insertion of a PRID on Day 3 of the oestrous cycle increased serum progesterone concentrations on subsequent days, which, while having no phenotypic effect on embryonic development on Days 5 or 7, was associated with an increase in embryonic size on Days 13 and 16. PMID- 18402757 TI - Miniature ponies: 1. Follicular, luteal and endometrial dynamics during the oestrous cycle. AB - Follicular dynamics were studied during 12 interovulatory intervals (IOIs) and 36 preovulatory periods in Miniature mares. The percentage of IOIs with the following follicle events was: ovulatory wave with only one follicle>or=10 mm (55%), diameter deviation similar to previous reports in larger mares (25%) and minor waves emerging before or after the ovulatory wave (55%). Follicle data were compared among Miniature ponies, large ponies and Breton horses (n=12 IOIs per breed). The IOI was longer (P<0.001) in Miniature ponies (23.3+/-0.9 days) and in large ponies (23.9+/-0.5 days) than in Breton horses (20.3+/-0.7 days). The Miniature ponies had fewer (P<0.0001) growing follicles>or=10 mm per ovulatory wave (1.5+/-0.3) and more (P<0.0004) ovulatory waves (6/11) with only one follicle>or=10 mm than large ponies (9.8+/-0.8 and 0/12) and horses (5.8+/-0.9 and 0/12). Maximum diameter of the preovulatory follicle was smaller (P<0.003) in the Miniature ponies (38.3+/-0.7 mm) than in the horses (44.5+/-1.4 mm), but the difference between breeds was slight (6%) compared with the difference in bodyweight (65%). Considering the small number of follicles per ovulatory wave, Miniature mares are a potential model for comparative studies in folliculogenesis within and among species. PMID- 18402758 TI - Miniature ponies: 2. Endocrinology of the oestrous cycle. AB - Plasma concentrations of FSH, LH, oestradiol and progesterone were studied daily during 12 interovulatory intervals and 21 periovulatory periods in nine Miniature ponies. The peak of the FSH surge that was temporally associated with emergence of the future ovulatory follicle occurred when the follicle was approximately 9 mm, compared with a reported diameter of 13 mm in larger breeds. The ovulatory LH surge involved a slow increase between Days 13 and 18 (ovulation=Day 0; 0.6+/-0.1 ng day(-1)), a minimal increase or a plateau on Days 18 to 21 (0.04+/-0.1 ng day( 1)), and a rapid increase after Day 21 (2.2+/-0.4 ng day(-1); P<0.0001). The end of the plateau and the beginning of the rapid increase occurred on the day of maximum concentration in the oestradiol preovulatory surge. An unexpected mean increase and decrease in LH occurred (P<0.04) on Days 5 to 9. Concentrations of oestradiol and progesterone seemed similar to reported results in larger breeds. Results indicated that in Miniature ponies the peak of the FSH surge associated with emergence of the future ovulatory follicle occurred at a smaller diameter of the future ovulatory follicle than in larger breeds, the ovulatory LH surge increased in three phases, and the ovulatory LH surge was followed by an LH increase and decrease during the early luteal phase. PMID- 18402759 TI - Testosterone secretion, testicular histology and the cryopreservation of cauda epididymidal spermatozoa in the common ringtail possum (Pseudocheirus peregrinus). AB - The present study reports novel aspects of the reproductive biology of the male common ringtail possum (Pseudocheirus peregrinus). Plasma testosterone was measured through a stimulation test using the gonadotrophin-releasing hormone agonist, buserelin. Following intra-muscular administration of buserelin, there was an increase (P<0.05) in testosterone concentration in the peripheral circulation 4 h later. Quantitative testicular histology of this species was described for the first time. Eight stages of the seminiferous epithelium cycle were identified in 10 possums and their relative frequency determined. Spermatozoa were recovered from the cauda epididymides of hemi-castrated possums and cryopreservation conducted in straws (6 degrees C min(-1)) using final glycerol concentrations ranging between 2 and 20% in Tris-citrate egg yolk extender (v/v). Frozen straws were thawed and post-thaw motility, rate of motility, the percentage of live-dead spermatozoa and the percentage of sperm with swollen decondensed nuclei recorded. Similar to other marsupial sperm, common ringtail possum cauda epididymidal spermatozoa required high levels of glycerol (10-16%) in order to maintain post-thaw viability. PMID- 18402760 TI - Spermatozoa from a marsupial, the brushtail possum, contain beta1,4 galactosyltransferase. AB - Beta1,4-galactosyltransferase-I (GalTase-I) is one of the key molecules on the sperm surface of eutherian mammals that is likely to be involved in binding to the egg coat, the zona pellucida, to mediate sperm-egg interaction. In laboratory mice, the species for which most data are available, this protein functions as a receptor for the zona pellucida protein ZP3 of the oocyte and, upon binding, triggers the sperm acrosome reaction. In the present study, we investigated the presence and abundance of GalTase-I in epididymal sperm extracts of a marsupial, the brushtail possum, Trichosurus vulpecula. For this, spermatozoa were collected from cauda epididymides and the amount of beta1,4-galactosyltransferase activity in washed sperm extracts was compared with that of porcine spermatozoa. Overall beta1,4-galactosyltransferase enzyme activity was found to be more abundant in possum sperm extracts than those from porcine spermatozoa (P<0.05). Immunoblots with an antibody to mouse GalTase-I revealed that the molecular weight of possum spermatozoa GalTase-I was 66 kDa, which is similar to the molecular weight of GalTase-I in spermatozoa from eutherian mammals. The molecular weight of GalTase I was the same in sperm extracts collected from the caput and cauda epididymides. These results demonstrate that GalTase-I is indeed present in possum spermatozoa and thus it may be a gamete receptor molecule on the sperm surface of marsupials as well as those of eutherian mammals. PMID- 18402761 TI - Theoretical investigation into the dissolved oxygen levels in follicular fluid of the developing human follicle using mathematical modelling. AB - Oxygen levels in the follicle are likely to be critical to follicle development. However, a quantitative description of oxygen levels in the follicle is lacking. Mathematical modelling was used to predict the dissolved oxygen levels in the follicular fluid of the developing human follicle. The model predictions showed that follicular fluid dissolved oxygen levels are highly variable among follicles, due to the unique geometry of individual follicles. More generally, predictions showed that oxygen levels in follicular fluid increase rapidly during the initial early antral stages of follicle growth before peaking in the later early antral phase. Follicular fluid dissolved oxygen levels then decline through to the beginning of the pre-ovulatory phase, from which they increase through to ovulation. Based on the best available parameter estimates, the model predictions suggest that the mean dissolved oxygen levels in human follicular fluid during the late antral and pre-ovulatory phases range between 11 and 51 mmHg (approximately 1.5-6.7 vol%). These predictions suggest that the human ovarian follicle is a low-oxygen environment that is often challenged by hypoxia, and are in agreement with only some published data on follicular fluid oxygen levels. Predictions are discussed in relation to follicle health and oocyte culture. PMID- 18402762 TI - Measurement of rest and activity in newborn lambs using actigraphy: studies in term and preterm lambs. AB - The present study used actigraphy to monitor rest-activity cycles in lambs. We employed an Actiwatch Activity Monitor, which was secured on the lamb's neck in 13 term lambs and six preterm lambs. Activity measurements began on the day of delivery and lasted for 7.3+/-0.7 days. All lambs exhibited bouts of activity, lasting from approximately 2 to 60 min, separated by periods of inactivity of about equal duration. There was a progressive increase in the frequency and intensity of activity bouts with age, and a decrease in duration. In relation to postnatal age, preterm lambs had a significantly lower frequency and intensity of activity bouts compared with term lambs and significantly longer mean active bout duration. However, in relation to post-conceptual age, preterm animals were less active at birth, but thereafter the trajectory for activity development was steeper compared with the term lambs. These differences between term and preterm lambs may be due to several factors including differences in: (1) the lengths of time the two groups spent in utero and as neonates as a proportion of the perinatal period, which could influence the rate of muscle and bone growth; (2) prenatal and postnatal hormonal profiles; and (3) maternal care. We also found differences in postnatal motility in male and female lambs, with the trajectory of activity increasing in males at Days 4-5, which could be due, in part at least, to sex differences in both prenatal and postnatal hormonal profiles. PMID- 18402763 TI - Relationships of basal metabolic rate, relative testis size and cycle length of spermatogenesis in shrews (Mammalia, Soricidae). AB - The aim of the present study was to determinate the cycle length of spermatogenesis in three species of shrew, Suncus murinus, Sorex coronatus and Sorex minutus, and to assess the relative influence of variation in basal metabolic rate (BMR) and mating system (level of sperm competition) on the observed rate of spermatogenesis, including data of shrew species studied before (Sorex araneus, Crocidura russula and Neomys fodiens). The dynamics of sperm production were determined by tracing 5-bromodeoxyuridine in the DNA of germ cells. As a continuous scaling of mating systems is not evident, the level of sperm competition was evaluated by the significantly correlated relative testis size (RTS). The cycle durations estimated by linear regression were 14.3 days (RTS 0.3%) in Suncus murinus, 9.0 days (RTS 0.5%) in Sorex coronatus and 8.5 days (RTS 2.8%) in Sorex minutus. In regression and multiple regression analyses including all six studied species of shrew, cycle length was significantly correlated with BMR (r2=0.73) and RTS (r2=0.77). Sperm competition as an ultimate factor obviously leads to a reduction in the time of spermatogenesis in order to increase sperm production. BMR may act in the same way, independently or as a proximate factor, revealed by the covariation, but other factors (related to testes size and thus to mating system) may also be involved. PMID- 18402764 TI - Embryo-endometrial expression of leukemia inhibitory factor in the golden hamster (Mesocricetus auratus): increased expression during proestrous and window of implantation stages. AB - Leukemia inhibitory factor (LIF) is a pleiotropic IL-6 family cytokine and its maternal uterine expression is critical for mouse blastocyst implantation. In the golden hamster (Mesocricetus auratus), although the blastocyst hatching phenomenon is quite interesting and LIF is shown to regulate hatching, information is not available on the embryonic and uterine expression of LIF and hormonal regulation of LIF expression during the peri-implantation period. The present investigation is aimed at studying embryonic and uterine expression of LIF during preimplantation hamster development. We observed embryonic expression of LIF mRNA and protein in the 8-cell, morula and blastocyst stages. In cycling females, uterine LIF mRNA expression was maximal during the oestrogen-dominant phase of the oestrous cycle, i.e. proestrous stage. Interestingly, during pregnancy, both LIF mRNA and protein were highly upregulated on Days 3.5 and 4 ('window of implantation'), implying a role for this cytokine in blastocyst hatching and implantation. Cell type-specific localisation of LIF mRNA and protein was observed predominantly in luminal epithelium and uterine glands with faint staining being detected in the stroma. The hamster uterus encoded a approximately 4.2 kb LIF transcript whose coding region, when cloned and sequenced, showed a high degree of identity to the murine cDNA counterpart. These data demonstrate that: (1) hamster preimplantation embryos show LIF mRNA and protein expression; (2) uterine expression of LIF mRNA and protein was dependent on elevated levels of circulating oestrogen, and (3) there is a possible functional association of LIF with the peri-implantation development in the golden hamster. PMID- 18402765 TI - Membrane interactions of G proteins and other related proteins. AB - Guanine nucleotide-binding proteins, G proteins, propagate incoming messages from receptors to effector proteins. They switch from an inactive to active state by exchanging a GDP molecule for GTP, and they return to the inactive form by hydrolyzing GTP to GDP. Small monomeric G proteins, such as Ras, are involved in controlling cell proliferation, differentiation and apoptosis, and they interact with membranes through isoprenyl moieties, fatty acyl moieties, and electrostatic interactions. This protein-lipid binding facilitates productive encounters of Ras and Raf proteins in defined membrane regions, so that signals can subsequently proceed through MEK and ERK kinases, which constitute the canonical MAP kinase signaling cassette. On the other hand, heterotrimeric G proteins undergo co/post translational modifications in the alpha (myristic and/or palmitic acid) and the gamma (farnesol or geranylgeraniol) subunits. These modifications not only assist the G protein to localize to the membrane but they also help distribute the heterotrimer (Galphabetagamma) and the subunits generated upon activation (Galpha and Gbetagamma) to appropriate membrane microdomains. These proteins transduce messages from ubiquitous serpentine receptors, which control important functions such as taste, vision, blood pressure, body weight, cell proliferation, mood, etc. Moreover, the exchange of GDP by GTP is triggered by nucleotide exchange factors. Membrane receptors that activate G proteins can be considered as such, but other cytosolic, membranal or amphitropic proteins can accelerate the rate of G protein exchange or even activate this process in the absence of receptor mediated activation. These and other protein-protein interactions of G proteins with other signaling proteins are regulated by their lipid preferences. Thus, G protein-lipid interactions control the features of messages and cell physiology. PMID- 18402766 TI - Site-specific modification of positively-charged surfaces on human serum albumin by malondialdehyde. AB - Malondialdehyde (MDA), a lipid peroxidation product, reacts with lysine residues in proteins. Human serum albumin (HSA) is a major target of MDA-modification of serum proteins. To identify, the modification sites of HSA by MDA in vitro, MDA treated HSA was digested with a protease and the resulting peptides were subjected to liquid chromatography-tandem mass spectrometry. We identified six peptides, which contained a N-propenal adduct at Lys136, Lys174, Lys240, Lys281, Lys525, and Lys541, and revealed that Lys525 is the most reactive residue for MDA modification. Analysis of electrostatic surface potential of a 3-D model structure of HSA indicates that Lys525 is located at the center of positively charged grooves. The results of this study indicate that the modification of proteins by lipid-derived aldehydes may be influenced by the electrostatic potential of the protein surface. PMID- 18402767 TI - A newly identified dependence receptor UNC5H4 is induced during DNA damage mediated apoptosis and transcriptional target of tumor suppressor p53. AB - UNC5H4 is a netrin-1 receptor UNC5H family member. In this study, we found that UNC5H4 is a direct transcriptional target of p53. During adriamycin (ADR) mediated apoptosis, UNC5H4 was significantly induced in p53-proficient U2OS cells but not in p53-deficient H1299 cells. Enforced expression of p53 induced UNC5H4. Consistent with these results, siRNA-mediated knockdown of p53 in U2OS cells attenuated ADR-dependent induction of UNC5H4. Indeed, we found four putative p53 responsive elements within intron 1 of UNC5H4 gene. Luciferase reporter assay and ChIP analysis demonstrated that, among them, two tandem elements respond to exogenous p53 which is efficiently recruited onto them. Furthermore, enforced expression of UNC5H4 remarkably reduced number of drug-resistant colonies in p53 proficient cells but not in p53-deficient cells, suggesting that UNC5H4-induced apoptosis is dependent on p53 status. siRNA-mediated knockdown of UNC5H4 rendered U2OS cells resistant to ADR. Collectively, our present results suggest that UNC5H4 amplifies p53-dependent apoptotic response. PMID- 18402768 TI - Simultaneous knockdown of BRAF and expression of INK4A in melanoma cells leads to potent growth inhibition and apoptosis. AB - Abnormal BRAF and p16INK4A co-exist in 60% of melanomas. BRAF mutation also occurs in 80% of benign nevi where it turns-on p16INK4A resulting in proliferative senescence; loss of p16INK4A removes the inhibitory block leading to melanoma development. Since only melanomas with wild-type BRAF have amplified CDK4 and cyclin D1 genes, p16INK4A-CDK4/6-cyclin D pathway is viewed as linearly downstream of BRAF. Thus, co-occurrence of aberrant BRAF and INK4A may be remnant of changes during melanoma formation without functional significance. To explore this notion, we simultaneously knocked down BRAF (via siRNA) and expressed INK4A cDNA in melanoma cells and observed enhanced growth inhibition. Notably, although each alone had no statistically significant effect on apoptosis, co-expression of BRAF siRNA and INK4A cDNA caused potent apoptosis, which was associated with up regulation of BIM and down-regulation of BCL2. Our results suggest that aberrant BRAF and INK4A cooperate to promote proliferation and survival of melanoma cells. PMID- 18402769 TI - Rational design of minimal hypoxia-inducible enhancers. AB - The hypoxia-inducible factor (HIF) activates transcription via binding to the highly variable hypoxia-responsive elements (HREs). All hypoxia-inducible constructs described to date utilize multimers of naturally occurring HREs. Here, we describe the rational design of minimal hypoxia-inducible enhancers, conceptually equivalent to using an optimized HIF-binding site (HBS) as the building block. Optimizations of the HBS, spacing between HBSs, the distance from the minimal promoter, and orientation of HBSs allowed us to design constructs with high hypoxic activity. Activation of the 4xopt HBS (36bp) construct by hypoxia or HIF-1alpha and HIF-2alpha was comparable with that of the 4xEPO HRE (208bp) construct. The strong synergism between the properly arranged optimized HBSs was due to stimulation of high affinity HIF binding. Our data prove, for the first time, that it is possible to assemble artificial hypoxia-inducible enhancers from a single type of regulatory element-optimized HBS. PMID- 18402770 TI - S-Adenosyl-l-methionine activates the cardiac ryanodine receptor. AB - S-Adenosyl-l-methionine (SAM) is the biological methyl-group donor for the enzymatic methylation of numerous substrates including proteins. SAM has been reported to activate smooth muscle derived ryanodine receptor calcium release channels. Therefore, we examined the effects of SAM on the cardiac isoform of the ryanodine receptor (RyR2). SAM increased cardiac sarcoplasmic reticulum [(3)H]ryanodine binding in a concentration-dependent manner by increasing the affinity of RyR2 for ryanodine. Activation occurred at physiologically relevant concentrations. SAM, which contains an adenosine moiety, enhanced ryanodine binding in the absence but not in the presence of an ATP analogue. S-Adenosyl-l homocysteine (SAH) is the product of the loss of the methyl-group from SAM and inhibits methylation reactions. SAH did not activate RyR2 but did inhibit SAM induced RyR2 activation. SAH did not alter adenine nucleotide activation of RyR2. These data suggest SAM activates RyR2 via a site that interacts with, but is distinct from, the adenine nucleotide binding site. PMID- 18402771 TI - Essential roles of caspases and their upstream regulators in rotenone-induced apoptosis. AB - In the present study, we examined whether caspases and their upstream regulators are involved in rotenone-induced cytotoxicity. Rotenone significantly inhibited the proliferation of oral cancer cell lines in a dose-dependent manner compared to normal oral mucosal fibroblasts. Flow cytometric analysis of DNA content showed that rotenone treatment induced apoptosis following G2/M arrest. Western blotting showed activation of both the caspase-8 and caspase-9 pathways, which differed from previous studies conducted in other cell types. Furthermore, p53 protein and its downstream pro-apoptotic target, Bax, were induced in SAS cells after treatment with rotenone. Rotenone-induced apoptosis was inhibited by antioxidants (glutathione, N-acetylcysteine, and tiron). In conclusion, our results demonstrate significant involvement of caspases and their upstream regulators in rotenone-induced cytotoxicity. PMID- 18402772 TI - Up-regulation of the cellular level of Escherichia coli PTS components by stabilizing reduced transcripts of the genes in response to the low oxygen level. AB - When Escherichia coli cells were grown with limited levels of oxygen, the glucose induced transcription of ptsG was decreased whereas deletion of the arcA gene partially restored it, which was consistent with the previous report that the ArcA protein represses ptsG transcription. However, under this circumstance, we found that the level of EIICB(Glc) protein encoded by the ptsG gene was rather increased. This paradoxical phenomenon can be explained by the delayed turnover of ptsG mRNA in cells anaerobically grown in the presence of glucose. Finally, our data showed that anaerobic expression of the ptsHIcrr operon is also enhanced by increasing the longevity of the reduced mRNA levels. PMID- 18402773 TI - CMTM3 can affect the transcription activity of androgen receptor and inhibit the expression level of PSA in LNCaP cells. AB - CMTM is a novel family of proteins linking chemokines and TM4SF. Several members of this family are highly expressed in testes and regulate androgen receptor (AR) transcription activity. One member of this family, CMTM3, has the highest expression level in testes and contains one leucine zipper and two LXXLL motifs. As assessed with the dual-luciferase reporter assay, overexpression of CMTM3 represses AR transactivation, while knocking down it can increase AR transactivation. Moreover, CMTM3 inhibits prostate-specific antigen (PSA) expression in LNCaP cells at both mRNA and protein levels with no obvious influence on AR expression. Taken together, CMTM3 may play some roles in the maturation and maintenance of the male reproduction. PMID- 18402774 TI - C-RAF activation promotes BAD poly-ubiquitylation and turn-over by the proteasome. AB - BAD, a member of the BCL2 family, exhibits an original mode of regulation by phosphorylation. In the present report, we examine the role of the kinase C-RAF in this process. We show that the inducible activation of C-RAF promotes the rapid phosphorylation of BAD on Serine-112 (Ser-75 in the human protein), through a cascade involving the kinases MEK and RSK. Our findings reveal a new aspect of the regulation of BAD protein and its control by the RAF pathway: we find that C RAF activation promotes BAD poly-ubiquitylation in a phosphorylation-dependent fashion, and increases the turn-over of this protein through proteasomal degradation. PMID- 18402775 TI - FTY720 inhibits S1P-mediated endothelial healing: relationship to S1P1-receptor surface expression. AB - The phosphorylated derivative of the immunosuppressant FTY720 interacts with and modulates the function of sphingosine-1-phosphate (S1P)-receptors. We observed a significant reduction of endothelial surface binding of a S1P(1)-specific antibody after FTY720 treatment of 6h and longer, which was associated with a reduced healing after mechanic injury, impaired angiogenesis and enhanced adhesion molecule expression. FTY720 (5h) had no impact on the expression of S1P(1)- or S1P(3)-encoding transcripts. Notably, pre-treatment of cells with FTY720 for only 30min, which did not reduce S1P(1) surface expression, inhibited the rapid S1P- and SEW2871- (a S1P(1) agonist) induced cortical actin formation and cell migration. FTY720 was effective at concentrations as low as 5nM. FTY720 at therapeutic concentrations may be harmful by impairing important endothelial functions. Interestingly, FTY720 inhibited endothelial actin remodelling and cell migration without decreasing S1P(1) surface expression. PMID- 18402776 TI - Bmx regulates LPS-induced IL-6 and VEGF production via mRNA stability in rheumatoid synovial fibroblasts. AB - Discordant cytokine production is characteristic of chronic inflammatory conditions like rheumatoid arthritis (RA), and anti-cytokine therapeutics are becoming routinely used to treat RA in the clinic. Fibroblasts from rheumatoid synovium have been shown to contribute to cytokine production in inflamed joints; likewise these cells also produce cytokines in response to inflammatory mediators signalling through Toll like receptors (TLRs). Tyrosine kinase activity is essential to LPS-induced cytokine production, and we have previously implicated a role for the Tec kinase, Bmx, in inflammatory cytokine production. Here we show that Bmx kinase activity in RASF is increased following LPS stimulation and that Bmx is involved in the regulation of LPS-induced IL-6 and VEGF production via mRNA stabilisation. This is an important insight into the regulation of VEGF, which is involved in a wide range of different pathologies, and may lead to more effective design of novel anti-inflammatory/angiogenic therapeutics for conditions such as RA. PMID- 18402778 TI - Characterization of minor bands of STR amplification reaction of FVIII gene by PCR cloning. PMID- 18402779 TI - Study of the plasmatic levels of tryptophan and kynurenine throughout pregnancy. PMID- 18402780 TI - Development of high-performance magnetic chemiluminescence enzyme immunoassay for alpha-fetoprotein (AFP) in human serum. AB - BACKGROUND: A high-performance chemiluminescence enzyme immunoassay (CLEIA) for alpha-fetoprotein (AFP), a tumor marker for the diagnosis of hepatocellular carcinoma (HCC), was constructed by using magnetic particles (MPs) as both the immobilization matrix and separation tools. METHODS: A double sandwiched immunocomplex was formed through the reaction among anti-fluorescein isothiocyanate (FITC) antibody coated MPs, FITC-labeled anti-AFP antibody, AFP antigen, and alkaline phosphatase (ALP)-labeled anti-AFP antibody. The subsequent chemiluminescence reaction of ALP with 4-methoxy-4-(3-phosphate-phenyl)-spiro (1,2-dioxetane-3,2'-adamantane) (AMPPD) gave light intensity that was directly proportional to the amount of analyte present in the samples. The effects of several physicochemical parameters, including the concentration of FITC-labeled anti-AFP antibody, the dilution ratio of ALP-labeled anti-AFP antibody, the volume of MPs and substrate, the immunoreaction time and other relevant variables upon the immunoassay were studied and optimized. RIA and microplate CLEIA were used as comparison methods. RESULTS: The proposed method had a sensitivity of 3.0 ng/ml, low cross reactivities, and an assay time of 1 h. The linear range was 0 1200 ng/ml through using MPs and is useful for samples with extremely high AFP concentrations without dilution while avoiding the hook effect. The intra- and inter-assay precision was <3% and <5%. The present method has been successfully applied to the detection of AFP human serum with recoveries from 90 to 108%, and showed a good correlation with the commercially available AFP RIA kit. CONCLUSIONS: This proposed assay provided apparent advantages over microplate CLEIA and RIA, and facilitated the development of high-throughput screening and automated operation systems in the clinical practice. PMID- 18402781 TI - Serum brain-derived neurotrophic factor in patients with type 2 diabetes mellitus: Relationship to glucose metabolism and biomarkers of insulin resistance. AB - OBJECTIVES: The aims of this study were to measure serum levels of brain-derived neurotrophic factor (BDNF) in patients with type 2 diabetes mellitus (T2DM) and to investigate the association of these BDNF levels with biomarkers of glucose metabolism and insulin resistance. DESIGN AND METHODS: We studied 112 patients with T2DM and 80 age- and gender-matched control subjects. RESULTS: Serum BDNF levels were significantly lower in patients with T2DM compared to control subjects (15.5+/-5.2 ng/mL vs. 20.0+/-7.3 ng/mL, P<0.01). In patients with T2DM, BDNF levels were significantly higher in females than in males (P<0.01). In the female patients, BDNF was positively related to immunoreactive insulin (IRI) (rho=0.458, P<0.05) and HOMA-R (rho=0.444, P<0.05). Stepwise multiple regression analysis showed a significant relationship between BDNF and IRI (F=5.294, P<0.05) in female patients with diabetes. CONCLUSIONS: These findings suggest that BDNF may contribute to glucose metabolism. PMID- 18402782 TI - Kinetics of hydrogen peroxide elimination by astrocytes and C6 glioma cells analysis based on a mathematical model. AB - Oxidative stress is implicated in a variety of disorders including neurodegenerative diseases, and H(2)O(2) is important in the generation of reactive oxygen and oxidative stress. In this study, we have examined the rate of extracellular H(2)O(2) elimination and relevant enzyme activities in cultured astrocytes and C6 glioma cells and have analyzed the results based on a mathematical model. As compared with other types of cultured cells, astrocytes showed higher activity of glutathione peroxidase (GPx) but lower activities for GSH recycling. C6 cells showed relatively low GPx activity, and treatment of C6 cells with dibutyryl-cAMP, which induces astrocytic differentiation, increased catalase activity and H(2)O(2) permeation rate but exerted little effect on other enzyme activities. A mathematical model [N. Makino, K. Sasaki, N. Hashida, Y. Sakakura, A metabolic model describing the H(2)O(2) elimination by mammalian cells including H(2)O(2) permeation through cytoplasmic and peroxisomal membranes: comparison with experimental data, Biochim. Biophys. Acta 1673 (2004) 149-159.], which includes relevant enzymes and H(2)O(2) permeation through membranes, was found to be fitted well to the H(2)O(2) concentration dependences of removal reaction with the permeation rate constants as variable parameters. As compared with PC12 cells as a culture model for neuron, H(2)O(2) removal activity of astrocytes was considerably higher at physiological H(2)O(2) concentrations. The details of the mathematical model are presented in Appendix. PMID- 18402783 TI - The proteome of the human neuroblastoma cell line SH-SY5Y: an enlarged proteome. AB - The human neuroblastoma cell line SH-SY5Y (ATCC: CRL-2266) is widely used as a neural cellular model system. The hitherto existing proteome data (115 proteins) are here extended. A total of 1103 unique proteins of this cell line were identified using 2D-LC combined with MALDI-TOF/TOF-MS, SDS-PAGE with nano-LC MS/MS, N-terminal COFRADIC analysis with nano-LC-MS/MS and 2D-PAGE with MALDI TOF/TOF-MS peptide mass fingerprinting. The obtained proteome profile of this cell line is discussed. PMID- 18402784 TI - Interaction of the major protein from bovine seminal plasma, PDC-109 with phospholipid membranes and soluble ligands investigated by fluorescence approaches. AB - The major protein from bovine seminal plasma, PDC-109 binds selectively to choline phospholipids on the sperm plasma membrane and plays a crucial role in priming spermatozoa for fertilization. The microenvironment and accessibility of tryptophans of PDC-109 in the native state, in the presence of phosphorylcholine (PrC) and phospholipid membranes as well as upon denaturation have been investigated by fluorescence approaches. Quenching of the protein intrinsic fluorescence by different quenchers decreased in the order: acrylamide>succinimide>>Cs(+)>I(-). Ligand binding afforded considerable protection from quenching, with shielding efficiencies following the order: dimyristoylphosphatidylcholine (DMPC)>lysophosphatidylcholine (Lyso-PC)>PrC. This has been attributed to a partial penetration of the protein into the DMPC membranes and Lyso-PC micelles, as well as a further stabilization of the binding due to the interaction of PDC-109 with lipid acyl chains and the resulting tightening of the protein structure, leading to a decreased accessibility of the tryptophan residues. Red-edge excitation shift (REES) studies yielded REES values of 4 nm for both native and denatured PDC-109, whereas reduced and denatured protein gave a REES of only 0.5 nm, clearly indicating that the structural and dynamic features of the microenvironment around the tryptophan residues are retained even after denaturation, presumably due to the constraints imposed on the protein structure by disulfide bonds. Upon binding of PDC-109 to DMPC membranes and Lyso-PC micelles the REES values were reduced to 2.5 and 1.0 nm, respectively, which could be due to the penetration of some parts of the protein, especially the segment containing Trp-90 into the membrane interior, where the red-edge effects are considerably reduced. PMID- 18402785 TI - Anomalous origin of additional coronary artery arising from the pulmonary artery in complex congenital heart disease. AB - Anomalous origin of coronary arteries is a rare congenital defect and usually presents as an isolated defect. In only 5% of cases, it may be associated with other cardiac anomalies. In this article, we present a case of a newborn with a very complex congenital cardiac disease accompanied with an anomalous origin of an additional coronary artery. PMID- 18402786 TI - A 4-year-old girl with giant left atrial diverticulum resulting in severe mitral regurgitation. AB - Left atrial diverticulum is rare in a pediatric patient. A 4-year-old girl underwent a left atrial plication concomitant with mitral valve replacement, and early postoperative course was uneventful. This patient has remained asymptomatic with no evidence of recurrent diverticular formation after 9 months of follow-up. PMID- 18402787 TI - Truffle's revenge: a pig-eating fungus. AB - A 77-year-old female initially presented with symptomatic mitral valve stenosis involving a bioprosthesis that had been implanted 8 months earlier for myxomatous mitral valve disease and severe valvular regurgitation. The patient was taken for a second mitral valve replacement due to stenosis. Intraoperatively, the bioprosthetic mitral valve was noted to have an unusual clot-like mass on the atrial side. Initial fungal smears were positive for yeast stains, and pathology revealed extensive colonization by thick filamentous fungus with apparent true hyphae, pseudohyphae, and yeast forms. The fungus was identified as Hormographiella aspergillata, the asexual form of Coprinus cinereus, a common inky cap mushroom that grows in the lawn. She was treated with 6 weeks of liposomal amphotericin B and then switched to voriconazole for long-term (lifelong) suppressive therapy in the setting of a new mechanical mitral valve. The only other reported case of infective endocarditis caused by a Coprinus species occurred in a 53-year-old man who had developed native aortic valve fungal endocarditis and died [J Med Microbiol (1971);4(3):370-4]. The valve isolate was identified as probable C. cinereus. PMID- 18402788 TI - Late coronary thrombosis following brachytherapy for vasospastic angina. PMID- 18402789 TI - Primary atrial fibrosarcoma of the heart. AB - The primary fibrosarcoma of the heart is a rare tumor. There are no exact numbers about its incidence, but even among malignant cardiac tumors, only around 3% are fibrosarcomata. Symptoms are nonspecific with signs of right or left heart failure depending on localization. Diagnosis is thereby often delayed. Primary treatment of choice is surgery, followed by several possible postsurgical adjuvant strategies. Survival is poor with a mean of about 11 months. PMID- 18402790 TI - Pathology of isolated atrial infarction: case report and review of the literature. AB - Infarction of the cardiac atria occurs more frequently than is commonly considered. Ischemic damage to the atrial myocardium is usually associated with infarction of cardiac ventricles, but isolated infarction of an atrium can occur and may be of clinical significance. We present an unusual case of an isolated right atrial infarct with an infarction-related endocardial thrombus that mimicked a mass lesion near the inferior vena cava. The presentation and causes of isolated atrial infarction are discussed. PMID- 18402791 TI - Angiogenesis, tumor necrosis factor-alpha and procoagulant factors in coronary artery giant aneurysm of a fatal infantile Kawasaki disease. AB - BACKGROUND: Kawasaki disease (KD) is an infantile febrile illness of unknown origin characterized by clinical, laboratory and histopathologic features of systemic vasculitis. METHODS AND RESULTS: We report a 3-month-old female infant with incomplete KD who suddenly died despite intravenous immunoglobulin, aspirin, steroid and heparin treatment. Postmortem examination confirmed the echocardiographically detected giant coronary aneurysms and showed occlusive thrombosis in the giant aneurysm of the left anterior descending coronary artery, associated with neoangiogenesis, macrophage infiltration and immunostaining for tissue factor (a strong initiator of the coagulation cascade), thrombopoietin receptor and tumour necrosis factor-alpha. CONCLUSIONS: These findings show the association of angiogenesis, tumor necrosis factor-alpha and procoagulant factors, with macrophage infiltration in coronary artery aneurysms of a fatal infantile KD. PMID- 18402792 TI - Total anomalous origin of the coronary circulation from the right pulmonary artery. AB - Origin of both coronary arteries from a branch of the pulmonary artery is rare and has not been reported as a cause of sudden unexpected death. We report autopsy findings of a 14-day male infant in previously good health who died suddenly. From the proximal right pulmonary artery arose a single coronary artery, which branched into the right and left main coronary arteries. The right proximal coronary artery coursed between the aorta and pulmonary trunk, and the left main traveled along the anterior proximal aorta. The distal course of the coronary circulation was normal. There was concentric subendocardial healing myocardial infarction. No other congenital anomalies were identified. Total anomalous origin of the coronary arteries from the pulmonary trunk or artery should be considered in cases of sudden unexpected death in infants. PMID- 18402793 TI - Cannabis and cocaine: a lethal cocktail triggering coronary sudden death. AB - The case of a 31-year-old man who died suddenly, with a history of drug abuse and no risk factors for coronary artery disease, is herein reported. Postmortem examination disclosed critical stenosis due to a fibrocellular atherosclerotic plaque complicated by occlusive thrombosis at the level of the left anterior descending coronary artery. Chemico-toxicological analysis revealed high levels of cocaine, tetrahydrocannabinol, and metabolites. When dealing with an acute coronary syndrome in young people, with or without a history of drug abuse, toxicological examination should always be performed to rule out unnatural causes of myocardial ischemia. PMID- 18402794 TI - Giant right coronary artery aneurysm presenting as a paracardiac mass. AB - Coronary artery aneurysms, commonly atherosclerotic, are usually asymptomatic and can be diagnosed incidentally during investigation for ischemic heart disease or on autopsy. An asymptomatic giant right coronary artery aneurysm presenting as a paracardiac mass discovered on autopsy is reported here. Even though there was great concern due to its large size, complex atherosclerotic lesions, and potential obstruction of blood flow, it had no direct link to the cause of death. We also review the literature on giant right coronary artery aneurysms exceeding 5 cm in the last 10 years. PMID- 18402795 TI - Surgical pathology of 104 tricuspid valves (2000-2005) with classic right-sided Ebstein's malformation. AB - BACKGROUND: Ebstein's anomaly has been described extensively in autopsy material. However, there have been no large surgical pathology series of this malformation. OBJECTIVE: To review clinical and surgical pathologic features of a large number of cases of Ebstein's anomaly from a single institution. METHODS: Review of medical histories, surgical reports, and surgical pathology reports at the Mayo Clinic (2000-2005). RESULTS: Among 104 patients, the mean age was 31 years (2 months-79 years), and 57% were female. Common ECG abnormalities included right bundle branch block (58%), first-degree heart block (31%), preexcitation (18%), and nonspecific intraventricular conduction delay/block (15%). Moreover, 74% had inter-atrial communication, 13% mitral valve prolapse, and 5% bicuspid aortic valve. Clinically, all had tricuspid regurgitation (severe in 74%), and 17% of anterior leaflets were fenestrated. No tricuspid valve was calcified. Surgically, tricuspid tissue was removed during replacement in 99% and repair in 1%. The anterior tricuspid leaflet was resected in 98%, and its length was 0.81-9.3 cm/m2 body surface area (mean, 3.3). Characteristically, leaflets were large and had irregular shapes and numerous short cordal or direct myocardial insertions. One tricuspid valve had two papillary fibroelastomas. None had clinical or pathologic evidence of active or healed endocarditis. CONCLUSIONS: Among patients with Ebstein's malformation, tricuspid valve tissue almost exclusively was removed during valve replacement and represented the anterior leaflet. Valve tissue was generally large, irregularly shaped, and associated with insertion of short cords or myocardial stumps. Interestingly, although appreciably deformed, Ebstein valves were not associated with infective endocarditis. PMID- 18402796 TI - Severe dilated cardiomyopathy as a consequence of Ecstasy intake. AB - Dilated cardiomyopathy (DCM) is one of the most common causes of heart failure with a prevalence of 1:2500. There are several primary and secondary etiologic factors, including gene mutations, infection agents, particularly viruses, toxins, autoimmune, and systemic disorders, and pheochromocytoma, neuromuscular, metabolic, mitochondrial, and nutritional disorders. However, a precise diagnosis can be reached only in no more than 50% of all cases. Herein, we report a rare case of hepatic damage and severe DCM as a consequence of relatively popular socially used narcotic-Ecstasy (3,4-methylenedioxy-N-methylamphetamine [MDMA]). PMID- 18402797 TI - Myocardial sympathetic innervation in patients with impaired glucose tolerance: relationship to subclinical inflammation. AB - AIM: This study was designed to assess cardiac adrenergic nerve activity, using iodine (I)-123-labeled metaiodobenzylguanidine (MIBG), in patients with impaired glucose tolerance (IGT) and to investigate its relation to circulating levels of proinflammatory cytokines. METHODS: We studied 22 patients with IGT (aged 34-68 years) and 18 age-matched healthy controls, using I-123 MIBG cardiac imaging. The early (10 min) and late (4 h) heart to mediastinum MIBG uptake (H/M) ratio and washout were calculated. Levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-a), and its soluble receptor [soluble TNF receptor II (sTNFRII)] were measured by immunoassay of blood samples from patients and controls. RESULTS: The early and late MIBG uptake was lower (both P<.001) and the WR was higher (P<.001) in patients than in controls. The analysis showed innervation defects in 20 of the 22 patients. Nearly half (45.4%) showed severe adrenergic innervation defects in both the inferior wall and the apex. Regarding cytokines, patients showed significantly elevated TNF-a (P=.005), sTNFRII (P<.001), and IL-6 (P<.001) levels compared to controls. IL-6 and sTNFRII were found to correlate with the WR (r=0.468, P=.028 and r=0.455, P=.034, respectively). CONCLUSION: Patients with IGT show reduced MIBG cardiac uptake with a segmental pattern. The reduced cardiac sympathetic innervation was related to the elevated proinflammatory cytokine levels and could be considered an index of early atherosclerotic process in these patients. PMID- 18402798 TI - Dedifferentiation of atrial cardiomyocytes in cardiac valve disease: unrelated to atrial fibrillation. AB - BACKGROUND: Valvular heart disease has become an important public health concern. The increased wall stress and underlying disease entity associated with mitral valve disease provide unfavorable circumstances for atrial cardiomyocytes. The expression of the alpha-smooth muscle actin isoform is considered characteristic of cardiomyocyte dedifferentiation (embryonic cardiomyocyte), and cardiomyocyte dedifferentiation may indicate an adaptive state, enabling cardiomyocytes to survive despite unfavorable circumstances. METHODS: This study comprised 20 adult patients with symptomatic severe mitral valve disease and moderate to severe tricuspid valve disease and without coronary artery disease undergoing valve operations for congestive heart failure. Ten patients had persistent atrial fibrillation and 10 patients had never been in atrial fibrillation by history and electrocardiograms before surgery. Atrial tissues of the right atrial appendage were obtained during surgery. RESULTS: Immunohistochemical study demonstrated that alpha-smooth muscle actin protein expression was not altered by atrial fibrillation, and alpha-smooth muscle actin protein expression in atrial tissues was higher in patients with sinus rhythm than in those with atrial fibrillation (the percentage of cells that were alpha-smooth muscle actin-positive was 51.5+/ 34.9% for right atria from patients in sinus rhythm vs. 16.2+/-15.0% for right atria from patients with atrial fibrillation) (P<.03). Semiquantitation of alpha smooth muscle actin by immunoblotting of extracts from atrial tissues showed similar findings as in the immunohistochemical observations: that is, atrial fibrillation did not influence the expression of alpha-smooth muscle actin protein. Interstitial fibrosis represented 43.2+/-13.9% of the right atrial tissue in the sinus group, whereas interstitial fibrosis comprised 49.8+/-8.2% of the right atrial tissue in the atrial fibrillation group (P=.320). CONCLUSIONS: Dedifferentiation of atrial cardiomyocytes occurs in patients with cardiac valve disease, even without atrial fibrillation. PMID- 18402799 TI - Histopathology of the cardiac conduction system in sudden intrauterine unexplained death. AB - BACKGROUND: Sudden intrauterine unexplained death (SIUD) is one of the most heartbreaking tragedies that any parent can experience. It remains poorly understood and incompletely examined both morphologically and functionally. The aim of this work is to examine the likely role of cardiac conduction system in relation to sudden and unexplained fetal death. METHODS: We analyzed and compared the autopsy results in 15 cases of SIUD (6 males and 9 females, ranging in age from 35 to 40 weeks) and 11 cases of intrauterine explained death (IED). A complete autopsy was performed, focusing on the examination of the cardiac conduction system on serial sections. RESULTS: The following findings were observed: resorptive degeneration (33% of SIUD, 36% of IED), dispersion or septation of the atrioventricular (AV) junction (60% of SIUD, 64% of IED), islands of the conduction system in the central fibrous body (80% of SIUD, 73% of IED), Mahaim fibers (20% of SIUD), cartilaginous metahyperplasia (20% of SIUD, 18% of IED), an AV node (AVN) tongue (13% of SIUD), hemorrhage of the cardiac conduction system (7% of SIUD, 9% of IED), left-sided bifurcation (7% of SIUD), an intramural right bundle (7% of SIUD), central fibrous body hypoplasia (7% of SIUD), and thickening of the conduction system arteries (13% of SIUD). CONCLUSIONS: Most of the abnormal cardiac conduction findings were detected only in SIUD and were absent in controls, i.e., Mahaim fibers, AVN tongue, left-sided bifurcation, intramural right bundle, and central fibrous body hypoplasia. We are convinced that these cardiac conduction abnormalities, in association with altered neurovegetative stimuli, could underlie potentially malignant arrhythmias. PMID- 18402800 TI - Discrepancies in initial death certificate diagnoses in sudden unexpected out-of hospital deaths: the role of cardiovascular autopsy. AB - BACKGROUND: The accuracy of death certificates issued for out-of-hospital sudden deaths has been questioned. METHODS: We retrospectively studied a series of consecutive autopsies performed at two community hospitals. RESULTS: Fifty-four autopsies in which the deaths were sudden and that occurred outside the hospital were retained for study. The indication for autopsy was largely driven by the wishes of family or physician who was uncertain about the diagnosis. The overall discrepancy rate was 52%. The death certificate diagnosis, rendered before autopsy, was coronary artery disease in 44/54 autopsies (81%). At autopsy, coronary artery disease was the cause of death in 26 cases (48%), cardiomyopathy in 10 (18%), ruptured aneurysm in 8 (15%), pulmonary embolism in 7 (13%), and valve disease in 3 (6%). The diagnosis of coronary artery disease on the death certificate was accurate only 50% of the time. The discrepancy rate was lowest in patients with a history of cardiovascular disease (33%) and was 60% in patients with no prior medical history. The accuracy rate of death certificates was under one third in cases of cardiomyopathy, valve disease, ruptured aneurysm, pulmonary embolism, and valve disease. CONCLUSIONS: We conclude that in a highly selected group of sudden deaths, in which there was often a question about cause of death, the rate of initial death certificate accuracy is only one half. Furthermore, coronary artery disease as the cause of death is less than 50%, far less than initial death certificate diagnoses would indicate. PMID- 18402801 TI - Induction of cardiovascular pathology in a novel model of low-grade chronic inflammation. AB - OBJECTIVE: Epidemiological and clinical evidence indicate that inflammatory processes play a pivotal role in a number of conditions associated with aging, including osteoporosis and cardiovascular diseases. The purpose of this study was to evaluate cardiovascular pathology and select inflammatory mediators of interest in a model of low-grade inflammation-induced osteopenia. METHODS: Slow release pellets were subcutaneously implanted in male rats to deliver 0, 3.3, or 33.3 microg of lipopolysaccharide (LPS)/day for 90 days. Tail blood was collected at 1, 2, and 3 months for differential white cell counts, and at the end of the study, hearts were harvested for histological and immunohistochemical evaluation. RESULTS: The low-grade inflammatory response was characterized by elevated peripheral blood neutrophils and monocytes. Histological examination of heart cross sections revealed increased fibrous tissue, infiltration of lymphocytes, accumulation of mast cells, and roughened intimal borders within the arteries and arterioles, consistent with vascular disease. Inflammatory mediators (cyclooxygenase-2, tumor necrosis factor-alpha, and interleukin-1 beta) were up regulated, and increased expression of platelet endothelial cell adhesion molecule-1 and receptor activator for NF-kappaB ligand was localized to the microvasculature endothelium. CONCLUSIONS: These findings suggest that inflammation induced by chronic exposure to LPS produces cardiovascular pathology in the smaller intramural arteries and arterioles and support the utility of this model for further mechanistic and therapeutic studies focused on the role of chronic inflammation in cardiovascular disease. PMID- 18402802 TI - Quadricuspid aortic valve with ascending aortic aneurysm: report of a case and discussion of embryological mechanisms. AB - Congenital quadricuspid aortic valves (QAVs) are rare phenomena. They are often associated with aortic insufficiency and significant morbidity. A QAV with an associated ascending aortic aneurysm is extremely rare, and these conditions might have a shared embryonic etiology. We describe the case of a patient with a QAV associated with aortic aneurysm. Pertinent literature on the QAV is reviewed, and embryological factors that may contribute to its pathogenesis are discussed. PMID- 18402803 TI - Pathology of inverted Takotsubo cardiomyopathy. AB - Myocardial dysfunction without coronary involvement may occur in acute cerebral diseases. The inverted Takotsubo pattern has been recently recognized as a novel heart neurologic stress-related syndrome. We report on the case of a 40-year-old woman presenting with massive subarachnoid hemorrhage and brain death. Echocardiography revealed an extensive left ventricular circumferential akinesis except at the apex. Histologic analysis of the heart confirmed the absence of myocardial infarction and revealed only sparse foci of myocyte necrosis with contraction bands in the akinetic areas. PMID- 18402804 TI - Primary cardiac cytotoxic T-cell lymphoma presenting with neurological deficits: a case report. AB - BACKGROUND: Primary cardiac lymphoma is extremely rare in immunocompetent patients. Clinical manifestations vary, and, most often, diagnosis is not made until autopsy. The majority of reported primary cardiac lymphoma cases have been of B-cell origin, while T-cell cardiac lymphomas have been extremely rare. Occasionally, lymphomas and other systemic malignancies clinically present as paraneoplastic neurological syndromes. METHODS: We report a unique case of primary cardiac peripheral T-cell lymphoma of cytotoxic phenotype, clinically presenting with neurological features of external ophthalmoplegia and lower cranial nerve paresis mimicking mitochondrial cytopathy, that was recognized at autopsy. Brain and thoracoabdominal viscera retrieved at autopsy were fixed in 10% buffered formalin and processed for paraffin embedding. In addition to routine histology, immunohistochemistry for immunophenotypic characterization of lymphoma cells was performed. Fresh skeletal muscle was processed for cryosectioning and histochemical staining. RESULTS: On gross examination, the heart showed multiple circumscribed, whitish nodules on both sides. Histological examination of these nodules revealed lymphomatous deposits-cells expressing CD45, CD2, CD3, CD5, CD7, CD8, perforin, and granzyme B. Histological sections from the brain showed foci of demyelination and patchy perivascular lymphoid cell aggregates in leptomeninges and within the parenchyma. These lymphoid cells expressed CD2, CD3, and CD5, with the T cells being predominantly CD4 (CD4:CD8>2), which was unlike the CD8-predominant lymphomatous infiltrate in the heart. Hence, these lymphoid cells in the brain, rather than disseminated lymphoma cells, were considered to be related to the demyelinating process. There was no evidence of lymphomatous deposits in the rest of the viscera examined. CONCLUSION: A diagnosis of primary cardiac peripheral T-cell lymphoma of cytotoxic phenotype clinically manifesting as paraneoplastic demyelinating lesions in the brain was described. PMID- 18402805 TI - Phytate inhibits bovine pericardium calcification in vitro. AB - OBJECTIVE: The present study examined the inhibitory effects of pyrophosphate, etidronate, and phytate on bovine pericardium calcification in vitro. METHODS: Bovine pericardium was glutaraldehyde fixed and then placed in a flow chamber in the presence of a synthetic physiological fluid alone (control) or the fluid plus various concentrations of pyrophosphate, etidronate, or phytate. Following a 96-h incubation, fragments were removed and assayed for calcification by measuring calcium and phosphorus levels. RESULTS: The data indicated that both pyrophosphate and etidronate at 1 mg/l (5.75 and 4.95 microM, respectively) inhibited bovine pericardium calcification, whereas neither agent had an effect at 0.5 mg/l (2.87 and 2.47 microM, respectively). Phytate was the most potent inhibitor of calcification, and the effects of this agent were apparent at levels as low as 0.25 mg/l (0.39 microM). CONCLUSIONS: While pyrophosphate, etidronate, and phytate were all able to inhibit bovine pericardium calcification in vitro, phytate was found to be the most effective. PMID- 18402806 TI - Multiple hydatid cystectomy of the heart necessitating LIMA to LAD anastomosis in a young patient. AB - Cardiac hydatid disease is very rare, even in endemic regions. Clinical manifestations included chest pain, anaphylactic shock, constrictive pericarditis, congestive heart failure, and arterial embolism. Surgery is the exclusive therapy, where the cysts are excised during open-heart surgery. The surgical approach therefore must be performed carefully, given the potential complications that surgery may bring. Because of the risk of potentially lethal complications, early diagnosis and definitive treatment are important. A 32-year old male patient was admitted with chest pain, weight loss, lethargy, and dizziness. On the transesophageal echocardiography study, a cystic mass (2.5 x 3 x 4.5 cm in dimension adjacent to the left ventricular posterior wall) that was divided into two by a septum was noted. Diagnosis of hydatidosis was confirmed with serologic tests (ELISA and indirect immunofluorescence). Echinococcosis, also known as hydatid disease, is common in several regions of the world, for example, the Mediterranean countries, the Middle East, South America, and East Africa. While performing pericystectomy in the anterior left ventricular wall, we noticed that there were three cysts, contrary to the preoperative diagnosis pointing a single one, and it was impossible to effectively complete the procedure without compromising anterosuperiorly displaced left anterior descending artery (LAD). We decided to go on bypass, arrest the heart, and complete the pericystectomy at the cost of injuring LAD and grafting the left internal mammary artery to LAD. Microscopic examination of the cyst showed a germinal layer and an avascular, eosinophilic, chitinous layer that confirmed the diagnosis of hydatid cyst. The patient was discharged on the fifth postoperative day on albendazole medication. PMID- 18402807 TI - C-reactive protein induces high-mobility group box-1 protein release through activation of p38MAPK in macrophage RAW264.7 cells. AB - BACKGROUND: C-reactive protein (CRP) is widely used as a sensitive biomarker for inflammation. Increasing evidence suggests that CRP plays a role in inflammation. High-mobility group box-1 (HMGB1), a primarily nuclear protein, is passively released into the extracellular milieu by necrotic or damaged cells and is actively secreted by monocytes/macrophages. Extracellular HMGB1 as a potent inflammatory mediator has stimulated immense curiosity in the field of inflammation research. However, the molecular dialogue implicated between CRP and HMGB1 in delayed inflammatory processes remains to be explored. METHODS AND RESULTS: The levels of HMGB1 in culture supernatants were determined by Western blot analysis and enzyme-linked immunosorbent assay in macrophage RAW264.7 cells. Purified CRP induced the release of HMGB1 in a dose- and time-dependent fashion. Immunofluorescence analysis revealed nuclear translocation of HMGB1 in response to CRP. The binding of CRP to the Fc gamma receptor in RAW264.7 cells was confirmed by fluorescence-activated cell sorter analysis. Pretreatment of cells with IgG-Fc fragment, but not IgG-Fab fragment, efficiently blocked this binding. CRP triggered the activation of p38MAPK and ERK1/2, but not Jun N-terminal kinase. Moreover, both p38MAPK inhibitor SB203580 and small interfering RNA significantly suppressed the release of HMGB1, but not the MEK1/2 inhibitor U 0126. CONCLUSION: We demonstrated for the first time that CRP, a prominent risk marker for inflammation including atherosclerosis, could induce the active release of HMGB1 by RAW264.7 cells through Fc gamma receptor/p38MAPK signaling pathways, thus implying that CRP plays a crucial role in the induction, amplification, and prolongation of inflammatory processes, including atherosclerotic lesions. PMID- 18402808 TI - Early morphological changes of an Amplatzer Septal Occluder explanted at heart transplant. AB - A 59-year-old male with severe biventricular heart failure presented with worsening dyspnea and angina. Following left ventricular assist device insertion, an Amplatzer Septal Occluder (ASO) was required as the patient's patent foramen ovale reopened. Seven months later, the patient underwent heart transplantation. The excised ASO was covered with a thick layer of host tissue on both right and left atrial sides, and organized thrombus was found between the metal wires. The foreign materials, the Nitinol wire and the synthetic (Dacron) fabric, showed a reactive infiltrate of macrophages, multinucleate giant cells, and scattered mononuclear cells. PMID- 18402809 TI - Histological analysis of aortic dissections following previous cardiovascular surgery. AB - BACKGROUND: Aortic dissections have been reported as a rare complication post cardiovascular surgery due to areas of cross clamping, suture lines and cannulation sites as locations for peri-operative injury. This study aimed to review the histological features of aortic tissues of 11 patients with aortic dissections who underwent previous cardiovascular surgery and to identify evidence of trauma within the area of surgery. METHODS: A review of our records from January 2000 to July 2005 showed that 11 patients had developed a postoperative aortic dissection. Surgically excised aortic tissues were obtained either at surgery or at autopsy for further gross and histological analysis. Clinical records were also reviewed for relevant history. RESULTS: The mean age of the 11 patients in this study was 60.1+/-12.5 years. The year of index surgery for all patients was between 1960 and 2005, and the interval between index surgery and admission for re-operation was 13.8+/-12.9 years (range, 1 day to 45 years). Five patients (45%) demonstrated aortic intimal thickening, two of which showed evidence of atherosclerotic plaques. Three (27%) exhibited cystic medial change. All cases showed mild to moderate medial changes near or at the site of traumatic injury, as demonstrated by a sharp discontinuity in the media of the aorta. Other than the four aortic tissues obtained at autopsy, long-term follow up for the remaining patients was not obtained. CONCLUSION: Evidence of previous trauma in close proximity to cannulation, cross-clamping and/or aortotomy sites was found in all patients in this series. All deaths in this series were found in the acute group rather than in the chronic group. It is important that the occurrence of dissections in patients undergoing previous cardiac surgery be not only reported, but analyzed by both the clinician and the pathologist. Hopefully, these efforts may identify particular patient risk factors and clarify preventative measures peri-operatively which may decrease the incidence of these potentially lethal complication. PMID- 18402810 TI - Polytetrafluoroethylene-covered stents in the venous and arterial system: angiographic and pathologic findings in a swine model. AB - BACKGROUND: Although polytetrafluoroethylene (PTFE) is now used as a covering in a wide variety of vascular stents, little is known about the resultant pathologic and angiographic changes in the arterial and venous system after stent deployment. METHODS: Atrium PTFE-covered stents were deployed in the abdominal aorta and the inferior vena cava (IVC) of two Yorkshire pigs. Angiograms were performed after deployment and at recatheterization. A complete histopathological exam was performed after sacrifice, and angiographic and histology findings were then compared between the two types of blood vessels. RESULTS: The arterial stents demonstrated no stenosis on angiography and minimal in-stent intimal proliferation by histopathology. A uniform layer of neointima coated the inside of the stent by 1 month with no obvious thrombus formation. In contrast, the venous stents demonstrated angiographic evidence of significant in-stent stenosis of variable degrees and the histology showed increased intimal hyperplasia with fibrin deposition. PTFE from both the arterial and venous stents had variable amounts of tissue ingrowth with cells observed to be growing into and through the PTFE fibers. CONCLUSIONS: Atrium PTFE-covered stents have excellent short-term angiography and histopathology in the arterial system, while stents in the venous system developed rapid in-stent intimal hyperplasia. PTFE fibers from both types of vasculature permitted cells to grow through and within them. PMID- 18402811 TI - Human monocyte-derived hemangioma-like endothelial cells: evidence from an in vitro study. AB - BACKGROUND: Hemangiomas are highly prevalent in newborns and infants and can lead to severe complications. However, the pathogenesis of hemangiomas is still unknown. This study was designed to examine the potential of human monocytes to differentiate into hemangioma endothelial cells. METHODS: Purified monocytes from adult human peripheral blood were cultured under a conditional culture environment supplemented with basic fibroblast growth factor and vascular endothelial growth factor. Cells cultured for 2 weeks were subjected to histological and immunochemical examinations in order to determine the expression of specific markers for hemangioma endothelial cells. RESULTS: Monocytes cultured for 2 weeks in angiogenic medium expressed human erythrocyte-type glucose transporter protein, FcgammaRII, and several other endothelial markers, all of which are deemed specific markers for hemangioma endothelial cells. However, neither CD133 nor alpha smooth muscle actin was detected in our monocyte culture. CONCLUSION: Our data suggested that monocytes are capable of differentiating into hemangioma endothelial cells under the angiogenic stimulation from microenvironment of proliferative hemangioma. PMID- 18402812 TI - Neuronal ceroid lipofuscinosis in a 31-year-old woman presenting as biventricular heart failure with restrictive features. AB - A 31-year-old woman presented with dyspnea and left-sided chest discomfort and was found to have biventricular heart failure with impaired ventricular filling. Clinically, she was thought to have restrictive cardiomyopathy or constrictive pericarditis. Transmission electron microscopy of myocardial tissue unexpectedly revealed crosshatched, curvilinear, and fingerprint depositions, which were characteristic for neuronal ceroid lipofuscinosis. Cardiac involvement by this inherited disorder is discussed in light of the findings in this patient and in 15 other reported cases. PMID- 18402813 TI - Paraoxonase-1 deficiency in mice predisposes to vascular inflammation, oxidative stress, and thrombogenicity in the absence of hyperlipidemia. AB - BACKGROUND: Paraoxonase-1 is a polymorphic enzyme that is strongly associated with circulating high-density lipoproteins. The absence of paraoxonase-1 in mice has been shown to promote diet-induced atherosclerosis. As paraoxonase-1 has been recently shown to be a lactonase, its functional role remains to be fully elucidated. We explored additional vascular changes in Pon1 knockout mice in the absence of atherogenic diet challenge. METHODS: Early steps in atherogenesis, namely, leukocyte rolling and firm adhesion, were measured using intravital microscopy. Vascular oxidative status was determined by lucigenin-derived chemiluminescence. Arterial thrombosis was determined by in vivo carotid thrombosis assay. Gene expressions were determined by reverse transcription polymerase chain reaction. RESULTS: We observed a twofold increase in leukocyte adhesion, but no significant change in leukocyte rolling in Pon1(-/-) mice versus wild-type controls. This finding is correlated with a 1.6-fold increase in aortic mRNA levels of P-selectin (P<.016), a 1.3-fold up-regulation in Vcam1 (P=.096), and a 1.5-fold up-regulation in Icam1 (P=.016). Aortic Tnfalpha mRNA expression was unchanged. Pon1(-/-) mice were also found to show a threefold increase in aortic superoxide production rate (P=.04). Furthermore, carotid thrombosis assay revealed a 57% reduction in time to occlusion in Pon1(-/-) mice (P<.001). In spite of such vascular proinflammatory phenotypes, we observed no change in plasma levels of inflammatory cytokines or in hepatic mRNA expression of serum amyloid A. CONCLUSION: Our data revealed significant vascular changes in adhesion, oxidative stress, and thrombotic tendencies in Pon1(-/-) mice in the absence of hyperlipidemia and systemic inflammation. PMID- 18402814 TI - Development of atherosclerosis in Balb/c apolipoprotein E-deficient mice. AB - BACKGROUND: Since its creation in 1992 by gene inactivation via gene targeting, the apolipoprotein E "knockout" mouse has become the most widely used rodent model for the study of atherosclerosis. Commercially available apolipoprotein E( /-) mice are bred on a C57BL/6J background. The goal of the present study was to investigate the development of atherosclerosis in apolipoprotein E-deficient mice generated on a Balb/c background. METHODS: We compared serum cholesterol concentrations and the development of atherosclerotic lesions in heterozygous Balb/c [apolipoprotein E(+/-)] mice fed regular rodent chow, Balb/c apolipoprotein E-deficient mice fed regular chow, and Balb/c apolipoprotein E deficient mice fed a high-fat diet for up to 30 weeks. Expression of the chemokine JE (murine homologue of MCP-1), as well as the adhesion molecules E selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule 1, in the aortas of knockout mice fed a high-fat diet was measured by enzyme linked immunosorbent assay. RESULTS: Balb/c apolipoprotein E-deficient mice develop atherosclerotic lesions in a reproducible temporal and morphological pattern. Total serum cholesterol concentrations in Balb/c apolipoprotein E deficient mice fed regular chow or a high-fat diet, respectively, closely parallel those reported for C57BL/6J apolipoprotein E-deficient mice. The expression of all three adhesion molecules in the aorta follows a similar temporal pattern, peaking in the first 15 weeks, whereas JE concentrations peak around 23 weeks. CONCLUSION: The availability of Balb/c apolipoprotein E deficient mice will facilitate the study of atherosclerosis in a mouse strain that can concomitantly develop other pathological states that are not readily inducible in mice with the C57BL/6J background. PMID- 18402815 TI - Perivascular epithelioid cell tumor of the pericardium: a case report. AB - A large pericardial mass, which was found to be a perivascular epithelioid cell tumor, was excised from a 20-year-old man. To our knowledge, this is the first report of this distinctive neoplasm as a pericardial mass. PMID- 18402816 TI - Monocyte-platelet complexes on CD14/CD16 monocyte subsets: relationship with ApoA I levels. A preliminary study. AB - The adhesion of the monocytes to the endothelium and their extravasation into the intima are key steps in atherogenesis. Studies showed the essential role of L selectin (CD62-L), expressed by the monocytes, and the platelets by forming complexes with monocytes. The delipided apolipoprotein (Apo) A or high-density lipoprotein (HDL) has antiinflammatory effects on monocytes and can bind platelets (monocyte-platelet complexes [MPCs]). The aim of this study was to identify a possible relationship between the MPCs, the monocyte subset, and ApoA I/HDL serum levels in vivo. Platelet-monocyte complexes were estimated by flow cytometry in 16 volunteers. Monocyte-platelet interaction was characterized by the percentage of monocytes coexpressing the constitutive platelet marker, glycocalicin gpIb-alpha (CD42b; CD42b+monocytes in %, MPC%). Monocytes were divided into four subsets based on lipopolysaccharide receptor (CD14) and FcgammaIII receptor (CD16) expression (CD14++/CD16-, G1; CD14++/CD16+, G2; CD14+/CD16-, G3; and CD14+/CD16+, G4). HDL and ApoA-I levels were measured by routine laboratory techniques. MPC% in the different subsets were G1=8.1+/-3.4%, G2=21.2+/-14%, G3=18+/-12.6%, and G4=22.3+/-14.3% (analysis of variance: P<.001). MPC% in the entire monocyte population was negatively correlated to ApoA-I (R= 0.71, P=.001). The relationship between ApoA-I and MPC% was found mainly in the subsets G1 (R=-0.67, P=.001) and G2 (R=-0.61, P=.01). MPC% was not correlated with any other lipids or lipoprotein or high-sensitivity C-reactive protein. When whole blood was incubated with HDL/ApoA-I, no modification of platelet CD42b fluorescence was observed, indicating that there is no direct interaction between the HDL/ApoA-I and the CD42b fluorescence. Among the monocytes, the G2 subset appeared to have the highest extravasation potential. Indeed, we previously showed that those cells overexpressed CD62-L, and we observed in this work that they were coated with platelets more than the G1 cells. The G2 subset could be more directly involved in the development of atherosclerotic lesions. PMID- 18402818 TI - Pediatric heart tumors. AB - Heart tumors in children are rare and consist primarily of nonneoplastic hamartomas (rhabdomyoma and fibroma) that may spontaneously regress. Despite the lack of aggressive growth potential, these tumors may result in significant morbidity as well as mortality based on size and location in the heart. Other rare tumors, including histiocytoid cardiomyopathy, hemangioma, germ cell tumors, and sarcomas, may occur in children in the heart and are discussed. PMID- 18402817 TI - Primary cardiac undifferentiated sarcoma of the right atrium presenting with cardiac tamponade. AB - This report illustrates the serial imaging of a primary cardiac undifferentiated sarcoma of the right atrium using echocardiography, chest X-ray, and computed tomography. Transthoracic echocardiography on presentation showed an extensive mass of the right atrial free wall with an impending cardiac tamponade. Symptoms were controlled with pericardiocentesis, pericardial window, and radiotherapy but recurred 8 months later with pleural effusion and tumor spread to the great arteries. Primary cardiac sarcoma (PSC) is a rare and aggressive malignancy that is usually diagnosed late due to its nonspecific symptoms. Cytology and cardiac biopsy may be negative, and suspicion for the tumor is warranted in recurrent pericardial effusion. Analogous to parietal pleural biopsy in lung tumors with pleural effusion, parietal pericardial biopsy may be positive in PSC of the right atrium with pericardial effusion. Echocardiography is the major diagnostic tool and aids pericardiocentesis. Pericardial window may be useful for recurrent pericardial effusion but does not preclude its reaccumulation. There is no proven effective treatment for PSC, and treatments include surgical resection, cardiac transplant, chemotherapy, and radiotherapy. Despite its poor prognosis, symptomatic relief is important and attainable. PMID- 18402819 TI - Effect of niacin on adipocyte leptin in hypercholesterolemic rabbits. AB - BACKGROUND: Leptin may play an important role in the development of atherosclerosis. Several transcription genes [including peroxisome proliferator activated receptor gamma (PPARgamma) and CD36] involved in lipid and glucose metabolism and inflammatory processes may correlate to leptin expression. The aim of this study was to investigate the effect of niacin on serum leptin levels in hypercholesterolemic rabbits and the expression of leptin, PPARgamma, and CD36 in adipocytes from hypercholesterolemic rabbits. METHODS: Eighteen rabbits fed with high-cholesterol diet for 8 weeks were randomly divided into two groups: (a) high cholesterol group (n=6), which is maintained on high-cholesterol diet for 6 weeks, and (b) niacin group (n=6), which receives the same cholesterol diet plus niacin (200 mg/kg/day) for 6 weeks. The control group (n=6) was fed with normal diet for 14 weeks. Subcutaneous adipose was collected for RNA analysis. The direct effect of niacin on leptin release was assayed in hypercholesterolemic rabbit adipocytes. Leptin levels in serum and adipocyte culture supernatant were measured via enzyme-linked immunosorbent assay. RT-PCR was used to evaluate leptin, PPARgamma, and CD36 mRNA expression in adipose and adipocytes. RESULTS: Compared with the control group, rabbits fed with high-cholesterol diets showed higher levels of serum total cholesterol, low-density lipoprotein cholesterol, and leptin, all of which were significantly reduced by niacin treatment. After 6 weeks of treatment with niacin, the leptin level was significantly decreased by 21.8% (6.87+/-1.58 vs. 8.79+/-1.45, P<.05) and leptin mRNA expression of adipose was significantly lower in rabbits treated with niacin than in those fed with high-cholesterol diet continuously (0.58+/-0.11 vs. 0.73+/-0.15, P<.05). Niacin dose-dependently inhibited leptin secretion and increased CD36 and PPARgamma expression in cultured adipocytes. The reduction of leptin mRNA expression of hypercholesterolemic rabbits by niacin was negatively correlated with the up regulation of PPARgamma and CD36 mRNA expression by niacin (r=-.69 and r=-.63, respectively, P<.01). CONCLUSION: Niacin can reduce serum level and adipose mRNA expression of leptin and up-regulate PPARgamma and CD36 mRNA expression in hypercholesterolemic rabbits. PMID- 18402820 TI - Effects of overexpansion on stents' recoil, symmetry/asymmetry, and neointimal hyperplasia in aortas of hypercholesterolemic rabbits. AB - BACKGROUND: It is not known whether overexpansion modifies stent recoil, symmetric distribution of struts, and neointimal hyperplasia. OBJECTIVES: The objectives were (a) to evaluate whether stent overexpansion modifies the geometric configuration of the stent in the arterial wall, (b) to determine the relationship between overexpansion and stent recoil, and (c) to evaluate the relationship between the distribution of struts and neointimal hyperplasia. METHODS: Twenty tubular stainless steel 316L stents (3.0 and 3.5 mm in diameter) were implanted at 20 and 10 atm, respectively, in the abdominal aorta of New Zealand rabbits fed a hypercholesterolemic diet (1% cholesterol). Sham operations were also performed in seven animals. Eight weeks after implantation or sham operation, an intravascular ultrasound (IVUS) study was performed to measure stent recoil and aid in stent classification (symmetric or asymmetric) according to strut distribution. The degree of injury and neointimal hyperplasia were also evaluated in hematoxylin-eosin stained sections. RESULTS: The symmetry/asymmetry of stents assessed by IVUS, as well as the neointimal hyperplasia, was similar in both groups. Stent recoil was significantly greater in the 3.0-mm stent (overexpanded) group (0.28+/-0.02 mm), as compared with stent recoil in the 3.5 mm stent group (0.10+/-0.01 mm, P<.05). The neointimal hyperplasia in histological slices, independent of the implant technique, was predominantly in zones with higher strut concentration as compared with zones with fewer struts. CONCLUSIONS: Stent overexpansion enhanced stent recoil and did not modify symmetric and asymmetric strut distribution. Neointimal hyperplasia was not modified by the implant technique. Interestingly, significant hyperplasia was observed in locations with greater strut concentration, independent of overexpansion. PMID- 18402821 TI - Isolated pericardial and intracardiac hydatidosis: presentation as congestive cardiac failure and fatal pulmonary embolism. AB - Localization of hydatid cysts in the heart is a rare phenomenon, with an incidence of 0.5-2%. In almost half these cases, the heart is the sole organ to be involved. We report a case of massive pericardial hydatidosis in a female patient who presented with features of congestive cardiac failure. Cysts in the endocardium of right side of the heart resulted in fatal pulmonary embolism. PMID- 18402822 TI - Detection of infectious agents by polymerase chain reaction in human aortic wall. AB - INTRODUCTION: Several studies have been suggested that infectious agents may induce or progress the process of atherosclerosis in humans. In the present study, the samples of visually healthy human aortic wall were examined for the presence of Chlamydia pneumoniae, Mycoplasma pneumoniae, Helicobacter pylori, herpes simplex virus (HSV), and cytomegalovirus (CMV). METHODS: Bacterial DNA of C. pneumoniae, M. pneumoniae, and H. pylori and viral DNA of HSV and CMV were analyzed by polymerase chain reaction. The specimens were obtained from 40 patients with atherosclerotic three-vessel stable coronary artery disease referred to surgical revascularization (coronary group) and 20 controls referred to aortic valve replacement (valve group). RESULTS: C. pneumoniae was detected in 11 of 40 samples of aorta in coronary group (27.5%) and 5 of 20 in valve group (25%). M. pneumoniae was found in 6 of 40 (15%) and 5 of 20 (25%) samples, and CMV was found in 22 of 40 (55%) and 10 of 20 (50%) samples. The most frequent detected pathogens were H. pylori and HSV. H. pylori was found in 32 of 40 samples of aortic wall in coronary group (80%) and 17 of 20 samples in valve group (85%), whereas HSV was found in 27 of 40 (67.5%) and 17 of 20 (85%) aortic wall specimens. CONCLUSION: Results demonstrate that C. pneumoniae, M. pneumoniae, H. pylori, CMV, and HSV can be detected in macroscopically healthy aortic wall of coronary and valve patients in similar frequency, which do not support hypothesis concerning the role of microorganisms in atherosclerosis etiology. PMID- 18402823 TI - Redo mitral valve surgery: morphological features. AB - INTRODUCTION: Mitral valve (MV) repair is the surgery of choice in patients with mitral regurgitation. This study reviews the histological features of excised mitral leaflets and annuloplasty rings of patients who had previous MV repair or attempted repair. MATERIALS AND METHODS: This study examines the morphological findings of 54 excised MV leaflets and 39 annuloplasty rings, excised over 15 years at one institution. Patients were separated into groups based on the time interval between surgeries: attempted repair, <30 days, 30-365 days, and >1 year. RESULTS: The fifty-four patients had a mean age of 55.7+/-17.9 years at the time of surgery after failed repair or attempted repair, of the MV. Index surgery for all patients had been performed between 1973 and 2005. The interval between index surgery and readmission (n=49, 91%, excluding those with MV replacement following attempted repair) was 6.61+/-7.12 years. Myxomatous change and infective endocarditis (IE) were found in valves with failed attempted MV repair. Nonleaflet (ischemic, chordal rupture, and flow abnormalities) and acquired changes, degenerative, myxomatous, and IE were found in valves repaired in patients at <30 days postoperatively. Degenerative changes (myxomatous and RF) were the main underlying pathology found in valves repaired between 31 and 365 days previously. At over 1 year after original surgery, repairs were found to have congenital, degenerative (myxomatous, rheumatic valvular disease, and IE) and nonleaflet (ischemic and chordal rupture) changes. CONCLUSIONS: Although MV repair is the preferred treatment for mitral regurgitation, failure of repair may occur in patients at the time of surgery or over varying periods after surgery. The pathological findings following repair include IE, fibrosis, calcification, and nodular thickening at and around the operative site, thickening and stiffening of the synthetic chordae tendinae due to tissue overgrowth, rupture of chordae tendinae, and progression of acquired degenerative disease. PMID- 18402824 TI - Association of echocardiographic atrial size and atrial fibrosis in a sequential model of congestive heart failure and atrial fibrillation. AB - BACKGROUND: Cardioversion (CV) success of atrial fibrillation (AF) inversely correlates to the size of the left atrium (LA). Atrial fibrillation and its most important risk factor, congestive heart failure (CHF), both induce atrial structural enlargement and fibrosis. To investigate the effect of AF and CHF on atrial dilatation and fibrosis, and to estimate whether echocardiographically determined atrial size may be used as a marker for atrial fibrosis. METHODS: In six dogs, pacemakers were implanted followed by HIS bundle ablation. After 4 weeks of rapid ventricular stimulation (185 bpm) for CHF induction, additional rapid atrial stimulation (500 bpm) was maintained for 7 weeks to induce AF. Serial determinations of echocardiographic atrial size were performed. Seven dogs with sinus rhythm served as histological controls. Postmortem tissue was obtained to determine the degree and composition of atrial fibrosis. RESULTS: While the ejection fraction of the AF/CHF dogs decreased significantly from 57+/-5% to 19+/ 7% (P<.01), an increased degree of atrial fibrosis was found (right atrium [RA], 4.9+/-2.0% to 19.9+/-5.4%; LA, 4.4+/-1.6% to 22.2+/-3.2%; P<.01), accompanied by a significant increase of atrial volumes (LA: 21+/-4 to 44+/-4 mm3; P<.01; RA: 10+/-3 to 18+/-6 mm3; P<.05) and LA diameters (34+/-4 to 43+/-2 mm, P<.05). Atrial fibrosis and size significantly correlated. CONCLUSIONS: Atrial fibrillation/CHF leads to a significant atrial fibrosis and dilation. The increased echocardiographic size correlates to the degree of atrial fibrosis and may be used as clinical marker for atrial fibrosis. The fibrosis accompanying atrial dilatation may also explain why LA size, as determined by echocardiography, is a strong predictor of CV success. PMID- 18402825 TI - Ascending aortic aneurysms in unicommissural aortic valve disease. AB - BACKGROUND: Aneurysms of the ascending aorta occur as result of intrinsic changes in the aortic wall and have been well documented in patients with bicuspid aortic valve (BAV). In few reported clinical studies, documenting aneurysmal dilatation in unicommissural aortic valves (UAV); there have been no comments on the aortic wall pathology. This study presents the pathological findings of the ascending aorta in patients with UAV. MATERIALS AND METHODS: The clinical data from 39 patients with concomitant excision of the UAV and aneurysmal aortic tissue were reviewed. In all cases, the gross features of the valve and aortic segments were noted and submitted for histology. The sections of the aorta were semi quantitatively graded for the extent of medionecrosis, cystic medial change, fibrosis, and elastic tissue changes (fragmentation/ loss) in the media. The medial alterations were correlated with patient age, gender, and valvular dysfunction, and compared to aneurysmal disease in BAV and three-cuspid aortic valves (TAV) excised over a 3-year period. RESULTS: Among 39 patients studied, a majority were males (92.3%), with a mean age at surgery of 39.92 years. Only three patients (7.69%) were above the age of 50 years. Eighteen patients (46.1%) had aortic stenosis with regurgitation. Ascending aorta diameters ranged from 4 to 5.5 cm. The overall pattern of medial changes was nearly the same in all cases of UAV, irrespective of age and nature of valvular dysfunction. Most cases showed mild histological changes, with medionecrosis and fibrosis being the more common and consistent features. However, varying grades of change affected different portions of the media and/or the aortic wall in the same patient. The changes in UAV aortae were comparable to the changes seen in the TAV and BAV, but these differed with the age of onset. CONCLUSIONS: This study demonstrates the presence of medial changes in the ascending aortic tissue in all patients of UAV with aneurysms. These changes, while mild to moderate in degree, likely have a similar pathogenetic mechanism as those seen in BAV disease. The significant difference in age, at the time of surgery, suggests a more rapid progression of the aortic changes. PMID- 18402826 TI - An unusual right heart mass. AB - A mobile right atrial mass, prolapsing into the right ventricle and extending to the inferior vena cava, was unexpectedly discovered during a routine echocardiographic examination in a 53-year- old woman. A possible tumour was suspected, but no evidence of primary malignancy was detected in lungs, liver, kidneys and adrenal glands. The patient underwent surgery, and only histological examination of the mass provided a final diagnosis of thrombosis. PMID- 18402827 TI - Pericardial patch repair of the left atrioventricular valve in atrioventricular septal defect: long-term changes in the patch. AB - A 31-year-old woman with partial atrioventricular septal defect underwent left atrioventricular valve (LAVV) replacement. Her initial repair was at 8 years of age. At 23 years of age, she underwent reoperation due to a combination of severe left ventricular outlet obstruction and moderate LAVV regurgitation. At that reoperation, she had a Dacron patch enlargement of the infundibular septum and repair of her LAVV with a xenograft (bovine) pericardial patch sutured into the superior bridging leaflet. LAVV replacement was required 8 years later because of valve insufficiency. There was a perforation in the patch with fibrosis, thickening due to pannus, and calcification of the pericardial tissue and the leaflet tissue, leading to stiffening of the tissue. PMID- 18402828 TI - Endometrial adenocarcinoma metastatic to the right ventricle: a case report and review of the literature. AB - BACKGROUND: Endometrial adenocarcinoma of any histologic type rarely metastasizes to the heart. Only three such metastases to the myocardium and three to the pericardium have been reported antemortem in the literature. The rarity of this metastasis is likely explained by the relative avascularity of the endocardium and velocity of blood. METHODS AND RESULTS: A 62-year-old female was admitted for increasing shortness of breath over the previous month. Her past medical history was significant for a resected Stage IC endometrial adenocarcinoma endometrioid type 15 months prior. The tumor was found to be a poorly differentiated (Grade 3) endometrial adenocarcinoma invading over half the myometrium. The periaortic and pelvic lymph nodes as well as the cervix and pelvic organs were free of tumor. The patient was later treated with adjuvant external beam radiotherapy. An echocardiogram demonstrated a large right ventricular mass. Subsequent endocardial biopsy showed a poorly differentiated, Grade 3, endometrial adenocarcinoma of endometrioid histologic type. The tumor was deemed inoperable given its size and vast involvement of the myocardium so palliative care was provided and the patient expired 17 days later. CONCLUSION: Management of metastatic adenocarcinoma to the heart is not well established due to the rarity of this lesion. Previously reported cases vary in the therapeutic approach as well as the outcome. To this date the best outcome has been a survival of 6 years after treatment of the metastasis with radiotherapy and concurrent cisplatin and pegylated liposomal doxorubicin. PMID- 18402830 TI - Right ventricular endomyocardial fibrosis. AB - Endomyocardial fibrosis (EMF) is a neglected tropical cardiomyopathy of unknown etiology and pathogenesis that is common in certain tropical areas of Africa, Asia, and South America. It affects predominantly children and young adults. Endocardial fibrosis is the hallmark of this restrictive cardiomyopathy leading to restriction to diastolic filling of the ventricles with severe atrial dilatation. Endomyocardial fibrosis carries a poor prognosis, due to the late presentation of patients. The salient features of this condition are present in the case presented here of a 14-year-old boy who died from progressive heart failure due to right ventricular EMF. These pathological findings of advanced disease highlight the need for early diagnosis and better understanding of the pathogenesis in order to improve prognosis of this debilitating and fatal disease. PMID- 18402829 TI - Cardiotin localization in mitochondria of cardiomyocytes in vivo and in vitro and its down-regulation during dedifferentiation. AB - BACKGROUND: Cardiotin expression is observed in adult cardiac tissue. In the present study, we provide evidence for the specific localization of cardiotin in cardiac mitochondria and for its down-regulation during adaptive remodeling (dedifferentiation) of cardiomyocytes. METHODS: Immunocytochemistry was used to study cardiotin localization in adult rabbit papillary muscle, in late-stage embryonic rabbit left ventricular tissue, and in left ventricle samples of rabbits suffering from pressure and volume overload. Western blot analysis of cardiotin was performed in purified pig heart mitochondrial fractions. Cardiotin expression was monitored in vitro in isolated adult rat and rabbit left ventricular cardiomyocytes. RESULTS: Western blot analysis revealed the presence of cardiotin in the mitochondrial fractions of pig heart. Immunoelectron microscopy confirmed the presence of cardiotin in cardiac mitochondria of normal adult rabbits both in vivo and in vitro. Quantification of the localization of immunogold particles suggests an association of cardiotin with the mitochondrial inner membrane. Cardiotin expression is initiated in late-stage embryonic rabbit heart, whereas in adult ventricular tissue cardiotin clearly stained longitudinal arrays of mitochondria. Pressure- and volume-overloaded myocardium showed a reduction in cardiotin expression in dispersed local myocardial areas. Cell cultures of adult cardiomyocytes showed a gradual loss in cardiotin expression in parallel with a sarcomeric remodeling. CONCLUSIONS: Our results demonstrate the specific localization of cardiotin in adult cardiomyocyte mitochondria and propose its use as an early marker for cardiomyocyte adaptive remodeling and dedifferentiation. PMID- 18402831 TI - Primary bi-atrial Burkitt lymphoma with severe inflow impairment in an immunocompetent patient. AB - We report herein a case of sporadic primary cardiac bi-atrial Burkitt lymphoma (BL) occurred in a 67-year-old white immunocompetent patient and presenting with signs and symptoms of severe bilateral atrioventricular inflow impairment. Extranodal BL involving the heart is rare and seldom recognized clinically. Delayed discovery contributes to significant mortality. In the case presented extended surgical excision and intensive combination chemotherapy regiments resulted in complete remission at 1 year. PMID- 18402832 TI - Coronary atherosclerotic lesions in human immunodeficiency virus-infected patients: a histopathologic study. AB - BACKGROUND: Studies suggest human immunodeficiency virus-positive (HIV+) patients have an increased risk of coronary artery disease (CAD), yet little is known about the histopathology, severity, or distribution of lesions. METHODS: The coronary arteries of 66 deceased AIDS patients and 19 HIV controls (age <55) were dissected and graded for percent luminal stenosis by intimal lesions, percent of intima involved with lipid, and extent of intimal calcification on a scale of 0 to 3. Medical histories, antiretroviral therapies, and CAD risk factors were reviewed. RESULTS: HIV+ patients were older than controls (P=.06), and more were male (P=.02). Thirty-five percent of HIV+ patients had stenosis >or=75% of at least one artery. Compared to controls, HIV+ patients had three times greater odds of stenosis >or=75%, controlling for age and sex (one-sided P=.03). Older age and male sex were also risk factors (one-sided P<.001). HIV seropositivity was associated with increased plaque lipid content (one-sided P=.02) and calcification (one-sided P=.08). Duration of HIV infection, antiretroviral therapy, and immune status did not predict severe disease in multivariate analyses. Previously unreported patterns of dystrophic calcification were observed in HIV+ patients and older controls. CONCLUSIONS: Young to middle-aged patients dying from advanced AIDS have atherosclerotic CAD that may result in luminal narrowing, heavy calcification, and high plaque lipid content. The pattern of disease, location of lesions, and plaque composition are typical of atherosclerosis in HIV-negative patients. No relationship between antiretroviral therapies and atherosclerosis was seen in this small study of heavily treated patients. PMID- 18402833 TI - Increased matrix metalloproteinase-8 and -9 activity in patients with infarct rupture after myocardial infarction. AB - BACKGROUND: Infarct rupture is a usually fatal complication of myocardial infarction (MI), for which no molecular mechanism has been described in humans. Experimental evidence in mouse models suggests that the degradation of the extracellular matrix by matrix metalloproteinases (MMPs) plays an important role in infarct rupture. The present study was designed to study the role of MMP-2, MMP-8, and MMP-9 in human infarct rupture. METHODS: Heart samples were obtained from patients who died from infarct rupture and control MI patients. The MMP activity was determined by zymography and quantitative immunocapture activity assay. TIMP-1 levels were measured and immunohistochemistry for MMP-2 and MMP-9 was performed. RESULTS: The amounts of both total and active MMP-8 and MMP-9 were significantly higher in ruptured infarct tissue than in control MI tissue, but no differences in MMP-2 activity were observed. Furthermore, the number of inflammatory cells was significantly higher in the ruptured infarcts than in control infarcts. CONCLUSIONS: These data suggest that increased MMP-8 and MMP-9 activity in the infarct area, caused by a more prominent infiltration of inflammatory cells, contribute to infarct rupture in humans. PMID- 18402834 TI - Cardiac oxidative stress and remodeling following infarction: role of NADPH oxidase. AB - BACKGROUND: There is growing recognition that oxidative stress plays a role in the pathogeneses of myocardial repair/remodeling following myocardial infarction (MI). Nicotinamide adenine denucleotide phosphate (NADPH) oxidase is a major source for cardiac reactive oxygen species production. Herein, we studied the importance of NADPH oxidase in development of cardiac oxidative stress and its induced molecular and cellular changes related to myocardial repair/remodeling. METHODS: MI was created by coronary artery ligation in C57/BL (wild type) and NADPH oxidase (gp91(phox)) knockout mice. Cardiac oxidative stress, inflammatory/fibrogenic responses, apoptosis, and hypertrophy were detected by in situ hybridization, immunohistochemistry, terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL), picrosirius red staining, and image analysis, respectively, at different stages post MI. RESULTS: In wild-type mice with MI, and compared to sham-operated animals, we observed significantly increased gp91(phox) and 3-nitrotyrosine, a marker of oxidative stress, in the infarcted myocardium; accumulated macrophages and myofibroblasts at the infarct site; abundant apoptotic myocytes primarily at border zones on Day 3; and numerous apoptotic inflammatory/myofibroblasts in the later stages. In addition, we detected significantly increased transforming growth factor beta1, tissue inhibitor of metalloprotease 2, and type 1 collagen gene expression; continuously increasing collagen volume in the infarcted myocardium; and hypertrophy in noninfarcted myocardium. Compared to wild-type mice with MI, we did not observe significant difference in infarct size/thickness, cardiac hypertrophy, myocyte apoptosis, inflammatory/fibrogenic responses, as well as cardiac oxidative stress in gp91(phox) knockout mice. CONCLUSION: Our findings indicate that during NADPH oxidase deficiency, superoxide production can be compensated by other sources, which leads to cardiac oxidative stress and its related molecular/cellular events in the infarcted heart. PMID- 18402835 TI - Identification of tissue factor in experimental aortic valve sclerosis. AB - BACKGROUND: Aortic valve sclerosis (AVS) shares epidemiological and histological similarities with atherosclerosis. Tissue factor (TF), the main initiator of blood coagulation, is present in atherosclerotic plaques and contributes to their thrombogenicity. We aimed to analyze valvular TF expression in addition to other components of atherosclerosis in two models of AVS. METHODS: Forty-five rabbits were randomly assigned to receive either normal chow (Ctrl, n=15), or 1% cholesterol-enriched chow alone (Hyperchol, n=15) or associated with vitamin D(2) (VitD, n=15), for 12 weeks. Aortic valve (AV) performance, leaflet structure, cellular and lipid infiltration, and TF expression were assessed using Doppler, histology, and immunohistochemistry, respectively, and TF activity was evaluated in AV leaflets. RESULTS: Hyperchol and VitD animals developed abnormal leaflet thickening, with a significant alteration of AV performance in VitD animals. Leaflet thickening was related to the development of fatty plaque neolesions on the aortic side of the leaflets, displaying extracellular matrix disorganization, lipid and cellular infiltration, and calcification in VitD animals. TF was found on the leaflet aortic side in Ctrl animals and was identified in AVS lesions in both Hyperchol and VitD animals. TF immunostaining area and valvular activity increased significantly across the three groups. CONCLUSIONS: Experimental AVS lesions that are present on the aortic side of leaflets display numerous characteristics of vascular atherosclerosis, including TF expression. Identification of TF associated with other components of the atherosclerotic process in AVS lesions strengthens the link between atherosclerosis and AVS. PMID- 18402836 TI - Hypothyroidism-induced myocardial damage and heart failure: an overlooked entity. AB - BACKGROUND: Hypothyroid state may induce cardiac muscle impairment such as diastolic dysfunction and abnormal relaxation time. Advanced heart failure in hypothyroid patients has been described only in severe symptomatic cases, mostly during myxedematous coma. METHODS AND RESULTS: We describe an unusual case of asymptomatic patient with hypothyroidism who presented with severely reduced cardiac function with elevated cardiac enzymes reflecting significant myocardial injury. Comprehensive evaluation for heart failure was suggestive only for long standing untreated hypothyroidism. Endomyocadial biopsy demonstrated unique histological findings of mucopolysaccharide accumulation attributed to hypothyroid state. CONCLUSIONS: Asymptomatic hypothyroidism may cause severe reduction in cardiac function accompanied with elevated cardiac enzymes. To our knowledge, this is the first description of human myocardial biopsy revealing mucopolysaccharide accumulation attributed to hypothyroid state. PMID- 18402837 TI - Sudden death in tuberculous myocarditis. AB - Tuberculous myocarditis is one of the rare causes of sudden death. We report a case of 65-year-old female who came with diabetic foot and died suddenly after 2 days of hospital stay. On autopsy, she was found to have tuberculous myocarditis with granulomatous inflammation only in the liver and without pulmonary or mediastinal lymph node involvement. These findings are very unusual in tuberculous myocarditis. PMID- 18402838 TI - Verrucoid lesions of mitral valve in a dog with features of inflammatory myofibroblastic tumor. AB - We report a mitral valve lesion detected at autopsy in a 9-year-old male German Shepherd dog suffering from mild mitral regurgitation. Gross examination of the heart showed exophitic, noncontiguous lesions involving the atrial aspect of both mitral leaflets. Microscopic evaluation of the mitral lesions disclosed a diffuse proliferation of myofibroblasts with little atypia, arranged loosely and rather randomly, within a myxoid stroma and associated with inflammatory cells identified as CD138+ plasma cells, CD68+ macrophages, and eosinophils. The myofibroblastic proliferation we describe is quite similar to previously described inflammatory myofibroblastic tumor (IMT), a very rare lesion, exceptionally found in the human heart. PMID- 18402839 TI - Role of carbonic anhydrase II in ectopic calcification. AB - INTRODUCTION: Osteopontin (OPN) is a potent inhibitor of ectopic calcification. Previous studies suggested that, in addition to blocking apatite crystal growth, OPN promoted regression of ectopic calcification by inducing the expression of acid-generating carbonic anhydrase II (CAR2) in monocyte-derived cells. METHODS: To test this hypothesis, OPN and CAR2 expression and calcification of subcutaneously implanted glutaraldehyde-fixed bovine pericardium (GFBP) were studied in CAR2 mutant mice. RESULTS: Consistent with previous studies in Black Swiss mice, GFBP calcified to a greater extent in OPN-deficient mice compared to wild types on the C57Bl/6 background. GFBP implanted in CAR2-deficient mice (CAR2(-/-)) were significantly more calcified than those implanted into wild-type mice (CAR2(+/+)) [37+/-5 vs. 20+/-6.5 microg Ca/mg tissue, respectively, at 30 days (P<.001), and 42+/-5 versus 20+/-4 microg Ca/mg tissue at 60 days, respectively (P<.001)]. On the other hand, OPN levels within and surrounding the implants were similar in CAR2(+/+) and CAR2(-/-) mice, suggesting that OPN expression in the absence of CAR2 was not sufficient to mitigate ectopic calcification. CONCLUSIONS: These results indicate that CAR2 expression is an important regulator of ectopic calcification, potentially by facilitating OPN mediated mineral regression. PMID- 18402840 TI - Local mechanical and structural properties of healthy and diseased human ascending aorta tissue. AB - OBJECTIVE: This study investigates the mechanics and histology of healthy and dilated human ascending aortas (AA). The regional variation in mechanical response and tissue structure were compared. METHODS: Rings of human AA from healthy (n=5), dilated tricuspid aortic valve (TAV, n=5), and dilated bicuspid aortic valve (BAV, n=6) patients were mechanically tested. Each aortic ring was sectioned into quadrants-anterior, posterior, medial (inner curvature) and lateral (outer curvature). Low- and high-stress elastic moduli were calculated from the equibiaxial stress strain curve to determine the local mechanical properties. Histological analysis was used to quantify the percent composition of elastin, collagen, and smooth muscle cells. RESULTS: BAV tissue was thinnest and contained the largest percent composition of collagen. Both TAV and BAV tissue had significantly less elastin than healthy tissue. At low strain in the circumferential direction, TAV tissue was on average the least stiff. The elastic modulus was dependent on quadrant and tissue type but not direction (isotropic). Generally, the lateral quadrant tissue was the stiffest and the medial quadrant the least stiff. There were no apparent local variations in the tissue histology. CONCLUSIONS: Local variations in tissue thickness and mechanical properties were evident in all samples analyzed and may be linked to the type of aortic valve present. PMID- 18402841 TI - Primary cardiac lymphoma: molecular cytogenetic characterization of a rare entity. AB - BACKGROUND: The majority of cardiac atrial neoplasms represent benign myxomas. Rarely, malignant cardiac neoplasms are encountered and can include primary cardiac neoplasms, as well as secondary tumors involving the heart. As many cardiac neoplasms lack pathognomonic clinical features, histopathologic diagnosis is crucial for classification and appropriate treatment of these neoplasms. Molecular investigation is critical to begin to catalogue genomic changes that correlate with these malignancies. METHODS: A 60-year-old man presented with superior vena cava syndrome, and computed tomographic scan revealed an infiltrative mass of the right atrium that nearly filled the atrial chamber and partially occluded superior vena cava flow. Urgent surgical resection revealed a soft mass with the appearance of "fish flesh." Histologic, immunochistochemical, cytogenetic, and detailed molecular investigations were carried out. RESULTS: Histologic examination revealed complete replacement of the atrial wall by diffuse sheets of pleomorphic lymphoid cells with occasional smaller plasmacytoid cells. The predominant lymphoid population was immunoreactive for CD45, CD20, CD79a, BCL-2, BCL-6, Ki-67, CD10, p53, and light chain restricted for IgM lambda. A diagnosis of primary cardiac diffuse large B-cell lymphoma with plasmacytoid differentiation was established and was supported by cytogenetic studies demonstrating the presence of a t(14;18)(q32;q21) translocation in addition to other chromosomal abnormalities. Fluorescence in situ hybridization revealed no evidence of a C-MYC translocation. CONCLUSION: In this single case, comparative genomic hybridization analysis using both bacterial artificial chromosome and oligonucleotide arrays correlated well with cytogenetic findings and allows for the cataloguing of more subtle genomic events. PMID- 18402842 TI - Cardiomyocyte death and renewal in the normal and diseased heart. AB - During post-natal maturation of the mammalian heart, proliferation of cardiomyocytes essentially ceases as cardiomyocytes withdraw from the cell cycle and develop blocks at the G0/G1 and G2/M transition phases of the cell cycle. As a result, the response of the myocardium to acute stress is limited to various forms of cardiomyocyte injury, which can be modified by preconditioning and reperfusion, whereas the response to chronic stress is dominated by cardiomyocyte hypertrophy and myocardial remodeling. Acute myocardial ischemia leads to injury and death of cardiomyocytes and nonmyocytic stromal cells by oncosis and apoptosis, and possibly by a hybrid form of cell death involving both pathways in the same ischemic cardiomyocytes. There is increasing evidence for a slow, ongoing turnover of cardiomyocytes in the normal heart involving death of cardiomyocytes and generation of new cardiomyocytes. This process appears to be accelerated and quantitatively increased as part of myocardial remodeling. Cardiomyocyte loss involves apoptosis, autophagy, and oncosis, which can occur simultaneously and involve different individual cardiomyocytes in the same heart undergoing remodeling. Mitotic figures in myocytic cells probably represent maturing progeny of stem cells in most cases. Mitosis of mature cardiomyocytes that have reentered the cell cycle appears to be a rare event. Thus, cardiomyocyte renewal likely is mediated primarily by endogenous cardiac stem cells and possibly by blood-born stem cells, but this biological phenomenon is limited in capacity. As a consequence, persistent stress leads to ongoing remodeling in which cardiomyocyte death exceeds cardiomyocyte renewal, resulting in progressive heart failure. Intense investigation currently is focused on cell based therapies aimed at retarding cardiomyocyte death and promoting myocardial repair and possibly regeneration. Alteration of pathological remodeling holds promise for prevention and treatment of heart failure, which is currently a major cause of morbidity and mortality and a major public health problem. However, a deeper understanding of the fundamental biological processes is needed in order to make lasting advances in clinical therapeutics in the field. PMID- 18402843 TI - Primary cardiac osteosarcoma with recurrent episodes and unusual patterns of metastatic spread. AB - Primary osteosarcoma of the heart is an extremely rare entity. In this report, we describe a case of primary osteosarcoma of the heart that recurred and metastasized. The patient is a 50-year-old woman who presented with an abrupt onset of dyspnea, dizziness, and palpitations. Echocardiography results showed a left atrial tumor that was resected and had histological features of high-grade osteosarcoma. The patient was treated with adjuvant chemotherapy. The tumor recurred in the same location 4 years later and was resected. Three years later, the patient presented with a 4-cm polypoid mass in the stomach that was consistent with metastatic osteosarcoma. One year later, the patient presented with recurrent high-grade sarcoma in the soft tissue of the left chest wall. We herein present the spectrum of histological findings and molecular genotyping data. PMID- 18402844 TI - Whole-body hyperthermia attenuates experimental autoimmune myocarditis in the rat. AB - BACKGROUND: Heat stress prior to induction of various forms of cardiac injury has been shown to result in preconditioning and attenuation of subsequent damage. We evaluated the effects of whole-body hyperthermia (WBH) on experimental autoimmune myocarditis (EAM) induced either by injection of myosin or by adoptive transfer of lymphocytes. METHODS AND RESULTS: Lewis rats were pretreated with WBH either 24 h prior to EAM induction (Group A1) or 14 days following EAM induction (Group A2). The control group included myocarditic rats that were not exposed to WBH (Group A3). Rats from Group A1 exhibited significant protection from myocarditis as compared to rats from Group A3, evidenced by reduced myocarditis scores (1.60+/-0.96 vs. 2.88+/-0.35, P=.016). Rats from Group A2 also exhibited protection from myocarditis although not significantly. In a second experiment, we used adoptive transfer of myosin-reactive lymphocytes to study the mechanism of WBH effect on myocarditis. There was evidence of microscopic myocarditis in four out of five rats that were injected with active lymphocytes (the Control Group B3). Myocarditis was not observed in rats adoptively transferred with preheated rat lymphocyes (Group B1) nor in preheated rats, which underwent administration of nonheated rat lymphocyes (Group B2). CONCLUSIONS: Whole-body hyperthermia attenuates experimental myocarditis in the rat. The beneficial effect of whole-body hyperthermia may be related to immunomodulatory effect and direct cardiomyocyte protection. PMID- 18402845 TI - Self-administration of injectable contraceptives. AB - Although depot-medroxyprogesterone represents a highly effective contraceptive, its use is associated with poor continuation rates. Although the major reason for discontinuation is menstrual irregularity, the time, expense and inconvenience of clinic visits also pose a barrier to use. Self-administration of the 104-mg subcutaneous formulation might make clinic visits unnecessary. Many medications can be safely self-administered by subcutaneous injection, and patient satisfaction is high. Appropriate patient selection, adequate training, use of prefilled injection devices and counseling regarding bleeding patterns are likely to maximize success rates with self-administration. By improving the convenience of this contraceptive method, self-injection might improve both compliance and continuation rates. The potential for self-administration of this contraceptive deserves formal study. PMID- 18402846 TI - Copper-containing, framed intrauterine devices for contraception: a systematic review of randomized controlled trials. AB - BACKGROUND: Intrauterine devices (IUDs) are safe and effective methods of long term reversible contraception. The design and copper content as well as placement of the copper on IUDs could affect their effectiveness and side effect profile. We compared different copper IUDs for their effectiveness and side effects. STUDY DESIGN: We searched multiple electronic databases with appropriate keywords and names of the IUDs known to be on the market. We searched the reference lists of papers identified and contacted authors when possible. There was no language restriction. Randomized controlled trials comparing different IUDs that reported on clinical outcomes were considered for inclusion. Two reviewers independently extracted data on outcomes and trial characteristics. We combined the trial results in meta-analyses and expressed results as rate difference (RD) using a fixed-effects model with 95% confidence interval (CI). In the presence of significant heterogeneity, a random-effects model was applied. RESULTS: We included 35 trials, resulting in 18 comparisons of 10 different IUDs in approximately 48,000 women. TCu380A was more effective in preventing pregnancy than MLCu375 (RD 1.70%, 95% CI 0.07-2.95% after 4 years of use). TCu380A was also more effective than MLCu250, TCu220 and TCu200. There tended to be fewer pregnancies with TCu380S compared to TCu380A after the first year of use, a difference which was statistically significant in the fourth year (RD -1.62%, 95% CI -3.00% to -0.24%). This occurred despite more expulsions with TCu380S (RD 3.50%, 95% CI 0.36-6.63% at 4 years). MLCu375 was no more effective than TCu220 at 1 year of use, or MLCu250 and NovaT up to 3 years. Compared to TCu380A or TCu380S, none of the IUDs showed any benefits in terms of bleeding or pain or any of the other reasons for early discontinuation. None of the trials that reported events at insertion found one IUD easier to insert than another or caused less pain at insertion. There is no evidence that uterine perforation rates vary by type of device. There are minimal randomized data on IUD use in nulliparous women. CONCLUSIONS: TCu380A and TCu380S appear to be more effective than other IUDs. No IUD showed consistently lower removal rates for bleeding and pain in comparison to other IUDs. There is no evidence that any particular framed copper device is better suited to women who have not had children. PMID- 18402847 TI - Levonorgestrel administration in emergency contraception: bleeding pattern and pituitary-ovarian function. AB - BACKGROUND: This study was conducted to evaluate the effects of levonorgestrel administration for emergency contraception (EC) on bleeding pattern and pituitary ovarian function. STUDY DESIGN: In 69 women with a reported stable menstrual cycle length of 24-34 days, we investigated bleeding patterns following EC administration in the follicular (n=26), periovulatory (n=14) and luteal (n=29) phase. In a subgroup of 8 women, hormonal evaluation and ultrasonography were performed. RESULTS: EC taken in the follicular, but not in the periovulatory or luteal phase, significantly shortened cycle length by 10.9+/-1 days. The subsequent cycle was not affected. EC taken in the late preovulatory phase, prior to the gonadotrophin surge, suppressed ovulation (n=7), while ovulation was not blocked when EC was given during an ongoing luteinizing hormone (LH) pulse (n=1). CONCLUSIONS: Our data indicate that EC given before the onset of the luteinizing hormone (LH) surge inhibits ovulation and hastens the end of the current menstrual cycle. Subsequently, the length of the following menstrual cycle returned as prior to treatment. By contrast, levonorgestrel administered after the expected ovulation has no effect on menstrual cycle length. PMID- 18402848 TI - Effect of an emergency contraceptive pill intervention on pregnancy risk behavior. AB - OBJECTIVE: Previous trials of emergency contraceptive pills (ECPs) found that in menstrual cycles ending in pregnancy, ECP use was more common among women exposed to interventions that enhanced access to the medication than among women with standard access. We examined data from one such trial to explore whether this finding has implications regarding the effect of the intervention on pregnancy risk behavior. METHODS: In our recent randomized trial, the intervention group received unlimited free ECPs in advance of need, whereas control participants obtained the medication when needed at usual cost. Participants were followed up for 1 year. In this secondary analysis, we examined ECP and contraceptive use in the cycles ending in pregnancy in that trial. RESULTS: Pregnancies in the intervention group appeared to have been more likely than those in the control group to be classified as "probably" or "possibly" ECP failures (12/74 in the intervention group vs. 1/74 in the control group; p=.012) and more likely to have occurred in the context of use of less efficacious contraceptives. CONCLUSION: Unrestricted access to ECPs in this trial may have increased the frequency of coital acts with the potential to lead to pregnancy. PMID- 18402849 TI - Efficacy, safety and sustainability of treatment continuation and results of an oral contraceptive containing 30 mcg ethinyl estradiol and 2 mg chlormadinone acetate, in long-term usage (up to 45 cycles)--an open-label, prospective, noncontrolled, office-based Phase III study. AB - BACKGROUND: This open-label, noncontrolled study assessed the long-term efficacy and tolerability of the monophasic combined low-dose oral contraceptive (OC) ethinyl estradiol (EE) 30 mcg+chlormadinone acetate (CMA) 2 mg (Belara). STUDY DESIGN: In total, 781 women who had already taken EE 30 mcg+CMA 2 mg for 24 cycles in a previous Phase III study were assessed for up to 45 cycles. RESULTS: Over 23,033 cycles, the Pearl Index was 0.16 (95% confidence interval, 0.04 0.42). Approximately 86% of women had regular withdrawal bleeding in each cycle, while incidence of intracyclic bleedings (1.6-6.4%) and proportion of women with amenorrhea (4%) were low. The incidence of acne decreased from 13.8% to 5.7%, while rates of hirsutism, alopecia and seborrhea remained low (< or =4%) throughout this study. The most frequent adverse events were consistent with OC treatment, and no unexpected events occurred. No changes in mean blood pressure and pulse rate were observed during the study, and there were no clinically relevant changes in liver or hematological parameters, hemostasis or carbohydrate metabolism. The incidence of pathological findings in gynecological examination was low and decreased over time. CONCLUSION: EE 30 mcg+CMA 2 mg was an effective and well-tolerated OC, with beneficial effects on cycle stability, intracyclic bleeding, amenorrhea and signs of androgenization that were maintained during long-term treatment for up to 5 years. There was no evidence of an increased risk of thromboembolic events, atherogenic disease or cervical cancer, suggesting that 30 EE mcg+CMA 2 mg is highly suitable for long-term use. PMID- 18402850 TI - Prospective study on the cytological aspects of the nasal respiratory epithelium in premenopausal women taking 30 mcg ethinylestradiol and 3 mg drospirenone oral contraceptive. AB - BACKGROUND: This study was conducted to determine the effects of the monophasic oral contraceptive (OC) containing 30 mcg ethinylestradiol (EE) and 3 mg drospirenone (DRSP) on the nasal respiratory epithelium in premenopausal women. STUDY DESIGN: Fifty premenopausal women participated in the prospective study. Baseline endovaginal ultrasound examination and blood test to measure serum progesterone to confirm an ovulatory cycle were performed. The cytologic changes on the nasal respiratory epithelium were evaluated with the maturation index, performed during the follicular, periovular and luteal phases of the menstrual cycle, and on the sixth cycle of pill intake. RESULTS: The maturation indexes of the nasal smears were higher during both the follicular and periovular phases than during the luteal phase of the menstrual cycle (p<.05). Women on the OC pill had similar changes in the nasal epithelium as those seen during the follicular phase (p NS), and less than the periovular phase (p<.05) and higher than the luteal phase (p<.05). The maturation index of the nasal respiratory epithelium seems to depend on the variation of the ovarian steroids during the menstrual cycle and on the iatrogenic effects of oral contraception. CONCLUSIONS: DRSP+EE seems to provoke cytological changes that are different from previously shown gestodene plus EE. PMID- 18402851 TI - Pregnancy during breastfeeding in rural Egypt. AB - BACKGROUND: Breastfeeding does not reliably protect against pregnancy except during the first 6 months postpartum and only then if accompanied by amenorrhea. Reluctance to use other methods of contraception during lactation may result in unplanned pregnancy. The aims of this study were to describe, among women in rural Egypt attending for antenatal care the prevalence of pregnancy during breastfeeding, contraceptive practice and unintended pregnancy. Finally, the study assessed women's impressions of the effect of conception during breastfeeding on breast milk and on the health of the breastfed infant. STUDY DESIGN: A descriptive study using an interviewer-administered structured questionnaire for 2617 parous women attending a hospital in Egypt for antenatal care. RESULTS: More than 95% of women breastfed the child before their current pregnancy; 25.3% conceived while breastfeeding. Conception occurred during the first 6 months postpartum in 4.4%, before resumption of menstruation in 15.1% and while exclusively or almost exclusively breastfeeding in 28.1%. Only 10 pregnancies (1.5%) occurred when all the prerequisites of the lactational amenorrhea method of contraception (LAM) were present. Twenty-nine percent of pregnancies conceived during breastfeeding were unintended, 10% of women had considered terminating their pregnancy while 4.4% of them reported trying to do so. CONCLUSIONS: Pregnancy during breastfeeding is common in Egypt and is often unintended. There is great potential for using LAM, but it must be properly taught, and women should be encouraged to start using effective contraception as soon as any of the prerequisites of LAM expires. PMID- 18402852 TI - Can intrauterine contraceptive devices be a Candida albicans reservoir? AB - BACKGROUND: The in vitro adherence of Candida albicans isolated from vaginal exudates of patients with vulvovaginal candidiasis (VVC) to intrauterine contraceptive devices (IUDs) and biofilm formation capacity were evaluated. STUDY DESIGN: This research was conducted with two vaginal C. albicans isolates. The adherence on IUD by both radiomarked adhesion assay and scanning electron microscopy, and the biofilm production capacity by spectrophotometric method were determined. RESULTS: The yeasts adhered strongly to different parts of the IUD (covered with copper wire, without copper wire and tail), and there was no significant difference in the rates of adhesion to the different parts (p=.7771). The vaginal yeasts showed a high capacity to produce biofilm. CONCLUSIONS: Two vaginal yeasts evaluated showed a high capacity to produce biofilm on IUD. It was confirmed that all parts of the IUD allow the adherence of yeasts. The adherence of C. albicans to different parts of the IUD and its formation of biofilm seems to be important attributes influencing the occurrence of VVC and recurrent VVC. PMID- 18402853 TI - Male condoms that break in use do so mostly by a "blunt puncture" mechanism. AB - BACKGROUND: Published condom breakage studies typically report the percentage of failures but rarely provide any evidence on the mechanism of failure. METHODS: Over a period of 7 years, broken condoms returned to a supplier (SSL, Durex) via consumer complaints were examined to determine the cause of failure. Also, some consumers who reported breakage but did not return condoms were sent a questionnaire on the causes of breakage. Finally, theories proposed for the mechanism of breakage were investigated on a laboratory coital model. RESULTS: Nearly 1000 (n=972) returned condoms made from natural rubber and polyurethane were examined. Visible features on those that were broken, were classified. Evidence combined from examining returns, questionnaire responses and the coital model strongly suggests a single predominant mechanism of failure we named "blunt puncture," where the tip of the thrusting male penis progressively stretches one part of the intact condom wall until it ultimately breaks. CONCLUSIONS: Blunt puncture appears to be the mechanism of breakage responsible for more than 90% of condom breakage not attributable to misuse. Knowledge of the main mechanism of breakage should help develop better user instructions, better test methods and, ultimately, better condoms. PMID- 18402854 TI - Hormonal contraception and the risks of STI acquisition: results of a feasibility study to plan a future randomized trial. AB - BACKGROUND: Because of limitations in observational studies, a randomized controlled trial (RCT) would help clarify whether hormonal contraception increases the risks of acquiring a sexually transmitted infection (STI). However, the feasibility of such a trial is uncertain. STUDY DESIGN: We conducted a study to assess the feasibility of conducting a RCT that would compare the acquisition risk for Chlamydia trachomatis and Neisseria gonorrhoeae in women randomized to an intrauterine device (IUD) or depot medroxyprogesterone acetate (DMPA). In our cross-sectional survey conducted at three clinics, we gave information on a potential RCT to clients, asked them questions to assess comprehensibility and finally asked respondents whether they would consider enrolling in such a trial. In addition, the 190 participants provided urine or endocervical swab specimens so we could estimate the prevalence of STIs. RESULTS: Overall, 70% of participants stated that they would take part in a future trial and accept randomization to either the IUD or DMPA. Participant understanding of the trial requirements was high. Twenty-nine percent of the participants were infected with either N. gonorrhoeae or C. trachomatis. CONCLUSION: With a high prevalence of STI in this population and the apparent willingness of appropriate candidates to participate, an RCT to measure risks of incident STI infection from hormonal contraception appears feasible. PMID- 18402855 TI - Hormonal contraception and HIV prevalence in four African countries. AB - BACKGROUND: The HIV seroprevalence among women aged 15-24 years was compared according to their pattern of contraceptive use in four African countries: Kenya, Lesotho, Malawi and Zimbabwe. STUDY DESIGN: Data were derived from Demographic and Health Surveys (DHS) conducted between 2003 and 2006 on representative samples, totaling 4549 women. RESULTS: It is indicated that users of depo medroxyprogesterone acetate (DMPA) have a significantly higher seroprevalence than nonusers [odds ratio (OR)=1.82, 95% CI=1.63-2.03] and higher than users of oral contraceptives and users of traditional methods. The results were confirmed in a multivariate analysis including as controls, age, duration since first intercourse, urban residence, education, number of sexual partners in the last 12 months and marital status. A somewhat smaller net effect (OR=1.34, 95% CI=1.10 1.63) was found. In contrast, oral contraceptives and traditional methods did not show any risk for HIV (OR=0.96 and 0.92, respectively). CONCLUSION: The increased risk of DMPA was present in three of the four countries investigated, and significant in Zimbabwe and Lesotho, the countries with the highest HIV seroprevalence. The HIV risk attributable to DMPA remained small altogether and was estimated as 6% in the four countries combined. PMID- 18402856 TI - Change in trend of contraceptive uptake--effect of educational leaflets and counseling. AB - BACKGROUND: The study was conducted to determine the impact of counseling and educational leaflets on contraceptive practices of couples. STUDY DESIGN: Randomization of 600 women was done in two groups matched for age, parity and socioeconomic status at the Department of Obstetrics and Gynaecology, Shifa Foundation Community Health Centre, Shifa International Hospital, Islamabad, Pakistan. In Group A, the intervention group was exposed to contraceptive counseling and educational leaflets in the postnatal ward after delivery, whereas in Group B, the nonintervention group was not given any formal contraceptive advice. Later on, both groups were assessed regarding their contraceptive practices. RESULTS: At their follow-up visit (8-12 weeks) postpartum, 19 (6.3%) women in the nonintervention group had started contraceptive use, whereas 153 (50.8%) had decided to start contraception in the next 6 months, and 129 (42.8%) women were still undecided. The main contraceptive user was the male partner (n=117, 38.8%), and the most common method used was coitus interruptus (n=62, 36.3%). In the intervention group, 170 women (56.9%) had started using contraceptives, whereas 129 (43.1%) had decided to start contraceptive use in the next 6 months. The predominant contraceptive user was the females (n=212-70.9%), and the most popular method chosen was oral contraceptive pills (n=111, 37.1%). CONCLUSION: There is a definite increase in contraceptive uptake in women provided with educational leaflets and counseling session with a shift toward use of more reliable contraceptive methods. PMID- 18402857 TI - Comparison of ropivacaine and lidocaine for paracervical block during surgical abortion. AB - BACKGROUND: The study was conducted to compare the effectiveness of ropivacaine and lidocaine as paracervical analgesia for elective abortion by vacuum aspiration. STUDY DESIGN: This single-center double-blinded randomized study tested two different agents for paracervical analgesia in elective abortions: lidocaine (n=57) or ropivacaine (n=57). We assessed intra- and postoperative pain according to both a visual analogical scale rated from 0 to 10 and postoperative analgesic requirements. RESULTS: Mean intraoperative pain was significantly lower in the ropivacaine group (5.23+/-2.72 vs. 4.18+/-2.77, p=.048.). There was no significant difference in mean assessed pain at the end of the intervention or at 2 or 4 h afterward. The rate of subjects requiring additional postoperative analgesia did not differ significantly between the lidocaine and ropivacaine groups [8/59 (13.5%) vs. 6/59 (10.1%), p=.33]. DISCUSSION: Intraoperative pain appears to be less with ropivacaine than with lidocaine. Nonetheless, the clinical difference was slight, as was therefore the benefit of using ropivacaine for paracervical block in elective abortions. PMID- 18402858 TI - Bleeding patterns associated with oral contraceptive use: a review of the literature. PMID- 18402859 TI - Counseling tools alone do not improve method continuation. PMID- 18402861 TI - Hypoxemia: a quick reference. AB - The article describes the calculation of the alveolar-arterial gas gradient and its clinical application in determining the cause of hypoxemia. It also outlines the analysis of arterial blood gases and the clinical approach toward diagnosis and treatment of respiratory disease. PMID- 18402862 TI - Respiratory alkalosis: a quick reference. AB - This article serves as a quick reference for respiratory alkalosis. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402863 TI - Respiratory acidosis: a quick reference. AB - This article serves as a quick reference for respiratory acidosis. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402864 TI - Metabolic alkalosis: a quick reference. AB - This article serves as a quick reference for metabolic alkalosis. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402865 TI - Metabolic acidosis: a quick reference. AB - This article serves as a quick reference for metabolic acidosis. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402866 TI - Anion gap and strong ion gap: a quick reference. AB - This article serves as a quick reference for the anion gap and strong ion gap. Guidelines for analysis and interpretation of changes are presented. PMID- 18402867 TI - Hypercalcemia: a quick reference. AB - This article serves as a quick reference for hypercalcemia. Guidelines for causes, clinical signs, analysis, and diagnosis are presented in a stepwise approach. PMID- 18402868 TI - Hypocalcemia: a quick reference. AB - This article serves as a quick reference for hypocalcemia. Guidelines for causes, clinical signs, and diagnosis are presented in a stepwise approach. PMID- 18402869 TI - Chloride: a quick reference. AB - This article serves as a quick reference for the analysis of chloride abnormalities. Guidelines for causes, signs, and stepwise approaches for corrected hypochloremia and hyperchloremia are presented. PMID- 18402870 TI - Magnesium: a quick reference. AB - This article serves as a quick reference for the distribution, handling, and supplementation of magnesium. It also lists the manifestations and causes of magnesium deficit and provides criteria for the diagnosis of a magnesium deficit. PMID- 18402871 TI - Phosphorus: a quick reference. AB - This article gives a brief overview of phosphorus. The author addresses the main functions of phosphorus in the body, the causes and manifestations of hypophosphatemia and hyperphosphatemia, and suggests further readings. PMID- 18402872 TI - Hyperkalemia: a quick reference. AB - This article serves as a quick reference for hyperkalemia. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402873 TI - Hypokalemia: a quick reference. AB - This article serves as a quick reference for hypokalemia. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402874 TI - Hypernatremia: a quick reference. AB - This article serves as a quick reference for hypernatremia. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402875 TI - Hyponatremia: a quick reference. AB - This article serves as a quick reference for hyponatremia. Guidelines for analysis and causes, signs, and a stepwise approach are presented. PMID- 18402876 TI - Calcium: total or ionized? AB - Measurement of serum total calcium (tCa) has been relied on for assessment of calcium status, despite the fact that it is the ionized calcium (iCa) fraction that has biologic activity. Serum tCa does not accurately predict iCa status in many clinical conditions. For accurate assessment of iCa status, iCa should be directly measured. Anaerobic measurement of serum iCa under controlled conditions provides the most reliable assessment of calcium status; aerobic measurement of iCa with species-specific pH correction is highly correlated with anaerobic measurements. PMID- 18402877 TI - Urinary electrolytes, solutes, and osmolality. AB - Urine chemical analysis can extend "beyond the dipstick" with an understanding of renal physiology and expected changes in electrolyte and solute handling. Urine electrolytes, such as sodium and chloride, can be helpful in discerning prerenal azotemia from acute renal tubular damage, which occur secondary to nephrotoxins or ischemia. Urine osmolality also is essential in determining appropriate antidiuretic hormone action and renal water handling. Urine solutes, such as albumin and brush border enzymes, may be more sensitive than plasma markers for early renal dysfunction. This article reviews these topics and the use of "extended" urine indices in veterinary medicine. PMID- 18402878 TI - Therapeutic approach to electrolyte emergencies. AB - Hypokalemia, hyperkalemia, hyponatremia, hypernatremia, hypocalcemia, and hypercalcemia are commonly seen in emergency medicine. Severe abnormalities in any of these electrolytes can cause potentially life-threatening consequences to the patient. It is essential that the clinician understand and correct (if possible) the underlying cause of each disorder and recognize the importance of the rates of correction, especially with serum sodium disorders. The recommended doses in this article might have to be adjusted to the individual patient, and these modifications must be adjusted again to the pathophysiology of the primary underlying disorder. PMID- 18402879 TI - Therapeutic approach to chronic electrolyte disorders. AB - Chronic disorders of sodium and potassium occur and occasionally need symptomatic therapy. Hypernatremia primarily indicates loss of free water, whereas hyponatremia may be attributable to various problems. It is important not to correct major aberrations of serum sodium concentrations too quickly lest the therapy be more detrimental than the electrolyte abnormality. In distinction, hypokalemia and hyperkalemia may be corrected quickly. Hypomagnesemia is relatively common, but its clinical significance is still being determined. PMID- 18402880 TI - Making sense of blood gas results. AB - Recent technologic advances have allowed the production and marketing of cage side blood gas analyzers to private practitioners. The widespread use of cage side portable blood gas analyzers in veterinary practices has increased the need to develop the basic skills of blood gas analysis as part of a tool kit for practicing veterinarians. Rapid expansion of emergency and critical care medicine as a specialty and increased numbers of veterinary emergency and veterinary specialty practices have occurred concurrently with the availability of blood gas analyzers that are affordable for private practitioners. As a result, evaluation of blood gas results is no longer an activity confined to academic institutions and has become a daily part of many practicing veterinarians' activities. PMID- 18402881 TI - Metabolic acid-base disorders in the critical care unit. AB - The recognition and management of acid-base disorders is a commonplace activity in the critical care unit, and the role of weak and strong acids in the genesis of metabolic acid-base disorders is reviewed. The clinical approach to patients with metabolic alkalosis and metabolic acidosis is discussed in this article. PMID- 18402882 TI - Fluid therapy: options and rational administration. AB - Fluid administration is a primary component of therapy in many small animal patients. Several different classes of fluid may be given, and there are multiple options within each class. The type, route, volume, and rate of fluid administered should be tailored to the patient's signalment, disease or injury state, and response to the administration of fluids. Monitoring vital parameters and bedside monitoring of laboratory variables allow assessment of fluid therapy success or failure. Successful fluid administration also requires that parenteral fluid therapy ultimately be discontinued with minimal adverse effects for the patient. PMID- 18402883 TI - Colloids: current recommendations. AB - Colloids are increasingly becoming considered indispensable in the management of critically ill patients. Typical indications for colloid administration include patients with tissue edema, hypovolemia, and low oncotic pressure. Current guidelines for the use of colloids in veterinary patients balance the purported benefits of colloid fluid administration with the potential risks, such as volume overload and coagulation disturbances. This article focuses primarily on hydroxyethyl starches, because they are the most commonly used colloid in veterinary practice, and because recent advances in colloid therapy have been achieved with this colloid. Newer colloids have been modified to limit effects on the coagulation system, and they may be used to modulate the inflammatory response, which could prove to be particularly useful in the management of critically ill patients. A better understanding of how different fluids influence the host response may enable us to explore new applications of fluid replacement therapy beyond simply replenishing volume deficits. PMID- 18402884 TI - The therapeutic use of 25% human serum albumin in critically ill dogs and cats. AB - Twenty-five percent human serum albumin (HSA) is a foreign protein and can potentially cause immune-mediated reactions. For this reason, the author only recommends 25% HSA use after risk analysis shows that the benefits outweigh the potential risks of adverse events. If it is apparent that a critically ill animal may succumb to its illness because of the problems associated with severe hypoalbuminemia, the benefit outweighs the risk. The veterinarian must inform the owner of potential delayed immune-mediated reactions, describe these lesions, and follow the case weekly to ensure that no reaction has occurred. Although there are many positive attributes to the administration of 25% HSA, there seems to be specific situations in which 25% HSA may be indicated and others in which it may not be indicated. PMID- 18402885 TI - Complications of fluid therapy. AB - The intravenous administration of fluids is one of the most important aspects of patient care in hospitalized animals. Intravenous fluids are administered to replace or prevent dehydration, treat hypovolemic shock and intravascular volume depletion, correct acid-base and electrolyte abnormalities, and maintain vascular access for administration of drugs, blood product components, and parenteral nutrition. Intravenous catheterization also can provide a means of blood sample collection, thus avoiding frequent and uncomfortable venipunctures in critically ill animals. Although the benefits of intravenous catheterization and fluid administration are numerous, inherent risks are associated with the procedures, and care must be taken to avoid potential complications. PMID- 18402886 TI - Pediatric fluid therapy. AB - Many conditions of pediatric patients require fluid therapy. Depending on the veterinarian's assessment of hydration and perfusion status, fluids can be administered orally, subcutaneously, intraperitoneally, intravenously, or by the intraosseous route. Pediatric patients are prone to hypothermia, hypovolemia, hypoglycemia, and hypokalemia, which must be addressed during fluid therapy in pediatric patients. Typical parameters used to assess hydration status in adult animals do not always apply to pediatric patients. Veterinarians should be aware of differences between pediatric patients and adult animals in terms of physical assessment, common presentations, and fluid requirements for resuscitation and maintenance needs. PMID- 18402887 TI - Assessment and treatment of hypovolemic states. AB - Hypovolemia and hypoperfusion are common life-threatening problems in animals presented to the emergency veterinarian. Assessment of physical findings and rapid recognition and treatment of abnormal tissue perfusion are crucial in optimizing outcome. The clinician should be familiar with the disease being treated and the types of fluids that are available. Development of a fluid therapy plan to correct life-threatening abnormalities and patient monitoring during treatment play an important role in patient outcome. PMID- 18402888 TI - Fluid resuscitation and the trauma patient. AB - Traumatic shock can result from one or more of the following: hypovolemia, increased capillary permeability and vasodilatation, impaired myocardial contractility or dysfunction, blood loss, and cardiovascular obstruction. Animals with traumatic injuries are less tolerant of sudden increases in hydrostatic pressure, which can exacerbate fluid leakage into damaged tissues and disrupt blood clots. Pain amplifies the shock response, and analgesic therapy is always recommended in the trauma patient. Therapy must be individualized and monitored closely. PMID- 18402889 TI - Fluid therapy in vomiting and diarrhea. AB - Fluid therapy in the patient with vomiting and diarrhea is essential to correct hypovolemia, dehydration, acid-base imbalance, and serum electrolyte abnormalities. Prediction of acid-base or electrolyte disturbances is difficult; therefore, point of care testing is beneficial to optimize therapy. This article focuses on the pathophysiology and treatment of hypovolemia, dehydration, electrolyte disturbances, and acid-base derangements resulting from and associated with vomiting and diarrhea. PMID- 18402890 TI - Managing fluid and electrolyte disorders in renal failure. AB - Because of the role of the kidneys in maintaining homeostasis in the body, kidney failure leads to derangements of fluid, electrolyte, and acid-base balance. The most effective therapy of a uremic crisis is careful management of fluid balance, which involves thoughtful assessment of hydration, a fluid treatment plan personalized for the specific patient, repeated and frequent reassessment of fluid and electrolyte balance, and appropriate changes in the treatment plan in response to the rapidly changing clinical situation of the patient that has renal failure. Disorders of sodium, chloride, potassium, calcium, and phosphorus are commonly encountered in renal failure and may be life threatening. Treatment of metabolic acidosis and nutritional support are frequently needed. PMID- 18402891 TI - Fluid and electrolyte therapy in endocrine disorders: diabetes mellitus and hypoadrenocorticism. AB - Diabetic ketoacidosis, hyperglycemic hyperosmolar syndrome, and acute adrenal crisis are life-threatening endocrine emergencies that have marked effects on fluid, electrolyte, and acid-base homeostasis. Rapid identification of these disorders and aggressive therapy to correct fluid, electrolyte, and acid-base imbalances are crucial to a successful outcome for the patient. An understanding of the pathophysiology behind the development of these endocrine disorders helps to guide therapy and improves the clinical outcome. PMID- 18402892 TI - Fluid therapy in patients with pulmonary disease. AB - Fluid therapy in patients with pulmonary disease is challenging. Although a single set of rules cannot be applied to every patient, the following guidelines can be used when managing patients with pulmonary disease. Euvolemic patients with adequate tissue perfusion should be given sufficient isotonic fluid to balance insensible losses. If severe pulmonary compromise is present, cessation of all fluid therapy may be considered if the patient is able to match its losses by voluntary intake. In hypovolemic or hypotensive patients, small boluses of isotonic crystalloids or colloids should be given to restore perfusion, avoiding rates of more than 30 mL/kg an hour for isotonic crystalloids. If perfusion is not restored by adequate volume resuscitation, vasopressors or positive inotropes should be administered to prevent fluid overload and deterioration in pulmonary function. PMID- 18402893 TI - Maintaining fluid and electrolyte balance in heart failure. AB - Advanced heart failure and its treatment are often associated with a variety of hemodynamic, fluid, and electrolyte derangements. This article gives the practitioner an overview of the pathophysiology of common fluid and electrolyte alterations present in animals with heart failure, highlighting specific clinical correlates. Additionally, specific therapeutic interventions are discussed to manage these fluid and electrolyte abnormalities. PMID- 18402894 TI - Contrast-induced acute kidney injury. AB - Cardiac angiography and coronary/vascular interventions depend on iodinated contrast media and consequently pose the risk of contrast-induced acute kidney injury (AKI). This is an important complication that accounts for a significant number of cases of hospital-acquired renal failure, with adverse effects on prognosis and health care costs. The epidemiology and pathogenesis of contrast induced AKI, baseline renal function measurement, risk assessment, identification of high-risk patients, contrast medium use, and preventive strategies are discussed in this report. An advanced algorithm is suggested for the risk stratification and management of contrast-induced AKI as it relates to patients undergoing cardiovascular procedures. Contrast-induced AKI is likely to remain a significant challenge for cardiologists in the future because the patient population is aging and chronic kidney disease and diabetes are becoming more common. PMID- 18402895 TI - Stem cell mobilization by granulocyte colony-stimulating factor for myocardial recovery after acute myocardial infarction: a meta-analysis. AB - OBJECTIVES: The objective of this meta-analysis was to evaluate the effect of stem cell mobilization by granulocyte colony-stimulating factor (G-CSF) on myocardial regeneration on the basis of a synthesis of the data generated by randomized, controlled clinical trials of G-CSF after acute myocardial infarction (AMI). BACKGROUND: Experimental studies and early-phase clinical trials suggest that stem cell mobilization by G-CSF may have a positive impact on cardiac regeneration after AMI. The role of G-CSF in patients with AMI remains unclear considering the inconsistent results of several clinical trials. METHODS: For our analysis, PubMed, the Cochrane Central Register of Controlled Trials, conference proceedings from major cardiology meetings, and Internet-based sources of information on clinical trials in cardiology from January 2003 to August 2007 served as sources. Two reviewers independently identified studies and abstracted data on sample size, baseline characteristics, and outcomes of interest. Eligible studies were randomized trials with stem cell mobilization by G-CSF after reperfused AMI that reported data regarding the change in left ventricular ejection fraction (LVEF) at follow-up. RESULTS: Ten trials using stem cell mobilization by G-CSF, including 445 patients, met the inclusion criteria. Significant improvement in LVEF at follow-up was observed in both the G-CSF and placebo groups. Compared with placebo, stem cell mobilization by G-CSF did not enhance the improvement of LVEF at follow-up (mean difference 1.32% [95% confidence interval -1.52 to 4.16; p = 0.36]). Moreover, the mean difference of reduction of infarct size between the treatment and placebo groups was -0.15 (95% confidence interval -0.38 to 0.07, p = 0.17). CONCLUSIONS: Cumulatively, available evidence does not support a beneficial effect of G-CSF in patients with AMI after reperfusion. PMID- 18402896 TI - An imperfect syllogism: granulocyte colony-stimulating factor mobilization and cardiac regeneration. PMID- 18402897 TI - Lipid levels after acute coronary syndromes. AB - OBJECTIVES: This analysis from the LUNAR (Limiting UNdertreatment of lipids in ACS with Rosuvastatin) study assessed lipid changes 1 to 4 days after onset of acute coronary syndromes (ACS), before initiation of study treatment. BACKGROUND: Early studies indicated that cholesterol levels decrease significantly after ACS. However, most studies were small or did not measure low-density lipoprotein cholesterol (LDL-C) directly, and many used nonfasting or retrospective data. More recent studies suggest less pronounced changes in cholesterol levels after ACS. METHODS: The LUNAR trial is a prospective, multicenter, randomized, open label study in adults hospitalized for acute ST-segment elevation myocardial infarction (STEMI), non-STEMI, or unstable angina (UA). Blood samples were taken at median times after onset of ACS symptoms of 26 h (Day 1, fasting or nonfasting sample), 43 h (Day 2, fasting sample), and 84 h (Day 4, fasting sample) for direct measurement of serum lipid levels before study treatments were started. RESULTS: Of 507 patients available for analysis, 212 were admitted for STEMI, 176 for non-STEMI, and 119 for UA. The LDL-C levels decreased in the 24 h after admission (from 136.2 to 133.5 mg/dl), followed by an increase over the subsequent 2 days (to 141.8 mg/dl). These changes did not seem to be clinically meaningful. Similar changes were observed for total cholesterol and smaller changes for high-density lipoprotein cholesterol; fasting triglyceride levels did not change. CONCLUSIONS: Mean lipid levels vary relatively little in the 4 days after an ACS and can be used to guide selection of lipid-lowering medication. PMID- 18402898 TI - Lipid levels in the post-acute coronary syndrome setting: destabilizing another myth? PMID- 18402899 TI - Intensive lipid lowering with atorvastatin in patients with coronary heart disease and chronic kidney disease: the TNT (Treating to New Targets) study. AB - OBJECTIVES: This subanalysis of the TNT (Treating to New Targets) study investigates the effects of intensive lipid lowering with atorvastatin in patients with coronary heart disease (CHD) with and without pre-existing chronic kidney disease (CKD). BACKGROUND: Cardiovascular disease is a major cause of morbidity and mortality in patients with CKD. METHODS: A total of 10,001 patients with CHD were randomized to double-blind therapy with atorvastatin 80 mg/day or 10 mg/day. Patients with CKD were identified at baseline on the basis of an estimated glomerular filtration rate (eGFR) <60 ml/min/1.73 m(2) using the Modification of Diet in Renal Disease equation. The primary efficacy outcome was time to first major cardiovascular event. RESULTS: Of 9,656 patients with complete renal data, 3,107 had CKD at baseline and demonstrated greater cardiovascular comorbidity than those with normal eGFR (n = 6,549). After a median follow-up of 5.0 years, 351 patients with CKD (11.3%) experienced a major cardiovascular event, compared with 561 patients with normal eGFR (8.6%) (hazard ratio [HR] = 1.35; 95% confidence interval [CI] 1.18 to 1.54; p < 0.0001). Compared with atorvastatin 10 mg, atorvastatin 80 mg reduced the relative risk of major cardiovascular events by 32% in patients with CKD (HR = 0.68; 95% CI 0.55 to 0.84; p = 0.0003) and 15% in patients with normal eGFR (HR = 0.85; 95% CI 0.72 to 1.00; p = 0.049). Both doses of atorvastatin were well tolerated in patients with CKD. CONCLUSIONS: Aggressive lipid lowering with atorvastatin 80 mg was both safe and effective in reducing the excess of cardiovascular events in a high-risk population with CKD and CHD. PMID- 18402900 TI - A randomized comparison of triple-site versus dual-site ventricular stimulation in patients with congestive heart failure. AB - OBJECTIVES: We compared the effects of triple-site versus dual-site biventricular stimulation in candidates for cardiac resynchronization therapy. BACKGROUND: Conventional biventricular stimulation with a single right ventricular (RV) and a single left ventricular (LV) lead is associated with persistence of cardiac dyssynchrony in up to 30% of patients. METHODS: This multicenter, single-blind, crossover study enrolled 40 patients (mean age 70 +/- 9 years) with moderate-to severe heart failure despite optimal drug treatment, a mean LV ejection fraction of 26 +/- 11%, and permanent atrial fibrillation requiring cardiac pacing for slow ventricular rate. A cardiac resynchronization therapy device connected to 1 RV and 2 LV leads, inserted in 2 separate coronary sinus tributaries, was successfully implanted in 34 patients. After 3 months of biventricular stimulation, the patients were randomly assigned to stimulation for 3 months with either 1 RV and 2 LV leads (3-V) or to conventional stimulation with 1 RV and 1 LV lead (2-V), then crossed over for 3 months to the alternate configuration. The primary study end point was quality of ventricular resynchronization (Z ratio). Secondary end points included reverse LV remodeling, quality of life, distance covered during 6-min hall walk, and procedure-related morbidity and mortality. Data from the 6- and 9-month visits were combined to compare end points associated with 2-V versus 3-V. RESULTS: Data eligible for protocol-defined analyses were available in 26 patients. No significant difference in Z ratio, quality of life, and 6-min hall walk was observed between 2-V and 3-V. However, a significantly higher LV ejection fraction (27 +/- 11% vs. 35 +/- 11%; p = 0.001) and smaller LV end-systolic volume (157 +/- 69 cm(3) vs. 134 +/- 75 cm(3); p = 0.02) and diameter (57 +/- 12 mm vs. 54 +/- 10 mm; p = 0.02) were observed with 3 V than with 2-V. There was a single minor procedure-related complication. CONCLUSIONS: Cardiac resynchronization therapy with 1 RV and 2 LV leads was safe and associated with significantly more LV reverse remodeling than conventional biventricular stimulation. PMID- 18402901 TI - Cardiac resynchronization therapy: the more pacing sites, the better the outcome? PMID- 18402902 TI - Aortic valve replacement for low-flow/low-gradient aortic stenosis operative risk stratification and long-term outcome: a European multicenter study. AB - OBJECTIVES: We evaluated a large multicenter series of patients operated on for low-flow/low-gradient aortic stenosis (LF/LGAS) to stratify the operative risk, assess whether perioperative mortality has decreased over recent years, and analyze the post-operative outcome. BACKGROUND: Although LF/LGAS is classically associated with a high operative risk, few data are available concerning the results of surgery in this setting. METHODS: A total of 217 consecutive patients (168 men, 77%) with severe aortic stenosis (area <1 cm(2)), low ejection fraction (EF) ( or =30 kg/m(2)). CONCLUSION: Quality of dietary carbohydrates does not appear to be associated with serum hs CRP levels. Among obese individuals, higher dietary GL appears to be related to lower hs-CRP levels. Due to the limited number of studies on this topic and their conflicting results, further investigation is warranted. PMID- 18402915 TI - Special issue: Novel RNA nucleotidyl transferases and gene regulation. Preface. PMID- 18402923 TI - Isoenzymatic variability of Leishmania infantum in Tunisia concerning 254 human strains. AB - The different clinical forms of leishmaniasis are the result of both the immunological status of individuals and the species of the parasite causing the infection. In Mediterranean countries, the Leishmania infantum complex groups zymodemes which are responsible for visceral, cutaneous and exceptionally cutaneomucosal or mucosal leishmaniasis. We report in this study a synthesis concerning 254 cases of L. infantum that have been characterized at the "Laboratoire de Parasitologie" of the Rabta Hospital. The strains were isolated from human cases of visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) by culture on NNN medium: 156 VL cases and 98 CL cases. The isoenzymatic characterization revealed three zymodemes of L. infantum. * L. infantum MON 1, a common zymodeme of VL,occurred in 154 cases (61%): 147 VL (95%) and 7 CL (5%). All CL cases were from the northern provinces, six of them occurring during an epidemic disease in 2001. * L. infantum MON 24, a common zymodeme of CL in the north, occurred in 98 cases (38.5%): 91 CL (93%) and 7 VL (7%). The seven VL cases were immunocompetent children aged from 8 months to 9 years and native of northern Tunisia. Two of the CL cases were from central regions of the country. This is the first time that cases from these regions are reported. * L. infantum MON 80, an uncommon zymodeme in Tunisia, occurred in two VL cases (0.5%): two children aged 7 and 5. The small number of strains of this zymodeme does not allow understanding of its epidemiological role. The results of this study indicate a low enzymatic variability of L. infantum in the country. However, our study includes only human strains and should be extended to animal ones (dogs, rodents and sand flies). This would lead to a better understanding of the epidemiology of leishmaniasis in Tunisia. PMID- 18402924 TI - [Non-valvular cardiac devices endocarditis]. AB - The risk of infective endocarditis on pacemaker or ICD is not negligible and has increased in recent years. Several host-related, procedure-related, or device related risk factors have been recognized. Owing to its potential severity, the possibility of infective endocarditis should be envisaged in patients with repeated pulmonary infections or documented bacteremia and transesophageal echocardiography should then be used. The most common germs causing pacemaker endocarditis are staphylococci. Treatment requires prolonged antibiotic therapy and retrieval of the pacemaker and leads. PMID- 18402925 TI - [Prophylaxis of infective endocarditis: trends and new recommendations]. AB - The rationale having led to the profound change in the guidelines on the prevention of endocarditis is presented. The current trend to abandoning systematic antibiotic prophylaxis in patients at risk for infective endocarditis is in fact based upon a sound scientific background. Systematic antibiotic prophylaxis remains necessary, however, in the category of patients at very high risk. PMID- 18402926 TI - [Is there a change in the epidemiologic profile of infective endocarditis?]. PMID- 18402927 TI - [Timing and indications for surgical intervention in infective endocarditis]. AB - This paper reviews current knowledge on the indications for and timing of cardiac surgery in patients with infective endocarditis. The main indications for surgery are haemodynamic compromise, persisting infection, peripheral embolisation, large size of vegetations, large valvular and paravalvular damage and infections caused by certain microorganisms. PMID- 18402928 TI - Editors' comments on this special issue devoted to macroergonomics. PMID- 18402930 TI - PubMedAlertMe--standalone Windows-based PubMed SDI software application. AB - PubMedAlertMe is a Windows-based software system for automatically receiving e mail alert messages about recent publications listed on PubMed. The e-mail messages contain links to newly available abstracts listed on PubMed describing publications that were selectively returned from a specified list of queries. Links are also provided to directly export citations to EndNote, and links are provided to directly forward articles to colleagues. The program is standalone. Thus, it does not require a remote mail server or user registration. PubMedAlertMe is free software, and can be downloaded from: http://amp.pharm.mssm.edu/PubMedAlertMe/PubMedAlertMe_setup.zip. PMID- 18402929 TI - Aromatase distribution in the monkey temporal neocortex and hippocampus. AB - Numerous studies have shown that neuronal plasticity in the hippocampus and neocortex is regulated by estrogen and that aromatase, the key enzyme for estrogen biosynthesis, is present in cerebral cortex. Although the expression pattern of aromatase mRNA has been described in the monkey brain, its precise cellular distribution has not been determined. In addition, the degree to which neuronal aromatase is affected by gonadal estrogen has not been investigated. In this study, we examined the immunohistochemical distribution of aromatase in young ovariectomized female rhesus monkeys with or without long-term cyclic estradiol treatment. Both experimental groups showed that aromatase is localized in a large population of CA1-3 pyramidal cells, in granule cells of the dentate gyrus and in some interneurons in which it was co-expressed with the calcium binding proteins calbindin, calretinin, and parvalbumin. Moreover, numerous pyramidal cells were immunoreactive for aromatase in the neocortex, whereas only small subpopulations of neocortical interneurons were immunoreactive for aromatase. The widespread expression of the protein in a large neuronal population suggests that local intraneuroral estrogen synthesis may contribute to estrogen-induced synaptic plasticity in monkey hippocampus and neocortex of female rhesus monkeys. In addition, the apparent absence of obvious differences in aromatase distribution between the two experimental groups suggests that these localization patterns are not dependent on plasma estradiol levels. PMID- 18402931 TI - Automatic segmentation of age-related macular degeneration in retinal fundus images. AB - Every year an increasing number of people are affected by age-related macular degeneration (ARMD). Consequently, vast amount of information is accumulated in medical databases and manual classification of this information is becoming more and more difficult. Therefore, there is an increasing interest in developing automated evaluation methods to follow up the diseases. In this paper, we have presented an automatic method for segmenting the ARMD in retinal fundus images. Previously used direct segmentation techniques, generating unsatisfactory results in some cases, are more complex and costly than our inverse method. This is because of the fact that the texture of unhealthy areas of macula is quite irregular and varies from eye to eye. Therefore, a simple inverse segmentation method is proposed to exploit the homogeneity of healthy areas of the macula rather than unhealthy areas. This method first extracts healthy areas of the macula by employing a simple region growing method. Then, blood vessels are also extracted and classified as healthy regions. In order to produce the final segmented image, the inverse image of the segmented image is generated as unhealthy region of the macula. The performance of the method is examined on various qualities of retinal fundus images. The segmentation method without any user involvement provides over 90% segmentation accuracy. Segmented images with reference invariants are also compared with consecutive images of the same patient to follow up the changes in the disease. PMID- 18402932 TI - DIF-1 induces the basal disc of the Dictyostelium fruiting body. AB - The polyketide DIF-1 induces Dictyostelium amoebae to form stalk cells in culture. To better define its role in normal development, we examined the phenotype of a mutant blocking the first step of DIF-1 synthesis, which lacks both DIF-1 and its biosynthetic intermediate, dM-DIF-1 (des-methyl-DIF-1). Slugs of this polyketide synthase mutant (stlB(-)) are long and thin and rapidly break up, leaving an immotile prespore mass. They have approximately 30% fewer prestalk cells than their wild-type parent and lack a subset of anterior-like cells, which later form the outer basal disc. This structure is missing from the fruiting body, which perhaps in consequence initiates culmination along the substratum. The lower cup is rudimentary at best and the spore mass, lacking support, slips down the stalk. The dmtA(-) methyltransferase mutant, blocked in the last step of DIF-1 synthesis, resembles the stlB(-) mutant but has delayed tip formation and fewer prestalk-O cells. This difference may be due to accumulation of dM-DIF-1 in the dmtA(-) mutant, since dM-DIF-1 inhibits prestalk-O differentiation. Thus, DIF 1 is required for slug migration and specifies the anterior-like cells forming the basal disc and much of the lower cup; significantly the DIF-1 biosynthetic pathway may supply a second signal - dM-DIF-1. PMID- 18402933 TI - Identification of cis-regulatory elements from the C. elegans T-box gene mab-9 reveals a novel role for mab-9 in hypodermal function. AB - We have identified Conserved Non-coding Elements (CNEs) in the regulatory region of Caenorhabditis elegans and Caenorhabditis briggsae mab-9, a T-box gene known to be important for cell fate specification in the developing C. elegans hindgut. Two adjacent CNEs (a region 78 bp in length) are both necessary and sufficient to drive reporter gene expression in posterior hypodermal cells. The failure of a genomic mab-9::gfp construct lacking this region to express in posterior hypodermis correlates with the inability of this construct to completely rescue the mab-9 mutant phenotype. Transgenic males carrying this construct in a mab-9 mutant background exhibit tail abnormalities including morphogenetic defects, altered tail autofluorescence and abnormal lectin-binding properties. Hermaphrodites display reduced susceptibility to the C. elegans pathogen Microbacterium nematophilum. This comparative genomics approach has therefore revealed a previously unknown role for mab-9 in hypodermal function and we suggest that MAB-9 is required for the secretion and/or modification of posterior cuticle. PMID- 18402934 TI - Review of fate-mapping studies of osteogenic cranial neural crest in vertebrates. AB - Recent years have witnessed renewed interest in defining the embryonic cell populations that directly contribute to the bony skull. This question lies at the intersection of several important developmental, clinical and evolutionary interests. Until recently, our collective understanding of the embryonic origin of the vertebrate osteocranium has been based on a small number of reports published solely using avian models. As data gradually accumulates from other, distantly related species (e.g., mouse and frog), we can begin to evaluate long standing assumptions regarding the behavior of osteogenic (bone-forming) neural crest cells within a wider phylogenetic and comparative context. In this review, we summarize data collected to date in three major vertebrate taxa: amphibians, birds and mammals. We highlight three largely unexplored topics within the field of osteogenic neural crest development: 1) disagreements in bone tissue origin within and across current model systems; 2) whether the pattern of neural crest cell contribution to skull bone is evolutionarily conservative or labile; and 3) how our understanding of development and morphology will benefit from fate maps using currently unexamined animal models. PMID- 18402935 TI - Insulin, PKC signaling pathways and synaptic remodeling during memory storage and neuronal repair. AB - Protein kinase C (PKC) is involved in synaptic remodeling, induction of protein synthesis, and many other processes important in learning and memory. Activation of neuronal protein kinase C correlates with, and may be essential for, all phases of learning, including acquisition, consolidation, and reconsolidation. Protein kinase C activation is closely tied to hydrolysis of membrane lipids. Phospholipases C and A2 produce 1,2-diacylglycerol and arachidonic acid, which are direct activators of protein kinase C. Phospholipase C also produces inositol triphosphate, which releases calcium from internal stores. Protein kinase C interacts with many of the same pathways as insulin; therefore, it should not be surprising that insulin signaling and protein kinase C activation can both have powerful effects on memory storage and synaptic remodeling. However, investigating the possible roles of insulin in memory storage can be challenging, due to the powerful peripheral effects of insulin on glucose and the low concentration of insulin in the brain. Although peripheral for insulin, synthesized in the beta-cells of the pancreas, is primarily involved in regulating glucose, small amounts of insulin are also present in the brain. The functions of this brain insulin are inadequately understood. Protein kinase C may also contribute to insulin resistance by phosphorylating the insulin receptor substrates required for insulin signaling. Insulin is also responsible insulin long term depression, a type of synaptic plasticity that is also dependent on protein kinase C. However, insulin can also activate PKC signaling pathways via PLC gamma, Erk 1/2 MAP kinase, and src stimulation. Taken together, the available evidence suggests that the major impact of protein kinase C and its interaction with insulin in the mature, fully differentiated nervous system appears to be to induce synaptogenesis, enhance memory, reduce Alzheimer's pathophysiology, and stimulate neurorepair. PMID- 18402936 TI - YSK2821, a newly synthesized indoledione derivative, inhibits cell proliferation and cell cycle progression via the cell cycle-related proteins by regulating phosphatidylinositol-3 kinase cascade in vascular smooth muscle cells. AB - Indoledione derivatives have pronounced biological effects, i.e., cytotoxic activities against cancer cell lines and antifungal and antibacterial activities. The present study was designed to investigate the effects of YSK2821, a newly synthesized indoledione derivative, on platelet-derived growth factor (PDGF-BB) induced vascular smooth muscle cell (VSMC) proliferation, as well as the molecular mechanisms of the anti-proliferative effects of YSK2821 in VSMCs. We found that YSK2821 caused the accumulation of cells in the G1 phase of the cell cycle and inhibited [3H]-thymidine incorporation. We demonstrated that YSK2821 remarkably decreased Akt kinase phosphorylation as the mechanism by which YSK2821 suppressed cell signal transduction events in VSMC proliferation. Furthermore, in terms of the effects of YSK2821 on cell cycle-related proteins, YSK2821 enhanced the expression of the cyclin-dependent protein kinase (CDK) inhibitor p27 and down-regulated CDK2 and cyclin E expression, but did not affect CDK4 and cyclin D1 expression. YSK2821 also inhibited the phosphorylation of Rb, a key regulator in the cell cycle. These results indicate that YSK2821, a newly synthesized indoledione derivative, may inhibit VSMC proliferation via a phosphatidylinositol (PI)-3 kinase-dependent pathway, and thus shed light on a novel role for YSK2821 as a potential preventive regulator of cardiovascular disease. PMID- 18402938 TI - A combined behavioral and morphological study on the effects of fetal asphyxia on the nigrostriatal dopaminergic system in adult rats. AB - Fetal asphyxic insults in the brain are known to be associated with developmental neurological problems like neuromotor disorders. However, little is known about the long-term consequences of fetal asphyxia (FA). For that reason, the present study investigated the long-term effects of FA on motor behavior and dopaminergic circuitry. FA was induced at embryonic day 17 by 75-minute clamping of the uterine circulation. SHAM animals underwent the same procedure except for the clamping. This was followed by full-term vaginal delivery of animals in all groups (FA, SHAM and untreated controls). At 6 months, basal and amphetamine induced locomotor activity was measured during open field testing. Brain sections were stained for tyrosine hydroxylase (TH) and glial fibrillary acidic protein (GFAP). TH-positive cells and GFAP-positive cells in substantia nigra pars compacta (SN(C)) and striatum were counted using design-based stereology. Moreover, TH-immunoreactivity in the striatum was assessed by grey value measurements. Behavioral analysis demonstrated that SHAM and FA showed less basal and amphetamine-induced activity than controls. Histochemically, FA decreased the number of TH-positive neurons in the SN(C) and lowered TH-positive in the striatum. Furthermore, more GFAP-positive cells were found in the SN(C) and striatum in FA than in either control and SHAM groups. Additionally, FA animals showed ventriculomegaly associated with smaller white matter as well as grey matter volumes. The data show that FA was associated with deficits in both dopamine-related motor behavior and biochemistry. These alterations were associated with nigrostriatal astrogliosis. The present study demonstrates the sensitivity of the dopaminergic system towards FA. PMID- 18402939 TI - In vivo imaging of seizure activity in a novel developmental seizure model. AB - The immature brain is exceptionally susceptible to seizures. However, it remains unclear whether seizures occurring during development affect critical processes underlying neural circuit formation, leading to long-term functional consequences. Here we characterize a novel in vivo model system of developmental seizures based on the transparent albino Xenopus laevis tadpole, which allows direct examination of seizure activity, and seizure-induced effects on neuronal development within the intact unanesthetized brain. Pentylenetetrazol (PTZ), kainic acid, bicuculline, picrotoxin, 4-aminopyridine, and pilocarpine were tested for their ability to induce behavioral seizures in freely swimming tadpoles when bath applied. All six chemoconvulsants consistently induced similar patterns of abnormal behavior in a dose-dependent manner, characterized by convulsive clonus-like motor patterns with periods of behavioral arrest. Extracellular field recordings demonstrated rhythmic synchronous epileptiform electrographic responses induced by convulsants irrespective of mechanism of action, that could be terminated by the anti-epileptic drug valproate. PTZ induced seizures were further characterized using in vivo two-photon fluorescence imaging of neuronal calcium dynamics, in unanesthetized immobilized tadpoles. Imaging of calcium dynamics during PTZ-induced seizures revealed waves of neural activity propagating through large populations of neurons within the brain. Analysis of single-cell responses demonstrated distinct synchronized high amplitude calcium spikes not observed under baseline conditions. Similar to other developmental seizure models, prolonged seizures failed to induce marked neuronal death within the brain, detected by cellular propidium iodide incorporation in vivo or TUNEL labeling. This novel developmental seizure model system has distinct advantages for controlled seizure induction, and direct visualization of both seizure activity and seizure-induced effects on individual developing neurons within the intact unanesthetized brain. Such a system is necessary to address important questions relating to the long-term impact of common perinatal seizures on developing neural circuits. PMID- 18402937 TI - Anti-apoptotic actions of vasopressin in H32 neurons involve MAP kinase transactivation and Bad phosphorylation. AB - Vasopressin (VP) secreted within the brain modulates neuronal function acting as a neurotransmitter. Based on the observation that VP prevented serum deprivation induced cell death in the neuronal cell line, H32, which expresses endogenous V1 receptors, we tested the hypothesis that VP has anti-apoptotic properties. Flow cytometry experiments showed that 10 nM VP prevented serum deprivation-induced cell death and annexin V binding. Serum deprivation increased caspase-3 activity in a time and serum concentration dependent manner, and VP prevented these effects through interaction with receptors of V1 subtype. The signaling pathways mediating the anti-apoptotic effect of VP involve mitogen activated protein (MAP) kinase and extracellular signal-regulated kinases (ERK), Ca(2+)/calmodulin dependent kinase (CaMK) and protein kinase C (PKC). Western blot analyses revealed time-dependent decreases of Bad phosphorylation and increases in cytosolic levels of cytochrome c following serum deprivation, effects which were prevented by 10 nM VP. These data demonstrate that activation of endogenous V1 VP receptors prevents serum deprivation-induced apoptosis, through phosphorylation inactivation of the pro-apoptotic protein, Bad, and consequent decreases in cytosolic cytochrome c and caspase-3 activation. The data suggest that VP has anti-apoptotic activity in neurons and that VP may act as a neuroprotective agent in the brain. PMID- 18402940 TI - Blastocyst formation, pregnancy, and birth derived from human oocytes cryopreserved for 5 years. AB - OBJECTIVE: To report a live birth after the transfer of a single blastocyst derived from a human oocyte cryopreserved for 5 years. DESIGN: Case report. SETTING: Private assisted reproduction center. PATIENT(S): A 39-year-old woman with tubal infertility and her partner with male-related infertility. INTERVENTION(S): Oocyte cryopreservation (in 1.5 mol/L 1,2-propanediol and 0.3 mol/L sucrose by slow freezing-rapid thawing protocol) when the patient was 34 years old. Oocyte thawing after 5 years of cryostorage. Insemination by intracytoplasmic sperm injection of the three best surviving oocytes, according to the Italian law regulating assisted reproductive technology. MAIN OUTCOME MEASURE(S): Oocyte survival after thawing. Fertilization, cleavage, and embryo development into blastocyst stage. RESULT(S): Four of six mature (metaphase II) frozen oocytes survived after thawing. Three of them were injected: these three oocytes were fertilized, and one developed into a five-cell embryo on day 3. This embryo developed into blastocyst and was transferred on day 6. A healthy female neonate weighing 3,410 g was born. CONCLUSION(S): After 5 years of storage in liquid nitrogen, cryopreserved oocytes with a slow cooling-rapid thawing protocol can develop in vitro to blastocyst stage and produce a live birth. PMID- 18402941 TI - Glycodelin expression in the endometrium of healthy women and in the eutopic and ectopic endometrium of women with endometriosis. AB - OBJECTIVE: To analyze the expression of the glycodelin gene to better understand the molecular environment of endometriotic lesions and to elucidate the potential mechanisms that underlie the complex physiopathology of endometriosis. DESIGN: Prospective laboratory study. SETTING: University hospital. PATIENT(S): Eleven healthy fertile women and 17 patients with endometriosis in the early proliferative phase of the menstrual cycle. INTERVENTION(S): Endometrial biopsy specimens were obtained from the endometrium of healthy women without endometriosis (controls) and from eutopic and ectopic endometrium tissues (pelvic and ovarian endometriotic implants) of endometriosis patients. MAIN OUTCOME MEASURE(S): The glycodelin relative expression level by real-time polymerase chain reaction (PCR) analysis. RESULT(S): The glycodelin down-regulation found in the endometriotic lesions was 332.26 and 123.17-fold lower, respectively, when compared with the eutopic tissue and the control endometrium. CONCLUSION(S): Glycodelin may be one of the molecules that contributes to the loss of cellular homeostasis in endometriotic lesions. PMID- 18402942 TI - Conservative management of large uterine arteriovenous malformation: case report. AB - Uterine arteriovenous malformation is a rare condition that may be associated with uterine bleeding. Eventually morphologic aspects of the lesions induce unnecessary aggressive surgery; we report here a case of successful expectant management. PMID- 18402943 TI - Dichorionic triamniotic triplet pregnancy with monozygotic twins discordant for trisomy 13 after preimplantation genetic screening: case report. AB - OBJECTIVE: To report the first dichorionic triamniotic triplet pregnancy discordant for trisomy 13 after in vitro fertilization (IVF) treatment with preimplantation genetic screening (PGS). DESIGN: Case report. SETTING: Private IVF center. PATIENT(S): A 40-year-old para 1+6 woman. INTERVENTION(S): IVF combined with PGS for chromosomes 13, 16, 18, 21, and 22, resulting in the transfer of two embryos. MAIN OUTCOME MEASURE(S): Prenatal fetal ultrasonography revealed a dichorionic triamniotic triplet pregnancy. An amniocentesis, performed at 15-weeks' gestation, confirmed that the singleton and one monozygotic twin were normal but the other monozygotic twin was trisomy 13. RESULT(S): After diagnosis and counseling, selective termination of the trisomy 13 monozygotic twin was performed at 16 weeks and 4 days. At 18 weeks and 4 days the co-twin died. A healthy boy was delivered by elective caesarean section at 36-weeks' gestation. CONCLUSION(S): Assisted reproductive techniques that breach the embryo's zona pellucida such as assisted hatching and PGS embryo biopsy increase the incidence of monozygotic twins. Due to high levels of mosaicism in human preimplantation embryos, PGS cannot ensure that embryos diagnosed as normal and selected for transfer do not contain abnormal cells. Hence, further reports of discordant monozygotic twins following PGS are expected, emphasizing the need for appropriate counseling of patients wishing to embark on an IVF/PGS treatment cycle. PMID- 18402945 TI - The value of vascular endothelial growth factor, pregnancy-associated plasma protein-A, and progesterone for early differentiation of ectopic pregnancies, normal intrauterine pregnancies, and spontaneous miscarriages. AB - OBJECTIVE: To evaluate the capacity of vascular endothelial growth factor (VEGF), pregnancy-associated plasma protein-A (PAPP-A), and progesterone (P) to discriminate ectopic pregnancies (EP) from nonectopic ones. DESIGN: Prospective, case-controlled study. SETTING: Tertiary care center. PATIENT(S): Twenty-nine women with EP, 29 women with normal intrauterine pregnancy (nIUP), and 28 women with spontaneous miscarriage, all matched for gestational age. INTERVENTION(S): Serum samples were obtained. MAIN OUTCOME MEASURE(S): Serum concentrations of VEGF, PAPP-A, and P were measured. RESULT(S): Serum VEGF concentrations did not show statistically significant differences among women with EP (median, 55.24 pg/mL; range, 0.20-179.24), spontaneous miscarriages (median, 26.24 pg/mL; range, 0.22-365.24), and nIUP (median, 43.24 pg/mL; range, 0.86-101.24). The median level of P was significantly increased in the nIUP group (20.58 ng/mL; range, 13.9-37.04) compared with the other two groups, but there was no statistically significant difference between the spontaneous miscarriage and EP groups. Like P, PAPP-A values were also significantly higher in the nIUP group than in the other two groups, but the difference between PAPP-A values in the EP and spontaneous abortion groups was statistically insignificant. CONCLUSION(S): VEGF, PAPP-A, and P cannot be used to diagnose EPs, but PAPP-A and P can at least be used to differentiate abnormal pregnancies. PMID- 18402944 TI - A randomized placebo-controlled study on the effects of pioglitazone on cortisol metabolism in polycystic ovary syndrome. AB - OBJECTIVE: To investigate possible effects of insulin-sensitizing treatment on cortisol metabolism in insulin-resistant patients with polycystic ovary syndrome (PCOS). DESIGN: Randomized placebo-controlled study. SETTING: Academic tertiary care medical center. PATIENT(S): Thirty insulin-resistant PCOS patients. INTERVENTION(S): Sixteen weeks of pioglitazone (30 mg/day) or placebo treatment. MAIN OUTCOME MEASURE(S): Twenty-four-hour 20 min integrated blood sampling for measurement of cortisol and 24 h urinary excretion of steroid metabolites. Relative 5alpha-reductase activity was evaluated by allotetrahydrocortisol (alloTHF)/THF and androsterone/etiocholanolone (A/E) ratios. Delta values denoted changes during the treatment period (16 weeks--basal). Pyridostigmine growth hormone (GH) stimulation tests were performed, and testosterone (T), dihydrotestosterone (DHT), DHEA, DHEAS, adiponectin, and insulin-like growth factor I (IGF-I) were measured before and after intervention. RESULT(S): Insulin sensitivity, GH, adiponectin, and IGF-I significantly increased during pioglitazone treatment, whereas alloTHF/THF levels significantly decreased. Delta alloTHF/THF levels inversely correlated with Delta adiponectin levels. Delta A/E ratio inversely correlated with Delta IGF-I and Delta peak GH during GH stimulation tests. No significant changes were measured in T, DHT, DHEA, DHEAS, 24 h mean cortisol, or urinary excretion of steroid metabolites. CONCLUSION(S): Pioglitazone decreased relative 5alpha-reductase activity, whereas no significant changes were measured in cortisol levels or urinary cortisol excretion. PMID- 18402946 TI - Overexpression of fucosyltransferase VII (FUT7) promotes embryo adhesion and implantation. AB - OBJECTIVE: To investigate the effect of increased fucosyltransferase VII (FUT7) on sialyl Lewis X (sLeX) synthesis and their impact on embryo implantation. DESIGN: Cell and animal study. SETTING: Liaoning Provincial Core Lab of Glycobiology and Glycoengineering, Department of Biochemistry and Molecular Biology, Dalian Medical University. ANIMAL(S): Adult female mice of Kunming species. INTERVENTION(S): A pIRES2-EGFP-FUT7 expression plasmid containing full length human FUT7 cDNA was constructed and transiently transfected into RL95-2 or injected into the uteri of early pregnant mice. Control subjects received the mock pIRES2-EGFP or anti-sLeX antibody. MAIN OUTCOME MEASURE(S): The expression of FUT7 and sLeX was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and indirect immunofluorescence. The embryo adhesion rate was assayed in vitro with the cocultured RL95-2 and JAR implantation model. In vivo embryo implantation was investigated using a mouse model. RESULT(S): Expression of FUT7 was significantly increased after transfection of pIRES2-EGFP-FUT7 into RL95-2 cells compared with the parental control and mock vector transfectants, as demonstrated by fluorescent microscopy and RT-PCR. Expression of sLeX was also increased in FUT7 overexpressers, as detected by indirect immunofluorescence. Consistent with the elevated expression of FUT7 and sLeX, embryo adhesion rate and embryo implantation competence were significantly increased (P<.01) in both in vitro and in vivo models. CONCLUSION(S): Overexpression of FUT7 up-regulates sLeX synthesis and enhances sLeX-mediated embryo implantation. PMID- 18402947 TI - Letrozole stimulates the growth of human endometrial explants cultured in three dimensional fibrin matrix. AB - OBJECTIVE: To investigate the effects of an aromatase inhibitor, letrozole, on the growth of human endometrium in a three-dimensional fibrin matrix model of endometriosis. DESIGN: Experimental study of human endometrial biopsies in a three-dimensional fibrin matrix culture system. SETTING: Academic research center. PATIENT(S): Eight normal women with benign gynecologic problems. INTERVENTION(S): Endometrial biopsy samples were washed, cut into small pieces, and placed between two layers of fibrin gel in the presence or absence of letrozole in the culture medium. Tissue changes were assessed by histological and immunohistochemical staining using an inverted microscope, image analysis, and a semiquantitative scoring system. MAIN OUTCOME MEASURE(S): Stromal and epithelial cell outgrowth into the fibrin matrix and angiogenesis comprising endothelial cell invasion of the matrix. RESULT(S): Letrozole (0.1 micromol/L, 1 micromol/L, and 10 micromol/L) exerted a significant growth stimulation effect on endometrial tissue in this model. CONCLUSION(S): In contrast to our expectations, letrozole stimulated growth of normal human endometrium in an in vitro model of endometriosis. Normal endometrium may respond differently than endometriotic lesions to therapeutic agents. Our findings should be kept in mind when considering future research to explore new clinical treatments for endometriosis. PMID- 18402948 TI - Isolated follicle-stimulating hormone (FSH) deficiency without mutation of the FSHbeta gene and successful treatment with human menopausal gonadotropin. AB - OBJECTIVE: To describe the case of isolated follicle-stimulating hormone (FSH) deficiency without mutation of the FSHbeta gene. DESIGN: Case report. SETTING: Division of Endocrinology and Metabolism, Department of Internal Medicine, Faculty of Medicine, Kagawa University, Kagawa, Japan. PATIENT(S): A 22-year-old man referred for infertility, azoospermia, and isolated FSH deficiency. INTERVENTION(S): The patient's FSHbeta gene was sequenced. Pituitary function at baseline and after repeated GnRH administration was evaluated. Testicular biopsy was performed. The patient was treated with human menopausal gonadotropin (hMG). MAIN OUTCOME MEASURE(S): Pathologic examination revealed hypospermatogenesis with isolated FSH deficiency without mutation of the FSHbeta gene. RESULT(S): The FSH levels remained below the normal range despite repeated GnRH stimulation. Hypospermatogenesis was confirmed by testicular biopsy. After 6 months of hMG treatment, spermatogenesis was successfully induced. CONCLUSION(S): We report the case of an infertile male with isolated FSH deficiency without any evidence of mutation in the FSHbeta gene. PMID- 18402949 TI - Narrow-band imaging system with magnifying endoscopy for rectal amyloidosis. PMID- 18402951 TI - Flexible endoscopic Killian-Jamieson diverticulotomy and literature review (with video). PMID- 18402950 TI - Air and fluid leak tests after NOTES procedures: a pilot study in a live porcine model (with videos). AB - BACKGROUND: Transluminal access site closure remains a major challenge in natural orifice transluminal endoscopic surgery (NOTES). OBJECTIVE: Our purpose was to develop in vivo leak tests for evaluation of the integrity of transgastric access closure. SETTINGS: Survival experiments on 12 50-kg pigs. DESIGN AND INTERVENTIONS: After a standardized transgastric approach to the peritoneal cavity and peritoneoscopy, the gastric wall incision was closed with T-bars (Wilson-Cook Medical, Winston-Salem, NC) deployed on both sides of the incision and then cinched together. Gastrotomy closure was assessed with air and fluid leak tests. The animals were observed for 1 week and then underwent endoscopic evaluation and necropsy. MAIN OUTCOME MEASUREMENTS: (1) Leak-proof closure of the gastric wall incision. (2) Gastric incision healing 1 week after the procedure. RESULTS: The mean intraperitoneal pressure increased 10.7 +/- 3.7 mm Hg during gastric insufflation when the air leak test was performed before closure compared with 0.9 +/- 0.8 mm Hg after transmural closure of the transgastric access site with T-bars (P < .001). Fluid leak tests demonstrated no leakage of liquid contrast from the stomach into the peritoneal cavity after closure. Necropsy in 1 week confirmed completeness of the gastric closure in all animals with full thickness healing and no spillage of the gastric contents into the peritoneal cavity. LIMITATIONS: Leak tests were only evaluated on an animal model. CONCLUSIONS: Fluid and air leak tests are simple techniques to evaluate in vivo the adequacy of the transluminal access site closure after NOTES procedures. Leak proof gastric closure resulted in adequate tissue approximation and full thickness healing of the gastric wall incision. PMID- 18402952 TI - Esophageal ulceration from high-intensity focused US catheter ablation for pulmonary vein isolation: a novel complication in the treatment of symptomatic refractory atrial fibrillation. PMID- 18402953 TI - Accidental diagnosis of Trichuris trichiura by colonoscopy. PMID- 18402954 TI - Silastic silicone rubber band migration: an unusual complication resolved by endoscopy. PMID- 18402955 TI - Low frequency of bacteremia after an endoscopic resection for large colorectal tumors in spite of extensive submucosal exposure. AB - BACKGROUND: During an EMR or endoscopic submucosal dissection (ESD) for colorectal tumors, an injection needle catheter is passed through the contaminated endoscopic channel and may directly inoculate bacteria into the blood stream during submucosal injection. In addition, extensively exposed submucosa, especially with an ESD, directly contacts colonic luminal bacteria after the procedure, which may increase the risk of bacteremia. However, the incidence of bacteremia after an EMR or ESD for colorectal tumors has not been reported. OBJECTIVE: To evaluate the frequency of bacteremia associated with an EMR or ESD for colon lesions. DESIGN: A prospective study. PATIENTS: A total of 40 patients who underwent a conventional EMR (n = 30), an EMR after circumferential pre-cutting (n = 3), or ESD (n = 7) for colorectal tumors. INTERVENTIONS AND MAIN OUTCOME MEASUREMENTS: Blood cultures were obtained immediately before, 5 minutes after, and 30 minutes after the procedure. Patients were closely monitored for 24 hours after the procedure to detect the development of infectious complications. RESULTS: Blood cultures at baseline and 5 minutes after the procedure were all negative. However, a blood culture at 30 minutes after the procedure showed a positive result in 1 of 40 patients (2.5%). This patient underwent a conventional EMR, and the isolated microorganism was coagulase-negative Staphylococcus, which might be regarded as a contaminant. None of the 40 patients showed any signs or symptoms associated with infection. LIMITATION: The small sample size. CONCLUSIONS: An EMR, or even an ESD, for colon lesions may be considered a low-risk procedure for infectious complications that does not warrant prophylactic administration of antibiotics. PMID- 18402956 TI - Intraductal US in the assessment of tumor involvement to the orifice of the cystic duct by malignant biliary obstruction. AB - BACKGROUND: Tumor involvement to the orifice of the cystic duct (OCD) is a risk factor for cholecystitis after metallic stent (MS) placement. OBJECTIVE: ERCP and intraductal US (IDUS) were used to assess tumor involvement to the OCD. DESIGN: A prospective consecutive study. SETTING: The University of Tokyo Hospital, Japan. PATIENTS AND INTERVENTIONS: Between January 2003 and May 2007, 34 patients with resectable malignant biliary obstruction underwent an ERCP and an IDUS before preoperative biliary drainage. MAIN OUTCOME MEASUREMENTS: ERCP and IDUS findings were confirmed via a histopathologic analysis of surgical specimens. RESULTS: An ERCP showed that the cystic duct was opacified in 22 of 34 patients (64.7%). Eight patients were diagnosed with OCD tumor involvement. In the remaining 12 patients, tumor involvement was impossible to evaluate via an ERCP. An IDUS identified tumor involvement to the OCD in all 34 patients. Tumor involvement to the OCD was diagnosed in 12 patients, including 4 patients with previously negative results during an ERCP. Tumor involvement was confirmed by histopathology of surgical specimens. The sensitivity and specificity of detection were 66.7% (8/12) and 63.6% (14/22), respectively, via ERCP, and 100% (12/12) and 100% (22/22), respectively, via IDUS. LIMITATION: The small sample size. CONCLUSIONS: IDUS, in addition to ERCP, provides an accurate evaluation of tumor involvement to the OCD in patients with malignant biliary obstruction. PMID- 18402957 TI - Successful closure of a rectovaginal fistula by using an endoscopically placed Resolution clip. AB - BACKGROUND: Rectovaginal fistula (RVF) is an abnormal epithelium-lined communication between the wall of the rectum and the posterior vaginal wall. The incidence of RVFs is low and accounts for about 5% of all anorectal fistulas. Women who suffer from an RVF complain of uncontrollable passage of gas or feces from the vagina. This remains a major contributor to morbidity associated with this condition in terms of social, psychologic, and sexual dysfunction. OBJECTIVE: RVFs may be managed both medically and surgically, with the latter being the preferred option. A number of different surgical techniques that pertain to fistula closure were described in various literature; however, very little has been said of much-less-invasive techniques and alternatives to surgical correction if the patient is a poor candidate or prefers not to have surgery. The purpose of our article is to show our approach in treating an RVF, given the fact that our patient was a poor surgical candidate and, moreover, refused more-invasive techniques for treatment. DESIGN: A case report. The patient described in this article is a 77-year-old woman with comorbidities that limited her as a candidate for less-invasive techniques rather than surgery. SETTING: An inpatient at New York Hospital Queens, Flushing, New York. PATIENT: A 77-year-old woman, with a medical history of 2 myocardial infarctions, congestive heart failure, 2 cardiac stents, multiple urinary-tract infections, and diverticulitis, presented to the hospital with a fever of 38.3 degrees C (101 degrees F) for 2 days. On the second day of admission, the patient complained of passing stool and flatus from the vagina. A subsequent workup and sigmoidoscopy revealed an RVF. INTERVENTION: A sigmoidoscopy was performed, and fistula closure was achieved in 2 phases with the use of a Resolution clip. MAIN OUTCOME MEASUREMENTS: Complications and resolution of symptoms after the procedure were the primary end points. RESULTS: No complications were noted post procedure, and the patient has remained free of any complaints now for 12 months. LIMITATIONS: The main limitation of the study is that this is a case report limited to a single patient, and outcomes of the procedure were concluded based on this particular patient. The endoscopic technique described here may be limited to readily visualized fistulas at endoscopy that are amenable to this treatment option. CONCLUSIONS: The technique described, closing an RVF with the use of an endoscopically placed Resolution clip, shows great promise and could be applied to treating uncomplicated fistulas. PMID- 18402958 TI - The new PillCam Colon capsule: difficult colonoscopy? No longer a problem? PMID- 18402959 TI - Affinities of the Swartkrans early Homo mandibles. AB - The southern African early Homo assemblage continues to make important contributions to understanding the systematics, adaptations and evolutionary history of the human genus. However, the taxonomy of this sample is in a state of flux. This study examines the size and shape of the mandibular bodies of Swartkrans SK 15 and SK 45 comparing them with variation in two early Homo taxa (H. habilis sensu lato and H. sapiens erectus). The research aims to clarify their phenetic affinities and systematics through univariate statistics, inferential testing and multivariate analysis employing size (Log-transformed) and shape (Mosimann variables). Neither of them strongly resembles H. habilis sensu lato or H. sapiens erectus, rather, they probably sample a novel species of Homo not seen in East Africa. Moreover, there is considerable morphological variability within the Swartkrans sample and the possibility of more than one novel species being sampled at this site cannot be excluded. PMID- 18402961 TI - Ambient temperature effects on photo induced gonadal cycles and hormonal secretion patterns in Great Tits from three different breeding latitudes. AB - The present study determines how populations of Great Tits (Parus major) breeding in southern, mid and northern European latitudes have adjusted their reproductive endocrinology to differences in the ambient temperature during the gonadal cycle. A study based on long-term breeding data, using the Colwell predictability model, showed that the start of the breeding season has a high predictability ( approximately 0.8-0.9) at all latitudes, and that the environmental information factor (I(e)) progressively decreased from mid Italy (I(e)>4) to northern Finland (I(e)<1). The results indicate that integration of supplementary information, such as ambient temperature, with photoperiodic initial predictive information (day length), becomes progressively more important in maintaining the predictability of the breeding season with decreasing latitude. This hypothesis was verified by exposing photosensitive Great Tits from northern Norway, southern Sweden and northern Italy to sub-maximal photo-stimulatory day lengths (13L:11D) under two different ambient temperature regimes (+4 degrees C and +20 degrees C). Changes in testicular size, plasma levels of LH and testosterone were measured. The main results were: (1) Initial testicular growth rate, as well as LH secretion, was affected by temperature in the Italian, but not in birds from the two Scandinavian populations. (2) Maximum testicular size, maximum LH and testosterone levels were maintained for a progressively shorter period of time with increasing latitude, regardless of whether the birds were kept on a low or a high ambient temperature. (3) In birds from all latitudes, the development of photorefractoriness, as indicated by testicular regression and a decrease in plasma levels of LH and testosterone, started much earlier (with the exception for LH Great Tits from northern Scandinavia) when kept on +20 degrees C than when kept on +4 degrees C. The prolonging effects of a low temperature was more pronounced in Mediterranean birds, than in birds from Scandinavia, and more pronounced in Great Tits from southern Scandinavia than in Great Tits from northern Scandinavia. Ecological implications of the results are discussed, as well as possible impact of global warming on the breeding success of European Great Tits from different breeding latitudes. PMID- 18402962 TI - System identification of two-dimensional continuous-time systems using wavelets as modulating functions. AB - In this paper, parameter identification of two-dimensional continuous-time systems via two-dimensional modulating functions is proposed. In the proposed method, trigonometric functions and sine-cosine wavelets are used as modulating functions. By this, a partial differential equation on the finite-time intervals is converted into an algebraic equation linear in parameters. The parameters of the system can then be estimated using the least square algorithms. The underlying computations utilize a two-dimensional fast Fourier transform algorithm, without the need for estimating the unknown initial or boundary conditions, at the beginning of each finite-time interval. Numerical simulations are presented to show the effectiveness of the proposed algorithm. PMID- 18402960 TI - Classical androgen receptors in non-classical sites in the brain. AB - Androgen receptors are expressed in many different neuronal populations in the central nervous system where they often act as transcription factors in the cell nucleus. However, recent studies have detected androgen receptor immunoreactivity in neuronal and glial processes of the adult rat neocortex, hippocampal formation, and amygdala as well as in the telencephalon of eastern fence and green anole lizards. This review discusses previously published findings on extranuclear androgen receptors, as well as new experimental results that begin to establish a possible functional role for androgen receptors in axons within cortical regions. Electron microscopic studies have revealed that androgen receptor immunoreactive processes in the rat brain correspond to axons, dendrites and glial processes. New results show that lesions of the dorsal CA1 region by local administration of ibotenic acid reduce the density of androgen receptor immunoreactive axons in the cerebral cortex and the amygdala, suggesting that these axons may originate in the hippocampus. Androgen receptor immunoreactivity in axons is also decreased by the intracerebroventricular administration of colchicine, suggesting that androgen receptor protein is transported from the perikaryon to the axons by fast axonal transport. Androgen receptors in axons located in the cerebral cortex and amygdala and originating in the hippocampus may play an important role in the rapid behavioral effects of androgens. PMID- 18402963 TI - Round versus flat: bone cell morphology, elasticity, and mechanosensing. AB - There is increasing evidence that cell function and mechanical properties are closely related to morphology. However, most in vitro studies investigate flat adherent cells, which might not reflect physiological geometries in vivo. Osteocytes, the mechanosensors in bone, reside within ellipsoid containment, while osteoblasts adhere to flatter bone surfaces. It is unknown whether morphology difference, dictated by the geometry of attachment is important for cell rheology and mechanosensing. We developed a novel methodology for investigating the rheology and mechanosensitivity of bone cells under different morphologies using atomic force microscopy and our two-particle assay for optical tweezers. We found that the elastic constant of MLO-Y4 osteocytes when flat and adherent (>1 kPa) largely differed when round but partially adherent (<1 kPa). The elastic constant of round suspended MLO-Y4 osteocytes, MC3T3-E1 osteoblasts, and primary osteoblasts were similarly <1 kPa. The mechanosensitivity of round suspended MLO-Y4 osteocytes was investigated by monitoring nitric oxide (NO) release, an essential signaling molecule in bone. A preliminary observation of high NO release from round suspended MLO-Y4 osteocytes in response to approximately 5 pN force is reported here, in contrast with previous studies where flat cells routinely release lesser NO while being stimulated with higher force. Our results suggest that a round cellular morphology supports a less stiff cytoskeleton configuration compared with flat cellular morphology. This implies that osteocytes take advantage of their ellipsoid morphology in vivo to sense small strains benefiting bone health. Our assay provides novel opportunities for in vitro studies under a controlled suspended morphology versus commonly studied adherent morphologies. PMID- 18402964 TI - On the cytoskeleton and soft glassy rheology. AB - The cytoskeleton is a complex structure within the cellular corpus that is responsible for the main structural properties and motilities of cells. A wide range of models have been utilized to understand cytoskeletal rheology and mechanics (see e.g. [Mofrad, M., Kamm, R., 2006. Cytoskeletal Mechanics: Models and Measurements. Cambridge University Press, Cambridge]). From this large collection of proposed models, the soft glassy rheological model (originally developed for inert soft glassy materials) has gained a certain traction in the literature due to the close resemblance of its predictions to certain mechanical data measured on cell cultures [Fabry, B., Maksym, G., Butler, J., Glogauer, M., Navajas, D., Fredberg, J., 2001. Scaling the microrheology of living cells. Physical Review Letters 87, 14102]. We first review classical linear rheological theory in a concise fashion followed by an examination of the soft glassy rheological theory. With this background we discuss the observed behavior of the cytoskeleton and the inherent limitations of classical rheological models for the cytoskeleton. This then leads into a discussion of the advantages and disadvantages presented to us by the soft glassy rheological model. We close with some comments of caution and recommendations on future avenues of exploration. PMID- 18402965 TI - Optimisation of high bar circling technique for consistent performance of a triple piked somersault dismount. AB - The dismount from the high bar is one of the most spectacular skills performed in Men's Artistic Gymnastics. Hiley and Yeadon [2005. Maximal dismounts from high bar. Journal of Biomechanics 38, 2221-2227] optimised the technique in the backward giant circle prior to release using a computer simulation model to show that a gymnast could generate sufficient linear and angular momentum to perform a triple piked backward somersault dismount with a sufficiently large release window (the period of time during which the gymnast could release the bar and successfully complete the dismount). In the present study, it was found that when the timing of the actions at the hip and shoulder joints from the optimum simulation were perturbed by 30ms the resulting simulation could no longer meet the criteria for sufficient aerial rotation and release window. Since it is to be expected that a gymnast's technique can cope with small errors in timing for consistent performance, a requirement of robustness to timing perturbations should be included within the optimisation process. When the technique in the backward giant circle was optimised to be robust to 30ms perturbations, it was found that sufficient linear and angular momentum for a triple piked dismount could be achieved with a realistic release window. PMID- 18402966 TI - Defining and evaluating wrapping surfaces for MRI-derived spinal muscle paths. AB - Muscle paths can be approximated in biomechanical models by wrapping the path around geometric objects; however, the process for selecting and evaluating wrapping surface parameters is not well defined, especially for spinal muscles. In this study, we defined objective methods to select the shape, orientation, size and location of wrapping surfaces and evaluated the wrapping surfaces using an error metric based on the distance between the modeled muscle path and the centroid path from magnetic resonance imaging (MRI). We applied these methods and the error metric to a model of the neck musculature, where our specific goals were (1) to optimize the vertebral level at which to place a single wrapping surface per muscle; and (2) to define wrapping surface parameters in the neutral posture and evaluate them in other postures. Detailed results are provided for the sternocleidomastoid and the semispinalis capitis muscles. For the sternocleidomastoid, the level where the wrapping surface was placed did not significantly affect the error between the modeled path and the centroid path; use of wrapping surfaces defined from the neutral posture improved the representation of the muscle path compared to a straight line in all postures except contralateral rotation. For the semispinalis capitis, wrapping surfaces placed at C3 or C4 resulted in lower error compared to other levels; and the use of wrapping surfaces significantly improved the muscle path representation in all postures. These methods will be used to improve the estimates of muscle length, moment arm and moment-generating capacity in biomechanical models. PMID- 18402967 TI - Enhancing sensitivity of liquid chromatographic/ion-trap tandem mass spectrometric determination of estrogens by on-line pre-column derivatization. AB - A rapid and sensitive liquid chromatographic/ion-trap mass spectrometric method has been developed for the simultaneous determination of estriol, alpha-, beta estradiol, 17alpha-ethynylestradiol and estrone. The substances were extracted from river water samples (typically 250 ml) and cleaned-up using two-stage solid phase extraction (SPE). 12-(Difluoro-1,3,5-triazinyl)-benz[f]isoindolo [1,2b][1,3]benzothiazolidine was applied as a derivatizing agent. The recovery for each compounds ranged from 75 to 88% and the repeatability represented as RSDs ranged from 5.6 to 8.4%. Limits of detection (LOD, 3 x S/N) were 67-285 pg/l. PMID- 18402968 TI - Electrocatalytic oxidation of ethylene glycol on Pt and Pt-Ru nanoparticles modified multi-walled carbon nanotubes. AB - The synthesis and characterization of catalysts based on nanomaterials, supported on multi-walled carbon nanotubes (CNT) for ethylene glycol (EG) oxidation is investigated. Platinum (Pt) and platinum-ruthenium (Pt-Ru) nanoparticles are deposited on surface-oxidized multi-walled carbon nanotubes [Pt/CNT; Pt-Ru/CNT] by the aqueous solution reduction of the corresponding metal salts with glycerol. The electrocatalytic properties of the modified electrodes for oxidation of ethylene glycol in acidic solution have been studied by cyclic voltammetry (CV), and excellent activity is observed. This may be attributed to the small particle size of the metal nanoparticles, the efficacy of carbon nanotubes acting as good catalyst support and uniform dispersion of nanoparticles on CNT surfaces. The nature of the resulting nanoparticles decorated multiwalled carbon nanotubes are characterized by scanning electron microscopy (SEM) and transmission electron microscopic (TEM) analysis. The cyclic voltammetry response indicates that Pt Ru/CNT catalyst displays a higher performance than Pt/CNT, which may be due to the efficiency of the nature of Ru species in Pt-Ru systems. The fabricated Pt and Pt-Ru nanoparticles decorated CNT electrodes shows better catalytic performance towards ethylene glycol oxidation than the corresponding nanoparticles decorated carbon electrodes, demonstrating that it is more promising for use in fuel cells. PMID- 18402969 TI - Surface modification of polyaniline using tetraethyl orthosilicate. AB - A well-dispersible conducting polyaniline/silica hybrid is prepared by the hydrolysis and condensation of tetraethyl orthosilicate (TEOS) on the surface of polyaniline in water/ethanol solution. It provides a simple and environmentally sound route for preparing the processable conducting polyaniline/silica hybrid at the nanometre level. The conductivity of polyaniline/silica hybrid is 2.43 S cm( 1) at 25 degrees C, and its powder is easily dispersed in the anhydrous ethanol or aqueous solution without any stabilizer. In addition, the structure, morphology and cyclic voltammorgram of this hybrid are also reported. PMID- 18402970 TI - Influence of organic matter removal on competitive and noncompetitive adsorption of copper and zinc in acid soils. AB - We studied competitive and noncompetitive adsorption of copper and zinc in four acid soils, and compared the behavior of the two metals in untreated samples and samples treated with hydrogen peroxide to remove organic matter in the soil. Copper exhibited stronger competitive adsorption than zinc in the untreated samples. However, removal of organic matter reduced copper adsorption to a greater extent than zinc adsorption, the two metals exhibiting a more similar adsorption pattern than the untreated samples. The presence of copper dramatically reduced zinc competitive adsorption in untreated samples; on the other hand, that of zinc only resulted in slightly reduced competitive adsorption of copper. The hydrogen peroxide treatment decreased competitive adsorption in both metals; however, copper continued to be more efficient than zinc in competing for binding sites on adsorbing surfaces. Desorption of Cu occurred much less readily than desorption of Zn and hysteresis is apparent especially for Cu. PMID- 18402971 TI - Preparation of PLLA/PLGA microparticles using solution enhanced dispersion by supercritical fluids (SEDS). AB - In this work, poly(L-lactic acid)/poly(lactide-co-glycolide) (PLLA/PLGA) microparticles were prepared using the technique of solution-enhanced dispersion by supercritical fluids (SEDS). For comparison, separate PLLA and PLGA microparticles were also produced by the same SEDS process. The produced microparticles were characterized by scanning electron microscopy, laser particle size analyzer, X-ray diffraction, differential scanning calorimetry, Fourier transform infrared spectroscopy, and gas chromatography. Results indicate that PLLA/PLGA microparticles possess sphere-like shapes with smooth surfaces. The mean particle size of PLLA/PLGA microparticles ranges from 1.76 to 2.15 microm, depending on the feeding ratio of PLLA to PLGA used in the SEDS process. The crystallinity of PLLA/PLGA microparticles decreases after the SEDS processing, so that the produced microparticles are in an amorphous state. Pure PLGA was hard to precipitate in small, fine microparticle form without the presence of PLLA. A model drug, paclitaxel, was encapsulated into PLLA/PLGA microparticles by the same SEDS process, and the in vitro release rate of paclitaxel from these PLLA/PLGA composites could be modulated by variation of the mixing ratio PLLA:PLGA. The prepared microparticles have negligible residual organic solvent. Drug-loaded PLLA/PLGA microparticles produced by SEDS have potential as an advanced colloidal suspension for pharmaceutical applications. PMID- 18402972 TI - Transport of poly-L-lysine into oppositely charged poly(acrylic acid) microgels and its effect on gel deswelling. AB - The interaction between poly-L-lysine (pLys) and oppositely charged poly(acrylic acid) (pAA) microgels (O approximately 80-120 microm) was studied by micromanipulator-assisted light microscopy and confocal laser scanning microscopy. The aim of this study was to investigate effects of peptide size, pH, and salt concentration on binding, transport, and distribution of pLys in pAA microgel particles and thereby also to outline the details of the gel deswelling in response to pLys binding and incorporation. Both peptide distribution and gel deswelling kinetics were found to be strongly influenced by the pLys molecular weight, originating partly from limited entry of large peptides into the gel particle core. Also pH was shown to influence both deswelling and pLys incorporation kinetics, with a decreased deswelling rate observed with increasing pH. These effects are determined by a complex interplay between the pH-dependence of both pLys and the gel network, also influencing volume transitions of the latter. Finally, salt concentration was shown to have a significant effect on both gel deswelling rate and pLys transport, with an increased electrolyte concentration resulting in decreased deswelling rate but also in an increased peptide transport rate within the microgel particles. PMID- 18402973 TI - Fabrication of silica nanotubes using silica coated multi-walled carbon nanotubes as the template. AB - Silica nanotubes were synthesized using the multi-walled carbon nanotubes (MWCNTs) as the template material. First, we prepared silica coated MWCNT composites by surface oxidation of MWCNTs using KMnO(4) in the presence of a phase transfer catalyst and followed by grafting of 2-aminoethyl 3-aminopropyl trimethoxy silane, AEAPS. The amine groups in grafted AEAPS on MWCNTs could activate the silica shell formation by acid-base interaction. The synthesized silica was formed a uniform layer on MWCNTs with a controllable thickness and possessed sturdy 3-dimensional stability. After calcinations at 800 degrees C, the inner MWCNTs of the composite were completely decomposed and the outer silica shell layer maintained without distortion of its original shape. Finally, we could obtain the silica nanotubes having 13.0 nm of average layer thickness. PMID- 18402974 TI - Extruded vesicles of dioctadecyldimethylammonium bromide and chloride investigated by light scattering and cryogenic transmission electron microscopy. AB - Combined dynamic and static light scattering (DLS, SLS) and cryogenic transmission electron microscopy (cryo-TEM) were used to investigate extruded cationic vesicles of dioctadecyldimethylammonium chloride and bromide (DODAX, X being Cl(-) or Br(-)). In salt-free dispersions the mean hydrodynamic diameter, D(h), and the weight average molecular weight, M(w), are larger for DODAB than for DODAC vesicles, and both D(h) and M(w) increase with the diameter (varphi) of the extrusion filter. NaCl (NaBr) decreases (increases) the DODAB (DODAC) vesicle size, reflecting the general trend of DODAB to assemble as larger vesicles than DODAC. The polydispersity index is lower than 0.25, indicating the dispersions are rather polydisperse. Cryo-TEM micrographs show that the smaller vesicles are spherical while the larger ones are oblong or faceted, and the vesicle samples are fairly polydisperse in size and morphology. They also indicate that the vesicle size increases with phi and DODAB assembles as larger vesicles than DODAC. Lens-shaped vesicles were observed in the extruded preparations. Both light scattering and cryo-TEM indicate that the vesicle size is larger or smaller than phi when phi is smaller or larger than the optimal phi approximately 200 nm. PMID- 18402975 TI - Strategies to control the particle size distribution of poly-epsilon-caprolactone nanoparticles for pharmaceutical applications. AB - In this work turbulent precipitation through solvent displacement for the production of poly-epsilon-caprolactone (PCL) nanoparticles is investigated; two different PCL molecular weights have been employed, using acetone and water as solvent and anti-solvent, respectively. The main important thermodynamic and kinetic parameters, such as solubility and interfacial tension of PCL in water acetone mixtures, are determined and the effect of the process operating conditions on the final particle size distribution is also investigated. Particles produced under different conditions into a Confined Impinging Jets Reactor (CIJR) were characterized by Dynamic Light Scattering, Zeta potential measurements and Scanning Electronic Microscopy. Results clearly show the strong effect of mixing on the particle size distribution and how mixing must be controlled in order to obtain a product with particular characteristics. Eventually the measured thermodynamic and kinetic parameters are used to interpret the obtained experimental data. PMID- 18402976 TI - High-reproducible flow cytometric endothelial progenitor cell determination in human peripheral blood as CD34+/CD144+/CD3- lymphocyte sub-population. AB - Although determination of circulating endothelial progenitor cell (EPC) in peripheral blood by flow cytometry is an emerging marker for cardiovascular medicine, a common standardized protocol is still not available, due to the low numbers achieved in peripheral blood. In the present paper we describe a novel technique for EPC quantification as CD34+/CD144+/CD3- cells within the lymphocyte gate, which increases the percentages of EPC positivity described before and also offers high intra-assay reproducibility. These improvements are based on a gating strategy for big-sized lymphocytes, smooth fixation and cytometric clearance of CD3+ lymphocytes (T-cells). This last procedure is able to increase intra-assay Pearson's correlation from 0.8517 to 0.8908. Therefore, the technical setting described here offers a high-performance and clinically oriented EPC determination strategy in human peripheral blood. PMID- 18402977 TI - Quantitative determination of humanized monoclonal antibody rhuMAb2H7 in cynomolgus monkey serum using a Generic Immunoglobulin Pharmacokinetic (GRIP) assay. AB - Preclinical pharmacokinetic (PK) assays are important to help evaluate the safety and efficacy of a potential biotherapeutic before clinical studies. The assay typically requires a biotherapeutic-specific reagent to minimize matrix effects especially when the host species are non-human primates such as cynomolgus monkeys and the biotherapeutic is a humanized monoclonal antibody (MAb). Recombinant humanized mAb 2H7 (rhuMAb2H7) binds to the extracellular domain of CD20 that is expressed on B cells and results in B cell depletion. It is currently being evaluated for its therapeutic potential in rheumatoid arthritis (RA) in clinical studies. During the early development of rhuMAb2H7, a cynomolgus monkey PK assay was needed to help assess the pharmacokinetic parameters of rhuMAb2H7 in a pilot cynomolgus monkey study. However, development of a cynomolgus monkey PK assay was challenging due to lack of rhuMAb2H7-specific reagents. Here we describe an alternative method for detection of rhuMAb2H7 in cynomolgus monkey serum using polyclonal antibodies against human IgGs. This assay quantifies rhuMAb2H7 in 10% cynomolgus monkey serum with high sensitivity, accuracy, and precision. This assay successfully supported the rhuMAb2H7 development, and has the potential to be used to quantify other humanized MAb biotherapeutics in serum from a variety of non-human species. PMID- 18402978 TI - A large collapsed-state RNA can exhibit simple exponential single-molecule dynamics. AB - The process of large RNA folding is believed to proceed from many collapsed structures to a unique functional structure requiring precise organization of nucleotides. The diversity of possible structures and stabilities of large RNAs could result in non-exponential folding kinetics (e.g. stretched exponential) under conditions where the molecules have not achieved their native state. We describe a single-molecule fluorescence resonance energy transfer (FRET) study of the collapsed-state region of the free energy landscape of the catalytic domain of RNase P RNA from Bacillus stearothermophilus (C(thermo)). Ensemble measurements have shown that this 260 residue RNA folds cooperatively to its native state at >or=1 mM Mg(2+), but little is known about the conformational dynamics at lower ionic strength. Our measurements of equilibrium conformational fluctuations reveal simple exponential kinetics that reflect a small number of discrete states instead of the expected inhomogeneous dynamics. The distribution of discrete dwell times, collected from an "ensemble" of 300 single molecules at each of a series of Mg(2+) concentrations, fit well to a double exponential, which indicates that the RNA conformational changes can be described as a four state system. This finding is somewhat unexpected under [Mg(2+)] conditions in which this RNA does not achieve its native state. Observation of discrete well defined conformations in this large RNA that are stable on the seconds timescale at low [Mg(2+)] (<0.1 mM) suggests that even at low ionic strength, with a tremendous number of possible (weak) interactions, a few critical interactions may produce deep energy wells that allow for rapid averaging of motions within each well, and yield kinetics that are relatively simple. PMID- 18402979 TI - Modulation by substrates of the interaction between the HasR outer membrane receptor and its specific TonB-like protein, HasB. AB - TonB is a cytoplasmic membrane protein required for active transport of various essential substrates such as heme and iron siderophores through the outer membrane receptors of Gram-negative bacteria. This protein spans the periplasm, contacts outer membrane transporters by its C-terminal domain, and transduces energy from the protonmotive force to the transporters. The TonB box, a relatively conserved sequence localized on the periplasmic side of the transporters, has been shown to directly contact TonB. While Serratia marcescens TonB functions with various transporters, HasB, a TonB-like protein, is dedicated to the HasR transporter. HasR acquires heme either freely or via an extracellular heme carrier, the hemophore HasA, that binds to HasR and delivers heme to the transporter. Here, we study the interaction of HasR with a HasB C-terminal domain and compare it with that obtained with a TonB C-terminal fragment. Analysis of the thermodynamic parameters reveals that the interaction mode of HasR with HasB differs from that with TonB, the difference explaining the functional specificity of HasB for HasR. We also demonstrate that the presence of the substrate on the extracellular face of the transporter modifies, via enthalpy-entropy compensation, the interaction with HasB on the periplasmic face. The transmitted signal depends on the nature of the substrate. While the presence of heme on the transporter modifies only slightly the nature of interactions involved between HasR and HasB, hemophore binding on the transporter dramatically changes the interactions and seems to locally stabilize some structural motifs. In both cases, the HasR TonB box is the target for those modifications. PMID- 18402981 TI - A model for influenza with vaccination and antiviral treatment. AB - Compartmental models for influenza that include control by vaccination and antiviral treatment are formulated. Analytic expressions for the basic reproduction number, control reproduction number and the final size of the epidemic are derived for this general class of disease transmission models. Sensitivity and uncertainty analyses of the dependence of the control reproduction number on the parameters of the model give a comparison of the various intervention strategies. Numerical computations of the deterministic models are compared with those of recent stochastic simulation influenza models. Predictions of the deterministic compartmental models are in general agreement with those of the stochastic simulation models. PMID- 18402980 TI - Integrating cell-cycle progression, drug penetration and energy metabolism to identify improved cancer therapeutic strategies. AB - The effectiveness of chemotherapeutic drugs in tumors is reduced by multiple effects including drug diffusion and variable susceptibility of local cell populations. We hypothesized that quantifying the interactions between drugs and tumor microenvironments could be used to identify more effective anti-cancer strategies. To test this hypothesis we created a mathematical model that integrated intracellular metabolism, nutrient and drug diffusion, cell-cycle progression, cellular drug effects, and drug pharmacokinetics. To our knowledge, this is the first model that combines these elements and has coupled them to experimentally derived parameters. Drug cytotoxicity was assumed to be cell-cycle phase specific, and progression through the cell cycle was assumed to be dependent on ATP generation. The model consisted of a coupled set of nonlinear partial differential, ordinary differential and algebraic equations with an outer free boundary, which was solved using orthogonal collocation on a moving grid of finite elements. Model simulations showed the existence of an optimum drug diffusion coefficient: a low diffusivity prevents effective penetration before the drug is cleared from the blood and a high diffusivity limits drug retention. This result suggests that increasing the molecular weight of the anti-cancer drug paclitaxel from 854 to approximately 20,000 by nanoparticle conjugation would improve its efficacy. The simulations also showed that fast growing tumors are less responsive to therapy than are slower tumors with more quiescent cells, demonstrating the competing effects of regrowth and cytotoxicity. The therapeutic implications of the simulation results are that (1) monolayer cultures are inadequate for accurately determining therapeutic effects in vitro, (2) decreasing the diffusivity of paclitaxel could increase its efficacy, and (3) measuring the proliferation fraction in tumors could enhance the prediction of therapeutic efficacy. PMID- 18402982 TI - Association between risk of myopathy and cholesterol-lowering effect: a comparison of all statins. AB - In the present study, we examined the mechanisms underlying the cytotoxicity of pitavastatin, a new statin, and we compared the in vitro potencies of muscle cytotoxicity using a prototypic embryonal rhabdomyosarcoma cell line (RD cells), a typical side effect of statins and compared the cholesterol-lowering effects of statins using Hep G2 hepatoma cells. Pitavastatin reduced the number of viable cells and caused caspase-9 and -3/7 activation in a time- and concentration dependent manner. The comparison of cytotoxities of statins showed that statins significantly reduced cell viability and markedly enhanced activity of caspase 3/7 in concentration-dependent manner. On the other hand, the effects of hydrophilic statins, pravastatin, rosuvastatin were very weak. The rank order of cytotoxicity was cerivastatin > simvastatin acid> fluvastatin > atorvastatin > lovastatin acid > pitavastatin >> rosuvastatin, pravastatin. Statin-induced cytotoxicity is associated with these partition coefficients. On the other hand, the cholesterol-lowering effect of statins did not correlate with these partition coefficients and cytotoxicity. Thus, it is necessary to consider the association between risk of myopathy and cholesterol-lowering effect of a statin for precise use of statins. PMID- 18402983 TI - Chronic mild stress impairs cognition in mice: from brain homeostasis to behavior. AB - Exposure to chronic stress in rodents and psychosocial stress in humans has been shown to alter cognitive functions and has been linked to the pathophysiology of mood disorders. The purpose of the present study was to investigate effects and possible mechanisms of a chronic mild stress (CMS) procedure on cognitive behaviors in Swiss albino mice using the object recognition test (ORT) and object location test (OLT). Results showed that CMS exposure impaired cognitive performance and produced amnesia of acquired information in both ORT and OLT. Furthermore, the cognitive impairment was coexistent with increased plasma levels of interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha), as well as with enhanced plasma levels of corticosterone (CORT), corticotrophin-releasing hormone (CRH) and adrenocorticotrophic hormone (ACTH). In addition, severe neuronal cell damage was found, as bromodeoxyuridine (BrdU) positive cells and the expression of brain derived neurotrophic factor (BDNF) in dentate gyrus (DG) of hippocampus were decreased after 5 weeks CMS procedure. Taken together, these findings indicated that CMS exposure-induced impairment of cognitive behaviors might be attributed to the stress-related alterations in brain homeostasis that were reflected in changes in the neuroimmune and neuroendocrine systems as well as in neurogenesis. PMID- 18402984 TI - Benthic denitrification and organic matter mineralization in intertidal flats of an enclosed coastal inlet, Ago Bay, Japan. AB - Denitrification (as N(2) flux) and organic matter mineralization (as O(2) uptake) were simultaneously measured in the same set of core sediments from a natural sandy and a constructed muddy tidal flat of Ago Bay, Japan. Denitrification rates at both tidal flats fluctuated between ca. 2-20 micromol N(2)m(-2)h(-1) without showing a clear seasonal pattern, and appeared to be substrate limited as NO(3)( ) enrichment (final concentration ca. 225 microM) caused prompt and similar enhancements of ca. 10-folds. Organic matter mineralization rates were markedly higher at constructed muddy flat compared to those of natural sandy flat, especially in summer, and exhibited pronounced temperature dependent (p<0.01) seasonality for both tidal flats. O(2) uptake rates were generally ca. 2-3 order greater than respective denitrification rates indicating dominance of mineralization processes over N(2) losses. PMID- 18402985 TI - Endogenous neuroprotection: mitochondria as gateways to cerebral preconditioning? AB - From single to multicellular organisms, protective mechanisms have evolved against endogenous and exogenous noxious stimuli. Preconditioning paradigms, in which stimulation below the threshold of injury results in subsequent protection of the brain, have played an important role in elucidating such endogenous protective mechanisms. Consequently, over the past decades numerous signaling pathways have been discovered by which the brain senses and reacts to such insults as neurotoxins, substrate deprivation, or inflammation. Research on preconditioning is aimed at understanding endogenous neuroprotection to boost it, or to supplement its effectors therapeutically once damage to the brain has occurred, such as after stroke or brain trauma. Another goal of establishing preconditioning protocols is to induce endogenous neuroprotection in anticipation of incipient brain damage. Currently several endogenous neuroprotectants are being investigated in controlled clinical trials. In the present review we will give a short overview on the signals, sensors, transducers, and effectors of endogenous neuroprotection. We will first focus on common mechanisms, on which pathways of endogenous neuroprotection converge, and in particular on mitochondria, which may be considered master integrators of endogenous neuroprotection. We will then discuss various applications of preconditioning, including pharmacological and anesthetic preconditioning, as well as postconditioning, and explore the prospects of endogenous neuroprotective therapeutic approaches. PMID- 18402987 TI - N-methyl-D-aspartate mechanisms in depolarization of augmenting expiratory neurons during the expulsive phase of fictive cough in decerebrate cats. AB - Cough reflex is characterized by a large expulsive phase for expelling the mucus or particles from the airway. The present study investigated the involvement of N methyl-D-aspartate (NMDA) mechanisms in the expulsive phase of cough reflex using decerebrate and paralyzed cats. A fictive cough was induced by repetitive stimulation of the superior laryngeal nerve, which was characterized by an increased inspiratory discharge in the phrenic nerve (the stage 1 of fictive cough; SC1) and large spindle-shaped discharge in the iliohypogastric nerve (the stage 2 of fictive cough; SC2). Intravenous injection of an antagonist of NMDA receptors, dizocilpine (0.1mg/kg), increased the threshold intensity of stimulation for inducing a fictive cough. The SC2 iliohypogastric response was more vulnerable to dizocilpine than the SC1 phrenic response. Membrane potential of augmenting expiratory (aug-E) neurons was recorded from the caudal ventral respiratory group. Aug-E neurons showed a large depolarization with a high frequency discharge during the SC2 in major cases (n=35) and hyperpolarization in minor cases (n=6). Dizocilpine inhibited the occurrence of these SC2 responses of aug-E neurons without any effect on the basal respiratory fluctuations of membrane potential. This drug had no significant effect on waves of excitatory and inhibitory postsynaptic potentials evoked in aug-E neurons by single pulse stimulation of the SLN. The present results demonstrated that NMDA mechanisms contribute preferentially to the expulsive phase response in aug-E neurons during fictive cough reflex. PMID- 18402988 TI - Seeing direct and averted gaze activates the approach-avoidance motivational brain systems. AB - Gaze direction is known to be an important factor in regulating social interaction. Recent evidence suggests that direct and averted gaze can signal the sender's motivational tendencies of approach and avoidance, respectively. We aimed at determining whether seeing another person's direct vs. averted gaze has an influence on the observer's neural approach-avoidance responses. We also examined whether it would make a difference if the participants were looking at the face of a real person or a picture. Measurements of hemispheric asymmetry in the frontal electroencephalographic activity indicated that another person's direct gaze elicited a relative left-sided frontal EEG activation (indicative of a tendency to approach), whereas averted gaze activated right-sided asymmetry (indicative of avoidance). Skin conductance responses were larger to faces than to control objects and to direct relative to averted gaze, indicating that faces, in general, and faces with direct gaze, in particular, elicited more intense autonomic activation and strength of the motivational tendencies than did control stimuli. Gaze direction also influenced subjective ratings of emotional arousal and valence. However, all these effects were observed only when participants were facing a real person, not when looking at a picture of a face. This finding was suggested to be due to the motivational responses to gaze direction being activated in the context of enhanced self-awareness by the presence of another person. The present results, thus, provide direct evidence that eye contact and gaze aversion between two persons influence the neural mechanisms regulating basic motivational-emotional responses and differentially activate the motivational approach-avoidance brain systems. PMID- 18402986 TI - Lipid rafts, cholesterol, and the brain. AB - Lipid rafts are specialized membrane microdomains that serve as organizing centers for assembly of signaling molecules, influence membrane fluidity and trafficking of membrane proteins, and regulate different cellular processes such as neurotransmission and receptor trafficking. In this article, we provide an overview of current methods for studying lipid rafts and models for how lipid rafts might form and function. Next, we propose a potential mechanism for regulating lipid rafts in the brain via local control of cholesterol biosynthesis by neurotrophins and their receptors. Finally, we discuss evidence that altered cholesterol metabolism and/or lipid rafts play a critical role in the pathophysiology of multiple CNS disorders, including Smith-Lemli-Opitz syndrome, Huntington's, Alzheimer's, and Niemann-Pick Type C diseases. PMID- 18402989 TI - Perceptual difficulty in source memory encoding and retrieval: prefrontal versus parietal electrical brain activity. AB - It is well established that source memory retrieval--remembering relationships between a core item and some additional attribute of an event--engages prefrontal cortex (PFC) more than simple item memory. In event-related potentials (ERPs), this is manifest in a late-onset difference over PFC between studied items which mandate retrieval of a second attribute, and unstudied items which can be immediately rejected. Although some sorts of attribute conjunctions are easier to remember than others, the role of source retrieval difficulty on prefrontal activity has received little attention. We examined memory for conjunctions of object shape and color when color was an integral part of the depicted object, and when monochrome objects were surrounded by colored frames. Source accuracy was reliably worse when shape and color were spatially separated, but prefrontal activity did not vary across the object-color and frame-color conditions. The insensitivity of prefrontal ERPs to this perceptual manipulation of difficulty stands in contrast to their sensitivity to encoding task: deliberate voluntary effort to integrate objects and colors during encoding reduced prefrontal activity during retrieval, but perceptual organization of stimuli did not. The amplitudes of ERPs over parietal cortex were larger for frame-color than object color stimuli during both study and test phases of the memory task. Individual variability in parietal ERPs was strongly correlated with memory accuracy, which we suggest reflects a contribution of visual working memory to long-term memory. We discuss multiple bottlenecks for source memory performance. PMID- 18402991 TI - Driver fatigue and highway driving: a simulator study. AB - Long duration of driving is a significant cause of fatigue-related accidents on motorways or major roadways. The fatigue caused by driving for extended periods acutely impairs driver alertness and performance and can compromise transportation safety. This study quantitatively measured the progression of driver fatigue and identified the conservative safe duration of continuous highway driving. Thirty young male subjects were analyzed during 90 min of laboratory-simulated highway driving. Sleepiness ratings (SSS) and reaction time (RT) tests were used to assess impairment of driver alertness and vigilance. Additionally, various measures of driving performance recorded throughout the experiment were used to measure temporal deterioration of driver performance from alert to fatigued using principal component analysis (PCA). The analytical results revealed that SSS scores, reaction times (RTs) and unstable driving performance significantly increased over time, indicating that excessive driving time is a significant fatigue factor and potential cause of fatigue-related accidents. Moreover, the analytical results indicated that 80 min was the safe limit for monotonous highway driving. Based on the experimental findings of this study, public awareness of the adverse affects of driver fatigue during long distance driving should be enhanced. This study provides explicit information of fatigue development that can be used to prevent fatigue-related accidents. PMID- 18402990 TI - Parietal contributions to recollection: electrophysiological evidence from aging and patients with parietal lesions. AB - There has been much recent investigation into the role of parietal cortex in memory retrieval. Proposed hypotheses include attention to internal memorial representations, an episodic working memory-type buffer, and an accumulator of retrieved memorial information. The current investigation used event-related potentials (ERPs) to test the episodic buffer hypothesis, and to assess the memorial contribution of parietal cortex in younger and older adults, and in patients with circumscribed lateral parietal lesions. In a standard recognition memory paradigm, subjects studied color pictures of common objects. One-third of the test items were presented in the same viewpoint as the study phase, one-third were presented in a 90 degrees rotated viewpoint, and one-third were presented in a noncanonical viewpoint. Conflicting with the episodic buffer hypothesis, results revealed that the duration of the parietal old/new effect was longest for the canonical condition and shortest for the noncanonical condition. Results also revealed that older adults demonstrated a diminished parietal old/new effect relative to younger adults. Consistent with previous data reported by Simons et al., patients with lateral parietal lesions showed no behavioral impairment compared to controls. Behavioral and ERP data from parietal lesion patients are presented and discussed. From these results, the authors speculate that the parietal old/new effect may be the neural correlate of an individual's subjective recollective experience. PMID- 18402992 TI - Isolation and functional analysis of two Cistus creticus cDNAs encoding geranylgeranyl diphosphate synthase. AB - Cistus creticus ssp. creticus is an indigenous shrub of the Mediterranean area. The glandular trichomes covering its leaf surfaces secrete a resin called "ladanum", which among others contains a number of specific labdane-type diterpenes that exhibit antibacterial and antifungal action as well as in vitro and in vivo cytotoxic and cytostatic activity against human cancer cell lines. In view of the properties and possible future exploitation of these metabolites, it was deemed necessary to study the geranylgeranyl diphosphate synthase enzyme (GGDPS, EC 2.5.1.30), a short chain prenyltransferase responsible for the synthesis of the precursor molecule of all diterpenes. In this work, we present the cloning, functional characterisation and expression profile at the gene and protein levels of two differentially expressed C. creticus full-length cDNAs, CcGGDPS1 and CcGGDPS2. Heterologous yeast cell expression system showed that these cDNAs exhibited GGDPS enzyme activity. Gene and protein expression analyses suggest that this enzyme is developmentally and tissue-regulated showing maximum expression in trichomes and smallest leaves (0.5-1.0cm). This work is the first attempt to study the terpenoid biosynthesis at the molecular level in C. creticus ssp. creticus. PMID- 18402993 TI - Antiectoparasitic activity of the gum resin, gum haggar, from the East African plant, Commiphora holtziana. AB - The mechanism of ixodid tick (Acari: Ixodidae) repellency by gum haggar, a resin produced by Commiphora holtziana (Burseraceae), was investigated by evaluating activity against the cattle tick, Boophilus microplus. In an arena bioassay, a hexane extract of the resin of C. holtziana exhibited a repellent effect lasting up to 5h. The hydrocarbon fraction of the resin extract was shown to account for the repellent activity, and was analysed by coupled gas chromatography-mass spectrometry (GC-MS). Major sesquiterpene hydrocarbons were tentatively identified as germacrene-D, delta-elemene and beta-bourbonene. The identity and stereochemistry of the former compound was confirmed as the (+)-isomer by peak enhancement using enantioselective GC, whereas the latter 2 compounds, which are most likely degradation products of germacrene-type precursors, were identified through isolation by preparative gas chromatography followed by microprobe-NMR spectroscopy. GC comparison of gum haggar with another resin, C. myrrha, which was inactive in the tick bioassay, showed that the latter contained much lower levels of these hydrocarbons. To assess the suitability of the gum haggar resin as a general acarine repellent, further tests were made on a major acarine pest of European and US animal husbandry systems, the red poultry mite, Dermanyssus gallinae (Acari: Dermanyssidae). Gum haggar extract, and the isolated hydrocarbon fraction, showed strong repellent effects in an olfactometer assay, and again gum myrrh showed no effect. These findings provide a scientific basis for the observed anti-tick properties of gum haggar, and demonstrate the potential for its development as a general acarine repellent for use in animal husbandry systems. PMID- 18402994 TI - [An humanitarian mission]. PMID- 18402995 TI - Economic burden of dengue infections in India. AB - Dengue infections are a significant cause of morbidity and mortality and lead to adverse economic effects in many developing tropical countries. In this study, we estimated the economic burden faced by India during the 2006 dengue epidemic. Costs incurred in managing a cohort of serologically confirmed dengue patients at a tertiary-level private hospital in north India were calculated. The median cost of treatment per hospitalised dengue patient was US$432.2 (95% CI US$343.6-625). A sensitivity analysis was performed to estimate the costs to the health system in India using a regression model. Variables with potentially large variations, including the ratio of unreported to reported cases and of hospitalised to ambulatory cases, the proportion requiring transfusions, loss of economic activities due to loss of workdays and deaths, were used. The average total economic burden was estimated to be US$27.4 million (95% CI US$25.7-29.1 million). Costs in the private heath sector were estimated to be almost four times public sector expenditures. Considerable economic losses (at a macro level) are incurred by developing countries like India during each dengue epidemic. Accurate estimates of the proportions of reported to unreported and of hospitalised to ambulatory dengue cases in India are needed to refine further the estimates of financial burden due to dengue in India. PMID- 18402996 TI - Ultrastructural damage to heart tissue from repeated oral exposure to yessotoxin resolves in 3 months. AB - Yessotoxin (YTX), an algal toxin contaminating edible shellfish, was previously shown to induce ultrastructural changes in some cardiac muscle cells of mice after acute (1 and 2mg/kg) or daily repeated oral exposure (1 and 2mg/kg/day, for 7 days). Therefore, the temporal evolution of the ultrastructural myocardial alterations and the development of other signs of toxicity induced by a repeated daily oral administration of YTX (1mg/kg/day, for 7 days) to mice were evaluated within 3 months after the treatment. Symptoms, food consumption, body weight, gross pathology and histopathology of the main organs and tissues were observed, and plasma levels of transaminases, lactate dehydrogenase, creatinine and creatinine phosphokinase were measured. Heart, liver, kidneys and cerebellum were also analysed by transmission electron microscopy. In addition, the blood concentration of YTX was determined by a direct enzyme linked immunosorbent assay (ELISA) 24h after the last toxin administration. No mortality or other treatment related changes, including histological or hematoclinical parameters, were recorded in mice administered with YTX. Similarly, electron microscopy did not reveal any ultrastructural alteration in the liver, kidneys, and cerebellum associated with YTX treatment. In contrast, changes in cardiac muscle cells near to the capillaries (clusters of rounded mitochondria and disorganization of myofibrils) were observed 24h after the treatment. These changes were also noted 30 days after the toxin administration, while after 90 days no differences in cardiac muscle cells between control and YTX-treated mice were observed, which indicated a recovery of the ultrastructural alterations induced by the toxin. PMID- 18402998 TI - Sensitivity to second-order motion in 10-month-olds. AB - Ten-month-old infants' sensitivity to first-order motion (FOM) defined by luminance and second-order motion (SOM) defined by flickering was measured in an eye-tracking paradigm. We used a small single disc or gratings moving horizontally. Although infants could track the SOM of a small disc, they failed to exhibit smooth pursuit eye movements. They also failed to track SOM gratings with smooth pursuit. However, the gain of tracking based on slow eye movement was influenced by the motion direction of SOM in cases when both FOM and SOM were presented simultaneously, suggesting some sensitivity to SOM. PMID- 18402997 TI - An SV40 VP1-derived epitope recognized by CD8+ T cells is naturally processed and presented by HLA-A*0201 and cross-reactive with human polyomavirus determinants. AB - The CD8+ T cell responses directed toward the VP1 antigens of human polyomaviruses JC and BK recently were shown to be cross-reactive. Two HLA-A0201 restricted determinants from each virus have been defined and include JCp100-108 (ILMWEAVTL) and BKp108-116 (LLMWEAVTV) as well as JCp36-44 (SITEVECFL) and BKp44 52 (AITEVECFL). We asked whether VP1 from the related SV40 contains similar HLA A0201-restricted determinants. In this study, we demonstrate that CD8+ T cells specific for SV40 VP1 p110-118 (ILMWEAVTV), but not p46-54 (SFTEVECFL), can be induced in HLA-A0201-transgenic mice and that these CD8+ T cells cross-react with the corresponding determinants from JC and BK virus. The SV40 p110 determinant was found to be processed and presented in SV40-infected cells. These results indicate that the JCp36/BKp44 determinants are distinctive for the human polyomaviruses while the JCp100/BKp108/SVp110 determinants are shared by all three viruses, providing a target for CD8+ T cell cross-reactivity. PMID- 18402999 TI - Biomagnification of organic pollutants in benthic and pelagic marine food chains from the Baltic Sea. AB - The trophic transfer of organic pollutants with varying physical chemical properties was determined in both a pelagic and benthic food chain using delta 15N as a continuous variable for assessing trophic levels. The trophic transfer of organic pollutants through the entire food chain in terms of food chain magnification factors (FCMFs) was quantified from the slope of the regression between ln [concentration] and delta 15N. Organic pollutants with statistically significant FCMFs >1 were considered to biomagnify within the food chain, whereas those with FCMFs <1 were considered to trophically dilute. Statistically significant FCMFs >1 were found for PCB congeners and organochlorine pesticides in the Baltic food chains whereas statistically significant FCMFs <1 were found for PAHs and PCNs due to trophic dilution resulting from metabolism. FCMFs were generally greater in the pelagic food chain than in the benthic food chain. However, estimated FCMFs for the benthic food chain are likely in error, as the delta 15N method suggested a food chain structure which was not consistent with the known dietary patterns of the species. Biomagnification factors (BMFs) were additionally calculated as the ratio of the lipid normalized concentrations in the predator and prey species with adjustment for trophic level and were generally consistent with the FCMFs with BMF >1 for PCBs and organochlorines. PMID- 18403000 TI - Spatial and temporal patterns of throughfall chemistry within a temperate mixed oak-beech stand. AB - The effects of meteorological variables and local canopy characteristics on throughfall chemistry were investigated within a mixed oak-beech stand during the leafed and the leafless seasons. Throughfall was collected after each rain event in 12 structural units delimited by three neighbouring trees each. For each investigated ion (H+, Mg2+, Ca2+, K+, NH4+, Na+, Cl(-), NO3(-) and SO4(2-)), mixed linear models were used to analyse the effects of the rain volume (R) and of the length of the preceding dry period (ADP) on net throughfall ion fluxes and to evaluate the influence of canopy density and tree species composition on these relationships. The models explained more than 70% of the total variability. Canopy characteristics accounted for most of the spatial variability while a large part of the temporal variability was not explained by R and ADP, suggesting an influence of other variables such as wind force and direction. For each season, net throughfall ion fluxes generally increased with increasing R and ADP, whose coefficients were interpreted as leaching capacity and dry deposition rate, respectively; H+ is an exception with negative exchange capacity (i.e. absorption) during the leafed season. Dry deposition rate and exchange capacity estimates were generally lower for the leafless canopies compared with the leafed season. For a given density level, exchange capacities were either higher for oak than for beech or similar for both species while dry deposition tended to increase with increasing beech proportion. The enhancing effect of canopy density on dry deposition was particularly marked for the mixed leafed canopies. For the pure canopies, the effect of canopy density on dry deposition rate and exchange capacity tended to be more pronounced during the leafless period compared with the leafed season, while it was generally limited compared with the species effect. PMID- 18403001 TI - Left ventricular apical torsion and architecture are not inverted in situs inversus totalis. AB - Occasionally, individuals have a complete, mirror-image reversal of their internal organ position, called situs inversus totalis (SIT). Whereas gross anatomy is mirror-imaged in SIT, this might not be the case for the internal architecture of organs, e.g. the myofiber pattern in the left cardiac ventricle. We performed a Magnetic Resonance Tagging study in nine controls and in eight subjects with SIT to assess the deformation pattern in the apical half of the LV wall. It appeared that both groups had the same LV apical deformation pattern. This implies that not only the superficial LV apical layers in SIT follow a normal, not inverted pattern, but the deeper layers as well. Apparently, the embryonic L/R controlling genetic pathway does determine situs-specific gross anatomy morphogenesis but it is not the only factor regulating fiber architecture within the apical part of the LV wall. We propose that mechanical forces generated in the not-inverted molecular structure of the basic right-handed helical contractile components of the sarcomere play a role in shaping the LV apex. PMID- 18403002 TI - Locomotor stimulation produced by 3,4-methylenedioxymethamphetamine (MDMA) is correlated with dialysate levels of serotonin and dopamine in rat brain. AB - (+/-)-3,4-Methylenedioxymethamphetamine (MDMA, or Ecstasy) is an illicit drug that evokes transporter-mediated release of monoamines, including serotonin (5 HT) and dopamine (DA). Here we monitored the effects of MDMA on neurochemistry and motor activity in rats, as a means to evaluate relationships between 5-HT, DA, and behavior. Male rats undergoing in vivo microdialysis were housed in chambers equipped with photobeams for measurement of ambulation (i.e., forward locomotion) and stereotypy (i.e., head weaving and forepaw treading). Microdialysis probes were placed into the n. accumbens, striatum or prefrontal cortex in separate groups of rats. Dialysate samples were assayed for 5-HT and DA by microbore HPLC-ECD. Rats received two i.v. injections of MDMA, 1 mg/kg followed by 3 mg/kg 60 min later; neurochemical and locomotor parameters were measured concurrently. MDMA produced dose-related elevations in extracellular 5 HT and DA in all regions, with the magnitude of 5-HT release always exceeding that of DA release. MDMA-induced ambulation was positively correlated with dialysate DA levels in all regions (P<0.05-0.0001) and with dialysate 5-HT in striatum and cortex (P<0.001-0.0001). Stereotypy was strongly correlated with dialysate 5-HT in all areas (P<0.001-0.0001) and with dialysate DA in accumbens and striatum (P<0.001-0.0001). These data support previous work and suggest the complex spectrum of behaviors produced by MDMA involves 5-HT and DA in a region- and modality-specific manner. PMID- 18403003 TI - Computer-delivered interventions for health promotion and behavioral risk reduction: a meta-analysis of 75 randomized controlled trials, 1988-2007. AB - OBJECTIVE: The use of computers to promote healthy behavior is increasing. To evaluate the efficacy of these computer-delivered interventions, we conducted a meta-analysis of the published literature. METHOD: Studies examining health domains related to the leading health indicators outlined in Healthy People 2010 were selected. Data from 75 randomized controlled trials, published between 1988 and 2007, with 35,685 participants and 82 separate interventions were included. All studies were coded independently by two raters for study and participant characteristics, design and methodology, and intervention content. We calculated weighted mean effect sizes for theoretically-meaningful psychosocial and behavioral outcomes; moderator analyses determined the relation between study characteristics and the magnitude of effect sizes for heterogeneous outcomes. RESULTS: Compared with controls, participants who received a computer-delivered intervention improved several hypothesized antecedents of health behavior (knowledge, attitudes, intentions); intervention recipients also improved health behaviors (nutrition, tobacco use, substance use, safer sexual behavior, binge/purge behaviors) and general health maintenance. Several sample, study and intervention characteristics moderated the psychosocial and behavioral outcomes. CONCLUSION: Computer-delivered interventions can lead to improved behavioral health outcomes at first post-intervention assessment. Interventions evaluating outcomes at extended assessment periods are needed to evaluate the longer-term efficacy of computer-delivered interventions. PMID- 18403004 TI - The mating behavior and reproduction performance in a multi-sire mating system for pigs. AB - An important aim of organic animal production is to allow natural animal behaviour. Regarding reproduction techniques, artificial insemination is permitted but natural mating is preferred. The outdoor multi-sire system, where the sows are placed in large paddocks with a group of boars, is one example of a service system, which complies well with the organic ideals of facilitating natural animal behavior. However, very little knowledge is available about such system. Seven groups of in total of 47 sows and 31 boars were observed to study the mating behavior in an outdoor multi-sire mating system and the subsequent reproduction results. The time of start of courtship, behavior and the cause of disruption if the courtship was terminated, were recorded each time a boar courted a sow. All aggressive interactions between the boars were also recorded to estimate the boar ranking order. The observations revealed numerous poor quality matings, a huge variation in the number of times sows are mated, and overworked boars. Only 35% of all copulations lasted 2min or more and 63% of all copulations were disrupted, mainly by competitor boars. The higher social status of the boar, the more copulations did it disrupt (p<0.05). The outcome was an unacceptable variation in reproduction results. Only 71% of all estrus sows conceived, corresponding to a pregnancy rate of 77% of all mated sows. A large inter-group variation in reproduction performance was observed, indicating scope for improvements. In some groups all sows showed estrus and all sows conceived. Recommendations for improvement of the system are proposed. PMID- 18403005 TI - Ubiquitination and carbonylation as markers of oxidative-stress in Ruditapes decussatus. AB - Environmental pollutants, such as metals, are widespread in aquatic environments and can lead to the formation of reactive oxygen species (ROS). ROS are highly toxic in marine species since they can cause serious reversible and irreversible changes in proteins including ubiquitination and modifications such as carbonylation. This study aimed to confirm the potential of ubiquitination and carbonylation as markers of oxidative stress in the clam Ruditapes decussatus (Veneroida, Veneridae) exposed to cadmium (40 microg/L). After 21 days of exposure clams were dissected into gills and digestive gland. Cytosolic proteins of both tissues were separated by two-dimensional electrophoresis (2-D SDS-PAGE) and analysed by immunobloting. Higher ubiquitination and carbonylation levels were in digestive gland of contaminated organisms. These results confirm the potential of ubiquitination and carbonylation as a sensitive and specific marker of oxidative stress in marine bivalves. In this approach, changes in protein structure provide options for affinity selection of sub-proteomes for 2D SDS PAGE, simplifying the detection of protein biomarkers using proteomic approach. PMID- 18403006 TI - Decline of brown trout (Salmo trutta) in Switzerland - how to assess potential causes in a multi-factorial cause-effect relationship. AB - A considerable decline in fish catch in Switzerland triggered a 5-year project (Fischnetz) to investigate potential causes. The results of many field and laboratory studies, together with literature and historical data, indicated a need for a concise synthesis of the results. Therefore, Bayesian probability network (BPN) and weight-of-evidence (WOE) approaches were applied. Here, these both approaches are compared and evaluated. In addition, the potential reasons for the (mis)match in the two approaches were evaluated. In both studies, proliferative kidney disease (PKD), caused by a parasite, and the clinical outbreak supported by various factors, was a very probable single parameter. WOE assessed habitat and streambed quality as likely for contributing to impaired health, recruitment and abundance at single sites only, but this parameter was assessed to be the most important and ubiquitous stressor in the BPN. Mismatches suggested that these factors were either not considered or not equally assessed by the different models, which is due to different endpoints, incomplete data sets and different handling of these various data sets by the applied synthesis approaches. PMID- 18403007 TI - Exposure of sticklebacks (Gasterosteus aculeatus) to cadmium sulfide nanoparticles: biological effects and the importance of experimental design. AB - Effects of nanoparticles on aquatic organisms have been little studied to date and toxicological data are urgently needed for development of regulatory frameworks for these substances. Here, we report the findings of a study exposing sticklebacks to cadmium sulfide (CdS) as bulk material and quantum dots. Fish were exposed for 21 d in a flow through test system to 5, 50 or 500 microg l(-1) CdS nanoparticles (nCdS) coated in thiol terminated methyl polyethylene glycol (MPEG), bulk CdS or MPEG at 500 microg l(-1) (nominal concentrations). With the exception of the highest nCdS exposure, measured concentrations were approximately one order of magnitude below nominal. A single fish from each group (excluding MPEG) was examined using energy dispersive X-ray analysis (EDX) to localise cadmium, however, cadmium could not be detected in whole body sections. Elevated levels of oxidized glutathione were measured in the gills of fish exposed to 50 and 500 microg l(-1) nCdS. Induction of vitellogenin synthesis was not detected in any of the treatment groups. The number of males engaged in nest building behaviour following exposure to 500 microg l(-1) nCdS was reduced and livers of 4/6 fish in the same treatment displayed hepatocellular nuclear pleomorphism. The results are discussed emphasising the fundamental importance of experimental design and the need to understand the behaviour of nanoparticles in the aqueous phase. PMID- 18403008 TI - Evaluation of bioavailable hydrocarbon sources and their induction potential in Prince William Sound, Alaska. AB - To realistically evaluate the consequences of exposure to a complex mixture, we modified a passive sampler technology, the semipermeable membrane device (SPMD), which absorbs the bioavailable hydrophobic organic compounds present in an environment. These samplers were deployed in Prince William Sound (PWS), Alaska, at locations selected as potential sites of hydrocarbon deposition, as well as in random sites for regional assessment. Some of these sites were affected by previous human activity, such as canneries and salmon hatcheries, while others were sites of oil discharge as a consequence of the 1964 earthquake or the oil spill of T/V Exxon Valdez in 1989. The SPMDs were deployed for 27-28 d, processed, and then split, with one aliquot dedicated to chemical analysis and the other injected into juvenile rainbow trout (Oncorhynchus mykiss), along with the proper controls including a solvent control, field blank, and positive control. Trout fry were sacrificed after 2 or 7d, and their livers assayed for CYP1A induction by the standard bioassay for hydrocarbon exposure, the ethoxyresorufin-o-deethylase (EROD) assay. The results of this study were consistent and reproducible and showed that oil, whether deposited in 1964 or 1989, is still bioavailable as it can elicit as sustained response. Also, the same oil deposited in different sites of the same region has degraded differently, which is demonstrated by this method. Other putative sources of hydrocarbons, such as oil seeps, were dismissed as regional sources of induction agents as the responses following injection of modified SPMD extract from those sites did not differ significantly from the solvent control. This is a flexible, sensitive method that assesses the response to site-specific bioavailable contaminants and does so within the normal physiological response range of the target. PMID- 18403009 TI - Nuclear changes in blood, early liver anomalies and hepatocellular cancers in flounder (Platichthys flesus L.) as prognostic indicator for a higher cancer risk? AB - Nuclear changes (micronuclei) in blood are a well-established biomarker of toxic injury and related genotoxicity in mussel, fish and humans, as well. Yet, the relationship between the frequencies of micronuclei in blood and nuclear changes in liver cells itself and the higher potential risk to develop liver cancer is not yet fully implemented. We analysed and diagnosed macroscopic liver changes and H&E and Feulgen stained histological liver sections (2 microm of thickness) together with blood smears (stained with Sytox Green) from same individuals in flounder caught along a well-known contaminant gradient in the North Sea. Results showed an increase in micronuclei frequencies in blood at the more contaminated sites and higher frequencies of macroscopically visible neoplastic liver changes. In livers of the same individuals, nuclear changes were diagnosed in early to degenerative liver lesion, extra-focal tissue surrounding neoplastic lesion and various stages during cancer progression from early neoplastic foci, benign adenomas and malignant carcinomas. The aim of this approach is to expand diagnostic criteria, well-established in the frame of BEQUALM to nuclear changes which might help diagnosis of malignancies in doubtful cases. PMID- 18403010 TI - The liver of wrasse - morphology and function as a mirror of point source chemical impact. AB - Corkwing wrasse (Symphodus melops L.), a protogynous, non-migratory lipfish species, living close to rocky shores was chosen as an indicator species for the monitoring of biological effects of contaminants. Fish were caught by local fisherman at the Norwegian west coast in fjord sites within the framework of the EU BEEP project. The sites represented different point source impacts of (I) copper (a former copper mine), (II) polycyclic aromatic hydrocarbons (PAHs, aluminium smelter discharge), (III) formaldehyde plus PAHs (kelp-factory and influence of the aluminium smelter). Livers of wrasse were studied for histopathological alterations and compared to healthy livers of fish from a reference location. Besides liver morphology, different functional and metabolic parameters were measured to link pathological alterations to functional disorders. The integrity of the lysosomal compartment was tested by the assessment of lysosomal membrane stability (lys), and the accumulation of neutral lipids and lipofuscin. Activity and intracellular localisation of the NADPH producing enzymes in the liver were assessed histochemically and measured by computer-assisted image analysis. Histopathological alterations were most severe at the site impacted by formaldehyde and PAHs. These findings were associated with highest tumor prevalence, lowest membrane stabilities in hepatocytes and highest accumulation rates of lipofuscin in the liver. The activities of the NADPH-producing enzymes phosphogluconate dehydrogenase (PGDH) and glucose-6 phosphate dehydrogenase (G6PDH) were significantly lower compared to unimpacted reference fish. Histopathological alterations showed clear differences dependent on the input source. Potential links between specific contaminant impact and functional and morphological disorders are discussed. PMID- 18403011 TI - The slow resorption with replacement by bone of a hydrothermally synthesized pure calcium-deficient hydroxyapatite. AB - A newly developed calcium-deficient hydroxyapatite composed of rod-shaped particles synthesized by the hydrothermal method (HHA) and stoichiometric hydroxyapatite (SHA) synthesized by the sintering method was used for in vivo implantation and in vitro culture systems to compare these biological responses. In the rabbit femur, implanted HHA was slowly resorbed and about 80% of the implant remained 24 weeks after implantation; however, up to 72 weeks after implantation, most of the implanted HHA was resorbed. The implanted SHA was unresorbed throughout the experimental period, but degradation by the invasion of newly formed bone was seen at 72 weeks after implantation. Bone histomorphometry showed that the volume of newly formed bone and the number of osteoclasts in the implanted region were significantly higher in HHA than in SHA 24 weeks after implantation. In vitro culture of C2C12 cells with the induction of osteoblastic phenotypes using recombinant bone morphogenetic protein-2 showed similar cell density and the induction of alkaline phosphatase activity between the cells on HHA and SHA discs. In vitro osteoclastogenesis of HHA and SHA discs using bone marrow macrophages and recombinant receptor activator of nuclear factor-kappaB ligand showed higher TRAP activity of osteoclasts cultured on HHA discs. These results showed that slow biodegradability did not always correlate to final replaceability in bone tissue, and suggested that the activity of osteoclasts correlated to the bone-forming activity of osteoblasts. PMID- 18403012 TI - Three-dimensional electrospun ECM-based hybrid scaffolds for cardiovascular tissue engineering. AB - Electrospinning using natural proteins or synthetic polymers is a promising technique for the fabrication of fibrous scaffolds for various tissue engineering applications. However, one limitation of scaffolds electrospun from natural proteins is the need to cross-link with glutaraldehyde for stability, which has been postulated to lead to many complications in vivo including graft failure. In this study, we determined the characteristics of hybrid scaffolds composed of natural proteins including collagen and elastin, as well as gelatin, and the synthetic polymer poly(epsilon-caprolactone) (PCL), so to avoid chemical cross linking. Fiber size increased proportionally with increasing protein and polymer concentrations, whereas pore size decreased. Electrospun gelatin/PCL scaffolds showed a higher tensile strength when compared to collagen/elastin/PCL constructs. To determine the effects of pore size on cell attachment and migration, both hybrid scaffolds were seeded with adipose-derived stem cells. Scanning electron microscopy and nuclei staining of cell-seeded scaffolds demonstrated the complete cell attachment to the surfaces of both hybrid scaffolds, although cell migration into the scaffold was predominantly seen in the gelatin/PCL hybrid. The combination of natural proteins and synthetic polymers to create electrospun fibrous structures resulted in scaffolds with favorable mechanical and biological properties. PMID- 18403013 TI - Physical properties and cellular responses to crosslinkable poly(propylene fumarate)/hydroxyapatite nanocomposites. AB - A series of crosslinkable nanocomposites has been developed using hydroxyapatite (HA) nanoparticles and poly(propylene fumarate) (PPF). PPF/HA nanocomposites with four different weight fractions of HA nanoparticles have been characterized in terms of thermal and mechanical properties. To assess surface chemistry of crosslinked PPF/HA nanocomposites, their hydrophilicity and capability of adsorbing proteins have been determined using static contact angle measurement and MicroBCA protein assay kit after incubation with 10% fetal bovine serum (FBS), respectively. In vitro cell studies have been performed using MC3T3-E1 mouse pre-osteoblast cells to investigate the ability of PPF/HA nanocomposites to support cell attachment, spreading, and proliferation after 1, 4, and 7 days. By adding HA nanoparticles to PPF, the mechanical properties of crosslinked PPF/HA nanocomposites have not been increased due to the initially high modulus of crosslinked PPF. However, hydrophilicity and serum protein adsorption on the surface of nanocomposites have been significantly increased, resulting in enhanced cell attachment, spreading, and proliferation after 4 days of cell seeding. These results indicate that crosslinkable PPF/HA nanocomposites are useful for hard tissue replacement because of excellent mechanical strength and osteoconductivity. PMID- 18403014 TI - Neuropilin-1 is expressed by chronic lymphocytic leukemia B cells. PMID- 18403015 TI - Phospholipase C, phosphatidylinositol 3-kinase, and intracellular [Ca(2+)] mediate the activation of chicken HD11 macrophage cells by CpG oligodeoxynucleotide. AB - The activation of phospholipases is one of the earliest key events in receptor mediated cellular responses to a number of extracellular signaling molecules. Oligodeoxynucleotides containing CpG motifs (CpG ODN) mimic microbial DNA and are immunostimulatory to most vertebrate species. In the present study, we used the production of nitric oxide (NO) as an indicator to evaluate the involvement of the signaling cascades of phospholipases and phosphatidylinositol 3-kinase (PI3K) in the activation of chicken HD11 macrophage cells by CpG ODN. Using selective inhibitors, we have identified the involvement of phosphatidylinositol (PI) phospholipase C (PI-PLC), but not phosphatidylcholine (PC)-phospholipase C (PC PLC) and PC-phospholipase D (PC-PLD), in CpG ODN-induced NO production in HD11 cells. Preincubation with PI-PLC selective inhibitors (U-73122) completely abrogated CpG ODN-induced NO production in HD11 cells, whereas PC-PLC inhibitor (D609) and PC-PLD inhibitor (n-butanol) had no inhibitory effects. Additionally, inhibition of PI3K and protein kinase C (PKC) with selective inhibitors and chelation of intracellular [Ca(2+)] also significantly attenuated NO production in CpG ODN-activated HD11 cells. Our results demonstrate that PI-PLC, PI3 K, PKC, and intracellular [Ca(2+)] are important components of the CpG ODN-induced signaling pathway that leads to the production of NO in avian macrophage cells. PMID- 18403017 TI - When the trivial becomes meaningful: reflections on a process evaluation of a home visitation programme in South Africa. AB - This paper reflects on a process evaluation of a home visitation programme in South Africa. The programme, implemented in two low-income communities, focused on the reduction of risks to unintentional childhood injuries. The evaluation comprised a combination of qualitative and quantitative methods, including observations in conjunction with an evaluator's journal, diaries kept by the home visitors, interviews and focus group discussions. Short questionnaires were administered to programme staff and home visitors. Caregivers were visited to attain their assessment of visitors and the programme. These methods resulted in a detailed description of implementation processes, but more importantly gave insight into the experiences and perceptions of the social actors, i.e. programme staff, visitors and caregivers. It also offered possible explanations for the difference in the intervention effect between the two sites. Two major challenges to the evaluation were: (i) the power-imbalance between the evaluator and community participants (visitors and caregivers) and (ii) the language- and cultural barriers between evaluator and community participants. The evaluation demonstrated that process information can contribute towards explaining outcome results, but also that active participation from all social actors is a necessary condition if process evaluations are to result in programme improvement. PMID- 18403016 TI - Optimization and application of glutamate cysteine ligase measurement in wildlife species. AB - Glutamate cysteine ligase (GCL), synthesizing gamma-glutamylcysteine from glutamate and cysteine, is the rate-limiting enzyme in glutathione (GSH) biosynthesis. GCL activity measurement was optimized in tissues from deer mice, Sprague Dawley rats, and mallard ducks. Varying glutamic acid concentrations from 5 to 80 mM did not affect GCL activities markedly, whereas cysteine concentrations from 2.5 to 40 mM influenced GCL activities substantially. Optimal cysteine concentrations for deer mouse, Sprague Dawley rat, and mallard duck (respectively) were 30, 30, and 20 mM in liver, 10, 10, and 20 mM in kidney, 20, 20, and 30 mM in brain, and 30 mM in heart for all three species. Responses of mallard duck GCL activity to acid metalliferous water were evaluated. After subacute exposure, low doses increased GCL activity and GSH content in liver by 48.3% and 54.4%, respectively. High doses reduced GCL activities significantly in liver and kidney to 31.2% and 43.0% of the control, respectively. PMID- 18403018 TI - Methodological and conceptual issues confronting a cross-country Delphi study of educational program evaluation. AB - Although the Delphi is widely used, research on certain methodological issues is somewhat limited. After a brief introduction to the strengths, limitations, and methodological challenges of the technique, we share our experiences (as well as problems encountered) with an electronic Delphi of educational program evaluation (EPE) in the Asia-Pacific region. The study is described followed by a discussion of the difficulties in participant recruitment and selection, sample size, instrumentation, data collection and analysis and attempts to resolve them. Some of these problems are generic to the Delphi whereas others related to the specifics of the investigation. What we learned should be useful for future Delphis with a similar focus. PMID- 18403019 TI - A CD4 homologue in sea bass (Dicentrarchus labrax): molecular characterization and structural analysis. AB - CD4 is a transmembrane glycoprotein fundamental for cell-mediated immunity. Its action as a T cell co-receptor increases the avidity of association between a T cell and an antigen-presenting cell by interacting with portions of the complex between MHC class II and TR molecules. In this paper we report the cDNA cloning, expression and structural analysis of a CD4 homologue from sea bass (Dicentrarchus labrax). The sea bass CD4 cDNA consists of 2071 bp that translates in one reading frame to give the entire molecule containing 480 amino acids. The analysis of the sequence shows the presence of four putative Ig-like domains and that some fundamental structural features, like a disulphide bond in domain D2 and the CXC signalling motif in the cytoplasmic tail, are conserved from sea bass to mammals. Real-time PCR analysis showed that very high levels of CD4 mRNA transcripts are present in thymus, followed by gut and gills. In vitro stimulation of head kidney leukocytes with LPS and PHA-L gave an increase of CD4 mRNA levels after 4h and a decrease after 24h. Homology modelling has been applied to create a 3D model of sea bass CD4 and to investigate its interaction with sea bass MHC-II. The analysis of the 3D complex between sea bass CD4 and sea bass MHC-II suggests that the absence of a disulfide bond in the CD4 D1 domain could make this molecule more flexible, inducing a different conformation and affecting the binding and the way of interaction between CD4 and MHC-II. Our results will add new insights into the sea bass T cell immune responses and will help in the identification of T cell subsets in teleost fishes to better understand the evolution of cell-mediated immunity from fish to mammals. PMID- 18403020 TI - Role of STAT3 in glucocorticoid-induced expression of the human IL-10 gene. AB - In the present report we have determined the molecular mechanisms, which govern the expression of the human IL-10 gene when induced by the glucocorticoid Methyl Prednisolone (MP). Treatment of cells with MP at 10(-6) M will readily induce IL 10 in CD19+ primary B cells and in a human B cell line. Analysis of the IL-10 promoter showed a robust 18-fold induction and demonstrated that a potential GRE motif was not required, while mutation of the -120 STAT-motif strongly reduced MP induced trans-activation. A strong induction was also seen with a trimeric STAT motif and over-expression of dominant-negative STAT3 could block MP induction of IL-10 mRNA. Finally, MP treatment induced binding of STAT3 to the promoter as shown by gelshift, supershift and by chromatin-immunoprecipitation. These data show that glucocorticoid-induced expression of the IL-10 gene is mediated by the transcription factor STAT3. PMID- 18403021 TI - Developmental neurotoxicity testing in vitro: models for assessing chemical effects on neurite outgrowth. AB - In vitro models may be useful for the rapid toxicological screening of large numbers of chemicals for their potential to produce toxicity. Such screening could facilitate prioritization of resources needed for in vivo toxicity testing towards those chemicals most likely to result in adverse health effects. Cell cultures derived from nervous system tissue have proven to be powerful tools for elucidating cellular and molecular mechanisms of nervous system development and function, and have been used to understand the mechanism of action of neurotoxic chemicals. Recently, it has been suggested that in vitro models could be used to screen for chemical effects on critical cellular events of neurodevelopment, including differentiation and neurite growth. This review examines the use of neuronal cell cultures as an in vitro model of neurite outgrowth. Examples of the cell culture systems that are commonly used to examine the effects of chemicals on neurite outgrowth are provided, along with a description of the methods used to quantify this neurodevelopmental process in vitro. Issues relating to the relevance of the methods and models currently used to assess neurite outgrowth are discussed in the context of hazard identification and chemical screening. To demonstrate the utility of in vitro models of neurite outgrowth for the evaluation of large numbers of chemicals, efforts should be made to: (1) develop a set of reference chemicals that can be used as positive and negative controls for comparing neurite outgrowth between model systems, (2) focus on cell cultures of human origin, with emphasis on the emerging area of neural progenitor cells, and (3) use high-throughput methods to quantify endpoints of neurite outgrowth. PMID- 18403022 TI - Two instances of large genome profile picobirnavirus occurrence in Argentinian infants with diarrhea over a 26-year period (1977-2002). AB - BACKGROUND: Picobirnavirus' (PBV) association with diarrhea in children is not reliably established and the potential role of pathogenic PBV needs further investigations. OBJECTIVE: The aim of this study is to clarify the role of PBV in diarrhea illness in children. METHODS: Between January 1977 and December 2002, 2224 stool specimens were collected from children <3 years old with diarrhea illness. All samples were analyzed by the polyacrylamide gel electrophoresis technique (PAGE) for the presence of bisegmented dsRNA virus genomic pattern. Gels were dried and archived. This study procedure allowed us to keep a laboratory electrophoretic record of each sample assayed. In the present study, all the electrophoretic records were reviewed in order to identify PBV positive samples. RESULTS: Two out of 2224 (0.09%) stools were positive for large genome profile of PBV. These two positive samples were collected from hospitalized children <1 year old; one of them presenting rotavirus co-infection. CONCLUSIONS: The findings obtained in the present report support strong evidence that large genome profile PBV can be considered more an occasional viral agent rather than an etiological agent associated with diarrheal illness. PMID- 18403023 TI - Toll-like receptor genes are differentially expressed at the sites of infection during the progression of Johne's disease in outbred sheep. AB - Toll-like receptors (TLR) are engaged by ligands on microbial pathogens to initiate innate and adaptive immune responses. Little is known about TLR involvement during infection with Mycobacterium avium subsp. paratuberculosis (M. ptb), the cause of Johne's disease in ruminants, although there is a profound immunopathological response in affected animals. We have analyzed the expression of 10 TLR genes relative to validated reference genes at predilection sites in ileum, jejunum and associated lymph nodes as well as in peripheral blood, to determine if TLR expression is altered in response to infection with M. ptb in outbred sheep. Previously unexposed animals from two flocks and animals from three naturally infected flocks were used with restricted maximum likelihood linear mixed modeling applied to determine significant differences. These were related to the pathologies observed at different stages of infection in exposed sheep, after allowing for other sources of variation. In most cases there were differences in TLR expression between early paucibacillary and multibacillary groups when compared to uninfected sheep, with most TLRs for the paucibacillary group having lower expression levels than the multibacillary group. Increased expression of TLR1-5, and 8 was observed in ileum or jejunum, and TLR1-4, 6, and 8 in mesenteric lymph nodes. There was a trend for increased expression of TLR1, 2, and 6-8 in PBMCs of exposed compared to non-exposed animals. Further study of TLR expression in Johne's disease in ruminants is warranted as these observed differences may help explain pathogenesis and may be useful in the future diagnosis of M. ptb infection. PMID- 18403024 TI - Differential mediator production by dendritic cells upon toll-like receptor stimulation does not impact T cell cytokine expression. AB - Dendritic cells are key components of successful immunological responses bridging innate and adaptive defenses. In this study we wanted to know whether ligation of toll-like receptors (TLR) expressed by dendritic cells would induce differential proinflammatory mediator expression and whether these dendritic cells would differentially impact T cell function. For this purpose bone marrow-derived dendritic cells from OTII mice were used. The dendritic cells showed detectable levels of TLR1, 2, 4, 6, 7, 8 and 9, with TLR2 and TLR4 expressed at the highest levels. To determine whether TLR ligation differentially influenced proinflammatory mediator expression the dendritic cells were stimulated with peptidoglycan (PGN) or lipopolysaccharide (LPS) for TLR2 or TLR4, respectively. Comparisons were made to dendritic cells exposed to TNF-alpha or saline as controls. Whereas, both LPS and PGN were equally effective at inducing CXCL1 and TNF-alpha expression from the dendritic cells, LPS was unique at inducing CCL2 expression, and PGN was unique at inducing IL-1beta expression. Despite these differences, LPS and PGN treated dendritic cells were equally effective at eliciting IFN-gamma expression from T cells in an antigen-specific manner. These data indicate that ligation of TLR by components of Gram+ and Gram- bacteria differentially influence dendritic cell proinflammatory mediator expression, and that differential mediator production by dendritic cells upon TLR stimulation does not impact T cell cytokine production. PMID- 18403025 TI - Increased IL-23 secretion and altered chemokine production by dendritic cells upon CD46 activation in patients with multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS). In MS, myeloid dendritic cells (mDCs) secrete elevated amounts of IL-23, a potent proinflammatory cytokine, compared to healthy donors. Here, we examined the role of CD46, a complement binding factor, in mDCs by analyzing cytokine and chemokine production in healthy donors and patients with MS. There were striking differences between these groups with increased IL-23p19, CCL3 and CCL5 production, but decreased CCL2 levels in patients. This demonstrates major differences of DC activation upon CD46 activation, with a potential role in the pathogenesis of MS. PMID- 18403026 TI - Cochlear implantation in infants less than 12 months of age. AB - OBJECTIVE: Infants are diagnosed with severe to profound hearing loss at an earlier age due to the advent of universal newborn hearing screening. This offers the opportunity to provide intervention in the form of cochlear implantation at an earlier age than was previously possible. The purpose of this investigation is to evaluate the risk of cochlear implant surgery in children less than 12 months of age. DESIGN: Retrospective review of children who underwent cochlear implantation before 12 months of age. SETTING: Patients were identified from a database of pediatric cochlear implant patients at a tertiary care center. All patients were diagnosed with severe to profound hearing loss by otoacoustic emission and auditory brainstem response. Follow-up ranged from 2 months to 5 years. RESULTS: Fourteen of 15 patients had full insertions of the electrode hardware. Less than full insertion and post-operative CSF otorrhea occurred in one patient with severe cochlear abnormalities. There were no other perioperative surgical complications. The average speech detection threshold was 27.6 dB (20-45 dB) at approximately 1-3 months post-stimulation and 25 dB (15-30 dB) at approximately 5-7 months. CONCLUSION: In our experience, we feel cochlear implantation is safe for infants as young as 6 months of age. The current standard at our institution is to implant by 7 months of age for prelingual deafness as opposed to waiting additional time until 12 months of age before the brain is presented with speech. PMID- 18403027 TI - Characterization of polyclonal antibodies against the capsid proteins of Ljungan virus. AB - Ljungan virus (LV) is a suspected human pathogen isolated from voles in Sweden and North America. To enable virus detection and studies of localization and activity of virion proteins, polyclonal antibodies were produced against bacterially expressed capsid proteins of the LV strain, 87-012G. Specific detection of proteins corresponding to viral antigens in lysates of LV infected cells was demonstrated by immunoblotting using each one of the generated polyclonal antibodies. In addition, native viral antigens present in cell culture infected with LV strains 87-012G or 145SLG were detected in ELISA and by immunofluorescence using the antibodies against the VP0 and VP1 proteins. The anti-VP3 antibody did not react with native proteins of the LV virion, suggesting that the VP3 is less potent in evoking humoral response and may have a less exposed orientation in the virus capsid. No activity of the antibodies was observed against the closely related human parechovirus type 1. The polyclonal antibody against the VP1 protein was further used for detection of LV infected myocytes in a mouse model of LV-induced myocarditis. Thus, polyclonal antibodies against recombinant viral capsid proteins enabled detection of natural LV virions by several different immunological methods. PMID- 18403028 TI - SV40 reporter viruses. AB - Three simian virus 40 (SV40) reporter viruses were constructed in this study. One expresses the green fluorescent protein (GFP) as a fusion protein with the first exon of large-T (LT) antigen and is useful for live-cell imaging. A second reporter virus has a FLAG epitope tag at the C-terminus of large-T antigen (vC LT(FLAG)), and a third has the FLAG tag at the N-terminus of LT (vN-LT(FLAG)). The vC-LT(FLAG) construct grows to titers near those of wild-type (WT) virus and functions well as a reporter virus for SV40 infection. The vN-LT(FLAG) construct, while viable, has a defect in the production and spread of infectious particles. All three viruses are useful in detecting superinfecting virus in cells in which nuclear LT is already present, such as persistently infected human mesothelial cells. PMID- 18403029 TI - Gephyrin: where do we stand, where do we go? AB - Gephyrin is a multifunctional protein responsible for molybdenum cofactor synthesis and the clustering of glycine and GABA(A) receptors at inhibitory synapses. Based on the structure of its two conserved domains, G and E, gephyrin is thought to form a hexagonal lattice serving as a scaffold for accessory proteins at postsynaptic sites. However, important aspects of gephyrin gene expression, protein structure and regulation, as well as the role of gephyrin in synapse formation and plasticity, remain poorly understood. Here we review the current state of knowledge about gephyrin, highlighting new research avenues based on a different structural model and a revised nomenclature for gephyrin splice variants. Unraveling the biology of gephyrin will further our understanding of glycinergic and GABAergic synapses in health and disease. PMID- 18403030 TI - The mitochondrial impairment, oxidative stress and neurodegeneration connection: reality or just an attractive hypothesis? AB - Aging is the most important risk factor for common neurodegenerative disorders such as Parkinson's and Alzheimer's diseases. Aging in the central nervous system has been associated with elevated mutation load in mitochondrial DNA, defects in mitochondrial respiration and increased oxidative damage. These observations support a 'vicious cycle' theory which states that there is a feedback mechanism connecting these events in aging and age-associated neurodegeneration. Despite being an extremely attractive hypothesis, the bulk of the evidence supporting the mitochondrial vicious cycle model comes from pharmacological experiments in which the modes of mitochondrial enzyme inhibition are far from those observed in real life. Furthermore, recent in vivo evidence does not support this model. In this review, we focus on the relationship among the components of the putative vicious cycle, with particular emphasis on the role of mitochondrial defects on oxidative stress. PMID- 18403032 TI - Various optical coherence tomographic findings in restenotic lesions after sirolimus-eluting stent implantation. AB - In-stent restenosis (ISR) after sirolimus-eluting stent (SES) implantation rarely occurs, and these lesions are commonly recognized as focal restenosis on angiograms. Optical coherence tomography (OCT) is able to distinguish from several components of intracoronary structures by different signal patterns. We herein report three cases of various OCT images from ISR lesions in spite of similar angiographic findings. Our OCT findings suggested that ISR lesions that develop after SES implantation might be composed of not uniform but various tissues. PMID- 18403031 TI - Gap junctions: multifaceted regulators of embryonic cortical development. AB - The morphological development of the cerebral cortex from a primitive neuroepithelium into a complex laminar structure underlying higher cognition must rely on a network of intercellular signaling. Gap junctions are widely expressed during embryonic development and provide a means of cell-cell contact and communication. We review the roles of gap junctions in regulating the proliferation of neural progenitors as well as the migration and differentiation of young neurons in the embryonic cerebral cortex. There is substantial evidence that although gap junctions act in the classical manner coupling neural progenitors, they also act as hemichannels mediating the spread of calcium waves across progenitor cell populations and as adhesive molecules aiding neuronal migration. Gap junctions are thus emerging as multifaceted regulators of cortical development playing diverse roles in intercellular communication. PMID- 18403033 TI - Interface of epidemiology, pet population issues and policy. AB - Progress has been made towards eliminating Rowan's "statistical black hole" regarding pet populations and their dynamics, although deficiencies remain. The challenge in the coming decade will be to continue the progress, and generate meaningful epidemiologic data. Veterinary epidemiologists will continue to play an important role in the generation, dissemination and translation of results from their studies into coherent, effective companion animal welfare policy. PMID- 18403034 TI - Schwabe Symposia 2004-2006. Forward. PMID- 18403035 TI - Turning promiscuous kinase inhibitors into safer drugs. AB - Burgeoning interest in multi-target drugs to treat complex diseases and malignancies has motivated a reassessment of the therapeutic value of promiscuity. Although drug efficacy might not correlate with specificity, it would be risky to welcome promiscuous compounds without a rational strategy to control therapeutic impact. This situation might change as novel selectivity filters are incorporated into drug design. For example, cardiotoxic side effects of the cancer drug imatinib might be curbed by applying such premises. Here, we survey approaches to control the therapeutic impact of cross-reactive kinase inhibitors and advocate the application of a novel selectivity filter by illustrating its cleaning efficacy. Finally, we evaluate the possibility of turning multi-target kinase inhibitors into clinical opportunities through judicious chemical modifications. PMID- 18403036 TI - Effect of water activity on inactivation of Listeria monocytogenes and lactate dehydrogenase during high pressure processing. AB - The aim of this study was to investigate the effect of water activity (aw) on the inactivation of Listeria monocytogenes and lactate dehydrogenase (LDH) during high pressure processing (HPP). For microbial inactivation lyophilized cells of L. monocytogenes 19,115 were left dry or were suspended in 10 ml of 0.1% peptone water, 10 ml of glycerol, or mixtures of glycerol and peptone water. All samples of various aws were high pressure (HP) processed at ambient temperature at 600 MPa for 300 s. Following HPP, samples were serially diluted in 0.1% peptone and spread-plated on Tryptic Soy agar supplemented with Yeast Extract. For enzyme inactivation, 4.2 mg of lyophilized LDH was suspended in 2 ml of 100 mM phosphate buffer (pH 7.4), 2 ml of peptone water or glycerol, or in 2 ml mixtures of glycerol and peptone water. A lyophilized sample with no added liquid was also included. All enzyme samples were subjected to HPP as described above. After HPP, LDH was diluted to 0.28 microg/ml in 100 mM phosphate buffer (pH 7.4). LDH activity was assessed by measuring the change in concentration of beta-NADH as a function of time. Dynamic light scattering analysis (DLS) was performed to examine the size distribution, polydispersity, and hydrodynamic radius of LDH before and after HPP. No significant difference in CFU/g was observed between lyophilized cells not subjected to HPP and lyophilized cells subjected to 600 MPa for 300 s (P<0.05). However, lyophilized cells that were suspended in 100% to 60% peptone water showed a approximately 7.5-log(10) reduction when subjected to HPP. Survival of L. monocytogenes following HPP significantly increased (P<0.05) when the peptone water concentration was decreased below 60% (aw approximately 0.8). DLS results revealed that LDH suspended in buffer underwent aggregation following HPP (600 MPa, 300 s). Inactivation rate constants obtained using a first-order kinetic model indicated that untreated and HP processed lyophilized LDH had similar activities. When LDH was subject to HPP in solutions containing glycerol, enzyme activity decreased as the water content increased (r2=0.95). Lyophilization completely protected L. monocytogenes and LDH from inactivation by high pressure. Furthermore, enzyme activity and cell survival increased as water activity was decreased. We postulate low aw results in protein stabilization, which prevents protein denaturation and cell death during HPP. PMID- 18403037 TI - Distribution and composition of extracellular polymeric substances in membrane aerated biofilm. AB - Extracellular polymeric substances (EPS) are one of the main components of the biofilm and perform important functions in the biofilm system. In this study, two membrane-aerated biofilms (MABs) were developed for the thin and thick biofilms under different surface loading rates (SLRs). Supplies of oxygen and substrates in the MAB were from two opposite directions. This counter diffusion of nutrients resulted in a different growth environment, in contrast to conventional biofilms receiving both oxygen and substrates from the same side. The compositions, distributions and physicochemical properties (solubility and bindability) of EPS in the MABs of different thicknesses under different SLRs were studied. The effect of dissolved oxygen (DO) concentration within the MAB on EPS properties and distribution was investigated. Experimental results showed the different biofilm thicknesses produced substantially different profiles of EPS composition and distribution. Soluble proteins were more dominant than soluble polysaccharides in the inner aerobic layer of the biofilms; in contrast, bound proteins were greater than bound polysaccharides in the outer anoxic or anaerobic layer of the biofilms. The biofilm-EPS matrix consisted mainly of bound EPS. Bound EPS exhibited a hump-shaped profile with the highest content occurring in an intermediate region in the thin MAB and relatively more uniformly in the one half of the biofilm close to the membrane side and then declined towards the biofilm-liquid interface in the thick MAB. The profiles of soluble EPS presented a similar declining trend from the membrane towards the outer region in both thin and thick MABs. The study suggested that not only EPS composition but also EPS distribution and properties (solubility and bindability) played a crucial role in controlling the cohesiveness and maintaining the structural stability and stratification of the MABs. PMID- 18403038 TI - Stress regulated expression of the GUS-marker gene (uidA) under the control of plant calmodulin and viral 35S promoters in a model fruit tree rootstock: Prunus incisa x serrula. AB - The fact that calmodulin genes (CaM) are tightly associated with the Ca(2+) regulatory pathway, as well as their putative role in plant defence against pathogens, indicate a potential use of alternative plant promoters to express genes of interest in specific tissues or developmental stages. To study the expression level of the apple CaM promoter, 981 bp sequences upstream were fused to the uidA gene, introduced into cherry and compared with a 35S-GUS construct. Transgene copy number and transgenic expression levels were analysed using Southern blot, Western blot and RT-PCR techniques. Transcription levels were assessed by GUS fluorometry, histochemistry and real-time PCR techniques in leaves of plantlets grown in vitro under various abiotic stresses like low- and high-temperature, salicylic acid and wounding, harvested after 0, 0.5, 1, 2, 4, 10, 24 and 72 h. Histochemical analyses showed staining only in veins and petioles of CaM-GUS lines, while in 35S-GUS plants staining extended to the entire leaf. Furthermore, real-time qPCR data indicate that both promoters are differently regulated by various stresses. Obtained results suggest that the selected apple CaM promoter responsible for the expression of a gene in vascular tissues may offer interesting perspectives for plant defense programs. PMID- 18403039 TI - Functional reconstitution of G-protein-coupled receptor-mediated adenylyl cyclase activation by a baculoviral co-display system. AB - Recently, evidence has accumulated in support of the heterologous expression of functional membrane proteins and their complexes on extracellular baculovirus particles (budded virus, BV). In this study, we attempted to apply this BV display system to detect G-protein-coupled receptor (GPCR) signaling. We infected Sf9 cells with a combination of four recombinant baculoviruses individually encoding the dopamine D1 receptor (DR-D1), G-protein alpha-subunit (Galpha(s)), G protein beta(1)gamma(2) subunit dimer (Gbeta(1)gamma(2)), and adenylyl cyclase type VI (ACVI). The recovered BV fraction produced cAMP in response to the stimulation with dopamine. Co-expression of all three G-protein subunits in addition to receptor and ACVI led to a maximal response. BV co-expressing DR-D1, Galpha(s), Gbeta(1)gamma(2), and ACVI also responded to dopamine agonists and an antagonist. Furthermore, BV expressing two other Galpha(s)-coupled receptors together with Galpha(s), Gbeta(1)gamma(2), and ACVI also produced cAMP in response to their specific ligands. These results indicate the functional coupling of receptor, Galpha(s) and ACVI is reconstituted on BV. Since BV is essentially free of endogenous GPCRs, this BV co-display system should prove highly useful for the development of functional assay systems for GPCRs. PMID- 18403040 TI - Evolutionary relationships of virus species belonging to a distinct lineage within the Ampelovirus genus. AB - A study of the evolutionary relationships of GLRaV-4,-5,-6 and -9, and two new Ampelovirus isolates (GLRaV-Pr and -De) related to grapevine leafroll disease was conducted based on molecular variability, positive selection analysis and maximum likelihood phylogenetic reconstructions. Sequences corresponding to the N terminal HSP70h and full CP encoding genes were determined for these viruses and datasets including homologous genomic regions from different members of the Closteroviridae were analyzed. GLRaV-Pr and -De were further characterised as distinct from the other closely related species after determination of a large genomic region (4319-4358 nts). ML phylogenetic topologies for both genes established the closer phylogenetic relationships of GLRaV-4,-5,-6,-9,-Pr and -De in regard to the other ampeloviruses, revealing very low inter-species evolutionary distances for this multitudinous lineage. The HSP70h segment phylogeny and bootstrap analysis enabled the identification of species within this lineage and provides a useful taxonomic tool for the rapid demarcation of these viruses. Estimations of d(N)/d(S) using the CP and HSP70h datasets revealed that, within the Closteroviridae, these viruses are subjected to the strongest constraints against amino acid substitutions. These estimations demonstrated a distinct evolutionary trait for this lineage probably related to its particular ecological niche that involves successful adaptation to the host, transmission through vegetative propagation and lack of vectors with high transmission efficiency. PMID- 18403041 TI - Functional mapping of the porcine reproductive and respiratory syndrome virus capsid protein nuclear localization signal and its pathogenic association. AB - PRRSV (porcine reproductive and respiratory syndrome virus) nucleocapsid (N) protein is the most abundant structural protein of the virus. During infection, the N protein is specifically localized to the nucleus and nucleolus in addition to its normal cytoplasmic distribution. Previously, a nuclear localization signal (NLS, 41-PGKK(N/S)KKKN)-null mutant virus (41-PGGGNKKKN) showed reduced viremia and increased production of neutralizing antibodies in infected pigs. However, the mutagenized NLS underwent strong selection pressure in the pig that resulted in partial or complete reversion and reacquisition of NLS function, and thus the biological effect of the NLS-null mutation needed further investigation. In the present study, a total of 9 "reversion resistant" mutants were generated by amino acid deletions and substitutions using an infectious cDNA clone. Two mutant clones (PG--SKKKS and PG--S-KKS) that produced progeny viruses were genetically stable for at least 20 passages in cell culture. Infection of pigs with those mutants induced neutralizing antibodies to higher titers than with wild-type virus. Both mutant viruses induced viremia of lower titer and of shorter duration than wild-type virus. RT-PCR from tonsils showed that both mutants persisted at a reduced level. Virus transmission to contact pigs was also lower in the mutant virus infected groups. No reversion to functional NLS was detected in either mutant from any pig. These data demonstrate that N protein nuclear localization is indeed associated with viral pathogenesis and host response to PRRS. PMID- 18403042 TI - Diagnosis of solitary lung nodules using the local form of Ripley's K function applied to three-dimensional CT data. AB - This paper analyzes the application of Ripley's K function to characterize lung nodules as malignant or benign in computerized tomography images. The proposed characterization method is based on a selection of measures from Ripley's K function to discriminate between benign and malignant nodules, using stepwise discriminant analysis. Based on the selected measures, a linear discriminant analysis procedure is performed once again in order to predict the classification of each nodule. To evaluate the ability of these features to discriminate the nodules, a set of tests was carried out using a sample of 39 pulmonary nodules, 29 benign and 10 malignant. A leave-one-out procedure was used to provide a less biased estimate of the linear discriminator's performance. The best setting of the analyzed function in the tested sample presented 70.0% of sensitivity but with 100.0% of specificity and 92.3% of accuracy. Thus, preliminary results of this approach are very promising regarding its contribution to the diagnosis of pulmonary nodules, but it still needs to be tested with larger series and associated to other quantitative imaging methods in order to improve global performance. PMID- 18403043 TI - Recent advances in the synthesis of aliphatic polyesters by ring-opening polymerization. AB - Advanced drug delivery systems rely on the availability of biocompatible materials. Moreover, biodegradability is highly desirable in the design of those systems. Consequently, aliphatic polyesters appear as a class of promising materials since they combine both properties. Nevertheless, their use in practical biomedical systems relies on clinical approval which not only depends on the material itself but also on its reproducible synthesis with the absence of residual toxics. The first sections of this review aim at reporting on the evolution of the initiators/catalytic systems and of the synthesis conditions (particularly the use of supercritical CO(2) as polymerization medium) in order to produce aliphatic polyesters with controlled macromolecular parameters by still "greener" ways. In addition, the further development of delivery systems also depends on the synthesis of materials exhibiting novel properties, such as amphiphilicity or pH-sensitivity that are emerging from the active research in macromolecular engineering. Functionalizing aliphatic polyesters is quite tedious due to their sensitivity towards hydrolytic degradation. The last section of this review is discussing several strategies to obtain functional (co)polyesters of various architectures providing them with novel properties. PMID- 18403044 TI - Advances in the synthesis of amphiphilic block copolymers via RAFT polymerization: stimuli-responsive drug and gene delivery. AB - Controlled/'living' radical polymerization methods, including the versatile reversible addition-fragmentation chain transfer (RAFT) polymerization process, are rapidly moving to the forefront in construction of drug and gene delivery vehicles. The RAFT technique allows an unprecedented latitude in the synthesis of water soluble or amphiphilic architectures with precise dimensions and appropriate functionality for attachment and targeted delivery of diagnostic and therapeutic agents. This review focuses on the chemistry of the RAFT process and its potential for preparing well-defined block copolymers and conjugates capable of stimuli-responsive assembly and release of bioactive agents in the physiological environment. Recent examples of block copolymers with designed structures and segmental compositions responsive to changes in pH or temperature are reviewed and hurdles facing further development of these novel systems are discussed. PMID- 18403045 TI - Predictors of talc pleurodesis outcome in patients with malignant pleural effusions. AB - OBJECTIVE: Chemical pleurodesis is an accepted palliative therapy for patients with recurrent, symptomatic, malignant pleural effusions (MPE). The purpose of the study was to determine the factors that have an effect on successful pleurodesis for MPE. PATIENTS AND INTERVENTIONS: Eighty-four consecutive patients with biopsy-proven malignant pleural disease and recurrent, symptomatic MPE were eligible to participate in this study. Five grams of talc mixed in 150ml of normal saline were administered via tube thoracostomy or small-bore catheters after complete drainage of the pleural effusion. RESULTS: Seven patients did not return for their 30-day follow-up visit and were excluded from further analysis. Successful pleurodesis was achieved in 63 of 77 eligible patients (81.8%) with MPE. In the univariate analysis, female gender, Karnofsky performance status, pleural fluid pH, cholesterol, and adenosine deaminase level showed a significant association with the probability of success. Multivariate logistic regression analysis showed that pleural fluid pH and ADA levels were independent predictors of talc pleurodesis outcome. CONCLUSION: Our results show that pleurodesis using talc as the sclerosing agent is a simple and acceptable procedure with high efficacy for controlling MPE, especially when used in appropriate patients. PMID- 18403046 TI - LAP2zeta binds BAF and suppresses LAP2beta-mediated transcriptional repression. AB - Proteins of the nuclear envelope have been implicated as participating in gene silencing. BAF, a DNA- and LEM domain-binding protein, has been suggested to link chromatin to the nuclear envelope. We have previously shown that LAP2beta, a LEM domain inner nuclear membrane protein, represses transcription through binding to HDAC3 and induction of histone H4 deacetylation. We now show that LAP2zeta, the smallest LAP2 family member, is also involved in regulation of transcription. We show that similar to other LEM-domain proteins LAP2zeta interacts with BAF. LAP2zeta-YFP and BAF co-localize in the cytoplasm, and overexpression of LAP2zeta leads to reduction of nucleoplasmic BAF. Mutations in the LAP2zeta-YFP LEM domain decrease its interaction with BAF retaining the nucleo-cytoplasmic distribution of BAF. Co-expression of LAP2beta and LAP2zeta results in inhibition of LAP2beta induced gene silencing while overexpression of LAP2zeta alone leads to a small increase in transcriptional activity of various transcription factors. Our results suggest that LAP2zeta is a transcriptional regulator acting predominantly to inhibit LAP2beta-mediated repression. LAP2zeta may function by decreasing availability of BAF. These findings could have implications in the study of nuclear lamina-associated diseases and BAF-dependent retroviral integration. PMID- 18403047 TI - Prevalence of latent tuberculosis infection in German radiologists. AB - Healthcare workers have an increased risk of tuberculosis infection compared with the general population. There have been few attempts to quantify the prevalence of latent tuberculosis infection amongst German healthcare workers, due to inadequacy of the current tuberculin skin test (TST). Therefore, it was our aim to investigate the prevalence of latent tuberculosis in this cohort using a tuberculosis-specific ELISpot (T-SPOT.TB) test and to compare the performance of this test to that of the TST. Ninety-five healthy participants working in departments of radiology were examined by ELISpot, lymphocyte transformation test and TST. For cellular in-vitro tests, tuberculosis-specific peptides and purified protein derivate (PPD) were used as antigens. These tests were combined with a questionnaire on prior tuberculosis exposure. Out of 95 healthcare workers, only one (1%) was defined as positive by T-SPOT.TB, 92 (97%) by PPD-ELISpot, 78 (82%) by PPD-lymphocyte transformation test and 32 (34%) by TST. Multivariate analysis showed that the TST was significantly affected (P<0.0001 and P=0.001, respectively) by foreign birth and prior skin testing. The T-SPOT.TB test results were independent of foreign birth, prior skin testing and prior vaccination against tuberculosis. In contrast to the TST, T-SPOT.TB appears to be an accurate and useful tool to track tuberculosis infection in this at-risk group. With only one of 95 participants having acquired latent tuberculosis, these preliminary results argue for a low incidence of latent tuberculosis in German radiologists. PMID- 18403048 TI - Comparative distribution of the mRNAs encoding urotensin I and urotensin II in zebrafish. AB - The neural neurosecretory system of fishes produces two biologically active neuropeptides, i.e. the corticotropin-releasing hormone paralog urotensin I (UI) and the somatostatin-related peptide urotensin II (UII). In zebrafish, we have recently characterized two UII variants termed UIIalpha and UIIbeta. In the present study, we have investigated the distribution of UI, UIIalpha and UIIbeta mRNAs in different organs by quantitative RT-PCR analysis and the cellular localization of the three mRNAs in the spinal cord by in situ hybridization (ISH) histochemistry. The data show that the UI gene is mainly expressed in the caudal portion of the spinal cord and, to a lesser extent, in the brain, while the UIIalpha and the UIIbeta genes are exclusively expressed throughout the spinal cord. Single-ISH labeling revealed that UI, UIIalpha and UIIbeta mRNAs occur in large cells, called Dahlgren cells, located in the ventral part of the caudal spinal cord. Double-ISH staining showed that UI, UIIalpha and UIIbeta mRNAs occur mainly in distinct cells, even though a few cells were found to co-express the UI and UII genes. The differential expression of UI, UIIalpha and UIIbeta genes may contribute to the adaptation of Dahlgren cell activity during development and/or in various physiological conditions. PMID- 18403049 TI - Oxytocin alleviates hepatic ischemia-reperfusion injury in rats. AB - Various mechanisms have been proposed for the pathogenesis of postischemic hepatic injury, including the generation of reactive oxygen metabolites. Oxytocin (OT) possesses antisecretory, antiulcer effects, facilitates wound healing and has anti-inflammatory properties. Hepatic ischemia-reperfusion (I/R)-injury was induced by inflow occlusion to median and left liver lobes ( approximately 70%) for 30 min of ischemia followed by 1h reperfusion in female Sprague-Dawley rats under anesthesia. I/R group (n=8) was administered intraperitoneally either OT (500 microg/kg) or saline at 24 and 12 h before I/R and immediately before reperfusion. Sham-operated group that underwent laparotomy without hepatic ischemia served as the control. Rats were decapitated at the end of reperfusion period. Hepatic samples were obtained for the measurement of myeloperoxidase (MPO) activity, malondialdehyde (MDA), glutathione (GSH) and collagen levels and histopathological analysis. Tumor necrosis factor-alfa (TNF-alpha) and transaminases (SGOT, SGPT) were assayed in serum samples. I/R injury caused significant increases in hepatic microscopic damage scores, MPO activity, collagen levels, transaminase, serum TNF-alpha levels. Oxytocin treatment significantly reversed the I/R-induced elevations in serum transaminase and TNF alpha levels and in hepatic MPO and collagen levels, and reduced the hepatic damage scores. OT treatment had tendency to abolish I/R-induced increase in MDA levels, while GSH levels were not altered. These results suggest that OT has a protective role in hepatic I/R injury and its protective effect in the liver appears to be dependent on its inhibitory effect on neutrophil infiltration. PMID- 18403050 TI - Human vasoactive hormone adrenomedullin and its binding protein rescue experimental animals from shock. AB - We recently discovered that vascular responsiveness to adrenomedullin (AM), a vasoactive hormone, decreases after hemorrhage, which is markedly improved by the addition of its binding protein AMBP-1. One obstacle hampering the development of AM/AMBP-1 as resuscitation agents in trauma victims is the potential immunogenicity of rat proteins in humans. Although less potent than rat AM, human AM has been shown to increase organ perfusion in rats. We therefore hypothesized that administration of human AM/AMBP-1 improves organ function and survival after severe blood loss in rats. To test this, male Sprague-Dawley rats were bled to and maintained at an MAP of 40 mmHg for 90 min. They were then resuscitated with an equal volume of shed blood in the form of Ringer's lactate (i.e., low-volume resuscitation) over 60 min. At 15 min after the beginning of resuscitation, human AM/AMBP-1 (12/40 or 48/160 microg/kg BW) were administered intravenously over 45 min. Various pathophysiological parameters were measured 4h after resuscitation. In additional groups of animals, a 12-day survival study was conducted. Our result showed that tissue injury as evidenced by increased levels of transaminases, lactate, and creatinine, was present at 4h after hemorrhage and resuscitation. Moreover, pro-inflammatory cytokines TNF-alpha and IL-6 were also significantly elevated. Administration of AM/AMBP-1 markedly attenuated tissue injury, reduced cytokine levels, and improved the survival rate from 29% (vehicle) to 62% (low-dose) or 70% (high-dose). However, neither human AM alone nor human AMBP-1 alone prevented the significant increase in ALT, AST, lactate and creatinine at 4h after the completion of hemorrhage and resuscitation. Moreover, the half-life of human AM and human AMBP-1 in rats was 35.8 min and 1.68 h, respectively. Thus, administration of human AM/AMBP-1 may be a useful approach for attenuating organ injury, and reducing mortality after hemorrhagic shock. PMID- 18403051 TI - Physiological significance of high- and low-affinity agonist binding to neuronal and recombinant AMPA receptors. AB - Radioligand binding studies have shown that AMPA receptors exist in two variants that differ about twenty-fold in their binding affinities, with brain receptors being mainly of the low-affinity type and recombinantly expressed receptors having almost exclusively high affinity. However, the physiological correlate of high- and low-affinity binding is not yet known. In this study we examined if physiological experiments similarly reveal evidence for two distinct receptor variants. We therefore measured equilibrium desensitization by glutamate and determined IC(50) values for neuronal receptors and for the homomeric receptors GluR1-4 expressed in HEK293 cells. Contrary to the prediction that these IC(50) values exhibit large differences commensurate with those of high- and low affinity binding, values for homomeric receptors (1-18 microM) were on an average not different from those of neuronal receptors (3-10 microM). Moreover, simulations with kinetic receptor models suggest that the IC(50) values for neuronal and recombinant receptors correspond to the binding affinity of the low affinity receptor variant. These findings indicate that the high-affinity binding measured in heterologous expression systems represents an immature receptor variant that does not contribute to the currents recorded from these cells, and that the functional low-affinity receptors are present in such small number that they are effectively masked in binding assays by the high-affinity receptors. Thus, in order to compare experimentally determined saturation binding profiles with those predicted by kinetic receptor models and with dose-response curves from physiological studies, it will be imperative to develop methods for isolating first the low-affinity receptors. PMID- 18403053 TI - Proteasome inhibition increases tau accumulation independent of phosphorylation. AB - An intrinsic link between proteasome and tau degradation in Alzheimer's disease (AD) has been suggested, however, the role of proteasome in the proteolysis of tau is still uncertain. Here, we investigated the influence of proteasome inhibition on the accumulation, phosphorylation, ubiquitination, solubility of tau and the memory retention in rats. We observed that lactacystin inhibited the proteasome activities and increased the level and insolubility of different tau species, including phosphorylated tau. The elevation of the phosphorylated tau was no longer present and the level of pS214 and pT231 tau was even lower than normal level after normalized to total tau. Inhibition of proteasome resulted in activation of cAMP-dependent protein kinase, glycogen synthase kinases-3beta and cyclin-dependent kinase-5, and inhibition of protein phosphatase-2A and c-Jun N terminal kinase (JNK). Proteasome inhibition did not affect the memory retention of the rats. We conclude that proteasome inhibition increases accumulation and insolubility of tau proteins independent of tau phosphorylation, and JNK inhibition may be partially responsible for the relatively decreased phosphorylation of tau in the rat brains. PMID- 18403052 TI - Co-localization of the amyloid precursor protein and Notch intracellular domains in nuclear transcription factories. AB - The beta-amyloid precursor protein (APP) plays a major role in Alzheimer's disease. The APP intracellular domain (AICD), together with Fe65 and Tip60, localizes to spherical nuclear AFT complexes, which may represent sites of transcription. Despite a lack of co-localization with several described nuclear compartments, we have identified a close apposition between AFT complexes and splicing speckles, Cajal bodies and PML bodies. Live imaging revealed that AFT complexes were highly mobile within nuclei and following pharmacological inhibition of transcription fused into larger assemblies. We have previously shown that AICD regulates the expression of its own precursor APP. In support of our earlier findings, transfection of APP promoter plasmids as substrates resulted in cytosolic AFT complex formation at labeled APP promoter plasmids. In addition, identification of chromosomal APP or KAI1 gene loci by fluorescence in situ hybridization showed their close association with nuclear AFT complexes. The transcriptional activator Notch intracellular domain (NICD) localized to the same nuclear spots as occupied by AFT complexes suggesting that these nuclear compartments correspond to transcription factories. Fe65 and Tip60 also co localized with APP in the neurites of primary neurons. Pre-assembled AFT complexes may serve to assist fast nuclear signaling upon endoproteolytic APP cleavage. PMID- 18403054 TI - PSEN1 polymorphisms alter the rate of cognitive decline in sporadic Alzheimer's disease patients. AB - Mutations in amyloid precursor protein (APP) and presenilin (PSEN) genes are known to cause familial early-onset Alzheimer's disease (AD), which account for around 5% of AD cases. Genetic associations for the remaining "sporadic" cases, other than the risks associated with the apolipoprotein (APOE) epsilon4 allele are currently not fully established. The aim of this study was to investigate whether single nucleotide polymorphisms (SNPs) in PSEN1 are associated with a modified risk for sporadic AD or a modified disease phenotype. Eight tag SNPs were identified using linkage disequilibrium (LD) data from the International HapMap project providing coverage of the entire PSEN1 gene. These SNPs were investigated for AD susceptibility in a case-control haplotype association study (N=714) and for genotype-specific effects on cognitive performance in AD patients (N=169) using non-linear mixed effects modelling. Replication of a mild associated-risk of an intronic PSEN1 polymorphism with AD was achieved (P=0.03). No other single SNPs or haplotypes were associated with AD risk. However, 3 SNPs were associated with an altered rate of cognitive decline underlining their role as genetic modifiers of disease. PMID- 18403055 TI - CSF biomarker levels in early and late onset Alzheimer's disease. AB - OBJECTIVE: To compare CSF levels of beta-amyloid 1-42 (Abeta(1-42)), total tau (tau) and tau phosphorylated at threonine 181 (ptau-181) between AD patients and controls according to age. METHODS: 248 AD patients (48% men) and 127 controls (51% men, 22 volunteers and 105 subjective complainers) underwent lumbar puncture. Both patients and controls were divided into a young (<65 years) and old (>or=65 years) group. RESULTS: All three biomarkers showed main effects of diagnosis (p<0.001). There was an interaction between diagnosis and age for all three biomarkers (p<0.05), as old controls had lower Abeta(1-42) and higher (p)tau than young controls (Abeta(1-42) 699+/-250 versus 866+/-191pg/ml, tau 408+/-245 versus 243+/-102pg/ml, ptau-181 60+/-28 versus 42+/-15pg/ml), but there was no difference according to age among AD patients (Abeta(1-42) 451+/-178 versus 425+/-146pg/ml, tau 741+/-460 versus 798+/-467pg/ml, ptau-181 91+/-42 versus 91+/-41pg/ml). CONCLUSION: We found that the older control group had lower Abeta(1-42) and higher (p)tau compared to the younger control group. This suggests that older individuals may have AD pathology, even in the absence of objective cognitive impairment. PMID- 18403056 TI - Ligand binding to nucleic acids and proteins: Does selectivity increase with strength? AB - The possible relation of strength and selectivity of ligand binding to biomacromolecules and its theoretical limitation is discussed and illustrated with some examples. It is shown that a linear correlation between selectivity and affinity may be expected on the basis of thermodynamic principles, which also imply that multivalency is as important for selectivity as for affinity enhancement. That strictly linear correlations are often not observed is, apart form statistical problems, mostly due to interactions which may remain constant only at some sites but can differ significantly at other sites, which, e.g., dominate the affinity. Nevertheless, some drugs exhibit in line with theory at the same time a peak affinity and selectivity, such as etonitazene with different opioid receptors. Double-stranded nucleic acids feature relative stable and uniform structures and therefore show relatively good correlations with simple polyamines as ligands and RNA or DNA model receptors. Metalloproteins possess strong binding centers with additional discrimination sites, and can exhibit linear correlations, at least with structurally related metalloproteinases and their inhibitors. PMID- 18403057 TI - QSAR study of antioxidant activity of wine polyphenols. AB - Quantitative structure activity relationships (QSAR) were obtained describing the antioxidant activity of the main pharmacologically active polyphenols of wine, using molecular properties and descriptors derived from the 2D and 3D representations of molecules. The best models for the prediction of the ability to scavenge the ABTS radical cation were obtained by polynomial regression analysis using zero-order connectivity index and molar refractivity. Statistically, significant models for lipid peroxidation inhibiting effects of flavonoids were obtained by polynomial and multiple regression using lipophilicity, Balaban index, Balaban-type index and 3D GETAWAY descriptor. The 3D descriptors possess the ability for discrimination of stereoisomers, like catechin and epicatechin. We demonstrated that a size and shape of molecules, as well as steric properties, play an important role in the antioxidant activity of polyphenols. PMID- 18403058 TI - Novel imidazo[1,2-a]naphthyridinic systems (part 1): synthesis, antiproliferative and DNA-intercalating activities. AB - Novel imidazo[1,2-a]naphthyridinic systems 6a-15a and 6b-15b were obtained from Friedlander's reaction in imidazo[1,2-a]pyridine series. Most of the compounds were evaluated for their antitumor activity in the NCIs in vitro human tumor cell line screening panel. Among them, pentacyclic derivatives 13b and 14a exhibited in vitro activity comparable to anticancer agent such as amsacrine. Their mechanism of cytotoxicity action was unrelated to poisoning or inhibiting abilities against topo1. On the contrary, we highlighted a direct intercalation of the drugs into DNA by electrophoresis on agarose gel. PMID- 18403059 TI - 2-[2-Substituted-3-(3,4-dichlorobenzylamino)propylamino]-1H-quinolin-4-ones as Staphylococcus aureus methionyl-tRNA synthetase inhibitors. AB - New analogues of 2-[2-substituted-3-(3,4-dichlorobenzylamino)propylamino]quinolin 4-ones, 26a, 26b, 31a-e, 34, 35, 38 and 40, have been synthesized and evaluated against Staphylococcus aureus methionyl-tRNA synthetase. All of the synthesized compounds were less active than the reference compound 2. The compounds were also screened against various strains of S. aureus and Enterococci for their antibacterial activities. Among the compounds, 26b, 31c and 31e displayed significant inhibitory properties against various strains of Enterococci compared to compound 2. PMID- 18403060 TI - Newer GABA derivatives for the treatment of epilepsy including febrile seizures: a bioisosteric approach. AB - The present study aims at design and synthesis of newer gamma-aminobutyric acid (GABA) derivatives with the combination of thiosemicarbazone and GABA pharmacophores in order to develop newer anticonvulsants. The reported compounds were designed as bioisosteric analogues of GABA semicarbazones. The structures of the synthesized compounds were confirmed by the use of their spectral data besides elemental analysis. Initial anticonvulsant screening was performed using intraperitoneal (i.p.) maximal electroshock-induced seizure (MES), subcutaneous pentylenetetrazole (scPTZ), subcutaneous strychnine (scSTY), and subcutaneous picrotoxin (scPIC)-induced seizure threshold tests. A model involving 22-day old rat pups was also employed to further screen the effects of the test compounds against hyperthermia-induced febrile seizures. Only compounds 1 and 11 were found to be active in the MES test. Most of the compounds were found to be effective in the scPIC and febrile seizure models and very few compounds showed protection in scPTZ and scSTY models. This is the first report on these new GABA derivatives effective in the treatment of febrile seizures. PMID- 18403061 TI - [B lymphocytes in Sjogren's syndrome]. AB - INTRODUCTION: Sjogren's syndrome (SS) is an autoimmune epithelitis hallmarked by a disruption of epithelial cells, the subsequent lymphocytic infiltration of lachrymal and salivary glands (SGs), and their ensuing dryness. One may posit that SS is triggered by viruses, and/or modulated by sex steroid hormones, and there is indeed a consensus that its aetiology is multifactorial, with genetic factors interacting with environmental agents. CURRENT KNOWLEDGE AND KEY POINTS: T-cells have long occupied central stage of the debate on the type of lymphocytes involved in the pathogenesis of SS. The relevance of B cells has, however, been emphasized over the past five years and new insights into their functions revealed. Furthermore, increased levels of the B-cell activating factor (BAFF) may be responsible for quantitative and qualitative anomalies of B-cells found in SS such as emergence of self reactive B-cells. This review reports compelling evidence that B-cells are involved in the pathophysiology of SS. PROSPECTS: Since SS may thus be conceived as a model for B-cell-induced autoimmunity, it is no surprise that B-cell ablative-treatment has proven to be relatively effective in SS. PMID- 18403062 TI - [Risk of venous thromboembolism in the course of acute medical illness. Part 1: Basic research, clinical models, descriptive and analytic epidemiology]. AB - PURPOSE: Some acute medical illnesses are associated with an increased risk of venous thromboembolism. This risk is difficult to assess in medical patients on account of the multiplicity of their pathologies, leading to potential interactions between numerous risk factors. CURRENT KNOWLEDGE AND KEY POINTS: A rational analysis of the risk of venous thrombosis in acute medical setting may be derived from pathogenic models and be based on elementary risk factors for venous thromboembolism classified as transient, permanent, major and intermediate factors. The systematic study of pathophysiological mechanisms in process, in each medical patient could guide the physician attitude toward still non evaluated situations. FUTURE PROSPECTS AND PROJECTS: A synthesis of the last epidemiological and pathophysiological data is developed in the first part of this literature review, whereas the risk linked to specific situations in ambulatory, hospital or internal medicine settings will be described in the second part. PMID- 18403063 TI - [What's new in autoinflammatory diseases?]. AB - PURPOSE: The concept of auto-inflammation was initially coined to define the group of mendelian disorders characterized by recurrent inflammatory symptoms. The core of this group mainly consists of hereditary recurrent fevers, which has been lately enlarged to other inflammatory mendelian disorders as well as to some sporadic diseases with a genetic component relevant to innate immunity. CURRENT KNOWLEDGE AND KEY POINTS: Cryopyrin, the product of the CIAS1/PYPPAF1/NALP3/NLRP gene, whose mutations underline some mendelian syndromes (Muckle-Wells and chronic infantile neurological cutaneous and articular (CINCA), familial cold urticaria) can now be considered as a major factor of the regulation of interleukin-1 production within the multiprotein complex called inflammasome. This discovery has lit up our view of innate immunity. FUTURE PROSPECTS AND PROJECTS: The contribution of the innate immunity mechanisms in inflammatory disorders have led to a new look to the current nosology of this vast group of diseases and to suggest a classification with two poles. The first would be defined by the predominance of auto-inflammation, whereas in the second one auto immunity predominates. PMID- 18403064 TI - [Acquired factor XI inhibitor and chronic lymphocytic leukemia]. AB - Autoimmune phenomena, most frequently autoimmune hemolytic anemia, is a well known complication of lymphoproliferative diseases. We report a very rare association of a chronic lymphocytic leukemia with an acquired factor XI inhibitor. A 87-year-old man presented with auto-immune hemolytic anemia. He had untreated chronic lymphocytic leukemia for the past three years and renal insufficiency. Before surgical procedure for arteriovenous fistula, we discovered a very prolonged activated partial thromboplastin time (APTT), and an acquired factor XI inhibitor was detected. The patient was successfully treated with immunosuppressive therapy. Among patients with lymphoproliferative disorders the discovery of a prolonged APTT implies to search for rare autoimmune phenomena like acquired coagulation factor inhibitors. PMID- 18403065 TI - [Fever of unknown origin: diagnostic strategies and tactical approaches]. AB - PURPOSE: Diagnosis of fever of unknown origin (FUO) is a major challenge for internists, as emphasized by the high rate of diagnostic failure, despite the fast-moving progress in medical technology. Numerous clues are available in clinical and standard biological data; a better use of the available tests is warranted. KEY POINTS: Improvement in diagnostic accuracy might be expected by developing strategies targeted toward a more systematic search of diagnosis clues. Intuition and the hypothetic-deductive method that are the most common clinical strategies are the most perfectible. It implies to enjoy the fun of clinical examination, to have a large experience in bedside training, to be confident in his/her own semiological skills, to refer frequently to heuristics, and to use carefully Occam's razor principle. Laboratory tests might be revisited; immunological and serological tests are of little value; standard biological tests provide many insufficiently exploited clues. Imaging procedures depend on objectives: whole body CT scan should be performed early within the first days of hospitalisation, preceded by standard chest radiograph and abdomen ultrasonography; followed by either indium-111 or technetium-99m, labelled leukocytes if deep abscesses are suspected or 18-FDG PET scan in the case of suspected inflammatory disease involving tissues, lymph nodes or arteries. Early identification of the best tissue to be the site of biopsy is one of the most decisive procedures. CONCLUSION: Strategies and tactical approaches for the diagnosis of FUO might be driven by the search of significant clues. Self clinical experience driven by a wide bedside training is of major concern. Standard laboratory tests might be better used and the choice of imaging depends on objectives. Identification of the most appropriate tissue to be sampled for histological examination is one of the most beneficial step. PMID- 18403066 TI - [Is rituximab a promising treatment for Sjogren's syndrome?]. PMID- 18403067 TI - [National ranking examination four years after its implementation: thoughts]. PMID- 18403068 TI - Protection of chickens from fowl cholera by vaccination with recombinant adhesive protein of Pasteurella multocida. AB - The recombinant adhesive protein (rCp39) of Pasteurella multocida strain P-1059 (serovar A:3) was prepared and purified with a hybrid condition of affinity chromatography. The rCp39 was highly protective for chickens from fowl cholera by challenge-exposure with parental strain P-1059 or heterologous strain X-73 (serovar A:1) compared to various kind of vaccines. Sixteen groups of ten chickens each were subcutaneously inoculated twice with 100, 200 or 400 microg proteins of rCp39, native Cp39, native outer membrane protein H (OmpH) or recombinant OmpH, or 100 microg proteins of crude capsular extract (CCE) of strains P-1059 or X-73 at 2 weeks interval. Five chickens of each group were challenge-exposed with each strain 2 weeks after the second inoculation. As the results, 60-100% protections were demonstrated in the chickens against both strains. Fisher's exact test indicated no significant differences (P<0.05) in vaccine types and dosages. ELISA and Western blot analysis indicated that the chicken anti-rCp39 sera reacted to whole-cell lysate of parental or heterologous strains. In conclusion, rCp39 is a cross-protective recombinant adhesive antigen of P. multocida capsular serogroup A strains. Moreover, a hybrid condition of affinity chromatography was successfully demonstrated and protected the immunogenicity of recombinant protein. PMID- 18403069 TI - Effectiveness of hexavalent vaccines against invasive Haemophilus influenzae type b disease: Germany's experience after 5 years of licensure. AB - Vaccine effectiveness (VE) was determined with a case-cohort approach using Cox regression. Cases with confirmed systemic Hib infections in children born from 1 August 2000 to 31 December 2004 were ascertained through two independent nationwide active surveillance systems. A representative cohort of 1303 children born in the same time frame was randomly sampled in a nationwide immunisation survey. Thirty cases were eligible for VE calculation; 19 were unvaccinated and 11 vaccinated with hexavalent vaccines. VE was 68.4% (95% CI: 19.0-87.6) for incomplete primary series and 90.4% (95% CI: 70.6-96.8) for the full primary series. For full immunisation VE was 100.0% (95% CI: 52.7-100.0). Hexavalent vaccines show a high effectiveness against invasive Hib disease in Germany. PMID- 18403070 TI - Immunization with an engineered mutant trans-sialidase highly protects mice from experimental Trypanosoma cruzi infection: a vaccine candidate. AB - Chagas' disease is a major tropical disease for which a cure for chronic phase does not exist yet. Trypanosoma cruzi trans-sialidase (TS) seems to be involved in relevant processes such as infectivity, host survival and, very importantly, disease pathogenesis. In this study, we show that mice vaccinated with an engineered enzymatically deficient mutant TS containing the catalytic domain without the immunodominant SAPA (Shed Acute Phase Antigen) repeats, were highly protected against T. cruzi infection. Adult male BALB/c mice were immunized with mutant protein, purified from Pichia pastoris yeast, using three inoculations in Freund's adjuvant. All immunized mice were protected against challenge with a lethal dose of T. cruzi trypomastigotes. The protected immunized mice developed no clinical or tissue evidence of infection throughout the study. In contrast, 60 90% mortality and 100% occurrence of myocardial lesions were observed in the non immunized counterparts. Titers of circulating antibody against TS did not correlate with protection, while anti-SAPA antibodies were coincident with disease severity. Further studies indicated that a single inoculation of mutant recombinant protein in Freund's complete adjuvant was not associated with blood or organic alterations, per se. Mutant TS vaccination seems to be a promising tool for immune intervention strategies in Chagas' disease, aimed at preventing T. cruzi-related heart tissue damage. PMID- 18403071 TI - Safety evaluation of pulmonary influenza vaccination in healthy and "asthmatic" mice. AB - The present study reports animal immuno-toxicological data of pulmonary vaccination against inactivated seasonal influenza. Its aims were (i) to monitor the temporal kinetics of lung inflammation in normal mice over a period of 2 weeks following pulmonary vaccination in order to assess the risk of chronic lung inflammation, (ii) to evaluate the impact of pulmonary vaccination on the asthmatic phenotype in an established allergen-sensitized murine model of asthma. Both sets of experiments were performed using high doses of split influenza virus vaccine. In the first part of this study, we showed that pulmonary influenza vaccination induced a slight local inflammatory response which was limited in duration since it was no longer observed at 2 weeks post-vaccination. At this time point, it has previously been shown that the immunogenic efficacy was maintained. In the second part, we demonstrated that pulmonary influenza vaccination did not significantly exacerbate the cardinal features of asthma, i.e., allergen-specific IgE formation, the development of airway hyperreactivity (AHR) and eosinophilic airway inflammation. Our data therefore suggest that the overall immuno-toxicological profile of pulmonary vaccination against seasonal influenza was acceptable, even in an animal model of pulmonary hypersensitivity. PMID- 18403072 TI - Global implications of evidence 'biased' practice: management of the third stage of labour. PMID- 18403073 TI - Stature loss and recovery following a period of loading: effect of time of day and presence or absence of low back pain. AB - BACKGROUND: Stature reductions in asymptomatic individuals, caused by a set load, are lower later in the day when stature is in the trough of diurnal variation; hence most stature reduction investigations are conducted in the morning. Recent evidence suggests that it is not the reductions in stature, but the recovery of stature, that is of greatest importance. The aim of this investigation was to establish whether stature recovery is also affected by time of day and to determine if any differences exist between a chronic low back pain and asymptomatic group. METHODS: Eleven chronic low back pain participants (age=32.8 SD 7.9 yrs, mass=74.4 SD 14.2 kg and height=1.73 SD 0.07 m) and 11 asymptomatic participants (age=31.0 SD 6.3 yrs, body mass=72.6 SD 11.5 kg and height=1.76 SD 0.09 m) underwent two 20 min loaded walking tasks (10% body mass), one in the morning (09:00) and one in the afternoon (14:00), followed by a 20 min unloaded recovery period. Measurements of stature were obtained throughout. FINDINGS: The asymptomatic group experienced significantly less stature reduction (P=0.05; ES=1.1) and greater stature recovery (P=0.02; ES=0.9) in the afternoon compared to the morning. The chronic low back pain group experienced a similar pattern to the asymptomatic group, however no significant difference between sessions for changes in stature was evident P=0.07. INTERPRETATION: Further investigations of stature recovery should be restricted to the morning when comparing individuals with and without chronic low back pain, as time of day appeared to have effect on stature recovery, particularly in the asymptomatic group. Time dependent differences in stature change between these two populations warrants further investigation. PMID- 18403074 TI - Changes in cell-type composition in digestive gland of slugs and its influence in biomarkers following transplantation between a relatively unpolluted and a chronically metal-polluted site. AB - Changes in cell-type composition (CCTC) is a general phenomenon that takes place in the digestive gland epithelium of stressed molluscs. The aim of the present work was to determine whether CCTC is a reversible process in the digestive gland of sentinel slugs chronically exposed to metal pollution and how CCTC affects metal accumulation parameters and different cell and tissue biomarkers of exposure and effect. Slugs (Arion ater) from an abandoned zinc mine were transferred to a relatively unpolluted site and the other way around for 3, 10 and 28 d. The volume density of black silver deposits (Vv(BSD)) after autometallography, and metallothionein (MT) levels were used as biomarkers of exposure to metals and CCTC and lysosomal responses were selected as effect biomarkers. Results indicated that slugs were sensitive to recent metal pollution; however, slugs chronically exposed to metals presented some characteristic features and were less responsive to pollution cessation without signs of CCTC reversal. PMID- 18403075 TI - New metallothionein assay in Scrobicularia plana: heating effect and correlation with other biomarkers. AB - Metallothionein (MT) and other biomarker levels were measured in Scrobicularia plana clams to assess pollution of the Guadalquivir Estuary possibly affected by metals released from Aznalcollar pyrite mine in 1998. After optimizing reagent concentrations for monobromobimane derivatization, MT levels were quantified by reversed-phase high-performance liquid chromatography coupled to fluorescence detection (RP-HPLC-FD) in heated or unheated digestive gland extracts and compared to those obtained by differential pulse polarography (DPP). MT content assayed by RP-HPLC-FD in unheated samples was higher than that obtained by DPP and correlated better with metals and anti-oxidant activities. MT assay by RP HPLC-FD in unheated extracts would be preferable for assessing metal pollution, due to its greater sensitivity and specificity. In addition to MT induction, glyoxalase II inhibition was well correlated with metal contents. Our results suggest that metals at the estuary do not originate from Aznalcollar spill, but from those carried along by the river and deposited at its concave bank. PMID- 18403076 TI - The research, policy and practice interface: reflections on using applied social research to promote equity in health in Malawi. AB - The case for research to promote equity in health in resource poor contexts such as Malawi is compelling. In Malawi, nearly half of all the people with tuberculosis cannot afford to access free tuberculosis services. In this scenario, there is a clear need to understand the multiple barriers poor women and men face in accessing services and pilot interventions to address these in a way that engages policy makers, practitioners and communities. This paper provides a critical reflection on our experience as applied social researchers working at the REACH (Research for Equity and Community Health) Trust in Malawi. Our work largely uses qualitative research methodologies as a tool for applied social research to explore the equity dimensions of health services in the country. We argue that a key strength of qualitative research methods and analysis is the ability to bring the perceptions and experiences of marginalised groups to policy makers and practitioners. The focus of this paper is two-fold. The first focus lies in synthesising the opportunities and challenges we have encountered in promoting the use of applied social research, and in particular qualitative research methods, on TB and HIV in Malawi. The second focus is on documenting and reflecting on our experiences of using applied social research to promote gender equity in TB/HIV policy and practice in Malawi. In this paper, we reflect on the strategic frameworks we have used in the Malawian context to try and bring the voices of poor women and men to policy makers and practitioners and hence intensify the research to policy and practice interface. PMID- 18403078 TI - Tacit and encoded knowledge in the use of standardised outcome measures in multidisciplinary team decision making: a case study of in-patient neurorehabilitation. AB - This paper explores how multidisciplinary teams (MDTs) balance encoded knowledge, in the form of standardised outcome measurement, with tacit knowledge, in the form of intuitive judgement, clinical experience and expertise, in the process of clinical decision making. The paper is based on findings from a qualitative case study of a multidisciplinary in-patient neurorehabilitation team in one UK NHS trust who routinely collected standardised outcome measures. Data were collected using non-participant observation of 16 MDT meetings and semi-structured interviews with 11 practitioners representing different professional groups. Our analysis suggests that clinicians drew on tacit knowledge to supplement, adjust or dismiss 'the scores' in making judgements about a patients' likely progress in rehabilitation, their change (or lack of) during therapy and their need for support on discharge. In many cases, the scores accorded with clinicians' tacit knowledge of the patient, and were used to reinforce this opinion, rather than determine it. In other cases, the scores, in particular the Barthel Index, provided a partial picture of the patient and in these circumstances, clinicians employed tacit knowledge to fill in the gaps. In some cases, the scores and tacit knowledge diverged and clinicians preferred to rely on their clinical experience and intuition and adjusted or downplayed the accuracy of the scores. We conclude that there are limits to the advantages of quantifying and standardising assessments of health within routine clinical practice and that standardised outcome measures can support, rather than determine clinical judgement. Tacit knowledge is essential to produce and interpret this form of encoded knowledge and to balance its significance against other information about the patient in making decisions about patient care. PMID- 18403077 TI - Promoting the social value of research in Kenya: examining the practical aspects of collaborative partnerships using an ethical framework. AB - The ethics of research continue to attract considerable debate, particularly when that research is sponsored by partners from the North and carried out in the South. Ethical research should contribute to social value in the country where research is being carried out, but there is significant debate around how this might be achieved and who is responsible. The literature suggests that researchers might employ two inter-related strategies to maximise social value: collaborative partnerships with policy makers and communities from the outset of research, and dissemination of research results to participants, policy makers and implementers once the research is over. These areas have received relatively little empirical attention. In this study, we carried out 40 in-depth interviews to explore the role of collaborative partnerships in health research priority setting, and the way in which research findings are disseminated to aid policy making and implementation in Kenya. Interviewees included policy makers, researchers, policy implementers and representatives of organisations funding health reforms in Kenya. Two policy issues were drawn upon as tracers wherever possible: (1) the introduction of Artemesinin-based Combination Therapies (ACTs), an anti-malarial treatment policy; and (2) Haemophilus influenzae (Hib) vaccine for the prevention of pneumococcal diseases among children. The findings point to significant gaps in the 'research to policy to practice' pathway, particularly for national research institutions with a focus on clinical/biomedical research. These gaps reflect poorly effective partnerships among stakeholders and limit the potential social value of much research. While more investment is needed to establish strong structures for promoting and directing collaboration and partnership, how to target this investment is not entirely clear, especially in the context of the considerable power of the global health agenda and the research financing tied to it. PMID- 18403079 TI - Ethical dilemmas of social science research on AIDS and orphanhood in Western Kenya. AB - This paper is based on the experiences drawn from a long-term social science research programme on the impact of the AIDS pandemic on orphanhood in western Kenya. It discusses the ethical dilemma of maintaining a delicate balance between research ethics, the expectations of the study population and negotiating the community's vested interests in a health related research project in a low-income society. I argue that informed consent and the intended benefits of the study to the participants continue to be major challenges facing the justification of social research with people affected by or living with AIDS in low-income societies. The paper underscores the importance of community feedback sessions as a way of enhancing chances of acceptability of research efforts and obtaining informed consent. It further shows how community feedback sessions contribute to local knowledge of the problem being studied, creating opportunities for advocacy. This discussion adds to the existing ethical debate on the wider contexts within which research on vulnerable people affected by AIDS is conducted by arguing that research practice is inseparable from epistemological concerns of knowledge production. I suggest that ethnographers should enhance efforts to innovatively design action research projects to serve the twin purposes of data collection and deal with ethical challenges that are experienced when doing long term research on vulnerable groups. PMID- 18403080 TI - Differential effects of aging on processes underlying task switching. AB - In this study, we used event-related brain potentials (ERPs) to examine the effects of aging on processes underlying task switching. The response time data revealed an age-related increase in mixing costs before controlling for general slowing and no effect of aging on switching costs. In the cue-locked epoch, the ERP data revealed little effect of age on the parietal P3 related to cue encoding, an age-related decrease in parietal activity related to cue retrieval, and an age-related increase in the amplitude of the parietal and frontal activity related to task set configuration and rule mapping. In the target-locked epoch, there was differential neural recruitment in younger and older adults in response to task mixing. These data are consistent with the idea that older adults may not fully implement task set before onset of the target stimulus. PMID- 18403081 TI - Efficacy of electroconvulsive therapy is associated with changing blood levels of homovanillic acid and brain-derived neurotrophic factor (BDNF) in refractory depressed patients: a pilot study. AB - Electroconvulsive therapy (ECT) is effective for patients with antidepressant medication-resistant depression. However, the mechanisms of ECT's effectiveness for treating depression are not fully understood. We therefore investigated ECT's effects on blood levels of brain-derived neurotrophic factor (BDNF), catecholamine metabolites, and nitric oxide (NO) in 18 treatment-refractory depressed patients. Serum BDNF levels increased significantly following ECT in responders to ECT (before ECT: 8.0+/-9.7 ng/mL; five weeks after start of ECT: 15.1+/-11.1 ng/mL), whereas BDNF levels in non-responders were unchanged (before ECT: 11.5+/-11.0 ng/mL; five weeks after start of ECT: 9.4+/-7.5 ng/mL). Furthermore, the plasma HVA levels, but not MHPG levels, were significantly reduced after ECT (before ECT: 8.5+/-1.9 ng/mL; five weeks after start of ECT: 5.8+/-2.2 ng/mL). This latter finding occurred in parallel with the improvement of depressive symptoms in all patients. These results suggest that the mechanisms underlying ECT's effect on refractory depression may be related to dopaminergic neurons and BDNF. PMID- 18403082 TI - Effect of N-acetylcysteine and/or deferoxamine on oxidative stress and hyperactivity in an animal model of mania. AB - Studies have consistently reported the participation of free radicals in Bipolar Disorder. Administration of d-amphetamine (d-AMPH) is a relevant animal model of mania and it increases oxidative stress in rat brain. Evidences indicate that the antioxidants N-acetylcysteine (NAC) and Deferoxamine (DFX) exert protective effects in the brain. The present study was designed to evaluate the effects of NAC, DFX or their combination on AMPH-induced hyperactivity. The protein oxidation levels were analyzed in prefrontal cortex and hippocampus. In the first animal model (reversal treatment), adult male Wistar rats received saline or d AMPH for 14 days, and from the 8th to the 14th day, they were treated with saline, NAC, DFX, or NAC plus DFX. In the second animal model (prevention treatment), rats were pretreated with saline or antioxidant regime, and from the 8th to the 14th day, they also received saline or d-AMPH. In the prefrontal cortex, the protein carbonyls were not affected by the treatment with antioxidants alone but it was increased by treatment with NAC plus DFX. At the same model, NAC plus DFX reversed the protein damage in the hippocampus, but NAC alone increased this damage. In the prevention treatment, it was observed that the protein damage in the prefrontal cortex was prevented by DFX or NAC plus DFX. In the hippocampus, the pretreatment with all antioxidant regime prevented protein damage induced by d-AMPH. At both treatments (reversal or prevention) the antioxidants did not present any effect against d-AMPH-induced hyperactivity. In conclusion, NAC or DFX and the combination of NAC plus DFX reverse and protect against d-AMPH-induced oxidative protein damage. Using these protocols we could not observe affects on locomotion, however this effect varies depending on the brain region and the treatment regime. PMID- 18403083 TI - Subchronic feeding study of herbicide-tolerant soybean DP-356O43-5 in Sprague Dawley rats. AB - Optimum GAT1 soybean is a genetically modified (GM) soybean containing event DP 356O43-5 (356043) that was produced by integration of the coding sequences of the GAT4601 and GM-HRA proteins. In planta expression of these proteins confers tolerance to glyphosate and sulfonylurea/imidazolinone herbicides, respectively. This paper reports the results from a subchronic rat feeding study conducted with 356043 soybeans. Dehulled/defatted toasted meal and toasted ground hulls were prepared from soybeans from untreated plants (356043), herbicide-treated plants (356043+Gly/SU), non-transgenic isoline control (091), and three commercial non transgenic reference varieties (93B86, 93B15, and 93M40). Individual diets conforming to standard certified rodent chow formulation (Purina Rodent LabDiet) 5002) were prepared with 20% meal (w/w) and 1.5% hulls (w/w). Diets were fed to young adult Sprague-Dawley rats (12/sex/group) for at least 93 days. Compared with rats fed the isoline control or conventional reference diets, no biologically-relevant, adverse effects were observed in rats fed diets containing 356043 or 356043+Gly/SU soybean with respect to body weight/gain, food consumption/efficiency, clinical signs, mortality, ophthalmology, neurobehavioral assessments (sensory response, grip strength, motor activity), clinical pathology (hematology, coagulation, serum chemistry, urinalysis), organ weights, and gross and microscopic pathology. The results from this study indicate that 356043 soybeans are as safe and nutritious as conventional non-GM soybeans. PMID- 18403084 TI - Inhibitive effects of enhanced lipid peroxidation on Ca(2+)-ATPase in lenses of hereditary cataract ICR/f rats. AB - Our previous studies have demonstrated that the instillation of eye drops containing disulfiram, a radical scavenger and nitric oxide synthase inhibitor, delays cataract development in ICR/f rats, and we have suggested that the production of nitric oxide (NO) and lipid peroxide (LPO) in the lens may relate to the delay in cataract development brought about by disulfiram. However, the involvement of NO and LPO in lenses of ICR/f rats during cataract development has not yet been established. In the present study, we determined changes in NO and LPO levels in lenses of ICR/f rats during cataract development. Opacification of ICR/f rat lenses started at 77 days of age, and the lenses of 91-day-old ICR/f rats were almost entirely opaque. The Ca(2+)-ATPase activity in the lenses of ICR/f rats decreased with increasing age, and an elevation in Ca(2+) content was observed in ICR/f rat lenses with the decrease in Ca(2+)-ATPase activity. NO levels in the lenses of ICR/f rats increased from 63 to 85 days of age, reaching a maximum at 77 days of age. In addition, LPO levels in the lenses of ICR/f rats also increased with increasing age. LPO levels in the lenses of 63- to 91-day-old ICR/f rats were found to be significantly higher compared with those in 22-day old ICR/f rats. These changes of Ca(2+), Ca(2+)-ATPase, NO and LPO were attenuated by instillation of DSF eye drops. These results suggest that excessive NO may cause enhanced lipid peroxidation resulting in the inhibition of Ca(2+) ATPase. The decrease in Ca(2+)-ATPase activity may cause the elevation in lens Ca(2+), leading to lens opacification in ICR/f rats. PMID- 18403085 TI - Co-oxidation-mediated xenobiotic activation and cytotoxicity by 12-lipoxygenase in intact platelets. AB - Peroxidase-mediated co-oxidation process has been suggested as a major alternative pathway for xenobiotic bioactivation other than hepatic cytochrome P450 monooxygenase system. Despite the wealth of reports on the possible involvement of co-oxidation in bioactivation of various compounds, clear manifestation of co-oxidation-mediated xenobiotic bioactivation in intact cell system without extra sources of peroxidase system has been difficult to demonstrate, mainly due to the natural scarcity of peroxidase activities in fresh intact cells. In the present study, arachidonic acid (AA) dependent bioactivation of alpha-naphthol, a representative phenolic compound, was demonstrated as shown by covalent binding increases in alpha-naphthol concentration dependent manner in freshly prepared intact platelets, where two AA dependent representative peroxidases, 12-lipoxygenase (12-LOX) and prostaglandin H synthase (PHS) are abundantly expressed. Inhibitors of 12-LOX attenuated the covalent binding of alpha-naphthol while inhibitors of PHS were not effective, indicating the predominant role of 12-LOX in AA-initiated co-oxidation in intact platelets. In addition, free radical scavengers and thiol donors prevented effectively the bioactivation of alpha-naphthol, suggesting the involvement of naphthoxy radical or naphthoxy-derived radical generation. Notably, the co-oxidation process resulted in enhanced cytotoxicities of alpha-naphthol against platelets indeed, as observed by cellular membrane disturbance and mitochondrial membrane potential decrease. With these results, we believe that an important in vitro evidence of peroxidase-mediated xenobiotic activation was provided for understanding the toxicological implication of peroxidase-mediated co-oxidation in the xenobiotic bioactivation. PMID- 18403086 TI - Adenoviral-mediated up-regulation of Otos, a novel specific cochlear gene, decreases cisplatin-induced apoptosis of cultured spiral ligament fibrocytes via MAPK/mitochondrial pathway. AB - Previous reports have implicated Otos, a novel specific gene expressed by spiral ligament fibrocytes (SLFs) with unclear functions, as a protective gene for cochlea. However, whether Otos gene could protect SLFs against cisplatin (DDP) induced apoptosis remains largely unknown. In the present study, we utilized Adenoviral-mediated gene transfection to up-regulate Otos expression in cultured SLFs and further assessed the cell viability and apoptosis as well as possible MAPK and mitochondrial pathways. As expected, the data showed that Otos up regulation significantly decreased apoptosis of SLFs induced by DDP possibly through activation of ERK and partial inhibition of JNK and mitochondrial pathway but not p-38 pathway, suggesting Otos as a potential protective gene for cochlea and raising the possibility of Otos up-regulation as a promising approach to DDP induced deafness therapy. PMID- 18403087 TI - Direction of signal recording affects waveform characteristics of ventricular fibrillation in humans undergoing defibrillation testing during ICD implantation. AB - INTRODUCTION: In cardiac arrest due to prolonged ventricular fibrillation (VF), defibrillation is more likely to result in a perfusing rhythm if chest compressions are performed first. Furthermore, the VF waveform can predict the shockability of VF and thus automated external defibrillators (AEDs) are being designed to analyze the VF waveform to direct therapies. However, it is unknown whether the VF waveform is dependent on recording direction, which could be altered by incorrect placement of AED patches. MATERIALS AND METHODS: VF was induced in 26 patients with ischemic cardiomyopathy and 19 patients with dilated cardiomyopathy and recorded in six limb leads. Frequency characteristics (mean, median, dominant frequency, and bandwidth) were computed as well as amplitude based measures: amplitude spectral area (AMSA), slope, signal amplitude, and slope divide by signal amplitude (slope-amp). RESULTS: Frequency characteristics were similar in all leads. However, AMSA, slope, and signal amplitude were significantly affected (P<0.001) by lead. In particular, for ischemic cardiomyopathy patients, between leads I and II, AMSA varied from 29.4+/-3.2 to 49.3+/-4.6 mV Hz (mean+/-SEM, P<0.001) and slope varied from 1.5+/-0.2 to 2.4+/ 0.3 mV/s (P<0.001). Slope-amp was similar in all leads. There were no significant differences between ischemic and dilated cardiomyopathy patients. CONCLUSIONS: Amplitude measures of VF are significantly affected by limb lead ECG recording direction. This work suggests that AED patches must be correctly and consistently placed if amplitude-based measures are used to decide whether to deliver a defibrillatory shock. PMID- 18403088 TI - Emergent diagnosis of acute coronary syndromes: today's challenges and tomorrow's possibilities. AB - Prompt diagnosis and effective early management of acute coronary syndromes within the Emergency Department are imperative. Arguably the most important step in the management of the acute coronary syndromes is identifying the problem in the first place. This narrative review explores the significant but under recognised limitations to current diagnostic strategies and addresses both contemporary and possible future solutions in a rapidly evolving field. PMID- 18403089 TI - Sexual behavior and findings on laparoscopy or laparotomy in women with severe chronic pelvic pain. AB - OBJECTIVE: To identify features of sexual behavior in women with and without chronic pelvic pain. STUDY DESIGN: A group of 41 women with chronic pelvic pain of more than 6 months' duration and scheduled to undergo laparoscopy or laparotomy were surveyed to obtain information on their sexual activity. The results were compared with a control group of 86 women of similar age and socioeconomic status who underwent planned tubal ligation for permanent contraception. RESULTS: Women with pelvic pain initiated sexual relations at a later age and had a higher rate of dyspareunia than women in the control group. There were no significant differences between groups in any other characteristics of sexual behavior. CONCLUSION: Dyspareunia, the only feature of sexual relations that differed significantly between women with and without chronic pelvic pain, should be accorded greater attention as a painful symptom and not be dismissed as simply a type of sexual dysfunction. PMID- 18403090 TI - Editorial comment on: laparoscopic and robotic assisted radical cystectomy for bladder cancer: a critical analysis. PMID- 18403091 TI - Editorial comment on: laparoscopic and robotic assisted radical cystectomy for bladder cancer: a critical analysis. PMID- 18403092 TI - Editorial comment on: endoluminal isoproterenol irrigation decreases renal pelvic pressure during flexible ureterorenoscopy: a clinical randomized, controlled study. PMID- 18403093 TI - Preliminary results of a novel method to estimate the probability of prostate cancer in men with elevated serum PSA values. PMID- 18403094 TI - Editorial comment on: treatment of bladder pain syndrome/interstitial cystitis 2008: can we make evidence-based decisions? PMID- 18403095 TI - Editorial comment on: treatment of bladder pain syndrome/interstitial cystitis 2008: can we make evidence-based decisions? PMID- 18403096 TI - Editorial comment on: endoluminal isoproterenol irrigation decreases renal pelvic pressure during flexible ureterorenoscopy: a clinical randomized, controlled study. PMID- 18403097 TI - Editorial comment on: preliminary results of a novel method to estimate the probability of prostate cancer in men with elevated serum PSA values. PMID- 18403098 TI - Editorial comment on: treatment of bladder pain syndrome/interstitial cystitis 2008: can we make evidence based decisions? PMID- 18403099 TI - Treatment of bladder pain syndrome/interstitial cystitis 2008: can we make evidence-based decisions? AB - CONTEXT: Opinions on how to best treat bladder pain/interstitial cystitis are ambiguous. OBJECTIVE: To review previous and recent literature on this subject to assess the current state of evidence. EVIDENCE ACQUISITION: With important previous papers reviewed for the 2003 European Association of Urology guidelines as background, the PubMed database was searched and articles published in 2003 2007 were reviewed and relevant ones were selected for detailed study. EVIDENCE SYNTHESIS: A large number of studies describing a variety of quite dissimilar therapeutic principles were retrieved. The various methods and level of evidence are summarised in tables. Only pentosan polysulfate sodium (oral and intravesical), amitriptyline, hydroxyzine, cyclosporin A, intravesical dimethyl sulfoxide, transurethral resection of visible Hunner lesions, and major reconstructive surgery reached a high degree of recommendation. However, a number of pitfalls hamper evaluation of the available information; a crucial one is that our understanding of basic mechanisms causing bladder pain is fragmentary. So far, we are faced with a large variety of hypotheses although it is difficult to identify the most relevant ones. In this respect, we are not much helped by the recent literature because many studies have poor descriptions of patients or are of a pilot character, with no follow-up by larger trials. Controlled studies are rather scarce. On the other hand, some good-quality studies following up positive pilot trials end up with negative results. CONCLUSION: Perhaps the most significant problem concerns inclusion and exclusion criteria in bladder pain syndrome/interstitial cystitis studies. At this stage, it is not too easy to communicate the wide available expert knowledge to the general audience. More sophisticated standards, capable of being generally used, have to come. PMID- 18403100 TI - Laparoscopic and robotic assisted radical cystectomy for bladder cancer: a critical analysis. AB - CONTEXT AND OBJECTIVES: Interest in laparoscopic assisted radical cystectomy (LRC) and robotic assisted radical cystectomy (RRC) is increasing at select centers worldwide. In this update we present the recent worldwide experience and critically evaluate the role of minimally invasive radical surgery for patients with bladder cancer. EVIDENCE ACQUISITION: English-language literature between 1992 and 2007 was reviewed using the National Library of Medicine database and the following key words: laparoscopic, laparoscopic-assisted, robotic, robotic assisted, and radical cystectomy. Over 102 papers were identified, 48 of which were selected for this review on the basis of their contribution to advancing the field with regard to three criteria: (1) evolution of concepts, (2) development and refinement of techniques, and (3) intermediate- and long-term clinical outcomes. These were evaluated with respect to current techniques and perioperative, functional, and oncological outcomes. Our initial experience is also reported. EVIDENCE SYNTHESIS: Minimally invasive techniques can adequately achieve the extirpative aspects of LRC and extended template lymphadenectomy. At most institutions the reconstructive urinary diversion is now typically being performed extracorporeally through a minilaparotomy. Perioperative data indicate that minimally invasive techniques are associated with reduced blood loss, slightly increased operating time, and shorter hospital stay without any significant difference in postoperative complications compared with open surgery. Intermediate-term oncological outcomes appear to be comparable with the open approach. Worldwide experience continues to increase; >700 surgeries have already been performed. CONCLUSION: LRC or RRC with extracorporeally constructed urinary diversion is a safe and effective operation for appropriate patients with bladder cancer. Perioperative and functional outcomes are comparable with open surgery. More focus on extended lymphadenectomy is necessary to routinely achieve higher node yields. Surrogate and intermediate oncological outcomes are encouraging, and long-term assessment is ongoing. PMID- 18403101 TI - Editorial comment on: endoluminal isoproterenol irrigation decreases renal pelvic pressure during flexible ureterorenoscopy: a clinical randomized, controlled study. PMID- 18403102 TI - Endoluminal isoproterenol irrigation decreases renal pelvic pressure during flexible ureterorenoscopy: a clinical randomized, controlled study. AB - OBJECTIVES: Irrigation during ureterorenoscopic procedures causes increased pelvic pressure (PP), which may lead to intrarenal backflow with potential harmful consequences. This study aims to investigate PP response to intraluminal administration of isoproterenol (beta-agonist; ISO) during flexible ureterorenoscopy. METHODS: Twelve patients admitted for retrograde intrarenal stone surgery (RIRS) were included. Patients were randomized to (1) irrigation with saline (n=6) or (2) irrigation with ISO 0.1 microg/mL (n=6). Irrigation rate was standardized to 8 mL/min. A ureteral catheter was retrogradely placed in the renal pelvis for PP measurements. PP, heart rate (HR), and mean arterial pressure (MAP) were also measured. RESULTS: Baseline PP was 12.1+/-4mm Hg in the saline group and 10.3+/-4mm Hg in the ISO group (p=0.44). In the saline group, PP increased to a mean 33+/-12 mm Hg during ureterorenoscopy. In the ISO group, PP was a mean 19+/-3mm Hg (p=0.029). During endoscopy, PP peaks as high as 328 mm Hg were noted during saline irrigation. The number of pressure peaks above 50mm Hg was minimized dramatically during ISO irrigation (p=0.035). No systemic side effects to ISO irrigation were observed. CONCLUSION: For the first time, a randomized, controlled human study demonstrates that pharmacologic modulation of the ureter is possible during upper urinary tract endoscopy. The ability to relax ureteral tone during endoscopy may have clinical advantages. PMID- 18403103 TI - Neuropeptide Y signalling on hippocampal stem cells in health and disease. AB - Neuropeptides are emerging as key components in the hippocampal neurogenic niche in health and disease, regulating many aspects of neurogenesis and the synaptic integration of newly generated neurons. This review focuses on the role of neuropeptide Y in the control of stem/precursor cells in the postnatal and adult hippocampus. It is likely that neuropeptide Y releasing interneurons are key sensors of neural activity, modulating neurogenesis appropriately. This is likely to be a fruitful area of research for extending our understanding of the control of stem cells in the normal and diseased brain. PMID- 18403104 TI - Hormone and growth factor signaling in endometrial renewal: role of stem/progenitor cells. AB - The human endometrium is a dynamic remodeling tissue undergoing more than 400 cycles of regeneration, differentiation and shedding during a woman's reproductive years. The co-ordinated and sequential actions of estrogen and progesterone direct these major remodeling events preparing a receptive endometrium for blastocyst implantation on a monthly basis. Adult stem/progenitor cells are likely responsible for endometrial regeneration. Functional approaches have been used to identify candidate endometrial stem/progenitor cells, as there are no specific stem cell markers. Rare populations of human endometrial epithelial and stromal colony-forming cells/units (CFU) and side population (SP) cells have been identified. Several growth factors are required for CFU activity: epidermal growth factor (EGF), transforming growth factor alpha (TGFalpha) and platelet-derived growth factor BB (PDGF-BB) for both epithelial and stromal CFU, and basic fibroblast growth factor (bFGF) for stromal, but not epithelial CFU. A sub-population of human endometrial stromal cells with mesenchymal stem cell properties of CFU activity and multilineage (fat, muscle, cartilage and bone) differentiation have been isolated by their co-expression of CD146 and PDGF receptor beta. Candidate epithelial and stromal stem/progenitor cells have been identified in mouse endometrium as rare label retaining cells (LRCs) in the luminal epithelium and as perivascular cells at the endometrial-myometrial junction, respectively. While epithelial and most stromal LRC do not express estrogen receptor alpha (Esr1), they rapidly proliferate on estrogen stimulation, most likely mediated by neighbouring Esr1-expressing niche cells. It is likely that these newly identified endometrial stem/progenitor cells may play key roles in the development of gynecological diseases associated with abnormal endometrial proliferation such as endometriosis and endometrial cancer. PMID- 18403105 TI - Androgen receptor signalling in prostate: effects of stromal factors on normal and cancer stem cells. AB - The prostate gland is the most common site for cancer in males within the developed world. Androgens play a vital role in prostate development, maintenance of tissue function and pathogenesis of prostate disease. The androgen receptor signalling pathway facilitates that role in both the epithelial compartment and in the underlying stroma. Stroma is a key mediator of androgenic effects upon the epithelium and can regulate both the fate of the epithelial stem cell and potentially the initiation and progression of prostate cancer. Different groups of growth factors are expressed by stroma, which control proliferation, and differentiation of prostate epithelium demonstrating a critical role for stroma in epithelial growth and homeostasis. Paracrine stromal proteins may offer the possibility to control tumour stem cell growth and could permit prostate specific targeting of both therapies and of androgen responsive proteins. The effect of 5alpha-dihydrotestosterone, the more potent metabolite of testosterone, on expression of androgen-regulated genes in stroma from benign prostatic hyperplasia is a key mediator of epithelial cell fate. Global gene expression arrays have recently identified new candidate genes in androgen responsive stroma, some of which have androgen receptor binding sites in their promoter regions. Some of these genes have direct androgen receptor binding ability. PMID- 18403106 TI - New insights into type II germ cell tumor pathogenesis based on studies of patients with various forms of disorders of sex development (DSD). AB - Disorders of sex development (DSD), previously known as intersex, refer to congenital conditions in which development of chromosomal, gonadal, or anatomical sex is atypical. Patients with specific variants of this disorder have an elevated risk for the development of so-called type II germ cell cancers, i.e., the seminomatous and nonseminatous tumors, referred to as germ cell tumors (GCTs). Specifically DSD patients with gonadal dysgenesis or hypovirilization are at risk. A prerequisite for type II GCT formation is the presence of a specific part of the Y chromosome (referred to as the GBY region), with the TSPY gene being the most likely candidate. Also the octamer binding transcription factor OCT3/4 is consistently expressed in all type II GCTs with pluripotent potential, as well as in the precursor lesions carcinoma in situ (CIS) in case of a testis and gonadoblastoma (GB) in the DSD gonad. The actual risk for malignant transformation in individual DSD patients is hard to predict, because of confusing terminology referring to the different forms of DSD, and unclear criteria for identification of the presence of malignant germ cells, especially in young patients. This is specifically due to the phenomenon of delay of germ cell maturation, which might result in over diagnosis. This review will give novel insight into the pathogenesis of the type II GCTs through the study of patients with various forms of DSD for which the underlying molecular defect is known. To allow optimal understanding of the pathogenesis of this type of cancers, first normal gonadal development, especially regarding the germ cell lineage, will be discussed, after which type II GCTs will be introduced. Subsequently, the relationship between type II GCTs and DSD will be described, resulting in a number of new insights into the development of the precursor lesions of these tumors. PMID- 18403107 TI - Life in the crypt: a role for glucagon-like peptide-2? AB - The epithelial layer of the intestinal tract serves as a model to study the mechanisms regulating tissue renewal. Central to this process is the intestinal stem cell and, thus, both the intrinsic and extrinsic factors that modulate the function of these cells must be understood. Amongst the intrinsic regulators, both the canonical wnt and bone morphogenic protein (bmp) signaling pathways have been shown to be essential determinants of stem cell dynamics and intestinal homeostasis. The intestinotrophic hormone, glucagon-like peptide-2 (GLP-2), has also recently been demonstrated to exert a variety of effects on the intestinal crypt cells, including enhancement of the putative stem cell marker, musashi-1, as well as stimulating intestinal proliferation. As the GLP-2 receptor is not expressed by the crypt cells, these actions have been hypothesized to be mediated indirectly, through other gut peptides and/or growth factors. Of these, recent studies have demonstrated a requirement for insulin-like growth factor-1 in the proliferative effects of GLP-2, through a pathway that involves activation of the canonical wnt signaling pathway. This extrinsic pathway represents a novel mechanism by which intestinal stem cell dynamics may be regulated. PMID- 18403108 TI - Viscosity of high-alcohol content fuel blends with water: subsurface contaminant transport implications. AB - In the United States, a gasoline fuel blend with alcohol volume fractions of 85% or more is considered (among other fuels) an alternative fuel. As the popularity and usage of high-alcohol content gasoline fuel blends increase, subsurface contamination from these fuels will be of great environmental concern. An important parameter governing the movement of these contaminants in unsaturated porous media is the liquid viscosity. In this study, five sets of experiments are conducted to determine viscosity variations of (a) blends of 15% gasoline with various alcohol mixtures, and (b) mixtures of high-alcohol content gasoline fuel blends with various volume fractions of water. The three alcohols considered in this study are ethanol, methanol, and isopropanol. The viscosity of each liquid mixture is observed using a modified falling-ball viscometer. PMID- 18403109 TI - Kinetic studies for the biosorption of lead and copper ions by Penicillium simplicissimum immobilized within loofa sponge. AB - Biosorption potential of Penicillium simplicissimum (Penicillium sp.) immobilized within loofa sponge (PSILS) for lead and copper from aqueous media was explored. The effects of pH, contact time and initial concentration were studied in batch experiments. The maximum uptake of metal ions was obtained at pH 5.0. Biosorption equilibrium was established by 60 min. The pseudo-first-order, pseudo-second order and intraparticle diffusion kinetic models were applied to study the kinetics of the biosorption processes. The pseudo-second-order kinetic model provided the best correlation (R(2)>0.999) of the experimental data compared to the pseudo-first-order and intraparticle diffusion kinetic models. The maximum heavy metal ions adsorbed was found to be 152.6 and 112.3mg/g for Pb(II) and Cu(II), respectively. It was found that the biosorption of both Pb(II) and Cu(II) on PSILS was correlated well (R(Pb)(2)=0.999 and R(Cu)(2)=0.9978) with the Langmuir equation as compared to Freundlich isotherm equation under the concentration range studied. PSILS was regenerated by washing with a 100mM solution of HCl. The desorption efficiency was as high as 98%. The PSILS was reused in five adsorption-desorption cycles with negligible decrease in biosorption capacity. The present work showed that PSILS was an efficient biosorbent for removal of heavy metal ions from aqueous solution. PMID- 18403110 TI - Effect of surface acidic oxides of activated carbon on adsorption of ammonia. AB - The influence of surface acidity of activated carbon (AC) was experimentally studied on adsorption of ammonia (NH(3)). Coconut shell-based AC was modified by various acids at different concentrations. There were five different acids employed to modified AC, which included nitric acid, sulfuric acid, hydrochloric acid, phosphoric acid, and acetic acid. Acidic functional groups on the surface of ACs were determined by a Fourier transform infrared spectrograph (FTIR) and by the Boehm titration method. Specific surface area and pore volume of the ACs were measured by a nitrogen adsorption apparatus. Adsorption amounts of NH(3) onto the ACs were measured by a dynamic adsorption system at room temperature according to the principle of the ASTM standard test method. The concentration of NH(3) in the effluent stream was monitored by a gas-detecting tube technique. Experimental results showed that adsorption amounts of NH(3) on the modified ACs were all enhanced. The ammonia adsorption amounts on various activated carbons modified by different acids are in the following order: nitric acid>sulfuric acid>acetic acid approximately phosphoric acid>hydrochloric acid. It is worth to note that the breakthrough capacity of NH(3) is linearly proportional to the amount of acidic functional groups of the ACs. PMID- 18403111 TI - Isolation and characterization of a nitrobenzene degrading yeast strain from activated sludge. AB - Strain Z1 was isolated from nitrobenzene-contaminated sludge. Strain Z1 was able to utilize nitrobenzene as a sole source of carbon, nitrogen, and energy under aerobic condition. Based on the morphology, physiological biochemical characteristics, and 26S rDNA D1/D2 domain sequence, strain Z1 was identified as Rhodotorula mucilaginosa. Strain Z1 mineralized up to 450mg L(-1) nitrobenzene. Kinetics of nitrobenzene degradation was described using the Andrews equation. The kinetic parameters were as follows: q(max)=1.50h(-1), K(s)=31.31mg L(-1), and K(i)=101.34mg L(-1). Strain Z1 had a high-salinity tolerance. It degraded nitrobenzene effectively in 5% NaCl (quality concentration). Even in the presence of aniline or phenol, strain Z1 degraded nitrobenzene efficiently. Strain Z1 therefore could be an excellent candidate for the bio-treatment of nitrobenzene industrial wastewaters. PMID- 18403112 TI - Inhibition of biogas production and biodegradability by substituted phenolic compounds in anaerobic sludge. AB - Phenolic compounds are abundant in nature and organic wastes. This biomass may be utilised in biogas generation. Phenolics can inhibit the degradation of readily biodegradable organic fractions and their own biodegradation. In this work, assays were carried out under anaerobic conditions to study the inhibition of both gas production and biodegradability due to seven phenolic compounds and to study their adsorption onto sludge and autoxidation in the aqueous medium. Fifty percent inhibition was in the range of 120 to 594 mg of compound/g VSS. An initial enhancement followed by an inhibition of biogas formation was found. The inhibition by the phenolic compounds was found to be influenced by autoxidation, apolarity, type, size and number of substitutions. Biogas production is influenced by concentration rather than any pH change. The concentration of the phenolic compound was partially biomethanized and the degradation of gallic and caffeic acids by this process is reported here for the first time. The maximum total biodegradation of any phenolic compound was 63.85+/-2.73%, and remaining non-biodegradable fraction was autoxidized and adsorbed onto the sludge matrix. Inhibition of methanization and partial inhibition of background gas was found at concentrations between 800 and 1600 mg/L organic carbon. PMID- 18403113 TI - Involvement of group I mGluRs in LTP induced by strong high frequency stimulation in the dentate gyrus in vitro. AB - The involvement of group I metabotropic glutamate receptors (mGluRs) and ryanodine receptors was investigated in the induction of LTP induced either by application of one standard high frequency stimulation (HFS) or by strong multiple HFS in the medial perforant path to granule cell synapse of the rat dentate gyrus. Whilst a standard brief HFS induced LTP close to 50%, strong stimulation consisting of multiple HFS induced a much larger LTP. mGluR5 was found to be partially involved in the induction of the enhanced LTP induced by the strong HFS but not in the standard LTP induced by the brief HFS. Thus the mGluR5 antagonists LY341495 and MPEP partially inhibited the induction of LTP induced by strong HFS but did not inhibit LTP induced by a standard HFS. Ryanodine was found to partially inhibit LTP induced by the strong HFS but not to inhibit the standard LTP induced by the brief HFS, demonstrating the involvement of Ca-induced Ca release from ryanodine-sensitive Ca stores in the former. These studies demonstrate that the large amplitude LTP induced by strong stimulation involves additional mechanisms to the LTP induced by brief HFS, in particular involving activation of mGluR5 and RyR-sensitive Ca stores. PMID- 18403114 TI - APPswe mutation increases the frequency of spontaneous Ca2+-oscillations in rat hippocampal neurons. AB - Altered calcium homeostasis is implicated in the pathogenesis of Alzheimer's disease (AD). Much effort has been put into understanding the association between protein mutations causative of this devastating neurodegenerative disease and perturbed calcium signaling. Whereas the presenilin mutations have received most attention in the context of neuronal calcium signaling, we focused on the effects of APP with the so-called Swedish mutation (APPswe) on spontaneous neuronal activity. We observed that primary hippocampal neurons from an APPswe transgenic rat showed increased frequency and unaltered amplitude of spontaneous calcium oscillations as compared to wild-type neurons. We found that the altered calcium signaling of APPswe transgenic neurons was unlikely to be due to modulation of the NMDA or nicotinic neurotransmitter systems, and did not depend on secreted APP derivates. The implications of this effect of APP are discussed. PMID- 18403115 TI - Cytokine regulation in animal models of neuropathic pain and in human diseases. AB - Cytokines are soluble proteins secreted mainly by immune cells and are key players in the induction and maintenance of pain. Pro-inflammatory cytokines are mostly algesic, while anti-inflammatory cytokines have analgesic properties. After the role of cytokines was shown in diverse animal models of pain, interest arose in the systemic and local regulation of cytokines in human pain states. Most clinical studies give evidence for an imbalance between pro- and anti inflammatory cytokines in neuropathic and other pain states with pronounced pro inflammatory cytokine profiles. Anti-cytokine treatment gives encouraging preliminary results and supports the notion of a crucial role of cytokines also in human pain states. Further research is needed for a better understanding of the mechanisms linking altered cytokine profiles to the sensation of pain. PMID- 18403116 TI - Gene therapy directed at the neuroimmune component of chronic pain with particular attention to the role of TNF alpha. AB - Identification that neuroimmune activation in the spinal cord is an important factor in the development of chronic pain has opened the possibility that gene transfer of anti-inflammatory peptides may be used to reduce pain neurotransmission. We review the published evidence regarding gene transfer to meninges to express the anti-inflammatory peptide interleukin 10, and gene transfer to dorsal root ganglia using replication incompetent HSV vectors to express interleukin 4, interleukin 10, or the soluble (p55) tumor necrosis factor receptor (sTNFR). The results of these experiments suggest a novel role for "reverse signaling" through the full-length membrane form of TNFalpha in spinal glia in the modulation of chronic pain. PMID- 18403117 TI - Acute stress increases neuropsin mRNA expression in the mouse hippocampus through the glucocorticoid pathway. AB - Stress affects synaptic plasticity and may alter various types of behaviour, including anxiety or memory formation. In the present study, we examined the effects of acute stress (1 h restraint with or without tail-shock) on mRNA levels of a plasticity-related serine protease neuropsin (NP) in the hippocampus using semiquantitative RT-PCR and in situ hybridization. We found that NP mRNA expression was dramatically increased shortly after exposure to the acute restraint tail-shock stress and remained at high level for at least 24 h. The level of NP mRNA would be correlated to the elevated plasma concentration of the glucocorticoid corticosterone (CORT) and to the stress intensity. Application of CORT either onto primary cultured hippocampal neurons (5 nM) or in vivo to adrenalectomized (ADX) mice (10 mg/kg B.W., s.c.) mimicked the effect of stress and significantly elevated NP mRNA. These results suggest that the upregulation of NP mRNA after stress is CORT-dependent and point to a role for neuropsin in stress-induced neuronal plasticity. PMID- 18403118 TI - Cyclooxygenase-2 is involved in oxidative damage and alpha-synuclein accumulation in dopaminergic cells. AB - Cyclooxygenase (COX) is the rate-limiting enzyme that catalyzes the formation of prostaglandins from arachidonic acid. The inducible isoform COX-2 is upregulated in the dopaminergic neurons of the substantia nigra of postmortem Parkinson's disease (PD) patients and in neurotoxin-induced Parkinsonism models. COX-2 has attracted significant attention as an important source of oxidative stress in dopaminergic neurons due to its potential to oxidize catechols including dopamine. However, the role of COX-2 in the pathogenesis of PD has not been fully evaluated. Here, we show that COX-2 induces dopamine oxidation, as evidenced by the findings that COX-2 can facilitate dopamine oxidation in a cell-free system and in COX-2-overexpressing SH-SY5Y cells, and that this can be completely abolished by the selective COX-2 inhibitor meloxicam. Increased COX-2 expression causes oxidative protein modification and alpha-synuclein accumulation in dopaminergic cells. These data suggest that an abnormal increase in COX-2 expression causes dopamine oxidation and contributes to the preferential vulnerability of dopaminergic cells as in PD. PMID- 18403119 TI - Promoting of wakefulness by administrations of modafinil into anterior hypothalamus and into the pedunculopontine tegmental nucleus in rats. AB - We investigated whether administration of MOD in rats during the lights-on period into wake-promoting areas, such as anterior hypothalamus (AH) or into the pedunculopontine tegmental nucleus (PPTg) would enhance waking. Results showed that microinjections of 1 microL of MOD (10, 20, or 30 microg) into both brain areas increased the total time of alertness and decreased sleep. Additionally, MOD-treated rats showed an enhancement in alpha power spectra but delta power spectra was diminished. Finally, c-Fos expression was found increased into either AH or the PPTg. Collectively, these results suggest that MOD induces waking via the activity of two wake-related brain areas such as AH and the PPTg. PMID- 18403120 TI - Effects of chewing in working memory processing. AB - It has been generally suggested that chewing produces an enhancing effect on cognitive performance-related aspects of memory by the test battery. Furthermore, recent studies have shown that chewing is associated with activation of various brain regions, including the prefrontal cortex. However, little is known about the relation between cognitive performances affected by chewing and the neuronal activity in specified regions in the brain. We therefore examined the effects of chewing on neuronal activities in the brain during a working memory task using fMRI. The subjects chewed gum, without odor and taste components, between continuously performed two- or three-back (n-back) working memory tasks. Chewing increased the BOLD signals in the middle frontal gyrus (Brodmann's areas 9 and 46) in the dorsolateral prefrontal cortex during the n-back tasks. Furthermore, there were more prominent activations in the right premotor cortex, precuneus, thalamus, hippocampus and inferior parietal lobe during the n-back tasks after the chewing trial. These results suggest that chewing may accelerate or recover the process of working memory besides inducing improvement in the arousal level by the chewing motion. PMID- 18403121 TI - Effects of MK-886, a 5-lipoxygenase activating protein (FLAP) inhibitor, and 5 lipoxygenase deficiency on the forced swimming behavior of mice. AB - A common biological pathway may contribute to the comorbidity of atherosclerosis and depression. Increased activity of the enzymatic 5-lipoxygenase (5-LOX, 5LO) pathway is a contributing factor in atherosclerosis and a 5-LOX inhibitor, MK 886, is beneficial in animal models of atherosclerosis. In the brain, MK-886 increases phosphorylation of the glutamate receptor subunit GluR1, and the increased phosphorylation of this receptor has been associated with antidepressant treatment. In this work, we evaluated the behavioral effects of MK 886 in an automated assay of mouse forced swimming, which identifies antidepressant activity as increased climbing behavior and/or decreased rest time. Whereas a single injection of MK-886 (3 and 10 mg/kg) did not affect forced swimming behaviors assayed 30 min later, six daily injections of 3 mg/kg MK-886 slightly increased climbing and significantly reduced rest time in wild-type mice but not in 5-LOX-deficient mice. A diet delivery of MK-886, 4 micro/(100 mg(body weight)day), required 3 weeks to affect forced swimming; it increased climbing behavior. Climbing behavior was also increased in naive 5-LOX-deficient mice compared to naive wild-type controls. These results suggest that 5-LOX inhibition and deficiency may be associated with antidepressant activity. Increased climbing in a forced swimming assay is a typical outcome of antidepressants that increase noradrenergic and dopaminergic activity. Interestingly, 5-LOX deficiency and MK 886 treatment have been shown to be capable of increasing the behavioral effects of a noradrenaline/dopamine-potentiating drug, cocaine. Future research is needed to evaluate the clinical relevance of our findings. PMID- 18403123 TI - Bloch wave analysis of the Eshelby twist contrast around end-on screw dislocations in bcc Mo. AB - In high-resolution electron microscopy (HREM), dislocation core structures are examined by tilting the dislocation end-on along the appropriate zone axis. For end-on screw dislocations diffraction contrast is largely due to surface relaxation in the form of the Eshelby twist. In this paper, simulated, many-beam images of end-on, 1/2<111> Mo screw dislocations in a thin TEM foil are presented. The diffraction contrast is found to be rotationally symmetric and of very low value. Bloch waves are used to explain the physical origin of the contrast. Diffraction contrast is, however, significantly enhanced by using an annular aperture which includes only the diffracted beams. Annular aperture contrast is largest for foil thicknesses corresponding to minima in the dark field pendellosung. PMID- 18403124 TI - Galactorrhea and amenorrhea in burn patients. AB - INTRODUCTION: Galactorrhea and/or amenorrhea, although uncommonly reported in post-burn patients, is a complex problem to treat. Patient is reluctant to volunteer history of these symptoms, unless asked specifically. AIM: To study profile of adult female patients with galactorrhea and/or amenorrhea in post burn period. MATERIALS AND METHOD: A prospective study of all adult female patients presenting with or detected to have galactorrhea and/or amenorrhea in post burn period was conducted over 6 month's period. Detailed clinical examination, estimation of LH, FSH, Prolactin levels and X-ray of skull was done in all patients. The data collected was analyzed. Patients with hyperprolactinemia and galactorrhea were treated with Bromocriptine for 3 weeks to 3 months. In all patients with amenorrhea, pregnancy was ruled out by gynecological examination and urine pregnancy test. RESULTS: During this period, 30 patients (15.15%) were detected to have galactorrhea and/or amenorrhoea. The extent of burn in these patients was 20-65%of body surface area. Out of 30 patients, 5 had galactorrhea and amenorrhea, 1 galactorrhea alone and 24 had amenorrhea alone. Analysis of voluntary disclosures and detection on interrogation was done. Till the end of study, 4 patients with galactorrhea had complete relief, 2 patients reported reduction in discharge. DISCUSSION: Galactorrhea was distressing for all and was always associated with high prolactine levels .The reverse was not true. All the patients had chest burns besides other body areas. Association was noted between menstrual aberration and ovulatory phase at the time of burn. CONCLUSION: Galactorrhea and menstrual disturbances do exist in female patients in reproductive age group in post burn period and patients should be especially interrogated for these symptoms by the burn care providers. PMID- 18403122 TI - Genetic variability of the mu-opioid receptor influences intrathecal fentanyl analgesia requirements in laboring women. AB - Labor initiates one of the most intensely painful episodes in a woman's life. Opioids are used to provide analgesia with substantial interindividual variability in efficacy. mu-Opioid receptor (muOR, OPRM1) genetic variants may explain differences in response to opioid analgesia. We hypothesized that OPRM1 304A/G polymorphism influences the median effective dose (ED(50)) of intrathecal fentanyl via combined spinal-epidural for labor analgesia. Nulliparous women were prospectively recruited around 35 weeks gestation (n=224), and genotyped for 304A/G polymorphism. Those requesting neuraxial labor analgesia were enrolled in one of the two double-blinded trials: up-down sequential allocation (SA, n=50) and a separate confirmatory random-dose allocation trial (RA, n=97). Effective analgesia from intrathecal fentanyl was defined by >or=60 min analgesia with verbal rating score 2nmol/l) were significantly higher after the SECPT. Social evaluation during hand immersion in warm water did not affect saliva cortisol levels suggesting that both social evaluation and a challenge are required for HPA axis activation. These findings indicate that the incorporation of social-evaluative elements increases HPA axis responses to the CPT. The SECPT can serve as a tool for future stress research. PMID- 18403131 TI - Pill-poppers and dopers: a comparison of non-medical prescription drug use and illicit/street drug use among college students. AB - Data from the 2001 College Alcohol Study, a national sample of U.S. college students, were used to conduct multinomial logistic regression analysis examining correlates of substance use. Students were divided into three groups based on their lifetime substance use: non-users, non-medical prescription drug use only, and illicit/street drug use only. The purpose of this analytic strategy was to examine the similarities/differences in the correlates of non-medical prescription drug use and illicit/street drug use. Findings indicate that race, age, G.P.A., sexual activity, health, binge drinking, marijuana use, social bonding and social learning measures are correlates of non-medical prescription drug use. Correlates of illicit/street drug use include gender, Hispanic ethnicity, sexual activity, binge drinking, marijuana use, social bonding and social learning measures. Finally, the focus of the paper is a comparison of students who report only non-medical prescription drug use to students who report only illicit/street drug use. Findings indicate that gender, race, marital status, sexual activity, marijuana use, and social bonding measures significantly distinguish illicit/street drug use from non-medical prescription drug use. Important implications, limitations, and future research needs were discussed. PMID- 18403132 TI - Substance use disorders among inhalant users: results from the National Epidemiologic Survey on alcohol and related conditions. AB - OBJECTIVE: To assess the prevalence, correlates, and age of onset of DSM-IV substance use disorders (SUDs) among adult inhalant users. METHODS: Analyses were based on structured psychiatric interviews of a nationally representative sample of 43,093 US adults. RESULTS: The lifetime prevalence of SUDs among adult inhalant users was 96%. Alcohol (87%), marijuana (68%), nicotine (58%), cocaine (35%), hallucinogen (31%), and stimulant (28%) use disorders were more prevalent than inhalant use disorders (19%). An estimated 62% of inhalant users met criteria for a past-year SUD. Less education, residence in non-metropolitan areas, early onset of inhalant use, and a history of substance abuse treatment were associated with increased odds of having an inhalant use disorder. Inhalant users who were under age 30 or who were members of families with low incomes had increased odds of having nicotine dependence and an alcohol or drug use disorder in the past year. Compared with substance users without a history of inhalant use, inhalant users, on average, initiated use of cigarettes, alcohol, and almost all other drugs at younger ages, and had a higher lifetime prevalence of nicotine, alcohol, and any drug use disorder. CONCLUSIONS: Lifetime and past-year SUDs are prevalent among adults with a history of inhalant use. PMID- 18403133 TI - Long-term outcomes of radiotherapy for pituitary adenomas. AB - PURPOSE: To evaluate long-term local control and toxicity for pituitary adenomas treated with fractionated radiotherapy (RT). METHODS AND MATERIALS: The records of 100 patients with pituitary adenomas treated between 1983 and 2003 were retrospectively reviewed. Thirty-one patients had hormone-secreting tumors; 69 patients were treated with surgery and postoperative RT. Median follow-up was 6.7 years (range, 0.6-20.2 years) for all patients and 6.2 years (range, 2-20.2 years) for living patients. The mean dose delivered was 45 Gy (range, 43-50.4 Gy). RESULTS: The 10-year actuarial local control rates for nonsecreting and secreting adenomas were 98% and 73%, respectively (p = 0.0015). Actuarial 10-year cause-specific survival (CSS) rates were 95% and 88%, and overall survival rates were 66% and 79% for nonsecreting and secreting adenomas, respectively. Involvement of the sphenoid sinus was found to be significantly associated with decreased 10-year CSS (p = 0.0453). When compared with the two- or three-field techniques, stereotactic RT was associated with improved CSS (p = 0.0775). CSS was not significantly associated with hormone excretion, extent of surgery, or whether RT was administrated postoperatively or for salvage after a postsurgical recurrence. New cases of hypopituitarism occurred in 35 patients. One patient experienced vision loss, and one patient developed a post-treatment glioma. CONCLUSIONS: This is one of the most mature series in the literature that documents excellent results with fractionated RT for pituitary adenoma. We recommend 45 Gy at 1.8 Gy per fraction using stereotactic noncoplanar fields. PMID- 18403134 TI - Atomoxetine treatment of adults with ADHD and comorbid alcohol use disorders. AB - OBJECTIVE: Adults with attention-deficit/hyperactivity disorder (ADHD) have higher rates of alcohol and drug use disorders than adults without ADHD. The study aim was to determine if atomoxetine was superior to placebo in improving ADHD and alcohol use in recently abstinent adults with ADHD and comorbid alcohol use disorder. METHODS: Adults with DSM-IV diagnoses of ADHD and alcohol abuse and/or dependence were abstinent from alcohol at least 4 days (maximum 30 days) before study randomization. Participants received atomoxetine (25-100mg daily) or placebo for 12 weeks. ADHD symptoms were assessed using ADHD Investigator Symptom Rating Scale (AISRS) total score. Time-to-relapse to heavy alcohol use was analyzed using a 2-sided log-rank test based on Kaplan-Meier estimates and cumulative heavy drinking events over time were evaluated post hoc with recurrent event analysis. RESULTS: Subjects received atomoxetine (n=72) or placebo (n=75) and 80 subjects completed the 12-week double-blind period (n=32 and 48, respectively). ADHD symptoms were significantly improved in the atomoxetine cohort compared to placebo (AISRS total score mean [S.D.], atomoxetine: -13.63 [11.35], P<.001; placebo: -8.31 [11.44], P<.001, difference: P=.007; effect size=0.48). No significant differences between treatment groups occurred in time to-relapse of heavy drinking (P=.93). However, cumulative heavy drinking days were reduced 26% in atomoxetine-treated subjects versus placebo (event ratio=0.74, P=.023). There were no serious adverse events or specific drug-drug reactions related to current alcohol use. CONCLUSIONS: This 3-month, double blind, placebo-controlled study of atomoxetine in adults with ADHD and comorbid alcohol use disorder demonstrates clinically significant ADHD improvement, and inconsistent effects on drinking behavior. PMID- 18403135 TI - Identification and characterization of a novel folliculin-interacting protein FNIP2. AB - Birt-Hogg-Dube' syndrome characterized by increased risk for renal neoplasia is caused by germline mutations in the BHD/FLCN gene encoding a novel tumor suppressor protein, folliculin(FLCN), which interacts with FNIP1 and 5'-AMP activated protein kinase(AMPK). Here we report the identification and characterization of a novel FNIP1 homolog FNIP2 that also interacts with FLCN and AMPK. C-terminally-deleted FLCN mutants, similar to those produced by naturally occurring germline mutations in BHD patients, were unable to bind FNIP2. These data taken together with our previous results that demonstrated FNIP1 binding to the C-terminus of FLCN suggest that FLCN tumor suppressor function may be facilitated by interactions with both FNIP1 and FNIP2 through its C-terminus. Furthermore, we demonstrate that FNIP1 and FNIP2 are able to form homo- or heteromeric multimers suggesting that they may function independently or cooperatively with FLCN. Differential expression of FNIP1 and FNIP2 transcripts in some normal tissues may indicate tissue specificity for these homologs. Interestingly FNIP1 and FNIP2 were oppositely expressed in human clear cell renal cell carcinoma (RCC), and coordinately expressed in chromophobe RCC and oncocytoma, suggesting their differential function in different histologic variants of RCC. PMID- 18403136 TI - High genetic diversity in RdRp gene of Brazilian porcine sapovirus strains. AB - Sapovirus is one genus within Caliciviridae family that causes diarrhea in humans and animals. Sapovirus (SaV) has been classified into seven genogroups (GI to GVII). The GIII, GVI, and GVII, which prototype is Cowden, JJ681, and K7/JP strains, respectively, infect pigs. The objective of this study was to characterize wild-type Brazilian SaV strains from piglet stool samples and determine SaV infection frequency, age distribution and association with diarrheic disease. Stool samples from 113 piglets up to 28-days-old were collected from 34 pig farms located in the States of Minas Gerais (MG), Mato Grosso do Sul (MS), Parana (PR), Santa Catarina (SC), and Rio Grande do Sul (RS), during 2004 and 2005. The specimens were evaluated for enteric calicivirus by RT PCR assay with primers p289/290, designed to detect the polymerase gene of SaV and norovirus. Thirty four (30.1%) samples were positive for SaV and five amplicons were sequenced. Phylogenetic analyses placed BRA29-MS/04 and BRA52 PR/05 sequences into the GIII of SaV genus. BRA04-SC/04, BRA21-RS/04, and BRA37 MG/05 demonstrated low identity with the Cowden strain but were closely related (up to 86.3%) to the Japanese and Dutch SaV strains, grouping together in a new cluster (GVIII?) in the phylogenetic tree. SaV infection was detected more frequently (p=0.0001) in animals between 22 and 28 days of age, in equal frequencies in piglets with and without diarrhea (p=0.59), and in the five Brazilian States. In this study, such as other unclassified worldwide SaVs, the Brazilian strains showed high genetic variability. Furthermore, the distribution and frequency of SaV infection provides evidence that the virus is circulating in Brazilian pig herds. PMID- 18403137 TI - Isolation and genetic characterization of avian origin H9N2 influenza viruses from pigs in China. AB - As pigs are susceptible to infection with both avian and human influenza A viruses, they have been proposed to be an intermediate host for the adaptation of avian influenza viruses to humans. In April 2006, a disease caused by highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) occurred in several pig farms and subsequently overwhelmed almost half of China with more than 2,000,000 cases of pig infection. Here we report a case in which four swine H9N2 influenza viruses were isolated from pigs infected by highly pathogenic PRRSVs in Guangxi province in China. All the eight gene segments of the four swine H9N2 viruses are highly homologous to A/Pigeon/Nanchang/2-0461/00 (H9N2) or A/Wild Duck/Nanchang/2-0480/00 (H9N2). Phylogenetic analyses of eight genes show that the swine H9N2 influenza viruses are of avian origin and may be the descendants of A/Duck/Hong Kong/Y280/97-like viruses. Molecular analysis of the HA gene indicates that our H9N2 isolates might have high-affinity binding to the alpha2,6-NeuAcGal receptor found in human cells. In conclusion, our finding provides further evidence about the interspecies transmission of avian influenza viruses to pigs and emphasizes the importance of reinforcing swine influenza virus (SIV) surveillance, especially after the emergence of highly pathogenic PRRSVs in pigs in China. PMID- 18403138 TI - Cloning, expression and characterization of a cell wall surface protein, 6 phosphogluconate-dehydrogenase, of Streptococcus suis serotype 2. AB - Streptococcus suis type 2 is a pathogen responsible for diverse diseases in both pigs and humans. In order to understand the pathogenesis of the S. suis type 2 infection, the gene encoding a cell surface protein, 6-phosphogluconate dehydrogenase (6PGD) of S. suis type 2 was cloned and sequenced, and recombinant 6PGD protein (r6PGD) was produced in a prokaryotic expression system. Sequence analysis of the cloned 6 pdg gene showed 82% similarity with Streptococcus pneumoniae 6 pdg at the nucleic acid level. Western blotting using r6PGD-specific antiserum confirmed the cell surface location of the 6PGD protein of S. suis type 2. The role of 6PGD in S. suis type 2 pathogenesis as an adhesin and its immunogenicity in mice was further investigated. The results showed that the recombinant protein interfered with the adhesion of S. suis type 2 to Hep2 and HeLa cells by 72% and 66%, respectively. Immunization of CD-1 mice with r6PGD increased the protective efficacy by 80% following intraperitoneal administration of a lethal dose of S. suis type 2. Immunization of CD-1 mice with r6PGD elicited a significant protective immune response, which demonstrated the importance of 6PGD to bacterial pathogenesis. Identification and characterization of the role of S. suis type 2 6PGD in adhesion and immunogenicity will allow us to use this protein to develop new antimicrobial therapies and/or vaccines. PMID- 18403139 TI - How is maternal recollection of the birth experience related to the behavioral and emotional outcome of preterm infants? AB - OBJECTIVES: To investigate how mother's recollections of birth experiences and first contact with the newborn relate to the child's behavioral and emotional problems at five to six years of age. METHODS: The study included 28 mothers of preterm (birth weight < or =2500 g) and 39 mothers of full-term children, born in Tampere University Hospital in 1998. When the children were five to six years old, maternal recollections of the birth experiences were assessed using the Clinical Interview for Parents of High-Risk Infants (CLIP) and children's behavioral and emotional problems were assessed using the Child Behavior Checklist (CBCL). RESULTS: Mothers of the preterm children still had more negative recollections of the labor (p < 0.001) and first contact with the newborn (p < 0.001) than mothers of the full-term children. These recollections related to the child's behavioral and emotional symptoms when the child was five to six years old in the preterm group but not in the full-term group. CONCLUSION: The impact of mother's birth experience seems to have long-lasting effects on the preterm child. This finding emphasizes the importance of early physical mother infant contact and supporting the mothers of preterm infants, especially if they articulate negative or traumatic experiences related to the birth of their child. PMID- 18403140 TI - Non-functionalized multi-walled carbon nanotubes alter the paracellular permeability of human airway epithelial cells. AB - Little information is available upon the effects of carbon nanotubes (CNT) on the airway barrier. Here we study the barrier function of Calu-3 human airway epithelial cells, grown on permeable filters, after the exposure to commercial single-walled or multi-walled CNT, produced through chemical vapour deposition. To assess changes in the paracellular permeability of CNT-treated Calu-3 monolayers, we have measured the trans-epithelial electrical resistance (TEER) and the permeability to mannitol. Multi-walled CNT caused a large decrease in TEER and an increase in mannitol permeability but no substantial alteration in monolayer viability. Single-walled CNT produced much smaller changes of TEER while CNT, synthesized through the arc discharge method, and Carbon Black nanoparticles had no effect. If commercial multi-walled CNT were added during the formation of the tight monolayer, no further increase in trans-epithelial resistance was observed. Moreover, the same nanomaterials, but neither single walled counterparts nor Carbon Black, prevented the TEER recovery observed after the discontinuation of interleukin-4, a Th2 cytokine that causes a reversible barrier dysfunction in airway epithelia. These findings suggest that commercial multi-walled CNT interfere with the formation and the maintenance of tight junctional complexes in airway epithelial cells. PMID- 18403141 TI - Carboxylated high amylose starch as pharmaceutical excipients. Structural insights and formulation of pancreatic enzymes. AB - Carboxymethyl high amylose starch (CM-HAS) and succinate high amylose starch (S HAS) were proposed as pharmaceutical excipients for oral drug delivery, providing a significant gastroprotection to dosage forms of pancreatic enzymes (alpha amylase, lipase and trypsin) compared to unprotected enzymes. In acidic medium, carboxylic groups are protonated (at least in tablet surface) ensuring local buffering properties and giving a compact shape of the tablets. The enzymes were formulated individually or in association as three enzymes formulation. After the first hour of incubation (over a 2h experiment) in simulated gastric fluid (SGF), the three pancreatic enzymes retained an overall (average of the three enzymes) activity of 72% when formulated as tablets with CM-HAS excipient and 77% when formulated with S-HAS excipient. Furthermore, after incubation in SGF, the delivery of 75% of the total remaining enzymatic activity in the simulated intestinal fluid (SIF) taken 180 and 170 min for CM-HAS and S-HAS, respectively. Both formulations with carboxylated starch as excipient have a high loading capacity (up to 70-80% enzymes), which is of interest for pancreatic enzymes replacement therapy of pancreatitis. An advantage of these formulations is that gastroprotection is afforded by the carboxylated matrices (carboxylic groups), without enteric coating. PMID- 18403142 TI - Lyotropic liquid crystal preconcentrates for the treatment of periodontal disease. AB - The aim of our study was to develop water-free lyotropic liquid crystalline preconcentrates, which consist of oils and surfactants with good physiological tolerance and spontaneously form lyotropic liquid crystalline phase in aqueous environment. In this way these preconcentrates having low viscosity can be injected into the periodontal pocket, where they are transformed into highly viscous liquid crystalline phase, so that the preparation is prevented from flowing out of the pocket due to its great viscosity, while drug release is controlled by the liquid crystalline texture. In order to follow the structure alteration upon water absorption polarization microscopical and rheological examinations were performed. The water absorption mechanism of the samples was examined by the Enslin-method. Metronidazole-benzoate was used as active agent the release of which was characterized via in vitro investigations performed by means of modified Kirby-Bauer disk diffusion method. On the grounds of the results it can be stated that the 4:1 mixture of the investigated surfactants (Cremophor EL, Cremophor RH40) and oil (Miglyol 810) formed lyotopic liquid crystalline phases upon water addition. Polarization microscopic examinations showed that samples with 10-40% water content possessed anisotropic properties. On the basis of water absorption, rheological and drug release studies it can be concluded that the amount of absorbed water and stiffness of lyotropic structure influenced by the chemical entity of the surfactant exerted major effect on the drug release. PMID- 18403143 TI - Famotidine polymorphic transformation in the grinding process significantly depends on environmental humidity or water content. AB - The effect of environmental humidity and additional water added on the polymorphic change of famotidine in the process of grinding was investigated. The famotidine form B powder with or without additional amount of water added was respectively ground for 30 min in an oscillatory ball mill under 25+/-2 degrees C and three relative humidities (RH) (50+/-5%, 75+/-5% or 95+/-5% RH). Each ground sample was periodically isolated for analytical determinations by using differential scanning calorimetry (DSC), thermogravimetric (TG) analysis and Fourier transform infrared (FT-IR) microspectroscopy. The results indicate that the higher environmental humidity might induce and promote the polymorphic transformation of famotidine from form B to form A in the process of grinding. Moreover, the more the amount of water externally added the easier the polymorphic transformation of famotidine from form B to form A obtained. In addition, the more grinding time spent the more formation of form A obtained. This study disavowed the results of other studies in which no polymorphic change of famotidine even by grinding. The apparent evidence shows that the solid-state polymorphic transformation of famotidine from form B to form A in the grinding process significantly depended on the relative humidity of atmosphere and the additional amount of water added. PMID- 18403144 TI - Evaluation of protective effects of Chi-Zhi-Huang decoction on Phase I drug metabolism of liver injured rats by cocktail probe drugs. AB - AIM OF THE STUDY: Chi-Zhi-Huang decoction (PGR) is one of the traditional Chinese medicine (TCM) preparations with unique effect on withdrawing jaundice and has been used to treat icteric patients in China for many years. In this research, we aim at to evaluate the potential activity of PGR in restoring hepatic drug metabolism in a damaged liver. MATERIALS AND METHODS: A cocktail approach with caffeine (10mg/kg), dapsone (10mg/kg) and chlorzoxazone (20mg/kg) respectively as probe drug of cytochrome P450 (CYP) isoform of CYP 1A2, 3A4 and 2E1 was used to evaluate its possible effects on Phase I oxidative metabolism. Pretreated with three dosages of PGR water extract (0.75, 1.5 and 3g/kg, po) for 5 days, male Wistar rats (220-240 g) were intoxicated by phenylisothiocyanate (PITC, 100mg/kg, po) 24h before probes intravenous injection. The pharmacokinetics of the probes in the blood was determined simultaneously by HPLC, and their non-compartmental parameters were used to evaluate the metabolic difference among the groups. Moreover, the levels of liver enzymes (ALT, AST, ALP) and bilirubins were also measured for insight of liver function. RESULTS: The findings in this study suggest that PGR induces CYP 3A4, does not have much effect on CYP 2E1, and inhibits CYP 1A2 at high dosage. CONCLUSION: The current pharmacokinetic approach allowed the protective effects of PGR on oxidative drug metabolism in damaged liver to be systemically examined and will certainly help in the explanation of synergistic effect of the composites formula. PMID- 18403145 TI - Anti-hypertensive effects of the methanol/methylene chloride stem bark extract of Mammea africana in l-NAME-induced hypertensive rats. AB - AIM OF THE STUDY: The methanol/methylene chloride (CH(3)OH/CH(2)Cl(2)) extract from the stem bark of Mammea africana was showed to possess vasodilating effect in the presence and the absence of N(omega)-nitro-l-arginine methyl ester (l NAME). The present study was designed to evaluate the effects of the methanol/methylene chloride from the stem bark of Mammea africana. MATERIALS AND METHODS: The extract (200 mg/(kg day)) was administered orally in rats treated concurrently with l-NAME (40 mg/(kg day)). l-Arginine (100 mg/(kg day)) and captopril (20 mg/(kg day))were used as positive controls. Bodyweight, systolic arterial blood pressure and heart rate were measured weekly throughout the experiment period (28 days). At the end of treatment, animals were killed and the cardiac mass index evaluated. The aorta was used to evaluate the endothelium dependant relaxation to carbachol. The aorta contraction induced by noradrenalin was also examined and expressed as a percentage of that induced by KCl. RESULTS: The extract neither affected the body weight nor the heart rate. The extract as captopril completely prevented the development of arterial hypertension. Both the substances failed to restore the endothelium-dependent vascular relaxation and increased the vascular contraction to norepinephrine in relation to KCl contraction. They also significantly reduced the left ventricular hypertrophy induced by l-NAME. CONCLUSION: These findings are in agreement with the traditional use of Mammea africana in the treatment of arterial hypertension and indicate that it may have a beneficial effect in patients with NO deficiency but will be unable to improve their endothelium-dependent vasorelaxation. PMID- 18403146 TI - [Thoracic chest wall fistula formation]. AB - Thoracic actinomycosis is a suppurative infection which can be difficult to diagnose as its presentation may mimic cancer or tuberculosis. We report a new case of thoracic actinomycosis in a 35-year-old man who presented with thoracic symptoms associated to a productive parietal fistula. Imaging exploration revealed an opacity of the right ventroapical segment with parietal infiltration. A bilobectomy and a parietectomy were performed. The anatomopathologic diagnosis actinomycosis was confirmed. The patient was first put on a treatment of azathioprine 1g daily during two weeks, then switched to a combination with Vibramycin 100 mg twice a day during 17 months, The evolution was marked by the persistence of productive fistulae, which were treated surgically, and resistance to the initial treatment leading to a switch to Augmentin 3 g daily during 25 days. The patient experienced clinical improvement with a follow up of 18 months than was lost to follow-up. PMID- 18403147 TI - Clinical, sonographic and manometric characteristics and impact on quality of life of fecal incontinence in 92 men referred for endoanal ultrasonography. AB - Anal incontinence (AI) is a frequent symptom with considerable impact on quality of life. The aim of this study was to describe the clinical, sonographic and manometric characteristics of a male population with AI. MATERIALS AND METHODS: Endoanal ultrasonography (EAU) was performed in 92 men referred for exploration of AI. Anal incontinence severity was evaluated by the Jorge and Wexner score (JW). The gastrointestinal quality-of-life index (GIQLI) was determined in 57% of patients. Anorectal manometry was performed in 62.6% of patients. RESULTS: The average JW score was 11+/-1. Anal incontinence had considerable impact on quality of life: average GIQLI=81+/-4. Seventeen patients presented an anal sphincter defect on EAU, 16 of whom had a history of coloproctological surgery. Prior surgery was significantly more common among patients who had a defect on ultrasonography; manometry showed significantly lower resting anal pressure. CONCLUSION: Our study confirms the severity of AI in a male population and its impact on quality of life. It also highlights the high prevalence of anal sphincter defects in patients with a history of anal surgery. PMID- 18403148 TI - [Doppler-guided hemorrhoidal artery ligation: the new deal of surgical treatment of hemorrhoids]. PMID- 18403149 TI - [Verner-Morrison syndrome revealing a ganglioneuroblastoma in an adult]. AB - Most vasoactive intestinal peptide (VIP)-producing tumours are from epithelial origin. Tumours derived from the sympathetic nervous system can produce VIP as well. We report here the case of a Verner-Morrison syndrome in a 40-year-old woman revealing a metastatic ganglioneuroblastoma. The diarrhea resolved after the resection of primary tumour and liver metastases. Neuroblastic tumours occur extremely rarely in adults. Thus, the management of these tumours is poorly defined in adults. PMID- 18403150 TI - Phenotypic expression of ferroportin disease in a family with the N144H mutation. AB - Ferroportin is a putative transmembrane channel involved in the exit of iron out of the enterocytes, the macrophages and the hepatocytes. Mutations in the human gene coding ferroportin have been linked to an unusual form of iron overload, now referred to as "hemochromatosis type IV" or "ferroportin disease" characterized by a prevalent iron overload of macrophages and liver Kupffer cells. We report four patients from a same family with ferroportin disease associated with the N144H mutation. We show that in this family the mutation which is fully penetrant, may act through an increased iron export from macrophages as suggested by the unexpected absence of iron overload in the spleen and bone marrow detected by magnetic resonance imaging, that it co-segregates with a phenotype close to the classical form of HFE-associated hemochromatosis and was associated, in the oldest patient, with the development of hepatocellular carcinoma in a non cirrhotic liver. Our findings illustrate the existence of a genotype-phenotype relationship in "ferroportin disease", suggest that MRI may be useful in determining this phenotype and show that hepatocellular carcinoma may occur in these patients even without cirrhosis. This observation justifies careful follow up of this subgroup of patients. PMID- 18403151 TI - [New rules for distributing liver grafts]. PMID- 18403152 TI - Fortuitous discovery of common bile duct stones: results of a conservative strategy. AB - OBJECTIVE: The incidence of fortuitously discovered stones in the common bile duct is about 5%. The purpose of this study was to determine the rate of spontaneous clearance of asymptomatic stones in the common bile duct discovered fortuitously during cholecystectomy. PATIENTS AND METHODS: Intraoperative cholangiography was performed in all patients undergoing cholecystectomy for symptomatic gallbladder stones. If a filling defect of the common bile duct was discovered, a transcystic drain was inserted. Surgical or endoscopic extraction was not proposed initially. A control cholangiogram was performed on the second postoperative day then during the sixth postoperative week. If a stone persisted at the sixth week, endoscopic extraction was undertaken. RESULTS: Cholecystectomy was performed in 124 patients with symptomatic gallstones and no signs predictive of stones in the common bile duct. A stone was found fortuitously in the common bile duct in 12 patients. The control cholangiogram was normal in two of these patients on day two (16.7%) and in six others (50%) at the six-week control. All 12 patients remained free of symptoms suggesting the presence of a stone in the common duct. Presence of the drain had no impact on quality-of-life. Endoscopic extraction was finally performed for four patients (33.3%) to remove a stone from the common bile duct. CONCLUSION: Early surgical or endoscopic extraction of stones in the common bile duct should not be undertaken systematically in asymptomatic patients. Spontaneous asymptomatic clearance of the common bile duct is observed in about half of patients. PMID- 18403153 TI - Long-term results of liver transplantation for Wilson's disease. AB - Wilson's disease is a hereditary defect in hepatic copper metabolism, causing hepatic, neurological and/or psychiatric manifestations. For patients with severe disease, liver transplantation is the treatment of choice. The aim of this study was to report the long-term outcome of patients who underwent liver transplantation for Wilson's disease. PATIENTS AND METHODS: Thirteen patients with Wilson's disease, transplanted in Lyon France between January 1987 and May 2006, were including in this study: eight women and five men, aged eight to 53 years (median 20 years, seven children and six adults). The diagnosis of Wilson's disease was established before liver transplantation. RESULTS: The indication for liver transplantation was chronic (69%) or fulminant liver failure (31%). The median follow-up after liver transplantation was 10 years with 100% patient survival. Copper metabolism returned to normal in all patients. None of the patients with exclusive liver disease required chelation treatment after liver transplantation and none developed neurological symptoms of Wilson's disease. CONCLUSION: Liver transplantation totally reverses the abnormalities of copper metabolism and subsequent hepatic failure, but the course of neurological symptoms remains unpredictable. Long-term patient survival can be excellent without occurrence of neurological complications. PMID- 18403154 TI - [A royal fistula in unexpected consequences]. PMID- 18403155 TI - [Right liver agenesis with ectopic gallbladder and bile duct calculi]. PMID- 18403156 TI - Sonographic preoperative assessment of liver volume before major liver resection. AB - OBJECTIVE: The use of ultrasonography is widespread for both the diagnosis and treatment of liver tumors. However, the measurement of liver volume by ultrasonography is not commonly done. We report an original method of liver volumetry using ultrasonography and an investigation into the usefulness of ultrasonography in this context. METHODS: The data for 50 patients undergoing various types of major hepatectomy were collected. We preoperatively measured liver volume using ultrasonography, dividing the liver into three main compartments according to precise anatomical landmarks, and then made comparisons with the volume of the actual specimen after hepatectomy, for all of the study participants. RESULTS: Total volume correlation between the two groups was good (r = 0.916, P < 0.001). However, the correlation was weaker in cases of right hepatectomy compared with other types of hepatectomy. CONCLUSION: This study demonstrates the possibility of doing liver volumetry using an ultrasound device. Further investigation to establish the reliability of this easily available and noninvasive approach is needed. PMID- 18403157 TI - Pseudotumors of the shoulder invited review. AB - This paper discusses the main types of MRI pseudotumors in and around the shoulder region. Some unusual types of pseudotumor will also be mentioned. Suggestions on how to improve awareness and diagnosis are also given. PMID- 18403158 TI - Atomic absorption spectrometric determination of the iridium content in tumor cells exposed to an iridium metallodrug. AB - An electrothermal atomic absorption spectrometric method to quantify the iridium content of HT-29 colon carcinoma cells exposed to iridium metallodrugs was developed. Optimisation of the procedure involved the evaluation of pyrolysis and atomisation conditions (optimal values were 1400 degrees C for pyrolysis and 2400 degrees C for atomisation) and the addition of appropriate additives. The presence of cellular components as well as the addition of nitric acid and hydrochloric acid led to enhanced absorption signals and suggested the use of matrix matched calibration. The described method allows the measurement of iridium in cell suspensions in the low microg/L range (linear dynamic range: 10 450 microg/L) with a detection limit of 11.2 microg/L. The applicability of the method was tested by means of a novel iridium metallodrug. First results on the complex [IrCl3(DMSO)(phen)] indicated a low cellular uptake (21.2 microM at incubation with 100 microM) of this iridium species in HT-29 cells compared to other metal containing antitumor drugs. PMID- 18403159 TI - Fast and simple method for assay of ciclopirox olamine by micellar electrokinetic capillary chromatography. AB - A rapid, simple and specific method has been developed and validated for the assay of ciclopirox olamine in pharmaceutical formulations using micellar electrokinetic capillary chromatography (MEKC). The key factors, including pH, buffer concentration and buffer additive, sodium dodecyl sulfate (SDS) concentration, applied voltage and injection time have been systematically investigated in a fused silica capillary (i.d. 50 microm, total length 45 cm and effective length 38 cm) with UV detection at 298 nm. Optimized conditions have been established on the basis of the experimental results. The buffer contains 200 mM borate, 20 mM SDS and 2 mg mL(-1) EDTA at pH 8.0 and the applied voltage is 20 kV with hydrodynamics sample injection (15 cm high for 5s). The method has been validated with respect to its specificity, linearity, limits of detection, and quantification, precision and accuracy. The total analysis time was less than 10 min with good peak shape for ciclopirox olamine, which eluted at 3.6 min. Degradation of the ciclopirox olamine was forced using different conditions. These were using hydrogen peroxide, acidic and basic conditions, heat and light. The degradation products so produced showed no interference with ciclopirox olamine. A linear standard curve was established over the concentration range 31.3-2.00 x 10(3) microg mL(-1) of ciclopirox olamine in running buffer with a correlation coefficient (r) of 0.9999. The limits of quantification and detection were 31.3 and 9.36 microg mL(-1), respectively. The proposed method has been successfully used for the quantitative determination of ciclopirox olamine in pharmaceutical suppository and cream formulations. PMID- 18403160 TI - Development and validation of dissolution test for lopinavir, a poorly water soluble drug, in soft gel capsules, based on in vivo data. AB - The objective of the present study was to develop and validate a dissolution test for lopinavir soft gel capsules (Kaletra), using a simulated absorption profile based on in vivo data. Different conditions such as surfactant concentration, apparatus and rotation speed were evaluated. In vivo release profiles were obtained from the literature. The fraction (and percentage) of dose absorbed (FA) was calculated by using Wagner-Nelson method. The best in vitro dissolution profile was obtained using Apparatus 2 (paddle) at 25 rpm, 1000 ml of medium with 2.3% of sodium lauryl sulfate and pH 6.0. Under these conditions a level-A in vitro-in vivo correlation (IVIVC) was obtained (r = 0.997). The in vitro dissolution samples were analyzed using a HPLC method and the validation was performed according to USP protocol. The method showed accuracy, precision, linearity and specificity within the acceptable range. Both the HPLC method and the in vitro dissolution method were validated and could be used to evaluate the release profile of lopinavir soft gel capsules. PMID- 18403161 TI - Validated stability-indicating HPLC method for the determination of dimethyl-4,4' dimethoxy-5,6,5',6'-dimethylene dioxybiphenyl-2,2'-dicarboxylate (DDB) and its degradation products. AB - High-performance liquid chromatographic method was developed for the quantitative determination of dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylene dioxybiphenyl-2,2' dicarboxylate (DDB) and its degradation products. Forced degradation studies were performed on bulk sample of DDB using acid (1N hydrochloric acid), alkaline (0.1N sodium hydroxide), oxidation (0.33% hydrogen peroxide), heat (70 degrees C) and photolytic degradation. The chromatographic method was fine tuned using the samples generated from forced degradation studies. Good resolution between the peaks corresponds to degradation products and the analyte was achieved on 5 microm ODS column (Luna, Phenomenex, USA). The mobile phase consists of a mixture of acetonitrile and water (60:40, v/v). Quantitation was achieved with UV detection at 235 nm based on peak area. The proposed HPLC method was utilized to investigate the kinetics of acidic, alkaline and oxidative degradation processes of DDB at different temperatures and the apparent pseudo first-order rate constant, half-life and activation energy were calculated. The pH-rate profiles of degradation of DDB in Britton-Robinson buffer solutions within the pH range 2 11 were studied. The developed method was validated with respect to linearity, accuracy, precision, robustness and forced degradation studies prove the stability-indicating power of the method. PMID- 18403162 TI - Determination of glycine and threonine in topical dermatological preparations. AB - In the present study, a single HPLC method was developed for the determination of glycine and threonine in cicatrizants. Two different preparations of a cream and an ointment, and the corresponding bandages, onto which the formulations were applied, were studied. The method involved matrix solubilisation with dichloromethane, liquid-liquid isolation of gly and thr with aqueous 1N NaOH, and derivatization with phenylisothiocyanate. Reversed-phase HPLC separation was carried out by gradient elution with 20mM aqueous NaClO4 and acetonitrile (from 90% to 30% aqueous NaClO4 in 10 min) on a LiChrospher 100 RP-18 cartridge (125 mm x 4.6 mm). Analytes were determined with a UV detector set at 245 nm. Quantitation was accomplished by internal standardization with methionine. Linearity was studied in the range 60-120% of the concentrations expected for gly and thr (viz. for gly from 200 to 400 microgml(-1), and for thr from 100 to 200 microgml(-1)). In reference aqueous samples, linear correlation (r) was better than 0.99 for gly and thr, while in spiked matrix samples r ranged from 0.97 to 0.98. Recoveries were in the 95-105% interval, and precision (CV%, N=6) was better than 5% for both analytes either in cream, ointment or bandages. The method was successfully used for the quality control of topical dermatological preparations. PMID- 18403163 TI - Is the intra-aortic balloon pump a method of brain protection during cardiogenic shock after drug intoxication? AB - Cardiovascular medications are ubiquitous and are frequently implicated in accidental or intentional overdose. It is common that combined use of these drugs may lead to hypotension and even shock, followed by metabolic derangements. We report a case in which an intra-aortic balloon pump (IABP) was used in the management of self-poisoning with verapamil, amlodipine, metoprolol, and ibuprofen. In presenting this case of combined massive drug ingestion, we outline early strategy in the Emergency Department and some alternative treatment options. Beyond pharmacological and conservative procedures, we implemented an invasive approach that included temporary pacing, mechanical ventilation, and intra-aortic balloon counterpulsation (IABP). Such intense treatment was necessary due to the critical state of the patient. In our opinion, the use of the IABP contributed to the final recovery of our adolescent patient. Combined mechanical and pharmacological treatment may protect from multi-organ insufficiency, including permanent central nervous system injury. It is hoped that reporting our experience will raise awareness of alternative treatment options for ingestions of cardiovascular medications. PMID- 18403164 TI - Association of injury visits in children and child maltreatment reports. AB - Injuries are a leading cause of childhood morbidity and are also common manifestations of child maltreatment, especially among young children. In an effort to determine whether injury-related Emergency Department (ED) visits among children aged 0 to 4 years were associated with child maltreatment reports, we identified all children with at least one injury-related ED visit in Missouri during 2000. Data on these injured children were linked to Missouri Child Protective Services (CPS) child abuse and neglect reports for 2000 and 2001. There were 50,068 children with at least one injury-related ED visit. Using children with one injury-related ED visit as the reference category, we calculated the relative risk of having a CPS report (or a substantiated report) for children with two, three, and four or more ED visits before a CPS report (or substantiated report). Compared to children with one visit, children with two visits were more likely to have a CPS report (relative risk [RR] 1.9; 95% confidence interval [CI] 1.8-2.0) and a substantiated report (RR 2.5; 95% CI 2.1 2.9). For children with four or more visits, the relative risk of a report and substantiated report was 3.8 (95% CI 3.0-4.7) and 4.7 (95% CI 2.4-9.2), respectively. Children with two or more injury-related ED visits in 1 year are more likely to be reported for child maltreatment and to have a substantiated report. PMID- 18403166 TI - Learning from our history. PMID- 18403165 TI - Pattern of orlistat exposures in children aged 5 years or less. AB - On February 7, 2007, orlistat became the first weight-loss drug approved by the United States Food and Drug Administration for over-the-counter sales. However, information on exposures among young children is limited. The objective of this study was to describe the pattern of orlistat exposures among young children reported to poison control centers. The pattern of all exposures to orlistat alone among patients < or = 5 years old reported to six poison control centers during 1999-2005 was identified with respect to various factors. There were 107 cases. The average age was 21.4 months. There were 55 males, 51 females, and 1 unknown. The dose was identified for 76 cases. The mean dose was 155 mg. Patients were managed on site in 88% of the cases, were already at a health care facility in 8%, and were referred to a health care facility in 5%. Of the 45 patients with a known medical outcome, the outcome was no effect for 91% and minor effect for 9% of the patients. Of the 92 cases reported during 2000-2005, the listed adverse clinical effects were diarrhea (n = 4) and vomiting (n = 1), and the listed treatments were decontamination by dilution (n = 62), food (n = 8), activated charcoal (n = 5), other emetic (n = 2), cathartic (n = 1), and ipecac (n = 1). Orlistat exposures among young children involving small doses encountered by poison control centers can usually be managed on site through decontamination, and have favorable outcomes with few adverse clinical effects, mainly gastrointestinal in nature. PMID- 18403167 TI - Ophthalmic exposure to crotalid venom. AB - Crotalid venom exposure to the eye is uncommon. A 40-year-old woman sustained an accidental mucus membrane exposure of rattlesnake venom to her face and right eye. She was successfully treated with irrigation, topical antibiotics, and intravenous crotalid antivenin. PMID- 18403168 TI - Diagnosis of Lemierre syndrome by bedside emergency department ultrasound. AB - Oropharyngeal infections such as pharyngitis and odontogenic abscess are routinely encountered in emergency and primary care medical practice. Lemierre syndrome is a rare but serious complication of such infections. This syndrome is characterized by penetration of the primary infection into the lateral pharyngeal space, suppurative thrombophlebitis of the internal jugular vein, and metastatic infections resulting from septic emboli. A combination of clinical suspicion, microbiologic identification of the causative organism, and diagnostic imaging may be required to make the diagnosis. We present a case of Lemierre syndrome that was rapidly diagnosed in the Emergency Department with bedside ultrasound of the internal jugular vein. This case suggests that bedside ultrasound, performed before other radiologic imaging, may lead to earlier diagnosis and treatment of this syndrome, which historically has been associated with significant morbidity and mortality. PMID- 18403169 TI - Ovarian hyperstimulation syndrome: a potentially fatal complication of early pregnancy. AB - We present the case of a patient who presented to the Emergency Department (ED) 48 h after successful in vitro fertilization with abdominal pain, hypotension, and free fluid on an ED-focused abdominal sonogram for trauma study. This presentation is typical of Ovarian Hyperstimulation Syndrome (OHSS), a diagnosis that may be unfamiliar to many Emergency Physicians. With the increasing frequency of in vitro fertilization procedures, this disease process is becoming more common. Numerous complications can occur with OHSS, including third-space fluid accumulation, hemoconcentration, renal failure, and thromboembolic phenomena. Vigilance is required as these patients are at increased risk of ovarian torsion, ovarian rupture with internal hemorrhage, ectopic pregnancy, and infection. This case report provides an overview of clinical features and emergent management of OHSS. PMID- 18403170 TI - Emergency department diagnosis of pediatric hip effusion and guided arthrocentesis using point-of-care ultrasound. AB - Children with complaints of hip pain, a painful limp, or refusal to weight bear commonly present to the Emergency Department (ED). The ability to use point-of care ultrasound in the ED to diagnose a hip joint effusion and to guide arthrocentesis can be helpful to facilitate diagnosis and management of these children. The capsular-synovial thickness of the hip is measured from the anterior bony cortical surface to the posterior surface of the iliopsoas muscle at the concavity of the femoral neck. A capsular-synovial thickness>5 mm, or >2 mm difference compared to the asymptomatic contralateral hip are the described sonographic criteria for hip joint effusion in children. We report on the use of point-of-care ultrasound to diagnose hip effusion and to guide arthrocentesis in a series of pediatric patients presenting with hip pain to the ED. PMID- 18403171 TI - Third-degree AV block from extended-release diltiazem ingestion in a nine-month old. AB - Calcium channel blocker (CCB) overdose is associated with dysrhythmias and atrioventricular (AV) block, however, experience with infant CCB overdose is limited. A 9-month-old girl was found playing with tablets of extended-release diltiazem 120 mg. The patient had two episodes of emesis, which contained pill fragments, and was brought to the Emergency Department (ED) 4.5 h after being found. Vital signs were: rectal temperature 37.1 degrees C, pulse 87 beats/min, respiratory rate 30-40 breaths/min, blood pressure 72/48 mm Hg, and oxygen saturation (SpO(2)) 99% on room air. Otherwise, the patient was well-appearing, with normal skin color and examination. The electrocardiogram revealed third degree atrioventricular block with a ventricular rate of 90 beats/min, QRS 68 ms, and QTc 411 ms. Atropine 0.1 mg i.v. was given, which increased the heart rate to 100-110 beats/min. Calcium gluconate 500 mg was also given intravenously. Laboratory evaluation revealed bicarbonate 17 mEq/L, anion gap 16, and glucose 129 mg/dL. On hospital day 1, the patient was noted to have a junctional rhythm with a rate of 90-100, and systolic blood pressure of 80-90 mm Hg. No additional medications were given. Early on day 2, the patient converted spontaneously to a normal sinus rhythm and was discharged approximately 42 h after presentation to the ED. In addition to bradycardia and hypotension, this 9-month-old patient manifested third-degree AV block after ingesting extended-release diltiazem. PMID- 18403172 TI - Special considerations in hazardous materials burns. AB - Those practicing Emergency Medicine are frequently faced with a patient presenting with a chemical burn. Most dermal chemical burns are minor and do not require specialized treatment. Occasionally, however, the clinician may be in the position of responding to a chemical burn in which standard therapy of irrigation and good wound care may not be sufficient or, at worst, contraindicated. Several burn conditions will be reviewed, some of those requiring only specific decontamination techniques, as in hot tar, others posing special hazards to clinicians, as in elemental metals, and finally, examples are given of hazardous materials requiring attention to systemic effects, as in hydrofluoric acid. PMID- 18403173 TI - Immunity protection and delayed transfer of patient care by EMS providers: a medical-legal analysis. AB - In emergency medical service (EMS) systems, the transfer of patient care to persons at the receiving facility is delayed when EMS providers stop en route to the facility, or postpone entering after arrival, to perform tasks. When these tasks are prolonged and inessential, the delay in transferring care is judged to be inappropriate. When transfer of care is inappropriately delayed, EMS providers, supervisors, and medical directors may lose the immunity provided by their state's EMS Act. This article analyzes the legal issues surrounding inappropriate delays in transfer of care by EMS providers. Loss of statutory immunity may occur for reasons of public policy, as reflected in case law and under the reasonableness standard. Without immunity, persons involved in the transfer of patients to receiving facilities may be subject to liability under ordinary rather than gross negligence standards. PMID- 18403174 TI - Brugada syndrome presenting as an "acute myocardial infarction". AB - Brugada syndrome is believed to be the cause of up to 50% of sudden cardiac death (SCD) cases due to ventricular dysrhythmias in young healthy individuals with no structural heart disease. This syndrome was first reported in 1992 and is rarely seen in the Emergency Department (ED). Given the life-threatening nature of Brugada syndrome, we hope to increase awareness in Emergency Medicine practitioners. We report the case of a 22-year-old man who was referred to the ED with a history of intermittent palpitations, near-syncope and electrocardiogram findings of RSR' and ST elevation in V1-V2 characteristic of Brugada syndrome. It is crucial that emergency physicians search for this diagnosis, as an implantable cardioverter-defibrillator is the only recognized life-saving intervention, and the risk of SCD is high if the diagnosis is missed. PMID- 18403175 TI - Knowledge integration: conceptualizing communications in cancer control systems. AB - OBJECTIVE: This paper was prepared by the National Cancer Institute of Canada (NCIC) Working Group on Translational Research and Knowledge Transfer. The goal was to nurture common ground upon which to build a platform for translating what we know about cancer into what we do in practice and policy. METHODS: Methods included expert panels, literature review, and concept mapping, to develop a framework that built on earlier cancer control conceptualizations of communications that have guided researchers and end users. RESULTS: The concept of 'knowledge integration' is used to describe the resulting refinement and the nature of evidence necessary for decision-making to at the systems level. Current evidence for knowledge integration in cancer control is presented across the levels of individual, organizational and systems level interventions and across basic, clinical and population science knowledge bases. CONCLUSION: A systems oriented approach to integrating evidence into action assists organizations to conduct research and policy and practice. PRACTICE IMPLICATIONS: Practitioners can use this framework to understand the challenges of implementing and evaluating cancer control strategies. PMID- 18403176 TI - Roles of IGF-I and the estrogen, androgen and IGF-I receptors in estradiol-17beta and trenbolone acetate-stimulated proliferation of cultured bovine satellite cells. AB - Although numerous studies have shown that both androgenic and estrogenic steroids increase rate and efficiency of muscle growth in steers, there is little consensus as to their mechanism of action. A combined estradiol 17beta (E2)/trenbolone acetate (TBA) implant causes a significant increase in muscle IGF I mRNA and both E2 and TBA stimulate a significant increase in IGF-I mRNA level in bovine satellite cell (BSC) cultures in media containing 10% fetal bovine serum (FBS). Consequently, increased IGF-I expression may play a role in anabolic steroid-enhanced muscle growth. However, even though treatment of cultured BSC with E2 or TBA in media containing 1% IGFBP-3-free swine serum (SS) results in increased proliferation there is no effect on IGF-I mRNA expression, suggesting that increased IGF-I expression may not be responsible for anabolic-steroid enhanced BSC proliferation. To further examine the role of estrogen, androgen and IGF-I receptors and their respective ligands in E2- and TBA-stimulated BSC proliferation, we assessed the effects of specific inhibitors on E2- or TBA stimulated proliferation of BSC. Both ICI 182 780 (an estrogen receptor blocker) and flutamide (an inhibitor of androgen receptor) suppressed (p<0.05) E2- and TBA stimulated BSC proliferation, respectively. JB1 (a competitive inhibitor of IGF-I binding to type I IGF receptor) reduced (p<0.05) both E2- and TBA-stimulated proliferation in BSC cultures. Both the Raf-1/MAPK kinase (MEK)1/2/ERK1/2, and the phosphatidylinositol 3-kinase (PI3K)/Akt pathways play significant roles in the actions of IGF-I on proliferation and differentiation of myogenic cells. PD98059, an inhibitor of the MAPK pathway, and wortmannin, an inhibitor of the PI3K pathway, both suppressed (p<0.05) E2- and TBA-stimulated proliferation of cultured BSC. Our data suggest that IGF-I plays a role in E2- and TBA-stimulated proliferation of cultured BSC even in the absence of increased IGF-I expression. PMID- 18403177 TI - [Massive right auricular and ventricular thrombosis after Kaskadil administration in a patient with an overdose of antivitamin K]. PMID- 18403178 TI - Deformable brain atlas validation of the location of subthalamic nucleus using T1 weighted MR images of patients operated on for Parkinson's. AB - Parkinson's disease is a degenerative disease of the central nervous system. One of the most effective treatments is deep brain stimulation. This technique requires the localization of an objective structure: the subthalamic nucleus. Unfortunately this structure is difficult to locate. In this work the creation of a deformable brain atlas that enables the identification of the subthalamic nucleus in T1-weighted magnetic resonance imaging (MRI) in an automatic, precise and fast way is presented. The system has been validated using data from 10 patients (20 nucleus) operated on for Parkinson's. Our system offers better results using a Wendland function with an error of 1.8853+/-0.9959mm. PMID- 18403179 TI - SketchSnakes: sketch-line initialized Snakes for efficient interactive medical image segmentation. AB - We present an intuitive, fast and accurate 2D interactive segmentation method that combines a general subdivision-curve Snake possessing powerful editing capabilities, with a novel sketch-line user initialization process, and a pen input device. Using the pen (or a mouse), the Snake is quickly and precisely initialized with a few quick sketch lines drawn across the width of the target object. The smooth contour constructed using these lines is extremely close to the position and shape of the object boundary. This makes the Snake's task of snapping to the object boundary much simpler and hence more likely to succeed in noisy images with minimal user editing. We apply our Snake to the segmentation of several 2D medical images to demonstrate it's efficiency, accuracy and robustness. We also compare SketchSnakes to Adobe Photoshop's Magnetic Lasso (Adobe Systems Inc., Adobe Photoshop User Guide, 2002) as well as a recent graph cut based image cutout tool known as Snap (Digital Film Tools LLC, Snap User Guide, 2007) in order to highlight SketchSnakes effectiveness. PMID- 18403181 TI - Is oral nutritional therapy effective for the treatment of chylothorax? A case report. AB - We report a case of a female patient who underwent corrective aortic coarctation surgery that progressed to chylothorax on the fifth postoperative day. Because the patient was clinically stable and had a functioning digestive tract, the nutritional team decided to treat her by oral nutritional support with a low lipid diet, rich in medium-chain triacylglycerols. After 20 d, the patient returned to her habitual home diet and did not develop pleural spilling, showing full healing of the thoracic duct. PMID- 18403180 TI - Zfra is an inhibitor of Bcl-2 expression and cytochrome c release from the mitochondria. AB - Zfra is a small size 31-amino-acid C2H2 zinc finger-like protein, which is known to interact with c-Jun N-terminal kinase 1 (JNK1), WW domain-containing oxidoreductase (WWOX, FOR or WOX1), TNF receptor-associated death domain protein (TRADD) and nuclear factor kappaB (NF-kappaB) during stress response. Here, we show that Zfra became phosphorylated at Ser8 (as determined by specific antibody) and translocated to the mitochondria in response to inducers of mitochondrial permeability transition (MPT) (e.g. staurosporine and betulinic acid). Overexpressed Zfra induced cell death. This event is associated, in part, with increased dissipation of mitochondrial membrane potential (MMP) and increased chromosomal DNA fragmentation. Intriguingly, Zfra significantly downregulated Bcl 2 and yet blocked cytochrome c release from the mitochondria. Overexpression of an S8G-Zfra mutant (Ser8 to Gly8 alteration) could not induce cell death, probably due to its failure of translocating to the mitochondria and causing MMP dissipation. Over-expressed proapoptotic WOX1 induced cytochrome c release from the mitochondria. Zfra bound and blocked the effect of WOX1. Taken together, Ser8 is essential for overexpressed Zfra to exert cell death via the mitochondrial pathway. Zfra downregulates Bcl-2 and induces MMP dissipation but causes no cytochrome c release, indicating a novel death pathway from the mitochondria. PMID- 18403182 TI - Safety and efficacy of a graduated intravenous insulin infusion protocol in critically ill trauma patients receiving specialized nutritional support. AB - OBJECTIVE: The intent of this study was to evaluate the safety and efficacy of our protocol for providing continuous intravenous regular human insulin (RHI) infusion to hyperglycemic critically ill trauma patients receiving specialized nutritional support. METHODS: Capillary blood glucose (BG) concentrations were determined every 1-2 h. Glucose control was defined as a BG concentration in the target range of 70-149 mg/dL (3.9-8.3 mmol/L). Data were recorded for 1 d before the RHI infusion and for a maximum of 8 d thereafter while receiving the RHI infusion. RESULTS: Forty adult critically ill trauma patients received 102 +/- 62 units of RHI daily for 10 +/- 6 d. BG control was achieved within 5 +/- 3 h. BG decreased from 194 +/- 55 mg/dL (10.8 +/- 3.1 mmol/L) to 134 +/- 19 mg/dL (7.4 +/ 1.1 mmol/L) after 1 d of RHI infusion (P < 0.001). Average daily BG ranged from 119 to 124 mg/dL and the target range was maintained for 19.6 +/- 4.7 h/d. None of the patients experienced severe hypoglycemia (<40 mg/dL); 14 patients had asymptomatic hypoglycemia (<60 mg/dL or <3.3 mmol/L) for a total of 23 episodes out of 4140 measurements (0.56%). Estimated creatinine clearance for those with hypoglycemia was 69 +/- 32 mL/min compared with 117 +/- 58 mL/min for the others (P < 0.01). CONCLUSION: Our protocol was safe and effective for the management of hyperglycemia in critically ill trauma patients receiving specialized nutritional support but should be used with caution in patients with renal insufficiency. PMID- 18403183 TI - The hydroxylapatite-bone interface: 10 years after implant installation. AB - Reconstruction of a severely atrophied maxilla by sinus augmentation with a mixture of hydroxylapatite (HA) granules and autologous cancellous bone is claimed to be a predictable means to facilitate implant placement. To the authors' knowledge, this is the first human histological case report of plasma sprayed HA-coated implants being loaded after sinus augmentation was performed with a mixture of HA granules and autologous bone. A patient gave consent for post-mortem analysis of his upper jaw, Ten years after successful reconstruction of his maxilla and functional implant loading, the region of augmentation showed stable contours both radiographically and histologically. In 10 years, the granules had only minimally degraded. HA granules outlined the grafted area to protect it against resorption. The plasma-sprayed HA coating on the dental implant was intact, showing 48% contact with the surrounding bone. PMID- 18403184 TI - Proton magnetic resonance spectroscopy in youth with severe mood dysregulation. AB - Increasing numbers of youth are presenting for psychiatric evaluation with markedly irritable mood plus "hyperarousal" symptoms. Diagnostically homeless in current nosology, the syndrome (as well as its underlying neurobiology) is little understood. To address this problem, we conducted an exploratory proton magnetic resonance spectroscopy (MRS) study in a large sample of youth with chronic, functionally disabling irritability accompanied by hyperarousal, a clinical syndrome known as "severe mood dysregulation" (SMD), which may represent a broad phenotype of pediatric bipolar disorder. Medication-free SMD youth (N=36) and controls (N=48) underwent 1.5 Tesla MRS in four regions of interest. The following three neurometabolites, relative to creatine (Cr), were quantified with LCModel Software: (a) myo-inositol (mI), a marker of intra-cellular second messengers linked to the neurobiology of bipolar disorder; (b) glutamate/glutamine (GLX), a marker of the major excitatory neurotransmitter glutamate; and (c) N-acetyl aspartate (NAA), a marker of neuronal energetics. SMD subjects had significantly lower temporal mI/Cr versus controls. However, this difference did not survive correction for multiple comparisons. Given studies implicating mI in lithium's action in BD adults and youth, further work is necessary to determine potential therapeutic implications of our present finding and how SMD youth differ pathophysiologically from those with strictly defined BD. PMID- 18403185 TI - Synthesis, in vitro and in vivo characterization of novel ethyl dioxy phosphate prodrug of propofol. AB - A novel ethyl dioxy phosphate prodrug of propofol (3) was synthesized and characterized in vitro and in vivo as safer alternative for phosphonooxymethyl prodrugs. The synthesis of 3 was achieved via vinyl and 1-chloroethyl ether intermediates, followed by addition of phosphate group. Aqueous solubility and chemical stability of 3 was determined in buffer solutions and the bioconversion of 3 to propofol was determined in vitro and in vivo. The results show that 3 greatly enhanced the aqueous solubility of propofol (solubility over 10 mg/mL) and the stability in buffer solution (t1/2=5.2+/-0.2 days at pH 7.4, r.t.) was sufficient for i.v. administration. The enzymatic hydrolysis of 3 to propofol was extremely rapid in vitro (t1/2=21+/-3s) and 3 was readily converted to propofol in vivo in rats. During bioconversion, 3 releases acetaldehyde, a less toxic compound than the formaldehyde released from the phosphonooxymethyl prodrug of propofol (Aquavan), currently undergoing clinical trials. The maximum plasma concentration of propofol, 3.0+/-0.2 microg/mL, was reached within 2.1+/-0.8 min after the i.v. administration of 3. The present study indicates that ethyl dioxy phosphate represents a potentially useful water-soluble prodrug structure suitable for i.v. administration. PMID- 18403186 TI - [Obstructive congenital multiple epulis. A case report in a male neonate]. AB - Congenital epulis or congenital gingival cell tumour is a rare tumour in the neonate. It arises from the gingival mucosa. This benign condition can be life threatening when obstructive. In this report, a case of a male neonate, with features of obstructive congenital epulis arising from both maxillary and mandibular alveolar ridge, interfering with breast-feeding is described. Diagnosis was suspected clinically and confirmed by histology. Outcome was good after prompt surgery. PMID- 18403187 TI - A new genus, Helgoeca gen. nov., for a nudiform choanoflagellate. AB - A new genus, Helgoeca gen. nov., has been designated to accommodate a nudiform loricate choanoflagellate (American Type Culture Collection strain ATCC 50073) that was incorrectly attributed to the tectiform genus Acanthoecopsis (=Acanthocorbis). The first indication that this species might be nudiform came from a four-gene phylogeny of the choanoflagellates which recovered ATCC 50073 within a strongly supported monophyletic clade comprising two other nudiform taxa. Fortunately an isolate of the species in question was available from the ATCC and when observed in rapidly growing culture it was immediately apparent that this species divided with the production of 'naked' motile cells; a typically nudiform character. The beaker-shaped lorica of this species consists of an outer layer of approximately 11 longitudinal costae, which terminate anteriorly as spines, and an equal or larger number of helical costae, with a left-handed conformation, each of which terminates anteriorly adjacent to the base of a spine. The pattern of costae in this species is indistinguishable from that of Acanthocorbis nana Thomsen and for this reason A. nana has been transferred to the new genus Helgoeca gen. nov., as the type species. PMID- 18403188 TI - Ultrastructure and molecular phylogeny of Stephanopogon minuta: an enigmatic microeukaryote from marine interstitial environments. AB - Although Stephanopogon was described as a putative ciliate more than a century ago, its phylogenetic position within eukaryotes has remained unclear because of an unusual combination of morphological characteristics (e.g. a highly multiflagellated cell with discoidal mitochondrial cristae). Attempts to classify Stephanopogon have included placement with the Ciliophora, the Euglenozoa, the Heterolobosea and the Rhizaria. Most systematists have chosen, instead, to conservatively classify Stephanopogon as incertae sedis within eukaryotes. Despite the obvious utility of molecular phylogenetic data in resolving this issue, DNA sequences from Stephanopogon have yet to be published. Accordingly, we characterized the molecular phylogeny and ultrastructure of Stephanopogon minuta, a species we isolated from marine sediments in southern British Columbia, Canada. Our results showed that S. minuta shares several features with heteroloboseans, such as discoidal mitochondrial cristae, a heterolobosean-specific (17_1 helix) insertion in the small subunit ribosomal RNA gene (SSU rDNA) and the lack of canonical Golgi bodies. Molecular phylogenetic analyses of SSU rDNA demonstrated that S. minuta branches strongly within the Heterolobosea and specifically between two different tetraflagellated lineages, both named 'Percolomonas cosmopolitus.' Several ultrastructural features shared by S. minuta and P. cosmopolitus reinforced the molecular phylogenetic data and confirmed that Stephanopogon is a highly divergent multiflagellated heterolobosean that represents an outstanding example of convergent evolution with benthic eukaryovorous ciliates (Alveolata). PMID- 18403189 TI - Influence of very long-chain n-3 fatty acids on plasma markers of inflammation in middle-aged men. AB - This study investigated the effects of a moderate dose of long-chain n-3 polyunsaturated fatty acids (1.8 g eicosapentaenoic acid (EPA) plus 0.3g docosahexaenoic acid (DHA) per day) given for 8 weeks to healthy middle-aged males on cardiovascular risk factors, particularly plasma lipids and inflammatory markers. The study was double-blind and placebo-controlled. The proportion of EPA was significantly increased in plasma phosphatidylcholine (from 1.4% to 5.0% of total fatty acids; P<0.001), cholesteryl esters (from 1.2% to 4.5%; P<0.001) and triacylglycerols (from 0.3% to 1.8%; P<0.001). In contrast, the more modest increases in DHA in these lipid fractions were not significant. There was very little effect of n-3 fatty acids on the risk factors measured, apart from a reduction in plasma soluble intercellular adhesion molecule (sICAM)-1 concentration compared with placebo (P=0.05). The change in plasma sICAM-1 concentration was significantly inversely related to the change in DHA in plasma phosphatidylcholine (r=-0.675; P=0.001), but less so to the change in EPA (r= 0.406; P=0.076). Data from the present study suggest that marine oil providing 1.8 g of EPA plus 0.3g DHA/day is not sufficient to demonstrate marked effects on cardiovascular risk factors (plasma lipids and inflammatory markers) in healthy middle-aged men, although there may be a slight anti-inflammatory effect as indicated by the decrease in sICAM-1. The stronger association between changes in DHA than EPA and sICAM-1 concentrations suggest that DHA may be more anti inflammatory than EPA. Thus, one reason why only limited effects were seen here may be that the dose of DHA provided was insufficient. PMID- 18403190 TI - Thymoproteasome: probable role in generating positively selecting peptides. AB - The proteasome is the protein destroying machinery conserved in all eukaryotes and plays essential roles in various cellular processes. Apart from the conserved 'standard' proteasome, a special type of proteasome called 'immunoproteasome' exists in vertebrates for better presentation of antigenic peptides on MHC class I molecules. Recently, another vertebrate-specific proteasome was discovered in the thymus. This 'thymoproteasome' has a novel catalytic subunit 'beta5t' with unusual enzymatic activity and is expressed exclusively in cortical thymic epithelial cells (cTECs), which catalyze positive selection of developing thymocytes. beta5t-deficient mice exhibit severe impairment in CD8(+) T cell development. These findings suggest that cTECs are quite unique cells capable of presenting a unique set of self-peptides that are not seen in other cells and are required for positive selection of CD8(+) T cells. PMID- 18403191 TI - Thymus development and function. AB - Thymopoiesis is a complex and highly dynamic process. It involves reciprocal tissue interactions between epithelial cells derived from the endoderm of the anterior foregut and neural crest-derived mesenchyme to form the thymic rudiment. This organ anlage attracts thymocyte progenitors and supports their differentiation and functional maturation into a self-tolerant diverse repertoire of T cells. In recent years, a more detailed picture of the molecular mechanisms determining the formation of the thymic rudiment and those controlling the maturation of the epithelial compartment has emerged and these are briefly summarized here. This review also addresses new experimental approaches toward a better understanding of thymopoiesis and discusses the impact of new animal models. PMID- 18403192 TI - Genetically determined lymphopenia and autoimmune manifestations. AB - Hypomorphic defects of V(D)J recombination in humans lead to residual T cell development. In these lymphopenic conditions, homeostatic lymphocyte proliferation occurs, and key mechanisms that normally maintain host tolerance are altered, allowing peripheral expansion of oligoclonal and autoreactive T cells. Recently described murine models support this notion. This review describes human and murine situations, in which genetically determined T and B cell lymphopenia is associated with autoimmune manifestations. PMID- 18403193 TI - Detecting AhR ligands in sediments using bioluminescent reporter yeast. AB - Sediments polluted with high concentrations of persistent organic pollutants, many of which are ligands of the aryl hydrocarbon receptor (AhR), are currently of concern around the industrialized world. Bioassays that can detect the presence of AhR ligands in environmental samples offer a relatively rapid and cost-effective means of prioritizing samples before more elaborate, laborious, and costly chemical analyses are applied. This paper presents a new bioluminescent yeast assay based on transcriptional activation of AhR. Its applicability for determining AhR ligands in complex environmental samples was demonstrated by analyzing a set of sediment samples from the River Kymi, Finland. The results from the assay are shown to be consistent with those from both a chemical analysis and an H4IIE-luc bioassay. The yeast assay procedure is simple and can be performed within 1 day. The yeasts grow rapidly, are easy to handle, and do not require continuous cell culturing. Moreover, the robustness of the yeast allows the application of the test to crude extracts or even sediment suspensions. The yeast assay described in this paper can be useful in screening and prioritization of samples prior to chemical analysis. Moreover, the strain can be used in the construction of fibre-optic biosensors. PMID- 18403194 TI - Synthesis of a new long-wavelength latent fluorimetric indicator for analytes determination in the DT-Diaphorase coupling dehydrogenase assay system. AB - We synthesized a new long-wavelength latent fluorogenic probe BQC (1) to monitor DTD activity. The fluorogenic chemical transformation of BQC triggered by DTD in the presence of NADH is through a series of tandem reactions, DTD-catalyzed benzoquinone reduction, trimethyl-locks cyclization and intramolecular urea formation, which are spontaneous and irreversible at physiological temperature in aqueous media. The fluorescence signal revealed by this process is specific and exhibited in the near red spectrum region with emission maxima at 595 nm, and it could be competitively inhibited by menadione. The fluorescent response of BQC is insensitive to various biological thiol reductants. Furthermore, pro-fluorophore BQC is a sensitive fluorimetric indicator for analytes determination in the oxygen-insensitive DTD-coupled dehydrogenases assay by including NAD(+) which will convert to NADH by reaction in the presence of analytes. This novel oxygen insensitive assay demonstrates a good relationship in detecting 3-hydroxybutyrate and glucose-1-phosphate in 1-10 microM range, which presents to the applicability for the construction of fiber-optic biosensors in the future clinical diagnostic. PMID- 18403195 TI - A possibility of detection of the non-charge based analytes using ultra-thin body field-effect transistors. AB - Ultra-thin body of p-type field-effect transistors were developed as transducer for biosensors. Changes of conductance resulted from the changes of the surface potentials of ultra-thin body field-effect transistors (UTB-FETs) due to surface chemical modifications were demonstrated. The channel surface of UTB-FETs were modified with N-[3-(trimethoxysilyl)propyl]ethylenediamine (AEAPTMS) and then gold nanoparticles (AuNPs) to immobilize the bio-component, the genetically engineered Delta(5)-3-ketosteroid isomerase (Art_KSI) or the Art_KSI conjugated with charged reporter (Art_KSI_mA51). The binding of charge-based molecules or nanoparticles has been demonstrated to strongly affect the conductivity of UTB FETs; the increase or decrease of the conductance depends on the polarity of the immobilized molecules or nanoparticles. A new protocol involving the detection of a non-charged analyte relied on the competitive binding of analyte (19 norandrostendione) and a charged reporter (mA51) with KSI. When exposed to a 19 norandrostendione solution (10 microM), the conductance of Art_KSI_mA51-modified UTB-FET increased by 265 nS ( approximately 12%). On the other hand, conductance of Art_KSI-modified UTB-FET showed no distinct change under the same detection conditions. PMID- 18403196 TI - Emerging strategies of lignin engineering and degradation for cellulosic biofuel production. AB - Ethanol and other biofuels produced from lignocellulosic biomass represent a renewable, more carbon-balanced alternative to both fossil fuels and corn-derived or sugarcane-derived ethanol. Unfortunately, the presence of lignin in plant cell walls impedes the breakdown of cell wall polysaccharides to simple sugars and the subsequent conversion of these sugars to usable fuel. Recent advances in the understanding of lignin composition, polymerization, and regulation have revealed new opportunities for the rational manipulation of lignin in future bioenergy crops, augmenting the previous successful approach of manipulating lignin monomer biosynthesis. Furthermore, recent studies on lignin degradation in nature may provide novel resources for the delignification of dedicated bioenergy crops and other sources of lignocellulosic biomass. PMID- 18403198 TI - Sexual differentiation of the rodent hypothalamus: hormonal and environmental influences. AB - Brain sexual differentiation is a complex developmental phenomenon influenced by the genetic background, sex hormone secretions and environmental inputs, including pollution. The main hormonal drive to masculinize and defeminize the rodent brain is testosterone secreted by the testis. The hormone does not influence sex brain differentiation only in its native configuration, but it mostly needs local conversion into active metabolites (estradiol and DHT) through the action of specific enzymatic systems: the aromatase and 5alpha-reductase (5alpha-R), respectively. This allows the hormone to control target cell gene expression either through the estrogen (ER) or the androgen (AR) receptors. The developmental profile of testosterone metabolizing enzymes, different in the two sexes, is therefore of the utmost importance in affecting the bioavailability of the steroids active in brain differentiation. Widely diffused pollutants, like polychlorinated biphenyls (PCBs) are able to affect the production and/or action of testosterone metabolites, exerting detrimental influences on reproduction and sex behavior. The main studies performed in our and other laboratories concerning the pattern of expression and the control of the enzymatic systems involved in brain androgen action and metabolism are shortly reviewed. Some recent data on the influence exerted by PCBs on these metabolic systems are also reported. PMID- 18403197 TI - Towards atomic resolution structural determination by single-particle cryo electron microscopy. AB - Recent advances in cryo-electron microscopy and single-particle reconstruction (collectively referred to as 'cryoEM') have made it possible to determine the three-dimensional (3D) structures of several macromolecular complexes at near atomic resolution ( approximately 3.8-4.5A). These achievements were accomplished by overcoming the challenges in sample handling, instrumentation, image processing, and model building. At near-atomic resolution, many detailed structural features can be resolved, such as the turns and deep grooves of helices, strand separation in beta sheets, and densities for loops and bulky amino acid side chains. Such structural data of the cytoplasmic polyhedrosis virus (CPV), the Epsilon 15 bacteriophage and the GroEL complex have provided valuable constraints for atomic model building using integrative tools, thus significantly enhancing the value of the cryoEM structures. The CPV structure revealed a drastic conformational change from a helix to a beta hairpin associated with RNA packaging and replication, coupling of RNA processing and release, and the long sought-after polyhedrin-binding domain. These latest advances in single-particle cryoEM provide exciting opportunities for the 3D structural determination of viruses and macromolecular complexes that are either too large or too heterogeneous to be investigated by conventional X-ray crystallography or nuclear magnetic resonance (NMR) methods. PMID- 18403199 TI - What are comparative studies telling us about the mechanism of ERbeta action in the ERE-dependent E2 signaling pathway? AB - Estrogen hormone (E2) signaling is primarily conveyed by the estrogen receptors (ER) alpha and beta. ERs are encoded by two distinct genes and share varying degrees of domain-specific structural/functional similarities. ERs mediate a complex array of nuclear and non-nuclear events critical for the homeodynamic regulation of various tissue functions. The canonical nuclear signaling involves the interaction of ERalpha and ERbeta with specific DNA sequences, the so-called estrogen responsive elements (EREs). This interaction constitutes the initial step in ERE-dependent signaling in which ERbeta is a weaker transcription factor than ERalpha in response to E2. However, it remains unclear why transactivation potencies of ER subtypes differ. Studies suggest that the amino-terminus, the least conserved structural region, of ERbeta, but not that of ERalpha, impairs the ability of the receptor to bind to ERE independent of E2. Although the impaired ERbeta-ERE interaction contributes, it is not sufficient to explain the weak transactivation potency of the receptor. It appears that the lack of transactivation ability and of the capability of the amino-terminus of ERbeta, as opposed to that of ERalpha, to functionally interact with the carboxyl-terminal hormone-dependent activation domain is also critical for the receptor-specific activity. Thus, the structurally distinct amino-termini of ERs are important determinants in defining the function of ER-subtypes in the ERE-dependent pathway. This could differentially affect the physiology and pathophysiology of E2 signaling. PMID- 18403200 TI - Cationized starch-based material as a new ion-exchanger adsorbent for the removal of C.I. Acid Blue 25 from aqueous solutions. AB - This article describes the use of a cationized starch-based material as new ion exchanger adsorbent for the removal of C.I. Acid Blue 25 (AB 25) from aqueous solutions. Batch adsorption studies concerning the effects of contact time, pH and temperature are presented and discussed. Adsorption experimental data showed that: (i) the process was uniform and rapid: adsorption of dye reached equilibrium in 50 min in the wide pH range of dye solutions; (ii) adsorption kinetics followed the pseudo-second order model; (iii) the Langmuir model yielded a much better fit than the Freundlich model for the dye concentration range under study; (iv) this adsorbent exhibited interesting adsorption capacities: on the basis of the Langmuir analysis, the maximum adsorption capacity was determined to be 322 mg of dye per gram of material at 25 degrees C; (v) the adsorption capacity decreased with increasing temperature; and (vi) the negative value of free energy change indicated the spontaneous nature of adsorption. PMID- 18403201 TI - The color-vision circuit in the medulla of Drosophila. AB - BACKGROUND: Color vision requires comparison between photoreceptors that are sensitive to different wavelengths of light. In Drosophila, this is achieved by the inner photoreceptors (R7 and R8) that contain different rhodopsins. Two types of comparisons can occur in fly color vision: between the R7 (UV sensitive) and R8 (blue- or green sensitive) photoreceptor cells within one ommatidium (unit eye) or between different ommatidia that contain spectrally distinct inner photoreceptors. Photoreceptors project to the optic lobes: R1-R6, which are involved in motion detection, project to the lamina, whereas R7 and R8 reach deeper in the medulla. This paper analyzes the neural network underlying color vision into the medulla. RESULTS: We reconstruct the neural network in the medulla, focusing on neurons likely to be involved in processing color vision. We identify the full complement of neurons in the medulla, including second-order neurons that contact both R7 and R8 from a single ommatidium, or contact R7 and/or R8 from different ommatidia. We also examine third-order neurons and local neurons that likely modulate information from second-order neurons. Finally, we present highly specific tools that will allow us to functionally manipulate the network and test both activity and behavior. CONCLUSIONS: This precise characterization of the medulla circuitry will allow us to understand how color vision is processed in the optic lobe of Drosophila, providing a paradigm for more complex systems in vertebrates. PMID- 18403202 TI - Organelles in Blastocystis that blur the distinction between mitochondria and hydrogenosomes. AB - Blastocystis is a unicellular stramenopile of controversial pathogenicity in humans. Although it is a strict anaerobe, Blastocystis has mitochondrion-like organelles with cristae, a transmembrane potential and DNA. An apparent lack of several typical mitochondrial pathways has led some to suggest that these organelles might be hydrogenosomes, anaerobic organelles related to mitochondria. We generated 12,767 expressed sequence tags (ESTs) from Blastocystis and identified 115 clusters that encode putative mitochondrial and hydrogenosomal proteins. Among these is the canonical hydrogenosomal protein iron-only [FeFe] hydrogenase that we show localizes to the organelles. The organelles also have mitochondrial characteristics, including pathways for amino acid metabolism, iron sulfur cluster biogenesis, and an incomplete tricarboxylic acid cycle as well as a mitochondrial genome. Although complexes I and II of the electron transport chain (ETC) are present, we found no evidence for complexes III and IV or F1Fo ATPases. The Blastocystis organelles have metabolic properties of aerobic and anaerobic mitochondria and of hydrogenosomes. They are convergently similar to organelles recently described in the unrelated ciliate Nyctotherus ovalis. These findings blur the boundaries between mitochondria, hydrogenosomes, and mitosomes, as currently defined, underscoring the disparate selective forces that shape these organelles in eukaryotes. PMID- 18403203 TI - Male fertility of malaria parasites is determined by GCS1, a plant-type reproduction factor. AB - Malaria, which is caused by Plasmodium parasites, is transmitted by anopheline mosquitoes. When gametocytes, the precursor cells of Plasmodium gametes, are transferred to a mosquito, they fertilize and proliferate, which render the mosquito infectious to the next vertebrate host. Although the fertilization of malaria parasites has been considered as a rational target for transmission blocking vaccines, the underlying mechanism is poorly understood. Here, we show that the rodent malaria parasite gene Plasmodium berghei GENERATIVE CELL SPECIFIC 1 (PbGCS1) plays a central role in its gametic interaction. PbGCS1 knockout parasites show male sterility, resulting in unsuccessful fertilization. Because such a male-specific function of GCS1 has been observed in angiosperms, this indicates, for the first time, that parasite sexual reproduction is controlled by a machinery common to flowering plants. Our present findings provide a new viewpoint for understanding the parasitic fertilization system and important clues for novel strategies to attack life-threatening parasites. PMID- 18403204 TI - Patient's self-determination in intensive care-from an action- and confirmation theoretical perspective. The intensive care nurse view. AB - When becoming an intensive care patient life changes dramatically. In order to save life, different actions are performed by the caregivers and the patient's ability to exercise self-determination is non-existent. After the acute phase the patient is more awake and the possibilities for self-determination change. The purpose of this study was to describe intensive care nurses' (ICNs) views of patient's self-determination in an intensive care unit and to systematize ICNs' nursing actions for supporting patient's self-determination from an action- and confirmation-theoretic perspective. In order to answer these questions, 17 interviews with ICNs were conducted by the use of the Critical Incident Technique (CIT). The transcripts were then analysed using a hermeneutic analysis method and structured by the SAUC model for confirming nursing. The main findings were that the ICN thought that the ICU patient's self-determination was low and restricted. It was more common that the ICN acted to strengthen the patient's self determination in nursing care, but there were no specific nursing goals for patient's self-determination. The most common actions for supporting self determination were supplying the patient with information and engaging the patient in making a day plan. The nursing implications are that the ICN's view of human being as an acting subject is important for the ICN's awareness to recognise the patient's own personal resources to handle the critically ill situation and that the ICN's competence to manifest qualified nursing is necessary for strengthening patient's self-determination. PMID- 18403206 TI - Stabilization of a multi-segment model of bipedal standing by local joint control overestimates the required ankle stiffness. PMID- 18403205 TI - Oral care and the risk of bloodstream infections in mechanically ventilated adults: A review. AB - RATIONALE: Bacteraemia, defined as the presence of viable bacteria in the circulating blood can result in bloodstream infection, which is one of the most frequent and challenging hospital-acquired infections. Bacteraemia occurs in healthy populations with manipulation of the oral mucosa, including toothbrushing. Oral care is commonly administered to mechanically ventilated patients, it is important to determine whether this practice contributes to the incidence of bacteraemia. This paper reviews the literature on the link between the manipulation of the oral cavity and the development of bacteraemia in mechanically ventilated adults. METHODS: Searches were conducted using Medline, CINAHL, and the Cochrane Library databases. Article inclusion criteria were (1) a focus on mechanical ventilation and critical illness, (2) human subjects, (3) adult subjects, and (4) publication in English (or available English translation). RESULTS: Nine articles met inclusion criteria and were critiqued. All relied upon clinical data as outcome measures; many were retrospective. The three organisms most often associated with nosocomial bloodstream infections were Staphylococcus aureus, coagulase negative staphylococci, and Enterococcus species. Establishing the origin of bacteraemia was problematic in most studies. CONCLUSIONS: Additional research is needed to understand the relationship of oral care practices to bacteraemia in mechanically ventilated adults. PMID- 18403207 TI - Attacking the defenders: plant viruses fight back. AB - Plants use RNA silencing mechanisms and produce short-interfering RNA (siRNA) molecules in a defense response against viral infection. To counter this defense response, viruses produce suppressor proteins, which can block the host silencing pathway or interfere with its function in plant cells. The targets for many viral suppressors and the mechanisms by which they function in plant cells are still largely unknown. Recent reports describe that the 2b suppressor of the Cucumber mosaic virus binds ARGONAUTE and that the P0 suppressor of Polerovirus targets ARGONAUTE to degradation. Another report has revealed that the V2 suppressor of tomato yellow mosaic virus binds the coiled-coil protein suppressor of the gene silencing SGS3 homolog. These reports provide novel insight into the mechanisms developed by viruses to disable the defense system of the plant. PMID- 18403208 TI - Linear accelerator radiosurgery for treatment of vestibular schwannomas in neurofibromatosis 2. AB - Management of vestibular schwannomas in patients with neurofibromatosis 2 (NF2) balances growth control against preservation of hearing with the primary aim of maintaining patient quality of life. Surgical resection of these lesions carries greater risk of functional deterioration than in sporadic cases. Stereotactic radiosurgery is a less invasive option that provides comparable, if not superior outcomes to resection. Previous studies on the efficacy of stereotactic radiosurgery for vestibular schwannomas in NF2 have reported results from delivery by Gamma Knife systems. The efficacy of linear accelerator (LINAC) delivered treatment has not been specifically addressed. Modelling studies suggest that lesional conformality is superior with Gamma Knife, but clinical studies on sporadic vestibular schwannomas show equivalent results between the two systems. Our experience with LINAC radiosurgery in NF2 reported here shows good long-term growth control in four patients with vestibular schwannomas. PMID- 18403210 TI - Principles of demineralization: modern strategies for the isolation of organic frameworks. Part I. Common definitions and history. AB - In contrast to biomineralization phenomena, that are among the most widely studied topics in modern material and earth science and biomedicine, much less is systematized on modern view of demineralization. Biomineralized structures and tissues are composites, containing a biologically produced organic matrix and nano- or microscale amorphous or crystalline minerals. Demineralization is the process of removing the inorganic part, or the biominerals, that takes place in nature via either physiological or pathological pathways in organisms. In vitro demineralization processes, used to obtain mechanistic information, consist in the isolation of the mineral phase of the composite biomaterials from the organic matrix. Physiological and pathological demineralization include, for example, bone resorption mediated by osteoclasts. Bioerosion, a more general term for the process of deterioration of the composite biomaterials represents chemical deterioration of the organic and mineral phase followed by biological attack of the composite by microorganisms and enzymes. Bioerosional organisms are represented by endolithic cyanobacteria, fungi, algae, plants, sponges, phoronids and polychaetes, mollusks, fish and echinoids. In the history of demineralization studies, the driving force was based on problems of human health, mostly dental caries. In this paper we summarize and integrate a number of events, discoveries, milestone papers and books on different aspect of demineralization during the last 400 years. Overall, demineralization is a rapidly growing and challenging aspect of various scientific disciplines such as astrobiology, paleoclimatology, geomedicine, archaeology, geobiology, dentistry, histology, biotechnology, and others to mention just a few. PMID- 18403209 TI - A SIM-ultaneous role for SUMO and ubiquitin. AB - Ubiquitin and ubiquitin-like proteins (Ubls) share a beta-GRASP fold and have key roles in cellular growth and suppression of genome instability. Despite their common fold, SUMO and ubiquitin are classically portrayed as distinct, and they can have antagonistic roles. Recently, a new family of proteins, the small ubiquitin-related modifier (SUMO)-targeted ubiquitin ligases (STUbLs), which directly connect sumoylation and ubiquitylation, has been discovered. Uniquely, STUbLs use SUMO-interaction motifs (SIMs) to recognize their sumoylated targets. STUbLs are global regulators of protein sumoylation levels, and cells lacking STUbLs display genomic instability and hypersensitivity to genotoxic stress. The human STUbL, RNF4, is implicated in several diseases including cancer, highlighting the importance of characterizing the cellular functions of STUbLs. PMID- 18403211 TI - Development of a low-level background gamma-ray spectrometer by KRISS. AB - A new low-level background and high-efficiency gamma-ray spectrometric system, to be used mainly for the activity certification of natural-matrix certified reference materials (CRMs) and environmental reference materials (RMs) that has been developed on the grounds of the Korea Research Institute of Standards and Science (KRISS). The spectrometer consists of a low-background high-purity germanium detector with a relative efficiency of 120% and various shielding devices to reduce radiation background. The cabinet-shaped device made of 10ton of shielding materials encloses the germanium detector for protection against background from natural radioactivity and neutrons. Three plates of 50-mm-thick plastic scintillation detectors on top of the passive shielding cabinet suppress cosmogenic background by detecting high-energetic cosmic muons bombarding the germanium detector. The measured background rate of the spectrometer for the energy range 50-3000keV was 1.72s(-1). PMID- 18403212 TI - Sertraline slows disease progression and increases neurogenesis in N171-82Q mouse model of Huntington's disease. AB - Huntington's disease (HD) is an inherited progressive neurodegenerative disorder resulting from CAG repeat expansion in the gene that encodes for the protein huntingtin. To identify neuroprotective compound (s) that can slow down disease progression and can be administered long term with few side effects in Huntington's disease, we investigated the effect of sertraline, a selective serotonin reuptake inhibitor (SSRI) which has been shown to upregulate BDNF levels in rodent brains. We report here that in HD mice sertraline increased BDNF levels, preserved chaperone protein HSP70 and Bcl-2 levels in brains, attenuated the progression of brain atrophy and behavioral abnormalities and thereby increased survival. Sertraline also enhanced neurogenesis, which appeared to be responsible for mediating the beneficial effects of sertraline in HD mice. Additionally, the effective levels of sertraline are comparable to the safe levels achievable in humans. The findings suggest that sertraline is a potential candidate for treatment of HD patients. PMID- 18403213 TI - Dislocation of an upper third molar by an ossifying fibroma--case report. AB - BACKGROUND: The occurrence of ossifying fibromas (OFs) in childhood and adolescence has been described in the literature, along with different courses of the disease due to different growth rates. CASE REPORT: In the case of the 15 year-old female patient presented here, an OF resulted in displacement of a maxillary third molar far into the maxillary sinus. It is assumed that the tumour originated coronal to the affected tooth 18. Radiographs document an initial rapid growth of this tumour over a period of 2 years, while its growth almost completely ceased in the next 2 years immediately prior to diagnosis and surgical treatment. The operation was complicated by unexpected profuse bleeding from the tumour tissue. CONCLUSION: The peculiarity of the OF in the case presented here is its similarity, in terms of clinical and radiological appearances, with a follicular cyst, its unusual place of origin that resulted in the migration of the tooth 18 into the maxillary sinus, its different growth dynamics, and the pronounced haemorrhage encountered as the tumour was surgically removed. PMID- 18403214 TI - Automated, feature-based image alignment for high-resolution imaging mass spectrometry of large biological samples. AB - High-resolution imaging mass spectrometry of large biological samples is the goal of several research groups. In mosaic imaging, the most common method, the large sample is divided into a mosaic of small areas that are then analyzed with high resolution. Here we present an automated alignment routine that uses principal component analysis to reduce the uncorrelated noise in the imaging datasets, which previously obstructed automated image alignment. An additional signal quality metric ensures that only those regions with sufficient signal quality are considered. We demonstrate that this algorithm provides superior alignment performance than manual stitching and can be used to automatically align large imaging mass spectrometry datasets comprising many individual mosaic tiles. PMID- 18403215 TI - Loss of Fyn tyrosine kinase on the C57BL/6 genetic background causes hydrocephalus with defects in oligodendrocyte development. AB - The supportive functions of oligodendrocytes are required for the survival and development of axons, ensuring the organization of highly specialized neuronal networks in brain. Although the molecules that regulate oligodendrocyte differentiation in vitro have been identified, their roles in vivo are largely uncertain. Here we report that fyn deficiency on the C57BL/6 genetic background resulted in premature death, showing severe hydrocephalus with neonatal onset. One week after birth, fyn-deficient mice showed enlarged lateral ventricles with thinner cerebral cortices and degenerating axons in the corpus callosum. In addition, before the onset of myelination, the number of oligodendrocytes was reduced and their morphogenesis was impaired in the cerebral cortex. These results demonstrate that Fyn is essential for normal brain development and suggest that defects in oligodendrocyte development cause degeneration of cortical axons and subsequent hydrocephalus in fyn-deficient mice. PMID- 18403216 TI - EphB receptors co-distribute with a nicotinic receptor subtype and regulate nicotinic downstream signaling in neurons. AB - Activation of nicotinic acetylcholine receptors (nAChRs) on neurons engages calcium-dependent signaling pathways regulating numerous events. Receptors containing alpha7 subunits (alpha7-nAChRs) are prominent in this because of their abundance and high relative calcium permeability. We show here that EphB2 receptors are co-localized with postsynaptic alpha7-nAChRs on chick ciliary ganglion neurons and that treatment of the cells with an ephrinB1 construct to activate the EphB receptors exerts physical restraints on both classes of receptors, diminishing their dispersal after spine retraction or lipid raft disruption. Moreover, the ephrinB1/EphB receptor complex specifically enhances the ability of alpha7-nAChRs to activate the transcription factor CREB, acting through a pathway including a receptor tyrosine kinase, a Src family member, PI3 kinase, and protein kinase A most distally. The enhancement does not appear to result from a change in the alpha7-nAChR current amplitude, suggesting a downstream target. The results demonstrate a role for ephrin/EphB action in nicotinic signaling. PMID- 18403217 TI - EEG default mode network in the human brain: spectral regional field powers. AB - Eyes-closed (EC) and eyes-open (EO) are essential behaviors in mammalians, including man. At resting EC-EO state, brain activity in the default mode devoid of task-demand has recently been established in fMRI. However, the corresponding comprehensive electrophysiological conditions are little known even though EEG has been recorded in humans for nearly 80 years. In this study, we examined the spatial characteristics of spectral distribution in EEG field powers, i.e., sitting quietly with an EC and EO resting state of 3 min each, measured with high density 128-ch EEG recording and FFT signal analyses in 15 right-handed healthy college females. Region of interest was set at a threshold at 90% of the spectral effective value to delimit the dominant spatial field power of effective energy in brain activity. Low-frequency delta (0.5-3.5 Hz) EEG field power was distributed at the prefrontal area with great expansion of spatial field and enhancement of field power (t=-2.72, p<0.02) from the EC to the EO state. Theta (4-7 Hz) EEG field power was distributed over the fronto-central area and leaned forward from EC to the EO state but with drastic reduction in field power (t=4.04, p<0.01). The middle-frequency alpha-1 (7.5-9.5 Hz) and alpha-2 (10-12 Hz) EEG powers exhibited bilateral distribution over the posterior areas with an anterior field in lower alpha-1. Both showed significantly reduction of field powers (respectively, W=120, p<0.001 for alpha-1; t=4.12, p<0.001 for alpha-2) from EC to the EO state. Beta-1 (13-23 Hz) exhibited a similar spatial region over the posterior area as in alpha-2 and showed reduction of field power (t=4.42, p<0.001) from EC to the EO state. In contrast, high-frequency beta-2 and gamma band exhibited similar, mainly prefrontal distribution in field power, and exhibited no change from EC to the EO state. Corresponding correlation analyses indicated significant group association between EC and EO only in the field powers of delta (r=0.95, p<0.001) and theta (r=0.77, p<0.001) band. In addition, the great inter-individual variability (90 folds in alpha-1, 62 folds in alpha-2) in regional field power was largely observed in the EC state (10 folds) than the EO state in subjects. To summarize, our study depicts a network of spectral EEG activities simultaneously operative at well defined regional fields in the EC state, varying specifically between EC and EO states. In contrast to transient EEG spectral rhythmic dynamics, current study of long-lasting (e.g. 3 min) spectral field powers can characterize state features in EEG. The EEG default mode network (EEG-DMN) of spectral field powers at rest in the respective EC or EO state is valued to serve as the basal electrophysiological condition in human brain. In health, this EEG-DMN is deemed essential for evaluation of brain functions without task demands for gender difference, developmental change in age span, and brain response to task activation. It is expected to define brain dysfunction in disease at resting state and with consequences for sensory, affective and cognitive alteration in the human brain. PMID- 18403218 TI - Switching from pathogenetic treatment with alpha-lipoic acid to gabapentin and other analgesics in painful diabetic neuropathy: a real-world study in outpatients. AB - In this retrospective real-world study, we aimed to evaluate whether switching from the pathogenetic treatment option alpha-lipoic acid to drugs for symptomatic treatment of neuropathic pain such as gabapentin would be associated with changes in efficacy, safety, and cost-effectiveness. A cohort of 443 diabetic patients with chronic painful neuropathy were treated with alpha-lipoic acid 600 mg qd orally for a mean period of 5 years. After stopping this treatment, 293 patients were switched to gabapentin (600-2400 mg/day), while 150 patients remained untreated because of no acute symptoms. In the untreated group, 110 (73%) patients developed neuropathic symptoms as soon as 2 weeks after the end of treatment with alpha-lipoic acid. In the group started on gabapentin, 131 (45%) patients had to stop taking the drug due to intolerable side effects. Among the patients treated with gabapentin 132 (45%) were responders on an average dose of 1200 mg/day, whereas 161 (55%) were nonresponders at gabapentin doses up to 2400 mg/day. These patients required an alternative treatment which consisted of pregabalin, carbamazepine, amitriptyline, tramadol, or morphine as monotherapy or in combination. The daily costs for alpha-lipoic acid were considerably lower than those for gabapentin or several frequently used drug combinations. The frequency of outpatient visits was 3.8 times per 3 months during the treatment period with alpha-lipoic acid, while it increased to 7.9 per 3 months after switching to gabapentin or the other pain medications. In conclusion, switching from long-term treatment with alpha-lipoic acid to central analgesic drugs such as gabapentin in painful diabetic neuropathy was associated with considerably higher rates of side effects, frequencies of outpatient visits, and daily costs of treatment. The pathogenic treatment option represents for the practicing diabetologist an effective, safe, and cost-effective treatment option for the majority of patients with diabetic polyneuropathy. PMID- 18403220 TI - Predictive factors for postoperative outcome in temporal lobe epilepsy according to two different classifications. AB - PURPOSE: The determination of prognostic factors is important for predicting outcome after epilepsy surgery. We investigated the factors related to surgical outcome within a homogeneous group of patients suffering from pathologically proven mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS), and compared Engel's outcome classification system with the latest one proposed by the ILAE. METHOD: We included 109 patients with MTLE-HS who were followed-up for at least 1 year after epilepsy surgery. A retrospective chart review was performed to extract patients' demographic details, and potential pre postoperative risk factors. Outcome of surgery was defined by the Engel's and ILAE classifications. In addition, the course of prognosis was determined according to the changes in ILAE classifications on an annual basis. Univariate and multivariate logistic regression analyses were used for the latest available outcomes and two different courses of prognosis. RESULTS: The average duration of follow up was 4.78+/-2.55 years in the 109 patients with MTLE-HS. The univariate and multiple logistic regression analyses showed that the occurrence of seizures during the first month after surgery was a significant risk factor for a poor outcome. A history of trauma was also significant for patients with late recurrence of postsurgical seizures after at least 1-year seizure remission. CONCLUSION: The occurrence of seizures during the first month after surgery is a significant prognostic factor in patients with MTLE-HS. Ignoring early postoperative seizures in classification systems may result in difficulty in identifying the course of epilepsy after surgery. PMID- 18403219 TI - Frequency-modulated electromagnetic neural stimulation enhances cutaneous microvascular flow in patients with diabetic neuropathy. AB - AIM: The aim of this study was to investigate the effects of frequency modulated electromagnetic neural stimulation (FREMS), a recently developed safe and effective treatment of painful diabetic neuropathy, on cutaneous microvascular function. METHODS: Thirty-one patients with painful neuropathy were enrolled in a randomised, double-blind, crossover FREMS vs. placebo study; each received two series of 10 treatments of either FREMS or placebo in random sequence within no more than 3 weeks. Patients were studied at baseline, end of FREMS and placebo series, and after 4 months of follow-up. Cutaneous blood flow was measured by laser doppler flowmetry and partial tissue tension of oxygen (TcPO2) and carbonic anhydride (TcPCO2) by oxymetry at the lower extremities in basal resting conditions and as incremental response after thermal stimulation. RESULTS: Crossover analysis showed no consistent differences between FREMS and placebo. After 4-month follow-up, a 52% increase of cutaneous blood flow was observed in resting conditions (P=.0086 vs. baseline), while no differences were observed as incremental flow after warming; compared with baseline, no significant differences were observed for TcPO2 and TcPCO2, both in resting conditions and as incremental response to warm. CONCLUSION: These results indicate that 10 treatments with FREMS may induce an enhancement of microvascular blood flow measurable at 4 months of follow-up. The findings of this study will need to be confirmed in a larger, adequately powered study (ClinicalTrial.gov Id: NCT00337324). PMID- 18403221 TI - The type II collagen fragments Helix-II and CTX-II reveal different enzymatic pathways of human cartilage collagen degradation. AB - OBJECTIVE: Cartilage degradation in osteoarthritis (OA) generates the type II collagen fragments, Helix-II and CTX-II that can be used as clinical biological markers. Helix-II and C-telopeptide of type II collagen (CTX-II) levels are associated independently with progression of OA suggesting that they may be generated through different collagenolytic pathways. In this study we analyzed the release of Helix-II and CTX-II from human cartilage collagen by the proteinases reported to play a role in cartilage degradation. METHODS: In vitro, human articular cartilage extract was incubated with activated human recombinant cathepsins (Cats) and matrix-metalloproteases (MMPs). Next, we analyzed the spontaneous release of Helix-II and CTX-II from cartilage sections of patients with knee OA who were immediately deep frozen after joint replacement to preserve endogenous enzyme activity until assay. Cartilage sections were then incubated for up to 84 h in the presence or absence of E-64 and GM6001, inhibitors of cysteine proteases and MMPs, respectively. RESULTS: In vitro, Cats K, L and S generated large amount of Helix-II, but not CTX-II. Cat B generated CTX-II fragment, but destroyed Helix-II immunoreactivity. Cat D was unable to digest intact cartilage. MMPs-1, -3, -7, -9, and -13 efficiently released CTX-II, but only small amount of Helix-II. Neither CTX-II nor Helix-II alone was able to reflect accurately the collagenolytic activity of Cats and MMPs as reflected by the release of hydroxyproline. In OA cartilage explants, E-64 blunted the release of Helix-II whereas the release of CTX-II could be completely abrogated by GM6001 and only partly by E-64. CONCLUSION: These in vitro and ex vivo experiments of human cartilage suggest that Helix-II and CTX-II could be released in part by different enzymatic pathways. Helix-II and CTX-II alone reflect only partially overall cartilage collagen degradation. These findings may explain why these two biological markers could provide complementary information on disease progression in OA. PMID- 18403222 TI - Phylogenetics and the correlates of mammalian sleep: a reappraisal. AB - The correlates of mammalian sleep have been investigated previously in at least eight comparative studies in an effort to illuminate the functions of sleep. However, all of these univariate analyses treated each species, or taxonomic Family, as a statistically independent unit, which is invalid due to the phylogenetic relationships among species. Here, we reassess these influential correlates of mammalian sleep using the formal phylogenetic framework of independent contrasts. After controlling for phylogeny using this procedure, the interpretation of many of the correlates changed. For instance, and contrary to previous studies, we found interspecific support for a neurophysiological role for rapid-eye-movement sleep, such as memory consolidation. Also in contrast to previous studies, we did not find comparative support for an energy conservation function for slow-wave sleep. Thus, the incorporation of a phylogenetic control into comparative analyses of sleep yields meaningful differences that affect our understanding of why we sleep. PMID- 18403223 TI - Bronchodilator efficacy of the fixed combination of ipratropium and albuterol compared to albuterol alone in moderate-to-severe persistent asthma. AB - BACKGROUND: The potential of anticholinergics to provide bronchodilatory benefits over short-acting beta(2)-agonists (SABA) alone in patients with moderate-to severe persistent asthma has not been well defined. METHODS: An outpatient, randomized, double-blind, single-dose, crossover study in adult asthmatics with moderate-to-severe obstruction despite treatment with inhaled corticosteroids (ICS) was conducted comparing the fixed combination of ipratropium and albuterol (IB+ALB) to albuterol alone (ALB). Serial spirometry was performed over 6h. SABA were withheld for 8h, ICS and LABA for 24h. RESULTS: A total of 113 patients were randomized, 106 completed the study (males n=47; mean+/-SD age=51+/-13 years). Mean+/-SD baseline FEV(1)=1.4+/-0.5 L (49+/-12% predicted). IB+ALB resulted in significantly greater improvements over ALB in the average improvement over baseline in FEV(1) as approximated from the area under the curve from 0 to 6h after drug administration (72 ml, p<0.01) and mean peak FEV(1) response (55 ml, p<0.01) as well as higher FEV(1) responses at individual time points from 0.5 to 6h postdose (p<0.01 for all). Time to onset of response was similar between groups but time to peak and duration of response were longer with IB+ALB versus ALB (120 versus 60 min and 245 versus 106 min, respectively). CONCLUSION: IB+ALB resulted in significantly greater improvement in FEV(1) and longer duration of response compared to ALB alone in patients with moderate-to-severe persistent asthma (Trial number: 1012.50; ClinicalTrial.gov NCT00096616). PMID- 18403225 TI - Characterization of prostanoid receptors present on adrenergic neurons innervating the porcine uterine longitudinal muscle. AB - The cyclooxygenase-prostanoid pathway regulates myometrial contractility through activation of prostanoid receptors on uterine smooth muscles. However, the possible expression of prostanoid receptors on autonomic nerves cannot be excluded completely. The aim of the present study was to clarify the presence of neural prostanoid receptors on adrenergic nerves in the porcine uterine longitudinal muscle. In [(3)H]-noradrenaline-loaded longitudinal muscle strips of porcine uterus, electrical field stimulation (EFS) evoked [(3)H]-noradrenaline release in a stimulation frequency-dependent manner. The EFS-evoked release was completely abolished in Ca(2+)-free (EGTA, 1mM) incubation medium and by tetrodotoxin or omega-conotoxin GVIA, suggesting that [(3)H]-noradrenaline was released from neural components. The EFS-evoked [(3)H]-noradrenaline release was significantly enhanced by treatment with indomethacin. In the presence of indomethacin, PGE(2) and PGF(2alpha), but not PGD(2), inhibited the EFS-evoked [(3)H]-noradrenaline release. Of synthetic prostanoid receptor agonists examined, both U46619 (TP) and sulprostone (EP(1)/EP(3)) decreased the EFS-evoked [(3)H] noradrenaline release in a concentration-dependent manner, while fluprostenol (FP), BW245C (DP) and butaprost (EP(2)) were almost ineffective. SQ29548 (TP receptor antagonist) blocked the effect of U46619, but SC19220 (EP(1) receptor antagonist) did not change the inhibition by sulprostone or PGE(2). Double immunofluorescence staining using protein gene product 9.5, tyrosine hydroxylase, EP(3) receptor and TP receptor antibodies suggested the localization of EP(3) or TP receptors on adrenergic nerves in the porcine uterus. These results indicated that neural EP(3) and TP receptors are present on adrenergic nerves of the porcine uterine longitudinal muscle. Endogenous prostanoid produced by cyclooxygenase can regulate noradrenaline release in an inhibitory manner through activation of these neural prostanoid receptors. PMID- 18403224 TI - Unraveling the complexities of sphingosine-1-phosphate function: the mast cell model. AB - Sphingosine-1-phosphate (S1P) is a lipid mediator involved in diverse biological processes, from vascular and neural development to the regulation of lymphocyte trafficking. Many of its functions are regulated by five widely expressed S1P G protein-coupled receptors (S1P(1-5)). S1P is produced mostly intracellularly, thus, much of its potential as an autocrine and paracrine mediator depends on how, when, and where it is generated or secreted out of the cells. However, S1P can also have intracellular activity independent of its receptors, adding to the complexity of S1P function. The mast cell, a major effector cell during an allergic response, has proven instrumental towards understanding the complex regulation and function of S1P. Antigen (Ag) engagement of the IgE receptor in mast cells stimulates sphingosine kinases, which generate S1P and are involved in the activation of calcium fluxes critical for mast cell responses. In addition, mast cells secrete considerable amounts of S1P upon activation, thus affecting the surrounding tissues and recruiting inflammatory cells. Export of S1P is also involved in the autocrine transactivation of S1P receptors present in mast cells. The in vivo response of mast cells, however, is not strictly dependent on their ability to generate S1P, but they are also affected by changes in S1P in the environment previous to Ag challenge. This review will discuss the recent advances towards understanding the intricacies of S1P generation, secretion and regulation in mast cells. In addition, how S1P receptors are activated and their involvement in mast cell functions will also be covered, including new insights on the role of S1P in the mast cell-mediated allergic response of systemic anaphylaxis. PMID- 18403226 TI - Causes for low positive predictive values of CD4 counts for antiretroviral treatment failure. PMID- 18403227 TI - [The rare blood groups: a public health challenge]. AB - A rare blood group is usually defined as the absence of a high prevalence antigen or the absence of several antigens within a single blood group system, if its prevalence in France is 4/1000 or less in the general population. An individual with a rare blood phenotype can develop a naturally-occurring or immune antibody corresponding to his rare specificity. In case an extremely low stock of compatible blood is available at the national level, a so-called "transfusion deadlock" is described. Most of the individuals with a rare blood group are coincidently identified when a routine pretransfusion testing or pregnancy follow up is performed, if the antibody(ies) corresponding to the rare specificity is(are) present. Other individuals are discovered following a systematic red cell typing, or family investigations in siblings. One hundred and twenty-one rare blood specificities and 42 rare blood genotypes are currently defined at the French National Reference Laboratory for Blood Groups (CNRGS-Paris). The French national registry of individuals with a rare blood phenotype/genotype includes about 9600 people, who are urged to regularly donate blood for the National Rare Blood Bank. This bank, based on a homologous blood transfusion program, is in charge of the long-term storage of rare frozen blood units, that can only be delivered after receiving authorization from the CNRGS. The global and individual care management of the individuals with a rare blood group, concerning potentially several hundred thousand people in France, requires a close cooperation between all the protagonists within the transfusion chain. PMID- 18403228 TI - Construction and infection of a new simian/human immunodeficiency chimeric virus (SHIV) containing the integrase gene of the human immunodeficiency virus type 1 genome and analysis of its adaptation to monkey cells. AB - Expanding the HIV-1-derived regions in the SHIV genome may help to clarify the viral restriction factors determining the host range. In this study, we constructed a new SHIV having the reverse transcriptase and integrase-encoding regions of HIV-1 in addition to the 3' half genomic region of HIV-1. This SHIV, termed SHIVrti/3rn, could replicate in a monkey CD4+ T cell line, HSC-F, although its replication in monkey PBMCs was very weak. After SHIVrti/3rn was passaged in HSC-F cells for 26weeks, it gradually began to replicate in monkey PBMCs. This monkey-cell-adapted virus, termed SHIVrti/3rnP, could replicate in rhesus macaques. The whole genome of SHIVrti/3rnP was sequenced and was found to differ from SHIVrti/3rn at eleven positions. We constructed a series of mutants having some or all of these mutations and investigated their replication kinetics. The mutational analysis revealed that all of the mutations, but mainly the mutations in env, were responsible for the adaptation in HSC-F cells and were enough to replicate in rhesus PBMCs. Of all the SHIVs reported so far that can infect rhesus monkeys in vivo, SHIVrti/3rnP is the one that is genetically the closest to HIV-1. PMID- 18403229 TI - CCR5 is involved in controlling the early stage of Cryptosporidium parvum infection in neonates but is dispensable for parasite elimination. AB - Chemokines play a critical role in immune cell trafficking and the transition from an innate to an acquired immune response. We analyzed host response in neonatal mice deficient in chemokine receptor CCR5 following infection with the intracellular protozoan parasite Cryptosporidium parvum. CCR5 neonatal mice had a higher parasite burden at the early stage of infection but eliminated the parasite as efficiently as their wild-type counterparts. The higher sensitivity of neonates at the beginning of infection was not due to an altered IFNgamma response. An increased CCR2-attracting chemokine response associated with the recruitment of CCR2-positive cells in the infected mucosa may have compensated for the absence of CCR5. A lack of CCR5 thus has an impact in the early stage of C. parvum infection in neonates, but this receptor is dispensable for subsequent parasite elimination. PMID- 18403230 TI - Superinfection of defective human immunodeficiency virus type 1 with different subtypes of wild-type virus efficiently produces infectious variants with the initial viral phenotypes by complementation followed by recombination. AB - Superinfection rates of human immunodeficiency virus type 1 (HIV-1) have increasingly been leading to more variation in HIV-1, as evidenced by the emergence of circulating recombinant forms (CRFs). We recently reported complementation in a persistently replication-defective subtype B-infected cell clone, L-2, by superinfection with CRF15_01B. The L-2 cells continuously produce immature particles due to a one-base insertion at pol protease. Proviruses in the superinfected cells carried both subtypes and produced particles with a mature morphology. In this study, we examined possible recombination following complementation to generate replication-competent variants by using three cell clones prepared from superinfected L-2 cells. The individual clones predominantly expressed the initial subtype B-derived mature Gag proteins. However, the viral particles carried both subtype B with the mutation and wild-type CRF15_01B at pol, suggesting the generation of virions with heterozygous RNAs. Interestingly, with cell-free passages of the progeny, defective particles disappeared, and were replaced with heterogeneous recombinants in the pol region with sequences derived from CRF15_01B that expressed subtype B phenotype. Thus, even a defective form of persistent HIV-1 can become replication-competent through superinfection-mediated complementation followed by recombination. These findings suggest the significance of long-lived infected cells as recipients for superinfection. PMID- 18403231 TI - Pbp, a cell-surface exposed plasminogen binding protein of Bacteroides fragilis. AB - The Gram-negative anaerobic bacterium B. fragilis is a member of the commensal flora of the human intestine, but is also frequently found in severe intra abdominal infections. Several B. fragilis virulence factors have been implicated in the development of these infections. A B. fragilis protein of circa 60-kDa was identified as a putative plasminogen binding protein (Pbp). The corresponding gene was located, cloned, sequenced and the subcellular localization of the protein was investigated. Pbp was both determined in the outer membrane of B. fragilis and of E. coli that expressed the cloned protein. Protease accessibility studies showed that the protein is expressed at the cell surface. Importantly, we demonstrated that Pbp is sufficient and required for plasminogen binding to whole cells in both E. coli and B. fragilis. Pbp-like proteins were also detected in some other Bacteroides subspecies. The role of this potential B. fragilis virulence factor in pathogenicity is discussed. PMID- 18403232 TI - Effective T-cell immune responses in the absence of the serine/threonine kinase RIP2. AB - The serine/threonine kinase RIP2 has been reported to be essential for Nod1 and Nod2 mediated cell activation, and has been suggested to play a role in the signaling cascade downstream of the T-cell receptor. We sought to ascertain the exact role of RIP2 in T-helper cell differentiation and CD8+ T-cell effector function in vivo and in vitro. In contrast to previous reports, we found that RIP2-deficient T cells did not exhibit impaired proliferation upon TCR engagement in vitro, and differentiation to cytokine producing Th1 or Th2 cells was normal in the absence of RIP2. These results were confirmed in vivo, as wild-type and RIP2-deficient virus-specific CD8+ T cells expanded comparably in mice after LCMV infection. Wild-type and RIP2-deficient CD4+ and CD8+ T cells from infected mice also showed similar proliferation and cytokine production when restimulated with full or partial agonist peptides ex vivo. Furthermore, no significant difference in adaptive T-cell responses could be observed between wild-type and RIP2 deficient mice after Listeria monocytogenes infection. Thus contrary to early reports, our data show that RIP2 is not an essential component of the TCR signaling machinery. PMID- 18403233 TI - An antagonist of lipid A action in mammals has complex effects on lipid A induction of defence responses in the model plant Arabidopsis thaliana. AB - Lipopolysaccharides, the ubiquitous part of the outer membrane of Gram-negative bacteria, and their derivatives are recognised by plants to trigger or potentiate particular defence responses such as induction of genes encoding pathogenesis related proteins. The molecular mechanisms of LPS perception that underpin these effects in plants are, however, unknown. Here, lipid A from Halomonas magadiensis, which is an antagonist of lipid A action in human cells, was used to investigate lipid A action in plants. Our findings offer an insight into the different structural requirements for direct induction and potentiation of plant defences by lipid A. PMID- 18403234 TI - Nasal immunization of mice with Lactobacillus casei expressing the Pneumococcal Surface Protein A: induction of antibodies, complement deposition and partial protection against Streptococcus pneumoniae challenge. AB - Strategies for the development of new vaccines against Streptococcus pneumoniae infections try to overcome problems such as serotype coverage and high costs, present in currently available vaccines. Formulations based on protein candidates that can induce protection in animal models have been pointed as good alternatives. Among them, the Pneumococcal Surface Protein A (PspA) plays an important role during systemic infection at least in part through the inhibition of complement deposition on the pneumococcal surface, a mechanism of evasion from the immune system. Antigen delivery systems based on live recombinant lactic acid bacteria (LAB) represents a promising strategy for mucosal vaccination, since they are generally regarded as safe bacteria able to elicit both systemic and mucosal immune responses. In this work, the N-terminal region of clade 1 PspA was constitutively expressed in Lactobacillus casei and the recombinant bacteria was tested as a mucosal vaccine in mice. Nasal immunization with L. casei-PspA 1 induced anti-PspA antibodies that were able to bind to pneumococcal strains carrying both clade 1 and clade 2 PspAs and to induce complement deposition on the surface of the bacteria. In addition, an increase in survival of immunized mice after a systemic challenge with a virulent pneumococcal strain was observed. PMID- 18403235 TI - Atopobium vaginae triggers an innate immune response in an in vitro model of bacterial vaginosis. AB - Bacterial vaginosis is the most common vaginal disorder among women of reproductive age. The pathogenesis of bacterial vaginosis is poorly understood, but is defined by a transition in the vaginal flora from the predominant Lactobacillus species to other bacterial species such as Atopobium vaginae and Gardnerella vaginalis. This change is associated with an increase in vaginal cytokine secretion. We hypothesize that vaginal epithelial cells respond to bacterial vaginosis-associated bacteria by triggering an innate immune response. We observed that vaginal epithelial cells secreted interleukin-6 and interleukin 8 in response to Atopobium vaginae and Gardnerella vaginalis, but not to Lactobacillus crispatus. Atopobium vaginae induced increased levels of interleukin-6 and interleukin-8 transcripts, as well as increased transcripts for the antimicrobial peptide beta-defensin 4. This innate immune response required live bacteria capable of protein synthesis in direct contact with vaginal epithelial cells. The response of vaginal epithelial cells was mediated by Toll like receptor 2, required the adaptor protein MyD88, and involved activation of the NFkappaB signaling pathway. These results suggest that Atopobium vaginae stimulates an innate immune response from vaginal epithelial cells, leading to localized cytokine and defensin production, and possibly contributes to the pathogenesis of bacterial vaginosis. PMID- 18403236 TI - Molecular and biological characterization of histidine triad protein in group A streptococci. AB - Four Streptococcus pneumoniae genes, phtA, phtB, phtD, and phtE, as well as the slr gene of group A streptococci (GAS), encode proteins with a histidine triad motif (HxxHxH). Pht proteins function as protective antigens against S. pneumoniae infection. A search of the GAS genome database identified a novel protein, HtpA, possessing five histidine triad motifs. The htpA gene was shown to encode a 92.5-kDa protein located downstream of the fbaA and lbp genes, while Western blot analyses revealed that HtpA protein was expressed on the cell surfaces of all group A, B, C, and G streptococcal isolates tested. Immunization of mice with rHtpA induced antigen-specific antibody production and was effective after a single immunization, with antibody titers remaining constant for at least 84days. In addition, HtpA-immunized mice survived after challenge with GAS strains isolated from patients with streptococcal toxic shock syndrome for significantly longer periods than sham-immunized mice. In that experiment, the HtpA-specific antibody was effectively induced by a single immunization and the specific antibody titer remained constant for at least 84days. These results indicate that the novel histidine triad protein HtpA is a candidate vaccine for GAS infection. PMID- 18403237 TI - Differential expression of iutA and ibeA in the early stages of infection by extra-intestinal pathogenic E. coli. AB - Extraintestinal pathogenic Escherichia coli strains are responsible for a number of infections in humans and animals. Several ExPEC virulence genes have already been described such as iutA involved in iron acquisition and ibeA required for invasion of eukaryotic cells. In this study we used the chicken model to study the expression of iutA and ibeA by two ExPEC strains during growth of bacteria in LB medium and during the infection. Expression of iutA and ibeA were shown to be higher in stationary phase than in exponential phase in vitro. During infection, iutA expression was increased at least 50-fold in the airsac and in the lung 3, 6 and 24h. p.i. compared to in vitro grown bacteria. Expression of ibeA was increased 2.5-9-fold in the airsac in the early stages of the infection only. This is the first report analyzing quantitatively the expression of ExPEC virulence genes during the course of the infection. The model described could be useful to study the expression of other ExPEC virulence genes. PMID- 18403238 TI - Microarray analysis of transcriptional responses to infection by herpes simplex virus types 1 and 2 and their US3-deficient mutants. AB - Herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) induce similar responses in infected cells and animals but differ in several significant respects. Previous studies have shown that defects in the US3-encoded protein kinase greatly affect both viruses in their interactions with cells and hosts. To investigate the impact of infection with HSV-1, HSV-2 and their US3-deficient mutants (DeltaUS3) on cellular transcriptional responses, we performed a global microarray analysis on human epithelial HEp-2 cells that were mock-infected, or infected with wild type (WT) HSV-1, HSV-2 and their DeltaUS3 mutants. Among 54,765 probe sets examined, only 1156 (approximately 2.1%) and 2006 (approximately 3.7%) genes increased by at least fourfold at 9h postinfection in WT HSV-1 and HSV-2-infected cells, respectively. Unexpectedly, HSV-2 infection increases mRNA levels for a larger number of cellular genes than HSV-1 infection. Additionally, DeltaUS3 infection upregulated the expression of a larger number of cellular genes than WT infection. The genes affected by HSV infection were assigned to various groups of functional classes and cellular pathways. We have thus identified cellular genes whose expression was similarly or differently changed by infection with each virus. PMID- 18403239 TI - Could inhaled mite faeces introduce pathogens to the lungs? PMID- 18403240 TI - Characterisation of Escherichia coli strains involved in transcytosis across gut epithelial cells exposed to metabolic and inflammatory stress. AB - Translocation of normally non-pathogenic bacteria across the gut may drive inflammatory responses associated with sepsis and inflammatory bowel disease. Recent evidence suggests translocation may not be purely passive, but occurs via novel transcellular pathways activated in enterocytes by inflammatory and metabolic stress. The specificity of this pathway with respect to different E. coli strains and other bacterial species, and possible molecular determinants of the "translocating" phenotype have been investigated. Translocation of E. coli strains and other bacteria was studied across Caco-2 monolayers exposed to different forms of cellular stress. All bacteria, apart from the pathogen Shigella sonnei, exhibited low levels of translocation in untreated monolayers. However, following enterocyte stress, translocation of E. coli strains C25 and HBTEC-1 was markedly stimulated, accompanied by increased internalisation into enterocytes. C25 and HBTEC-1 were typed to ECOR group A and group D respectively. Pathoarray analysis showed both strains had profiles quite different to those predicted for typical ExPEC isolates, lacking many of the genes associated with pathogenicity, although they contained several ORFs in common with ExPEC isolates. These data suggest translocating E. coli strains associated with infections are not opportunistic ExPEC strains but may comprise a separate group of E. coli strains. PMID- 18403241 TI - Functional analysis of the multi-copper oxidase from Legionella pneumophila. AB - Multicopper oxidases have been described to have functions in copper tolerance, manganese oxidation, and iron oxidation in a range of bacteria. The putative cytoplasmic membrane multicopper oxidase from Legionella pneumophila was investigated. The mcoL gene was found to be critical for aerobic extracellular growth under either iron-limiting conditions or in the presence of ferrous Fe(II) iron, as a sole source of this essential metal. The mcoL mutants showed minor growth defects when grown in the presence of Fe(III) as the iron source. In contrast, intracellular growth and survival was not affected by the absence of the mcoL gene regardless of available iron concentration. The evidence presented here could indicate a possible role for mcoL in prevention of the toxic effects of ferrous iron during aerobic conditions. However, a function in high-affinity acquisition of iron could also be possible given the inability of the McoL mutants to grow aerobically under iron-limiting conditions. PMID- 18403242 TI - Interaction of epithelial cell membrane rafts with Paracoccidioides brasiliensis leads to fungal adhesion and Src-family kinase activation. AB - Membrane rafts are cholesterol- and sphingolipid-enriched cell membrane domains, which are ubiquitous in mammals and play an essential role in different cellular functions, including host cell-pathogen interaction. In this work, by using several approaches, we demonstrated the involvement of epithelial cell membrane rafts in adhesion process of the pathogenic fungus Paracoccidioides brasiliensis. This conclusion was supported by the localization of ganglioside GM1, a membrane raft marker, at P. brasiliensis-epithelial cell contact sites, and by the inhibition of this fungus adhesion to host cells pre-treated with cholesterol extractor (methyl-beta-cyclodextrin, MbetaCD) or -binding (nystatin) agents. In addition, at a very early stage of P. brasiliensis-A549 cell interaction, this fungus promoted activation of Src-family kinases (SFKs) and extracellular signal regulated kinase 1/2 (ERK1/2) of these epithelial cells. Whereas SFKs were partially responsible for activation of ERK1/2, membrane raft disruption with MbetaCD in A549 cells led to total inhibition of SFK activation. Taking together, these data indicate for the first time that epithelial cell membrane rafts are essential for P. brasiliensis adhesion and activation of cell signaling molecules. PMID- 18403243 TI - The essential role of IFN-gamma in the control of lethal Aggregatibacter actinomycetemcomitans infection in mice. AB - Inflammatory immune reactions in response to periodontopathogens trigger periodontal destruction, but their role to protect the host against infection remains unknown. Thus, we examined the mechanisms by which IFN-gamma modulates the outcome of Aggregatibacter actinomycetemcomitans-induced periodontal disease in mice. Our results showed that IFN-gamma deficient mice developed less severe periodontitis in response to A. actinomycetemcomitans infection, characterized by significant lower alveolar bone loss and inflammatory reaction. However, the absence of IFN-gamma results in increased bacterial load in periodontal tissues and higher acute phase reaction, followed by a disseminated bacterial infection and mice death during the course of the disease. Such impaired host response was found to be associated with a reduction in the levels of inflammatory cytokines and chemokines and in the number of GR1+, F4/80+, CD4+ and CD8+ leukocytes in the diseased periodontium of IFN-gamma deficient mice. In addition, the levels of both antimicrobial mediators myeloperoxidase and inducible nitric oxide synthase were also found to be reduced in IFN-KO mice. Our results demonstrate for the first time that a periodontal infection may be lethal in an immunocompromised host. In addition, the mechanisms involved in IFN-gamma mediated cell migration to diseased periodontal tissues, and its essential role to control A. actinomycetemcomitans infection were clarified. PMID- 18403244 TI - Enhanced proinflammatory response to the Candida albicans gpi7 null mutant by murine cells. AB - The Candida albicans gpi7/gpi7 null mutant strain (Deltagpi7), which is affected in glycosylphosphatidylinositol (GPI) anchor biosynthesis, showed a reduced virulence following systemic infection of C57BL/6 mice. In vitro production of TNF-alpha, IL-6 and IL-1beta by macrophages in response to Deltagpi7 cells was significantly increased as compared to control (wild type GPI7/GPI7 and revertant gpi7/GPI7) cells; this probably contributes to the enhanced recruitment of neutrophils to the peritoneal cavity in response to Deltagpi7 cells. Survival of knockout mice for Toll-like receptor (TLR) 2 and TLR4 following intravenous injection of Deltagpi7 cells showed no significant differences as compared to C57BL/6 mice. In vitro production of TNF-alpha by macrophages and neutrophil recruitment were significantly inhibited in TLR2-/- mice in response to control yeast strains. Interestingly both TNF-alpha production and neutrophil recruitment in response to Deltagpi7 were significantly increased in all three types of mice, with no differences among them, and laminarin failed to inhibit this increased production of TNF-alpha. These results indicate that the enhanced proinflammatory response to Deltagpi7 does not involve recognition through TLR2, TLR4 nor dectin 1. Therefore, complete GPI anchors confer surface properties that are involved in modulation of cytokine production by macrophages in response to C. albicans. PMID- 18403245 TI - Mean platelet volume (MPV) as an inflammatory marker in ankylosing spondylitis and rheumatoid arthritis. AB - AIMS: The aim of this retrospective study was to investigate the correlation between MPV and the clinical disease activity indices of rheumatoid arthritis and ankylosing spondylitis. METHODS: The study consisted of 32 active RA patients (males/females: 7/25, mean age: 49+/-13) and 30 active AS patients (males/females: 15/15, mean age: 36+/-12) along with 26 osteoarthritis (OA) patients (males/females: 4/22, mean age: 52+/-8) and 29 age-matched healthy subjects (males/females: 5/24, mean age: 41+/-7) as control groups for RA and AS, respectively. RESULTS: MPV was significantly lower in both AS patients and RA patients with active disease as compared to controls (RA vs OA p<0.001, AS vs healthy subjects p<0.001). After treatment MPV values significantly increased in AS and RA (p<0.001 for all). However, MPV values remained somewhat lower in RA patients than OA patients (p=0.019). There was a negative correlation between MPV values and BASDAI scores in AS patients after two months of treatment (r=-0.507; p=0.004). CONCLUSION: Our results suggest that assessment of MPV may provide additional information about inflammation in AS and RA. PMID- 18403246 TI - Vascular pathology in patients with idiopathic Parkinson's disease. AB - To study the impact of brain vessel pathology on the clinical status of Parkinson's disease (PD), in 57 consecutive patients the clinical and neuropsychological data were compared with clinical MRI signs of vascular impairment and with the ultrasound brain vessel investigations. There was a significant correlation between clinical and cognitive status and intimomedial thickness, which is an indicator of large vessel impairment. Cognitive status was significantly related to the pulsatility index (an indicator of small vessel impairment). This study provides evidence that subclinical vascular pathology could influence the clinical status by contributing to motor and cognitive dysfunction in PD. PMID- 18403247 TI - Anticancer effects of ginsenoside Rg1, cinnamic acid, and tanshinone IIA in osteosarcoma MG-63 cells: nuclear matrix downregulation and cytoplasmic trafficking of nucleophosmin. AB - Ginsenoside Rg1, cinnamic acid, and tanshinone IIA are effective anticancer and antioxidant constituents of traditional Chinese herbal medicines of Ginseng (Panax ginseng), Xuanshen (Radix scrophulariae), and Danshen (Salvia mitiorrhiza), respectively. There was insufficient study on molecular mechanisms of anticancer effects of those constituents and their targets were unknown. We chose nucleophosmin as a candidate molecular target because it is frequently mutated and upregulated in various cancer cells. Nucleophosmin is a major nucleolus phosphoprotein that involves in rRNA synthesis, maintaining genomic stability, and normal cell division and its haploinsufficiency makes cell more susceptible to oncogenic assault. Ginsenoside Rg1, cinnamic acid, and tanshinone IIA treatment of osteosarcoma MG-63 cells decreased nucleophosmin expression in nuclear matrix and induced nucleophosmin translocation from nucleolus to nucleoplasm and cytoplasm, a process of dedifferentiating transformed cells. Using immunogold electro-microscopy, we found at the first time that nucleophosmin was localized on nuclear matrix intermediate filaments that had undergone restorational changes after the treatments. Nucleophosmin also functions as a molecular chaperone that might interact with multiple oncogenes and tumor suppressor genes. We found that oncogenes c-myc, c-fos and tumor suppressor genes, P53, Rb were regulated by ginsenoside Rg1, cinnamic acid, and tanshinone IIA as well. In present study, we identified nucleophosmin as a molecular target of the effective anticancer constituents of t Ginseng, Xuanseng, and Danseng that down-regulated nucleophosmin in nuclear matrix, changed its trafficking from nucleolus to cytoplasm, and regulated several oncogenes and tumor suppressor genes. Therefore, we postulate that Ginsenoside Rg1, cinnamic acid, and tanshinone IIA could serve as protective agents in cancer prevention and treatment. PMID- 18403248 TI - Role of an invariant lysine residue in folate binding on Escherichia coli thymidylate synthase: calorimetric and crystallographic analysis of the K48Q mutant. AB - Thymidylate synthase (TS) catalyzes the reductive methylation of deoxyuridine monophosphate (dUMP) using methylene tetrahydrofolate (CH(2)THF) as cofactor, the glutamate tail of which forms a water-mediated hydrogen bond with an invariant lysine residue of this enzyme. To understand the role of this interaction, we studied the K48Q mutant of Escherichia coli TS using structural and biophysical methods. The k(cat) of the K48Q mutant was 430-fold lower than wild-type TS in activity, while the K(m) for the (R)-stereoisomer of CH(2)THF was 300 microM, about 30-fold larger than K(m) from the wild-type TS. Affinity constants were determined using isothermal titration calorimetry, which showed that binding was reduced by one order of magnitude for folate-like TS inhibitors, such as propargyl-dideazafolate (PDDF) or compounds that distort the TS active site like BW1843U89 (U89). The crystal structure of the K48Q-dUMP complex revealed that dUMP binding is not impaired in the mutant, and that U89 in a ternary complex of K48Q-nucleotide-U89 was bound in the active site with subtle differences relative to comparable wild-type complexes. PDDF failed to form ternary complexes with K48Q and dUMP. Thermodynamic data correlated with the structural determinations, since PDDF binding was dominated by enthalpic effects while U89 had an important entropic component. In conclusion, K48 is critical for catalysis since it leads to a productive CH(2)THF binding, while mutation at this residue does not affect much the binding of inhibitors that do not make contact with this group. PMID- 18403249 TI - c-Cbl facilitates cytoskeletal effects in v-Abl transformed fibroblast through Rac1- and Rap1-mediated signaling. AB - c-Cbl functions as a multifunctional adaptor and an E3 ubiquitin ligase. Several studies have shown that c-Cbl is involved in cytoskeleton-mediated events, but the molecular mechanisms linking c-Cbl to cytoskeletal rearrangements remain to be elucidated. Our previous results indicated that c-Cbl facilitates spreading and migration of v-Abl-transformed NIH 3T3 fibroblasts and suggested that small GTPases play important roles in the cytoskeletal effects of c-Cbl in this system. To elucidate the individual contributions of small GTPases to these effects, we assessed the roles of endogenous Rac1, RhoA and Rap1 in the c-Cbl-dependent spreading and migration of v-Abl-transformed fibroblasts overexpressing c-Cbl, using RNAi. Furthermore, since it has been shown that Rap1 can act as an upstream regulator of Rac1 in inducing cell spreading, we analyzed the interplay between Rap1 and Rac1 in the signaling pathways connecting c-Cbl to the cytoskeletal events. Our results indicate that Rac1 is essential for cell migration and spreading, whereas activation of RhoA exerts a negative effect. We have also shown that Rap1 is essential for cell spreading, although not for migration in our experimental system. Furthermore, we provide evidence that Rap1 is located upstream of Rac1 in one of the signaling pathways that regulate c-Cbl-facilitated cell spreading. Overall, our findings are consistent with the model describing the connection of c-Cbl to the cytoskeletal rearrangements via two pathways, one of which is mediated by PI3K and Rac1, and the other, by CrkL/C3G, Rap1 and Rac1. PMID- 18403250 TI - Matrix metalloproteinase-2 mediates stretch-induced activation of skeletal muscle satellite cells in a nitric oxide-dependent manner. AB - When skeletal muscle is stretched or injured, myogenic satellite cells are activated to enter the cell cycle. This process depends on nitric oxide (NO) production, release of hepatocyte growth factor (HGF) from the extracellular matrix, and presentation of HGF to the c-met receptor. Matrix metalloproteinases (MMPs), a large family of zinc-dependent endopeptidases, mediate HGF release from the matrix and this step in the pathway is downstream from NO synthesis [Yamada, M., Tatsumi, R., Kikuiri, T., Okamoto, S., Nonoshita, S., Mizunoya, W., et al. (2006). Matrix metalloproteinases are involved in mechanical stretch-induced activation of skeletal muscle satellite cells. Muscle Nerve, 34, 313-319]. Experiments reported herein provide evidence that MMP2 may be involved in the NO dependent release of HGF in vitro. Whole lysate analyses of satellite cells demonstrated the presence of MMP2 mRNA and the protein. When rat satellite cells were treated with 30 microM sodium nitroprusside a NO donor or mechanical cyclic stretch for 2h period, inactive proMMP2 (72 kDa) was converted into 52-kDa form and this processing was abolished by adding a NO synthase inhibitor l-NAME (10 microM) to the stretch culture. The 52-kDa species was also generated by treatment of the recombinant MMP2 protein with 1 microM NOC-7 that can spontaneously release NO under physiological conditions without any cofactor, and its activating activity was demonstrated by applying the NOC-7-treated MMP2 to satellite cell culture. HGF release was detected in NOC-7-MMP2-conditioned media by western blotting; very little HGF was found in media that were generated from cultures receiving NOC-7-treated MMP2 (10 ng/ml) plus 250 ng/ml tissue inhibitor 1 of metalloproteinases. Therefore, results from these experiments provide evidence that NO-activated MMP2 may cause release of HGF from the extracellular matrix of satellite cells and contribute to satellite cell activation. PMID- 18403251 TI - What antipriming reveals about priming. AB - Across many domains of cognition, information processing is more effective when it has been performed recently than when it has not. This effect--termed repetition priming--is ubiquitous; however, its causes are not well understood. Here, I draw attention to the concept of antipriming and argue that it is crucial for understanding priming. Antipriming is a measurable impairment in processing information owing to recent processing of other information when the representations of information overlap and compete. Strengthening one representation after its usage causes priming for that item but also antipriming for some other, non-repeated items. Recent evidence demonstrates priming and antipriming within visual object identification systems. These findings might reflect a form of maintenance relearning of superimposed knowledge representations. PMID- 18403252 TI - Cognitive control, hierarchy, and the rostro-caudal organization of the frontal lobes. AB - Cognitive control supports flexible behavior by selecting actions that are consistent with our goals and appropriate for our environment. The prefrontal cortex (PFC) has an established role in cognitive control, and research on the functional organization of PFC promises to contribute to our understanding of the architecture of control. A recently popular hypothesis is that the rostro-caudal axis of PFC supports a control hierarchy whereby posterior-to-anterior PFC mediates progressively abstract, higher-order control. This review discusses evidence for a rostro-caudal gradient of function in PFC and the theories proposed to account for these results, including domain generality in working memory, relational complexity, the temporal organization of behavior and abstract representational hierarchy. Distinctions among these frameworks are considered as a basis for future research. PMID- 18403253 TI - Chromatin-remodelling mechanisms in cancer. AB - Chromatin-remodelling mechanisms include DNA methylation, histone-tail acetylation, poly-ADP-ribosylation, and ATP-dependent chromatin-remodelling processes. Some epigenetic modifications among others have been observed in cancer cells, namely (1) local DNA hypermethylation and global hypomethylation, (2) alteration in histone acetylation/deacetylation balance, (3) increased or decreased poly-ADP-ribosylation, and (4) failures in ATP-dependent chromatin remodelling mechanisms. Moreover, these alterations can influence the response to classical anti-tumour treatments. Drugs targeting epigenetic alterations are under development. Currently, DNA methylation and histone deacetylase inhibitors are in use in cancer therapy, and poly-ADP-ribosylation inhibitors are undergoing clinical trials. Epigenetic therapy is gaining in importance in pharmacology as a new tool to improve anti-cancer therapies. PMID- 18403254 TI - Laser produced spectrum of Si(2) molecule in the region of 540-1010 nm. AB - The laser produced spectrum of Si(2) molecule is recorded for the first time using laser ablation technique in the region of 540-1010 nm. About 110 bands are observed in the entire spectral region and all these bands are classified into three band systems, viz. E-X, F-X and G-X of Si(2) molecule lying in the region of 814-1010 nm, 630-900 nm and 546-710 nm, respectively. All these electronic transitions take place from ground state X(3)Sigma(g)(-) state. The molecular constants of all these states have been determined. PMID- 18403255 TI - Ligand-field treatment for the optical and electron paramagnetic resonance spectra of ZnO:V(3+). AB - In this work, a full ligand-field energy matrix (45 x 45) diagonalization treatment for 3d(2) ions in trigonal symmetry C(3v) is performed on the basis of a central metal ion-ligand covalency model including the ligand spin-orbit (SO) coupling. Optical fine structure and electron paramagnetic resonance (EPR) spectra of ZnO:V(3+) are uniformly explained. PMID- 18403256 TI - Role of antiviral prophylaxis in renal transplant recipients at high risk of cytomegalovirus infection. PMID- 18403257 TI - Calu-3/A-549 mixed cells as a replacement for primary rhesus monkey kidney cells for virus detection. AB - BACKGROUND: Primary kidney cells derived from rhesus macaques (pRhMK) are heavily relied upon for the detection and culture of a wide range of clinically relevant viruses. The use of these cultures is problematic due to the possible presence of endogenous viruses, the need to sacrifice a primate, and the inherent variance found in primary cultures. OBJECTIVE: To develop a continuous cell line or mixed cell co-culture that could replace dependence on pRhMK cells. STUDY DESIGN: Cells from the Calu-3 and A-549 cell lines were used to prepare mixed cell monolayers that were compared to pRhMK cells for their ability to detect respiratory viruses, measles, mumps, enteroviruses, and herpes viruses. Clinically derived and laboratory virus strains were used for these comparisons in culture plates or 16 mm tubes. RESULTS: Calu-3/A-549 cells are more sensitive than pRhMK for the detection of adenovirus, enteroviruses and herpes simplex virus and are about equally sensitive for the detection of other respiratory viruses, measles, mumps and varicella-zoster virus. CONCLUSIONS: Calu-3/A-549 cells are an equivalent or better alternative to pRhMK cells for the detection of many clinically relevant viruses. PMID- 18403258 TI - First detection of enterovirus 71 from an acute flaccid paralysis case with residual paralysis in Iran. AB - In an attempt to determine the types of non-polio enteroviruses (NPEVs) in acute flaccid paralysis (AFP) cases in Iran, we detected enterovirus 71 (EV71) in an AFP case with residual paralysis for the first time. Cell culture detected no enteroviruses, while RT-PCR and subsequent sequencing revealed that the specimen was positive for EV71. EV71 is the causative agent of a variety of diseases from hand, foot and mouth disease to severe neurological complications and is now considered as an important cause of childhood acute flaccid paralysis. PMID- 18403259 TI - Prevalence and risk factors for chemical sensitivity and sensory hyperreactivity in teenagers. AB - The prevalence of chemical sensitivity (CS) and sensory hyperreactivity (SHR) was assessed in a teenage population. Among a random sample of 401 teenagers, 326 (81.3%) answered questionnaires assessing sensitivity to chemicals and noise, anxiety and depression. A subgroup of 85 teenagers conducted a capsaicin inhalation test. The estimated prevalence was 15.6% for general self-reported CS, 3.7% for CS with affective and behavioral consequences, about 1% for SHR. Sensitivity variables were positively intercorrelated. Risk factors for general CS were noise sensitivity (OR: 2.1), probable anxiety (OR: 2.5) and female sex (OR: 2.0). CS problems seem to be present also in teenagers, although less so than in adults. Furthermore, CS seems to be related to other environmental sensitivities. PMID- 18403260 TI - Domiciliary oxygen therapy improves sub-maximal exercise capacity and quality of life in chronic heart failure. AB - INTRODUCTION: A high prevalence of nocturnal hypoxia is noted in patients with chronic heart failure (CHF). Nocturnal hypoxia can be reversed by nasal oxygen or non-invasive assisted ventilation. However, controversy exists over the use of oxygen in CHF. We studied the effects of nocturnal nasal oxygen in CHF to demonstrate its effects on 6-min walk test, quality of life (QOL), NTproBNP, and echocardiographic parameters. METHODOLOGY: Ten patients aged 70+/-9 years received domiciliary oxygen (4 l/min) for one month. Oxygen was administered for a minimum of 8h every night via nasal prongs. RESULTS: A 17% improvement was seen in the 6-min walk test, 298+/-98 m to 351+/-100 m (p=0.005) and a 27% improvement in the QOL, which improved from 26+/-12 to 19+/-7 (p=0.017). Acquired echocardiographic measures including ejection fraction, pulmonary pressure and diastolic parameters did not change. CONCLUSION: Nocturnal nasal oxygen has a significant impact on sub-maximal exercise capacity and QOL in CHF. Although a previous study has demonstrated harmful haemodynamic effects of acute oxygen administration in CHF; we did not detect any change in echocardiographic parameters using current two-dimensional imaging and Doppler studies. The lack of improvement in cardiac parameters suggests a peripheral mode of action. PMID- 18403261 TI - Evaluation of the extracardiac conduit modification of the Fontan operation for thrombus formation using magnetic resonance imaging. AB - BACKGROUND: The prevalence of thrombosis after the Fontan procedure depends upon the surgical technique used and the method of detection employed. Current investigations for thrombosis lack sensitivity and specificity or, in the paediatric population, require a general anaesthetic. We undertook a study to examine the feasibility of using magnetic resonance imaging (MRI) to detect thrombosis within the conduit, cardiac chambers and pulmonary arteries after the extracardiac conduit modification of the Fontan procedure. METHODS: Of the 50 children who had undergone this procedure at our institution between 1997 and 2002, 26 were eligible for, and 13 underwent, MRI study. The mean age was 10.2 years (range 8.2-16.8 years, median 9.5 years) and the average time from operation was 63 months (range 29-79 months, median 68 months). The mean age at Fontan operation was 4.9 years (range 2.1-10.5 years). Ten were on low dose aspirin, two were on warfarin and one was not anti-coagulated. In all cases, satisfactory imaging of the venous pathways and pulmonary arteries was obtained and there were no thrombi detected. CONCLUSIONS: We conclude that MRI is a potentially useful tool for the detection of thrombus in patients who have undergone the Fontan operation. PMID- 18403262 TI - Ultrastructure and functional morphology of glandular setae and distal claws of cephalic appendages of Speleonectes tanumekes (Crustacea: Remipedia). AB - Terminal pores on crustacean setae are commonly associated with chemoreception. In this study we present an exception to that association with the description of glandular setae on maxillulary and maxillary endites of the remipede Speleonectes tanumekes. This introduces a function associated with crustacean setae beyond the general functions currently assigned to crustacean setae: sensory functions, mechanical functions, or a combination of these two. Even though the functions of the secretory products are unclear, we suggest means by which these may contribute to feeding behaviors. In addition, we describe glandular features of maxillary and maxillipedal distal claws of the same remipede species. Glandular setae and distal claws appear to share several morphological homologies. PMID- 18403264 TI - Cell- and peptide-based immunotherapeutic approaches for glioma. AB - Glioblastoma multiforme (GBM) is the most common and lethal primary malignant brain tumor. Although considerable progress has been made in surgical and radiation treatment for glioma patients, the impact of these advances on clinical outcome has been disappointing. Therefore, the development of novel therapeutic approaches is essential. Recent reports demonstrate that systemic immunotherapy using dendritic cells (DCs) or peptide vaccines is capable of inducing an antiglioma response. These approaches successfully induce an antitumor immune response and prolong survival in patients with glioma without major side effects. There are several types of glioma, so to achieve effective therapy, it might be necessary to evaluate the molecular genetic abnormalities in individual patient tumors and design novel immunotherapeutic strategies based on the pharmacogenomic findings. Here, we review recent advances in DC- and peptide-based immunotherapy approaches for patients with gliomas. PMID- 18403265 TI - CD38 at the junction between prognostic marker and therapeutic target. AB - CD38 is an ectoenzyme involved in transmembrane signaling and cell adhesion and is used as a disease marker for leukemias and myeloma. CD38 is a dependable negative prognostic marker for chronic lymphocytic leukemia (CLL). Recent evidence indicates that CD38 is a component of a complex network delivering growth and survival signals to CLL cells. In conjunction with chemokines and their receptors, CD38 also influences cell migratory responses. These considerations are the rationale for devising a CLL therapy that uses CD38 as the target. The use of reagents specifically blocking the molecule might provide a new approach for interfering with deleterious growth circuits, therefore increasing the susceptibility of leukemic cells to conventional chemotherapy. PMID- 18403266 TI - Decision trees on the web - a parasite compendium. PMID- 18403263 TI - OutFOXOing disease and disability: the therapeutic potential of targeting FoxO proteins. AB - Forkhead transcription factors have a 'winged helix' domain and regulate processes that range from cell longevity to cell death. Of the mammalian forkhead family members in the O class, FoxO1, FoxO3a and FoxO4 can fill a crucial void for the treatment of disorders that include aging, cancer, diabetes, infertility, neurodegeneration and immune system dysfunction. Yet, observations that forkhead family members also can compromise clinical utility have fueled controversy and highlight the necessity to further outline the integrated cellular pathways governed by these transcription factors. Here we discuss recent advances that have elucidated the unique cellular pathways and clinical potential of targeting FoxO proteins to develop novel therapeutic strategies and avert potential pitfalls that might be closely intertwined with its benefits for patient care. PMID- 18403267 TI - Plasmodium vivax in India. AB - Four Plasmodium species cause malaria in humans: Plasmodium vivax is the most widespread and results in pronounced morbidity. India (population >1 billion) is a major contributor to the burden of vivax malaria. With a resurgence in interest concerning the neglected burden of vivax malaria and the completion of the P. vivax genome, it is timely to review what is known concerning P. vivax in India. The P. vivax population is highly diverse in terms of relapse patterns, drug response and clinical profiles, and highly genetically variable according to studies of antigen genes, isoenzyme markers and microsatellites. The unique epidemiology of malaria in India, where P. vivax predominates over Plasmodium falciparum, renders this location ideal for studying the dynamics of co infection. PMID- 18403268 TI - What do unicellular organisms teach us about DNA methylation? AB - DNA methylation is an epigenetic hallmark that has been studied intensively in mammals and plants. However, knowledge of this phenomenon in unicellular organisms is scanty. Examining epigenetic regulation, and more specifically DNA methylation, in these organisms represents a unique opportunity to better understand their biology. The determination of their methylation status is often complicated by the presence of several differentiation stages in their life cycle. This article focuses on some recent advances that have revealed the unexpected nature of the epigenetic determinants present in protozoa. The role of the enigmatic DNA methyltransferase Dnmt2 in unicellular organisms is discussed. PMID- 18403269 TI - Clinical guidelines: some considerations. PMID- 18403270 TI - Motive for nonmedical use of prescription pain relievers in the national survey on drug use and health. PMID- 18403271 TI - Treatment of patients with complex regional pain syndrome type I with mannitol: a prospective, randomized, placebo-controlled, double-blinded study. AB - To assess the effects of intravenous administration of the free radical scavenger mannitol 10% on complaints associated with complex regional pain syndrome Type I (CRPS I), a randomized, placebo-controlled, double-blinded trial was performed. Forty-one CRPS I patients according to the Bruehl et al diagnostic criteria, were included in 2 outpatient pain clinics of 2 university medical centers and randomly assigned to receive either 10% mannitol iv in 1 L 0.9% NaCL in 4 hours for 5 consecutive days or equal volumes of 0.9% NaCL (placebo). Patients in both groups received physical therapy according to protocol and rescue pain medication if required. Complaints on impairment and disability level and quality of life were assessed up to 9 weeks after baseline, with primary measurement points at 2, 6, and 9 weeks. Monitoring of pain using the visual analogue scale took place continuously during the course of the trial. Except for a significant improvement on a subscale of the Jebsen-Taylor hand function test, no significant differences were found between mannitol and placebo treatment. Changes in both groups in the course of the trial were small and clinically irrelevant on all measurement indices. We conclude that intravenous administration of 10% mannitol is not more effective than placebo in reducing complaints for CRPS I patients and provides no addition to already-established interventions for CRPS I. Whether 10% mannitol can provide beneficial effects for subgroups of CRPS I patients with a pathophysiological profile more closely fitting the presumed mode of action for this intervention remains to be established. PERSPECTIVE: This article addresses the efficacy of the intravenous administration of the free radical scavenger mannitol for treatment of CRPS type 1. This intervention is not more effective than placebo in reducing complaints for CRPS I patients and provides no addition to already-established interventions for CRPS I. PMID- 18403273 TI - Metallothionein genes from hydrothermal crabs (Bythograeidae, Decapoda): characterization, sequence analysis, gene expression and comparison with coastal crabs. AB - Hydrothermal vent conditions can alter DNA and hydrothermal organisms may develop detoxification mechanisms and/or genetic adaptations. Hydrothermal vent animals notably synthesize a high quantity of metallothioneins (MT). Recent studies have revealed that the levels of MT within hydrothermal crustacean tissues are higher than those found in other vent animals. To improve our understanding of the environmental impacts exerted on the vent organisms, we characterized the metallothioneins (cDNA and Mt genes) of several members of the Bythograeidae (Bythograea thermydron, Cyanagraea praedator and Segonzacia mesatlantica) which is the only endemic hydrothermal crab family. In comparison, the isolation of metallothionein cDNA was also carried out in several coastal crab families. The results showed that the hydrothermal crabs possess Mt composed of three exons and two introns presenting conserved splicing signals. The cDNA sequences isolated from distinct crabs showed multiple substitutions. In spite of the unique environmental conditions, the protein sequence analysis revealed no specific amino acid residue for the MT of the three hydrothermal crabs. However, gene expression analysis performed by real-time PCR based on S. mesatlantica (hydrothermal crab) compared to Pachygrapsus marmoratus (coastal crab) confirmed the higher metallothionein induction in hydrothermal crabs suggested by others authors. PMID- 18403272 TI - A randomized, placebo-controlled, crossover trial of cannabis cigarettes in neuropathic pain. AB - The Food and Drug Administration (FDA), Substance Abuse and Mental Health Services Administration (SAMHSA), and the National Institute for Drug Abuse (NIDA) report that no sound scientific studies support the medicinal use of cannabis. Despite this lack of scientific validation, many patients routinely use "medical marijuana," and in many cases this use is for pain related to nerve injury. We conducted a double-blinded, placebo-controlled, crossover study evaluating the analgesic efficacy of smoking cannabis for neuropathic pain. Thirty-eight patients with central and peripheral neuropathic pain underwent a standardized procedure for smoking either high-dose (7%), low-dose (3.5%), or placebo cannabis. In addition to the primary outcome of pain intensity, secondary outcome measures included evoked pain using heat-pain threshold, sensitivity to light touch, psychoactive side effects, and neuropsychological performance. A mixed linear model demonstrated an analgesic response to smoking cannabis. No effect on evoked pain was seen. Psychoactive effects were minimal and well tolerated, with some acute cognitive effects, particularly with memory, at higher doses. PERSPECTIVE: This study adds to a growing body of evidence that cannabis may be effective at ameliorating neuropathic pain, and may be an alternative for patients who do not respond to, or cannot tolerate, other drugs. However, the use of marijuana as medicine may be limited by its method of administration (smoking) and modest acute cognitive effects, particularly at higher doses. PMID- 18403274 TI - Cellular and molecular mechanisms underlying plasticity in uterine sympathetic nerves. AB - Dynamic responses of uterine sympathetic nerves to changes in the circulating levels of sex hormones represent one of the most remarkable examples of physiological plasticity in the adult autonomic nervous system. The density of uterine sympathetic nerves is markedly and irreversibly reduced following puberty, and shows phases of degeneration and regeneration during the natural oestrous cycle. Even more remarkable, uterine sympathetic nerves degenerate during normal pregnancy and regenerate following delivery. Plasticity in uterine sympathetic nerves was initially interpreted as a selective effect of sex hormones on the system of paracervical short adrenergic neurons supplying the uterus. In the last decade, the alternative explanation that sex hormones might alter the ability of the uterine tissue to support its innervation began to be explored and current evidence indicates that oestrogen and pregnancy elicit changes in the neuritogenic properties of the target uterine tissue. In addition, there are indications that sex hormones may also affect the receptivity of uterine-related sympathetic neurons to target-derived signals. Although the nature of these signals is still fragmentary, there is evidence for the contribution of a range of molecules, including neurotrophins, pro-neurotrophins and chemorepulsive signals of the semaphorin family. This review summarizes some general features of plasticity in uterine sympathetic nerves and highlights recent investigations of the cellular and molecular mechanisms underlying this dramatic model of natural plasticity. PMID- 18403275 TI - Gastric endoscopy in the 21st century: appropriate use of an invasive procedure in the era of non-invasive testing. AB - BACKGROUND: The acceptance of the premise that Helicobacter pylori infection is aetiologically related to gastric cancer and peptic ulcer and that the risk of gastric cancer among Helicobacter pylori infected individuals is related to the extent, severity and duration of atrophic gastritis has led to major changes in medical and endoscopic practices. The development of non-invasive methods to detect Helicobacter pylori and to estimate the extent and severity of gastritis has reduced the need for diagnostic endoscopy in asymptomatic individuals. METHODS AND RESULTS: Here we provide recommendations regarding deciding whether non-invasive and endoscopic assessment of the gastric mucosa is preferred. We also include specific recommendations and caveats regarding the preferred biopsy number and sites as well as the identification of specimens, to allow the pathologist to reliable stage the severity and extent of gastritis, and thus provide prognostic information needed for patient managements (e.g., whether endoscopic surveillance is recommended). CONCLUSION: In summary, while there is clearly a role for gastric endoscopy and endoscopic biopsy in the Helicobacter pylori era, obtaining useful diagnostic and prognostic information is critically dependent upon attention to detail with regard to biopsy site and identification as to the location from where the specimen was taken. PMID- 18403276 TI - Breast enlargement after thoracoscopic sympathectomy. PMID- 18403277 TI - Online access to trends in endocrinology and metabolism. PMID- 18403278 TI - Complex Complexes: Report of an NIDDK Workshop on Coactivators and Corepressors, Bethesda, USA, 15-16 December 1998. AB - Exciting new observations about receptor-interacting and nuclear accessory proteins and their ability to regulate gene expression have provided insight into hormonal regulation of gene expression. The goal of a recent NIDDK workshop was to discuss how diverse signaling pathways could converge on target genes and provide regulatory input to either activate or repress transcription. PMID- 18403279 TI - The 1999 Neuroendocrine Workshop on Food Intake, Energy Metabolism and Obesity, San Diego, CA, USA, 9-11 June 1999. AB - The recent American Endocrine Society Annual Workshop was attended by over 125 students, senior investigators and Fellows. The workshop focused on the latest developments in central nervous system (CNS) mechanisms (mainly peptidergic) that influence food intake and energy expenditure, and the biological consequences of altered energy states on reproduction. PMID- 18403280 TI - An adaptive P300-based online brain-computer interface. AB - The P300 component of an event related potential is widely used in conjunction with brain-computer interfaces (BCIs) to translate the subjects intent by mere thoughts into commands to control artificial devices. A well known application is the spelling of words while selection of the letters is carried out by focusing attention to the target letter. In this paper, we present a P300-based online BCI which reaches very competitive performance in terms of information transfer rates. In addition, we propose an online method that optimizes information transfer rates and/or accuracies. This is achieved by an algorithm which dynamically limits the number of subtrial presentations, according to the subject's current online performance in real-time. We present results of two studies based on 19 different healthy subjects in total who participated in our experiments (seven subjects in the first and 12 subjects in the second one). In the first, study peak information transfer rates up to 92 bits/min with an accuracy of 100% were achieved by one subject with a mean of 32 bits/min at about 80% accuracy. The second experiment employed a dynamic classifier which enables the user to optimize bitrates and/or accuracies by limiting the number of subtrial presentations according to the current online performance of the subject. At the fastest setting, mean information transfer rates could be improved to 50.61 bits/min (i.e., 13.13 symbols/min). The most accurate results with 87.5% accuracy showed a transfer rate of 29.35 bits/min. PMID- 18403281 TI - Local temporal common spatial patterns for robust single-trial EEG classification. AB - In this paper, we propose a novel optimal spatio-temporal filter, termed local temporal common spatial patterns (LTCSP), for robust single-trial elctroencephalogram (EEG) classification. Different from classical common spatial patterns (CSP) that uses only global spatial covariances to compute the optimal filter, LTCSP considers temporally local information in the variance modelling. The underlying manifold variances of EEG signals contain more discriminative information. LTCSP is an extension to CSP in the sense that CSP can be derived from LTCSP under a special case. By constructing an adjacency matrix, LTCSP is formulated as an eigenvalue problem. So, LTCSP is computationally as straightforward as CSP. However, LTCSP has better discrimination ability than CSP and is much more robust. Simulated experiment and real EEG classification demonstrate the effectiveness of the proposed LTCSP method. PMID- 18403282 TI - Variable patterned pudendal nerve stimuli improves reflex bladder activation. AB - We evaluated variable patterns of pudendal nerve (PN) stimuli for reflex bladder excitation. Reflex activation of the bladder has been demonstrated previously with 20-33 Hz continuous stimulation of PN afferents. Neuronal circuits accessed by afferent mediated pathways may respond better to physiological patterned stimuli than continuous stimulation. Unilateral PN nerve cuffs were placed in neurologically intact male cats. PN stimulation (0.5-100 Hz) was performed under isovolumetric conditions at bladder volumes up to the occurrence of distension evoked reflex contractions. Stimulus evoked reflex bladder contractions were elicited in eight cats. Across all experiments, bursting of 2-10 pulses at 100 200 Hz repeated at continuous stimulation frequencies evoked significantly larger bladder responses than continuous (single pulse) stimulation (52.0+/-44.5%). Bladder excitation was also effective at 1 Hz continuous stimuli, which is lower than typically reported. Variable patterned pulse bursting resulted in greater evoked reflex bladder pressures and increased the potential stimulation parameter space for effective bladder excitation. Improved bladder excitation should increase the efficacy of neuroprostheses for bladder control. PMID- 18403283 TI - SIMONE: a realistic neural network simulator to reproduce MEA-based recordings. AB - Contemporary multielectrode arrays (MEAs) used to record extracellular activity from neural tissues can deliver data at rates on the order of 100 Mbps. Such rates require efficient data compression and/or preprocessing algorithms implemented on an application specific integrated circuit (ASIC) close to the MEA. We present SIMONE (Statistical sIMulation Of Neuronal networks Engine), a versatile simulation tool whose parameters can be either fixed or defined by a probability distribution. We validated our tool by simulating data recorded from the first olfactory relay of an insect. Different key aspects make this tool suitable for testing the robustness and accuracy of neural signal processing algorithms (such as the detection, alignment, and classification of spikes). For instance, most of the parameters can be defined by a probabilistic distribution, then tens of simulations may be obtained from the same scenario. This is especially useful when validating the robustness of the processing algorithm. Moreover, the number of active cells and the exact firing activity of each one of them is perfectly known, which provides an easy way to test accuracy. PMID- 18403284 TI - A collaborative wheelchair system. AB - This paper describes a novel robotic wheelchair, and reports experiments to evaluate its efficiency and understand how human operators use it. The concept at the heart of the collaborative wheelchair assistant (CWA) is to rely on the user's motion planning skills while assisting the maneuvering with flexible path guidance. The user decides where to go and controls the speed (including start and stop), while the system guides the wheelchair along software-defined guide paths. An intuitive path editor allows the user to avoid dangers or obstacles online and to modify the guide paths at will. By using the human sensory and planning systems, no complex sensor processing or artificial decision system is needed, making the system safe, simple, and low-cost. We investigated the performance of the CWA on its interaction with able-bodied subjects and motion efficiency. The results show that path guidance drastically simplifies the control. Using the CWA, the wheelchair user needs little effort from the first trial, while moving efficiently with a conventional wheelchair requires adaptation. PMID- 18403285 TI - Psychophysical experiment of vibrotactile pattern perception by human fingertip. AB - In this paper, a preliminary experimentation is carried out to investigate the necessary stimulation conditions for a pattern display that may be used for the blind to display obstacle distribution. To design an efficient vibrotactile display device of array-type and compact size, the effect of some main parameters, such as the amplitude and frequency of vibration and the spacing and quantity of vibrated pins on vibrotactile perception, should be considered. Through experiment and analysis, the minimal amplitude of vibrating stimuli and the spacing effect on the perception of vibrotactile pattern are investigated and discussed. Also an appropriate type of actuator satisfying the requirements of a compact tactile display device for the effective transfer of pattern information is discussed. PMID- 18403286 TI - Portable preimpact fall detector with inertial sensors. AB - Falls and the resulting hip fractures in the elderly are a major health and economic problem. The goal of this study was to investigate the feasibility of a portable preimpact fall detector in detecting impending falls before the body impacts on the ground. It was hypothesized that a single sensor with the appropriate kinematics measurements and detection algorithms, located near the body center of gravity, would be able to distinguish an in-progress and unrecoverable fall from nonfalling activities. The apparatus was tested in an array of daily nonfall activities of young (n=10) and elderly (n=14) subjects, and simulated fall activities of young subjects. A threshold detection method was used with the magnitude of inertial frame vertical velocity as the main variable to separate the nonfall and fall activities. The algorithm was able to detect all fall events at least 70 ms before the impact. With the threshold adapted to each individual subject, all falls were detected successfully, and no false alarms occurred. This portable preimpact fall detection apparatus will lead to the development of a new generation inflatable hip pad for preventing fall-related hip fractures. PMID- 18403287 TI - Modeling and preliminary testing socket-residual limb interface stiffness of above-elbow prostheses. AB - The interface between the socket and residual limb can have a significant effect on the performance of a prosthesis. Specifically, knowledge of the rotational stiffness of the socket-residual limb (S-RL) interface is extremely useful in designing new prostheses and evaluating new control paradigms, as well as in comparing existing and new socket technologies. No previous studies, however, have examined the rotational stiffness of S-RL interfaces. To address this problem, a math model is compared to a more complex finite element analysis, to see if the math model sufficiently captures the main effects of S-RL interface rotational stiffness. Both of these models are then compared to preliminary empirical testing, in which a series of X-rays, called fluoroscopy, is taken to obtain the movement of the bone relative to the socket. Force data are simultaneously recorded, and the combination of force and movement data are used to calculate the empirical rotational stiffness of elbow S-RL interface. The empirical rotational stiffness values are then compared to the models, to see if values of Young's modulus obtained in other studies at localized points may be used to determine the global rotational stiffness of the S-RL interface. Findings include agreement between the models and empirical results and the ability of persons to significantly modulate the rotational stiffness of their S-RL interface a little less than one order of magnitude. The floor and ceiling of this range depend significantly on socket length and co-contraction levels, but not on residual limb diameter or bone diameter. Measured trans-humeral S-RL interface rotational stiffness values ranged from 24-140 Nm/rad for the four subjects tested in this study. PMID- 18403288 TI - Application of tripolar concentric electrodes and prefeature selection algorithm for brain-computer interface. AB - For persons with severe disabilities, a brain-computer interface (BCI) may be a viable means of communication. Lapalacian electroencephalogram (EEG) has been shown to improve classification in EEG recognition. In this work, the effectiveness of signals from tripolar concentric electrodes and disc electrodes were compared for use as a BCI. Two sets of left/right hand motor imagery EEG signals were acquired. An autoregressive (AR) model was developed for feature extraction with a Mahalanobis distance based linear classifier for classification. An exhaust selection algorithm was employed to analyze three factors before feature extraction. The factors analyzed were 1) length of data in each trial to be used, 2) start position of data, and 3) the order of the AR model. The results showed that tripolar concentric electrodes generated significantly higher classification accuracy than disc electrodes. PMID- 18403289 TI - A model of selective activation of the femoral nerve with a flat interface nerve electrode for a lower extremity neuroprosthesis. AB - Functional electrical stimulation (FES) can restore limb movements through electrically initiated, coordinated contractions of paralyzed muscles. The peripheral nerve is an attractive site for stimulation using cuff electrodes. Many applications will require the electrode to selectively activate many smaller populations of axons within a common nerve trunk. The purpose of this study is to computationally model the performance of a flat interface nerve electrode (FINE) on the proximal femoral nerve for standing and stepping applications. Simulations investigated multiple FINE configurations to determine the optimal number and locations of contacts for the maximum muscular selectivity. Realistic finite element method (FEM) models were developed from digitized cross sections from cadaver femoral nerve specimens. Electrical potentials were calculated and interpolated voltages were applied to a double-cable axon model. Model output was analyzed to determine selectivity and estimate joint moments with a musculoskeletal model. Simulations indicated that a 22-contact FINE will produce the greatest selectivity. Simulations predicted that an eight-contact FINE can be expected to selectively stimulate each of the six muscles innervated by the proximal femoral nerve, producing a sufficient knee extension moment for the sit to-stand transition and contributing 60% of the hip flexion moment needed during gait. We conclude that, whereas more contacts produce greater selectivity, eight channels are sufficient for standing and stepping with an FES system using a FINE on the common femoral nerve. PMID- 18403290 TI - Friend: a communication aid for persons with disabilities. AB - Computers offer valuable opportunities to people with physical disabilities. For example, a computer can allow someone with severe speech and motor-impairment to engage more fully with the world. This paper describes the design of a communication aid for motor-impaired users, who literally use computers as their communication partners. Currently, a low cost interface suitable for different types of motor-impaired users is hardly available. Additionally, the target audience of existing such systems is very much limited. The present work solved these problems by its adaptation mechanism. The adaptation mechanism provides an appropriate interface from an interface bank for each user before start of an interaction. The adaptation mechanism is continued during as well as after the end of interactions to make the system personalized to individual user. PMID- 18403291 TI - [Hungarian Oncology--Dear Colleagues]. PMID- 18403293 TI - [Efforts to enhance the efficiency of tumor chemotherapy]. AB - Antiproliferative cytotoxic agents, of which several are still in clinical practice nowadays, could be characterized by low selectivity, narrow therapeutic index, medium or serious side effects and rapid formation of resistance. In the limited efficacy of these drugs several factors are playing a role such as the age, gender and pharmacogenetics of the patients, the morphological and biological feature of the tumor, moreover, pharmacokinetics and pharmacodynamics of the drugs. The question could be justified if there are methods which, by influencing the above parameters, are helpful in enhancing the efficacy and decreasing the toxic side effects of these drugs. Since a long time we have been interested in evaluating methods of preclinical and clinical level for increasing drug efficacy. The aim of this minireview is to give a short summary about our previous and present projects aiming: 1.) to characterize and mitigate toxic side effects of several cytotoxic agents; 2.) to decrease the toxic side effects and improve the antitumor effect of 5-fluorouracil by biochemical modulation and 3.) to study the possibility of individualized drug selection, based on the pharmacobiochemical and pharmacogenetic characteristics of the patients. PMID- 18403294 TI - [European and Hungarian national tasks in oncology]. AB - It is well known that cancer incidence and mortality figures are very poor in Hungary. By providing the latest figures the authors analyze the epidemiological background and the risk factors responsible for this situation. Furthermore, based on international recommendations and national specificities, the authors define areas of action to solve this significant health issue. The main conclusion of their analysis is that it is inevitable to improve oncology care by adjusting it to European standards. The decade-old National Cancer Control Program (NCCP) is improved by incorporating legislative actions, educational issues, research and development priorities. The program now provides the definition of regional centers, recommend improvements of the function of oncology teams and rehabilitation. Based on successful European models, this program must be coordinated by the National Cancer Institute. PMID- 18403295 TI - [The premalignant disease of the endometrium: endometrial intraepithelial neoplasia]. AB - The WHO 1994 classification for endometrial hyperplasias is based on the morphologic features of the lesions. This system characterizes the nuclear cytologic morphology as typical or atypical and describes the glandular architectural pattern as simple or complex. The main problem of this classification is the poor reproducibility. Although the predictive value of the atypical category is high, there are many typical hyperplasia cases with cancer progression. Modern molecular data related to endometrial tumorigenesis and precise computerized morphometric analysis have identified the lesion that may be considered as a precursor of endometrioid adenocarcinoma. By definition, this endometrial intraepithelial neoplasia (EIN) is a clonal proliferation of architecturally and cytologically altered endometrial glands which are prone to malignant transformation to endometrioid (type I) endometrial adenocarcinoma. The morphometric basis of EIN diagnosis is the D-score (DS), which is a logical combination of three morphometric features that represent the glandular complexity, glandular volume and cytological alterations. PTEN inactivation and K ras mutation are the earliest genetic changes that can be revealed in these lesions. Hyperplasia cases that do not fit into the EIN categories are considered as benign or hormonal endometrial hyperplasia. This is the theoretical basis of a new classification system in premalignant endometrial diseases. Retrospective clinical data proved the high predictive value of the EIN scheme, so the decision on therapy can be more established. The reproducibility is excellent with application of precise definitions and PTEN immunohistochemistry. In the "Blue book" published in 2003 the WHO introduces the new morphometric- and molecular based EIN system, and recommends it as an alternative classification method. PMID- 18403296 TI - [Erythropoietic protein supportive treatment in small cell lung cancer]. AB - Anemia is common among patients with malignant tumors, due to the disease and chemotherapy. Anemia decreases patient's quality of life, and worsens the dose intensity of chemotherapy. The aim of this retrospective data-analysis was to determine the rate of transfusions and the maintenance of chemotherapeutic dose intensity in 9 small cell lung cancer patients receiving beta-erythropoietin, due to anemia observed after the first cycle of chemotherapy. The mean pre-treatment hemoglobin concentration of the patients was 116.67+/-8.17 g/L (mean+/-SD). The mean pre-erythropoietin hemoglobin concentration at baseline was 103.11+/-7.52 g/L. Six cycles of platinum compounds and etoposide were used. The post-treatment hemoglobin concentration of patients was 110.11+/-5.37 g/L (p = 0,028 vs. baseline). During these 54 chemotherapeutic cycles, only 2 patients needed transfusion, each of them once. According to our experience, the use of beta erythropoietin in 9 anemic small cell lung cancer patients resulted in a low rate of transfusions and maintenance of cytotoxic treatment dose intensity. The adequate use of beta-erythropoietin is of great help to the physician in the management of small cell lung cancer patients. PMID- 18403297 TI - [Changes in renal function during bisphosphonate treatment of breast cancer patients]. AB - Renal function aberrations during bisphosphonate treatment is a well-known phenomenon. In our retrospective study we examined renal functions of 97 breast cancer patients with bone metastasis during their first year of bisphosphonate treatment i.e. (1) frequency of initial renal function alterations; (2) frequency of decreasing renal function during bisphosphonate treatment; (3) the connection between the laboratory findings and the renal function parameters measured at the beginning of bisphosphonate treatment. At the beginning of bisphosphonate treatment we found a surprisingly high rate (26.80%) of decreased creatinine clearance calculated by the Cockcroft-Gault formula. Decreased creatinine clearance at least once during bisphosphonate treatment has been found in 32.99% of the patients, and in 13.4% of the patients with decreased renal function parameters before bisphosphonates it remained decreased during the one-year period. Expected normal renal function is prognosticated by the renal function parameters and serum calcium level measured before starting bisphosphonate treatment. However, we could not demonstrate any connection between decreasing renal function and either the route of administration or the generation or type of bisphosphonates or the previous use of platinum compounds. Our analysis confirms the necessity of monitoring renal function before and during bisphosphonate treatment, and it is advisable to calculate the creatinine clearance in the upper quarter of the normal range of creatinine levels. In case of decreased renal function, change to a less nephrotoxic bisphosphonate or discontinuing the treatment is suggested. While our results are at variance with the published literature, the above-mentioned questions should be examined in a prospective trial. PMID- 18403298 TI - [Long-term follow-up of patients treated with radical surgery for rectal cancer]. AB - In developed societies colorectal cancer (CRC) is the second most frequent malignant tumor which causes more than 5000 deaths yearly in Hungary. We have attempted to answer the question how to improve the above mentioned data by the long-term follow-up of patients operated upon for rectal cancer at our department. Of the patients operated on for rectal cancer at our department between March 1990 and April 2006, we have conducted regular follow-up of 297 patients according to a protocol developed by us. We have examined the length of time between the rectum operation and the diagnosis and the number of local recurrences, distant metastases, tumor progression in more than one organ as well as second tumors (independent of the rectal cancer). During this period we found 24 local recurrences, 32 distant metastases, 43 tumor progressions in more than one organ, and 21 second tumors. In two patients, in addition to distant metastases, we found a second CRC independent of the original rectal cancer, and in one patient with tumor progression in more than one organ we also detected breast cancer. In one patient we found 3 second tumors (CR, lung and urinary bladder) independent of the original rectal cancer. Altogether we found tumors in 117 out of 297 patients. During the same period, we performed 69/117 operations and 31/117 patients were alive at the end of our study with a median survival of 60.4 (3-184) months. In summary, we can state that this work is beneficial for curing the recurrence of rectal cancer, making the patients' life longer or making the quality of life better for the patients operated on for rectal cancer. PMID- 18403299 TI - [Study of career motivators affecting the emotional labour of health care professionals in oncology]. AB - In the course of their everyday work health care professionals (HCPs) often have to change their true feelings. The literature labels this performance as emotional labor. This article is presenting data on the characteristics of HCPs' most endangered by the negative consequences of emotional labor. Our simple choice question survey was conducted at Debrecen University Medical Healthcare Center with the help of 50 oncology HCPs volunteers. Nearly 90% of the HCPs examined change their true feelings in the course of work. It is very difficult to classify those threatened by the negative upshot of this emotional labor. Due to our research we found appalling differences of work motivation that were tightly interconnected with the respondents' emotional labor and their perceived role/emotional expectations. We succeeded in establishing three clusters and defining each cluster's characteristics. Figures suggest that only somewhat more than the half of the HCPs is authentic professional helper, and 45% of them does not or only slightly perceive the patients' demands concerning their work. Therefore, it is important that the work environment does not only assist the work of HCPs by professional means, but along emotional dimensions as well. PMID- 18403300 TI - [Lipoma of the Fallopian tube]. AB - A swollen fallopian tube was found in a 34 years old woman operated for acute abdominal pain. During this operation the fallopian tube together with the appendix were removed. The histological examination identified appendicitis and a lipoma of the fallopian tube. PMID- 18403301 TI - [The significance of extravasation in oncological care]. AB - The treatment of cancer may be associated with various chemotherapy-induced mucocutaneous reactions. One of the mucocutaneous adverse effects of antineoplastic drugs is the toxic local tissue reaction, the extravasation, which occurs in less than 1-2% of cytotoxic infusions. The standard management of vesicant extravasation includes: discontinuing all local infusions, aspiration of any residual drug, elevating the involved limb, local cooling or warm compresses, local anesthesia, antidotes (sodium thiosulfate for alkylating agents, dimethylsulfoxide (DMSO) for anthracyclines and mitomycin, and hyaluronidase for the vinca alkaloids), and finally surgical debridement with plastic surgery reconstruction. Because the anthracyclines are topoisomerase II poisons that are antagonized by topoisomerase II catalytic inhibitors such as dexrazoxane, it seems to be the treatment of choice immediately after extravasation of doxorubicin, epirubicin, daunorubicin, etc. One systemic dose of dexrazoxane after the accident may significantly reduce the toxic tissue lesions. Repeated intralesional injections of GM-CSF may accelerate the wound healing without the need of skin grafts. PMID- 18403302 TI - [Osteonecrosis of the jaw developing during bisphosphonate treatment]. AB - The article discusses osteonecrosis of the jaw as a possible side effect of bisphosphonate treatment. It provides practical guidelines for prevention, diagnosis and management of bisphosphonate-associated osteonecrosis according to literature and clinical evidence. Since controlled clinical trials have not been carried out, the recommendations are based on reviews, reports and clinical experience. Osteonecrosis of the jaw (ONJ) is a historical clinical entity, which can potentially develop in cancer patients receiving bisphosphonate therapy. The pathogenesis of ONJ has not been totally revealed yet. A thorough dental/oral surgical examination and counseling is recommended in cases when intravenous bisphosphonate therapy is needed. All required dental and surgical treatment should be carried out before starting bisphosphonate therapy to prevent ONJ. The patient should be informed about the possible side effects, and the importance of good oral home care and regular dental check-ups. Once the intravenous bisphosphonate therapy has started, only conservative manipulations should be carried out in the oral cavity. Even in case of developed ONJ, suspension of bisphosphonate therapy is not necessary. In these cases a non-surgical approach is recommended concerning the treatment of ONJ. Regarding the growing number of ONJ cases in association with bisphosphonate therapy it is important for the professionals treating cancer patients to be aware of this phenomenon and the importance of prevention. PMID- 18403303 TI - [Report of the President, Society of Hungarian Oncologists, leaving office (2005 2007)]. PMID- 18403304 TI - [Report of the General Secretary, Society of Hungarian Oncologists, November 2005 to November 2007)]. PMID- 18403305 TI - [Report of the Treasurer, Society of Hungarian Oncologists, November 2005 to November 2007)]. PMID- 18403306 TI - [2007 annual report of the Editor-in-Chief of Magyar Onkologia]. PMID- 18403307 TI - [The necessity, aim and tasks of establishing the Oncology Nursing Section of the Hungarian Cancer Society]. AB - The Hungarian Cancer Society organized its 27th Congress in Budapest on the occasion of its 50th anniversary. The 27th jubilee celebration congress took part in the spirit of this in the Budapest Congress and World Trade Center between November 8-10. The Oncology Nursing part had its turn on 10 November where within the frames of Oncology Nursing the invited could listen to presentations in 3 different sections: Prevention, Active Clinical Oncology Nursing and Cure of Souls-Rehabilitation. I could also call November 10th a day of historic importance since the Nursing Section had the opportunity for reorganization on its 50th anniversary and could hold its statutory meeting as a member of the Society of the same rank. Dr. Jan Foubert Ph.D., the former president and present Executive Director of EONS (European Oncology Nursing Society), honored the congress with his presence, assisting our work with his valuable pieces of advice. As a result of this on December 1, 2007 our Section could join EONS, which is centered in Brussels and has 29 countries as members well-prepared. With the Section having been reorganized, a wide cooperation amongst doctors, oncologists, oncology nurses and further experts could start, where they can all work together now against cancer, a serious problem of our age all over the world. PMID- 18403313 TI - Relationship between insulin resistance and some coagulation and fibrinolytic parameters in patients with metabolic syndrome. AB - BACKGROUND: Insulin resistance syndrome has been shown to be associated with many coagulation and fibrinolytic proteins, and these associations suggest that some coagulation and fibrinolytic proteins have a role in atherothrombotic disorders. This study was conducted to determine the levels of some of the hemostatic parameters in subjects having metabolic syndrome and to correlate these values with the anthropometric and metabolic variables associated with this syndrome. SUBJECTS AND METHODS: The participants were 46 obese nondiabetic subjects, 28 (group 1) who fulfilled the Adult Treatment Panel III criteria of the metabolic syndrome and 18 (group 2) who did not have metabolic syndrome, and a control group of 14 lean subjects (group 3) of matched age and sex. Clinical and laboratory evaluation of the study groups focused on anthropometric measurements (weight, height, body mass index, waist circumference, and sagittal abdominal diameter), blood pressure, and laboratory measurements of fasting plasma glucose, fasting insulin, serum lipids, tissue plasminogen activator (t-PA), antithrombin III activity (AT III), protein C, and von Willebrand factor (vWf) antigen. RESULTS: We observed significant increase in the concentrations of t-PA and vWf antigens in subjects having metabolic syndrome (group 1) compared to the other groups, along with nonsignificant changes in the levels of protein C antigen and AT III activity. Both t-PA and vWf showed significant correlation with HOMA-IR (Homeostatic Model Assessment for Insulin Resistance) as a measure of insulin sensitivity. t-PA concentrations showed significant correlations with most of the variables associated with metabolic syndrome, including waist circumference, body mass index, systolic blood pressure, fasting plasma glucose, fasting insulin, and high-density lipoprotein cholesterol. On the other hand, vWf showed significant correlations with fasting plasma glucose, fasting insulin, and sagittal abdominal diameter, with nonsignificant correlations with the other variables. CONCLUSION: Hemostatic and fibrinolytic parameters should be included in characterization of the insulin resistance syndrome. t-PA and vWf-antigen concentrations were increased in subjects with metabolic syndrome and correlated with the HOMA-IR measure of insulin sensitivity. Because both t-PA and vWf are mainly released from vascular endothelium, these findings could be an indicator of endothelial dysfunction in subjects with insulin resistance and metabolic syndrome. PMID- 18403314 TI - Acute lymphoblastic leukemia with hypereosinophilia and 9p21 deletion: case report and review of the literature. AB - Acute lymphoblastic leukemia (ALL) associated with eosinophilia is very rare, with approximately 44 reported cases. We are reporting this case not only because of the rarity of ALL with peripheral blood eosinophilia, but also because we observed a homozygous deletion of the 9p21 locus corresponding to the p16 gene, a cytogenetic abnormality that was not reported in other documented cases. ALL with eosinophilia must be considered a distinct clini-copathologic entity. It is very important for clinicians to be aware of this specific manifestation of ALL within the context of a persistent peripheral eosinophilia, particularly if no lymphoblasts are present in the peripheral blood. PMID- 18403316 TI - Sadness, depression, and avoidance behavior. AB - Research into genetic, psychosocial, and cognitive explanations for depression (biopsychosocial models) provides support for the role of these variables in the etiology of depression. Regularly identified as basic to depression is loss, and the experience of loss has been found to be more influential than genetic factors in the causation of depression. A distinction is drawn between sadness and depression. Sadness is conceptualized as a normal, time-limited response to loss, whereas depression is a disorder because it is recurring and disruptive. Missing from current biopsychosocial explanations for depression is the importance of avoidance behavior in relation to loss. The thesis of this article is that sadness is transformed into depression as a result of avoidance behavior, which blocks access to lost positive reinforcers. Successful treatment requires replacing avoidance behavior with approach behaviors to overcome the consequences of the loss. An equation is presented specifying the factors involved in depression and how they are related. Implications following from the equation fit well with research results. Treatment issues associated with the theory are discussed. PMID- 18403317 TI - Quantifying size distributions of nanolipoprotein particles with single-particle analysis and molecular dynamic simulations. AB - Self-assembly of purified apolipoproteins and phospholipids results in the formation of nanometer-sized lipoprotein complexes, referred to as nanolipoprotein particles (NLPs). These bilayer constructs are fully soluble in aqueous environments and hold great promise as a model system to aid in solubilizing membrane proteins. Size variability in the self-assembly process has been recognized for some time, yet limited studies have been conducted to examine this phenomenon. Understanding the source of this heterogeneity may lead to methods to mitigate heterogeneity or to control NLP size, which may be important for tailoring NLPs for specific membrane proteins. Here, we have used atomic force microscopy, ion mobility spectrometry, and transmission electron microscopy to quantify NLP size distributions on the single-particle scale, specifically focusing on assemblies with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and a recombinant apolipoprotein E variant containing the N-terminal 22 kDa fragment (E422k). Four discrete sizes of E422k/DMPC NLPs were identified by all three techniques, with diameters centered at approximately 14.5, 19, 23.5, and 28 nm. Computer simulations suggest that these sizes are related to the structure and number of E422k lipoproteins surrounding the NLPs and particles with an odd number of lipoproteins are consistent with the double-belt model, in which at least one lipoprotein adopts a hairpin structure. PMID- 18403319 TI - Symposium: Thomas L. Petty Aspen Lung Conference: lung injury and repair. Introduction and perspective. PMID- 18403318 TI - Intracellular cholesterol transporter StarD4 binds free cholesterol and increases cholesteryl ester formation. AB - StarD4 protein is a member of the StarD4 subfamily of steroidogenic acute regulatory-related lipid transfer (START) domain proteins that includes StarD5 and StarD6, proteins whose functions remain poorly defined. The objective of this study was to isolate and characterize StarD4's sterol binding and to determine in a hepatocyte culture model its sterol transport capabilities. Utilizing purified full-length StarD4, in vitro binding assays demonstrated a concentration dependent binding of [(14)C]cholesterol by StarD4 similar to that of the cholesterol binding START domain proteins StarD1 and StarD5. Other tested sterols showed no detectable binding to StarD4, except for 7alpha-hydroxycholesterol, for which StarD4 demonstrated weak binding on lipid protein overlay assays. Subsequently, an isolated mouse hepatocyte model was used to study the ability of StarD4 to bind/mobilize/distribute cellular cholesterol. Increased expression of StarD4 in primary mouse hepatocytes led to a marked increase in the intracellular cholesteryl ester concentration and in the rates of bile acid synthesis. The ability and specificity of StarD4 to bind cholesterol and, as a function of its level of expression, to direct endogenous cellular cholesterol suggest that StarD4 plays an important role as a directional cholesterol transporter in the maintenance of cellular cholesterol homeostasis. PMID- 18403320 TI - Lung injury and repair. Proceedings of the Thomas L. Petty Aspen Lung Conference 50th Annual Meeting. June 6-9, 2007. Aspen, Colorado, USA. PMID- 18403322 TI - Treatment of acute lung injury: clinical and experimental studies. AB - This presentation at the 2007 Aspen Lung Injury and Repair Conference provided a brief historical perspective from the 1998 Aspen Conference on Acute Lung Injury, highlighting the discussion of clinical definitions. There was also a review of the National Heart, Lung, and Blood Institute ARDS Network clinical trials, with an emphasis on the success of the lung-protective ventilation strategy in reducing mortality. In addition, there was a discussion of the recently completed fluid and catheter treatment trial, which demonstrated no benefit for pulmonary arterial catheterization over central venous catheterization for monitoring patients with acute lung injury (ALI). The trial demonstrated an increase in ventilator-free days with a fluid-conservative protocol. Finally, there was a discussion of recent experimental studies that show promise for cell-based therapy with mesenchymal stem cells for the treatment of endotoxin-induced ALI in mice. There were three objectives for this presentation at the 2007 Lung Injury, Repair, and Remodeling Conference: (1) to provide a brief historical perspective from the 1998 Aspen Conference on Acute Lung Injury; (2) to review major National Heart, Lung, and Blood Institute clinical trials conducted in the past 10 years by the ARDS Network; and (3) to review some of our promising work with cell-based therapy for experimental ALI. PMID- 18403321 TI - Interactions between mechanical and biological processes in acute lung injury. AB - Human studies and animal models suggest that mechanical as well as biological processes contribute to acute lung injury. While mechanical stresses and bacterial products can directly alter the endothelial and epithelial barriers in the lungs, a growing body of evidence suggests that synergistic interactions between low levels of mechanical stress and bacterial products in the lungs can cause or exacerbate acute lung injury. New approaches to disrupting these synergistic interactions between mechanical stress and innate immunity have the potential to reduce the incidence or improve the outcome of acute lung injury in humans. PMID- 18403323 TI - Epithelial progenitor cells of the embryonic lung and the role of microRNAs in their proliferation. AB - The entire epithelium of the lung is generated from a small pool of undifferentiated progenitor cells. At least during the early stages of development these reside in the distal tips of the embryonic lung. They respond to multiple signals from the surrounding mesenchyme and play a critical role as morphogenetic organizing centers. In addition, they proliferate rapidly and give rise to daughter cells that differentiate into all the specialized epithelial cells types of the newborn lung. Despite the importance of the progenitor cells, we still know relatively little about the mechanisms controlling their proliferation, morphogenesis, and developmental fate. Here, we discuss new data on the potential role of microRNAs in co-coordinately regulating multiple signaling pathways in embryonic progenitor cells. In particular, our recent transgenic experiments suggest that microRNAs encoded by the miR-17-92 cluster positively promote proliferation of epithelial progenitor cells and inhibit their differentiation. We speculate on how this information might be exploited therapeutically in the long term. PMID- 18403324 TI - What differentiates normal lung repair and fibrosis? Inflammation, resolution of repair, and fibrosis. AB - There has been ongoing controversy related to what differentiates normal lung repair and fibrosis. For example, the current prevailing concept has been that idiopathic forms of pulmonary fibrosis are due only to epithelial injury in response to some unknown cause that results in persistent evolving fibrosis without preceding inflammation. This concept would suggest that the lung responds to injury in a different manner than other organs, such as the liver, kidney, and heart. However, that would seem to contradict known established pathological concepts. To address this controversy, concepts were presented as follows: (1) loss of basement membrane integrity is critical in determining the "point of no return," and contributes to the inability to reestablish normal lung architecture with promotion of fibrosis; (2) loss of epithelial cells, endothelial cells, and basement membrane integrity in usual interstitial pneumonia associated with idiopathic pulmonary fibrosis leads to destroyed lung architecture and perpetual fibrosis; (3) transforming growth factor-beta is necessary, but not entirely sufficient, to promote permanent fibrosis; (4) persistent injury/antigen/irritant is critical for the propagation of fibrosis; (5) idiopathic pulmonary fibrosis is an example of a process related to the persistence of an "antigen(s)," chronic inflammation, and fibrosis; and (6) unique cells are critical cellular players in the regulation of fibrosis. In keeping with the theme of the Aspen Lung Conference, it is hoped that more questions are raised than answered in this presentation, in support of the continued need for research in this area to address these important concepts. PMID- 18403325 TI - Escape from the matrix: multiple mechanisms for fibroblast activation in pulmonary fibrosis. AB - Lung fibrosis is a recognized feature of many chronic lung diseases and is central to the pathogenesis of idiopathic pulmonary fibrosis, a disease that carries a prognosis worse than many cancers. Current research into this condition is defining the key pathways of activation either in resident fibroblasts, matrix producing cells derived from circulating fibrocytes, or epithelial cells that appear to transdifferentiate to fibroblast-like cells. The downstream signaling pathways are also being delineated as well as the gene interactions leading to altered cell phenotype. These studies have led to an appreciation that multiple pathways, including inflammatory and coagulation cascades, are involved in the pathogenesis of idiopathic pulmonary fibrosis. As these facts come to light, we are exploring promising new approaches to treat fibroses and halt the inexorable progression that is a feature of these disorders. This article reviews these findings and our current concepts of the key molecular events leading to tissue damage and excessive matrix deposition in lung fibrosis. It also highlights the need for new studies to delineate alternative pathogenetic mechanisms and integrate these pathways so we have a framework to better understand their importance in individual patients. PMID- 18403326 TI - Matrix metalloproteinase-induced fibrosis and malignancy in breast and lung. AB - Fibrosis is a pathological condition in which tissue structure is disrupted by production of excess extracellular matrix (ECM), and chronic tissue fibrosis is associated with tumor development. Myofibroblasts are the principal mediators of fibrosis, producing abundant ECM as well as inflammatory and angiogenic factors. Myofibroblasts are also abundant in tumor stroma, where they facilitate tumor growth and progression. Matrix metalloproteinases (MMPs), enzymes that degrade and remodel the ECM, are believed to play a critical role in the development of fibrotic tissue, though the mechanism by which this occurs is unclear. Expression of MMP-3 in mammary epithelial cells of transgenic mice stimulates development of fibrosis and subsequent tumor formation. We have recently determined that exposure of mammary epithelial cells to MMP-3 induces a specialized form of epithelial-mesenchymal transition in which the cells acquire myofibroblast-like characteristics and that this process is dependent upon the generation of cellular reactive oxygen species (ROS). New data from culture models in which MMPs are inducibly expressed in human lung cell lines, and transgenic mouse models in which MMPs are inducibly expressed in lung alveolar epithelial cells, suggest that similar processes likely exist in the lung. PMID- 18403327 TI - Bone marrow-derived cells and stem cells in lung repair. AB - Although it has been many years since publication of the first peer-reviewed studies showing that bone marrow (BM)-derived cells can become mature-appearing epithelial cells, we still know very little regarding the mechanisms, kinetics, cells, and potential clinical utility or pathology associated with this phenomenon. The initial discovery of BM-derived epithelial cells (BMDE) in the liver was published by Petersen and colleagues (Petersen BE, Bowen WC, Patrene KD, Mars WM, Sullivan AK, Murase N, Boggs SS, Greenberger JS, Goff JP. Bone marrow as a potential source of hepatic oval cells. Science 1999;284:1168-1170). Since that time, BMDE were identified in the skin, eye, GI tract, kidney, and the lung. Surprisingly, once several laboratories started to examine the effects of BM cells after tissue injury, BM-derived cells of different types were found to decrease tissue injury and enhance tissue repair, often without engraftment of marrow-derived epithelial cells. Thus, the potentially beneficial effects of BM derived cells in some tissue microenvironments may be unrelated to differentiation into nonhematopoietic cell types. Here, I focus on recent findings from my laboratory as well as several other laboratories on the effects of BM cells on lung damage, and BMDE in the lung, including tracheal epithelial cells, bronchiolar epithelial cells, and type II pneumocytes in the alveoli. Potential mechanisms underlying the appearance of marrow-derived epithelial cells, and the role of tissue damage are discussed. PMID- 18403329 TI - Biology of fibroblasts and myofibroblasts. AB - Despite their importance in fibrosis, the origin of fibroblasts and the genesis of the various subpopulations characterized by distinct phenotypes remain unclear. Various studies have described distinct and relatively stable phenotypes in fibroblasts isolated from lung tissue undergoing remodeling, which were not present in the normal intact tissue. This indicates a process by which these distinct fibroblast subpopulations could arise de novo from resident lung progenitors or precursor cells and/or be recruited from distal organs, such as the bone marrow. Evidence for these possibilities is reviewed, but there is as yet incomplete understanding of the precise precursor cells and the potential interrelationships between the various phenotypes, especially as to how they relate to the distinct myofibroblast phenotype. Moreover, the complexity of the mechanism for the genesis of these phenotypes, such as the myofibroblast, is highlighted by the multilevel regulation of the differentiation process, with evidence for the importance of multiple signaling pathways, transcription factors, and epigenetic mechanisms. Future studies into these various unsettled areas are essential to provide further insights that may help provide the pathway for novel translational approaches. PMID- 18403328 TI - Maintenance and repair of the bronchiolar epithelium. AB - Bronchioles of the distal conducting airway are lined by a simple epithelium composed primarily of nonciliated secretory (Clara) cells and ciliated cells. These cells are long-lived in the normal lung; renewal is mediated by cells that constitute a nonclassical stem cell hierarchy. Within this type of hierarchy, facultative progenitor cells are responsible for normal epithelial maintenance and rare adult tissue-specific stem cells are activated only in response to depletion of the facultative progenitor cell pool. This organizational structure is a departure from the classical stem cell hierarchies that maintain rapidly renewing tissues such as the epithelium of the small intestine. This article compares cellular and molecular mechanisms of epithelial renewal in the relatively quiescent bronchiolar epithelium and in the mitotically active intestinal epithelium. Fundamental distinctions between stem cell hierarchies of slowly and rapidly renewing epithelia are highlighted and may provide insight into tissue-specific interpretation of signals that mediate repair in some tissues but lead to remodeling and chronic disease in other organ systems. PMID- 18403330 TI - Fibrogenesis of parenchymal organs. AB - Fibrosis of parenchymal organs is caused by prolonged injury, deregulation of the normal processes of wound healing, and extensive deposition of extracellular matrix (ECM) proteins. The current review will focus on common features of fibrogenesis in parenchymal organs, and will briefly discuss common features and differences in the pathophysiology of fibrosis. Comparison of hepatic, renal, and pulmonary fibrosis has identified several common mechanisms. Common themes include a critical role for the cytokine transforming growth factor beta and the generation of reactive oxygen species. Activated myofibroblasts are the common cell type that produce the excessive fibrous scar and may originate from endogenous cells such as hepatic stellate cells or fibroblasts, from the bone marrow such as fibrocytes, or from the transition of epithelial cells to mesenchymal cells. These concepts open new prospects for multidisciplinary research and the development of new therapies for fibrosis. PMID- 18403333 TI - Symposium: Thomas L. Petty Aspen Lung Conference: lung injury and repair. Conference summary. PMID- 18403331 TI - Genetic analysis of sporadic and familial interstitial pneumonia. AB - Although much progress has been made in understanding the biology and clinical course of interstitial pneumonia, the etiology of this disease remains elusive. Epidemiologic studies have consistently identified cigarette smoke as an important exposure; however, most smokers do not develop interstitial pneumonia and many individuals with interstitial pneumonia do not smoke cigarettes. Moreover, interstitial pneumonias have been reported to cluster in families. Thus, a more thorough understanding of the genetic etiology of interstitial pneumonia may prove critically important in defining the biology and clinical course of this complex human disease. PMID- 18403332 TI - Making genomics functional: deciphering the genetics of acute lung injury. AB - Acute lung injury (ALI) is a common and frequently devastating illness characterized by acute hypoxemic respiratory failure, profound inflammation, and flooding of the alveoli. Despite recent advances in ALI care, the morbidity and mortality of ALI continues to be unacceptably high. ALI-inciting events (e.g., sepsis, trauma, aspiration, pneumonia) are quite common, yet only a fraction of patients develop the syndrome. This heterogeneity of patients presenting with ALI has sparked interest in identifying the role of genetic factors that contribute to ALI susceptibility and prognosis. Recent advances in high-throughput sequencing and expression technologies now provide the tools to perform large scale genomic analyses in complex disorders such as ALI; gene expression profiling and pathway analysis provide further insight into previously described molecular pathways involved in the syndrome. In this article, we describe the use of genomewide association studies, ortholog in silico techniques, utility of consomic rat methods, and candidate gene approaches using expression profiling and pathway analyses. These methods have confirmed suspected ALI candidate genes (e.g., IL-6 and MIF), but more impressively have identified novel genes (e.g., GADD45alpha and PBEF) not previously suspected in ALI. The analysis of the molecular pathways (e.g., the cytoskeleton in vascular barrier regulation) has identified additional genes contributing to the development and severity of ALI (e.g., MLCK), thereby providing therapeutic targets in this devastating illness. PMID- 18403371 TI - Coupling between ATP binding and DNA cleavage by DNA topoisomerase II: A unifying kinetic and structural mechanism. AB - DNA topoisomerase II is a molecular machine that couples ATP hydrolysis to the transport of one DNA segment through a transient break in another segment. To learn about the energetic connectivity that underlies this coupling, we investigated how the ATPase domains exert control over DNA cleavage. We dissected the DNA cleavage reaction by measuring rate and equilibrium constants for the individual reaction steps utilizing defined DNA duplexes in the presence and absence of the nonhydrolyzable ATP analog 5'-adenylyl-beta,gamma-imidodiphosphate (AMPPNP). Our results revealed the existence of two enzyme conformations whose relative abundance is sensitive to the presence of nucleotides. The predominant species in the absence of nucleotides binds DNA at a diffusion limited rate but cannot efficiently cleave DNA. In the presence of AMPPNP, most of the enzyme is converted to a state in which DNA binding and release is extremely slow but which allows DNA cleavage. A minimal kinetic and thermodynamic framework is established that accounts for the cooperativity of cleavage of the two DNA strands in the presence and absence of bound AMPPNP and includes conformational steps revealed in the kinetic studies. The model unifies available kinetic, thermodynamic, and structural data to provide a description for the reaction in terms of the order and rate of individual reaction steps and the physical nature of the species on the reaction path. Furthermore, this reaction framework provides a foundation for a future in-depth analysis of energy transduction by topoisomerase II, for guiding and interpreting future structural studies, and for analyzing the mechanism of drugs that convert topoisomerase into a cellular poison. PMID- 18403372 TI - Growth factor-induced phosphorylation of sterol regulatory element-binding proteins inhibits sumoylation, thereby stimulating the expression of their target genes, low density lipoprotein uptake, and lipid synthesis. AB - The destiny and activity of sterol regulatory element-binding proteins (SREBPs) in the nucleus are regulated by modification with ubiquitin, small ubiquitin-like modifier (SUMO), or phosphorus. ERK-dependent phosphorylation causes an increase in their transcriptional activity, whereas SUMO modification halts it. We hypothesized a causal linkage between phosphorylation and sumoylation because their sites are very closely located in SREBP-1 and -2 molecules. When Ser(455), a phosphorylation site in SREBP-2, was substituted with Ala, this SREBP-2 mutant was more efficiently modified by SUMO-1. On the other hand, substitution of Asp inhibited SUMO conjugation, mimicking phosphoserine. When cells were cultured with insulin-like growth factor-1, sumoylation of SREBP-2 was decreased with an increase in its phosphorylation, but SREBP-2(S455A) was continuously sumoylated. An ERK cascade inhibitor, U0126, inversely augmented SUMO modification of SREBP 2. Insulin-like growth factor-1 treatment stimulated the expression of SREBP target genes such as the low density lipoprotein (LDL) receptor, squalene synthase, and hydroxymethylglutaryl-CoA synthase genes. These results indicate that growth factor-induced phosphorylation of SREBP-2 inhibits sumoylation, thereby facilitating SREBP transcriptional activity. Glutathione S-transferase pulldown assays revealed that wild-type SREBP-2, but not a mutant lacking Lys(464), interacts with HDAC3 preferentially among the histone deacetylase family members. HDAC3 small interfering RNA induced gene expression of the LDL receptor and thereby augmented fluorescently labeled LDL uptake in HepG2 cells. In summary, growth factors inhibit sumoylation of SREBPs through their phosphorylation, thus avoiding the recruitment of an HDAC3 corepressor complex and stimulating the lipid uptake and synthesis required for cell growth. PMID- 18403373 TI - Involvement of the H3O+-Lys-164 -Gln-161-Glu-345 charge transfer pathway in proton transport of gastric H+,K+-ATPase. AB - Gastric H(+),K(+)-ATPase is shown to transport 2 mol of H(+)/mol of ATP hydrolysis in isolated hog gastric vesicles. We studied whether the H(+) transport mechanism is due to charge transfer and/or transfer of hydronium ion (H(3)O(+)). From transport of [(18)O]H(2)O, 1.8 mol of water molecule/mol of ATP hydrolysis was found to be transported. We performed a molecular dynamics simulation of the three-dimensional structure model of the H(+),K(+)-ATPase alpha subunit at E(1) conformation. It predicts the presence of a charge transfer pathway from hydronium ion in cytosolic medium to Glu-345 in cation binding site 2 (H(3)O(+)-Lys-164 -Gln-161-Glu-345). No charge transport pathway was formed in mutant Q161L, E345L, and E345D. Alternative pathways (H(3)O(+)-Gln-161-Glu-345) in mutant K164L and (H(3)O(+)-Arg-105-Gln-161-Gln-345) in mutant E345Q were formed. The H(+),K(+)-ATPase activity in these mutants reflected the presence and absence of charge transfer pathways. We also found charge transfer from sites 2 to 1 via a water wire and a charge transfer pathway (H(3)O(+)-Asn-794 -Glu-797). These results suggest that protons are charge-transferred from the cytosolic side to H(2)O in sites 2 and 1, the H(2)O comes from cytosolic medium, and H(3)O(+) in the sites are transported into lumen during the conformational transition from E(1)PtoE(2)P. PMID- 18403374 TI - The TpsB translocator HMW1B of haemophilus influenzae forms a large conductance channel. AB - The Haemophilus influenzae HMW1 adhesin is secreted via the two-partner secretion pathway and requires HMW1B for translocation across the outer membrane. HMW1B belongs to the Omp85-TpsB superfamily of transporters and consists of two structural domains, a C-terminal transmembrane beta-barrel and an N-terminal periplasmic domain. We investigated the electrophysiological properties of the purified full-length HMW1B and the C-terminal domain using planar lipid bilayers. Both the full-length and the truncated proteins formed conductive pores with a low open probability, two well defined conductance states, and other substates. The kinetic patterns of the two conductance states were distinct, with rapid and frequent transitions to the small conductance state and occasional and more prolonged openings to the large conductance state. The channel formed by the full length HMW1B showed selectivity for cations, which decreased when measured at pH 5.2, suggesting the presence of acidic residues in the pore. The C-terminal domain of HMW1B was less stable and required reconstitution into liposomes prior to insertion in the bilayer. It formed a channel of smaller conductance but a similar gating pattern as the full-length protein, demonstrating the ability of the last 312 C-terminal amino acids to form a pore and suggesting that the periplasmic domain is not involved in occluding the pore, nor in controlling the inherent basal kinetics of the channel. The HMW1 pro-piece containing the secretion domain, although binding to the channel with high affinity, did not induce channel opening. PMID- 18403375 TI - The impact of supported employment and working on clinical and social functioning: results of an international study of individual placement and support. AB - BACKGROUND: Concerns are frequently expressed that working might worsen the mental health of people with severe mental illness (SMI). Several studies of Individual Placement and Support (IPS), however, have found associations between working and better nonvocational outcomes. IPS has been found to double the return to work of people with SMI in 6 European countries. AIMS: To explore separately associations between IPS, returning to work, and clinical and social outcomes. METHODS: Patients (n = 312) in a randomized controlled trial of IPS in 6 European centers were followed up for 18 months. RESULTS: There were no differences in clinical and social functioning between IPS and control patients at 18 months. Those who worked had better global functioning, fewer symptoms, and less social disability at final follow-up; greater job tenure was associated with better functioning. Working was associated with concurrently better clinical and social functioning, but this contrast was stronger in the control group, suggesting that IPS was better than the control service at helping more unwell patients into work. Working was associated with having been in remission and out of hospital for the previous 6 months. It was also associated with a slight decrease in depression and with being in remission over the subsequent 6 months. CONCLUSIONS: Concerns among clinicians about possible detrimental effects of working and supported employment have been misplaced. Although some of the associations found may have been selection effects, there is sufficient evidence of work having beneficial effects on clinical and social functioning to merit further exploration. PMID- 18403376 TI - Paradoxical air embolism successfully treated with hyperbaric oxygen. AB - The use of the central venous catheter may be complicated by air embolism when central venous pressure is subatmospheric and the catheter is open to the surrounding air. Paradoxical air embolus occurs when the gas bubbles are able to traverse a right to left shunt, gaining access to the systemic arterial circulation causing ischemic symptoms in end organs. In this article, a case of a patient with an unknown patent foramen ovale through which air entered the arterial circulation resulting in obtundation and stroke after inadvertent manipulation of a Hickman catheter is presented. The physiology, clinical manifestations, and management strategies are also discussed. PMID- 18403377 TI - Statistical significance versus clinical significance. AB - Unfortunately, the data were statistically nonsignificant due to insufficient power and poor experimental design. Conversely, many large clinical trials obtain a high level of statistical significance with miniscule differences between groups, which are completely clinically irrelevant. However, with proper marketing, billions can be made from results of dubious clinical importance. In this article, the differences between statistical and clinical significance are briefly discussed. PMID- 18403378 TI - Decision-making algorithm for the STARR procedure in obstructed defecation syndrome: position statement of the group of STARR Pioneers. AB - Internal rectal prolapse (rectal intussusception) and rectocele are frequent clinical findings in patients suffering from refractory constipation that may be best characterized as obstructive defecation syndrome. However, there is still no clear evidence whether the stapled transanal rectal resection (STARR) procedure provides a safe and effective surgical option for symptom resolution in patients with obstructive defecation syndrome, as evidence-based guidelines and functional long-term results are still missing. On the basis of the need for objective evaluation, a European group of experts was founded (Stapled Transanal Rectal Resection Pioneers). Derived from 2 meetings (October 26-28, 2006, Gouvieux, France and November 28-29, 2007, St Gallen, Switzerland) a concept for treatment options in patients suffering from obstructive defecation syndrome was developed, including a clear decision-making algorithm specifically focusing on the role of the stapled transanal rectal resection procedure based on clinical symptoms and dynamic imaging and inclusion and exclusion criteria for the stapled transanal rectal resection procedure. PMID- 18403379 TI - Increased non-stationarity of heart rate during general anaesthesia with sevoflurane or desflurane in children. AB - BACKGROUND: During general anaesthesia, the most prominent change in heart rate variability (HRV) is a decrease in the magnitude of heart rate (HR) oscillation in the high- and low-frequency ranges. In children receiving sevoflurane or desflurane, we observed a significant increase in HR non-stationarity, that is, a significant change of mean HR over time. The aim of our study was to describe this increased non-stationarity and compare it with the decrease in the magnitude of HR oscillation. METHODS: Sixty children received sevoflurane (n=30) or desflurane anaesthesia (n=30). The magnitude of HR oscillation and non stationarity during pre-anaesthesia and anaesthesia were measured by spectral and Hurst analyses using structure function, respectively. RESULTS: Low- and high frequency powers decreased significantly and the very-short-term (218 years and had AF documented on an ECG during the index office visit or during the preceding month. In this survey, 622 ambulatory patients were enrolled (390 males and 232 females; mean age 69.8 +/- 11.8 years). The prevalence of paroxysmal, persistent, and permanent AF was 35, 18, and 47%, respectively. Underlying cardiac disorders present in 513 patients (82%) included hypertensive heart disease (30%), valvular heart disease (27%), coronary artery disease (18%), and myocardial disease (11%). A rate-control strategy was chosen in 53% of the patients (331/622). The mean CHADS(2) score was 1.43 +/- 1.24, and 458/622 patients (73.6%) had a CHADS(2) score >or=1. Among patients with an indication to oral anticoagulant therapy (OAT), 88% (403/458) effectively received it. The rate of OAT was closely correlated with an increasing CHADS(2) score, particularly with patients age (72, 81, and 87% for patients <65, 65-75, and >75 years of age, respectively). True contraindication for OAT was present in 4% (18/458). In the low-risk group (CHADS(2) score = 0), 58% were prescribed OAT, but in 37% of them only for a short period of time (cardioversion/ablation). After a follow-up of 396 +/- 109 days, 72% of the study group (410/570) was still treated by OAT. During follow up, 23/570 patients died (4%), essentially from a cardiovascular cause (15/23), 15 had a non-lethal embolic stroke (2.7%), and 8 had significant bleeding complications (1.5%). CONCLUSION: This study shows one of the highest OAT prescription rates for AF reported until now and demonstrates how successfully guidelines can be applied in the real world. A definite overinterpretation of current guidelines is observed in low-risk patients with AF. True contraindication for OAT (4%) and significant bleeding during OAT (1.5%) were rare. PMID- 18403386 TI - Base over apex: does site matter for pacing the right ventricle? PMID- 18403387 TI - Long-term outcomes in patients with atrioventricular block undergoing septal ventricular lead implantation compared with standard apical pacing. AB - AIMS: Left ventricular function may be altered by right ventricular apical pacing. The aims of the study were to compare the long-term course of different parameters of left ventricular dysfunction in patients undergoing implantation of a dual-chamber pacemaker with the ventricular lead in a septal position vs. in a standard apical position. METHODS: We randomized 98 patients with atrioventricular block (AV-block) undergoing pacemaker implantation to positioning the ventricular lead in the high or mid septum (n = 53) or in the apex (n = 45) of the right ventricle. N-terminal pro-brain natriuetic peptide (BNP) levels, left ventricular ejection fraction (LVEF), and exercise capacity were analysed 3 days, 3 months, and 18 months after the implantation. The primary endpoints were the changes of these parameters from baseline to 18 months. RESULTS: Changes of BNP levels, LVEF, and exercise capacity from baseline to 18 months were statistically not different between septal and apical stimulation. The clinical occurrence or deterioration of overt heart failure was similar in both treatment arms. CONCLUSION: With regard to different parameters of congestive heart failure, a septal stimulation site is not superior to conventional apical pacing in unselected patients undergoing pacemaker implantation for AV-block. PMID- 18403388 TI - Incessant right ventricular outflow tract ventricular tachycardia due to subacute postpartum thyroiditis. PMID- 18403389 TI - Verification of electrical isolation of pulmonary veins following left atrial circumferential ablation may require sinus rhythm. AB - A 67-year-old female with symptomatic paroxysmal atrial fibrillation (AF) underwent left atrial circumferential ablation, and during the procedure, she developed AF. Once the ablation was complete, the left upper pulmonary vein (LUPV) appeared to continue in a rapid disorganized rhythm, despite further attempts at isolating this vein. When the patient was electrically cardioverted to sinus rhythm to assist mapping, the LUPV remained in a disorganized rhythm, pulmonary vein (PV) fibrillation. This case illustrates a possible pitfall in confirming complete isolation of the PVs during AF. PMID- 18403390 TI - Normal thrombin generation in neonates in spite of prolonged conventional coagulation tests. AB - Conventional coagulation tests might be inadequate to explore mechanisms regulating thrombin generation in neonates, because they do not allow full activation of the reduced levels of protein C. Therefore, they do not reflect the action of pro- and anti-coagulants as does the endogenous thrombin potential assessed in the presence of thrombomodulin. Endogenous thrombin potential measured without thrombomodulin was greater than the lower-limit of the adult reference interval in 30% of 109 full-term and 49% of 55 pre-term neonates, a finding consistent with the reduced levels of procoagulants in this setting. When the test was modified adding thrombomodulin, endogenous thrombin potential reverted into the adult reference interval in 97% and 100% full-term and pre-term neonates. In conclusion, the coagulation balance in neonates is restored by the concomitant reduction of pro- and anticoagulants. The restored balance can be shown in vitro by the endogenous thrombin potential test that includes thrombomodulin, but not by conventional coagulation tests. PMID- 18403391 TI - Haplotypes of the EPCR gene, prothrombin levels, and the risk of venous thrombosis in carriers of the prothrombin G20210A mutation. AB - BACKGROUND: Haplotypes A1 and A3 in the endothelial protein C receptor (EPCR) gene are tagged by 4678G/C and 4600A/G respectively. We assessed whether these haplotypes modify the risk of venous thromboembolism in carriers of the prothrombin 20210A allele. DESIGN AND METHODS: We genotyped 4678G/C and 4600A/G in 246 20210A carriers: 84 venous thromboembolism propositi and 162 relatives (13 symptomatic), and in 140 relatives not carrying the 20210A variant. Prothrombin and soluble EPCR (sEPCR) levels were also measured. RESULTS: Among propositi, the mean age at first onset was lower in carriers (35 +/- 8 years) than non-carriers of the 4600G allele (44 +/- 14 years) (p = 0.004). The probability of being free of thrombosis at age 40 was lower in 20210A carriers with the EPCR 4600G allele (p = 0.015). The frequency of the 4600G allele (p=0.002) and the levels of prothrombin antigen (p = 0.002) and sEPCR (p < 0.001) were higher in the propositi than in their asymptomatic relatives. Multivariate analyses showed that the presence of the 4600G allele (OR = 2.5, 95% confidence interval 1.3-5.0), sEPCR > 147 ng/mL (2.8, 1.5-5.2) and prothrombin > 129% (3.8, 1.8-8.3) all increased the thrombotic risk. In bivariate analysis, including the 4600G allele and sEPCR > 147 ng/mL, only the latter remained associated with risk. CONCLUSIONS: These results show that in 20210A carriers the venous thromboembolism risk is influenced both by the actual prothrombin levels and by the EPCR A3 haplotype, via its effect on sEPCR levels. PMID- 18403392 TI - Granulocyte colony-stimulating factor mobilized CFU-F can be found in the peripheral blood but have limited expansion potential. AB - Bone marrow mesenchymal stem cells are multipotent cells found lining the bone marrow cavity supporting the growth and differentiation of hematologic progenitors. There is growing evidence that these cells can, under the right circumstances, enter the peripheral circulation. We show that granulocyte colony stimulating factor mobilized peripheral blood contains cells which form colonies and have a similar fibroblastic morphology (termed CFU-F) to bone marrow mesenchymal stem cells. These cells were found at a very low incidence (0.0002%). Mobilized peripheral blood CFU-F were successfully differentiated into osteogenic and adipogenic lineages. FACS analysis showed that the cells had a similar profile to bone marrow mesenchymal stem cells. Importantly, mobilized peripheral blood CFU-F had limited expansion potential and became senescent 20-25 days after isolation. Mobilized peripheral blood CFU-F also did not have any telomerase activity and displayed significant telomere shortening. The rarity of CFU-F in mobilized peripheral blood and the subsequent pressure to divide in cell culture probably contribute to early cellular senescence. Their potential for use in transplant or gene therapy is, therefore, limited. PMID- 18403393 TI - Severe and moderate hemophilia A: identification of 38 new genetic alterations. AB - Hemophilia A is an X-linked recessive disorder caused by a lack or decrease of factor VIII activity. Its socio-economic impact is high given its high bleeding expression and treatment cost. Our aim was to establish the mutation of each patient to improve family management. A total of 116 unrelated families with severe and moderate hemophilia A were involved. Non-carriers of intron 22 and intron 1 rearrangements were included in F8 gene screening. Intron 1 and 22 inversion frequencies were 3% and 52.5% respectively. Putative mutations were identified in all the families; 38 were new. The cumulative inhibitor incidence was 22%. Approximately half the families carry non-recurrent mutations, which were unique in around one third. Harmful effects for mutations predicting null alleles are expected. Missense mutation consequences are not easily predictable, despite the help of some bio-informatics tools. PMID- 18403394 TI - Phenotype and genotype report on homozygous and heterozygous patients with congenital factor X deficiency. AB - Factor X deficiency is a severe rare hemorrhagic condition inherited as an autosomal recessive trait. It is one of the most severe recessive inherited coagulation disorders. We analyzed the clinical manifestations, laboratory phenotype and genotype in 10 patients with severe Factor X deficiency and in their heterozygous relatives. The most frequent bleeding episodes were hematomas (70%) and gum bleeding (60%). Fifty percent of the homozygous patients required blood transfusion and one-third of heterozygotes required treatment after surgery or delivery. The genetic characterization revealed six different missense mutations, two of which were novel: p.Glu69Lys and p.Asp103His. Haplotype analysis, performed with intra- and extra- FX gene polymorphic markers in Indian, Iranian and Italian patients with the same mutations failed to establish identity by descent, despite the same Caucasian origin. In conclusion, factor X deficiency was confirmed to be one of the most serious among rare bleeding disorders and genetically heterogeneous in different populations. PMID- 18403395 TI - Lower dose rituximab is active in adults patients with idiopathic thrombocytopenic purpura. AB - Rituximab 375 mg/m(2) weekly for four weeks has significant activity in patients with immune thrombocytopenia. We evaluated the activity of lower dose rituximab (100 mg iv weekly for 4 weeks) in 28 adults with idiopathic thrombocytopenic purpura. Overall (platelet count > 50 x 10(9)/L) and complete responses (platelet count > 100 x 10(9)/L) were achieved in 21/28 (75%) and 12/28 (43%) patients respectively. The median time to response and time to complete response were 31 and 44 days respectively. After a median follow-up of 11 months (range 3-18), 7/21 (33%) patients relapsed and 3 needed further treatments. In patients with idiopathic thrombocytopenic purpura, lower dose rituximab seems to show similar activity to standard dose. PMID- 18403396 TI - Resting-state functional connectivity reflects structural connectivity in the default mode network. AB - Resting-state functional connectivity magnetic resonance imaging (fcMRI) studies constitute a growing proportion of functional brain imaging publications. This approach detects temporal correlations in spontaneous blood oxygen level dependent (BOLD) signal oscillations while subjects rest quietly in the scanner. Although distinct resting-state networks related to vision, language, executive processing, and other sensory and cognitive domains have been identified, considerable skepticism remains as to whether resting-state functional connectivity maps reflect neural connectivity or simply track BOLD signal correlations driven by nonneural artifact. Here we combine diffusion tensor imaging (DTI) tractography with resting-state fcMRI to test the hypothesis that resting-state functional connectivity reflects structural connectivity. These 2 modalities were used to investigate connectivity within the default mode network, a set of brain regions--including medial prefrontal cortex (MPFC), medial temporal lobes (MTLs), and posterior cingulate cortex (PCC)/retropslenial cortex (RSC)--implicated in episodic memory processing. Using seed regions from the functional connectivity maps, the DTI analysis revealed robust structural connections between the MTLs and the retrosplenial cortex whereas tracts from the MPFC contacted the PCC (just rostral to the RSC). The results demonstrate that resting-state functional connectivity reflects structural connectivity and that combining modalities can enrich our understanding of these canonical brain networks. PMID- 18403397 TI - Interaction of working memory and long-term memory in the medial temporal lobe. AB - Recent findings indicate that regions in the medial temporal lobe (MTL) do not only play a crucial role in long-term memory (LTM) encoding, but contribute to working memory (WM) as well. However, very few studies investigated the interaction between these processes so far. In a new functional magnetic resonance imaging paradigm comprising both a complex WM task and an LTM recognition task, we found not only that some items were successfully processed in WM but later forgotten, but also that a significant number of items which were not successfully processed in the WM task were subsequently recognized. Activation in the parahippocampal cortex (PHC) during successful WM was predictive of subsequent LTM, but was correlated with subsequent forgetting if the WM task was not successfully solved. The contribution of the PHC to LTM encoding thus crucially depends on whether an item was successfully processed in the WM task. Functional connectivity analysis revealed that across-trial fluctuations in PHC activity were correlated with activation in extensive regions if WM and LTM tasks were correctly solved, whereas connectivity broke down during unsuccessful attempts to do the task, suggesting that activity in the PHC during WM has to be well controlled to support LTM formation. PMID- 18403399 TI - The Dscam homologue of the crustacean Daphnia is diversified by alternative splicing like in insects. AB - In insects, the homologue of the Down syndrome cell adhesion molecule (Dscam) is a unique case of a single-locus gene whose expression has extensive somatic diversification in both the nervous and immune systems. How this situation evolved is best understood through comparative studies. We describe structural, expression, and evolutionary aspects of a Dscam homolog in 2 species of the crustacean Daphnia. The Dscam of Daphnia generates up to 13,000 different transcripts by the alternative splicing of variable exons. This extends the taxonomic range of a highly diversified Dscam beyond the insects. Additionally, we have identified 4 alternative forms of the cytoplasmic tail that generate isoforms with or without inhibitory or activating immunoreceptor tyrosine-based motifs (ITIM and ITAM respectively), something not previously reported in insect's Dscam. In Daphnia, we detected exon usage variability in both the brain and hemocytes (the effector cells of immunity), suggesting that Dscam plays a role in the nervous and immune systems of crustaceans, as it does in insects. Phylogenetic analysis shows a high degree of amino acid conservation between Daphnia and insects except in the alternative exons, which diverge greatly between these taxa. Our analysis shows that the variable exons diverged before the split of the 2 Daphnia species and is in agreement with the nearest-neighbor model for the evolution of the alternative exons. The genealogy of the Dscam gene family from vertebrates and invertebrates confirmed that the highly diversified form of the gene evolved from a nondiversified form before the split of insects and crustaceans. PMID- 18403398 TI - Hearing loss alters the subcellular distribution of presynaptic GAD and postsynaptic GABAA receptors in the auditory cortex. AB - We have shown previously that auditory experience regulates the maturation of excitatory synapses in the auditory cortex (ACx). In this study, we used electron microscopic immunocytochemistry to determine whether the heightened excitability of the ACx following neonatal sensorineural hearing loss (SNHL) also involves pre or postsynaptic alterations of GABAergic synapses. SNHL was induced in gerbils just prior to the onset of hearing (postnatal day 10). At P17, the gamma aminobutyri acid type A (GABA(A)) receptor's beta2/3-subunit (GABA(A)beta2/3) clusters residing at plasma membranes in layers 2/3 of ACx was reduced significantly in size (P < 0.05) and number (P < 0.005), whereas the overall number of immunoreactive puncta (intracellular + plasmalemmal) remained unchanged. The reduction of GABA(A)beta2/3 was observed along perikaryal plasma membranes of excitatory neurons but not of GABAergic interneurons. This cell specific change can contribute to the enhanced excitability of SNHL ACx. Presynaptically, GABAergic axon terminals were significantly larger but less numerous and contained 47% greater density of glutamic acid decarboxylase immunoreactivity (P < 0.05). This suggests that GABA synthesis may be upregulated by a retrograde signal arising from lowered levels of postsynaptic GABA(A)R. Thus, both, the pre- and postsynaptic sides of inhibitory synapses that form upon pyramidal neurons of the ACx are regulated by neonatal auditory experience. PMID- 18403400 TI - Real life experience confirms sustained response to long-term biologics and switching in ankylosing spondylitis. AB - OBJECTIVE: To investigate the long-term response to biological therapies in AS in a real life clinical setting and to quantify non-response and response to 'switching' therapies in these cases. METHODS: All patients prescribed TNF blocking therapies for AS between 1999 and 2006 were studied. Response was evaluated using Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI) and CRP results. RESULTS: A total of 113 patients (84 males: 29 females, mean age 45 yrs, median disease duration 16 yrs, 87% HLA-B27 positive) were identified. At baseline they had a mean BASDAI of 6.57, BASFI 6.57 and CRP of 31 g/dl. At the end of follow-up, these values had reduced to mean BASDAI of 3.12, BASFI 4.16 and CRP of 7 g/dl. Improvements were sustained for 24 months and beyond with no loss of effect. Only nine patients (8%) suffered side-effects leading to cessation or switching of first-line therapy and non-response occurred in 15 patients (13%) in the long term. Fifteen patients (13%) switched to a second drug and 14 of these (93%) had a significant and sustained response. Outcomes were similar regardless of drug used, duration of disease and HLA-B27 status. CONCLUSION: Treatment of active AS with TNF blockers according to the British Society of Rheumatology guidelines leads to a sustained response for over 2 yrs with most patients tolerating the drugs well. The rate of non-response is significantly lower than that seen in RA and nearly all of these patients respond well to a second-line agent. PMID- 18403401 TI - Myocardial ischaemia in patients with primary APS: a 13N-ammonia PET assessment. AB - OBJECTIVE: Evaluate the presence and severity of myocardial ischaemia in a population of asymptomatic patients with primary APS (PAPS) using (13)N-ammonia PET. METHODS: We studied 36 patients, 18 with a diagnosis of PAPS and 18 healthy volunteers. All patients underwent a two-phase (rest-stress) (13)N-ammonia PET. Myocardial perfusion images were acquired and then analysed by two experts in the field. RESULTS: We found ischaemia in 7/18 asymptomatic PAPS patients (38.8%). The anterolateral wall was the most commonly affected cardiac territory [5/7 PAPS patients (71.4%)]. In a severity analysis, we found that five patients (71.4%) had mild ischaemia, one patient (14.2%) had moderate ischaemia and another one (14.2%) had severe defects. All the healthy volunteers studied showed normal myocardial perfusion images. CONCLUSION: An important proportion of PAPS patients, even when asymptomatic, showed myocardial perfusion defects assessed with PET. Most of the ischaemic patients had mild defects and the anterolateral wall was the territory mainly affected. PMID- 18403402 TI - Single-point acupuncture and physiotherapy for the treatment of painful shoulder: a multicentre randomized controlled trial. AB - OBJECTIVE: Evaluate the efficacy of acupuncture associated with physiotherapy for patients with painful shoulder. METHODS: In a multicentre controlled randomized study, participants were recruited with a clinical diagnosis of unilateral subacromial syndrome from six rehabilitation medicine departments belonging to the Public Health System in two Spanish regions. All participants received 15 sessions of physiotherapy during the 3 weeks that the treatment lasted and were randomized to additionally receive, once a week, acupuncture or mock TENS (transcutaneous electrical nerve stimulation). The primary outcome measure was the change in the Constant-Murley Score (CMS) for functional assessment of the shoulder, at 4 weeks after randomization. This study is registered as an International Standard Randomized Controlled Trial, number ISRCTN28687220. RESULTS: A total of 425 patients were recruited. The mean score (s.d.) on the CMS had increased by 16.6 (15.6) points among the acupuncture group, compared with 10.6 (13.5) points in the control group, and the mean difference between the two groups was statistically significant (6.0 points; 95% CI 3.2, 8.8 points; P < 0.001). By the end of the treatment, 53% of the patients in the acupuncture group had decreased their consumption of analgesics, compared with a corresponding 30% among the control group (P < 0.001). CONCLUSIONS: Single-point acupuncture in association with physiotherapy improves shoulder function and alleviates pain, compared with physiotherapy as the sole treatment. This improvement is accompanied by a reduction in the consumption of analgesic medicaments. PMID- 18403404 TI - Effectiveness of infliximab in the treatment of refractory juvenile dermatomyositis with calcinosis. AB - OBJECTIVES: Some juvenile dermatomyositis (JDM) patients have a disease course which is refractory to multiple drug treatments. Prolonged disease activity is associated with increased mortality and morbidity. TNF-alpha has been identified in high levels in JDM patients who have a long disease course and calcinosis. We assessed the response of five refractory JDM patients to the anti-TNF-alpha monoclonal antibody, infliximab. METHODS: For all five patients intravenous infliximab was initially given at a dose of 3 mg/kg. Further doses were then given at weeks 2, 6 and every 8 weeks thereafter. The dose and frequency were tailored in accordance with clinical response. Clinical and laboratory data were collected prospectively. RESULTS: We report results between 8 and 30 months after starting infliximab. Improvements were seen in all five patients as shown by positive changes in physician visual analogue scale (VAS), Childhood Myositis Assessment Score (CMAS), Childhood Health Assessment Questionnaire (CHAQ), joint range of movement and, in some, regression of calcinosis and skin signs. There were no major side effects observed with addition of infliximab to the therapeutic regime. CONCLUSIONS: Major clinical benefit was demonstrated after the initiation of infliximab in all five cases of refractory JDM. PMID- 18403403 TI - Patient perceptions about illness self-management in ANCA-associated small vessel vasculitis. AB - OBJECTIVES: To characterize patient perceptions, related to eight self-management behaviours relevant for adults with ANCA-associated small vessel vasculitis (ANCA SVV), and to determine if these perceptions were associated with performance of each behaviour. METHODS: Adults with ANCA-SVV (n = 202) completed a self administered questionnaire that assessed eight self-management behaviours (adherence to recommendations for medication, health service use, diet, exercise, infection avoidance and symptom monitoring; prompt reporting of symptoms and side effects; and adjusting activities in response to symptoms), perceptions about these behaviours, socio-demographics, clinical factors and social desirability bias. Descriptive statistics were generated to characterize patients' perceptions about difficulty of, importance of, and specific barriers to performing each behaviour. Regression analyses explored whether these variables were associated with performing each behaviour, controlling for potential confounders. RESULTS: With few exceptions, higher perceived importance and lower perceived difficulty of each behaviour were associated with more frequent performance of the behaviour. For each behaviour, several specific barriers were frequently endorsed by patients and a number of these were associated with lower levels of self management. CONCLUSION: This study reveals that patient perceptions about the illness and its treatment influence ANCA-SVV self-management. Perceived barriers to medication, health services, diet and exercise adherence were similar to those in other illnesses. This study also provides insight into barriers experienced by patients in performing behaviours (infection avoidance, symptom monitoring, reporting symptoms and side-effects and adjusting activities) not often previously studied. How the identification of these barriers can help inform future interventions for ANCA-SVV patients is to be discussed. PMID- 18403405 TI - Diffusive sampling of C7-C16 hydrocarbons in workplace air: uptake rates, wall effects and use in oil mist measurements. AB - The measurement of semi-volatile hydrocarbons in workplace air is complicated by their readiness to condense to form aerosols or adsorb on to surfaces. The diffusive sampling and analysis by thermal desorption of alkanes up to hexadecane was investigated with the aim of quantifying vapour from petroleum distillate fractions and possibly differentiating particles from vapour in oil mist measurements of light mineral oil-based metalworking fluids. Diffusive uptake rates were measured on Perkin Elmer thermal desorption tube samplers packed with Tenax TA, and the potential for deposition within the tubes was examined. Hydrocarbon vapour was found to adsorb on the oxide layer that can develop on the sampler's internal walls. General measurements of mixed hydrocarbon vapours (i.e. petroleum distillate fractions) should not be unduly affected if concentrations are greater than approximately 5 mg m(-3) and the tubes are in good condition. For the purposes of differentiating light mineral oil mist and vapour from a total hydrocarbon measurement, it is unlikely that measuring the vapour separately could be used to calculate mist concentrations <3 mg m(-3) with sufficient accuracy. PMID- 18403406 TI - How much of the data published in observational studies of the association between diet and prostate or bladder cancer is usable for meta-analysis? AB - Epidemiologic investigations often report dose-response associations, which may be combined in meta-analyses. The authors examined how often the log odds, risk, or hazard ratio per unit increase in exposure, and its standard error, can be estimated from results reported from observational studies of diet and prostate or bladder cancer so that results are usable in meta-analyses estimating dose response associations. Eight electronic databases were searched for studies reporting on the association of diet, nutrition, or physical activity with these cancers. A total of 767 papers reported 3,284 results; 1,999 (61%) results, reported in 545 (71%) papers, were usable in dose-response meta-analyses. The most important reason that results were not usable was the absence of sufficient information on exposure levels in the different groups. The proportion of results usable in "high-low" meta-analyses (comparisons of extreme categories) was similar (62%). Results that showed evidence of an association were more likely to be usable than results that found no such evidence. Insufficient detail in reporting of results of observational studies can lead to exclusion of these results from meta-analyses and is an important threat to the validity of systematic reviews of such research. Results providing evidence of associations may be overrepresented in meta-analyses of observational studies. PMID- 18403407 TI - Wollknauel is required for embryo patterning and encodes the Drosophila ALG5 UDP glucose:dolichyl-phosphate glucosyltransferase. AB - N-linked glycosylation is a prevalent protein modification in eukaryotic cells. Although glycosylation plays an important role in cell signaling during development, a role for N-linked glycosylation in embryonic patterning has not previously been described. In a screen for maternal factors involved in embryo patterning, we isolated mutations in Drosophila ALG5, a UDP-glucose:dolichyl phosphate glucosyltransferase. Based on the embryonic cuticle phenotype, we designated the ALG5 locus wollknauel (wol). Mutations in wol result in posterior segmentation phenotypes, reduced Dpp signaling, as well as impaired mesoderm invagination and germband elongation at gastrulation. The segmentation phenotype can be attributed to a post-transcriptional effect on expression of the transcription factor Caudal, whereas wol acts upstream of Dpp signalin by regulating dpp expression. The wol/ALG5 cDNA was able to partially complement the hypoglycosylation phenotype of alg5 mutant S. cerevisiae, whereas the two wol mutant alleles failed to complement. We show that reduced glycosylation in wol mutant embryos triggers endoplasmic reticulum stress and the unfolded protein response (UPR). As a result, phosphorylation of the translation factor eIF2alpha is increased. We propose a model in which translation of a few maternal mRNAs, including caudal, are particularly sensitive to increased eIF2alpha phosphorylation. According to this view, inappropriate UPR activation can cause specific patterning defects during embryo development. PMID- 18403408 TI - Dkk1 and Wnt3 interact to control head morphogenesis in the mouse. AB - Loss of Dkk1 results in ectopic WNT/beta-catenin signalling activity in the anterior germ layer tissues and impairs cell movement in the endoderm of the mouse gastrula. The juxtaposition of the expression domains of Dkk1 and Wnt3 is suggestive of an antagonist-agonist interaction. The downregulation of Dkk1 when Wnt3 activity is reduced reveals a feedback mechanism for regulating WNT signalling. Compound Dkk1;Wnt3 heterozygous mutant embryos display head truncation and trunk malformation, which are not found in either Dkk1(+/-) or Wnt3(+/-) embryos. Reducing the dose of Wnt3 gene in Dkk1(-/-) embryos partially rescues the truncated head phenotype. These findings highlight that head development is sensitive to the level of WNT3 signalling and that DKK1 is the key antagonist that modulates WNT3 activity during anterior morphogenesis. PMID- 18403410 TI - Directional migration of neural crest cells in vivo is regulated by Syndecan 4/Rac1 and non-canonical Wnt signaling/RhoA. AB - Directed cell migration is crucial for development, but most of our current knowledge is derived from in vitro studies. We analyzed how neural crest (NC) cells migrate in the direction of their target during embryonic development. We show that the proteoglycan Syndecan-4 (Syn4) is expressed in the migrating neural crest of Xenopus and zebrafish embryos. Loss-of-function studies using an antisense morpholino against syn4 show that this molecule is required for NC migration, but not for NC induction. Inhibition of Syn4 does not affect the velocity of cell migration, but significantly reduces the directional migration of NC cells. Furthermore, we show that Syn4 and PCP signaling control the directional migration of NC cells by regulating the direction in which the cell protrusions are generated during migration. Finally, we perform FRET analysis of Cdc42, Rac and RhoA in vitro and in vivo after interfering with Syn4 and PCP signaling. This is the first time that FRET analysis of small GTPases has been performed in vivo. Our results show that Syn4 inhibits Rac activity, whereas PCP signaling promotes RhoA activity. In addition, we show that RhoA inhibits Rac in NC cells. We present a model in which Syn4 and PCP control directional NC migration by, at least in part, regulating membrane protrusions through the regulation of small GTPase activities. PMID- 18403409 TI - Wt1 negatively regulates beta-catenin signaling during testis development. AB - beta-Catenin, as an important effector of the canonical Wnt signaling pathway and as a regulator of cell adhesion, has been demonstrated to be involved in multiple developmental processes and tumorigenesis. beta-Catenin expression was found mainly on the Sertoli cell membrane starting from embryonic day 15.5 in the developing testes. However, its potential role in Sertoli cells during testis formation has not been examined. To determine the function of beta-catenin in Sertoli cells during testis formation, we either deleted beta-catenin or expressed a constitutively active form of beta-catenin in Sertoli cells. We found that deletion caused no detectable abnormalities. However, stabilization caused severe phenotypes, including testicular cord disruption, germ cell depletion and inhibition of Mullerian duct regression. beta-Catenin stabilization caused changes in Sertoli cell identity and misregulation of inter-Sertoli cell contacts. As Wt1 conditional knockout in Sertoli cells causes similar phenotypes to our stabilized beta-catenin mutants, we then investigated the relationship of Wt1 and beta-catenin in Sertoli cells and found Wt1 inhibits beta-catenin signaling in these cells during testis development. Wt1 deletion resulted in upregulation of beta-catenin expression in Sertoli cells both in vitro and in vivo. Our study indicates that Sertoli cell expression of beta-catenin is dispensable for testis development. However, the suppression of beta-catenin signaling in these cells is essential for proper testis formation and Wt1 is a negative regulator of beta-catenin signaling during this developmental process. PMID- 18403411 TI - The Arabidopsis OBERON1 and OBERON2 genes encode plant homeodomain finger proteins and are required for apical meristem maintenance. AB - Maintenance of the stem cell population located at the apical meristems is essential for repetitive organ initiation during the development of higher plants. Here, we have characterized the roles of OBERON1 (OBE1) and its paralog OBERON2 (OBE2), which encode plant homeodomain finger proteins, in the maintenance and/or establishment of the meristems in Arabidopsis. Although the obe1 and obe2 single mutants were indistinguishable from wild-type plants, the obe1 obe2 double mutant displayed premature termination of the shoot meristem, suggesting that OBE1 and OBE2 function redundantly. Further analyses revealed that OBE1 and OBE2 allow the plant cells to acquire meristematic activity via the WUSCHEL-CLAVATA pathway, which is required for the maintenance of the stem cell population, and they function parallel to the SHOOT MERISTEMLESS gene, which is required for preventing cell differentiation in the shoot meristem. In addition, obe1 obe2 mutants failed to establish the root apical meristem, lacking both the initial cells and the quiescent center. In situ hybridization revealed that expression of PLETHORA and SCARECROW, which are required for stem cell specification and maintenance in the root meristem, was lost from obe1 obe2 mutant embryos. Taken together, these data suggest that the OBE1 and OBE2 genes are functionally redundant and crucial for the maintenance and/or establishment of both the shoot and root meristems. PMID- 18403412 TI - MALS-3 regulates polarity and early neurogenesis in the developing cerebral cortex. AB - Apicobasal polarity plays an important role in regulating asymmetric cell divisions by neural progenitor cells (NPCs) in invertebrates, but the role of polarity in mammalian NPCs is poorly understood. Here, we characterize the function of the PDZ domain protein MALS-3 in the developing cerebral cortex. We find that MALS-3 is localized to the apical domain of NPCs. Mice lacking all three MALS genes fail to localize the polarity proteins PATJ and PALS1 apically in NPCs, whereas the formation and maintenance of adherens junctions appears normal. In the absence of MALS proteins, early NPCs progressed more slowly through the cell cycle, and their daughter cells were more likely to exit the cell cycle and differentiate into neurons. Interestingly, these effects were transient; NPCs recovered normal cell cycle properties during late neurogenesis. Experiments in which MALS-3 was targeted to the entire membrane resulted in a breakdown of apicobasal polarity, loss of adherens junctions, and a slowing of the cell cycle. Our results suggest that MALS-3 plays a role in maintaining apicobasal polarity and is required for normal neurogenesis in the developing cortex. PMID- 18403414 TI - Maximal oxygen intake and independence in old age. AB - This brief review examines the likelihood that a deterioration of aerobic fitness will lead to a loss of independence in old age. The rate of deterioration of maximal aerobic power observed in middle-aged adults continues unabated during the retirement years. Loss of independence seems likely if maximal oxygen intake falls below a threshold of 18 ml/[kg x min] in men and 15 ml/[kg x min] in women, reached at 80-85 years. A regular programme of aerobic exercise can slow or reverse the functional deterioration, reducing the individual's biological age by 10 or more years, and potentially prolonging independence by a similar amount. There remains a need to clarify the importance of decreasing aerobic fitness relative to other potential causes of dependency but, from the practical viewpoint, regular aerobic activity should be commended to elderly people since it can address many of the issues of both functional loss and chronic disease. PMID- 18403413 TI - Controlling morpholino experiments: don't stop making antisense. AB - One of the most significant problems facing developmental biologists who do not work on an organism with well-developed genetics - and even for some who do - is how to inhibit the action of a gene of interest during development so as to learn about its normal biological function. A widely adopted approach is to use antisense technologies, and especially morpholino antisense oligonucleotides. In this article, we review the use of such reagents and present examples of how they have provided insights into developmental mechanisms. We also discuss how the use of morpholinos can lead to misleading results, including off-target effects, and we suggest controls that will allow researchers to interpret morpholino experiments correctly. PMID- 18403415 TI - Dose-response relationship between physical activity and mental health: the Scottish Health Survey. AB - OBJECTIVES: Regular physical activity is thought to be associated with better mental health, although there is a lack of consensus regarding the optimal amount and type of activity to achieve these benefits. The association between mental health and physical activity behaviours was examined among a representative sample of men and women from the Scottish Health Surveys. METHODS: Self-reported physical activity was measured and the General Health Questionnaire (GHQ-12) was administered in order to obtain information on current mental health. Participants were 19 842 men and women. Risk estimates per category of physical activity sessions per week were calculated using logistic regression models. RESULTS: Psychological distress (based on a score of 4 or more on the GHQ-12) was evident in 3200 participants. Any form of daily physical activity was associated with a lower risk of psychological distress after adjustment for age, gender, social economic group, marital status, body mass index, long-standing illness, smoking and survey year (OR 0.59, 95% CI 0.52 to 0.66, p<0.001). A dose-response relationship was apparent, with moderate reductions in psychological distress with less frequent activity (OR 0.67, 95% CI 0.61 to 0.75). Different types of activities including domestic (housework and gardening), walking and sports were all independently associated with lower odds of psychological distress, although the strongest effects were observed for sports (OR 0.67, 95% CI 0.54 to 0.82). CONCLUSION: Mental health benefits were observed at a minimal level of at least 20 min/week of any physical activity, although a dose-response pattern was demonstrated with greater risk reduction for activity at a higher volume and/or intensity. PMID- 18403416 TI - Does occupational success influence longevity among England test cricketers? AB - OBJECTIVE: To determine whether the longevity of England test cricketers is influenced by occupational success, controlling for the influence of social background. DESIGN: Archival survey. SETTING: England. SUBJECTS: Those 418 cricketers who played for England in a test match from the first test in 1876 to 1963 when the distinction between amateur and professional status was removed. MAIN OUTCOME MEASURES: Length of life. RESULTS: Survival analysis of players born between 1827 and 1941 (349 dead, 69 alive) showed a significant relation between mortality and year of birth (p<0.001), amateur/professional status (p = 0.042) and the number of test matches played (p = 0.042). Captaining England was not related to survival. CONCLUSION: The link between longevity and both social background and occupational success is supported among test match cricketers. Amateur (or "gentlemen") cricketers from more privileged social backgrounds survived longer than professionals (or "players"). The most successful cricketers who played in a larger number of tests lived longer than those who played in a smaller number of tests. Captaining England, which could be regarded as a form of occupational "control", was not associated with longevity. PMID- 18403417 TI - TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages. AB - Transcription in eukaryotes is a multistep process involving the assembly and disassembly of numerous inter- and intramolecular interactions between transcription factors and nucleic acids. The roles of each of these interactions and the regions responsible for them have been identified and studied primarily by the use of mutants, which destroy the inherent properties of the interacting surface. A less intrusive but potentially effective way to study the interactions as well as the surfaces responsible for them is the use of RNA aptamers that bind to the interacting factors. Here, we report the isolation and characterization of high-affinity RNA aptamers that bind to the yeast general transcription factor TFIIB. These aptamers fall into two classes that interfere with TFIIB's interactions with either TBP or RNA polymerase II, both of which are crucial for transcription in yeast. We demonstrate the high affinity and specificity of these reagents, their effect on transcription and preinitiation complex formation and discuss their potential use to address mechanistic questions in vitro as well as in vivo. PMID- 18403418 TI - The three SoxC proteins--Sox4, Sox11 and Sox12--exhibit overlapping expression patterns and molecular properties. AB - The group C of Sry-related high-mobility group (HMG) box (Sox) transcription factors has three members in most vertebrates: Sox4, Sox11 and Sox12. Sox4 and Sox11 have key roles in cardiac, neuronal and other major developmental processes, but their molecular roles in many lineages and the roles of Sox12 remain largely unknown. We show here that the three genes are co-expressed at high levels in neuronal and mesenchymal tissues in the developing mouse, and at variable relative levels in many other tissues. The three proteins have conserved remarkable identity through evolution in the HMG box DNA-binding domain and in the C-terminal 33 residues, and we demonstrate that the latter residues constitute their transactivation domain (TAD). Sox11 activates transcription several times more efficiently than Sox4 and up to one order of magnitude more efficiently than Sox12, owing to a more stable alpha-helical structure of its TAD. This domain and acidic domains interfere with DNA binding, Sox11 being most affected and Sox4 least affected. The proteins are nevertheless capable of competing with one another in reporter gene transactivation. We conclude that the three SoxC proteins have conserved overlapping expression patterns and molecular properties, and might therefore act in concert to fulfill essential roles in vivo. PMID- 18403419 TI - Flexible GnRH antagonist versus flare-up GnRH agonist protocol in poor responders treated by IVF: a randomized controlled trial. AB - BACKGROUND: Although initial studies in poor responders using GnRH antagonists have reported encouraging results, they are limited in number, only a few of them are prospective, while the majority is characterized by limited power to detect a clinically important difference. METHODS: A randomized controlled trial was performed in patients with one or more previous failed IVF cycles in which five or less oocytes were retrieved, using > or =300 IU of gonadotrophins/day. Patients were randomized by computer-generated list and treated by either the flare-up GnRH agonist protocol (n = 90) or a flexible GnRH antagonist protocol (n = 180). RESULTS: Ongoing pregnancy rate, the primary outcome measure, was significantly higher in the antagonist group compared with the agonist group (12.2 versus 4.4%, P< 0.048; difference 7.8%, 95% CI: 0.2 to 14.0). Estradiol levels on the day of hCG administration were lower in the antagonist protocol [median (interquartile range): 572 (325-839) versus 727 (439-1029) pg/ml, P = 0.018]. Clinical and biochemical pregnancy rates, fertilization and implantation rates, as well as the number of oocytes retrieved, the number of mature oocytes present, the stimulation period and the gonadotrophin dosage were not significantly different between the two groups compared. CONCLUSIONS: The flexible GnRH antagonist protocol is associated with significantly higher ongoing pregnancy rates compared with the flare-up GnRH agonist protocol in poor responders. PMID- 18403420 TI - Selective modulation of interactions between ventral premotor cortex and primary motor cortex during precision grasping in humans. AB - In humans, the rostral part of the ventral premotor cortex (PMv), the homologue of F5 in monkeys, is known to be critically involved in shaping the hand to grasp objects. How does information about hand posture, that is processed in PMv, give rise to appropriate motor commands for transmission to spinal circuits controlling the hand? Whereas PMv is crucial for skilled visuomotor control of the hand, PMv sends relatively few direct corticospinal projections to spinal segments innervating hand muscles and the most likely route for PMv to contribute to the control of hand shape is through cortico-cortical connections with primary motor cortex (M1). If this is the case, we predicted that PMv-M1 interactions should be modulated specifically during precision grasping in humans. To address this issue, we investigated PMv-M1 connections by means of paired-pulse transcranial magnetic stimulation (TMS) and compared whether they were differentially modulated at rest, and during precision versus power grip. To do so, TMS was applied over M1 either in isolation or after a conditioning stimulus delivered, at different delays, over the ipsilateral PMv. For the parameters of TMS tested, we found that, at rest, PMv exerted a net inhibitory influence on M1 whereas, during power grip, this inhibition disappeared and was converted into a net facilitation during precision grip. The finding that, in humans, PMv-M1 interactions are selectively modulated during specific types of grasp provides further evidence that these connections play an important role in control of the hand. PMID- 18403421 TI - Interpolated twitches in fatiguing single mouse muscle fibres: implications for the assessment of central fatigue. AB - An electrically evoked twitch during a maximal voluntary contraction (twitch interpolation) is frequently used to assess central fatigue. In this study we used intact single muscle fibres to determine if intramuscular mechanisms could affect the force increase with the twitch interpolation technique. Intact single fibres from flexor digitorum brevis of NMRI mice were dissected and mounted in a chamber equipped with a force transducer. Free myoplasmic [Ca2+] ([Ca2+](i)) was measured with the fluorescent Ca2+ indicator indo-1. Seven fibres were fatigued with repeated 70 Hz tetani until 40% initial force with an interpolated pulse evoked every fifth tetanus. Results showed that the force generated by the interpolated twitch increased throughout fatigue, being 9 +/- 1% of tetanic force at the start and 19 +/- 1% at the end (P < 0.001). This was not due to a larger increase in [Ca2+](i) induced by the interpolated twitch during fatigue but rather to the fact that the force-[Ca2+](i) relationship is sigmoidal and fibres entered a steeper part of the relationship during fatigue. In another set of experiments, we observed that repeated tetani evoked at 150 Hz resulted in more rapid fatigue development than at 70 Hz and there was a decrease in force ('sag') during contractions, which was not observed at 70 Hz. In conclusion, the extent of central fatigue is difficult to assess and it may be overestimated when using the twitch interpolation technique. PMID- 18403422 TI - The effects of experimental muscle and skin pain on the static stretch sensitivity of human muscle spindles in relaxed leg muscles. AB - Animal studies have shown that noxious inputs onto gamma-motoneurons can cause an increase in the activity of muscle spindles, and it has been proposed that this causes a fusimotor-driven increase in muscle stiffness that is believed to underlie many chronic pain syndromes. To test whether experimental pain also acts on the fusimotor system in humans, unitary recordings were made from 19 spindle afferents (12 Ia, 7 II) located in the ankle and toe extensors or peronei muscles of awake human subjects. Muscle pain was induced by bolus intramuscular injection of 0.5 ml 5% hypertonic saline into tibialis anterior (TA); skin pain was induced by 0.2 ml injection into the overlying skin. Changes in fusimotor drive to the muscle spindles were inferred from changes in the mean discharge frequency and discharge variability of spindle endings in relaxed muscle. During muscle pain no afferents increased their discharge activity: seven afferents (5 Ia, 2 II) showed a decrease and six (4 Ia, 2 II) afferents were not affected. During skin pain of 13 afferents discharge rate increased in one (Ia) and decreased in two (1 Ia, 1 II). On average, the overall discharge rate decreased during muscle pain by 6.1% (P < 0.05; Wilcoxon), but remained essentially the same during skin pain. There was no detectable correlation between subjective pain level and the small change in discharge rate of muscle spindles. Irrespective of the type of pain, discharge variability parameters were not influenced (P > 0.05; Wilcoxon). We conclude that, contrary to the 'vicious cycle' hypothesis, acute activation of muscle or skin nociceptors does not cause a reflex increase in fusimotor drive in humans. Rather, our results are more aligned with the pain adaptation model, based on clinical studies predicting pain-induced reductions of agonist muscle activity. PMID- 18403423 TI - Non-selective beta-adrenergic blockade prevents reduction of the cerebral metabolic ratio during exhaustive exercise in humans. AB - Intense exercise decreases the cerebral metabolic ratio of oxygen to carbohydrates [O(2)/(glucose + (1/2)lactate)], but whether this ratio is influenced by adrenergic stimulation is not known. In eight males, incremental cycle ergometry increased arterial lactate to 15.3 +/- 4.2 mm (mean +/- s.d.) and the arterial-jugular venous (a-v) difference from -0.02 +/- 0.03 mm at rest to 1.0 +/- 0.5 mm (P < 0.05). The a-v difference for glucose increased from 0.7 +/- 0.3 to 0.9 +/- 0.1 mm (P < 0.05) at exhaustion and the cerebral metabolic ratio decreased from 5.5 +/- 1.4 to 3.0 +/- 0.3 (P < 0.01). Administration of a non selective beta-adrenergic (beta(1) + beta(2)) receptor antagonist (propranolol) reduced heart rate (69 +/- 8 to 58 +/- 6 beats min(-1)) and exercise capacity (239 +/- 42 to 209 +/- 31 W; P < 0.05) with arterial lactate reaching 9.4 +/- 3.6 mm. During exercise with propranolol, the increase in a-v lactate difference (to 0.5 +/- 0.5 mm; P < 0.05) was attenuated and the a-v glucose difference and the cerebral metabolic ratio remained at levels similar to those at rest. Together with the previous finding that the cerebral metabolic ratio is unaffected during exercise with administration of the beta(1)-receptor antagonist metropolol, the present results suggest that the cerebral metabolic ratio decreases in response to a beta(2)-receptor mechanism. PMID- 18403424 TI - 2-Aminoethoxydiphenyl borate directly facilitates and indirectly inhibits STIM1 dependent gating of CRAC channels. AB - 2-Aminoethoxydiphenyl borate (2-APB) has emerged as a useful pharmacological tool in the study of store-operated Ca(2+) entry (SOCE). It has been shown to potentiate store-operated Ca(2+) release-activated Ca(2+) (CRAC) currents at low micromolar concentrations and to inhibit them at higher concentrations. Initial experiments with the three CRAC channel subtypes CRACM1, CRACM2 and CRACM3 have indicated that they might be differentially affected by 2-APB. We now present a thorough pharmacological profile of 2-APB and report that it can activate CRACM3 channels in a store-independent manner without the requirement of STIM1, whereas CRACM2 by itself is completely unresponsive to 2-APB and CRACM1 is only very weakly activated. However, when coexpressed with STIM1 and activated via store depletion, CRACM1 and CRACM2 are facilitated at low 2-APB concentrations and inhibited at higher concentrations, while CRACM3 only exhibits potentiated currents. Consistently, the 2-APB-induced CRAC currents exhibit altered selectivities that are characterized by a leftward shift in reversal potential and the emergence of large outward currents that are carried by normally impermeant monovalent cations such as Cs(+) or K(+). These results suggest that 2 APB has agonistic and antagonistic modes of action on CRAC channels, acting at the channel level as a store-independent and direct gating agonist for CRACM3 and a potentiating agonist for CRACM1 and CRACM2 following store-operated and STIM1 dependent activation. The inhibition of CRACM1 channels by high concentrations of 2-APB appears to involve a direct block at the channel level and an additional uncoupling of STIM1 and CRACM1, since the compound reversed the store-dependent multimerization of STIM1. Finally, we demonstrate that single-point mutations of critical amino acids in the selectivity filter of the CRACM1 pore (E106D and E190A) enable 2-APB to gate CRACM1 in a STIM1-independent manner, suggesting that 2-APB facilitates CRAC channels by altering the pore architecture. PMID- 18403425 TI - Modulation of the control of muscle sympathetic nerve activity during incremental leg cycling. AB - We tested the hypotheses that arterial baroreflex (ABR) control over muscle sympathetic nerve activity (MSNA) in humans does not remain constant throughout a bout of leg cycling ranging in intensity from very mild to exhausting. ABR control over MSNA (burst incidence, burst strength and total MSNA) was evaluated by analysing the relationship between beat-to-beat spontaneous variations in diastolic arterial pressure (DAP) and MSNA in 15 healthy subjects at rest and during leg cycling in a seated position at five workloads: very mild (10 W), mild (82 +/- 5.0 W), moderate (126 +/- 10.2 W), heavy (156 +/- 14.3 W), and exhausting (190 +/- 21.2 W). The workload was incremented every 6 min. The linear relationships between DAP and MSNA variables were significantly shifted downward during very mild exercise, but then shifted progressively upward as exercise intensity increased. During heavy and exhausting exercise, moreover, the DAP-MSNA relationships were also significantly shifted rightward from the resting relationship. The sensitivity of ABR control over burst incidence and total MSNA was significantly lower during very mild exercise than during rest, and the sensitivity of the burst incidence control remained lower than the resting level at all higher exercise intensities. By contrast, the sensitivity of the total MSNA control recovered to the resting level during mild and moderate exercise, and was significantly increased during heavy and exhausting exercise (versus rest). We conclude that, in humans, ABR control over MSNA is not uniform throughout a leg cycling exercise protocol in which intensity was varied from very mild to exhausting. We suggest that this non-uniformity of ABR function is one of the mechanisms by which sympathetic and cardiovascular responses are matched to the exercise intensity. PMID- 18403426 TI - Two different forms of long-term potentiation at CA1-subiculum synapses. AB - Distinct functional roles in learning and memory are attributed to certain areas of the hippocampus and the parahippocampal region. The subiculum as a part of the hippocampal formation is the principal target of CA1 pyramidal cell axons and serves as an interface in the information processing between the hippocampus and the neocortex. Subicular pyramidal cells have been classified as bursting and regular firing cells. Here we report fundamental differences in long-term potentiation (LTP) between both cell types. Prolonged high-frequency stimulation induced NMDA receptor-dependent LTP in both cell types. While LTP relied on postsynaptic calcium in regular firing neurons, no increase in postsynaptic calcium was required in bursting cells. Furthermore, paired-pulse facilitation revealed that the site of LTP expression was postsynaptic in regular firing neurons, while presynaptic in burst firing neurons. Our findings on synaptic plasticity in the subiculum indicate that regular firing and bursting cells represent two functional units with distinct physiological roles in processing hippocampal output. PMID- 18403427 TI - Explaining everything? The power and perils of reading Rosenberg. AB - This article examines the writings and teachings of eminent American medical historian Charles E. Rosenberg from the perspective of one of his former graduate students. It examines the appeal of the integrative quality of Rosenberg's historical approach; his attention to imperfect and inconsistent ideology; his use of graphic examples to shock and engage; his preference for continuity over change; his rejection of nostalgia and romanticism; the influence of his teacher Erwin Ackerknecht; and Rosenberg's response to American health policy issues. The article also places Rosenberg within the history of the rise and fall of American social medicine and assesses the potential influence of his work for twenty-first century American medical history and health policy. PMID- 18403428 TI - Charles E. Rosenberg and the multifaceted promise of medical history. AB - Charles E. Rosenberg has had a major influence in defining the history of medicine as a field. However, critics who focus on his leadership or "school" in terms of defined scholarly perspectives, including those of social history and the framing of disease, offer inadequate descriptions of the messages, breadth, and scope of his scholarly work as a whole. Shoehorning the history of medicine into prescribed patterns in order to build a more unitary discipline would weaken rather than strengthen the field and is not in the Rosenberg tradition. PMID- 18403429 TI - Continuity and contingency: the medical-historical world according to Charles E. Rosenberg. PMID- 18403430 TI - ACCORD and ADVANCE: a tale of two studies on the merits of glycaemic control in type 2 diabetic patients. PMID- 18403431 TI - Urinary excretion of fatty acid-binding proteins in idiopathic membranous nephropathy. AB - BACKGROUND: It is suggested that proteinuria contributes to progressive renal failure by inducing tubular cell injury. The site of injury is unknown. Most studies have used markers of proximal tubular cell damage. Fatty acid-binding proteins (FABPs) are intracellular carrier proteins with different expression in the kidney. Liver-type FABP (L-FABP) is found in the cytoplasm of proximal tubules, whereas heart-type FABP (H-FABP) is localized in the distal tubules. We evaluated the urinary excretion of L-FABP and H-FABP in patients with idiopathic membranous nephropathy (iMN). METHODS: We have studied 40 patients (27 males, 13 females) with iMN. The mean age was 48 +/- 15 years, serum creatinine concentration 89 +/- 17 micromol/l and proteinuria 8.9 +/- 5.0 g/24 h. Urinary L FABP and H-FABP were measured by ELISA. Renal failure was defined as an increase in serum creatinine >25% from baseline with a serum creatinine >135 micromol/l or an increase >50% from baseline. Urinary L-FABP excretion was detectable in all but one patient. The median (range) level was 3.29 (0.7-165.6) microg/mmol creatinine (normal <0.38 microg/mmol Cr). Urinary H-FABP was undetectable in nine patients. Median level was 1.53 (0.1-90.5) microg/mmol Cr (normal <0.1 microg/mmol Cr). Both L- and H-FABP correlated with urinary beta2-microglobulin, urinary alpha1-microglobulin and IgG. Urinary H-FABP paralleled L-FABP. RESULTS: After a mean follow-up of 75 +/- 32 months, 16 (40%) patients have reached the predefined end point of renal failure. Both urinary L-FABP and H-FABP predicted renal outcome, with the calculated sensitivity and specificity of 81 and 83% for both. CONCLUSIONS: Urinary L-FABP and urinary H-FABP are increased in patients with iMN. There was a high correlation between L-FABP and H-FABP, suggesting the concurrent development or existence of proximal and distal tubular cell injury. Both L-FABP and H-FABP predicted prognosis in patients with iMN. These markers may be of interest as research tools; however, they are not superior to more conventional marker proteins. PMID- 18403432 TI - Cardiac troponin T predicts occult coronary artery stenosis in patients with chronic kidney disease at the start of renal replacement therapy. AB - BACKGROUND: The high prevalence of asymptomatic coronary artery stenosis (CAS) in chronic kidney disease (CKD) has emerged as an important predictor of outcome. However, diagnostic tools that can identify asymptomatic CAS have not yet been established. We investigated whether asymptomatic patients at the initiation of renal replacement therapy (RRT) could be screened using cardiac troponin T (cTnT) and atherosclerotic surrogate markers such as ankle-brachial blood pressure index (ABPI) and intima-media thickness (IMT). METHODS AND RESULTS: Among 142 patients who were about to start RRT, 60 who were asymptomatic underwent coronary evaluation by multi-slice computed tomography (MSCT) and/or coronary angiography (CAG). CAG diagnosed 35 patients (43.8%) as CAS positive and 27 of them had multi vessel disease. Factors associated with CAS were smoking, elevated cTnT, low ABPI and high IMT. Moreover, the severity of CAS was associated with smoking, cTnT and ABPI. Stepwise logistic regression analyses revealed that cTnT was a powerful predictor of asymptomatic multi-vessel CAS. Receiver operating characteristic analysis documented the usefulness of cTnT as a screening tool with a cut-off point 0.05 ng/ml. The optimal screening tool for multi-vessel CAS was cTnT (sensitivity, 92.6%; 95% CI, 82.7-99.9; specificity, 63.6%; 95% CI, 47.2-80.0). CONCLUSION: We concluded that cTnT should be measured as part of a strategy for detecting asymptomatic CAS, especially multi-vessel disease in patients with CKD at the start of RRT. PMID- 18403433 TI - The long forgotten salt factor and the benefits of using a 5-g-salt-restricted diet in all ESRD patients. PMID- 18403434 TI - Bacterial protease for the treatment of IgA nephropathy. PMID- 18403435 TI - The role for adjunctive treatment to plasma exchange in thrombotic thrombocytopenic purpura. PMID- 18403436 TI - Increased serum advanced glycation end products are associated with impairment in HDL antioxidative capacity in diabetic nephropathy. PMID- 18403437 TI - Hopeful for a second HOPE-2 post hoc analysis. PMID- 18403438 TI - High mean fasting glucose levels independently predict poor outcome and delayed cerebral ischaemia after aneurysmal subarachnoid haemorrhage. AB - BACKGROUND: Hyperglycaemia has been related to poor outcome and delayed cerebral ischaemia (DCI) after aneurysmal subarachnoid haemorrhage (aSAH). OBJECTIVE: This study aimed to assess whether in patients with aSAH, levels of mean fasting glucose within the first week predict poor outcome and DCI better than single admission glucose levels alone. METHODS: Data on non-diabetic patients admitted within 48 h after aSAH with at least two fasting glucose assessments in the first week were retrieved from a prospective database (n = 265). The association of admission glucose or mean fasting glucose, dichotomised at the median levels, with outcome was assessed using logistic regression, and for DCI using Cox regression. To explore whether the association between glucose levels and outcome was mediated by DCI, we adjusted for DCI. RESULTS: The crude and multivariable adjusted odds ratios and 95% confidence intervals for poor outcome were 1.9 (1.1 to 3.2) and 1.6 (0.9 to 2.7) for high admission glucose and 3.5 (2.0 to 6.1) and 2.5 (1.4 to 4.6) for high mean fasting glucose. The crude and adjusted hazard ratios for DCI were 1.7 (1.1 to 2.5) and 1.4 (0.9 to 2.1) for high admission glucose and 2.0 (1.3 to 3.0) and 1.7 (1.1 to 2.7) for high mean fasting glucose. After adjusting for DCI, the odds ratios on poor outcome for high mean fasting glucose decreased only marginally. CONCLUSIONS: Compared with high admission glucose, high mean fasting glucose, representing impaired glucose metabolism, is a better and independent predictor of poor outcome and DCI. DCI is not the key determinant in the relationship between high fasting glucose and poor outcome. PMID- 18403439 TI - Attempted and completed suicides after subthalamic nucleus stimulation for Parkinson's disease. AB - A higher than expected frequency of suicide has been reported among patients undergoing subthalamic nucleus deep brain stimulation (STN DBS) for advanced Parkinson's disease (PD). We conducted a retrospective survey of 200 patients with PD who underwent STN DBS. Two patients (1%) committed suicide and four (2%) attempted suicide, despite clear motor improvements. Suicidal patients did not differ from non-suicidal patients with respect to age, disease duration or preoperative depressive and cognitive status. Suicidal behaviour was associated with postoperative depression and/or altered impulse regulation. Suicidal behaviour is a potential hazard of STN DBS, calling for careful preoperative assessment and close postoperative psychiatric and behavioural follow-up. PMID- 18403441 TI - Interstitial spinal-cord oedema in syringomyelia associated with Chiari type 1 malformations. AB - OBJECT: The pathophysiology of syringomyelia in Chiari type 1 malformations has not been clarified. Oedema-like spinal-cord swelling was recently reported in several pathological conditions, including Chiari type 1 malformations as a pre syrinx state. However, the role of the pre-syrinx state in the development of syringomyelia is unknown. The purpose of this study is to investigate the parenchymal changes of the spinal cord in syringomyelia associated with Chiari type 1 malformations. METHODS: Pre- and postoperative MRI findings in 14 patients who underwent foramen magnum decompression in our institute were reviewed. The analysis was focused on differences in visualisation of the syrinx between T1- and T2-weighted images and abnormal parenchymal signal changes. There were 6 men and 8 women, aged from 6 to 79 years. No patients showed hydrocephalus. RESULTS: Twelve patients had large and expansive syrinx, whereas 2 patients showed small syrinx confined to the centre of the spinal cord. T2-weighted images displayed significantly larger intramedullary abnormal signal areas. Nine patients showed parenchymal hyperintensity areas around the enlarged central canal or base of the posterior white columns adjacent to the syringomyelic cavity. Such parenchymal hyperintensity areas markedly diminished with reduction of the syrinx after surgery and were considered to be interstitial oedema. CONCLUSIONS: From this study, the interstitial oedema of the spinal cord commonly accompanies syringomyelia with Chiari type 1 malformations. Accumulation of the extracellular fluid due to disturbed absorption mechanisms may play an important role in the pathophysiology of syringomyelia associated with Chiari type 1 malformations. PMID- 18403440 TI - Deep brain stimulation activation volumes and their association with neurophysiological mapping and therapeutic outcomes. AB - OBJECTIVE: Despite the clinical success of deep brain stimulation (DBS) for the treatment of Parkinson's disease (PD), little is known about the electrical spread of the stimulation. The primary goal of this study was to integrate neuroimaging, neurophysiology and neurostimulation data sets from 10 patients with PD, unilaterally implanted with subthalamic nucleus (STN) DBS electrodes, to identify the theoretical volume of tissue activated (VTA) by clinically defined therapeutic stimulation parameters. METHODS: Each patient specific model was created with a series of five steps: (1) definition of the neurosurgical stereotactic coordinate system within the context of preoperative imaging data; (2) entry of intraoperative microelectrode recording locations from neurophysiologically defined thalamic, subthalamic and substantia nigra neurons into the context of the imaging data; (3) fitting a three dimensional brain atlas to the neuroanatomy and neurophysiology of the patient; (4) positioning the DBS electrode in the documented stereotactic location, verified by postoperative imaging data; and (5) calculation of the VTA using a diffusion tensor based finite element neurostimulation model. RESULTS: The patient specific models show that therapeutic benefit was achieved with direct stimulation of a wide range of anatomical structures in the subthalamic region. Interestingly, of the five patients exhibiting a greater than 40% improvement in their Unified PD Rating Scale (UPDRS), all but one had the majority of their VTA outside the atlas defined borders of the STN. Furthermore, of the five patients with less than 40% UPDRS improvement, all but one had the majority of their VTA inside the STN. CONCLUSIONS: Our results are consistent with previous studies suggesting that therapeutic benefit is associated with electrode contacts near the dorsal border of the STN, and provide quantitative estimates of the electrical spread of the stimulation in a clinically relevant context. PMID- 18403442 TI - Simple is beautiful: a straightforward approach to improve the delineation of true and false positives in PSI-BLAST searches. AB - MOTIVATION: The deluge of biological information from different genomic initiatives and the rapid advancement in biotechnologies have made bioinformatics tools an integral part of modern biology. Among the widely used sequence alignment tools, BLAST and PSI-BLAST are arguably the most popular. PSI-BLAST, which uses an iterative profile position specific score matrix (PSSM)-based search strategy, is more sensitive than BLAST in detecting weak homologies, thus making it suitable for remote homolog detection. Many refinements have been made to improve PSI-BLAST, and its computational efficiency and high specificity have been much touted. Nevertheless, corruption of its profile via the incorporation of false positive sequences remains a major challenge. RESULTS: We have developed a simple and elegant approach to resolve the problem of model corruption in PSI BLAST searches. We hypothesized that combining results from the first (least corrupted) profile with results from later (most sensitive) iterations of PSI BLAST provides a better discriminator for true and false hits. Accordingly, we have derived a formula that utilizes the E-values from these two PSI-BLAST iterations to obtain a figure of merit for rank-ordering the hits. Our verification results based on a 'gold-standard' test set indicate that this figure of merit does indeed delineate true positives from false positives better than PSI-BLAST E-values. Perhaps what is most notable about this strategy is that it is simple and straightforward to implement. PMID- 18403443 TI - Interleukin-4 activates large-conductance, calcium-activated potassium (BKCa) channels in human airway smooth muscle cells. AB - Large-conductance, calcium-activated potassium (BK(Ca)) channels are regulated by voltage and near-membrane calcium concentrations and are determinants of membrane potential and excitability in airway smooth muscle cells. Since the T helper-2 (Th2) cytokine, interleukin (IL)-4, is an important mediator of airway inflammation, we investigated whether IL-4 rapidly regulated BK(Ca) activity in normal airway smooth muscle cells. On-cell voltage clamp recordings were made on subconfluent, cultured human bronchial smooth muscle cells (HBSMC). Interleukin-4 (50 ng ml(-1)), IL-13 (50 ng ml(-1)) or histamine (10 microm) was added to the bath during the recordings. Immunofluorescence studies with selective antibodies against the alpha and beta1 subunits of BK(Ca) were also performed. Both approaches demonstrated that HBSMC membranes contained large-conductance channels (>200 pS) with both calcium and voltage sensitivity, all of which is characteristic of the BK(Ca) channel. Histamine caused a rapid increase in channel activity, as expected. A new finding was that perfusion with IL-4 stimulated rapid, large increases in BK(Ca) channel activity (77.2 +/- 63.3-fold increase, P < 0.05, n = 18). This large potentiation depended on the presence of external calcium. In contrast, IL-13 (50 ng ml(-1)) had little effect on BK(Ca) channel activity, but inhibited the effect of IL-4. Thus, HBSMC contain functional BK(Ca) channels whose activity is rapidly potentiated by the cytokine, IL-4, but not by IL-13. These findings are consistent with a model in which IL-4 rapidly increases near-membrane calcium concentrations to regulate BK(Ca) activity. PMID- 18403444 TI - Effects of chemoreflexes on hyperthermic hyperventilation and cerebral blood velocity in resting heated humans. AB - We tested the hypothesis that hyperthermic hyperventilation in part reflects enhanced chemoreceptor ventilatory O(2) drive, and that the resultant hypocapnia attenuates ventilatory responses and/or middle cerebral artery mean blood velocity (MCAV(mean)) in resting humans. Eleven healthy subjects were passively heated for 50-80 min, causing oesophageal temperature (T(oes)) to increase by 1.6 degrees C. During heating, minute ventilation increased (P < 0.05), while end tidal CO(2) pressure (P(ET,CO(2))) and MCAV(mean) declined. A hyperoxia test in which three breaths of hyperoxic air were inspired was performed once before heating and three times during the heating. When we observed hypocapnia (P(ET,CO(2)) below 40 mmHg), P(ET,CO(2)) was restored to the eucapnic level by adding 100% CO(2) to the inspired air immediately before the last two tests. Minute ventilation was significantly reduced by hyperoxia, and that reduction gradually increased with increasing T(oes). However, the percentage decrease in from the normoxic level was small (20-29%) and unchanged during heating. When P(ET,CO(2)) was restored to eucapnic levels, was unchanged, but MCAV(mean) was partly restored to the level seen prior to heating (28.1% restoration at T(oes) 37.6 degrees C and 38.1% restoration at T(oes) 38.0 degrees C). These findings suggest that although hyperthermia increases chemoreceptor ventilatory O(2) drive in resting humans, the relative contribution of the chemoreceptor ventilatory O(2) drive to hyperthermic hyperventilation is small ( approximately 20%) and unaffected by increasing core temperature. Moreover, hypocapnia induced by hyperthermic hyperventilation reduces cerebral blood flow but not ventilatory responses. PMID- 18403446 TI - Increased ventilation does not impair maximal voluntary contractions of the elbow flexors. AB - Exercise performance is impaired by increased respiratory work, yet the mechanism for this is unclear. This experiment assessed whether neural drive to an exercising muscle was affected by cortically driven increases in ventilation. On each of 5 days, eight subjects completed a 2-min maximal voluntary contraction (MVC) of the elbow flexor muscles, followed by 4 min of recovery, while transcranial magnetic stimulation tested for suboptimal neural drive to the muscle. On 1 day, subjects breathed without instructions under normocapnia. During the 2-min MVC, ventilation was approximately 3.5 times that at rest. On another day, subjects breathed without instruction under hypercapnia. During the 2-min MVC, ventilation was approximately 1.5 times that on the normocapnic day. On another 2 days under normocapnia, subjects voluntarily matched their breathing to the uninstructed breathing under normocapnia and hypercapnia using target feedback of the rate and inspiratory volume. On a fifth day under normocapnia, the volume feedback was set to each subject's vital capacity. On this day, ventilation during the 2-min MVC was approximately twice that on the uninstructed normocapnic day (or approximately 7 times rest). The experimental manipulations succeeded in producing voluntary and involuntary hyperpnea. However, maximal voluntary force, fatigue and voluntary activation of the elbow flexor muscles were unaffected by cortically or chemically driven increases in ventilation. Results suggest that any effects of increased respiratory work on limb exercise performance are not due to a failure to drive both muscle groups optimally. PMID- 18403445 TI - Role of TRPV1 channels in renal haemodynamics and function in Dahl salt-sensitive hypertensive rats. AB - This study tests the hypothesis that dysfunction of transient receptor potential vanilloid type 1 (TRPV1) channels occurs and contributes to the decrease in the glomerular filtration rate (GFR) and sodium/water excretion in Dahl salt sensitive hypertensive rats. Recirculating Krebs-Henseleit buffer added with inulin was perfused at a constant flow in the isolated kidneys of Dahl salt sensitive (DS) or Dahl salt-resistant (DR) rats fed a high-salt (HS) or low-salt (LS) diet for 3 weeks. Perfusion pressures (PP) were pre-adjusted to three levels ( approximately 100, approximately 150 or approximately 190 mmHg) with or without phenylephrine. Capsaicin, a selective TRPV1 agonist, in the presence or absence of capsazepine, a selective TRPV1 antagonist, was perfused. Basal GFR, urine flow rate (UFR) and Na(+) excretion (U(Na)V) were significantly lower in DS-HS than in DR-HS, DS-LS and DR-LS rats. Capsaicin caused pressure-dependent decreases in PP and increases in GFR, UFR and U(Na)V in all groups, with less magnitude of decreases in PP and increases in GFR, UFR and U(Na)V in DS-HS than in DR-HS, DS LS and DR-LS rats. Capsazepine completely blocked the effect of capsaicin on PP, GFR, UFR and U(Na)V in all groups. Thus, these results show that TRPV1 function is impaired in the kidney of DS rats fed a high-salt diet, which may contribute to the decrease in GFR and renal excretory function in DS rats in the face of salt challenge. PMID- 18403447 TI - Unilateral practice of a ballistic movement causes bilateral increases in performance and corticospinal excitability. AB - It has long been known that practicing a task with one limb can result in performance improvements with the opposite, untrained limb. Hypotheses to account for cross-limb transfer of performance state that the effect is mediated either by neural adaptations in higher order control centers that are accessible to both limbs, or that there is a "spillover" of neural drive to the opposite hemisphere that results in bilateral adaptation. Here we address these hypotheses by assessing performance and corticospinal excitability in both hands after unilateral practice of a ballistic finger movement. Participants (n = 9) completed 300 practice trials of a ballistic task with the right hand, the aim of which was to maximize the peak abduction acceleration of the index finger. Practice caused a 140% improvement in right-hand performance and an 82% improvement for the untrained left hand. There were bilateral increases in the amplitude of responses to transcranial magnetic stimulation, but increased corticospinal excitability was not correlated with improved performance. There were no significant changes in corticospinal excitability or task performance for a control group that did not train (n = 9), indicating that performance testing for the left hand alone did not induce performance or corticospinal effects. Although the data do not provide conclusive evidence whether increased corticospinal excitability in the untrained hand is causally related to the cross transfer of ballistic performance, the finding that ballistic practice can induce bilateral corticospinal adaptations may have important clinical implications for movement rehabilitation. PMID- 18403448 TI - Effect of exercise on in vitro immune function: a 12-month randomized, controlled trial among postmenopausal women. AB - Cross-sectional studies suggest that moderate physical activity is associated with enhanced resting immune function; however, few randomized controlled trials have investigated this link. We investigated the effect of 12-mo aerobic exercise, relative to stretching control, on in vitro immune function in a randomized, controlled trial of 115 postmenopausal, overweight, or obese sedentary women, aged 50-75 yr. The exercise goal was > or =45 min/day, 5 days/wk. Control women participated in 1 day/wk stretching classes. Immune markers (natural killer cell cytotoxicity, T-lymphocyte proliferation, immune cell counts and phenotypes, and serum immunoglobulins) were assessed at baseline, 3 mo, and 12 mo under strict blood-draw criteria. General estimation equations evaluated intervention effects at 3 and 12 mo, controlling for baseline. Of the 115 women who began the trial, blood samples were available from 109 at 3 mo (95%) and 108 at 12 mo (94%). From baseline to 12 mo, the exercise group participated in 87% of the prescribed physical activity minutes per week and increased maximal O(2) uptake by 13.8%; controls experienced no change in fitness. The main outcomes, natural killer cell cytotoxicity and T-lymphocyte proliferation, did not differ between groups at 3 and 12 mo. Secondary outcome and subgroup (e.g., stratification by baseline categories of body mass index, immune status, C-reactive protein, and age) analyses did not show any clear patterns of association. This 12-mo randomized, controlled trial showed no effect of aerobic exercise on in vitro immune function, despite excellent retention, high adherence, and demonstrable efficacy of the exercise intervention. PMID- 18403449 TI - Effects of chronic electrical stimulation on paralyzed expiratory muscles. AB - Following spinal cord injury, the expiratory muscles develop significant disuse atrophy characterized by reductions in their weight, fiber cross-sectional area, and force-generating capacity. We determined the extent to which these physiological alterations can be prevented with electrical stimulation. Because a critical function of the expiratory muscles is cough generation, an important goal was the maintenance of maximal force production. In a cat model of spinal cord injury, short periods of high-frequency lower thoracic electrical spinal cord stimulation (SCS) at the T(10) level (50 Hz, 15 min, twice/day, 5 days/wk) were initiated 2 wk following spinalization and continued for a 6-mo period. Airway pressure (P)-generating capacity was determined by SCS. Five acute, spinalized animals served as controls. Compared with controls, initial P fell from 43.9 +/- 1.0 to 41.8 +/- 0.7 cmH(2)O (not significant) in the chronic animals. There were small reductions in the weight of the external oblique, internal oblique, transverses abdominis, internal intercostal, and rectus abdominis muscles (not significant for each). There were no significant changes in the population of fast muscle fibers. Because prior studies (Kowalski KE, Romaniuk JR, DiMarco AF. J Appl Physiol 102: 1422-1428, 2007) have demonstrated significant atrophy following spinalization in this model, these results indicate that expiratory muscle atrophy can be prevented by the application of short periods of daily high-frequency stimulation. Because the frequency of stimulation is similar to the expected pattern of clinical use for cough generation, the daily application of electrical stimulation could potentially serve the dual purpose of maintenance of expiratory muscle function and airway clearance. PMID- 18403450 TI - Impact of intermittent hypoxia on long-term facilitation of minute ventilation and heart rate variability in men and women: do sex differences exist? AB - Following exposure to intermittent hypoxia, respiratory motor activity and sympathetic nervous system activity may persist above baseline levels for over an hour. The present investigation was designed to determine whether sustained increases in minute ventilation and sympathovagal (S/V) balance, in addition to sustained depression of parasympathetic nervous system activity (PNSA), were greater in men compared with women following exposure to intermittent hypoxia. Fifteen healthy men and women matched for age, race, and body mass index were exposed to eight 4-min episodes of hypoxia during sustained hypercapnia followed by a 15-min end-recovery period. The magnitude of the increase in minute ventilation during the end-recovery period, compared with baseline, was similar in men and women (men, 1.52 +/- 0.03; women, 1.57 +/- 0.02 fraction of baseline; P < 0.0001). In contrast, depression of PNSA and increases in S/V balance were evident during the end-recovery period, compared with baseline, in men (PNSA, 0.66 +/- 0.06 fraction of baseline, P < 0.0001; S/V balance, 2.8 +/- 0.7 fraction of baseline, P < 0.03) but not in women (PNSA, 1.27 +/- 0.19 fraction of baseline, P = 0.3; S/V balance, 1.8 +/- 0.6 fraction of baseline, P = 0.2). We conclude that a sustained increase in minute ventilation, which is indicative of long-term facilitation, is evident in both men and women following exposure to intermittent hypoxia and that this response is independent of sex. In contrast, sustained alterations in autonomic nervous system activity were evident in men but not in women. PMID- 18403451 TI - Upper airway neuromuscular compensation during sleep is defective in obstructive sleep apnea. AB - Obstructive sleep apnea is the result of repeated episodes of upper airway obstruction during sleep. Recent evidence indicates that alterations in upper airway anatomy and disturbances in neuromuscular control both play a role in the pathogenesis of obstructive sleep apnea. We hypothesized that subjects without sleep apnea are more capable of mounting vigorous neuromuscular responses to upper airway obstruction than subjects with sleep apnea. To address this hypothesis we lowered nasal pressure to induce upper airway obstruction to the verge of periodic obstructive hypopneas (cycling threshold). Ten patients with obstructive sleep apnea and nine weight-, age-, and sex-matched controls were studied during sleep. Responses in genioglossal electromyography (EMG(GG)) activity (tonic, peak phasic, and phasic EMG(GG)), maximal inspiratory airflow (V(I)max), and pharyngeal transmural pressure (P(TM)) were assessed during similar degrees of sustained conditions of upper airway obstruction and compared with those obtained at a similar nasal pressure under transient conditions. Control compared with sleep apnea subjects demonstrated greater EMG(GG), V(I)max, and P(TM) responses at comparable levels of mechanical and ventilatory stimuli at the cycling threshold, during sustained compared with transient periods of upper airway obstruction. Furthermore, the increases in EMG(GG) activity in control compared with sleep apnea subjects were observed in the tonic but not the phasic component of the EMG response. We conclude that sustained periods of upper airway obstruction induce greater increases in tonic EMG(GG), V(I)max, and P(TM) in control subjects. Our findings suggest that neuromuscular responses protect individuals without sleep apnea from developing upper airway obstruction during sleep. PMID- 18403453 TI - Application of cross-sectional time series modeling for the prediction of energy expenditure from heart rate and accelerometry. AB - Accurate estimation of energy expenditure (EE) in children and adolescents is required for a better understanding of physiological, behavioral, and environmental factors affecting energy balance. Cross-sectional time series (CSTS) models, which account for correlation structure of repeated observations on the same individual, may be advantageous for prediction of EE. CSTS models for prediction of minute-by-minute EE and, hence, total EE (TEE) from heart rate (HR), physical activity (PA) measured by accelerometry, and observable subject variables were developed in 109 children and adolescents by use of Actiheart and 24-h room respiration calorimetry. CSTS models based on HR, PA, time-invariant covariates, and interactions were developed. These dynamic models involve lagged and lead values of HR and lagged values of PA for better description of the series of minute-by-minute EE. CSTS models with random intercepts and random slopes were investigated. For comparison, likelihood ratio tests were used. Log likelihood increased substantially when random slopes for HR and PA were added. The population-specific model uses HR and 1- and 2-min lagged and lead values of HR, HR(2), and PA and 1- and 2-min lagged values of PA, PA(2), age, age(2), sex, weight, height, minimum HR, sitting HR, HR x height, HR x weight, HR x age, PA x weight, and PA x sex interactions (P < 0.001). Prediction error for TEE was 0.9 +/- 10.3% (mean +/- SD). Errors were not correlated with age, weight, height, or body mass index. CSTS modeling provides a useful predictive model for EE and, hence, TEE in children and adolescents on the basis of HR and PA and other observable explanatory subject characteristics of age, sex, weight, and height. PMID- 18403452 TI - Key aspects of phrenic motoneuron and diaphragm muscle development during the perinatal period. AB - At the time of birth, respiratory muscles must be activated to sustain ventilation. The perinatal development of respiratory motor units (comprising an individual motoneuron and the muscle fibers it innervates) shows remarkable features that enable mammals to transition from in utero conditions to the air environment in which the remainder of their life will occur. In addition, significant postnatal maturation is necessary to provide for the range of motor behaviors necessary during breathing, swallowing, and speech. As the main inspiratory muscle, the diaphragm muscle (and the phrenic motoneurons that innervate it) plays a key role in accomplishing these behaviors. Considerable diversity exists across diaphragm motor units, but the determinant factors for this diversity are unknown. In recent years, the mechanisms underlying the development of respiratory motor units have received great attention, and this knowledge may provide the opportunity to design appropriate interventions for the treatment of respiratory disease not only in the perinatal period but likely also in the adult. PMID- 18403454 TI - Exercise training enhances in vivo tuberculosis purified protein derivative response in the elderly. AB - We investigated the effect of 25 wk of exercise training on in vivo immune measures that depend on T helper 1 (Th1) and T helper 2 (Th2) immune responses in the elderly as a substudy of a randomized controlled trial to investigate health benefits of regular exercise training for the elderly. Sixty-five healthy elderly volunteers were randomly assigned to either an exercise training group (n = 32) or a sedentary control group (n = 33). The area of skin reaction to purified protein derivative (PPD) of tuberculin that depends on Th1 activation and the concentrations of serum IgG subclasses and IgE were evaluated before and after 25 wk intervention. All participants completed 25 wk of training. Thirty-one participants of the exercise group and all control group underwent immunological analyses, but only 30 from the exercise group and 21 from the control group had the PPD skin reaction assessment. Repeated-measures ANOVA revealed a significant interaction between time and exercise intervention, which appeared as an enhanced skin reaction to tuberculin PPD (P < 0.05) and a reduced serum IgG4 concentration, the production of which depends on Th2-dependent class switching (P < 0.05), in the exercise group after 25 wk. No immune variables changed in the control group. These result supports the hypothesis that exercise training favors in vivo Th1 immune response in elderly persons. PMID- 18403455 TI - Effects of long-term immobilization and recovery on human triceps surae and collagen turnover in the Achilles tendon in patients with healing ankle fracture. AB - The aim of the present study was to analyze how human tendon connective tissue responds to an approximately 7-wk period of immobilization and a remobilization period of a similar length, in patients with unilateral ankle fracture, which is currently unknown. Calf muscle cross-sectional area (CSA) decreased by 15% (5,316 to 4,517 mm2) and strength by 54% (239 to 110 N.m) in the immobilized leg after 7 wk. During the 7-wk remobilization, the CSA increased by 9% (to 4,943 mm2) and strength by 37% (to 176 Nm). Achilles tendon CSA did not change significantly during either immobilization or remobilization. Local collagen turnover was measured as the peritendinous concentrations of NH2-terminal propeptide of type I collagen (PINP) and COOH-terminal telopeptide region of type I collagen (ICTP), markers thought to be indexes of type I collagen synthesis and degradation, respectively. Both markers were increased (PINP: 257 vs. 56 ng/ml; ICTP: 9.8 vs. 2.1 microg/l) in the immobilized leg compared with the control leg after the 7 wk of immobilization, and levels decreased again in the immobilized leg during the recovery period (PINP: 103 vs. 44 ng/ml; ICTP: 4.2 vs. 1.9 microg/l). A significant reduction in calf muscle CSA and strength was found in relation to 7 wk of immobilization. Immobilization increased both collagen synthesis and degradation in tendon near tissue. However, it cannot be excluded that the facture of the ankle in close proximity could have affected these data. Remobilization increased muscle size and strength and tendon synthesis and degradation decreased to baseline levels. These dynamic changes in tendon connective tissue turnover were not associated with macroscopic changes in tendon size. PMID- 18403456 TI - Physical exercise improves plasmatic levels of IL-10, left ventricular end diastolic pressure, and muscle lipid peroxidation in chronic heart failure rats. AB - Chronic heart failure (CHF) is characterized by left ventricular dysfunction, resulting in hemodynamic changes, sustained inflammatory state, as well as increase in oxidative stress. Physical exercise has been described as an important nonpharmacological procedure in the treatment of CHF, contributing to the improvement of the clinical outcomes in this disease. This study evaluated the effects of physical training on hemodynamics, muscle lipid peroxidation, and plasmatic levels of IL-10 in CHF rats. The left coronary artery was ligated to induce CHF, or sham operation was performed in control groups. Rats were assigned to one of four groups: trained CHF (T-CHF, n = 10), sedentary CHF (S-CHF, n = 10), trained sham (T-Sham, n = 10), or sedentary sham (S-Sham, n = 10). Trained animals had carried out a swimming protocol, 60 min/day, 5 days/wk, during 8 wk, whereas sedentary animals remained without training. Eight weeks of physical training promoted an improvement of diastolic function represented by a reduction of the left ventricular end-diastolic pressure in the T-CHF group compared with the S-CHF group (P < 0.05). Lipid peroxidation evaluated in gastrocnemius muscle using thiobarbituric acid reactive substance assay was higher in the S-CHF group compared with all other groups (P < 0.05). However, there were no differences between T-CHF compared with S-Sham and T-Sham groups. The plasmatic levels of IL 10 were lower in the S-CHF group compared with all other groups (P < 0.05). These findings demonstrate that regular physical training using a swimming protocol, with duration of 8 wk, improves the cardiac function and the anti-inflammatory response and reduces muscle cellular damage. PMID- 18403457 TI - Physical functioning before and after total hip arthroplasty: perception and performance. AB - BACKGROUND AND PURPOSE: Self-report and performance-based measures of physical functioning in people before and after total hip arthroplasty seem to present different information. The relationship between these different measures is not well understood, and little information is available about changes in this relationship over time. The aims of this study were: (1) to determine the relationship between self-report and performance-based measures of physical functioning before and after total hip arthroplasty, (2) to assess the influence of pain on the relationship, and (3) to determine whether the relationship changes over time. SUBJECTS AND METHODS: Seventy-five subjects admitted for total hip arthroplasty were included and examined before and 6 and 26 weeks after surgery. The relationships between the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) physical functioning subscale and walking speed and gait variability were examined by use of generalized estimating equations, which included interactions with time and the WOMAC pain subscale. RESULTS: The relationship between self-report and performance-based measures of physical functioning was poor. Pain appeared to have a considerable influence on self reported physical functioning. The relationship did not appear to change over time. DISCUSSION AND CONCLUSION: The influence of pain on self-reported physical functioning serves as an explanation for the poor relationship between self reported and performance-based physical functioning. When using a self-report measure such as the WOMAC, one should realize that it does not seem to assess the separate constructs--physical functioning and pain--that are claimed to be measured. PMID- 18403458 TI - Effect of statins on serum apolipoprotein j and paraoxonase-1 levels in patients with ischemic heart disease undergoing coronary angiography. AB - It has been proposed that apolipoprotein J (apo J) and paraoxonase-1 (PON1) correlate with the extent and severity of ischemic heart disease (IHD). This article compares apo J and PON1 serum concentrations, PON1 activity, and the apo J/PON1 ratio in 138 IHD patients (64 statins users and 74 statin nonusers) referred for angiography and possible percutaneous coronary intervention. The effect of statin treatment on apo J and PON1 concentrations, PON1 activity, and the degree of coronary artery stenosis were evaluated. In both groups, apo J levels were increased, whereas PON1 concentration and activity decreased. IHD patients on statins had significantly lower apo J concentration and higher PON1 concentration and activity. Patients on statins had less coronary artery stenosis. High apo J levels, low PON1 levels, low PON1 activity, and a high apo J/PON1 ratio were associated with IHD. Statin treatment reverses these changes, probably by multiple beneficial actions. PMID- 18403459 TI - Intracoronary autologous bone marrow-derived mononuclear cell transplantation improves coronary collateral vessel formation and recruitment capacity in patients with ischemic cardiomyopathy: a combined hemodynamic and scintigraphic approach. AB - This study investigated the effects of intracoronary autologous bone marrow derived mononuclear cell (BMC) transplantation on coronary microcirculation. Fifteen patients with ischemic cardiomyopathy were treated by intracoronary infusion of BMCs via the patent infarct-related artery. The thermodilution derived coronary flow reserve, index of microvascular resistance, pressure derived collateral flow index, and coronary wedge pressure were measured at baseline and at 6 months. Successive balloon inflations during BMC transplantation were performed to observe the recruitment in pressure-derived collateral flow index and coronary wedge pressure, and the percentage changes between baseline and 6 months were calculated. The mean (SD) coronary flow reserve increased from 1.3 (0.4) to 2.1 (0.5), and the mean (SD) index of microvascular resistance decreased from 44.9 (24.4) to 21.2 (14.1) (P = .001 for both). The mean (SD) improvement in pressure-derived collateral flow index (from 0.14 [0.05] to 0.22 [0.08]) was also statistically significant (P = .001). Similarly, the percentage improvements in pressure-derived collateral flow index and coronary wedge pressure were statistically significant (P = .01 for both). The percentage improvement in perfusion assessed by single-photon emission computed tomography strongly correlated with the percentage changes in pressure derived collateral flow index (r = 0.88, P = .001) and coronary wedge pressure (r = 0.69, P = .01). These results demonstrate for the first time (to our knowledge) that intracoronary autologous BMC transplantation improves coronary collateral vessel formation and recruitment capacity in human subjects. PMID- 18403460 TI - Transient left ventricular apical ballooning syndrome: the first series in Taiwanese patients. AB - Previously thought as exclusive in Japanese patients, cases of transient left ventricular apical ballooning from other countries have also been reported. The cause remains unknown. From January 1997 to December 2005, 25 patients presenting with signs and symptoms of acute myocardial infarction with normal coronary arteries were analyzed. In all, 10 patients fulfilled all the criteria for transient left ventricular apical ballooning. In all, 6 patients had chest pain and diaphoresis, 5 patients had ST segment elevation, 7 had T wave inversions, and 5 had QT prolongation; 6 patients had normal coronary arteries and 4 had insignificant stenosis. In all, 2 patients died of sepsis, whereas the rest recovered. This is the first series in Taiwanese patients. Our series showed male preponderance, and most patients recovered with supportive treatment. Without any delineating preangiographic feature differentiating it from acute myocardial infarction, any patient should be treated as a case of myocardial infarction until proven otherwise. PMID- 18403461 TI - Treatment of end-stage "trash feet" with the end-diastolic pneumatic boot. AB - This study reassessed the clinical effect of Circulator Boot (CB) therapy in patients with cholesterol embolization syndrome (CES) of the lower extremities. The medical records were reviewed of 27 patients consecutively referred to the Bryn Mawr Wound Care and Vascular Center with CES who had not responded to previous therapies. All patients with CES referred from January 1, 1997, to September 19, 2005, were followed up and included in the study. The alternate therapy offered for most patients at the time of referral was limb amputation. The median age of the patients was 65 years (age range, 46-84 years) at the time of diagnosis. Healing of CES was observed after a median interval of 11 months (range, 3-32 months) following the initiation of CB therapy. The total number of legs treated was 41. Of 41 legs, 33 (81%) were totally healed, 6 (15%) improved, and 2 (5%) were amputated. After an initial period of improvement, one patient died a month later of causes unrelated to CES or CB therapy. Another patient improved and discontinued treatment before he was totally healed. Cholesterol embolization syndrome is seen predominantly in patients following cardiac or vascular procedures but may occur spontaneously. The CB seems to be the only effective noninvasive therapy for CES. Early initiation of therapy is essential to minimize tissue loss and patient discomfort. PMID- 18403462 TI - Exposure to passive smoking: a test to predict endothelial dysfunction and atherosclerotic lesions. AB - Acute exposure to environmental tobacco smoke (ETS) is considered to adversely influence atherogenesis. The aim of this article was to assess whether brachial ultrasonography in subjects with endothelial dysfunction after ETS exposure is associated with atherosclerotic lesions. Never smoker healthy volunteers (n = 18) and subjects with a previous myocardial infarction (MI; n = 10) were studied. Healthy volunteers were 12 men (66%) and 6 women (34%) with a mean age of 34 +/- 9 years. Post-MI subjects were men with a mean age of 53.8 +/- 4.8 years. After assessing endothelial function (by brachial ultrasonography) at rest, study subjects underwent brachial ultrasonography twice: in a smoke-free environment and then in the same environment polluted by cigarette combustion (35 ppm carbon monoxide concentration). Carboxyhemoglobin concentration was measured before and after ETS exposure. Baseline brachial-artery diameter, diameter during reactive hyperemia, and diameter after sublingual nitroglycerin (GTN) administration (endothelium-independent vasodilator) were measured at rest and in both smoke free and smoking environments. Each study subject acted as their own control. No comparison was made between the two groups. A strong correlation between ETS exposure and endothelial dysfunction was observed in both groups. Post-MI subjects also showed endothelium-independent vasodilation worsening, which is usually due to arterial wall alterations. After ETS exposure, mean flow-mediated vasodilation after GTN was significantly (P < .01) reduced only in post-MI subjects (P < .01). Carboxyhemoglobin concentration increased in both groups (P < .01). ETS exposure may be an effective test to identify endothelial dysfunction and arterial wall alterations by using brachial ultrasonography. PMID- 18403463 TI - Endovascular embolization of a giant renal artery aneurysm with preservation of renal parenchyma. AB - Renal artery aneurysm is a rare condition that has an unclear etiology. Although some patients present with symptoms of hypertension, pain, hematuria, or rupture, the majority are asymptomatic. Traditional surgical repair of renal artery aneurysms is often complex and may require ex vivo repair and reimplantation of the kidney if branch vessels are involved. Very large aneurysms made require nephrectomy. More recently, reports have described endovascular approaches to renal artery aneurysms, including coil embolization and stent graft coverage. This report describes successful endovascular treatment of a 10-cm renal artery aneurysm with preservation of renal mass. PMID- 18403464 TI - Coronary steal due to bilateral internal mammary artery--pulmonary artery fistulas: a rare cause of chest pain after coronary artery bypass grafting. AB - A 54-year-old man with a history of coronary artery bypass grafting (CABG) presented with chest pain and was found to have non-ST-segment elevation myocardial infarction. Left heart catheterization with coronary angiography demonstrated 100% occlusion of the right internal mammary artery (IMA) to the right coronary artery graft in its midsegment and a patent left IMA to the left anterior descending graft. An unusually large extensive fistulous collateral formation was observed between the right IMA and the left IMA to the pulmonary arterial system, causing left to right shunting. His angina was attributed to substantial coronary steal caused by the shunt. The patient refused any further intervention or surgery and opted for medical treatment. As a complication of CABG, IMA to pulmonary artery (PA) fistulas are rare. Thus far, more than 20 cases have been reported; most have been unilateral. This is the second reported case to date of bilateral IMA-PA fistula formation after CABG. An IMA-PA fistula should be considered in the differential diagnosis of patients presenting with chest pain after CABG and can be diagnosed by selective angiography of IMA grafts. PMID- 18403465 TI - Central venous thrombosis in Behcet's disease. AB - Behcet's disease is a systemic inflammatory disorder characterized by vasculitis, which affects all types and sizes of blood vessels. A thrombotic tendency with a predilection for the venous side of the vasculature is well known for Behcet's disease. However, central vein involvement is a rare complication. The authors report an unusual case of complete thrombosis of the supraazygos superior vena cava, brachiocephalic veins, as well as right internal jugular vein in a patient who is a young woman with Behcet's disease. PMID- 18403467 TI - Guest editorial. Papers from the 2007 Midwestern Vascular Surgery Society Annual Meeting. PMID- 18403466 TI - Myocardial infarction with distal embolization associated with myocardial bridging in a postoperative patient: a case report. AB - Myocardial bridging is associated with myocardial infarction (MI) on rare occasions. The mechanism by which this occurs is unclear; vessel occlusion in this setting may be due to superimposed thrombosis or vasospasm. An unusual case is presented of postoperative MI from thrombosis associated with myocardial bridging and intravascular volume depletion, with subsequent distal embolization. Endothelial injury as a result of myocardial bridging may predispose to thrombus formation. To avoid MI, adequate fluid hydration is recommended during the perioperative period in patients with myocardial bridging. PMID- 18403468 TI - Commentary on "Antiplatelet therapy for vascular interventions". PMID- 18403469 TI - Commentary on "The use of stem cells in the treatment of inoperable limb ischemia". PMID- 18403470 TI - Sclerotherapy: a truly minimally invasive technique. AB - Foam sclerotherapy offers a treatment strategy with great potential. Recently, general and vascular surgery have become less invasive; so too, has the treatment of venous disorders. Sclerosants cause irreversible damage to the vascular endothelium by disrupting cell membranes resulting in sustained vasospasm and denudation of the venous monolayer. Prospective randomized outcome data support the hypothesis that foam sclerotherapy is superior to liquid sclerotherapy. All published reports of varicose vein treatment with foam describe efficacy in terms of immediate and primary venous occlusion of better than 80%. Severe complications of foam sclerotherapy are rare. Recurrent varices are in the 10% to 20% range. Use of foam sclerotherapy in our experience has proven to be effective, essentially pain-free, and durable in the short term. The treatment is quick, efficient, and cheap. PMID- 18403471 TI - Commentary on "Endovascular management of deep venous thrombosis". PMID- 18403472 TI - Commentary: Karthik s, et Al. Cryoplasty for arterial restenosis. Eur j vasc endovasc surg. 2007;33:40-43. PMID- 18403473 TI - Commentary: Miyamoto K, Nishigami K, Nagaya N, et al. Unblinded pilot study of autologous transplantation of bone marrow mononuclear cells in patients with thromboangiitis obliterans. Circulation. 2006;114: 2679-2684. PMID- 18403474 TI - Commentary: Everett BM, et al. Prevalence of heparin/platelet factor 4 antibodies before and after cardiac surgery. Ann Thorac Surg. 2007;83:592-597. PMID- 18403475 TI - Imaging mass spectrometry reveals unique protein profiles during embryo implantation. AB - A reciprocal interaction between the implantation-competent blastocyst and receptive uterus is an absolute requirement for implantation, a process crucial for pregnancy success. A comprehensive understanding of this interaction has yet to be realized. One major difficulty in clearly defining this discourse is the complexity of the implantation process involving heterogeneous cell types of both the uterus and blastocyst, each endowed with unique molecular signatures that show dynamic changes during the course of pregnancy. Whereas gene expression studies by in situ hybridization or immunohistochemistry have shown differential expression patterns of specific genes during implantation, there is no report how numerous signaling proteins are spatially displayed at specific times and stages of implantation in the context of blastocyst-uterine juxtaposition. Using in situ imaging (matrix assisted laser desorption/ionization) mass spectrometry directly on uterine sections, here we provide molecular composition, relative abundance, and spatial distribution of a large number of proteins during the periimplantation period. This approach has allowed us for the first time to generate in situ proteome profiles of implantation and interimplantation sites in mice in a region- and stage-specific manner with the progression of implantation. This application is reliable because patterns of expression of several proteins displayed by in situ imaging mass spectrometry correlate well with in situ hybridization results. More interestingly, the use of this approach has provided new insights regarding uterine biology of cytosolic phospholipase A(2alpha) null females that show implantation defects. PMID- 18403476 TI - Coordinated synchronization in the electrically coupled network of terminal nerve gonadotropin-releasing hormone neurons as demonstrated by double patch-clamp study. AB - The peptidergic neurons play important roles such as neuromodulatory and neuroendocrine functions in the central nervous system. However, our knowledge about the organization and the function of the peptidergic neuromodulator systems is still very poor. The terminal nerve GnRH peptidergic neurons of a teleost, the dwarf gourami (Colisa lalia), serve as an excellent model system for such study. The cell bodies are large and make up a tight cell cluster, and the easy access to the cell bodies on the ventral surface of the brain makes the electrophysiological measurements in a precisely controlled manner. Here we show direct evidence to demonstrate the electrical coupling and the synchronization of the neural firing activity among the terminal nerve GnRH neurons by using the double patch-clamp recording technique. The electrical coupling coefficient was strong enough (ranged from 0.083 to 0.370) to synchronize spontaneous firings of GnRH neurons in the cluster. A model, in which the firings in the cluster occur within a small time window (dozens of milliseconds), was verified by using the serial loose-seal extracellular patch-clamp recordings and the cross-correlogram analysis. The present findings provide several insights for understanding the physiological mechanisms and functional significance of synchronized activities in the peptidergic and/or aminergic neuromodulator system as well as in the peptidergic neuroendocrine cells. PMID- 18403478 TI - Neonatal gonadectomy and adult testosterone replacement suggest an involvement of limbic arginine vasopressin and androgen receptors in the organization of the hypothalamic-pituitary-adrenal axis. AB - Testosterone exposure during critical periods of development exerts major organizing effects on the hypothalamic-pituitary-adrenal (HPA) axis. Here we examined how neonatal gonadectomy (GDX) with or without testosterone treatment during the first week of life alters the HPA response to adult testosterone replacement in 65-d-old male rats. As adults, neonatal GDX rats showed higher levels of plasma corticosterone and Fos activation in medial parvocellular part of the paraventricular nucleus of the hypothalamus under basal conditions and during 30 min of restraint exposure. These responses were normalized with testosterone treatment on postnatal d 1-5 but were not restored with adult testosterone replacement. As adults, neonatal GDX rats also showed a decrease in the number of androgen receptor and arginine vasopressin-positive cells in the bed nucleus of the stria terminalis and in the medial nucleus of the amygdala, and both of these responses were reversed with postnatal testosterone treatment. In stressed and unstressed animals, the number of androgen receptors and arginine vasopressin-expressing neurons in both of these nuclei correlated negatively with corticosterone concentrations in plasma and Fos levels in the paraventricular nucleus. Taken together, our findings suggest that testosterone exposure during the neonatal period primes the adult HPA response to testosterone by altering androgen receptor levels and function within afferent mediators of basal and stress-related input to the HPA axis. PMID- 18403477 TI - Dietary curcumin significantly improves obesity-associated inflammation and diabetes in mouse models of diabesity. AB - Obesity is a major risk factor for the development of type 2 diabetes, and both conditions are now recognized to possess significant inflammatory components underlying their pathophysiologies. We tested the hypothesis that the plant polyphenolic compound curcumin, which is known to exert potent antiinflammatory and antioxidant effects, would ameliorate diabetes and inflammation in murine models of insulin-resistant obesity. We found that dietary curcumin admixture ameliorated diabetes in high-fat diet-induced obese and leptin-deficient ob/ob male C57BL/6J mice as determined by glucose and insulin tolerance testing and hemoglobin A1c percentages. Curcumin treatment also significantly reduced macrophage infiltration of white adipose tissue, increased adipose tissue adiponectin production, and decreased hepatic nuclear factor-kappaB activity, hepatomegaly, and markers of hepatic inflammation. We therefore conclude that orally ingested curcumin reverses many of the inflammatory and metabolic derangements associated with obesity and improves glycemic control in mouse models of type 2 diabetes. This or related compounds warrant further investigation as novel adjunctive therapies for type 2 diabetes in man. PMID- 18403479 TI - Interleukin-6 inhibits receptor activator of nuclear factor kappaB ligand-induced osteoclastogenesis by diverting cells into the macrophage lineage: key role of Serine727 phosphorylation of signal transducer and activator of transcription 3. AB - Osteoclasts are bone-resorptive cells that differentiate from hematopoietic precursors upon receptor activator of nuclear factor kappaB ligand (RANKL) activation. Previous studies demonstrated that IL-6 indirectly stimulates osteoclastogenesis through the production of RANKL by osteoblasts. However, few data described the direct effect of IL-6 on osteoclasts. To investigate this effect, we used several models: murine RAW264.7 cells, mouse bone marrow, and human blood monocytes. In the three models used, the addition of IL-6 inhibited RANKL-induced osteoclastogenesis. Furthermore, IL-6 decreased the expression of osteoclast markers and up-modulated macrophage markers. To elucidate this inhibition, signal transducer and activator of transcription (STAT) 3, the main signaling molecule activated by IL-6, was analyzed. Addition of two STAT3 inhibitors completely abolished RANKL-induced osteoclastogenesis, revealing a key role of STAT3. We demonstrated that a basal level of phosphorylated-STAT3 on Serine(727) associated with an absence of phosphorylation on Tyrosine(705) is essential for osteoclastogenesis. Furthermore, a decrease of Serine(727) phosphorylation led to an inhibition of osteoclast differentiation, whereas an increase of Tyrosine(705) phosphorylation upon IL-6 stimulation led to the formation of macrophages instead of osteoclasts. In conclusion, we showed for the first time that IL-6 inhibits RANKL-induced osteoclastogenesis by diverting cells into the macrophage lineage, and demonstrated the functional role of activated STAT3 and its form of phosphorylation in the control of osteoclastogenesis. PMID- 18403480 TI - Diurnal amplitudes of arterial pressure and heart rate are dampened in Clock mutant mice and adrenalectomized mice. AB - Arterial pressure (AP), heart rate (HR), and cardiovascular diseases, including ischemic heart attack and cerebrovascular accident, show diurnal variation. Evidence that circadian-related genes contribute to cardiovascular control has been accumulated. In this study, we measured the AP and HR of Clock mutant mice on the Jcl/ICR background to determine the role of the Clock gene in cardiovascular function. Mice with mutated Clock genes had a dampened diurnal rhythm of AP and HR, compared with wild-type control mice, and this difference disappeared after adrenalectomy. The diurnal acrophase in both mean arterial pressure and HR was delayed significantly in Clock mutant mice, compared with wild-type mice, and this difference remained after adrenalectomy. Clock mutant mice had a lower concentration of plasma aldosterone, compared with wild-type mice. Our data suggest that the adrenal gland is involved in the diurnal amplitude, but not the acrophase, of AP and HR, and that the function of the Clock gene may be related to the nondipping type of AP elevation. PMID- 18403482 TI - Tumor necrosis factor-alpha attenuates thyroid hormone-induced apoptosis in vascular endothelial cell line XLgoo established from Xenopus tadpole tails. AB - Amphibian metamorphosis induced by T(3) involves programmed cell death and the differentiation of various types of cells in degenerated and reconstructed tissues. However, the signaling pathway that directs the T(3)-dependent cell-fate determinations remains unclear. TNF-alpha is a pleiotropic cytokine that affects diverse cellular responses. Engagement of TNF-alpha with its receptor (TNFR1) causes intracellular apoptotic and/or survival signaling. To investigate TNF signaling functions during anuran metamorphosis, we first identified Xenopus laevis orthologs of TNF (xTNF)-alpha and its receptor. We found that xTNF-alpha activated nuclear factor-kappaB in X. laevis A6 cells through the Fas-associated death domain and receptor-interacting protein 1. Interestingly, xTNF-alpha mRNA in blood cells showed prominent expression at prometamorphosis during metamorphosis. Next, to elucidate the apoptotic and/or survival signaling induced by xTNF-alpha in an in vitro model of metamorphosis, we established a vascular endothelial cell line, XLgoo, from X. laevis tadpole tail. XLgoo cells formed actin stress fibers and elongated in response to xTNF-alpha. T(3) induced apoptosis in these cells, but the addition of xTNF-alpha blocked the T(3)-induced apoptosis. In addition, treatment of the cells with T(3) for 2 d induced the expression of thyroid hormone receptor-beta and caspase-3, and this thyroid hormone receptor-beta induction was drastically repressed by xTNF-alpha. Furthermore, in organ culture of the tail, xTNF-alpha significantly attenuated the tail degeneration induced by T(3). These findings suggested that xTNF-alpha could protect vascular endothelial cells from apoptotic cell death induced by T(3) during metamorphosis and thereby participate in the regulation of cell fate. PMID- 18403481 TI - Corticotropin-releasing hormone deficiency is associated with reduced local inflammation in a mouse model of experimental colitis. AB - CRH, the hypothalamic component of the hypothalamic-pituitary adrenal axis, attenuates inflammation through stimulation of glucocorticoid release, whereas peripherally expressed CRH acts as a proinflammatory mediator. CRH is expressed in the intestine and up-regulated in patients with ulcerative colitis. However, its pathophysiological significance in intestinal inflammatory diseases has just started to emerge. In a mouse model of acute, trinitrobenzene sulfonic acid induced experimental colitis, we demonstrate that, despite low glucocorticoid levels, CRH-deficient mice develop substantially reduced local inflammatory responses. These effects were shown by histological scoring of tissue damage and neutrophil infiltration. At the same time, CRH deficiency was found to be associated with higher serum leptin and IL-6 levels along with sustained anorexia and weight loss, although central CRH has been reported to be a strong appetite suppressor. Taken together, our results support an important proinflammatory role for CRH during mouse experimental colitis and possibly in inflammatory bowel disease in humans. Moreover, the results suggest that CRH is involved in homeostatic pathways that link inflammation and metabolism. PMID- 18403483 TI - Novel expression and direct effects of adiponectin in the rat testis. AB - Adiponectin is an adipocyte hormone, with relevant roles in lipid metabolism and glucose homeostasis, recently involved in the control of different endocrine organs, such as the placenta, pituitary and, likely, the ovary. However, whether as described previously for other adipokines, such as leptin and resistin, adiponectin is expressed and/or conducts biological actions in the male gonad remains unexplored. In this study, we provide compelling evidence for the expression, putative hormonal regulation, and direct effects of adiponectin in the rat testis. Testicular expression of adiponectin was demonstrated along postnatal development, with a distinctive pattern of RNA transcripts and discernible protein levels that appeared mostly located at interstitial Leydig cells. Testicular levels of adiponectin mRNA were marginally regulated by pituitary gonadotropins but overtly modulated by metabolic signals, such as glucocorticoids, thyroxine, and peroxisome proliferator-activated receptor-gamma, whose effects were partially different from those on circulating levels of adiponectin. In addition, expression of the genes encoding adiponectin receptor (AdipoR)-1 and AdipoR2 was detected in the rat testis, with developmental changes and gonadotropin regulation for AdipoR2 mRNA, and prominent levels of AdipoR1 in seminiferous tubules. Moreover, recombinant adiponectin significantly inhibited basal and human choriogonadotropin-stimulated testosterone secretion ex vivo, whereas it failed to change relative levels of several Sertoli cell-expressed mRNAs, such as stem cell factor and anti-Mullerian hormone. In summary, our data are the first to document the expression, regulation and functional role of adiponectin in the rat testis. Taken together with its recently reported expression in the ovary and its effects on LH secretion and ovarian steroidogenesis, these results further substantiate a multifaceted role of adiponectin in the control of the reproductive axis, which might operate as endocrine integrator linking metabolism and gonadal function. PMID- 18403484 TI - Dexamethasone-induced expression of endothelial mitogen-activated protein kinase phosphatase-1 involves activation of the transcription factors activator protein 1 and 3',5'-cyclic adenosine 5'-monophosphate response element-binding protein and the generation of reactive oxygen species. AB - We have recently identified the MAPK phosphatase (MKP)-1 as a novel mediator of the antiinflammatory properties of glucocorticoids (dexamethasone) in the human endothelium. However, nothing is as yet known about the signaling pathways responsible for the up-regulation of MKP-1 by dexamethasone in endothelial cells. Knowledge of the molecular basis of this new alternative way of glucocorticoid action could facilitate the identification of new antiinflammatory drug targets. Thus, the aim of our study was to elucidate the underlying molecular mechanisms. Using Western blot analysis, we found that dexamethasone rapidly activates ERK, c jun N-terminal kinase (JNK), and p38 MAPK in human umbilical vein endothelial cells. By applying the kinase inhibitors PD98059 (MAPK kinase-1) and SP600125 (JNK), ERK and JNK were shown to be crucial for the induction of MKP-1. Using EMSA and a decoy oligonucleotide approach, the transcription factors activator protein-1 (activated by ERK and JNK) and cAMP response element-binding protein (activated by ERK) were found to be involved in the up-regulation of MKP-1 by dexamethasone. Interestingly, dexamethasone induces the generation of reactive oxygen species (measured by dihydrofluorescein assay), which participate in the signaling process by triggering JNK activation. Our work elucidates a novel alternative mechanism for transducing antiinflammatory effects of glucocorticoids in the human endothelium. Thus, our study adds valuable information to the efforts made to find new antiinflammatory principles utilized by glucocorticoids. This might help to gain new therapeutic options to limit glucocorticoid side effects and to overcome resistance. PMID- 18403485 TI - Distinct modes of activation of phosphatidylinositol 3-kinase in response to cyclic adenosine 3', 5'-monophosphate or insulin-like growth factor I play different roles in regulation of cyclin D1 and p27Kip1 in FRTL-5 cells. AB - Bioactivities of IGFs in various cells are often potentiated in the presence of other hormones. In previous studies we showed that pretreatment of rat FRTL-5 thyroid cells with TSH or other cAMP-generating agents markedly potentiated DNA synthesis induced by IGF-I. Under these conditions we found that phosphatidylinositol (PI) 3-kinase was activated in response to either cAMP or IGF stimulus, and both activation modes were indispensable for the potentiation of DNA synthesis. The present studies were undertaken to elucidate how cAMP and/or IGF-I stimulus regulated the G1 cyclin-cyclin dependent kinase (CDK) inhibitor system, and to determine the roles of PI 3-kinase activation by cAMP or IGF-I stimulus in this system. We found that cAMP pretreatment enhanced IGF-I dependent increases in cyclin D1, due to synergistic increases in mRNA and elevation of translation rates. Furthermore, cAMP pretreatment enhanced IGF-I induced protein degradation of the CDK inhibitor, p27(Kip1). These changes well explained an increase in cyclin E, leading to marked activation of G1 CDKs, followed by retinoblastoma protein phosphorylation. Our results using a PI 3 kinase inhibitor showed that cAMP-dependent PI 3-kinase activation plays an important role in the increase in cyclin D1 translation. In contrast, IGF-I dependent PI 3-kinase activation was required for the increase in cyclin D1 mRNA levels and degradation of p27(Kip1). Together, the present study elucidates the role of cAMP and IGF-I in differentially activating PI 3-kinase as a mediator of multiple molecular events. These events converge in the regulation of cyclin D1 and p27(Kip1), leading to cAMP-dependent potentiation of IGF-I-dependent CDK activation and DNA synthesis. PMID- 18403486 TI - Glucocorticoids induce human glycoprotein hormone alpha-subunit gene expression in the gonadotrope. AB - The human glycoprotein hormone alpha-subunit (alphaGSU) gene is transcriptionally regulated by glucocorticoids in a cell type-specific fashion. In direct contrast to repression of alphaGSU by glucocorticoids in placenta, glucocorticoid receptor (GR) modulation in the pituitary is little understood. We show that glucocorticoids stimulate the alphaGSU promoter in immortalized pituitary gonadotrope-derived LbetaT2 cells, whereas estrogens, androgens, and progestins have no significant effect. Moreover, GR acts in a dose-dependent manner at physiological concentrations of glucocorticoids. Transient transfection of GR with dexamethasone (Dex) treatment further stimulates the alphaGSU promoter, but this induction is severely diminished using a receptor mutated in the DNA-binding domain. Truncation and cis mutations demonstrate that glucocorticoid response element 2 (GRE2) and cAMP-response element 2 (CRE2) within -168 bp of the human alphaGSU promoter are critical for induction. Moreover, dominant-negative CRE binding protein markedly inhibits basal but also Dex induction of alphaGSU promoter activity. Additionally, GR specifically binds to GRE2 in the human alphaGSU promoter in vitro and to the 5' region of the endogenous mouse alphaGSU gene in vivo. Furthermore, overexpression of the homeobox factor, Distal-less 3 that regulates this gene in placental cells through a site partially overlapping GRE2, blocks Dex induction of alphaGSU in gonadotrope cells, indicating that placenta-specific expression of Dlx3 may interfere with GR, resulting in repression in placental cells vs. induction in gonadotrope cells. These results demonstrate the stimulatory role played by glucocorticoids in alphaGSU gene expression in the pituitary gonadotrope, in contrast to repression in placental cells, and highlight the tissue-specific nature of steroid hormone action. PMID- 18403487 TI - Signal transducer and activator of transcription-3 is required in hypothalamic agouti-related protein/neuropeptide Y neurons for normal energy homeostasis. AB - Signal transducer and activator of transcription (Stat)-3 signals mediate many of the metabolic effects of the fat cell-derived hormone, leptin. In mice, brain specific depletion of either the long form of the leptin receptor (Lepr) or Stat3 results in comparable obese phenotypes as does replacement of Lepr with an altered leptin receptor locus that codes for a Lepr unable to interact with Stat3. Among the multiple brain regions containing leptin-sensitive Stat3 sites, cells expressing feeding-related neuropeptides in the arcuate nucleus of the hypothalamus have received much of the focus. To determine the contribution to energy homeostasis of Stat3 expressed in agouti-related protein (Agrp)/neuropeptide Y (Npy) arcuate neurons, Stat3 was deleted specifically from these cells, and several metabolic indices were measured. It was found that deletion of Stat3 from Agrp/Npy neurons resulted in modest weight gain that was accounted for by increased adiposity. Agrp/Stat3-deficient mice also showed hyperleptinemia, and high-fat diet-induced hyperinsulinemia. Stat3 deletion in Agrp/Npy neurons also resulted in altered hypothalamic gene expression indicated by increased Npy mRNA and decreased induction of suppressor of cytokine signaling 3 in response to leptin. Agrp mRNA levels in the fed or fasted state were unaffected. Behaviorally, mice without Stat3 in Agrp/Npy neurons were mildly hyperphagic and hyporesponsive to leptin. We conclude that Stat3 in Agrp/Npy neurons is required for normal energy homeostasis, but Stat3 signaling in other brain areas also contributes to the regulation of energy homeostasis. PMID- 18403488 TI - Dendritic action potential initiation in hypothalamic gonadotropin-releasing hormone neurons. AB - It is dogma that action potentials are initiated at the soma/axon hillock of neurons. However, dendrites often exhibit conductances necessary for spike generation and represent functionally independent processing compartments within neurons. GnRH neurons provide an interesting neuronal phenotype with simple, relatively unbranched, unipolar or bipolar dendrites of extensive lengths (>1000 microm) covered in spines. These neurons control fertility and must integrate a variety of internal homeostatic and external environmental cues. We used imaging, electrophysiological, and modeling studies to understand how they integrate and process information along dendrites. Simultaneous recordings from distal dendrites and somata of individual GnRH neurons indicate distal dendrites are the primary site of spike initiation in these cells. Compartmental modeling indicates that sites of spike initiation depend upon location of excitatory input and dendrite geometry. Together, these studies demonstrate a novel pattern of spike generation in mammalian neurons and indicate that afferent inputs within distal dendritic microdomains directly initiate action potentials. PMID- 18403489 TI - Spermatogenesis and sertoli cell activity in mice lacking sertoli cell receptors for follicle-stimulating hormone and androgen. AB - Spermatogenesis in the adult male depends on the action of FSH and androgen. Ablation of either hormone has deleterious effects on Sertoli cell function and the progression of germ cells through spermatogenesis. In this study we generated mice lacking both FSH receptors (FSHRKO) and androgen receptors on the Sertoli cell (SCARKO) to examine how FSH and androgen combine to regulate Sertoli cell function and spermatogenesis. Sertoli cell number in FSHRKO-SCARKO mice was reduced by about 50% but was not significantly different from FSHRKO mice. In contrast, total germ cell number in FSHRKO-SCARKO mice was reduced to 2% of control mice (and 20% of SCARKO mice) due to a failure to progress beyond early meiosis. Measurement of Sertoli cell-specific transcript levels showed that about a third were independent of hormonal action on the Sertoli cell, whereas others were predominantly androgen dependent or showed redundant control by FSH and androgen. Results show that FSH and androgen act through redundant, additive, and synergistic regulation of spermatogenesis and Sertoli cell activity. In addition, the Sertoli cell retains a significant capacity for activity, which is independent of direct hormonal regulation. PMID- 18403490 TI - Starvation and triglycerides reverse the obesity-induced impairment of insulin transport at the blood-brain barrier. AB - Insulin in the brain acts as a satiety factor, reduces appetite, and decreases body mass. Altered sensing by brain of insulin may be a leading cause of weight gain and insulin resistance. A decrease in the transport across the blood-brain barrier (BBB) of insulin may induce brain insulin resistance by inducing obesity. We here report that transport of iv administrated insulin across the BBB of obese mice, as measured by multiple-time regression analysis, was significantly lower than that in thin adult mice. The reduction in obese mice was reversed by starvation for 48 h. There were no differences in insulin transport rates across the BBB of obese, thin, or starved obese mice when studied by the brain perfusion model, demonstrating that BBB transport of insulin is modulated by circulating factors. In the brain perfusion study, the triglyceride triolein significantly increased the brain uptake of insulin, an effect opposite to that on leptin transport, in starved obese mice. Thus, circulating triglycerides are one of the systemic modulators for the transport of insulin across the BBB. PMID- 18403491 TI - Intracerebral infusion of an EGFR-targeted toxin in recurrent malignant brain tumors. AB - The purpose of this study is to determine the maximum tolerated dose (MTD), dose limiting toxicity (DLT), and intracerebral distribution of a recombinant toxin (TP-38) targeting the epidermal growth factor receptor in patients with recurrent malignant brain tumors using the intracerebral infusion technique of convection enhanced delivery (CED). Twenty patients were enrolled and stratified for dose escalation by the presence of residual tumor from 25 to 100 ng/ml in a 40-ml infusion volume. In the last eight patients, coinfusion of (123)I-albumin was performed to monitor distribution within the brain. The MTD was not reached in this study. Dose escalation was stopped at 100 ng/ml due to inconsistent drug delivery as evidenced by imaging the coinfused (123)I-albumin. Two DLTs were seen, and both were neurologic. Median survival after TP-38 was 28 weeks (95% confidence interval, 26.5-102.8). Of 15 patients treated with residual disease, two (13.3%) demonstrated radiographic responses, including one patient with glioblastoma multiforme who had a nearly complete response and remains alive >260 weeks after therapy. Coinfusion of (123)I-albumin demonstrated that high concentrations of the infusate could be delivered >4 cm from the catheter tip. However, only 3 of 16 (19%) catheters produced intraparenchymal infusate distribution, while the majority leaked infusate into the cerebrospinal fluid spaces. Intracerebral CED of TP-38 was well tolerated and produced some durable radiographic responses at doses first quartile 2.245, 95% CI 1.371 to 3.677). Severity of oesophagitis was only associated with acid exposure (OR up to 5.038, 95% CI 1.452 to 17.480). The presence of Barrett's oesophagus was associated with male sex (OR 3.621, 95% CI 2.263 to 5.794), DGOR (OR up to 5.017, 95% CI 2.051 to 12.274) and acid exposure (OR up to 3.031, 95% CI 1.216 to 7.556). CONCLUSIONS: Several independent factors are associated with oesophageal lesions in GORD. The risk of oesophagitis is associated with hiatal hernia, BMI and DGOR exposure; severity of oesophagitis depends on acid exposure; and Barrett's oesophagus is associated with male sex and exposure to both acid and DGOR. PMID- 18403497 TI - Assessment of proteinuria in pregnancy. PMID- 18403498 TI - Diagnostic accuracy of urinary spot protein:creatinine ratio for proteinuria in hypertensive pregnant women: systematic review. AB - OBJECTIVE: To review the spot protein:creatinine ratio and albumin:creatinine ratio as diagnostic tests for significant proteinuria in hypertensive pregnant women. DESIGN: Systematic review. DATA SOURCES: Medline and Embase, the Cochrane Library, reference lists, and experts. Review methods Literature search (1980 2007) for articles of the spot protein:creatinine ratio or albumin:creatinine ratio in hypertensive pregnancy, with 24 hour proteinuria as the comparator. RESULTS: 13 studies concerned the spot protein:creatinine ratio (1214 women with primarily gestational hypertension). Nine studies reported sensitivity and specificity for eight cut-off points, median 24 mg/mmol (range 17-57 mg/mmol; 0.15-0.50 mg/mg). Laboratory assays were not well described. Diagnostic test characteristics were recalculated for a cut-off point of 30 mg/mmol. No significant heterogeneity in cut-off points was found between studies over a range of proteinuria. Pooled values gave a sensitivity of 83.6% (95% confidence interval 77.5% to 89.7%), specificity of 76.3% (72.6% to 80.0%), positive likelihood ratio of 3.53 (2.83 to 4.49), and negative likelihood ratio of 0.21 (0.13 to 0.31) (nine studies, 1003 women). Two studies of the spot albumin:creatinine ratio (225 women) found optimal cut-off points of 2 mg/mmol for proteinuria of 0.3 g/day or more and 27 mg/mmol for albuminuria. CONCLUSION: The spot protein:creatinine ratio is a reasonable "rule-out" test for detecting proteinuria of 0.3 g/day or more in hypertensive pregnancy. Information on use of the albumin:creatinine ratio in these women is insufficient. PMID- 18403499 TI - Migration and health behaviour during pregnancy. PMID- 18403501 TI - Selecting medical students. PMID- 18403502 TI - Direct to consumer advertising: A cynical consultation exercise? PMID- 18403500 TI - Influence of moving to the UK on maternal health behaviours: prospective cohort study. AB - OBJECTIVE: To compare health behaviours during pregnancy (smoking and alcohol consumption) and after birth (initiation and duration of breast feeding) between British/Irish white mothers and mothers from ethnic minority groups; and, in mothers from ethnic minority groups, to examine whether indicators of acculturation (generational status, language spoken at home, length of residency in the United Kingdom) were associated with these health behaviours. DESIGN: Prospective nationally representative cohort study. SETTING: England. PARTICIPANTS: 6478 British/Irish white mothers and 2110 mothers from ethnic minority groups. MAIN OUTCOME MEASURES: Any smoking during pregnancy; any alcohol consumption during pregnancy; initiation of breast feeding; breast feeding for at least four months. RESULTS: Compared with British/Irish white mothers, mothers from ethnic minority groups were less likely to smoke (15% v 37%) or consume alcohol (14% v 37%) during pregnancy but more likely to initiate breast feeding (86% v 69%) and breast feed for at least four months (40% v 27%). Among mothers from ethnic minority groups, first and second generation mothers were more likely to smoke during pregnancy (odds ratio 3.85, 95% confidence interval 2.50 to 5.93, and 4.70, 2.49 to 8.90, respectively), less likely to initiate breast feeding (0.92, 0.88 to 0.97, and 0.86, 0.75 to 0.99), and less likely to breast feed for at least four months (0.72, 0.62 to 0.83, and 0.52, 0.30 to 0.89) than immigrants, after adjustment for sociodemographic characteristics. There were no consistent differences in alcohol consumption. Among immigrants, for every additional five years spent in the UK the likelihood of mothers smoking during pregnancy increased by 31% (4% to 66%) and they were 5% (0% to 10%) less likely to breast feed for at least four months. CONCLUSIONS: After immigration, maternal health behaviours worsen with length of residency in the UK. Health professionals should not underestimate women's likelihood of engaging in risky health behaviours because of their ethnicity. PMID- 18403503 TI - Over the counter medicines: Don't include trimethoprim. PMID- 18403504 TI - Health of the workers: Another ethical muddle for GMC. PMID- 18403505 TI - Health of the workers: Sick sick notes (sic). PMID- 18403506 TI - UKCAT among the pigeons... and excluding the empathic. PMID- 18403507 TI - UKCAT among the pigeons: UKCAT among the pigeons. Undermining access ... PMID- 18403508 TI - Direct to consumer advertising: Include cost of treatment. PMID- 18403509 TI - Ditching impact factors: Time for the single researcher impact factor. PMID- 18403510 TI - Predicting citations: Validating prediction models. PMID- 18403511 TI - Health of the workers: Sick note, fit note, no note? PMID- 18403512 TI - Computer says yes: Let the patient decide. PMID- 18403513 TI - Multiple health problems: Presentation is important too. PMID- 18403514 TI - GMC guidance on beliefs: Denies conscientious objection. PMID- 18403515 TI - Robotic prostatectomy: Data, please. PMID- 18403516 TI - Gynaecomastia: Are surgical guidelines realistic? PMID- 18403517 TI - Gynaecomastia: Drugs and surgical concerns. PMID- 18403518 TI - BMA angered by below inflation pay offer. PMID- 18403519 TI - Paediatricians debate a motion of no confidence in General Medical Council. PMID- 18403524 TI - Canadian Medical Association launches online health consultations. PMID- 18403520 TI - EC proposals fail to distinguish between information and advertising. PMID- 18403526 TI - Watchdog says trusts must improve their handling of complaints. PMID- 18403529 TI - Unicef reports progress in cutting vertical transmission of HIV. PMID- 18403532 TI - UK doctors show most interest in falls among elderly people, but French more curious about type 2 diabetes. PMID- 18403537 TI - Plagiarism detection service to be launched in June. PMID- 18403538 TI - US ready for healthcare reform, conference told. PMID- 18403540 TI - Donors agree financial arrangement to stimulate vaccine development. PMID- 18403541 TI - Suicide and the internet. PMID- 18403542 TI - Safety nets. PMID- 18403543 TI - Does the NHS really need a constitution? PMID- 18403544 TI - Competition and integration in the English National Health Service. PMID- 18403545 TI - Let patients control the purse strings. PMID- 18403546 TI - Catheter ablation for atrial fibrillation. PMID- 18403547 TI - The self critical doctor: helping students become more reflective. PMID- 18403548 TI - Pulmonary venous stenosis after treatment for atrial fibrillation. PMID- 18403549 TI - Suspected premature menopause. PMID- 18403550 TI - Radical Muslim doctors and what they mean for the NHS. PMID- 18403554 TI - MR imaging criteria for the prediction of extranodal spread of metastatic cancer in the neck. AB - BACKGROUND AND PURPOSE: The presence of extranodal spread in metastatic nodes significantly affects treatment planning and prognosis of the patient with head and neck cancer. We attempted to evaluate the predictive capability of MR imaging for the extranodal spread in the neck. MATERIALS AND METHODS: We retrospectively studied MR images from 109 patients with histologically proved metastatic nodes, of which 39 were positive for extranodal spread. We assessed 47 extranodal spread positive and 130 extranodal spread-negative metastatic nodes by using the following MR imaging findings as the possible criteria for extranodal spread: 1) nodal size (short-axis diameter); 2) obliterated fat spaces between the metastatic node and adjacent tissues, such as the muscles and skin on T1-weighted images ("vanishing border" sign); 3) the presence of high-intensity signals in the interstitial tissues around and extending from a metastatic node on fat suppressed T2-weighted images ("flare" sign); and 4) an irregular nodal margin on gadolinium-enhanced T1-weighted images ("shaggy margin"). Multivariate logistic regression analysis was conducted to identify independent predictive criteria for extranodal spread. RESULTS: Nodal size, shaggy margin, and flare sign criteria were independent and significant MR imaging findings suggestive of extranodal spread in the metastatic nodes. We obtained 77% sensitivity and 93% specificity with the flare sign, 65% sensitivity and 99% specificity with the shaggy margin, and 80% sensitivity and 85% specificity with the size criterion (cutoff point = 16 mm). CONCLUSION: Fat-suppressed T2-weighted and gadolinium-enhanced T1 weighted images are useful for the detection of extranodal spread in metastatic nodes in the neck. PMID- 18403556 TI - Local intra-arterial eptifibatide for intraoperative vessel thrombosis during aneurysm coiling. AB - We report on our experience with the intra-arterial administration of eptifibatide for thrombolysis during aneurysm-embolization procedures. In 4 cases (3 stent-assisted coiling procedures and 1 with posthemorrhagic vasospasm), we noted the formation of thrombus occluding a vessel. We administered eptifibatide (10-15 mg) through a microcatheter proximal to the thrombus. The thrombus rapidly dissolved, resulting in the recanalization of the occluded vessels with no rethrombosis or hemorrhagic complications. PMID- 18403555 TI - Multiphase CT angiography versus single-phase CT angiography: comparison of image quality and radiation dose. AB - BACKGROUND AND PURPOSE: Conventional CT angiography (CTA) is acquired during only a short interval in the arterial phase, which limits its ability to evaluate the cerebral circulation. Our aim was to compare the image quality and radiation dose of conventional single-phase CTA (SP-CTA) with a multiphase CTA (MP-CTA) algorithm reconstructed from a perfusion CT (PCT) dataset. MATERIALS AND METHODS: Fifty consecutive patients undergoing head CTA and PCT in 1 examination were enrolled. The PCT dataset was obtained with 40.0-mm-detector coverage, 5.0-mm axial thickness, 80 kilovolt peak (kVp), 180 mA, and 30 mL of contrast medium. MP CTA was reconstructed from the same PCT dataset with an axial thickness of 0.625 mm by using a new axial reconstruction algorithm. A conventional SP-CTA dataset was obtained with 0.625-mm axial thickness, 120 kVp, 350 mA, and 60 mL of contrast medium. We compared image quality, vascular enhancement, and radiation dose. RESULTS: SP-CTA and MP-CTA of 50 patients (male/female ratio, 31/19; mean age, 59.25 years) were analyzed. MP-CTA was significantly better than SP-CTA in vascular enhancement (P = .002), in the absence of venous contamination (P = .006), and was significantly higher in image noise (P < .001). MP-CTA used less contrast medium than SP-CTA and could demonstrate hemodynamic information. The effective dose of MP-CTA was 5.73 mSv, which was equal to that in conventional PCT, and it was 3.57 mSv in SP-CTA. CONCLUSION: It is feasible that MP-CTA may provide both CTA and PCT results. Compared with SP-CTA, MP-CTA provides comparable image quality, better vascular enhancement, hemodynamic information, and more noise with less detail visibility with a lower tube voltage. The radiation dose of MP-CTA is higher than that of SP-CTA, but the dose can be reduced by altering the sampling interval. PMID- 18403557 TI - Immediate shrinkage of optociliary shunt vessels after fractionated external beam radiation for meningioma of the optic nerve sheath. AB - Fractionated stereotactic radiation has become the standard treatment of meningioma of the optic nerve sheath. The mechanism responsible for improvement in visual function is unclear, because neuroimaging after treatment usually shows no discernable change in tumor appearance. We report immediate regression of optociliary shunt vessels in a patient after radiation treatment of an optic nerve sheath meningioma. This observation indicates that radiation treatment can cause rapid reduction of optic nerve compression, even without appreciable reduction in the size of the meningioma. PMID- 18403558 TI - Sinonasal organized hematoma: CT and MR imaging findings. AB - BACKGROUND AND PURPOSE: Sinonasal organized hematoma (OH) is an uncommon, nonneoplastic benign condition that can be locally aggressive. The purpose of this work was to characterize the CT and MR imaging findings of sinonasal OH. MATERIALS AND METHODS: CT (n = 11) and MR (n = 10) images of 12 patients (9 men and 3 women; mean age, 41 years; range, 12-76 years) with pathologically proved sinonasal OH were retrospectively reviewed. Particular attention was put on the location, shape, size, extent, internal architecture, and enhancement pattern of the lesion and associated sinus wall change. RESULTS: The lesions were seen as an expansile (n = 9) or nonexpansile (n = 3) mass, ranging in size from 2.2 to 6.0 cm (mean, 4.2 cm), primarily involving the maxillary sinus (n = 11) or nasal cavity (n = 1) unilaterally. The ipsilateral nasal cavity was also involved in 9 of 11 maxillary sinus lesions. Smooth sinus wall erosion other than the medial maxillary sinus wall was noted in 8 lesions. The internal architecture was best displayed on T2-weighted MR images on which all of the lesions were seen as a mixture of marked heterogeneous hypointensity and isointensity, surrounded by a hypointense peripheral rim, reflecting histologic heterogeneity of the lesion composed of hemorrhage, fibrosis, and neovascularization. Marked irregular nodular, papillary, or frondlike enhancement at the areas of neovascularization was also a typical finding seen in all of the lesions. CONCLUSION: An expansile soft tissue mass, smooth sinus wall erosion, marked heterogeneous signal intensity with a hypointense peripheral rim on T2-weighted MR images, and marked irregular nodular, papillary, or frondlike enhancement are characteristic CT and MR imaging findings of sinonasal OH. PMID- 18403559 TI - CT perfusion identifies increased salvage of tissue in patients receiving intravenous recombinant tissue plasminogen activator within 3 hours of stroke onset. AB - BACKGROUND AND PURPOSE: In spite of the advent of thrombolytic therapy, CT perfusion imaging is currently not fully used for clinical decision-making and not included in published clinical guidelines for management of ischemic stroke. We investigated whether lesion volumes on cerebral blood volume (CBV), cerebral blood flow (CBF), and mean transit time (MTT) maps predict final infarct volume and whether all these parameters are needed for triage to intravenous recombinant tissue plasminogen activator (rtPA). We also investigated the effect of intravenous rtPA on affected brain by measuring salvaged tissue volume in patients receiving intravenous rtPA and in controls. MATERIALS AND METHODS: Forty four patients receiving intravenous rtPA and 19 controls underwent CT perfusion (CTP) studies in the emergency department within 3 hours of stroke onset. Lesion volumes were measured on MTT, CBV, and CBF maps by region-of-interest analysis and were compared with follow-up CT volumes by correlation and regression analysis. The volume of salvaged tissue was determined as the difference between the initial MTT and follow-up CT lesion volumes and was compared between intravenous rtPA-treated patients and controls. RESULTS: No significant difference between the groups was observed in lesion volume assessed from the CTP maps (P > .08). Coefficients of determination for MTT, CBF, and CBV versus follow up CT lesion volumes were 0.3, 0.3, 0.47, with intravenous rtPA; and 0.53, 0.55, and 0.81 without intravenous rtPA. Regression of MTT on CBF lesion volumes showed codependence (R(2) = 0.98, P < .0001). Mean salvaged tissue volumes with intravenous rtPA were 21.8 +/- 17.1 and 13.2 +/- 13.5 mL in controls; these were significantly different by using nonparametric (P < .03) and Fisher exact tests (P < .04). CONCLUSIONS: Within 3 hours of stroke onset, CBV lesion volume does not necessarily represent dead tissue. MTT lesion volume alone can be used to identify the upper limit of the size of abnormally perfused brain. More brain is salvaged in patients with intravenous rtPA than in controls. PMID- 18403560 TI - Posterior reversible encephalopathy syndrome, part 2: controversies surrounding pathophysiology of vasogenic edema. AB - Posterior reversible encephalopathy syndrome (PRES) is a neurotoxic state accompanied by a unique brain imaging pattern typically associated with a number of complex clinical conditions including: preeclampsia/eclampsia, allogeneic bone marrow transplantation, solid organ transplantation, autoimmune diseases and high dose cancer chemotherapy. The mechanism behind the developing vasogenic edema and CT or MR imaging appearance of PRES is not known. Two theories have historically been proposed: 1) Severe hypertension leads to failed auto-regulation, subsequent hyperperfusion, with endothelial injury/vasogenic edema and; 2) vasoconstriction and hypoperfusion leads to brain ischemia and subsequent vasogenic edema. The strengths/weaknesses of these hypotheses are reviewed in a translational fashion including supporting evidence and current available imaging/clinical data related to the conditions that develop PRES. While the hypertension/hyperperfusion theory has been most popular, the conditions associated with PRES have a similar immune challenge present and develop a similar state of T-cell/endothelial cell activation that may be the basis of leukocyte trafficking and systemic/cerebral vasoconstriction. These systemic features along with current vascular and perfusion imaging features in PRES appear to render strong support for the older theory of vasoconstriction coupled with hypoperfusion as the mechanism. PMID- 18403561 TI - Modified HPLC-electrospray ionization/mass spectrometry method for HbA1c based on IFCC reference measurement procedure. AB - BACKGROUND: Monitoring of hemoglobin A(1c) (HbA(1c)) is important in the management of diabetes. The IFCC reference measurement procedure for HbA(1c) is based on the ratio of glycated to nonglycated N-terminal hexapeptides of the beta chains of hemoglobin after digestion with Glu-C endoproteinase. We developed a modification of the original reference measurement procedure with HPLC electrospray ionization/mass spectrometry (ESI/MS). METHOD: We performed chromatographic separation of the hexapeptides using a C12 reversed-phase column and a binary gradient system consisting of a mixture of H(2)O/acetonitrile/formic acid. RESULTS: Using this method, we obtained higher signal intensities and improved system stability compared with the reference measurement procedure. In the range of 3% to 14% HbA(1c), intralaboratory CVs were 0.71% to 1.86%. Deviations from IFCC target values were -0.87 to 1.00 relative %. These values fulfill acceptability criteria for HbA(1c) determination set by the IFCC Working Group on HbA(1c) Standardization. CONCLUSIONS: This procedure for the determination of HbA(1c) improves the existing reference measurement procedure. PMID- 18403562 TI - Design and application of noncontinuously binding probes used for haplotyping and genotyping. AB - BACKGROUND: Many methods for genotyping use melting temperature (Tm) of sequence specific probes. Usually the probes hybridize to a continuous stretch of DNA that contains the variant(s). In contrast, hybridization of noncontinuous probes to a template can form bulges. This report generates guidelines for the design of noncontinuous probes. METHODS: We used software to predict hybridization structures and Tms from 10 noncontinuous probes and 54 different templates. Predicted Tms were compared to existing experimental data. The bulging template's sequences (omitted in the probe) ranged in size from 1 to 73 nucleotides. In 36 cases, we compared observed and predicted DeltaTms between alleles complementary to the probe and mismatched alleles. In addition, using software that predicts effects of bulges, we designed a probe and then tested it experimentally. RESULTS: The mean differences between predicted and observed Tms were 0.65 (2.51) degrees C with the Visual OMP software and 0.28 (1.67) degrees C with the MeltCalc software. DeltaTms were within a mean (SD) of 0.36 (1.23) degrees C (Visual OMP) and -0.01 (1.02) degrees C (MeltCalc) of observed values. An increase in the size of the template bulge resulted in a decrease in Tms. In 2 templates, the presence of a variant in the bulge influenced the experimental Tm of 2 noncontinuous probes, a result that was not predicted by the software programs. CONCLUSIONS: The use of software prediction should prove useful for the design of noncontinuous probes that can be used as tools for molecular haplotyping, multiplex genotyping, or masking sequence variants. PMID- 18403563 TI - Ethnic differences in C-reactive protein concentrations. AB - BACKGROUND: Limited data exist regarding the ethnic differences in C-reactive protein (CRP) concentrations, an inflammatory marker associated with risk of cardiovascular disease (CVD). We hypothesized that known CVD risk factors, including anthropometric characteristics, would explain much of the observed ethnic variation in CRP. METHODS: We performed a cross-sectional analysis of 3154 women, without known CVD and not receiving hormone therapy, enrolled in the Study of Women's Health Across the Nation (SWAN), a multiethnic prospective study of pre- and perimenopausal women. RESULTS: The study population was 47.4% white, 27.7% African-American, 8.5% Hispanic, 7.7% Chinese, and 8.6% Japanese; mean age was 46.2 years. African-American women had the highest median CRP concentrations (3.2 mg/L), followed by Hispanic (2.3 mg/L), white (1.5 mg/L), Chinese (0.7 mg/L), and Japanese (0.5 mg/L) women (all pairwise P < 0.001 compared with white women). Body mass index (BMI) markedly attenuated the association between ethnicity and CRP. After adjusting for age, socioeconomic status, BMI, and other risk factors, African-American ethnicity was associated with CRP concentrations >3 mg/L (odds ratio 1.37, 95% CI 1.07-1.75), whereas Chinese and Japanese ethnicities were inversely related (0.58, 0.35-0.95, and 0.43, 0.26-0.72, respectively). CONCLUSIONS: Modifiable risk factors, particularly BMI, account for much but not all of the ethnic differences in CRP concentrations. Further study is needed of these ethnic differences and their implications for the use of CRP in CVD risk prediction. PMID- 18403564 TI - Rapid and sensitive detection of BRCA1/2 mutations in a diagnostic setting: comparison of two high-resolution melting platforms. AB - BACKGROUND: High-resolution melting is an emerging technique for detection of nucleic acid sequence variations. Developments in instrumentation and saturating intercalating dyes have made accurate high-resolution melting analysis possible and created opportunities to use this technology in diagnostic settings. We evaluated 2 high-resolution melting instruments for screening BRCA1 and BRCA2 mutations. METHODS: To cover the complete coding region and splice sites, we designed 112 PCR amplicons (136-435 bp), amplifiable with a single PCR program. LCGreen Plus was used as the intercalating dye. High-resolution melting analysis was performed on the 96-well Lightscanner (Idaho Technology Inc.) and the 96-well LightCycler 480 (Roche) instruments. We evaluated sensitivity by analyzing 212 positive controls scattered over almost all amplicons and specificity by blind screening of 22 patients for BRCA1 and BRCA2. In total, we scanned 3521 fragments. RESULTS: All 212 known heterozygous sequence variants were detected on the Lightscanner by analysis on normal sensitivity setting. On the LightCycler 480, the standard instrument sensitivity setting of 0.3 had to be increased to 0.7 to detect all variants, decreasing the specificity to 95.9% (vs 98.7% for the Lightscanner). CONCLUSIONS: Previously, we screened BRCA1/2 by direct sequencing of the large exon 11 and denaturing gel gradient electrophoresis (DGGE) for all other coding exons. Since the introduction of high-resolution melting, our turnaround time has been one third of that with direct sequencing and DGGE, as post-PCR handling is no longer required and the software allows fast analyses. High-resolution melting is a rapid, cost-efficient, sensitive method simple enough to be readily implemented in a diagnostic laboratory. PMID- 18403565 TI - Log-PCR: a new tool for immediate and cost-effective diagnosis of up to 85% of dystrophin gene mutations. AB - BACKGROUND: Duchenne (DMD) and Becker (BMD) muscular dystrophies are caused by mutations in the dystrophin gene. Despite the progress in the technologies of mutation detection, the disease of one third of patients escapes molecular definition because the labor and expense involved has precluded analyzing the entire gene. Novel techniques with higher detection rates, such as multiplex ligation-dependent probe amplification and multiplex amplifiable probe hybridization, have been introduced. METHODS: We approached the challenge of multiplexing by modifying the PCR chemistry. We set up a rapid protocol that analyzes all dystrophin exons and flanking introns (57.5 kb). We grouped exons according to their effect on the reading frame and ran 2 PCR reactions for DMD mutations and 2 reactions for BMD mutations under the same conditions. The PCR products are evenly spaced logarithmically on the gel (Log-PCR) in an order that reproduces their chromosomal locations. This strategy enables both simultaneous mapping of all the mutation borders and distinguishing between DMD and BMD. As a proof of principle, we reexamined samples from 506 patients who had received a DMD or BMD diagnosis. RESULTS: We observed gross rearrangements in 428 of the patients (84.6%; 74.5% deletions and 10.1% duplications). We also recognized a much broader spectrum of mutations and identified 14.6% additional cases. CONCLUSIONS: This study is the first exhaustive investigation of this subject and has made possible the development of a cost-effective test for diagnosing a larger proportion of cases. The benefit of this approach may allow more focused efforts for discovering small or deep-intronic mutations among the few remaining undiagnosed cases. The same protocol can be extended to set up Log-PCRs for other high-throughput applications. PMID- 18403566 TI - Affinity maturation of tacrolimus antibody for improved immunoassay performance. AB - BACKGROUND: Organic solvents used for extraction of tacrolimus from whole blood samples lower the apparent affinity of the antibody used in a diagnostic immunoassay, thereby affecting the detection limit. METHODS: We used in vitro recombinant antibody engineering to screen and isolate clones from diverse libraries with mutagenic complementarity regions (CDRs) from tacrolimus 1-60-46 hybridoma cell line, with improved binding to tacrolimus in the presence of 10% methanol organic solvent solution. RESULTS: We isolated a number of clones with mutations in variable heavy (VH) CDR 2, variable light (VL) CDR 1, and VL CDR 3 with improved binding. Various combinatorial pairings constructed from these individual mutations contained >10-fold improvements in both the dissociation rate and overall equilibrium affinity constants. Selected clones produced as IgG have increased functional sensitivity, with a 3- to 6-fold reduction in the limit of detection relative to the parental tacrolimus 1-60-46 monoclonal antibody in the Architect Tacrolimus immunodiagnostic assay. CONCLUSIONS: The recent advent of recombinant in vitro antibody display technologies in general, and yeast surface display in particular, allows the flexibility to engineer new or augment specific analytical characteristics, such as affinity, specificity, or stability, into previously isolated and otherwise desirable antibodies to enhance assay performance. These in vitro selections can also be performed under conditions meant to mimic the assay in which the reagent will ultimately be used, to increase the likelihood of successful assay development. PMID- 18403567 TI - Single-step scalable-throughput molecular screening for Huntington disease. AB - BACKGROUND: Huntington disease (HD) is a fatal autosomal dominant neurodegenerative disorder caused by an unstable expansion of the CAG trinucleotide repeat in exon 1 of the HTT (huntingtin) gene and typically has an adult onset. Molecular diagnosis and screening for HD currently involve separate amplification and detection steps. METHODS: We evaluated a novel, rapid microplate-based screening method for HD that combines the amplification and detection procedures in a single-step, closed-tube format. We carried out both the PCR for the HTT CAG-repeat region and the subsequent automated melting-curve analysis of the amplicon in the same wells on the plate. To establish cutoff melting temperatures (T(m)s) for each allelic class, we used a panel of reference DNA samples of known CAG-repeat sizes that represent a range of HTT alleles [normal (< or =26 repeats), intermediate (27-35 repeats), reduced penetrance expanded (36-39 repeats), and fully penetrant expanded (> or =40 repeats)]. We also measured well-to-well variation in T(m) across the thermal block and validated cutoff T(m)s with DNA samples from 5 different populations. We also conducted a blinded validation analysis of clinical samples from an additional 40 HD-affected and 30 unaffected individuals. RESULTS: We observed a strong correlation between CAG-repeat size and amplicon T(m) among the reference DNA samples. Use of the T(m) cutoffs we established revealed that 5 samples from unaffected individuals had been misclassified as affected (1.1% false-positive rate). All samples from HD-affected and unaffected individuals were correctly identified in the blinded analysis. CONCLUSIONS: This simple and scalable homogeneous assay may serve as a convenient, rapid, and accurate screen to detect the presence of pathologic expanded HD alleles in symptomatic patients. PMID- 18403568 TI - After ENHANCE: is more LDL cholesterol lowering even better? PMID- 18403569 TI - Arabic translation and adaptation of Critical Care Family Satisfaction Survey. AB - OBJECTIVE: To translate and adapt the Critical Care Family Satisfaction Survey (CCFSS), and test its validity and reliability for use in Saudi Arabia. SETTING: Seven hundred-bed tertiary care hospital in Saudi Arabia. PARTICIPANTS: Seventy six adult relatives of patients who had been cared for in an intensive care unit for 24 hrs or more. INTERVENTIONS: The CCFSS, a battery of 20 items divided into five subscales, was translated into Arabic. After transfer of patients to regular inpatient units, interviewers administered the survey to their next-of-kin. Respondents ranked their satisfaction with each item on a 5-point Likert scale. MAIN OUTCOME MEASURES: Subscale scores were calculated as the average of the individual item scores. The total scale score was the sum of the subscale scores. RESULTS: The total scale and 'Information', 'Support', 'Comfort' and 'Assurance' subscales showed acceptable internal consistency (Spearman's correlation coefficient of the total score with each of the subscale scores = 0.52-0.81, P<0.01; Cronbach's alpha = 0.67-0.88). But the 'Proximity' subscale performed poorly (r=0.48, P<0.01; Cronbach's alpha = 0.36). Discriminant validity was tested with a Spearman's rank correlation matrix of the subscales, and ranged from weak between 'Support' and both 'Assurance' and 'Information' (r=0.80) to substantial between 'Information' and 'Proximity' (r=0.54) (P<0.01). CONCLUSION: This Arabic translation and adaptation of the CCFSS is a valid, reliable and feasible tool to evaluate family satisfaction in Saudi Arabian intensive care units. PMID- 18403570 TI - Developing clinical indicators for the secondary health system in India. AB - QUALITY PROBLEM OR ISSUE: One of the prime goals of any health system is to deliver good and competent quality of healthcare. Through World Bank-assisted Maharashtra Health Systems Development Project, Government of Maharashtra in India developed and implemented clinical indicators to improve quality. INITIAL ASSESSMENT: During this, clinical areas eligible for monitoring quality of care and roles of health staff working at various levels were identified. CHOICE OF SOLUTION: Brainstorming discussion sessions were conducted to refine list of potential clinical indicators and to identify implementation problems. IMPLEMENTATION: It was implemented in four stages. (a) Self-explanatory tool of record, standard operating procedures and training manual were prepared during tools preparation stage. (b) Pilot implementation was done to monitor the usefulness of indicators, document the experiences and standardize the system accordingly. (c) The final selection of indicators was done taking into consideration points like data reliability, indicator usefulness etc. For final implementation, 15 indicators for district and 6 indicators for rural hospitals were selected. (d) Transfer of skills was done through training of various hospital functionaries. EVALUATION AND LESSONS LEARNED: Selection and prioritization of clinical indicators is the most crucial part. Active participation of local employees is essential for sustainability of the scheme. It is also important to ensure that data recorded/reported is both reliable and valid, to conduct monthly review of the scheme at various levels and to link it with the quality improvement programme. PMID- 18403571 TI - Personality and all-cause mortality among older adults dwelling in a Japanese community: a five-year population-based prospective cohort study. AB - OBJECTIVE: Personality is an important factor in determining longevity. It has been reported that some personality traits can affect mortality via health related behaviors, engaging in social interactions, and a tendency to experience negative emotions. The authors examined the relationships between five major domains of personality traits and all-cause mortality among Japanese community dwelling elderly. DESIGN: A 5-year prospective cohort study design with mortality surveillance. SETTING: Community based. PARTICIPANTS: A total sample of 486 men and 742 women aged 65 years and over at the baseline. MEASUREMENTS: The NEO five factor inventory was administered to assess the "big five" personality traits: neuroticism, extraversion, openness, agreeableness, and conscientiousness. RESULTS: During the 5-year follow-up period, 127 persons (73 men and 54 women) died. In Cox multivariate proportional hazards models adjusted for gender, age, number of years of education, living alone, presence of psychiatric problems, and presence of chronic diseases, conscientiousness (risk ratio [RR] = 0.50, 95% confidence interval [CI] = 0.33-0.76 and RR = 0.46, 95% CI = 0.29 to 0.71, for the middle and highest tertiles, respectively), extraversion (RR = 0.58, 95% CI = 0.37 to 0.92, for the highest tertile), and openness (RR = 0.62, 95% CI = 0.41 to 0.96, for the middle tertile) were independently and inversely associated with all-cause mortality. Neither neuroticism nor agreeableness was related to mortality. CONCLUSIONS: Our results suggest that out of the five domains of personality traits, conscientiousness, extraversion, and openness are reliable predictors of all-cause mortality among community-dwelling elderly. PMID- 18403572 TI - Facial emotion recognition deficit in amnestic mild cognitive impairment and Alzheimer disease. AB - OBJECTIVES: A deficit in facial emotion recognition was described in patients with Alzheimer disease (AD). However, this issue has been underexplored in subjects with amnestic mild cognitive impairment (a-MCI). Thus, the authors aimed to determine whether a deficit in facial emotion recognition is present in a-MCI phase and whether this is intensity dependent. A secondary aim was to investigate relationships between facial emotion recognition and cognitive performances. DESIGN: Case-control study. SETTING: Memory clinic. PARTICIPANTS: Fifty a-MCI patients, 50 mild AD patients, and 50 comparison subjects (COM) were enrolled. MEASUREMENTS: Information about facial emotion recognition was obtained from Penn Emotion Recognition Test. The Mental Deterioration Battery was used to measure cognitive impairment. RESULTS: Mild AD patients were more impaired in the recognition of almost all emotional stimuli of all intensities than a-MCI and COM subjects. However, there was an increased progression only in low-intensity facial emotion recognition deficit from COM to a-MCI to mild AD patients. In particular, a-MCI subjects differed significantly from COM in low-intensity fearful face recognition performance. This deficit in a-MCI patients was explained by the short-term verbal memory impairment, whereas the same deficit in mild AD patients was explained by the long-term verbal memory impairment. CONCLUSIONS: Emotion recognition progresses from a deficit in low-intensity fearful facial recognition in a-MCI phase to a deficit in all intensities and emotions in mild AD. This could be an effect of the progressive degeneration of brain structures modulating emotional processing. An early detection of emotional impairment in MCI phases of dementia may have clinical implications. PMID- 18403573 TI - Nurses' perceptions of hospice palliative care volunteers. AB - A total of 50 nurses (hospital and home care) responded to a survey designed to measure: (1) their attitudes toward, and knowledge of, hospice palliative care volunteers; (2) the types of tasks they felt it was appropriate for volunteers to perform; and (3) how valuable they felt different members of the hospice palliative care team are. In addition, they were asked to respond to some open ended questions (eg, "Do you feel that it is appropriate for hospice palliative care volunteers to know patient medical information?"). The nurses' responses to the "Attitude/Knowledge" part of the survey revealed that they generally held positive attitudes toward volunteers. The majority of the nurses felt that it was appropriate for volunteers to perform most of the tasks listed, except for hands on patient care. Nurses rated the value of nurses, family members, doctors, and pharmacists significantly higher than volunteers. Fifty-three percent of the nurses felt that volunteers should know patient medical information, and 77% thought that volunteers should have the opportunity to provide input regarding patient care. Also, 75% of the nurses felt that volunteers made their jobs easier, and 56% felt that volunteers should be included in team meetings. When asked to list the topics covered in a hospice palliative care volunteer training program, 73% of the nurses indicated that they were not sure or did not know what topics were covered, indicating a lack of knowledge regarding volunteer training. PMID- 18403574 TI - Destroying myths. AB - Palliative care emergencies such as spinal cord compression require prompt diagnosis and treatment to get the best results. Hospitals dealing with these emergencies need to develop pathways of care for these patients based on best evidence derived from hard data. The authors looked at all spinal cord compressions that presented to their hospital over a 10-year period. The authors found that the commonest day of presentation of this condition was on Mondays and not Fridays as had been widely believed prior to this study. Hospitals are encouraged to look at hard data and not anecdotes when developing pathways of care for their patients. PMID- 18403575 TI - No relief in sight: postgraduate training in pain management. AB - This study investigated training in pain management in postgraduate medical education programs. A mail survey of program directors was conducted, evaluating the format of training in pain management and the self-assessed adequacy of the training. The response rate was 70%, with 188 total respondents. It included all programs in a large Midwestern state, representing most specialties. Just over half of all programs offered any formal training in pain management, and even fewer required it. Less than one quarter required a clinical component to such training. Nonetheless, an overwhelming majority of program directors (85%) rated their training as adequate or excellent. Improvements are clearly needed in postgraduate training in pain management, and external incentives, such as requirements in the accreditation process, will be needed. PMID- 18403576 TI - Use of advance directives for nursing home residents in the emergency department. AB - Documented requests can ensure that patients' end-of-life care preferences are implemented, particularly in emergent circumstances. This study a) compared information on advance directives found in different sources of documentation in the hospital record of nursing home patients admitted through the emergency department and b) assessed emergency department clinicians' perceptions of how end-of-life care requests are communicated to them. Seven potential sources of documentation were reviewed in the medical records of 40 patients, and semistructured interviews were conducted with 10 emergency department clinicians. We found little concordance among sources of advance directive documentation. Our results suggest variability in documentation for nursing home patients on transfer to the emergency department, and that emergency department clinicians experience substantial difficulty in reliably obtaining information about advance directives. As treatment may vary based solely on available documentation, such information gaps may decrease the likelihood of adherence in the emergency department to patients' previously expressed care preferences. PMID- 18403577 TI - Cytokines and cancer anorexia cachexia syndrome. AB - Cancer anorexia cachexia syndrome is frequent yet still a not well understood cancer-related problem. The pathophysiology of cancer cachexia is multifactorial. It is suggested to be the result of tumor-host interactions and studies of the disturbances seen during cancer anorexia cachexia syndrome, such as anorexia, hyper-metabolism, tissue wasting, metabolic abnormalities, and hormonal changes, all point to the involvement in one way or another of one key factor: cytokines. The purpose of this review is to summarize the latest developments in the field of cytokines and their role in cancer anorexia cachexia syndrome. The emphasis is on the role of cytokines in anorexia and tissue wasting. PMID- 18403578 TI - Using the LCP: bereaved relatives' assessments of communication and bereavement. AB - The Liverpool Care Pathway (LCP) is aimed at improving care and communication in the dying phase. The authors studied whether use of the LCP affects relatives' retrospective evaluation of communication and their level of bereavement. An intervention study was conducted. During the baseline period, usual care was provided to dying patients. During the intervention period, the LCP was used for 79% of the patients. In total, bereaved relatives filled in a questionnaire for 57% of the patients, on average 4 months after death. In the intervention period, relatives had lower bereavement levels when compared with relatives in the baseline period (P = .01). Communication was evaluated similarly for both periods. We conclude that LCP use during the dying phase seems to moderately contribute to lower levels of bereavement in relatives. PMID- 18403579 TI - Is head balance a major determinant for swallowing problems in patients with spinal muscular atrophy type 2? AB - A child with spinal muscular atrophy type 2 was referred for evaluation of eating and swallowing problems. The dysphagia evaluation demonstrated coughing during eating and drinking and occasionally stertorous when eating solid food. The videofluoroscopic swallow study showed a late upper esophageal sphincter opening with hypopharyngeal residue, more with solid food than with thin liquid. His lumbar lordosis associated with anterior tilted pelvis and his problems with head balance due to weak neck musculature caused compensatory behavior like a retracted neck and mandible. This position negatively influences the opening of the upper esophageal sphincter. This case supported the idea that dysphagia in spinal muscular atrophy type 2 is caused by both a bulbar component as well as a treatable posture component. PMID- 18403580 TI - Ataxia with oculomotor apraxia type 1 (AOA1): clinical and neuropsychological features in 2 new patients and differential diagnosis. AB - Ataxia with oculomotor apraxia type 1 (AOA1) is an autosomal recessive disease characterized by early-onset and slowly progressive cerebellar ataxia, areflexia, and peripheral neuropathy. Ocular apraxia is most prominent in the early stage of the disease, by contrast, hypoalbuminemia, hypercholesterolemia, and cognitive impairment are present in the adult stage. AOA1 is caused by a mutation in the APTX gene (9p13.3) encoding a nuclear protein named aprataxin, which is involved in the mechanism of DNA repair. We report here the clinical features of 2 patients with mutations in the APTX gene, and we discuss the differential diagnosis with other forms of hereditary ataxia. PMID- 18403581 TI - An unusual manifestation of a neonatal Chlamydia infection. AB - A 25-day-old boy was admitted to hospital because of pneumonia and additionally developed symptoms of encephalitis. The immune fluorescence test for Chlamydia trachomatis in tracheal fluids was positive. Furthermore, ligase chain reaction for C trachomatis was positive in the cerebrospinal fluid. The antibiotic regimen was changed to erythromycin intravenously. C trachomatis encephalitis is rare in neonates and may result from a defect in the alternative pathway of complement activation which was the case in this patient. PMID- 18403582 TI - Health care utilization and expenditures for children and young adults with muscular dystrophy in a privately insured population. AB - We provide estimates of medical care utilization and expenditures for children and young adults younger than age 30 with muscular dystrophies in the United States. Accurate estimates are essential for calculations of lifetime costs and for economic evaluations of screening and management strategies for muscular dystrophy. We compare the medical expenditures for persons with muscular dystrophy with others by age groups. The incremental annual expenditures of medical care for privately insured individuals with muscular dystrophy relative to others in 2004 averaged $18 930 and ranged from $13 464 at ages 5 to 9 to $32 541 at ages 15 to 19. Individuals with muscular dystrophy had average medical expenditures 10 to 20 times greater than individuals without muscular dystrophy. Individuals aged 15 to 19 years had the highest number of inpatient admissions related to respiratory infections and cardiac complications. The findings underscore the need for appropriate treatment options for individuals with muscular dystrophy as they age. PMID- 18403583 TI - GLUT1 deficiency without epilepsy: yet another case. AB - Glucose transporter type 1 (GLUT1) deficiency syndrome is a metabolic disorder characterized by a low cerebrospinal fluid glucose level caused by decreased activity of the glucose transporter protein. Of approximately 100 patients described with this syndrome in the published literature to date, only 3 patients have had intermittent ataxia as the initial manifestation. This case report describes a 13-year-old boy with a longstanding history of intermittent ataxia who was diagnosed as having GLUT1 deficiency syndrome after the onset of seizures at age 11 years. This case highlights the importance of a carefully organized lumbar puncture in the investigation and management of any child with neurodevelopmental delay and intermittent ataxia with or without seizures. PMID- 18403584 TI - Griscelli syndrome type 2: a rare and lethal disorder. AB - Griscelli syndrome is a rare autosomal recessive disorder. It is characterized by pigment dilution and variable immune deficiency leading to increased susceptibility to certain infections and a tendency to develop a life-threatening hemophagocytic syndrome known as the accelerated phase. Griscelli syndrome is now classified into 3 types based on the genetic and molecular features. Primary neurological presentation without the accelerated phase is rare in type 2. In this article, the authors report a boy who was presented with seizures and diffuse white matter involvement unaccompanied by the other features of the accelerated phase. Mutation analysis in family members revealed the presence of a missense mutation in Rab27a gene. In addition to the rare presentation, this is the first case of Griscelli syndrome to be reported from Jordan. PMID- 18403585 TI - Giant infantile gliosarcoma: magnetic resonance imaging findings. AB - Gliosarcoma is an uncommon variant of glioblastoma multiforme, which is composed of gliomatous and sarcomatous elements. The tumor is rarely encountered in childhood. This case report presents the magnetic resonance imaging characteristics of a giant gliosarcoma in a 3-year-old girl. Size and location of the tumor are described. PMID- 18403586 TI - Decompressive hemicraniectomy in children with severe ischemic stroke and life threatening cerebral edema. AB - Decompressive hemicraniectomy has been discussed as a treatment option that increases survival in adults with malignant stroke. This approach has not been studied extensively in children. From a prospective cohort, we identified 4 children who underwent decompressive hemicraniectomy for malignant infarctions with life-threatening cerebral edema within 72 hours of their stroke. All 4 children had different causes for their stroke and experienced severe cerebral edema with increasing intracranial pressure and an impending fatal outcome. Despite massive cerebral infarction, all patients were ambulant and able to speak at the time of follow-up. Although a limited experience, decompressive hemicraniectomy is a life-saving approach for malignant stroke in children. PMID- 18403587 TI - Development of diabesity in mice with neuronal deletion of Shp2 tyrosine phosphatase. AB - Obesity and diabetes, termed "diabesity," are serious health problems that are increasing in frequency. However, the molecular mechanisms and neuronal regulation of these metabolic disorders are not fully understood. We show here that Shp2, a widely expressed Src homology 2-containing Tyr phosphatase, plays a critical role in the adult brain to control food intake, energy balance, and metabolism. Mice with a neuron-specific, conditional Shp2 deletion were generated by crossing a pan-neuronal Cre-line (CRE3) with Shp2(flox/flox) mice. These congenic mice, CRE3/Shp2-KO, developed obesity and diabetes and the associated pathophysiological complications that resemble those encountered in humans, including hyperglycemia, hyperinsulinemia, hyperleptinemia, insulin and leptin resistance, vasculitis, diabetic nephropathy, urinary bladder infections, prostatitis, gastric paresis, and impaired spermatogenesis. This mouse model may help to elucidate the molecular mechanisms that lead to the development of diabesity in humans and provide a tool to study the in vivo complications of uncontrolled diabetes. PMID- 18403588 TI - Neuropeptide modulators of high mobility group box 1 secretion as potential therapeutic agents for severe sepsis. PMID- 18403589 TI - Topical thymidine dinucleotide treatment reduces development of ultraviolet induced basal cell carcinoma in Ptch-1+/- mice. AB - Treatment with thymidine dinucleotide (pTT) has well documented DNA-protective effects and reduces development of squamous cell carcinoma in UV-irradiated mice. The preventive effect of pTT on basal cell carcinoma (BCC) was evaluated in UV irradiated Ptch-1(+/-) mice, a model of the human disease Gorlin syndrome. Topical pTT treatment significantly reduced the number and size (P < 0.001) of BCCs in murine skin after 7 months of chronic irradiation. Skin biopsies collected 24 hours after the final UV exposure showed that pTT reduced the number of nuclei positive for cyclobutane pyrimidine dimers by 40% (P < 0.0002) and for 8-hydroxy-2'-deoxyguanosine by 61% (P < 0.01 compared with vehicle control). Immunostaining with an antibody specific for mutated p53 revealed 63% fewer positive patches in BCCs of pTT-treated mice compared with controls (P < 0.01), and the number of Ki-67-positive cells was decreased by 56% (P < 0.01) in pTT treated tumor-free epidermis and by 76% (P < 0.001) in BCC tumor nests (P < 0.001). Terminal dUTP nick-end labeling staining revealed a 213% increase (P < 0.04) in the number of apoptotic cells in BCCs of pTT-treated mice. Cox-2 immunostaining was decreased by 80% in tumor-free epidermis of pTT-treated mice compared with controls (P < 0.01). We conclude that topical pTT treatment during a prolonged period of intermittent UV exposure decreases the number and size of UV-induced BCCs through several anti-cancer mechanisms. PMID- 18403590 TI - Neprilysin: an enzyme candidate to slow the progression of Alzheimer's disease. AB - It is well established that the extracellular deposition of amyloid beta (Abeta) peptide plays a central role in the development of Alzheimer's disease (AD). Therefore, either preventing the accumulation of Abeta peptide in the brain or accelerating its clearance may slow the rate of AD onset. Neprilysin (NEP) is the dominant Abeta peptide-degrading enzyme in the brain; NEP becomes inactivated and down-regulated during both the early stages of AD and aging. In this study, we investigated the effect of human (h)NEP gene transfer to the brain in a mouse model of AD before the development of amyloid plaques, and assessed how this treatment modality affected the accumulation of Abeta peptide and associated pathogenetic changes (eg, inflammation, oxidative stress, and memory impairment). Overexpression of hNEP for 4 months in young APP/DeltaPS1 double-transgenic mice resulted in reduction in Abeta peptide levels, attenuation of amyloid load, oxidative stress, and inflammation, and improved spatial orientation. Moreover, the overall reduction in amyloidosis and associated pathogenetic changes in the brain resulted in decreased memory impairment by approximately 50%. These data suggest that restoring NEP levels in the brain at the early stages of AD is an effective strategy to prevent or attenuate disease progression. PMID- 18403591 TI - Diabetes-enhanced tumor necrosis factor-alpha production promotes apoptosis and the loss of retinal microvascular cells in type 1 and type 2 models of diabetic retinopathy. AB - Retinal microvascular cell loss plays a critical role in the pathogenesis of diabetic retinopathy. To examine this further, type 1 streptozotocin-induced diabetic rats and type 2 Zucker diabetic fatty rats were treated by intravitreal injection of the tumor necrosis factor-specific inhibitor pegsunercept, and the impact was measured by analysis of retinal trypsin digests. For type 2 diabetic rats, the number of endothelial cells and pericytes positive for diabetes enhanced activated caspase-3 decreased by 81% and 86%, respectively, when treated with pegsunercept (P < 0.05). Similarly, the number of diabetes-enhanced terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive endothelial cells and pericytes decreased by 81% and 67% respectively when treated with pegsunercept (P < 0.05). Diabetes-increased activated caspase-3- and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive microvascular cell numbers were both reduced by 81% and 80%, respectively, in pegsunercept-treated type 1 diabetic rats (P < 0.05). Inhibition of tumor necrosis factor reduced type 1 diabetes-enhanced pericyte ghost formation by 87% and the number of type 2 diabetes-enhanced pericyte ghosts by 62% (P < 0.05). Similarly, increased acellular capillary formation caused by type 1 and type 2 diabetes was reduced by 68% and 67%, respectively, when treated with pegsunercept (P < 0.05). These results demonstrate a previously unrecognized role of tumor necrosis factor-alpha in promoting the early pathogenesis of diabetic retinopathy leading to loss of retinal microvascular cells and demonstrate the potential therapeutic benefit of modulating its activity. PMID- 18403592 TI - Molecular and cytogenetic characterization of plexiform leiomyomata provide further evidence for genetic heterogeneity underlying uterine fibroids. AB - The plexiform variant of uterine leiomyomata (UL) is named for its ribbons or nests of smooth muscle cells that have a rounded, epithelioid shape caused by their entrapment in abundant extracellular matrix. Plexiform UL are currently classified as epithelioid smooth muscle tumors alongside the less predictable, "true" epithelioid tumors (ie, leiomyoblastomas). Karyotypes of six plexiform UL cases were studied, and their abnormalities were found to differ from those of leiomyoblastomas. Analyses using real-time polymerase chain reaction, immunohistochemistry, and fluorescence in situ hybridization demonstrated elevated mRNA and protein levels of the architectural factor HMGA2 and, in some cases, increased DNA copy number. Four of these plexiform UL were profiled with Affymetrix human U133 plus 2.0 expression arrays. Cluster analysis using genes previously shown to discriminate benign and malignant uterine smooth muscle tissues revealed that the plexiform tumors form an isolated group in the benign branch. This is in contrast to an earlier finding in which another variant, cellular UL characterized by loss of a portion of the short arm of chromosome 1, clustered with malignant leiomyosarcomas. These results provide additional evidence of genetic heterogeneity underlying UL of various histological types. We further suggest that plexiform UL should be classified among tumors with extensive hyalinization rather than with "true" epithelioid smooth muscle neoplasms. PMID- 18403593 TI - The benefit of docosahexanoic acid on the migration of vascular smooth muscle cells is partially dependent on Notch regulation of MMP-2/-9. AB - The Notch pathway is involved in the regulation of the migratory/proliferative phenotype acquired by vascular smooth muscle cells (VSMCs) in the pro inflammatory context of vascular diseases. Here, we investigated whether docosahexaenoic acid (DHA), a polyunsaturated, omega-3 fatty acid, could reduce fibrinolytic/matrix-metalloproteinase (MMP) activity and whether this reduction occurs through the modulation of Notch signaling. Rat VSMCs were transdifferentiated with interleukin-1beta and then treated with DHA. Migration/proliferation was determined by performing a wound healing assay and measuring MMP-2/-9 activity, type 1 plasminogen activator inhibitor levels, and the expression of these proteins. The involvement of Notch in regulating the fibrinolytic/MMP system was evidenced using Notch pathway inhibitors and the forced expression of Notch1 and Notch3 intracellular domains. DHA significantly decreased VSMC migration/proliferation induced by interleukin-1beta as well as fibrinolytic/MMP activity. Prevention of Notch1 target gene transcription enhanced the interleukin-1beta effects on MMPs and on migration, whereas Notch3 intracellular domain overexpression reduced these effects. Finally, DHA increased Notch3 expression, Hes-1 transcription (a Notch target gene), and enhanced gamma secretase complex activity. These results suggest that inhibition of the Notch pathway participates in the transition of VSMCs toward a migratory phenotype. These results also suggest that the beneficial inhibitory effects of DHA on fibrinolytic/MMP activity are related in part to the effects of DHA on the expression of Notch pathway components, providing new insight into the mechanisms by which omega-3 fatty acids prevent cardiovascular diseases. PMID- 18403594 TI - Molecular characterization of human breast tumor vascular cells. AB - A detailed understanding of the assortment of genes that are expressed in breast tumor vessels is needed to facilitate the development of novel, molecularly targeted anti-angiogenic agents for breast cancer therapies. Rapid immunohistochemistry using factor VIII-related antibodies was performed on sections of frozen human luminal-A breast tumors (n = 5) and normal breast (n = 5), followed by laser capture microdissection of vascular cells. RNA was extracted and amplified, and fluorescently labeled cDNA was synthesized and hybridized to 44,000-element long-oligonucleotide DNA microarrays. Statistical analysis of microarray was used to compare differences in gene expression between tumor and normal vascular cells, and Expression Analysis Systematic Explorer was used to determine enrichment of gene ontology categories. Protein expression of select genes was confirmed using immunohistochemistry. Of the 1176 genes that were differentially expressed between tumor and normal vascular cells, 55 had a greater than fourfold increase in expression level. The extracellular matrix gene ontology category was increased while the ribosome gene ontology category was decreased. Fibroblast activation protein, secreted frizzled-related protein 2, Janus kinase 3, and neutral sphingomyelinase 2 proteins localized to breast tumor endothelium as assessed by immunohistochemistry, showing significantly greater staining compared with normal tissue. These tumor endothelial marker proteins also exhibited increased expression in breast tumor vessels compared with that in normal tissues. Therefore, these genetic markers may serve as potential targets for the development of angiogenesis inhibitors. PMID- 18403595 TI - Angiotensin II up-regulates angiotensin I-converting enzyme (ACE), but down regulates ACE2 via the AT1-ERK/p38 MAP kinase pathway. AB - The recent discovery of the angiotensin II (Ang II)-breakdown enzyme, angiotensin I converting enzyme (ACE) 2, suggests the importance of Ang II degradation in hypertension. The present study explored the signaling mechanism by which ACE2 is regulated under hypertensive conditions. Real-time PCR and immunohistochemistry showed that ACE2 mRNA and protein expression levels were high, whereas ACE expression levels were moderate in both normal kidney and heart. In contrast, patients with hypertension showed marked ACE up-regulation and ACE2 down regulation in both hypertensive cardiopathy and, particularly, hypertensive nephropathy. The inhibition of ACE2 expression was shown to be associated with ACE up-regulation and activation of extracellular regulated (ERK)1/2 and p38 mitogen-activated protein (MAP) kinases. In vitro, Ang II was able to up-regulate ACE and down-regulate ACE2 in human kidney tubular cells, which were blocked by an angiotensin II (AT)1 receptor antagonist (losartan), but not by an AT2 receptor blocker (PD123319). Furthermore, blockade of ERK1/2 or p38 MAP kinases by either specific inhibitors or a dominant-negative adenovirus was able to abolish Ang II-induced ACE2 down-regulation in human kidney tubular cells. In conclusion, Ang II is able to up-regulate ACE and down-regulate ACE2 expression levels under hypertensive conditions both in vivo and in vitro. The AT1 receptor mediated ERK/p38 MAP kinase signaling pathway may be a key mechanism by which Ang II down-regulates ACE2 expression, implicating an ACE/ACE2 imbalance in hypertensive cardiovascular and renal damage. PMID- 18403596 TI - Cross-species comparison of human and mouse intestinal polyps reveals conserved mechanisms in adenomatous polyposis coli (APC)-driven tumorigenesis. AB - Expression profiling is a well established tool for the genome-wide analysis of human cancers. However, the high sensitivity of this approach combined with the well known cellular and molecular heterogeneity of cancer often result in extremely complex expression signatures that are difficult to interpret functionally. The majority of sporadic colorectal cancers are triggered by mutations in the adenomatous polyposis coli (APC) tumor suppressor gene, leading to the constitutive activation of the Wnt/beta-catenin signaling pathway and formation of adenomas. Despite this common genetic basis, colorectal cancers are very heterogeneous in their degree of differentiation, growth rate, and malignancy potential. Here, we applied a cross-species comparison of expression profiles of intestinal polyps derived from hereditary colorectal cancer patients carrying APC germline mutations and from mice carrying a targeted inactivating mutation in the mouse homologue Apc. This comparative approach resulted in the establishment of a conserved signature of 166 genes that were differentially expressed between adenomas and normal intestinal mucosa in both species. Functional analyses of the conserved genes revealed a general increase in cell proliferation and the activation of the Wnt/beta-catenin signaling pathway. Moreover, the conserved signature was able to resolve expression profiles from hereditary polyposis patients carrying APC germline mutations from those with bi allelic inactivation of the MYH gene, supporting the usefulness of such comparisons to discriminate among patients with distinct genetic defects. PMID- 18403597 TI - Genetic ablation of NADPH oxidase enhances susceptibility to cigarette smoke induced lung inflammation and emphysema in mice. AB - Cigarette smoke (CS) induces recruitment of inflammatory cells in the lungs leading to the generation of reactive oxygen species (ROS), which are involved in lung inflammation and injury. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is a multimeric system that is responsible for ROS production in mammalian cells. We hypothesized that NADPH oxidase-derived ROS play an important role in lung inflammation and injury and that targeted ablation of components of NADPH oxidase (p47(phox) and gp91(phox)) would protect lungs against the detrimental effects of CS. To test this hypothesis, we exposed p47(phox-/-) and gp91(phox-/-) mice to CS and examined inflammatory response and injury in the lung. Surprisingly, although CS-induced ROS production was decreased in the lungs of p47(phox-/-) and gp91(phox-/-) mice compared with wild-type mice, the inflammatory response was significantly increased and was accompanied by development of distal airspace enlargement and alveolar destruction. This pathological abnormality was associated with enhanced activation of the TLR4 nuclear factor-kappaB pathway in response to CS exposure in p47(phox-/-) and gp91(phox-/-) mice. This phenomenon was confirmed by in vitro studies in which treatment of peritoneal macrophages with a nuclear factor-kappaB inhibitor reversed the CS-induced release of proinflammatory mediators. Thus, these data suggest that genetic ablation of components of NADPH oxidase enhances susceptibility to the proinflammatory effects of CS leading to airspace enlargement and alveolar damage. PMID- 18403599 TI - To matrigel or not to matrigel. PMID- 18403598 TI - Interleukin-13 protects against experimental autoimmune myocarditis by regulating macrophage differentiation. AB - We report here that interleukin (IL)-13 protects BALB/c mice from myocarditis, whether induced by peptide immunization or by viral infection. In contrast to mild disease in IL-4 knockout (KO) BALB/c mice, IL-13 KO BALB/c mice developed severe coxsackievirus B3 (CVB3)-induced autoimmune myocarditis and myocarditogenic peptide-induced experimental autoimmune myocarditis. Such severe disease was characterized by increased cardiac inflammation, increased total intracardiac CD45(+) leukocytes, elevated anti-cardiac myosin autoantibodies, and increased cardiac fibrosis. Echocardiography revealed that IL-13 KO mice developed severe dilated cardiomyopathy with impaired cardiac function and heart failure. Hearts of IL-13 KO mice had increased levels of the proinflammatory and profibrotic cytokines IL-1beta, IL-18, interferon-gamma, transforming growth factor-beta1, and IL-4 as well as histamine. The hallmark of the disease in IL-13 KO mice was the up-regulation of T-cell responses. CD4(+) T cells were increased in IL-13 KO hearts both proportionally and in absolute number. Splenic T cells from IL-13 KO mice were highly activated, and myosin stimulation additionally increased T-cell proliferation. CD4(+)CD25(+)Foxp3(+) regulatory T-cell numbers were decreased in the spleens of IL-13 KO mice. IL-13 deficiency led to decreased levels of alternatively activated CD206(+) and CD204(+) macrophages and increased levels of classically activated macrophages. IL-13 KO mice had increased caspase 1 activation, leading to increased production of both IL-1beta and IL-18. Therefore, IL-13 protects against myocarditis by modulating monocyte/macrophage populations and by regulating their function. PMID- 18403600 TI - A novel mechanism for CCR4 in the regulation of macrophage activation in bleomycin-induced pulmonary fibrosis. AB - Macrophage polarization into M1 or M2 phenotypes dictates the nature, duration, and severity of an inflammatory response. The objective of this study was to examine the role of CC chemokine receptor 4 (CCR4) in macrophage polarization during pulmonary oxidative injury in wild-type [WT (CCR4(+/+))] and CCR4 deficient (CCR4(-/-)) mice. Intrapulmonary administration of bleomycin sulfate provoked lethal inflammatory and fibrotic responses in WT (CCR4(+/+)) mice, but such responses were absent in CCR4(-/-) mice. Transcript and protein analyses of alveolar and bone marrow-derived macrophages showed that cells isolated from CCR4(-/-) mice did not exhibit CCL17-dependent M1 activation in response to bleomycin. Instead, CCR4(-/-) macrophages showed an M2 phenotype characterized by significantly elevated expression of arginase 1 and FIZZ1 (found in inflammatory zone 1), particularly during the peak of pulmonary inflammation. Compared with WT (CCR4(+/+)) mice, CCR4(-/-) mice exhibited a significant increase in the expression of the nonsignaling CC chemokine scavenging receptor D6 in whole lung samples and isolated macrophages. Thus, these results demonstrate that CCL17 dependent activation of CCR4 in macrophages plays a central role in free radical induced pulmonary injury and repair. PMID- 18403602 TI - Matrix metalloproteinase 12 accelerates the initiation of atherosclerosis and stimulates the progression of fatty streaks to fibrous plaques in transgenic rabbits. AB - Whether fatty streaks are directly followed by fibrous plaque formation in atherosclerosis remains controversial. Disruption of the basement membrane and elastic layers is thought to be essential for this process. Matrix metalloproteinase 12 (MMP-12) can degrade a broad spectrum of substrates, but the role of MMP-12 in the early stage of atherosclerosis is unclear. To investigate MMP-12 function in the initiation and progression of atherosclerosis, we investigated macrophage migration and elastolysis in relation to fatty streaks in human MMP-12 transgenic (hMMP-12 Tg) rabbits. Fatty streaks in hMMP-12 Tg rabbits fed a 1% cholesterol diet for 6 weeks (cholesterol-induced model of atherosclerosis) were more pronounced and were associated with more significant degradation of the internal elastic layer compared with wild-type (WT) animals. Numbers of infiltrating macrophages and smooth muscle cells in the lesions were increased in hMMP-12 Tg compared with WT animals. In both cuff- and ligation induced models of atherosclerosis, smooth muscle cell-predominant atherosclerotic lesions were elevated with significant elastolysis of the internal elastic lamina in Tg compared with WT animals; "microelastolytic sites" were recognized before formation of the neointima in the cuff model only. These results indicate that MMP-12 may be critical to the initiation and progression of atherosclerosis via degradation of the elastic layers and/or basement membrane. Therefore, a specific MMP-12 inhibitor might prove useful for the treatment of progressive atherosclerosis. PMID- 18403601 TI - Regulated intramembrane proteolysis of the low-density lipoprotein receptor related protein mediates ischemic cell death. AB - The low-density lipoprotein receptor-related protein (LRP), a member of the low density lipoprotein receptor gene family, mediates cellular signal transduction pathways. In this study we investigated the role of LRP in cell death. We found that incubation of mouse embryonic fibroblasts in serum-free media induces caspase-3 activation, an effect that is attenuated in LRP-deficient (LRP(-/-)) mouse embryonic fibroblasts. Since we previously demonstrated that middle cerebral artery occlusion (MCAO) in mice induces shedding of the LRP ectodomain, we investigated here whether cerebral ischemia induces regulated intramembrane proteolysis of LRP and whether this process is related to cell death. We found that MCAO induces an increase in gamma-secretase activity in the ischemic hemisphere and that treatment with the gamma-secretase inhibitor L-685,458 improves the neurological outcome and results in a 50% decrease in the volume of the ischemic lesion. Furthermore, MCAO caused nuclear translocation of the intracellular domain of LRP in neurons within the area of ischemic penumbra, and this effect was attenuated in mice treated with L-685,458. Finally, inhibition of either LRP or gamma-secretase attenuated cerebral ischemia-induced caspase-3 cleavage and apoptotic cell death. In summary, our results indicate that gamma secretase-mediated regulated intramembrane proteolysis of LRP results in cell death under ischemic conditions. PMID- 18403603 TI - Signaling pathways for B cell-activating factor (BAFF) and a proliferation inducing ligand (APRIL) in human placenta. AB - The tumor necrosis superfamily (TNFSF) contains two soluble ligands that are involved in B lymphocyte development, BAFF (B cell activating factor, BlyS, TALL 1, CD257, TNFSF13B) and APRIL (a proliferation inducing ligand, CD256, TNFSF13). These two ligands signal through three receptors: the exclusive BAFF receptor (BAFF-R, CD268, TNFRSF17) and two receptors that recognize both BAFF and APRIL, TACI (transmembrane-activator-1 and calcium-modulator- and cyclophilin ligand interactor CD267, TNFRSF13B) and BCMA (B cell maturation antigen, CD269, TNFRSF13C). All but BAFF-R are known to be synthesized in term placentas. In this study, expression of the ligands and receptors were distinguished in two embryologically discrete subpopulations of placental cells, villous cytotrophoblast (vCTB) cells and mesenchymal cells (MCs). Real-Time PCR showed that vCTB cells contain low levels of BAFF and APRIL transcripts whereas MCs contain high levels. Both Real-Time PCR and immunohistochemistry identified BAFF R and BCMA mRNA and proteins in vCTB cells but essentially no TACI. By contrast, MCs contained readily detectable levels of all three receptors. These results illustrating potential autocrine and paracrine pathways for BAFF and APRIL signaling in human placentas suggest that lineage-specific regulation of placental cell viability, differentiation and/or other activities may be novel functions of these proteins. PMID- 18403605 TI - A new DNA-based test for detection of nucleophosmin exon 12 mutations by capillary electrophoresis. AB - Mutations in nucleophosmin (NPM1) exon 12 are thought to be the most common genetic event in acute myelogenous leukemia (AML) and to confer favorable clinical prognoses. In this report, we describe a simple molecular test for the detection of NPM1 exon 12 mutations in patients with AML using polymerase chain reaction amplification of genomic DNA followed by the analysis of amplification products by capillary electrophoresis. Mutations were reproducibly detected when present in at least 5% of cells, and all NPM1 exon 12 mutations reported to date in AML could be identified using this method. This method was successfully employed using paraffin-extracted DNA, allowing for the examination of archived clinical specimens, and the assay was validated by the direct sequencing of 33 patient samples. This sensitive test is straightforward to perform and provides important information that can influence both the clinical management and treatment options for many patients with AML. PMID- 18403604 TI - Pathology, molecular biology, and pathogenesis of avian influenza A (H5N1) infection in humans. AB - H5N1 avian influenza is a highly fatal infectious disease that could cause a potentially devastating pandemic if the H5N1 virus mutates into a form that spreads efficiently among humans. Recent findings have led to a basic understanding of cell and organ histopathology caused by the H5N1 virus. Here we review the pathology of H5N1 avian influenza reported in postmortem and clinical studies and discuss the key pathogenetic mechanisms. Specifically, the virus infects isolated pulmonary epithelial cells and causes diffuse alveolar damage and hemorrhage in the lungs of infected patients. In addition, the virus may infect other organs, including the trachea, the intestines, and the brain, and it may penetrate the placental barrier and infect the fetus. Dysregulation of cytokines and chemokines is likely to be one of the key mechanisms in the pathogenesis of H5N1 influenza. We also review the various molecular determinants of increased pathogenicity that have been identified in recent years and the role of avian and human influenza virus receptors in relation to the transmissibility of the H5N1 virus. A comprehensive appreciation of H5N1 influenza pathogenetic mechanisms should aid in the design of effective strategies for prevention, diagnosis, and treatment of this emerging disease. PMID- 18403606 TI - Performance of whole-genome amplified DNA isolated from serum and plasma on high density single nucleotide polymorphism arrays. AB - Defining genetic variation associated with complex human diseases requires standards based on high-quality DNA from well-characterized patients. With the development of robust technologies for whole-genome amplification, sample repositories such as serum banks now represent a potentially valuable source of DNA for both genomic studies and clinical diagnostics. We assessed the performance of whole-genome amplified DNA (wgaDNA) derived from stored serum/plasma on high-density single nucleotide polymorphism arrays. Neither storage time nor usage history affected either DNA extraction or whole-genome amplification yields; however, samples that were thawed and refrozen showed significantly lower call rates (73.9 +/- 7.8%) than samples that were never thawed (92.0 +/- 3.3%) (P < 0.001). Genotype call rates did not differ significantly (P = 0.13) between wgaDNA from never-thawed serum/plasma (92.9 +/- 2.6%) and genomic DNA (97.5 +/- 0.3%) isolated from whole blood. Approximately 400,000 genotypes were consistent between wgaDNA and genomic DNA, but the overall discordance rate of 4.4 +/- 3.8% reflected an average of 11,110 +/- 9502 genotyping errors per sample. No distinct patterns of chromosomal clustering were observed for single nucleotide polymorphisms showing discordant genotypes or homozygote conversion. Because the effects of genotyping errors on whole-genome studies are not well defined, we recommend caution when applying wgaDNA from serum/plasma to high-density single nucleotide polymorphism arrays in addition to the use of stringent quality control requirements for the resulting genotype data. PMID- 18403607 TI - Direct sequence detection of structured h5 influenza viral RNA. AB - We describe the development of sequence-specific molecular beacons (dual-labeled DNA probes) for identification of the H5 influenza subtype, cleavage motif, and receptor specificity when hybridized directly with in vitro transcribed viral RNA (vRNA). The cloned hemagglutinin segment from a highly pathogenic H5N1 strain, A/Hanoi/30408/2005(H5N1), isolated from humans was used as template for in vitro transcription of sense-strand vRNA. The hybridization behavior of vRNA and a conserved subtype probe was characterized experimentally by varying conditions of time, temperature, and Mg2+ to optimize detection. Comparison of the hybridization rates of probe to DNA and RNA targets indicates that conformational switching of influenza RNA structure is a rate-limiting step and that the secondary structure of vRNA dominates the binding kinetics. The sensitivity and specificity of probe recognition of other H5 strains was calculated from sequence matches to the National Center for Biotechnology Information influenza database. The hybridization specificity of the subtype probes was experimentally verified with point mutations within the probe loop at five locations corresponding to the other human H5 strains. The abundance frequencies of the hemagglutinin cleavage motif and sialic acid recognition sequences were experimentally tested for H5 in all host viral species. Although the detection assay must be coupled with isothermal amplification on the chip, the new probes form the basis of a portable point-of-care diagnostic device for influenza subtyping. PMID- 18403608 TI - Emerging pathogens: challenges and successes of molecular diagnostics. AB - More than 50 emerging and reemerging pathogens have been identified during the last 40 years. Until 1992 when the Institute of Medicine issued a report that defined emerging infectious diseases, medicine had been complacent about such infectious diseases despite the alarm bells of infections with human immunodeficiency virus. Molecular tools have proven useful in discovering and characterizing emerging viruses and bacteria such as Sin Nombre virus (hantaviral pulmonary syndrome), hepatitis C virus, Bartonella henselae (cat scratch disease, bacillary angiomatosis), and Anaplasma phagocytophilum (human granulocytotropic anaplasmosis). The feasibility of applying molecular diagnostics to dangerous, fastidious, and uncultivated agents for which conventional tests do not yield timely diagnoses has achieved proof of concept for many agents, but widespread use of cost-effective, validated commercial assays has yet to occur. This review presents representative emerging viral respiratory infections, hemorrhagic fevers, and hepatitides, as well as bacterial and parasitic zoonotic, gastrointestinal, and pulmonary infections. Agent characteristics, epidemiology, clinical manifestations, and diagnostic methods are tabulated for another 22 emerging viruses and five emerging bacteria. The ongoing challenge to the field of molecular diagnostics is to apply contemporary knowledge to facilitate agent diagnosis as well as to further discoveries of novel pathogens. PMID- 18403609 TI - EGFR mutations in lung adenocarcinomas: clinical testing experience and relationship to EGFR gene copy number and immunohistochemical expression. AB - Lung adenocarcinomas responsive to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors possess EGFR mutations and often increased EGFR copy number. We prospectively studied 334 clinical cases using polymerase chain reaction-based assays to detect deletions within exon 19 and the L858R mutation in exon 21, which together account for approximately 90% of EGFR mutations. Seventy-eight (23%) of these tumors had an EGFR mutation, with 55 (71%) exon 19 deletions and 23 (29%) exon 21 L858R mutations. We were able to compare mutant and normal EGFR alleles and found a preferential amplification of the mutant allele. The association of mutations with EGFR amplification (determined by chromogenic in situ hybridization) and EGFR expression (determined by immunohistochemistry) was further examined in a subset of 60 tumors. EGFR amplification (> or =5 EGFR signals per nucleus) was seen in 15 of 29 (52%) EGFR mutated tumors but in only five of 31 (6%) non-mutated tumors (P = 0.006). EGFR overexpression was strongly associated with amplification but was statistically independent of EGFR mutation. Most patients with EGFR mutations (17 of 29, 59%) never smoked compared with 13% (four of 31) of patients lacking such mutations (P = 0.0003). The association of amplification with smoking status was marginal and was nonexistent with EGFR expression. Thus, these results indicate that EGFR amplification, preferentially of the mutant allele, often accompanies EGFR mutation, whereas EGFR immunohistochemical staining associates with amplification but cannot predict EGFR mutation status. PMID- 18403610 TI - Evaluation and validation of total RNA extraction methods for microRNA expression analyses in formalin-fixed, paraffin-embedded tissues. AB - Histopathology archives of well-annotated formalin-fixed, paraffin-embedded (FFPE) tissue specimens are valuable resources for retrospective studies of human diseases. Since recovery of quality intact mRNA compatible with molecular techniques is often difficult due to degradation, we evaluated microRNA (miRNA), a novel class of small RNA molecules with growing therapeutic and diagnostic potential, as an alternative analyte for gene expression studies of FFPE samples. Analyzing total RNA yield, miRNA recovery, and robustness of real-time polymerase chain reaction for miRNA, mRNA, and rRNA species, we compared the performance of commercially available RNA isolation kits and identified a preferred methodology. We further implemented modifications to increase tissue throughput and incorporate a quantitative Armored RNA process control to monitor RNA recovery efficiency. The optimized process was tested for reproducibility as well as interoperator and interday variability, and was validated with a set of 30 clinical samples. In addition, we demonstrated that, independent of FFPE block age and RNA quality, miRNAs generate quantitative reverse transcription polymerase chain reaction signals that are more robust and better correlate with expression levels from frozen reference samples compared with longer mRNAs. Our broad study, including a total of 272 independent RNA isolations from 17 tissue types and 65 FFPE blocks up to 12 years old, indicates that miRNAs are not only suitable but are also likely superior analytes for the molecular characterization of compromised archived clinical specimens. PMID- 18403611 TI - Nucleophosmin (NPM1) mutations in acute myeloid leukemia: an ongoing (cytoplasmic) tale of dueling mutations and duality of molecular genetic testing methodologies. PMID- 18403612 TI - High-throughput homogeneous mass cleave assay technology for the diagnosis of autosomal recessive Parkinson's disease. AB - To date, 11 gene loci that contribute to familial Parkinson's disease (PD) are known. Of these, mutations in six genes have been identified, allowing genetic testing and more accurate phenotypic characterization of genetically defined disease subtypes. In particular, mutations in Parkin, DJ-1, and Pink1 genes are associated with autosomal recessive PD and may also play a major role in early onset PD (EOPD). However, genetic testing for sequence alterations in these genes remains laborious. Therefore, our aim was to develop a flexible, rapid, high throughput screening procedure using matrix-assisted laser desorption ionization/time of flight technology and homogeneous mass cleave assays. Using this novel approach, we screened all 27 coding exons of the Parkin, DJ-1, and Pink1 genes in 31 patients with EOPD, a total of 367,195 nucleotides. Four positive controls with known autosomal recessive PD mutations that had previously been screened by denaturing high performance liquid chromatography in combination with sequencing were also tested. All known alterations were detected by matrix assisted laser desorption ionization/time of flight mass spectrometer, as well as additional polymorphisms in formerly unscreened regions. Overall, two previously described mutations in three patients with EOPD, 27 known polymorphisms with 386 occurrences, and eight unknown variants with 21 occurrences were detected. In total, we identified 410 sequence alterations in 31 patients with EOPD. In conclusion, this is the first study using matrix-assisted laser desorption ionization/time of flight mass spectrometry and homogeneous mass cleave assay for high-throughput mutation screening. PMID- 18403613 TI - An allele-specific rt-PCR assay to detect type A mutation of the nucleophosmin-1 gene in acute myeloid leukemia. AB - Nucleophosmin-1 (NPM1) mutations represent the most frequent gene alteration in acute myeloid leukemia (AML). The most common NPM1 mutation type, accounting for 75 to 80% of cases, is referred to as mutation A (NPM1-mutA). These NPM1 alterations have been shown to possess prognostic significance because they appear to identify patients who will benefit from chemotherapy. Given the high prevalence and stability of these mutations over the course of disease, NPM1 mutations may serve as ideal targets for minimal residual disease (MRD) assessment in AML. Current detection methods are costly, require sophisticated equipment, and are often not sufficiently sensitive. We report here an allele specific (ASO)-RT-PCR assay that enables rapid and sensitive detection of NPM1 mutA. A semi-nested ASO-PCR method was also designed to increase the sensitivity of our assay for the monitoring of MRD. We analyzed bone marrow cells collected from 52 patients with AML at presentation. NPM1-mutA was detected in leukemic cells from 21 patients. Assay specificity was confirmed by capillary electrophoresis and DNA sequencing. ASO-RT-PCR and semi-nested ASO-PCR assays could detect NPM1-mutA with sensitivities of 10(-2) and 10(-5), respectively. Results obtained here verify that our ASO-RT-PCR assay is a specific and sensitive method for the routine screening of NPM1-mutA, as well as for MRD monitoring of AML patients with this alteration. This method is convenient and easily applicable in countries with limited resources and no access to real-time quantitative PCR-based technologies. PMID- 18403614 TI - A single nucleotide variant in the FMR1 CGG repeat results in a "Pseudodeletion" and is not associated with the fragile X syndrome phenotype. AB - The molecular diagnosis of fragile X syndrome relies on the detection of the pathogenic CGG repeat expansion in the FMR1 gene. Deletions and point mutations have occasionally been reported. Rare polymorphisms might mimic a deletion by Southern blot analysis, leading to false-positive results. We describe a novel rare nucleotide substitution within the CGG repeat. The proband was a woman with a positive family history of mental retardation. Southern blot analysis showed an additional band consistent with a deletion in the region detected by the StB12.3 probe. Sequencing of this region revealed a G>C transversion that interrupts the CGG repeat and introduces an EagI site. The same variant was observed in both the healthy son and father of the proband, supporting the hypothesis that the nucleotide substitution is a silent polymorphism, the frequency of which we estimated to be less than 1% in the general population. These findings argue for a pathogenic role of nucleotide variants within the CGG repeat and suggest possible consequences of unexpected findings in the molecular diagnostics of fragile X syndrome. Thus, although the sequence context of a single nucleotide substitution may not predict possible effects on mRNA or protein function, a specific change in the higher order structures of DNA or mRNA may be functionally relevant in the pathological phenotype. PMID- 18403615 TI - Glitazones in type 2 diabetes: an update. AB - The glitazones and their combination products are licensed for type 2 diabetes mellitus and now account for over 50% of NHS expenditure on oral hypoglycaemic drugs. In 2001, for pioglitazone and rosiglitazone (the two glitazones licensed in the UK), we concluded that evidence (from short-term studies) supported their use in combination with metformin or a sulphonylurea in patients unsuited to one or other of these drugs, but not more practical glitazone use, such as monotherapy or as part of triple therapy. We also said that their long-term effects were not yet clear. Since then, both drugs have also been licensed for monotherapy, as part of triple therapy that includes metformin and a sulphonylurea and, for pioglitazone, use with insulin. Here we reassess glitazones in light of published efficacy data and safety concerns raised in recent years. PMID- 18403616 TI - Depression, antidepressants and heart disease. AB - Around 1 in 5 people with coronary artery disease has major depression and the prognosis in heart disease is worse in those who are also depressed. Many antidepressants have unwanted cardiovascular effects, so choosing between them may not be straightforward in patients with heart disease. Here, we consider whether any of the commonly used antidepressants are more or less likely to exacerbate heart disease and whether treatment for depression affects cardiac outcomes. PMID- 18403617 TI - Specific overexpression of IL-7 in the intestinal mucosa: the role in intestinal intraepithelial lymphocyte development. AB - IL-7 plays a crucial role in controlling T cell development and homeostasis. Since IL-7 may be derived from extraintestinal sources, and exogenous IL-7 broadly affects lymphoid populations, the actions of epithelial cell (EC)-derived IL-7 are not fully understood. The effect of intestinal specific expression of IL 7 on intestinal mucosal lymphocytes was investigated by using an IL-7 transgenic mouse model. We generated an intestinal EC-specific overexpressing IL-7 transgenic mouse model (IL-7(vill)) and compared their phenotype and function to wild-type C57BL/6J mice. EC-derived IL-7 overexpression was found to be exclusively in the small and large intestine. Numbers and subtypes of mucosal lymphocytes, including intraepithelial lymphocytes (IEL) and lamina propria lymphocytes (LPL), significantly changed in IL-7(vill) mice. From a functional standpoint, IEL proliferation also significantly increased in IL-7(vill) mice. IEL cytokine expression significantly changed in both T cell receptor (TCR) alphabeta(+) and TCR-gammadelta(+) IEL subpopulations, including a significant increase in IFN-gamma and TNF-alpha as well as an increase in keratinocyte growth factor expression. EC expression of CD103 (integrin alpha(E)beta(7)), the ligand of E-cadherin, markedly upregulated and may account for a mechanism of the massive expansion of IEL in transgenic mice. Systemic lymphoid populations did not change in transgenic mice. IL-7 overexpression by intestinal EC significantly affected IEL phenotype and function. These results offer insight into the role of IL-7 in IEL development and suggest a critical role of EC-derived expression of IL-7 in the phenotype and function of IEL. PMID- 18403618 TI - Salmonella enterica serovar Typhimurium modulates P-glycoprotein in the intestinal epithelium. AB - Studies over the last decade have shown that Salmonella enterica serovar Typhimurium (S. typhimurium) is able to preferentially locate to sites of tumor growth and modulate (shrink) the growth of many cancers. Given this unique association between S. typhimurium and cancer cells, the objective of this study was to investigate the capacity of this microorganism to modulate the plasma membrane multidrug resistance (MDR) protein P-glycoprotein (P-gp), an ATP-binding cassette transporter responsible for effluxing many cancer drugs. Using an in vitro model of S. typhimurium infection of polarized human cancer intestinal cell lines, we have found that this enteric pathogen functionally downregulates the efflux capabilities of P-gp. Specifically, we show that S. typhimurium infection of human intestinal cancer cells results in the enhanced intracellular accumulation of a number of P-gp substrates that corresponds to the posttranscriptional downregulation of P-gp expression. Furthermore, cells expressing small interfering RNAs against MDR1, the gene encoding P-gp, were significantly more susceptible to the cytotoxic effects of bacterial infection. This result is consistent with our observation that S. typhimurium was significantly less able to invade cells overexpressing MDR1. Taken together, these results reveal a novel role for P-gp in the maintenance of homeostasis in the gastrointestinal tract in regard to bacterial infection. Thus the regulation of P-gp by S. typhimurium has important implications not only for the development of new cancer therapeutics aimed at reversing drug resistance but also in the understanding of how microbes have evolved diverse strategies to interact with their host. PMID- 18403619 TI - Differences in intragastric pH in diabetic vs. idiopathic gastroparesis: relation to degree of gastric retention. AB - Evidence suggests that distinct mechanisms underlie diabetic and idiopathic gastroparesis. Differences in gastric acid in gastroparesis of different etiologies and varying degrees of gastric stasis are uninvestigated. We tested the hypotheses that 1) gastric pH profiles show differential alteration in diabetic vs. idiopathic gastroparesis and 2) abnormal pH profiles relate to the severity of gastric stasis. Sixty-four healthy control subjects and 44 gastroparesis patients (20 diabetic, 24 idiopathic) swallowed wireless transmitting capsules and then consumed (99m)Tc-sulfur colloid-labeled meals for gastric scintigraphy. Gastric pH from the capsule was recorded every 5 s. Basal pH was higher in diabetic (3.64 +/- 0.41) vs. control subjects (1.90 +/- 0.18) and idiopathic subjects (2.41 +/- 0.42; P < 0.05). Meals evoked initial pH increases that were greater in diabetic (4.98 +/- 0.32) than idiopathic patients (3.89 +/- 0.39; P = 0.03) but not control subjects (4.48 +/- 0.14). pH nadirs prior to gastric capsule evacuation were higher in diabetic patients (1.50 +/- 0.23) than control subjects (0.58 +/- 0.11; P = 0.003). Four-hour gastric retention was similar in diabetic (18.3 +/- 0.5%) and idiopathic (19.4 +/- 0.5%) patients but higher than control subjects (2.2 +/- 0.5%; P < 0.001). Compared with control subjects, those with moderate-severe stasis (>20% retention at 4 h) had higher basal (3.91 +/- 0.55) and nadir pH (2.23 +/- 0.42) values (P < 0.05). In subgroup analyses, both diabetic and idiopathic patients with moderate-severe gastroparesis exhibited increased pH parameters vs. those with mild gastroparesis. In conclusion, diabetic patients with gastroparesis exhibit reduced gastric acid, an effect more pronounced in those with severely delayed gastric emptying. Idiopathic gastroparetic subjects exhibit nearly normal acid profiles, although those with severely delayed emptying show reduced acid vs. those with mild delays. Thus both etiology and degree of gastric stasis determine gastric acidity in gastroparesis. PMID- 18403620 TI - Characteristics and outcomes of patients with chronic myeloid leukemia and T315I mutation following failure of imatinib mesylate therapy. AB - Chronic myeloid leukemia (CML) with T315I mutation has been reported to have poor prognosis. We analyzed 27 patients with T315I, including 20 who developed T315I after imatinib failure (representing 11% of 186 patients with imatinib failure), and 7 of 23 who developed new mutations after second tyrosine kinase inhibitor (TKI). Median follow-up from mutation detection was 18 months. At the time of T315I detection, 10 were in chronic phase (CP), 9 in accelerated phase, and 8 in blast phase. Except for the lack of response to second TKIs (P = .002), there was no difference in patient characteristics and outcome between patients with T315I and those with other or no mutations. Patients in CP had a 2-year survival rate of 87%. Although the T315I mutation is resistant to currently available TKIs, survival of patients with T315I remains mostly dependent on the stage of the disease, with many CP patients having an indolent course. PMID- 18403621 TI - Quantitative assessment of colorectal cancer tumor vascular parameters by using perfusion CT: influence of tumor region of interest. AB - PURPOSE: To prospectively determine whether position and size of tumor region of interest (ROI) influence estimates of colorectal cancer vascular parameters at computed tomography (CT). MATERIALS AND METHODS: After institutional review board approval and informed consent, 25 men and 22 women (mean age, 65.8 years) with colorectal adenocarcinoma underwent 65-second CT perfusion study. Blood volume, blood flow, and permeability-surface area product were determined for 40- or 120 mm(2) circular ROIs placed at the tumor edge and center and around (outlining) visible tumor. ROI analysis was repeated by two observers in different subsets of patients to assess intra- and interobserver variation. Measurements were compared by using analysis of variance; a difference with P = .002 was significant. RESULTS: Blood volume, blood flow, and permeability-surface area product measurements were substantially higher at the edge than at the center for both 40 and 120-mm(2) ROIs. For 40-mm(2) ROI, means of the three measurements were 6.9 mL/100 g (standard deviation [SD], 1.4), 108.7 mL/100 g per minute (SD, 39.2), and 16.9 mL/100 g per minute (SD, 4.2), respectively, at the edge versus 5.1 mL/100 g (SD, 1.5), 56.3 mL/100 g per minute (SD, 33.1), and 13.9 mL/100 g per minute (SD, 4.6), respectively, at the center. For 120-mm(2) ROI, means of the three measurements were 6.6 mL/100 g (SD, 1.3), 96.7 mL/100 g per minute (SD, 42.5), and 16.3 mL/100 g per minute (SD, 5.6), respectively, at the edge versus 5.1 mL/100 g (SD, 1.4), 58.3 mL/100 g per minute (SD, 32.5), and 13.4 mL/100 g per minute (SD, 4.3) at the center (P < .0001). Measurements varied substantially depending on the ROI size; values for the ROI for outlined tumor were intermediate between those at the tumor edge and center. Inter- and intraobserver agreement was poor for both 40- and 120-mm(2) ROIs. CONCLUSION: Position and size of tumor ROI and observer variation substantially influence ultimate perfusion values. ROI for outlined entire tumor is more reliable for perfusion measurements and more appropriate clinically than use of arbitrarily determined smaller ROIs. PMID- 18403622 TI - Induced hepatic fibrosis in rats: hepatic steatosis, macromolecule content, perfusion parameters, and their correlations--preliminary MR imaging in rats. AB - PURPOSE: To prospectively evaluate magnetic resonance (MR) imaging for the characterization of liver fibrosis by estimating fat and extracellular matrix content and hepatic perfusion parameters in CCl(4)-treated rats. MATERIALS AND METHODS: The animal research protocol was approved by the Institutional Animal Care and Use Committee. Fifty-two rats (38 treated, 14 control) were included. A CCl(4) mixture was injected three times per week for 2-16 weeks. Fat-to-water ratios (FWRs) were calculated. Images were obtained with 12 saturation offset frequencies; magnetization transfer ratios (MTRs) were calculated. Distribution volume (DV), mean transit time (MTT), and portal fraction (PF) of blood inflow were calculated. For pairwise group comparisons, an unequal two-tailed Student t test was used. For pairwise correlations between variables, Pearson correlation coefficients were calculated. For multiple pairwise comparisons, Bonferroni correction was performed by adjusting the significance level (alpha). RESULTS: FWR and DV were correlated with CCl(4) treatment duration from 0 through 8 weeks (r = 0.658, P < .001 and r = -0.664, P < .001, respectively; alpha = .010). PF and MTT were correlated with CCl(4) treatment duration from 0 through 16 weeks (r = -0.483, P = .002 and r = 0.414, P = .008, respectively; alpha = .010). DV was inversely correlated with FWR over the same period (r = -0.581, P < .001; alpha = .007). Fibrotic rats without cirrhosis had a higher FWR and lower DV and PF (P < .001, P < .001, and P = .004, respectively; alpha = .017) than control rats, and lower MTR, DV, and MTT (P = .014, .001, and .010, respectively; alpha = .017) than cirrhotic rats. Cirrhotic rats had a higher FWR and a lower PF (P < .001, alpha = .017) than control rats. CONCLUSION: Magnetization transfer contrast is not a specific indicator of increased fibrosis in diseased liver; steatosis may influence some perfusion parameters. PMID- 18403623 TI - Renal oxygenation changes during acute unilateral ureteral obstruction: assessment with blood oxygen level-dependent mr imaging--initial experience. AB - PURPOSE: To prospectively determine if changes in intrarenal oxygenation during acute unilateral ureteral obstruction can be depicted with blood oxygen level dependent (BOLD) magnetic resonance (MR) imaging. MATERIALS AND METHODS: The study was approved by the local ethics committee, and written informed consent was obtained from all patients. BOLD MR imaging was performed in 10 male patients (mean age, 45 years +/- 17 [standard deviation]; range, 20-73 years) with a distal unilateral ureteral calculus and in 10 healthy age-matched male volunteers to estimate R2*, which is inversely related to tissue Po(2). R2* values were determined in the cortex and medulla of the obstructed and the contralateral nonobstructed kidneys. To reduce external effects on R2*, the R2* ratio between the medulla and cortex was also analyzed. Statistical analysis was performed with nonparametric rank tests. P < .05 was considered to indicate a significant difference. RESULTS: All patients had significantly lower medullary and cortical R2* values in the obstructed kidney (median R2* in medulla, 10.9 sec(-1) [range, 9.1-14.3 sec(-1)]; median R2* in cortex, 10.4 sec(-1) [range, 9.7-11.3 sec(-1)]) than in the nonobstructed kidney (median R2* in medulla, 17.2 sec(-1) [range, 14.6-23.2 sec(-1)], P = .005; median R2* in cortex, 11.7 sec(-1) [range, 11.0 14.0 sec(-1)], P = .005); values in the obstructed kidneys were also significantly lower than values in the kidneys of healthy control subjects (median R2* in medulla, 16.1 sec(-1) [range, 13.9-18.1 sec(-1)], P < .001; median R2* in cortex, 11.6 sec(-1) [range, 10.5-12.9 sec(-1)], P < .001). R2* ratios in the obstructed kidneys (median, 1.06; range, 0.85-1.27) were significantly lower than those in the nonobstructed kidneys (median, 1.49; range, 1.26-1.71; P = .005) and those in the kidneys of healthy control subjects (median, 1.38; range, 1.23-1.47; P < .001). In contrast, R2* ratios in the nonobstructed kidneys of patients were significantly higher than those in kidneys of healthy control subjects (P = .01). CONCLUSION: Increased oxygen content in the renal cortex and medulla occurs with acute unilateral ureteral obstruction, suggesting reduced function of the affected kidney. PMID- 18403624 TI - Benign and malignant thyroid nodules: US differentiation--multicenter retrospective study. AB - PURPOSE: To retrospectively evaluate the diagnostic accuracy of ultrasonographic (US) criteria for the depiction of benign and malignant thyroid nodules by using tissue diagnosis as the reference standard. MATERIALS AND METHODS: This study had institutional review board approval, and informed consent was waived. From January 2003 through June 2003, 8024 consecutive patients had undergone thyroid US at nine affiliated hospitals. A total of 831 patients (716 women, 115 men; mean age, 49.5 years +/- 13.8 [standard deviation]) with 849 nodules (360 malignant, 489 benign) that were diagnosed at surgery or biopsy were included in this study. Three radiologists retrospectively evaluated the following characteristics on US images: nodule size, presence of spongiform appearance, shape, margin, echotexture, echogenicity, and presence of microcalcification, macrocalcification, or rim calcification. A chi(2) test and multiple regression analysis were performed. Sensitivity, specificity, and positive and negative predictive values were obtained. RESULTS: Statistically significant (P < .05) findings of malignancy were a taller-than-wide shape (sensitivity, 40.0%; specificity, 91.4%), a spiculated margin (sensitivity, 48.3%; specificity, 91.8%), marked hypoechogenicity (sensitivity, 41.4%; specificity, 92.2%), microcalcification (sensitivity, 44.2%; specificity, 90.8%), and macrocalcification (sensitivity, 9.7%; specificity, 96.1%). The US findings for benign nodules were isoechogenicity (sensitivity, 56.6%; specificity, 88.1%; P < .001) and a spongiform appearance (sensitivity, 10.4%; specificity, 99.7%; P < .001). The presence of at least one malignant US finding had a sensitivity of 83.3%, a specificity of 74.0%, and a diagnostic accuracy of 78.0%. For thyroid nodules with a diameter of 1 cm or less, the sensitivity of microcalcifications was lower than that in larger nodules (36.6% vs 51.4%, P < .05). CONCLUSION: Shape, margin, echogenicity, and presence of calcification are helpful criteria for the discrimination of malignant from benign nodules; the diagnostic accuracy of US criteria is dependent on tumor size. PMID- 18403625 TI - Redundant publication in the journal Radiology. AB - PURPOSE: To retrospectively quantify the incidence of redundant publication in the journal Radiology and to compare the present study findings with those published for other journals and medical specialties. MATERIALS AND METHODS: Two readers estimated the incidence of the redundant publication of original articles in Radiology in the year 2001. Original research articles published in 2001 were analyzed by searching MEDLINE on the PubMed server to identify articles that may have represented a duplication of the original Radiology article. MEDLINE was searched between January 1999 and December 2003 by using the surname and initial(s) of the first author. Potentially redundant articles were identified after similarities in titles and abstracts were analyzed. The full versions of all potentially redundant articles and of the corresponding index articles were then retrieved from the library. The potentially redundant article was then compared with the index article. Criteria for redundant publication were as follows: Compared with the index article, the potentially duplicate article had (a) a similar hypothesis, (b) a similar number of subjects, (c) similar results, (d) at least one author in common, and (e) no or little new information. RESULTS: In 2001, 362 original research articles were published in Radiology. Two instances of redundant publication were found among these articles, and both were considered to be partially redundant publications due to series expansions (ie, increased numbers of study subjects) of 50% and 52%. CONCLUSION: Redundant publication appears to be less frequent in Radiology than in the other journals and specialties for which redundant publication information has been reported. PMID- 18403627 TI - Recurrent lower-limb varicose veins: effect of direct contrast-enhanced three dimensional MR venographic findings on diagnostic thinking and therapeutic decisions. AB - PURPOSE: To assess the effect of direct three-dimensional (3D) magnetic resonance (MR) venographic findings on diagnostic thinking and therapeutic decisions in patients with complex recurrent varicose vein anatomy who were being evaluated for surgical treatment. MATERIALS AND METHODS: The study was approved by the Institutional Review Board; informed consent was obtained from patients. MR venography was performed before surgery in 22 legs of 14 patients (seven women: mean age, 53 years; seven men: mean age, 59 years) thought to have recurrent varicose veins. Two radiologists assessed image quality and evaluated sites and sources of varicose veins on MR venograms. One vascular surgeon completed a questionnaire before and after MR venography and noted diagnosis and therapeutic decisions. Diagnoses at MR venography were compared with surgical results in 19 legs that underwent surgery. Differences between diagnosed and treated varicose veins per leg before and after MR venography were analyzed with logistic regression for survey data. kappa Values were calculated to illustrate interobserver agreement for grading image quality of venous segments and for diagnosing recurrent varicose veins. RESULTS: Mean graded image quality of the deep venous system and the recurrent varicose veins was good or excellent in 89% of segments. There was good agreement between readers regarding grading of image quality of venous segments (kappa = 0.80). After MR venography, diagnosis of the sites and sources of recurrent varicose veins changed in 17 of 22 legs of nine of 14 patients. In one of 14 patients, the preoperative diagnosis of recurrent varicose veins was withdrawn. A change in treatment plan occurred in 17 of 22 legs after MR venography. The number of diagnosed and treated sources of reflux increased significantly after MR venography. MR venographic diagnoses were confirmed at surgery in all 19 legs. CONCLUSION: MR venographic results have a substantial effect on diagnostic thinking and therapeutic decisions when recurrent lower-limb varicose veins are suspected. PMID- 18403626 TI - Postinfarction myocardial scarring in mice: molecular MR imaging with use of a collagen-targeting contrast agent. AB - PURPOSE: To prospectively evaluate a gadolinium-based collagen-targeting contrast agent, EP-3533, for in vivo magnetic resonance (MR) imaging of myocardial fibrosis in a mouse model of healed myocardial infarction (MI). MATERIALS AND METHODS: All procedures were performed in accordance with protocols approved by the animal care and use committee. MI was induced in eight mice by means of occlusion of the left anterior descending coronary artery followed by reperfusion. Four MR examinations were performed in each animal: one examination before, one examination 1 day after, and two examinations 6 weeks after the MI. For the latter two examinations, electrocardiographically gated inversion recovery gradient-echo MR images were acquired before and serially (every 5 minutes) after the intravenous injection of either gadopentetate dimeglumine or EP-3533. The image enhancement kinetic properties of the postinfarction scar, normal myocardium, and blood were compared. RESULTS: Dynamic T1-weighted MR imaging revealed the washout time constants for EP-3533 to be significantly longer than those for gadopentetate dimeglumine in regions of postinfarction scarring (mean, 194.8 minutes +/-116.8 [standard deviation] vs 25.5 minutes +/- 4.2; P < .05) and in normal myocardium (mean, 45.4 minutes +/- 16.7 vs 25.1 minutes +/- 9.7; P < .05). Findings on postmortem histologic sections stained for collagen correlated well with EP-3533-enhanced areas seen on inversion-recovery MR images. Fifty minutes after EP-3533 injection, the postinfarction scar tissue samples, as compared with the normal myocardium, had a twofold higher concentration of gadolinium. CONCLUSION: Use of the gadolinium-based collagen targeting contrast agent, EP-3533, enabled in vivo molecular MR imaging of fibrosis in a mouse model of healed postinfarction myocardial scarring. PMID- 18403628 TI - Direct MR arthrography at 1.5 and 3.0 T: signal dependence on gadolinium and iodine concentrations--phantom study. AB - PURPOSE: To prospectively quantify in vitro the influence of gadopentetate dimeglumine and ioversol on the magnetic resonance (MR) imaging signal observed with a variety of musculoskeletal pulse sequences to predict optimum gadolinium concentrations for direct MR arthrography at 1.5 and 3.0 T. MATERIALS AND METHODS: In an in vitro study, T1 and T2 relaxation times of three dilution series of gadopentetate dimeglumine (concentration, 0-20.0 mmol gadolinium per liter) at ioversol concentrations with iodine concentration of 0, 236.4, and 1182 mmol iodine per liter (corresponding to 0, 30, and 150 mg of iodine per milliliter) were measured at 1.5 and 3.0 T. The relaxation rate dependence on concentrations of gadolinium and iodine was analytically modeled, and continuous profiles of signal versus gadolinium concentration were calculated for 10 pulse sequences used in current musculoskeletal imaging. After fitting to experimental discrete profiles, maximum signal-to-noise ratio (SNR), gadolinium concentration with maximum SNR, and range of gadolinium concentration with 90% of maximum SNR were derived. The overall influence of field strength and iodine concentration on these parameters was assessed by using t tests. The deviation of simulated from experimental signal-response profiles was assessed with the autocorrelation of the residuals. RESULTS: The model reproduced relaxation rates of 0.37-38.24 sec( 1), with a mean error of 4.5%. Calculated SNR profiles matched the discrete experimental profiles, with autocorrelation of the residuals divided by the mean of less than 5.0. Admixture of ioversol consistently reduced T1 and T2, narrowed optimum gadolinium concentration ranges (P = .004-.006), and reduced maximum SNR (P < .001 to not significant). Optimum gadolinium concentration was 0.7-3.4 mmol/L at both field strengths. At 3.0 T, maximum SNR was up to 75% higher than at 1.5 T. CONCLUSION: Admixture of ioversol to gadopentetate dimeglumine solutions results in a consistent additional relaxation enhancement, which can be analytically modeled to allow a near-quantitative a priori optimized match of contrast media concentrations and imaging protocol for a broad variety of pulse sequences. PMID- 18403629 TI - Diagnostic performance of 16- and 64-section spiral CT for coronary artery bypass graft assessment: meta-analysis. AB - PURPOSE: To perform a meta-analysis to evaluate the accuracy of 16- and 64 section spiral computed tomography (CT) to help assess coronary artery bypass grafts (CABGs). MATERIALS AND METHODS: The MEDLINE, Cochrane library, and BioMed Central databases were searched for relevant original articles published up to May 2007. Major criteria for article inclusion were that it (a) used multisection CT as a diagnostic test for the assessment of significant lesions (occlusion or >50% stenosis) of CABG, (b) used a 16- or 64-section scanner, and (c) used coronary angiography as the reference standard. After data extraction, the analysis was performed according to a random-effects model. Between-study statistical heterogeneity was also assessed by using the Cochran Q chi(2) test. RESULTS: Of 158 screened articles, 15 fulfilled all inclusion criteria. Graft assessability (including distal anastomosis) ranged from 78%-100% among all included studies (mean, 92.4%; 90% with 16- and 96% with 64-section CT; P < .001). Statistical heterogeneity was observed for specificity and positive likelihood ratio (LR), justifying the use of the random-effects model. The analysis, pooled from 15 studies (723 patients, 2023 CABGs), provided the following results for the assessment of graft obstruction (occlusion and >50% stenosis): sensitivity, 97.6% (95% confidence interval [CI]: 96%, 98.6%); specificity, 96.7% (95% CI: 95.6%, 97.5%); positive predictive value, 92.7% (95% CI: 90.5%, 94.6%); negative predictive value, 98.9% (95% CI: 98.2%, 99.4%); positive LR, 23.42 (95% CI: 13.69, 40.07); negative LR, 0.045 (95% CI: 0.028, 0.071); and diagnostic odds ratio, 780.32 (95% CI: 379.12, 1606.1). CONCLUSION: Multisection CT provided high accuracy for the evaluation of CABG obstruction in assessable conduits, and might be used as a noninvasive tool for the evaluation of suspected graft dysfunction in patients who are at high risk for complications from coronary angiography. PMID- 18403630 TI - Integration of reporter transgenes into Schistosoma mansoni chromosomes mediated by pseudotyped murine leukemia virus. AB - The recent release of draft genome sequences of two of the major human schistosomes has underscored the pressing need to develop functional genomics approaches for these significant pathogens. The sequence information also makes feasible genome-scale investigation of transgene integration into schistosome chromosomes. Retrovirus-mediated transduction offers a means to establish transgenic lines of schistosomes, to elucidate schistosome gene function and expression, and to advance functional genomics approaches for these parasites. We investigated the utility of the Moloney murine leukemia retrovirus (MLV) pseudotyped with vesicular stomatitis virus glycoprotein (VSVG) for the transduction of Schistosoma mansoni and delivery of reporter transgenes into schistosome chromosomes. Schistosomula were exposed to virions of VSVG pseudotyped MLV, after which genomic DNA was extracted from the transduced schistosomes. Southern hybridization analysis indicated the presence of proviral MLV retrovirus in the transduced schistosomes. Fragments of the MLV transgene and flanking schistosome sequences recovered using an anchored PCR-based approach demonstrated definitively that somatic transgenesis of schistosome chromosomes had taken place and, moreover, revealed widespread retrovirus integration into schistosome chromosomes. More specifically, MLV transgenes had inserted in the vicinity of genes encoding immunophilin, zinc finger protein Sma-Zic, and others, as well as near the endogenous schistosome retrotransposons, the fugitive and SR1. Proviral integration of the MLV transgene appeared to exhibit primary sequence site specificity, targeting a gGATcc-like motif. Reporter luciferase transgene activity driven by the schistosome actin gene promoter was expressed in the tissues of transduced schistosomula and adult schistosomes. Luciferase activity appeared to be developmentally expressed in schistosomula with increased activity observed after 1 to 2 wk in culture. These findings indicate the utility of VSVG-pseudotyped MLV for transgenesis of S. mansoni, herald a tractable pathway forward toward germline transgenesis and functional genomics of parasitic helminths, and provide the basis for comparative molecular pathogenesis studies of chromosomal lesions arising from retroviral integration into human compared with schistosome chromosomes. PMID- 18403632 TI - DNA damage responses: mechanisms and roles in human disease: 2007 G.H.A. Clowes Memorial Award Lecture. AB - Significant progress has been made in recent years in elucidating the molecular controls of cellular responses to DNA damage in mammalian cells. Much of our understanding of the mechanisms involved in cellular DNA damage response pathways has come from studies of human cancer susceptibility syndromes that are altered in DNA damage responses. Ataxia-telangiectasia mutated (ATM), the gene mutated in the disorder ataxia-telangiectasia, codes for a protein kinase that is a central mediator of responses to DNA double-strand breaks (DSB) in cells. Once activated, ATM phosphorylates numerous substrates in the cell that modulate the response of the cell to the DNA damage. We recently developed a novel system to create DNA DSBs at defined endogenous sites in the human genome and used this system to detect protein recruitment and loss at and around these breaks by chromatin immunoprecipitation. Results from this system showed the functional importance of ATM kinase activity and phosphorylation in the response to DSBs and supported a model in which ordered chromatin structure changes that occur after DNA breakage and that depend on functional NBS1 and ATM facilitate DNA DSB repair. Insights about these pathways provide us with opportunities to develop new approaches to benefit patients. Examples and opportunities for developing inhibitors that act as sensitizers to chemotherapy or radiation therapy or activators that could improve responses to cellular stresses, such as oxidative damage, are discussed. Relevant to the latter, we have shown benefits of an ATM activator in disease settings ranging from metabolic syndrome to cancer prevention. PMID- 18403631 TI - Calcium-permeable AMPA receptors containing Q/R-unedited GluR2 direct human neural progenitor cell differentiation to neurons. AB - We identify calcium-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors on human neural progenitor cells (NPCs) and present a physiological role in neurogenesis. RNA editing of the GluR2 subunit at the Q/R site is responsible for making most AMPA receptors impermeable to calcium. Because a single-point mutation could eliminate the need for editing at the Q/R site and Q/R-unedited GluR2 exists during embryogenesis, the Q/R-unedited GluR2 subunit presumably has some important actions early in development. Using calcium imaging, we found that NPCs contain calcium-permeable AMPA receptors, whereas NPCs differentiated to neurons and astrocytes express calcium-impermeable AMPA receptors. We utilized reverse-transcription polymerase chain reaction and BbvI digestion to demonstrate that NPCs contain Q/R-unedited GluR2, and differentiated cells contain Q/R-edited GluR2 subunits. This is consistent with the observation that the nuclear enzyme responsible for Q/R-editing, adenosine deaminase (ADAR2), is increased during differentiation. Activation of calcium-permeable AMPA receptors induces NPCs to differentiate to the neuronal lineage and increases dendritic arbor formation in NPCs differentiated to neurons. AMPA-induced differentiation of NPCs to neurons is abrogated by overexpression of ADAR2 in NPCs. This elucidates the role of AMPA receptors as inductors of neurogenesis and provides a possible explanation for why the Q/R editing process exists. PMID- 18403633 TI - Vascular endothelial growth factor stimulates organ-specific host matrix metalloproteinase-9 expression and ovarian cancer invasion. AB - Vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMP) regulate each other, contributing to tumor progression. We have previously reported that MMP9 induces the release of tumor VEGF, promoting ascites formation in human ovarian carcinoma xenografts. The aim of this study was to investigate whether tumor-derived VEGF regulated the expression of gelatinase by the stroma, influencing the invasive properties of ovarian tumors. Tumor variants derived from 1A9 human ovarian carcinoma, stably expressing VEGF(121) in the sense (1A9 VS-1) and antisense orientations (1A9-VAS-3), were used. In vivo, zymographic analysis of tumors from 1A9-VS-1 implanted in the peritoneal cavity of nude mice showed higher levels of gelatinases, particularly murine MMP9, indicating that VEGF stimulates host expression of the matrix-degrading enzyme. Murine MMP9 expression was also high in the ovaries of mice bearing 1A9-VS-1 tumors. The effect on host MMP9 activity was organ-specific. The levels of host pro-MMP9 in ovaries correlated with the plasma levels of tumor VEGF and with the selective invasion of the ovaries. Induction of host MMP9 expression in tumors and ovaries was independent of the site of tumor growth as it was seen in mice carrying both intraperitoneal and subcutaneous tumors. The anti-VEGF antibody bevacizumab (Avastin) inhibited MMP9 expression and tumor invasion in the ovaries of mice bearing 1A9-VS-1 tumors. These findings point to a complex cross-talk between VEGF and MMPs in the progression of ovarian tumor and suggest the possibility of using VEGF inhibitors to affect MMP-dependent tumor invasion. PMID- 18403634 TI - Arachidonic acid-induced Ca2+ entry is involved in early steps of tumor angiogenesis. AB - Growth factor-induced intracellular calcium signals in endothelial cells regulate cytosolic and nuclear events involved in the angiogenic process. Among the intracellular messengers released after proangiogenic stimulation, arachidonic acid (AA) plays a key role and its effects are strictly related to calcium homeostasis and cell proliferation. Here, we studied AA-induced intracellular calcium signals in endothelial cells derived from human breast carcinomas (B TEC). AA promotes B-TEC proliferation and organization of vessel-like structures in vitro. The effect is directly mediated by the fatty acid without a significant contribution of its metabolites. AA induces Ca(2+)(i) signals in the entire capillary-like structure during the early phases of tubulogenesis in vitro. No such responses are detectable in B-TECs organized in more structured tubules. In B-TECs growing in monolayer, AA induces two different signals: a Ca(2+)(i) increase due to Ca(2+) entry and an inhibition of store-dependent Ca(2+) entry induced by thapsigargin or ATP. An inhibitor of Ca(2+) entry and angiogenesis, carboxyamidotriazole, significantly and specifically decreases AA-induced B-TEC tubulogenesis, as well as AA-induced Ca(2+) signals in B-TECs. We conclude that (a) AA-activated Ca(2+) entry is associated with the progression through the early phases of angiogenesis, mainly involving proliferation and tubulogenesis, and it is down-regulated during the reorganization of tumor-derived endothelial cells in capillary-like structures; and (b) inhibition of AA-induced Ca(2+) entry may contribute to the antiangiogenic action of carboxyamidotriazole. PMID- 18403635 TI - MUC1 initiates Src-CrkL-Rac1/Cdc42-mediated actin cytoskeletal protrusive motility after ligating intercellular adhesion molecule-1. AB - MUC1, a transmembrane glycoprotein of the mucin family, when aberrantly expressed on breast cancer cells is correlated with increased lymph node metastases. We have previously shown that MUC1 binds intercellular adhesion molecule-1 (ICAM-1) on surrounding accessory cells and facilitates transendothelial migration of MUC1 bearing cells. Nevertheless, the underlying molecular mechanism is still obscure. In the present study, we used a novel assay of actin cytoskeletal reorganization to show that by ligating ICAM-1, MUC1 triggers Rac1- and Cdc42-dependent actin cytoskeletal protrusive activity preferentially at the heterotypic cell-cell contact sites. Further, we show that these MUC1/ICAM-1 interaction-initiated lamellipodial and filopodial protrusions require Src family kinase and CT10 regulator of kinase like (CrkL) accompanied by the rapid formation of a Src-CrkL signaling complex at the MUC1 cytoplasmic domain. Through inhibition of Src kinase activity, we further revealed that Src is required for recruiting CrkL to the MUC1 cytoplasmic domain as well as mediating the observed actin cytoskeleton dynamics. These findings suggest a novel MUC1-Src-CrkL-Rac1/Cdc42 signaling cascade following ICAM-1 ligation, through which MUC1 regulates cytoskeletal reorganization and directed cell motility during cell migration. PMID- 18403636 TI - A different view on DNA amplifications indicates frequent, highly complex, and stable amplicons on 12q13-21 in glioma. AB - To further understand the biological significance of amplifications for glioma development and recurrencies, we characterized amplicon frequency and size in low grade glioma and amplicon stability in vivo in recurring glioblastoma. We developed a 12q13-21 amplicon-specific genomic microarray and a bioinformatics amplification prediction tool to analyze amplicon frequency, size, and maintenance in 40 glioma samples including 16 glioblastoma, 10 anaplastic astrocytoma, 7 astrocytoma WHO grade 2, and 7 pilocytic astrocytoma. Whereas previous studies reported two amplified subregions, we found a more complex situation with many amplified subregions. Analyzing 40 glioma, we found that all analyzed glioblastoma and the majority of pilocytic astrocytoma, grade 2 astrocytoma, and anaplastic astrocytoma showed at least one amplified subregion, indicating a much higher amplification frequency than previously suggested. Amplifications in low-grade glioma were smaller in size and displayed clearly different distribution patterns than amplifications in glioblastoma. One glioblastoma and its recurrencies revealed an amplified subregion of 5 Mb that was stable for 6 years. Expression analysis of the amplified region revealed 10 overexpressed genes (i.e., KUB3, CTDSP2, CDK4, OS-9, DCTN2, RAB3IP, FRS2, GAS41, MDM2, and RAP1B) that were consistently overexpressed in all cases that carried this amplification. Our data indicate that amplifications on 12q13-21 (a) are more frequent than previously thought and present in low-grade tumors and (b) are maintained as extended regions over long periods of time. PMID- 18403637 TI - The CpG island methylator phenotype correlates with long-range epigenetic silencing in colorectal cancer. AB - The CpG island methylator phenotype (CIMP), characterized by an exceptionally high frequency of methylation of discrete CpG islands, is observed in 18% to 25% of sporadic colorectal cancers. Another hypermethylation pattern found in colorectal cancers, termed long-range epigenetic silencing, is associated with DNA/histone methylation in three distinct gene clusters at chromosome 2q14.2, showing that DNA hypermethylation can span larger chromosomal domains and lead to the silencing of flanking, unmethylated genes. We investigated whether these two phenotypes are interrelated in colorectal cancers. The CIMP status of 148 sporadic colorectal cancers was determined by methylation-specific PCR. We determined the BRAF V600E mutation by mutant allele-specific PCR amplification. The methylation status of the MLH1 gene and of three CpG islands (EN1, SCTR, and INHBB), corresponding to three distinct clusters along 2q14.2, was determined by methylation-specific PCR. The average number of sites showing methylation in CIMP+ tumors was 2.21, compared with 1.22 for CIMP- individuals, and this difference was highly significant (P = 3.6 x 10(-8), Mann-Whitney test). Moreover, all CIMP+ tumors showed hypermethylation of at least one of these loci, in contrast to CIMP- tumors, where 18 (16%) samples remained unmethylated. The mean number of simultaneously hypermethylated CpG islands at 2q14.2 differs significantly between CIMP- and CIMP+ tumors, suggesting varying effects of domain silencing in this region. Given that the number of hypermethylated loci at 2q14.2 likely affects the range of silenced flanking genes, high frequency of simultaneous hypermethylation of three CpG islands (EN1, SCTR, and INHBB) may have potential influence on specific characteristics of CIMP+ colorectal cancers. PMID- 18403638 TI - Monochromosome transfer and microarray analysis identify a critical tumor suppressive region mapping to chromosome 13q14 and THSD1 in esophageal carcinoma. AB - Loss of chromosome 13q regions in esophageal squamous cell carcinoma (ESCC) is a frequent event. Monochromosome transfer approaches provide direct functional evidence for tumor suppression by chromosome 13 in SLMT-1, an ESCC cell line, and identify critical regions at 13q12.3, 13q14.11, and 13q14.3. Differential gene expression profiles of three tumor-suppressing microcell hybrids (MCH) and their tumorigenic parental SLMT-1 cell line were revealed by competitive hybridization using 19k cDNA oligonucleotide microarrays. Nine candidate 13q14 tumor-suppressor genes (TSG), including RB1, showed down-regulation in SLMT-1, compared with NE1, an immortalized normal esophageal epithelial cell line; their average gene expression was restored in MCHs compared with SLMT-1. Reverse transcription-PCR validated gene expression levels in MCHs and a panel of ESCC cell lines. Results suggest that the tumor-suppressing effect is not attributed to RB1, but instead likely involves thrombospondin type I domain-containing 1 (THSD1), a novel candidate TSG mapping to 13q14. Quantitative reverse transcription-PCR detected down-regulation of THSD1 expression in 100% of ESCC and other cancer cell lines. Mechanisms for THSD1 silencing in ESCC involved loss of heterozygosity and promoter hypermethylation, as analyzed by methylation-specific PCR and clonal bisulfite sequencing. Transfection of wild-type THSD1 into SLMT-1 resulted in significant reduction of colony-forming ability, hence providing functional evidence for its growth-suppressive activity. These findings suggest that THSD1 is a good candidate TSG. PMID- 18403639 TI - Localization of Fas/CD95 into the lipid rafts on down-modulation of the phosphatidylinositol 3-kinase signaling pathway. AB - Activation of the phosphatidylinositol 3-kinase (PI3K) signaling pathway is known to protect tumor cells from apoptosis and more specifically from the Fas-mediated apoptotic signal. The antitumoral agent edelfosine sensitizes leukemic cells to death by inducing the redistribution of the apoptotic receptor Fas into plasma membrane subdomains called lipid rafts. Herein, we show that inhibition of the PI3K signal by edelfosine triggers a Fas-mediated apoptotic signal independently of the Fas/FasL interaction. Furthermore, similarly to edelfosine, blockade of the PI3K activity, using specific inhibitors LY294002 and wortmannin, leads to the clustering of Fas whose supramolecular complex is colocalized within the lipid rafts. These findings indicate that the antitumoral agent edelfosine down modulates the PI3K signal to sensitize tumor cells to death through the redistribution of Fas into large platform of membrane rafts. PMID- 18403640 TI - Down-regulation of class II phosphoinositide 3-kinase alpha expression below a critical threshold induces apoptotic cell death. AB - Members of the phosphoinositide 3-kinase (PI3K) family collectively control multiple cellular responses, including proliferation, growth, chemotaxis, and survival. These diverse effects can partly be attributed to the broad range of downstream effectors being regulated by the products of these lipid kinases, the 3'-phosphoinositides. However, an additional layer of complexity is introduced by the existence of multiple PI3K enzyme isoforms. Much has been learned over the last years on the roles of the classes I and III PI3K members in cellular signaling, but little is known about the isoform-specific tasks done by the class II PI3Ks (C2alpha, beta, and gamma). In this study, we used quantitative reverse transcription-PCR and RNA interference in mammalian cells to gain further insight into the function of these lesser studied PI3K enzymes. We find that PI3K C2alpha, but not PI3K-C2beta, has an important role in controlling cell survival and by using a panel of RNA interference reagents, we were able to determine a critical threshold of PI3K-C2alpha mRNA levels, below which the apoptotic program is switched on, via the intrinsic cell death pathway. In addition, knockdown of PI3K-C2alpha to levels that by themselves do not induce apoptosis sensitize cells to the anticancer agent Taxol (paclitaxel). Lastly, we report that lowering the levels of PI3K-C2alpha in a number of cancer cell lines reduces their proliferation and cell viability, arguing that PI3K inhibitors targeting not only the class Ialpha isoform but also class IIalpha may contribute to an effective anticancer strategy. PMID- 18403641 TI - DUSP1 is controlled by p53 during the cellular response to oxidative stress. AB - p53 controls the cellular response to genotoxic stress through multiple mechanisms. We report here that p53 regulates DUSP1, a dual-specific threonine and tyrosine phosphatase with stringent substrate specificity for mitogen activated protein kinase (MAPK). DUSP1 is a potent inhibitor of MAPK activity through dephosphorylation of MAPK. In a colon cancer cell line containing inducible ectopic p53, DUSP1 protein level is significantly increased upon activation of p53, leading to cell death in response to nutritional stress. In mouse embryo fibroblast cells, DUSP1 protein abundance is greatly increased after oxidative stress in a p53-dependent manner and also when apoptosis is triggered. We show that p53 induces the activity of a human DUSP1 regulatory region. Furthermore, p53 can physically interact with the DUSP1 regulatory region in vivo, and p53 binds to a 10-bp perfect palindromic site in this DUSP1 regulatory region. We show that overexpression of DUSP1 or inhibition of MAPK activity significantly increases cellular susceptibility to oxidative damage. These findings indicate that p53 is a transcriptional regulator of DUSP1 in stress responses. Our results reveal a mechanism whereby p53 selectively regulates target genes and suggest a way in which subgroups of those target genes might be controlled independently. PMID- 18403642 TI - Prolactin does not require insulin-like growth factor intermediates but synergizes with insulin-like growth factor I in human breast cancer cells. AB - Insulin-like growth factor (IGF)-II is a required intermediate for prolactin induced up-regulation of cyclin D1 and proliferation in normal murine mammary epithelial cells in vivo and in vitro. However, we have recently shown that prolactin can rapidly induce cyclin D1 protein expression and subsequent proliferation in the MCF-7 human breast cancer cell line, suggesting that prolactin actions can be independent of IGFs in breast disease. Here, we investigate the relationship between these factors and show that prolactin up regulated transcript levels of both IGF-I and IGF-II, but only after increases in cyclin D1 protein were observed. Moreover, prolactin increased cyclin D1 in the presence of the IGF-I receptor neutralizing antibody alphaIR3. However, on cotreatment, IGF-I and prolactin elicited cooperative phosphorylation of extracellular signal-regulated kinases 1 and 2 and protein kinase B/AKT, but not signal transducer and activator of transcription 5. This interaction extended to increased activation of activating protein-1 enhancer elements, phosphorylation of glycogen synthase kinase 3beta, induction of cyclin D1, and ultimately, increased cell number. It also increased invasive behavior, which correlated with elevated matrix metalloproteinase-2 transcript levels. Interestingly, prolactin augmented phosphorylation at Tyr(1135) and Tyr(1136) of IGF-I receptor on cotreatment with IGF-I, although prolactin alone had no effect. Together, these data indicate that strong cooperative cross talk between prolactin and IGF-I augments biological processes associated with neoplastic progression, with implications for therapeutic strategies. PMID- 18403643 TI - Proteolytic processing of cut homeobox 1 by neutrophil elastase in the MV4;11 myeloid leukemia cell line. AB - Proteolytic processing by cathepsin L generates p110 Cut homeobox 1 (CUX1) at the end of the G(1) phase, whereas an alternative transcript encodes p75 CUX1. These short CUX1 isoforms were reported to be overexpressed in cancer cells, and transgenic mice overexpressing the p75 isoform were found to develop myeloproliferative disease-like myeloid leukemias. In the present study, we report that the neutrophil elastase can also generate a short CUX1 isoform in the MV4;11 acute myeloid leukemia cell line. Proteolytic processing was so efficient that the full-length CUX1 protein was detected only when cells were maintained in the presence of the specific elastase inhibitor III. In agreement with these findings, higher levels of the processed cyclin E isoforms were also detected in MV4;11 cells. Reappearance of full-length cyclin E and CUX1 could be induced upon the treatment of MV4;11 cells with the differentiation inducer phorbol 12 myristate 13-acetate or, unexpectedly, following overexpression of a short recombinant CUX1 protein. In both cases, the mechanism involved transcriptional repression of the neutrophil elastase gene. This result revealed a negative feedback loop whereby CUX1 shuts down the expression of the protease that cleaves it. Overall, the findings in MV4;11 and other cancer cells suggest that various mechanisms are used in cancer to favor the expression of short CUX1 isoforms. PMID- 18403644 TI - Hyperphosphorylated cortactin in cancer cells plays an inhibitory role in cell motility. AB - Cortactin is frequently overexpressed in cancer cells, and changes of the levels of its tyrosine phosphorylation have been observed in several cancer cells. However, how the expression level and phosphorylation state of cortactin would influence the ultimate cellular function of cancer cells is unknown. In this study, we analyzed the role of cortactin in gastric and breast cancer cell lines using RNA interference technique and found that knockdown of cortactin inhibited cell migration in a subset of gastric cancer cells with a lower level of its tyrosine phosphorylation, whereas it greatly enhanced cell migration and increased tyrosine phosphorylation of p130Cas in other subsets of cells with hyperphosphorylated cortactin. Consistent results were obtained when hyperphosphorylation of cortactin was induced in MCF7 breast cancer cells by expressing Fyn tyrosine kinase. Additionally, immunostaining analysis showed that knockdown of hyperphosphorylated cortactin resulted in the recruitment of p130Cas to focal adhesions. These results suggest that cortactin hyperphosphorylation suppresses cell migration possibly through the inhibition of membrane localization and tyrosine phosphorylation of p130Cas. PMID- 18403645 TI - Antiproliferative effects by Let-7 repression of high-mobility group A2 in uterine leiomyoma. AB - High-mobility group A2 (HMGA2) is commonly overexpressed in large leiomyomas. HMGA2 is an important regulator of cell growth, differentiation, apoptosis, and transformation. As a predicted target of Let-7 microRNAs (Let-7s), HMGA2 can be repressed by Let-7s in vitro. MicroRNA profiling analysis revealed that Let-7s were significantly dysregulated in uterine leiomyomas: high in small leiomyomas and lower in large leiomyomas. To evaluate whether Let-7 repression of HMGA2 plays a major role in leiomyomas, we analyzed the molecular relationship of HMGA2 and Let-7s, both in vitro and in vivo. We first characterized that exogenous Let 7 microRNAs could directly repress the dominant transcript of HMGA2, HMGA2a. This repression was also identified for two cryptic HMGA2 transcripts in primary leiomyoma cultures. Second, we found that the endogenous Let-7s were biologically active and played a major role in the regulation of HMGA2. Then, we illustrated that Let-7 repression of HMGA2 inhibited cellular proliferation. Finally, we examined the expression levels of Let-7c and HMGA2 in a large cohort of leiomyomas (n = 120), and we found high levels of Let-7 and low levels of HMGA2 in small leiomyomas, and low levels of Let-7 and high levels of HMGA2 in large leiomyomas. Our findings suggest that the Let-7-mediated repression of HMGA2 mechanism can be an important molecular event in leiomyoma growth. PMID- 18403646 TI - Separation of splenic red and white pulp occurs before birth in a LTalphabeta independent manner. AB - For the formation of lymph nodes and Peyer's patches, lymphoid tissue inducer (LTi) cells are crucial in triggering stromal cells to recruit and retain hematopoietic cells. Although LTi cells have been observed in fetal spleen, not much is known about fetal spleen development and the role of LTi cells in this process. Here, we show that LTi cells collect in a periarteriolar manner in fetal spleen at the periphery of the white pulp anlagen. Expression of the homeostatic chemokines can be detected in stromal and endothelial cells, suggesting that LTi cells are attracted by these chemokines. As lymphotoxin (LT)alpha1beta2 can be detected on B cells but not LTi cells in neonatal spleen, starting at 4 days after birth, the earliest formation of the white pulp in fetal spleen occurs in a LTalpha1beta2-independent manner. The postnatal development of the splenic white pulp, involving the influx of T cells, depends on LTalpha1beta2 expressed by B cells. PMID- 18403648 TI - Rear-view mirrors and crystal balls: a brief reflection on drug discovery. PMID- 18403647 TI - Lysophospholipid metabolism facilitates Toll-like receptor 4 membrane translocation to regulate the inflammatory response. AB - Sepsis, an overwhelming inflammatory response to infection, is a major cause of morbidity and mortality worldwide and has no specific therapy. Phospholipid metabolites, such as lysophospholipids, have been shown to regulate inflammatory responses in sepsis, although their mechanism of action is not well understood. The phospholipid-metabolizing enzymes, lysophospholipid acyltransferases, control membrane phospholipid composition, function, and the inflammatory responses of innate immune cells. Here, we show that lysophosphatidylcholine acyltransferase (LPCAT) regulates inflammatory responses to LPS and other microbial stimuli. Specific inhibition of LPCAT down-regulated inflammatory cytokine production in monocytes and epithelial cells by preventing translocation of TLR4 into membrane lipid raft domains. Our observations demonstrate a new regulatory mechanism that facilitates the innate immune responses to microbial molecular patterns and provide a basis for the anti-inflammatory activity observed in many phospholipid metabolites. This provides the possibility of the development of new classes of anti-inflammatory and antisepsis agents. PMID- 18403649 TI - Obesity, body image and diet drugs: 100 years of change. PMID- 18403651 TI - An old dog learns a new trick: regulation of peripheral glucose homeostasis by the serotonin (5-HT)2C receptor. PMID- 18403652 TI - Taste cells of the gut and gastrointestinal chemosensation. PMID- 18403653 TI - The obesity epidemic: pharmacological challenges. AB - Obesity, defined by a body mass index greater than 30kg/m(2), claims an increasing number of lives every year, underscoring a dire need for effective therapeutic interventions. The origins of the obesity epidemic are complex, but commonly cited factors include the large quantities of calorie-rich food that are readily accessible in modern society; eating habits adapted to fast-paced lifestyles; low levels of physical activity; and genetic programs that have evolved, especially in populations prone to famine, to favor the storage of excess calories (i.e., the thrifty-gene theory). It is estimated that more than thirty percent of adults, and about fifteen percent of juveniles, are obese. These high rates have led to dramatic increases in diseases such as type 2 diabetes, cardiovascular and respiratory diseases, depression, and some forms of cancer. PMID- 18403654 TI - Schizophrenia: moving beyond monoamine antagonists. AB - Schizophrenia is a disabling psychiatric disorder characterized by positive, negative, and cognitive symptoms. The first pharmacological treatments for schizophrenia were discovered by serendipitous, albeit carefully documented, clinical observations. The discovery of chlorpromazine and other dopamine D2 receptor antagonists as antipsychotic agents set the early course of drug discovery in the context of schizophrenia and other psychiatric disorders, and various monoamine receptors became the prime focus of neuropharmacological studies. Success in treating the positive symptoms nevertheless remained limited by the general lack of efficacy in addressing negative symptoms and cognitive impairment. In recent years, several new experimental approaches have emerged for the identification and treatment of different symptom clusters that do not rely on blockade of monoamine receptors. Muscarinic, nicotinic, and glutamatergic signaling mechanisms have become essential to neuropharmacological and behavioral models of discrete aspects of schizophrenia. And as a consequence of these insights, novel drug entities have become available to study and potentially treat the disabling cognitive and negative symptoms of psychiatric disease. Current attempts to target a new range of receptors entail unprecedented fine tuning in the pharmacological manipulation of specific receptor subtypes. PMID- 18403655 TI - Yellow nail syndrome: analysis of 41 consecutive patients. AB - BACKGROUND: Yellow nail syndrome (YNS) is a rare condition defined by the presence of yellow nails associated with lymphedema and/or chronic respiratory manifestations. Several aspects of this disorder remain poorly defined. METHODS: We sought to clarify the clinical features and course associated with YNS by analyzing 41 consecutive cases evaluated at a tertiary referral medical center. RESULTS: There were 20 men and 21 women; median age at diagnosis was 61 years (range, 18 to 82 years). None had a family history of YNS. All but one patient had chronic respiratory manifestations that included pleural effusions (46%), bronchiectasis (44%), chronic sinusitis (41%), and recurrent pneumonias (22%); 26 patients (63%) had lymphedema. Treatment included rotating antibiotic therapy for bronchiectasis, thoracenteses, oral vitamin E, and corticosteroid therapy. Eight patients underwent surgical management of recurrent pleural effusions including pleurodesis and decortication; two additional patients underwent pleurodesis via tube thoracostomy. The yellow nails improved or resolved in 14 of 25 patients (56%) for whom relevant data were available. Median survival of this cohort using the Kaplan-Meier method was 132 months, significantly lower than (p = 0.01) the control population. Among those still alive (20 patients), the disease appeared stable. CONCLUSIONS: In most cases, YNS is an acquired disorder and associated respiratory manifestations are generally manageable with a regimen of medical and surgical treatments. Yellow nails improve in about one half of patients, often without specific therapy. PMID- 18403656 TI - Clinically significant interstitial lung disease in limited scleroderma: histopathology, clinical features, and survival. AB - PURPOSES: To evaluate the pathologic patterns, clinical features, and survival among subjects with scleroderma (ie, systemic sclerosis [SSc]) and clinically significant interstitial lung disease (ILD) evaluated at an ILD center. METHODS: Retrospective cohort study of all SSc patients who had been referred for further evaluation of ILD and had undergone surgical lung biopsy. Clinical data were abstracted by review of the medical record, and lung biopsy specimens were reviewed and classified according to current pathologic criteria. RESULTS: All patients presented with significant respiratory symptoms. Twenty-two of 27 subjects had surgical lung biopsy-proven ILD, and 5 subjects had miscellaneous non-ILD patterns. Of those subjects with ILD, 64% (14 of 22 subjects) had a nonspecific interstitial pneumonia (NSIP) pathologic pattern (fibrotic NSIP, 13 subjects; cellular NSIP, 1 subject), and 36% (8 of 22 subjects) had the usual interstitial pneumonia (UIP) pattern. Subjects with NSIP were younger (median age, 42 vs 58 years, respectively; p = 0.003), but no differences were noted in pulmonary physiology (FVC: NSIP group, 52% predicted; UIP group, 65% predicted; p = 0.22; diffusing capacity of the lung for carbon monoxide: NSIP group, 40% predicted; UIP group, 42% predicted; p = 1.0). All patients had limited skin involvement. The Scl-70 antibody was absent among those assessed (NSIP group, 0 of 10 subjects; UIP group, 0 of 7 subjects). All patients were treated with cytotoxic therapy. The median survival time for those with NSIP was 15.3 years (5,596 days) compared with 3 years (1,084 days) for those with UIP (p = 0.07 [log rank test]). CONCLUSIONS: In SSc patients with limited cutaneous disease and clinically significant ILD, fibrotic NSIP and UIP are the predominant pathologic patterns. Those with the UIP pattern of disease had a trend toward shorter survival time. PMID- 18403657 TI - Variation in ICU risk-adjusted mortality: impact of methods of assessment and potential confounders. AB - BACKGROUND: Federal and state agencies are considering ICU performance assessment and public reporting; however, an accurate method for measuring performance must be selected. In this study, we determine whether a substantial variation in ICU mortality performance still exists in modern ICUs, and compare the predictive accuracy, reliability, and data burden of existing ICU risk-adjustment models. METHODS: A retrospective chart review of 11,300 ICU patients from 35 California hospitals from 2001 to 2004 was performed. We calculated standardized mortality ratios (SMRs) for each hospital using the mortality probability model III (MPM(0) III), the simplified acute physiology score (SAPS) II, and the acute physiology and chronic health evaluation (APACHE) IV risk-adjustment models. We compared discrimination, calibration, data reliability, and abstraction time for the models. RESULTS: Regardless of the model used, there was a large variation in SMRs among the ICUs studied. The discrimination and calibration were adequate for all risk-adjustment models. APACHE IV had the best discrimination (area under the receiver operating characteristic curve [AUC], 0.892) compared to MPM(0) III (AUC, 0.809), and SAPS II (AUC, 0.873; p < 0.001). The models differed substantially in data abstraction times, as follows: MPM(0)III, 11.1 min (95% confidence interval [CI], 8.7 to 13.4); SAPS II, 19.6 min (95% CI, 17.0 to 22.2); and APACHE IV, 37.3 min (95% CI, 28.0 to 46.6). CONCLUSIONS: We found substantial variation in the ICU risk-adjusted mortality rates that persisted regardless of the risk-adjustment model. With unlimited resources, the APACHE IV model offers the best predictive accuracy. If constrained by cost and manual data collection, the MPM(0) III model offers a viable alternative without a substantial loss in accuracy. PMID- 18403658 TI - Effects of telithromycin in in vitro and in vivo models of lipopolysaccharide induced airway inflammation. AB - BACKGROUND: The ketolide antibiotic telithromycin (TEL) exerts immunomodulatory and antiinflammatory effects in vitro and in a mouse model of septic shock. We studied the antiinflammatory activity of TEL in in vitro and in vivo models of airway inflammation induced by lipopolysaccharide (LPS). METHODS: We measured the effects of TEL on the response of RAW 264.7 macrophages to LPS and of murine lung epithelial (MLE)-12 cells to supernatants of LPS-stimulated RAW 264.7 macrophages. Macrophage inflammatory protein (MIP)-2 and tumor necrosis factor (TNF)-alpha production, nuclear factor (NF)-kappaB activation, and apoptosis were determined. Acute airway inflammation was induced in untreated and TEL-treated BALB/c mice by nebulization with LPS. Total number of leukocytes, macrophages, and neutrophils, the protein concentration, and nitrite and cytokine levels were determined in the BAL fluid. RESULTS: TEL inhibited in a dose-dependent manner the production of MIP-2 and TNF-alpha by LPS-stimulated RAW 264.7 macrophages, and the production of MIP-2 by MLE-12 epithelial cells to supernatants of LPS stimulated RAW 264.7 macrophages. NF-kappaB activation was inhibited and apoptosis was increased in both cell lines by TEL. The LPS-induced influx of neutrophils in BAL fluid was decreased by TEL pretreatment. TEL also reduced protein, nitrite, MIP-2, and TNF-alpha levels in the BAL fluid of LPS-nebulized animals. CONCLUSIONS: We have provided evidence that TEL exerts potent antiinflammatory effects in LPS-induced airways injury. We propose that TEL acts in the early phase of inflammation by reducing the release of inflammatory mediators through NF-kappaB inhibition, and in the later phase through enhancement of inflammatory cell apoptosis. PMID- 18403659 TI - Tracheostomy tube malposition in patients admitted to a respiratory acute care unit following prolonged ventilation. AB - BACKGROUND: Tracheostomy tube malposition is a barrier to weaning from mechanical ventilation. We determined the incidence of tracheostomy tube malposition, identified the associated risk factors, and examined the effect of malposition on clinical outcomes. METHODS: We performed a retrospective study on 403 consecutive patients with a tracheostomy who had been admitted to an acute care unit specializing in weaning from mechanical ventilation between July 1, 2002, and December 31, 2005. Bronchoscopy reports were reviewed for evidence of tracheostomy tube malposition (ie, > 50% occlusion of lumen by tissue). The main outcome parameters were the incidence of tracheostomy tube malposition; demographic, clinical, and tracheostomy-related factors associated with malposition; clinical response to correct the malposition; the duration of mechanical ventilation; the length of hospital stay; and mortality. RESULTS: Malpositioned tracheostomy tubes were identified in 40 of 403 patients (10%). The subspecialty of the surgical service physicians who performed the tracheostomy was most strongly associated with malposition. Thoracic and general surgeons were equally likely to have their patients associated with a malpositioned tracheostomy tube, while other subspecialty surgeons were more likely (odds ratio, 6.42; 95% confidence interval, 1.82 to 22.68; p = 0.004). Malpositioned tracheostomy tubes were changed in 80% of cases. Malposition was associated with prolonged mechanical ventilation posttracheostomy (median duration, 25 vs 15 d; p = 0.009), but not with increased hospital length of stay or mortality. CONCLUSION: Tracheostomy tube malposition appears to be a common and important complication in patients who are being weaned from mechanical ventilation. Surgical expertise may be an important factor that impacts this complication. PMID- 18403660 TI - High-resolution CT findings of parenchymal fibrosis correlate with prognosis in hypersensitivity pneumonitis. AB - BACKGROUND: Histopathologic evidence of fibrosis on surgical lung biopsy has been associated with reduced survival in patients with hypersensitivity pneumonitis (HP). Changes of pulmonary fibrosis detected on CT may also correlate with prognosis in patients with HP. METHODS: We identified 69 consecutive patients with HP diagnosed between January 1997 and December 2002 at Mayo Clinic, Rochester, MN. Patients were stratified into fibrotic and nonfibrotic groups based on the CT findings. Fibrosis was defined by the presence of irregular linear opacities, traction bronchiectasis, or honeycombing. MEASUREMENTS AND MAIN RESULTS: Of 69 patients, 26 were classified as fibrotic and 43 as nonfibrotic. Patients in the fibrotic group were older, had longer symptom duration, were more likely to have crackles on auscultation, more likely to be exposed to avian antigen, and had greater restrictive lung impairment (p<0.05 for all comparisons). There were 11 deaths in the fibrotic group and 1 death in the nonfibrotic group (p<0.0001). In the regression analysis, CT evidence of fibrosis, more severe pulmonary function abnormalities, and the presence of crackles on auscultation were predictive of reduced survival (p<0.05 for all). The presence as well as the extent of fibrosis on CT was associated with increased mortality. The age-adjusted hazard ratio for mortality in patients with fibrosis was 4.6 (95% confidence interval, 2.0 to 20.1; p<0.0001). CONCLUSION: CT findings of parenchymal fibrosis are associated with reduced survival in patients with HP and may serve as a useful prognostic indicator. PMID- 18403661 TI - COPD stage and risk of hospitalization for infectious disease. AB - BACKGROUND: Respiratory tract infections are a frequent complication of COPD, but little is known about the incidence, association, and risk of infectious diseases related to impaired lung function. METHODS: Participants in the Copenhagen City Heart Study had lung function measured at baseline, and were followed for up to 25 years. All hospitalizations due to any infection were identified through registry linkage. Impaired lung function was defined according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) staging system. MAIN RESULTS: A total of 3,333 infectious disease hospitalizations (IDHs) occurred during 230,653 person-years of follow-up (PY), corresponding to an overall incidence of 145 IDHs (95% confidence interval [CI], 139 to 149) per 10,000 PY. The incidence increased with GOLD stage, from 131 (95% CI, 126 to 136) for normal COPD, to 170 (95% CI, 146 to 193) for mild COPD, to 230 (95% CI, 207 to 253) for moderate COPD, and 394 (95% CI, 330 to 459) for severe/very severe COPD. The trend in risk persisted after the control of background characteristics using Cox proportional hazards analysis (adjusted relative risks: 1.06 [95% CI, 0.92 to 1.23], 1.39 [95% CI, 1.24 to 1.56], and 2.21 [95% CI, 1.84 to 2.64], respectively; p=0.001). In subgroup analysis, the increased risk was associated with lower and upper respiratory tract infections, pyothorax, and tuberculosis, but not with influenza, sepsis, skin infections, urinary tract infections, diarrheal disease, or other infectious diseases. CONCLUSIONS: The presence of obstructive lung disease is a significant predictor of IDH caused by respiratory tract infections, but not of hospitalizations due to infections outside the respiratory system. PMID- 18403662 TI - Short-term effects of cardiac resynchronization therapy on sleep-disordered breathing in patients with systolic heart failure. AB - OBJECTIVES: We evaluated the short-term effect of cardiac resynchronization therapy (CRT) on sleep apnea in patients with systolic heart failure. BACKGROUND: Sleep-disordered breathing is common in patients with left ventricular systolic dysfunction. METHODS: Twelve patients (mean [+/-SE] age, 59.6+/-7.8 years; mean left ventricular ejection fraction, 28.0+/-2.8%) with an implanted atrial synchronized biventricular pacemaker for the treatment of left ventricular systolic dysfunction were selected and studied. Each subject underwent polysomnography on 3 consecutive nights with CRT on the first night, CRT off the second night, and CRT on the third night. Echocardiography was performed prior to each polysomnogram. RESULTS: The central sleep event index (ie, the number of central sleep apneas [CSAs] and hypopneas per hour of sleep) score was lower with CRT compared to that without CRT (mean central sleep event index score with CRT on, 6.9+/-1.7 events per hour of sleep; mean central sleep event index score with CRT off, 14.3+/-2.9 events per hour of sleep; mean central sleep event index score with CRT on, 8.1+/-1.5 events per hour of sleep; p<0.001). Similarly, the cumulative duration of central sleep events (the number of minutes per hour of sleep during CRT) was one half that observed without CRT (CRT on, 2.8+/-0.7 min per hour of sleep; CRT OFF 6.2+/-1.2 min per hour of sleep; CRT ON 3.1+/-0.7 min per hour of sleep; p<0.001). There was a significant correlation between mitral regurgitant volume and central sleep event index on all three nights (r>or=0.77; p<0.01). CONCLUSIONS: CRT reduces CSA severity in the short term. This reduction correlated significantly with the CRT-mediated reduction of mitral regurgitation. PMID- 18403663 TI - Structural and functional lung disease in primary ciliary dyskinesia. AB - BACKGROUND: High-resolution CT (HRCT) scan data on primary ciliary dyskinesia (PCD) related lung disease are scarce. STUDY OBJECTIVES: We evaluated the lung disease in children and adults with PCD by a modified Brody composite HRCT scan score to assess the prevalence of the structural abnormalities; to evaluate the correlation among HRCT scan scores, spirometry findings, and clinical data; and to compare the PCD scores with those of age-matched and sex-matched cystic fibrosis (CF) patients. PATIENTS AND METHODS: Twenty PCD patients (age range, 4.6 to 27.5 years) underwent HRCT scanning, spirometry, and deep throat or sputum culture. A modified Brody score was used to assess bronchiectasis, mucous plugging, peribronchial thickening, parenchyma abnormalities, and mosaic perfusion. RESULTS: The total HRCT scan score was 6% of the maximal score (range, 0.5 to 25.5). Subscores were as follows: bronchiectasis, 5.6%; mucous plugging, 5.6%; peribronchial thickening, 8.3%; parenchyma, 3%; and mosaic perfusion, 0%. The prevalence of lung changes were as follows: bronchiectasis, 80%; peribronchial thickening, 80%; mucous plugging, 75%; parenchyma, 65%; and mosaic perfusion, 45%. Sixteen of 19 PCD patients had positive culture findings, and the most common pathogen found was Haemophilus influenzae (84%). The total HRCT scan score was significantly related to age (p = 0.006), FEV(1) (p = 0.02), and FVC (p = 0.02). The bronchiectasis subscore was significantly related to FEV(1) (p = 0.04) and FVC (p = 0.03). In CF patients, the total HRCT scan score was significantly higher than that in PCD patients (p = 0.02). CONCLUSIONS: PCD patients show significantly lower pulmonary HRCT scan scores than CF patients. The PCD total and bronchiectasis scores correlate with spirometry findings. The PCD HRCT scan score might be used for longitudinal assessment and/or represent an outcome surrogate in future studies. PMID- 18403664 TI - Relevance of lung ultrasound in the diagnosis of acute respiratory failure: the BLUE protocol. AB - BACKGROUND: This study assesses the potential of lung ultrasonography to diagnose acute respiratory failure. METHODS: This observational study was conducted in university-affiliated teaching-hospital ICUs. We performed ultrasonography on consecutive patients admitted to the ICU with acute respiratory failure, comparing lung ultrasonography results on initial presentation with the final diagnosis by the ICU team. Uncertain diagnoses and rare causes (frequency<2%) were excluded. We included 260 dyspneic patients with a definite diagnosis. Three items were assessed: artifacts (horizontal A lines or vertical B lines indicating interstitial syndrome), lung sliding, and alveolar consolidation and/or pleural effusion. Combined with venous analysis, these items were grouped to assess ultrasound profiles. RESULTS: Predominant A lines plus lung sliding indicated asthma (n=34) or COPD (n=49) with 89% sensitivity and 97% specificity. Multiple anterior diffuse B lines with lung sliding indicated pulmonary edema (n=64) with 97% sensitivity and 95% specificity. A normal anterior profile plus deep venous thrombosis indicated pulmonary embolism (n=21) with 81% sensitivity and 99% specificity. Anterior absent lung sliding plus A lines plus lung point indicated pneumothorax (n=9) with 81% sensitivity and 100% specificity. Anterior alveolar consolidations, anterior diffuse B lines with abolished lung sliding, anterior asymmetric interstitial patterns, posterior consolidations or effusions without anterior diffuse B lines indicated pneumonia (n=83) with 89% sensitivity and 94% specificity. The use of these profiles would have provided correct diagnoses in 90.5% of cases. CONCLUSIONS: Lung ultrasound can help the clinician make a rapid diagnosis in patients with acute respiratory failure, thus meeting the priority objective of saving time. PMID- 18403665 TI - Pleural effusion after ventricular assist device placement: prevalence and pleural fluid characteristics. AB - BACKGROUND: The occurrence of pleural effusion after ventricular assist device (VAD) implantation has been described; however, little has been elucidated about the nature, prevalence, or characteristics of the effusions. Our study details the prevalence of pleural effusion and pleural fluid characteristics in VAD patients at our institution. METHODS: We conducted a review of 22 consecutive patients undergoing VAD placement from August 2004 to January 2006. The clinical course of pleural effusions and their biochemical characteristics were studied by reviewing the patient charts and radiographs. RESULTS: Six of the 22 patients (18%) had pleural effusion before VAD placement. All 22 patients had effusions after VAD placement, with the majority being left sided (23%) or bilateral with left-sided predominance (41%). Four patients had large effusion, nine patients had moderate-sized effusions, and nine patients had small effusions. Nine patients (41%) required thoracentesis to relieve dyspnea. All were noted to have blood-tinged pleural fluid, and removal resulted in relief of dyspnea and improvement of clinical status. Seven patients had their pleural fluid examined in detail, and all met criteria for an exudate. No complications were experienced from thoracentesis. CONCLUSIONS: Although pleural effusion is commonly seen in patients after VAD placement, this is the first study to examine the effusions in detail. In our series, pleural effusions developed in all patients, and most were either on the left side or bilateral. Those sampled were exudative in nature, blood tinged, and lymphocyte predominant. Drainage resulted in improvement in symptoms and was accomplished without complications. PMID- 18403666 TI - Leptin, a novel predictor of lung function in heart failure. AB - BACKGROUND: Leptin is a protein hormone produced by adipose tissue. Leptin has proinflammatory properties and is usually elevated in patients with chronic heart failure. We assessed if serum leptin relates to the loss in lung function in noncachectic patients with chronic heart failure. MATERIALS AND METHODS: One hundred thirty-five consecutively eligible non-Hispanic white subjects (age, 24 to 79 years; 85 men and 50 women) with a diagnosis of stable systolic heart failure were recruited prospectively, along with 106 matched control subjects. FVC, FEV(1), and single-breath diffusing capacity of the lung for carbon monoxide (DLco) were measured by spirometry. Plasma leptin was measured by radioimmunoassay. Multiple linear regression was applied. RESULTS: The relationships of FEV(1), FVC, and DLco with leptin differed significantly between heart failure and control subjects after controlling for age, sex, percentage of body fat, and ejection fraction. In heart failure, leptin was as an independent predictor of FVC values (additional R(2) = 0.05, p < 0.0001), FEV(1) values (additional R(2) = 0.05, p < 0.0001), and DLco values (additional R(2) = 0.14, p < 0.0001). In a final multiple regression model predicting lung function in heart failure, the independent effect of leptin was significant after further adjustments. CONCLUSIONS: The predictive information provided by leptin is additive to that provided by measures of body fat in heart failure patients, especially for DLco. Leptin may play a role in the impairment of lung function in subjects with heart failure. PMID- 18403667 TI - Are patients with COPD more active after pulmonary rehabilitation? AB - BACKGROUND: Despite a variety of benefits brought by pulmonary rehabilitation to patients with COPD, it is unclear whether these patients are more active during daily life after the program. METHODS: Physical activities in daily life (activity monitoring), pulmonary function (spirometry), exercise capacity (incremental cycle-ergometer testing and 6-min walk distance testing), muscle force (quadriceps and handgrip force, and inspiratory and expiratory maximal pressures), quality of life (chronic respiratory disease questionnaire), and functional status (pulmonary functional status and dyspnea questionnaire-modified version) were assessed at baseline, after 3 months of a multidisciplinary rehabilitation program, and at the end of a 6-month multidisciplinary rehabilitation program in 29 patients (mean [+/- SD] age, 67 +/- 8 years; FEV(1), 46 +/- 16% predicted). RESULTS: Exercise capacity, muscle force, quality of life, and functional status improved significantly after 3 months of pulmonary rehabilitation (all p < 0.05), with further improvements in muscle force, functional status, and quality of life at 6 months. Movement intensity during walking improved significantly after 3 months (p = 0.046) with further improvements after 6 months (p = 0.0002). Walking time in daily life did not improve significantly at 3 months (mean improvement, 7 +/- 35%; p = 0.21), but only after 6 months (mean improvement, 20 +/- 36%; p = 0.008). No significant changes occurred in other activities or in the pattern of the time spent walking in daily life. Changes in dyspnea after the program were significantly related to changes in walking time in daily life (r = 0.43; p = 0.02). CONCLUSION: If one aims at changing physical activity habits in the daily life of COPD patients, the contribution of long-lasting programs might be important. PMID- 18403668 TI - Rapid effect of inhaled ciclesonide in asthma: a randomized, placebo-controlled study. AB - BACKGROUND: Ciclesonide is a novel inhaled corticosteroid for the treatment of asthma, and it is important to measure the onset of effect of this therapy on airway hyperresponsiveness (AHR), exhaled nitric oxide (NO), and levels of eosinophils in induced sputum. METHODS: In a randomized, double-blind, crossover study, 21 patients with mild asthma inhaled ciclesonide 320 microg (ex-actuator) qd, ciclesonide 640 microg (ex-actuator) bid, and placebo for 7 days. Exhaled NO and AHR to adenosine monophosphate (AMP), measured as the provocative concentration of AMP producing a 20% reduction in FEV1 (PC20FEV1), were assessed after inhalation on days 1, 3 and 7. Eosinophil levels in induced sputum were also measured. RESULTS: Ciclesonide 320 microg qd and 640 microg bid produced significantly greater improvements in PC20FEV1 compared with placebo on day 1 (within 2.5 h), and on days 3 and 7 (all p < 0.0001). On day 3, both ciclesonide doses significantly reduced exhaled NO levels by - 17.7 parts per billion (p < 0.0001) and - 15.4 parts per billion (p < 0.003) vs placebo, respectively. Significant reductions were maintained during the study with both ciclesonide doses (p < 0.01). A nonsignificant trend towards a decrease in eosinophil cell numbers was observed after 7 days of ciclesonide treatment, especially in patients receiving the higher dose. CONCLUSIONS: A single dose of ciclesonide decreased AHR to AMP and exhaled NO within 3 h, while FEV, improved at 3 days and 7 days. PMID- 18403669 TI - Prognostic role of clinical and laboratory criteria to identify early ventilator associated pneumonia in brain injury. AB - BACKGROUND: We investigated the role of the clinical pulmonary infection score (CPIS), serum levels of procalcitonin (PCT), C-reactive protein (CRP), and serum amyloid A (SAA) in the detection of patients with early ventilator-associated pneumonia (VAP). METHODS: Observational study in a university hospital. In 58 patients with severe brain injury receiving mechanical ventilation, CPIS, PCT, CRP and SAA were evaluated at ICU entry and at days 3 to 4 of hospital stay for VAP diagnosis (confirmed by endotracheal aspirate or BAL cultures). RESULTS: We found the following: (1) PCT at entry was increased in patients who later had early VAP develop (25 patients) compared to no VAP (median, 1.4 ng/mL; 25-75 percentiles, 0.14-0.78; vs median, 0.2 ng/mL; 25-75 percentiles, 0.76-2.4, p<0.001; sensitivity, 76%; and specificity, 75%); (2) CPIS increased at the day of VAP diagnosis, compared to entry (median, 6.6+/-1.1 vs 1.5+/-1.1, p<0.001; sensitivity, 97%; specificity, 100%), while other serum inflammatory markers did not change; and (3) deterioration in oxygenation and changes in tracheal secretions were the main determinants of CPIS changes. CONCLUSIONS: PCT may be a useful marker to predict which patients subsequently have early VAP. The CPIS could help as an early way to detect the patients with early VAP and who need further diagnostic testing. PMID- 18403670 TI - Exercise-induced pulmonary arterial hypertension in patients with systemic sclerosis. AB - INTRODUCTION: Pulmonary arterial hypertension (PAH) is the most common cause of scleroderma-related deaths. New medications for PAH patients make it necessary to identify patients with high risk factors for PAH. This study looks at the use of an exercise echocardiogram in identifying patients who may have PAH and may be candidates for early therapeutic intervention. METHODS: This study included 54 scleroderma patients with symptoms suggesting they were at risk for pulmonary hypertension, including dyspnea on exertion, diffusing capacity of the lung for carbon monoxide (Dlco)<60% of predicted, FVC<70% of predicted, percentage of predicted FVC/percentage of predicted Dlco (FVC%/Dlco%) ratio>1.6, or resting right ventricular systolic pressure (RVSP)>35 mm Hg. The exercise echocardiogram protocol involved the standard Bruce stress echocardiogram protocol with remeasurement of the RVSP within 1 min of stopping exercise. A positive exercise test result was defined as an increase of at least 20 mm Hg in the RVSP with exercise. Right-heart catheterization with exercise was performed in those with a positive exercise test result. RESULTS: Resting mean RVSP was 34.5 mm Hg, which increased to 51.4 mm Hg with exercise; 44% had at a positive exercise test result, which correlated with a low Dlco, high FVC%/Dlco% ratio (p<0.001), a positive anti-centromere antibody, and RVSP>35 mm Hg (p<0.05). PAH was confirmed by right-heart catheterization in 81% of patients: 19% at rest and 62% of patients with exercise. CONCLUSIONS: Exercise-induced pulmonary hypertension is a common finding in patients at high risk for PAH. This may be a sensitive way to identify patients with early PAH. Long-term follow-up and early treatment should be studied in these patients. PMID- 18403671 TI - FEV1 response to bronchodilation in an adult urban population. AB - BACKGROUND: Most studies evaluating bronchodilation in flow-volume spirometry have been conducted in patients with obstructive airways diseases, but less is known about bronchodilation responses in the general population or in healthy subjects. METHODS: We evaluated an urban population sample of 628 adults (260 men, 368 women) aged 25 to 74 years with flow-volume spirometry using inhalation of 0.4 mg of a salbutamol aerosol with a spacer device for bronchodilation. On the basis of a structured interview, a subgroup of 219 healthy, asymptomatic nonsmokers was selected. RESULTS: In the population sample, the average increase in FEV(1) from baseline after salbutamol inhalation was 77.2 mL (SD, 109.7 mL) or 2.5% (SD, 3.9%). In healthy asymptomatic nonsmokers, the mean change in FEV(1) was 62.0 mL (SD, 89.7 mL) or 1.8% (SD, 2.6%). In the whole population, the 95th percentile limit of the increase in FEV(1) was 8.5%, while it was 5.9% among healthy asymptomatic nonsmokers. The absolute change in FEV(1) correlated significantly with baseline FVC (p < 0.01). The FEV(1)/FVC ratio at baseline was the strongest influencing factor for the bronchodilation response. CONCLUSIONS: The results indicate that a significant increase in FEV(1) from baseline in a bronchodilation test is around 9% in an urban population. The level of the significant absolute increase in FEV(1) seems to depend on FVC. Low baseline FEV(1)/FVC ratio, reflecting airflow limitation, is the strongest determinant for FEV(1) response to bronchodilation. PMID- 18403672 TI - Comparison of a combination of tiotropium plus formoterol to salmeterol plus fluticasone in moderate COPD. AB - BACKGROUND: A 6-week, multicenter, randomized, double-blind, parallel-group study was conducted in patients with COPD to compare lung function improvements of tiotropium, 18 microg qd, plus formoterol, 12 microg bid, to salmeterol, 50 microg bid, plus fluticasone, 500 microg bid. METHODS: Following a screening visit, subjects entered a run-in period in which they received regular ipratropium. At randomization, patients were assigned to either tiotropium plus formoterol or salmeterol plus fluticasone. After 6 weeks of treatment, a 12-h lung function profile was obtained. The coprimary end points were FEV(1) area under the curve for the time period 0 to 12 h (AUC(0-12)) and peak FEV(1). RESULTS: A total of 729 patients were screened, and 605 patients were randomized and treated. A total of 592 patients (baseline FEV(1), 1.32 +/- 0.43 L/min [+/ SD]) were included in the analysis. After 6 weeks, the 12-h lung function profiles in the group receiving tiotropium plus formoterol were superior to those in the group receiving salmeterol plus fluticasone (mean difference in FEV(1) AUC(0-12), 78 mL [p = 0.0006]; mean difference in FVC AUC(0-12), 173 mL, p < 0.0001). Also, peak responses were in favor of tiotropium plus formoterol (difference in peak FEV(1), 103 mL [p < 0.0001]; difference in peak FVC, 214 mL [p < 0.0001]), as were FEV(1) and FVC at each individual time point after dose (p < 0.05). Predose FVC was significantly higher with the bronchodilator combination, while predose FEV(1) and rescue medication use did not differ significantly between groups. Both treatments were well tolerated. CONCLUSIONS: Tiotropium plus formoterol was superior in lung function over the day compared to salmeterol plus fluticasone in patients with moderate COPD. Long-term studies in patients with severe COPD are warranted to assess the relative efficacy of different treatment combinations. TRIAL REGISTRATION: Clinicaltrials.gov Identifier: NCT00239421. PMID- 18403673 TI - Treprostinil-based therapy in the treatment of moderate-to-severe pulmonary arterial hypertension: long-term efficacy and combination with bosentan. AB - BACKGROUND: Treprostinil, a long-acting prostacyclin analog, diminished the symptoms of pulmonary arterial hypertension (PAH) in controlled 12-week clinical efficacy studies. This retrospective, single-center, open-label study was designed to assess the efficacy of long-term, subcutaneously administered, treprostinil-based therapy alone or in combination with bosentan for the treatment of moderate-to-severe PAH. METHODS: Thirty-eight patients with pulmonary hypertension treated with subcutaneous treprostinil were followed up for a mean (+/-SD) duration of 984+/-468 days (range, 165 to 1,847 days). Oral bosentan was added to the treprostinil regimen if patients remained in New York Heart Association (NYHA) functional class III or II with intolerable prostacyclin side effects that limited therapy. Hemodynamic studies, Borg dyspnea score evaluations, 6-min walk (6MW) tests, and NYHA functional class determinations were performed at approximately 6-month intervals. RESULTS: Mean pulmonary artery pressure decreased from 59.7 to 50.5 mm Hg (p<0.001). Significant and sustained improvement in 6MW distance (p=0.022) and Borg dyspnea score (p=0.023) were observed. At the final observation, the mean dose of treprostinil was 37.8 ng/kg/min (range, 7.5 to 115 ng/kg/min). At baseline, 5% of patients were in NYHA functional class 2 or lower vs 58% at the last follow-up. Bosentan was added to the regimens of 19 patients. In those patients, significant additional improvement occurred in the pulmonary arterial pressure (p<0.001), 6MW distance (p=0.001), and Borg dyspnea scale (p=0.020) compared to baseline. CONCLUSIONS: Long-term treatment with subcutaneous treprostinil-based therapy improved functional parameters and hemodynamics in patients with moderate-to-severe PAH. In patients requiring combination therapy, the addition of oral bosentan to treprostinil-based therapy was safe, well-tolerated, and associated with further clinical improvements. PMID- 18403674 TI - Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. AB - The inhalation of airborne pollutants, such as asbestos or silica, is linked to inflammation of the lung, fibrosis, and lung cancer. How the presence of pathogenic dust is recognized and how chronic inflammatory diseases are triggered are poorly understood. Here, we show that asbestos and silica are sensed by the Nalp3 inflammasome, whose subsequent activation leads to interleukin-1beta secretion. Inflammasome activation is triggered by reactive oxygen species, which are generated by a NADPH oxidase upon particle phagocytosis. (NADPH is the reduced form of nicotinamide adenine dinucleotide phosphate.) In a model of asbestos inhalation, Nalp3-/- mice showed diminished recruitment of inflammatory cells to the lungs, paralleled by lower cytokine production. Our findings implicate the Nalp3 inflammasome in particulate matter-related pulmonary diseases and support its role as a major proinflammatory "danger" receptor. PMID- 18403675 TI - Subnanometer motion of cargoes driven by thermal gradients along carbon nanotubes. AB - An important issue in nanoelectromechanical systems is developing small electrically driven motors. We report on an artificial nanofabricated motor in which one short carbon nanotube moves relative to another coaxial nanotube. A cargo is attached to an ablated outer wall of a multiwalled carbon nanotube that can rotate and/or translate along the inner nanotube. The motion is actuated by imposing a thermal gradient along the nanotube, which allows for subnanometer displacements, as opposed to an electromigration or random walk effect. PMID- 18403676 TI - Temperature sensing by an olfactory neuron in a circuit controlling behavior of C. elegans. AB - Temperature is an unavoidable environmental cue that affects the metabolism and behavior of any creature on Earth, yet how animals perceive temperature is poorly understood. The nematode Caenorhabditis elegans "memorizes" temperatures, and this stored information modifies its subsequent migration along a temperature gradient. We show that the olfactory neuron designated AWC senses temperature. Calcium imaging revealed that AWC responds to temperature changes and that response thresholds differ depending on the temperature to which the animal was previously exposed. In the mutant with impaired heterotrimeric guanine nucleotide binding protein (G protein)-mediated signaling, AWC was hyperresponsive to temperature, whereas the AIY interneuron (which is postsynaptic to AWC) was hyporesponsive to temperature. Thus, temperature sensation exhibits a robust influence on a neural circuit controlling a memory-regulated behavior. PMID- 18403678 TI - Hybrid vigor in science. PMID- 18403677 TI - Endogenous siRNAs derived from transposons and mRNAs in Drosophila somatic cells. AB - Small interfering RNAs (siRNAs) direct RNA interference (RNAi) in eukaryotes. In flies, somatic cells produce siRNAs from exogenous double-stranded RNA (dsRNA) as a defense against viral infection. We identified endogenous siRNAs (endo-siRNAs), 21 nucleotides in length, that correspond to transposons and heterochromatic sequences in the somatic cells of Drosophila melanogaster. We also detected endo siRNAs complementary to messenger RNAs (mRNAs); these siRNAs disproportionately mapped to the complementary regions of overlapping mRNAs predicted to form double stranded RNA in vivo. Normal accumulation of somatic endo-siRNAs requires the siRNA-generating ribonuclease Dicer-2 and the RNAi effector protein Argonaute2 (Ago2). We propose that endo-siRNAs generated by the fly RNAi pathway silence selfish genetic elements in the soma, much as Piwi-interacting RNAs do in the germ line. PMID- 18403679 TI - Environmental regulation. New rules on saving wetlands push the limits of the science. PMID- 18403680 TI - Medicine. Drug bestows radiation resistance on mice and monkeys. PMID- 18403681 TI - Biomedical community. HHMI's Cech signs off on his biggest experiment. PMID- 18403682 TI - Planetary geology. An early big hit to Mars may have scarred the planet for life. PMID- 18403683 TI - Applied physics. At mixed odds, racetrack memory charges from gate. PMID- 18403684 TI - Parkinson's disease. Signs of disease in fetal transplants. PMID- 18403685 TI - Psychopharmacology. Tackling alcoholism with drugs. PMID- 18403686 TI - Plant genetics. The blue revolution, drop by drop, gene by gene. PMID- 18403687 TI - Plant genetics. Getting to the root of drought responses. PMID- 18403688 TI - Society for American Archaeology meeting. Following an asphalt trail to ancient Olmec trade routes. PMID- 18403689 TI - Society for American Archaeology meeting. Herring and nuts for the 'salmon people'. PMID- 18403690 TI - Society for American Archaeology meeting. Signs of the first whale hunters. PMID- 18403691 TI - Retraction. PMID- 18403692 TI - Vegetation's role in coastal protection. PMID- 18403693 TI - A quaternary question. PMID- 18403694 TI - Soil erosion: data say C sink. PMID- 18403695 TI - Science and government. Science and the candidates. PMID- 18403696 TI - Neuroscience. Refreshing connections. PMID- 18403697 TI - Geochemistry. Toward new uses for hematite. PMID- 18403698 TI - Neuroscience. Axons seek neighborly advice. PMID- 18403699 TI - Biochemistry. Anatomy of a fungal polyketide synthase. PMID- 18403700 TI - Biochemistry. Zooming into live cells. PMID- 18403701 TI - Retrospective. Arthur C. Clarke (1917-2008). PMID- 18403702 TI - Magnetic domain-wall racetrack memory. AB - Recent developments in the controlled movement of domain walls in magnetic nanowires by short pulses of spin-polarized current give promise of a nonvolatile memory device with the high performance and reliability of conventional solid state memory but at the low cost of conventional magnetic disk drive storage. The racetrack memory described in this review comprises an array of magnetic nanowires arranged horizontally or vertically on a silicon chip. Individual spintronic reading and writing nanodevices are used to modify or read a train of approximately 10 to 100 domain walls, which store a series of data bits in each nanowire. This racetrack memory is an example of the move toward innately three dimensional microelectronic devices. PMID- 18403703 TI - Amplification of Cretaceous warmth by biological cloud feedbacks. AB - The extreme warmth of particular intervals of geologic history cannot be simulated with climate models, which are constrained by the geologic proxy record to relatively modest increases in atmospheric carbon dioxide levels. Recent recognition that biological productivity controls the abundance of cloud condensation nuclei (CCN) in the unpolluted atmosphere provides a solution to this problem. Our climate simulations show that reduced biological productivity (low CCN abundance) provides a substantial amplification of CO2-induced warming by reducing cloud lifetimes and reflectivity. If the stress of elevated temperatures did indeed suppress marine and terrestrial ecosystems during these times, this long-standing climate enigma may be solved. PMID- 18403704 TI - Electronic origin of the inhomogeneous pairing interaction in the high-Tc superconductor Bi2Sr2CaCu2O8+delta. AB - Identifying the mechanism of superconductivity in the high-temperature cuprate superconductors is one of the major outstanding problems in physics. We report local measurements of the onset of superconducting pairing in the high-transition temperature (Tc) superconductor Bi2Sr2CaCu2O8+delta using a lattice-tracking spectroscopy technique with a scanning tunneling microscope. We can determine the temperature dependence of the pairing energy gaps, the electronic excitations in the absence of pairing, and the effect of the local coupling of electrons to bosonic excitations. Our measurements reveal that the strength of pairing is determined by the unusual electronic excitations of the normal state, suggesting that strong electron-electron interactions rather than low-energy (<0.1 volts) electron-boson interactions are responsible for superconductivity in the cuprates. PMID- 18403705 TI - Surface mobility of postsynaptic AMPARs tunes synaptic transmission. AB - AMPA glutamate receptors (AMPARs) mediate fast excitatory synaptic transmission. Upon fast consecutive synaptic stimulation, transmission can be depressed. Recuperation from fast synaptic depression has been attributed solely to recovery of transmitter release and/or AMPAR desensitization. We show that AMPAR lateral diffusion, observed in both intact hippocampi and cultured neurons, allows fast exchange of desensitized receptors with naive functional ones within or near the postsynaptic density. Recovery from depression in the tens of millisecond time range can be explained in part by this fast receptor exchange. Preventing AMPAR surface movements through cross-linking, endogenous clustering, or calcium rise all slow recovery from depression. Physiological regulation of postsynaptic receptor mobility affects the fidelity of synaptic transmission by shaping the frequency dependence of synaptic responses. PMID- 18403706 TI - Current-controlled magnetic domain-wall nanowire shift register. AB - The controlled motion of a series of domain walls along magnetic nanowires using spin-polarized current pulses is the essential ingredient of the proposed magnetic racetrack memory, a new class of potential non-volatile storage-class memories. Using permalloy nanowires, we achieved the successive creation, motion, and detection of domain walls by using sequences of properly timed, nanosecond long, spin-polarized current pulses. The cycle time for the writing and shifting of the domain walls was a few tens of nanoseconds. Our results illustrate the basic concept of a magnetic shift register that relies on the phenomenon of spin momentum transfer to move series of closely spaced domain walls. PMID- 18403707 TI - Determining chondritic impactor size from the marine osmium isotope record. AB - Decreases in the seawater 187Os/188Os ratio caused by the impact of a chondritic meteorite are indicative of projectile size, if the soluble fraction of osmium carried by the impacting body is known. Resulting diameter estimates of the Late Eocene and Cretaceous/Paleogene projectiles are within 50% of independent estimates derived from iridium data, assuming total vaporization and dissolution of osmium in seawater. The variations of 187Os/188Os and Os/Ir across the Late Eocene impact-event horizon support the main assumptions required to estimate the projectile diameter. Chondritic impacts as small as 2 kilometers in diameter should produce observable excursions in the marine osmium isotope record, suggesting that previously unrecognized impact events can be identified by this method. PMID- 18403708 TI - Aligning conservation priorities across taxa in Madagascar with high-resolution planning tools. AB - Globally, priority areas for biodiversity are relatively well known, yet few detailed plans exist to direct conservation action within them, despite urgent need. Madagascar, like other globally recognized biodiversity hot spots, has complex spatial patterns of endemism that differ among taxonomic groups, creating challenges for the selection of within-country priorities. We show, in an analysis of wide taxonomic and geographic breadth and high spatial resolution, that multitaxonomic rather than single-taxon approaches are critical for identifying areas likely to promote the persistence of most species. Our conservation prioritization, facilitated by newly available techniques, identifies optimal expansion sites for the Madagascar government's current goal of tripling the land area under protection. Our findings further suggest that high-resolution multitaxonomic approaches to prioritization may be necessary to ensure protection for biodiversity in other global hot spots. PMID- 18403710 TI - Evidence for editing of human papillomavirus DNA by APOBEC3 in benign and precancerous lesions. AB - Cytidine deaminases of the APOBEC3 family all have specificity for single stranded DNA, which may become exposed during replication or transcription of double-stranded DNA. Three human APOBEC3A (hA3A), hA3B, and hA3H genes are expressed in keratinocytes and skin, leading us to determine whether genetic editing of human papillomavirus (HPV) DNA occurred. In a study of HPV1a plantar warts and HPV16 precancerous cervical biopsies, hyperedited HPV1a and HPV16 genomes were found. Strictly analogous results were obtained from transfection experiments with HPV plasmid DNA and the three nuclear localized enzymes: hA3A, hA3C, and hA3H. Thus, stochastic or transient overexpression of APOBEC3 genes may expose the genome to a broad spectrum of mutations that could influence the development of tumors. PMID- 18403709 TI - An agonist of toll-like receptor 5 has radioprotective activity in mouse and primate models. AB - The toxicity of ionizing radiation is associated with massive apoptosis in radiosensitive organs. Here, we investigate whether a drug that activates a signaling mechanism used by tumor cells to suppress apoptosis can protect healthy cells from the harmful effects of radiation. We studied CBLB502, a polypeptide drug derived from Salmonella flagellin that binds to Toll-like receptor 5 (TLR5) and activates nuclear factor-kappaB signaling. A single injection of CBLB502 before lethal total-body irradiation protected mice from both gastrointestinal and hematopoietic acute radiation syndromes and resulted in improved survival. CBLB502 injected after irradiation also enhanced survival, but at lower radiation doses. It is noteworthy that the drug did not decrease tumor radiosensitivity in mouse models. CBLB502 also showed radioprotective activity in lethally irradiated rhesus monkeys. Thus, TLR5 agonists could potentially improve the therapeutic index of cancer radiotherapy and serve as biological protectants in radiation emergencies. PMID- 18403711 TI - Segregation of axial motor and sensory pathways via heterotypic trans-axonal signaling. AB - Execution of motor behaviors relies on circuitries effectively integrating immediate sensory feedback to efferent pathways controlling muscle activity. It remains unclear how, during neuromuscular circuit assembly, sensory and motor projections become incorporated into tightly coordinated, yet functionally separate pathways. We report that, within axial nerves, establishment of discrete afferent and efferent pathways depends on coordinate signaling between coextending sensory and motor projections. These heterotypic axon-axon interactions require motor axonal EphA3/EphA4 receptor tyrosine kinases activated by cognate sensory axonal ephrin-A ligands. Genetic elimination of trans-axonal ephrin-A --> EphA signaling in mice triggers drastic motor-sensory miswiring, culminating in functional efferents within proximal afferent pathways. Effective assembly of a key circuit underlying motor behaviors thus critically depends on trans-axonal signaling interactions resolving motor and sensory projections into discrete pathways. PMID- 18403712 TI - Convergence of Campylobacter species: implications for bacterial evolution. AB - The nature of species boundaries in bacteria remains controversial. In particular, the mechanisms of bacterial speciation and maintenance in the face of frequent genetic exchange are poorly understood. Here, we report patterns of genetic exchange that show two closely related zoonotic pathogenic species, Campylobacter jejuni and Campylobacter coli, are converging as a consequence of recent changes in gene flow. Population expansion into a novel ecological niche generated by human activity is the most probable explanation for the increase in genetic exchange between these species. Bacterial speciation can therefore occur by mechanisms analogous to those seen in metazoans, where genetic diversification and incipient speciation caused by ecological factors have been reported in several genera. PMID- 18403713 TI - Leiomodin is an actin filament nucleator in muscle cells. AB - Initiation of actin polymerization in cells requires nucleation factors. Here we describe an actin-binding protein, leiomodin, that acted as a strong filament nucleator in muscle cells. Leiomodin shared two actin-binding sites with the filament pointed end-capping protein tropomodulin: a flexible N-terminal region and a leucine-rich repeat domain. Leiomodin also contained a C-terminal extension of 150 residues. The smallest fragment with strong nucleation activity included the leucine-rich repeat and C-terminal extension. The N-terminal region enhanced the nucleation activity threefold and recruited tropomyosin, which weakly stimulated nucleation and mediated localization of leiomodin to the middle of muscle sarcomeres. Knocking down leiomodin severely compromised sarcomere assembly in cultured muscle cells, which suggests a role for leiomodin in the nucleation of tropomyosin-decorated filaments in muscles. PMID- 18403714 TI - Deconstruction of iterative multidomain polyketide synthase function. AB - PksA, which initiates biosynthesis of the environmental carcinogen aflatoxin B1, is one of the multidomain iterative polyketide synthases (IPKSs), a large, poorly understood family of biosynthetic enzymes. We found that dissection of PksA and its reconstitution from selected sets of domains allows the accumulation and characterization of advanced octaketide intermediates bound to the enzyme, permitting the reactions controlled by individual catalytic domains to be identified. A product template (PT) domain unites with the ketosynthase and thioesterase in this IPKS system to assemble precisely seven malonyl-derived building blocks to a hexanoyl starter unit and mediate a specific cyclization cascade. Because the PT domain is common among nonreducing IPKSs, these mechanistic features should prove to be general for IPKS-catalyzed production of aromatic polyketides. PMID- 18403715 TI - Voices of Mexican American caregivers for family members with cancer: on becoming stronger. AB - This grounded theory study report describes the experience of 34 Mexican American (MA) female caregivers who provided care to a family member with cancer. Caregivers identified a process of "Becoming Stronger" as a result of their caregiver role. The emerging theoretical model of female MA cancer caregiving offers evidence to change current cancer care approaches from patient-focused to family-focused care for this ethnic group. Findings suggest that changes most responsive to cultural values and likely to provide accessible and quality cancer care for MA families are those that involve active partnering with MA caregivers to prioritize the patient's cancer care. PMID- 18403716 TI - Ligand dissociation from estrogen receptor is mediated by receptor dimerization: evidence from molecular dynamics simulations. AB - Estrogen Receptor (ER) is an important target for pharmaceutical design. Like other ligand-dependent transcription factors, hormone binding regulates ER transcriptional activity. Nevertheless, the mechanisms by which ligands enter and leave ERs and other nuclear receptors remain poorly understood. Here, we report results of locally enhanced sampling molecular dynamics simulations to identify dissociation pathways of two ER ligands [the natural hormone 17beta-estradiol (E(2)) and the selective ER modulator raloxifene (RAL)] from the human ERalpha ligand-binding domain in monomeric and dimeric forms. E(2) dissociation occurs via three different pathways in ER monomers. One resembles the mousetrap mechanism (Path I), involving repositioning of helix 12 (H12), others involve the separation of H8 and H11 (Path II), and a variant of this pathway at the bottom of the ligand-binding domain (Path II'). RAL leaves the receptor through Path I and a Path I variant in which the ligand leaves the receptor through the loop region between H11 and H12 (Path I'). Remarkably, ER dimerization strongly suppresses Paths II and II' for E(2) dissociation and modifies RAL escape routes. We propose that differences in ligand release pathways detected in the simulations for ER monomers and dimers provide an explanation for previously observed effects of ER quaternary state on ligand dissociation rates and suggest that dimerization may play an important, and hitherto unexpected, role in regulation of ligand dissociation rates throughout the nuclear receptor family. PMID- 18403717 TI - Molecular determinants for the activating/blocking actions of the 2H-1,4 benzoxazine derivatives, a class of potassium channel modulators targeting the skeletal muscle KATP channels. AB - The 2H-1,4-benzoxazine derivatives are modulators of the skeletal muscle ATP sensitive-K(+) channels (K(ATP)), activating it in the presence of ATP but inhibiting it in the absence of nucleotide. To investigate the molecular determinants for the activating/blocking actions of these compounds, novel molecules with different alkyl or aryl-alkyl substitutes at position 2 of the 1,4 benzoxazine ring were prepared. The effects of the lengthening of the alkyl chain and of branched substitutes, as well as of the introduction of aliphatic/aromatic rings on the activity of the molecules, were investigated on the skeletal muscle K(ATP) channels of the rat, in excised-patch experiments, in the presence or absence of internal ATP (10(-4) M). In the presence of ATP, the 2-n-hexyl analog was the most potent activator (DE(50) = 1.08 x 10(-10) M), whereas the 2 phenylethyl was not effective. The rank order of efficacy of the openers was 2-n hexyl > or =2-cyclohexylmethyl >2-isopropyl = 2-n-butyl > or = 2-phenyl > or = 2 benzyl = 2-isobutyl analogs. In the absence of ATP, the 2-phenyl analog was the most potent inhibitor (IC(50) = 2.5 x 10(-11) M); the rank order of efficacy of the blockers was 2-phenyl > or = 2-n-hexyl > 2-n-butyl > 2-cyclohexylmethyl, whereas the 2-phenylethyl, 2-benzyl, and 2-isobutyl 1,4-benzoxazine analogs were not effective; the 2-isopropyl analog activated the K(ATP) channel even in the absence of nucleotide. Therefore, distinct molecular determinants for the activating or blocking actions for these compounds can be found. For example, the replacement of the linear with the branched alkyl substitutes at the position 2 of the 1,4-benzoxazine nucleus determines the molecular switch from blockers to openers. These compounds were 100-fold more potent and effective as openers than other KCO against the muscle K(ATP) channels. PMID- 18403718 TI - Kinetic characterization and molecular docking of a novel, potent, and selective slow-binding inhibitor of human cathepsin L. AB - A novel small molecule thiocarbazate (PubChem SID 26681509), a potent inhibitor of human cathepsin L (EC 3.4.22.15) with an IC(50) of 56 nM, was developed after a 57,821-compound screen of the National Institutes of Health Molecular Libraries Small Molecule Repository. After a 4-h preincubation with cathepsin L, this compound became even more potent, demonstrating an IC(50) of 1.0 nM. The thiocarbazate was determined to be a slow-binding and slowly reversible competitive inhibitor. Through a transient kinetic analysis for single-step reversibility, inhibition rate constants were k(on) = 24,000 M(-1)s(-1) and k(off) = 2.2 x 10(-5) s(-1) (K(i) = 0.89 nM). Molecular docking studies were undertaken using the experimentally derived X-ray crystal structure of papain/CLIK-148 (1cvz. pdb). These studies revealed critical hydrogen bonding patterns of the thiocarbazate with key active site residues in papain. The thiocarbazate displayed 7- to 151-fold greater selectivity toward cathepsin L than papain and cathepsins B, K, V, and S with no activity against cathepsin G. The inhibitor demonstrated a lack of toxicity in human aortic endothelial cells and zebrafish. In addition, the thiocarbazate inhibited in vitro propagation of malaria parasite Plasmodium falciparum with an IC(50) of 15.4 microM and inhibited Leishmania major with an IC(50) of 12.5 microM. PMID- 18403719 TI - Conformational rearrangements and signaling cascades involved in ligand-biased mitogen-activated protein kinase signaling through the beta1-adrenergic receptor. AB - In recent years, several studies have demonstrated that different ligands can have distinct efficacy profiles toward various signaling pathways through a unique receptor. For example, beta1-adrenergic compounds that are inverse agonists toward the adenylyl cyclase (AC) can display agonist activity for the mitogen-activated protein kinase (MAPK) pathway. Such a phenomenon, often termed functional selectivity, has now been clearly established for many G protein coupled receptors when considering distinct signaling output. However, the possibility that ligands could selectively engage distinct effectors to activate a single signaling output by promoting specific receptor conformations has not been extensively examined. Here, we took advantage of the fact that isoproterenol, bucindolol and propranolol (full, partial, and inverse agonists for the AC pathway, respectively) all activate MAPK through the beta1-adrenergic receptor (beta1AR) to probe such conformational-biased signaling. Although the three compounds stimulated MAPK in a src-dependent manner, isoproterenol acted through both Galpha(i)betagamma- and G protein-independent pathways, whereas bucindolol and propranolol promoted MAPK activation through the G protein independent pathway only. The existence of such distinct signaling cascades linking beta1AR to MAPK activation was correlated with ligand-specific conformational rearrangements of receptor/G protein complexes measured by bioluminescence resonance energy transfer. Taken together, our data indicate that discrete local conformational changes can selectively promote the recruitment of distinct proximal signaling partners that can engage distinct signaling outputs and/or converge on the same signaling output. PMID- 18403720 TI - Niemann-pick C1-like 1 mediates alpha-tocopherol transport. AB - Dietary lipids and fat-soluble micronutrients are solubilized in mixed micelles and absorbed in the small intestine. Based on an assumption that cholesterol and other fat-soluble molecules share a number of transport mechanisms and the fact that Niemann-Pick C1-like 1 (NPC1L1) is critical for intestinal cholesterol absorption, we hypothesized that some fat-soluble molecules may be transported by NPC1L1. To investigate this hypothesis, we compared the cellular uptake and inhibitory effects of ezetimibe, the molecular target of which is NPC1L1, between cholesterol and some fat-soluble molecules using rat NPC1L1-overexpressing Caco-2 cells. The in vitro analysis suggested that NPC1L1 mediates the uptake of alpha tocopherol (vitamin E) in an ezetimibe-sensitive manner as well as the uptake of cholesterol but does not mediate the uptake of retinol (vitamin A) or cyclosporin A. To confirm the ezetimibe-sensitive uptake of alpha-tocopherol in vivo, we performed an in vivo absorption study using rats and the results suggested a physiologically significant role of NPC1L1-mediated alpha-tocopherol absorption. Furthermore, using human NPC1L1 overexpression system, we demonstrated that both cholesterol and alpha-tocopherol uptake was also significantly increased by the overexpression of human NPC1L1 and ezetimibe inhibited their uptake. Mutual inhibition studies of cholesterol and alpha-tocopherol in human NPC1L1-mediated uptake revealed the inhibitory effect of cholesterol and the stimulatory effect of alpha-tocopherol on the NPC1L1-mediated transport of both substrates. The present data suggest, for the first time, that NPC1L1 has the ability to transport alpha-tocopherol and that ezetimibe is able to inhibit the intestinal absorption of alpha-tocopherol. PMID- 18403721 TI - Lung function from infancy to the preschool years after clinical diagnosis of cystic fibrosis. AB - RATIONALE: After recent standardization of forced expiratory maneuvers for both infants and preschool children, longitudinal measurements are now possible from birth. OBJECTIVES: The aim of this study was to investigate the evolution of lung function during the first 6 years of life after a clinical diagnosis of cystic fibrosis (CF) in infancy in children with CF and in healthy control subjects. METHODS: The raised volume technique was used during infancy and incentive spirometry during the preschool years. MEASUREMENTS AND MAIN RESULTS: Forty-eight children with CF and 33 healthy control subjects had up to seven (median, 3) measurements. Over these early years, the diagnosis of CF itself accounted for a significant mean reduction of 7.5% (95% confidence interval, 0.9 - 13.6%) in FEV(0.75) and 15.1% (95% confidence interval, 3.6 - 25.3%) in FEF(25-75). Wheeze on auscultation, recent cough, and Pseudomonas aeruginosa (PsA) infection (even if apparently effectively treated) were all independently associated with further reductions in lung function. Premorbid lung function did not predict infection with PsA. CONCLUSIONS: This is the first study to describe physiologic measurements from infancy through the preschool years in subjects with CF and healthy control subjects, the understanding of which is critical for future intervention trials. Airflow obstruction in uncomplicated CF persists through the preschool years despite treatment, with PsA acquisition being associated with further deterioration in lung function, even when apparently eradicated. This suggests that new therapies are needed to treat the airflow obstruction of uncomplicated CF, and rigorous strategies to prevent PsA acquisition. PMID- 18403722 TI - Maternal food consumption during pregnancy and the longitudinal development of childhood asthma. AB - RATIONALE: Maternal diet during pregnancy has the potential to affect airway development and to promote T-helper-2-cell responses during fetal life. This might increase the risk of developing childhood asthma or allergy. OBJECTIVES: We investigated the influence of maternal food consumption during pregnancy on childhood asthma outcomes from 1 to 8 years of age. METHODS: A birth cohort study consisting of a baseline of 4,146 pregnant women (1,327 atopic and 2,819 nonatopic). These women were asked about their frequency of consumption of fruit, vegetables, fish, egg, milk, milk products, nuts, and nut products during the last month. Their children were followed until 8 years of age. Longitudinal analyses were conducted to assess associations between maternal diet during pregnancy and childhood asthma outcomes over 8 years. MEASUREMENTS AND MAIN RESULTS: Complete data were obtained for 2,832 children. There were no associations between maternal vegetable, fish, egg, milk or milk products, and nut consumption and longitudinal childhood outcomes. Daily consumption of nut products increased the risk of childhood wheeze (odds ratio [OR] daily versus rare consumption, 1.42; 95% confidence interval [95% CI], 1.06-1.89), dyspnea (OR, 1.58; 95% CI, 1.16-2.15), steroid use (OR, 1.62; 95% CI, 1.06-2.46), and asthma symptoms (OR, 1.47; 95% CI, 1.08-1.99). CONCLUSIONS: Results of this study indicate an increased risk of daily versus rare consumption of nut products during pregnancy on childhood asthma outcomes. These findings need to be replicated by other studies before dietary advice can be given to pregnant women. PMID- 18403723 TI - Lung inflammation induced by lipoteichoic acid or lipopolysaccharide in humans. AB - RATIONALE: Recognition of pathogen-associated molecular patterns by Toll-like receptors (TLRs) is considered to be important for an appropriate immune response against pathogens that enter the lower airways. OBJECTIVES: We studied the effects of two different TLR agonists relevant for respiratory infections in the human lung: lipoteichoic acid (LTA; TLR2 agonist, component of gram-positive bacteria) and lipopolysaccharide (LPS; TLR4-agonist, component of gram-negative bacteria). METHODS: Fifteen healthy subjects were given LPS or LTA: by bronchoscope, sterile saline was instilled into a lung segment followed by instillation of LTA or LPS into the contralateral lung. After 6 hours, a bronchoalveolar lavage was performed and inflammatory parameters were determined. Isolated RNA from purified alveolar macrophages was analyzed by multiplex ligation-dependent probe amplification. In addition, spontaneous cytokine release by alveolar macrophages was measured. MEASUREMENTS AND MAIN RESULTS: Marked differences were detected between LTA- and LPS-induced lung inflammation. Whereas both elicited neutrophil recruitment, only LPS instillation was associated with activation of neutrophils (CD11b surface expression, degranulation product levels) and consistent rises of chemo-/cytokine levels. Moreover, LPS but not LTA activated alveolar macrophages, as reflected by enhanced expression of 10 different mRNAs encoding proinflammatory mediators and increased spontaneous cytokine release upon incubation ex vivo. Remarkably, only LTA induced C5a release. CONCLUSIONS: This is the first study to report the in vivo effects of LTA in men and to compare inflammation induced by LTA and LPS in the human lung. Our data suggest that stimulation of TLR2 or TLR4 results in differential pulmonary inflammation, which may be of relevance for understanding pathogenic mechanisms at play during gram-positive and gram-negative respiratory tract infection. PMID- 18403724 TI - Coexpression of CLA-1 and human PDZK1 in murine liver modulates HDL cholesterol metabolism. AB - OBJECTIVE: In rodents scavenger receptor class B type I (SR-BI) is a key molecule for selective uptake of cholesteryl ester from high-density lipoprotein (HDL). This study was aimed to clarify the role of the human SR-BI/CD36 and LIMP-II Analogues-1 (CLA-1) as a molecular target of selective uptake of cholesteryl ester from HDL in vivo. METHODS AND RESULTS: To clarify the function and regulation of CLA-1 in vivo we produced CLA-1 BAC transgenic mice. In spite of abundant hepatic RNA expression of CLA-1, CLA-1 BAC transgenic mice had no significant effect on mouse HDL cholesterol. Although coexpression of a human scaffolding protein PDZK1 along with CLA-1 enhanced hepatic CLA-1 expression, it did not affect mouse HDL cholesterol levels, either. However, in the presence of human apoA-1, HDL cholesterol level and size were significantly reduced in CLA-1 transgenic mice, and its reduction was more pronounced in CLA-1/human PDZK1 double transgenic mouse. CONCLUSIONS: We established a mouse model to study human reverse cholesterol transport by expressing CLA-1, human PDZK1, and human apoA-I gene. Our results imply that enhancing CLA-1 expression by human PDZK1 in the liver can modulate HDL cholesterol metabolism and possibly enhance reverse cholesterol transport to prevent the progression of atherosclerosis in human. PMID- 18403725 TI - Ultrasonic microbubble destruction stimulates therapeutic arteriogenesis via the CD18-dependent recruitment of bone marrow-derived cells. AB - OBJECTIVE: We have previously shown that, under certain conditions, ultrasonic microbubble destruction creates arteriogenesis and angiogenesis in skeletal muscle. Here, we tested whether this neovascularization response enhances hyperemia in a rat model of arterial insufficiency and is dependent on the recruitment of bone marrow-derived cells (BMDCs) to treated tissues via a beta2 integrin (CD18)-dependent mechanism. METHODS AND RESULTS: Sprague-Dawley rats, C57BL/6 wild-type mice, and C57BL/6 chimeric mice engrafted with BMDCs from either GFP+ or CD18-/- mice received bilateral femoral artery ligations. Microbubbles (MBs) were intravenously injected, and one gracilis muscle was exposed to pulsed 1 MHz ultrasound (US). Rat hindlimbs exhibited significant increases in adenosine-induced hyperemia and arteriogenesis compared to contralateral controls at 14 and 28 days posttreatment. US-MB-treated wild-type C57BL/6 mice exhibited significant arteriogenesis, angiogenesis, and CD11b+ monocyte recruitment; however, these responses were all completely blocked in CD18-/- chimeric mice. The number of BMDCs increased in US-MB-treated muscles of GFP+ chimeric mice; however, GFP+ BMDCs did not incorporate into microvessels as vascular cells. CONCLUSIONS: In skeletal muscle affected by arterial occlusion, arteriogenesis and hyperemia can be significantly enhanced by ultrasonic MB destruction. This response depends on the recruitment, but not vascular incorporation, of BMDCs via a CD18-dependent mechanism. PMID- 18403726 TI - Overexpression of ACE2 enhances plaque stability in a rabbit model of atherosclerosis. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that ACE2 overexpression may enhance atherosclerotic plaque stability by antagonizing ACE activity and converting angiotensin II to angiotensin 1-7. METHODS AND RESULTS: Atherosclerotic plaques were induced in the abdominal aorta of 114 rabbits by endothelial injury and atherogenic diet. Gene therapy was performed in group A at week 4 and in group B at week 12, respectively. Each group of rabbits were randomly divided into 3 subgroups which received, respectively, a recombinant ACE2 expressing vector (AdACE2), a control vector AdEGFP and AdACE2+A779, an antagonist of angiotensin 1-7 receptor. Local ACE2 overexpression attenuated the progression of lesions from week 4 to week 8, but not progression of plaque size from week 12 to week 16. In group B rabbits, local ACE2 overexpression resulted in stable plaque compositions, ie, fewer macrophages, less lipid deposition and more collagen contents, higher plaque stability scores, decreased angiotensin II levels, and increased angiotensin 1-7 levels in plaque tissues in the AdACE2 subgroup compared with those in the AdEGFP subgroup. CONCLUSIONS: Overexpression of ACE2 results in stabilized atherosclerotic plaques and the mechanism is probably the conversion of vasoconstrictive angiotensin II to vessel protective angiotensin 1-7. PMID- 18403727 TI - Augmentation of vascular remodeling by uncoupled endothelial nitric oxide synthase in a mouse model of diabetes mellitus. AB - OBJECTIVE: Diabetes mellitus is associated with increased oxidative stress, which induces oxidation of tetrahydrobiopterin (BH4) in vessel wall. Without enough BH4, eNOS is uncoupled to L-arginine and produces superoxide rather than NO. We examined the role of uncoupled eNOS in vascular remodeling in diabetes. METHODS AND RESULTS: Diabetes mellitus was produced by streptozotocin in C57BL/6J mice. Under stable hyperglycemia, the common carotid artery was ligated, and neointimal formation was examined 4 weeks later. In diabetic mice, the neointimal area was dramatically augmented. This augmentation was associated with increased aortic superoxide formation, reduced aortic BH4/dihydrobiopterin (BH2) ratio, and decreased plasma nitrite and nitrate (NOx) levels compared with nondiabetic mice. Chronic BH4 treatment (10 mg/kg/d) reduced the neointimal area in association with suppressed superoxide production and inflammatory changes in vessels. BH4/BH2 ratio in vessel wall was preserved, and plasma NOx levels increased. Furthermore, in the presence of diabetes, overexpression of bovine eNOS resulted in augmentation of neointimal area, accompanied by increased superoxide production in the endothelium. CONCLUSIONS: In diabetes, increased oxidative stress by uncoupled NOSs, particularly eNOS, causes augmentation of vascular remodeling. These findings indicate restoration of eNOS coupling has an atheroprotective benefit in diabetes. PMID- 18403728 TI - High-sensitivity C-reactive protein and coronary heart disease in a general population of Japanese: the Hisayama study. AB - OBJECTIVE: The purpose of this study was to investigate the effects of high sensitivity C-reactive protein (hs-CRP) on the risks of coronary heart disease (CHD) in a general population of Japanese. METHODS AND RESULTS: The Hisayama study is a population-based prospective cohort study. A total of 2589 participants aged 40 years or older were followed up for 14 years. Outcomes are incident CHD (myocardial infarction, coronary revascularization, and sudden cardiac death). The median hs-CRP level was 0.43 mg/L at baseline. During the follow-up period, 129 coronary events were observed. Age- and sex-adjusted annual incidence rates of CHD rose progressively with higher hs-CRP levels: 1.6, 3.3, 4.5, and 7.4 per 1000 person-years for quartile groups defined by hs-CRP levels of <0.21, 0.21 to 0.43, 0.44 to 1.02, and >1.02 mg/L, respectively (P<0.0001 for trend). The risk of CHD in the highest quartile group was 2.98-fold (95% CI, 1.53 to 5.82) higher than that in the lowest group even after controlling for other cardiovascular risk factors. CONCLUSIONS: hs-CRP levels were clearly associated with future CHD events in a general population of Japanese. In Japanese populations, the hs-CRP cut-off point for high-risk of future development of CHD is likely to be >1.0 mg/L, which is much lower than that for Western populations. PMID- 18403729 TI - Modulation of endothelial cell KCa3.1 channels during endothelium-derived hyperpolarizing factor signaling in mesenteric resistance arteries. AB - Arterial hyperpolarization to acetylcholine (ACh) reflects coactivation of K(Ca)3.1 (IK(Ca)) channels and K(Ca)2.3 (SK(Ca)) channels in the endothelium that transfers through myoendothelial gap junctions and diffusible factor(s) to affect smooth muscle relaxation (endothelium-derived hyperpolarizing factor [EDHF] response). However, ACh can differentially activate K(Ca)3.1 and K(Ca)2.3 channels, and we investigated the mechanisms responsible in rat mesenteric arteries. K(Ca)3.1 channel input to EDHF hyperpolarization was enhanced by reducing external [Ca(2+)](o) but blocked either with forskolin to activate protein kinase A or by limiting smooth muscle [Ca(2+)](i) increases stimulated by phenylephrine depolarization. Imaging [Ca(2+)](i) within the endothelial cell projections forming myoendothelial gap junctions revealed increases in cytoplasmic [Ca(2+)](i) during endothelial stimulation with ACh that were unaffected by simultaneous increases in muscle [Ca(2+)](i) evoked by phenylephrine. If gap junctions were uncoupled, K(Ca)3.1 channels became the predominant input to EDHF hyperpolarization, and relaxation was inhibited with ouabain, implicating a crucial link through Na(+)/K(+)-ATPase. There was no evidence for an equivalent link through K(Ca)2.3 channels nor between these channels and the putative EDHF pathway involving natriuretic peptide receptor-C. Reconstruction of confocal z-stack images from pressurized arteries revealed K(Ca)2.3 immunostain at endothelial cell borders, including endothelial cell projections, whereas K(Ca)3.1 channels and Na(+)/K(+)-ATPase alpha(2)/alpha(3) subunits were highly concentrated in endothelial cell projections and adjacent to myoendothelial gap junctions. Thus, extracellular [Ca(2+)](o) appears to modify K(Ca)3.1 channel activity through a protein kinase A-dependent mechanism independent of changes in endothelial [Ca(2+)](i). The resulting hyperpolarization links to arterial relaxation largely through Na(+)/K(+)-ATPase, possibly reflecting K(+) acting as an EDHF. In contrast, K(Ca)2.3 hyperpolarization appears mainly to affect relaxation through myoendothelial gap junctions. Overall, these data suggest that K(+) and myoendothelial coupling evoke EDHF-mediated relaxation through distinct, definable pathways. PMID- 18403730 TI - S100A8 and S100A9 mediate endotoxin-induced cardiomyocyte dysfunction via the receptor for advanced glycation end products. AB - Cardiovascular dysfunction as a result of sepsis is the leading cause of death in the critically ill. Cardiomyocytes respond to infectious pathogens with a Toll like receptor-initiated proinflammatory response in conjunction with a decrease in contractility, although the downstream events linking Toll-like receptor activation and reduced cardiac contractility remain to be elucidated. Using microarray analysis of cardiac tissue exposed to systemic lipopolysaccharide (LPS), we discovered that 2 small calcium-regulating proteins (S100A8 and S100A9) are highly upregulated. HL-1 cardiomyocytes, isolated primary cardiomyocytes, and live mice were exposed to LPS, whereas beating HL-1 cells had S100A8 and S100A9 overexpressed and their calcium flux quantified. Using in vivo microbubble technology, we delivered S100A8 and S100A9 to normal mouse hearts; using the same technology, we inhibited S100A9 production in mouse hearts and subsequently exposed them to LPS. Coimmunoprecipitation of S100A8 and S100A9 identified interaction with RAGE (the receptor for advanced glycation end products), the cardiac function and postreceptor signaling of which were investigated. HL-1 cardiomyocytes, isolated primary cardiomyocytes, and whole hearts exposed to LPS have large increases in S100A8 and S100A9. Cardiac overexpression of S100A8 and S100A9 led to a RAGE-dependent decrease in calcium flux and, in the intact mouse, to a decreased cardiac ejection fraction, whereas knockdown of S100A9 attenuated LPS-induced cardiac dysfunction. Cardiomyocytes exposed to LPS express S100A8 and S100A9, leading to a RAGE-mediated decrease in cardiomyocyte contractility. This finding provides a novel mechanistic link between circulating pathogen-associated molecular products and subsequent cardiac dysfunction. PMID- 18403731 TI - Dragging along: the glycocalyx and vascular endothelial cell mechanotransduction. PMID- 18403732 TI - Pitx2: a challenging teenager. PMID- 18403733 TI - Critical cap thickness and rupture in symptomatic carotid plaques: the oxford plaque study. AB - BACKGROUND AND PURPOSE: Advances in carotid plaque imaging could allow quantification of fibrous cap thickness in vivo. While a cap thickness <65 microm is the accepted definition of rupture-prone plaque in the coronary circulation, the threshold value for carotid plaques is unknown. METHODS: We made detailed histological assessments of 526 carotid plaques from consecutive patients undergoing endarterectomy for symptomatic carotid stenosis. The thickness of the fibrous cap at the thinnest and most representative part was measured. RESULTS: Cap thickness could be measured reliably in 428 (81%) plaques. In the ruptured plaques (n=257), the median representative cap thickness was 300 microm (IQR 200 to 500 microm) and the median minimum cap thickness was 150 microm (80 to 210 microm; mean=181 microm), which is much greater than the mean cap thickness of 23 microm at the point of rupture that has been reported for coronary plaques. For nonruptured plaques, the median cap thickness values were 500 microm (300 to 700 microm) and 250 microm (180 to 400 microm), respectively. The optimum cut-offs for discriminating between ruptured and nonruptured plaques were a minimum cap thickness <200 microm (OR 5.00, 3.26 to 7.65, P<0.001), a representative cap thickness <500 microm (OR 3.38, 2.25 to 5.08, P<0.001), or a combination of both (OR 5.11, 3.19 to 8.19, P<0.001). Minimum and representative cap thickness were only modestly correlated (r(2)=0.30) and were both independently associated with cap rupture. CONCLUSIONS: Critical cap thickness is greater in carotid plaques than coronary plaques. Minimum and representative cap thicknesses were both independently associated with cap rupture. A combination of minimum cap thickness <200 microm and a representative cap thickness <500 microm identified ruptured plaques most reliably. Prospective imaging studies are required to establish whether these cut points predict clinical events in patients with asymptomatic carotid stenosis. PMID- 18403734 TI - Polymorphisms in platelet glycoprotein 1balpha and factor VII and risk of ischemic stroke: a meta-analysis. AB - BACKGROUND AND PURPOSE: Platelets and components of the coagulation cascade are known to be instrumental in the pathogenesis of arterial occlusive disorders. The aim of this meta-analysis is to test the hypothesis that genetic variation in the platelet glycoprotein 1balpha and Factor VII genes influence the occurrence of ischemic stroke. All genetic association studies that examined the R353Q (rs6046) polymorphism of the Factor VII gene and 2 polymorphisms of the platelet glycoprotein (1balpha) gene (Thr/Met rs6065 and Kozak sequence -5 C/T rs2243093) in relation to ischemic stroke were examined. METHODS: Electronic databases Embase, Medline, and HuGEnet were searched for all years up until June 2006 for all studies that evaluated any of these candidate genes and stroke. RESULTS: Pooled ORs were calculated with 95% CIs using both fixed and random effects models. Meta-analysis for Factor VII (R353Q) did not detect any effect on ischemic stroke risk. Further estimation resulted in pooled OR(1) QQ versus RR=0.9 (95% CI: 0.4 to 1.9) and pooled OR(2) for RQ versus RR=0.9 (95% CI: 0.6 to 1.4). These results were robust and homogeneous. Pooling ORs for the platelet glycoprotein 1balpha Kozak variant -5 T/C polymorphism showed extreme heterogeneity with differing effect directions across studies. Fisher's method of pooling was therefore used to calculate a combined probability value, which was highly significant (P<0.001). The pooled OR for platelet glycoprotein 1balpha Met/Met v Thr/Thr was 1.0 to 2.0, depending on the sensitivity analyses, and for Thr/Met versus Thr/Thr, the pooled OR was between 1.3 and 1.4. These results were consistent, reasonably robust, and implied a dominant genetic effect. CONCLUSIONS: This analysis provides strong evidence that the Factor VII R353Q gene polymorphism is not associated with ischemic stroke, that the Thr/Met polymorphism of GP1balpha is associated with ischemic stroke in a dominant genetic model, and that the Kozak sequence polymorphism of GP1balpha may be close to another causative locus that is associated with ischemic stroke. PMID- 18403735 TI - Meta-analysis of the efficacy of granulocyte-colony stimulating factor in animal models of focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: Recent reports have described the efficacy of the hematopoietic growth factor granulocyte-colony stimulating factor (G-CSF) in animal stroke models. Early clinical multicenter trials evaluating the effect of G-CSF in acute stroke and pilot clinical trials for the subacute phase are ongoing. To guide further development, a meta-analysis was performed to assess the effects of G-CSF on infarct size and sensorimotor deficits. METHODS: Using electronic and manual searches of the literature, we identified studies describing the efficacy of G-CSF in animal models of focal cerebral ischemia. Two reviewers independently selected studies and extracted data on study quality, G CSF doses, time of administration, and outcome measured as infarct volume and/or sensorimotor deficit. Data from all studies were pooled by meta-regression analyses. RESULTS: Thirteen studies including 277 animals for infarct size calculation and 258 animals for assessment of sensorimotor deficit met the criteria for inclusion. Overall efficacy of G-CSF regarding infarct size reduction was 42%. Meta-regression analysis revealed a 0.8% (P<0.0001) decrease in infarct size per 1-mug/kg increase in G-CSF dose when applied within the first 6 hours and a 2.1% (P<0.0001) decrease when applied later than 6 hours after induction of ischemia with a significant (P=0.0004) greater infarct size reduction after delayed treatment. Sensorimotor deficits categorized into 3 subgroups improved between 24% and 40%. CONCLUSIONS: Our findings consolidate G CSF as a drug that both reduces infarct size and enhances functional recovery. These effects are presumably dose dependent. In contrast to most other neuroprotectants, a beneficial outcome may also be achieved when treatment is delayed. PMID- 18403736 TI - DP-b99, a membrane-activated metal ion chelator, as neuroprotective therapy in ischemic stroke. AB - BACKGROUND AND PURPOSE: DP-b99 is a chelator of zinc and calcium ions that acts selectively within cell membranes and has neuroprotective properties in animal models of stroke. We present the results of a multicenter, double-blind, placebo controlled, randomized trial to assess the safety and potential protective effects of DP-b99 in acute ischemic stroke. METHODS: One hundred and fifty stroke patients with signs of cortical involvement and a National Institutes of Health Stroke Scale (NIHSS) score of 7 to 20 received a 4-day course of intravenous 1 mg/kg per day DP-b99 or placebo within 1 to 9 hours of stroke onset. Treatment with recombinant tissue plasminogen activator was not allowed. RESULTS: No major differences in mortality rate, causes of death, adverse events, safety laboratory tests, and ECG parameters were found between the 2 groups. The baseline NIHSS score of the 72 DP-b99- and 75 placebo-treated patients in the intent-to-treat cohort was (mean+/-SD) 12.2+/-4.0 and 12.6+/-3.3, respectively; the time to needle (mean+/-SD) was 6:36+/-1:47 and 6:28+/-1:33 hours, respectively; and the age (mean+/-SD) was 73.3+/-9.9 and 72.0+/-9.6 years, respectively. The 90-day median change from baseline (the primary end point) was -6.0 and -5.0 NIHSS points in the DP-b99 and placebo groups, respectively (nonsignificant). At 90 days, there was a significantly better outcome in the DP-b99 group compared with the placebo group (modified Rankin scale score of 0, 1, or same as prestroke): 30.6% and 16.0%, respectively (P=0.05). The recovery rate was unaffected by the time to needle. Further analyses indicated that the 90-day median change from baseline in patients with an entry NIHSS score of 10 to 16 was 8.0 and 5.0 points in the DP-b99 and placebo groups, respectively (P=0.03). CONCLUSIONS: In this small-scale study, the primary end point of change in NIHSS score from baseline to 90 days was not met. However, secondary end points demonstrated a significantly improved 90-day recovery rate with treatment with DP-b99 when compared with placebo. In addition, in patients with baseline NIHSS scores of 10 to 16, a significant post hoc change in NIHSS score from baseline to day 90 was observed. No major safety problems were identified. These findings need to be confirmed with a larger prospective study of strokes involving the cortex. PMID- 18403738 TI - Predicting long-term outcome after acute ischemic stroke: a simple index works in patients from controlled clinical trials. AB - BACKGROUND AND PURPOSE: An early and reliable prognosis for recovery in stroke patients is important for initiation of individual treatment and for informing patients and relatives. We recently developed and validated models for predicting survival and functional independence within 3 months after acute stroke, based on age and the National Institutes of Health Stroke Scale score assessed within 6 hours after stroke. Herein we demonstrate the applicability of our models in an independent sample of patients from controlled clinical trials. METHODS: The prognostic models were used to predict survival and functional recovery in 5419 patients from the Virtual International Stroke Trials Archive (VISTA). Furthermore, we tried to improve the accuracy by adapting intercepts and estimating new model parameters. RESULTS: The original models were able to correctly classify 70.4% (survival) and 72.9% (functional recovery) of patients. Because the prediction was slightly pessimistic for patients in the controlled trials, adapting the intercept improved the accuracy to 74.8% (survival) and 74.0% (functional recovery). Novel estimation of parameters, however, yielded no relevant further improvement. CONCLUSIONS: For acute ischemic stroke patients included in controlled trials, our easy-to-apply prognostic models based on age and National Institutes of Health Stroke Scale score correctly predicted survival and functional recovery after 3 months. Furthermore, a simple adaptation helps to adjust for a different prognosis and is recommended if a large data set is available. PMID- 18403737 TI - Effects of neuroglobin overexpression on acute brain injury and long-term outcomes after focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: Emerging data suggest that neuroglobin (Ngb) may protect against hypoxic/ischemic neuronal insults. However, the underlying mechanisms in vivo and implications for long-term outcomes are still not well understood. METHODS: Using our newly created Ngb overexpressing transgenic (Ngb-Tg) mice, we measured brain infarction on day 1 and day 14 after transient focal cerebral ischemia and performed neurobehavioral assessments in sensorimotor deficits on days 1, 3, 7, and 14. To test the hypothesis that Ngb may play a role in reducing oxidative stress after stroke, intracellular malondialdehyde levels were measured and compared in Ngb-Tg and wild-type mice. RESULTS: Increased Ngb mRNA and protein levels were identified in Ngb-Tg brains. Malondialdehyde levels in ischemic hemispheres of Ngb-Tg were significantly reduced compared with wild-type controls at 8 hours and 22 hours after transient focal cerebral ischemia. Compared with wild-type controls, brain infarction volumes 1 day and 14 days after transient focal cerebral ischemia were significantly reduced in Ngb-Tg mice. However, there were no significant improvements in sensorimotor deficits for up to 14 days after stroke in Ngb-Tg mice compared with wild-type controls. CONCLUSIONS: Ngb reduces tissue infarction and markers of oxidative stress after stroke. Tissue protection by overexpressing Ngb can be sustained for up to 2 weeks. PMID- 18403739 TI - Assessment of intracranial collateral flow by using dynamic arterial spin labeling MRA and transcranial color-coded duplex ultrasound. AB - BACKGROUND AND PURPOSE: To evaluate the potential of a new dynamic MRA sequence (DynAngio) based on arterial spin labeling for the assessment of intracranial collateral flow. METHODS: Twelve patients with unilateral internal carotid artery obstruction were investigated. Different patterns of collateral flow were compared between DynAngio, transcranial color-coded duplex ultrasound, and time of-flight MRA. RESULTS: There was a good agreement between the methods, with sensitivities between 80% and 90%. Small collateral vessels were detected more frequently with DynAngio compared to time-of-flight MRA. CONCLUSIONS: DynAngio provides anatomic and similar to transcranial color-coded duplex ultrasound functional information on collateral flow for the assessment of intracranial hemodynamics. PMID- 18403740 TI - Characterization of cortical microvascularization in adult moyamoya disease. AB - BACKGROUND AND PURPOSE: Increased cortical microvascularization has been proposed to be a Moyamoya disease (MMD)-specific characteristic. It was the aim of our study to characterize the anatomic pattern and microhemodynamics of cortical microvascularization in MMD. METHODS: Intraoperative indocyanine green videoangiography was performed in 16 adult MMD patients, 15 patients with atherosclerotic cerebrovascular disease (ACVD), and 10 control patients. Cortical microvascularization and microvascular hemodynamics were categorized and analyzed according to anatomic and functional indocyanine green angiographic aspects. Anatomic analysis included microvascular density, microvascular diameter, and microvascular surface per analyzed area. Microhemodynamic analysis included microvascular transit time, arterial microvascular transit time, and venous microvascular transit time. RESULTS: Microvascular density and diameter were significantly increased in MMD patients (1.8+/-0.2 mm/mm(2) and 0.24+/-0.03 mm, respectively) compared with those in ACVD patients (1.5+/-0.2 mm/mm(2) and 0.20+/ 0.02 mm, respectively) and controls (1.5+/-0.1 mm/mm(2) and 0.19+/-0.03 mm, respectively). This resulted in significantly increased microvascular surface per analyzed area in MMD (67+/-13%) vs ACVD patients (47+/-7%) and controls (45+/ 6%). Anatomic changes were paralleled by significantly increased microvascular and arterial microvascular transit times in MMD patients (11.55+/-3.50 and 6.79+/ 2.96 seconds, respectively) compared with those in ACVD patients (8.13+/-1.78 and 4.34+/-1.30 seconds, respectively) and controls (8.04+/-2.16 and 4.50+/-1.87 seconds, respectively). CONCLUSIONS: Cortical microvascularization in MMD is characterized by significantly increased microvascular density and microvascular diameter, leading to increased microvascular surface. These anatomic alterations are accompanied by prolonged microvascular hemodynamics. These observations might represent an MMD-specific compensation mechanism for impaired cerebral blood flow. PMID- 18403741 TI - Fas system activation in perihematomal areas after spontaneous intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Apoptosis has been implicated as the prominent form of cell death in the brain perihematomal region in animal models and in autopsy or postsurgical human studies. Both the Fas system and caspase activation play a central role in apoptotic pathways. The aims of this study were to investigate soluble Fas (s-Fas) plasma levels after acute intracerebral hemorrhage (ICH), to determine its influence on clinical and radiologic features, and to assess Fas receptor and Fas ligand (Fas-L) protein expression in human ICH brain tissue. METHODS: s-Fas plasma levels were determined on admission in 78 consecutive ICH patients and serially in a subgroup of 21 of them, at the time of neurologic assessment, by means of ELISA. ICH and perihematomal edema volumes were determined at baseline and on follow-up computed tomography scans, and ICH and perihematomal edema growth was calculated. The presence of Fas receptor and Fas-L was assessed in different brain tissue samples by immunoblotting from 6 deceased ICH patients and from 2 control subjects. RESULTS: Mortality reached 20.5% of patients at the third month, and 48% of survivors had an unfavorable outcome (modified Rankin Scale score >/=3). The baseline s-Fas level in ICH patients was significantly lower than in healthy controls [160 (160-245) vs 269 (230-332) pg/mL, P<0.001], returning to normal values by 24 hours (P<0.05 for all determinations). Regarding radiologic features, the baseline s-Fas value was found to be inversely correlated to perihematomal edema growth at follow-up (r= 0.33, P=0.041). Finally, Fas-L content was highest in the perihematomal area compared with contralateral and remote ipsilateral areas in ICH patient and control samples. CONCLUSIONS: A decreased plasma s-Fas level together with an increased Fas-L amount in perihematomal brain tissue suggest Fas-mediated apoptosis involvement in this disease. PMID- 18403742 TI - Training of reaching in stroke survivors with severe and chronic upper limb paresis using a novel nonrobotic device: a randomized clinical trial. AB - BACKGROUND AND PURPOSE: Severe upper limb paresis is a major contributor to disability after stroke. This study investigated the efficacy of a new nonrobotic training device, the Sensorimotor Active Rehabilitation Training (SMART) Arm, that was used with or without electromyography-triggered electrical stimulation of triceps brachii to augment elbow extension, permitting stroke survivors with severe paresis to practice a constrained reaching task. METHODS: A single-blind, randomized clinical trial was conducted with 42 stroke survivors with severe and chronic paresis. Thirty-three participants completed the study, of whom 10 received training using the SMART Arm with electromyography-triggered electrical stimulation, 13 received training using the SMART Arm alone, and 10 received no intervention (control). Training consisted of 12 1-hour sessions over 4 weeks. The primary outcome measure was "upper arm function," item 6 of the Motor Assessment Scale. Secondary outcome measures included impairment measures; triceps muscle strength, reaching force, modified Ashworth scale; and activity measures: reaching distance and Motor Assessment Scale. Assessments were administered before (0 weeks) and after training (4 weeks) and at 2 months follow up (12 weeks). RESULTS: Both SMART Arm groups demonstrated significant improvements in all impairment and activity measures after training and at follow up. There was no significant difference between these 2 groups. There was no change in the control group. CONCLUSIONS: Our findings indicate that training of reaching using the SMART Arm can reduce impairment and improve activity in stroke survivors with severe and chronic upper limb paresis, highlighting the benefits of intensive task-oriented practice, even in the context of severe paresis. PMID- 18403743 TI - Acute stroke imaging research roadmap. AB - The recent "Advanced Neuroimaging for Acute Stroke Treatment" meeting on September 7 and 8, 2007 in Washington DC, brought together stroke neurologists, neuroradiologists, emergency physicians, neuroimaging research scientists, members of the National Institute of Neurological Disorders and Stroke (NINDS), the National Institute of Biomedical Imaging and Bioengineering (NIBIB), industry representatives, and members of the US Food and Drug Administration (FDA) to discuss the role of advanced neuroimaging in acute stroke treatment. The goals of the meeting were to assess state-of-the-art practice in terms of acute stroke imaging research and to propose specific recommendations regarding: (1) the standardization of perfusion and penumbral imaging techniques, (2) the validation of the accuracy and clinical utility of imaging markers of the ischemic penumbra, (3) the validation of imaging biomarkers relevant to clinical outcomes, and (4) the creation of a central repository to achieve these goals. The present article summarizes these recommendations and examines practical steps to achieve them. PMID- 18403744 TI - Predictions models in acute stroke: potential uses and limitations. PMID- 18403745 TI - Are we ready to avert suicide in intracerebral hemorrhage? PMID- 18403747 TI - Stop the sores: the making and evaluation of a successful social marketing campaign. AB - Los Angeles County has experienced a rapid increase in early syphilis among men who have sex with men (MSM) in recent years, with the number of cases rising from 126 in 2000 to 809 in 2005. As part of the public health response to this outbreak, a sustained social marketing campaign was launched in 2002, with the objectives of increasing syphilis testing, knowledge, and awareness among MSM in Los Angeles. This campaign, as implemented, exemplified key principles of social marketing, including market research, audience segmentation, and branding. A cross-sectional study conducted in 2004 to evaluate the campaign found that those MSM who were aware of the campaign were nearly twice as likely to have tested for syphilis in the past 6 months as those MSM who were not aware of the campaign. Those MSM who were aware of the campaign also had more syphilis awareness and knowledge in key areas. PMID- 18403746 TI - Effects of action observation on physical training after stroke. AB - BACKGROUND AND PURPOSE: In healthy humans, observation of another individual performing a motor training task (action observation [AO]) facilitates, in the observer, the effects of physical training (PT) on motor memory formation. It is not known whether this facilitatory process, of potential value for neurorehabilitation, occurs after stroke. METHODS: Eight chronic stroke patients completed this crossover-randomized investigation. A transcranial magnetic stimulation protocol that tests formation of motor memories was used to determine the effects of PT alone and in combination with AO in 2 different forms: congruent (PT+AO(congruent)) and incongruent (PT+AO(incongruent)) to the practiced task. RESULTS: The magnitude of motor memory formation was larger with PT+AO(congruent) than with PT alone or PT+AO(incongruent). This effect was associated with a differential corticomotor excitability change in the muscles acting as agonist and antagonist of the trained/observed movements. CONCLUSIONS: These results indicate that congruent AO in association with physical training can enhance the effects of motor training after stroke. PMID- 18403748 TI - A descriptive analysis of patient education courses in undergraduate and graduate health education programs. AB - Patient education is a necessary component of quality health care, yet little attention has been given to the preparation of health educators to work in that setting. This study seeks to determine the status of and content in patient education courses offered in professional preparation programs. Results show that 9% of respondents offered a patient education course in their academic unit, whereas 18% indicated that such a course was offered in another unit on campus. It appears there is not agreement between university faculty members and practicing patient educators on what should be taught in such a course. In addition, no significant relationship is found between (a) programs with accreditation or approval and offering a patient education course and (b) programs that prepared students for the Certified Health Education Specialist examination and offering a patient education course. Recommendations are offered for improving the preparation of health educators for the medical care setting. PMID- 18403749 TI - Novel use of patient-derived vignettes to foster self-efficacy in an asthma self management workbook. AB - This article describes the development of an interactive self-management workbook to increase asthma knowledge and self-efficacy. Its format encourages readers to write in their own personal experiences and perspectives. The knowledge component was developed from standard materials and organized into thematic chapters. The self-efficacy component was developed from interviews with patients in the outpatient setting, emergency room, and hospital; it consists of three sections explaining why confidence is necessary to be an effective self-manager and offers techniques to increase confidence. The component's cornerstone is vignettes of patients describing what triggers their asthma and what behaviors they use to successfully manage it. This novel approach to fostering self-efficacy allows patients to see themselves in others' success stories. Although not a substitute for self-management courses, the workbook promotes self-management by offering a practical and readily available option to in-person instruction and peer contact, which are accessible to only a small number of patients. PMID- 18403750 TI - Totipotency/pluripotency and patentability. AB - In their article entitled " COMMENTARY: Is totipotency of a human cell a sufficient reason to exclude its patentability under the European law" (Stem Cells 2007;25:3026-3028), K.T. Vrtovec and B. Vrtovec conclude that arguments based on differentiation potential should not be an obstacle to patenting human embryonic stem cells (and related cells referred to as totipotent or pluripotent). While concentrating on formal legal aspects, however, these authors fail to consider a major biological and ethical argument already found in the literature, namely that an obstacle to patenting is to be seen in the potential of cells (e.g., of embryonic stem cell lines), if this potential allows (re)constitution of an embryo when tetraploid complementation is performed. PMID- 18403751 TI - Urokinase plasminogen activator and urokinase plasminogen activator receptor mediate human stem cell tropism to malignant solid tumors. AB - Human neural and mesenchymal stem cells have been identified for cell-based therapies in regenerative medicine and as vehicles for delivering therapeutic agents to areas of injury and tumors. However, the signals required for homing and recruitment of stem cells to these sites are not well understood. Urokinase plasminogen activator (uPA) and urokinase plasminogen activator receptor (uPAR) are involved in chemotaxis and cell guidance during normal development and are upregulated in invasive tumors. Here we provided evidence that activation of uPA and uPAR in malignant solid tumors (brain, lung, prostate, and breast) augments neural and mesenchymal stem cell tropism. Expression levels of uPAR on human solid tumor cell lines correlated with levels of uPA and soluble uPAR in tumor cell-conditioned media. Cytokine expression profiles of these tumor-conditioned media were determined by protein arrays. Among 79 cytokines investigated, interleukin (IL)-6, IL-8, and monocyte chemoattractant protein-1 were the most highly expressed cytokines in uPAR-positive tumors. We provided evidence that human recombinant uPA induced stem cell migration, whereas depletion of uPA from PC-3 prostate cancer cell-conditioned medium blocked stem cell migration. Furthermore, retrovirus-mediated overexpression of uPA and uPAR in neuroblastoma (NB1691) cells induced robust migration of stem cells toward NB1691 cell conditioned media, compared with media derived from wild-type NB1691 cells. We conclude that expression of uPA and uPAR in cancer cells underlies a novel mechanism of stem cell tropism to malignant solid tumors, which may be important for development of optimal stem cell-based therapies. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18403752 TI - Polycomb repressive complex 2 is dispensable for maintenance of embryonic stem cell pluripotency. AB - Polycomb repressive complex 2 (PRC2) methylates histone H3 tails at lysine 27 and is essential for embryonic development. The three core components of PRC2, Eed, Ezh2, and Suz12, are also highly expressed in embryonic stem (ES) cells, where they are postulated to repress developmental regulators and thereby prevent differentiation to maintain the pluripotent state. We performed gene expression and chimera analyses on low- and high-passage Eed(null) ES cells to determine whether PRC2 is required for the maintenance of pluripotency. We report here that although developmental regulators are overexpressed in Eed(null) ES cells, both low- and high-passage cells are functionally pluripotent. We hypothesize that they are pluripotent because they maintain expression of critical pluripotency factors. Given that EED is required for stability of EZH2, the catalytic subunit of the complex, these data suggest that PRC2 is not necessary for the maintenance of the pluripotent state in ES cells. We propose a positive-only model of embryonic stem cell maintenance, where positive regulation of pluripotency factors is sufficient to mediate stem cell pluripotency. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18403753 TI - Comprehensive microRNA profiling reveals a unique human embryonic stem cell signature dominated by a single seed sequence. AB - Embryonic stem cells are unique among cultured cells in their ability to self renew and differentiate into a wide diversity of cell types, suggesting that a specific molecular control network underlies these features. Human embryonic stem cells (hESCs) are known to have distinct mRNA expression, global DNA methylation, and chromatin profiles, but the involvement of high-level regulators, such as microRNAs (miRNA), in the hESC-specific molecular network is poorly understood. We report that global miRNA expression profiling of hESCs and a variety of stem cell and differentiated cell types using a novel microarray platform revealed a unique set of miRNAs differentially regulated in hESCs, including numerous miRNAs not previously linked to hESCs. These hESC-associated miRNAs were more likely to be located in large genomic clusters, and less likely to be located in introns of coding genes. hESCs had higher expression of oncogenic miRNAs and lower expression of tumor suppressor miRNAs than the other cell types. Many miRNAs upregulated in hESCs share a common consensus seed sequence, suggesting that there is cooperative regulation of a critical set of target miRNAs. We propose that miRNAs are coordinately controlled in hESCs, and are key regulators of pluripotence and differentiation. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18403754 TI - Platelet-derived growth factor-D overexpression contributes to epithelial mesenchymal transition of PC3 prostate cancer cells. AB - The majority of human malignancies are believed to have epithelial origin, and the progression of cancer is often associated with a transient process named epithelial-mesenchymal transition (EMT). EMT is characterized by the loss of epithelial markers and the gain of mesenchymal markers that are typical of "cancer stem-like cells," which results in increased cell invasion and metastasis in vivo. Therefore, it is important to uncover the mechanistic role of factors that may induce EMT in cancer progression. Studies have shown that platelet derived growth factor (PDGF) signaling contributes to EMT, and more recently, PDGF-D has been shown to regulate cancer cell invasion and angiogenesis. However, the mechanism by which PDGF-D promotes invasion and metastases and whether it is due to the acquisition of EMT phenotype remain elusive. For this study, we established stably transfected PC3 cells expressing high levels of PDGF-D, which resulted in the significant induction of EMT as shown by changes in cellular morphology concomitant with the loss of E-cadherin and zonula occludens-1 and gain of vimentin. We also found activation of mammalian target of rapamycin and nuclear factor-kappaB, as well as Bcl-2 overexpression, in PDGF-D PC3 cells, which was associated with enhanced adhesive and invasive behaviors. More importantly, PDGF-D-overexpressing PC3 cells showed tumor growth in SCID mice much more rapidly than PC3 cells. These results provided a novel mechanism by which PDGF-D promotes EMT, which in turn increases tumor growth, and these results further suggest that PDGF-D could be a novel therapeutic target for the prevention and/or treatment of prostate cancer. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18403755 TI - Direct orthotopic transplantation of fresh surgical specimen preserves CD133+ tumor cells in clinically relevant mouse models of medulloblastoma and glioma. AB - Recent identification of cancer stem cells in medulloblastoma (MB) and high-grade glioma has stimulated an urgent need for animal models that will not only replicate the biology of these tumors, but also preserve their cancer stem cell pool. We hypothesize that direct injection of fresh surgical specimen of MB and high-grade glioma tissues into anatomically equivalent locations in immune deficient mouse brains will facilitate the formation of clinically accurate xenograft tumors by allowing brain tumor stem cells, together with their non-stem tumor and stromal cells, to grow in a microenvironment that is the closest to human brains. Eight of the 14 MBs (57.1%) and two of the three high-grade gliomas (66.7%) in this study developed transplantable (up to 12 passages) xenografts in mouse cerebellum and cerebrum, respectively. These xenografts are patient specific, replicating the histopathologic, immunophenotypic, invasive/metastatic, and major genetic (analyzed with 10K single nucleotide polymorphism array) abnormalities of the original tumors. The xenograft tumor cells have also been successfully cryopreserved for long-term preservation of tumorigenicity, ensuring a sustained supply of the animal models. More importantly, the CD133(+) tumor cells, ranging from 0.2%-10.4%, were preserved in all the xenograft models following repeated orthotopic subtransplantations in vivo. The isolated CD133(+) tumor cells formed neurospheres and displayed multi-lineage differentiation capabilities in vitro. In summary, our study demonstrates that direct orthotopic transplantation of fresh primary tumor cells is a powerful approach in developing novel clinical relevant animal models that can reliably preserve CD133(+) tumor cell pools even during serial in vivo subtransplantations. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 18403756 TI - Impact of myocardial infarct proteins and oscillating pressure on the differentiation of mesenchymal stem cells: effect of acute myocardial infarction on stem cell differentiation. AB - Stem cell transplantation in acute myocardial infarction (AMI) has emerged as a promising therapeutic option. We evaluated the impact of AMI on mesenchymal stem cell (MSC) differentiation into cardiomyocyte lineage. Cord blood-derived human MSCs were exposed to in vitro conditions simulating in vivo environments of the beating heart with acute ischemia, as follows: (a) myocardial proteins or serum obtained from sham-operated rats, and (b) myocardial proteins or serum from AMI rats, with or without application of oscillating pressure. Expression of cardiac specific markers on MSCs was greatly induced by the infarcted myocardial proteins, compared with the normal proteins. It was also induced by application of oscillating pressure to MSCs. Treatment of MSCs with infarcted myocardial proteins and oscillating pressure greatly augmented expression of cardiac specific genes. Such expression was blocked by inhibitor of transforming growth factor beta(1) (TGF-beta(1)) or bone morphogenetic protein-2 (BMP-2). In vitro cellular and electrophysiologic experiments showed that these differentiated MSCs expressing cardiomyocyte-specific markers were able to make a coupling with cardiomyocytes but not to selfbeat. The pathophysiologic significance of in vitro results was confirmed using the rat AMI model. The protein amount of TGF-beta(1) and BMP-2 in myocardium of AMI was significantly higher than that in normal myocardium. When MSCs were transplanted to the heart and analyzed 8 weeks later, they expressed cardiomyocyte-specific markers, leading to improved cardiac function. These in vitro and in vivo results suggest that infarct-related biological and physical factors in AMI induce commitment of MSCs to cardiomyocyte like cells through TGF-beta/BMP-2 pathways. PMID- 18403757 TI - Efficient serum-free derivation of oligodendrocyte precursors from neural stem cell-enriched cultures. AB - Oligodendrocytes derived in the laboratory from stem cells have been proposed as a treatment for acute and chronic injury to the central nervous system. Platelet derived growth factor (PDGF) receptor alpha (PDGFRalpha) signaling is known to regulate oligodendrocyte precursor cell numbers both during development and adulthood. Here, we analyze the effects of PDGFRalpha signaling on central nervous system (CNS) stem cell-enriched cultures. We find that AC133 selection for CNS progenitors acutely isolated from the fetal cortex enriches for PDGF-AA responsive cells. PDGF-AA treatment of fibroblast growth factor 2-expanded CNS stem cell-enriched cultures increases nestin(+) cell number, viability, proliferation, and glycolytic rate. We show that a brief exposure to PDGF-AA rapidly and efficiently permits the derivation of O4(+) oligodendrocyte-lineage cells from CNS stem cell-enriched cultures. The derivation of oligodendrocyte lineage cells demonstrated here may support the effective use of stem cells in understanding fate choice mechanisms and the development of new therapies targeting this cell type. PMID- 18403758 TI - Shared genetic causes of cardiac hypertrophy in children and adults. AB - BACKGROUND: The childhood onset of idiopathic cardiac hypertrophy that occurs without a family history of cardiomyopathy can portend a poor prognosis. Despite morphologic similarities to genetic cardiomyopathies of adulthood, the contribution of genetics to childhood-onset hypertrophy is unknown. METHODS: We assessed the family and medical histories of 84 children (63 boys and 21 girls) with idiopathic cardiac hypertrophy diagnosed before 15 years of age (mean [+/ SD] age, 6.99+/-6.12 years). We sequenced eight genes: MYH7, MYBPC3, TNNT2, TNNI3, TPM1, MYL3, MYL2, and ACTC. These genes encode sarcomere proteins that, when mutated, cause adult-onset cardiomyopathies. We also sequenced PRKAG2 and LAMP2, which encode metabolic proteins; mutations in these genes can cause early onset ventricular hypertrophy. RESULTS: We identified mutations in 25 of 51 affected children without family histories of cardiomyopathy and in 21 of 33 affected children with familial cardiomyopathy. Among 11 of the 25 children with presumed sporadic disease, 4 carried new mutations and 7 inherited the mutations. Mutations occurred predominantly (in >75% of the children) in MYH7 and MYBPC3; significantly more MYBPC3 missense mutations were detected than occur in adult onset cardiomyopathy (P<0.005). Neither hypertrophic severity nor contractile function correlated with familial or genetic status. Cardiac transplantation and sudden death were more prevalent among mutation-positive than among mutation negative children; implantable cardioverter-defibrillators were more frequent (P=0.007) in children with family histories that were positive for the mutation. CONCLUSIONS: Genetic causes account for about half of presumed sporadic cases and nearly two thirds of familial cases of childhood-onset hypertrophy. Childhood onset hypertrophy should prompt genetic analyses and family evaluations. PMID- 18403760 TI - Asthma and chitinases. PMID- 18403759 TI - Effect of variation in CHI3L1 on serum YKL-40 level, risk of asthma, and lung function. AB - BACKGROUND: The chitinase-like protein YKL-40 is involved in inflammation and tissue remodeling. We recently showed that serum YKL-40 levels were elevated in patients with asthma and were correlated with severity, thickening of the subepithelial basement membrane, and pulmonary function. We hypothesized that single-nucleotide polymorphisms (SNPs) that affect YKL-40 levels also influence asthma status and lung function. METHODS: We carried out a genomewide association study of serum YKL-40 levels in a founder population of European descent, the Hutterites, and then tested for an association between an implicated SNP and asthma and lung function. One associated variant was genotyped in a birth cohort at high risk for asthma, in which YKL-40 levels were measured from birth through 5 years of age, and in two populations of unrelated case patients of European descent with asthma and controls. RESULTS: A promoter SNP (-131C-->G) in CHI3L1, the chitinase 3-like 1 gene encoding YKL-40, was associated with elevated serum YKL-40 levels (P=1.1 x 10(-13)), asthma (P=0.047), bronchial hyperresponsiveness (P=0.002), and measures of pulmonary function (P=0.046 to 0.002) in the Hutterites. The same SNP could be used to predict the presence of asthma in the two case-control populations (combined P=1.2 x 10(-5)) and serum YKL-40 levels at birth (in cord-blood specimens) through 5 years of age in the birth cohort (P=8.9 x 10(-3) to 2.5 x 10(-4)). CONCLUSIONS: CHI3L1 is a susceptibility gene for asthma, bronchial hyperresponsiveness, and reduced lung function, and elevated circulating YKL-40 levels are a biomarker for asthma and decline in lung function. PMID- 18403761 TI - Images in clinical medicine. Cerebral embolism of probable aortic origin. PMID- 18403762 TI - California dreamin'--state health care reform and the prospect for national change. PMID- 18403763 TI - Universal coverage one head at a time--the risks and benefits of individual health insurance mandates. PMID- 18403764 TI - Challenges to HIV prevention--seeking effective measures in the absence of a vaccine. PMID- 18403765 TI - Long-term results of carotid stenting versus endarterectomy in high-risk patients. AB - BACKGROUND: We previously reported that, in a randomized trial, carotid stenting with the use of an emboli-protection device is not inferior to carotid endarterectomy for the treatment of carotid artery disease at 30 days and at 1 year. We now report the 3-year results. METHODS: The trial evaluated carotid artery stenting with the use of an emboli-protection device as compared with endarterectomy in 334 patients at increased risk for complications from endarterectomy who had either a symptomatic carotid artery stenosis of at least 50% of the luminal diameter or an asymptomatic stenosis of at least 80%. The prespecified major secondary end point at 3 years was a composite of death, stroke, or myocardial infarction within 30 days after the procedure or death or ipsilateral stroke between 31 days and 1080 days (3 years). RESULTS: At 3 years, data were available for 260 patients (77.8%), including 85.6% of patients in the stenting group and 70.1% of those in the endarterectomy group. The prespecified major secondary end point occurred in 41 patients in the stenting group (cumulative incidence, 24.6%; Kaplan-Meier estimate, 26.2%) and 45 patients in the endarterectomy group (cumulative incidence, 26.9%; Kaplan-Meier estimate, 30.3%) (absolute difference in cumulative incidence for the stenting group, 2.3%; 95% confidence interval, -11.8 to 7.0). There were 15 strokes in each of the two groups, of which 11 in the stenting group and 9 in the endarterectomy group were ipsilateral. CONCLUSIONS: In our trial of patients with severe carotid artery stenosis and increased surgical risk, no significant difference could be shown in long-term outcomes between patients who underwent carotid artery stenting with an emboli-protection device and those who underwent endarterectomy. (ClinicalTrials.gov number, NCT00231270 [ClinicalTrials.gov].). PMID- 18403766 TI - Recent resurgence of mumps in the United States. AB - BACKGROUND: The widespread use of a second dose of mumps vaccine among U.S. schoolchildren beginning in 1990 was followed by historically low reports of mumps cases. A 2010 elimination goal was established, but in 2006 the largest mumps outbreak in two decades occurred in the United States. METHODS: We examined national data on mumps cases reported during 2006, detailed case data from the most highly affected states, and vaccination-coverage data from three nationwide surveys. RESULTS: A total of 6584 cases of mumps were reported in 2006, with 76% occurring between March and May. There were 85 hospitalizations, but no deaths were reported; 85% of patients lived in eight contiguous midwestern states. The national incidence of mumps was 2.2 per 100,000, with the highest incidence among persons 18 to 24 years of age (an incidence 3.7 times that of all other age groups combined). In a subgroup analysis, 83% of these patients reported current college attendance. Among patients in eight highly affected states with known vaccination status, 63% overall and 84% between the ages of 18 and 24 years had received two doses of mumps vaccine. For the 12 years preceding the outbreak, national coverage of one-dose mumps vaccination among preschoolers was 89% or more nationwide and 86% or more in highly affected states. In 2006, the national two-dose coverage among adolescents was 87%, the highest in U.S. history. CONCLUSIONS: Despite a high coverage rate with two doses of mumps-containing vaccine, a large mumps outbreak occurred, characterized by two-dose vaccine failure, particularly among midwestern college-age adults who probably received the second dose as schoolchildren. A more effective mumps vaccine or changes in vaccine policy may be needed to avert future outbreaks and achieve the elimination of mumps. PMID- 18403768 TI - Images in clinical medicine. Pseudoaneurysm of the right internal thoracic artery. PMID- 18403767 TI - The challenge of HIV-1 subtype diversity. PMID- 18403769 TI - Case records of the Massachusetts General Hospital. Case 11-2008. A 45-year-old man with changes in mental status after liver transplantation. PMID- 18403770 TI - Clinical decisions. Management of carotid stenosis. PMID- 18403771 TI - Perspectives on medical outsourcing and telemedicine--rough edges in a flat world? PMID- 18403772 TI - Activated protein C and diabetic nephropathy. PMID- 18403773 TI - Time to defibrillation after in-hospital cardiac arrest. PMID- 18403774 TI - Breast-cancer stromal cells with TP53 mutations. PMID- 18403775 TI - Paclitaxel plus bevacizumab for metastatic breast cancer. PMID- 18403776 TI - Platelet activation and atherothrombosis. PMID- 18403777 TI - Expansion of the health workforce and the HIV epidemic. PMID- 18403778 TI - Lead poisoning due to adulterated marijuana. PMID- 18403779 TI - DNA vector augments inflammation in epithelial cells via EGFR-dependent regulation of TLR4 and TLR2. AB - Gene delivery applications to treat lung diseases are, in some instances, suboptimal due to deleterious host inflammatory reactions. Current DNA plasmids (pDNA) exert toxicity in part via unmethylated CpG motifs that stimulate Toll like receptor (TLR)9-expressing leukocytes; however, the airway epithelial response has not been well defined. Bronchial epithelial cells (BEAS-2B) were exposed to pDNA complexes and inflammatory mediators were measured. As patients with inflammatory lung disease are susceptible to infectious exacerbations, we also evaluated the reciprocal inflammatory response to pDNA and bacterial components lipopolysaccharide (LPS) and lipoteichoic acid (LTA), recognized by TLR4 and TLR2, respectively. Cells primed with pDNA synergistically expressed IL 8 mRNA and protein in response to LPS and LTA (3- to 5-fold). A similar induction was also observed for IL-1beta, IL-6, colony-stimulating factor (CSF)-1, and granulocyte macrophage-CSF. Their synergistic elevation was associated with an increase in TLR4 and TLR2 levels. Methylation of pDNA only partially reduced (25 30%) IL-8 release; hence, signaling occurs via CpG/TLR9-dependent and independent modules. As epidermal growth factor receptor (EGFR) signaling has been implicated in bronchial IL-8 expression, we assessed whether pDNA priming events were coordinated via EGFR. AG1478 (EGFR inhibitor) restored normal TLR4/2 levels and also suppressed synergistic release of IL-8. The extracellular signal regulated kinase (Erk) mitogen-activated protein kinase inhibitor also blocked IL 8 release, implicating Erk as a key mediator of EGFR signaling. Our findings identify a novel EGFR-dependent mechanism for regulating TLR, and show that targeted disruption of EGFR signaling ameliorates the airway epithelial inflammatory response to pDNA. Targeting the EGFR system may improve the efficiency, tolerability, and safety of gene therapy strategies. PMID- 18403780 TI - Functional expression of GABAB receptors in airway epithelium. AB - Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian central nervous system and exerts its actions via both ionotropic (GABA(A)) and metabotropic (GABA(B)) receptors. The GABA(B) receptor is a dimer composed of R1 and R2 components and classically couples to the heterotrimeric G(i) protein. In addition to their location on neurons, GABA and functional GABA(B) receptors have been detected in peripheral tissue such as airway smooth muscle. We questioned whether airway epithelium expresses receptors that could respond to GABA. We detected the mRNA encoding multiple-splice variants of the GABA(B)R1 and GABA(B)R2 in total RNA isolated from native human and guinea pig airway epithelium and human airway epithelial cell lines (BEAS-2B and H441). Immunoblots identified the GABA(B)R1 and GABA(B)R2 proteins in both guinea pig airway epithelium and BEAS-2B cells. The expression of GABA(B)R1 protein was immunohistochemically localized to basal mucin-secreting and ciliated columnar epithelial cells in guinea pig trachea. Baclofen inhibited adenylyl cyclase activity, induced ERK phosphorylation and cross-regulated phospholipase C, leading to increased inositol phosphates in BEAS-2B cells in a pertussis toxin sensitive manner, implicating G(i) protein coupling. Thus, these receptors couple to G(i) and cross-regulate the phospholipase C/inositol phosphate pathway. The second messengers of these pathways, cyclic AMP and calcium, play pivotal roles in airway epithelial cell primary functions of mucus clearance. Furthermore, the enzyme that synthesizes GABA, glutamic acid decarboxylase (GAD65/67), was also localized to airway epithelium. GABA may modulate an uncharacterized signaling cascade via GABA(B) receptors coupled to G(i) protein in airway epithelium. PMID- 18403781 TI - Identification of transforming growth factor beta1-driven genetic programs of acute lung fibrosis. AB - Lung fibrosis is characterized by excessive accumulation of extracellular matrix components leading to progressive airflow limitation. Distinct profibrotic pathways converge on the activation of transforming growth factor-beta (TGF beta), a central growth factor implicated in most fibroproliferative diseases. Recently, enforced expression of bioactive human TGF-beta1 (hTGF-beta1) in lungs of transgenic mice was shown to recapitulate several key pathophysiologies observed in fibrotic disorders of the lung, including cellular inflammation, tissue fibrosis, and myofibroblast hyperplasia. Inducible expression of hTGF beta1 in this system provided a unique opportunity to characterize TGF-beta driven mechanisms that precede and/or follow the onset of inflammation and fibrosis. Using gene expression profiling in lungs, we demonstrate temporal activation of key genetic programs regulating cell movement and invasiveness, inflammation, organ remodeling, and fibrosis. Consistent with our gene expression data, multiple soluble mediators associated with inflammation and tissue remodeling were markedly elevated in the bronchoalveolar lavage fluid of mice expressing hTGF-beta1. We observe significant TGF-beta1-driven infiltration of F4/80+ mononuclear cells producing bioactive arginase, a marker of alternatively activated macrophages. Finally, we identified a common "fibrosis" gene signature when comparing our findings with published data derived from preclinical and clinical studies. PMID- 18403783 TI - Does enrollment in a CDHP stimulate cost-effective utilization? AB - Consumer-driven health plans (CDHPs) are built on the assumption that with increased cost sharing consumers will select cost-effective evidence-based care. In this study, the authors explore whether patterns of utilization change after enrollment in a CDHP and whether the pattern reflects a shift toward evidence based care. The study population is comprised of 18,025 employees and their adult dependents. The analysis uses a schema for categorizing claims data into high priority (evidence-based care) and low-priority (limited or no evidence-based care) utilization. The findings indicate that enrollment in CDHPs resulted in a reduction of office visits in the 1st year of enrollment. These reductions in care appear to be indiscriminant, with patients cutting back in both high-and low priority visits. The reductions in high- and low-priority visits were greater for employees with lower education and income. PMID- 18403784 TI - Management of patients with high sperm DNA damage. PMID- 18403782 TI - Insights from the complete genome sequence of Mycobacterium marinum on the evolution of Mycobacterium tuberculosis. AB - Mycobacterium marinum, a ubiquitous pathogen of fish and amphibia, is a near relative of Mycobacterium tuberculosis, the etiologic agent of tuberculosis in humans. The genome of the M strain of M. marinum comprises a 6,636,827-bp circular chromosome with 5424 CDS, 10 prophages, and a 23-kb mercury-resistance plasmid. Prominent features are the very large number of genes (57) encoding polyketide synthases (PKSs) and nonribosomal peptide synthases (NRPSs) and the most extensive repertoire yet reported of the mycobacteria-restricted PE and PPE proteins, and related-ESX secretion systems. Some of the NRPS genes comprise a novel family and seem to have been acquired horizontally. M. marinum is used widely as a model organism to study M. tuberculosis pathogenesis, and genome comparisons confirmed the close genetic relationship between these two species, as they share 3000 orthologs with an average amino acid identity of 85%. Comparisons with the more distantly related Mycobacterium avium subspecies paratuberculosis and Mycobacterium smegmatis reveal how an ancestral generalist mycobacterium evolved into M. tuberculosis and M. marinum. M. tuberculosis has undergone genome downsizing and extensive lateral gene transfer to become a specialized pathogen of humans and other primates without retaining an environmental niche. M. marinum has maintained a large genome so as to retain the capacity for environmental survival while becoming a broad host range pathogen that produces disease strikingly similar to M. tuberculosis. The work described herein provides a foundation for using M. marinum to better understand the determinants of pathogenesis of tuberculosis. PMID- 18403785 TI - Statement on publishing clinical trials in Indian biomedical journals. PMID- 18403786 TI - Challenges in identifying genetic risk factors for common multifactorial disorders. PMID- 18403787 TI - Chemokines in human immunodeficiency virus (HIV) infection. PMID- 18403788 TI - Evaluation of nuclear DNA damage in human spermatozoa in men opting for assisted reproduction. AB - Diagnosis of sperm DNA integrity of semen sample is important for consistently high reproductive efficiency. The conventional parameters of semen analysis take into account morphology, motility, and concentration of spermatozoa in the sample, which are insufficient for evaluation of reproductive potential. Current studies have implicated abnormal organization of genomic material in sperms as a probable cause in 20 per cent cases of male infertility. This is especially important in the era of assisted reproduction technique (ART) when a majority of infertile couples opt for assisted reproduction and in where cases DNA integrity is a better diagnostic and prognostic marker as compared to routine semen analysis. This article reviews and discusses some of the current techniques employed for evaluating chromatin structure or DNA damage in spermatozoa. These different techniques include single cell gel electrophoresis (COMET assay), Terminal tranferase dUTP Nick End Labelling (TUNEL), sperm chromatin structure assay (SCSA), In situ nick translation (ISNT) and acridine orange test. These techniques are independent measure of sperm quality and assist in semen quality assessment by detecting defects in DNA integrity or chromatin structure. The discussed techniques vary in their level of accuracy, cost input, sophistication of analysis and their application depends upon the sensitivity required for analysis. The article also briefly outlines the DNA packaging and the causes of DNA damage in spermatozoa. During chromatin packing 85 per cent of the histones are replaced by protamine while the residual histones act as marker of genes which are expressed in early embryonic development. Among the different aetiological factors observed to be responsible for DNA damage in human spermatozoa increased reactive oxygen species (ROS), oxidative stress is highly correlated with greater DNA fragmentation index (DFI). Oxidative stress leads to single and double strand breaks in sperm DNA. Apoptosis and abnormal chromatin packing also contribute to DNA damage. The significance of chromatin structure studies is more stressed owing to the greater awareness to transmission of genetic diseases because of higher incidence of gene imprinting defects, increased cancer frequency and other congenital and non-congenital defects in children conceived through assisted reproduction techniques. PMID- 18403789 TI - Screening for Y-chromosome microdeletions in infertile Indian males: utility of simplified multiplex PCR. AB - BACKGROUND & OBJECTIVE: Analysis of the microdeletions in the azoospermia factor (AZF) region of Y chromosome by PCR is an important screening tool in the work-up of infertile males opting for assisted reproductive techniques. In the present study, the Y chromosome microdeletions were analyzed by PCR using primers corresponding to 16 sequence tagged sites (STS) and three genes of the AZF region in infertile Indian men. Feasibility of developing a simplified multiplex PCR for screening of the Y chromosome microdeletions has been explored. METHODS: A total of 271 male subjects were analyzed, of which, 170 were infertile patients (51 oligospermic and 119 azoospermic) and 101 were fertile controls. Subjects showing normal karyotype only were included in the study. The semen analysis was done and plasma follicle stimulating hormone (FSH) concentrations were determined by radioimmunoassay. Testicular histopathology was analyzed by fine needle aspiration cytology (FNAC). RESULTS: Y chromosome microdeletions were observed in nine out of 170 (5.29%) infertile males all of whom were azoospermic. Of the nine subjects, two had deletions in AZFa, one in AZFb, three in AZFc and three in AZFb+c regions. No deletions were observed in the infertile severe oligospermic men (< 5 million sperm/ml semen) and fertile controls. No difference in the FSH concentrations of infertile patients with and without deletions (18.36 and 18.10 mIU/ml respectively) was observed. A clear relationship between Y chromosome microdeletions and testicular phenotypes could not be established. Two multiplex PCRs were designed using 7 STSs markers, which could detect Y chromosome microdeletions in infertile male subjects as efficiently as PCR based on larger number of PCR reactions. INTERPRETATION & CONCLUSION: The multiplex PCRs described in the present study may be a suitable, cost-effective and less time consuming method for screening the Y chromosome deletions in infertile males in routine clinical diagnosis and counselling prior to assisted reproduction. PMID- 18403790 TI - Chemokine profile among human immunodeficiency virus-1 (HIV-1) infected individuals from southern India. AB - BACKGROUND & OBJECTIVE: Individuals infected with HIV-1 have higher levels of chemokine producing cells compared to uninfected individuals. It is important to know the changes in chemokine levels associated with rate of progression of disease. There is a paucity of information on the plasma chemokines in HIV-1 infected individuals from India. We therefore carried out this study to estimate the levels of three chemokines namely macrophage inflammatory protein alpha (MIP1alpha), MIP1beta and RANTES, in relation to disease status in HIV-1 infected individuals and compared with uninfected individuals. METHODS: RANTES and MIP1alpha were estimated using ELISA in 114 HIV-1 infected and 30 controls, whereas MIP1beta was estimated in 101 HIV infected individuals only and 30 controls. The values were compared to the T cell subsets, HIV-1 viral loads and plasma cytokines (interferon gamma and interleukin-10). RESULTS: Compared to controls the mean MIP1alpha and RANTES level among the HIV-1 infected individuals was higher while MIP1beta level was lower in HIV infected individuals except CDC C groups. There was a significant positive correlation for MIP1a with HIV-1 viral load and IFNgamma, for MIP1alpha with viral load and IL10. There was a significant negative correlation between MIP1alpha with CD4 count and CD4: CD8 ratio and MIP1beta with CD4 count and CD8 count. There was a negativecorrelation between RANTES values and CD8 per cent. INTERPRETATION & CONCLUSION: In conclusion, our study showed a significantly higher level of beta chemokines in south Indian HIV-1 infected individuals compared to controls. These beta chemokines may have the inhibitory effect on HIV-1 only during the initial period and with the progression of disease this inhibitory effect wanes as shown by the positive correlation of beta chemokines with HIV-1 viral load. PMID- 18403791 TI - Multicenter evaluation of individual donor nucleic acid testing (NAT) for simultaneous detection of human immunodeficiency virus -1 & hepatitis B & C viruses in Indian blood donors. AB - BACKGROUND & OBJECTIVE: India has a high prevalence of HIV-1, hapatitis C and B virus (HCV and HBV) in the blood donors but has yet to implement nucleic acid testing (NAT) in blood screening. We undertook a multicentre evaluation of blood donor testing by NAT for simultaneous detection of HIV-1, HBV and HCV in a single tube and also to determine the feasibility of NAT implementation in India's low volume setting. METHODS: A total of 12,224 unlinked samples along with their serological results were obtained from representative eight blood banks in India and were individually manually tested by the Procleix Ultrio Assay (Chiron Corp. Emeryville, CA) for simultaneous detection of HIV-1, HCV, and HBV. RESULTS: Of the 12,224 samples tested, 209 (1.71%) were seroreactive. One hundred thirty three samples (1.09%) were reactive by Ultrio assay, 84 samples were seroreactive but NAT non reactive. There were eight NAT yield cases: 1 HIV, 1 HIV-HCV co infection, and 6 HBV. INTERPRETATION & CONCLUSION: Our observed NAT yield for all three viruses was 1 in 1528 (0.065%). We estimate NAT could interdict 3272 infectious donations a year among our approximate 5 million annual donations. PMID- 18403792 TI - Commitment among state health officials & its implications for health sector reform: lessons from Gujarat. AB - BACKGROUND & OBJECTIVE: Commitment, competencies and skills of people working in the health sector can significantly impact the performance and its reform process. In this study we attempted to analyse the commitment of state health officials and its implications for human resource practices in Gujarat. METHODS: A self-administered questionnaire was used to measure commitment and its relationship with human resource (HR) variables. Employee's organizational commitment (OC) and professional commitment (PC) were measured using OC and PC scale. Fifty five medical officers from Gujarat participated in the study. RESULTS: Professional commitment of doctors (3.21 to 4.01) was found to be higher than their commitment to the organization (3.01 to 3.61). Doctors did not perceive greater fairness in the system on promotion (on the scale of 5, score: 2.55) and were of the view that the system still followed seniority based promotion (score: 3.42). Medical officers were upset about low autonomy in the department with regard to reward and recognition, accounting procedure, prioritization and synchronization of health programme and other administrative activities. INTERPRETATION & CONCLUSION: Our study provided some support for positive effects of progressive HR practices on OC, specifically on affective and normative OC. Following initiatives were identified to foster a development climate among the health officials: providing opportunities for training, professional competency development, developing healthy relationship between superiors and subordinates, providing useful performance feedback, and recognising and rewarding performance. For reform process in the health sector to succeed, there is a need to promote high involvement of medical officers. There is a need to invest in developing leadership quality, supervision skills and developing autonomy in its public health institutions. PMID- 18403793 TI - Plasma homocysteine levels & 677C-->T methylenetetrahydrofolate reductase gene polymorphism in patients with coronary artery disease of different severity. AB - BACKGROUND & OBJECTIVE: Numerous studies have identified hyperhomocysteinemia as an independent risk factor for coronary artery disease (CAD). Furthermore, influences of polymorphysim of methylenetetrahydrofolate reductase (MTHFR) on homocysteine levels are documented. However, the relationship between severity of CAD and polymorphism of MTHFR has not been systematically evaluated. The present study was undertaken to evaluate this relationship in patients undergoing coronary artery bypass surgery. METHODS: Serum homocysteine and MTHFR polymorphism in relation to severity of CAD was examined in 113 male patients, who all underwent coronary artery bypass surgery. The prevalences of 677 C-->T transition of the MTHFR gene were determined in these patients. Two groups were compared according to GENSINI coronary score : mild atherosclerosis (CAD stenosis < 30) and severe atherosclerosis (CAD stenosis > 30). RESULTS: Patients with CAD showed a significantly higher serum concentration of homocysteine than control subjects (P < 0.01). The serum homocysteine level was significantly higher in patients with increased scores than in patients with mild CAD (Gensini score < 30) both with and without the MTHFR polymorphism. INTERPRETATION & CONCLUSION: The findings of our study showed that hyperhomocysteinemia was significantly related to the severity of CAD independent on MTHFR polymorphism. PMID- 18403794 TI - Diagnosis of common mental disorders by using PRIME-MD Patient Health Questionnaire. AB - BACKGROUND & OBJECTIVE: Primary care physicians in their clinical settings usually fail to diagnose common mental disorders (CMDs). Primary Care Evaluation of Mental Disorders (PRIME-MD) Patient Health Questionnaire (PHQ) has been shown to facilitate diagnosis of most CMDs seen in primary health care. Its utility in Indian setting has not been evaluated. We carried out this study in Indian setting to evaluate the extent of psychiatric morbidity as measured by PRIME-MD PHQ. METHODS: A total of 500 consenting patients attending Medical outpatient department were recruited. All subjects were first assessed by the physician for presence of any physical illness and psychiatric disorder and their socio demographic data were collected. Subjects were asked to fill the PRIMEMD PHQ. Illiterate subjects were assisted by the research worker in filling up the questionnaire. RESULTS: At initial evaluation physicians opined that 30.4 per cent of the subjects had psychological disorder, the most common diagnostic category was of anxiety disorders (15.8%), followed by depression (6%) and somatoform disorders (5.6%). On PHQ 42 per cent subjects had at least one psychiatric diagnosis. The most common psychiatric diagnosis was panic disorder (18.4%), followed by other anxiety disorders (16.6%). On comparing the diagnosis of physicians and PHQ, out of 152 cases (30.4%) suspected to have any psychological disorder by the physicians in the initial evaluation, 105 were found to have PHQ diagnosis. Of the 348 cases in which physicians did not suspect any psychiatric diagnosis, 243 were also negative in PHQ screening. There was a significant correlation between physicians and PHQ diagnosis. INTERPRETATION & CONCLUSION: There is a high psychiatric morbidity in the general medical practice and in many cases psychiatric morbidity is either missed or is misdiagnosed and by the physicians. Our results showed that PHQ could be a valuable screening instrument for psychiatric morbidity in primary care and general medical practice. PMID- 18403795 TI - Effect of single oral dose of sodium rabeprazole on the intragastric pH & volume in patients undergoing elective surgery. AB - BACKGROUND & OBJECTIVE: While evaluating the effectiveness of drugs used for the prophylaxis of acid aspiration of gastric contents, the impact of duodeno-gastric reflux on gastric contents has not been studied earlier. This study was carried out to evaluate the effect of preanaesthetic oral administration of sodium rabeprazole on pH and volume of gastric contents in adult patients undergoing elective surgery by excluding cases contaminated with duodeno-gastric refluxate. METHODS: The patients in group C (control) in the triple blind placebo controlled trial received placebo while group S sodium rabeprazole 20 mg orally at 2100 h, a night before elective surgery. Next day, gastric contents were aspirated with a large bore, multi-orifices gastric tube passed through an endotracheal tube placed blindly in oesophagus after tracheal intubation and analyzed for the presence of bile salts, pH and volume. The pH and volume of gastric contents were the primary and duodeno-gastric reflux secondary outcome measures of the study. RESULTS: The pH and volume of group S-2 were 3.97+/-1.78 and 9.48+/-8.39 ml respectively compared with 1.90+/-0.47 and 19.60+/-18.56 ml of group C-2. Sodium rabeprazole, after excluding contaminated cases with duodeno-gastric refluxate, significantly increased the pH (P<0.001), decreased the volume of gastric contents (P<0.005) and the proportion of the patients (30.76 vs 2.63%) considered at risk compared with placebo (P<0.001) according to the criteria defined (pH < 2.5 and volume > 25 ml). Thirty nine samples (33.33%) out of 117 were contaminated with duodenal contents. Duodenogastric reflux significantly (P<0.001) affected pH and volume of gastric in both groups C-1 vs C-2 and S-1 vs S-2. INTERPRETATION & CONCLUSION: Sodium rabeprazole 20 mg given orally a night before surgery provided adequate prophylaxis for acid aspiration syndrome at the time of induction of anaesthesia and duodeno-gastric reflux significantly affected both the pH and volume of gastric contents. PMID- 18403796 TI - Characterization of Streptococcus pneumoniae ophthalmic, systemic & commensal isolates by pulsed-field gel electrophoresis & ribotyping. AB - BACKGROUND & OBJECTIVE: Streptococcus pneumoniae is common in ocular and systemic infections and is a part of normal nasopharyngeal flora. Very few studies regarding genetic analysis of S. pneumoniae isolates causing eye infections are available. This study was undertaken to do pulse field gel electrophoresis (PFGE) analysis and ribotyping of S. pneumoniae isolates obtained from eye infections, systemic infections and nasopharyngeal flora. METHODS: Sixty one well characterized S. pneumoniae isolates (38 from ophthalmic infections, 9 from systemic infections and 14 commensals) were characterized using PFGE of the whole genome after SmaI, restriction enzyme digestion and conventional ribotyping using Escherichia coli rRNA operon as the probe. Phylogenetic tree was drawn using unweighted pair group method analysis (UPGMA). RESULTS: The 38 S. pneumoniae isolates from eye infections belonging to 15 serotypes were placed in to 11 PFGE types and 15 ribotypes. The 9 systemic isolates (7 seotypes) were distributed in 7 PFGE types and 6 ribotypes. The 14 commensal isolates were placed in 11 serotypes, 5 PFGE types and 6 ribotypes. Most of the PFGE types and ribotypes consisting of ocular isolates also contained systemic and commensal isolates. INTERPRETATION & CONCLUSION: Considerable genetic similarity was observed between the isolates from ocular and systemic infections and those colonized in nasopharynx. PFGE analysis could differentiate majority of the isolates according to site of infections. There was a considerable DNA polymorphism within the studied bacterial population. PMID- 18403797 TI - Identification of Anopheles fauna in a hyperendemic falciparum area of Orissa State, India. AB - BACKGROUND & OBJECTIVE: Keonjhar district of Orissa State has been hyperendemic for falciparum malaria since many years with alarming deaths due to cerebral malaria. Therefore an entomological investigation to know more about the relative prevalence of Anopheles species was done. METHODS: Daytime indoor resting and outdoor resting, light trap and double bed net collections were made. Surveys were also made to collect Anopheles immature in streams and paddy fields. The Anopheles mosquitoes obtained by different catching methods were identified and the known vector species were subjected to gut and salivary gland dissection for vector incrimination. The infected specimens of An. fluviatilis and An. minimus were subjected to polymerase chain reaction assay for identification of sibling species. RESULTS: Of the anophelines collected, the most abundant was An. splendidus (18.2%) and An. fluviatilis (17.7%), followed by An. maculatus (14.0%) and An. minimus (9.0%). The sporozoite rate of An. fluviatilis and An. minimus was 0.9 and 1.4 respectively. The infected specimens have been identified as sibling species S of the An. fluviatilis complex and A of the An. minimus complex. INTERPRETATION & CONCLUSION: An. fluviatilis and An. minimus are the major two species in the transmission of malaria in Keonjhar district in Orissa. PMID- 18403798 TI - Antimicrobial activities of four Tunisian Chrysanthemum species. AB - BACKGROUND & OBJECTIVE: Tunisian Chrysanthemum species are known to have medicinal activity and some of the species are used in traditional medicine. We have earlier shown the use of C. trifurcatum flowerheads in Tunisian traditional medicine to treat constipation. In the present study we investigated the anti microbiol activity of four Tunision Chrysanthemum species. METHODS: Different parts (flowers, leaves, stems, roots, leaves and flowers and leaves and stems) of four Tunisian Chrysanthemum species, were extracted with solvents of increasing polarity to obtain aqueous and organic extracts. These extracts were tested in vitro for their antimicrobial activity against 14 bacteria and four yeasts, using agar diffusion and microdilution methods. Activity was evaluated by measuring the zones of inhibition against the tested organisms and minimum inhibitory concentrations (MIC) were determined from the lowest concentrations of extracts to inhibit the growth of microorganisms. Cytotoxity and antiviral activities against Herpes simplex virus type 1 (HSV-1), were evaluated using the neutral red incorporation method. RESULTS: Extracts of the 4 Chrysanthemum species showed some degree of activity against one or more of the microbial strains with MIC ranging from 0.625 to 1.25 mg/ml. Most of the extracts were well tolerated by Vero cells with CC(50) > 500 microg/ml. The petroleum ether extract of C. trifurcatum stems and leaves protected infected cells with EC(50) of 100 microg/ml. INTERPRETATION & CONCLUSION: Our findings showed that some Chrysanthemum extracts exhibited antimicrobial and/or anti-HSV-1 activities. Further studies aimed to the isolation and identification of active substances from the extracts which exhibited interest activities, need to be done. PMID- 18403799 TI - Potantial utility of disulfiram against leishmaniasis. PMID- 18403800 TI - Evaluation of CIVA agar for rapid detection of extended spectrum beta- lactamases (ESBL) among isolates of Enterobacteriaceae. PMID- 18403801 TI - Impact Factor of the IJMR: what more we really need? PMID- 18403803 TI - Autism, the big unkown. AB - Autism is a common disorder of childhood. Yet, it often remains unrecognized and undiagnosed until or after late preschool age because appropriate tools for routine developmental screening and screening specifically for autism have not been available. Paediatricians have an important role in early recognition and evaluation of autism spectrum disorders because they usually are the first point of contact for parents. It is important that paediatricians are able to recognize the signs and symptoms of autism spectrum disorders and have a strategy for assessing them systematically. But paediatricians have also a role in chronic management of these disorders. The objective of this paper is to show a general view of the autism spectrum disorders (ASD) state of knowledge nowadays as well to stress the need of early detection and treatment of these disorders in order to improve better evolution and prognosis. PMID- 18403804 TI - [Is there are needs treating the adolescents with syndrome of vegetative dysfunction?]. AB - Article presents the pathogenesis aspects and clinical data of adolescents with syndrome of vegetative dysfunction. The authors observe the structure of different clinical forms, reflect on conditions and situation, when adolescents needs a stable complex of etiopathogenetic therapy to control autonomic nervous system dysfunction. PMID- 18403805 TI - Assessment of traumatic brain injury and anterior pituitary dysfunction in adolescents. AB - Data from the Italian Ministry of Health show that approximately 300-500 per 100.000 Italians are admitted to hospital each year for either TBI or subarachnoid haemorrhage with an annual mortality of 20 per 100.000; 90% of these TBI are of medium severity. Traumatic brain injury-induced hypopituitarism in adults are more common than previously thought. The paucity of clinical reports relating to adolescents with past-TBI induced hypothalamic-pituitary-dysfunction suggests that this phenomenon might be less common that that observed in adults. In the last 25 years, in our Unit a pituitary dysfunction was established during childhood and adolescence in 3 patients (one patient had a precocious puberty, one patient had a gonadal dysfunction and one patient had a partial growth hormone deficiency). In all patients the TBI was severe (unpublished data, 2008). The physiopathological basis of hypopituitarism is lacking. Nevertheless, necrotic, hypoxic, ischemic and shearing lesions are at the hypothalamus and/or the pituitary are likely important factors. The subjects at highest risk appear to be those who have suffered a moderate or severe trauma. Clinical signs of anterior hypopituitarism are often subtle and may be masked by sequalae of TBI. Therefore, post-traumatic anterior pituitary dysfunction may remain undiagnosed and, possibly, aggravate symptoms of brain injury. Moreover it may, if undiagnosed, lead to potentially fatal endocrine crisis. Therefore, adolescents with moderate-severe traumatic brain injury should be screened for such endocrine deficiencies so that replacement therapy can be initiated to optimized the rehabilitation and outcome. PMID- 18403806 TI - [Coenzyme q10 in childhood: detection methods, reference values and disease related changes in the coenzyme q10 status]. AB - The lipophilic antioxidant coenzyme Q10 is an effective inhibitor of oxidative damage. Furthermore coenzyme Q10 is involved in electron transport related to the mitochondrial respiratorial chain. Because of this double function coenzyme Q10 has become a special role in the group of antioxidants. Little is known about coenzyme Q10 in healthy and sick children. The aim of the study was to determine the role of coenzyme Q10 in the pathophysiological concept of pediatric diseases. At first a HPLC-method for the detection of coenzyme Q10 in plasma, erythrocytes and platelets was developed and age-related reference values for children were established. Based on these reference values the CoQ10 status was measured in different pediatric diseases. By this way various conditions for low coenzyme Q10 plasma values in children could be defined. Furthermore there were different in vivo models developed to define pharmacokinetic and pharmacodynamic characteristics of coenzyme Q10. The established methods and measured data might be a helpful contribution for estimating coenzyme Q10 deficiency and for planning therapeutical studies with coenzyme Q10 in childhood. PMID- 18403807 TI - Comparative efficiency of diagnostics and treatment for Helicobacter pylori infection in children. AB - The prevalence of chronic gastroenterological diseases among schoolchildren is high and the incidence of those diseases is growing up. The article is devoted to study of diagnostic, clinical features of Helicobacter pylori associated chronic diseases of upper digestive tract in children. With a help of screening questioning of school children the age prevalence of main, gastroduodenal specific complaints has been determined, their social and everyday life peculiarities have been investigated. Comparative efficacy of the main invasive and non invasive Helicobacter pylori diagnostic techniques among children has been determined. With the help of qualitative Helicobacter pylori stool antigen test the comparative efficacy of different methods of eradication therapy based on using bismuth containing drugs and the proton pump inhibitors were studied. The eradication therapy tolerability and side effects profile have been investigated. PMID- 18403808 TI - Analysis of clinical experience of using formula nutrilon for bottle feeding of the first year of life in Ukraine. AB - The article describes some current issues of bottle feeding of first year children. Human milk oligosaccharides play an important role in postnatal development of the intestinal flora and development of protective functions of child's organism. Dietary modulation of the intestinal microflora and immune response is one of the important problems in the nutritional sciences today. The present review summarizes the data of experimental research and clinical studies concerning the possible effects of probiotic mixture of galacto-oligosaccharides and fructo-oligosaccharides in Ukraine. The data demonstrate that prebiotic oligosaccharides such as studied mixture provide beneficial effects for formula fed infants. The results from several studies in Ukraine demonstrate that probiotic oligosaccharides stimulate the growth of bifidobacteria and lactobacilli, reduce the growth of pathogens, decrease faecal pH, normalize the stool consistency and modulate immune system as human milk does. PMID- 18403809 TI - [Sleep problems in childhood: basis of sleep regulation and intervention possibilities]. AB - In the article there is provided the up-to-date information about fundamentals of sleep regulation in childhood, sleep disturbances and treatment possibilities as well. PMID- 18403810 TI - Feeding difficulties in children and adolescents with chronic illness. AB - Good health demands good nutrition and in the child it is reflected in normal growth. Children who cannot or do not eat properly often become unwell and do not grow. This becomes a source of great concern and anxiety for their parents. Several chronic illnesses in children impair normal feeding; this article aims to describe the interrelationship between eating and disease in children with reference to some common conditions. The effects of childhood eating disorders on parents and families will also be considered. PMID- 18403811 TI - Adolescent medicine: from clinical practice to public health. AB - In most countries, adolescent health problems have shifted from pure medical conditions to more psychosocial burdens such as injuries and violence, substance use, unsafe sex and chronic conditions including under nutrition or obesity. This new situation requires specific actions which have to take into account the specificities of the bio psychosocial development of the adolescent. Youth friendly services offering adequate environment and policies as well as carefully trained physicians represent one answer to the health needs of adolescents. Another lies in the development of school prevention and health promotion. Finally, policies aiming at securing a safe environment represent an effective mean to improve the health of adolescents. PMID- 18403812 TI - New approach to estimate different drugs and/or other medical interventions effectiveness based on fuzzy logic principles. AB - A new approach for the evaluation of the efficacy of drugs and/or other medical interventions is proposed. It is based on the principles of fuzzy logic, particularly on fuzzy sets, fuzzy cluster analysis and fuzzy expert system. PMID- 18403813 TI - Network in pediatric rheumatology: the example of pediatric rheumatology international trials organisation. AB - The pediatric rheumatic diseases (PRD) are rare conditions associated with important sequelae on the quality of life and long term outcome. The research aimed at studying new therapeutic approaches is difficult because of logistic, methodological and ethical problems. To face these problems 2 international networks; the Pediatric Rheumatology Collaborative Study Group (PRCSG) and the Paediatric Rheumatology International Trials Organization (PRINTO) have been founded. The 2 networks have the goal to promote, facilitate and conduct high quality research for the PRD. In particular they have been able to standardize the evaluation of response to therapy in juvenile idiopathic arthritis (JIA), juvenile systemic lupus erythematosus, and juvenile dermatomyositis, to draft clinical remission criteria in JIA, and to provide cross-cultural adapted and validated quality of life instruments like the Childhood Health Assessment Questionnaire, and the Child Health Questionnaire, into 32 different languages. In this paper we reviewed how the creation of large international trial networks such as PRINTO and PRCSG, the definition of internationally recognized and standardized outcome measures and definitions of improvement, the validation of quality of life instruments, the adoption of adequate legislative measures (pediatric rule), have created the basic premises for the best future assessment of the PRD. This progress now offers children with PRD the same opportunities as adults to be treated with drugs whose safety and efficacy have been assessed through legitimate scientifically valid investigations. PMID- 18403814 TI - Spermatogenesis in young adult patients with beta-thalassaemia major long-term treated with desferrioxamine. AB - Since the introduction of hypertransfusion and intensive iron chelation therapy, patients with homozygous beta-thalassaemia major (TM) achieve adulthood. Many patients grow and develop normal hoping for marriage and to have a family. Therefore the question of fertility potential in this adult group of TM patients has become paramount. We report the semen parameters, the endocrine functions and serum zinc levels in 12 young adult TM patients. Their mean age was 24.8 years. Six patients (50%) had a normal sperm count, motility and morphology. While the remaining patients had oligospermia (sperm concentration <20 x 10(6)/ml) and/or asthenospermia (motility <40%). Basal serum gonadotrophins [LH and FSH], total and free testosterone and serum zinc did not differ significantly from those found in 13 normal adults with comparable testicular size. At the time of the study serum ferritin levels ranged from 240 to 3055 ng/ml (mean 1139 ng/ml). No correlations were found between semen parameters, serum total and free testosterone, plasma zinc, serum ferritin and seminal parameters. Nevertheless we observed that serum ferritin levels were lower (mean 543 ng/ml) in TM patients with abnormal seminal parameters (count and motility) compared to TM patients with normal seminal parameters (mean serum ferritin 1276 ng/ml; p<0.01). In conclusion, impairment of semen parameters may be a negative effect of intensive chelation therapy. Clearly, further investigations are required to evaluate if these adverse effects can be reduced or prevented, and if the existing spermatogenesis damage is reversible. PMID- 18403815 TI - [Neuroimmune regulation in children with bronchial asthma]. AB - 80 children aged from 2 to 14 years,72 boys and 8 girls, 44 with moderate and 36 with severe form of atopic bronchial asthma were investigated. Substance P, neurokinin A and vasoactive intestinal peptide (VIP) were defined in blood plasma. In comparison with the control group, children suffering from bronchial asthma showed statistically significant (p<0,001) increase of substance P and neurokinin A and decrease of VIP. Analogous changes were observed by comparison of data received from children with moderate and severe asthma. Received data indicate the participation of neuropeptides in pathogenesis of bronchial asthma in children. PMID- 18403816 TI - Simulated-patient-based programs for teaching communication with adolescents: the link between guidelines and practice. AB - Adolescents frequently tend not to share personal issues with their health care providers, thus communication with the adolescent patient and frequently also with his or her parents requires unique skills on behalf of the physician. Guidelines for obtaining information regarding adolescents' risk behaviors and other psychosocial issues that have been developed, do not provide the necessary tools for optimal communication with adolescents. Communication skills are best obtained in role-play models where either colleagues or actors simulate the patients' roles. Simulation-based medical education offers a safe and "mistake forgiving" environment that enables consideration of the trainees' needs, without the use of real patients that is associated with traditional bedside teaching. Training programs to improve physicians' communication skills with adolescents have been developed at the Israel Center for Medical Simulation (MSR). Between 2003 and 2007, 470 physicians were trained at MSR in 40 one-day courses. These courses dealt with common adolescent health issues that require unique communication skills on behalf of the clinician, utilizing the simulated-patient based programs. At each training day up to 12 physicians were exposed to 8 typical adolescent health related scenarios simulated by professional actors in rooms equipped with video facilities and one-way mirrors. Following the encounters with the simulated patients, the different scenarios were discussed in debriefing group sessions with experienced facilitators utilizing the encounters' video recording. Feedbacks from participants in the programs were excellent, emphasizing the need to include simulation-based programs in physicians' training curricula. PMID- 18403817 TI - Nutritional protein intake is associated with body mass index in young adolescents. AB - Preventing and treating obesity has become a major public health concern. Obesity results from the imbalance of energy intake and energy expenditure. Energy intake has increased, energy expenditure has decreased in recent years. In addition, the proportion of macronutrient intake has changed with a substantial decline in the percentage of energy from nutritional fat, and an increase in energy from sweetened beverages. We focus on the interaction between body mass index and the recent population wide increase in protein consumption. 7182 three-day weighing and nutritional protocols and anthropometric data of the DONALD (Dortmund Nutritional and Anthropometrical Longitudinally Designed) Study from 1028 healthy children and adolescents, aged 2 to 18 years (51.3% girls, 48.7% boys), measured between 1985 and March 2006, were obtained and re-analyzed. Mean daily energy intake (r=0.060, p>0.1), and the absolute daily intake of fat (r=0.031, p>0.1), and carbohydrate (r=0.050, p>0.1) were independent from BMI standard deviation scores (BMI-SDS). However, a significant interaction between BMI-SDS and the mean absolute daily intake of all protein (r=0.143, p<0.0001), and animal protein (r=0.151, p<0.0001) was found. When expressing macronutrient intake as percentage of daily energy intake, the fat and carbohydrate correlations remained insignificant with r= -0.040, and r= -0.037, respectively, whereas the correlation between BMI-SDS and all protein (r=0.203, p<0.0001), and animal protein (r=0.163, p<0.0001) further increased. The correlation depended on age and reached maxima in the group of 10-12 year old boys (r=0.31, p<0.0001), and girls (r=0.36, p<0.0001). Protein intake explained up to 13% of the BMI variance in young adolescents. We strongly suggest reconsidering the recommended daily allowances of nutritional protein, to abstain from the popular very high protein diets, and particularly from adding the flavouring agent monosodium glutamate. PMID- 18403818 TI - Clinical and psychological characteristics of school adaptation. AB - 70 girls and 50 boys--totally 120 practically healthy children were examined; Three age-specific groups with equal number of children were chosen. Goal of the research was to evaluate the adaptation ability and analyse the disadaptive derangements in primary school age children. Cross-sectional, one moment research was carried out according to D. Stot's "Map of Observation". Adaptive potential determination and evaluation of vegetative nervous system functional state was done. Special charts for children were filled in. 56% of pupils were practically healthy and well adapted to school obligations. In the rest of cases different degree and form of school dysadaptation derangements were revealed. The dysadaptation contingent with cognitive function was seen in 14% of children. Emotional attitude derangements to learning process was revealed in 18% of children.; and hard to cope with the derangements correction in behavior was seen in 12% of children. The study results enabled us to reveal different types of school dysadaptation in primary school age children as well as to study the main reasons for their formation on the basis of age-specific characteristics. The above-mentioned fact should be taken into consideration while elaborating complex rehabilitation actions and school disadaptation prevention. PMID- 18403820 TI - Expert diagnosis in tall stature: edits 1.1 diagnostic software efficacy. AB - EDITS (Expert Diagnosis In Tall Stature) expert system version 1.1 was evaluated by making it diagnose sets of clinical and laboratory criteria published as case reports in medical scientific literature, and analyzing the results obtained. The case reports were selected by means of the PubMed database. Processing the input data yielded a list of the top five most-probable disorders, ranked in the order of likelihood along with their probability. 21 cases of 12 diseases were processed. In 14 cases out of 21 (66.67%) the referral diagnosis was included in the top five most probable diagnoses listed by the expert system. In 64.29% of all the cases when the correct diagnosis was put on the top five list, the correct diagnosis ranked first. The efficacy of EDITS 1.1 is in line with that demonstrated by SYNDROC, POSSUM, OMIM and the Expert System for Differential Diagnosis of Hereditary Neuromuscular Diseases in Children. EDITS 1.1 diagnostic software proved to be a useful practical tool for differential diagnosis of disorders that may be manifested by the tall stature syndrome. PMID- 18403819 TI - Hematologic response to hydroxyurea therapy in children with beta-thalassemia major. AB - beta-thalassemia major is the most common monogenic hereditary blood disease in children. beta+-thalassemia major gene frequency in Georgia averages 0,019 (3,79% gene carriers). Hydroxyurea (HU) has been known to cause induction of fetal hemoglobin (HbF), but the efficacy of this treatment in beta-thalassemia patients is still unclear. This study was undertaken to evaluate the clinical and hematologic responses in patients with beta+-thalassemia to treatment with HU during 5 years in Georgia. Six children, aged 8 years to 13 years with transfusion-dependent beta+-thalassemia phenotype were enrolled in a trial to assess the response to HU therapy. Hemoglobin, reticulocyte count, HbF and ferritin were evaluated. The starting dose of HU was 5 mg/kg per day (5 days week) given orally once a day. Response to therapy was evaluated at 1, 2, and 5 years of treatment. Clinical improvement and rise in the HbF levels was observed in all patients. We report three cases of a remarkable response to treatment with HU in which the red cell transfusion was stopped after 1 year of treatment, and the patients became completely transfusion-free for more than 5 years. A moderate response was seen in two patients, who remained transfusion-dependent, but at longer intervals. There was no serious complication of treatment with HU. Long term HU therapy may correct the anemia and can eliminate or minimize the transfusional needs in children with beta+-thalassemia major in cases, when the patient's baseline HbF level is > or =15% and its increase during the treatment is up to 20%. PMID- 18403821 TI - General movements in infants with autism spectrum disorders. AB - General movements (GMs) are a distinct movement pattern carried out spontaneously without external stimulation and seen in fetuses of 9 weeks gestational age till 21 weeks postterm. GMs are helpful in the early diagnosis of an impaired central nervous system and the specific prediction of later neurological deficits. Autism spectrum disorder (ASD) is a neurodevelopmental disorder involving a life-long deficit in several aspects of the social and communicative behavior. Recently there appeared studies proving that children with ASD demonstrate disorders of motor development. To detect whether abnormalities in spontaneous motor activity can be observed already in the first months of life in infants with ASD. A retrospective study was performed by analyzing the family videos provided by parents of 20 children (male 17, female 3) later diagnosed as ASD. Home videos provided by parents of a control group of healthy children (n=20; male 10, female 10) matched for age with the ASD subjects and recorded in similar conditions were also analysed. In total 70 sequences were studied. Two independent observers, blind of the infants' outcome (ASD or normal), assessed the cases applying a global and a more detailed assessment of GMs. Hence, the age-specific GM pattern (normal or abnormal) as well as motor optimality scores were determined for each video sequence. Cohen kappa was 0.614. During the writhing movement period 70.0% sequences of infants with ASD showed poor repertoire GMs. In the control group, poor repertoire GMs were only seen in 12.5% of the sequences. In the fidgety movement period 20.8% of sequences were assessed as absent fidgety movements, 29.2% as abnormal fidgety movements. The large majority of the videos for the control cases were scored as normal (88.9%), 11.1% had no fidgety movements. According to the Mann-Whitney U test there were significant differences between the ASD and the control groups' optimality scores. The optimality scores were lower in the ASD group. The reduced optimality scores were mainly due to a lack of variable sequences, amplitude and speed of writhing GMs and an altered quality of fidgety and other spontaneous movements in the ASD group. Infants with ASD had more often poor repertoire writhing GMs as well as abnormal or absent fidgety movements than control infants. These data encourage further studies involving a larger number of family videos. PMID- 18403822 TI - Familial Mediterranean Fever in Armenian population. AB - Familial Mediterranean Fever (FMF) is an inherited, recessively transmitted inflammatory condition usually occurred in populations from Mediterranean descent (Armenian, Arab, Jewish, Greek, Turkish and Italian populations). Identification of MEFV gene mutations has been of tremendous help for early diagnosis of most cases. The frequency of FMF is different. The prevalence of heterozygous carriers of one of the mutations of MEFV gene is as high as 1 in 5 healthy individuals in Armenia. Genetic testing of this rare Mendelian disorder (MIM no 249100) is efficient for early and prenatal diagnosis of the disease, especially for atypic cases, for carrier screening and pregnancy planning since certain mutations have been shown to have significant correlation with renal amyloidosis (RA), the most severe possible manifestation of FMF. Also genetic testing is very important for colchicine therapy correction. Twelve MEFV mutations are identified in 7000 Armenian FMF patients. Investigation of MEFV mutations in FMF patients (heterozygotes, homozygotes and compound heterozygotes) in comparison with healthy individuals has revealed the most frequent mutations and genotypes, and the information was received about the heterozygous carriers and genotype phenotype correlation. In heterozygote carriers the most prevalent and severe cases are caused by the presence of a single M694V mutation. Our results could confirm that the MEFV gene analysis provides the first objective diagnostic criterion for FMF (characterisation of the two MEFV mutated alleles in more than 90% of the patients). Molecular testing is also used to screen the MEFV gene for mutations in patients with a clinical suspicion of FMF. We also demonstrated the unfavourable prognostic value of the M694V homozygous genotype, and provided the first molecular evidence for incomplete penetrance and pseudo-dominant transmission of the disease. Overall, these data, which confirm the involvement of the MEFV gene in the development of FMF, should be essential in clinical practice, leading to new ways of managment and treatment of FMF patients. PMID- 18403823 TI - A rare cause of heart failure in iron-overload thalassaemic patients-primary hypoparathyroidism. AB - Hypocalcemia due to hypoparathyroidism (HPT) is a late complication of iron overloaded patients with b-thalassaemia major (TM). The majority of patients have mild disease with parasthesias, while in the more severe form tetany, seizures or cardiac failure may occur. In the last 20 years we observed heart failure in 2 out of 38 (5.2%) TM patients (aged 18 and 22 years) with hypocalcemia secondary to HPT associated to iron overload. Calcium supplementation and vitamin D induced correction of hypocalcemia and resulted in an improvement of cardiac function. Calcium plays a key role in the maintenance and regulation of normal cardiac function. Extra-cellular calcium is indispensable for the contractile process since the sarcoplasmatic reticulum is unable to maintain a sufficient amount of calcium to trigger myocardial contraction. In conclusion, our observations stress the importance of a regular iron chelation therapy, adherence to treatment of endocrine complication and regular follow-up of TM patients with hypocalcemia. PMID- 18403824 TI - Hydrodynamic simulation of multicellular embryo invagination. AB - The mechanical aspects of embryonic morphogenesis have been widely analysed by numerical simulations of invagination in sea urchins and Drosophila gastrulation. Finite element models, which describe the tissue as a continuous medium, lead to the global invagination morphogenesis observed in vivo. Here we develop a simulation of multicellular embryo invagination that allows access to both cellular and multicellular mechanical behaviours of the embryo. In this model, the tissue is composed of adhesive individual cells, in which shape change dynamics is governed by internal acto-myosin forces and the hydrodynamic flow associated with membrane movements. We investigated the minimal structural and force elements sufficient to phenocopy mesoderm invagination. The minimal structures are cell membranes characterized by an acto-myosin cortical tension and connected by apical and basal junctions and an acto-myosin contractile ring connected to the apical junctions. An increase in the apical-cortical surface tension is the only control parameter change required to phenocopy most known multicellular and cellular shape changes of Drosophila gastrulation. Specifically, behaviours observed in vivo, including apical junction movements at the onset of gastrulation, cell elongation and subsequent shortening during invagination, and the development of a dorso-ventral gradient of thickness of the embryo, are predicted by this model as passive mechanical consequences of the genetically controlled increase in the apical surface tension in invaginating mesoderm cells, thus demonstrating the accurate description of structures at both global and single cell scales. PMID- 18403825 TI - Emergent properties during dorsal closure in Drosophila morphogenesis. AB - Dorsal closure is an essential stage of Drosophila development that is a model system for research in morphogenesis and biological physics. Dorsal closure involves an orchestrated interplay between gene expression and cell activities that produce shape changes, exert forces and mediate tissue dynamics. We investigate the dynamics of dorsal closure based on confocal microscopic measurements of cell shortening in living embryos. During the mid-stages of dorsal closure we find that there are fluctuations in the width of the leading edge cells but the time-averaged analysis of measurements indicate that there is essentially no net shortening of cells in the bulk of the leading edge, that contraction predominantly occurs at the canthi as part of the process for zipping together the two leading edges of epidermis and that the rate constant for zipping correlates with the rate of movement of the leading edges. We characterize emergent properties that regulate dorsal closure, i.e., a velocity governor and the coordination and synchronization of tissue dynamics. PMID- 18403826 TI - Dynamical patterning modules: physico-genetic determinants of morphological development and evolution. AB - The shapes and forms of multicellular organisms arise by the generation of new cell states and types and changes in the numbers and rearrangements of the various kinds of cells. While morphogenesis and pattern formation in all animal species are widely recognized to be mediated by the gene products of an evolutionarily conserved 'developmental-genetic toolkit', the link between these molecular players and the physics underlying these processes has been generally ignored. This paper introduces the concept of 'dynamical patterning modules' (DPMs), units consisting of one or more products of the 'toolkit' genes that mobilize physical processes characteristic of chemically and mechanically excitable meso- to macroscopic systems such as cell aggregates: cohesion, viscoelasticity, diffusion, spatiotemporal heterogeneity based on lateral inhibition and multistable and oscillatory dynamics. We suggest that ancient toolkit gene products, most predating the emergence of multicellularity, assumed novel morphogenetic functions due to change in the scale and context inherent to multicellularity. We show that DPMs, acting individually and in concert with each other, constitute a 'pattern language' capable of generating all metazoan body plans and organ forms. The physical dimension of developmental causation implies that multicellular forms during the explosive radiation of animal body plans in the middle Cambrian, approximately 530 million years ago, could have explored an extensive morphospace without concomitant genotypic change or selection for adaptation. The morphologically plastic body plans and organ forms generated by DPMs, and their ontogenetic trajectories, would subsequently have been stabilized and consolidated by natural selection and genetic drift. This perspective also solves the apparent 'molecular homology-analogy paradox', whereby widely divergent modern animal types utilize the same molecular toolkit during development by proposing, in contrast to the Neo-Darwinian principle, that phenotypic disparity early in evolution occurred in advance of, rather than closely tracked, genotypic change. PMID- 18403827 TI - Modeling cell rheology with the Subcellular Element Model. AB - Recently, the Subcellular Element Model (SEM) has been introduced, primarily to compute the dynamics of large numbers of three-dimensional deformable cells in multicellular systems. Within this model framework, each cell is represented by a collection of elastically coupled elements, interacting with one another via short-range potentials, and dynamically updated using over-damped Langevin dynamics. The SEM can also be used to represent a single cell in more detail, by using a larger number of subcellular elements exclusively identified with that cell. We have tested whether, in this context, the SEM yields viscoelastic properties consistent with those measured on single living cells. Employing virtual methods of bulk rheology and microrheology we find that the SEM successfully captures many cellular rheological properties at intermediate time scales and moderate strains, including weak power law rheology. In its simplest guise, the SEM cannot describe long-time/large-strain cell responses. Capturing these cellular properties requires extensions of the SEM which incorporate active cytoskeletal rearrangement. Such extensions will be the subject of a future publication. PMID- 18403828 TI - Mechanical aspects of developmental biology: perspectives on growth and form in the (post)-genomic age. Preface. AB - Simple experiments demonstrate that the development of an organism is both a genetic and a physical process. This statement is so obvious that it is seldom stated explicitly, and yet, there has been little progress toward integrating what should be complementary viewpoints. This paper focuses on the mechanical aspects of morphogenesis-highlighting those areas where mechanics and molecular genetics are converging toward a much-needed synthesis. PMID- 18403829 TI - The forces that shape embryos: physical aspects of convergent extension by cell intercalation. AB - We discuss the physical aspects of the morphogenic process of convergence (narrowing) and extension (lengthening) of tissues by cell intercalation. These movements, often referred to as 'convergent extension', occur in both epithelial and mesenchymal tissues during embryogenesis and organogenesis of invertebrates and vertebrates, and they play large roles in shaping the body plan during development. Our focus is on the presumptive mesodermal and neural tissues of the Xenopus (frog) embryo, tissues for which some physical measurements have been made. We discuss the physical aspects of how polarized cell motility, oriented along future tissue axes, generate the forces that drive oriented cell intercalation and how this intercalation results in convergence and extension or convergence and thickening of the tissue. Our goal is to identify aspects of these morphogenic movements for further biophysical, molecular and cell biological, and modeling studies. PMID- 18403830 TI - Rayleigh instability of the inverted one-cell amphibian embryo. AB - The one-cell amphibian embryo is modeled as a rigid spherical shell containing equal volumes of two immiscible fluids with different densities and viscosities and a surface tension between them. The fluids represent denser yolk in the bottom hemisphere and clearer cytoplasm and the germinal vesicle in the top hemisphere. The unstable equilibrium configuration of the inverted system (the heavier fluid on top) depends on the value of the contact angle. The theoretically calculated normal modes of perturbation and the instability of each mode are in agreement with the results from ComFlo computational fluid dynamic simulations of the same system. The two dominant types of modes of perturbation give rise to axisymmetric and asymmetric sloshing of the cytoplasm of the inverted embryos, respectively. This work quantifies our hypothesis that the axisymmetric mode corresponds to failure of development, and the asymmetric sloshing mode corresponds to development proceeding normally, but with reversed pigmentation, for inverted embryos. PMID- 18403831 TI - Mechanically based generative laws of morphogenesis. AB - A deep (although at the first glance naive) question which may be addressed to embryonic development is why during this process quite definite and accurately reproduced successions of precise and complicated shapes are taking place, or why, in several cases, the result of development is highly precise in spite of an extensive variability of intermediate stages. This problem can be attacked in two different ways. One of them, up to now just slightly employed, is to formulate robust macroscopic generative laws from which the observed successions of shapes could be derived. Another one, which dominates in modern embryology, regards the development as a succession of highly precise 'micropatterns', each of them arising due to the action of specific factors, having, as a rule, nothing in common with each other. We argue that the latter view contradicts a great bulk of firmly established data and gives no satisfactory answers to the main problems of development. Therefore we intend to follow the first way. By doing this, we regard developing embryos as self-organized systems transpierced by feedbacks among which we pay special attention to those linked with mechanical stresses (MS). We formulate a hypothesis of so-called MS hyper-restoration as a common basis for the developmentally important feedback loops. We present a number of examples confirming this hypothesis and use it for reconstructing prolonged chains of developmental events. Finally, we discuss the application of the same set of assumptions to the first steps of egg development and to the internal differentiation of embryonic cells. PMID- 18403832 TI - Laser scanning confocal microscopy coupled with hydraulic permeability measurements for elucidating fluid flow across porous materials: application to human dentine. AB - Laser scanning confocal microscopy (LSCM) coupled to a constant volume flow pressure measuring system is introduced as a new technique for the quantitative measurement of fluid flow across porous materials. Such processes are ubiquitous from the life sciences to materials science and the methodology herein could find widespread application. The methodology has been applied to the detection of fluid flow through human dentine, in-vitro, and in the assessment of occlusion actives. Dentine is a calcareous material sandwiched between the pulp and enamel in the tooth structure that contains tubules which traverse dentine in the pulp to enamel direction. The tubules become patent during enamel erosion or gum recession, leading to dentinal hypersensitivity. Understanding the nature of fluid flow is important, as a pressure gradient exists across dentine in-vivo and this has implications for the development of suitable treatments. The methodology described herein firstly allows a ready assessment of the general efficacy of treatments via hydraulic permeability measurements. Second, LSCM images allow the nature of the flow process and the mode of action of the treatments to be revealed at high spatial resolution. For the particular case of dentine, we demonstrate how the method allows candidate treatments to be compared and assessed. PMID- 18403833 TI - Theoretical treatment of particle-size measurements by applying attenuated-total reflection technique and its comparisons with experimental results. AB - Theoretical equations for a particle size-determination method by applying an attenuated-total-reflection (ATR) technique were obtained. Based on these equations, the ATR-spectral intensities were calculated as functions of the sample amount and on the particle size for various choices of the experimental parameters. A mono layer region was more adequate for size measurements than a multiple-layers region. A prism and the incidence angle should be adequately selected according to the aims of the measurements. Experimental results of the sample amount dependences of the spectral intensities were almost consistent with calculations. The experimental results of size dependences showed the same tendency as those by calculations for SiO2 and SiC; especially for SiC, they were in just accord with those by calculations in the case of a KBr prism. Each component of a mixed plural-kind-particles sample almost agreed in its spectral intensity with that of a corresponding single-kind-particles sample. The particle size-determination method, based on the ATR technique, was theoretically supported, and its characteristics were revealed. PMID- 18403834 TI - An accurate quantitative analysis of polymorphic content by chemometric X-ray powder diffraction. AB - Data from X-ray powder diffraction (XRD) were subjected to a partial least squares regression analysis (PLS) to build a calibration model for predicting the polymorphic content of carbamazepine (CBZ). The effectiveness of the PLS method in the construction of calibration models was analyzed by a scientific approach based on a regulation vector. CBZ forms I and III were characterized by differential scanning calorimetry (DSC) and XRD. Powder mixtures of forms I and III at various ratios (0-100% w/w; form III) were subjected to XRD. Five diffraction peaks were used for the peak-area method to compare with PLS. The results obtained by PLS had a better predictive accuracy compared to those of the peak-area method. The XRD-PLS method was established as a non-destructive, non contact way to avoid the particle orientation effect based on statistical theory. PMID- 18403835 TI - Development of heterodyne detection of dynamic light scattering enhanced by the Talbot effect for the size measurement of nanoparticles. AB - A new type of dynamic light scattering method for the size measurement of nanoparticles was developed using a transmission grating. A sample cell was located behind the grating, and light was incident from the grating side. The scattered light by a solution with nanopariticles was mixed with diffracted light by the grating, and the mixed signal was detected; namely, the diffracted light was used as reference light for heterodyne detection. It was confirmed that the S/N ratio of the autocorrelation curve was 26-times improved by heterodyne detection. Furthermore, the S/N ratio was improved by setting the sample cell at the sample grating distance where the electromagnetic field is maximum due to the Talbot effect. Size measurements for several kinds of nanoparticles were demonstrated by this new method. PMID- 18403836 TI - Molecular design of tetraazamacrocyclic derivatives bearing a spirobenzopyran and three carboxymethyl moieties and their metal-ion complexing behavior. AB - Tetraazacyclododecane and tetraazacyclotetradecane derivatives bearing a spirobenzopyran and three carboxymethyl moieties, 1 and 2, and a diethylenetriamine derivative bearing a spirobenzopyran and four carboxymethyl moieties 3 were synthesized. The isomerization behaviors based on the spirobenzopyran moiety of these ligands were studied by UV-visible spectrophotometry in aqueous solutions containing various metal ions at neutral pH. These ligands formed stable 1:1 complexes with lanthanide ions, while the spirobenzopyran moiety was isomerized to its corresponding merocyanine form even under dark conditions. In aqueous solutions containing a lanthanide ion, the absorption spectra of 1 or 2 showed remarkable blue shifts, while absorbances at the maximum absorption wavelengths in the visible region were enhanced; such changes are attributable to the isomerization to the merocyanine form of the spirobenzopyran moiety. These results suggest that the phenolate anion of the merocyanine moiety interacts very strongly with a lanthanide ion bound by the complexing moiety because of the high charge density of lanthanide ions. In contrast, the absorbance of merocyanine form was decreased by the complexation of the macrocyclic ligand with transition metal ions, such as Cu2+ and Zn2+. This result indicates that macrocyclic ligands, 1 and 2, formed complexes with transition metal ions only by the aminocarboxylate moieties, and the phenolate ion of merocyanine moiety was not able to participate in the complexation. This conclusion was also demonstrated by density functional theory calculations. PMID- 18403837 TI - Facile assay of telomerase activity utilizing a DNA-detectable chemiluminogenic reagent. AB - Telomerase shows increased activity in most human cancers and germ line cells, but not in normal human somatic cells. We describe a novel chemiluminescence method for the facile assay of telomerase activity in human cells. The telomerase substrate was incubated with the cell lysate containing various amounts of telomerase, and then the telomerase product was amplified by the polymerase chained reaction (PCR). The PCR products were separated from the excess substrate, primer and deoxyribonucleotide triphosphates by a centrifugal filter, which distinguished different molecular sizes. The isolated products were reacted with a DNA-detectable chemiluminogenic reagent, 3,4,5-trimethoxyphenylglyoxal. The proposed assay method gave linearity for the telomerase activity in 100 to 10000 cells (r2=0.997), and allowed the assay not only of lower activity, but also of higher activity of telomerase without the requirement of any special labeled-PCR primers in the assay system. PMID- 18403838 TI - Direct determination of horseradish peroxidase encapsulated in liposomes by using luminol chemiluminescence. AB - Horseradish peroxidase (HRP) encapsulated in liposomes was directly detected by using luminol chemiluminescence (CL) with H2O2 without lysis of liposomes. At a low concentration of H2O2, the initial rate of HRP-catalyzed luminol CL in liposomes was slower than that of HRP-catalyzed luminol CL in a lipid-free bulk solution. The decrease in the initial rate of the CL reaction in liposomes was due to the membrane permeation of luminol and H2O2. At a high concentration of H2O2, the initial rate of the CL reaction in liposomes was the same as that in a lipid-free bulk solution. The CL measurement conditions in both a lipid-free bulk solution and in liposomes were optimized in the concentrations of luminol and H2O2 by measuring the CL response curves, in which only one peak appeared and the CL intensity was maximal. The CL intensity observed in HRP-catalyzed luminol CL in liposomes was a factor of seven greater than that observed in a lipid-free bulk solution. The CL intensity was dependent on the amount of HRP-encapsulated liposomes used. The detection limit in the direct detection of HRP encapsulated in liposomes was sensitive by a factor of 3 compared with that in HRP-catalyzed luminol CL in a lipid-free bulk solution. PMID- 18403839 TI - Amperometric biosensor based on glucose dehydrogenase and plasma-polymerized thin films. AB - A novel design is described for an amperometric biosensor based on NAD(P) dependent glucose dehydrogenase (GDH) combined with a plasma-polymerized thin film (PPF). The GDH is sandwiched between several nanometer thick acetonitrile PPFs on a sputtered gold electrode (PPF/GDH/PPF/Au). The lower PPF layer plays the role as an interface between enzyme and electrode because it is extremely thin, adheres well to the substrate (electrode), has a flat surface and a highly crosslinked network structure, and is hydrophilic in nature. The upper PPF layer (overcoating) was directly deposited on immobilized GDH. The optimized amperometric biosensor characteristics covered 2.5-26 mM glucose concentration at +0.6 V of applied potential; the least-squares slope was 320 nA mM(-1) cm(-2) and the correlation coefficient was 0.990. Unlike conventional wet-chemical processes that are incompatible with mass production techniques, this dry-chemistry procedure has great potential for enabling high-throughput production of bioelectronic devices. PMID- 18403840 TI - Amperometric biosensor for hydrogen peroxide based on direct electrocatalysis by hemoglobin immobilized on gold nanoparticles/1,6-diaminohexane modified glassy carbon electrode. AB - A facile strategy of an amperometric biosensor for hydrogen peroxide based on the direct electrocatalysis of hemoglobin (Hb) immobilized on gold nanoparticles (GNPs)/1,6-diaminohexane (DAH) modified glassy carbon electrode (GCE) has been described. A uniform monolayer film of DAH was initially covalently bound on a GCE surface by virtue of the electrooxidation of one amino group of DAH, and another amino group was modified with GNPs and Hb, successively. The fabrication process was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). The proposed biosensor exhibited an effective and fast catalytic response to the reduction of H2O2 with good reproducibility and stability. A linear relationship existed between the catalytic current and the H2O2 concentration in the range of 1.5x10(-6) to 2.1x10(-3) M with a correlation coefficient of 0.998 (n=24). The detection limit (S/N=3) was 8.8x10(-7) M. PMID- 18403841 TI - Electrochemical analysis of chloramphenicol using boron-doped diamond electrode applied to a flow-injection system. AB - The electrochemical properties of chloramphenicol at a boron-doped diamond thin film (BDD) electrode were studied using cyclic voltammetry. The highest current response of chloramphenicol was obtained with phosphate buffer, pH 6 (0.1 M) in 1% ethanol. The relationship between the concentration of chloramphenicol and the current response was linear over the range of 0.1-10 mM (R2=0.9990). The amount of chloramphenicol was analyzed by flow-injection analysis. A thin-layer flow cell equipped with a BDD electrode was used as an amperometric detector, and experiments were carried out at -0.7 V (vs. Ag/AgCl). The linear relationship between the current response and the concentration of chloramphenicol in the range of 0.1-50 microM (R2=0.9948) and the limit of detection of 0.03 microM (S/N=3) were obtained. This method has been successfully applied to the determination of chloramphenicol in sterile eye drops and milk sample by the standard addition method. The average recoveries of chloramphenicol in eye drops were 98.0%, and the average recoveries of chloramphenicol from spiked milk were 93.9-103%. PMID- 18403842 TI - Development and validation of a stability-indicating micellar electrokinetic chromatography method for the determination of ezetimibe in pharmaceutical formulations. AB - A micellar electrokinetic chromatography (MEKC) method was validated for the analysis of ezetimibe. The method was carried out on a fused-silica capillary (50 microm i.d.; effective length, 40 cm). The background electrolyte consisted of a 25 mM borate buffer and 25 mM anionic detergent SDS (pH 9.75)/methanol (90:10, v/v). The capillary temperature was maintained at 35 degrees C, the applied voltage was 30 kV; the injection was performed using a pressure mode at 50 mbar for 5 s, with detection at 232 nm. The method was linear in the range of 2-150 microg/mL (R2=0.9999). The specificity and the stability-indicating capability were proven through degradation studies, which also showed that there was no interference of the excipients. The limits of quantitation and detection were 2 and 0.41 microg/mL, respectively. The method was applied for the analysis of ezetimibe pharmaceutical formulations, and the results were compared to those of the liquid-chromatography method. PMID- 18403843 TI - Spectrophotometric determination of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride by flow injection analysis. AB - 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCl) is a very useful agent to form amide bonds (peptide bonds) in an aqueous medium. A simple and fast detection system was developed using the reaction with pyridine and ethylenediamine in acidic aqueous solution and spectrophotometric flow injection analysis. The absorbances were measured at 400 nm and the reaction was accelerated at 40 degrees C. The calibration graph showed good linearity from 0 to 10% of EDC.HCl solutions: the regression equation was y=3.15x10(4)x (y, peak area; x, % concentration of EDC.HCl). The RSD was under 1.0%. Sample throughput was 15 h(-1). This method was applied to monitoring the EDC.HCl concentration that remained after the anhydration of phthalic acid in water, esterification of acetic acid in methanol or dehydration condensation of malonic acid and ethylenediamine in water. PMID- 18403844 TI - Determination of iodine and bromine compounds by ion chromatography/dynamic reaction cell inductively coupled plasma mass spectrometry. AB - An inductively coupled plasma mass spectrometer (ICP-MS) was used as an ion chromatographic detector for the speciation of iodine and bromine. Gradient elution using NH4NO3 at pH 10 allowed the chromatographic separation of ionic iodine (I- and IO3-) and bromine (Br- and BrO3-) species in less than 8 min. Effluents from the ion-exchange column were delivered to the nebulization system of ICP-MS for the determination of I and Br. The potentially interfering 38Ar40ArH+ and 40Ar40ArH+ at the bromine masses m/z 79 and 81 were significantly reduced in intensity (by approximately two orders of magnitude) by using 0.6 mL min(-1) O2 as a reactive cell gas in the dynamic reaction cell (DRC). Moreover, the signal-to-background ratio at iodine mass m/z 127 increased significantly when O2 was used as the reaction gas. The detection limits were in the range of 0.001-0.002 and 0.03-0.04 ng mL(-1) for various I and Br compounds, respectively, based on the peak height. The relative standard deviation of the peak areas for five injections of a 2 ng mL(-1) I-, IO3- and 20 ng mL(-1) Br-, BrO3- mixture was in the range of 3-4%. The concentrations of I and Br compounds have been determined in selected water and urine samples. The spike recoveries were in the range of 94-102% for all of the determinations. This method has also been applied to determine various I and Br compounds in an NIST RM 8435 whole-milk powder reference material and a seaweed sample obtained locally. A microwave-assisted extraction method was used to extract these compounds, which were quantitatively leached with a 10% mass/volume (m/v) tetramethylammonium hydroxide (TMAH) solution in a focused microwave field within a period of 6 min. The major components of I and Br in milk powder and seaweed were I- and Br-. PMID- 18403845 TI - Multi-layer cartridges filled with multi-walled carbon nanotubes for the determination of volatile organic compounds in indoor air. AB - A kind of new multi-layer adsorbent including Tenax TA/multi-walled carbon nanotubes (MWCNTs)/Carboxen 564 was developed for collecting volatile organic compounds. The adsorption and desorption efficiencies of 11 kinds of adsorbents (including Activated charcoal, Tenax TA, Carboxen 564, Chromosorb 101, Chromosorb 102, Chromosorb 103, Chromosorb 105, Porapak Q, GDX 301, XAD-2, and MWCNTs) were compared. By combining the advantages of Tenax TA, MWCNTs, and Carboxen 564, new type of multi-layer adsorbents was developed. The adsorption and desorption efficiency, the sampling reproducibility, and the effect of water were improved using multi-layer adsorbents, Tenax TA/MWCNTs/Carboxen 564. New multi-layer adsorbents were successfully applied to the determination of volatile organic compounds (VOCs) in ambient air. PMID- 18403846 TI - Accurate measurement of atmospheric helium isotopes. AB - We have developed an analytical system to measure atmospheric 3He/4He ratios precisely by using a static vacuum operation mass spectrometer and an ultrahigh vacuum purification line. After purification of the sample, helium was introduced into the mass spectrometer. Ion beams of 3He and 4He were measured at the same time by a double collector system. A resolving power of about 700 at the 5% level of the peak height was attained for the complete separation of 3He ions from those of HD and H3. Repeated analysis of a sample calibrated against a standard showed a precision of about 0.2% error margin (2sigma). This system has been applied to evaluating the 3He/4He ratio of He Standard of Japan (HESJ), and for the assessment of the atmospheric variation of latitude. The observed 3He/4He ratio of HESJ, 20.405+/-0.040Rair (2sigma) agrees well with the value of 20.408+/ 0.044Rair by Lupton and Evans, but is smaller than the recommended value of 20.63+/-0.10Rair in a literature. Air samples collected from 45 to 20 degrees N in the Far East showed a slight decrease of the 3He/4He ratio with latitude. Even though the variation is attributable to an experimental artifact, it may not preclude the possibility that the anthropogenic release of crustal 4He is significant in the high-latitude region of the northern hemisphere. PMID- 18403847 TI - Application of laser-induced photoacoustic spectroscopy for determination of plutonium concentration in nuclear waste solutions. AB - Laser-induced photoacoustic spectroscopy was used in a quantitative analysis of Pu in HNO3 medium. Plutonium was quantitatively oxidized to Pu(VI) using Ce(IV). The photoacoustic measurement of Pu(VI) with maximum absorption at 830.5 nm was subsequently performed to determine the concentration. The photoacoustic signal was linearly proportional to the Pu(VI) ion concentration. The detection limit of Pu(VI) was estimated to be 0.5 microg mL(-1) (3sigma) in 3 M HNO3. By the proposed method, Pu concentration was successfully determined in a nuclear waste solution for use in nuclear materials management. PMID- 18403848 TI - Selective pressurized liquid extraction of polychlorinated biphenyls in sediment. AB - A selective pressurized liquid extraction procedure (SPLE) was developed for a fast determination of polychlorinated biphenyls in sediment. The final method was performed at 100 degrees C with heptane/dichloromethane (90:10, v/v) as extraction solvent for 2x5 min. Sulfuric acid impregnated silica was placed downstream of the sample in the extraction cell to remove interfering components. This simultaneous extraction/clean-up was performed in 20 min, with an average congener recovery of 92% compared to a classical 24 h Soxhlet methodology and 2 h of external manual clean-up. PMID- 18403849 TI - Selective determination of diphenhydramine in compound pharmaceutical containing ephedrine by flow-injection electrochemiluminescence. AB - On the basis of the structural interactions between diphenhydramine (DPH) and ephedrine (EPH) to enhance electrochemiluminescence (ECL) intensity, a flow injection ECL analysis of DPH in the present of EPH by utilizing tris(2,2' bipyridine)ruthenium(II), (Ru(bpy)(3)2+) has been reported. In the optimized experimental conditions, the linear ECL response to concentrations of DPH is from 2.00 to 40.0 microg/L with a correlation coefficient of 0.9995 and a detection limit of 1.20 microg/L. The relative standard deviation (RSD) was less than 4.6% (n=5) and the recovery was in the range of 98-106% for the determination of DPH in pharmaceutical samples. The method avoids any interference of coexistent EPH in the compounding drug without prior separation. The method has advantages over HPLC method in terms of speed and convenience, economics and safe procedure, and could be an alternative for places where HPLC equipment is not available. PMID- 18403850 TI - Analysis of red phosphorus in resins using pyrolysis-gas chromatography/mass spectrometry. AB - Although red phosphorus is used as a flame retardant for polymer materials, no analysis methods for it in resins has been established. Analysis methods for red phosphorus in resins were investigated using pyrolysis-gas chromatograpy/mass spectrometry (Py-GC/MS) by paying attention to the fact that it has a sublimation property. We found that the mass spectrum of red phosphorus shows a series of ions at m/z=62, 93 and 124, and a fragment pattern indicating that red phosphorus (P4=124, P=31) was pyrolytically decomposed. The coefficient of the correlation between the content of red phosphorus in the resin and the peak intensity in Py GC/MS was 0.9781. The relative standard deviations of this analysis method was 6.29% (n=5). Py-GC/MS was applicable not only to qualitative analysis but also to the quantitative analysis of red phosphorus. PMID- 18403851 TI - pH-potentiometirc study of polyion complexes between chitosan and poly(vinyl sulfate) or dodecylbenzene sulfonate. AB - Polyion complexes of three chitosans with poly(vinyl sulfate) (PVS) and dodecylbenzene sulfonate (DBS) were examined by a potentiometric study that was to separately measure the pH of sample solutions individually prepared. Apparent formation constants (Ki) of ion association between the protonated amines of chitosan and the sulfates of PVS or the sulfonates of DBS were determined. The effects of pH, coexistent salt concentration, and molecular weight on the values of Ki were investigated in order to reveal the properties of the complexation. The values of Ki for chitosan-PVS were quite larger than that for chitosan-DBS. The deducing effect of the coexistent salt was strong against chitosan-PVS, but was weak against chitosan-DBS. Thus, chitosan-PVS complexes possessed a strong electrostatic binding, and chitosan-DBS complexes included a hydrophobic interaction. For chitosan-PVS complexes the effect of the coexistent salt was weaker for a high molecular weight of chitosan than for a low molecular weight. PMID- 18403852 TI - Development of an automatic phase-contrast microscopic system capable of determining the microbial density and distribution inside an immobilized carrier. AB - The cell-immobilization technique is used for the physical and chemical fixation of cells onto a solid support in order to increase their stability and capacity of substrate uptake. However, there is no apparatus to observe the microbial community's structure inside cell immobilizing polymeric carrier. In order to satisfy the demand of monitoring for the microbial distribution inside the carrier, we developed an automatic phase contrast microscopic monitoring system capable of determining the microbial density and distribution inside a cell entrapped carrier automatically. PMID- 18403854 TI - Short history of epidemiology for noninfectious diseases in Japan. Part 3: nutritional dystrophia. PMID- 18403855 TI - Dietary patterns and levels of blood pressure and serum lipids in a Japanese population. AB - BACKGROUND: Associations between dietary patterns and cardiovascular disease risk factors remain unclear. The objective of this study was to evaluate the association between dietary patterns derived from factor analysis and the levels of blood pressure and serum lipids in a Japanese population. METHODS: We conducted a cross-sectional analysis among 6886 (in the analysis on blood pressure) and 7641 (in the analysis on serum lipids) Japanese subjects aged 40-69 years. Dietary patterns were identified from a food frequency questionnaire by factor analysis. Associations between dietary patterns and blood pressure and serum lipids were examined after taking potential confounders into account. RESULTS: Three dietary patterns were identified: vegetable, meat, and Western. In men, the meat pattern was associated with higher total, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) cholesterol. The Western pattern was associated with higher total and LDL cholesterol. In women, the vegetable pattern was associated with lower systolic and diastolic blood pressure and pulse pressure, and higher HDL cholesterol. The meat pattern was associated with higher total and HDL cholesterol. The Western pattern was associated with higher total, HDL, and LDL cholesterol, and the least intake pattern of Western diet was associated with higher systolic and diastolic blood pressures. CONCLUSIONS: Dietary patterns of a Japanese population were related to cardiovascular disease risk factors, especially in women. PMID- 18403856 TI - Predictive validity of the total health index for all-cause mortality assessed in the Komo-Ise cohort. AB - BACKGROUND: The Total Health Index (THI), a self-administered questionnaire developed in Japan, is used for symptom assessment and stress management of employees and others; however, it has not been reported whether it can predict mortality risk. METHODS: The THI, with 12 primary and 5 secondary scales, was applied to a cohort consisting of middle-aged residents in Japan. This study, called the Komo-Ise cohort study, was started in 1993. The scale scores were related to 481 deaths from all causes among 10,816 residents over 93 months. The statistics were tested by the Cox hazard model and adjusted for three background variables (sex, age, and district where the subject resided). RESULTS: Five of the scales [depression and aggression (primary scales), and psychosomatics, neurotics, and schizophrenics (secondary scales)] indicated significant hazard ratios for mortality. The lowest quintile group of the aggression scale score had the largest hazard ratio of 2.58, compared with the middle quintile group (95% confidence interval: 1.88-3.52). The psychosomatics, neurotic scales and depression scales also had a minimum hazard ratio in the middle quintile group. One of the secondary scales, T1, which represents a somatoform disorder, had a significant linear relationship with the mortality risk, although its proportionality with the cumulative mortality rates was not satisfactory. CONCLUSIONS: Five scales of the THI were significantly related to mortality risk in the Komo-Ise cohort, which could be used for score evaluation and in the personal health advice system of the THI. PMID- 18403858 TI - A six-month follow-up study of maternal anxiety and depressive symptoms among Japanese. AB - BACKGROUND: Maternal psychological distress has been widely studied, but epidemiologic data based on follow-up studies of maternal psychological distress remain insufficient in Japan. The objective of this study was to estimate the prevalence of anxiety and depressive symptoms among child-rearing women in Japan at two time-points after childbirth. METHODS: A self-administered questionnaire was delivered on two occasions to 2,657 women who had given birth in 2004: first when their infants were 3-4 months old and then again when their infants were 9 10 months old. The questionnaire included the Hospital Anxiety and Depression Scale (HADS; Japanese version) to estimate the level of maternal psychological distress. RESULTS: The total percentage of women with anxiety symptoms as assessed by a HADS score of 8+ was 26.2 % at 3-4 months of age, and 26.1 % at 9 10 months. Among the women without anxiety symptoms at 3-4 months, 11.6 % showed anxiety symptoms at 9-10 months. The total percentage of depressive symptoms was 19.0 % at 3-4 months, and 24.0 % at 9-10 months. Among the women without depressive symptoms at 3-4 months, 14.0 % showed depressive symptoms at 9-10 months. CONCLUSION: Anxiety symptoms in mothers appeared to persist from 3-4 months to 9-10 months after childbirth, while depressive symptoms tended to be more common at 9-10 months after childbirth. Nevertheless, the prevalence of anxiety symptoms was higher than that of depressive symptoms. PMID- 18403859 TI - Mortality from dermal exposure to tetramethylammonium hydroxide. PMID- 18403857 TI - Active smoking, passive smoking, and breast cancer risk: findings from the Japan Collaborative Cohort Study for Evaluation of Cancer Risk. AB - BACKGROUND: Evidence is lacking regarding the relationship between cigarette smoking and breast cancer in Japanese women. We examined the association between breast cancer incidence and active and passive smoking in the Japan Collaborative Cohort Study for Evaluation of Cancer Risk. METHODS: Our study comprised 34,401 women aged 40-79 years who had not been diagnosed previously with breast cancer and who provided information on smoking status at baseline (1988-1990). The subjects were followed from enrollment until December 31, 2001. Cox proportional hazards models were used to estimate the hazard ratio (HR) and 95% confidence interval (CI) for the association between breast cancer incidence and tobacco smoke. RESULTS: During 271,412 person-years of follow-up, we identified 208 incident cases of breast cancer. Active smoking did not increase the risk of breast cancer, with a HR for current smokers of 0.67 (95% CI: 0.32-1.38). Furthermore, an increased risk of breast cancer was not observed in current smokers who smoked a greater number of cigarettes each day. Overall, passive smoking at home or in public spaces was also not associated with an increased risk of breast cancer among nonsmokers. Women who reported passive smoking during childhood had a statistically insignificant increase in risk (HR: 1.24; 95% CI: 0.84-1.85), compared with those who had not been exposed during this time. CONCLUSION: Smoking may not be associated with an increased risk of breast cancer in this cohort of Japanese women. PMID- 18403860 TI - Patterns of outpatient visits by Japanese male expatriates in Thailand. AB - Objective of this study is to clarify the health-related conditions of Japanese male expatriates in Thailand. Records of Japanese male expatriates in their 30s to 50s who consulted outpatient clinics at Bangkok Hospital in 2005 were analyzed for number and type of outpatient visits then compared with data from the "Patient Survey" 2005 for age-matched males in Japan. "Diseases of the respiratory system" and "Certain infectious and parasitic diseases" had a higher rate of diagnosis among Japanese men living in Thailand than those living in Japan. "Endocrine, nutritional and metabolic diseases," and "Mental and behavioral disorders" had a lower rate of diagnosis among Japanese men living in Thailand than those in Japan. "Acute upper respiratory infections" accounted for 87.4% of "Diseases of the respiratory system" and "intestinal infections" accounted for 59.9% of "Certain infectious and parasitic diseases" among expatriates. Japanese male expatriates have a tendency of not visiting hospital when they have no subjective or objective symptoms. To support a healthy expatriate life, health information should be propagated widely and available support services, such as counseling via e-mail or fax, should be actively promoted. Expatriates should be informed of the availability of these services before they go abroad and during their stay. PMID- 18403861 TI - Characterization of liver injury associated with hypersensitive skin reactions induced by trichloroethylene in the guinea pig maximization test. AB - Trichloroethylene (TCE) can induce non-dose-related hepatitis, possibly classified as delayed-type hypersensitivity (immune-mediated hepatitis), as well as dose-related toxic liver injury. However, the difference in pathophysiology between the two kinds of hepatitis remains unknown. This study aimed to characterize the liver injury associated with hypersensitive skin reactions induced by TCE in guinea pigs. As a model of dose-related acute toxic liver injury, the animals were treated with intradermal injection (ii) (0, 167, 500, 1500 or 4500 mg/kg of TCE) or dermal patch (dp) (0 or 900 mg/kg of TCE). The guinea pig maximization test (GPMT) was also carried out as a model of immune mediated liver injury, in which the total TCE dosage was below 340 mg/kg. In the group of TCE 4500 mg/kg (ii), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) increased (p<0.01), while total protein and globulin decreased (p<0.05). Evident fatty degeneration, hepatic sinusoid dilation and inflammatory cell infiltration were observed. No significant change was found in animals treated with TCE of doses below 500 mg/kg (ii) or 900 mg/kg (dp). In the GPMT, sensitization rates of TCE-induced dermal allergy were 66%. ALT, AST, lactate dehydrogenase and the relative liver weight increased significantly (p<0.05) while albumin, IgA and gamma-glutamyl transpeptidase decreased significantly (p<0.05). Lesions of ballooning changes were observed in liver pathology. Thus, TCE could cause both acute-type toxic liver injury and immune mediated liver injury, the so-called delayed-type hypersensitivity at doses below the dosage for toxic liver injury. Interestingly, the histopathological features were quite different: fatty degeneration was most prominent in the former, and ballooning in the latter. PMID- 18403862 TI - Development of a sampling and analysis method for 4-vinyl-1-cyclohexene in air. AB - The purpose of this research was to develop an applicable sampling and analytical method to determine airborne 4-vinyl-1-cyclohexene concentrations which are usually found in the atmosphere of polybutadiene factories. A solid sorbent tube, containing two sections (100 mg in the front and 50 mg in the back) of activated coconut-shell charcoal was chosen for sampling 4-vinyl-1-cyclohexene vapor. The 4 vinyl-1-cyclohexene in the charcoal samples was desorbed with carbon disulfide and analyzed by gas chromatography equipped with a flame ionization detector. The suitable air flow rate, adsorption capacity, sample storage stability, desorption efficiency and reliability of the method for sampling and analysis of 4-vinyl-1 cyclohexene were evaluated. The method was applied to sampling and analysis of 4 vinyl-1-cyclohexene in the rubber industry. The results indicated a suitable air flow rate of 0.3 to 1.5 l/min. The adsorption capacity of 4-vinyl-1-cyclohexene on 100 mg of charcoal was 0.2134 mg. The 4-vinyl-1-cyclohexene adsorbed on the charcoal was stable for 7 d at room temperature or 21 d in a refrigerated condition. The average percent desorption efficiency of 4-vinyl-1-cyclohexene ranged from 90.45% to 97.04% with the loaded amount ranging from 0.412 to 8.250 microg using 1 ml carbon disulfide. The limit of detection of 4-vinyl-1 cyclohexene was 0.044 ng. The average percent recoveries (n=6) of 4-vinyl-1 cyclohexene adsorbed on charcoal ranging from 0.46 to 8.87 microg were 96.78 102.87% with relative standard deviations (RSDs) of 0.34-1.92%, respectively. The concentrations of 4-vinyl-1-cyclohexene ranged from 0.011 to 0.105 mg/m(3) in the working environment of a polybutadiene factory. PMID- 18403863 TI - Asthma mortality in male workers of the dye industry in Korea. AB - Workers in the dye industry are exposed to various chemicals, of which reactive dye is a well-known occupational asthmagen. This study examined the relationship between asthma mortality and occupational exposure in the dye industry. The cohort comprised 66,089 male workers, including 904 workers in the dye industry, who underwent medical examinations from 1995 to 2003 at a medical service institute located in Incheon, Korea. Deaths were also observed during the 1995 to 2003 period. The mortality was analyzed using the standardized mortality ratio (SMR) compared to the Korean general population, and the mortalities from asthma in dye manufacturing workers and other workers were compared using the standardized rate ratio (SRR). The all-cause mortality in dye industry workers was significantly lower than in the general population (SMR=0.40, 95% CI 0.24 0.63), while the asthma mortality (SMR=9.03, 95% CI 1.86-26.39) was significantly higher. Deaths from non-malignant respiratory diseases were higher in dye industry workers, but were not statistically significant (SMR=2.0, 95% CI 0.41 5.86). In the internal comparison, the all-cause mortality was the same in both groups, while the mortalities from non-malignant respiratory diseases (SRR=6.04, 95% CI 4.00-9.44) and asthma (SRR=23.29, 95% CI 11.05-58.83) were higher in dye industry workers than in workers in other industries. Because asthma is a life threatening disease, special consideration and preventive measures must be taken for workers in the dye industry. PMID- 18403864 TI - An occupational health study of emergency physicians in Japan: health assessment by immune variables (CD4, CD8, CD56, and NK cell activity) at the beginning of work. AB - This study was conducted to evaluate the occupational health of Japanese physicians in emergency medicine. Subjects participating in this study were eighty-nine physicians working at 12 medical facilities (10 critical care emergency centers) in Japan. Participants were asked to complete a questionnaire of work conditions and to provide blood samples for immune variable measurements (CD4, CD8, CD56 and natural killer cell (NK cell) activity) before commencing their work. The data collected from seventy-four of 89 participating physicians were analyzed. The traditional work group comprised of 39 emergency physicians, who were significantly overworked compared to other two groups: the shift work group and the day work group. Among these three groups, no immune variable was significantly different except lymphocyte, number of CD4, and NK cell activity; and the NK cell activity of the shift work group was significantly lower than those of the traditional work group (p<0.01) and the day work group (p<0.01) in terms of Bonferroni's multiple comparison, probably due to circadian rhythm. It was indicated that NK cell activity was significantly lower in samples collected at night versus in the morning (OR=8.34, 95%CI: 1.95-35.6, p<0.01) through multiple logistic regression analyses. NK cell activity was significantly lower in individuals taking 0-3 days off per month, as compared to those taking 4 or more days off (OR=4.65, 95%CI: 1.27-17.0, p=0.02), according to multiple logistic regression analyses. Therefore, the low NK cell activity appears to have reflected the extent of fatigue arising from physicians' overwork. Overwork would have been a potential risk for the physicians' health, resulting in a lower quality of Japanese emergency medical services than that which could have been achieved otherwise. This study suggests that it would be better for the Japanese emergency physicians to take 4 or more days off per month for their health and the quality of their services. PMID- 18403865 TI - Selenium-enriched Japanese radish sprouts influence glutathione peroxidase and glutathione S-transferase in an organ-specific manner in rats. AB - Selenium-enriched Japanese radish sprouts (Se-enriched JRS), in which Se methylselenocysteine accounted for 80% of Se compounds, inhibited mammary tumorigenesis induced by 7,12-dimethylbenz[a]anthracene in rats. The effects of Se-enriched JRS on the oxidative stress-scavenging enzymes were investigated in rats. F344 female rats were fed test diets, in which Se-enriched JRS was added at 0, 2.4, 5.0, 8.8 or 12.5 ppm Se to commercial rodent chow for 3 wk. Glutathione peroxidase (GPx) and glutathione S-transferase (GST) in rat livers, kidneys and lungs were measured. Tissue Se concentrations at the highest Se dose (12.5 ppm) were high in order as follows: kidney > liver > lung. The diet at 12.5 ppm Se reduced the increase in body weight and, conversely, increased the liver weight. The Se test diets decreased hepatic and renal GPx activity at more than 2.4 ppm and 5.0 ppm, respectively. In contrast, the test diets increased pulmonary GPx activity at more than 2.4 ppm Se. The diets increased hepatic GST activity at more than 2.4 ppm Se dose dependently, whereas they reduced pulmonary GST activity at more than 2.4 ppm. The diet of 12.5 ppm Se induced GST Yp in all 3 organs and GST Yb1 in the liver. Thus, Se-enriched JRS influenced GPx and GST activity in a symmetrical manner in the livers and lungs of rats, with hepatic GST possibly affected, in part, by the induction of GST Yb1. PMID- 18403866 TI - Psychosocial work environment and well-being: a cross-sectional study at a thermal power plant in China. AB - To investigate the effects of the job demand-control (DC) model and the effort reward imbalance (ERI) model on worker's well-being, self-reports for psychosocial work conditions and well-being were made by a sample of 878 workers at a thermal power plant in China using the main dimensions of DC and ERI questionnaires. Logistic regression analyses were employed controlling for age, gender, and educational level, and negative and positive affection among others. Workers reporting high job demands and low job control or high efforts and low rewards had elevated risks of job dissatisfaction, psychosomatic complaints and depressive symptoms. Odds ratios were generally higher in workers reporting both high efforts and low rewards. Furthermore, low reward proved to be a stronger predictor of poor well-being when both job stress models were simultaneously adjusted. To some extent, interaction effects were found for social support, but no interaction effects were found for overcommitment. The findings indicate independent effects of both the DC model and the ERI model on well-being. Future work should explore the combined effects of these two models of psychosocial stress at work on health more thoroughly. PMID- 18403867 TI - Musculoskeletal disorder among 52,261 Chinese restaurant cooks cohort: result from the National Health Insurance Data. AB - To investigate the incidence of musculoskeletal disorders (MSDs) and the particular MSDs to which Chinese restaurant cooks are most at risk in Taiwan, National Health Insurance Data from 1998 to 2002 were used to identify MSDs for study populations, including 52,261 certified cooks and 209,044 references matched for age and sex. The annual incidence of MSDs was around 25% and 20% for cooks and references, respectively, and the cook-to-reference risk ratios ranged from 1.29 to 1.35 (p<0.001). The most frequently affected body part was the low back, but the epicondyle was at higher risk (OR>2) than other sites for the cooks. Cooks are at higher risk of having MSDs complaints and MSDs related to the elbow show the highest risk. PMID- 18403868 TI - Different mechanisms of DEHP-induced hepatocellular adenoma tumorigenesis in wild type and Ppar alpha-null mice. AB - Di (2-ethylhexyl) phthalate (DEHP) exposure is thought to lead to hepatocellular hypertrophy and hyperplasia in rodents mediated via peroxisome proliferator activated receptor alpha (PPAR alpha). A recent study revealed that long-term exposure to relatively low-dose DEHP (0.05%) caused liver tumors including hepatocellular carcinomas, hepatocellular adenomas, and chologiocellular carcinomas at a higher incidence in Ppar alpha-null mice (25.8%) than in wild type mice (10.0%). Using tissues with hepatocellular adenoma, microarray (Affymetrix MOE430A) as well as, in part, real-time quantitative PCR analysis was conducted to elucidate the mechanisms of the adenoma formation resulting from DEHP exposure in both genotyped mice. The microarray profiles showed that the up- or down-regulated genes were quite different between hepatocellular adenoma tissues of wild-type and Ppar alpha-null mice exposed to DEHP. The gene expressions of apoptotic peptidase activating factor 1 (Apaf1) and DNA-damage inducible 45 alpha (Gadd45a) were increased in the hepatocellular adenoma tissues of wild-type mice exposed to DEHP, whereas they were unchanged in corresponding tissues of Ppar alpha-null mice. On the other hand, the expressions of cyclin B2 and myeloid cell leukemia sequence 1 were increased only in the hepatocellular adenoma tissues of Ppar alpha-null mice. Taken together, DEHP may induce hepatocellular adenomas, in part, via suppression of G2/M arrest regulated by Gadd45a and caspase 3-dependent apoptosis in Ppar alpha-null mice, but these genes may not be involved in tumorigenesis in the wild-type mice. In contrast, the expression level of Met was notably increased in the liver adenoma tissue of wild-type mice, which may suggest the involvement of Met in DEHP-induced tumorigenesis in wild-type mice. PMID- 18403869 TI - Working condition factors associated with time pressure of nurses in Japanese hospitals. AB - In Japan, the most common reason for medical errors is a lack of cross-checking. To prevent errors, efforts to strengthen cross-checking behaviors are being adopted. However, time pressures also lead to errors, and increasing cross checking activities leads to an increased workload and even greater time pressures. The purpose of this study was to identify working conditions that lead to time pressure for nurses, and to find ways to reduce time pressure and prevent subsequent errors. Self-reporting questionnaires were distributed over 10 days to 416 nurses working in 17 wards at two hospitals; 357 nurses (85.8%) responded anonymously, providing data on 2,150 person-days. In multivariate analyses, medical support services and the number of nurse calls answered were associated with subjective assessments of time pressures and nursing service delays due to busyness. Moreover, working the "evening-day shift" (when a nurse works a day shift after working the evening shift with no days off in between) led to a high level of fatigue before work and was associated with nursing service delays due to busyness. Reducing time pressures and preventing errors requires an adequate number of nurses, shift plans that consider rest periods and order of rotation, increased task discretion for nurses, and the prevention of chronic fatigue. PMID- 18403870 TI - Effects of web-based career identity training for stress management among Japanese nurses: a randomized control trial. PMID- 18403871 TI - Earplug-type earphone with built-in microphone improves monosyllable intelligibility in noisy environments. PMID- 18403872 TI - A case of occupational allergic contact dermatitis due to PVC hose. PMID- 18403873 TI - Workplace violence directed at nursing staff at a general hospital in southern Thailand. AB - This study aimed to document the characteristics of workplace violence directed at nursing staff, an issue which has rarely been studied in a developing country. Two study methods, a survey and a key informant interview, were conducted at a general hospital in southern Thailand. A total of 545 out of 594 questionnaires sent were returned for statistical analysis (response rate=91.7%). The 12-month prevalence of violence experience was 38.9% for verbal abuse, 3.1% for physical abuse, and 0.7% for sexual harassment. Psychological consequences including poor relationships with colleagues and family members were the major concerns. Patients and their relatives were the main perpetrators in verbal and physical abuse while co-workers were the main perpetrators in cases of sexual harassment. Common factors to incidents of violence were psychological setting, illness of the perpetrators, miscommunication, and alcohol use. Logistic regression analysis showed younger age to be a personal risk factor. Working in the out-patient unit, trauma and emergency unit, operating room, or medical or surgical unit increased the odds of violence by 80%. Training related to violence prevention and control was found to be effective and decreased the risk of being a victim of violence by 40%. We recommend providing training to high risk groups as a means of controlling workplace violence directed at nursing staff. PMID- 18403874 TI - High prevalence of diffuse parenchymal lung diseases among Turkish tinners. AB - The objective is to investigate the prevalence of parenchymal lung diseases among tinners. Twenty-four tinners who signed an informed consent form in the Gaziantep area were the subjects of this study. Demographics, spirometric test results, and high resolution computed tomography (HRCT) scans were obtained for all the tinners. Out of 24 tinners, eleven tinners (46%) had HRCTs consistent with DPLD. Of these 11 subjects, HRCT findings were consistent with respiratory bronchiolitis interstitial lung disease in nine, usual interstitial pneumonia pattern in one, and non-specific interstitial pneumonia in one. HRCT patterns consistent with DPLDs are significantly prevalent among Turkish tinners. This is the first field study reporting this occupational lung hazard in tinners. Future research is needed to uncover the underlying mechanisms. PMID- 18403875 TI - Association between use of synthetic metalworking fluid and risk of developing rhinitis-related symptoms in an automotive ring manufacturing plant. AB - The main objective of this study was to determine the association between synthetic metalworking fluid (MWF) and rhinitis-related symptoms. At a plant manufacturing piston rings for automobiles, we interviewed grinders (19) and manufacturing workers (142) in operations where synthetic or semisynthetic MWF is handled, and administrative office workers (44) regarding the principal symptoms of rhinitis (nasal stuffiness, runny nose, anosmia, nasal itchiness, rhinorrhea, headache, epistaxis, and post-nasal drip). In addition, we assessed the current exposure of workers handling MWF to MWF aerosols, fungi, and endotoxins. Logistic regression analysis was used to examine the association between MWF surrogates indicative of MWF exposure and each rhinitis-related nasal symptom. Odds ratios (ORs) and 95% confidence intervals were adjusted for sex, age, smoking habit, and duration of employment. Among grinders handling synthetic MWF, the frequency of complaints of the dominant symptoms was 66.7% for nasal stuffiness, 77.8% for anosmia, 77.8% for runny nose, and 50.0% for headache. These rates are quite high even allowing for the common occurrence of rhinitis in the general population. Twenty eight of 34 grinding and manufacturing workers (82.4%) sampled were exposed to MWF mist above the threshold limit of 0.2 mg/m(3) listed as a notice of intended change by the American Conference for Governmental Industrial Hygienists (ACGIH). The percentage of workers exposed to MWF mist >0.5 mg/m(3) was 17.6%. Most workers were exposed to fungi levels >103 CFU/m(3). All exposures to endotoxins were <50 EU/m(3). Logistic regression analysis found that use of synthetic MWF was significantly associated with excess risk of nasal stuffiness (OR 3.5), nasal itchiness (OR 2.0), and runny nose (OR 2.1). The use of semi synthetic MWF had little or no impact on the risk of developing rhinitis-related nasal symptoms. Grinding workers handling synthetic MWF had an increased risk of nasal stuffiness (OR 7.9), anosmia (OR 23.2), nasal itchiness (OR 8.3), runny nose (OR 20.4), post nasal drip (OR 18.4), and headache (OR 7.4) compared to administrative workers. Synthetic MWF may play an important role in the development of the dominant symptoms of rhinitis. Further study is needed to establish the risk of rhinitis or rhinitis-related symptoms according to MWF type. PMID- 18403876 TI - A pilot study to evaluate the adhesion of oral microorganisms to temporary soft lining materials. AB - The aim of the study was to compare the adhesion of oral microorganisms to different types of soft liner and acrylic resin surfaces. Three different soft lining materials were applied to cavities formed on the fitting surfaces of prostheses in 17 patients. On days 1, 7 and 14, the specimens were taken out and immediately processed for bacteriological evaluation. The numbers of adhering microorganisms were calculated and the specimens were compared among each other and also with a control group (acrylic resin). Data were analyzed by two-way ANOVA and least squares differences at a significance level of P < 0.05. Among the four materials tested the total number of oral microorganisms adhering to Softliner material was the greatest after each of the time periods tested. Higher numbers of oral bacteria and Candida were shown to adhere to soft lining materials than to acrylic resin. Microbial coverage increased continuously with time and the differences between days 1 and 14 were statistically significant (P < 0.05). Temporary soft lining materials are not resistant to adhesion and possible surface damage caused by oral bacteria, and therefore their use should be limited to short-term periods. PMID- 18403877 TI - Root canal configuration and the prevalence of C-shaped canals in mandibular second molars in an Iranian population. AB - A total of 139 extracted mandibular second molars were injected with India ink and demineralized. They were made clear and transparent with methyl salicylate, and the anatomy of their canals was studied. It was found that 86.3% of mandibular second molars had two roots, 9.3% had one root, and 4.3% had three roots. Ninety percent of the mesial roots of the mandibular second molars with two roots had two canals (predominantly with a type II or III configuration), and 77.5% of the distal roots of these teeth had one canal (predominantly with a type I configuration). Among the mandibular second molars, 7.2% had C-shaped canals and these configurations were seen mostly in single-rooted mandibular second molars. The results of this study indicate that mandibular second molar teeth have many variations in the number of roots and the morphology of their canals. Therefore it should not be assumed that all mandibular second molar teeth have two roots and three canals. PMID- 18403878 TI - Prevalence and distribution of odontogenic cysts in Sicily: 1986-2005. AB - The objective of this study was to assess the prevalence of odontogenic cysts (OCs) in Sicily and evaluate their distribution during a 20-year period. A cross sectional retrospective study was carried out in 1,310 cysts of the jaw diagnosed in 12,197 individuals, who consecutively attended the Odontostomatologic Clinic of Palermo from 1986 to 2005. 1,273 cysts were classified as odontogenic, whereas only 37 were non-odontogenic. In the former group, the most frequent lesions were radicular cysts (84.5%), followed by dentigerous OCs (11.4%). Inflammatory radicular cysts were observed more in male gender, younger age at diagnosis and anterior maxilla as site of presentation. Unlike dentigerous cysts, the frequency of radicular cysts decreased from 10.4% in 1986-1995 to about 8% in 1996-2005 (P < 0.0001). Inflammatory radicular cysts are the most represented group among OCs in our area with a higher prevalence than that reported in other countries. The decreasing epidemiologic trend with time could be attributed to the possible changes in individual oral health behavior. PMID- 18403879 TI - Effects of use of anabolic steroids on the masticatory system: a pilot study. AB - The use of androgenic anabolic steroids (AAS) has increased significantly among athletes in Brazil and other countries. These drugs alter the physiological behavior of bone and muscles, also affecting these structures in masticatory system. This paper aims to evaluate bone and dental changes in users of AAS, as well as the incidence of temporomandibular dysfunction (TMD), compared to athletes not using AAS. Eight athletes were equally divided in two groups, AAS users and non-users. The groups were evaluated using Helkimo index, McNamara cephalometric tracing and cast analysis. The AAS users presented more intense TMD signs and symptoms (Di total value, P = 0.096, Mann-Whitney test), increased cephalometric measures (Co-A, P = 0.020, Mann-Whitney test) and Angle Class II malocclusion, compared to the non-users. These results suggested that the use of AAS alters masticatory structures and increases the incidence of TMD. PMID- 18403880 TI - Detection of Epstein-Barr virus and human cytomegalovirus in blood and oral samples: comparison of three sampling methods. AB - The purpose of this study was to identify and compare the presence of HCMV and EBV-1 in subgingival plaque, unstimulated saliva and peripheral blood of patients with chronic periodontitis. Forty patients diagnosed with chronic periodontitis (mean age, 41.7 years) were recruited. Unstimulated saliva, subgingival plaque and peripheral blood were collected from each patient and the DNA of each sample was isolated. The viruses were detected using the nested PCR technique. The detection frequency of EBV-1 in subgingival plaque, saliva and peripheral blood was 45%, 37.5% and 25%, respectively. HCMV was detected in 82.5% of subgingival plaque samples and peripheral blood and in 75% of salivary samples. The sensitivity for detecting EBV-1 in saliva and peripheral blood when EBV-1 was detected in subgingival plaque samples was low (22% and 27.7%, respectively) and the sensitivity for detecting HCMV in saliva and peripheral blood when compared to subgingival plaque was high (81.8% and 87.8%, respectively). There is a high agreement among the three sampling methods in detection of HCMV, but the detection of EBV-1 would require a combination of saliva and subgingival plaque sampling to avoid false negative results. PMID- 18403881 TI - Angina bullosa hemorrhagica of the soft palate: a clinical study of 16 cases. AB - Angina bullosa hemorrhagica (ABH) is an oral mucosal blood blister that develops without blood dyscrasia or vesiculobullous disorder. Although a minor mucosal trauma has been suggested as a triggering factor for ABH, its etiopathogenesis, especially the causative role of systemic conditions, is largely unknown. We investigated the presence or absence of local factors as well as systemic background disease in 16 patients with ABH arising in the soft palate. All the lesions were solitary, and 75% of them (n = 12) appeared during the ingestion of hard or crispy food. With regard to underlying systemic conditions, hypertension was the most common (n = 6), and asthma, insomnia, diabetes mellitus, rheumatoid arthritis, gastrointestinal disorder and hyperuricemia were also recorded (n = 1 each). Five patients had no significant background disease. There were no recalcitrant or recurrent cases. In conclusion, the present study has revealed that scratching of the oral mucosa during eating plays an important role in the formation of ABH. Hypertension appears to be the most frequent background condition, but its pathogenic relationship with ABH remains speculative, as hypertension is fairly common in adults. PMID- 18403882 TI - Influence of light intensity on contraction stress of flowable resins. AB - The purpose of this study was to evaluate the influence of power density on contraction stress of resin composite restorative materials during photo polymerization. Six flowable resin composites, and a hybrid resin composite for comparison, were used. The composites were polymerized with the power density adjusted to either 100 or 600 mW/cm(2). Stress development was determined with a custom-made tensilometer. The adhesive was placed in a thin layer on a steel rod and resin paste was packed into the mold. The contraction force (N) generated during polymerization was continuously recorded and the maximum contraction stress (MPa) was calculated. Data were analyzed statistically. When the power density was adjusted to 100 mW/cm(2), the average contraction stress ranged from 0.30 to 0.50 MPa for the flowable composites, compared with 0.35 MPa for the hybrid composite. When the power density was adjusted to 600 mW/cm(2), the average contraction stress ranged from 0.34 to 1.00 MPa for the flowable composites and 0.69 MPa for the hybrid composite comparison. For all materials tested except Estelite Flow Quick, contraction stress increased with higher power density. The present results indicate that contraction stress during polymerization is influenced by power density and resin composite type. PMID- 18403883 TI - The effects of PNF training on the facial profile. AB - Although orthodontic treatment improves dentoalveolar problems, the facial profile seldom changes because the perioral muscles do not easily adapt to the new morphological circumstances. We employed proprioceptive neuromuscular facilitation (PNF), which is training with added resisted movement to motions such as lifting the upper lip, lowering the lower lip, and sticking out the tongue, to adapt the perioral muscles to the new morphological circumstances. The subjects were 40 adults with an average age of 29.6 years. A series of PNF exercises was performed three times per day for 1 month. Lateral facial photographs were taken using a digital camera before training (T(0)), after training (T(1)), and 1 month after the end of training (T(2)). The nasolabial (NL), mentolabial (ML), and mentocervical (MC) angles were measured, and linear measurements were taken to verify the change of each measurement point. In the test group, the NL and ML angles significantly increased (P < 0.05), and the MC angle significantly decreased after the PNF exercise. From T(1) to T(2), the NL and ML angles decreased significantly, while the MC angle increased significantly. No significant differences were observed in these angles when the values measured at T(0) and T(2) were compared. Although the training appeared to be effective for sharpening the mouth and submandibular region, continued training is necessary to avoid relapse. PMID- 18403884 TI - Biomarkers of periodontitis in oral fluids. AB - Matrix metalloproteinases (MMPs), the key enzymes responsible for matrix degradation, are derived from polymorphonuclear leukocytes during the early stages of periodontitis. The present study determined the levels of GCF matrix metalloproteinase-2 (MMP-2) and metalloproteinase-9 (MMP-9) and salivary MMP-8 in patients with gingivitis and periodontitis and in healthy controls. Levels of crevicular MMP-2, MMP-9 and salivary MMP-8 were determined by ELISA in subjects with healthy gingiva (n = 15), gingivitis (n = 18) and periodontitis (n = 20). Significantly higher salivary MMP-8 and crevicular MMP-9 were observed in cases of periodontitis compared to gingivitis and healthy adults. On the other hand, crevicular MMP-2 levels in periodontitis subjects were lower than those in gingivitis and healthy subjects. The three MMP levels were highly correlated to probing depth, and bleeding on probing. Salivary MMP-8, crevicular MMP- 2 and 9 may serve as biomarkers of periodontal disease and aid in early detection of periodontitis or gingivitis. PMID- 18403885 TI - Effect of chewing a mixture of areca nut and tobacco on periodontal tissues and oral hygiene status. AB - The present study was conducted to clarify the effects of chewing a quid containing areca nut and tobacco on periodontal tissue and oral hygiene status. A total of 365 subjects (168 chewers and 197 non-chewers with a mean age of 32.5 +/ 0.7 and 30.4 +/- 0.8 years, respectively) were enrolled. Clinical data on periodontal tissues, oral hygiene status, as well as information on bleeding from gums, ulcers in the oral cavity, or a burning sensation in the soft tissues, were collected as indicators of the possible presence and extent of periodontal lesions. The results indicated that a significantly higher number of quid-chewers suffered bleeding from the gums, halitosis, difficulty in opening the mouth and swallowing solid food, a burning sensation in the soft tissues, and ulcers in the oral cavity than non-chewers. There was no significant difference between quid chewers and non-chewers with respect to oral hygiene measures adopted. However, clinical examination using the oral hygiene index score indicated that the oral hygiene status of quid-chewers was significantly deteriorated. The effect of quid chewing on the periodontium, i.e. the occurrence of periodontal pockets, gingival lesions and gum recession, were significantly higher in quid-chewers than in non chewers. Age, sex and smoking adjusted odds ratios for quid-chewers against non chewers using logistic regression analysis indicated that, in general, chewers were at significantly higher risk for various oral complaints and periodontium status. The present data indicate that chewing quid comprising areca nut and tobacco has adverse effects on periodontal tissues, oral hygiene and incidence of oral lesions. PMID- 18403886 TI - Residual antibacterial activity of chlorhexidine and MTAD in human root dentin in vitro. AB - The purpose of this in vitro study was to compare the antimicrobial substantivity of BioPure MTAD, 2% chlorhexidine (CHX) and 2.6% sodium hypochlorite (NaOCl) in human root dentin. One hundred and ten dentin tubes prepared from human maxillary incisors were infected in vitro for 14 days with Enterococcus faecalis. The specimens were divided into five groups as follows: CHX; BioPure MTAD; NaOCl; infected dentin tubes (positive control); and sterile dentin tubes (negative control). Dentin chips were collected with round burs into Brain Heart Infusion (BHI) broth. After culturing, the number of colony-forming units (CFU) was counted. In all experimental groups, CFU was minimum after treatment (day 0), and the results obtained were significantly different from each other at any time period (P < 0.05). After treatment, the NaOCI group and BioPure MTAD group showed the lowest and highest number of CFU, respectively. In each group, the number of CFUs increased significantly by time-lapse (P < 0.05). In conclusion, the substantivity of BioPure MTAD was significantly greater than CHX and NaOCl. PMID- 18403887 TI - Study of root canal accessibility in human primary molars. AB - The aim of the present study was to provide a general scheme for pulpectomy of primary molars that may be useful for decision-making about negotiation of root canals and selection of appropriate instruments. A total of 160 vital primary molars in 85 patients (40 males, 45 females) aged 4-6 years were selected. After taking primary radiographs, local anesthesia was induced, and the teeth were isolated using a rubber dam. Canal accessibility index (CAI) and tooth accessibility index (TAI) were calculated according to initial file size. Mandibular first molars had either three canals (79.2%) or four canals (20.8%), and all second molars had four canals. Maxillary first molars had three canals and second molars had either three canals (70.9%) or four canals (29.1%). Lower accessibility of the mandibular first molar distobuccal root accounted for the lower accessibility of these teeth in comparison with mandibular second molars. While three-canal maxillary second molars were more accessible due to the lower accessibility of the distobuccal canal of the maxillary first molar, poor accessibility of the distal canal in four-canal second molars was responsible for the difficult accessibility of these teeth. In conclusion, it seems that the accessibility of a single canal in each tooth determines the difficulty of accessibility for any given tooth. Moreover, while primary second molars are more accessible than first molars, all of them are negotiable. PMID- 18403888 TI - Effects of Smads and BMPs induced by Ga-Al-As laser irradiation on calcification ability of human dental pulp cells. AB - We investigated the effects of Ga-Al-As laser irradiation on the mineralization ability of human dental pulp (HDP) cells and on Smads and bone morphogenetic protein (BMP) production as one mechanism for the transmission of laser photochemical energy to cells. HDP cells in vitro were irradiated once with a Ga Al-As laser at 1.0 W for 500 s, and calcified nodule formation was assessed by Alizarin red S staining. The laser irradiation was greater in the laser irradiated group than in the non-irradiated group. Both calcium production and alkaline phosphatase (ALP) activity were higher after laser irradiation. Expression of mRNAs for Smad1, Smad7, BMPs, ALP, and osteocalcin was greater after laser irradiation, whereas expression of Smad6 mRNA was inhibited. Production of BMP-2 and BMP-4 in conditioned medium was also higher after laser irradiation. These results suggest that Smads and BMPs play important roles in ALP activity and calcification upon laser irradiation of HDP cells. PMID- 18403889 TI - Effects of interleukin-1alpha on the production and release of basic fibroblast growth factor in cultured nifedipine-reactive gingival fibroblasts. AB - The effect of interleukin-1alpha (IL-1alpha) on the production of basic fibroblast growth factor (bFGF) in human gingival fibroblasts originated from a nifedipine-reactive patient was investigated. Ca(2+)-mobilizing agents, thapsigargin and bradykinin, were also tested to determine whether they affected the production and release of bFGF. The release of bFGF from IL-1alpha-pretreated cells in relation to the transient increase in intracellular Ca(2+)([Ca(2+)]i) and extracellular Ca(2+)levels was also investigated. IL-1alpha and thapsigargin yielded significantly higher bFGF production, and also enhanced bFGF mRNA expression. IL-1alpha-pretreated cells showed significantly greater release of bFGF under the present experimental conditions. Levels of released bFGF were significantly higher in cells pretreated with IL-1alpha, followed by bradykinin and thapsigargin in the presence of extracellular Ca(2+). The transient mobilization of intracellular Ca(2+) accelerated the release of bFGF in IL-1alpha pretreated cells, but not in untreated cells. Thus, IL-1alpha increases bFGF production in nifedipine-reactive gingival fibroblasts and also influences the release of bFGF in the IL-1alpha-pretreated cells. PMID- 18403890 TI - Intraosseous verrucous carcinoma originating from an odontogenic cyst: a case report. AB - Carcinoma originating in bone is uncommon; most of them apparently arise in odontogenic cysts. In this paper, we report an extremely rare case in which, verrucous carcinoma originated from an odontogenic cyst. This lesion was firm and non-tender on palpation and had a white-pink appearance. It was encased in the anterior aspect of the maxilla and exhibited microscopic features of verrucous carcinoma of the oral mucosa. After surgical enucleation, no recurrence or metastasis has been observed up to now. It is mandatory to correlate the clinical and histopathologic findings to establish a true diagnosis. PMID- 18403891 TI - Primary intraosseous adenoid cystic carcinoma of the mandible with lung metastasis: a case report. AB - Intraosseous adenoid cystic carcinoma is an extremely rare neoplasm with only 17 cases reported previously. A case of primary intraosseous adenoid cystic carcinoma of the mandible with lung metastasis in a 45-year-old man is reported, together with a brief review of the literature. PMID- 18403892 TI - Endodontic treatment of a radiculous maxillary premolar: a case report. AB - We report the diagnosis and successful clinical management of three patients with anatomical variation in the maxillary first premolars. Maxillary first premolars have highly variable root canal morphology, but the presence of three canals is rare. This article describes the diagnosis and clinical management of first maxillary premolars with three canals and three separate roots, with special reference to radiographic interpretation and access refinements. PMID- 18403893 TI - Oral and maxillofacial myiasis associated with epidermoid carcinoma: a case report. AB - Myiasis is a term derived from the Greek word "myia", meaning invasion of vital tissue of humans or other mammals by fly larvae. The deposited eggs develop into larvae, which penetrate deep structures causing adjacent tissue destruction. It is an uncommon clinical condition, being more frequent in underdeveloped countries and hot climate regions, and is associated with poor hygiene, suppurative oral lesions, alcoholism and senility. Its diagnosis is made basically by the presence of larvae. This paper reports a case of oral and maxillofacial myiasis involving 273 larvae in a patient with epidermoid carcinoma without physical or neurological deficiency. The patient's management was antisepsis, larval removal and general care, before death after three months. PMID- 18403894 TI - Favorable response of an extensive periapical lesion to root canal treatment. AB - This article presents non-surgical resolution of an extensive periapical lesion of endodontic origin associated with the maxillary left lateral incisor. Clinical examination revealed an asymptomatic bony hard swelling confined to the palate, while radiographic analysis showed a lesion measuring 22 mm in diameter and nearly 389 mm(2) in area. Through apical patency, 4 ml of intracanal exudate was drained. After thorough biomechanical preparation, a calcium hydroxide/CPMC root canal dressing was applied and periodically renewed for 11 months. The exudate was eliminated at treatment onset and significant bone formation was observed at the periapical region in the following months with concomitant resolution of the cortical expansion. Complete radiographic resolution of the periapical lesion was observed two years after the root canal filling. Thus, non-surgical treatment of this supposedly cystic, extensive periapical lesion provided favorable clinical and radiographic response. PMID- 18403895 TI - Fabrication of a maxillary posterior fixed partial denture with a type 4 gold alloy and a dual-polymerizing indirect composite. AB - The aim of the present study was to design and fabricate a maxillary posterior fixed partial denture (FPD) made of a type 4 gold alloy and an indirect composite. Unlike the conventional framework design of a resin veneered restoration, cut-back was extended approximately 1/4 to 1/3 width in the occlusal table of the buccal cusp. Multiple retentive beads 150-200 mum in diameter were placed on the metal surface to be veneered. The gold alloy was cast in a cristobalite mold using a centrifugal casting machine. The cut surface with the retentive beads was air-abraded with alumina, and a priming agent (Alloy Primer) that contained triazine dithione monomer (VTD) was applied. A tooth-colored veneer was then fabricated with a highly loaded light- and heat-cured composite material (Estenia). This design and procedure can be applied as a standardized laboratory technique for fabrication of maxillary posterior restorations and FPDs. PMID- 18403896 TI - Decreased expression of angiotensin II type 1 and type 2 receptors in the brain after long-term administration of antihypertensive drugs in stroke-prone spontaneously hypertensive rat. AB - The present study examined the levels of Angiotensin II type 1 receptor (AT(1)) and type 2 receptor (AT(2)) in the brain stem and cerebral cortex of the stroke prone spontaneously hypertensive rat (SHR-sp) after long-term treatment with three types of antihypertensive drugs: valsartan, enalapril, and amlodipine. In both tissues, expression of the AT(1) was decreased by administration of each drug. Expression of the AT(2) was decreased in the cerebral cortex by drug administration, but did not change in the brain stem. This study may contribute to elucidating the relationship between AT(1) and AT(2) expressions and the effect of antihypertensive drugs in SHR-sp brain. PMID- 18403897 TI - Oseltamivir enhances hippocampal network synchronization. AB - Oseltamivir, a widely used anti-influenza drug, inhibits virus neuraminidase. A mammalian homologue of this enzyme is expressed in the brain, yet the effect of oseltamivir on central neurons is largely unknown. Patch-clamp recordings ex vivo revealed that oseltamivir enhanced spike synchronization between hippocampal CA3 pyramidal cells. Time-lapse multineuron calcium imaging revealed that oseltamivir and its active metabolite evoked synchronized population bursts that recruited virtually all neurons in the network. This unique, so-far-unknown, event was attenuated by muscarinic receptor antagonist. Thus, oseltamivir is a useful tool for investigating a new aspect of neural circuit operation. PMID- 18403898 TI - Selegilin exerts antidepressant-like effects during the forced swim test in adrenocorticotropic hormone-treated rats. AB - In the present study, we investigated the effect of adrenocorticotropic hormone (ACTH) on the immobilization of rats in the forced swim test after the administration of selegiline, a selective and irreversible monoamine oxidase (MAO)-B inhibitor. Single and repeated administration of selegiline significantly decreased the duration of immobility in normal rats. When selegiline was administered for 15 days, we observed a significant decrease in immobility in rats treated with ACTH for 14 days. The immobility-decreasing effect of selegiline was blocked by nafadotride, a selective dopamine D(3)-receptor antagonist in normal and ACTH-treated rats. Selegiline may be useful in an animal model of depressive conditions resistant to tricyclic antidepressant treatment via the dopamine D(3) receptor. PMID- 18403899 TI - Effect of aripiprazole on 5-HT2 receptor-mediated wet-dog shake responses and disruption of prepulse inhibition in rats. AB - Aripiprazole, an atypical antipsychotic drug, is a D(2) dopamine-receptor partial agonist, but also has affinity to several serotonin receptors (5-HT(1A,2A,2C,7)). However, little is known about the contribution of serotonin receptors in the action of aripiprazole. The present study investigated the effects of aripiprazole on 5-HT(2A) receptor-mediated behaviors and compared them with the effects on dopamine receptor-mediated behavior in rats. Aripiprazole (10, 30 mg/kg, p.o.) inhibited the stereotyped behavior induced by apomorphine (1 mg/kg, s.c.), a dopamine-receptor agonist, and the wet-dog shake responses induced by DOI (2,5-dimethoxy-4-iodoamphetamine, 2 mg/kg, s.c.), a 5-HT(2A)-receptor agonist. Moreover, aripiprazole improved the disruption of prepulse inhibition induced by both apomorphine and DOI significantly. These data suggest that not only the dopaminergic system, but also the serotonergic system are involved in the antipsychotic effect of aripiprazole. PMID- 18403900 TI - Effects of rosuvastatin and pitavastatin on ischemia-induced myocardial stunning in dogs. AB - Incomplete recovery of myocardial contraction after reperfusion following brief ischemia is called the "stunning phenomenon" in an animal experiment. A hydrophilic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor (statin) does not affect this phenomenon, but lipophilic statins further reduce the contraction during reperfusion. The effects of novel hydrophilic rosuvastatin and lipophilic pitavastatin on myocardial stunning in dogs were examined. In a preliminary experiment in vitro, pitavastatin reduced L6 cell viability at 10(-6) M and higher, whereas rosuvastatin and pravastatin up to 10(-5) M did not show such effects. An empty capsule or a capsule filled with rosuvastatin (2 mg/kg per day) or pitavastatin (0.4 mg/kg per day) was orally administered to dogs. After 3 weeks, both statins lowered the serum cholesterol level to the same extent. Under pentobarbital anesthesia, dogs were subjected to 15-min ischemia followed by 120 min reperfusion. Ischemia arrested the myocardial contraction in the ischemic area, and reperfusion recovered it but incompletely, showing the stunning phenomenon. Rosuvastatin did not modify the stunning phenomenon, while pitavastatin further deteriorated the myocardial contraction during reperfusion. PMID- 18403901 TI - Evidence for separate involvement of different mu-opioid receptor subtypes in itch and analgesia induced by supraspinal action of opioids. AB - The common adverse effect of centrally-injected mu-opioid receptor (mu-OR) agonists is pruritus. This study was conducted using mice to examine whether different subtypes of mu-OR would be responsible for pruritus and analgesia. Intracisternal injections of morphine and morphine-6beta-glucronide (M6G), but not M3G, produced an antinociceptive effect. Morphine, but neither M6G nor M3G, induced facial scratching, a pruritus-related response. Facial scratching following morphine was not affected by the mu(1)-OR antagonist naloxonazine at doses that inhibited the antinociceptive effects. The results suggest that different subtype and/or splice variants of mu-OR are separately involved in pruritus and antinociception of opioids. PMID- 18403902 TI - Analysis of cell cycle in mouse bone marrow cells following acute in vivo exposure to 56Fe ions. AB - A pilot study was conducted to examine the magnitude of cell-cycle delay and apoptosis in bone marrow (BM) cells collected at 18, 42 and 66 hr from radiosensitive CBA/CaJ mice and radioresistant C57BL/6J mice following a whole body in vivo exposure to 1 GeV/amu (56)Fe ions or (137)Cs gamma rays. At each sacrifice, BM cells were collected from three mice of each strain per dose of (56)Fe ions (0, 10 and 100 cGy) and two mice of each strain per dose of (137)Cs gamma rays (0, 100 and 300 cGy). A significant G1-arrest (ANOVA, p < 0.05) was observed at 18 hr after exposure of mice to 100 cGy of (56)Fe ions or 300 cGy of (137)Cs gamma rays, relative to their corresponding sham-controls, resulting in a significant decrease in the percentage of cells cycling into S-phase in both strains. The percentage of S-phase cells subsequently increased and persisted up to 66 hr post-irradiation. Significant numbers of G2/M cells were found at 18 and 66 (but not at 42) hr post-irradiation, regardless of radiation-type or mouse strain. It is likely that BM cells have undergone at least one cell cycle at 66 hr after exposure of mice to either 100 cGy (56)Fe ions or 300 cGy (137)Cs gamma rays. Our study is the first to investigate the in vivo effects of (56)Fe ions (1 GeV/amu) on the cell cycle of mouse BM cells using flow cytometry. The cell-cycle distribution (but not the number of apoptotic cells) was dependent on radiation dose and harvest-time. PMID- 18403903 TI - ShRNA-mediated Ku80 gene silencing inhibits cell proliferation and sensitizes to gamma-radiation and mitomycin C-induced apoptosis in esophageal squamous cell carcinoma lines. AB - To investigate the effects of Ku80 depletion on cell growth and sensitization to gamma-radiation and MMC-induced apoptosis in esophageal squamous cell carcinoma lines. Six human carcinoma cell lines (LNcaP, K562, MDA-MB-231, MCF-7, EC9706, and K150) and normal HEK293 cell line were examined for basal levels of Ku80 protein by western blotting analysis. The suppression of Ku80 expression was performed using vector-based shRNA in EC9706 cells. Cell proliferation was determined with MTT assay and colony formation assay and tumorigenicity in a xenograft model in vitro and in vivo. Sensitivity of EC9706 cells treated with shRNA vector to gamma-radiation and MMC was determined with colony formation assay and MTT assay. The cell cycle distribution was determined by Flow cytometry. Apoptosis induced by gamma-radiation and MMC was analyzed using GENMED TUNEL FACS kit. Ku80 showed higher basal levels in six carcinoma cell lines than in HEK293. The suppression of Ku80 expression decreased cellular proliferation, colony formation and inhibited tumorigenicity in a xenograft model. Furthermore, it sensitized apoptosis of the cancer cells induced by gamma-radiation and MMC. Ku80 plays an important role not only in tumorigenesis but also in radiation resistance and chemotherapy resistance in esophageal cancer cells. Hence Ku80 may serve as a promising therapeutic target, particularly for recurrent esophageal tumors. PMID- 18403913 TI - Regulation of adipose tissue metabolism in cancer cachexia. AB - PURPOSE OF REVIEW: Body-fat depletion is a hallmark of cancer cachexia, a complex clinical syndrome associated with increased morbidity and mortality. Understanding the fat-loss disorder in cachexia is essential for the development of better treatments for the syndrome. This review presents recent studies focusing on the mechanisms of adipose atrophy in cancer cachexia, particularly the potential mediators. RECENT FINDINGS: Fat loss cannot be explained by poor appetite alone, and may also result from altered lipid metabolism in adipocytes. Increased lipolysis appears to be a key factor underlying fat loss in cancer cachexia though decreases in lipid deposition and adipocyte development may also contribute. Both tumour and host-derived factors are implicated in adipose tissue atrophy. Tumour necrosis factor-alpha has been associated with increased lipolysis in adipocytes. The novel adipokine zinc-alpha2-glycoprotein may function locally, as well as systemically, to promote lipid mobilization and utilization in cancer cachexia. SUMMARY: Clarifying the role of cachexia mediators in adipose tissue atrophy will add to our understanding of adipocyte metabolism in wasting disease. Elucidating their mode of action may lead to novel therapeutic targets for counteracting the cachexia syndrome. PMID- 18403914 TI - The dermcidin gene in cancer: role in cachexia, carcinogenesis and tumour cell survival. AB - PURPOSE OF REVIEW: The diverse protein products of the dermcidin gene are relevant to immunity, cancer cell progression and cancer cachexia. This article evaluates recent developments/controversies around dermcidin. RECENT FINDINGS: Dermcidin has recently been shown to act as a survival/proliferation factor in hepatoma and prostate cancer cell lines. Recent studies suggest that the Y-P30 subunit of the dermcidin polypeptide offers a survival advantage in such cancer cells. Nevertheless, the relevance of Y-P30 to cancer growth in vivo, and mechanisms of action remain unknown. In mice, tumour cells appear to glycosylate the Y-P30 subunit, transforming it into a potent skeletal muscle proteolysis inducing factor. Recent work has described a receptor and signal transduction pathways for murine glycosylated proteolysis-inducing factor. The absence of classical N-glycosylation sites in the human proteolysis-inducing factor peptide and the lack of specific tools for the detection of the key carbohydrate moieties conferring the proteolysis-inducing activity, however, remain barriers to confirming glycosylated proteolysis-inducing factor as a pro-cachectic factor in humans. SUMMARY: There is a growing body of evidence illustrating dermcidin as an oncogene and Y-P30 as a survival factor. The biology of murine proteolysis inducing factor as a pro-cachectic factor continues to evolve; however, its role in human biology remains speculative. PMID- 18403915 TI - Computational modeling of cancer cachexia. AB - PURPOSE OF REVIEW: Measurements of whole-body energy expenditure, body composition, and in-vivo metabolic fluxes are required to quantitatively understand involuntary weight loss in cancer cachexia. Such studies are rare because cancer cachexia occurs near the end of life when invasive metabolic tests may be precluded. Thus, models of cancer-associated weight loss are an important tool for helping to understand this debilitating condition. RECENT FINDINGS: A computational model of human macronutrient metabolism was recently developed that simulates the normal metabolic adaptations to semi-starvation and re-feeding. Here, this model was used to integrate data on the metabolic changes in patients with cancer cachexia. The resulting computer simulations show how the known metabolic disturbances synergize with reduced energy intake to result in a progressive loss of body weight, fat mass, and fat-free mass. The model was also used to simulate the effects of nutritional support and investigate inhibition of lipolysis versus proteolysis as potential therapeutic approaches for cancer cachexia. SUMMARY: Computational modeling is a new tool that can integrate clinical data on the metabolic changes in cancer cachexia and provide a conceptual framework to help understand involuntary weight loss and predict the effects of potential therapies. PMID- 18403916 TI - Leucine-enriched nutrients and the regulation of mammalian target of rapamycin signalling and human skeletal muscle protein synthesis. AB - PURPOSE OF REVIEW: To highlight recent studies that have examined the cell signalling mechanisms responsible for the amino acid (primarily leucine and the essential amino acids) stimulation of human skeletal muscle protein synthesis. RECENT FINDINGS: Ingestion of a leucine-enriched essential amino acid nutrient solution rapidly and potently activates the mammalian target of rapamycin signalling pathway and protein synthesis in human skeletal muscle. Further, mTOR signalling and muscle protein synthesis are enhanced when leucine-enriched nutrients are ingested following resistance exercise. The addition of leucine to regular meals may improve the ability of feeding to stimulate protein synthesis in old human muscle. SUMMARY: Leucine and essential amino acids appear to stimulate human muscle protein synthesis primarily by activating the mammalian target of rapamycin signalling pathway. How human muscle cells sense an increase in leucine and/or essential amino acids to activate mammalian target of rapamycin signalling is currently unknown. Recent work, however, suggests that the kinases hVps34 and MAP43K may be involved. Leucine-enriched essential amino acid ingestion, in combination with resistance exercise in some cases, may be a useful intervention to promote mTOR signalling and protein synthesis in an effort to counteract a variety of muscle wasting conditions (e.g. sarcopenia, cachexia, AIDS, inactivity/bed rest, sepsis, kidney failure, and trauma). PMID- 18403917 TI - LKB1 and AMPK and the regulation of skeletal muscle metabolism. AB - PURPOSE OF REVIEW: To address the role of LKB1 and AMP-activated protein kinase (AMPK) in glucose transport, fatty acid oxidation, and metabolic adaptations in skeletal muscle. RECENT FINDINGS: Contraction-mediated skeletal muscle glucose transport is decreased in muscle-specific LKB1 knockout mice, but not in whole body AMPKalpha2 knockout mice or AMPKalpha2 inactive transgenic mice. Chronic activation of AMPK by 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) and beta-guanadinopropionic acid enhances mitochondrial function in skeletal muscle, but AICAR or exercise-induced increases in mitochondrial markers are preserved in skeletal muscles from whole body AMPKalpha2 or muscle-specific LKB1 knockout mice. Pharmacological activation of AMPK increases glucose transport and fatty acid oxidation in skeletal muscle. Therefore, chronic activation of AMPK may be beneficial in the treatment of obesity and type 2 diabetes. SUMMARY: LKB1 and AMPK play important roles in regulating metabolism in resting and contracting skeletal muscle. PMID- 18403919 TI - Development of techniques for the analysis of isoflavones in soy foods and nutraceuticals. AB - PURPOSE OF REVIEW: For over 20 years, soy isoflavones have been investigated for their ability to prevent a wide range of cancers and cardiovascular problems, and numerous other disease states. This research is underpinned by the ability of researchers to analyse isoflavones in various forms in a range of raw materials and biological fluids. This review summarizes the techniques recently used in their analysis. RECENT FINDINGS: The speed of high performance liquid chromatography analysis has been improved, allowing analysis of more samples, and increasing the sensitivity of detection techniques allows quantification of isoflavones down to nanomoles per litre levels in biological fluids. The combination of high-performance liquid chromatography with immunoassay has allowed identification and estimation of low-level soy isoflavones. The use of soy isoflavone supplements has shown an increase in their circulating levels in plasma and urine, aiding investigation of their biological effects. The significance of the metabolite equol has spurned research into new areas, and recently the specific enantiomers have been studied. SUMMARY: High-performance liquid chromatography, capillary electrophoresis and gas chromatography are widely used with a range of detection systems. Increasingly, immunoassay is being used because of its high sensitivity and low cost. PMID- 18403920 TI - Subjective and objective nutritional assessment methods: what do they really assess? AB - PURPOSE OF REVIEW: Objective and subjective methods are used to assess nutritional status. They are used as diagnostic, prognostic and response tools. It is still controversial which of them is more suitable for each situation and what they are really assessing. The most recent findings about these methods will be discussed in this review. RECENT FINDINGS: Malnutrition still has a high prevalence all over the world. Anthropometric measurements are best useful to assess chronic malnutrition, and albumin and other visceral protein should no longer be considered as nutritional markers, but inflammatory response markers. Subjective global assessment enables comparison among different populations, and its scored version may be useful in other clinical situations besides cancer. Functional methods and bioelectrical impedance analysis may become possible to identify malnutrition in an early stage. Nitrogen balance seems to be the only way to assess the response to nutritional interventions. SUMMARY: Malnutrition should be understood as a continuum. Nutritional assessment should not be an expensive and time-consuming process, and simple methods such as subjective global assessment may be enough to identify those patients who need nutritional intervention. Future studies may show which method is more suitable to evaluate the response to this treatment. PMID- 18403918 TI - Epigenetic drugs in the treatment of skeletal muscle atrophy. AB - PURPOSE OF REVIEW: A dynamic network of anabolic and catabolic pathways regulates skeletal muscle mass in adult organisms. Muscle atrophy is the detrimental outcome of an imbalance of this network. The purpose of this review is to provide a critical evaluation of different forms of muscle atrophy from a mechanistic and therapeutic point of view. RECENT FINDINGS: The identification and molecular characterization of distinct pathways implicated in the pathogenesis of muscle atrophy have revealed potential targets for therapeutic interventions. However, an effective application of these therapies requires a better understanding of the relative contribution of these pathways to the development of muscle atrophy in distinct pathological conditions. SUMMARY: We propose that the decline in anabolic signals ('passive atrophy') and activation of catabolic pathways ('active atrophy') contribute differently to the pathogenesis of muscle atrophy associated with distinct diseases or unfavorable conditions. Interestingly, these pathways might converge on common transcriptional effectors, suggesting that an optimal intervention should be directed to targets at the chromatin level. We provide the rationale for the use of epigenetic drugs such as deacetylase inhibitors, which target multiple signaling pathways implicated in the pathogenesis of muscle atrophy. PMID- 18403921 TI - Early nutritional therapy: the role of enteral and parenteral routes. AB - PURPOSE OF REVIEW: Early nutrition is defined as the initiation of nutritional therapy within 48 h of either hospital admission or surgery. However, optimal timing for initiation of nutritional therapy through either enteral or parenteral routes remains poorly defined with the existing data. We reviewed the recent literature investigating the role of early enteral and parenteral nutrition in critical illness and perioperative care. RECENT FINDINGS: Recent studies in both trauma/surgical and nonsurgical patients support the superiority of early enteral over early parenteral nutrition. However, late commencement of enteral feeding should be avoided if the gastrointestinal tract is functional. Both prolonged hypocaloric enteral feeding and hypercaloric parenteral nutrition should be avoided, although the precise caloric target remains controversial. SUMMARY: Early enteral nutrition remains the first option for the critically ill patient. However, there seems to be increased favor for combined enteral-parenteral therapy in cases of sustained hypocaloric enteral nutrition. The key issue is when the dual regimen should be initiated. Although more study is required to determine the optimal timing to initiate a combined enteral-parenteral approach, enteral nutrition should be initiated early and parenteral nutrition added if caloric-protein targets cannot be achieved after a few days. PMID- 18403922 TI - Nutritional support in acute pancreatitis. AB - PURPOSE OF REVIEW: This review explores the role of enteral and parenteral nutrition in severe acute pancreatitis and discusses the potential benefits of glutamine, omega-3 fatty acids, arginine and selenium together with probiotics and prebiotics in these patients. In addition, the method of refeeding during the convalescent period is also examined. RECENT FINDINGS: A complex picture is emerging in which enteral nutritional support may be important early in the course of the disease with parenteral nutrition being used more as a backup and possibly only after the systemic inflammatory response has peaked. Nasogastric feeding, sometimes supplemented by parenteral nutrition, is as efficacious as nasojejunal feeding. An individualized approach, in which strategies of nutritional support are tailored to patient response, is gaining currency. Data regarding specialized formulae are mixed but the use of prebiotics is showing promise and is worthy of further exploration. In the convalescent period, preliminary data also indicate that the risk of pain developing is no greater if a light diet is instituted rather than clear fluids. SUMMARY: Nutritional support in acute pancreatitis remains challenging and controversial with a number of different and unexpected approaches, including the use of nasogastric feeding and dual enteral and parenteral nutrition support, being adopted in recent clinical trials. PMID- 18403923 TI - The impact of bariatric surgery on nonalcoholic fatty liver disease. AB - PURPOSE OF REVIEW: To analyze the effects of bariatric surgery on nonalcoholic fatty liver disease by reviewing the most important and recent studies. RECENT FINDINGS: The prevalence of obesity has increased dramatically over the last decades. Comorbidities related to obesity, such as nonalcoholic fatty liver disease are also increasing. Nonalcoholic fatty liver disease is a progressive disease with potential evolution to liver cirrhosis and hepatocellular carcinoma. Overweight patients who have nonalcoholic fatty liver disease should be considered for a weight loss program; however, long-term result with dietary interventions and drug therapy has been disappointing. Bariatric surgery is effective in promoting long-term weight loss in morbidly obese patients with control of comorbidities, especially those associated with the metabolic syndrome. On the basis of the early experience with extensive intestinal bypass, it was believed that rapid weight loss could cause liver damage. In contrast, recent prospective and retrospective observational studies and case series have demonstrated that bariatric surgery is well tolerated and is associated with nonalcoholic fatty liver disease regression in a significant number of patients. SUMMARY: There is good level of evidence that bariatric surgery is associated with nonalcoholic fatty liver disease regression in morbidly obese patients. PMID- 18403924 TI - The role of nutrition in hepatic encephalopathy. AB - PURPOSE OF REVIEW: Protein-calorie malnutrition may be observed in all clinical stages of liver disease. Nutritional management in these patients is imperative. It is crucial that protein intake is not restricted ad hoc. Administration of vegetable proteins for patients who cannot tolerate standard proteins and, if necessary, branched-chain amino acid-enriched formulae can be an option to these patients. This issue, however, remains controversial. RECENT FINDINGS: This study is an update on the nutritional management of hepatic encephalopathy based on several studies of the last decades, involving dietary protein intake and branched-chain amino acid supplementation. SUMMARY: Malnutrition is a common complication of liver disease and it adversely affects patient outcome. Inadequate dietary protein intake has a very deleterious effect on hepatic encephalopathy, nutritional status, and clinical outcome in these patients and must be avoided. The administration of branched-chain amino acids stimulates hepatic protein synthesis, reduces postinjury catabolism and therefore improves nutritional status. Conflicting results in various different trials, however, exist, and this issue remains unclear. PMID- 18403925 TI - Assessment and nutritional aspects of wound healing. AB - PURPOSE OF REVIEW: The aim of this study is to review the physiopathology and the nutritional aspects of wound healing. RECENT FINDINGS: Wound healing consists of a perfect and coordinated cascade of events that result in tissue reconstitution. The healing process is common to all wounds, independent of the agent that has caused it. It is divided didactically into three phases: inflammation, proliferation, and remodeling or maturation. Collagen is the most abundant protein in the human body and is also the main component of the wound matrix. It is organized in a thick and dynamic net, resulting from constant collagen deposition and reabsorption. Wound scar is the result of the interaction between collagen synthesis, degradation, and remodeling. There are several ways to evaluate wound healing: tensiometry, collagen morphometry, immunohistochemistry, and, more recently, the dosage of growth factors. Malnutrition adversely affects wound healing. On the contrary, the healing process can be stimulated by preoperative feeding and by certain nutrients such as glutamine, arginine, butyrate, and antioxidants. SUMMARY: Wound healing is a complex process that started to be fully understood only in recent years. Recent research has been directed to act in the nutrition modulation of the healing process. PMID- 18403926 TI - Is manganese an essential supplement for parenteral nutrition? AB - PURPOSE OF REVIEW: To summarize the role of the essential trace element, manganese, its potential toxicity, monitoring methods and dosage recommendations for nutrition support. RECENT FINDINGS: Parenteral nutrition usually contains manganese as part of a fixed concentration multiple trace element supplement. Recent literature identifies potential problems in this approach and reports toxic symptoms resulting from hypermanganesaemia in paediatric and long-term home patients. Elimination by the hepatobiliary system is frequently impaired, and parenteral administration bypasses the regulatory mechanisms of homeostasis. Together with occasional oral intake and product contamination, this can lead to brain accumulation and neurotoxicity, with individual responses to supplementation difficult to predict. Regular monitoring is recommended, but plasma and serum analyses are poor indicators of body stores. Whole blood concentrations are more accurate and correlate with signal intensity of MRI. We have identified a need for individual trace element additives to be more widely available and for multitrace element products to be reformulated. There is now a persuasive argument for not routinely adding manganese to parenteral nutrition admixtures. SUMMARY: High intravenous doses of manganese can lead to neurotoxicity. Current dosage guidelines and trace element formulations need revision. Frequent monitoring to identify tissue accumulation is recommended for paediatric and long-term home parenteral nutrition patients. PMID- 18403927 TI - Long-chain omega-3 fatty acid supply in pregnancy and lactation. AB - PURPOSE OF REVIEW: Long-chain omega-3 fatty acids are essential for the developing fetus. Docosahexaenoic acid, the most important omega-3 fatty acid, is an important component of neural and retinal membranes, and rapidly accumulates in the brain during gestation and the postnatal period. Positive associations have been shown between maternal intake of fish, seafood and omega-3 fatty acids during pregnancy and/or lactation and visual and cognitive development. RECENT FINDINGS: The review focuses on new findings by both observational and interventional studies on the influence of omega-3 fatty acids during pregnancy or lactation on gestation length and birth weight, preterm delivery, preeclampsia, maternal depression and infant visual function and neural development. SUMMARY: Omega-3 fatty acids have been associated with reduced risk of cardiovascular and other diseases. Observational and interventional studies indicate a significant association with prolonging gestation and reducing the risk of preterm delivery both in low-risk and in high-risk pregnancies. Further benefits have been suggested for intrauterine growth restriction, preeclampsia and postpartum depression, but the evidence is inconclusive. Higher maternal docosahexaenoic acid intake both in pregnancy and lactation is associated with positive infant neurodevelopmental outcomes. Women of reproductive age should achieve an average dietary docosahexaenoic acid intake of at least 200 mg/day. PMID- 18403928 TI - Leptin and regulation of linear growth. AB - PURPOSE OF REVIEW: Leptin, first identified as the product of the ob gene in leptin-deficient obese (ob/ob) mice, was originally described as a circulating hormone involved in feeding behavior and energy homeostasis. It was later found to be a pleiotropic hormone involved in the regulation of a variety of physiological processes. This review summarizes our recent understanding of the role of leptin as a linear growth-stimulating factor. RECENT FINDINGS: Leptin was found to have mitogenic effects on numerous cell types in vivo and in vitro, including several cancer cells, cells of the immune system, as well as chondrocytes of the epiphyseal growth plate. SUMMARY: Leptin stimulates linear growth by regulating the energy balance of the organism and by stimulating the production and secretion of growth hormone from the hypothalamus; at the same time, it is involved with bone remodeling and has a direct effect on the chondrocytes of the growth plate. PMID- 18403929 TI - Micronutrient needs of malnourished children. AB - PURPOSE OF REVIEW: In addition to overt manifestations of undernutrition such as stunting, wasting and underweight, micronutrient deficiencies are also recognized as important components of the spectrum of malnutrition. This review focuses on recent advances in our understanding of undernutrition and micronutrient deficiencies during childhood from a review of the literature over the last 18 months (August 2006-January 2008). RECENT FINDINGS: There is considerable advance in our understanding of the epidemiology and burden of childhood undernutrition and micronutrient deficiencies. Based on recent surveys, an estimated 32% (178 million) of children under 5 years of age were stunted. The corresponding global estimate of wasting is 10% (55 million children), of which 3.5% (19 million children) are severely wasted. It is estimated that nearly 11% of all children under 5 years of age, die due to four micronutrient deficiencies (vitamin A, zinc, iron and iodine). There is evidence from recent reviews of evidence-based interventions that administering single or multiple micronutrients can make a significant difference to health outcomes. However, delivery strategies may differ and recent data suggest that fortification may be a more efficient strategy to deliver multiple micronutrients. SUMMARY: These findings support the scaling up of evidence-based interventions to prevent and treat such deficiencies and to integrate these within health systems. PMID- 18403930 TI - Development of taste and food preferences in children. AB - PURPOSE OF REVIEW: There are concerns about the rising incidence of obesity in children and their acceptance of healthy foods. Many factors affect children's food acceptance, the most salient are those enabling early exposure to culturally appropriate foods in the weaning or pre-weaning period. Parents, however, have always observed individual differences in children's willingness to take new foods. This review looks at studies that encompass both exposure to and genetic determinants of food acceptance. RECENT FINDINGS: Children's willingness to take new foods and accept specific foods has strong-to-moderate heritability. This inherited willingness is moderated by cultural differences in early exposure to both the taste and texture of foods, giving rise to different patterns of food acceptance. Breast-feeding not only confers an advantage in food and taste acceptance but may also give rise to a preference for highly palatable 'junk food'. Modelling and flavour-conditioning may also contribute to food acceptance, whereas coaxing a child to eat may impact negatively on the intake of food. Children of obese mothers, however, react to prompting by overeating rather than food refusal. This may indicate another area where food acceptance is genetically determined. SUMMARY: Health intervention programmes that aim to promote a healthy diet in children should start at the weaning and, to a lesser extent, the pre weaning stage, and target maternal diet for optimal effectiveness. PMID- 18403931 TI - Dietary approaches to the prevention of food allergy. AB - PURPOSE OF REVIEW: The primary prevention of food allergy and atopy has become a public health priority in developed countries. Although some preventive dietary interventions have entered common practice, the level of evidence underpinning these strategies varies. RECENT FINDINGS: Exclusive breast-feeding and delayed introduction of complementary foods from the age of 4-6 months are key strategies in primary food allergy and atopy prevention. Maternal elimination diets during pregnancy and lactation are currently not recommended. Partially and extensively hydrolyzed formulae may confer a small preventive effect in high-risk infants. Amino acid-based formula is, however, considered a treatment formula and not recommended for primary allergy prevention. The effect of prebiotic oligosaccharides and probiotic bacteria in prevention of food allergy remains unclear and requires further investigation. Maternal fish oil supplementation during pregnancy shows promise in reducing the risk of allergic sensitization in the infant, but prospective long-term data are required. Fish oil supplementation in young infants was not effective. SUMMARY: Although there is evidence for the primary dietary prevention of eczema and possible respiratory allergies, data on food allergy prevention have, so far, remained less convincing. Further prospective research is needed to clarify the effectiveness of dietary interventions in primary food allergy prevention. PMID- 18403932 TI - Issues related to gluten-free diet in coeliac disease. AB - PURPOSE OF REVIEW: In the last few years, knowledge about coeliac disease has significantly improved, resulting in a better understanding of disease pathogenesis, diagnosis and therapy. This review describes the latest progress in research concerning treatment with gluten-free diet in patients with coeliac disease. RECENT FINDINGS: Gluten-free diet is generally admitted as effective therapy in symptomatic patients, but a life-long dietary treatment in some challenging cases such as 'silent' and 'latent' patients is under discussion. Tolerance to gluten may be acquired later in life, but, as latency may be transient, a strict follow-up is necessary in these patients. The composition of gluten-free diet needs a better definition; latest evidence demonstrates that oats are tolerated by most patients with coeliac disease. Finally, the amount of gluten permitted in gluten-free products is still a matter of debate; significant progress has been made in the sensitivity of techniques for gluten detection, but the daily amount of gluten that can be safely consumed is not yet defined. SUMMARY: Gluten-free diet remains the cornerstone of therapy of coeliac disease. More studies addressing the need of gluten-free diet for cases of 'potential' coeliac disease are necessary, as well as studies linking the best available analytical detection of gluten to the clinical threshold of tolerance. PMID- 18403933 TI - Bibliography. Current world literature. Anabolic and catabolic signals. PMID- 18403935 TI - Factors that affect pain during colonoscopy. PMID- 18403937 TI - Increased soluble IL-2 receptor levels during interferon and ribavirin treatment are associated with a good response in genotype 2a/2b patients with chronic hepatitis C. AB - OBJECTIVES: Serum levels of soluble interleukin-2 receptor (sIL-2R) are known to serve as a marker for the activation of T lymphocytes. We measured serum levels of sIL-2R in patients with chronic hepatitis C (CHC) during interferon (IFN) based treatment to determine the correlation between those levels and therapeutic efficacy, and to clarify whether there is a difference in the activation of T lymphocytes among HCV genotypes after the treatment. METHODS: Forty-four patients received IFN-alpha2b monotherapy (group IFN-M), whereas 82 patients received the combination therapy with IFN-alpha2b and ribavirin (group IFN+R). We measured serum sIL-2R levels in these patients before (T0) and 2 weeks (T2) after the treatment. RESULTS: The sustained virologic response rates in genotype 2a/2b patients were significantly higher than those in genotype 1b patients in both groups (P<0.005). In sustained virologic responders, sIL-2R levels at T2 were significantly higher than those at T0 in both groups (P<0.001). In nonresponders, sIL-2R levels at T2 were not different from those at T0 in group IFN-M, but were significantly higher than those at T0 in group IFN+R (P=0.0072). In genotype 1b patients, sIL-2R levels at T2 were not different from those at T0 in group IFN-M, but were significantly higher than those at T0 in group IFN+R (P=0.0064). In genotype 2a/2b patients, sIL-2R levels at T2 were significantly higher than those at T0 in both groups (P<0.0005). CONCLUSION: These findings suggest that the activation of T lymphocytes after IFN-based treatment contributes to a high sustained virologic response rate, especially in genotype 2a/2b patients. PMID- 18403936 TI - Evaluation of the French national plan to promote screening and early management of viral hepatitis C, between 1997 and 2003: a comparative cross-sectional study in Poitou-Charentes region. AB - BACKGROUND AND AIMS: The aim of this repeated cross-sectional survey was to document trends in screening practices, to analyze the evolution of the epidemiological characteristics of patients with newly diagnosed hepatitis C virus (HCV) infection, and to evaluate the implementation of hepatitis C management guidelines. METHODS: Medical laboratories in Poitou-Charentes region were surveyed on serological tests for HCV infection prescribed during two 2 month periods in 1997 and 2000, and a 4-month period in 2003. An epidemiological questionnaire and a 12-month follow-up questionnaire were addressed to physicians who prescribed tests that were positive. RESULTS: The annual screening coverage rate increased by 40% during the study period, whereas the number of positive tests fell by 53%. The estimated detection rate of new cases decreased from 43 to 26 per 100 000 inhabitants between 1997 and 2003. In 2003, 56% of serological tests were prescribed to patients who already knew that they were HCV seropositive. The frequencies of the two main risk factors (transfusion and intravenous drug use) slightly decreased. Management of newly diagnosed patients was inappropriate in 42% of cases in 1997, 33% in 2000, and 34% in 2003; 26% of the participants at the three periods declined follow-up. Among drug users, the proportion of treated patients remained stable (17%). One-third of the drug users were lost to follow-up by their family doctor. CONCLUSION: Campaigns to encourage HCV screening have been effective, but the number of newly diagnosed cases has fallen markedly. National campaigns targeting the general public and healthcare professionals seem to have had no impact on patient management: in particular, drug users still do not receive adequate follow-up. PMID- 18403938 TI - The biochemical and clinical penetrance of individuals diagnosed with genetic haemochromatosis by predictive genetic testing. AB - BACKGROUND: HFE-related genetic haemochromatosis (GH) is the commonest inherited genetic disorder in Caucasian populations with approximately one in 180 of individuals in the west of Scotland homozygous for the common C282Y mutation. The clinical diagnosis of GH, however, remains relatively uncommon - suggesting either under diagnosis or low clinical penetrance. AIM: We aimed to assess the biochemical and clinical penetrance of GH in first-degree relatives of patients with known GH, who subsequently themselves screened positive for the common GH mutations. METHODS: Individuals were identified from two large teaching hospitals in North Glasgow from July 1997 to July 2005 diagnosed with GH after predictive genetic testing after a relative was found to have GH. Details of patient history, biochemistry and known comorbidity at diagnosis and results of related further investigations were collected. RESULTS: Sixty-three individuals were identified, 31 (49%) of whom were males. Fifty-five individuals (87%) were C282Y homozygous and the remaining eight were compound heterozygotes for C282Y and H63D. All 31 male patients were found to have evidence of iron overload as opposed to 63% of females. Elevated liver enzyme levels were encountered in 15 patients (24%). All except one had evidence of iron overload. Four individuals underwent a liver biopsy, two of whom had hepatic fibrosis. Four patients were found to be diabetic. A full clinical history was obtained from 54 of 63 individuals, 38 (70%) of whom were entirely asymptomatic. Thirteen individuals complained of joint pains and a further nine complained of fatigue. CONCLUSION: This study suggests that although biochemical penetrance of GH is high, the clinical penetrance is low. PMID- 18403939 TI - Liver cirrhosis, other liver diseases, pancreatitis and subsequent cancer: record linkage study. AB - OBJECTIVE: To determine the risk of cancer in cohorts of patients with alcoholic cirrhosis, other alcoholic liver diseases, other and unspecified cirrhosis, primary biliary cirrhosis, viral hepatitis, acute pancreatitis and chronic pancreatitis compared with the risk in a control cohort. METHODS: Analysis of statistical database of linked hospital and mortality data in an area in southern England. RESULTS: Compared with the control cohort, rate ratios were elevated for cancer overall and were particularly high for liver cancer in patients with alcoholic cirrhosis (rate ratio for cancer overall 2.4, 95% confidence interval 2.0-3.0 and 27.8, 17.7-41.7 for liver cancer); with other and unspecified cirrhosis (3.1, 2.7-3.6 and 35.1, 25.4-47.6); with other alcoholic liver diseases (2.3, 2.0-2.7 and 17.7, 11.5-26.0); with primary biliary cirrhosis (1.4, 0.9-2.0 and 19.6, 8.4-39.1) and with viral hepatitis (1.5, 1.2-1.9 and 18.6, 9.8-32.2). Pancreatic cancer risk was significantly and substantially elevated in all cohorts except that with primary biliary cirrhosis. Lung cancer risk was significantly high in all cohorts except those with primary biliary cirrhosis and viral hepatitis. Oral cavity cancers were elevated in alcoholic cirrhosis cohort (8.6; 3.1-18.9) and the other alcoholic liver diseases cohort (10.1; 5.9-16.2), as was colon cancer (2.8; 1.4-5.0 and 2.0; 1.2-3.3, respectively). Non-Hodgkin's lymphoma was significantly elevated in the group of 'other and unspecified cirrhosis' (11.4; 7.2-17.3). CONCLUSION: All seven conditions carry an increased risk of cancer, but each condition has a somewhat different profile of cancer risk associated with it. PMID- 18403940 TI - Oxidative stress with altered element content and decreased ATP level of erythrocytes in hepatocellular carcinoma and colorectal liver metastases. AB - Our aim was to study the possible alterations of redox status (enzymatic and nonenzymatic parameters and metal elements) in erythrocytes of patients with hepatocellular carcinoma (HCC), colorectal liver metastases (CRLM) and benign liver neoplasms. The function of redox homeostasis is closely connected to the energy level of erythrocytes, therefore, the ATP level was also determined. Antioxidant parameters, enzyme activities of superoxide dismutase and glutathione peroxidase were estimated in the erythrocytes of 11 patients with benign tumour, 23 patients with primary malignant and 37 metastatic liver tumour patients and 30 age-matched and sex-matched healthy controls. Element content with inductively coupled plasma optical emission spectrometer and ATP level by the chemiluminometric method were also determined from the samples. Free radical intensity was significantly increased, whereas erythrocyte glutathione peroxidase and superoxide dismutase activities were significantly decreased in the HCC and CRLM groups versus benign groups and controls. Se, Mn and Zn levels were lowered in HCC and CRLM groups versus benign and control groups. The content of Cu, Mg, Se and Zn changed significantly between HCC and CRLM groups. Similarly, ATP concentration decreased in HCC and CRLM versus controls and benign groups. The lowest levels of ATP and antioxidant enzyme activities were found in the case of CRLM patients. These results reveal an alteration in the ATP level of erythrocytes with concomitant changes in the antioxidant defence system in hepatic cancer patients. Altered redox homeostasis (oxidative damage) may lead to decreased ATP level and consequently may play an important role in primary carcinogenesis and generation of metastases, as well. PMID- 18403941 TI - Risk factors associated with nonalcoholic fatty liver disease and its relationship with the hepatic histological changes. AB - OBJECTIVE: The objective of this study is to determine the association between risk factors and nonalcoholic fatty liver disease (NAFLD), and to establish the relationship between risk factors and hepatic histological changes in obese women. METHODS: A case-control design study. Women with NAFLD (cases) were compared with a control group of obese women without NAFLD matched by age, body mass index, waist circumference, and body fat. Irrespective of serum aminotransferases levels, diagnosis of NAFLD was established by the presence of type II diabetes, hypertriglyceridemia, and ultrasonographic changes of hepatic steatosis. Diagnosis of nonalcoholic steatohepatitis was performed by a liver biopsy. Women with an aspartate aminotransferase/alanine aminotransferase (ALT) ratio of at least 1 underwent liver biopsy. Alcohol consumption, hepatitis, and drugs that promote cholestasis or liver injury were the exclusion criteria. Multiple regression analysis was used to compute the association between the risk factors and NAFLD, and Spearman's analysis was used to examine its relationship with histological hepatic changes. RESULTS: A total of 108 obese women were enrolled. The frequency of high blood pressure, hypertriglyceridemia, and diabetes was similar between the groups. ALT (54.4+/-33.3 and 39.8+/-29.8, P=0.03) but not aspartate aminotransferase (45.4+/-23.1 and 36.7+/-21.2, P=0.06) was significantly higher in the women with NAFLD. The multivariate regression analysis showed a significant association of ALT (odds ratio 2.7; 95% confidence interval, 1.3-10.4), but not other variables with NAFLD. Type II diabetes was strongly correlated with ballooning and inflammation, and ALT with inflammation and fibrosis. CONCLUSION: Obese women with similar metabolic alterations exhibit different hepatic outcomes. Elevation of ALT, but not other risk factors, was associated with NAFLD. Diabetes and ALT correlate with histological hepatic changes. PMID- 18403942 TI - P-glycoprotein function and expression during obstructive cholestasis in rats. AB - OBJECTIVES: The present study was aimed at evaluation of in vivo biliary and renal excretion of rhodamine 123 (Rho123), a P-glycoprotein (P-gp) substrate, in rats during either acute or chronic cholestasis induced by bile duct obstruction (BDO). METHODS: The Rho123 clearance study was performed either one (BDO1) or seven (BDO7) days after BDO. Bile flow was reconstituted, and bile and urine were collected after steady-state plasma concentration of Rho123 was attained. Tissue expression of P-gp was evaluated by quantitative immunohistochemistry, and immunoblotting. RESULTS: Significant up-regulation of the liver P-gp protein was observed in acute and chronic cholestasis. Primary periportal location of P-gp was enlarged also to pericentral areas. In the kidneys, immunohistochemistry showed pancellular increase in P-gp after 1 day of BDO, which subsided after 7 days of BDO. Nevertheless, biliary and renal clearances (CL(Bile) and CL(R)) of Rho123 did not reflect the induction of P-gp expression. While CL(Bile) was reduced one day after cholestasis and restored on the seventh day, the CL(R) was preserved in BDO1 group and reduced in BDO7 group without change in glomerular filtration rate. In parallel, biliary and renal clearances of conjugated bilirubin were significantly reduced in both cholestatic groups compared with controls. CONCLUSION: These findings suggest that extrahepatic cholestasis causes time-dependent changes in elimination of Rho123 which do not exactly reflect alteration of P-gp expression in the rat liver and kidney. These data may help to explain impaired elimination of P-gp substrates after short-term cholestasis that may commonly occur in clinical practice. PMID- 18403943 TI - Exploring possible mechanisms for primary bile acid malabsorption: evidence for different regulation of ileal bile acid transporter transcripts in chronic diarrhoea. AB - OBJECTIVES: Chronic diarrhoea resulting from primary idiopathic bile acid malabsorption (IBAM) is common, but its aetiology is largely unknown. We investigated possible mechanisms, first looking for common sequence variations in the cytoplasmic ileal bile acid-binding protein (IBABP, gene symbol FABP6), and secondly, determining the expression of ileal mucosal transcripts for the apical sodium-linked bile acid transporter (ASBT), IBABP, the putative basolateral transporters, OSTalpha and OSTbeta, and regulatory factors. METHODS: Genomic DNA was prepared from two cohorts of patients and two control groups; the promoter and exonic regions of FABP6 were sequenced. In intestinal biopsies, transcript expression was measured by quantitative real time-PCR, using ileum from 17 patients and 21 controls. RESULTS: Sequence variations were identified in FABP6, but overall frequencies were similar in patients and controls. Transcripts of ASBT and IBABP, but not OSTalpha and OSTbeta, were expressed at higher levels in ileum than duodenum. The transcription factors farnesoid-X-receptor (FXR) and liver-receptor-homologue (LRH-1) were also more abundant in ileum, as was fibroblast growth factor 19 (FGF19), unlike short heterodimer partner (SHP), c Fos, or CDX2. No significant differences in mean or median values were found between the groups for any of these transcripts. However, findings on regression analysis suggested that these transporters differ in their regulation, particularly in the relationships of CDX2, LRH-1 and FXR with OSTalpha. CONCLUSION: Most cases of IBAM are unlikely to be caused by genetic variation in FABP6 or by major differences in transporter transcript expression. Our evidence indicates that other factors, such as regulation of expression of the basolateral bile acid transporter, should be considered as possible causes. PMID- 18403944 TI - Gluten tolerance in adult patients with celiac disease 20 years after diagnosis? AB - BACKGROUND AND OBJECTIVE: Celiac disease (CD) is believed to be a permanent intolerance to gluten. A number of patients, however, discontinue the gluten-free diet (GFD) without developing symptoms or signs. The aim of our study was to investigate whether CD patients are capable of developing tolerance to gluten. METHODS: All 77 adult patients from our hospital known to have biopsy-proven CD for more than 10 years were invited to participate. We investigated symptoms, gluten consumption, antibodies for CD and other autoimmunity, human leukocyte antigen (HLA)-typing, bone mineral density, and performed small bowel biopsies. Tolerance was defined as no immunological or histological signs of CD while consuming gluten. RESULTS: Sixty-six patients accepted participation, but after review of the diagnostic biopsies 53 were found to have true CD. Twenty-three percent of patients had a gluten-containing diet, 15% admitted gluten transgression and 62% followed the GFD. Patients on a GFD had significantly more osteoporosis. Normal small bowel mucosa was found in four of eight on gluten containing diet and in four of four with gluten transgression. Two patients were considered to have developed tolerance to gluten. One of them was HLA-DQ2/DQ8 negative. CONCLUSION: Development of tolerance to gluten seems possible in some patients with CD. Further follow-up will show whether this tolerance is permanent or only a long-term return to latency. This feature may be associated with genetic characteristics, especially with HLA genotypes that differ from DQ2 or DQ8. More insight into the mechanisms of the development of gluten tolerance may help to distinguish those CD patients that might not require life-long GFD. PMID- 18403945 TI - Does laparoscopic antireflux surgery improve quality of life in patients whose gastro-oesophageal reflux disease is well controlled with medical therapy? AB - OBJECTIVE: Both medical therapy and laparoscopic antireflux surgery have been shown to improve quality of life in gastro-oesophageal reflux disease. Although patients with poor symptom control or side effects on medical therapy might be expected to have improved quality of life after surgery, our aim was to determine, for the first time, whether patients whose symptoms are well controlled on medical therapy but who decide to undergo surgery (patient preference) would experience improved quality of life. METHODS: Retrospective analysis of our patient database (1998-2003, n=313) identified 60 patients who underwent laparoscopic antireflux surgery for the indication of patient preference. Two generic quality-of-life questionnaires (Short Form 36 and Psychological General Well-Being index) and a gastrointestinal symptom questionnaire (Gastrointestinal Symptom Rating Scale) were completed preoperatively, while on medical therapy, and 6 months after surgery. RESULTS: Thirty-eight patients completed all three questionnaires at both time intervals: 31 males, seven females; mean age 42 (15-66) years. Preoperative scores while on medical therapy were significantly improved after surgery: Short Form 36 median physical composite scores 52.0 and 54.0 (P=0.034) and mental composite scores 51.0 and 56.0 (P=0.020); Psychological General Well-Being median total scores 78.0 and 90.0 (P=0.0001); Gastrointestinal Symptom Rating Scale median total scores 2.13 and 1.73 (P=0.0007) and reflux scores 2.50 and 1.00 (P<0.0001). CONCLUSION: Laparoscopic antireflux surgery significantly improved quality of life in reflux patients whose symptoms were well controlled on medical therapy. Although on the basis of a noncomparative trial with a relatively short follow-up period, we believe such patients should be considered for laparoscopic antireflux surgery. PMID- 18403946 TI - Effects of ginger on gastric emptying and motility in healthy humans. AB - OBJECTIVE: Ginger has been reported to improve upper gastrointestinal symptoms. Little information about the effects of ginger on gastric motor function, exists, however. Our aim was to investigate the effects of ginger on gastric emptying, antral motility, proximal gastric dimensions, and postprandial symptoms. METHODS: Twenty-four healthy volunteers were studied twice in a randomized double-blind manner. After an 8 h fast, the volunteers ingested three ginger capsules (total 1200 mg) or placebo, followed after 1 h by 500 ml low-nutrient soup. Antral area, fundus area and diameter, and the frequency of antral contractions were measured using ultrasound at frequent intervals over 90 min, and the gastric half-emptying time was calculated from the change in antral area. Gastrointestinal sensations and appetite were scored using visual analog questionnaires. Data are expressed in terms of mean+/-standard error. RESULTS: Antral area decreased more rapidly (P<0.001) and the gastric half-emptying time was less after ginger than placebo ingestion (13.1+/-1.1 vs. 26.7+/-3.1 min, P<0.01), whereas the frequency of antral contractions was greater (P<0.005). Fundus dimensions did not differ, and there was no significant difference in any gastrointestinal symptoms. CONCLUSION: Ginger accelerates gastric emptying and stimulates antral contractions in healthy volunteers. These effects could potentially be beneficial in symptomatic patient groups. PMID- 18403947 TI - Helicobacter pylori infection upregulates endothelial nitric oxide synthase expression and induces angiogenesis in gastric mucosa of dyspeptic patients. AB - BACKGROUND: Helicobacter pylori (H. pylori) infection induces nitric acid (NO) overproduction through inducible NO synthase (NOS) expression, subsequent DNA damage and enhanced antiapoptosis signal transduction sequence in the human gastric mucosa, whereas its possible effect on endothelial nitric oxide synthase (eNOS) expression has not as yet been investigated. The aim of this study was to evaluate the effect of H. pylori infection in the expression of eNOS in gastric mucosa. PATIENTS AND METHODS: We prospectively studied 30 nonsmoking dyspeptic patients (12 men, 18 women, mean age 54.26+/-12.89 years). The diagnosis of H. pylori infection was based mainly on histology. The histological grading of H. pylori infection was evaluated according to the modified Sydney classification. Histological grading of eNOS expression and microvessel density as estimated by CD34 expression were determined by immunohistochemistry (degree 0-3) and correlated with H. pylori infection and histological degree of gastritis. RESULTS: Twelve patients were H. pylori-positive and 18 patients were H. pylori negative. The two groups were matched for age (P=0.139), sex (P=0.342) and similar degree of gastritis. Intensity of eNOS and CD34 expression in the corpus and antrum were significantly correlated (P<0.001). eNOS expression was correlated with H. pylori infection in the mucosa of the body and antrum (P=0.013 and 0.037, respectively) but not with gastric inflammation and activity (P=0.848 and 0.871, respectively, for the corpus and P=0.565 and 0.793, respectively, for the antrum). H. pylori-positive patients showed higher expression of CD34 positive blood vessels in the mucosa of the antrum (P=0.048). CD34 expression was correlated with gastric inflammation and activity (P=0.03 and 0.044, respectively) in the mucosa of the antrum of H. pylori-positive patients. CONCLUSION: H. pylori infection upregulates eNOS, and induces angiogenesis, contributing to H. pylori-associated pathophysiology in gastric mucosa. PMID- 18403948 TI - Proteomics in liver fibrosis is more than meets the eye. AB - Liver fibrosis is a serious health issue for many liver patients and is currently diagnosed using liver biopsy. The erroneous nature of this technique urges the search for better, noninvasive alternatives. In this regard, proteomics has been described as a useful biomarker discovery tool and has become increasingly applied in the study of liver fibrosis. Experimental and clinical studies have already provided deeper insights in the molecular pathways of liver fibrosis and even confirmed previous findings. Recent advances in proteomic strategies and tools enable multiple fractionation, multiple protein identifications and parallel analyses of multiple samples. Despite its increasing popularity, proteomics still faces certain pitfalls concerning preanalytical variability, protein coverage and statistic reliability. Proteomics is still evolving, but will undoubtedly contribute to a better understanding of the basics of the pathology and certainly offer opportunities in liver fibrosis diagnostics and therapeutics. PMID- 18403949 TI - Diminishing returns in sequential interventions of gastroenterology. AB - Gastroenterological treatment is often comprised of a series of multiple interventions, each one associated with a potential benefit, as well as harm. The analysis addresses the question of when to stop a series of consecutive interventions and maximize medical benefit. The benefit and harm associated with gastroenterological interventions are modeled as 'continuous' influences on patient health or as a 'discrete' sequence of random events. The analysis suggests that the benefit of a sequence of gastroenterological interventions is likely to accumulate following few interventions at the beginning of therapy and that any prolonged sequence is likely to inflict as much harm as benefit. It is impossible to reach a state of perfect health with inherently imperfect interventions. Health cannot be raised above a level that equals the ratio of expected benefit/(expected benefit+expected harm). As the achievement of perfect health constitutes an unattainable goal, a sequence of gastroenterological interventions should not be continued much further beyond its initial success. PMID- 18403950 TI - Lycopodium similiaplex-induced acute hepatitis: a case report. AB - A 46-year-old woman was admitted to our department with symptoms of nausea, anorexia and asthenia. Serum alanine aminotransferase and aspartate aminotransferase levels were increased; all serological tests for viral hepatitis and autoimmune disorders were negative. She had taken Lycopodium similiaplex solution as sedative for the previous 8 weeks, whose two constituents, Lycopodium serratum and Chelidonium majus, are found to be potentially toxic. After discontinuing L. similiaplex use, liver values returned to normal and she was asymptomatic. The diagnosis was definitively confirmed by liver biopsy; on the basis of the histological specimen, a hypersensitivity reaction was hypothesized as a possible pathogenic mechanism. Hepatotoxicity of phytotherapy has already been described, although so far, the true incidence and the pathogenic mechanisms are largely unknown. It is important to increase awareness of both clinicians and patients about the potential dangers of herbal remedies; surveillance systems and quality control of these products are necessary. PMID- 18403951 TI - First-time use of bevacizumab for aggressive, metastatic hepatocellular carcinoma in an HIV/hepatitis B virus coinfected patient: a case report. AB - We present a case of an HIV-1 infected patient with history of chronic hepatitis B and chronic alcohol use without cirrhosis, who presented with aggressive hepatocellular carcinoma with multiple metastases. Systemic chemotherapy combined with use of bevacizumab (anti-vascular endothelium growth factor monoclonal antibody) was without effect and the patient succumbed to his disease within few weeks. To our knowledge, this is the first report in the English literature of bevacizumab use for metastatic hepatocellular carcinoma in HIV-infected patients. PMID- 18403952 TI - Recurrence of cholangiogenous carcinoma in port-sites two years after laparoscopic removal of noncancerous gallbladder. AB - We present a unique case of carcinoma diagnosed in port-site, two years after uncomplicated laparoscopic cholecystectomy for benign cholecystitis. Analysis of morphology and cytokeratin profile (CK19+ and CK20+/-) of resected port-site tumor allows us to establish the diagnosis of tubular carcinoma with probable cholangiogenic origin. The primary carcinoma was not diagnosed in archival gallbladder tissue, despite repeated histological examination. No other primary tumor was identified during follow-up. Patient history and histological/immunohistochemical picture of the recurrent tumor suggested that primary carcinoma was probably located in the gallbladder, but was not detected during initial and repeated histological examinations of postoperative specimen. The patient is still alive, 12 months after the first port-site recurrence and 36 months after initial laparoscopy. PMID- 18403953 TI - Flow cytometric analysis of intestinal intraepithelial lymphocytes in the diagnosis of refractory celiac sprue. AB - The etiology of refractory celiac sprue (RCS) is unclear. In a high proportion of cases, the clonal nature of intestinal intraepithelial lymphocytes (IEL) can be demonstrated and a pathogenetic implication of intestinal IEL has been postulated. The prognosis of this subgroup of RCS is poor, with a high risk to develop an overt lymphoma and uncontrolled malabsorption despite steroid/immunosuppressive therapy. Cases with a relatively indolent clinical course, however, exist and their early diagnosis may be difficult. To gain insight into the pathogenic implication of intestinal IEL in refractory celiac sprue, we have performed an extensive phenotypic and functional characterization of clonal intestinal IEL in a patient with an indolent form of refractory celiac sprue, using multiparametric flow cytometry. The abnormal lymphocyte infiltrate lacked surface membrane expression of CD3/T-cell receptor (TCR) complexes (TCR( ), CD4(-), CD8(-), sCD3(-)), but contained intracellular CD3(epsilon) (CyCD3(+)) and surface CD103(+) and CD7(+). In particular, these cells showed a unique spontaneous ex-vivo cytokine secretion profile with an increased percentage of CD3(-) IEL containing TNF-alpha and IL-10, in the absence of IL-2, IL-4 and IFN gamma. Altogether our results suggest that flow cytometry immunophenotyping of intestinal IEL, in cases suspected of celiac disease and their complicated forms, could be of great help in the correct diagnosis of RCS and the understanding of the immunopathogenic mechanisms of the disease and their clinical and/or therapeutical implications. PMID- 18403954 TI - RSO interview with Mary Ellen Jafari. PMID- 18403955 TI - Dose rates from a cobalt-60 pool irradiator measured with Fricke dosimeters. AB - This project used Fricke dosimeters to determine the dose rates at multiple locations in a Co pool irradiator. Fricke dosimetry is widely accepted as a chemical dosimetry method to measure radiation absorbed dose due to its simple recipe, linear response, wide dose range, good reproducibility, ease of measurements, and low operational cost. Calibration measurements were used to determine a molar extinction coefficient of 2,185 +/- 14 L mol cm at 303 nm and 25 degrees C; the molar extinction coefficient is comparable to values from the published literature. The Fricke dosimeters measured the dose rate of a National Institute of Standards and Technology-traceable calibrated gamma radiation field to within 1.2% of the calibrated value. The pool irradiator had the largest dose rates near the middle of the torpedo, with dose rate decreasing as one moved towards the bottom or top of the torpedo. The dose rate across the torpedo is not uniform at each level, because of the non-uniform distribution of source activity around the irradiator. Relative error in the Fricke dosimeter dose rate measurements ranged from 1-2%. The dose rates mapped in this project can be used to plan bulk sample irradiation, although dosimetry measurements should still be obtained to confirm delivered dose. PMID- 18403956 TI - Guidelines for handling radioactively contaminated decedents. AB - The Centers for Disease Control and Prevention recently issued guidelines for medical examiners, coroners, and morticians in dealing with decedents after detonation of an improvised nuclear device (IND) or radiological dispersal device (RDD) (). Partners in this effort included the New York City Office of Chief Medical Examiner and the National Funeral Directors' Association. This paper describes the handling techniques required for loose surface contamination, radioactive shrapnel, and internal contamination caused by inhaling or ingesting radioactive materials from an IND or RDD, and provides suggested guidelines for medical examiners, coroners, and morticians to deal with these situations. PMID- 18403957 TI - A graded approach to flow and transport modeling to support decommissioning activities at the Savannah river site. AB - A graded approach to flow and transport modeling has been used as a cost effective solution to evaluating potential groundwater risk in support of Deactivation and Decommissioning activities at the United States Department of Energy's Savannah River Site (SRS) in Aiken, South Carolina. This approach balances modeling complexity with potential risk and has been successfully used at SRS to reduce costs and accelerate schedule without compromising human health or the environment. The approach incorporates both simple spreadsheet calculations (i.e., screening models) and complex numerical modeling to evaluate the threat to human health posed by contaminants leaching from decommissioned concrete building slabs. Simple spreadsheet calculations were used to produce generic slab concentration limits for a suite of radiological and non radiological contaminants for a chemical separations area at SRS. These limits, which are based upon the United States Environmental Protection Agency Soil Screening Guidance, were used to eliminate most building slabs from further risk assessment, thereby limiting the time and associated cost of the more rigorous assessment to higher risk facilities. Of the more than 58 facilities located in the area, to date only one slab has been found to have a contaminant concentration in excess of the area specific slab limit. For this slab, a more rigorous numerical modeling effort was undertaken which eliminated some of the simplifying and conservative assumptions inherent in the spreadsheet calculations. Results from the more sophisticated numerical model show that the remaining contaminant of concern would not likely impact groundwater above drinking water standards. PMID- 18403958 TI - Measurements should not be recorded as less than MDA: an extension. AB - Sometimes the results of measurements in radioactivity are reported as "less than the minimum detectable activity." Over the years there have been articles informing the reader that it is incorrect to express the results of measurements as less than the minimum detectable activity. A very brief review of past comments on expressing the results of measurements precedes a discussion of why measurements should not be reported as less than the minimum detectable activity. The decision level is the value of the net count above which a measurement process is claimed to have detected activity; it is determined so that the probability of detecting activity, when there is no activity in the sample, is less than or equal to the desired error of the first kind. This article extends previous work restricted to paired counting, where the blank and sample were counted for the same amount of time, to blank count times greater than or equal to the sample count times. PMID- 18403959 TI - Design and verification of shielding for the advanced spent fuel conditioning process facility. AB - An Advanced spent fuel Conditioning Process Facility (ACPF) has recently been constructed by a modification of previously unused cells. ACPF is a hot cell with two rooms located in the basement of the Irradiated Materials Experiment Facility (IMEF) at the Korea Atomic Energy Research Institute. This is for demonstrating the advanced spent fuel conditioning process being proposed in Korea, which is an electrolytic reduction process of spent oxide fuels into a metallic form. The ACPF was designed with a more than 90 cm thick high density concrete shield wall to handle 1.38 PBq (37,430 Ci) of radioactive materials with dose rates lower than 10 muSv h in the operational areas (7,000 zone) and 150 muSv h in the service areas (8,000 zone). In Monte Carlo calculations with a design basis source inventory, the results for the bounding wall showed a maximum of 3 muSv h dose rate at an exterior surface of the ACPF for gamma radiation and 0.76 muSv h for neutrons. All the bounding structures of the ACPF were investigated to check on the shielding performance of the facility to ensure the radiation safety of the facility. A test was performed with a 2.96 TBq (80 Ci) 60Co source unit and the test results were compared with the calculation results. A few failure points were discovered and carefully fixed to meet the design criteria. After fixing the problems, the failure points were rechecked and the safety of the shielding structures was confirmed. In conclusion, it was confirmed that all the investigated parts of the ACPF passed the shielding safety limits by using this program and the ACPF is ready to fulfill its tasks for the advanced spent fuel conditioning process. PMID- 18403960 TI - A multi-functional electronic program for the management of radioisotopes. AB - Everyone will agree that specialized computer programs have done away with the many tedious tasks associated with manually keeping track of radioisotopes. Enhanced electronic programs have virtually cut the time of managing radioisotopes. Agriculture and Agri-Food, Canada's (AAFC) program for the management of radioisotopes, is somewhat different from most electronic programs. It is divided into three levels of management that are dependent on the roles that a user might have when applying the application. These roles include the Departmental Radiation Safety Officer (DRSO), Radiation Safety Officer (RSO), and authorized user, which meets the requirements of the Canadian Nuclear Safety Commission. The DRSO and authorized AAFC Radiation Safety Committee members have access to the first level of management. This is the highest level of control, and only the DRSO has permission to add a nuclear substance to the system with the approval of the Canadian Nuclear Safety Commission (CNSC). This level of management is also responsible for adding authorized users, locations, and managing the Internal Use Permits. The second level of management is for site specific RSOs. They have access to all information regarding their center of activity, but they cannot change Internal Use Permit data. The RSOs can reset passwords, authorize new users, control the maximum activity limit, etc., but are limited to viewing only the information that relates to their internal use permit. However, they retain significant control within the permit. The third and last level of management is for authorized users who can access the radioisotope order-distribution-disposal section, waste or storage containers creation file, and leak/wipe test procedures. As in the case of the DRSO and RSO, they also have access to all reports and inventories for their center of activity but they cannot change Internal Use Permit or inventories data. This program has proven to be a valuable tool for scientific staff in achieving compliance and solving issues of non-compliance in the overall management of radioisotopes. PMID- 18403961 TI - Comparison of the St. Petersburg phantom with a BOMAB phantom in the ORTEC StandFast whole body counter: a Monte Carlo simulation. AB - Three sizes of the St. Petersburg phantom have been compared to six sizes of BOMAB phantoms measured by a virtual StandFast whole body counter using Monte Carlo simulations to investigate if the counting efficiencies are equivalent. This work shows that previously published data comparing the Reference Man sized phantom at 662 keV is supported; however, the simulations also show that the smaller sized St. Petersburg phantoms do not agree well with BOMAB phantoms. It is concluded that, compared with BOMAB phantoms, the St. Petersburg phantoms are system dependent and that they should be validated over a wide photon energy range against corresponding BOMAB phantoms prior to their use for calibrating whole body counters. PMID- 18403962 TI - Dose assessment for reentry or reoccupancy and recovery of urban areas contaminated by a radiological dispersal device: the need for a consensus approach. AB - Should an attack occur in an urban area with a Radiological Dispersal Device (RDD), guidance is available on the acceptable total dose equivalent for reentering the contaminated zone, and there is an accepted methodology for plume projection in the model used by the Federal Radiological Monitoring and Assessment Center (FRMAC). After initial characterization of the impact of the plume caused by an RDD, there will be considerable pressure from the public to allow them to return and quickly collect their belongings, and, eventually, to reoccupy residences and to reopen businesses. The FRMAC procedures principally deal with early and intermediate phase dose assessment, but do include late phase assessment procedures. However, the late phase assessments do not include complex geometries, such as the internal structure of buildings. This paper identifies areas where more specificity is needed to rapidly provide assessments to health officials and senior decision-makers. In this regard, there is no national consensus method to calculate projected dose inside buildings after an RDD event that addresses selecting the exposure pathways, scenarios, key parameters, etc. Therefore, to demonstrate an approach that exemplifies some of the technical and policy issues, which are unresolved, four exposure scenarios (residential, industrial, public park, park worker) were evaluated to determine the level of contamination that would deliver a dose equivalent of 10 mSv in the first year, excluding exposure during the first 4 d (emergency phase). In addition, the retrieval of personal belongings was simulated by assessing a 1-h exposure for the residential and industrial scenarios. RESRAD-BUILD was used to calculate the surface concentration of 60Co, 90Sr, 137Cs, 192Ir, 226Ra, 238Pu, 241Am, and 252Cf that would lead to a 10 mSv reference dose for these exposure periods. These example studies are intended to provide insights and guidance on how a municipal health agency can begin to develop a response plan, and to understand how the assessment process will determine the dose impacts resulting from an RDD event. The model's predictions heavily depend on the choice of the model's parameters; for several key ones, there are large uncertainties about their appropriate values. To avoid having to make hasty decisions during an emergency after an RDD attack, this analysis demonstrates that a detailed protocol for calculating dose should be developed prospectively so that decision-makers already are fully familiar with the process and its ensuing products. PMID- 18403963 TI - Reconstruction of individual radiation doses for a case-control study of thyroid cancer in French Polynesia. AB - Forty-one atmospheric nuclear weapons tests (plus five safety tests) were conducted in French Polynesia between 1966 and 1974. To evaluate the potential role of atmospheric nuclear weapons testing on a high incidence of thyroid cancer observed since 1985 in French Polynesia, a population-based case-control study was performed. The study included 602 subjects, either cases or controls, all aged less than 40 y at the end of nuclear weapons testing in 1974. Radiation doses to the thyroids of the study subjects were assessed based on the available historical results of radiation measurements. These were mainly found in the annual reports on the radiological situation in French Polynesia that had been sent to the UNSCEAR Secretariat. For each atmospheric nuclear weapons test that contributed substantially to the local deposition of radionuclides, the radiation dose to the thyroid from I intake was estimated. In addition, thyroid doses from the intake of short-lived radioiodines (132I, 133I, 135I) and 132Te, external exposure from gamma-emitted radionuclides deposited on the ground, and ingestion of long-lived Cs were reconstructed. The mean thyroid dose among the study subjects was found to be around 3 mGy while the highest dose was estimated to be around 40 mGy. Doses from short-lived iodine and tellurium isotopes ranged up to 10 mGy. Thyroid doses from external exposure ranged up to 3 mGy, while those from internal exposure due to cesium ingestion did not exceed 1 mGy. The dose estimates that have been obtained are based on a rather limited number of radiation measurements performed on a limited number of islands and are highly uncertain. A thorough compilation of the results of all radiation monitoring that was performed in French Polynesia in 1966-1974 would be likely to greatly improve the reliability and the precision of the dose estimates. PMID- 18403964 TI - Use of simulink to address key factors for radon mitigation in a Fairbanks home. AB - Hilly areas around Fairbanks, Alaska, are known to have elevated soil radon concentrations. Due to geological conditions, cold winters, and the resulting stack effect, houses in these areas are prone to higher indoor radon concentrations. Key variables with respect to radon mitigation were addressed in this paper by using a dynamic model implemented in MATLAB Simulink. These variables included the ventilation rate; the foundation flow resistance, which can be affected by sealing the foundation during the construction of a house; and the differential pressure between the subslab and the house interior, which can be affected by using a subslab depressurization system. The model was used for the scenario of a varying differential pressure and then for the scenario of a varying ventilation rate at a Fairbanks home where real-time radon concentrations were measured. The correlation coefficients between the model-predicted and measured radon concentrations were 0.96 and 0.94, for both scenarios respectively, which verified the feasibility of the model for predicting indoor radon concentrations. PMID- 18403965 TI - Development and test of a GEM-based TEPC for neutron protection dosimetry. AB - A novel tissue-equivalent proportional counter (TEPC) based on a gas electron multiplier (GEM) for measuring H*(10) for neutrons was designed and constructed. The pulse height spectra (PHS) of two different neutron sources (a 252Cf source and a AmBe source) were measured using the new TEPC. The measurements were made with the TEPC filled with two different gases (10P gas and a propane-based tissue equivalent gas) at various pressures. A computer simulation of the new TEPC, based on the Monte Carlo method, was performed to obtain the PHS for the two neutron sources. It is shown that the experimental results agree well with the simulation results for both 252Cf and AmBe neutron sources. Several outstanding problems are discussed and suggestions are made to make the GEM-based TEPC a practical neutron rem meter. The potential advantage of this novel neutron rem meter would be its low weight and compactness. PMID- 18403966 TI - The "clinic" medical-dosimetric database of Mayak production association workers: structure, characteristics and prospects of utilization. AB - To study early (deterministic) and long-term effects of radiation exposure, the "Clinic" medical-dosimetric database for the Mayak Production Association worker cohort has been established at the Southern Urals Biophysics Institute (SUBI). This paper describes the principles of organization, structure and prospects of future utilization of this database. PMID- 18403967 TI - An experimental analysis of the contribution of 224Ra and 226Ra and progeny to the gross alpha-particle activity of water samples. AB - The gross alpha-particle activity of water samples analyzed by EPA Method 900.0 is investigated as a function of residue mass and geometry, time between sample collection and analysis, and time between sample preparation and analysis for samples containing 224Ra, 212Pb, and 226Ra. It is shown that the gross alpha particle activity due to 224Ra and its progeny can be up to 10 times the 224Ra activity at collection time and that due to 212Pb progeny can be up to 3 times the 212Pb activity at collection time. In samples with roughly equal activities of 224Ra and 226Ra analyzed soon after collection, it is shown that the gross alpha-particle activity is approximately constant with time because the decay of 224Ra and its progeny is offset by the ingrowth of 226Ra progeny. PMID- 18403968 TI - Comparison of dose from radiological examination for scoliosis in children among two pediatric hospitals by Monte Carlo simulation. AB - The radiation exposures of children undergoing full spine radiography were investigated in two pediatric hospitals in Greece. Entrance surface kerma (Ka,e) was assessed by thermoluminescence dosimetry and patient's effective dose (E) was estimated by Monte Carlo simulation. All required information regarding patient age and sex, the irradiation geometry, the x-ray spectra, and other exposure parameters (tube voltage and current) were registered as well. Values of Ka,e were measured to range from 0.22 mGy to 2.12 mGy, while E was estimated to range from 0.03 mSv to 0.47 mSv. In general, all values were greater in one of the two hospitals, as higher tube currents and exposure times were used in the examinations because of the difference in radiographers' training and practice. Moreover, dose to red bone marrow was found to be between 0.01 to 0.23 mSv and dose to breast ranged between 0.02 and 1.05 mSv depending on the age, projection, and hospital. These values are comparable with literature sources. PMID- 18403969 TI - Leakage correction estimate for electret ion chamber dielectric material used for long-term environmental gamma monitoring. AB - The Idaho Department of Environmental Quality INL Oversight Program (DEQ-INL) operates an environmental gamma radiation detection network consisting of a series of high-pressure ion chambers to provide real-time ambient radiation measurements and a series of passive environmental electret ion chambers to increase coverage area and measure cumulative dose over a calendar quarter. DEQ INL has identified a consistent over-response of approximately 40% by the electret ion chambers with respect to co-located high-pressure ion chambers since 1998. DEQ-INL conducted a series of three investigations to quantify this over response. The over-response is likely attributable to a number of factors, including inherent voltage loss by the electret material not due to ionization within the chamber. One aspect of the investigation verified the manufacturer's calibration factor used to convert decrease in voltage to exposure. Additional investigations were performed that identified an average electret voltage loss of 0.2 V d(-1). When this voltage correction was applied to historical environmental data, electret ion chamber response was within 10% of the co-located high pressure ion chamber response. PMID- 18403970 TI - Comment on "the role played by stakeholders in the public debate that brought Italy out of the club of nuclear energy producers". PMID- 18403975 TI - Perioperative echocardiography. Preface. PMID- 18403976 TI - Preoperative transthoracic echocardiography: when is it useful? PMID- 18403977 TI - Basic physics of transesophageal echocardiography. PMID- 18403978 TI - Assessment of myocardial systolic function by TEE. PMID- 18403979 TI - Assessment of perioperative diastolic function and dysfunction. PMID- 18403980 TI - Assessment of valvular function and abnormalities with TEE. PMID- 18403981 TI - Assessment of intracardiac shunts. PMID- 18403982 TI - Assessment of endocarditis and intracardiac masses by TEE. PMID- 18403983 TI - Impact of TEE in noncardiac surgery. PMID- 18403984 TI - Transesophageal echocardiography in pediatric surgery. PMID- 18403986 TI - Finding evidence to support facility design decisions. AB - Nearly $200 billion of healthcare construction is expected by the year 2015, and nurse leaders must expand their knowledge and capabilities in healthcare design. The goal of this bimonthly department is to prepare the nurse leader in using the evidence-based design process to ensure that new, expanded, and renovated hospitals facilitate optimal patient outcomes, enhance the work environment for healthcare providers, and improve organizational performance. In this article, the author provides nurse leaders with examples of where to find data to guide decisions about the efficacy of specific design features in achieving desired outcomes. PMID- 18403987 TI - Coordination: new ways of harnessing complexity. AB - As systems evolve over time, their natural tendency is to become increasingly more complex. Studies in the field of complex systems have generated new perspectives on management in social organizations such as hospitals. Much of this research appears as a natural extension of the cross-disciplinary field of systems theory. This is the fourth of a 5-part series on applying complex systems science to the traditional management concepts of planning, organizing, directing, coordinating, and controlling. In this article, the concept of coordination is explored from a complex systems perspective. PMID- 18403988 TI - Collaboration: education-service affiliation agreement. AB - This biannual department, sponsored by the Council on Graduate Education for Administration in Nursing (CGEAN), provides information updates to nurse leaders on educational trends, issues, and research important for effective health system administration. CGEAN's Web site can be accessed at (http://cgean.org). PMID- 18403989 TI - Why do nurse managers stay? Building a model of engagement: part 2, cultures of engagement. AB - The short tenure of nurse managers is an urgent aspect of the leadership vacuum within the nursing shortage. The authors, who sought to build a model of nurse manager engagement in contrast to a retention model, continue to report data from a national qualitative study funded by The Robert Wood Johnson Foundation of 30 outstanding and longtime nurse managers in 6 settings. Part 1 (March 2008) described the dimensions and applications of individual engagement. In this article (part 2), the authors describe the organizational factors that contribute to engagement for nurse managers and the applications for building cultures of engagement. PMID- 18403990 TI - An evidence-based approach to nurses week celebrations. AB - It is time to examine nurses week investments. With expenses increasingly scrutinized, healthcare leaders require data-driven decisions. Managing by instinct and intuition is both inadequate and reckless. This survey of 727 registered nurses identifies celebratory options for nurses week that nurses find meaningful. Knowing what registered nurses value will guide approaches to an effective nurses week activity planning. PMID- 18403991 TI - Developing nursing leaders: an overview of trends and programs. AB - Nurses need to acquire new leadership skills to meet the demands of their positions. Until recently, the healthcare industry has focused on continuing education to meet licensure requirements and to achieve advanced clinical certifications. Given increasingly complex leadership challenges, we are seeing the emergence of leadership development programs designed to enhance leadership effectiveness in the service of broader strategic objectives. By learning more about these opportunities, nurse executives can make better decisions for themselves and other nursing colleagues. PMID- 18403992 TI - Recruiting, retaining, and advancing careers for employees from underrepresented groups. AB - In response to the need to increase the number of persons from underrepresented groups working in acute care settings, 2 hospitals in upstate New York implemented a multidimensional workforce development project targeting existing and new employees meeting federally defined poverty levels. The authors describe the project and its outcomes, which surpassed original expectations. PMID- 18403993 TI - Transforming care at the bedside: an ambulatory model for improving the patient experience. AB - Transforming Care at the Bedside is a nationwide effort to design a model for improving care to hospitalized patients. With the projected growth of ambulatory services, it is increasingly important to focus on potential methods to increase patient satisfaction and care delivery improvement in the outpatient setting, as well. The authors describe the University of Pittsburgh Medical Center Hillman Cancer Center's adaptation of the Transforming Care at the Bedside care delivery improvement model to its ambulatory services arena and its promising results. PMID- 18403994 TI - Magnet recognition program: the next generation. AB - Twenty-five years ago, the foundation of the Magnet Recognition Program was established. Magnet designation has served as the hallmark of excellence for nursing practice, and research has demonstrated a profound impact on nursing practice and patient care. The purpose of this article was to forecast the direction of the Magnet Recognition Program. The authors discussed the results of a multivariate structural analysis of the forces of magnet and the subsequent future model for Magnet. PMID- 18403995 TI - The role of cardiovascular magnetic resonance in the diagnosis and management of cardiomyopathies. AB - With the improvement of noninvasive imaging, the cardiology community has to learn how to exploit properly the novel diagnostic tools in order to pursue an accurate diagnosis and better patient management. In the past decade, cardiovascular magnetic resonance has been recognized as a valuable means of studying and characterizing cardiac disorders. Notably, with the improvement of functional and contrast-enhanced imaging, cardiovascular magnetic resonance has achieved excellence in the depiction of cardiac abnormalities, providing clinicians and researchers with a unique, comprehensive approach. This review focuses on the added value of cardiovascular magnetic resonance over other conventional techniques in the diagnosis and management of the most common cardiomyopathies. PMID- 18403996 TI - The role of adiposity as a determinant of an inflammatory milieu. AB - With the growing prevalence of obesity, scientific interest in the biology of adipose tissue has been extended to the secretory products of adipocytes, since they have been shown increasingly to affect several aspects of the pathogenesis of obesity-related diseases. Until relatively recently, the role of adipose tissue itself in the development of obesity and its consequences was considered to be a passive one. It is now clear that, in addition to storing energy in the form of triglycerides, adipocytes also secrete a large variety of proteins, including cytokines, chemokines and hormone-like factors. This production of proatherogenic chemokines by adipose tissue is of particular interest, since their local secretion, for example by perivascular adipose depots, may provide a novel mechanistic link between obesity and associated vascular complications. PMID- 18403997 TI - Mechanical dyssynchrony and functional mitral regurgitation: pathophysiology and clinical implications. AB - Functional mitral regurgitation (FMR) is a common finding in patients with ischemic or nonischemic dilated cardiomyopathy as a complication of left ventricular (LV) dysfunction and remodeling associated with a fibrotic remodeling response of mitral leaflets to abnormal valvular loading. Although mitral valve tenting is the main determinant of FMR, clinical and experimental observations suggest that intraventricular delay could be a potential co-determinant of FMR. LV dyssynchrony can potentially contribute to FMR by several mechanisms, such as creating an uncoordinated regional LV mechanical activation in segments supporting the papillary muscles, determining diastolic mitral regurgitation, reducing the sphincteric function of the mitral annulus, and decreasing the efficiency of LV contraction and closing forces. Cardiac resynchronization therapy has been demonstrated to reduce FMR with correction of some of the underlying pathophysiological mechanisms. The present review article focuses on the role of mechanical dyssynchrony as a pathophysiological determinant of FMR, and on the potential role of cardiac resynchronization therapy as a therapeutic option for treatment of FMR in patients with severe heart failure and advanced LV dysfunction. PMID- 18403998 TI - Survival and hospitalization in a nurse-led domiciliary intervention for elderly heart failure patients. AB - AIM: Heart failure in the elderly population represents a complex clinical situation associated with frequent hospitalizations and numerous comorbidities. The present study aimed to evaluate the impact of a domiciliary-based nurse-led strategy in a group of very elderly patients affected by heart failure who were regularly seen at an outpatient heart failure clinic (HFC). METHODS: Patients were periodically assessed in their homes by two trained nurses under supervision of the cardiologists of the HFC. During each visit, the nurses examined clinical status and adherence to medication. When necessary, they also gathered venous blood samples for laboratory analysis and recorded an electrocardiogram. In addition, they provided key information regarding disease management to patients as well as to their care givers. During the baseline visit and, subsequently, every 6 months, an echocardiogram was performed at the HFC. RESULTS: Forty-four patients (52.4% male, median age 82 years) were followed up for a mean of 25 +/- 12 months. Compared to an equally long time period before randomization, during follow-up, a significant reduction in cardiac hospitalizations (from 1.83 +/- 1.54 to 1.07 +/- 1.39, P = 0.004), total hospitalizations (from 2.09 +/- 1.71 to 1.52 +/- 1.68, P = 0.003), HFC visits (from 3.31 +/- 2.33 to 2.24 +/- 1.38, P = 0.03) and New York Heart Association (NYHA) class (from 2.74 +/- 0.70 to 2.49 +/- 0.61, P = 0.04) was observed. Total 1-year mortality was 25% and was predicted by several clinical (weight loss, NYHA class), laboratory (hyperuricaemia, anaemia, renal failure, hyposodiemia) and echocardiographic (end-systolic diameter, ejection fraction, systolic pulmonary artery pressure) parameters. Multivariate analysis revealed that hyperuricaemia was as an independent predictor of mortality (odds ratio = 1.53, P = 0.038). CONCLUSIONS: The present study demonstrates that a domiciliary-based strategy in elderly patients affected by heart failure guarantees clinical stability and reduces hospitalizations as well as outpatient visits. PMID- 18403999 TI - How many patients would be misclassified using M-mode and two-dimensional estimates of left atrial size instead of left atrial volume? A three-dimensional echocardiographic study. AB - OBJECTIVES: Left atrial size has shown prognostic importance in a variety of cardiac conditions. Diameters, area, and volume derived from M-mode and two dimensional (2D) echocardiography are commonly used to estimate left atrial size. However, M-mode and 2D measures of left atrial size rely on various geometrical assumptions and their accuracy remains to be determined. To address this issue, we compared M-mode and 2D parameters routinely used to estimate left atrial size with three-dimensional (3D) echo measured left atrial volume (LAV) as a reference standard. METHODS: We studied 104 patients (55% males, 62 +/- 15 years, range 10 87 years), presenting for a routine echocardiographic evaluation. RESULTS: The mean 3D LAV for the study population was 90 +/- 68 ml (range 24-458 ml). We found highly significant (P < 0.0001) correlations between 3D LAV and left atrial anterior-posterior (r = 0.78, 95% CI = 0.69-0.85), superior-inferior (r = 0.74, 95% CI = 0.63-0.81) and medial-lateral (r = 0.91, 95% CI = 0.86-0.93) diameters. A highly significant correlation was also found between 3D LAV and left atrial area (r = 0.94, 95% CI = 0.91-0.96). However, using M-mode anterior-posterior diameter or left atrial area would have misclassified 57% and 70% of our study patients, respectively, regarding the degree of left atrial dilatation. Closer correlations and narrower confidence intervals were found between 3D LAV and single-plane (r = 0.98; 95% CI = 0.94-0.97) and biplane (r = 0.97; 95% CI = 0.96 0.98) 2D LAVs. CONCLUSION: Left atrial diameters and area measurements were poor predictors of 3D LAV, especially in the enlarged left atria. Therefore, these parameters can be misleading in assessing the severity of left atrial dilatation. Two-dimensional LAVs are accurate in estimating 3D LAV. The small additional accuracy obtained by using the biplane instead of the single-plane area-length method, and the fact that the biplane method is more technically demanding and time consuming, may allow the use of the area-length for routine clinical use. PMID- 18404000 TI - Usefulness of multislice computed tomography to assess patency of coronary artery stents versus conventional coronary angiography. AB - BACKGROUND: The aim of the present study was to assess the in-stent restenosis and occlusion of coronary artery stents by multislice computed tomography (MSCT) compared with conventional coronary angiography in patients with atypical chest pain and not practicable/non-conclusive stress test. METHODS: Between December 2004 and March 2006, 81 patients were scheduled and of these 72 (65 men, mean age 61 years) with 90 stents underwent MSCT angiography using a 16-slice scanner, Toshiba Aquilion 16, 8-12 months after stent placement. RESULTS: Of the 90 stents, 71 (79%) could be assessed and 19 (21%) were excluded because the image quality at the stent level was incompatible with diagnostic assessment. This results in sensitivity, specificity, and positive and negative predictive values for all assessable stents in the identification of occlusion and/or in-stent restenosis of 82, 96, 87, and 94%, respectively. When the 19 uninterpretable stents were included in the analysis, the diagnostic accuracy of MSCT in detecting in-stent restenosis and occlusion resulted in a sensitivity of 82%, specificity of 71%, positive predictive value of 40%, and negative predictive value of 94%. CONCLUSION: The results of the study suggest that MSCT angiography is a useful method for evaluating patency/occlusion of large (>or=3 mm) coronary stents in symptomatic patients with atypical chest pain and concomitant not practicable/non-conclusive exercise or stress imaging test. PMID- 18404001 TI - Comparison of standard and bicaval approach in orthotopic heart transplantation: 10-year follow-up. AB - BACKGROUND: The present study compared 86 patients who underwent orthotopic heart transplantation by bicaval and standard techniques. METHODS: Patients already followed in the first year after heart transplantation were studied at 10 years of follow-up, this time evaluating the prevalence of arrhythmias, conduction disturbances and mitral or tricuspid regurgitation. RESULTS: The following arrhythmias were observed: atrial fibrillation (one patient from the standard group), ventricular premature beats and ventricular fibrillation (each in one patient from the bicaval group). Conduction disturbances were found in 25 patients in the standard group (48.1%) and in 19 patients in the bicaval group (55.9%, P = 0.515). Twenty patients had a mono- or bifascicular block in the standard group (38.5%) versus 19 patients in the bicaval group (55.9%); furthermore, five patients in the standard group (9.6%) and none in the bicaval group had a permanent pacemaker (Fisher's exact test: P = 0.074). Mitral regurgitation was present in 13 (26.5%) and five (16.1%) patients, respectively, in the standard and bicaval groups (Fisher's exact test: P = 0.411): it was grade 1 in 12 and five patients and grade 2 in one and zero patients, respectively. Tricuspidal regurgitation was observed in 26 (53.1%) and 13 (41.9%) patients, respectively, in the standard and bicaval groups (Fisher's exact test: P = 0.366): it was grade 1 in 23 and 13 patients and grade 2 in three and zero patients, respectively. Cumulative survival was 75% at 10 years from transplant in this relatively old population of patients (mean age = 58 years). CONCLUSION: In conclusion, our data do not support any definite mandate for either of the surgical techniques. PMID- 18404002 TI - Ebstein's anomaly: a potential cause of misinterpretation of myocardial perfusion scintigraphy. AB - We present the case of a 43-year-old woman with a history of Ebstein's anomaly, who was referred for a myocardial perfusion single-photon emission computed tomography (SPECT) study due to angina-like symptoms. Dilatation of the right ventricle and right atrium, apical displacement of the septal tricuspid leaflet, small left ventricle with good left ventricular and a moderate degree of tricuspid regurgitation with mild pulmonary hypertension were found on echocardiography. Myocardial SPECT perfusion imaging with [99m Tc]tetrofosmin demonstrated a large defect in the inferior septal wall in the stress study with mild partial reversibility at rest. Coronary angiography revealed normal coronary arteries. Histological studies have shown that these patients develop increased fibrosis in the left ventricular wall and ventricular septum, especially in the basal and middle region of the septum, which adjoins with the atrialized component of the right ventricle. This is in concordance with the findings of the myocardial perfusion scan in the case described. PMID- 18404003 TI - Near-drowning syndrome: a possible trigger of tako-tsubo cardiomyopathy. AB - We report a case of transient tako-tsubo cardiomyopathy characterized by an unusual trigger in a woman victim of near-drowning syndrome. After 24 h, electrocardiogram changes and a typical echocardiographic pattern of apical ballooning with a mild increase of serum troponin level induced the suspicion of tako-tsubo cardiomyopathy despite the absence of chest pain. Left ventriculography confirmed the apical ballooning, and coronary angiography revealed normal coronary arteries. Electrocardiogram changes and apical contraction abnormalities were reversed within 1 month. In conclusion, we hypothesize that hypoxemia related to near-drowning syndrome could have induced transient myocardial dysfunction mediated by a sympathetic nerve activation. PMID- 18404004 TI - Double-outlet right ventricle in a neonate with Cantrell's syndrome. AB - We report a case of the association of Cantrell's syndrome with double-outlet right ventricle in a neonate. PMID- 18404005 TI - Infective endocarditis in the real world: the Italian Registry of Infective Endocarditis (Registro Italiano Endocardite Infettiva - RIEI). AB - In the last 30 years, major improvements have been made in understanding the pathogenesis, diagnosis and treatment of infective endocarditis. Nevertheless, mortality still remains high, close to 30-40% at 1 year, and its reduction remains the main challenge. Moreover, important epidemiological changes have been recorded. Social changes in Western countries have led to an increase in the mean length of life, and thus in degenerative valvular diseases, whereas rheumatic heart disease has almost disappeared. Increasing medicalization has led to a rise in complications and diseases related to longer hospital stay, surgical therapies and other invasive interventions. At the same time, there is an increase in immunosuppressive therapies, diseases such as cancer, diabetes mellitus and renal insufficiency that may enhance the disease. Further knowledge is needed for specific subgroups to improve both treatment and prognosis. Nevertheless, randomized trials are lacking to guide the management of the disease, and the role and indications of antibiotic prophylaxis are still the subject of debate. International multicenter studies are providing new important findings based on the experience of tertiary centers; these results may reflect referral biases. The proposal of an Italian national registry on infective endocarditis (RIEI) will overcome these limitations and provide a wide picture of the national presentation of the disease. The aim of the registry is to improve the management of infective endocarditis, through a better understanding of demographic, clinical, therapeutic and prognostic features of the disease in the real world. The background, rationale, aims and expected results of the registry are reviewed. PMID- 18404007 TI - A rare mark of anomalies of cardiac venous drainage: unusual course of pacemaker lead. PMID- 18404006 TI - Guideline recommendations for heart complications in thalassemia major. AB - Thalassemia major is an inherited hemoglobin disorder resulting in a chronic hemolytic anemia. Transfusion therapy together with elevated gastrointestinal absorption of iron determines iron overload, which causes most of the mortality and morbidity associated with the disease. Heart complications represent the leading cause of mortality in this disease, although, because of an improvement in chelation treatment, an important and progressive increase of life expectancy mainly as a result of a reduction in mortality due to cardiac dysfunction has been demonstrated in recent years. Clinical pictures of heart damage range from the involvement of the ventricles to pulmonary hypertension or symptomatic ventricular or supra-ventricular arrhythmias. For this reason, the possibility of having specific recommendations is noteworthy. These recommendations outline the definition, the follow-up and the treatment of the main heart complications in this group of patients. The identification of topics and the nomination of the committee were made on behalf of the Society for the Study of Thalassemia and Hemoglobinopathies (SoSTE). The document obtained the auspices of ANMCO, SIC, SIRM and the Cardiovascular Magnetic Resonance Working Groups of the ANMCO, SIC and SIRM. All recommendations provided in this document have been performed according to the American Cardiology College (ACC) and American Heart Association (AHA) guidelines. Moreover, the recommendations were reviewed by two external referees before the definitive approval. PMID- 18404008 TI - Exercise prescription for the prevention and treatment of cardiovascular diseases: part I. AB - Epidemiological, clinical and laboratory studies have provided definitive evidence that physical activity is able to improve fitness and reduce cardiovascular morbidity and mortality. Moreover, physical exercise also seems to significantly reduce the risk of developing other chronic diseases such as obesity, osteoporosis, diabetes, tumours and depression. Promoting physical activity in the general population is therefore one of the primary objectives of our healthcare institutions. Although the benefits of an active lifestyle have been demonstrated by numerous scientific data, only a few numbers of Italians and Europeans take up regular physical exercise. To promote physical activity, both in the general population and in subjects affected by cardiovascular diseases, the Italian Federation of Sports Medicine, the Italian Society of Sports Cardiology, the Italian Association of Hospital Cardiologists, the Italian Society of Cardiology, the Italian Association of Out-of-Hospital Cardiologists and the Italian Group of Cardiac Rehabilitation have promoted the constitution of a Task Force made up of experts in the fields of sports cardiology. The document produced by the Task Force is intended for healthcare professionals and deals with the role of physical activity in the prevention and treatment of cardiovascular diseases. It examines the beneficial effects of physical activity on the cardiovascular system, while analysing the possible risks involved and how they can be avoided. The rational principles underlying the prescription of physical activity in the cardiologic setting are described, as are the modalities for prescribing such activity. PMID- 18404009 TI - Introduction: the ACOEM depression in the workplace project. PMID- 18404011 TI - Etiology of depression and implications on work environment. AB - OBJECTIVE: We describe the etiology of clinical forms of depression and their interaction with workplace factors. METHODS: We performed selective literature searches in pertinent research areas and reviewed the literature in the context of clinical experience in several depression programs at Johns Hopkins. RESULTS: Substantial research exists about the causes of depression. The causes of clinical forms of depression are complex. Genetic factors, several specific brain disorders, gender, as well as life events or circumstances all are involved in the causal pathway. Research addressing how the work environment impacts depression and its role in aggravating or alleviating depression is more limited. CONCLUSIONS: Research findings about the various causal connections in depressive disorders may provide some guides to treatment as well as to further research on depression in the workplace. PMID- 18404012 TI - Diagnosis and referral of workplace depression. AB - OBJECTIVE: Effective treatment requires understanding of the many possible reasons for employees and patients to complain of "depression." METHODS: This process of differential diagnosis includes panic anxiety, thyroid and other medical conditions, as well as several distinct types of depression (including atypical depression and melancholia). RESULTS: Much of workplace depression care can be delivered by occupational health and mental health professionals. Optimal treatment requires accurate and specific diagnosis, and focused care. And, some cases require urgent psychiatric referral, while less urgent referral is important for some others. CONCLUSIONS: Optimal diagnosis and specific treatment is a cost effective approach that saves money for employers, while helping employees. PMID- 18404010 TI - The prevalence and correlates of workplace depression in the national comorbidity survey replication. AB - OBJECTIVE: To review evidence on the workplace prevalence and correlates of major depressive episodes, with a particular focus on the National Comorbidity Survey Replication, the most recent national survey to focus on these issues. METHOD: Nationally representative survey of Diagnostic and Statistical Manual, 4th Revision Mental Disorders. RESULTS: A total of 6.4% of employed National Comorbidity Survey Replication respondents had 12-month major depressive disorder. An additional 1.1% had major depressive episodes due to bipolar disorder or mania-hypomania. Only about half of depressed workers received treatment. Fewer than half of treated workers received care consistent with published treatment guidelines. CONCLUSIONS: Depression disease management programs can have a positive return-on-investment from the employer perspective, but only when they are based on best practices. Given the generally low depression treatment quality documented here, treatment quality guarantees are needed before expanding workplace depression screening, outreach, and treatment programs. PMID- 18404013 TI - What does research tell us about depression, job performance, and work productivity? AB - OBJECTIVE: To assess the work impact of depression. METHODS: A review of research articles published since 2002, reporting on the magnitude and/or nature of depression's impact on work. RESULTS: This research is characterized by the use of three outcome indicators (employment status, absenteeism, and presenteeism metrics) and three research designs (population-based, workplace, and clinical). The literature documents that, compared to non-depressed individuals, those with depression have more unemployment, absences, and at-work performance deficits. Methodological variation makes it difficult to determine the magnitude of these differences. Additionally, the research suggests that the work impact of depression is related to symptom severity and that symptom relief only partly reduces the adverse work outcomes of depression. CONCLUSIONS: Research has contributed to knowledge of the multidimensional work impact of depression. Further developing intervention research is an important next step. PMID- 18404014 TI - Challenges and opportunities for preventing depression in the workplace: a review of the evidence supporting workplace factors and interventions. AB - OBJECTIVE: To explore the literature regarding prevention of depression in the workplace. METHOD: Literature review of what the author believes are seminal articles highlighting workplace factors and interventions in preventing depression in the workplace. RESULTS: Employees can help prevent depression by building protective factors such as better coping and stress management skills. Employees may be candidates for depression screening if they have certain risk factors such as performance concerns. Organizational interventions such as improving mental health literacy and focusing on work-life balance may help prevent depression in the workplace but deserve further study. CONCLUSION: A strategy to prevent depression in the workplace can include developing individual resilience, screening high-risk individuals and reducing that risk, improving organizational literacy, and integrating workplace and health care systems to allow access to proactive quality interventions. PMID- 18404015 TI - A brief review of antidepressant efficacy, effectiveness, indications, and usage for major depressive disorder. AB - OBJECTIVE: Antidepressants treat major depressive disorder (MDD) with the burden of associated side effects and difficulties with compliance. The purpose of this article is to review the efficacy and effectiveness of antidepressants for MDD. METHODS: The authors conducted a focused review of selected key issues and references relevant to the clinically relevant pharmacologic treatment of MDD. Principles of treatment are reviewed. Antidepressants reviewed include SSRIs, mixed norepinephrine or serotonin uptake inhibitors, dopamine or norepinephrine uptake inhibitors, norepinephrine uptake inhibitors, antidepressants with mixed properties, and monoamine oxidase inhibitors. Augmentation and psychotherapy strategies are reviewed. RESULTS: Antidepressant efficacy has been established in randomized clinical trials and effectiveness studies for acute and long-term treatment, but many patients do not achieve remission. Augmentation strategies and focused psychotherapy can be helpful. CONCLUSIONS: Antidepressants help most patients with MDD but some are resistant to treatment and have a difficult long term course. PMID- 18404016 TI - Evidence-based psychotherapies for depression. AB - OBJECTIVE: This article briefly reviews the advantages and limitations of empirically validated psychotherapies for depression, which constitute important alternatives to somatic antidepressant treatments. METHODS: Medical literature review. RESULTS: The author describes the theories and techniques underlying cognitive behavioral therapy (CBT), interpersonal psychotherapy (IPT), behavioral therapy, and other therapies, as well as their use in individual, group, and alternative formats. CONCLUSION: The author argues that psychotherapies have been underused as antidepressant treatments relative to pharmacotherapies. PMID- 18404017 TI - Assessment of occupational impairment and disability from depression. AB - OBJECTIVE: Examination of the relationship of impairment to disability in the work psychiatry context and identification of practical strategies for occupational physicians to apply to screening, management, and appropriate referral. METHODS: Medical literature review. RESULTS: The determination and differentiation of impairment and disability is a complex psychiatric task which requires consideration of the type of employment, assessment of depression related functional impairments that can create disability for a particular occupation, and individual factors. CONCLUSIONS: The authors propose a new and more consistent strategy for identifying impairment severity and its impact on employment, including simple procedures to screen for depression and guidelines to minimize role and boundary confusion. PMID- 18404018 TI - Gaps in depression care: why primary care physicians should hone their depression screening, diagnosis, and management skills. AB - OBJECTIVE: To highlight the potential role that occupational and environmental physicians can play in providing high quality depression care by describing: the profound impact that depression can have on development and persistence of unexplained medical symptoms such as pain; the impact of depression on social and occupational functioning; the adverse effect of comorbid depression in patients with chronic medical illness on self-care and medical outcomes; and the significant direct and indirect costs associated with depression. METHOD: Literature review. RESULTS: Despite evidence-based treatment guidelines, suboptimal depression care persists. CONCLUSIONS: Building on the tenets of the chronic illness care model as well as the principles of public health, three approaches are required to address the gaps between knowledge and practice: 1) improve accuracy of diagnosis; 2) prevent chronic depression; and 3) prevent relapse and recurrence. PMID- 18404019 TI - Population-based care of depression: team care approaches to improving outcomes. AB - OBJECTIVE: To describe evidence-based quality improvement interventions in the primary care system that have been shown in randomized trials to the improve quality of care and outcomes of patients with depression. METHODS: Medical literature review, focused on the concept of population-based care and research proven ways to decrease the prevalence of depression in primary care, including several meta-analyses that described the effect of collaborative care interventions in improving the quality and outcomes of primary care patients with depression. RESULTS: A total of 37 randomized trials of collaborative care interventions have shown that collaborative care, compared with usual primary care, is associated with 2-fold increases in antidepressant adherence, improvements in depressive outcomes that last up to 2 to 5 years, increased patient satisfaction with depression care, and improved primary care satisfaction with treating depression. From a health plan perspective, cost-effectiveness analyses suggest that for most depressed primary care patients, collaborative care is associated with a modest increase in medical costs, but markedly improved depression and functional outcomes. The few studies that have used a societal perspective that included examination of both direct and indirect costs found that collaborative care was associated with overall cost savings. For patients with depression and diabetes and depression and panic disorder, there is evidence that the increase in mental health care costs associated with collaborative care is offset by greater savings in medical costs. CONCLUSION: Collaborative care is a high value intervention associated with improved quality of care, depression outcomes, and improved patient and primary care physician satisfaction. PMID- 18404020 TI - Making the business case for enhanced depression care: the National Institute of Mental Health-harvard Work Outcomes Research and Cost-effectiveness Study. AB - OBJECTIVE: Explore the business case for enhanced depression care and establish a return on investment rationale for increased organizational involvement by employer-purchasers. METHOD: Literature review, focused on the National Institute of Mental Health-sponsored Work Outcomes Research and Cost-effectiveness Study. RESULTS: This randomized controlled trial compared telephone outreach, care management, and optional psychotherapy to usual care among depressed workers in large national corporations. By 12 months, the intervention significantly improved depression outcomes, work retention, and hours worked among the employed. CONCLUSION: Results of the Work Outcomes Research and Cost effectiveness Study trial and other studies suggest that enhanced depression care programs represent a human capital investment opportunity for employers. PMID- 18404021 TI - Depression in the workplace: the role of the corporate medical director. AB - Depressive disorders are a major health issue in the US workplace. They are responsible for significant direct and indirect costs to the employer in terms of medical and pharmaceutical costs, time absent from work and decreased productivity while on the job (presenteeism). The Corporate Medical Director (CMD) or occupational health physician must be equipped to respond to this health problem just as they must be able to respond to the more "traditional" workplace issues such as communicable disease, occupational exposures, and work-related ergonomic injuries. An integrated response by the CMD includes forming partnerships with relevant departments such as the Employee Assistance Program, Human Resources, Corporate Benefits, and others; measuring the impact of the disease; and providing leadership with regard to interventions in health plan design, disability management, workplace policy, and education aimed at increasing awareness and destigmatization. PMID- 18404022 TI - Integrated management of depression: improving system quality and creating effective interfaces. AB - Depression is a chronic recurrent condition and is a leading cause of work disability. Improving occupational outcomes for depression will require an integrated approach that incorporates best practices from the clinical, community, and workplace systems. This article briefly reviews recent quality improvement initiatives and promising practices in each system and then shifts to the importance of systems integration. An integrated chronic care model uses a sophisticated case management process to support essential relationships, facilitate key plans, and efficiently link the three systems to optimize clinical, economic, and occupational outcomes. An expanded role for employers and their agents in the management of depression and other chronic diseases is seen as fundamental to maintaining a healthy and productive workforce. OBJECTIVE: To improve occupational outcomes for depression by integrating best practices from the clinical, community, and workplace systems. METHODS: After a brief review of quality improvement initiatives and promising practices in each system, an integrated chronic care model is introduced. RESULTS: A case management process that links critical systems, supports essential relationships, and facilitates key plans is expected to result in improvements in clinical, economic, and occupational outcomes. CONCLUSIONS: Employers should be more engaged with clinical and community partners in the prevention and control of depression in affected employees. PMID- 18404023 TI - Research on depression in the workplace: where do we go from here? AB - OBJECTIVE: This article overviews the current status of research on depression in the workplace and makes recommendations for future research to increase knowledge and close the research to practice gap. METHOD: Medical literature review. RESULTS: Knowledge gaps are identified along the public health continuum from prevention and promotion to disability management and return-to-work. The role of Occupational Physicians in generating and disseminating knowledge is discussed. CONCLUSIONS: The quality of research on depression in the workplace is improving rapidly but there is a need for a formal research agenda to identify priorities for a systematic multi-stakeholder effort to close critical knowledge gaps and more effectively disseminate and implement research findings in the workplace. PMID- 18404024 TI - A clinical perspective on workplace depression: current and future directions. AB - OBJECTIVE: To identify and propose potential solutions to challenges encountered by clinical and consulting occupational and environmental medicine practitioners in the management of workplace depression. METHOD: A review of English-language medical literature from 1960 to the present. RESULTS: Numerous consequences and challenges to clinical management of workplace depression are recognized. Several potential present and future roles of the practitioner--in prevention, initial recognition and management, appropriate referral, care integration, stakeholder advocacy and education, and administration--are identified, and specific interventions are proposed and explored. CONCLUSION: Numerous opportunities exist for clinical and consulting practitioners to intervene to improve the current management of workplace depression. PMID- 18404027 TI - The economics of trauma. PMID- 18404028 TI - The effect of compensation on general health in patients sustaining fractures in motor vehicle trauma. AB - OBJECTIVES: The receipt or pursuit of compensation after injury has been associated with poor outcomes. This study aims to determine the association between compensation-related factors and general health in patients with fractures sustained in motor vehicle trauma. DESIGN: Prospective survey. SETTING: Metropolitan trauma centers. PATIENTS/PARTICIPANTS: The study population was patients aged 18 years and older, presenting acutely with at least one fracture involving the long bones, pelvis, patella, talus, or calcaneus, resulting from motor vehicle trauma, and presenting acutely to 1 of 15 hospitals. INTERVENTION: Patients were surveyed on admission to determine general factors, injury factors, and socioeconomic factors. Employment status at follow-up, compensation-related factors, and the main outcome variables were measured by survey at 6 months after injury. Multiple regression was used to determine significant predictors of outcome. MAIN OUTCOME MEASUREMENT: Physical and mental health summaries of the SF 36 General Health Survey. RESULTS: Of the 306 patients recruited to the study, five were excluded, and completed questionnaires were available for 232 (75.8%). Claiming compensation was strongly associated with poor physical and mental health on univariate analysis, but it was not significant on multivariate analysis. The use of a lawyer in relation to the injury was the most significant variable associated with poor physical and mental health, after adjusting for other factors. CONCLUSIONS: Lawyer involvement, rather than pursuit of compensation, is associated with poor general health after fractures sustained in motor vehicle injuries. Although this may represent a direct effect, further research is recommended to determine the cause for this association. PMID- 18404029 TI - Orthopaedic traumatology: the hospital side of the ledger, defining the financial relationship between physicians and hospitals. AB - OBJECTIVE: The purpose of the study was to determine the financial relationship between facility and professional revenue for care delivered by two orthopaedic surgeons in a Level I trauma center for patients with multiple orthopaedic injuries. DESIGN: Retrospective review of medical and financial records. SETTING: Level 1 trauma center. PATIENTS: Adult patients admitted with major multiple orthopaedic trauma were included if they had a significant pelvis/acetabulum injury associated with at least 2 extremity fractures, or if they had 3 or 4 extremities requiring surgical care. All patients had limited injury to other body systems, and all had completed their care related to the injury within the study period. We identified 68 patients who met the inclusion criteria. The mean Injury Severity Score was 22.6 (range, 9 to 57) and the mean New Injury Severity Score was 35.9 (range, 27 to 57). OUTCOME MEASURES: The trauma center has an integrated economy. The surgeons are employed by the hospital, and the information systems for facility and professional services are shared through a unified business structure. Inpatient hospital charges related to the initial trauma admission and subsequent hospital-based outpatient care were abstracted from the Trendstar billing system. All medical and financial records were reviewed to exclude inpatient hospital charges related to the delivery of care by nonorthopaedic services. Orthopaedic professional fees were abstracted from the Epic billing system used for physician services. RESULTS: The average facility charge was US$96,000, (range, $20,400 to $334,000, SD = $62,000) and the average orthopaedic professional charge was $24,900 (range, $5,200 to $60,300, SD = $14,000). The total facility charge for the study patients was $5,854,602 and the total orthopaedic professional charge was $1,516,568. The average orthopaedic trauma charge multiplier, the dollars of facility charge created by a single dollar of orthopaedic professional charge, was 3.86, (range, 1.35 to 15.2, SD = 3.0). When differences between collection rates were considered, the net revenue multiplier, the dollars collected by the hospital for facility services generated for each dollar collected by the orthopaedic surgeon, was 7.81. CONCLUSIONS: The majority of the charges and the net revenue related to the care of trauma patients fall on the hospital side of the ledger. The significant multiplier for orthopaedic care delivered in the inpatient setting increases the value of the orthopaedic traumatology service above and beyond the value of the labor component as reflected by the professional fee. Understanding the value of the multiplier in different clinical situations frames the interdependent relationship between physicians and hospitals. Changes in malpractice coverage, declining professional reimbursement and reported difficulties in securing on call coverage contribute to an emerging crisis in trauma care. The relationship between facility revenue and professional activity provides a firm basis to negotiate institutional support for orthopaedic traumatology. PMID- 18404030 TI - Economic value of orthopaedic trauma: the (second to) bottom line. AB - OBJECTIVES: We evaluated the economic aspects of an orthopaedic trauma section at a regional Level I, semi-academic community hospital. This study analyzes the economics of a dedicated hospital-based orthopaedic trauma program. METHODS: Institutional financial reports were analyzed for 2 time periods. In the pre program (PRE) period (2 years), we estimated the amount of forsaken revenue resulting from cases transferred to other institutions. In the post-program (POST) period (2 years), we analyzed financial reports to evaluate fiscal solvency. Health Care Cost and Utilization Project National Inpatient Sample (HCUP-NIS) data, International Classification of Diseases, 9th Revision (ICD-90 codes, and Eclipsys software were used. Standard accounting definitions for gross revenue, net revenue, direct costs, contribution margin, indirect costs, and net profit/loss were used. RESULTS: In the PRE-program period 88 patients were transferred; forsaken charges were about $1.25 million/year. Based on historic collection rates, there was about $450,000/year of actual lost revenue. In the POST-program period net revenue was about $7 million with a $1.5 million contribution margin, which increased 9%-11% in year 2. With inclusion of indirect costs, there was a net loss of nearly $5 million/year, but the financial software uses the direct cost expense as a major determinant of indirect costs. Based on the definition of indirect costs (overhead for lights, maintenance, etc) and with such expenses being used prior to the program, we felt that indirect cost was not an accurate variable and contribution margin is the better measure of economic value. CONCLUSION: We found that orthopaedic trauma is a financially viable program. Understanding the determination and interpretation of financial data is essential for any such analysis. PMID- 18404031 TI - The effect of an orthopedic trauma room on after-hours surgery at a level one trauma center. AB - PURPOSE: The purpose of this study is to examine the effect of establishing a dedicated operating room for unscheduled orthopedic cases and to evaluate a group of patients with isolated femur fractures. The frequency of after-hours surgery and the impact of patients who present with acute orthopedic injuries are reviewed. METHODS: A retrospective review of all orthopedic cases from the operating room scheduling system at a level-one trauma center was undertaken from October 2003 to September 2005. Before October 2004, unscheduled cases were placed on a shared add-on list, and no special priority was given to orthopedic cases. Additionally, a subset of adult patients with isolated femoral shaft fractures was identified to evaluate time from admission to surgery, operative time, frequency of transfer of care between surgeons, and total length of hospital stay. RESULTS: The number of orthopedic cases was 1799 in fiscal year 2004 (FY04) and 2046 in FY05, an increase of 14%. Overall, the hospital experienced an increase in level-one trauma activations from 1450 in FY04 to 1580 in FY05 (8.2%), and an increase in the number operative trauma cases from 447 to 494 (9.5%). Cases after 7:00 pm declined from 197 in FY04 to 165 in FY05, a decrease of 16%. Cases between midnight and 7:00 am declined from 63 in FY04 to 35 in FY05, a decrease of 44%. For the subset of femur fracture patients, transfer of care to another operating surgeon occurred 4.5 times more frequently. The median delay between admission and surgery increased from 5.7 hours to 10.9 hours. Median case duration increased from 106 to 127 minutes. CONCLUSIONS: It is possible to dramatically decrease the occurrence of after-hours orthopedic surgery in a level-one trauma center through the use of a dedicated room for unscheduled orthopedic trauma cases. Benefits include less frequent activation of after-hours operating room resources, fewer disruptions to the OR schedule and office hours, and more frequent fracture care by orthopedic traumatologists. The impact of a longer delay between admission and surgical treatment and more frequent transfer of care between surgeons deserves further evaluation. PMID- 18404032 TI - Orthopaedic faculty trauma call policies: a survey of accredited orthopaedic residency programs. AB - The purpose of the study was to determine the practices and policies for trauma call for orthopaedic faculty at residency training programs. A 2-page survey was mailed to the chairs of 141 nonmilitary, accredited residency programs. Responses were received from 106 (75% response rate). Of the responders, 97 (91.5%) of the programs were associated with a Level 1 trauma center. All faculty took trauma call in 44% of programs. The chair took trauma call in 60% of the programs. In 35% of programs, full-time faculty earned additional compensation for taking call. The source of this compensation for full-time faculty was the hospital alone in 72%. In 32 programs, a per-diem stipend (mean $696, range $100-1,500) was provided. In 59% of programs, there was a specific orthopaedic "trauma team" that took over patient care from other faculty members after call. Thirty-three percent of programs had a policy concerning age when a full-time faculty member went off trauma call. Of these, faculty came off call at age 50 years in 11% of programs; at age 55 years in 29%; at age 60 years in 40%; at age 65 years in 9%; and "other" in 11%. Twenty percent of chairs responded that trauma call adversely affected the chair's ability to recruit new faculty. There was a wide variety of policies concerning orthopaedic faculty trauma call. Additional studies on faculty trauma call are warranted. PMID- 18404033 TI - Biomechanical evaluation of clavicle fracture plating techniques: does a locking plate provide improved stability? AB - OBJECTIVES: To evaluate the biomechanical properties of both plate location (superior versus anterior-inferior) and plate type Small Fragment Contourable Dual Compression Plate (CDCP) versus 3.5 mm Universal Locking System Contourable Dual Compression Plate (Locked CDCP) in a synthetic midshaft transverse clavicle fracture model. METHODS: Twenty-four pre-osteomized synthetic clavicles were repaired with either CDCP or locked CDCP technology 3.5 mm plates in either the superior or anterior-inferior position to form 4 groups of 6 clavicles. These were subsequently tested to evaluate torsional and axial construct stiffness, as well as bending load to failure, bending failure stiffness, and method of failure. RESULTS: In axial compression, locked CDCP constructs were significantly more stiff than CDCP constructs (p < 0.001), but no statistically significant effect of plate location was observed. Torsional tests demonstrated a significant 2-way interaction favoring locked CDCP plates in the superior position and standard CDCP plates in the anterior-inferior position (p < 0.001). Bending failure testing revealed that the superior plate location had higher load to failure and bending failure stiffness than the anterior-inferior location (p < 0.0001). In addition, the superior locked CDCP plates demonstrated significantly greater bending failure stiffness than superior CDCP plates (p < 0.0001). CONCLUSIONS: Biomechanically, repairing a midshaft clavicle fracture with a superior plate was more favorable compared to anterior-inferior plating in terms of both load to failure and bending failure stiffness. Furthermore, superior locked CDCP plates show improved bending failure stiffness over superior CDCP plates. PMID- 18404034 TI - Comparison of three different treatment modalities in the management of humeral shaft nonunions (plates, unilateral, and circular external fixators). AB - OBJECTIVES: To compare 3 different fixation methods for the treatment of humeral shaft nonunions in terms of union time, functional outcome, and complications. DESIGN: Retrospective case series. SETTING: University hospital. PATIENTS: Between 1996 and 2004, 80 patients (mean age, 49; range, 15 to 86; 30 women and 50 men) with nonunions of the humeral shaft were treated surgically in our institution. Circular external fixators (CEF) were used in 35 patients, unilateral limb reconstruction system (LRS) fixators in 24 patients and fixation with plates in 21 patients. INTERVENTION: Surgical procedure included hardware removal in previously operated patients, autogenous grafting in all patients in the plate group and in those patients with atrophic nonunions in the external fixator groups, compression of the nonunion site in all patients. MAIN OUTCOME MEASUREMENTS: Radiological union time, complications, shortening, and disabilities of the arm, shoulder, and hand (DASH) score. RESULTS: Mean follow-up period was 48.1 months (range, 12 to 121). Mean radiological union time was 5.5 months (range, 1.5 to 12) in the CEF group, 5.2 months (range, 3 to 10) in the LRS group, and 5.7 months (range, 3 to 12) in the plate group. Mean DASH score was 23.7 in the CEF group, 18.6 in the LRS group, and 26 in the plate group. There were no statistical differences in terms of union time and the DASH score among the 3 groups. Successful union was achieved in 77 (96.3%) patients. CONCLUSION: Both external fixation and plate fixation produce excellent results in humeral shaft nonunions if applied properly. The procedure can be tailored to the surgeon's experience, keeping in mind that plate fixation demonstrates a longer healing time in those cases that had previous surgeries. PMID- 18404035 TI - Fit assessment of anatomic plates for the distal medial tibia. AB - OBJECTIVES: With the development and popularization of minimally invasive surgical methods and implants for fracture fixation, it is increasingly important that the available implants are precontoured to the specific anatomic location for which they are designed. The objective of this study was to develop a noninvasive method and criteria for quantifying the fit of a distal periarticular medial tibia plate and to test the method on a small set of tibia models. METHODS: The undersurface of the plate was extracted from a digital model of the plate. The surface of the plate was fitted to 21 computer tomography (CT)-based 3 dimensional (3-D) models of human tibiae. Four criteria were defined that constitute an anatomic plate fit and subsequently were applied for the quantitative fit assessment. The fitting of the plate undersurface to the bone was entirely conducted in a virtual environment. RESULTS: An anatomic fit of the plate was achieved for 4 of the models (19%). The individual categories generated fits of 62% (n = 13) for the proximal end; 43% (n = 9) for the proximal angle; 57% (n = 12) for the middle distance; and 57% (n = 12) for a distal fit. CONCLUSIONS: Although for the 4 individual criteria plate fits of 43%-62% were achieved, a global/anatomic fit only occurred for 19% of the bone models. This outcome is likely a result of bone morphology variations, which exist in a random population sample combined with the effects of a nonoptimized plate shape. Recommendations for optimizing the fit of the plate are discussed. PMID- 18404036 TI - Single- versus multiple-dose antibiotic prophylaxis in the surgical treatment of closed fractures: a meta-analysis. AB - OBJECTIVES: The use of prophylactic antibiotics in the surgical treatment of closed long bone fractures is well established. The duration and dosage of prophylaxis, however, vary significantly among surgeons. A systematic review and meta-analysis were performed to determine if multiple-dose perioperative antibiotic prophylaxis is more effective than a single preoperative dose in the prevention of surgical wound infections during the treatment of closed long bone fractures. DATA SOURCES: Articles were identified by searching the following medical databases: Medline, Medline In Process & Other Non-indexed Citations, Embase, CENTRAL, and the Cochrane Database of Systematic Reviews. Relevant conference proceedings and the reference section of selected manuscripts were also searched for additional studies. STUDY SELECTION: Studies were included if they were prospective randomized controlled trials of patients with closed fractures treated with surgical fixation or arthroplasty. The interventions must have directly compared a single preoperative prophylactic dose to a multiple-dose perioperative strategy. Studies were excluded if they involved open fractures. DATA EXTRACTION: The demographic information, prophylaxis strategy, wound infection rate, and risk ratio were extracted from each article. DATA SYNTHESIS: Seven trials and 3,808 patients were pooled using a random effects model. When compared to a regimen of multiple doses of prophylactic antibiotics, administration of a single preoperative dose has a risk ratio of 1.24 (95% CI 0.60-2.60). The pooled risk difference between the 2 strategies is 0.005 (95% CI 0.011-0.021). Neither result is significant. CONCLUSIONS: In the setting of closed long bone fractures, the pooled results failed to demonstrate superiority of multiple-dose prophylaxis over a single-dose strategy. The pooled estimates suggest that surgical wound infections are relatively rare events and that any potential difference in infection rates between prophylaxis strategies is likely quite small. However, because the confidence interval surrounding the pooled risk ratio spans 1.0 by such a large amount, we are unable to definitively recommend a preferred dosing regimen to prevent surgical wound infections. Although future research is required to ensure our prophylaxis decisions continue to be evidence based and cost-effective, it is unlikely that a single clinical trial will be able to provide the answer. The use of other quantitative methods, such as cost effectiveness analysis, may be helpful in modeling an optimal prophylaxis strategy. PMID- 18404037 TI - The subinguinal retroperitoneal approach for fractures of the acetabulum: a modified ilioinguinal approach. AB - The classic ilioinguinal approach is a standard procedure with reportedly high success rates in many displaced fractures of the acetabulum. Intraarticular visualization and exposure of the anterior wall and the quadrilateral plate are its main limitations. We propose a subinguinal approach based on the principle used for oncologic procedures that naturally require large exposures. The approach involves a retroperitoneal access below the inguinal ligament to preserve the integrity of the inguinal canal and allow ample exposure of anterior and medial wall fractures as well as the anterior hip capsule. Despite the apparent magnitude of the procedure, closure is fairly simple and anatomical because repair of the inguinal canal floor is not required. This modification may compensate for the limitations of the classic approach without additional risks or morbidities. PMID- 18404038 TI - Taylor Spatial Frame application with the aid of a fine wire half frame. AB - The Taylor Spatial Frame has become an important part of the trauma and reconstruction surgeon's armamentarium. We describe a technique to assist in the application of this device that does not hinder the use of the image intensifier or rely on an assistant to hold a constant position and aids provisional fracture reduction. PMID- 18404039 TI - One-stage shoulder and elbow arthroplasty after ipsilateral fractures of the proximal and distal humerus. AB - We report a case of 1-stage ipsilateral shoulder and elbow arthroplasty after comminuted fractures of both joints in a 77-year-old woman. The postoperative course was uneventful, and the patient showed prompt functional improvement and a good clinical result. Despite the higher risk of periprosthetic humeral fracture in cases of ipsilateral shoulder and elbow arthroplasty, the 1-stage procedure may be a promising and reliable solution for the treatment of complex upper limb fractures in elderly inactive patients. PMID- 18404040 TI - Brachial plexus palsy following proximal humerus fracture in patients who are skeletally immature. AB - From 1998 to 2006, 578 patients who were skeletally immature with proximal humerus fractures were treated at our institution. During that time period, 4 patients (0.7%) had associated brachial plexus and major peripheral nerve palsies. Average age at the time of injury was 12.3 years (range 10-14 years). Two fractures were physeal and 2 were metaphyseal. In all patients, the distal fracture fragment was displaced into the axilla, with resultant adduction and valgus malalignment at the time of injury. Two patients were treated with closed reduction and sling and swathe immobilization. One patient was treated with closed reduction and percutaneous pinning of the fracture. One patient was treated with sling immobilization without fracture reduction. All went on to bony healing with acceptable bony alignment. All patients were evaluated with serial physical examinations and radiographs to assess for neurologic recovery and bony healing. Average clinical and radiographic follow-up was 7.1 months (5-9 months). All patients demonstrated complete neurologic recovery by 5-9 months postinjury. All had neuropathic pain for at least 6 months after injury. No persistent neuropathic pain or functional limitations were seen at follow-up. Although rare, brachial plexus injury may accompany displaced proximal humeral fractures in patients who are skeletally immature. With careful attention to principles of fracture care, complete neurologic recovery may be expected within 9 months. PMID- 18404042 TI - Re: Traumatic segmental bone loss in a pediatric patient treated with bifocal bone transport. PMID- 18404043 TI - Re: Does medial tenderness predict deep deltoid ligament incompetence in supination-external rotation type ankle fractures? PMID- 18404044 TI - Ipsilateral femoral neck and shaft fracture in children. PMID- 18404045 TI - Modified abdominoplasty advancement flap for coverage of trauma-related hip disarticulations complicated by heterotopic ossification: a report of two cases and description of surgical technique. PMID- 18404046 TI - Recalcitrant distal humeral and proximal forearm nonunion: salvage using an extended pedicled radial forearm osseous flap. PMID- 18404047 TI - Posttraumatic cerebral infarction: incidence, outcome, and risk factors. AB - BACKGROUND: Outcome in patients with traumatic brain injury (TBI) is often affected by secondary insults including posttraumatic cerebral infarction (PTCI). The incidence of PTCI after TBI was previously reported to be 2% with no mortality impact. We suspected that recent advances in imaging modalities and treatment might affect incidence and outcome. We sought to define the incidence and mortality impact of PTCI. We also identified risk factors associated with PTCI. METHODS: We retrospectively reviewed all patients admitted between 2004 and 2006 with severe TBI (brain Abbreviated Injury Scale [AIS] score >2, Glasgow Coma Scale score [GCS] <9). Demographics, injury specifics, and clinical data were abstracted. All brain imaging studies were reviewed with an attending trauma radiologist. Statistical analysis of outcome data were performed using chi and Student's t test and multivariate analysis was performed using logistic regression to identify independent risk factors. RESULTS: Of the 384 patients identified with severe TBI; 93% sustained a blunt injury, 75% were men. Mortality was 21%, and 48% had a brain AIS score of 5. Mean age was 36 years (11-90 years), admission GCS score was 5 (3-8), and Injury Severity Score was 32 (9-75). Thirty one (8%) had a confirmed PTCI. The PTCI group had a significantly increased mortality (45% vs. 19%, p < 0.002), hospital length of stay (LOS) (25 days vs. 18 days, p < 0.02), and intensive care unit LOS (21 days vs. 15 days, p < 0.03). In multivariate analysis, sex, age, Injury Severity Score, Revised Trauma Score, admission GCS, and brain AIS were not associated with PTCI; whereas the presence of blunt cerebral vascular injury [odds ratio (OR) 4.0, 95% confidence interval (CI) 1.9-8.7], the need for craniotomy (OR 3.0, 95% CI 1.2-6.9), or treatment with recombinant factor VIIa (OR 3.1, 95% CI 1.1-8.0) were each independently associated with an increased risk of PTCI. CONCLUSIONS: The incidence of PTCI in patients with severe TBI is higher after severe brain injury than previously thought. PTCI has a significant impact on mortality and LOS. The presence of a blunt cerebral vascular injury, the need for craniotomy, or treatment with factor VIIa are risk factors for PTCI. Recognition of this secondary brain insult and the associated risk factors may help identify the group at risk and tailor management of patients with severe TBI. PMID- 18404048 TI - Epidemiology and 12-month outcomes from traumatic brain injury in australia and new zealand. AB - BACKGROUND: An epidemiologic profile of traumatic brain injury (TBI) in Australia and New Zealand was obtained following the publication of international evidence based guidelines. METHODS: Adult patients with TBI admitted to the intensive care units (ICU) of major trauma centers were studied in a 6-month prospective inception cohort study. Data including mechanisms of injury, prehospital interventions, secondary insults, operative and intensive care management, and outcome assessments 12-months postinjury were collected. RESULTS: There were 635 patients recruited from 16 centers. The mean (+/-SD) age was 41.6 years +/- 19.6 years; 74.2% were men; 61.4% were due to vehicular trauma, 24.9% were falls in elderly patients, and 57.2% had severe TBI (Glasgow Coma Scale score /=3 ventilator days), need for trachestomy, and mortality. Logistic regression was performed to identify factors that predicted pulmonary morbidity. RESULTS: Three hundred and eighty eight patients had >/=1 rib fracture. The mean (+/-standard deviation) age was 44 +/- 18. injury severity score was 21 +/- 11. Mortality was 6% (22 of 388). Sixty three (16%) patients developed respiratory failure. The mean number of rib fractures per patient was four (range, 1-23); 21% of patients had one rib fracture and 17% had six or more fractures. 208 (54%) of the initial CXRs were read as having no rib fractures. The mean number of rib fractures per patient in this group was 3.1 (CI95 2.9-3.2). In 43% (179 of 388) of patients, the CT radiology report incorrectly identified the number and location of the fractured ribs. Of these reports, 72% (129 of 179) differed from the prospective review by more than one fracture. The number of fractures was higher in patients who died (7 +/- 5 vs. 4 +/- 3; p = 0.02) and in those developing respiratory failure (6 +/ 4 vs. 3 +/- 3; p = 0.02). Any rib fracture or pulmonary contusion visible on the initial plain CXR significantly increased the incidence of pulmonary morbidity or mortality. CT determination of fracture location had no effect on respiratory failure, pneumonia, or mortality when fractures were confined to one anatomic location. The presence of rib fracture in more than anatomic region doubled the incidence of respiratory failure (24% vs. 12%; p = 0.002) but had no effect on mortality. Logistic regression identified only injury severity score and presence of a parenchymal injury on plain CXR as independent predictors of subsequent respiratory failure. CONCLUSIONS: Rib fracture mortality was lower than that in the previously published studies and is likely reflect the increased sensitivity of CT scan in diagnosing rib fractures. Screening CXRs miss rib fractures more than 50% of the time. Radiology reports are often not sufficiently descriptive or are incomplete with respect to the number and location fracture and reliance on these data will lead to erroneous conclusions. Using CT scanning, only the finding of rib fractures in multiple locations was associated with increased incidence of respiratory failure. In contrast, the presence of any parenchymal injury or visible rib fracture on the screening CXR significantly increases the risk for subsequent pulmonary morbidity (odds ratio, 3.8; CI95, 2.2-6.6). Although truncal CT scanning markedly improved the diagnosis and delineation of rib fractures, the screening CXR was a better predictor of subsequent pulmonary morbidity and mortality. PMID- 18404056 TI - Putting life on hold-for how long? Profound hypothermic cardiopulmonary bypass in a Swine model of complex vascular injuries. AB - BACKGROUND: Rapid induction of profound hypothermia for emergency preservation and resuscitation can improve survival from uncontrolled lethal hemorrhage in large animal models. We have previously demonstrated that profound hypothermia (10 degrees C) must be induced rapidly (2 degrees C/min) and reversed gradually (0.5 degrees C/min) for best results. However, the maximum duration of hypothermic arrest in a clinically relevant trauma model remains unknown. METHODS: Uncontrolled lethal hemorrhage was induced in 22 swine by creating an iliac artery and vein injury, followed 30 minutes later (simulating transport time) by laceration of the descending thoracic aorta. Through a thoracotomy approach, a catheter was placed in the aorta, and cold organ preservation solution was infused using a roller pump to rapidly induce profound hypothermia (10 degrees C) which was maintained with low-flow cardiopulmonary bypass. Vascular injuries were repaired during the asanguinous hypothermic low flow period. Profound hypothermia was maintained (n = 10-12 per group) for either 60 minutes or 120 minutes. After repair of injuries, animals were rewarmed (0.5 degrees C/min) and resuscitated on cardiopulmonary bypass, and whole blood was infused during this period. Animals were monitored for 4 weeks for neurologic deficits, organ dysfunction, and postoperative complications. RESULTS: The 4-week survival rates in 60- and 120-minute groups were 92% and 50%, respectively (p < 0.05). The surviving animals were neurologically intact and had no long-term organ dysfunction, except for one animal in the 120-minute group. The animals subjected to 120 minutes of hypothermia had significantly worse lactic acidosis, displayed markedly slower recovery, and had significantly higher rates of postoperative complications, including late deaths because of infections. CONCLUSION: In a model of lethal injuries, rapid induction of profound hypothermia can prevent death. Profound hypothermia decreases but does not abolish metabolism. With current methods, the upper limit of hypothermic arrest in the setting of uncontrolled hemorrhage is 60 minutes. PMID- 18404057 TI - A novel model of blunt thoracic aortic injury: a mechanism confirmed? AB - BACKGROUND: There is no consensus on the mechanism of traumatic injury to the thoracic aorta and no reproducible animal model. Advances in injury scene analysis suggest that lateral and oblique force vectors cause aortic injury. We hypothesized that the spectrum of aortic injury could be reproduced in an animal model by application of an obliquely directed load to the pressurized aorta. METHODS: Graded air impulses of 80, 100, 110, and 120 pounds per square inch (PSI) were delivered to the descending thoracic aorta of 19 swine with a novel pneumatic device. Aortic isthmus strain was recorded with microminiature probes. Gross and microscopic injury was recorded with digital photography. RESULTS: The spectrum of human aortic injury was reproduced in this model. Deep injuries to the aortic media were common. The majority of injuries occurred within the region of the isthmus. Impulse pressure of 120 PSI caused transections, whereas lower impulse pressure resulted in less severe injuries. Aortic isthmus strain was greater in the animals exposed to 120 PSI than those receiving lower PSI (19.6 +/ 4.9% vs. 8.7 +/- 2.5%, p = 0.067). CONCLUSIONS: Direct loading of the pressurized descending thoracic aorta causes isthmus injury secondary to aortic wall strain. Deep medial lesions are common and could propagate soon after injury to form pseudoaneurysms. A critical load is required to cause complete uncontained transection with exsanguination, which may have relevance to injury scene death. PMID- 18404058 TI - Surveillance bias and deep vein thrombosis in the national trauma data bank: the more we look, the more we find. AB - BACKGROUND: Deep vein thrombosis (DVT) has been identified as a marker of quality of care by various governmental and consumer groups. However, the lack of standardized DVT screening systems across trauma centers may introduce surveillance bias in the rates of DVT reported. We hypothesize that trauma centers with higher rates of duplex ultrasound detect more DVTs and subsequently report higher DVT rates to the National Trauma Data Bank. METHODS: We queried the National Trauma Data Bank version 6.1 and calculated ultrasound rates and DVT rates per trauma center. We excluded hospitals that did not report performing any ultrasounds or any complications. Simple and multiple linear regressions were used to describe the association between ultrasound and DVT rates among hospitals. RESULTS: One hundred forty-seven hospitals (16%) met the inclusion criteria, accounting for 578,252 patients (39% of the total patients in the dataset). When dividing hospitals into quartiles by duplex ultrasound rate, the DVT rate in the highest quartile was 7-fold higher than the average combined DVT rate in the first three quartiles (1.52% vs. 0.22%; p < 0.001). Multivariable analysis suggested that hospitals with an ultrasound rate /=10 units packed red blood cells in 24 hours of hospitalization). Multivariate logistic regression was used to develop prediction models. RESULTS: Seven US level I trauma centers (TC) enrolled 383 patients. 114 (30%) required MT. MT progressed rapidly (40% exceeded MT threshold 2 hours after TC arrival, 80% after 6 hours). One third of MT patients died. Two thirds of deaths were due to early exsanguination and two thirds of early exsanguination patients died within 6 hours. One third of the early MT survivors developed multiple organ dysfunction syndrome. MT could be predicted with standard, readily available clinical data within 30 minutes and 60 minutes of TC arrival (area under the receiver operating characteristic curve = 0.78 and 0.80). In patients who required MT, StO2 was the only consistent predictor of poor outcome (multiple organ dysfunction syndrome or death). CONCLUSION: MT progresses rapidly to significant morbidity and mortality despite level I TC care. Patients who require MT can be predicted early, and persistent low StO2 identifies those MT patients destined to have poor outcome. The ultimate goal is to identify these high risk patients as early as possible to test new strategies to improve outcome. Further validation studies are needed to analyze appropriate allocation and study appropriate use of damage control interventions. PMID- 18404071 TI - Prognostic factors in vitrectomy for posterior segment intraocular foreign bodies. AB - BACKGROUND: The aim of this study was to identify the prognostic factors influencing outcome after the removal of retained posterior segment intraocular foreign bodies (IOFBs) by pars plana vitrectomy. METHODS: We reviewed the records of 42 consecutive patients (39 men and 3 women with a mean age of 26.8 years) operated between January 2001 and January 2005 by the same surgeon for IOFB after penetrating ocular injury. All eyes underwent IOFB removal via pars plana vitrectomy. Postoperative retinal detachment was considered as the anatomic failure. Associations between anatomic outcome and various preoperative, operative, and postoperative variables were statistically analyzed. Chi-square test and Mann-Whitney U test were used to evaluate the association between two categorical variables. RESULTS: The mean time interval between the trauma and the IOFB removal was 5.3 days (range, 1 day to 30 days). The mean length of follow-up was 18.3 months (range, 6 months to 3 years). Preoperative retinal detachment was present in 19% of patients. After the IOFB removal, the retina was detached in 28.5% of patients (12 of 42 patients). After a second vitrectomy performed in 10 patients, final anatomic success rate was 89.8% (37 of 42 patients). Time between trauma and IOFB extraction, presence of intraocular hemorrhage, preoperative retinal detachment and primary surgical repair combined with the IOFB removal were significantly associated with the postoperative retinal detachment. Age, sex, entrance wound location, presence of endophthalmitis, location of IOFB, nature of IOFB, preoperative visual acuity, use of an encircling band, type of endotamponade, use of lensectomy were not significantly associated with the presence of postoperative retinal detachment. CONCLUSIONS: Delay in IOFB extraction, presence of intraocular hemorrhage, preoperative retinal detachment, primary surgical repair combined with IOFB removal are the predictive factors for anatomic failure. PMID- 18404072 TI - Evaluation and management of gunshot wounds of the penis: 20-year experience at an urban trauma center. AB - BACKGROUND: Although gunshot injuries to the penis occur relatively infrequently in patients with penetrating trauma, they often present dilemmas of subsequent evaluation and management. We review our extensive experience with gunshot wounds to the penis at a high volume urban trauma center. METHODS: The urologic trauma database was retrospectively reviewed to extract and compile information from the records of 63 patients treated for gunshot wounds to the penis. Data were accumulated for a 20-year period from 1985 to 2004 with regard to findings on physical examination, diagnostic evaluation, associated injuries, management, and outcome. We detail our technique of penile exploration and artificial erection in the management of these injuries. RESULTS: Penile gunshot wounds were associated with additional injuries in 53 of 63 (84%) patients. A total of 48 (76%) patients were taken to the operating room and 44 (70%) penile explorations were performed. Evaluation included retrograde urethrogram in 50 of 63 (79%) patients and was diagnostic for urethral injury in 11 of 12 (92%) cases. Primary urethral repair was performed in 8 of 12 (67%) patients with urethral injury versus 4 of 12 (33%) who underwent urinary diversion by means of suprapubic cystotomy. CONCLUSIONS: Evaluation and management of gunshot wounds to the penis may potentially be complex. Retrograde urethrogram should be performed in all cases except the most insignificant and superficial wounds. We describe our technique of penile exploration and artificial erection, noting excellent results in patients for whom follow-up is available. Additional studies are needed to prospectively evaluate techniques for management of gunshot urethral injuries. PMID- 18404073 TI - Management of colorectal injuries during operation iraqi freedom: patterns of stoma usage. AB - BACKGROUND: Management of penetrating colorectal injuries in the civilian trauma population has evolved away from diversionary stoma into primary repair or resection and primary anastomosis. With this in mind, we evaluated how injuries to the colon and rectum were managed in the ongoing war in Iraq. METHODS: The records of Operation Iraqi Freedom patients evacuated to National Naval Medical Center (NNMC) from March 2004 until November 2005 were retrospectively reviewed. Patients with colorectal injuries were identified and characterized by the following: (1) injury type; (2) mechanism; (3) associated injuries; (4) Injury Severity Score; (5) levels of medical care involved in patient treatment; (6) time interval(s) between levels of care; (7) management; and (8) outcomes. RESULTS: Twenty-three patients were identified as having either colon or rectal injury. The average ISS was 24.4 (range, 9-54; median 24). On average, patients were evaluated and treated at 2.5 levels of surgically capable medical care (range, 2-3; median 2) between time of injury and arrival at NNMC, with a median of 6 days from initial injury until presentation at NNMC (range, 3-11). Management of colorectal injuries included 7 primary repairs (30.4%), 3 resections with anastomoses (13.0%), and 13 colostomies (56.6%). There was one death (4.3%) and three anastomotic leaks (30%). Total complication rate was 48%. CONCLUSIONS: Based upon injury severity, the complex nature of triage and medical evacuation, and the multiple levels of care involved for injured military personnel, temporary stoma usage should play a greater role in military casualties than in the civilian environment for penetrating colorectal injuries. PMID- 18404075 TI - Cyclooxygenase-2 suppresses polymorphonuclear neutrophil apoptosis after acute lung injury. AB - BACKGROUND: Polymorphonuclear neutrophil (PMN) apoptosis is suppressed after acute lung injury (ALI), and strategies aimed at inducing PMN apoptosis are thought to be promising therapies for ALI. However, the mechanisms underlying PMN apoptotic suppression are unknown. Cyclo-oxygenase-2 (COX-2) has been shown to regulate tumor cell apoptosis and is up-regulated by inflammatory mediators in PMN. Therefore, we set out to determine whether up-regulation of COX-2 expression contributes to PMN apoptosis after ALI. METHODS: Experimental ALI was established in New Zealand rabbits by blunt chest trauma, and a correlation analysis of COX-2 immunohistochemical staining in lung tissue and PMN apoptosis in bronchoalveolar lavage fluid (BALF) was performed. Apoptosis was measured by flow cytometric analysis of annexin V and propidium iodide dual staining. As an in vitro correlate, normal PMNs were treated with BALF from injured lung (BALFALI) in the presence or absence of the COX-2 inhibitor, NS398. COX-2 mRNA levels and PMN apoptosis were then measured. RESULTS: PMN apoptosis was significantly decreased in BALF after injury. In contrast, COX-2 expression was significantly increased after injury. COX-2 protein expression and PMN apoptosis exhibited a strong inverse correlation (gamma = -0.75, p < 0.01). In vitro experiments revealed apoptosis of normal PMNs was significantly decreased by the addition of BALFALI. The addition of BALFALI was also associated with increased COX-2 mRNA levels. Treatment of cultures with NS398, 10 minutes before BALFALI addition, partially reversed all of these effects. CONCLUSIONS: Up-regulation of intrapulmonary COX-2 expression contributes to the suppression of PMN apoptosis after ALI. PMID- 18404074 TI - Effects of dimethyl sulfoxide, pyrrolidine dithiocarbamate, and methylprednisolone on nuclear factor-kappaB and heat shock protein 70 in a rat model of hemorrhagic shock. AB - BACKGROUND: Nuclear factor kappa B (NF-kappaB) is a transcription factor involved in the inflammatory response. Heat shock protein 70 (HSP70) is involved in the cell protection from various stresses. The aim of this study was to evaluate the effects of dimethyl sulfoxide (DMSO), pyrrolidine dithiocarbamate (PDTC), and methylprednisolone (MP) on liver, renal, and intestinal activation of NF-kappaB and HSP70 in a rat model of hemorrhagic shock (HS). METHODS: Sixty rats were randomized in 6 groups: sham-operated; only HS; HS and resuscitation with blood plus normal saline (NS); HS and resuscitation with blood/NS and 6 mg/kg DMSO; HS and resuscitation with blood/NS and 100 mg/kg PDTC; HS and resuscitation with blood/NS and 30 mg/kg MP. Rats were subjected to HS by blood removal to a mean arterial pressure of 35 to 40 mm Hg through the femoral artery. After 1-hour shock-period, the animals were resuscitated according to the experimental protocol. NF-kappaB and HSP70 expression in liver, kidney, and small intestine was analyzed 1 and 3 hours after resuscitation by immunohistochemistry. RESULTS: HS upregulated NF-kappaB activation and HSP70 expression (p < 0.05). Resuscitation was not associated with a further increase in NF-kappaB and HSP70 activation. DMSO, PDTC, and MP administration resulted in a decreased liver, renal, and intestinal activation of NF-kappaB associated with an increase of HSP70 expression (p < 0.05). CONCLUSIONS: Our results suggest that treatment with DMSO, PDTC, and MP can modulate the expression of NF-kappaB and HSP70 after HS in rats. This modulation may have potential effects in HS through inhibition of the NF-kappaB-dependent production of proinflammatory mediators. PMID- 18404076 TI - Could Beta blockade improve outcome after injury by modulating inflammatory profiles? AB - BACKGROUND: Cardioprotection with beta-receptor antagonists improves outcome in high risk patients undergoing elective surgery. Recent trials have demonstrated an association between beta blocker (BB) use and improved outcomes after injury. The mechanisms through which BB result in improved outcomes remain poorly elucidated. In vitro evidence supports that BB modulates the postinjury inflammatory response. The purpose of this study was to examine the effects of BB on inflammatory profiles in injured patients at increased risk for heart disease. METHODS: A pseudo-randomized, controlled trial of injured patients over 55 admitted to the intensive care unit was conducted. Patients were randomized to receive continuous BB or standard of care. Patients with a reported history of prehospital BB use were enrolled into an observational arm of the trial, continued on BB, and analyzed with the continuous BB group. Plasma interleukin (IL)-6 and IL-1beta levels were measured by enzyme-linked immunosorbent assay at baseline and day 1, 2, and 4 after BB initiation. Cytokine data were log transformed for normality assumptions. Repeated measures analysis of variance was used to test for within-group differences in cytokine levels over time. RESULTS: Forty-two patients were enrolled. Seventeen patients were randomized to the control group and 25 patients received continuous BB (10 randomized/15 observational). There was no difference in gender, age, prior history of heart disease, or admission heart rate, systolic blood pressure or initial base deficit between groups. Baseline levels of IL-6 and IL-1beta did not differ between groups. Levels of IL-6, but not IL-1beta, decreased over time in patients receiving BB (p = 0.04), whereas levels in controls remained unchanged (p = 0.27). There were no BB related complications. CONCLUSIONS: Use of BB decreases serum IL-6 levels over time in injured patients at risk for heart disease. This may contribute to improved outcomes noted in trauma patients receiving BB. Additionally, BB use in this population of patients is safe after endpoints of resuscitation have been met. PMID- 18404077 TI - Toll-like receptor 2 and 4 ligation results in complex altered cytokine profiles early and late after burn injury. AB - BACKGROUND: Toll-like receptors (TLR) 2 and TLR4 expressed on innate immune cells are important mediators of the immune response to pathogens. In this study, we hypothesized that burn injury results in altered cytokine secretion profiles after TLR2 or TLR4 ligation that is associated with altered TLR expression on innate immune cells. METHODS: Female C56BL/6 mice were subjected to 20% full thickness burn or sham injury. Three or 14 days after injury whole splenocytes or purified splenic macrophages were cultured with TLR2 ligand peptidoglycan or TLR4 ligand lipopolysaccharide. Supernatants were assayed for TNF-alpha, MCP-1, IL-6 and IL-10. Cell death was assessed using flow cytometry. Innate CD11b F4/80 macrophages were sorted 14 days after burn injury and TLR2 and 4 expression was determined by quantitative reverse-transcriptase polymerase chain reaction and flow cytometry. RESULTS: Burn injury results in a steady accumulation in the periphery of CD11bF4/80 macrophages. Macrophages purified early after burn injury upregulated TLR2 and 4, followed by a decrease of TLR2 and TLR4 expression late after burn injury. TLR2 and TLR4 ligation of an equivalent number of purified macrophages 3 days after burn injury revealed no significant differences in cytokine secretion compared with sham. Stimulation 14 days after burn injury revealed a significant reduction in tumor necrosis factor-alpha secretion by macrophages compared with sham mice. In contrast, interleukin-10 was significantly increased (mean, approximately 1.8-fold) late after burn injury after either TLR2 or TLR4 stimulation. Interleukin-6 and monocyte chemotactic protein-1 secretion was unchanged from sham levels. In contrast, whole splenocyte stimulation resulted in increased cytokine 3 days and 14 days after burn injury. This effect is likely caused by the accumulation of TLR macrophages, which are resistant to TLR-induced cell death. CONCLUSIONS: Cytokine secretion profiles after TLR2 and TLR4 ligation after burn injury are altered in a manner not clearly reflective of an anti-inflammatory or proinflammatory state and are associated with unique changes in the macrophage population. TLR2 and TLR4 ligation have complex and varied roles in mediating the immune response to burn injury. PMID- 18404079 TI - The evolution of blunt splenic injury: resolution and progression. AB - BACKGROUND: Nonoperative management of blunt splenic injury (BSI) has become the standard of care for hemodynamically stable patients. Successful nonoperative management raises two related questions: (1) what is the time course for splenic healing and (2) when may patients safely return to usual activities? There is little evidence to guide surgeon recommendations regarding return to full activities. Our hypothesis was that time to healing is related to severity of BSI. METHODS: The trauma registry at a level I trauma center was queried for patients diagnosed with a BSI managed nonoperatively between 2002 and 2007. Follow-up abdominal computed tomography scans were reviewed with attention to progression to healing of BSI. Kaplan-Meier curves were compared for mild (American Association for the Surgery of Trauma grades I-II) and severe (grades III-V) BSI. RESULTS: Six hundred thirty-seven patients (63.9% mild spleen injury and 36.1% severe injury) with a BSI were eligible for analysis. Fifty-one patients had documented healing as inpatients. Ninety-seven patients discharged with BSI had outpatient computed tomography scans. Nine had worsening of BSI as outpatients and two (1 mild and 1 severe) required intervention (2 splenectomies). Thirty-three outpatients were followed to complete healing. Mild injuries had faster mean time to healing compared with severe (12.5 vs. 37.2 days, p < 0.001). Most healing occurred within 2 months but approximately 20% of each group had not healed after 3 months. CONCLUSION: Although mild BSIs heal faster than severe BSIs, nearly 10% of all the BSIs followed as outpatients worsened. Close observation of patients with BSI should continue until healing can be confirmed. PMID- 18404078 TI - Contrast-enhanced ultrasound-guided microwave tissue coagulation therapy for hepatic trauma: an experimental study. AB - BACKGROUND: Nonoperative therapies for hepatic trauma have become more popular. Because there is a potential application of microwave tissue coagulation (MTC) in the minimally invasive management of hepatic trauma, this study aimed to evaluate the efficacy of contrast-enhanced ultrasound (CEUS)- guided MTC therapy for hepatic trauma. METHODS: Forty hepatic lacerations were created intraoperatively in 10 heparinized porcine livers. MTC therapy was performed under CEUS guidance. Two hours after MTC, CEUS and histopathologic examination were performed to assess the effect of MTC therapy. RESULTS: All hepatic lacerations with active bleeding were successfully controlled with MTC therapy. The bleeding times in grade II and III lacerations (192.0 +/- 108.0 seconds, 443.4 +/- 63.5 seconds, respectively) were longer than that in grade I (48.3 +/- 33.3 seconds; p < 0.01), and blood losses (24.8 +/- 6.5 g, 47.8 +/- 10.5 g) were also higher than in grade I (6.6 +/- 4.3 g; p < 0.01). All grade I lacerations (16 of 16, 100%), 7 grade II (7 of 16, 43.8%), and 3 grade III (3 of 8, 37.5%) were not enhanced on CEUS except for the electrode track appearing as a hyperechoic strip. In 9 grade II lacerations and 8 grade III lacerations, most of treated region was unenhanced apart from a few small regular vascular enhancements at the periphery of treated region. Histopathologic examination showed that there was no hematoma in any microwave-treated region and the hepatocytes in treated region were obviously necrotic. CONCLUSION: MTC therapy is a simple and effective method and may have great potential in minimally invasive treatment of hepatic trauma. PMID- 18404080 TI - QuikClot use in trauma for hemorrhage control: case series of 103 documented uses. AB - BACKGROUND: Local hemostatics have recently been introduced for field use to control external hemorrhage. The objective of this report is to describe the initial clinical experience with QuikClot, a zeolite that works by absorbing water and concentrating coagulation factors to stop bleeding in a series of patients. METHODS: Documented cases using a self-reporting survey sheet submitted by the users and first-hand detailed interviews with the users when possible. RESULTS: There were 103 documented cases of QuikClot use: 69 by the US military in Iraq, 20 by civilian trauma surgeons and 14 by civilian first responders. There were 83 cases involving application to external wounds and 20 cases of intracorporeal use by military and civilian surgeons. All field applications by first responders were successful in controlling hemorrhage. The overall efficacy rate was 92% with eight cases of ineffectiveness noted by physicians in morbid patients with massive injuries when the QuikClot was used as a last resort. These reported failures were thought to be a result of the coagulopathic state of the patient from massive resuscitation or the inability to get the product directly to the source of hemorrhage. When the QuikClot was applied on responsive patients, the heat generated by the exothermic reaction caused mild to severe pain and discomfort. There were three cases of burns caused by the heat generated by the QuikClot application with one case requiring skin grafting. There was one major complication from intracorporeal use caused by scar formation from a foreign body reaction. CONCLUSIONS: QuikClot has been effectively used by a wide range of providers in the field and hospital to control hemorrhage. PMID- 18404081 TI - Versatility of the free vastus lateralis muscle flap. AB - BACKGROUND: A flap that carries sufficient tissue with satisfactory pedicle length or diameter and with minor donor site morbidity is an ideal option for the reconstructive surgeon. Many flaps have been described with each one having specific advantages. The vastus lateralis (VL)-muscle flap is among them, but no special attention has been given to its valuable use. To evaluate and to propagate the versatility of the free VL-muscle flap, this flap was used in different pathology and in different anatomic areas. METHODS: Twenty-three patients with various defects after trauma and cancer therapy underwent reconstruction with 24 free VL-muscle flaps. RESULTS: The flap provided excellent reconstruction in all cases. All flaps were successful. Secondary operations included two debulking procedures, respectively, and one regrafting of parts of the flap because of partial skin graft loss. The donor site morbidity was minimal and no patient suffered from permanent partial limb dysfunction. CONCLUSIONS: The free VL-muscle flap is a versatile flap that offers specific advantages such as constant anatomy, ease of dissection in supine position, long pedicle length, large vessel diameter, good esthetic outcome, and minor donor site morbidity. This flap has proven to be most valuable also in difficult cases, including head and neck and lower limb reconstruction. PMID- 18404082 TI - Impediments to obtaining informed consent for clinical research in trauma patients. AB - BACKGROUND: Informed consent is required for, but a substantial barrier to, clinic research in trauma care. Exceptions have been established but remain controversial, and little objective data are available to illuminate this debate. METHODS: We prospectively assessed 2,011 consecutive patients admitted to the R Adams Cowley Shock Trauma Center of the University of Maryland Medical System for the potential to give consent for a hypothetical research trial within 1 hour of admission or whether consent could be obtained from a legally authorized representative (LAR) within 3 hours. These data were then linked with demographic, transport, physiologic, and injury data. RESULTS: Complete data were available for 1,734 patients: 982 (57%) appeared able to consent and 752 (43%) appeared unable to consent. Of the latter, LAR consent was potentially available for 404, leaving 348 (46%, 20% of all admissions) unconsentable. Those apparently able to consent were significantly less injured than those unable, but a third were subsequently found to have objective barriers to giving consent, and a further third had findings strongly suggestive of impairment. For those unable to consent, subsequent LAR consent was strongly associated with being a minor or being a woman. Lack of LAR consent was strongly associated with being the victim of intentional injury, with being an African-American male and-for European American men only-with being transported greater distances. CONCLUSIONS: Severely injured trauma patients can seldom provide consent for research studies, and LARs are often unavailable. Further efforts to develop workable mechanisms for exception from informed consent are justified. PMID- 18404083 TI - Research: questions and answers from academic trauma surgeons. AB - BACKGROUND: The impact of recent social and professional influences on trauma research is unclear. This study characterizes current research practices, opinions on research quality, and barriers to academic productivity, expressed by academic trauma surgeons. METHODS: A survey tool was administered electronically to members of the Eastern Association for the Surgery of Trauma. Questions on demographics, current and past research experience, perceptions of research quality trends, and barriers to academic success were included. RESULTS: Response rate was 40% (322 of 815). The mean age of respondents was 45 with 73% reporting completion of a critical care fellowship and 63% practicing in a university setting. The majority of respondents agreed or strongly agreed that both basic science (75%) and clinical (82%) research have become more difficult to perform. Greater difficulty in obtaining funding from their institutions was reported by 69% and by 61% for industry or private sources. Approximately 70% agreed that Institutional review board (IRB) regulations, confidentiality and consent requirements have impeded their research efforts whereas 86% agreed that increasing clinical requirements have inhibited their research efforts. Factors seen as impeding multi-institutional research, in order, were funding, IRB issues, poor coordination, commitment of investigators, and logistics. Perceived barriers to a successful research career were insufficient protected time (42%), funding (25%), personal motivation (11%), and IRB issues (11%). CONCLUSION: Research is viewed as being more difficult to conduct. The primary barriers to research productivity are perceived to be decreased protected time, decreased funding availability, and increased regulatory requirements. PMID- 18404084 TI - Diversion defended--military colon trauma. PMID- 18404085 TI - Tension band sutures using a washer for a proximal humerus fracture. AB - To prevent distraction and varus deformity between the humeral head and shaft, tension band sutures placed between the head of the interlocking screw and the rotator cuff, and we recommend using nonabsorbable sutures. We describe our simple procedure to overcome these difficulties in tension band suturing after reducing a proximal humerus fracture to maintain the reduction. PMID- 18404086 TI - One hundred consecutive splenectomies for trauma: is histologic evaluation really necessary? AB - Splenectomy remains the most commonly performed abdominal operation for trauma. Although the vast majority of these patients (pts) are young and healthy, histologic evaluation is still routinely performed. We propose that routine histologic sampling of an injured yet otherwise grossly normal spleen is unnecessary. A retrospective review of 100 consecutive pathologic specimens of pts undergoing splenectomy for trauma at a Level I trauma center was performed during a 25-month period. Data are reported as mean +/- SD. Average age was 34.4 +/- 15.1 years. There were 78 men and 22 women. Average injury severity score was 28.3 +/- 12.9. The most common mechanisms of injury were motor vehicle collisions (56%) and motorcycle collisions (15%). Average length of stay was 17.4 +/- 19 days. Microscopic pathologic findings returned as benign with no evidence for neoplasia in 99 of 100 specimens. Mean specimen weight was 184.6 +/- 188.7 g. Only one spleen, which appeared grossly abnormal and weighed 1,800 g, had abnormal histology demonstrating extramedullary hematopoiesis. This pt died before further workup. Our review suggests that the routine microscopic evaluation of spleens removed after traumatic injury, as is currently the standard of care at our institution, is unnecessary. Such examination should only be performed in cases of marked splenomegaly or in spleens that appear grossly abnormal. PMID- 18404087 TI - Damage control principles applied to penetrating neck and mandibular injury. PMID- 18404088 TI - Associated injury in pelvic fracture: colon herniation. PMID- 18404089 TI - Congenital anomalies presenting as acute orthopedic trauma. PMID- 18404090 TI - Procedural and clinical data plus electrocardiographic guidance greatly reduce the need for routine chest radiograph following central line placement. PMID- 18404092 TI - Biomechanics of volunteers subject to loading by a motorized shoulder belt tensioner. AB - STUDY DESIGN: A biomechanical study using human volunteers. OBJECTIVE: Motorized shoulder belt tensioning is a new seatbelt technology that is likely to be incorporated into future vehicles. The objective of this study was to characterize the upper torso biomechanics of 3 sizes of adult volunteers (5th percentile female, 50th percentile male, and 95th percentile male) subjected to motorized shoulder belt tensioning in the static environment. SUMMARY OF BACKGROUND DATA: There is a lack of volunteer data concerning the biomechanics of occupants subject to motorized precrash shoulder belt tensioning. Studies of torso repositioning by the air force for ejection seats are much too aggressive to be relevant to motorized systems. Low-level motorized shoulder belt tensioning is well tolerated by vehicle occupants but optimized performance by occupant size is unknown. METHODS: Nineteen male and 6 female subjects were instrumented in a fixture designed to support the occupant leaning forward and apply seatbelt tension. The subjects were 5th percentile females, 50th percentile males, and 95th percentile males. Reflective markers were placed on the subjects to monitor torso kinematics during tensioning. RESULTS: Changes in spinal curvature were small during shoulder belt tensioning and the angular motion of the torso originated within 4.2 cm of the pelvis-femur junction or H-point. Torso repositioning and retraction timing was found to be: 54.3 degrees in 0.78 seconds for the 5th percentile female, 57.6 degrees in 0.95 seconds for the 50th percentile male, and 42.2 degrees in 0.92 seconds for the 95th percentile male. CONCLUSION: Occupant size has a significant effect on retraction time to reposition the torso during shoulder belt tensioning. Larger vehicle occupants require more time because of a slower retraction velocity. The results are sufficiently simple that a lumped-mass model can predict tensioning kinetics. PMID- 18404093 TI - How does a three-dimensional continuum muscle model affect the kinematics and muscle strains of a finite element neck model compared to a discrete muscle model in rear-end, frontal, and lateral impacts. AB - STUDY DESIGN: A finite element (FE) model of the human neck with incorporated continuum or discrete muscles was used to simulate experimental impacts in rear, frontal, and lateral directions. OBJECTIVE: The aim of this study was to determine how a continuum muscle model influences the impact behavior of a FE human neck model compared with a discrete muscle model. SUMMARY OF BACKGROUND DATA: Most FE neck models used for impact analysis today include a spring element musculature and are limited to discrete geometries and nodal output results. A solid-element muscle model was thought to improve the behavior of the model by adding properties such as tissue inertia and compressive stiffness and by improving the geometry. It would also predict the strain distribution within the continuum elements. METHODS: A passive continuum muscle model with nonlinear viscoelastic materials was incorporated into the KTH neck model together with active spring muscles and used in impact simulations. The resulting head and vertebral kinematics was compared with the results from a discrete muscle model as well as volunteer corridors. The muscle strain prediction was compared between the 2 muscle models. RESULTS: The head and vertebral kinematics were within the volunteer corridors for both models when activated. The continuum model behaved more stiffly than the discrete model and needed less active force to fit the experimental results. The largest difference was seen in the rear impact. The strain predicted by the continuum model was lower than for the discrete model. CONCLUSION: The continuum muscle model stiffened the response of the KTH neck model compared with a discrete model, and the strain prediction in the muscles was improved. PMID- 18404094 TI - Long-term functional outcome in pyogenic spinal infection. AB - STUDY DESIGN: Retrospective. OBJECTIVE: To assess long-term adverse functional outcome following pyogenic spinal infection using standardized outcome measures, Oswestry disability index (ODI), and medical outcomes study short form-36 (SF 36). SUMMARY OF BACKGROUND DATA: There is minimal published data regarding the long-term functional outcome in pyogenic spinal infection. Previous studies have used heterogeneous, unreliable, and nonvalidated measure instruments yielding data that is difficult to interpret. METHODS: All cases of pyogenic spinal infection presenting to a single institution managed operatively and nonoperatively from 1994 to 2004 were retrospectively identified. Follow-up was by clinical review and standardized questionnaires. Inclusion in each case was on the basis of consistent clinical, imaging, and microbiology criteria. RESULTS: Twenty-nine cases of pyogenic spinal infection were identified. Twenty-eight percent were managed operatively and 72% with antibiotic therapy alone. Nineteen patients (66%) had an adverse outcome at a median follow-up of 61 months, despite only 5 patients (17%) having persistent neurologic deficit. A significant difference in SF-36 physical function scores was observed between patients with adverse outcome and patients who recovered (P = 0.003). SF-36 scores of all patients, regardless of management or outcome, failed to reach those of a normative population. A strong correlation was observed between ODI and SF-36 physical function scores (rho = 0.61, P < 0.05). Seventeen percent (n = 5) of admissions resulted in acute sepsis-related death. Delay in diagnosis of spinal infection (P = 0.025) and neurologic impairment at diagnosis (P < 0.001) were significant predictors of neurologic deficit at follow-up. Previous spinal surgery was associated with adverse outcome in patients requiring readmission within 1 year of hospital discharge following first spinal infection (P = 0.018). No independent predictors of adverse outcome, persistent neurologic impairment, readmission within 1 year, or acute death were identified by logistical regression analysis. CONCLUSION: High rates of adverse outcome detected using SF 36 and ODI suggest under-reporting of poor outcome when American Spinal Injury Association score alone is used to qualify outcome. PMID- 18404095 TI - Diabetes and perioperative outcomes following cervical fusion in patients with myelopathy. AB - STUDY DESIGN: Database study using the Nationwide Inpatient Sample administrative data from 1988 through 2004. OBJECTIVE: To examine perioperative morbidity and mortality for patients diagnosed with myelopathy, with and without diabetes mellitus (DM) (and subclassifications) following cervical spinal fusion. SUMMARY OF BACKGROUND DATA: DM has been associated with worse outcomes in a variety of orthopedic procedures including spinal surgery. Evidence that patients with DM have more complications following cervical fusion, specifically those treated for myelopathy, has been suggested within the literature but has been poorly explored. METHODS: Data from 37,732 patients within Nationwide Inpatient Sample database (1988-2004) with diagnostic codes specifying the presence of myelopathy and who underwent cervical fusion were included in the analysis. Patients were compared on the basis of the presence of DM, type of DM, and whether DM was controlled or uncontrolled. Bivariate statistical analyses compared postoperative complication rates while multivariate statistics were used to determine likelihood of complications with DM. RESULTS: Multivariate regression modeling outlined higher likelihoods of complications and hospital discharge variables with DM, particularly if it was diagnosed as uncontrolled disease. Fewer significant discrepancies in complications were noted in comparison of Type I versus Type II DM. CONCLUSION: This nationally representative study of inpatients in the United States provides evidence that patients with DM who received cervical fusion secondary to myelopathy are associated with greater perioperative complications, nonroutine discharge, and increased total charges. Subanalyses suggest that uncontrolled DM is a significant associative factor in outcome. PMID- 18404096 TI - Threshold cervical range-of-motion necessary to detect abnormal intervertebral motion in cervical spine radiographs. AB - STUDY DESIGN: Whole human cadaver model to assess a common diagnostic test for cervical spine stability. OBJECTIVE: Determine criteria that can be used to determine if sagittal plane angular motion of the head/neck during a cervical spine flexion/extension study is adequate to reliably assess intervertebral motion (IVM). SUMMARY OF BACKGROUND DATA: Flexion/Extension radiographs of the cervical spine are commonly used to help identify specific abnormalities in IVM. Several authors have recognized that inadequate patient effort can make flexion/extension studies unreliable, but validated guidelines for assessing the adequacy of these studies are not available. METHODS: Increasingly severe anterior-to-posterior (N = 6), and posterior-to-anterior (N = 6) soft tissue injuries were simulated in the cervical spines of 12 human cadavers. Sagittal plane radiographic images were taken with 4 gradually increasing amounts of overall flexion and extension motion of the head. IVM was measured for each level of sagittal plane rotation of the head/neck using previously validated computer assisted methods. RESULTS: With less than 60 degrees of sagittal plane rotation of the head/neck, intervertebral rotation or displacement was almost never greater than the 95% confidence interval previously established for asymptomatic people. Even with 60 degrees or more motion, intervertebral rotation and displacement were within normal limits after extensive damage to the soft tissues. The center-of-rotation was the most sensitive measure for detecting soft tissue damage. CONCLUSION: The results of this study suggest that clinicians should make sure patients can flex and extend their head/neck to a minimum range of 60 degrees before evaluating them for a dynamic motion study to assess cervical spine stability. Even with adequate motion, interverterbral rotation and translation can remain within normal limits in the presence of extensive soft tissue damage. The most sensitive measure for detecting soft tissue damage was center-of-rotation although it lacks specificity, particularly in the presence of underlying degenerative changes, and is not readily assessed in most clinical situations. PMID- 18404097 TI - Delayed migration of a screw into the gastrointestinal tract after anterior cervical spine plating. AB - STUDY DESIGN: Case report and clinical discussion. OBJECTIVE: To describe a rare case of spinal cervical screw migration into the gastrointestinal tract 11 years after surgery. SUMMARY OF BACKGROUND DATA: Anterior cervical spine fusion and stabilization with plating is a well-established procedure for cervical myelopathy, cervical spinal trauma, and spinal infectious disease. Esophageal injury has been related to screw or plate extrusion. METHODS: We present a 45 year-old white man suffering from severe quadriparesis and neck pain. Cervical spine magnetic resonance images showed spondylodiscitis and spinal fracture of C4 and C5 vertebral bodies. He underwent anterior surgical decompression consisting of C4 and C5 corpectomies and fusion with fibular allograft fixated with a cervical plate. The patient's neurologic examination gradually improved during his follow-up. RESULTS: The patient returned 11 years after cervical spine graft and plating, complaining of severe dysphagia and high fever. Radiographs of the cervical spine showed a screw back-out. Three days later, a new radiograph of the cervical spine revealed a progression of the screw's extrusion. In a new cervical spine radiograph, obtained immediately before surgical intervention, the displaced screw was no more visible. Abdominal radiograph showed the missed screw in the right lower abdominal quadrant. Barium meal swallowing test showed no leakage on the cervical area. At 6 months follow-up, the patient is in good condition and symptom free. CONCLUSION: In this case, the delayed esophageal perforation occurred 11 years after initial surgery: the rapid progression of the screw extrusion after initial pull-out (6 days) was documented by several radiographs and spontaneous closure of the fistula was achieved. PMID- 18404098 TI - Compartment syndrome with acute renal failure: a rare complication of spinal surgery in knee-chest position. AB - STUDY DESIGN: Case report of a patient who presented with compartment syndrome with acute renal failure after surgery for lumbar disc prolapse in knee-chest position. OBJECTIVE: To describe an unusual complication of compartment syndrome in lower limb with acute renal failure following discectomy in knee-chest position. SUMMARY OF BACKGROUND DATA: Compartment syndrome and acute renal failure following lumbar discectomy in knee-chest position has been described in only 2 cases previously. METHODS: Case report and review of literature. RESULT: Subsequent to prolonged surgery in knee-chest position this patient developed compartment syndrome with acute renal failure. Immediate fasciotomy was done, resulting in good recovery. CONCLUSION: Spine surgeon performing discectomy in knee-chest position should be aware of such an unusual complication in the postoperative period, especially if the surgery gets prolonged. Time and position should be monitored carefully to avoid such a serious complication. Early diagnosis and urgent intervention in the form of fasciotomy is required for good results. PMID- 18404100 TI - Cell therapy using articular chondrocytes overexpressing BMP-7 or BMP-10 in a rabbit disc organ culture model. AB - STUDY DESIGN: Rabbit knee articular chondrocytes overexpressing human growth factors were injected into cultured intervertebral disc explants. Survival of the injected cells and accumulation of extracellular matrix were assessed. OBJECTIVE: To define the utility of cell-based gene delivery approach for repair of the intervertebral disc. SUMMARY OF BACKGROUND DATA: Back pain associated with symptomatic disc degeneration is a common clinical condition. Growth factors stimulate disc cell metabolism, but the ideal method for in vivo delivery has not been established. Cells as a vehicle for delivering growth factors to the disc offer potential advantages, including prolonged production of the growth factor within the disc and vital cells to participate in the repair process. METHODS: New Zealand white rabbit articular chondrocytes transduced with adenovirus expressing human bone morphogenetic protein-7 and green fluorescence protein (GFP) (AdhBMP-7), human bone morphogenetic protein-10 and GFP (AdBMP-10), or GFP alone (AdGFP, as a control) were injected into whole disc explants. Discs were maintained in culture for 1 to 2 months. At the conclusion of the culture periods, cell survival was assessed by fluorescence microscopy and extracellular matrix accumulation was assessed with biochemical methods. RESULTS: Chondrocytes achieved long-term survival in the cultured disc explants. The discs treated with chondrocytes/BMP-7 demonstrated a 50% increase in proteoglycan content within the nucleus pulposus compared to control (chondrocytes/GFP), while discs injected with chondrocytes/BMP-10 failed to show a significant increase in proteoglycan accumulation. CONCLUSION: Our study demonstrates the ability of transduced articular chondrocytes to survive and promote proteoglycan accumulation when transplanted into the intervertebral disc. These data support the potential of a cell-based gene therapy approach for disc repair. Further studies using this approach in animal models are indicated as a step towards achieving disc repair in humans. PMID- 18404101 TI - Plasmin-matrix metalloproteinase cascades in spinal response to an experimental disc lesion in pig. AB - STUDY DESIGN: Proteinases were immunohistochemically stained to analyze degenerated discs and paradiscal tissues in comparison to contiguous control tissues in an experimental porcine model of intervertebral disc degeneration. OBJECTIVE: The aim was to analyze plasmin and metalloproteinases known to participate in mutual activation cascades. SUMMARY OF BACKGROUND DATA: Comparison of the degenerated discs and paradiscal structures with control tissues disclosed accumulation of plasmin and induction of matrix metalloproteinases (MMP), MMP-1 and MMP-2 in the discs, but some other MMPs in reactive and remodeling tissues. MATERIAL AND METHODS: In 6 domestic pigs, the cranial L4 endplate was perforated to penetrate the nucleus pulposus. Three months later, the animals were killed and the experimental and the contiguous control vertebrae, complete with their intervertebral discs, were excised and subjected to histologic and immunohistochemical examinations. RESULTS: Immunohistochemical analysis disclosed increased expression of MMP-1 and MMP-2 in the traumatized and degenerated intervertebral discs. Some MMPs were also induced in all paradiscal structures (bone marrow, vertebral bone, and spinal ligaments), or decreased in already scarred areas. The common denominator for all the anatomic sites studied was accumulation of plasmin. CONCLUSION: Fibroblast collagenase (MMP-1) and gelatinase A (MMP-2), capable of degrading native and denatured collagen, were induced in degenerating intervertebral discs. Use of an experimental model enabled demonstration that biomechanical destabilization and degeneration of the disc also affects all other paradiscal structures, which are subjected to proteolysis and/or reparative fibrosis apparently representing remodeling of the spine subjected to pathologic stress. Profiling of various MMPs and plasmin, known to participate in mutual activation cascades, suggests that plasmin could activate pro-MMP-1, pro-MMP-2, pro-MMP-3, pro-MMP-7, pro-MMP-9, and pro-MMP-13 and alone or/and in cooperation with MMP-3 initiate at least 2 mutual MMPs activation cascades driven by activated MMP-3 and MMP-7. PMID- 18404102 TI - Age-related effects of cocultured rat nucleus pulposus cells and macrophages on nitric oxide production and cytokine imbalance. AB - STUDY DESIGN: A study of age-related effects on nitric oxide (NO) and cytokine production in cocultured rat nucleus pulposus (NP) cells and macrophages. OBJECTIVE: To evaluate the effects of age on NO and cytokine production in an in vitro model of cocultured NP cells and macrophages. SUMMARY OF BACKGROUND DATA: It is well known that the clinical characteristics of lumbar disc herniation differ with age. The relationship between age-related differences in clinical features and immuno-chemical factors, such as NO and inflammatory cytokines, has not been established. METHODS: Male Sprague Dawley rats (n = 45), including 15 animals from 3 groups (3-, 12-, and 32-weeks old), were used. NP cells and exudated peritoneal macrophages were cocultured in serum-free media. NO levels were measured at 2-, 24-, 48-, and 72 hours using the Griess method. After 7 days of culture, the production of cytokines, including tissue inhibitor metalloproteinase-1, interferon-gamma (IFN-gamma), and interleukin-10 (IL-10) were evaluated. RESULTS: NO levels of coculture increased with age. In the coculture groups, tissue inhibitor metalloproteinase-1 and IFN-gamma level of 3 weeks old were statistically higher than 12 and 32 weeks old. IL-10 level of 3 weeks old was statistically lower than 12 and 32 weeks old. CONCLUSION: NO levels of cocultures increased with age that suggests inflammatory reactions increase with age. This study showed an age-related cytokine imbalance, as represented by levels of IFN-gamma and IL-10. Stress and aging are thought to affect the extracellular matrix and change the immunologic response. Younger rat NP cells had higher cell-mediated immunity activity, while the older rat had higher humoral immunity activity. These results demonstrate that age affects the immunologic response attributable to NP cells. Further studies are needed to elucidate the mechanism of this newly observed occurrence and to apply these findings clinically. PMID- 18404103 TI - Puncture of a lumbar intervertebral disc induces changes in spontaneous pain behavior: an experimental study in rats. AB - STUDY DESIGN: Changes in spontaneous behavior was studied in rats after a controlled puncture of a lumbar intervertebral disc. OBJECTIVE: To study if puncture of a lumbar disc would induce recordable changes in spontaneous pain behavior. SUMMARY OF BACKGROUND DATA: Anular tears are common and may be found both in patients with low back pain and in asymptomatic patients. It has been suggested that anular injury may relate to low back pain either by stimulation of local sensory receptors in the posterior part of the anulus fibrosus or by ingrowth of newly formed nerve fibers into the deeper parts of the disc. The objective of the study was to analyze if a controlled puncture of a lumbar intervertebral disc might induce recordable changes in spontaneous behavior of rats. METHODS: After anesthesia, the L4-L5 disc was punctured in 10 rats. Ten other rats received sham surgery. Spontaneous behavior was assessed at days 1, 3, 7, 14, and 21 after surgery. RESULTS: Statistically significant differences in behavior were seen at all days analyzed. Most consistent were increases in "grooming" and in "wet-dog shakes." CONCLUSION: Puncture of a lumbar intervertebral disc in the rat produces changes in spontaneous behavior mainly seen as increased "grooming" and "wet-dog shakes," 2 behaviors that have been suggested to indicate stress and pain. PMID- 18404104 TI - An intact facet capsular ligament modulates behavioral sensitivity and spinal glial activation produced by cervical facet joint tension. AB - STUDY DESIGN: In vivo experiments using a rat model of painful facet joint distraction. OBJECTIVE: To determine whether tension of the facet capsular ligament is requisite for producing pain for joint loading and to define effects on spinal glial activation. SUMMARY OF BACKGROUND DATA: Cervical facet joint loading may initiate pain for certain conditions. While facet capsule tension has been proposed as requisite for pain, this hypothesis has not been tested. METHODS: Using an established rat model of painful C6-C7 distraction, tension was applied after transection of the left facet capsule; the right capsule remained intact. Each rat (n = 8) received the same distraction simultaneously applied across both the intact and cut capsules. Sham procedures were performed on separate rats (n = 4) with no joint distraction. Bilateral forepaw mechanical allodynia was measured as a pain outcome. Cervical spinal cord tissue (C7) was harvested on day 7 to detect glial reactivity using immunohistochemistry. RESULTS: Distraction mechanics were consistent with conditions eliciting persistent behavioral hypersensitivity. Allodynia was produced for an intact capsule and was significantly elevated over both the cut capsule (P < 0.004) and sham (P < 0.002). Transecting the capsule before distraction did not produce elevated allodynia, except on day 7. Spinal astrocytic reactivity paralleled allodynia; glial fibrillary acidic protein expression for an intact capsule was significantly greater than the cut and sham responses (P < 0.04), with no difference observed between the cut and sham spinal astrocytic reactivity. Spinal microglial activation did not differ between groups. CONCLUSION: Results suggest ligament tension may be required to produce pain from facet joint loading. Further studies of other cellular responses are needed to define the mechanisms of painful facet joint injury. PMID- 18404105 TI - Comparison of human mesenchymal stem cells derived from adipose tissue and bone marrow for ex vivo gene therapy in rat spinal fusion model. AB - STUDY DESIGN: Rat spinal fusion model. OBJECTIVE: To compare the efficacy of human adipose tissue-derived mesenchymal stem cells (HATDMSCs) and human bone marrow-derived mesenchymal stem cells (HBMDMSCs) transduced with an adenovirus containing the cDNA for bone morphogenetic proteins (BMP)-2 for inducing spinal fusion in an athymic rat model. SUMMARY OF BACKGROUND DATA: Recombinant BMPs have successfully induced spinal fusion in clinical trials. However, large doses are required for adequate bone repair. Regional gene therapy may deliver proteins to specific anatomic sites more efficiently. Gene transfer techniques using HATDMSCs have recently been tested. METHODS: Spinal fusion was performed in rats with different treatments: Group I (n = 10) collagen sponge containing HATDMSCs transfected with adeno-BMP-2, Group II (n = 10) collagen sponge containing HBMDMSCs transfected with adeno-BMP-2, Group III (n = 10) collagen sponge containing recombinant BMP-2 (10 mug), Group IV (n = 6) collagen sponge containing HATDMSCs transfected with adeno-LacZ, Group V (n = 6) collagen sponge containing HBMDMSCs transfected with adeno-LacZ, and Group VI (n = 6) collagen sponge alone. Radiographs were obtained at 4, 6, and 8 weeks. After sacrifice, the rat spines were assessed by manual palpation, microcomputed tomography, and histologic analysis. RESULTS: At 8 weeks, spinal fusion was observed in all Groups I, II, and III rats. 75% (15 of 20) of the gene therapy treatment animals (Groups I and II rats) had spontaneous extension of the fusion to a second level. No Groups IV, V, and VI rats developed fusion. New bone volume was significantly greater in Groups I and II than in Group VI. CONCLUSION: HATDMSCs transfected with adeno-BMP-2 induce abundant bone formation and have a similar posterolateral spinal fusion in rats as similarly genetically modified HBMDMSCs. Both are potential strategies for spinal fusion and may be a more efficient method of obtaining spinal fusion over currently used grafting substances. PMID- 18404106 TI - Coculture of bone marrow mesenchymal stem cells and nucleus pulposus cells modulate gene expression profile without cell fusion. AB - STUDY DESIGN: Changes in gene expression profile and cell fusion of mesenchymal stem cells (MSC) and nucleus pulposus cells (NPC) after coculture were analyzed. OBJECTIVE: To investigate the mechanisms of the interaction between NPC and MSC such us differentiation, stimulatory effect, and cell fusion. SUMMARY OF BACKGROUND DATA: Introduction of exogenous cells to supplement and replenish intervertebral disc cell population offers a potential approach to treat intervertebral disc degeneration (IDD). Recent evidences showed that intradiscal injection of MSC effectively alter the course of IDD in vivo, and the regenerative potential may result from up-regulated extracellular matrix protein synthesis mediated by MSC and NPC interaction. METHODS: Using a double labeling cell system and flow activated cell sorting, we quantitatively analyzed changes in the gene expression profile of human male MSC and female NPC after coculture in a 3-dimensional system that allows short distance paracrine interactions typical of the nucleus pulposus. Furthermore, we analyzed for cell fusion in the cell interaction by fluorescence in situ hybridization (FISH) for X and Y chromosomes, using a 3-dimensional culture system to allow cell-to-cell interactions conducive to cell fusion. RESULTS: Two weeks of coculture cell interaction in a 3-dimensional environment induces a change in MSCs towards a more chondrogenic gene expression profile indicating MSC differentiation, and NPC gene expression changes in matrix and chondrogenic genes demonstrating only a modest trophic effect of MSC on NPC. Moreover, FISH analysis demonstrated that cell fusion is not responsible for MSC plasticity in the interaction with NPCs. CONCLUSION: This study clarifies the mechanism of MSCs and NPCs interaction in a 3-dimensional environment, excluding cell fusion. These data support the use of undifferentiated MSC for stem cell therapy for IDD treatment. PMID- 18404107 TI - Early histologic changes following polymethylmethacrylate injection (vertebroplasty) in rabbit lumbar vertebrae. AB - STUDY DESIGN: An ex vivo histologic study in rabbits. OBJECTIVE: To evaluate the early histologic effects of polymethylmethacrylate (PMMA) injection on bone and intraosseous neural tissue following vertebroplasty in rabbit lumbar vertebrae. SUMMARY OF BACKGROUND DATA: Vertebroplasty with PMMA is performed to treat painful osteoporotic vertebral fractures. Early pain relief has been consistently documented, but its mechanism has not been elucidated. Among the mechanisms of pain relief may be the immediate stabilizing effects of the cement, and the exothermic reaction during curing, which may lead to intraosseous neural ablation. It has been well established that PMMA can induce thermal osteonecrosis after arthroplasty, but the potential for osteonecrosis after vertebroplasty has not been established. Previous studies have suggested that temperature elevations during cement curing may induce thermal bone necrosis. However, this cause-and effect relationship has not yet been histologically studied in an animal model. METHODS: Vertebroplasty with PMMA was performed at 2 levels in 12 New Zealand rabbits (24 levels); trochar insertion without PMMA injection was performed at 3 levels each of 2 control animals (6 levels). Sacrifice was performed 24 hours after the procedure. Histologic examination was performed to evaluate the presence of bone or intraosseous neural tissue necrosis. RESULTS: Half of the levels with PMMA showed evidence of necrosis at the bone-cement interface. Almost all (11 of 12) showed only focal necrosis, with only 1 specimen showing necrosis along the entire periphery of the PMMA. The other 12 specimens and all control levels displayed no bone necrosis. There was no evidence of intraosseous neural tissue necrosis in control or PMMA-injected specimens. CONCLUSION: Injection of PMMA in rabbit lumbar vertebral bodies produces early, focal bone necrosis in only half of cases, suggesting that competency of the cement-bone interface is reasonable in most cases. No evidence of intraosseous neural tissue damage was found. PMID- 18404108 TI - A biomechanical study of artificial cervical discs using computer simulation. AB - STUDY DESIGN: A virtual simulation model of the subaxial cervical spine was used to study the biomechanical effects of various disc prosthesis designs. OBJECTIVE: To study the biomechanics of different design features of cervical disc arthroplasty devices. SUMMARY OF BACKGROUND DATA: Disc arthroplasty is an alternative approach to cervical fusion surgery for restoring and maintaining motion at a diseased spinal segment. Different types of cervical disc arthroplasty devices exist and vary based on their placement and degrees of motion offered. METHODS: A virtual dynamic model of the subaxial cervical spine was used to study 3 different prosthetic disc designs (PDD): (1) PDD-I: The center of rotation of a spherical joint located at the mid C5-C6 disc, (2) PDD II: The center of rotation of a spherical joint located 6.5 mm below the mid C5 C6 disc, and (3) PDD-III: The center of rotation of a spherical joint in a plane located at the C5-C6 disc level. RESULTS: A constrained spherical joint placed at the disc level (PDD-I) significantly increased facet loads during extension. Lowering the rotational axis of the spherical joint towards the subjacent body (PDD-II) caused a marginal increase in facet loading during flexion, extension, and lateral bending. Lastly, unconstraining the spherical joint to move freely in a plane (PDD-III) minimized facet load build up during all loading modes. CONCLUSION: The simulation model showed the impact simple design changes may have on cervical disc dynamics. The predicted facet loads calculated from computer model have to be validated in the experimental study. PMID- 18404109 TI - Quantitative anatomy of subaxial cervical lateral mass: an analysis of safe screw lengths for Roy-Camille and magerl techniques. AB - STUDY DESIGN: Determination of lateral mass screw lengths with Roy-Camille and Magerl techniques of screw insertion using computerized tomography in 98 young, asymptomatic North American volunteers. OBJECTIVE: To provide reliable and normative data on safe screw lengths using the Roy-Camille and Magerl techniques of lateral mass fixation in the subaxial cervical spine. SUMMARY OF BACKGROUND DATA: Lateral mass screw lengths have been studied in the past using differing subject and measurement characteristics and small sample sizes. Results demonstrated considerable variation in screw length and influencing factors. Inappropriate screw lengths can result in neurovascular injury during screw insertion, facet joint damage, or inadequate fixation. METHODS: Bicortical screw lengths were bilaterally measured at each spinal level from C3-C7 in 98 young volunteers using computed tomography reconstructions through the lateral masses obtained in the plane of the screw in Roy-Camille and Magerl techniques. RESULTS: With both techniques, trajectories were longest at C4-C6, shorter at C3, and shortest at C7. Screw lengths were greater in males when compared with females at all levels. Average Magerl screw lengths were approximately 2.6 mm longer at C3 C6 levels, and approximately 1.3 mm longer at the C7 level when compared with Roy Camille technique. There was minimal correlation between screw lengths and anthropometric measurements including stature, body weight, and neck length. CONCLUSION: Significant variations exist at each subaxial level with either technique. We recommend the surgeon determine screw lengths for fixation at each level using preoperative sagittal oblique computed tomography scans, which provide the most accurate technique of preoperative templating for screw length. PMID- 18404110 TI - Facet asymmetry in normal vertebral growth: characterization and etiologic theory of scoliosis. AB - STUDY DESIGN: The shape and orientation of the thoracic and lumbar zygapophyseal facets at the T1-L5 level in children were measured and analyzed. OBJECTIVE: To detect the pattern of zygapophyseal facet asymmetry in the thoracic and lumbar spines in children. SUMMARY OF BACKGROUND DATA: Whereas many studies have defined the pattern of zygapophyseal facet asymmetry in adults, there is insufficient data in children. METHODS: A 3-dimensional digitizer was used to measure zygapophyseal facet size, topography (length, width, concavity, convexity, and lateral interfacet height), and orientation (transverse and sagittal facet angles) at the T1-L5 level. Thirty-two complete, nonpathologic skeletons of children (age range from 4 to 17 years), housed at the Hamman-Todd Human Osteological Collection (Cleveland Museum of Natural History, Cleveland, OH) were assessed. Statistical analysis included paired t tests and analysis of variance. RESULTS: In general, zygapophyseal facet asymmetry in children exists only in the superior facets of the thoracic spine and is independent of age: The right superior facet is significantly shorter than the left in all thoracic vertebrae T1-T12 (up to -2.91 mm at T1), and significantly wider than the left in thoracic vertebrae T1-T9 (T8 excluded) (P < 0.003). The right superior transverse and sagittal facet angles are significantly greater than the left in thoracic vertebrae T1-T11, indicating a lesser inclination (in the sagittal plane) and more frontally positioned facet (in the transverse plane) (P < 0.003). Facet asymmetry was not evident in the superior or inferior facets of the lumbar vertebrae. CONCLUSION: Facet asymmetry in thoracic vertebrae appears in early childhood. The pattern of this asymmetry differs from that reported for adults and may be considered as a possible contributing etiological factor in the development of different types of idiopathic scoliosis. PMID- 18404111 TI - Clinical course and significance of the clear zone around the pedicle screws in the lumbar degenerative disease. AB - STUDY DESIGN: A retrospective study. OBJECTIVE: To report the clinical course and significance of postoperative radiolucent zones around pedicle screws. SUMMARY OF BACKGROUND DATA: Radiolucent zones around pedicle screws have been considered to indicate loss of fixation, delayed union, or pseudarthrosis. Few reports have described the course and significance of them. METHODS: Plain radiographs were evaluated from 190 patients with pedicle screw fixation. When 1 mm or greater circumferential lucency was confirmed around a screw on plain radiographs from 2 or more directions, the patient was judged as clear-zone positive. The course of clear-zone positivity was investigated and the relationships between clear-zone positivity and the following items were investigated: (1) number of levels fused, (2) bone union, (3) posterolateral lumbar fusion versus posterior lumbar interbody fusion, (4) clinical results, (5) bone mineral density, and (6) screw types. RESULTS: Although clear zones were observed in 78 patients 6 months after surgery, the clear zone-positive rate decreased over time and the progression of bone union and the number of patients decreased to 28 at the final follow-up 3 years or more after surgery. Interobserver and intraobserver coefficient of variability in clear-zone judgment was over 0.86, and agreement was over 96%. At final follow-up, pseudarthrosis was found in 15 patients. Although the predictive value of the positive test for the presence of clear zones for pseudarthrosis at final follow-up was 0.50, sensitivity was 0.93 and specificity was 0.92. There were significant differences in the clear zone-positive rate in the number of levels fused and bone mineral density among the investigated items. CONCLUSION: As approximately two thirds of the clear zones disappeared over time, the presence of clear zones did not necessarily mean pseudarthrosis. However, clear zones persisting for 2 years or longer after surgery are a great risk of pseudarthrosis. Therefore, careful observation of clear zones around pedicle screws is of great significance as an evaluation of bone union. PMID- 18404112 TI - Pain in the back and neck are with us until the end: a nationwide interview-based survey of Danish 100-year-olds. AB - STUDY DESIGN: Nationwide interview survey. OBJECTIVES: To determine the 1-month prevalence and impact of back and neck pain among centenarians and to investigate associations between back and neck pain and other health measures such as physical function, depression, comorbidity, and self-rated health. SUMMARY OF BACKGROUND DATA: To our knowledge prevalence and impact of back and neck pain has never been studied among the oldest old. METHODS: Information on the 1-month prevalence of back and neck pain and bothersomeness of back and neck pain was collected using face-to-face interviews in a nationwide survey of Danish centenarians. In addition, information on physical and mental functional abilities, self-rated health, and comorbidities were collected. RESULTS: Two hundred fifty-six persons completed the interview (response rate, 56%). Twenty nine percent of women and 17% of men had experienced back pain, and 23% of women and 19% of men had experienced neck pain during the past month. This was comparable to another nationwide Danish survey of younger seniors. Twenty-one percent had been bothered by back pain either when moving, resting, or sleeping. Poor overall physical function, bad self-rated health, and higher depression score were associated with higher prevalence of back and neck pain. CONCLUSION: Back pain and neck pain continue to be common and bothersome complaints even into extreme old age. Thus, back pain and neck pain are not limited to working populations. PMID- 18404113 TI - The use of expertise-based randomized controlled trials to assess spinal manipulation and acupuncture for low back pain: a systematic review. AB - STUDY DESIGN: Systematic review. OBJECTIVE: To assess current use of expertise based randomization in trials of acupuncture or spinal manipulation for low back pain. SUMMARY OF BACKGROUND DATA: The randomized clinical trial is often referred to as the gold standard for providing evidence to guide therapeutic decisions. Random allocation of participants to intervention and control groups theoretically should balance these groups for both known and unknown prognostic factors; however, when randomizing patients to competing interventions in which the clinician's skill is a central aspect of the intervention, (e.g., surgery, chiropractic, rehabilitation) a differential expertise bias may exist if a majority of clinicians participating have greater expertise in 1 of the 2 interventions under evaluation. Randomizing patients to therapists experienced in the interventions under investigation can overcome this bias. METHODS: We systematically identified relevant randomized controlled trials published up to December 2005. Two independent reviewers extracted data in duplicate using a standardized form. RESULTS: Of 12 eligible trials, none made use of an expertise based randomized trial design. CONCLUSION: Investigators designing acupuncture or spinal manipulation trials in which 2 or more active therapies are compared should consider expertise-based randomization to increase the validity and feasibility of their efforts. PMID- 18404114 TI - Vertebroplasty using real-time, fluoroscopy-controlled, catheter-assisted, low viscosity cement injection. AB - STUDY DESIGN: Case series. OBJECTIVE: Evaluation of new vertebroplasty technique. SUMMARY OF BACKGROUND DATA: Viscous polymethylmethacrylate cement is frequently used to avoid leakage in vertebroplasty. However, the large force required to inject the cement can require the use of special equipment or the direct injection of the cement without a cannula. In addition, injection under fluoroscopic guidance with a short or absent cannula is associated with a higher intraoperative radiation dose. A method using an angiographic catheter for thin cement injection is presented. METHODS: Real-time, fluoroscopy-controlled, catheter-assisted, thin cement injection was used for 102 vertebroplasty sessions (84 patients, 148 vertebrae). Leakage was monitored in 85 procedures via computed tomography. RESULTS: All vertebral bodies were successfully and satisfactorily filled with bone cement. Cement leakage was evident in 50% of the 85 asymptomatic cases (25 paraspinal, 29 epidural, and 30 intradisc space). On follow-up, new compression fractures were noted in the same (n = 5) or adjacent (n = 16) vertebral body, and elsewhere (n = 10). Vertebroplasty provided pain relief in >90% of cases. Recurrent compression fracture occurred in 5 vertebrae of 4 patients after vertebroplasty. New compression fractures occurred in 16 adjacent vertebral bodies of 11 patients and in 10 vertebral bodies remote from the site. One patient who developed osteomyelitis was successfully treated with antibiotics alone. CONCLUSION: Real-time, fluoroscopy-controlled, catheter-assisted, low viscosity cement injection is a safe and feasible method for vertebroplasty, and produces a comparable outcome as high-viscosity cements. PMID- 18404115 TI - Historical perspective: the development and use of spinal disease categories. AB - This historical perspective traces the history of the development of pathoanatomically based spinal disease diagnosis and the subsequent use of these diagnoses for scientific and social purposes. Spinal diagnostic categories have been used positively to guide both clinical (including their use as the primary units of investigation in many epidemiological and evidence-based projects) and basic science (studying the pathoanatomic disease outside of the patient) research programs--the aim being the improvement of patient care and outcomes. They are also used socially to confer/reject acceptability to patient behavior; to justify health policy decisions; to structure medical relationships; to shape medical/institutional infrastructure; to direct patient care via guideline establishment; and to manage health care. The positive and negative implications of the use of spinal disease categories are discussed. PMID- 18404116 TI - [Bilateral Duane syndrome associated with crocodile tears, and congenital megacolon: a case study]. AB - Duane's syndrome is a congenital form of strabismus characterized by horizontal eye movement limitation and specifically by globe retraction with palpebral fissure narrowing in attempted adduction. It is usually associated with ocular and systemic pathology. We report a case of 2-year-old girl presenting Duane syndrome associated with crocodile tear syndrome, preauricular tags, and Hirschsprung disease. Through this rare case, we discuss different pathogenic hypotheses of this syndrome and associated abnormalities. PMID- 18404117 TI - [Setting the preferential retinal locus. Part 1. Analysis of the rehabilitation results as a function of positioning]. AB - INTRODUCTION: The aim of this study was to compare the various positions of preferred retinal locus (PRL) with the results of low vision rehabilitation to reading in view to elucidate the mechanisms involved. MATERIAL AND METHODS: We studied 100 visually impaired subjects who came to the center for rehabilitation of low vision. The technique is based on a scanning laser ophthalmoscope recording, compared with angiofluorographic pictures to obtain the PRL position with respect to the fovea. RESULTS: It is shown that unsuccessful rehabilitations are not related to the eccentricity or to the angular value of the PRL but are only correlated to its absence. The difficulties that arise from the multiplicity of the PRL or from the confinement of the fovea by an annular scotoma can be overcome with the help of rehabilitation. CONCLUSION: The upper and right positions are manifestly dominant, but they do not favor reading or writing. We believe that their purpose is to favor the integration of the new visual message of visually impaired people to facilitate their daily life activities. Therefore, using one or another of these peexisting PRL during eccentric viewing training should be preferred. PMID- 18404118 TI - [Aberrometry and contrast sensitivity after cataract surgery: aspherical IOL evaluation]. AB - PURPOSE: We report a prospective clinical study investigating efficacy in terms of aberrometry and contrast sensitivity of the aspherical IOL (SW60AT) versus the standard IOL Alcon (SN60AT). PATIENTS AND METHODS: Twenty patients in each group underwent phacoemulsification of both eyes. Both eyes of each patient received an aspherical IOL (SW60AT) or a similar standard IOL (SN60AT). The integrity of ocular function was assessed with clinical examination before surgery. Postoperative examination (1 month after the second surgery) included best visual acuity, an aberrometry with a 5-mm pupil (Bausch and Lomb), and a contrast vision (Zalonghi scale) in scotopic place examination. RESULTS: We noted no statistically significant difference between the two groups before surgery. Best spectacle-corrected visual acuity improved in both groups with no difference (8.15/10+/-1.39/10 and 7.95/10+/-1.54/10 for the aspheric group eyes and 7.85/10+/-2.13/10 and 7.60/10+/-2.14/10 for the standard group eyes). Otherwise, a statistically significant difference was found for the Z40 aberration with RMS=0.0095 with SW60AT versus RMS=0.07625 with SN60AT. The pupils had a similar size in both groups. There were no statistically significant differences for contrast sensitivity between two IOLs; nevertheless, contrast vision seemed to be better with the aspherical lens (33.15+/-6.85 points and 32.15+/-6.95 points for the standard group eyes and 36.05+/-3.63 points and 35.70+/-3.93/10 points for the aspheric eyes). CONCLUSION: The quality of vision can be objectively increased with the aspherical IOL because of less spherical aberration. PMID- 18404119 TI - [Multiple evanescent white dot syndrome and acute idiopathic blind spot enlargement: angiographic and electrophysiologic findings]. AB - Multiple white evanescent white dot syndrome (MEWDS) and acute idiopathic blind spot syndrome enlargement (AIBSE) are two acute retinopathies occurring in young patients: improvement is the rule. The present review describes two cases: one case of MEWDS and one of AIBSE. We observed these patients from the acute phase until absolute recovery. Comparing these two reports, we noted differences in ophthalmoscopic, fluorescein angiographics, and multifocal electroretinographic (mERG) analysis, but common findings were observed in clinical presentation and indocyanine green imagery. The first and recent description of these two disorders dates from less than 25 years ago. Although the pathophysiology is still unknown, some authors suggest that these two syndromes could be two different expressions of the same disease, depending on the patient's sensitivity. In our two case reports, multifocal ERG anomalies were directly related to visual impairment and visual field alterations. MEWDS and AIBSE are two acute retinopathies that must be recognized, because of the spontaneous visual recovery that the patient needs to understand for a good follow-up. PMID- 18404120 TI - [Central corneal thickness in a pediatric population using a noncontact specular microscope: a study of 405 cases]. AB - AIM: Central corneal thickness has become very important in the interpretation of intraocular pressure. The aim of this study was to correlate pachymetry values in a pediatric population with age, sex, and refraction. To our knowledge, the scientific literature has not reported such information. PATIENTS AND METHOD: This prospective study was conducted from May 2005 to November 2006 and included 405 eyes (186 eyes in boys and 219 eyes in girls) in children undergoing a general pediatric ophthalmologic visit. Patient age ranged from 3 to 16 years; three age groups were formed: 3-4 years, 5-9 years, 10-16 years. Objective refraction with cycloplegic was measured using a Nidek ARK 700 A autorefractometer and three refraction groups were formed. Pachymetry measures were taken using the Topcon SP 2000P specular microscopy. Exclusion criteria were a history of ocular surgery, lens wearers, and any disease that could alter the cornea. RESULTS: The mean age was 8.21 +/- 3.55 years, the mean refraction was +1.24 +/- 4.02 D. The mean central pachymetry was 529 +/- 32 microm: 534 +/- 31 microm in the male group and 524+/-31 microm in the female group, establishing a difference in pachymetry with no differences in refraction or age between the two groups. In the refraction groups, we found a significant difference in pachymetry between emmetropic (523+/-33 microm) and hyperopic (533 +/- 31 microm) groups and between hyperopic and myopic groups (520 +/- 32 microm) (p=0.005). There was no difference between emmetropic and myopic groups (p=0.902), nor in pachymetry between age groups (p=0.086). DISCUSSION: Except for differences found in refraction groups, the results reported herein were similar to those found with the ultrasonic method (the reference method), but they had been obtained under general anesthesia in most studies. CONCLUSION: Even though this is not the reference method of pachymetry measures, it seems more appropriate for children, because it is reliable, the measurements are taken quickly, they are noninvasive, and they do not cause the child undue stress (noncontact method). PMID- 18404121 TI - [Transpupillary thermotherapy of choroidal melanomas]. AB - PURPOSE: To evaluate the effectiveness of transpupillary thermotherapy (TTT) as a single treatment of choroidal melanomas. METHODS: We studied a series of 50 cases of choroidal melanoma treated with TTT and evaluated them with standardized A scan and B-scan echography before and after treatment (1 week, 1, 3, and 6 months after treatment, then every 6 months). RESULTS: There were 22 males and 28 females with a mean age of 57 years (range, 22-78). The mean thickness of the lesion was 2.7 +/- 0.6 mm (range, 1.63-3.72). The mean follow-up was 38 months (range, 21-41). In almost all the eyes treated with TTT, substantial regression of tumoral microcirculation was observed after 1 week associated with a 70%-80% reduction in tumor thickness after 6 months (stabilized during follow-up). In one case of juxtapapillary melanoma with a thickness of 3.72 mm, another TTT application was necessary for local relapse. Visual acuity (VA) decreased to 20/30 in two cases (4%) and to 20/200 in four cases (8%) after the development of a cystoid macular edema. These latter patients were treated with two intravitreal injections (range, 1-3) of triamcinolone acetonide, and after a follow-up of 25 months (range, 21-29) VA improved to 20/20 in the first two cases, while two of the four other cases improved to 20/40 and two to 20/30. CONCLUSION: Transpupillary thermotherapy has visual results similar to those obtained with irradiation for the treatment of choroidal melanomas. Nevertheless, considering the high rate of recurrence at 5 and 10 years after transpupillary thermotherapy alone, most authors think that transpupillary thermotherapy is useful when associated irradiation for the treatment of choroidal melanomas, but its precise indications need to be defined. PMID- 18404122 TI - [Efficacy of omega-6 essential fatty acid treatment before and after photorefractive keratectomy]. AB - PURPOSE: Following PRK, a regression or the appearance of haze is often observed. This type of problem may be caused by an anomalous response by the stroma, probably mediated by keratocytes. Controlling keratocyte apoptosis therefore affects the outcome and stability of PRK. Inhibiting the transmission of the apoptosis signal from the damaged corneal epithelium to the keratocytes attenuates cell activation. This can be achieved by using a systemic product that will control the release of the mediators of the inflammation and stimulate tear production. The aim of this prospective, randomized study was to test the effects of a supplement containing omega-6 fatty acids (linoleic and (-linolenic acid), ( carotene, group B vitamins and trace elements (copper, zinc), when administered both before and after PRK. METHODS: Eighty subjects undergoing PRK were randomly divided into two groups. The group treated with omega-6 fatty acids included 18 females and 22 males, with a mean age of 32 years; the control group comprised 20 females and 20 males (mean age, 30 years). Statistical evaluation of the results was carried out on Schirmer test and fluorescein staining values and the state of the epithelium. RESULTS: In the Schirmer tests, a comparison between the two groups of patients who underwent surgery (groups 1 and 2) showed a statistically significant difference after 7, 15, and 30 days' treatment in favor of the group treated with omega-6 fatty acids. The values relating to the fluorescein staining test and the state of the epithelium showed a statistically significant difference in favor of the group treated with omega-6 fatty acids. CONCLUSION: These results confirm the real advantages of using omega-6 essential fatty acids to optimize and stabilize the outcome of PRK. PMID- 18404123 TI - [Primary exotropia: importance of cerebral MRI]. AB - INTRODUCTION: Primary exotropia is a divergent strabismus that appears from the first day of life to the second year. It can be isolated or associated with a pathological context requiring cerebral imaging to determine diagnosis and prognosis. The objective of this study was to report the sensorimotor state and the result of MRI in infantile exotropia. PATIENTS AND METHOD: Forty-seven children with primary exotropia had a complete ophthalmologic assessment (visual acuity, binocular vision, refraction with cycloplegia, eye fundus) and cerebral MRI (average age, 16 months). Deviation was classified into four categories<4 degrees, > or =4<10 degrees, > or =10<20 degrees, > or =20 degrees. MRI results were classified into seven categories: white matter abnormalities (gliosis, delay of maturation, periventricular leukomalacia, aspecific hyperintense signal, and necroses); Virchow-Robin enlargement space and enlarged subarachnoid space; gray matter abnormalities (necroses, cerebral atrophy, occipital cortex, basal ganglia); ventriculomegaly; thin corpus callosum; cerebellar injury; and tumor. RESULTS: There was a statistically significant increase in the rate of pathological MRI as the angle deviation increased: 76.6% of patients had a pathological ophthalmologic exam (amblyopia, ptosis, head posture, Duane's syndrome, cataract, albinism, or pigmentary retinopathy). We found 38% nystagmus and 38% optic nerve hypoplasia. There was a real pathological context in 61.7% of the exotropia cases: 27.7% prematurity, 31.9% fetal distress, 21.3% facial dysmorphy (plagiocephaly), 12.8% psychomotor delay, and 14.9% epilepsy. Only three children had isolated exotropia. Thirty-four cerebral MRI (72.3%) were not normal. In pathological MRI, there was 61.8% white matter injury, 41.2% gray matter injury, and 17.6% thin corpus callosum. CONCLUSION: This study demonstrates the fundamental contribution of cerebral MRI in infantile exotropia. The greater the deviation, the more abnormal the MRI results are. The pathological context and ophthalmological abnormalities are important in infantile exotropia. PMID- 18404124 TI - Retinal myelinated nerve fibers associated with macular pseudohole. AB - We report the case of a 24-year-old woman who presented unilateral extensive peripapillary myelinated nerve fibers associated with macular pseudohole and bilateral low myopia. Visual acuity, slit lamp biomicroscopy, automated perimetry, fundus color photography, and optical coherence tomography (Stratus OCT) were performed. We discuss the clinical findings and diagnostic details of unilateral extensive peripapillary myelinated nerve fibers associated with macular pseudohole. PMID- 18404125 TI - [Ophthalmomyiasis interna: two case studies]. AB - Myiasis is a pathology caused by the presence of fly larvae in the human organism. Ophthalmomyiasis interna is characterized by the presence of the parasite within the eye but also in the eyelids and the orbit. Herein, the authors present the clinical cases of two patients presenting ophthalmomyiasis interna revealed by retinal detachment for the first patient and by panuveitis for the second patient. These two cases should make ophthalmologists aware of this rare disease that can effect patients living in direct contact with ovine races. PMID- 18404126 TI - [Hypophysial macroadenoma revealed by clinical symptoms simulating chronic glaucoma]. AB - INTRODUCTION: A large optic cup is a sign of glaucoma, although it has been reported in other optic neuropathies, particularly in compressive neuropathies. CASE REPORT: A 37-year-old man presented with a decrease in visual acuity for 2 months. Fundus biomicroscopic examination revealed a glaucoma-like bilateral and asymmetric large optic cup. Neuroradiological examination revealed an expansive mass in the sella turcica that was compressing the optic tract. CONCLUSIONS: Optic nerve compression can lead to perimetric alterations and optic disc anomalies such as those seen in normal-tension glaucoma. Age under 50 years, visual loss, and neuroretinal rim pallor may lead to a suspicion of compressive neuropathy. PMID- 18404127 TI - [Imaging the ocular surface with confocal microscopy: histology without samples]. AB - Developed in the 1990s, confocal microscopy contributed a new way of viewing corneal structures at levels close to those of histology. However, these systems were expensive, difficult to handle and use, had poor resolution, and were not usable for more superficial or peripheral structures. These devices were therefore restricted to research and were not used routinely in clinical practice. New-generation devices such as the HRT Rostock Cornea Module recently provided a new approach with high-resolution, fully digital images of the entire ocular surface, including limbus and conjunctiva. These images are close to those obtained ex vivo using histological procedures. The technique is painless and minimally invasive. Confocal microscopy should therefore become a very helpful means to explore ocular surface diseases in the near future. PMID- 18404128 TI - Treatment of conjunctival squamous neoplasias with interferon alpha 2ab. AB - The classical approach to the treatment of squamous neoplasias of the ocular surface is based on surgical resection and cryotherapy. High rates of recurrence have been demonstrated if the margins are not free after the resection. Primary or adjuvant chemotherapy with mitomycin C (MMC) or 5-fluorouracil has been employed to treat these neoplasias, but severe side effects on the ocular surface have been described. Interferon (INF) alpha 2b is less toxic to the ocular surface. The experience in the treatment of conjunctiva-cornea intraepithelial neoplasia (CIN) with topical or intralesional INF alpha 2b is based on isolated cases or very short series. In this report, we review the published studies and include our personal experience. The safety and effectiveness of INF alpha 2b in the treatment of primary and recurrent cases of CIN are described. The absence of serious side effects after topical administration of INF alpha 2b leads to the recommendation of this modality of therapy for primary and recurrent cases of CIN. PMID- 18404129 TI - [Intraocular lens power calculation after keratorefractive surgery]. AB - The number of keratorefractive procedures designed to correct refractive errors has dramatically increased over the last few years. The techniques for cataract extraction and intraocular lens implantation have evolved into a refractive surgical procedure as well as an operation to improve best corrected visual acuity and/or spectacle independence. The calculation of intraocular lens power for a desired refractive target can be challenging in post-refractive surgically treated eyes, given the frequent case reports of "refractive surprises" after cataract surgery. After corneal refractive surgery, the direct use of the measured topographic or keratometric values, with no correction, results in less accurate calculation of intraocular lens (IOL) power required for cataract surgery than calculation in virgin eyes. After laser refractive surgery for myopia, this could result in an overestimation of the corneal power and subsequent underestimation of the IOL power, therefore leading to a hyperopic outcome after phacoemulsification. Conversely, after laser refractive surgery for hyperopia, inaccuracy in the keratometric power estimation could result in a myopic outcome after phacoemulsification. Despite current progress in this subject, awareness of the shortcomings of classical methods and suggested strategies to improve accuracy can be valuable to clinicians. This article provides an overview of the possible sources of error in intraocular lens power calculation in post-keratorefractive patients, and reviews the methods to minimize intraocular lens power errors. PMID- 18404130 TI - [Bilateral elastosis of the limbus]. AB - We report of a case of bilateral limbus involvement by white lesions. A 72-year old man was referred for bilateral lesions involving the inferior part of the corneal limbus. White excrescences associated with a vascular pannus were observed at biomicroscope examination. Histologic patterns showed degenerative fibroelastosis. Degenerative fibroelastosis can be observed at the inferior corneal limbus. PMID- 18404131 TI - [Congenital retinal macrovessel: angiographic and optical coherence tomography views]. PMID- 18404132 TI - SUMO1 enhances 17-beta estradiol's effect on CRH promoter activation through estrogen receptors. AB - OBJECTIVES: Human corticotrophin-releasing hormone (CRH) plays a pivotal role in the stress response. Its expression is under the control of various steroid hormones, such as estrogens. The transcriptional activity of the estrogen receptor (ER) may be modified by small-ubiquitin related modifier (SUMO1). In the present study, we aim to reveal the role of SUMO1 in the regulation of ER mediated CRH promoter activation. METHODS: CHO-K1 cells were transfected with human ER and SUMO1 expressing plasmids together with the CRH promoter reporter gene. CRH mRNA was detected in BE(2)C cells by real time PCR. RESULTS: We found that estradiol could elevate CRH promoter activity to a much higher level in cells co-transfected with ER and SUMO1 than that with ER alone, and that the enhancement was blocked by the ER inhibitor, ICI182,780. Furthermore, the endogenous CRH mRNA expression was also increased when the BE(2)C cell were transfected with ERalpha and SUMO1 in contrast to the transfection with ERalpha alone. CONCLUSIONS: Our results indicate that SUMO1 participates in the modulation of ER-mediated CRH mRNA expression which may be important for the regulation of the stress response. PMID- 18404133 TI - Some ADHD polymorphisms (in genes DAT1, DRD2, DRD3, DBH, 5-HTT) in case-control study of 100 subjects 6-10 age. AB - BACKGROUND: Pharmacological approach is the most effective way of treatment of ADHD and its early application prevents from the progress of secondary disorders. The study on present neurotransmitter systems in pathology of ADHD can be helpful in selecting appropriate drug, since there are used various substances with different mechanisms of functioning in treatment of the hyperkinetic syndrome. METHOD: Within our study there were selected the genes of dopaminergic (DRD2, DRD3, DAT1), noradrenergic (DBH) and serotoninergic (5-HTT) systems. With the use of molecular-genetic methods based on association strategy "case-control" there were analysed genes including 11 polymorphisms. The presence of risk alleles was examined in comparison of the sample of 100 ADHD children to a control group of another 100 subjects, who were checked by child psychiatrists and examined with the Conners test in order to exclude eventual cases with ADHD symptoms. RESULTS: Our research suggests the association of some genes with ADHD. It could be concluded: 1) the risk of ADHD is significantly increased in the presence of one risk allele in genes DRD2 (O.R.=7,5), 5-HTT (O.R.=2,7) and DAT1 (O.R.=1,6). 2) The risk of ADHD is significantly increased at homozygotes for risk alleles in genes DRD2 (O.R.=54,8), 5-HTT (O.R.=6,7) and DAT1 (O.R.=6,6). For polymorphisms G444A and C1603T in DBH, which were detected by univariant analysis, haplotype analysis was performed and resulted in conclusion that: 3) the risk of ADHD is significantly increased in the presence of allele DBH +444A as well as in the presence of allele DBH +1603T (O.R.=15). PMID- 18404134 TI - Relationship of vaginal microflora to PROM, pPROM and the risk of early-onset neonatal sepsis. AB - BACKGROUND: Infections are among the most frequent causes of premature delivery and premature discharges of amniotic fluid. The vaginal ecosystem significantly contributes to the development of these conditions. Premature rupture of membranes (PROM) and preterm premature rupture of membranes (pPROM) are associated with an increased risk of intra-amniotic infection. The intra-amniotic infection negatively affects perinatal morbidity and mortality of newborns. OBJECTIVES: Finding of relationship of vaginal microflora to PROM, pPROM and the risk of early-onset neonatal sepsis. METHODS: A prospective study was implemented in 152 women with singleton gestations with PROM (n=52) and pPROM (n=47); the control group included 53 women with physiologic pregnancy and delivery at normal term without PROM. In all the women, aerobic cultivations from the vagina and cervix for Chlamydia trachomatis were provided before initiation of antibiotic treatment, the microbial picture of vagina was examined, and the cultivation examination of urine was carried out. The placenta was subjected to histopathologic examination. For the diagnosis of early-onset sepsis, we used concentrations of cytokines IL-6, IL-8, TNF-alpha, and the adhesion molecule, ICAM-1, from the venous umbilical blood taken immediately after delivery and cutting of the umbilical cord. Demonstrated early neonatal sepsis served as a further criterion. RESULTS: The most frequent bacteriologic findings throughout the group were coagulase-negative Staphylococci, Ureaplasma, Candida albicans, and Streptococcus viridans. Women with a diagnosis of urinary tract infection or diabetes mellitus were excluded from the study. We found no statistically significant relationship between a specific bacterial strain and PROM and pPROM. We found a statistically significant association between the risk for intra amniotic infection and the finding of S. viridans (p<0.001). There was also a statistically significant relationship between the microbiologic picture of the vagina VI and infection risk (p<0.002). CONCLUSIONS: Based on results of the present study, it is clear that the use of cultivation and microscopic findings in the vagina and cervix for the timely diagnosis of the risk of early-onset neonatal sepsis is restricted. PMID- 18404135 TI - Neurodevelopmental disorders, maternal Rh-negativity, and Rho(D) immune globulins: a multi-center assessment. AB - BACKGROUND: Many formulations of Thimerosal (49.55% mercury by weight)-containing Rho(D) immune globulins (TCRs) were routinely administered to Rh-negative mothers in the US prior to 2002. OBJECTIVES: It was hypothesized: (1) if prenatal Rho(D) immune globulin preparation exposure was a risk factor for neurodevelopmental disorders (NDs) then more children with NDs would have Rh-negative mothers compared to controls; and (2) if Thimerosal in the Rho(D)-immune globulin preparations was the ingredient associated with NDs, following the removal of Thimerosal from all manufactured Rho(D)-immune globulin preparations from 2002 in the US the frequency of maternal Rh-negativity among children with NDs should be similar to control populations. METHODS: Maternal Rh-negativity was assessed at two sites (Clinic A-Lynchburg, VA; Clinic B-Rockville and Baltimore, MD) among 298 Caucasian children with NDs and known Rh-status. As controls, maternal Rh negativity frequency was determined from 124 Caucasian children (born 1987-2001) without NDs at Clinic A, and the Rh-negativity frequency was determined from 1,021 Caucasian pregnant mothers that presented for prenatal genetic care at Clinic B (1980-1989). Additionally, 22 Caucasian patients with NDs born from 2002 onwards (Clinics A and B) were assessed for maternal Rh-negativity. RESULTS: There were significant and comparable increases in maternal Rh-negativity among children with NDs (Clinic: A=24.2%), autism spectrum disorders (Clinic: A=28.3%, B=25.3%), and attention-deficit-disorder/attention-deficit-hyperactivity-disorder (Clinic: A=26.3%) observed at both clinics in comparison to both control groups (Clinic: A=12.1%, B=13.9%) employed. Children with NDs born post-2001 had a maternal Rh-negativity frequency (13.6%) similar to controls. CONCLUSION: This study associates TCR exposure with some NDs in children. PMID- 18404136 TI - Lateralized brain and neuroendocrine dysregulation as response to traumatic stress. AB - Stressful life events cause a variety of conditions affecting cerebral and neuroendocrine functions. Repeated stressful events also may determine sensitization leading to an increase in responsiveness to stress stimuli. Recent findings suggest that cognitive and emotional dysregulation related to traumatic stress likely is linked to defective inhibitory functions that may also lead to temporo-limbic seizure-like activity, increased vulnerability to stressors, and dysregulated asymmetry in neural activity patterns that may influence interhemispheric dissociation. Together recent data show that dysregulation in the brain asymmetry and mental functioning may be caused by stress-related activation that can influence also the peripheral endocrine glands through the HPA axis and other pathways connecting the CNS and the target endocrine glands. PMID- 18404137 TI - Laparoscopic management of ectopic pregnancy. AB - OBJECTIVES: To assess the diagnostic process, laparoscopic treatment and postoperative period in patients hospitalized with a suspicion of ectopic pregnancy. MATERIALS AND METHODS: 211 women who underwent a typical laparoscopic surgery due to ectopic pregnancy, from 1993 through to 2006, were included in the study. The diagnosis was based on the patient's history, gynecological examination, transvaginal ultrasound findings and beta hCG concentration. RESULTS: The abnormal uterine bleeding was the most often reported symptom at admission (70.1%). The typical triad for ectopic pregnancy - amenorrhoea, abnormal bleeding and pain were present in only 55 patients (26%). The most frequent ultrasound findings were abnormal echogenic structures on the side of operated ectopic pregnancy (77.3%). 15.7% of women had no abnormalities described. The most frequent site of ectopic pregnancy implantation was the ampulla of the Fallopian tube (59.7%). Salpingectomy was performed in 53.5% of the cases, while salpingotomy in 35%. The postoperative period was generally uneventful in the majority of patients. There were only two cases that required a second surgery because of the worsening general clinical condition of the patient. CONCLUSIONS: The ectopic pregnancy is currently diagnosed quite early and therefore can be treated with the use of endoscopic tools in a scheduled operation time. Nowadays, only a small percentage of women still require a sudden open surgery intervention. PMID- 18404138 TI - Effects of bisphosphonates on lipid metabolism. AB - OBJECTIVES: Bisphosphonates are widely used for the treatment of metabolic bone disorders and their effects on lipid metabolism have also been investigated. Some studies reported that bisphosphonates have beneficial effects on serum cholesterol levels. In this study we aimed to assess the effects of bisphosphonates on lipid levels in hyperlipidemic patients who received bisphosphonates because of osteoporosis. METHODS: 49 female patients (age: 54.2+/ 7.2 years) with diagnosis of osteoporosis and hyperlipidemia were enrolled. Patients received alendronate 10 mg/day and they were followed up for 6 months. Pretreatment total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol, apolipoprotein A1 and apolipoprotein B levels were measured and compared with post-treatment levels. RESULTS: Pretreatment and post-treatment levels of total cholesterol were 255.2+/-34.3; 233.02+/-37.0 mg/dL, triglyceride levels were 153.0+/-57.3; 129.1+/-54.4 mg/dL, and LDL levels were 170.7+/-30.5; 160.0+/-34.2 mg/dL, respectively. Reductions in total cholesterol, triglyceride and LDL cholesterol levels were statistically significant; whereas differences in HDL cholesterol, apolipoprotein-A1 and apolipoprotein-B levels were not significant. CONCLUSIONS: Data from our study suggest that alendronate therapy may have beneficial effects on lipid metabolism. Thus, when hyperlipidemia is detected in patients receiving bisphosphonates, it is considered reasonable to follow the patient for a while before initiating antihyperlipidemic agent to prevent unnecessary use of drugs. PMID- 18404139 TI - Effects of acute L-carnitine intake on metabolic and blood lactate levels of elite badminton players. AB - OBJECTIVES: Purpose of this study is to research the effects of acute L-Carnitine intake on badminton players' metabolic and blood lactate values. DESIGN: A total of 16 Turkish national badminton players (8 male, 8 female) were voluntarily participated into study. MaxVO2, MET, energy consumption, HR (heart rate), VE (minute ventilation), R (respiratory exchange ratio), AT (anaerobic threshold), oxygen pulse and blood lactate (LA) of subjects were measured by Sensormedics VmaxST and Accutrend Lactate Analyzer. The participants were subjected to the test protocol twice before and after 2g of L-Carnitine intake. The data were evaluated by the use of SPSS 13.0 for Windows. RESULTS: No significant differences were found between 1st. (without L-Carnitine intake) and 2nd. (with L Carnitine intake) measurements of female participants as regards to all measured parameters. There was a significant difference in EMHR (exercise maximum heart rate) of males between two measurements (p<0.05). However the differences in other parameters were not significant. AT values of female subjects were not significant difference (p>0.05). Respiratory exchange ratio of males was significantly different at anaerobic threshold (p<0.05). CONCLUSION: Results of this study show that L-carnitine intake one hour prior to the exercise has no effect on the metabolic and blood lactate values of badminton players. PMID- 18404140 TI - Effect of intrauterine infection and perinatal risk factors on serum concentrations of insulin like growth factor (IGF-I) in full-term and preterm newborns. AB - OBJECTIVES: IGF-I is believed to be a key factor in fetal growth dynamics It is widely known, that serious early-onset infection in the newborn is a risk factor for further developmental disturbances in a child. However, effect of congenital infection as well as an influence of infectious and non-infectious perinatal risk factors on circulating IGF-I concentrations in newborns has not been examined, yet. DESIGN: Thus, the aim of this study was: 1) evaluation of IGF-I venous blood serum concentration in full-term and premature infants considering their sex, occurrence of intrauterine infection and perinatal risk factors; 2) establishing the relationship between IGF-I serum concentrations and chosen anthropometric parameters values in infected and healthy newborns. SETTING: The study involved 112 newborns appropriate for gestational age. Taking into consideration occurrence of early onset infection and gestational age we divided examined children into 4 groups: I group--infected, full-term newborns; II group--infected premature newborns; III group--healthy full-term newborns; IV group--healthy premature newborns. In all infants immediately after birth anthropometric measurements were performed (birth weight, body length, circumference of head and circumference of chest) and serum IGF-I concentration was determined. RESULTS: We demonstrated that full-term infants with intrauterine infection have statistically significantly higher concentration of IGF-I in blood serum than infected premature infants and healthy full-term infants. Analysis of correlation revealed a significant positive linear correlations between IGF-I serum concentration and gestational age and anthropometric parameters values. CONCLUSIONS: We conclude that intrauterine infection increases serum IGF-I concentration in full-term infants, but not in preterm infants, that may be a result of immaturity. We suggest serum IGF-I concentration may be considered an additional element of developmental and nutritional state assessment in infected newborn. PMID- 18404141 TI - Dihydrotestosterone and testosterone throughout the life span of Czech men. AB - OBJECTIVE: The dihydrotestosterone:testosterone ratio seems to be an important factor in the expression of androgenic activity, especially in the prostate and pilosebaceous unit. Whereas the decline of testosterone (T) in aging men is well known, controversial data can be found in literature concerning the age dependence of dihydrotestosterone (DHT) levels. Data from the database of the Institute of Endocrinology served as the basis for the definition of the life span curve for the ratio dihydrotestosterone : testosterone. METHODS: The results of testosterone and dihydrotestosterone obtained immunoassays from 10 251 male patients were used in the construction of the curve. RESULTS: The data show that after a peak of DHT:T in infancy and a subsequent decrease in puberty, the ratio of both androgens remains practically without change from approx. 20 years of age till old age. CONCLUSION: High DHT:T ratio in infancy decreases at puberty and throughout the entire reproductive period of life this ration remains practically constant. PMID- 18404142 TI - Depression, cortisol and somatoform dissociative symptoms. AB - OBJECTIVE: According to recent findings neuroendocrine response related to dissociative symptoms is related to dysregulation of the hypothalamus-pituitary adrenal (HPA) axis but HPA axis functioning as related to dissociation is only partially understood. METHOD: With the aim to test the relationship between basal serum cortisol and dissociative symptoms measured as somatoform and psychic dissociation we performed clinical testing and biochemical analysis in 30 inpatients with diagnosis of unipolar depression (mean age 41.46, SD=13.68). RESULTS: The results show that cortisol as an index of HPA axis functioning manifests significant relationship to somatoform dissociative symptoms (r=-0.40; p=0.014). CONCLUSIONS: The result indicates relationship between HPA-axis reactivity and somatoform dissociative symptoms in unipolar depressive patients and suggests that somatoform dissociation presents a defense mechanism related to a passive coping response. PMID- 18404144 TI - Clinical endocannabinoid deficiency (CECD): can this concept explain therapeutic benefits of cannabis in migraine, fibromyalgia, irritable bowel syndrome and other treatment-resistant conditions? AB - OBJECTIVES: This study examines the concept of clinical endocannabinoid deficiency (CECD), and the prospect that it could underlie the pathophysiology of migraine, fibromyalgia, irritable bowel syndrome, and other functional conditions alleviated by clinical cannabis. METHODS: Available literature was reviewed, and literature searches pursued via the National Library of Medicine database and other resources. RESULTS: Migraine has numerous relationships to endocannabinoid function. Anandamide (AEA) potentiates 5-HT1A and inhibits 5-HT2A receptors supporting therapeutic efficacy in acute and preventive migraine treatment. Cannabinoids also demonstrate dopamine-blocking and anti-inflammatory effects. AEA is tonically active in the periaqueductal gray matter, a migraine generator. THC modulates glutamatergic neurotransmission via NMDA receptors. Fibromyalgia is now conceived as a central sensitization state with secondary hyperalgesia. Cannabinoids have similarly demonstrated the ability to block spinal, peripheral and gastrointestinal mechanisms that promote pain in headache, fibromyalgia, IBS and related disorders. The past and potential clinical utility of cannabis-based medicines in their treatment is discussed, as are further suggestions for experimental investigation of CECD via CSF examination and neuro-imaging. CONCLUSION: Migraine, fibromyalgia, IBS and related conditions display common clinical, biochemical and pathophysiological patterns that suggest an underlying clinical endocannabinoid deficiency that may be suitably treated with cannabinoid medicines. PMID- 18404143 TI - Correlation between serum thyroxine and complements in patients with multiple sclerosis and neuromyelitis optica. AB - OBJECTIVE: The aim of this study is to assess serum levels of thyroid hormones (TH) and complement C3, C4, CH50 in patients with multiple sclerosis (MS) and neuromyelitis optica (NMO), and investigate whether there is a correlation between serum TH and complement factors in these patients. METHODS: One hundred and eighty-eight MS patients and sixty-five NMO patients were included in this study. The levels of serum TH were measured by magnetic antibody enzyme linking immunoassay and complement C3, C4 and CH50 were determined by immunoturbidimetry. RESULTS: The levels of TT4, complement C4 and CH50 in the serum were significantly higher in NMO patients than that in the MS. There were no significant differences in serum TT3, FT3, FT4 concentrations between NMO and MS patient. In MS patients, significant correlations between mean EDSS levels and serum C3 (r=0.223, p=0.014), and serum C4 (r=0.216, p=0.017) were found, and serum TT3 was positively correlated with serum CH50 (r=0.342, p=0.01). In NMO patients, the positive association between serum FT4 and C4 (r=0.533, p=0.006) were also found. CONCLUSIONS: These results indicate that NMO patients has different serum TH and complement C3, C4 and CH50 levels from MS patients, and that serum TH levels is correlated with CH50 and C4 in these patients, suggesting TH may play a different role in modulating the complement activation in MS and NMO. PMID- 18404145 TI - CD4+CD25high regulatory cells in peripheral blood of cancer patients. AB - AIM: Regulatory T cells (Treg) that prevent autoimmune diseases by suppression of self-reactive T cells may also suppress the immune response against cancer. Experimental tumor models in mice revealed that Tregs are potent inhibitors of an antitumor immune response. The purpose of the study was to identify a CD4+ population of regulatory T cells expressing high levels of CD25(CD4+CD25 high) in the peripheral blood of cancer patients and provide the opportunity to determine whether cancer patients exhibit an expanded CD4+CD25high pool. METHODS: The frequency of CD4+CD25high in the peripheral blood of 62 cancer patients and 15 healthy donors was determined by flow cytometry. RESULTS: Compared with healthy donors, cancer patients have an increasing prevalence of CD4+CD25high T cells in the peripheral blood with characteristics of Tregs, i.e. they are CD45-RA(), CD69(-). Among patients, those with higher percentages of CD4+CD25high T cells had a poor prognosis than did those with lower percentages. CONCLUSION: We provide evidence of an increased pool of CD4+CD25high in the peripheral blood of cancer patients, which may be related to immunosuppression and tumor progress in cancer patients. This finding suggests that the use of immunomodulatory therapy to treat cancer patients may be an effective strategy. PMID- 18404147 TI - Pseudogene-derived small interfering RNAs regulate gene expression in mouse oocytes. AB - Pseudogenes populate the mammalian genome as remnants of artefactual incorporation of coding messenger RNAs into transposon pathways. Here we show that a subset of pseudogenes generates endogenous small interfering RNAs (endo siRNAs) in mouse oocytes. These endo-siRNAs are often processed from double stranded RNAs formed by hybridization of spliced transcripts from protein-coding genes to antisense transcripts from homologous pseudogenes. An inverted repeat pseudogene can also generate abundant small RNAs directly. A second class of endo siRNAs may enforce repression of mobile genetic elements, acting together with Piwi-interacting RNAs. Loss of Dicer, a protein integral to small RNA production, increases expression of endo-siRNA targets, demonstrating their regulatory activity. Our findings indicate a function for pseudogenes in regulating gene expression by means of the RNA interference pathway and may, in part, explain the evolutionary pressure to conserve argonaute-mediated catalysis in mammals. PMID- 18404148 TI - Detection, clinical relevance and specific biological properties of disseminating tumour cells. AB - Most cancer deaths are caused by haematogenous metastatic spread and subsequent growth of tumour cells at distant organs. Disseminating tumour cells present in the peripheral blood and bone marrow can now be detected and characterized at the single-cell level. These cells are highly relevant to the study of the biology of early metastatic spread and provide a diagnostic source in patients with overt metastases. Here we review the evidence that disseminating tumour cells have a variety of uses for understanding tumour biology and improving cancer treatment. PMID- 18404149 TI - High-throughput kinase profiling as a platform for drug discovery. AB - To fully exploit the potential of kinases as drug targets, novel strategies for the efficient discovery of inhibitors are required. In contrast to the traditional, linear process of inhibitor discovery, high-throughput kinase profiling enables a parallel approach by interrogating compounds against hundreds of targets in a single screen. Compound potency and selectivity are determined simultaneously, providing a choice of targets to pursue that is guided by the quality of lead compounds available, rather than by target biology alone. PMID- 18404146 TI - Endogenous siRNAs from naturally formed dsRNAs regulate transcripts in mouse oocytes. AB - RNA interference (RNAi) is a mechanism by which double-stranded RNAs (dsRNAs) suppress specific transcripts in a sequence-dependent manner. dsRNAs are processed by Dicer to 21-24-nucleotide small interfering RNAs (siRNAs) and then incorporated into the argonaute (Ago) proteins. Gene regulation by endogenous siRNAs has been observed only in organisms possessing RNA-dependent RNA polymerase (RdRP). In mammals, where no RdRP activity has been found, biogenesis and function of endogenous siRNAs remain largely unknown. Here we show, using mouse oocytes, that endogenous siRNAs are derived from naturally occurring dsRNAs and have roles in the regulation of gene expression. By means of deep sequencing, we identify a large number of both approximately 25-27-nucleotide Piwi interacting RNAs (piRNAs) and approximately 21-nucleotide siRNAs corresponding to messenger RNAs or retrotransposons in growing oocytes. piRNAs are bound to Mili and have a role in the regulation of retrotransposons. siRNAs are exclusively mapped to retrotransposons or other genomic regions that produce transcripts capable of forming dsRNA structures. Inverted repeat structures, bidirectional transcription and antisense transcripts from various loci are sources of the dsRNAs. Some precursor transcripts of siRNAs are derived from expressed pseudogenes, indicating that one role of pseudogenes is to adjust the level of the founding source mRNA through RNAi. Loss of Dicer or Ago2 results in decreased levels of siRNAs and increased levels of retrotransposon and protein-coding transcripts complementary to the siRNAs. Thus, the RNAi pathway regulates both protein-coding transcripts and retrotransposons in mouse oocytes. Our results reveal a role for endogenous siRNAs in mammalian oocytes and show that organisms lacking RdRP activity can produce functional endogenous siRNAs from naturally occurring dsRNAs. PMID- 18404151 TI - Science amongst the vines. Meeting on signalling systems. PMID- 18404152 TI - To the centre of the volcano. Workshop on the mechanisms of nucleocytoplasmic transport. PMID- 18404150 TI - The TWEAK-Fn14 cytokine-receptor axis: discovery, biology and therapeutic targeting. AB - TWEAK is a multifunctional cytokine that controls many cellular activities including proliferation, migration, differentiation, apoptosis, angiogenesis and inflammation. TWEAK acts by binding to Fn14, a highly inducible cell-surface receptor that is linked to several intracellular signalling pathways, including the nuclear factor-kappaB (NF-kappaB) pathway. The TWEAK-Fn14 axis normally regulates various physiological processes, in particular it seems to play an important, beneficial role in tissue repair following acute injury. Furthermore, recent studies have indicated that TWEAK-Fn14 axis signalling may contribute to cancer, chronic autoimmune diseases and acute ischaemic stroke. This Review provides an overview of TWEAK-Fn14 axis biology and summarizes the available data supporting the proposal that both TWEAK and Fn14 should be considered as potential targets for the development of novel therapeutics. PMID- 18404153 TI - The histone-binding protein COPR5 is required for nuclear functions of the protein arginine methyltransferase PRMT5. AB - Protein arginine methyltransferase 5 (PRMT5) targets nuclear and cytoplasmic proteins. Here, we identified a nuclear protein, called cooperator of PRMT5 (COPR5), involved in the nuclear functions of PRMT5. COPR5 tightly binds to PRMT5, both in vitro and in living cells, but not to other members of the PRMT family. PRMT5 bound to COPR5 methylates histone H4 (R3) preferentially when compared with histone H3 (R8), suggesting that COPR5 modulates the substrate specificity of nuclear PRMT5-containing complexes, at least towards histones. Markedly, recombinant COPR5 binds to the amino terminus of histone H4 and is required to recruit PRMT5 to reconstituted nucleosomes in vitro. Consistently, COPR5 depletion in cells strongly reduces PRMT5 recruitment on chromatin at the PRMT5 target gene cyclin E1 (CCNE1) in vivo. Moreover, both COPR5 depletion and overexpression affect CCNE1 promoter expression. We propose that COPR5 is an important chromatin adaptor for PRMT5 to function on a subset of its target genes. PMID- 18404154 TI - Death through a tragedy: mitotic catastrophe. AB - Mitotic catastrophe (MC) has long been considered as a mode of cell death that results from premature or inappropriate entry of cells into mitosis and can be caused by chemical or physical stresses. Whereas it initially was depicted as the main form of cell death induced by ionizing radiation, it is today known to be triggered also by treatment with agents influencing the stability of microtubule, various anticancer drugs and mitotic failure caused by defective cell cycle checkpoints. Although various descriptions explaining MC exist, there is still no general accepted definition of this phenomenon. Here, we present evidences indicating that death-associated MC is not a separate mode of cell death, rather a process ('prestage') preceding cell death, which can occur through necrosis or apoptosis. The final outcome of MC depends on the molecular profile of the cell. PMID- 18404155 TI - Redox survival signalling in retina-derived 661W cells. AB - Reactive oxygen species have been implicated in processes involving cellular damage and subsequent cell death, especially in organs such as the eye that are constantly exposed to excitatory signals. However, recent studies have shown that oxidant species can also act as intracellular signalling molecules promoting cell survival, but little is known about this mechanism in the retina. The present study demonstrates for the first time that hydrogen peroxide (H2O2) is generated rapidly and acts as a pro-survival signal in response to a variety of apoptotic stimuli in retina-derived 661W cells and in the retinal ganglion cell line RGC-5. Focussing on 661Ws and serum deprivation, we systematically investigated pro survival and pro-death pathways and discovered that the rapid and transient burst of H2O2 activates the AKT survival pathway. Activation of the apoptotic machinery takes place following the decline of H2O2 to basal levels. To substantiate this proposed pro-survival role of H2O2, we inhibited the oxidant burst, which exacerbated cell death. Conversely, maintenance of the oxidant signal using exogenous H2O2 enhanced cell survival. Overall, the results presented in this study provide evidence for a novel role of H2O2 in mediating survival of retinal cells in response to apoptotic stimuli. PMID- 18404157 TI - Long live the cell death! PMID- 18404156 TI - BIM and other BCL-2 family proteins exhibit cross-species conservation of function between zebrafish and mammals. AB - Here we investigate the function of zebrafish Bcl-2 family proteins and demonstrate important conservation of function across zebrafish and mammalian systems. We have isolated a zebrafish ortholog of mammalian BIM and show that it is the most toxic of the zebrafish BH3-only genes examined, sharing this characteristic with the mammalian BIM gene. The zebrafish bad gene shows a complete lack of embryonic lethality, but like mammalian BAD, its pro-apoptotic activity is regulated through phosphorylation of critical serines. We also found that the pattern of mitochondrial dysfunction observed by zebrafish BH3 domain peptides in a mammalian cytochrome c release assay recapitulates the pattern of embryonic lethality induced by the respective mRNA injections in vivo. In contrast to zebrafish Bim, Bid exhibited only weak binding to zebrafish Bcl-2 and moderate-to-weak overall lethality in zebrafish embryos and isolated mitochondria. Given that zebrafish Bcl-2 binds strongly to mammalian BID and BIM peptides and proteins, the protein identified as the zebrafish Bid ortholog has different properties than mammalian BID. Overall, our results demonstrate the high degree of functional conservation between zebrafish and mammalian Bcl-2 family proteins, thus validating the zebrafish as a model system to further dissect the molecular mechanisms that regulate apoptosis in future forward genetic and chemical modifier screens. PMID- 18404158 TI - Oxygen-regulated protein-150 prevents calcium homeostasis deregulation and apoptosis induced by oxidized LDL in vascular cells. AB - Oxidized LDLs (oxLDLs) induce apoptosis, which contributes to the pathogenesis of atherosclerosis. The 150 kDa oxygen-regulated protein (ORP150), an endoplasmic reticulum (ER)-resident chaperone, is upregulated by hypoxia and prevents ischemia-induced cell death. The aim of this work was to investigate whether and how ORP150 can prevent apoptosis induced by oxLDLs in vascular cells. OxLDLs induced ORP150 expression in the ER of human microvascular endothelial cell line (HMEC-1). ORP150 expression was blocked by antioxidants, by the permeant calcium chelator BAPTA-AM, and by inhibitors of the inositol-1,4,5 trisphosphate (IP3) receptors, 2-aminoethyl diphenylborinate (2-APB) and xestospongin C. ORP150 silencing by siRNA-enhanced oxLDL-induced apoptosis, while forced ORP150 expression increased the resistance of cells via an inhibition of the oxLDL induced calcium rise, and of subsequent calpain activation, cytochrome c release, caspase 3 activation and apoptosis. A similar protective effect was achieved by BAPTA-AM, 2-APB and xestospongin C. Altogether, these data indicate that (i)ORP150 inhibits oxLDL-induced apoptosis by blocking calcium signaling and subsequent apoptosis, (ii)calcium released from ER stores through IP3 channels is involved in the oxLDL-induced calcium rise and apoptosis, and is inhibited by ORP150. Finally, ORP150 is expressed in advanced atherosclerotic lesions, where it may locally participate to reduce the apoptotic effect of oxLDLs and the subsequent risk of plaque rupture. PMID- 18404159 TI - Bevacizumab (Avastin) as a surgical adjunct in diabetic vitrectomy for fibrovascular disease. PMID- 18404160 TI - The application of human anterior lens capsule autotransplantation in phacotrabeculectomy: a prospective, comparative and randomized clinical study. AB - PURPOSE: To determine whether the autotransplantation of human anterior lens capsule (ALC) in the trabeculectomy site can aid filtration. METHODS: We conducted a prospective, randomized, and masked clinical trial to evaluate the outcome of human ALC autotransplantation in phacotrabeculectomy. Fifty patients with coexisting cataract and uncontrolled glaucoma despite maximum medical therapy were enrolled in this study. In the ALC group, combined phacoemulsification, posterior chamber intraocular lens implantation, and trabeculectomy were performed in 29 eyes, the anterior lens capsule was then put under the scleral tunnel. In the mitomycin-C (MMC) group, phacoemulsification, posterior chamber intraocular lens implantation, and trabeculectomy were performed in 21 eyes, MMC was used intraoperatively. Follow-up period was 12 months. The appearance of filtering bleb, intraocular pressure, and best corrected visual acuity were evaluated. RESULTS: (1) There was no statistical difference between the ALC group and the MMC group in the formation of functional filtering blebs (chi(2)=0.132, P>0.05), intraocular pressure (t=0.007, P>0.05), the number of antiglaucoma medications (Z=-0.800, P>0.05), and best corrected visual acuity (chi(2)=0.055, P>0.05). (2) The anterior lens capsule could be detected in the filtering site by ultrasound biomicroscopy 1 month after surgery. CONCLUSIONS: Anterior lens capsule autotransplantation could get comparable outcome as MMC in phacotrabeculectomy with few complications. PMID- 18404161 TI - Inadvertent administration of Olbas oil into the eye: a surprisingly frequent presentation. PMID- 18404162 TI - Floppy Iris Syndrome Hull Hooks (FISH Hooks): a new technique for managing IFIS in trabeculectomy surgery. PMID- 18404163 TI - Effects of DNAzymes targeting Aurora kinase A on the growth of human prostate cancer. AB - Aurora kinase A has been demonstrated to be involved in the malignant progression of many types of cancer including prostate cancer, we therefore hypothesized that Aurora kinase A might work as a valuable target for prostate cancer treatment. To test this hypothesis, we used DNAzyme technology to inhibit Aurora kinase A expression and evaluated the effects of DNAzymes as therapeutic agents to treat prostate cancer. In an in vitro cleavage assay, we found that a DNAzyme (DZ2) targeting Aurora kinase A could effectively cleave Aurora kinase A mRNA. When transfected into the prostate cancer cell line PC3, DZ2 was found to strongly inhibit the expression of Aurora kinase A examined by western blot analysis, and thus suppressed cell growth, arrested the progression of cell cycle, induced cell apoptosis and attenuated cell migration, as measured by 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltetrazolium assay, flow cytometry and Boyden chamber assay. Through in vivo study, we also found that DZ2 could significantly inhibit the growth of human prostate cancer xenografts in nude mice. In conclusion, DZ2 could effectively attenuate malignant progression of prostate cancer both in vitro and in vivo, suggesting that DNAzyme targeting Aurora kinase A may be used as a valuable therapy to treat prostate cancer. PMID- 18404165 TI - Influence of external cavity length on multimode hopping in microchip Nd:YAG lasers. AB - The influence of external cavity length on multimode hopping in microchip Nd:YAG lasers is investigated experimentally. With an optical feedback loop, the threshold gain of different longitudinal modes are all modulated by changing the external cavity length; a lambda/2 change in the external cavity length causes a one-period oscillation. The longitudinal modes can be divided into groups according to different initial threshold gain variations and modulation trends corresponding to different external cavity phases. Because of the initial gain difference, only one mode in each group is the dominant potential lasing mode, while others are suppressed. During the 2 pi change of the external cavity phase, mode hopping occurs among these potential lasing modes from different groups. Both the intensity waveforms and the number of hopping modes strongly depend on the external cavity length. Experimental results agree well with the theoretical analysis of the phenomenon of multimode hopping subjected to optical feedback in microchip Nd:YAG lasers. PMID- 18404166 TI - Measurement of steep aspheric surfaces using an anamorphic probe. AB - Synthetic aperture interferometry has been previously proposed as a possible in process method to measure aspheric form (R. Tomlinson, Appl. Opt.42, 701, 2003.). Preliminary demonstration utilized a scanning probe consisting of a pair of bare single mode fibers to perform source and receive functions. It was found that this probe did not have sufficient numerical aperture (NA) to measure steep surfaces and that simply increasing the NA decreases the light gathering efficiency substantially. In this paper, we introduce supplementary optics to increase the NA, and the light gathering efficiency has been increased by adopting an anamorphic design. A spherical test optic of known form is measured to demonstrate the capability of the new probe design. PMID- 18404167 TI - Control of the diffracted response of wire arrays with double periods. AB - The possibility of controlling the diffracted response of a periodic structure is investigated by using dual-period arrays, i.e., periodic arrays with a compound unit cell. We consider wire gratings in which each period comprises several cylinders with circular cross sections and all the cylinder axes are contained in the same plane. It is shown that this kind of structure permits one to control the diffracted response, regardless of the cylinder material and the incident polarization. Our numerical results suggest that the effect produced by wire gratings with dual-period characteristics is basically a geometric effect, and it can be present for other shapes of individual scatterers within each subarray. PMID- 18404168 TI - Smoothness-constrained projection method for particle analysis based on forward light scattering. AB - The iterative projection method, originally proposed by Kaczmarz and Cimmino, was recently applied to particle size analysis with forward light scattering. Modification was made to improve the convergent procedure. However, there are still limitations. It is found that the solutions are oscillatory when the method is applied to a set of underdetermined linear equations without any additional constraints. A new version of the projection method, combined with the constraints of nonnegativity and smoothness, is proposed and is studied in the presence of experimental noise. Compared with the early versions of the projection method, this one shows fast convergence, is more stable against random noise, and is insensitive to particle size distributions. PMID- 18404169 TI - Vertical surface emitting open coupled-cavities based on photonic crystal surface modes. AB - Two types of vertical surface emitting photonic crystal cavities based on beaming mechanism and coupled surface modes are studied. It is shown that vertical emission with a zero divergence angle and a high quality factor can be easily achieved by the back-to-back cavity design. The periodic modulation to the cavity surface alters nonradiative surface modes into radiative surface modes, and the constructive interference of the radiative waves gives rise to vertical emission and improves the quality of the output beam. A high quality factor can be attributed to the nonradiative surface mode on the cavity back whose small part of energy can be transferred into the cavity surface by coupling. The resonant property and the coupling efficiency of the cavities are investigated and optimal cavity configurations are obtained. These open coupled-cavities are good candidates of highly directional light sources. PMID- 18404170 TI - Estimation of optical properties of turbid media: experimental comparison of spatially and temporally resolved reflectance methods. AB - We compare two methods for the optical characterization of turbid media. The estimates of the absorption and reduced scattering coefficients (mu(a) and mu(')(s)) by a spatially resolved method and a time-resolved method are performed on tissue-like phantoms. Aqueous suspension of microspheres and Intralipid are used as scattering media with the addition of ink as an absorber. mu(')(s) is first measured on weakly absorbing media. The robustness of these measurements is then tested with respect to a variation of mu(a). The spatially resolved method gave more accurate estimates for mu(')(s) whereas the time-resolved method gave better results for mu(a) estimates. PMID- 18404171 TI - Theory and optimization of hollow ducts. AB - Hollow ducts compressing and homogenizing the emission of laser diode arrays are cheap, low weight alternatives to lens ducts. An analytic model is presented describing hollow ducts including internal reflection losses, which is useful in optimizing these devices, especially in determining the optimum length for maximum transmission efficiency. As an example, a gold coated duct is presented showing high transmission efficiencies of >90% comparable to lens ducts. PMID- 18404172 TI - Radiation from asymmetric laser-induced plasmas collected by a lens or optical fiber. AB - A method of calculating radiation spectra of an asymmetric (ellipsoidal) laser induced plasma plume is developed for two cases, when the radiation is collected by a lens and by an optical fiber. The lens receives the radiation coming from the entire plasma plume, while the view sight of an optical fiber is restricted to an acceptance cone so that only the radiation coming with an incident angle smaller than the cone angle is collected. The method incorporates the solution of the radiative transfer equation along the line of sight. An optimal number of lines is found to achieve the numerical convergence with a relative error <1%. Several practical simulations are carried out that include different placements and orientations of the lens and optic fiber. The effect of a motion of the center of the mass of the plasma plume on the radiation spectra is also investigated. PMID- 18404173 TI - Noise and aliases in off-axis and phase-shifting holography. AB - We have compared the respective efficiencies of off-axis and phase-shifting holography in terms of noise and aliases removal. The comparison is made by analyzing holograms of a USAF target backlit with laser illumination, recorded with a charge-coupled device camera. We show that it is essential to remove the local oscillator beam noise, especially at low illumination levels. PMID- 18404174 TI - Design and performance of a ZnSe tetra-prism for homogeneous substrate heating using a CO2 laser for pulsed laser deposition experiments. AB - We report on the design and performance of a ZnSe tetra-prism for homogeneous substrate heating using a continuous wave CO(2) laser beam in pulsed laser deposition experiments. We discuss here three potential designs for homogenizing prisms and use ray-tracing modeling to compare their operation to an alternative square-tapered beam-pipe design. A square-pyramidal tetra-prism design was found to be optimal and was subjected to modeling and experimental testing to determine the influence of interference and diffraction effects on the homogeneity of the resultant intensity profile produced at the substrate surface. A heat diffusion model has been used to compare the temperature distributions produced when using various different source intensity profiles. The modeling work has revealed the importance of substrate thickness as a thermal diffuser in producing a resultant homogeneous substrate temperature distribution. PMID- 18404175 TI - Observation of delay enhancement in an ion-doped crystal. AB - We report the investigation of the reduction of the group velocity propagation resulting from the steep change of the refractive index by the coherent population oscillation in an erbium ion-doped optical fiber. We study fully the influences of the ion density and the temperature on the fractional and time delay. We find that the fractional delay can be decreased at high or low temperature. Moreover, we conclude that the temperature can be used as a control parameter to reduce distortion. PMID- 18404176 TI - Reflective liquid crystal wavefront corrector used with tilt incidence. AB - To allow angular separation of the beam reflected off a liquid crystal wavefront corrector from the incident beam, it is convenient to introduce a small incident angle. This avoids using a beam splitter and the associated energy losses. The effect of the tilt incidence on the liquid crystal wavefront corrector was investigated in this paper. For a parallel aligned liquid crystal wavefront corrector, a simplified model was established and used to analyze the change of the phase modulation under the tilt incidence. The simulated results showed that the effect of the tilt incidence on the phase modulation can be ignored when the angle of tilt incidence is less than 6 degrees. The phase modulation related to the incident angle was measured and the changing trend was similar to the calculated results. The effect of the tilt incidence on the diffraction efficiency of the liquid crystal wavefront corrector was also discussed. The simulated results indicated that the reduction of the diffraction efficiency is less than 1% for incidence angles under 3 degrees. Last, a closed loop correction experiment was done with an incident angle of 1 degrees. After correction, the averaged peak to valley (PV) and root mean square (RMS) of the wavefront were down to 0.15 lambda and 0.03 lambda, respectively, and a resolvable image was acquired. PMID- 18404177 TI - Optical dispersion of radiation-grafted fluoro-polymer. AB - Controlling the index of refraction of polymers plays an important role in their advanced nonlinear optical and electro-optical applications as well as in nanophotonics and biophotonic technologies. A radiation-induced grafting of acrylic acid (AAc) onto poly(tetrafluoroethylene-co-perfluorovinyl ether) (PFA) copolymer films was carried out to synthesize graft copolymer films using gamma irradiation by the mutual method. The resulted films were characterized by Fourier transform infrared spectroscopy. The grafting process is associated with cross-linking through which a considerable change in the material refractive index is achieved. The linear refractive index, optical dispersion, and the quantum parameters of grafted poly(tetrafluoroethylene-co-perfluorovinyl ether) (PFA-g-PAAc) polymeric film are determined in a wide spectral range of 0.2-3 microm. The wavelength for zero material dispersion is evaluated. The oscillator, dispersion, and lattice energies, respectively, are calculated revealing the optical properties of the studied trunk polymeric substrate and the grafted ones. The origin of the optical properties has been discussed. PMID- 18404164 TI - The ERBB3 receptor in cancer and cancer gene therapy. AB - ERBB3, a member of the epidermal growth factor receptor (EGFR) family, is unique in that its tyrosine kinase domain is functionally defective. It is activated by neuregulins, by other ERBB and nonERBB receptors as well as by other kinases, and by novel mechanisms. Downstream it interacts prominently with the phosphoinositol 3-kinase/AKT survival/mitogenic pathway, but also with GRB, SHC, SRC, ABL, rasGAP, SYK and the transcription regulator EBP1. There are likely important but poorly understood roles for nuclear localization and for secreted isoforms. Studies of ERBB3 expression in primary cancers and of its mechanistic contributions in cultured cells have implicated it, with varying degrees of certainty, with causation or sustenance of cancers of the breast, ovary, prostate, certain brain cells, retina, melanocytes, colon, pancreas, stomach, oral cavity and lung. Recent results link high ERBB3 activity with escape from therapy targeting other ERBBs in lung and breast cancers. Thus a wide and centrally important role for ERBB3 in cancer is becoming increasingly apparent. Several approaches for targeting ERBB3 in cancers have been tested or proposed. Small inhibitory RNA (siRNA) to ERBB3 or AKT is showing promise as a therapeutic approach to treatment of lung adenocarcinoma. PMID- 18404178 TI - Polarization-splitting common-path interferometer based on a zero-twist liquid crystal display. AB - We present a compact optical polarization-splitting common-path interferometer based on a zero-twist liquid crystal display (LCD). The LCD is encoded with a diffraction grating pattern and illuminated with a polarization state with both horizontal and vertical components. The polarization component perpendicular to the director axis of the liquid crystal molecules is not affected by the LCD and forms the reference beam. However, the polarization component parallel to the director axis is diffracted at an angle determined by the period of the grating. By imposing an analyzer polarizer, these two beams create an interferogram that can either display retardance patterns encoded onto the LCD or analyze external birefringent optical elements. The programmability of the system allows new ways of increasing the utility of the interferograms. Experimental results are provided, including the visualization of optical vortices with different and opposite topological charges. PMID- 18404179 TI - Design of pitch conversion component for formation of multibeam optical recording head. AB - We describe a design of a planar lightwave circuit for parallel information processing using visible light. The circuit serves as a pitch conversion component (PCC) that can align multiple beams close together and easily composes a compact optical system that can project optical spots at a narrow pitch on a certain small plane. From the viewpoint of its application to optical recording, a PCC is designed to have over 50 waveguides according to the fabrication of waveguides for a blue-violet beam. It is analytically confirmed that a PCC contributes to the formation of a multibeam optical recording head with numerous beams. PMID- 18404180 TI - Detection of uranium in solids by using laser-induced breakdown spectroscopy combined with laser-induced fluorescence. AB - Detection of uranium in solids by using laser-induced breakdown spectroscopy has been investigated in combination with laser-induced fluorescence. An optical parametric oscillator wavelength-tunable laser was used to resonantly excite the uranium atoms and ions within the plasma plumes generated by a Q-switched Nd:YAG laser. Both atomic and ionic lines can be selected to detect their fluorescence lines. A uranium concentration of 462 ppm in a glass sample can be detected by using this technique at an excitation wavelength of 385.96 nm for resonant excitation of U II and a fluorescence line wavelength of 409.0 nm from U II. PMID- 18404181 TI - Doppler-free, multiwavelength acousto-optic deflector for two-photon addressing arrays of Rb atoms in a quantum information processor. AB - We demonstrate a dual wavelength acousto-optic deflector (AOD) designed to deflect two wavelengths to the same angles by driving with two RF frequencies. The AOD is designed as a beam scanner to address two-photon transitions in a two dimensional array of trapped neutral Rb87 atoms in a quantum computer. Momentum space is used to design AODs that have the same diffraction angles for two wavelengths (780 and 480 nm) and have nonoverlapping Bragg-matched frequency response at these wavelengths, so that there will be no cross talk when proportional frequencies are applied to diffract the two wavelengths. The appropriate crystal orientation, crystal shape, transducer size, and transducer height are determined for an AOD made with a tellurium dioxide crystal (TeO(2)). The designed and fabricated AOD has more than 100 resolvable spots, widely separated band shapes for the two wavelengths within an overall octave bandwidth, spatially overlapping diffraction angles for both wavelengths (780 and 480 nm), and a 4 micros or less access time. Cascaded AODs in which the first device upshifts and the second downshifts allow Doppler-free scanning as required for addressing the narrow atomic resonance without detuning. We experimentally show the diffraction-limited Doppler-free scanning performance and spatial resolution of the designed AOD. PMID- 18404182 TI - Generation of entangled photon pairs using small-coherence-time continuous wave pump lasers. AB - We address the generation of entangled photon pairs by parametric downconversion from solid state cw lasers with small coherence time. We consider a compact and low-cost setup based on a two-crystal scheme with type-I phase matching. We reconstruct the full density matrix by quantum tomography and analyze in detail the entanglement properties of the generated state as a function of the crystal's length and the coherence time of the pump. We verify the possibility to improve the visibility using a purification protocol based on a compensation crystal. PMID- 18404183 TI - Oxygen plasma flow properties deduced from laser-induced fluorescence and probe measurements. AB - Estimation of the local dissociation degree and the local mass-specific enthalpy of a pure oxygen plasma flow determined mainly from laser-induced fluorescence measurements are reported. Measurements have been conducted for several generator parameters in an inductively heated plasma wind tunnel. Additional probe measurements of total pressure together with the deduced translational temperature are used to estimate the local mass-specific enthalpy. For a reference condition, full dissociation has been measured. The measured translational temperature of atomic oxygen for this condition is T = 3500 K. Subsequently, the local mass-specific enthalpy has been derived using these local density and temperature measurements. For the reference condition the estimated value of h = 27 MJ/kg is in good agreement with the probe measurements and results from diode laser absorption spectroscopy. PMID- 18404184 TI - Ultraviolet femtosecond, picosecond and nanosecond laser microstructuring of silicon: structural and optical properties. AB - The effect of laser pulse duration on the morphology, composition, crystallinity and optical properties of self-organized Si microcones fabricated using 248 nm laser pulses (500 fs, 5 ps and 15 ns) in an SF(6) atmosphere, is presented in this paper. Despite distinct differences in the morphology, the Si cones show similar structure and composition independently of the laser pulse duration used: a core of single-crystalline Si, covered by a few hundred nanometer thick, sulfur doped nanocrystalline Si layer, where no amorphous Si is present. The obtained features exhibit strong below-bandgap absorptance, making them excellent candidates for Si based photodetectors with improved spectral response. PMID- 18404185 TI - Remote sensing of aerosol plumes: a semianalytical model. AB - A semianalytical model, named APOM (aerosol plume optical model) and predicting the radiative effects of aerosol plumes in the spectral range [0.4,2.5 microm], is presented in the case of nadir viewing. It is devoted to the analysis of plumes arising from single strong emission events (high optical depths) such as fires or industrial discharges. The scene is represented by a standard atmosphere (molecules and natural aerosols) on which a plume layer is added at the bottom. The estimated at-sensor reflectance depends on the atmosphere without plume, the solar zenith angle, the plume optical properties (optical depth, single scattering albedo, and asymmetry parameter), the ground reflectance, and the wavelength. Its mathematical expression as well as its numerical coefficients are derived from MODTRAN4 radiative transfer simulations. The DISORT option is used with 16 fluxes to provide a sufficiently accurate calculation of multiple scattering effects that are important for dense smokes. Model accuracy is assessed by using a set of simulations performed in the case of biomass burning and industrial plumes. APOM proves to be accurate and robust for solar zenith angles between 0 degrees and 60 degrees whatever the sensor altitude, the standard atmosphere, for plume phase functions defined from urban and rural models, and for plume locations that extend from the ground to a height below 3 km. The modeling errors in the at-sensor reflectance are on average below 0.002. They can reach values of 0.01 but correspond to low relative errors then (below 3% on average). This model can be used for forward modeling (quick simulations of multi/hyperspectral images and help in sensor design) as well as for the retrieval of the plume optical properties from remotely sensed images. PMID- 18404186 TI - Crystallography of ordered colloids using optical microscopy. 2. Divergent-beam technique. AB - A technique has been developed to extract quantitative crystallographic data from randomly oriented colloidal crystals using a divergent-beam approach. This technique was tested on a series of diverse experimental images of colloidal crystals formed from monodisperse suspensions of sterically stabilized poly (methyl methacrylate) spheres suspended in organic index-matching solvents. Complete sets of reciprocal lattice basis vectors were extracted in all but one case. When data extraction was successful, results appeared to be accurate to about 1% for lattice parameters and to within approximately 2 degrees for orientation. This approach is easier to implement than a previously developed parallel-beam approach with the drawback that the divergent-beam approach is not as robust in certain situations with random hexagonal close-packed crystals. The two techniques are therefore complimentary to each other, and between them it should be possible to extract quantitative crystallographic data with a conventional optical microscope from any closely index-matched colloidal crystal whose lattice parameters are compatible with visible wavelengths. PMID- 18404187 TI - Atmospheric turbulence profiling with SLODAR using multiple adaptive optics wavefront sensors. AB - The slope detection and ranging (SLODAR) method recovers atmospheric turbulence profiles from time averaged spatial cross correlations of wavefront slopes measured by Shack-Hartmann wavefront sensors. The Palomar multiple guide star unit (MGSU) was set up to test tomographic multiple guide star adaptive optics and provided an ideal test bed for SLODAR turbulence altitude profiling. We present the data reduction methods and SLODAR results from MGSU observations made in 2006. Wind profiling is also performed using delayed wavefront cross correlations along with SLODAR analysis. The wind profiling analysis is shown to improve the height resolution of the SLODAR method and in addition gives the wind velocities of the turbulent layers. PMID- 18404188 TI - Validation of a rotational coherent anti-Stokes Raman spectroscopy model for carbon dioxide using high-resolution detection in the temperature range 294-1143 K. AB - Experiments were performed in the temperature range of 294-1143 K in pure CO(2) using high-resolution rotational coherent anti-Stokes Raman spectroscopy (CARS), in the dual-broadband approach. Experimental single-shot spectra were recorded with high spectral resolution using a single-mode Nd:YAG laser and a relay imaging lens system on the exit of a 1 m spectrometer. A theoretical rotational CARS model for CO(2) was developed for evaluation of the experimental spectra. The evaluated mean temperatures of the recorded single-shot dual-broadband rotational coherent anti-Stokes Raman spectroscopy (DB-RCARS) spectra using this model showed good agreement with thermocouple temperatures, and the relative standard deviation of evaluated single-shot temperatures was generally 2-3%. Simultaneous thermometry and relative CO(2)/N(2)-concentration measurements were demonstrated in the product gas of premixed laminar CO/air flames at atmospheric pressure. Although the model proved to be accurate for thermometry up to 1143 K, limitations were observed at flame temperatures where temperatures were overestimated and relative CO(2)/N(2) concentrations were underestimated. Potential sources for these discrepancies are discussed. PMID- 18404189 TI - Biopolymer-based material used in optical image correlation. AB - We investigate the possible application of a modified deoxyribonucleic acid (DNA) dye system for dynamic processing of optical information, e.g., optical correlation. The system consists of a biopolymeric matrix made of DNA substituted with the cationic surfactant molecule cetyltrimethyl-ammonium chloride (CTMA) and doped with a photochromic Disperse Red 1 dye. Fast dynamics (millisecond range of rise and fall times) of output correlation signal formation was measured in a joint Fourier transform optical correlator experimental setup. Full reversibility of the correlation signal and reproducibility were observed even after long-time exposures. PMID- 18404190 TI - Fabrication and characterization of dye mixture doped polymer optical fiber as a broad wavelength optical amplifier. AB - Rhodamine 6G and Rhodamine B dye mixture doped polymer optical fiber amplifier (POFA), which can operate in a broad wavelength region (60 nm), has been successfully fabricated and tested. Tunable operation of the amplifier over a broad wavelength region is achieved by mixing different ratios of the dyes. The dye doped POFA is pumped axially using 532 nm, 10 ns laser pulses from a frequency doubled Q-switched Nd: YAG laser and the signals are taken from an optical parametric oscillator. A maximum gain of 22.3 dB at 617 nm wavelength has been obtained for a 7 cm long dye mixture doped POFA. The effects of pump energy and length of the fiber on the performance of the fiber amplifier are also studied. There exists an optimum length for which the amplifier gain is at a maximum value. PMID- 18404191 TI - Side illumination fluorescence emission characteristics from a dye doped polymer optical fiber under two-photon excitation. AB - Two-photon excited (TPE) side illumination fluorescence studies in a Rh6G-RhB dye mixture doped polymer optical fiber (POF) and the effect of energy transfer on the attenuation coefficient is reported. The dye doped POF is pumped sideways using 800 nm, 70 fs laser pulses from a Ti:sapphire laser, and the TPE fluorescence emission is collected from the end of the fiber for different propagation distances. The fluorescence intensity of RhB doped POF is enhanced in the presence of Rh6G as a result of energy transfer from Rh6G to RhB. Because of the reabsorption and reemission process in dye molecules, an effective energy transfer is observed from the shorter wavelength part of the fluorescence spectrum to the longer wavelength part as the propagation distance is increased in dye doped POF. An energy transfer coefficient is found to be higher at shorter propagation distances compared to longer distances. A TPE fluorescence signal is used to characterize the optical attenuation coefficient in dye doped POF. The attenuation coefficient decreases at longer propagation distances due to the reabsorption and reemission process taking place within the dye doped fiber as the propagation distance is increased. PMID- 18404192 TI - Detection of mechanical and disease stresses in citrus plants by fluorescence spectroscopy. AB - We have investigated the detection of mechanical and disease stresses in citrus plants (Citrus limonia [L.] Osbeck) using laser-induced fluorescence spectroscopy. Due to its economic importance we have chosen to investigate the citrus canker disease, which is caused by the Xanthomonas axonopodis pv. citri bacteria. Mechanical stress was also studied because it plays an important role in the plant's infection by such bacteria. A laser-induced fluorescence spectroscopy system, composed of a spectrometer and a 532 nm 10 mW excitation laser was used to perform fluorescence spectroscopy. The ratio of two chlorophyll fluorescence bands allows us to detect and discriminate between mechanical and disease stresses. This ability to discriminate may have an important application in the field to detect citrus canker infected trees. PMID- 18404193 TI - Dense range map reconstruction from a versatile robotic sensor system with an active trinocular vision and a passive binocular vision. AB - One major research issue associated with 3D perception by robotic systems is the creation of efficient sensor systems that can generate dense range maps reliably. A visual sensor system for robotic applications is developed that is inherently equipped with two types of sensor, an active trinocular vision and a passive stereo vision. Unlike in conventional active vision systems that use a large number of images with variations of projected patterns for dense range map acquisition or from conventional passive vision systems that work well on specific environments with sufficient feature information, a cooperative bidirectional sensor fusion method for this visual sensor system enables us to acquire a reliable dense range map using active and passive information simultaneously. The fusion algorithms are composed of two parts, one in which the passive stereo vision helps active vision and the other in which the active trinocular vision helps the passive one. The first part matches the laser patterns in stereo laser images with the help of intensity images; the second part utilizes an information fusion technique using the dynamic programming method in which image regions between laser patterns are matched pixel-by-pixel with help of the fusion results obtained in the first part. To determine how the proposed sensor system and fusion algorithms can work in real applications, the sensor system is implemented on a robotic system, and the proposed algorithms are applied. A series of experimental tests is performed for a variety of configurations of robot and environments. The performance of the sensor system is discussed in detail. PMID- 18404194 TI - Dual random phase encoding: a temporal approach for fiber optic applications. AB - We analyze the dual random phase encoding technique in the temporal domain to evaluate its potential application for secure data transmission in fiber optic links. To take into account the optical fiber multiplexing capabilities, the noise content of the signal is restricted when multiple channels are transmitted over a single fiber optic link. We also discuss some mechanisms for producing encoded time-limited as well as bandwidth-limited signals and a comparison with another recently proposed technique is made. Numerical simulations have been carried out to analyze the system performance. The results indicate that this multiplexing encryption method could be a good alternative compared with other well-established methods. PMID- 18404195 TI - Luminous power efficiency optimization of a white organic light-emitting diode by tuning its spectrum and its extraction efficiency. AB - We show an increase of the luminous power efficiency of a white organic light emitting diode (LED) with three emitters by optimizing its spectrum and its extraction efficiency. To calculate this efficiency we use a model with four parameters: the spectra, extraction efficiencies, internal quantum efficiencies of three emitters, and the driving voltage. This luminous power efficiency increases by 30% by use of a spectrum close to the spectrum of the MacAdam limit. This limit gives the highest luminous efficacy for a given chromaticity. We also show that a white organic LED with an inefficient deep blue emitter can give the same luminous power efficiency as a white organic LED with a more efficient light blue emitter, because of their different fractions in the radiant flux. Tuning the extraction efficiency with a microcavity to the spectrum also increases the luminous power efficiency by 10%. PMID- 18404201 TI - Phenotypic and genotypic characteristics of mastocytosis according to the age of onset. AB - Adult's mastocytosis is usually associated with persistent systemic involvement and c-kit 816 mutation, while pediatrics disease is mostly limited to the skin and often resolves spontaneously. We prospectively included 142 adult patients with histologically proven mastocytosis. We compared phenotypic and genotypic features of adults patients whose disease started during childhood (Group 1, n = 28) with those of patients whose disease started at adult's age (Group 2, n = 114). Genotypic analysis was performed on skin biopsy by sequencing of c-kit exons 17 and 8 to 13. According to WHO classification, the percentage of systemic disease was similar (75 vs. 73%) in 2 groups. C-kit 816 mutation was found in 42% and 77% of patients in groups 1 and 2, respectively (p<0.001). 816 c-kit mutation was associated with systemic mastocytosis in group 2 (87% of patients with systemic mastocytosis vs. 45% with cutaneous mastocytosis, p = 0.0001). Other c kit activating mutations were found in 23% of patients with mastocytosis' onset before the age of 5, 0% between 6 and 15 years and 2% at adults' age (p<0.001). In conclusion, pathogenesis of mastocytosis significantly differs according to the age of disease's onset. Our data may have major therapeutic relevance when considering c-kit-targeted therapy. PMID- 18404202 TI - Evaluation of paired-end sequencing strategies for detection of genome rearrangements in cancer. AB - Paired-end sequencing is emerging as a key technique for assessing genome rearrangements and structural variation on a genome-wide scale. This technique is particularly useful for detecting copy-neutral rearrangements, such as inversions and translocations, which are common in cancer and can produce novel fusion genes. We address the question of how much sequencing is required to detect rearrangement breakpoints and to localize them precisely using both theoretical models and simulation. We derive a formula for the probability that a fusion gene exists in a cancer genome given a collection of paired-end sequences from this genome. We use this formula to compute fusion gene probabilities in several breast cancer samples, and we find that we are able to accurately predict fusion genes in these samples with a relatively small number of fragments of large size. We further demonstrate how the ability to detect fusion genes depends on the distribution of gene lengths, and we evaluate how different parameters of a sequencing strategy impact breakpoint detection, breakpoint localization, and fusion gene detection, even in the presence of errors that suggest false rearrangements. These results will be useful in calibrating future cancer sequencing efforts, particularly large-scale studies of many cancer genomes that are enabled by next-generation sequencing technologies. PMID- 18404203 TI - The role of elastic stresses on leaf venation morphogenesis. AB - We explore the possible role of elastic mismatch between epidermis and mesophyll as a driving force for the development of leaf venation. The current prevalent 'canalization' hypothesis for the formation of veins claims that the transport of the hormone auxin out of the leaves triggers cell differentiation to form veins. Although there is evidence that auxin plays a fundamental role in vein formation, the simple canalization mechanism may not be enough to explain some features observed in the vascular system of leaves, in particular, the abundance of vein loops. We present a model based on the existence of mechanical instabilities that leads very naturally to hierarchical patterns with a large number of closed loops. When applied to the structure of high-order veins, the numerical results show the same qualitative features as actual venation patterns and, furthermore, have the same statistical properties. We argue that the agreement between actual and simulated patterns provides strong evidence for the role of mechanical effects on venation development. PMID- 18404204 TI - A generalized allosteric mechanism for cis-regulated cyclic nucleotide binding domains. AB - Cyclic nucleotides (cAMP and cGMP) regulate multiple intracellular processes and are thus of a great general interest for molecular and structural biologists. To study the allosteric mechanism of different cyclic nucleotide binding (CNB) domains, we compared cAMP-bound and cAMP-free structures (PKA, Epac, and two ionic channels) using a new bioinformatics method: local spatial pattern alignment. Our analysis highlights four major conserved structural motifs: 1) the phosphate binding cassette (PBC), which binds the cAMP ribose-phosphate, 2) the "hinge," a flexible helix, which contacts the PBC, 3) the beta(2,3) loop, which provides precise positioning of an invariant arginine from the PBC, and 4) a conserved structural element consisting of an N-terminal helix, an eight residue loop and the A-helix (N3A-motif). The PBC and the hinge were included in the previously reported allosteric model, whereas the definition of the beta(2,3) loop and the N3A-motif as conserved elements is novel. The N3A-motif is found in all cis-regulated CNB domains, and we present a model for an allosteric mechanism in these domains. Catabolite gene activator protein (CAP) represents a trans regulated CNB domain family: it does not contain the N3A-motif, and its long range allosteric interactions are substantially different from the cis-regulated CNB domains. PMID- 18404205 TI - Estimating cell depth from somatic mutations. AB - The depth of a cell of a multicellular organism is the number of cell divisions it underwent since the zygote, and knowing this basic cell property would help address fundamental problems in several areas of biology. At present, the depths of the vast majority of human and mouse cell types are unknown. Here, we show a method for estimating the depth of a cell by analyzing somatic mutations in its microsatellites, and provide to our knowledge for the first time reliable depth estimates for several cells types in mice. According to our estimates, the average depth of oocytes is 29, consistent with previous estimates. The average depth of B cells ranges from 34 to 79, linearly related to the mouse age, suggesting a rate of one cell division per day. In contrast, various types of adult stem cells underwent on average fewer cell divisions, supporting the notion that adult stem cells are relatively quiescent. Our method for depth estimation opens a window for revealing tissue turnover rates in animals, including humans, which has important implications for our knowledge of the body under physiological and pathological conditions. PMID- 18404206 TI - The emergence and fate of horizontally acquired genes in Escherichia coli. AB - Bacterial species, and even strains within species, can vary greatly in their gene contents and metabolic capabilities. We examine the evolution of this diversity by assessing the distribution and ancestry of each gene in 13 sequenced isolates of Escherichia coli and Shigella. We focus on the emergence and demise of two specific classes of genes, ORFans (genes with no homologs in present databases) and HOPs (genes with distant homologs), since these genes, in contrast to most conserved ancestral sequences, are known to be a major source of the novel features in each strain. We find that the rates of gain and loss of these genes vary greatly among strains as well as through time, and that ORFans and HOPs show very different behavior with respect to their emergence and demise. Although HOPs, which mostly represent gene acquisitions from other bacteria, originate more frequently, ORFans are much more likely to persist. This difference suggests that many adaptive traits are conferred by completely novel genes that do not originate in other bacterial genomes. With respect to the demise of these acquired genes, we find that strains of Shigella lose genes, both by disruption events and by complete removal, at accelerated rates. PMID- 18404207 TI - Binding-induced folding of a natively unstructured transcription factor. AB - Transcription factors are central components of the intracellular regulatory networks that control gene expression. An increasingly recognized phenomenon among human transcription factors is the formation of structure upon target binding. Here, we study the folding and binding of the pKID domain of CREB to the KIX domain of the co-activator CBP. Our simulations of a topology-based Go-type model predict a coupled folding and binding mechanism, and the existence of partially bound intermediates. From transition-path and Phi-value analyses, we find that the binding transition state resembles the unstructured state in solution, implying that CREB becomes structured only after committing to binding. A change of structure following binding is reminiscent of an induced-fit mechanism and contrasts with models in which binding occurs to pre-structured conformations that exist in the unbound state at equilibrium. Interestingly, increasing the amount of structure in the unbound pKID reduces the rate of binding, suggesting a "fly-casting"-like process. We find that the inclusion of attractive non-native interactions results in the formation of non-specific encounter complexes that enhance the on-rate of binding, but do not significantly change the binding mechanism. Our study helps explain how being unstructured can confer an advantage in protein target recognition. The simulations are in general agreement with the results of a recently reported nuclear magnetic resonance study, and aid in the interpretation of the experimental binding kinetics. PMID- 18404208 TI - Tentative T cells: memory cells are quick to respond, but slow to divide. AB - T cell memory is a cornerstone of protective immunity, and is the key element in successful vaccination. Upon encountering the relevant pathogen, memory T cells are thought to initiate cell division much more rapidly than their naive counterparts, and this is thought to confer a significant biological advantage upon an immune host. Here, we use traceable TCR-transgenic T cells to evaluate this proposed characteristic in CD4+ and CD8+ memory T cells. We find that, even in the presence of abundant antigen that was sufficient to induce in vivo IFNgamma production by memory T cells, both memory and naive T cells show an extended, and indistinguishable, delay in the onset of proliferation. Although memory cells can detect, and respond to, virus infection within a few hours, their proliferation did not begin until approximately 3 days after infection, and occurred simultaneously in all anatomical compartments. Thereafter, cell division was extraordinarily rapid for both naive and memory cells, with the latter showing a somewhat accelerated accumulation. We propose that, by permitting memory T cells to rapidly exert their effector functions while delaying the onset of their proliferation, evolution has provided a safeguard that balances the risk of infection against the consequences of severe T cell-mediated immunopathology. PMID- 18404209 TI - Human-like receptor specificity does not affect the neuraminidase-inhibitor susceptibility of H5N1 influenza viruses. AB - If highly pathogenic H5N1 influenza viruses acquire affinity for human rather than avian respiratory epithelium, will their susceptibility to neuraminidase (NA) inhibitors (the likely first line of defense against an influenza pandemic) change as well? Adequate pandemic preparedness requires that this question be answered. We generated and tested 31 recombinants of A/Vietnam/1203/04 (H5N1) influenza virus carrying single, double, or triple mutations located within or near the receptor binding site in the hemagglutinin (HA) glycoprotein that alter H5 HA binding affinity or specificity. To gain insight into how combinations of HA and NA mutations can affect the sensitivity of H5N1 virus to NA inhibitors, we also rescued viruses carrying the HA changes together with the H274Y NA substitution, which was reported to confer resistance to the NA inhibitor oseltamivir. Twenty viruses were genetically stable. The triple N158S/Q226L/N248D HA mutation (which eliminates a glycosylation site at position 158) caused a switch from avian to human receptor specificity. In cultures of differentiated human airway epithelial (NHBE) cells, which provide an ex vivo model that recapitulates the receptors in the human respiratory tract, none of the HA-mutant recombinants showed reduced susceptibility to antiviral drugs (oseltamivir or zanamivir). This finding was consistent with the results of NA enzyme inhibition assay, which appears to predict influenza virus susceptibility in vivo. Therefore, acquisition of human-like receptor specificity does not affect susceptibility to NA inhibitors. Sequence analysis of the NA gene alone, rather than analysis of both the NA and HA genes, and phenotypic assays in NHBE cells are likely to adequately identify drug-resistant H5N1 variants isolated from humans during an outbreak. PMID- 18404210 TI - Histoplasma requires SID1, a member of an iron-regulated siderophore gene cluster, for host colonization. AB - The macrophage is the primary host cell for the fungal pathogen Histoplasma capsulatum during mammalian infections, yet little is known about fungal genes required for intracellular replication in the host. Since the ability to scavenge iron from the host is important for the virulence of most pathogens, we investigated the role of iron acquisition in H. capsulatum pathogenesis. H. capsulatum acquires iron through the action of ferric reductases and the production of siderophores, but the genes responsible for these activities and their role in virulence have not been determined. We identified a discrete set of co-regulated genes whose transcription is induced under low iron conditions. These genes all appeared to be involved in the synthesis, secretion, and utilization of siderophores. Surprisingly, the majority of these transcriptionally co-regulated genes were found clustered adjacent to each other in the genome of the three sequenced strains of H. capsulatum, suggesting that their proximity might foster coordinate gene regulation. Additionally, we identified a consensus sequence in the promoters of all of these genes that may contribute to iron-regulated gene expression. The gene set included L-ornithine monooxygenase (SID1), the enzyme that catalyzes the first committed step in siderophore production in other fungi. Disruption of SID1 by allelic replacement resulted in poor growth under low iron conditions, as well as a loss of siderophore production. Strains deficient in SID1 showed a significant growth defect in murine bone-marrow-derived macrophages and attenuation in the mouse model of infection. These data indicated that H. capsulatum utilizes siderophores in addition to other iron acquisition mechanisms for optimal growth during infection. PMID- 18404211 TI - Molecular and cellular basis of microvascular perfusion deficits induced by Clostridium perfringens and Clostridium septicum. AB - Reduced tissue perfusion leading to tissue ischemia is a central component of the pathogenesis of myonecrosis caused by Clostridium perfringens. The C. perfringens alpha-toxin has been shown capable of inducing these changes, but its potential synergy with perfringolysin O (theta-toxin) is less well understood. Similarly, Clostridium septicum is a highly virulent causative agent of spontaneous gas gangrene, but its effect on the microcirculation has not been examined. Therefore, the aim of this study was to use intravital microscopy to examine the effects of C. perfringens and C. septicum on the functional microcirculation, coupled with the use of isogenic toxin mutants to elucidate the role of particular toxins in the resultant microvascular perfusion deficits. This study represents the first time this integrated approach has been used in the analysis of the pathological response to clostridial toxins. Culture supernatants from wild-type C. perfringens induced extensive cell death within 30 min, as assessed by in vivo uptake of propidium iodide. Furthermore, significant reductions in capillary perfusion were observed within 60 min. Depletion of either platelets or neutrophils reduced the alteration in perfusion, consistent with a role for these blood-borne cells in obstructing perfusion. In addition, mutation of either the alpha-toxin or perfringolysin O structural genes attenuated the reduction in perfusion, a process that was reversed by genetic complementation. C. septicum also induced a marked reduction in perfusion, with the degree of microvascular compromise correlating with the level of the C. septicum alpha-toxin. Together, these data indicate that as a result of its ability to produce alpha-toxin and perfringolysin O, C. perfringens rapidly induces irreversible cellular injury and a marked reduction in microvascular perfusion. Since C. septicum induces a similar reduction in microvascular perfusion, it is postulated that this function is central to the pathogenesis of clostridial myonecrosis, irrespective of the causative bacterium. PMID- 18404213 TI - An increase in mitochondrial DNA promotes nuclear DNA replication in yeast. AB - Coordination between cellular metabolism and DNA replication determines when cells initiate division. It has been assumed that metabolism only plays a permissive role in cell division. While blocking metabolism arrests cell division, it is not known whether an up-regulation of metabolic reactions accelerates cell cycle transitions. Here, we show that increasing the amount of mitochondrial DNA accelerates overall cell proliferation and promotes nuclear DNA replication, in a nutrient-dependent manner. The Sir2p NAD+-dependent de acetylase antagonizes this mitochondrial role. We found that cells with increased mitochondrial DNA have reduced Sir2p levels bound at origins of DNA replication in the nucleus, accompanied with increased levels of K9, K14-acetylated histone H3 at those origins. Our results demonstrate an active role of mitochondrial processes in the control of cell division. They also suggest that cellular metabolism may impact on chromatin modifications to regulate the activity of origins of DNA replication. PMID- 18404214 TI - Cell-to-cell stochastic variation in gene expression is a complex genetic trait. AB - The genetic control of common traits is rarely deterministic, with many genes contributing only to the chance of developing a given phenotype. This incomplete penetrance is poorly understood and is usually attributed to interactions between genes or interactions between genes and environmental conditions. Because many traits such as cancer can emerge from rare events happening in one or very few cells, we speculate an alternative and complementary possibility where some genotypes could facilitate these events by increasing stochastic cell-to-cell variations (or 'noise'). As a very first step towards investigating this possibility, we studied how natural genetic variation influences the level of noise in the expression of a single gene using the yeast S. cerevisiae as a model system. Reproducible differences in noise were observed between divergent genetic backgrounds. We found that noise was highly heritable and placed under a complex genetic control. Scanning the genome, we mapped three Quantitative Trait Loci (QTL) of noise, one locus being explained by an increase in noise when transcriptional elongation was impaired. Our results suggest that the level of stochasticity in particular molecular regulations may differ between multicellular individuals depending on their genotypic background. The complex genetic architecture of noise buffering couples genetic to non-genetic robustness and provides a molecular basis to the probabilistic nature of complex traits. PMID- 18404212 TI - Genomic islands in the pathogenic filamentous fungus Aspergillus fumigatus. AB - We present the genome sequences of a new clinical isolate of the important human pathogen, Aspergillus fumigatus, A1163, and two closely related but rarely pathogenic species, Neosartorya fischeri NRRL181 and Aspergillus clavatus NRRL1. Comparative genomic analysis of A1163 with the recently sequenced A. fumigatus isolate Af293 has identified core, variable and up to 2% unique genes in each genome. While the core genes are 99.8% identical at the nucleotide level, identity for variable genes can be as low 40%. The most divergent loci appear to contain heterokaryon incompatibility (het) genes associated with fungal programmed cell death such as developmental regulator rosA. Cross-species comparison has revealed that 8.5%, 13.5% and 12.6%, respectively, of A. fumigatus, N. fischeri and A. clavatus genes are species-specific. These genes are significantly smaller in size than core genes, contain fewer exons and exhibit a subtelomeric bias. Most of them cluster together in 13 chromosomal islands, which are enriched for pseudogenes, transposons and other repetitive elements. At least 20% of A. fumigatus-specific genes appear to be functional and involved in carbohydrate and chitin catabolism, transport, detoxification, secondary metabolism and other functions that may facilitate the adaptation to heterogeneous environments such as soil or a mammalian host. Contrary to what was suggested previously, their origin cannot be attributed to horizontal gene transfer (HGT), but instead is likely to involve duplication, diversification and differential gene loss (DDL). The role of duplication in the origin of lineage specific genes is further underlined by the discovery of genomic islands that seem to function as designated "gene dumps" and, perhaps, simultaneously, as "gene factories". PMID- 18404215 TI - Bmp4 is essential for the formation of the vestibular apparatus that detects angular head movements. AB - Angular head movements in vertebrates are detected by the three semicircular canals of the inner ear and their associated sensory tissues, the cristae. Bone morphogenetic protein 4 (Bmp4), a member of the Transforming growth factor family (TGF-beta), is conservatively expressed in the developing cristae in several species, including zebrafish, frog, chicken, and mouse. Using mouse models in which Bmp4 is conditionally deleted within the inner ear, as well as chicken models in which Bmp signaling is knocked down specifically in the cristae, we show that Bmp4 is essential for the formation of all three cristae and their associated canals. Our results indicate that Bmp4 does not mediate the formation of sensory hair and supporting cells within the cristae by directly regulating genes required for prosensory development in the inner ear such as Serrate1 (Jagged1 in mouse), Fgf10, and Sox2. Instead, Bmp4 most likely mediates crista formation by regulating Lmo4 and Msx1 in the sensory region and Gata3, p75Ngfr, and Lmo4 in the non-sensory region of the crista, the septum cruciatum. In the canals, Bmp2 and Dlx5 are regulated by Bmp4, either directly or indirectly. Mechanisms involved in the formation of sensory organs of the vertebrate inner ear are thought to be analogous to those regulating sensory bristle formation in Drosophila. Our results suggest that, in comparison to sensory bristles, crista formation within the inner ear requires an additional step of sensory and non sensory fate specification. PMID- 18404218 TI - Phase behavior of capillary bridges: towards nanoscale water phase diagram. AB - Water capillary bridges often condense at contact spots between small particles or asperities. The capillary adhesion force caused by these bridges is a major component of the attractive adhesion force, and thus it significantly affects the nanotribological performance of contacting surfaces. Recent atomic force microscope (AFM) measurements indicate that phase behavior of water in these tiny bridges may be different from macroscale water behavior. In particular, a metastable state with a deeply negative pressure, boiling at low temperatures, and ice at room temperature have been reported. Understanding these effects can lead to a modification of the traditional water phase diagram by creating a scale dependent or nanoscale phase diagram. PMID- 18404217 TI - A genome-wide gene expression signature of environmental geography in leukocytes of Moroccan Amazighs. AB - The different environments that humans experience are likely to impact physiology and disease susceptibility. In order to estimate the magnitude of the impact of environment on transcript abundance, we examined gene expression in peripheral blood leukocyte samples from 46 desert nomadic, mountain agrarian and coastal urban Moroccan Amazigh individuals. Despite great expression heterogeneity in humans, as much as one third of the leukocyte transcriptome was found to be associated with differences among regions. Genome-wide polymorphism analysis indicates that genetic differentiation in the total sample is limited and is unlikely to explain the expression divergence. Methylation profiling of 1,505 CpG sites suggests limited contribution of methylation to the observed differences in gene expression. Genetic network analysis further implies that specific aspects of immune function are strongly affected by regional factors and may influence susceptibility to respiratory and inflammatory disease. Our results show a strong genome-wide gene expression signature of regional population differences that presumably include lifestyle, geography, and biotic factors, implying that these can play at least as great a role as genetic divergence in modulating gene expression variation in humans. PMID- 18404216 TI - Beta-globin LCR and intron elements cooperate and direct spatial reorganization for gene therapy. AB - The Locus Control Region (LCR) requires intronic elements within beta-globin transgenes to direct high level expression at all ectopic integration sites. However, these essential intronic elements cannot be transmitted through retrovirus vectors and their deletion may compromise the therapeutic potential for gene therapy. Here, we systematically regenerate functional beta-globin intron 2 elements that rescue LCR activity directed by 5'HS3. Evaluation in transgenic mice demonstrates that an Oct-1 binding site and an enhancer in the intron cooperate to increase expression levels from LCR globin transgenes. Replacement of the intronic AT-rich region with the Igmu 3'MAR rescues LCR activity in single copy transgenic mice. Importantly, a combination of the Oct-1 site, Igmu 3'MAR and intronic enhancer in the BGT158 cassette directs more consistent levels of expression in transgenic mice. By introducing intron modified transgenes into the same genomic integration site in erythroid cells, we show that BGT158 has the greatest transcriptional induction. 3D DNA FISH establishes that induction stimulates this small 5'HS3 containing transgene and the endogenous locus to spatially reorganize towards more central locations in erythroid nuclei. Electron Spectroscopic Imaging (ESI) of chromatin fibers demonstrates that ultrastructural heterochromatin is primarily perinuclear and does not reorganize. Finally, we transmit intron-modified globin transgenes through insulated self-inactivating (SIN) lentivirus vectors into erythroid cells. We show efficient transfer and robust mRNA and protein expression by the BGT158 vector, and virus titer improvements mediated by the modified intron 2 in the presence of an LCR cassette composed of 5'HS2-4. Our results have important implications for the mechanism of LCR activity at ectopic integration sites. The modified transgenes are the first to transfer intronic elements that potentiate LCR activity and are designed to facilitate correction of hemoglobinopathies using single copy vectors. PMID- 18404219 TI - Oxidation and reactivity of nitrogen- and phosphorus-doped heterofullerenes. AB - Density functional theory (DFT) calculations suggest significantly different oxidation behaviour for phosphorus-doped heterofullerenes compared to their pure and nitrogen-doped counterparts, due to formation of a phosphene oxide. This oxide is not thermally labile, suggesting stable phosphofullerenes are likely to be C(59)POH and (C(59)PO)(2). In contrast, azafullerenes form stable epoxides when oxidised. We calculate the effect of oxidation on radical pairing and hydrogen passivation. Notably while the C(59)N radical behaves as a donor, C(59)PO will be an acceptor. PMID- 18404220 TI - Transport-limited current and microsonoreactor characterization at 3 low frequencies in the presence of water, acetonitrile and imidazolium-based ionic liquids. AB - A microsonoreactor, specially designed to carry out electrochemical tests in a room-temperature ionic liquid medium (RTIL), was studied. The cell, based on a particular design consisting of off-setting the ultrasonic probe out of the reaction volume, was characterized by several methods such as calorimetry, dosimetry and mass transfer measurements. The main result concerns the specific behaviour of the ionic liquid under ultrasonic irradiation. For example, the mass transfer enhancement is particularly high, characterized by an average Sherwood number of 6500 while the value obtained with an electrode rotating at 4500 rpm is only 1200. PMID- 18404221 TI - OMx-D: semiempirical methods with orthogonalization and dispersion corrections. Implementation and biochemical application. AB - The semiempirical methods of the OMx family (orthogonalization models OM1, OM2, and OM3) are known to describe biochemical systems more accurately than standard semiempirical approaches such as AM1. We investigate the benefits of augmenting these methods with an empirical dispersion term (OMx-D) taken from recent density functional work, without modifying the standard OMx parameters. Significant improvements are achieved for non-covalent interactions, with mean unsigned errors of 1.41 kcal/mol (OM2-D) and 1.31 kcal/mol (OM3-D) for the binding energy of the complexes in the JSCH-2005 data base. This supports the use of these augmented methods in quantum mechanical/molecular mechanical (QM/MM) studies of biomolecules, for example during system preparation and equilibration. As an illustrative application, we present QM and QM/MM calculations on the binding between antibody 34E4 and a hapten, where OM3-D performs better than the methods without dispersion terms (AM1, OM3). PMID- 18404222 TI - Amplified emission of phthalocyanine isolated in cryogenic matrices. AB - Laser induced fluorescence spectroscopy of free-base (H(2)Pc) and zinc (ZnPc) phthalocyanines trapped in rare gas and nitrogen matrices reveals a quite unexpected phenomenon with a moderate increase in the laser intensity. In all matrices except Xe, a huge increase occurs in the intensity of an emission band near 755 nm when pumping the S(1) <-- S(0) transition. The band involves a vibrational mode of the ground state, located at 1550 and 1525 cm(-1) for H(2)Pc and ZnPc, respectively. Many of the characteristics of amplified emission (AE) are exhibited by this vibronic transition. Excitation scans recorded for the AE band yield greatly enhanced site selectivity compared to what is obtained in normal fluorescence excitation scans. PMID- 18404223 TI - Methanol oxidation on a Pt(111)-OH/O surface. AB - The methanol oxidation on a hydroxylated Pt (Pt(111)-OH) surface has been investigated by means of infrared reflection absorption spectroscopy (IRAS) in ultra-high vacuum (UHV) and in acidic solution. The Pt(111)-OH surface in UHV was prepared by introducing water molecules on a Pt(111)-(2 x 2)-O surface and annealed at temperature higher than 160 K. Methanol was then, introduced to the Pt(111)-OH surface to show the dependence of the reaction intermediate on the annealing temperature. At an annealing temperature below 160 K, IR bands assignable to methanol overlayer were observed and no detectable intermediates, such as CO, formaldehyde and formate, were formed, suggesting that methanol molecules remain stable on Pt(111) surface without dissociation at this temperature region. At an annealing temperature above 160 K, on the other hand, CO and formate were observed. In addition, the oxidation of CO on Pt(111)-OH showed no sign of formate formation, indicating that formate is not derived from CO, but from a direct oxidation of methanol. Methanol oxidation was carried out in 0.1 mol dm(-3) HClO(4) solution on Pt(111) with a flow cell configuration and showed the formation of formate. These results indicate that the formate is the dominant non-CO intermediate both in UHV and in acidic solution, and the preadsorbed oxygen-containing species, in particular OH adsorbates, on Pt(111) surface plays a very important role in the formate formation process in methanol oxidation reaction. PMID- 18404224 TI - Solvent-dependent ultrafast internal conversion dynamics of n'-apo-beta carotenoic-n'-acids (n = 8, 10, 12). AB - The ultrafast internal conversion dynamics of 12'-apo-beta-carotenoic-12'-acid (12'CA), 10'-apo-beta-carotenoic-10'-acid (10'CA) and 8'-apo-beta-carotenoic-8' acid (8'CA) have been investigated by femtosecond pump-probe spectroscopy. The three apocarotenoic acids were excited to the S(2) state with different excess energies. Time constants tau(1) for the IC process S(1)/ICT --> S(0) were measured by probing the dynamics at 390 nm (S(0) --> S(2)), 575 nm (S(1)/ICT --> S(n)), 850, 860 and 890 nm (S(2) --> S(n) and S(1)/ICT --> S(0)). In nonpolar solvents, the observed reduction of the tau(1) values with increasing conjugation length of the acids is consistent with a reduction of the energy gap between the S(1)/ICT and S(0) states. The values are in good agreement with those of the corresponding apocarotenals studied previously in our groups. In polar solvents, a pronounced reduction of tau(1) values was observed for 12'CA, however the behavior was different from that observed for the respective aldehyde 12'-apo beta-caroten-12'-al studied previously: First, the degree of tau(1) reduction in methanol was milder for 12'CA (218 --> 55 ps) than for 12'-apo-beta-caroten-12' al (220 --> 8 ps). Secondly, for 12'CA the plateau of solvent independent tau(1) values extended further into the mid-polar range (up to 0.5 on the Deltaf scale) than previously observed for the 12'-aldehyde. For 10'CA the polarity effect on the tau(1) values was weaker ( approximately 71 ps in n-hexane and 34 ps in methanol) and for 8'CA it disappeared completely ( approximately 24 ps averaged over all solvents). The polarity-induced reduction of tau(1) is likely due to the stabilization of an intramolecular charge transfer state in polar solvents. This S(1)/ICT state is also responsible for the stimulated emission in the near IR, which has been observed in this specific class of carotenoids with a terminal carboxyl group for the first time. The occurrence of stimulated emission in the near IR region is also consistent with the steady-state fluorescence spectra which are reported along with the absorption spectra of these species. Possible reasons for the different behavior of the apocarotenoic acids compared to the respective aldehydes are discussed. PMID- 18404225 TI - On the electrodeposition of titanium in ionic liquids. AB - The ability to electrodeposit titanium at low temperatures would be an important breakthrough for making corrosion resistant layers on a variety of technically important materials. Ionic liquids have often been considered as suitable solvents for the electrodeposition of titanium. In the present paper we have extensively investigated whether titanium can be electrodeposited from its halides (TiCl(4), TiF(4), TiI(4)) in different ionic liquids, namely1-ethyl-3 methylimidazolium bis(trifluoromethylsulfonyl)amide ([EMIm]Tf(2)N), 1-butyl-1 methylpyrrolidinium bis(trifluoromethyl-sulfonyl)amide ([BMP]Tf(2)N), and trihexyltetradecyl-phosphonium bis(trifluoromethylsulfonyl)amide ([P(14,6,6,6)]Tf(2)N). Cyclic voltammetry and EQCM measurements show that, instead of elemental Ti, only non-stoichiometric halides are formed, for example with average stoichiometries of TiCl(0.2), TiCl(0.5) and TiCl(1.1). In situ STM measurements show that-in the best case-an ultrathin layer of Ti or TiCl(x) with thickness below 1 nm can be obtained. In addition, results from both electrochemical and chemical reduction experiments of TiCl(4) in a number of these ionic liquids support the formation of insoluble titanium cation-chloride complex species often involving the solvent. Solubility studies suggest that TiCl(3) and, particularly, TiCl(2) have very limited solubility in these Tf(2)N based ionic liquids. Therefore it does not appear possible to reduce Ti(4+) completely to the metal in the presence of chloride. Successful deposition processing for titanium in ionic liquids will require different maybe tailor-made titanium precursors that avoid these problems. PMID- 18404226 TI - Dissociative electron capture of halocarbon caused by the internal electron transfer from water trimer anion. AB - Dissociative electron capture dynamics of halocarbon absorbed on water cluster anion, caused by internal electron transfer from the water trimer anion to the halocarbon, have been investigated by means of the direct density functional theory (DFT)-molecular dynamics (MD) method. The CF(2)Cl(2) molecule and a water trimer anion e(-)(H(2)O)(3) were used as a halocarbon and a trapped electron, respectively. First, the structure of trapped electron state, expressed by e( )(H(2)O)(3)-CF(2)Cl(2), was fully optimized. The excess electron was trapped by a dipole moment of water trimer. Next, initial geometries were randomly generated around the equilibrium point of the trapped electron state, and then trajectories were run. The direct DFT-MD calculations showed that the spin density distribution of excess electron is gradually changed from the water cluster (trapped electron state) to CF(2)Cl(2) as a function of time. Immediately, the Cl(-) ion was dissociated from CF(2)Cl(2)(-) adsorbed on the water cluster. The reaction was schematically expressed by e(-)(H(2)O)(3)-CF(2)Cl(2)-->[(H(2)O)(3)- >-CF(2)CL(2)](-) --> (H(2O)(3) + CF(2)CL + CI(-) (I) where [(H(2)O)(3) CF(2)Cl(2)](-) indicates a transient intermediate state in which the excess electron is widely distributed on both the water cluster and CF(2)Cl(2). The mechanism of the electron capture of halocarbon from the trapped electron in water ice was discussed on the basis of the theoretical results. Also, the dynamics feature was compared with those of the direct electron capture reactions of CF(2)Cl(2) and CF(2)Cl(2)-(H(2)O)(3), i.e. e(-) + CF(2)Cl(2), and e(-) + CF(2)Cl(2)-(H(2)O)(3), investigated in our previous paper [Tachikawa and Abe, J. Chem. Phys., 2007, 126, 194310]. PMID- 18404227 TI - Influences of surfactant and nanoparticle assembly on effective interfacial tensions. AB - We have studied assembly at air-water and liquid-liquid interfaces with an emphasis on systems containing both surfactants and nanoparticles. Anionic surfactants, sodium dodecyl sulfate (SDS) and non-ionic surfactants, Triton X-100 and tetraethylene glycol alkyl ethers (C(8)E(4), C(12)E(4) and C(14)E(4)), effectively decrease the surface tension of air-water interfaces. The inclusion of negatively charged hydrophilic silica nanoparticles (diameters of approximately 13 nm) increases the efficiency of the SDS molecules but does not alter the performance of the non-ionic surfactants. The former is likely due to the repulsive Coulomb interactions between the SDS molecules and nanoparticles which promote the surfactant adsorption at air-water interfaces. For systems involving trichloroethylene (TCE)-water interfaces, the SDS and Triton X-100 surfactants effectively decrease the interfacial tensions and the nanoparticle effects are similar compared to those involving air-water interfaces. Interestingly, the C(12)E(4) and C(14)E(4) molecules, with or without the presence of nanoparticles, fail to decrease the TCE-water interfacial tensions. Our molecular dynamics simulations have suggested that the tetraethylene glycol alkyl ether molecules tend to disperse in the TCE phase rather than adsorb at the TCE-water interfaces. PMID- 18404228 TI - Photopolymerization of diacetylene-capped gold nanoparticles. AB - Gold nanoparticles (AuNPs) coated with the diacetylene henicosa-10,12-diyn-1-yl (DS9) disulfide were successfully prepared by direct synthesis in toluene solutions. The average size of the nanohybrid metal core was finely adjusted by manipulation of the preparative conditions in the diameter range from 1.6 to 7.5 nm, as determined by TEM characterization. The topochemical polymerization of DS9 chemisorbed onto the gold nanoclusters of different size was carried out in colloidal suspensions by exposure to UV radiation and the process was monitored by UV-Vis and Raman spectroscopies. The results showed that in these assemblies the monomer undergoes an intra-particle polymerization and that the dominant polydiacetylene phase present is ruled by the core size. The deposition of the photoirradiated colloids onto different substrates was found to leave the polydiacetylene conjugation unaltered. PMID- 18404229 TI - Control of the optical properties of CdTe nanocrystals by selective exchange of Te with thiolate: effect of organic ligands on the formation of core-shell structures. AB - Changes in the optical properties of CdTe nanocrystals through selective surface exchange reaction with thiolate molecules in the organic phase are studied with an aim to investigate the mechanism and the role of organic ligands. The reaction was mediated by dissociation of Te anions via oxidation in air from CdTe nanocrystals, followed by attachment of thiolate molecules in a 1:1 stoichiometric manner. This results in a gradual shell formation and a corresponding decrease in the size of the fluorescent CdTe cores, which induces a blue shift of both the absorption edge and emission wavelength in the visible region. A systematic study including the addition of ligands at different concentrations revealed that Te dissociation is the rate-determining step for the process and the degree of blue shift is significantly dependent on the amount of organic ligands present. The process could also be kinetically controlled through the addition of an excess amount of thiolate ligands, allowing systematic tuning of the emission properties of nanocrystals under ambient conditions. PMID- 18404230 TI - The interaction of water with Ni(111) and H/Ni(111) studied by TPD and HREELS. AB - We have used temperature-programmed desorption in combination with specular and off-specular high resolution electron energy loss spectroscopy to study the interaction of H(2)O and D(2)O with the bare and hydrogen-covered Ni(111) surface. Our results for the bare metal surface agree with previous reports and we are able to relate two prominent features in vibrational spectra to nuclear motions at the surface. Pre-covering Ni(111) with hydrogen alters both adsorption and desorption of water significantly. The strong H-Ni bond does not allow for isotopic exchange with co-adsorbed D(2)O. Strong resemblance of desorption traces and vibrational spectra of submonolayer coverages on H-covered Ni(111) and multilayers on bare Ni(111) suggests that adsorption of hydrogen makes this nickel surface hydrophobic. PMID- 18404231 TI - SERS-activating effect of chlorides on borate-stabilized silver nanoparticles: formation of new reduced adsorption sites and induced nanoparticle fusion. AB - Changes in morphology, surface reactivity and surface-enhancement of Raman scattering induced by modification of borate-stabilized Ag nanoparticles by adsorbed chlorides have been explored using TEM, EDX analysis and SERS spectra of probing adsorbate 2,2'-bipyridine (bpy) excited at 514.5 nm and evaluated by factor analysis. At fractional coverages of the parent Ag nanoparticles by adsorbed chlorides <0.6, the Ag colloid/Cl(-)/bpy systems were found to be constituted by fractal aggregates of Ag nanoparticles fairly uniform in size (10 +/- 2 nm) and SERS spectra of Ag(+)-bpy surface species were detected. The latter result was interpreted in terms of the presence of oxidized Ag(+) and/or Ag(n)(+) adsorption sites, which have been encountered also in systems with the chemically untreated Ag nanoparticles. At chloride coverages >0.6, a fusion of fractal aggregates into the compact aggregates of touching and/or interpenetrating Ag nanoparticles has been observed and found to be accompanied by the formation of another surface species, Ag-bpy, as well as by the increase of the overall SERS enhancement of bpy by factor of 40. The same Ag-bpy surface species has been detected under the strongly reducing conditions of reduction of silver nitrate by sodium borohydride in the presence of bpy. The formation of Ag-bpy is thus interpreted in terms of the stabilization of reduced Ag(0) adsorption sites by adsorbed bpy. The formation of reduced adsorption sites on Ag nanoparticle surfaces at chloride coverages >0.6 is discussed in terms of local changes in the work function of Ag. Finally, the SERS spectral detection of Ag-bpy species is proposed as a tool for probing the presence of reduced Ag(0) adsorption sites in systems with chemically modified Ag nanoparticles. PMID- 18404232 TI - Tuning the dynamics and molecular distribution of the self-spreading lipid bilayer. AB - The self-spreading dynamics of lipid bilayers were investigated at controlled electrolyte concentrations. The self-spreading velocity increased when the concentration of NaCl was increased from 1 to 100 mM. Comparing the experimentally determined spreading energy with that estimated from theoretical models, we found that the self-spreading dynamics were well explained by considering the van der Waals interaction, double layer interaction and hydration interaction energies between the self-spreading bilayer and the substrate. The characteristic behavior at high concentration is attributable to the increase in the density of the lipid layer, originating from the effective shielding of the molecular charges by the electrolyte ions in solution. The distribution of doped dye-labeled molecule within the spreading bilayer was also controllable by tuning the electrolyte concentration. All of these findings were explained by systematic changes in bilayer-substrate or inter-molecular interactions depending on the electrolyte concentration. PMID- 18404233 TI - Is the mobility of the pore walls and water molecules in the selectivity filter of KcsA channel functionally important? AB - We performed in-depth analysis of the forces which act on the K(+) ions in the selectivity filter of the KcsA channel in order to estimate the relative importance of static and dynamic influence of the filter wall and water molecules on ion permeation and selectivity. The forces were computed using the trajectories of all-atom molecular dynamics simulations. It is shown that the dynamics of the selectivity filter contributes about 3% to the net force acting on the ions and can be neglected in the studies focused on the macroscopic properties of the channel, such as the current. Among the filter atoms, only the pore-forming carbonyl groups can be considered as dynamic in the studies of microscopic events of conduction, while the dynamic effects from all other atoms are negligible. We also show that the dynamics of the water molecules in the filter can not be neglected. The fluctuating forces from the water molecules can be as strong as net forces from the pore walls and can effectively drive the ions through the local energy barriers in the filter. PMID- 18404234 TI - Effect of the electronic structure of carbon nanotubes on the selectivity of electrochemical functionalization. AB - The functionalization of carbon nanotubes through electrochemical routes is gaining importance due to the high degree of control achievable and the ability to render the tubes with a variety of chemical and biological species. In this article, we report systematic investigations on the grafting of phenyl groups through diazonium coupling onto individual metallic and semiconducting carbon nanotubes both experimentally and theoretically. The results show clearly that by optimizing the electrochemical conditions it is possible to obtain a high degree of selectivity for the coupling of phenyl radicals onto metallic nanotubes. The outlined conclusions have strong implications for the design of strategies for the controlled functionalization of individual single-wall carbon nanotubes. PMID- 18404235 TI - Effects of adsorbed water on plasmon-based dissolution, redeposition and resulting spectral changes of Ag nanoparticles on single-crystalline TiO2. AB - Ag nanoparticles photocatalytically deposited on single-crystalline TiO(2)(111) and (100) surfaces exhibit visible light-induced topographic and spectral changes corresponding to the light wavelength. A certain amount of adsorbed water on the surface is essential for the changes. The photoinduced behaviors strongly depend on the amount of the adsorbed water and on interparticle spacing, indicating that ionic conductivity on the TiO(2) surface is a key factor. The results obtained here are rationally explained in terms of a photoelectrochemical mechanism in which photoanodic dissolution of Ag nanoparticles to Ag(+) and cathodic re deposition of Ag play essential roles. PMID- 18404236 TI - Inter-regional variations in men's attitudes, suicide rates and sociodemographics in Quebec (Canada). AB - BACKGROUND: Suicide rates can vary quite considerably and attitudes regarding suicide may explain part of the variation. METHOD: The present study investigated, across the 17 regions of Quebec (Canada), men's attitudes towards suicide, expressing pain and help seeking, as they are related to suicide rates and sociodemographic variables (unemployment, divorce/separation, income and education). RESULTS: Most correlations were non-significant. However, in regions with an above-average educational level and with higher divorce/separation rates, men had better attitudes towards expressing pain. Furthermore, in regions where men were more inclined to express pain, suicide rates were lower. Also, significant positive correlations were found between suicide rates and low educational level, but also between an increase in suicide rates and an increase in income level. CONCLUSIONS: Where the aetiology of suicide is concerned, researchers must examine both sociodemographic factors and the psychological factors associated with them. PMID- 18404237 TI - Predicting detrusor overactivity using a physician-based scoring system. AB - Women with lower urinary tract symptoms (LUTS) commonly present to primary care. Management is often based on a symptomatic diagnosis alone although LUTS correlate with urodynamic diagnosis in 65% of cases. The aim of this study was to develop a scoring system to discriminate between the symptoms of stress urinary incontinence (SUI) and overactive bladder (OAB). Physicians completed a specially developed questionnaire for all women complaining of LUTS. Women then underwent videocystourethrography. A scoring system based on the likelihood of each symptom for DO was developed using odds ratios and 95% confidence intervals (CI). This scoring system was subsequently used to discriminate those women with OAB from those with SUI. One hundred seventy one women were recruited to the study. This scoring system had a sensitivity of 79%, a specificity of 78% and a positive predictive value of 73% to identify detrusor overactivity (DO) This new OAB scoring system may be used in primary care to help discriminate between USI and DO. PMID- 18404238 TI - Reply: "Treatment-of-choice for Buerger's disease (thromboangiitis obliterans): still an unresolved issue". PMID- 18404239 TI - An increased prevalence of fibromyalgia in iron deficiency anemia and thalassemia minor and associated factors. AB - In this study, we evaluated the prevalence of fibromyalgia (FM) in iron deficiency anemia (IDA) and thalassemia minor (TM) patients and associated factors. In addition, we investigated the prevalence of IDA in outpatients with fibromyalgia, and its effect on clinical findings. The study included 205 IDA, 40 TM patients and 100 healthy controls. FM was diagnosed according to 1990 ACR criteria. Whole blood count, biochemical tests, and serum iron parameters were determined. Pain, fatigue, and FM Impact Questionnaire (FIQ) functional item scores were assessed in FM subjects. In addition, the prevalence of IDA in FM patients diagnosed at the Rheumatology Outpatient Clinic was determined. The prevalences of FM in IDA (17.6%) and TM (20%) groups were higher than in controls (6%; p values 0.006 and 0.025, respectively). When IDA patients with FM were compared to those without FM, it was seen that a higher percentage were females, married, and a higher percentage had history of pica (all p values < 0.05). Serum hemoglobin and iron parameters did not differ between IDA patients with and without FM. IDA was detected in 48 (24.5%) of 196 FM patients. FM patients without IDA had higher sleep disturbance scores (p = 0.012) and longer duration of FM (p = 0.045). FM was a common finding in patients with IDA and TM. FM was associated with female sex and history of pica in IDA patients, and not associated with serum hemoglobin and selected iron parameters. The presence of FM in TM had no association with any of the above-mentioned parameters. PMID- 18404241 TI - Relationship of total body fat mass to bone area in New Zealand five-year-olds. AB - Fat mass was recently shown to be a positive determinant of bone mass and size independently of lean mass in a birth cohort of British 9-year-olds. The present study was undertaken to investigate whether similar relationships are evident in younger, preschool children. Height and weight were measured, and a total-body dual-energy X-ray absorptiometric scan was performed on 194 preschool New Zealand children (81 girls, 113 boys) participating in the Dunedin birth cohort Family, Lifestyle, Activity, Movement, and Eating (FLAME) study close to their fifth birthday. Relationships of total-body fat mass and lean mass to total-body-less head (TBLH) bone area and TBLH bone mineral content (BMC) were evaluated using linear regression. Girls had higher mean fat mass (3.9 vs. 3.2 kg) and lower lean mass (14.5 vs. 15.2 kg) than boys (P < 0.001), but their heights, weights, and TBLH bone area were similar. Although a given weight of lean tissue was associated with greater increases in TBLH area than a given weight of fat tissue, our results show that fat mass was an independent predictor of TBLH bone area (R (2 )= 0.79, P < 0.001) and TBLH BMC (R (2) = 0.74, P < 0.001) in data adjusted for socioeconomic status, ethnic group, lean mass, and height. We conclude that increased fat mass is associated with outward expansion of the TBLH skeletal envelope (wider bones) independently of height and lean mass in very young children. PMID- 18404242 TI - Abstracts of the 35th European Symposium on Calcified Tissues, 24-28 May 2008, Barcelona, Spain. PMID- 18404243 TI - Utility values associated with osteoporotic fracture: a systematic review of the literature. AB - We reviewed studies that have estimated the impact of osteoporotic fracture on quality-adjusted life years (QALY) and to determine reference values for countries that would like to carry out cost-utility analyses but that do not have their own values. The computerized medical literature databases Medline and EMBASE were searched from January 1990 to December 2006. The search was carried out in two steps. The first step was to identify studies that related to quality of life in osteoporosis. As part of the second step, only the studies that translated quality of life into a utility value (one single value for health status ranging 0-1) and calculated a utility loss over a period of at least 1 year were selected. From the 152 studies identified in the first analysis, only 16 were retained after the second step. Ten studies investigated utility values for hip fractures, 11 for vertebral fractures, five for distal forearm fractures, and four for other osteoporotic fractures and fracture interactions. Utility values differed substantially between studies, partly due to the valuation technique used, the severity of fractures, and the sample size. This review suggests that there is no meaningful average value across different studies, different samples, different countries, or different instruments. Although we tried to determine the best available values, these values do not preclude the need for country-specific studies. Finally, we also make recommendations regarding the design and methodology for such studies. PMID- 18404244 TI - Arterial anatomical basis of the dorsal digito-metacarpal flap for long fingers. AB - Several flaps have been described to treat severe soft tissue defects of the finger dorsal side. Many authors studied vascular organization of the hand on its dorsal side; most of them insisted on deep vascularization into the intermetacarpal spaces, which is formed by the dorsal metacarpal arteries. Those dorsal metacarpal arteries are the anatomical support of many flaps, which do not preserve the dorsal interosseous muscles fascias. Only few authors described dorsal vascular organization at the level of the proximal phalanx; however, using a rotation point of a flap distally to the metacarpal head with a donor site on the dorsal aspect of the hand could cover all distal soft tissue defect of long finger. In order to determine the technical limitations of dorsal digito metacarpal flap procedures, we studied number and location of arterial anastomoses between the reticular subcutaneous dorsal network and the rest of the vascularization at this level, which was formed by the deeper dorsal metacarpal arteries, common palmar digital arteries and proper palmar digital arteries, and between the dorsal digital arteries. Twenty-four long fingers from embalmed cadavers were studied after a reverse flow injection of colored latex and dissected layer-by-layer preserving the digital-metacarpal arterial network. At the level of the hand, the dorsal metacarpal arteries of the third and fourth intermetacarpal spaces were inconstant. When present, two or three arteries anastomosed in star shape with the reticular network. No such arterial anastomosis was observed proximally to the level of the intertendinous connections (junctura tendinorum) that bridge the extensor digitorum communis tendons. When no dorsal metacarpal artery was present, some communicant arteries arose from the common palmar digital arteries. Moreover, all the nutrient branches were more numerous distally to the intertendinous connections (junctura tendinorum). At the level of the metacarpophalangeal joints, the hand cutaneous network was always anastomosed with the dorsal cutaneous network. At the level of fingers, the dorsal cutaneous network was always supplied by four branches arising from the proper digital artery. Our study supported the reliability of dorsal digitometacarpal flaps, supplied by numerous palmodorsal digital anastomoses and by a rich plexiforme network joining the hand skin supply and that of the dorsal finger skin. During the procedure, we recommend limiting the surgical dissection of the flap at the level of the middle phalanx. PMID- 18404245 TI - Different expression patterns of CK2 subunits in the brains of experimental animals and patients with transmissible spongiform encephalopathies. AB - To address the possible alteration of casein kinase 2 (CK2) in transmissible spongiform encephalopathies (TSEs), the levels and patterns of CK2 in the brain tissues of hamsters or C57BL mice inoculated intracerebrally with scrapie agents 263K or 139A were evaluated by Western blots, followed by quantitative analysis. Specific semi-quantitative RT-PCR for evaluating the mRNA transcripts of CK2 subunits was performed in parallel. Compared with normal animals, the levels of CK2alpha and CK2beta in the brains of infected hamsters and mice were significantly decreased, regardless of which scrapie agent was. However, the expression of CK2alpha' or CK2alpha'/CK2alpha'' in the animals infected with agents 263K or 139A was considerably increased. Furthermore, decreases of CK2alpha and CK2beta and increases of CK2alpha'/CK2alpha'' were observed in cerebella homogenates from one familial Creutzfeldt-Jakob disease (fCJD) case and one fatal familial insomnia (FFI) case. These results suggest that alterations of CK2 subunits in brains are illness-correlative phenomena in TSEs and indicate a potential linkage of CK2 changes with the pathogenesis of prion diseases. PMID- 18404246 TI - Protease-catalysed coupling of N-protected amino acids and peptides with 4 aminoantipyrine. AB - The enzymatic synthesis of N-protected L-aminoacyl- and L-peptidyl-antipyrine amides was accomplished by proteases from different classes. Serine and cysteine proteases proved to be suitable tools for the production of amino acids and peptides conjugated to 4-aminoantipyrine, whereas metalloproteases do not seem to be very qualified for accepting this nucleophile. The product yields were optimised by applying ample opportunities of medium engineering, e.g. aqueous organic, biphasic, suspension and solid-to-solid reaction systems. Thus, yields up to 100% could be obtained. The products were purified and characterised by polarimetry and NMR spectroscopy. These results broaden the common knowledge of the catalytic potential of proteases, in particular with regard to the suitability of a special heterocyclic 1,2-amino ketone as a nucleophile for the biocatalytic amidation of amino acids and peptides. PMID- 18404248 TI - Mechanical properties of nanotubes of polyelectrolyte multilayers. AB - The elastic properties of nanotubes fabricated by layer-by-layer (LbL) assembly of polyelectrolytes in the nanopores of polycarbonate track-etched membranes have been investigated by resonant contact Atomic Force Microscopy (AFM), for nanotube diameters in the range of 100 to 200 nm. The elastic modulus of the nanotubes was computed from the resonance frequencies of a cantilever resting on freely suspended LbL nanotubes. An average value of 115 MPa was found in air for Young's modulus of these nanostructures, well below the values reported for dry, flat multilayers, but in the range of values reported for water-swollen flat multilayers. These low values are most probably due to the lower degree of ionic cross-linking of LbL nanotubes and their consequently higher water content in air, resulting from the peculiar mode of growth of nanoconfined polyelectrolyte multilayers. PMID- 18404249 TI - Simulations of two-dimensional foam rheology: localization in linear Couette flow and the interaction of settling discs. AB - Surface Evolver simulations of flowing two-dimensional foams are described. These are used for two purposes. Firstly, to extract the location of the T 1s, the changes in bubble topology that occur during plastic flow. It is shown that in linear Couette flow the T1 s are localized in space, becoming more so as the polydispersity of the foam decreases. Secondly, the sedimentation of two circular discs through a foam under gravity is studied. If the discs are sufficiently close, they begin to interact and one moves behind the other during their descent. PMID- 18404247 TI - Pathogenetic significance of aberrant glycosylation of IgA1 in IgA nephropathy. AB - IgA nephropathy (IgAN), the most common form of primary glomerulonephritis worldwide, is defined by predominant IgA1 deposits in the glomerular mesangium. Among abnormalities of the IgA immune system reported so far in IgAN, aberrant O linked glycosylation in the hinge region of IgA1 is the most consistent finding. IgA1 molecules bearing abnormal glycosylation have been found in serum, in tonsillar lymphocytes, and in eluate from mesangial deposits, and characterized by decreased O-linked N-acetylgalactosamine residues with or without alteration in the terminal sialylation of the O-linked sugars. IgA1 with incomplete galactosylation has a tendency to accumulate in glomerular mesangium by self aggregation or immune complex formation. Glomerular mesangial cells exposed to immune complexes of these IgA1 can proliferate and secrete cytokines, chemokines, growth factors, and extracellular matrix components promoting inflammatory reactions in the glomeruli. Although genes encoding enzymes involved in the O glycosylation process, such as C1GALT1, have been reported to be responsible for susceptibility to IgAN, recent evidence suggests that the abnormality is restricted to a small fraction of B cell populations and arises from dysregulated IgA1 production and secretion in mucosal immune system. This review will focus on and discuss the role of incompleteness of IgA1 O-galactosylation in the pathogenesis of IgAN and propose a possible mechanism in which abnormal IgA1 occurs in IgAN. PMID- 18404250 TI - An acidic environment changes cyclin D1 localization and alters colony forming ability in gliomas. AB - The human glioma cell lines, U87 and T98G, were evaluated for their ability to survive and form colonies in an acidic environment of pH(ext) 6.0. In contrast to U87, which showed an 80-90% survival rate, only 40% of T98G cells survived 6 days at pH(ext) 6.0 and lost their colony forming ability when returned to a normocidic environment. Although both U87 and T98G cells maintain an intracellular pH (pH(i)) of 7.0 at pH(ext) 6.0 and arrest mostly in G1 phase of the cell cycle, only T98G demonstrated a major loss of cyclin D1 that was prevented by the proteasome inhibitor MG132. Colony forming ability was restored by stably transfecting T98G cells with a cyclin D1-expressing plasmid. Both U87 and T98G cells demonstrated increased cytoplasmic localization of cyclin D1 during exposure at pH(ext) 6.0. Upon prolonged (24 h) incubation at pH(ext) 6.0, nuclear cyclin D1 was nearly absent in T98G in contrast to U87 cells. Thus, an acidic environment triggers cytoplasmic localization and proteasomal degradation of cyclin D1. PMID- 18404251 TI - Dissolution media simulating conditions in the proximal human gastrointestinal tract: an update. AB - PURPOSE: The aim of this study was to update the compositions of biorelevant media to represent the composition and physical chemical characteristics of the gastrointestinal fluids as closely as possible while providing physical stability during dissolution runs and short-term storage. METHODS: Media were designed to reflect postprandial conditions in the stomach and proximal small intestine in the "early", "middle", and "late" phases of digestion. From these "snapshot" media, general media for simulating postprandial conditions were devised. Additionally, media reflecting preprandial conditions in the stomach and small intestine were revisited. RESULTS: A set of four media is presented. A recently published medium to represent the fasted stomach, FaSSGF, needed no further revision. To simulate the postprandial stomach, a new medium, FeSSGF, is presented. Media representing the upper small intestine in the fed and fasted states were fine-tuned according to physicochemical and biochemical characteristics in vivo. All four media proved to be stable under ambient storage conditions for at least 72 h as well as under usual dissolution test conditions. CONCLUSIONS: The updated dissolution media can be used to predict formulation performance and food effects in vivo. These media are more physiologically relevant and show better physical stability than their corresponding predecessors. PMID- 18404252 TI - Scaling up the spray drying process from pilot to production scale using an atomized droplet size criterion. AB - PURPOSE: The purpose of this study was to investigate the possibility of producing identical powders in pilot and production scale spray drying equipment by matching the droplet size distributions produced by two differently sized atomizers. METHODS: Particles were prepared by spray drying solutions of acetaminophen and polyvinylpyrrolidone K-30. The success of the up-scaling was evaluated by comparing the powders in terms of particle size distribution (laser diffraction), crystallinity (XPRD) and morphology (SEM). Furthermore, the influence of process parameters on other product characteristics such as stability and residual volatile content was also evaluated. RESULTS: The spray drying experiments resulted in spherical, amorphous particles with volumetric median diameters of typically 4-10 microm for pilot scale and 4-30 microm for production scale. The results showed that particles with similar morphology and crystallinity could be produced in the two applied spray dryers. However, scale up based purely on matching droplet size distributions was not feasible. CONCLUSIONS: The scale-up criterion did not account for the differences between the droplet-drying gas mixing and residence time distribution within the two spray dryers. Therefore, production scale experiments are required in order to obtain similar product characteristics as in pilot scale. PMID- 18404253 TI - Resolution enhancement by homonuclear J-decoupling: application to three dimensional solid-state magic angle spinning NMR spectroscopy. AB - We describe a simple protocol to achieve homonuclear J-decoupling in the indirect dimensions of multidimensional experiments, and to enhance spectral resolution of the backbone Calpha carbons in the 3D NCACX experiment. In the proposed protocol, the refocusing of the Calpha-CO homonuclear J-couplings is achieved by applying an off-resonance selective pi pulse to the CO spectral region in the middle of Calpha chemical shift evolution. As is commonly used in solution NMR, a compensatory echo period is used to refocus the unwanted chemical shift evolution of Calpha spins, which takes place during the off-resonance selective pulse. The experiments were carried out on the beta1 immunoglobulin binding domain of protein G (GB1). In GB1, such implementation results in significantly reduced line widths, and leads to an overall sensitivity enhancement. PMID- 18404254 TI - A comparison between lime and alkaline hydrogen peroxide pretreatments of sugarcane bagasse for ethanol production. AB - Pretreatment procedures of sugarcane bagasse with lime (calcium hydroxide) or alkaline hydrogen peroxide were evaluated and compared. Analyses were performed using 2(3) factorial designs, with pretreatment time, temperature, and lime loading and hydrogen peroxide concentration as factors. The responses evaluated were the yield of total reducing sugars (TRS) and glucose released from pretreated bagasse after enzymatic hydrolysis. Experiments were performed using the bagasse, as it comes from an alcohol/sugar factory and bagasse, in the size, range from 0.248 to 1.397 mm (12-60 mesh). The results show that, when hexoses and pentoses are of interest, lime should be the pretreatment agent chosen, as high TRS yields are obtained for non-screened bagasse using 0.40 g lime/g dry biomass at 70 degrees C for 36 h. When the product of interest is glucose, the best results were obtained with lime pretreatment of screened bagasse. However, the results for alkaline peroxide and lime pretreatments of non-screened bagasse are not very different. PMID- 18404255 TI - Botulinum toxins revisited. PMID- 18404256 TI - Tenebrio beetles use magnetic inclination compass. AB - Animals that guide directions of their locomotion or their migration routes by the lines of the geomagnetic field use either polarity or inclination compasses to determine the field polarity (the north or south direction). Distinguishing the two compass types is a guideline for estimation of the molecular principle of reception and has been achieved for a number of animal groups, with the exception of insects. A standard diagnostic method to distinguish a compass type is based on reversing the vertical component of the geomagnetic field, which leads to the opposite reactions of animals with two different compass types. In the present study, adults of the mealworm beetle Tenebrio molitor were tested by means of a two-step laboratory test of magnetoreception. Beetles that were initially trained to memorize the magnetic position of the light source preferred, during the subsequent test, this same direction, pursuant geomagnetic cues only. In the following step, the vertical component was reversed between the training and the test. The beetles significantly turned their preferred direction by 180 degrees. Our results brought until then unknown original findings that insects, represented here by the T. molitor species, use-in contrast to another previously researched Arthropod, spiny lobster-the inclination compass. PMID- 18404257 TI - Distribution of S haplotypes and its relationship with restorer-maintainers of self-incompatibility in cultivated Brassica napus. AB - Brassica napus (AACC, 2n = 38) is a self-compatible amphidiploid plant that arose from the interspecies hybridization of two self-incompatible species, B. rapa (AA, 2n = 20) and B. oleracea (CC, 2n = 18). Self-incompatibility (S) haplotypes in one self-incompatible line and 124 cultivated B. napus lines were detected using S-locus-specific primers, and their relationships with restorer-maintainers were investigated. Two class I (S-I ( SLG ) a and S-I ( SLG ) b) and four class II (S-II ( SLG ) a, S-II ( SLG ) b, S-II ( SP11 ) a and S-II ( SP11 ) b) S haplotypes were observed, of which S-II ( SP11 ) b was newly identified. The nucleotide sequence of SP11 showed little similarity to the reported SP11 alleles. The lines were found to express a total of eleven S genotypes. The self incompatible line had a specific genotype consisting of S-II ( SP11 ) a, similar to B. rapa S-60, and S-II ( SLG ) a, similar to B. oleracea S-15. Restorers expressed six genotypes: the most common genotype contained S-I ( SLG ) a, similar to B. rapa S-47, and S-II ( SLG ) b, similar to B. oleracea S-15. Maintainers expressed nine genotypes: the predominant genotype was homozygous for two S haplotypes, S-II ( SLG ) a and S-II ( SP11 ) b. One genotype was specific to restorers and four genotypes were specific to maintainers, whereas five genotypes were expressed in both restorers and maintainers. This suggests that there is no definitive correlation between the distribution of S genotypes and restorer-maintainers of self-incompatibility. The finding that restorers and maintainers express unique genotypes, and share some common genotypes, would be valuable for detecting the interaction of S haplotypes in inter- or intra-genomes as well as for developing markers-assisted selection in self-incompatibility hybrid breeding. PMID- 18404259 TI - Out-of-hospital interventions by the French Emergency Medical Service are associated with a high survival in patients aged 80 year or over. PMID- 18404258 TI - Intravitreous anti-VEGF for diabetic retinopathy: hopes and fears for a new therapeutic strategy. AB - Vascular endothelial growth factor (VEGF) plays a key role in the development of both proliferative diabetic retinopathy (PDR) and diabetic macular oedema (DMO). In recent years, anti-VEGF agents have emerged as new approaches to the treatment of these devastating diabetic complications. Although Phase III studies in the diabetic population are needed, intravitreal anti-VEGF therapy is currently being used in clinical practice. Intravitreal injection is an effective means of delivering anti-VEGF drugs to the retina. However, this is an invasive procedure associated with potentially serious complications, such as endophthalmitis or retinal detachment, which may be significant for patients requiring serial treatment over many years. In addition, although delivered within the vitreous, anti-VEGF drugs could pass into the systemic circulation, which could potentially result in hypertension, proteinuria, increased cardiovascular events and impaired wound healing. Pegaptanib, ranibizumab and bevacizumab are the currently available anti-VEGF agents. Ranibizumab and bevacizumab block all VEGF isoforms, thus impairing both physiological and pathological neovascularisation. Pegaptanib only blocks the VEGF(165) isoform, and would therefore seem the best option for avoiding systemic adverse effects in diabetic patients, although this remains to be demonstrated in clinical trials. In this regard, head-to-head studies designed to evaluate not only the efficacy, but also the systemic adverse effects of these drugs in a high-risk population such as diabetic patients are warranted. PMID- 18404260 TI - Differential involvement of opioidergic and serotonergic systems in the antinociceptive activity of N-arachidonoyl-phenolamine (AM404) in the rat: comparison with paracetamol. AB - It is recognized that paracetamol undergoes a metabolic transformation to N arachydonylphenolamine (AM404), a CB(1) receptor ligand and anandamide uptake inhibitor. Using hot-plate and paw pressure tests, we decided to establish whether AM404 may act through opioidergic and serotonergic mechanisms. Thus, we pretreated rats with opioid mu(1) (naloxonazine) and kappa (MR2266) or 5-HT(1A) (NAN-190), 5-HT(2) (ketanserin), and 5-HT(3) (ondansetron) receptor antagonists. We investigated the possible changes in 5-hydroxyindoleacetic acid/serotonin ratio in the frontal cortex and pons. The antinociceptive effect of AM404 (10 mg/kg, intraperitoneally) or paracetamol (400 mg/kg, intraperitoneally) in either test was abolished by naloxonazine or MR2266. Ondansetron prevented AM404 activity; NAN-190 and ketanserin were ineffective. Ketanserin antagonized paracetamol activity; NAN-190 and ondansetron were inactive. AM404 did not change serotonergic activity, while paracetamol decreased serotonin turnover. The diverse antinociceptive potency of the compounds might be explained by the different influence on the serotonergic system, despite a similar involvement of opioidergic one. PMID- 18404261 TI - Development of analytical procedures for the determination of hexavalent chromium in corrosion prevention coatings used in the automotive industry. AB - The European directive 2000/53/EC limits the use of Cr(VI) in vehicle manufacturing. Although a maximum of 2 g of Cr(VI) was authorised per vehicle for corrosion prevention coatings of key components, since July 2007 its use has been prohibited except for some particular applications. Therefore, the objective of this work was to develop direct analytical procedures for Cr(VI) determination in the different steel coatings used for screws. Instead of working directly with screws, the optimisation of the procedures was carried out with metallic plates homogeneously coated to improve the data comparability. Extraction of Cr(VI) from the metallic parts was performed by sonication. Two extraction solutions were tested: a direct water extraction solution used in standard protocols and an ammonium/ammonia buffer solution at pH 8.9. The extracts were further analysed for Cr speciation by high-performance liquid chromatography (HPLC) inductively coupled plasma (ICP) atomic emission spectrometry or HPLC ICP mass spectrometry depending on the concentration level. When possible, the coatings were also directly analysed by solid speciation techniques (X-ray photoelectron spectroscopy, XPS, and X-ray absorption near-edge structure, XANES) for validation of the results. Very good results between the different analytical approaches were obtained for the sample of coating made up of a heated paint containing Zn, Al and Cr when using the extracting buffer solution at pH 8.9. After a repeated four-step extraction procedure on the same portion test, taking into account the depth of the surface layer reached, good agreement with XPS and XANES results was obtained. In contrast, for the coatings composed of an alkaline Zn layer where Cr(VI) and Cr(III) are deposited, only the extraction procedure using water allowed the detection of Cr(VI). To elucidate the Cr(VI) reduction during extraction at pH 8.9, the reactivity of Cr(VI) towards different species of Zn generally present in the coatings (metallic Zn and zinc oxide) was studied. The results showed that metallic Zn rapidly reduces Cr(VI), whereas this reaction is less evident in the presence of zinc oxide. Water was then retained for coatings containing metallic Zn. PMID- 18404262 TI - Waiting to retrieve: possible implications for brain function. AB - There is evidence that the mid-ventrolateral prefrontal cortex is involved in control processing necessary for the active retrieval of mnemonic information when retrieval cannot be triggered automatically by unique/constant associations within memory. We hypothesized that this process is initiated as soon as an instructional cue to retrieve a particular aspect of an encoded stimulus is presented prior to the appearance of the test stimulus that prompts the subjects' response. To test this hypothesis, we provided the subjects with a delay period following the presentation of the instructional cue. Across delays, the subjects took longer to respond during the active retrieval compared with the automatic retrieval trials. Importantly, for both trial types, delays of up to 300 ms improved the subjects' reaction times during the test period by an average of 101 ms. Retrieval mechanisms, therefore, are initiated early during the post instruction delay period prior to the presentation of the test stimulus needed for the decision. PMID- 18404263 TI - Image-guided drainage of multiple intraabdominal abscesses in children with perforated appendicitis: an alternative to laparotomy. AB - BACKGROUND: Appendicitis is the most common cause of an acute abdomen in children. With perforation, multiple intraperitoneal collections can be seen at presentation. In this situation, surgical treatment alone is rarely effective. OBJECTIVE: To determine the role of image-guided drainage in treating patients with acute appendicitis complicated by multiple intraabdominal collections. MATERIALS AND METHODS: A retrospective review of patient charts and interventional radiology records was performed to identify all patients with acute complicated appendicitis treated by multiple image-guided drainage procedures. Data reviewed included the number of drainages and aspirations performed, drain dwell time, the clinical course and temperature profile, and the length of inpatient hospital stay and any complications experienced. RESULTS: The study population comprised 42 children with a mean age of 107.6 months. A total of 100 drainage catheters were inserted and 56 aspirations were performed. Of the 42 children, 24 were successfully treated at a single sitting, while 18 returned for further intervention. The mean drain dwell time was 8.18 days. The mean inpatient stay was 15.02 days. Treatment of the acute presentation with image guided intervention was successful in 92.3% of children. CONCLUSION: Successful management of acute perforated appendicitis with multiple intraabdominal abscesses can be achieved with multiple minimally invasive image-guided drainage procedures. PMID- 18404264 TI - Push-me-pull-you: how microtubules organize the cell interior. AB - Dynamic organization of the cell interior, which is crucial for cell function, largely depends on the microtubule cytoskeleton. Microtubules move and position organelles by pushing, pulling, or sliding. Pushing forces can be generated by microtubule polymerization, whereas pulling typically involves microtubule depolymerization or molecular motors, or both. Sliding between a microtubule and another microtubule, an organelle, or the cell cortex is also powered by molecular motors. Although numerous examples of microtubule-based pushing and pulling in living cells have been observed, it is not clear why different cell types and processes employ different mechanisms. This review introduces a classification of microtubule-based positioning strategies and discusses the efficacy of pushing and pulling. The positioning mechanisms based on microtubule pushing are efficient for movements over small distances, and for centering of organelles in symmetric geometries. Mechanisms based on pulling, on the other hand, are typically more elaborate, but are necessary when the distances to be covered by the organelles are large, and when the geometry is asymmetric and complex. Thus, taking into account cell geometry and the length scale of the movements helps to identify general principles of the intracellular layout based on microtubule forces. PMID- 18404265 TI - Identification of functionally important amino acids in a novel indigo-producing oxygenase from Rhodococcus sp. strain T104. AB - A novel indigo-producing oxygenase gene, designated ipoA (1,197 bp) was characterized from Rhodococcus sp. strain T104. Three indigo-negative mutations (A58V, P59L, and G251D) were obtained through random mutagenesis using an E. coli mutator strain. Subsequent saturation mutagenesis resulted in the identification of nine and three amino acid substitutions that restore activity in the A58V and P59L mutants, respectively. Activity was not restored in the G251D mutation by any other amino acids. Interestingly, activity in the A58V mutant, where a methyl group is only replaced by an isopropyl side chain, is restored by a variety of amino acids, including polar ones. A molecular modeling study suggests that the residues at positions 58, 59, and 251 of the T104 IpoA enzyme are far from the active site, indicating that the mutations must alter the overall structure of the enzyme. PMID- 18404266 TI - Influence of synthetic packing materials on the gas dispersion and biodegradation kinetics in fungal air biofilters. AB - The biodegradation of toluene was studied in two lab-scale air biofilters operated in parallel, packed respectively with perlite granules (PEG) and polyurethane foam cubes (PUC) and inoculated with the same toluene-degrading fungus. Differences on the material pore size, from micrometres in PEG to millimetres in PUC, were responsible for distinct biomass growth patterns. A compact biofilm was formed around PEG, being the interstitial spaces progressively filled with biomass. Microbial growth concentrated at the core of PUC and the excess of biomass was washed-off, remaining the gas pressure drop comparatively low. Air dispersion in the bed was characterised by tracer studies and modelled as a series of completely stirred tanks (CSTR). The obtained number of CSTR (n) in the PEG packing increased from 33 to 86 along with the applied gas flow (equivalent to empty bed retention times from 48 to 12 s) and with operation time (up to 6 months). In the PUC bed, n varied between 9 and 13, indicating that a stronger and steadier gas dispersion was achieved. Michaelis-Menten half saturation constant (km) estimates ranged 71-113 mg m(-3), depending on the experimental conditions, but such differences were not significant at a 95% confidence interval. The maximum volumetric elimination rate (rm) varied from 23 to 50 g m(-3) h(-1). Comparison between volumetric and biomass specific biodegradation activities indicated that toluene mass transfer was slower with PEG than with PUC as a consequence of a smaller biofilm surface and to the presence of larger zones of stagnant air. PMID- 18404267 TI - Fast T2 mapping of the patellar articular cartilage with gradient and spin-echo magnetic resonance imaging at 1.5 T: validation and initial clinical experience in patients with osteoarthritis. AB - OBJECTIVE: To evaluate the T2 mapping of patellar articular cartilage in patients with osteoarthritis using gradient and spin-echo (GRASE) magnetic resonance (MR) imaging. MATERIALS AND METHODS: After the imaging of a phantom consisting of two sealed 50-ml test objects with different concentrations (30% and 90% weight/volume) of copper sulphate, the T2 mapping of patellar articular cartilage was performed in 35 patients (21 male and 14 female; mean age +/- SD 42+/-17 years) with moderate degree of patellar osteoarthritis. Turbo-spin-echo (TSE) (TR milliseconds/ minimum-maximum TE milliseconds 3,000/15-120; total acquisition time 5 min 52 s) and GRASE (TR milliseconds/ minimum-maximum TE milliseconds 3,000/15-120; total acquisition time 1 min 51 s) were employed. In each patient patellar cartilage was segmented at nine locations (three superior, three central, and three inferior) by manually defined regions of interest. T2 relaxation times were calculated using a linear fit applied to the logarithm of signal intensity decay. RESULTS: In the phantom the T2 values measured by GRASE were similar to those measured by MR spectroscopy (test object 1: 48.1 ms vs 51 ms; test object 2: 66.8 ms vs 71 ms; P>0.05, Wilcoxon test). In patients GRASE and TSE-derived T2 values demonstrated good agreement (mean difference +/- SD, 1.81+/-3.63 ms). The within-patient coefficient of variation was 22% for TSE and 23% for GRASE. CONCLUSION: Fast T2 mapping of the patellar articular cartilage can be performed with GRASE within a third of the time of that of standard sequences. PMID- 18404268 TI - The potential of 211Astatine for NIS-mediated radionuclide therapy in prostate cancer. AB - PURPOSE: We reported recently the induction of selective iodide uptake in prostate cancer cells (LNCaP) by prostate-specific antigen (PSA) promoter directed sodium iodide symporter (NIS) expression that allowed a significant therapeutic effect of (131)I. In the current study, we studied the potential of the high-energy alpha-emitter (211)At, also transported by NIS, as an alternative radionuclide after NIS gene transfer in tumors with limited therapeutic efficacy of (131)I due to rapid iodide efflux. METHODS: We investigated uptake and therapeutic efficacy of (211)At in LNCaP cells stably expressing NIS under the control of the PSA promoter (NP-1) in vitro and in vivo. RESULTS: NP-1 cells concentrated (211)At in a perchlorate-sensitive manner, which allowed a dramatic therapeutic effect in vitro. After intraperitoneal injection of (211)At (1 MBq), NP-1 tumors accumulated approximately 16% ID/g (211)At (effective half-life 4.6 h), which resulted in a tumor-absorbed dose of 1,580+/-345 mGy/MBq and a significant tumor volume reduction of up to 82+/-19%, while control tumors continued their growth exponentially. CONCLUSIONS: A significant therapeutic effect of (211)At has been demonstrated in prostate cancer after PSA promoter directed NIS gene transfer in vitro and in vivo suggesting a potential role for (211)At as an attractive alternative radioisotope for NIS-targeted radionuclide therapy, in particular in smaller tumors with limited radionuclide retention time. PMID- 18404269 TI - Carotid artery wall thickness and ischemic symptoms: evaluation using multi detector-row CT angiography. AB - The purpose of this study was to determine if carotid artery wall thickness (CAWT) studied by using multi-detector-row CT angiography (MDCTA) can be considered an effective parameter predictive of increased risk of stroke. A total of 217 patients were retrospectively studied by using MDCTA. In all patients CAWT was measured with an internal digital caliper. Continuous data were described as the mean value +/- standard deviation (SD), and they were compared with Student's t-test. Scatter plots to determine interobserver agreement were performed, and correlation coefficient was calculated with Pearson statistics. A P value <0.05 was considered to mean statistical significance. Measurements of the distal common CAWT ranged from 0.5 to 1.6 mm. In the patient group without stroke, average CAWT was 0.82 mm (0.22 SD), whereas in patient group with stroke it was 1.096 mm (0.21 SD). CAWT in patients without stroke showed statistical difference (P < 0.0001) when compared to patients with stroke. By using a threshold of 1 mm, an important statistical association between thick CAWT and stroke was found (P < 0.0001). In fact, patients with > or = 1 mm CAWT had stroke with an odds ratio of 8.16 when compared with patients with <1 mm CAWT. Resulting data suggested that an increased CAWT is an indicator for risk of stroke. This parameter should be considered in addition to other well-known risk factors such as diabetes, hypertension, smoking, and dyslipidemia. PMID- 18404271 TI - Protective effects of phenolic fraction of blue honeysuckle fruits against UVA induced damage to human keratinocytes. AB - In this study, the UVA (photo)protective activity of the phenolic fraction of L. caerulea fruits (PFLC) was assessed in human keratinocytes HaCaT. The keratinocytes were pre- or post-treated with PFLC (1-250 mg/l) and exposed to UVA irradiation (10-30 J/cm(2)). The results showed that both pre- and post-treatment with PFLC significantly suppressed UVA-induced ROS production, which was also revealed as a decrease in intracellular lipid peroxidation and elevation of reduced glutathione. Protection was concentration-dependent with a maximum at 50 mg/l. These results suggest that PFLC attenuates UVA-induced oxidative stress by reduction of ROS generation and ROS-mediated damage. For this reason, PFLC has potentially skin-protective functions against the deleterious effects of sunlight. PMID- 18404270 TI - Invertebrate trypsins: a review. AB - Food protein hydrolysis, a crucial step in digestion, is catalyzed by trypsin enzymes from the digestive apparatus of invertebrates. Trypsin appeared early in evolution and occurs in all phyla and, in the digestive systems of invertebrates, it became the most abundant proteinase. As in vertebrates, invertebrate trypsin is also present in several forms (isoenzymes). Its physiological importance in food protein digestion in several invertebrate species has emerged with compelling evidence; and several other physiological functions, such as regulation of digestive functions, are now settled. Recent advances in the knowledge of invertebrate trypsin synthesis, regulation, genetics, catalytic characteristics; structure, evolution, as well as inhibition, especially in non Drosophilidae insects and in some crustaceans are reviewed. Most of the existing information is largely based on the use of several tools, including molecular techniques, to answer many still open questions and solve medical, agricultural, and food quality problems. PMID- 18404272 TI - Ultraviolet-B and water stress effects on growth, gas exchange and oxidative stress in sunflower plants. AB - The effects and interaction of drought and UV-B radiation were studied in sunflower plants (Helianthus annuus L. var. Catissol-01), growing in a greenhouse under natural photoperiod conditions. The plants received approximately 1.7 W m( 2) (controls) or 8.6 W m(-2) (+UV-B) of UV-B radiation for 7 h per day. The UV-B and water stress treatments started 18 days after sowing. After a period of 12 days of stress, half of the water-stressed plants (including both UV-B irradiated or non-irradiated) were rehydrated. Both drought and UV-B radiation treatments resulted in lower shoot dry matter per plant, but there was no significant interaction between the two treatments. Water stress and UV-B radiation reduced photosynthesis, stomatal conductance and transpiration. However, the amplitude of the effects of both stressors was dependent on the interactions. This resulted in alleviation of the negative effect of drought on photosynthesis and transpiration by UV-B radiation as the water stress intensified. Intercelluar CO(2) concentration was initially reduced in all treatments compared to control plants but it increased with time. Photosynthetic pigments were not affected by UV-B radiation. Water stress reduced photosynthetic pigments only under high UV-B radiation. The decrease was more accentuated for chlorophyll a than for chlorophyll b. As a measure for the maximum efficiency of photosystem II in darkness F (v)/F (m) was used, which was not affected by drought stress but initially reduced by UV-B radiation. Independent of water supply, UV-B radiation increased the activity of pirogalol peroxidase and did not increase the level of malondialdehyde. On the other hand, water stress did not alter the activity of pirogalol peroxidase and caused membrane damage as assessed by lipid peroxidation. The application of UV-B radiation together with drought seemed to have a protective effect by lowering the intensity of lipid peroxidation caused by water stress. The content of proline was not affected by UV-B radiation but was increased by water stress under both low and high UV-B radiation. After 24 h of rehydration, most of the parameters analyzed recovered to the same level as the unstressed plants. PMID- 18404273 TI - Non-arteritic anterior ischemic optic neuropathy: role of systemic corticosteroid therapy. AB - OBJECTIVE: To investigate systematically the role of systemic corticosteroid therapy in non-arteritic anterior ischemic optic neuropathy (NA-AION). METHODS: The study consists of a cohort of 613 consecutive patients (696 eyes), first seen in our clinic from 1973 to 2000. Of this cohort, 312 patients (364 eyes) voluntarily opted for systemic steroid therapy, and 301 (332 eyes) for no treatment. At first visit, all patients in both groups had a detailed ophthalmic and medical history, and comprehensive ophthalmic evaluation. Visual evaluation was done by recording Snellen visual acuity, and visual fields with a Goldmann perimeter. The same ophthalmic evaluation was performed at each follow-up visit. Patients in the steroid-treated group were initially given 80 mg Prednisone daily for 2 weeks, and then tapered down to 70 mg for 5 days, 60 mg for 5 days, and then cutting down by 5 mg every 5 days. Visual outcome in the two groups was compared RESULTS: Median follow-up was 3.8 years. At 6 months from onset of NA AION, of the eyes with initial visual acuity 20/70 or worse and seen within 2 weeks of onset, there was visual acuity improvement in 69.8% (95% confidence interval (CI): 57.3%, 79.9%) in the treated group, compared to 40.5% (95% CI: 29.2%, 52.9%) in the untreated group (odds ratio of improvement: 3.39; 95% CI:1.62, 7.11; p = 0.001). Comparison of visual field defect at 6 months from onset of NA-AION, among those seen within 2 weeks of NA-AION onset with moderate to severe initial visual field defect, there was improvement in 40.1% (95% CI: 33.1%, 47.5%) of the treated group, and 24.5% (95% CI: 17.7%, 32.9%) of the untreated group (odds ratio: 2.06, 95% CI: 1.24, 3.40; p = 0.005). In both treated and untreated groups, the visual acuity and visual fields kept improving up to about 6 months from onset of NA-AION, and very little thereafter. CONCLUSION: This study suggested that NA-AION eyes treated during the acute phase with systemic corticosteroids resulted in a significantly higher probability of improvement in visual acuity (p = 0.001) and visual field (p = 0.005) than in the untreated group. Both visual acuity and visual fields improved up to 6 months after onset of NA-AION. PMID- 18404274 TI - Case of retinochoroiditis associated with porcine ascarids diagnosed by antibodies in subretinal fluid. AB - PURPOSE: To determine the cause of retinochorioditis in a patient with a granulomatous retinal exudate and an exudative retinal detachment. CASE: A 45 year-old man presented at another hospital with increased visual disturbances of the left eye. He was diagnosed with uveitis, and treated with topical steroids for 1 month. However, the uveitis worsened, and he was referred to our hospital. Ophthalmoscopy showed a yellowish-white granulomatous exudate, and an exudative retinal detachment in the lower peripheral retina. The retinal detachment worsened and affected the macula. Pars plana vitrectomy was performed, and the retina was reattached. During the surgery, ocular samples were collected for further examinations. The titers of antibodies against 12 kinds of ascaridis were examined, and elevated titers of specific antibodies against porcine ascarids were detected in the subretinal fluid, but not in the aqueous humor, vitreous, or serum. CONCLUSIONS: Vitrectomy with the collection of ocular samples, especially subretinal fluid, was a key procedure in the diagnosis and treatment of retinochoroiditis associated with the porcine ascarids. PMID- 18404275 TI - Reproductive outcome in a cohort of male and female rubber workers: a registry study. AB - PURPOSE: To investigate whether blue-collar employment in the Swedish rubber industry from 1973 onwards had a negative impact on reproductive health. METHODS: Pairs of mother and child, and triads of father-mother-child were obtained through linkage of a cohort of 18,518 rubber factory employees with the Swedish Population Registry. Birth outcomes were obtained from the Medical Birth Register for 17,918 children. For each child, parental employment as blue-collar rubber worker during the pregnancy and sperm maturation period was obtained from work place records. Children to female food industry workers, in all 33,256, constituted an external reference group. RESULTS: The sex ratio was reversed, with odds ratio (OR) for having a girl was 1.15 (95% CI 1.02, 1.31) when the mother was exposed. When both parents were exposed, the OR was even higher, 1.28 (95% CI 1.02, 1.62). An increased risk of multiple births was observed when both parents were exposed, with OR 2.42 (95% CI 1.17, 5.01). Children with both maternal and paternal exposure had a reduced birth weight compared to the external reference cohort. After adjustment for smoking (available for births from 1983 onwards), ethnicity and sex, the difference between children (singletons, live births) with maternal and paternal exposure and external referents was -142 g (95% CI -229, -54). The adjusted OR for having a small-for gestational-age child was 2.15 (95% CI 1.45, 3.18) when the mother was a rubber worker during the pregnancy. CONCLUSION: There were clear indications that reproductive outcome was adversely affected in rubber workers. The findings warrant further investigation with refinement of exposure indices and inclusion of other endpoints of reproductive health. PMID- 18404276 TI - Noise induced hearing loss and other hearing complaints among musicians of symphony orchestras. AB - OBJECTIVES: An investigation of the hearing status of musicians of professional symphony orchestras. Main questions are: (1) Should musicians be treated as a special group with regard to hearing, noise, and noise related hearing problems (2) Do patterns of hearing damage differ for different instrument types (3) Do OAE have an added value in the diagnosis of noise induced hearing loss (NIHL) in musicians. METHODS: 241 professional musicians, aged between 23-64 participated. A brief medical history and the subjective judgment of their hearing and hearing problems were assessed. Musicians were subjected to an extensive audiological test battery, which contained testing of audiometric thresholds, loudness perception, diplacusis, tinnitus, speech perception in noise, and otoacoustic emissions. RESULTS: Most musicians could be categorized as normal hearing, but their audiograms show notches at 6 kHz, a frequency that is associated with NIHL. Musicians often complained about tinnitus and hyperacusis, while diplacusis was generally not reported as a problem. Tinnitus was most often localized utmost left and this could not be related to the instrument. It was usually perceived in high frequency areas, associated with NIHL. In general, musicians scored very well on the speech-in-noise test. The results of the loudness perception test were within normal limits. Otoacoustic emissions were more intense with better pure-tone thresholds, but due to large individual differences it can still not be used as an objective test for early detection of NIHL. CONCLUSIONS: Musicians show more noise induced hearing loss than could be expected on the basis of age and gender. Other indicators, such as complaints and prevalence of tinnitus, complaints about hyperacusis and prevalence of diplacusis suggest that musicians' ears are at risk. Continuing education about the risks of intensive sound exposure to musicians, with the emphasis on the possible development of tinnitus and hyperacusis and the need for good hearing protection is warranted. PMID- 18404277 TI - Characteristics of well-differentiated thyroid cancer associated with multinodular goiter. AB - BACKGROUND: The likelihood of thyroid cancer is similar in patients with one or more nodules, but tumor characteristics of thyroid cancer in glands with multinodular goiter are unclear. MATERIALS AND METHODS: This study consisted of 100 consecutive patients (ages 15-81 years; 84 women), who underwent thyroidectomy for papillary or follicular thyroid carcinoma between 2005 and 2006. Patient demographics, diagnostic tests, operations, and pathological findings were reviewed. RESULTS: In a univariate analysis, cancer diagnosed within multinodular goiter was different from solitary cancer nodule in age of diagnosis (48 vs 40 years, p = 0.002), tumor size (1.42 vs 2.20 cm, p = 0.024), and the presence of cervical lymph node metastases (p = 0.035). There was no difference in gender, extrathyroidal invasion, multifocality, and types of operation. Only age (odds ratio, 1.056) and tumor size (odds ratio, 0.730) revealed independent correlation in multivariate analysis. CONCLUSION: Thyroid cancer in glands with multiple nodules is associated with older age at diagnosis and smaller tumor size. PMID- 18404278 TI - The effect of combination therapy with albendazole and praziquantel on hydatid cyst treatment. AB - The standard therapy of hydatid cyst is surgery but, in nonoperable patients and multiple organ involvement, medical therapy may be more useful. The efficacy of drugs especially in short-term treatment of hydatid cyst is unknown. This study was carried out to evaluate the effect of combination therapy with albendazole and praziquantel in the treatment of hydatid diseases. In a nonrandomized quasi experimental study, nine patients with multiple hydatid cysts were treated with albendazole (400 mg/twice a day) and praziquantel (40 mg/kg per day) twice a week for 4 weeks. This regimen was repeated for three courses with a 2-week interval between each one. The average follow-up period after treatment was 18 months. Response to treatment was assessed through the observation of the symptoms and radiologic findings (computed axial tomography scan, sonography, X-ray). Symptoms disappeared in seven (77.8%) patients and improved partially in two (22.2%) patients. Radiological assessment showed significant improvement in five (55.6%) and partial improvement in four (44.4%) patients. Combination therapy with albendazole and praziquantel is effective in the treatment of hydatid cyst and can be used as an alternative to surgery in disseminated and nonoperable cases. PMID- 18404279 TI - Refined mapping of X-linked reticulate pigmentary disorder and sequencing of candidate genes. AB - X-linked reticulate pigmentary disorder with systemic manifestations in males (PDR) is very rare. Affected males are characterized by cutaneous and visceral symptoms suggestive of abnormally regulated inflammation. A genetic linkage study of a large Canadian kindred previously mapped the PDR gene to a greater than 40 Mb interval of Xp22-p21. The aim of this study was to identify the causative gene for PDR. The Canadian pedigree was expanded and additional PDR families recruited. Genetic linkage was performed using newer microsatellite markers. Positional and functional candidate genes were screened by PCR and sequencing of coding exons in affected males. The location of the PDR gene was narrowed to a approximately 4.9 Mb interval of Xp22.11-p21.3 between markers DXS1052 and DXS1061. All annotated coding exons within this interval were sequenced in one affected male from each of the three multiplex families as well as one singleton, but no causative mutation was identified. Sequencing of other X-linked genes outside of the linked interval also failed to identify the cause of PDR but revealed a novel nonsynonymous cSNP in the GRPR gene in the Maltese population. PDR is most likely due to a mutation within the linked interval not affecting currently annotated coding exons. PMID- 18404280 TI - Emergency percutaneous treatment in iatrogenic hepatic arterial injuries. AB - The purpose of this report is to present our experience in the emergency percutaneous treatment of patients with iatrogenic hepatic arterial injury (IHAI). Seven patients (six men, one woman; age range 19-78 years; mean age 63 years) with IHAI secondary to surgical or percutaneous procedures were treated with endovascular percutaneous procedures. We performed six transhepatic arterial embolization (TAE) and one placement of an endograft stent. Follow-up was carried out by ultrasound (US) or computed tomography (CT) after 1, 3, 6, and 12 months, yearly thereafter and by laboratory tests in the first 6 months to completely exclude occult and asymptomatic bleeding. All procedures had 100% technical success. No case of further bleeding was seen at follow-up. In one patient, we detected one abscess after 3 weeks, which was treated by antibiotic therapy. In conclusion, endovascular treatment currently represents a valid option in emergency settings, as it the enables diagnosis and treatment of IHAI in a single session. PMID- 18404282 TI - Hybrid therapy for hypoplastic left heart syndrome: reality check. PMID- 18404283 TI - Hybrid therapy for hypoplastic left heart syndrome: system-wide approach is vital. PMID- 18404284 TI - Neonatal renal vein thrombosis: grey-scale and Doppler ultrasonic features. AB - Renal vein thrombosis RVT is the most common non-catheter related venous thromboembolic events VTE in newborns and is responsible for approximately 10% of all VTE in newborns. Almost 80% of all RVT present within the first month and usually within the first week of life. Currently ultrasound is the radiographic test of choice because of its practicality, sensitivity and lack of adverse effects. The sonographic features vary according to the severity, the extent of the thrombus, the development of collateral circulation and the stage of renal vein thrombosis. Initial diagnostic features include, renal enlargement, echogenic medullary streaks that have a vascular or perivascular distribution, thrombus in the vein prominent echopoor medullary pyramids, subsequently loss of corticomedullary differentiation, reduced echogenicity around the affected pyramids and echogenic band at the extreme apex of the pyramid. Higher resistance index and absent, steady, or less pulsatile venous flow on the affected side compared with flow in the contralateral kidney are helpful Doppler signs. While sonography may be useful in revealing the venous and renal morphology, it is insufficiently reliable for assessment of the functional impact of these morphologic findings. There are no characteristic grey-scale or Doppler ultrasound prognostic features to predict outcome of neonatal RVT and long-term follow up of those children is required. PMID- 18404285 TI - Pancreatic metastasis of renal cell carcinoma: multidetector CT findings. AB - Pancreatic metastasis of renal cell carcinoma is more prevalent than the rates reported in the literature before the development of multidetector computed tomography (MDCT). These lesions are usually asymptomatic and used to be an incidental finding detected on radiological follow-up of these patients. If they are not associated with extrapancreatic involvement, they show an excellent response to surgical treatment. Thus, radiologists should be aware of the radiological characteristics of this condition to allow detection at an early stage. In this article we review the MDCT features of four cases of pancreatic metastasis of renal cell carcinoma to familiarize radiologists with this entity and the most suitable study method for its detection. The main imaging findings observed were hypervascular lesions in arterial phase of biphasic studies, hence the detection of these lesions is performed better during the early phase scanning. PMID- 18404286 TI - Surgical treatment of tertiary hyperparathyroidism: the choice of procedure matters: surgical management of tertiary hyperparathyroidism: an enigma. PMID- 18404287 TI - Initial resection of potentially viable tissue is not optimal treatment for grades II-IV pancreatic injuries. AB - PURPOSE: This study was designed to ascertain the optimal therapy and diagnostics for children with pancreatic injury. METHODS: From January 1, 2001 to January 1, 2007, all children (newborn to 17 years) who presented to this Level I trauma center with demonstrated pancreatic injury were prospectively entered into the TRACS IV system and reviewed for injury type, diagnostics, therapy, demographics, and outcome. RESULTS: Fourteen children sustained grade II or higher pancreatic injury during this period. CT scan was performed for diagnosis in all cases. There were 11 boys and 3 girls, and mean age was 6.9 (range, 2-16) years. There were five grade II injuries, four grade III injuries, four grade IV injuries, and one grade V injury. All grade II injuries were treated successfully nonoperatively with observation. The nine grade III-IV injuries all underwent operative external drainage without pancreatectomy or stent placement. The single grade V injury died of multiple associated injuries after operative intervention. No pseudocysts developed in these children. All children have normal pancreatic function, and all except one have normal anatomy on follow-up scans. Early exploration and drainage directly reduces length of stay. CONCLUSION: Grade II pancreatic injuries do not require routine surgical exploration in children. Grade III and IV injuries in this series were treated with expeditious drainage of the pancreatic bed and did not require routine pancreatectomy or endoscopic stent [corrected] placement as some have recommended. Early drainage shortens hospital stay, and outcomes from this therapy are excellent. Pancreatic resection of exocrine defunctionalized segments of pancreas may be performed safely electively after acute injury if necessary, but anecdotal information from this series indicates that too may not be necessary. Grade V injuries often are accompanied by multiple other organ injuries and are associated with a significant mortality rate. A multi-institutional investigation is warranted to reassess optimal therapy for pancreatic injury in children. PMID- 18404288 TI - Risk factors for wound infection after surgery for colorectal cancer. AB - BACKGROUND: Among complications after surgery for colorectal cancer, wound infections may prolong hospitalization and increase healthcare costs. This study was designed to clarify the incidence, risk factors, and pathogens responsible for wound infections after surgery for colorectal cancer. METHODS: The study group comprised 144 patients (94 men and 50 women) with colorectal cancer in whom the same surgeon at Kitasato University Hospital performed resection from January 2004 through December 2005. Their mean age was 67.1 years (range = 38-90). To identify risk factors for surgical wound infections, we examined the following 11 variables: gender, age (>65 vs. 25 vs. 180 vs. 120 vs. O-->7',4 beta-->8') epicatechin-3'-O-gallate (8), together with (-)-epicatechin-3-O-gallate (4), (-) epigallocatechin (5), (-)-epigallocatechin-3-O-gallate (6), and (+)-gallocatechin (4 alpha-->8')-epigallocatechin (7), were isolated from the tea plant Camellia sinensis (L.) O. Kuntze var. sinensis (cv., Yabukita). The structure of 8, including stereochemistry, was elucidated by spectroscopic methods and hydrolysis. The compounds, along with commercially available pyrogallol (1), (+) catechin (2), and (-)-epicatechin (3), were examined for toxicity towards egg bearing adults of Caenorhabditis elegans. The anthelmintic mebendazole (9) was used as a positive control. Neither 2 nor 3 were toxic but the other compounds were toxic in the descending order 8, 7 approximately 6, 9, 4, 5, 1. The LC(50) (96 h) values of 8 and 9 were evaluated as 49 and 334 micromol L(-1), respectively. These data show that many green tea polyphenols may be potential anthelmintics. PMID- 18404317 TI - 5-Hydroxymethyl-2-furfural, a clinical trials agent for sickle cell anemia, and its mono/di-glucosides from classically processed steamed Rehmanniae Radix. AB - Rehmanniae Radix (Di Huang) is one of the most important traditional Chinese medicines (TCM), and is used for multiple therapeutic purposes. In our investigation of the chemical constituents of Rehmanniae Radix, steamed roots were prepared by the classical processing method. Reversed-phase HPLC of the 50% MeOH extract of steamed Rehmanniae Radix yielded three 5-hydroxymethylfurfural derivatives. The new furfural disaccharide 5-(alpha-D-glucopyranosyl-(1-->6) alpha-D-glucopyranosyloxymethyl)-2-furancarboxaldehyde (1) was isolated and characterized, together with its known aglycone 5-hydroxymethyl-2-furfural (3), which is currently in sickle cell anemia Phase I clinical trials, and its corresponding monosaccharide 5-(alpha-D-glucopyranosyloxymethyl)-2 furancarboxaldehyde (2), which was isolated as a natural product for the first time. The presence of these three compounds, particularly 3, which were not found in the unprocessed extract of Rehmanniae Radix, could substantiate the traditional medicinal use of steamed Rehmanniae Radix. PMID- 18404316 TI - Constituents from leaves of Apocynum venetum L. AB - An analysis using HPLC-MS revealed that an extract from dried leaves of Apocynum venetum L. contained more than 15 kinds of phenolic constituents. Two malonated flavonol glycosides were further isolated, and their structures were determined to be quercetin 3-O-(6''-O-malonyl)-beta-D-glucoside (1) and quercetin 3-O-(6''-O malonyl)-beta-D-galactoside (2) by NMR spectroscopic analysis. This is the first report describing the isolation of these malonated flavonol glycosides from A. venetum L. Both glycosides showed strong scavenging activity against 1, 1 diphenyl-2-picrylhydrazyl (DPPH) radical. PMID- 18404318 TI - Cesdiurins I-III, steroidal saponins from Cestrum diurnum L. AB - Phytochemical investigation of a methanolic extract of leaves of Cestrum diurnum L (Solanaceae) resulted in isolation of several furostanol steroidal saponins, named cesdiurins I-III (1-3). Their structures of the isolated compounds were determined by spectroscopic analyses, including by use of 1D and 2D NMR spectroscopic techniques as well as by mass spectral analyses. PMID- 18404319 TI - Anticarcinogenic compounds in the Uzbek medicinal plant, Helichrysum maracandicum. AB - An ethanol extract of Helichrysum maracandicum showed antiproliferative activity against cultured cells of SENCAR mouse in an in vitro assay, and activity-guided fractionation of the extract resulted in the isolation of isosalipurposide as an active substance. Naringenin chalcone, the aglycone of isosalipurposide, also showed strong antiproliferative activity. An in vivo assay of two-stage carcinogenesis on mouse skin revealed that epidermal application of isosalipurposide resulted in delayed formation of papillomas. Western blot analysis showed that the expression of p38 mitogen-activated protein kinase was suppressed by the administration of naringenin chalcone or isosalipurposide, which might be related to the anticarcinogenic activity. PMID- 18404320 TI - Antinociceptive, anti-inflammatory, and antipyretic properties of an aqueous extract of Dicranopteris linearis leaves in experimental animal models. AB - This study was performed out to establish the antinociceptive, anti-inflammatory, and antipyretic properties of an aqueous extract of Dicranopteris linearis leaves in experimental animals. The antinociceptive activity was measured using the abdominal constriction, hot plate, and formalin tests. The anti-inflammatory and antipyretic activities were measured using the carrageenan-induced paw edema and brewer's yeast-induced pyrexia tests, respectively. The extract, obtained after 72 h soaking of the air-dried leaves in distilled water and then prepared in the doses of 13.2, 66.0, 132.0, and 660.0 mg/kg, was administered subcutaneously 30 min before subjecting the animals to the assays mentioned above. Generally, the extract, at all doses used, was found to have significant (P < 0.05) concentration-independent antinociceptive, anti-inflammatory, and anti-pyretic activity. In conclusion, the aqueous extract of D. linearis has antinociceptive, anti-inflammatory, and antipyretic activity, supporting previous claims of its traditional use by the Malays to treat various ailments, particularly fever. PMID- 18404321 TI - Efficacy of extracting solvents to chemical components of kava (Piper methysticum) roots. AB - The chemical composition of kava (Piper methysticum) lactones and various phytochemicals obtained following the sonication of ground kava roots extracted in the solvents hexane, chloroform, acetone, ethanol, methanol and water, respectively, was analyzed. Eighteen kava lactones, cinnamic acid bornyl ester and 5,7-dimethoxy-flavanone, known to be present in kava roots, were identified, and seven compounds, including 2,5,8-trimethyl-1-naphthol, 5-methyl-1-phenylhexen 3-yn-5-ol, 8,11-octadecadienoic acid-methyl ester, 5,7-(OH)(2)-4'-one-6,8 dimethylflavanone, pinostrobin chalcone and 7-dimethoxyflavanone-5-hydroxy-4', were identified for the first time. Glutathione (26.3 mg/g) was found in the water extract. Dihydro-5,6-dehydrokavain (DDK) was present at a higher level than methysticin and desmethoxyyagonin, indicating that DDK is also a major constituent of kava roots. Acetone was the most effective solvent in terms of maximum yield and types of kava lactones isolated, followed by water and chloroform, whereas hexane, methanol, and ethanol were less effective as solvents. Total phenolic and antioxidant activity varied among the extracting solvents, with acetone and chloroform producing the highest effects, followed by water, while methanol, ethanol and hexane were less effective. PMID- 18404322 TI - Oligostilbenoids from Shorea gibbosa and their cytotoxic properties against P-388 cells. AB - A new oligostilbenoid derivative, diptoindonesin F (1), along with five known oligostilbenoids, (-)-ampelopsin A (2), (-)-alpha-viniferin (3), ampelopsin E (4), (-)-vaticanol B (5), and (-)-hemsleyanol D (6), were isolated from the methanol extract of the tree bark of Shorea gibbosa. The structure of the new compound was determined based on the analysis of spectroscopic data, including UV, IR, NMR 1-D and 2-D, and mass spectra. Cytotoxic properties of the isolated oligostilbenoids were evaluated against murine leukemia P-388 cells with the result that compounds 2 and 4 showed the highest cytotoxicity. PMID- 18404323 TI - A new acylphloroglucinol glycoside from Solidago altissima L. AB - A new acylphloroglucinol glycoside was isolated from the leaves of Solidago altissima L. The chemical structure of the glycoside, which has a phloroglucinol moiety with a butyryl side chain, was elucidated based on the analysis of spectroscopic data. PMID- 18404324 TI - Anti-allergic effect of a combination of Citrus unshiu unripe fruits extract and prednisolone on picryl chloride-induced contact dermatitis in mice. AB - Effect of 50% ethanolic extract of unripe fruits of Citrus unshiu (CU-ext) on type IV allergic reaction was examined by inhibitory activity of ear swelling of picryl chloride-induced contact dermatitis (PC-CD) in mice. Oral administration of CU-ext and subcutaneous administration of prednisolone showed inhibition of ear swelling during both induction and effector phases of PC-CD. The inhibitory activities of combinations of CU-ext (p.o.) and prednisolone (s.c.) during induction phase of PC-CD were more potent than those of CU-ext alone and prednisolone alone. Successive oral administration of hesperidin, a major flavanone glycoside of CU-ext, inhibited ear swelling during induction phase of PC-CD. The inhibitory activities of combinations of hesperidin (p.o.) and prednisolone (s.c.) were more potent than those of hesperidin alone and prednisolone alone. These results indicated that the combinations of prednisolone and CU-ext or hesperidin exerted a synergistic effect. PMID- 18404325 TI - Studies on the constituents from the fruits of Phaleria macrocarpa. AB - From the fruits of Phaleria macrocarpa, icariside C(3) (1), phalerin (2), and mangiferin (3) were isolated and their structures were identified on the basis of spectroscopic data. Icariside C(3) (1) showed a slow vasorelaxant activity against noradrenaline-induced contraction of isolated rat aorta. The structure of phalerin (2) was revised as 2,4',6-trihydroxy-4-methoxybenzophenone-2-O-beta-D glucoside. PMID- 18404326 TI - An efficient method of propagation of Podophyllum hexandrum: an endangered medicinal plant of the Western Himalayas under ex situ conditions. AB - This study shows an effective but simple method of conserving characterized populations and elite clones through vegetative propagation and genetic diversity through seeds in Podophyllum hexandrum (family Berberidaceae). Seed dormancy has been considered to be a major constraint in these seeds and most of the earlier reports recommended dormancy-breaking pretreatments such as chilling, gibberellic acid (GA(3)), etc. However, seeds of the 14 accessions that we tested exhibited no dormancy and hence did not require any pretreatments. Besides accession, collection of seeds with high moisture content could be one of the reasons for lack of dormancy. Thus, we propose germination of seeds (while they still retained moisture) in sand at 25 degrees C for high and reproducible results within a shorter period of time compared with earlier reports. Hypocotyl dormancy is known to considerably delay plant establishment and hence en masse propagation by preventing the emergence of functional leaves for up to 11-12 months. Manual removal of cotyledonary leaves, being labor and time intensive, is not a feasible method for large-scale seedling establishment. However, in this study, we showed that GA(3) at 200 ppm can alleviate hypocotyl dormancy besides reducing the time taken for true or functional leaf emergence. Treatment of cotyledonary leaves of 1 week-old-seedlings with 200 ppm GA(3 )resulted in true or functional leaf emergence within 7 days, and the resultant plants were also more vigorous than the ones obtained from manual removal of cotyledonary leaves. The study helped advance the establishment of seedlings by one growing season (almost 1 year). PMID- 18404327 TI - In vitro antimalarial activity of prenylated flavonoids from Erythrina fusca. AB - Ethyl acetate (EtOAc) extract from the stem bark of Erythrina fusca showed a antimalarial activity against the multi-drug-resistant strain (K1) of Plasmodium falciparum, and six flavonoids, lupinifolin (1), citflavanone (2), erythrisenegalone (3), lonchocarpol A (4), liquiritigenin (5), and 8 prenyldaidzein (6), were isolated from the extract. Diprenylated flavanone 4 showed a notable antimalarial activity (IC(50); 1.6 microg/mL); however 1 and 3 did not show the activity, even though these compounds possessed prenylated substitution. PMID- 18404328 TI - Antinociceptive and anti-inflammatory activities of the aqueous extract of Kaempferia galanga leaves in animal models. AB - This study was performed to determine the antinociceptive and anti-inflammatory activities of aqueous extract of Kaempferia galanga leaves using various animal models. The extract, in the doses of 30, 100, and 300 mg/kg, was prepared by soaking (1:10; w/v) the air-dried powdered leaves (40 g) in distilled water (dH(2)O) for 72 h and administered subcutaneously in mice/rats 30 min prior to the tests. The extract exhibited significant (P < 0.05) antinociceptive activity when assessed using the abdominal constriction, hot-plate and formalin tests, with activity observed in all tests occurring in a dose-dependent manner. Furthermore, the antinociceptive activity of K. galanga extract was significantly (P < 0.05) reversed when prechallenged with 10 mg/kg naloxone. The extract also produced a significantly (P < 0.05) dose-dependent anti-inflammatory activity when assessed using the carrageenan-induced paw-edema test. In conclusion, this study demonstrated that K. galanga leaves possessed antinociceptive and anti inflammatory activities and thus supports the Malay's traditional uses of the plant for treatments of mouth ulcer, headache, sore throat, etc. PMID- 18404329 TI - A new Erythrina alkaloid from Erythrina herbacea. AB - A new Erythrina alkaloid, 10-hydroxy-11-oxoerysotrine (1), has been isolated from the flowers of Erythrina herbacea together with five known compounds: erytharbine (2), 10,11-dioxoerysotrine (3), erythrartine (4), erysotramidine (5) and erysotrine-N-oxide (6). The structure of the new compound was elucidated on the basis of its spectral data, including 2-D NMR and mass (MS) spectra. The new compound is a rare C-10 oxygenated Erythrina alkaloid. The antioxidant activities of the isolated compounds 1-6 were evaluated by scavenging with peroxynitrite. PMID- 18404330 TI - Antitubercular triterpenes and phytosterols from Pandanus tectorius Soland. var. laevis. AB - Bioassay-guided chromatographic purification of the antitubercular chloroform extract of Pandanus tectorius Soland. var. laevis leaves afforded a new tirucallane-type triterpene, 24,24-dimethyl-5 beta-tirucall-9(11),25-dien-3-one (1), squalene and a mixture of the phytosterols stigmasterol and beta-sitosterol. Microplate Alamar Blue Assay (MABA) showed that 1 inhibited the growth of Mycobacterium tuberculosis H(37)Rv with a MIC of 64 microg/mL, while squalene and the sterol mixture have MICs of 100 and 128 microg/mL, respectively. PMID- 18404331 TI - New alkaloid from the aerial parts of Codonopsis clematidea. AB - A new codonopsine-related alkaloid and 13 known compounds were isolated from the aerial parts of Codonopsis clematidea (Campanulaceae). The structure of the new compound was elucidated by two-dimensional nuclear magnetic resonance and other spectral examinations. PMID- 18404332 TI - Cyclooxygenase-2 inhibitory effects and composition of the volatile oil from the dried roots of Lithospermum erythrorhizon. AB - The composition of the volatile oil from Lithospermi Radix, the dried roots of Lithospermum erythrorhizon (Boraginaceae), has been investigated by capillary GC and GC-MS. To investigate the anti-inflammatory activity of the oil, in-vitro inhibition of ovine cyclooxygenase-1 and 2 (COX-1 and COX-2) activity by the oil was studied. Fifty-four components of the oil were identified, representing 92.74% of the oil. The main components were 2-methylbutanoic acid (21.50%), 3 methylbutanoic acid (12.61%), 2-methylpropanoic acid (8.99%), methyl linoleate (8.76%), methyl oleate (6.27%), methyl palmitate (6.06%), and 2-methyl-2-butenoic acid (5.74%). Highly selective COX-2 inhibition was observed; at 50 microg/ml the oil inhibited 38.8% of COX-2 activity. PMID- 18404333 TI - Potential anthelmintic: D-psicose inhibits motility, growth and reproductive maturity of L1 larvae of Caenorhabditis elegans. AB - No anthelmintic sugars have yet been identified. Eight ketohexose stereoisomers (D- and L-forms of psicose, fructose, tagatose and sorbose), along with D galactose and D-glucose, were examined for potency against L1 stage Caenorhabditis elegans fed Escherichia coli. Of the sugars, D-psicose specifically inhibited the motility, growth and reproductive maturity of the L1 stage. D-Psicose probably interferes with the nematode nutrition. The present results suggest that D-psicose, one of the rare sugars, is a potential anthelmintic. PMID- 18404334 TI - Antioxidative activity of the flower of Torenia fournieri. AB - The edible flower of Torenia fournieri Linden ex E. Fourn was found to possess potent antioxidative activity in a rat brain homogenate model. Bioassay-guided isolation of the active compounds from a CH(2)Cl(2)-MeOH (1:1) extract led to the isolation of acteoside (1), luteolin-7-O-beta-glucoside (2), apigenin-7-O-alpha rhamnosyl-(1-->6)-beta-glucoside (3), and apigetrin (4). PMID- 18404335 TI - Chemical constituents from Oldenlandia corymbosa L. of Thai origin. AB - From the whole plants of Oldenlandia corymbosa of Thai origin, ten compounds have been isolated and elucidated as geniposide, 6 alpha-hydroxygeniposide, scandoside methyl ester (6 beta-hydroxygeniposide), asperulosidic acid, deacetylasperuloside, asperuloside, 10-O-benzoylscandoside methyl ester, 10-O-p hydroxybenzoylscandoside methyl ester, (+)-lyoniresinol-3 alpha-O-beta glucopyranoside, and rutin. Their structures were determined on the basis of spectroscopic data. PMID- 18404336 TI - Perilla frutescens var. frutescens in northern Laos. AB - Twenty-eight samples of mericarps of Perilla frutescens var. frutescens were collected through fieldwork performed in Phongsali and Xieng Khouang provinces in northern Laos. No perilla samples were collected from Savannakhet province in the south although more than 20 sites were investigated. Perilla plants are mostly grown mixed with dry-paddy rice by slash-and-burn cultivation in Laos. The most popular local name for perilla mericarps in the area was "Ma Nga Chan". Weight of 1,000 grains and hardness of the mericarps were measured, and all mericarps were found to be large (weight of 1,000 grains around 2 g) and soft (limit load weight under 300 g), which were preferred for culinary use in Laos. The composition of the essential oils obtained from the herbaceous plants raised from the mericarps was divided into five types, perillaketone, elemicine plus myristicine, shisofuran, piperitenon, and myristicine, and GC-MS analysis of these Laotian perilla samples showed that they were similar to those of corresponding types of known Japanese perilla strains. One of the shisofuran-type perilla contained large amounts of putative alpha-naginatene, which is likely to be an intermediate of the biosynthesis of naginataketone. The farmers' indifference to the oil type of the leaf seems to leave Laotian perilla as a good genetic resource for studies of the biosynthesis of oil compounds. PMID- 18404337 TI - Antibacterial activity of some medicinal plant extracts. AB - Antibacterial activity of hot aqueous and methanolic extracts prepared from six plants (Terminallia chebula, Terminallia bellerica, Phyllanthus emblica, Punica granatum, Lawsonia alba and Mikania micrantha) used in traditional folk medicines of India were screened against five pathogenic bacteria (Staphylococcus aureus MTCC 2940, Bacillus subtilis MTCC 441, Escherichia coli MTCC 739, Proteus vulgaris MTCC 426 and Enterobacter aerogenes MTCC 111). The highest antibacterial potentiality was exhibited by the methanolic leaf extract of T. chebula, followed by the aqueous fruit extract of T. bellerica. The leaf extract of T. chebula can be considered to be as equally potent as the most effective antibiotics, such as ciprofloxacin, gentamycin, kanamycin, ofloxacin and cephalexin. A sensitivity test performed with commonly used sensitivity test disks resulted in the appearance of multiple drug resistance phenotypes of the bacteria tested. A comparison of data in the inhibition zones of pathogenic bacteria showed that gentamycin, ofloxacin, kanamycin and tobramycin were effective against all of the bacterial strains tested. PMID- 18404339 TI - Antihyperglycaemic potential of the water-ethanol extract of Kalanchoe crenata (Crassulaceae). AB - Kalanchoe crenata is a vegetable widely used in Cameroon and largely efficient in the treatment of diabetes mellitus. The effect of the water-ethanol extract of this plant (WEKC) on blood glucose levels was investigated in fasting normal and diet-induced diabetic rats (MACAPOS 1) after a short- and medium-term treatment. Diabetes was induced by submitting Wistar rats to a hypercaloric sucrose diet over 4 months. Six hours after a single oral administration of WEKC, 135 and 200 mg kg(-1) body weight extracts significantly (P < 0.01) reduced the blood glucose levels both in normal and diabetic rats without real dose-dependent effect. During the medium-term treatment, 200 mg kg(-1) WEKC administered daily for 4 weeks significantly reduced blood glucose levels within week 1 (P < 0.05), with a maximum effect at week 4 (-52%, P < 0.01), while maintaining glycaemia within the normal range. All the WEKC-treated diabetic rats exhibited significant (P < 0.01) increase in insulin sensitivity index (K (ITT)) compared with the initial time and to the untreated diabetic animals. Animals treated for 4 weeks exhibited a slight resistance in body-weight gain and decrease in food and water intake. The WEKC activities on all parameters assessed were comparable with the glibenclamide effects. Qualitative phytochemical screening revealed that K. crenata contains terpenoids, tannins, polysaccharids, saponins, flavonoids and alkaloids. The data suggest that K. crenata might contain important chemical components that could induce significant improvement in glucose clearance and/or uptake and resistance to body-weight gain and insulin sensitivity, and could be a potent alternative or complementary therapeutic substance in the control of type 2 diabetes and other insulin-resistant conditions. PMID- 18404338 TI - Bioactive cyclobutane-containing alkaloids. AB - The present review describes research on novel natural cyclobutane-containing alkaloids and synthetic compounds isolated from terrestrial and marine species. More than 210 compounds have been confirmed to show antimicrobial, antibacterial, anticancer, and other activities. Structures, origins, biosynthesis, photodimerization, and biological activities of a selection of cyclobutane containing alkaloids and selected synthetic analogs of natural alkaloids are reviewed. PMID- 18404340 TI - Sedative effects of vapor inhalation of agarwood oil and spikenard extract and identification of their active components. AB - Agarwood oil and spikenard extract were examined for their sedative activity using a spontaneous vapor administration system. It was shown that inhalation of agarwood oil vapor sedated mice. The main volatile constituents of the oil were found to be benzylacetone [agarwood oil from a Hong Kong market (1)], or alpha gurjunene and (+)-calarene [agarwood oil made in Vietnam (2)]. A hexane extract of spikenard contained a lot of calarene, and its vapor inhalation had a sedative effect on mice. Individual principles benzylacetone, calarene, and alpha gurjunene were administered to mice, which reproduced the result of the corresponding oil or extract. However, the most effective dose of the compounds was lower than their original content in the oil and extract (benzylacetone 0.1%, calarene 0.17%, alpha-gurjunene 1.5%). PMID- 18404341 TI - Two new lignan glucosides from the fruit of Vitex cannabifolia. AB - Two new phenyldihydronaphthalene-type lignan glucosides, vitecannasides A (1) and B (2) were isolated from the fruit of Vitex cannabifolia Sieb. et. Zucc. (Verbenaceae) along with the six known lignan derivatives (3-8), four known iridoid glucosides (9-12), three known flavonoids (13-15), and one known phenylbutanone glucoside (16). Their chemical structures were determined on the basis of spectroscopic data as well as chemical evidence. The scavenging effect of 1, 2, and 11-16 on the stable free radical 1,1-diphenyl-2-picrylhydrazyl was examined. Compounds 1, 2, 14, and 15 exhibited stronger activity than that of L: cysteine. Especially, 14 and 15 showed more potent activity than that of alpha tocopherol. PMID- 18404343 TI - Two new glucuronide saponins, Achyranthosides G and H, from Achyranthes fauriei root. AB - Two oleanolic acid saponins named achyranthosides G (1) and H (2) were newly isolated from Achyranthes fauriei root as methyl esters in addition to methyl esters of achyranthosides A - F and five oleanolic acid glucuronides (chikusetsusaponins IVa, V, 28-deglucosyl chikusetsusaponin V, pseudoginsenoside RT(1), and oleanolic acid 3-O-beta-D-glucuronopyranoside) as well as oleanolic acid 28-O-beta-D-glucopyranoside, beta-ecdysterone, and polypodine B. Their structures were characterized as follows on the basis of the chemical and spectroscopic evidences. PMID- 18404344 TI - The relationship between 1-deoxynojirimycin content and alpha-glucosidase inhibitory activity in leaves of 276 mulberry cultivars (Morus spp.) in Kyoto, Japan. AB - The relationship between 1-deoxynojirimycin (DNJ) content and alpha-glucosidase inhibitory activity in mulberry (Morus) leaves is discussed. Mulberry leaves were collected from the Center for Bioresource Field Science, Kyoto Institute of Technology, Kyoto, Japan on 19 May, 9 July, and 9 August, 2003. Mulberry leaves were extracted with 75% ethanol. The inhibitory activity for rat intestinal crude enzyme was measured using maltose. The content of DNJ in the extracts was measured using HPLC. The mean DNJ content in the 0.04-0.06% range was high in collected samples. The inhibitory activities in July and August were higher (P < 0.01) than in May, and the activity in July was higher (P < 0.01) than in August. A strong correlation (r = 0.901, r (2) = 0.811, n = 15) existed between DNJ content and alpha-glucosidase inhibition in leaves of Morus bombycis harvested in July. Similarly, correlation coefficients of the other mulberry varieties in July were higher than they were in May or August. The inhibitory activity and the DNJ content of Morus latifolia in August were lower than for any other mulberry variety. These results show that the high inhibitory cultivars harvested in July, except for M. latifolia, are more suited to products that contain high DNJ contents. PMID- 18404342 TI - Antigenotoxic effect of nordihydroguaiaretic acid against chlormadinone acetate induced genotoxicity in mice bone-marrow cells. AB - Nordihydroguaiaretic acid (NDGA), a phenolic lignan, was tested for its antigenotoxic potential against chlormadinone acetate (CMA)-induced genotoxic damage in mice bone-marrow cells. Doses of about 22.50 mg/kg body weight of CMA were given along with 1, 5 and 10 mg/kg body weight of NDGA intraperitoneally. The treatment resulted in the reduction of sister chromatid exchanges and chromosomal aberrations induced by CMA, suggesting an antigenotoxic potential of NDGA. Earlier studies show that CMA generates reactive oxygen species, responsible for genotoxic damage. The free radical-scavenging property of NDGA is responsible for the reduction of genotoxic damage induced by CMA in mice bone marrow cells. PMID- 18404345 TI - Isolation and identification of antioxidants from Pedilanthus tithymaloides. AB - A bioassay-guided methodology utilizing 2,2-diphenyl-1-picrylhydrazyl (DPPH), the trolox equivalent antioxidant capacity (TEAC), and reducing power assays, as well as an assessment of scavenging properties against O (2) (.-) , H(2)O(2), HOCl, ROO., .NO, and ONOO(-) were used to find the main antioxidant principles of Pedilanthus tithymaloides (Euphorbiaceae), a shrub used in traditional Cuban medicine. The principles were identified as kaempferol 3-O-beta-D-glucopyranoside 6''-(3-hydroxy-3-methylglutarate), quercitrin, isoquercitrin, and scopoletin. The contents of total phenolics and flavonoids were found to be 76.0 +/- 4.8 mg of gallic acid equivalents/g extract and 9.8 +/- 0.4 mg of rutin equivalents/g extract, respectively. PMID- 18404346 TI - A new phenolic glucoside from an Uzbek medicinal plant, Origanum tyttanthum. AB - A new phenolic glucoside was isolated from the aerial parts of Origanum tyttanthum, an Uzbek medicinal plant, together with 12 known compounds. The structure of the new compound was elucidated as 4-O-beta-D-glucopyranosylbenzyl 3'-hydroxyl-4'-methoxybenzoate (1) based on the spectral and chemical evidence. PMID- 18404347 TI - Two new maltol glycosides and cyanogenic glycosides from Elsholtzia rugulosa Hemsl. AB - Two new maltol glycosides, maltol 6'-O-beta-D-apiofuranosyl-beta-D glucopyranoside and maltol 6'-O-(5-O-p-coumaroyl)-beta-D-apiofuranosyl-beta-D glucopyranoside, were isolated from Elsholtzia rugulosa Hemsl. along with 11 known compounds including prunasin and amygdalin. The structures were determined on the basis of spectroscopic and chemical evidence. This is the second example of isolation of cyanogenic glycosides from Lamiaceous plants. PMID- 18404348 TI - Anti-diabetic, anti-hyperlipidemic and spermatogenic effects of Amaranthus spinosus Linn. on streptozotocin-induced diabetic rats. AB - Anti-diabetic, anti-hyperlipidemic and spermatogenic effects were studies with methanolic extract of stem of Amaranthus spinosus Linn (Family: Amaranthaceae) in diabetic rats. In streptozotocin (STZ)-induced diabetic rats, it was observed that both the standard drug (Glibenclamide) and methanolic extract of Amaranthus spinosus Linn. significantly exhibited control of blood glucose level on a 15 day model. Further, the methanolic extract also showed significant anti hyperlipidemic and spermatogenic effects in STZ-induced diabetic rats. The methanolic extract has also accelerated the process of spermatogenesis by increasing the sperm count and accessory sex organ weights. The present investigation of the plant established some pharmacological evidence to support the folklore claim that it is used as an anti-diabetic. PMID- 18404350 TI - Coumarins from the roots of Ligusticum multivittatum. AB - Five new coumarins, (+)-peujaponisin (1) and multivittans A-D (2-5), were isolated from the roots of Ligusticum multivittatum Franch., and their structures were established by spectral means. PMID- 18404349 TI - Bioproduction of bile acids and the glycine conjugates by Penicillium fungus. AB - It has been found that the Penicillium endophytic filamentous fungus with the young stems of Scurrula atroprupurea (Loranthaceae) produces cholic acid, deoxycholic acid and the glycine conjugates. PMID- 18404351 TI - Assessment of the antibacterial activity of phenylethanoid glycosides from Phlomis lanceolata against multiple-drug-resistant strains of Staphylococcus aureus. AB - Three phenylethanoid glycosides, forsythoside B (1), phlinoside C (2) and verbascoside (3), were isolated from the methanol extract of the leaves of Phlomis lanceolata, an Iranian medicinal plant, by reversed-phase preparative high-performance liquid chromatography (HPLC), and the structures of these compounds were elucidated conclusively by ultraviolet (UV), mass spectrometry (MS) and a series of 1D and 2D nuclear magnetic resonance (NMR) analyses. The antibacterial properties of 1-3 against five multi-drug-resistant (MDR) strains of Staphylococcus aureus have been assessed by the rapid and robust microtitre plate-based serial dilution method. While compounds 1 and 3 showed considerable activities against all five strains, compound 2 was inactive at the test concentrations. PMID- 18404352 TI - Gastroprotective activity of Eriobotrya japonica seed extract on experimentally induced gastric lesions in rats. AB - The effect of Eriobotrya japonica seed extract (ESE) prepared with 70% ethanol on gastric mucosal injury was investigated. Six experimental models with different action mechanisms were used for the evaluation. Three concentrations of ESE were prepared for each model. ESE administration was initiated 14 days before induction of gastric mucosal injury, and its effect was investigated. ESE inhibited formation of gastric mucosal injury. PMID- 18404354 TI - Flavonoid glycosides and limonoids from Citrus molasses. AB - Molasses of tangerine orange (Citrus unshiu Markovich) is obtained as a waste product in the course of tangerine orange juice production. This molasses is expected to be a useful source of organic compounds such as flavonoids and limonoids. To elucidate a use for this molasses waste, we isolated and identified its organic constituents. Two new flavanonol glycosides were isolated from tangerine orange molasses, along with several flavonoids such as hesperidine, narirutin, eriodictyol, 3',4',5,6,7,8-hexamethoxy-3-O-beta-D glucopyranosyloxyflavone, and 3',4',5,6,7,8-hexamethoxy- 3-beta-D-[4-O-(3-hydroxy 3-methylglutaloyl)]-glucopyranosyloxyflavone, and limonoids such as limonin, nomilin, and cyclic peptide, citrusin III. The structures of the new flavanonol glycosides were determined as (2R,3R)-7-O-(6-O-alpha-L-rahmnopyranosyl-beta-D glucopyranosyl)-aromadendrin and 7-O-(6-O-alpha-L-rahmnopyranosyl-beta-D glucopyranosyl)-3,3',5,7-tetrahydroxy-4'-methoxyflavanone by means of spectral analyses using (1)H-NMR, (13)C-NMR, and 2D-NMR. Of these compounds, flavanone glycoside, hesperidin and narirutin were isolated as the main constituents. Thus, molasses is a promising source of flavonoid glycosides. PMID- 18404353 TI - DPPH radical-scavenging effect on some constituents from the aerial parts of Lippia triphylla. AB - Lippia triphylla (L'HER) O. KUNTZE: (Verbenaceae; common name, Lemon Verbena) is used in Peru as a spice and herb tea for the prevention of arteriosclerosis. From the aerial parts of this plant, 25 known compounds--3 phenylpropanoid glucosides, 7 flavonoids, 5 phenylethanoid glycosides, 5 lignans, 2 sesquiterpenoids, and 3 triterpenoids--were isolated, and their chemical structures were elucidated on the basis of physical and spectral data. Among them, 19 aromatic compounds were examined for their scavenging effect on the stable free radical 1,1-diphenyl-2 picrylhydrazyl--4 phenylethanoid glycosides and 5 lignans indicated a potent scavenging effect. Of note, the EC(50) values of two phenylethanoid glycosides reached almost thrice that of alpha-tocopherol. PMID- 18404355 TI - Comparative study on volatile components of Nardostachys rhizome. AB - Volatile components in 13 crude drug samples derived from Nardostachys chinensis or Nardostachys grandiflora were studied by solid phase micro extraction (SPME) GC and SPME-GC-MS. Twenty-three compounds accounting for 81.3 and 70.0% of volatile components in newly collected samples of two species were identified. beta-Maaliene, 9-aristolene, calarene and patchouli alcohol were identified as the major volatile constituents of N. chinensis, whereas aromadendrene, cube-11 ene, epi-alpha-selinene, spirojatamol and valeranone were identified as those of N. grandiflora. Using the peaks of beta-maaliene and 9-aristolene in GC profiles as the marker, two Nardostachys species were clearly distinguished among the samples examined. PMID- 18404356 TI - Authentication of Stephania tetrandra S. Moore (Fang Ji) and differentiation of its common adulterants using microscopy and HPLC analysis. AB - Stephania tetrandra S. Moore (Hang Fang Ji) is used in traditional Chinese medicine as a diuretic, an antiphlogistic, and an antirheumatic. The name "fang ji" is applied to at least four different genera of plants, including Aristolochia fangchi Y. C. Wu ex L. D. Chow and S. M. Hwang, Cocculus orbiculatus (L.) DC., Stephania tetrandra S. Moore, and Sinomenium acutum Rehder and E. H. Wilson. Due to similarity in the use of their common names, Stephania tetrandra S. Moore is often confused with Aristolochia fangchi Y. C. Wu ex L. D. Chow and S. M. Hwang, which has potentially dangerous consequences. To aid rapid and easy differentiation between the roots of these four species, so as to avoid possible contamination, detailed macroscopic and microscopic observations were made using stereo-and light-microscopy. The powdered samples were further analyzed using HPLC. PMID- 18404357 TI - Chemical constituents from the Paraguayan medicinal plant, Eupatorium macrocephalum Less. AB - This is the first phytochemical study of Eupatorium macrocephalum describing the isolation and identification of six triterpenes, one diterpene glycoside ester, six steroids, one flavonoid known as cirsilol, and six cinnamic acids derivatives. The structures of these known compounds were determined by spectral data analysis and comparison with literature values. PMID- 18404358 TI - Megastigmane and flavone glycosides from Strophioblachia fimbricalyx Boerl. AB - Three megastigmane glucosides, bridelionoside B, ampelopsisionoside and 3-hydroxy 5,6-epoxy-beta-ionol 9-O-beta-D-glucopyranoside, together with three flavone glycosides, apigenin 7-O-beta-D-glucopyranoside, isovitexin and spinosin, were isolated from the aerial portions of Strophioblachia fimbricalyx. The structure determinations were based on physical data and spectroscopic evidence. PMID- 18404359 TI - Rapid analysis of 56 pesticide residues in natural medicines by GC/MS with negative chemical ionization. AB - Many methods for determining pesticide residues in food have been reported. Although natural medicines should be confirmed to be as safe as food, few methods for determining pesticide residues in natural medicines have been reported. In this study, 56 pesticides were detected in natural medicines with a simple, rapid sample preparation method. This study indicates that the proposed method is useful for analyzing pesticides in natural medicines. PMID- 18404360 TI - Pain response of healthy workers following a functional capacity evaluation and implications for clinical interpretation. AB - BACKGROUND AND AIM: Functional Capacity Evaluations (FCEs) are used to quantify physical aspects of work capacity. Safety is a critical issue for clinical use of an FCE. Patients with Chronic Low Back Pain (CLBP) are known to report a temporary increase in pain following an FCE, but it is not known whether this increase is a normal pain response to FCE. It is currently unknown how healthy subjects respond to an FCE and whether this should be interpreted as a normal reaction after physical exercise. This study was performed to quantify the intensity, duration, location and nature of the pain response following an FCE in healthy subjects and to compare this pain response with the pain response of patients with CLBP from a previous study. METHODS: A total of 197 healthy working subjects between 20 and 60 years of age volunteered to participate in this study. All subjects performed a 12-item FCE. Pain response was measured by a self constructed Pain Response Questionnaire (PRQ). Descriptive statistics were used to describe the pain response following an FCE. Mann-Whitney and t-tests were performed to compare the data from this study with data of patients with CLBP from a previous study. RESULTS: About 82% of all subjects reported a pain response following the FCE. The intensity of the pain response after 24 h post FCE was a median of 3.0 on a numeric rating scale (0-10). About 78% of all pain was reducible to muscle soreness. Pain was most often reported in the upper legs (51%), the lower back (38%) the shoulders (37%) and upper arms (36%). Symptoms decreased to pre-FCE levels in a mean of 3 days. The pain response of 2 subjects (1%) lasted for 3 weeks. The intensity and duration of the pain response of healthy subjects was not significantly different from the response of patients with CLBP. CONCLUSION: Pain response of 99% of all subjects who reported a pain response was interpreted as normal. It was concluded that a pain response following an FCE can be expected in healthy subjects and that this pain response is a normal musculoskeletal reaction. The pain response of patients with CLBP resembles the pain response of healthy subjects. PMID- 18404361 TI - A prospective study of the effectiveness of early intervention with high-risk back-injured workers--a pilot study. AB - INTRODUCTION: It was postulated that workers, at the sub-acute stage after injury, respond differently to clinical and occupational interventions offered in a workers' compensation environment. Individual worker risk of disability, it was further believed, would influence the effectiveness of early intervention. The objective of the current pilot study was to evaluate return to work (RTW) outcomes following proactive, combined clinical, occupational and case management based interdisciplinary early intervention, provided in a workers' compensation environment 4-10 weeks of onset of back pain, to workers with medium and high risk for disability. METHODS: The project was a controlled study comparing conventional workers' compensation case management with integrated, interdisciplinary and multimodal early intervention (hereinafter referred to as "EI"). At baseline, risk status was determined by a validated Risk for Disability Questionnaire by Carragee et al. (Spine 5(1):24-35, 2005). Seventeen workers at high risk of protracted disability and 20 workers at moderate risk of disability received conventional case management, and 17 workers assessed at high risk of protracted disability and 18 workers at moderate risk of disability received the Early Intervention. RESULTS: At 3 months post back pain onset, no statistically significant differences were identified in RTW outcomes between conventional case management and the Early Intervention. However, by 6 months post back pain onset, workers at high risk of work disability who received the Early Intervention were significantly more likely to RTW than high risk workers who received conventional case management. In contrast, moderate risk workers continued to exhibit no statistically significant differences in RTW outcomes. CONCLUSION: Multimodal Early Intervention in the workers' compensation case management context is likely effective for workers with sub-acute back pain who are at high risk of occupational disability. The comprehensive Early Intervention is, however, likely redundant for workers who are not at high risk for disability and should not be applied indiscriminately. Further studies are required to determine longer-term Early Intervention outcomes, and to replicate the findings using a randomized control design. Also, with a larger sample size, it will be possible to determine predictors of occupational outcomes. PMID- 18404362 TI - Understanding systemic lupus erythematosus physiopathology in the light of primary immunodeficiencies. AB - INTRODUCTION: Associations between systemic lupus erythematosus (SLE) and primary immunodeficiencies (PIDs) were analyzed to gain insight into the physiopathology of SLE. Some PIDs have been consistently associated with SLE or lupus-like manifestations: (a) homozygous deficiencies of the early components of the classical complement pathway in the following decreasing order: in C1q, 93% of affected patients developed SLE; in C4, 75%; in C1r/s, 57%; and in C2, up to 25%; (b) female carriers of X-linked chronic granulomatous disease allele; and (c) IgA deficiency, present in around 5% of juvenile SLE. DISCUSSION: In the first two groups, disturbances of cellular waste-disposal have been proposed as the main mechanisms of pathogenesis. On the other hand and very interestingly, there are PIDs systematically associated with several autoimmune manifestations in which SLE has not been described, such as autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED), immunodysregulation polyendocrinopathy enteropathy X-linked (IPEX), and autoimmune lymphoproliferative syndrome (ALPS), suggesting that mechanisms considered as critical players for induction and maintenance of tolerance to autoantigens, such as (1) AIRE-mediated thymic negative selection of lymphocytes, (2) Foxp3+ regulatory T cell-mediated peripheral tolerance, and (3) deletion of auto-reactive lymphocytes by Fas-mediated apoptosis, could not be relevant in SLE physiopathology. The non-description of SLE and neither the most characteristic SLE clinical features among patients with agammaglobulinemia are also interesting observations, which reinforce the essential role of B lymphocytes and antibodies for SLE pathogenesis. CONCLUSION: Therefore, monogenic PIDs represent unique and not fully explored human models for unraveling components of the conundrum represented by the physiopathology of SLE, a prototypical polygenic disease. PMID- 18404363 TI - Effects of defendant sexual orientation on jurors' perceptions of child sexual assault. AB - We examined mock jurors' reactions to a sexual abuse case involving a male teacher and a 10-year-old child. Because gay men are sometimes stereotyped as child molesters, we portrayed defendant sexual orientation as either gay or straight and the victim as either a boy or girl. Jurors made more pro-prosecution decisions in cases involving a gay versus straight defendant, particularly when the victim was a boy. In boy-victim cases, jurors' emotional feelings of moral outrage toward the defendant mediated these effects. On average, women jurors were more pro-prosecution than were men. Results have implications for understanding social perceptions of cross- and same-gender child sexual abuse and juror decision making in child sexual assault cases perpetrated by homosexual and heterosexual men. PMID- 18404364 TI - Sexual risk behaviors in late middle age and older HIV seropositive adults. AB - Little is known about the sexual behaviors of older adults, although the prevalence of HIV/AIDS is rapidly increasing in this population. As part of a larger multi-site study examining secondary HIV prevention, we recruited from an HIV primary care clinic 210 sexually active HIV positive individuals aged 45 and over (125 men, 85 women) who had engaged in vaginal or anal sex within the past six months. Twenty percent of the participants reported inconsistent use of condoms and 33% had multiple sexual partners during the previous six months. Negative mood and perceived HIV stigma were associated with inconsistent condom use. In addition, multiple sex partners and higher level of education were related to inconsistent condom use during sex with partners of negative or unknown serostatus. These findings indicate that contrary to current beliefs, sexually active older adults, similar to younger ones, may be engaging in high risk transmission behaviors. PMID- 18404366 TI - John William Daly, 1933-2008. PMID- 18404365 TI - Hox-D genes expression in pediatric low-grade gliomas: real-time-PCR study. AB - HOX genes encode transcription factors, which play a key role in morphogenesis and cell differentiation during embryogenesis. Several observations indicate that a deregulated expression of these genes may result in tumor development and progression. Actually, several HOX genes are aberrantly expressed in many tumors and cell lines derived from them. Little is known about the expression of HOX genes in brain tumors. In the present work, we study the relative expression of HOX-D genes (D1, D3, D4, D8, D9, D10, D11, D12, D13) with real-time polymerase chain reaction in a group of 14 pediatric low-grade gliomas. We compare the HOX-D expression level of the 14 tumors with the average expression level of six non neoplastic human brain tissues. HOX-D1 and HOX-D12 resulted over-expressed in neoplastic samples with respect to non-neoplastic brain parenchyma. Conversely, HOX-D3 was expressed at a lower level in gliomas with respect to non-neoplastic brain. HOX-D4, HOX-D11, and HOX-D13 were never expressed. HOX-D8, HOX-D9, and HOX D10 were exceptionally expressed in non-neoplastic samples and irregularly expressed in tumors. The observation that all but three HOX-D genes studied are expressed with different pattern in neoplastic and non-neoplastic brain tissue may support the hypothesis that HOX-D genes play a role in the pathogenesis of pediatric low-grade gliomas. PMID- 18404367 TI - Glypican-3 regulates migration, adhesion and actin cytoskeleton organization in mammary tumor cells through Wnt signaling modulation. AB - Glypican-3 (GPC3) is a proteoglycan involved in migration, proliferation and cell survival modulation in several tissues. There are many reports demonstrating a downregulation of GPC3 expression in some human tumors, including mesothelioma, ovarian and breast cancer. Previously, we determined that GPC3 reexpression in the murine mammary adenocarcinoma LM3 cells induced an impairment of their in vivo invasive and metastatic capacities together with a higher susceptibility to in vitro apoptosis. Currently, the signaling mechanism of GPC3 is not clear. First, it was speculated that GPC3 regulates the insulin-like growth factor (IGF) signaling system. This hypothesis, however, has been strongly challenged. Recently, several reports indicated that at least in some cell types GPC3 serves as a selective regulator of Wnt signaling. Here we provide new data demonstrating that GPC3 regulates Wnt pathway in the metastatic adenocarcinoma mammary LM3 cell line. We found that GPC3 is able to inhibit canonical Wnt signals involved in cell proliferation and survival, as well as it is able to activate non canonical pathway, which directs cell morphology and migration. This is the first report indicating that breast tumor cell malignant properties can be reverted, at least in part, by GPC3 modulation of Wnt signaling. Our results are consistent with the potential role of GPC3 as a metastasis suppressor. PMID- 18404368 TI - The biological and clinical characteristics of breast carcinoma with mixed ductal and lobular morphology. AB - Although invasive ductal (IDC) and lobular (ILC) breast carcinomas are well characterised in the literature, the biological and clinical significance of mixed tumours with both ductal and lobular components has not been investigated. In the current study, we have examined a well-characterised series of breast carcinoma with a long term follow-up that comprised 140 mixed tumours, 2170 IDC and 380 pure ILC. RESULTS: Mixed tumours constituted 3.6% of all cases. The majority (59%) of the mixed tumours were grade 2 compared to 33% in IDC and 88% in ILC. Positive lymph nodes (LN) were found in 41% and definite vascular invasion (VI) in 26% of the cases. DCIS was detected in 123 (89%) and LCIS in 43 (31%) (both DCIS and LCIS were found in 39 cases). The majority of tumours were predominantly (>50 of tumour area) of ductal type (57%). When compared to pure IDC, mixed tumours showed an association with lower grade, ER positivity and lower frequency of development of distant metastases. When compared to pure ILC, mixed tumours showed an association with higher grade, positive LN metastasis, VI and development of regional metastasis. After adjustment for grade most of these differences were no longer apparent. There was an association between histologic type of carcinoma in LN metastasis and the predominant histologic type of the primary tumour. Mixed tumours showed metastatic patterns similar to that of ILC with frequent metastasis to bone. No clinically meaningful differences in survival were found between these mixed carcinomas and pure IDC or ILC of the breast or between mixed tumours with predominantly ductal or lobular phenotype. PMID- 18404370 TI - Transcriptional profiles of progestogen effects in the postmenopausal breast. AB - Estrogen plus progestin hormone therapy has been associated with increased breast proliferation, breast density, and breast cancer risk in postmenopausal women, beyond that seen with estrogen alone. The goal of this study was to evaluate progestogen effects on gene expression profiles in the breast contributing to this promotional effect. Twenty-five ovariectomized adult female cynomolgus monkeys were given the following treatments (expressed as equivalent doses for women) in a randomized crossover design: (1) placebo; (2) oral estradiol (E2, 1 mg/day); (3) E2 + micronized progesterone (P4, 200 mg/day); and (4) E2 + medroxyprogesterone acetate (MPA, 2.5 mg/day). Treatments were given for two months, and breast biopsies were taken after each treatment period. On microarray analysis E2 + MPA treatment resulted in a greater number of significantly regulated genes compared to E2 + P4 and E2 alone (P < 0.0001). Treatment with E2 alone induced modest effects on select genes related to epidermal growth factor receptor (EGFR) activity which were augmented by the addition of MPA but not P4, consistent with patterns of epithelial cell proliferation. Genes induced by E2 + MPA included the EGFR ligands EGF, TGFA, and AREG, and downstream targets such as STAT5A, STAT5B, SRC, EIF4EBP1, and MYC. Progestogens showed mixed antagonistic effects on E2-induced genes which tended to be greater for P4 than MPA. These findings suggest that a standard dose of oral E2 + MPA has a more pronounced effect on gene expression in the breast compared to E2 alone or E2 + P4 and that promotional effects of E2 + MPA may be mediated in part by increased EGFR activity. PMID- 18404369 TI - Promoter hypermethylation in sentinel lymph nodes as a marker for breast cancer recurrence. AB - PURPOSE: Promoter methylation of tumor suppressor genes in histologically negative sentinel lymph nodes (HNSN) of early stage breast cancer patients has not been extensively studied. This study evaluates the methylation frequency and pattern in HNSN to determine if detection of hypermethylation of one or more genes is associated with an increased recurrence risk in node negative breast cancer. EXPERIMENTAL DESIGN: In 1998, a prospective study of patients with early stage breast cancer and HNSN was initiated in order to correlate sentinel node analysis with clinical outcome. Nodal tissue was selected from 120 HNSN patients for methylation analysis in at least one and up to six sentinel nodes using a panel of nine genes. Corresponding primary breast tumors from 79 patients were also evaluated for hypermethylation. Methylation analysis was performed using nested Methylation Sensitive PCR (n-MSP). Logistical regression was used to evaluate the relationship between clinical recurrence and methylation status. RESULTS: Over a median follow-up of 79 months, 13 of the 120 patients had clinical recurrence. Hypermethylation of genes was frequently observed in HNSN, but there was no correlation of methylation pattern and clinical recurrence. However, increased frequency of gene methylation of the primary tumor correlated with clinical recurrence. CONCLUSIONS: Although hypermethylation of multiple genes occurs frequently in HNSN of breast cancer patients, it is not associated with breast cancer recurrence in the first 7 years of clinical follow-up. PMID- 18404371 TI - Convergence and divergence in the etiology of myelin impairment in psychiatric disorders and drug addiction. AB - Impairment of oligodendroglia (OL)-dependent myelination in the central nervous system (CNS) is a remarkable parallel recently identified in major psychiatric disorders and chronic drug abuse. Neuroimaging and neuropathological studies revealed myelin defects and microarray-profiling analysis demonstrated aberrant expression of myelin-related genes in schizophrenia (SZ), bipolar disorder (BD), major depressive disorder (MDD) and cocaine addiction. However, the etiology underlying myelin impairment in these clinically distinct subjects remains elusive. This article reviews myelin impairment in line with dopaminergic dysfunction, a prime neuropathophysiological trait shared in psychiatric disorders and drug abuse, as well as the genetic and epigenetic alterations associated with these diseases. The current findings support the hypothesis that aberrant dopamine (DA) action on OLs is a common pathologic mechanism for myelin impairment in the aforementioned mental morbidities, whereas inherited genetic variations that specifically affect OL development and myelinogenesis may further increase myelin vulnerability in psychiatric disorders. Importantly, OL defect is not only a pathological consequence but also a causative factor for dopaminergic dysfunction. Hence, myelin impairment is a key factor in the pathogenic loop of psychiatric diseases and drug addiction. PMID- 18404372 TI - Modulation of central nociceptive coding by acupoint stimulation. AB - It is universally accepted that acupuncture or acupoint stimulation can produce analgesic effect on patients with painful disorders. The past decades has seen remarkable progress in exploring the central mechanisms of acupuncture-induced pain relief, including the neurotransmitter release and expression of particular receptors and genes in the spinal cord and the brain stem regions. Development of new techniques makes it possible to record and image the brain network patterns underlying pain perception and modulation, and to investigate the role of higher level brain areas in mediating acupuncture analgesia. This review will present the current understanding of the neural network that is implicated in the modulation of pain by acupuncture. PMID- 18404374 TI - Roles of peripheral P2X and P2Y receptors in the development of melittin-induced nociception and hypersensitivity. AB - A recent report from our laboratory shows that subcutaneous (s.c.) injection of melittin could induce persistent spontaneous nociception (PSN) and primary thermal or mechanical hyperalgesia. However, the exact peripheral mechanisms underlying melittin-induced multiple pain-related behaviors remain unclear. In this study, behavioral tests combined with pharmacological manipulations were used to explore potential roles of local P2X and P2Y receptors in melittin induced inflammatory pain and hyperalgesia. Post-treatment of the primary injury site with s.c. injection of A-317491 (a potent P2X(3)/P2X(2/3) receptor antagonist) and Reactive Blue 2 (a potent P2Y receptor antagonist) could significantly suppress the development of melittin-evoked PSN and hypersensitivity (thermal and mechanical). Our control experiments demonstrated that local administration of either antagonist into the contralateral hindpaw produced no significant effect on any kind of pain-associated behaviors. Taken together, these data indicate that activation of P2X and P2Y receptors might be essential to the maintenance of melittin-induced primary thermal and mechanical hyperalgesia as well as on-going pain. PMID- 18404375 TI - Distinct responses of DREAM to electroacupuncture stimulation with different frequencies during physiological and inflammatory conditions in rats. AB - Our previous results indicated that dynorphin in the spinal dorsal horn mediates the analgesic effect of high frequency electroacupuncture stimulation (EAS). Here we report that the transcriptional repressor downstream regulatory element antagonist modulator (DREAM) of dynorphin precursor-preprodynorphin (PPD) may participate in this process. In normal rats, 100 Hz, but not 2 Hz EAS triggered the nuclear export and membrane translocation of DREAM concomitantly with the upregulation of PPD mRNA in the dorsal horn. In inflammatory rats, both 2 and 100 Hz EAS alleviated thermal and mechanical hypersensitivity and caused the nuclear export and membrane translocation of DREAM, but only 100 Hz EAS enhanced the mRNA level of PPD and DREAM. These results suggest the role of DREAM in the dorsal horn in the regulation of PPD gene expression by EAS is frequency dependent, and DREAM may exert different roles in different frequency EAS under physiological and inflammatory conditions. PMID- 18404373 TI - Therapeutic strategies for Parkinson's disease: the ancient meets the future- traditional Chinese herbal medicine, electroacupuncture, gene therapy and stem cells. AB - In China, it has been estimated that there are more than 2.0 million people suffering from Parkinson's disease, which is currently becoming one of the most common chronic neurodegenerative disorders during recent years. For many years, scientists have struggled to find new therapeutic approaches for this disease. Since 1994, our research group led by Drs. Ji-Sheng Han and Xiao-Min Wang of Neuroscience Research Institute, Peking University has developed several prospective treatment strategies for the disease. These studies cover the traditional Chinese medicine-herbal formula or acupuncture, and modern technologies such as gene therapy or stem cell replacement therapy, and have achieved some original results. It hopes that these data may be beneficial for the research development and for the future clinical utility for treatment of Parkinson's disease. PMID- 18404376 TI - Anionic selectivity sequence of the Cl(-)-H+ symporter in the synaptosomal preparation from rat brain cortex. AB - The Na(+)/H(+) exchanger has been the only unequivocally demonstrated H(+) transport mechanism in the synaptosomal preparation. We had previously suggested that a Cl(-)-H(+) symporter (in its acidifying mode) is involved in cytosolic pH regulation in the synaptosomal preparation. Supporting this suggestion, we now show that: (1) when synaptosomes are transferred from PSS to either gluconate or sulfate solutions, the Fura-2 ratio remains stable instead of increasing as it does in 50 mM K solution. This indicates that these anions do not promote a plasma membrane depolarization. (2) Based in the recovery rate from the cytosolic alkalinization, the anionic selectivity of the Cl(-)-H(+) symporter is NO(3)(-) > Br(-) > Cl(-) >> I(-) = isethionate = sulfate = methanesulfonate = gluconate. (3) PCMB 10 muM inhibits the gluconate-dependent alkalinization by 30 +/- 6%. (4) Neither Niflumic acid, 9AC, Bumetanide nor CCCP inhibits the recovery from the cytosolic alkalinization. PMID- 18404377 TI - Filamin A mutant lacking actin-binding domain restores mu opioid receptor regulation in melanoma cells. AB - We have previously reported that the protein filamin A (FLA) binds to the carboxyl tail of the mu opioid receptor (MOPr). Using human melanoma cells, which do not express filamin A, we showed that receptor down-regulation, functional desensitization and trafficking are deficient in the absence of FLA (Onoprishvili et al. Mol Pharmacol 64:1092-1100, 2003). Since FLA has a binding domain for actin and is a member of the family of actin cytoskeleton proteins, it is usually assumed that FLA functions via the actin cytoskeleton. We decided to test this hypothesis by preparing cDNA coding for mutant FLA lacking the actin binding domain (FLA-ABD) and expressing FLA-ABD in the human melanoma cell line M2 (M2 ABD cell line). We report here that this mutant is capable of restoring almost as well as full length FLA the down-regulation of the human MOPr. It is similarly very effective in restoring functional desensitization of MOPr, as assessed by the decrease in G-protein activation after chronic exposure of M2-ABD cells to the mu agonist DAMGO. We also found that A7 cells, expressing wild type FLA, exhibit rapid activation of the MAP kinases, ERK 1 and 2, by DAMGO, as shown by a rise in the level of phospho-ERK 1 and 2. This is followed by rapid dephosphorylation (inactivation), which reaches basal level between 30 and 60 min after DAMGO treatment. M2 cells show normal activation of ERK 1 and 2 in the presence of DAMGO, but very slow inactivation. The rapid rate of MAPK inactivation is partially restored by FLA-ABD. We conclude that some functions of FLA do not act via the actin cytoskeleton. It is likely that other functions, not studied here, may require functional binding of the MOPr-FLA complex to actin. PMID- 18404378 TI - Developmental regulation of TREM2 and DAP12 expression in the murine CNS: implications for Nasu-Hakola disease. AB - Trem2 is an orphan, DAP12 associated receptor constitutively expressed in vivo by subsets of microglia in the healthy adult murine CNS and in vitro by subsets of oligodendrocytes in neonatal mixed glial cultures. Loss of a functional Trem2 signaling pathway is the genetic cause of Nasu-Hakola disease. Whether the early onset cognitive dementia and myelin-pallor associated with this disorder are due to deficits in functional Trem2 signaling in microglia and/or oligodendrocytes is still being debated. Here, we find that Trem2/DAP12 expression is detected in embryonic day 14 CNS mRNA. Using dual immunohistochemistry/in situ hybridization, we find that both Trem2 and DAP12 expression always co-localized with markers of microglia/macrophages. However, Trem2/DAP12 positive microglia are found in very close apposition with CNP+ oligodendrocytes prior to myelination (post-natal day 1). In addition, CNS expression of TREM2 and DAP12 are not detected in PU.1KO which lack microglia and macrophages. Our data provide continuing support for Nasu-Hakola disease being identified as a cognitive disorder caused by a primary dysfunction of CNS microglia. PMID- 18404380 TI - Peer smoking, other peer attributes, and adolescent cigarette smoking: a social network analysis. AB - Peer attributes other than smoking have received little attention in the research on adolescent smoking, even though the developmental literature suggests the importance of multiple dimensions of adolescent friendships and peer relations. Social network analysis was used to measure the structure of peer relations (i.e., indicators of having friends, friendship quality, and status among peers) and peer smoking (i.e., friend and school smoking). We used three-level hierarchical growth models to examine the contribution of each time-varying peer variable to individual trajectories of smoking from age 11 to 17 while controlling for the other variables, and we tested interactions between the peer structure and peer smoking variables. Data were collected over five waves of assessment from a longitudinal sample of 6,579 students in three school districts. Findings suggest a greater complexity in the peer context of smoking than previously recognized. PMID- 18404381 TI - The Parents as Teachers program and school success: a replication and extension. AB - This study is a replication and extension of an earlier study, using a larger sample, a better measure of poverty status, and new longitudinal data. The study used path analysis to test hypothesized models of how the Parents as Teachers (PAT) program affects children's school readiness and subsequent third-grade achievement. Participants were 5,721 kindergarten children who were chosen to be representative of all children beginning public school in the state of Missouri in the fall of 1998-2000. These children were subsequently located in the state's third-grade test database 4-5 years later (82% of the original kindergarten sample). The causal models, which postulated both direct and indirect effects of PAT, were strongly supported by the data. EDITORS' STRATEGIC IMPLICATIONS: The findings add to the evidence that the PAT home visiting program holds promise as a primary prevention program. The authors demonstrate how parenting practices (including reading to children and enrolling them in preschool) promote both school readiness and subsequent academic achievement, but they also remind us of the pervasive effects of poverty. PMID- 18404379 TI - The protective effect of dexanabinol (HU-211) on nitric oxide and cysteine protease-mediated neuronal death in focal cerebral ischemia. AB - We hypothesized that dexanabinol can prevent neuronal death by protecting neuronal lysosomes from nitric oxide (NO)-mediated toxicity, and in turn, by suppressing the release of cathepsins during cerebral ischemia. Focal cerebral ischemia was induced in two sets of animals by permanent middle cerebral artery occlusion. The first set was used to monitor NO concentration and cathepsin activity, while the second was used for histological examination with hematoxylin and eosin, and TUNEL staining. In post-ischemic brain tissue, NO content and cathepsin B and L activity increased (p < 0.05). Dexanabinol treatment reduced NO concentration and cathepsin activity to the control level (p > 0.05). The number of eosinophilic and apoptotic neurons increased in the post-ischemic cerebral cortex (p < 0.05). However, dexanabinol treatment lowered both of these (p < 0.05). We conclude that dexanabinol might be a useful agent for the treatment of stroke patients. PMID- 18404383 TI - Transient asystole during endoscopic transsphenoidal surgery for acromegaly: an example of trigeminocardiac reflex. AB - Cardiac arrhythmias are rare during transsphenoidal surgery and is often secondary to stimulation of the trigeminal nerve endings that supply the nasal passages and cavernous sinus walls. Authors report a patient with Acromegaly, who developed transient asystole, during the dissection of the adenoma extending into the left cavernous sinus wall. In such cases, the use of prophylactic atropine may help to avoid such a complication. PMID- 18404382 TI - Osteogenesis Imperfecta: update on presentation and management. AB - Osteogenesis Imperfecta (OI) is a rare heritable condition characterized by bone fragility and reduced bone mass. Traditionally OI was classified into OI types I to IV and thought to be only due to a defect in the collagen gene, however through the discovery of the new types of OI-V to VII, breakthroughs have been made in understanding the pathophysiology of autosomal recessive OI and new genetic mutations, such as in CRTAP and P3H1 genes. OI can present at any age and be difficult to diagnose because of the wide phenotypic variation. Awareness of the new forms of OI, the differential diagnosis and the limitations of diagnostic tools, all help to correctly diagnose and manage a patient with OI. Cyclical intravenous pamidronate is now the standard of care for moderately to severely affected children with OI, given in combination with good orthopedic, physiotherapy and rehabilitation programs. The benefits and short term safety of cyclic bisphosphonates have been amply reported in the literature; however their long term effects are still under investigation. Newer more potent forms of bisphosphonates such as zoledronic acid have undergone and are still being subject to international multicentric drug trials and are beginning to replace pamidronate in some centers. PMID- 18404384 TI - Drugs and HPA axis. AB - This paper outlines the interferences of the most widely used drugs with hypothalamo-pituitary-adrenal function and the related laboratory parameters, with the purpose of providing practical help to clinicians during testing for hypo- or hypercortisolemic states. PMID- 18404386 TI - Male hypogonadism. AB - The hypothalamic-pituitary-gonadal (HPG) axis regulates the development, endocrine and reproductive function of the gonads throughout all phases of life. Male hypogonadism is defined an inadequate gonadal function, as manifested by deficiency in gametogenesis and/or secretion of gonadal hormones. In most cases, male hypogonadism is diagnosed through detailed history, physical examination and a few basic hormonal evaluations. In selected cases, however, additional tests are needed to define the aetiology and the extent of HPG axis dysfunction. These include semen analysis, pituitary imaging studies, genetic studies, bone densitometry, testicular ultrasonography, testicular biopsy and hormonal dynamic testing. The stimulation tests of the HPG are of particular importance in the differential diagnosis of congenital delayed puberty versus pre-pubertal hypogonadism in children. This review will focus on the methods, indications and limitations of endocrine testing in the characterisation and differential diagnosis of male hypogonadism at various ages. A practical hands-on guide on how to perform these tests is also provided. PMID- 18404385 TI - Systemic illness. AB - Systemic illnesses are associated with alterations in the hypothalamic-pituitary peripheral hormone axes, which represent part of the adaptive response to stressful events and may be influenced by type and severity of illness and/or pharmacological therapy. The pituitary gland responds to an acute stressful event with two secretory patterns: adrenocorticotropin (ACTH), prolactin (PRL) and growth hormone (GH) levels increase, while luteinizing hormone (LH), follicle stimulating hormone (FSH) and thyrotropin (TSH) levels may either decrease or remain unchanged, associated with a decreased activity of their target organ. In protracted critical illness, there is a uniformly reduced pulsatile secretion of ACTH, TSH, LH, PRL and GH, causing a reduction in serum levels of the respective target-hormones. These adaptations are initially protective; however, if inadequate or excessive they may be dangerous and may contribute to the high morbidity and mortality risk of these patients. There is no consensus regarding the type of approach, as well as the criteria to use to define pituitary axis function in critically ill patients. We here provide a critical approach to pituitary axis evaluation during systemic illness. PMID- 18404388 TI - Female hypogonadism: evaluation of the hypothalamic-pituitary-ovarian axis. AB - Female hypogonadism refers to deficient or abnormal function of the hypothalamic pituitary-ovarian axis that clinically presents with menstrual cycle disturbances. Female hypogonadism can be due to a congenital or acquired cause, and the defect can be at the level of the hypothalamus, pituitary or ovary. A careful history, physical exam and selected laboratory testing can often determine the locus of the defect and whether it results from a structural or hormonal problem. Laboratory testing generally relies on basal hormone levels; however, timing of blood sampling in relation to menses is important to interpretation of the data. PMID- 18404387 TI - Diagnosis of adult GH deficiency. AB - Based on previous consensus statements, it has been widely accepted that the diagnosis of adult growth hormone deficiency (GHD) must be shown biochemically by provocative tests of GH secretion; in fact, the measurement of IGF-I as well as of other markers was considered unable to distinguish between normal and GHD subjects. The Insulin Tolerance Test (ITT) was indicated as that of choice and severe GHD defined by a GH peak lower than 3 microg/l. It is now recognized that, although normal IGF-I levels do not rule out severe GHD, very low IGF-I levels in patients highly suspected for GHD (i.e. patients with childhood-onset severe GHD or with multiple hypopituitarism acquired in adulthood) can be considered as definite evidence for severe GHD. However, patients suspected for adult GHD with normal IGF-I levels must be investigated by provocative tests. ITT remains a test of reference but it should be recognized that other tests are as reliable as ITT. Glucagon as classical test and, particularly, new maximal tests such as GHRH in combination with arginine or GH secretagogues (GHS) (i.e. GHRP-6) have well defined cut-off limits, are reproducible, able to distinguish between normal and GHD subjects. Overweight and obesity have confounding effect on the interpretation of the GH response to provocative tests. In adults cut-off levels of GH response below which severe GHD is demonstrated must be appropriate to lean, overweight and obese subjects to avoid false positive diagnosis in obese adults and false negative diagnosis in lean GHD patients. PMID- 18404389 TI - Hyperprolactinemia. AB - In several respects prolactin is unique among anterior pituitary hormones. The primary regulation of prolactin secretion is mediated through hypothalamic inhibition, and the diagnosis of hyperprolactinemia can be established without the use of stimulation or suppression tests. Documenting the presence of hyperprolactinemia is not difficult-the challenge is in identifying the cause of the hormone hypersecretion. With immunoradiometric assays falsely low levels of prolactin are occasionally seen in patients with macroadenomas and very high serum prolactin (the hook effect). Macroprolactin should be suspected when a patient with hyperprolactinemia does not present with typical clinical symptoms, and all hyperprolactinemic sera should be screened for macroprolactin. With prolactinomas, prolactin levels generally parallel tumor size. Prolactin secreting macroadenomas are typically associated with levels that exceed 250 microg/l and may exceed 1,000 microg/l. Large non-functioning adenomas also lead to hyperprolactinemia but levels virtually never exceed 94 microg/l. Acquired and isolated prolactin deficiency is rare. PMID- 18404390 TI - Drugs and prolactin. AB - Medications commonly cause hyperprolactinemia and their use must be differentiated from pathologic causes. The most common medications to cause hyperprolactinemia are the antipsychotic agents, although some of the newer atypical antipsychotics do not do so. Other medications causing hyperprolactinemia include antidepressants, antihypertensive agents, and drugs which increase bowel motility. Often, the medication-induced hyperprolactinemia is symptomatic, causing galactorrhea, menstrual disturbance, and erectile dysfunction. In the individual patient, it is important differentiate hyperprolactinemia due to a medication from a structural lesion in the hypothalamic-pituitary area. This can be done by stopping the medication temporarily to determine if the prolactin (PRL) levels return to normal, switching to another medication in the same class which does not cause hyperprolactinemia (in consultation with the patient's physician and/or psychiatrist), or by performing an MRI or CT scan. If the hyperprolactinemia is symptomatic, management strategies include switching to an alternative medication which does not cause hyperprolactinemia, using estrogen/testosterone replacement, or cautiously adding a dopamine agonist. PMID- 18404391 TI - Does the type and severity of brain injury predict hypothalamo-pituitary dysfunction? Does post-traumatic hypopituitarism predict worse outcome? AB - Several studies have reported a close association between traumatic brain injury (TBI) and pituitary dysfunction, and expert panels have recently proposed recommendations for hormone assessment and replacement for pituitary insufficiency after TBI. Given the high incidence of TBI, identification of reliable predictors is of utmost importance in order to secure a cost-effective screening strategy. It has not yet been possible to identify early hormone alterations as a useful tool for the prediction of long-term post-traumatic hypopituitarism, whereas indicators of increased trauma severity have been reported as predictive in an increasing number of studies. Outcome studies have moreover indicated that post-traumatic hypopituitarism is of clinical significance, which may justify introduction of neuroendocrine screening in TBI. Much larger cohorts are, however, still needed for further evaluation and confirmation of reliable screening markers, and future studies should be designed to ensure a high diagnostic robustness for proper identification of reliable predictors, as the results may be highly dependent on diagnostic pitfalls. PMID- 18404392 TI - Social determinants, suboptimal health behavior, and morbidity in urban slum population: an Indian perspective. AB - Improving the health of urban residents, particularly those living in slum areas, requires an integrated approach. Appropriate interventions must be based on a well-grounded understanding of health determinants. Social factors are as important as physical factors in determining health status and suggest alternative interventions. Employment, stress, social exclusion, social support, substance use, nutrition, transport, and conditions during childhood are among the most important social determinants of health status identified by the International Center for Health and Society. This paper uses social determinants of health approach to understand morbidity outcomes for people residing in the slums of Surat City, India. To quantify suboptimal health behavior and identify the determinants of health status for this population survey data on household characteristics, health-seeking behavior, socioeconomic status, food and personal habits, social life, and physical activity has been used. After controlling for socioeconomic and demographic factors, logistic regression analysis reveals that social exclusion, stress, and lack of social support are significantly associated with morbidity. Thus, understanding of social determinants of health by policy makers is important as the health sector has a crucial role in addressing disparities in social determinants. PMID- 18404393 TI - Editorial. PMID- 18404394 TI - P2Y receptors and pain transmission. AB - It is widely accepted that the most important ATP receptors involved in pain transmission belong to the P2X(3) and P2X(2/3) subtypes, selectively expressed in small diameter dorsal root ganglion (DRG) neurons. However, several types of the metabotropic ATP (P2Y) receptors have also been found in primary afferent neurons; P2Y(1) and P2Y(2) receptors are typically expressed in small, nociceptive cells. Here we review the results available on the involvement of P2Y receptors in the modulation of pain transmission. PMID- 18404395 TI - P2X receptor-mediated purinergic sensory pathways to the spinal cord dorsal horn. AB - P2X receptors are expressed on different functional groups of primary afferent fibers. P2X receptor-mediated sensory inputs can be either innocuous or nociceptive, depending on which dorsal horn regions receive these inputs. We provide a brief review of P2X receptor-mediated purinergic sensory pathways to different regions in the dorsal horn. These P2X purinergic pathways are identified in normal animals, which provides insights into their physiological functions. Future studies on P2X purinergic pathways in animal models of pathological conditions may provide insights on how P2X receptors play a role in pathological pain states. PMID- 18404396 TI - The fate of P2Y-related orphan receptors: GPR80/99 and GPR91 are receptors of dicarboxylic acids. AB - Several orphan G protein-coupled receptors are structurally close to the family of P2Y nucleotide receptors: GPR80/99 and GPR91 are close to P2Y(1/2/4/6/11) receptors, whereas GPR87, H963 and GPR34 are close to P2Y(12/13/14). Over the years, several laboratories have attempted without success to identify the ligands of those receptors. In early 2004, two papers have been published: One claiming that GPR80/99 is an AMP receptor, called P2Y(15), and the other one showing that GPR80/99 is a receptor for alpha-ketoglutarate, while GPR91 is a succinate receptor. The accompanying paper by Qi et al. entirely supports that GPR80/99 is an alpha-ketoglutarate receptor and not an AMP receptor. The closeness of dicarboxylic acid and P2Y nucleotide receptors might be linked to the negative charges of both types of ligands and the involvement of conserved Arg residues in their neutralization. PMID- 18404397 TI - P2 receptors are involved in the mediation of motivation-related behavior. AB - The importance of purinergic signaling in the intact mesolimbic-mesocortical circuit of the brain of freely moving rats is reviewed. In the rat, an endogenous ADP/ATPergic tone reinforces the release of dopamine from the axon terminals in the nucleus accumbens as well as from the somatodendritic region of these neurons in the ventral tegmental area, as well as the release of glutamate, probably via P2Y(1) receptor stimulation. Similar mechanisms may regulate the release of glutamate in both areas of the brain. Dopamine and glutamate determine in concert the activity of the accumbal GABAergic, medium-size spiny neurons thought to act as an interface between the limbic cortex and the extrapyramidal motor system. These neurons project to the pallidal and mesencephalic areas, thereby mediating the behavioral reaction of the animal in response to a motivation-related stimulus. There is evidence that extracellular ADP/ATP promotes goal-directed behavior, e.g., intention and feeding, via dopamine, probably via P2Y(1) receptor stimulation. Accumbal P2 receptor-mediated glutamatergic mechanisms seem to counteract the dopaminergic effects on behavior. Furthermore, adaptive changes of motivation-related behavior, e.g., by chronic succession of starvation and feeding or by repeated amphetamine administration, are accompanied by changes in the expression of the P2Y(1) receptor, thought to modulate the sensitivity of the animal to respond to certain stimuli. PMID- 18404399 TI - Delineation of ligand binding and receptor signaling activities of purified P2Y receptors reconstituted with heterotrimeric G proteins. AB - P2Y receptors are G protein coupled receptors that respond to extracellular nucleotides to promote a multitude of signaling events. Our laboratory has purified several P2Y receptors with the goal of providing molecular insight into their: (1) ligand binding properties, (2) G protein signaling selectivities, and (3) regulation by RGS proteins and other signaling cohorts. The human P2Y(1) receptor and the human P2Y(12) receptor, both of which are intimately involved in ADP-mediated platelet aggregation, were purified to near homogeneity and studied in detail. After high-level expression from recombinant baculovirus infection of Sf9 insect cells, approximately 50% of the receptors were successfully extracted with digitonin. Purification of nearly homogeneous epitope-tagged P2Y receptor was achieved using metal-affinity chromatography followed by other traditional chromatographic steps. Yields of purified P2Y receptors range from 10 to 100 mug/l of infected cells. Once purified, the receptors were reconstituted in model lipid vesicles along with their cognate G proteins to assess receptor function. Agonist-promoted increases in steady-state GTPase assays demonstrated the functional activity of the reconstituted purified receptor. We have utilized this reconstitution system to assess the action of various nucleotide agonists and antagonists, the relative G protein selectivity, and the influence of other proteins, such as phospholipase C, on P2Y receptor-promoted signaling. Furthermore, we have identified the RGS expression profile of platelets and have begun to assess the action of these RGS proteins in a reconstituted P2Y receptor/G protein platelet model. PMID- 18404400 TI - Conserved lysine 79 is important for activity of ecto-nucleoside triphosphate diphosphohydrolase 3 (NTPDase3). AB - Cell membrane-bound ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are homooligomeric, with native quaternary structure required for maximal enzyme activity. In this study, we mutated lysine 79 in human ecto-nucleoside triphosphate diphosphohydrolase 3 (NTPDase3). The residue corresponding to lysine 79 in NTPDase3 is conserved in all known cell surface membrane NTPDases (NTPDase1, 2, 3, and 8), but not in the soluble, monomeric NTPDases (NTPDase5 and 6), or in the intracellular, two transmembrane NTPDases (NTPDase4 and 7). This conserved lysine is located between apyrase conserved region 1 (ACR1) and an invariant glycosylation site (N81), in a region previously hypothesized to be important for NTPDase3 oligomeric structure. This lysine residue was mutated to several different amino acids, and all mutants displayed substantially decreased nucleotidase activities. A basic amino acid at this position was found to be important for the increase of nucleotidase activity observed after treatment with the lectin, concanavalin A. After solubilization with Triton X-100, mutants showed little or no decrease in activity, unlike the wild-type enzyme, suggesting that the lysine at this position may be important for maintaining proper folding and for stabilizing the quaternary structure. However, mutation at this site did not result in global changes in tertiary or quaternary structure as measured by Cibacron blue binding, chemical cross linking, and native gel electrophoretic analysis, leaving open the possibility of other mechanisms by which mutation of this conserved lysine residue might decrease enzyme activity. PMID- 18404398 TI - Regulation of neuronal ion channels via P2Y receptors. AB - Within the last 15 years, at least 8 different G protein-coupled P2Y receptors have been characterized. These mediate slow metabotropic effects of nucleotides in neurons as well as non-neural cells, as opposed to the fast ionotropic effects which are mediated by P2X receptors. One class of effector systems regulated by various G protein-coupled receptors are voltage-gated and ligand-gated ion channels. This review summarizes the current knowledge about the modulation of such neuronal ion channels via P2Y receptors. The regulated proteins include voltage-gated Ca(2+) and K(+) channels, as well as N-methyl-D: -aspartate, vanilloid, and P2X receptors, and the regulating entities include most of the known P2Y receptor subtypes. The functional consequences of the modulation of ion channels by nucleotides acting at pre- or postsynaptic P2Y receptors are changes in the strength of synaptic transmission. Accordingly, ATP and related nucleotides may act not only as fast transmitters (via P2X receptors) in the nervous system, but also as neuromodulators (via P2Y receptors). Hence, nucleotides are as universal transmitters as, for instance, acetylcholine, glutamate, or gamma-aminobutyric acid. PMID- 18404401 TI - Coronary artery reperfusion: The ADP receptor P2Y(1) mediates early reactive hyperemia in vivo in pigs. AB - The physiological mechanisms that regulate reactive hyperemia are not fully understood. We postulated that the endothelial P2Y(1) receptor that release vasodilatory factors in response to ADP might play a vital role in the regulation of coronary flow. Intracoronary flow was measured with a Doppler flow-wire in a porcine model. 2-MeSADP (10(-5) M), ATP (10(-4) M) or UTP (10(-4) M) alone or as co-infusion with a selective P2Y(1) receptor blocker, MRS 2179 (10(-3) M) was locally delivered through the tip of a coronary angioplasty balloon. In separate pigs the coronary artery was occluded with the balloon for 10 min. During the first and tenth minutes of coronary ischemia, 2.5 ml of MRS 2179 (10(-3) M) was delivered distal to the occlusion in 8 pigs, 10 pigs were used as controls. MRS 2179 fully inhibited the 2-MeSADP-mediated coronary flow increase (P < 0.05) with no effect on UTP, indicating selective P2Y(1) inhibition. ATP-mediated flow increase was significantly inhibited by MRS 2179. During reactive hyperemia following coronary occlusion, flow increased by nearly sevenfold. MRS 2179, however, reduced the post-ischemic hyperemia by a mean of 46% during the period 1 2.5 min following balloon deflation (P < 0.05), which corresponds to peak velocity flow during reperfusion. In conclusion, MRS 2179, a selective P2Y(1) receptor blocker, significantly reduces the increased coronary flow caused both by 2-MeSADP and reactive hyperemia in coronary arteries. Thus, ADP acting on the endothelial P2Y(1) receptor may play a major role in coronary flow during post ischemic hyperemia. PMID- 18404402 TI - GPR80/99, proposed to be the P2Y(15) receptor activated by adenosine and AMP, is not a P2Y receptor. AB - The orphan receptor GPR80 (also called GPR99) was recently reported to be the P2Y(15) receptor activated by AMP and adenosine and coupled to increases in cyclic AMP accumulation and intracellular Ca(2+) mobilization (Inbe et al. J Biol Chem 2004; 279: 19790-9). However, the cell line (HEK293) used to carry out those studies endogenously expresses A(2A) and A(2B) adenosine receptors as well as multiple P2Y receptors, which complicates the analysis of a potential P2Y receptor. To determine unambiguously whether GPR80 is a P2Y receptor subtype, HA tagged GPR80 was either stably expressed in CHO cells or transiently expressed in COS-7 and HEK293 cells, and cell surface expression was verified by radioimmunoassay (RIA). COS-7 cells overexpressing GPR80 showed a consistent twofold increase in basal inositol phosphate accumulation. However, neither adenosine nor AMP was capable of promoting accumulation of either cyclic AMP or inositol phosphates in any of the three GPR80-expressing cells. A recent paper (He et al. Nature 2004; 429: 188-93) reported that GPR80 is a Gq-coupled receptor activated by the citric acid cycle intermediate, alpha-ketoglutarate. Consistent with this report, alpha-ketoglutarate promoted inositol phosphate accumulation in CHO and HEK293 cells expressing GPR80, and pretreatment of GPR80-expressing COS-7 cells with glutamate dehydrogenase, which converts alpha-ketoglutarate to glutamate, decreased basal levels of inositol phosphates. Taken together, these data demonstrate that GPR80 is not activated by adenosine, AMP or other nucleotides, but instead is activated by alpha-ketoglutarate. Therefore, GPR80 is not a new member of the P2Y receptor family. PMID- 18404403 TI - P2Y(1) receptor modulation of endogenous ion channel function in Xenopus oocytes: Involvement of transmembrane domains. AB - Agonist activation of the hP2Y(1) receptor expressed in Xenopus oocytes stimulated an endogenous voltage-gated ion channel, previously identified as the transient inward (T(in)) channel. When human P2Y(1) (hP2Y(1)) and skate P2Y (sP2Y) receptors were expressed in Xenopus oocytes, time-to-peak values (a measure of the response to membrane hyperpolarization) of the T(in) channel were significantly reduced compared to oocytes expressing the hB(1)-bradykinin receptor or the rat M(1)-muscarinic (rM(1)) receptor. Differences in activation were also observed in the T(in) currents elicited by various P2Y receptor subtypes. The time-to-peak values of the T(in) channel in oocytes expressing the hP2Y(4), hP2Y(11), or hB(1)-bradykinin receptors were similar, whereas the channel had significantly shorter time-to-peak values in oocytes expressing either the hP2Y(1) or sP2Y receptor. Amino acid substitutions at His-132, located in the third transmembrane domain (TM3) of the hP2Y(1) receptor, delayed the onset of channel opening, but not the kinetics of the activation process. In addition, Zn(2+) sensitivity was also dependent on the subtype of P2Y receptor expressed. Replacement of His-132 in the hP2Y(1) receptor with either Ala or Phe increased Zn(2+) sensitivity of the T(in) current. In contrast, truncation of the C-terminal region of the hP2Y(1) receptor had no affect on activation or Zn(2+) sensitivity of the T(in) channel. These results suggested that TM3 in the hP2Y(1) receptor was involved in modulating ion channel function and blocker pharmacology of the T(in) channel. PMID- 18404404 TI - Immunolocalisation of P2Y receptors in the rat eye. AB - Nucleotides present an important role in ocular physiology which has been demonstrated by recent works that indicate their involvement in many ocular processes. P2Y are important among P2 receptors since they can control tear production, corneal wound healing, aqueous humour dynamics and retinal physiology. Commercial antibodies have allowed us to investigate the distribution of P2Y receptors in the cornea, anterior and posterior chamber of the eye and retina. The P2Y(1) receptor was present mainly in cornea, ciliary processes, and trabecular meshwork. The P2Y(2) receptors were present in cornea, ciliary processes and retinal pigmented epithelium. P2Y(4) was present in cornea, ciliary processes, photoreceptors, outer plexiform layer and ganglion cell layer. The P2Y(6) presented almost an identical distribution as the P2Y(4) receptor. The P2Y(11) was also detectable in the retinal pigmented epithelium. The detailed distribution of the receptors clearly supports the recent findings indicating the relevant role of nucleotides in the ocular function. PMID- 18404405 TI - Influences of calcium deficiency and cerium on the conversion efficiency of light energy of spinach. AB - Chloroplast absorbs light energy and transforms it into electron energy, and then converts it into active chemical energy and stable chemical energy. In the present paper, we investigated the effects of Ce(3+), which has the most significant catalytic effects and similar characteristics with Ca(2+), on light energy conversion of spinach chloroplasts under Ca(2+)-deficient stress. The results illuminated that the Hill reaction activity, electron flow both photosystems and photophosphorylation rate of spinach chloroplasts reduced significantly under Ca(2+)-deficient condition, and activities of Mg(2+)-ATPase and Ca(2+)-ATPase on the thylakoid membrane were severely inhibited. Meanwhile, the activity of Rubisco, which is the key enzyme of photosynthetic carbon assimilation, was also prohibited. However, Ce(3+) decreased the inhibition of calcium deprivation the electron transport rate, the oxygen evolution rate, the cyclic and noncyclic photophosphorylation, the activities of Mg(2+)-ATPase, Ca(2+)-ATPase and Rubisco of spinach chloroplasts. All above implied that Ca(2+) depletion could disturb light energy conversion of chloroplasts strongly, which could be reversed by Ce(3+). PMID- 18404406 TI - Differences between women and men in serial HIV prevalence and incidence trends. AB - To measure trends in HIV incidence and serial prevalence by sex in a intravenous drug users (IDUs) and heterosexuals (HT) cohort recruited in a counselling centre in Valencia (1988-2005). Serial prevalence and incidence rates were calculated and modelled by logistic and Poisson regression respectively. 5948 IDUs and 13343 HT were recruited. Prevalence was higher among female IDUs (46% vs. 41%), and female HT (4.1% vs. 2%). For IDUs, an interaction (P = 0.005) between sex and calendar was detected. Age-adjusted prevalence showed faster yearly decline in men (OR = 0.87 95%CI: 0.85-0.88) than in women (OR = 0.91 95%CI: 0.88-0.93). Incidence was higher in female IDUs (9.79% p-y) than in men, (5.38% p-y) with an annual decrease for both of 11%. HIV incidence was higher in female HT (0.62% p y) compared to men 0.23% p-y with a 21% yearly decline. Gender differences in HIV prevalence and incidence trends have been detected. Women showed an increased vulnerability to infection in a country whose HIV epidemic has been largely driven by IDUs. PMID- 18404407 TI - Variance estimation of allele-based odds ratio in the absence of Hardy-Weinberg equilibrium. AB - In gene-disease association studies, deviation from Hardy-Weinberg equilibrium in controls may cause bias in estimating the allele-based estimates of genetic effects. An approach to adjust the variance of allele-based odds ratio for Hardy Weinberg equilibrium deviation is proposed. Such adjustments have been introduced for estimating relative risks of genotype contrasts and differences in allele frequency; however, an adjustment of odds ratios for allele frequencies still does not exist. The approach was based on the delta method in combination with the Woolf's logit interval method and the disequilibrium coefficient. The proposed variance adjustment provided better power than the unadjusted one to detect significant estimates of odds ratio and it improved the variance estimation. PMID- 18404408 TI - Population dynamics of phytophagous and predaceous mites on coffee in Brazil, with emphasis on Brevipalpus phoenicis (Acari: Tenuipalpidae). AB - The objective of this work was to study the population dynamics of Brevipalpus phoenicis (Geijskes) and predaceous mites (Phytoseiidae and Stigmaeidae), as well as the interactions among these mite species, in a coffee plantation in the State of Sao Paulo, Brazil. Tydeids were also evaluated because of the high frequency of these mites on coffee plants. Samples of leaves, branches and fruits were taken fortnightly, from April 2001 to June 2003, from plants randomly chosen in the coffee plantation. B. phoenicis mites were found on leaves in higher number during the drier periods of the year. Among the predaceous mites, Euseius citrifolius Denmark & Muma and Euseius concordis (Chant) were the most frequent species on the surface of leaves. During the evaluations, 72,534 domatia were cut and opened to remove the mites, from 6,360 leaves examined. Zetzellia malvinae Matioli, Ueckermann & Oliveira was the most frequent predator species found inside domatia. Significant correlations were observed between population dynamics of, among others, E. concordis and B. phoenicis, Z. malvinae and B. phoenicis, and Z. malvinae and E. concordis. Significant correlations were also verified between the number of domatia and the population densities of B. phoenicis, E. concordis, Lorryia sp. and Z. malvinae. Interactions between predator-prey and predator-predator on coffee plants are discussed. The influence of the meteorological factors temperature and precipitation on the most frequent mite species is also discussed. PMID- 18404409 TI - Determination of the effects of temporal change in urban and agricultural land uses as seen in the example of the town of Akhisar, using remote sensing techniques. AB - Today, as a result of erratic and unplanned urbanization, towns are rapidly becoming a mass of concrete and town-dwellers are suffocated by their busy and stressful professional lives. They feel a need for places where they can find breathing-space in their free time. Green areas within towns are important spaces where townspeople are able to carry out recreational activities. These places form a link between townspeople and nature. The importance of urban green areas is increasing with every passing day due to their social, psychological, ecological, physical and economic functions and their impact on the quality of towns. In this study it has been attempted to demonstrate the pressures of urban development on agricultural land by determining the changing land use situation over the years in the district of Akhisar. In this research, an aerial photograph from year 1939 and satellite images of the town from the years 2000 and 2007 were used. Land use changes in the region were determined spatially. As a result of this study, which aims to determine in which direction urbanization is progressing in the district, the importance of town planning emerges. This study will be informative for the local authorities in their future town planning projects. With its flat and almost flat fertile arable land, the district of Akhisar occupies an important position within the province of Manisa. From the point of view of olive production the region is one of Turkey's important centres. Fifty-five percent of the olive production in the province of Manisa is realized in Akhisar. However, the results of the present study show that while agricultural areas comprised 2.5805 km(2) in 1939, these had diminished to 1.5146 km(2) in the year 2000 and had diminished to 1.0762 km(2) in the year 2007 and residential area (dense) 0.449 km(2) occupied in 1939, in the year 2000 this had risen to 1.9472 and 2.3238 km(2) in the year 2007. This planless urbanization in the study area has led to great losses of farmland. PMID- 18404410 TI - Environmental impact of herbicide regimes used with genetically modified herbicide-resistant maize. AB - With the potential advent of genetically modified herbicide-resistant (GMHR) crops in the European Union, changes in patterns of herbicide use are predicted. Broad-spectrum, non-selective herbicides used with GMHR crops are expected to substitute for a set of currently used herbicides, which might alter the agro environmental footprint from crop production. To test this hypothesis, the environmental impact of various herbicide regimes currently used with non-GMHR maize in Belgium was calculated and compared with that of possible herbicide regimes applied in GMHR maize. Impacts on human health and the environment were calculated through the pesticide occupational and environmental risk (POCER) indicator. Results showed that the environmental impact of herbicide regimes solely relying on the active ingredients glyphosate (GLY) or glufosinate-ammonium (GLU) is lower than that of herbicide regimes applied in non-GMHR maize. Due to the lower potential of GLY and GLU to contaminate ground water and their lower acute toxicity to aquatic organisms, the POCER exceedence factor values for the environment were reduced approximately by a sixth when GLY or GLU is used alone. However, the environmental impact of novel herbicide regimes tested may be underestimated due to the assumption that active ingredients used with GMHR maize would be used alone. Data retrieved from literature suggest that weed control efficacy is increased and resistance development delayed when GLY or GLU is used together with other herbicides in the GMHR system. Due to the partial instead of complete replacement of currently used herbicide regimes, the beneficial environmental impact of novel herbicide regimes might sometimes be reduced or counterbalanced. Despite the high weed control efficacy provided by the biotechnology-based weed management strategy, neither indirect harmful effects on farmland biodiversity through losses in food resources and shelter, nor shifts in weed communities have been demonstrated in GMHR maize yet. However, with the increasing adoption rate of GMHR maize and their associated novel herbicide regimes, this situation is expected to change in the short-term. PMID- 18404411 TI - 'Non-neuronopathic' Gaucher disease reconsidered. Prevalence of neurological manifestations in a Dutch cohort of type I Gaucher disease patients and a systematic review of the literature. AB - Gaucher disease is a lysosomal storage disorder, which is classically divided into three types. Type I Gaucher disease is differentiated from types II and III disease by the absence of nervous system involvement. However, an increasing number of reports has emerged on neurological manifestations in patients with type I Gaucher disease. Whether a strict division in three different phenotypes is still valid has been the subject of debate. The main objective of this study was to provide scientific arguments whether a distinction between type I (non neuronopathic) and types II and III (neuronopathic) Gaucher disease should be maintained. We investigated retrospectively a large Dutch cohort of type I Gaucher disease patients for the prevalence of neurological manifestations and provide an overview of the literature on this topic. A diagnosis of a neurological disease was made 34 times in 75 patients. Forty-five patients reported at least one neurological symptom during the median follow-up time of 11 years. The literature search revealed 86 studies in which type I Gaucher disease patients or carriers of a glucocerebrosidase mutation were described with a neurological disease or a condition which is known to be associated with neurological disease. In conclusion, the term non-neuronopathic Gaucher disease does not seem to be an appropriate characterization of type I Gaucher disease. However, the neurological signs and symptoms in type I Gaucher disease are of a totally different kind from and, in the majority of cases, of much less severity than the signs and symptoms associated with types II and III disease Therefore, type I disease should be classified as a separate phenotype. PMID- 18404412 TI - Membrane translocation of glutaric acid and its derivatives. AB - The neurodegenerative disorder glutaric aciduria type I (GA I) is characterized by increased levels of cytotoxic metabolites such as glutaric acid (GA) and 3 hydroxyglutaric (3OHGA). The present report summarizes recent investigations providing insights into mechanisms of intra- and intercellular translocation of these metabolites. Initiated by microarray analyses in a mouse model of GA I, the sodium-dependent dicarboxylate cotransporter 3 (NaC3) was the first molecule identified to mediate the translocation of GA and 3OHGA with high and low affinity, respectively. More recently, organic anion transporters (OAT) 1 and 4 have been reported to be high-affinity transporters for GA and 3OHGA as well as D 2- and L-2-hydroxyglutaric acid (D2OHGA, L2OHGA). The concerted action of NaC3 and OATs may be important for the directed uptake and excretion of GA, 3OHGA, D2OHGA and L2OHGA in kidney proximal tubule cells. In addition, experimental data on cultured neuronal and glial cells isolated from mouse brain demonstrated that GA rather than 3OHGA may competitively inhibit the anaplerotic supply of tricarboxylic acid cycle intermediates from astrocytes to neurons. The identification of GA and GA derivative transporters may represent targets for new approaches to treat patients with GA I and related disorders. PMID- 18404413 TI - Purinergic signalling in the immune system. A brief update. PMID- 18404414 TI - Chemotactic activity of extracellular nucleotideson human immune cells. AB - Purinergic P2 receptors are a class of plasma membrane receptors that are express in many tissues and are ligated by extracellular nucleotides [such as adenosine triphosphate (ATP), adenosine diphosphate (ADP), uridine 5'-triphosphate (UTP) and uridine 5'-diphosphate (UDP)], which are released as a consequence of cell damage, cell stress, bacterial infection or other noxious stimuli. According to the molecular structure, P2 receptors are divided into two subfamilies: P2X and P2Y receptors. The P2X receptors are ligand-gated channels, whereas P2Y receptors are G-protein-coupled seven-membrane-spanning receptors. Several studies indicate that nucleotides play an important role in immune response modulation through their action on multiple cell types, including monocytes, mast cells, dendritic cells, neutrophils, and eosinophils. Recent work by our group and others identified extracellular nucleotides as chemotaxins for various human immune cells, including eosinophils, neutrophils and dendritic cells. In this review, we summarise recent findings in this field and put forward a hypothesis on the role of P2 receptors in the early recruitment of human immune cells to the site of inflammation. PMID- 18404415 TI - P1 receptors and cytokine secretion. AB - Evidence has accumulated in the last three decades to suggest tissue protection and regeneration by adenosine in multiple different cell types. Adenosine produced in hypoxic or inflamed environments reduces tissue injury and promotes repair by receptor-mediated mechanisms. Among other actions, regulation of cytokine production and secretion by immune cells, astrocytes and microglia (the brain immunocytes) has emerged as a main mechanism at the basis of adenosine effects in diseases characterized by a marked inflammatory component. Many recent studies have highlighted that signalling through A(1) and A(2A) adenosine receptors can powerfully prevent the release of pro-inflammatory cytokines, thus inhibiting inflammation and reperfusion injury. However, the activation of adenosine receptors is not invariably protective of tissues, as signalling through the A(2B) adenosine receptor has been linked to pro-inflammatory actions which are, at least in part, mediated by increased release of pro-inflammatory cytokines from epithelial cells, astrocytes and fibroblasts. Here, we discuss the multiple actions of P1 receptors on cytokine secretion, by analyzing, in particular, the role of the various adenosine receptor subtypes, the complex reciprocal interplay between the adenosine and the cytokine systems, their pathophysiological significance and the potential of adenosine receptor ligands as new anti-inflammatory agents. PMID- 18404416 TI - P2 purinergic receptor modulation of cytokine production. AB - Cytokines serve important functions in controlling host immunity. Cells involved in the synthesis of these polypeptide mediators have evolved highly regulated processes to ensure that production is carefully balanced. In inflammatory and immune disorders, however, mis-regulation of the production and/or activity of cytokines is recognized as a major contributor to the disease process, and therapeutics that target individual cytokines are providing very effective treatment options in the clinic. Leukocytes are the principle producers of a number of key cytokines, and these cells also express numerous members of the purinergic P2 receptor family. Studies in several cellular systems have provided evidence that P2 receptor modulation can affect cytokine production, and mechanistic features of this regulation have emerged. This review highlights three separate examples corresponding to (1) P2Y6 receptor mediated impact on interleukin (IL)-8 production, (2) P2Y11 receptor-mediated affects on IL-12/23 output, and (3) P2X7 receptor mediated IL-1beta posttranslational processing. These examples demonstrate important roles of purinergic receptors in the modulation of cytokine production. Extension of these cellular observations to in vivo situations may lead to new therapeutic strategies for treating cytokine mediated diseases. PMID- 18404417 TI - Nucleotide receptor signalling and the generation of reactive oxygen species. AB - Elevated levels of extracellular nucleotides are present at sites of inflammation, platelet degranulation and cellular damage or lysis. These extracellular nucleotides can lead to the activation of purinergic (nucleotide) receptors on various leukocytes, including monocytes, macrophages, eosinophils, and neutrophils. In turn, nucleotide receptor activation has been linked to increased cellular production and release of multiple inflammatory mediators, including superoxide anion, nitric oxide and other reactive oxygen species (ROS). In the present review, we will summarize the evidence that extracellular nucleotides can facilitate the generation of multiple ROS by leukocytes. In addition, we will discuss several potential mechanisms by which nucleotide enhanced ROS production may occur. Delineation of these mechanisms is important for understanding the processes associated with nucleotide-induced antimicrobial activities, cell signalling, apoptosis, and pathology. PMID- 18404418 TI - P2 receptors in macrophage fusion and osteoclast formation. AB - Cells of the mononuclear phagocyte lineage fuse to form multinucleated giant cells and osteoclasts. Several lines of evidence suggest that P2 receptors, in particular P2X7, are involved in this process, although P2X7 is not absolutely required for fusion because P2X7-null mice form multinucleated osteoclasts. Extracellular ATP may be an important regulator of macrophage fusion. PMID- 18404419 TI - Activation of transcription factors by extracellular nucleotides in immune and related cell types. AB - Extracellular nucleotides, acting through P2 receptors, can regulate gene expression via intracellular signaling pathways that control the activity of transcription factors. Relatively little is known about the activation of transcription factors by nucleotides in immune cells. The NF-kappaB family of transcription factors is critical for many immune and inflammatory responses. Nucleotides released from damaged or stressed cells can act alone through certain P2 receptors to alter NF-kappaB activity or they can enhance responses induced by pathogen-associated molecules such as LPS. Nucleotides have also been shown to regulate the activity of other transcription factors (AP-1, NFAT, CREB and STAT) in immune and related cell types. Here, we provide an overview of transcription factors shown to be activated by nucleotides in immune cells, and describe what is known about their mechanisms of activation and potential functions. Furthermore, we propose areas for future work in this new and expanding field. PMID- 18404421 TI - The role of P2 receptors in controlling infections by intracellular pathogens. AB - A growing number of studies have demonstrated the importance of ATP(e)-signalling via P2 receptors as an important component of the inflammatory response to infection. More recent studies have shown that ATP(e) can also have a direct effect on infection by intracellular pathogens, by modulating membrane trafficking in cells that contain vacuoles that harbour intracellular pathogens, such as mycobacteria and chlamydiae. A conserved mechanism appears to be involved in controlling infection by both of these pathogens, as a role for phospholipase D in inducing fusion between lysosomes and the vacuoles has been demonstrated. Other P2-dependent mechanisms are most likely operative in the cases of pathogens, such as Leishmania, which survive in an acidic phagolysosomal-like compartment. ATP(e) may function as a "danger signal" that alerts the immune system to the presence of intracellular pathogens that damage the host cell, while different intracellular pathogens have evolved enzymes or other mechanisms to inhibit ATP(e)-mediated signalling, which should, thus, be viewed as virulence factors for these pathogens. PMID- 18404420 TI - Extracellular NAD and ATP: Partners in immune cell modulation. AB - Extracellular NAD and ATP exert multiple, partially overlapping effects on immune cells. Catabolism of both nucleotides by extracellular enzymes keeps extracellular concentrations low under steady-state conditions and generates metabolites that are themselves signal transducers. ATP and its metabolites signal through purinergic P2 and P1 receptors, whereas extracellular NAD exerts its effects by serving as a substrate for ADP-ribosyltransferases (ARTs) and NAD glycohydrolases/ADPR cyclases like CD38 and CD157. Both nucleotides activate the P2X7 purinoceptor, although by different mechanisms and with different characteristics. While ATP activates P2X7 directly as a soluble ligand, activation via NAD occurs by ART-dependent ADP-ribosylation of cell surface proteins, providing an immobilised ligand. P2X7 activation by either route leads to phosphatidylserine exposure, shedding of CD62L, and ultimately to cell death. Activation by ATP requires high micromolar concentrations of nucleotide and is readily reversible, whereas NAD-dependent stimulation begins at low micromolar concentrations and is more stable. Under conditions of cell stress or inflammation, ATP and NAD are released into the extracellular space from intracellular stores by lytic and non-lytic mechanisms, and may serve as "danger signals" to alert the immune response to tissue damage. Since ART expression is limited to naive/resting T cells, P2X7-mediated NAD-induced cell death (NICD) specifically targets this cell population. In inflamed tissue, NICD may inhibit bystander activation of unprimed T cells, reducing the risk of autoimmunity. In draining lymph nodes, NICD may eliminate regulatory T cells or provide space for the preferential expansion of primed cells, and thus help to augment an immune response. PMID- 18404422 TI - The role of released ATP in killing Candida albicans and other extracellular microbial pathogens by cationic peptides. AB - A unifying theme common to the action of many cationic peptides that display lethal activities against microbial pathogens is their specific action at microbial membranes that results in selective loss of ions and small nucleotides chiefly ATP. One model cationic peptide that induces non-lytic release of ATP from the fungal pathogen Candida albicans is salivary histatin 5 (Hst 5). The major characteristic of Hst 5-induced ATP release is that it occurs rapidly while cells are still metabolically active and have polarized membranes, thus precluding cell lysis as the means of release of ATP. Other cationic peptides that induce selective release of ATP from target microbes are lactoferricin, human neutrophil defensins, bactenecin, and cathelicidin peptides. The role of released extracellular ATP induced by cationic peptides is not known, but localized increases in extracellular ATP concentration may serve to potentiate cell killing, facilitate further peptide uptake, or function as an additional signal to activate the host innate immune system at the site of infection. PMID- 18404423 TI - Shaping immune responses through the activation of dendritic cells' P2 receptors. AB - Dendritic cells (DCs) activate and shape the adaptive immune response by capturing antigens, migrating to peripheral lymphoid organs where naive T cells reside, expressing high levels of MHC and costimulatory molecules and secreting cytokines and chemokines. DCs are endowed with a high degree of functional plasticity and their functions are tightly regulated. Besides initiating adaptive immune responses, DCs play a key role in maintaining peripheral tolerance toward self-antigens. On the basis of the information gathered from the tissue where they reside, DCs adjust their functional activity to ensure that protective immunity is favoured while unwanted or exaggerated immune responses are prevented. A wide variety of signals from neighbouring cells affecting DC functional activity have been described. Here we will discuss the complex role of extracellular nucleotides in the regulation of DC function and the role of P2 receptors as possible tools to manipulate immune responses. PMID- 18404424 TI - Adenosine and lymphocyte regulation. AB - Adenosine is a potent extracellular messenger that is produced in high concentrations under metabolically unfavourable conditions. Tissue hypoxia, consequent to a compromised cellular energy status, is followed by the enhanced breakdown of ATP leading to the release of adenosine. Through the interaction with A(2) and A(3) membrane receptors, adenosine is devoted to the restoration of tissue homeostasis, acting as a retaliatory metabolite. Several aspects of the immune response have to be taken into consideration and even though in general it is very important to dampen inflammation, in some circumstances, such as the case of cancer, it is also necessary to increase the activity of immune cells against pathogens. Therefore, adenosine receptors that are defined as "sensors" of metabolic changes in the local tissue environment may be very important targets for modulation of immune responses and drugs devoted to regulating the adenosinergic system are promising in different clinical situations. PMID- 18404426 TI - From "Hellstrom Paradox" to anti-adenosinergic cancer immunotherapy. AB - Cancer therapy by endogenous or adoptively transferred anti-tumor T cells is considered complementary to conventional cancer treatment by surgery, radiotherapy or chemotherapy. However, the scope of promising immunotherapeutic protocols is currently limited because tumors can create a "hostile" immunosuppressive microenvironment that prevents their destruction by anti-tumor T cells. There is a possibility to develop better and more effective immunotherapies by inactivating mechanisms that inhibit anti-tumor T cells in the tumor microenvironment and thereby protect cancerous tissues from immune damage. This may be now possible because of the recent demonstration that genetic deletion of immunosuppressive A2A and A2B adenosine receptors (A2AR and A2BR) or their pharmacological inactivation can prevent the inhibition of anti-tumor T cells by the hypoxic tumor microenvironment and as a result facilitate full tumor rejection [Ohta A, Gorelik E, Prasad SJ et al (2006) Proc Natl Acad Sci USA 103(35):13132-13137]. This approach is based on in vivo genetic evidence that A2AR play a critical role in the protection of normal tissues from overactive immune cells in acutely inflamed and hypoxic areas. The observations of much improved T-cell-mediated rejection of tumors in mice with inactivated A2AR strongly suggest that A2AR also protects hypoxic cancerous tissues and that A2AR should be inactivated in order to improve tumor rejection by anti-tumor T cells. PMID- 18404425 TI - Purinergic modulation of microglial cell activation. AB - Microglial cells are resident macrophages in the brain and their activation is an important part of the brain immune response and the pathology of the major CNS diseases. Microglial activation is triggered by pathological signals and is characterized by morphological changes, proliferation, phagocytosis and the secretion of various cytokines and inflammatory mediators, which could be both destructive and protective for the nervous tissue. Purines are one of the most important mediators which regulate different aspects of microglial function. They could be released to the extracellular space from neurons, astrocytes and from the microglia itself, upon physiological neuronal activity and in response to pathological stimuli and cellular damage. Microglial activation is regulated by various subtypes of nucleotide (P2X, P2Y) and adenosine (A1, A(2A) and A3) receptors, which control ionic conductances, membrane potential, gene transcription, the production of inflammatory mediators and cell survival. Among them, the role of P2X7 receptors is especially well delineated, but P2X4, various P2Y, A1, A(2A) and A3 receptors also powerfully participate in the microglial response. The pathological role of microglial purine receptors has also been demonstrated in disease models; e.g., in ischemia, sclerosis multiplex and neuropathic pain. Due to their upregulation and selective activation under pathological conditions, they provide new avenues in the treatment of neurodegenerative and neuroinflammatory illnesses. PMID- 18404427 TI - P2 receptors and chronic pain. AB - There is abundant evidence that extracellular ATP and other nucleotides have an important role in pain signaling at both the periphery and in the CNS. The focus of attention now is on the possibility that endogenous ATP and its receptor system might be activated in chronic pathological pain states, particularly in neuropathic and inflammatory pain. Neuropathic pain is often a consequence of nerve injury through surgery, bone compression, diabetes or infection. This type of pain can be so severe that even light touching can be intensely painful; unfortunately, this state is generally resistant to currently available treatments. In this review, we summarize the role of ATP receptors, particularly the P2X4, P2X3 and P2X7 receptors, in neuropathic and inflammatory pain. The expression of P2X4 receptors in the spinal cord is enhanced in spinal microglia after peripheral nerve injury, and blocking pharmacologically and suppressing molecularly P2X4 receptors produce a reduction of the neuropathic pain behaviour. Understanding the key roles of these ATP receptors may lead to new strategies for the management of intractable chronic pain. PMID- 18404428 TI - Adenosine in inflammatory joint diseases. AB - Inflammatory joint diseases are a group of heterogeneous disorders with a variety of different etiologies and disease manifestations. However, there are features that are common to all of them: first, the recruitment of various inflammatory cell types that are attracted to involved tissues over the course of the disease process. Second, the treatments used in many of these diseases are commonly medications that suppress or alter immune function. The demonstration that adenosine has endogenous anti-inflammatory functions and that some of the most commonly used anti-rheumatic medications exert their therapeutic effects through stimulation of adenosine release suggest an important role for purinergic signaling in inflammatory rheumatic disorders. PMID- 18404429 TI - P2 receptors in atherosclerosis and postangioplasty restenosis. AB - Atherosclerosis is an immunoinflammatory process that involves complex interactions between the vessel wall and blood components and is thought to be initiated by endothelial dysfunction [Ross (Nature 362:801-809, 1993); Fuster et al. (N Engl J Med 326:242-250, 1992); Davies and Woolf (Br Heart J 69:S3-S11, 1993)]. Extracellular nucleotides that are released from a variety of arterial and blood cells [Di Virgilio and Solini (Br J Pharmacol 135:831-842, 2002)] can bind to P2 receptors and modulate proliferation and migration of smooth muscle cells (SMC), which are known to be involved in intimal hyperplasia that accompanies atherosclerosis and postangioplasty restenosis [Lafont et al. (Circ Res 76:996-1002, 1995)]. In addition, P2 receptors mediate many other functions including platelet aggregation, leukocyte adherence, and arterial vasomotricity. A direct pathological role of P2 receptors is reinforced by recent evidence showing that upregulation and activation of P2Y(2) receptors in rabbit arteries mediates intimal hyperplasia [Seye et al. (Circulation 106:2720-2726, 2002)]. In addition, upregulation of functional P2Y receptors also has been demonstrated in the basilar artery of the rat double-hemorrhage model [Carpenter et al. (Stroke 32:516-522, 2001)] and in coronary artery of diabetic dyslipidemic pigs [Hill et al. (J Vasc Res 38:432-443, 2001)]. It has been proposed that upregulation of P2Y receptors may be a potential diagnostic indicator for the early stages of atherosclerosis [Elmaleh et al. (Proc Natl Acad Sci U S A 95:691-695, 1998)]. Therefore, particular effort must be made to understand the consequences of nucleotide release from cells in the cardiovascular system and the subsequent effects of P2 nucleotide receptor activation in blood vessels, which may reveal novel therapeutic strategies for atherosclerosis and restenosis after angioplasty. PMID- 18404430 TI - The role of P2X7 in pain and inflammation. AB - The P2X 7purinoceptor is unique amongst the P2X receptor family in that its activation is able to stimulate the release of mature, biologically active interleukin-1beta (IL-1beta), as well as a variety of other proinflammatory cytokines. Coupled with the predominate localisation of this receptor to immunocytes of haemopoetic origin, this receptor is an obvious candidate to play a major and pivotal role in processes of pain and inflammation. Using genetically modified animals that lack the P2X7 receptor, several investigators have shown that these mice do indeed demonstrate a blunted inflammatory response, and fail to develop pain following both inflammatory and neuropathic insult. These animals also show altered cytokine production in response to inflammatory stimulus, which is far broader than merely modulation of IL-1beta release. In this short article, we review the role of the P2X7 receptor in modulating the release of cytokines and other mediators, and discuss the findings made from P2X7 receptor-deficient animals. As well as highlighting outstanding questions regarding this intriguing receptor, we also speculate as to the potential therapeutic benefit of P2X7 receptor modulation. PMID- 18404431 TI - CD39 and control of cellular immune responses. AB - CD39 is the cell surface-located prototypic member of the ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) family. Biological actions of CD39 are a consequence (at least in part) of the regulated phosphohydrolytic activity on extracellular nucleotides. This ecto-enzymatic cascade in tandem with CD73 (ecto-5'-nucleotidase) also generates adenosine and has major effects on both P2 and adenosine receptor signalling. Despite the early recognition of CD39 as a B lymphocyte activation marker, little is known of the role of CD39 in humoral or cellular immune responses. There is preliminary evidence to suggest that CD39 may impact upon antibody affinity maturation. Pericellular nucleotide/nucleoside fluxes caused by dendritic cell expressed CD39 are also involved in the recruitment, activation and polarization of naive T cells. We have recently explored the patterns of CD39 expression and the functional role of this ecto nucleotidase within quiescent and activated T cell subsets. Our data indicate that CD39, together with CD73, efficiently distinguishes T regulatory cells (Treg) from other resting or activated T cells in mice (and humans). Furthermore, CD39 serves as an integral component of the suppressive machinery of Treg, acting, at least in part, through the modulation of pericellular levels of adenosine. We have also shown that the coordinated regulation of CD39/CD73 expression and of the adenosine receptor A2A activates an immunoinhibitory loop that differentially regulates Th1 and Th2 responses. The in vivo relevance of this network is manifest in the phenotype of Cd39-null mice that spontaneously develop features of autoimmune diseases associated with Th1 immune deviation. These data indicate the potential of CD39 and modulated purinergic signalling in the co-ordination of immunoregulatory functions of dendritic and Treg cells. Our findings also suggest novel therapeutic strategies for immune-mediated diseases. PMID- 18404432 TI - Pyrazolo-triazolo-pyrimidines as adenosine receptor antagonists: A complete structure-activity profile. AB - In the last 5 years, many efforts have been conducted searching potent and selective human A(3) adenosine antagonists. In this field several different classes of compounds, possessing very good affinity (nM range) and with a broad range of selectivity, have been proposed. Recently, our group synthesized a new series of pyrazolo-triazolo-pyrimidines bearing different substitutions at the N(5) and N(8) positions, which have been described as highly potent and selective human A(3) adenosine receptor antagonists. The present review summarizes available data and provides an overview of the structure-activity relationships found for this class of human A(3) adenosine receptor antagonists. PMID- 18404433 TI - Residual platelet ADP reactivity after clopidogrel treatment is dependent on activation of both the unblocked P2Y(1) and the P2Y (12) receptor and is correlated with protein expression of P2Y (12). AB - Two ADP receptors have been identified on human platelets: P2Y(1) and P2Y(12). The P2Y(12) receptor blocker clopidogrel is widely used to reduce the risks in acute coronary syndromes, but, currently, there is no P2Y(1) blocker in clinical use. Evidence for variable responses to clopidogrel has been described in several reports. The mechanistic explanation for this phenomenon is not fully understood. The aim of this study was to examine mechanisms responsible for variability of 2MeS-ADP, a stable ADP analogue, induced platelet reactivity in clopidogrel treated patients. Platelet reactivity was assessed by flow cytometry measurements of P-selectin (CD62P) and activated GpIIb/IIIa complex (PAC-1). Residual 2MeS-ADP activation via the P2Y(12) and P2Y(1) receptors was determined by co-incubation with the selective antagonists AR-C69931 and MRS2179 in vitro. P2Y(1) and P2Y(12) receptor expression on both RNA and protein level were determined, as well as the P2Y(12) H1 or H2 haplotypes. Our data suggest that the residual platelet activation of 2MeS-ADP after clopidogrel treatment is partly due to an inadequate antagonistic effect of clopidogrel on the P2Y(12) receptor and partly due to activation of the P2Y(1) receptor, which is unaffected by clopidogrel. Moreover, a correlation between increased P2Y(12) protein expression on platelets and decreased response to clopidogrel was noticed, r(2)=0.43 (P<0.05). No correlation was found between P2Y(12) mRNA levels and clopidogrel resistance, indicating post transcriptional mechanisms. To achieve additional ADP inhibition in platelets, antagonists directed at the P2Y(1) receptor could be more promising than the development of more potent P2Y(12) receptor antagonists. PMID- 18404435 TI - P2Y(1) and P2Y(12) receptor cross-talk in calcium signalling: Evidence from nonstarved and long-term serum-deprived glioma C6 cells. AB - The current work presents results of experiments on the calcium response evoked by the stimulation by extracellular nucleotides occurring in control, nonstarved glioma C6 cells and in cells after long-term (96 h) serum starvation. Three nucleotide receptors were studied: P2Y(1), P2Y(2) and P2Y(12). Two of them, P2Y(1) and P2Y(2), directly stimulate calcium response. The protein level of the P2Y(2) receptor did not change during the serum starvation, while P2Y(1) protein level fell dramatically. Observed changes in the calcium response generated by P2Y(1) are directly correlated with the receptor protein level as well as with the amount of calcium present in the intracellular calcium stores, partially depleted during starvation process. The third receptor, P2Y(12), did not directly evoke calcium response, however it is activated by the same ligand as P2Y(1). The experiments with AR-C69941MX, the P2Y(12)-specific antagonist, indicated that in control and serum-starved cells, calcium response evoked by P2Y(1) receptor is potentiated by the activity of P2Y(12)-dependent signaling pathways. This potentiation may be mediated by P2Y(12) inhibitory effect on the plasma membrane calcium pump. The calcium influx enhanced by the cooperation of P2Y(1) and P2Y(12) receptor activity directly depends on the capacitative calcium entrance mechanism. PMID- 18404434 TI - Role of P2 purinergic receptors in synaptic transmission under normoxic and ischaemic conditions in the CA1 region of rat hippocampal slices. AB - The role of ATP and its stable analogue ATPgammaS [adenosine-5'-o-(3 thio)triphosphate] was studied in rat hippocampal neurotransmission under normoxic conditions and during oxygen and glucose deprivation (OGD). Field excitatory postsynaptic potentials (fEPSPs) from the dendritic layer or population spikes (PSs) from the soma were extracellularly recorded in the CA1 area of the rat hippocampus. Exogenous application of ATP or ATPgammaS reduced fEPSP and PS amplitudes. In both cases the inhibitory effect was blocked by the selective A(1) adenosine receptor antagonist DPCPX (8-cyclopentyl-1,3 dipropylxanthine) and was potentiated by different ecto-ATPase inhibitors: ARL 67156 (6-N,N-diethyl-D: -beta,gamma-dibromomethylene), BGO 136 (1 hydroxynaphthalene-3,6-disulfonate) and PV4 [hexapotassium dihydrogen monotitanoundecatungstocobaltate(II) tridecahydrate, K(6)H(2)[TiW(11)CoO(40)].13H(2)O]. ATPgammaS-mediated inhibition was reduced by the P2 antagonist suramin [8-(3-benzamido-4-methylbenzamido)naphthalene-1,3,5 trisulfonate] at the somatic level and by other P2 blockers, PPADS (pyridoxalphosphate-6-azophenyl-2',4'-disulfonate) and MRS 2179 (2'-deoxy-N (6) methyladenosine 3',5'-bisphosphate), at the dendritic level. After removal of both P2 agonists, a persistent increase in evoked synaptic responses was recorded both at the dendritic and somatic levels. This effect was prevented in the presence of different P2 antagonists. A 7-min OGD induced tissue anoxic depolarization and was invariably followed by irreversible loss of fEPSP. PPADS, suramin, MRS2179 or BBG (brilliant blue G) significantly prevented the irreversible failure of neurotransmission induced by 7-min OGD. Furthermore, in the presence of these P2 antagonists, the development of anoxic depolarization was blocked or significantly delayed. Our results indicate that P2 receptors modulate CA1 synaptic transmission under normoxic conditions by eliciting both inhibitory and excitatory effects. In the same brain region, P2 receptor stimulation plays a deleterious role during a severe OGD insult. PMID- 18404436 TI - On the history of ecto-ATPases: The role of W. A. Engelhardt. PMID- 18404438 TI - Second Joint Italian-German Purine Club Meeting: "Progress in purinergic receptor pharmacology and function". PMID- 18404437 TI - Ecto-5'-nucleotidase and intestinal ion secretion by enteropathogenic Escherichia coli. AB - Enteropathogenic Escherichia coli (EPEC) triggers a large release of adenosine triphosphate (ATP) from host intestinal cells and the extracellular ATP is broken down to adenosine diphosphate (ADP), AMP, and adenosine. Adenosine is a potent secretagogue in the small and large intestine. We suspected that ecto-5' nucleotidase (CD73, an intestinal enzyme) was a critical enzyme involved in the conversion of AMP to adenosine and in the pathogenesis of EPEC diarrhea. We developed a nonradioactive method for measuring ecto-5'-nucleotidase in cultured T84 cell monolayers based on the detection of phosphate release from 5'-AMP. EPEC infection triggered a release of ecto-5'-nucleotidase from the cell surface into the supernatant medium. EPEC-induced 5'-nucleotidase release was not correlated with host cell death but instead with activation of phosphatidylinositol-specific phospholipase C (PI-PLC). Ecto-5'-nucleotidase was susceptible to inhibition by zinc acetate and by alpha,beta-methylene-adenosine diphosphate (alpha,beta methylene-ADP). In the Ussing chamber, these inhibitors could reverse the chloride secretory responses triggered by 5'-AMP. In addition, alpha,beta methylene-ADP and zinc blocked the ability of 5'-AMP to stimulate EPEC growth under nutrient-limited conditions in vitro. Ecto-5'-nucleotidase appears to be the major enzyme responsible for generation of adenosine from adenine nucleotides in the T84 cell line, and inhibitors of ecto-5'-nucleotidase, such as alpha,beta methylene-ADP and zinc, might be useful for treatment of the watery diarrhea produced by EPEC infection. PMID- 18404439 TI - Activation kinetics of single P2X receptors. AB - After the primary structure of P2X receptors had been identified, their function had to be characterized on the molecular level. Since these ligand-gated ion channels become activated very quickly after binding of ATP, methods with adequate time resolution have to be applied to investigate the early events induced by the agonist. Single-channel recordings were performed to describe conformational changes on P2X(2), P2X(4), and P2X(7) receptors induced by ATP and also by allosteric receptor modifiers. The main results of these studies and the models of P2X receptor kinetics derived from these observations are reviewed here. The investigation of purinoceptors by means of the patch clamp technique following site-directed mutagenesis will probably reveal more details of P2X receptor function at the molecular level. PMID- 18404440 TI - Structural and functional evolution of the P2Y(12)-like receptor group. AB - Metabotropic pyrimidine and purine nucleotide receptors (P2Y receptors) belong to the superfamily of G protein-coupled receptors (GPCR). They are distinguishable from adenosine receptors (P1) as they bind adenine and/or uracil nucleotide triphosphates or diphosphates depending on the subtype. Over the past decade, P2Y receptors have been cloned from a variety of tissues and species, and as many as eight functional subtypes have been characterized. Most recently, several members of the P2Y(12)-like receptor group, which includes the clopidogrel-sensitive ADP receptor P2Y(12), have been deorphanized. The P2Y(12)-like receptor group comprises several structurally related GPCR which, however, display heterogeneous agonist specificity including nucleotides, their derivatives, and lipids. Besides the established function of P2Y(12) in platelet activation, expression in macrophages, neuronal and glial cells as well as recent results from functional studies implicate that several members of this group may have specific functions in neurotransmission, inflammation, chemotaxis, and response to tissue injury. This review focuses specifically on the structure-function relation and shortly summarizes some aspects of the physiological relevance of P2Y(12)-like receptor members. PMID- 18404441 TI - P2 receptor-mediated modulation of neurotransmitter release-an update. AB - Presynaptic nerve terminals are equipped with a number of presynaptic auto- and heteroreceptors, including ionotropic P2X and metabotropic P2Y receptors. P2 receptors serve as modulation sites of transmitter release by ATP and other nucleotides released by neuronal activity and pathological signals. A wide variety of P2X and P2Y receptors expressed at pre- and postsynaptic sites as well as in glial cells are involved directly or indirectly in the modulation of neurotransmitter release. Nucleotides are released from synaptic and nonsynaptic sites throughout the nervous system and might reach concentrations high enough to activate these receptors. By providing a fine-tuning mechanism these receptors also offer attractive sites for pharmacotherapy in nervous system diseases. Here we review the rapidly emerging data on the modulation of transmitter release by facilitatory and inhibitory P2 receptors and the receptor subtypes involved in these interactions. PMID- 18404442 TI - Adenosine A(1) receptor: Functional receptor-receptor interactions in the brain. AB - Over the past decade, many lines of investigation have shown that receptor mediated signaling exhibits greater diversity than previously appreciated. Signal diversity arises from numerous factors, which include the formation of receptor dimers and interplay between different receptors. Using adenosine A(1) receptors as a paradigm of G protein-coupled receptors, this review focuses on how receptor receptor interactions may contribute to regulation of the synaptic transmission within the central nervous system. The interactions with metabotropic dopamine, adenosine A(2A), A(3), neuropeptide Y, and purinergic P2Y(1) receptors will be described in the first part. The second part deals with interactions between A(1)Rs and ionotropic receptors, especially GABA(A), NMDA, and P2X receptors as well as ATP-sensitive K(+) channels. Finally, the review will discuss new approaches towards treating neurological disorders. PMID- 18404443 TI - The role of ATP and adenosine in the brain under normoxic and ischemic conditions. AB - By taking advantage of some recently synthesized compounds that are able to block ecto-ATPase activity, we demonstrated that adenosine triphosphate (ATP) in the hippocampus exerts an inhibitory action independent of its degradation to adenosine. In addition, tonic activation of P2 receptors contributes to the normally recorded excitatory neurotransmission. The role of P2 receptors becomes critical during ischemia when extracellular ATP concentrations increase. Under such conditions, P2 antagonism is protective. Although ATP exerts a detrimental role under ischemia, it also exerts a trophic role in terms of cell division and differentiation. We recently reported that ATP is spontaneously released from human mesenchymal stem cells (hMSCs) in culture. Moreover, it decreases hMSC proliferation rate at early stages of culture. Increased hMSC differentiation could account for an ATP-induced decrease in cell proliferation. ATP as a homeostatic regulator might exert a different effect on cell trophism according to the rate of its efflux and receptor expression during the cell life cycle. During ischemia, adenosine formed by intracellular ATP escapes from cells through the equilibrative transporter. The protective role of adenosine A(1) receptors during ischemia is well accepted. However, the use of selective A(1) agonists is hampered by unwanted peripheral effects, thus attention has been focused on A(2A) and A(3) receptors. The protective effects of A(2A) antagonists in brain ischemia may be largely due to reduced glutamate outflow from neurones and glial cells. Reduced activation of p38 mitogen-activated protein kinases that are involved in neuronal death through transcriptional mechanisms may also contribute to protection by A(2A) antagonism. Evidence that A(3) receptor antagonism may be protective after ischemia is also reported. PMID- 18404444 TI - Modification of neuropathic pain sensation through microglial ATP receptors. AB - Neuropathic pain that typically develops when peripheral nerves are damaged through surgery, bone compression in cancer, diabetes, or infection is a major factor causing impaired quality of life in millions of people worldwide. Recently, there has been a rapidly growing body of evidence indicating that spinal glia play a critical role in the pathogenesis of neuropathic pain. Accumulating findings also indicate that nucleotides play an important role in neuron-glia communication through P2 purinoceptors. Damaged neurons release or leak nucleotides including ATP and UTP to stimulate microglia through P2 purinoceptors expressing on microglia. It was shown in an animal model of neuropathic pain that microglial P2X(4) and P2X(7) receptors are crucial in pain signaling after peripheral nerve lesion. In this review, we describe the modification of neuropathic pain sensation through microglial P2X(4) and P2X(7), with the possibility of P2Y(6) and P2Y(12) involvement. PMID- 18404446 TI - Adenosine A(2A) receptors modulate BDNF both in normal conditions and in experimental models of Huntington's disease. AB - Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, enhances synaptic transmission and regulates neuronal proliferation and survival. Functional interactions between adenosine A(2A) receptors (A(2A)Rs) and BDNF have been recently reported. In this article, we report some recent findings from our group showing that A(2A)Rs regulate both BDNF functions and levels in the brain. Whereas BDNF (10 ng/ml) increased the slope of excitatory postsynaptic field potentials (fEPSPs) in hippocampal slices from wild-type (WT) mice, it was completely ineffective in slices taken from A(2A)R knock-out (KO) mice. Furthermore, enzyme immunoassay studies showed a significant reduction in hippocampal BDNF levels in A(2A)R KO vs. WT mice. Having found an even marked reduction in the striatum of A(2A)R KO mice, and as both BDNF and A(2A)Rs have been implicated in the pathogenesis of Huntington's disease (HD), an inherited striatal neurodegenerative disease, we then evaluated whether the pharmacological blockade of A(2A)Rs could influence striatal levels of BDNF in an experimental model of HD-like striatal degeneration (quinolinic acid-lesioned rats) and in a transgenic mice model of HD (R6/2 mice). In both QA-lesioned rats and early symptomatic R6/2 mice (8 weeks), the systemic administration of the A(2A)R antagonist SCH58261 significantly reduced striatal BDNF levels. These results indicate that the presence and the tonic activation of A(2A)Rs are necessary to allow BDNF-induced potentiation of synaptic transmission and to sustain a normal BDNF tone. The possible functional consequences of reducing striatal BDNF levels in HD models need further investigation. PMID- 18404445 TI - New insights into purinergic receptor signaling in neuronal differentiation, neuroprotection, and brain disorders. AB - Ionotropic P2X and metabotropic P2Y purinergic receptors are expressed in the central nervous system and participate in the synaptic process particularly associated with acetylcholine, GABA, and glutamate neurotransmission. As a result of activation, the P2 receptors promote the elevation of free intracellular calcium concentration as the main signaling pathway. Purinergic signaling is present in early stages of embryogenesis and is involved in processes of cell proliferation, migration, and differentiation. The use of new techniques such as knockout animals, in vitro models of neuronal differentiation, antisense oligonucleotides to induce downregulation of purinergic receptor gene expression, and the development of selective inhibitors for purinergic receptor subtypes contribute to the comprehension of the role of purinergic signaling during neurogenesis. In this review, we shall discuss the participation of purinergic receptors in developmental processes and in brain physiology, including neuron glia interactions and pathophysiology. PMID- 18404447 TI - New 2,6,9-trisubstituted adenines as adenosine receptor antagonists: a preliminary SAR profile. AB - A new series of 2,6,9-trisubstituted adenines (5-14) have been prepared and evaluated in radioligand binding studies for their affinity at the human A(1), A(2A) and A(3) adenosine receptors and in adenylyl cyclase experiments for their potency at the human A(2B) subtype. From this preliminary study the conclusion can be drawn that introduction of bulky chains at the N (6) position of 9 propyladenine significantly increased binding affinity at the human A(1) and A(3) adenosine receptors, while the presence of a chlorine atom at the 2 position resulted in a not univocal effect, depending on the receptor subtype and/or on the substituent present in the N (6) position. However, in all cases, the presence in the 2 position of a chlorine atom favoured the interaction with the A(2A) subtype. These results demonstrated that, although the synthesized compounds were found to be quite inactive at the human A(2B) subtype, adenine is a useful template for further development of simplified adenosine receptor antagonists with distinct receptor selectivity profiles. PMID- 18404448 TI - [(3)H]Adenine is a suitable radioligand for the labeling of G protein-coupled adenine receptors but shows high affinity to bacterial contaminations in buffer solutions. AB - [(3)H]Adenine has previously been used to label the newly discovered G protein coupled murine adenine receptors. Recent reports have questioned the suitability of [(3)H]adenine for adenine receptor binding studies because of curious results, e.g. high specific binding even in the absence of mammalian protein. In this study, we showed that specific [(3)H]adenine binding to various mammalian membrane preparations increased linearly with protein concentration. Furthermore, we found that Tris-buffer solutions typically used for radioligand binding studies (50 mM, pH 7.4) that have not been freshly prepared but stored at 4 degrees C for some time may contain bacterial contaminations that exhibit high affinity binding for [(3)H]adenine. Specific binding is abolished by heating the contaminated buffer or filtering it through 0.2-mum filters. Three different, aerobic, gram-negative bacteria were isolated from a contaminated buffer solution and identified as Achromobacter xylosoxidans, A. denitrificans, and Acinetobacter lwoffii. A. xylosoxidans, a common bacterium that can cause nosocomial infections, showed a particularly high affinity for [(3)H]adenine in the low nanomolar range. Structure-activity relationships revealed that hypoxanthine also bound with high affinity to A. xylosoxidans, whereas other nucleobases (uracil, xanthine) and nucleosides (adenosine, uridine) did not. The nature of the labeled site in bacteria is not known, but preliminary results indicate that it may be a high-affinity purine transporter. We conclude that [(3)H]adenine is a well suitable radioligand for adenine receptor binding studies but that bacterial contamination of the employed buffer solutions must be avoided. PMID- 18404449 TI - Monitoring the expression of purinoceptors and nucleotide-metabolizing ecto enzymes with antibodies directed against proteins in native conformation. AB - Following their release from cells, ATP and NAD, the universal currencies of energy metabolism, function as extracellular signalling molecules. Mammalian cells express numerous purinoceptors, i.e., the nucleotide-gated P2X ion channels and the G-protein-coupled P2Y receptors. Signalling through purinoceptors is controlled by nucleotide-metabolizing ecto-enzymes, which regulate the availability of extracellular nucleotides. These enzymes include ecto-nucleoside triphosphate diphosphohydrolases (ENTPD, CD39 family) and ecto-nucleotide pyrophosphatase/phosphodiesterases (ENPP, CD203 family). Investigation of these receptors and enzymes has been hampered by the lack of available antibodies, especially ones that recognize these proteins in their native conformation. This study reports the use of genetic immunization to generate such antibodies against P2X(1), P2X(4), P2X(7), ENTPD1, ENPTD2, ENPTD5, ENPTD6, ENPP2, ENPP3, ENPP4, ENPP5, and ENPP6. Genetic immunization ensures expression of the native protein by the cells of the immunized animal and yields antibodies directed against proteins in native conformation (ADAPINCs). Such antibodies are especially useful for immunofluorescence and immunoprecipitation analyses, whereas antibodies against synthetic peptides usually function well only in Western-blot analyses. Here we illustrate the utility of the new antibodies to monitor the cell surface expression of and to purify some key players of purinergic signalling. PMID- 18404450 TI - Autoregulation in PC12 cells via P2Y receptors: Evidence for non-exocytotic nucleotide release from neuroendocrine cells. AB - Nucleotides are released not only from neurons, but also from various other types of cells including fibroblasts, epithelial, endothelial and glial cells. While ATP release from non-neural cells is frequently Ca(2+) independent and mostly non vesicular, neuronal ATP release is generally believed to occur via exocytosis. To evaluate whether nucleotide release from neuroendocrine cells might involve a non vesicular component, the autocrine/paracrine activation of P2Y(12) receptors was used as a biosensor for nucleotide release from PC12 cells. Expression of a plasmid coding for the botulinum toxin C1 light chain led to a decrease in syntaxin 1 detected in immunoblots of PC12 membranes. In parallel, spontaneous as well as depolarization-evoked release of previously incorporated [(3)H]noradrenaline from transfected cells was significantly reduced in comparison with the release from untransfected cells, thus indicating that exocytosis was impaired. In PC12 cells expressing the botulinum toxin C1 light chain, ADP reduced cyclic AMP synthesis to the same extent as in non-transfected cells. Likewise, the enhancement of cyclic AMP synthesis either due to the blockade of P2Y(12) receptors or due to the degradation of extracellular neucleotides by apyrase was not different between non-transfected and botulinum toxin C1 light chain expressing cells. However, the inhibition of cyclic AMP synthesis caused by depolarization-evoked release of endogenous nucleotides was either abolished or greatly reduced in cells expressing the botulinum toxin C1 light chain. Together, these results show that spontaneous nucleotide release from neuroendocrine cells may occur independently of vesicle exocytosis, whereas depolarization-evoked nucleotide release relies predominantly on exocytotic mechanisms. PMID- 18404451 TI - Lack of evidence for direct phosphorylation of recombinantly expressed P2X(2) and P2X (3) receptors by protein kinase C. AB - P2X(3) and P2X(2+3) receptors are present on sensory neurons, where they contribute not only to transient nociceptive responses, but also to hypersensitivity underlying pathological pain states elicited by nerve injuries. Increased signalling through P2X(3) and P2X(2+3) receptors may arise from an increased routing to the plasma membrane and/or gain of function of pre-existing receptors. An obvious effector mechanism for functional modulation is protein kinase C (PKC)-mediated phosphorylation, since all P2X family members share a conserved consensus sequence for PKC, TXR/K, within the intracellularly located N terminal domain. Contradictory reports have been published regarding the exact role of this motif. In the present study, we confirm that site-directed elimination of the potential phosphor-acceptor threonine or the basic residue in the P+2 position of the TXR/K sequence accelerates desensitization of P2X(2) receptors and abolishes P2X(3) receptor function. Moreover, the PKC activator phorbol 12-myristate 13-acetate increased P2X(3) (but not P2X(2)) receptor mediated currents. Biochemically, however, we were unable to demonstrate by various experimental approaches a direct phosphorylation of wild-type P2X(2) and P2X(3) receptors expressed in both Xenopus laevis oocytes and HEK293 cells. In conclusion, our data support the view that the TXR/K motif plays an important role in P2X function and that phorbol 12-myristate 13-acetate is capable of modulating some P2X receptor subtypes. The underlying mechanism, however, is unlikely to involve direct PKC-mediated P2X receptor phosphorylation. PMID- 18404452 TI - Mapping P2X and P2Y receptor proteins in striatum and substantia nigra: An immunohistological study. AB - Our work aimed to provide a topographical analysis of all known ionotropic P2X(1 7) and metabotropic P2Y(1,2,4,6,11-14) receptors that are present in vivo at the protein level in the basal ganglia nuclei and particularly in rat brain slices from striatum and substantia nigra. By immunohistochemistry-confocal and Western blotting techniques, we show that, with the exception of P2Y(11,13) receptors, all other subtypes are specifically expressed in these areas in different amounts, with ratings of low (P2X(5,6) and P2Y(1,6,14) in striatum), medium (P2X(3) in striatum and substantia nigra, P2X(6,7) and P2Y(1) in substantia nigra) and high. Moreover, we describe that P2 receptors are localized on neurons (colocalizing with neurofilament light, medium and heavy chains) with features that are either dopaminergic (colocalizing with tyrosine hydroxylase) or GABAergic (colocalizing with parvalbumin and calbindin), and they are also present on astrocytes (P2Y(2,4), colocalizing with glial fibrillary acidic protein). In addition, we aimed to investigate the expression of P2 receptors after dopamine denervation, obtained by using unilateral injection of 6 hydroxydopamine as an animal model of Parkinson's disease. This generates a rearrangement of P2 proteins: most P2X and P2Y receptors are decreased on GABAergic and dopaminergic neurons, in the lesioned striatum and substantia nigra, respectively, as a consequence of dopaminergic denervation and/or neuronal degeneration. Conversely, P2X(1,3,4,6) on GABAergic neurons and P2Y(4) on astrocytes augment their expression exclusively in the lesioned substantia nigra reticulata, probably as a compensatory reaction to dopamine shortage. These results disclose the presence of P2 receptors in the normal and lesioned nigro striatal circuit, and suggest their potential participation in the mechanisms of Parkinson's disease. PMID- 18404453 TI - MPP(+)-induced cytotoxicity in neuroblastoma cells: Antagonism and reversal by guanosine. AB - Guanosine exerts neuroprotective effects in the central nervous system. Apoptosis, a morphological form of programmed cell death, is implicated in the pathophysiology of Parkinson's disease (PD). MPP(+), a dopaminergic neurotoxin, produces in vivo and in vitro cellular changes characteristic of PD, such as cytotoxicity, resulting in apoptosis. Undifferentiated human SH-SY5Y neuroblastoma cells had been used as an in vitro model of Parkinson's disease. We investigated if extracellular guanosine affected MPP(+)-induced cytotoxicity and examined the molecular mechanisms mediating its effects. Exposure of neuroblastoma cells to MPP(+) (10 muM-5 mM for 24-72 h) induced DNA fragmentation in a time-dependent manner (p < 0.05). Administration of guanosine (100 muM) before, concomitantly with or, importantly, after the addition of MPP(+) abolished MPP(+)-induced DNA fragmentation. Addition of MPP(+) (500 muM) to cells increased caspase-3 activity over 72 h (p < 0.05), and this was abolished by pre- or co-treatment with guanosine. Exposure of cells to pertussis toxin prior to MPP(+) eliminated the anti-apoptotic effect of guanosine, indicating that this effect is dependent on a Gi protein-coupled receptor, most likely the putative guanosine receptor. The protection by guanosine was also abolished by the selective inhibitor of the enzyme PI-3-K/Akt/PKB (LY294002), confirming that this pathway plays a decisive role in this effect of guanosine. Neither MPP(+) nor guanosine had any significant effect on alpha-synuclein expression. Thus, guanosine antagonizes and reverses MPP(+)-induced cytotoxicity of neuroblastoma cells via activation of the cell survival pathway, PI-3-K/Akt/PKB. Our results suggest that guanosine may be an effective pharmacological intervention in PD. PMID- 18404454 TI - Guanosine reduces apoptosis and inflammation associated with restoration of function in rats with acute spinal cord injury. AB - Spinal cord injury results in progressive waves of secondary injuries, cascades of noxious pathological mechanisms that substantially exacerbate the primary injury and the resultant permanent functional deficits. Secondary injuries are associated with inflammation, excessive cytokine release, and cell apoptosis. The purine nucleoside guanosine has significant trophic effects and is neuroprotective, antiapoptotic in vitro, and stimulates nerve regeneration. Therefore, we determined whether systemic administration of guanosine could protect rats from some of the secondary effects of spinal cord injury, thereby reducing neurological deficits. Systemic administration of guanosine (8 mg/kg per day, i.p.) for 14 consecutive days, starting 4 h after moderate spinal cord injury in rats, significantly improved not only motor and sensory functions, but also recovery of bladder function. These improvements were associated with reduction in the inflammatory response to injury, reduction of apoptotic cell death, increased sparing of axons, and preservation of myelin. Our data indicate that the therapeutic action of guanosine probably results from reducing inflammation resulting in the protection of axons, oligodendrocytes, and neurons and from inhibiting apoptotic cell death. These data raise the intriguing possibility that guanosine may also be able to reduce secondary pathological events and thus improve functional outcome after traumatic spinal cord injury in humans. PMID- 18404455 TI - Ectonucleotidases in Muller glial cells of the rodent retina: Involvement in inhibition of osmotic cell swelling. AB - Extracellular nucleotides mediate glia-to-neuron signalling in the retina and are implicated in the volume regulation of retinal glial (Muller) cells under osmotic stress conditions. We investigated the expression and functional role of ectonucleotidases in Muller cells of the rodent retina by cell-swelling experiments, calcium imaging, and immuno- and enzyme histochemistry. The swelling of Muller cells under hypoosmotic stress was inhibited by activation of an autocrine purinergic signalling cascade. This cascade is initiated by exogenous glutamate and involves the consecutive activation of P2Y(1) and adenosine A1 receptors, the action of ectoadenosine 5'-triphosphate (ATP)ases, and a nucleoside-transporter-mediated release of adenosine. Inhibition of ectoapyrases increased the ATP-evoked calcium responses in Muller cell endfeet. Muller cells were immunoreactive for nucleoside triphosphate diphosphohydrolases (NTPDase)2 (but not NTPDase1), ecto-5'-nucleotidase, P2Y(1), and A1 receptors. Enzyme histochemistry revealed that ATP but not adenosine 5'-diphosphate (ADP) is extracellularly metabolised in retinal slices of NTPDase1 knockout mice. NTPDase1 activity and protein is restricted to blood vessels, whereas activity of alkaline phosphatase is essentially absent at physiological pH. The data suggest that NTPDase2 is the major ATP-degrading ectonucleotidase of the retinal parenchyma. NTPDase2 expressed by Muller cells can be implicated in the regulation of purinergic calcium responses and cellular volume. PMID- 18404456 TI - Involvement of P2X and P2Y receptors in microglial activation in vivo. AB - Microglial cells are the primary immune effector cells in the brain. Extracellular ATP, e.g., released after brain injury, may initiate microglial activation via stimulation of purinergic receptors. In the rat nucleus accumbens (NAc), the involvement of P2X and P2Y receptors in the generation of microglial reaction in vivo was investigated. A stab wound in the NAc increased immunoreactivity (IR) for P2X(1,2,4,7) and P2Y(1,2,4,6,12) receptors on microglial cells when visualized with confocal laser scanning microscopy. A prominent immunolabeling of P2X(7) receptors with antibodies directed against the ecto- or endodomain was found on Griffonia simplicifolia isolectin-B4-positive cells. Additionally, the P2X(7) receptor was colocalized with active caspase 3 but not with the anti-apoptotic marker pAkt. Four days after local application of the agonists alpha,betameATP, ADPbetaS, 2MeSATP, and BzATP, an increase in OX 42- and G. simplicifolia isolectin-IR was observed around the stab wound, quantified both densitometrically and by counting the number of ramified and activated microglial cells, whereas UTPgammaS appeared to be ineffective. The P2 receptor antagonists PPADS and BBG decreased the injury-induced increase of these IRs when given alone and in addition inhibited the agonist effects. Further, the intra accumbally applied P2X(7) receptor agonist BzATP induced an increase in the number of caspase-3-positive cells. These results indicate that ATP, acting via different P2X and P2Y receptors, is a signaling molecule in microglial cell activation after injury in vivo. The up-regulation of P2X(7)-IR after injury suggests that this receptor is involved in apoptotic rather than proliferative effects. PMID- 18404457 TI - Role of A(2A) adenosine receptors in regulation of opsonized E. coli-induced macrophage function. AB - Adenosine is a biologically active molecule that is formed at sites of metabolic stress associated with trauma and inflammation, and its systemic level reaches high concentrations in sepsis. We have recently shown that inactivation of A(2A) adenosine receptors decreases bacterial burden as well as IL-10, IL-6, and MIP-2 production in mice that were made septic by cecal ligation and puncture (CLP). Macrophages are important in both elimination of pathogens and cytokine production in sepsis. Therefore, in the present study, we questioned whether macrophages are responsible for the decreased bacterial load and cytokine production in A(2A) receptor-inactivated septic mice. We showed that A(2A) KO and WT peritoneal macrophages obtained from septic animals were equally effective in phagocytosing opsonized E. coli. IL-10 production induced by opsonized E. coli was decreased in macrophages obtained from septic A(2A) KO mice as compared to WT counterparts. In contrast, the release of IL-6 and MIP-2 induced by opsonized E. coli was higher in septic A(2A) KO macrophages than WT macrophages. These results suggest that peritoneal macrophages are not responsible for the decreased bacterial load and diminished MIP-2 and IL-6 production that are observed in septic A(2A) KO mice. In contrast, peritoneal macrophages may contribute to the suppressive effect of A(2A) receptor inactivation on IL-10 production during sepsis. PMID- 18404458 TI - First joint italian-german purine club meeting "progress in purinergic receptor pharmacology and function". PMID- 18404459 TI - The P2Y4 receptor forms homo-oligomeric complexes in several CNS and PNS neuronal cells. AB - It is well established that several cell surface receptors interact with each other to form dimers and oligomers, which are essential for their activation. Since little is known about the quaternary structure of P2Y receptors, in the present work, we investigated the expression of the G-protein-coupled P2Y4 subunit as monomeric or higher-order complex protein. We examined both endogenously expressed P2Y4 subtype with the aid of specific anti-P2Y4 antiserum, and heterologously transfected P2Y4-tagged receptors with the use of antitag antibodies. In both cases, we found the P2Y4 receptor displaying molecular masses corresponding to monomeric, dimeric and oligomeric structures. Experiments performed in the absence of reducing agents demonstrated that there is a strict correlation among the multiple protein bands and that the multimeric forms are at least partially assembled by disulphide bonds. The direct demonstration of P2Y4 homodimerisation comes instead from co-transfection and differential co immunoprecipitation experiments, with the use of differently tagged P2Y4 receptors and antitag antibodies. The structural propensity of the P2Y4 protein to form homo-oligomers may open the possibility of a novel regulatory mechanism of physiopathological functions for this and additional P2Y receptors. PMID- 18404460 TI - Novel selective antagonist radioligands for the pharmacological study of A(2B) adenosine receptors. AB - The adenosine A(2B) receptor is the least well characterized of the four adenosine subtypes due to the lack of potent and selective agonists and antagonists. Despite the widespread distribution of A(2B) receptor mRNA, little information is available with regard to their function. The characterization of A(2B) receptors, through radioligand binding studies, has been performed, until now, by using low-affinity and non-selective antagonists like 1,3-dipropyl-8 cyclopentylxanthine ([(3)H]DPCPX),(4-(2-[7-amino-2-(2-furyl)-[1,2,4]triazolo-[2,3 a][1,3,5]triazin-5-ylamino]ethyl)-phenol ([(3)H]ZM 241385) and 3-(3,4 aminobenzyl)-8-(4-oxyacetate)phenyl-1-propyl-xanthine ([(125)I]ABOPX). Recently, high-affinity radioligands for A(2B) receptors, [N-(4-cyanophenyl)-2-[4-(2,3,6,7 tetrahydro-2,6-dioxo-1,3-dipropyl-1H-purin-8-yl)-phenoxy]acetamide ([(3)H]MRS 1754), N-(2-(2-Phenyl-6-[4-(2,2,3,3-tetratritrio-3-phenylpropyl)-piperazine-1 carbonyl]-7H-pyrrolo[2,3-d]pyrimidin-4-ylamino)-ethyl)-acetamide ([(3)H]OSIP339391) and N-benzo[1,3]dioxol-5-yl-2-[5-(1,3-dipropyl-2,6-dioxo 2,3,6,7-tetrahydro-1H-purin-8-yl)-1-methyl-1H-pyrazol-3-yloxy]-acetamide] ([(3)H]MRE 2029F20), have been introduced. This minireview offers an overview of these recently developed radioligands and the most important applications of drugs towards A(2B) receptors. PMID- 18404461 TI - Molecular modelling study of 2-phenylethynyladenosine (PEAdo) derivatives as highly selective A3 adenosine receptor ligands. AB - A series of 2-phenylethynyladenosine (PEAdo) derivatives substituted in the N6- and 4'-position was synthesised and the new derivatives were tested at the four human adenosine receptors stably transfected into Chinese hamster ovary (CHO) cells, using radioligand binding studies (A1, A(2A), A3) or adenylyl cyclase activity assay (A(2B)). Binding studies showed that the presence of a phenyl ethynyl group in the 2 position of adenosine favoured the interaction with A3 receptors, resulting in compounds endowed with high affinity and selectivity for the A3 subtype. Additional substitution of the N6- and 4'-position increases both A3 affinity and selectivity. The results showed that the new compounds have a good affinity for the A3 receptor and in particular, the N6-methoxy-2 phenylethynyl-5'-N-methylcarboxamidoadenosine, with a K(i) at A3 of 1.9 nM and a selectivity A1/A3 and A(2A)/A3 of 4,800- and 8,600-fold, respectively. Therefore, it is one of the most potent and selective agonists at the human A3 adenosine receptor subtype reported so far. Furthermore, functional assays of inhibition of 10 MUM forskolin-stimulated cAMP production via the adenosine A3 receptor revealed that the new trisubstituted adenosine derivatives behave as full agonist of this receptor subtype. Docking analysis of these compounds was performed at a homology model of the human A3 receptor based on the bovine rhodopsin crystal structure as template, and the results are in accordance with the biological data. PMID- 18404462 TI - Roles of P2 receptors in glial cells: focus on astrocytes. AB - Central nervous system glial cells release and respond to nucleotides under both physiological and pathological conditions, suggesting that these molecules play key roles in both normal brain function and in repair after damage. In particular, ATP released from astrocytes activates P2 receptors on astrocytes and other brain cells, allowing a form of homotypic and heterotypic signalling, which also involves microglia, neurons and oligodendrocytes. Multiple P2X and P2Y receptors are expressed by both astrocytes and microglia; however, these receptors are differentially recruited by nucleotides, depending upon specific pathophysiological conditions, and also mediate the long-term trophic changes of these cells during inflammatory gliosis. In astrocytes, P2-receptor-induced gliosis occurs via activation of the extracellular-regulated kinases (ERK) and protein kinase B/Akt pathways and involves induction of inflammatory and anti inflammatory genes, cyclins, adhesion and antiapoptotic molecules. While astrocytic P2Y1 and P2Y(2,4) are primarily involved in short-term calcium dependent signalling, multiple P2 receptor subtypes seem to cooperate to astrocytic long-term changes. Conversely, in microglia, exposure to inflammatory and immunological stimuli results in differential functional changes of distinct P2 receptors, suggesting highly specific roles in acquisition of the activated phenotype. We believe that nucleotide-induced activation of astrocytes and microglia may originally start as a defence mechanism to protect neurons from cytotoxic and ischaemic insults; dysregulation of this process in chronic inflammatory diseases eventually results in neuronal cell damage and loss. On this basis, full elucidation of the specific roles of P2 receptors in these cells may help exploit the beneficial neuroprotective features of activated glia while attenuating their harmful properties and thus provide the basis for novel neuroprotective strategies that specifically target the purinergic system. PMID- 18404463 TI - Different patterns of Ca2+ signals are induced by low compared to high concentrations of P2Y agonists in microglia. AB - Brain-resident macrophages (microglia) are key cellular elements in the preservation of tissue integrity. On the other hand, they can also contribute to the development of pathological events by causing an extensive and inappropriate inflammatory response. A growing number of reports indicate the involvement of nucleotides in the control of microglial functions. With this study on P2Y receptors in rat microglia, we want to contribute to the definition of their expression profile and to the characterisation of their signalling mechanisms leading to Ca2+ movements. Endogenous nucleotides, when applied at a concentration of 100 muM, elicited robust Ca2+ transients, thanks to a panel of metabotropic receptors comprising mainly P2Y2, P2Y6 and P2Y12 subtypes. The involvement of P2Y12 receptors in Ca2+ responses induced by adenine nucleotides was confirmed by the pharmacological and pertussis toxin sensitivity of the response induced by adenosine diphosphate (ADP). Beside the G protein involved, Gi and Gq respectively, adenine and uracil nucleotides differed also for induction by the latter of a capacitative Ca2+ plateau. Moreover, when applied at low (sub-micromolar) concentrations with a long-lasting challenge, uracil nucleotides elicited oscillatory Ca2+ changes with low frequency of occurrence ( inosine + Pi), and the phosphate transfer from a nucleoside monophosphate donor to the 5' position of a nucleoside acceptor (e.g., IMP + guanosine --> inosine + GMP). The enzyme protein functions through the formation of a covalent phosphoenzyme intermediate, followed by the phosphate transfer either to water (phosphatase activity) or to a nucleoside (phosphotransferase activity). It has been proposed that cN-II regulates the intracellular concentration of IMP and GMP and the production of uric acid. The enzyme might also have a potential therapeutic importance, since it can phosphorylate some anti-tumoral and antiviral nucleoside analogues that are not substrates of known kinases. In this review we summarise our recent studies on the structure, regulation and function of cN-II. Via a site-directed mutagenesis approach, we have identified the amino acids involved in the catalytic mechanism and proposed a structural model of the active site. A series of in vitro studies suggests that cN-II might contribute to the regulation of 5-phosphoribosyl-1 pyrophosphate (PRPP) level, through the so-called oxypurine cycle, and in the production of intracellular adenosine, formed by ATP degradation. PMID- 18404471 TI - Editorial. PMID- 18404472 TI - The calcium activated nucleotidases: A diverse family of soluble and membrane associated nucleotide hydrolyzing enzymes. AB - It has long been known that the salivary glands of hematophagous (blood-feeding) arthropods secrete soluble apyrases, which are potent nucleotide hydrolyzing enzymes capable of hydrolyzing extracellular ATP and ADP, the latter being a major agonist contributing to platelet aggregation. Only recently, however, has the identification of proteins homologous to these apyrases been reported in non blood-feeding organisms such as rodents and humans. In this review, we present an overview of the diverse family of apyrases first described in the blood-feeding arthropods, including the identification and characterization of the soluble and membrane-bound vertebrate enzymes homologous to these arthropod apyrases. We also describe the enzymatic properties and nucleotide specificities of the expressed enzymes, and insights gained into the structure and function of this calcium activated nucleotidase (CAN) family from biophysical, mutagenesis and crystallography studies. The potential therapeutic value of these proteins is also discussed. PMID- 18404473 TI - Alkaline Phosphatases : Structure, substrate specificity and functional relatedness to other members of a large superfamily of enzymes. AB - Our knowledge of the structure and function of alkaline phosphatases has increased greatly in recent years. The crystal structure of the human placental isozyme has enabled us to probe salient features of the mammalian enzymes that differ from those of the bacterial enzymes. The availability of knockout mice deficient in each of the murine alkaline phosphatase isozymes has also given deep insights into their in vivo role. This has been particularly true for probing the biological role of bone alkaline phosphatase during skeletal mineralization. Due to space constraints this mini-review focuses exclusively on structural and functional features of mammalian alkaline phosphatases as identified by crystallography and probed by site-directed mutagenesis and kinetic analysis. An emphasis is also placed on the substrate specificity of alkaline phosphatases, their catalytic properties as phosphohydrolases as well as phosphodiesterases and their structural and functional relatedness to a large superfamily of enzymes that includes nucleotide pyrophosphatase/phosphodiesterase. PMID- 18404474 TI - Ecto-5'-nucleotidase: Structure function relationships. AB - Ecto-5'-nucleotidase (ecto-5'-NT) is attached via a GPI anchor to the extracellular membrane, where it hydrolyses AMP to adenosine and phosphate. Related 5'-nucleotidases exist in bacteria, where they are exported into the periplasmic space. X-ray structures of the 5'-nucleotidase from E. coli showed that the enzyme consists of two domains. The N-terminal domain coordinates two catalytic divalent metal ions, whereas the C-terminal domain provides the substrate specificity pocket for the nucleotides. Thus, the substrate binds at the interface of the two domains. Here, the currently available structural information on ecto-5'-NT is reviewed in relation to the catalytic properties and enzyme function. PMID- 18404475 TI - Physiological roles for ecto-5'-nucleotidase (CD73). AB - Nucleotides and nucleosides influence nearly every aspect of physiology and pathophysiology. Extracellular nucleotides are metabolized through regulated phosphohydrolysis by a series of ecto-nucleotidases. The formation of extracellular adenosine from adenosine 5'-monophosphate is accomplished primarily through ecto-5'-nucleotidase (CD73), a glycosyl phosphatidylinositol-linked membrane protein found on the surface of a variety of cell types. Recent in vivo studies implicating CD73 in a number of tissue protective mechanisms have provided new insight into its regulation and function and have generated considerable interest. Here, we review contributions of CD73 to cell and tissue stress responses, with a particular emphasis on physiologic responses to regulated CD73 expression and function, as well as new findings utilizing Cd73 deficient animals. PMID- 18404476 TI - Modulation of purinergic signaling by NPP-type ectophosphodiesterases. AB - Extracellular nucleotides can elicit a wide array of cellular responses by binding to specific purinergic receptors. The level of ectonucleotides is dynamically controlled by their release from cells, synthesis by ectonucleoside diphosphokinases and ectoadenylate kinases, and hydrolysis by ectonucleotidases. One of the four structurally unrelated families of ectonucleotidases is represented by the NPP-type ectophosphodiesterases. Three of the seven members of the NPP family, namely NPP1-3, are known to hydrolyze nucleotides. The enzymatic action of NPP1-3 (in)directly results in the termination of nucleotide signaling, the salvage of nucleotides and/or the generation of new messengers like ADP, adenosine or pyrophosphate. NPP2 is unique in that it hydrolyzes both nucleotides and lysophospholipids and, thereby, generates products that could synergistically promote cell motility. We review here the enzymatic properties of NPPs and analyze current evidence that links their nucleotide-hydrolyzing capability to epithelial and neural functions, the immune response and cell motility. PMID- 18404478 TI - CD39, NTPDase 1, is attached to the plasma membrane by two transmembrane domains. Why? AB - Since the identification of CD39 and other members of the e-NTPDase (ecto nucleoside triphosphate diphosphohydrolase) family as the primary enzymes responsible for cell surface nucleotide hydrolysis, one of their most intriguing features has been their unusual topology. The active site lies in the large extracellular region, but instead of being anchored in the membrane by a single transmembrane domain or lipid link like other ectoenzymes, CD39 has two transmembrane domains, one at each end. In this review we discuss evidence that the structure and dynamics of the transmembrane helices are intricately connected to enzymatic function. Removal of either or both transmembrane domains or disruption of their native state by detergent solubilization reduces activity by 90%, indicating that native function requires both transmembrane domains to be present and in the membrane. Enzymatic and mutational analysis of the native and truncated forms has shown that the active site can exist in distinct functional states characterized by different total activities, substrate specificities, hydrolysis mechanisms, and intermediate ADP release during ATP hydrolysis, depending on the state of the transmembrane domains. Disulfide crosslinking of cysteines introduced within the transmembrane helices revealed that they interact within and between molecules, in particular near the extracellular domain, and that activity depends on their organization. Both helices exhibit a high degree of rotational mobility, and the ability to undergo dynamic motions is required for activity and regulated by substrate binding. Recent reports suggest that membrane composition can regulate NTPDase activity. We propose that mechanical bilayer properties, potentially elasticity, might regulate CD39 by altering the balance between stability and mobility of its transmembrane domains. PMID- 18404477 TI - Physiologic and pathologic functions of the NPP nucleotide pyrophosphatase/phosphodiesterase family focusing on NPP1 in calcification. AB - The catabolism of ATP and other nucleotides participates partly in the important function of nucleotide salvage by activated cells and also in removal or de novo generation of compounds including ATP, ADP, and adenosine that stimulate purinergic signaling. Seven nucleotide pyrophosphatase/phosphodiesterase NPP family members have been identified to date. These isoenzymes, related by up conservation of catalytic domains and certain other modular domains, exert generally non-redundant functions via distinctions in substrates and/or cellular localization. But they share the capacity to hydrolyze phosphodiester or pyrophosphate bonds, though generally acting on distinct substrates that include nucleoside triphosphates, lysophospholipids and choline phosphate esters. PP(i) generation from nucleoside triphosphates, catalyzed by NPP1 in tissues including cartilage, bone, and artery media smooth muscle cells, supports normal tissue extracellular PP(i) levels. Balance in PP(i) generation relative to PP(i) degradation by pyrophosphatases holds extracellular PP(i) levels in check. Moreover, physiologic levels of extracellular PP(i) suppress hydroxyapatite crystal growth, but concurrently providing a reservoir for generation of pro mineralizing P(i). Extracellular PP(i) levels must be supported by cells in mineralization-competent tissues to prevent pathologic calcification. This support mechanism becomes dysregulated in aging cartilage, where extracellular PP(i) excess, mediated in part by upregulated NPP1 expression stimulates calcification. PP(i) generated by NPP1modulates not only hydroxyapatite crystal growth but also chondrogenesis and expression of the mineralization regulator osteopontin. This review pays particular attention to the role of NPP1-catalyzed PP(i) generation in the pathogenesis of certain disorders associated with pathologic calcification. PMID- 18404479 TI - E-NTPDases in human airways: Regulation and relevance for chronic lung diseases. AB - Chronic obstructive lung diseases are characterized by the inability to prevent bacterial infection and a gradual loss of lung function caused by recurrent inflammatory responses. In the past decade, numerous studies have demonstrated the importance of nucleotide-mediated bacterial clearance. Their interaction with P2 receptors on airway epithelia provides a rapid 'on-and-off' signal stimulating mucus secretion, cilia beating activity and surface hydration. On the other hand, abnormally high ATP levels resulting from damaged epithelia and bacterial lysis may cause lung edema and exacerbate inflammatory responses. Airway ATP concentrations are regulated by ecto nucleoside triphosphate diphosphohydrolases (E-NTPDases) which are expressed on the mucosal surface and catalyze the sequential dephosphorylation of nucleoside triphosphates to nucleoside monophosphates (ATP --> ADP --> AMP). The common bacterial product, Pseudomonas aeruginosa lipopolysaccharide (LPS), induces an acute reduction in azide sensitive E-NTPDase activities, followed by a sustained increase in activity as well as NTPDase 1 and NTPDase 3 expression. Accordingly, chronic lung diseases, including cystic fibrosis (CF) and primary ciliary dyskinesia, are characterized by higher rates of nucleotide elimination, azide-sensitive E-NTPDase activities and expression. This review integrates the biphasic regulation of airway E NTPDases with the function of purine signaling in lung diseases. During acute insults, a transient reduction in E-NTPDase activities may be beneficial to stimulate ATP-mediated bacterial clearance. In chronic lung diseases, elevating E NTPDase activities may represent an attempt to prevent P2 receptor desensitization and nucleotide-mediated lung damage. PMID- 18404481 TI - The CD38/CD157 mammalian gene family: An evolutionary paradigm for other leukocyte surface enzymes. AB - Human CD38 is the mammalian prototype of a family of phylogenetically conserved proteins which share structural similarities and enzymatic activities involved in the production of an intracellular second messenger with calcium mobilizing effects. Engagement of CD38 by agonistic monoclonal antibodies and the CD31 ligand initiates a cytoplasmic signaling cascade involving tyrosine phosphorylation of the proto-oncogene c-cbl and of the extracellular regulated kinase 1 of 2 complex. Further requirements for signal transduction include a privileged localization within the cholesterol-rich areas of the plasma membrane and physical association with specialized surface receptors. CD38-mediated signals are crucial in heterotypic cell adhesion and migration as well as in the activation of proliferation/survival programs by normal and neoplastic cells. Here we review the most recent literature on this complex topic and attempt to formulate a single model reconciling the enzymatic and receptorial activities of CD38. PMID- 18404482 TI - Plant responses to extracellular nucleotides: Cellular processes and biological effects. AB - Higher plants exhibit cellular responsiveness to the exogenous application of purine nucleotides in a manner consistent with a cell-cell signaling function for these molecules. Like animals, plants respond to extracellular ATP, ADP, and stable analogues (e.g., ATPgammaS and ADPbetaS) by increasing the cytoplasmic concentration of calcium. Agonist substrate specificity and concentration dependency suggest receptor mediation of these events, and, although the identity of the plant receptor is currently unknown, pharmacological analysis points to the involvement of a plasma membrane-localized calcium channel. Extracellular ATP can also induce the production of reactive oxygen species and stimulate an increase in the mRNA levels of a number of stress- and calcium-regulated genes, suggesting a role for nucleotide-based signaling in plant wound and defense responses. Furthermore, the growth and development of plants can also be altered by the application of external ATP. Recent studies are only beginning to uncover the complexities of plant signaling networks activated in response to extracellular ATP and how these might interact to affect plant physiological processes. PMID- 18404480 TI - The E-NTPDase family of ectonucleotidases: Structure function relationships and pathophysiological significance. AB - Ectonucleotidases are ectoenzymes that hydrolyze extracellular nucleotides to the respective nucleosides. Within the past decade, ectonucleotidases belonging to several enzyme families have been discovered, cloned and characterized. In this article, we specifically address the cell surface-located members of the ecto nucleoside triphosphate diphosphohydrolase (E-NTPDase/CD39) family (NTPDase1,2,3, and 8). The molecular identification of individual NTPDase subtypes, genetic engineering, mutational analyses, and the generation of subtype-specific antibodies have resulted in considerable insights into enzyme structure and function. These advances also allow definition of physiological and patho physiological implications of NTPDases in a considerable variety of tissues. Biological actions of NTPDases are a consequence (at least in part) of the regulated phosphohydrolytic activity on extracellular nucleotides and consequent effects on P2-receptor signaling. It further appears that the spatial and temporal expression of NTPDases by various cell types within the vasculature, the nervous tissues and other tissues impacts on several patho-physiological processes. Examples include acute effects on cellular metabolism, adhesion, activation and migration with other protracted impacts upon developmental responses, inclusive of cellular proliferation, differentiation and apoptosis, as seen with atherosclerosis, degenerative neurological diseases and immune rejection of transplanted organs and cells. Future clinical applications are expected to involve the development of new therapeutic strategies for transplantation and various inflammatory cardiovascular, gastrointestinal and neurological diseases. PMID- 18404485 TI - Increased 5-HT(3)-mediated signalling in pelvic afferent neurons from mice deficient in P2X(2) and/or P2X (3) receptor subunits. AB - Extracellular ATP and 5-hydroxytryptamine (5-HT) are both involved in visceral sensory pathways by interacting with P2X and 5-HT(3) receptors, respectively. We have investigated the changes in P2X and 5-HT(3)-mediated signalling in pelvic afferent neurons in mice deficient in P2X(2) and/or P2X(3) subunits by whole-cell recording of L(6)-S(2) dorsal root ganglion (DRG) neurons and by multi-unit recording of pelvic afferents of the colorectum. In wildtype DRG neurons, ATP evoked transient, sustained or mixed (biphasic) inward currents. Transient currents were absent in P2X(3) (-/-) neurons, whereas sustained currents were absent in P2X(2) (-/-) DRG neurons. Neither transient nor sustained currents were observed following application of ATP or alpha,beta-methylene ATP (alpha,beta meATP) in P2X(2)/P2X(3) (Dbl-/-) DRG neurons. 5-HT was found to induce a fast inward current in 63% of DRG neurons from wildtype mice, which was blocked by tropisetron, a 5-HT(3) receptor antagonist. The percentage of DRG neurons responding to 5-HT was significantly increased in P2X (2) (-/-), P2X(3) (-/-) and P2X(2)/P2X(3) (Dbl-/-) mice, and the amplitude of 5-HT response was significantly increased in P2X(2)/P2X(3) (Dbl-/-) mice. The pelvic afferent response to colorectal distension was attenuated in P2X(2)/P2X(3) (Dbl-/-) mice, but the response to serosal application of 5-HT was enhanced. Furthermore, tropisetron resulted in a greater reduction in pelvic afferent responses to colorectal distension in the P2X(2)/P2X(3) (Dbl-/-) preparations. These data suggest that P2X receptors containing the P2X(2) and/or P2X(3) subunits mediate purinergic activation of colorectal afferents and that 5-HT signalling in pelvic afferent neurons is up-regulated in mice lacking P2X(2) or P2X(3) receptor genes. This effect is more pronounced when both subunits are absent. PMID- 18404486 TI - Upregulation of P2Y2 receptors by retinoids in normal human epidermal keratinocytes. AB - Retinoids, vitamin A derivatives, are important regulators of the growth and differentiation of skin cells. Although retinoids are therapeutically used for several skin ailments, little is known about their effects on P2 receptors, known to be involved in various functions in the skin. DNA array analysis showed that treatment of normal human epidermal keratinocytes (NHEKs) with all-trans-retinoic acid (ATRA), an agonist to RAR (retinoic acid receptor), enhanced the expression of mRNA for the P2Y2 receptor, a metabotropic P2 receptor that is known to be involved in the proliferation of the epidermis. The expression of other P2 receptors in NHEKs was not affected by ATRA. ATRA increased the mRNA for the P2Y2 receptor in a concentration-dependent fashion (1 nM to 1 muM). Am80, a synthesized agonist to RAR, showed a similar enhancement, whereas 9-cis-retinoic acid (9-cisRA), an agonist to RXR (retinoid X receptor), enhanced P2Y2 gene expression to a lesser extent. Ca(2+) imaging analysis showed that ATRA also increased the function of P2Y2 receptors in NHEKs. Retinoids are known to enhance the turnover of the epidermis by increasing both proliferation and terminal differentiation. The DNA microarray analysis also revealed that ATRA upregulates various genes involved in the differentiation of NHEKs. Our present results suggest that retinoids, at least in part, exert their proliferative effects by upregulating P2Y2 receptors in NHEKs. This effect of retinoids may be closely related to their therapeutic effect against various ailments or aging events in skins such as over-keratinization, pigmentation and re-modeling. PMID- 18404484 TI - P2 receptors in atherosclerosis and postangioplasty restenosis. AB - Atherosclerosis is an immunoinflammatory process that involves complex interactions between the vessel wall and blood components and is thought to be initiated by endothelial dysfunction [1-3]. Extracellular nucleotides that are released from a variety of arterial and blood cells [4] can bind to P2 receptors and modulate proliferation and migration of smooth muscle cells (SMC), which is known to be involved in intimal hyperplasia that accompanies atherosclerosis and postangioplasty restenosis [5]. In addition, P2 receptors mediate many other functions, including platelet aggregation, leukocyte adherence, and arterial vasomotoricity. A direct pathological role of P2 receptors is reinforced by recent evidence showing that up-regulation and activation of P2Y(2) receptors in rabbit arteries mediates intimal hyperplasia [6]. In addition, up-regulation of functional P2Y receptors also has been demonstrated in the basilar artery of the rat double-hemorrhage model [7] and in coronary arteries of diabetic dyslipidemic pigs [8]. It has been proposed that up-regulation of P2Y receptors may be a potential diagnostic indicator for the early stages of atherosclerosis [9]. Therefore, particular effort must be made to understand the consequences of nucleotide release from cells in the cardiovascular system and the subsequent effects of P2 nucleotide receptor activation in blood vessels, which may reveal novel therapeutic strategies for atherosclerosis and restenosis after angioplasty. PMID- 18404487 TI - Stimulation of an alpha1-adrenergic receptor downregulates ecto-5' nucleotidase activity on the apical membrane of RPE cells. AB - The purines ATP and adenosine play an important role in the communication between the photoreceptors and the retinal pigment epithelium (RPE). While the RPE is known to release ATP into subretinal space, the source of extracellular adenosine is unclear. In other tissues, ecto-nucleotidases mediate the consecutive dephosphorylation of ATP to AMP, and AMP is converted to adenosine by ecto-5' nucleotidase (CD73). This study identifies ecto-5' nucleotidase on RPE cells and investigates modulation of enzyme activity. The RPE was the most active site of 5'AMP dephosphorylation in the posterior rat eye. The ecto-5' nucleotidase inhibitor alphabetamADP prevented the production adenosine by the apical membrane of the bovine RPE. Cultured human ARPE-19 cells expressed mRNA and protein for ecto-5' nucleotidase. The production of phosphate from 5'AMP by ARPE-19 cells was inhibited by alphabetamADP, but the ecto-alkaline phosphatase inhibitor levamisole had no effect. Degradation of 5'AMP was blocked by norepinephrine, epinephrine and phenylephrine, with inhibition by antagonists prazosin and corynanthine implicating the alpha1 adrenergic receptor. The block of enzyme activity by norepinephrine was rapid, occurring within 1 min, and was similar at both 4 and 37 degrees C, consistent with cleavage of the enzyme from its GPI anchor. HPLC measurements indicated norepinephrine reduced levels of adenosine in the bath. In the apical face of the bovine-RPE eyecup, norepinephrine reduced the production of phosphate from 5'AMP, suggesting that both receptor and enzyme face sub-retinal space. In conclusion, RPE cells express ecto-5' nucleotidase, with activity on the apical membrane, and stimulation of alpha-1 adrenergic receptors downregulates activity. As epinephrine is released at light onset, and adenosine can inhibit phagocytosis, the corresponding decrease in subretinal adenosine levels may contribute to the enhanced the phagocytosis of rod outer segments that occurs at this time. PMID- 18404483 TI - Integration of P2Y receptor-activated signal transduction pathways in G protein dependent signalling networks. AB - The role of nucleotides in intracellular energy provision and nucleic acid synthesis has been known for a long time. In the past decade, evidence has been presented that, in addition to these functions, nucleotides are also autocrine and paracrine messenger molecules that initiate and regulate a large number of biological processes. The actions of extracellular nucleotides are mediated by ionotropic P2X and metabotropic P2Y receptors, while hydrolysis by ecto-enzymes modulates the initial signal. An increasing number of studies have been performed to obtain information on the signal transduction pathways activated by nucleotide receptors. The development of specific and stable purinergic receptor agonists and antagonists with therapeutical potential largely contributed to the identification of receptors responsible for nucleotide-activated pathways. This article reviews the signal transduction pathways activated by P2Y receptors, the involved second messenger systems, GTPases and protein kinases, as well as recent findings concerning P2Y receptor signalling in C6 glioma cells. Besides vertical signal transduction, lateral cross-talks with pathways activated by other G protein-coupled receptors and growth factor receptors are discussed. PMID- 18404488 TI - P2X(5) and P2X(7) receptors in human warts and CIN-612 organotypic raft cultures of human papillomavirus infected keratinocytes. AB - Purinergic receptors, which bind adenosine 5'-triphosphate (ATP), are expressed on human cutaneous keratinocytes and in squamous cell carcinomas. Studies on normal human epidermis and primary keratinocyte cultures have suggested that P2X(5) receptors are likely to be involved in keratinocyte differentiation and P2X(7) receptors are likely to be part of the machinery of end stage terminal differentiation/apoptosis of keratinocytes. P2X(7) receptor agonists can significantly reduce primary keratinocyte cell numbers in culture. Human papillomaviruses are increasingly recognised as important human carcinogens in the development of non-melanoma skin cancers. In our study, immunohistochemical analysis for P2X(5) and P2X(7) receptors was performed on paraffin sections of normal human skin, warts, raft cultures of normal human keratinocytes and raft cultures of CIN 612 cells, a model of keratinocytes infected with human papillomavirus type 31. In warts there was up-regulation of the expression of P2X(5) receptors. A similar pattern was seen in the CIN 612 raft cultures. Both P2X(5) and P2X(7) receptors were found in the nuclei of koilocytes, abnormal keratinocytes characteristic of human papillomavirus infection. P2X(5) and P2X(7) receptors may provide a new focus for therapeutic research into treatments for warts because these receptors can induce cell differentiation and cell death. PMID- 18404489 TI - Adenosine 5'-triphosphate and its relationship with other mediators that activate pelvic nerve afferent neurons in the rat colorectum. AB - Evidence of a role for purinergic signalling in visceral afferents involving P2X(2), P2X(3) and P2Y(1) receptors exists, which appears to be important during inflammation. This study aimed to evaluate the degree of interaction between adenosine 5'-triphosphate (ATP) and other mediators that activate sensory nerves in the colorectum. Recordings from pelvic nerve afferents were made during application of agents to the in-vitro colorectal preparation. Analysis allowed calculation of single unit activity. When applied individually, bradykinin (78%) and 5-hydoxytryptamine (77%) activated the greatest number of neurons, followed by substance P, protons, ATP and capsaicin. Prostaglandin E(2) stimulated the least number (54%) and had a longer latency. Seventy-seven percent of all units studied either responded to both ATP and capsaicin or to neither, giving the greatest degree of activity correlation. Five percent of units were activated by all seven agents and no units were activated by a single agent alone. 5 hydroxytryptamine, capsaicin and protons, when co-applied with ATP, increased pelvic nerve activity to a greater degree than the sum of the individual responses. It is concluded that ATP activates pelvic nerve afferents and acts synergistically with protons, capsaicin and 5-hydroxytryptamine. The pattern of neuronal activation suggests that visceral afferents are polymodal but the receptor expression on their terminals is variable. PMID- 18404490 TI - Kinetics of ATP release following compression injury of a peripheral nerve trunk. AB - Compression and/or contusion of a peripheral nerve trunk can result in painful sensations. It is possible that release of ATP into the extracellular space may contribute to this symptom. In the present study, we used real-time measurements of ATP-induced bioluminescence together with electrophysiological recordings of compound action potentials to follow changes in the extracellular ATP concentration of isolated rat spinal roots exposed to mechanical stimuli. Nerve compression for about 8 s resulted in an immediate release of ATP into the extracellular space and in a decrease in the amplitude of compound action potentials. On average, a rise in ATP to 60 nM was observed when nerve compression blocked 50% of the myelinated axons. After the compression, the extracellular concentration of ATP returned to the resting level within a few minutes. The importance of ecto-nucleotidases for the recovery period was determined by exposure of isolated spinal roots to high concentrations of ATP and by use of inhibitors of ecto-nucleotidases. It was observed that spinal roots have a high capacity for ATP hydrolysis which is only partially blocked by betagamma-methylene ATP and ARL 67156. In conclusion, acute nerve compression produces an increase in the extracellular concentration of ATP and of its metabolites which may be sufficient for activation of purinergic P2 and/or P1 receptors on axons of nociceptive afferent neurons. PMID- 18404491 TI - Signaling pathways downstream of P2 receptors in human neutrophils. AB - Extracellular nucleotides stimulate human neutrophils by activating the purinergic P2Y(2) receptor. However, it is not completely understood which types of G proteins are activated downstream of this P2 receptor subtype. We investigated the G-protein coupling to P2Y(2) receptors and several subsequent signaling events. Treatment of neutrophils with pertussis toxin (PTX), a Gi protein inhibitor, caused only approximately 75% loss of nucleotide-induced Ca(2+) mobilization indicating that nucleotides cause Ca(2+) mobilization both through Gi-dependent and Gi-independent pathways. However, the PLC inhibitor U73122 almost completely inhibited Ca(2+) mobilization in both nucleotide- and fMLP-stimulated neutrophils, strongly supporting the view that both the PTX sensitive and the PTX-insensitive mechanism of Ca(2+) increase require activation of PLC. We investigated the dependence of ERK phosphorylation on the Gi pathway. Treatment of neutrophils with PTX caused almost complete inhibition of ERK phosphorylation in nucleotide or fMLP activated neutrophils. U73122 caused inhibition of nucleotide- or fMLP-stimulated ERK phosphorylation, suggesting that although pertussis toxin-insensitive pathways cause measurable Ca(2+) mobilization, they are not sufficient for causing ERK phosphorylation. Since PLC activation leads to intracellular Ca(2+) increase and PKC activation, we investigated if these intracellular events are necessary for ERK phosphorylation. Exposure of cells to the Ca(2+) chelator BAPTA had no effect on nucleotide- or fMLP-induced ERK phosphorylation. However, the PKC inhibitor GF109203X was able to almost completely inhibit nucleotide- or fMLP-induced ERK phosphorylation. We conclude that the P2Y(2) receptor can cause Ca(2+) mobilization through a PTX insensitive but PLC-dependent pathway and ERK phosphorylation is highly dependent on activation of the Gi proteins. PMID- 18404492 TI - Excitatory effect of ATP on rat area postrema neurons. AB - ATP-induced inward currents and increases in the cytosolic Ca(2+) concentration ([Ca](in)) were investigated in neurons acutely dissociated from rat area postrema using whole-cell patch-clamp recordings and fura-2 microfluorometry, respectively. The ATP-induced current (I (ATP)) and [Ca](in) increases were mimicked by 2-methylthio-ATP and ATP-gammaS, and were inhibited by P2X receptor (P2XR) antagonists. The current-voltage relationship of the I (ATP) exhibited a strong inward rectification, and the amplitude of the I (ATP) was concentration dependent. The I (ATP) was markedly reduced in the absence of external Na(+), and the addition of Ca(2+) to Na(+)-free saline increased the I (ATP). ATP did not increase [Ca](in) in the absence of external Ca(2+), and Ca(2+) channel antagonists partially inhibited the ATP-induced [Ca](in) increase, indicating that ATP increases [Ca](in) by Ca(2+) influx through both P2XR channels and voltage-dependent Ca(2+) channels. There was a negative interaction between P2XR- and nicotinic ACh receptor (nAChR)-channels, which depended on the amplitude and direction of current flow through either channel. Current occlusion was observed at V (h)s between -70 and -10 mV when the I (ATP) and ACh-induced current (I (ACh)) were inward, but no occlusion was observed when these currents were outward at a V (h) of +40 mV. The I (ATP) was not inhibited by co-application of ACh when the I (ACh) was markedly decreased either by removal of permeant cations, by setting V (h) close to the equilibrium potential of I (ACh), or by the addition of d-tubocurarine or serotonin. These results suggest that the inhibitory interaction is attributable to inward current flow of cations through the activated P2XR- and nAChR-channels. PMID- 18404493 TI - Characterization of human and rodent native and recombinant adenosine A(2B) receptors by radioligand binding studies. AB - Adenosine A(2B) receptors of native human and rodent cell lines were investigated using [(3)H]PSB-298 [(8-{4-[2-(2-hydroxyethylamino)-2-oxoethoxy]phenyl}-1 propylxanthine] in radioligand binding studies. [(3)H]PSB-298 showed saturable and reversible binding. It exhibited a K(D) value of 60 +/- 1 nM and limited capacity (B(max) = 3.511 fmol per milligram protein) at recombinant human adenosine A(2B) receptors expressed in human embryonic kidney cells (HEK-293). The addition of sodium chloride (100 mM) led to a threefold increase in the number of binding sites recognized by the radioligand. The curve of the agonist 5'-N-ethylcarboxamidoadenosine (NECA) was shifted to the right in the presence of NaCl, while the curve of the antagonist PSB-298 was shifted to the left, indicating that PSB-298 may be an inverse agonist at A(2B) receptors. Adenosine A(2B) receptors were shown to be the major adenosine A(2) receptor subtype on the mouse neuroblastoma x rat glioma hybrid cell line NG108-15 cells. Binding studies at rat INS-1 cells (insulin secreting cell line) demonstrated that [(3)H]PSB-298 is a selective radioligand for adenosine A(2B) binding sites in this cell line. PMID- 18404495 TI - ATP receptors in pain sensation: Involvement of spinal microglia and P2X(4) receptors. AB - There is abundant evidence that extracellular ATP and other nucleotides have an important role in pain signaling at both the periphery and in the CNS. At first, it was thought that ATP was simply involved in acute pain, since ATP is released from damaged cells and excites directly primary sensory neurons by activating their receptors. However, neither blocking P2X/Y receptors pharmacologically nor suppressing the expression of P2X/Y receptors molecularly in sensory neurons or in the spinal cord had an effect on acute physiological pain. The focus of attention now is on the possibility that endogenous ATP and its receptor system might be activated in pathological pain states, particularly in neuropathic pain. Neuropathic pain is often a consequence of nerve injury through surgery, bone compression, diabetes or infection. This type of pain can be so severe that even light touching can be intensely painful; unfortunately, this state is generally resistant to currently available treatments. An important advance in our understanding of the mechanisms involved in neuropathic pain has been made by a recent work demonstrating the crucial role of ATP receptors (i.e., P2X(3) and P2X(4) receptors). In this review, we summarize the role of ATP receptors, particularly the P2X(4) receptor, in neuropathic pain. The expression of P2X(4) receptors in the spinal cord is enhanced in spinal microglia after peripheral nerve injury, and blocking pharmacologically and suppressing molecularly P2X(4) receptors produce a reduction of the neuropathic pain behaviour. Understanding the key roles of ATP receptors including P2X(4) receptors may lead to new strategies for the management of neuropathic pain. PMID- 18404496 TI - Responses of the ciliates Tetrahymena and Paramecium to external ATP and GTP. AB - The unicellular ciliates Paramecium and Tetrahymena are the simplest eukaryotic cells to show reliable depolarizing responses to micromolar concentrations of external ATP and GTP. Their simplicity allows for combined analysis of swimming behavior, electrophysiology, receptor binding, behavioral mutant and drug screens as well as molecular genetic approaches such as RNAi and gene knockouts experiments. ATP and GTP are depolarizing chemorepellents in both ciliates, producing measurable receptor potentials and Ca(2+)-based action potentials that are correlated with jerking behaviors called avoiding reactions (AR). GTP also causes repetitive continuous ciliary reversals (CCR) and oscillating plateau depolarizations in Paramecium. Both ciliates show high affinity, saturable external binding of (32)P-GTP and (32)P-ATP but GTP does not compete for ATP binding and vice versa. Chemosensory adaptation occurs after continued exposure (15 min) to these ligands, producing a loss of external binding and forward swimming. However, cells adapted to ATP still bind and respond to GTP and GTP adapted cells still bind and respond to ATP. This, combined with pharmacological analyses, suggests that there are two separate receptor systems: A metabotropic ATP receptor pathway and a different, novel GTP receptor pathway. A Paramecium mutant (ginA) lacks the GTP-induced oscillating depolarizations but does show AR in GTP, unveiling isolated GTP-receptor potentials for study. An ecto-ATPase is also present that may be involved in inactivation of ATP and GTP signals. Gene knockout experiments are currently underway to determine the roles of the ecto ATPase and a putative 7-transmembrane spanning receptor in these responses. PMID- 18404498 TI - Release and extracellular metabolism of ATP by ecto-nucleotidase eNTPDase 1-3 in hypothalamic and pituitary cells. AB - Hypothalamic and pituitary cells express G protein-coupled adenosine and P2Y receptors and cation-conducting P2X receptor-channels, suggesting that extracellular ATP and other nucleotides may function as autocrine and/or paracrine signaling factors in these cells. Consistent with this hypothesis, we show that cultured normal and immortalized pituitary and hypothalamic cells release ATP under resting conditions. RT-PCR analysis also revealed the presence of transcripts for ecto-nucleotidase eNTPDase 1-3 in these cells. These enzymes were functional as documented by degradation of endogenously released and exogenously added ATP. Blocking the activity of eNTPDases by ARL67156 led to an increase in ATP release in perifused pituitary cells and inhibition of degradation of extracellularly added ATP. Furthermore, the addition of apyrase, a soluble ecto-nucleotidase, and the expression of recombinant mouse eNTPDase-2, enhanced degradation of both endogenously released and exogenously added ATP. The released ATP by resting hypothalamic cells was sufficient to activate and desensitize high-affinity recombinant P2X receptors, whereas facilitation of ATP metabolism by the addition of apyrase protected their desensitization. These results indicate that colocalization of ATP release sites and ecto-nucleotidase activity at the plasma membrane of hypothalamic and pituitary cells provides an effective mechanism for the operation of nucleotides as extracellular signaling molecules. PMID- 18404499 TI - The P2Y(1) receptor is involved in the maintenance of glucose homeostasis and in insulin secretion in mice. AB - Pancreatic beta cells express several P2 receptors including P2Y(1) and the modulation of insulin secretion by extracellular nucleotides has suggested that these receptors may contribute to the regulation of glucose homeostasis. To determine whether the P2Y(1) receptor is involved in this process, we performed studies in P2Y(1)-/- mice. In baseline conditions, P2Y(1)-/- mice exhibited a 15% increase in glycemia and a 40% increase in insulinemia, associated with a 10% increase in body weight, pointing to a role of the P2Y(1) receptor in the control of glucose metabolism. Dynamic experiments further showed that P2Y(1)-/- mice exhibited a tendency to glucose intolerance. These features were associated with a decrease in the plasma levels of free fatty acid and triglycerides. When fed a lipids and sucrose enriched diet for 15 weeks, the two genotypes no longer displayed any significant differences. To determine whether the P2Y(1) receptor was directly involved in the control of insulin secretion, experiments were carried out in isolated Langerhans islets. In the presence of high concentrations of glucose, insulin secretion was significantly greater in islets from P2Y(1)-/- mice. Altogether, these results show that the P2Y(1) receptor plays a physiological role in the maintenance of glucose homeostasis at least in part by regulating insulin secretion. PMID- 18404500 TI - Cerebellar astrocytes co-express several ADP receptors. Presence of functional P2Y(13)-like receptors. AB - Astrocytes exhibit a form of excitability based on variations of intracellular Ca(2+) concentration in response to various stimuli, including ADP, ATP, UTP and dinucleotides. Here, we investigate the presence of the recently cloned ADP sensitive receptors, P2Y(12) and P2Y(13) subtypes, which are negatively coupled to adenylate cyclase, in cerebellar astrocytes. We checked the effect of specific agonists, 2-methylthioadenosine diphosphate (2MeSADP) and ADP, on adenylate cyclase stimulation induced by isoproterenol. Both agonists significantly reduced the cAMP accumulation induced by isoproterenol. The inhibitory effect was concentration-dependent with IC(50) values of 46 +/- 13 and 23 +/- 14 nM for 2MeSADP and ADP, respectively. The experiments were carried out in the presence of MRS-2179, a specific antagonist of P2Y(1) receptor, to avoid any contribution of this receptor. Using fura-2 microfluorimetry we also proved that astrocytes responded to 2MeSADP stimulations with calcium responses in the absence and also in the presence of MRS-2179. Both effects, inhibition of adenylate cyclase and intracellular calcium mobilization, were not modified by 2MeSAMP, an antagonist of P2Y(12) receptor, suggesting that were mediated by P2Y(13)-like receptors. PMID- 18404502 TI - 2- and 8-alkynyl-9-ethyladenines: Synthesis and biological activity at human and rat adenosine receptors. AB - The synthesis of a series of 9-ethyladenine derivatives bearing alkynyl chains in 2- or 8-position was undertaken, based on the observation that replacement of the sugar moiety in adenosine derivatives with alkyl groups led to adenosine receptor antagonists. All the synthesized compounds were tested for their affinity at human and rat A(1), A(2A), and A(3) adenosine receptors in binding assays; the activity at the human A(2B) receptor was determined in adenylyl cyclase experiments. Biological data showed that the 2-alkynyl derivatives possess good affinity and are slightly selective for the human A(2A) receptor. The same compounds tested on the rat A(1) and A(2A) subtypes showed in general lower affinity for both receptors. On the other hand, the affinity of the 8-alkynyl derivatives at the human A(1), A(2A), and A(2B) receptors proved to be lower than that of the corresponding 2-alkynyl derivatives. On the contrary, the affinity of the same compounds for the human A(3) receptor was improved, resulting in A(3) selectivity. As in the case of the 2-alkynyl-substituted compounds, the 8-alkynyl derivatives showed decreased affinity for rat receptors. However, it is worthwhile to note that the 8-phenylethynyl-9-ethyladenine was the most active compound of the two series (K (i) in the nanomolar range) at both the human and rat A(3) subtype. Docking experiments of the 2- and 8-phenylethynyl-9 ethyladenines, at a rhodopsin-based homology model, gave a rational explanation of the preference of the human A(3) receptor for the 8-substituted compound. PMID- 18404501 TI - Guanosine stimulates neurite outgrowth in PC12 cells via activation of heme oxygenase and cyclic GMP. AB - Undifferentiated rat pheochromocytoma (PC12) cells extend neurites when cultured in the presence of nerve growth factor (NGF). Extracellular guanosine synergistically enhances NGF-dependent neurite outgrowth. We investigated the mechanism by which guanosine enhances NGF-dependent neurite outgrowth. Guanosine administration to PC12 cells significantly increased guanosine 3',5'-cyclic monophosphate (cGMP) within the first 24 h whereas addition of soluble guanylate cyclase (sGC) inhibitors abolished guanosine-induced enhancement of NGF-dependent neurite outgrowth. sGC may be activated either by nitric oxide (NO) or by carbon monoxide (CO). [Formula: see text]-Nitro-L-: arginine methyl ester (L-: NAME), a non-isozyme selective inhibitor of nitric oxide synthase (NOS), had no effect on neurite outgrowth induced by guanosine. Neither nNOS (the constitutive isoform), nor iNOS (the inducible isoform) were expressed in undifferentiated PC12 cells, or under these treatment conditions. These data imply that NO does not mediate the neuritogenic effect of guanosine. Zinc protoporphyrin-IX, an inhibitor of heme oxygenase (HO), reduced guanosine-dependent neurite outgrowth but did not attenuate the effect of NGF. The addition of guanosine plus NGF significantly increased the expression of HO-1, the inducible isozyme of HO, after 12 h. These data demonstrate that guanosine enhances NGF-dependent neurite outgrowth by first activating the constitutive isozyme HO-2, and then by inducing the expression of HO-1, the enzymes responsible for CO synthesis, thus stimulating sGC and increasing intracellular cGMP. PMID- 18404503 TI - Structure-activity relationships of dinucleotides: Potent and selective agonists of P2Y receptors. AB - Dinucleoside polyphosphates act as agonists on purinergic P2Y receptors to mediate a variety of cellular processes. Symmetrical, naturally occurring purine dinucleotides are found in most living cells and their actions are generally known. Unsymmetrical purine dinucleotides and all pyrimidine containing dinucleotides, however, are not as common and therefore their actions are not well understood. To carry out a thorough examination of the activities and specificities of these dinucleotides, a robust method of synthesis was developed to allow manipulation of either nucleoside of the dinucleotide as well as the phosphate chain lengths. Adenosine containing dinucleotides exhibit some level of activity on P2Y(1) while uridine containing dinucleotides have some level of agonist response on P2Y(2) and P2Y(6). The length of the linking phosphate chain determines a different specificity; diphosphates are most accurately mimicked by dinucleoside triphosphates and triphosphates most resemble dinucleoside tetraphosphates. The pharmacological activities and relative metabolic stabilities of these dinucleotides are reported with their potential therapeutic applications being discussed. PMID- 18404497 TI - Neuroprotection by adenosine in the brain: From A(1) receptor activation to A (2A) receptor blockade. AB - Adenosine is a neuromodulator that operates via the most abundant inhibitory adenosine A(1) receptors (A(1)Rs) and the less abundant, but widespread, facilitatory A(2A)Rs. It is commonly assumed that A(1)Rs play a key role in neuroprotection since they decrease glutamate release and hyperpolarize neurons. In fact, A(1)R activation at the onset of neuronal injury attenuates brain damage, whereas its blockade exacerbates damage in adult animals. However, there is a down-regulation of central A(1)Rs in chronic noxious situations. In contrast, A(2A)Rs are up-regulated in noxious brain conditions and their blockade confers robust brain neuroprotection in adult animals. The brain neuroprotective effect of A(2A)R antagonists is maintained in chronic noxious brain conditions without observable peripheral effects, thus justifying the interest of A(2A)R antagonists as novel protective agents in neurodegenerative diseases such as Parkinson's and Alzheimer's disease, ischemic brain damage and epilepsy. The greater interest of A(2A)R blockade compared to A(1)R activation does not mean that A(1)R activation is irrelevant for a neuroprotective strategy. In fact, it is proposed that coupling A(2A)R antagonists with strategies aimed at bursting the levels of extracellular adenosine (by inhibiting adenosine kinase) to activate A(1)Rs might constitute the more robust brain neuroprotective strategy based on the adenosine neuromodulatory system. This strategy should be useful in adult animals and especially in the elderly (where brain pathologies are prevalent) but is not valid for fetus or newborns where the impact of adenosine receptors on brain damage is different. PMID- 18404494 TI - Invited Lectures : Overviews Purinergic signalling: past, present and future. PMID- 18404504 TI - Comparative hydrolysis of P2 receptor agonists by NTPDases 1, 2, 3 and 8. AB - Nucleoside triphosphate diphosphohydrolases 1, 2, 3 and 8 (NTPDases 1, 2, 3 and 8) are the dominant ectonucleotidases and thereby expected to play important roles in nucleotide signaling. Distinct biochemical characteristics of individual NTPDases should allow them to regulate P2 receptor activation differentially. Therefore, the biochemical and kinetic properties of these enzymes were compared. NTPDases 1, 2, 3 and 8 efficiently hydrolyzed ATP and UTP with K (m) values in the micromolar range, indicating that they should terminate the effects exerted by these nucleotide agonists at P2X(1-7) and P2Y(2,4,11) receptors. Since NTPDase1 does not allow accumulation of ADP, it should terminate the activation of P2Y(1,12,13) receptors far more efficiently than the other NTPDases. In contrast, NTPDases 2, 3 and 8 are expected to promote the activation of ADP specific receptors, because in the presence of ATP they produce a sustained (NTPDase2) or transient (NTPDases 3 and 8) accumulation of ADP. Interestingly, all plasma membrane NTPDases dephosphorylate UTP with a significant accumulation of UDP, favoring P2Y(6) receptor activation. NTPDases differ in divalent cation and pH dependence, although all are active in the pH range of 7.0-8.5. Various NTPDases may also distinctly affect formation of extracellular adenosine and therefore adenosine receptor-mediated responses, since they generate different amounts of the substrate (AMP) and inhibitor (ADP) of ecto-5'-nucleotidase, the rate limiting enzyme in the production of adenosine. Taken together, these data indicate that plasma membrane NTPDases hydrolyze nucleotides in a distinctive manner and may therefore differentially regulate P2 and adenosine receptor signaling. PMID- 18404505 TI - Purinergic mechanism in the immune system: A signal of danger for dendritic cells. AB - There is increasing appreciation that injured or stressed cells release molecules endowed with the ability to modulate dendritic cell maturation. The role of these molecules is thought to be that of alerting the body of an impending danger, and initiate and shape the subsequent immune response. Nucleotides are perfectly suited for this task as they are easily released upon damage of the cell membrane, rapidly diffuse in the extracellular environment and ligate specific plasma membrane receptors expressed by dendritic cells and other mononuclear phagocytes. A better knowledge of the modulation of dendritic cell responses by extracellular nucleotides may provide novel routes to enhance the immune response and increase the efficacy of vaccination. PMID- 18404506 TI - Regulation of cell-to-cell communication mediated by astrocytic ATP in the CNS. AB - It has become apparent that glial cells, especially astrocytes, not merely supportive but are integrative, being able to receive inputs, assimilate information and send instructive chemical signals to other neighboring cells including neurons. At first, the excitatory neurotransmitter glutamate was found to be a major extracellular messenger that mediates these communications because it can be released from astrocytes in a Ca(2+)-dependent manner, diffused, and can stimulate extra-synaptic glutamate receptors in adjacent neurons, leading to a dynamic modification of synaptic transmission. However, recently extracellular ATP has come into the limelight as an important extracellular messenger for these communications. Astrocytes express various neurotransmitter receptors including P2 receptors, release ATP in response to various stimuli and respond to extracellular ATP to cause various physiological responses. The intercellular communication "Ca(2+) wave" in astrocytes was found to be mainly mediated by the release of ATP and the activation of P2 receptors, suggesting that ATP is a dominant "gliotransmitter" between astrocytes. Because neurons also express various P2 receptors and synapses are surrounded by astrocytes, astrocytic ATP could affect neuronal activities and even dynamically regulate synaptic transmission in adjacent neurons as if forming a "tripartite synapse". In this review, we summarize the role of astrocytic ATP, as compared with glutamate, in gliotransmission and synaptic transmission in neighboring cells, mainly focusing on the hippocampus. Dynamic communication between astrocytes and neurons mediated by ATP would be a key event in the processing or integration of information in the CNS. PMID- 18404507 TI - P2X(7) receptor: Death or life? AB - The P2X(7) plasma membrane receptor is an intriguing molecule that is endowed with the ability to kill cells, as well as to activate many responses and even stimulate proliferation. Here, the authors give an overview on the multiplicity and complexity of P2X(7)-mediated responses, discussing recent information on this receptor. Particular attention has been paid to early and late signs of apoptosis and necrosis linked to activation of the receptor and to the emerging field of P2X(7) function in carcinogenesis. PMID- 18404509 TI - P2 nucleotide receptors on C2C12 satellite cells. AB - In developing muscle cells environmental stimuli transmitted by purines binding to the specific receptors are crucial proliferation regulators. C2C12 myoblasts express numerous purinergic receptors representing both main classes: P2X and P2Y. Among P2Y receptors we have found the expression of P2Y(1), P2Y(2), P2Y(4), P2Y(6) and P2Y(12) family members while among P2X receptors P2X(4), P2X(5) and P2X(7) were discovered. We have been able to show that activation of those receptors is responsible for ERK class kinase activity, responsible for regulation of cell proliferation pathway. We have also demonstrated that this activity is calcium dependent suggesting Ca(2+) ions as secondary messenger between receptor and kinase regulatory system. More specifically, we do suspect that in C2C12 myoblasts calcium channels of P2X receptors, particularly P2X(5) play the main role in proliferation regulation. In further development of myoblasts into myotubes, when proliferation is gradually inhibited, the pattern of P2 receptors is changed. This phenomenon is followed by diminishing of the P2Y(2)-dependent Ca(2+) signaling, while the mRNA expression of P2Y(2) receptor reminds still on the high level. Moreover, P2X(2) receptor mRNA, absent in myoblasts appears in myotubes. These data show that differentiation of C2C12 cell line satellite myoblasts is accompanied by changes in P2 receptors expression pattern. PMID- 18404508 TI - Dynamic purine signaling and metabolism during neutrophil-endothelial interactions. AB - During episodes of hypoxia and inflammation, polymorphonuclear leukocytes (PMN) move into underlying tissues by initially passing between endothelial cells that line the inner surface of blood vessels (transendothelial migration, TEM). TEM creates the potential for disturbances in vascular barrier and concomitant loss of extravascular fluid and resultant edema. Recent studies have demonstrated a crucial role for nucleotide metabolism and nucleoside signaling during inflammation. These studies have implicated multiple adenine nucleotides as endogenous tissue protective mechanisms invivo. Here, we review the functional components of vascular barrier, identify strategies for increasing nucleotide generation and nucleoside signaling, and discuss potential therapeutic targets to regulate the vascular barrier during inflammation. PMID- 18404510 TI - Cloning and characterization of the ecto-nucleotidase NTPDase3 from rat brain: Predicted secondary structure and relation to other members of the E-NTPDase family and actin. AB - The protein family of ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDase family) contains multiple members that hydrolyze nucleoside 5'-triphosphates and nucleoside 5'-diphosphates with varying preference for the individual type of nucleotide. We report the cloning and functional expression of rat NTPDase3. The rat brain-derived cDNA has an open reading frame of 1590 bp encoding 529 amino acid residues, a calculated molecular mass of 59.1 kDa and predicted N- and C terminal hydrophobic sequences. It shares 94.3% and 81.7% amino acid identity with the mouse and human NTPDase3, respectively, and is more closely related to cell surface-located than to the intracellularly located members of the enzyme family. The NTPDase3 gene is allocated to chromosome 8q32 and organized into 11 exons. Rat NTPDase3 expressed in CHO cells hydrolyzed both nucleoside triphosphates and nucleoside diphosphates with hydrolysis ratios of ATP:ADP of 5:1 and UTP:UDP of 8:1. After addition of ATP, ADP is formed as an intermediate product that is further hydrolyzed to AMP. The enzyme is preferentially activated by Ca(2+) over Mg(2+) and reveals an alkaline pH optimum. Immunocytochemistry confirmed expression of heterologously expressed NTPDase3 to the surface of CHO cells. PC12 cells express endogenous surface-located NTPDase3. An immunoblot analysis detects NTPDase3 in all rat brain regions investigated. An alignment of the secondary structure domains of actin conserved within the actin/HSP70/sugar kinase superfamily to those of all members of the NTPDase family reveals apparent similarity. It infers that NTPDases share the two-domain structure with members of this enzyme superfamily. PMID- 18404511 TI - Identification of a tyrosine residue responsible for N-acetylimidazole-induced increase of activity of ecto-nucleoside triphosphate diphosphohydrolase 3. AB - Chemical modification in combination with site-directed mutagenesis was used to identify a tyrosine residue responsible for the increase in ecto-nucleoside triphosphate diphosphohydrolase 3 (NTPDase3) nucleotidase activity after acetylation with a tyrosine-selective reagent, N-acetylimidazole. The NTPDase3 ATPase activity is increased more than the ADPase activity by this reagent. Several fairly well conserved tyrosine residues (252, 255, and 262) that are located in or very near apyrase conserved region 4a (ACR4a) were mutated. These mutants were all active, but mutation of tyrosine 252 to either alanine or phenylalanine eliminated the activity increase observed after N-acetylimidazole treatment of the wild-type enzyme. This suggests that the acetylation of tyrosine 252 is responsible for the increased activity. Stabilization of quaternary structure has resulted in increased enzyme activities for the NTPDases. However, mutation of these three tyrosine residues did not result in global changes of tertiary or quaternary structure, as measured by Cibacron blue binding, chemical cross linking, and native gel electrophoretic analysis. Nevertheless, disruption of the oligomeric structure with the detergent Triton X-100 abolished the increase in activity induced by this reagent. In addition, mutations that abolished the N-acetylimidazole effect also attenuated the increases of enzyme activity observed after lectin and chemical cross-linking treatments, which were previously attributed to stabilization of the quaternary structure. Thus, we speculate that the acetylation of tyrosine 252 might induce a subtle conformational change in NTPDase3, resulting in the observed increase in activity. PMID- 18404512 TI - Tri-nucleotide receptors play a critical role in epithelial cell wound repair. AB - The cornea plays a major role in the refraction of light to the retina. Therefore, the integrity and transparency of the corneal epithelium are critical to vision. Following injury, a combination of rapid signal transduction events and long-term cell migration are essential for wound closure. We have demonstrated previously that injury resulted in the release of nucleotides that induce the propagation of a Ca(2+) wave to neighboring cells. This suggests that nucleotides and their receptors are critical components of wound healing. Epidermal growth factor (EGF) and integrins also have been shown to play a role in injury. In this study, we demonstrate that pretreatment of cells with ATP and UTP inhibited the immediate wound response, while BzATP, ADP, and UDP did not affect this response. Tri-nucleotide pretreatment also reduced the EGF induced Ca(2+) response. Additionally, lower EC(50) concentrations of ATP and UTP triggered migration of cells that was enhanced further with EGF and was inhibited by the tripeptide, RGD. Results indicate that the desensitization induced by ATP and UTP was specific. While ADP and UDP cause a homologous desensitization of their own signal, they did not cause an inhibition of the wound response nor does BzATP. Neither Ca(2+) wave propagation nor cell migration occurred in response to beta,gamma-MeATP. Together these results lead us to hypothesize that corneal epithelial wound repair is mediated by both P2Y(2) and P2Y(4) receptors. PMID- 18404513 TI - Editorial. PMID- 18404514 TI - First joint Italian-German purine club meeting "Progress in Purinergic Receptor Pharmacology and Function". PMID- 18404515 TI - Extracellular zinc and ATP-gated P2X receptor calcium entry channels: New zinc receptors as physiological sensors and therapeutic targets. AB - In this review, we focus on two attributes of P2X receptor channel function, one essential and one novel. First, we propose that P2X receptors are extracellular sensors as well as receptors and ion channels. In particular, the large extracellular domain (that comprises 70% of the molecular mass of the receptor channel protein) lends itself to be a cellular sensor. Moreover, its exquisite sensitivity to extracellular pH, ionic strength, and multiple ligands evokes the function of a sensor. Second, we propose that P2X receptors are extracellular zinc receptors as well as receptors for nucleotides. We provide novel data in multiple publications and illustrative data in this invited review to suggest that zinc triggers ATP-independent activation of P2X receptor channel function. In this light, P2X receptors are the cellular site of integration between autocrine and paracrine zinc signaling and autocrine and paracrine purinergic signaling. P2X receptors may sense changes in these ligands as well as in extracellular pH and ionic strength and transduce these sensations via calcium and/or sodium entry and changes in membrane potential. PMID- 18404517 TI - Cell cycle regulation of astrocytes by extracellular nucleotides and fibroblast growth factor-2. AB - Extracellular ATP enhances the mitogenic activity of fibroblast growth factor-2 (FGF2) in astrocytes, but the molecular mechanism underlying this synergistic interaction is not known. To determine whether the potentiating effect of extracellular ATP involves cell cycle control mechanisms, we have measured the expression of cyclins that are induced in different phases of the cell cycle in primary cultures of rat cortical astrocytes. We found that ATP potentiated the ability of FGF2 to stimulate expression of cyclin D1, a regulator of cell cycle entry, as well as cyclin A, a regulator of DNA replication. Because FGF2 and P2 purinergic receptors are coupled to extracellular signal regulated protein kinase (ERK), a key member of a signaling cascade that regulates proliferation, we also investigated the role of ERK in regulating cyclin expression induced by FGF2 and ATP. We found that the potentiating effect of ATP on cyclin expression was significantly reduced by U0126, an inhibitor of MEK, the upstream activator of ERK. P2 receptor agonist studies revealed that UTP enhanced FGF2-induced cyclin expression and mitogenesis whereas 2-methylthioADP was ineffective. By contrast, 2',3'-O-(4-benzoyl)-benzoyl-ATP markedly inhibited FGF2-induced mitogenesis. Consistent with opposing effects of P2Y and P2X receptors on mitogenesis, UTP stimulated a transient activation of ERK whereas BzATP stimulated a more sustained ERK signal. These findings suggest that signaling by P2Y receptors, most likely of the purine/pyrimidine subtype, enhance the ability of FGF2 to stimulate entry into a new cell cycle, as well as DNA replication, by an ERK dependent mechanism, whereas signaling by P2X receptors, possibly the P2X7 subtype, inhibits FGF2-induced mitogenesis in astrocytes. Interactions between P2Y, P2X and polypeptide growth factor signaling pathways may have important implications for CNS development as well as injury and repair. PMID- 18404518 TI - P2X(7) nucleotide receptors mediate caspase-8/9/3-dependent apoptosis in rat primary cortical neurons. AB - Apoptosis is a major cause of cell death in the nervous system. It plays a role in embryonic and early postnatal brain development and contributes to the pathology of neurodegenerative diseases. Here, we report that activation of the P2X(7) nucleotide receptor (P2X(7)R) in rat primary cortical neurons (rPCNs) causes biochemical (i.e., caspase activation) and morphological (i.e., nuclear condensation and DNA fragmentation) changes characteristic of apoptotic cell death. Caspase-3 activation and DNA fragmentation in rPCNs induced by the P2X(7)R agonist BzATP were inhibited by the P2X(7)R antagonist oxidized ATP (oATP) or by pre-treatment of cells with P2X(7)R antisense oligonucleotide indicating a direct involvement of the P2X(7)R in nucleotide-induced neuronal cell death. Moreover, Z DEVD-FMK, a specific and irreversible cell permeable inhibitor of caspase-3, prevented BzATP-induced apoptosis in rPCNs. In addition, a specific caspase-8 inhibitor, Ac-IETD-CHO, significantly attenuated BzATP-induced caspase-9 and caspase-3 activation, suggesting that P2X(7)R-mediated apoptosis in rPCNs occurs primarily through an intrinsic caspase-8/9/3 activation pathway. BzATP also induced the activation of C-jun N-terminal kinase 1 (JNK1) and extracellular signal-regulated kinases (ERK1/2) in rPCNs, and pharmacological inhibition of either JNK1 or ERK1/2 significantly reduced caspase activation by BzATP. Taken together, these data indicate that extracellular nucleotides mediate neuronal apoptosis through activation of P2X(7)Rs and their downstream signaling pathways involving JNK1, ERK and caspases 8/9/3. PMID- 18404516 TI - ATP release via anion channels. AB - ATP serves not only as an energy source for all cell types but as an 'extracellular messenger' for autocrine and paracrine signalling. It is released from the cell via several different purinergic signal efflux pathways. ATP and its Mg(2+) and/or H(+) salts exist in anionic forms at physiological pH and may exit cells via some anion channel if the pore physically permits this. In this review we survey experimental data providing evidence for and against the release of ATP through anion channels. CFTR has long been considered a probable pathway for ATP release in airway epithelium and other types of cells expressing this protein, although non-CFTR ATP currents have also been observed. Volume-sensitive outwardly rectifying (VSOR) chloride channels are found in virtually all cell types and can physically accommodate or even permeate ATP(4-) in certain experimental conditions. However, pharmacological studies are controversial and argue against the actual involvement of the VSOR channel in significant release of ATP. A large-conductance anion channel whose open probability exhibits a bell shaped voltage dependence is also ubiquitously expressed and represents a putative pathway for ATP release. This channel, called a maxi-anion channel, has a wide nanoscopic pore suitable for nucleotide transport and possesses an ATP binding site in the middle of the pore lumen to facilitate the passage of the nucleotide. The maxi-anion channel conducts ATP and displays a pharmacological profile similar to that of ATP release in response to osmotic, ischemic, hypoxic and salt stresses. The relation of some other channels and transporters to the regulated release of ATP is also discussed. PMID- 18404519 TI - A capillary electrophoresis method for the characterization of ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) and the analysis of inhibitors by in capillary enzymatic microreaction. AB - A capillary electrophoresis (CE) method for the characterization of recombinant NTPDases 1, 2, and 3, and for assaying NTPDase inhibitors has been developed performing the enzymatic reaction within the capillary. After hydrodynamic injection of plugs of substrate solution with or without inhibitor in reaction buffer, followed by a suspension of an enzyme-containing membrane preparation, and subsequent injection of another plug of substrate solution with or without inhibitor, the reaction took place close to the capillary inlet. After 5 min, the electrophoretic separation of the reaction products was initiated by applying a constant current of -60 muA. The method employing a polyacrylamide-coated capillary and reverse polarity mode provided baseline resolution of substrates and products within a short separation time of less than 7 min. A 50 mM phosphate buffer (pH 6.5) was used for the separations and the products were detected by their UV absorbance at 210 nm. The Michaelis-Menten constants (K (m)) for the recombinant rat NTPDases 1, 2, and 3 obtained with this method were consistent with previously reported data. The inhibition studies revealed pronounced differences in the potency of reactive blue 2, pyridoxalphosphate-6-azophenyl 2',4'-disulfonic acid (PPADS), suramin, and N (6)-diethyl-beta,gamma dibromomethylene-ATP (ARL67156) towards the NTPDase isoforms. Notably, ARL67156 does not inhibit all NTPDases, having only a minor inhibitory effect on NTPDase2. Dipyridamole is not an inhibitor of the NTPDase isoforms investigated. The new method is fast and accurate, it requires only tiny amounts of material (nanoliter scale), no sample pretreatment and can be fully automated; thus it is clearly superior to the current standard methods. PMID- 18404520 TI - The priming effect of extracellular UTP on human neutrophils: Role of calcium released from thapsigargin-sensitive intracellular stores. AB - P2Y(2) receptors, which are equally responsive to ATP and UTP, can trigger intracellular signaling events, such as intracellular calcium mobilization and mitogen-activated protein (MAP) kinase phosphorylation in polymorphonuclear leukocytes (PMN). Moreover, extracellular nucleotides have been shown to prime chemoattractant-induced superoxide production. The aim of our study was to investigate the mechanism responsible for the priming effect of extracellular nucleotides on reactive oxygen species (ROS) production induced in human neutrophils by two different chemoattractants: formyl-methionyl-leucyl phenylalanine (fMLP) and interleukin-8 (IL-8). Nucleotide-induced priming of ROS production was concentration- and time-dependent. When UTP was added to neutrophil suspensions prior to chemoattractant, the increase of the response reached the maximum at 1 min of pre-incubation with the nucleotide. UTP potentiated the phosphorylation of p44/42 and p38 MAP kinases induced by chemoattractants, however the P2 receptor-mediated potentiation of ROS production was still detectable in the presence of a SB203580 or U0126, supporting the view that MAP kinases do not play a major role in regulating the nucleotide-induced effect. In the presence of thapsigargin, an inhibitor of the ubiquitous sarco endoplasmic reticulum Ca(2+)-ATPases in mammalian cells, the effect of fMLP was not affected, but UTP-induced priming was abolished, suggesting that the release of calcium from thapsigargin-sensitive intracellular stores is essential for nucleotide-induced priming in human neutrophils. PMID- 18404521 TI - Estradiol inhibits the effects of extracellular ATP in human sperm by a non genomic mechanism of action. AB - Steroid hormones, beside their classical genomic mechanism of action, exert rapid, non genomic effects in different cell types. These effects are mediated by still poorly characterized plasma membrane receptors that appear to be distinct from the classic intracellular receptors. In the present study we evaluated the non genomic effects of estradiol (17betaE(2)) in human sperm and its effects on sperm stimulation by extracellular ATP, a potent activator of sperm acrosome reaction. In human sperm 17betaE(2) induced a rapid increase of intracellular calcium (Ca(2+)) concentrations dependent on an influx of Ca(2+) from the extracellular medium. The monitoring of the plasma membrane potential variations induced by 17betaE(2) showed that this steroid induces a rapid plasma membrane hyperpolarization that was dependent on the presence of Ca(2+) in the extracellular medium since it was absent in Ca(2+) free-medium. When sperm were pre-incubated in the presence of the K(+) channel inhibitor tetra-ethylammonium, the 17betaE(2) induced plasma membrane hyperpolarization was blunted suggesting the involvement of K(+) channels in the hyperpolarizing effects of 17betaE(2). Extracellular ATP induced a rapid plasma membrane depolarization followed by acrosome reaction. Sperm pre-incubation with 17betaE(2) inhibited the effects of extracellular ATP on sperm plasma membrane potential variations and acrosome reaction. The effects of 17betaE(2) were specific since its inactive steroisomer 17alphaE(2) was inactive. Furthermore the effects of 17betaE(2) were not inhibited by tamoxifen, an antagonist of the classic 17betaE(2) intracellular receptor. PMID- 18404522 TI - Molecular mechanisms of extracellular adenine nucleotides-mediated inhibition of human Cd4(+) T lymphocytes activation. AB - We have previously reported that ATPgammaS, a slowly hydrolyzed analog of ATP, inhibits the activation of human CD4(+) T lymphocytes by anti-CD3 and anti-CD28 mAb. In this report we have partially characterized the signaling mechanisms involved in this immunosuppressive effect. ATPgammaS had no inhibitory effect on CD4(+) T-cell activation induced by PMA and anti-CD28, indicating that it acts proximally to the TCR. It had no effect on the calcium rise induced by CD3/CD28 stimulation, but inhibited the phosphorylation of three kinases, ERK2, p38 MAPK and PKB, that play a key role in the activation of T cells. The receptor involved in these actions remains unidentified. PMID- 18404523 TI - ADPbetaS evokes microglia activation in the rabbit retina in vivo. AB - To investigate whether stimulation of purinergic P2Y(1) receptors modulates the activation of microglial and Muller glial cells in the rabbit retina in vivo, adenosine 5'-O-(2-thiodiphosphate) (ADPbetaS; 2 mM in 100 mul saline), a non hydrolyzable ADP analogue, was intravitreadly applied into control eyes or onto retinas that were experimentally detached from the pigment epithelium. Both retinal detachment and application of ADPssS onto control retinas induced phenotype alterations of the microglial cells (decrease of soma size, retraction of cell processes) and had no influence on microglial cell density. ADPssS application onto detached retinas accelerated the process retraction and resulted in a strongly decreased density of microglial cells. The effects of ADPssS on microglia density and phenotype in detached retinas were partially reversed by co application of the selective inhibitor of P2Y(1) receptors, MRS-2317 (3 mM in 100 mul saline). ADPssS apparently did not influence Muller cell gliosis, as determined by electrophysiological and calcium imaging records. It is concluded that rabbit retinal microglial cells express functional P2Y(1) receptors in vivo, and that activation of these receptors stimulates phenotype alterations that are characteristical for microglia activation. PMID- 18404525 TI - Contents of volume 1 (2005). PMID- 18404524 TI - LY 294002 inhibits adenosine receptor activation by a mechanism independent of effects on PI-3 kinase or casein kinase II. AB - Adenosine reduces both evoked and spontaneous calcium-dependent acetylcholine (ACh) release through a mechanism downstream of calcium entry at amphibian motor nerve endings (Silinsky EM. J Physiol 1984; 346: 243-56). LY 294002 (2-(4 morpholinyl)-8-phenyl-4H-1-benzopyran-4-one), an inhibitor of both phosphoinositide-3 kinase (PI-3 kinase) and casein kinase II, has been reported to increase spontaneous ACh release reflected in miniature endplate potential (MEPP) frequencies independently of intraterminal calcium at the frog neuromuscular junction (Rizzoli SO, Betz WJ. J Neurosci 2002; 22: 10680-9). It has been suggested that the increase in MEPP frequency caused by LY 294002, is mediated through an action on synaptotagmins, vesicle associated calcium sensors believed to trigger synaptic vesicle exocytosis. We thus examined the effects of adenosine on MEPP frequencies and evoked ACh release reflected as endplate potentials (EPPs) in order to determine if the presumed calcium-independent ACh release is affected by adenosine. We also wanted to determine if PI-3 kinase or casein kinase II is involved in mediating or modulating the inhibitory effects of adenosine. To these ends, we examined the effects of adenosine in the presence of LY 294002, wortmannin (a highly selective the PI-3 kinase inhibitor), or DRB (5,6 dichlorobenzimidazole riboside, an inhibitor of casein kinase II). LY 294002 reduced the sensitivity of both MEPP frequencies and the nerve-evoked calcium dependent EPPs to adenosine. The occlusive effects of LY 294002 on the actions of adenosine on MEPPs and EPPs were overcome by increasing adenosine concentration. Neither wortmannin nor DRB had any effect on the sensitivity of the EPPs to adenosine indicating that neither PI-3 kinase nor casein kinase II inhibition mediates the reduction in motor-nerve terminal sensitivity to adenosine produced by LY 294002. The results indicate a competitive relationship between LY 294002 and adenosine at A(1) receptors at the frog neuromuscular junction. This effect is independent of the previously described effects of LY 294002 on the exocytotic process, and is also independent of PI-3 kinase or casein kinase II. PMID- 18404527 TI - The contradictory effects of nitric oxide in caerulein-induced acute pancreatitis in rats. AB - This study was planned to observe the effects of nitric oxide synthesis on the antioxidative defense enzymes and pancreatic tissue histology in caerulein induced acute pancreatitis. Acute pancreatitis was induced by intraperitoneal injections of 50 microg/kg caerulein, L-arginine used for NO induction and N(omega)-nitro-L-arginine methyl ester (L-NAME) used for NO inhibition. In the caerulein group acinar cell degeneration, interstitial inflammation, oedema and haemorrhage were detected. Pancreatic damage scores were decreased with both NO induction and inhibition (p<0.05). MDA, GSH-Px, CAT, GSH and SOD activities were significantly changed in the caerulein group and indicated increased oxidative stress. Both NO induction and inhibition decreased this oxidative stress. It is concluded that both nitric oxide induction and inhibition ameliorated caerulein induced acute pancreatitis. The findings indicate that a certain amount of NO production has beneficial effects in experimental acute pancreatitis, but uncontrolled over-production of NO may be detrimental. PMID- 18404528 TI - Differential roles for Nrf2 and AP-1 in upregulation of HO-1 expression by arsenite in murine embryonic fibroblasts. AB - Heme oxygenase-1 (HO-1) is markedly upregulated by sodium arsenite and previous studies implicated the transcriptional enhancers Nrf2 and AP-1 in arsenite induced ho-1 gene expression in murine cells. To further evaluate the role of Nrf2 and its signalling pathway in the induction of HO-1 in response to low levels of arsenite, this paper studied wild-type and Nrf2-deficient murine embryonic fibroblasts. It was found that Nrf2 plays a crucial role in the early activation of ho-1 transcription and that increased Nrf2 levels returned to basal levels within 24 h. In Nrf2(-/-) cells, HO-1 gene activation increased gradually and HO-1 protein levels were approximately half of those attained in Nrf2(+/+) cells. The tyrosine kinase inhibitor genistein and JNK inhibitor SP600125 significantly attenuated arsenite induced increases in ho-1 mRNA levels in Nrf2 deficient cells but had negligible effects on Nrf2 activation, suggesting tyrosine kinase/JNK/c-Jun plays a key role in the HO-1 upregulation via AP-1. PMID- 18404529 TI - Effect of anaesthesia-induced alterations in haemodynamics on in vivo kinetics of nitroxyl probes in electron spin resonance spectroscopy. AB - Although the advent of in vivo electron spin resonance (ESR) spectroscopy has allowed analysis of the redox status of living animals, whether the haemodynamic condition affects the signal decay rate remains unknown. Three kinds of haemodynamic conditions were generated by changing the anaesthetic dosage in mice. Haemodynamics was analysed (n=6 each) and in vivo ESR was performed to measure the signal decay rates of three nitroxyl spin probes (carbamoyl-, carboxy and methoxycarbonyl-PROXYL) at the chest and head regions (n=6 for each condition and probe). Haemodynamic analysis revealed negative inotropic and chronotropic effects on the cardiovascular system depending on the depth of anaesthesia. Although signal decay rates differed among three probes, they were not affected by heart rate alteration. In this study we report the haemodynamics independent signal decay rate of nitoxyl probes. PMID- 18404530 TI - Lymphocyte antioxidant response and H2O2 production after a swimming session: gender differences. AB - This study evaluated the gender differences in response to intense exercise on lymphocyte hydrogen peroxide production, nitric oxide handling and mitochondrial superoxide dismutase (MnSOD) activity and gene expression. Fifteen males and nine females participated voluntarily in the study and performed a swimming session at 75-80% of the maximal capacity. In basal conditions females presented higher lymphocyte MnSOD activity compared to males (p<0.05). Exercise increased MnSOD activity in males (p<0.05) reaching similar values to females. MnSOD gene expression was also increased in males after exercise (p<0.05) but not in females. Nitrite concentration and iNOS gene expression significantly increased only in males after swimming (p<0.01). The exercise decreased UCP-3 gene expression in both genders (p<0.05). Lymphocyte H(2)O(2) production significantly increased in males after exercise in non-stimulated and in PMA-stimulated cells (p<0.01). In conclusion, females seem to be more protected against oxidative stress induced by a swimming session. Hydrogen peroxide is mainly produced in males and this subsequently leads to increases in MnSOD gene expression and activity. PMID- 18404531 TI - The protective effect of hypotaurine and cysteine sulphinic acid on peroxynitrite mediated oxidative reactions. AB - The protective activity of hypotaurine (HTAU) and cysteine sulphinic acid (CSA) on peroxynitrite-mediated oxidative damage has been assessed by monitoring different target molecules, i.e. tyrosine, dihydrorhodamine-123 (DHR) and glutathione (GSH). The inhibition of tyrosine oxidation exerted by HTAU and CSA both in the presence and the absence of bicarbonate can be ascribed to their ability to scavenge hydroxyl ((*)OH) and carbonate (CO(3)(*-)) radicals. HTAU and CSA also reduce tyrosyl radicals, suggesting that this repair function of sulphinates might operate as an additional inhibiting mechanism of tyrosine oxidation. In the peroxynitrite-dependent oxidation of DHR, the inhibitory effect of HTAU was lower than that of CSA. Moreover, while HTAU and CSA competitively inhibited the direct oxidation of GSH by peroxynitrite, HTAU was again poorly effective against the oxidation of GSH mediated by peroxynitrite-derived radicals. The possible involvement of secondary reactions, which could explain the difference in antioxidant activity of HTAU and CSA, is discussed. PMID- 18404532 TI - The differential NF-kB modulation by S-adenosyl-L-methionine, N-acetylcysteine and quercetin on the promotion stage of chemical hepatocarcinogenesis. AB - S-adenosylmethionine (SAM), N-acetylcysteine (NAC) and quercetin exhibit a chemoprotective effect. Likely this effect is mediated by counteracting, oxidative stress and NF-kB activation. To test this hypothesis F344 rats were subjected to hepatocarcinogenesis with or without antioxidants. NAC decreased foci in number and area, SAM and quercetin decreased area. Lipid-peroxidation was decreased by antioxidants, but only SAM increased glutathione. SAM, in its regulation from IKK downwards, abolished the NF-kB activation. NAC decreased IKK and IkB-a phosphorylation, and Rel-A/p65 and NF-kB binding, though the last two were affected with less intensity compared to the NF-kB inhibitor. Quercetin decreased Rel-A/p65, without modifying upstream signalling. Although all antioxidants inhibited oxidative stress as shown by reduction of lipid peroxidation, not all exerted the same effect on NF-kB signalling pathway and only SAM increased GSH. The mechanisms exerted by SAM in the reduction of foci makes this compound a potential liver cancer therapeutic agent. PMID- 18404533 TI - Antiserum detection of reactive carbonyl species-modified DNA in human colonocytes. AB - Polyunsaturated fats have been linked to occurrences of sporadic colon cancer. One possible cause may be degradation of polyunsaturated fats during cooking, resulting in multiple reactive carbonyl species (RCS) that can damage nuclear DNA and proteins, particularly in rapidly dividing colon crypt cells. This study describes a novel antiserum against RCS-modified DNA, with apparent order of reactivity to DNA modified with 4-hydroxy-trans-2-nonenal > glyoxal > acrolein > crotonaldehyde > malondialdehyde; some reactivity was also observed against conjugated Schiff base-type structures. Anti-(RCS-DNA) antiserum was successfully utilised to demonstrate formation of RCS-DNA in a human colon cell model, exposed to RCS insult derived from endogenous and exogenous lipid peroxidation sources. Further utilisation of the antiserum for immunohistochemical analysis confirmed RCS-modified DNA in crypt areas of 'normal' colon tissue. These results fully support a potential role for dietary lipid peroxidation products in the development of sporadic colon cancer. PMID- 18404534 TI - The role of oxidative stress in acrolein-induced DNA damage in HepG2 cells. AB - This study evaluated the role of oxidative stress in acrolein-induced DNA damage, using HepG2 cells. Using the standard single cell gel electrophoresis (SCGE) assay, a significant dose-dependent increment in DNA migration was detected at lower concentrations of acrolein; but at the higher tested concentrations, a reduction in the migration was observed. Post-incubation with proteinase K significantly increased DNA migration in cells exposed to higher concentrations of acrolein. These results indicated that acrolein caused DNA strand breaks and DNA-protein crosslinks (DPC). To elucidate the oxidatively generated DNA damage mechanism, the 2,7-dichlorofluorescein diacetate (DCFH-DA) and o-phthalaldehyde (OPT) were used to monitor the levels of reactive oxygen species (ROS) and glutathione (GSH), respectively. The present study showed that acrolein induced the increased levels of ROS and depletion of GSH in HepG2 cells. Moreover, acrolein significantly caused 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) formation in HepG2 cells. These results demonstrate that the DNA damage induced by acrolein in HepG2 cells is related to the oxidative stress. PMID- 18404535 TI - Protection of chlorophyllin against oxidative damage by inducing HO-1 and NQO1 expression mediated by PI3K/Akt and Nrf2. AB - Green vegetables are thought to have a chemoprotective effect on the basis of epidemiologic evidence. This study investigated whether chlorophyllin (CHL) could induce antioxidant enzymes and confer protection against oxidative damage. The results showed that CHL could induce HO-1 and NQO1 expression in human umbilical vein endothelial cell (HUVEC) in a time- and dose-dependent manner and protect them against hydrogen peroxide caused oxidative damage. The induction of HO-1 and NQO1 by CHL was accompanied with the accumulation of transcription factor Nrf2 in nucleus and the activation of PI3K/Akt signalling pathway. Additionally, the specific inhibitor of PI3K/Akt could obviously decrease not only the induced expression of HO-1 and NQO1 but also the antioxidant effect of CHL. In conclusion, this study proved that CHL exerts antioxidant effect by inducing HO-1 and NQO1 expression mediated by PI3K/Akt and Nrf2. One thinks CHL may have promise to be prophylactic pharmaceuticals without adverse effects. PMID- 18404536 TI - Antioxidant and free radical scavenging properties of N-acetylcysteine amide (NACA) and comparison with N-acetylcysteine (NAC). AB - The antioxidant potential of N-acetylcysteine amide (NACA), also known as AD4, was assessed by employing different in vitro assays. These included reducing power, free radical scavenging capacities, peroxidation inhibiting activity through linoleic acid emulsion system and metal chelating capacity, as compared to NAC and three widely used antioxidants, alpha-tocopherol, ascorbic acid and butylated hydroxytoluene (BHT). Of the antioxidant properties that were investigated, NACA was shown to possess higher 2,2-diphenyl-1-picryl-hydrazyl hydrate (DPPH) radical scavenging ability and reducing power than NAC, at all the concentrations, whereas the scavenging ability of H(2)O(2) differed with concentration. While NACA had greater H(2)O(2) scavenging capacity at the highest concentration, NAC was better than NACA at lower concentrations. NAC and NACA had a 60% and 55% higher ability to prevent beta-carotene bleaching, respectively, as compared to control. The chelating activity of NACA was more than 50% that of the metal chelating capacity of EDTA and four and nine times that of BHT and alpha tocopherol, respectively. When compared to NACA and NAC; alpha-tocopherol had higher DPPH scavenging abilities and BHT and alpha-tocopherol had better beta carotene bleaching power. These findings provide evidence that the novel antioxidant, NACA, has indeed enhanced the antioxidant properties of NAC. PMID- 18404537 TI - In vitro electron paramagnetic resonance characterization of free radicals: relevance to exercise-induced lipid peroxidation and implications of ascorbate prophylaxis. AB - This study tested the hypothesis that exercise-induced oxidative stress is caused by free radical-mediated damage to polyunsaturated fatty acids (PUFA) which can be prevented following ascorbate prophylaxis. Hyperfine coupling constants (HCC) of alpha-phenyl-tert-butylnitrone (PBN)-adducts were measured via room temperature electron paramagnetic resonance (EPR) spectroscopy in the venous blood of 12 subjects at rest and following maximal exercise during a randomized double-blind placebo-controlled trial and compared to those observed following room-air incubation (2 h at 37 degrees C) of L-alpha-phosphatidycholine, linoleic acid, alpha-linolenic acid and arachidonic acid. All adducts exhibited similar HCC [a(N) 13.6 Gauss (G) and a beta(H) 1.8 G] with the exception of L-alpha phosphatidycholine [a(N1)=13.4 G, a beta(H1)=1.6 G (37%) and a(N2)=14.9 G, a beta(H2)=0.3 G (63%)] consistent with the trapping of lipid-derived alkoxyl and oleate radicals, respectively. Ascorbate pre-treatment ablated radical formation in both systems. These findings identify circulating PUFA as a potential source of secondary radicals that are capable of initiating oxidative stress in the exercising human. PMID- 18404539 TI - Change in histone H3 phosphorylation, MAP kinase p38, SIR 2 and p53 expression by resveratrol in preventing streptozotocin induced type I diabetic nephropathy. AB - Resveratrol has been reported to have a wide variety of biological effects. However, little is known regarding its role on phosphorylation of histone H3, MAP kinase p38, SIR2 and p53 in type I diabetic nephropathy (DN). Hence, the present study was undertaken to examine changes in the above said parameters by resveratrol treatment. Male Sprague-Dawley rats were rendered diabetic using a single dose of streptozotocin (55 mg/kg, i.p.). DN was assessed by measurements of blood urea nitrogen and creatinine levels. Phosphorylation of histone H3, SIR2, p53 and MAP kinase p38 expression were examined by western blotting. This study reports that treatment of resveratrol prevents the decrease in the expression of SIR2 in diabetic kidney. It also prevents increase in p38, p53 expression and dephosphorylation of histone H3 in diabetic kidney. This is the first report which suggests that protection against development of diabetic nephropathy by resveratrol treatment involves change in phosphorylation of histone H3, expression of Sir-2, p53 and p38 in diabetic kidney. PMID- 18404540 TI - Effect of nonsteroidal anti-inflammatory drugs and the procarcinogen 1,2 dimethylhydrazine on the antioxidant defense system. AB - The present study was designed to evaluate the effects of three nonsteroidal anti inflammatory drugs (NSAIDs) with varying cycloxygenase selectivities on the small intestinal antioxidant enzyme status and surface characteristics during 1,2 dimethylhydrazine (DMH) administration. Male Sprague-Dawley rats were divided into five different groups: Group 1 (control, vehicle treated); group 2 (DMH treated, 30 mg/kg body weight/week, subcutaneously); group 3 (DMH + aspirin 60 mg/kg body weight); group 4 (DMH + celecoxib 6 mg/kg body weight); group 5 (DMH + etoricoxib 0.64 mg/kg body weight). Postmitochondrial fraction were isolated from the intestinal segments and different oxidative parameters and other parameters studied, such as the lipid peroxides, reduced and total glutathione, superoxide dismutase, catalase, glutathione reductase, glutathione S-transferase, nitric oxide, citrulline, and nucleic acids. At the end of 6 weeks of treatment, the results indicated a significant alteration in the antioxidative defense status of the intestine in the presence of the procarcinogen DMH, which was restored with the administration of NSAIDs. The study, therefore, suggests a possible mechanism for the chemopreventive effects of NSAIDs against the experimental intestinal cancer in rats. PMID- 18404538 TI - Severe vitamin E deficiency modulates airway allergic inflammatory responses in the murine asthma model. AB - Allergic asthma is a complex immunologically mediated disease associated with increased oxidative stress and altered antioxidant defenses. It was hypothesized that alpha-tocopherol (alpha-T) decreases oxidative stress and therefore its absence may influence allergic inflammatory process, a pathobiology known to be accompanied by oxidative stress. Therefore, selected parameters of allergic asthma sensitization and inflammation were evaluated following ovalbumin sensitization and re-challenge of alpha-T transfer protein (TTP) knock-out mice (TTP(-/-)) that have greatly reduced lung alpha-T levels (e.g.<5%) compared to their litter mate controls (TTP(+/+)). Results showed that severe alpha-T deficiency result in a blunted lung expression of IL-5 mRNA and IL-5 protein and plasma IgE levels compared with TTP(+/+) mice following immune sensitization and rechallenge, although lung lavage eosinophil levels were comparable in both genomic strains. It is concluded that the initial stimulation of immune responses by the TTP(-/-) mice were generally blunted compared to the TTP(+/+) mice, thus diminishing some aspects of subsequent allergic inflammatory processes. PMID- 18404541 TI - Inhibitory effect of tributyltin on expression of steroidogenic enzymes in mouse testis. AB - Tributyltin (TBT) is known to disrupt the development of reproductive organs, thereby reducing fertility. The aim of this study was to evaluate the acute toxicity of TBT on the testicular development and steroid hormone production. Immature (3-week-old) male mice were given a single administration of 25, 50, or 100 mg/kg of TBT by oral gavage. Lumen formation in seminiferous tubule was remarkably delayed, and the number of apoptotic germ cells found inside the tubules was increased in the TBT-exposed animals, whereas no apoptotic signal was observed in interstitial Leydig cells. Reduced serum testosterone concentration and down-regulated expressions of the mRNAs for cholesterol side-chain cleavage enzyme (P450scc), 17alpha -hydroxylase/C(17-20) lyase (P450(17alpha)), 3beta hydroxysteroid-dehydrogenase (3beta -HSD), and 17beta -hydroxysteroid dehydrogenase (17beta -HSD) were also observed after TBT exposure. Altogether, these findings demonstrate that exposure to TBT is associated with induced apoptosis of testicular germ cells and inhibition of steroidogenesis by reduction in the expression of steroidogenic enzymes in interstitial Leydig cells. These adverse effects of TBT would cause serious defects in testicular development and function. PMID- 18404542 TI - Evaluation of the developmental toxicity of linalool in rats. AB - The developmental toxicity of linalool, a widely used fragrance ingredient, was evaluated in presumed pregnant Sprague-Dawley rats (25/group). Oral dosages of 0, 250, 500, or 1000 mg/kg/day linalool were administered by gavage on gestational days 7 to 17. The presence of spermatozoa and/or a copulatory plug in situ was designated as gestational day 0. Rats were observed for viability, clinical signs, body weights, and feed consumption. Caesarean sectioning and necropsy occurred on gestational day 21. Uteri were examined for number and distribution of implantations, live and dead fetuses, and early and late resorptions. Numbers of corpora lutea were also recorded. Fetuses were weighed and examined for gender, gross external changes, and soft tissue or skeletal alterations. There were no maternal deaths, clinical signs, or gross lesions that were considered related to linalool. During the dosage period, mean relative feed consumption was significantly reduced by 7% and mean body weight gains were reduced by 11% at 1000 mg/kg/day. During the postdosage period, feed consumption values at 1000 mg/kg/day were significantly higher than vehicle control values, which corresponded to the increase in body weight gains during this period. Caesarean section and litter parameters, as well as fetal alterations, were not affected by linalool at any of the three dosages tested. On the basis of these data, the maternal no observed adverse effect level (NOAEL) of linalool is 500 mg/kg/day, whereas the developmental NOAEL is > or = 1000 mg/kg/day. It is concluded that linalool is not a developmental toxicant in rats at maternal doses of up to 1000 mg/kg/day. PMID- 18404543 TI - Subchronic oral toxicity of ubiquinol in rats and dogs. AB - Ubiquinol is the two-electron reduction product of ubiquinone (coenzyme Q(10) or CoQ(10)) and functions as an antioxidant in both mitochondria and lipid membranes. In humans and most mammals, including dogs, the predominant form of coenzyme Q is coenzyme Q(10), whereas the primary form in rodents is coenzyme Q(9) (CoQ(9)). Therefore, the subchronic toxicity of ubiquinol was evaluated and compared in Sprague-Dawley rats and beagle dogs. In the initial rat study, males and females were given ubiquinol at doses of 0, 300, 600, or 1200 mg/kg or ubiquinone at 1200 mg/kg by gavage for 13 weeks. This was followed by the second study, where females were given with doses of 75, 150, 200, or 300 mg/kg/day in order to determine a no observed adverse effect level (NOAEL). In the dog study, the test material was administered to males and females at dose levels of 150, 300, and 600 mg/kg, and ubiquinone was included at 600 mg/kg. Clinical observations, mortality, body weights, food and water consumption, ophthalmoscopy, urinalysis, hematology, blood biochemistry, gross findings, organ weights, and histopathological findings were examined. In both species, determination of plasma and liver ubiquinol concentrations, measured as total coenzyme Q(10), were performed. There were no deaths or test article-related effects in body weight, food consumption, ophthalmology, urinalysis, or hematology in rats. Histopathological examinations revealed test article-related effects on the liver, spleen, and mesenteric lymph node in female rats but not in male rats. In the liver, fine vacuolation of hepatocytes was observed in the ubiquinol groups at 200 mg/kg and above. These changes were judged to be of no toxicological significance because they were not considered to induce cytotoxic changes. Microgranuloma and focal necrosis with accumulation of macrophages were observed in the ubiquinol groups at 300 mg/kg and above. These findings were accompanied by slight increases in blood chemistry enzymes (aspartate aminotransferase [AST], alanine aminotransferase [ALT], and lactate dehydrogenase [LDH]), which was suggestive of either potential hepatotoxicity or a normal physiological response to ubiguinol loading. Microgranuloma, and focal necrosis were judged to be only adverse effects induced by test article based on their incidence and pathological characteristics. These changes observed in liver were thought due to uptake of the administered ubiquinol by the liver as an adaptive response to xenobiotics, and the microgranulomas and focal necrosis were considered the results of excessive uptake of ubiquinol, which exceeded the capacity for adaptive response. Based on these findings the NOAEL in rats was conservatively estimated to be 600 mg/kg/day for males and 200 mg/kg/day for females. In dogs, there were no deaths or ubiquinol-related toxicity findings during the administration period. No test article-related effects were observed in body weight, food consumption, ophthalmology, electrocardiogram, urinalysis, hematology, or blood chemistry. Histopathological examination revealed no effects attributable to administration of ubiquinol or ubiquinone in any organs examined. Based on these findings, a NOAEL for ubiquinol in male and female dogs was estimated to be more than 600 mg/kg/day under the conditions of this study. PMID- 18404544 TI - Effects of vitamin E, vitamin C, and selenium on gastric fundus in cadmium toxicity in male rats. AB - Cadmium (Cd) is a highly toxic metal. It has an indirect role in the generation of various free radicals. Antioxidants such as vitamin E, vitamin C, and selenium are important for preventing the damage caused by reactive oxygen species. This study was undertaken to examine the effect of acute cadmium and/or antioxidants on serum lipid metabolism, tissue glutathione, and lipid peroxidation (LPO) levels, and ghrelin and metallothionein production in the gastric fundus mucosa of rats. Cd (2 mg/kg/day CdCl(2)) was administered to rats for 8 days, intraperitoneally. Vitamin E (250 mg /kg/day) + vitamin C (250 mg/kg/day) + sodium selenate (0.25 mg /kg/day) were administered to rats orally at the same time. The animals were treated by antioxidants 1 h prior to treatment with Cd every day. Gastric tissue homogenates were used for protein and glutathione and LPO levels. Phospholipid and total lipid levels were determined in serum. Gastric fundus sections examined for histopathological changes and by immunohistochemistry for expression of ghrelin and metallothionein. In the group treated with Cd, degenerative changes such as discontinuity in the surface epithelium were observed. The degenerative changes induced by Cd were decreased in the group given vitamin E + vitamin C + selenium. There was no significant change in ghrelin- and metallothionein-immunoreactive cells in fundus mucosa. Stomach glutathione levels insignificantly decreased in the Cd groups, but in the Cd group given antioxidant, stomach glutathione levels were significantly increased. Serum phospholipid and total lipid levels were significantly increased in the Cd groups. On the other hand, treatment with antioxidants reversed these effects. These results indicate that antioxidants partly prevent the toxicity of Cd in rat gastric fundus. PMID- 18404545 TI - What's so special about the developing immune system? AB - The evolution of the subdiscipline of developmental immunotoxicology (DIT) as it exists today has been shaped by significant regulatory pressures as well as key scientific advances. This review considers the role played by legislation to protect children's health, and on the emergence of immunotoxcity and developmental immunotoxicity guidelines, as well as providing some context to the need for special attention on DIT by considering the evidence that the developing immune system may have unique susceptibilities when compared to the adult immune system. Understanding the full extent of this potential has been complicated by a paucity of data detailing the development of the immune system during critical life stages as well as by the complexities of comparisons across species. Notably, there are differences between humans and nonhuman species used in toxicity testing that include specific differences relative to the timing of the development of the immune system as well as more general anatomic differences, and these differences must be factored into the interpretation of DIT studies. Likewise, understanding how the timing of the immune development impacts on various immune parameters is critical to the design of DIT studies, parameters most extensively characterized to date in young adult animals. Other factors important to DIT, which are considered in this review, are the recognition that effects other than suppression (e.g., allergy and autoimmunity) are important; the need to improve our understanding of how to assess the potential for DIT in humans; and the role that pathology has played in DIT studies in test animals. The latter point receives special emphasis in this review because pathology evaluations have been a major component of standard nonclinical toxicology studies, and could serve an important role in studies to evaluate DIT. This possibility is very consistent with recommendations to incorporate a DIT evaluation into standard developmental and reproductive toxicology (DART) protocols. The overall objective of this review is to provide a 'snapshot' of the current state-of-the-science of DIT. Despite significant progress, DIT is still evolving and it is our hope that this review will advance the science. PMID- 18404547 TI - Exophiala spinifera and its allies: diagnostics from morphology to DNA barcoding. AB - Diagnostic features of morphology, physiology, serology and genetics of species belonging to the Exophiala spinifera clade (including 11 species: Exophiala oligosperma, E. spinifera, E. xenobiotica, E. jeanselmei, E. exophialae, E. nishimurae, E. bergeri, E. nigra, Rhinocladiella similis, Ramichloridium basitonum and Phaeoannellomyces elegans), comprising a large number of human associated Exophiala species, are summarized. Several species have closely similar morphological characters and physiological profiles. Taxonomy is therefore primarily based on sequence diversity of the Internal Transcribed Spacer (ITS) region of ribosomal DNA (rDNA). Multilocus sequencing has shown that ITS is reliable for identification of the species in this clade, and is a therefore a good candidate for barcoding species of Exophiala. Species-specific fragments were searched in the ITS region of species in the Exophiala spinifera clade and can be used to design probes for diagnosis by hybridization. PMID- 18404548 TI - Investigation into the production of 2-Pentylfuran by Aspergillus fumigatus and other respiratory pathogens in vitro and human breath samples. AB - Our objective was to identify, for diagnostic purposes, potential volatile biomarkers of human microbial pathogens. We analysed the head space of cultures of medically important bacterial and fungal respiratory pathogens for 2 Pentylfuran (2PF) production through the use of Solid Phase Micro Extraction (SPME) and Gas Chromatography/Mass Spectroscopy (GC/MS). Our results confirm that 2PF is consistently produced by Aspergillus fumigatus, Fusarium spp., Aspergillus terreus, Aspergillus flavus and to a lesser extent by Aspergillus niger. 2 Pentylfuran was not detected from most of the bacterial strains except for Streptococcus pneumoniae. In human studies, four litre breath samples were collected from patients with cystic fibrosis (CF), with and without colonisation by A. fumigatus and other pathogens, as well as healthy volunteers. 2-Pentylfuran was detected in breath samples collected from 4/4 patients with CF and A. fumigatus colonization, 3/7 patients with CF and no microbiological evidence of A. fumigatus and 0/10 healthy control individuals. These results suggest that 2PF may be a biomarker for lung colonization/infection by fungal pathogens. To our knowledge, this is the first report describing the detection in breath samples of a volatile biomarker of a pathogen resident in the lungs. Breath analysis has the potential of being a non-invasive diagnostic method of detecting respiratory infection including invasive aspergillosis. PMID- 18404549 TI - Examining Trichophyton tonsurans genotype and biochemical phenotype as determinants of disease severity in tinea capitis. AB - Trichophyton tonsurans infections occur in various host populations, on various body sites and with varying degrees of inflammation. This investigation was undertaken to determine whether fungal factors could explain the degree of severity in clinical symptomatology among infected children. Otherwise healthy children (n=54) presenting with tinea capitis were enrolled in this study. A thorough history was performed, the extent and severity of infection graded and a fungal specimen collected from each child. Strain type was determined by genotyping for 11 sequence variations in the rDNA and ALP1 loci. Secreted protease activity was quantitated after 5 days of growth in aqueous medium. Forty participants were evaluable. Infection duration ranged from 1 day to 3 years and clinical severity score (CSS) from 4-19. Seventeen unique fungal genotypes were present. Keratinase, collagenase and elastase activity varied 32.7-fold, 64.9 fold and 303.3-fold, respectively. A significant association was observed between genotype and disease severity with the rDNA sequence variations accounting for over 50% of the variation observed in CSS (r2=0.539; P<0.001). Phylogenetic analyses appear to suggest that the ancestral strain types of T. tonsurans cause more severe disease. These observations are consistent with reports that recently diverge anthropophilies are associated with diminished inflammatory involvement. PMID- 18404550 TI - Workload and clinical significance of the isolation of zygomycetes in a tertiary general hospital. AB - Several reports of increases in invasive zygomycosis (IZ) at individual institutions across the USA and Europe have contributed to a generalized concept that IZ is an increasing problem and the overestimation of the clinical significance of the isolation of zygomycetes in microbiology departments. We assessed the workload and clinical significance of zygomycetes isolates recovered from clinical samples in our institution over a 19-year period (1988-2006). We retrospectively reviewed the charts of those patients from who isolates of zygomycetes were obtained and calculated the workload of its isolation, the incidence of IZ during this period and the positive predictive value (PPV) of a positive culture. Zygomycetes were recovered from 210 samples (176 patients), i.e., 0.086/1,000 clinical samples processed and 6.3/1,000 samples submitted for fungal isolation. Zygomycetes represented 0.6% of the total fungi recovered. The mean incidence of the disease was 1.2 cases/100,000 admissions (range 0-20). Only 16 of the samples which grew zygomycetes (7.6%) were from infected patients. The workload generated by zygomycetes in our institution and the PPV for IZ of their isolation in our laboratory were very low and the disease was not found to have significantly increased in recent years in our institution. Data from specific institutions cannot be generalized. PMID- 18404552 TI - Human neutrophils susceptibility to Paracoccidioides brasiliensis: an ultrastructural and cytochemical assay. AB - Paracoccidioidomycosis, caused by the dimorphic fungus Paracoccidioides brasiliensis, is the most prevalent systemic mycosis of Latin America, with Brazil accounting for 80% of the reported cases. The great number of neutrophils found in P. brasiliensis granulomas demonstrates the importance of polymorphonuclear neutrophils (PMN) cells during this mycotic infection. It has been found that neutrophils from healthy human donors can ingest and kill the fungus through a typical phagocytic process. The present work tests the phagocytic ability of neutrophils collected from patients that had had and were considered cured of paracoccidioidomycosis. Transmission electron microscopy and cytochemical studies indicate that patients' neutrophils eventually degenerate during phagocytosis of P. brasiliensis. Endogen peroxidase and NAD(P)H-oxidase are activated during the process showing that the respiratory burst and the neutrophil degranulation are triggered by the attachment of the yeast cells. Apparently these processes are not enough to kill P. brasiliensis. Although fungicidal activity can be determined by colony forming unit (CFU) counting, qualitative data suggest, as noted, that neutrophils from patients with treated paracoccidioidomycosis degenerate during the phagocytosis process. Hence, this work demonstrates the existence of a functional neutrophil deficiency against P. brasiliensis in susceptible individuals. The exact origin of this susceptibility is still to be determined in further studies. PMID- 18404551 TI - Characterization of caspofungin susceptibilities by broth and agar in Candida albicans clinical isolates with characterized mechanisms of azole resistance. AB - Caspofungin (CSP) susceptibilities of Candida albicans, as determined by broth microdilution methods, have not been found to be related to azole susceptibilities or resistance. In contrast, it has been observed that azole resistant clinical isolates that overexpress the efflux pump gene CDR2 are less susceptible to CSP when tested using an agar dilution method commonly employed with Saccharomyces cerevisiae. The goal of this study was to further understand the effects of azole resistance mechanisms on CSP susceptibility testing. A collection of 69 isolates exhibiting known mechanisms of azole resistance and resistance-associated phenotypes were analyzed by broth microdilution methods to determine standard minimum inhibitory concentrations (MICs) for CSP. The same isolates were then analyzed as to their MIC to CSP by Etest strips, an agar-based method that has been shown generally to be comparable to broth methods. The MICs found with both methods were not significantly different. However, a collection of strains overexpressing the efflux pump CDR2 did exhibit a spectrum of CSP susceptibilities when examined by agar dilution susceptibility tests, ranging from standard to reduced susceptibilities. This work demonstrated that a change in CSP susceptibility with CDR2 overexpressing cells in agar dilution studies is a variable phenotype and it is not the result of growth conditions (i.e., broth versus agar). PMID- 18404553 TI - Candida albicans metabolite affects the cytoskeleton and phagocytic activity of murine macrophages. AB - Candida albicans is the most common opportunistic fungal pathogen of humans, causing systemic disease in immunocompromised patients. Host resistance to C. albicans infections is mediated predominantly by neutrophils and monocytes/macrophages. We have previously shown that exposure of a human epithelial cell line (HEp2) to C. albicans or to a culture filtrate of C. albicans caused actin rearrangement in the HEp2 cells. Since shifting of actin from the filamentous to the globular form may be crucial to the activity of phagocytes, we assessed in the present study the effect of the C. albicans metabolite (lyophilized culture filtrate) on the cytoskeleton of murine peritoneal macrophages and on their phagocytic activity. Our results showed a significant decrease in phagocytosis of C. albicans, ranging from 53-63% and a 25% reduction for C. glabrata cells. Using confocal laser scanning microscopy an actin rearrangement in the macrophages could be demonstrated that may be associated with the decrease of phagocytosis. We also tested the effect of mannan and of the secreted aspartic proteinase (Sap) inhibitor--pepstatin, on the activity of the metabolite in order to define the putative component and found no influence. In conclusion, our data indicate that a C. albicans metabolite affects phagocytic activity of macrophages, probably by alterations in their cytoskeleton. PMID- 18404554 TI - Cholesterol dependent and Amphotericin B resistant isolates of a Candida glabrata strain from an Intensive Care Unit patient. AB - Here we report on two isolates of Candida glabrata recovered from urine samples collected from of an Intensive Care Unit patient. D1/D2 and ITS 1+2 rDNA sequence analysis confirmed its identification. The isolates were cholesterol dependent and resistant to Amphotericin B. PMID- 18404555 TI - Disseminated cryptococcosis with necrotizing fasciitis in an apparently immunocompetent host: a case report. AB - We present an unusual case of disseminated cryptococcosis with necrotizing fascitis in an immunocompetent host. The multi-system cryptococcal infection was proved using standard microbiological, histopathological and radiological investigations. After a combination of antifungal, antimicrobial and surgical therapy clinical cure was achieved. PMID- 18404556 TI - Disseminated fungal infection in a renal transplant recipient involving Macrophomina phaseolina and Scytalidium dimidiatum: case report and review of taxonomic changes among medically important members of the Botryosphaeriaceae. AB - We report the first case of human infection with the fungal plant pathogen Macrophomina phaseolina in a Sri Lankan-born Canadian man following a renal transplant in India. The patient subsequently succumbed to invasive infection with Scytalidium dimidiatum. Molecular sequence analysis confirmed the identification of both fungi and revealed that they are related species within the ascomycete family Botryosphaeriaceae. We review the rationale for the recent reclassification of S. dimidiatum as Neoscytalidium dimidiatum and of Nattrassia mangiferae (formerly considered a synanamorph of S. dimidiatum) as Neofusicoccum mangiferae. This and other recent cases illustrate the potential for plant pathogenic fungi to cause invasive human diseases which are refractory to antifungal therapy. PMID- 18404557 TI - A new benzofuranic acid from the leaves of Rhus alata. AB - From leaves of Rhus alata, one new benzofuranic acid named [(2E)-3-(4-hydroxy-5,7 dimethyl- benzo[3,4-b] furan-6-yloxy)-prop-2-enoic acid has been isolated together with eight known compounds: dimethyl ester of terephthalic acid, beta amyrin, friedelin, lupeol, beta-sitosterol, oleanolic acid, taraxerone and ethyl gallate. Structural elucidations were done on the basis of chemical and physical data (IR, UV, 1H-NMR, 13C-NMR and MS spectra). PMID- 18404558 TI - Synthesis and structure-activity relationship study of monotesone-A, an antifungal component of Monotes engleri. AB - The synthesis of (+/-)-monotesone-A, an antifungal component of Monotes engleri, as well as its four structural analogues were accomplished starting from the corresponding MOM-protected phloracetophenone and benzaldehyde derivatives. Antifungal activities of the five flavanone derivatives were studied against Candida albicans (14053 and ATCC 10231), Candida inconspicua, Candida dubliniensis and Candida krusei. PMID- 18404559 TI - Composition and antimicrobial activity of the essential oil of Acronychia pedunculata (L.) Miq. from Vietnam. AB - The chemical composition and the antimicrobial activity of the essential oil isolated from aerial parts of Acronychia pedunculata (L.) Miq. from Vietnam are reported. Analysis was carried out by GC (RI), GC-MS and 13C NMR. Thirty-four compounds were identified, accounting for 92.8% of the oil. The major constituents were alpha-pinene (57.4%) and (E)-beta-caryophyllene (13.6%). The essential oil of A. pedunculata was shown to possess a broad spectrum of antimicrobial activity against various bacteria, particularly Salmonella enterica and Staphylococcus epidermidis. PMID- 18404560 TI - Computational studies of Fe(III) binding to bryostatins, bryostatin analogs, siderophores and marine natural products: arguments for ferric complexes in medicinal applications. AB - In this computational study, geometric factors are calculated by applying semi empirical methods (PM3) that support experimental evidence from this lab where bryostatins can bind trivalent iron with six Fe-O bonds forming an octahedral geometry. The geometric factors are calculated for all 20 structures (Fe3+ bound to bryostatin 1-20) as a neutral, monovalent, and divalent species. The average Fe-O bond distances and bond angles are compared to those of known marine and terrestrial siderophores. From these two data sets, we then examined other known marine natural products (MNPs) that can form a hexavalent complex with six Fe-O bonds and draw conclusions about their potential biological role as marine siderophores. This computational data indicates that Fe(III) strongly bonds to a host of MNPs, increasing their water solubility, contracting their structure, hence allowing transport through cell membranes more readily, and in some cases, stabilizing ester bonds that are susceptible to hydrolysis. It is argued that administering medicinally bryostatin, its analogs or other MNPs as a ferric complex, holds some fundamental chemical advantages compared to its administration as a neutral uncomplexed species. PMID- 18404561 TI - Isolation of beta-glucan from the cell wall of Saccharomyces cerevisiae. AB - Beta-glucan, one of the major cell wall components of Saccharomyces cerevisiae (S. cerevisiae), has been found to enhance immune functions. At present study, we developed an optimal procedure to extract and purify beta-glucan. At first, yeast cells were grown in sabouraud dextrose agar and then cultured in yeast extract peptone-glucose (YPG) broth. After incubation, cells were harvested, washed and disrupted by means of sonication method. The obtained cell walls were used to prepare alkali-soluble beta-glucan (glucan-S1). In this regard, 2% sodium hydroxide (NaOH) and 3% acetic acid were used in alkaline-acid extraction, respectively. This preparation contained 2.4% protein. In the next step, DEAE sephacel chromatography was used to remove remaining proteins (glucan-S2). Subsequently this preparation was applied into concanavalin-A sepharose column to remove manann. Finally, beta-glucan free of mannoprotein complexes was prepared (glucan-S3). PMID- 18404562 TI - An experimental study of the effects of Matricaria chamomilla extract on cutaneous burn wound healing in albino rats. AB - Previous studies conducted on the anti-inflammatory, antimicrobial and antioxidant effects of Matricaria chamomilla (chamomile) extract led us to study the effect of topical chamomile extract on burn wound healing in albino rats. Thirty male albino rats (250-300 g) were randomly divided into three groups, as control, vehicle, and treatment. Second-degree burning was induced in 20% of whole surface area of animal body by placing the back of animal into boiling water for 8s. Animals of control group received no treatment. Animals of vehicle and treatment groups were treated topically by olive oil and extract dissolved in olive oil twice a day respectively from the first day of burn induction to complete wound healing. The percentage of wound healing was calculated weekly. The results showed that there was significant difference (p < 0.05) between vehicle and treatment groups. So we concluded that the chamomile extract in the form of rubbing oil had a good potential for acceleration of burn wound healing in rats. PMID- 18404563 TI - Phytotherapeutic inhibition of supragingival dental plaque. AB - Antimicrobial activities and biofilm-formation preventive properties of Mentha piperita and Cuminum cyminum essential oils and chlorhexidine were assessed against Streptococcus mutans and Streptococcus pyogenes. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) analysis led to the identification of 26 and 32 compounds in the essential oils of M. piperita and C. cyminum, respectively. Minimal bactericidal concentrations (MBC) of the oils and chlorhexidine and microbial decimal reduction time (D value) were determined. Antibacterial and in vivo biofilm preventive efficacies of all the concentrations of M. piperita oil were significantly (p<0.001) higher. The biofilm inhibitory properties in planktonic cultures were in M. piperita > chlorhexidine > C. cyminum order. In vivo experiments conducted on male and female volunteers who brushed with essential oil blended toothpastes indicated that lower concentrations of the oils, in particular the M. piperita oil, were significantly higher (p<0.001) and effective during the course of the study as compared to chlorhexidine. In conclusion, there may be a potential role for essential oils in the development of novel anticaries treatments. PMID- 18404564 TI - Isolation and structure determination of the cembranoid eunicin from a new genus of octocoral, Pseudoplexaura. AB - The known cembranoid diterpene eunicin (1) was isolated from a new genus of octocoral, Pseudoplexaura, collected in the Caribbean. The complete assignment of the 13C and 1H NMR spectra, including differentiation of pro-R and pro-S hydrogens and elucidation of all J values, was accomplished with the aid of TOCSY, HSQC, and HMBC data, and a consideration of three low-energy conformations using SYBYL. PMID- 18404565 TI - A new 3-arylcoumarin from the roots of an Egyptian collection of Lotus polyphyllos. AB - Further investigation of different fractions from Lotus polyphyllos Clarke roots resulted in the isolation of the new 3-arylcoumarin derivative; 4',6'-dihydroxy 7, 2'-dimethoxy-3-arylcoumarin (5). In addition, the known compounds beta sitosterol, 1-hexacosanol (1), n-tetracosyl p-coumarate (2), 4'-O-methylderrone (4), and quercetin (6) were identified. The structures were determined from spectroscopic data including 1D- and 2D-NMR experiments. PMID- 18404567 TI - Role of hypaphorine in the toxicity of Astragalus lusitanicus. AB - Hypaphorine, an alpha-N,N,N-trimethyltryptophan betaine, was isolated, for the first time, from Astragalus lusitanicus Lam. (Fabaceae), a plant highly toxic for lambs and goats. This alkaloid was characterized by NMR and MS analysis. Hypaphorine was previously reported to be a convulsive poison. To confirm the toxicity, it was synthesized and tested in goats. Hypaphorine was shown to be non toxic for goats even at a high dose of 2 g kg(-1) by oral administration. PMID- 18404568 TI - Acyclic phosphonate nucleotides and human adenylate kinases: impact of a borano group on alpha-P position. AB - Adenylate kinases are involved in the activation of antiviral drugs such as the acyclic phosphonates analogs PMEA and (R)PMPA. We examine the in vitro phosphorylation of PMEA and PMPA bearing a borano- or a H- group on the phosphorus atom. The alpha-borano or alpha-H on PMEA and PMPA were detrimental to the activity of recombinant human AMP kinases 1 and 2. Docking PMEA to the active site of AMP kinase 1 indicated that the borano group may prevent two conserved critical Arg interactions with the alpha-phosphate, resulting in substrate bad positioning. PMID- 18404569 TI - Diastereomer separation of azobenzene-tethered oligodeoxyribonucleotides and determination of their absolute configurations by enzymatic digestion. AB - Two diastereomers were produced by the introduction of azobenzene-tethering prochiral linker (2,2-bis(hydroxymethyl)propionic acid) in the modified ODN, which had been used for the photoregulation of DNA functions. We found that this modified ODN with sequence 5'-...pNpXpN...-3' (p = phosphate; N = nucleoside; X = azobenzene residue) could be digested to pX (the phosphate at the 5' side of X was left) by an over excess of Phosphodiesterase I. By comparing the retention time of pX from the separated diastereomer with that of authentic R- or S-pX on chiral HPLC, absolute configuration could be easily determined. PMID- 18404571 TI - A highly efficient and selective synthesis of 1,2,3-triazole linked saccharide nucleosides via "click chemistry". AB - A series of 1,2,3-triazole linked saccharide nucleosides were synthesized in high yield and selectivity via "click chemistry" of the 3'-azido-2'-deoxythymidine and the propargyl carbohydrates. [image omitted]. PMID- 18404570 TI - Novel N-3 substituted TSAO-T derivatives: synthesis and anti-HIV-evaluation. AB - Novel derivatives of the anti-HIV-1 agent, TSAO-T, bearing at the N-3 position alkylating groups or photoaffinity labels were prepared and evaluated for their anti-HIV activity. All of these compounds demonstrated pronounced anti-HIV-1 activity and inhibited HIV-1 RT; however, we were unable to detect stable covalent linkages between inhibitor and enzyme. In addition, compounds with an alcohol functional group connected to the N-3 position through a cis or trans double bond have been prepared. These compounds have been useful to study how the conformational restriction of the linker affects in the interaction between the N 3 substituent and the HIV-1 RT enzyme. PMID- 18404572 TI - ChipCheckII - predicting binding curves for multiple analyte strands on small DNA microarrays. AB - Incomplete binding, saturation, and cross-hybridization between partially complementary strands complicate the parallel detection of nucleic acids via DNA microarrays. Treating the competing equilibria governing binding to microarrays requires computational tools. We have developed the web-based program ChipCheckII that calculates total hybridization matrices for target strands interacting with probes on small DNA microarrays. The program can be used to compute the extent of cross-hybridization and other phenomena affecting fidelity of detection based on sequences, quantities of strands, and hybridization conditions as inputs. Enthalpy and entropy of duplex formation are generated locally with UNAfold, including those for complexes that are partially matched. Simulated binding versus temperature curves for portions of a commercial genome chip demonstrate the extent to which cross-hybridization can complicate DNA detection. ChipCheckII is expected to aid nucleic acid chemists in developing high fidelity DNA microarrays. PMID- 18404573 TI - De novo synthetic route to a combinatorial library of peptidyl nucleosides. AB - A stereoselective synthetic route has been developed for the combinatorial synthesis of a structurally unique class of C-4' side chain modified peptide linked nucleosides. The synthetic strategy and approach involves initial synthesis of a strategically functionalized amino butenolide template, utilizing L-serine as a chiral starting material. Subsequent transformation of the above lactone to C4' aminoalkyl substituted nucleosides, followed by the peptidic coupling of the C4' side chain amine with various amino acids completed the syntheses of the target peptidyl nucleosides. Employing the above route, and utilizing a combination of easily available nucleobases (4) and amino acids (6) as the two diversity elements, synthesis of a 24-member combinatorial library of the title peptide-linked nucleosides has been accomplished. PMID- 18404574 TI - Synthesis of 3'-acetamidoadenosine derivatives as potential A3 adenosine receptor agonists. AB - On the basis of high binding affinity of 3'-aminoadenosine derivatives 2b at the human A3 adenosine receptor (AR), 3'-acetamidoadenosine derivatives 3a-e were synthesized from 1,2:5,6-di-O-isopropylidene-D-glucose via stereoselective hydroboration as a key step. Although all synthesized compounds were totally devoid of binding affinity at the human A3AR, our results revealed that 3' position of adenosine can only be tolerated with small size of a hydrogen bonding donor like hydroxyl or amino group in the binding site of human A3AR. PMID- 18404575 TI - Regio- and diastereo-selectivity of montmorillonite-catalyzed oligomerization of racemic adenosine 5'-phosphorimidazolide. AB - Clay is a possible candidate for an effective catalyst in prebiotic chemical evolution of biomolecules. Montmorillonite was reported to effectively catalyze oligomerization of racemic adenosine 5'-phosphorimidazolide (DL-ImpA). In the oligomerization reaction, considerable amounts of cyclic dimers as well as linear dimers were produced in the oligomerization reactions. To assess the regio- and diastereo-selectivities of the oligomerization reaction, the dimer products including cyclic dimers were completely identified by means of enzymatic degradation reactions of the products. PMID- 18404577 TI - Mosaicism, modules, and the evolution of birds: results from a Bayesian approach to the study of morphological evolution using discrete character data. AB - The study of morphological evolution after the inferred origin of active flight homologous with that in Aves has historically been characterized by an emphasis on anatomically disjunct, mosaic patterns of change. Relatively few prior studies have used discrete morphological character data in a phylogenetic context to quantitatively investigate morphological evolution or mosaic evolution in particular. One such previously employed method, which used summed unambiguously optimized synapomorphies, has been the basis for proposing disassociated and sequential "modernizing" or "fine-tuning" of pectoral and then pelvic locomotor systems after the origin of flight ("pectoral early-pelvic late" hypothesis). We use one of the most inclusive phylogenetic data sets of basal birds to investigate properties of this method and to consider the application of a Bayesian phylogenetic approach. Bayes factor and statistical comparisons of branch length estimates were used to evaluate support for a mosaic pattern of character change and the specific pectoral early-pelvic late hypothesis. Partitions were defined a priori based on anatomical subregion (e.g., pelvic, pectoral) and were based on those hypothesized using the summed synapomorphy approach. We compare 80 models all implementing the M(k) model for morphological data but varying in the number of anatomical subregion partitions, the models for among-partition rate variation and among-character rate variation, as well as the branch length prior. Statistical analysis reveals that partitioning data by anatomical subregion, independently estimating branch lengths for partitioned data, and use of shared or per partition gamma-shaped among-character rate distribution significantly increases estimated model likelihoods. Simulation studies reveal that partitioned models where characters are randomly assigned perform significantly worse than both the observed model and the single-partition equal-rate model, suggesting that only partitioning by anatomical subregion increases model performance. The preference for models with partitions defined a priori by anatomical subregion is consistent with a disjunctive pattern of character change for the data set investigated and may have implications for parameterization of Bayesian analyses of morphological data more generally. Statistical tests of differences in estimated branch lengths from the pectoral and pelvic partitions do not support the specific pectoral early-pelvic late hypothesis proposed from the summed synapomorphy approach; however, results suggest limited support for some pectoral branch lengths being significantly longer only early at/after the origin of flight. PMID- 18404578 TI - [Iatrogenic injuries - avoiding errors in orthopaedics and traumatology]. PMID- 18404579 TI - [Risk management in hospitals: standard operating procedures in aviation as a model for structuring medical communication]. AB - AIM: The working conditions for medical staff in German hospitals with emergency units are characterised by stress peaks. In such moments mistakes are more likely to be made. Communication problems and misunderstandings caused by these settings are the most frequent causes for blunders, errors and mistakes. The aim is to show an easy way to reduce the incidence of these problems. METHOD: Within this prospectively randomised trial we compare the ability of 50 physicians to remember standardised information on patient's data passed on to them in a briefing. Half of the subjects used the so called 5-finger method as an easy means of memorising standardised information. The other 25 physicians memorised the patient's data in the usual way. In order to assess the information recalled by the subjects, all of them were asked to answer 15 questions on the data they had been given an hour earlier. RESULTS: The subjects who used the 5-finger method were able to give on average 9.88 (of 15) correct answers and thus remembered significantly (p = 0.031) more information than the members of the control group. CONCLUSION: Communication errors caused by homophony, ambiguity, anticipation and mental models are can be avoided in well-structured communication situations. The rules for such a type of communication can easily be learned and applied. Standardised communication has already proved to reduce the error rates in aviation. We conclude that it can be helpful to introduce standardised communication rules such as the 5-finger method in medical communication. PMID- 18404580 TI - [Complications after hip resurfacing using the ASR prosthesis in patients with osteoarthritis]. AB - PURPOSE: The purpose of this study was to evaluate peri- and postoperative complications and the clinical/radiological results after hip resurfacing with the ASR prosthesis in patients with osteoarthritis. METHOD: A prospective cohort study was performed including all patients who received a hip resurfacing (ASR prosthesis, DePuy, Motech, Warsaw, IN, USA) because of osteoarthritis between 1/2005 and 7/2006. 115 prostheses were performed in 110 patients (58 female, 52 male). In all patients a dorsal approach was used. The mean age was 57.82 years (range: 34 to 72 years) and the mean follow-up was after 12.5 months (range: 6 to 23 months). At the time of follow-up an X-ray (a. p. and axial) of the operated hip was performed in addition to the to standard clinical examination. To measure the functional outcome the Harris hip score was used. RESULTS: We observed postoperative complications in 18 patients. Three non-trauma-related femoral neck fractures were seen after a mean of 58 days (range: 41 to 70 days). One trauma related femoral neck fracture was found after 7 months. Neural complications were found in two patients. In 8 patients a revision of the acetabular component was necessary. 3 dislocations could be managed without further revision. One revision was necessary because of a persisting seroma without infection. No infections or thromboembolic complications were seen. The mean Harris hip score developed from preoperative 59 points (range: 50 to 65 points) to postoperative 96 points (range: 85 to 100 points). CONCLUSIONS: Excellent early postoperative results can be reached by hip resurfacing. Special attention is necessary performing the correct positioning of the femoral and acetabular components. If notching is seen intraoperatively the operation should be switched and a large head prothesis should be implanted. Long-term results are still lacking. PMID- 18404581 TI - [Screw-in performance of threaded cups]. AB - INTRODUCTION: For threaded cups, cup diameter, the shape of the threads and the shape of the cup have a major influence on the screw-in performance. The designs of standard implants vary greatly, making it difficult to carry out comparative studies to provide clear-cut results on the effect of individual parameters on the insertion process. MATERIAL AND METHODS: 15 different prototype cups were manufactured for this study. Three sizes, three shapes and five thread designs were tested using artificial bone. Screw-in performance was measured with the slope of insertion torque before and after reaching the seating point, the torque at the seating point, the insertion angle and the change of slopes at the seating point. RESULTS: Modified trapezoid threads have the lowest insertion resistance and the clearest seating point as well as the most distinct change of slope at the seating point. Flat threads show lower insertion resistance and more increased slopes at the seating point than sharp threads. The slope of insertion torque before and at the seating point is higher for sharp threads in comparison to the other threads, the change of slop is however low. Larger cups show a higher insertion resistance. CONCLUSION: Screw-in performance is decisively influenced by the design of the threads. Modified trapezoid threads are the easiest to insert with excellent manual control, thus reducing the risk of intra operative overturning. Flat threads also have a low screw-in resistance. Sharp threads have an unsatisfactory screw-in performance because the seating point cannot be "felt". PMID- 18404582 TI - [Radiological mid-term results of total knee arthroplasty with femoral components of different materials]. AB - AIM: The present work aimed at evaluating the radiological mid-term results of femoral components of different materials in an identically designed total knee system. METHOD: 31 knees with zirconium femoral components and 32 standard femoral components were evaluated about 4 years following surgery using the Roentgenographic Evaluation and Scoring System of the Knee Society. RESULTS: In both groups, the implant position was correct. There was a statistically significant higher rate of radiolucent lines in zones 1 and 4 at the tibia site in the zirconium group. CONCLUSION: Our clinical results disagree with the results of biomechanical studies of the same type of prosthesis. However, there are some findings from animal studies showing that zirconium ions may inhibit the mineralisation of osteoid, which could be an explanation for the higher rate of radiolucent lines seen in the present study. PMID- 18404583 TI - [Influence of the surgical approach on postoperative rehabilitation after TKA]. AB - BACKGROUND: Minimising the soft tissue trauma and early rehabilitation are among the major aims using the MIS technique in joint replacement. AIM: The aim of this prospective randomised study is to compare the results after TKA using an MIS approach versus a standard approach. METHOD: We compared 30 TKA using a mini midvastus approach (MIS group) with 30 conventionelly performed TKA using a midvastus approach (control group). In all cases the same implants (NexGen LPS) were used. The Knee Society score (KSS), an activity score, the visual analogue scale, myoglobin and creatinine kinase as well as the blood loss were measured up to 12 weeks postoperatively. Implant positioning was evaluated using the Knee Society Roentgenographic evaluation and scoring system. RESULTS: Advantages of the MIS group were measured in KSS and activity score up to 12 weeks postoperatively. The increase of myoglobin and creatinine kinase was lower in the MIS group up to 24 hours or 72 hours, respectively. No differences were found comparing the implant positioning and blood loss. CONCLUSION: The MIS technique in TKA with minimisation of the soft tissue trauma leads to better clinical and functional results in the early postoperative time and does not modify the implant positioning. PMID- 18404584 TI - [Shoulder resurfacing in patients with rotator cuff arthropathy and remaining subscapularis function]. AB - INTRODUCTION: Glenohumeral arthritis with a massive rotator cuff tear is a devastating condition that seriously compromises the comfort and function of the shoulder. Cuff tear arthropathy presents a unique surgical challenge and many arthroplasty options were used for its treatment. The purpose of this study was to evaluate the clinical and radiological results of Copeland cementless surface replacement arthroplasty (CSRA) applied in patients with cuff tear arthropathy and intact subscapularis function. PATIENTS AND METHODS: The study was conducted on twenty-five shoulders in twenty-five patients with cuff tear arthropathy with the subscapularis tendon still intact. The patients were prospectively followed up clinically and radiologically for a mean of 26 months (range: 15 - 38 months). There were 16 female and 9 male patients. The mean age was 69.04 years (range: 53 - 83 years). The mean operative time was 38 minutes (range: 28 - 56 minutes). The clinical assessment was performed with the Constant score. RESULTS: The Constant score significantly improved from a mean of 14.04 points preoperatively to 53.17 points postoperatively. Of the patients, 88 % considered the shoulder to be much better or better as a result of the operation. Radiologically, the humeral offset, the lateral glenohumeral offset (coracoid base to the greater tuberosity), height of centre of instant rotation and the acromiohumeral distance were significantly increased. No intra- or postoperative complications were encountered. CONCLUSION: Our early results with the use of Copeland surface replacement in selected cases with cuff tear arthropathy are encouraging. The patients showed significant clinical (pain and range of motion) and radiological improvements. Moreover, if the surface replacement were to fail for any reason, it can be revised to a reverse prosthesis type as there is no lack of bone stock. PMID- 18404585 TI - [Reconstruction of the rotational centre of the humeral head depending on the prosthetic design]. AB - AIM: The displacement of the rotation centre of the humeral head in relation to the axis of the shaft varies strongly between individuals. The Affinis shoulder endoprosthesis has a double excentric adjustment possibility that permits us to adapt the head to the medial and dorsal offset. So far, such examinations have taken place exclusively on anatomic preparations. This raises the question of whether the need for such a prosthetic system can be derived from the anatomic variation of the pivot points. METHOD: In 49 patients with an implanted Affinis shoulder prostheses, we calculated the individual rotation centres of the head from the position of the relocatable prosthetic cone and the excentric position of the head. In addition, we used the Constant score to record the clinical function. RESULTS: In the examined case material, we needed the entire setting range of 12 mm mediolateral and 6 mm dorsoventral of the prosthesis, as it presented a great variation of the rotational centres of the head. The majority of the head centres were displaced in a posterior-lateral direction. The examination showed that none of the found anatomic head centres could have been reconstructed exactly with a conventional prosthesis. Prostheses with a single excentricity would allow correct adjustment in only 9 cases, as the adjustable rotational centres of the head are situated in an orbit, which limits the setting possibilities. In 20 of the 49 patients humeral head prosthesis were implanted in cases of primary osteoarthritis. The Constant score of this group improved from 18 points preoperatively to 76 points after 24.1 months in the mean. CONCLUSION: The high variation of the head centres indicates the necessity of a freely adjustable system, such as in a double excentric bearing. This is a way that permits an optimal reconstruction of the anatomic conditions. Therefore all technical chances for bony balancing should be used to adapt the prosthesis to the bone and not the bone to the prosthesis. PMID- 18404586 TI - [Clinical results after replacement of comminuted radial head fractures (Mason III and IV) by the bipolar radial head prosthesis of Judet]. AB - AIM: Long-term results after resection of comminuted fractures of the radial head (Mason III and IV) may lead to valgus elbow instability, radius proximalisation and rotatory instability. Radial head replacement has been used to prevent and treat these complications. The aim of this study was to define the value of the bipolar radial head prosthesis of Judet for treating comminuted fractures of the radial head. METHOD: Over a 5-year period, 14 radial head prosthesis were implanted in 12 patients. Retrospectively, we studied the clinical and radiological results. RESULTS: Eleven patients with thirteen implants were analysed at a mean follow-up of 33.4 +/- 20.4 months. In all patients, the elbow was stable. Subjectively, we found good and excellent results with one exception. Compared to the pre-trauma status, the subjective rate was 78 +/- 12 %. According to the score of Radin and Riseborough, five of the results were found to be good and eight to be fair. According to the Broberg and Morrey score, one result was found to be very good, seven to be good and five to be fair. The mean DASH score was 10.2 +/- 10.1 points. Two temporary nerve lesions were observed. CONCLUSION: In comminuted fractures of the radial head (Mason III and IV), bipolar radial head replacement with the Judet prosthesis leads subjectively to very good and good and functionally to good and fair results on the medium-term view. Joint stability is achieved and secondary complications like valgus elbow deformity and proximal radial migration are prevented. Patients must be informed about the possibility of temporary nerve lesions, heterotopic ossification causing limitation of motion and the lack of long-term results. PMID- 18404588 TI - [2D-fluoroscopic navigated screw osteosynthesis of acetabular fractures: A preliminary report]. AB - INTRODUCTION: The current gold standard for operatively treated acetabular fractures is open reduction and internal fixation. In this study we report the early results of percutaneous screw osteosynthesis of acetabular fractures in 14 cases, using a computer navigation system to reduce screw misplacement, approach associated risks and radiation exposure. RESULTS: All 36 acetabular screws were placed correctly without any perioperative complication. The mean operation time per screw was 54 min, the mean fluoroscopic time per screw was 87 s. In the follow-up examinations (current rate: 71 %, mean follow-up: 13.1 +/- 1.8 months) no fixation failure or development of post-traumatic osteoarthritis was seen. One wound infection occured. CONCLUSION: Our first experience with this novel approach is promising and serious advances of fluoroscopic navigated percutaneous screw osteosynthesis of well selected acetabular fractures can be expected. PMID- 18404587 TI - [Parkinson's disease influences the perioperative risk profile in trauma patients]. AB - AIM: Parkinson's disease (PD) is one of the most common degenerative diseases of the central nervous system affecting elderly patients with increasing demographic prevalence. The aim of this study was to define the perioperative risk profile in trauma patients suffering from Parkinson's disease in order to improve treatment options in these patients. METHOD: Over a period of 13 years, 16 patients suffering from Parkinson's disease treated in the department of trauma surgery were retrospectively compared using matched-pair analysis with 16 controls not affected by PD. Both groups of patients were assessed regarding morbidity, length of treatment and rehabilitation. RESULTS: Trauma patients suffering from Parkinson's disease showed an increase in morbidity risk. Postoperative falls occurred significantly, infections of the urinary tract and pneumonia tendentiously more often in PD patients. Postoperative stay and length of treatment were significantly longer in patients with PD. These patients also tended to stay longer preoperatively and remained longer in intensive care. PD patients required on-ward rehabilitation significantly more often. CONCLUSION: Concomitant Parkinson's disease is a significant factor of perioperative morbidity in trauma patients. Perioperative morbidity in PD patients may be influenced by early diagnostic and therapeutic measures. PMID- 18404589 TI - [The small-fragment double plate osteosynthesis in C1 to C3 fractures of the tibial pilon]. AB - Intra-articular fractures of the tibial pilon always have been a challenge for the surgeon and his operative technique. With conservative or operative management, we have to take care of destroyed articular surfaces and traumatised soft tissues. Therefore, tibial pilon fractures are known in the literature as complicated lesions with a high rate of complications. Between 1993 and 2001 twenty-one patients who were between 18 and 65 years old and suffered a monotrauma of the tibial pilon of the type C1 to C3 of the AO classification, were treated with a staged reconstruction. Stabilisation was done by small fragment double plate osteosynthesis. In this retrospective study, we could review 17 patients clinically and radiologically and assess their outcome with the ankle hindfoot and the Bargon scores. In accordance with the ankle hindfoot score, there were 11 excellent and good results, 5 moderate and one poor. In accordance with the Bargon score, 41 % of the patients had a severe arthrosis, 24 % a moderate and 35 % a light to no arthrosis. Our results show that the small fragment double plate osteosynthesis offers a high stability, allows careful soft tissue handling and can be done with a low ratio of complications. Long-term results are gratifying, although the radiological examinations often show a certain degree of arthrosis of the ankle joint. PMID- 18404590 TI - [Reliability of digital measurement of acetabular index in hip dysplasia to the time children start walking]. AB - AIM: An assessment of the reproducibility of the acetabular index on the basis of anteroposterior radiographs of the pelvis was performed in order to rule out residual dysplasia of the hip in the second year of life. METHOD: 51 consecutive patients (41 girls, 10 boys) with congenital dysplasia of the hip (35 bilateral, 16 unilateral) were included. Approximately at the time children started walking (mean age: 13.9 months; range: 10 - 22 months), digitised anteroposterior radiographs of the pelvis were recorded. Acetabular indices were measured bilaterally and blinded by two observers (O1 and O2), then classified according to the normal values of the hip joint described by Tonnis et al. (normal = lower single standard deviation [s], light dysplasia = above single standard deviation [s - 2 s] and severe dysplasie = above double standard deviation [2 s]). O2 repeated a second set of observations 6 weeks after the first. 306 values of the acetabular index (O1: 102 values; O2: 204 values) were statistically evaluated. Reliability was expressed by Pearson's correlation coefficient (PCC) and 95 % confidence interval. The rater agreement of the classification into normal values of the hip joint was calculated with the kappa coefficient. RESULTS: The PCC for intraobserver/interobserver reliability was 0.928/0.875. The 95 % confidence interval for intraobserver reliability was +/- 3.1 degrees and for interobserver reliability - 3.1 degrees /+ 5.9 degrees . A comparison of the first series of measurements by O1 with the series of measurements by O2 presented congruence in 66 of 102 hip joints classified according to the normal values of the hip joint as described by Tonnis. In 36 hip joints the classification was deviant, in 15 the classification differed between normal and s, in 21 between s and 2 s. The kappa coefficient of 0.51 showed moderate interobserver agreement. CONCLUSION: The reproducibility of radiological surveillance in congenital hip dysplasia, approximately to the time children start walking, can be considered as adequate. Correct positioning of the patient and exact definition of radiological landmarks are essential. In spite of good statistical values for reliability, decreasing accuracy in classifying into Tonnis' normal values of the hip joint is seen with increasing degree of dysplasia. In marginal cases, before coming to the decision for operative treatment, a second reading, ideally by the same observer and with a chronological shift, is advisable. PMID- 18404591 TI - [Comparison of the treatment of slipped capital femoral epiphysis with K-wires and cannulated titanium screws]. AB - AIM: The treatment of slipped capital femoral epiphysis (SCFE) is usually treated operatively, but there is still no consensus about the method to be used. Up to a 30 degrees degree of slipping, the epiphysis is normally fixed in situ. The aim of our study was to compare the intermediate results after fixation in situ by K wires versus cannulated titanium screws (Konigsee-Implantate, Konigsee-Aschau, Germany). METHODS: In this study 46 patients with SCFE grade I and II and mostly chronic slipping of the epiphysis were included. After fixation in situ and, if necessary, careful, closed reposition, the patients were clinically and radiologically followed-up for one year. The clinical results were documented by the score adapted from Heyman and Herndon. Furthermore, MRI scans were done to evaluate the vitality of the epiphysis pre- and postoperatively, when titanium screws were used. RESULTS: Clinical follow-up showed comparable results in the clinical scores after fixation by K-wires or cannulated titanium screws (3.13 +/- 1.02 vs. 3.10 +/- 1.01). After the treatment with titanium screws we saw a higher rate of abnormal gait (33.3 % vs. 19 %), a decreased rate of the positive Drehmann sign (10 % vs. 38 %) and a lower rate of revisions (16 % vs. 50 %) in comparison to K-wire fixation. After displacement of the K-wires we saw chondrolysis and prearthrosis in one case. Removal of the K-wires was done without any complications, while the removal of the cannulated titanium screws failed in 4 of 10 cases. CONCLUSION: The treatment of SCFE with K-wires and cannulated titanium screws showed comparable results in the clinical follow-up. The treatment with cannulated titanium screws reduces the number of necessary revisions, but the removal of the material is hindered. Because of the lower rate of complications we prefer in the meantime the use of cannulated steel screws. PMID- 18404592 TI - [Embolisation of aneurysmal bone cysts as a minimally invasive treatment option - presentation of two cases]. AB - AIM: The aim of this study was to evaluate embolisation as a therapy option for aneurysmal bone cysts of the trunk. METHOD: Case reports about two males with intermittent pseudo-radicular lumboischialgia and coxalgia are discussed. RESULTS: The diagnostic work-up and biopsies verified an aneurysmal bone cyst in both males. In one patient the tumour-like lesion was localised in the fifth lumbar vertebral body, in the other in the left ischium and pubis. Arterial embolisation was performed in both cases. Follow-up at 6 and 24 months after embolisation showed a significant increase of sclerosis and a reduced volume of the cysts. CONCLUSION: In accord with literature data, arterial embolisation seems to be a sufficient and minimally invasive therapy option in aneurysmal bone cysts of the spine and the pubis. PMID- 18404593 TI - [The "well-leg" syndrome: calf compartment after lithotomy position]. AB - AIM: The lower leg compartment syndrome is in most cases a post-traumatic soft tissue complication. Nevertheless, it can also occur in a primarily not injured calf, due to external muscle compression during lithotomy positioning. This complication is seen rarely after gynaecological, urological or abdominal operations performed in the Lloyd-Davis position. METHOD: We present a case of calf compartment after prolonged lithotomy position of the uninjured leg during the nailing of a femur shaft fracture. RESULTS: Postoperatively a diffuse turgor of the healthy calf was seen. The compartment pressures were elevated and the enzyme creatine kinase was elevated up to 3000 U/l. After dermatofasciotomy the enzyme level normalised, but in the further course we had to resect the necrotic peroneal muscle compartment. CONCLUSION: During operative procedures at the coxal femur, it is necessary to deposit the other leg in the lithotomy postion. By this means, the compartment pressures can increase in the calf muscles to critical values. Together with operation times longer than 4 hours and a high ponderosity a bedding compartment can develop. As it is in the assumed "healthy" leg, this syndrome can be overseen, especially in sedated patients. It has to be considered with high creatine kinase levels and the typical risk factors. If a compartment syndrome is suspected, we recommend early dermatofasciotomy, as this is a low risk and low-complication operation, in comparison to a later performed compartment decompression. By extending both legs, especially in risky patients, the surgeon can avoid this dangerous complication. PMID- 18404595 TI - A time course analysis of systemic administration of aqueous licorice extract on spatial memory retention in rats. AB - In the present study, the time course of the effects of Glycyrrhiza glabra L. (Leguminosae) aqueous extract (GE), administered systemically to rats, on the spatial memory retention in the Morris water maze was investigated. The dose of glycyrrhizin (GL), i. e., 0.5, 2.5 and 5 mg/mL in daily water intake of GE was administered to three groups of rats. The first, second and third groups received GE for 1, 2 and 4 weeks, respectively (each group included 3 subgroups). Three additional control groups of animals received only tap water during the same periods of time. After terminating the treatments, all animals were trained for four days; each day included one block and each block contained four trials. Test trials were conducted 48 h after the completion of the training period. Nicotine (1 microg/side) was infused into the CA1 region of the hippocampus as a positive drug control. GE treatment decreased both escape latency and traveled distance, but not swimming speed, compared with control, suggesting significant spatial memory retention enhancement by GE. Statistical analysis did not show any significant difference between GE-treated animals and the nicotine group in escape latency and traveled distance. At the end of the testing trials plasma samples were collected and the concentrations of glycyrrhetinic acid (GA) as a major metabolite of GL were measured in the different groups of treated rats. The maximum concentration was observed after four weeks of GE administration at 5 mg/mL of GL. These results showed that the enhancement effect of GE on spatial memory retention does not correlate with GA blood levels. PMID- 18404596 TI - Protective effects of amburoside A, a phenol glucoside from Amburana cearensis, against CCl4-induced hepatotoxicity in rats. AB - The aim of this study was to investigate the possible beneficial effects of amburoside A, AMB [4-(O-beta- D-glycopyranosyl)benzyl protocatechoate], against carbon tetrachloride (CCl (4)) toxicity in rats. AMB is a phenol glucoside from the Brazilian medicinal plant Amburana cearensis, popularly used for the treatment of respiratory tract affections. Acute AMB (25 and 50 mg/kg, I. P. or P. O.) treatments of CCl (4)-intoxicated rats significantly inhibited the increase in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, as compared to the group treated with CCl (4) only. Histological studies showed less centrolobular necrosis and inflammatory cell infiltrates in the liver of animals treated with AMB plus CCl (4), when compared to the group treated with CCl (4) alone. In hepatic tissues, AMB at both doses inhibited CCl (4)-induced thiobarbituric acid-reactive substances (TBARS) formation, indicating a blockade of CCl (4)-induced lipid peroxidation. AMB also reversed the decrement in glutathione contents of hepatic tissues in CCl (4)-intoxicated rats. Furthermore, it restored catalase activity to normal values, which was significantly increased after CCl (4) treatment. Our results indicate that CCl (4)-induced oxidative damage in hepatic tissues is reversed by AMB treatment. The protective effect of AMB is probably due to the phenolic nature of this glucoside. PMID- 18404597 TI - Plant polyphenols against UV-C-induced cellular death. AB - The glycosylated phenylpropanoid verbascoside isolated from cultured cells of the medicinal plant Syringa vulgaris (Oleaceae) has previously been characterized as an effective scavenger of biologically active free radicals such as hydroxyl, superoxide, and nitric oxide, as a chelator of redox active transition metal ions (Fe (2+), Fe (3+), Cu (2+), and Ni (2+)), and an inhibitor of lipid peroxidation. In the present work, we have compared the cytoprotective effects of the biotechnologically produced verbascoside with two commercially available polyphenols (the glycosylated flavonoid rutin and its aglycone quercetin) against free radical-mediated UVC-induced cellular death in cultures of human keratinocytes (HaCaT) and breast cancer cells (MCF 7). We have shown that all the polyphenols studied afforded effective protection against UVC-induced necrosis and did not prevent UVC-induced apoptosis in both normal and tumor cell lines. The cytoprotection did not correlate either with UVC absorbance by polyphenols or with their superoxide radical scavenging properties. However, UVC protection strongly depended on the lipid peroxidation inhibiting and Fe (2+) chelating properties of polyphenols. We suggest that these plant polyphenols could be feasible for a photoprotection of human skin. PMID- 18404598 TI - Ghrelin as an acute-phase reactant during postoperative stress response. AB - Ghrelin is a growth hormone-releasing peptide, discovered in 1999 by Kojima et al. Its potential role in inflammation and stress response is not yet clear. The purpose of this study was to characterize perioperative levels of circulating ghrelin in relation to different surgical procedures. The authors compared plasma ghrelin changes with cortisol, cytokines, and acute-phase proteins. The prospective study was performed on 22 patients with resection for colon cancer (group 1). Group 2, functioning as a comparative group, consisted of 22 patients with elective laparotomic cholecystectomy. Plasma concentrations of ghrelin, cortisol, tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta, IL-6, IL-8, soluble IL-2 receptor, C reactive protein, and alpha1-antitrypsin were estimated repeatedly during a 72-hour postoperative period. Data revealed significant elevation of plasma ghrelin 24 hours after resection of coli (median 508.0 ng/l, interquartile range 398.2-633.7 ng/l) in relation to both preoperative levels (317.6 ng/l, 253.4-355.1 ng/l, p<0.01) and group 2 maximal postoperative levels (386.2 ng/l, 324-432 ng/l, p<0.05). Ghrelin levels returned to initial status 36 48 hours after surgery with subsequent decline to subnormal levels. The regression coefficient was the highest for ghrelin and TNF-alpha 24 hours after laparotomy (r=0.64, p<0.05) and for ghrelin and IL-6 24 hours after surgery (r=0.56, p<0.05). Maximal postoperative levels of all tested parameters except for cortisol and IL-1beta differed significantly between both patient groups at p<0.05. After large abdominal surgery, ghrelin shows itself as an acute-phase reactant. The significant correlation between ghrelin and inflammatory cytokines supposes their regulatory role in this period. Our comparison of more- and less invasive surgical procedures with similar nutritional restrictions argues for a dominant role of inflammatory factors in postoperative ghrelin elevation. PMID- 18404599 TI - Antiatherogenic effects of cilostazol and probucol alone, and in combination in low density lipoprotein receptor-deficient mice fed with a high fat diet. AB - Cilostazol, an antiplatelet drug, and probucol, a cholesterol-lowering drug, are reported to ameliorate atherosclerosis in animal models. However, their combined effect on atherosclerosis is unclear. We therefore evaluated their combined effect on atherosclerotic lesions in LDL receptor-deficient mice. Male LDL receptor-deficient mice were fed a high fat diet with or without cilostazol alone, probucol alone, or with cilostazol and probucol in combination, for 8 weeks. Body weight and plasma lipid levels were measured before and during treatment. At the end of treatment, the size distribution of plasma lipoproteins was analyzed by HPLC and then plasma HDL cholesterol levels and en face aortic atherosclerotic lesion areas were measured. Probucol alone significantly decreased both total cholesterol and HDL cholesterol, while cilostazol alone did not decrease total cholesterol, but significantly increased HDL cholesterol. Both cilostazol alone and probucol alone significantly decreased atherosclerotic lesion areas, and their combined administration showed more significant decreases than when each drug was administered singly. The combination of cilostazol and probucol was more effective in preventing atherosclerotic lesion formation than the administration of each drug alone; this may provide us with a new strategy for treating atherosclerosis. PMID- 18404600 TI - Transluminal closure for NOTES: an ex vivo study comparing leak pressures of various gastrotomy and colotomy closure modalities. AB - BACKGROUND AND STUDY AIMS: Transluminal closure is fundamental to the safe introduction of natural orifice transluminal endoscopic surgery (NOTES) into humans. Suture, staples, and clips have been used. We aimed to evaluate the acute strength of various gastrotomy and colotomy closure techniques in an ex vivo porcine model by assessing air leak pressures. PATIENTS AND METHODS: The following closure modalities were assessed with at least five samples per arm: conventional open/laparoscopic suturing techniques including full-thickness interrupted sutures, double-layer sutures, and purse-string sutures, as well as endoscopic clips and endoscopic staples. Historical values for transgastric closures with hand-sewn interrupted sutures, endoscopic clips, and a prototype endoscopic suture device were used from our laboratory's prior study. RESULTS: Using Kruskal-Wallis analysis, the overall comparisons were significant ( P = 0.0038 for gastrotomy closure; P = 0.0018 for colotomy closure). Post hoc paired comparisons revealed that the difference between all closure arms versus negative control were significant. Significance could not be established among the various closure arms. However, trends suggested hand-sewn double-layer sutures, endoscopic staples, and both hand-sewn and endoscopically-placed purse-string sutures produced the strongest closures. Furthermore, endoscopic clips appeared sufficient for colotomy closure when ideally placed. CONCLUSIONS: Suture (both hand-sewn and endoscopically deployed) appears to produce the strongest closures in both stomach and colon, with the important caveats that (1) a continuous through-thickness suture track be avoided, such as in the full-thickness closure, or (2) suture holes be buried, such as in the purse-string configuration. When suture tracks are full-thickness, they can serve as leak sites. Staples and clips can produce comparable closures, but only under ideal conditions. PMID- 18404601 TI - Methylene blue-aided cholangioscopy in patients with biliary strictures: feasibility and outcome analysis. AB - BACKGROUND AND STUDY AIMS: Chromoendoscopy using methylene blue is employed in the gastrointestinal tract to delineate neoplastic lesions. We tested the value of chromoendoscopy during choledochoscopy for characterization of local inflammation, neoplasias, and other alterations in patients with biliary strictures. METHODS: Patients with suspected biliary lesions were scheduled for endoscopic retrograde cholangiography with subsequent cholangioscopy. After initial inspection of the bile duct, 15 ml methylene blue (0.1 %) was administered via the working channel of the cholangioscope. Newly appearing circumscribed or unstained lesions were judged according to their macroscopic type and staining features. Methylene-blue-aided diagnosis was compared with either clinical follow-up of the patients or, in some cases, with the results of targeted biopsies. RESULTS: A total of 55 patients [biliary stenosis/cholestasis of unknown origin (n = 24), stenosis after orthotopic liver transplantation (n = 11), primary sclerosing cholangitis (n = 20)] were included. Methylene blue unmasked subtle mucosal changes and permitted macroscopic characterization of circumscribed lesions. Characteristic surface staining patterns were seen in chronic inflammation, dysplasia, and ischemic-type biliary lesions. Nondysplastic mucosa appeared homogeneously stained, whereas scarred strictures showed a weak uptake of methylene blue. CONCLUSION: In this prospective feasibility study, methylene-blue-aided cholangioscopy was used for the first time to define different staining patterns of the bile duct. The differences in staining patterns identified normal, dysplastic, and inflamed mucosa of the bile duct, as was proved by follow-up or, in some cases, histology. Whereas homogeneous staining predicted the presence of normal mucosa, absence of staining of circumscribed lesions, or diffused staining of such lesions, represented neoplastic changes or inflammation. PMID- 18404603 TI - Changing dietary sodium alters the chronic cardiovascular effects of losartan in rats. AB - INTRODUCTION: We have previously demonstrated a profound hypotensive response to the angiotensin II type 1 (AT1)-receptor antagonist losartan in rats consuming a normal salt diet that is not seen in salt-loaded rats, presumably due to a suppression of the renin-angiotensin system (RAS) by high sodium levels. The purpose of the present study was to examine the cardiovascular effects of changing dietary sodium intake during chronic treatment with losartan. We hypothesised that during blockade of AT1-receptors by chronic losartan infusion, when renin levels would be elevated regardless of dietary sodium, changing diets from high to normal or normal to high salt would have no effect on mean arterial pressure (MAP). MATERIALS AND METHODS: To test this hypothesis, groups of rats instrumented with radiotelemetry transducers for MAP monitoring and venous catheters for infusion were initially placed on either a 0.4% salt content diet, referred to as Losartan Normal diet - High salt diet (LosN-HI, n = 7), or a 4.0% salt content diet, referred to as Losartan High salt diet - Normal diet (LosHI-N, n = 9). After a thee-day control period, infusion of losartan was begun in all rats (10 mg/kg/day in 7 ml/day isotonic saline i.v.). After 10 days, diets were switched between groups and data were collected for another 10 days, after which losartan infusion was terminated for a 10-day recovery period. RESULTS: At the start of losartan infusion MAP was observed to be similar between LosN-HI rats (101+/-2 mmHg) and LosHI-N rats (101+/-2 mmHg). By day seven of the first 10 day protocol, MAP in LosN-HI rats had fallen to 71+/-4 mmHg while decreasing to 90+/ 2 mmHg in LosHI-N rats. Five days after switching diets, MAP in LosN-HI rats had risen back to 85+/-3 mmHg, while MAP in LosHI-N rats had fallen to 75+/-2 mmHg. CONCLUSIONS: These results do not support our hypothesis, suggesting that changing dietary sodium can alter the chronic hypotensive response to losartan regardless of the initial state of the RAS. PMID- 18404602 TI - Concomitant calcium entry blockade and inhibition of the renin-angiotensin system: a rational and effective means for treating hypertension. AB - Pharmacological treatment of hypertension is effective in preventing cardiovascular and renal complications. Calcium antagonists (CAs) and blockers of the renin-angiotensin system [angiotensin-converting enzyme (ACE) inhibitors and angiotensin II antagonists (ARBs)] are widely used today to initiate antihypertensive treatment but, when given as monotherapy, do not suffice in most patients to normalise blood pressure (BP). Combining a CA and either an ACE inhibitor or an ARB considerably increases the antihypertensive efficacy, but not at the expense of a deterioration of tolerability. Several fixed-dose combinations are available (CA + ACE-inhibitors: amlodipine + benazepril, felodipine + ramipril, verapamil + trandolapril; CA + ARB: amlodipine + valsartan). They are expected not only to improve BP control, but also to facilitate long-term adherence with antihypertensive therapy, thereby providing maximal protection against the cardiovascular and renal damage caused by high BP. PMID- 18404604 TI - Angiotensin type 2 receptor is expressed in human atherosclerotic lesions. AB - OBJECTIVE: Expression of the angiotensin type 2 receptor (AT2-receptor) occurs in many animal models of atherosclerosis. However, its expression in human plaques and its functional role remains undetermined. This study examined AT2-receptor expression in human atherosclerotic plaque and also explored its potentially important functional role in atherosclerosis. MATERIAL AND METHODS: We analysed carotid atherosclerotic plaques obtained from 14 Caucasian patients who had previously undergone endarterectomy for symptomatic carotid artery stenosis. Half of all subjects received treatment with an angiotensin receptor blocker (ARB) (n=7); the remaining subjects received no intervention in the renin-angiotensin system (n=7). Immunohistochemistry measured tissue expression of smooth muscle cells (a-actin), macrophages (CD68 antibody), collagen (picro-sirius), and AT2 receptor (AT2-receptor antibody). RESULTS: AT2-receptor expression occurred consistently in all specimens. Although cellular localisation varied, AT2 receptor expression levels correlated with macrophage levels (p<0.01). Compared to conventional treatment, ongoing ARB treatment affected neither AT2-receptor levels nor plaque composition. CONCLUSIONS: AT2-receptor is expressed in human atherosclerotic plaque. Furthermore, we detected no functionally important role of AT2-receptor expression and found no evidence that ARB treatment regulates AT2 receptor expression. PMID- 18404605 TI - Angiotensinogen gene variants in a Pakistani hypertensive population of Punjab. AB - BACKGROUND: The renin-angiotensin-aldosterone system (RAAS) plays a key role in blood pressure (BP) regulation. Among the components of the RAAS, the gene for the angiotensinogen (AGT) has been extensively studied. Several studies in different populations link Threonine instead of methionine at position 235 (M235T) and Methinine instead of threonine at position 174 (T174M) polymorphisms with essential hypertension. We were unable to study these polymorphisms in the Punjab population of Pakistan through routine Restriction Fragment Length Polymorphism (RFLP) method. Considering the importance of this region we decided to further investigate the 300 bp region harbouring these two single nucleotide polymorphisms. METHODS: Samples were derived from a larger study group. Polymerase chain reaction amplified fragments were subjected to either RFLP or Single Strand Conformation Polymorphism. Single stranded DNA showing mobility shift on denaturing gel were sequenced. RESULTS: Sequencing confirmed the presence of M235T and T174M polymorphisms in the local population. In addition to these polymorphisms one additional base was found at an identical position in two of the samples. We found a substitution of G with C just adjacent to T174M polymorphism in all seven of our samples studied. CONCLUSIONS: We report two additional bases and one substitution in the angiotensinogen gene of Punjab population. We also suggest that SsmI can be used for the investigation of T174M polymorphism. PMID- 18404606 TI - Short-term effects of angiotensin receptor blockers on blood pressure control, and plasma inflammatory and fibrinolytic parameters in patients taking angiotensin-converting enzyme inhibitors. AB - INTRODUCTION: Angiotensin-converting enzyme (ACE) inhibitors reduce cardiovascular events in patients with established vascular disease and heart failure (HF). ACE-inhibitors have important effects on fibrinolytic balance, which may be the underlying mechanism for a reduction in cardiovascular events. Although angiotensin-receptor blockers (ARBs) offer greater tolerability than ACE inhibitors, the major ARB trials have demonstrated a lack of reduction in myocardial infarction (MI) occurrence and mortality in contrast to ACE inhibitors. In this study, we investigated the combined effects of ARBs and ACE inhibitors on fibrinolytic and inflammatory parameters in patients with uncontrolled hypertension. METHODS: Twenty-four patients with uncontrolled hypertension despite taking adequate doses of ACE-inhibitor therapy were selected. Patients were started on Candesartan 16 mg once a day. Plasma plasminogen activator inhibitor (PAI-1) antigen (Ag), tissue plasminogen activator (t-PA) Ag, thrombin-activatable fibrinolysis inhibitor (TAFI) % activity and high sensitivity C-reactive protein (hsCRP) levels, were measured during low salt intake at baseline and two weeks after therapy with an ARB. RESULTS: Addition of ARB to the regimen reduced systolic (155+/-17 vs. 139+/-13, p<0.001), and diastolic (91+/-9 vs. 81+/-8, p<0.001) blood pressures (BP). No significant changes were observed in PAI-1 Ag (66+/-51 vs. 68+/-52, p=0.9), t-PA Ag (12.6+/-5.3 vs. 13.3+/-4.7, p=0.3), TAFI % activity (119+/-30 vs. 118+/-32, p=0.9) and hsCRP (3.9+/-3.4 vs. 3.6+/-3.6, p=0.7) levels after adding an ARB. CONCLUSIONS: Combined ARB and ACE-inhibitor use provide better BP control without any detrimental effect in plasma inflammatory and fibrinolytic parameters. PMID- 18404607 TI - Lack of association between the angiotensin-converting enzyme gene (I/D) polymorphism and diabetic nephropathy in Tunisian type 2 diabetic patients. AB - OBJECTIVE: The aim of the present study was to investigate whether the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism is associated with diabetic nephropathy and type 2 diabetes in the Tunisian population. DESIGN: A case-control study was conducted among 141 unrelated type 2 diabetic patients with (90 patients) or without nephropathy (51 patients) and 103 non-diabetic controls with normal fasting blood glucose. Genotyping was performed using a nested polymerase chain reaction amplification in order to identify correctly heterozygous individuals. RESULTS: The distribution of DD, ID and II genotypes did not significantly differ between type 2 diabetic patients with or without nephropathy (DD: 44%; ID: 46%; II: 10% vs. DD: 41%; ID: 47 %; II: 12%, respectively). There was also no significant statistical difference between the genotype distribution and allele frequencies of the (I/D) polymorphism in all type 2 diabetic subjects compared to non-diabetic controls with normal fasting blood glucose (DD: 43%; ID: 46%; II: 11% vs. DD: 37%; ID: 48%; II: 15%, respectively). CONCLUSIONS: In the present preliminary study, the (I/D) polymorphism within the ACE gene is likely not associated with diabetic nephropathy nor with type 2 diabetes in the Tunisian studied population. PMID- 18404608 TI - Angiotensin-converting enzyme activity and cognitive impairment during hypoglycaemia in healthy humans. AB - INTRODUCTION: In type 1 diabetes increased risk of severe hypoglycaemia is associated with high angiotensin-converting enzyme (ACE) activity. We tested in healthy humans the hypothesis that this association is explained by the reduced ability of subjects with high ACE activity to maintain normal cognitive function during hypoglycaemia. METHODS: Sixteen healthy volunteers selected by either particularly high or low serum ACE activity were subjected to hypoglycaemia (plasma glucose 2.7 mmol/L). Cognitive function was assessed by choice reaction tests. RESULTS: Despite a similar hypoglycaemic stimulus in the two groups, only the group with high ACE activity showed significant deterioration in cognitive performance during hypoglycaemia. In the high ACE group mean reaction time (MRT) in the most complex choice reaction task was prolonged and error rate (ER) was increased in contrast to the low ACE group. The total hypoglycaemic symptom response was greater in the high ACE group than in the low ACE group (p=0.031). There were no differences in responses of counterregulatory hormones or in concentrations of substrates between the groups. CONCLUSION: Healthy humans with high ACE activity are more susceptible to cognitive dysfunction and report higher symptom scores during mild hypoglycaemia than subjects with low ACE activity. PMID- 18404609 TI - Effect of angiotensin receptor blockade on central haemodynamics in essential hypertension: results of a randomised trial. AB - OBJECTIVE: Angiotensin-converting enzyme (ACE) inhibitors have been shown to lower central augmentation index (cAI), an index of arterial wave reflection, more than beta-blockers. We tested whether this is also true for long-term treatment with an angiotensin receptor blocker (ARB). METHODS: One-hundred and fifty-six subjects with essential hypertension were randomised to treatment with either irbesartan or atenolol. cAI and central blood pressure (BP) were determined by pulse wave analysis from the radial and the carotid artery after six and after 18 months treatment. RESULTS: Peripheral and central systolic and diastolic BP were reduced to a similar extent in the two groups. cAI was reduced with irbesartan, but increased with atenolol (derived from the carotid artery: 6+/-10 vs. -4+/-12% after six months, p<0.001; -4+/-12 vs. +1+/-11% after 18 months; p=0.011). Furthermore, central to peripheral pulse pressure (PP) amplification was unaffected by treatment with irbesartan, but decreased with atenolol. CONCLUSIONS: Although treatment with irbesartan and atenolol similarly decreased peripheral and central BP, only treatment with irbesartan had beneficial effects on arterial wave reflection and preserved PP amplification. These haemodynamic effects may at least partly explain the reported differential effects of ARB versus beta-blocker treatment on cardiovascular mortality in patients with essential hypertension. PMID- 18404610 TI - The Bothrops legacy: vasoactive peptides from Brazil. PMID- 18404611 TI - Building the future of renin science. Abstracts of the Renin Academy Summit. April 16-17, 2008. Berlin, Germany. PMID- 18404614 TI - Optimizing measurement of the electrical anisotropy of muscle. AB - Skeletal muscle is electrically anisotropic, with applied high-frequency electrical current flowing more easily along than across muscle fibers. As an early step in harnessing this characteristic for clinical use, we studied approaches for maximizing the measured anisotropy by varying electrode size and applied current frequency in the tibialis anterior of 10 normal subjects. The results were compared to those from two patients with amyotrophic lateral sclerosis (ALS). Current was applied percutaneously, first parallel and then perpendicular to the major fiber direction of the muscle at frequencies ranging from 20 kHZ to 1 MHZ, using a fixed voltage-electrode length and varying the current-electrode length. The measured anisotropy was most pronounced using the longest length current electrodes and with a 125-kHZ applied frequency for the major outcome parameter phase. In addition, the two ALS patients showed very distinct anisotropic patterns. These results support the belief that, with the appropriate measurement technique, non-invasive assessment of electrical anisotropy of muscle may have useful clinical application. PMID- 18404616 TI - The more the merrier? PMID- 18404618 TI - Erratum: the tension free vaginal tape operation for women with mixed incontinence: do preoperative variables predict the outcome? PMID- 18404619 TI - Letter to the editor: conflicts of interest. PMID- 18404620 TI - EIC editorial-27:4. PMID- 18404621 TI - Adverse drug reactions reporting by physicians in Ibadan, Nigeria. AB - OBJECTIVE: To assess knowledge, extent of reporting and factors influencing reports of adverse drug reactions (ADRs) by resident doctors. STUDY DESIGN AND SETTING: Study site was the University College Hospital (UCH), Ibadan, an 850-bed tertiary care hospital in western Nigeria. It was a cross-sectional survey. Data were collected with a 46-item self-administered questionnaire from 192 clinical physicians. RESULTS: Older aged doctors, senior cadre and doctors in medical faculty were 1.5 times likely to be aware of ADR reporting guidelines. General knowledge on ADR was good while 58.3% of residents knew the burden of ADRs. Most (89.5%) had observed at least one ADR but only 32% had ever reported it. Residents in medical faculty were twice more likely to report ADRs than the surgeons [OR 2.1; 95%CI 1.1, 4.1]. The commonest factors that militate against ADR reporting were lack of knowledge that reporting forms were available (70.9%) and ignorance of reporting procedure (69%). CONCLUSION: Despite high observation and good knowledge of ADR among doctors, the rate of reporting was low. There is need for publicity of activities of the pharmacovigilance unit of the hospital. PMID- 18404622 TI - Mixed-effects Poisson regression analysis of adverse event reports: the relationship between antidepressants and suicide. AB - A new statistical methodology is developed for the analysis of spontaneous adverse event (AE) reports from post-marketing drug surveillance data. The method involves both empirical Bayes (EB) and fully Bayes estimation of rate multipliers for each drug within a class of drugs, for a particular AE, based on a mixed effects Poisson regression model. Both parametric and semiparametric models for the random-effect distribution are examined. The method is applied to data from Food and Drug Administration (FDA)'s Adverse Event Reporting System (AERS) on the relationship between antidepressants and suicide. We obtain point estimates and 95 per cent confidence (posterior) intervals for the rate multiplier for each drug (e.g. antidepressants), which can be used to determine whether a particular drug has an increased risk of association with a particular AE (e.g. suicide). Confidence (posterior) intervals that do not include 1.0 provide evidence for either significant protective or harmful associations of the drug and the adverse effect. We also examine EB, parametric Bayes, and semiparametric Bayes estimators of the rate multipliers and associated confidence (posterior) intervals. Results of our analysis of the FDA AERS data revealed that newer antidepressants are associated with lower rates of suicide adverse event reports compared with older antidepressants. We recommend improvements to the existing AERS system, which are likely to improve its public health value as an early warning system. PMID- 18404623 TI - Comparison of mass spectra of peptides in different matrices using matrix assisted laser desorption/ionization and a multi-turn time-of-flight mass spectrometer, MULTUM-IMG. AB - The mass spectra of peptides obtained with different matrices were compared using a matrix-assisted laser desorption/ionization (MALDI) ion source and a multi-turn time-of-flight (TOF) mass spectrometer, MULTUM-IMG, which has been developed at Osaka University. Two types of solid matrices, alpha-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB), and a liquid matrix made from a mixture of 3-aminoquinoline and CHCA were used. When measuring the peak signal intensity of human angiotensin II [M+H]+ from a fixed sample position, the liquid matrix produced a stable signal over 1000 laser shots, while the signal obtained with CHCA and DHB decayed after about 300 and 100 shots, respectively. Significant differences in the mass resolving power were not observed between the spectra obtained with the three matrices. Signal peak areas were measured as a function of the cycle number in a multi-turn ion trajectory, i.e., the total flight time over a millisecond time scale. For both [M+H]+ of human angiotensin II and bovine insulin, the decay of the signal peak area was the most significant with CHCA, while that measured with DHB was the smallest. The results of the mean initial ion velocity measurements suggested that the extent of metastable decomposition of the analyte ions increased in order of DHB, the liquid matrix, and CHCA, which is consistent with the difference in the decay of the signal peak area as the total flight time increased. PMID- 18404624 TI - Disaster mental health workers responding to Ground Zero: one year later. AB - The current study examined anniversary reactions in mental health disaster relief workers following traumatic exposure at the site of the World Trade Center terrorist attacks. Despite relatively low levels of symptom reporting, workers endorsed an increase in both negative mood symptoms and functional impairment at the one-year anniversary of their traumatic exposure (compared to 6 months postexposure). For those individuals who met at least partial criteria for PTSD immediately following exposure, overall self-reported PTSD symptoms tended to increase from 6 to 12 months. This tendency resulted specifically from an increase in hyperarousal symptoms. Although few endorsed symptoms at clinical levels, our results demonstrate that disaster relief workers may experience an increase in symptomatology at the anniversary of their traumatic exposure. PMID- 18404626 TI - Mutations, structural variations, and genome-wide resequencing: where to from here in our understanding of disease and evolution? AB - The 9th International Symposium on Mutations in the Genome, Mutation Detection 2007, was held on 23-27 September 2007 in Xiamen, China. Meeting participants reported on a broad range of advances in mutation detection technologies and their applications, including developments in SNP genotyping systems applicable to point-of-care diagnostic testing; and emerging views on structural variation, high-throughput sequencing and the importance of bioinformatic tools to support the growing amounts of genome variation data. This meeting report summaries the major themes presented at the meeting. PMID- 18404625 TI - A cohort study of the long-term impact of a fire disaster on the physical and mental health of adolescents. AB - The literature on adult trauma survivors demonstrates that those exposed to traumatic stress have a poorer physical health status than nonexposed individuals. Studies on physical health effects in adolescent trauma survivors, in contrast, are scarce. In the current study, it was hypothesized that adolescents who have been involved in a mass burn incident (N = 124) will demonstrate more physical and mental health problems than an unaffected cohort from the same community (N = 1,487). Health data were extracted from electronic medical records, covering 1-year prefire and 4-years postfire. When compared to the prefire baseline, survivors showed significantly larger increases in mental, respiratory, and musculoskeletal problems than community controls during the first year after the fire, but not during the later years. PMID- 18404627 TI - Accumulated childhood trauma and symptom complexity. AB - The relationship between accumulated exposure to different types of traumatic events (cumulative trauma) in childhood and the total number of different types of symptomatology reported (symptom complexity) in adulthood was examined in a sample of 2,453 female university students. There was a linear relationship between the number of trauma types experienced by participants before 18 and symptom complexity. This effect remained even when controlling for specific traumatic events, suggesting a generalized effect of cumulative trauma. PMID- 18404628 TI - Unpacking the relationship between posttraumatic numbing and hyperarousal in a sample of help-seeking motor vehicle accident survivors: replication and extension. AB - The current study is a replication and extension of previous research that has found support for a relationship between posttraumatic numbing and hyperarousal. This study examined this association while controlling for depression in 345 motor vehicle accident survivors. Additionally, the relationships among specific hyperarousal symptoms and numbing were explored. Results provided further evidence for an association between hyperarousal and numbing, even while controlling for the influence of depression, and revealed that all hyperarousal symptoms (except hypervigilance) contribute to this association. PMID- 18404629 TI - Elevated rates of current PTSD among Hispanic veterans in the NVVRS: true prevalence or methodological artifact? AB - The elevated rate of current posttraumatic stress disorder (PTSD) among Hispanic Vietnam veterans has been attributed to culturally based expressiveness that inflates symptom self-reports. To investigate this possibility, the authors conducted three hypothesis-driven analyses with National Vietnam Veterans Readjustment Study (NVVRS) data from the Structured Clinical Interview for DSM III-R (SCID-) diagnosed subsample of male Vietnam Theater veterans (N = 260). First, persistence of the Hispanic elevation after adjusting for war-zone stress exposure initially suggested the effect of greater expressiveness. Second, symptom-based analyses isolated this effect to the self-report Mississippi Scale for Combat-Related PTSD and not to the clinician-rated SCID interview. Third, objective measures of functioning did not reveal a unique Hispanic pattern of lower impairment associated with current PTSD. These tests suggest that greater Hispanic expressiveness does not account for the Hispanic elevation in current PTSD in the NVVRS SCID-diagnosed subsample. PMID- 18404630 TI - War-related posttraumatic stress disorder in Black, Hispanic, and majority White Vietnam veterans: the roles of exposure and vulnerability. AB - Elevated prevalence rates of chronic posttraumatic stress disorder (PTSD) have been reported for Black and Hispanic Vietnam veterans. There has been no comprehensive explanation of these group differences. Moreover, previous research has relied on retrospective reports of war-zone stress and on PTSD assessments that fail to distinguish between prevalence and incidence. These limitations are addressed by use of record-based exposure measures and clinical diagnoses of a subsample of veterans from the National Vietnam Veterans Readjustment Study (NVVRS). Compared with Majority White, the Black elevation is explained by Blacks' greater exposure; the Hispanic elevation, by Hispanics' greater exposure, younger age, lesser education, and lower Armed Forces Qualification Test scores. The PTSD elevation in Hispanics versus Blacks is accounted for mainly by Hispanics' younger age. PMID- 18404631 TI - The factor structure of the posttraumatic growth inventory: a comparison of five models using confirmatory factor analysis. AB - There are different views about the dimensions of the positive changes resulting from the struggle with traumatic events. Using Posttraumatic Growth Inventory (PTGI) data reported by participants (N = 926) experiencing a variety of traumatic events, five models of the underlying structure of the PTGI were tested via confirmatory factor analyses to examine whether the PTGI comprises three domains (Changed Perception of Self, Changed Interpersonal Relationships, and Changed Philosophy of Life), five factors (Relating to Others, New Possibilities, Personal Strength, Spiritual Change, and Appreciation of Life), or a unitary dimension. Results indicated an oblique 5-factor model best fit the data, thus revealing the PTGI was multidimensional. Present findings offer implications for understanding the nature of posttraumatic growth. PMID- 18404632 TI - Strengths of character and posttraumatic growth. AB - How are strengths of character related to growth following trauma? A retrospective Web-based study of 1,739 adults found small, but positive associations among the number of potentially traumatic events experienced and a number of cognitive and interpersonal character strengths. It was concluded that growth following trauma may entail the strengthening of character. PMID- 18404633 TI - Symptom comparison across multiple solicitation methods among Burundians with traumatic event histories. AB - Debate continues over whether posttraumatic stress disorder (PTSD) symptoms are more biologically based and therefore relatively universal or are more culturally constructed. This study aimed to describe traumatic stress reactions in a Burundian sample and to investigate the influences of the solicitation method (open-ended questions and standardized measures) and psychoeducation (as a process of acculturation) on symptoms reported. Standardized measures showed that distress was manifested in somatization, anxiety, and depression, and less so in specific PTSD symptoms. Content analysis of open-ended questions revealed frequent material complaints. Prior exposure to Western ideas about trauma was predictive of more severe PTSD symptoms. The implications of the findings are discussed in terms of how methodological and cultural factors may influence posttraumatic reactions in nonindustrialized settings. PMID- 18404634 TI - A field test of group based exposure therapy with 102 veterans with war-related posttraumatic stress disorder. AB - Group-based exposure therapy (GBET) was field-tested with 102 veterans with war related posttraumatic stress disorder (PTSD). Nine to 11 patients attended 3 hours of group therapy per day twice weekly for 16-18 weeks. Stress management and a minimum of 60 hours of exposure was included (3 hours of within-group war trauma presentations per patient, 30 hours of listening to recordings of patient's own war-trauma presentations and 27 hours of hearing other patients' war-trauma presentations). Analysis of assessments conducted by treating clinicians pre-, post- and 6-month posttreatment suggests that GBET produced clinically significant and lasting reductions in PTSD symptoms for most patients on both clinician symptoms ratings (6-month posttreatment effect size delta = 1.22) and self-report measures with only three dropouts. PMID- 18404635 TI - Acute dissociation predicts rapid habituation of skin conductance responses to aversive auditory probes. AB - The present study examined how acute dissociation, trait-like dissociative symptoms, and physiological reactivity relate to each other. Sixty-nine undergraduate students were exposed to 14 aversive auditory probes, while their skin conductance responses were measured. A combination of self-reported anxiety and trait-like dissociation was found to predict variability in peritraumatic dissociation levels induced by the aversive probes. Furthermore, high levels of acute dissociation were associated with faster habituation of skin conductance responding, while trait-like dissociation was unrelated to habituation. Interestingly, individuals who reported childhood trauma displayed elevated skin conductance responses. Our findings contribute to the growing body of evidence indicating that subjective feelings of acute dissociation have their objective concomitants, notably fast habituation of physiologic responses. PMID- 18404636 TI - PTSD diagnoses, subsyndromal symptoms, and comorbidities contribute to impairments for breast cancer survivors. AB - The clinical importance of posttraumatic stress disorder (PTSD) symptomatology for cancer patients is unclear. The association between the magnitude of cancer related PTSD symptoms, comorbidity, and functioning is tested. Breast cancer patients (N = 74) were assessed at diagnosis/surgery, followed, and screened for cancer-related PTSD 18 months later. Participants then completed diagnostic interviews and PTSD (n = 12), subsyndromal PTSD (n = 5), and no symptom (n = 47) patient groups were identified. Posttraumatic stress disorder cases were distinguished by having experienced violent traumas and anxiety disorders predating cancer, whereas subsyndromal cases were not. Also, longitudinal data show that PTSD covarys with poorer functioning and lower quality of life among breast cancer survivors. Both PTSD and subsyndromal PTSD were associated with employment absenteeism and the seeking of mental health services. PMID- 18404638 TI - Association of child sexual abuse and dating victimization with mental health disorder in a sample of adolescent girls. AB - The study's aim was to assess the association between adolescent girls' recall of child sexual abuse (CSA) and prevalence of dating victimization in early romantic relationships and to explore the link between multiple trauma history and mental disorders. When demographics and teacher-reported behavioral problems during childhood were controlled, the risk of presenting with at least one internalized or externalized mental health disorder was higher for adolescent girls with multiple victimization (history of CSA and dating victimization) than for adolescents with no multiple trauma history. Results highlight a strong association between reports of multiple traumatic experiences and mental health disorders of teenaged girls and the need to assess for a broader range of victimization experiences in the design of intervention services. PMID- 18404637 TI - PTSD symptom clusters are differentially related to substance use among community women exposed to intimate partner violence. AB - Women who experience intimate partner violence (IPV) have higher rates of posttraumatic stress disorder (PTSD) and substance abuse compared to women who do not experience IPV. However, the extent to which IPV-related PTSD symptoms are related to women's substance use involvement largely has been unexplored. The current study investigated PTSD symptomatology and substance use in a community sample of 212 IPV-exposed women. Drug-using women reported higher PTSD severity scores compared to women who reported no substance use or alcohol use only. Moreover, the reexperiencing, avoidance and numbing, and arousal clusters demonstrated unique associations with substance use involvement. Findings not only elucidate the associations among IPV-related PTSD symptoms and substance use, but they also can inform community-based preventive interventions. PMID- 18404639 TI - Mechanisms of anger and treatment outcome in combat veterans with posttraumatic stress disorder. AB - Research has identified anger as prominent in, and an influence on, treatment outcome for military veterans with posttraumatic stress disorder (PTSD). This study examined factors influencing the relationship between anger and outcome to improve treatment effectiveness. Participants comprised 103 veterans attending PTSD treatment. Measures of PTSD and comorbidity were obtained at intake and 9 month follow-up. Measures also included potential mediators of therapeutic alliance, social support, problematic/undermining relationships and fear of emotion. Path analyses supported anger as a predictor of treatment outcome, with only fear of anger and alcohol comorbidity accounting for the variance between anger and outcome. To improve treatment effectiveness, clinicians need to assess veterans' anger, aggression, and alcohol use, as well as their current fear of anger and elucidate the relationship between these factors. PMID- 18404640 TI - Resting electroencephalogram asymmetry and posttraumatic stress disorder. AB - The valence-arousal (W. Heller, 1993) and approach-withdrawal (R. J. Davidson, 1998a) models hypothesize that particular patterns of hemispheric brain activity are associated with specific motivational tendencies and psychopathologies. We tested several of these predictions in two groups-a posttraumatic stress disorder (PTSD) and a "supercontrol" group, selected to be maximally different from those with PTSD. Contrary to almost all hypotheses, individuals with PTSD did not differ from controls on resting electroencephalogram (EEG) asymmetry. Particular aspects of PTSD were also not related to EEG hemisphere differences. Our null findings are consistent with the few studies that have examined resting EEG asymmetries in PTSD and suggest that PTSD may be associated with different processes than psychopathologies previously examined in studies of hemispheric brain activity (e.g., major depressive disorder, panic disorder). PMID- 18404641 TI - Entering exile: trauma, mental health, and coping among Tibetan refugees arriving in Dharamsala, India. AB - Each year thousands of Tibetans escape Chinese-controlled Tibet. The authors present findings on the experiences, coping strategies, and psychological distress (depression, anxiety, somatization, and posttraumatic stress disorder) of 769 Tibetan refugees arriving in Dharamsala, India (2003-2004). Distress increased significantly with greater trauma exposure. However, despite a high prevalence of potentially traumatizing events, levels of psychological distress were extremely low. Coping activity (primarily religious) and subjective appraisals of trauma severity appeared to mediate the psychological effects of trauma exposure. The potential impact of other variables, including culturally determined attitudes about trauma and timing of assessment, are discussed. PMID- 18404642 TI - Anorexia nervosa and parental bonding: the contribution of parent-grandparent relationships to eating disorder psychopathology. AB - The present study adopted an intergenerational approach in examining the association between parental bonding and anorexia nervosa. Forty-three anorexic participants and 33 nonclinical comparison participants completed eating disorder questionnaires and the Parental Bonding Instrument (PBI). The participant's parents also completed the PBI. The anorexic participants perceived both parents as less caring and fathers as more controlling than nonclinical participants. Among anorexic participants, mother control and father care were associated with symptom severity. Intergenerational effects were present. Among anorexic participants, maternal grandmother care was associated with eating disorder psychopathology. The present findings suggest that parental characteristics of grandparents might play a role in the development of eating disorders in granddaughters. PMID- 18404643 TI - Juvenile and adult immature and in vitro matured ovine oocytes evaluated in relation to membrane electrical properties, calcium stores, IP3 sensitivity and apoptosis occurrence in cumulus cells. AB - The analysis of differences between juvenile and adult oocytes may provide useful information on the acquisition of meiotic and developmental competence of the female gamete. In oocytes collected from either ewes or 40-day-old lambs, we evaluated membrane electrical properties, such as resting potential, conductance, activation ion currents, L-type Ca(2+) currents as well as calcium stores and IP3 sensitivity; in addition, the incidence of apoptosis in cumulus cells in these two age categories was compared. The analysis was carried out in oocytes both prior to and after in vitro maturation. Significant differences were found in all the examined parameters in relation to maturational stages whereas minor differences were recorded in relation to age of the donor. IP3 sensitivity strongly increased after in vitro maturation following a dose-dependent pattern from 1 to 500 micromol/L with a significant interaction (P < 0.01) between dose and maturational stage. The incidence of apoptosis in cumulus cells strongly increased after in vitro maturation and was greater in adult than in juvenile cumulus cells (39.2 +/- 5.8% vs. 21.9 +/- 3.5%; P < 0.01). In conclusion, all the examined parameters were greatly affected by the maturational stage, whereas minor differences were due to age-related oocyte quality, that is, at plasma membrane levels to conductance, activation current peaks and calcium currents, at cytosol level to calcium stores and IP3 sensitivity, and to incidence of apoptosis in cumulus cells. These parameters were compared with previous data in bovine to analyze oocyte quality in juvenile and adult individuals or between species. PMID- 18404644 TI - Genetic association between alpha-synuclein and idiopathic Parkinson's disease. AB - Point mutations and copy number variations in SNCA, the gene encoding alpha synuclein, cause familial Parkinson's disease (PD). A dinucleotide polymorphism (REP1) in the SNCA promoter may be a risk factor for common forms of PD. We studied 1,802 PD patients and 2,129 controls from the NeuroGenetics Research Consortium, using uniform, standardized protocols for diagnosis, subject recruitment, data collection, genotyping, and data analysis. Three common REP1 alleles (257, 259, and 261 bp, with control frequencies of 0.28, 0.65, and 0.06) and several rare alleles (combined frequency <0.01) were detected. We confirmed association of REP1 with PD risk [odds ratio (OR) = 0.86, P = 0.006 for 257 carriers; OR = 1.25, P = 0.022 for 261-carriers]. Using a normalization procedure, we showed that the 257 and 261 alleles are both independently associated with PD risk (for 257, P = 0.002 in overall data, 0.003 in non familial PD, 0.001 in early-onset PD; for 261, P = 0.056 in overall data, 0.024 in non-familial PD, 0.052 in early-onset PD). The 257-associated risk was consistent with a dominant model [hazard ratio (HR) = 0.99, P = 0.91 for 257/257 vs. 257/X where X denotes all other common alleles; HR = 1.16, P = 0.004 for X/X vs. 257/X]. The 261-associated risk was consistent with a recessive model (HR = 1.89, P = 0.026 for 261/261 vs. 261/X; HR = 0.95, P = 0.42 for X/X vs. 261/X). Genotype-specific mean onset ages (+/-SD) ranged from 54.8 +/- 12.1 for 261/261 to 59.4 +/- 11.5 for 257/257, displaying a trend of decreasing onset age with increasing allele size (P = 0.055). Genetic variation in SNCA and its regulatory regions play an important role in both familial and sporadic PD. PMID- 18404646 TI - Geminin is overexpressed in human pancreatic cancer and downregulated by the bioflavanoid apigenin in pancreatic cancer cell lines. AB - Pancreatic adeniocarcinoma is among the deadliest of human cancers. Apigenin, an antitumor flavonoid, inhibits pancreatic cancer cell proliferation in vitro. Geminin is a recently identified novel protein that plays a critical role in preventing abnormal DNA replication by binding to and inhibiting the essential replication factor Cdt1. Microarray analysis identified geminin to be downregulated in pancreatic cancer cells treated with apigenin. Therefore, we investigated the effects of apigenin on geminin expression and other proteins involved in replication (Cdc6, Cdt1, and MCM7) in pancreatic cancer cell lines CD18 and S2013. Real time RT-PCR and western blotting analysis showed that geminin expression is downregulated by apigenin at both mRNA and protein levels. Furthermore, treatment of cells with proteosome inhibitor MG132 reversed the downregulation of geminin by apigenin, supporting our hypothesis that the degradation pathway is another mechanism by which apigenin affects geminin expression. Apigenin treatment also resulted in downregulation of Cdc6 at both mRNA and protein levels. However, Cdt1 and MCM7 expression was not affected in apigenin-treated cells. The effect of apigenin treatment on geminin promoter activity was measured by transient transfection of Hela cells with a reporter gene, demonstrating that apigenin inhibited geminin promoter activity. Geminin expression was also evaluated in human pancreatic tissue (n = 15) by immunohistochemistry and showed that geminin is overexpressed in human pancreatic cancer compared to normal adjacent pancreatic tissue. In conclusion, our studies demonstrated that geminin is overexpressed in human pancreatic cancer and downregulated by apigenin which may contribute to the antitumor effect of this natural flavonoid. PMID- 18404645 TI - FBXL21 association with schizophrenia in Irish family and case-control samples. AB - FBXL21 gene encodes an F-box containing protein functioning in the SCF ubiquitin ligase complex. The role of the F-box protein is to recruit proteins designated for degradation to the ligase complex so they would be ubiquitinated. Using both family and case-control samples, we found consistent associations in and around FBXL21 gene. In the family sample (Irish study of high density schizophrenia families, ISHDSF, 1,350 subjects from 273 families), a minimal PDT P-value of 0.0011 was observed at rs31555. In the case-control sample (Irish case-control study of schizophrenia, ICCSS, 814 cases and 625 controls), significant associations were observed at two markers (rs1859427 P = 0.0197, and rs6861170 P = 0.0197). In haplotype analyses, haplotype 1-1 (C-T) of rs1859427-rs6861170 was overtransmitted in the ISHDSF (P = 0.0437) and was overrepresented in the ICCSS (P = 0.0177). For both samples, the associated alleles and haplotypes were identical. These data suggested that FBXL21 may be associated with schizophrenia in the Irish samples. PMID- 18404647 TI - Meta-analysis of alexithymia in posttraumatic stress disorder. AB - The authors present a meta-analysis investigating the prevalence of alexithymia in 12 studies encompassing 1,095 individuals with posttraumatic stress disorder (PTSD). A large effect size was found associating PTSD with alexithymia. Effect sizes were higher in studies of male combat PTSD samples in comparison with studies of other PTSD samples. Clinical and research directions are discussed. PMID- 18404648 TI - Virtual reality exposure therapy using a virtual Iraq: case report. AB - Posttraumatic stress disorder (PTSD) has been estimated to affect up to 18% of returning Operation Iraqi Freedom (OIF) veterans. Soldiers need to maintain constant vigilance to deal with unpredictable threats, and an unprecedented number of soldiers are surviving serious wounds. These risk factors are significant for development of PTSD; therefore, early and efficient intervention options must be identified and presented in a form acceptable to military personnel. This case report presents the results of treatment utilizing virtual reality exposure (VRE) therapy (virtual Iraq) to treat an OIF veteran with PTSD. Following brief VRE treatment, the veteran demonstrated improvement in PTSD symptoms as indicated by clinically and statistically significant changes in scores on the Clinician Administered PTSD Scale (CAPS; Blake et al., 1990) and the PTSD Symptom Scale Self-Report (PSS-SR; Foa, Riggs, Dancu, & Rothbaum, 1993). These results indicate preliminary promise for this treatment. PMID- 18404649 TI - Treatment barriers for low-income, urban African Americans with undiagnosed posttraumatic stress disorder. AB - African Americans in low-income, urban communities are at high risk for exposure to traumatic events as well as for symptoms of posttraumatic stress disorder (PTSD). Approximately 22% of 220 participants recruited from urban hospital medical clinics met survey criteria for PTSD. Among the common traumas were having relatives/friends murdered (47%), being attacked with weapons (64% of men), and being sexually attacked (36% of women). Although desiring mental health services, only 13.3% of those with PTSD had prior trauma-focused treatment. Barriers to treatment included limited transportation and finances, family disapproval, and unfamiliarity with accessing treatment, among others. These data highlight the need for an awareness of the high prevalence of trauma and PTSD in this population. PMID- 18404650 TI - PATH's approach to product introduction in developing countries. AB - Health interventions developed for wealthy nations are difficult to introduce in developing countries due to a lack of sufficient technological, financial, political, or infrastructural resources. Increasingly, however, product developers are creating new technologies that meet the needs of low-resource settings. Experience has shown that many activities must be undertaken to prepare health systems to accept and embrace new, improved, or under-utilized health technologies. This article describes three health technologies--a diagnostic test for vitamin A deficiency, new cervical cancer vaccines, and the vaccine vial monitor--that PATH, a global health organization, has helped advance for developing-country markets. PMID- 18404651 TI - Intervertebral disc degeneration in a naturally occurring primate model: radiographic and biomechanical evidence. AB - Classic degenerative disc disease is a serious health problem worldwide, whose etiological basis-mechanical stimulus, biochemical changes, or natural aging-is poorly understood. Animal models are critical to the study of degenerative disc disease initiation and progression and for attempts to regulate, ameliorate, or eliminate it. The macaque represents a primate model with natural disc degeneration that might serve to advance the field; we aimed to provide radiographic (morphologic) and biomechanical evidence of natural disc degeneration in this model. A factorial study design was used to examine the relationship between the radiographic appearance of disc degeneration and its biomechanical consequences. Eighteen macaques of advanced age (22.3 +/- 0.9 years) had radiographs taken to assess the degree of thoracolumbar intervertebral disc degeneration using a standard atlas method. Each spine was harvested and dynamic biomechanical tests were performed. Advancing disc degeneration (degree of disc space narrowing and osteophytosis) was associated with increased stiffness, decreased energy absorption, and increased natural frequency of the intervertebral disc. These associations linking the dynamics of the intervertebral disc and its degree of degeneration are similar to those found in humans. Our results indicate the macaque model with morphologic and biomechanical efficacy could aid in understanding the progression of disc degeneration and in developing therapeutic strategies to prevent or inhibit its course. PMID- 18404652 TI - Gene expression profiles of human chondrocytes during passaged monolayer cultivation. AB - Chondrocyte phenotype has been shown to dedifferentiate during passaged monolayer cultivation. Hence, we have investigated the expression profile of 27 chondrocyte associated genes from both osteoarthritic cartilage tissue and healthy passaged human articular chondrocytes by quantitative real-time PCR. Our results indicate that the gene expression levels of matrix proteins and proteases in chondrocytes from monolayer culture decrease compared with those from cartilage tissue, while monolayer cultured chondrocytes from normal and osteoarthritic cartilage exhibit similar gene expression patterns. However, chondrocytic gene expression profiles were differentially altered at various stages of passage. The expression of the matrix proteins aggrecan, type II collagen, and fibromodulin inversely correlated with increasing passage number, while fibronectin and link protein exhibited a marked increase with passage. The expression of matrix proteinases MMP-3/9/13 and ADAMTS-4/5 decreased with passage, whereas proteinase inhibitors TIMP-2/3 were elevated. The cytokine IL-1 also showed increased expression with monolayer chondrocyte culture, while IGF-1 expression levels were diminished. No significant changes in TGF-beta, or the chondrogenic transcription factors Sox-9, c-fos, or c-jun were observed. Our data indicates that cultured chondrocytes undergo dedifferentiation during monolayer culture, although the gene expression level of transcription factors necessary for chondrogenesis remains unchanged. This data may prove important for the future development of more specific and efficacious cultivation techniques for human articular chondrocyte-based therapies. PMID- 18404653 TI - The influence of wear paths produced by hip replacement patients during normal walking on wear rates. AB - Variation in wear paths is known to greatly affect wear rates in vitro, with multidirectional paths producing much greater wear than unidirectional paths. This study investigated the relationship between multidirectional motion at the hip joint, as measured by aspect ratio, sliding distance, and wear rate for 164 hip replacements. Kinematic input from three-dimensional gait analysis was used to determine the wear paths. Activity cycles were determined for a subgroup of 100 patients using a pedometer study, and the relationship between annual sliding distance and wear rate was analyzed. Poor correlations were found between both aspect ratio and sliding distance and wear rate for the larger group and between annual sliding distance and wear rate for the subgroup. However, patients who experienced a wear rate <0.08 mm/year showed a strong positive correlation between the combination of sliding distance, activity levels, and aspect ratio and wear rate (adjusted r(2) = 55.4%). This group may represent those patients who experience conditions that most closely match those that prevail in simulator and laboratory tests. Although the shape of wear paths, their sliding distance, and the number of articulation cycles at the hip joint affect wear rates in simulator studies, this relationship was not seen in this clinical study. Other factors such as lubrication, loading conditions and roughness of the femoral head may influence the wear rate. PMID- 18404654 TI - Collagen orientation in periosteum and perichondrium is aligned with preferential directions of tissue growth. AB - A feedback mechanism between different tissues in a growing bone is thought to determine the bone's morphogenesis. Cartilage growth strains the surrounding tissues, eliciting alterations of its matrix, which in turn, creates anisotropic stresses, guiding directionality of cartilage growth. The purpose of this study was to evaluate this hypothesis by determining whether collagen fiber directions in the perichondrium and periosteum align with the preferential directions of long bone growth. Tibiotarsi from chicken embryos across developmental stages were scanned using optical projection tomography (OPT) to assess preferential directions of growth at characteristic sites in perichondrium and periosteum. Quantified morphometric data were compared with two-photon laser-scanning microscopy images of the three-dimensional collagen network in these fibrous tissues. The diaphyseal periosteum contained longitudinally oriented collagen fibers that aligned with the preferential growth direction. Longitudinal growth at both metaphyses was twice the circumferential growth. This concurred with well developed circumferential fibers, which covered and were partly interwoven with a dominant network of longitudinally oriented fibers in the outer layer of the perichondrium/periosteum at the metaphysis. Toward both articulations, the collagen network of the epiphyseal surface was randomly oriented, and growth was approximately biaxial. These findings support the hypothesis that the anisotropic architecture of the collagen network, detected in periosteum and perichondrium, concurs with the assessed growth directions. PMID- 18404655 TI - Correlation of dynamic cartilage contact stress aberrations with severity of instability in ankle incongruity. AB - Joint instability is presumed to cause abnormality in cartilage contact mechanics, which accumulatively damages the articular surface, leading to osteoarthritis. The purpose of this study was to clarify the effect of instability on dynamic cartilage contact mechanics. Using human ankle cadaver specimens, potentially unstable ankles were modeled by introducing a coronally directed step-off incongruity of the anterior tibial surface and/or by transecting the anterior talofibular ligament. Specimens were subjected to a duty cycle with quasi-physiologic stance-phase motion and loading. AP tibial forces were modulated, causing a controlled, quantifiable ankle subluxation during the duty cycle. Instantaneous changes in local articular contact stresses were continuously measured using a thin, flexible pressure transducer. Tests were repeated while varying the tibial surface condition (anatomic, 1-mm step-off, and 2-mm step-off), both before and after transection of the anterior talofibular ligament, with various AP force magnitudes, so that situations of various degrees of instability were created for each specimen. Instability events occurred when the step-off incongruity was introduced, with the abnormality in joint kinematics being greater after ligament transection. Contact stress data revealed that these instability events involved distinctly abrupt increases/decreases in local articular contact stresses, and that the degree of abruptness was correlated nearly linearly with the abnormality in kinematics. The severity of contact stress aberration appeared to be correlated with the degree of instability. Given this linear relationship, even small instability events presumably involve appreciable abnormality in dynamic joint contact mechanics. PMID- 18404656 TI - Pulsed electromagnetic fields enhance BMP-2 dependent osteoblastic differentiation of human mesenchymal stem cells. AB - Mesenchymal stem cells (MSCs) express an osteoblastic phenotype when treated with BMP-2, and BMP-2 is used clinically to induce bone formation although high doses are required. Pulsed electromagnetic fields (PEMF) also promote osteogenesis in vivo, in part through direct action on osteoblasts. We tested the hypothesis that PEMF enhances osteogenesis of MSCs in the presence of an inductive stimulus like BMP-2. Confluent cultures of human MSCs were grown on calcium phosphate disks and were treated with osteogenic media (OM), OM containing 40 ng/mL rhBMP-2, OM + PEMF (8 h/day), or OM + BMP-2 + PEMF. MSCs demonstrated minor increases in alkaline phosphatase (ALP) during 24 days in culture and no change in osteocalcin. OM increased ALP and osteocalcin by day 6, but PEMF had no additional effect at any time. BMP-2 was stimulatory over OM, and PEMF + BMP-2 synergistically increased ALP and osteocalcin. PEMF also enhanced the effects of BMP-2 on PGE2, latent and active TGF-beta1, and osteoprotegerin. Effects of PEMF on BMP-2-treated cells were greatest at days 12 to 20. These results demonstrate that PEMF enhances osteogenic effects of BMP-2 on MSCs cultured on calcium phosphate substrates, suggesting that PEMF will improve MSC response to BMP-2 in vivo in a bone environment. PMID- 18404657 TI - Muscle stabilization strategies in people with medial knee osteoarthritis: the effect of instability. AB - The sensation of knee instability (shifting, buckling. and giving way) is common in people with medial knee osteoarthritis (OA). Its influence on knee stabilization strategies is unknown. This study investigated the influence of knee instability on muscle activation during walking when knee stability was challenged. Twenty people with medial knee OA participated and were grouped as OA Stable (OAS) (n = 10) and OA Unstable (OAU) (n = 10) based on self-reported knee instability during daily activities. Quadriceps strength, passive knee laxity, and varus alignment were assessed and related to knee instability and muscle cocontraction during walking when the support surface translated laterally. Few differences in knee joint kinematics between the groups were seen; however, there were pronounced differences in muscle activation. The OAU group used greater medial muscle cocontraction before, during, and following the lateral translation. Self-reported knee instability predicted medial muscle cocontraction, but medial laxity and limb alignment did not. The higher muscle cocontraction used by the OAU subjects appears to be an ineffective strategy to stabilize the knee. Instability and high cocontraction can be detrimental to joint integrity, and should be the focus of future research. PMID- 18404659 TI - Manipulating the anabolic and catabolic response in bone graft remodeling: synergism by a combination of local BMP-7 and a single systemic dosis of zoledronate. AB - Remodeling of a bone graft can be influenced both by anabolic substances, such as a bone morphogenic protein (BMP) and by anticatabolic substances, such as the bisphosphonates. BMPs are potent bone anabolic substances, but also boost catabolism and cause resorption. Bisphosphonates inhibit osteoclast function and can be used to postpone resorption. In the present study a combination of both drugs was explored in a rat bone chamber model. Cancellous bone grafts were treated with either BMP-7 or saline and placed in a bone chamber implanted in the proximal tibia. After 2 weeks, an injection of either zoledronate 0.1 mg/kg or saline was given subcutaneously. The rats were killed after 6 weeks, and bone ingrowth distance into the graft and graft resorption were measured by histomorphometry. BMP-7 significantly (p = 0.007) increased new bone ingrowth distance into the graft from 2.0 mm (SD = 0.98 mm) in the controls to 3.1 mm (SD = 0.93 mm). If bisphosphonate was not given, most of the newly formed and old graft bone was resorbed. A single injection of zoledronate significantly (p < 0.001) increased the trabecular volume/total volume to 40% (SD = 9%) compared to 14% (SD = 10%) in the nonbisphosphonate treated. In total, the net amount of bone increased by 400% when BMP-7 and zoledronate combined was compared to saline. A bone graft can be treated with BMP-7 to increase new bone formation and at the same time be protected against premature catabolism by a single dose of a bisphosphonate. This combination might be useful in various conditions in orthopedic reconstruction. PMID- 18404658 TI - The effect of hyaluronidase, phospholipase, lipid solvent and trypsin on the lubrication of canine flexor digitorum profundus tendon. AB - Although the flexor digitorum profundus (FDP) tendon's gliding resistance is low, the lubrication mechanism that enables this is unclear. The principal lubricants in joints, such as hyaluronic acid, phospholipids, and lubricin, a lubricating glycoprotein, are known to be present in the canine tendon. In this study, we assessed the effect of these lubricants in the tendon by measuring gliding resistance before and after their removal. Canine FDP tendons were treated with hyaluronidase, phospholipase, lipid solvent, and/or trypsin. The gliding resistance of FDP tendons significantly increased after all treatments (p < 0.05). The largest effect on gliding resistance was observed after trypsin digestion. Scanning electron microscopy and immunostaining for hyaluronic acid and lubricin were used to qualitatively assess the tendon surface after treatments. The trypsin digestion produced the most irregular surface, with many exposed collagen fibers. The results of this study suggest that phospholipids, hyaluronic acid, and protein components are all involved in maintaining the low gliding resistance of the FDP tendon. PMID- 18404660 TI - Proprioception assessment in subjects with idiopathic loss of shoulder range of motion: joint position sense and a novel proprioceptive feedback index. AB - We examined the effects of elevation range and plane on shoulder joint proprioception in subjects with idiopathic loss of shoulder range of motion (ROM). Joint position sense (JPS) and a novel proprioceptive feedback index (PFI), including difference magnitude and the similarity index, were used to assess proprioception. Twelve subjects (eight male, four female) with involved stiff shoulders and normal opposite shoulders were recruited from a university hospital. Subjects attempted to repeat six target positions. Target positions consisted of arm elevation in three planes (frontal, scapular, and sagittal planes) and two ranges (end/mid range). Six trials of each target position were used to determine acceptable trials for stabilization of the data, less than 5% of the cumulative mean values for at least three successive trials. The data stabilized at the sixth repetition. Compared to control shoulders, involved shoulders had enhanced proprioception during end range movements (p < 0.05). The magnitude of the repositioning error and difference magnitude decreased (1.6 degrees -3.5 degrees for repositioning error and 22.2 degrees -62.1 degrees for difference magnitude), whereas similarity index improved at end range movements compared to mid range movements (p < 0.05) in involved stiff shoulders. Results of JPS and PFI suggest that both capsuloligamentous and musculotendinous mechanoreceptors play an important role in proprioception feedback during active movements in subjects with idiopathic loss of shoulder ROM. PMID- 18404661 TI - Changes of selenium, copper, and zinc content in hair and serum of patients with idiopathic scoliosis. AB - Our work aimed at extending the search for abnormalities of trace elements in patients with idiopathic scoliosis to the content of zinc (Zn), copper (Cu), and selenium (Se) in these subject's hair and serum. A total of 59 patients (54 girls and 4 boys) with idiopathic scoliosis, aged 13 on the average (range, 10-18 years), were examined. The degree of spine curvature deformity ranged between 12 degrees and 82 degrees . The hair of scoliotic patients under examination showed significantly increased content of zinc 263 +/- 108 microg/g (p < 0.01) and copper 46.2 +/- 37.1 microg/g (p < 0.01), and decreased content of selenium 0.194 +/- 0.114 microg/g (p < 0.01) in comparison with the control group. In scoliotic patients, the Cu/Zn ratio in hair (0.186 +/- 0.139) did not differ significantly from the values found in the probands of the control group (0.115 +/- 0.09). The Cu/Se ratio in this group of patients (254.9 +/- 215.9) was significantly higher (p < 0.001) due to a higher Cu value and a lower Se value in comparison with the controls (47.9 +/- 23.7). In comparison with controls, the serum selenium concentration in the group of scoliotic patients was significantly decreased p < 0.05 (0.74 +/- 0.13 micromol/L and 0.98 +/- 0.12 micromol/L). Various changes in the content of trace elements in biological samples taken from patients with idiopathic scoliosis are not accidental. What might bring about a shift in our knowledge is speciation of various forms of trace elements in the organism in relation to idiopathic scoliosis. PMID- 18404662 TI - Patient-specific finite element analysis of chronic contact stress exposure after intraarticular fracture of the tibial plafond. AB - The role of altered contact mechanics in the pathogenesis of posttraumatic osteoarthritis (PTOA) following intraarticular fracture remains poorly understood. One proposed etiology is that residual incongruities lead to altered joint contact stresses that, over time, predispose to PTOA. Prevailing joint contact stresses following surgical fracture reduction were quantified in this study using patient-specific contact finite element (FE) analysis. FE models were created for 11 ankle pairs from tibial plafond fracture patients. Both (reduced) fractured ankles and their intact contralaterals were modeled. A sequence of 13 loading instances was used to simulate the stance phase of gait. Contact stresses were summed across loadings in the simulation, weighted by resident time in the gait cycle. This chronic exposure measure, a metric of degeneration propensity, was then compared between intact and fractured ankle pairs. Intact ankles had lower peak contact stress exposures that were more uniform and centrally located. The series-average peak contact stress elevation for fractured ankles was 38% (p = 0.0015; peak elevation was 82%). Fractured ankles had less area with low contact stress exposure than intact ankles and a greater area with high exposure. Chronic contact stress overexposures (stresses exceeding a damage threshold) ranged from near zero to a high of 18 times the matched intact value. The patient specific FE models represent substantial progress toward elucidating the relationship between altered contact stresses and the outcome of patients treated for intraarticular fractures. PMID- 18404663 TI - Estimating differences between male and female physician service provision using panel data. AB - Using panel data, we estimate the impact of an increasing share of female physicians on the total output of Canadian physicians. A micro-econometric model is developed specifically for the Canadian context and estimated using administrative data on all Canadian physicians paid on a fee-for-service basis from 1989 to 1998. Our results suggest that female physicians systematically provide fewer services than their male counterparts for almost all specialties and provinces studied. Given that females account for an increasing share of the physician population and that female physicians provide, on average, fewer services, potentially important future reductions in total health-care service provision are likely. PMID- 18404664 TI - Impact of the recall period on measuring health utilities for acute events. AB - The impact of healthcare interventions on health utility values is most frequently measured using a preference-based instrument. Each of the available instruments instructs the respondent to report their health status over different recall periods ranging from the current day to the past month. In an ongoing randomised controlled trial in patients with advanced HIV disease, the impact of using a preference-based instrument with a 1-week recall period vs a 1-day recall period (e.g. today) for capturing recently resolved serious adverse events was measured. The results suggest that the instrument with a 1-week recall period gave lower utility values for recently resolved events in comparison with the instrument with a 1-day recall period. A plausible interpretation of these results is that the recall period was adhered to; for example, patients ignored the impact of recently resolved events in their response if the questionnaire asked them only about their health today. While there are limitations to our study, we believe further consideration should be given to the recall period used for preference-based instruments, and future research should examine other patient groups using a single instrument with multiple recall periods. PMID- 18404665 TI - Income, relative income, and self-reported health in Britain 1979-2000. AB - We test the relative income hypothesis that an individual's health depends on the distribution of income in a reference group, as well as on the income of the individual. We use data on 231 208 individuals in Great Britain from 19 rounds of the General Household Survey between 1979 and 2000. Results are insensitive to the measure of self-assessed health used but the sign and significance of the effect of relative income depend on the reference group (national or regional) and the measure of relative income (Gini coefficient, absolute or proportional difference from the reference group mean, Yitzhaki absolute and proportional relative deprivation and affluence). Only one model (relative deprivation measured as income proportional to regional mean income) performs better than the model without relative income and has a positive estimated effect of absolute income on health. In this model the increase in the probability of good health from a ceteris paribus reduction in relative deprivation from the upper quartile to zero is 0.010, whereas an increase in income from the lower to the upper quartile increases the probability by 0.056. While our results provide only very weak support for the relative deprivation hypothesis, the inevitable correlation of measures of individual income and relative deprivation measured by comparing income and incomes in a reference group makes identification of the separate effects of income and relative deprivation problematic. PMID- 18404666 TI - TGF-beta1 and WISP-1/CCN-4 can regulate each other's activity to cooperatively control osteoblast function. AB - Wnt-induced secreted protein-1 (WISP-1), like other members of the CCN family, is expressed in skeletal tissues. Its mechanism of action remains unknown. Expression of WISP-1 was analyzed in human bone marrow stroma cells (hBMSC) by RT PCR. We identified two major transcripts corresponding to those of full-length WISP-1, and of the splice variant WISP-1va which lacks a putative BMP/TGF-beta binding site. To investigate the function of WISP-1 in bone, hBMSC cultures were treated with recombinant human (rh)WISP-1 and analyzed for proliferation and osteogenic differentiation. WISP-1 treatment increased both BrdU incorporation and alkaline phosphatase (AP) activity. Considering the known functional synergy found between the TGF-beta super-family and members of the CCN family, we next tested the effect of WISP-1 on TGF-beta1 activity. We found that rhWISP-1 could reduce rhTGF-beta1 induced BrdU incorporation. Similarly, rhTGF-beta1 inhibited rhWISP-1 induction of AP activity. To explore functional differences between the WISP-1 variants, WISP-1 or WISP-1va were transfected into hBMSC. Both variants could strongly induce BrdU incorporation. However, there were no effects of either variant on AP activity without an additional osteogenic stimulus such as TGF-beta1. Taken together our results suggest a functional relationship between WISP-1 and TGF-beta1. To further define this relationship we analyzed the effect of WISP-1 on TGF-beta signaling. rhWISP-1 significantly reduced TGF-beta1 induced phosphorylation of Smad-2. Our data indicates that full-length WISP-1 and its variant WISP-1va are modulators of proliferation and osteogenic differentiation, and may be novel regulators of TGF-beta1 signaling in osteoblast-like cells. PMID- 18404667 TI - Cytology and receptor architecture of human anterior cingulate cortex. AB - Anterior cingulate cortex (ACC) is involved in emotion, mood, and autonomic regulation. Although a subgenual part of ACC (sACC) may be vulnerable in depression and area 25 is cytologically unique, there are no assessments that contrast this region with pregenual ACC (pACC). Thus, we undertook independent multimodal verifications of architectural differences among subregions and areas. Areas 24a and 24b have pregenual and subgenual components. The latter have a thin layer III. Area 24c has dorsal (pd24c) and ventral (pv24c) parts. Area pd24c has larger neurofilament-expressing neurons in layer Va, and neurons in Vb form aggregates in area pv24c. Area pd24c occupies both banks of the cingulate sulcus, with pv24c on the ventral bank. Layer III of pd24cd has many larger neurofilament expressing neurons and a richer dendritic plexus. Area 32 has pregenual (p32) and subgenual (s32) components. Layer II in s32 is of particular note because it has a neuron-dense IIa and sparse IIb. Area 25 has anterior (25a) and posterior (25p) parts; 25p has the thinnest layer III in the cingulate gyrus. Area 25a contains significantly higher AMPA, kainate, NMDA, GABA(A), GABA(B), and alpha(1) densities than 25p. Area 33 continues around the genu and ventrally to encompass the full caudal extent of area 25. Subgenual ACC has significantly higher GABA(A), GABA(B), benzodiazepine (BZ), alpha(1), and 5-HT(1A) densities than pACC. GABA(B), BZ, and alpha(1) binding confirms the subdivision of area pd24c. In conclusion, ACC comprises two parts that are unique in terms of their cytoarchitecture and neurotransmitter receptor organization. PMID- 18404668 TI - Rhesus glycoprotein and urea transporter genes are expressed in early stages of development of rainbow trout (Oncorhynchus mykiss). AB - The objective of this study was to determine if the genes for the putative ammonia transporters, Rhesus glycoproteins (Rh) and the facilitated urea transporter (UT) were expressed during early development of rainbow trout, Oncorhynchus mykiss Walbaum. We predicted that the Rh isoforms Rhbg, Rhcg1 and Rhcg2 would be expressed shortly after fertilization but UT expression would be delayed based on the ontogenic pattern of nitrogen excretion. Embryos were collected 3, 14 and 21 days postfertilization (dpf), whereas yolk sac larvae were sampled at 31 dpf and juveniles at 60 dpf (complete yolk absorption). mRNA levels were quantified using quantitative polymerase chain reaction and expressed relative to the control gene, elongation factor 1alpha. All four genes (Rhbg, Rhcg1, Rhcg2, UT) were detected before hatching (25-30 dpf). As predicted, the mRNA levels of the Rh genes, especially Rhcg2, were relatively high early in embryonic development (14 and 21 dpf), but UT mRNA levels remained low until after hatching (31 and 60 dpf). These findings are consistent with the pattern of nitrogen excretion in early stages of trout development. We propose that early expression of Rh genes is critical for the elimination of potentially toxic ammonia from the encapsulated embryo, whereas retention of the comparatively benign urea molecule until after hatch is less problematic for developing tissues and organ systems. PMID- 18404669 TI - Regulation of signaling pathways involved in lupeol induced inhibition of proliferation and induction of apoptosis in human prostate cancer cells. AB - Prostate cancer (PCa) is the most frequently diagnosed noncutaneous cancer and the leading cause of cancer related deaths in men in the United States and many other Asian countries. Dietary factors are considered as a strategic agent to control the risk of PCa. Lupeol, a triterpene, present in fruits and medicinal plants, has been shown to possess many pharmacological properties including anticancer effects. Here, effect of lupeol on cell proliferation and cell death was evaluated using human PCa cells, PC-3. In MTT assay, lupeol inhibited the cell proliferation (12-71%) in dose (50-800 microM) and time dependent manner. Flow-cytometric analysis of cell-cycle revealed that an antiproliferative effect of lupeol (400-600 microM) is associated with an increase in G(2)/M-phase arrest (34-58%). RT-PCR analysis showed that lupeol-induced G2/M-phase arrest was mediated through the inhibition of cyclin regulated signaling pathway. Lupeol inhibited the expression of cyclin B, cdc25C, and plk1 but induced the expression of 14-3-3sigma genes. However no changes were observed in the expression of gadd45, p21(waf1/cip1) and cdc2 genes. Results of western blot showed that lupeol regulates the phosphorylation of cdc2 (Tyr15) and cdc25C (Ser198). Further, on increase of lupeol exposure to PC-3 cells an induction of apoptosis was recorded, which was associated with upregulation of bax, caspase-3, -9, and apaf1 genes and down regulation of antiapoptotic bcl-2 gene. The role of caspase-induced apoptosis was confirmed by increase in reactive oxygen species, loss of mitochondrial membrane potential followed by DNA fragmentation. Thus, our study suggests that lupeol possess novel antiproliferative and apoptotic potential against PCa. PMID- 18404670 TI - Comparative analysis of immunohistochemical markers with invasiveness and histologic differentiation in squamous cell carcinoma and basal cell carcinoma of the skin. AB - BACKGROUND: This study evaluates several tumor-related markers to examine the expression pattern of markers according to the invasiveness and histopathologic differentiation of squamous cell carcinoma and basal cell carcinoma. METHODS: Ninety-four cases of squamous cell carcinoma and 108 cases of basal cell carcinoma using tissue array in order to determine correlations between the expression of Ki-67, p53, EGFR, CD44v6, MMP-1 and MMP-3, invasiveness and histologic differentiation. In order to determine invasiveness, we measured the depth of invasion in resected tissues. RESULTS: The depth of invasion showed a correlation with CD44v6 expression of tumor cell in both squamous cell carcinoma and basal cell carcinoma (P = 0.009, P = 0.036, respectively) and with the MMP-1 expression of stromal cell in squamous cell carcinoma (P = 0.010). The differentiation of squamous cell carcinoma was correlated with Ki-67 index. The loss of palisading arrangement in basal cell carcinoma was correlated with the MMP-1 expression of stromal cells (P = 0.045). CONCLUSIONS: CD44v6 and MMP-1, expressed in tumor cells and stromal cells respectively, are significant markers associated with the invasiveness of tumors in squamous cell carcinoma and basal cell carcinoma of the skin and that it will be helpful to evaluate the invasiveness by measuring the expression of these markers. PMID- 18404671 TI - Pooled analysis of the accuracy of five cervical cancer screening tests assessed in eleven studies in Africa and India. AB - Cervical cancer is the main cancer among women in sub-Saharan Africa, India and other parts of the developing world. Evaluation of screening performance of effective, feasible and affordable early detection and management methods is a public health priority. Five screening methods, naked eye visual inspection of the cervix uteri after application of diluted acetic acid (VIA), or Lugol's iodine (VILI) or with a magnifying device (VIAM), the Pap smear and human papillomavirus testing with the high-risk probe of the Hybrid Capture-2 assay (HC2), were evaluated in 11 studies in India and Africa. More than 58,000 women, aged 25-64 years, were tested with 2-5 screening tests and outcome verification was done on all women independent of the screen test results. The outcome was presence or absence of cervical intraepithelial neoplasia (CIN) of different degrees or invasive cervical cancer. Verification was based on colposcopy and histological interpretation of colposcopy-directed biopsies. Negative colposcopy was accepted as a truly negative outcome. VIA showed a sensitivity of 79% (95% CI 73-85%) and 83% (95% CI 77-89%), and a specificity of 85% (95% CI 81-89%) and 84% (95% CI 80-88%) for the outcomes CIN2+ or CIN3+, respectively. VILI was on average 10% more sensitive and equally specific. VIAM showed similar results as VIA. The Pap smear showed lowest sensitivity, even at the lowest cutoff of atypical squamous cells of undetermined significance (57%; 95% CI 38-76%) for CIN2+ but the specificity was rather high (93%; 95% CI 89-97%). The HC2-assay showed a sensitivity for CIN2+ of 62% (95% CI 56-68%) and a specificity of 94% (95% CI 92-95%). Substantial interstudy variation was observed in the accuracy of the visual screening methods. Accuracy of visual methods and cytology increased over time, whereas performance of HC2 was constant. Results of visual tests and colposcopy were highly correlated. This study was the largest ever done that evaluates the cross-sectional accuracy of screening tests for cervical cancer precursors in developing countries. The merit of the study was that all screened subjects were submitted to confirmatory investigations avoiding to verification bias. A major finding was the consistently higher sensitivity but equal specificity of VILI compared with VIA. Nevertheless, some caution is warranted in the interpretation of observed accuracy measures, since a certain degree of gold standard misclassification cannot be excluded. Because of the correlation between visual screening tests and colposcopy and a certain degree of over-diagnosis of apparent CIN2+ by study pathologists, it is possible that both sensitivity and specificity of VIA and VILI were overestimated. Gold standard verification error could also explain the surprisingly low sensitivity of HC2, which contrasts with findings from other studies. PMID- 18404672 TI - Silencing of hSlo potassium channels in human osteosarcoma cells promotes tumorigenesis. AB - Potassium channels, the most diverse superfamily of ion channels, have recently emerged as regulators of carcinogenesis, thus introducing possible new therapeutic strategies in the fight against cancer. In particular, the large conductance Ca(2+)-activated K(+) channels, often referred to as BK channels, are at the crossroads of several tumor-associated processes such as cell proliferation, survival, secretion and migration. Despite the high BK channel expression in osteosarcoma (OS), their function has not yet been investigated in this malignant bone pathology. Here, using stable RNA interference to reduce the expression of hSlo, the human pore-forming alpha-subunit of the BK channel, in human Cal72 OS cells, we show that BK channels play a functional role in carcinogenesis. Our results reveal for the first time that BK channels exhibit antitumoral properties in OS in vivo and affect the tumor microenvironment through the modulation of both chemokine expression and leukocyte infiltration. PMID- 18404673 TI - A comparison of the aldosterone-blocking agents eplerenone and spironolactone. AB - Improved understanding of the adverse pharmacological properties of aldosterone has prompted investigation of the clinical benefits of blocking aldosterone at the receptor level. This article reviews the pharmacology, clinical efficacy, and tolerability of the two available blocking agents, spironolactone and eplerenone. A Medline search identified clinical studies assessing spironolactone and eplerenone. Priority was given to large, well-controlled, clinical trials and comparative studies. Pharmacological differences between spironolactone and eplerenone include lower affinity of eplerenone for progesterone, androgen, and glucocorticoid receptors; more consistently demonstrated nongenomic properties for eplerenone; and the presence of long-acting metabolites for spironolactone. Both agents effectively treat hypertension and heart failure but comparisons are complicated by the deficiency of head-to-head trials and differences between patient populations. There are differences in the tolerability profiles; spironolactone is associated with dose-dependent sexual side effects. Both agents produce dose-dependent increases in potassium concentrations, although the effect with spironolactone appears to be greater when both agents are administered at recommended doses. Choice of a specific agent should be based on individual patient issues, such as the nature of heart failure and patient concerns about adverse events. PMID- 18404674 TI - Hypermethylation of RAS effector related genes and DNA methyltransferase 1 expression in endometrial carcinogenesis. AB - Epigenetic aberration is known to be important in human carcinogenesis. Promoter methylation status of RAS effector related genes, RASSF1A, RASSF2A, hDAB2IP (m2a and m2b regions) and BLU, was evaluated in 76 endometrial carcinomas and their non-neoplastic endometrial tissue by methylation specific PCR. Hypermethylation of at least one of the 5 genes was detected in 73.7% of carcinomas. There were significant correlations between methylation of hDAB2IP and RASSF1A, RASSF2A (p = 0.042, p = 0.012, respectively). Significantly, more frequent RASSF1A hypermethylation was found in Type I endometrioid carcinomas than Type II carcinomas (p = 0.049). Among endometrioid cancers, significant association between RASSF1A hypermethylation and advanced stage, as well as between methylation of hDAB2IP at m2a region with deep myometrial invasion (p < 0.05) was observed. mRNA expression of RASSF1A, RASSF2A and BLU in endometrial cancer cell lines significantly increased after treatment with the demethylating agent 5-Aza 2'-deoxycytidine supporting the repressive effect of hypermethylation on their transcription. Immunohistochemical study of DNMT1 on eight normal endometrium, 16 hyperplastic endometrium without atypia, 40 atypical complex hyperplasia and 79 endometrial carcinomas showed progressive increase in DNMT1 immunoreactivity from normal endometrium to endometrial hyperplasia and endometrioid carcinomas (p = 0.001). Among carcinomas, distinctly higher DNMT1 expression was observed in Type I endometrioid carcinomas (p < 0.001). DNMT1 immunoreactivity correlated with RASSF1A and RASSF2A methylation (p < 0.05). The data suggested that hypermethylation of RAS related genes, particularly RASSF1A, was involved in endometrial carcinogenesis with possible divergent patterns in different histological types. DNMT1 protein overexpression might contribute to such aberrant DNA hypermethylation of specific tumor suppressor genes in endometrial cancers. PMID- 18404675 TI - Pretreatment of docetaxel enhances TRAIL-mediated apoptosis in prostate cancer cells. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising cancer therapeutic agent because of its tumor selectivity. TRAIL is known to induce apoptosis in cancer cells but spare most normal cells. In this study, we examined whether treatment of docetaxel (DTX) can enhance apoptotic cell death by TRAIL against androgen-independent prostate cancer (AIPC). The cell death effect of combinations of TRAIL and docetaxel on prostate cancer cell lines (androgen dependent LNCaP and its derived androgen-independent, metastatic C4-2B) was evaluated by synergisms of apoptosis. Western blot assay and DNA fragmentation assay were used to study the underlying mechanisms of cell death and search for any mechanisms of enhancement of TRAIL induced apoptosis in the presence of docetaxel. In addition, we investigated the in vitro anti-tumor effects of combined docetaxel and TRAIL using MAP kinase inhibitors. Docetaxel itself could not induce apoptotic cell death in 24 h even in high concentration. Apoptotic cell death, however, was drastically enhanced by pretreatment of docetaxel 20 h before TRAIL treatment. Docetaxel enhanced the PARP-1 cleavage and caspases activation by TRAIL especially in androgen-independent, metastatic C4-2B cell line, mainly by phosphorylation of Bcl-2 by JNK activation. It appears that apoptotic cell death was protected by the JNK inhibitor SP600125. The results of our study show that pretreatment of docetaxel is able to enhance the apoptosis produced by TRAIL in prostate cancer cells, especially in hormone-refractory prostate cancer (HRPC). PMID- 18404676 TI - Epigenetics regulate centromere formation and kinetochore function. AB - The eukaryote centromere was initially defined cytologically as the primary constriction on vertebrate chromosomes and functionally as a chromosomal feature with a relatively low recombination frequency. Structurally, the centromere is the foundation for sister chromatid cohesion and kinetochore formation. Together these provide the basis for interaction between chromosomes and the mitotic spindle, allowing the efficient segregation of sister chromatids during cell division. Although centromeric (CEN) DNA is highly variable between species, in all cases the functional centromere forms in a chromatin domain defined by the substitution of histone H3 with the centromere specific H3 variant centromere protein A (CENP-A), also known as CENH3. Kinetochore formation and function are dependent on a variety of regional epigenetic modifications that appear to result in a loop chromatin conformation providing exterior CENH3 domains for kinetochore construction, and interior heterochromatin domains essential for sister chromatid cohesion. In addition pericentric heterochromatin provides a structural element required for spindle assembly checkpoint function. Advances in our understanding of CENH3 biology have resulted in a model where kinetochore location is specified by the epigenetic mark left after dilution of CENH3 to daughter DNA strands during S phase. This results in a self-renewing and self-reinforcing epigenetic state favorable to reliably mark centromere location, as well as to provide the optimal chromatin configuration for kinetochore formation and function. PMID- 18404677 TI - Histone deacetylase 5 represses the transcription of cyclin D3. AB - Histone deacetylases (HDACs) modulate the transcription of a subset of genes by various means. HDAC5 is a class II HDAC whose subcellular location is signal dependent. At present, its known gene targets are few in number. Here we identify a new HDAC5 target: the gene encoding the cell cycle-regulatory protein cyclin D3. When overexpressed in Balb/c-3T3 cells or mouse embryo fibroblasts, HDAC5 substantially reduced the activity of the cyclin D3 promoter and the abundance of endogenous cyclin D3 protein. Conversely, conditions that blocked HDAC5 function increased cyclin D3 expression: treatment of cells with the class I/II HDAC inhibitor trichostatin A (TSA), depletion of HDAC5 from cells by RNA interference, and cytoplasmic sequestration of HDAC5 by co-expression of catalytically active calcium/calmodulin-dependent protein kinase. HDAC5 interacted with the cyclin D3 promoter in vivo, and the HDAC5-responsive element was within 118 base pairs upstream of the transcription start site. Mutation of the Sp1 site and the cyclic AMP response element within this region did not affect the responsiveness of the cyclin D3 promoter to HDAC5 or TSA. PMID- 18404678 TI - Factors influencing collagen biosynthesis. AB - The importance of collagen, the major structural protein of animal kingdom, in maintaining the normal structure and function of the skin is well known. The same property is exploited widely in medical and industrial fields in finding agents, which could influence the synthesis of this protein. In this context in vitro production of collagen is of high significance. A literature survey has been made to analyze the various factors that influence collagen biosynthesis. There are various physical and biological factors that can either induce or inhibit collagen biosynthesis at various levels of gene expression. However reports concentrating on the effects of plants-derived compounds in stimulating collagen synthesis are scanty. Since extracts of many plants are known to be beneficial in the wound healing process, plants-derived compounds will have a definite role in the regulation of collagen synthesis. The present study emphasizes the need for unearthing the role of these plant derived factors on collagen synthesis which will be of immense application in the medical field. PMID- 18404679 TI - Fiber-bound nitrogen in gorilla diets: implications for estimating dietary protein intake of primates. AB - Protein is essential for living organisms, but digestibility of crude protein is poorly understood and difficult to predict. Nitrogen is used to estimate protein content because nitrogen is a component of the amino acids that comprise protein, but a substantial portion of the nitrogen in plants may be bound to fiber in an indigestible form. To estimate the amount of crude protein that is unavailable in the diets of mountain gorillas (Gorilla beringei) in Bwindi Impenetrable National Park, Uganda, foods routinely eaten were analyzed to determine the amount of nitrogen bound to the acid-detergent fiber residue. The amount of fiber-bound nitrogen varied among plant parts: herbaceous leaves 14.5+/-8.9% (reported as a percentage of crude protein on a dry matter (DM) basis), tree leaves (16.1+/-6.7% DM), pith/herbaceous peel (26.2+/-8.9% DM), fruit (34.7+/-17.8% DM), bark (43.8+/ 15.6% DM), and decaying wood (85.2+/-14.6% DM). When crude protein and available protein intake of adult gorillas was estimated over a year, 15.1% of the dietary crude protein was indigestible. These results indicate that the proportion of fiber-bound protein in primate diets should be considered when estimating protein intake, food selection, and food/habitat quality. PMID- 18404680 TI - The evolution of management of acute coronary syndromes (unstable angina and non ST segment elevation myocardial infarction): Part l. AB - As I advanced through medical school at Johns Hopkins, and throughout my subsequent training, I was constantly reminded of the contributions made to medicine by our predecessors. Many of whom were at Johns Hopkins, but many were not. As I continue active involvement in the education of medical students and physicians, it seems to me we have not done a good job of relating the history of how the modern management of patients with cardiovascular disease developed to our students, house staff, and fellows. PMID- 18404681 TI - X-ray burns--painful, protracted, and preventable. AB - Very high doses of x-ray may produce deep burns in the backs of patients having fluoroscopically guided cardiac interventional procedures. While these incidents are uncommon they can be prevented by judicious limitation of fluoroscopy and timely repositioning of the x-ray tube. Better education and improved methods for dose mapping should make these distressing complications a thing of the past. PMID- 18404682 TI - DNA methylation and histone modifications cause silencing of Wnt antagonist gene in human renal cell carcinoma cell lines. AB - Secreted frizzled-related protein 2 (sFRP2) is a negative modulator of the Wingless-type (Wnt) signaling pathway, and shown to be inactivated in renal cell carcinoma (RCC). However, the molecular mechanism of silencing of sFRP2 is not fully understood. Our study was designed to elucidate the silencing mechanism of sFRP2 in RCC. Expression of sFRP2 was examined in 20 pairs of primary cancers by immunohistochemistry. Kidney cell lines (HK-2, Caki-1, Caki-2, A-498 and ACHN) were analyzed for sFRP2 expression using real-time RT-PCR and Western blotting. The methylation status at 46 CpG sites of the 2 CpG islands in the sFRP2 promoter was characterized by bisulfite DNA sequencing. Histone modifications were assessed by chromatin immunoprecipitation (ChIP) assay using antibodies against AcH3, AcH4, H3K4 and H3K9. sFRP2 was frequently repressed in primary cancers and in RCC cells. The majority of sFRP2 negative cells had a methylated promoter. Meanwhile, sFRP2 expression was repressed by a hypomethylated promoter in Caki-1 cells, and these cells had a repressive histone modification at the promoter. In Caki-1 cells, sFRP2 was reactivated by trichostatin A (TSA). Repressive histone modifications were also observed in RCC cells with hypermethylated promoters, but sFRP2 was reactivated only by 5-aza-2'-deoxycytidine (DAC) and not by TSA. However, the activation of the silenced sFRP2 gene could be achieved in all cells using a combination of DAC and TSA. This is the first report indicating that aberrant DNA methylation and histone modifications work together to silence the sFRP2 gene in RCC cells. PMID- 18404683 TI - Estrogen-mediated downregulation of CD24 in breast cancer cells. AB - We have previously reported on the relevance of the prevalence of CD44(+)/CD24( /low) cells in primary breast tumors. To study regulation of CD24, we queried a number of publicly available expression array studies in breast cancer cells and found that CD24 was downregulated upon estrogen treatment. We confirmed this estrogen-mediated repression of CD24 mRNA by quantitative real-time PCR in MCF7, T47D and ZR75-1 cells. Repression was also seen at the protein level as measured by flow cytometry. CD24 was not downregulated in the ER alpha negative MDA-MB-231 cells suggesting that ER alpha was necessary. This was further confirmed by ER alpha silencing in MCF7 cells resulting in increased CD24 levels and by reintroduction of ER alpha into C4-12 cells resulting in decreased CD24 levels. Estrogen treatment did not alter half-life of CD24 mRNA and new protein synthesis was not essential for repression, suggesting a primary transcriptional effect. Histone deacetylase inhibition by Trichostatin A completely abolished the repression, but decrease of the ER alpha corepressors NCoR, LCoR, RIP140, silencing mediator of retinoid and thyroid hormone receptors, SAFB1 and SAFB2 by siRNA or overexpression of SAFB2, NCoR and silencing mediator of retinoid and thyroid hormone receptors had no effect. In silico promoter analyses led to the identification of two estrogen responsive elements in the CD24 promoter, one of which was able to bind ER alpha as shown by electrophoretic mobility shift assay and chromatin immunoprecipitation assay. Together, our results show that CD24 is repressed by estrogen and that this repression is a direct transcriptional effect depending on ER alpha and histone deacetylases. PMID- 18404684 TI - Skin reactions to human papillomavirus (HPV) 16 specific antigens intradermally injected in healthy subjects and patients with cervical neoplasia. AB - We have tested the safety and feasibility of a synthetic long peptide-based HPV16 specific skin test to detect cellular immune responses to HPV16 E2, E6 and E7 in vivo. Women with cervical neoplasia (n = 11) and healthy individuals (n = 19) were intradermally challenged with 8 different pools of HPV16 E2, E6 and E7 peptides. The skin test was safe as the injections were perceived as mildly painful and no adverse events were observed. The majority of skin reactions appeared significantly earlier in HPV16+ patients (<8 days) than in healthy subjects (8-25 days). The development of late skin reactions in healthy subjects was associated with the appearance of circulating HPV16-specific T cells and the infiltration of both HPV16-specific CD4+ Th1/Th2 and CD8+ T cells into the skin. These data show that the intradermal injection of pools of HPV16 synthetic long peptides is safe and results in the migration of HPV16-specific T cells into the skin as well as in an increase in the number of circulating HPV16-specific T cells. The use of this test to measure HPV16-specific immunity is currently tested in a low resource setting for the measurement of spontaneously induced T cell responses as well as in our HPV16 vaccination trials for the detection of vaccine-induced immunity. PMID- 18404685 TI - The sex chromosome that refused to die. AB - Chromosomes that harbor dominant sex determination loci are predicted to erode over time--losing genes, accumulating transposable elements, degenerating into a functional wasteland and ultimately becoming extinct. The Drosophila melanogaster Y chromosome is fairly far along this path to oblivion. The few genes on largely heterochromatic Y chromosome are required for spermatocyte-specific functions, but have no role in other tissues. Surprisingly, a recent paper shows that divergent Y chromosomes can substantially influence gene expression throughout the D. melanogaster genome.1 These results show that variation on Y has an important influence on the deployment of the genome. PMID- 18404686 TI - Musculoskeletal symptoms of the neck and upper extremities among Iowa dairy farmers. AB - BACKGROUND: The prevalence of musculoskeletal symptoms (MSS) among U.S. dairy farmers is relatively unknown. The purpose of this study was to assess the prevalence of neck and upper extremity MSS, and to examine associations between symptoms and dairy operation activities among dairy farmers. METHODS: Questionnaires were mailed to randomly selected dairy farmers in the State of Iowa, USA. Demographic, site specific MSS, and dairy operation activity information was obtained. Odds ratios and 95% confidence intervals were estimated with logistic regression adjusting for potential confounders. RESULTS: Among the participants (N = 341), shoulder MSS were reported most frequently (54%). Neck MSS were significantly associated with manually feeding (OR(adj) = 2.29; 95% CI = 1.09-4.80) and tractors use (OR(adj) = 2.17; 95% CI = 1.05-4.50). Also, wrist/hand MSS were associated with manually cleaning animal stalls (OR(adj) = 1.96; 95% CI = 1.06-3.63). CONCLUSIONS: Neck and upper extremity MSS were common and associated with common dairy farming practices. Future studies need to more accurately assess exposures to physical risk factors for MSS so ergonomic interventions can be developed. PMID- 18404687 TI - Phylogeographic and conservation genetic analysis of the black caiman (Melanosuchus niger). AB - We assessed the spatial distribution of the genetic variability of Melanosuchus niger from 11 localities in South America using 1,027 base pairs of the mitochondrial cytochrome b gene. Screening 132 animals, we found 41 haplotypes, high values of genetic diversity, low values of nucleotide diversity and significant deviations from neutral expectation of allelic frequencies in some localities. Mantel test and nested-clade analysis indicated that isolation-by distance was an important population dynamic for the species as a whole. Wright's fixation indexes analyses showed that hydrogeographically separated populations from French Guiana together with Amapa state population in Brazil are genetically differentiated from all other populations that are found in the Amazon drainage basin. These indexes also disclosed that the population from Ecuador is genetically differentiated in relation to the populations from Brazil, Peru and French Guiana. Within the Amazon Basin little differentiation exists, and genetic and geographic distances are not correlated. Demographic data as well as population genetic data suggest that M. niger is recovering in some protected regions. However, part of this apparent recovery may be owing to the movement of animals into protected regions. PMID- 18404688 TI - Embryonic pattern formation without morphogens. AB - One of the earliest and most-fundamental pattern- formation events in embryonic development is endoderm and mesoderm specification. In sea urchin embryos, this process begins with blimp1 and wnt8 gene expression at the vegetal pole as soon as embryonic transcription begins. Shortly afterwards, wnt8/blimp1 expression spreads to the adjacent ring of mesoderm progenitor cells and is extinguished in the vegetal-most cells. A little later, the ring of wnt8/blimp1 activity moves out of the mesoderm progenitors and into the neighboring endoderm cells. Remarkably, this moving ring of gene expression has now been shown to be controlled entirely by transcriptional cis-regulatory logic.1. PMID- 18404689 TI - Keeping your vascular integrity: What can we learn from fish? AB - The cardiovascular system has the life-providing task of delivering oxygen and any flaw in this system can be life-threatening. This has encouraged extensive studies to elucidate the mechanisms behind cardiovascular development/homeostasis. The zebrafish has emerged as a formidable tool to speed up this quest, as illustrated in a recent issue of Nature Genetics.1 Baculovirus IAP repeat c2 (BIRC2), also termed cellular inhibitor of apoptosis (cIAP)-1, was found to specifically prevent endothelial cells (ECs, lining the inside of vessels) from going into suicide mode ('apoptosis') and so preserve vessel integrity. Here, we summarize the factors determining vascular integrity and elaborate on the suitability of the zebrafish to study this phenomenon. PMID- 18404690 TI - Visualizing new dimensions in Drosophila myoblast fusion. AB - Over several years, genetic studies in the model system, Drosophila melanogastor, have uncovered genes that when mutated, lead to a block in myoblast fusion. Analyses of these gene products have suggested that Arp2/3-mediated regulation of the actin cytoskeleton is crucial to myoblast fusion in the fly. Recent advances in imaging in Drosophila embryos, both in fixed and live preparations, have led to a new appreciation of both the three-dimensional organization of the somatic mesoderm and the cell biology underlying myoblast fusion. PMID- 18404691 TI - On the origins of novelty in development and evolution. AB - The origin of novel traits is what draws many to evolutionary biology, yet our understanding of the mechanisms that underlie the genesis of novelty remains limited. Here I review definitions of novelty including its relationship to homology. I then discuss how ontogenetic perspectives may allow us to move beyond current roadblocks in our understanding of the mechanics of innovation. Specifically, I explore the roles of canalization, plasticity and threshold responses during development in generating a reservoir of cryptic genetic variation free to drift and accumulate in natural populations. Environmental or genetic perturbations that exceed the buffering capacity of development can then release this variation, and, through evolution by genetic accommodation, result in rapid diversification, recurrence of lost phenotypes as well as the origins of novel features. I conclude that, in our quest to understand the nature of innovation, the nature of development deserves to take center stage. PMID- 18404692 TI - Treatment options for muscle-invasive urothelial cancer for patients who were not eligible for cystectomy or neoadjuvant chemotherapy with methotrexate, vinblastine, doxorubicin, and cisplatin: report of Southwest Oncology Group Trial 8733. AB - BACKGROUND: Many patients with invasive urothelial cell cancer are poor candidates for cisplatin-based chemotherapy, and many are high risk for cystectomy. Southwest Oncology Group Trial 8733 was designed to address treatment for such patients. METHODS: Eligible patients had primary or recurrent muscle invasive disease with transitional cell or squamous cell histology, a performance status from 0 to 2, no extrapelvic disease, a life expectancy >3 months, and adequate hematologic function. The treating clinician assigned patients to operable or inoperable groups. All patients received 2 cycles of 5-fluorouracil (5-FU) at a dose of 1000 mg/m(2) per day x 4 starting concurrently with radiation at a dose of 200 centigrays per day x 10 each cycle. After 2 cycles, operable patients with positive biopsies underwent cystectomy, and patients with negative biopsies received a third cycle of chemoradiotherapy. Patients in the inoperable group received 3 cycles without interim biopsy. RESULTS: Eighteen of 24 eligible patients in the operable group were evaluable for response. Five patients had a complete response (CR), 9 patients had stable disease, 1 patient had progressive disease, and 3 patients were not assessable. The median progression-free survival was 10 months (95% confidence interval [95% CI], 4-14 months), and the median overall survival was 18 months (95% CI, 7-28 months). In the inoperable group, 35 of 37 eligible patients were evaluable for response with 17 CRs (49%; 95% CI, 31% 66%). The median progression-free survival was 13 months (95% CI, 10-17 months), and the median overall survival was 20 months (95% CI, 11-53 months). There were no episodes of grade 4 toxicity. CONCLUSIONS: In the current study, the combination of 5-FU and radiation was found to be tolerated well by patients with numerous comorbidities who could not tolerate cisplatin-based therapy or cystectomy. PMID- 18404693 TI - Metabolic syndrome and prostate cancer. PMID- 18404694 TI - Pygopus and the Wnt signaling pathway: a diverse set of connections. AB - Identification of Pygopus in Drosophila as a dedicated component of the Wg (fly homolog of mammalian Wnt) signaling cascade initiated many inquiries into the mechanism of its function. Surprisingly, the nearly exclusive role for Pygopus in Wg signal transduction in flies is not seen in mice, where Pygopus appears to have both Wnt-related and Wnt-independent functions. This review addresses the initial findings of Pygopus as a Wg/Wnt co-activator in light of recent data from both fly and mammalian studies. We compare and contrast the developmental phenotypes of pygopus mutants to those characterized for known Wg/Wnt transducers and explore the data regarding a role for mammalian Pygopus 2 in tumorigenesis. We further analyze the roles of the two conserved domains of Pygopus proteins in transcription, and propose a model for the molecular mechanism of Pygopus function in both Wg/Wnt signaling and Wnt-independent transcriptional regulation. PMID- 18404695 TI - Rate of bilirubin regression after stenting in malignant biliary obstruction for the initiation of chemotherapy: how soon should we repeat endoscopic retrograde cholangiopancreatography? AB - BACKGROUND: This study was conducted to evaluate the rate of regression of bilirubin after stent placement for malignant biliary obstruction. METHODS: Records were reviewed from October 2002 to September 2005 for patients who underwent endoscopic retrograde cholangiopancreatography with stent placement. The time to achieve a bilirubin level or=10 mg/dL (6 weeks) compared with those who had prestent bilirubin levels <10 mg/dL (3 weeks). The following variables were identified as statistically significant: prestent bilirubin level, stricture location, liver metastasis, and INR. The cancer type, recent chemotherapy, stent type and diameter, and sphincterotomy were not statistically significant variables. CONCLUSIONS: The rate of bilirubin normalization after biliary stenting was highly dependent on the prestent bilirubin level. Endoscopic intervention should be considered in patients who fail to achieve adequate normalization of serum bilirubin in 6 weeks if prestent bilirubin level was >or=10 mg/dL and in 3 weeks if their prestent bilirubin level was <10 mg/dL. Independent variables, such as diffuse liver metastases, stricture outside the common bile duct, and elevated INR had predictive value for bilirubin normalization. PMID- 18404696 TI - Thyroid fine-needle aspiration samples inadequate for reverse transcriptase polymerase chain reaction analysis. AB - BACKGROUND: Analysis of different tumor markers by reverse transcriptase polymerase chain reaction (RT-PCR) in fine-needle aspiration samples of thyroid nodules has been studied with the objective of improving the accuracy of the preoperative diagnosis of thyroid lesions. The aim of the current study was to investigate thyroid fine-needle aspiration samples inadequate for RT-PCR analysis and to determine whether there is a correlation between their proportion and the method of sampling used or the greatest dimension of the nodules. METHODS: A total of 350 fine-needle aspiration samples from patients with thyroid nodules were analyzed. After the aspirate was smeared for conventional cytology, the leftover material in the needle was used for RT-PCR analysis in 1 group of 175 patients. In another group of 175 patients, a separate puncture was performed to obtain material for RT-PCR analysis only. Samples were considered adequate for RT PCR analysis if the expression of both glyceraldehyde-3-phosphate dehydrogenase and thyroglobulin was found by RT-PCR. RESULTS: In total, 61 (17.4%) samples inadequate for RT-PCR were detected. All 12 samples that were inadequate for cytologic diagnosis were also found to be inadequate for RT-PCR analysis. The proportion of inadequate samples for RT-PCR was found to be significantly higher in samples taken from leftover material in the needle (21.7%) then in samples from a separate puncture (13.1%) (P = .049). No statistically significant correlation between the adequacy of samples for RT-PCR and the largest dimension of the nodule was found. CONCLUSIONS: The proportion of samples inadequate for RT PCR was found to be higher in samples taken from leftover material in the needle than in samples obtained from a separate puncture. PMID- 18404697 TI - Endoscopic ultrasound and computed tomography in restaging and predicting prognosis after neoadjuvant chemotherapy in patients with locally advanced gastric cancer. AB - BACKGROUND: The objective of the current study was to assess the staging accuracy and prognostic role of preoperative endoscopic ultrasound (EUS) and computed tomography (CT) in patients with locally advanced gastric cancer (LAGC) after neoadjuvant chemotherapy. METHODS: Presurgical LAGC patients underwent EUS and CT before and after 3 cycles of neoadjuvant chemotherapy. Chemotherapy was comprised of docetaxel (at a dose of 36 mg/m(2)) and cisplatin (at a dose of 40 mg/m(2)) on Days 1 and 8 of a 3-week cycle. RESULTS: Forty patients were enrolled in the study. After chemotherapy, the accuracy of EUS and CT was found to be 47% and 57%, respectively for T classification (P = .22) and 39% and 37%, respectively for N classification (P > .99). The 3-year overall survival (OS) rate for patients downstaged with EUS for T and/or N classification was greater than that for nondownstaged patients (69% vs 41%; P = .05). The 2-year recurrence-free survival (RFS) rate was also better for the EUS-downstaged patients than for the nondownstaged patients (77% vs 47%; P = .04). On multivariate analysis, EUS downstaging was found to be correlated with OS (hazards ratio [HR] of 0.12; P = .04), and was correlated with RFS with borderline statistical significance (HR of 0.27; P = .07). The differences in OS and RFS between the patients downstaged with CT and those not downstaged were not found to be statistically significant. CONCLUSIONS: Restaging by EUS and CT after neoadjuvant chemotherapy in patients with LAGC was found to be inaccurate. However, T and/or N downstaging by EUS was found to be correlated with better OS and RFS. Thus, downstaging by EUS may be a useful clinical parameter with which to predict a better outcome for LAGC patients. PMID- 18404698 TI - Comparative in vivo study of six hydroxyapatite-based bone graft substitutes. AB - Improvement of synthetic bone graft substitutes as suitable alternatives to a patient's own bone graft remains a challenge in biomaterials research. Our goal was to answer the question of whether improved osteoinductivity of a material would also translate to better bone-healing orthotopically. Three porous biphasic calcium phosphate (BCP) ceramics (BCPA, BCPB, and BCPC), consisting of hydroxyapatite and beta-tricalcium phosphate, a porous biphasic calcium phosphate ceramic reinforced with a bioresorbable polylactic acid to improve its mechanical properties (BCPC+), a pure hydroxyapatite ceramic (HA), and a carbonated apatite ceramic (CA) were implanted intramuscularly and orthotopically by using a transverse process model in 11 goats for 12 weeks. BCPA and BCPB had similar chemical composition but differed in their microstructure. BCPB was not osteoinductive at all, but BCPA induced ectopic bone formation in 9 of 11 animals. Orthotopically, BCPA performed better than BCPB in both the amount and rate of bone formation. BCPC, similar to BCPA structurally and physicochemically, showed comparable results ectopically and orthotopically. Addition of resorbable polymer to BCPC made the material less osteoinductive (4 of 11 animals) and delayed bone formation orthotopically. Neither HA nor CA were osteoinductive, and their orthotopic performance was inferior to the osteoinductive ceramics. The results of the present study showed that material-derived osteoinduction significantly enhanced bone healing orthotopically, and that this material property appeared more sensitive for predicting orthotopic performance than physicochemical and structural characteristics. PMID- 18404699 TI - Age-adjusted Charlson comorbidity score is associated with treatment decisions and clinical outcomes for patients undergoing radical cystectomy for bladder cancer. AB - BACKGROUND: By using the age-adjusted Charlson comorbidity index (ACCI), the authors characterized the impact of age and comorbidity on disease progression and overall survival after radical cystectomy (RC) for transitional cell carcinoma of the bladder. Also evaluated was whether ACCI was associated with clinicopathologic and treatment characteristics. METHODS: The authors evaluated 1121 patients treated by RC for transitional cell carcinoma of the bladder at a single institution (1990-2004). Logistic regression was used to determine the relation between ACCI and clinical features. They evaluated the association between ACCI and overall and progression-free survival by using multivariate survival-time models with pathologic stage and nodal status as covariates. RESULTS: ACCI scores increased during the study period (P = .009). Extravesical disease was present in 43% of patients with ACCI 5 (P = .051). Despite their higher prevalence of extravesical disease, patients with higher ACCI were less likely to have lymph-node dissection (odds ratio, 0.55 and 0.35, respectively, for ACCI 3-5 and >5 vs 5 vs 1.0 mm) are generally apposed by osseous tissue. Thus in vitro study assessed the mineralization and proliferation response of TF274, MC3T3-E1, murine femoral stromal cells and canine stromal cells to tissue culture plastic (R(a) = 0.001 mm), polished (R(a) = 0.01 mm) and irregularly textured (R(a) = 3.26 mm) titanium surfaces. Amongst all culture systems, proliferation was significantly decreased on textured vs. smooth surfaces. Midway through the culture of the canine marrow cells, the cell layer detached from the tissue culture plastic and polished titanium surfaces. The TF274, MC3T3-E1, murine femoral stromal cell systems formed a mineralized matrix on the tissue culture plastic and polished titanium surfaces which was not observed with the canine stromal cell system. Compared to the tissue culture plastic and polished titanium surfaces, matrix mineralization was significantly reduced on the textured titanium surfaces for the TF274, MC3T3-E1, murine femoral stromal systems, a result which was differed significantly in comparison to the canine stromal system. These results were surprising given the large number of reports concerning the in vivo response to titanium in clinical and pre-clinical studies. Further work is required to determine if the TF274, MC3T3-E1 and murine femoral stromal systems are suitable for the in vitro investigation of the effects of titanium surface texture on osteoblast activity. PMID- 18404721 TI - Complex, multidimensional thumb movements generated by individual extrinsic muscles. AB - The objective of this study was to investigate three-dimensional thumb joint movements produced by individual extrinsic thumb muscles. Ten cadaveric arms were dissected to expose the musculotendinous junctions of the flexor pollicis longus (FPL), abductor pollicis longus (APL), extensor pollicis brevis (EPB), and extensor pollicis longus (EPL). Each muscle/tendon was loaded to 10% of its maximal force capability whereas three-dimensional angular movements of the carpometacarpal (CMC), metacarpophalangeal (MCP), and interphalangeal (IP) joints were obtained simultaneously. We found that each extrinsic muscle produced unique joint angular trajectories in multiple directions. The FPL, APL, EBP, and EPL generated two, two, three, and six movements, respectively. The extrinsic muscles all together generated eight movements among the multiple thumb joints. High interjoint coordination was shown between the MCP joint flexion and IP joint flexion by FPL loading, as well as between the MCP joint extension and IP joint extension by EPL loading. High intrajoint coordination was observed between extension and supination at the CMC joint by the APL, EPL, and EPB. We concluded that each muscle produces movements in multiple joints and/or in multiple anatomical directions. The findings provide a novel insight into the biomechanical roles of the extrinsic muscles of the thumb. PMID- 18404722 TI - Comparison of the healing of open tibial fractures covered with either muscle or fasciocutaneous tissue in a murine model. AB - The objective of this study was to compare the effects of soft tissue coverage by either muscle or fasciocutaneous tissue on the healing of open tibial fractures in a murine model. An open tibial fracture, stripped of periosteum with intramedullary fixation, was created in mice. Experimental groups were devised to allow exclusive comparison of either muscle alone or skin plus fascia in direct contact with healing bone. To exclusively assess the relative efficacy of muscle and fasciocutaneous tissue to promote healing of a fracture stripped of periosteum, a piece of sterile inert material (polytetrafluoroethylene) was positioned anteriorly, excluding skin and fascia (muscle group) or posteriorly, excluding muscle (fasciocutaneous group). Skeletal repair was assessed histologically and quantified by histomorphometry; quantitative peripheral computed tomography (pQCT) and mechanical testing using a four-point bending technique. This standardized, reproducible model allowed characterization of the morphology of open fracture healing. At 28 days postfracture, there was faster healing in the experimental muscle coverage group compared to skin and fascia alone. Furthermore, there was almost 50% more cortical bone content and a threefold stronger union beneath muscle compared to fasciocutaneous tissue (p < 0.05 by one-way ANOVA). Exclusive comparison of muscle and fasciocutaneous tissue in our novel murine model demonstrates that muscle is superior for the coverage of open tibial fractures for both the rate and quality of fracture healing. PMID- 18404723 TI - Changes in gene expression of individual matrix metalloproteinases differ in response to mechanical unloading of tendon fascicles in explant culture. AB - Immobilization of the tendon and ligament has been shown to result in a rapid and significant decrease in material properties. It has been proposed that tissue degradation leading to tendon rupture or pain in humans may also be linked to mechanical unloading following focal tendon injury. Hence, understanding the remodeling mechanism associated with mechanical unloading has relevance for the human conditions of immobilization (e.g., casting), delayed repair of tendon ruptures, and potentially overuse injuries as well. This is the first study to investigate the time course of gene expression changes associated with tissue harvest and mechanical unloading culture in an explant model. Rat tail tendon fascicles were harvested and placed in culture unloaded for up to 48 h and then evaluated using qRT-PCR for changes in two anabolic and four catabolic genes at 12 time points. Our data demonstrates that Type I Collagen, Decorin, Cathepsin K, and MMP2 gene expression are relatively insensitive to unloaded culture conditions. However, changes in both MMP3 and MMP13 gene expression are rapid, dramatic, sustained, and changing during at least the first 48 h of unloaded culture. This data will help to further elucidate the mechanism for the loss of mechanical properties associated with mechanical unloading in tendon. PMID- 18404724 TI - Joint capsule mast cells and neuropeptides are increased within four weeks of injury and remain elevated in chronic stages of posttraumatic contractures. AB - The purpose of this article was to determine mast cell and neuropeptide nerve fiber numbers in joint capsules in posttraumatic contractures, as elevated numbers have been implicated in other fibrotic and contracture conditions. Twelve skeletally mature rabbits had intraarticular cortical windows removed from the medial and lateral femoral condyles and the knee joint immobilized. The contralateral unoperated limb served as a control. Equal numbers of rabbits were sacrificed 4 weeks after surgery or 40 weeks after the first surgery that included 32 weeks of remobilization. Six patients with chronic posttraumatic elbow joint contractures and six age-matched organ donor controls free of elbow contractures were also studied. Joint capsule myofibroblast, mast cell, and neuropeptide containing nerve fiber numbers were assessed with immunohistochemistry. The numbers of myofibroblasts, mast cells, and neuropeptide containing nerve fibers expressed as a percentage of total cells were significantly greater in the contracture capsules when compared to the control capsules at all time points (p < 0.0001). The range of percentages for the three components in the contracture capsules versus the controls were 41-48% versus 9 10%, 44-50% versus 11-13%, and 45-50% versus 10-12% for the acute and chronic stages of the rabbit model and the chronic stages in the human elbows, respectively. These data support the hypothesis that a myofibroblast-mast cell neuropeptide fibrosis axis may underlie some of the pathologic changes in the joint capsule in posttraumatic contractures. Approaches designed to manipulate this axis, such as preventing degranulation of mast cells, warrant further investigation. PMID- 18404725 TI - Enhanced external counterpulsation in the treatment of chronic refractory angina: a long-term follow-up outcome from the International Enhanced External Counterpulsation Patient Registry. AB - BACKGROUND: The management of patients who suffer from medically refractory angina and are unsuitable for conventional revascularization therapy is often unsatisfactory. Enhanced external counterpulsation (EECP) is a noninvasive treatment that is safe and effective immediately after a course of treatment. However, the duration of benefit is less certain. HYPOTHESIS: To evaluate the 3 year outcome of EECP treatment. METHODS: One thousand four hundred and twenty seven patients from 36 centers registered in the International EECP Patient Registry (IEPR)-Phase 1 was prospectively followed for a median of 37 months. Two hundred and twenty patients (15.4%) died, while 1,061 patients (74.4%) completed their follow-up. RESULTS: The mean age was 66+/-11 years and 72% were men. Seventy-six percent had multivessel coronary disease for 11+/-8 years. Eighty eight percent had a prior percutaneous or surgical revascularization and 82% were unsuitable for further coronary intervention. Immediately post-EECP, the proportion of patients with severe angina (Canadian Cardiovascular Angina Classification [CCS] III/IV) were reduced from 89% to 25%, p<0.001. The CCS class was improved by at least 1 class in 78% of the patients and by at least 2 classes in 38%. This was sustained in 74% of the patients during follow-up.Thirty-six percent of the patients had CCS II or less angina, which was better than pre-EECP state without a major adverse cardiovascular event during follow-up. More severe baseline angina and a history of heart failure or diabetes were independent predictors of unfavorable outcome. CONCLUSION: An EECP improves angina and quality of life immediately after a course of treatment. For most of the patients, these beneficial effects are sustained for 3 years. PMID- 18404726 TI - Clinical impact of drug-eluting stents in an unselected population of diabetic patients. AB - BACKGROUND: Drug-eluting stents (DES) have been shown in randomized trials to reduce clinical events in diabetic patients. Our aim was to determine whether these clinical results are applicable in an unselected population of patients with non-insulin-dependent diabetes mellitus (NIDDM) and insulin-dependent diabetes mellitus (IDDM). METHODS: We studied 440 consecutive patients (271 NIDDM and 169 IDDM) who underwent percutaneous coronary intervention, divided into 2 cohorts: Group A (1998-2000): 220 patients with bare metal stents, and Group B (2002-2004): 220 patients with drug-eluting stents. We analyzed major coronary adverse events (death, nonfatal acute myocardial infarction, and target lesion revascularization) over a mean follow-up of 18+/-15 months. RESULTS: Group B had more patients who were insulin-dependent (44.5 versus 32.3% p<0.001) or had hypertension (64.5 versus 54.1%; p=0.02), a lower left ventricular ejection fraction (53.89 versus 56.8%; p=0.04), more complex lesions (B2/C) (82.7 versus 62.3%; p<0.001), more treated lesions (1.40 versus 1.26; p<0.001), more stents implanted (1.69 versus 1.15; p<0.0001), and more patients treated with abciximab (76.8 versus 42.7%; p<0.0001). During the follow-up, Group B had fewer major adverse coronary events (11.7 versus 27.9%; p<0.001) and a reduction in target lesion revascularization (3.9 versus 17.2%; p<0.001), with no differences in death or myocardial infarction. Both groups experienced a significant reduction in events (NIDDM: 8.1 versus 26.7%; p<0.001 and IDDM: 16 versus 31.9%; p=0.016). Multivariate regression analysis showed the use of drug-eluting stents to be in direct relation with event-free survival (odds ratio [OR]: 3.37; 95% confidence interval [CI], 1.44-7.90; p=0.005). CONCLUSION: Despite the worse angiographic characteristics, the use of DES reduced clinical events, particularly target lesion revascularization. PMID- 18404727 TI - Diagnostic approach and treatment strategy in tachycardia-induced cardiomyopathy. AB - BACKGROUND: Due to the absence of differential guidelines for heart failure with tachyarrhythmia, it is difficult to diagnose tachycardia-induced cardiomyopathy (TIC) at the initial visit. Furthermore, clinical outcomes of rate versus rhythm control in TIC are unclear. HYPOTHESIS: Because the etiology of TIC is different from dynamic cardiomyoplasty (DCMP), differential parameters may be present. METHODS: We assessed 21 patients with TIC (15 men; mean age, 50+/-14 years) and 21 control patients with idiopathic DCMP. We assessed clinical courses, echocardiographic parameters, as well as outcomes by treatment. RESULTS: In the TIC group, the related tachyarrhythmias were atrial fibrillation (n=12), atrial flutter (n=5), atrial tachycardia (n=3) and paroxysmal supraventricular tachycardia (n=1). After treatment, all patients became asymptomatic and the ejection fraction (EF) improvement (DeltaEF>or=15%) was observed in all patients (left ventricular ejection fraction [LVEF], 30+/-11%initial versus 58+/-6%last). In the idiopathic DCMP group, no patient showed EF improvement (EF increaseor=3 (P<.0001), a platelet count<100x10(9)/L (P=.004), a monocyte count>or=1x10(9)/L (P=.02), the presence of hypercatabolic symptoms (P=.03), a low hemoglobin level (P=.04), and a high leukocyte count (P=.04). The first 2 parameters were found to maintain their statistical significance during multivariate analysis. Neither leukemia-free nor overall survival was found to be affected by the presence of <3% peripheral blood blasts or JAK2V617F mutation. The evaluation of treatment effect on leukemic transformation unexpectedly revealed a significant and independent association with previous therapy with either erythropoiesis stimulating agents (P=.004) or danazol (P=.007), even when the aforementioned prognostic indicators at the time of diagnosis were added as covariates to the multivariate model. In contrast, leukemia-free survival was not found to be affected by a treatment history with hydroxyurea, thalidomide, or other drugs. CONCLUSIONS: A peripheral blood blast percentage>or=3 and/or a platelet count<100x10(9)/L at the time of diagnosis were found to be strong and independent predictors of leukemic transformation in patients with primary myelofibrosis. The unexpected association between leukemic transformation and a history of treatment with erythropoiesis-stimulating agents or danazol requires validation by prospective studies. PMID- 18404743 TI - Transport processes at alpha-quartz-water interfaces: insights from first principles molecular dynamics simulations. AB - Car-Parrinello molecular dynamics (CP-MD) simulations are performed at high temperature and pressure to investigate chemical interactions and transport processes at the alpha-quartz-water interface. The model system initially consists of a periodically repeated quartz slab with O-terminated and Si terminated (1000) surfaces sandwiching a film of liquid water. At a temperature of 1000 K and a pressure of 0.3 GPa, dissociation of H(2)O molecules into H(+) and OH(-) is observed at the Si-terminated surface. The OH(-) fragments immediately bind chemically to the Si-terminated surface while Grotthus-type proton diffusion through the water film leads to protonation of the O-terminated surface. Eventually, both surfaces are fully hydroxylated and no further chemical reactions are observed. Due to the confinement between the two hydroxylated quartz surfaces, water diffusion is reduced by about one third in comparison to bulk water. Diffusion properties of dissolved SiO(2) present as Si(OH)(4) in the water film are also studied. We do not observe strong interactions between the hydroxylated quartz surfaces and the Si(OH)(4) molecule as would have been indicated by a substantial lowering of the Si(OH)(4) diffusion coefficient along the surface. No spontaneous dissolution of quartz is observed. To study the mechanism of dissolution, constrained CP-MD simulations are done. The associated free energy profile is calculated by thermodynamic integration along the reaction coordinate. Dissolution is a stepwise process in which two Si--O bonds are successively broken. Each bond breaking between a silicon atom at the surface and an oxygen atom belonging to the quartz lattice is accompanied by the formation of a new Si--O bond between the silicon atom and a water molecule. The latter loses a proton in the process which eventually leads to protonation of the oxygen atom in the cleaved quartz Si--O bond. The final solute species is Si(OH)(4). PMID- 18404744 TI - Catalytic properties of nanoscale iron-doped zirconia solid-solution aerogels. AB - Nanoscale iron-doped zirconia solid-solution aerogels are prepared via a simple ethanol thermal route using zirconyl nitrate and iron nitrate as starting materials, followed by a supercritical fluid drying process. Structural characteristics are investigated by means of powder X-ray diffraction (XRD), thermal analyses (TG/DTA), N(2) adsorption measurements and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS). The results show that the resulting iron-doped solid solutions are metastable tetragonal zirconia which exhibit excellent dispersibility and high solubility of iron oxide. Further, when the Fe:(Fe+Zr) ratio x is lower than 0.10, all of the Fe(3+) ions can be incorporated into ZrO(2) by substituting Zr(4+) to form Zr(1-) (x)Fe(x)O(y) solid solutions. Moreover, for the first time, an additional hydroxyl group band that is not present in pure ZrO(2) is observed by DRIFTS for the Zr(Fe)O(2) solid solution. This is direct evidence of Fe(3+) ions incorporated into ZrO(2). These Zr(1-) (x)Fe(x)O(y) solid solutions are excellent catalysts for the solvent-free aerobic oxidation of n-hexadecane using air as the oxidant under ambient conditions. The Zr(0.8)Fe(0.2)O(y) solid-solution catalyst demonstrates the best catalytic properties, with the conversion of n-hexadecane reaching 36.2 % with 48 % selectivity for ketones and 24 % selectivity for alcohols and it can be recycled five times without significant loss of activity. PMID- 18404746 TI - A metallacryptand-based manganese(II)-cobalt(II) ferrimagnet with a three dimensional honeycomb open-framework architecture. PMID- 18404745 TI - Optical analysis of Hg2+ ions by oligonucleotide-gold-nanoparticle hybrids and DNA-based machines. PMID- 18404747 TI - Implications of a carboxylate-bound C-cluster structure of carbon monoxide dehydrogenase. PMID- 18404748 TI - Copper-catalyzed asymmetric propargylic substitution reactions of propargylic acetates with amines. PMID- 18404749 TI - Protection by conformationally restricted mobility: first solid-phase synthesis of triostin A. PMID- 18404750 TI - Catalytic synthesis of beta3-amino acid derivatives from alpha-amino acids. PMID- 18404751 TI - Controlling the growth of charged-nanoparticle chains through interparticle electrostatic repulsion. PMID- 18404752 TI - Oxidative homolysis of a nitrosylchromium complex. AB - Metal(III)-polypyridine complexes [M(NN)(3)](3+) (M = Ru or Fe; NN = bipyridine (bpy), phenanthroline (phen), or 4,7-dimethylphenanthroline (Me(2)-phen)) oxidize the nitrosylpentaaquachromium(III) ion, [Cr(aq)NO](2+), with an overall 4:1 stoichiometry, 4 [Ru(bpy)(3)](3+) + [Cr(aq)NO](2+) + 2 H(2)O --> 4 [Ru(bpy)(3)](2+) + [Cr(aq)](3+) + NO(3)(-) + 4 H(+). The kinetics follow a mixed second-order rate law, -d[[M(NN)(3)](3+)]/dt = nk[[M(NN)(3)](3+)][[Cr(aq)NO](2+)], in which k represents the rate constant for the initial one-electron transfer step, and n = 2-4 depending on reaction conditions and relative rates of the first and subsequent steps. With [Cr(aq)NO](2+) in excess, the values of nk are 283 M(-1) s(-1) ([Ru(bpy)(3)](3+)), 7.4 ([Ru(Me(2)-phen)(3)](3+)), and 5.8 ([Fe(phen)(3)](3+)). In the proposed mechanism, the one-electron oxidation of [Cr(aq)NO](2+) releases NO, which is further oxidized to nitrite, k = 1.04x10(6) M(-1) s(-1), 6.17x10(4), and 1.12x10(4) with the three respective oxidants. Further oxidation yields the observed nitrate. The kinetics of the first step show a strong correlation with thermodynamic driving force. Parallels were drawn with oxidative homolysis of a superoxochromium(III) ion, [Cr(aq)OO](2+), to gain insight into relative oxidizability of coordinated NO and O(2), and to address the question of the "oxidation state" of coordinated NO in [Cr(aq)NO](2+). PMID- 18404753 TI - Optically active mixed phthalocyaninato-porphyrinato rare-earth double-decker complexes: synthesis, spectroscopy, and solvent-dependent molecular conformations. AB - Reaction between the optically active metal-free phthalocyanine with a pi system with noncentrosymmetrical C(2) [corrected] symmetry ((S)- and (R) H(2){Pc(OBNP)(2)}; OBNP=binaphthylphthalocyanine) and half-sandwich complexes [M(III)(acac)(TClPP)] (M=Y, Eu; TClPP=meso-tetrakis(4-chlorophenyl)porphyrinate; acac=acetylacetonate), which were generated in situ from [M(acac)(3)].n H(2)O and H(2)(TClPP) in n-octanol at reflux, provided the first optically active protonated mixed phthalocyaninato-porphyrinato rare-earth double-decker complexes [M(III)H{Pc(OBNP)(2)}(TClPP)] (M=Y, Eu) in good yield. In addition to electronic absorption spectroscopy and magnetic circular dichroism results, circular dichroism shows different spectroscopic features of these mixed-ring rare-earth double-decker compounds in different solvents, such as DMF and CHCl(3), which was well-reproduced on the basis of time-dependent density functional theory calculation results for the yttrium species (S)-[Y(III){Pc(OBNP)(2)}(Por)](-) (Por=porphyrinate, which is obtained by removing the four chlorophenyl groups from the TClPP ligand) in terms of the change in the rotation angle between the two macrocyclic ligands in the double-decker molecules. These results revealed the solvent-dependent nature of the molecular conformation of mixed-ring rare earth double-decker complexes, which suggests a new way of tuning the optical and the electrochemical properties of sandwich-type bis(tetrapyrrole)-metal double decker complexes in solution by changing the solvent. PMID- 18404755 TI - Controlled direct synthesis of C-mono- and C-disubstituted derivatives of [3,3' Co(1,2-C2B9H11)2]- with organosilane groups: theoretical calculations compared with experimental results. AB - Mono- and dilithium salts of [3,3'-Co(1,2-C(2)B(9)H(11))(2)](-), (1(-)), react with different chlorosilanes (Me(2)SiHCl, Me(2)SiCl(2), Me(3)SiCl and MeSiHCl(2)) with an accurate control of the temperature to give a set of novel C(c)-mono- (C(c) = C(cluster)) and C(c)-disubstituted cobaltabis(dicarbollide) derivatives with silyl functions: [1-SiMe(2)H-3,3'-Co(1,2-C(2)B(9)H(10))(1',2' C(2)B(9)H(11))](-) (3(-)); [1,1'-mu-SiMe(2)-3,3'-Co(1,2-C(2)B(9)H(10))(2)](-) (4( )); [1,1'-mu-SiMeH-3,3'-Co(1,2-C(2)B(9)H(10))(2)](-) (5(-)); [1-SiMe(3)-3,3' Co(1,2-C(2)B(9)H(10))(1',2'-C(2)B(9)H(11))](-) (6(-)) and [1,1'-(SiMe(3))(2)-3,3' Co(1,2-C(2)B(9)H(10))(2)](-) (7(-)). In a similar way, the [8,8'-mu-(1'',2'' C(6)H(4))-1,1'-mu-SiMe(2)-3,3'-Co(1,2-C(2)B(9)H(9))(2)](-) (8(-)); [8,8'-mu (1'',2''-C(6)H(4))-1,1'-mu-SiMeH-3,3'-Co(1,2-C(2)B(9)H(9))(2)](-) (9(-)) and [8,8'-mu-(1'',2''-C(6)H(4))-1-SiMe(3)-3,3'-Co(1,2-C(2)B(9)H(9))(1',2' C(2)B(9)H(10))](-) (10(-)) ions have been prepared from [8,8'-mu-(1'',2'' C(6)H(4))-3,3'-Co(1,2-C(2)B(9)H(10))(2)](-) (2(-)). Thus, depending on the chlorosilane, the temperature and the stoichiometry of nBuLi used, it has been possible to control the number of substituents on the C(c) atoms and the nature of the attached silyl function. All compounds were characterised by NMR and UV/Vis spectroscopy and MALDI-TOF mass spectrometry; [NMe(4)]-3, [NMe(4)]-4 and [NMe(4)]-7 were successfully isolated in crystalline forms suitable for X-ray diffraction analyses. The 4(-) and 8(-) ions, which contain one bridging -mu SiMe(2) group between each of the dicarbollide clusters, were unexpectedly obtained from the reaction of the monolithium salts of 1(-) and 2(-), respectively, with Me(2)SiHCl at -78 degrees C in 1,2-dimethoxyethane. This suggests that an intramolecular reaction has taken place, in which the acidic C(c)-H proton reacts with the hydridic Si-H, with subsequent loss of H(2). Some aspects of this reaction have been studied by using DFT calculations and have been compared with experimental results. In addition, DFT theoretical studies at the B3 LYP/6-311G(d,p) level of theory were applied to optimise the geometries of ions 1(-)-10(-) and calculate their relative energies. Results indicate that the racemic mixtures, rac form, are more stable than the meso isomers. A good concordance between theoretical studies and experimental results has been achieved. PMID- 18404754 TI - Tuning the nucleophilicity in cyclopropenylidenes. AB - Cyclopropenylidenes are Huckel aromatic pi systems in which one of the ring atoms is a carbene center. Quantum chemical calculations at the density functional level of theory, supplemented by coupled-cluster calculations, indicate that there is a sizeable energy separation between the lowest-energy singlet and triplet states of these species. Amino groups considerably increase the energy difference between these two states, whereas electron-withdrawing substituents decrease it. The 1,1-dimerization products of cyclopropenylidenes, namely, triafulvalenes, have been investigated. The calculations show that, without steric hindrance and considerable electronic stabilization, cyclopropenylidenes are kinetically unstable and dimerize. Different substituents (alkyl, silyl, terphenyl, amino, and phosphoraneiminato) were probed to tune the frontier orbital energies of cyclopropenylidenes. Accordingly, it is predicted that by a suitable choice of substituents at the olefinic positions, cyclopropenylidenes can be more nucleophilic than their five-membered ring congeners, namely, imidazol-2-ylidenes. PMID- 18404756 TI - An annulation reaction for the synthesis of morpholines, thiomorpholines, and piperazines from beta-heteroatom amino compounds and vinyl sulfonium salts. PMID- 18404757 TI - Total synthesis of macbecin I. PMID- 18404758 TI - An expedient strategy for the synthesis of tryptamines and other heterocycles. PMID- 18404759 TI - Synthesis of diastereomers of complestatin and chloropeptin I: substrate dependent atropstereoselectivity of the intramolecular Suzuki-Miyaura reaction. PMID- 18404760 TI - Analysis of the tetronomycin gene cluster: insights into the biosynthesis of a polyether tetronate antibiotic. AB - The biosynthetic gene cluster for tetronomycin (TMN), a polyether ionophoric antibiotic that contains four different types of ring, including the distinctive tetronic acid moiety, has been cloned from Streptomyces sp. NRRL11266. The sequenced tmn locus (113 234 bp) contains six modular polyketide synthase (PKS) genes and a further 27 open-reading frames. Based on sequence comparison to related biosynthetic gene clusters, the majority of these can be assigned a plausible role in TMN biosynthesis. The identity of the cluster, and the requirement for a number of individual genes, especially those hypothesised to contribute a glycerate unit to the formation of the tetronate ring, were confirmed by specific gene disruption. However, two large genes that are predicted to encode together a multifunctional PKS of a highly unusual type seem not to be involved in this pathway since deletion of one of them did not alter tetronomycin production. Unlike previously characterised polyether PKS systems, oxidative cyclisation appears to take place on the modular PKS rather than after transfer to a separate carrier protein, while tetronate ring formation and concomitant chain release share common mechanistic features with spirotetronate biosynthesis. PMID- 18404761 TI - Light-regulated RNA-small molecule interactions. PMID- 18404762 TI - Photocontrol of single-chain DNA conformation in cell-mimicking microcompartments. PMID- 18404763 TI - Understanding the supramolecular self-assembly of the fullerene derivative PCBM on gold surfaces. AB - We report a DFT study on the self-assembly of the fullerene derivative PCBM on the Au(111) surface. Recent STM experiments (Angew. Chem. 2007, 119, 8020 8023([1])) show a coverage-dependent transition of the adsorption and self assembly of PCBM on this surface. To understand the origin of this observation, we compute the geometries and relative energies of ten PCBM dimers and four tetramers. The calculations show that the self-assembly of PCBM at high coverage is mainly controlled by hydrogen bonding between the PCBM tails. Due to the large size of the fullerene cage, the hydrogen bonds are formed far away from the surface; hence they are very similar to those found in the gas phase. This picture successfully explains the observed site-insensitive adsorption at high coverage and the 2D arrangement of PCBM on the surface. PMID- 18404764 TI - Kinetic gate-opening process in a flexible porous coordination polymer. PMID- 18404765 TI - Azadipeptide nitriles: highly potent and proteolytically stable inhibitors of papain-like cysteine proteases. PMID- 18404766 TI - Cholesterol seco-sterol-induced aggregation of methylated amyloid-beta peptides- insights into aldehyde-initiated fibrillization of amyloid-beta. PMID- 18404767 TI - Color change of proteorhodopsin by a single amino acid replacement at a distant cytoplasmic loop. PMID- 18404768 TI - Mechanistic kinetic model for symmetric carboligations using benzaldehyde lyase. AB - For reactions using thiamine diphosphate (ThDP)-dependent enzymes many empirically-derived kinetic models exist. However, there is a lack of mechanistic kinetic models. This is especially true for the synthesis of symmetric 2-hydroxy ketones from two identical aldehydes, with one substrate acting as the donor and the other as the acceptor. In this contribution, a systematic approach for deriving such a kinetic model for thiamine diphosphate (ThDP)-dependent enzymes is presented. The derived mechanistic kinetic model takes this donor-acceptor principle into account by containing two K(m)-values even for identical substrate molecules. As example the stereoselective carbon-carbon coupling of two 3,5 dimethoxy-benzaldehyde molecules to (R)-3,3',5,5'-tetramethoxy-benzoin using benzaldehyde lyase (EC 4.1.2.38) from Pseudomonas fluorescens is studied. The model is derived using a model-based experimental analysis method which includes parameter estimation, model analysis, optimal experimental design, in silico experiments, sensitivity analysis and model revision. It is shown that this approach leads to a robust kinetic model with accurate parameter estimates and an excellent prediction capability. PMID- 18404769 TI - Quantification of power consumption and oxygen transfer characteristics of a stirred miniature bioreactor for predictive fermentation scale-up. AB - Miniature parallel bioreactors are becoming increasingly important as tools to facilitate rapid bioprocess design. Once the most promising strain and culture conditions have been identified a suitable scale-up basis needs to be established in order that the cell growth rates and product yields achieved in small scale optimization studies are maintained at larger scales. Recently we have reported on the design of a miniature stirred bioreactor system capable of parallel operation [Gill et al. (2008); Biochem Eng J 39:164-176]. In order to enable the predictive scale-up of miniature bioreactor results the current study describes a more detailed investigation of the bioreactor mixing and oxygen mass transfer characteristics and the creation of predictive engineering correlations useful for scale-up studies. A Power number of 3.5 for the miniature turbine impeller was first established based on experimental ungassed power consumption measurements. The variation of the measured gassed to ungassed power ratio, P(g)/P(ug), was then shown to be adequately predicted by existing correlations proposed by Cui et al. [Cui et al. (1996); Chem Eng Sci 51:2631-2636] and Mockel et al. [Mockel et al. (1990); Acta Biotechnol 10:215-224]. A correlation relating the measured oxygen mass transfer coefficient, k(L)a, to the gassed power per unit volume and superficial gas velocity was also established for the miniature bioreactor. Based on these correlations a series of scale-up studies at matched k(L)a (0.06-0.11 s(-1)) and P(g)/V (657-2,960 W m(-3)) were performed for the batch growth of Escherichia coli TOP10 pQR239 using glycerol as a carbon source. Constant k(L)a was shown to be the most reliable basis for predictive scale-up of miniature bioreactor results to conventional laboratory scale. This gave good agreement in both cell growth and oxygen utilization kinetics over the range of k(L)a values investigated. The work described here thus gives further insight into the performance of the miniature bioreactor design and will aid its use as a tool for rapid fermentation process development. PMID- 18404770 TI - Tuning the expression level of recombinant proteins by modulating mRNA stability in a cell-free protein synthesis system. AB - In this work, it was discovered that the stability of mRNA in a cell-free extract could be controlled by using engineered T7 terminator sequences. Specifically, it was found that mRNA stability gradually decreased as the length of the stem structure of the T7 terminator was reduced sequentially. As a result of the controlled abundance of mRNA species, it was possible to manipulate the relative expression level of target proteins by employing the T7 terminator of adjusted stem lengths. PMID- 18404771 TI - Competitive activation of C-H and C-X bonds in reactions of Pt+ with CH3X (X=F,Cl): experiment and theory. AB - The reactions of Pt(+) with CH(3)X (X=F, Cl) are studied experimentally by employing an inductively coupled plasma/selected-ion flow tube tandem mass spectrometer and theoretically by density functional theory. Dehydrogenation and HX elimination are found to be the primary reaction channels in the remarkably different ratios of 95:5 and 60:40 in the fast reactions of Pt(+) with CH(3)F and CH(3)Cl, respectively. The observed kinetics are consistent with quantum chemistry calculations, which indicate that both channels in the reaction with CH(3)F are exothermic with ground-state Pt(+)((2)D), but that HF elimination is prohibited kinetically because of a transition state that lies above the reactant entrance. The observed HF-elimination channel is attributed to a slow reaction of CH(3)F with excited-state Pt(+)((4)F) for which calculations predict a small barrier. The calculations also show that both the HCl-elimination and dehydrogenation channels observed with CH(3)Cl are thermodynamically and kinetically allowed, although the state-specific product distributions could not be ascertained experimentally. Further CH(3)F addition is observed with the primary products to produce PtCH(2) (+)(CH(3)F)(1,2) and PtCHF(+)(CH(3)F)(1,2). With CH(3)Cl, sequential HCl elimination is observed with PtCH(2) (+) to form PtC(n)H(2) (n) (+) with n=2, 3, which then add CH(3)Cl sequentially to form PtC(2)H(4) (+)(CH(3)Cl)(1-3) and PtC(3)H(6) (+)(CH(3)Cl)(1,2). Also, sequential addition is observed for PtCHCl(+) to form PtCHCl(+)(CH(3)Cl)(1,2). PMID- 18404774 TI - Synthesis and antiviral evaluation of novel 5-(N-Aryl-aminomethyl-1,3,4-oxadiazol 2-yl)hydrazines and their sugars, 1,2,4-triazoles, tetrazoles and pyrazolyl derivatives. AB - A number of new N-arylaminomethyl-1,3,4-oxadiazole derivatives 2, 3a,b, and 9 12a,b were prepared. Sugar (5-N-arylaminomethyl-1,3,4-oxadiazol-2-yl) hydrazones 4-6a,b were synthesized by the reaction of the hydrazino derivatives 3a,b with the corresponding monosaccharides. The novel acyclo-C-nucleosides 7, 8a,b were prepared by heterocyclization of the sugar hydrazones 4, 5a,b with acetic anhydride. A number of the synthesized compounds were tested for their antiviral activity against herpes simplex virus type-1 (HSV-1) and hepatitis-A virus (HAV, MBBcell culture-adapted strain). The results revealed that the sugar hydrazones 6a,b showed higher antiviral activity compared to the other hydrazones and their acetylated derivatives. PMID- 18404775 TI - Stable non-IPR C60 and C70 fullerenes containing a uniform distribution of pyrenes and adjacent pentagons. AB - The most abundant fullerenes, C(60) and C(70), and all the pure carbon fullerenes larger than C(70), follow the isolated-pentagon rule (IPR). Non-IPR fullerenes containing adjacent pentagons (APs) have been stabilized experimentally in cases where, according to Euler's theorem, it is topologically impossible to isolate all the pentagons from each other. Surprisingly, recent experiments have shown that a few endohedral fullerenes, for which IPR structures are possible, are stabilized in non-IPR cages. We show that, apart from strain, the physical property that governs the relative stabilities of fullerenes is the charge distribution in the cage. This charge distribution is controlled by the number and location of APs and pyrene motifs. We show that, when these motifs are uniformly distributed in the cage and well-separated from one other, stabilization of non-IPR endohedral and exohedral derivatives, as well as pure carbon fullerene anions and cations, is the rule, rather than the exception. This suggests that non-IPR derivatives might be even more common than IPR ones. PMID- 18404776 TI - Synthesis and biological activity of functionalized indole-2-carboxylates, triazino- and pyridazino-indoles. AB - Condensation of aryl hydrazines with ethyl pyruvate gave the respective hydrazones 4-6; Fischer indolization led to substituted-1H-indole-2-carboxylic acid ethyl esters 7-9. The Mannich reaction of these compounds with formaldehyde and morpholine yielded ethyl 3-(morpholinomethyl)-substituted-1H-indole-2 carboxylates 10-12. The 5,7-dichloro-1H-indole-2-carbohydrazide 13 was cyclized with methyl orthoformate in DMF to give 6,8-dichloro[1,2,4]triazino[4,5-a]indol 1(2H)-one 14. Vilsmeier-Haack formylation of 7-9 gave ethyl 3-formyl-substituted 1H-indole-2-carboxylates 15-17 whose 2,2'-((5-chloro-2-(ethoxycarbonyl)-1H-indol 3-yl)methylene)bis-(sulfanediyl) diacetic acid 18 was prepared. The reaction of 15 and 16 with substituted anilines by conventional and microwave methods gave ethyl 3-(N-aryliminomethyl)-5-halo-1H-indole-2-carboxylates 19-29. In a cyclocondensation reaction of 19-25 with thiolactic acid or thioglycolic acid substituted indolylthiazolidinones 30-33 were prepared. Reaction of hydrazine hydrate with 15-17 did not give the respective hydrazones but directly led to the cyclized products substituted-3H-pyridazino[4,5-b]indol-4(5H)-ones 34-36, while a reaction with 2,4-dichlorophenylhydrazine yielded the uncyclized hydrazones. The chlorination of 35 and 36 with POCl3 gave pyridazino[4,5-b]indoles 39 and 40, respectively; reaction of the latter compounds with morpholine gave 4 (substituted-5H-pyridazino[4,5-b]indol-4-yl)morpholine 41 and 42. Mannich reaction of 34 with formaldehyde and N-ethylpiperazine gave 8-chloro-3-((4 ethylpiperazin-1-yl)methyl)-3H-pyridazino[4,5-b]indol-4(5H)-one 43. The microwave assistance of selected reactions has a profound effect on the reaction speed. The structures of the new compounds were confirmed by both analytical and spectral data. Some compounds were subjected to investigations concerning their antimicrobial, tranquilizing, and anticonvulsant activities. PMID- 18404778 TI - Synthesis and anticancer activity of new 1-substituted-6H-pyrido[4,3-b]carbazole derivatives. AB - This study examines the synthesis and cytostatic activity of new 5,6-dimethyl-1 substituted-6H-pyrido[4,3-b]carbazole derivatives. Their structures were confirmed by (1)H-NMR and elemental analysis. Seven of the new compounds were tested by the SRB method in vitro against human lung cancer (A549) and human kidney cancer (A498) cell lines. Biological tests indicated remarkable cytostatic effects of four compounds tested in comparison with ellipticine and cisplatin as reference drugs. One particular compound 3c was about four times more active on A498 than ellipticine with similar activity on the A549 cell line, and outperformed cisplatin activity on both tumor cell lines. PMID- 18404777 TI - Synthesis, cytotoxicity and protein kinase C inhibition of arylpyrrolylmaleimides. AB - A series of novel arylpyrrolylmaleimides was synthesized and evaluated for their in-vitro cytotoxicity against various human cancer cell lines and their protein kinase C inhibitory activity. Some of the compounds showed high or moderate cytotoxic activity against the tested cell lines. Compound 6b is the most promising compound against the tested cancer cell lines; 6d and 6e showed moderate protein-kinase C inhibition. Structure-activity relationships are discussed based on the experimental data obtained. PMID- 18404779 TI - Investigations concerning the correlation of COX-1 inhibitory and hydroxyl radical scavenging activity. AB - The aim was to study the COX-1 inhibiting efficacy in context with hydroxyl radical scavenging properties of compounds bearing a carboxylic acid and ester function, respectively. In general, the acids are more potent radical scavengers than the corresponding esters but there is no clear correlation with their COX-1 inhibiting potencies. A feasible scavenging mechanism of carboxylic acids is discussed. PMID- 18404780 TI - Erythropoietin in cancer: the new face of an old friend. AB - During the last decade, anemia, a very common situation in patients with malignant diseases, either associated with chemotherapy or not, is being treated with recombinant erythropoietin (rEPO). Recent experimental findings have elucidated the role of EPO as a strongly anti-apoptotic agent in multiple non erythroid and neoplastic tissues. The discovery of probably functional EPO receptors (EPOR) on malignant cells, hinting that EPO may act as a tumor growth factor, raised embarrassing thoughts regarding the routine administration of erythropoiesis-stimulating agents (ESAs). In addition, the results of a few clinical trials showing a negative impact on overall survival of rEPO-treated cancer patients, although strongly criticized for several methodological pitfalls, led the FDA to force a "black label" warning concerning the use of rEPO and to recommend that physicians should use the lowest possible dose of ESAs in chemotherapy-treated cancer patients. This recommendation comes in accord with the recent guidelines of European Organisation for Research and Treatment of Cancer (EORTC) which are reviewed in this paper, along with the structure of EPO and EPOR, the role of EPO on normal and malignant cells and the clinical applications of EPO. PMID- 18404781 TI - Cell signaling in cancer. AB - The pathways that control cell differentiation and growth are almost always compromised in cancer. Although in the last years there have been major advances in understanding these changes and how they contribute to tumor initiation and growth, the task is far from complete. In this review we discuss some of the factors that are found in major key nodes of the signaling pathways. Included among them are the nuclear factor kappaB (NF-kappaB) that has a quite central role in inflammation, and the mammalian target of rapamycin (mTOR) kinase. Also an eminent role is played by the EGF (epidermal growth factor) and the Snail/Slug family of repressors. Since the expansion of tumor cells depends heavily on nutrient and oxygen supply, it requires the growth of new blood vasculature which is directed by the hypoxia inducible factor (HIF). PMID- 18404782 TI - Risk factors and adjuvant chemotherapy in the treatment of endometrial cancer. AB - Endometrial carcinoma is the most common and potentially curable gynecologic malignant neoplasm. The staging of endometrial cancer, according to the International Federation of Gynecology and Obstetrics (FIGO), is surgical. Recent studies suggest a therapeutic benefit associated with extensive retroperitoneal lymph node evaluation to determine the disease extent and thereby more effectively direct potentially life-saving adjuvant therapy. Due to the increasing number of endometrial cancer patients who undergo surgical staging, some independent prognostic factors have been identified in early stages (stage I II), including lymph-vascular space involvement, histologic grade 3, aggressive histologic subtypes (uterine papillary serous carcinoma, clear cell carcinoma), depth of myometrial invasion, cervical invasion and the age of patients. Adjuvant radiation therapy, known to offer survival benefit in advanced-stage disease, may also offer survival benefit in intermediate-risk surgical stage I, but this is followed by a significant risk of serious complications. Based on randomized clinical trials, this review identified that only a limited body of evidence is available which can help clinicians make decisions about adjuvant chemotherapy of patients with high-risk stage I and II, as well as stage IIIA endometrial cancer. Further investigations are required to define the subgroup of patients who benefit from postoperative adjuvant chemotherapy. In addition, the optimal regimen remains to be defined as all of them (doxorubicin/cisplatin--AP, cyclophosphamide/ doxorubicin/cisplatin--CAP, paclitaxel/carboplatin--TC, paclitaxel/doxorubicin/cisplatin--TAP) cause significant toxicity. Thereby, combination of carboplatin plus paclitaxel represents an efficacious, low toxicity regimen for managing intermediate-risk surgical stage I, as well as advanced or recurrent endometrial cancer. PMID- 18404783 TI - Chemotherapy and cerebrovascular disease. AB - Cerebrovascular disease is common in cancer patients. Some tumors are at high risk for cerebrovascular complications. The development of cerebrovascular disease may be provoked by cancer treatment. No well-planned prospective studies about other causes of thrombosis are available, although various case reports about thrombosis related to chemotherapy have been published. L-asparaginase, cisplatin, 5-fluorouracil (5-FU) and methotrexate are anticancer agents which are reported to relate to stroke. The mechanisms by which antineoplastic agents may lead to stroke include endothelium toxicity and abnormalities of coagulation factors. Also, brain hemorrhages that could result from chemotherapy effects on the hemostatic system were reported. Besides, it is difficult to determine whether stroke is caused by chemotherapy or cancer itself. Clinicians deal not only with problems originating from cancer itself, but also with the complications resulting from its treatment. Treatment-induced cerebrovascular disorders affect quality of life and survival in cancer patients. For this reason, cancer treatment should be planned by taking into consideration the possibility of cerebrovascular complications. PMID- 18404784 TI - Capecitabine as salvage treatment in relapsed nasopharyngeal carcinoma: a phase II study. AB - PURPOSE: To evaluate the efficacy of capecitabine (Xeloda) as rescue treatment (2nd, 3rd and 4th line) in patients with relapsed nasopharyngeal carcinoma (NPC) in a phase II study. PATIENTS AND METHODS: Between 5/2002-11/2005, 23 relapsed NPC patients (17 locoregional relapse, 3 metastatic, 3 locoregional + metastatic) received capecitabine 2500 mg/m(2)/d, days 1-14 every 3 weeks, until progression or for a maximum of 6 cycles. RESULTS: PATIENT CHARACTERISTICS: 23 patients (14 men, 9 women) with median age 46 years (range 15-59); ECOG performance status 1 n=21, 2 n=2; histology: undifferentiated carcinoma (WHO type III) n=21, non keratinizing epidermoid carcinoma (WHO type II), n=2. Capecitabine was given as 2nd--(13 patients), 3rd--(7 patients), and 4th--(3 patients) line chemotherapy. Previous chemotherapy regimes were epirubicin + cisplatin, paclitaxel + carboplatin, paclitaxel + 5-fluorouracil and leucovorin (5-FU/LV) or methotrexate. 104 cycles were given (median 5, range 2-6). Two (9%) patients achieved complete response (CR); 9 (39%) partial response (PR); 9 (39%) stable disease (SD) and 3 (13%) progressed (PD). Toxicity was mild without toxic deaths or grade 4 toxicities. The most frequent toxicities (grades 1-3) were anemia (38%), hand-foot syndrome (23%), leukopenia (13%) and diarrhea (7%). Median follow-up was 10 months (range 2-44). Median overall survival was not reached at 18 months and actuarial one-year survival was 62% (95% confidence interval/CI: 41 80). Median progression-free survival was 14 months. CONCLUSION: Capecitabine is active in relapsed NPC patients, achieving 48% objective responses, with mild toxicity. It is an attractive therapy to be administered in an outpatient setting. PMID- 18404785 TI - Toxicity and feasibility analysis for cisplatin-based concomitant chemoradiotherapy in locally advanced nasopharyngeal carcinoma. AB - PURPOSE: To assess the side effects of cisplatin-based concurrent chemoradiotherapy (CRT) for locally advanced nasopharyngeal carcinoma (NPC). PATIENTS AND METHODS: From 2001 through 2007, 34 (27 males; 7 females) patients received a median of 70 Gy curative radiotherapy (RT) with conventional fractionation. Twenty-one (62%) patients received induction chemotherapy (CT): 8 of them received 2 courses of cisplatin (75 mg/m(2), day 1) and 5-fluorouracil (5 FU) (750 mg/m(2), days 1-5) every 3 weeks and 13 patients received 3 courses of cisplatin (75 mg/m(2), day 1) and docetaxel (75 mg/m(2), day 1) every 3 weeks. Concomitant cisplatin was administered either 40 mg/m(2) weekly (n=8) or 75-80 mg/m(2) every 3 weeks (n=26) during RT. Median Karnofsky performance status (KPS) prior to RT was 80 (range 70-90). Patient, disease and treatment-related factors were analysed in relation to termination of concurrent CT. RESULTS: Concurrent CT was administered to 13 (38.2%) patients without cisplatin termination, whereas 10 (29.4%) patients received 2 cycles of the 3-weekly schedule. Grade 3 oral mucositis (47.1%), grade 2-3 weight loss (44.2%) and grade 2 fatigue (44.1%) were the most frequently dose-limiting side effects during concurrent therapy. The rate of receiving cisplatin cycles as planned was 85% for patients with KPS >80, whilst it was 15% only for patients with KPS < or = 80 (p=0.006). None of the patients suffering of grade 2 fatigue could complete all cycles compared to 68% of patients with < grade 2 fatigue who completed all cycles (p <0.001). The severity of mucositis was significantly related to initial haemoglobin level (p=0.02) and weight loss during RT (p=0.04). Median follow-up was 20 months (range 5-65). Three-year locoregional relapse free (LRRFS), disease free (DFS) and overall survival (OS) rates were 79.3%, 68.8% and 79.2%, respectively. CONCLUSION: Concurrent administration of CT during RT reveals better outcome but requires careful consideration for toxicity. Initial performance status prior to CRT might be a predictor for unplanned CT stopping due to side effects. PMID- 18404786 TI - Clinical staging and histopathological findings after radical hysterectomy in FIGO stage IIB cervical cancer. AB - PURPOSE: Advanced cervical cancer still represents a major health care challenge in the developing world. According to standard protocols the treatment of choice for stage IIB cervical cancer is cisplatin-based chemoradiotherapy. However, in some European countries, and especially in Japan, patients with stage IIB cervical cancer are generally treated with radical hysterectomy as initial treatment. The aim of this study was to compare clinical stage with pathological findings, and also to correlate any relationship between parametrial infiltration and nodal status. PATIENTS AND METHODS: From 1997 to 2006, 26 patients with FIGO stage IIB cervical cancer were radically operated (Piver class III operation). Preoperative clinical findings were compared with the pathological findings of the surgical material. The correlation between infiltration of the parametria and lymph node status was also examined. Fisher's exact test was used to examine statistical significance. RESULTS: The patients' median age was 48.3 years (range 36-61). The median number of removed lymph nodes was 16 (range 8-40). The histopathological types of cervical tumors were: squamous cell carcinoma 80%, adenosquamous carcinoma 15% and adenocarcinoma 5%. In 50% of the patients the parametria were infiltrated, suggesting that 50% of the patients were clinically overstaged. Positive lymph nodes were found in 69% of patients with positive parametria and 15% in patients with negative parametria (p <0.05). Patients with positive lymph nodes received adjuvant chemoradiotherapy. CONCLUSION: Adequate preoperative staging such as clinical examination under anesthesia or nuclear magnetic resonance could help to exclude parametrial involvement in equivocal cases. Parametrial invasion presents an important risk factor for lymph nodes metastases. PMID- 18404788 TI - Factor V 1691 G-A (Leiden) polymorphism and cancer-related venous thromboembolism: a meta-analysis of published studies. AB - PURPOSE: Although one of the most common genetic polymorphisms that predispose to venous thromboembolism (VTE) is factor V 1691 G-A (Factor V Leiden; FVL), the effect of this polymorphism on the development of VTE in cancer patients is unclear. We have therefore performed a meta-analysis to estimate the risk of VTE associated with FVL among cancer patients. MATERIALS AND METHODS: Relevant studies published before May 2006 were retrieved from Pubmed/Medline. We selected studies comparing the prevalence of FVL in cancer patients with VTE with cancer patients without VTE. Both fixed and random effect models were used. P-values and odds ratios (ORs) with 95% confidence intervals (95% CI) were calculated by Fisher's exact test. RESULTS: Pooled results from 9 studies, comprising 397 cancer patients with VTE and 678 cancer patients without VTE revealed that the prevalence of FVL was higher among cancer patients with VTE (7.3%) than those without VTE (4.6%) (chi(2) = 34.633 > chi(2) (18; 0.05) =28.869) (p=0.013). It was also found that mean effect size of FVL was 0.22 (95% CI 0.051-0.4892). Using the homogeneity test, there was evidence of statistical heterogeneity (Q(hom)= 46.334> chi(2) (8; 0.05) =15.507) (p=0.0000). CONCLUSION: Although the published studies were small and the meta-analysis demonstrated an association of FVL with cancer-related thrombosis, testing for FVL can be routinely used as part of clinical thrombophilia assessment in cancer patients from the regions with high incidence of FVL. PMID- 18404787 TI - Impact of body mass index on cancer development. AB - PURPOSE: To determine the impact of body mass index (BMI) on cancer in a hospital based Turkish population. PATIENTS AND METHODS: The study group consisted of 2015 (1172 females: 423 pre- and 749 postmenopausal; and 843 males) patients with histologically proven cancer who applied to Marmara University Medical School, Medical Oncology Clinic. The control group included 305 healthy caregivers (192 females: 110 pre- and 82 postmenopausal; and 113 males). RESULTS: Mean BMI of the patients with breast, ovarian and cervical carcinoma was significantly higher than that of the healthy female controls (p<0.001, 0.003, <0.001, respectively). Postmenopausal breast cancer patients had significantly higher BMI than postmenopausal female controls (odds ratio [OR] 1.3; 95% confidence interval [CI], 1.06-1.6; p=0.012), while this was not seen in premenopausal patients. When compared with controls obese postmenopausal female patients had 3.26-fold (95% CI 1.54-6.90) increased risk for breast cancer (p=0.002). Mean BMI of lung, stomach, esophagus, pancreas and head and neck carcinoma patients was significantly lower than that of the healthy controls. Female patients with lung and colorectal carcinoma had higher BMI than female controls. CONCLUSION: Elevated BMI might be a risk factor for breast cancer in postmenopausal women. Case-control studies may not show the actual association between BMI and cancers that present with pre diagnosis weight loss and advanced stage. PMID- 18404789 TI - Pressure dressing in breast surgery: is this the solution for seroma formation? AB - PURPOSE: Pressure dressing (PD) after modified radical mastectomy (MRM) for breast cancer is investigated here as an easy-to-apply method to reduce seroma formation and subsequent need for clinical care. PATIENTS AND METHODS: Two hundred mastectomized patients who were treated with PD on the skin flaps and the axilla immediately postoperatively (group A) were compared with a similar non-PD group (B). Surgical technique and perioperative care were the same. Drains were removed when drain output was reduced to 30 ml per day, or on postoperative day 8 regardless of output. RESULTS: Mean time with drains kept in situ was 4.9 and 5.5 days in group A and B, respectively. Five (2.5%) patients in group A and 16 (8%) in group B developed seromas after the removal of the drains. In total, 9 seroma needle aspirations were performed in group A and 26 in group B. Differences were statistically significant. CONCLUSION: Our findings are supportive of PD as an effective, cheap and easy-to-apply method for the reduction (a) of the time with drains in situ after MRM, (b) of the number of patients developing seromas and (c) of the seroma aspirations. This can potentially reduce further complications, needed medical care and cut expenditure. PMID- 18404790 TI - Association between bone scintigraphy and serum levels of procollagen (I) and PSA in the detection of bone disease in prostate cancer patients. AB - PURPOSE: To evaluate the clinical usefulness of serum procollagen I carboxyterminal propeptide (PICP) and prostate specific antigen (PSA) in relation to bone scan results in Greek patients with prostate cancer (PC). PATIENTS AND METHODS: 108 patients (mean age 58+/-4.3 years; range 42-81) with PC and 52 healthy blood donors as control group were examined for serum PICP and PSA levels. The diagnosis of PC was confirmed histologically. Bone metastases were diagnosed in 68 of the patients with the use of (99m)Tc-MDP bone scan, while 40 patients had no bone metastases. During the one year follow-up new PICP and PSA measurements were obtained along with a new bone scan for all groups studied. RESULTS: The levels of serum PICP and PSA were significantly higher in patients with PC and bone metastases in comparison to patients with no bone metastases. The sensitivity and specificity of the combination of PICP and PSA were 78% and 96%, respectively. CONCLUSION: PICP could be useful for diagnosing early bone metastases of prostate adenocarcinoma and in combination with PSA and bone scan can be an additional tool in the follow-up of patients with PC. PMID- 18404791 TI - Comparison of the 3D-conformal, helical tomotherapy and multileaf collimators based intensity modulated radiotherapy modalities using radiobiological measures. AB - PURPOSE: Intensity modulated radiotherapy (IMRT) using multileaf collimators (MLC) and helical tomotherapy (HT) have become increasingly popular over the past few years. However, their clinical efficacy and effectiveness continue to be investigated. In order to provide a more thorough evaluation and comparison of treatment plans, the utilization of the biologically effective uniform dose (D) together with the complication-free tumor control probability (P(+)) are examined. MATERIALS AND METHODS: In this study, a typical case of lung cancer was investigated by developing a 3D conformal treatment plan, a linac MLC-based step and-shoot IMRT plan and a HT plan. The 3 different treatment plans were compared based on radiobiological measures by using the P(+) index and the D concept as the common prescription point of the plans and plotting the tissue response probabilities vs. D for a range of prescription doses. RESULTS: The applied plan evaluation method showed that in this lung cancer case the MLC-based IMRT plan was best over the clinically useful dose prescription range. The 3D-conformal, MLC-based IMRT and HT treatment plans gave a P(+) of 55.4%, 72.9% and 66.9%, for a D to the internal target volume (ITV) of 57.0 Gy, 66.9 Gy and 64.0 Gy, respectively. CONCLUSION: In comparison to 3D conformal radiotherapy, both MLC based-IMRT and HT can better encompass the often large ITV required while minimizing the volume of the organs at risk receiving high dose. Taking into account the dose-response relations of the irradiated tumors and normal tissues, a radiobiological treatment plan evaluation can be performed, which may provide a closer association of the delivered treatment with the clinical outcome. PMID- 18404792 TI - Nitric oxide and the collagenous protein biosynthesis of irradiated chick chorioallantoic membrane. AB - PURPOSE: To study the effect of irradiation doses (2 Gy and 5 Gy) on the rate of collagenous protein biosynthesis (CPB) of chick embryo chorioallantoic membrane (CAM) and to investigate the possible role of the nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME), its inactive enantiomer D NAME and the NO donor sodium nitroprusside (SNP) in this effect. MATERIALS AND METHODS: The originally established CAM angiogenesis model was used with few modifications described earlier. The CAM areas were irradiated on the 10th or the 14th day of embryo development with a single dose of 2 or 5 Gy and the CPB was determined 6 h later. L-NAME, D-NAME and SNP were added to the CAM after the irradiation. RESULTS: The experiments didn't show any significant differences in almost all of the treatment courses. In the 10-day CAM preparations the irradiation not significantly (NS) inhibited the CAM CPB, whereas L-NAME abolished this effect only in the case of 2 Gy dose (NS). The addition of SNP or D-NAME in the post-irradiated 10-day CAM exerted radiosensitization that was significant only in the case of the combination of D-NAME with the 5 Gy dose. There was no significant effect on the respective treatments of the 14-day CAM. The CPB of the 14-day CAM was significantly lower with regard to the values of the 10-day CAM. CONCLUSION: The biochemical evaluation of the CAM (CPB method) seems to be not suitable for radiobiological studies. Nevertheless, the implication of NO in the X-ray induced antiangiogenicity cannot be excluded. PMID- 18404793 TI - Neuron specific enolase tissue expression as a prognostic factor in advanced non small cell lung cancer. AB - PURPOSE: To determine the frequency of neuron specific enolase (NSE) tissue expression and its possible influence on survival of patients treated for advanced non small cell lung cancer (NSCLC). PATIENTS AND METHODS: We studied 158 patients with histological diagnosis of NSCLC (stage III/IV). Combined chemoradiotherapy was used in stage III disease (without pleural effusion), whereas chemotherapy only was used in stage III (with pleural effusion) as well as in stage IV disease. Immunohistochemical analysis of NSE expression was determined using antibodies to NSE (DAKO, Den). One- and 2-year overall survival were assessed. RESULTS: 45 (28.5%) patients had positive NSE expression. The most frequent NSE expression was seen in 6/9 (66.7%) patients with large cell carcinoma and in 23/50 (46%) with adenocarcinoma. One- and 2-year survival rates were 70% and 30% in the group of patients with positive NSE expression compared to 26% and 3% in the group with negative NSE expression (p=0.000). The median survival time was 16.4 vs. 11.4 months in the groups of patients with and without NSE expression, respectively (p < 0.001). One-and 2-year survival rate was higher in NSCLC patients with more than 50% of NSE positive cancer cells (p=0.0004 and 0.000, respectively). CONCLUSION: A total of 28.5% of advanced NSCLC patients had positive NSE expression. Median 1- and 2-year survival time was significantly longer in patients with positive NSE expression. PMID- 18404794 TI - The histological outcome of glandular dyskaryosis--AGUS--reported in Papanicolaou smears. AB - PURPOSE: The objective of this study was to investigate the prognostic significance of glandular dyskaryosis/AGUS, reported in Papanicolaou (Pap) smears. MATERIALS AND METHODS: During a 4-year period 26,408 smears were assessed at the Department of Cytology of our hospital. Thirty (0.11%) smears were reported as having glandular dyskaryosis. The studied material was taken by colposcopy, fractional curettage and/or cone biopsy. RESULTS: The final diagnosis included 2 cases of invasive cervical carcinoma (1 squamous cell and 1 adenocarcinoma), 1 case with ovarian cancer, 8 cases with high grade squamous intraepithelial lesion (HGSIL) and 10 cases with other nonmalignant pathology (polyps, hyperplasia) of endometrial origin. These findings reflect a 36.7% positive predictive value for significant squamous and glandular pathology. CONCLUSION: Patients with glandular dyskaryosis require further evaluation because it may hide serious pathology from all internal genital organs. Colposcopy in combination with fractional curettage and/or cone biopsy are proposed as the appropriate diagnostic tools in women with such cytological abnormality. PMID- 18404795 TI - Two-dimensional electrophoresis and immunohistochemical study of calreticulin in colorectal adenocarcinoma and mirror biopsies. AB - PURPOSE: The purpose of this study was to analyse the polypeptide patterns of colorectal adenocarcinomas and mirror biopsies and to investigate the expression of calreticulin and the relationship of this chaperon to colon cancer. MATERIALS AND METHODS: The investigation was carried out on 21 adenocarcinomas and 21 mirror biopsies using high-resolution two-dimensional (2D) electrophoresis and immunohistochemical PAP method. RESULTS: 2D electrophoresis revealed several polypeptide patterns that were shown to be upregulated in colorectal adenocarcinomas compared to their mirror biopsies. One polypeptide spot being upregulated in colorectal adenocarcinoma, turned out to be calreticulin. The overexpression of calreticulin was confirmed by further examination in immunohistochemical level. CONCLUSION: Calreticulin was found overexpressed in colon cancer tissues as compared to the corresponding mirror biopsy tissues. The overexpression was particularly intense to high-malignancy tissues and particularly in the poorly differentiated regions of the tissue. Calreticulin showed a direct relationship to the disease stage, a fact strongly indicating that the functional role of calreticulin is directly associated with tumor growth and metastasis. PMID- 18404796 TI - A woman with synchronous cervical, vaginal and laryngeal squamous cell carcinomas and positive human papillomavirus type 16; case presentation with literature review. AB - During the last 25 years the development of cervical cancer and cancer of the anorectum has been related to human papillomavirus (HPV) infection. HPV infection is found in up to 99.7% of all cervical cancers. Head and neck squamous cell carcinomas (HNSCC) have also been related to HPV infection. In this communication we present a 67-year-old female patient, nonsmoker, diagnosed with 3 synchronous cancers--squamous cell carcinoma of the uterine cervix, vagina and larynx. The patient was found positive for immunoglobulin HPV 16 L1 using ELISA method. We believe that in the presented case, the high risk HPV subtype 16 was implicated in the etiology of laryngeal, cervical and vaginal cancers. We also think that HPV infection and related cancers could be prevented effectively by immunization. PMID- 18404797 TI - A patient with diffuse large B-cell non-Hodgkin's lymphoma and AA type amyloidosis. AB - AA-type amyloidosis is a consequence of a long-standing systemic inflammation and is not associated with a monoclonal protein or clonal bone marrow plasma cells. Proinflammatory cytokines such as interleukin (IL)1, IL-6, and tumor necrosis factor (TNF) stimulate the synthesis of serum amyloid A during inflammation. Although the association of non-Hodgkin's lymphoma (NHL) with AL-type amyloidosis is well known and patients with Hodgkin's lymphoma with AA amyloidosis have been described, AA-type amyloidosis with NHL is extremely infrequent. We report a case of amyloidosis associated with NHL that subsided during R-CHOP chemotherapy. PMID- 18404798 TI - Fibrous tumor of the lung: diagnostic and therapeutic considerations apropos of two cases. AB - Solitary fibrous tumor (SFT) of the pleura and the lung is an uncommon spindle cell neoplasm arising from the visceral pleura in the majority of the cases. Current diagnostic and therapeutic considerations are discussed apropos of 2 recent cases. 1st case: A 46-year-old male, heavy smoker, presented with nonspecific complaints and a mass in the left posterior mediastinum at the level of the 5th thoracic vertebra. Computed tomography (CT) and magnetic resonance imaging (MRI) of the chest confirmed the position of the mass in proximity to the thoracic wall at that level. Through a left posterolateral thoracotomy, a 4 x 5 x 2 cm mass, mushroom-shaped, stalky, completely covered by the lung parenchyma, was resected. Histopathological examination revealed a SFT of the lung. 2nd case: A 54-year-old woman presented with symptoms of respiratory distress and persistent hypoglycaemia. CT scan of the chest showed a huge mass within the right hemithorax, compressing the mediastinum and the contralateral lung. The mass was initially detected on chest radiography and CT scan 3 years before resection. Fine needle biopsy (FNB) results were inconclusive and resection of the mass through a right thoracotomy established the diagnosis of malignant SFT. Resection of the mass resulted in recovery of respiratory function and the paraneoplastic hypoglycaemia. Radiologic features and inconclusive fine needle aspiration (FNA)/FNB results make preoperative diagnosis of SFTs of the pleura and lung difficult. Diagnosis of SFT will be established with certainty after surgery. Resection with clear margins is the main important prognostic factor. PMID- 18404799 TI - Preoperative diagnosis of jejunal adenocarcinoma--a case report. AB - We report herein the case of a preoperatively diagnosed small bowel adenocarcinoma. A 57-year-old man was hospitalized twice for a month in the Gastroenterology Clinic due to complaints lasting a year before hospitalization and consisting of heaviness and spastic pain in the left upper abdomen. None of the numerous methods used in this case (fibrogastroduodenoscopy, ileocolonoscopy, barium series and CT) could reveal a tumor. Ultrasound (US) examination showed a polycyclic formation in the left hypochondrium with enlarged regional lymph nodes. An US-guided fine (20 gauge) needle aspiration (FNA) cytology of the mass was performed and showed moderately differentiated adenocarcinoma of intestinal origin. Thanks to Doppler US guidance during FNA a massive bleeding from an identified arterial vessel with high systolic speed was avoided. On surgical exploration a jejunal tumor of high consistency was found, located 15 cm distantly from the Treitz ligament, infiltrating up to 2/3 of the bowel circumference and partly obstructing its lumen. The histological evaluation of the resected material showed highly to moderately differentiated adenocarcinoma with 5 regional lymph node metastases. The combination of US with Doppler and FNA established preoperatively the malignant small bowel disease. PMID- 18404800 TI - Hibernoma: a benign lipomatous tumor mimicking liposarcoma. AB - Hibernoma is a rare, benign, slow-growing soft tissue tumor. It was named after its resemblance to the brown fat found in hibernating animals. Due to its rich vascularity and magnetic resonance appearance, this tumor may mimic a liposarcoma before tissue diagnosis. Complete resection with meticulous hemostasis is enough for treatment. Malignant potential has not been demonstrated. Herein we present a case of hibernoma arising from the left groin in a young man. The clinical presentation, radiographic and histopathologic features of this rare benign soft tissue tumor are presented. This tumor is clinically important because it is indistinguishable from malignant lesions. PMID- 18404801 TI - Term and role of tumor centres in oncology healthcare in Germany. AB - Breast cancer is the most common malignant disease among women in Germany. There are more than 50,000 new cases of breast cancer diagnosed each year in the country. As breast cancer is a complex disease it necessitates an interdisciplinary treatment, therefore it is very likely that patients with breast cancer will benefit from treatment in special centers, either a "breast cancer centre" or "tumor centre". In recent years certifying programs for the above mentioned centers have been introduced, which guarantee standards of the quality of treatment for breast cancer all over the country. In this article we report on the development and role of tumor centers in oncology healthcare in Germany, which nowadays have a very good international reputation. Moreover, they may be a good example to other countries to found similar centers in order to improve standards of the management in oncology healthcare. PMID- 18404803 TI - Cutaneous diphtheria masquerading as a sexually transmitted disease. AB - A 41 year-old, sexually promiscuous, married male, an agricultural laborer by occupation, presented to our sexually transmitted diseases (STD) clinic with multiple ulcers over the scrotum and genitalia of 20 days' duration. Bacterial culture from swabs taken from the genital ulcer, grew organisms morphologically and biochemically characteristic of Corynebacterium diphtheriae. He made a complete and uneventful recovery after two weeks of therapy with antidiphtheria serum and crystalline penicillin. This case brings into light this hitherto unreported presentation of wound diphtheria mimicking a sexually transmitted genital ulcer disease and thus, underlines the importance of considering diphtheria as differential in atypical, long-standing genital ulcers. PMID- 18404802 TI - The great oncologic surgery of the eminent Romanian anatomist and surgeon Thomas Jonnesco (1860-1929). AB - Thomas Jonnesco was an eminent Romanian surgeon and anatomist. Politician and patriot, he was founder of the modern Romanian experimental surgery and initiator of general rachianaesthesia. Among his scientific works we underline his contribution to the field of oncology since he established new original surgical methods for the treatment of cancer. PMID- 18404804 TI - Patch test in Behcet's disease. PMID- 18404806 TI - Current awareness in NMR in biomedicine. PMID- 18404805 TI - Vesicles on the tongue. PMID- 18404807 TI - Influenza in Canada: 2006-2007 season. PMID- 18404808 TI - The Mosaic of Autoimmunity. Proceedings of a meeting in honor of Professor Shoenfeld in Tel Aviv, Israel, February 10-11, 2008. PMID- 18404810 TI - Proceedings of the European Society for Clinical Virology symposium on respiratory viruses, 26-27 May 2007, Lyon, France. PMID- 18404809 TI - Investigation of the second wave (phase 2) of severe acute respiratory syndrome (SARS) in Toronto, Canada. What happened? AB - BACKGROUND: Severe acute respiratory syndrome (SARS) was introduced to Toronto on 23 February, 2003. The outbreak was believed to be over in early May after two incubation periods had passed with no newly identified cases. However, on 20 May, 2003 a cluster of respiratory illnesses occurred in a rehabilitation facility in Toronto. These cases were later identified as SARS but not until a community hospital in which one of these cases was previously hospitalized (Hospital X) had already experienced nosocomial transmission. This report describes the outbreak investigation of nosocomial transmission of SARS at Hospital X. METHODS: An investigation of 90 probable and suspect cases of SARS associated with a hospital outbreak was performed. The investigation included death record reviews, chart reviews, case finding and contact tracing. Nursing cohorts who worked on the specific ward in which nosocomial transmission occurred had work-shift and patient-assignment records reviewed to determine source of infection. RESULTS: The greatest number of cases occurred within hospital employees (42.5%) with an average age of 51 years and 69% being female. The mean incubation period for one time exposure patients was 6.3 days (range 4 to 10 days). Twelve (13.8%) of the cases died. Five of seven nurses who cared for a specific SARS patient during this period acquired SARS. Twelve of 17 (70.6%) nursing staff who developed SARS worked with another nursing staff who was symptomatic for SARS. Staff members who worked the evening shift or the morning shift and therefore likely attended a nursing appreciation breakfast were five times more likely to acquire SARS than those who did not attend. INTERPRETATION: What was believed to be the end of the Toronto SARS outbreak led the Provincial Operations Centre (POC) to issue a directive allowing a more relaxed use of infection-control precautions during the beginning of Phase 2 of the outbreak. These relaxations of precautions were temporally associated with the nosocomial transmission of SARS to hospital staff, other patients and visitors at Hospital X. As a result of this outbreak significant changes have been made with respect to infection-control practices within Canada. PMID- 18404811 TI - Autoimmunity: From the mosaic to the kaleidoscope. Proceedings of a symposium in honor of Professor Shoenfeld in Tel Aviv, Israel, February 10-11, 2008. PMID- 18404812 TI - [Back and radiology II]. PMID- 18404813 TI - Colorectal Cancer: Biology, Diagnosis and Therapy, Varese, Italy, September 26 27, 2007. Conference proceedings. PMID- 18404815 TI - Proceedings of the 2007 International Berry Health Benefits Symposium, June 11 12, 2007, Corvallis, Oregon, USA. PMID- 18404814 TI - The seven deadly sins of business development. PMID- 18404816 TI - Cellular and molecular engineering. Issue celebrating the 80th birthday of Professor Harry Goldsmith. PMID- 18404817 TI - Proceedings of the Asia Pacific Bioinformatics Network (APBioNet) 6th International Conference on Bioinformatics (InCoB207), Hong Kong, China, 27-30 August 2007. PMID- 18404818 TI - [Appearance and diagnosis of intracavitary uterine pathologies]. PMID- 18404819 TI - [Diagnosis of uterine malformation]. PMID- 18404820 TI - [Semiology of ovarian cysts and tumors]. PMID- 18404821 TI - [Uterine adenomyosis or endometriosis]. PMID- 18404822 TI - [Imaging of fibromas]. PMID- 18404823 TI - [The ovary: folliculogenesis and ovulation disorders]. PMID- 18404824 TI - [Plasma creatinine and cystatin C ratio is useful for discriminate diagnosis of postrenal renal failure]. AB - Acute renal failure (ARF) is an acute loss of kidney function that occurs over days to weeks and results in an inability to appropriately excrete nitrogenous wastes and creatinine (Cre). ARF is diagnosed by elevations of blood urea nitrogen and serum Cre level, which is classified as prerenal, intrinsic and postrenal according to their mechanisms. However, discriminate diagnosis of these types by blood biochemistry findings is difficult. Recently, cystatin C (Cys-C), a basic protein having isoelectric point 9.3 with a molecular weight of 13.3 kDa, is freely filtered at the level of the glomerulus and virtually all is reabsorbed and metabolized by the proximal tubular cells. Therefore, assuming constant cellular production, serum Cys-C level has the potential to be an excellent surrogate marker of glomerular filtration rate. Because Cre is electrically charged neutrally, there is a possibility that the permeation of Cys-C, which is positively charged, is diffluent from that of Cre through glomerular basement membrane due to the type of the renal failure. We determined blood concentrations of Cys-C and Cre in a patients with prerenal renal failure (17 patients), intrinsic renal failure (232 patients) and postrenal renal failure (13 patients) as compared with healthy subjects (n = 771). We found that patients with postrenal renal failure displayed significantly elevated Cre/Cys-C ratio (mean +/ standard deviation) (8.3 +/- 8.0, p < 0.001) as compared with healthy subjects (1.1 +/- 0.2), prerenal (0.6 +/- 0.2) and intrinsic (1.6 +/- 0.5). These findings suggest that measurement of Cys-C concentration and Cre/Cys-C ratio may be useful for the discriminate diagnosis of postrenal renal failure. PMID- 18404825 TI - [Advance in technical method and treatment for viral hepatitis]. AB - The dominant treatment used currently for hepatitis B is administration of interferon for 6 months or long-term administration of nucleic-acid analogs. However, since the response rate to interferon treatment is approximately only 30%, nucleic acid analogs are used primarily for the treatment of hepatitis B in our country. Among the nucleic-acid analogs, entecavir is associated with the lowest rate of emergence of mutants (approximately 3% for 3 years). On the other hand, combination therapy with interferon plus ribavirin is predominantly used in the treatment of chronic hepatitis C, and the response rate is approximately 50% in cases with genotype 1b infection and high viral loads and 90% in cases with genotype 2a/2b and high viral loads. Moreover, because interferon monotherapy has been shown to be associated with a 70% response rate in subjects with low viral loads regardless of the genotype, combined Peg-IFN plus ribavirin therapy and interferon monotherapy are mainly used for hepatitis C patients with high and low viral loads, respectively. beta-interferon is mainly used for preventing the development of cancer in patients with compensated cirrhosis, in addition to its use in cirrhosis patients with genotype 1b infection and high viral loads. PMID- 18404826 TI - [Diagnosis and treatment in systemic amyloidosis]. AB - Systemic amyloidosis is characterized by the involvement of multiple organs and the presence of an amyloid precursor protein in serum. This disorder is classified into four major forms: immunoglobulin light chain-derived (AL), reactive AA, dialysis-related (beta2M) and hereditary transthyretin (ATTR) type. Heart, kidney, gastrointestinal tract and peripheral nerves are commonly affected by amyloid deposition in systemic amyloidosis and histopathological demonstration of amyloid deposits on any of affected organs is the first step leading to the diagnosis of this disease. Immunohistochemical analysis of amyloid protein on tissue amyloid deposits is necessary to make classification of the disease and DNA testing is also useful in a hereditary form. Amyloidosis had been considered to be an incurable disease but during the past one decade several therapeutic approaches have been employed for the amyloidosis patients with diverse pathogenetic backgrounds: intravenous large dose of melphalan accompanied by autologous peripheral blood stem cell transplantation for AL amyloidosis and liver transplantation for hereditary ATTR type amyloidosis. As a result some amyloidosis patients have been rescued and are now enjoying their own social lives. It is likely that recent advance on the research of amyloidosis has changed the concept of this disease. PMID- 18404827 TI - [Conformational change, pore formation and neurotoxicity of amyloidogenic proteins]. AB - Conformational changes of Alzheimer's beta-amyloid protein (AbetaP) enhance its neurotoxicity, and finally lead to Alzheimer's pathogenesis. Recent studies have suggested that a common mechanism is based on the diverse diseases termed "conformational diseases", including other neurodegenerative diseases such as prion diseases, Parkinson's disease, and triplet-repeat disease. These diseases share similarity in the formation of beta-sheet containing amyloid fibrils by disease-related proteins including prion protein, alpha-synuclein, polyglutamine, and the introduction of apoptotic degeneration. We have investigated the conformational changes of AbetaP and its neurotoxicity and found that trace metals including aluminum enhanced the aggregation of AbetaP. Although the molecular mechanism of neurodegeneration induced by these conformational disease related proteins remains elusive, these proteins have the ability to directly incorporate into membranes directly and to form calcium-permeable ion channels. Our and other numerous studies have revealed that AbetaP caused the abnormal elevation of intracellular calcium levels. We review here the current understanding of the pathology of the conformational diseases based on the hypothesis that the disruption of calcium homeostasis through amyloid channels may be the molecular basis of the neurotoxicity of AbetaP and other disease related proteins. PMID- 18404828 TI - Patient safety alert. Initiative gives families access to rapid response team. PMID- 18404829 TI - Placement of posterior resin restorations should be limited to dentists. PMID- 18404830 TI - Ovulation induction and pregnancy in a woman with premature menopause following gonadotropin suppression with the gonadotropin releasing hormone antagonist, cetrorelix--a case report. AB - OBJECTIVE: To determine if ovulation and pregnancy could be achieved in a case of amenorrhea, estrogen deficiency, and markedly elevated serum follicle stimulating hormone (FSH) through reduction of the serum FSH by a gonadotropin releasing hormone antagonist. METHODS: A 37-year-old woman with hypergonadotropic secondary amenorrhea related to two courses of chemotherapy with alkylating agents and abdominal radiation therapy (Hodgkin's disease and breast cancer) was treated with cetrorelix in an attempt to induce ovulation by lowering elevated serum FSH and hopefully restore sensitivity of the few remaining follicles by restoring down-regulated FSH receptors. She was monitored with serum estradiol (E2), FSH, luteinizing hormone (LH), progesterone (P) levels and sonography. RESULTS: As the serum FSH dropped the serum E2 rose and peaked at 200 pg/ml after ten days of cetrotide. She conceived in that cycle. A viable ongoing pregnancy with appropriate ultrasound findings was demonstrated 40 days from conception. CONCLUSION: This is the first case description of successful ovulation and pregnancy following induction of ovulation with the GnRH antagonist cetrorelix. The possibility exists that the ovulation was spontaneous but it seems unlikely. It has been estimated that the chance of spontaneous ovulation and pregnancy in cases of premature ovarian failure is 1:9,200. PMID- 18404831 TI - Elimination of yaws in India. PMID- 18404832 TI - [Progress in research and clinical practice related to olfactory sense- regeneration medicine for olfactory nerve cells]. PMID- 18404833 TI - Most precious cargo: child restraints continue to save young lives, but gaps persist in many state laws. PMID- 18404834 TI - [High quality of life and long survival need to be considered in surgical treatment for oesophageal carcinoma]. PMID- 18404835 TI - [Reconstruction of esophagus with gastric tube: view from anatomical physiology]. PMID- 18404836 TI - [Medical treatment of peripheral artery diseases]. PMID- 18404837 TI - [Epidemiology and prognosis of peripheral artery diseases]. PMID- 18404838 TI - [Progression and attrition of arteriosclerosis]. PMID- 18404839 TI - [QOL in peripheral artery diseases--from studies to produce results to individual management]. PMID- 18404840 TI - [New diagnostic and therapeutic guideline TASC-II for peripheral arterial diseases]. PMID- 18404841 TI - [Bedside diagnosis of peripheral arterial diseases (from medical history taking to physical examination)]. PMID- 18404842 TI - [Vascular laboratory and diagnosis of peripheral arterial diseases]. PMID- 18404843 TI - [Functional diagnosis of peripheral artery diseases]. PMID- 18404844 TI - [Ultrasonic diagnosis of peripheral arterial diseases]. PMID- 18404845 TI - [Multislice CT of peripheral arterial diseases]. PMID- 18404846 TI - [Internal treatment (drug and exercise therapies) of peripheral arterial diseases]. PMID- 18404847 TI - [Endovascular treatment of peripheral arterial diseases]. PMID- 18404848 TI - [Surgical therapy of peripheral arterial diseases]. PMID- 18404849 TI - [Management of daily lives and guidance of patients with peripheral arterial diseases]. PMID- 18404850 TI - [Management of critical limb ischemia]. PMID- 18404851 TI - [Angiogenic therapy for critical limb ischemia]. PMID- 18404852 TI - [Rising incidence of peripheral arterial diseases (PAD): its diagnosis and treatment--to be thoroughly familiar with diagnostic sequences and others matters related to PAD. Discussion]. PMID- 18404853 TI - [Frontier of Q fever management]. PMID- 18404854 TI - [Current therapy of hypertension]. PMID- 18404855 TI - [Auto-inflammatory syndrome]. PMID- 18404856 TI - [Case of microscopic polyangiitis associated with reversible posterior leukoencephalopathy syndrome (RPLS)]. PMID- 18404857 TI - [Interpretation of the appropriate guideline for elective coronary angioplasty for the cases with stable angina pectoris]. PMID- 18404858 TI - ["Minimum knowledge" needed by primary care physicians. Management of patients with upper abdominal symptoms by primary care physicians]. PMID- 18404859 TI - [The internist as a generalist]. PMID- 18404860 TI - [Zebra Cards J-(1)]. PMID- 18404861 TI - Missouri report reveals enrollment, cost trends. PMID- 18404862 TI - Leverage market competition for government contracts. PMID- 18404863 TI - Maine program examines performance-based initiatives. PMID- 18404864 TI - Emerging topical antimicrobial options for mild-to-moderate acne: a review of the clinical evidence. AB - Topical antimicrobial agents are commonly used as first-line agents for the management of mild to moderate acne vulgaris. Agents with antibacterial activity (eg, benzoyl peroxide and clindamycin), reduce counts of Propionibacterium acnes, while also decreasing the numbers of inflammatory and noninflammatory lesions. In order to optimize efficacy, topical antibiotics are most commonly used in combination with benzoyl peroxide, an approach which also reduces the emergence of strains of P acnes that are less sensitive to antibiotics. Other topical antiacne options include retinoids (with or without an antimicrobial agent), sulfacetamide, sulfacetamide-sulfur, and azelaic acid. Certain formulations of benzoyl peroxide offer improved tolerability, while a new micronized form may enhance efficacy by optimizing follicular penetration. This wide array of options allows dermatologists to tailor topical treatments to the needs of individual patients. PMID- 18404865 TI - New developments in topical antimicrobial therapy for acne. AB - Benzoyl peroxide (BPO) has been an important component of the acne treatment armamentarium for decades. A number of characteristics contribute to its success in acne, including its lipophilicity, potent antibacterial activity, and ability to suppress antibiotic-resistant strains of Propionibacterium acnes. With some commercially available products in which BPO comprises crystals in suspension, aggregation of BPO can occur on the skin surface in large clumps, resulting in poor penetration of sebaceous follicles. In addition, the poor water solubility of BPO, coupled with its chemical instability in other solvents, presents challenges with respect to formulating topical products with optimal bioavailability, stability, and tolerability. Several products and vehicle systems have evolved in attempts to improve on the benefits of BPO in treating acne. A new acne treatment has been developed that contains 5% solubilized BPO consisting of small-size particles. This new product exhibits enhanced follicular penetration of BPO, improved P acnes kill rates and clinical efficacy, and good consumer acceptance relative to existing formulations. PMID- 18404866 TI - The role of elastin and collagen in cutaneous aging: intrinsic aging versus photoexposure. AB - Cutaneous aging occurs through 2 biologically distinct processes: intrinsic and extrinsic aging. The first is a naturally occurring process that results from slow tissue degeneration. In human dermis, intrinsic aging is characterized by 3 features: atrophy of the dermis due to loss of collagen, degeneration in the elastic fiber network, and loss of hydration. In contrast to intrinsic aging, extrinsic aging is due to environmental factors. Since ultraviolet (UV) exposure is the principal cause of extrinsic aging, it is often referred to as photoaging. At the microscopic level, the distinguishing feature of photoaging is a massive accumulation of elastotic material in the upper and middle dermis, a process termed solar elastosis. Using recombinant DNA technology, it has become possible to demonstrate that UV radiation can activate the human elastin promoter. This provides a mechanism for enhanced elastin biosynthesis, which contributes to the clinical and morphologic changes observed in photoaged skin. PMID- 18404868 TI - Complementary effects of topical antiaging treatments in conjunction with aesthetic procedures. AB - The dermatologist has a variety of tools for improving the appearance of aging skin. These include injectable botulinum toxins and dermal fillers, laser treatments, chemical peels, and various topical agents, including cosmeceuticals. A combined approach using more than one facial rejuvenation tool is considered ideal for many patients, as it targets the various areas of the face and numerous dynamic and static changes associated with aging. A topical cosmeceutical regimen, such as one containing tretinoin and hydroquinone, can enhance the effects of facial rejuvenation procedures and encourage patients to adopt a daily cleansing and rejuvenation regimen that they can continue after the procedure to help maintain the aesthetic effects. PMID- 18404867 TI - Cosmeceuticals for the attenuation of extrinsic and intrinsic dermal aging. AB - Since the term "cosmeceutical" was coined over 2 decades ago, the number of products in this category that claim to combat dermal aging has grown dramatically. Topical retinoids remain the mainstay for treating photoaging given their proven efficacy in both clinical and histologic outcomes. In addition to retinoids, many other cosmeceutical agents are now available. The proliferation of products can cause confusion among consumers, who often ask their dermatologist for advice as to which antiaging products they should choose. Ideally, the antiaging claims of cosmeceutical formulations and their components should be demonstrated in controlled clinical trials. In order to provide appropriate recommendations to their patients, dermatologists must become familiar with the available data on currently marketed products and gain experience with antiaging regimens. This review discusses the efficacy of a number of currently marketed drug products with proven photoaging benefits and cosmeceutical products that claim similar benefits. Among the agents discussed are single-entity and combination products containing hydroquinones, retinoids, topical antioxidants, and minerals. PMID- 18404869 TI - Optimal control of psychological processes: a new computational paradigm. AB - In this paper, we use general mathematical-statistical theorems to prove that developmental processes must be studied at the intra-individual level. We demonstrate how to model intra-individual variation using single-participant time series analysis with time-varying parameters. We use advanced signal analysis techniques based on nonlinear state-space modeling to present simulation results obtained with a new Maximum Likelihood technique based on Extended Kalman Filtering with Iteration and Smoothing (EKFIS) embedded in an Expectation Maximization (EM) loop. After showing how EKFIS results yield state-space models with time-varying parameters, we then couple EKFIS to recursive optimal control techniques to produce a receding horizon feedback-feedforward controller. In this way, we obtain a flexible on-line computational paradigm with which we can optimally control observed behavioral processes for an individual person in real time. We will present optimal control techniques using simulated data and outline preliminary applications to real time patient-specific treatment of type I diabetic patients and asthma patients. PMID- 18404870 TI - Functional configuraI frequency analysis: explaining types and antitypes. AB - In this article, we ask how types and antitypes in Configural Frequency Analysis (CFA) can be explained at the level of effects in log-linear models. Two strategies are proposed for the search for the effects that cause types and antitypes to emerge. The first strategy is ascending and inclusive. Beginning with the CFA base model, effects are systematically added until the most parsimonious model is found that meets the following two conditions. The first condition is that all types and antitypes disappear. The second condition is that no new types and antitypes emerge. The second strategy is descending and exclusive. Beginning with the saturated model, effects are systematically excluded until (1) the effects are identified that would make all types and antitypes disappear, and (2) no new types and antitypes emerge. A first comparison of the two models suggests that the ascending, inclusive strategy yields more parsimonious models. However, results from the descending, exclusive strategy have a more clear-cut interpretation. Data examples are given that describe the development of severe domestic violence. PMID- 18404871 TI - [Bases and principles of a qualitative approach in the phenomeno-structural analysis applied to Rorschach's method]. AB - In the days when a first assessment was becoming necessary, twenty years after the first edition of Rorschach's Psychodiagnostic, which was being edited for the 4th time, and while he was worried about the readjustment of the scientific bases of this experiment, his friend and colleague Doctor Walter Morgenthaler was feeling the need to warn against the use, teaching and interpreting of this method of approaching personality in a too mechanical way, saying: "For some scientific problems and statistical research the simple quotation can be enough or at least the main thing, but for the very diagnosis of the personality, the mechanical side, especially the mathematical accuracy, although necessary, is nothing but a base. Many errors in the use of Rorschach's method are coming from the fact that the mechanical framework is taken as the whole result." (Psychodiagnostics, P. 254) Discovering and applying this method at around the same time, Francoise Minkowska is going to feel as well the need for its qualitative access, putting accents or suggesting proceedings in theoretical patterns, procedures and interpretation of results which will influence an overall direction close to Morgenthaler's one in this matter: "The psychogram, through its quantitative aspect, contains something static and deprives us of essential data, namely the way in which the test is going on, a process which shows us continuous comings and goings depending on various elements belonging to the person tested, to the climate of each plate, and even to the contact with the person of the psychologist" (Rorschach. Towards the research of the world of forms, P. 226). Thus, for her, a deepened qualitative analysis of the language can lie neither on word frequencies nor occurrences, nor on a metric index, but has to be necessarily combined with a meticulous observation of the patient's attitudes and contexts of expression, to claim to restore the sharp, the vivid, core, dynamism and structure of one's personality characteristics. This paper tries to work out the principles and the authentically clinical bases of this process, not so much as a rupture than in continuity with the spirit and practice of Hermann Rorschach. PMID- 18404872 TI - The analysis of farewell letters of suicidal persons. AB - The aim of this article is to present some possibility of analysis of the letters written by persons committing suicide. A methodology of this type of analysis could be necessary for criminal investigations and forensic document expertise. The authors of those letters express their private and specific perception of the world, of people and themselves. These documents are very rich and plenty of intimate images. First of all the symptoms of presuicidal syndrome could be expressed in these letters. The author of this manuscript describes the suicidal syndrome characteristics that could possibly be observed in the farewell letters; then she attempts to present some proposals of analysis based on narrative theories and psycholinguistic research. According to these conceptions, an analysis of form and content could be conducted. Narrative psychology gives us the techniques of text study, such as lexical analysis, syntax analysis or cohesion analysis; these techniques could be useful in the field of investigation of farewell letters. PMID- 18404873 TI - [Extracting specific profiles in two clinical subgroups of patients suffering from addiction]. AB - According to research literature, people suffering from addiction to alcohol or illegal drugs are presenting well defined troubles of identity. The aim of this study consists in showing structural similarities and differences between those two clinical subgroups, with the help of the Rorschach test which we have interpreted in an integrated quantitative and qualitative manner. We constructed a rating scale for the test of Rorschach, following the phenomenological and structural tradition of Mucchielli. This rating scale is considering as well the special phenomena as the global judgment of the experienced clinical psychologist. It is differing from the classical psychogramme by its holistic perspective, as it takes into account the interaction of different factors in a specific context. In an exploratory perspective we applied this rating scale to the Rorschach protocols of two small samples of people suffering from alcoholism and of people suffering from drug addiction. The patients belonging to both subgroups were in a state of abstinence at the period of the test. Thus their answers are reflecting an interaction of their basic personality and long term effects of the toxic products, but not the immediate effects of alcohol or illegal drugs. We have compared the answers of the two subgroups of dependent people with the help of non parametric statistics. Their specific profiles have been extracted with optimal scaling techniques. The results are discussed in relationship with structural psychopathology. PMID- 18404874 TI - [Extracting predictive factors of anorexia nervosa and bulimia nervosa: an cross cultural study]. AB - This study aims to explore, within the framework of theories of self-regulation, risk profiles for anorexia nervosa and bulimia nervosa, in two countries of different cultures: Poland and France The studied population consists of 106 women of Polish (from Silesia) and French (from the region around Metz) nationality. We examine a group of 30 Polish and 32 French anorexics as well as 30 Polish and 14 French bulimics. All the participants filled in the EDI by Garner and al. (1983), the SEI by Coopersmith (1984), the GIFS by Maes et al. (2000), the Brief COPE by Carver (1997), the SECS by Van Elderen et al. (1997), the HADS by Zigmond and Snaith (1983) and the IPC by Levenson (1981). Our results indicate that the profile of Polish anorexics closely resembles that of French anorexics. There are two independent variables which play an equally important role: the interiorisation of anger and conflicts in the goals of intellectual development (the importance and at the same time the difficulty of reaching these goals). The factors predicting bulimia nervosa are not the same in the two populations under study. PMID- 18404875 TI - [Non linear principal component analysis and clues for a differential approach of five groups experiencing social precariousness]. AB - We wished to develop an original way of taking care of people experiencing great social precariousness. Our purpose was to develop communication and relational skills, to stimulate expression of emotions and feelings, to bring out personal resources, to increase well-being, motivation and self-esteem, and thus favour rehabilitation. Our sample is composed of long-term unemployed people, of people benefiting from measures of integration into the working process, of people living in community homes, of drug addicts, and of drug-addicted female prisoners. Our research is based on an integrated quantitative and qualitative methodology, with rating scales for the artistic production and observational frames for items of verbal and non-verbal behaviour completing the psychometric questionnaires. It is an action research; we use art therapy, which is a common practice in the health sector, especially with subjects having problems expressing there feelings through words. We have carried out a non linear principal component analysis (PRINCALS) on the data of the projective test (Rotter), as well as a between groups comparison of the responses to the questionnaire on life satisfaction (FLZ), with the help of the Mann-Whitney test. It is from these comparisons that we are able to draw out a few clues for differential treating strategies, depending on the inclusion into the five sub groups that we have followed. PMID- 18404876 TI - [Statistical analysis of renal tumours with infants]. AB - By using medical imaging a diagnosis for a patient is to be given. The shape of three-dimensional medical relevant objects in the human body is the basis for an accurate diagnosis. Based on the correct diagnosis, the appropriate therapy can be decided. In the following study the shape of renal tumours is explored by the mathematical method of shape analysis and by the test of Ziezold (1994). There are n=24 patients whose frontal view records are available. The possibility of classification can be presented only in an exploratory manner, due to the low case numbers of patients. PMID- 18404878 TI - [Study of the positive effect of work and nature on various aspects of child development at the preschool age]. AB - The goal of this study consists in showing the positive effects of contact with nature, linked to some well structured activities, on some developmental aspects with 4 to 6 years old children. In a semi-structured setting children belonging to the experimental group had the opportunity to meet and explore spontaneously nature with all its aspects and to discover natural phenomena linked to evolution and life, mirroring some analogous principles of their own development. Nature functioned as a free space, offering multiple possibilities to play and explore freely, aside from any kind of evaluation and consideration based on priority. With regard to the complexity of the theme and the great amount of possible interconnections, a holistic approach was appropriate. With the help of a quasi experimental design, we evaluated the data of two groups of children, analyzing not only their development, but also the relationships with their familiar environment and their involvement with nature. Thanks to three different procedural approaches, it was possible to show the differential progress of the experimental group in comparison with the control group, to select variables responsive to contact with nature and to bring out the positive link between good emotional contact with nature and general development. PMID- 18404877 TI - [Singing for preterm born infants music therapy in neonatology]. AB - The use of music as part of a stress reduction therapy has been applied both to premature infants and their parents in the Neonatal Reanimation Service. This aim of music therapy amounts to an attempt to help the premature infant regaining its physical and neurological balance, so important to its psychological and physical development, mainly by masking the sometimes excessive noise present in the intensive care unit and/or in the incubator. Studies have demonstrated the positive impact of music therapy on oxygen saturation, heartbeat, and on the general level of relaxation experienced by premature infants. In this project, the palliative technique used was that of live singing, directly to the infant, accompanied by a pentatonic harp. The aim was to improve the state of health, both physical and psychological, of a group of premature infants, whose gestation period varied between 23 and 36 weeks. The technique used was to apply what amounts to a protective cocoon of sounds to a premature infant in the neonatal unit, which measurably reduced the level of stress as indicated by the babies' increasingly relaxed demeanour and induced a measurable increase on the level of oxygen saturation and a reduction of heart rate. PMID- 18404879 TI - [Comparative study of two kinds of addictive behaviour; smoking and drug abuse. Interest of arts psychotherapy]. AB - The aim of the study consists in showing differences between addiction to tobacco, and to illegal drugs in terms of self-regulation skills, anxiety and depression. These two kinds of dependency are considered to take place on a continuum of addiction. Researches on emotional troubles in addictions have shown that there was a lower level of emotional awareness in patients with addictive behaviour, when compared with normal controls. As arts allow a better access to the emotional register, this kind of psychotherapy seems to be very interesting in cases of addiction. The comparison will be based on the HADS, with its two subscales of anxiety and depression, and on the SRSS, with the following subscales: avoiding distraction, control-regulation, planning, self-efficacy. From the results of our comparative study, we shall draw conclusions allowing us adapting our treatment by arts psychotherapy to different kinds of addictions. PMID- 18404880 TI - Analysing the relationship between traumatic biographical events and the current structural functioning of personality. AB - We present the general structure of a multi-annual research project. Our general expectancy concerns the possibilities of arts psychotherapy as a means of launching the blocked process of subjectivation with people suffering from exclusion, precarity and marginalization. The research project follows a complex research design with a sequential strategy, the first part consisting in an integrated psychosocial and clinical study using a mixed methodology. We constructed special rating scales for the analysis of the data of a semi structured biographical interview and also for the holistic interpretation of the Rotter Blank Sentences Test, separating the associations to sentences beginning with the third and first person. The correlations between two sets of variables (biographical interview and Rotter test) were computed for the total experimental group (N=206), and for clinical subgroups. We shall analyse the matrices of correlations (Spearman's Rho) with the help of optimal scaling procedures (OVERALS). The links between traumatic biographical events and responses to the 3rd, respectively 1st person items of the Rotter test are interpreted in terms of unconscious versus conscious psychological processes and allow us analysing the expression of defence mechanisms and coping strategies. The results of the study are discussed in the light of the recent traumatogenic hypothesis of borderline functioning. PMID- 18404881 TI - [Clinical characteristics of prostatic adenocarcinoma with ductal features]. AB - PURPOSE: To determine the incidence and prognosis of prostatic ductal adenocarcinoma. MATERIALS AND METHODS: From November 2002 to December 2005, we performed 122 radical prostatectomies and pelvic lymph node dissections. We further analyzed 64 cases after excluding 56 cases that had received neoadjuvant hormone therapy and two cases for which we were only able to perform limited follow-up examinations. We reviewed all of the surgical specimens and reclassified them according to the 2004 WHO classification system. We differentiated prostatic cases of ductal adenocarcinoma that were larger than 5 mm in diameter from cases of acinar adenocarcinomas. We then examined these two groups for the pathological stages of the neoplasms and the incidence of postoperative prostate-specific antigen (PSA) failure. Postoperative PSA failure was defined as a PSA value more than 0.2 ng/ml. RESULTS: We found eight cases (12%) of prostatic ductal adenocarcinoma among the 64 cases treated with radical prostatectomies. The mean age (+/- SD) of these patients was 65.3 (+/- 4.3) years old, and the mean PSA level (+/- SD) was 12.4 (+/- 5.4) ng/ml. Seven of the cases (11%) were mixed-type ductal adenocarcinomas, which contained acinar and ductal components. In addition, one case was identified as pure ductal adenocarcinoma. Seminal vesicle invasion was detected in four cases and lymph nodes metastases were identified in one case. During the follow-up period, four of the eight cases of ductal adenocarcinoma (50%) and twelve of the 56 cases of acinar adenocarcinoma (21%) showed postoperative PSA failure. The median follow-up period was 24 months (range: 12 to 48 months). CONCLUSIONS: We identified eight cases of ducal adenocarcinoma (12% of the examined cases), which suggests this disease is not as rare as previously reported. Compared to the cases of acinar adenocarcinoma, the cases of ductal adenocarcinoma were at a more advanced pathological stage and resulted in a higher rate of postoperative PSA failure. Therefore, we believe that patients that show even a limited degree of ductal adenocarcinoma should receive aggressive therapy. PMID- 18404882 TI - [Assessment of quality of life in women with pelvic organ prolapse: conditional translation and trial of P-QOL for use in Japan]. AB - INTRODUCTION: A purpose of the surgical treatment for pelvic organ prolapse (POP) is not only the anatomical restoration of the patients but also the improvement of their quality of life (QOL). We have no linguistic validated questionnaire in Japanese to assess it. Therefore we have translated 'prolapse quality of life questionnaire (P-QOL)' into Japanese and present it to the thirty POP patients at our institute for evaluating mental and physical condition before and three months after the operation. PATIENTS AND METHODS: We completed SF-36 and a questionnaire of patients' satisfaction on 30 patients besides P-QOL. SF-36 stands for Medical Outcome Study, Short Form 36 and is used for assessing overall QOL. We also completed pelvic organ prolapse quatification (POP-Q) to evaluate physical condition of the patients. RESULTS: No recurrent case was observed at three months after the operation. QOL of the patients was much improved based on the results of P-QOL, SF-36 and POP-Q. The results of questionnaire of patients' satisfaction were well correlated with other questionnaires. CONCLUSION: P-QOL can be expected as the linguistic validated questionnaire in Japanese version. Further studies will be needed for making linguistic validated P-QOL in Japanese version. PMID- 18404883 TI - [Comparison of clinical results between TUR-P and holmium laser enucleation of the prostate (HoLEP) based on the initial experience]. AB - OBJECTIVE: We compared the surgical results between holmium laser enucleation of the prostate (HoLEP) and transurethral resection of the prostate (TUR-P) for the treatment of men with benign prostatic hyperplasia (BPH). METHODS: A total of 87 patients with symptomatic BPH were analysed. HoLEP was performed on 46 men (mean age 68.2 +/- 7.5 years old) from December 2005 to February 2007, and TUR-P was performed on 41 men (mean age 69.2 +/- 7.3 years old) from April 2004 to March 2006. RESULTS: Both groups were comparable in terms of age, pre-operative IPSS, QOL index, urodynamic study results and prostate volume. During operation, decrease in hemoglobin was less in the HoLEP group than in the TUR-P group (1.15 +/- 1.2 vs 1.91 +/- 1.3 g/dl p < 0.05). The operation time was significantly longer in the HoLEP group than in the TUR-P group (161.9 +/- 65.0 vs. 118.3 +/- 36.9 minutes p < 0.001). Mean resected weight was 29.3 +/- 13.3 g (10-55) in the TUR-P group and 34.8 +/- 33.4 g (5-148) in the HoLEP group (p = 0.337). The catheterization period (52.1 +/- 29.6 vs. 115.2 +/- 27.5 hour p < 0.001) and hospital stay (6.6 +/- 2.3 vs. 9.4 +/- 2.2 days p < 0.001) were significantly shorter in the HoLEP group than in the TUR-P group. At follow up, Qmax, average flow rate and post void residual urine (PVR) in two groups improved significantly, and these parameters were not significantly different between the groups after 3 months. CONCLUSIONS: Both TUR-P and HoLEP were effective in relieving BOO. The estimated blood loss, a catheterization time and hospitalization were less or shorter in the HoLEP group. HoLEP may be a good alternative to the conventional transurethral electrocautery resection of the prostate for symptomatic BPH. PMID- 18404884 TI - [Case of cytomegalovirus colitis during standard chemotherapy for testicular cancer]. AB - CMV infection is uncommon in our practice except kidney transplantation. We report a case of cytomegalovirus enterocolitis during standard chemotherapy for testicular cancer. A 31-year-old male presented to his local urologist in January, 2006 with left scrotal swelling. He was diagnosed as left testicular tumor and underwent radical orchiectomy. Pathological examination showed Mixed germ cell tumor (seminoma, yolk sac, embryonal). Four months later, follow-up CT scan revealed multiple metastasis in lung and para aortic lymph nodes. The patient was referred to our institution for further consultation. During 3rd cycle of standard BEP chemotherapy, laboratory data showed severe bone marrow suppression and he subsequently presented prolonged spike fever with watery diarrhea. Despite intensive diagnostic examination including colon fiberscopy, those symptoms remained undiagnosed. One month later, the patient recovered with conservative treatment and serological data showed slightly elevated anti-CMV Ig M, Ig-G antibody. Immunohistological examination demonstrated CMV positive inclusion bodies in colorectal mucosa. Accordingly the patient's complication was finally diagnosed as CMV enterocolitis. Significant immunosuppression leading to severe colitis by CMV infection or reactivation can occur after standard chemotherapy. It is necessary to screen for CMV infection in patients with prolonged fever with diarrhea during chemotherapy. PMID- 18404885 TI - [Myxofibrosarcoma of the spermatic cord: a case report]. AB - A 77-year-old man visited our department with a painless mass in the left scrotum. He underwent left high orchiectomy with clinical diagnosis of testicular tumor. The tumor penetrated deeply into the internal inguinal ring. The tumor was resected as proximally as possible with positive surgical margin at the internal ring. Macroscopically, the left testis was intact in the resected specimen. Microscopic examination revealed that the tumor was consisted of myxoid stroma with spindle cells, and there was a large variation in histological findings. Immunohistchemical staining was positive with vimentin, and negative with alphaSMA, S-100, Myo D1, EMA, keratin, HMB45, and LCA. Accordingly, the tumor was diagnosed as myxofibrosarcoma, high-grade, of the spermatic cord. Postoperative CT scanning revealed multiple retroperitoneal lymph node metastases. He underwent radiation therapy, but soon developed lung metastases and pleural dissemination, and died 2 months after the operation. Myxofibrosarcoma, which had been called myxoid MFH is one of the most common soft tissue sarcomas. Myxofibrosarcoma or myxoid MFH of the spermatic cord is a rare disease. To our knowledge, only 7 cases have been reported in Japan to date. PMID- 18404886 TI - [Case of primitive neuroectodermal tumor of the kidney]. AB - A 28-year-old woman presented with right flank pain. A large, firm, fixed mass was palpable in the right side of the abdomen. Computed tomography revealed a solid mass of the right kidney with extension into the renal vein and inferior vena cava. The patient underwent right radical nephrectomy with en bloc resection of the inferior vena cava containing tumor thrombus and right adrenalectomy. Histologically the tumor consisted of small tumor cells with rosette formation. Immunohistochemical staining was positive for CD99 and NSE. Analysis with polymerase chain reaction (PCR) demonstrated the EWS/FLI1 fusion products resulting from a chromosomal translocation. These findings were consistent with primary renal primitive neuroectodermal tumor (PNET). Two months after surgery, multiple lung, liver and lymph node metastases were found. The patient received 2 cycles of chemotherapy with cisplatin, ifosfamide, etoposide, resulting in a partial remission. She subsequently received 1 cycle chemotherapy with paclitaxel and carboplatin, resulting in no response. The metastatic lung and liver diseases progressed and she died 5 months after diagnosis. PMID- 18404887 TI - [Factors associated with quality of life among patients with psoriasis. Comparison between psoriasis-specific QOL measures and generic QOL measures]. AB - OBJECTIVE: This study was conducted to determine factors associated with QOL in patients with psoriasis. For comparison, we used psoriasis-specific and generic QOL measures. METHODS: The targeted participants were 228 patients diagnosed as having psoriasis. Our questionnaire had two versions, the first one answered by the physician, and the second one answered by the patients themselves. The physician's questionnaire collected data concerning demographic characteristics, complications, health habits, affected body surface area; BSA, PASI and so on. The patients' questionnaire asked for Self-BSA, Self-PASI, PDI, SF-36 and so on. RESULTS: Questionnaires were returned by 216 patients. In consideration of some missing data on PDI and SF-36, the data for only 200 patients were used for this analysis. Concerning PDI, significant sex differences in mean scores were shown in areas of 'daily activities' and 'work'. Concerning SF-36, significant sex differences were shown in all areas except for 'general health'. Multiple regression analysis suggested that for male patients the PDI total score was related to age, PASI and Self-PASI; the SF-36 total score was related to PASI and Self-PASI. For female patients, although it was suggested that the PDI total score was related to PASI and Self-PASI, the SF-36 total score was related to Self-PASI only. On the whole, PDI was related much stronger for PASI and Self PASI than was SF-36. Furthermore, in female patients the Self-PASI score was strongly related to QOL scores. Therefore considering QOL for female patients, Self-PASI is possibly more useful than PASI. CONCLUSIONS: These findings indicate that severity of disease, sex and age are associated with QOL. PDI and Self-PASI are useful tools for the assessment and care of psoriasis patients considering QOL. PMID- 18404888 TI - [Present status of community-based HCV screening in Osaka City and evaluation of the utility of follow-up programs on hepatitis]. AB - OBJECTIVE: The present status of community-based HCV screening in Osaka City and the utility of follow-up programs on hepatitis were examined. METHOD: During a period of three years, from 2003-2005, 83, 458 persons who underwent HCV testing in a community-based HCV screening program in Osaka City were targeted. These persons were 40 years of age or older who opted to receive testing for HCV antibodies. To assist patients testing positive for HCV, hepatitis follow up programs were carried out. For cases where consent was given, the performance or nonperformance of a thorough examination could be ascertained with regard to whether or not a thorough examination results notification was sent from the medical institution. For patients who did not receive a notification, a health nurse checked whether a thorough examination was received, by telephone or visit, usually two times (after 3 months and after 6 months) on average. If an examination had not been received, the nurse suggested having one. Furthermore, the contents of the thorough examination and other details like procedure plans for all patients were obtained through the thorough examination results notification. From continuous diagnostic reports from the medical institutions, an assessment was given and the information was provided to each medical institution. For cases where consent was not given for the follow up programs, only the results of the thorough examination were available. RESULTS: The percentage of patients testing positive for HCV antibodies showed a yearly decline, with 3.9% in 2003, 3.8% in 2004, and 3.0% in 2005. The percentages of patients consenting to hepatitis follow up programs were 52.2% in 2003, 56.2% in 2004, and 59.1% in 2005. When comparing the percentage of patients receiving a thorough examination between those consenting to these programs and those not consenting, the consenting group was 82.6% and the non-consenting group was 37.5% in 2003, 77.1% and 37.7% in 2004, 78.0% and 34.3% in 2005, respectively. Consequently, the percentage of patients consenting to the examination significantly increased each year (P<0.001). From the diagnostic information which was sent, medical data could be provided for a total of 153 cases in 2003, 105 in 2004 and 58 in 2005. Furthermore, from among the contents, non-performance of imaging examination was most common with 107 cases (33.9%), followed by non identification of the HCV subtype, non-performance of a fixed quantity examination, and non-performance of an HCV-RNA qualitative examination to verify the presence of final stage virus. There were 50 cases where the follow-up was discontinued due to normal liver functions even though the subjects tested positive for HCV (15.8%). There were 15 cases where the follow-up was discontinued even though an HCV-RNA qualitative examination had not been performed (4.7%). Information that a followup was necessary was provided in those cases. CONCLUSION: In order for patients testing positive for HCV to receive an appropriate thorough examination and follow-up, assistance by verifying that the examination had been received and suggesting its necessity where this was not the case is useful. A more extensive performance of information provision to medical institutions is to be recommended. PMID- 18404889 TI - [Clinical assessment of community health nurses and the national government screening test for preventive health care among frail elderly living alone]. PMID- 18404890 TI - [Evaluation of public health center Websites from the viewpoints of content, usability, and accessibility]. PMID- 18404891 TI - [Degree of students' interest in community health activities before and after community nursing practice and associated factors]. PMID- 18404893 TI - [Advances in non-invasive measurement of brain function and clinical neurophysiology]. PMID- 18404892 TI - [Progress of radiotherapy and stereotactic radiotherapy to the early lung cancer]. PMID- 18404894 TI - Cell phone e-mail as a means to collect information on pregnancy and delivery: a pilot study in Japan. AB - BACKGROUND: The diffusion of cell phones is 91.9% in Japan. Especially, cell phone e-mail is now widely used as a media source in Japan. The authors of this research have been engaged in collecting basic data to establish a system for providing improved health information on pregnancy and delivery through cell phones for pregnant women and their family members. OBJECTIVE: In this study, we classified the trends of health information of pregnancy and delivery in which people desire to obtain through cell phone based on the characteristics of the information and, of each classification we revealed the generic characteristic (age, gender, gestational weeks and any other children), any social support, the types of cell phones or/and computers and their related use. METHODS: Anonymous questionnaires were distributed to the participants of delivery preparation workshops held at two places in the northern parts of Kyushu Area to collect data. The cell phone e-mail usage trends were measured by four-rated scales. To classify the characteristics of the information, a Principal Factor Analysis was conducted. To find out correlation between the information classified and the generic characteristics, Mann-Whitney's U-test was conducted. RESULTS: Out of two hundred and fifteen (215) respondents including 117 pregnant women and 98 husbands, 208 respondents possessed cell phones. Regarding the health information of pregnancy and delivery desired to obtain through cell phone e-mail, two factors, namely "medical information" and "non-medical information" have derived. Those who did not possess computers tended to use their cell phone e-mail to obtain both "medical information" (P = .002) and "non-medical information" (P = .02). While those who use cell phone e-mail almost everyday tended to use cell phone e-mail for obtaining "non-medical information" (P = .02). CONCLUSION: This study indicated that providing medical information and non-medical information through cell phone e-mail is an essential service, especially in Japan, where the diffusion of cell phones is far higher than that of computers. PMID- 18404895 TI - A pilot survey on working and health conditions in one Italian region: Venetia. Foreword. PMID- 18404896 TI - [Working and health conditions and preventive measures in a random sample of 5000 workers in the Veneto Region examined by telephone interview]. AB - BACKGROUND: A survey was carried out in 2000 by the European Foundation for the Improvement of Living and Working Conditions in a random sample of workers from 15 countries of the European Union in order to obtain information on occupational exposure, health problems and preventive measures taken at the workplaces. OBJECTIVES: To obtain similar information in workers in the Veneto Region and compare the results with those of the third European Survey on Working Conditions (ESWC). The results of the survey on Veneto Region workers were further analyzed, investigating the distribution by risk factors in each work sector, and the association between risk factors and reported health problems. METHODS: The ESWC questionnaire was adapted to the requirements of a telephone interview and a sample of 5000 workers (size based on the budget) between 15 and 64 years of age was randomly extracted from the regional list of telephone subscribers. The questionnaire was administered by trained interviewers. No statistical tests were used in the comparison between ESWC and Veneto Region results due to the lack of a priori hypotheses. Odds ratio (OR) was calculated in estimating the association between risk factors and symptoms; and p-value for OR trend across the increasing level of exposure was also obtained. RESULTS: Workers reported to be exposed for more than a quarter of their work time to: vibrations (20%), noise (19%), dusts, fumes vapours, chemicals (18%), repetitive hand/arm movements (50%), tiring/painful positions (46%); working at very high speed or tight deadlines (60%). 54.4% of the subjects reported working with computers. Taking as a reference the third ESWC in 2000, among Veneto Region workers in 2005 exposure was lower as regards physical, chemical and ergonomic risk factors, and similar as regards working at very high speed. The more frequently reported work-related health problems were: stress (26.9%), backache (17.8%), overall fatigue (11.9%), muscular pains in upper limbs (6.8%), headache (6.1%), sight problems (5.4%), anxiety (5.5%), muscular pain in lower limbs (4.3%), irritability (4.0%), hearing problems (2.3%). Except for stress, all symptoms/health problems were two-three times more frequently reported in the ESWC than in the Veneto Region survey where, conversely, the number of persons with at least one new sick- leave spell was higher. Lastly, there was no difference as regards preventive measures taken at the workplace: information on risks (78.2%), wearing personal protective equipment (28.7%), training paid by employer (28.7%). Among the Veneto Region workers, the most often reported risk factors were exposure to physical and chemical risk factors in industry/agriculture, and shift-work and working at very high speed in the services. The most commonly adopted preventive measures were information on risks and wearing of personal protective equipment in industry, and training in services. Moreover, among the Veneto Region workers, a significant exposure-dependent increase was reported for respiratory problems, allergies, dermatitis, hearing loss, accidents, back pain, pain in the upper and lower limbs, and headache. The risk of stress, anxiety, sleeping problems, stomach pain and headache increased when skills were not adequate to cope with job demand. In contrast, the perception of improved health conditions increased with increasing skill discretion, decision authority, social support (which are dimensions of control of job demand), but not with information on risk, training, or use of personal protective equipment. CONCLUSIONS: Data from the present survey provide useful insights on working and health conditions of workers in the Veneto Region, revealing problems that were subsequently investigated using other sources of information, as reported in the studies published elsewhere in this volume. PMID- 18404897 TI - [Occupational injuries and sickness absence: association with job demand, decision latitude, and life style in 2174 workers in the Veneto Region]. AB - BACKGROUND: Stress was the most frequent (26,9%) health problem reported in a survey on the perception of working and health conditions in 5000 workers in the Veneto Region. OBJECTIVES: The aim of the study was to investigate in the Veneto Region the association between occupational stress and events occurred in the previous 12 months: occupational accidents, or sickness absence for 10 or more consecutive days. METHODS: Perceived occupational stress is correlated, according to Karasek's model, to high job demand (JD) and low decision latitude (DL). Using Karasek's questionnaire (to which questions on smoking and alcohol consumption were added), we examined 2174 subjects working in 30 companies with between 10 and 500 employees, who belonged to the occupational categories of industry and services that are more prevalent in the Veneto Region. The questionnaire was administered by the occupational physician during health surveillance. The subjects were classified on the basis of current exposure to psychosocial factors or, for subjects reporting an event, their exposure at that time. We identified the tertiles of JD and DL; data were submitted to the analysis of multiple logistic regression, estimating odds ratio (OR) and 95% confidence interval (CI). The population attributable risk (PAR) was calculated using the formula (pc (OR 1)/OR), where pc is the fraction of exposed cases. RESULTS: An elevated risk of occupational accidents was found in subjects with regular consumption of alcohol (OR = 2.0; IC = 1.2-3.5), in smokers smoking 10-20 (2.3; 1.3-3.8) or >20 cigarettes/day (3.8; 1.8-7.9), in the highest tertile of JD (2.29; 1.35-3.89) and in the lowest tertile of DL (1.6; 1.0-2.6). PAR was 37.6% for occupational factors (high JD and low DL), 44.5%for non-occupational factors (cigarette smoking and alcohol consumption), and 82.1% overall. The risk of sickness absence increased in subjects smoking 10-20 cigarettes (1.63 = 1.1-2.40), in the highest tertile of JD (1.5; 1.0-2.2) and in the lowest tertile of DL (1.6; 1.1-2.2). PAR was 26.1% for occupational factors (high JD and low DL), 7.6% for non occupational factors (smoking), and 30.4% overall. While the risk of sick absence increased mainly with the reduction of DL, the risk of occupational accidents increased with increasing JD and, to a lesser extent, with decreasing DL. The current approach to accident prevention is based only on technical and administrative aspects, in spite of the fact that 80% of accidents are not attributable to malfunctioning of machinery. Injury prevention should address technical, personal and psychosocial risk factors together as a whole. PMID- 18404898 TI - [Comparison of four measurements of sickness absence in workers in the Veneto Region: interpretation and applicability. Data from Italian National Social Security Institute, 1997-2002]. AB - BACKGROUND: In 2005, 52% of workers in the Veneto Region reported one or more sick leave spells in the previous 12 months, compared with 16% reported in the European Survey on Working Condition in 2000 (ESWC), although health conditions were better in the Veneto Region than in the ESWC. OBJECTIVES: Since the above findings were based on answers to a questionnaire, we used an objective source of information in order to further investigate the problem. METHODS: The source of data consisted of the sick leave records of workers in the Veneto Region, 1997 2002, which were obtained from the Italian National Social Security Institute (INPS). Four measurements were used: (1) frequency = number of sick-leave spells during the study period/number of insured persons at risk for sickness absence during the study period; (2) cumulative incidence = number of persons with at least one new sick-leave spell during the study period/number of insured persons at risk for sickness absence during the study period; (3) sick-leave rate = number of sick-leave days during the study period/number of insured persons at risk for sickness absence during the study period; (4) duration of absence = number of sick-leave days in new sick leave spells during the study period/number of new sick-leave spells during the study period. Cumulative incidence could only be analyzed by statistical methods (binomial regression). RESULTS: The first three measurements gave overlapping results. The values were highest in industry, lowest in agriculture, and intermediate in crafts and services; they were highest in middle-aged subjects and lowest in younger and older individuals; the time trend increased up to 1999 and then decreased slowly. Cumulative incidence was 52.0%, 38.0%, 36.4% and 22.9%, in industry, services, crafts, and agriculture, respectively, and 42.8% in the total population. The results of binomial regression confirmed the above pattern, as well as an excess in women, in white collar workers, and in various occupational categories which, however, were probably not due to occupational exposures since in each occupational category the risk was always higher in industry than in crafts. By contrast, the sick leave rate was higher in crafts and agriculture and lower in services and industry; it increased exponentially with age and did not show real differences between males and females. CONCLUSIONS: The choice of measurements influences the results; duration of absence could be used to detect areas of suspected work related diseases, while cumulative incidence might be more helpful to detect areas of suspected absenteeism/presenteeism. PMID- 18404899 TI - [Association between causes of lNPS compensated disability and occupational category (Veneto Region, 1994-2002): a surveillance method for occupational diseases?]. AB - BACKGROUND: According to Italian law, workers are insured against disability through the National Social Security Institute (INPS), whose records contain information on diseases causing disability and occupational histories. OBJECTIVES: Using the INPS data, our objective was to identify any excess risk of disabling diseases in relation to the occupational categories. METHODS: Insurance covers all private sector employees, a small proportion of public sector employees, agricultural labourers, domestic workers, self-employed workers (craftsmen, commercial dealers,farmers), and few other occupational categories. For each insured worker, a database was created containing registry data, occupational history and, for compensated workers, the disease code, for each year from 1994 to 2002. A cohort study design was adopted, in which references were self-employed workers (with mixed exposure) to whom different categories of employees in agriculture, industry, crafts, and services were compared. Data were analyzed by means of Poisson regression, estimating the Incidence Rate Ratio (IRR) and confidence interval (CI) at 99.99%, instead of 95%, in order to set the threshold of error for the entire study at 0.05. RESULTS: In the construction industry there were significantly high risks of disability in both industry and crafts for tumours (industry: IRR = 2.07; IC = 1. 67-2.57; crafts: 2.57; 1.89 3.18), circulatory disorders (industry: IRR = 2.24; IC = 1.65-3.04; crafts.: 3.06; 2.16-4.32), and bone and joint diseases (industry. IRR = 5.0; IC = 3.15 7.94; crafts: 6.58; 5.04-8.59). CONCLUSIONS: The advantage of this approach is to recruit a large number of subjects at limited cost. The procedure here proposed is a mainly exploratory approach aimed at establishing new study hypotheses: disability, in fact, is acknowledged by INPS when its cause is not occupational according to the current literature. PMID- 18404900 TI - [Safety and health in workers employed in industry. Data from Industrial Accidents Compensation Board (INAIL) and National Social Security Institute (INPS), Veneto Region, 1994-2002]. AB - BACKGROUND: A decreasing time trend for occupational injuries and sickness absence would be the effect of the new legislation (D.Lgs. 626/94 and successive laws) on prevention in occupational settings. Conversely, the reduction of INPS disability would reflect a health improvement due to non-occupational causes. OBJECTIVES: The aim of the study was to investigate the efficacy of the new legislation among employees in industry (where the law was mainly applied), via the time trend of three standardized rates in the Veneto Region. METHODS: The numerator for the rate of occupational accidents (cases occurring in industry workers in the Veneto Region, broken down for sex, age and calendar years) was supplied by INAIL. The denominator for the above rate, as well as numerators and denominators for disability and sickness absence were supplied by INPS. Data were available from 1994 to 2002 for accidents and disability, and from 1997 to 2002 for sickness absence. In every year from 1994 to 2002, the rates were standardized for age and sex with the direct method, using an internal "standard" population. The time trend of year-specific standardized rates was analyzed by Joinpoint regression software. RESULTS: Among industrial workers in the Veneto Region, occupational accidents increased by 0.4% yearly, while disability decreased by 2.56% from 1994 to 2002. Sick absence increased up to 1999, then decreased. CONCLUSIONS: This epidemiological pattern is difficult to explain. The increase in accidents could be due to the increase of non-European Union workers and/or to the fact that accidents on the way to or from work were recognized as occupational accidents by INAIL starting from 2000. Both these phenomena could have contributed to increase the rate that was otherwise diminishing. On the other hand, this same situation could be due to insufficient efficacy of the legislation (D.Lgs. 626/94 and successive laws) for preventing occupational accidents and diseases. PMID- 18404901 TI - [A procedure for estimating the rate of occupational accidents in non-European Union workers with irregular immigrant status]. AB - BACKGROUND: Statistics on occupational accidents provided by the Italian Institute for Occupational Diseases and Accidents (INAIL, Italian acronym) include only events that occurred in workers with regular employment status. OBJECTIVES: The aim of the study was to establish a procedure in order to estimate the rate of occupational accidents in non-European-Union (non-EU) workers with irregular employment status and/or irregular immigrant status. METHODS: The sources of data were the clinical records of the Emergency Department of San Bonifacio Hospital, and the population data of District 4 of Local Health Authority 20 of Verona, which was considered the catchment area of this hospital. RESULTS: Among 419 cases of accidents occurred in the numerator of the rate. The denominator of the rate was estimated by calculating: (1) the subjects of working age resident in District 4 (= 83714); (2) the total number of non-EU workers, assuming that the percentage was similar to that in San Bonifacio Municipality (= 0.115); the number of irregular non-EU workers, assuming that the percentage was similar to that in north-eastern Italy (= 0.103). Non-EU workers with irregular employment status and/or irregular immigrant status should, according to these calculations, be 992 (= 83714 x 0.115 x 0.103). The rate- 147.2 (= 146/992) occupational accidents per 1000 irregular non-EU workers--is more than twice as high as that calculated in 2004 in Italy in regular non-EU workers (approximately 65 accidents per 1000). The difference can be explained by the fact that irregular workers find employment mainly in agriculture, building and the metallurgic industry, which have a high frequency of accidents, and are more willing to accept risky work and longer work shifts. CONCLUSIONS: On the assumption that the rate of occupational accidents in the 500,000 irregular workers living in Italy in 2004 was 147.2 per 1000 (as in the catchment area of the San Bonifacio Hospital), the number of accidents would be 73,600, against the 116,000 that occurred among regular non-EU workers in 2004 according to INAIL. Official INAIL statistics on occupational accidents therefore show a considerable underestimation. PMID- 18404902 TI - Disruptive physicians: sound more familiar than you thought? PMID- 18404903 TI - Auricular injury with deformity following delivery by cesarean section. PMID- 18404904 TI - Sudden onset of sensorineural hearing loss secondary to intralabyrinthine hemorrhage: MRI findings. PMID- 18404905 TI - Gout. PMID- 18404906 TI - Granular cell tumor of the esophagus presenting as a duplication cyst. PMID- 18404907 TI - The "pancake flip": management of extreme internal curvature of the lower lateral cartilage. PMID- 18404908 TI - Effect of cochlear implantation on nasality in children. AB - Hypernasality is a commonly perceived characteristic of speech in deaf adults and children, but the mechanism of this abnormal nasal resonance is poorly understood. The impact of cochlear implantation on nasalance measures in children with severe auditory deprivation has not been previously reported. We conducted a study of nasality in 6 deaf children who had undergone cochlear implantation. Voice recordings were obtained before surgery and 6 months after activation of the implants. The MacKay-Kummer SNAP Test--which consists of a syllable repetition subtest and a picture-cued subtest--was used to obtain nasalance scores for oral (bilabial, alveolar, velar, and sibilant) and nasal phonemes. Before cochlear implantation, mean nasalance scores were significantly higher than normal during the production of oral phonemes for both subtests (p < or = 0.05). Six months after activation, the nasalance measures for all components of the syllable-repetition subtest had been restored to within 1 standard deviation of normal. For all oral phonemes of the picture-cued subtest, the elevated nasalance scores were consistently lower after cochlear implant activation, although the difference was statistically significant only for velar tasks. Nasalance scores for nasal phonemes were within 1 standard deviation of normal both before and after implant activation. Our study showed that cochlear implantation partially corrects elevated nasalance measures. Disturbances in nasal resonance may be caused in part by the inability of deaf speakers to monitor velopharyngeal valving with auditory feedback. The trend toward improved nasalance scores after implantation highlights the role of auditory feedback in monitoring velopharyngeal function. Visual biofeedback may be required to further normalize hypernasal speech in profoundly deaf children. PMID- 18404909 TI - Hearing loss and cerebrospinal fluid pressure: case report and review of the literature. AB - A decrease in cerebrospinal fluid pressure may result in an endolymphatic hydrops through a patent cochlear aqueduct or through the fundus of the internal auditory canal. This hydrops typically leads to low-frequency sensorineural hearing loss. We describe the case of a man who presented with a subjective and objective hearing loss in addition to a headache 4 days after he had undergone a dural puncture. We treated him with a standard epidural blood patch. Immediately after treatment, his hearing improved and his headache resolved. PMID- 18404910 TI - Auricular ossification resulting in external auditory canal stenosis. AB - We report the case of a 49-year-old man who presented with left aural fullness, hearing loss, and a stenotic left external auditory meatus. Physical examination and computed tomography revealed the presence of an idiopathic ossification of the cartilaginous auricle and external auditory canal. The patient was successfully treated with meatoplasty. Pathologic examination of the removed specimen revealed osseous metaplasia. PMID- 18404911 TI - Rhinolithiasis with a nasal polyp: a case report. AB - Rhinoliths are uncommon mineralized masses that form as a result of calcification of an endogenous or exogenous nidus. The most common manifestations of rhinolithiasis are unilateral nasal discharge, nasal obstruction, and facial pain. The diagnosis is made by nasal endoscopy and computed tomography. The differential diagnosis includes chronic inflammation, osteomyelitis, benign tumors (e.g., calcified nasal polyps, ossifying fibromas, osteomas, and chondromas), and malignant tumors (e.g., osteosarcomas, chondrosarcomas, and squamous cell carcinomas). Rhinoliths may cause rhinosinusitis, erosion of the nasal septum and medial wall of the maxillary sinus, and perforations of the palate. To the best of our knowledge, the occurrence of a nasal polyp associated with rhinolithiasis has not been previously reported in the English-language literature. In this article, we describe such a case. PMID- 18404912 TI - An atypical case of fatal zygomycosis: simultaneous cutaneous and laryngeal infection in a patient with a non-neutropenic solid prostatic tumor. AB - We describe what we believe is the first reported case of simultaneous highly invasive cutaneous and laryngopharyngeal zygomycosis in a non-neutropenic, nondiabetic but immunosuppressed patient with prostate cancer. An invasive fungal process was not suspected until late in the patient's hospital course; when it was, a tracheotomy and direct laryngoscopic biopsies were performed. Unresectable invasive zygomycosis with Rhizopus rhizopodiformis was diagnosed. The patient was managed with liposomal amphotericin B initially and later with palliative medical therapy until he died. This case emphasizes the need for a rapid and specific diagnosis with timely introduction of appropriate antifungal management, particularly now that voriconazole is frequently used as empiric prophylaxis against aspergillosis in high-risk patients. PMID- 18404913 TI - Retropharyngeal hematoma as a complication of anticoagulation therapy. AB - Reports in the literature of retropharyngeal hematoma as a complication of anticoagulation therapy are rare. When this complication does occur, it can become life-threatening if the airway is compromised. However, no consensus exists as to which approach--intubation, tracheotomy, or conservative therapy--is best for managing the airway in these cases. We report a case of retropharyngeal hematoma that occurred as a sequela to a trivial blunt trauma in a 48-year-old man who had been undergoing anticoagulation therapy with warfarin. The hematoma had caused airway obstruction, and the patient was hospitalized. He was treated conservatively, and the hematoma slowly resolved over the course of 2 weeks. On the basis of our experience and the findings of our literature review, we suggest that conservative management can be initiated for small nonexpanding hematomas that do not seriously compromise the airway. Securing the airway with intubation or tracheotomy should be reserved for patients who are in serious respiratory distress; the choice between intubation and tracheotomy should be made on an individual basis. PMID- 18404914 TI - Dentigerous cyst of the maxilla with impacted tooth displaced into orbital rim and floor. AB - We report a case of dentigerous cyst of the maxilla and maxillary sinus that caused the ectopic displacement of an unerupted tooth into the orbital rim and floor. After an incisional biopsy, marsupialization of the lesion promoted its involution and stimulated osteogenesis. This in turn simplified the surgical enucleation of the specimen and removal of the unerupted tooth without the excessive loss of the bony contours of the maxilla. PMID- 18404916 TI - Congenital midline cervical cleft: a report of two cases. AB - Congenital midline cervical cleft is an uncommon malformation of the anterior neck. We report the cases of two patients with such an anomaly who had been born within 10 days of each other at a regional hospital in upstate New York. Fewer than 100 cases of congenital midline cervical cleft have been reported in the literature, and to the best of our knowledge, none of the other reported cases occurred so close to each other in terms of time and location. Diagnoses in both cases were established by clinical observation soon after birth. No likely epidemiologic commonalities between the two patients were found. Head and neck surgeons should be familiar with these malformations because early surgical excision is critical. Both of our patients were surgically treated at an early age, one with a Z-plasty and one with a straight closure. In both cases, the patients exhibited good functional and cosmetic results at 5 years of age. PMID- 18404915 TI - Lingual hematoma and heparin-induced thrombocytopenia: a case report. AB - Lingual hematoma is a rare but potentially fatal cause of upper airway obstruction. Patients receiving anticoagulants such as heparin can suffer from significant complications of these medications. Not only does heparin exert effects directly on the coagulation cascade, but it has the potential to cause thrombocytopenia by stimulating formation of antibodies against platelets. We present the case of a patient being treated with heparin for a deep-vein thrombosis, who subsequently developed heparin-induced thrombocytopenia and lingual hematoma, necessitating tracheotomy. PMID- 18404917 TI - Formation of an airway foreign body during removal of a metal tracheostomy tube: a case report. AB - Tracheostomy tube changes are often performed in the clinic or at the bedside without complication. We report a case in which the removal of a long-term metal tube in a clinic setting was complicated by the presence of fibrinous debris on the tube. The debris had accumulated over a period of at least 3 years. The patient was taken to the operating room, and the tube was removed. However, the fibrinous debris remained behind, leaving an airway foreign body. The foreign body was subsequently removed with an optical foreign-body extraction forceps under direct visualization. Based on our experience and on the limited amount of information available in the literature, we recommend that long-term metal tracheostomy tubes be changed not in the clinic but in the operating room, where potential complications can be more easily managed. PMID- 18404918 TI - Point/Counterpoint. Miniature x-ray tubes will ultimately displace Ir-192 as the radiation sources of choice for high dose rate brachytherapy. PMID- 18404919 TI - An automated intensity-weighted brachytherapy seed localization algorithm. AB - Brachytherapy has proven to be an effective treatment for various forms of cancer, whereby radioactive material is inserted directly into the body to maximize dosage to malignant tumors while preserving healthy tissue. In order to validate the preoperative or intraoperative dosimetric model, a postimplant evaluation procedure is needed to ensure that the locations of the implanted seeds are consistent with the planning stage. Moreover, development of an automated algorithm for seed detection and localization is necessary to expedite the postimplant evaluation process and reduce human error. Most previously reported algorithms have performed binary transforms on images before attempting to localize seeds. Furthermore, traditional approaches based upon three dimensional seed shape parameterization and matching require high resolution imaging. The authors propose a new computationally efficient algorithm for automatic seed localization for full three-dimensional, low-resolution data sets that directly applies voxel intensity to the estimation of both seed centroid location and angular seed orientation. Computer simulations, phantom studies, and in vivo computed tomography prostate seed imaging results show that the proposed algorithm can produce reliable results even for low-resolution images. PMID- 18404920 TI - Anniversary paper: Role of medical physicists and the AAPM in improving geometric aspects of treatment accuracy and precision. AB - The last 50 years have seen great advances in the accuracy of external beam radiation therapy. Geometrical uncertainties have been reduced from a centimeter or more in presimulation, skin-mark guided days to 1-2 mm in today's image-guided radiation therapy treatments. Medical physicists, with the support and guidance of the American Association of Physicists in Medicine (AAPM), have been, and continue to be, at the forefront of research, development and clinical implementation in this area. This article reviews some of the major contributions of physicists to the improvement of treatment accuracy and precision, and speculates as to what the future may bring. PMID- 18404921 TI - Seed-based transrectal ultrasound-fluoroscopy registration method for intraoperative dosimetry analysis of prostate brachytherapy. AB - Prostate brachytherapy is an effective treatment option for early-stage prostate cancer. During a prostate brachytherapy procedure, transrectal ultrasound (TRUS) and fluoroscopy imaging modalities complement each other by providing good visualization of soft tissue and implanted seeds, respectively. Therefore, the registration of these two imaging modalities, which are readily available in the operating room, could facilitate intraoperative dosimetry, thus enabling physicians to implant additional seeds into the underdosed portions of the prostate while the patient is still on the operating table. It is desirable to register TRUS and fluoroscopy images by using the seeds as fiducial markers. Although the locations of all the implanted seeds can be reconstructed from three fluoroscopy images, only a fraction of these seeds can be located in TRUS images. It is challenging to register the TRUS and fluoroscopy images by using the identified seeds, since the correspondence between them is unknown. Furthermore, misdetection of nonseed structures as seeds can lead to the inclusion of spurious points in the data set. We developed a new method called iterative optimal assignment (IOA) to overcome these challenges in TRUS-fluoroscopy registration. By using the Hungarian method in an optimization framework, IOA computes a set of transformation parameters that yield the one-to-one correspondence with minimum cost. We have evaluated our registration method at varying noise levels, seed detection rates, and number of spurious points using data collected from 25 patients. We have found that IOA can perform registration with an average root mean square error of about 0.2 cm even when the seed detection rate is only 10%. We believe that IOA can offer a robust solution to seed-based TRUS-fluoroscopy registration, thus making intraoperative dosimetry possible. PMID- 18404922 TI - Calibration of megavoltage cone-beam CT for radiotherapy dose calculations: correction of cupping artifacts and conversion of CT numbers to electron density. AB - Megavoltage cone-beam CT (MV CBCT) is used for three-dimensional imaging of the patient anatomy on the treatment table prior to or just after radiotherapy treatment. To use MV CBCT images for radiotherapy dose calculation purposes, reliable electron density (ED) distributions are needed. Patient scatter, beam hardening and softening effects result in cupping artifacts in MV CBCT images and distort the CT number to ED conversion. A method based on transmission images is presented to correct for these effects without using prior knowledge of the object's geometry. The scatter distribution originating from the patient is calculated with pencil beam scatter kernels that are fitted based on transmission measurements. The radiological thickness is extracted from the scatter subtracted transmission images and is then converted to the primary transmission used in the cone-beam reconstruction. These corrections are performed in an iterative manner, without using prior knowledge regarding the geometry and composition of the object. The method was tested using various homogeneous and inhomogeneous phantoms with varying shapes and compositions, including a phantom with different electron density inserts, phantoms with large density variations, and an anthropomorphic head phantom. For all phantoms, the cupping artifact was substantially removed from the images and a linear relation between the CT number and electron density was found. After correction the deviations in reconstructed ED from the true values were reduced from up to 0.30 ED units to 0.03 for the majority of the phantoms; the residual difference is equal to the amount of noise in the images. The ED distributions were evaluated in terms of absolute dose calculation accuracy for homogeneous cylinders of different size; errors decreased from 7% to below 1% in the center of the objects for the uncorrected and corrected images, respectively, and maximum differences were reduced from 17% to 2%, respectively. The presented method corrects the MV CBCT images for cupping artifacts and extracts reliable ED information of objects with varying geometries and composition, making these corrected MV CBCT images suitable for accurate dose calculation purposes. PMID- 18404923 TI - Model-independent, multimodality deformable image registration by local matching of anatomical features and minimization of elastic energy. AB - With respect to the demands of adaptive and 4D-radiotherapy applications, an algorithm is proposed for a fully automatic, multimodality deformable registration that follows the concept of translational relocation of regularly distributed image subvolumes governed by local anatomical features. Thereby, the problem of global deformable registration is broken down to multiple independent local registration steps which allows for straightforward parallelization of the algorithm. In a subsequent step, possible local misregistrations are corrected for by minimization of the elastic energy of the displacement field under consideration of image information. The final displacement field results from interpolation of the subvolume shift vectors. The algorithm can employ as a similarity measure both the correlation coefficient and mutual information. The latter allows the application to intermodality deformable registration problems. The typical calculation time on a modern multiprocessor PC is well below 1 min, which facilitates almost-interactive, "online" usage. CT-to-MRI and CT-to-cone beam-CT registrations of head-and-neck data sets are presented, as well as inhale to-exhale registrations of lung CT data sets. For quantitative evaluation of registration accuracy, a virtual thorax phantom was developed; additionally, a landmark-based evaluation on four lung respiratory-correlated CT data sets was performed. This consistently resulted in average registration residuals on the order of the voxel size or less (3D-residuals approximately 1-2 mm). Summarizing, the presented algorithm allows an accurate multimodality deformable registration with calculation times well below 1 min, and thus bears promise as a versatile basic tool in adaptive and 4D-radiotherapy applications. PMID- 18404924 TI - Geometric interpretation of the gamma dose distribution comparison technique: interpolation-free calculation. AB - The gamma dose comparison tool has been used by numerous investigators to quantitatively compare multidimensional dose distributions. The gamma tool requires the specification of dose and distance-to-agreement (DTA) criteria for acceptable variations between the dose distributions. The tool then provides a comparison that simultaneously evaluates the dose difference and distance to agreement of the two dose distributions. One of the weaknesses of the tool is that the comparison requires one of the dose distributions to have a relatively high spatial resolution, with points spaced significantly closer than the DTA criterion. The determination of gamma involves an exhaustive search process, so the computation time is significant if an accurate gamma is desired. The reason for the need for high spatial resolution lies with the fact that the gamma tool measures the closest point in one of the dose distributions (the evaluated distribution) with individual points of the other distribution (the reference distribution) when the two distributions are normalized by the dose difference and DTA criteria for the dose and spatial coordinates, respectively. The closest point in the evaluated distribution to a selected reference distribution point is the value of gamma at that reference point. If individual evaluated dose distribution points are compared, the closest point may not accurately reflect the closest value of the evaluated distribution as if it were interpolated on an infinite resolution grid. Therefore, a reinterpretation of the gamma distribution as the closest geometric distance between the two distributions is proposed. This is conducted by subdividing the evaluated distribution into simplexes; line segments, triangles, and tetrahedra for one, two, and three-dimensional (3D) dose distributions. The closest distance between any point and these simplexes can be straightforwardly computed using matrix multiplication and inversion without the need of interpolating the original evaluated distribution. While an exhaustive search is still required, not having to interpolate the evaluated distribution avoids the drastic growth of calculation time incurred by interpolation and makes the gamma tool more practical and more accurate. In our experiment, the geometric method accurately computes gamma distributions between 3D dose distributions on a 200 x 200 x 50 grid within two minutes. PMID- 18404925 TI - Computerized analysis of prostate lesions in the peripheral zone using dynamic contrast enhanced MRI. AB - A novel automated computerized scheme has been developed for determining a likelihood measure of malignancy for cancer suspicious regions in the prostate based on dynamic contrast-enhanced magnetic resonance imaging (MRI) (DCE-MRI) images. Our database consisted of 34 consecutive patients with histologically proven adenocarcinoma in the peripheral zone of the prostate. Both carcinoma and non-malignant tissue were annotated in consensus on MR images by a radiologist and a researcher using whole mount step-section histopathology as standard of reference. The annotations were used as regions of interest (ROIs). A feature set comprising pharmacokinetic parameters and a T1 estimate was extracted from the ROIs to train a support vector machine as classifier. The output of the classifier was used as a measure of likelihood of malignancy. Diagnostic performance of the scheme was evaluated using the area under the ROC curve. The diagnostic accuracy obtained for differentiating prostate cancer from non malignant disorders in the peripheral zone was 0.83 (0.75-0.92). This suggests that it is feasible to develop a computer aided diagnosis system capable of characterizing prostate cancer in the peripheral zone based on DCE-MRI. PMID- 18404926 TI - Correction of megavoltage cone-beam CT images for dose calculation in the head and neck region. AB - Megavoltage cone-beam computed tomography (MVCBCT) imaging systems are now available for image-guided radiation therapy delivery and verification. In order to use the three-dimensional anatomical information for dose calculation, the MVCBCT image must provide accurate electron density. This work proposes a new method that has been developed to correct for the cupping and missing data artifacts seen on MVCBCT images of the head and neck region. It uses a conventional kilovoltage CT (kVCT) image as a reference for electron density and rigid registration with a MVCBCT image to obtain correction factors. Dose calculations performed on MVCBCT images corrected with the proposed method agree with calculations done on kVCT images within +/- 1% on phantoms. With patients images the agreement is within +/- 13% above the shoulders and +/- 5% below the shoulder line. This level of dose calculation accuracy allows the use of MVCBCT images for dose verification purposes. PMID- 18404927 TI - A detailed study on the use of polynomial functions for modeling geometric distortion in magnetic resonance imaging. AB - The use of polynomial functions for modeling geometric distortion in magnetic resonance imaging (MRI) that arises from scanner's hardware imperfection is studied in detail. In this work, the geometric distortion data from four representative MRI systems were used. Modeling of these data using polynomial functions of the fourth, fifth, sixth, and seventh orders was carried out. In order to investigate how this modeling performed for different size and shape of the volume of interest, the modeling was carried out for three different volumes of interest (VOI): a cube, a cylinder, and a sphere. The modeling's goodness was assessed using both the maximum and mean absolute errors. The modeling results showed that (i) for the cube VOI there appears to be an optimal polynomial function that gives the least modeling errors and the sixth order polynomial was found to be the optimal polynomial function for the size of the cubic VOI considered in the present work; (ii) for the cylinder VOI, all four polynomials performed approximately equally well but a trend of a slight decrease in the mean absolute error with the increasing order of the polynomial was noted; and (iii) for the sphere VOI, the maximum absolute error showed some variations with the order of the polynomial, with the fourth order polynomial producing the smallest maximum absolute errors. It is further noted that extrapolation could lead to very large errors so any extrapolation needs to be avoided. A detailed analysis on the modeling errors is presented. PMID- 18404928 TI - Iterative sorting for four-dimensional CT images based on internal anatomy motion. AB - Current four-dimensional (4D) computed tomography (CT) imaging techniques using multislice CT scanners require retrospective sorting of the reconstructed two dimensional (2D) CT images. Most existing sorting methods depend on externally monitored breathing signals recorded by extra instruments. External signals may not always accurately capture the breathing status and may lead to severe discontinuity artifacts in the sorted CT volumes. This article describes a method to find the temporal correspondences for the free-breathing multislice CT images acquired at different table positions based on internal anatomy movement. The algorithm iteratively sorts the CT images using estimated internal motion indices. It starts from two imperfect reference volumes obtained from the unsorted CT images; then, in each iteration, thorax motion is estimated from the reference volumes and the free-breathing CT images. Based on the estimated motion, the breathing indices as well as the reference volumes are refined and fed into the next iteration. The algorithm terminates when two successive iterations attain the same sorted reference volumes. In three out of five patient studies, our method attained comparable image quality with that using external breathing signals. For the other two patient studies, where the external signals poorly reflected the internal motion, the proposed method significantly improved the sorted 4D CT volumes, albeit with greater computation time. PMID- 18404929 TI - Evaluation of beam wobbling methods for heavy-ion radiotherapy. AB - The National Institute of Radiological Sciences (NIRS) has extensively studied carbon-ion radiotherapy at the Heavy-Ion Medical Accelerator in Chiba (HIMAC) with some positive outcomes, and has established its efficacy. Therefore, efforts to distribute the therapy to the general public should be made, for which it is essential to enable direct application of clinical and technological experiences obtained at NIRS. For widespread use, it is very important to reduce the cost through facility downsizing with minimal acceleration energy to deliver the HIMAC equivalent clinical beams. For the beam delivery system, the requirement of miniaturization is translated to reduction in length while maintaining the clinically available field size and penetration range for range-modulated uniform broad beams of regular fields that are either circular or square for simplicity. In this paper, we evaluate the various wobbling methods including original improvements, especially for application to the compact facilities through the experimental and computational studies. The single-ring wobbling method used at HIMAC is the best one including a lot of experience at HIMAC but the residual range is a fatal problem in the case of a compact facility. On the other hand, uniform wobbling methods such as the spiral and zigzag wobbling methods are effective and suitable for a compact facility. Furthermore, these methods can be applied for treatment with passive range modulation including respiratory gated irradiation. In theory, the choice between the spiral and zigzag wobbling methods depends on the shape of the required irradiation field. However, we found that it is better to use the zigzag wobbling method with transformation of the wobbling pattern even when a circular uniform irradiation field is required, because it is difficult to maintain the stability of the wobbler magnet due to the rapid change of the wobbler current in the spiral wobbling method. The regulated wobbling method, which is our improvement, can well expand the uniform irradiation field and lead to reducing the power requirement of the wobbler magnets. Our evaluations showed that the regulated zigzag wobbling method is the most suitable method for use in currently designed compact carbon-therapy facilities. PMID- 18404930 TI - The x-ray light valve: a potentially low-cost, digital radiographic imaging system-concept and implementation considerations. AB - New x-ray radiographic systems based on large-area flat-panel technology have revolutionized our capability to produce digital x-ray images. However, these imagers are extraordinarily expensive compared to the systems they are replacing. Hence, there is a need for a low-cost digital imaging system for general applications in radiology. A novel potentially low-cost radiographic imaging system based on established technologies is proposed-the X-Ray Light Valve (XLV). This is a potentially high-quality digital x-ray detector made of a photoconducting layer and a liquid-crystal cell, physically coupled in a sandwich structure. Upon exposure to x rays, charge is collected on the surface of the photoconductor. This causes a change in the optical properties of the liquid crystal cell and a visible image is generated. Subsequently, it is digitized by a scanned optical imager. The image formation is based on controlled modulation of light from an external source. The operation and practical implementation of the XLV system are described. The potential performance of the complete system and issues related to sensitivity, spatial resolution, noise, and speed are discussed. The feasibility of clinical use of an XLV device based on amorphous selenium (a-Se) as the photoconductor and a reflective electrically controlled birefringence cell is analyzed. The results of our analysis indicate that the XLV can potentially be adapted to a wide variety of radiographic tasks. PMID- 18404931 TI - The effective point of measurement of ionization chambers and the build-up anomaly in MV x-ray beams. AB - A precision experimental investigation of the effective point of measurement (EPOM) of ion chambers in megavoltage beams has been carried out. A one dimensional scanning phantom system was developed with an overall accuracy in the positioning of a chamber of better than 0.15 mm. Depth-dose data were acquired for a 25 MV beam from an Elekta Precise linac (field sizes of 10 x 10 cm and 25 x 25 cm) for measurement depths in the range 0.6-6 cm. The results confirmed the Monte Carlo calculations of an earlier theoretical investigation by Kawrakow [Med. Phys. 33, 1829-1839 (2006)] that the standard shift for cylindrical chambers, recommended in dosimetry protocols of -0.6r (where r is the internal radius of the cavity), is incorrect. A wide range of ion chambers were investigated and it was found that errors of up to 1.4 mm could occur for certain chamber designs (although typical errors for common chambers were around 0.5 mm). A comparison between measurements and Monte Carlo simulations showed that once the correct EPOM is used, the details of the linac geometry are correct, and the parameters of the electron beam striking the bremsstrahlung target have been adequately determined, the EGSnrc Monte Carlo package is capable of reproducing the experimental data to 0.2 mm or better. The investigation also confirmed that for the highest accuracy depth-dose curves in megavoltage photon beams one should use a well-guarded parallel-plate ion chamber. Three chamber designs were tested here and found to be satisfactory-the Scanditronix-Wellhofer NACP-02, PTW Roos and Exradin All. PMID- 18404932 TI - The x-ray light valve: a potentially low-cost, digital radiographic imaging system--a liquid crystal cell design for chest radiography. AB - Digital x-ray radiographic systems are desirable as they offer high quality images which can be processed, transferred, and stored without secondary steps. However, current clinical systems are extraordinarily expensive in comparison to film-based systems. Thus, there is a need for an economical digital imaging system for general radiology. The x-ray light valve (XLV) is a novel digital x ray detector concept with the potential for high image quality and low cost. The XLV is comprised of a photoconductive detector layer and liquid crystal (LC) cell physically coupled in a sandwich structure. Upon exposure to x rays, charge is collected at the surface of the photoconductor, causing a change in the reflective properties of the LC cell. The visible image so formed can subsequently be digitized with an optical scanner. By choosing the properties of the LC cell in combination with the appropriate photoconductor thickness and bias potentials, the XLV can be optimized for various diagnostic imaging tasks. Specifically for chest radiography, we identified three potentially practical reflective cell designs by selecting from those commonly used in LC display technology. The relationship between reflectance and x-ray exposure (i.e., the characteristic curve) was determined for all three cells using a theoretical model. The results indicate that the reflective electrically controlled birefringence (r-ECB) cell is the preferred choice for chest radiography, provided that the characteristic curve can be shifted towards lower exposures. The feasibility of the shift of the characteristic curve is shown experimentally. The experimental results thus demonstrate that an XLV based on the r-ECB cell design exhibits a characteristic curve suitable for chest radiography. PMID- 18404933 TI - X-ray imaging performance of scintillator-filled silicon pore arrays. AB - The need for fine detail visibility in various applications such as dental imaging, mammography, but also neurology and cardiology, is the driver for intensive efforts in the development of new x-ray detectors. The spatial resolution of current scintillator layers is limited by optical diffusion. This limitation can be overcome by a pixelation, which prevents optical photons from crossing the interface between two neighboring pixels. In this work, an array of pores was etched in a silicon wafer with a pixel pitch of 50 microm. A very high aspect ratio was achieved with wall thicknesses of 4-7 microm and pore depths of about 400 microm. Subsequently, the pores were filled with Tl-doped cesium iodide (CsI:Tl) as a scintillator in a special process, which includes powder melting and solidification of the CsI. From the sample geometry and x-ray absorption measurement the pore fill grade was determined to be 75%. The scintillator-filled samples have a circular active area of 16 mm diameter. They are coupled with an optical sensor binned to the same pixel pitch in order to measure the x-ray imaging performance. The x-ray sensitivity, i.e., the light output per absorbed x ray dose, is found to be only 2.5%-4.5% of a commercial CsI-layer of similar thickness, thus very low. The efficiency of the pores to transport the generated light to the photodiode is estimated to be in the best case 6.5%. The modulation transfer function is 40% at 4 lp/mm and 10%-20% at 8 lp/mm. It is limited most likely by the optical gap between scintillator and sensor and by K-escape quanta. The detective quantum efficiency (DQE) is determined at different beam qualities and dose settings. The maximum DQE(0) is 0.28, while the x-ray absorption with the given thickness and fill factor is 0.57. High Swank noise is suspected to be the reason, mainly caused by optical scatter inside the CsI-filled pores. The results are compared to Monte Carlo simulations of the photon transport inside the pore array structure. In addition, some x-ray images of technical and anatomical phantoms are shown. This work shows that scintillator-filled pore arrays can provide x-ray imaging with high spatial resolution, but are not suitable in their current state for most of the applications in medical imaging, where increasing the x-ray doses cannot be tolerated. PMID- 18404934 TI - A generalized a priori dose uncertainty model of IMRT delivery. AB - Multileaf collimator-based intensity modulated radiation therapy (IMRT) is complex because each intensity modulated field consists of hundreds of subfields, each of which is associated with an intricate interplay of uncertainties. In this study, the authors have revised the previously introduced uncertainty model to provide an a priori accurate prediction of dose uncertainty during treatment planning in IMRT. In the previous model, the dose uncertainties were categorized into space-oriented dose uncertainty (SOU) and nonspace-oriented dose uncertainty (NOU). The revised model further divided the uncertainty sources into planning and delivery. SOU and NOU associated with a planning system were defined as inherent dose uncertainty. A convolution method with seven degrees of freedom was also newly applied to generalize the model for practical clinical cases. The model parameters were quantified through a set of measurements, accumulated routine quality assurance (QA) data, and peer-reviewed publications. The predicted uncertainty maps were compared with dose difference distributions between computations and 108 simple open-field measurements using a two dimensional diode array detector to verify the validity of the model parameters and robustness of the generalized model. To examine the applicability of the model to overall dose uncertainty prediction in IMRT, a retrospective analysis of QA measurements using the diode array detector for 32 clinical IM fields was also performed. A scatter diagram and a correlation coefficient were employed to investigate a correlation of the predicted dose uncertainty distribution with the dose discrepancy distribution between calculation and delivery. In addition, a gamma test was performed to correlate failed regions in dose verification with the dose uncertainty map. The quantified model parameters well correlated the predicted dose uncertainty with the probable dose difference between calculations and measurements. It was visually validated with the scatter diagrams. The average correlation coefficient between uncertainty and dose difference of 108 verification measurements was 0.80 +/- 0.04, indicating a strong linear correlation. In the clinical IM field studies, the dose uncertainty map mimicked the probable dose difference distribution. The average correlation coefficient between the overall dose uncertainty and the dose difference of 32 QA measurements (total 13 184 comparison points) was 0.75 +/- 0.07, which also indicated a strong linear correlation between them. The failed regions of the gamma test remarkably corresponded to relatively high dose uncertainty. In conclusion, the dose uncertainty map was able to highlight high dose uncertainty regions, where more care should be taken during the treatment plan. The a priori accurate prediction of dose uncertainty in IMRT will significantly improve the treatment plan evaluation process, thus improving the quality of radiation treatments. PMID- 18404935 TI - A practical global distortion correction method for an image intensifier based x ray fluoroscopy system. AB - X-ray images acquired on systems with image intensifiers (II) exhibit characteristic distortion which is due to both external and internal factors. The distortion is dependent on the orientation of the II, a fact particularly relevant to II's mounted on C arms which have several degrees of freedom of motion. Previous descriptions of distortion correction strategies have relied on a dense sampling of the C-arm orientation space, and as such have been limited mostly to a single arc of the primary angle, alpha. We present a new method which smooths the trajectories of the segmented vertices of the grid phantom as a function of a prior to solving the two-dimensional warping problem. It also shows that the same residual errors of distortion correction could be achieved without fitting the trajectories of the grid vertices, but instead applying the previously described global method of distortion correction, followed by directly smoothing the values of the polynomial coefficients as functions of the C-arm orientation parameters. When this technique was applied to a series of test images at arbitrary alpha, the root-mean-square (RMS) residual error was 0.22 pixels. The new method was extended to three degrees of freedom of the C-arm motion: the primary angle, alpha; the secondary angle, beta; and the source-to intensifier distance, lambda. Only 75 images were used to characterize the distortion for the following ranges: alpha, +/- 45 degrees (Deltaalpha = 22.5 degrees); beta, +/- 36 degrees (Deltabeta = 18 degrees); lambda, 98-118 cm (Deltalambda = 10 cm). When evaluated on a series of test images acquired at arbitrary (alpha, beta, lambda), the RMS residual error was 0.33 pixels. This method is targeted at applications such as guidance of catheter-based interventions and treatment planning for brachytherapy, which require distortion corrected images over a large range of C-arm orientations. PMID- 18404936 TI - 4D-CT lung motion estimation with deformable registration: quantification of motion nonlinearity and hysteresis. AB - In this article, our goal is twofold. First, we propose and compare two methods which process deformable registration to estimate patient specific lung and tumor displacements and deformation during free breathing from four-dimensional computed tomography (4D-CT) data. Second, we propose techniques to quantify the physiological parameters of motion nonlinearity and hysteresis. A Frechet distance-based criterion is introduced to measure the motion hysteresis. Experiments were conducted with 4D-CT data of five patients treated in radiotherapy for non-small cell lung cancer. The accuracy of deformation fields assessed against expert-selected landmarks was found to be within image voxel tolerance. The second method gave slightly better results in terms of accuracy and consistency, although the differences were not statistically significant between the two methods. Lung motion nonlinearity and hysteresis are patient specific, and vary across regions within the lung during respiration. For all patients, motion between end-exhale and end-inhale was well approximated with a straight line trajectory for the majority of lung points. Hysteresis was found to be globally correlated with trajectory length. The main limitation to the proposed method is that intensity-based deformable registration is dependent on image quality and image resolution. Another limitation is the absence of gold standard which makes the validation of the computed motion difficult. However, the proposed tools provide patient specific motion information which, in radiotherapy for example, can ease the definition of precise internal margins. In the future, the integration of physiological information from multiple patients could help to create a general lung atlas with different clinical applications. PMID- 18404937 TI - Patient dosimetry for hybrid MRI-radiotherapy systems. AB - A novel geometry has been proposed for a hybrid magnetic resonance imaging (MRI) linac system in which a 6 MV linac is mounted on the open end of a biplanar, low field (0.2 T) MRI magnet on a single gantry that is free to rotate around the patient. This geometry creates a scenario in which the magnetic field vector remains fixed with respect to the incident photon beam, but moves with respect to the patient as the gantry rotates. Other proposed geometries are characterized by a radiation source rotating about a fixed cylindrical magnet where the magnetic field vector remains fixed with respect to the patient. In this investigation we simulate the inherent dose distribution patterns within the two MRI-radiation source geometries using PENELOPE and EGSnrc Monte Carlo radiation transport codes with algorithms implemented to account for the magnetic field deflection of charged particles. Simulations are performed in phantoms and for clinically realistic situations. The novel geometry results in a net Lorentz force that remains fixed with respect to the patient (in the cranial-caudal direction) and results in a cumulative influence on dose distribution for a multiple beam treatment scenario. For a case where patient anatomy is reasonably homogeneous (brain plan), differences in dose compared to a conventional (no magnetic field) case are minimal for the novel geometry. In the case of a lung plan where the inhomogeneous patient anatomy allows for the magnetic field to have significant influence on charged particle transport, larger differences occur in a predictable manner. For a system using a fixed cylindrical geometry and higher magnetic field (1.5 T), differences from the case without a magnetic field are significantly greater. PMID- 18404939 TI - Digital radiography using amorphous selenium: photoconductively activated switch (PAS) readout system. AB - A new amorphous selenium (a-Se) digital radiography detector is introduced. The proposed detector generates a charge image in the a-Se layer in a conventional manner, which is stored on electrode pixels at the surface of the a-Se layer. A novel method, called photoconductively activated switch (PAS), is used to read out the latent x-ray charge image. The PAS readout method uses lateral photoconduction at the a-Se surface which is a revolutionary modification of the bulk photoinduced discharge (PID) methods. The PAS method addresses and eliminates the fundamental weaknesses of the PID methods--long readout times and high readout noise--while maintaining the structural simplicity and high resolution for which PID optical readout systems are noted. The photoconduction properties of the a-Se surface were investigated and the geometrical design for the electrode pixels for a PAS radiography system was determined. This design was implemented in a single pixel PAS evaluation system. The results show that the PAS x-ray induced output charge signal was reproducible and depended linearly on the x-ray exposure in the diagnostic exposure range. Furthermore, the readout was reasonably rapid (10 ms for pixel discharge). The proposed detector allows readout of half a pixel row at a time (odd pixels followed by even pixels), thus permitting the readout of a complete image in 30 s for a 40 cm x 40 cm detector with the potential of reducing that time by using greater readout light intensity. This demonstrates that a-Se based x-ray detectors using photoconductively activated switches could form a basis for a practical integrated digital radiography system. PMID- 18404938 TI - Experimental verification and clinical implementation of a commercial Monte Carlo electron beam dose calculation algorithm. AB - This study describes the modeling and the experimental verification and clinical implementation of the alpha release of Pinnacle3 Monte Carlo (MC) electron beam dose calculation algorithm for patient-specific treatment planning. The MC electron beam modeling was performed for beam energies ranging from 6 to 18 MeV from a Siemens (Primus) linear accelerator using standard-shaped electron applicators and 100 cm source-to-surface distance (SSD). The agreement between MC calculations and measurements was, on average, within 2% and 2 mm for all applicator sizes. However, differences of the order of 3%-4% were noted in the off-axis dose profiles for the largest applicator modeled and for all energies. Output factors were calculated for standard electron cones and square cutouts inserted in the 10 x 10 cm2 applicator for different SSDs and were found to be within 4% of measured data. Experimental verification of the MC electron beam model was carried out using an ionization chamber and film in solid-water slab and anthropomorphic phantoms containing bone and lung materials. Agreement between calculated and measured dose distributions was within +/-3%. Clinical comparison was performed in four patient treatment plans with lesions in highly irregular anatomies, such as the ear, face, and breast, where custom-designed bolus and field shaping blocks were used in the patient treatments. For comparison purposes, treatment planning was also performed using the conventional pencil beam (PB) algorithm with the Pinnacle3 treatment planning system. Differences between MC and PB dose calculations for the patient treatment plans were significant, particularly in anatomies where the target was in close proximity to low density tissues, such as lung and air cavities. Concerning monitor unit calculations, the largest differences obtained between MC and PB algorithms were between 4.0% and 5.0% for two patients treated with oblique beams and involving highly irregular surfaces, i.e., breast and cheek. Clinical results are reported for overall uncertainty values (averaged over voxels with doses >50% dosemax) ranging from 2% to 0.3% and calculations were performed using cubic voxels with side 0.3 cm. Timing values ranged from 2 min to 24.5 h, depending on the field size, beam energy, number, and thickness of computed tomography slices used to define the patient's anatomy for the overall uncertainty values mentioned above. PMID- 18404941 TI - Antialiasing backprojection for helical MDCT. AB - Helical CTs are well known to suffer from aliasing artifacts because of their finite longitudinal sampling pitch. The artifact pattern is typically strong streaks from bone edges in clinical images. Especially in the case of multidetector row CT, the artifact resulting from longitudinal aliasing is often called a windmill artifact because the visible streaks form a windmill pattern when the object is of a particular shape. The scan must be performed using a very thin slice thickness, i.e., fine sampling in the longitudinal direction, with a longer scan time to mitigate this aliasing artifact. Some elaborate longitudinal interpolation methods to remediate longitudinal aliasing have been proposed, but they have not been successful in practice despite their theoretical importance. A periodic swing of the focal spot in the longitudinal direction, a so-called z flying focal spot, was introduced recently to achieve finer sampling. Although it is a useful technique, some important deficiencies exist: It is sufficiently effective only near the isocenter and is difficult to apply to a scan using a thick slice thickness, even though longitudinal aliasing is more serious at the thicker scan. In this paper, the author addresses the nature of interlaced (or unequally spaced) sampling and derives a new principle of data treatment that can suppress the aliased spectra selectively. According to this principle, the common practice of image reconstruction, which backprojects data along the original sampling ray path, is never the best choice. The author proposes a new scheme of backprojection, which involves the longitudinal shift of projection data. A proper choice of longitudinal shift for backprojection provides effective and selective suppression of aliased spectra, with retention of the original frequency spectrum depending on the level of focus swing. With this shifted backprojection, the swing of focus can be made much smaller than for a conventional z-flying focal spot. The required amount of shift for backprojection is position dependent. Nevertheless, its implementation in the reconstruction process can be achieved simply by relocating the x-ray source and detector assembly from positions of actual scanning. Through simulation, the combination of shifted backprojection and the small swing of focus is evaluated. Results confirm that the artifact attributable to longitudinal aliasing is well suppressed in the entire field of view, whereas the penalty on the slice sensitivity profile (or longitudinal resolution) can be kept minimal. Moreover, this method solves other deficiencies of z-flying focus, such as inapplicability to scans with a thicker slice thickness. PMID- 18404940 TI - An outlook on x-ray CT research and development. AB - Over the past decade, computed tomography (CT) theory, techniques and applications have undergone a rapid development. Since CT is so practical and useful, undoubtedly CT technology will continue advancing biomedical and non biomedical applications. In this outlook article, we share our opinions on the research and development in this field, emphasizing 12 topics we expect to be critical in the next decade: analytic reconstruction, iterative reconstruction, local/interior reconstruction, flat-panel based CT, dual-source CT, multi-source CT, novel scanning modes, energy-sensitive CT, nano-CT, artifact reduction, modality fusion, and phase-contrast CT. We also sketch several representative biomedical applications. PMID- 18404943 TI - Evaluation of residual patient position variation for spinal radiosurgery using the Novalis image guided system. AB - PURPOSE: The Novalis system has been demonstrated to achieve accurate target localization on anthropomorphic phantoms. However, other factors, such as rotational deviation, patient intrafraction motion, and image fusion uncertainty due to patient body deformation, could contribute additional position uncertainty for actual patients. This study evaluates such position uncertainty for spinal radiosurgery patients. MATERIALS AND METHODS: Fifty-two consecutive spinal radiosurgery patients were included in the study. Rotational deviation was evaluated from 6-deg of freedom (6D) fusion results for all patients. The combined uncertainty of patient motion and image fusion was determined from fusion results of additional kV x-ray images acquired before, during, and after treatment for 25 of the 52 patients. The uncertainty of image fusion was also evaluated by performing 6D fusion ten different times with various regions of interest in the images selected for fusion. This was performed for two patients with L3 and T2 lesions, respectively, for comparison. RESULTS: The mean rotational deviation was 0.7 +/- 1.8, 0.7 +/- 1.5, and 0.7 +/- 1.6 deg along the yaw, roll, and pitch directions, respectively. The combined uncertainty from patient motion and image fusion was 0.1 +/- 0.9, 0.2 +/- 1.2, and 0.2 +/- 1.0 mm in the anteroposterior (AP), longitudinal, and lateral directions, respectively. The uncertainty (standard deviation) due to image fusion was less than 0.28 mm in any direction for the L3 lesion and 0.8 mm in the AP direction for the T2 lesion. CONCLUSION: Overall position uncertainty for spinal radiosurgery patients has been evaluated. Rotational deviation and patient motion were the main factors contributed to position uncertainty for actual patient treatment. PMID- 18404942 TI - Classification of breast computed tomography data. AB - Differences in breast tissue composition are important determinants in assessing risk, identifying disease in images and following changes over time. This paper presents an algorithm for tissue classification that separates breast tissue into its three primary constituents of skin, fat and glandular tissue. We have designed and built a dedicated breast CT scanner. Fifty-five normal volunteers and patients with mammographically identified breast lesions were scanned. Breast CT voxel data were filtered using a 5 pt median filter and the image histogram was computed. A two compartment Gaussian fit of histogram data was used to provide an initial estimate of tissue compartments. After histogram analysis, data were input to region-growing algorithms and classified as to belonging to skin, fat or gland based on their value and architectural features. Once tissues were classified, a more detailed analysis of glandular tissue patterns and a more quantitative analysis of breast composition was made. Algorithm performance assessment demonstrated very good or excellent agreement between algorithm and radiologist observers in 97.7% of the segmented data. We observed that even in dense breasts the fraction of glandular tissue seldom exceeded 50%. For most individuals the composition is better characterized as being a 70% (fat)-30% (gland) composition than a 50% (fat)-50% (gland) composition. PMID- 18404944 TI - A deformable phantom for 4D radiotherapy verification: design and image registration evaluation. AB - Motion of thoracic tumors with respiration presents a challenge for three dimensional (3D) conformal radiation therapy treatment. Validation of techniques aimed at measuring and minimizing the effects of respiratory motion requires a realistic deformable phantom for use as a gold standard. The purpose of this study was to develop and study the characteristics of a reproducible, tissue equivalent, deformable lung phantom. The phantom consists of a Lucite cylinder filled with water containing a latex balloon stuffed with dampened natural sponges. The balloon is attached to a piston that mimics the human diaphragm. Nylon wires and Lucite beads, emulating vascular and bronchial bifurcations, were uniformly glued at various locations throughout the sponges. The phantom is capable of simulating programmed irregular breathing patterns with varying periods and amplitudes. A tissue equivalent tumor, suitable for holding radiochromic film for dose measurements was embedded in the sponge. To assess phantom motion, eight 3D computed tomography data sets of the static phantom were acquired for eight equally spaced positions of the piston. The 3D trajectories of 12 manually chosen point landmarks and the tumor center-of-mass were studied. Motion reproducibility tests of the deformed phantom were established on seven repeat scans of three different states of compression. Deformable image registration (DIR) of the extreme breathing phases was performed. The accuracy of the DIR was evaluated by visual inspection of image overlays and quantified by the distance-to-agreement (DTA) of manually chosen point landmarks and triangulated surfaces obtained from 3D contoured structures. In initial tests of the phantom, a 20-mm excursion of the piston resulted in deformations of the balloon of 20 mm superior-inferior, 4 mm anterior-posterior, and 5 mm left-right. The change in the phantom mean lung density ranged from 0.24 (0.12 SD) g/cm3 at peak exhale to 0.19 (0.12 SD) g/cm3 at peak inhale. The SI displacement of the landmarks varied between 94% and 3% of the piston excursion for positions closer and farther away from the piston, respectively. The reproducibility of the phantom deformation was within the image resolution (0.7 x 0.7 x 1.25 mm3). Vector average registration accuracy based on point landmarks was found to be 0.5 (0.4 SD) mm. The tumor and lung mean 3D DTA obtained from triangulated surfaces were 0.4 (0.1 SD) mm and 1.0 (0.8 SD) mm, respectively. This phantom is capable of reproducibly emulating the physically realistic lung features and deformations and has a wide range of potential applications, including four-dimensional (4D) imaging, evaluation of deformable registration accuracy, 4D planning and dose delivery. PMID- 18404945 TI - A computational implementation and comparison of several intensity modulated proton therapy treatment planning algorithms. AB - The authors present a comparative study of intensity modulated proton therapy (IMPT) treatment planning employing algorithms of three-dimensional (3D) modulation, and 2.5-dimensional (2.5D) modulation, and intensity modulated distal edge tracking (DET) [A. Lomax, Phys. Med. Biol. 44, 185-205 (1999)] applied to the treatment of head-and-neck cancer radiotherapy. These three approaches were also compared with 6 MV photon intensity modulated radiation therapy (IMRT). All algorithms were implemented in the University of Florida Optimized Radiation Therapy system using a finite sized pencil beam dose model and a convex fluence map optimization model. The 3D IMPT and the DET algorithms showed considerable advantages over the photon IMRT in terms of dose conformity and sparing of organs at risk when the beam number was not constrained. The 2.5D algorithm did not show an advantage over the photon IMRT except in the dose reduction to the distant healthy tissues, which is inherent in proton beam delivery. The influences of proton beam number and pencil beam size on the IMPT plan quality were also studied. Out of 24 cases studied, three cases could be adequately planned with one beam and 12 cases could be adequately planned with two beams, but the dose uniformity was often marginally acceptable. Adding one or two more beams in each case dramatically improved the dose uniformity. The finite pencil beam size had more influence on the plan quality of the 2.5D and DET algorithms than that of the 3D IMPT. To obtain a satisfactory plan quality, a 0.5 cm pencil beam size was required for the 3D IMPT and a 0.3 cm size was required for the 2.5D and the DET algorithms. Delivery of the IMPT plans produced in this study would require a proton beam spot scanning technique that has yet to be developed clinically. PMID- 18404946 TI - A comparison of neural network approaches for on-line prediction in IGRT. AB - Image-guided radiation therapy aims to improve the accuracy of treatment delivery by tracking tumor position and compensating for observed movement. Due to system latency it is sometimes necessary to predict tumor trajectory evolution in order to facilitate changes in beam delivery. Neural networks (NNs) have previously been investigated for predicting future tumor position because of their ability to model non-linear systems. However, no attempt has been made to optimize the NN training algorithms, and no mention has been made of potential errors which can be caused by using NNs for extrapolation purposes. In this work, after giving a brief explanation of NN theory, a comparison is made between 4 different adaptive algorithms for training time-series prediction NNs. New error criteria are introduced which highlight error maxima. Results are obtained by training the NNs using previously published data. A hybrid algorithm combining Bayesian regularization with conjugate-gradient backpropagation is demonstrated to give the best average prediction accuracy, whilst a generalized regression NN is shown to reduce the possibility of isolated large prediction errors. PMID- 18404947 TI - Extension of the biological effective dose to the MIRD schema and possible implications in radionuclide therapy dosimetry. AB - In dosimetry-based treatment planning protocols, patients with rapid clearance of the radiopharmaceutical require a larger amount of initial activity than those with slow clearance to match the absorbed dose to the critical organ. As a result, the dose-rate to the critical organ is higher in patients with rapid clearance and may cause unexpected toxicity compared to patients with slow clearance. In order to account for the biological impact of different dose-rates, radiobiological modeling is beginning to be applied to the analysis of radionuclide therapy patient data. To date, the formalism used for these analyses is based on kinetics derived from activity in a single organ, the target. This does not include the influence of other source organs to the dose and dose-rate to the target organ. As a result, only self-dose irradiation in the target organ contributes to the dose-rate. In this work, the biological effective dose (BED) formalism has been extended to include the effect of multiple source organ contributions to the net dose-rate in a target organ. The generalized BED derivation has been based on the Medical Internal Radionuclide Dose Committee (MIRD) schema assuming multiple source organs following exponential effective clearance of the radionuclide. A BED-based approach to determine the largest safe dose to critical organs has also been developed. The extended BED formalism is applied to red marrow dosimetry, as well as kidney dosimetry considering the cortex and the medulla separately, since both those organs are commonly dose limiting in radionuclide therapy. The analysis shows that because the red marrow is an early responding tissue (high alpha/beta), it is less susceptible to unexpected toxicity arising from rapid clearance of high levels of administered activity in the marrow or in the remainder of the body. In kidney dosimetry, the study demonstrates a complex interplay between clearance of activity in the cortex and the medulla, as well as the initial activity ratio and the S value ratio between the two. In some scenarios, projected BED based on both the cortex and the medulla is a more appropriate constraint on the administered activity than the BED based on the cortex only. Furthermore, different fractionated regimens were considered to reduce renal toxicity. The MIRD-based BED formalism is expected to be useful for patient-specific adjustments of activity and to facilitate the investigation of dose-toxicity correlations with respect to dose rate and tissue repair mechanism. PMID- 18404948 TI - Accurate patient dosimetry of kilovoltage cone-beam CT in radiation therapy. AB - The increased utilization of x-ray imaging in image-guided radiotherapy has dramatically improved the radiation treatment and the lives of cancer patients. Daily imaging procedures, such as cone-beam computed tomography (CBCT), for patient setup may significantly increase the dose to the patient's normal tissues. This study investigates the dosimetry from a kilovoltage (kV) CBCT for real patient geometries. Monte Carlo simulations were used to study the kV beams from a Varian on-board imager integrated into the Trilogy accelerator. The Monte Carlo calculated results were benchmarked against measurements and good agreement was obtained. The authors developed a novel method to calibrate Monte Carlo simulated beams with measurements using an ionization chamber in which the air kerma calibration factors are obtained from an Accredited Dosimetry Calibration Laboratory. The authors have introduced a new Monte Carlo calibration factor, fMCcal, which is determined from the calibration procedure. The accuracy of the new method was validated by experiment. When a Monte Carlo simulated beam has been calibrated, the simulated beam can be used to accurately predict absolute dose distributions in the irradiated media. Using this method the authors calculated dose distributions to patient anatomies from a typical CBCT acquisition for different treatment sites, such as head and neck, lung, and pelvis. Their results have shown that, from a typical head and neck CBCT, doses to soft tissues, such as eye, spinal cord, and brain can be up to 8, 6, and 5 cGy, respectively. The dose to the bone, due to the photoelectric effect, can be as much as 25 cGy, about three times the dose to the soft tissue. The study provides detailed information on the additional doses to the normal tissues of a patient from a typical kV CBCT acquisition. The methodology of the Monte Carlo beam calibration developed and introduced in this study allows the user to calculate both relative and absolute absorbed doses. PMID- 18404949 TI - How does CT image noise affect 3D deformable image registration for image-guided radiotherapy planning? AB - PURPOSE: To measure the sensitivity of deformable image registration to image noise. Deformable image registration can be used to map organ contours and other treatment planning data from one CT to another. These CT studies can be acquired with either conventional fan-beam CT systems or more novel cone-beam CT techniques. However, cone-beam CT images can have higher noise levels than fan beam CT, which might reduce registration accuracy. We have investigated the effect of image quality differences on the deformable registration of fan-beam CTs and CTs with simulated cone-beam noise. METHOD: Our study used three CT studies for each of five prostate patients. Each CT was contoured by three experienced radiation oncologists. For each patient, one CT was designated the source image and the other two were target images. A deformable image registration process was used to register each source CT to each target CT and then transfer the manually drawn treatment planning contours from the source CT to the target CTs. The accuracy of the automatically transferred contours (and thus of the deformable registration process) was assessed by comparing them to the manual contours on the target CTs, with the differences evaluated with respect to interobserver variability in the manual contours. Then each of the target CTs was modified to include increased noise characteristic of cone-beam CT and the tests were repeated. Changes in registration accuracy due to increased noise were detected by monitoring changes in the automatically transferred contours. RESULTS: We found that the additional noise caused no significant loss of registration accuracy at magnitudes that exceeded what would normally be found in an actual cone-beam CT. SUMMARY: We conclude that noise levels in cone-beam CTs that might reduce manual contouring accuracy do not reduce image registration and automatic contouring accuracy. PMID- 18404950 TI - Dosimetric characterization of Ir-192 LDR elongated sources. AB - Ir-192 wires have been used in low-dose-rate brachytherapy for many years. Commercially available treatment planning systems approximate the dose rate distribution of the straight or curved wires applying the superposition principle using one of the following methods: (i) The wire is modeled as a set of point sources, (ii) the wire is modeled as a set of small straight segment wires, (iii) the values of the parameters and functions of the American Association of Physicists in Medicine (AAPM) Task Group 43 protocol are obtained for wire lengths between 3 and 7 cm assuming some simplifications. The dose rate distributions obtained using these methods for linear wires of different lengths and U-shaped wires present significant deviations compared to those obtained by Monte Carlo. In the present study we propose a new method to model 192Ir wires of any length and shape, named the Two Lengths based Segmented method. This method uses the formalism stated in the AAPM Task Group 43 protocol for two straight wires only, 0.5 and 1 cm, to obtain the dose rate distribution around wires of any length (down to 0.3 cm and up to 10 cm) improving on the results of the aforementioned ones. This method can easily be applied to dose calculations around other wires, such as Pd-103 ones. PMID- 18404951 TI - Conjunctivitis. PMID- 18404952 TI - What's your diagnosis? Juvenile xanthogranuloma of the iris. PMID- 18404953 TI - Pediatric ophthalmologic consultations in a children's hospital setting. PMID- 18404954 TI - Ocular infections due to anaerobic bacteria in children. AB - The recent increased recovery of anaerobic bacteria from children has led to greater appreciation of the role these organisms play in pediatric infections at all body sites, including the eye. In studies that employed adequate methods for recovery of anaerobes, they were isolated from approximately one-third of the children with conjunctivitis, half of the time in pure culture. The predominant recovered anaerobes were Clostridium spp., gram-negative anaerobic bacilli, and Peptostreptococcus spp. Anaerobic bacteria were also recovered from patients who wore contact lenses and developed conjunctivitis, and were reported in cases of keratitis. The most frequently recovered anaerobes were Propionibacterium spp., Peptostreptococcus spp., Clostridium spp., Prevotella spp., and Fusobacterium spp. The most frequently recovered anaerobes from dacryocystitis were Peptostreptococcus spp., Propionibacterium spp., Prevotella spp., and Fusobacterium spp. This review describes the microbiology, diagnosis, and management of ocular infections due to anaerobic bacteria in children. PMID- 18404955 TI - The frequency of ocular abnormalities in inpatient pediatric ophthalmology consultations. AB - OBJECTIVE: To determine the frequency, type, and results of pediatric ophthalmology service consultations at a tertiary care children's hospital and to offer advice as to the timing of the initial consultation based on the patient's diagnosis and likelihood of ocular disease, METHOD: A retrospective evaluation of inpatient ophthalmology consultations from September 1, 2003, to August 31, 2004, at Texas Children's Hospital was conducted. Patients were identified using the Current Procedural Terminology listing of billing codes for various levels of service for new initial inpatient consultations. RESULTS: During the 1-year period, 445 new inpatient consultations were requested from the pediatric ophthalmology service, primarily to rule out ophthalmic problems or manifestations (55.9%) and to evaluate ocular complaints or ocular abnormalities as noted by the primary team (44.1%). Of the 445 patients, 215 (48.3%) were found to have ocular abnormalities and 230 (51.7%) had no ocular abnormalities at the time of initial consultation. CONCLUSION: Patients with ocular signs or symptoms of disease should receive urgent ophthalmic consultation. Consideration should be given to the usefulness of urgent consultations in patients suspected of having fungemia, sepsis, and headache. PMID- 18404956 TI - Higher order aberrations in children with amblyopia. AB - PURPOSE: To quantify and characterize higher order aberrations in eyes of children with amblyopia. METHODS: Higher order aberrations were measured in a group of children using the iTrace Visual Function Analyzer (Tracey Technologies, Houston, TX). Children involved in the study had unilateral amblyopia secondary to strabismus or anisometropia. RESULTS: Thirty children between 6 and 17 years old (mean: 10.7 years) were studied. Fifteen children had strabismic and 15 had anisometropic amblyopia. No statistically significant difference was found in mean root mean square values of total higher order aberrations, coma, spherical, higher order astigmatism, trefoil, or 3rd, 4th, 5th, or 6th order terms of normal and amblyopic eyes. CONCLUSION: Higher order aberrations are similar in amblyopic eyes due to strabismus or anisometropia and normal fellow eyes. Unlike lower order aberrations such as sphere and cylinder, higher order aberrations are unlikely to play a role in the development of amblyopia. PMID- 18404957 TI - A pilot study of the value of prism adaptation in planning strabismus reoperations. AB - PURPOSE: To evaluate the value of prism adaptation in planning strabismus reoperations. METHODS: Of 15 patients with persistent strabismus, 9 underwent prism adaptation prior to strabismus reoperation and 6 underwent strabismus reoperation without preoperative prism adaptation. Corrective surgery was performed on the same or new muscles. Follow-up ranged from 3 months to 2 years. A horizontal deviation of 10 prism diopters or less was considered a successful outcome. Statistical analysis of the results was done. RESULTS: Of the 9 patients who had prism adaptation, 3 (33%) were responders and 6 (67%) were nonresponders. A successful outcome was achieved in 6 patients (67%) who had prism adaptation and 3 patients (50%) who did not. The difference between groups was statistically insignificant (P > .05). CONCLUSION: These results suggest that the benefits of prism adaptation are not significant enough to justify the time consumption and expenses of this test when planning strabismus reoperations, but a larger group of patients should be studied. PMID- 18404958 TI - Binocular potential score: a novel concept. AB - OBJECTIVE: To assess the predictive value of an objective system for preoperative binocular potential scoring on the postoperative outcome in horizontal concomitant strabismus. METHODS: A prospective interventional study of 100 patients undergoing surgery for horizontal concomitant strabismus was conducted. The binocular potential score (BPS) was evaluated on the basis of age of onset, duration, intermittency, variability, vision, and responses on synoptophore and Worth four-dot test. Patients were grouped according four grades (I = the best and IV = the weakest). The surgical outcome was evaluated by binocular function and ocular alignment. RESULTS: All patients with a BPS of grade I maintained good binocular function postoperatively. Within-grade analysis revealed a statistically significant improvement in postoperative binocular function in patients with a BPS of grade II (P = .0047), grade III (P = .0030), and grade IV (P = .0143). Grade comparisons showed significant differences between grades II and IV (P = .00) and grades III and IV (P = .0005), but not between grades II and III. CONCLUSIONS: The BPS is a useful tool for predicting surgical outcome and it may be valuable to conduct multicentric trials using this objective measurement. PMID- 18404959 TI - Fluorescein angiography of iris juvenile xanthogranuloma. AB - A 3-year-old boy was referred because of the presence of a non-pigmented, highly vascular mass with tapioca appearance involving the superior portion of the left iris. Iris fluorescein angiography revealed early hyperfluorescence of the iris tumor with diffuse, intense late leakage of dye throughout the entire iris, not just in the region of the tumor. Cytopathologic examination revealed Touton giant cells and the presence of histiocytes, confirming the diagnosis of iris juvenile xanthogranuloma. The patient was prescribed topical prednisolone acetate, and the lesion resolved during the next 3 months. Fluorescein angiography may be useful in differentiating iris juvenile xanthogranuloma from malignant or non inflammatory benign iris lesions. PMID- 18404960 TI - Rhabdomyoma of the orbit. AB - The authors describe a rare case of orbital rhabdomyoma in a 3-year-old girl who presented with progressive proptosis of the left eye. An axial computed tomographic scan of the left orbit demonstrated an irregular retrobulbar mass. The tumor was resected locally from the lateral wall of the orbit and the resected specimens were diagnosed as orbital rhabdomyoma. The authors review the literature and discuss the diagnostic implications and treatment strategies. PMID- 18404961 TI - Ring corneal infiltrate and progressive ring thinning following primary varicella infection. AB - Unilateral stromal keratitis is a known rare sequela of primary varicella infection. The authors describe a unique case of immunological (Wessely) ring formation and progressive ring thinning following primary varicella infection in a 6-year-old girl. PMID- 18404962 TI - Retinopathy of prematurity treatment and vitreous hemorrhage. AB - Two cases of hemorrhagic retinopathy of prematurity associated with thrombocytopenia are presented. The negative prognosis of vitreous hemorrhage in retinopathy of prematurity and the option of platelet transfusions are discussed. PMID- 18404963 TI - Multiple pilomatrixomas affecting the eyelid and face. AB - A 3-year-old girl presented with a right upper eyelid mass. The lesion had a reddish appearance, was firm to palpation, and was fixed to underlying tissues. Fourteen months after the lesion was excised, a similar lesion was discovered on the left side of the nose and was also excised. Histopathologic examination of the excised tumors revealed variable basophilic hair matrix cells and sheets of nonviable eosinophilic shadow cells. Foci of dystrophic calcification were also seen in the necrotic tumor areas. The histopathologic findings were found to be consistent with pilomatrixoma. Results of limited clinical work-up of the child for Gardner's syndrome, sarcoidosis, and myotonic dystrophy were negative. Multiple periocular and facial pilomatrixomas can occur in children in the clinical absence of myotonic dystrophy, Gardner's syndrome, and sarcoidosis. PMID- 18404964 TI - Clinical evolution of neuroretinitis in Parry-Romberg syndrome. AB - A 16-year-old girl with Parry-Romberg syndrome presented with monolateral exudative neuroretinitis and retinal telangiectasis that had been observed for 42 months. She was treated with immunosuppressive therapy with A-cyclosporine for 1 year, followed by laser treatment of telangiectasis. Her visual acuity improved from 20/200 to 20/70 in 1 year with a reduction of neuroretinal exudation. Laser treatment resulted in a further improvement of visual acuity to 20/40. Clinical findings remained unchanged during 2 years of follow-up. The association of immunosuppressive treatment and laser therapy may improve neuroretinal disease in the presence of Parry-Romberg syndrome. PMID- 18404965 TI - The early seeds of obesity: are childhood obesity programs too late to the table? PMID- 18404966 TI - Adolescent's refusal of treatment: principles in conflict. PMID- 18404967 TI - Hyperpigmentation in only one identical twin. PMID- 18404968 TI - Impact of early postnatal weight gain and feeding patterns on body mass index in adolescence. AB - Postnatal weight gain may predispose to the development of obesity during childhood. The aims of this study were to study the impact of weight gain during specific periods of the first year of life and of feeding patterns on the body mass index (BMI) of adolescents. Growth records during the first 24 months of life of 88 obese and 214 non-obese 12 year-old Arab children were evaluated. Birth weight and length were similar for obese and non-obese adolescents, while the rate of breastfeeding was significantly lower in the obese group (p < 0.01). Obese adolescents demonstrated a small yet significant accelerated weight gain at 4 (p = 0.002) and 12 (p = 0.01) months of age. Weight gain during the first 2 months of life and feeding pattern were independent predictors of BMI at the age of 12 years. Thus, early postnatal weight gain is associated with obesity in adolescence, while breastfeeding seems to have a protective effect. PMID- 18404970 TI - Serum adiponectin, C-reactive protein and TNF-alpha levels in obese Korean children. AB - AIM: To determine whether there are differences of serum adiponectin, CRP and TNF a levels between obese and non-obese children and to investigate the relationship between childhood obesity and inflammation. METHODS: A total of 103 (73 obese, 30 non-obese) children were evaluated for height, weight, blood pressure and body mass index (BMI). We measured serum adiponectin, hs-CRP, and TNF-alpha levels and analyzed their correlation with BMI, serum glucose, lipid profile and insulin resistance. RESULTS: Serum adiponectin levels were significantly decreased in the obese children compared to non-obese children, and had a negative correlation with BMI, fasting glucose and HOMA-IR. Serum hs-CRP and TNF-alpha levels were increased in obese children compared to non-obese children, and had a positive correlation with BMI. CONCLUSIONS: Serum adiponectin level was decreased and serum CRP and TNF-alpha levels were increased in obese children. Our findings suggest that childhood obesity is associated with inflammatory processes. PMID- 18404969 TI - Leptin, insulin, and glucose serum levels in large-for-gestational-age infants of diabetic and non-diabetic mothers. AB - BACKGROUND: It has been suggested that hyperleptinemia could be caused by hyperinsulinemia in infants of diabetic mothers (IDMs). AIM: To compare leptin, insulin, and glucose levels in large-for-gestational-age (LGA) and appropriate for-gestational-age (AGA) infants. METHODS: A cross-sectional study was conducted in IDMs, infants of non-diabetic mothers (INDM) and AGA infants. RESULTS: Seventy seven newborns were studied (11 IDM, 16 INDM, and 50 AGA infants). Leptin levels were significantly higher in LGA infants than in the AGA group and a trend for higher levels in IDM than NIDM was observed. Insulin levels and insulin resistance were significantly higher in IDMs. Glucose levels were lower in both groups of LGA infants. CONCLUSIONS: We found insulin resistance, hyperinsulinism and hyperleptinemia in IDMs, and the trend of higher leptin levels in IDMs than INDMs shows that leptin could be related to insulin resistance in these infants. PMID- 18404971 TI - Glucose fluxes during OGTT in adolescents assessed by a stable isotope triple tracer method. AB - Virtually no information is available on glucose fluxes during a meal or glucose ingestion in adolescents. AIM: To use a triple tracer approach to measure rates of appearance of ingested glucose (Ra(ogtt)), endogenous glucose production (EGP) and glucose disappearance (Rd) following an oral glucose bolus in adolescents. METHODS: Eleven adolescents (4 M/7 F, 15 +/- 1 yr; 67.3 +/- 4.7 kg; 24 +/-2 kg/m2) underwent a frequent sampled oral glucose tolerance test (OGTT) (labelled with [6,6-2H2]glucose) combined with intravenous infusion of [1-(13)C]glucose and [U-(13)C6]glucose following an overnight fast. Formulas were developed to estimate glucose fluxes using one- or two-compartment models. RESULTS: During the 7 h following the OGTT bolus, 9.8 +/- 2.3% of the ingested glucose was extracted by the liver, EGP was suppressed by 45 +/- 4% and Rd increased by 21 +/- 5%. CONCLUSIONS: The triple tracer method provided accurate assessment of Ra(ogtt), EGP and Rd fluxes during an OGTT in adolescents. Thus, this method might provide novel insight on postprandial glucose fluxes in children/adolescents under various conditions. PMID- 18404972 TI - Novel splice site mutation in the growth hormone receptor gene in Turkish patients with Laron-type dwarfism. AB - Growth hormone (GH) is involved in growth, and fat and carbohydrate metabolism. Interaction of GH with the GH receptor (GHR) is necessary for systemic and local production of insulin-like growth factor-I (IGF-I) which mediates GH actions. Mutations in the GHR cause severe postnatal growth failure; the disorder is an autosomal recessive genetic disease resulting in GH insensitivity, called Laron syndrome. It is characterized by dwarfism with elevated serum GH and low levels of IGF-I. We analyzed the GHR gene for mutations and polymorphisms in eight patients with Laron-type dwarfism from six families. We found three missense mutations (S40L, V125A, I526L), one nonsense mutation (W157X), and one splice site mutation in the extracellular domain of GHR. Furthermore, G168G and exon 3 deletion polymorphisms were detected in patients with Laron syndrome. The splice site mutation, which is a novel mutation, was located at the donor splice site of exon 2/ intron 2 within GHR. Although this mutation changed the highly conserved donor splice site consensus sequence GT to GGT by insertion of a G residue, the intron splicing between exon 2 and exon 3 was detected in the patient. These results imply that the splicing occurs arthe GT site in intron 2, leaving the extra inserted G residue at the end of exon 2, thus changing the open reading frame of GHR resulting in a premature termination codon in exon 3. PMID- 18404973 TI - ICR1 epimutations in llp15 are restricted to patients with Silver-Russell syndrome features. AB - (Epi)mutations affecting chromosome llp15 are well known to be associated with growth disturbances. The finding of llp15 mutations in the overgrowth disease Beckwith-Wiedemann syndrome led to the identification of imprinted growth promoting genes which are expressed paternally and of imprinted growth suppressing genes in the same region that are expressed maternally. An opposite epimutation in the same region is associated with Silver-Russell syndrome (SRS), a growth retardation disorder characterised by a typical facial gestalt, clinodactyly V and asymmetry. In more than 30% of patients with SRS, hypomethylation of the telomeric llp15 imprinting domain (ICR1) can be detected. However, the general significance of this epimutation for human growth retardation was unclear. In a previous study' we showed that the ICR1 epimutation is not present in growth retarded patients with dysmorphisms not typical for SRS, but its role in the development of isolated growth restriction needed to be further elucidated. We therefore screened 30 patients with isolated pre- and postnatal growth retardation (IUGR/PNGR) and 65 patients diagnosed with SRS by external clinicians for ICR1I epimutations. In the latter group clinical data were rarely provided. These 65 'SRS' patients were additionally analysed for maternal uniparental disomy 7 (matUPD7). We excluded ICR1 hypomethylation in all 30 patients with isolated growth retardation. In the SRS group, we detected four cases with ICR1 epimutation and three with matUPD7. By combining our data with those from our previous study we could show that the hypomethylation of ICR1 in llp15 is indeed restricted to patients with SRS features and can be disregarded in isolated IUGR/PNGR. Thus, testing for the epimutation is indicated only in case of growth restriction in association with clinical signs reminiscent of SRS. The low detection rate of the ICR1 epimutation in our 'SRS' group can be explained by the clinical heterogeneity of cases referred by external institutions. PMID- 18404974 TI - Cyclic pamidronate therapy in children with osteogenesis imperfecta: results of treatment and follow-up after discontinuation. AB - BACKGROUND: Cyclic intravenous pamidronate treatment is widely used for symptomatic therapy of osteogenesis imperfecta (OI). However, data after discontinuation are very limited. AIM: The results of cyclical pamidronate treatment in 14 patients with moderate/severe OI and follow up of six of them after discontinuation are presented to assess the effects of pamidronate and its discontinuation. PATIENTS AND METHODS: Pamidronate was administered at a dosage of 0.5 mg/kg for 3 successive days every 2 months in 14 patients with OI aged 5.10 +/- 3.68 years. Treatment was stopped in six patients after a duration of 16.33 +/- 4.63 months, due to stable bone mineral density (BMD) values and/or no fracture in the last 6 months, or due to family demand. The main outcome measures were areal BMD (aBMD) of the lumbar spine, biochemical markers of bone metabolism, fracture rate, and clinical evaluation. RESULTS: Areal BMD and aBMD z scores showed significant improvement during the treatment period. Both serum and bone-specific alkaline phosphatase values were significantly decreased. Fracture rate reduced significantly from 3.5 +/- 1.01 to 0.83 +/- 0.77 fractures/year. Bone pain, which was severe in five patients, disappeared just after the first cycle, and the activity and mobility of patients increased. aBMD and aBMD z scores were decreased 1.5 years after discontinuation, although not statistically significant. Annual fracture rate increased significantly. Bone pain recurred in four patients. Pamidronate treatment was reinstituted in five of these patients at the end of 1.5 years. CONCLUSION: Cyclical pamidronate treatment is very effective in children with moderate/severe OI. This treatment should be started early enough before the occurrence of irreversible deformities and must be given for a longer time during the growth period. PMID- 18404975 TI - Neonatal screening for congenital adrenal hyperplasia: experience and results in Argentina. AB - AIM: To report the experience of the neonatal screening program for congenital adrenal hyperplasia (CAH) carried out in Buenos Aires, Argentina, from 1997 to 2006. POPULATION AND METHODS: 17-Hydroxyprogesterone (17OHP) was measured with an immunofluorometric assay in filter paper blood samples collected at neonatal maternity discharge. Filter paper blood levels <40 nmol/l were considered normal. 17OHP levels from 40-90 nmol/l triggered a new assessment to decide on a course of action. Confirmation of CAH was made with levels >90 nmol/l. This led to clinical follow up. For preterm (PT) infants, data were adjusted according to percentiles for gestational age and/or birth weight. RESULTS: From 80,436 screened newborns (46.8% girls), 8848 (11%) were PT. 15 term (T) and 3 PT infants were recalled (0.022%). Nine were confirmed as having CAH (8 T and 1 PT) (female/male: 0.8; incidence 1:8937). Mean ages of screening and treatment were 5.7 and 13 days. Only 33% of affected children were clinically suspected of having CAH prior to screening. Four boys and two girls presented salt-wasting forms and severe adrenal insufficiency crises were prevented as a result of the screening. CONCLUSIONS: Our findings confirm the benefits of CAH neonatal screening in our country with a high incidence of the classical form. Established criteria of screening and follow up allowed us to detect unrecognized affected males and females and to successfully prevent salt-wasting crises. PMID- 18404976 TI - Determination of iodine nutrition and community knowledge regarding iodine deficiency disorders in selected tribal blocks of Orissa, India. AB - AIM: To determine the status of iodine nutrition and knowledge of iodine deficiency disorders (IDD) in selected tribal mountainous blocks of Orissa, India. DESIGN: A community-based survey was performed, adopting the 30-cluster sampling and surveillance methodology for assessment of IDD recommended by WHO/UNICEF/ICCIDD. SUBJECTS: School-age children (6-12 years) and their mothers. METHODS: Total goitre rate (n=623) and urinary iodine excretion of children (n=530), iodine content in edible salt (n=505) and water (n=21) were measured. Community knowledge regarding IDD and awareness of iodized salt (n 20) was assessed. RESULTS: Total goitre rate was 23.6%, of which visible goiter was 6.9%. Prevalence of goiter increased with age in female and tribal children. Median urinary iodine was 38 Cmicro.Tl(-1) and 51.7% of children had urinary iodine values <100 pmicrogtl(-1) The mean iodine content for drinking water ranged from 1.22-3.6 pmicro.Tl(-1) Only 9.9% of salt samples had adequate iodine content (> or =5 ppm). Over 80% of respondents did not have knowledge of IDD and were not aware of salt iodization. CONCLUSIONS: Study results show moderate iodine deficiency with poor community knowledge of iodine nutrition. There is need to strengthen the monitoring of salt iodization and intensive education activities in the tribal areas. PMID- 18404977 TI - Pediatric hypothyroidism presenting with a polymyositis-like syndrome and increased creatinine: report of three cases. AB - We report three pediatric cases of primary hypothyroidism presenting with musculoskeletal complaints, elevated muscle enzymes and increased creatinine. Thyroid hormone replacement led to improvement in both the clinical features and laboratory abnormalities. PMID- 18404978 TI - Hypokalemic rhabdomyolysis in a child with 11-hydroxylase deficiency. AB - Myopathy is a well-recognized complication of persistent hypokalemia. Although hypokalemic myopathy may be due to divers diseases or to drug administration, hypokalemic rhabdomyolysis as a complication of under-treated 11-hydroxylase deficiency has not previously been described in the literature. We describe a 10 year-old boy with under-treated 11-hydroxylase deficiency who developed rhabdomyolysis following severe hypokalemia. Patients with under-treated 11 hydroxylase deficiency may present with hypokalemia in association with muscle weakness; if serum potassium is markedly low, rhabdomyolysis may occur. Hypokalemia-induced rhabdomyolysis should be carefully followed. PMID- 18404979 TI - Re: "Prevalence and clinical features of type 1.5 diabetes in children", Purushothaman R et al., JPEM 2007; 20: 981-987. PMID- 18404980 TI - Re: "Diabetes mellitus in Asian Indian children and adolescents", Nandkeoliar MK et al., JPEM 2007;20:1109-1114. PMID- 18404981 TI - Re: "Effect of metformin and rosiglitazone in a prepubertal boy with Alstrom syndrome", Sinha SK et al., JPEM 2007; 20: 1045-1052. PMID- 18404982 TI - [Expression of brain natriuretic peptide and c-fos gene in rat after acute myocardial ischemia and its medicolegal significance]. AB - OBJECTIVE: To study the mRNA expression of BNP and c-fos gene in rat heart after acute myocardial ischemia (AMI) and to provide a marker for its medicolegal diagnosis. METHODS: AMI animal model of rat was made by ligating LAD. mRNA expression of BNP and c-fos gene were studied with RT-qPCR and ordinary PCR at 10 min, 30 min, 60 min and 3h after the successful ligation. The H&E staining was also used. Changes of the mRNA expression in different time groups were compared. RESULTS: There was significant difference in BNP mRNA expression of the 3 h group by RT-qPCR compared with normal control group, 10 min, 30 min, and 60 min groups (P < 0.05). There were dramatic differences in c-fos mRNA expression between every two groups (P < 0.05) except between the normal group and the 10 min group, between the 30 min group and the 3 h group. The peak of c-fos expression was in 60 min group. No difference was shown between groups by the ordinary PCR. Myocardial fiber acidophilia staining and wavy changes could be seen occasionally at 3 h experimental group by H&E staining. CONCLUSIONS: C-fos gene probably be used as an auxiliary test for myocardial ischemia of duration of 30 minutes or longer. RT-qPCR may be suitable for diagnosis of early AMI. PMID- 18404983 TI - [Time-dependent expression of caspase-3 during contused skeletal muscle wound healing in rats]. AB - OBJECTIVE: To investigate the expression of caspase-3 during contused skeletal muscle wound healing in rats and to explore the applicability of caspase-3 in wound age estimation. METHODS: Male Sprague-Dawley rats undergone anesthesia with 2% pentobarbital sodium (45 mg/kg) were struck by a single strike at the skeletal muscle of the right leg, then were sacrificed by cervical dislocation at different post-traumatic intervals. The changes of caspase-3 activity were assessed by imunohistochemistry, Western blot and caspase-3 colorimetric assay. The skeletal muscle of non-contusion rat's were used as controls. RESULTS: No caspase-3 was detected in skeletal muscle in the control group. The expression level of caspase-3 in the peripheral area of contused rat skeletal muscle increased gradually and maximized on the fifth day after injury, and then decreased gradually. Caspase-3 was also detected in the regenerated skeletal muscle within 7-14 days after injury. CONCLUSION: The expression of capase-3 in the peripheral area of contused rat skeletal muscle indicated that it may play a pivotal role in apoptosis of the wounded skeletal muscle and may be used as a marker for wound age estimation. The expression of capase-3 in regenerating skeletal muscle suggests that caspase-3 activity may be used for skeletal muscle differentiation. PMID- 18404984 TI - [Coma criterion and classification standard of mild and moderate traumatic brain injury in rats]. AB - OBJECTIVE: To set up a classification standard of mild and moderate traumatic brain injury, for the purpose of reliable data comparison derived from different laboratories. METHODS: Traumatic brain injury (TBI) in rats was prepared by using a metallic pendulum-striker device. After injury, five variable parameters including the time of apnea and the areflexia, time of corneal reflex, external auditory canal stung reaction, body-righting reflex and needling reaction were determined and scored by using rat coma criterion. These data were judged and classified into mild and moderate head injury by brain patho-anatomy changes. Then the data were used to set up a multivariate discriminate equation. RESULTS: The distinguished probability of mild and moderate TBI according to actual direct measured value and the criterion were 88.9% and 91.9%, respectively. CONCLUSION: This method is able to classify mild and moderate TBI in rats. PMID- 18404985 TI - [Correlation between postmortem intervals and the changes of electrical conductivity in vitreous humor in rabbits after death]. AB - OBJECTIVE: To investigate the correlation between postmortem interval (PMI) and the changes of electrical conductivity in vitreous humor in rabbits after death. METHODS: The changes of electrical conductivity in vitreous humor in rabbits were measured using the conductivity meter under 30 degrees C during 0-48 hours and 20 degrees C during 0-120 hours after death. RESULTS: Electrical conductivity in vitreous humor in rabbits increased gradually under 30 degrees C and 20 degrees C from 0 to 48 hours and from 0-120 hours after death. The formulae of the relationship between PMI and conductivity under 30 degrees C and 20 degrees C were obtained by statistical analysis and the correlation coefficients were 0.970 and 0.983 (both P < 0.01), respectively. CONCLUSION: The increase of electrical conductivity in vitreous humor in rabbits after death may be used as the relatively objective parameter for PMI estimation. PMID- 18404986 TI - [Development of hand-wrist bones of 14 year-old adolescents. II. Standard of bony age for girls]. AB - OBJECTIVE: To study the current development of hand-wrist bones of adolescents in China, and establish special forensic bony age standard for estimating 14 years of criminal responsible age in female adolescents. METHODS: One hundred and ten healthy girls aged from 13 years and 9 months to 14 years and 3 months in Lengshuijiang district of Hunan province were enrolled in this study. X-ray films of their left hand-wrists were taken dorsaventrally, and 15 sites on the wrist x ray films were examined and measured under standard condition. The development grade was confirmed by maximal percentage to establish a method to estimate the bony age, which was then checked by single blind method. RESULTS: Development grade of the radius, the middle third phalanx and the fourth phalanx was partially closed (> 2/3), and the rest was wholly-closed with existence of the epiphyseal line. The development grade of the middle third phalanx and fourth phalanx was different between the younger group (< 14 years, partially closed, > 2/3) and the older group (> 14 years, mostly closed with existence of only epiphyseal line). The cross diameter of the osteoepiphysises were wider than that of all the diaphysises. The confirmed accuracy (in 30 girls) was 83.3%. CONCLUSION: Our study seemes to be feasible in establishing the estimation standard for bony age determination. The standard established by developmental metaphysis and authropometry may be particularly helpful in forensic practice for estimation of accurate criminal responsible age in adolescents. PMID- 18404987 TI - [Staging methods of skeletal growth by X-ray in teenagers]. AB - OBJECTIVE: To establish staging criteria for teenager's skeletal growth by examining osteoarticular X-rays. METHODS: Review publications on skeletal growth staging systems by domestic and overseas authorities. Abided by the staging principles on theory of cartilage epiphyseal growth and X-ray images of skeletal growth, X-ray imaging characteristics were studied through three hundred male and female teenagers epiphyseal growth of both sternal ends of clavicle and the left side of shoulder, elbow, carpal, hip, knee and ankle joints transversally and longitudinally. The subjects were from 11 to 20 years old, selected from Henan, Hainan and Zhejiang province in China. Twenty-four skeletal growth indexes were selected closely related to age. RESULTS: Based on regular changing characteristics of epiphyseal growth at different joints, 24 skeletal development indexes were divided into 2 to 8 stages. The stage number of different indexes of the single joint were similar. It was shown that the time of different epiphyseal growth in identical joints had isochromism. CONCLUSION: These staging criteria have merits of extensive skeletal growth indexes, and are applicable to wide age range with a strong maneuverability. It provides a favorable fundamental platform for forensic evaluating skeletal age in living subjects in China. PMID- 18404988 TI - [Stature estimation from the cervical vertebrae of living male by measuring Xray films of computer radiography]. AB - OBJECTIVE: To establish the regression equations and to evaluate the feasibility of measuring dimensions of cervical vertebrae with lateral films of computer radiography (CR). METHODS: Anterior heights and posterior heights of C3-C7 were measured in the lateral films of the cervical vertebrae in 150 adult males from Sichuan Han population in China The body height of each individual was recorded. Linear regression analysis between the height of the cervical vertebrae and body height was performed to establish the regression equations for stature estimation. In addition, the paired t-test between the predictive value and the actual body height, the residual analysis, the test of equal scedasticity and colinearity were performed in all equations. RESULTS: Thirty one statistically significant simple regression equations and 20 statistically significant multiple regressions equations were established. The coefficiencies of multiple correlations were from 0.656 to 0.259 and the standard errors of estimate were between 4.30 cm and 5.79 cm. Conclusion All equations established could be used to estimate the body height of the Han population of Sichuan adult males. It could be helpful to estimate statures from the CR films based on the dimensions of cervical vertebrae. PMID- 18404989 TI - [Detection of MMP-11 from menstrual blood using immunohistochemistry]. AB - OBJECTIVE: To prove the feasibility of detecting menstrual blood as well as its cellular localization with rabbit-anti-human matrix metalloproteinase-11 (MMP-11) polyclonal antibody. METHODS: MMP-11 in menstrual blood, peripheral blood, vaginal liquid, aged menstrual bloodstain, and endometrium sections were assayed with SAP immunohistochemistry. RESULTS: MMP-11 was found only in menstrual samples within stroma and epithelium cells. CONCLUSION: MMP-11 polyclonal antibody may be applied in the distinction between menstrual blood and venous blood. PMID- 18404990 TI - [STR genotyping from trace epithelial cells on fountain pen]. AB - OBJECTIVE: To evaluate the feasibility of STR genotyping from trace epithelial cells on fountain pen and to discuss the impact of conservation time on DNA typing. METHODS: Seven fountain pens were separately used by each of the 17 volunteers 20 minutes per day for a month and then were preserved on day 1, 3, 5, 7, 14, 21, and 28. DNA was extracted from the epithelial cells on fountain pen by silicon bead and was genotyped by Identifier kit. The corresponding control samples were buccal swabs of the above volunteers. The detectable numbers of loci were counted for assessment. RESULTS: There were statistically significant differences in the DNA genotyping by detectable numbers of gene loci between buccal swabs and epithelial cells on fountain pen of different conservation times (P < 0.01). The differences of detectable numbers of loci between the epithelial cells on fountain pen preserved on day 1, 3, 5, 7, 14, 21, 28 and the corresponding oral swabs were also statistically significant (P < 0.01). More than 12 loci could be successfully genotyped in 41.2% samples from the epithelial cells on fountain pen if the tests were performed within 24 hours. CONCLUSION: The trace epithelial cells on fountain pen can be used as biological samples for personal identification, but the conservation time would have influence on the results of DNA genotyping. PMID- 18404992 TI - [Characteristics and mechanism of boat propeller injuries]. AB - OBJECTIVE: To summarize the characteristics and investigate the mechanisms of boat propeller injuries so as to explore the identification methods between boat propeller injuries and corpse dismemberment. METHODS: More than 100 autopsy cases of boat propeller injuries were collected in a period between 1994 and 2005 in Huzhou district, Zhejiang province. The characteristics of injuries caused by propeller, including abrasion, wound, fracture and severed wound, and the characteristics of clothing, were retrospectively studied and summarized. The severed cross wound section of boat propeller injuries was compared with that caused by corpse dismemberment. RESULTS: The boat propeller injuries were resulted from high-speed propellers with enormous splitting power and mechanical cutting, while corpse dismemberment were resulted from cutting and dismembering the body with sharp instruments. Due to the different mechanisms, the different strength of force and recoil force, the severed wound cross section had different characteristics. CONCLUSION: Wounds caused by boat propeller injuries have their unique characteristics, distinguished from wounds of dismembered corpse. PMID- 18404991 TI - [Determination of ketamine and norketamine in blood and urine by high performance liquid chromatography]. AB - OBJECTIVE: To develop a high-performance liquid chromatographic method for determination of ketamine and norketamine in blood and urine. METHODS: The compounds were extracted from blood or urine by liquid-liquid extraction using toluene after blood or urine was adjusted pH to 14. The extracts were analyzed by HPLC. RESULTS: Linear limits of ketamine and norketamine determination in blood ranged from 0.05 microg/mL to 10 microg/mL (R2 > 0.9993) and in urine ranged from 0.01 microg/mL to 200 microg/mL (R2 > 0.9995). Limits of detection (LODs) for ketamine and norketamine were 0.006 microg/mL and 0.003 microg/mL (S/N > or = 3), respectively. The mean extraction recovery was over 82.4% and its coefficients of variation were less than 10.0% for ketamine and norketamine. Concentration-time curves and urinary drug velocity curves of ketamine norketamine were obtained by determinations of them in blood and urine in rat using the developed method. CONCLUSION: The method is sensitive, simple, rapid and suitable for determination of ketamine and norketamine in blood and urine for toxicological and clinical pharmaceutical analysis. PMID- 18404993 TI - [How to validate a bio-analytical method]. AB - It is imperative that any newly established bio-analytical method is validated thoroughly, using standardised parameters. The purpose of this article is to provide recommendations on how to validate a new bio-analytical method. Based on author's personal experience and some interesting discussion points from the conference of "International Association of Forensic Toxicologists" in 2007, the authors propose these essential requirements for validating a new analytical method. The key parameters of method validation include selectivity, linearity, accuracy, precision, LOD (limit of detection), LLOQ (the lower limit of quantitation), stability and the extraction recovery. For any bio-analytical method using LC-MSn (Liquid chromatography-mass spectrometry), studies of matrix effect should also be included in addition of the above parameters. PMID- 18404994 TI - [The recent development on age estimation]. AB - The main methods of age estimation at present mostly rely on characteristic physical and chemical changes of skeleton, soft tissue, cells, biomacromolecules and the other substances. Because physiological changes of tissues and organs during growth are extremely complicated, all methods have their limitations. In practical, it is necessary to combine several methods together in order to improve the accuracy of age estimation. This article reviewed recent development on age estimation domestically and abroad and discussed advantage and disadvantage of different methods. PMID- 18404995 TI - [Advancement in the study on electrophysiology of color vision and its applicability in forensic medicine]. AB - Color vision is a major component of vision function. Examination of color vision mostly relies on subjectivity at present. It is important to establish an objective method of color vision examination that would have particular clinical significance and forensic medicine value. With the development of new technology, the examination of visual electrophysiology has become a basic method of clinical diagnosis and investigation. Electroretinogram (ERG) could discriminate different cone cells and color abnormality. Visual evoked potential (VEP) generated characteristic negative wave after stimulation with different colors. Different visual electrophysiology detection methods were reviewed and in particular important new advancement of color vision research of ERG, F-VEP and P-VEP were introduced. The application prospect of color vision research in the clinical forensic medicine was also discussed. It is likely that color vision research study and visual electrophysiology will provide objective evaluation of retina and brain function. PMID- 18404996 TI - [Endometriosis: from research to clinical practice]. AB - Endometriosis, defined by the development of endometrial tissue outside the uterus, is a benign disease responsible for infertility and pelvic pain. The diagnosis based on a detailed gynaecological history and a careful clinical examination should be done as early as possible in order to treat patients correctly. Medical treatment is not appropriate in all cases and surgical treatment should be proposed but morbidity is related to the severity of the lesion. Ectopic implantation of endometrial cells needs complex interactions between host tissue and epithelial endometrial cells. The conditions for the development of endometriosis are oestrogeno-dependent growth of endometrial cells, induction of angiogenesis and lymphangiogenesis. Principal cellular and molecular factors of angiogenesis, lymphangiogenesis and fibrosis should be identified in order to develop new therapeutic strategies of endometriosis. PMID- 18404998 TI - [Hypogonadism due to LH deficiency]. AB - A 30-year-old man was investigated for delayed puberty and infertility. These investigations showed a complete absence of circulating luteinizing hormone (LH). Genetic studies revealed a missense mutation in the LHbeta gene (G36D). This mutation disrupts a vital cystine knot motif and abrogates the heterodimerization and secretion of LH. Treatment with hCG was instituted, which led to arise in testosterone and improvement in spermatogenesis. After in vitro fertilization the patient had a son who was heterozygous for the G36D mutation. A second patient with similar clinical and biological presentation has been explored. A non-frame shift deletion of 3 base-pairs was discovered at position 20 which led to the deletion of a lysine residue. The proband and his prepubescent brother were homozygotic for this mutation. These cases illustrate the important physiological role of LH in male sexual maturation and fertility. PMID- 18404997 TI - [Role of interleukin-9 in asthma and allergic reactions]. AB - Interleukin 9 (IL-9) is a cytokine produced by activated T lymphocytes and that activates in vitro mast cells as well as T and B lymphocytes. In vivo, transgenic mice overexpressing the gene encoding IL-9 show several of the hallmarks of human allergic asthma: increased IgE concentration, bronchial mastocytosis, eosinophilia, increased mucus production, as well as bronchial hyperresponsiveness. Whereas some of these features reflect direct IL-9 activities on target cells such as mast cells and B lymphocytes, increased mucus production and eosinophilia rather result from IL-13 and IL-5 production induced by IL-9 in T lymphocytes and mast cells. Preclinical studies in mice have shown that anti-IL-9 blocking antibodies interfere with the development of asthma-like reactions. In the human species, asthmatic patients produce large amounts of this cytokine and IL-9 production correlates nicely with species biological parameters of the disease. Phase 2 clinical trials are in progress to test the efficacy of anti-IL-9 antibodies in humans. PMID- 18404999 TI - [Multiple post-translational modifications implied in the gene expression regulation: example of the sumoylation of Ets factors]. AB - To regulate the spatiotemporal expression of their target genes, the transcription factors undergo post-translational modifications of which the most studied is phosphorylation. Acetylation and ubiquitinylation on lysine residues also exert a role in the transcription, as it is the case for the regulation of the activity of the huge family of Ets transcription factors. Recently, sumoylation, a post-translational modification similar to ubiquitinylation, was described as playing a crucial role in the inhibition of the activity of these factors. PMID- 18405000 TI - [p27Kip1 independently promotes neuronal differentiation and migration in the cerebral cortex]. AB - The generation of glutamatergic neurons by stem and progenitor cells is a complex process involving the tight coordination of multiple cellular activities, including cell cycle exit, initiation of neuronal differentiation and cell migration. The mechanisms that integrate these different events into a coherent program are not well understood. Here we show that the cyclin-dependent kinase inhibitor p27Kip1 plays an important role in neurogenesis in the mouse cerebral cortex, by promoting the differentiation and radial migration of cortical projection neurons. Importantly, p27Kip1 promotes neuronal differentiation and neuronal migration via two distinct mechanisms, which are themselves independent of the cell cycle regulatory function of p27Kip1. p27Kip1 inactivation by gene targeting or RNA interference results in neuronal differentiation and radial migration defects, demonstrating that p27Kip1 regulates cell migration in vivo. The differentiation defect, but not the migration defect, is rescued by overexpression of the proneural gene Neurogenin 2. p27Kip1 acts by stabilizing Neurogenin 2 protein, an activity carried by the N-terminal half of the protein. The migration defect resulting from p27Kp1 inactivation is rescued by blocking RhoA signalling, an activity that resides in the c-terminal half of p27Kip1. Thus, p27Kip1 plays a key role in cortical development, acting as a modular protein that independently regulates and couples multiple cellular pathways contributing to neurogenesis. PMID- 18405001 TI - [Molecular and cellular characterization ion of IKAP protein and the Elongator complex. Implications for familial dysautonomia]. AB - Familial dysautonomia (FD), a severe neuro-developmental and neurodegenerative genetic disorder, is caused by mutations of IKBKAP encoding a subunit of Elongator. FD patients have decreased expression of IKAP in a tissue-specific manner and consequently impaired Elongator function. The biological roles of human IKAP/Elongator remained elusive for a while. However, recent data based on the generation of cellular loss of function models of IKAP through RNA interference strongly suggest a role for this protein in transcriptional elongation. Other data also provide evidence for a role of Elongator in tRNA modifications. Importantly, cells depleted for IKAP have defects in cell motility because of impaired transcriptional elongation of some genes coding for proteins involved in cell migration. Therefore, cell motility deficiency seen in IKAP depleted cells may underlie the neuropathology of FD patients. PMID- 18405002 TI - [Development of new methods of magnetic resonance for the characterization of oxygenation and perfusion of tumors. Diagnostic and therapeutic applications]. AB - Tumor hypoxia is a key factor of the tumor growth and response to therapy. In this short note, we describe our recent contribution to the development of new non-invasive methods for measuring tumor oxygenation and perfusion by magnetic resonance techniques (NMR and EPR). After validation, these techniques have been applied to study the influence of physiological and pharmacological factors on tumor hemodynamics. The non-invasive study of the changes in the tumor microenvironment allowed the elaboration of rational schedules to potentiate the anti-cancer therapy when combining treatments. PMID- 18405004 TI - [Medical certificates: law, deontology and practice]. AB - At the start of the 21st century, in a society that gives the impression of switching century every five years because of its never-ending evolution, the general practitioner is confronted every day to more complex situations, including problems encountered at the legal level. These last few years, the legal interrogations relevant to medicine and its progress became considerably more extensive in Belgium, as in many other countries. The medical law and its regulation are the subject of many front titles in the press and of animated political debates. The main issues concern the organ transplantation, the meddling with human genetics, the questions related to the reproduction, euthanasia or the protection of the patient's rights. All these legal problems, without exception, are the subject of a great deal of attention in society and create to many controversies. The growing number of regulations in many fields of medicine, as well as their growing complexity, reinforces the importance of the medical law as a distinct discipline of legal sciences. More and more, the doctors of any qualification are required to guarantee, by adequate certificates, the health, the aptitude, the situation of disease of their patients. The patients can then have certain advantages granted by society. Out of concern for their patients, the experts are sometimes tempted not to be perfectly objective. This attitude can lead to penal or disciplinary sanctions and may undermine the confidence of people who exercise authority to give the requested advantages. The initial mistrust, which led to the system of the certificate, extends to those who were supposed to fully collaborate. How to leave this infernal round that sterilizes, denatures the medical act? While waiting for men of goodwill to consider the system and reform it, it remains with the writers of certificates to avoid the traps lying in their path. The purpose of this work is to propose models of certificate that experts can fill in accordance of the circumstances. It is only a guide and consequently, it sometimes requires an interpretation. This is why each model is described together with its justification and its traps. PMID- 18405003 TI - [Glioblastomas are resistant to apoptosis but less resistant to the autophagic process]. AB - Malignant gliomas of which glioblastomas represent the ultimate grade of malignancy are characterized by dismal prognoses because malignant glioma cells present both important proliferation and neoangiogenesis processes and can actively migrate through the narrow extra-cellular spaces in the brain, often travelling relatively long distances, making them elusive targets for effective surgical management. Invasive malignant glioma cells show a decrease in their proliferation rates and a relative resistance to apoptosis (type I programmed cell death) but seem less resistant to autophagic cell death (type II programmed cell death). Autophagic cell death represents an alternative mechanism to overcome, at least partly, the dramatic resistance of glioblastomas to pro apoptotic-related therapies. Another way to potentially overcome apoptosis resistance is to decrease the migration of malignant glioma cells in the brain, which then should restore a level of sensitivity to pro-apoptotic drugs. We conclude this work with an algorithm showing the optimal current treatment for glioblastoma with the potent future innovations. PMID- 18405005 TI - [Discovery of the splicing of peptides by the proteasome and study of preclinical models of anticancer immunotherapy]. AB - Our scientific activity belongs to tumor immunology: in the last fifteen years, we have contributed to the molecular definition of tumor antigens recognized by cytolytic T lymphocytes, both in human and mice, and to the validation of cancer immunotherapy in appropriate preclinical models. Here we describe two findings that we recently made in this field. The first is the discovery of a new mechanism of production of antigenic peptides, based on the splicing of peptides by the proteasome. The second is the discovery of an important mechanism of tumor resistance to immune rejection. The major interest of this mechanism is that it can be fought with pharmacological agents, which could be used to boost the efficacy of immunotherapy in cancer patients. PMID- 18405006 TI - Gene therapy for haemophilia "A" and "B": efficacy, safety and immune consequences. AB - The first successful gene therapy trials for the treatment of hereditary disorders underscore the potential of gene therapy to combat disease and alleviate human suffering. The development of gene therapy for haemophilia is not only a research priority in its own right but also serves as an ideal trailblazer for many different diseases. Significant progress has recently been made in the development of gene therapy for the treatment of haemophilia A and B. Long-term therapeutic levels of factor VIII and IX could be expressed following gene therapy in haemophilic mice, stably correcting the bleeding diathesis. These advances parallel the development of improved gene delivery systems. The induction of neutralizing antibodies (inhibitors) to the clotting factors could potentially preclude stable phenotypic correction. The risk of inhibitor formation varied, depending at least in part on the type of vector used and its in vivo tropism. We also demonstrated that the risk of immune responses to the vector particles, the clotting factors and/or transduced cells can be reduced by using vectors that only minimally interact with antigen presenting cells. In haemophilic mice, robust and stable clotting factor expression levels were achieved using adeno-associated viral vectors based on the newly disovered serotypes AAV8 and AAV9 which can efficient deliver the clotting factor genes into hepatocytes without triggering any inflammatory responses or adverse events. Pre-clinical studies in large animal models will be initiated to further validate these improved AAV vectors to ultimately justify a clinical trial in patients with severe haemophilia. PMID- 18405007 TI - Professional education a priority for nursing's first health minister. Interview by Tamsin Snow. PMID- 18405009 TI - What must be done. AB - Has nursing lost its way? Reports highlighting poor standards of care have prompted a period of reflection. Our series has examined the issues and suggested the way forward. PMID- 18405010 TI - School of thought. AB - Kidbrooke School in south east London faced several health challenges among its students. A service run by a school nurse has set about addressing them. PMID- 18405011 TI - Building lives. AB - Our series on expertise in nursing looks this week at a clinical nurse specialist in bipolar disorder. PMID- 18405012 TI - Nurses have no business killing. PMID- 18405013 TI - Lessons in sex. PMID- 18405014 TI - Solutions, techniques and pressure in wound cleansing. AB - This information on best practice discusses the evidence on the use of solutions, techniques and pressure in wound cleansing. The article has been reproduced with the permission of the Joanna Briggs Institute (JBI). The article (JBI 2006a), which updates and supersedes the 2003 information sheet of the same name, has been derived from a systematic review conducted in 2004 (Fernandez et al 2004). The primary references on which this information sheet is based are available in the systematic review reports to members of the JBI via the website: www.joannabriggs.edu.au PMID- 18405015 TI - Enterococcal infection. AB - This article describes enterococci bacteria and the measures related to glycopeptide-resistant enterococci, including screening, treatment and management, antibiotic prescribing and infection control. PMID- 18405016 TI - Urinary catheterisation. AB - This article examines the benefits and risks associated with urinary catheterisation and how these risks can be reduced. PMID- 18405017 TI - Ocular complications associated with diabetes mellitus. AB - Diabetes is a progressive disease that may adversely affect healthy functioning of the body. The eyes are often indicators of systemic disease and are particularly vulnerable to the pathophysiological changes that occur as a result of diabetes. This article provides an overview of five main ocular complications associated with diabetes, cataract, diabetic retinopathy, central retinal vein occlusion, central retinal artery occlusion and neovascular glaucoma. The assessment and treatment of such complications and the role of the nurse in the care and management of patients are discussed. PMID- 18405018 TI - Female urinary catheterisation. PMID- 18405019 TI - How to take the flak. PMID- 18405020 TI - Remote solutions. PMID- 18405021 TI - The balance of power. Interview by Hazel Walker. PMID- 18405022 TI - [Acquired male urethra diverticula. Report of seven cases. Bibliographic review]. AB - OBJECTIVES: To report our personal experience in a rare urethral pathology in relation with iatrogenic sequelae from surgical treatment of lower urinary tract pathologies. METHODS: We analyze the causes, type of presentation, and surgical treatment of seven cases of urethral diverticula, which account for 2.18% of the urethral procedures performed in our department, this latter representing 2.3% of a total of 11,845 surgical procedures in a period of 25 years. RESULTS/CONCLUSIONS: Male urethra diverticula go unnoticed because their initial symptoms are very similar to other lower urinary tract entities. But there is a definitive sign once the case is advanced: the appearance of a "lump" in the ventral side of the anterior urethra, the compression of which empties urinary content, sometimes fetid; or also the feeling of having a bag with stones if they host lithiasis. Surgical reconstructive treatment is very important to guarantee the absence of an obstacle distal to the cavity as well as to achieve a consistent ventral floor for the urethra to avoid recurrence, and that, as we will see, maybe obtained by various procedures. The attention to the original disease which indirectly had been the cause of the problem completes the therapeutic scheme for each case. PMID- 18405023 TI - [Perinephric abscess: associated factors and mortality]. AB - OBJECTIVES: To determine predisposing factors in perinephric abscesses and to find events associated with unfavourable outcome. METHODS: We carried out a clinical, descriptive, retrospective and cross-sectional study, including 23 patients diagnosed of perinephric abscess admitted to our hospital. RESULTS: In patients with perinephric abscess, clinical charts included diabetes mellitus in 65.2%, history of nephrolithiasis in 43.47% and history of urological surgery in 17.38%. On hospital admission, haemoglobin greater than 10.5 g/dL and white blood cell count lower than 15 x 10(3) / microL were associated with nephrectomy, and platelet count lower than 140 x 10(3) / microL with septic shock. General mortality was 8.69%, and 78.3% if patient required nephrectomy. Patients who died had fever, anaemia, white blood cell count greater than 16 x 10(3) / microL, platelet count lower than 130 x 10(3) / microL, and hyponatremia of 125 mEq/L or lower at hospital admittance, and all of them had septic shock and required nephrectomy. CONCLUSIONS: In our series, a higher percentage of patients had diabetes mellitus and/or nephrolithiasis. Haemoglobin level and white blood cells count were associated with loss of the renal unit, thrombocytopenia was associated with septic shock and hyponatremia with mortality. PMID- 18405024 TI - [Transurethral resection in prostate cancer, a radical procedure. Experience with 1017 cases]. AB - OBJECTIVES: From the beginning of TUR in 1931 the reseccionists thought of resecting prostate cancer. Execution however failed for deficient instruments and techniques. The first transurethral resection for prostate cancer: TURPC--was performed at our institution in 1957 by Hans J. Reuter. Low pressure irrigation enabled safe transurethral resection of the prostate including the capsule. Thus we started in 1985 a prospective study to verify TURPC as a radical procedure. METHOD: TURPC requires continuous low pressure irrigation with irrigating reservoir fixed at the operating table. The liquid level within is to maintain less than 20 cm. water above the pubic region in lithotomy position. Continuous flow is maintained by suprapubic trocar and a resectoscope with 28Fr. sheath. An autoregulated electro-surgical unit is indispensable. It automatically adjusts the high-frequency current to suit the cut tissue's electrical resistance for precise cutting. Videomonitoring is mandatory. The prostate is resected completely with its capsule into periprostatic fat together with the seminal vesicles. The specimen is retrieved in fractions to guarantee correct histopathological staging. If indicated laparoscopic staging lymphadenectomy is performed. A secondary session for control of positive margins follows after 12 weeks. RESULTS: From 1985-2004 1,017 patients with a mean age of 68.9 years and with clinically localized prostate cancer were resected by 5 surgeons with curative intention. The cancer stage distribution was in %: T1: 12, T2: 43, T3: 41, T4: 4.--G1: 8, G2: 72, G3: 20. Mortality 1 out of 1,017. Bladder neck incision 8.9%. Incontinence grade 3: none. Postoperative survival over 10 years was 82% in T1, 87% in T2, 81% in T3 and 34% in T4 patients. Biochemical recurrence as of the rise of three consecutive PSA-values was within 5 years 15% in T1, 19% in T2, 27% in T3. CONCLUSIONS: Considering our results we conclude that prostate cancer can be resected transurethrally as radical as with open surgery. The outcome as to survival and PSA-recurrence is comparable, the incontinence rate is less then with open surgery. It is mandatory to have low pressure irrigation with suprapubic trocar, 28Fr. sheath and on autoregulated electrosurgical unit, video monitoring and a well experienced reseccionist. PMID- 18405025 TI - [Current status of the diagnosis and treatment of Leydig cell testicular tumor. Report of two cases]. AB - OBJECTIVES: To review the current diagnosis and treatment of Leydig cell testicular tumors, with special attention to conservative treatment. METHODS: We report two cases of Leydig cell tumor in young adults, diagnosed two years after the appearance of bilateral gynecomastia as first clinical symptom. RESULTS: One year and eight months after orchyectomy, respectively, clinical symptoms completely disappear in the first case and significantly improved in the second. CONCLUSIONS: Around 10% of the cases, and only in adult patients, these tumors may be malignant, being radical orchyectomy the treatment of choice, although conservative surgery may be performed. Independently of the operation, follow-up must be prolonged. PMID- 18405026 TI - [Urinary lithiasis secondary to medication in HIV+ patients receiving Indinavir]. AB - OBJECTIVES: Therapy with protease inhibitors is commonly used in patients infected by human inmunodeficency virus (HIV). 20% of the administered dose is excreted by the kidney, and when alkaline urine is present, indinavir may crystallize forming stones and patients may experience renal colic due to this fact. METHODS: Between January 1998 and June 2005, 26 patients receiving antiretroviral treatment with protease inhibitors received care at our hospital because of renal colic or flank pain. All of them underwent physical examination, echography and urography as well as blood and urine analysis. Patients were treated ambulatory excepting those in whom oral analgesics were insufficient to control the pain. RESULTS: All patients had been treated with indinavir for longer than 12 months. They represented 4% of all patients treated with the recommended dose of Crivixan. Most of them presented flank pain, associated in most cases to microhaematuria. Five of them required hospitalization because of persistent pain in spite of endovenous analgesia. Imaging tests (echography and urography) showed functional delay of the kidney (2 cases), ureteral stasis (4 cases) and little lithiasic concretions of mild radiologic density (5 cases). Urinalysis revealed suggestive christaluria and alkaline pH. All patients required hidratation and analgesic treatment. In 3 patients indinavir dose was reduced, it was retired in another one, and 100 mg of rito-navir were added in another one. Unsuccesfuly ureteral cateterization was tried in one patient. All of them presented symptomatic improvement. CONCLUSIONS: We ought to know the capability of indinavir to form urolithiasis in HIV patients treated with protease inhibitors, although its use is decreasing along time. Prevalence of urolithiasis in these patients seems to be higher as length of treatment becomes longer. Metabolic alterations in urine have been proved in these patients, contributing to a higher incidence of lithiasis than in general population. PMID- 18405027 TI - [Usefulness of cytokines as surgical aggression markers in the ischemia reperfusion syndrome and post transplant renal function in an experimental model of laparoscopic vs. open renal autotransplantation]. AB - OBJECTIVES: To analyze the modifications induced by laparoscopic and open nephrectomies in living donor transplantation on cytokines, to evaluate operative trauma of different surgical techniques and the influence on ischemia/reperfusion syndrome and renal function. METHODS: Thirty pigs underwent left nephrectomy, 15 by laparoscopy and 15 by open approach in an experimental autotransplantation model. RESULTS: Serum level of IL-2, IL-6, IL-10 and tumor necrosis factor (TNF) were lower during laparoscopic than open nephrectomy: 6.8 +/- 0.6 vs 13.9 +/- 1.1 pg/ml for IL-2, 46.2 +/- 2.3 vs 84.4 +/- 2.5 pg/ml for IL-6, 26.1 +/- 2.4 vs 92.8 +/- 12.6 pg/ml for IL-10, and 17.6 +/- 2.1 vs 38.5 +/- 4.8 pg/ml for TNF. There was no association between renal blood flow (RBF) and cytokines levels during nephrectomy: IL-2 (p = 0.498), IL-6 (p = 0.117), IL-10 (p = 0.081) y TNF (p = 0.644). However, there was correlation between IL-10 and the decrease of RBF after transplantation: (R2 0.48; p = 0.02). Initial serum creatinine levels were correlated with RBF and IL-2 levels during nephrectomy (R = 0.831, R2 = 0.691, p = 0.025), and postransplantation RBF (R = 0.784, R2 = 0.614, p < 0.0001). Seventh day creatinine levels were correlated with postransplantation RBF (R = 0.537, R2 = 0.289, p = 0.002) and IL-2 levels during nephrectomy (R = 0.685, R2 = 0.469, p = 0.015). CONCLUSIONS: Cytokine levels were higher during the open approach than laparoscopic procedure. High levels of RBF during nephrectomy and transplantation improve early graft function while low levels of RBF and high levels ol IL-2 during nephrectomy induce delayed graft function. PMID- 18405028 TI - [Hydraulic dilation (URETEROMAT): effective and safe ureteroscopy]. AB - OBJECTIVES: After the description and standardization of the technique of ureteroscopy in 1979, several methods to obtain a nontraumatic, simple, efficient and fast ureteral access have been developed. The hydraulic expansion of the ureter was possible in 1986, with the development of ureteromat. We evaluate the effectiveness of hydraulic ureteral expansion (Ureteromat) for ureteral access and the later endoscopic procedures. METHOD: The last 100 ureteroscopy carried out from February to October 2006 were evaluated. We used a semi-rigid 9.8 Fr. ureteroscope and hydraulic expansion (Ureteromat) was performed in the same way in all procedures since 1986. RESULTS: Of the total of cases, 92 were indicated for lithiasic pathology, 7 were exploratory and 1 for ureteral stricture. In 94 patients the procedure was carried out with hydraulic expansion alone, in 2 cases we used a balloon catheter, in 3 patients expansion of the meatus was done with forceps and 1 procedure was not possible due to an ureteral stop. CONCLUSION: The Ureteromat pump guarantees an excellent hydraulic expansion of the ureteral meatus and the passage of the intramural ureter. The visibility is very good at any moment of the procedure, making ureteroscopy a simple, effective, non traumatic and fast method. PMID- 18405029 TI - [Inflammatory myofibroblastic tumor. Case report]. AB - OBJECTIVE: To report one case of myofibroblastic bladder tumor. METHODS/RESULTS: We report the case of a 30-year-old woman with recurrent urinary tract infections and incidental finding of a bladder mass after abdominal ultrasound. After TUR, inflammatory myofibroblastic tumor of the bladder was diagnosed. CONCLUSION: Myofibroblastic tumor (also known as inflammatory pseudotumor or pseudosarcoma) is a benign tumor with mesenchymal origin. Bladder location is very uncommon. It must not be misdiagnosed as a malignant neoplasm. Optimal treatment when arising in the bladder is TUR, with excellent long-term prognosis. PMID- 18405030 TI - [A new case of penile testicular ectopia]. AB - OBJECTIVE: To report a new case of penile testicular ectopia. METHODS: We present the case of a three-year old boy who was diagnosed of penile testicular ectopia and underwent surgery. RESULTS: Physical examination revealed on ovoid tumor in the center of the dorsal side of the penile root, together with empty left scrotum. Inguinal incision revealed the exit of the spermatic cord from the left superficial inguinal ring, which was directed to a testicle in a subdermal position in the root of the penis. Funiculolysis and orchyopexy were performed. CONCLUSIONS: The case here reported corresponds to a penile testicular ectopia of central localizations, with normal inguinal course of the spermatic cord. PMID- 18405031 TI - [Bellini collecting duct renal carcinoma: case report]. AB - OBJECTIVE: The collecting duct carcinoma (Bellini's carcinoma) is an unfrequent lesion; its macro and microscopic characteristics can help diagnosis. 65-year-old Colombian woman presenting left-flank and hypochondrio stub-type episodic pain associated with hematuria, nausea and weight loss. RESULTS: The diagnosis was of collecting duct carcinoma. CONCLUSIONS: The clinical characteristics, macrocospic features, histology and immunohistochemistry are analyzed; we also perform a bibliography review. PMID- 18405032 TI - [Thigh abscess: a very rare complication of retropubic adenomectomy]. AB - OBJECTIVE: Usual complications of retropubic adenomectomy are well-known. Despite their low incidence, urologists face them relatively often in their daily practice due to the frequent performance of this technique, and they usually have satisfactory outcomes. We want to attract attention to the presence of a thigh abscess as an exceptional complication of adenomectomy. It is such on oddity that may pose very important diagnostic problems, with negative implications due to treatment delay. The knowledge of it may be decisive at the time of diagnosis and enable a fast decision-making process. METHODS/RESULTS: We present the case ofa great thigh abscess as a complication of adenomectomy. We performed a detailed description of its presentation and avatars occurred until diagnosis and treatment, fortunately successful. We collect the pathogenic mechanisms of dissemination and comment on bibliography. CONCLUSIONS: The presence of a thigh abscess as a late complication of adenomectomy is an exceptional entity requiring a rapid diagnosis to an enable successful treatment, because it is a severe complication. PMID- 18405033 TI - [Supernumerary spleen simulating a renal tumor. Case report]. AB - OBJECTIVE: Report one case of ectopic spleen simulating a renal tumor. We performed a review of etiopathogenesis, diagnosis and treatment. METHODS: We present the case of a 54-year-old splenectomized female patient with the incidental diagnosis on ultrasound of a left kidney upper pole mass simulating a renal tumor. RESULTS: After the performance of CT scan, and with the suspicion of renal neoplasia, the patient underwent tumorectomy with the pathologic diagnosis of the specimen compatible with supernumerary spleen. CONCLUSIONS: We emphasize the importance of including pseudo masses of splenic origin in the differential diagnosis of left renal, adrenal and retroperitoneal tumors. We emphasize the importance of CT scan as the imaging test of choice, and gammagraphy as a complementary test. We point out conservative attitude as the most suitable in cases of asymptomatic pseudomass of splenic origin. PMID- 18405034 TI - [Urethral lithotripsy and cystolithotripsy of giant urethral stones through a transperineal approach]. AB - OBJECTIVE: To show that a one-stage transperineal procedure may be effective in the treatment of urethral stones secondary to urethral strictures. METHODS: We present the case of a 36 y/o paraplegic male with neurogenic bladder, history of intermittent catheterization, and urethral strictures treated with internal urethrotomies several times. Imaging tests showed the presence of giant urethral and bladder stones with a concomitant bulbar urethra stricture. A one-stage transperineal approach was indicated. RESULTS: Postoperative follow up was satisfactory. The patient was discharged from the hospital one day after the surgery. At 12 months there were no signs of urethral obstruction and he continued with intermittent catheterizations uneventfully. CONCLUSION: One stage procedure was indicated because previous endourological treatments were unsuccessful. To our knowledge there is no published case of urethral stones the size we report to or its treatment with lithotripsy and litholapaxy in a one stage transperineal approach. PMID- 18405035 TI - [Skin metastasis during follow-up of a clear cell renal carcinoma]. AB - OBJECTIVE: To report a new case of metastatic renal carcinoma with atypical localization. Skin metastatic involvement by urological tumors is a rare clinical entity. It is usually associated with internal organ involvement, so that this finding makes prognosis poor. We review the most frequent metastatic sites, their prognostic implications and treatment. METHODS: We present the case of a 60-year old male patient with the diagnosis of inferior maxillary skin metastasis 18 months after radical nephrectomy for clear cell adenocarcinoma. RESULTS: The patient was treated with oral Sorafenib within a phase II clinical trial. Two years after diagnosis the patient is alive and a disease-free. CONCLUSIONS: The wide variability of localizations and forms of presentation of metastatic renal carcinoma obliges to be alert during followup. New antiangiogenic drugs are an efficient therapeutic option. PMID- 18405036 TI - [Renal-duodenal fistula in the context of a renal tumor]. PMID- 18405037 TI - [Recurrent proximal migration of a double J ureteral catheter]. PMID- 18405038 TI - Bladder leiomyoma: case report. AB - OBJECTIVE: Leiomyoma is a benign lesion which represents 0.04-0.5% of bladder tumors. It is more common in females and its peak incidence is between 4th and 5th decades. Surgery is the treatment of choice and adequate results have been previously reported. METHODS: 38 years old male patient who consulted for chronic pelvic pain syndrome. CT scan showed a 2 cm diameter exophitic lesion at the anterior left lateral bladder wall, which protruded into the perivesical fat. We performed a laparoscopic partial cistectomy locating the tumor and resecting it with simultaneous cystoscopic control, obtaining negative margins. The operative time was 70 minutes with an intraoperative blood loss of 50 ml. Postoperative period was uneventful. Final pathology reported: Bladder wall leiomyoma, without mitosis or atypia. Immunohistochemistry was positive for Actine and Vimentine stablishing diagnosis. Cd 1 17 (c-kit) was negative and ruled out a Gastrointestinal Stromal Tumor. CONCLUSIONS: Leiomyoma is bladder's most common benign non epithelial tumor. It represents 35-46% of these lesions with a 2:5 male/female ratio. It origins from the smooth muscle bundles and at the urinary tract the most common localizations are kidney and bladder. Clinical presentation depends on tumor size and localization. Ultrasound is the most useful diagnostic tool and the pathological diagnosis is mandatory. Surgery is the treatment of choice and technique depends on tumor size and localization. The laparoscopic approach seems to be an effective alternative in this group of tumors. Prognosis is good and recurrence is rare. PMID- 18405039 TI - Segmental testicular infarction vs testicular tumour: the usefulness of the excisional frozen biopsy. AB - OBJECTIVE: To report a case of a Segmental testicular infarction (STI) simulating a testicular tumour and to discuss the importance of the excisional frozen biopsy. METHODS: We present the case of a patient with STI mimicking a testicular tumour. RESULTS: The patient was treated with partial orchiectomy after excisional frozen biopsy. CONCLUSIONS: The excisional frozen biopsy in testicular masses is a diagnostic maneuver to be considered in order to perform a testis sparing surgery. PMID- 18405040 TI - Keeping the "cognitive" in cognitive neuroscience, the "affective" in affective neuroscience, and the "behavioral" in behavioral neuroscience: the CABN mission for the next five years. PMID- 18405041 TI - The case for implicit category learning. AB - This article evaluates the evidence regarding the claim that people can learn a novel category implicitly--that is, by an implicit memory system that is qualitatively different from an explicit system. The evidence that is considered is based on the prototype extraction task, in which participants are first exposed to a set of category exemplars under incidental learning instructions and are then required to categorize novel test items. Knowlton and Squire (1993) first reported that memory-impaired patients performed normally on the prototype extraction task while being impaired on a comparable recognition task. Several studies have replicated these results, but other articles have criticized the evidence for implicit category learning on both methodological and theoretical grounds. In this article, we consider five of these criticisms-for example, that the normal performance of the patients is due to intact working memory mechanisms (see, e.g., Palmeri & Flannery, 1999) or to the lesser cognitive demands of prototype extraction rather than recognition (e.g., Nosofsky & Zaki, 1998). For each of the five criticisms, we offer counterevidence that supports implicit category learning. PMID- 18405042 TI - Dissociable neural subsystems underlie visual working memory for abstract categories and specific exemplars. AB - An ongoing debate concerns whether visual object representations are relatively abstract, relatively specific, both abstract and specific within a unified system, or abstract and specific in separate and dissociable neural subsystems. Most of the evidence for the dissociable subsystems theory has come from experiments that used familiar shapes, and the usage of familiar shapes has allowed for alternative explanations for the results. Thus, we examined abstract and specific visual working memory when the stimuli were novel objects viewed for the first and only time. When participants judged whether cues and probes belonged to the same abstract visual category, they performed more accurately when the probes were presented directly to the left hemisphere than when they were presented directly to the right hemisphere. In contrast, when participants judged whether or not cues and probes were the same specific visual exemplar, they performed more accurately when the probes were presented directly to the right hemisphere than when they were presented directly to the left hemisphere. For the first time, results from experiments using visual working memory tasks support the dissociable subsystems theory. PMID- 18405043 TI - Dynamic facial expressions of emotion induce representational momentum. AB - Two experiments were conducted to explore whether representational momentum (RM) emerges in the perception of dynamic facial expression and whether the velocity of change affects the size of the effect. Participants observed short morphing animations of facial expressions from neutral to one of the six basic emotions. Immediately afterward, they were asked to select the last images perceived. The results of the experiments revealed that the RM effect emerged for dynamic facial expressions of emotion: The last images of dynamic stimuli that an observer perceived were of a facial configuration showing stronger emotional intensity than the image actually presented. The more the velocity increased, the more the perceptual image of facial expression intensified. This perceptual enhancement suggests that dynamic information facilitates shape processing in facial expression, which leads to the efficient detection of other people's emotional changes from their faces. PMID- 18405044 TI - The medial temporal lobe and visual working memory: comparisons across tasks, delays, and visual similarity. AB - Whether the hippocampus and medial temporal lobe (MTL) play any important role in visual working memory is a relatively new and controversial research question. The primary goal of this study was to assess working memory for faces over very short delays in patients with MTL damage. Patients and matched controls were required to remember one face that was parametrically morphed to be more or less similar to a probe face, over either a 1- or an 8-sec delay. Memory was assessed using both forced choice and old-new recognition tasks. The results show that MTL damage impairs both speed and accuracy of visual working memory across tasks. We speculate that the hippocampus is generally necessary for memory encoding. PMID- 18405046 TI - I know how you feel: task-irrelevant facial expressions are spontaneously processed at a semantic level. AB - Previous studies have demonstrated that emotions are automatically processed. Even with subliminal presentations, subjects involuntarily mimic specific facial expressions, are influenced by the valence of a preceding emotion during judgments, and exhibit slowed responses to personally meaningful emotions; these effects are due to reflexive mimicry, unconscious carryover of valence, and attentional capture, respectively. However, perception-action effects indicate that rapid processing should involve deep, semantic-level representations of emotion (e.g., "fear"), even in the absence of a clinical emotion disorder. To test this hypothesis, we developed an emotional Stroop task (Emostroop) in which subjects responded nonverbally to emotion words superimposed over task-irrelevant images of faces displaying congruent or incongruent emotional expressions. Subjects reliably responded more slowly to incongruent than to congruent stimuli, and this interference was related to trait measures of emotionality. Rapid processing of facial emotions spontaneously activates semantic, content-rich representations at the level of the specific emotion. PMID- 18405045 TI - Affective priming of emotional pictures in parafoveal vision: left visual field advantage. AB - This study investigated whether stimulus affective content can be extracted from visual scenes when these appear in parafoveal locations of the visual field and are foveally masked, and whether there is lateralization involved. Parafoveal prime pleasant or unpleasant scenes were presented for 150 msec 2.5 degrees away from fixation and were followed by a foveal probe scene that was either congruent or incongruent in emotional valence with the prime. Participants responded whether the probe was emotionally positive or negative. Affective priming was demonstrated by shorter response latencies for congruent than for incongruent prime-probe pairs. This effect occurred when the prime was presented in the left visual field at a 300-msec prime-probe stimulus onset asynchrony, even when the prime and the probe were different in physical appearance and semantic category. This result reveals that the affective significance of emotional stimuli can be assessed early through covert attention mechanisms, in the absence of overt eye fixations on the stimuli, and suggests that right-hemisphere dominance is involved. PMID- 18405047 TI - Electrophysiological correlates of crossmodal visual distractor congruency effects: evidence for response conflict. AB - To investigate the basis of crossmodal visual distractor congruency effects, we recorded event-related brain potentials (ERP) while participants performed a tactile location-discrimination task. Participants made speeded tactile location discrimination responses to tactile targets presented to the index fingers or thumbs while ignoring simultaneously presented task-irrelevant visual distractor stimuli at either the same (congruent) or a different (incongruent) location. Behavioural results were in line with previous studies, showing slowed response times and increased error rates on incongruent compared with congruent visual distractor trials. To clarify the effect of visual distractors on tactile processing, concurrently recorded ERPs were analyzed for poststimulus, preresponse, and postresponse modulations. An enhanced negativity was found in the time range of the N2 component on incongruent compared with congruent visual distractor trials prior to correct responses. In addition, postresponse ERPs showed the presence of error-related negativity components on incorrect-response trials and enhanced negativity for congruent-incorrect compared with incongruent incorrect trials. This pattern of ERP results has previously been related to response conflict (Yeung, Botvinick, & Cohen, 2004). Importantly, no modulation of early somatosensory ERPs was present prior to the N2 time range, which may have suggested the contribution of other perceptual or postperceptual processes to crossmodal congruency effects. Taken together, our results suggest that crossmodal visual distractor effects are largely due to response conflict. PMID- 18405048 TI - Is task switching nothing but cue priming? Evidence from ERPs. AB - Recent findings suggesting that switch costs in the task-cuing paradigm are largely attributable to a change in the task-indicating cue have been interpreted in terms of a priming model of task-switch costs (Logan & Bundesen, 2003). According to this explanation, participants do not actually switch task sets, but merely use a cue-stimulus compound to disambiguate competing response tendencies associated with bivalent stimuli. Here, we report an event-related potential (ERP) experiment that provides evidence against this notion. In a paradigm with a 2:1 mapping between cues and tasks, we show that cue-switch and task-switch effects are dissociable on a neurophysiological level, indicating that task switching is more than a switch in the task-indicating cue. Moreover, a systematic analysis of the ERPs during the cue-stimulus interval suggests that updating processes can run in advance, before the stimulus is presented. PMID- 18405049 TI - Neurophysiology of successful encoding and retrieval of source memory. AB - Event-related potentials were recorded during encoding, to identify whether brain activity predicts subsequent retrieval of spatial source information, and during retrieval, to investigate the neural correlates of successful and unsuccessful spatial context recollection. The amplitude registered during encoding for study items that were later recognized and assigned a correct source judgment was more positive than the amplitude for recognized items given incorrect source judgments; this difference started 480 msec poststimulus, predominantly at central and anterior sites. During retrieval, the waveform was more positive from 250 to 1,600 msec poststimulus when the brain had retrieved episodic information successfully than when it had failed. These findings indicate that brain electrical activity recorded during the first presentation of an item within its context predicts the subsequent retrieval of the specific spatial context. During retrieval, brain activity differed quantitatively at anterior sites and qualitatively at posterior sites according to the accuracy of source memory. PMID- 18405050 TI - Motivational influences on cognitive control: behavior, brain activation, and individual differences. AB - What changes in brain activity are associated with changes in motivational state? The present study addressed this question by having participants perform a cognitive task (AX variant of the Continuous Performance Test; AX-CPT) under three different blocked motivational conditions (reward-incentive, penalty incentive, and baseline). Behavioral data showed that the incentive conditions modulated task performance, potentially by altering participants' cognitive control strategy. Neuroimaging data indicated that the reward condition was associated with a sustained increase in a primarily right-lateralized network that included parietal and prefrontal cortex. Additionally, individual differences were observed, such that activation in both reward-related brain regions and frontopolar cortex was linked to the degree of motivation-induced performance enhancement and to motivation-related personality variables. These results suggest that changes in motivational state may modulate performance through sustained activity in cognitive control regions and that the effect of incentives may be affected by the personalities of the participants. PMID- 18405051 TI - Subplate zone of the human brain: historical perspective and new concepts. AB - Subplate zone (SP) is prominent, transient laminar compartment of the human fetal cerebral wall. The SP develops around 13 and gradually disappears after 32-34 postovulatory weeks. The SP neurons can be found as late as nine postnatal months, while remnants of the SP neurons can be traced until adult age in the form of interstitial neurons of the gyral white matter. SP is composed of postmigratory and migratory neurons, growth cones, loosely arranged axons, dendrites, glial cell and synapses. The remarkable feature of the SP is the presence of large amount of extracellular matrix. This feature can be used for delineation of SP in magnetic resonance images (MRI) of both, in vivo and post mortem brains. The importance of SP as the main synaptic zone of the human fetal cortex is based on the rich input of ,waiting,< afferents from thalamus and cortex, during the crucial phase of cortical target area selection. SP increases during mammalian evolution and culminates in human brain concomitantly with increase in number and diversity of cortico-cortical fibers. The recent neurobiological evidence shows that SP is important site of spontaneous endogeneous activity, building a framework for development of cortical columnar organization. The SP which can be readily visualized on conventional and DTI (diffusion-tensor-imaging) MRI in vivo, today is in the focus of interest of pediatric neurology due to the following facts: (1) SP is the site of early neural activity, (2) SP is the major substrate for functional plasticity, and (3) selective vulnerability of SP may lead to cognitive impairment. PMID- 18405052 TI - Distinct origin of GABA-ergic neurons in forebrain of man, nonhuman primates and lower mammals. AB - In this mini-review we present recent data about origin of GABA-ergic (gama aminobutyric acid) neurons in the mammalian forebrain, including the diencephalon and telencephalon. The interest in GABA-ergic neurons, which in cerebral cortex mostly correspond to local circuit neurons (interneurons), has increased in the past decade. Many studies have shown that in lower mammals all hippocampal and almost all neo-cortical GABA-ergic neurons are born in the specific region named ganglionic eminence, and not locally in proliferative layers all around telencephalic vesicle. The ganglionic eminence, that represents a region with thick proliferative-subventricular layer in the ventral (basal) part of telencephalon, was classically thought to give neurons to basal ganglia and septal nuclei, whereas proliferative layers of dorsal telencephalon give neurons to cerebral cortex including hippocampus. It was thought that neurons migrate from proliferative layer to their target region following a radial orientation. However, data in lower mammals showed that this is the case only for glutamatergic principal cells, i.e. projection neurons. GABA-ergic neurons use long distance tangentional migration, parallel to pial surface to reach, from ganglionic eminence, their targeting layer in the cerebral cortex. Especially intriguing, but frequently neglecting, several studies suggest that mammalian evolution might use different developmental rules to provide GABA-ergic neurons to an expending brain. In this review we focus on specific events underlying GABA ergic neuron development in human and non-human primates. Disturbances of the GABAergic network are found in many neurological and psychiatric disorders, some of them might result from altered production or migration of these neurons during development. Therefore, it is crucial to understand human-specific mechanisms that regulate the development of GABA-ergic neurons. PMID- 18405053 TI - Music as an auditory stimulus in stroke patients. AB - Auditory stimulation increases mean blood flow velocity (MBFV) in the middle cerebral artery (MCA) in healthy individuals. Our aim was to monitor such changes in the affected MCA of patients with acute ischemic stroke (AIS). The study included 66 non-thrombolysed patients with AIS who were divided into groups according to National Institutes of Health Stroke Scale (NIHSS) score. Group I consisted of patients with NIHSS score 10 and group II with NIHSS score > or =11. Affected MCA was insonated with transcranial Doppler (TCD). MCA MBFVs were monitored during listening to music for 30 minutes. The first response of MBFV increase was measured as time (Tmax) and percentage of amplitude change (Amax). Pearson Chi-Square test was used. In 78.85% of patients there was a significant increase in MBFV compared to baseline values as a reaction to the music. There was no significant difference in Tmax or Amax between the two groups. However, a trend of longer Tmax was observed with every 2 NIHSS score increase. Music is an auditory stimulus in stroke patients and can be measured with TCD as MCA MBFV increase. Although our study showed no significant change of reaction time with the severity of stroke, a trend of prolonged Tmax was observed with NIHSS score increase. PMID- 18405054 TI - Changes of the corpus callosum in children who suffered perinatal injury of the periventricular crossroads of pathways. AB - There is a high incidence of periventricular leukomalacia, caused by hypoxia ischemia, in preterm infants. These lesions damage the periventricular crossroads of commissural, projection and associative pathways, which are in a close topographical relationship with the lateral ventricles. We explored to what extent abnormalities of echogenicity of the periventricular crossroads correlate with changes in size of the corpus callosum. Our study included nine infants (gestation from 26-41 weeks; birth weight between 938-4450 grams) with perinatal brain injury. Periventricular areas, which topographically correspond to the frontal, main and occipital crossroad, were readily visualized by cranial ultrasound scans, performed during the first two weeks after birth. Corpus callosum mediosagittal area measurements were performed using magnetic resonance images, acquired between the first and sixth postnatal month (postmenstrual age 40-49 weeks). We found a statistically significant correlation between the increased echogenicity in the crossroad areas and the decrease of the corpus callosum midsagittal area (p < 0.05). This supports the hypothesis that callosal fibers can be damaged, during growth through the periventricular crossroads of pathways. PMID- 18405055 TI - Cerebrospinal fluid markers in differential diagnosis of Alzheimer's disease and vascular dementia. AB - Alzheimer's disease (AD) and vascular dementia (VaD) are the two most common causes of dementia in old people. They remain difficult to differentiate in practice because of lack of sensitivity and specificity of current clinical diagnostic criteria. Recent molecular and cellular advancements indicate that the use of cerebrospinal fluid markers may improve early detection and differential diagnosis of AD. Our objective in this study was to determine diagnostic accuracy of three cerebrospinal (CSF) markers: total tau protein (t-tau), tau protein phosphorylated on threonine 181 (p-tau181) and tau protein phosphorylated on serine 199 (p-tau199). Using commercially available ELISA kits concentrations of t-tau, p-tau181 and p-tau199 were analyzed in 12 patients with probable AD, 9 patients with VaD and 12 NC subjects. The median levels of all three markers were significantly higher in AD group versus VaD and NC groups. However, when the sensitivity levels were set to 85% or higher, only t-tau and p-tau199 satisfied consensus recommendations (specificity more than 75%) when differentiating AD from VaD. In conclusion, our preliminary data on a small group of selected subjects suggest that the CSF t-tau and p-tau199 levels are useful markers for differentiating AD from VaD. PMID- 18405056 TI - Expression of 5HT-1A and 5HT-1B receptor genes in brains of Wistar-Zagreb 5HT rats. AB - By selective breeding, two sublines of rats with high or low activity of platelet serotonin (5HT) transporter (5HTt) have been developed (Wistar-Zagreb 5HT rats). Previous studies demonstrated significant differences between the sublines in the expression of platelet 5HTt at the level of both, mRNA and protein. Pharmacological studies showed marked alterations in brain 5HTt function, indicating differences in central serotonin homeostasis, although analysis of regional brain 5HTt gene expression did not show analogous differences. In this study, we searched for possible changes in the expression of the two central 5HT receptor subtypes: 5HT-1A and 5HT-1B, both participating in the regulation of brain 5HT transmission. Semi-quantitative RT-PCR, with three different housekeeping genes as internal standards, showed no differences in the levels of 5HT-receptor expression between the sublines. Results suggest that constitutional alteration of 5HT homeostasis, induced by selective breeding for the extremes of platelet 5HTt activity, did not cause measurable changes in the expression of central 5HT-1A (hippocampus) and 5HT-1B (striatum) receptors in the mentioned rat sublines under physiological conditions. PMID- 18405057 TI - Transventricular and transpial absorption of cerebrospinal fluid into cerebral microvessels. AB - It is generally accepted that volume of cerebrospinal fluid (CSF) is secreted in brain ventricles and flows to subarachnoid space to be absorbed into dural venous sinuses or/and into lymphatics via perineural sheats of cranial nerves. Since 99% of CSF volume is water, in experiments on cats 3H-water was slowly infused into lateral ventricle and found that it does not flow to subarachnoid space but that it is rapidly absorbed transventricularly into periventricular capillaries. When 3H-water was infused in cortical subarachnoid space, it was absorbed locally into cerebral capillaries via pia mater. On the contrary, when macromolecule 3H-inulin is applied in CSF it is very slowly eliminated in bloodstream, and, with time, is carried by systolic-diastolic pulsations and mixing of CSF bidirectionally along CSF system. Thus, CSF volume (water) is absorbed rapidly into adjacent cerebral capillaries while inulin is distributed bidirectionally due to its long residence time in CSF Previously, the macromolecules have been used to study CSF volume hydrodynamics and with this misconception of CSF physiology arose. PMID- 18405058 TI - Mitochondria morphology and DNA content upon sublethal exposure to beta-amyloid(1 42) peptide. AB - Brains affected by Alzheimer's disease (AD) show a large spectrum of mitochondrial alterations at both morphological and genetic level. The causal link between amyloid beta peptides (AP) and mitochondrial dysfunction has been established in cellular models of AD using Abeta concentrations capable of triggering massive neuronal death. However, mitochondrial changes related to sublethal exposure to Abeta are less known. Here we show that subtoxic, 1 microM Abeta(1-42) exposure does not change the mitochondrial shape of living cells, as visualized upon the uptake of the non-potentiometric fluorescent probe Mitotracker Green and enhanced yellow fluorescent protein (EYFP)-tagged cytochrome c oxidase expression. Immunolocalization of oxidative adducts 8 hydroxy-2'-deoxyguanosine, 8-hydroxyguanine and 8-hydroxyguanosine demonstrates that one-micromolar concentration of Abeta(1-42) is also not sufficient to elicit dramatic qualitative changes in the RNA/DNA oxidative products. However, in comparison with controls, semi-quantitative analysis of the overall mitochondrial mass by integrated fluorescence intensity reveals an ongoing down-regulation in mitochondrial biosynthesis or, conversely, an enhanced autophagic demise of Abeta treated cells. Furthermore, a significant increase of the full-length mitochondrial DNA (mtDNA) from Abeta-treated versus control cells is found, as measured by long range polymerase chain reaction (PCR). Such up-regulation is accompanied by extensive fragmentation of the unamplified mtDNA, probably due to the detrimental effect of Abeta. We interpret these results as a sequence of compensatory responses induced by mtDNA damage, which are devoted to repression of oxidative burst. In conclusion, our findings suggest that early therapeutic interventions aimed at prevention of mitochondrial oxidative damage may delay AD progression and help in treating AD patients. PMID- 18405060 TI - Prevalence of mood dysfunction in epilepsy patients in Croatia. AB - Fifty consecutive and consenting epilepsy patients from the Zagreb Epilepsy Center were examined for the presence of depressive symptoms using the Beck Depression Inventory (BDI). This questionnaire has been previously validated for use in the Croatian population. Mean age of the patients was 30.8 +/- 13.5 years, 60.4% were females. Majority of them were employed (72.9%) and single (62.5%), and 35.4% had a university degree. Most of them had complex partial seizures (n=40, 80%), and 6 (12%) were diagnosed with idiopathic generalized epilepsy. Assessment with the BDI showed that 33.3% of patients had recent depressive symptoms: 6.3% had mild depressive symptoms, 8.4% moderate and 18.6% severe depressive symptoms. Three patients (6.4%) attempted suicide in the past, two of them had current suicidal ideation, and all of them were severely depressed. This is the first and preliminary study assessing mood dysfunction in epilepsy patients in Croatia. Increased prevalence of depression in epilepsy patients suggests specific approach and need for early treatment. PMID- 18405059 TI - Expression pattern and functional analysis of mouse Stam2 in the olfactory system. AB - Gene trap mutant mice Stam(gt1Gaj) were investigated in order to elucidate in vivo function of Stam2 (signal transducing adaptor molecule 2) gene, which was in vitro implicated in sorting cargo marked by monoubiquitination toward degradation in the lysosomes. The expression analysis showed high Stam2 expression in the brain including the regions related to olfaction, and in the olfactory epithelium, but not in the respiratory part of nasal mucosa. To test mouse olfaction, ability to find chocolate hidden under the sawdust in the cage was examined. When food was given ad libitum before trials, mutants needed more time and failed more frequently to find the chocolate. In contrast, when the mice were fasted overnight before trial, there were no differences between mutants and wild type mice. No changes in morphology of olfactory mucosa were observed. The obtained results showed the existence of phenotype differences between mutants and wild type mice. However, different results of two approaches aimed to test olfaction, with and without food deprivation, currently do not enable to assign the particular function of Stam2 to olfaction. This emphasizes how slight modification of experimental setup in behavioural testing can cause important differences on the obtained results. PMID- 18405061 TI - Dream recall after night awakenings from tonic/phasic REM sleep. AB - Eleven healthy subjects, 9 females and 2 males aged 21-23, were submitted to all night polygraphic recording and awaken in REM (Rapid Eye Movements) sleep, randomly upon tonic or phasic REM. Immediately upon awakening subjects were asked about possible dreaming according to the standardized questionnaire. Seventy seven dreams, i.e. 79% of all 97 REM awakenings, were reported and analyzed. There were no significant differences in reported frequency of dreamings after awakening, mood and dream content due to phasic/tonic REM sleep. Dreams from phasic REM were a bit more colorful. Predictor of morning remembering of dreams was meaninglessness, not meaningfulness of dreams, and, in lesser extent, good mood, colorfulness, dreams with words and phasic REM sleep. PMID- 18405062 TI - Hyperserotonemia in autism: the potential role of 5HT-related gene variants. AB - Increased platelet serotonin level (PSL) has been consistently found in a portion of autistic patients. Suggested mechanisms for hyperserotonemia in autism have been increased synthesis of serotonin (5HT) by tryptophan hydroxylase (TPH), increased uptake into platelets through 5HT transporter (5HTt), diminished release from platelets through 5HT2A receptor (5HT2Ar) and decreased metabolism by monoamine oxydase (MAOA). The allelic influence of genes, encoding the mentioned 5HT elements, on PSL was investigated in 63 autistic subjects. Our study shows that 5HTt-LPR and -1438AG 5HT(2Ar) genotypes did not significantly affect PSL. However, significantly higher PSLs were observed in subjects with "cc" genotype of a218c TPH and subjects with "4" genotype of uVNTR MAOA. In addition, when TPH-cc and MAOA-4 were combined as "high 5HT" genotypes, a correlative increase in PSL was observed with the increase in the number of "high 5HT" genotypes. These results suggest a possible synergistic effect of genes regulating 5HT synthesis/degradation in dysregulation of the peripheral 5HT homeostasis of autistic patients. PMID- 18405063 TI - Femininity and masculinity across the menstrual cycle: a relation to mate value. AB - Numerous studies have shown that menstrual cycle related variations in sex hormones influence various cognitive processes. These shifts are considered as the evidence for a hormone-mediated adaptive design underlying human mating motivation. In a series of related studies we have shown that (i) femininity does not vary across the menstrual cycle, whereas masculinity is the most pronounced during the fertile period, (ii) masculinity, but not femininity, predicts shifts in spatial cognition across the menstrual cycle, and (iii) women with different positions on masculinity and femininity dimensions differ in their self-perceived mate value. These results suggest that (i) there might be a hormone mediated psychological mechanism making a woman more assertive and dominant during a short time-window when the conception is likely, (ii) menstrual cycle related shifts in cognitive abilities and mating motivation might have a common hormonal mechanism, and (iii) women's mate value (and indirectly her reproductive success) depends upon both feminine and masculine traits. PMID- 18405064 TI - Lithium effect on glutamate induced damage in glioblastoma cells. AB - Lithium, besides mood stabilization, might be involved in neuroprotection. Previously we have found that the treatment with lithium increased the levels of p21(WAF/Cip1 and survivin in human glioblastoma A1235 cells. The aim of the present study was to examine the cytotoxic effect of glutamate on these cells, and to determine whether lithium can protect A1235 cells against toxic effects of glutamate. Cytotoxicity of glutamate was examined by spectrophotometric MTT assay, while the expression of apoptosis related genes was examined by Western blot method. Glutamate was excessively cytotoxic for A1235 cells only in concentrations higher than 100 mM. It did not induce apoptosis, but rather suppressed survivin expression and increased the level of p21(WAf/Cip1). Pretreatment with lithium (2 mM) partially reverted change in survivin expression induced by glutamate, suggesting that lithium may have beneficial effect on glutamate induced cell damage in glioblastoma cells. PMID- 18405065 TI - The effect of repeated daily measurements on paw withdrawal latencies in Hargreaves test. AB - The hypothesis that repeated measurements during 4 subsequent days affect withdrawal latencies in Hargreaves test was investigated. Paw withdrawal latencies to radiant heat were determined in the control, tramadol or saline group of male Wistar rats. The control group (N=10) had no treatment. Tramadol group (N=7) and saline group (N=7) received one daily intraperitoneal injection of tramadol (15 mg/kg) or saline (0.9% NaCl), respectively. A significant decline in withdrawal latencies was observed in the control group on the day 2 to day 4, when compared to day 1 (p < 0.05 Bonferroni test). In the saline and tramadol groups, latencies remained stable from day 1 to day 4. During the entire testing period withdrawal latencies were 27-50% longer in tramadol group (p < 0.05 ANOVA) compared with the saline group. When compared to the control group, the effect of tramadol, was noted from the second to forth day (p < 0.01 Bonferroni test), but not on the first day. Finally, a tendency to decrement in withdrawal latencies existed on day 1 in the saline group compared with control group, but this difference does not reach significance. We conclude that one day of training affect withdrawal latencies in the Hargreaves test. PMID- 18405066 TI - Increased permeability of the blood-brain barrier following administration of glyceryl trinitrate in common carp (Cyprinus carpio L.). AB - Nitric oxide (NO) has been implicated in the pathogenesis of migraine and treatment with its exogenous donor glyceryl trinitrate (GTN) represents widely accepted experimental "migraine model". In this study, glyceryl trinitrate was administered intraperitoneally to carps, serum nitrite and nitrate levels were determined, permeability of blood-brain barrier was investigated, and histological changes of brain tissue were analyzed. Serum nitrite and nitrate levels displayed characteristic biphasic pattern with moderate initial increase and maximal terminal increase, suggesting the GTN-induced endogenous NO synthesis. Increased permeability of the blood-brain barrier in GTN-treated animals was determined based on Evans blue capillary leakage into the brain tissue. Histological analysis revealed changes consistent with vasodilatation and oedema. Our study strongly supports the importance of the NO role in the pathogenesis of migraine attacks and increase in blood-brain barrier permeability during the attack. The study has also provided evidence that this mechanism of action is conserved to the lower vertebrate. PMID- 18405067 TI - West syndrome with periventricular leukomalacia: ten-year clinical study. AB - The aim of the study was to evaluate magnetic resonance imaging (MRI) findings in infants with periventricular leukomalacia (PVL) and West syndrome (WS) and determine the neurodevelopmental outcome in children with West syndrome and PVL. Ultrasound and brain MRI were performed in 37 infants with recognized PVL. PVL was categorized according to De Vries, whereas West syndrome was categorized according to International League Against Epilepsy 1989. West syndrome in our patients developed during the first 2 years of life. The most common interictal abnormality was hypsarrhythmia. All, except two patients had delayed development and various degrees of mental retardation. The most characteristic neuroimaging findings were major reduction in cerebral cortical gray matter volume, reduction in the volume of brain myelin, and delayed myelination. These findings may explain the anatomical association between the West syndrome onset and PVL and intellectual and cognitive deficit in premature infants with PVL. PMID- 18405068 TI - Serum cholesterol concentrations in suicidal and non-suicidal male patients suffering from persistent delusional disorder. AB - Suicidal behavior is a major health risk in psychiatric disorders, especially in affective and psychotic disorders. The neurobiology of suicidal behavior is still unclear. Suicidality has been related to a reduced cholesterol levels. The aim of this study was to evaluate serum cholesterol concentrations in suicidal and non suicidal men suffering from persistent delusional disorder and in healthy volunteers. Results showed that serum cholesterol concentrations were significantly lower in suicidal than in non-suicidal patients and healthy controls. Also, level of psychopathology (measured by Brief Psychiatric Rating Scale) is significantly marked in the group of suicidal patients, which indicates the importance of detecting some clinical symptoms in patients with persistent delusional disorder in order to prevent suicidal behavior. PMID- 18405069 TI - Platelet serotonin and monoamine oxidase in Alzheimer's disease with psychotic features. AB - Post mortem brain studies indicate that alterations in serotonergic and catecholaminergic systems might be associated with Alzheimer's disease (AD). The aim of the study was to determine serotonin (5-HT) levels and monoamine oxidase type B (MAO-B) activity in platelets of psychotic and non-psychotic patients with AD, established according to the NINCDS-ADRDA and DSM-IV-TR criteria. Cognitive impairment and psychotic features were evaluated using Mini Mental Status Examination and Neuropsychiatric Inventory. Platelet 5-HT concentration and MAO-B activity were determined spectrofluorimetrically in 116 (51 male, 65 female) healthy subjects and 70 psychotic (10 male, 60 female) and 151 non-psychotic (32 male, 119 female) patients. Psychotic and non-psychotic female and psychotic male patients had significantly lower platelet 5-HT concentration than corresponding sex matched control subjects. Platelet MAO-B activity was significantly increased in both male and female non-psychotic patients compared to the sex matched controls. Non-psychotic female patients had significantly higher platelet MAO-B activity than psychotic female patients. Our data suggest that platelet MAO-B activity, but not platelet 5-HT concentration, could differentiate between psychotic and non-psychotic subtypes of AD. PMID- 18405070 TI - Nucleolar protein 1 (Nol1) expression in the mouse brain. AB - Nucleolar protein 1 (Nol1) is a cell cycle dependent gene highly expressed in proliferating tissues. In order to test whether Nol1 could be used as a marker of dividing neural stem cells within mouse brain, Nol1 expression was analyzed using mouse carrying a gene trap modification of Nol1 gene. High Nol1 expression was found within the hippocampus, olfactory bulb, cerebral and cerebellar cortex. Nol1 was expressed not only in the dividing cells within the brain, but as well in the postmitotic neurons. This suggested a general role of Nol1 in assembling of ribosomes in cells with high protein production. PMID- 18405071 TI - Alcohol dependence and polymorphisms of serotonin-related genes: association studies. AB - Variations in 5HT-related genes contribute to the alterations of serotonergic neurotransmission, which is implicated in the etiopathology of alcoholism. In this preliminary study we have tested polymorphisms of genes involved in 5HT transport and turnover for their association with alcohol dependence. A case group of males with type 2 alcoholism (N=59) and a control group of healthy males (N=282), both of Croatian origin, were analyzed for the frequency distribution of polymorphisms in 5HT transporter (5HTT-VNTR2, 5HTT-LPR), monoamine oxidase A (MAOA-uVNTR) and B (MAOB-A/G) and tryptophan hydroxylase 1 (TPH1 A218C) and 2 (TPH2 G-703T) genes. An increase in the frequencies of 10-repeat allele (p = 0.010; OR = 1.73; 95% CI = 1.14-2.60) and 10/10 genotype (p = 0.006; OR = 2.57; 95% CI = 1.32-5.00) of the 5HTT-VNTR2 polymorphism was found in alcoholic patients. No differences between case and control groups were observed for the other tested polymorphisms. Present results support earlier studies implicating the role of 5HTT gene in alcoholism. The increase of sample size (in progress) is expected to enable search of more subtle differences, as well as re-evaluation of these preliminary findings. PMID- 18405072 TI - Fluid perfusion as a method of cerebrospinal fluid formation rate--critical appraisal. AB - The aim of the study was to evaluate whether or not cerebrospinal fluid formation rate (Vf) calculated according to the equation of Heisey et al., truly show the produced cerebrospinal fluid. For this reason Vf was simulated (40.6 microL/min) by an infusion pump in a plastic cylinder and the evaluation was done by comparing the results obtained between the calculated Vf and the simulated one. In both cases the result should be the same (40.6 micro/min). Other types of experiments were carried out by ventriculocisternal perfusion (92.4 microL/min) on anaesthetized and sacrificed cats. If the equation is correct, the calculated Vf for sacrificed animals should be zero, because there is no Vf in dead animals. The fact that the calculated Vf (46.5 microL/min) in the plastic cylinder was different (p < 0.0001) from the simulated one (40.6 microL/min) and that Vf was calculated even for dead animals (3-5 microL/min) clearly shows the that perfusion method may not be an accurate method for determination of Vf. PMID- 18405073 TI - Temporal lobe volume in disorders with psychotic features. AB - Since early neuropathological findings, temporal lobe has been related to the pathophysiology of some disorders with psychotic features. The aim of this study was to compare temporal lobe volumes and asymmetry differences in patients with schizophrenia, schizoaffective and bipolar disorders, disorders that cover the whole psychotic spectrum. Temporal lobe volumes were estimated using high resolution magnetic resonance imaging in 60 subjects, 15 subjects in each patient and one healthy volunteer (control) group. There are no statistically significant differences in temporal lobe volumes among patient and control groups. Comparison of left and right temporal lobes shows that left temporal lobes are smaller than right temporal lobes, however this difference reaches statistical significance only in groups of patients with schizoaffective and bipolar disorders. Overall temporal lobe volume may be less informative in respect to neuropathology of disorders with psychotic features than volumes of specific temporal lobe structures, in particular medial temporal lobe structures. PMID- 18405074 TI - Neurodevelopmental outcome in children with periventricular leukomalacia. AB - The purpose of this study was to question the correlation of different grades of periventricular leukomalacia (PVL) and subsequent neurodevelopmental outcome. In a prospective study we followed 52 preterm infants. Infants were divided into three groups according to their cranial ultrasound findings of PVL (De Vries classification). Seventeen children had PVL 1, 20 children had PVL 2, and 15 children had PVL 3. All 15 (100%) children with PVL 3 developed cerebral palsy with additional visual perceptual dysfunctions and epilepsy. Children with PVL 1 had high frequency of mild neuromotoric delay and visual impairment. PVL 2 and 3 have great predictive value for subsequent severe neurodevelopmental disorder which refers to cerebral palsy, different cognitive deficits, vision impairment and epilepsy. We have determined that due to high frequency of visual impairment and epilepsy we need to include neurophysiologic examinations very early in children with PVL lesions. PMID- 18405075 TI - Foetal Leydig cells and the neuroendocrine system. AB - It has been shown that adult human Leydig cells express a number of neuroendocrine markers, and, therefore, could be considered as a part of the neuroendocrine system in the adult. A limited number of studies have dealt with the dynamics of development of human foetal Leydig cells, whereas studies on their neuroendocrine nature are still extremely rare. Therefore, the aim of our study was to investigate the development of human foetal Leydig cells in different weeks of gestation (wg) and to check if these cells express certain markers characteristic of the diffuse neuroendocrine system (DNS). Qualitative, quantitative histological studies and immunohistochemical analyses of human foetal testicular tissue have been performed. According to our data, Leydig cells formed a dynamic population of cells within the interstitum of testes in the period between 15 and 36 wg. The total number of Leydig cells of human foetal testes changed through different stages of gestation by means of 'pulsatile' dynamics (most likely, by following the variable level of gonadotropins). At early stages of development (15-17 wg) immunohistochemical reactions for the expression of neuron specific enolase (NSE) were positive within sex cords and between them, in the interstitum. Pro-spermatogonia in the sex cords were positive, as well as elongated spindle-shaped cells of the interstitum (very likely, precursors of Leydig cells). During the later stages of development (28 36 wg), excluding the pro-spermatogonia, the interstitial Leydig cells were also positive. The results of the immunohistochemical analyses (the expression of NSE) confirmed the hypothesis that human foetal Leydig cells were of neuroendocrine nature. PMID- 18405076 TI - Structural basis of developmental plasticity in the corticostriatal system. AB - Previous studies have shown that in developing monkey corticostriatal fibres terminate around striatal cytoarchitectonic compartments--cell islands, showing transfiguration around 105th embryonic day (E105) of gestation. In the present study we have analyzed these striatal cytoarchitectonic islands and acetylcholinesterase (AChE) rich patches in the developing human brain considering them as structural indicators of the development of the corticostriatal pathways. Postmortal brain tissue of 27 fetuses and prematurely born infants, ranging from 11-34 postovulatory weeks (POW) whose deaths were attributed to non neurological causes, were processed by Nissl method, AChE histochemistry and imunocytochemical technique (synaptophysin). All specimens are part of the Zagreb Neuroembryological Collection. Initial AChE patches, presumably corresponding to the dopaminergic islands, were seen as early as 10 POW whereas cytoarchitectonical cell islands were not observed until 14 POW The main developmental change occurs between 20-24 POW when AChE negative cell poor zones develop around cell islands. This transient AChE pattern of striatal organization reaches its peak around 28 POW being most prominent along lateral border of putamen. In one case of periventricular hemorrhagic lesion with premortem survival period we have found reorganization of AChE patches in the putamen which indicates structural plasticity of corticostriatal pathways. In conclusion we propose that cell poor zones serve as waiting compartments for growing corticostriatal fibers which approach striatum through subcallosal bundle and external capsule. The period of the existence of striatal compartments (14-30 POW) is a sensitive period for structural plasticity and vulnerability after periventricular lesions. PMID- 18405077 TI - Quantitative analysis of basal dendritic tree of layer III pyramidal neurons in different areas of adult human frontal cortex. AB - Large long projecting (cortico-cortical) layer IIIc pyramidal neurons were recently disclosed to be in the basis of cognitive processing in primates. Therefore, we quantitatively examined the basal dendritic morphology of these neurons by using rapid Golgi and Golgi Cox impregnation methods among three distinct Brodmann areas (BA) of an adult human frontal cortex: the primary motor BA4 and the associative magnopyramidal BA9 from left hemisphere and the Broca's speech BA45 from both hemispheres. There was no statistically significant difference in basal dendritic length or complexity, as dendritic spine number or their density between analyzed BA's. In addition, we analyzed each of these BA's immunocytochemically for distribution of SMI-32, a marker of largest long distance projecting neurons. Within layer IIIc, the highest density of SMI-32 immunopositive pyramidal neurons was observed in associative BA9, while in primary BA4 they were sparse. Taken together, these data suggest that an increase in the complexity of cortico-cortical network within human frontal areas of different functional order may be principally based on the increase in density of large, SMI-32 immunopositive layer IIIc neurons, rather than by further increase in complexity of their dendritic tree and synaptic network. PMID- 18405079 TI - Mental lexicon and derivational rules. AB - Lexical decision task in an event-related potential experiment was used in order to determine the organization of mental lexicon regarding the polimorphemic words: are they stored as unanalyzable items or as separate morphemes? The results indicate the later: while monomorphemic words elicit N400 component, usually related to lexical-semantic processing, prefixed words and prefixed pseudo-words elicit left anterior negativity (LAN), usually related to grammatical (morphosyntactic) processes. These components indicate that the speakers apply grammatical (i.e., word-formation) rules and combine morphemes in order to obtain lexical meaning of the prefixed word. PMID- 18405078 TI - Serum gangliosides in patients with brain tumors. AB - In order to determine possible differences in serum gangliosides content and composition before and after surgical removal of tumor, gangliosides isolated from preoperative and postoperative sera of patients with brain tumors were analyzed. Serum samples were collected from patients with glioblastoma, meningioma, acoustic neurinoma, haemangioma, oligodendroglioma and astrocytoma, one week before and one week after surgical removal of the tumor. Serum gangliosides were qualitatively and quantitatively analyzed by high performance thin layer chromatography and laser densitometry. Results showed changes of total gangliosides concentrations in analyzed postoperative sera comparing to preoperative sera. There was not a significant difference in ganglioside pattern of preoperative vs. preoperative sera. However, a postoperative decreased proportion of ganglioside GD3 was observed in sera derived from patients with complete tumor removal. The results of this study indicate that comparative quantitative and compositional analysis of both preoperative and postoperative serum gangliosides may provide useful information concerning tumor progression, surgical success and prognosis. PMID- 18405080 TI - Vasodilatory prostaglandins in perinatal hypoxic brain damage. AB - Prostaglandin (PGE2 and PGI2) synthesis was determined in the cerebrospinal fluid (CSF) and serum of 19 hypoxic neonates at the age of 5-96 hours by using Enzyme Linked Immunosorbent Assay (ELISA) method. Control group consisted of 8 children of the same age whose samples were taken due to initial suspicion of neonatal meningitis. The prostaglandin concentrations in CSF were correlated with initial hypoxic-ischemic encephalopathy (HIE) stage and neurological findings of patients at the age of 12 months. The values of PGE2 and PGI2 in the CSF of children with perinatal hypoxia (PNH) were significantly higher than in the children from the control group. The values of PGI2 in serum were significantly higher than in "CSF" of patients with PNH. Although average values of PGE2 and PGI2 in the liquor were higher in children with advanced stage of HIE, the differences between different stages were not statistically significant. We did not find any significant correlation between average concentrations ofprostaglandins and neurological findings of the 12-month-old children. PMID- 18405081 TI - Glycosphingolipid expression in cerebrospinal fluid of infants with neurological abnormalities: report of three cases. AB - The aim of this study was to analyse glycosphingolipid expression in cerebrospinal fluid (CSF) from one idiopathic West syndrome (IWS) infant, one with Reye like syndrome, and one with congenital hydrocephalus, in comparison to control group (n=7) using highly sensitive thin-layer chromatography immunostaining methods. Gangliotetraose-series gangliosides (acidic glycosphingolipids) were not detected in CSF of infant with idiopathic West syndrome and infant with congenital hydrocephalus. CSF of infant with IWS showed traces of neolacto-tetraose ganglioside fractions, which were absent in all other CSF examined. In addition, lactosylceramide fraction, and one ceramide fraction were highly expressed only in IWS CSF These results confirmed previously described lack of gangliotetraose-series gangliosides in IWS patient and for the first time is described increased expression of neolacto-series glycosphingolipids in IWS patient. Since follow up until the age of five years showed almost normal IWS patient psychomotor development, the discribed shift of glycosphingolipid expression may implicate on transient inhibition of specific glycosyl transferases in the age of seven months. PMID- 18405082 TI - Giant choroid plexus cyst as an accidental finding in an older man. AB - Choroid plexus cysts (CPC) are usually found at the end of the second trimester of pregnancy. Sometimes they can be accidentally and found on prenatal ultrasound examinations. Vast majority of CPC resolve spontaneously by 28th weeks gestation. In the older aged group the choroid plexus cysts are extremely rare pathomorphologic medical entity. Since they are almost always asymptomatic, they are therefore accidentally found on brain magnetic resonance (MR) or computed tomography (CT) scans. They are usually located in the lateral ventricles and measure around 2 cm in diameter. We present a case of a 75-year-old male with a giant choroid plexus cyst whose leading symptom was excruciating headache refractory to previous conservative therapy. He underwent surgery when osteoplastic craniotomy was performed with cyst fenestration and ablation. His recovery was uneventful with total regression of headaches. Reviewing the recent literature we did not find such a case considering the patients age and the size of the choroid plexus cyst. PMID- 18405083 TI - Magnetic resonance spectroscopy and measurement of tau epitopes of autopsy proven sporadic Creutzfeldt-Jakob disease in a patient with non-specific initial EEG, MRI and negative 14-3-3 immunoblot. AB - Limited potential of electroencephalogram (EEG), magnetic resonance images (MRI) and cerebrospinal fluid (CSF) test for 14-3-3 protein in the clinical diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) resulted in developments in diagnostic premortem tehniques. Recent studies provided evidence that magnetic resonance spectroscopy (MRS) and measurement of total-tau (T-tau) and phospho-tau (P-tau) may be useful to identify patients with CJD. We combined detected metabolic changes in the brain by MRS and measured T-tau and tau-pT181 by ELISA, and tau-pT231 by Westernblot in a patient with autopsy proven sCJD. Our results show that in contrast to negative CSF 14-3-3 protein, nonspecific EEG and MRI, MRS revealed metabolic alterations in regions of the brain that has appeared normal on MRI, and tau tests has shown measurable levels of phosphorylated and non-phosphorylated isoforms in CSF. We conclude that rapidly progressive dementia with negative 14-3-3 test and non-specific initial EEG and MRI must still be considered in the differential diagnosis of the sCJD. Combination of serial functional MRI along with MRS study and measurement of tau ratio could improve the early diagnosis of sCJD. The current case is the first attempt to study results of the use of MRS and tau tests in a case of sCJD with diagnostic dilemma. PMID- 18405084 TI - Magnetic resonance imaging and magnetic resonance spectroscopy in a patient with amyotrophic lateral sclerosis and frontotemporal dementia. AB - Magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) have been investigated in a single neurodegenerative disease manifesting as either amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD) alone, but have not been examined in combined disorders such as ALS with FTD (ALS-FTD). To our knowledge, this study is the first attempt to demonstrate relationship between MRI abnormalities and MR spectroscopic metabolite changes of the motor cortex, frontal white matter and corticospinal tract in a patient with the diagnosis of ALS with probable upper motor neuron signs (ALS-PUMNS) and FTD. Patient presented underwent MRI of the brain and MRS. The ratio of N acetylaspartate (NAA) to creatine (Cr), choline to Cr, myo-inositol (ml) to Cr and glutamate-glutamine (Glx) to Cr were derived from peak area measurement. Spectra from the right motor cortex, frontal white matter and corticospinal tract were obtained. MR images were evaluated for sulcus centralis enlargement, corticospinal tract hyperintensity and frontal lobes atrophy. Spectra showed reduced NAA/Cr and Glx/Cr ratio, yet the ratio of Cho/Cr exhibited significant elevation. MR images revealed sulcus centralis enlargement, high signal intensity of corticospinal tract and atrophy of both frontal lobes. Proton spectroscopic metabolic changes in a current patient fully correlate with previously reported MRS metabolic changes in ALS alone. Surprisingly, normal ml (glial marker) values have been found in almost all measured voxels of interest except in the frontal white matter. These findings differ from the previous findings in ALS or FTD alone. In conclusion, these findings support the concept that ALS, FTD and ALS FTD may represent different manifestations of a single pathological continuum. PMID- 18405085 TI - Dynamic in prescribing antipsychotic drugs during five year period (2001-2005) in the Psychiatric Hospital Vrapce, Zagreb, Croatia. AB - Aim of this study was to determine changes in prescribing antipsychotics during 5 year period in Psychiatric Hospital VrapEe. Data about type of antipsychotic medication, ward and gender were collected for all patients receiving antipsychotics on 1st of October. During 5 year period decrease in prescribing classical antipsychotics was observed while prescribing of atypical antipsychotics has shown increase. There was an increase in number of patients treated with combination of antipsychotics, while number of patients treated with clozapine remained the same. It was noticed that female patients were more often treated with atypical antipsychotics. Data for forensic and emergency ward was analyzed separately and trends similar to hospitals were noticed here. Rationalizing use of antipsychotics can decrease cost of treatment, decrease negative effects of antipsychotics and consequently improve the treatment. Through systematic studies of this type positive progress and changes in the prescribing of antipsychotics are possible. PMID- 18405086 TI - The art, science and commerce of medical tourism. PMID- 18405087 TI - A prospective study of medical students' perspective of teaching-learning media: reiterating the importance of feedback. AB - To enhance successful communication, medical teachers are increasingly using teaching-learning media. To determine medical students' perception of three such media (blackboard, overhead projector, and slides), and to generate recommendations for their optimal use, a prospective questionnaire-based study was carried out among 7th semester medical students of the University College of Medical Sciences and Guru Teg Bahadur Hospital, Delhi. Students made a forced choice between: (1) The three media on 8 questions regarding their advantages and disadvantages and (2) four aspects of a lecture (teaching-learning media, topic, teacher and time of day) regarding which made the lecture most engaging. Resulting data was analysed by Chi-square and Fisher's exact tests. Chalk and blackboard was rated as best in allowing interaction and helping recall (p<0.001 each). The overhead projector was best in providing information pointwise (p<0.001; 67 students, 89.3%, considered this an advantage). More subject matter could be covered per lecture (p=0.001; 58 students, 77.3%, considered this a disadvantage). Slides were best in imparting clinical details (p=0.004), but were sleep inducing (p<0.001). The teacher's style of instruction was most important in making the lecture engaging (p<0.001). The teacher's role in the learning process is important. Students enjoy the slow pace and interaction allowed by blackboard, pointwise information presented by the overhead projector, and the clinical details a slide can provide. The results suggest that the lecture could best be a combination of two or more teaching-learning media. Students' interaction should be encouraged whatever the media used. PMID- 18405088 TI - Comparative evaluation of perimenopausal abnormal uterine bleeding by transvaginal sonography, hysteroscopy and endometrial biopsy. AB - To evaluate the causes of abnormal uterine bleeding in perimenopausal women and to achieve the greatest diagnostic accuracy with the least risk, a cross sectional study was done among 85 patients between the ages 40 and 55 years. Transvaginal sonography, hysteroscopy and histopathological examination of endometrium were done in all the cases. Kappa statistics was calculated to show the agreement between the different investigations. Considering histopathological report to be the gold standard, for diagnosis of hyperplastic endometrium, transvaginal sonography and hysteroscopy showed fair agreement (k=0.34) and good agreement (k=0.51) and hysteroscopic diagnosis of polyp showed strong agreement (k=0.81). Sensitivity, specificity, positive predictive value, negative predictive value for diagnosis of hyperplastic endometrium were 43.75%, 95.65%, 70% and 88% respectively whereas in polyp by transvaginal sonography they were 50%, 89.16%, 100%, 98.67% respectively; by hysteroscopy these were 50%, 95.78%, 70%, 90.36% respectively in hyperplasia and 71.43%, 100%, 100%, 94.67% respectively in polyp. Transvaginal sonography is most important for diagnosis of anatomical lesion. Hysteroscopy is most specific and sensitive for diagnosis of polyp but less specific for endometrial hyperplasia. PMID- 18405089 TI - Control and prevention of rat fever (Leptospirosis) outbreak in six villages of Raichur district, Karnataka. AB - The study describes an attempt to record leptospirosis in Raichur taluk/district, in Karnataka, by serological test such as microscopic test and IgM by ELISA. A house to house survey was done in six villages of Krishna river basin. Cases were interviewed regarding chief complaints like, fever, headache and oliguria, age, sex and treatment given with diclofenac, doxycycline, tetracycline and vitamin B complex. A total of 1516 cases were treated. The age of the patients ranged between 10 and 71 years. Most of them were agricultural workers. Out of 15 blood samples 12 samples sent to DHO Office, 3 were negative for leptospirosis, the density of rats were increased in all villages, the mode of transmission was food and water contamination, duration of incubation period was 5 to 10 days. Health education, for the community was undertaken to take measures against rats. Causative agent may be Leptospirosis interrogans, 3 cases were referred to Civil Hospital, Raichur and Navodaya Medical College, Raichur. Community participation was excellent. This study showed the presence of leptospirosis among the community of 6 villages, the disease is posing health hazards for agricultural workers in Raichur taluka villages. PMID- 18405090 TI - Community interventions against depression. AB - Depression appears to be the common psychiatric dosorder in any given community. Depression in different forms may affect 10% of the population at any given time. The paucity of mental health power has made people to ignore the presence of depression and its impact on individual's capacity of functioning. If we have to plan community based interventions some strategies are to be adopted. In primary healthcare systems short training of all categories of personnel in PHC and regular supply of free medicines are essential. With the experiences of general practitioners and their involvement, patients with depression can be approached for help. So also school and college teachers, trained counselors, religious and spiritual leaders can be involved in the processes. Family members can take care of patients. Stress management techniques, helpline, crisis intervention can be other methods to help the patients suffering from depression. PMID- 18405091 TI - Care of nasal airway to prevent orthodontic problems in children. AB - Obstruction in the upper respiratory passage is a cause of mouth breathing. A mouth breather lowers the tongue position to facilitate the flow of air in to the expanding lungs. The state of equilibrium of forces inside themouth is disturbed, thereby resulting altered soft tissue force acting on the bones in between. The resultant effect is maldevelopment of the jaw in particular and deformity of the face in general. Setting of the teeth on the jaw is also affected. All these make the face to look negative. So, to prevent orthodontic problem in children, it is necessary to detect the nasopharyngeal obstruction and treat the same accordingly. PMID- 18405093 TI - Rathke's cleft cyst--an uncommon cause of headache in a young boy. AB - Developmental disorders of the pituitary can present like a mass lesion in the region of the pituitary and produce pressure effects. Rathke's cleft cyst is one such developmental disorder of the pituitary which presents in early life producing pressure effects namely visual disturbances due to its proximity to optic chiasma, headache, and hormonal imbalance due to pituitary malfunction. In this case report a 19 years boy presented with symptoms of headache and gradual loss of vision over one year period. Neuro-imaging study demonstrated a cystic lesion in the region of the pituitary. Trans-sphenoidal hypophysectomy was performed. Histopathology of the biopsied material was suggestive of Rathke's cleft cyst. Following surgery the boy had a good visual recovery. So early diagnosis and removal of a Rathke's cleft cyst has a good prognosis as compared to a craniopharyngioma which is a very close differential diagnosis. PMID- 18405092 TI - Spontaneous cholecystocutaneous fistula in a case of chronic colculous cholecystitis--a case report. AB - Chronic calculous cholecystitis is quite a common disease entity today. However, advances in diagnostic imaging and use of broad-spectrum antibiotics have facilitated its early detection and treatment. Hence complications of untreated calculous cholecystitis have shown a steady decline. Spontaneous cholecystocutaneous fistula is one such rare complication. A 45-year-old female presented with acute onset of pain and swelling in the right hypochondrium. She had leucocytosis. The swelling ruptured spontaneously and 200 ml frank yellow pus along with multiple stones were expelled from the site. Completion drainage was done. A fistulogram confirmed the diagnosis of a cholecystocutaneous fistula. The patient underwent open cholecystectomy and excision of the fistulous tract. PMID- 18405094 TI - A rare case of fibrocystic disease at vulval accessory breast tissue. AB - A 40-year-old female presented with a non-itchy ulcerative nodular lesion at left labium majus since last 1 1/2 years. The lesion progressed to increase in size from 0.5 cm to 1.5 cm in diameter. It was incised and drained. After that a non healing ulcerative nodule formed. The nodule was firm in consistency and movable on all sides. The ulcer healed with a 5 days course of ceftriaxone. If was excised and biopsy of the lesion showed fibrocystic changes of accessory breast tissue. It is a rare disease entity for which the case report is presented. PMID- 18405095 TI - Torsion of appendices epiploicae of the colon: a case report. PMID- 18405097 TI - Quackery--a feedback discussion. PMID- 18405096 TI - Snakebite management in India. PMID- 18405098 TI - Effects of antacid therapy in asymptomatic patients with presence of amoeba in stool report. AB - Amoebic infection caused by the protozoan parasite Entamoeba histolytica is a prevalent infection in the developing countries. Milder form of this infection is associated with loose stool, flatulence and borborygmi, may or may not be associated with pain in abdomen and treated symptomatically by some physicians by antacid. To find out the effects of antacid (sorbacid) therapy in patients with amoeba in stools by examining the changes in the stool report, a study was conducted among 25 patients enrolled in the study with complaints of "gas" in the abdomen with stool reports positive for amoeba. Antacid (sorbacid) in a dose of one teaspoonful (5 ml) was given three times a day for 3 days and stool examination was repeated. The report showed a significant reduction in the amoeba and mucus in stool (p<0.05) and a trend towards reduction in the presence of occult blood. Other parameters in stool reports did not change. Moreover, all the patients gave the history of passing formed stools and no complaints of "gas" in abdomen thus providing the symptomatic benefit. Antacids may have some beneficial effects in amoebiasis. More studies are required to confirm the above finding and to find out the place of antacid as an adjuvant therapy along with the standard anti-amoebic drugs. PMID- 18405099 TI - Mass scans from a proton transfer mass spectrometry analysis of air over Mediterranean shrubland browsed by horses. AB - Plants usually emit large amount and varieties of volatiles after being damaged by herbivores. However, analytical methods for measuring herbivore-induced volatiles do not normally monitor the whole range of volatiles and the response to large herbivores such as large mammals is much less studied than the response to other herbivores such as insects. In this paper we present the results of using a highly sensitive proton transfer reaction-mass spectrometry (PTR-MS) technique that allows simultaneous monitoring of leaf volatiles in the pptv range. The resulting mass scans in air over Mediterranean shrubland browsed by horses show 70 to 100% higher concentrations of the masses corresponding to mass fragments 57, 43 and 41 (mostly hexenals, acetone and acetic acid) than scans over control non-browsed shrubland. These compounds are biogeochemically active and they are significant components of the volatile organic carbon found in the atmosphere. They influence the performance of living organisms and, the chemical and physical processes of Earth's atmosphere. PMID- 18405100 TI - Cytochrome P450 1A activity in liver and fixed wavelength fluorescence detection of polycyclic aromatic hydrocarbons in the bile of tongue-fish (Cynoglossus acrolepidotus, Bleeker) in relation to petroleum hydrocarbons in the eastern Gulf of Thailand. AB - This investigation was conducted in an area of oil spill along the east coast of Thailand to examine the relations among cytochrome P450 1A activity in liver and PAHs in the bile of the tonguefish and petroleum hydrocarbons in the sediments. PAH sediment concentrations in the reference and oil spill areas were 5.03 +/- 0.42 and 0.21 +/- 0.043 microg(-1) dry weight respectively Cytochrome activity in fish liver from oil spill area was 45.40 +/- 3.50 pmoles/ min/mg protein, almost threefold higher than that from the reference sites. Flourescense detection in bile metabolites at the oil spill area, 69.8 +/- 9.9 flourescense unit was significantly higher than that at the reference sites, 22.9 +/- 5.5 and 22.2 +/- 3.5 fluorescence unit. A strong correlation was found among cytochrome P450 1A activity in liver, PAH of bile metabolites and petroleum hydrocarbons. Both cytochrome and bile metabolites activity decreased seaward varying to the distance from the oil polluted area. We concluded that both detections in tonguefish can be regarded as a complementary biomarkers for the exposure of PAHs in tropical marine environments. PMID- 18405101 TI - Review on DDT and its residues in Turkey's wetlands. AB - Turkey is a country rich in wetlands (250 wetlands, approximately one million hectares). As it is in many countries, the wetlands in Turkey are threatened by the agricultural activities, loss of habitats due to these activities and by pesticide contamination. After 1940, large quantities of pesticides, especially DDT were used intensively against mosquitoes and pests in most of the wetlands in Turkey causing considerable contamination. In the beginning of 1980's, production, import and use of DDT was forbidden due to its toxicity on non-target organisms, environmental persistence, and accumulation in food chains. The scientific studies made in Turkey on the level of contamination in various environs and organisms by DDTand its metabolites, are limited These limited studies, especially the ones made after 1990's, have shown that we still observe DDT contamination in many wetlands (including internationally important ones like Goksu Delta and Meric Delta) and in organisms like birds and fishes. The data on the concentrations of DDTand its metabolites in the organisms and the biological magnification levels are not adequate since in most of the studies only water and sediment samples were analyzed. Besides, in the few studies made on the organisms, standard methods and suitable indicator organisms were not used. Because of these reasons, it is very difficult to make reliable evaluations on the DDT levels and its effects in the wetlands of Turkey. It is interesting that DDTlevels are higher than its metabolites in water sediment and organism samples in some wetlands which imply the possible use of DDT recently The aim of this review is to summarize the studies made so far on DDT and its residues in the wetlands of Turkey and to shed light on the possible hazardous effects and to propose approaches for future studies. PMID- 18405102 TI - Disinfection of wastewater: comparative evaluation of chlorination and DHS biotower. AB - The present study reports the onsite evaluation of two pilot scale disinfection units. One of the pilot plants is based on chlorination, and other is based on fixed film aerobic process (biotower). Evaluation study consisted of onsite monitoring of COD, BOD5 and TSS and fecal coliform over a period of three months. Samples were collected from the inlet and outlet of the pilot plants. These pilot plants were evaluated so as to have an appropriate disinfection technology for the treatment of the effluents from upflow anaerobic sludge blanket reactor (UASBR) based sewage treatment plants which could meet the biological quality standards. All the influents samples collected from both the pilot plants contained fecal coliform ranging from 105 to 10(6) MPN/100 ml. The results show that the fecal coliform removal is up to 98.2% and 100% forbiotower and chlorination, respectively. Both, the chlorination and down hanging sponge biotower (DHS-biotower) improved the quality of effluent from the UASBRin terms of COD, BOD5 and TSS. Though chlorination performed better compared to the DHS biotower, however, it has additional risk associated with the formation of trihalomethanes (THMs). PMID- 18405103 TI - Hawk moths (Lepidoptera: Sphingidae) of Turkey and their zoogeographical distribution. AB - The family Sphingidae (Lepidoptera) has 63 species in the western Palaearctic Region of the world. Thirty-four out of 63 species present in Turkey either permanently or temporarily. The subfamilies Smerinthinae, Sphinginae and Macroglossinae are consisted of 7, 4, and 23 species respectively Ten out of 34 species were captured in the field. Available knowledge of Sphingidae of Turkey was evaluated and summarized with this study as well. PMID- 18405104 TI - Comparison of chemical composition of the essential oil of Laurus nobilis L. leaves and fruits from different regions of Hatay, Turkey. AB - The essential oils of the leaves and fruits from bay (Laurus nobilis L.) grown in Antakya, Yayladagi and Samandagi were isolated by solvent extraction and analysed by capillary gas chromatography (GC), gas chromatography and mass spectrometry (GC/MS). In Antakya, Yayladagi and Samandagi the chemical compositions of the fruits and leaves were similar according to qualitative and quantitative analysis. Although in both fruits and leaves the major component was found to be 1.8-Cineole a concentration of about 50% compared with essential oils. The composition of the essential oil from the leaves has high content of 1.8-Cineole, Sabinene and alpha-Terpinyl acetate, but a low content of a-Pinene, alpha Phellandrene and trans-/beta-osimen. 1.8-Cineole was found major component of the leaves essential oil collected from Samandagi (59.94%) which is sea coast of region. Interestingly alpha-Pinene, beta-Pinene, alpha-Phellandrene, 1.8-Cineole and trans-beta-osimen were found the major components of fruits of Laurus nobilis L. harvested from Antakya, Yayladagi and Samandagi Trans-beta-osimen was detected as the major component of fruits essential oil collected again from Samandagi (28.35%) PMID- 18405105 TI - Biophoton emission of MDCK cell with hydrogen peroxide and 60 Hz AC magnetic field. AB - We studied biophoton characteristics of Madin-Darby canine kidney (MDCK) cells under the influence of H2O2 by employing a photomultiplier tube (PMT) and a fluorescence microscope. H2O2 was used for producing reactive oxygen species (ROS) in the measurement. Images from a fluorescence microscope show an increase of photon intensity emitted from the sample due to H2O2. By using a PMT we measured quantitative change in biophoton emission with application of H2O2 to the MDCK cell culture, found that the increase of the biophoton is dependent upon the amount of H2O2. The agreement between the results of the PMT and the fluorescence microscope suggests the possibility of quantitative measurement of the influence of ROS on living tissue or cell. In addition we applied a 60 HzAC magnetic field on the cells to investigate the change in reaction between MDCK cell and ROS. It showed that a decay of chemiluminescence intensity has taken a different path following exposure to the magnetic field. As a result, the PMT measurement might be considered as a useful tool for studying biochemical characteristics in relation to ROS. PMID- 18405106 TI - Solid state biomethanation of fruit wastes. AB - Overtones of fruit wastes accumulate daily in the city of Chennai, India and there is an urgent need to develop, assess and use ecofriendly methods to dispose them. Presently an attempt has been made to study solid state biomethanation of fruit wastes using a laboratory scale anaerobic digester Fruit wastes containing 3%, 4% and 5% solids were used for experimentation. Daily and cumulative production of biogas produced during the study was recorded. Increase in nitrogen, phosphorus and potassium content in the feed was observed during the digestion. During the study the biogas generation increased with increase in total solids. While the gas generation was 0.006 m3/day/m3 of the reactor volume when the solid content was 3% the corresponding values for 4% and 5% solids were 0.27and 0.35 m3 respectively Varying TS 3% to 4% has no effect on fermentation stability and pH remained between 6.8 and 7.4, but an inhibition of methanogenic bacteria was observed for TS 5%. The overall performance of the reactor was depressed by changing feed concentration from 3% to 5%. Experiments with 4% initial solid contend was ideal for solid-state biomethanation. PMID- 18405107 TI - Population dynamics of vibrio species in the river Narmada at Jabalpur. AB - Several studies on the presence and ecology of various Vibrio sp have been reported in coastal and estuarine waters throughout the world, but there is trifling information available on the distribution of this organism of colossal pathogenic potential in the fresh water riverine environment. Thus, we conducted a multiyearenvironmental study to scrutinize the occurrence of members of genus Vibrio in the largest west flowing river of the Indian subcontinent, which is also the largest river of central India, the Narmada. Statistical analysis was done to reveal major environmental factors controlling the presence of Vibrio sp in the river Narmada. Monthly field samplings were conducted between January 2002 and December 2003 at four different sites in Jabalpur (MP), India. At each site, water samples were taken and physicochemical and bacteriological parameters were measured. The identity of the isolates was confirmed by employing 16S rRNA analysis. The organisms were found to be widely distributed in the river with regular seasonal variations. The density of Vibrio was found to be correlated with temperature, coliforms and other heterotrophic bacteria. Water temperature accounted for most of the variability in the concentration of Vibrio spAs typical fecal pollution indicators may not access public health risk from potential pathogens such as vibrios, hence special monitoring programme for vibrios may adequately be included in the water quality management. PMID- 18405108 TI - Hepatoprotective effects of taurine against mercury induced toxicity in rats. AB - An attempt has been made to study the influence of taurine on mercury intoxicated rats. The animals were treated with sublethal dose of mercuric chloride (2 mg/kg body wt.) for 30 days. During the mercury treatment, the level ofAspartate transaminase(AST), Alanine transaminase (ALT) and Alkaline phosphatase(ALP) in serum and lipid peroxidation (LPO) in liver tissue significantly increased whereas Glutathione (GSH), Glutathione peroxidase(GPx), Catalase (CAT) and Superoxide dismutase (SOD) were simultaneously decreased in the liver tissue. Present results indicate that the liver tissue was completely damaged, after mercury treatment. In another group of animals, taurine (5 mg/kg body wt.) was administrated for another 15 days. Taurine administration was observed to improve the liver function in mercury intoxicated animal as indicated by the decline in increased levels of AST, ALT and ALP in serum and LPO content in liver tissue. The decreased level of antioxidant system (GSH, GPx, CATand SOD) has been promoted Results suggested that taurine played a vital role in reducing the mercury toxicity in intoxicated animals. PMID- 18405109 TI - Structure of immune organ in edible catfish, Mystus gulio. AB - The light microscopic study describes the anatomy and histomorphology of head kidney in bagrid catfish, Mystus gulio. Showing numerous lymphocytes, interrenal cells, reticular cells, postcardinal vein, blood sinuses and melanomacrophage centers. PMID- 18405110 TI - Cr (VI) and Fe (III) removal using Cajanus cajan husk. AB - Husk of tur dal (Cajanus cajan) was investigated as a new biosorbent for the removal of Fe (III) and Cr (VI) ions from aqueous solutions. Parameters like agitation time, adsorbent dosage and pH were studied at different initial Fe (III) and Cr (VI) concentrations. The biosorptive capacity of the Tur dal husk was dependent on the pH of the chromium and iron solution, with pH 2 and 2.5 respectively being optimal. The adsorption data fit well with Langmuir and Freundlich isotherm models. The practical limiting adsorption capacity (qmax) calculated from the Langmuir isotherm was 96.05 mg of Cr(VI)/ g of the biosorbent at an initial pH of 2.0 and 66.65 mg/g at pH 2.5. The infrared spectra of the biomass revealed that hydroxyl, carboxyl and amide bonds are involved in the uptake of Cr (VI) and Fe (III) ions. Characterisation of tur dal husk has revealed that it is an excellent material for treating wastewaters containing low concentration of metal ions. PMID- 18405111 TI - Effects of estradiol-17beta and 17alpha, 20beta-dihydroxy-4-pregnen-3-one on different phospholipids metabolism and histological changes in ovary during reproductive growth in the catfish, Heteropneustes fossilis (Bloch). AB - Effect of cumulative doses of estradiol -17beta (E2-7, 14 and 28 mg/kg body weight) and 17alpha, 20beta-dihydroxy-4-pregnen-3-one (17alpha,20betaP-7, 14 and 28 mg/kg body weight) on total phospholipids (TP) and various phospholipids- phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylinositol (PI) and phosphatidylethanolamine (PE) on liver plasma and ovary were investigated during the reproductively active preparatory and prespawning phases of the annual reproductive cycle in the freshwater female catfish, H. fossilis. The effect of E2 on TP was generally stimulatory and has pronounced effect than 17alpha,20betaP during both the phases. The levels of PC was promoted high during prespawning phase by E2 comparatively very less than by 17alpha, 20betaP in studied tissues during both the phases. The levels of PS after E2 treatments was maximum in all tissues during prespawning phase whereas 17alpha,20betaP was effective only in liver during this phase. The PI was elevated in liver during preparatory phase but its elevation was in all studied tissues during prespawning phase after E2 treatments. The levels of PI was most effective in ovary during preparatory phase in response to 17alpha,20betaP. The levels of PE was declined in liver but elevated in ovary after E2 treatments during both the phases. Treatments of E2 during preparatory phase showed greater number of vitellogenic oocytes as compared to 17alpha,20betaP treatments. The present finding has demonstrated that estradiol-17beta has more pronounced effects than the 17alpha,20beta P in regulation of different phospholipids and ovarian recrudescence during reproductively active phases and among the phospholipids the PC is the main phospholipids of vitellogenin/ovarian lipids in H. fossilis. PMID- 18405112 TI - Environmental impact of nitrogen fertilization in tea eco-system. AB - To quantify the nitrogen losses through runoff and leaching under a tea plantation in hilly soil, a field experiment was conducted from October 2001 to October 2002 at United Planters Association of Southern India (UPASI), Coonoorin Nilgiri district. Runoff water was collected in the collection tub on most rainy days but the leachate was collected in the soil water sampler when the rainfall exceeded 150 mm. Higher nitrogen fertilization levels significantly influenced the NO3-N concentration in both the runoff and leachate and it was likely to cause adverse environmental impact at the delivery end. The NH4-N and NO3-N concentrations in runoff decreased with the days after fertilizer application. NH4-N concentration reduced from 10.27 mg/l on the 9th day to 1.72 mg/l on the 34th day after fertilizer application. NO3-N concentration reduced from 23.5 mg/l on the 9th day to 4.32 mg/l on the 34th day after fertilizer application. Nitrogen loss varied depending on the quantity of rainfall and runoff. The NO3-N concentration in the leachate increased with increase in depth (18.06 mg/l at 22.5 cm depth to 20.98 mg/l at 45 cm depth) whereas NH4-N concentration decreased with increase in depth (6.32 mg/l at 22.5 cm depth to 5.79 mg/l at 45 cm depth. PMID- 18405113 TI - Antimicrobial activity of tissue and associated bacteria from benthic sea anemone Stichodactyla haddoni against microbial pathogens. AB - Associated bacteria from Stichodactyla haddoni are found maximum in tentacle tissues than the body tissue. There are eight associated bacterial species viz., Alcaligenes sp, Corynebacterium sp, Aeromonas sp, Sporosarcina sp, Renibacterium sp, Camobacterium sp1, Camobacterium sp2 and Salinococcus sp were recorded. The culture extracts from the associated bacterial species showed sensitivity against human bacterial and fungalpathogens. However, the hexane tissue extract of sea anemone showed maximum sensitivity (24 mm dia.) against the fish bacterial pathogen Aeromonas hydrophila than the other chosen pathogens. Comparatively the tissue extracts showed promising antimicrobial sensitivity than the cell free extracts of associated bacteria, and hence, the tissue samples from the sea anemone Stichodactyla haddoni is recommended for further exploration of novel antimicrobial drugs than the associated bacteria. PMID- 18405114 TI - Phosphate solubilizing actinomycetes in the estuarine environment: an inventory. AB - Sediment samples were collected from different stations of the Vellar estuary for isolation of total actinomycetes and phosphate solubilizing actinomycetes. Phosphatase activity in the sediments was also investigated Consistently a higher number of actinomycetes, phosphate solubilizing actinomycetes and phosphatase activity were recorded from the clay sediments than the sandy sediments at all the stations. In all, 7 strains showed positive phosphatase activity. Among them, one strain PS-3 exhibited good activity and was further investigated for optimum phosphorus solubilization at different pH (6, 6.5, 7, 7.5 and 8) and incubation (1st day to 20th day) periods. The solubilizing activity was maximum at the pH 7 and an incubation period of 13 days was required for an appreciable quantity of phosphorus to be leached into the medium. Based on the chemotaxonomical and conventional methods of identification, the strain PS-3 has been tentatively identified as Streptomyces galbus. The present study indicates that phosphatase enzyme and S. galbus along with other actinomycetes species would play a major role in solubilizing the phosphate in the estuarine ecosystem and increasing the soluble phosphate concentration thereby enhancing the productivity PMID- 18405115 TI - A comparative hydrobiological study of a few ponds of Barak Valley, Assam and their role as sustainable water resources. AB - A hydrobiological study conducted in nine different ponds of a rural area of Barak Valley, Assam, showed that the concentrations of chemical parameters like dissolved oxygen, free carbon dioxide, pH, conductivity alkalinity nitrate, phosphate, calcium, magnesium, copper and zinc are within the permissible levels of drinking water quality standard of WHO and ISI. However iron content was higher in most of the ponds. A clear indirect relationship between iron concentration and euglenoids has been observed. Major phytoplankton taxa present in the ponds are Chlorophyceae, Cyanophyceae, Bacillariophyceae and Euglenophyceae. The study reveals that rural ponds can be a very good source of water for drinking, domestic use and fishery and should be conserved at any cost. PMID- 18405116 TI - Life cycle of Lampito mauritii (Kinberg) in comparison with Eudrilus eugeniae (Kinberg) cultured on different substrates. AB - Growth (length, biomass and mean growth rate) and reproduction (total duration, clitellum appearance, clitellum completion, cocoon commencement, rate of cocoon production, incubation period, hatching success and mean number of hatching per cocoon) of indigenous Lampito mauritii (Kinberg) in comparison with exotic Eudrilus eugeniae (Kinberg) cultured on three feed substrates-clay loam soil, cowdung and pressmud (filter cake) have been studied over a period of 360 days under laboratory conditions (30 +/- 2 degrees C, 60-65% moisture). There is a positive relationship between length and biomass of both worms cultured on three feed substrates throughout the period of study The decrease of worm length and biomass observed slightly on 63-70th days in Lampito mauritii and 42-49th days in Eudrilus eugeniae cultured on three fed substrates are the results of the onset of cocoon production. After 270 days both worms in all these fed substrates show decreasing trends of length and biomass which are due to continued reproduction and aging. Among the three fed substrates, pressmud supports significantly maximum worm length and biomass (between 90-130 days in Eudrilus eugeniae and 110 170 days in Lampito mauritii), earlier attainment of sexual maturity (between 51 76 days in Limpito mauritii and 27-37 days in Eudrilus eugeniae), earlier commencement of cocoon production (37.7 +/- 0.0 days in Eudrilus eugeniae and 76.4 +/- 0.10 days in Limpitomauritii), shorter incubation periods (16.3 +/- 0.28 days in Eudrilus eugeniae and 26.7 +/- 0.81 days in Limpito mauritii), more hatching success (98% in Limpito mauritii and 86% in Eudrilus eugeniae), more mean number of hatchling percocoon (3.2 + 0.03 in Limpito mauritii and 2.6 +/- 0.06 in Eudrilus eugeniae) and shorter duration of life cycle (108.8 +/- 0.07 days in Limpito mauritii and 60.2 +/- 0.09 days in Eudrilus eugeniae) than cowdung and clay loam soil. PMID- 18405117 TI - Decolorization of remazol yellow RR gran by white rot fungus Phanerochaete chrysosporium. AB - In this study, the removal of color, chemical oxygen demand (COD) and aromatic group from one of the azo dyes, Remazol Yellow RR Gran, had been carried out by using one of the white rot fungi, Phanerochaete chrysosporium. Experimental studies were performed in growth media containing different amounts of dye and glucose. Color measurements were done at 436nm wavelength using spectrophotometer while aromatic group measurements were done at 280 nm wavelength using UV/Visible spectrophotometer. As a result of this study the values of the removable color concentration were determined as 10 mgl(-1) and lower. The optimum medium glucose concentration was determined to be 2 gl(-1) during color removal processes, aromatic group measurements were done in samples in the UV region at 280 nm wavelength. As a result of the measurements, it was shown that certain amount of aromatic group remained in the model wastewater at the end of the process. PMID- 18405118 TI - Isolation of Thiobacillus sp from aerobic sludge of distillery and dairy effluent treatment plants and its sulfide oxidation activity at different concentrations. AB - In the present study two strains of Thiobacillus sp were isolated from aerobic sludge of distillery and dairy effluent treatment plant using standard methods of isolation and enrichment. Experiments were conducted using isolated cultures in batch bioreactor with initial sulfide concentration of 75 and 150 mg/l. The effect of initial sulfide concentration on the activity of isolated Thiobacillus sp was studied. Sulfide oxidizing capacity was also determined at different initial sulfide concentrations. The results from the study indicate the possible isolation of Thiobacillus cultures from native source and application in the full scale reactor PMID- 18405119 TI - Acute toxicity effect of the venom of the marine snail, Conus zeylanicus. AB - Acute toxicity of the venom of Conus zeylanicus was studied to evaluate its risk and toxic factors in view of human safety The lethality of the crude venom (LD50 60 mg/kg via i.p.) in mice was associated with increased heart rate and strong muscular hind limb paralysis, skeletal muscle paralysis, dyspnea, loss of spontaneous activity followed by respiratory failure. The effect on vital tissues revealed liver tissues were disrupted with hemorrhagic necrosis and the lung showed the pathogenic changes of diffused inflammation of the parenchyma and obliteration of the alveolar space. In brain, edema was observed throughout the parenchyma and the kidney shows the tubules with cloudyswelling of the lining cells and the parenchyma inflammation and few inflammatory cells infiltration. PMID- 18405120 TI - Relative susceptibility of some common mosquito vector larvae to synthetic insecticidal compounds in north-western Rajasthan. AB - Relative susceptibility of three important mosquito vector larvae viz., Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, prevalent in the arid region was determined against four organophosphates (malathion, fenitrothion, fenthion, temephos) and three synthetic pyrethroid (alphamethrin, deltamethrin and fanvalerate) compounds. Studies were carried out on late 3rd or early 4th instar larvae of these species using standard WHO technique. Based on concentration mortality data LC50 and LC90 values along with their fiducial limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined by log probit regression analysis. LC50 values as observed for the above seven insecticides were 0.8097, 0.0398. 0.0432, 0.0035, 0.0025, 0.0092, 0.1006; 1.2370, 0.0531, 0.0655, 0.0076, 0.00004, 0.00004, 0.0046 and 1.4980, 0.0719, 0.0817, 0.0056, 0.00021, 0.00073, 0.0112 mg/l for the above three mosquito species respectively Among the four organophosphates tested temephos was the most effective followed by fenitrothion, fenthion and malathion. In general, Anopheles was found more susceptible as compared to the other two culicines to the above four compounds. The results also showed that larvae of Ae. aegypti were most susceptible followed by Cx. quinquefasciatus and An. stephensi to all the three pyrethroids tested. Among the three compounds tested alphamethrin was found to be the most toxic followed by deltamethrin while fanvalerate was the least toxic. The study would be of great importance while planning use of these insecticides for the control of different vector species in this area. PMID- 18405121 TI - Antihyperglycemic and antihyperlipidemic effects of Tephrosia purpurea leaf extract in streptozotocin induced diabetic rats. AB - Diabetes mellitus is a worldwide leading metabolic syndrome, associated with profound alterations in carbohydrate, lipids, lipoproteins and protein metabolisms. Worldwide, traditional practitioners for the treatment of diabetes and its complications use a wide variety of medicinal plants. In the present study the aqueous extract of Tephrosia purpurea leaves (TpALet) was evaluated for its antihyperglycemic and antihyperlipidemic effects in streptozotocin induced diabetic rats. Profound alterations in the concentrations of blood glucose, lipids and lipoproteins were observed in diabetic rats. Oral administration of TpALet to diabetic rats at a dose of 600 mg/kg body weight significantly reduced the level of blood glucose and increased the level of plasma insulin as well as normalized the lipids and lipoproteins profile. The present study thus demonstrated that TpALet has prominent antihyperglycemic and antihyperlipidemic effects in streptozotocin induced diabetic rats. PMID- 18405122 TI - Dust exposures in tractor and combine operations in eastern Mediterranean, Turkey. AB - The objective of this study was to determine the dust concentrations in selected tractor and combine operations in eastern Mediterranean region in Turkey Mean dust concentrations were 137.9 mg m(-3), 83.6 mg m(-3), 80.3 mg m(-3), and 88.8 mg m(-3) respectively for soil packing, furrowing, straw making, and baling on tractors with no cabins whereas 106.9 mg m(-3) was found in combines without cabins, which are much higher than the limit dust concentration (10 mg m(-3)) considered hazardous for workers' health. In tractor operations with field installed cabins, mean dust concentrations were 5.6 mg m(-3), 6.6 mg m(-3), 6.4 mg m(-3), and 3.7 mg m(-3), respectively in soil packing, furrowing, straw making, and baling while 4.7 mg m(-3) was measured in combines with field installed cabins. Considering unit-manufactured cabins, mean dust concentrations were 1.1 mg m(-3), 1.6 mg m(-3), 3.2 mg m(-3), and 1.4 mg m(-3) respectively in tractor operations, and 1.4 mg mr(-3) in combine operations. Variance analyses showed that not only the respiration environment of the workers but also the field operation had a significant effect on measured dust concentrations (p < 0.01). Pearson correlation coefficient was 0.74 (very strong level) between dust concentration and wind speed in soil packing, 0.46 (medium level) between dust concentration and wind speed in baling, and 0.44 (medium level) between dust concentration and ground speed in combining. PMID- 18405123 TI - Concentration of heavy metals in water and chub, Leuciscus cephalus (Linn.) from the river Yildiz, Turkey. AB - The concentration of heavy metals (Cu, Fe and Zn) were determined by atomic absorption spectrophotometry in the river water and the edible muscle tissues of chub, Leuciscus cephalus, from river Yildiz, Turkey, and in the waste water The following results were found in the water of the river Yildiz: Cu 0.03-0.53, Fe 0.91-1.96 and Zn, 053-1.49 microgl(-1), in the waste water Cu 0.20-0.52, Fe 1.22 2.29 and Zn 0.92-1.46 microgl(-1) and in the edible muscle of chub: Cu 1.00-3.79, Fe 7.21-17.04 and Zn 4.12-18.33 microg g(-1) wet weight respectively. Among the heavy metals studied Pb, Cd, Co, Cr, Ni and Mn were not detected in the river water, waste water and chub samples. Heavy metal contents in these samples were evaluated and the highest concentrations of Cu, Fe and Zn were found in the muscle tissue. The levels of the heavy metals were detected in decreasing order as iron> zinc> copper. All the samples contained comparatively lower amounts of metals as suggested by international and national regulatory bodies. Thus, we recommend periodic monitoring of these metals in the fish consumed by local people. PMID- 18405124 TI - Quantification of transition metals in biological samples and its possible impact on ferro-alloy workers. AB - Increased risk of ill-health and diseases has been associated with employment in the ferro-alloy factory. Since measurement of transition metals in human blood and hair along with respective exposure rates, provides a means of assessing individual risk, it has been the most important part of the study. In the study majority of the elements in the transition series, such as, vanadium (V), chromium (Cr), iron (Fe), manganese (Mn), cobalt, (Co) nickel (Ni), copper (Cu), zinc (Zn), molybdenum (Mo) and cadmium (Cd) were considered which are randomly emitted from the source, that is, manganese ore (used during ferro-alloy manufacturing process). The commonly available transition, metals, observed in biological samples of ferro-alloy workers, were found to be Fe, Zn, Co, Ni, Cu, Cr, Cd, V Mn and Mo in blood, while in hair, Mn, Fe, Zn, Co, Ni, Cu, Cr, Cd, V and Mo were present in decreasing order Surveillance of bio-concentration of these metals in workers, exposed to close proximity of the coke-ovens and smelting furnaces, revealed that the workers were prone to several physical disorders. PMID- 18405125 TI - Evaluation of antifouling activity of eight commercially available organic chemicals against the early foulers marine bacteria and Ulva spores. AB - Environmental impacts caused by tin and copper based commercial antifouling (AF) paints were proved to be detrimental to aquatic ecosystems. Therefore, a search of environmental friendly AF compounds to be used in marine paint to protect the surface of maritime developmental structures from the unwanted biofouling is a burning issue of the present time. Commercially available eight organic chemicals -allyl isothiocyanate, beta-myrecene, cis-3-hexenyl acetate, citral, ethyl heptanoate, eugenol, methyl caproate, and octyl alcohol were evaluated forAF activities using both laboratory and field assays. The test chemicals were found to repel the target motile marine bacteria--Alteromonas marina, Bacillus atrophaeus, Roseobactergallaeciensis and Shewanella oneidensis and motile spores of the green alga, Ulva pertusa. The bacterial and Ulva spore repulsion activities of the test chemicals were measured by chemotaxis and agar diffusion methods respectively interestingly these test chemicals were less toxic to the test fouling species. The toxicity of the test chemicals was measured by using antibiotic assay disks against the bacteria and motility test against Ulva spores. Moreover, in field assay, all test chemicals showed a perfect performance ofAF activity showing no fouling during the experimental period of one year Such results and commercial as well as technical feasibility of the test chemicals firmly showed the possibility of using as alternatives of the existing toxic AF agents. PMID- 18405126 TI - HIV sero-prevalance in general population of Warangal, A.P., South India. AB - HIV/AIDS pandemic has devastated many countries reversing national development; HIV was not seen in Asia and India till 1980. Now India has become epicenter of AIDS pandemic. During April 2002 to March 2003 the HIV+ ve pregnant women and their husbands were encouraged to enroll in the prospective study with informed consent. The study results consist of most of the females who are in the age group between 16-25 years who were affected by HIV High infection is observed in people with lower socio-economic and education background. High infection rate is observed in house wives (26.7%), laborers (23%) and agricultural workers (12.1%) followed by toddy tapers (5%), drivers (5.96%) and others (6 47%). HIV +ve subjects at Mother To Child Transmission (MTCT) centers are surprisingly clinically very healthy. No disease manifestation was noticed. PMID- 18405127 TI - Pre-diabetes, dysglycaemia and early glucose intolerance and vascular health. PMID- 18405128 TI - Risk of future diabetes is as high with abnormal intermediate post-glucose response as with impaired glucose tolerance. AB - AIMS: This analysis was done to compare the risk of development of diabetes among subjects with impaired glucose tolerance (IGT) and early glucose intolerance (EGI; intermediary post glucose level > or = 160 mg/dl) when compared with normal (NGT). Profile of insulin secretion and insulin resistance was compared in a subgroup of subjects with EGI, IGT and diabetes. PATIENTS AND METHODS: A program on 'primary prevention of diabetes' was initiated and high risk subjects were encouraged to join the program and were followed up. Out of 4084 (M:F 2344:1740) subjects enrolled in the program, a total of 1659 (M:F 1044:615) subjects with mean age 41.3 +/- 10.2 years who had at least two follow up visits were selected for this analysis. OGTTs were performed once in every 6 months. The median follow up duration was 5 years. The conversion rate to diabetes in subjects with persistent IGT or EGI was determined. In a subgroup of subjects, NGT (n = 118), IGT (n = 68), EGI (n = 106) and new DM (n = 126), plasma insulin at fasting, 30 min and 2 hr were measured and insulin resistance (HOMA-IR) was calculated by HOMA method. Insulinogenic index (DeltaI/G) was also calculated. RESULTS: The rate of conversion of IGT 251 (40.5%) and EGI 210 (36.5%) subjects to diabetes was similar and significantly higher when compared with the NGT subjects 99 (21.3%). Similar results were noted both in men and women. By using ROC procedure, a cut - off value of one hour post glucose of > or = 160 mg/dl gave a sensitivity of 80% and specificity of 70% to detect abnormal glucose tolerance. In a subset of subjects studied, it was noted that subjects with EGI had significantly higher fasting insulin values than NGT. Insulin resistance (IR) was the highest in DM [Geometric mean (SD)] 6.6 (1.9)), followed by EGI (4.5 (2.3)) (p < 0.0001 vs NGT, 2.9 (2.4)) and IGT (3.9 (2.2)). Insulinogenic index was normal in EGI, NGT and IGT, and it was lower in DM in comparison with other groups. The multiple logistic regression analysis showed that EGI (odds ratio (OR) 2.11) and development of diabetes was strongly associated. The survival curve (time free from diabetes) showed that the median survival time for NGT, EGI and IGT were 18.7, 11.6 and 9.6 yrs respectively. CONCLUSION: EGI which is a distinct entity with abnormal intermediate glucose response in glucose tolerance test (GTT) appears to be a risk factor similar to IGT in the development of diabetes. They had higher insulin resistance with normal insulin secretion. Therefore, it is important to determine the intermediate value also during the GTT in addition to fasting and 2 hr post glucose values. PMID- 18405129 TI - Pulmonary manifestations in systemic lupus erythematosus (SLE) with special reference to HR CT. AB - AIMS AND OBJECTIVES: 1. To evaluate whether CT helps in the early diagnosis of pulmonary manifestations in SLE. 2. To study the pulmonary involvement and determine the incidence and type of pulmonary involvement. MATERIALS AND METHODS: Thirty eight patients fulfilling ACR classification criteria for SLE were studied. Chest X-ray, pulmonary function tests (PFT), HRCT scan thorax were done. Investigations were done to detect other organ involvement. Patients with known interstitial lung disease (occupational hazard) or pregnant females were excluded. RESULTS: Clinical signs and symptoms referable to pulmonary involvement were present in 9 patients (23.68%). HRCT thorax detected abnormalities in 21 patients (55.26%); pulmonary function abnormalities were present in 11 patients (28.95%) and plain X-ray chest abnormalities were present in 7 patients (18.42%). Out of 38 patients pulmonary involvement was present in 22 (57.89%) cases and HRCT thorax findings were present in 21 (95.45%) of them. CONCLUSION: Pulmonary involvement is present in a significant number of SLE patients, who are often asymptomatic, with normal chest X-ray or normal PFT. HRCT scan thorax could diagnose the pulmonary involvement in asymptomatic patients with normal chest X ray and uncertain PFT. PMID- 18405130 TI - Outcome of sixty four cases of ethylene dibromide ingestion treated in tertiary care hospital. AB - BACKGROUND AND AIMS: In view of high incidence and mortality due to ethylene dibromide poisoning, a prospective study was planned to assess demographic, toxicological, clinical profile and outcome. MATERIAL AND METHODS: Prospectively collected data of sixty four patients with ethylene dibromide poisoning were reviewed. All the patients were from Gwalior and neighbouring districts. RESULTS: Majority were males in the age group 21-30 years. Out of sixty-four cases 26 patients (40.6%) survived and 38 patients (59.4%) expired. Nausea vomiting and pain abdomen were most common symptoms at presentation. Diarrhoea, drowsiness, palpitations and oliguria were other features. Pulse rate was normal (< 100) in 22 (84.6%) survived patients whereas it was > 100 in 24 (63.1%) of expired patients at the time of admission (p value = 0.0004199). Half ampoule (1.5ml.) of ethylene dibromide was sufficient as fatal dose. Mortality was 20% and 46.15% with half and one ampoule respectively. Best outcome was when the patient presented to the hospital within six hours. Death occurred anywhere between twelve hours and five days. Gastrointestinal toxicity was predominant at presentation. Nausea, vomiting and abdominal pain was present in all the patients. Nephrotoxicity 32 (50%), hepatotoxicity 28 (43.8%), cardiotoxicity 28 (43.8%), central nervous system 8 (12.5%) toxicity and hypoglycemia 24 (37.5%) were also observed. CONCLUSION: Ethylene dibromide is highly toxic. Patients who consumed half to one ampoule usually survived with supportive care provided they presented to the hospital within six hours. Pulse rate more than 100 at presentation was a poor prognostic factor. Patients with normal pulse and blood pressure at admission had better prognosis. PMID- 18405131 TI - Etiopathogenesis and predictors of in-hospital morbidity and mortality in posterior circulation strokes--a 2 year registry with concordant comparison with anterior circulation strokes. AB - AIMS: To determine etiopathogenetic factors, predictors of in-hospital morbidity and mortality, and discharge status in a serially recruited cohort of PCS patients at a tertiary care hospital. Comparison of data between ACS and PCS groups was also done. METHODS: Seventy six cases of PCS and 108 cases of ACS were serially recruited, within 48 hrs of stroke onset, over a 2 year period. Vascular territory determination, stroke subtype and classification, risk factor profile and outcome measures were determined. RESULTS: 77.6% of PCS strokes were ischemic in origin. A cardioembolic source was seen in 12/29 (42%) PCS cases. Intraarterial cause was seen in 5/29 (17.2%) PCS cases. 22.3% of PCS and 24% of ACS patients developed dysphagia. Pneumonia developed in 70% and 65% of dysphagic patients in PCS and ACS groups respectively. Mortality in PCS group was 14/76 (18%) and in ACS group 17/108 (15.7%). The principal contributory factors to mortality in PCS were low Glasgow coma score at presentation, development of respiratory morbidity, and vascular lesions in 'middle plus distal' territory. At discharge, 62% PCS patients were in group 2-3 of modified Rankin scale and 64% of ACS patients were in group 3-4. CONCLUSIONS: A cardiac source of distal territory infarct was significantly commoner in PCS as compared to ACS. Incidence of post stroke complications, viz. dysphagia, pneumonia, deep venous thrombosis, bed sores and urinary infection, was comparable in the two groups. Dysphagia, seen in 22% PCS and 24% of ACS, was the principal causation of pneumonia. Mortality and disability status at discharge were comparable in the 2 groups. PMID- 18405132 TI - Syringomyelia with syringobulbia. PMID- 18405133 TI - 'Himalayan' p wave. PMID- 18405134 TI - Helicobacter pylori link to pernicious anaemia. AB - An immunological classification of chronic gastritis based on the detection of Helicobacter pylori (H. pylori) antibody, parietal cell antibody, intrinsic factor antibody, is reported. H. pylori chronic gastritis, slowly progresses to atrophic gastritis, in the majority of patients; in a few patients, with genetic susceptibility to form intrinsic factor antibody, it progresses to pernicious anaemia. In majority of patients of pernicious anaemia, H. pylori gradually disappears from the gastric mucosa, on development of intestinal metaplasia in them. Atrophic gastritis results from H. pylori or non H. pylori. H. pylori infection is diagnosed in the presence of H. pylori in the gastric mucosal biopsy and/or H. pylori antibody (IgG) in the serum. The presence of the genetic factor (intrinsic factor antibody) is essential for the diagnosis of pernicious aneamia. Pernicious anaemia patients without intrinsic factor antibody, should be correctly diagnosed as atrophic gastritis, in view of the absence of the genetic factor (intrinsic factor antibody) in them. PMID- 18405135 TI - Medical ablation of endobronchial mucormycosis with Amphotericin-B. AB - A 56 years male diabetic patient presented with recurrent left upper lobe pneumonia. Fiberoptic bronchoscopy revealed extraluminal compression of left main bronchus with an endobronchial mass obstructing the left upper lobe orifice. The lesion resembled bronchial adenoma. However histological examination revealed mucormycosis. Timely diagnosis followed by medical intervention with intravenous Amphotericin B, coupled with proper management of diabetes, ablated the tumor. Relevant literature on the subject is reviewed. PMID- 18405136 TI - Celiac disease and postcricoid carcinoma. AB - Malignant disease of oropharynx, esophagus and small intestine occurs in patients of celiac disease frequently. The role of gluten-free diet in prevention of malignancy is unclear. We report a case of postcricoid carcinoma occurring in the patient of celiac disease. PMID- 18405137 TI - Spotted fever group rickettsioses in Himachal Pradesh. AB - Rickettsiae are in many places of the world emerging or reemerging pathogens. The spotted fevers group (SFG) composes a large group of tick- and mite-borne zoonotic infections that are caused by closely related rickettsiae. The SFG rickettsiae of Southeast Asia are yet to be identified. Earlier reports have documented the endemicity of rickettsioses among adults in the Himalayan belt but no confirmed case of spotted fever have been reported from this region of India. We present two cases of SFG rickettsioses; from the northern hilly region of India that were confirmed using specific microimmunofluorescence assay. PMID- 18405138 TI - Hair dye poisoning. AB - Hair dye ingestion is an uncommon form of poisoning in the west, however, in some parts of the world such as East Africa and Indian Sub-continent it is not uncommon. The main component of hair dye causing toxicity is Paraphenylenediamine (PPD). This compound has been found to cause angioneurotic edema, rhabdomyolysis and renal failure. We present a case of hair dye poisoning who presented with respiratory distress due to laryngeal edema and later developed trismus and carpopedal spasm. This case report highlights the combined toxicities of sodium EDTA and PPD. PMID- 18405139 TI - Luc Montagnier--discoverer of HIV virus. Postal stamps issued on Luc Montagnier. PMID- 18405140 TI - Thrombolysis: the elusive El-Dorado. PMID- 18405141 TI - Disseminated abdominal hydatidosis. PMID- 18405142 TI - Pleural effusion of a dual etiology. PMID- 18405143 TI - Infectious causes of macrophage activation syndrome. PMID- 18405145 TI - Clinical and economic implications of incretin-based therapies. PMID- 18405144 TI - Incretin enhancers and the evolution of antihyperglycemic therapy in type 2 diabetes. PMID- 18405146 TI - [A study on differences of curative effects of acupuncture and nCPAP for treatment of OSAHS]. AB - OBJECTIVE: To compare curative effects of acupuncture and nasal continuous positive airway pressure (nCPAP) on obstructive sleep apnea-hypopnea syndrome(OSAHS), and to explore the characteristics and advantages of acupuncture in treatment of this disease. METHODS: Sixty-six cases of OSAHS were randomly divided into an acupuncture group (n = 44) treated by acupuncture at Lianquan (CV 23) and Tianrong (SI 17) etc., and a nCPAP group (n = 22) treated by nCPAP. Indexes of respiration, blood oxygen and sleep were monitored before, during and after treatment. RESULTS: Compared with those before treatment, no significant differences of these indexes were found in the acupuncture group during treatment (P > 0.05), but hypopnea index (HI), apnea-hypopnea index (AHI), the maximal apnea duration, the maximal hypopnea duration, sleep time spent when SaO2 was below 90% (SaO2 < 90%T) and the microarousal index improved significantly at the end of treatment (P < 0.05 or P < 0.01); while all the indexes improved significantly in the nCPAP group during treatment (P < 0. 01), but the therapeutic effect was not maintained at the end of treatment with no significant difference as compared with those before treatment. The therapeutic effect in the nCPAP group was better than that in the acupuncture group during the treatment (P < 0.05 or P < 0.01), but no significant difference of the therapeutic effect was found between the two groups at the end of treatment (P > 0.05). CONCLUSION: The therapeutic effect of nCPAP on OSAHS produces during treatment and the therapeutic effect of acupuncture produces after treatment, indicating they possibly have different mechanisms. PMID- 18405147 TI - [Randomized controlled study on acupuncture for treatment of diabetic paralytic squint]. AB - OBJECTIVE: To observe clinical therapeutic effect of acupuncture on diabetic paralytic squint. METHODS: Seventy-two cases of diabetic paralytic squint were randomly divided into a medication group, an acupuncture group and an acupuncture and medication group. The medication group were treated with intramuscular injection of Methyl vitamin B12 250 microg, once daily; the acupuncture group were treated by acupuncture at different acupoints according to different paralytic muscles of eyes with adjuvant acupoints selected according to symptoms; the acupuncture and medication group were treated with the routine medicine and acupuncture. The treatment was given for 28 days. RESULTS: The total effective rate of 87. 5% in the acupuncture group and 95.7% in the acupuncture and medication group were higher than 54.5% in the medication group (P < 0.05, P < 0.01), with no significant difference between the acupuncture group and the acupuncture and medication group (P > 0.05). CONCLUSION: Acupuncture has a definite therapeutic effect on diabetic paralytic squint, which is better than that of routine medication. PMID- 18405148 TI - [Clinical observation on treatment of primary knee osteoarthritis of liver and kidney deficiency type with Aconite cake-separated moxibustion]. AB - OBJECTIVE: To observe the effect of Aconite cake-separated moxibustion on primary knee osteoarthritis of liver and kidney deficiency type. METHODS: Fifty-six cases of such disease (80 knees) were randomly divided into a cake-separated moxibustion group (41 knees) with Neixiyan (EX-LE 5), Dubi (ST 35), Yinlingquan (SP 9) etc. selected, and a western medicine group (39 knees) were treated with oral administration of Sodium Diclofenate Slow-released Tablet. RESULTS: The cumulative score for symptoms and signs was (37.41 +/- 6.61) points before treatment and (9.37 +/- 8.15) points after treatment in the cake-separated moxibustion group and (37.64 - 6.00) points before treatment and (12.05 +/- 8.83) points after treatment in the western medicine group, with a very significant difference before and after treatment in the two groups (P < 0.01); two months after treatment, it was (11.71 +/- 8.69) points in the cake-separated moxibustion group and (15.95 +/- 9.96) points in the western medicine group, the former being better than the latter (P < 0.05). After the end of treatment, there was no significant difference in the comprehensive therapeutic effect between the two groups (P > 0.05), the cured and markedly effective rate was 63.4% in the cake separated moxibustion group and 48.7% in the western medicine group, but two months after treatment, the cured and markedly effective rate of 56.1% in the cake-separated moxibustion group was better than 33.3% in the western medicine group (P < 0.05). CONCLUSION: Aconite cake-separated moxibustion has an ideal therapeutic effect on primary knee osteoarthritis of liver and kidney deficiency type, and the therapeutic effect at anaphase is better than that of Sodium Diclofenate Slow-released Tablet. PMID- 18405149 TI - [Clinical study of acupuncture and moxibustion for preventing nosocomial infection in the patient with acute cerebrovascular disease]. AB - OBJECTIVE: To observe the effect of acupuncture and moxibustion for preventing nosocomial infection in the patient with acute cerebrovascular disease. METHODS: One hundred cases of acute cerebrovascular disease were randomly divided into a control group and an observation group, 50 cases in each group. The control group were treated by routine therapeutic method without any method for preventing nosocomial infection; the observation group were treated by the routine treatment, with acupuncture and moxibustion added at Guanyuan (GV 4) and Zusanli (ST 36), once daily, for 10 days. The nosocomial infection rate and the abnormal rates of body temperature, WBC and neutrophil were investigated in the two groups. RESULTS: The nosocomial infection rate was 8.0% in the observation group and 26.0% in the control group with a significant difference between the two groups (P < 0.05), and the abnormal rates of body temperature, WBC and neutrophil in the observation group were significantly lower than those in the control group (all P < 0.05). CONCLUSION: Acupuncture and moxibustion can effectively prevent nosocomial infection for the patient with acute cerebrovascular disease. The mechanism possibly is related with regulative action of acupuncture and moxibustion on immunity. PMID- 18405150 TI - [Effects of acupuncture on body mass index and waist-hip ratio in the patient of simple obesity]. AB - OBJECTIVE: To observe the therapeutic effect of acupuncture for slimming. METHODS: Eighty cases of simple obesity were randomly divided into an acupuncture group and a medication group, 40 cases in each group. The acupuncture group were treated with body acupuncture, electroacupuncture and auricular acupoint taping and pressing, with Tianshu (ST 25), Guanyuan (CV 4), Sanyinjiao (SP 6) etc. selected for body acupuncture, and auricular acupoints Shenmen, Neifenmi (endocrine), Pi (spleen) etc. selected for ear acupoint taping and pressing. And the medication group were treated with oral administration of Sibutramine. Body weight, body mass index, waist and hip circumferences and waist-hip ratio were determined before and after treatment to evaluate therapeutic effect of slimming. RESULTS: The total effective rate of 87.5% in the acupuncture group was similar to 82.5% in the medication group (P > 0.05). After treatment, body weight, body mass index, waist and hip circumferences and waist-hip ratio significantly decreased in the two groups, but the improvement of waist circumferences and waist-hip ratio in the acupuncture group was better than that in the medication group (P < 0. 05). CONCLUSION: Acupuncture has a definite therapeutic effect in slimming and it can effectively improve body weight, body mass index, waist and hip circumferences and waist-hip ratio in the patient of simple obesity. PMID- 18405151 TI - [Observation on therapeutic effect of comprehensive therapy for acute large area cerebral infarction]. AB - OBJECTIVE: To search for a method for increasing clinical therapeutic effect of acute large area cerebral infarction. METHODS: Ninety-six cases were randomly divided into a comprehensive treatment group and a western medicine group, 48 cases in each group. The western medicine group were treated with routine western methods, dehydration for decreasing intracranial pressure, brain-protection, anti platelet agglutination, improving circulation and expectant treatment, etc.; the comprehensive treatment group were treated with the routine treatment of western medicine combined with acupuncture, moxibustion and enema of Chinese drugs. The therapeutic effect was observed after treatment for 28 days. RESULTS: The total effective rate was 68.8% in the comprehensive treatment group and 39.6% in the western medicine group with a very significant difference between the two groups (P < 0.01), and the score of the National Institute of Health's Stroke Scale (NIHSS) and the score of the Glasgow Coma Scale (GCS) in the comprehensive treatment group were better than those in the western medicine group (P < 0.001, P < 0.01). CONCLUSION: Combined treatment of Chinese medicine and western medicine for large area cerebral infarction is better than simple western medicine, and it can significantly improve neurological defect of the patient and decrease death rate. PMID- 18405152 TI - [Analysis on therapeutic effect of selective acupoint-injection on scissors gait in children of cerebral palsy]. AB - OBJECTIVE: To observe the effect of selective acupoint-injection on scissors gait in 60 children of cerebral palsy. METHODS: Sixty children of scissors gait were classified hypermyotonia and hypomyotonia according to muscular tension and femoral angle, and treated with injection into lateral acupoints, medial acupoints and lateral-medial acupoints of the lower limbs, respectively. RESULTS: The total effective rate was 87.5% for the hypermyotonia treated with injection into lateral acupoints of the lower limbs, and 100.0% for the hypomyotonia treated with injection into the medial acupoints of the lower limbs, and 80.0% for the patient treated with lateral-medial acupoints. CONCLUSION: The causes of forming scissors gait for children of cerebral palsy should be analyzed and different acupoints should be selected for different causes, and different acupoints have different therapeutic effects. PMID- 18405153 TI - [Some misunderstandings of deqi phenomenon: from historic review to experimental study]. AB - In this paper, the authors trace back to the historic source of deqi and needling sensation, and probe the misunderstanding course of the feeling of sourness, numbness, distension, aching or heaviness used as synonym of deqi, and explore and introduce the modern significance and experimental studies of deqi. Although there are arguments in relationship between the feeling of sourness, numbness, distension, aching or heaviness after acupuncture and effect of acupuncture at home and abroad, in fact, this is a misunderstanding of needling sensation replacing deqi. Sometimes acupuncture possibly does not induce sourness, numbness, distension, aching or heaviness, but it also induces patient's comfortable sensation and at the same time acupuncture practitioners possibly have a sensation under the hand, which is at the state of deqi. The two states will be detected by fMRI in experimental studies, so as to have indexes to be followed for deqi or needling sensation. PMID- 18405154 TI - [Randomized controlled study on reinforcing method of acupuncture for treatment of Bell's palsy at restoration stage]. AB - OBJECTIVE: To observe clinical therapeutic effect of reinforcing method of acupuncture on Bell's palsy at restoration stage. METHODS: One hundred and sixty cases were randomly divided into two groups: an observation group and a control group, 80 cases in each group. The observation group were treated by acupuncture with lifting-thrusting-twirling reinforcing method, and the control group by acupuncture without needling manipulation. The therapeutic effect, and changes of palsy index, degree of palsy and ENoG were observed. RESULTS: Reinforcing method of acupuncture could significantly improve palsy index, degree of palsy and maximal wave amplitude of ENoG; the cured and markedly effective rate was 91.3% in the observation group and 75.0% in the control group with a very significant difference between the two groups (P < 0.01). CONCLUSION: The reinforcing method of acupuncture can increase the therapeutic effect on Bell's palsy at restoration stage. PMID- 18405155 TI - [Observation on therapeutic effect of moxibustion at "heat sensitive points" on perennial allergic rhinitis]. AB - OBJECTIVE: To compare therapeutic effects of moxibustion at "heat sensitive points" and western medicine on perennial allergic rhinitis. METHODS: One hundred and twenty cases were randomly divided into a moxibustion group and a western medicine group, 60 cases in each group. The moxibustion group were treated with moxibustion at "heat sensitive points" on the head-face, abdomen, waist and back, once a day; and the western medicine group with oral administration of Cetirizine Hydrochloride, 10 mg each session, once daily, 10 days constituting one course. RESULTS: The total effective rate and the total effective rate for the cases of over effectiveness 3 months later were 85.0% and 86.3% in the moxibustion group and 63.3% and 50.0% in the western medicine group, respec tively, with significant or very significant difference (P < 0.05, P < 0.01). CONCLUSION: Moxibustion at "heat sensitive points" is an ideal therapy for perennial allergic rhinitis with low recurrence rate. PMID- 18405156 TI - [Study on effects of acupuncture on mice dysmenorrhea model and the mechanism]. AB - OBJECTIVE: To probe the mechanism of acupuncture in treatment of dysmenorrhea. METHODS: Adult mice with no pregnancy were randomly divided into a normal group, a model group, an acupuncture group and a medication group. The model group, the acupuncture group and the medication group were modeled by Diethylstilbestrol and Ocytocin. For the acupuncture group, at the 7th day of modeling, acupuncture was given at "Sanyinjiao" (SP 6), "Diji" (SP 8), once a day, for 5 days; and at the 7th day of modeling, Yimucao Gao 0.6 mg/g was given intragastrically to the medication group for 5 days. The stretching latent period and the number of stretching within 30 min were observed, and mRNA levels of ocytocin receptor (OctR) and vasopressin receptor (VasR) in the uterus tissue were detected with RT PCR method. RESULTS: Compared with the model group, the stretching latent period extended (P < 0.05) and the number of stretching within 30 min significantly decreased (P < 0.05); and there were significant differences in the mRNA levels of ocytocin receptor and vasopressin receptor in the uterus tissue in the model group as compared with those in other 3 groups (P < 0.05, P < 0.01). CONCLUSION: Acupuncture can improve the dysmenorrheal symptom to a certain extent, and the mechanism is possibly related to regulative effects of acupuncture on hormones mediating receptors in mice. PMID- 18405157 TI - [Electroacupuncture alleviates complete Freund's adjuvant peripheral chronic inflammatory pain in mice]. AB - OBJECTIVE: To probe into the law of acupuncture analgesic effect and specificities of acupoint action. METHODS: Adult male Kunming mice were randomly divided into a control group, a model group, a model plus electroacupuncture (EA) group and a model plus sham EA group. Chronic inflammatory pain model was prepared by injection of complete Freund's adjuvant (CFA) into right posterior foot, and paw withdrawing latency (PWL) induced by radiation heat was used as pain threshold index, and changes of PWL in all the groups were investigated. RESULTS: After modeling, PWL significantly shortened on the inflammatory side (P < 0.05); EA at bilateral "Zusanli" (ST 36) and "Kunlun" (BL 60) could significantly reverse the shortened PWL (P < 0.05), and this effect was prolonged along with increase of EA times, but in the sham EA group PWL did not significantly change; EA at bilateral "Shousanli" (LI 10) and Neiguan" (PC 6) could not reverse the shortened PWL (P > 0.05). CONCLUSION: Analgesic effect of EA is strengthened along with increase of EA times and shows specificity of acupoints to a certain extent. PMID- 18405158 TI - [Chief physician SHAN Yong-Hua's clinical experiences]. AB - Chief physician SHAN Yong-Hua is engaged in clinical and teaching work for more than 40 years, with excellent medical techniques. He widely experiences variety of good methods and follows ancient theory, applies Jiehuo needling method in Miraculous Pivot, Cijie Zhenxie, and adopts "Kaiqiao Jiehuo " method, "Huayu Jiehuo" method, "Pingheng Jiehuo" method to treat syndromes at different stages of stroke. He is good at using scalp acupuncture for treatment of varieties of difficult and complicated diseases, and emphasize application of special acupoints in clinic practice. In process of acupuncture, he emphasizes needling manipulation methods, treating mind and keeping mind, promoting qi reaching to the affected area, hence increasing clinical therapeutic effect. PMID- 18405159 TI - [Acupuncture combined with auricular point sticking and pressing for smoking cessation of 53 cases in Russia]. AB - OBJECTIVE: To observe therapeutic effect of acupuncture combined with auricular point sticking and pressing for smoking cessation. METHODS: Body acupoints for acupuncture, Jieyan (two horizontal fingers above styloid process of radius), Shenmen (HT 7), Zhongwan (CV 12), Zusanli (ST 36), Sanyingjiao (SP 6). Main ear acupoints: Kou (mouth), Fei (lung), Shenmen, and adjuvant ear points: Shenshangxian (adrenal gland), Wei (stomach), Xin (heart), Neifenmi (endocrine), Qiguan (trachea), Gan (liver). The treatment was given once daily, 6 sessions constituting one course, and the therapeutic effects were observed after 4 courses. RESULTS: Thirty-six were cured, 12 cases were effective, 5 cases were ineffective. CONCLUSION: Acupuncture combined with auricular point sticking and pressing has reliable therapeutic effect for smoking cessation, but it is needed that the patient cooperates actively and has the will of smoking cessation, so as to avoid re-taking smoking. PMID- 18405160 TI - [Clinical experiences and thinking of acupuncture for treatment of some difficult and complicated eye's diseases]. AB - The present paper introduces author's some successful experiences on acupuncture treatment of some difficult and complicated eye's diseases in recent years. The author has systematically studied on the law and acupoints of acupuncture for treatment of some difficult and complicated eye's diseases, and worked out therapeutic principles and selected acupoints by means of syndrome differentiation, treating both the principal and the second aspects of a disease, its symptoms and cause at the same time, selecting acupuncture points according to syndromes, combination of Chinese medicine with western medicine, combination of acupuncture with medicine, etc. to strengthen the body resistance and eliminate pathological factors, attaining very satisfactory clinical therapeutic effects. PMID- 18405161 TI - [Meta-analysis on acupuncture for treatment of dementia]. AB - OBJECTIVE: To assess the therapeutic effect of acupuncture on dementia. METHODS: The literatures of acupuncture for treatment of dementia are comprehensively searched in according with the demands of the evidence-based medicine (EBM), which are collected from relevant domestic medical literature databases in the last ten years. Meta-analysis is conducted on the literatures enrolled. RESULTS: Twenty-two randomized controlled trials are included, among them, 19 trials are carried out by Meta-analysis. The total OR is 3.72 [2.73, 5.07], and the funnel plot is approximately symmetry. It is indicated that the curative effect of acupuncture groups is better than the control groups (Z = 8.32, P < 0.00001). CONCLUSION: Acupuncture therapy is effective on dementia according to the domestic clinical literatures. However, the quality of the studies needs further improving and increasing. PMID- 18405162 TI - [Evaluation of literature quality of acupuncture for treatment of herpes zoster and approach to the laws of treatment]. AB - OBJECTIVE: To assess the quality of literature of clinical studies on acupuncture in treatment of herpes zoster. METHODS: The literatures between 1994-2006 were searched by means of electronic retrieval. Type and methodology, general condition, diagnosis of diseases and enrolled and excluded criteria, assessment of sample content, treatment condition, criteria for assessment of therapeutic effects, following-up, etc. in clinical studies are evaluated according to principles and methods of clinical epidemiology and evidence-based medicine. RESULTS: Of the 399 literatures enrolled, only 8 were authentic randomized controlled trials (RCTs), 20 quasi-randomized controlled trials, 66 non randomized concurrent controlled trials and 277 narrative studies, 70 had clear diagnostic criteria, 16 mentioned enrolled or excluded criteria, 287 had clear criteria for therapeutic effects, 107 reported follow-up, 2 had the description of health economical index, 9 reported adverse reaction. CONCLUSION: At present, correct randomization, concealment, blinding and placebo-control, and the RCTs with generally accepted criteria for assessment of diagnosis and therapeutic effects, safety evaluation and rational design of follow-up are needed. It is indicated by preliminary study of the literatures that blood-letting puncture and cupping at Ashi points are main methods for treatment of herpes zoster. PMID- 18405163 TI - [Summarization and analysis on acupuncture-related articles embodied in Medline data-base in 2006]. AB - OBJECTIVE: To understand the situation of studies and development of international acupuncture and moxibustion in 2006 by analysis on the acupuncture related articles embodied in Medline data-base in 2006. METHODS: A total of 881 articles in Medline data-base of the ISI knowledge Web were retrieved by "acupuncture", "moxibustion", "acupressure" and "cupping" as key words, then 761 articles excluding unrelated 120 ones were analyzed and concluded for distribution of the article sources, periodicals source, type, research contents and methods. RESULTS: Most of the articles are from China, Britain and America. The journal of the highest impact factor of 25.8 is Lancet. Acupuncture and moxibustion can treat various diseases, but primarily for relieving pain. CONCLUSION: Acupuncture and moxibustion are still a part of CAM for clinical practice abroad. Most of the experiments confirm the effects of acupuncture and moxibustion, but there are some different views on some diseases. And various countries make a lot work about mechanism research of acupuncture effect, showing the feature of large range. PMID- 18405164 TI - [Preparation and clinical observation of Jiangya Wentie Pian]. AB - OBJECTIVE: To probe into an effective method for treatment of hypertension. METHODS: Eighty cases of hypertension were treated with sticking Jiangya Wentie Plan at Shenque (CV 8) and Yongquan (KI 1) as main. RESULTS: Forty-eight were markedly effective, 29 cases were effective, 3 cases were ineffective, with an total effective rate of 96.25%. CONCLUSION: Acupoint sticking of Jiangya Wentie Plan has obvious therapeutic effect on hypertension. PMID- 18405165 TI - Updated ACCP Guidelines for the diagnosis and management of lung cancer: what are the important changes in recommendations? PMID- 18405166 TI - Weighing the benefits and risks of magnetic resonance scanning of patients with implanted cardiovascular devices. PMID- 18405167 TI - Which should be the first-line drug for newly diagnosed epilepsy? Commentary on the SANAD study. PMID- 18405168 TI - Current European guidelines on cardiovascular disease prevention in clinical practice: an American perspective. PMID- 18405169 TI - Efficacy of pharmacological methods used for treating tobacco dependence: meta analysis. AB - OBJECTIVES: The present study summarizes available evidence describing efficacy of pharmacological methods used in smoking cessation and presents the results of new meta-analyses examining their 12-month efficacy. This work represents part of a larger program examining the efficacy and cost-effectiveness of different methods used in smoking cessation. PATIENTS AND METHODS: The first part of the study included systematic review of literature to identify methods used in smoking cessation with efficacy confirmed on the basis of existing reliable systematic reviews or meta-analyses. In the second stage of the process, for the interventions judged both available in Poland (on the basis of literature search and interviews with healthcare providers) and efficacious, we have performed new meta-analyses designed to establish their 12-month efficacy (continuous or prolonged abstinence). RESULTS: We found that the most comprehensive and up-to date data were available in Cochrane reviews. Meta-analyses of randomised controlled trials performed in the second part of the work showed that adding pharmacological methods of smoking cessation available in Poland, such as nicotine replacement therapy (NRT) and bupropion, to nonpharmacological methods increased the probability of smoking cessation and smoking abstinence for > or =12 months by over 1.5 to about 3 times and the number needed to treat to have one patient stop smoking ranged from 8 to 21. CONCLUSIONS: We confirmed that pharmacological methods of smoking cessation available in Poland, such as NRT and bupropion, added to nonpharmacological methods increase the probability of smoking abstinence and we quantified 12-month effects of these interventions. PMID- 18405170 TI - Diabetes care and self-monitoring of type 2 diabetic patients in a rural district of West-Pomeranian Province. AB - INTRODUCTION: Few surveys conducted in diabetic patients from rural regions show that in these subjects monitoring of diabetes is worse than in patients from urban areas. OBJECTIVES: To assess methods of diabetes care provided for type 2 diabetic patients residing in a rural region and methods of the patient self monitoring of glycemia, blood pressure and foot self-care. PATIENTS AND METHODS: The survey was conducted in a rural district of West-Pomeranian province, in the primary health care center, where 279 type 2 diabetic patients were registered. Out of all patients invited to participate in a questionnaire survey, 168 were enrolled. The mean age of subjects was 67.2 +/- 9.9 years (range 46-91 years), diabetes duration of 8.2 +/- 6.6 years, and body mass index of 32.6 +/- 6.3 kg/m2. Data concerning diabetes care and methods of the patient self-control of glycemia, blood pressure and foot self-care were collected. RESULTS: The majority of patients (62%) were treated only by general practitioners, but 80% reported that they visited their doctors for diabetes treatment once a month. For 90% of subjects the term ,HbA1c" was unknown. Only 40% of patients performed self monitoring of glycemia, 55%--of blood pressure and 34% examined their feet. CONCLUSIONS: The vast majority of patients from a rural region in West-Pomeranian province is treated only by general practitioners. Despite quite frequent medical visits related to diabetes, education of patients is still unsatisfactory, which was demonstrated by patients' lack of knowledge concerning the basic parameter of laboratory monitoring, HbA1c, as well as insufficient self-management of glycemia, blood pressure and infrequent foot exam. PMID- 18405171 TI - Thrombin activatable fibrinolysis inhibitor and other hemostatic parameters in patients with essential arterial hypertension. AB - INTRODUCTION: Hypertension is associated with hemostatic abnormalities and endothelial dysfunction. thrombin activatable fibrinolysis inhibitor (TAFI) is a glycoprotein linking coagulation and fibrinolysis. Objectives. We evaluated TAFI concentrations in patients with essential hypertension in relation to blood pressure. Additionally, we studied TAFI activator, thrombin activity (thrombin antithrombin complexes--TAT, prothrombin fragments F1 + 2), thrombomodulin (TM)- a marker of endothelial cell injury, degree of plasmin generation (plasmin antiplasmin complexes [PAP]), other markers of endothelial cell injury--von Willebrand factor (vWF). PATIENTS AND METHODS: Seventy-two patients with essential hypertension (27 untreated, 13 treated with enalapril (angiotensin converting enzyme inhibitor [ACEI]), 32 with beta-blocker, betaxolol). In every hypertensive patients ambulatory blood pressure measurements and echocardiography were performed. RESULTS: All hypertensive patients did not differ with respect to age, creatinine, fibrinogen, D-dimers. In ACEI-treated patients a significantly higher TAFI concentration was observed when compared to beta-blocker-treated patients. In beta-blocker-treated patients both diastolic and systolic blood pressure were lower than in ACEI treated patients as well as ejection fraction, while serum triglycerides were higher. Diastolic blood pressure correlated significantly with TAFI concentrations in untreated patients (r = 0.27, p < 0.05), and in beta-blocker-treated patients (r = 0.25, p = 0.05), TAFI activity was inversely associated with interventricular septal diameter (r = -0.75, p < 0.01) in patients treated with ACEI. CONCLUSIONS: Elevated TAFI concentrations and enhanced thrombin generation in hypertensive patients may contribute to atherosclerosis progression in this population. Differences in the studied parameters may be due to a small sample size, monotherapy and potential effects of antihypertensive drugs on glicemia, ejection fraction and triglycerides. PMID- 18405172 TI - Optimal treatment for patients after myocardial infarction: some current concepts and controversies. AB - Acute coronary syndromes areone of the most common causes for hospitalizations in developed countries. It is estimated that there are around 1 million patients in Poland after myocardial infarction (MI) requiring medical care. Implementation of optimal treatment in these patients is a significant clinical and social problem. Clinical practice guidelines published by the European Society of Cardiology (ESC) are focused on the crucial tasks such as regular physical activity sufficient to increase exercise capacity, smoking cessation, a Mediterranean diet, intensive control of risk factors for atherosclerosis (hypertension, diabetes, hyperlipidemia), combination pharmacotherapy and in selected cases, coronary revascularization. Of particular importance is that all patients who have had an acute MI should receive optimal treatment with a combination of the drugs that are recommended as class I by the ESC guidelines. Recent registries showed that some of these drugs, including aspirin, clopidogrel, angiotensin converting enzyme inhibitors, beta-blockers and statins, are commonly used in clinical practice in Poland. Recently, it has been highlighted that pharmacological therapy in MI patients should be extended by adding newer agents of the class I recommendations such as omega-3 acid ethyl esters and in selected patients, eplerenone. PMID- 18405173 TI - Is depression a problem in patients with chronic heart failure? AB - Depression is a common psychiatric disorder, characterized by a persistent lowering of mood, loss of interest in routine activities and diminished ability to experience pleasure. There are several depression classification systems and diagnostic tools based on clinical symptoms, i.e. the International Classification of Diseases (ICD-10), the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV), the Hamilton Depression Rating Scale, the Montgomery Asberg Scale and Beck's Depression Inventory. Depression frequently occurs in patients with heart failure, as similar pathophysiological mechanisms of neurohormonal activation, arrhythmia, inflammation and hypercoagulation are present in both these diseases. Prognosis in patients with depression is also affected by insufficient cooperation between a patient and his doctor as regards the lifestyle and medication intake of a patient. Depression is usually accompanied by remission and relapse periods which might be related to the current heart failure status of a patient and despite intensive medical treatment they may recur. Depression is often difficult to diagnose or even left undiagnosed and thus untreated, because its symptoms: fatigue, apathy and decreased exercise tolerance, are common in the general population. Furthermore, safety and efficacy of antidepressant therapy in patients with cardiovascular diseases are not well established. Evidence from clinical trials evaluating the influence of depression behavioral and pharmacological treatment on morbidity and mortality in patients with heart failure is also limited. Taking into account that depression affects prognosis in patients with variety of disorders and common pathological mechanisms present both in depression and heart failure, screening tests for depression should be considered not only in patients with diagnosed heart failure but also those at risk of heart failure development. PMID- 18405174 TI - Vasculopathy and vasculitis in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune connective tissue disease, where vascular lesions are one of the typical symptoms. The pathological process often involves skin vessels, renal glomeruli, the cardiovascular system, brain, lung alveoli, and gastrointestinal tract vessels. This review presents possible adverse mechanisms underlying the cause and effect relationship of various factors causing vascular lesions in SLE patients. The generally accepted hypothesis links vascular damage in SLE with the deposition of immune complexes in the vascular endothelium. The anti-endothelial cell antibodies (AECA), antiphospholipid antibodies and anti-double stranded DNA antibodies present in SLE, that directly or indirectly affect endothelial cells, causing inflammatory damage to the vessel wall, and their role, have been discussed. It has been stressed that although the suggested role of AECA in vasculitis pathogenesis has not been fully established, evidence, however, has demonstrated that AECA is a factor causing endothelial damage in SLE patients. On the other hand, issues concerning cellular adhesion molecules which enable leukocyte adhesion and rolling along the endothelial cell surface, and their extravascular migration, focus on the role they may be playing in SLE patients with vasculitis. A potential role of soluble forms of adhesion molecules, pentraxin 3, medications, infections in the pathogenesis of this disease has also been shown. Special attention has been given to the role of type 3 hepatitis virus in vascular damage in SLE. PMID- 18405175 TI - Still's disease in patient with silicone breast implants: case report. AB - Still's disease is a systemic manifestation of juvenile arthritis that rarely occurs in adults and therefore is often forgotten in differential diagnosis of fever of unknown origin. The case presented here shows diagnostic difficulties in a patient with silicone breast implants and fever with Still disease. The cause effect relationship between breast implants and Still disease is not certain, however, this possibility should be considered when a patient with fever resistant to antibiotics and with silicone implants is admitted to the hospital. PMID- 18405176 TI - Spinal epidural abscess penetrating into retroperitoneal space in patient with diabetes mellitus type 2: early diagnosis and treatment requirement. AB - Spinal epidural abscess (SEA) is a rare condition with very serious prognosis. Predisposing factors for SEA include bacterial infections, immunocompromised states such as diabetes mellitus, intravenous drug abuse, alcoholism, AIDS, as well as spinal surgery and modern techniques of epidural anesthesia. The most common causative agent for SEA is Staphylococcus aureus. The typical clinical signs of SEA are back pain, fever and neurologic dysficit. Magnetic resonance (MR) of the spine and vertebral column is the best imaging diagnostic method in suspected cases. Emergency surgical decompression combined with intravenous antibiotics is the therapeutic method of choice. Conservative treatment may be appropriate in selected patients. Unless the typical presentation of SEA correct diagnosis of this illness is often overlooked and not considered initially. It delays suitable management and leads to poor outcome. We report a classic case of SEA in a woman with a history of diabetes mellitus. PMID- 18405178 TI - Oncological organizations and institutions in Poland before the second World War. AB - During the first four decades of the 20th century many oncological organizations and institutions were established in Poland. On 6 June, 1906 the Committee for Cancer Research and Control was founded. After the year 1918 when Poland regained its independence many more cancer control organizations came into being. The organizations created at that time were: the Polish Committee for Cancer Control (Warsaw 1921), the Cancer Control Society of Lodz (Lodz 1927), the Vilnius Committee for Cancer Control (Vilnius 1931), the Polish Cancer Control Institute Association in Lviv (Lviv 1929), the Polish Cancer Control Association (Warsaw 1938). These cancer control organizations undertook multiphase activities (prophylactic, educational, organizational, scientific and research) by creating outpatient and inpatient oncological care institutions, on which base numerous scientific and research papers were produced. The effect of that work was the creation of the Radium Treatment Institute in Lodz and the Research and Therapeutic Institute for Cancer Control in Vilnius, as well as founding oncological hospital wards and clinics. The most important achievement was building and opening the very modern Radium Institute in Warsaw. The initiator of the Institute, which in 1939 had 90 beds on its disposal, was Maria Sklodowska Curie herself. Cancer control organizations members and activists conducted many propaganda actions on malignant diseases, shared they observations and research findings during meetings, conferences and assemblies in Poland and abroad. PMID- 18405177 TI - Stanislaw Skalski (1870-1937): an eminent Lodz doctor. AB - : Stanislaw Skalski was an eminent Polish specialist in internal medicine. He lived at the turn of the 20th century. He received his medical doctor's diploma at the Kiev University. His professional career and social activity were associated with Lodz where he performed a variety of management functions, among others he held the office of the Chancellor of the City School Board and the Head of the Health Department of the Provincial Office of Lodz. It was the accomplishment of doctor Stanislaw Skalski to introduce obligatory anti-smallpox vaccinations in Lodz (1914) and to promote activities for the improvement of school hygiene and children health. PMID- 18405179 TI - Cutaneous T-cell lymphoma. Preface. PMID- 18405180 TI - Introduction: cutaneous T-cell lymphoma (CTCL)--classification, staging, and treatment options. PMID- 18405181 TI - Long-term bexarotene monotherapy in large cell CD30+ pleomorphic T-cell lymphoma. PMID- 18405182 TI - Denileukin diftitox therapy for patients with tumour-stage mycosis fungoides. PMID- 18405183 TI - Bexarotene monotherapy for patients with advanced stage mycosis fungoides. PMID- 18405184 TI - Treatment of Sezary syndrome with bexarotene after IFNalpha and methotrexate failure. PMID- 18405185 TI - Bexarotene plus PUVA plus radiation combination therapy for mycosis fungoides. PMID- 18405186 TI - Bexarotene salvage therapy in a patient with refractory erythrodermic mycosis fungoides after failure on interferon/PUVA and methotrexate. PMID- 18405187 TI - Bexarotene monotherapy for epidermotropic CD8+ CTCL. PMID- 18405188 TI - Treatment of refractory stage IV mycosis fungoides with bexarotene monotherapy. PMID- 18405189 TI - Bexarotene combination therapy for patients with Sezary syndrome. PMID- 18405190 TI - Bexarotene monotherapy for patients with refractory Sezary syndrome. PMID- 18405192 TI - Climate change and human health. PMID- 18405193 TI - Uganda health care. PMID- 18405191 TI - Bexarotene monotherapy for patients with refractory early stage mycosis fungoides. PMID- 18405194 TI - Vision and strategy for ubiquitous health care: the end of business as we know it. AB - Health services and the clinical enterprise have entered a new era involving an increasing amount of economic, service and research activity across rather than within the boundaries of traditionally defined organizations. These new inter organizational arrangements present unprecedented challenges and opportunities. How they are designed and managed will have profound consequences on the quality, safety and cost of health care. Building on knowledge gained from ongoing activities within health services as well as other sectors, but recognizing the unique challenges in the provision of health care, guidelines are presented that may be of benefit to those involved in emerging partnerships, networks and alliances. PMID- 18405195 TI - Global health diplomacy: training across disciplines. AB - Global health diplomacy is an emerging discipline where health is incorporated into traditional diplomacy as a foreign policy tool. As, with any new area, there is a need for training. This article sets out what global health policy involves and the current academic response to providing training for it. PMID- 18405196 TI - Korean hospital design, state of the art. AB - This paper analyzes the architectural characteristics and trends and suggests some tasks for better hospitals. Hospital design has been improved qualitatively in architectural form and interior. Although overall forms of most buildings have a vertical concept, horizontal concept hospitals have been attempted. In ward plan, various shapes have appeared and interior space has been more dynamic. Special clinics and centres have been established and operated to meet the high level of patients' demand. Suggestions for better hospital could be the increase of private rooms, decreasing the size of the nursing unit, the use of the horizontal concept, supplement of safety-privacy-hygiene, and some sensory factors. PMID- 18405197 TI - Creating competent health-care specialists: the Swiss school of public health. AB - The Swiss health-care system (the second most expensive worldwide) is fragmented into 26 cantonal authorities for a population of 7.5 million. Cantons differ in policy, legislation and structure. Health insurance is compulsory although contributions vary greatly between cantons. A recent report by the Organization for Economic Cooperation and Development (OECD) pointed out that weak governance has led to a system in which "efficiency can be improved", "a broader legal framework for health promotion and disease prevention is overdue", and "equity is not guaranteed". To a certain extent, this situation is due to the scarcity of specialists who know how to judge and respond to health needs and who also understand the complexities of financial flows and the effect of policy interventions in complex systems. As in most countries, health economics and public health have developed independent training programmes, mostly without any coordination or cooperation. Health services therefore are often managed by lawyers or business economists who apply free-market instruments to this regulated system. In the Swiss context, this leads to ever-increasing costs and inequalities without evaluation of potential health gains. PMID- 18405198 TI - The importance of militaries from developing countries in global infectious disease surveillance. AB - Military forces from developing countries have become increasingly important as facilitators of their government's foreign policy, taking part in peacekeeping operations, military exercises and humanitarian relief missions. Deployment of these forces presents both challenges and opportunities for infectious disease surveillance and control. Troop movements may cause or extend epidemics by introducing novel agents to susceptible populations. Conversely, military units with disease surveillance and response capabilities can extend those capabilities to civilian populations not served by civilian public health programmes, such as those in remote or post-disaster settings. In Peru and Thailand, military health organizations in partnership with the military of the United States use their laboratory, epidemiological, communications and logistical resources to support civilian ministry of health efforts. As their role in international affairs expands, surveillance capabilities of militaries from developing countries should be enhanced, perhaps through partnerships with militaries from high-income countries. Military-to-military and military-to-civilian partnerships, with the support of national and international civilian health organizations, could also greatly strengthen global infectious disease surveillance, particularly in remote and post-disaster areas where military forces are present. PMID- 18405199 TI - Sharing and management of EHR data through a national archive: experiences from Finland. AB - The management of Electronic Health Records is a complex business ranging beyond just digital archiving. This article looks at the challenges involved, the different models for EHR data archiving and the impact on health-care provider systems, as well as expected benefits. Particular attention is paid to the Finnish system and its experiences. PMID- 18405200 TI - War-scarred Iraqis face health burdens in foreign lands. PMID- 18405201 TI - Evolution of evidence-based medicine and implications for pharmacy & therapeutics committees. PMID- 18405202 TI - Correlation of HEDIS diabetes health plan score with utilization of diabetes medications. AB - This study sought to determine the correlation between a health plan's Health Plan Employer Data and Information Set (HEDIS) score for glycated hemoglobin (HbA1c) control in patients with diabetes and its utilization of analog insulin, human insulin, and oral drug therapy as determined by the share of prescriptions of each therapy. Prescription volumes were tracked for four categories of diabetes drug therapy: (1) analog insulin, (2) human insulin, (3) single-source brand oral products, and (4) multisource generic oral products, for the three months ending January 2005 and January 2006 and matched to the 2004 and 2005 HEDIS scores. A correlation analysis conducted between the HbA1c-based HEDIS score and the prescription share of each drug category found a favorable and statistically significant (P < .0001) correlation between plan HbA1c HEDIS score and plan prescription share of analog insulin in both 2004 and 2005. The correlation between HEDIS scores and human insulin was not statistically significant. Unfavorable correlations were found between HEDIS scores and both the single-source brand and the multisource generic oral category prescription shares, although these correlations were found to be significant only for the single-source products in 2005. PMID- 18405203 TI - The cost effectiveness of varenicline for smoking cessation. AB - A decision-analysis model was developed to evaluate health and economic effects of varenicline compared with other smoking-cessation aids for private health plans, Medicaid plans, or employee populations. Use of varenicline is projected to increase the number of successful smoking cessations after 10 years by approximately 14% compared with bupropion, 25% compared with nicotine patches, and 38% when compared with no pharmacologic aids. Varenicline use also results in immediate health care cost savings, compared with use of bupropion and savings within two years compared with nicotine patches or no aids. Comparing varenicline with no aids, the cost effectiveness of varenicline at two years ranged from $648 per additional cessation in the private health plan model to $836 per additional cessation in the Medicaid model. Employers often experience additional savings from decreased absenteeism and increased productivity, with combined savings in health care plus workplace costs associated with varenicline use of $165 to $457 per smoker over two years. PMID- 18405204 TI - The direct and indirect cost burden of atopic dermatitis: an employer-payer perspective. AB - The goal of this study was to quantify the incremental direct medical and indirect work-loss costs associated with patients diagnosed with atopic dermatitis (AD). A de-identified administrative claims database was used comprising 5.1 million covered beneficiaries from 31 Fortune 500 self-insured employers between 1998 and 2005. Patients with at least two AD diagnosis claims (N = 13,749) were compared with three matched controls (based on yr of birth and gender) with no AD diagnosis (N = 41,247). In addition, a multivariate two-part regression analysis was used to isolate the cost increase attributable to AD by controlling for confounding factors such as age, gender, health plan type, comorbidities, organ transplantation, industry of employer, region, and year. Direct medical and indirect work-loss costs for the AD group were higher on average by $88 and $64 per patient per month, respectively (both P< .001). After multivariate adjustment, the total incremental cost per patient per month for the AD group was $83 (direct: $52, P< .001; indirect: $31, P< .001). Employer-payers experience a significant annual cost burden of $991 per patient attributable to AD. Employee disability and increased sick days account for 38% of the cost burden. PMID- 18405205 TI - The growing issue of medication-related adverse events. PMID- 18405206 TI - Pharmacy measures to improve medication use through health-literacy principles. AB - More than 90 million Americans have trouble understanding basic health information. Patients with inadequate health literacy encounter problems when trying to understand information related to medication. Consequently, this population has a greater chance of not using pharmaceuticals properly, resulting in poor clinical outcomes. Health care professionals, particularly pharmacists, should improve communication efforts with low-health literacy patients to achieve optimal results. This review examines the relationship between health literacy and medication use with the goals of helping the reader understand how to improve medication adherence and decrease adverse events from improper medication use. PMID- 18405207 TI - Disease-management programs for special populations. PMID- 18405208 TI - An evaluation of Washington's Medicaid disease-management program. AB - In 2002, Washington State Medicaid implemented a disease-management program for clients with diagnoses of asthma, chronic obstructive pulmonary disease, heart failure, and diabetes. The program represented a unique attempt to manage disabled clients in a fee-for-services environment, and at its onset, was one of the first statewide programs in the United States. This article reviews the effectiveness of the program based on the results from two independent evaluations. Results of cost-savings measurements and health outcomes are presented for each of the conditions. These results were used to make program changes, which began in 2007. PMID- 18405209 TI - Is the oral health strategy a cause for hope? PMID- 18405210 TI - Creating smiles. PMID- 18405211 TI - The dental patient with a congenital bleeding disorder. AB - Congenital bleeding disorders account for approximately one in 10,000 births. Dentists are often anxious about delivering treatment to this special group of patients. In the Irish Republic, patients with inherited bleeding disorders have their dental care co-ordinated centrally at the National Centre for Hereditary Coagulation Disorders (NCHCD), St James's Hospital, Dublin. Dental care is normally integrated with routine outpatient haematological appointments. This ensures regular monitoring of oral health and the early treatment of any hard/soft tissue pathology. This article describes, in simple diagrammatic form, the normal coagulation mechanism (Figures 1 and 2), explains common coagulation terms (Appendix 1), and examines the three most common congenital bleeding disorders: haemophilia A, haemophilia B, and von Willebrand disease. General recommendations based on the current literature are provided with respect to procedures that are appropriate to perform in a general dental practice setting. Although not discussed in this article, it is important to note that non coagulation bleeding disorders also exist. These include: hereditary haemorrhagic telangiectasia; blood vessel wall defects resulting from connective tissue disorders such as Marfan syndrome and Ehlers-Danlos syndrome; and, platelet disorders such as Bernard-Soulier syndrome, resulting in defective platelet adhesion. PMID- 18405212 TI - Oral health and orthodontic considerations in children with juvenile idiopathic arthritis: review of the literature and report of a case. AB - Juvenile idiopathic arthritis (JIA) is a severe disease of childhood, which comprises a diverse group of distinct clinical entities of unclear aetiology. Some abnormality of the immune system is present in all JIA cases. In its most severe clinical form, JIA may show localised and/or systemic complications, including functional impairment of the affected sites. This may result in variable growth and developmental anomalies. In many JIA cases, where the temporomandibular joint (TMJ) is affected, mandibular growth may be restricted, thus leading to the development of mandibular hypoplasia and/or retrognathism. As a result, it is not uncommon for JIA patients to present with skeletal Class II and open bite malocclusions. Furthermore, in JIA cases with unilateral TMJ involvement, craniofacial asymmetry may occur. In such cases, early orthodontic intervention facilitates both the skeletal and the occlusal rehabilitation. Increased prevalence of dental caries and periodontal disease in JIA cases may be attributed to a combination of aetiological factors, including difficulties in executing good oral hygiene, unfavourable dietary practices and side effects from the long-term administration of medication. In addition, an association between periodontal disease and JIA has been reported based on their similar pattern of clinical disregulation of the inflammatory process. This paper presents a brief description of JIA, with special reference to dental health and orthodontic treatment considerations. In addition, a case is presented where the appropriate orthodontic intervention led to the establishment of a normally functioning, as well as an aesthetically pleasing, occlusion. PMID- 18405213 TI - Radiation safety of the patient. PMID- 18405215 TI - Taxes, taxes and less taxes? PMID- 18405214 TI - Re. About Second District. PMID- 18405217 TI - Why a dental practice needs a slice of corporate America. PMID- 18405216 TI - Methamphetamine abuse and "meth mouth". AB - Dental management for the patient who abuses drugs is always a challenge. The numbers of patients abusing methamphetamines appears to be increasing. The dentist needs to be aware of the clinical presentation and medical risks presented by these patients and to attempt to get the patient to seek professional help. Additionally, special attention will be necessary for the high prevalence and severity or oral manifestations including: rampant caries, enamel erosion, xerostomia, bruxism and muscle trismus. PMID- 18405218 TI - Error is human. PMID- 18405219 TI - HIV testing: an update. PMID- 18405220 TI - Sex, human papilloma virus infection, and head and neck cancer. PMID- 18405221 TI - Adulterants and drugs-of- abuse testing: an update. PMID- 18405222 TI - Wireless, paperless, anything but careless. PMID- 18405223 TI - Basics of home glucose monitoring. PMID- 18405224 TI - Using voicemail effectively. PMID- 18405225 TI - Sue over review? PMID- 18405226 TI - Laying the foundation for HIEs. PMID- 18405227 TI - [Insurance medicine risk assessment--also required in the future]. PMID- 18405228 TI - [Contentious diseases--a medico-social phenomenon from an insurance medicine perspective]. AB - A group of illnesses that are difficult to assess objectively, comprising such conditions as fibromyalgia, chronic fatigue syndrome, attention-deficit hyperactivity disorder, whiplash injury, and last but not least a multitude of somatoform disorders, has become a growing concern to Western health care systems and insurance industries. Thus far, the medical literature has failed to provide informative overviews of this group, which at first glance admittedly seems to be rather heterogeneous. If at all, the disorders have been grouped together under the term ,,controversial illnesses" to differentiate them from other diseases. The insurance industry - and claims departments, in particular - are increasingly having to deal with this rapidly growing phenomenon, which affects not only life business, but also health, worker's compensation and motor third-party liability. When paying compensation and settling claims, insurers are often left with a feeling that the illness may have been ,,imaginary" or aggravated. Is there a common basis for this new disorder mega-trend - independent of the recognition of the conditions by medical associations? This article aims at providing an overview of the common characteristics of the group of disorders, including a description of the key physical, psychological and social aspects. In particular, it is intended to deepen insurers' understanding of the risks arising from social change. The article also examines the disorder prevalence in Western societies and the possible causes of the significant increase. PMID- 18405229 TI - [Malingering of post-traumatic stress disorder]. AB - Post-traumatic stress disorder is a frequently diagnosed disorder. Expert assessments of claimants with PTSD symptoms have become a common challenge for physicians in civil lawsuits. Since diagnosis of PTSD relies primarily on the patient's subjective report, this syndrome is particularly vulnerable to malingering. For this reason, it is important for physicians to try to identify individuals with true PTSD and differentiate them from those who malinger the disorder. This paper outlines the general problems and assessment strategies involved in the detection of malingering. A special focus is placed on the typical presentation of malingered PTSD symptoms. PMID- 18405230 TI - [Coping with crisis as an important factor in the rehabilitation process]. AB - Accidents and serious illnesses are inevitably associated with an emotional crisis, based on the fact that they are followed by significant consequences. Coping with this crisis requires enormous mental adaptation. This active coping process is typically characterised by three phases: the shock phase, the coping phase in the narrow sense and the phase of successful coping versus chronification. In order to ensure a positive rehabilitation process, successful coping with the illness is necessary. It is therefore absolutely crucial to analyse individual coping styles and possible obstacles thoroughly from a psychological point of view. In order to prevent a process of chronification, relevant psychosocial factors should be considered along with the medical and occupational aspects early in the rehabilitation process. Ultimately, psychological stability is the prerequisite for successful rehabilitation. PMID- 18405231 TI - [Patients requiring high healthcare spending]. AB - Data from private insurance companies make it possible to analyse how healthcare spending is distributed across individuals, how it depends on the age of the people and how it changes over time. Within age groups, healthcare spending is less concentrated if recipients are older. Over the analysed period of time, a considerable levelling of expenses takes place. These findings lead to the conclusion that the ageing population will result in a greater and more evenly spread utilisation of healthcare facilities. PMID- 18405232 TI - [Diagnosis and treatment methods of alternative medicine in private health insurance--a quantitative analysis]. AB - Diagnosis and treatment methods of alternative medicine are very popular in Germany and are used very often--not only in case of incurable diseases, but for all possible indications. The present study looks into which treatment methods of alternative medicine are used the most. Nutritional supplements and vitamins are way ahead as an alternative medicine, followed by acupuncture, homoeopathy, organotherapy, alternative medicinal phytotherapy, symbiosis controlling, enzyme therapy and TCM (Traditional Chinese Medicine). Treatment methods of alternative medicine are used very often, although there are no studies of a scientifically acceptable level which prove the effectiveness of the therapy. However, the health dangers and side effects of alternative medicine treatments have been scientifically proven. Beyond that, the use of these methods has not been sufficiently examined. PMID- 18405234 TI - [ New quality reports of hospitals]. PMID- 18405233 TI - [Does the obesity "epidemic" stop the life expectancy trend?]. PMID- 18405235 TI - [Psychotherapy in private health insurance]. PMID- 18405236 TI - [Private insurance coverage of laser acupuncture]. PMID- 18405237 TI - Biochemistry and physiology of mammalian secreted phospholipases A2. AB - Phospholipases A(2) (PLA2s) are esterases that hydrolyze the sn-2 ester of glycerophospholipids and constitute one of the largest families of lipid hydrolyzing enzymes. The mammalian genome contains 10 enzymatically active secreted PLA2s (sPLA2s) and two sPLA2-related proteins devoid of lipolytic enzymatic activity. In addition to the well-established functions of one of these enzymes in digestion of dietary phospholipids and another in host defense against bacterial infections, accumulating evidence shows that some of these sPLA2s are involved in arachidonic acid release from cellular phospholipids for the biosynthesis of eicosanoids, especially during inflammation. More speculative results suggest the involvement of one or more sPLA2s in promoting atherosclerosis and cancer. In addition, the mammalian genome encodes several types of sPLA2-binding proteins, and mounting evidence shows that sPLA2s may have functions related to binding to cellular target proteins in a manner independent of their lipolytic enzymatic activity. PMID- 18405238 TI - Effect of donor cell type on complement-dependent cytotoxicity crossmatch outcome for patients immunized against HLA class-II antigens. AB - OBJECTIVES: In the Eurotransplant zone, the crossmatch serum exchange program was established to reduce unnecessary organ shipment, using the complement-dependent cytotoxicity crossmatch as the reference to make the decision. Crossmatching at the donor center dictates whether the transplant should be shipped to the recipient center where a decisive crossmatch would then be done. However, in recent years, the target cell used for the crossmatching has changed from spleen cells to peripheral blood lymphocytes. In this study, we assess the impact of this change on the outcome of complement-dependent cytotoxicity crossmatches for patients immunized against HLA-class II. MATERIALS AND METHODS: The influence of the donor cell type was analyzed by crossmatching unseparated peripheral blood lymphocytes, separated T and B lymphocytes, as well as spleen cells from 12 organ donors with sera from 40 immunized kidney retransplant candidates. Negative sera and sera harboring only anti-HLA class-II antibodies were used as additional controls. We did more than 1200 complement-dependent cytotoxicity crossmatches. RESULTS: Crossmatches with sera containing anti-HLA class-I plus class-II alloantibodies (n=113 per cell type) were positive in 42% of peripheral blood lymphocytes, 72% of spleen cells, and 81% of B cells. Crossmatches with sera containing exclusively anti- HLA class-II antibodies (n=89 per cell type) were positive in 1% of peripheral blood lymphocytes, 30% of spleen cells, and in 31% of B cells. Overall, spleen or separated B cells identified approximately 30% more positive donor-recipient pairs. CONCLUSIONS: The data show that the change from spleen cells to peripheral blood lymphocytes as donor target cells for complement-dependent cytotoxicity crossmatching increased risk of false negative results for patients harboring anti-HLA class-II antibodies. PMID- 18405239 TI - Pediatric liver transplant: results of a single center. AB - OBJECTIVES: Liver transplant in the pediatric population has become an accepted treatment modality for children with end-stage liver disease. In this study, we analyze our experiences with pediatric liver transplant at our center. MATERIALS AND METHODS: Since September 2001, 8 deceased-donor and 96 living-donor liver transplants have been done in 101 children (mean age, 6.7 +/- 5.5 years; range, 2 months to 17 years). The children's charts were reviewed retrospectively. RESULTS: Indications for liver transplant were cholestatic liver disease (n=17), biliary atresia (n=24), Wilson's disease (n=16), fulminant liver failure (n=18), hepatic tumor (n=13), and other (n=13). The median pediatric end-stage liver disease score was 23.1 +/- 11.1 (range, -8 to 48). The median follow-up was 24.2 +/- 19.4 months (range, 1-77 months). Three children underwent retransplant. The main complications were infections (25.9%) and surgical complications (39.5%) (including biliary complications and vascular problems). The incidence of acute cellular rejection was 42.3%. Sixteen children died during follow-up, and, at the time of this writing, the remaining 85 children (85%) were alive with good graft functioning, showing patient survival rates of 90%, 85%, and 83% at 6, 12, and 36 months, respectively. CONCLUSIONS: In conclusion, the overall outcomes of pediatric liver transplantation at our center are quite promising. PMID- 18405240 TI - Liver transplant for hepatocellular carcinoma: experience in a Saudi population. AB - OBJECTIVES: We present our experience with deceased-donor liver transplant and living-donor liver transplant for hepatocellular carcinoma. Between 2001 and 2007, we transplanted 133 organs (84 deceased-donor liver transplants, 49 living donor liver transplants) in 126 patients (4 retransplants). Twenty-three patients had hepatocellular carcinoma (14 deceased-donor liver transplants and 9 living donor liver transplants). MATERIALS AND METHODS: The medical records of these patients were reviewed for recipient clinical, biochemical, and imaging characteristics. Slides of explants were assessed. Overall survival and tumor recurrence states were determined. All characteristics were tested for their prognostic significance. RESULTS: The median age of the patients was 55 years and the median Mayo End-stage Liver Disease score was 16. The alpha-fetoprotein was >or= 400 ng/mL in 4 patients. Histopathology revealed incidental cholangiocarcinoma in 2 patients and a hepatoblastoma in 1. The mean tumor size was 4 cm; the mean number of lesions was 2. Most tumors were graded as well or moderately differentiated; 4 were poorly differentiated. Gross macrovascular invasion was seen in 2 patients, while microvascular invasion was seen in 9. After a mean follow-up of 736 days, overall patient and graft survival rates were 80.9% and 76.2%; overall disease-free patient and graft survival rates were 76.2% and 71.4%. Two patients died of primary graft nonfunction within 1 week of the transplant. Three had tumor recurrence at 10, 13, and 18 months after transplant; 2 of these occurred in patients with cholangiocarcinoma. Two of these 3 died from an advanced tumor within few months. Significant risk factors for recurrence were gross major vessel invasion, microvascular invasion, tumor size, poor histologic differentiation, and absence of pretransplant tumor control therapy. The latter 2, in addition to Mayo End-stage Liver Disease score and preoperative alpha fetoprotein, were independent predictors of mortality. CONCLUSIONS: In our small experience, deceased-donor liver transplant and living-donor liver transplant for hepatocellular carcinoma showed good long-term outcomes. Liver transplant for hepatocellular carcinoma accompanying cholangiocarcinoma had a poor outcome with late tumor recurrence. Use of marginal donors in patients with hepatocellular carcinoma might compromise the outcome in these patients. PMID- 18405241 TI - Prevalence and risk factors of renal dysfunction after liver transplant: a single center experience. AB - OBJECTIVES: Renal dysfunction is one of the most significant complications after liver transplant. It is attributed mainly to nephrotoxicity caused by calcineurin inhibitors. We evaluated the renal functioning in liver transplant recipients alive for at least 6 months after liver transplant. MATERIALS AND METHODS: One hundred seventy patients (108 male [63.5%], 62 female [36.5%]; mean age, 31.4 +/- 13.3 years; age range, 13-61 years) were included in this study. Patients who had undergone a liver transplant between 1994 and 2006 at the Organ Transplantation Center of the Shiraz University of Medical Sciences in Shiraz, Iran, and had been alive for at least 6 months after surgery were included. Data were collected regarding age, sex, body mass index, underlying liver disease, graft type, immunosuppressive medications, serum creatinine levels, and glomerular filtration rate before, 1, and 6 months after liver transplant. Renal dysfunction was defined as a serum creatinine level above 132.6 micromol/L or a glomerular filtration rate less than 60 mL/min/1.73 m2, based on our reference range. Glomerular filtration rate was calculated using the Schwartz formula (glomerular filtration rate mL/min/1.73 m2 = K x Ht (cm) / Cr mg/dL). Data were analyzed with SPSS software. RESULTS: The mean follow-up was 25.9 +/- 23.5 months (range, 6-156 months). The main indications for liver transplant were cryptogenic cirrhosis (n=42), hepatitis B infection (n=34), autoimmune cirrhosis (n=30), Wilson's disease (n=21), and primary sclerosing cholangitis (n=18). The mean pretransplant glomerular filtration rate was 93.7 +/- 35.6 mL/min/1.73 m2. The mean glomerular filtration rates in the first and sixth months after liver transplant were 81.6 +/- 29.3 mL/min/1.73 m2 and 83.6 +/- 32.9 mL/min/1.73 m2. Sex, body mass index, type of immunosuppressive medication, and underlying liver disease were not predictors of renal dysfunction (P > .05). Posttransplant renal dysfunction was significantly more common in older patients (ie, those aged 38.8 years and older) (P = .0001) and those with a family history of renal disease (P < .05). CONCLUSIONS: Renal dysfunction may be a significant problem for patients after liver transplant, and early detection of renal dysfunction in patients after liver transplant is important. Of all the risk factors studied here, only older age and family history of renal disease were correlated with development of renal dysfunction after liver transplant. PMID- 18405242 TI - Nomogram that predicts graft survival probability following living-donor kidney transplant. AB - OBJECTIVES: The goal of this project was to develop a nomogram that predicts the probability of graft survival at 5 years. MATERIALS AND METHODS: From our dataset, 1,581 patients were used to construct a nomogram (modeling group), the remaining 319 patients (testing group) were used for its validation. Initially, the modeling group variables were correlated with graft survival by univariate analysis. Significant factors were subjected to a multivariate analysis using a Cox regression model. The results formed the basis of our nomogram construction. Internal validation was done first by discrimination using the concordance index. Second, the calibration was assessed graphically. And finally, for external validation, the nomogram was used to predict graft survival using the testing group. The predicted probability(s) was compared with the actual survival estimates. RESULTS: Validation of the nomogram yielded a concordance index of 0.77, and the observed correspondence between predicted and actual outcomes suggested a high level of calibration. Nomogram predictions of the testing group revealed no differences in the means of predicted and observed graft survival at 5 years, with a high correlation coefficient and accepted predictive accuracy (concordance index, 0.72). CONCLUSIONS: We developed a well-validated and reasonably precise nomogram for predicting 5-year graft survival. PMID- 18405243 TI - Risk factors for delayed graft function defined as need for dialysis or failure of creatinine to fall by 10% in the first 24 hours after transplant. AB - OBJECTIVES: Delayed graft function after deceased-donor transplant remains a significant clinical problem. The conventional definition of delayed graft function is the requirement of dialysis within the first week after transplant, but this criterion has many problems that have led to many controversies including those of incidence and significance. Therefore, we sought to identify the possible risk factors of delayed graft function and to investigate their effect on short-term graft survival, according to a composite criterion. MATERIALS AND METHODS: We reviewed the records of 94 renal transplants obtained from heart-beating deceased donors done at our center during a 2-year period. Variables related to the donor, recipient, and graft were retrospectively collected. Follow-up was 12 months. Delayed graft function was defined as the need for dialysis or the failure of the creatinine level to fall by 10% during the first 24 hours after transplant. To confirm suspected rejection, protocol biopsies were done, irrespective of graft function, on the seventh and 28th days after transplant, or when indicated to confirm suspected rejection. RESULTS: The overall incidence of delayed graft function was 31.9%. Multivariate analysis showed donor age as a significant independent predictor of delayed graft function (OR=1.05, P = .03, 95% CI: 1.01-1.09), whereas donor hypotension was the only independent risk factor associated with a worse 1-year graft survival rate (OR=4.6, P = .021, 95% CI: 1.3-16.5). No association could be established between delayed graft function, acute rejection, and graft survival. CONCLUSIONS: Advanced donor age is a predictor of delayed graft function defined as the need for dialysis or the failure of creatinine to fall by 10% during the first 24 hours after transplant. Preventing hemodynamic instability should be an important aspect of donor care. PMID- 18405244 TI - Clinicoepidemiologic study of posttransplant diabetes after living-donor renal transplant. AB - OBJECTIVES: We sought to evaluate posttransplant diabetes mellitus with regard to its incidence, risk factors for occurrence, complications, impact on graft function, and impact on patient and graft survival rates. MATERIALS AND METHODS: A total of 1,580 patients received living-donor renal allografts at Mansoura University, Egypt, between March 1976 and November 2004. Of these, 286 recipients developed diabetes after transplant (diabetic group). These patients were matched with 316 kidney transplant recipients who did not develop diabetes after transplant (control group). A complete clinical history was obtained and a clinical examination was done. Laboratory analyses including urine analysis, complete blood count, total serum cholesterol, fasting and 2-hour postprandial plasma glucose, Hb A1c, serum creatinine, and creatinine clearance were obtained in all patients. In each patient, presence of hepatitis B and C was determined with polymerase chain reaction, and a graft biopsy was obtained to diagnose renal allograft rejection. RESULTS: The onset of diabetes mellitus among our recipients occurred primarily during the first 6 months after transplant (in 52.4% of the patients). Significant correlations were found between posttransplant diabetes mellitus and the recipients' age (P = .0001), obesity (P = .001), positive family history of diabetes mellitus (P = .001), hepatitis C virus infection (P = .039), cumulative dose of steroids in the first 3 months (P = .047), and calcineurin inhibitor-based immunosuppressive therapy (P = .001). Moreover, posttransplant diabetes mellitus significantly affected rates of coronary heart disease (P = .001), hypertension (P = .02), and hypercholesterolemia (P = .001). Graft survival was similar in both groups until 15-year follow-up, at which time graft survival began to decrease in patients with diabetes mellitus compared with those without diabetes mellitus (43.5% vs 53.6%, P = .013). Similarly, patient survival was similar until 8-year follow-up, at which time survival rates began to decline in patients with diabetes as compared with patients without diabetes (79.9% vs 86.1%, P = .001); this trend continued to the 15-year follow-up (60.6% vs 77.8%, P = .001). CONCLUSIONS: Posttransplant diabetes mellitus is a major problem that endangers patient and graft survival. In our population, the incidence of posttransplant diabetes mellitus was 18.2%. Further studies are recommended to screen for patients with impaired fasting glucose and impaired glucose tolerance for prediction, early detection, and better management of posttransplant diabetes mellitus. PMID- 18405245 TI - Rescue immunosuppressive therapies in living-related renal allotransplant: a long term prospective randomized evaluation. AB - OBJECTIVES: The majority of our patients are maintained on prednisolone, cyclosporine, and azathioprine as primary immunosuppression. In the presence of repeated episodes of acute rejection, this maintenance immunosuppressive regimen is increased by replacing cyclosporine with tacrolimus or azathioprine with mycophenolate mofetil. To the best of our knowledge, there are no available data among living-related renal allotransplants that evaluate the long-term efficacy and safety of these rescue immunosuppressive therapies. Therefore, we sought to evaluate the long-term efficacy and safety of rescue immunosuppressive therapies among living-related renal allotransplant recipients. MATERIALS AND METHODS: We reviewed the long-term follow-up data of 212 renal transplant recipients at the Urology and Nephrology Center Mansoura University in Mansoura, Egypt, who had been maintained on a primary immunosuppressive protocol that included prednisolone, cyclosporine, and azathioprine. Patients were randomized at a ratio of 1:2 to receive more-intensive maintenance immunosuppression by replacing cyclosporine with tacrolimus in 65 patients (group TAC) and replacing azathioprine with mycophenolate mofetil in 147 patients (group MMF). RESULTS: We found no statistically significant difference between the 2 groups regarding rejection-free patients or those who experienced 1 or more episodes of acute rejection (P > .5). In group TAC and group MMF, graft survival rates were 87.3% and 96.3% at 2 years and 78.7% and 80% at 5 years, respectively (P = .07). The corresponding patient survival rates were 98.4% and 98.5% at 1 year, 98.4% and 97.7% at 2 years, and 94.4% and 94.4% at 5 years, respectively (P = .65%). There were more patients with diabetes and serious bacterial infections in group TAC than there were in group MMF (P = .001 and .04, respectively). CONCLUSIONS: Conversion from cyclosporine to tacrolimus or from azathioprine to mycophenolate mofetil is a safe, equipotent rescue especially with repeated acute rejections. However, mycophenolate mofetil rescue therapy was more beneficial regarding graft survival. PMID- 18405246 TI - Polymorphism of the methylenetetrahydrofolate reductase C677T gene with chronic allograft nephropathy in renal transplant recipients. AB - OBJECTIVES: This study sought to investigate the frequency of the 5, 10 methylenetetrahydrofolate reductase gene (MTHFR C677T) in 127 patients (77 with chronic allograft nephropathy and 50 with normal renal function) who had undergone a renal transplant at least 20 months earlier to define the risk factors for chronic allograft dysfunction. Fifty healthy subjects served as controls. MATERIALS AND METHODS: Genotypes were determined using polymerase chain reaction followed by restriction fragment length polymorphism analysis. The restriction enzyme for the MTHFR C677T variants was HinfI. RESULTS: No statistically significant differences were seen between the allelic and genotypic distribution of the MTHFR polymorphism. CONCLUSIONS: Additional studies with larger sample sizes are needed to define the influence of MTHFR C677T genotyping on clinical outcomes in renal allograft recipients. PMID- 18405247 TI - Impact of sirolimus treatment in kidney allograft recipients with prolonged cold ischemia times: 5-year outcomes. AB - OBJECTIVES: Sirolimus, an effective and non-nephrotoxic immunosuppressant, may have an antiproliferative effect on renal tubular cells and increase their apoptosis, thus hindering the recovery of an injured kidney. The aim of this study was to evaluate the impact of combined sirolimus and cyclosporine therapy on the incidence and duration of delayed graft function and long-term graft function. MATERIALS AND METHODS: The study group consisted of 23 renal transplant recipients treated with a sirolimus-cyclosporine-prednisone regimen (sirolimus group). The reference group was composed of 23 patients treated with azathioprine cyclosporine-prednisone. Because of a long cold ischemia time, all the patients were at high risk of developing delayed graft function. RESULTS: There was an equal frequency of delayed graft function in the sirolimus group compared with the reference group (39% vs 34.8%). The duration of delayed graft function was longer in sirolimus group compared with the reference group (21.2 +/- 12.2 days vs 6.8 +/- 2.5 days) (P < .004). The serum creatinine level at the 12th month was higher in patients with delayed graft function than it was in the remaining patients, independent of the immunosuppression protocol. One and 5-year graft survival rates were 100% and 87% in the sirolimus group, and 95% and 74% in the reference group. The 5-year patient survival rate was 100% in both groups. CONCLUSIONS: Sirolimus significantly retards the recovery from posttransplant renal failure, but it does not increase the incidence of delayed graft function. Sirolimus therapy should be initiated after recovery from posttransplant renal failure. Sirolimus treatment is beneficial for long-term graft survival. PMID- 18405248 TI - Time of onset, viral load, relapse, and duration of active cytomegalovirus infection in bone marrow transplant outcomes. AB - OBJECTIVES: Active cytomegalovirus infection remains a major problem for bone marrow transplant recipients. If not quickly diagnosed and treated, it can evolve into cytomegalovirus disease, which represents a life-threatening complication. In this work, we sought to evaluate the interactions between clinical complications after bone marrow transplant and factors associated with active cytomegalovirus infection. MATERIALS AND METHODS: We evaluated 91 allogeneic bone marrow transplant recipients (35 female, 56 male; median age, 20 years; age range, 3-47 years) for malignant and nonmalignant hematologic diseases. Active cytomegalovirus infection was monitored using pp65 cytomegalovirus antigenemia and a semiquantitative cytomegalovirus polymerase chain reaction. Cytomegalovirus end-organ disease was defined as an association between compatible signs and symptoms (dyspnea, hypoxia, and diarrhea) and detection of cytomegalovirus (>or= 2,000 cytomegalovirus genome copies/mL) by hybrid capture assay in tissue biopsy. Variables were compared using the chi-square and Fisher exact tests. Time of death after bone marrow transplant was plotted using the Kaplan-Meier method. A Cox regression model was used for multivariate survival analysis with 95% confidence limits. RESULTS: Sixty-four patients experienced active cytomegalovirus infection, 26 had acute graft-versus-host disease, and 11 had cytomegalovirus diseases. The overall survival rate at 4 years was 83.52%. On multivariate analyses, cytomegalovirus disease (hazard ratio = 15.9, P = .001) and age older than 18 years (hazard ratio = 8, P = .18) were the only independent negative prognostic factors for overall survival. Occurrence of acute graft versus-host disease was increased by early active cytomegalovirus infection (P = .03) and represents a significant factor for active cytomegalovirus infection recurrence (P = .01). Viral load as quantified by antigenemia and cytomegalovirus DNA in the patients' peripheral blood leukocytes was significantly associated with clinical complications. CONCLUSIONS: Active cytomegalovirus infection interacts significantly in several ways with graft-versus-host disease and others infections. Acute graft-versus-host disease increases the chances of a poor outcome, especially of acquiring cytomegalovirus disease. Cytomegalovirus disease constitutes a significant independent risk factor for death after bone marrow transplant. PMID- 18405249 TI - Interleukin-10 gene polymorphism in bone marrow transplant recipients. AB - OBJECTIVES: Graft-versus-host disease is the main complication after hematopoietic stem cell transplant, occurring even after donor and recipient human leukocyte antigen matching, apparently because of donor/recipient minor histocompatibility antigen mismatches and cytokine polymorphisms. Interleukin-10 suppresses several activities of the immune response by inhibiting T helper 1 and T helper 2 cells. These properties suggest that interleukin-10 could act as a suppressive mediator and prevent graft-versus-host disease. This study evaluates the association between the interleukin-10 promoter gene polymorphism and transplant outcomes among 18 recipients of cytokine-mobilized peripheral blood stem cells from human leukocyte antigen-matched sibling donors. MATERIALS AND METHODS: We analyzed 3 single nucleotide polymorphisms in the proximal region of the interleukin-10 promoter gene (-1082/-819/-592) by the amplification refractory mutation system and polymerase chain reaction-restriction fragment length polymorphism methods. Eighteen donors and their recipients who had undergone an allogeneic peripheral blood stem cell transplant at the Bone Marrow Transplant Center in Nemazi Hospital (Shiraz, Southern Iran) between September 2005 and September 2006 were enrolled. RESULTS: The GCC haplotype (1082G/819C/592C) was predominant in both the donor and the recipient, but no significant correlations were present between the GCC haplotype in either the donor or the recipient and the risk of acute graft-versus-host disease (P = .56). CONCLUSIONS: The interleukin-10 promoter gene polymorphism was found not to be associated with acute graft-versus-host disease in patients after an allogeneic peripheral blood stem cell transplant from human leukocyte antigen-matched sibling donors. Additional studies with larger samples are necessary to further define the influence of interleukin-10 on the immune response after bone marrow transplant. PMID- 18405250 TI - Sinus bradycardia associated with daclizumab in liver transplant recipients: report of 3 cases. AB - Daclizumab is a commonly used immunosuppressive agent for prophylaxis of solid organ rejection. Although rare, the cardiovascular adverse effects of daclizumab include sinus tachycardia, hypotension, and hypertension. Here, we report 3 patients who developed significant and prolonged sinus bradycardia after receiving daclizumab following orthotopic liver transplant. Daclizumab should be considered a possible cause of bradycardia following its administration in orthotopic liver transplant. PMID- 18405251 TI - Justifying a third pancreas transplant: a case report. AB - Early pancreas graft failure after simultaneous pancreas-kidney transplant can occur in up to 20% of recipients. Results after pancreas retransplant continue to improve, with results comparable to primary pancreas transplants. We describe an unusual case of a third pancreas transplant in which a remnant of a previous arterial Y-graft was used for the arterial anastomosis, and we discuss the factors used to justify the decision to do a third pancreas transplant. PMID- 18405252 TI - Uterus transplant techniques in primates: 10 years' experience. AB - Preserving and restoring fertility are important quality-of-life outcomes for cancer survivors and others who cannot bear children. Transplantation has advanced sufficiently to allow nonvital organ transplant to become a reality. Together, research on reproductive organs and transplantation has resulted in live human births from ovary transplants used to treat absolute ovary factor infertility. Each year, approximately 5,000 women under the age of 25 years undergo a hysterectomy. From various other causes, approximately 9 million women of childbearing age in the United States currently have absolute uterine factor infertility due to the absence of the uterus. Research on uterus transplantation has been reported for decades from multiple institutions around the world. Our group has been involved in the area for more than 10 years. To encourage other investigations in this area, we summarize our results in primates as a proof of concept that uterus transplants can be theoretically done. PMID- 18405253 TI - Evidence-based practice: the debate. PMID- 18405254 TI - Evidence-based practice: implications and concerns. AB - AIMS: The aim of this paper was to undertake a brief critical appraisal of evidence-based practice (EBP) as it is currently perceived in health care settings. BACKGROUND: The past two decades have seen EBP become increasingly important in health care planning, clinical thinking, and choice of treatments. It is based on scientific rationalism and adherents claim that decisions based on EBP are superior to those based on other approaches to care. Concerns are now being expressed that positivistic approaches to health care fail to take into account people's preferences, their internal resources and their personal understandings of health and wellbeing. It has been argued that there may be multiple types of evidence, all of which have a part to play in the formulation and execution of health care. METHODS: After a literature search, this paper argues that whereas EBP may be useful in treating conditions that have a biological cause, it may be less helpful in understanding and treating conditions that have their origins in the social, psychological or spiritual domains. RESULTS: The nature, strengths and limitations of evidence-based practice is discussed in this paper. Nurses are encouraged to develop the critical skills of evaluating EBP in the lives and experiences of the people they care for. CONCLUSIONS: Evidence-based practice has a part to play in improving the treatment provided for patients. Nonetheless, nurses should be aware of other kinds of evidence, and appreciate that any single approach to determining care, no matter how popular, is likely to lead to a service that does not truly meet the complex individual needs of patients. Implications for nursing management In order for evidence-based practice to be safe, the nursing workforce must be able to evaluate the strength and relevance of research findings, and be able to understand that there are different kinds of evidence which should be called upon in order to respond sensitively and appropriately to the preferences of patients. A responsive workforce embraces multiple ways of thinking, respects different paradigms of care, and is able to respond to and respect the forms of care people value and seek. PMID- 18405255 TI - Nursing Best Practice Guidelines: reflecting on the obscene rise of the void. AB - AIM(S): Drawing on the work of Jean Baudrillard and Michel Foucault, the purpose of this article is to critique the evidence-based movement [and its derivatives - Nursing Best Practice Guidelines (NBPGs)] in vogue in all spheres of nursing. BACKGROUND: NBPGs and their correlate institutions, such as the Registered Nurses' Association of Ontario (RNAO) and 'spotlight' hospitals, impede critical thinking on the part of nurses, and ultimately evacuate the social, political and ethical responsibilities that ought to distinguish the nursing profession. EVALUATION: We contend that the entire NBPG movement is based on the illusion of scientific truth and a promise of ethical care that cannot be delivered in reality. We took as a case study the Registered Nurses' Association of Ontario (RNAO), in the province of Ontario, Canada. KEY ISSUES: NBPGs, along with the evidence-based movement upon which they are based, are a dangerous technology by which healthcare organizations seek to discipline, govern and regulate nursing work. CONCLUSION(S): Despite the remarkable institutional promotion of 'ready made' and 'ready-to-use' guidelines, we demonstrate how the RNAO deploys BPGs as part of an ideological agenda that is scientifically, socially, politically and ethically unsound. Implications for nursing management Collaborations between health care organizations and professional organizations can become problematic when the latter dictate nursing conduct in such a way that critical thinking is impeded. We believe that nurse managers need to understand that the evidence based movement is the target of well-deserved critiques. These critiques should also be considered before implementing so-called 'Nursing Best Practice Guidelines' in health care milieux. PMID- 18405256 TI - Evidence-informed practice: from individual to context. AB - AIM: This commentary considers the shift in evidence-informed practice away from the individual practitioner to an acknowledgement that context is also important. BACKGROUND: The view of practitioner as 'rational agent' capable of searching, appraising and translating research evidence into individual practice has dominated the literature. However, a growing body of research leads us to question whether evidence use is indeed an individual activity. EVALUATION: Key research studies were purposively selected to build the case for the arguments made. KEY ISSUES: Apart from attitude, there is little to indicate that any potential individual determinants influence research use. Views of what constitutes evidence for evidence-based practice have become more inclusive and sophisticated. Evidence tends to be contextually bound and individually interpreted and particularized within that context. As such, evidence use is beginning to be recognized more widely as a contingent process, which varies across setting and time. A number of contextual factors have been found to be potentially influential including culture and leadership. CONCLUSION(S) AND IMPLICATIONS FOR NURSING MANAGEMENT: It cannot be assumed that evidence-based resources such as clinical guidelines will be accepted at face value by practitioners. Developing the skills of individuals to critically appraise research will not automatically lead to greater evidence use. Reviewing organizations' capacity for evidence-informed practice as a system property and cultural factor may lead to insights about the barriers and facilitators to evidence use. Investing in the capability of key individuals at multiple levels of the organization as leaders of evidence-based practice activities may be one promising organizational strategy. PMID- 18405257 TI - Navigating the evidence-based practice maze. AB - AIM: This article's purpose is to provide nurses with key points to consider in facilitating informed decision making while navigating the evidence-based practice (EBP) maze. BACKGROUND: EBP in nursing evolved from the medical model and continues developing within the holistic nursing paradigm. Inconsistent terminology, multiple applications, and lack of a unifying theory create challenges for nurses. KEY ISSUES: Recognition that multiple knowledge sources to support clinical decision making have merit for facilitating 'best' patient outcomes; EBP must be an internalized value of professional nursing. CONCLUSIONS: Multiple ways of knowing, or evidence, for informed clinical decision making must be considered based on situational context. No hierarchy fits all situations. Nurses must provide support and resources to facilitate nurse empowerment; nurses are accountable for using EBP to enhance patient outcomes. Implications for nursing management Effective EBP implementation relies on nurses being cognizant of what current nursing EBP trends are based on and where they are going. Nurses have a key role in facilitating consensus regarding evidence to be used in EBP, and ensuring availability of resources for empowering nurses to be accountable for outcome-oriented patient care through utilizing EBP. PMID- 18405258 TI - Beyond evidence-based nursing: tools for practice. AB - AIM: This commentary shares my views of evidence-based nursing as a framework for practice, pointing out its limitations and identifying a wider base of appraisal tools required for making good clinical decisions. BACKGROUND: As the principles of evidence-based nursing take an increasingly greater hold on nursing education, policy and management, it is important to consider the range of other decision making tools which are subordinated by this approach. EVALUATION: This article summarizes nursing's simultaneous reliance on and critique of evidence-based practice (EBP) in a context of inadequate critical reasoning. It then provides an exemplar of the limitations of evidence-based practice and offers an alternative view of important precepts of decision-making. KEY ISSUE: I identify means by which nurses can develop skills to engage in informed and robust critique of practices and their underpinning rationale. CONCLUSION: Nurses need to be able to locate and assess useful and reliable information for decision-making. This skill is based on a range of tools which include, but also go beyond EBP including: information literacy, humanities, social sciences, public health, statistics, marketing, ethics and much more. IMPLICATIONS FOR NURSING MANAGEMENT: This essay prompts nursing managers to reflect upon whether a flurried enthusiasm to adopt EBP neglects other important decision-making skills which provide an even stronger foundation for robust nursing decisions. PMID- 18405259 TI - A paradigm for the production of practice-based knowledge. AB - AIM: The aim of this paper was to explore trends, arguments and issues surrounding knowledge production and nursing practice, and to propose a paradigm of practice-based knowledge along with strategies to promote theory-based knowledge development in practice. BACKGROUND: Practice-based knowledge has been marginalized in the current practice and research paradigms. Several reasons for this are presented, some of which may be addressed to facilitate a more inclusive approach to knowledge that can potentially advance patient care and the discipline. EVALUATION: Classic and contemporary scholarly sources in nursing, philosophy, education, social sciences and other areas were critically reviewed and applied to support the argument and propose a paradigm of practice-based knowledge production. KEY ISSUES: A key point identified in the paper is that theory, meaning conceptualizations at all levels of abstraction, is an important tool of knowledge development in nursing practice as it is in traditional research. Conclusions Various strategies exist that can be employed to promote development and use of practice-based knowledge in the clinical setting. The strategies are innovative yet practical, and require the support and encouragement of nursing management for their successful implementation. IMPLICATIONS FOR NURSING MANAGEMENT: Nursing managers can influence if not facilitate all of the strategies to promote practice-based knowledge development identified in the paper. These efforts could give voice to the caregiver's knowledge and, in turn, enhance patient care and the satisfaction and retention of nurses. PMID- 18405260 TI - How patients' experiences contribute to decision making: illustrations from DIPEx (personal experiences of health and illness). AB - AIM: To describe how people use their and other people's experiences ('experiential evidence') in making health care decisions. BACKGROUND: People faced with health decisions may employ not only clinical evidence and advice, but also their own previous experiences and the experiences of others who have faced similar decisions. Professionals have taken little notice of people's use of such experiential evidence and its importance in practice; acknowledging it would improve communication with patients. METHODS: The data come from the DIPEx (personal experiences of health and illness) project, which involves rigorous analysis of narrative interviews of people with particular conditions, chosen to represent the widest practicable range of experiences of each condition. Each collection consists of 40-50 interviews. RESULTS: The analyses of people's experiences are summarized for patients and professionals on a website (http://www.dipex.org) and illustrated by clips (video, audio or written) from the interviews. We draw on the qualitative research conducted for DIPEx to consider some of the different ways that people integrate experiential information when they face decisions about antenatal screening, childhood immunization and treatment for cancer. CONCLUSIONS: Other patients' experiences are an important part of the evidence that people use when making decisions about health care. People are naturally drawn to other people's stories, which add salience to medical information and make facts palatable and memorable. Patients' experiences are not an alternative to the evidence base - they are part of it; to dismiss them as 'anecdotes' is a serious misunderstanding. There are established methods, grounded in social science traditions, which can be used to research and report patients' experiences. Health professionals, as well as patients, need to consider and value this experiential evidence. IMPLICATIONS FOR NURSING MANAGEMENT: Managers and clinicians can help patients by guiding them to information resources, such as DIPEx, that present evidence-based health information through patients' experiences. PMID- 18405261 TI - Tensions and contradictions in nurses' perspectives of evidence-based practice. AB - AIM: To explore nurses' understanding and interpretation of evidence-based practice (EBP). BACKGROUND: EBP has been welcomed into the nursing lexicon without a critical examination of its interpretation by practitioners. The literature suggests that there is a great deal of confusion and contradiction over the meaning and application of EBP. Although work has been conducted on how EBP might be implemented, the general issue of how nurses understand and use EBP is largely unexplored. This paper seeks to examine in depth the understandings of EBP, to enable managers, educationalists and policy makers to implement it more effectively. METHODS: All registered nurses, midwives and health visitors in one UK National Health Service (NHS) Trust were asked to complete a questionnaire in October 2006. RESULTS: Despite a disappointing response rate (8.9%, 218/2438), the survey revealed interesting tensions and contradictions in nurses' understanding of EBP. National and local guidelines, practitioners' own experience and patients' preferences were the main influences on nurses' practice. Published research had relatively little impact, particularly among nurses graded E, F and G and those who had not attended a study day on EBP. CONCLUSIONS: The hierarchies of evidence propounded in local and national guidelines are not adopted by practising nurses, who use other sources of evidence, such as reflection on their own experiences, when making clinical decisions. However, subsuming published evidence to clinical judgement does not contradict the original tenets of EBP. IMPLICATIONS FOR NURSING MANAGEMENT: Unless it is incorporated into national or local guidelines, research has relatively little impact on practice. To develop nursing practice and nursing knowledge, nurse leaders need to foster the synthesis of experiential knowledge and published research, in accordance with the founding principles of the EBP movement. PMID- 18405262 TI - Exploring the evidence base of patient involvement in the management of health care services. AB - AIM: This paper aims to explore the evidence base of patient involvement in the management of health care services. BACKGROUND: It has been suggested that, as well as acknowledging the role that service users could play in deciding their own treatment, they should also be involved in the managerial aspects of health service delivery. However, there is no robust evidence base to support user involvement either in terms of patient preference or in terms of improved service outcomes. METHODS: Seven databases were searched using the search terms 'user involvement' and 'service management'. Terms were adjusted for each database to maximize the number of hits. Each paper was graded using the sophisticated typology developed by the UK Department of Health. RESULTS: Four key issues emerged surrounding organizational power, shared meaning, liaison and user contribution. CONCLUSIONS: There is a reasonable body of robust qualitative evidence surrounding user involvement in health service management but a disappointing lack of quantitative studies or mixed methods approaches. Implications for nursing management Using the emergent messages will assist service mangers in enhancing user/provider collaborations. PMID- 18405263 TI - Clinical geography: nursing practice and the (re)making of institutional space. AB - AIM: To present a geographical study that highlights the wide ranging spatial features of nursing agency. In turn, illustrate the further potential for geographical research to describe, support, challenge and guide clinical practice - particularly with regard to those 'everyday' activities and actions undertaken on a frequent basis. BACKGROUND: To provide a focus, and to anchor the study in existing clinical knowledge and debates, the role of nursing in the (re)making of institutional experiences and life is specifically explored. METHODS: In-depth semi-structured interviews were conducted with 15 nurses working in Buckinghamshire and West London, UK. The following specialties were represented: acute care including emergency (n = 3), midwifery (n = 3), children's nursing (n = 2), elderly care (n = 1), rehabilitation (n = 2), mental health (n = 3) and palliative care (n = 1). RESULTS: Nurses claimed to actively manipulate, normalize and recreate clinical spaces as part of their everyday therapeutic practice. Specifically, the range of agency employed by them falls under the following six categories: adjusting social composition; introducing 'normal' activities; providing private spaces; seeking private spaces; explaining clinical spaces; spaces for personal escape and wellbeing. IMPLICATIONS FOR NURSING MANAGEMENT: It is recommended that nurse leaders - including researchers, managers and clinical educators - explore geography as a source of social scientific evidence that sheds light on the complex nature of everyday professional practice. In this regard, some important disciplinary and structural issues are noted. PMID- 18405264 TI - Interprofessional education and working in mental health: in search of the evidence base. AB - AIM: To explore interprofessional attitudes arising from shared learning in mental health. BACKGROUND: Inter-professional education in healthcare is a priority area for improving team-working and communication. Many studies have attempted to evaluate its benefits and challenges, although few emanate from the mental health arena. However, producing evidence to link educational input with clinical outcomes is notoriously difficult. This project attempted to produce evidence for changes in interprofessional attitudes and stereotypes. METHOD(S): Mental health nursing students and clinical psychology trainees participated in inter-professional education. An evaluation tool was designed to evaluate the experience and outcomes, and to consider implications for interprofessional working. RESULTS: There was an increase in clarity regarding roles, approaches and resources, and how to collaborate in practice. There was no significant change in professional identity. Many challenges were identified, including differences in academic level, previous experience, expectations, assessment, motivation and effort. CONCLUSION: Despite the challenges, it remains important to offer collaboration with future mental health colleagues as a foundation for effective team-working. Recommendations are made for creating inter-professional education opportunities for diverse student groups. IMPLICATIONS FOR NURSING MANAGEMENT: Mental health professionals need to work effectively in multidisciplinary teams. Drawing on available guidance, managers should encourage and support team members to undertake shared learning where possible, both within clinical settings and through more formal educational provision. In this way, managers can facilitate collaborative relationships which will pay dividends for the provision of effective mental health care. This project adds to the limited knowledge currently available on interprofessional learning and attitudes within a mental health context. PMID- 18405265 TI - 'Opposition is true friendship'. Feedback. PMID- 18405267 TI - Association of symptomless TMJ sounds with occlusal force and masticatory performance in older adults. AB - This study investigated associations between temporomandibular joint (TMJ) sounds and occlusal force or masticatory performance stratified by posterior occlusal supports in older Japanese adults. The subjects consisted of 1646 independently living people over 60 years. Masticatory performance, occlusal force, TMJ sounds and maximal mouth opening were examined. Posterior occlusal supports were classified by the Eichner Index. The prevalence of TMJ sounds was 27.7%, limitation of mouth opening (< 40 mm) was 7.9% and TMJ pain was only 1.5%. In the Eichner C group, TMJ sounds were significantly associated with lower occlusal force (OR = 3.20, P = 0.046) and lower masticatory performance (OR = 3.18, P = 0.041) after controlling for gender and age. These associations were not found in the Eichner A and B groups. Within the limitations of this study, the presence of TMJ sounds, even if they were symptomless, was associated with impairment of masticatory function in older adults with reduced occlusal support. PMID- 18405268 TI - Functional status of masticatory system, executive function and episodic memory in older persons. AB - Findings from human experimental studies suggest that mastication positively influences cognitive function. The participants in those studies were relatively young. The goal of this study was to examine the relationship between the functional status of the masticatory system, episodic memory, and executive functions in elderly people. The participants, elderly people living independently at home, were divided into two groups. One group had a full complement of natural teeth (n = 19) and the other group had full dentures (n = 19). The functional status of the masticatory system was assessed by measuring mandibular excursions (i.e. the distances over which the mandible can move in the open, lateral, and forward directions), bite force, number of occluding pairs and complaints of the masticatory system (facial pain, headaches/migraine). Executive functions and episodic memory were assessed by neuropsychological tests. Backward regression analysis showed that only in the group of elderly people with full dentures, 22% of executive functions were predicted by complaints of the masticatory system and 19.4% of episodic memory was predicted by masticatory performance (composed of mandibular excursions and bite force). The conclusion of this study is that only in older persons with full dentures the relationship between mastication, episodic memory, and executive function becomes evident when the functional status of the masticatory system decreases. PMID- 18405269 TI - Masticatory ability and functional tooth units in Japanese adults. AB - The purposes of this study were (i) to examine the relationship between the number of natural teeth and the number of functional tooth units in Japanese adults, (ii) to evaluate how functional tooth units relate to subjective masticatory ability and (iii) to determine the minimum number of natural teeth and functional tooth units needed to maintain adequate self-assessed chewing function. A self-administered questionnaire was given and dental examination was conducted for 2164 residents aged 40 to 75 years. Counts were made on the number of functional tooth units of natural teeth (n-functional tooth units), the sum of natural teeth and artificial teeth on implant-supported and fixed prostheses (nif functional tooth units) and the sum of natural teeth and artificial teeth on implant-supported, fixed and removable prostheses (total-functional tooth units). The average number of natural teeth, n-functional tooth units and nif-functional tooth units decreased with age, but these were often replaced by functional tooth units from artificial teeth on removable prostheses. Total-functional tooth units in 50-59 year old people were slightly lower compared with those in other age groups. Subjects who reported that they could chew every food item on an average had 23.4 total natural teeth, 12.6 posterior natural teeth, 7.6 n-functional tooth units, 8.6 nif-functional tooth units and 10.4 total-functional tooth units, and subjects without chewing difficulties had fewer functional tooth units from removable prostheses. Maintaining 20 and more natural teeth and at least eight nif-functional tooth units is important in reducing the likelihood of self assessed chewing difficulties. PMID- 18405270 TI - Association between food mixing ability and electromyographic activity of jaw closing muscles during chewing of a wax cube. AB - The purpose of this study was to clarify association between food mixing ability and activity of jaw-closing muscles during chewing of a wax cube. Twenty subjects with complete dentitions (mean age 24.1 years) were directed to chew a two coloured paraffin wax cube for 10 strokes on preferred chewing side. Surface electromyograms (EMG) were recorded from the right and left masseter and anterior temporalis muscles during chewing of the wax cube. Maximum voltage, duration and muscle work for burst of each chewing cycle were measured on integrated EMG in each muscle. Food mixing ability was estimated as mixing ability index determined from the colour mixture and shape of the chewed wax cube. Some EMG parameters of all muscles except for masseter muscle on non-chewing side showed significant positive correlations with the mixing ability index (r = 0.45-0.56, P < 0.05). However, most of the EMG parameters correlated with one another. As a result, only muscle work of masseter muscle on the chewing side was identified as a significant predictor accounting for 28% interindividual variation in the mixing ability index (P < 0.01). These results suggest that activity of jaw-closing muscles during chewing the wax cube seems to be weakly related to food mixing ability. PMID- 18405271 TI - Palate morphology of bruxist children with mixed dentition. A pilot study. AB - The objective of the study was to analyse quantitatively palatal morphology in bruxist and non-bruxist children with mixed dentition. Twenty-three children with mixed dentition were classified as bruxist according to their anxiety level, audible occlusal sounds related by the parents and signs of temporomandibular disorders; 23 children were control subjects matched for gender, age, and dental formula. The maxillary dental arches of all subjects were reproduced from alginate impressions cast in dental stone with a standardized technique. The casts were digitalized and mathematical equations were used to obtain the form of the palate in the sagittal, frontal and horizontal planes. Bruxist children had a statistically significant longer palate in the sagittal plane than control children; palatal shape differed especially in correspondence of the third, fourth and fifth teeth, bruxist children showing a relatively higher palate than control children. In this pilot study, sagittal plane differences in the palate between bruxist and non-bruxist children matched for age and gender were found. Further investigations are needed to understand better the clinical implications of the findings. Results should be taken into account in the diagnosis of the occlusal development in children with parafunctions to prevent future abnormalities: a bruxist child may have bigger dental arches than a normal child. PMID- 18405272 TI - The effect of nasal speaking valve on the speech under experimental velopharyngeal incompetence condition. AB - Velopharyngeal incompetence (VPI) is a contributing factor to speech disorders, frequently accompanying disorders, such as cleft palate, congenital paralysis of the soft palate, and cerebrovascular disease. For the treatment of these types of dysarthria, a Nasal Speaking Valve (NSV), which regulates nasal emission utilizing one-way valve, has been reported to be effective. As the unpleasantness while wearing the NSV was less than that with the conventional Palatal Lift Prosthesis, the NSV could be worn for a longer period of time. As NSV is inserted into the nostrils, this device could easily be provided for edentulous patients. This study aimed to evaluate the effect of NSV on experimentally induced VPI condition. Intelligibility of monosyllabic speech, intelligibility of conversational speech, nasalance score and acoustic analysis were used to evaluate the effect of NSV. Local infiltration anaesthesia was achieved to the area of levator veli palatini muscle and tensor veli palatine muscle of seven adult male subjects. In all subjects, remarkable suppression of the soft palate movement could be observed after the local infiltration anesthesia. Although the utterance supported by NSV under the experimentally induced VPI condition was less natural than the normal utterance, the intelligibility was markedly improved by the device. From the results of this study, the acoustic properties of NSV on the pure VPI condition, which was enabled by the newly established experimental induction, were revealed. This experimental model was also proved to provide a basis for the improvement in the treatment modalities for VPI. PMID- 18405273 TI - Relationship between the use of full dentures and mucosal alterations among elderly Brazilians. AB - The objective of the present study was to evaluate the prevalence of oral mucosal lesions associated with the use of full dentures (FD) among non-institutionalized individuals of 60 or more years of age in a rural Brazilian population. The sample consisted of 344 individuals aged 60 or more from two rural communities of Brazil. Of this total, 146 were FD users and 198 FD, non-users. Angular cheilitis, denture stomatitis and inflammatory fibrous hyperplasia were statistically associated with prosthesis use. Hygiene and integrity of the prosthesis were related to the presence of oral lesions. While inflammatory fibrous hyperplasia was positively related to FD integrity, denture stomatitis was associated with time of use, hygiene status and integrity of FD. The results indicate the need for oral health care programmes for the elderly and show a relationship between time of use, quality and hygiene of oral prostheses with the presence of mucosal lesions. PMID- 18405274 TI - Load-bearing capacity of direct four unit provisional composite bridges with fibre reinforcement. AB - The aim of the current in vitro study was to evaluate the load-bearing capacity of provisional four-unit bridges with and without two different types of glass fibre reinforcement produced in a direct technique with a silicon template from the composite materials Protemp, Luxatemp and CronMix. From each composite material, 30 bridges were manufactured, 10 without fibre reinforcement, 10 with Stick/StickNet and 10 with everStick/everStickNet reinforcement. After artificial aging by thermocycling samples were cemented onto a master model and subjected to load testing in a universal testing machine. Mean force at fracture for the groups without fibre reinforcement varied from 486 to 612 N. Depending on the material combination, fibre reinforcement resulted in a slightly increased or a reduced mean load-bearing capacity, varying from 674 N for the combination of Protemp and Stick to 262 N for CronMix and everStick. Position of fibres within the resin pontic was found to have an important influence on load-bearing capacity of reinforced bridges. Scanning electron microscopic investigation showed good adhesion of everStick fibres to the pre-impregnation resin whereas bonding of Stick fibres to the embedding polymer was incomplete. In conclusion, fibre reinforcement with Stick or everStick did not improve the load-bearing capacity of direct provisional bridges made from Protemp, Luxatemp, or CronMix as the position of fibres could not be controlled during the direct manufacturing process. To increase the load-bearing capacity of the provisional bridges, an exact positioning of fibres at the tension side in the dental laboratory is necessary. PMID- 18405275 TI - 3D morphological assessment of occlusal treatment by measuring dental casts with a micro-focus X-ray CT. AB - The analysis of occlusal relationship is important for the success of dental treatment. Three-dimensional (3D) computer models of upper and lower dental casts can play a significant role. In this study, we proposed and applied a new method in actual clinical assessment to measure dental casts with occlusal relationship by using a micro-focus X-ray CT system. We examined the modelling accuracy by comparing multiple 3D images taken by shifting the dental cast position. Modelling accuracy was confirmed as 0.03 mm. One occlusal treatment in clinical practice was selected as a case example. The dental casts and bite impression, taken before treatment, were scanned and the occlusal contacts and distance distribution between the upper and lower casts were visualized by a coloured map and overlaid on the computer models. Distances between the upper and lower casts of selected points were compared before and after the treatment. Initially, the subject had early contact on the anterior teeth, where distance was measured as 0.04 mm, and only one area measured less than 0.15 mm. After treatment, five areas measured less than 0.15 mm. Also, by comparing the dental cast models taken before and after occlusal adjustment of the tooth, the position and amount of adjustment were visualized. We successfully demonstrated the quantitative clinical assessment of occlusal treatment. PMID- 18405276 TI - Is bovine dentine an appropriate substitute for human dentine in erosion/abrasion tests? AB - The study aimed to compare the dentine wear of primary and permanent human and bovine teeth because of erosion/abrasion and evaluate if bovine dentine is an appropriate substitute for human dentine in further erosion/abrasions tests. Dentine samples from deciduous molars and human third molars as well as from calves' and cattle's lower incisors were prepared and baseline surface profiles were recorded. Each day all samples were demineralized in 1% citric acid, tooth brushed with 100 brushing strokes with toothpaste slurry and stored in artificial saliva for the rest of the day. This cycle was run for 20 days. Afterwards, new surface profiles were recorded and dentine wear was calculated by a customized computer program. Dentine wear because of erosion/abrasion was not statistically, significantly different for human third molars and cattle's lower incisors (P = 0.7002). The dentine wear because of erosion/abrasion of deciduous molars and calves' lower incisors was significantly different (P < 0.0000). No statistically significant difference in the dentine wear of human third molars and cattle's lower incisors was observed, so that the use of cattle's lower incisors as substitute for adult human teeth for further investigations in erosion/abrasion studies could be accepted. PMID- 18405278 TI - Letter from Uruguay. PMID- 18405279 TI - 'Break point': a significant moment in the transference. PMID- 18405281 TI - The origins of acts of love: a discussion of 'Break point ' by Stella M. Yardino. PMID- 18405283 TI - On the determination of autistoid organizations in non-autistic adults. AB - Psychogenic autistic phenomena occurring in non-autistic adults are being increasingly discussed in the psychoanalytical literature. The author endeavours to determine autistoid organizations, which are understood to be a particular form of pathological retreat. Together with the use of autistic objects, two dimensional levelling out, the failure of unconscious communication, and of projective identification, a characteristic of these retreats is that they are meant to afford protection against paranoid-schizoid fears of the external and unintegrated states of the internal world. The formation of such autistic reactions should be sought not only in traumatic separations and the inadequate containment of primary objects, but also in constitutional weakening. Intolerance accompanying projective identification and damage to primal fantasies/preconceptions are discussed. In conclusion, a clinical illustration with autistoid and hypochondriacal retreats clarifies some of the aspects discussed. PMID- 18405284 TI - Immersion versus interactivity and analytic field. AB - Losing oneself in a story, a film or a picture is nothing but another step in the suspension of disbelief that permits one to become immersed in the 'novel' of reality. It is not by chance that the text-world metaphor informs classical aesthetics that, more than anything else, emphasizes emotional involvement. On the contrary, as in much of modern art, self-reflexivity and metafictional attention to the rhetoric of the real, to the framework, to the conventions and to the processes of meaning production, all involve a disenchanted, detached and sceptic vision--in short, an aesthetics of the text as game. By analogy, any analytic style or model that aims to produce a transformative experience must satisfactorily resolve the conflict between immersion (the analyst's emotional participation and sticking to the dreamlike or fictional climate of the session, dreaming knowing it's a dream) and interactivity (for the most part, interpretation as an anti-immersive device that 'wakes' one from fiction and demystifies consciousness). In analytic field theory the setting can be defined- because of the weight given to performativity of language, to the sensory matrix of the transference and the transparency of the medium--the place where an ideal balance is sought between immersion and interaction. PMID- 18405285 TI - The destruction of time in pathological narcissism. AB - This paper describes the characteristics of subjective time (in contrast to objective time), with particular reference to a specific form of pathological experience and relation to the passage of time in patients with narcissistic personality undergoing psychoanalytic treatment. The clinical manifestations and technical approach to this pathology of time experience are outlined in the context of illustrative clinical vignettes. PMID- 18405286 TI - Ethics in the termination of analysis. AB - The author argues that the termination of analysis raises not only theoretical and technical issues but also problems of evaluation that are both moral (what is good or bad with regard to termination) and ethical (what is the best way of achieving the intended objective). Broadly speaking, he considers that the subject can be addressed from three different aspects: (a) a psychoanalysis of ethics; (b) an ethic of psychoanalytic treatment; (c) an area of intersection between psychoanalysis and ethics that has to do with mankind 's norms and values. The termination is particularly well suited to investigation of the intertwining of ethics with other aspects of psychic functioning. A specific description is given of the conflict between the 'limit' and 'completeness', the limit being connected with the analysis and the wish, while completeness is the possibility of experiencing the analysis and one's personal life as endowed with the 'sense of an end-point'. The conflict may be expressed in dramatic or tragic forms that can be productively explored through the Aristotelian concepts of peripeteia and recognition. The termination process offers material for establishing an ethics of the limit, highlighting the psychic role of moral judgement and the need to evaluate this role if a satisfactory termination is to be achieved. PMID- 18405287 TI - Trauma and traumatic neurosis: Freud's concepts revisited. AB - The authors examine Freud 's concepts of 'trauma', 'protective shield against stimuli ' and 'traumatic neurosis' in the light of recent findings. 'Protective shield against stimuli' is regarded as a biological concept which appears in mental life as the striving to avoid unpleasant affects. 'Trauma' is a twofold concept in that it relates to mental experience and links an external event with the specific after-effects on an individual 's psychic reality. A distinction needs to be made between mentally destructive trauma and affective trauma. A destructive trauma does not break through the protective shield but does breach the pleasure-unpleasure principle, so that in the course of its subsequent mastery it leads to a traumatic neurosis. An affective trauma can be warded off under the rule of the pleasure-unpleasure principle and leads to a psychoneurosis. PMID- 18405288 TI - Employing multiple theories and evoking new ideas: the use of clinical material. AB - In this paper, I wish to illustrate how working with a patient who had a certain kind of narcissistic difficulty led me to develop particular clinical strategies to facilitate the development of a sturdier sense of self, greater affect tolerance and modulation, the diminution of harshness of her superego, and the ownership of projected parts of herself, and to decrease paranoid ideation. I call upon concepts from various theoretical schools of psychoanalysis to make sense of the dynamic intricacies of the patient's psychological organization as they revealed themselves in the analytic process. These conceptualizations of the patient's difficulties and of clinical interventions to address them result in a hybrid theory of both theory and technique. What transpired in the clinical work also led me to propose an additional way to understand this kind of patient's difficulties with accepting interpretations or any view that differed from the patient's subjectivity. I am proposing that 'otherness' itself, rather than only specific conflictual aspects of the self, is disowned. It is the analyst's empathic stance toward all that is repudiated--the specific disowned aspects of the self and 'otherness' itself--along with empathy for the patient's conscious state that will enable reinternalization and ultimately healing. PMID- 18405289 TI - History of the unconscious in Soviet Russia: from its origins to the fall of the Soviet Union. AB - Russia accepted the notion of the unconscious and psychoanalysis before many Western countries. The first Russian Psychoanalytic Society was established in 1911. After World War I and the Russian Revolution, for a short happy period, the following psychoanalysts were active: Sabina Spielrein, Tatiana Rosenthal, Moshe Wulff, Nikolai Osipov and Ivan Ermakov. Scholars associated with Soviet ideas participated too, including Aleksandr Luria, Michail Rejsner and Pavel Blonskij. Lev Vygotskij himself dealt with the unconscious. A second psychoanalytical society was set up in Kazan. Unfortunately, at the end of the 1920s, repression dissolved the psychoanalytic movement. Even the word 'psychoanalysis' was banned for decades. Nonetheless, interest in the unconscious, as distinct from psychoanalytic theory, survived in the work of the Georgian leader D. Uznadze. His followers organized the 1979 International Symposium on the Unconscious, in Tbilisi, Georgia, which marked the breaking of an ideological barrier. Since then, many medical, psychological, philosophical and sociological scholars have taken an interest in the unconscious, a subject both feared, for its ideological implications, and desired. Since the 1980s, psychoanalytic ideas have been published in the scientific press and have spread in society. The fall of the USSR in 1991 liberalized the scientific and institutional development of psychoanalysis. PMID- 18405290 TI - Phantastic objects and the financial market's sense of reality: a psychoanalytic contribution to the understanding of stock market instability. AB - This paper sets out to explore if standard psychoanalytic thinking based on clinical experience can illuminate instability in financial markets and its widespread human consequences. Buying, holding or selling financial assets in conditions of inherent uncertainty and ambiguity, it is argued, necessarily implies an ambivalent emotional and phantasy relationship to them. Based on the evidence of historical accounts, supplemented by some interviewing, the authors suggest a psychoanalytic approach focusing on unconscious phantasy relationships, states of mind, and unconscious group functioning can explain some outstanding questions about financial bubbles which cannot be explained with mainstream economic theories. The authors also suggest some institutional features of financial markets which may ordinarily increase or decrease the likelihood that financial decisions result from splitting off those thoughts which give rise to painful emotions. Splitting would increase the future risk of financial instability and in this respect the theory with which economic agents in such markets approach their work is important. An interdisciplinary theory recognizing and making possible the integration of emotional experience may be more useful to economic agents than the present mainstream theories which contrast rational and irrational decision-making and model them as making consistent decisions on the basis of reasoning alone. PMID- 18405291 TI - Panel report: emigration from Berlin. Part One: Transfer of theories and institutional regulations. PMID- 18405292 TI - Panel report: emigration from Berlin. Part Two: Transfer of theories and institutional regulations. PMID- 18405293 TI - Panel report: emigration from Berlin. Part Three: Emigration of education and theory to North America. PMID- 18405294 TI - On projective identification: back to the beginning. PMID- 18405296 TI - David Cronenberg's Spider: between confusion and fragmentation. PMID- 18405297 TI - The Truman Show: how's it going to end? PMID- 18405307 TI - Abstracts of the XXIst International Symposium on Technological Innovations in Laboratory Hematology. April 28-May 1, 2008. Sydney, Australia. PMID- 18405308 TI - NK cell tolerance and the maternal-fetal interface. AB - Natural killer (NK) cells play a fundamental role in the innate immune response through their ability to secrete cytokines and kill target cells without prior sensitization. These effector functions are central to NK cell anti-viral and anti-tumor abilities. Due to their cytotoxic nature, it is vital that NK cells have the capacity to recognize normal self-tissue and thus prevent their destruction. In addition to their role in host defense, NK cells accumulate at the maternal-fetal interface and are thought to play a critical role during pregnancy. The close proximity of uterine NK (uNK) cells to fetal trophoblast cells of the placenta would seemingly lead to catastrophic consequences, as the trophoblast cells are semi-allogeneic. A fundamental enigma of pregnancy is that the fetal cells constitute an allograft but, in normal pregnancies, they are in effect not perceived as foreign and are not rejected by the maternal immune system. Although the mechanisms involved in achieving NK cell tolerance are becoming increasingly well-defined, further clarification is required, given the clinical implications of this work in the areas of infection, transplantation, cancer and pregnancy. Herein, we discuss several mechanisms of NK cell tolerance and speculate as to how they may apply to uNK cells at the maternal-fetal interface. PMID- 18405309 TI - Clinical implication of natural killer cells and reproduction. AB - The regulation of natural killer (NK) cells in the peripheral blood and endometrial layers has been associated with reproductive immunopathology such as recurrent spontaneous abortions (RSA), infertility of implantation failures, or pre-eclampsia. The placenta has a complex anatomical structure and different subsets of NK cells with various functional roles can directly interact with trophoblasts. NK cell subpopulations and their functions, putative roles of NK cells in peripheral blood and endometrium are reviewed in relation to RSA and infertility. An increase in NK cell numbers and /or activity in pre- or post conceptional period in women with RSA or infertility with multiple implantation failures are a significant clinical concern. In addition, immuno-phenotypic characteristics of NK cells in these women support the changes for their increased activity status. Further studies are needed to explore underlying mechanism of NK cells in RSA, infertility, and other reproductive immunopathologies. Possible neurological and hormonal control of NK cells and NK cell interaction with various leukocyte populations need further investigation in women with reproductive failures. PMID- 18405310 TI - Human NK cells in pregnant uterus: why there? AB - Human Natural Killer (NK) cells are present in great number in pregnant uterine mucosa. They must be there for specialized functions, but which ones? This review discusses important recent observations that further contribute to this fascinating debate. Firstly, an array of corroborating findings indicates that uterine NK cell proliferation is synchronized with the cyclic surge of progesterone. Secondly, uterine NK cells are unlikely to exert a direct control on the embryo implantation. Thirdly, these NK cells influence the uterine vascular remodeling in early pregnancy but might not be the single key element that control trophoblast invasion. Finally, uterine NK cells are likely to be an important component of the local maternal immune response to pathogen infections. PMID- 18405311 TI - Secreted heat shock protein gp96-Ig: an innovative vaccine approach. AB - Heat shock proteins (HSPs) are a large family of proteins with different molecular weights and different intracellular localizations. These proteins undertake crucial functions in maintaining cell homeostasis, and therefore they have been conserved during evolution. HSP gp96 also known as glucose-regulated protein grp94, is the primary chaperone of the endoplasmatic reticulum. Gp96/grp94, because of its peptide chaperone capacity and its ability to interact actively with professional antigen-presenting cells (APCs), is also endowed with crucial immunological functions such as natural adjuvant for priming innate and adaptive immunity. To make gp96 accessible to the immune system without biochemical purification and without cell lysis, we generated a secreted form of gp96. The immunological properties of secreted gp96 and its implications for vaccine in human cancer and infectious diseases will be discussed. PMID- 18405312 TI - Mechanisms underlying recruitment and accumulation of decidual NK cells in uterus during pregnancy. AB - Natural killer (NK) cells represent the most prominent immune cell type found in the uterus in the first trimester of human pregnancy and in the secretory phase of menstrual cycle. The role of NK cells in pregnancy has been largely discussed over the past years and it is now becoming increasingly clear that they may influence pregnancy outcome at several levels. In normal pregnancy, it appears that the major function of NK cells is to provide benefit by secreting a number of cytokines, chemokines and angiogenic factors rather than to exert a cytotoxic activity. However, the origin of decidual NK cells is still debated and it remains unclear whether they can derive from NK cell populations recruited from peripheral blood and/or other tissues or from self renewal of NK cell progenitors present in the uterus prior to pregnancy or recruited from other tissues. Here, we review the molecular mechanisms underlying peripheral blood NK cell recruitment and its role in the accumulation of NK cells in the decidua during early pregnancy. PMID- 18405313 TI - Evolution of non-cytotoxic uterine natural killer cells. AB - The immune tolerance and de novo vascularization are two highly intriguing processes at the maternal-fetal interface that appear to be central to normal pregnancy outcome. Immune tolerance occurs despite the local presence of an active maternal immune system including macrophages, dendritic cells and specialized CD56(bright)CD16(-) uterine natural killer (uNK) cells (65-70%). Recent observations indicate that the phenotypic and functional repertoire of uNK cells is distinct from peripheral blood NK and endometrial NK cells, challenging the understanding of their temporal occurrence and function. Origin and specialized programming of uNK cells continue to be debated. uNK cells, replete with an armamentarium to kill the foreign, tolerate the conceptus and facilitate pregnancy. Why do these uNK cells remain non-cytotoxic? Are these NK cells 'multitasking' in nature harboring beneficial and detrimental roles in pregnancy? Are there distinct subpopulations of NK cells that may populate the decidua? We propose that the endometrium/decidua functions as an 'inducible tertiary lymphoid tissue' that supports the recruitment and expansion of CD56(bright)CD16(-) NK cells and induces transcriptional up-regulation of angiogenic machinery in response to exposure to local hormonal factors, cytokine milieu and perhaps hypoxia. The angiogenic features of uNK cells could further result in a 'multitasking' phenotype that still remains to be characterized. This article discusses the factors and pathways that bridge the angiogenic and non-cytotoxic response machineries at the maternal-fetal interface. PMID- 18405316 TI - Attend the 9th World Congress of Bioethics! PMID- 18405314 TI - Decidual natural killer cell tuning by autologous dendritic cells. AB - PROBLEM: Dendritic cells (DC)/natural killer (NK) cells interactions in the deciduas of early human pregnancies were analyzed in vitro. METHOD OF STUDY: Phenotype, cytokine expression and/or cytolytic mediators' expression were measured by flow cytometry in NK and DC from the freshly isolated decidual mononuclear cells or after their purification and co-culture in vitro. Proliferation of 5(6)-Carboxyfluorescein diacetate N-succinimidyl ester (CFSE) labeled CD56(+) cells was analyzed by flow cytometry after the co-culture with CD1a(+) or CD83(+) DC. RESULTS: Decidual CD1a(+) cells show less mature phenotype with no expression of CD197, lower expression of CD80 and CD86 and higher expression of CD206 and CD195 in comparison to CD83(+) cells. Interleukin (IL) 15, interferon-gamma and tumor necrosis factor-alpha productions were higher in immature than mature DC, whereas IL-10 and IL-18 were equally produced in both subpopulations. Immature DC increase perforin, FasL and TRAIL protein expression and proliferation of NK cells, but decrease their intracellular IL-15 production. Mature DC caused less efficient proliferation of NK cells, and did not affect cytokine and cytolytic mediator expression. CONCLUSION: These results suggest that decidual CD1a(+) cells regulate and shape NK cell function more profoundly than CD83(+) cells in decidua. PMID- 18405317 TI - Vulnerability in research and health care; describing the elephant in the room? AB - Despite broad agreement that the vulnerable have a claim to special protection, defining vulnerable persons or populations has proved more difficult than we would like. This is a theoretical as well as a practical problem, as it hinders both convincing justifications for this claim and the practical application of required protections. In this paper, I review consent-based, harm-based, and comprehensive definitions of vulnerability in healthcare and research with human subjects. Although current definitions are subject to critique, their underlying assumptions may be complementary. I propose that we should define vulnerability in research and healthcare as an identifiably increased likelihood of incurring additional or greater wrong. In order to identify the vulnerable, as well as the type of protection that they need, this definition requires that we start from the sorts of wrongs likely to occur and from identifiable increments in the likelihood, or to the likely degree, that these wrongs will occur. It is limited but appropriately so, as it only applies to special protection, not to any protection to which we have a valid claim. Using this definition would clarify that the normative force of claims for special protection does not rest with vulnerability itself, but with pre-existing claims when these are more likely to be denied. Such a clarification could help those who carry responsibility for the protection of vulnerable populations, such as Institutional Review Boards, to define the sort of protection required in a more targeted and effective manner. PMID- 18405318 TI - What should research participants understand to understand they are participants in research? AB - To give valid informed consent to participate in clinical research, potential participants should understand the risks, potential benefits, procedures, and alternatives. Potential participants also should understand that they are being invited to participate in research. Yet it is unclear what potential participants need to understand to satisfy this particular requirement. As a result, it is unclear what additional information investigators should disclose about the research; and it is also unclear when failures of understanding in this respect undermine the validity of potential participants' informed consent. An analysis of individuals' interests suggests that potential participants need to understand three additional facts to understand that they are being invited to participate in research: 1) research contribution: those who enroll in the study will be contributing to a project designed to gather generalizable knowledge to benefit others in the future; 2) research relationship: the investigators will rely on participants' efforts to gather the generalizable knowledge to benefit others; and 3) research impact: the extent to which participating in the study will alter what participants do and what happens to them. PMID- 18405319 TI - Environmental health research on hazards in the home and the duty to warn. AB - When environmental health researchers study hazards in the home, they often discover information that may be relevant to protecting the health and safety of the research subjects and occupants. This article describes the ethical and legal basis for a duty to warn research subjects and occupants about hazards in the home and explores the extent of this duty. Investigators should inform research subjects and occupants about the results of tests conducted as part of the research protocol only if the information is likely to be accurate, reliable, and medically useful. Investigators should warn subjects and occupants about hazards they happen to discover while they are in the home, if a reasonable person would warn the subjects and occupants about those hazards. Investigators should not report illegal hazards discovered in the home to the authorities, unless those hazards constitute abuse or neglect of children or mentally disabled people living in the home. When investigators decide to warn research subjects and occupants about hazards in the home, they should take some steps to help them make effective use of this information, such as providing additional counselling or making a referral for remediation or medical treatment. Investigators should discuss these issues with research subjects during the informed consent process. PMID- 18405320 TI - Moral qualms, future persons, and embryo research. AB - Many people have moral qualms about embryo research, feeling that embryos must deserve some kind of protection, if not so much as is afforded to persons. This paper will show that these qualms serve to camouflage motives that are really prudential, at the cost of also obscuring the real ethical issues at play in the debate concerning embryo research and therapeutic cloning. This in turn leads to fallacious use of the Actions/Omissions Distinction and ultimately neglects the duties that we have towards future persons. PMID- 18405321 TI - Research involving prisoners: consensus and controversies in international and European regulations. AB - This article examines international and European regulations on research involving prisoners for consensus, differences, and their consequences, and offers a critical evaluation of the various approaches. Agreement exists that prisoners are at risk of coercion, which might interfere with their ability to provide voluntary informed consent to research. Controversy exists about the magnitude of this risk and the consequences that should follow from this risk. Two strategies are proposed for a method of protecting prisoners that does not lead to discrimination: first, more caution to assure non-coerced consent and second, restrictions on the type of research. Most regulations stress the importance of the principle of equivalence of healthcare in places of detention as part of an efficient protection against research risks and discrimination. All the presented approaches have shortcomings. While 'over-use' of prisoners for research as compared to the general population is ethically unjustified, not granting prisoners access to studies beneficial to their own health because of over-strict regulations is equally unjustified. A middle solution should be preferred, one that grants a minimum of protection together with the lowest possible barriers. Research that does not entail a direct benefit for the individual detainee should be restricted to types of research that have a benefit for detainees as a group and that are of low risk. What will ultimately protect prisoners best, while producing the greatest benefit for them, is access to the same healthcare available to members of the community including research as a true option. PMID- 18405322 TI - 'Fair benefits' accounts of exploitation require a normative principle of fairness: response to Gbadegesin and Wendler, and Emanuel et al. AB - In 2004 Emanuel et al. published an influential account of exploitation in international research, which has become known as the 'fair benefits account'. In this paper I argue that the thin definition of fairness presented by Emanuel et al, and subsequently endorsed by Gbadegesin and Wendler, does not provide a notion of fairness that is adequately robust to support a fair benefits account of exploitation. The authors present a procedural notion of fairness--the fair distribution of the benefits of research is to be determined on a case-by-case basis by the parties involved in each study. The fairness of the distribution of benefits is not assessed against an independent normative standard. Emanuel et al.'s account of fairness provides a framework for objecting only to transactions that occur without the fully informed consent of the weaker party. As a result, a debate about exploitation collapses into a debate about consent. This is problematic because, as the proponents of the fair benefits framework acknowledge, neither the trial participants' consent nor the host community's consent preclude exploitation. Attempts to stipulate normative standards of fairness to protect research subjects in developing countries have been controversial and divisive, and it is therefore understandable that bioethicists would be tempted to develop accounts of exploitation that are independent of such prescriptive principles. I conclude, however, that the utility of the fair benefits model of exploitation as a policy tool will ultimately depend on whether a substantive principle of fairness can be developed to underpin it. PMID- 18405324 TI - Cerebral gene expression of superantigen encephalitis in the lewis rat induced by staphylococcal enterotoxin a. AB - Superantigens were suggested to play a role in the pathogenesis of different autoimmune diseases including multiple sclerosis (MS). Previously, it was demonstrated that local expression of the superantigen, staphylococcal enterotoxin A (SEA) in the brain of rats may lead to encephalitis which was amplified by using intravenous injection of concanavalin A (ConA)-activated splenocytes. In the present investigation, gene expression was studied in the rat brain 8 days after an injection of 50 mul of 1 mg/ml SEA or saline and 5 days after an intravenous injection of 1 x 10(7) ConA-activated spleen cells. Of 8800 genes investigated (Affymetrix, rat genome U34A), the expression of 106 genes was significantly and at least threefold increased with SEA, while the expression of 29 genes was decreased at least threefold. Increased gene expression was compatible with an intracerebral inflammatory response mediated by antigen presenting cells and CD8+ T lymphocytes. Elevated chemokines comprised RANTES (CCL5), osteopontin, MCP-1 (CCL2) and CXCL10. Further, genes with increased expression were assigned to the extracellular matrix, microglia/macrophage cell elements, astrocytes (GFAP) and phagocytosis. There was considerable conformity between previously reported gene expression profiles for experimental autoimmune encephalomyelitis (EAE) or MS and the present findings. Our data are in line with the concept that T-cell superantigen locally expressed in the central nervous system induces an inflammatory response. Therefore, the study of gene expression profiles does not seem to allow clear conclusions with respect to the aetiology of central nervous system autoimmune diseases. PMID- 18405323 TI - Expression of CD68 in non-myeloid cell types. AB - CD68, the human homologue of macrosialin, is commonly regarded as a selective marker for human monocytes and macrophages. Its expression is thought to be regulated by a macrophage-specific promoter. However, several immunohistochemical studies have indicated that CD68 antibodies also react with other haematopoietic and non-haematopoietic cell types. We investigated the expression of CD68 in various primary cells and carcinoma cell lines using immunohistochemistry, flow cytometry, Western blot analysis and qRT-PCR. Weak but significant immunoreactivity was detected in lymphocytes and several tumour cell lines whereas staining of primary fibroblasts and endothelial cells was comparable to macrophages. The intensity of CD68 staining in individual cell types depended on the antibody clone and the fixation technique. Anti-CD68 mAb KP1 should be used with great caution for frozen tissue sections due to its reactivity with a wide variety of cell types. Also, care should be taken when distinguishing macrophages from fibroblasts/stromal cells in paraffin sections after formalin fixation since both cell types are stained highly positive for CD68. In accordance, mRNA expression of CD68 was not only detected in macrophages and monocytes but also in fibroblasts as well as endothelial cells and tumour cells, although with a varying intensity. Cloning of full length 5'-sequences and determination of transcription start sites shows that macrophages and fibroblasts initiate transcription within the known promoter region; however, from different start sites, indicating alternative promoter architecture in myeloid versus non-myeloid cells. We suggest that CD68 is not a selective macrophage marker but rather a lysosomal protein that is enriched in macrophages. PMID- 18405325 TI - Toll-like receptor cross-hyporesponsiveness is functional in interleukin-1 receptor-associated kinase-1 (IRAK-1)-deficient macrophages: differential role played by IRAK-1 in regulation of tumour necrosis factor and interleukin-10 production. AB - Signalling downstream Toll-like receptors (TLR) is regulated at several levels in order to activate the immune response and prevent excessive inflammation. Altered intracellular signalling may be one reason that repeated stimulation of various TLRs results in hyporesponsiveness and cross-tolerance. We report that TLR cross tolerance is inducible in the absence of interleukin-1 receptor-associated kinase 1 (IRAK-1) in peritoneal macrophages. Similar to wild-type macrophages, IRAK-1 deficient macrophages respond with decreased tumour necrosis factor (TNF) production to a secondary TLR stimulation, but in opposite to IRAK-1(+/+), IRAK 1(-/-) macrophages display increased interleukin (IL)-10 production at TLR restimulation. IRAK-1-deficient peritoneal macrophages have a defective TNF and IL-10 production in response to lipoteichoic acid stimulation as well as a defective IL-10-but a normal TNF production in response to high concentration of lipopolysaccharide. Our results demonstrate that IRAK-1 is not necessary for induction of TLR cross-tolerance as judged by TNF production. PMID- 18405326 TI - Prolonged eosinophil production after allergen exposure in IFN-gammaR KO mice is IL-5 dependent. AB - Asthma is a T helper 2 (Th2)-driven inflammatory process characterized by eosinophilia. Prolonged airway eosinophilia is commonly observed in asthma exacerbations. Our aim was to evaluate whether eosinophilia in prolonged allergic inflammation is associated with a continuous supply of new eosinophils to the airways, and how this is regulated. Ovalbumin (OVA)-sensitized interferon-gamma receptor knockout mice (IFN-gammaR KO), known to maintain a long-lasting eosinophilia after allergen exposure, were compared to wild type (wt) controls. Animals were exposed to OVA or phosphate-buffered saline on three consecutive days, and bone marrow (BM), blood and bronchoalveolar lavage (BAL) samples were collected 24 h, 7 and 21 days later. Newly produced cells were labelled using bromodeoxyuridine (BrdU). Serum IL-5 was measured and its role was investigated by administration of a neutralizing anti-IL-5 antibody. In-vitro eosinophilopoiesis was examined in both groups by a colony-forming assay. Allergen challenge increased eosinophils in BM, blood and BAL, in both IFN-gammaR KO and wt mice, both 24 h and 7 days after the last allergen exposure. At 21 days after the last exposure, only IFN-gammaR KO mice maintained significantly increased eosinophil numbers. Approximately 50% of BAL granulocytes in IFN-gammaR KO were produced during the last 6 days. Interleukin (IL)-5 concentration was increased in IFN-gammaR KO mice, and anti-IL-5 reduced eosinophil numbers in all compartments. Increased numbers of eosinophil colonies were observed in IFN gammaR KO mice after allergen exposure versus controls. In this model of a Th2 driven prolonged allergic eosinophilia, new eosinophils contribute to the extended inflammation in the airways by enhanced BM eosinophilopoiesis in an IL-5 dependent manner. PMID- 18405327 TI - CD8alpha+beta(low) effector T cells in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a complex autoimmune disorder characterized by the loss of self-tolerance to nuclear antigens. Aberrant T-cell function plays a central role in lupus pathogenesis. We and others previously demonstrated that peripheral TCRalphabeta+CD3+ T cells express CD8beta either at a high (CD8beta(high)) or low density (CD8beta(low)), thereby defining two functionally distinct subsets. CD8beta(low) T cells express predominantly CD8alphaalpha and less CD8alphabeta as a coreceptor, display a differentiated phenotype and exert effector function. CD8beta(high) T cells appear to be the precursors expressing predominantly the heterodimeric efficient CD8alphabeta coreceptor, exhibiting a naive phenotype and high proliferative capacity. In the present study, the distribution and functional properties of CD8beta(high) and CD8beta(low) T cells of SLE patients were compared (n = 20) with those of healthy subjects (n = 16). It was found that expansion of CD8beta(low) T-cell subset correlated with disease activity indicating chronic antigenic stimulation leading to a major lack of naive CD8beta(high) precursor T cells in SLE. Functional characteristics of CD8beta(low) T cells including production of cytokines and cytotoxic granules were not significantly different between patients with SLE and healthy individuals. We speculate that unbalanced CD8beta(high)/CD8beta(low) T-cell relation reflects a skewed homeostasis within the CD8+ T-cell compartment towards fully differentiated effector T cells possibly due to persistent antigen stimulation in SLE. PMID- 18405328 TI - Autoimmune markers in lymphoid malignancies. AB - Chronic immune stimulation such as Helicobacter pylori (hp) infection, Sjogren's syndrome or coeliac disease may initiate non-Hodgkin lymphoma (NHL). The opposite (appearance of autoimmunity) has also been reported. The aim of this study was to describe the pattern of these immune markers in patients with lymphoid malignancies. Sera from 96 patients with NHL (median age 72, range 38-88, F/M 41/55) were analysed with ELISA to determine the frequency of antibodies against guinea pig (gp) and human recombinant (hr) transglutaminase type 2 (Tg2), and hr factor XIII subunit a* (part of the Tg-family), extractable nuclear antigen (ENA), and hp. As hp antibodies decrease in younger age cohorts a sex- and age matched control group of 768 persons was used. The control population for transglutaminase antibodies consisted of 59 blood donors, (median 42 years, range 19-65) was analysed with a commercial kit. Gp-Tg2-IgG positivity was documented in 72% and hr-Tg2-IgG positivity in 15% (5% positive controls for both; P < 0.001 and ns, respectively). For IgA 3% had gp-Tg2 and 4% hr-Tg2 (5% in controls: ns for both). Anti-FXIII-IgA positivity was found in 22% (5% in controls; P = 0.03). Unspecific anti-ENA-IgG positivity was found in 24% (P < 0.001), while only 2% had specific ENA autoantibodies. Moreover, 36% were positive for anti-hp-IgG, while controls were positive in 54% (P < 0.001). The frequency of unspecific autoantibodies was increased. No differences could be noted in specific autoantibodies (hr-Tg2-IgA). In contrast, fewer than expected were anti-hp positive. A defective immune response, similar to that in autoimmune diseases, could contribute to the pathogenesis of lymphoid malignancies. PMID- 18405329 TI - CCR5 Delta32 polymorphism: associated with gallbladder cancer susceptibility. AB - Inflammation of gallbladder is an established risk factor for gallbladder cancer (GBC) pathogenesis. Chemokine receptors play crucial role in antitumour immunity and are involved in inflammation and pathogenesis of cancers. Present study was aimed to examine the role of CCR5 Delta32 polymorphism in conferring genetic susceptibility to GBC. Present case-control study included 144 proven GBC patients and 210 healthy controls. Genotyping was done by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Statistically significant difference was observed in distribution of CCR5+/Delta32 genotype (P = 0.028) [odds ratio (OR) = 2.850; 95% confidence interval (CI) = 1.1-7.2] and CCR5 Delta32 allele (P = 0.012) (OR = 3.145, 95% CI = 1.2-7.7) in GBC patients which was conferring high risk. Stratification of GBC patients showed significant association of CCR5+/Delta32 genotype and CCR5 Delta32 allele with GBC patients with and without gallstones. Analysis based on age of onset and gender suggested significant association of CCR5 Delta32 allele with early onset (<50 years) of the disease but only marginal influence of gender in CCR5 Delta32-mediated risk of cancer. Risk was further modulated by tobacco usage and significantly increased risk was observed in tobacco users with CCR5+/Delta32 genotype. In conclusion, CCR5+/Delta32 genotype and CCR5 Delta32 allele confer significant risk for GBC particularly in patients with early onset and tobacco usage. Role of CCR5+/Delta32 polymorphism in GBC susceptibility is independent of gallstone formation. PMID- 18405330 TI - Inorganic carbon utilisation by freshwater and marine photosynthetic microorganisms. PMID- 18405331 TI - Insights into the evolution of CCMs from comparisons with other resource acquisition and assimilation processes. AB - Regarding inorganic carbon as 'just another' chemical resource used in the growth of aquatic photolithotrophs, we ask three questions and then attempt to answer them. (1) How common are catalysed chemical changes of the resource outside the cell, and accumulation of the resource inside the cell prior to assimilation, for the diverse chemical resources used? (2) Do acquisition and assimilation meet evolutionary optimality criteria with respect to the use of other resources? (3) Are there clues to the evolutionary origin of inorganic carbon concentrating mechanism (CCMs) in the mechanisms of acquisition of other resources and vice versa? Evidence considered includes molecular genetic similarities between CCM components and components of other resource acquisition mechanisms, and palaeogeochemical evidence on the timing of restrictions on the availability of the resources such that extracellular transformation of materials, and their accumulation within cells prior to assimilation, are needed. Provisional answers to the questions are as follows: (1) Many common chemical resources other than inorganic carbon are subject to extracellular chemical conversion and/or accumulation prior to assimilation, e.g. ammonium, nitrate, urea, amino acids, organic and inorganic phosphate and iron; (2) There is some evidence for optimality of CCMs and of less complex resource acquisition processes, exemplified by NH(4)(+) entry and assimilation, though many more data are needed and (3) There are molecular genetic similarities between CCM components and transporters for other solutes and components of respiratory NADH dehydrogenases that are consistent with their use in CCMs representing a derived evolutionary state. Palaeogeochemical evidence suggests that CCMs evolved later than did at least some of the extracellular chemical transformation and/or accumulation mechanisms for other resources. PMID- 18405332 TI - Identification and characterization of two closely related beta-carbonic anhydrases from Chlamydomonas reinhardtii. AB - Aquatic photosynthetic organisms such as the green alga Chlamydomonas reinhardtii respond to low-CO(2) conditions by inducing a CO(2) concentrating mechanism (CCM). Important components of the CCM are the carbonic anhydrases (CAs), zinc metalloenzymes that catalyze the interconversion of CO(2) and HCO(-)(3). Six CAs have previously been identified in C. reinhardtii. Here, we identify and characterize two additional beta-type CAs. These two CAs are closely related beta type CAs and have been designated as CAH7 and CAH8. Conceptual translation shows that CAH7 and CAH8 encode proteins of 399 and 333 amino acids, respectively, and they contain targeting sequences. An unusual characteristic of these two CAs is that they have carboxy-terminal extensions containing a hydrophobic sequence. Both these CAs are constitutively expressed at the transcript and protein level. The CAH7 and CAH8 open reading frames were cloned in the overexpression vector pMal-c2x and expressed as recombinant proteins. Activity assays showed that CAH7 and CAH8 are both active CAs. Antibodies were raised against both CAH7 and CAH8, and immunolocalization studies showed that CAH8 was localized in the periplasmic space. A possible role for CAH8 in the inorganic carbon acquisition by C. reinhardtii is discussed. PMID- 18405333 TI - Diversity of inorganic carbon acquisition mechanisms by intact microbial mats of Microcoleus chthonoplastes (Cyanobacteriae, Oscillatoriaceae). AB - The dissolved inorganic carbon (DIC) acquisition mechanisms were researched in intact microbial mats dominated by the cyanobacteria Microcoleus chthonoplastes Thuret, by determining the effect on photosynthesis of different inhibitors. The microbial mats exhibited high affinity for DIC at alkaline pH, with K(m(DIC)) values similar to the ones described for pure cultures of cyanobacteria and algae in which carbon concentrating mechanisms have been researched. Besides, the photosynthesis was non-sensitive to pH changes within the range of 5.6-9.6, indicating that HCO(3)(-) was the main DIC source used for photosynthesis. The M. chthonoplastes mats featured external and internal carbonic anhydrase (CA) activity as measured in intact cells and cell extracts, respectively. Acetazolamide (AZ, which slowly enters the cell and then inhibits mainly the external CA) and ethoxyzolamide (EZ, which inhibits both external and internal CA) reduced significantly the oxygen evolution rates, demonstrating that the CA was implied in the DIC acquisition. Vanadate inhibited photosynthesis by 60% although its application, when CA being inhibited (i.e. after applying AZ + EZ), did not produce any additional effect. It could indicate that ATPase-dependent HCO(3)(-) use occurred and also that this putative mechanism was coupled with CA like activity at the plasma membrane. The involvement of Na(+)-dependent HCO(3)( ) transporters in DIC acquisition was also inferred as monensin and 4-4' diisothiocyanatostibilene-2,2'-disulfonate (DIDS) reduced photosynthesis by 70%. DIDS produced a strong inhibitory effect even after application of AZ + EZ + vanadate, indicating that this mechanism was not related to CA activity. The microbial mats become subject to very unfavourable conditions for Rubisco carboxylation at their natural habitats (e.g. external pH of 10.5 and O(2) concentration doubled with respect to saturation concentration); therefore, this putative diversity of DIC acquisition mechanisms could ensure their growth under these extreme conditions. PMID- 18405334 TI - Expression and regulation of carbonic anhydrases in the marine diatom Thalassiosira pseudonana and in natural phytoplankton assemblages from Great Bay, New Jersey. AB - BLAST searches of expressed sequence tag libraries have revealed putative homologs of the archetypal diatom delta-carbonic anhydrase (CA) TWCA1 (for Thalassiosira weissflogii CA) in a broad range of eukaryotic phytoplankton including haptophytes, prasinophytes and dinoflagellates. Four putative homologs of TWCA1 are also reported and described from a search of the genomic sequence of Thalassiosira pseudonana and designated TWCA(Tp1-4). The delta-CA class is therefore more widely distributed in marine phytoplankton than previously thought. In particular, it is not restricted to the diatoms like the cadmium containing enzyme, CDCA, seems to be (zeta-CA class). Zinc status strongly influences growth rate in marine diatoms. We observed a decrease in the specific growth rate of T. pseudonana from 2.1 to 1.3 day(-1) when the unchelated Zn concentration (Zn') was lowered from 20 to 5 pM. In the same cultures, we detected a drop in whole-cell CA activity of about 50% in the most Zn-limited cells that occurred simultaneously with a large downregulation of TWCA(Tp1), TWCA(Tp2) and CDCA(Tp) gene transcripts and protein. These three genes were also found to be strongly upregulated by low pCO(2) in a manner typical of many algal CA enzymes. We have also conducted field experiments in diatom-dominated natural phytoplankton assemblages sampled from Great Bay of the coast of New Jersey. We observed increased total CA activity in parallel with increased expression of homologous twca and cdca gene transcripts in water incubated at increasingly higher pH values. Immunoblot and transcript expression analyses demonstrated a clear upregulation of CDCA transcript and protein as incubation pH increased both in the lab and in the field indicating that this CA is expressed under a broad range of environmental conditions and not restricted to low pCO(2) and low Zn. PMID- 18405335 TI - Inorganic carbon acquisition in potentially toxic and non-toxic diatoms: the effect of pH-induced changes in seawater carbonate chemistry. AB - The effects of pH-induced changes in seawater carbonate chemistry on inorganic carbon (C(i)) acquisition and domoic acid (DA) production were studied in two potentially toxic diatom species, Pseudo-nitzschia multiseries and Nitzschia navis-varingica, and the non-toxic Stellarima stellaris. In vivo activities of carbonic anhydrase (CA), photosynthetic O(2) evolution and CO(2) and HCO(3)(-) uptake rates were measured by membrane inlet MS in cells acclimated to low (7.9) and high pH (8.4 or 8.9). Species-specific differences in the mode of carbon acquisition were found. While extracellular carbonic anhydrase (eCA) activities increased with pH in P. multiseries and S. stellaris, N. navis-varingica exhibited low eCA activities independent of pH. Half-saturation concentrations (K(1/2)) for photosynthetic O(2) evolution, which were highest in S. stellaris and lowest in P. multiseries, generally decreased with increasing pH. In terms of carbon source, all species took up both CO(2) and HCO(3)(-). K(1/2) values for inorganic carbon uptake decreased with increasing pH in two species, while in N. navis-varingica apparent affinities did not change. While the contribution of HCO(3)(-) to net fixation was more than 85% in S. stellaris, it was about 55% in P. multiseries and only approximately 30% in N. navis-varingica. The intracellular content of DA increased in P. multiseries and N. navis-varingica with increasing pH. Based on our data, we propose a novel role for eCA acting as C(i)-recycling mechanism. With regard to pH-dependence of growth, the 'HCO(3)(-) user' S. stellaris was as sensitive as the 'CO(2) user' N. navis-varingica. The suggested relationship between DA and carbon acquisition/C(i) limitation could not be confirmed. PMID- 18405337 TI - A study on the use of male animal models for developing a live vaccine for brucellosis. AB - To study the safety of Brucella melitensis WR201, a live vaccine candidate, we compared the course of infection of this strain with that of virulent 16M in male BALB/c mice. At various times after oral immunization with strains WR201 or 16M, lungs, liver, spleen, testis, epididymis, inguinal and cervical lymph nodes were removed. Tissues were divided for microbiologic culture and histopathological examination. WR201 infection in male BALB/c mice had lower intensity and shorter duration than infection caused by virulent 16M. Pathological examination of testis and epididymis revealed no inflammation following strain WR201 immunization. In contrast, animals given virulent 16M strain had substantial inflammation in infected tissues. These data confirm the marked attenuation of WR201 relative to 16M. In addition, these studies suggest that male mice may be useful to assess the safety of live, attenuated Brucella vaccine candidates. PMID- 18405338 TI - Highly virulent porcine reproductive and respiratory syndrome virus emerged in China. AB - A highly pathogenic pig disease emerged in China in 2006, which was characterized by prolonged high fever, red discoloration of the body, and blue ears associated with high mortality. Porcine reproductive and respiratory syndrome virus (PRRSV) was isolated as the single most prominent virus in the samples collected from affected pigs. The full-length genomic sequence of the virus revealed two distinct deletions in the non-structural protein 2 (NSP2) in comparison to all previously reported North American genotype PRRSV. Through extensive surveys in 14 different provinces, 56 additional PRRSV isolates were obtained from affected farms. All of the isolates were found to contain identical deletions in NSP2. To confirm the etiology, eight 60-day-old PRRSV-free pigs were divided into two groups and the test group was intranasally infected at a titer of 2 x 10(5.0) tissue culture infectious dose 50 per pig. The inoculated pigs all died at 7, 8, 12, 16, or 21 days post-inoculation with their clinical and pathological findings similar to those in the field. The viruses recovered from dead pigs were identical to the inoculated virus in NSP2 and GP5 genes. Our study shows that the recently emerged PRRSV in China is characterized by two discontiguous deletions in NSP2 and is the cause for the current epizootics in China. PMID- 18405339 TI - Bacterial infections in pigs experimentally infected with Nipah virus. AB - Nipah virus (NiV; Paramyxoviridae) caused fatal encephalitis in humans during an outbreak in Malaysia in 1998/1999 after transmission from infected pigs. Our previous study demonstrated that the respiratory, lymphatic and central nervous systems are targets for virus replication in experimentally infected pigs. To continue the studies on pathogenesis of NiV in swine, six piglets were inoculated oronasally with 2.5 x 10(5) PFU per animal. Four pigs developed mild clinical signs, one exudative epidermitis, and one neurologic signs due to suppurative meningoencephalitis, and was euthanized at 11 days post-inoculation (dpi). Neutralizing antibodies reached in surviving animals titers around 1280 at 16 dpi. Nasal and oro-pharyngeal shedding of the NiV was detected between 2 and 17 dpi. Virus appeared to be cleared from the tissues of the infected animals by 23 dpi, with low amount of RNA detected in submandibular and bronchial lymph nodes of three pigs, and olfactory bulb of one animal. Despite the presence of neutralizing antibodies, virus was isolated from serum at 24 dpi, and the viral RNA was still detected in serum at 29 dpi. Our results indicate slower clearance of NiV from some of the infected pigs. Bacteria were detected in the cerebrospinal fluid of five NiV inoculated animals, with isolation of Streptococcus suis and Enterococcus faecalis. Staphylococcus hyicus was isolated from the skin lesions of the animal with exudative epidermitis. Along with the observed lymphoid depletion in the lymph nodes of all NiV-infected animals, and the demonstrated ability of NiV to infect porcine peripheral blood mononuclear cells in vitro, this finding warrants further investigation into a possible NiV induced immunosuppression of the swine host. PMID- 18405340 TI - Production, characterization and assay application of a purified, baculovirus expressed, serogroup specific bluetongue virus antigen. AB - The predominant serodiagnostic assay used in many countries to detect bluetongue virus (BTV) infections is a competitive enzyme-linked immunosorbent assay (c ELISA) which employs two critical reagents: a cell culture-derived BTV antigen and group-specific monoclonal antibody (Mab). Ongoing difficulties have been reported by laboratories in the production and quality control of the native antigen reagent which relies on the presence of adequate molar quantities and appropriate presentation of the major BTV core protein VP7. To address this important issue, a recombinant baculovirus was constructed containing a cDNA copy of genome segment 7 of BTV serotype 11 and used to infect insect cells which, in turn, expressed high levels of theVP7 protein with an estimated molecular mass of 39 kDa. In its purified form, this recombinant protein could be detected by group specific Mabs designated 3.17.A3 and 8A3B.6 produced against BTV serotypes 1 and 17, respectively, as well as by polyclonal bovine antibodies raised against North American and South African BTV serotypes. No reactivity was observed by Western blot analysis with these two Mabs suggesting that the common antigenic determinants, on the BTV VP7 protein, were mainly conformational. It was interesting to note that the purified recombinant VP7 protein demonstrated a greater degree of reactivity with Mab 8A3B.6 compared to that exhibited with Mab 3.17.A3 when evaluated in an ELISA. Due to its antigenic similarity to the native antigen, the recombinant protein was found to be a suitable replacement for use in a c-ELISA to detect BTV-specific antibodies with the added advantage that it could be consistently produced and was, therefore, amenable to quality control testing for purity, stability and other standards. PMID- 18405341 TI - Dermatophilus congolensis-associated placentitis, funisitis and abortion in a horse. AB - Placentitis, funisitis and fetal bronchopneumonia were diagnosed in an aborted full-term Thoroughbred fetus and its placenta by histopathological examination. Dermatophilus congolensis organisms were isolated from placenta, lung and stomach content. The genotypic identification of aerobic culture was confirmed by sequential analysis of the entire 16S rDNA gene. This is the first report of Dermatophilus congolensis-associated abortion in any species. PMID- 18405342 TI - Updates from the British Society for Investigative Dermatology Annual Meeting, 16 18 April 2007, Nottingham, U.K. AB - This is a synopsis of the main research findings presented at the British Society for Investigative Dermatology meeting held during 16-18 April 2007 in Nottingham, U.K. The conference highlighted the recent biological, epidemiological and therapeutic advances that have been made in the field of dermatology. The authors focus on the more important advances or summaries of findings, but this is not meant as a substitute for reading the conference proceedings and related references quoted in this article. PMID- 18405343 TI - A multifaceted role for polyamines in bacterial pathogens. AB - Polyamines are polycationic molecules with a hydrocarbon backbone and multiple amino groups. Descriptions of the physiological roles of polyamines have often been limited to their interaction with negatively charged nucleic acids. Of late, reports linking polyamines to microbial carcinogenesis, biofilm formation, escape from phagolysosomes, bacteriocin production, toxin activity and protection from oxidative and acid stress have been published, providing insights about their other important but lesser known functions. This review focuses on recently discovered novel functions of polyamines in microorganisms, with an emphasis on bacterial pathogens of humans. PMID- 18405345 TI - Protocol for SAMS (Support and Advice for Medication Study): a randomised controlled trial of an intervention to support patients with type 2 diabetes with adherence to medication. AB - BACKGROUND: Although some interventions have been shown to improve adherence to medication for diabetes, results are not consistent. We have developed a theory based intervention which we will evaluate in a well characterised population to test efficacy and guide future intervention development and trial design. METHODS AND DESIGN: The SAMS (Supported Adherence to Medication Study) trial is a primary care based multi-centre randomised controlled trial among 200 patients with type 2 diabetes and an HbA1c of 7.5% or above. It is designed to evaluate the efficacy of a two-component motivational intervention based on the Theory of Planned Behaviour and volitional action planning to support medication adherence compared with standard care. The intervention is delivered by practice nurses. Nurses were trained using a workshop approach with role play and supervised using assessment of tape-recorded consultations. The trial has a two parallel groups design with an unbalanced three-to-two individual randomisation eight weeks after recruitment with twelve week follow-up. The primary outcome is medication adherence measured using an electronic medication monitor over 12 weeks and expressed as the difference between intervention and control in mean percentage of days on which the correct number of medication doses is taken. Subgroup analyses will explore impact of number of medications taken, age, HbA1c, and self-reported adherence at baseline on outcomes. The study also measures the effect of dispensing medication to trial participants packaged in the electronic medication-monitoring device compared with conventional medication packaging. This will be achieved through one-to-one randomisation at recruitment to these conditions with assessment of the difference between groups in self-report of medication adherence and change in mean HbA1c from baseline to eight weeks. Anonymised demographic data are collected on non-respondents. Central randomisation is carried out independently of trial co-ordination and practices using minimisation to adjust for selected confounders. DISCUSSION: The SAMS intervention and trial design address weaknesses of previous research by recruitment from a well-characterised population, definition of a feasible theory based intervention to support medication taking and careful measurement to estimate and interpret efficacy. The results will inform practice and the design of a cost-effectiveness trial [ISRCTN30522359]. PMID- 18405344 TI - Transcription factors GATA-4 and GATA-6 in normal and neoplastic human gastrointestinal mucosa. AB - BACKGROUND: Human gastrointestinal mucosa regenerates vigorously throughout life, but the factors controlling cell fate in mature mucosa are poorly understood. GATA transcription factors direct cell proliferation and differentiation in many organs, and are implicated in tumorigenesis. GATA-4 and GATA-6 are considered crucial for the formation of murine gastrointestinal mucosa, but their role in human gastrointestinal tract remains unexplored. We studied in detail the expression patterns of these two GATA factors and a GATA-6 down-stream target, Indian hedgehog (Ihh), in normal human gastrointestinal mucosa. Since these factors are considered important for proliferation and differentiation, we also explored the possible alterations in their expression in gastrointestinal neoplasias. The expression of the carcinogenesis-related protein Indian hedgehog was also investigated in comparison to GATA factors. METHODS: Samples of normal and neoplastic gastrointestinal tract from children and adults were subjected to RNA in situ hybridization with 33P labelled probes and immunohistochemistry, using an avidin-biotin immunoperoxidase system. The pathological tissues examined included samples of chronic and atrophic gastritis as well as adenomas and adenocarcinomas of the colon and rectum. RESULTS: GATA-4 was abundant in the differentiated epithelial cells of the proximal parts of the gastrointestinal tract but was absent from the distal parts. In contrast, GATA-6 was expressed throughout the gastrointestinal epithelium, and in the distal gut its expression was most intense at the bottom of the crypts, i.e. cells with proliferative capacity. Both factors were also present in Barrett's esophagus and metaplasia of the stomach. GATA-6 expression was reduced in colon carcinoma. Ihh expression overlapped with that of GATA-6 especially in benign gastrointestinal neoplasias. CONCLUSION: The results suggest differential but overlapping functions for GATA-4 and GATA-6 in the normal gastrointestinal mucosa. Furthermore, GATA-4, GATA-6 and Ihh expression is altered in premalignant dysplastic lesions and reduced in overt cancer. PMID- 18405346 TI - Analysis of Aspergillus nidulans metabolism at the genome-scale. AB - BACKGROUND: Aspergillus nidulans is a member of a diverse group of filamentous fungi, sharing many of the properties of its close relatives with significance in the fields of medicine, agriculture and industry. Furthermore, A. nidulans has been a classical model organism for studies of development biology and gene regulation, and thus it has become one of the best-characterized filamentous fungi. It was the first Aspergillus species to have its genome sequenced, and automated gene prediction tools predicted 9,451 open reading frames (ORFs) in the genome, of which less than 10% were assigned a function. RESULTS: In this work, we have manually assigned functions to 472 orphan genes in the metabolism of A. nidulans, by using a pathway-driven approach and by employing comparative genomics tools based on sequence similarity. The central metabolism of A. nidulans, as well as biosynthetic pathways of relevant secondary metabolites, was reconstructed based on detailed metabolic reconstructions available for A. niger and Saccharomyces cerevisiae, and information on the genetics, biochemistry and physiology of A. nidulans. Thereby, it was possible to identify metabolic functions without a gene associated, and to look for candidate ORFs in the genome of A. nidulans by comparing its sequence to sequences of well-characterized genes in other species encoding the function of interest. A classification system, based on defined criteria, was developed for evaluating and selecting the ORFs among the candidates, in an objective and systematic manner. The functional assignments served as a basis to develop a mathematical model, linking 666 genes (both previously and newly annotated) to metabolic roles. The model was used to simulate metabolic behavior and additionally to integrate, analyze and interpret large-scale gene expression data concerning a study on glucose repression, thereby providing a means of upgrading the information content of experimental data and getting further insight into this phenomenon in A. nidulans. CONCLUSION: We demonstrate how pathway modeling of A. nidulans can be used as an approach to improve the functional annotation of the genome of this organism. Furthermore we show how the metabolic model establishes functional links between genes, enabling the upgrade of the information content of transcriptome data. PMID- 18405347 TI - Electrospun micro- and nanofiber tubes for functional nervous regeneration in sciatic nerve transections. AB - BACKGROUND: Although many nerve prostheses have been proposed in recent years, in the case of consistent loss of nervous tissue peripheral nerve injury is still a traumatic pathology that may impair patient's movements by interrupting his motor sensory pathways. In the last few decades tissue engineering has opened the door to new approaches;: however most of them make use of rigid channel guides that may cause cell loss due to the lack of physiological local stresses exerted over the nervous tissue during patient's movement. Electrospinning technique makes it possible to spin microfiber and nanofiber flexible tubular scaffolds composed of a number of natural and synthetic components, showing high porosity and remarkable surface/volume ratio. RESULTS: In this study we used electrospun tubes made of biodegradable polymers (a blend of PLGA/PCL) to regenerate a 10-mm nerve gap in a rat sciatic nerve in vivo. Experimental groups comprise lesioned animals (control group) and lesioned animals subjected to guide conduits implantated at the severed nerve stumps, where the tubular scaffolds are filled with saline solution. Four months after surgery, sciatic nerves failed to reconnect the two stumps of transected nerves in the control animal group. In most of the treated animals the electrospun tubes induced nervous regeneration and functional reconnection of the two severed sciatic nerve tracts. Myelination and collagen IV deposition have been detected in concurrence with regenerated fibers. No significant inflammatory response has been found. Neural tracers revealed the re establishment of functional neuronal connections and evoked potential results showed the reinnervation of the target muscles in the majority of the treated animals. CONCLUSION: Corroborating previous works, this study indicates that electrospun tubes, with no additional biological coating or drug loading treatment, are promising scaffolds for functional nervous regeneration. They can be knitted in meshes and various frames depending on the cytoarchitecture of the tissue to be regenerated. The versatility of this technique gives room for further scaffold improvements, like tuning the mechanical properties of the tubular structure or providing biomimetic functionalization. Moreover, these guidance conduits can be loaded with various fillers like collagen, fibrin, or self-assembling peptide gels or loaded with neurotrophic factors and seeded with cells. Electrospun scaffolds can also be synthesized in different micro architectures to regenerate lesions in other tissues like skin and bone. PMID- 18405348 TI - Prevalence of chronic kidney disease in population-based studies: systematic review. AB - BACKGROUND: Chronic kidney disease (CKD) is becoming a major public health problem worldwide. This article reviews the published evidence of prevalence of CKD in population-based study samples that used the standardized definition from the Kidney Disease Outcomes Quality Initiative of the National Kidney Foundation (K/DOQI) practice guideline, and particularly focus on performance of serum creatinine based equations for GFR estimation. We provide a summary of available data about the burden of CKD in various populations. METHODS: We performed a systematic review of available published data in MEDLINE. A combination of various keywords relevant to CKD was used in this research. Related data of included studies were extracted in a systematic way. RESULTS: A total of 26 studies were included in this review. The studies were conducted in different populations, and the number of study participants ranged from 237 to 65181. The median prevalence of CKD was 7.2% in persons aged 30 years or older. In persons aged 64 years or older prevalence of CKD varied from 23.4% to 35.8%. Importantly, the prevalence of CKD strongly depended on which estimating equations were used. The Modification of Diet in Renal Disease Study (MDRD) equation was likely to be preferred in recent epidemiological studies compared to the adjusted Cockcroft Gault (CG) equation. CONCLUSION: Worldwide, CKD is becoming a common disease in the general population. Accurately detecting CKD in special groups remains inadequate, particularly among elderly persons, females or other ethnic groups such as Asians. PMID- 18405349 TI - Array-CGH in patients with Kabuki-like phenotype: identification of two patients with complex rearrangements including 2q37 deletions and no other recurrent aberration. AB - BACKGROUND: Kabuki syndrome (KS) is a multiple congenital anomaly syndrome characterized by specific facial features, mild to moderate mental retardation, postnatal growth delay, skeletal abnormalities, and unusual dermatoglyphic patterns with prominent fingertip pads. A 3.5 Mb duplication at 8p23.1-p22 was once reported as a specific alteration in KS but has not been confirmed in other patients. The molecular basis of KS remains unknown. METHODS: We have studied 16 Spanish patients with a clinical diagnosis of KS or KS-like to search for genomic imbalances using genome-wide array technologies. All putative rearrangements were confirmed by FISH, microsatellite markers and/or MLPA assays, which also determined whether the imbalance was de novo or inherited. RESULTS: No duplication at 8p23.1-p22 was observed in our patients. We detected complex rearrangements involving 2q in two patients with Kabuki-like features: 1) a de novo inverted duplication of 11 Mb with a 4.5 Mb terminal deletion, and 2) a de novo 7.2 Mb-terminal deletion in a patient with an additional de novo 0.5 Mb interstitial deletion in 16p. Additional copy number variations (CNV), either inherited or reported in normal controls, were identified and interpreted as polymorphic variants. No specific CNV was significantly increased in the KS group. CONCLUSION: Our results further confirmed that genomic duplications of 8p23 region are not a common cause of KS and failed to detect other recurrent rearrangement causing this disorder. The detection of two patients with 2q37 deletions suggests that there is a phenotypic overlap between the two conditions, and screening this region in the Kabuki-like patients should be considered. PMID- 18405350 TI - Prognostic impact of array-based genomic profiles in esophageal squamous cell cancer. AB - BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a genetically complex tumor type and a major cause of cancer related mortality. Although distinct genetic alterations have been linked to ESCC development and prognosis, the genetic alterations have not gained clinical applicability. We applied array based comparative genomic hybridization (aCGH) to obtain a whole genome copy number profile relevant for identifying deranged pathways and clinically applicable markers. METHODS: A 32 k aCGH platform was used for high resolution mapping of copy number changes in 30 stage I-IV ESCC. Potential interdependent alterations and deranged pathways were identified and copy number changes were correlated to stage, differentiation and survival. RESULTS: Copy number alterations affected median 19% of the genome and included recurrent gains of chromosome regions 5p, 7p, 7q, 8q, 10q, 11q, 12p, 14q, 16p, 17p, 19p, 19q, and 20q and losses of 3p, 5q, 8p, 9p and 11q. High-level amplifications were observed in 30 regions and recurrently involved 7p11 (EGFR), 11q13 (MYEOV, CCND1, FGF4, FGF3, PPFIA, FAD, TMEM16A, CTTS and SHANK2) and 11q22 (PDFG). Gain of 7p22.3 predicted nodal metastases and gains of 1p36.32 and 19p13.3 independently predicted poor survival in multivariate analysis. CONCLUSION: aCGH profiling verified genetic complexity in ESCC and herein identified imbalances of multiple central tumorigenic pathways. Distinct gains correlate with clinicopathological variables and independently predict survival, suggesting clinical applicability of genomic profiling in ESCC. PMID- 18405351 TI - Doctor's perception of doctor-patient relationships in emergency departments: What roles do gender and ethnicity play? AB - BACKGROUND: Emergency departments continuously provide medical treatment on a walk-in basis. Several studies investigated the patient's perception of the doctor-patient relationship, but few have asked doctors about their views. Furthermore, the influence of the patient's ethnicity and gender on the doctor's perception remains largely unanswered. METHODS: Based on data collated in three gynaecology (GYN)/internal medicine (INT) emergency departments in Berlin, Germany, we evaluated the impact of the patient's gender and ethnicity on the doctors' satisfaction with the course of the treatment they provided. Information was gathered from 2.429 short questionnaires completed by doctors and the medical records of the corresponding patients. RESULTS: The patient's ethnicity had a significant impact on the doctors' satisfaction with the doctor-patient relationship. Logistic regression analysis showed that the odds ratio (OR) for physician satisfaction was significantly lower for patients of Turkish origin (OR = 2.6 INT and 5.5 GYN) than for those of German origin. The main reasons stated were problems with communication and a perceived lack of urgency for emergency treatment. The odds ratios for dissatisfaction due to a lack of language skills were 4.48 (INT) and 6.22 (GYN), and those due to perceived lack of urgency for emergency treatment were 0.75 (INT) and 0.63 (GYN). Sex differences caused minor variation. CONCLUSION: The results show that good communication despite language barriers is crucial in providing medical care that is satisfactory to both patient and doctors, especially in emergency situations. Therefore the use of professional interpreters for improved communication and the training of medical staff for improved intercultural competence are essential for the provision of adequate health care in a multicultural setting. PMID- 18405352 TI - Animal-assisted therapy with farm animals for persons with psychiatric disorders: effects on self-efficacy, coping ability and quality of life, a randomized controlled trial. AB - BACKGROUND: The benefits of Animal-Assisted Therapy (AAT) for humans with mental disorders have been well-documented using cats and dogs, but there is a complete lack of controlled studies using farm animals as therapeutic agents for psychiatric patients. The study was developed in the context of Green care, a concept that involves the use of farm animals, plants, gardens, or the landscape in recreational or work-related interventions for different target groups of clients in cooperation with health authorities. The present study aimed at examining effects of a 12-week intervention with farm animals on self-efficacy, coping ability and quality of life among adult psychiatric patients with a variety of psychiatric diagnoses. METHODS: The study was a randomized controlled trial and follow-up. Ninety patients (59 women and 31 men) with schizophrenia, affective disorders, anxiety, and personality disorders completed questionnaires to assess self-efficacy (Generalized Self-Efficacy Scale; GSE), coping ability (Coping Strategies Scale), and quality of life (Quality of Life Scale; QOLS-N) before, at the end of intervention, and at six months follow-up. Two-thirds of the patients (N = 60) were given interventions; the remaining served as controls. RESULTS: There was significant increase in self-efficacy in the treatment group but not in the control group from before intervention (SB) to six months follow up (SSMA), (SSMA-SB; F1,55 = 4.20, p= 0.05) and from end of intervention (SA) to follow-up (SSMA-SA; F1,55 = 5.6, p= 0.02). There was significant increase in coping ability within the treatment group between before intervention and follow up (SSMA-SB = 2.7, t = 2.31, p = 0.03), whereas no changes in quality of life was found. There were no significant changes in any of the variables during the intervention. CONCLUSION: AAT with farm animals may have positive influences on self-efficacy and coping ability among psychiatric patients with long lasting psychiatric symptoms. PMID- 18405353 TI - Unit-specific calibration of Actigraph accelerometers in a mechanical setup - is it worth the effort? The effect on random output variation caused by technical inter-instrument variability in the laboratory and in the field. AB - BACKGROUND: Potentially, unit-specific in-vitro calibration of accelerometers could increase field data quality and study power. However, reduced inter-unit variability would only be important if random instrument variability contributes considerably to the total variation in field data. Therefore, the primary aim of this study was to calculate and apply unit-specific calibration factors in multiple accelerometers in order to examine the impact on random output variation caused by inter-instrument variability. METHODS: Instrument-specific calibration factors were estimated in 25 MTI- and 53 CSA accelerometers in a mechanical setup using four different settings varying in frequencies and/or amplitudes. Calibration effect was analysed by comparing raw and calibrated data after applying unit-specific calibration factors to data obtained during quality checks in a mechanical setup and to data collected during free living conditions. RESULTS: Calibration reduced inter-instrument variability considerably in the mechanical setup, both in the MTI instruments (raw SDbetween units = 195 counts*min-1 vs. calibrated SDbetween units = 65 counts*min-1) and in the CSA instruments (raw SDbetween units = 343 counts*min-1 vs. calibrated SDbetween units = 67 counts*min-1). However, the effect of applying the derived calibration to children's and adolescents' free living physical activity data did not alter the coefficient of variation (CV) (children: CVraw = 30.2% vs. CVcalibrated = 30.4%, adolescents: CVraw = 36.3% vs. CVcalibrated = 35.7%). High correlations (r = 0.99 & r = 0.98, respectively) were observed between raw and calibrated field data, and the proportion of the total variation caused by the MTI- and CSA monitor was estimated to be only 1.1% and 4.2%, respectively. Compared to the CSA instruments, a significantly increased (9.95%) mean acceleration response was observed post hoc in the batch of MTI instruments, in which a significantly reduced inter-instrumental reliability was observed over time. CONCLUSION: The application of unit-specific calibration factors to data collected during free living conditions had no apparent effect on inter-instrument variability. In all probability, the effect of technical calibration was primarily attenuated in the field by other more dominant sources of variation. However, routine technical assessments are still very important for determining the acceleration responses in the batch of instruments being used and, if performed after every field use, for preventing decidedly broken instruments from being returned into the field repeatedly. PMID- 18405354 TI - Natural course of intra-articular shifting bone marrow edema syndrome of the knee. AB - BACKGROUND: Intra-articular shift (migration) of bone marrow edema syndrome (BMES) is a very rare disease. Only a few cases have been reported thus far. The condition may cause the clinician to suspect an aggressive disease. METHODS: We reviewed eight patients (four women and four men) with unilateral BMES located in the knee. The patients were aged 39 to 56 years (mean, 49.2 years). In all patients, bone marrow edema (BME) initially observed on magnetic resonance imaging (MR imaging) shifted within the same joint, i.e. from the medial to the lateral femoral condyle or the adjacent bone. Seven patients were given conservative therapy, including limited weight-bearing, for a period of three weeks after the initial detection of BMES, whereas one patient underwent surgical core decompression twice. RESULTS: MR imaging showed complete restitution in 6 cases and a small residual edema in one case. A final control MR could not be obtained for one patient, who had no pain. A further patient had an avascular necrosis of the contralateral hip after 16 months. Improvement on MR imaging was correlated with the clinical outcome in all cases. All patients became asymptomatic after a mean period of 9 months (6-11). INTERPRETATION: Intra articular shifting BMES is a very rare condition. As the disease is self limiting, conservative therapy may be recommended. PMID- 18405355 TI - Complex evolution of the DAL5 transporter family. AB - BACKGROUND: Genes continuously duplicate and the duplicated copies remain in the genome or get deleted. The DAL5 subfamily of transmembrane transporter genes has eight known members in S. cerevisiae. All are putative anion:cation symporters of vitamins (such as allantoate, nicotinate, panthotenate and biotin). The DAL5 subfamily is an old and important group since it is represented in both Basidiomycetes ("mushrooms") and Ascomycetes ("yeast"). We studied the complex evolution of this group in species from the kingdom of fungi particularly the Ascomycetes. RESULTS: We identified numerous gene duplications creating sets of orthologous and paralogous genes. In different lineages the DAL5 subfamily members expanded or contracted and in some lineages a specific member could not be found at all. We also observed a close relationship between the gene YIL166C and its homologs in the Saccharomyces sensu stricto species and two "wine spoiler" yeasts, Dekkera bruxellensis and Candida guilliermondi, which could possibly be the result of horizontal gene transfer between these distantly related species. In the analyses we detect several well defined groups without S. cerevisiae representation suggesting new gene members in this subfamily with perhaps altered specialization or function. CONCLUSION: The transmembrane DAL5 subfamily was found to have a very complex evolution in yeast with intra- and interspecific duplications and unusual relationships indicating specialization, specific deletions and maybe even horizontal gene transfer. We believe that this group will be important in future investigations of evolution in fungi and especially the evolution of transmembrane proteins and their specialization. PMID- 18405356 TI - The human ABC transporter pseudogene family: Evidence for transcription and gene pseudogene interference. AB - BACKGROUND: Pseudogenes are an integral component of the human genome. Little attention, however, has so far been paid to the phenomenon that some pseudogenes are transcriptionally active. Recently, we demonstrated that the human ortholog of the rodent testis-specific ATP-binding cassette (ABC) transporter Abca17 is a ubiquitously transcribed pseudogene (ABCA17P). The aim of the present study was to establish a complete inventory of all ABC transporter pseudogenes in the human genome and to identify transcriptionally active ABC transporter pseudogenes. Moreover, we tested the hypothesis that a regulatory interdependency exists between ABC transporter pseudogenes and their parental protein coding equivalents. RESULTS: Systematic bioinformatic analysis revealed the existence of 22 ABC transporter pseudogenes within the human genome. We identified two clusters on chromosomes 15 and 16, respectively, which harbor almost half of all pseudogenes (n = 10). Available information from EST and mRNA databases and RT PCR expression profiling indicate that a large portion of the ABC transporter pseudogenes (45%, n = 10) are transcriptionally active and some of them are expressed as alternative splice variants. We demonstrate that both pseudogenes of the pseudoxanthoma elasticum gene ABCC6, ABCC6P1 and ABCC6P2, are transcribed. ABCC6P1 and ABCC6 possess near-identical promoter sequences and their tissue specific expression profiles are strikingly similar raising the possibility that they form a gene-pseudogene dual transcription unit. Intriguingly, targeted knockdown of the transcribed pseudogene ABCC6P1 resulted in a significant reduction of ABCC6 mRNA expression levels. CONCLUSION: The human genome contains a surprisingly small number of ABC transporter pseudogenes relative to other known gene families. They are unevenly distributed across the chromosomes. Importantly, a significant portion of the ABC transporter pseudogenes is transcriptionally active. The downregulation of ABCC6 mRNA levels by targeted suppression of the expression of its pseudogene ABCC6P1 provides evidence, for the first time, for a regulatory interdependence of a transcribed pseudogene and its protein coding counterpart in the human genome. PMID- 18405357 TI - Graphical presentation of diagnostic information. AB - BACKGROUND: Graphical displays of results allow researchers to summarise and communicate the key findings of their study. Diagnostic information should be presented in an easily interpretable way, which conveys both test characteristics (diagnostic accuracy) and the potential for use in clinical practice (predictive value). METHODS: We discuss the types of graphical display commonly encountered in primary diagnostic accuracy studies and systematic reviews of such studies, and systematically review the use of graphical displays in recent diagnostic primary studies and systematic reviews. RESULTS: We identified 57 primary studies and 49 systematic reviews. Fifty-six percent of primary studies and 53% of systematic reviews used graphical displays to present results. Dot-plot or box and- whisker plots were the most commonly used graph in primary studies and were included in 22 (39%) studies. ROC plots were the most common type of plot included in systematic reviews and were included in 22 (45%) reviews. One primary study and five systematic reviews included a probability-modifying plot. CONCLUSION: Graphical displays are currently underused in primary diagnostic accuracy studies and systematic reviews of such studies. Diagnostic accuracy studies need to include multiple types of graphic in order to provide both a detailed overview of the results (diagnostic accuracy) and to communicate information that can be used to inform clinical practice (predictive value). Work is required to improve graphical displays, to better communicate the utility of a test in clinical practice and the implications of test results for individual patients. PMID- 18405358 TI - Autosomal mutations affecting Y chromosome loops in Drosophila melanogaster. AB - BACKGROUND: The Y chromosome of Drosophila melanogaster harbors several genes required for male fertility. The genes for these fertility factors are very large in size and contain conspicuous amounts of repetitive DNA and transposons. Three of these loci (ks-1, kl-3 and kl-5) have the ability to develop giant lampbrush like loops in primary spermatocytes, a cytological manifestation of their active state in these cells. Y-loops bind a number of non-Y encoded proteins, but the mechanisms regulating their development and their specific functions are still to be elucidated. RESULTS: Here we report the results of a screen of 726 male sterile lines to identify novel autosomal genes controlling Y-loop function. We analyzed mutant testis preparations both in vivo and by immunofluorescence using antibodies directed against Y-loop-associated proteins. This screen enabled us to isolate 17 mutations at 15 loci whose wild-type function is required for proper Y loop morphogenesis. Six of these loci are likely to specifically control loop development, while the others display pleiotropic effects on both loops and meiotic processes such as spermiogenesis, sperm development and maturation. We also determined the map position of the mutations affecting exclusively Y-loop morphology. CONCLUSION: Our cytological screening permitted us to identify novel genetic functions required for male spermatogenesis, some of which show pleiotropic effects. Analysis of these mutations also shows that loop development can be uncoupled from meiosis progression. These data represent a useful framework for the characterization of Y-loop development at a molecular level and for the study of the genetic control of heterochromatin. PMID- 18405359 TI - The Dutch version of the Oral Health Impact Profile (OHIP-NL): Translation, reliability and construct validity. AB - BACKGROUND: The purpose of this study was to make a cross-culturally adapted, Dutch version of the Oral Health Impact Profile (OHIP), a 49-item questionnaire measuring oral health-related quality of life, and to examine its psychometric properties. METHODS: The original English version of the OHIP was translated into the Dutch language, following the guidelines for cross-cultural adaptation of health-related quality of life measures. The resulting OHIP-NL's psychometric properties were examined in a sample of 119 patients (68.9 % women; mean age = 57.1 +/- 12.2 yrs). They were referred to the clinic of Prosthodontics and Implantology with complaints concerning their partial or full dentures or other problems with missing teeth. To establish the reliability of the OHIP-NL, internal consistency and test-retest reliability (N = 41; 1 - 2 weeks interval) were examined, using Cronbach's alpha and intraclass correlation coefficients (ICC), respectively. Further, construct validity was established by calculating ANOVA. RESULTS: Internal consistency and test-retest reliability were excellent (Cronbach's alpha = 0.82 - 0.97; ICC = 0.78 - 0.90). In addition, all associations were significant and in the expected direction. CONCLUSION: IN CONCLUSION: the OHIP-NL can be considered a reliable and valid instrument to measure oral health-related quality of life. PMID- 18405360 TI - Interpreting changes in measles genotype: the contribution of chance, migration and vaccine coverage. AB - BACKGROUND: In some populations, complete shifts in the genotype of the strain of measles circulating in the population have been observed, with given genotypes being replaced by new genotypes. Studies have postulated that such shifts may be attributable to differences between the fitness of the new and the old genotypes. METHODS: We developed a stochastic model of the transmission dynamics of measles, simulating the effects of different levels of migration, vaccination coverage and importation of new genotypes on patterns in the persistence and replacement of indigenous genotypes. RESULTS: The analyses illustrate that complete replacement in the genotype of the strain circulating in populations may occur because of chance. This occurred in >50% of model simulations, for levels of vaccination coverage and numbers of imported cases per year which are compatible with those observed in several Western European populations (>80% and >3 per million per year respectively) and for the given assumptions in the model. CONCLUSION: The interpretation of genotypic data, which are increasingly being collected in surveillance programmes, needs to take account of the underlying vaccination coverage and the level of the importation rate of measles cases into the population. PMID- 18405361 TI - Using an evolutionary algorithm and parallel computing for haplotyping in a general complex pedigree with multiple marker loci. AB - BACKGROUND: Haplotype reconstruction is important in linkage mapping and association mapping of quantitative trait loci (QTL). One widely used statistical approach for haplotype reconstruction is simulated annealing (SA), implemented in SimWalk2. However, the algorithm needs a very large number of sequential iterations, and it does not clearly show if convergence of the likelihood is obtained. RESULTS: An evolutionary algorithm (EA) is a good alternative whose convergence can be easily assessed during the process. It is feasible to use a powerful parallel-computing strategy with the EA, increasing the computational efficiency. It is shown that the EA can be approximately 4 times faster and gives more reliable estimates than SimWalk2 when using 4 processors. In addition, jointly updating dependent variables can increase the computational efficiency up to approximately 2 times. Overall, the proposed method with 4 processors increases the computational efficiency up to approximately 8 times compared to SimWalk2. The efficiency will increase more with a larger number of processors. CONCLUSION: The use of the evolutionary algorithm and the joint updating method can be a promising tool for haplotype reconstruction in linkage and association mapping of QTL. PMID- 18405362 TI - Occurrence of cancer at multiple sites: towards distinguishing multigenesis from metastasis. AB - BACKGROUND: Occurrence of tumors at multiple sites is a hallmark of malignant cancers and contributes to the high mortality of cancers. The formation of multi site cancers (MSCs) has conventionally been regarded as a result of hematogenous metastasis. However, some MSCs may appear as unusual in the sense of vascular dissemination pattern and therefore be explained by alternative metastasis models or even by non-metastatic independent formation mechanisms. RESULTS: Through literature review and incorporation of recent advance in understanding aging and development, we identified two alternative mechanisms for the independent formation of MSCs: 1) formation of separate tumors from cancer-initiating cells (CICs) mutated at an early stage of development and then diverging as to their physical locations upon further development, 2) formation of separate tumors from different CICs that contain mutations in some convergent ways. Either of these processes does not require long-distance migration and/or vascular dissemination of cancer cells from a primary site to a secondary site. Thus, we classify the formation of these MSCs from indigenous CICs (iCICs) into a new mechanistic category of tumor formation - multigenesis. CONCLUSION: A multigenesis view on multi-site cancer (MSCs) may offer explanations for some "unusual metastasis" and has important implications for designing expanded strategies for the diagnosis and treatment of cancers. PMID- 18405363 TI - Coexpression of GM-CSF and antigen in DNA prime-adenoviral vector boost immunization enhances polyfunctional CD8+ T cell responses, whereas expression of GM-CSF antigen fusion protein induces autoimmunity. AB - BACKGROUND: Granulocyte-macrophage colony-stimulating factor (GM-CSF) has shown promising results as a cytokine adjuvant for antiviral vaccines and in various models of tumor gene therapy. To explore whether the targeting of antigens to GM CSF receptors on antigen-presenting cells enhances antigen-specific CD8 T-cell responses, fusion proteins of GM-CSF and ovalbumin (OVA) were expressed by DNA and adenoviral vector vaccines. In addition, bicistronic vectors allowing independent expression of the antigen and the cytokine were tested in parallel. RESULTS: In vitro, the GM-CSF ovalbumin fusion protein (GM-OVA) led to the better stimulation of OVA-specific CD8+ T cells by antigen-presenting cells than OVA and GM-CSF given as two separate proteins. However, prime-boost immunizations of mice with DNA and adenoviral vector vaccines encoding GM-OVA suppressed CD8+ T-cell responses to OVA. OVA-specific IgG2a antibody levels were also reduced, while the IgG1 antibody response was enhanced. Suppression of CD8+ T cell responses by GM OVA vaccines was associated with the induction of neutralizing antibodies to GM CSF. In contrast, the coexpression of GM-CSF and antigens in DNA prime adenoviral boost immunizations led to a striking expansion of polyfunctional OVA-specific CD8+ T cells without the induction of autoantibodies. CONCLUSION: The induction of autoantibodies suggests a general note of caution regarding the use of highly immunogenic viral vector vaccines encoding fusion proteins between antigens and host proteins. In contrast, the expansion of polyfunctional OVA-specific CD8+ T cells after immunizations with bicistronic vectors further support a potential application of GM-CSF as an adjuvant for heterologous prime-boost regimens with genetic vaccines. Since DNA prime adenoviral vector boost regimenes are presently considered as one of the most efficient ways to induce CD8+ T cell responses in mice, non-human primates and humans, further enhancement of this response by GM CSF is a striking observation. PMID- 18405364 TI - A unifying hypothesis for hydrocephalus, Chiari malformation, syringomyelia, anencephaly and spina bifida. AB - This work is a modified version of the Casey Holter Memorial prize essay presented to the Society for Research into Hydrocephalus and Spina Bifida, June 29th 2007, Heidelberg, Germany. It describes the origin and consequences of the Chiari malformation, and proposes that hydrocephalus is caused by inadequate central nervous system (CNS) venous drainage. A new hypothesis regarding the pathogenesis, anencephaly and spina bifida is described.Any volume increase in the central nervous system can increase venous pressure. This occurs because veins are compressible and a CNS volume increase may result in reduced venous blood flow. This has the potential to cause progressive increase in cerebrospinal fluid (CSF) volume. Venous insufficiency may be caused by any disease that reduces space for venous volume. The flow of CSF has a beneficial effect on venous drainage. In health it moderates central nervous system pressure by moving between the head and spine. Conversely, obstruction to CSF flow causes localised pressure increases, which have an adverse effect on venous drainage.The Chiari malformation is associated with hindbrain herniation, which may be caused by low spinal pressure relative to cranial pressure. In these instances, there are hindbrain-related symptoms caused by cerebellar and brainstem compression. When spinal injury occurs as a result of a Chiari malformation, the primary pathology is posterior fossa hypoplasia, resulting in raised spinal pressure. The small posterior fossa prevents the flow of CSF from the spine to the head as blood enters the central nervous system during movement. Consequently, intermittent increases in spinal pressure caused by movement, result in injury to the spinal cord. It is proposed that posterior fossa hypoplasia, which has origins in fetal life, causes syringomyelia after birth and leads to damage to the spinal cord in spina bifida. It is proposed that hydrocephalus may occur as a result of posterior fossa hypoplasia, where raised pressure occurs as a result of obstruction to flow of CSF from the head to the spine, and cerebral injury with raised pressure occurs in anencephaly by this mechanism.The current view of dysraphism is that low central nervous system pressure and exposure to amniotic fluid, damage the central nervous system. The hypothesis proposed in this essay supports the view that spina bifida is a manifestation of progressive hydrocephalus in the fetus. It is proposed that that mesodermal growth insufficiency influences both neural tube closure and central nervous system pressure, leading to dysraphism. PMID- 18405365 TI - Intra-articular temperatures of the knee in sports - an in-vivo study of jogging and alpine skiing. AB - BACKGROUND: Up to date, no information exists about the intra-articular temperature changes of the knee related to activity and ambient temperature. METHODS: In 6 healthy males, a probe for intra-articular measurement was inserted into the notch of the right knee. Each subject was jogging on a treadmill in a closed room at 19 degrees C room temperature and skiing in a ski resort at -3 degrees C outside temperature for 60 minutes. In both conditions, temperatures were measured every fifteen minutes intra-articulary and at the skin surface of the knee. A possible influence on joint function and laxity was evaluated before and after activity. Statistical analysis of intra-articular and skin temperatures was done using nonparametric Wilcoxon's sign rank sum test and Mann-Whitney's-U Test. RESULTS: Median intra-articular temperatures increased from 31.4 degrees C before activity by 2.1 degrees C, 4 degrees C, 5.8 degrees C and 6.1 degrees C after 15, 30, 45 and 60 min of jogging (all p < or = 0.05). Median intra articular temperatures dropped from 32.2 degrees C before activity by 0.5 degrees C, 1.9 degrees C, 3.6 degrees C and 1.1 degrees C after 15, 30, 45 and 60 min of skiing (all n.s.). After 60 minutes of skiing (jogging), the median intra articular temperature was 19.6% (8.7%) higher than the skin surface temperature at the knee. Joint function and laxity appeared not to be different before and after activity within both groups. CONCLUSION: This study demonstrates different changes of intra-articular and skin temperatures during sports in jogging and alpine skiing and suggests that changes are related to activity and ambient temperature. PMID- 18405366 TI - Local conservation scores without a priori assumptions on neutral substitution rates. AB - BACKGROUND: Comparative genomics aims to detect signals of evolutionary conservation as an indicator of functional constraint. Surprisingly, results of the ENCODE project revealed that about half of the experimentally verified functional elements found in non-coding DNA were classified as unconstrained by computational predictions. Following this observation, it has been hypothesized that this may be partly explained by biased estimates on neutral evolutionary rates used by existing sequence conservation metrics. All methods we are aware of rely on a comparison with the neutral rate and conservation is estimated by measuring the deviation of a particular genomic region from this rate. Consequently, it is a reasonable assumption that inaccurate neutral rate estimates may lead to biased conservation and constraint estimates. RESULTS: We propose a conservation signal that is produced by local Maximum Likelihood estimation of evolutionary parameters using an optimized sliding window and present a Kullback-Leibler projection that allows multiple different estimated parameters to be transformed into a conservation measure. This conservation measure does not rely on assumptions about neutral evolutionary substitution rates and little a priori assumptions on the properties of the conserved regions are imposed. We show the accuracy of our approach (KuLCons) on synthetic data and compare it to the scores generated by state-of-the-art methods (phastCons, GERP, SCONE) in an ENCODE region. We find that KuLCons is most often in agreement with the conservation/constraint signatures detected by GERP and SCONE while qualitatively very different patterns from phastCons are observed. Opposed to standard methods KuLCons can be extended to more complex evolutionary models, e.g. taking insertion and deletion events into account and corresponding results show that scores obtained under this model can diverge significantly from scores using the simpler model. CONCLUSION: Our results suggest that discriminating among the different degrees of conservation is possible without making assumptions about neutral rates. We find, however, that it cannot be expected to discover considerably different constraint regions than GERP and SCONE. Consequently, we conclude that the reported discrepancies between experimentally verified functional and computationally identified constraint elements are likely not to be explained by biased neutral rate estimates. PMID- 18405367 TI - Gene expression pattern of functional neuronal cells derived from human bone marrow mesenchymal stromal cells. AB - BACKGROUND: Neuronal tissue has limited potential to self-renew or repair after neurological diseases. Cellular therapies using stem cells are promising approaches for the treatment of neurological diseases. However, the clinical use of embryonic stem cells or foetal tissues is limited by ethical considerations and other scientific problems. Thus, bone marrow mesenchymal stomal cells (BM MSC) could represent an alternative source of stem cells for cell replacement therapies. Indeed, many studies have demonstrated that MSC can give rise to neuronal cells as well as many tissue-specific cell phenotypes. METHODS: BM-MSC were differentiated in neuron-like cells under specific induction (NPBM + cAMP + IBMX + NGF + Insulin). By day ten, differentiated cells presented an expression profile of real neurons. Functionality of these differentiated cells was evaluated by calcium influx through glutamate receptor AMPA3. RESULTS: Using microarray analysis, we compared gene expression profile of these different samples, before and after neurogenic differentiation. Among the 1943 genes differentially expressed, genes down-regulated are involved in osteogenesis, chondrogenesis, adipogenesis, myogenesis and extracellular matrix component (tuftelin, AGC1, FADS3, tropomyosin, fibronectin, ECM2, HAPLN1, vimentin). Interestingly, genes implicated in neurogenesis are increased. Most of them are involved in the synaptic transmission and long term potentialisation as cortactin, CASK, SYNCRIP, SYNTL4 and STX1. Other genes are involved in neurite outgrowth, early neuronal cell development, neuropeptide signaling/synthesis and neuronal receptor (FK506, ARHGAP6, CDKRAP2, PMCH, GFPT2, GRIA3, MCT6, BDNF, PENK, amphiregulin, neurofilament 3, Epha4, synaptotagmin). Using real time RT-PCR, we confirmed the expression of selected neuronal genes: NEGR1, GRIA3 (AMPA3), NEF3, PENK and Epha4. Functionality of these neuron-like cells was demonstrated by Ca2+ influx through glutamate receptor channel (AMPA3) in the presence of two agonist glutamate, AMPA or CNQX antagonist. CONCLUSION: Our results demonstrate that BM MSC have the potential to differentiate in neuronal cells with specific gene expression and functional properties. BM-MSC are thus promising candidates for cell-based therapy of neurodegenerative diseases. PMID- 18405368 TI - Surfactant proteins SP-B and SP-C and their precursors in bronchoalveolar lavages from children with acute and chronic inflammatory airway disease. AB - BACKGROUND: The surfactant proteins B (SP-B) and C (SP-C) are important for the stability and function of the alveolar surfactant film. Their involvement and down-regulation in inflammatory processes has recently been proposed, but their level during neutrophilic human airway diseases are not yet known. METHODS: We used 1D-electrophoresis and Western blotting to determine the concentrations and molecular forms of SP-B and SP-C in bronchoalveolar lavage (BAL) fluid of children with different inflammatory airway diseases. 21 children with cystic fibrosis, 15 with chronic bronchitis and 14 with pneumonia were included and compared to 14 healthy control children. RESULTS: SP-B was detected in BAL of all 64 patients, whereas SP-C was found in BAL of all but 3 children; those three BAL fluids had more than 80% neutrophils, and in two patients, who were re-lavaged later, SP-C was then present and the neutrophil count was lower. SP-B was mainly present as a dimer, SP-C as a monomer. For both qualitative and quantitative measures of SP-C and SP-B, no significant differences were observed between the four evaluated patient groups. CONCLUSION: Concentration or molecular form of SP B and SP-C is not altered in BAL of children with different acute and chronic inflammatory lung diseases. We conclude that there is no down-regulation of SP-B and SP-C at the protein level in inflammatory processes of neutrophilic airway disease. PMID- 18405370 TI - The effect of forearm posture on wrist flexion in computer workers with chronic upper extremity musculoskeletal disorders. AB - BACKGROUND: Occupational computer use has been associated with upper extremity musculoskeletal disorders (UEMSDs), but the etiology and pathophysiology of some of these disorders are poorly understood. Various theories attribute the symptoms to biomechanical and/or psychosocial stressors. The results of several clinical studies suggest that elevated antagonist muscle tension may be a biomechanical stress factor. Affected computer users often exhibit limited wrist range of motion, particularly wrist flexion, which has been attributed to increased extensor muscle tension, rather than to pain symptoms. Recreational or domestic activities requiring extremes of wrist flexion may produce injurious stress on the wrist joint and muscles, the symptoms of which are then exacerbated by computer use. As these activities may involve a variety of forearm postures, we examined whether changes in forearm posture have an effect on pain reports during wrist flexion, or whether pain would have a limiting effect on flexion angle. METHODS: We measured maximum active wrist flexion using a goniometer with the forearm supported in the prone, neutral, and supine postures. Data was obtained from 5 subjects with UEMSDs attributed to computer use and from 13 control subjects. RESULTS: The UEMSD group exhibited significantly restricted wrist flexion compared to the control group in both wrists at all forearm postures with the exception of the non-dominant wrist with the forearm prone. In both groups, maximum active wrist flexion decreased at the supine forearm posture compared to the prone posture. No UEMSD subjects reported an increase in pain symptoms during testing. CONCLUSION: The UEMSD group exhibited reduced wrist flexion compared to controls that did not appear to be pain related. A supine forearm posture reduced wrist flexion in both groups, but the reduction was approximately 100% greater in the UEMSD group. The effect of a supine forearm posture on wrist flexion is consistent with known biomechanical changes in the distal extensor carpi ulnaris tendon that occur with forearm supination. We infer from these results that wrist extensor muscle passive tension may be elevated in UEMSD subjects compared to controls, particularly in the extensor carpi ulnaris muscle. Measuring wrist flexion at the supine forearm posture may highlight flexion restrictions that are not otherwise apparent. PMID- 18405369 TI - Relationships between emm and multilocus sequence types within a global collection of Streptococcus pyogenes. AB - BACKGROUND: The M type-specific surface protein antigens encoded by the 5' end of emm genes are targets of protective host immunity and attractive vaccine candidates against infection by Streptococcus pyogenes, a global human pathogen. A history of genetic change in emm was evaluated for a worldwide collection of > 500 S. pyogenes isolates that were defined for genetic background by multilocus sequence typing of housekeeping genes. RESULTS: Organisms were categorized by genotypes that roughly correspond to throat specialists, skin specialists, and generalists often recovered from infections at either tissue site. Recovery of distant clones sharing the same emm type was approximately 4-fold higher for skin specialists and generalists, as compared to throat specialists. Importantly, emm type was often a poor marker for clone. Recovery of clones that underwent recombinational replacement with a new emm type was most evident for the throat and skin specialists. The average ratio of nonsynonymous substitutions per nonsynonymous site (Ka) and synonymous substitutions per synonymous site (Ks) was 4.9, 1.5 and 1.3 for emm types of the throat specialist, skin specialist and generalist groups, respectively. CONCLUSION: Data indicate that the relationships between emm type and genetic background differ among the three host tissue related groups, and that the selection pressures acting on emm appear to be strongest for the throat specialists. Since positive selection is likely due in part to a protective host immune response, the findings may have important implications for vaccine design and vaccination strategies. PMID- 18405371 TI - Apoptosis induction in Jurkat cells and sCD95 levels in women's sera are related with the risk of developing cervical cancer. AB - BACKGROUND: Currently, there is clear evidence that apoptosis plays an important role in the development and progression of tumors. One of the best characterized apoptosis triggering systems is the CD95/Fas/APO-1 pathway; previous reports have demonstrated high levels of soluble CD95 (sCD95) in serum of patients with some types of cancer. Cervical cancer is the second most common cancer among women worldwide. As a first step in an attempt to design a minimally invasive test to predict the risk of developing cervical cancer in patients with precancerous lesions, we used a simple assay based on the capacity of human serum to induce apoptosis in Jurkat cells. We evaluated the relationship between sCD95 levels and the ability to induce apoptosis in Jurkat cells in cervical cancer patients and controls. METHODS: Jurkat cells were exposed to serum from 63 women (20 healthy volunteers, 21 with cervical intraepithelial neoplasia grade I [CIN 1] and 22 with cervical-uterine carcinoma). The apoptotic rate was measured by flow cytometry using Annexin-V-Fluos and Propidium Iodide as markers. Serum levels of sCD95 and soluble CD95 ligand (sCD95L) were measured by ELISA kits. RESULTS: We found that serum from almost all healthy women induced apoptosis in Jurkat cells, while only fifty percent of the sera from women with CIN 1 induced cell death in Jurkat cells. Interestingly, only one serum sample from a patient with cervical uterine cancer was able to induce apoptosis, the rest of the sera protected Jurkat cells from this killing. We were able to demonstrate that elimination of Jurkat cells was mediated by the CD95/Fas/Apo-1 apoptotic pathway. Furthermore, the serum levels of sCD95 measured by ELISA were significantly higher in women with cervical cancer. CONCLUSION: Our results demonstrate that there is a strong correlation between low levels of sCD95 in serum of normal women and higher apoptosis induction in Jurkat cells. We suggest that an analysis of the apoptotic rate induced by serum in Jurkat cells and the levels of sCD95 in serum could be helpful during the prognosis and treatment of women detected with precancerous lesions or cervical cancer. PMID- 18405372 TI - Hepatotoxicity associated with sulfasalazine in inflammatory arthritis: A case series from a local surveillance of serious adverse events. AB - BACKGROUND: Spontaneous reporting systems for adverse drug reactions (ADRs) are handicapped by under-reporting and limited detail on individual cases. We report an investigation from a local surveillance for serious adverse drug reactions associated with disease modifying anti-rheumatic drugs that was triggered by the occurrence of liver failure in two of our patients. METHODS: Serious ADR reports have been solicited from local clinicians by regular postcards over the past seven years. Patients', who had hepatotoxicity on sulfasalazine and met a definition of a serious ADR, were identified. Two clinicians reviewed structured case reports and assessed causality by consensus and by using a causality assessment instrument. The likely frequency of hepatotoxicity with sulfasalazine was estimated by making a series of conservative assumptions. RESULTS: Ten cases were identified: eight occurred during surveillance. Eight patients were hospitalised, two in hepatic failure - one died after a liver transplant. All but one event occurred within 6 weeks of treatment. Seven patients had a skin rash, three eosinophilia and one interstitial nephritis. Five patients were of Black British of African or Caribbean descent. Liver enzymes showed a hepatocellular pattern in four cases and a mixed pattern in six. Drug-related hepatotoxicity was judged probable or highly probable in 8 patients. The likely frequency of serious hepatotoxicity with sulfasalazine was estimated at 0.4% of treated patients. CONCLUSION: Serious hepatotoxicity associated with sulfasalazine appears to be under-appreciated and intensive monitoring and vigilance in the first 6 weeks of treatment is especially important. PMID- 18405373 TI - Novel computational methods for increasing PCR primer design effectiveness in directed sequencing. AB - BACKGROUND: Polymerase chain reaction (PCR) is used in directed sequencing for the discovery of novel polymorphisms. As the first step in PCR directed sequencing, effective PCR primer design is crucial for obtaining high-quality sequence data for target regions. Since current computational primer design tools are not fully tuned with stable underlying laboratory protocols, researchers may still be forced to iteratively optimize protocols for failed amplifications after the primers have been ordered. Furthermore, potentially identifiable factors which contribute to PCR failures have yet to be elucidated. This inefficient approach to primer design is further intensified in a high-throughput laboratory, where hundreds of genes may be targeted in one experiment. RESULTS: We have developed a fully integrated computational PCR primer design pipeline that plays a key role in our high-throughput directed sequencing pipeline. Investigators may specify target regions defined through a rich set of descriptors, such as Ensembl accessions and arbitrary genomic coordinates. Primer pairs are then selected computationally to produce a minimal amplicon set capable of tiling across the specified target regions. As part of the tiling process, primer pairs are computationally screened to meet the criteria for success with one of two PCR amplification protocols. In the process of improving our sequencing success rate, which currently exceeds 95% for exons, we have discovered novel and accurate computational methods capable of identifying primers that may lead to PCR failures. We reveal the laboratory protocols and their associated, empirically determined computational parameters, as well as describe the novel computational methods which may benefit others in future primer design research. CONCLUSION: The high-throughput PCR primer design pipeline has been very successful in providing the basis for high-quality directed sequencing results and for minimizing costs associated with labor and reprocessing. The modular architecture of the primer design software has made it possible to readily integrate additional primer critique tests based on iterative feedback from the laboratory. As a result, the primer design software, coupled with the laboratory protocols, serves as a powerful tool for low and high-throughput primer design to enable successful directed sequencing. PMID- 18405374 TI - Comparative BAC end sequence analysis of tomato and potato reveals overrepresentation of specific gene families in potato. AB - BACKGROUND: Tomato (Solanum lycopersicon) and potato (S. tuberosum) are two economically important crop species, the genomes of which are currently being sequenced. This study presents a first genome-wide analysis of these two species, based on two large collections of BAC end sequences representing approximately 19% of the tomato genome and 10% of the potato genome. RESULTS: The tomato genome has a higher repeat content than the potato genome, primarily due to a higher number of retrotransposon insertions in the tomato genome. On the other hand, simple sequence repeats are more abundant in potato than in tomato. The two genomes also differ in the frequency distribution of SSR motifs. Based on EST and protein alignments, potato appears to contain up to 6,400 more putative coding regions than tomato. Major gene families such as cytochrome P450 mono-oxygenases and serine-threonine protein kinases are significantly overrepresented in potato, compared to tomato. Moreover, the P450 superfamily appears to have expanded spectacularly in both species compared to Arabidopsis thaliana, suggesting an expanded network of secondary metabolic pathways in the Solanaceae. Both tomato and potato appear to have a low level of microsynteny with A. thaliana. A higher degree of synteny was observed with Populus trichocarpa, specifically in the region between 15.2 and 19.4 Mb on P. trichocarpa chromosome 10. CONCLUSION: The findings in this paper present a first glimpse into the evolution of Solanaceous genomes, both within the family and relative to other plant species. When the complete genome sequences of these species become available, whole-genome comparisons and protein- or repeat-family specific studies may shed more light on the observations made here. PMID- 18405375 TI - uShuffle: a useful tool for shuffling biological sequences while preserving the k let counts. AB - BACKGROUND: Randomly shuffled sequences are routinely used in sequence analysis to evaluate the statistical significance of a biological sequence. In many cases, biologists need sophisticated shuffling tools that preserve not only the counts of distinct letters but also higher-order statistics such as doublet counts, triplet counts, and, in general, k-let counts. RESULTS: We present a sequence analysis tool (named uShuffle) for generating uniform random permutations of biological sequences (such as DNAs, RNAs, and proteins) that preserve the exact k let counts. The uShuffle tool implements the latest variant of the Euler algorithm and uses Wilson's algorithm in the crucial step of arborescence generation. It is carefully engineered and extremely efficient. The uShuffle tool achieves maximum flexibility by allowing arbitrary alphabet size and let size. It can be used as a command-line program, a web application, or a utility library. Source code in C, Java, and C#, and integration instructions for Perl and Python are provided. CONCLUSION: The uShuffle tool surpasses existing implementation of the Euler algorithm in both performance and flexibility. It is a useful tool for the bioinformatics community. PMID- 18405376 TI - Assessment of a non-invasive high-throughput classifier for behaviours associated with sleep and wake in mice. AB - This work presents a non-invasive high-throughput system for automatically detecting characteristic behaviours in mice over extended periods of time, useful for phenotyping experiments. The system classifies time intervals on the order of 2 to 4 seconds as corresponding to motions consistent with either active wake or inactivity associated with sleep. A single Polyvinylidine Difluoride (PVDF) sensor on the cage floor generates signals from motion resulting in pressure. This paper develops a linear classifier based on robust features extracted from normalized power spectra and autocorrelation functions, as well as novel features from the collapsed average (autocorrelation of complex spectrum), which characterize transient and periodic properties of the signal envelope. Performance is analyzed through an experiment comparing results from direct human observation and classification of the different behaviours with an automatic classifier used in conjunction with this system. Experimental results from over 28.5 hours of data from 4 mice indicate a 94% classification rate relative to the human observations. Examples of sequential classifications (2 second increments) over transition regions between sleep and wake behaviour are also presented to demonstrate robust performance to signal variation and explain performance limitations. PMID- 18405377 TI - Mental practice-based rehabilitation training to improve arm function and daily activity performance in stroke patients: a randomized clinical trial. AB - BACKGROUND: Over 50% of patients with upper limb paresis resulting from stroke face long-term impaired arm function and ensuing disability in daily life. Unfortunately, the number of effective treatments aimed at improving arm function due to stroke is still low. This study aims to evaluate a new therapy for improving arm function in sub-acute stroke patients based on mental practice theories and functional task-oriented training, and to study the predictors for a positive treatment result. It is hypothesized that a six-week, mental practice based training program (additional to regular therapy) targeting the specific upper extremity skills important to the individual patient will significantly improve both arm function and daily activity performance, as well as being cost effective. METHODS/DESIGN: One hundred and sixty sub-acute stroke patients with upper limb paresis (MRC grade 1-3) will participate in a single-blinded, multi centre RCT. The experimental group will undertake a six-week, individually tailored therapy regime focused on improving arm function using mental practice. The control group will perform bimanual upper extremity exercises in addition to regular therapy. Total contact time and training intensity will be similar for both groups. Measurements will be taken at therapy onset, after its cessation and during the follow-up period (after 6 and 12 months). Primary outcome measures will assess upper extremity functioning on the ICF level of daily life activity (Wolf Motor Function Test, Frenchay Arm Test, accelerometry), while secondary outcome measures cover the ICF impairment level (Brunnstrom-Fu-Meyer test). Level of societal participation (IPA) and quality of life (EuroQol; SS-Qol) will also be tested. Costs will be based on a cost questionnaire, and statistical analyses on MAN(C)OVA and GEE (generalized estimated equations). DISCUSSION: The results of this study will provide evidence on the effectiveness of this mental practice based rehabilitation training, as well as the cost-effectiveness. TRIAL REGISTRATION: Current Controlled Trials [ISRCTN33487341). PMID- 18405379 TI - Phylogenetic distribution of large-scale genome patchiness. AB - BACKGROUND: The phylogenetic distribution of large-scale genome structure (i.e. mosaic compositional patchiness) has been explored mainly by analytical ultracentrifugation of bulk DNA. However, with the availability of large, good quality chromosome sequences, and the recently developed computational methods to directly analyze patchiness on the genome sequence, an evolutionary comparative analysis can be carried out at the sequence level. RESULTS: The local variations in the scaling exponent of the Detrended Fluctuation Analysis are used here to analyze large-scale genome structure and directly uncover the characteristic scales present in genome sequences. Furthermore, through shuffling experiments of selected genome regions, computationally-identified, isochore-like regions were identified as the biological source for the uncovered large-scale genome structure. The phylogenetic distribution of short- and large-scale patchiness was determined in the best-sequenced genome assemblies from eleven eukaryotic genomes: mammals (Homo sapiens, Pan troglodytes, Mus musculus, Rattus norvegicus, and Canis familiaris), birds (Gallus gallus), fishes (Danio rerio), invertebrates (Drosophila melanogaster and Caenorhabditis elegans), plants (Arabidopsis thaliana) and yeasts (Saccharomyces cerevisiae). We found large-scale patchiness of genome structure, associated with in silico determined, isochore-like regions, throughout this wide phylogenetic range. CONCLUSION: Large-scale genome structure is detected by directly analyzing DNA sequences in a wide range of eukaryotic chromosome sequences, from human to yeast. In all these genomes, large-scale patchiness can be associated with the isochore-like regions, as directly detected in silico at the sequence level. PMID- 18405378 TI - Limited functional conservation of a global regulator among related bacterial genera: Lrp in Escherichia, Proteus and Vibrio. AB - BACKGROUND: Bacterial genome sequences are being determined rapidly, but few species are physiologically well characterized. Predicting regulation from genome sequences usually involves extrapolation from better-studied bacteria, using the hypothesis that a conserved regulator, conserved target gene, and predicted regulator-binding site in the target promoter imply conserved regulation between the two species. However many compared organisms are ecologically and physiologically diverse, and the limits of extrapolation have not been well tested. In E. coli K-12 the leucine-responsive regulatory protein (Lrp) affects expression of approximately 400 genes. Proteus mirabilis and Vibrio cholerae have highly-conserved lrp orthologs (98% and 92% identity to E. coli lrp). The functional equivalence of Lrp from these related species was assessed. RESULTS: Heterologous Lrp regulated gltB, livK and lrp transcriptional fusions in an E. coli background in the same general way as the native Lrp, though with significant differences in extent. Microarray analysis of these strains revealed that the heterologous Lrp proteins significantly influence only about half of the genes affected by native Lrp. In P. mirabilis, heterologous Lrp restored swarming, though with some pattern differences. P. mirabilis produced substantially more Lrp than E. coli or V. cholerae under some conditions. Lrp regulation of target gene orthologs differed among the three native hosts. Strikingly, while Lrp negatively regulates its own gene in E. coli, and was shown to do so even more strongly in P. mirabilis, Lrp appears to activate its own gene in V. cholerae. CONCLUSION: The overall similarity of regulatory effects of the Lrp orthologs supports the use of extrapolation between related strains for general purposes. However this study also revealed intrinsic differences even between orthologous regulators sharing >90% overall identity, and 100% identity for the DNA-binding helix-turn-helix motif, as well as differences in the amounts of those regulators. These results suggest that predicting regulation of specific target genes based on genome sequence comparisons alone should be done on a conservative basis. PMID- 18405380 TI - Consistent and contrasting properties of lineage-specific genes in the apicomplexan parasites Plasmodium and Theileria. AB - BACKGROUND: Lineage-specific genes, the genes that are restricted to a limited subset of related organisms, may be important in adaptation. In parasitic organisms, lineage-specific gene products are possible targets for vaccine development or therapeutics when these genes are absent from the host genome. RESULTS: In this study, we utilized comparative approaches based on a phylogenetic framework to characterize lineage-specific genes in the parasitic protozoan phylum Apicomplexa. Genes from species in two major apicomplexan genera, Plasmodium and Theileria, were categorized into six levels of lineage specificity based on a nine-species phylogeny. In both genera, lineage-specific genes tend to have a higher level of sequence divergence among sister species. In addition, species-specific genes possess a strong codon usage bias compared to other genes in the genome. We found that a large number of genus- or species specific genes are putative surface antigens that may be involved in host parasite interactions. Interestingly, the two parasite lineages exhibit several notable differences. In Plasmodium, the (G + C) content at the third codon position increases with lineage specificity while Theileria shows the opposite trend. Surface antigens in Plasmodium are species-specific and mainly located in sub-telomeric regions. In contrast, surface antigens in Theileria are conserved at the genus level and distributed across the entire lengths of chromosomes. CONCLUSION: Our results provide further support for the model that gene duplication followed by rapid divergence is a major mechanism for generating lineage-specific genes. The result that many lineage-specific genes are putative surface antigens supports the hypothesis that lineage-specific genes could be important in parasite adaptation. The contrasting properties between the lineage specific genes in two major apicomplexan genera indicate that the mechanisms of generating lineage-specific genes and the subsequent evolutionary fates can differ between related parasite lineages. Future studies that focus on improving functional annotation of parasite genomes and collection of genetic variation data at within- and between-species levels will be important in facilitating our understanding of parasite adaptation and natural selection. PMID- 18405381 TI - Cytokine generation, promoter activation, and oxidant-independent NF-kappaB activation in a transfectable human neutrophilic cellular model. AB - BACKGROUND: Human neutrophils are key players of innate immunity, and influence inflammatory and immune reactions through the production of numerous cytokines and chemokines. Despite major advances in our understanding of this important functional response of neutrophils, the short lifespan of these cells and their resistance to transfection have always been an obstacle to the detailed dissection of signaling pathways and effector responses that is often possible in other cell types. RESULTS: Here, we report that granulocytic differentiation of human PLB-985 cells with DMSO yields cells that are neutrophil-like with respect to surface markers, acquisition of responsiveness to physiological neutrophil stimuli (fMLP, LPS), cytokine expression and production profile, and transcription factor activation profile (NF-kappaB, C/EBP, AP-1, STAT). We also show that granulocytic PLB-985 cells can be reliably tranfected by nucleofection in a rapid and efficient manner. Indeed, we overexpressed several proteins and luciferase constructs into these cells. In particular, overexpression of a dominant negative IkappaB-alpha confirmed the central role of NF-kappaB in the production of cytokines by granulocytes. Moreover, the use of PLB-985 granulocytes in which the NADPH oxidase is inactive due to the targeted disruption of a key component (gp91phox) revealed that NF-kappaB activation and kappaB-dependent responses are independent of endogenous reactive oxygen intermediates in these cells. Antioxidant studies performed in primary human neutrophils support this conclusion. CONCLUSION: Our results unveil a new facet of the NF-kappaB system of human granulocytes, and pave the way for deciphering signal transduction pathways and promoter activation in these cells. PMID- 18405382 TI - Five-Factor Model personality profiles of drug users. AB - BACKGROUND: Personality traits are considered risk factors for drug use, and, in turn, the psychoactive substances impact individuals' traits. Furthermore, there is increasing interest in developing treatment approaches that match an individual's personality profile. To advance our knowledge of the role of individual differences in drug use, the present study compares the personality profile of tobacco, marijuana, cocaine, and heroin users and non-users using the wide spectrum Five-Factor Model (FFM) of personality in a diverse community sample. METHOD: Participants (N = 1,102; mean age = 57) were part of the Epidemiologic Catchment Area (ECA) program in Baltimore, MD, USA. The sample was drawn from a community with a wide range of socio-economic conditions. Personality traits were assessed with the Revised NEO Personality Inventory (NEO PI-R), and psychoactive substance use was assessed with systematic interview. RESULTS: Compared to never smokers, current cigarette smokers score lower on Conscientiousness and higher on Neuroticism. Similar, but more extreme, is the profile of cocaine/heroin users, which score very high on Neuroticism, especially Vulnerability, and very low on Conscientiousness, particularly Competence, Achievement-Striving, and Deliberation. By contrast, marijuana users score high on Openness to Experience, average on Neuroticism, but low on Agreeableness and Conscientiousness. CONCLUSION: In addition to confirming high levels of negative affect and impulsive traits, this study highlights the links between drug use and low Conscientiousness. These links provide insight into the etiology of drug use and have implications for public health interventions. PMID- 18405383 TI - The evolutionary trajectory of the mating-type (mat) genes in Neurospora relates to reproductive behavior of taxa. AB - BACKGROUND: Comparative sequencing studies among a wide range of taxonomic groups, including fungi, have led to the discovery that reproductive genes evolve more rapidly than other genes. However, for fungal reproductive genes the question has remained whether the rapid evolution is a result of stochastic or deterministic processes. The mating-type (mat) genes constitute the master regulators of sexual reproduction in filamentous ascomycetes and here we present a study of the molecular evolution of the four mat-genes (mat a-1, mat A-1, mat A 2 and mat A-3) of 20 Neurospora taxa. RESULTS: We estimated nonsynonymous and synonymous substitution rates of genes to infer their evolutionary rate, and confirmed that the mat-genes evolve rapidly. Furthermore, the evolutionary trajectories are related to the reproductive modes of the taxa; likelihood methods revealed that positive selection acting on specific codons drives the diversity in heterothallic taxa, while among homothallic taxa the rapid evolution is due to a lack of selective constraint. The latter finding is supported by presence of stop codons and frame shift mutations disrupting the open reading frames of mat a-1, mat A-2 and mat A-3 in homothallic taxa. Lower selective constraints of mat-genes was found among homothallic than heterothallic taxa, and comparisons with non-reproductive genes argue that this disparity is not a nonspecific, genome-wide phenomenon. CONCLUSION: Our data show that the mat-genes evolve rapidly in Neurospora. The rapid divergence is due to either adaptive evolution or lack of selective constraints, depending on the reproductive mode of the taxa. This is the first instance of positive selection acting on reproductive genes in the fungal kingdom, and illustrates how the evolutionary trajectory of reproductive genes can change after a switch in reproductive behaviour of an organism. PMID- 18405384 TI - The PI3K pathway regulates endochondral bone growth through control of hypertrophic chondrocyte differentiation. AB - BACKGROUND: The majority of our bones develop through the process of endochondral ossification that involves chondrocyte proliferation and hypertrophic differentiation in the cartilage growth plate. A large number of growth factors and hormones have been implicated in the regulation of growth plate biology, however, less is known about the intracellular signaling pathways involved. PI3K/Akt has been identified as a major regulator of cellular proliferation, differentiation and death in multiple cell types. RESULTS AND DISCUSSION: Employing an organ culture system of embryonic mouse tibiae and LY294002, a pharmacological inhibitor of PI3K, we show that inhibition of the pathway results in significant growth reduction, demonstrating that PI3K is required for normal endochondral bone growth in vitro. PI3K inhibition reduces the length of the proliferating and particularly of the hypertrophic zone. Studies with organ cultures and primary chondrocytes in micromass culture show delayed hypertrophic differentiation of chondrocytes and increased apoptosis in the presence of LY294002. Surprisingly, PI3K inhibition had no strong effect on IGF1-induced bone growth, but partially blocked the anabolic effects of C-type natriuretic peptide. CONCLUSION: Our data demonstrate an essential role of PI3K signaling in chondrocyte differentiation and as a consequence of this, in the endochondral bone growth process. PMID- 18405385 TI - The influences of Taiwan's generic grouping price policy on drug prices and expenditures: evidence from analysing the consumption of the three most-used classes of cardiovascular drugs. AB - BACKGROUND: Controlling the growth of pharmaceutical expenditures is a major global challenge. Promotion of generic drug prescriptions or use is gaining increased support. There are substantial contextual differences in international experiences of implementing pharmaceutical policies related to generic drugs. Reporting these experiences from varied perspectives can inform future policy making. This study describes an experience of Taiwan, where patients with chronic (long-term) conditions are usually managed in hospitals and drugs are provided in this setting with costs reimbursed through the National Health Insurance (NHI). It investigates the effects of Taiwan's reimbursement rate adjustment based on chemical generic grouping in 2001. This research also demonstrates the use of micro-level longitudinal data to generate policy-relevant information. The research can be used to improve efficiency of health care resource use. METHODS: We chose the three most-used classes of cardiovascular drugs for this investigation: beta blocking agents, calcium channel blockers mainly with vascular effects, and plain ACE inhibitors. For each drug class, we investigated changes in daily expense, consumption volume, and total expenditures from a pre action period to a corresponding post-action period. We compared an exposure or "intervention" group of patients targeted by the action with a comparisonor "control" group of patients not targeted by the action. The data sources are a longitudinal database for 200,000 NHI enrolees, corresponding NHI registration data of health care facilities, and an archive recording all historical data on the reimbursement rates of drugs covered by the NHI. We adopted a fixed effects linear regression model to control for unobserved heterogeneity among patient hospital groups. Additional descriptive statistics were applied to examine whether any inappropriate consumption of drugs in the three classes existed. RESULTS: The daily drug expense significantly decreased from the pre-action period to the post-action period for the exposure group. The average magnitudes of the decreases for the three classes of drugs mentioned above were 14.8%, 5.8% and 5.8%, respectively. In contrast, there was no reduction for the comparison group. The number of days of the prescription increased significantly from the pre- to the post-action period for both exposure and comparison groups. The total expense also significantly increased for both patient groups. For the exposure group, the average magnitudes of the growth in the total expenditure for the three classes of drugs were 47.7%, 60.0% and 55.3%, respectively. For the comparison group, they were 91.6%, 91.6% and 63.2%, respectively. After the action, approximately 50% of patients obtained more than 180 days of prescription drugs for a six-month period. CONCLUSION: The 2001 price adjustment action, based on generic grouping, significantly reduced the daily expense of each of the three classes of cardiovascular drugs. However, in response to this policy change, hospitals in Taiwan tended to greatly expand the volume of drugs prescribed for their regular patients. Consequently, the total expenditures for the three classes of drugs grew substantially after the action. These knock-on effects weakened the capability of the price adjustment action to control total pharmaceutical expenditures. This means that no saved resources were available for other health care uses. Such expansion of pharmaceutical consumption might also lead to inefficient use of the three drug classes: a large proportion of patients obtained more than one day of drugs per day in the post-action period, suggesting manipulation to increase reimbursement and offset price controls. We recommend that Taiwan's government use the NHI data to establish a monitoring system to detect inappropriate prescription patterns before implementing future policy changes. Such a monitoring system could then be used to deter hospitals from abusing their prescription volumes, making it possible to more effectively save health care resources by reducing drug reimbursement rates. PMID- 18405386 TI - Different tissue reaction of oesophagus and diaphragm after mesh hiatoplasty. Results of an animal study. AB - BACKGROUND: Laparoscopic mesh-reinforcement of the hiatal region in the treatment of gastroesophageal reflux disease (GERD) and paraesophageal hernia (PEH) reduces the risk of recurrence. However, there are still controversies about the technique of mesh placement, shape, structure and material. We therefore compared tissue integration and scar formation after implantation of two different polypropylene-meshes in a rabbit model. METHODS: A total of 20 female chinchilla rabbits were included in this study. Two different meshes (Polypropylene PP, Polyglecaprone 25 Composite PP-PG) were implanted on the abdominal diaphragm around the oesophagus. After 3 months the implanted meshes were excised en-bloc. Histological and morphological analyses were carried out accordingly proliferation rate, apoptosis and collagen type I/III ratio. RESULTS: Regarding proliferation rate of oesophagus PP (9.31 +/- 3.4%) and PP-PG (13.26 +/- 2.54%) differ in a significant (p = 0.0097) way. In the diaphragm we found a significant (p = 0.00066) difference between PP (9.43 +/- 1.45%) and PP-PG (18.73 +/- 5.92%) respectively. Comparing oesophagus and diaphragm we could prove a significant difference within PP-PG-group (p = 0.0195). Within PP-group the difference reached no statistical significance (p = 0.88). We found analogous results regarding apoptosis.Furthermore, there is a significant (p = 0.00013) difference of collagen type I/III ratio in PP-PG (12.28 +/- 0.8) compared to PP (8.44 +/- 1,63) in case of oesophageal tissue. Concerning diaphragm we found a significant difference (p = 0.000099) between PP-PG (8.85 +/- 0.81) and PP (6.32 +/- 1.07) as well. CONCLUSION: The histologic and morphologic characteristics after prosthetic enforcement of the hiatus in this animal model show a more distinct tissue integration using PP-PG compared to PP. Additionally, different wound healing and remodelling capability influence tissue integration of the mesh in diaphragm and oesophagus. PMID- 18405387 TI - The genome of the brown alga Ectocarpus siliculosus contains a series of viral DNA pieces, suggesting an ancient association with large dsDNA viruses. AB - BACKGROUND: Ectocarpus siliculosus virus-1 (EsV-1) is a lysogenic dsDNA virus belonging to the super family of nucleocytoplasmic large DNA viruses (NCLDV) that infect Ectocarpus siliculosus, a marine filamentous brown alga. Previous studies indicated that the viral genome is integrated into the host DNA. In order to find the integration sites of the viral genome, a genomic library from EsV-1-infected algae was screened using labelled EsV-1 DNA. Several fragments were isolated and some of them were sequenced and analyzed in detail. RESULTS: Analysis revealed that the algal genome is split by a copy of viral sequences that have a high identity to EsV-1 DNA sequences. These fragments are interspersed with DNA repeats, pseudogenes and genes coding for products involved in DNA replication, integration and transposition. Some of these gene products are not encoded by EsV 1 but are present in the genome of other members of the NCLDV family. Further analysis suggests that the Ectocarpus algal genome contains traces of the integration of a large dsDNA viral genome; this genome could be the ancestor of the extant NCLDV genomes. Furthermore, several lines of evidence indicate that the EsV-1 genome might have originated in these viral DNA pieces, implying the existence of a complex integration and recombination system. A protein similar to a new class of tyrosine recombinases might be a key enzyme of this system. CONCLUSION: Our results support the hypothesis that some dsDNA viruses are monophyletic and evolved principally through genome reduction. Moreover, we hypothesize that phaeoviruses have probably developed an original replication system. PMID- 18405388 TI - Range of shoulder motion in patients with adhesive capsulitis; intra-tester reproducibility is acceptable for group comparisons. AB - BACKGROUND: Measurements of range of motion play a key role in shoulder research. The purpose of this study is to investigate intra-observer reproducibility of measurements of active and passive range of motion in patients with adhesive capsulitis. METHODS: The study was carried out in a population consisting of 32 patients with clinical signs of adhesive capsulitis. A specified measurement protocol was used, and range of motion in affected and non-affected shoulders was measured twice for each patient with a one-week interval. RESULTS: For most of the investigated individual movements, test-retest differences in range of motion score of more than approximately 15 degrees are not likely to occur as a result of measurement error only. Point-estimates for the intraclass correlation coefficient ranged from 0.61 to 0.93. CONCLUSION: Range of motion of patients with adhesive capsulitis can be measured with acceptable reproducibility in settings where groups are compared. Scores for individual patients should be interpreted with caution. PMID- 18405390 TI - A randomized double-blind placebo-controlled trial to investigate the effects of nasal calcitonin on bone microarchitecture measured by high-resolution peripheral quantitative computerized tomography in postmenopausal women - study protocol. AB - BACKGROUND: Bone microarchitecture is a significant determinant of bone strength. So far, the assessment of bone microarchitecture has required bone biopsies, limiting its utilization in clinical practice to one single skeletal site. With the advance of high-resolution imaging techniques, non-invasive in vivo measurement of bone microarchitecture has recently become possible. This provides an opportunity to efficiently assess the effects of anti-osteoporotic therapies on bone microarchitecture. We therefore designed a protocol to investigate the effects of nasal salmon calcitonin, an inhibitor of osteoclast activity, on bone microarchitecture in postmenopausal women, comparing weight bearing and non weight bearing skeletal sites. METHODS: One hundred postmenopausal women will be included in a randomized, placebo-controlled, double-blind trial comparing the effect of nasal salmon calcitonin (200 UI/day) to placebo over two years. Bone microarchitecture at the distal radius and distal tibia will be determined yearly by high-resolution peripheral quantitative computerized tomography (p-QCT) with a voxel size of 82 mum and an irradiation of less than 5 muSv. Serum markers of bone resorption and bone formation will be measured every 6 months. Safety and compliance will be assessed. Primary endpoint is the change in bone microarchitecture; secondary endpoint is the change in markers of bone turnover. HYPOTHESIS: The present study should provide new information on the mode of action of nasal calcitonin. We hypothezise that - compared to placebo - calcitonin impacts on microstructural parameters, with a possible difference between weight bearing and non-weight bearing bones. TRIAL REGISTRATION: ClinicalTrials.gov NCT00372099. PMID- 18405389 TI - Low prevalence of Chlamydia trachomatis infection in asymptomatic young Swiss men. AB - BACKGROUND: Prevalence and risk factors for Chlamydia trachomatis infection among young men in Switzerland is still unknown. The objective of the present study was to assess prevalence and risk factors for C. trachomatis infection in young Swiss men. METHODS: 517 young Swiss men were enrolled in this cross-sectional study during their compulsory military recruitment. Participants completed a questionnaire and gave urine samples which were screened for C. trachomatis DNA by PCR. Genotyping of positive samples was done by amplification and sequencing the ompA gene. RESULTS: The prevalence of chlamydial infection among young Swiss male was 1.2% (95% confidence interval [95%CI], 0.4-2.5%). C. trachomatis infection was only identified among the 306 men having multiple sexual partner. Although frequent, neither unprotected sex (absence of condom use), nor alcohol and drug abuse were associated with chlamydial infection. Men living in cities were more frequently infected (2.9%, 95%CI 0.8-7.4%) than men living in rural areas (0.5%, 95%CI 0.1-1.9%, p = 0.046). Moreover, naturalised Swiss citizens were more often positive (4.9%, 95%CI 1.3-12.5%) than native-born Swiss men (0.5%, 95%CI 0.1-1.7%, p = 0.003). CONCLUSION: In comparison with other countries, the prevalence of chlamydial infection in men is extremely low in Switzerland, despite a significant prevalence of risky sexual behaviour. C. trachomatis infection was especially prevalent in men with multiple sexual partners. Further research is required (i) to define which subgroup of the general population should be routinely screened, and (ii) to test whether such a targeted screening strategy will be effective to reduce the prevalence of chlamydial infection among this population. PMID- 18405391 TI - Expression of estrogen receptor beta in the breast carcinoma of BRCA1 mutation carriers. AB - BACKGROUND: Breast cancers (BC) in women carrying mutations in BRCA1 gene are more frequently estrogen receptor negative than the nonhereditary BC. Nevertheless, tamoxifen has been found to have a protective effect in preventing contralateral tumors in BRCA1 mutation carriers. The identification of the second human estrogen receptor, ERbeta, raised a question of its role in hereditary breast cancer. The aim of this study was to assess the frequency of ERalpha, ERbeta, PgR (progesterone receptor) and HER-2 expression in breast cancer patients with mutated BRCA1 gene and in the control group. METHODS: The study group consisted of 48 women with BRCA1 gene mutations confirmed by multiplex PCR assay. The patients were tested for three most common mutations of BRCA1 affecting the Polish population (5382insC, C61G, 4153delA). Immunostaining for ERalpha, ERbeta and PgR (progesterone receptor) was performed using monoclonal antibodies against ERalpha, PgR (DakoCytomation), and polyclonal antibody against ERbeta (Chemicon). The EnVision detection system was applied. The study population comprised a control group of 120 BC operated successively during the years 1998-99. RESULTS: The results of our investigation showed that BRCA1 mutation carriers were more likely to have ERalpha-negative breast cancer than those in the control group. Only 14.5% of BRCA1-related cancers were ERalpha positive compared with 57.5% in the control group (P < 0.0001). On the contrary, the expression of ERbeta protein was observed in 42% of BRCA1-related tumors and in 55% of the control group. An interesting finding was that most hereditary cancers (75% of the whole group) were triple-negative: ERalpha(-)/PgR(-)/HER-2(-) but almost half of this group (44.4%) showed the expression of ERbeta. CONCLUSION: In the case of BRCA1-associated tumors the expression of ERbeta was significantly higher than the expression of ERalpha. This may explain the effectiveness of tamoxifen in preventing contralateral breast cancer development in BRCA1 mutation carriers. PMID- 18405392 TI - Genome-wide search reveals a novel GacA-regulated small RNA in Pseudomonas species. AB - BACKGROUND: Small RNAs (sRNAs) are widespread among bacteria and have diverse regulatory roles. Most of these sRNAs have been discovered by a combination of computational and experimental methods. In Pseudomonas aeruginosa, a ubiquitous Gram-negative bacterium and opportunistic human pathogen, the GacS/GacA two component system positively controls the transcription of two sRNAs (RsmY, RsmZ), which are crucial for the expression of genes involved in virulence. In the biocontrol bacterium Pseudomonas fluorescens CHA0, three GacA-controlled sRNAs (RsmX, RsmY, RsmZ) regulate the response to oxidative stress and the expression of extracellular products including biocontrol factors. RsmX, RsmY and RsmZ contain multiple unpaired GGA motifs and control the expression of target mRNAs at the translational level, by sequestration of translational repressor proteins of the RsmA family. RESULTS: A combined computational and experimental approach enabled us to identify 14 intergenic regions encoding sRNAs in P. aeruginosa. Eight of these regions encode newly identified sRNAs. The intergenic region 1698 was found to specify a novel GacA-controlled sRNA termed RgsA. GacA regulation appeared to be indirect. In P. fluorescens CHA0, an RgsA homolog was also expressed under positive GacA control. This 120-nt sRNA contained a single GGA motif and, unlike RsmX, RsmY and RsmZ, was unable to derepress translation of the hcnA gene (involved in the biosynthesis of the biocontrol factor hydrogen cyanide), but contributed to the bacterium's resistance to hydrogen peroxide. In both P. aeruginosa and P. fluorescens the stress sigma factor RpoS was essential for RgsA expression. CONCLUSION: The discovery of an additional sRNA expressed under GacA control in two Pseudomonas species highlights the complexity of this global regulatory system and suggests that the mode of action of GacA control may be more elaborate than previously suspected. Our results also confirm that several GGA motifs are required in an sRNA for sequestration of the RsmA protein. PMID- 18405393 TI - Reducing stigma and discrimination: Candidate interventions. AB - This paper proposes that stigma in relation to people with mental illness can be understood as a combination of problems of knowledge (ignorance), attitudes (prejudice) and behaviour (discrimination). From a literature review, a series of candidate interventions are identified which may be effective in reducing stigmatisation and discrimination at the following levels: individuals with mental illness and their family members; the workplace; and local, national and international. The strongest evidence for effective interventions at present is for (i) direct social contact with people with mental illness at the individual level, and (ii) social marketing at the population level. PMID- 18405394 TI - Survey of patient satisfaction with the Breastfeeding Education and Support Services of The Royal Women's Hospital, Melbourne. AB - BACKGROUND: The Breastfeeding Education and Support Services (BESS) is a unit of The Royal Women's Hospital in Melbourne, Australia, staffed by International Board Certified Lactation Consultants (IBCLCs), providing day/short-stay and an outpatient clinic for mothers and infants with breastfeeding problems. It is important to measure women's experience of visiting the service as part of quality assurance. The aim of this project was to conduct an anonymous postal survey of clients' satisfaction with BESS. METHODS: An anonymous survey was posted on 16 November 2005 and again on 31 January 2006, to all women who had attended BESS in September 2005. RESULTS: The response rate was 60.5% (78/129). Eighty percent (62/78) of respondents attended day-stay, 33% (26/78) attended short-stay and 15% (12/78) attended the outpatient clinic. The percentage of women who responded "strongly agree" to the statement "Overall, I am satisfied with the services" was 49% (35/72) and 50% (6/12) for those who went to day/short stay and the outpatient clinic respectively. Overall, 56% of all respondents responded that the quality of BESS was "better than expected". The most common breastfeeding problem reported was difficulty attaching the baby to the breast, followed by nipple damage, low milk supply and painful feeding. CONCLUSION: BESS seems to have provided a satisfactory service to most clients. Most respondents were clearly satisfied with the support given by the IBCLCs and have also responded that the staff were professional and knowledgeable in their field of work. PMID- 18405396 TI - Ageism among Israeli students: structure and demographic influences. AB - BACKGROUND: Research concerning negative attitudes toward the elderly among samples of North American respondents has uncovered two distinct yet comparable three-factor structures: (1) avoidance, antilocution and separation; and (2) discrimination, stereotypes and affective attitudes. In addition, previous research has demonstrated that men and younger people have more negative attitudes toward the elderly than do women and older people. The aim of this study is to examine the extent to which these findings can be generalized to individuals from a different culture. METHOD: The Fraboni Scale of Ageism (FSA) was administered to 491 Israeli participants aged 20-50 years. RESULTS: Exploratory factor analysis with varimax rotation uncovered a three-factor structure, similar to previous investigations. In contrast to the literature, however, no simple group (gender, age) differences were found for ageism. However, discriminant function analysis revealed a more complex relationship between age, gender and ageism. Older males, in contrast to older females, demonstrated a tendency to perceptions of the elderly that associated avoidance of the elderly with negative stereotypes whereas older females, in contrast to younger females, held perceptions that associated avoidance of the elderly with a negative perception of the elderly person's contribution to society. CONCLUSIONS: Despite cultural differences between Israel and North America, the similarity of findings provides support for the generalizabilty of the structure of attitudes toward the elderly as measured by the FSA to various cultures. In addition, the combined effect of age and gender on differential forms of ageism is complex and should be examined in future research. PMID- 18405395 TI - A novel c.5308_5311delGAGA mutation in Senataxin in a Cypriot family with an autosomal recessive cerebellar ataxia. AB - BACKGROUND: Senataxin (chromosome 9q34) was recently identified as the causative gene for an autosomal recessive form of Ataxia (ARCA), termed as Ataxia with Oculomotor Apraxia, type 2 (AOA2) and characterized by generalized incoordination, cerebellar atrophy, peripheral neuropathy, "oculomotor apraxia" and increased alpha-fetoprotein (AFP). Here, we report a novel Senataxin mutation in a Cypriot ARCA family. METHODS: We studied several Cypriot autosomal recessive cerebellar ataxia (ARCA) families for linkage to known ARCA gene loci. We linked one family (909) to the SETX locus on chromosome 9q34 and screened the proband for mutations by direct sequencing. RESULTS: Sequence analysis revealed a novel c.5308_5311delGAGA mutation in exon 11 of the SETX gene. The mutation has not been detected in 204 control chromosomes from the Cypriot population, the remaining Cypriot ARCA families and 37 Cypriot sporadic cerebellar ataxia patients. CONCLUSION: We identified a novel SETX homozygous c.5308_5311delGAGA mutation that co-segregates with ARCA with cerebellar atrophy and raised AFP. PMID- 18405397 TI - Knowledge and attitudes of doctors toward the sexuality of older people in Turkey. AB - BACKGROUND: Few studies have looked at healthcare professionals' knowledge of and attitudes to later life sexuality in both Western and Eastern cultures. Here we examine the attitudes and knowledge of Turkish medical doctors toward sexuality in older people. METHOD: Eighty-seven doctors, from various specialties, who were directly involved in the care of older people, were contacted by post and asked to complete the Turkish version of the Aging Sexual Knowledge and Attitudes Scale (ASKAS). RESULTS: A majority of physicians indicated that they had limited information and knowledge regarding sexual health issues in older people (69%). Although a small percentage (14.5%) reported that they "always" discuss sexuality and sexual problems with older patients, the majority (69%) indicated that they "sometimes" raise questions about sexuality with these patients. A high percentage (81%) stated that they would be helpful and receptive should an elderly patient initiate a discussion about sexual issues. Most participants (77%) thought that the patient's gender was of no importance when taking a sexual history. Overall, the responses to ASKAS showed that physicians had limited knowledge but their attitude was positive toward sexuality in the elderly. Female physicians had less knowledge than males and had more negative attitudes toward sexuality in this age group. Total and knowledge subscale scores of ASKAS showed that older physicians had more knowledge than younger physicians but similar attitudes. A comparison of the knowledge and attitude scores of psychiatrists, surgeons and non-surgeons showed no significant difference among the three groups. CONCLUSION: This study identified a low level of awareness of later life sexuality among Turkish medical doctors. These findings identify a need to improve the education and training of doctors at both undergraduate and postgraduate levels to enable them to provide better sexual health care to older people. PMID- 18405398 TI - Short-term practice effects in amnestic mild cognitive impairment: implications for diagnosis and treatment. AB - BACKGROUND: Practice effects have been widely reported in healthy older adults, but these improvements due to repeat exposure to test materials have been more equivocal in individuals with mild cognitive impairment (MCI). METHODS: The current study examined short-term practice effects in MCI by repeating a brief battery of cognitive tests across one week in 59 older adults with amnestic MCI and 62 intact older adults. RESULTS: Participants with amnestic MCI showed significantly greater improvements on two delayed recall measures (p < 0.01) compared to intact peers. All other practice effects were comparable between these two groups. Practice effects significantly improved scores in the MCI group so that 49% of them were reclassified as "intact" after one week, whereas the other 51% remained "stable" as MCI. Secondary analyses indicated the MCI-Intact group demonstrated larger practice effects on two memory measures than their peers (p < 0.01). CONCLUSIONS: These results continue to inform us about the nature of memory deficits in MCI, and could have implications for the diagnosis and possible treatment of this amnestic condition. PMID- 18405399 TI - Decreased initiation of usual activities in people with mild-to-moderate Alzheimer's disease: a descriptive analysis from the VISTA clinical trial. AB - BACKGROUND: Decreased initiation of activities is a common symptom in Alzheimer's disease (AD) and creates significant burden for carers, yet little is known about how patients, carers and treating physicians describe this problem. METHODS: This is a secondary, qualitative analysis of video-recorded, semi-structured and open ended interviews with the 130 community-dwelling patients with mild-to-moderate AD from the Video-Imaging Synthesis of Treating Alzheimer's Disease (VISTA) study - a four-month, randomized, placebo-controlled trial of galantamine. Interviews were coded and organized using Atlas.ti. By applying a framework analysis approach, patient/carer descriptions of dementia-related decreased initiation of usual tasks and activities were categorized and summarized. RESULTS: Decreased initiation of instrumental activities of daily living (IADLs) and/or leisure and social activities was reported in 85/130 patients at baseline, 71 (84%) of whom specifically targeted increased initiation as a goal of treatment. The most common manifestations of decreased initiation were initiating only with prompting (55%), and no longer initiating but willing to engage in the activity (42%; note that these groups can overlap). Many patients were also described as having less interest (44%) or impaired performance (34%) in the activity in which decreased initiation was noted. Coping strategies were employed by 69/85 carers, most commonly verbal prompts to begin an activity. CONCLUSION: Decreased initiation is a common and problematic symptom in people with mild-to-moderate Alzheimer's disease who seek treatment for dementia. Information offered voluntarily about interest in or ability to perform activities which are initiated less often gives additional data compared with routine questioning about initiation of function. This is a potentially rich area for the elucidation of disease and treatment effects. PMID- 18405400 TI - A supplementation study in human subjects with a combination of meso-zeaxanthin, (3R,3'R)-zeaxanthin and (3R,3'R,6'R)-lutein. AB - We measured the blood uptake of meso-zeaxanthin (MZ) from a mixture of macular pigments since its bioavailability in man has not been studied. Volunteers (ten men and nine women) were recruited and received one capsule of Lutein Plus/d. Blood was taken at baseline, day 10 and day 22. One capsule contained 10.8 mg lutein, 1.2 mg (3R,3'R)-zeaxanthin and 8.0 mg MZ. Plasma lutein and total zeaxanthin concentrations were quantified using isocratic liquid chromatography and the eluting xanthophyll fractions were collected and re-chromatographed on a chiral column to assess the proportion of MZ. Plasma concentrations per mg dose at day 22 suggested that (3R,3'R)-zeaxanthin (0.088 micromol/l per mg) was about 50 % more actively retained by the body than lutein (0.056 micromol/l per mg) (although the difference was not significant in women) and 2.5-3.0 times more than MZ (0.026 micromol/l per mg). Concentrations of MZ at day 22 were 2.5 times higher in women than men. The plasma responses from lutein and (3R,3'R) zeaxanthin in the Lutein Plus were lower than literature values for the pure substances. That is, their uptake into plasma appeared to be slightly depressed by the presence of MZ. Plasma concentrations of beta-carotene were depressed by about 50 % at day 10 and about 35 % at day 22. In conclusion, the lower plasma response to MZ compared with (3R,3'R)-zeaxanthin probably indicates that MZ is less well absorbed than (3R,3'R)-zeaxanthin but work with pure MZ will be needed to confirm that the lower plasma response was not due to the large amount of lutein in the Lutein Plus. PMID- 18405401 TI - The distributions and correlates of serum albumin levels in institutionalised individuals with intellectual and/or motor disabilities. AB - The serum albumin level is a widely accepted indicator of nutritional status in healthy adults and elderly individuals. However, there are few data regarding the distribution and correlates of serum albumin levels in individuals with intellectual and/or motor disabilities. We conducted a cross-sectional study at a public facility for individuals with intellectual and/or motor disabilities in Ibaraki prefecture, Japan. Health check-up data obtained in 2001 from 477 individuals with intellectual disability (286 males, average age 40.6 (SD 12.3) years; 191 females, average age 45.1 (SD 11.6) years) were retrospectively reviewed. With the exception of men with cerebral palsy, the serum albumin level was nearly normally distributed. The mean was 44 (SD 3.6) g/l for males and 44 (SD 3.6) g/l for females with intellectual disability, 43 (SD 3.2) g/l in males and 44 (SD 2.6) g/l for females with cerebral palsy, 41 (SD 2.7) g/l for males and 42 (SD 3.7) g/l for females with Down's syndrome, and 42 (SD 4.5) g/l for males and 41 (SD 3.2) g/l for females with severe motor and intellectual disabilities. Only six of 474 individuals (1.3 %) had a serum albumin 1 and d) gene expression of AMACR, HER-2/neu, high Bcl-2: Bax ratio and EZH2 in cancer cells. These markers have been chosen for review because they are among the most promising markers emerging currently. PMID- 18405442 TI - Treatment options for metastatic renal cell carcinoma: a review. AB - Metastatic renal cell carcinoma (RCC) has a poor overall survival. Localized RCC remains a surgical disease. About 20%-30% patients who present with limited disease at the time of nephrectomy develop metastasis. The median time to relapse after nephrectomy is 15-18 months. The maximum numbers of relapses are within the first 3 years. In metastatic RCC, immunotherapy is effective in a relatively small percentage of patients but is very toxic. In recent years, there has been an improved understanding of the biology of RCC. This has lead to the development of various agents that target ligands at the molecular level. The hypoxia inducible factor-alfa (HIF-)/ vascular endothelial growth factor (VEGF) pathway and mammalian target of rapamycin (mTOR) signal transduction pathway are targets for some of these novel agents. Recent randomized phase III trials have shown an improved outcome in patients with metastatic disease who received these targeted agents. This review deals with management of advanced and metastatic renal cell cancer with an emphasis on recently developed targeted therapies. PMID- 18405444 TI - Time to treatment of prostate cancer through the Calgary Prostate Institute rapid access clinic. AB - PURPOSE: To determine the wait time between initial referral, biopsy, diagnosis and individual treatment modalities of prostate cancer treatment through the Calgary Prostate Institute rapid access clinic (RAC) and compare to historical data estimates in Alberta and to suggested standards. Biopsy rate, rate of confirmed prostate cancer and the distribution of treatment modality for patients seen through the RAC is included. MATERIALS AND METHODS: A non-consented, retrospective chart review of 1103 patients from the Calgary Health Region referred to the RAC between September 2005 and August 2006 was completed. RESULTS: Patients experienced a median wait time of 21 days between referral from their family doctor and prostate biopsy. A total of 31.4% of patients referred to the clinic were requested to have a prostate biopsy performed and 50.8% of biopsies resulted in confirmed prostate cancer requiring treatment. Median wait time between diagnosis and treatment for all treatment types was 52.0 days with a 90th percentile of 146.2 days. Median wait time between referral and treatment for all treatment modalities was 101 days with a 90th percentile of 187.2 days. CONCLUSION: Calgary rapid access clinic reduces wait time between referral and biopsy by 78%. Stratifying across treatment type indicates that watchful waiting is the shortest time duration and radiation with hormone therapy is the longest. PMID- 18405445 TI - Radiofrequency ablation for T1a tumors in a solitary kidney: promising intermediate oncologic and renal function outcomes. AB - OBJECTIVE: The experience with radiofrequency ablation (RFA) in solitary kidneys is limited in numbers and follow-up. Therefore, we report our oncologic and renal function outcomes of RFA for T1a tumors in patients with a solitary kidney. METHODS: From April 2000 to August 2007, 242 patients were treated with RFA for renal cortical masses. Sixteen patients with localized tumors < or = 4 cm in a solitary kidney were identified. Clinical and radiographic data were reviewed to assess indications, complications, disease recurrence, and renal function. RESULTS: Twenty-one renal masses were ablated in 16 patients with a solitary kidney. The mean patient age was 66.1 years, and the mean tumor size was 2.6 cm (range, 1.1-4.0). Preoperative biopsy was diagnostic of renal cell carcinoma (RCC) in 75% of cases. At a mean follow-up of 30.7 months (range, 1.5-66.0), 14/16 (88%) patients had no radiographic evidence of disease recurrence. One patient with three masses in a solitary kidney had a local recurrence managed by salvage RFA. The other patient with a local failure also had severe baseline renal insufficiency with progression to end stage renal disease 2.5 years following RFA and underwent a radical nephrectomy. Mean glomerular filtration rate (GFR) decreased from 54.2 ml/min/1.73m2 preoperatively to 47.5 ml/min/1.73m2 at last follow-up (p = 0.015). There were no major complications, and four patients had minor complications which resolved without intervention. CONCLUSION: Radiofrequency ablation is an attractive alternative for the management of patients with T1a renal cortical tumors in a solitary kidney. Renal function appears to be adequately maintained with promising oncologic outcomes at 2.5 years. PMID- 18405446 TI - Routine postoperative chest radiography is not needed after flank incisions with eleventh rib resection. AB - INTRODUCTION: We wanted to determine whether routine postoperative chest radiography is needed after surgery with eleventh rib resection. MATERIALS AND METHODS: Data on 80 patients who underwent radical or partial nephrectomy, nephroureterectomy or adrenalectomy through 82 flank incisions with eleventh rib resection were collected and analyzed retrospectively. RESULTS: Radical and partial nephrectomies, nephroureterectomies and adrenalectomies were done through 47, 20, 6 and 9 flank incisions in 80 patients, respectively. Among these, one patient underwent a partial nephrectomy and subsequent contralateral radical nephrectomy, and another patient underwent simultaneous bilateral adrenalectomies. The intrapleural space was entered accidentally in 16 flank incisions (19.51%). Repair was performed with a simple evacuation technique at the time of surgery, and a chest tube was needed in only three patients (18.75%) according to postoperative chest x-ray evaluation after pleural repair. Of the remaining patients who had no evidence of pleural opening on the water seal test before wound closure, none developed respiratory distress postoperatively and no chest x-rays were ordered, except for one patient who developed subcutaneous emphysema on the day of surgery where no evidence of pneumothorax was detected. CONCLUSIONS: We concluded that when pleural injury was not observed and confirmed via the water seal test, none of the patients developed respiratory distress, and this suggests that there is no need to check for its presence with chest x-ray postoperatively. PMID- 18405447 TI - Erectile function following unilateral cavernosal nerve replacement. AB - INTRODUCTION: With nerve-sparing techniques, patients undergoing a radical prostatectomy may avoid the morbidity of erectile dysfunction. Certain patients who are not candidates for nerve-sparing procedures may be eligible for nerve interposition grafts. While bilateral cavernosal nerve grafting after radical prostatectomy has shown efficacy, the effect of unilateral nerve grafting following prostatectomy remains unclear. We evaluate a large group of patients who underwent a unilateral cavernosal nerve replacement. METHODS: Forty patients underwent unilateral nerve sparing surgery with concomitant contralateral cavernosal nerve replacement. Patients were selected for this procedure based upon preoperative nomogram risk assessment, endorectal MRI evidence of extra capsular disease (ECE) or intraoperative histology demonstrating margin positivity. Age, demographic data, Gleason score, clinical and pathologic stage and pre and post operative IIEF data was collected and prospectively analyzed. RESULTS: Median follow-up was 19 months. Median change in IIEF scores was 7.5. Twenty-one of 29 patients (72%) report being able to penetrate after prostatectomy. Sixteen of those 21 (76%) continue to require PDE-5 inhibitors to facilitate penetration. Four of the 6 patients (67%) who were unable to have intercourse following cavernosal nerve replacement received adjuvant hormonal and/or radiation therapy. Twenty-eight patients (97%) reported numbness at the graft harvest site. One patient experienced a graft site infection. Two of 29 (7%) patients reported pain at the harvest site. CONCLUSION: Unilateral sural nerve grafting is a feasible and well-tolerated approach for patients who must undergo wide resection of a NVB. While men do show a decrease in their IIEF score, 76% are able to achieve penetration following surgery. The majority of men continue to require PDE-5 inhibitors to facilitate intercourse. PMID- 18405448 TI - The addition of robotic surgery to an established laparoscopic radical prostatectomy program: effect on positive surgical margins. AB - PURPOSE: The addition of robotic assistance with the da Vinci surgical system for performing laparoscopic radical prostatectomy has been reported to improve surgical outcomes. In order to evaluate the benefit of robotic assistance to improve cancer control in a center with an established laparoscopic radical prostatectomy program, we evaluated the incidence of positive surgical margins in both transperitoneal laparoscopic (LRP) and robotically assisted laparoscopic radical prostatectomy (RALP). MATERIALS AND METHODS: We performed an Institutional Review Board (IRB) approved, retrospective review of 247 men with clinically localized prostate cancer treated with either a LRP or a RALP from March 2000 to August 2006. Pathology reports were reviewed for both preoperative and postoperative Gleason score as well as clinical and pathological stage. Surgical pathology specimens were evaluated using a whole mount, step section technique. Extracapsular extension, seminal vesicle invasion and positive margins were noted when present in the final surgical pathologic specimens. RESULTS: One hundred ninety seven patients underwent LRP, and 50 patients underwent RALP. Seven of the 197 LRP required open conversion to retropubic radical prostatectomy, and were excluded. None of the RALP were converted. The overall positive surgical margin rate for LRP and RALP was 18% (35/190) and 6% (3/50), respectively (p = 0.032). When examining pathologically organ confined specimens (pT2), the positive surgical margin rate was 12% (20/161) and 4.7% (2/43) for the LRP and RALP cohorts, respectively (p = 0.181). For pathologic disease that has spread outside the capsule (pT3/T4), the positive surgical margin rate was 54% (15/28) and 14% (1/7) for LRP and RALP, respectively (p = 0.062). Patient age, race and prostate volume were not significant factors in the incidence of positive surgical margins. CONCLUSION: The addition of robotic assistance to an established laparoscopic radical prostatectomy program appears to reduce the incidence of positive surgical margins. Data is maturing to determine whether this will lead to improved functional and oncologic outcomes. PMID- 18405449 TI - Hydrodissection for preservation of neurovascular bundle during robot-assisted radical prostatectomy. AB - INTRODUCTION: To describe a technique that may facilitate neurovascular bundle preservation during robot-assisted radical prostatectomy. MATERIALS AND METHODS: From December 2007 to January 2008, 10 patients underwent robot-assisted radical prostatectomy with bilateral nerve preservation. Hydrodissection of the neurovascular bundle was performed by injecting a 1:10000 epinephrine solution diluted in 0.9% normal saline into the lateral prostatic pedicle with an injection cannula needle (Wolf). Operative time, blood loss and margin status were assessed when this new technique was utilized. Erectile function status will be analyzed in the future. RESULTS: Ten potent patients underwent bilateral nerve sparing robot-assisted radical prostatectomy with hydrodissection. Mean patient age was 54 years old. Mean preoperative Gleason score was 6.5 and mean pretreatment PSA was 7.0. Six patients were clinical stage T1c and four patients were T2a. The mean operative time was 182 minutes, with a range of 148 minutes to 230 minutes. Mean blood loss was 297 cc. Hemodynamic changes were not seen during hydrodissection or after hydrodissection. No intraoperative or postoperative complications developed. None of the ten patients developed delayed postoperative bleeding. Final pathologic stage was pT2 in eight patients, pT3 in one patient and pT4 in one patient. All surgical margins were negative, except in the patient with bladder neck invasion. CONCLUSIONS: We describe an athermal technique which may facilitate neurovascular bundle preservation. While intraoperative parameters were favorable with hydrodissection, long term sexual function results need to be analyzed. PMID- 18405450 TI - The surgical technique and early postoperative complications of the Gynecare Prolift pelvic floor repair system. AB - INTRODUCTION: The Gynecare Prolift pelvic floor repair system (GPS) comprises a synthetic mesh placed via a transvaginal, transobturator approach. We present our technique focusing on the safety and feasibility of the GPS. MATERIALS AND METHODS: GPS candidates are evaluated in the office with a full history, physical examination, urinalysis and when appropriate, urodynamic evaluation. Patients were offered total vaginal vault prolapse repair or isolated anterior repair dependent of site of defect. Follow-up comprised a full history, physical examination, and global assessment of subjective satisfaction (2 and 6 weeks, 6 months postoperative). Concentration was placed on intraoperative and short-term postoperative complications and assessment of prolapse recurrence. RESULTS: GPS prolapse repair has been used in 12 patients for anterior or total vault prolapse. Mean postoperative follow-up time is 42 weeks. There were no major perioperative complications. De novo enterocele development was seen in one patient without any other incidence of recurrence. No incidence of mesh erosion or sexual dysfunction has been observed. CONCLUSIONS: The GPS is a safe and reproducible system for use in transvaginal repairs of vaginal vault prolapse. Long-term studies are needed to evaluate repair durability and for potential complications. PMID- 18405451 TI - A rare case of urinary bladder leiomyosarcoma accompanied by prostate cancer. AB - Non-urothelial neoplasms of the bladder account for less than 5% of all bladder tumors. Sarcoma constitutes the most usual mesenchymal malignancy of the bladder, with leiomyosarcomas being the most common type of sarcoma in adults. A coexistence of this rare tumor with another malignancy has not been reported, to our knowledge. This report demonstrates a case of high-grade bladder leiomyosarcoma cancer with prostate cancer in a 72-year-old patient. The most striking features of this case include the rapid disease progression that confirms the highly aggressive nature of this uncommon disease. PMID- 18405452 TI - Renal angiomyolipoma with intravascular extension into the inferior vena cava: a case report and review of the literature. AB - A 42-year-old woman with tuberous sclerosis (TS) presented with acute right-sided flank pain. Imaging studies demonstrated numerous bilateral renal angiomyolipomas (AML), the largest located within the right upper pole, extending into the venous system to the level of the infra-hepatic inferior vena cava (IVC). Intravascular extension of AML is quite rare, however, may potentially result in fatal complications if not appropriately treated. We present a case report and a description of the surgical management, and provide a review of the literature concerning this rare finding. PMID- 18405453 TI - Synchronous renal cell carcinoma and adrenocortical carcinoma: a rare case report and clinicopathologic approach. AB - A case of synchronous adrenocortical carcinoma (ACC) and renal cell carcinoma (RCC) has not yet been described in the English medical literature, to our knowledge. We report a first such case of adrenocortical and renal cell carcinomas occurring simultaneously in a 53-year-old male. He presented with history of vague abdominal pain. Ultrasound followed by a computed tomography (CT) scan and a magnetic resonance imaging (MRI) examination revealed a 6.4 cm left adrenal mass and a 3.5 cm right renal mass. The patient had complaints of gastroparesis manifesting with constant nausea as well as intermittent abdominal bloating and abdominal pain. He also had history of profuse intermittent sweating. There was no history of palpitations or fluctuations in blood pressure. The patient's urinary vanillylmandelic acid (VMA) levels and serum cortisol levels were normal. His 24-hour urine metanephrine levels were slightly elevated. Left adrenalectomy and right partial nephrectomy were performed. In this case, it is important to determine whether these tumors represent metastases or two synchronous tumors, as this has implications on the patient's management and prognosis. Clinical and pathological clues that led to the diagnosis are discussed in detail. PMID- 18405454 TI - Massive scrotal hematoma following transfemoral cardiac catheterization. AB - Scrotal hematoma is a rare and unusual complication of transfemoral cardiac catheterization. We describe the case of a 43-year-old man who presented with a delayed retroperitoneal hemorrhage and massive scrotal hematoma following cardiac catheterization. Emergent groin exploration was performed with ligation of the inferior epigastric artery and evacuation of the scrotal hematoma. Urological sequelae from transfemoral arterial access are rare. However, scrotal hematoma may occur if bleeding occurs between anatomic fascial planes. PMID- 18405455 TI - Bladder diverticulum arising adjacent to an ectopic ureter presenting as a cystic mass. AB - Acquired bladder diverticula due to bladder outlet obstruction are not uncommon in the adult male population. Congenital diverticula originate adjacent to the trigone and are rarely diagnosed in adults. We report an unusual case of a diverticulum arising adjacent to an ectopic ureter located on the left lateral wall near the dome of the bladder. Although the diverticulum appeared to be congenital, its large size was likely a result of high pressure voiding. The patient underwent a transurethral resection of the prostate to reduce his bladder outlet obstruction, and subsequently underwent an open diverticulectomy. PMID- 18405456 TI - Malignant melanoma of the prostate: a case report. AB - Primary genitourinary melanoma accounts for less than 1% of all cases of melanoma. Melanoma of prostatic origin is extremely rare. These patients are difficult to diagnose and carry a very poor prognosis. Aggressive surgical resection is the current treatment standard. We report a case of primary malignant melanoma of the prostate found during transurethral resection of the prostate. PMID- 18405457 TI - Office based non-oncology urology trials. PMID- 18405458 TI - Open clinical uro-oncology trials in Canada. PMID- 18405459 TI - Premature red blood cells have decreased aggregation and enhanced aggregability. AB - Preterm infants are highly susceptible to ischemic damage. This damage is most obvious in the brain, retina, and gastrointestinal tract. Studies focusing on the rheological properties of premature red blood cells (pRBCs) have consistently shown minimal or no RBC aggregation. Previously, measurements of pRBC aggregation kinetics indicated that specific plasma properties are responsible for the decreased RBC aggregation observed in the neonates, but that their specific RBC properties do not affect it. However, the strength of interaction in the pRBC aggregates as a function of medium composition has not been tested. In our previous research, we described clinically relevant parameters, that is, the aggregate resistance to disaggregation by flow. With the help of a cell flow property analyzer (CFA), we can monitor RBC aggregation by direct visualization of its dynamics during flow. We used the CFA to examine pRBC (from 9 premature babies) in the natural plasma and in PBS buffer supplemented with dextran (500 kDa) to distinguish between RBC intrinsic-cellular and plasma factors. pRBCs suspended in the native plasma showed minimal or no aggregation in comparison to normal adult RBC. When we transferred pRBCs from the same sample to the dextran solution, enhanced resistance to disaggregation by flow was apparent. PMID- 18405460 TI - So much done--so much to do. PMID- 18405461 TI - President's message. Recognizing the expertise in rural and remote medicine. PMID- 18405462 TI - Oligoanalgesia in a rural emergency department. AB - OBJECTIVE: Multiple studies conducted over many years have demonstrated that pain is poorly managed in the emergency department (ED). This phenomenon has been referred to in the medical literature as "oligoanalgesia." However, little is known about whether oligoanalgesia occurs in a rural ED. National Ambulatory Care Reporting System data from 2003 for a small rural hospital in Ontario showed patients were satisfied with the amount of pain medicine they received in the ED. We designed a study to replicate a published urban study that investigated the use of analgesia in isolated lower limb injuries. Our objective was to see if oligoanalgesia was also a problem in a rural ED. METHODS: In 2003 we conducted a retrospective chart review of patients who presented to the South Huron Hospital ED with isolated lower extremity injuries for which radiographs of the foot, ankle or both were obtained. Demographics of the ED patients with lower extremity injuries were quantified. Other parametres included whether or not patients received analgesia in the ED; how long it took to get assessed, treated and discharged; whether patients received any analgesia upon discharge; what type of analgesia they received; and whether it required a prescription. RESULTS: A total of 189 patients met inclusion criteria, with 35 fractures identified (18.5%). Sixty-three percent of patients were male. The average age was 32.6 years. The mean Canadian Emergency Department Triage and Acuity Scale level was 4.4. The mean time to physician assessment was 31.6 minutes. The mean length of time spent in the ED was 74 minutes. Over one-half of the patients received analgesia upon discharge from the ED whether or not they had a fracture. In addition, 73% of the people in the fracture group received analgesia requiring a prescription, versus only 46% in the nonfracture group. Narcotics were used more often in the fracture group than in the nonfracture group (26% v. 6%). CONCLUSION: The phenomenon of oligoanalgesia was not observed as often in our rural ED for isolated lower limb injuries, when compared with the published urban study. PMID- 18405463 TI - An analysis of appendectomies performed in a Labrador general surgery practice. AB - INTRODUCTION: The main objective of our study was to determine the rates of negative appendectomies and perforated appendices at the Labrador Health Centre, and make a comparison with the rates published in the literature. METHODS: The study population consisted of all patients who underwent appendectomies during the 5-year period ending Apr. 3, 2006. The number and rates of negative appendectomies and perforated appendices were determined for each age and sex category. RESULTS: Of the 64 patients who were included in the study, 11% were found to have undergone negative appendectomies and 27% had perforated appendices. There was a clear trend toward decreasing perforation rates with increasing age as well as a trend toward increasing negative appendectomy rates with increasing age. CONCLUSION: The rates of negative appendectomies and perforated appendices at the Labrador Health Centre are comparable with those published in the literature. Trends found in the data will help to guide future improvements in patient management. PMID- 18405464 TI - The difference between medical students interested in rural family medicine versus urban family or specialty medicine. AB - OBJECTIVE: To determine how first-year medical students interested in rural family medicine in Canada differ from their peers. METHOD: From 2002 to 2004, first-year students (n = 2189) from 16 classes in 8 Canadian medical schools ranked intended career choices and indicated influences on their choices using Likert scales. We used t tests and chi2 tests to determine demographic influences and factor analysis, and we used analysis of variance to examine associated attitudes. RESULTS: Of the 1978 surveys returned (90.3%), 1905 were used in the analysis. Rural family medicine was ranked first by 11.1%, varying from 4.7% to 20.2% among schools. Students interested in rural family medicine were more likely to have grown up rurally, graduated from a rural high school and have family in a rural location than others (p < 0.001). They were more likely to be older, in a relationship, to have volunteered in a developing nation and less likely to have university-educated parents than those interested in a specialty (p < 0.008). Attitudes of students choosing family medicine, rural or urban, include social orientation, preference for a varied scope of practice and less of a hospital orientation or interest in prestige, compared with students interested in specialties (p < 0.001). CONCLUSION: Medical schools may address the rural physician shortages by considering student demographic factors and attitudes at admission. PMID- 18405465 TI - The occasional intraosseous infusion. PMID- 18405466 TI - Very rural obstetrics. PMID- 18405467 TI - Researching rural: research interns in northern Ontario. PMID- 18405468 TI - Payment to healthcare professionals for patient recruitment to trials: systematic review and qualitative study. AB - OBJECTIVES: To review UK guidelines regarding the use of financial incentives for healthcare professionals to become involved in clinical trials, and to survey perceptions and current practice. DATA SOURCES: Electronic databases were searched from inception to June 2006. Interviews were held with NHS healthcare professionals, research managers from the pharmaceutical industry and members of the public. REVIEW METHODS: From the searches, 634 identified studies were assessed for inclusion in the systematic review, but only three met the criteria for data extraction. Fifty-eight individuals were interviewed: 38 chief investigators, six non-research active clinicians, eight public and six pharmaceutical managers. Investigators were selected from those funded by the HTA Programme, the other by 'snowballing' and personal contact. RESULTS: The evidence from the literature was limited and inconclusive. In UK guidelines, the issues around payments to clinicians or patients were implied rather than stated, usually linked to discussion of conflict of interest and disclosure of any such conflicts. Developments in NHS research governance had led to increased transparency in all payments for research participation and for payments to be made to NHS Trusts rather than individual clinicians. While reimbursement of costs incurred by research was strongly supported by the interviewees, payments to incentivise recruitment were not. A code of practice was suggested for payments in publicly funded trials, which was closely linked to the principles of Good Clinical Practice in research. Factors such as interest in the topic, scope for patient benefit and good communication were considered more important than payment. Interviews with the general public indicated low levels of awareness of the existence of payments to clinicians linked to patient recruitment in trials, and unanimous support for full disclosure. Interviews with managers in the pharmaceutical industry showed greater familiarity with payments for research involvement. GPs were seen as the only group for whom scope existed for individual payments. Concerns were expressed by the pharmaceutical company interviewees at the rising cost of research and unnecessary bureaucracy. CONCLUSIONS: The ethical stances outlined in Good Clinical Practice in research were widely endorsed by the three groups interviewed. These allow reasonable payments to clinicians, subject to disclosure of any possible conflicts of interest. The potential for incentivising clinicians to recruit was limited as any payments should be based on the cost of inputs and should not be made to individuals but to their host organisation. NHS professionals were concerned that payments could damage the quality of research and also considered full disclosure to patients as challenging. Patients and members of the public favoured full disclosure and payment of expenses to patients involved in research. Pharmaceutical company interviewees viewed payment to the NHS for all research activities as normal and highly regulated. They complained that the prices charged were high and so variable that they required benchmarking. Considerable scope exists for compiling data on the factors that help and hinder the progress of clinical trials and also for experimenting with different incentives to encourage involvement in clinical research. Further research should focus on improved reporting of those organisational aspects of trials that are known to affect recruitment; retrospective analysis of the factors associated with different levels of recruitment to RCTs; prospective comparative research on trial recruitment; qualitative research on participants' experiences of being involved in different kinds of trials, and proposals to include within trials experiments with payments methods. PMID- 18405469 TI - The clinical effectiveness of diabetes education models for Type 2 diabetes: a systematic review. AB - OBJECTIVE: To examine the clinical effectiveness of patient education models for adults with Type 2 diabetes. DATA SOURCES: Electronic databases were searched from 2002 to January 2007. REVIEW METHODS: A systematic review of the literature on educational interventions in diabetes was undertaken. This was an update of a previous systematic review. RESULTS: Including studies identified in the previous systematic review, there were 13 published studies. Eight studies of education on multiple aspects of diabetes self-management were identified that provided education that was focused on a particular aspect of self-management. The quality of reporting and methodology of the studies was variable. Studies of multi component educational interventions yielded mixed results. Some trials reported significant improvements on measures of diabetic control but others did not. Positive effects may be attributable to longer-term interventions with a shorter duration between the end of the intervention and the follow-up evaluation point. There may also be an effect of having a multi-professional team delivering the educational programme. Studies of focused educational interventions did not yield consistent results. Some effects were shown on measures of diabetic control in studies that focused on diet or exercise alone. Although the effects shown were generally small, those that were present did appear to be relatively long lasting. This update review does not substantially alter the conclusions of the previous systematic review; for each outcome, the proportion of studies that demonstrated significant effects of education was similar. CONCLUSIONS: Based on the evidence, it would seem that education delivered by a team of educators, with some degree of reinforcement of that education made at additional points of contact, may provide the best opportunity for improvements in patient outcomes. Educators need to have time and resources to fulfil the needs of any structured educational programme. There is also a need for education to have a clear programme at the outset. From the evidence reported it is unclear what resources would need to be directed at the educators themselves to ensure that they can deliver programmes successfully. Any future research should consider patient education within the context of overall diabetes care and as such follow guidelines for the development and evaluation of complex interventions. Good quality, longer-term studies would be desirable, but these would require careful consideration around the nature of any control group. Information is needed to clarify the sensitivity of diabetes education programmes to the performance of the diabetes educators, in order to ensure success and cost-effectiveness of education programmes. PMID- 18405470 TI - Cyclooxygenase-2 selective non-steroidal anti-inflammatory drugs (etodolac, meloxicam, celecoxib, rofecoxib, etoricoxib, valdecoxib and lumiracoxib) for osteoarthritis and rheumatoid arthritis: a systematic review and economic evaluation. AB - OBJECTIVES: To review the clinical effectiveness and cost-effectiveness of cyclooxygenase-2 (COX-2) selective non-steroidal anti-inflammatory drugs (NSAIDs) (etodolac, meloxicam, celecoxib, rofecoxib, etoricoxib, valdecoxib and lumiracoxib) for osteoarthritis (OA) and rheumatoid arthritis (RA). DATA SOURCES: Electronic databases were searched up to November 2003. Industry submissions to the National Institute for Health and Clinical Excellence (NICE) in 2003 were also reviewed. REVIEW METHODS: Systematic reviews of randomised controlled trials (RCTs) and a model-based economic evaluation were undertaken. Meta-analyses were undertaken for each COX-2 selective NSAID compared with placebo and non-selective NSAIDs. The model was designed to run in two forms: the 'full Assessment Group Model (AGM)', which includes an initial drug switching cycle, and the 'simpler AGM', where there is no initial cycle and no opportunity for the patient to switch NSAID. RESULTS: Compared with non-selective NSAIDs, the COX-2 selective NSAIDs were found to be equally as efficacious as the non-selective NSAIDs (although meloxicam was found to be of inferior or equivalent efficacy) and also to be associated with significantly fewer clinical upper gastrointestinal (UGI) events (although relatively small numbers of clinical gastrointestinal (GI) and myocardial infarction (MI) events were reported across trials). Subgroup analyses of clinical and complicated UGI events and MI events in relation to aspirin use, steroid use, prior GI history and Helicobacter pylori status were based on relatively small numbers and were inconclusive. In the RCTs that included direct COX-2 comparisons, the drugs were equally tolerated and of equal efficacy. Trials were of insufficient size and duration to allow comparison of risk of clinical UGI events, complicated UGI events and MIs. One RCT compared COX-2 (celecoxib) with a non-selective NSAID combined with a gastroprotective agent (diclofenac combined with omeprazole); this included arthritis patients who had recently suffered a GI haemorrhage. Although no significant difference in clinical GI events was reported, the number of events was small and more such studies, where patients genuinely need NSAIDs, are required to confirm these data. A second trial showed that rofecoxib was associated with fewer diarrhoea events than a combination of diclofenac and misoprostol (Arthrotec). Previously published cost effectiveness analyses indicated a wide of range of possible incremental cost per quality-adjusted life-year (QALY) gained estimates. Using the simpler AGM, with ibuprofen or diclofenac alone as the comparator, all of the COX-2 products are associated with higher costs (i.e. positive incremental costs) and small increases in effectiveness (i.e. positive incremental effectiveness), measured in terms of QALYs. The magnitude of the incremental costs and the incremental effects, and therefore the incremental cost-effectiveness ratios, vary considerably across all COX-2 selective NSAIDs. The base-case incremental cost per QALY results for COX-2 selective NSAIDs compared with diclofenac for the simpler model are: celecoxib (low dose) 68,400 pounds; celecoxib (high dose) 151,000 pounds; etodolac (branded) 42,400 pounds; etodolac (generic) 17,700 pounds; etoricoxib 31,300 pounds; lumiracoxib 70,400 pounds; meloxicam (low dose) 10,300 pounds; meloxicam (high dose) 17,800 pounds; rofecoxib 97,400 pounds; and valdecoxib 35,500 pounds. When the simpler AGM was run using ibuprofen or diclofenac combined with proton pump inhibitor (PPI) as the comparator, the results change substantially, with the COX-2 selective NSAIDs looking generally unattractive from a cost-effectiveness point of view (COX-2 selective NSAIDs were dominated by ibuprofen or diclofenac combined with PPI in most cases). This applies both to 'standard' and 'high-risk' arthritis patients defined in terms of previous GI ulcers. The full AGM produced results broadly in line with the simpler model. CONCLUSIONS: The COX-2 selective NSAIDs examined were found to be similar to non-selective NSAIDs for the symptomatic relief of RA and OA and to provide superior GI tolerability (the majority of evidence is in patients with OA). Although COX-2 selective NSAIDs offer protection against serious GI events, the amount of evidence for this protective effect varied considerably across individual drugs. The volume of trial evidence with regard to cardiovascular safety also varied substantially between COX-2 selective NSAIDs. Increased risk of MI compared to non-selective NSAIDs was observed among those drugs with greater volume of evidence in terms of exposure in patient-years. Economic modelling shows a wide range of possible costs per QALY gained in patients with OA and RA. Costs per QALY also varied if individual drugs were used in 'standard' or 'high'-risk patients, the choice of non-selective NSAID comparator and whether that NSAID was combined with a PPI. With reduced costs of PPIs, future primary research needs to compare the effectiveness and cost-effectiveness of COX-2 selective NSAIDs relative to non-selective NSAIDs with a PPI. Direct comparisons of different COX-2 selective NSAIDs, using equivalent doses, that compare GI and MI risk are needed. Pragmatic studies that include a wider range of people, including the older age groups with a greater burden of arthritis, are also necessary to inform clinical practice. PMID- 18405472 TI - [Can deep venous thrombosis be excluded by D-dimer test?]. PMID- 18405471 TI - The clinical effectiveness and cost-effectiveness of central venous catheters treated with anti-infective agents in preventing bloodstream infections: a systematic review and economic evaluation. AB - OBJECTIVES: To assess the clinical effectiveness and cost-effectiveness of central venous catheters (CVCs) treated with anti-infective agents in preventing catheter-related bloodstream infection (CRBSI). DATA SOURCES: Major electronic databases were searched from 1985 to August 2005. REVIEW METHODS: The systematic clinical and economic reviews were conducted according to accepted procedures. Only full economic evaluations (synthesis of costs and benefits) comparing the use of anti-infective central venous catheters (AI-CVCs) with untreated CVCs or other treated catheters were selected for inclusion in the economic review. RESULTS: A total of 32 trials met the clinical inclusion criteria. Seven different types of AI-CVC were identified, with the most frequently tested being chlorhexidine and silver sulfadiazine (CHSS) (externally treated), CHSS (externally and internally treated) and minocycline rifampicin (internally and externally treated). In general, the trials were of a poor quality in terms of reported methodology, microbiological relevance and control of confounding variables. The pooled result suggests a statistically significant advantage for AI-CVCs in comparison to standard catheters in reducing CRBSI [odds ratio (OR) 0.45, 95% confidence interval (CI) 0.34 to 0.60, 24 studies, I-squared = 0%, fixed effects]. Analysis by subgroups of catheters demonstrates that antibiotic treated catheters and catheters treated internally and externally decrease CRBSI rates significantly (OR 0.26, 95% CI 0.15 to 0.46, six studies, I-squared = 0%, fixed effects, and OR 0.43, 95% CI 0.26 to 0.70, nine studies, I-squared = 0%, fixed effects, respectively). Catheters treated only externally demonstrate a wider CI and non-significant effect (OR 0.67, 95% CI 0.43 to 1.06, nine studies, I-squared = 0%, fixed effects). A treatment effect was also found for trials with an average duration of between 5 and 12 days, and for the one study with a mean duration of over 20 days. There was a statistically significant treatment effect for both femoral and jugular insertion sites and for those studies reporting a mix of insertion sites. The treatment effect was not observed in trials using exclusively subclavian insertion sites. Of the four trials that compared treated catheters, one reported a benefit of antibiotic-treated catheters over catheters treated externally with CHSS. All three sensitivity analyses testing for study design differences reported a statistically significant treatment effect. The review was limited owing to the quality of the trials included, marked differences in the definitions and methods of diagnosis of CRBSI, and inconsistent reporting of risk factors and patient population factors. Furthermore, two-thirds of trials were commercially funded. The economic performance (cost-effectiveness and potential cost-savings) of using AI-CVCs to reduce the number of CRBSIs in patients requiring a CVC was also reviewed. Results show that the use of AI-CVCs instead of standard CVCs can lead to a reduction in CRBSIs and decreased medical costs. To complement the reviews, a basic decision-analytic model was constructed to explore a range of possible scenarios for the NHS in England and Wales. Results show that for every patient who receives an AI-CVC there is an estimated cost-saving of 138.20 pounds. The multivariate sensitivity analyses estimate potentially large cost-savings, depending on the size of the population, under a wide range of cost and clinical assumptions. However, those considering the purchase of AI-CVCs should ensure that their patient populations and the important characteristics of local clinical practice are indeed similar to those described in this economic evaluation. CONCLUSIONS: Overall, AI-CVCs are clinically effective and relatively inexpensive and therefore their integration into clinical practice can be justified. However, the use of these anti-infective catheters without the appropriate use of other practical care initiatives will have only a limited success on the prevention of CRBSIs. Comparative trials are required to determine which, if any, of the treated catheters is the most effective. Pragmatic research related to the effectiveness of bundles of care that may reduce rates of CRBSI is also warranted. PMID- 18405473 TI - [Diagnostic strategy in patients with clinically suspected deep vein thrombosis]. AB - INTRODUCTION: The standard method for diagnosing deep vein thrombosis (DVT) involves determination of D-dimer and ultrasound scanning. In an attempt to reduce the number of ultrasound examinations we have supplemented this with a clinical probability estimate for DVT (DVT-score) over one year. MATERIALS AND METHODS: A total of 508 consecutive patients presenting in the emergency room with suspected DVT had D-dimer and DVT-score performed. Patients with non elevated D-dimer and a low or moderate DVT score received no treatment. The remainder had ultrasound scanning from the groin to the popliteal fossa. If no DVT was revealed, the patient was contacted by telephone 7-10 days later, and was offered a repeat examination if symptoms persisted. RESULTS: Three patients with chronic DVT were excluded. Normal D-dimer and low or moderate DVT-score was found in 103 patients, none had DVT. Only five patients with normal D-dimer had high DVT-scores, none had DVT, so that the benefit from determining DVT-scores was modest. Ultrasound scanning revealed DVT in 85 out of 397 patients with elevated D-dimer. A repeat examination was performed in 91 patients with persisting symptoms, and disclosed DVT in two. CONCLUSION: We recommend that ambulatory patients with clinically suspected DVT have a D-dimer test. If D-dimer is elevated, compression ultrasound should be performed in the groin and the popliteal fossa. PMID- 18405474 TI - [Treatment of elderly patients with breast cancer]. AB - The latest investigations have been searched in order to present new guidelines for the treatment of elderly patients with primary breast cancer. It is concluded that breast-conserving surgery should be offered as well as the sentinel node technique. Axillary lymph node dissection is not necessary in the case of micrometastases. Adjuvant radiotherapy can be omitted in low risk cases. Endocrine treatment can be used in adjuvant and neo-adjuvant settings, but surgery should be offered as well. In high risk cases with receptor-negative tumour chemotherapy should be considered as an option. Generalized follow-up is recommended. PMID- 18405475 TI - [Multidisciplinary rehabilitation of patients with multiple sclerosis. A survey of a Cochrane review]. PMID- 18405476 TI - [Diving sickness]. PMID- 18405477 TI - [When medical records are the best witness]. AB - This article looks at the development in the increasing use of medical records in the Danish Courts as well as outside the courts in cases of personal injury. The Danish Supreme Court puts the presence of all material above the protection of the confidential relationship between doctor and patient. It is not yet clear to what extent the use of medical records will be accepted. This development raises questions regarding legal security for patients and sets higher requirements for medical and legal personnel. Medical records give important testimony in cases regarding personal injury. It is therefore important for medical personnel to be aware of the content of the medical record, as it might be used and interpreted in the courtroom in a different manner than intended. PMID- 18405478 TI - [How is out-of-hospital cardiac arrest dispatched?]. AB - INTRODUCTION: In Denmark any person needing urgent medical help can dial 112 and get in contact with an alarm centre where a non-health educated operator assesses what kind of help is needed. A specific dispatch report (DR) is used if an ambulance is dispatched. We assessed which DRs were used for the Copenhagen Mobile Emergency Care Unit (MECU) in the case of out-of-hospital cardiac arrest. MATERIALS AND METHOD: All DRs for the MECU during 2000 to 2006 were analyzed and compared with the diagnosis recorded by the dispatched specialist in anaesthesiology after every case. We divided the DRs into five categories: ''cardiac arrest'', ''possible death'', ''unconscious'', ''heart attack'', and ''miscellaneous'' (consisting of 40 different DR categories). RESULTS: We found 52088 DRs, 2902 of which were diagnosed as cardiac arrest. 32% of these cardiac arrests were dispatched in accordance with this, while the DRs were different from cardiac arrest in 68%. ''Unconscious'' accounted for 21%. 41% of the cases with DR cardiac arrest could not be verified upon the arrival of the dispatched medical doctor. CONCLUSION: Only 32% of the cases with cardiac arrest had a correct DR. We suspect that some of the patients had an unrecognized cardiac arrest at the time of contact to the alarm centre. The current alarm system can presumably be improved. The alarm centre has a central role in such a quality improvement. PMID- 18405479 TI - [Antiplatelet treatment after acute and chronic ischemic heart and brain events is insufficient according to recommendations]. AB - INTRODUCTION: Antiplatelet therapy is important in secondary prophylaxis in patients with ischemic heart, brain and vascular diseases. The aim of this study was to investigate the number of patients in Vejle County admitted to hospital in the second half of 2003 with acute and chronic heart, brain and vascular diseases to evaluate the kind of secondary prophylaxis and its duration. Moreover, the study registered the number of patients with diabetes mellitus treated with antiplatelet therapy. MATERIALS AND METHODS: The study included 2345 patients with ischemic heart, brain and vascular diseases, and patients with diabetes mellitus. Patients' data and information about medication were obtained from the hospital electronic database and public health insurance of Vejle County. Antiplatelet therapy was registered up to one year after onset of the disease. RESULTS: 1121 patients were admitted with cardiac diseases (including patients with angina pectoris), 624 patients had cerebral diseases, and 600 had diabetes mellitus. Acetylsalicylic acid (ASA) and a combination of ASA and clopidogrel were the most frequent forms of antiplatelet medication. Patients with diabetes and ischemic brain diseases made up 75% of those for whom medication was not registered. CONCLUSION: The study shows that in nearly all disease groups no antiplatelet therapy was registered for a very large number of % patients (up to 75%). A considerable number % of patients (up to 56%) with ischemic heart disease did not receive sufficient antiplatelet treatment according to Danish recommendations. PMID- 18405480 TI - [Prenatal diagnosis of chromosome aberrations after implementation of screening for Down's syndrome]. AB - INTRODUCTION: First trimester screening for Down's syndrome was evaluated by the National Board of Health in 2004, and recommended to all pregnant women in the form of an informed choice. We have reviewed prenatal and postnatal chromosome aberrations in 3 counties in Denmark during the years of implementation in 2004, 2005 and 2006. MATERIALS AND METHODS: Risk evaluation based on combined screening (fetal nuchal translucency measurement and serum screening of the pregnant woman) was introduced in the counties of Copenhagen, Roskilde and Storstrom, covering approximately 1.1 million inhabitants. We registered the number of chorionic villus biopsies (CVS) and amniocenteses (AC), as well as the number of cases with trisomy, triploidy and sex chromosome aberrations found prenatally. We also registered the number of children born with Down's syndrome during the period. RESULTS: The number of CVS/AC decreased from 1382 to 790, or 40%. There was an increase in the number of foetuses diagnosed with trisomy 21: in 2004 trisomy 21 was diagnosed in 12 foetuses, in 2006 the number was 30. The number of children born with Down's syndrome was 10 and 5 in 2004 and 2006, respectively. National figures from the Danish central cytogenetic registry confirm a decrease in children born with Down's syndrome. CONCLUSION: The implementation of combined screening in 3 counties resulted in a reduction in invasive procedures (chorionic villus samples and amniocenteses) by 40%, which is in accordance with the aims of the National Board of Health. As expected, a significant increase in the number of prenatally diagnosed foetuses with trisomy 21 was observed. The number of children born with Down's syndrome decreased, but the numbers are small. The investigation does not review aspects of organisation or counselling and psychosocial issues. PMID- 18405481 TI - [Coronary subclavian steal syndrome: two cases after coronary artery bypass grafting]. AB - Reverse flow in the internal mammary artery (IMA) graft due to stenosis or occlusion of the proximal ipsilateral subclavian artery causes coronary subclavian steal syndrome (CSSS). We describe two patients who were diagnosed with CSSS following CABG. Patient A presented with angina pectoris, was diagnosed with CSSS and treated by transposition of the proximal IMA from the subclavian artery to the aorta. Patient B was diagnosed with CSSS by control angiography. Myocardial scintigraphy showed reversible silent ischemia. He was offered treatment, but refused as he was symptom-free. PMID- 18405482 TI - [Actinic prurigo]. AB - A 7-year-old girl of Colombian descent was referred for diagnosis and treatment. She initially presented signs of universal itchy eczema in the spring of 2003. She responded to local treatment with topical steroids, but her symptoms rapidly re-emerged after discharge. Further investigations led to the conclusion that the patient suffered from Actinic Prurigo. The disease was very severe and there was a lack of response to treatment with oral prednisolone, antihistamines and pentoxyfyllin. Therefore the patient was treated with Thalidomide 50 mg/25 mg daily (alternating). This resulted in the complete clearance of her skin symptoms except postinflammatory hyperpigmentation. PMID- 18405483 TI - [Severe anorexia nervosa: five fatal cases]. AB - The Centre for Eating Disorders at Odense University Hospital receives about 140 patients yearly. We reviewed the five lethal cases that occurred during the last 10 years. None of the patients died from suicide, which is unexpected, because patients with anorexia nervosa (AN) are known to have an increased suicide rate. Although compulsory treatment has shown to be efficient in treating AN, only three of the five patients were compulsorily admitted and only one was admitted several times. Dropout remains a significant problem in the treatment of AN, as four out of the five patients died after they had dropped out of the treatment program. PMID- 18405484 TI - [Utility of endoscopic ultrasonography in the etiological diagnosis of patients with acute idiopathic pancreatitis]. AB - The aim of this study was to evaluate the utility of endoscopic ultrasonography in the etiological diagnosis of patients initially diagnosed with acute idiopathic pancreatitis and to determine the clinical and laboratory factors related to the results of this technique. MATERIAL AND METHODS: We studied 73 patients initially diagnosed with acute idiopathic pancreatitis and referred to our service for biliopancreatic endoscopic ultrasonography. A positive result was considered to be identification of at least one possible cause of pancreatitis, excluding patients in whom the only echoendoscopic finding was chronic pancreatitis. The clinical characteristics and clinical course of these patients were compared with the results of the technique. RESULTS: Endoscopic ultrasonography diagnosed at least one possible cause of acute pancreatitis in 32 patients (44%). The most frequent diagnoses were cholelithiasis (16%) and biliary sludge (20%). The diagnostic yield of endoscopic ultrasonography was lower in patients with prior cholecystectomy (16% vs. 49%; p=0.037) while a positive result was more likely in patients with elevated glutamyl oxaloacetic transaminase (GOT) or glutamyl pyruvic transaminase (GPT) levels at diagnosis of acute pancreatitis (GOT: 68 vs. 31%; p=0.002; GPT: 63 vs. 26%; p=0.001). No differences were found in diagnostic yield according to whether endoscopic ultrasonography was performed during the first episode of pancreatitis (48%) or in recurrent episodes (37%) (p=0.40). Over a mean follow-up period of 28 months, recurrence of new episodes of pancreatitis was significantly lower in patients with an etiological diagnosis (3 vs. 22%; p=0.02). CONCLUSIONS: Endoscopic ultrasonography is a useful technique in the etiological diagnosis of acute pancreatitis of unknown origin, especially in patients with elevated transaminase levels and/or without cholecystectomy. This technique should be used in the first episode of acute idiopathic pancreatitis. PMID- 18405485 TI - [Effect of Helicobacter pylori eradication on iron deficiency anemia of unknown origin]. AB - Iron deficiency anemia of unknown origin is a frequent cause of anemia in which etiological diagnosis is often not achieved, despite currently available diagnostic techniques. Recent studies suggest that, in the absence of digestive tract lesions, Helicobacter pylori infection could be the cause of iron deficiency anemia, due to the alterations produced in gastric iron absorption. OBJECTIVES: To evaluate whether H. pylori eradication resolves iron deficiency anemia and removes the need for oral iron administration. PATIENTS AND METHODS: We performed an observational descriptive study in patients with iron deficiency anemia refractory to treatment with oral iron administration and with out causes that could explain their anemia. Gastroscopy, ileocolonoscopy, intestinal transit study and/or endoscopic capsule were performed. Female patients also underwent gynecological study. All patients were H. pylori-positive and standard eradication therapy was administered until elimination was achieved. The patients were followed-up for a minimum of 3 months after H. pylori eradication and the need for oral iron intake after eradication was evaluated. RESULTS: Ten patients, aged 53+/-8.2 years, were included. Hemoglobin (Hbg) before treatment was 10.06+/ 0.53 mg/dl, mean corpuscular volume (MCV) was 75.43+/-6.02 fl and ferritin was 6.1+/-3.28 ng/ml. Eradication therapy was administered until elimination of H. pylori. The mean time before disappearance of anemia was 4.5 months. Laboratory parameters after treatment were as follows: Hgb 12.86+/-0.75 mg/dl, MCV 85.02+/ 4.8 fl and ferritin 28+/-22.19 ng/dl. CONCLUSIONS: In the absence of lesions that could explain iron deficiency anemia, this disease can be related to H. pylori infection. Eradication of this infection is closely followed by disappearance of anemia and ferropenia. PMID- 18405486 TI - [Gastric plasmacytoma as an uncommon cause of upper gastrointestinal bleeding]. AB - Gastric plasmacytoma is an infrequent form of presentation of monoclonal gammopathy that, if ulcerated, can cause upper gastrointestinal bleeding. This entity is usually localized but may progress to multiple myeloma or other forms of monoclonal gammopathy. Differential diagnosis should be performed with other monoclonal gammopathies, based on laboratory and histological criteria, as well as with other gastric tumors such as adenocarcinoma, gastrointestinal stromal tumors, neuroendocrine tumors and lymphomas. Differential diagnosis can be based on radiological and endoscopic characteristics, although definitive diagnosis will undoubtedly be based on histological study and immunohistochemical techniques. We describe the case of a male patient who underwent investigations for upper gastrointestinal bleeding and iron deficiency anemia. The final diagnosis was multiple myeloma associated with gastric plasmacytoma. PMID- 18405487 TI - [Intestinal lymphoma and mesenteric panniculitis: complications of undiagnosed celiac disease]. AB - Celiac disease is the most common severe food intolerance in the Western world and is due to gluten ingestion in genetically susceptible children and adults. The key treatment in these patients is a gluten-free diet, because most complications are more common when dietary compliance is poor. The most serious complication of celiac disease is the development of neoplasms (the most common of which is enteropathy-associated T-cell lymphoma). However, a number of reports have indicated an increased prevalence of ulcerative jejunitis and extraintestinal manifestations, including chronic hepatitis, fibrosing lung disease, and epilepsy syndromes. We report the case of a 53-year-old-man with long-standing diarrhea; because celiac disease was not suspected, the patient developed celiac-associated T-cell lymphoma and mesenteric panniculitis. PMID- 18405488 TI - [Acute intermittent porphyria and chronic transaminase elevation]. AB - Acute intermittent porphyria is an autosomal dominant inherited disorder resulting from a deficiency of porphobilinogen deaminase activity, the third enzyme in the heme biosynthesis pathway. This disease is uncommon, although the prevalence is higher in asymptomatic heterozygotic carriers; however, this prevalence is difficult to establish because of the absence of symptoms. Although acute intermittent porphyria is a multisystemic disease, its most common form of presentation is abdominal pain and neurological or mental symptoms, which can sometimes be due to precipitating factors such as reduced energy intake, smoking, alcohol, some drugs, and stress. Diagnosis can be made by testing urinary porphobilinogen levels, with subsequent measurement of enzyme activity and DNA testing. Treatment is based on prevention of porphyria attacks by avoiding precipitating factors and early administration of intravenous glucose or hemin therapy. We present the case of a patient diagnosed with acute intermittent porphyria based on study of chronic mild alanine aminotransferase (ALT) elevation. PMID- 18405489 TI - [Treatment of fatty liver disease]. AB - Nonalcoholic fatty liver disease (NAFLD) is considered to be a hepatic manifestation of metabolic syndrome. The clinicopathologic spectrum ranges from simple steatosis to nonalcoholic steatohepatitis (NASH). Simple steatosis has a relatively benign clinical course, but NASH can progress to cirrhosis and hepatocellular carcinoma. As yet there is no convincingly effective treatment for NAFLD and the best option for these patients might be a multimodal treatment plan targeting obesity, insulin resistance, diabetes mellitus, hyperlipidemia and hypertension. PMID- 18405490 TI - [How can we find, design, evaluate and use clinical practice guidelines?]. PMID- 18405491 TI - [Telbivudine in the treatment of the chronic B hepatitis]. PMID- 18405492 TI - [Pancreatitis associated with metronidazole]. PMID- 18405493 TI - [Rectorrhagia due to oxygenated water]. PMID- 18405494 TI - [Commentaries on the article "are there frequent errors in the management of outpatients with inflammatory bowel disease?"]. PMID- 18405495 TI - [Subcapsular hepatic hematoma secondary to the use of hydrophilic guidewires during endoscopic retrograde cholangiopancreatography]. PMID- 18405497 TI - [Study of aerobic capacity in chronic hemodialized patients: effect of L carnitine supplementation]. AB - BACKGROUND AND OBJECTIVE: Chronic hemodialyzed patients have a low level of aerobic capacity, caused by the pathologies concomitant to renal insufficiency, according with a low level of physical activity. One of the factors that would contribute to this level of aerobic capacity is the L-carnitine deficit on skeletal muscle. However, the value of the supplementation of L-carnitine to improve the physical fitness has been controversial. The objective of this work was to evaluate the effect of the administration of L-carnitine on VO2 max in hemodialyzed patients. PATIENTS AND METHODS: A group of 21 patients (20-50 years old) on a program of chronic hemodialysis was studied. During 12 weeks, 13 of them received L-carnitine, 7 men and 6 women, 38.8 (9.5) years old; BMI 24.2 (2.1) Kg/m2; 8 of them received placebo, 4 men and 4 women, 35.8 (11.4) years old; BMI 24.5 (5.8) Kg/m2. RESULTS: There was an increase in VO2 peak on L carnitine group from 16.3 (2.8) mL x Kg(-1) x min(-1) to 19.5 (3.3) mL x Kg(-1) x min(-1), and the same was seen in the placebo group (increase in VO2 peak from 14.8 (3.8) mL x Kg(-1) x min(-1) to 18.9 (4.8) mL x Kg(-1) x min(-1)). The L carnitine and placebo groups did not show statistical differences at the end of this study (all values above p > 0.05). CONCLUSION: In this group of patients, the intravenous supplementation of L-carnitine during 12 weeks did not have an impact on the improvement of the VO2 peak. PMID- 18405498 TI - [Factors related to the length of the corrected QT interval in patients treated with antipsychotics]. AB - BACKGROUND AND OBJECTIVE: The length of the heart-rate corrected QT interval (QTc) has been associated with an increased risk of cardiac dysrhythmia and sudden death. QTc length has been related to age, obesity, poor glycemic control, and use of drugs, such as antipsychotic medications. The objective of this cross sectional naturalistic study was to assess the factors associated with QTc length in patients treated with antipsychotics. PATIENTS AND METHOD: Bazett's formula for heart-rate correction was used to compute the corrected QT in 195 psychiatric inpatients treated with antipsychotics -117 males (60.0%), 78 females (40.0%); age (standard deviation): 28.4 (17.3) years (range: 12-84)-. RESULTS: Older age (p < 0.001) and female gender (p = 0.006) were the only factors significantly related to longer QTc interval. Diagnosis, type of antipsychotic, dosage of antipsychotic, tobacco use, and cardiovascular history were not related to QTc length. Only one male patient with QTc = 455 had a pathological QTc length (females > 470 ms, males > 450 ms). CONCLUSIONS: Factors related to QTc length in patients treated with antipsychotics are equivalent to those found in previous studies in patients not treated with antipsychotics. PMID- 18405499 TI - [Molecular characterization of heterozygous beta-thalassemia in Lanzarote, Spain]. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to determine the molecular defects of heterozygous beta thalassaemia and to ascertain their distribution in Lanzarote. PATIENTS AND METHOD: Molecular characterization was achieved by real time polymerase chain reaction (RT-PCR LightCycler, Roche), PCR-ARMS (PCR amplification reaction mutations system) and DNA sequencing on an automated DNA sequencer. RESULTS: Two hundred forty-three heterozygous beta thalassaemia carriers were included between July 1991 and February 2007. RT-PCR detected the molecular defect in 81% of the beta thalassaemia chromosomes analyzed [113 codon CD 39 (C --> T); 41 IVS-1-nt-110 (G --> A), 25 IVS 1-nt-1 (G --> A) and 19 IVS 1 nt-6 (T --> C)]. The remaining 12 molecular defects included the deletion 619 bp (7.8%) and the mutations -28 (A --> G), IVS1-nt-2 (T --> G), CD 41/42 (-TTCT), CD 8/9 (+G), CD 51 (-C), CD 22 (G --> T) and CD 24 (T --> A), CD 67 (-TG) and the novel mutation CD 20/21-TGGA. CONCLUSIONS: The distribution of the mutations is similar to that found in the Mediterranean area. The increasing migratory flow received in the Canary Islands may explain the emergence of new mutations not reported before in our area. PMID- 18405500 TI - [Statins and stroke]. PMID- 18405501 TI - [Hemoglobinopathy Newcastle: use of chromatography and first case reported in Spain]. AB - BACKGROUND AND OBJECTIVE: Unstable hemoglobins (Hb) show amino acid substitutions in critical places that produce a decrease of molecular solubility facilitating its denaturalization and precipitation. We describe the first case of Hb Newcastle in Spain. CASE REPORT: 5 year-old girl who came to visit due to fever over 4 days. Physical examination disclosed pale skin with subicteral mucosaes and splenomegaly. Lab analysis disclosed: Hb, 79 g/l; haematocrit, 0.27 l/l, mean corpuscular volume 93.4 fl, and reticulocyte count of 3%, along with anysocytosis and polychromasia. RESULTS: Hemoglobin heat stability test was positive. Hemoglobin electrophoresis showed a low band at HbA2. High performance liquid chromatography (HPLC) showed an Hb peak corresponding to the 12% of total Hb. Beta globin gene sequentiation showed the CD92 His --> Pro mutation Hb Newcastle in heterocygote condition in patient and her mother. CONCLUSIONS: Hb Newcastle has been described in 3 patients of English, Russian and Chinese origin. Clinical manifestation is chronic hemolytic anemia with severe crisis after oxidant drugs ingestion or infections. By using the electrophoretic method, a diffuse pattern of Hb bands between HbA and HbA2 is observed, difficulting the precise identification of the abnormal Hb. This inconvenience is overcomed by using HPLC that allows the clear identification of the abnormal Hb Newcastle. PMID- 18405502 TI - [Effectiveness of bisphosphonates in advanced prostate cancer]. PMID- 18405503 TI - [Effectiveness of drugs used in degenerative joint diseases (arthroses)]. PMID- 18405504 TI - [Metabolic syndrome, type 1 diabetes mellitus and insulin resistance]. AB - Although metabolic syndrome is generally associated with type 2 diabetes mellitus, the present review emphasizes the possible presence of metabolic syndrome in type 1 diabetes and the impact of insulin resistance on micro- and macrovascular complications. Results of major intervention studies in type 1 diabetes have led to a generalized intensive insulin treatment, being an excessive weight gain one of the consequences. Since abdominal obesity and hyperglycemia are 2 components of the metabolic syndrome, an increase in the percentage of metabolic syndrome in patients with type 1 diabetes is not surprising, though its prevalence does not reach that reported for type 2 diabetes. For insulin sensitivity quantification in patients with type 1 diabetes the estimated glucose disposal rate (eGDR): it consists of calculating a score based on clinical factors of the patient, which shows an inverse relationship with the development of micro- and macrovascular complications. Moreover, insulin resistance identification in type 1 diabetes may have therapeutic implications. PMID- 18405505 TI - [Multiculturality-oriented hospital. Experience in the Hospital del Mar, Barcelona, Spain]. PMID- 18405506 TI - [Screening for blood lead levels in children (corrected)]. PMID- 18405507 TI - [Reduction of hospital admissions after the development of a multidisciplinary program for cardiac failure in the internal medicine department]. PMID- 18405509 TI - [Pediatricians are key physicians for the eradication of cervix cancer]. PMID- 18405511 TI - [Tuberculous empiema necessitatis]. PMID- 18405510 TI - [Tophaceous gout causing stenosis of T11-T12]. PMID- 18405512 TI - [Fractional flow reserve in diabetics: does it have an expiry date?]. PMID- 18405513 TI - [Is acute myocardial infarction an appropriate setting for drug-eluting stents?]. PMID- 18405514 TI - [Cardiac surgery in the cancer patient]. PMID- 18405515 TI - [Long-term prognosis in diabetic patients in whom revascularization is deferred following fractional flow reserve assessment]. AB - INTRODUCTION AND OBJECTIVES: The fractional flow reserve (FFR) has been shown to be a valid and useful measure in the functional assessment of coronary stenoses of intermediate severity. Our aim was to determine the usefulness of FFR assessment in diabetic patients, in whom determination of the FFR can be influenced by microvascular dysfunction. METHODS: Between 1997-2004, FFR assessment was used to evaluate 222 consecutive coronary lesions judged by an interventional cardiologist to be of intermediate severity (ie, 40%-70%). Intravenous adenosine (140 microg/kg per min) was used to achieve maximum hyperemia. The occurrence of cardiac events (ie, death, non-fatal acute myocardial infarction, and target lesion revascularization) was compared in diabetics and nondiabetics in whom FFR assessment gave a negative result and intervention was deferred. The mean follow-up period was 30+/-21 months. RESULTS: Revascularization was deferred for 144 lesions (in 136 patients) in which the FFR was >/=0.75. Of these, 42 lesions (29.2%) were in diabetics (40 patients). The proportion of patients who were female or who had hypertension, dyslipidemia or multivessel disease was greater in the diabetic group. There was no difference in indications for coronary angiography. In both groups, the most frequently investigated vessel was the left anterior descending coronary artery. The mean FFR was 0.87+/-0.06, and there was no difference between the groups. On long-term follow-up, there was no difference in the rate of death or acute myocardial infarction. Overall, 8.8% of nondiabetics and 14.3% of diabetics with a negative FFR test result required target lesion revascularization (P=.32). CONCLUSIONS: Our results indicate that deferring percutaneous coronary intervention in diabetics with a moderately severe coronary artery stenosis and an FFR >/=0.75 is safe. PMID- 18405516 TI - [Long-term clinical and angiographic follow-up of drug-eluting stents in patients with ST-elevation acute myocardial infarction]. AB - INTRODUCTION AND OBJECTIVES: Drug-eluting stents (DES) have proven to be effective in reducing the rate of restenosis and have, therefore, been incorporated into the treatment of patients with ST-elevation acute myocardial infarction (STEMI). The aim of this study was to investigate long-term clinical and angiographic outcomes following the use of DESs in patients with STEMI. METHODS: A prospective study involving clinical and angiographic follow-up was performed in 81 patients with STEMI who underwent percutaneous coronary intervention including DES implantation. This group was compared with 82 patients with similar characteristics who were treated with bare-metal stents (BMS) in an earlier period. RESULTS: At one year, there was no significant difference between the groups in the mortality (2.5% in the DES group vs 7.3% in the BMS group; P=.15) or reinfarction rate (4.8% in the DES group vs. 4.8% in the BMS group; P=.98). The target lesion revascularization rate was significantly lower in the DES group (8.6% vs 23.2% in the BMS group; P=.001), as was the restenosis rate (13.8% vs. 30.9% in the BMS group; P=.02). Acute or subacute stent thrombosis was diagnosed in five patients (3 with a DES and 2 with a BMS; P=.64), and one late stent thrombosis was detected after a year, in a sirolimus-eluting stent. CONCLUSIONS: Implantation of a DES in patients with STEMI did not result in a reduction in either the mortality or reinfarction rate at 1 year compared with BMS implantation. However, there were reductions in the rates of restenosis and target lesion revascularization. The incidence of thrombosis was similar with the two types of stent. PMID- 18405517 TI - [Cardiac surgery with extracorporeal circulation in cancer patients: influence on surgical morbidity and mortality and on survival]. AB - INTRODUCTION AND OBJECTIVES: Morbimortality related to cardiac surgery may be superior in patients with malignant neoplastic disease. Inflammatory phenomena and immunologic changes secondary to extracorporeal circulation use can also increase tumor recurrence. We evaluate characteristics and results of cardiac surgery in our neoplastic patients. METHODS: Out of 2146 consecutive patients who underwent cardiac surgery with extracorporeal circulation, 89 (4.2%) had been previously affected by cancer. Cancer was active (recent diagnosis or under treatment) in 33 patients (group A) and 56 (group B) were in remission. Both groups were matched with 165 patients with no tumor, according to age, gender, type of surgery, and comorbidity (group C). We retrospectively evaluated incremental risk factors for surgical morbimortality, survival and tumor recurrence. RESULTS: Median interval between cancer diagnosis and surgery was 60 months and mortality and morbidity were 4.5% and 36%, respectively, vs 5,4% and 32,7% in group C. During follow-up, 12 patients died (8 due to cancer), 16 suffered cancer recurrence and 2 new tumors were diagnosed. Statistical analysis did not permit us to identify any incremental risk factor for mortality. Postoperative morbidity was increased in case of preoperative renal failure. During follow-up, survival was significantly decreased in group A, in case of preoperative left ventricular dysfunction or pulmonary obstructive disease, and when interval between cancer diagnosis and cardiac surgery was under 2 years. CONCLUSIONS: We have not observed an increase in cardiac surgery morbimortality in cancer patients. Anyway, survival is decreased in case of active or recently diagnosed cancer. PMID- 18405518 TI - [Coronary artery bypass surgery using the mini-extracorporeal circulation system: a Spanish unit's experience]. AB - INTRODUCTION AND OBJECTIVES: The increasing use of percutaneous interventions has resulted in a significant reduction in coronary artery bypass grafting. Today, patients referred for surgery are older, have more comorbidities, and have undergone previous percutaneous intervention, and their ventricular function is poorer. As a result, surgery has attempted to improve its results by adopting a number of different strategies. The aim of this study was to investigate and describe one cardiac surgery unit's initial experience with coronary artery bypass grafting using mini-extracorporeal circulation (MECC), which had become its technique of choice. METHODS: A retrospective analysis of 408 patients who underwent isolated coronary artery bypass grafting using MECC between January 2004 and April 2007 was carried out. Of the 408, 329 (80.6%) were men, their mean age was 63.5 years (28-83 years), 63% had hypertension, 49.3% had diabetes, 69% had hyperlipidemia, and 52% were smokers. RESULTS: The surgical mortality rate predicted by the logistic EuroSCORE was 3.7% (range, 1-38). Overall, 34% of patients had left main coronary artery disease and 87% had three-vessel disease. In 74%, complete revascularization was carried out using a mean of 2.97 (range, 1 7) grafts per patient. A mammary artery graft was used in all cases. The in hospital mortality rate was 0.74%. There were few postoperative complications: 0.98% of patients required further surgery because of bleeding, 3.4% had a significantly elevated troponin-I level, 6.4% developed kidney failure, and 0.5% suffered a stroke. CONCLUSIONS: Coronary artery bypass grafting using MECC enabled complete revascularization to be performed in most patients, and morbidity and mortality rates were low. PMID- 18405519 TI - [Cost-effectiveness of managing familial hypercholesterolemia using atorvastatin based preventive therapy]. AB - INTRODUCTION AND OBJECTIVES: A cost-effectiveness model was developed to evaluate the efficiency of different preventive strategies in familial hypercholesterolemia (FH) in comparison with routine clinical practice (CP): atorvastatin monotherapy, 40 mg (A40) or 80 mg (A80, and atorvastatin combined with ezetimibe, 10 mg (A40+E10 or A80+E10). METHODS: A longitudinal population model with a time horizon for life-expectancy was developed within the context of the Spanish public healthcare system. Life tables for the Spanish population (2002) were modified using the standardized mortality rate for individuals with FH. Effectiveness was expressed in life-years gained (LYG), after taking into account reductions for risk (ie, Framingham risk score) and cardiovascular mortality. The costs (in 2005 terms) of the intervention (CI) and care (CC) were discounted at 6%, while effects were discounted at 3%. RESULTS: Routine CP, based on the Spanish FH registry: 1.97 LYG per patient vs. no treatment; CI euro5321, CC euro23,389. A40: 2.59 LYG; reduction in CC compared with CP 4.5%; total costs (TC) euro30 569. A80: 2.75 LYG; reduction in CC 6.4%; TC euro30 133. A40+E10: 3.38 LYG; reduction in CC 14.3%; TC euro36 104. A80+E10: 3.62 LYG; reduction in CC 17.6%; TC euro35 317. From most to least efficient strategy, the incremental cost-effectiveness per LYG compared with CP was: a) A80: euro1821; b) A40: euro3012; c) A80+E10: euro4021, and d) A40+E10: euro5250. CONCLUSIONS: Preventive treatment of FH with atorvastatin was cost-effective. The greatest cost effectiveness was obtained with atorvastatin monotherapy, 80 mg. The addition of ezetimibe could produce further benefits at an acceptable incremental cost. PMID- 18405520 TI - [Changes in the spectral characteristics of ventricular fibrillation in lesions produced by radiofrequency ablation. An experimental study]. AB - INTRODUCTION AND OBJECTIVES: Although electro- physiological databases contain information about changes in the time domain in lesions produced by radiofrequency ablation, very few data on changes in the frequency domain are available. The aim of this study was to investigate changes in the spectral characteristics of ventricular fibrillation in zones with radiofrequency lesions. METHODS: Recordings of ventricular fibrillation were obtained in 11 isolated perfused rabbit heart preparations using a multiple epicardial electrode located on the left ventricular free wall. Spectral parameters derived by Fourier analysis before and after the creation of transmural radiofrequency lesions were compared. RESULTS: In the ablated zones, significant reductions were observed in the spectral density of the dominant (0.168+/-0.113 mV(2)/Hz vs 0.025+/-0.018 mV(2)/Hz; P< .001) and mean frequencies (0.053+/-0.057 mV(2)/Hz vs 0.012+/-0.016 mV(2)/Hz; P< .001), the normalized energy around the dominant frequency (0.860+/ 0.570 vs 0.128+/-0.091; P< .001), and the standard deviation of the power spectrum (0.031+/-0.020 mV(2)/Hz vs 0.004+/-0.001 mV(2)/Hz; P< .001). There was no significant change in the dominant (16.2+/-5.6 vs 14.8+/-1.8 Hz) or mean frequency (17.7+/-3.4 vs 16.6+/-1.3 Hz). The spectral parameters that could be used in a multivariate model to identify the lesion were the standard deviation of the power spectrum and the spectral density of the mean frequency. CONCLUSIONS: During ventricular fibrillation, the spectral parameters associated with spectral power and spectral energy were significantly altered in zones with radiofrequency lesions and could be used to identify those zones. There was no significant change in either the dominant or mean frequency in these zones. PMID- 18405521 TI - [Risk functions and the primary prevention of cardiovascular disease]. AB - Cardiovascular risk functions are regarded as the best tools for establishing priorities in primary prevention. Since the original Framingham risk chart fell into disuse because it greatly overestimated the real risk, the adjusted REGICOR and SCORE functions have become widely available in Spain, although the REGICOR function is the only one that has been validated for use in the Spanish population. Risk estimates have been shown to be useful for decision-making, particularly on the treatment of hypercholesterolemia. However, the fact that the majority of cardiovascular events occurs in individuals classified as being at a medium risk is evidence for the poor discriminative ability of classical risk factors. Despite the use of new parameters proposed for estimating cardiovascular risk, such as the C-reactive protein level, the detection of coronary calcification, the carotid intima-media thickness and the ankle-brachial index, there has been no improvement in the predictive capacity of classical risk factors. The most promising alternative seems to be the identification of "vulnerable patients" using markers of vulnerable plaque (ie, unstable or high risk plaque), vulnerable blood (ie, with a tendency for thrombosis), and vulnerable myocardium (ie, electrically unstable or with a tendency for arrhythmia). In this article, we discuss whether the combined use of cardiovascular risk functions, novel risk markers and noninvasive tests can be effective in increasing the accuracy of patient selection for the primary prevention of cardiovascular disease. PMID- 18405522 TI - [Infective endocarditis of the interatrial septum due to a mitral regurgitant jet]. PMID- 18405523 TI - [Prevalence of primary aldosteronism in hypertensive patients and its effect on the heart]. AB - Primary hyperaldosteronism (PHA) is thought to have a harmful effect on the cardiovascular system and, in recent years, the number of cases of hypertension due to PHA has been increasing. The aims of this study were to determine the prevalence of PHA and to assess cardiac damage associated with the condition in 183 consecutive hypertensive patients. A full secondary hypertension work-up was performed, and included ECG and echocardiography. In total, 11 (6%) patients were diagnosed with PHA. Compared with other hypertensives, those with PHA had higher systolic blood pressure, more frequently had evidence of left ventricular hypertrophy on ECG (45.5% vs 11.6%; P< .01), and had a larger left ventricular mass on echocardiography (145.5 g/m(2) vs 97.52 g/m(2); P< .0001). In conclusion, PHA is a significant contributor to the increasing prevalence of hypertension and its effect on the heart is greater than that of other causes of hypertension. PMID- 18405524 TI - [Benefits of cardiac resynchronization therapy in patients with atrial fibrillation who have not undergone atrioventricular node ablation]. AB - The aim of this study was to compare the effects of cardiac resynchronization therapy on left ventricular function and reverse remodeling in patients in sinus rhythm with the effects in patients with atrial fibrillation who have not undergone atrioventricular node ablation. Echocardiographic and clinical parameters were evaluated at baseline and after 6 months of cardiac resynchronization therapy in 55 patients: 15 had atrial fibrillation and 40 were in sinus rhythm. Device programming was similar in the 2 groups, as were the reductions in QRS interval and echocardiographic measures of asynchrony observed after implantation. However, although significant improvements in end-systolic volume and ejection fraction were seen in both groups, reverse remodeling was greater in patients in sinus rhythm (reduction in end-systolic volume 30.9%+/ 24.6% vs 12.5%+/-18.6%; P=.024), as was the relative increase in ejection fraction (15.4%+/-12.6% vs 5.0%+/-7.2%; P=.010). Cardiac resynchronization therapy in patients with atrial fibrillation who had not undergone atrioventricular node ablation resulted in significant improvements in ejection fraction and reverse remodeling, but these were less than those observed in patients in sinus rhythm. PMID- 18405525 TI - [Usefulness of the ECG in the sports screening of footballers affiliated to a regional sports federation]. AB - We report the results of a study carried out in 825 young football club members, all of whom underwent screening for hypertrophic cardiomyopathy using a 12-lead ECG. Echocardiographic assessment was performed in only those with positive ECG results, as defined by the European Society of Cardiology. Echocardiography proved necessary in 61 (7%) individuals with positive ECG findings, of whom 7 had echocardiographic findings indicative of left ventricular hypertrophy: five were in the "gray zone", 1 was judged to have athlete's heart, and one had been diagnosed with hypertrophic cardiomyopathy. In all these cases, ECG showed repolarization abnormalities, and 4 satisfied criteria for left ventricular hypertrophy. PMID- 18405526 TI - [Posterior myocardial infarction is real]. PMID- 18405528 TI - [Pheochromocytoma-related cardiomyopathy or stress cardiomyopathy secondary to pheochromocytoma: is new terminology needed?]. PMID- 18405529 TI - [Iatrogenic coronary-cameral fistula after percutaneous intervention on the right coronary artery]. PMID- 18405530 TI - [El Camino de Santiago in the service of cardiovascular rehabilitation]. PMID- 18405531 TI - [A hybrid approach to hypoplastic left heart syndrome]. PMID- 18405532 TI - [Recommendations of the Analgesia and Sedation Group of SEMICYUK for sedation/analgesia and neuromuscular blocking in the critical patient. Presentation]. PMID- 18405533 TI - [Intubation of the critical patient]. AB - The airway management is one of the principal skills that a physician needs to ensure optimal ventilation and oxygenation. In this guideline, Sedation and Analgesia Working Group of SEMICYUC describes rapid sequence intubation (RSI) and induction drugs and neuromuscular blocking agents. RSI is the best procedure to ensure optimal airway management in the majority of critically ill patients. Our choice of one induction drug or another can influence in the success of the airway management. As neuromuscular blocking agents can facilitate intubation, they influence the choice of the drug for intubation and of premedication. To optimize the use of drugs, the knowledge of pharmacodynamics, pharmacokinetics and side effects is imperative. A proper position of the patient is essential to establish an adequate airway management. Direct visualization of glottis and endotracheal tube pass through vocal cords is the best way to confirm the correct position of it. There are different devices to confirm correct position of the endotracheal tube. PMID- 18405534 TI - [Short duration sedation]. AB - One of the main factors to keep in mind for drug selection for the sedation of a critical patient is its foreseen duration. We have denominated by consent, short sedation that whose duration is less than 72 h. We advise the existence and application of a protocol of short sedation in all intensive care units. This protocol must guarantee the patient's analgesia at all times and its application must not increase the times of waking up and weaning of the mechanical ventilation. In this chapter the pharmacology and the comparative studies of the drugs more used for this aim are revised and the clinical recommendations are settle down. Some recommendations for specific situations are also settle down and a role is assigned to less habitual drugs such as ketamine. PMID- 18405535 TI - [Prolonged sedation in Intensive Care Units]. AB - Sedation and analgesia constitute one of the cornerstones in the management of the critically ill patients. Most patients admitted to an Intensive Care Unit require prolonged sedation and analgesia. It has been demonstrated that adequate sedo- analgesia lessens stress-related events in the critically ill patients, facilitating their management and improving their outcomes. However, the use of sedatives and analgesics, especially when administered in continuous intravenous infusion, may have its complications derived from its infra or over utilization with proved impact on the outcome of critically ill patients. A proper monitoring and the implementation of sedation and analgesia protocols warrant the adequate management of existing sedatives aiding to avoid tolerance and dependency events. Strategies such as "sequential sedation", "dynamic sedation" or "daily sedation interruption" have been proposed as efficacious tools for the avoidance of complications related to prolonged sedation. In the present chapter, concepts related to prolonged sedation (meaning sedation for more than 72 hours) are reviewed; available agents are evaluated and strategies aimed to assure quality in its application are described. PMID- 18405536 TI - [Control strategies for difficult sedation]. AB - There is a wide intra- and inter-individual variability in sedative dose requirements in mechanically ICU patients. Patient's heterogeneity, the frequent and variable organic dysfunctions, the drug interactions and the possibility of metabolite accumulation could explain this variability. However, this fact must not justify the use of excessive doses to achieve the goals of sedation. Frequently, in the absence of a specific motive, e.g. uncontrolled pain, physicians administer progressive sedative dose increases. Probably, the absence of maximum dose recommendations has originated the case's description of severe complications, sometimes mortal, or sedative toxicity, like propofol infusion syndrome. This SEMICYUC Analgesia and Sedation Work Group recommends not administering more than 4.5 mg/kg/h of propofol or 0.25 mg/kg/h of midazolam. The need to use more than these doses should force a change in the sedative or the combined administration of both. Depending on the clinical situation or the clinical patient's evolution, the use of clonidine, haloperidol or remifentanil could be better options. PMID- 18405537 TI - [Monitoring of pain. Recommendations of the Analgesia and Sedation Work Group of SEMICYUC]. AB - In critically ill patients, pain is frequently underestimated and so insufficiently managed. Psychological, haemodynamic and neuroendocrine responses, secondary to untreated pain, could produce morbidity and even increases in patient mortality. All members of the intensive care team must have abilities to assess and to manage pain. The evaluation of pain in the critically ill patient is very difficult but extremely important. Self-reported pain is the starting point for treatment. The pain scores recommended are, VAS (visual analogue scale) and NRS (numeric rating scale) in communicative patients and Campbell scale in uncommunicative patients. Adequate and regular patient assessment leads to improved pain control. Scores higher than 3 points should not be permitted. A pain-free Intensive Care Unit should be a quality standard healthcare aim. PMID- 18405538 TI - [Monitoring of sedation]. AB - Once analgesia is assured, sedation has special relevance in the critically ill ventilated patient's global treatment. Sedatives should be adjusted to individual needs, by administering minimal effective doses to achieve the AIM. This aim must be clearly identified, defined at the beginning of the treatment and revised on a regular basis, ideally at least once per shift. Sedation strategies should foresee the different needs throughout the day within dynamic sedation concept framework. Required sedation depth depends on the patient's psychological characteristics, foreseen evolution and patient tolerance to the support techniques used in treatment. Sedation monitoring permits identification and correction of under- or over-sedation, either of which could negatively influence critically ill patient evolution. The over-sedation concept must be applied to all situations where patients receive more sedation than required. This Spanish Society of Critical Care Medicine's Analgesia and Sedation Work Group recommends the Richmond Agitation Sedation Scale, due to its interrelationship with the Confusion Assessment Method Scale (CAM-ICU), for sedation monitoring in patients under light sedation while it recommends bispectral index sedation monitoring in patients under deep sedation. In the latter case, maintaining values under 40 on the bispectral index doesn't produce any benefits except in patients who require a maximum decrease in neuronal metabolism. To avoid recall phenomena, bispectral monitoring is highly advisable in patients treated with neuromuscular blockers. PMID- 18405539 TI - [Monitoring of neuromuscular blocking]. AB - Neuromuscular blockade monitoring aims should be based on effectiveness criteria, that is, to administer the lowest effective dose, and on security criteria, avoiding overdosage and detecting possible residual blockade before patient extubation. A neuromuscular blockade monitoring and usage protocol should be available with predefined objectives for each patient to achieve the minimum effective doses. Maintenance of a light blockade level probably influences the decrease in complications associated with these drugs' use. Train-of-four monitoring, along with muscular movement detection allow adjustment to meet these aims. Ulnar nerve stimulation with response detection in the adductor pollicis muscle is the recommended neurostimulation in critically ill patients, while facial neurostimulation is recommended in situations where edema or skin temperature prevent correct ulnar nerve stimulation. Ensuring adequate sedation and analgesia in a paralysed patient is essential. An inadequately sedated but paralysed patient may subsequently suffer serious psychological and emotional stress. Bispectral index monitoring with sedative doses adjusted to 40-60 values assures, in most situations, correct sedation. PMID- 18405540 TI - [Use of neuromuscular blockers in the critical patient]. AB - The use of neuromuscular blockers (NMB) is a frequent practice in Intensive Care Units. However most of the experience with the use of these agents come from the operating room used to deal with patients with different characteristics from those admitted in the ICU. Recent advances on neuromonitoring and the commercialization of newer agents make necessary the update in the management of NMB in the ICU. The NMB agent should be chosen attending to its pharmacokinetics and the physiopathology of the critically ill patient. Those NMB with organ independent metabolism as well as those with rapid onset of action are the preferred ones for the use in the critically ill patient substituting older depolarizing agents and those whose metabolism is dependent on the liver and/or kidney, organs frequently impaired in the critically ill patients. Neuromuscular blocking in the critically ill patient should be done according to protocols and monitor its effects in order to avoid complications related to its prolonged use. PMID- 18405541 TI - [Delusion in the critical patient]. AB - Delirium, the acute confusional syndrome, is a common although infradiagnosed problem in the critically ill patient, especially the hypoactive subtype. Risk factors for delirium are previous cognitive disturbances, some comorbidities, ambiental factors and the acute organic alterations of critical illness. Delirium is associated to an increase in short and long term mortality, prolongation of mechanical ventilation, increased Intensive Care Unit (ICU) and hospital length of stay, and cognitive impairment after hospital discharge. In the last years several tools have been developed to detect delirium in critically ill patients. The Intensive Care Delirium Screening Checklist (ICDSC) and the Confusion Assessment Method for ICU patients (CAM-ICU) have been validated and are useful even in patients receiving mechanical ventilation. Some interventions on specific risk factors can decrease the incidence of delirium in hospitalized patients. Treatment of delirium is based on the identification and correction of contributing factors, the introduction of support measures, and pharmacological therapy for symptomatic control. Halloperidol is the first line therapy of delirium in the critically ill patient, while experience with atypical neuroleptics and other drugs is limited, precluding to do recommendations about its use. Neuroleptic drugs can produce severe side effects and need careful dosage and monitoring. When agitation is important, can be necessary the simultaneous use of benzodiazepines or propofol, and some times, the temporal and protocolized application of physical restraints. PMID- 18405542 TI - [Depression and use of antidepressants in the critical patient]. AB - A large percentage of critically ill patients suffer from depression while admitted in an Intensive Care Unit (ICU). This pathology, often underdiagnosed by intensive care professionals has a proved negative impact on median-large outcome, which makes early detection and management a key issue. However diagnosing depression in ICU is a complicated task since there are no validated tools for its detection. The cornerstone intervention for the treatment of depression are antidepressant medication. All antidepressants have similar efficacy profiles. The prescription of a particular agent should be done based in its collateral effects. Unfortunately the efficacy and safety of antidepressant agents has not been evaluated in the critically ill patient. The implementation of simple measures like guaranteeing comfort during its admission to the ICU and the early reintroduction of any psychotropic medication that the patient could be taking before ICU could improve the emotional adaptation to their new situation. PMID- 18405543 TI - [Loco-regional analgesia]. AB - The epidural analgesia is one of the most effective techniques for pain relief when it is indicated, but it can present potentially serious complications that must precociously be diagnosed and be treated. In the Critical Care setting, epidural analgesia is used for pain control after surgery or major trauma. The technique is simple, a catheter is placed into a virtual cavity, so the administered drugs are absorbed through the epidural space into nerve roots. The administration of local anesthetics, opioids or the combination of both by epidural route (administered in continuous infusion or bolus), provides better analgesia. Also the clonidine can be used. In order to diagnose and to treat suitably the possible complications (pain, urinary retention, nauseas and vomits, itching, motor block, infection, respiratory depression, hypotension) a series of safety measures must be adopted (respiratory and heart rate, blood pressure, sedation score, sensory and motor level assessment, rate of diuresis, temperature and signs of infection). PMID- 18405544 TI - [Sedation in special procedures and situations]. AB - Numerous diagnostic techniques require sedation and analgesia in order to be performed in a safe and comfortable way for the patient. Several of the most notable points of interest for the critical care specialist are the electrical cardioversion, the placing of implantable cardiac stimulation devices, the endoscopic techniques and the performing of bedside surgical procedures. In this current revision, the SEMICYUC Task Force for Sedation and Analgesia describes recommendations and best practices for administering sedation and analgesia in these situations. PMID- 18405545 TI - [Analgesia, sedation and relaxation in the child with mechanical ventilation]. AB - The basic concepts of sedation and analgesia and the tools to asses the level of sedation and analgesia are review. The different methods of sedation and the non pharmacological interventions are described. Sedatives, analgesics and muscle relaxants, their pharmacodynamics and pharmacokinetics in children, their indications in specific situations (intubation, pain control, sedation and neuromuscular blocking) are reviewed. The etiology of patient-ventilator asynchrony in ventilated children and how to treat it are analyzed, giving guides of how to adapt sedation to the level of mechanical ventilation therapy. Finally, general recommendations are given for the analgesia and sedation in mechanically ventilated children. PMID- 18405546 TI - [The seven deadly sins of impact evaluations. 2008 SESPAS Report]. PMID- 18405547 TI - [The role of epidemiology in policy definition. Toward the public health required. 2008 SESPAS Report]. AB - Epidemiology already plays a substantial role in the definition of public and corporate policies in Spain as well as in other developed countries. The World Health Organization reinforced the position of epidemiology by enshrining the notion of social health determinants and, as a result, the connection between health and social constructs has become more visible. However, the use of epidemiology is usually restricted to the development and evaluation of policies originating in health bureaucracies with health improvement as the only public objective; these are called direct health policies. This view is somewhat reductionistic insofar as health gains and losses occur mainly due to policies outside the health sector. To expand the role of epidemiology as a tool to shape policies, we need a view of exposures and effects beyond healthcare and the biological and behavioral risk factors proposed by medical paternalism. Also required are an attitude that is more participative than prescriptive and the development of more health impact assessments. In addition, the health effects of the decisions taken by politically-empowered institutions should be included in the epidemiologist's tool kit. In other words, a political epidemiology should be constructed. For this endeavor, which may be crucial to attain the public health required, public health practitioners should be more visible and credible, have greater independence from political power, provide information directly to the public and other stake holders (greater transparency), and further develop the professionalization of public health (greater competence) by working increasingly in networks of professionals rather than in bureaucratic pyramids. PMID- 18405548 TI - [Integration of information for health interventions: from data to information and from information to action. 2008 SESPAS Report]. AB - Spanish public health and health services information systems (HIS) have improved, but are still fragmented by areas of interest and have evolved independently from one another. Their format, structure, integrity and data quality vary widely, as do the programs, platforms and databases that support them. The latest innovations focus on isolated HIS and are hampered by obsolete models, tools, functionalities, and the inertial demand of information. Transfer of responsibilities without minimal agreements on HIS has eroded their national cohesion and, along with the absence of exchanges on experiences of computerization on a national basis, has weakened us, given the supply of immature computer applications. The evolution of HIS must be governed by integration. We have to redefine their strategic and operational objectives, review existing data and information, and determine the single identification of specific persons and patients. Variables, indicators, services and control panels should be reviewed and systematized through a single shared nomenclature. Personal health records and administrative and clinical registries should become the primary sources of health information data. Data collection, mechanization, registration and exploitation, and their quality control and maintenance, should be redefined regardless of setting. A national agreement is urgently required on the minimal functionalities of HIS, while respecting their technical nature and management by autonomous governments. PMID- 18405549 TI - [How can we improve effectiveness (quality) by reducing the gap between research and action? 2008 SESPAS Report]. AB - In our environment, research on health technologies and health care effectiveness is a prominent issue in current knowledge policies. Nevertheless, there is wide consensus on the need for additional efforts to develop the translational component of this kind of research under the hypothesis that taking research into practice is a necessary condition for improving healthcare. The characteristics that facilitate or hinder knowledge transfer in healthcare organizations are analyzed throughout the present text. At the same time, we discuss some successful experiences that illustrate how the chasm can be reduced and healthcare effectiveness (i.e. quality) improved. Among the mechanisms that might either reduce or maintain the chasm are the following: 1) the type of organization (healthcare provider model, teaching status, mix of professionals, the organization's capacity to learn and innovate, etc.); 2) the organization's government (leadership in evidence implementation, capacity to manage professional roles, ability to use economic incentives, availability of an information-rich context, the use of knowledge management, etc.), and 3) the effectiveness of the strategies themselves in translating evidence into practice. Finally, we describe some successful experiences focusing on implementation, on researching the evidence that is needed, and on reinventing the organization to convert information into knowledge. PMID- 18405550 TI - [Marketing as a tool to increase the effectiveness of public health plans. 2008 SESPAS Report]. AB - Recent years have seen a steady increase in social marketing applied to health with the aim of increasing public awareness and changing people's behavior. Programs or actions based on the principles of social marketing have been shown to be effective in improving public health. However, that is not the general rule in Spain, where health policies have been based on health plans directed more to economic efficiency than to citizens' needs. For a health marketing program to be effective, the following factors are necessary: 1) the program has a long-term temporal horizon for action; 2) the objectives are established in terms of behavioral changes; 3) market research is used as a source of information; 4) different actions are established according to the segments identified as targets; 5) the program is operationalized in the four variables of the marketing mix, namely, product, price, distribution and communication, and is not only based on advertising campaigns; 6) the core of the program is exchange, understood as the factors that motivate people to change in return for the promise of something beneficial to them; 7) the factors or forces that compete with the desired behavioral changes are neutralized, and 8) businesses' social responsibility is used as a mechanism to reinforce health improvement programs. The design of health marketing programs should include definition of strategic and operational actions aimed not only at potential adopters of the desired behavior but also at all agents who may help or hinder behavioral change (health professionals, the pharmaceutical industry, politicians, the advertising industry, and products and services with health-challenging objectives). PMID- 18405551 TI - [The new physician. A pressing reform in medical education. 2008 SESPAS Report]. AB - The reform of medical education is an active process worldwide. This document focuses on the situation in Spain in analogy with other reference countries. We analyze the structure and functioning of the sequential stages of medical education and training: undergraduate education, specialty training and continuing professional development. We also analyze the Bologna process, the numerus clausus, traditional and new academic programs, residency programs for medical specialization, continuing medical education, professional advancement and specific licensing areas. PMID- 18405552 TI - [Health expenditure in the context of social expenditure. A generational analysis of tendencies in Spain in the context of demographic aging. 2008 SESPAS Report]. AB - The tendency for public welfare spending to be increasingly aimed at the elderly has been identified in several developed countries. While population aging is a common trend, it is not obvious why the shift in spending exceeds the trend in aging, or why per capita spending on the elderly is increasing. In the first part of the present article, we show that this trend is occurring in Spain, identify the losers from this development, discuss the policies that underlie it, and propose adjustments based on Musgrave's fixed proportions rule for fair intergenerational distribution. These policies aim to manage population aging, labor market participation of youth and women, as well as public policies that combine 'work-fare' strategies with the more traditional 'welfare' strategies. In the second part of this paper, we explore the contribution of public health expenditure to overall public social expenditure, and analyze the effect of increasing health on distributional fairness. This analysis is guided by our perception that social policy, including health policy, should be more horizontal, i.e. it should take into account the sum total of a mixed basket of resources aimed at any recipient group, to avoid the skewed allocation of resources that arises from compounding various independent distributions of resources. Coordination of the various channels of social expenditure should ensure precisely targeted recipients and cross-departmental sources. The normative fairness criterion to be used should be precisely specified. In Spain (1980-2000) the oldest segment of the population has gained the most, appropriating an increased share of resources. This increase goes beyond the amount that could be explained by population aging throughout the period. The 'losers' are individuals with different fragility conditions. On average the youngest working group has suffered the highest relative loss. PMID- 18405553 TI - [Transport, environment and health. 2008 SESPAS Report]. AB - Motor road transport has increased exponentially in the last few years. To a large extent, mobility is an essential element in the organization of society, but, until recently, the implications of chosen forms of transport for health and the environment have not been considered. In this chapter we review the negative impact of current forms of transport on health in terms of traffic injuries, climate change, atmospheric contamination, noise, and interference with daily activities and exercise, such as impediments to walking or cycling. Some possible interventions related to the instruments available in public health and other fields are proposed. Issues deserving further research are highlighted. Some examples in Spain and other countries are described. Recommendations are made on the need to reduce the use of private cars and to develop segmented routes and areas of quiet traffic connected in the cities and among nearby towns to promote walking are cycling. One major goal in current public policies should be to develop and maintain a public transport system that is safe, cheap and faster and less polluting than private transport. These interventions would help to achieve a change in current modes of transport and would lead to a healthier population and a more sustainable environment. PMID- 18405554 TI - [Health threats and health system crises. An approach to early warning and response. 2008 SESPAS Report]. AB - The world is changing more and faster than ever before. New diseases are coming to light each year, controlled diseases are reemerging as potential threats, and natural or man-made disasters are increasingly affecting human health. The "International Health Regulations (2005)" reflect the changes in the response of public health to this new situation. Surveillance of specific diseases and predefined control measures have been replaced by surveillance of public health events of international concern and control measures adapted to each situation. The public health events of international interest are characterized by their seriousness, predictability, the risk of international spread and potential for travel or trade restrictions. The development of the European Early Warning and Response System in 1998 and the creation of the European Center for Disease Prevention and Control in 2005 demonstrate political commitment in Europe, with early detection of and response to public health threats. However, timely risk evaluation and response at a national level requires improved data digitalization and accessibility, automatic notification processes, data analysis and dissemination of information, the combination of information from multiple sources and adaptation of public health services. The autonomous regions in Spain are initiating this adaptation process, but interoperability between systems and the development of guidelines for a coordinated response should be steered by the National Interregional Health Council and coordinated by the Ministry of Health. Efficient early warning systems of health threats that allow for a timely response and reduce uncertainty about information would help to minimize the risk of public health crises. The profile of public health threats is nonspecific. Early detection of threats requires access to information from multiple sources and efficient risk assessment. Key factors for improving the response to public health threats are the development of surveillance methods and operational research in public health. PMID- 18405555 TI - [The health of cities and their citizens (urban development and municipal public health). 2008 SESPAS Report]. AB - The policy of continued improvement in citizens' health, further to the definitions provided in the latest World Health Organization reports, entails a huge undertaking on the part of local administrations in issues that not only affect health management but also involve a change in other health-related policies, such as urban development and planning. Until now, urban development has been reluctant to incorporate the criteria defined by health strategies into its field of knowledge. However, there is a lack of research in public health, which could specify the clear effects of the environmental on citizens' health and enable criteria applicable to spatial planning to be defined. The present article reflects on certain experiences acquired in our environment of processes that bring together these two views of health, ranging from urban and environmental policies in indicator-based reports to a series of good practice. The integrated municipal action of San Fernando de Henares in the Healthy Cities project has also been significant. PMID- 18405556 TI - [Regulatory policies on alcohol in Spain. Experience-based public health? 2008 SESPAS Report]. AB - The present article reviews the proposal for alcohol regulation made in Spain in 2006 by the Ministry of Health and Consumer Affairs and dropped a few months later as adverse positions took over the debate in the political arena and the media. The background to these regulations, as well as their components, the process, and the actors involved are analyzed, and the factors leading to this outcome are discussed. A comparison is made with the tobacco regulation initiative in 2005, which resulted in a regulatory law with a highly favorable impact on public health. The actors interested in promoting alcohol consumption and opposed to any regulation have a privileged institutional presence, generating powerful resistance. Although these regulatory proposals would have marginal impacts on their trade, wine growers and wineries have been the most visible forces against regulation and have had the greatest political and media impact. Equally, manipulated messages on the health benefits of alcohol use have fed arguments against regulation. The lack of political and media consensus in this case stands in contrast with the tobacco regulation process, in which a certain political consensus had previously been reached and the smoking prevention movement had permeated the media with preventive messages, framing the issue in terms favorable for regulation. To gain ground, the sectors interested in expanding public policies for the prevention of the harm caused by alcohol need greater cohesion and organization. Professional public health organizations can substantially contribute to this field. PMID- 18405557 TI - [Adapting health services to the specific needs and utilization patterns of the new Spaniards. 2008 SESPAS Report]. AB - Because of the progressive increase in the number of immigrants and the uncertainty about the capacity of the Spanish health service to deal with the quantitative and qualitative increases in demand, the possibility of introducing changes to adapt our services to the new situation should be considered. Beginning with an analysis of the factors that influence health status and use of the health service, based on the National Health Survey (NHS), the European Statistics on Income and Living Conditions (EU-SILC) and the European Community Household Panel (ECHP), we compare the health profiles and patterns of medical resources utilization between the national and foreign populations. The pattern of demand for health services in the immigrant population corresponds basically to the needs of a young population in good health. According to NHS data, resource utilization among immigrants can even be lower than that among the national population. Assessing the link between health status and demand for healthcare from a dynamic point of view, by identifying variations in patterns of health and patterns of demand for healthcare, is important to identify imbalances in resources and to establish an appropriate hierarchy of preventive and treatment priorities. PMID- 18405558 TI - [Toward a new organization of public health services in Spain. 2008 SESPAS Report]. AB - Public health in Spain shows significant weaknesses. Spanish public health services respond reasonably well in crisis situations but tend to be invisible and occupy a marginal position in political agendas and in relation to health services. The organization of the public health subsystem is clearly out of date in terms of its ability to promote and protect community health, to prevent diseases, and to cope effectively with the new public health threats and challenges related to the physical and social environment in today's globalized world. Consequently, there is broad consensus on the need to rethink functions, strategies and the organization of public health in Spain, in line with European and international trends. Thus, public health reform is currently a pending challenge and a strategic priority. Indeed, some Autonomous Communities have initiated a process of modernization and change. Empowerment of public health in the political agendas and in relation to the health services is strongly recommended by promoting intersectorial approaches, the Health in All Policies strategy and Health Impact Assessment. There is also a need for a specific law that would update public health functions, organization and structures, allocate competencies by facilitating alliances and partnership, and regulate coordination and intersectorial intervention. The following key elements related to this reform are described: 1) a participatory leadership in public health; 2) the generation of intelligence and evidence in public health; 3) improvement of professional education and development; 4) the importance of transparent, independent and competent performance and communication, and 5) new and flexible organization coherent with the new strategies and close to the local level and primary health care services. Coordination between the State and the Autonomous Communities should involved a functional and intelligent relationship by building up common spaces, alliances, networks and shared initiatives for public health. PMID- 18405559 TI - [Health policies (population interventions) in health services. 2008 SESPAS Report]. AB - Health interventions addressed to the population as a whole from health care services are scarce and only exceptionally involve coordination among public health services (vaccinations and, in some autonomous communities, secondary breast cancer prevention). Health education programs addressed to schools are one of the most frequent interventions but their outcomes are not systematically evaluated. However, primary health care services carry out many clinical preventive activities. While the aims of these activities are laudable, the interventions themselves have substantial limitations, because they are an important source of dependency, a powerful incentive to consume drugs, and are also inefficient and inequitable ways of spending health resources. These limitations justify the testing of combined approaches between public health services and citizens' collectives to improve and protect community health. Developing community health programs based on cooperation between primary health care services and public health services requires strategies that produce appreciable results in the short term to both health sectors, as well as to the population, so that these programs stimulate the process and encourage further development. The settings in which collaboration is most promising are population health surveillance and monitoring in basic health areas, control of communicable diseases and epidemic outbreaks, health promotion and health protection programs through simultaneous clinical and community-based interventions, and improved management of all health services in local communities through joint evaluation. The resources needed to carry out these activities should be drawn from a reduction of clinical preventive activities that reduce workload and from an increase in the number and quality of the public health workforce. PMID- 18405560 TI - [The impact of drugs on social welfare. 2008 SESPAS Report]. AB - The reasons and rationale for state intervention in the pharmaceutical industry are discussed with emphasis on two of the most frequent perspectives: the health focused view and the income-centered approach; these perspectives tend to be ignored, to come into conflict, or to be confused in Spain. The growing impact of drugs on health and on the productivity of a knowledge-intensive industry - such as the pharmaceutical sector - is discussed. The impact of drugs on welfare will depend on how prescription is dealt with within the framework of some macro policies that could clearly be improved. The present article focuses on the effectiveness of Spanish pharmaceutical policy and assesses three families of policies. Several recommendations are made: greater use of price competition, regulatory signals rewarding the most innovative drugs (those that offer the greater incremental cost-effectiveness with respect to the already existing alternatives), a shake-up of the national system of innovation, and the indispensable alignment of prescriber incentives and the health system. An integrated pharmaceutical policy would help to bridge the quality chasm of the Spanish health system by making it more desirable, and therefore sustainable. PMID- 18405561 TI - [Governability of the Spanish national health system: improving the balance between the benefits and costs of decentralization. 2008 SESPAS Report]. AB - The Spanish National Health System (SNHS) is currently in a complex equilibrium which, even if acceptable in the short term, faces major sustainability and governance challenges; the so far inconclusive outcome of the SNHS decentralization process clearly underlines the importance of seeking new alternatives to effectively bring about the real benefits of decentralization, while taking advantage of the system's economies of scale and achieving its equity objectives. To this end, four lines of action are proposed in this paper: first, redefine the SNHS based on universal public coverage, as a civic right and equal for all; second, reframe the financing system and the role of the Cohesion Fund; third, review the role of central government and its Ministry of Health, and fourth, create of a SNHS Agency headed by a Commissioner, for providing an organizational framework and identity for the SNHS, and serving as the cooperative body of the seventeen Autonomous Communities' Health Services. PMID- 18405562 TI - [How should health and healthcare priorities be set and evaluated? Prioritization methods and regional disparities. 2008 SESPAS Report]. AB - The conflict between scarce resources and unlimited needs is perhaps more prominent in the healthcare sector than in any other areas. Thus, setting priorities in health care emerges as an unavoidable task. The laudable aim of adopting any health technology that improves the population's health is impossible when confronted by budgetary constraints. Therefore, the outstanding health problems of a society and the most efficient health technologies in terms of their cost-effectiveness must be identified and patients must be prioritized, bearing in mind aspects of equity and efficiency. The present article reviews the issue of setting health care priorities by examining the experiences that have been put into practice in Spain and abroad. The problem is analyzed at three levels: the "macro" level (strategic planning, identification of higher priority areas and the selection of health care interventions); the "meso" level (incorporation of cost-effectiveness analyses into clinical practice guidelines), and the "micro" level (how to design priority systems for patients on waiting lists based on clinical and social criteria). In all these levels, there is substantial heterogeneity between Spanish regional health services, the steps that need to be taken and the ground that needs to be covered. Thus, we suggest that the first steps that some regional health services have made, together with international initiatives, could serve as a reference for the definitive incorporation of new approaches in priority setting in the Spanish health system as a whole. PMID- 18405563 TI - [Economic evaluation and decision-making in health. The role of economic evaluation in the adoption and spread of health technologies. 2008 SESPAS Report]. AB - Economic evaluation of health care technologies (EEHT) has become a basic tool in the decision-making process in health. The continuous development of methods and their direct application to reality have placed EEHT as a key item on the agenda of health policy makers across countries. The present article introduces the aims of EEHT and describes the experience of several European countries where economic evaluation studies are currently used to select the health technologies to be implemented. The role played by EEHT in Spain at the present time is then discussed. Finally, pressing tasks are identified and recommendations are made for the consolidation of EEHT as a basic element in the decision-making process of health care technologies. PMID- 18405564 TI - [Possibilities and limitations of results-based management, pay-for-performance and the redesign of incentives. 2008 SESPAS Report]. AB - The concept of pay-for-performance (P4P) encompasses different strategies that aim to stimulate health care quality improvement by remunerating healthcare providers according to their performance in specific measures of efficiency or quality. Although the effectiveness of P4P in improving quality of care is largely unknown, these systems are being widely adopted in the United Kingdom, the United States and other countries, including Spain. The elements of P4P design that are most decisive for the effectiveness of these schemes are as follows: 1) who should receive the incentives, how they should be paid, what should be rewarded, the need to incorporate risk adjustments (mainly if surrogate outcomes are used as indicators) and the need to bear organizational climate and the optimal combination of financial and non-financial incentives in mind. The most important limitations to consider are the following: 1) the exclusive focus on reducing subutilization; 2) the effect on equity; 3) the "magnifying glass" effect; 4) the validity of indicators; 5) the confusion between the recommendations of clinical guidelines and quality indicators; 6) "document engineering"; 7) paternalism; 8) the negative impact on professionalism and clinicians' internal motivation, and 9) the assumption that quality problems result from imperfect individual decisions rather than from an imperfect system. PMID- 18405566 TI - [Case and disease management and improved integration of healthcare services in Spain. 2008 SESPAS Report]. AB - The boundaries between health services levels are artificial as they are answers to health organizations' problems rather than to patients' needs. However, given that these boundaries are in place, coordination between these levels should overcome the difficulties so that patients receive the services they require. Case management (patients in specific situations, such as hospital discharge) and disease management (patients with specific diseases, such as diabetes) aim to solve problems of coordination either by improving the organization of health services or by adding a new structure to the already existing structure. These forms of management are highly attractive answers to coordination problems but have no scientific basis nor is there any empirical evidence of a positive impact on patients' health or the cost of services. Case and disease management programs are vertical structures that weaken the horizontal structure in Spain, i.e. primary care. The need for case and disease management programs is an argument to push for a pro-coordination reform of primary care that gives power, responsibility and autonomy to general practitioners, so that they would be able to coordinate clinical and social services according to patients' needs. PMID- 18405565 TI - [Integration and boundaries between health and social care. 2008 SESPAS Report]. AB - We examine the opportunities to establish integrated services in Spain. The new law on services for disabled people approved by the Spanish Parliament in December 2006 and the Mental Health Plan of the Spanish Ministry of Health are used to illustrate policies that will require a high degree of integration among organizational, financial and clinical levels. In this context, some of the following questions arise: what do we know about the effectiveness of the integration of health and social services? Who should be the targets of integration and what are the structures needed for patient-centered services? Who should be responsible for and manage these services? An outline of Leutz's five criteria for integration is reviewed and the results from international experiences are discussed. We conclude that Spain is at a crossroads for the integration of services for disabled people and people with mental health problems. We recommend that a system of integrated services be organized for people with moderate or severe disability. This system should be decentralized at the levels of local health and social services, with a single budget estimated on a capitation basis. Clinical and financial responsibility should be shared between the interdisciplinary team and the case manager (nursing or social work professionals). PMID- 18405567 TI - [Lung cancer in Spain: the current situation. 2008 SESPAS Report]. AB - Lung cancer is currently the most frequent cause of death from cancer worldwide, warranting periodic analysis of the development and results of strategies aimed at reducing the incidence, mortality and morbidity related to this disease in Spain. PREVENTIVE ASPECTS: Mortality reduction among men contrasts with the increase observed in women, reflecting changes in tobacco exposure in the last few decades. Prevention programs aimed specifically at women and youngsters should be developed. In addition, the current law of smoking prevention should be amplified and vigorously applied. ASPECTS RELATED TO EARLY DIAGNOSIS: So far, there is insufficient scientific evidence to initiate a population-based lung cancer screening program. Preferential health care circuits are required to guarantee coordination among centres and levels of care and to substantially reduce current delays in the diagnosis and treatment of patients with lung cancer. TREATMENT-RELATED ASPECTS: Therapeutic decisions in all lung cancer patients should be based on teamwork and scientific evidence. Palliative care deserves the same priority as oncological treatments. Lung cancer patients should participate more fully in therapeutic decisions. Better information systems are required for clinical assessment and research on this disease. PMID- 18405568 TI - [How should patients' utilities be incorporated into clinical decisions? 2008 SESPAS Report]. AB - There are many clinical situations in which there is no "right" decision from a technical point of view. An example of this is elective surgery, in which patients' preferences are critical. One way to integrate patients' preferences within clinical practice is the application of decision analysis. According to this approach, preferences (utilities) are assessed and are then combined with physicians' knowledge. This combination of evidence and utilities leads to the so called shared decision-making (SDM) model. The overview provided in the present article indicates that: a) The SDM model, if systematically applied, could improve treatment effectiveness and patients well being; b) clinical practice, nevertheless, faces barriers in the form of time and resource constraints, limiting the application of such a model; c) discrepancies between patients' and doctors' preferences could be narrowed if patients' utilities were included in clinical practice guidelines; d) the application of this kind of analysis seems to be scarce in Spain. Moreover, information provided to patients is probably insufficient; and e) patient decision aids, even though their use is rapidly growing, are subject to certain problems. PMID- 18405569 TI - [Toward new approaches to quality. The patient as coprotagonist. 2008 SESPAS Report]. AB - In recent decades, the doctor-patient relationship has been decisively transformed, reflecting social developments in health care and the rise of individual rights in our society. This article briefly reviews how this change has been reflected in the legal and social framework, how it affects the doctor patient relationship, how health services should adapt to this new reality, which changes are predictable, or desirable, and how health services are currently responding. The model of healthcare should adjust to the new role played by patients, requiring quality control of the information accessed - available in different formats and from distinct sources - and new attitudes and skills of health professionals and patients. The development of a homogeneous information system throughout the Spanish national health service is required. This information system should address a minimum number of key indicators and outcomes. The dissemination and analysis of these indicators and outcomes is a pressing need, not only as a comparison tool, but to identify and disseminate good practices. PMID- 18405570 TI - [Listening to the voice of the patient: an imperative. 2008 SESPAS Report]. AB - To democratize health services, citizen participation should be free, informed and involve choice. To achieve this, a formal and public system of health services' evaluation is required. The present article aims to argue the need to promote the participation of patients and persons affected by disease to achieve greater democratization of health services and improve the effectiveness of healthcare. Qualitative studies and population surveys can be used to assess the extent to which greater participation is being achieved. To this end, the present article uses information extracted from distinct qualitative studies performed in Spanish patients by the team of the Josep Laporte Foundation Library since 2000. The "paradoxes" found in diagnosis allow some recommendations to be made on health policy. There is a substantial chasm between the working of the health system and citizens' perceptions, indicating the need for a communication strategy that would help the majority of citizens - and especially patients as direct users - to identify basic elements of the system. Despite state and regional legislation on patients' rights, a large proportion of patients remain unaware of these rights. This situation reveals the possible discrepancy between legislative and executive settings. The laws shaping health policy should be evaluated to consolidate democratic processes. Providing credibility and publicizing patient rights could legitimize the introduction of the need for dialogue on patients' responsibilities. PMID- 18405572 TI - [Negative effects of individual-based preventive interventions. 2008 SESPAS Report]. AB - Because preventive interventions are usually assumed to carry few risks, research on their negative effects is scarce, as revealed by a review of the Spanish literature. The studies retrieved analyze short-term effects (such as false positive results of screening tests), but do not evaluate the long-term effects of postscreening interventions (for example: preventive drugs) or even less tangible negative effects (such as medicalization). Uncritical spread of preventive medicine is occurring, with continual proposals for new screening tests (for pre-diseases and risk factors) and new vaccines. The risk-benefit ratio of this spread is, at the very least, doubtful. The pharmaceutical industry plays a key role in this process but the role of other stakeholders (health professionals and health authorities, mass media and society) should also be reviewed. The present article highlights the role of the medical profession, since, without it, definition of new diseases and pre-diseases and the creation of guidelines on the use of new preventive interventions would not be possible. The medical profession has sufficient prestige to lead any change. Before any change occurs, however, scientific societies should conduct an internal debate on their current role in the adoption of new preventive interventions, their relationships with the pharmaceutical industry and their conflicts of interest. Some medical associations are already adopting a more critical approach to the evaluation of the benefit-risk trade-off of new interventions and are analyzing the impact of their own preventive recommendations on the medicalization of life. The adverse effects of the preventive measures implemented in health services should be continually assessed and procedures for the critical evaluation of the benefit-risk-cost trade-offs of new preventive interventions should be established before making decisions on their incorporation in the health services' portfolio. Finally, in daily clinical practice, enhancing citizen participation in decisions and adopting the approach of quaternary prevention would be desirable. PMID- 18405573 TI - [Enthusiasm for diagnostic tests: health effects and their surveillance. 2008 SESPAS Report]. AB - The medicalization of life has been encouraged by an excessive interest in health as an end in itself rather than as a means for a better life. This new concept of health has stimulated the growth of the secondary market of health and, in analogy with the knowledge society, the emergence of what is known as the "health society". Among the most popular health "products" are diagnostic tests, explaining the enthusiasm for screening programs and the new genetic technologies, especially in the detection and prognosis of cancer. Because of the iatrogenic potential of diagnostic tests and the unnecessary demand caused by their excessive use, this phenomenon is a matter of public health concern. Health administrations must face this challenge with a diverse range of interventions that range from clinical management to policy formulation including the improvement of diagnostic research. The contribution of diagnostic research has so far been limited by its irregular quality. PMID- 18405571 TI - [Lights and shadows in patient safety: study and development of strategies. 2008 SESPAS Report]. AB - Study of the frequency of healthcare-related adverse events (AE) and of their effect on patients and impact on healthcare systems has markedly increased in recent years. Indepth individual analysis of the factors contributing to these events has allowed strategies to prevent or minimize the impact of AE to be developed. However, an epidemiologic perspective is needed to explore possible associations and generalize the recommendations that can be drawn from this type of analysis. Some aspects that should been considered while developing future strategies are discussed. These aspects include the importance of patient age and comorbidities on producing AE and the use of highly technological medicine, which is sometimes unnecessary. A culture of trust and confidentiality and a spirit of collaboration among organizations are indispensable to share and learn about the initiatives prompted by the study of AE and thereby improve patient safety in the national health service. PMID- 18405574 TI - [Reform, equity and efficiency of the healthcare systems in Latin America. An analysis to inform the Spanish aid. 2008 SESPAS Report]. AB - In the 1990s, international financial multilateral agencies promoted changes in the way health systems were financed and organized. Three decades later, equity and efficiency are still central problems of the health systems in developing countries. The present article focuses on the health sector reforms introduced in Latin America in order to draw policy lessons for Spanish aid. One of those reforms, the introduction of competition in health insurance management and provision and the increase of private sector participation - managed competition , was widely promoted, despite the lack of empirical evidence and the opposition from public and scientific sectors. Years after its implementation, health system financing is still inequitable and access to health services is far from universal and adequate due to the barriers imposed by insurers, among other reasons. Moreover, segmentation in healthcare provision and inefficiency persist in healthcare systems that are expensive to manage. The Spanish state, currently undergoing a process of transformation of its aid model, should focus its efforts on redressing international agencies' policies toward strengthening public health systems in the region and, at the same time, toward improving the quantity and quality of aid at country level, favoring the leadership of receiving countries. PMID- 18405575 TI - [Who benefits from health programs aimed at the poorest? Successes and failures. 2008 SESPAS Report]. AB - International aid health programs directed at the poorest play an undoubtedly constructive role in relieving acute human suffering. These programs may, however, be counterproductive and even harmful if they obscure the need to eliminate social inequities, which are the primary cause of health inequities. The present article reviews the effectiveness of two state programs that received support from international aid to (a) improve food security in rural households and vulnerable urban groups in Colombia (1996-2005) and (b) develop a local health system in the northern area of San Salvador, El Salvador (1992-1997). International aid supports successful healthcare programs, such as food programs, could generate dependency in its beneficiaries. Factors extrinsic to cooperation (political and economic factors of the context) may affect the programs and generate social conflicts. This is the case of the program to improve access to food by subsistence production in rural areas of Colombia, which failed to reach the small farmers and benefited agricultural export production owned by wealthier groups, or interventions designed to improve health among the poorest in El Salvador. PMID- 18405576 TI - [Effectiveness of official development aid and new principles and tools for health development aid. 2008 SESPAS Report]. AB - Years after its inception, the results of official development aid in terms of poverty reduction are still limited. Among the causes, especially in the health sector, are aid dispersion and its divergence from priorities and mechanisms of recipient countries' public policies. The Paris Declaration of the High Level Forum on AIDS Effectiveness (2005) fostered an agreement on the principles to be adopted to improve the effectiveness of aid by strengthening recipient countries governments' leadership of the aid and reducing aid delivery transaction costs. Aiming at these principles, the use of new instruments -programmatic instruments- and new approaches - the sector wide approach- are being promoted to increase the effectiveness of aid. Despite some weaknesses, the assessment conducted on their utilization seems to show better performance than that achieved with traditional instruments. Spain joined the Paris Declaration in March 2005 and incorporated the use of programmatic instruments and the participation in sector-wide approaches in its new Cooperation Policy, thus initiating a significant change from previous strategies that mainly encompassed project implementation and technical advice. The new strategies directed at strengthening public policy in recipient countries provide an opportunity to increase the effectiveness of aid; however, major changes are needed for their implementation. PMID- 18405577 TI - [Final summary and policy recommendations. 2008 SESPAS Report]. PMID- 18405578 TI - [The first 10 years of EUROPREV]. PMID- 18405579 TI - [The ethics of incentives]. PMID- 18405580 TI - [Utility of an information system for primary care]. AB - OBJECTIVE: To know the usefulness and applicability of an information system for basic health areas put into operation 16 years ago. DESIGN: Descriptive study of selected indicators registered in 1994 and 2004. SETTING: Basic health areas of Barcelona, Spain. PARTICIPANTS: Population living in Barcelona, Spain, during 1994 and 2004. MEASUREMENTS: The following indicators were described, and standardized taking the Barcelona 2004 population as reference: proportion of foreign population, proportion of unemployment, number of mothers less than 20 years old, AIDS incidence rate, tuberculosis incidence rate, and traffic accident, ischaemic disease, lung cancer (for men), breast cancer (for women), and total mortality rates. Results were presented for one basic health area, its corresponding district, and Barcelona. Moreover, users of the information system were interviewed about their level of knowledge, and the adequacy, usefulness, and limitations of the system. RESULTS: Disaggregated annual information allows comparing indicators between different fields and years. The information system was found useful to users to describe the level of health of the community and to design preventive interventions. CONCLUSIONS: Although some of the indicators did not show the desired level of accuracy, the information system for basic health areas showed to be useful to know the level of health of the population assigned to primary healthcare teams. PMID- 18405581 TI - [Commentary: is a population-based information system useful for primary care?]. PMID- 18405582 TI - [Usefulness of phyto-oestrogens in reduction of blood pressure. Systematic review and meta-analysis]. AB - OBJECTIVES: To determine whether treatment with phyto-oestrogens or soya protein succeeds in lowering blood pressure. DESIGN: A systematic review, evaluating all the observation studies and clinical trials, was conducted, followed by a meta analysis to evaluate blood pressure variations in patients treated with phyto oestrogens. SEARCH: The search strategy adopted used the terms "phyto oestrogens," "soya meals," "hypertension," and "blood pressure." DATA SOURCES: The data bases MEDLINE, EMBASE, Cochrane, OVID were used, without time or language restrictions. References in the relevant articles were searched for manually. STUDY SELECTION: Two independent reviewers analysed the studies found in the search. DATA EXTRACTION: The Jadad scale was used for the clinical trials and the numerical data in the text or referred to in tables were extracted. Evaluation was made of which observational and experimental articles showed a drop in blood pressure with phyto-oestrogens and which did not. In the meta analysis, data on sample size, difference in blood pressure before and after intervention and standard deviation were extracted from each study. The weighted difference of means was used with the model of randomised effects. The Review Manager v4.2.9 programme was used. RESULTS: No significant variations in blood pressure were found, whether systolic (-1.20 mm Hg; 95% CI, -2.80 to 0.41 mm Hg) or diastolic (-1.31 mm Hg; 95% CI, -2.73 to 0.11). If there were any variations, they are clinically of little importance. There was also an important degree of both statistical and clinical heterogeneity. CONCLUSIONS: There are no statistically significant or clinically important differences in blood pressure between patients treated with phyto-oestrogens and those not treated. PMID- 18405583 TI - [Opportune intervention of family doctors in the programme of early detection of breast cancer]. AB - OBJECTIVE: To study to what extent a brief intervention increases the rate of participation in the programme of early diagnosis of breast cancer (EDBC). DESIGN: Before-and-after intervention study without a control group on women who had not had a mammography. SETTING: Primary care: 3 rural lists and 2 urban ones. PARTICIPANTS: All women between 52 and 67 who attended the clinic during a 6 month period. INTERVENTION: If they had had no mammography in the previous 2 years, the reasons were explored and a brief structured intervention, depending on the reason, was conducted. After this, the woman was invited to make an appointment with the EDBC Unit. MAIN MEASUREMENTS: Effective intervention: checking that the mammography had been done at the EDBC Unit. It was checked whether women who had not attended for consultation attended the unit with their mammography done. RESULTS: Out of a target population of 565 women, 403 (71%) attended for consultation. Of these, 315 (78%) were in the programme, 51 (13%) had had a mammography outside the programme, and 37 (9%) had not had a mammography. After the intervention, 21 women had a mammography (56.7% [95% CI, 41-73] success of intervention). Seventy of the 162 women who did not attend for consultation had a mammography in the programme. The rate of participation in the EDBC rose from 68% (385/565) to 72% (406/565). CONCLUSIONS: The brief intervention was effective in over half the women. This small increase contributed to reaching the minimum levels of coverage recommended in a programme of this kind. PMID- 18405584 TI - [Overload felt by the figure of the main caregiver in a cohort of patients with multiple pathologies]. AB - OBJECTIVE: To determine the profile of the main caregiver (MC) and the factors associated with her/his care burden, in a multi-centre cohort of patients with multiple pathologies (PMP). DESIGN: Multi-centre cross-sectional study. SETTING: Four health districts in the Virgen del Rocio University Hospitals Health Area, Seville, Spain. PARTICIPANTS: The PMP cohort was created by checking all the patients who satisfied the health department criteria for PMP (2002): patients suffering from chronic diseases in 2 or more of the 7 clinical categories defined. MAIN MEASUREMENTS: The profile of PMP caregiver was determined for all patients. The caregiver strain index (CSI) was determined by the index of care stress (ICS). Predictive factors were analysed by the Student t, ANOVA, and Pearson's tests. Multivariate analysis was performed by a forward stepwise linear regression model. RESULTS: The interview was attended by 461 (69%) out of 662 eligible PMP. Of these, 293 (63.6%) had an MC whose mean age was 62 (15) years; 80% of them were women. First-degree relatives made up 88% of caregivers, with spouses 49.7% of them (n=146). In 41.5%, the CSI was >7 points (mean CSI was 5.35 [3.5]). This was higher in those caring for PMP with neurological illnesses (7 [3.2 vs 4.5 [3.3]; P=.0001). The CSI was compared directly with the medical vulnerability of the PMP (R=0.37; P=.001), cognitive deterioration on the Pfeiffer scale (PS) (R=0.4; P=.0001), and inversely with functional status on Barthel's scale (BS) (R=-0.67; P=.0001). Patient's age (P=.03), his/her medical vulnerability (P=.016) and functional (P< .0001) and cognitive (P=.019) deterioration were independently associated with the CSI. CONCLUSIONS: The profile of the MC of the PMP cohort corresponded mainly to first-degree female relatives around sixty years old. The burden of care was high in more than a third of them. Predictive factors were age, medical vulnerability, and the functional and cognitive deterioration of the PMP. PMID- 18405585 TI - [Abdominal obesity and cardiometabolic risk]. PMID- 18405586 TI - [Human papillomavirus vaccination: attitude to a consultation on a therapeutic novelty]. PMID- 18405587 TI - [Factors associated with temporomandibular disorder in a health centre's population]. PMID- 18405588 TI - [Evaluation of clinical interns for their clinical aptitude in diabetes mellitus]. PMID- 18405589 TI - [Relationship between cognitive deterioration and depression in the over-65s]. PMID- 18405590 TI - [Compliance with the metabolic control objectives in patients with type-2 diabetes attended in primary care]. PMID- 18405591 TI - [Map towards adventure]. PMID- 18405595 TI - [Clinical diagnosis of streptococcal pharyngoamygdalitis]. PMID- 18405596 TI - [Design, implementation and methodological issues in cohort study]. AB - Cohort study is graded as level II in evidence grading system of evidence-based medicine, secondary to randomized controlled trial. It is one of the important evidence for the evaluation of health care interventions, and it has been applied recently in the evaluation of traditional medicine. The author of the article introduces the basic concept, principle, design, and implementation as well as relevant methodological issues when applied in traditional Chinese medicine. PMID- 18405597 TI - [How to assess the quality of systematic review and meta-analysis]. AB - Evidence-based medicine advocates that decision making should be based on the adequate evidence. High quality systematic review is an important source of information for clinicians, patients, governments and other decision makers in decision making. If reviewers couldn't master the correct method of systematic review or meta-analysis was misused, the inaccurate, biased or misleading outcomes would be produced. So understanding of the methods of evaluating the quality of systematic review and meta-analysis is helpful to make good use of the review's information. PMID- 18405598 TI - [Evaluation of Chinese version of the Functional Assessment of Cancer Therapy Hepatobiliary questionnaire]. AB - OBJECTIVE: To evaluate the effectiveness of the Functional Assessment of Cancer Therapy-Hepatobiliary (FACT-Hep) questionnaire in measuring the quality of life in patients with primary hepatic carcinoma (PHC) in China. METHODS: FACT-Hep questionnaire was translated into Chinese and revised properly. From September 2005 to April 2006, one hundred and eighty patients with primary liver carcinoma were admitted and measured by using the Chinese version of FACT-Hep questionnaire, and the reliabilities, validities and responsibilities of the questionnaire were assessed. RESULTS: Correlation coefficient was higher between items and dimension of their corresponding domain (0.5933+/-0.1652) than that between the items and other domains (0.2749+/-0.1922). Six principal constituents were extracted by factor analysis and represented all domains of the questionnaire. The combinations of components were consistent with what was expected. The correlation coefficient of criterion-related validity was 0.828. The test-retest reliability correlation coefficients of physical, social/family, emotion, function, symptom and total questionnaire were 0.731, 0.334, 0.953, 0.786, 0.785 and 0.801 respectively, and the values of Cronbach's alpha were 0.7397, 0.4193, 0.7914, 0.8250, 0.8399 and 0.9161, respectively. There were statistical differences in scores of FACT-Hep questionnaire in different PHC stages or in different Child-Pugh classes (P<0.05). CONCLUSION: The FACT-Hep questionnaire can measure the quality of life in patients with PHC with good reliability, validity and responsiveness; it can be used in assessing the disease specific health-related quality of life of patients with hepatobiliary cancers. PMID- 18405599 TI - [Characteristics of traditional Chinese medicine syndromes in patients with acute ischemic stroke of yin or yang syndrome: a multicenter trial]. AB - OBJECTIVE: To explore the composition characteristics of traditional Chinese medicine (TCM) syndromes in patients with acute ischemic stroke of yin or yang syndrome by investigating the characteristics of TCM syndromes at different periods after onset. METHODS: One thousand two hundred and forty-six patients with acute ischemic stroke were admitted in twenty hospitals. According to the "diagnostic criteria of syndrome differentiation of stroke", the characteristics of syndromes in the patients were investigated at the periods of 1-3 days, 4-10 days and 11-30 days after they had ischemic stroke. General distribution of six basic syndromes was compared between the patients with yin syndrome and the patients with yang syndrome at the three periods. The six basic syndromes were wind syndrome, pathogenic fire syndrome, phlegm syndrome, blood stasis syndrome, qi deficiency syndrome, and syndrome of yin deficiency and yang hyperactivity. RESULTS: The percentages of wind, pathogenic fire, and phlegm syndromes in the patients were decreased at the period of 11-30 days as compared with the period of 1-3 days (87.1% vs 79.3%, 52.1% vs 38.7% and 67.1% vs 57.4% respectively, P<0.01). However, the percentages of the syndromes of blood stasis, qi deficiency, and yin deficiency and yang hyperactivity were similar at the three periods (P>0.05). There were no differences in the distribution of yin and yang syndromes among the three periods (P>0.05). The percentages of syndromes of wind, pathogenic fire, phlegm, and yin deficiency and yang hyperactivity were higher (P<0.01), and the percentages of syndromes of blood stasis and qi deficiency were lower (P<0.05, P<0.01) in patients with yang syndrome than in patients with yin syndrome. The complex of three syndromes was the most frequent composition pattern in the patients at the three periods. The percentages of complex syndromes of four or five syndromes were higher, and the percentages of single syndromes and complex syndromes of two syndromes were lower in patients with yang syndrome than in patients with yin syndrome (P<0.05, P<0.01). The most frequent complex syndromes in patients with yin syndrome were complex syndrome of wind, phlegm, blood stasis and qi deficiency, and complex syndrome of wind, phlegm and qi deficiency; while the most frequent complex syndromes in patients with yang syndrome were complex syndrome of wind, pathogenic fire, phlegm and qi deficiency, and complex syndrome of wind, pathogenic fire and phlegm. CONCLUSION: The main discrimination between the yin and yang syndromes is that the yang syndrome is characterized by pathogenic fire. The syndromes of phlegm, qi deficiency, and blood stasis are not associated with the diagnosis of yin or yang syndrome. PMID- 18405600 TI - [Risk factor analysis of severe acute pancreatitis complicated by pancreatic encephalopathy]. AB - OBJECTIVE: To study the risk factors for severe acute pancreatitis (SAP) complicated by pancreatic encephalopathy (PE). METHODS: Clinical data from 255 patients with SAP from January 2005 to December 2006 were reviewed. Thirty-one SAP patients had PE, and 224 SAP patients did not. Clinical characteristics of SAP patients in both PE group and non-PE group were analyzed. RESULTS: Ranson scale and the incidence rates of acute respiratory distress syndrome (ARDS), renal failure, hypoproteinemia, hypocalcemia and hyperglycosemia in PE group were higher than those in non-PE group (P<0.05). There were no significant differences in acute physiology and chronic health evaluation II and CT severity index scales, the activities of amylase and lipase, the incidence rate of liver function failure, the infection rate and the operability between the PE group and the non-PE group (P>0.05). Multivariate logistic regression analysis showed that ARDS and hyperglycosemia were high risk factors. Cure rate in PE group was higher than that in non-PE group. CONCLUSION: Nosogenesis of PE is the result of multiple factors. ARDS and hyperglycosemia may be the high risk factors for PE. PMID- 18405601 TI - [Investigation of gene expression profiles in patients with blood stasis syndrome]. AB - OBJECTIVE: To investigate the differential gene expression profiles in patients with blood stasis syndrome by oligonucleotide microarray technique. METHODS: Sixteen patients with blood stasis syndrome were divided into patients with coronary heart disease (CAD) (n=8) and non-CAD patients (n=8) by using coronary angiography. The sex- and age-matched eight healthy persons were enrolled as control group. Venous bloods were collected for extracting RNA. Test-3 chip was first employed to examine the quality of samples. Then the samples were hybridized with Affymetrix U133 Plus 2.0 array to compare the gene expression profiles among the three groups. Gene-array scanner and gene chip operating software were applied to screen hybridization signals and analyze gene expression respectively. Based on the comparison of the three groups of samples, the differential genes related with blood stasis syndrome were analyzed by Gene Ontology (GO) and pathway, and confirmed by real-time reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Forty-eight differential genes were found being associated with blood stasis syndrome, including 26 up-regulated genes and 22 down-regulated genes. Five of the forty-eight genes (10.4%) were related to inflammatory reaction and immune response through the GO analysis. In the pathway analysis, five of ten significant pathways were referred to inflammation and immune response. The results of real-time RT-PCR proved the accuracy of the gene chip. CONCLUSION: Inflammatory- and immune-related genes have a remarkable predominance in blood stasis syndrome gene expression profiles, which may explain the function of inflammation and immune response in the occurrence and development of blood stasis syndrome. PMID- 18405602 TI - [Effects of Bushen Huayu Qutan Recipe on local ovarian factors in androgen sterilized rats]. AB - OBJECTIVE: To explore the effects of Bushen Huayu Qutan Recipe (BHQR), a compound traditional Chinese herbal medicine, on local ovarian factors in androgen sterilized rats (ASR). METHODS: One hundred and five 9-day-old SD rats were randomly divided into normal control group (15 rats) and ASR group (90 rats). ASR was induced by subcutaneous injection of testosterone propionate, and the rat model of ASR was established successfully in 30 rats in the ASR group. Thirty ASR rats were randomly divided into untreated group and BHQR-treated group. There were 15 rats in each group. The rats in BHQR-treated group were intragastrically administered with BHQR. The rats in the normal control and untreated groups were intragastrically administered with distilled water. They were all treated for 30 days. The levels of serum testosterone and insulin (INS) were measured by radioimmunoassay. The inhibin (INH) level, and the expressions of insulin-like growth factor-I(IGF-I) and vascular endothelial growth factor (VEGF) in ovary were determined by immunohistochemical method. RESULTS: None of the ASR ovulated and all the rats in the normal control group ovulated. The rate of ovulation in BHQR-treated group was 66.67%. The serum concentrations of testosterone and INS in the untreated group were significantly higher than those in the normal control group (P<0.05), and the serum concentrations of testosterone and INS in BHQR treated group were decreased as compared with the untreated group (P<0.05). The levels of ovarian INH, IGF-I and VEGF in the untreated group were significantly higher than those in the normal control group (P<0.01), and the levels of ovarian INH, IGF-I and VEGF in BHQR-treated group were decreased as compared with the untreated group (P<0.01). CONCLUSION: BHQR is an effective recipe in promoting ovulation, and the effects of BHQR on balancing the internal environment of ovary may be due to the reduction of serum INS level and decrease in the expressions of ovarian INH, IGF-I and VEGF. PMID- 18405603 TI - [Therapeutic effects of Tiaozhong Granule on esophagus mucous membrane in rats with mixed reflux esophagitis and the mechanisms]. AB - OBJECTIVE: To explore the mechanism of Tiaozhong Granule (TZG), a compound traditional Chinese herbal medicine, in treating rats with mixed reflux esophagitis. METHODS: Fifty-eight SD rats were randomly divided into untreated group (n=12), sham-operated group (n=10), TZG-treated group (n=12), Banxia Xiexin Decoction (BXXXD)-treated group (n=12) and cisapride-treated group (n=12). Mixed reflux esophagitis was induced by esophago-duodenum end-to-side anastomosis. Four weeks later, the rats were orally administered twice daily for 12 days. Pathological changes of esophagus mucous membrane were observed by using HE staining. The expressions of proliferating cell nuclear antigen (PCNA) and p53 in the esophagus tissue were detected by immunohistochemical SABC method. Spectrophotometric method was used to detect the content of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH Px) in serum. RESULTS: Compared with the untreated group, pathological changes of esophagus mucous membrane were relieved in different degrees in TZG-treated group, BXXXD-treated group and cisapride-treated group. Content of MDA and expressions of PCNA and p53 were obviously decreased in the three treated groups (P<0.01), and the activities of SOD and GSH-Px were significantly increased in the three treated groups (P<0.05, P<0.01). TZG had better effects than cisapride in decreasing the content of MDA and increasing the activities of SOD and GSH-Px (P<0.05). TZG was better in aspect of reducing the expressions of PCNA and p53 than BXXXD and cisapride tablets (P<0.05). CONCLUSION: Tiaozhong Granule can treat mixed reflux esophagitis in rats, and its action mechanisms may be associated with decreasing the expressions of PCNA and p53 in esophagus mucous membrane, reducing the content of MDA and increasing the activities of SOD and GSH-Px in serum. PMID- 18405604 TI - [Effects of Yanghe Decoction on vascular endothelial growth factor in cartilage cells of osteoarthritis rabbits]. AB - OBJECTIVE: To observe the effects of Yanghe Decoction on vascular endothelial growth factor (VEGF) in cartilage cells of osteoarthritis rabbits. METHODS: Fifteen New Zealand white rabbits were randomly divided into normal group, untreated group and Yanghe Decoction-treated group. The rabbit model of osteoarthritis was established according to Hulth's method. The rabbits were sacrificed at the 8th week after administration of Yanghe Decoction for 14 days, and then rabbit tibia articular cartilage was removed. Sections of the cartilage were stained with Safranin O for histological examination. The cartilage histological characteristics were observed according to the method of Mankin. Immunohistochemical staining was performed to investigate the expression of VEGF. Articular cartilages were observed with microscopy and image analysis method was used to measure the expression intensity of VEGF. RESULTS: There were significant differences in Mankin score between normal group and untreated group (P<0.01), and between untreated group and Yanghe Decoction-treated group (P<0.01). Immunohistochemical staining indicated that the expression intensity of VEGF in untreated group was significantly increased as compared with that in normal group (P<0.01), and also obviously higher than Yanghe Decotion-treated group. CONCLUSION: VEGF plays an important role during early stage of OA. Yanghe Decoction can protect the articular cartilage through suppressing the VEGF expression in chondrocytes and then suppress angiogenesis. PMID- 18405605 TI - [Comparative study of Scrophulariae and Aconite in inhibiting myocardial hypertrophy in rats and mice]. AB - OBJECTIVE: To explore the effects of Scrophulariae of cold nature and Aconite of hot nature on myocardial hypertrophy and neuroendocrine factors in rats and mice. METHODS: A mouse model of myocardial hypertrophy was established by hypodermic injection of isoproterenol. Sixty myocardial hypertrophy mice were randomly divided into five groups: normal control group, untreated group, metoprolol treated group, Scrophulariae-treated group and Aconite-treated group. A rat model of myocardial hypertrophy was established by peritoneal injection of L-thyroxin. Fifty rats were randomly divided into five groups: normal control group, untreated group, captopril-treated group, Scrophulariae-treated group and Aconite treated group. After 7-9 days of treatment with intragastric administration of the corresponding drugs, the effects of Scrophulariae and Aconite on left ventricular weight index (LVWI) and heart weight index (HWI) were determined. The concentrations of cyclic adenosine monophosphate (cAMP) in plasma and angiotensin II (Ang II) in myocardium were detected through radio-immunity method. Cardiocyte cross-section area was determined by using HE staining. RESULTS: Scrophulariae of cold nature could significantly reduce the LVWI, HWI and cardiocyte cross-section area, and could decrease the content of cAMP and Ang II. However, Aconite had no such effects. CONCLUSION: Scrophulariae of cold nature can inhibit myocardial hypertrophy through restraining the activity of sympathetic nervous system and decreasing the level of Ang II. The inhibition of Aconite of hot nature on cardiac hypertrophy is not significant. PMID- 18405606 TI - [Effects of Xuefu Zhuyu Capsule on endothelin-1 release in myocardium and vascular endothelium and nitric oxide/nitric oxide synthase system of swines after acute myocardial infarction and reperfusion]. AB - OBJECTIVE: To observe the effects of Xuefu Zhuyu Capsule (XFZYC), a compound traditional Chinese herbal medicine, on endothelin-1 (ET-1) release in myocardium and vascular endothelium and nitric oxide (NO)/nitric oxide synthase (NOS) system of swines after acute myocardial infarction (AMI) and reperfusion, and to explore the action mechanisms of XFZYC in improving the endothelium function. METHODS: Forty-five Yorkshire swines were randomized into 3 groups: sham-operated group, untreated group and XFZYC-treated group. A Yorkshire swine model of reperfusion in AMI was established by ligation of left anterior descending coronary artery for 90 min followed by 2 h relaxation. The content of serum ET-1 and NO was measured by radioimmunoassay before and after AMI and after reperfusion, respectively. Twenty-four hours after operation, all Yorkshire swines underwent diagnostic coronary angiography to delineate coronary arteries. The expressions of ET-1 and endothelial nitric oxide synthase (eNOS) in myocardial tissue of ischemic area were quantified with Western blotting. Microvessel density of the implanting sites was assessed by using HE staining. RESULTS: Compared with the untreated group, the levels of serum ET-1 after AMI and reperfusion were significantly decreased in XFZYC-treated group (P<0.01), while the NO levels after AMI and reperfusion in XFZYC-treated group were significantly increased (P<0.01). There was no significant difference in diagnostic coronary angiography between XFZYC-treated group and untreated group (P=0.253). Western blotting showed that the level of ET-1 in ischemic area in XFZYC-treated group was lower than that in the untreated group (P<0.01), while the eNOS protein expression in XFZYC-treated group was higher than that in the untreated group (P<0.01). The results of HE staining and microvessel density analysis of the implanting sites all showed that the degree of telangiectasis was reduced, the cardiac muscle damage was improved, and the density of capillaries was increased obviously in XFZYC-treated group as compared with the untreated group. CONCLUSION: The endothelium injury may be one of the important mechanisms for no-reflow phenomenon. XFZYC may reduce the no-reflow by protecting endothelium cells. PMID- 18405607 TI - [Effects of Huoxue Qianyang Formula on expressions of proto-oncogenes c-fos and c myc in spontaneous hypertensive rats with ventricular hypertrophy]. AB - OBJECTIVE: To explore the possible mechanism of Huoxue Qianyang Formula (HXQYF), a compound traditional Chinese herbal medicine, in reversing the left ventricular hypertrophy (LVH) of spontaneous hypertensive rats (SHRs) by analyzing the expressions of mRNAs and proteins of proto-oncogenes c-fos and c-myc in left ventricular muscle. METHODS: The experimental study was carried out in SHRs, the sex- and age-matched Wistar-Kyoto (WKY) rats were served as normal control (n=5, normal saline 10 ml/kg daily). Twenty-five SHRs were randomly divided into five groups: untreated group (n=5, normal saline 10 ml/kg daily), high-dose HXQYF treated group (n=5, 0.84 g/ml HXQYF, 10 ml/kg daily), medium-dose HXQYF-treated group (n=5, 0.42 g/ml HXQYF, 10 ml/kg daily), low-dose HXQYF-treated group (n=5, 0.21 g/ml HXQYF, 10 ml/kg daily) and cilazapril-treated group (n=5, 1 mg/ml cilazapril, 10 ml/kg daily). The drugs were intragastrically administered once daily for 14 weeks. The expressions of mRNAs and proteins of proto-oncogenes c fos and c-myc in left ventricular muscle were detected separately by in situ hybridization histochemical method and immunohistochemical method. RESULTS: Compared with the normal control group, the expressions of mRNAs and proteins of proto-oncogenes c-fos and c-myc in left ventricular muscle were significantly increased in untreated group (P<0.01). After treatment, the expressions of c-fos and c-myc mRNAs in left ventricular muscle in HXQYF-treated groups were significantly down-regulated as compared with those of the untreated group (P<0.05). The expressions of c-myc protein were also significantly decreased in high- and medium-dose HXQYF-treated groups as compared with the untreated group (P<0.05), but it had no significant effects in protein expression of c-fos in the three HXQYF-treated groups. CONCLUSION: HXQYF can inhibit the expression of c-myc in ventricular hypertrophy tissue, which may be the mechanism in treating LVH of hypertension. PMID- 18405608 TI - [Absorption and transport of 6 coumarins isolated from the roots of Angelica pubescens f. biserrata in human Caco-2 cell monolayer model]. AB - OBJECTIVE: To study the absorption and transepithelial transport of six coumarins (umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B, isolated from the roots of Angelica pubescens f. biserrata) in the human Caco-2 cell monolayer model. METHODS: The in vitro cultured human colon carcinoma cell line, Caco-2 cell monolayer model, was applied to study the absorption and transport of the six coumarins from apical (AP) to basolateral (BL) side and from BL to AP side. The six coumarins were measured by reversed phase high-performance liquid chromatography (HPLC) coupled with ultraviolet absorption detector. Transport parameters and apparent permeability coefficients (P(app)) were calculated and compared with those of propranolol as a control substance of high permeability and atenolol as a control substance of poor permeability. The transport mechanism of angelol-B was assayed by using iodoacetamide as a reference standard to inhibit ATP-dependent transport and MK571 as a well-known inhibitor of MRP2. RESULTS: The absorption and transport of six coumarins were passive diffusion as the dominating process. The P(app) values of umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B from AP to BL side were (2.679+/-0.263) x 10(-5), (1.306+/-0.324) x 10( 5), (0.595+/-0.086) x 10(-6), (2.930+/-0.410) x 10(-6), (1.532+/-0.444) x 10(-5) and (1.413+/-0.243) x 10(-5) cm/s, and from BL to AP side were (3.381+/-0.410) x 10(-5), (0.898+/-0.134) x 10(-5), (0.510+/-0.183) x 10(-6), (0.222+/-0.025) x 10( 6), (1.203+/-0.280) x 10(-5) and (0.754+/-0.092) x 10(-5) cm/s, respectively. In this assay, the P(app) value of propranolol was 2.18 x 10(-5) cm/s and the P(app) value of atenolol was 2.77 x 10(-7) cm/s. Among the 6 coumarins, the P(app) values of umbelliferone, osthole, angelol-A and angelol-B from AP to BL side were identical with that of propranolol, and columbianadin and columbianetin acetate lied between propranolol and atenolol. When replaced the HBSS with EBSS, and iodoacetamide or MK-591 were used in the experiment, the P(app) of angelol-B had no statistical difference as compared with the control group. In the mean total recoveries, umbelliferone was (83.31+/-3.52)%, angelol-A was (77.39+/-7.38)%, osthole, columbianadin and angelol-B were between 50% to 65%, and columbianetin acetate was lower than 10%. The accumulation rates of osthole and columbianadin in the Caco-2 cells were (36.15+/-5.87)% and (53.90+/-4.39)%, respectively. CONCLUSION: The absorption and transport of umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B are passive diffusion as the dominating process in Caco-2 cell monolayer model. Umbelliferone, osthole, angelol-A and angelol-B are estimated to be highly absorbed compounds, and columbianadin and columbianetin acetate are estimated to be moderately absorbed compounds. In the Caco-2 cells, osthol and columbianadin appear to accumulate, and columbianetin acetate may be metabolized. The absorption and transport of angelol-B are not influenced by the change of pH and the presence of iodoacetamide or MK571. PMID- 18405609 TI - [Effects of Astragalus injection on proliferation of basal-like breast cancer cell line MDA-MB-468]. AB - OBJECTIVE: To investigate the effects of Astragalus injection (AI) on basal-like breast cancer cell line MDA-MB-468 and murine bone marrow stromal stem cells (mMSCs). METHODS: MDA-MB-468 cells and primary cultured mMSCs were treated by different concentrations of AI, and with untreated MDA-MB-468 cells as blank control. The morphology of cells was observed by phase-contrast inverted microscope and transmission electron microscopy. Cytotoxic effects of AI on MDA MB-468 cells and mMSCs were evaluated by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis of MDA-MB-468 cells induced by AI were measured by flow cytometry. Lactate dehydrogenase (LDH) activity in supernatants was measured by enzymatic colorimetric method. The expressions of epidermal growth factor receptor (EGFR) and p53 protein in MDA-MB-468 cells were evaluated by streptavidin-biotin-peroxidase complex method. RESULTS: A time-dependent cytotoxic effect of 1 g/ml AI was observed in MDA-MB-468 cells. 1 g/ml AI also had cytotoxic effect on mMSCs, but its effect was not better than cisplatin. 0.1 g/ml AI could promote the proliferation of mMSCs. Different concentrations of AI could all induce the apoptosis of MDA-MB-468 cells. There was no significant difference in LDH activity in the supernatants between blank control group and AI treated and cisplatin-treated groups. AI could down-regulate the expressions of EGFR and p53 protein. CONCLUSION: The effects of AI on MDA-MB-468 cells and mMSCs are related to the concentration of AI, and its mechanism of inhibiting the proliferation of MDA-MB-468 cells may be due to down-regulation of the expressions of EGFR and p53 protein. PMID- 18405610 TI - [Effects of Yanggan Lidan Granule on rate of gallstone formation and content of plasma cholecystokinin in guinea pigs with induced cholesterol gallstones]. AB - OBJECTIVE: To explore the effects of Yanggan Lidan Granule (YGLDG), a compound traditional Chinese herbal medicine for nourishing liver and improving choleresis, on the rate of gallstone formation and content of plasma cholecystokinin in guinea pigs with induced cholesterol gallstones. METHODS: Eighty guinea pigs were randomly divided into 4 groups, which were normal control group, untreated group, YGLDG-treated group and ursodeoxycholic acid (UDCA) treated group (n=20). Except the normal control group, gallstones were induced by high-cholesterol diet in the guinea pigs. YGLDG (1.81 g/kg daily) and UDCA (30.12 mg/kg daily) were given orally to guinea pigs in the corresponding group respectively for seven weeks; however, the guinea pigs of normal control group and untreated group were administered with normal saline. The physical signs of the guinea pigs and the rates of gallstone formation were examined, and the content of cholecystokinin (CCK) in the plasma was detected by radio-immunoassay. RESULTS: YGLDG could obviously improve the ethological signs of the guinea pigs. Gallstone formation rate of the untreated group (82.35%) was significantly increased as compared with that of the normal control group (5.26%) (P<0.01), while the gallstone formation rates of the YGLDG-treated group (27.28%) and UDCA treated group (38.89%) were lower than that of the untreated group (P<0.05). The content of CCK in plasma of the untreated group was significantly lower than that of the normal control group (P<0.01), while CCK content of the YGLDG-treated group and UDCA-treated group was lower than that of the normal control group, but both of them were significantly higher than that of the untreated group (P<0.01). The difference of CCK levels between YGLDG-treated group and UDCA-treated group had no statistical significance (P>0.05). CONCLUSION: YGLDG can significantly decrease the rate of gallstone formation in guinea pigs. It may be related to elevating the content of CCK in the plasma. PMID- 18405611 TI - [Duplication of an animal model of myocardial ischemia with blood stasis syndrome in mini-swines]. AB - OBJECTIVE: To establish an animal model of myocardial ischemia with blood stasis syndrome in mini-swines. METHODS: An animal model of myocardial ischemia was established in mini-swines by oppressing the coronary artery through the expansion of inner layer of Ameroid constrictor and the Ameroid constrictor was implanted into the distal end of the initial part of the first branch of interventricular septum of ramus descendens anterior arteriae coronariae sinistrae. Dynamic observation of behavior changes, general health status and changes of hemorheological parameters in the mini-swines were made after operation. RESULTS: The coronary angiography showed that the stenosis rate in ischemic group was more than 75% four weeks after operation. Compared with before operation and sham-operated group, there were great changes of behavior, general health status, tongue color and hemorheological parameters in ischemic group (P<0.05). CONCLUSION: The animal model of myocardial ischemia with blood stasis syndrome in mini-swines was established successfully 4 weeks after operation. The pathological process in the animal model is similar to that in the patients with chronic myocardial ischemia. So this model can be adopted in the research of myocardial ischemia with blood stasis syndrome. PMID- 18405612 TI - [Clinical observation of incision and drainage with ultracision-harmonic scalpel and thread-drawing in children with anal fistula: a report of 21 cases]. PMID- 18405613 TI - [Research of depression in traditional Chinese medicine: review and prospect]. PMID- 18405615 TI - [How to process quantitative data with design of two factors with inestimable interaction correctly]. PMID- 18405614 TI - [Transcatheter arterial chemoembolization combined with other therapies for the treatment of primary hepatic carcinoma: a review]. PMID- 18405616 TI - [Discussion on English translation of commonly used sentences in traditional Chinese medicine: part four]. PMID- 18405617 TI - [Necessity of both "domestication" version and "alienation" version during the process of English translation of traditional Chinese medicine]. PMID- 18405618 TI - [Stem cells, time for scale-up]. PMID- 18405620 TI - [Raising HDL-cholesterol: the future drug therapy for atherosclerosis?]. PMID- 18405622 TI - [The skin dermis host a new population of dendritic cells]. PMID- 18405624 TI - [Unraveling the mRNA localization code]. PMID- 18405628 TI - [Multipotent stromal cells: controversial impact on tumor development and metastasis]. PMID- 18405626 TI - [Haemodynamic induces aortic arch asymmetry]. PMID- 18405629 TI - [CD160 binding to HVEM triggers an inhibitory signal to the activated T cell]. PMID- 18405631 TI - [Mitochondria HSP90: the target to inactivate in cancer therapy?]. PMID- 18405632 TI - [Macular degeneration: retinal microglial cells are guilty]. PMID- 18405633 TI - [Striatal proteomic analysis suggests that first L-dopa dose equates to chronic exposure]. PMID- 18405634 TI - [A new mode of reglulation of FoxO1 by O-GlcNAc glycosylation: involvement in the glucotoxicity phenomenon]. PMID- 18405635 TI - [Circadian control of muscle transcriptome]. PMID- 18405636 TI - [Microalgae as cell factories producing recombinant commercial proteins]. AB - Extraction of natural substances and chemical synthesis are the main sources of pharmaceutical molecules. When possible, one may transfer the gene of the molecule in living cells creating individual factories producing on demand and in a safe way the requested molecule. Today, bacteria, yeast, mammalian cells and plants constitute the main plate-forms for various commercial products. Microalgae present numerous advantages and could offer a powerful tool for the production of commercial molecules in a near future. PMID- 18405637 TI - [Targeting telomeres to enforce cancer cells to senesce]. AB - The telomeres protect the end of chromosomes from being recognized and processed as an accidental double stranded break. In human somatic cells, telomeres shorten progressively with every round of DNA replication, leading to dysfunctional telomeres that trigger cellular senescence or apoptosis depending on the cell type. This telomere erosion appears to play a role in cell renewal, ageing and cancer. Two recent studies demonstrated in mouse that eroded telomeres in cancer cells blocked for apoptosis limit cancer formation by triggering senescence. These results suggest that provoking senescence may provide a way to cure cancer and point to new therapeutical strategies targeting specific telomeric functions. Nevertheless, an important question remains unanswered: does replicative senescence limit tumor formation in human? PMID- 18405638 TI - [Dyskeratosis congenita, a disease caused by defective telomere maintenance]. AB - Dyskeratosis congenita (DC), also called Zinsser-Cole-Engman syndrome, is a rare, often fatal, inherited disease described for the first time at the dermatological level by Zinsser in 1906. It is a very polymorphous disease at the clinical level, with several modes of inheritance. Several clinical symptoms of the disease can appear after a latency period. These features render DC particularly difficult to diagnose. Mutations of several genes can cause DC, four of them having been identified so far. However, for a majority of patients, the affected gene has not been found. Remarkably, all identified genes (DKC1, hTERC, hTERT, and NOP10) encode components of telomerase, all required for telomere length maintenance. DC is thus a unique clinical model for the study of the roles of telomerase and telomeres. Moreover, proteins encoded by the DKC1 and NOP10 genes are also components of so-called box H/ACA RNPs required for ribosome synthesis and pre-mRNA processing. Alterations of these processes could contribute to the symptoms of DC patients carrying mutations in DKC1 or NOP10. PMID- 18405639 TI - [Ryanodine receptor and heart disease]. AB - Calcium ions (Ca2+) play an essential role in cardiac excitation-contraction coupling. Ca2+ is stored in the sarcoplasmic reticulum (SR) and is release via SR Ca-release channels (ryanodine receptors, RyR2) to trigger contraction. RyR2 is a homotetramer comprising 4 pore-forming subunits. Each subunit is closely associated to regulatory proteins such as calstabine 2 (FKBP12.6), calmodulin, PKA, CamKII, calsequestrin and form a macromolecular complex that plays a critical role in pathological conditions. As a matter of fact, alterations of the channel activity and/or associated regulatory proteins can cause severe functional alterations resulting in arrhythmias and sudden death. Thus, RyR2 represent a novel therapeutic target and the discovery of a new pharmacological agent able to restore a normal RyR2 channel function represents a major challenge in the cardiac field. PMID- 18405641 TI - [Social inequalities in disability-free life expectancy in France]. PMID- 18405640 TI - [Glyceroneogenesis and PEPCK-C: pharmacological targets in type 2 diabetes]. AB - Obesity is a major risk factor for insulin resistance and type 2 diabetes. The link between hypertrophied adipose tissue and this pathology is thought to be non esterified fatty acids (NEFA) arising from adipocyte lipolysis. Sustained increase in plasma NEFA induces insulin resistance. In adipocytes, a significant part of lipolytic NEFA is re-esterified to triacylglycerol. Re-esterification requires glycerol-3-phosphate which, during fasting, is synthesized from lactate, pyruvate or certain amino acids in a metabolic pathway named glyceroneogenesis. The key enzyme in this pathway is the cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C). In this review, we postulate that thiazolidinediones exert their hypolipidemic and antidiabetic effects in adipose tissue at least in part through a rapid and selective induction of PEPCK-C gene transcription leading to increased PEPCK-C and glyceroneogenesis. Subsequent fatty acid re-esterification participates in the reduction in blood NEFA and insulin resistance. PMID- 18405642 TI - [FISCH-ESC, the first international symposium on human embryonic stem cell research]. PMID- 18405643 TI - [Ex vivo veritas]. PMID- 18405644 TI - [Telomerase... The real story, Elisabeth Blackburn]. PMID- 18405645 TI - [Current trends in Neuroeconomics]. PMID- 18405646 TI - [Red cell GLUT1 compensates for the lack of vitamin C synthesis in mammals]. PMID- 18405647 TI - [Ethical review in biomedical research]. AB - Through the difficulties encountered during the previous centuries in order for an animal to be recognized as a sensitive being, we saw the evolution of society's attitudes change from antiquity to our present day. Over the past twenty years animal testing has first evolved within a progressive regulatory framework reinforced by an ethical thinking which has, since 1990, led to the establishment of the ethics committees. The dialogue between these committees and researchers has led to the recovery of principles previously ignored such as the 3Rs (Replace, Reduce, Refine). This in turn has led to the application of improving experimental conditions, the progressive decrease in the number of animals used through a wise use and the replacement of animals by in vitro techniques in the very preliminary stages of research. Progress remains to be done, but the evolution of European regulations being amended, the formalization in France of ethics committees and the establishment of the National Ethics Committee should further contribute to the improvement of animal welfare in experimental research. PMID- 18405648 TI - Solitary hemangioma of the small bowel disclosed by wireless capsule endoscopy. AB - A nine-year-old child presented with melena and anemia. She had similar symptoms five months earlier and had undergone an extensive workup with upper gastrointestinal endoscopy and colonoscopy, both normal and 99m-Tc-RBC scintigraphy which was positive in the right lower quadrant. This time, capsule endoscopy was performed and disclosed an hemangioma with a dark spot suggesting recent bleeding in the ileum. The lesion was resected. Pathological examination revealed a transmural cavernous hemangioma. Small bowel hemangioma is a rare disease. Its diagnosis is extremely difficult and is usually obtained during surgery. Capsule endoscopy is an endoscopic technique that can improve preoperative diagnosis, as reported in the present case. PMID- 18405649 TI - [Treatment of thoracic anastomotic leaks after oesophagectomy with self-expanding and extractible covered stent]. AB - We report a case of a thoracic anastomotic leak after oesophagectomy for cancer treated by surgical debridement, drainage and an endoscopically placed self expanding stent. Intrathoracic, covered oesophageal stents appears to reduce leak morbidity after oesophagectomy and may be considered as a cost-effective treatment alternative. PMID- 18405650 TI - [Slowly regressing acute pandysautonomia associated with esophageal achalasia: a case report]. AB - Acute pandysautonomia is a rare acute autonomic neuropathy that mainly affects young women. We report a case of idiopathic acute pandysautonomia associated with an esophageal achalasia in a 30-year-old woman. The clinical features were inaugural dysphagia followed by signs of parasympathetic failure of the entire digestive tract, bladder and pupils. Twenty-four hours of electrocardiographic recording showed involvement of sympathetic adrenergic nerves. Esophageal achalasia was patent on esophageal manometry. Upper digestive tract motility was first involved and then extended to the entire digestive tract with intestinal obstruction, which required emergency ileostomy. Recovery of autonomic functions was slow. After 16 months, dysphagia and gut paresis improved and digestive continuity was restored. In case of subacute intestinal pseudo-obstruction associated with autonomic dysfunction, acute pandysautonomia should be suspected. In our report, the association with esophageal achalasia is uncommon. PMID- 18405651 TI - [Fissure syndrome of a gastrointestinal artery pseudoaneurysm in contact with a pseudocyst of the pancreas: rare, but serious complication of chronic pancreatitis]. AB - Most pseudoaneurysms (PsA) of the peripancreatic arteries cause direct erosion of the arterial wall from pancreatic enzymes that are usually in contact with or in a pseudocyst (PC). Rupturing is a rare and serious complication (90% mortality if untreated). We report the case of a 56-year-old patient with chronic alcoholic pancreatitis who developed a cephaloisthmic PC, complicated with a PsA of the gastroduodenal artery revealed by pain and deglobulization associated with cholestasis. After a diagnostic scan, emergency selective arteriography with coil embolization was performed. Five days later, hemorrhage recurred and a cephalic duodenopancreatectomy was performed. PsA of the gastroduodenal artery occur in the first 10 years of chronic pancreatitis. They are revealed by abdominal pains and/or gastrointestinal hemorrhage or shock from rupture. A scan with arterial reconstruction provides diagnosis. Arteriography is the most sensitive technique to locate the aneurysm and its branches and to perform selective embolization with coils. The failure rate is between 0 and 23%. Surgical treatment (elective ligation of the artery or partial pancreatic excision) should be limited to when embolisation fails and/or recurrent hemorrhage. PMID- 18405652 TI - [Pseudoaneurysm of the hepatic artery: rare complication of chronic pancreatitis]. AB - Bleeding from a pseudoaneurysm is a rare complication of chronic pancreatitis. We present two cases of ruptured pseudoaneurysms of the hepatic arteries. The first case involved a pancreatic pseudocyst that ruptured in the duodenum and the second resulted in an intrahepatic hematoma that compressed the bile tract causing secondary hemobilia. Angiographic embolization was the primary treatment in both cases, with surgery for the first patient and later radiological drainage in the second. The hemorrhage was controlled in both cases despite the severe prognosis and high mortality in these cases. PMID- 18405653 TI - Pancreatocolonic fistula complicating noninvasive intraductal papillary mucinous tumor of the pancreas. AB - Intraductal papillary mucinous tumors (IPMT) of the pancreas are a distinct clinicopathological entity that is increasingly recognized and whose natural history and clinical presentation are now better understood. Nevertheless, only rare cases of pancreatobiliary or pancreatodigestive fistulas complicating IPMT have been described so far and their clinicopathological significance and association with cancer remain controversial. We report a case of pancreatocolonic fistula complicating a noninvasive IPMT, and review the published literature. Unlike previous reports, IPMT complicated by fistula in nearby organs does not seem to be more often associated with invasive carcinoma: frequency is comparable in resected IPMT with or without internal fistula. Since fistulas are not a reliable clinicopathological predictor of invasive malignancy, en-bloc resection should not be routinely performed especially if extended resection increases the immediate risks or the long-term risks of surgery. PMID- 18405654 TI - [Gastrointestinal stromal tumors: role of computed tomography before and after treatment]. AB - Gastrointestinal stromal tumors (GIST) are the most frequent mesenchymal tumors of the gastrointestinal tract. They are generally located in the stomach or the small bowel and the potential for becoming malignant varies. Due to their expression of c-kit protein, a positive diagnosis as well as a specific targeted treatment by molecular biology (imatinib) are available. Computed tomography is the best imaging method for diagnosis, staging and follow-up of GIST. They appear as a well-defined exophytic mass with heterogeneous enhancement after intravenous injection of iodinated contrast material. They may contain areas of necrosis and/or haemorrhage and enlarged lymph nodes are exceedingly rare. Most common metastatic sites are the liver and mesentery. Large tumour size and liver or mesenteric metastases are computed tomography criteria for a high malignant potential. After treatment, decrease in tumour size, number and density of lesions and the disappearance of enhancing nodules suggest a positive response to imatinib therapy, whereas a new enhancing nodule within a mass is the usual pattern of recurrence. Positron emission tomography may be useful in specific cases when clinical data are inconsistent with computed tomography criteria. PMID- 18405655 TI - [Primary hydatid cyst of the pancreas: Diagnosis and surgical procedures. Report of three cases]. AB - The pancreas is an uncommon site of a hydatid cyst, even in countries where echinococcal disease is endemic. We report three cases of primary hydatid cysts of the pancreas revealed by abdominal pain. The diagnosis was based on ultrasound and CT-scan, which showed a cystic mass in the tail of the pancreas in the two first cases. In the third case, the diagnosis was determined preoperatively and a pancreatic fistula was found in the head of the pancreas during peroperative cholangiography. The echinococcal immunological test (Elisa) was positive in all cases. Surgical treatment included resection of the prominent lump in two patients and a cystogastric anastomosis in the third. Based on these three cases and a study of the literature, the difficulties of diagnosing this rare disease and the surgical procedures are discussed. PMID- 18405656 TI - Determining kinetics and affinities of protein interactions using a parallel real time label-free biosensor, the Octet. AB - ForteBio's Octet optical biosensor harnesses biolayer interferometry to detect and quantify molecular interactions using disposable fiber-optic biosensors that address samples from an open shaking microplate without any microfluidics. We recruited a monoclonal antibody against a panel of peptides to compare the Octet directly with Biacore's well-established 3000 platform and Bio-Rad's recently launched ProteOn XPR36 array system, which use surface plasmon resonance (SPR) to detect the binding of one analyte over four surfaces and six analytes over six surfaces, respectively. A sink method was used to prevent analyte from rebinding the ligand-coated Octet tips and enabled us to extract accurate kinetic rate constants, as judged by their close agreement with those determined by SPR. Although the Octet is not sensitive enough to detect the binding of small molecules directly, it can access their affinities indirectly via solution competition experiments. We conducted similar experiments on the SPR instruments to validate these measurements. The Octet is emerging as a versatile complement to other more sophisticated biosensors, and the ProteOn provides high-quality data near the sensitivity of Biacore but in a more multiplexed format. Our results provide a benchmark for assessing the performance of the above-mentioned sensors. PMID- 18405657 TI - A high-throughput pH indicator assay for screening glycosyltransferase saturation mutagenesis libraries. AB - Protein engineering using directed evolution or saturation mutagenesis at hot spots is often used to improve enzyme properties such as their substrate selectivity or stability. This requires access to robust high-throughput assays to facilitate the analysis of enzyme libraries. However, relatively few studies on directed evolution or saturation mutagenesis of glycosyltransferases have been reported in part due to a lack of suitable screening methods. In the present study we report a general screening assay for glycosyltransferases that has been developed using the blood group alpha-(1-->3)-galactosyltransferase (GTB) as a model. GTB utilizes UDP-Gal as a donor substrate and alpha-L-Fucp-(1-->2)-beta-D Galp-O-R (H antigen) as an acceptor substrate and synthesizes the blood group B antigen alpha-D-Galp-(1-->3)-[alpha-L-Fucp-(1-->2)]-beta-D-Galp-O-R. A closely related alpha-(1-->3)-N-acetylgalactosaminyltransferase (GTA) uses UDP-GalNAc as a donor with the same H acceptor, yielding the A antigen alpha-D-Galp-NAc-(1-->3) [alpha-L-Fuc(1-->2)]-beta-D-Gal-O-R. GTA and GTB are highly homologous enzymes differing in only 4 of 354 amino acids, Arg/Gly-176, Gly/Ser-235, Leu/Met-266, and Gly/Ala-268. The screening assay is based on the color change of the pH indicator bromothymol blue when a proton is released during the transfer of Gal/GalNAc from UDP-Gal/UDP-GalNAc to the acceptor substrate. Saturation mutagenesis of GTB enzyme at M214, a hot spot adjacent to the (211)DVD(213) metal binding motif, was performed and the resulting library was screened for increases in UDP-GalNAc transfer activity. Two novel mutants, M214G and M214S, identified by pH indicator screening, were purified and kinetically characterized. M214S and M214G both exhibited two-fold higher k(cat) and specific activity than wild-type GTB for UDP-GalNAc. The results confirm the importance of residue M214 for donor enzyme specificity. PMID- 18405658 TI - Molecular dynamics simulation of structural changes of lipid bilayers induced by shock waves: Effects of incident angles. AB - Unsteady and nonequilibrium molecular dynamics simulations of the response of dipalmitoylphosphatidylcholine (DPPC) bilayers to the shock waves of various incident angles are presented. The action of an incident shock wave is modeled by adding a momentum in an oblique direction to water molecules adjacent to a bilayer. We thereby elucidate the effects of incident shock angles on (i) collapse and rebound of the bilayer, (ii) lateral displacement of headgroups, (iii) tilts of lipid molecules, (iv) water penetration into the hydrophobic region of the bilayer, and (v) momentum transfer across the bilayer. The number of water molecules delivered into the hydrophobic region is found to be insensitive to incident shock angles. The most important structural changes are the lateral displacement of headgroups and tilts of lipid molecules, which are observed only in the half of the bilayer directly exposed to a shock wave for all incident shock angles studied here. As a result, only the normal component of the added oblique momentum is substantially transferred across the bilayer. This also suggests that the irradiation by shock waves may induce a jet-like streaming of the cytoplasm toward the nucleus. PMID- 18405659 TI - N-linked glycosylation and its impact on the electrophoretic mobility and function of the human proton-coupled folate transporter (HsPCFT). AB - The human proton-coupled folate transporter (HsPCFT, SLC46A1) mediates intestinal absorption of folates and transport of folates into the liver, brain and other tissues. On Western blot, HsPCFT migrates as a broad band (~55 kDa), higher than predicted (~50 kDa) in cell lines. Western blot analysis required that membrane preparations not be incubated in the loading buffer above 50 degrees C to avoid aggregation of the protein. Treatment of membrane fractions from HsPCFT transfected HeLa cells with peptidyl N-glycanase F, or cells with tunicamycin, resulted in conversion to a ~35 kDa species. Substitution of asparagine residues of two canonical glycosylation sites to glutamine, individually, yielded a ~47 kDa protein; substitution of both sites gave a smaller (~35 kDa) protein. Single mutants retained full transport activity; the double mutant retained a majority of activity. Transport function and molecular size were unchanged when the double mutant was hemagglutinin (HA) tagged at either the NH(2) or COOH terminus and probed with an anti-HA antibody excluding degradation of the deglycosylated protein. Wild-type or deglycosylated HsPCFT HA, tagged at amino or carboxyl termini, could only be visualized on the plasma membrane when HeLa cells were first permeabilized, consistent with the intracellular location of these domains. PMID- 18405660 TI - Loss of 3-chlorotyrosine by inflammatory oxidants: implications for the use of 3 chlorotyrosine as a bio-marker in vivo. AB - Activated neutrophils generate the potent oxidant hypochlorous acid (HOCl) from the enzyme myeloperoxidase (MPO). A proposed bio-marker for MPO-derived HOCl in vivo is 3-chlorotyrosine, elevated levels of which have been measured in several human inflammatory pathologies. However, it is unlikely that HOCl is produced as the sole oxidant at sites of chronic inflammation as other reactive species are also produced during the inflammatory response. The work presented shows that free and protein bound 3-chlorotyrosine is lost upon addition of the pro inflammatory oxidants, HOCl, peroxynitrite, and acidified nitrite. Furthermore, incubation of 3-chlorotyrosine with activated RAW264.7 macrophages or neutrophil like HL-60 cells resulted in significant loss of 3-chlorotyrosine. Therefore, at sites of chronic inflammation where there is concomitant ONOO(-) and HOCl formation, it is possible measurement of 3-chlorotyrosine may represent an underestimate of the true extent of tyrosine chlorination. This finding could account for some of the discrepancies reported between 3-chlorotyrosine levels in tissues in the literature. PMID- 18405661 TI - Alzheimer amyloid-beta peptides block the activation of C/EBPbeta and C/EBPdelta in glial cells. AB - Members of the CCAAT/enhancer binding protein (C/EBP) family of transcription factors have been reported to be up-regulated in Alzheimer's disease. In the present study, we have investigated the effects of amyloid-beta (Abeta) peptides on C/EBPbeta and C/EBPdelta, previously shown to be induced by inflammatory stimuli in glial cells. Surprisingly, electrophoretic mobility shift assay showed that both Abeta(1-42) and Abeta(25-35) blocked C/EBP activation induced by the inflammatory cytokine interleukin-1beta (IL-1beta) or lipopolysaccharide (LPS) in mixed primary glial cell cultures from rat. Abeta also blocked IL-1beta- or LPS induced C/EBP protein levels. The most prominent effects were observed on DNA binding activity and protein levels of C/EBPdelta. Our results demonstrate a dysregulation of C/EBP when glial cells are activated in the presence of Alzheimer Abeta peptides. PMID- 18405663 TI - X-ray diffraction measurement of the monolayer spontaneous curvature of dioleoylphosphatidylglycerol. AB - Phosphatidylglycerol (PG) is an anionic lipid commonly found in large proportions in the cell membranes of bacteria and plants and, to a lesser extent, in animal cells. PG plays an important role in the regulation and determination of the elastic properties of the membrane. Using small angle X-ray scattering experiments, we obtain that the monolayer spontaneous curvature of dioleoylphosphatidylglycerol (DOPG) is -1/150+/-0.021 nm(-1) when measured in 150 mM NaCl. When the experiments are carried out in 150 mM NaCl and 20mM MgCl(2), the value obtained for the monolayer spontaneous curvature is -1/8.7+/-0.037 nm( 1). These values are of importance in modelling the effects of curvature elastic stress in membrane lipid homeostasis in the bacterium Acholeplasma laidlawii [Alley, S.H., Barahona, M., Ces, O., Templer, R.H., in press. Biophysical regulation of lipid biosynthesis in the plasma membrane. Biophys. J.] and indicate that divalent cations can play a significant role in altering curvature elastic stress. PMID- 18405662 TI - TMP21 regulates Abeta production but does not affect caspase-3, p53, and neprilysin. AB - The presenilin (PS)-dependent gamma-secretase activity refers to a high molecular mass-complex including, besides PS1 or PS2, three other proteins recently identified, namely nicastrin, Aph-1, and Pen-2. This proteolytic complex has been shown to contribute to both gamma- and epsilon-cleavages of the beta-amyloid precursor protein (betaAPP), thereby generating beta-amyloid peptides (Abeta) and the APP intracellular domain (AICD), respectively. TMP21, a member of the p24 cargo protein family, was recently shown to interact with PS complexes. Interestingly, TMP21 modulates gamma-secretase-mediated Abeta production but does not regulate epsilon-secretase-derived AICD formation [F. Chen, H. Hasegawa, G. Schmitt-ulms, T. Kawarai, C. Bohm, T. Katayama, Y. Gu, N. Sanjo, M. Glista, E. Rogaeva, Y. Wakutami, R. Pardossi-Piquard, X. Ruan, A. Tandon, F. Checler, P. Marambaud, K. Hansen, D. Westaway, P. St. George-Hyslop, P. Fraser, TMP21 is a presenilin complex component that modulates gamma- but not epsilon-secretase activities, Nature 440 (2006) 1208-1212]. Here we investigate the functional incidence of the over-expression or depletion of TMP21 on both intracellular and secreted Abeta recoveries and AICD-associated phenotypes. First we confirm that TMP21 depletion yields increased levels of secreted Abeta40. However, we demonstrate that both staurosporine-stimulated caspase-3 activation, p53 and neprilysin expression and activity were not affected by TMP21 over-expression or depletion. Overall, our functional data further reinforce the view that TMP21 behaves as a regulator of gamma- but not epsilon-cleavages generated by PS dependent gamma-secretase complex. PMID- 18405664 TI - Giant unilamellar vesicle formation under physiologically relevant conditions. AB - We present an upgrade to the giant unilamellar vesicle (GUV) electroformation method allowing easy GUV production in different buffers and with various membrane compositions. Our experimental results reveal that lipid deposits obtained from aqueous liposome or proteoliposome dispersions are highly efficient for GUV electroformation. This is related to the ability of such dispersions to produce readily well-oriented membrane stacks. Furthermore, we present a protocol for GUV electroformation in various aqueous media, including electrolyte containing buffers at characteristic concentrations of biological fluids. This work unlocks historical barriers to GUV applications in scientific fields like biology, biochemistry, or biophysics where membrane composition, as well as its aqueous environment, should be adapted to biological significance. PMID- 18405665 TI - The vacuolar V1/V0-ATPase is involved in the release of the HOPS subunit Vps41 from vacuoles, vacuole fragmentation and fusion. AB - At yeast vacuoles, phosphorylation of the HOPS subunit Vps41 depends on the Yck3 kinase. In a screen for mutants that mimic the yck3Delta phenotype, in which Vps41 accumulates in vacuolar dots, we observed that mutants in the V0-part of the V0/V1-ATPase, in particular in vma16Delta, also accumulate Vps41. This accumulation is not due to a phosphorylation defect, but to reduced release of Vps41 from vma16Delta vacuoles. One reason could be a connection to vacuole fission, which is blocked in V-ATPase mutants. Vacuole fusion is not impaired between vacuoles lacking the V0-subunits Vma16 or Vma6 and wild-type vacuoles, whereas fusion between mutant vacuoles is reduced. Our data suggest a connection between vacuole biogenesis and membrane fusion. PMID- 18405666 TI - Interferon gamma is recognised by importin alpha/beta: enhanced nuclear localising and transactivation activities of an interferon gamma mimetic. AB - Interferon (IFN) gamma's ability to localise in the nucleus and function in gene activation has been known for some time, although the role of the conventional nuclear transporting importin molecules is unclear. Here, we demonstrate for the first time the direct recognition of IFNgamma and an IFNgamma mimetic peptide by IMPalpha and the IMPalpha/beta heterodimer, where the IFNgamma mimetic shows higher affinity. Significantly, this correlates well both with in vivo ability to target green fluorescent protein to the nucleus in transfected cells as determined by quantitative confocal laser scanning microscopy, as well as GAS promoter activity of a luciferase reporter. This has important implications for IFNgamma's anti-viral action, and the potential use of the IFNgamma mimetic in antiviral therapies. PMID- 18405667 TI - Extracellular transglutaminase 2 activates beta-catenin signaling in calcifying vascular smooth muscle cells. AB - Accumulation of transglutaminase 2 (TG2) is often associated with mineral deposits in vasculature. Here, we demonstrate that purified TG2 stimulated a 3 fold increase in matrix mineralization and up-regulation of osteoblastic markers in cultured primary vascular smooth muscle cells (VSMCs). Extracellular TG2 interacts with the low density lipoprotein related-protein 5 receptor and activates beta-catenin signaling in VSMCs. These results suggest that TG2 may promote vascular calcification by activating the beta-catenin signaling pathway. PMID- 18405668 TI - Enhanced ubiquitin-proteasome activity in calreticulin deficient cells: a compensatory mechanism for cell survival. AB - Calreticulin is a lectin chaperone essential for intracellular calcium homeostasis. Deletion of calreticulin gene compromises the overall quality control within the endoplasmic reticulum (ER) leading to activation of the unfolded protein response. However, the ER structure of calreticulin deficient cells (crt-/-) is not altered due to accumulation of misfolded proteins. Therefore, the aim of this study was to determine whether the ubiquitin proteasome pathway is activated in crt-/- cells as a compensatory mechanism for cell survival. Here we show a significant increase in the expression of genes involved in ER associated degradation and activation of the ubiquitin-proteasome system in crt-/- cells. We also demonstrated that the ubiquitination of two proteins processed in ER, connexin 43 and A1AT NHK (alpha1-antitrypsin mutant) are increased in crt-/- cells. Furthermore, we showed that the increased proteasome activity in the crt-/- cells could be rescued upon re-introduction of calreticulin or calsequestrin (a muscle calcium binding protein). We also illustrated that increased cytosolic Ca2+ enhances the proteasome activity. Interestingly, suppression of calnexin function using siRNA further elevated the proteasome activity in crt-/- cells. This is the first report to show that loss of calreticulin function enhances the ubiquitin-proteasome activity which could function as a compensatory mechanism for cell survival. PMID- 18405670 TI - Purification and partial characterization of canine calprotectin. AB - Calprotectin (CP) is an abundant protein in human neutrophilic granulocytes and macrophages. In humans, serum, urine, and fecal concentrations of neutrophil derived proteins, such as CP are used as markers of disease activity for conditions associated with increased neutrophil activity, such as inflammatory bowel disease. The aims of the present study were to purify and partially characterize CP in the dog (Canis familiaris) as a prelude to the development of an immunoassay for the quantification of canine serum, urine, and fecal CP in dogs with inflammatory conditions. Leukocytes were isolated from whole blood by dextran sedimentation, and canine CP (cCP) was extracted from the cytosol fraction by repeated freezing--thawing--sonication, followed by further purification using anion- and cation-exchange column chromatography. The overall yield of the purification protocol was 3.7mg cCP per 600ml whole blood. The relative molecular masses of the two proteins representing cCP (cS100A8 and cS100A9) were estimated at 10,340 and 14,628, respectively. Isoelectric focusing revealed two bands with isoelectric points of 6.4 and 6.2 for the heterodimeric protein. The approximate specific absorbance of cCP at 280nm was 0.872 for a 1mg/ml solution. The amino acid sequence of the first 13 N-terminal residues of cS100A8 was Met-Leu-Thr-Glu-Leu-Glu-Ser-Ala-Ile-Asn-Ser-Leu-Ile, whereas the N terminus of cS100A9 was blocked. Identity of both cS100A8 and cS100A9 was confirmed by tryptic peptide mass fingerprinting followed by peptide sequencing. Antibacterial activity of cCP against Escherichia coli was shown to be concentration-dependent and was reversible upon addition of micromolar amounts of zinc. We conclude that cCP can be successfully purified from canine whole blood using this reproducible, rapid and efficient method. PMID- 18405669 TI - FAK-independent alphavbeta3 integrin-EGFR complexes rescue from anoikis matrix defective fibroblasts. AB - Extracellular matrix (ECM) binding to integrin receptors regulates cell cycle progression and survival. In adherent cells, ECM disassembly induces anoikis, the apoptotic pathway switched on by loss of adhesion. ECM-deficient Ehlers-Danlos syndrome (EDS) fibroblasts, to adhere to rare fibronectin (FN) fibrils, and to proliferate, only organize, as FN receptor, the alphavbeta3 integrin. We report that in EDS cells the alphavbeta3 integrin is bound to talin and vinculin, but not to tensin, and that actin cytoskeleton is disorganized. Furthermore, in EDS cells Bcl-2 is down-regulated and caspases are active. We provide evidence that the antibody-mediated alphavbeta3 integrin or the FN inhibition induces anoikis in EDS cells. The alphavbeta3 integrin transduces survival signals to pp60src mediated tyrosine phosphorylated paxillin, instead than to FAK, and interacts with EGF receptor (EGFR). This complex, when activated by EGF and FN, signals for the rescue of EDS cells from anoikis. Therefore, EDS cells, through the alphavbeta3 integrin-EGFR complexes, engage a paxillin- but not FAK-mediated pathway of cell survival. PMID- 18405671 TI - Ethanol induces peroxynitrite-mediated toxicity through inactivation of NADP+ dependent isocitrate dehydrogenase and superoxide dismutase. AB - It has been reported that chronic alcohol administration increases peroxynitrite hepatotoxicity by enhancing concomitant production of nitric oxide and superoxide. Several studies have shown the importance of superoxide dismutase (SOD) in protecting cells against ethanol-induced oxidative stress. Recently, we demonstrated that the control of cytosolic and mitochondrial redox balance and the cellular defense against oxidative damage is one of the primary functions of NADP+-dependent isocitrate dehydrogenase (ICDH) through to supply NADPH for antioxidant systems. In this report, we demonstrate that ethanol induces the peroxynitrite-mediated cytotoxicity in HepG2 cells through inactivation of antioxidant enzymes such as ICDH and SOD. Upon exposure to 100mM ethanol for 3days to HepG2 cells, a significant decrease in the viability and activities of ICDH and SOD was observed. The ethanol-induced inactivation of antioxidant enzymes resulted in the cellular oxidative damage and modulation of redox status as well as mitochondrial dysfunction in HepG2 cells. The cytoxicity of ethanol and inactivation of antioxidant enzymes were effectively protected by manganeses(III) tetrakis(N-methyl-2-pyridyl) porphyrin, a manganese SOD mimetic, and N'-monomethyl-l-arginine, a nitric oxide synthase inhibitor. These results indicate that ethanol toxicity is mediated by peroxynitrite and the peroxynitrite mediated damage to ICDH and SOD may be resulted in the perturbation of the cellular antioxidant defense systems and subsequently lead to a pro-oxidant condition. PMID- 18405672 TI - Mass spectrometric and kinetic studies on slow progression of papain-catalyzed polymerization of L-glutamic acid diethyl ester. AB - Papain polymerizes L-glutamic acid diethyl ester (Glu-di-OEt) regioselectively, resulting in the formation of poly (gamma-ethyl alpha-L-glutamic acid) with various degrees of polymerization of less than 13. Reaction temperatures below 20 degrees C were appropriate for the reaction in terms of suppression of non enzymatic degradation of Glu-di-OEt and an increase in the peptide yield, while the reaction was preceded by a pronounced induction period. Mass spectrometric analyses of the reaction conducted at 0 degrees C revealed that the accumulation of the initial dimerization product, L-glutamyl-L-glutamic acid triethyl ester (Glu-Glu-tri-OEt), was limited during the induction period, and that a sequential polymer derived from a further elongation of the dimer was the tetramer, but not the trimer. Kinetic analyses of acyl transfer reactions with Glu-di-OEt and Glu Glu-tri-OEt as acyl acceptors and Nalpha-benzoyl-L-arginine ethyl ester as an acyl donor affirmed that Glu-Glu-tri-OEt bound more strongly than Glu-di-OEt both to the S- and S'-subsites of papain. Therefore, what occurred during the initial stage of the polymerization was interpreted as follows: the rate of the papain catalyzed dimerization of Glu-di-OEt was extremely slow, once Glu-Glu-tri-OEt was initially synthesized it exclusively bound to the active site of papain, and then papain utilized the dimer in polymerization effectively rather than the monomer. PMID- 18405674 TI - Towards a novel haemoglobin-based oxygen carrier: Euro-PEG-Hb, physico-chemical properties, vasoactivity and renal filtration. AB - Blood transfusion is still a critical therapy in many diseases, traumatic events and war battlefields. However, blood cross-matching and storage may limit its applicability, especially in Third World countries. Moreover, haemoglobin, which in red blood cells is the key player in the oxygen transport from lung to tissues, when free in the plasma causes hypertension and renal failure. This investigation was aimed at the development of a novel haemoglobin-based oxygen carrier with low vasoactivity and renal filtration properties. Human haemoglobin was chemically conjugated with polyethylene glycol (PEG) under either aerobic or anaerobic conditions, following different chemical procedures. The resulting PEGylated haemoglobin products were characterized in terms of oxygen affinity, cooperativity, effects of protons and carbon dioxide concentration, and oxidation stability, and were transfused into rats to evaluate vasoactivity and renal filtration. A deoxyhaemoglobin, conjugated with seven PEG and seven propionyl groups, which we called Euro-PEG-Hb, did not produce profound hypertension, was 99% retained within 6 h, and exhibited oxygen binding properties and allosteric effects more similar to human haemoglobin A than the other tested PEGylated haemoglobin derivatives, thus appearing a very promising candidate as blood substitute. PMID- 18405673 TI - In vivo gamma-tocopherol supplementation decreases systemic oxidative stress and cytokine responses of human monocytes in normal and asthmatic subjects. AB - We have recently reported that gamma-tocopherol (gammaT) reduces allergen- and zymosan-induced inflammation using rodent models. As an initial step in extending these observations to humans, we conducted an open-label, Phase I dosing study of two doses (one or two capsules daily for 1 week) of a gamma-tocopherol-rich preparation containing 623 mg of gamma-tocopherol, 61.1 mg of d-alpha-tocopherol, 11.1 mg of d-beta-tocopherol (11.1 mg), and 231 mg of d-sigma-tocopherol per capsule. Endpoints for this study include serum levels of 5-nitro-gamma tocopherol, as a marker of oxidative stress, and changes in serum gamma-, alpha-, and delta-tocopherol and gamma-2'-carboxyethyl-6-hydroxychroman (CEHC) 6 and 24 h after the first dose and after 1 week of treatment. To assess the biological activity of this treatment, we obtained peripheral blood mononuclear cells at baseline and after 1 week of treatment with two capsules of a gamma-tocopherol rich preparation/day and examined the inflammatory cytokine response of these cells in culture to ex vivo endotoxin/LPS (0.01 ng/ml) challenge. We also monitored a number of safety endpoints to examine how well this preparation is tolerated in eight normal volunteers (four allergic and four nonallergic) and eight allergic asthmatics. We further obtained human monocytes from a subset of these volunteers and treated them ex vivo with gammaT, alphaT, gamma-CEHC, and alpha-CEHC and assessed their actions on LPS-induced degradation of IkappaBalpha and JNK signaling and ROS generation. As detailed herein, this open-label study demonstrates that gamma-tocopherol-enriched supplementation decreased systemic oxidative stress, increased serum levels of gamma-tocopherol, and inhibited monocyte responses to LPS without any adverse health effects. Further, in vitro treatment of human monocytes with gamma-CEHC and alpha-CEHC inhibits ROS generation and LPS-induced degradation of IkappaB and JNK activation. PMID- 18405676 TI - Biochemical and structural characterization of the hypoxanthine-guanine-xanthine phosphoribosyltransferase from Pyrococcus horikoshii. AB - The 6-oxopurine phosphoribosyltransferase (HPRT, EC 2.4.2.8) from the hyperthermophile Pyrococcus horikoshii was expressed in Escherichia coli and purified. Steady-state kinetic studies indicated that the enzyme is able to use hypoxanthine, guanine and xanthine. The first two substrates showed similar catalytic efficiencies, and xanthine presented a much lower value (around 20 times lower), but the catalytic constant was comparable to that of hypoxanthine. The enzyme was not able to bind to GMP-agarose, but was able to bind the other reverse reaction substrate, inorganic pyrophosphate, with low affinity (K(d) of 4.7+/-0.1 mM). Dynamic light scattering and analytical gel filtration suggested that the enzyme exists as a homohexamer in solution. PMID- 18405675 TI - NO and CO binding profiles of hemoglobin vesicles as artificial oxygen carriers. AB - Hemoglobin vesicles (HbVs) are artificial oxygen carriers encapsulating purified and concentrated Hb solution in phospholipid vesicles (liposomes). We examined in vitro reaction profiles of a formulation of HbV with NO and CO in anaerobic and aerobic conditions using stopped-flow spectrophotometry and a NO electrode. Reaction rate constants of NO to deoxygenated and oxygenated HbV were considerably smaller than those of cell-free Hb because of the intracellular NO diffusion barrier. The reaction of CO with deoxygenated HbV was slightly slower than that of cell-free Hb solely because of the co-encapsulated allosteric effector, pyridoxal 5'-phosphate. The NO depletion in an aerobic condition in the presence of empty vesicles was monitored using a NO electrode, showing that the hydrophobic bilayer membrane of HbV, which might have higher gas solubility, does not markedly facilitate the O(2) and NO reaction, and that the intracellular Hb is the major component of NO depletion. In conclusion, HbV shows retarded gas reactions, providing some useful information to explain the absence of vasoconstriction and hypertension when they are intravenously injected. PMID- 18405677 TI - Conducting polymers in chemical sensors and arrays. AB - The review covers main applications of conducting polymers in chemical sensors and biosensors. The first part is focused on intrinsic and induced receptor properties of conducting polymers, such as pH sensitivity, sensitivity to inorganic ions and organic molecules as well as sensitivity to gases. Induced receptor properties can be also formed by molecularly imprinted polymerization or by immobilization of biological receptors. Immobilization strategies are reviewed in the second part. The third part is focused on applications of conducting polymers as transducers and includes usual optical (fluorescence, SPR, etc.) and electrical (conductometric, amperometric, potentiometric, etc.) transducing techniques as well as organic chemosensitive semiconductor devices. An assembly of stable sensing structures requires strong binding of conducting polymers to solid supports. These aspects are discussed in the next part. Finally, an application of combinatorial synthesis and high-throughput analysis to the development and optimization of sensing materials is described. PMID- 18405678 TI - Sorbent- and liquid-phase microextraction techniques and membrane-assisted extraction in combination with gas chromatographic analysis: A review. AB - Approaches are described for on-line and off-line sample pretreatment of liquid samples utilising liquid- and adsorbent- and sorbent-phase microextraction methodologies with GC analysis. Solid-phase microextraction (SPME), stir-bar sorptive extraction (SBSE), on-line solid-phase extraction (SPE), liquid-phase microextraction (LPME) and membrane-assisted methods are critically evaluated and the applicability of each technique is demonstrated with examples. PMID- 18405679 TI - Interference-free determination of thallium in aqua regia leaches from rocks, soils and sediments by D(2)-ETAAS method using mixed palladium-citric acid lithium chemical modifier. AB - The mixture of palladium (chloride) with citric acid and lithium is proposed as a new chemical modifier for the elimination of interference occurred during the determination of Tl in aqua regia extracts from rocks, soils and sediments by electrothermal atomic absorption spectrometry using instrumentation with deuterium-lamp background correction (D(2)-ETAAS). Palladium was preferred to rhodium and platinum as to analyte stabilization, citric acid served as an effective reducing agent facilitating formation of Pd-Tl stable covalent bonds playing an important role in the analyte stabilization. Citric acid in addition helps to remove most of interfering chloride at low temperature. The further addition of Li increased significantly the robustness of chemical modifier against strongly interfering ZnCl2 matrix by binding free chlorine into a more stable LiCl molecule. In the presence of the proposed chemical modifier the temperature for the final step of pyrolysis was adjustable up to 1000 degrees C, without any noticeable loss of volatile Tl species and the interference of the rest chloride matrix was significantly reduced. The application of the modifier to direct determination of Tl in aqua regia extracts from rocks, soils and sediments has ensured the characteristic mass and LOD value for the original sample 13 pg and 0.043 microg g(-1), respectively (10-microL aliquots of sample) and has enabled the use of matrix-free standard solutions for attaining accurate analysis. The accuracy was verified by the analysis of certified reference samples and by the comparison of results with those found by an inductively coupled plasma orthogonal acceleration time-of-flight mass spectrometer (ICP-oa TOFMS) method. PMID- 18405680 TI - Second-order advantage from kinetic-spectroscopic data matrices in the presence of extreme spectral overlapping A multivariate curve resolution--alternating least-squares approach. AB - Multivariate curve resolution coupled to alternating least-squares (MCR-ALS) has been employed to model kinetic-spectroscopic second-order data, with focus on the achievement of the important second-order advantage, under conditions of extreme spectral overlapping among sample components. A series of simulated examples shows that MCR-ALS can conveniently handle the studied analytical problem unlike other second-order multivariate calibration algorithms, provided matrix augmentation is implemented in the spectral mode instead of in the usual kinetic mode. The approach has also been applied to three experimental examples, which involve the determination of: (1) the antiparkinsonian carbidopa (analyte) in the presence of levodopa as a potential interferent, both reacting with cerium (IV) to produce the fluorescent species cerium (III) with different kinetics; (2) Fe(II) (analyte) in the presence of the interferent Zn(II), both catalyzing the oxidation of methyl orange with potassium bromate; and (3) tartrazine (analyte) in the presence of the interferent brilliant blue, both oxidized with potassium bromate, with the interferent leading to a product with an absorption spectrum very similar to tartrazine. The results indicate good analytical performance towards the analytes, despite the intense spectral overlapping and the presence of unexpected constituents in the test samples. PMID- 18405681 TI - Cloud point extraction and determination of trace trichlorfon by high performance liquid chromatography with ultraviolet-detection based on its catalytic effect on benzidine oxidizing. AB - 4-Amino-4'-nitrobiphenyl, which is formed by catalytic effect of trichlorfon on sodium perborate oxidizing benzidine, is extracted with a cloud point extraction method and then detected using a high performance liquid chromatography with ultraviolet detection (HPLC-UV). Under the optimum experimental conditions, there was a linear relationship between trichlorfon in the concentration range of 0.01 0.2 mgL(-1) and the peak areas of 4-amino-4'-nitrobiphenyl (r=0.996). Limit of detection was 2.0 microgL(-1), recoveries of spiked water and cabbage samples ranged between 95.4-103 and 85.2-91.2%, respectively. It was proved that the cloud point extraction (CPE) method was simple, cheap, and environment friendly than extraction with organic solvents and had more effective extraction yield. PMID- 18405682 TI - Characterization of recognition sites for diethyl 4-nitrobenzylphosphonate, an organophosphate pesticide analogue. AB - In a first step towards chemical sensors using molecular imprinted materials, the complexing characteristics of diethyl 4-nitrobenzylphosphonate, an organophosphate pesticide analogue, have been studied. Two molecules have been assessed as potential interacting moieties, specifically a fluoroalcohol and an aromatic acid. The interactions have been first characterized by regular methods, such as 1H, 31P NMR and IR spectroscopy. These showed a stoichiometry 1/1 for both complexes and association constants, respectively, close to 40+/-10 and 12+/ 2 M(-1). In a second step, isothermal titration calorimetry was used and a method was developed to obtain low-association constants. The association constant could be obtained for the fluoroalcohol ligand and was found equal to 63+/-0.7 M(-1). For the acidic molecule, an appropriate model could not be found, preventing the evaluation of this constant. PMID- 18405683 TI - Adsorption of a protein-porphyrin complex at a liquid-liquid interface studied by total internal reflection synchronous fluorescence spectroscopy. AB - Interfacial analysis has attracted more and more attention owing to its fundamental and biological importance. Total internal reflection fluorescence (TIRF) spectroscopy is a useful method to study interfacial properties. The synchronous scanning fluorescence technique provides a selective tool to analyze a specific component in a complex system. The interaction and adsorption of bovine serum albumin (BSA) and meso-tetrakis(4-sulfonatophenyl)porphyrin (TPPS) at toluene-water interface were studied successfully by the coupling technique of total internal reflection synchronous fluorescence (TIRSF). New methods are provided for the determination of the critical micelle concentration (cmc), apparent adsorption equilibrium constant (K(ad)) and maximum amount of adsorption (f(max)) at the liquid-liquid interface. The results indicated that BSA could adsorb onto the toluene-water interface as a complex of BSA-TPPS in a ratio of 1:1 ratio based on Langmuir adsorption isothermal model. The cmc, apparent K(ad) and f(max) for BSA at pH 3.1 were determined to be 1.0 x 10(-4) mol L(-1), 1.15 x 10(5) L mol(-1) and 1.14 x 10(-9) mol cm(-2), respectively. PMID- 18405684 TI - Phosphate-selective fluorescent sensing microspheres based on uranyl salophene ionophores. AB - Optical dihydrogen phosphate-selective sensors that function on the basis of bulk optode principles and are based on two different uranyl salophene ionophores are reported here for the first time. The influence of the optode composition and measuring conditions such as sample pH on the optode response are characterized, along with sensor selectivity and long-term stability. Three plasticizers of different polarity are considered for optode fabrication: bis(2 ethylhexyl)sebacate (DOS), dodecyl 2-nitrophenyl ether (o-NPDDE), o nitrophenyloctylether (o-NPOE). The compounds 9-(diethylamino)-5 (octadecanoylimino)-5H-benzo[a]phenoxazine (ETH 5294, chromoionophore I) and 9 (diethylamino)-5-[(2-octyldecyl)imino]benzo[a]phenoxazine (ETH 5350, chromoionophore III) are used as H(+)-selective fluoroionophores that also act as reference ionophores. The resulting optode-based sensors are compared with their ion-selective electrode (ISE) counterparts, and it is revealed that optodes are better suited for operation at physiological pH. The best optode performance was found for the two component optode sensors doped with ETH 5350 and phosphate ionophore(I). The linear range of these sensor was loga=-6.0 to -2.6. Dihydrogen phosphate-selective optode sensors of optimized composition are fabricated in microsphere format and preliminary measurements in diluted sheep blood samples are presented. PMID- 18405685 TI - Functional polyterthiophene-appended uranyl-salophen complex: electropolymerization and ion-selective response for monohydrogen phosphate. AB - We have synthesized a bis(terthiophene)-appended uranyl-salophen complex, comprising N,N'-bis[4-(5,2':5',2''-terthiophen-3'-yl)salicylidene]-1,2 ethanediamine-uranyl complexes (TUS), and used it as a monomer for the electrochemical polymerizations (poly-TUS) on glassy carbon surfaces to prepare functionalized conducting polymer (CP) films. The poly-TUS films prepared from propylene carbonate/0.1 M tetrabutyl ammonium perchlorate (TBAP) on a glassy carbon electrode have both the functionality of ion-to-electron transducers (solid contact) and Lewis-acidic binding sites for a monohydrogen phosphate (MHP) ion-selective electrode (ISE). The CP/poly-TUS sensor showed a linear range between 1.0 x 10(-1) and 1.0 x 10(-4.5) M with a near-Nernstian behavior (-30.4 mV decade(-1)) at a pH of 8.2. The detection limit of the electrode was 10(-5.0) M and the response time was improved (< 10s) compared to that of conventional ISEs (< 20s). For comparison, a conventional ISE (with an internal aqueous solution) based on a TUS monomer/o-nitrophenyl octylether (o-NPOE)/polyvinyl chloride (PVC) liquid membrane with or without tridodecylmethylammonium chloride (TDMACl) as an additive was also constructed and its performance as an MHP-ISE were studied. The superior selectivity and sensitivity of the CP/poly-TUS sensor enabled the direct measurement of MHP in a wide variety of applications. PMID- 18405686 TI - Disposable electrochemical DNA-array for PCR amplified detection of hazelnut allergens in foodstuffs. AB - An electrochemical low-density DNA-array has been designed and implemented to be used in combination with polymerase chain reaction (PCR) in order to investigate the presence of hazelnut major allergens (Cor a 1.04, Cor a 1.03) in foodstuff. Unmodified PCR products were captured at the sensor interface via sandwich hybridization with surface-tethered probes and biotinylated signalling probes. The resulting biotinylated hybrids were coupled with a streptavidin-alkaline phosphatase conjugate and then exposed to a alpha-naphthyl phosphate solution. Differential pulse voltammetry was finally used to detect the alpha-naphthol signal. The detection limits for Cor a 1.03 and Cor a 1.04 were 0.3 and 0.1 nmol L(-1), respectively (R.S.D. 10%). The optimized conditions were used to test several commercially available foodstuffs, claiming to contain or not the targeted nuts. The results were compared with those obtained with classical ELISA tests. PMID- 18405687 TI - Application of new sol-gel electrochemical sensors to the determination of trace mercury. AB - The aim of this work was the synthesis and characterization of new modified sol gel carbon composite electrodes and their application to the determination of trace mercury species with positive charge. Two types of modified electrodes were synthesized, sol-gel and sol-gel-PVSA carbon composite electrodes. In the last ones, poly(vinylsulfonic acid) (PVSA) was used as a functional polymer entrapped within the sol-gel material due to its cationic exchange properties. In a first stage, parameters affecting both, the sol-gel process and the electrode preparation were optimized. In a second stage, usefulness of developed electrodes applied to the determination of cationic mercury species was evaluated, optimizing the activation, preconcentration, measurement and regeneration steps. Developed electrodes showed very favourable electroanalytical properties for their use as amperometric sensors, such as small size, low cost, simple fabrication and handling, renewability and reusability. By means of an easy and low-cost methodology, satisfactory experimental results were obtained in Hg2+ determination. In this sense, developed analytical methodology showed adequate response times, linear concentration range up to three orders of magnitude (from 5.0 x 10(-8) to 5.0 x 10(-5)M) and detection limits of 1.5 x 10(-8)M (3.0 microg L(-1)). These results suggest that the incorporation of different receptor molecules at the sol-gel carbon composite material in combination with a selected electrochemical reaction could improve the detection limit achieved and obtain electrochemical sensors adapted to the determination of different species of mercury and other heavy metals. PMID- 18405688 TI - Preparation of molecularly imprinted solid phase extraction using bensulfuron methyl imprinted polymer and clean-up for the sulfonylurea-herbicides in soybean. AB - A pre-treatment methodology based on the molecularly imprinted solid phase extraction (MI-SPE) procedure was developed for the determination of bensulfuron methyl (BSM), tribenuron-methyl (TBM), metsulfuron-methyl (MSM) and nicosulfuron (NS) in soybean samples. A molecular imprinted polymer (MIP) was prepared by precipitation polymerization using BSM as the template molecule, alpha methacrylic acid (MAA) as the functional monomer, trimethylolpropane trimethacrylate (TRIM) as the cross-linker and dichloromethane as the porogen. The binding behaviors of the template BSM and its analogues on the MIP were evaluated by high performance liquid chromatography (HPLC). Then, solid phase extraction (SPE) with a BSM molecularly imprinted polymer (BSM-MIP) as adsorbent was investigated and the optimum loading, washing, and eluting conditions for MI SPE of the selected BSM, MSM, TBM, and NS were established. The optimized MI-SPE procedure was used to extract the sulfonylureas and a high recovery was obtained in the soybean samples. PMID- 18405689 TI - Incidental findings at surgery. Part 1. Foreword. PMID- 18405690 TI - Incidental findings at surgery. Part 1. In brief. PMID- 18405691 TI - Incidental findings at surgery. Part 1. PMID- 18405692 TI - Mature cystic teratoma arising at the porta hepatis: a diagnostic dilemma. AB - Mature cystic teratoma (MCT) is a tumor of embryological origin, which contains tissue derived from any or all germ cell lines found in an abnormal location. We present 2 cases of MCT both arising from the porta hepatis that were incorrectly thought perioperatively to be a choledochal cyst, which subsequently were demonstrated on histology to be a MCT. PMID- 18405693 TI - Bladder outlet obstruction causes fetal enterolithiasis in anorectal malformation with rectourinary fistula. AB - Extraluminal calcified meconium is found frequently by prenatal ultrasound in cases with bowel perforation and meconium peritonitis. Intraluminal intestinal meconium calcifications are rarely seen in prenatal sonography. Meconium calcifications result from a mixture of meconium and urine that indicates a connection between intestinal and urinary tract. We report a case of a male newborn prenatally diagnosed with intraluminal echogenic calcifications at 23 weeks of gestation, suggesting an anorectal malformation (ARM) with rectourinary fistula. At birth, the child presented with a complex ARM including high anal atresia with both perineal and rectourethral fistula. Furthermore, a bladder outlet obstruction due to a urethral stenosis was diagnosed. Vesicostomy was performed as an emergency procedure followed by colostomy during neonatal period. Posterior sagittal anorectoplasty was performed at the age of 4 months. Prenatal echogenic calcifications within bowel should raise the suspicion of ARM with rectourinary fistula and bladder outlet obstruction. PMID- 18405694 TI - Recombinant factor VIIa for intraoperative bleeding in a child with hepatoblastoma and review of recombinant activated factor VIIa use in children undergoing surgery. AB - We report a case of a child with a large liver mass who underwent an open liver biopsy and had massive bleeding intraoperatively. Recombinant activated factor VII (rFVIIa) given intraoperatively was successful in stopping the bleeding. We also reviewed the literature on the use of rFVIIa in pediatric surgery. PMID- 18405695 TI - Fibroepithelial papillomatosis ("skin tags") in Rabson-Mendenhall syndrome. AB - Rabson-Mendenhall syndrome is a rare autosomal recessive syndrome in children involving a defective insulin receptor gene. Several phenotypic features are common to this syndrome, including severe hyperinsulinemia, growth retardation, acanthosis nigricans, dental dysplasia, hirsutism, coarse facial features, and pineal hyperplasia. The authors evaluated and treated a patient with Rabson Mendenhall syndrome who presented with additional notable syndromic sequelae including extensive fibroepithelial papillomatosis ("skin tags"), not previously described to this extent. PMID- 18405696 TI - Recurrent acute thyroid swellings because of pyriform sinus fistula. AB - Anterior neck abscesses are not rare; but their origin from within the usually infection-resistant thyroid gland is not thought of in the first instance. We encountered 3 patients with differing presentations (tender nodule over anterior neck, recurrent abscess overlying the thyroid gland, and nonhealing fistula with inflammation of the anterior neck). These were caused by persistent embryological communication from the pyriform sinus to the thyroid gland to the left lobe. Excluding the first patient in whom an abnormal communication with oropharynx was suspected when actinomyces was detected in the aspiration cytology of a thyroid nodule, the other 2 patients underwent drainage as for any neck abscess. Fistulous communication was confirmed on barium swallow (in 2 patients) or computed tomographic scan (in 1 patient). En bloc excision of the affected thyroid along with the fistulous tract was performed in all patients. Long-term follow-up confirmed a cure. PMID- 18405697 TI - Conservative management of a traumatic abdominal wall hernia after a bicycle handlebar injury (case report and literature review). AB - 'Handlebar hernia' is the name given to a specific type of traumatic abdominal wall hernia (TAWH). It is an uncommon injury, caused by impact of the abdominal wall against a blunt object, often bicycle handlebars. We present a case of handelbar hernia, discuss its investigation and treatment and suggest that immediate surgical repair may not always be required. PMID- 18405698 TI - Laparoscopic aided cholecystostomy as a treatment of inspissated bile syndrome. AB - We report a case of a newborn girl with inspissated bile syndrome (IBS) that did not respond to treatment with oral ursodeoxycholic acid (Ursofalk). A solution was found using laparoscopic aided cholecystostomy with an indwelling catheter for local Ursofalk flushing in the gallbladder and the choledochus. This is the first report of a laparoscopic aided management of IBS without cholecystectomy or exploration of the bile ducts. This minimal invasive approach showed a clear advantage for the patient. There were no complications. The method is recommended in the treatment of IBS. PMID- 18405699 TI - A case of necrotizing enterocolitis associated with adenovirus infection in a term infant with 22q11 deletion syndrome. AB - Infections with adenoviruses are a common problem in the pediatric population. Normally asymptomatic to mild, those infections tend to take a more severe course in immunocompromised patients. 22q11 deletion syndrome (22q11DS) represents a common genetic disorder causing immunodeficiency from thymic hypoplasia or aplasia, heart defects, a characteristic facial appearance, and velopharyngeal dysfunction. Necrotizing enterocolitis (NEC) is a frequent gastrointestinal emergency observed in neonatal intensive care units. The occurrence of NEC is more prevalent in preterm infants. However, there are cases in term infants, but usually, they are associated with predisposing disorders. In this case report, a child is presented with 22q11DS that postnatally developed NEC associated with an adenoviral infection. Although other viruses such as toroviruses or cytomegaloviruses have been implicated in the pathogenesis of NEC in preterm infants, we could not find any report in the recent medical literature describing an association between adenoviral infections, NEC, and 22q11DS in a term infant. PMID- 18405700 TI - Perforated appendix in hernial sac mimicking torsion of undescended testis in a neonate. AB - In pediatric surgical practice, finding of the vermiform appendix in an inguinal hernia sac is not that uncommon; however, a perforation is rare. There are only a few case reports of a perforated appendix with periappendicular abscess in the inguinal hernial sac in a neonate. We report an unusual case of inguinal hernia containing a perforated appendix that was clinically mimicking testicular torsion of the undescended testis. PMID- 18405701 TI - Is extracorporeal shock wave lithotripsy in pediatrics a safe procedure? AB - Removal of urinary calculi is an essential element in the successful treatment of patients with urinary stone disease. The new generation of lithotriptors allows the treatment without the need for general anesthesia. The patients, often outpatients, have a faster discharge from the hospital with a reduction of hospitalization time and operating costs. Shock wave lithotripsy (SWL) is currently considered a safe technique for treatment of pediatric urinary lithiasias, with a low percentage of complications and subsequent surgical retreatments. But can we define SWL as a safe procedure in pediatrics? Herein, we will review the literature to justify SWL safety in children, focusing on important parameters as the insertion of preoperative stenting, side effects, and complications after the procedure. PMID- 18405702 TI - Long gap esophageal atresia: an Australian experience. AB - BACKGROUND: The diagnosis of long gap esophageal atresia (LGEA) may preclude immediate primary anastomosis. We reviewed our experience with this entity for a period of 10 years. METHODS: A retrospective review was undertaken of the medical records of all patients managed for esophageal atresia (EA)/tracheoesophageal fistula (TEF) during the period from 1991 to 2001 at the Children's Hospital at Westmead, Sydney, Australia. Esophageal atresia was defined as long gap when primary repair was considered technically impossible by the surgeon. Also, a questionnaire was sent to all the general pediatric surgeons in Australia to explore their attitude toward LGEA management. RESULTS: One hundred three patients with EA were managed for that period, 17 (16%) of them were defined as LGEA, with mean gap of 5 cm (SD, 1cm). Eight patients (47%) had TEF. Sixteen patients had gastrostomy tube (GT) insertion at a mean age of 4 days. Six patients had esophagostomy at a mean age of 27 days. Thirteen patients had EA repair at a mean age of 146 days. Four patients died before repair and 2 after repair secondary to associated anomalies. Fifty percent of Australian Pediatric Surgeons (APS) responded to the questionnaire. Forty percent defined LGEA as a gap more than 3 to 4 vertebral bodies, whereas 24% considered the absence of TEF as an indication of LGEA. Fifty-six percent of APS will measure the gap on preoperative chest x-rays, and 80% will assess the gap at thoracotomy for ligation of TEF. Ninety-two will measure the gap by inserting a bougie into the upper pouch and into the lower esophagus via the GT. If LGEA was diagnosed, all APS will perform GT with delayed repair. Seventy-two percent of APS will attempt delayed primary repair within 3 to 6 months of age. Seventy-six percent will perform hiatal dissection, and 48% will use upper pouch myotomies. Forty-eight percent will perform gastric pull up, and 32% will use gastric tube for esophageal replacement. CONCLUSIONS: Long gap esophageal atresia represents a surgical challenge. Mortality rate is high secondary to associated anomalies. There is no consensus among APS regarding the definition of LGEA. In general, the consensus of APS would be that the preservation of the patient's own esophagus should be attempted before considering the use of an esophageal replacement. PMID- 18405703 TI - Cancer after repair of esophageal atresia: population-based long-term follow-up. AB - OBJECTIVE: Esophageal atresia (EA) predisposes to gastroesophageal reflux that is associated with intestinal metaplasia and may result in development of adenocarcinoma of the esophagus. To date, the literature has reported 5 cases of esophageal cancer in adult patients treated for EA. The aim of this study was to find out the incidence of esophageal cancer in adult patients with repaired EA. PATIENTS AND METHODS: Five hundred two consecutive patients treated for EA from 1949 to 1978 were followed-up for cancer through the files of the population based countrywide cancer registry from 1967 to 2004. The number of cancer cases observed and person-years at risk were counted, and the expected number of cancer cases estimated from the national cancer incidence rates. The standardized incidence ratios (SIRs) were calculated by dividing the number of cancer cases observed by the expected numbers. RESULTS: None of the 502 patients were lost to follow-up; 230 patients who died before 1967 younger than the median age of 8 days were excluded from further analysis. The 272 remaining patients (142 males) were eligible for follow-up (median age, 35 years; range, 2 days to 56 years). Three cases of cancer were found (SIR, 1.0; 95% confidence interval, 0.20-2.8). One was lymphoma in small intestine, 1 was leukemia, and 1 carcinoma of the uterus but no cases of esophageal cancer. CONCLUSIONS: Our study is able to exclude long-term risk for esophageal cancer after repair of EA 500-fold higher than that of the normal population. Considering the relatively young age of the survivors, further studies and continued follow-up are warranted to elucidate risk for esophageal cancer and need for endoscopic surveillance after repair of EA. PMID- 18405704 TI - Predicting perinatal outcome in isolated congenital diaphragmatic hernia using fetal pulmonary artery diameters. AB - OBJECTIVE: The aim of the study was to evaluate the potential of fetal pulmonary artery (PA) diameters to predict perinatal death and pulmonary arterial hypertension (PAH) in congenital diaphragmatic hernia (CDH). STUDY DESIGN: In this prospective observational study, observed PA (main, right, and left) diameters were measured at the level of the 3 vessels in 21 fetuses with isolated CDH and in 85 controls at 22 to 36 weeks. The observed/expected (o/e) diameters of the main, contralateral, and ipsilateral PAs were calculated by comparing these measurements with reference values obtained in our previous study and correlated with perinatal death and postnatal PAH. RESULTS: The o/e PA diameters were significantly reduced in fetuses with CDH compared to controls (P < .001) and in fetuses with CDH who died (P < .050). However, there was no significant association between PA diameters and PAH (P >or= .050). CONCLUSIONS: The PA diameters might be useful to predict perinatal death in isolated CDH but not postnatal PAH, suggesting that PA diameters are probably related to the severity of pulmonary hypoplasia. PMID- 18405706 TI - Main bronchus occlusion for treatment of congenital diaphragmatic hernia in fetal lambs. AB - PURPOSE: The present study investigates a new surgical approach in the treatment of left diaphragmatic hernia in fetal sheep. We postulated that unlike tracheal occlusion where both lungs are occluded, selective left main bronchus occlusion (LMBO) would accelerate growth of only the left lung, reduce left visceral herniation, and recover space in the both thoraces necessary for lung development. METHODS: Left-sided congenital diaphragmatic hernia (CDH) was surgically created in 8 fetal lambs at approximately 65 days of gestation; in 4 of these animals, LMBO was performed at approximately 118 days. Four sham operated animals served as controls. Lambs were delivered by hysterotomy at 137 days. We measured lung-to-body weight ratios, alveolar surface area, septal wall thickness, and AE2 cell density in the left and right lungs. RESULTS: Left main bronchus occlusion increased (P < .05) left lung growth causing severe right mediastinal shift but failed to reduce herniated abdominal viscera in 3 of 4 lambs. Wet lung-to-body weight ratio in LMBO group was similar to that of the control group; however, this was achieved by overgrowth of left lung, whereas the right wet lung-to-body weight ratio remained low. In the LMBO group, right lung AE2 cell density was higher than that of control group and not different to that of the CDH group. CONCLUSIONS: Left main bronchus occlusion failed to restore normal pulmonary development in CDH. PMID- 18405705 TI - Hirschsprung disease: do risk factors of poor surgical outcome exist? AB - INTRODUCTION: Long-term results after surgical treatment of Hirschsprung disease (HSCR) are not always as good as a surgeon may perceive. Several studies have tried to correlate preoperative features to the surgical outcome of HSCR, but none came to definitive conclusions. Our study is aimed at identifying risk factors of poorer long-term outcome after surgery for HSCR. MATERIALS AND METHODS: One hundred two patients with HSCR were included. Eighty had rectosigmoid aganglionosis and 22 had total colonic aganglionosis (TCSA). Preoperative variables were sex, associated anomalies, delayed meconium passage, preoperative enterocolitis, preoperative bowel obstruction, age at surgery, and number of pull-through procedures performed. Outcome measures were surgical complications, postoperative enterocolitis, perineal excoriations, constipation, continence, psychological self acceptance, and patients' perspectives. These were evaluated with regard to preoperative features and length of aganglionosis. RESULTS: Results of patients with TCSA proved to be significantly worse than those of patients with classic HSCR (constipation and poor continence excluded). Male patients with TCSA proved to have a significantly higher incidence of complications (100% vs 38.5%) and poor psychological self acceptance (100% vs 46.2%). Patients with associated anomalies (central nervous system) experienced a significantly higher incidence of postoperative constipation (43% vs 13%). Patients who described failure or delayed meconium passage complained of significantly worse continence (28.3% vs 7.1% of fair to poor continence). Preoperative enterocolitis proved to be significantly correlated to postoperative enterocolitis (16.4% vs 37.1%) and perineal excoriations (13.7% vs 30.9%). Patients with preoperative intestinal obstructions complained of significantly worse psychological self acceptance (37.8% vs 12.5%). CONCLUSIONS: Length of aganglionosis has the greatest impact on overall surgical outcome of HSCR. Other minor risk factors have been identified, namely, male sex, associated central nervous system anomalies, failure to pass meconium, and preoperative enterocolitis occurrences. Age at surgery and redo procedures proved not to significantly interfere with the outcome. Our study provides comprehensive and useful data to inform parents and families of a baby with HSCR with regard to expected results and long-term outcomes of surgery basing on preoperative features. Further studies on larger series are strongly recommended. PMID- 18405707 TI - Perinatal profile of ventricular overload markers in congenital diaphragmatic hernia. AB - BACKGROUND: In congenital diaphragmatic hernia (CDH), pulmonary hypertension increases right ventricle (RV) afterload, which could impair heart function and contribute to poor outcome for most affected infants. Nevertheless, the real significance of vascular pulmonary alterations in perinatal hemodynamics is largely unknown. It is defined that ventricular pressure overload induces increased myocardium gene expression of B-type natriuretic peptide (BNP) and components of the renin-angiotensinogen and endothelin (ET)-1 systems. Our aim was to evaluate perinatal myocardium expression of these genes associated with ventricular pressure overload in a nitrofen-induced CDH rat model. METHODS: In the nitrofen-induced CDH rat model, fetuses from dated pregnant Sprague-Dawley rats at 15.5, 17.5, 19.5 and 21.5 days postcoitum as well as newborn pups were assigned to 3 experimental groups: control, nitrofen (exposed to nitrofen, without CDH), and CDH (exposed to nitrofen, with CDH). Myocardial samples collected from the RV and left ventricle (LV) were processed for quantification of messenger RNA (mRNA) of BNP, angiotensinogen, and ET-1. RESULTS: The perinatal expression of BNP, angiotensinogen, and ET-1 mRNA in the RV and LV of the control group revealed daily changes. During gestation, the expression of BNP and angiotensinogen mRNA underwent significant oscillation compared with control in both nitrofen-exposed fetuses, although we cannot identify significant differences between the nitrofen and CDH groups. After birth, we found a significant increasing expression of all studied genes only in the RV of CDH pups. CONCLUSIONS: Perinatal myocardial quantification of BNP, angiotensinogen, and ET-1 mRNA levels suggests that both nitrofen-exposed and control pups revealed prenatal variations of expression of the studied genes. Moreover, CDH is associated with significant molecular alterations only in the RV after birth. PMID- 18405708 TI - Tailored resective pancreatic surgery for pediatric patients with chronic pancreatitis. AB - BACKGROUND: Surgical treatment for chronic pancreatitis (CP) in children comprises predominantly nonresective draining procedures. The purpose of this study was to identify indications, techniques, and results of organ-preserving resective pancreatic procedures for pediatric CP at our institution. PATIENTS AND METHODS: A retrospective chart review was performed of all children undergoing pancreatic surgery for CP over a period of 4 years. RESULTS: Overall, 6 pediatric patients (3 male, 3 female, ages 7-18 years) underwent a duodenum-preserving pancreatic head resection (3), a middle segmental pancreatic resection (2), or a distal pancreatectomy (1) for CP of different etiologies (idiopathic 2, posttraumatic 2, pancreas divisum 1, situs inversus 1). No mortality or major surgical complication occurred. Mean operative time was 294 min (207-412 min) and intraoperative blood loss was 541 mL (100-1300 mL). Postoperative hospital stay was 13 days (10-18 days). No endocrine or exocrine insufficiency occurred during follow up of 46 months (25-50 m), and pain control was improved in 5 of 6 patients. CONCLUSIONS: Tailored organ-preserving resective pancreatic surgery can be performed with low morbidity and mortality in pediatric patients with CP and not responding to conservative treatment. PMID- 18405709 TI - Laparoscopic correction of biliary duct stenosis in choledochal cyst. AB - BACKGROUND/PURPOSES: Laparoscopy has been widely accepted as a technique for the excision of choledochal cyst, but there has been little experience using it as a therapeutic modality for hepatic duct stenosis. The aim of this study is to present our experiences in laparoscopic excision of biliary stenosis and Roux-en Y reconstruction for patients with choledochal cysts. METHODS: Eight patients, 3 boys and 5 girls (ranged from 6 months to 12 years; median age, 3.6 years), with hepatic duct stenosis underwent laparoscopic excision of the cyst and ductoplasty, with a Roux-en-Y hepaticoenterostomy between July 2001 and January 2005. Seven of the 8 patients had common hepatic duct stenosis with intrahepatic duct dilatation, and 1 had right hepatic duct stenosis with proximal dilatation. Four ports were inserted for instruments of 3- and 5-mm sizes. Each patient underwent a laparoscopic cholangiography. The gallbladder and dilated bile ducts were completely excised. The strictures of the hepatic ducts were treated by ductoplasty. The cut end of the duct was widened by incising along the anterior wall of the hepatic duct after excision of the narrow segment. By using intraoperative bile duct endoscopy, the optimal level of resection of the common hepatic duct was determined safely without endangering the orifices of the hepatic ducts or leaving any redundant duct, and the stone debris in the bile duct was identified and washed out. The Roux-en-Y jejunal loop was fashioned extracorporeally by exteriorizing the jejunum through the umbilical incision (1.0 1.5 cm) and passed up retrocolically followed by an end-to-side hepaticojejunostomy. RESULTS: The median duration of the operation was 4.3 hours (3.8-5.6 hours). Intraoperative bleeding was minimal, with no patients requiring blood transfusion. In 7 of the 8 cases, postoperative hospital stay ranged from 4 to 6 days; the other case had bile leak, which was cured by draining for 26 days without surgical intervention. All the patients had been followed up for 6 to 51 months. They stayed asymptomatic and well with no delayed complication. CONCLUSIONS: Laparoscopically assisted hepatic ductoplasty is effective and safe for children with choledochal cyst. Bile duct endoscopy proved to be a valuable instrument in showing detailed variations of the biliary system and allowed a safe hepatic hilum exploration and accurate placed hepaticojejunal anastomosis. PMID- 18405710 TI - Prenatal ultrasonographic gastrointestinal abnormalities in fetuses with gastroschisis do not correlate with postnatal outcomes. AB - PURPOSE: In the setting of gastroschisis, the clinical significance of prenatal ultrasound findings of secondary changes in bowel appearance remains unknown. The purpose of this study was to correlate prenatal identification of additional gastrointestinal sonographic abnormalities with postnatal clinical outcome. METHODS: A retrospective review was conducted on 64 fetuses with a prenatal diagnosis of gastroschisis treated at the Children's Hospital of Philadelphia from 2000 to 2007. Postnatal outcomes were compared between newborns with additional sonographic gastrointestinal abnormalities and those without secondary changes to the bowel appearance. RESULTS: Thirty (47%) patients had at least one gastrointestinal abnormality (eg, bowel dilatation, echogenic bowel, thickened bowel, matted bowel, herniation of the stomach through the abdominal wall defect, or segmental loss of bowel peristalsis) on prenatal ultrasound. There were no significant differences between groups with respect to the time to initial and full enteral nutrition, total hospital stay, requirement for ventilator support, central line infection rates, reoperation rates, or mortality. CONCLUSIONS: In the setting of gastroschisis, isolated findings of gastrointestinal abnormalities on prenatal ultrasound do not correlate with adverse postnatal outcome. PMID- 18405711 TI - Gastroschisis: the cost of an epidemic. AB - BACKGROUND: The rising incidence of gastroschisis has been highlighted by the Department of Health as a growing concern. As well as the health implications for the increasing number of affected infants, this increase in incidence will have an impact of the costs of health care. This study was undertaken to estimate the financial cost of treating this condition in one tertiary neonatal surgical center. METHODS: A retrospective analysis was performed of all patients admitted to a tertiary neonatal surgical center with gastroschisis from January 1996 to December 2005. The main outcome measures were incidence, length of hospital stay, and total cost for all patients each year. RESULTS: The incidence of gastroschisis has risen 3-fold in 10 years. The median cost per patient is relatively constant. A few patients with severe intestinal dysmotility require prolonged hospital stay. As the condition becomes more common, there are an increasing number of complex patients and thus an increase in annual costs, which is disproportionate to the increase in numbers of cases. We estimate that the annual cost to the National Health Service (NHS) of this condition in England and Wales has risen from pound3.6 million in 1996 to in excess of pound15 million in 2005. CONCLUSIONS: Urgent research is required into the etiology of gastroschisis and into the severe intestinal dysmotility that occurs in some complex patients. PMID- 18405712 TI - The functional sequelae of sacrococcygeal teratoma: a longitudinal and cross sectional follow-up study. AB - AIM: The purpose of the study was to determine clinical manifestations, prevalence, and natural history of functional sequelae in patients operated on for sacrococcygeal teratoma (SCT) during infancy. METHODS: The medical records of 18 infants operated on for SCT were reviewed. Data recorded during admission and during routine outpatient appointments were analyzed. Moreover, 13 surviving patients (mean age, 25 years) and 65 age- and sex-matched controls were evaluated at a special outpatient clinic. RESULTS: Seven adult patients and 25 controls presented with one or more functional disorders including abdominal pain, constipation, diarrhea, soiling, enuresis, urinary frequency, urinary stress incontinence, and urinary straining. In 7 of 8 symptomatic children, one or more functional disorders disappeared during adult life. In comparison with controls, adult patients had a nonsignificant higher prevalence of each individual dysfunction. CONCLUSIONS: Sequelae of SCT tend to improve with time, and the disorders reported are relatively common complaints in the general population. PMID- 18405713 TI - Neonatal repair of cleft lip: a decision-making protocol. AB - PURPOSE: Treatment of clefts lip during the neonatal period remains a controversial subject. Those who are in favor of delayed closure argue a higher risk general anesthesia when it was performed in neonatal period. The purpose of this study was to evaluate the complications and the feasibility of this surgery during the neonatal period. METHODS: This was a retrospective study of 61 children with labial, labioalveolar, labio-alveolo-palatine, and labiopalatine clefts between May 2000 and November 2006. Each patient's medical file and particularly his or her anesthesia file was used to record the principal demographic data, the results of the malformation workup, and preoperative complications. RESULTS: Sixty-one newborns, 20 girls and 41 boys, aged 7.5 +/- 6.7 days were operated on. The mean weight on the day of surgery was 3190 +/- 454 g. Fifty-four children had a malformation workup (abdominal ultrasonography, spinal bone workup, transfontanelle ultrasonography, and cardiac ultrasonography). Thirteen associated malformations (21%) were thereby detected. There were no surgical complications. The anesthesiologists did not have any real intubation problems. In 4 cases, however, intubation was only possible after several laryngoscopies and changing the type of intubation shaft. There were no major complications. However, one child did present a preoperative complication. It was an episode of desaturation with bradycardia that was quickly resolved without further consequences in a child with a ventricular septal defect and an auricular septal defect. CONCLUSIONS: We think that neonatal lip closure should continue to be performed. It is essential for the psychological status of the parents. We have not found any studies in the literature that reported an anesthesia risk that was greater in the neonatal period than at 3 months in patients without risk of complications. PMID- 18405714 TI - Speech prognosis and need of pharyngeal flap for non syndromic vs syndromic Pierre Robin Sequence. AB - BACKGROUND: The aim of this retrospective study was to evaluate speech outcome and need of a pharyngeal flap in children born with nonsyndromic Pierre Robin Sequence (nsPRS) vs syndromic Pierre Robin Sequence (sPRS). METHODS: Pierre Robin Sequence was diagnosed when the triad microretrognathia, glossoptosis, and cleft palate were present. Children were classified at birth in 3 categories depending on respiratory and feeding problems. The Borel-Maisonny classification was used to score the velopharyngeal insufficiency. RESULTS: The study was based on 38 children followed from 1985 to 2006. For the 25 nsPRS, 9 (36%) pharyngeal flaps were performed with improvements of the phonatory score in the 3 categories. For the 13 sPRS, 3 (23%) pharyngeal flaps were performed with an improvement of the phonatory scores in the 3 children. There was no statistical difference between the nsPRS and sPRS groups (P = .3) even if we compared the children in the 3 categories (P = .2). CONCLUSIONS: Children born with nsPRS did not have a better prognosis of speech outcome than children born with sPRS. Respiratory and feeding problems at birth did not seem to be correlated with speech outcome. This is important when informing parents on the prognosis of long-term therapy. PMID- 18405716 TI - The dysraphic levels of skin and vertebrae are different in mouse fetuses and neonates with myelomeningocele. AB - BACKGROUND: Mouse fetuses with spontaneous myelomeningocele (MMC) were investigated, determining the various levels of dysraphism in soft tissue, spinal cord, and vertebrae. Morphology was correlated with hind limb function. METHODS: Viable curly tail/loop tail mouse fetuses underwent qualitative standardized ex utero examination of tail and hind limb sensitivity and motor response. Afterward, they were processed either for histology or skeletal preparation. RESULTS: All animals displayed identical cranial levels of soft tissue and neural defects. The cranial opening of the vertebral defects were invariably located more cranially (range, 0.5-5 vertebrae; mean = 2.25). The caudal opening of soft/neural tissue and bony defects was invariably at the coccygeal base. The comparison of functional with morphological levels demonstrated that, in 52.5%, the level of the soft/neural tissue dysraphism and, in 47.5%, the level of the bony opening correlated with the neurologic deficit. CONCLUSION: The naturally occurring soft tissue coverage over the MMC could exert a protective effect toward the underlying spinal cord. This interpretation supports the concept that in utero acquired destruction of exposed neural tissue is a main factor for the neonatal functional deficit. Thus, these data are consistent with the rationale for prenatal MMC repair in humans. PMID- 18405715 TI - Fetal wound healing using a genetically modified murine model: the contribution of P-selectin. AB - PURPOSE: During early gestation, fetal wounds heal with paucity of inflammation and absent scar formation. P-selectin is an adhesion molecule that is important for leukocyte recruitment to injury sites. We used a murine fetal wound healing model to study the specific contribution of P-selectin to scarless wound repair. METHODS: Linear excisional wounds were created on the dorsa of E15.5 and E17.5 gestation fetuses in wild-type and P-selectin (-/-) mice (term = 19 days). Wounds were harvested at various time-points after wounding and analyzed using histology and immunohistochemistry. RESULTS: The E15.5 wounds in both wild-type and P selectin (-/-) fetuses healed scarlessly and with minimal inflammation, whereas E17.5 wounds healed with fibrosis and inflammation. However, the scars of the P selectin (-/-) wounds appeared slightly different than wild-type. There were significantly more inflammatory cells in E17.5 wild-type wounds 6 hours after injury (P < .001), but the difference was no longer significant by 24 hours. Finally, reepithelialization was slower in the E15.5 knockout wounds compared to their wild-type counterparts. CONCLUSIONS: Absence of P-selectin delays inflammatory cell recruitment and reepithelialization of fetal wounds; however, scar formation still occurs in late gestation wounds. The contribution of specific molecules to fetal wound healing can be elucidated using murine knockout or transgenic models. PMID- 18405717 TI - The demographics of playground equipment injuries in children. AB - BACKGROUND/PURPOSE: There have been many different studies of injuries owing to playground equipment but none that have looked in detail using large nationwide databases. It was the purpose of this study to investigate injuries owing to playground equipment using the National Electronic Injury Surveillance System (NEISS) database and further understand their demographics. METHODS: Detailed NEISS injury data from 2002 through 2004 for slides, monkey bars, and swings were analyzed. Appropriate statistical analyses were performed; because of the many analyses on this large data set, P < .01 was considered statistically significant. RESULTS: There were 22728 emergency department visits owing to playground equipment injuries recorded by NEISS between 2002 and 2004; 83.9% were owing to monkey bars, swings, and slides, and the 5 most common diagnoses were fractures (39.3%), contusions/abrasions (20.6%), lacerations (16.6%), strains/sprains (9.9%), and traumatic brain injuries (TBI) (8.5%). There were 9487 boys (54.3%) and 7995 girls (45.7%). The average age was 6.5 +/- 3.0 years. The injuries occurred at school in 38.9%; at a recreation/sporting facility, in 35.5%; and at home, in 25.6%. Most were treated and released (94.4%). Amerindian children were 2 times more likely than blacks to be admitted; compared to contusions, fractures were 9.8 times, and TBIs, 4.7 times more likely to be admitted. Injuries on monkey bars were 1.2 times more likely to be admitted than those on swings or slides. Fractures were 1.9 times more likely to occur on a monkey bar compared with swings or slides. Traumatic brain injuries were 1.4 times more likely to occur on a swing compared to slides or monkey bars. CONCLUSION: Swings at school are the most common mechanism of injury for TBIs, and the seasonal data would suggest that increased supervision of children using swings during school hours might reduce the occurrence of TBIs. Monkey bars are the most common cause of fracture, and fracture is the most common cause of admission. Prevention strategies to reduce the number of fractures should be directed at monkey bar equipment and landing surfaces. The trend in playground equipment injury also indicates that monkey bars are problematic because the number of injuries per year per capita owing to monkey bars is stable, whereas those from swings and slides is decreasing. PMID- 18405718 TI - Bone health in patients with cloacal exstrophy and persistent cloaca after bladder augmentation. AB - PURPOSE: In this study, the prevalence of osteoporosis was evaluated in patients operated on for cloacal exstrophy or persistent cloaca and who had undergone bladder augmentation. METHODS: Seven patients operated on because of cloacal exstrophy and 3 patients operated on because of persistent cloaca were evaluated. Patients were clinically assessed. Areal bone mineral density for the lumbar spine (L1-L4), femoral neck, and whole body was measured with dual-energy x-ray absorptiometry. Acid-base balance, plasma 25-hydroxyvitamin D (vitamin D), and gonadal function were determined and correlated with dual-energy x-ray absorptiometry results. RESULTS: The patients were small in height (median, -2.2 SD). According to areal bone mineral density measurements, 40% of the patients had significantly reduced BMD. When the values were adjusted for patient size, only 1 patient had slightly reduced bone mass. Half of the patients had low vitamin D values. The height-corrected bone mineral content values showed a positive correlation with vitamin D values (P = .0072) but were not different in patients with and without hypogonadism (P = .33). CONCLUSIONS: Patients with cloacal exstrophy or complicated cloaca have many risk factors for osteoporosis. However, this study showed good overall bone health in affected adolescents. PMID- 18405719 TI - Ethyl pyruvate reduces germ cell-specific apoptosis and oxidative stress in rat model of testicular torsion/detorsion. AB - PURPOSE: Testicular torsion/detorsion (T/D) results in enhanced formation of free radical metabolites, which contributes to the pathophysiology of tissue damage. We investigated the protective effects of ethyl pyruvate (EP) against testis tissue damage in an experimental model of testicular torsion. METHODS: Sprague Dawley rats were divided into 5 groups. In those animals that underwent T/D, right testes were rotated 720 degrees for 1 hour. Group 1 control rats underwent sham operation. In group 2, the rats underwent T/D. The EP was prepared and injected in the form of Ringer's ethyl pyruvate solution. The rats in group 3, 4, and 5 received 2 doses of 20, 50, and 100 mg/kg EP (30 minutes before and after detorsion), respectively. The right testes of 6 animals from each group were excised 4 hours after detorsion for the measurement of lipid peroxidation, myeloperoxidase (MPO), and antioxidant enzymes activities. Germ cell apoptosis was determined in right testes of 8 animals per group 24 hours after detorsion. The epididymal sperm concentration and motility were evaluated 1 month after treatments. RESULTS: Germ cell apoptosis indices were significantly higher in group 2 compared with control group. The level of lipid peroxidation and MPO activity increased, whereas antioxidant enzymes activities decreased after T/D. Sperm count and motility were also reduced 1 month after T/D in group 2 rats. However, EP treatment at doses of 50 and 100 mg/kg significantly decreased the early apoptotic damage and improved long-term sperm count and motility. In the same dosing groups, we observed normalization of oxidant/antioxidant balance and decrement of MPO activity. However, administration of 20 mg/kg of EP conferred no protective effect. CONCLUSIONS: Administration of Ringer's ethyl pyruvate solution (in appropriate doses) is protective against apoptotic tissue damage following testicular torsion and improves long-term testicular function. The antioxidant and anti-inflammatory properties of EP seem responsible for the protective effects. Our findings suggest this resuscitation solution as a possible substitute for fluid and electrolyte maintenance during surgical detorsion. PMID- 18405720 TI - Alterations in the mechanical properties of bladder smooth muscle in hydrocephalus rat model. AB - OBJECTIVES: It is now well established that hydrocephalus is associated with impaired bladder function. The aim of this study was to determine the effects of hydrocephalus on bladder smooth muscle (BSM) reactivity in the rat model. MATERIALS AND METHODS: Hydrocephalus was induced in 7-day-old rats by injection of kaolin into the cisterna magna (AH group). Control group rats underwent a sham operation. After 10 days, rats were decapitated. Each bladder was excised and BSM strips placed in an organ bath where contractile and relaxant responses were studied. RESULTS: Contractile response of BSM to KCl decreased in the AH group. Increased response to muscarinic agonist carbachol was observed in the AH group. The relaxant response to adrenergic agonist isoprenaline was significantly decreased in the AH group, whereas non-receptor-dependent agonist papaverine was unchanged in 2 groups. CONCLUSION: Bladder smooth muscle reactivity is affected by the formation of hydrocephalus essentially by both receptor-dependent and non receptor-dependent mechanisms. This pathway may be a novel target for the pharmacologic treatment of bladder dysfunction secondary to hydrocephalus. PMID- 18405721 TI - Therapeutic experience from 1428 patients with pediatric tracheobronchial foreign body. AB - PURPOSE: Tracheobronchial foreign body (TFB) aspiration is a life-threatening emergency for children. Knowing how to reduce the incidence of complications and mortality during the management of TFB is critically important. METHODS AND PATIENTS: Pediatric patients with TFB, who were treated in the Department of Otorhinolaryngology-Head and Neck Surgery, The Affiliated Hospital of Medical College Qingdao University, Qingdao, Shandong Province, China, were included in this analysis during 1985 to 2007. One thousand four hundred twenty-eight patient records with TFB were retrospectively reviewed. This study mainly reported location and type of foreign body, complications, anesthesia methods, and outcome. RESULTS: Among the 1428 patients, 1424 (99.72%) underwent successful removal of a foreign body by bronchoscopy. First-attempt removal was successful in 1347 whose bronchoscopy was done under general anesthesia combined with topical anesthesia and in 65 cases with inhaled anesthesia by ether combined with topical anesthesia, respectively. Second-attempt removal was successful in 12 patients whose bronchoscopy was done under inhaled anesthesia by ether combined with topical anesthesia. Foreign bodies were located in the trachea in 75 cases (5.25%), right bronchial tree in 780 patients (54.62%), left bronchial tree in 567 cases (39.71%), and bilateral bronchial tree in 6 cases (0.42%). Types of foreign body included peanuts (1244 cases, 87.12%), beans (93 cases, 6.51%), and others (91 cases, 6.37%). Tracheotomy was performed in 4 patients. Three patients (0.21%) died in this study. Four patients developed toxicity from tetracaine during topical anesthesia. CONCLUSIONS: Bronchoscopy under general anesthesia augmented with topical anesthesia is a very safe and effective procedure for patient with TFB. Surgeons and anesthetists must be aware of the risk of tetracaine toxicity and other complications. PMID- 18405722 TI - Protective effect of erythropoietin pretreatment in testicular ischemia reperfusion injury in rats. AB - BACKGROUND/PURPOSE: This study was designed to investigate the effects of recombinant erythropoietin (EPO), a hormone widely used for treatment of uremic anemia, in rats subjected to testicular ischemia and reperfusion (I/R). METHODS: Thirty-five male rats were divided into the following: control, sham operated, ischemia (I), I/R, and I/R + EPO groups. In the I group, 2 hours of left unilateral testicular torsion were performed, and in the I/R and I/R + EPO groups, an additional 2 hours of testicular detorsions were performed. The I/R + EPO group was pretreated intraperitoneally with EPO (500 IU/kg) before reperfusion. Testicular tissue samples were examined for biochemical and histopathologic parameters. Apoptotic cells in all testes were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling technique and caspase 3 immunohistochemistry. RESULTS: At histopathologic examination, ischemic changes in primary spermatocytes were noted in all torted testes. Cellular damage and apoptosis were more severe in ischemic groups than the EPO pretreated group. There were statistically significant differences in tissue biochemical parameters in the I and I/R groups compared with the I/R + EPO group. CONCLUSIONS: The results of the present study suggest that EPO exerts protective effects against I/R injury via the modulation of free radical scavenger's activities, which decreases lipid peroxidation levels and attenuation of apoptosis. PMID- 18405723 TI - Giovanni Battista Morgagni and his contribution to pediatric surgery. AB - Pediatric surgery is a less than a century old surgical specialty. However, early knowledge of human malformations and pediatric surgical conditions dates back to centuries before. One of the main contributors to progress in these fields was Giovanni Battista Morgagni (1682-1771), who also had a substantial weight in the development of future pediatric surgery. With his masterpiece De Sedibus et Causis Morborum per Anatomen Indagatis (On the Seats and Causes of Diseases Investigated by Anatomy) (1761), he set the basis for modern pathology. In this textbook, Morgagni was the first to describe anatomical elements like the trigonum sternocostale dextrum (the Morgagni's foramen), the appendix testis (the Morgagni's hydatid), and the vertical folds of distal rectum (the Morgagni's columns). He was also the first to describe pediatric pathological conditions like epispadia, meconium peritonitis, Crohn's disease, and coarctation of the aorta. Finally, he substantially contributed to the understanding of the pathophysiology of conditions like the vesico-ureteral reflux, the anterior wall defects (gastroschisis/omphalocele), and the spina bifida. For this, it was said of him: "If all the anatomical findings made by Morgagni should bear his name, probably one third of human body would be called Morgagni's." PMID- 18405724 TI - The best 10 clinical articles for the last 50 years from the Division of General Surgery, Hospital for Sick Children, Toronto, Ontario, Canada. AB - PURPOSE: The aim of the review was to present the best 10 clinical articles for the last 50 years (1956-2006) from the Division of General Surgery, Hospital for Sick Children (HSC), Toronto, Canada. These articles were judged by the major change (impact) in clinical practice of pediatric general surgery after their publication. METHODS: All clinical articles from 1956 to 2006 inclusively written by members of the division (while working at HSC) were evaluated. The 2 authors of this article (retired honorary staff surgeon and recently trained chief surgical resident/fellow) separately rated the articles. Each lead author (if possible) was asked to comment on "the significance of their paper, then and now." If the lead author was unavailable, 1 of the 2 authors of this article commented on the articles. RESULTS: The best 10 clinical articles selected involved spleen trauma, necrotizing enterocolitis, esophageal replacement, Hirschsprung's disease, tracheal compression, fecal incontinence, gastroesophageal reflux, diaphragmatic hernia, and ruptured appendix. There were 8 staff members and 5 surgical residents/fellows who were lead authors, along with 10 staff from other divisions, departments, and/or hospitals. CONCLUSION: The conservative management of splenic trauma was judged the best article from this Division that made the largest clinical impact for the last 50 years. PMID- 18405726 TI - Association of Shah-Waardenburgh syndrome: a review of 6 cases. AB - Shah-Waardenburg syndrome (SWS) is a neurocristopathy and is characterized by Hirschsprung's disease (HD), deafness, and depigmentation of hairs, skin, and iris. OBJECTIVE: The aim of the article is to study the relative frequency of associations in 6 consecutive cases of SWS. METHODS: A review of 6 consecutive patients with SWS was performed to study the frequency of various components of the syndrome. RESULTS: Six patients had features of SWS. All patients had HD; of these, 3 had rectosigmoid HD, whereas 3 had extended HD. All patients had white forelock of hairs with skin depigmentation. One patient had sensorineural deafness, whereas other babies were less than 1 year, and thus, full evaluation of hearing deficiency was not assessed. Three patients had blue eyes, whereas other babies had normal iris pigmentation. Skin depigmentation was noted in 5 of the 6 patients. Three babies were seriously malnourished and showed higher association of enterocolitis. CONCLUSION: Shah-Waardenburg syndrome is an uncommon association of HD. Depigmentation with a white forelock and skin manifestations are common, whereas blue iris, long segment disease, and enterocolitis are present in nearly half of the patients. PMID- 18405725 TI - Tension gastrothorax: a case report and review of literature. AB - BACKGROUND: Tension gastrothorax develops when the stomach, herniated through a congenital diaphragmatic defect into the thorax, is massively distended by trapped air. We report a case of tension gastrothorax and review the literature. CASE REPORT: A previously healthy 8-month-old female presented with severe respiratory distress, misdiagnosed as tension pneumothorax. Intercostal tube was inserted. The tube was noted to drain food as well as air. The patient was investigated by radio-contrast swallow, which demonstrated the presence of the stomach in the chest. The patient was operated upon, and the stomach, transverse colon, and spleen were reduced back to the abdomen. The defects in the stomach and diaphragm were closed. CONCLUSION: Tension gastrothorax is a life-threatening condition leading to acute and severe respiratory distress. The presence of air filled structure in left hemithorax in a previously healthy child presenting with acute respiratory distress should prompt the inclusion of tension gastrothorax in the differential diagnosis. PMID- 18405727 TI - Double cervical cyst derived from a single thyroglossal duct tract. AB - Thyroglossal duct cysts are one of the most common congenital cervical pathologic findings in children. This type of cyst can be located anywhere between the base of the tongue and the sternal manubrium. We report the case of a patient with a double thyroglossal cyst, one located in the hyoid region and the other in the thyroid gland. The 2 cysts were connected by a permeable tract, which confirms that an involution failure of the embryonic remains of the thyroglossal duct was responsible for the development of the cyst. Intrathyroid cysts of the thyroglossal duct, though uncommon, should be included in the differential diagnosis of thyroid cysts in children. PMID- 18405728 TI - Giant hepatic adenoma with atypical features in a patient on oxcarbazepine therapy. AB - An association between oxcarbazepine therapy and hepatic adenoma (HA) has been documented in animal models but not observed in humans. The authors report a case of a 16-year-old girl on oxcarbazepine therapy for seizure disorder who presented with a giant HA. Pathology of the HA was notable for marked periductal fibrosis and glycoprotein inclusions in the nontumor liver. The patient was not on oral contraceptives and has no other known risk factors for HA. PMID- 18405729 TI - Newborn and toddler intestinal obstruction owing to congenital mesenteric defects. AB - Transmesenteric hernia is a rare cause of intestinal obstruction most commonly affecting the small bowel. The mesenteric defect is usually 2 to 3 cm in diameter. The authors describe 2 cases of young pediatric patients presenting with bowel obstruction resulting from a congenital mesenteric defect. The initial patient had a 30-cm-wide congenital defect in the ileal mesentery through which the sigmoid colon and some loops of small bowel had herniated. The second patient is a newborn infant who presented with symptoms and radiographic evidence of proximal bowel obstruction initially thought to be resulting from malrotation with midgut volvulus but was found at surgical exploration to have a small defect in the ileal mesentery. PMID- 18405730 TI - Primary parotid non-Hodgkin lymphoma in an infant. PMID- 18405731 TI - Ruptured abdominal aortic aneurysm in an 11-year-old boy. AB - We present a case of a ruptured aortic aneurysm in an 11-year-old boy presenting with loss of consciousness. The presentation, management, pathology, and gravity of this condition are discussed. PMID- 18405732 TI - Distal vaginal agenesis in monozygotic twins: case report. AB - Distal vaginal agenesis is one of the rarest congenital malformations of female genital tract. We describe successfully operated teenaged monozygotic twins by combined abdominal and perineal approach. The imaging studies and surgical experience were presented. PMID- 18405733 TI - Cystic dysplasia of the rete testis. AB - Cystic dysplasia of the rete testis is a rare benign testicular tumor that is found mainly in the pediatric population and that must be distinguished from malignant lesions. Ultrasonography can aid in diagnosis and in evaluation for associated renal lesions. Treatment options include orchiectomy, testicular sparing excision of the cystic mass, and nonoperative surveillance. To increase awareness of the disorder, we present the case of a 2-month-old male infant with cystic dysplasia of the rete testis. PMID- 18405734 TI - Refinements in pectus carinatum correction: the pectoralis muscle split technique. AB - BACKGROUND: The standard approach for correction of pectus carinatum deformity includes elevation of the pectoralis major and rectus abdominis muscle from the sternum and adjacent ribs. A postoperative restriction of shoulder activity for several weeks is necessary to allow stable healing of the elevated muscles. To reduce postoperative immobilization, we present a modified approach to the parasternal ribs using a pectoralis muscle split technique. METHODS: At each level of rib cartilage resection, the pectoralis muscle is split along the direction of its fibers instead of elevating the entire muscle as performed with the standard technique. From July 2000 to May 2007, we successfully used this technique in 33 patients with pectus carinatum deformity. RESULTS: After the muscle split approach, patients returned to full unrestricted shoulder activity as early as 3 weeks postoperatively, compared to 6 weeks in patients treated with muscle flap elevation. Postoperative pain was reduced and the patients were discharged earlier from the hospital than following the conventional approach. CONCLUSIONS: The muscle split technique is a modified surgical approach to the parasternal ribs in patients with pectus carinatum deformity. It helps to maintain pectoralis muscle vascularization and function and can reduce postoperative pain, hospitalization, and rehabilitation period. PMID- 18405735 TI - Single-stage repair for rectovestibular fistula without opening the fourchette. AB - BACKGROUND: Anorectal malformations are one of the most common congenital defects. This study is conducted to demonstrate new technique for treatment of rectovaginal fistula without disturbing the fourchette through posterior sagittal approach. METHOD: All the patients of rectovestibular fistula admitted after the neonatal age were treated with posterior sagittal anorectoplasty without opening the fourchette. The results were evaluated for cosmetic appearance and anal continence. RESULT: A total of 40 patients were included in our study. All patients were more than 1 month old. Operative time ranges from 70 to 150 minutes. The cosmetic appearance was good. Anal continence was good in 72% cases and fair in 20% cases. Fifteen percent of patients had minimal constipation and 7.5% patients had mucosal prolapse. CONCLUSION: Single-stage repair for vestibular anus through posterior sagittal anorectoplasty without opening fourchette has a good cosmetic appearance and good anal continence. PMID- 18405738 TI - Commentary on adjunctive use of testosterone gel with sildenafil for erectile dysfunction. PMID- 18405739 TI - Commentary on the etiologic relationship between prostate radiation and bladder cancer: the scourge for one is poison for the other. PMID- 18405740 TI - Watchful waiting and health related quality of life for patients with localized prostate cancer: data from CaPSURE. AB - PURPOSE: Watchful waiting is an alternative to active treatment for men with low risk prostate cancer but it is unclear how health related quality of life (HRQoL) may change over time for men who select this option. We report on HRQoL in men with localized prostate cancer who selected watchful waiting. MATERIALS AND METHODS: HRQoL outcomes were reviewed for 310 men diagnosed with prostate cancer from 1990 to 2001 within Cancer of the Prostate Strategic Urological Research Endeavor who chose watchful waiting. The UCLA Prostate Cancer Index and RAND 36 Item Health Survey were completed at enrollment and approximately every 6 months. A random slopes model was developed to assess time trends in HRQoL for up to 5 years after diagnosis, adjusting for age at diagnosis and specific comorbidities. RESULTS: Significant decreases with time were observed in 7 domains of the RAND 36-Item Health Survey and 4 of the UCLA Prostate Cancer Index scales. CONCLUSIONS: Men with prostate cancer who chose watchful waiting in the current study had better or similar HRQoL outcomes compared to men without prostate cancer at the start of the study. Many of these scores were significantly affected by increasing age and decreased with time. The physical domain scores as well as sexual function scores decreased more than expected from the aging process alone. PMID- 18405741 TI - Commentary on watchful waiting and health related quality of life for patients with localized prostate cancer. PMID- 18405742 TI - Higher than expected association of clinical prostate and bladder cancers. AB - PURPOSE: In this study we evaluated the risk of a second malignancy of the bladder or prostate in patients with a previous diagnosis of prostate cancer (PCa) or urothelial cancer (TCC). MATERIALS AND METHODS: We retrospectively analyzed all cases of PCa and TCC diagnosed between January 1996 and June 2003. Only PCa diagnosed due to abnormal digital rectal examination or increased prostate specific antigen were included. All patients with TCC presented with hematuria or irritative voiding symptoms and the diagnoses were confirmed with a tissue diagnosis. The incidence of lung, colon and renal cancers was also analyzed. RESULTS: A total of 816 men were diagnosed with PCa and/or TCC. Of 673 men initially diagnosed with PCa 21 had TCC. Of 149 men initially diagnosed with TCC 18 had PCa. Average age at PCa and TCC diagnosis +/- SD was 68.2 +/- 7.9 and 68.2 +/- 10.4 years, respectively. The standardized incidence ratio (SIR) of TCC in patients with PCa (SIR 4.31, 95% CI 2.411 to 7.110) and of PCa in patients with TCC (SIR 3.83, 95% CI 1.911 to 6.858) was significantly increased. There was no statistical significant difference in SIR for TCC in men with or without radiotherapy. SIR for lung, renal or colon cancer was not significantly different from what was expected. CONCLUSIONS: Patients with PCa have higher incidence of bladder cancer and those with bladder cancer have a higher incidence of PCa. This study has clinical implications in the care of these patients and it may stimulate research interest that may identify common pathways of carcinogenesis. PMID- 18405743 TI - 12-year outcomes following permanent prostate brachytherapy in patients with clinically localized prostate cancer. AB - PURPOSE: We reviewed the outcomes in men treated with permanent prostate brachytherapy (PPB). MATERIAL AND METHODS: A total of 1,449 consecutive patients with a mean age of 68 years treated with PPB between 1992 and 2000 and mean pretreatment prostate specific antigen (PSA) 10.1 ng/ml were included in this study. Of the patients 55% presented with Gleason 6 tumors and 28% had Gleason 7 disease. A total of 400 patients (27%) were treated with neoadjuvant hormones and 301 (20%) were treated in combination with external radiation plus PPB. Several biochemical freedom from recurrence (BFR) definitions were determined. Statistical analysis consisted of log rank testing, Kaplan-Meier estimates and Cox regression analysis. RESULTS: Median followup was 82 months with 39 patients at risk at for 144 months. Overall and disease specific survival at 12 years was 81% and 93%, respectively. The 12-year BFR was 81%, 78%, 74% and 77% according to the American Society for Therapeutic Radiology and Oncology (ASTRO), ASTRO Kattan, ASTRO-Last Call and Houston definitions, respectively. The 12-year ASTRO Kattan BFR using risk stratification was 89%, 78% and 63% in patients at low, intermediate and high risk, respectively (p = 0.0001). Multivariate analysis identified the dose that 90% of the target volume received (p <0.0001), pretreatment PSA (p = 0.001), Gleason score (p = 0.002), the percent positive core biopsies (p = 0.037), clinical stage (p = 0.689), the addition of hormones (p = 0.655) and the addition of external radiation (p = 0.724) for predicting BFR ASTRO. Five-year disease specific survival was 44% in patients with a PSA doubling time of less than 12 months vs 88% in those with a PSA doubling time of 12 months or greater (p = 0.0001). CONCLUSIONS: PPB offers acceptable 12-year BFR in patients who present with clinically localized prostate cancer. Implant dosimetry continues as an important predictor for BFR, while the addition of adjuvant therapies such as hormones and external radiation are insignificant. In patients who experience biochemical failure it appears that PSA doubling time is an important predictor of survival. PMID- 18405744 TI - Commentary on prostate brachytherapy for localized prostate cancer. PMID- 18405745 TI - Incidence and location of prostate and urothelial carcinoma in prostates from cystoprostatectomies: implications for possible apical sparing surgery. AB - PURPOSE: Prostatic carcinoma (Pca) at cystoprostatectomy is usually an incidental finding with the majority thought to be clinically insignificant. Most studies have not specifically addressed the location of Pca or the incidence and location of in situ or invasive urothelial carcinoma (Uca) in prostates of cystoprostatectomy specimens. The frequency of involvement of the apex with these processes has clinical implications. Specifically urinary continence following orthotopic diversion may be enhanced by prostate apical sparing. In this study the pathological features of Pca and Uca, and the frequency of apical involvement were investigated in prostates from cystoprostatectomy specimens. MATERIALS AND METHODS: Whole mounted prostates from 121 consecutive cystoprostatectomy specimens were analyzed. Pca location, tumor volume, grade, stage, surgical margin and pelvic lymph node status of Pcas were assessed. Clinically insignificant Pcas had a volume of less than 0.5 cc without Gleason pattern 4, extracapsular extension, seminal vesicle invasion, lymph node involvement or positive surgical margins. Prostate involvement by Uca or urothelial carcinoma in situ (CIS)/severe dysplasia and its location were assessed. RESULTS: Of 121 prostates 50 (41%) had unsuspected Pca, of which 24 (48%) were clinically significant. Of Pcas 30 of 50 (60%) involved the apex, including 19 of 24 (79%) that were significant and 11 of 26 (42%) that were insignificant. Of 121 prostates 58 (48%) had Uca involving the prostatic stroma, noninvasive Uca or urothelial CIS/severe dysplasia in the prostatic urethra or periurethral ducts, of which 19 (33%) had apical involvement. Overall only 32 of 121 patients (26%) had no Pca or prostate Uca/CIS and only 45 (37%) had no clinically significant Pca or Uca/CIS in the prostate. However, 74 of the 121 patients (61%) had no prostatic apical involvement by Pca or Uca/CIS and 85 (70%) had no apical involvement by clinically significant Pca or Uca/CIS. Patients with prostatic apical involvement by invasive or in situ Uca uniformly had involvement of more proximal (toward the base) portions of the prostate. CONCLUSIONS: The majority of prostates from cystoprostatectomies had no involvement of the prostatic apex by Uca or clinically significant Pca. Hence, most patients may be candidates for prostate apical sparing. However, involvement of the apex by Uca in any patient raises concern about procedures that leave portions of the prostate urethra after cystectomy in an effort to improve continence. In candidates for orthotopic neobladder reconstruction removing all of the prostatic urethra and sparing the remainder of the prostatic apex may allow improved preservation of urinary continence with an acceptable low risk of clinical Pca progression. Whether future strategies for preoperative exclusion of apical Pca and intraoperative assessment of more proximal prostate to help exclude apical urothelial disease may identify patients suitable for prostatic apical sparing remains to be determined. The impact on functional outcomes and cancer control also require additional study. PMID- 18405746 TI - Commentary on prostate and urothelial carcinoma in cystoprostatectomy specimens. PMID- 18405747 TI - Clinical outcomes after sexuality preserving cystectomy and neobladder (prostate sparing cystectomy) in 44 patients. AB - PURPOSE: We describe the functional outcome on erectile function, continence and voiding, and local and distant cancer recurrence rates in 44 patients after sexuality preserving cystectomy and neobladder (prostate sparing cystectomy). MATERIALS AND METHODS: A total of 44 males underwent cystectomy with preservation of the prostate, seminal vesicles and vasa deferentia, after which a Studer type neobladder was anastomosed to the prostate. Oncological outcome (disease specific survival, distant and local recurrence rates) and functional results (continence, voiding, erectile function) were determined. RESULTS: At a median followup of 42 months, 13 (30%) patients died of cancer. All 13 experienced widespread disease, which was combined with a pelvic recurrence (pelvic recurrence rate 6.9%) in 3. The 3-year survival according to pathological stage was 86% for pT 2N0 or lower, 63% pT3N0 and 39% for node positive tumors (anyT Npos). Prostate cancer was diagnosed in 1 patient 5 years after treatment, and recurrent carcinoma in situ in the prostatic urethra in another patient. Complete daytime and nighttime continence was achieved in 95.3% and 74.4%, respectively. Incontinence during day and night could be managed by 1 pad per day/night in 4.7% and 20.9%, respectively, while 4.7% needed more than 1 pad per night. Erectile function could be determined in 40 patients, and potency was maintained in 77.5%, impaired in 12.5% and absent in 10%. CONCLUSIONS: Functional results with regard to erectile function and urinary continence after prostate sparing cystectomy are good. Oncological results have been promising, but need to be confirmed after longer followup and in larger trials. PMID- 18405748 TI - Commentary on clinical outcomes of sexuality preserving cystectomy and neobladder. PMID- 18405749 TI - 5-year urinary and sexual outcomes after radical prostatectomy: results from the Prostate Cancer Outcomes Study. AB - PURPOSE: Prior studies of postoperative outcomes following radical prostatectomy have been limited by selection bias and short-term followup. In this study we assessed temporal changes in urinary and sexual function up to 5 years following radical prostatectomy in a population based cohort. MATERIALS AND METHODS: A sample of 1,288 men with localized prostate cancer who underwent radical prostatectomy and completed a baseline survey within 6 to 12 months of diagnosis were included in the analysis. Two and 5-year functional and quality of life data were collected, as was information on the use of erectile aids. Temporal functional changes and potentially confounding or modifying factors were assessed using longitudinal regression models. RESULTS: Of these men 14% reported frequent urinary leakage or no urinary control 60 months after diagnosis, which was slightly higher than the 10% reporting incontinence at 24 months (p = 0.007). At 60 months 28% of the men had erections firm enough for intercourse compared with 22% at 24 months (p = 0.003). Sildenafil was the most commonly used erectile aid (43% ever used) and 45% of users reported that it helped "somewhat" or "a lot." CONCLUSIONS: Urinary and sexual dysfunction were common 5 years following radical prostatectomy in this large, community based cohort of prostate cancer survivors. While a small minority of subjects experienced changes in urinary or sexual function between years 2 and 5 after prostatectomy, functional outcomes remained relatively stable in the majority of participants. PMID- 18405750 TI - Commentary on urinary and sexual outcomes after radical prostatectomy. PMID- 18405751 TI - Do margins matter? The prognostic significance of positive surgical margins in radical prostatectomy specimens. AB - PURPOSE: The prognostic significance of positive surgical margins (PSM) in radical prostatectomy (RP) specimens remains unclear. While most studies have concluded that a PSM is an independent adverse prognostic factor, others report that surgical margin status has no effect on prognosis. One reason for these discordant conclusions is the variable number of patients with a PSM who receive adjuvant therapy and the differing statistical methods used to account for the effects of the time course of adjuvant treatment on recurrence. We evaluated the prognostic significance of PSMs using multiple methods of analysis accounting for patients who received adjuvant therapy. MATERIALS AND METHODS: We analyzed 1,389 consecutive patients with clinical stage T1-3 prostate cancer treated with RP by 2 surgeons from 1983 to 2000. Of 179 patients with a PSM, 37 received adjuvant therapy (AT), 29 radiation therapy and 8 received hormonal therapy. Because the method used to account for men receiving AT can affect the outcome of the analysis, data were analyzed by the Cox proportional hazards technique accounting for patients receiving AT using 5 methods: 1) exclusion, 2) inclusion (AT ignored), 3) censoring at time of AT, 4) failing at time of AT and 5) considering AT as a time dependent covariate. RESULTS: Overall 179 patients (12.9%) had a PSM, including 6.8% of 847 patients with pT2 and 23% of 522 patients with pT3 disease. A PSM was a significant predictor of cancer recurrence when analyzed using methods 1, 3, 4 and 5 (p=0.005, p=0.014, p=0.0005, p=0.002, respectively). However, it was not a predictor of recurrence using method 2 in which AT was ignored (p=0.283). Using method 5 multivariate analysis demonstrated that a PSM (p=0.002) was an independent predictor of 10-year progression-free probability (PFP) along with Gleason score (p=0.0005), extracapsular extension (p=0.0005), seminal vesicle invasion (p <0.0005), positive lymph nodes (p <0.0005) and preoperative serum prostate specific antigen (p <0.0001). Using method 5 the 10 year PFP was 58% +/- 12% and 81% +/- 3% for patients with and without a PSM, respectively (p <0.00005). The relative risk of recurrence in men with a PSM using method 5 was 1.52 (95% confidence interval 1.06-2.16). CONCLUSIONS: We confirm that a PSM has a significant adverse impact on PFP after RP in multivariate analysis using multiple statistical methods to account for patients who received AT. While prostate cancer screening strategies have resulted in a majority of men having organ confined disease at RP, surgeons should continue to strive to reduce the rate of positive surgical margins to improve cancer control outcomes. PMID- 18405752 TI - Commentary on significance of positive surgical margins. PMID- 18405753 TI - Early versus delayed hormonal therapy for prostate specific antigen only recurrence of prostate cancer after radical prostatectomy. AB - PURPOSE: Hormonal therapy (HT) is the current mainstay of systemic treatment for prostate specific antigen (PSA) only recurrence (PSAR), however, there is virtually no published literature comparing HT to observation in the clinical setting. The goal of this study was to examine the Department of Defense Center for Prostate Disease Research observational database to compare clinical outcomes in men who experienced PSAR after radical prostatectomy by early versus delayed use of HT and by a risk stratified approach. MATERIALS AND METHODS: Of 5,382 men in the database who underwent primary radical prostatectomy (RP), 4,967 patients were treated in the PSA-era between 1988 and December 2002. Of those patients 1,352 men who had PSAR (PSA after surgery greater than 0.2 ng/ml) and had postoperative followup greater than 6 months were used as the study cohort. These patients were further divided into an early HT group in which patients (355) received HT after PSA only recurrence but before clinical metastasis and a late HT group for patients (997) who received no HT before clinical metastasis or by current followup. The primary end point was the development of clinical metastases. Of the 1,352 patients with PSAR clinical metastases developed in 103 (7.6%). Patients were also stratified by surgical Gleason sum, PSA doubling time and timing of recurrence. Univariate and multivariate Cox proportional hazard models were used to evaluate the effect of early and late HT on clinical outcome. RESULTS: Early HT was associated with delayed clinical metastasis in patients with a pathological Gleason sum greater than 7 or PSA doubling time of 12 months or less (Hazards ratio = 2.12, p = 0.01). However, in the overall cohort early HT did not impact clinical metastases. Race, age at RP and PSA at diagnosis had no effect on metastasis-free survival (p >0.05). CONCLUSIONS: The retrospective observational multicenter database analysis demonstrated that early HT administered for PSAR after prior RP was an independent predictor of delayed clinical metastases only for high-risk cases at the current followup. Further study with longer followup and randomized trials are needed to address this important issue. PMID- 18405754 TI - Commentary on association of clinical prostate and bladder cancers. PMID- 18405755 TI - Commentary on early hormonal therapy for PSA only recurrence after radical prostatectomy--success towards survival or antidote to anxiety? PMID- 18405756 TI - Mechanisms in prostatitis/chronic pelvic pain syndrome. AB - PURPOSE: We reviewed the current literature on mechanisms involved in the pathogenesis of prostatitis/chronic pelvic pain syndrome (CPPS). MATERIALS AND METHODS: A literature review for the years 1966 to 2003 was performed using the MEDLINE database of the United States National Library of Medicine. RESULTS: National Institutes of Health categories I and II prostatitis result from identifiable prostatic infections, whereas patients with category IV are asymptomatic. The majority of symptomatic cases are category III or chronic prostatitis (CP)/CPPS. The etiology of CP/CPPS is unknown. The traditional marker of inflammation, namely white blood cells in prostatic fluids, does not correlate with the predominant symptom of pelvic pain. An imbalance toward increased proinflammatory and decreased anti-inflammatory cytokines has been implicated and a few studies have shown some correlation of this with pelvic pain. The imbalance in some men may result from polymorphisms at the cytokine loci. An autoimmune process may be involved and experimental evidence indicates that this can be under hormonal influence. Recent findings include possible defects in the androgen receptor. The prostate may not even be the source of the symptoms. Pelvic pain also correlates with the neurotrophin nerve growth factor implicated in neurogenic inflammation and central sensitization. Finally, psychological stress may produce measurable biochemical changes and influence the other processes. The role of normal prostatic bacterial flora in inciting the inflammatory response has also been reconsidered. CONCLUSIONS: The symptoms of CP/CPPS appear to result from an interplay between psychological factors and dysfunction in the immune, neurological and endocrine systems. PMID- 18405757 TI - Commentary on the mechanisms in prostatitis/chronic pelvic pain syndrome. PMID- 18405758 TI - Prospective randomized trial comparing shock wave lithotripsy and ureteroscopy for lower pole caliceal calculi 1 cm or less. AB - PURPOSE: The optimal management of lower pole renal calculi is controversial. We compared shock wave lithotripsy (SWL) and ureteroscopy (URS) for the treatment of patients with small lower pole stones in a prospective, randomized, multicenter trial. MATERIALS AND METHODS: A total of 78 patients with 1 cm or less isolated lower pole stones were randomized to SWL or URS. The primary outcome measure was stone-free rate on noncontrast computerized tomography at 3 months. Secondary outcome parameters were length of stay, complication rates, need for secondary procedures and patient derived quality of life measures. RESULTS: A total of 67 patients randomized to SWL (32) or URS (35) completed treatment. The 2 groups were comparable with respect to age, sex, body mass index, side treated and stone surface area. Operative time was significantly shorter for SWL than URS (66 vs 90 minutes). At 3 months of followup 26 and 32 patients who underwent SWL and URS had radiographic followup that demonstrated a stone-free rate of 35% and 50%, respectively (p not significant). Intraoperative complications occurred in 1 SWL case (unable to target stone) and in 7 URS cases (failed access in 5 and perforation in 2), while postoperative complications occurred in 7 SWL and 7 URS cases. Patient derived quality of life measures favored SWL. CONCLUSIONS: This study failed to demonstrate a statistically significant difference in stone-free rates between SWL and URS for the treatment of small lower pole renal calculi. However, SWL was associated with greater patient acceptance and shorter convalescence. PMID- 18405759 TI - Bladder cancer risk following primary and adjuvant external beam radiation for prostate cancer. AB - PURPOSE: Increased rates of secondary bladder malignancies have been reported after external beam radiation therapy (EBRT) for gynecological malignancies with relative risks of 2 to 4. This study was designed to determine if there was an increase in bladder cancer after EBRT for prostate cancer. MATERIALS AND METHODS: We retrospectively reviewed the Mayo Clinic Cancer Registry for patients who received EBRT for prostate cancer (1980 to 1998). Patients diagnosed with bladder cancer were identified. Comparative incidence rates were obtained from the national Surveillance, Epidemiology and End Results database. Subset analysis included patients treated with adjuvant radiation and those residing locally. Medical histories of patients with bladder cancer were reviewed. RESULTS: A total of 1,743 patients received EBRT for prostate cancer at our institution. In more than 12,353 man-years of followup no increase in bladder cancer risk was encountered. Subset analysis of men who received adjuvant radiation demonstrated that the relative risk of bladder cancer was increased but was not statistically significant. When the analysis was restricted to patients residing in the local area, the number of patients in whom subsequent bladder cancer developed was similar to Surveillance, Epidemiology and End Results rates. However, in the adjuvant radiation subset there was a statistically significant increase in subsequent bladder cancer. Patients in whom bladder cancer develops after EBRT often present with low grade disease but many have recurrence and progression. CONCLUSIONS: This retrospective review suggests there is not evidence of increased risk of bladder cancer after radiation therapy, assuming unbiased followup and complete ascertainment of cases. The natural history of bladder cancer in this population does not seem to be altered by a history of radiation. PMID- 18405760 TI - Commentary on shock wave lithotripsy versus ureteroscopy for lower pole caliceal calculi. PMID- 18405761 TI - Urologic diseases in america project: benign prostatic hyperplasia. AB - PURPOSE: Benign prostatic hyperplasia (BPH), the most common benign neoplasm in American men, is a chronic condition that is associated with progressive lower urinary tract symptoms and affects almost 3 of 4 men during the seventh decade of life. Approximately 6.5 million of the 27 million white men who are 50 to 79 years old in the United States in 2000 were estimated to meet the criteria for discussing treatment. MATERIALS AND METHODS: The analytical methods used to generate these results have been described previously. RESULTS: In 2000 approximately 4.5 million visits were made to physician offices to for a primary diagnosis of BPH and almost 8 million visits were made with a primary or secondary diagnosis of BPH. In the same year approximately 87,400 prostatectomies for BPH were performed in inpatients in nonfederal hospitals in the United States. While the number of outpatient visits for BPH increased consistently during the 1990s, there was a dramatic decrease in the use of transurethral prostatectomy, inpatient hospitalization and length of hospital stay for this condition. These trends reflect the changing face of medical management for BPH, ie increasing use of pharmacological agents and minimally invasive therapies. In 2000 the direct cost of BPH treatment was estimated to be $1.1 billion exclusive of outpatient pharmaceuticals. CONCLUSIONS: Given the impact that BPH has on quality of life and health care cost in millions of American men, additional research into risk factors, diagnostic and therapeutic resource use, and effectiveness and cost benefit of therapies are warranted. PMID- 18405762 TI - Commentary on BPH and public health--have we lost the forest through the trees? PMID- 18405763 TI - Tolterodine extended release attenuates lower urinary tract symptoms in men with benign prostatic hyperplasia. AB - PURPOSE: In this open label, prospective study we determined the efficacy and tolerability of tolterodine extended release (ER) in men with benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS) in whom previous alpha blocker therapy had failed. MATERIALS AND METHODS: A total of 43 consecutive men with BPH and LUTS in whom a mean of 5.7 months of alpha-blocker therapy had failed due to adverse events (11) or a lack of efficacy (32) received tolterodine ER (4 mg daily) for 6 months. Primary efficacy end points were American Urological Association symptom score, and mean daytime and nighttime micturition frequency. Secondary end points were the peak urinary flow rate, post-void residual volume, the incidence of urinary retention, total score on the erectile function domain of the International Index of Erectile Function and adverse events. RESULTS: A total of 39 men (91%) with a mean age of 61 years completed the 6-month trial. Mean 24-hour micturition frequency decreased from 9.8 to 6.3 voids and nocturia decreased from 4.1 to 2.9 episodes nightly. Significant changes in mean American Urological Association symptom scores (-6.1), the peak urinary flow rate (1.9 ml per second) and post-void residual volume (-22 ml) were also observed. Of the men 27 (63%) were potent at baseline and 29 (67%) were potent after 6 months of tolterodine ER treatment. Mean International Index of Erectile Function erectile function domain scores increased (6.9). Four men (9%) discontinued therapy because of intolerable dry mouth. There were no reports of urinary retention. CONCLUSIONS: Treatment with tolterodine ER in men with BPH and LUTS may be a reasonable therapeutic option as initial therapy or after failed treatment with alpha-blockers. PMID- 18405764 TI - Commentary on tolterodine for men with benign prostatic hyperplasia and lower urinary tract symptoms. PMID- 18405765 TI - Holmium laser enucleation versus transurethral resection of the prostate: results from a 2-center prospective randomized trial in patients with obstructive benign prostatic hyperplasia. AB - PURPOSE: To our knowledge we report the first multicenter, prospective, randomized study comparing holmium laser enucleation (HoLEP) and transurethral prostate resection (TURP) for obstructive benign prostatic hyperplasia. MATERIALS AND METHODS: From January to October 2002, 100 consecutive patients with symptomatic obstructive benign prostatic hyperplasia were randomized at 2 centers to surgical treatment with HoLEP (52 in group 1) or TURP (48 in group 2). Patients in the 2 groups were preoperatively assessed by scoring subjective symptoms questionnaires. Preoperative and perioperative parameters were also evaluated, the latter at 1, 6 and 12 months of followup. RESULTS: At baseline all patients had obstruction (Schafer grade greater than 2). At the 1, 6 and 12-month followups no statistically significant differences were observed between the 2 groups in terms of urodynamic findings and subjective symptom scoring. In the HoLEP group mean total time in the operating room +/- SD was significantly longer than for TURP (74 +/- 19.5 vs 57 +/- 15 minutes, p <0.05), while catheterization time (31 +/- 13 vs 57.78 +/- 17.5 minutes, p <0.001 and hospital stay (59 +/- 19.9 vs 85.8 +/- 18.9 hours, p <0.001) were significantly shorter in the HoLEP group. Transient stress and urge incontinence were more common in the HoLEP group, although at the 12-month followup results were comparable. The overall complication rate was comparable in the 2 groups. Erectile function was also maintained in the followup period from baseline in each group, as expected. CONCLUSIONS: HoLEP and TURP were equally effective for relieving obstruction and lower urinary tract symptoms. HoLEP was associated with shorter catheterization time and hospital stay. At 1 year of followup complications were similar in the 2 groups. PMID- 18405766 TI - Commentary on holmium laser enucleation versus TURP. PMID- 18405767 TI - Prospective, randomized, crossover comparison of sublingual apomorphine (3 mg) with oral sildenafil (50 mg) for male erectile dysfunction. AB - PURPOSE: We established the efficacy and safety of sublingual apomorphine compared with oral sildenafil in comparable groups of patients with erectile dysfunction (ED). MATERIALS AND METHODS: This prospective, randomized, crossover study included 77 heterosexual men with ED of various etiologies and severities. A total of 62 men were randomized but only 34 were evaluable for efficacy and tolerability. The study started with a run-in period of 2 to 4 weeks. The first 4 weeks of treatment were followed by a washout period of 4 weeks, after which patients changed to the alternate treatment for an additional 4-week period. The sequence of the 2 treatments was established by a randomization list in blocks in closed packets. The primary efficacy end point was the percent of attempts resulting in erection firm enough for intercourse. Additional variables were the percent of attempts resulting in intercourse and improvement in ED, as evaluated by the erectile function domain score of the International Index of Erectile Function questionnaire. RESULTS: Sildenafil was significantly more effective than apomorphine in regard to the percent of attempts resulting in erection firm enough for intercourse (85% vs 44%, p <0.0001) and actually resulting in intercourse (81% vs 43%, p <0.0001) as well as erectile function evaluated by the erectile function domain score of the International Index of Erectile Function (p <0.001). The incidence of adverse events was not significantly different for the 2 drugs. Although the number of patients was small, this study had strong statistical power due to the striking difference in results. CONCLUSIONS: Sildenafil was significantly more effective than apomorphine for ED. No statistical difference in adverse events was noted. PMID- 18405768 TI - Commentary on apomorphine versus sildenafil for erectile dysfunction. PMID- 18405769 TI - Randomized study of testosterone gel as adjunctive therapy to sildenafil in hypogonadal men with erectile dysfunction who do not respond to sildenafil alone. AB - PURPOSE: We compare the efficacy of testosterone gel (T-gel) versus placebo as adjunctive therapy to sildenafil in hypogonadal men with erectile dysfunction who do not respond to sildenafil alone. MATERIALS AND METHODS: A randomized, placebo controlled, double-blind, parallel group, multicenter study was performed. A total of 75 hypogonadal men (18 to 80 years old, morning serum total testosterone 400 ng/dl or less) with confirmed lack of response to sildenafil monotherapy were randomized (1:1) to receive a daily dose of 1% T-gel or 5 gm placebo gel as adjunctive therapy to 100 mg sildenafil during a 12-week period. Subjects were evaluated for sexual function, primarily based on the International Index of Erectile Function (IIEF), quality of life and serum testosterone levels at baseline and weeks 4, 8 and 12. RESULTS: Testosterone treated subjects had greater improvement in erectile function compared to those who received placebo, reaching statistical significance at week 4 (4.4 vs 2.1, p = 0.029, 95.1% CI 0.3, 4.7). Similar trends were observed for improvements in orgasmic function, overall satisfaction, total IIEF score and percentage of IIEF responders. T-gel significantly (p < or = 0.004) increased total and free testosterone levels throughout the study, although no significant correlations were made between testosterone levels and the IIEF at end point. CONCLUSIONS: T-gel taken with sildenafil may be beneficial in improving erectile function in hypogonadal men with erectile dysfunction who are unresponsive to sildenafil alone. PMID- 18405770 TI - [Immunoablation and autologous hematopoietic stem cell transplantation (AHSCT) in multiple sclerosis]. AB - Numerous pathophysiological arguments supporting immunosuppression for multiple sclerosis have been collected during recent years. The relevance of intense immunosuppression, in terms of clinical benefit and early or late risk, remains a matter of discussion. Immunoablation followed by autologous hematopoietic stem cell transplantation (AHSCT) in multiple sclerosis uses intense immunosuppression, followed by reinjection of AHSC, a rescue procedure for the induced aplasia. This method targets disappearance of the immune disorder, and thus, in theory, the interruption of the disease course. Use of AHSCT to treat several types of autoimmune diseases has been performed with contrasted results. In multiple sclerosis, the experience has been gained over the past 10 years through short series of patients treated at a late stage of their disease. This article highlights the recent data of this particular treatment option in multiple sclerosis as well as the therapeutic aims that should be investigated in further trials. PMID- 18405771 TI - [Familial and sporadic hemiplegic migraine]. AB - Hemiplegic migraine (HM) is a rare variety of migraine with aura characterized by the presence of a motor weakness during the aura. Hemiplegic migraine has two main forms according to the familial history: patients with at least one first- or second-degree relative who has aura including motor weakness have familial hemiplegic migraine (FHM); patients without such familial history have sporadic hemiplegic migraine (SHM). The prevalence of HM is one in 10,000 with FHM and SHM being equally frequent. Typical HM attacks include a motor weakness that is always associated with other aura symptoms, the most frequent being sensory, visual and speech disorders. In addition, basilar-type symptoms occur in up to 70% of the patients. Severe attacks may occur in FHM as well as in SHM with prolonged hemiplegia, confusion, coma, fever and seizures. The clinical spectrum also includes permanent cerebellar signs (nystagmus, ataxia, dysarthria) and less frequently various types of seizures and intellectual deficiency. FHM is the only variety of the autosomal dominant migraine and all three know genes encode ion transporters. A genetic diagnosis is now possible by screening the three known genes involved in FHM (CACNA1A, ATP1A2 and SCNA1). Prognosis is usually good. Treatment is similar to approaches used in other varieties of migraine with aura, excepted for triptans that are contraindicated in MHF/MHS. Based on new pathophysiological insight, preventive treatments by various antiepileptic agents seem promising. PMID- 18405772 TI - [Primary brain stem hemorrhage: retrospective study of 25 cases]. AB - INTRODUCTION: We report a retrospective series of 25 cases of brain stem hemorrhage. METHODS: Cases of spontaneous hemorrhage of the brain stem which were observed from 1990 to 2000 in a department of neurology were reviewed. Etiological factors, CT scan at admission, clinical signs and the course of the disease were analyzed retrospectively. RESULTS: There were 25 patients, 14 male and 11 female aged from 24 to 91. Fifteen hematomas were related to hypertension, four to coagulation disorders and two to a vascular malformation. The hemorrhage was located in the pons in 22 cases and in the midbrain in three cases. The death rate directly related to the hemorrhage was 14/25 (12 early and two delayed deaths). Prognosis factors were the size of the hemorrhage, a ventricular bleeding, disorders of consciousness and pupillary abnormalities on admission, the need for mechanical ventilation. CONCLUSION: In brain stem hemorrhage, the size of the hematoma is a more important prognosis factor than age or etiological factors. PMID- 18405773 TI - [Management of isolated optic neuritis in France: survey of neurologists and ophthalmologists]. AB - BACKGROUND: Acute isolated optic neuritis is often the first manifestation of multiple sclerosis (MS). Despite the results of several clinical trials its management remains controversial. With the advent of new disease-modifying agents for the treatment of MS, management of isolated optic neuritis has become more complicated. The goal of this study was to evaluate the current clinical practice of French ophthalmologists and neurologists in the management of acute isolated optic neuritis, and to evaluate the impact of recently published randomized clinical trials on their practice. METHODS: A survey, including 24 questions on the diagnosis and treatment of acute isolated optic neuritis was sent to all neurologists and to a sample of ophthalmologists in France. RESULTS: The responses of 655 neurologists and 141 ophthalmologists were analyzed. This study shows mostly that patients initially present more frequently to ophthalmologists, and are subsequently referred to neurologists. Most optic neuritis patients undergo a brain MRI and a lumbar puncture. Although most patients receive high dose intravenous steroids, up to 15% of neurologists and 21% of ophthalmologists still recommend oral prednisone (1 mg/kg per day). Steroids are often prescribed for the wrong reason, including to improve final visual acuity or decrease the risk of MS. Disease modifying agents are sometimes prescribed outside of the official French recommendations. CONCLUSION: The evidence-based guidelines are only partially followed by practitioners managing patients with acute optic neuritis. PMID- 18405774 TI - [Alexithymia and links with depression and anxiety in multiple sclerosis]. AB - OBJECTIVES: This study evaluates the prevalence of alexithymia in multiple sclerosis and examines the links between alexithymia, depression and anxiety. METHOD: Sixty-one subjects aged between 18 and 60 years and suffering from multiple sclerosis took part in the study. The psychological assessment consisted of an interview with a psychologist and three questionnaires: the Toronto Alexithymia Scale (TAS), the State-Trait Anxiety Inventory (STAI), the Beck Depression Inventory (BDI). RESULTS: The prevalence of alexithymia was 42.5%, 34.4% for depression and 44.3% for anxiety (high and moderate level). The alexithymic subjects were more depressed and anxious. Results indicated positive correlations between anxiety (state and trait), depression and alexithymia scores. The various dimensions of alexithymia were found to be diversely correlated with anxiety and depression. CONCLUSIONS: Our results point out the importance of anxiety in multiple sclerosis and the specificity of alexithymia. PMID- 18405775 TI - [Epileptic seizure and migraine visual aura: revisiting migralepsy]. AB - INTRODUCTION: The term of "migralepsy" has been proposed to define migraine triggered epileptic seizures. Although already reported in the literature for more than fifty years, a number of observations remain debatable because of possible confusion between migraine and epileptic seizure clinical manifestations, including hemifield visual hallucinations, digestive signs and severe headache. OBSERVATION: We report on the case of a young patient suffering from both diseases, in whom a visual aura preceded either migraine attacks or epileptic generalized tonic-clonic seizure. Subtle modification in the primitive visual hallucination, which suddenly contained colored figures and was accompanied by fear before a prolonged loss of contact, suggested a continuum between migraine aura and epileptic seizure in this patient. Brain MRI was normal and EEG showed some sharp waves in the right posterior area. CONCLUSION: The presence of a neurophysiological continuum between migrainous aura and epileptic seizure is supported by this observation of "migralepsy". Recent findings from genetic and epidemiological studies further support this link. PMID- 18405776 TI - [Neurosyphilis revealed by a multiple cranial neuropathy: magnetic resonance imaging findings]. AB - INTRODUCTION: We report the magnetic resonance imaging (MRI) findings in a case of neurosyphilis revealed by the involvement of two cranial nerves. CASE REPORT: A 41-year-old man developed a right cochleovestibular and left trigeminal neuropathy, associated with high serum titers of VDRL and TPHA, high titers of TPHA in the cerebrospinal fluid (CSF) and several CSF oligoclonal IgG bands. On MRI, hypertrophy and gadolinium contrast enhancement of these cranial nerves were associated with several supratentorial cortical nodules surrounded by marked cerebral edema, corresponding to syphilitic gummas. One of these cortical nodules was biopsied. Microscopic examination showed lesions of meningoencephalitis with necrosis and granulomatous vasculitis. After penicillin therapy, the serum VDRL titers and the MRI abnormalities disappeared, a partial clinical recovery was observed and a significant reduction of the serum TPHA titers was found. DISCUSSION: Such MRI abnormalities are not specific and can be observed in various tumoral, auto-immune and infectious diseases. They can also mimic neurofibromatosis type II. Cranial nerve involvements in neurosyphilis can result from nerve inflammation in basal meningitis, nerve ischemia in meningovasculitis or from compression by an adjacent gumma. In our case, the cranial neuropathy was related to a mixed meningovascular and parenchymatous form of neurosyphilis. PMID- 18405777 TI - [Visual auras in idiopathic generalized epilepsy]. AB - Visual auras are reported to be uncommon or rare in patients with idiopathic generalized epilepsy. We describe three patients who experienced visual auras just before generalized tonic-clonic seizures. This study indicates that in idiopathic generalized epilepsy, visual auras manifest as flashes, lighting or the impression of seeing the sun. Unlike auras in occipital lobe epilepsy, they are very brief. PMID- 18405778 TI - [Superficial hemosiderosis of the central nervous system improved by corticosteroids]. AB - INTRODUCTION: Superficial siderosis is of the Central Nervous System (CNS) is an uncommon and often disabling disorder. There is no evidence that any treatment, including removal of an identified source of bleeding, affects disease progression. OBSERVATION: We report the case of a 49-year-old woman exhibiting progressive and various neurological disorders associating chorea, pyramidal syndrome, cerebellar ataxia, cognitive disorders and cranial nerve deficits. She had a prior history of right occipital arterioveinous malformation (AVM) revealed four years before by ventricular hemorrhage. The AVM was treated by radiosurgery. Because of a pronounced progression of the symptoms, treatment with steroid therapy was initiated before the diagnosis of siderosis of the central nervous system was asserted by magnetic resonance imaging (rim of hypo-intensity due to hemosiderin around the brainstem, the cerebellum and the spinal cord on T-2 weighted and gradient echo T-2* imaging) and cerebrospinal fluid (CSF) examination (high CSF levels of iron and ferritin). Over the next months the neurological condition improved under steroid therapy. CONCLUSION: Our observation is interesting because of the chorea movement disorders which are rarely reported in the disease and because of the improvement of the neurological condition after steroid therapy which is described in only another case in the literature. Steroid therapy could constitute a new track for the treatment of siderosis of CNS. PMID- 18405779 TI - [Searching for lost memory: memory loss and recovery mechanisms observed in a patient with pure retrograde amnesia]. AB - We describe a patient who developed acute reversible amnesia characterized by impaired recollection of past events with preserved anterograde memory. This occurred after a mild head trauma and in the context of occupational stress. Isolated retrograde amnesia affected autobiographical memory (with lost of identity lasting a few days) and semantic knowledge. Isolated retrograde amnesia subsided for nine months. The patient was no longer able to access to his memories, which did not seem completely lost. Some answers to tests concerning historical events were better than those given at random; his answers to a TV quiz were automatic or he exhibited temporal transfer phenomena (ecmnesia). Gradual, progressive, irregular and spontaneous or "flash" recovery, which was also facilitated by dreams, associations or contextual clues. The persistent and vivid familiarity of the retrieved memories are reported here together with the organization of long-term memory and the clinical and neuropsychological traits of functional isolated retrograde amnesia. PMID- 18405780 TI - [Isolated pulmonary hypertension and pergolide]. AB - Pergolide, an ergot-derived dopamine agonist prescribed since the late 1980's mainly in Parkinson's disease and restless leg syndrome has recognized fibrosis side effects, affecting the pleural, pericardial and retroperitoneal systems. Pergolide-induced valvulopathies were first reported in 2002. We present here the history of a patient developing an isolated pulmonary hypertension related to the intake of pergolide. The dyspnea related to the pulmonary hypertension as well as the echocardiographic abnormalities improved after treatment replacement. Valvulopathies and pulmonary hypertension were previously described under appetite-suppressant drugs after years of clinical use, in a similar way. PMID- 18405783 TI - Clinical and patient-reported outcomes in clinical trials of abatacept in the treatment of rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic joint inflammation, which if left untreated leads to progressive disability and joint destruction. A combination of antiinflammatory agents, steroids, disease-modifying antirheumatic drugs, and biological agents are used to treat RA. Beyond the use of conventional measures of disease activity, such as American College of Rheumatology (ACR) response rates, the importance of patient reported outcomes (PROs) in assessing therapeutic benefits is gaining increasing emphasis in clinical trials of RA and other chronic illnesses. Clinical trials testing new RA therapeutics generally include health-related quality of life (HRQoL) measures and assessments of function and disability. Abatacept, a costimulation modulator that selectively targets the activation of T cells and downregulates the immune response, has been approved by the US Food and Drug Administration for the treatment of RA, with or without methotrexate. OBJECTIVE: The aim of this review was to summarize the clinical outcomes and PROs in published trials of abatacept. METHODS: A literature search was performed using the MEDLINE, EMBASE, and BIOSIS databases (restricted to articles posted between January 2000 and September 2007) with the search terms CTLA-4Ig, abatacept, and Orencia to identify published trials of abatacept. Primary clinical trial publications in patients with RA were selected. The ACR response and PROs data presented in the identified publications are summarized in this review. RESULTS: Our search identified 6 studies that met our selection criteria, which included 1 Phase IIa study, 2 Phase IIb studies, and 3 Phase III studies. The Phase IIa study found that abatacept was more effective than placebo and that physical function improved in treated patients compared with placebo. The 2 Phase IIb studies in 339 patients with RA previously treated with methotrexate found statistically significant improvements in HRQoL with abatacept at 6 months and 1 year. Similar findings were noted in the published Phase III trials. Across clinical trials, abatacept has been associated with clinically meaningful and statistically significant improvements in conventional measures of disease activity, HRQoL, and physical function. CONCLUSIONS: These 6 published trials found that abatacept was associated with significant improvements in both conventional measures of disease activity and PROs. Continued assessment of these outcomes will be required to further support the findings of the Phase II and III abatacept clinical trial literature reviewed here. PMID- 18405784 TI - Significance of a decline in bone mineral density while receiving oral bisphosphonate treatment. AB - BACKGROUND: Oral bisphosphonates are routinely prescribed for the treatment of postmenopausal osteoporosis. In clinical trials, oral bisphosphonates have been found to increase bone mineral density (BMD) and decrease fracture risk in the majority of the treated population. However, in both clinical trials and clinical practice, not all patients experience significant increases in BMD. In clinical trials, nonresponse is often defined as a BMD change of or=1 week). Venous blood samples were obtained for quantification of plasma fentanyl concentrations before and at 1, 3, 5, 7, 9, 12, 15, 25, 40, 60, 90, 120, and 180 minutes after administration. Pain scores (on an 11-point numeric rating scale) were recorded before and at 15, 30, 45, 60, 75, 90, 105, 120, 150, 180, 210, and 240 minutes. Patients indicated the times at which they perceived meaningful pain relief (onset of action) and at which analgesia ended (duration of effect), after which they were able to use rescue medication (time to rescue medication use). RESULTS: A total of 24 patients were enrolled (in all, 47 extractions) (46% male; mean age, 24.1 years; 94% white, 6% Asian). Mean T(max) values were 12.8 and 6.0 minutes (P<0.001), times to onset of analgesia were 7 and 2 minutes (P<0.001), and durations of effect were 56 and 59 minutes after i.n. and i.v. administration (P=NS), respectively. Differences in the onsets and durations of analgesia after i.n. and i.v. administration of single doses were not significantly different, and neither was the difference in overall analgesia, with pain scores returning to near-predose values at statistically similar times after dosing. Duration of effect was directly related to i.n. fentanyl dose, with significantly less use of rescue medication after i.n. than after i.v. administration (P<0.005). The i.n. and i.v. formulations were both well tolerated, with similar numbers of nasally related adverse events recorded for both routes of administration. CONCLUSIONS: Onsets and durations of analgesia were not significantly different between single doses of i.n. and i.v. fentanyl in these adults undergoing third-molar extraction. Both i.n. and i.v. administration were generally well tolerated. PMID- 18405787 TI - The effects of amlodipine and enalapril on renal function in adults with hypertension and nondiabetic nephropathies: a 3-year, randomized, multicenter, double-blind, placebo-controlled study. AB - BACKGROUND: Placebo-controlled trials have found that angiotensin-converting enzyme inhibitors (ACEIs) decrease proteinuria and slow the progression of nondiabetic nephropathies. However, head-to-head comparisons of ACEIs and calcium channel blockers (CCBs) have shown conflicting results. Indeed, a recent metaanalysis concluded that there is still uncertainty about the greater renoprotection seen with ACEIs or angiotensin II receptor blockers in nondiabetic patients with renal disease, particularly when using true glomerular filtration rate (GFR) as the primary outcome. OBJECTIVE: The objective of this 3-year, randomized, multicenter, double-blind, placebo-controlled study was to compare true GFR decline (measured by yearly 51Cr-EDTA blood clearance) in nondiabetic, nonnephrotic adult hypertensive patients with estimated creatinine clearance of 20 to 60 mL/min.1.73 m(2), when randomized to a CCB (amlodipine, 5-10 mg/d) or an ACEI (enalapril, 5-20 mg/d). METHODS: Patients (aged 18-80 years) entered a 4 week placebo run-in washout period and previous antihypertensive drugs were tapered off over 2 weeks. Add-on treatments were atenolol (50-100 mg/d), loop diuretics (furosemide, 20-500 mg/d or torsemide, 5-200 mg/d), alpha-blockers (prazosin, 2.5-5 mg/d or doxazosin, 1-16 mg/d), and centrally acting drugs (rilmenidine, 1-2 mg/d or methyldopa, 250-500 mg/d). The primary end point was true GFR measured by yearly (51)Cr-EDTA blood clearance. Secondary end points included a clinical composite of renal events and tolerability collected by a full clinical and laboratory evaluation at each study visit. Post hoc analyses for the change in GFR, proteinuria, and time to clinical events were also planned on baseline proteinuria subgroups (<1 and >or=1 g/d) before unblinding the database. RESULTS: Three hundred eighteen patients entered the run-in period and 263 patients (156 men/107 women; mean age, 58 years) were randomized to receive either amlodipine (5 mg/d, n=132) or enalapril (5 mg/d, n=131). Blood pressure declined from 165/102 mm Hg to 138/84 mm Hg and 138/85 mm Hg with amlodipine and enalapril, respectively (no between-group significance). Only 20.8% of the patients randomized to ACEI treatment received diuretics at the last observation. No statistically significant difference was found between amlodipine and enalapril in GFR decline (-4.92 and -3.98 mL/min.1.73 m(2), respectively, at last observation) and composite secondary end point after a median follow-up of 2.9 years, including in the subgroup of patients with proteinuria >1 g/d at baseline. Protein excretion rate decreased significantly from baseline in patients taking enalapril plus diuretics (median -270 mg/d; P<0.001) but not in patients taking amlodipine plus diuretics (-25 mg/d at last observation). CONCLUSION: In this cohort of nondiabetic, nonnephrotic hypertensive patients, no statistically significant difference in true GFR decline was found over 3 years between amlodipine-treated patients and enalapril-treated patients with main add-on treatment with ss-blockers, including in the subgroup of patients with proteinuria >1 g/d. PMID- 18405788 TI - Pharmacokinetic, pharmacodynamic, and tolerability profiles of the dipeptidyl peptidase-4 inhibitor alogliptin: a randomized, double-blind, placebo-controlled, multiple-dose study in adult patients with type 2 diabetes. AB - BACKGROUND: Alogliptin is a highly selective dipeptidyl peptidase-4 (DPP-4) inhibitor that is under development for the treatment of type 2 diabetes (T2D). OBJECTIVES: This study was conducted to evaluate the pharmacokinetic (PK), pharmacodynamic (PD), and tolerability profiles and explore the efficacy of multiple oral doses of alogliptin in patients with T2D. METHODS: In this randomized, double-blind, placebo-controlled, parallel-group study, patients with T2D between the ages of 18 and 75 years were assigned to receive a single oral dose of alogliptin 25, 100, or 400 mg or placebo (4:4:4:3 ratio) once daily for 14 days. PK profiles and plasma DPP-4 inhibition were assessed on days 1 and 14. Tolerability was monitored based on adverse events (AEs) and clinical assessments. Efficacy end points included 4-hour postprandial plasma glucose (PPG) and insulin concentrations, and fasting glycosylated hemoglobin (HbA(1c)), C-peptide, and fructosamine values. RESULTS: Of 56 enrolled patients (57% women; 93% white; mean age, 55.6 years; mean weight, 89.8 kg; mean body mass index, 31.7 kg/m(2)), 54 completed the study. On day 14, the median T(max) was ~1 hour and the mean t(1/2) was 12.5 to 21.1 hours across all alogliptin doses. Alogliptin was primarily excreted renally (mean fraction of drug excreted in urine from 0 to 72 hours after dosing, 60.8%-63.4%). On day 14, mean peak DPP-4 inhibition ranged from 94% to 99%, and mean inhibition at 24 hours after dosing ranged from 82% to 97% across all alogliptin doses. Significant decreases from baseline to day 14 were observed in mean 4-hour PPG after breakfast with alogliptin 25 mg (-32.5 mg/dL; P=0.008), 100 mg (-37.2; P=0.002), and 400 mg (-65.6 mg/dL; P<0.001) compared with placebo (+8.2 mg/dL). Significant decreases in mean 4-hour PPG were also observed for alogliptin 25, 100, and 400 mg compared with placebo after lunch (-15.8 mg/dL [P=0.030]; -29.2 mg/dL [P=0.002]; -27.1 mg/dL [P=0.009]; and +14.3 mg/dL, respectively) and after dinner (-21.9 mg/dL [P=0.017]; -39.7 mg/dL [P<0.001]; -35.3 mg/dL [P=0.003]; and +12.8 mg/dL). Significant decreases in mean HbA(1c) from baseline to day 15 were observed for alogliptin 25 mg (-0.22%; P=0.044), 100 mg (-0.40%; P<0.001), and 400 mg (-0.28%; P=0.018) compared with placebo (+0.05%). Significant decreases in mean fructosamine concentrations from baseline to day 15 were observed for alogliptin 100 mg (-25.6 micromol/L; P=0.001) and 400 mg (-19.9 micromol/L; P=0.010) compared with placebo (+15.0 micromol/L). No statistically significant changes were noted in mean 4-hour postprandial insulin or mean fasting C-peptide. No serious AEs were reported, and no patients discontinued the study because of an AE. The most commonly reported AEs for alogliptin 400 mg were headache in 6 of 16 patients (compared with 0/15 for alogliptin 25 mg, 1/14 for alogliptin 100 mg, and 3/11 for placebo), dizziness in 4 of 16 patients (compared with 1/15, 2/14, and 1/11, respectively), and constipation in 3 of 16 patients (compared with no patients in any other group). No other individual AE was reported by >2 patients receiving the 400-mg dose. Apart from dizziness, no individual AE was reported by >1 patient receiving either the 25- or 100-mg dose. CONCLUSIONS: In these adult patients with T2D, alogliptin inhibited plasma DPP-4 activity and significantly decreased PPG levels. The PK and PD profiles of multiple doses of alogliptin in this study supported use of a once-daily dosing regimen. Alogliptin was generally well tolerated, with no dose-limiting toxicity. PMID- 18405789 TI - Pharmacokinetics, pharmacodynamics, and tolerability of single increasing doses of the dipeptidyl peptidase-4 inhibitor alogliptin in healthy male subjects. AB - BACKGROUND: Alogliptin is a highly selective dipeptidyl peptidase-4 (DPP-4) inhibitor that is under development for the treatment of type 2 diabetes. OBJECTIVE: This study was conducted to characterize the pharmacokinetics, pharmacodynamics, and tolerability of single oral doses of alogliptin in healthy male subjects. METHODS: This was a randomized, double-blind, placebo-controlled study in which healthy, nonobese male subjects between the ages of 18 and 55 years were assigned to 1 of 6 cohorts: alogliptin 25, 50, 100, 200, 400, or 800 mg. One subject in each cohort received placebo. An ascending-dose strategy was used, in which each cohort received its assigned dose only after review of the safety data from the previous cohort. Blood and urine were collected over 72 hours after dosing for pharmacokinetic analysis and determination of plasma DPP-4 inhibition and active glucagon-like peptide -1(GLP-1) concentrations. RESULTS: Thirty-six subjects (66 per cohort) were enrolled and completed the study (29/36 [81% ] white; mean age, 26.6 years; mean weight, 76.0 kg). Alogliptin was rapidly absorbed (median T(max), 1-2 hours) and eliminated slowly (mean t(1/2), 12.4-21.4 hours), primarily via urinary excretion (mean fraction of drug excreted in urine from 0 to 72 hours after dosing, 60%-71%). C(max) and AUC(0-infinity) increased dose proportionally over the range from 25 to 100 mg. The metabolites M-I (N demethylated) and M-II (N-acetylated) accounted for <2% and <6%, respectively, of alogliptin concentrations in plasma and urine. Across alogliptin doses, mean peak DPP-4 inhibition ranged from 93% to 99%, and mean inhibition at 24 hours after dosing ranged from 74% to 97%. Exposure to active GLP-1 was 2- to 4-fold greater for all alogliptin doses compared with placebo; no dose response was apparent. Hypoglycemia (asymptomatic) was reported in 5 subjects (11 receiving alogliptin 50 mg, 2 receiving alogliptin 200 mg, 1 receiving alogliptin 400 mg, 1 receiving placebo). Other adverse events were reported in 1 subject each: dizziness (alogliptin 100 mg), syncope (alogliptin 200 mg), constipation (alogliptin 200 mg), viral infection (alogliptin 400 mg), hot flush (placebo), and nausea (placebo). CONCLUSION: In these healthy male subjects, alogliptin at single doses up to 800 mg inhibited plasma DPP-4 activity, increased active GLP-1, and was generally well tolerated, with no dose-limiting toxicity. PMID- 18405790 TI - Comparison of sequential and standard triple-drug regimen for Helicobacter pylori eradication: a 14-day, open-label, randomized, prospective, parallel-arm study in adult patients with nonulcer dyspepsia. AB - BACKGROUND: The eradication rates of Helicobacter pylori with standard treatments are decreasing worldwide. OBJECTIVES: The primary aim of this study was to compare the eradication success of a 14-day sequential regimen with proton pump inhibitor (PPI)-based triple treatment. The secondary objectives of the study were to evaluate the effect of gastritis score and smoking on eradication rates as well as evaluation of compliance and tolerability of both regimens. METHODS: Consecutive H pylori-positive patients with nonulcer dyspepsia were randomized into 1 of 2 groups in this 14-day, open-label, randomized, prospective, parallel arm study. An upper endoscopy with biopsy and (14)C-urea breath test ((14)C-UBT) were performed before enrollment. The first group was administered a sequential regimen consisting of pantoprazole 40 mg and amoxicillin 1 g for 7 days, followed by pantoprazole 40 mg, tetracycline 500 mg, and metronidazole 500 mg for the next 7 days. The second group was administered pantoprazole 40 mg, amoxicillin 1 g, and clarithromycin 500 mg (PAC group) for 14 days. All drugs were administered BID, with the exception of tetracycline, which was administered QID. Eradication was confirmed by (14)C-UBT 6 weeks after the end of the treatment. Histologic examination and (14)C-UBT were conducted by investigators blinded to the protocols. Patients were asked to report any adverse events (AEs) during the treatment period. RESULTS: Three hundred white patients were enrolled in the study and evenly randomized into the sequential treatment group (98 males and 52 females; mean age, 40.2 years) and the PAC group (86 males and 64 females; mean age, 41.2 years). A total of 274 patients completed the study per protocol (PP). Twenty-six patients discontinued: lost to follow-up (16), withdrawn due to AEs (9); and noncompliance (1). The intent-to-treat (ITT) and PP H pylori eradication rates were 72.6% and 80.1% in the sequential group, and 58% and 63% in the PAC group, respectively. The eradication rate was significantly higher in the sequential group compared with the PAC group in both the ITT and PP populations (P=0.01 and P=0.002, respectively). The eradication rates were higher in nonsmoking patients compared with smoking patients both in the sequential group (85.8% vs 70.5%) and the PAC group (67.7% vs 53.3%), but the results were not statistically significant when the groups were analyzed separately. Overall, 32 patients (10.7%) reported an AE. Treatment was discontinued in 9 patients because of serious AEs (sequential group--abdominal pain [2 patients], diarrhea [1], chest pain [1], and vaginal pruritus [1]; PAC group--nausea/vomiting [2], chest pain [1], and numbness [1]). There were no significant between-group differences in regard to compliance or AEs. Univariate analyses found no significant effect of sex, age, alcohol consumption, antacid usage, or gastritis score on the eradication rates. CONCLUSIONS: A 14-day sequential treatment regimen achieved a significantly higher eradication rate of H pylori compared with standard PPI based triple regimen in this small selected population. Large, double-blind, controlled studies are needed to confirm these results. PMID- 18405791 TI - The risk of myopathy associated with thiazolidinediones and statins in patients with type 2 diabetes: a nested case-control analysis. AB - BACKGROUND: The implementation of more aggressive goals for low-density lipoprotein cholesterol lowering in subjects with type 2 diabetes (T2D) and the expected increase in the use of statins is likely to increase the concomitant use of thiazolidinediones (TZDs) and statins. OBJECTIVE: This study evaluated whether concomitant use of TZDs and statins is associated with an increased risk of myopathic events in subjects with T2D. METHODS: This was a nested case-control study in subjects with T2D. Cases and controls were identified from a cohort of 125,394 subjects with T2D in the Integrated Healthcare Information Services database. Each case with a myopathic event (rhabdomyolysis, myositis, myopathy, or myalgia) was matched to up to 6 controls by age (+/-5 years), sex, calendar year of diagnosis of a myopathic event, and length of follow-up in the database. Incident cases of myopathy were identified using the following International Classification of Diseases, Ninth Revision codes: 359.x for myopathy, 728.88 for rhabdomyolysis, and 729.1 for unspecified myalgia and myositis. Prescription claims were used as a proxy for drug exposure. Five categories of exposure were employed: statins only, TZDs only, concomitant TZDs and statins, other antidiabetic agents only, and neither statins nor antidiabetic agents. Exposure to statins and/or TZDs within 90 days before the case index date was defined as recent exposure, and exposure at any time before the case index date was defined as ever exposure. Concomitant exposure to TZDs and statins, either recent or ever, was defined by an overlap of at least 30 days in the days supply of TZDs and statins during the exposure period. RESULTS: The 3696 cases of myopathy were matched with 21,871 controls. The adjusted odds ratio (OR) for myopathic events for ever exposure to concomitant TZDs and statins compared with statins alone was 1.03 (95% CI, 0.83-1.26). Compared with neither statins nor antidiabetic agents, ever use of statins alone was associated with an increased likelihood of myopathic events (adjusted OR=1.36; 95% CI, 1.12-1.64). The likelihood of myopathic events was not significantly different for TZDs compared with other antidiabetic agents. CONCLUSION: In this population of subjects with T2D, concomitant use of statins and TZDs was not associated with an increased risk of myopathic events beyond that conferred by statins alone. PMID- 18405792 TI - Assessing retinol stability in a hydroquinone 4%/retinol 0.3% cream in the presence of antioxidants and sunscreen under simulated-use conditions: a pilot study. AB - BACKGROUND: Retinol (ie,vitamin A) is commonly used in dermatology as an adjunct to treat rhytids, acne,and dyschromia. However, vitamin A and many of its derivatives have poor photostability and are unstable in the presence of oxygen. OBJECTIVE: We aimed to assess the stability of retinol under simulated patient application conditions in a commercially available hydroquinone 4% cream containing retinol 0.3%, avobenzone (ultraviolet-A sunscreen), octinoxate (ultraviolet-B sunscreen), vitamins C and E (antioxidants), and moisturizers. METHODS: One gram of the preparation was applied as a thin film to the inside base of 4 groups of four 100-mL wide-mouthed beakers, incubated in a 37+/-2 degrees C water bath. Each experimental group consisted of 4 beakers for assays at 0.5,1,2,and 4 hours. The samples were exposed to varying combinations of full spectrum light and headspace gas (air or inert nitrogen gas [N2 ]). Retinol content was assayed via high-pressure liquid chromatography using a 1:9 water:methanol solvent system. The control group (group 5) was not exposed to full-spectrum light or headspace gas but served for comparative purposes. RESULTS: On exposure to light and room air, retinol stability was 94.4% at 0.5 hour, 94.8% at 1 hour, 92.4% at 2 hours, and 91.5% at 4 hours. The retinol contained in the preparation was stable for >or=4 hours. Samples exposed to light and N 2 gas demonstrated 96.5% and 91.3% stability at 0.5 hour and 4 hours exposure times, respectively. Samples that were not exposed to light had a stability of 99.2% (group 3, exposed to air) and 96.9% (group 4, exposed to N(2)) of the initial retinol present after 4 hours. CONCLUSION: The retinol in the hydroquinone 4%/ retinol 0.3% cream with antioxidants and sunscreens underwent <10% degradation under simulated-use conditions, including exposure to UV light, oxygen, and body temperature. PMID- 18405793 TI - Cough induced by losartan with resolution after substitution with enalapril. AB - INTRODUCTION: Cough is an adverse event associated with the angiotensin converting enzyme (AA inhibitor drugs. ACE inhibitor-induced cough is believed to be related to the accumulation of bradykinin,substance P,and prostaglandins resulting from the inhibition of ACE.Angiotensin-receptor blockers (AARBs) do not have any effect on ACE and theoretically might not cause cough. Therefore, a proposed option in patients suffering with ACE inhibitor-induced cough is to try an ARB. However,this report describes the reverse: a case of losartan-induced cough th hat co om completely resolved after it was substituted with an ACE inhibitor, enalapril. CASE SUMMARY: A 23-year-old, nonsmoking white woman, weighing 73.55 kg, ACE inhibitor naive (before admission), presented to the emergency department at Imam Referral Hospital, Tehran, Iran,with hypertension,proteinuria, and hyperlipidemia. The patient was admitted to the nephrology ward. She was prescribed hydrochlorothiazide 12.55 mg/d, furosemide 20 mg BID, and simvastatin 20 mg/d. The patient had no respiratory illnesses. The patient experienced cough 3 days following the initiation of losartan treatment. The cough continued in this patient for the 2-week duration of losartan treatment; however, 1 week after substitution of losartan with enalapril (22.5 mg/d),the cough resolved completely. CONCLUSION: This report describes a young woman who developed cough while receiving losartan treatment,which resolved after substitution with the ACE inhibitor enalapril. PMID- 18405795 TI - Reauthorization of the State Children's Health Insurance Program: divergent ideologies or fiscal realities? PMID- 18405794 TI - Achieving glycosylated hemoglobin targets using the combination of repaglinide and metformin in type 2 diabetes: a reanalysis of earlier data in terms of current targets. AB - BACKGROUND: For patients with type 2 diabetes, the American Diabetes Association (ADA) and European Association for the Study of Diabetes (EASD) currently recommend a glycosylated hemoglobin (HbA(1c) ) target of <7%, and the British Medical Association (BMA) Quality and Outcomes Framework recommends an HbA(1c) target of >or=7.5%. OBJECTIVE: This letter presents a reanalysis of data from a previous study of the effect on glycemic control of adding repaglinide to metformin monotherapy in patients with type 2 diabetes to determine the proportion of patients achieving current ADA/EASD and BMA targets. METHODS: PubMed was searched using the terms repaglinide AND metformin AND HbA(1c) to identify published comparisons of monotherapy and combination therapy with these drugs in patients with type 2 diabetes. RESULTS: In the original analysis, which employed an HbA(1c) target of <7.1%, 59%of patients treated with metformin plus repaglinide achieved their glycemic target, compared with approximately 20% of patients treated with metformin or repaglinide alone. On reanalysis of the data according to the current ADA/EASD HbA(1c) target of <7%, 56% of patients receiving metformin and repaglinide achieved that goal,compared with 19%each in the groups treated with metformin or repaglinide monotherapy. On reanalysis of the data according to the BMA Quality and Outcomes Framework HbA(1c) target of >or=7.5%, 89% of patients receiving metformin and repaglinide achieved that goal, compared with 43%and 42% of patients receiving metformin and repaglinide monotherapy, respectively. CONCLUSION: Based on this reanalysis of earlier data in terms of currently recommended HbA(1c) targets, combination therapy with repaglinide and metformin would appear to be a good treatment option for patients with type 2 diabetes. PMID- 18405796 TI - Effects of fluticasone propionate/salmeterol combination on asthma-related health care resource utilization and costs and adherence in children and adults with asthma. AB - BACKGROUND: Clinical trials suggest that in patients with asthma inadequately controlled on low- to medium- dose inhaled corticosteroids (ICSs), the addition of a long-acting beta-agonist such as salmeterol (SAL is more effective than the addition of montelukast (MON) or a higher-dose ICS. OBJECTIVE: This study was designed to expand on these earlier findings by comparing asthma-related health care resource utilization and costs, as well as adherence to ICSs, in children and adults with asthma receiving ICS monotherapy who either were switched to fluticasone propionate plus SAL from a single inhaler (FSC) or initiated add-on therapy with SAL from a separate inhaler or MON. METHODS: This retrospective study used an integrated managed-care database from >30 health plans. Patients were >or=5 years of age with a diagnosis of asthma (International Classification of Diseases, Ninth Revision, Clinical Modification 493.xx) and >or=2 claims for FSC, SAL, or MON. The date of first claim for the medication of interest was the index date. Patients were also required to have >or=1 claim for an ICS during the 12 months preindex and 12 months postindex. Utilization and costs of asthma related care and adherence to ICS treatment postindex were compared using multivariate methods. RESULTS: After adjusting for preindex characteristics, patients receiving FSC (n=1287) had fewer claims for short-acting beta-agonists, oral corticosteroids, and lower adjusted asthma-related costs postindex compared with ICS + SAL (n=562) and ICS + MON (n=420). FSC patients also had greater adherence to ICS therapy. Those who received FSC had lower risks for treatment failure (defined as asthma-related emergency department visits or hospitalization or receipt of alternative study medication or oral corticosteroids during the postindex period). CONCLUSION: In this health insurance claims-based study of patients with asthma inadequately controlled with an ICS alone, those who received stepped-up therapy with FSC used fewer rescue medications and had greater persistence with ICSs compared with those in whom SAL or MON was added to ICS monotherapy. PMID- 18405798 TI - Words seeking action. PMID- 18405799 TI - Mentoring: the value of a personal connection. PMID- 18405800 TI - Friction and anchorage loading. PMID- 18405801 TI - Unexpected complications. PMID- 18405804 TI - Realities of mandating 3-year programs. PMID- 18405805 TI - The 3-year program issue: educational best evidence or seat of pants? PMID- 18405806 TI - Despite problems, 3-year residency has much to offer. PMID- 18405807 TI - Malocclusion and headache. PMID- 18405808 TI - Factors associated with malocclusions in children and adolescents with Down syndrome. AB - INTRODUCTION: Our aims in this study were to determine the prevalence of malocclusion stemming from vertical or transversal occlusal alterations in subjects with Down syndrome (DS) and the associations with individual, socioeconomic, and behavioral factors. METHODS: A cross-sectional study was carried out with 112 pairs of mothers and their children with DS between 3 and 18 years of age attending a genetics clinic at a public hospital in Rio de Janeiro, Brazil. Data were collected with a questionnaire given to the mothers and a clinical examination of the child or adolescent. Univariate, bivariate, and multiple logistic regression (backward stepwise) analyses were conducted. RESULTS: The results showed a prevalence of malocclusion in 74% of the sample. After the adjustment of the logistic regression model, these variables--age, nail or finger biting habit, mouth posture, and cold or sore-throat episodes in the previous 6 months--were associated with the prevalence of malocclusions in these subjects. CONCLUSIONS: The data demonstrate a high prevalence of malocclusions stemming from vertical or transversal occlusal alterations in children and adolescents with DS. Age, nail or finger biting, mouth posture, and upper airway infections were related to malocclusions in these patients. PMID- 18405809 TI - Socio-demographics of adult orthodontic visits in the United States. AB - INTRODUCTION: Population-based studies in orthodontics have focused on differences between normative and perceived needs. However, information from national data on the prevalence of orthodontic visits and their associated factors in adults in the United States is scarce. We examined the demographic profile of likely adult users of orthodontic services and whether there is racial and ethnic disparity in orthodontic visits. METHODS: We analyzed data from the Medical Expenditure Panel Survey, 2000-2004. RESULTS: Overall, about 1% of the population reported an orthodontic visit. Subjects who made a general dental visit during the current year were significantly more likely to also have an orthodontic visit. Single adults, women, people between 18 and 30 years of age, and those from high-income families were more likely to report an orthodontic visit. There were no indications of racial and ethnic disparity for either black or Hispanic adults compared with white adults after adjusting for other covariates. CONCLUSIONS: Substantial racial and ethnic disparity in adult orthodontic usage was not identified. Adults (ages 18-30 years), women, those with higher incomes, and single adults had significantly higher odds of reporting an orthodontic visit. However, additional studies specifically evaluating the association of treatment need among low-income families are required to evaluate whether these adults face significant barriers in accessing orthodontic care. PMID- 18405810 TI - Effects of surgical mandibular advancement and rotation on signs and symptoms of temporomandibular disorder: a 2-year follow-up study. AB - INTRODUCTION: The possible effects of orthognathic surgery on signs and symptoms of temporomandibular disorder (TMD) are still controversial. We prospectively investigated the association between the amount of advancement and rotation of the mandible during bilateral sagittal split osteotomy (BSSO) and the development of TMD signs and symptoms. METHODS: Class II patients (n = 127) received mandibular advancement with BSSO. We used factorial analysis of covariance to assess whether the magnitude (< or > or = 7 mm) and the direction (clockwise or counterclockwise) of the movement were associated with the onset or worsening of TMD signs and symptoms during 2 years of follow-up. RESULTS: Counterclockwise rotation of the mandible was associated with more muscle tenderness, especially in patients receiving long advancements. The combination of long advancement with counterclockwise rotation was also associated with increased joint symptoms. All symptoms declined over the 2-year follow-up period. CONCLUSIONS: Counterclockwise rotation of the mandible is related to a slight increase in muscle symptoms after BSSO. The combination of counterclockwise rotation with long advancement also might increase joint signs and symptoms. All symptomatology tended to decline over time, suggesting that the amount of advancement and mandibular rotation should not be considered as risk factors for the development of TMD in patients without preexisting conditions. PMID- 18405811 TI - Effects of Class II activator and Class II activator high-pull headgear combination on the mandible: a 3-dimensional finite element stress analysis study. AB - INTRODUCTION: The aim of this study was to evaluate the effects of the Class II activator and the Class II activator high-pull headgear (HG) combination on the mandible with 3-dimensional (3D) finite element stress analysis. METHODS: A 3D finite element model of the mandible was constructed from a dry human mandible. To investigate the effects of the Class II activator, a 3D model of the lower part of this appliance was constructed and fixed on the mandibular model. The Class II activator high-pull headgear model was established as described, and an extraoral traction force of 350 g was directed from the middle of the Class II activator to the top of the mandibular condyle. The stress regions were studied with the finite element method. RESULTS: The regions near the muscle attachment areas were affected the most. The inner part of the coronoid process and the gonial area had the maximum stress values. CONCLUSIONS: Both functional appliances can cause morphologic changes on the mandible by activating the masticatory muscles to change the growth direction. PMID- 18405812 TI - Hard- and soft-tissue contributions to the esthetics of the posed smile in growing patients seeking orthodontic treatment. AB - INTRODUCTION: The purpose of this investigation was to broaden the understanding of how various skeletal, dental, and soft-tissue relationships are related to the esthetics of the smile in patients with malocclusions before orthodontic treatment. METHODS: Images of the posed smile were captured from digital video clips of 60 growing patients (33 girls, 27 boys) seeking orthodontic treatment; they were judged by panels of laypersons and orthodontists. Discriminant analysis identified determinants of the "pleasing smile" from the results of a visual analog scale. Quantitative measurements of the soft and hard tissues were made by using the smile images, cephalometric radiographs, and study models. The esthetics of the smile were correlated with specific skeletal, dental, and soft tissue structures in the anteroposterior, vertical, and transverse dimensions (Pearson test on non-topographic correlations). RESULTS: The esthetic smile judgments of orthodontists agreed with those of laypersons (r >0.93). The vertical thicknesses of the lips were the most significant component of a pleasant smile, for both the orthodontists (upper lip) and laypersons (lower lip) (discriminant power: 75%). The vertical thickness of the upper lip had a significant positive correlation with the position of the maxillary incisor. CONCLUSIONS: Vertical lip thickness proved to be the most influential variable in smile esthetics. The significant relationship of incisor protrusion with the vertical thickness of the vermilion border of the upper lip must be considered when planning orthodontic treatment. PMID- 18405813 TI - Evaluation of facial attractiveness from end-of-treatment facial photographs. AB - INTRODUCTION: Orthodontists typically make judgments of facial attractiveness by examining groupings of profile, full-face, and smiling photographs considered together as a "triplet." The primary objective of this study was to determine the relative contributions of the 3 photographs-each considered separately-to the overall judgment a clinician forms by examining the combination of the 3. METHODS: End-of-treatment triplet orthodontic photographs of 45 randomly selected orthodontic patients were duplicated. Copies of the profile, full-face, and smiling images were generated, and the images were separated and then pooled by image type for all subjects. Ten judges ranked the 45 photographs of each image type for facial attractiveness in groups of 9 to 12, from "most attractive" to "least attractive." Each judge also ranked the triplet groupings for the same 45 subjects. The mean attractiveness rankings for each type of photograph were then correlated with the mean rankings of each other and the triplets. RESULTS: The rankings of the 3 image types correlated highly with each other and the rankings of the triplets (P <.0001). The rankings of the smiling photographs were most predictive of the rankings of the triplets (r = 0.93); those of the profile photographs were the least predictive (r = 0.76). The difference between these correlations was highly statistically significant (P = .0003). It was also possible to test the extent to which the judges' rankings were influenced by sex, original Angle classification, and extraction status of each patient. No statistically significant preferences were found for sex or Angle classification, and only 1 marginally significant preference was found for extraction pattern. CONCLUSIONS: Clinician judges demonstrated a high level of agreement in ranking the facial attractiveness of profile, full-face, and smiling photographs of a group of orthodontically treated patients whose actual differences in physical dimensions were relatively small. The judges' rankings of the smiling photographs were significantly better predictors of their rankings of the triplet of each patient than were their rankings of the profile photographs. PMID- 18405815 TI - Characterization of posed smile by using visual analog scale, smile arc, buccal corridor measures, and modified smile index. AB - INTRODUCTION: Smile evaluation and designing are important; recent emphasis on the soft tissues has been on par with that on the hard tissues in orthodontic diagnosis and treatment planning. This importance has not yet gained proper attention, and smile analysis is often ignored in clinical examinations. We attempted a comprehensive evaluation of smile characteristics with the following aims: (1) evaluation of perception differences, if any, between dental specialists and laypersons; and (2) quantification of smile characteristics with the smile arc, buccal corridor measurements, and a modified smile index (MSI). METHODS: The sixty subjects included in the study had an age range of 18 to 25 years (mean, 21 years) with equal numbers of men and women. Frontal posed smile photographs were taken of all subjects. The study had 2 parts. Analysis of perception differences between dental specialists and laypersons was performed with a visual analog scale. Quantification of smile characteristics was done with smile-arc evaluation for consonance or non-consonance, buccal corridor measurements, the MSI, and comparisons of the MSI with the facial index and the mandibular width-facial height index. RESULTS: There was no perception difference between the specialists and the laypersons on overall smile evaluation. Women had more consonant smile arcs than men, and there was high correlation between the right and left buccal corridor spaces in men and women. The MSI showed no correlation to the facial index, but there was a negative correlation of the MSI with the mandibular width-facial height index. CONCLUSIONS: Smile analysis should be an important aspect of orthodontic diagnosis and treatment planning. Orthodontists should not disturb consonant smiles but create them with proper bracket positioning. The MSI, with negative correlation with the mandibular width facial height index, warrants further evaluation with a larger sample to validate its clinical use and to develop a predictive approach of its relationship. PMID- 18405814 TI - Comparison of soft-tissue profiles after treatment with headgear or Herbst appliance. AB - INTRODUCTION: Herbst and headgear appliances are considered effective for correcting Class II malocclusions in growing patients, although their skeletal and dental effects differ. In the literature, there is no comparison between profile esthetic outcomes with the Herbst and headgear. The purpose of this study was to provide that comparison. METHODS: Lateral cephalometric radiographs of 48 matched pairs of growing Class II Division 1 patients treated with either the Herbst appliance or headgear (both combined with fixed appliances) were used to generate pretreatment and posttreatment standardized silhouettes. The silhouettes were randomly arranged and judged by lay people and orthodontic residents using a 7-point Likert scale. Statistical analyses including nonparametric procedures and intraclass correlation were used to compare initial, final, and change profile esthetic scores for the 2 groups of subjects and agreement between evaluators. RESULTS: Both groups of subjects had significant profile improvements with treatment (P <.05), and there were no statistically significant differences between the groups in average final profile scores. Overall, there was strong agreement between the evaluations of laypersons and orthodontic residents. CONCLUSIONS: Class II Division 1 growing patients treated with either Herbst appliance or headgear (both combined with fixed appliances) will benefit from significantly improved profiles that are equally attractive. PMID- 18405816 TI - Esthetic impact of premolar extraction and nonextraction treatments on Korean borderline patients. AB - INTRODUCTION: The esthetic impact of the soft-tissue profile might play a major role in deciding on premolar extraction or nonextraction treatment, particularly in borderline patients. The purpose of this study was to compare the esthetic impact of extraction and nonextraction therapy on Korean patients. METHODS: Panels of 50 dentists and 50 laypersons evaluated randomly presented pretreatment and posttreatment soft-tissue profiles of 50 extraction and 50 nonextraction patients. As subjects in this study, borderline patients were selected based on their initial diagnostic records. Two panels were asked to compare the pretreatment and posttreatment profile tracings and to quantify their perceptions of the esthetic impact of treatment change using a visual analog scale. RESULTS: Premolar extraction patients showed greater improvement in facial profile compared with nonextraction patients, regardless of the panel. Based on the results of regression analysis to determine the degree of lip protrusion at which the retraction that commonly follows premolar extraction would be needed, the intersection was about 3.0 mm behind the Ricketts' E-line; this is far behind the Korean norm. CONCLUSIONS: The results indicate that premolar extraction is more beneficial than nonextraction treatment in terms of facial profile improvement in Korean borderline patients. PMID- 18405817 TI - Esthetic evaluation of Asian-Chinese profiles from a white perspective. AB - INTRODUCTION: Immigration from Asian countries to North America and Australia has resulted in more Asian patients treated by white orthodontists. Planning treatment for patients of different ethnicities can be a challenge. The aim of this study was to establish baseline data for the assessment of Asian-Chinese profiles from a white perspective. METHODS: Two Asian-Chinese profile images, 1 male and 1 female, were modified to create 7 profiles for each gender. Each profile showed a distinct characteristic: bimaxillary protrusion, protrusive mandible, retrusive mandible, normal profile (Class I incisor with Class I skeletal pattern), retrusive maxilla, protrusive maxilla, and bimaxillary retrusion. The images were viewed and ranked by 142 white examiners in 3 cohorts: 31 orthodontists, 31 dental students, and 80 laypersons from a mixed multi-ethnic metropolitan community. The facial anatomy that played the most important role in their decision making was also noted. Collected data in the form of ranks were statistically analyzed by using repeated measures analysis of variance. RESULTS: All examiners preferred the normal Class I or bimaxillary retrusive profiles in both sexes; the male profile ranked the least attractive was the protrusive mandible, and the female profiles ranked the least attractive were the protrusive mandible and the retrusive mandible. Orthodontists, dental students, and laypersons demonstrated similar trends in ranking the profiles. The preferred profiles diverged from bimaxillary protrusive and Class III profiles (maxillary retrusion and mandibular protrusion), which are the usual norms for ethnic Asian Chinese. The upper lip, the lower lip, and the chin had the most emphasis when the examiners were ranking the profiles. This finding was also common in all 3 groups. CONCLUSIONS: This study demonstrated strong cohesive trends in establishing ideal facial esthetics of Asian-Chinese profiles evaluated by white persons in a multi-ethnic metropolitan community. PMID- 18405818 TI - Effects of extraction and nonextraction therapy with air-rotor stripping on facial esthetics in postadolescent borderline patients. AB - INTRODUCTION: A prospective randomized study was designed to compare the skeletal, dental, and soft-tissue profile changes in Class I borderline patients treated with extraction and nonextraction by means of the air-rotor stripping (ARS) technique and to compare the treatment times. METHODS: Twenty-six borderline patients with balanced and orthognathic facial profiles were randomly divided into 2 groups. In the first group, 13 subjects with a mean age of 18.1 +/ 3.7 years and mean maxillary and mandibular crowding of 5.7 +/- 1.5 and 5.9 +/- 1.4 mm, respectively, were treated by removal of 4 premolars with minimum anchorage. In the second group, 13 subjects with a mean age of 17.8 +/- 2.4 years and mean maxillary and mandibular crowding of 5.0 +/- 1.3 and 5.9 +/- 1.3 mm, respectively, were treated with ARS. Lateral cephalometric radiographs and dental models taken before and after treatment were evaluated. RESULTS: The initial skeletal, dental, and profile characteristics of both groups were similar. In the first group, the maxillary and mandibular incisors were slightly retracted, whereas lip positions did not change. In the nonextraction group, maintenance of maxillary incisor position, slight protrusion of the mandibular incisors and the upper and lower lips were observed at the end of treatment. The nasolabial angle decreased significantly. Because of postpubertal growth of the nose and the chin, the lips appeared slightly retrusive after extraction therapy, whereas lip protrusion was compensated in the nonextraction group. The main soft-tissue profile differences between the 2 groups were 1 to 1.5 mm more retruded upper and lower lip positions in the extraction patients, but both groups had well-balanced and desirable facial esthetics with all profile measurements within normal limits. Nonextraction therapy with ARS reduced treatment time by 8 months. CONCLUSIONS: Both extraction and ARS combined with nonextraction therapies are effective treatment alternatives for Class I borderline patients with good facial profile and moderate dental crowding. PMID- 18405819 TI - Long-term effectiveness of the continuous and the sectional archwire techniques in leveling the curve of Spee. AB - INTRODUCTION: The correction of a deep overbite with the subsequent achievement of long-term stability is a difficult problem for orthodontists. The role of leveling the curve of Spee (COS) in bite opening and the success of orthodontic treatment has been well documented in the literature. The aim of this study was to investigate whether leveling the COS, by using 2 orthodontic treatment techniques, produces stable results on a long-term basis. METHODS: We compared the long-term stability of leveling the COS with the straight-wire Alexander technique and the bioprogressive sectional-arch technique. The randomly selected subjects for this retrospective study were obtained from the private practices of Drs R. G. Alexander and Ruel Bench. Study models taken 2 months before treatment (T1), 2 months after treatment (T2), and after retention (T3) were evaluated. Measurements of the COS were made on the mandibular casts with a commercially available palatometer. Mandibular intercanine width, overbite, overjet, mandibular incisor irregularity, and mandibular arch length were also recorded. RESULTS AND CONCLUSIONS: Both techniques produced highly significant reductions in the COS (T1 to T2). Statistically significant, but clinically insignificant, postretention relapse of the COS occurred (T2 to T3). For both techniques, a statistically significant difference was seen in the incidence of the relapse of the COS between patients who were completely leveled posttreatment and those who were not. We did not find a correlation between pretreatment COS and relapse in any of the other occlusal traits studied. PMID- 18405820 TI - Comparison of hand-traced and computerized cephalograms: landmark identification, measurement, and superimposition accuracy. AB - INTRODUCTION: The purposes of this study were (1) to investigate the variations of landmark identification between film and digital cephalometric tracings, (2) to compare the ability of Quick Ceph 2000 (Quick Ceph Systems, Inc, San Diego, Calif) to measure the linear and angular measurements with the hand-traced method, and (3) to compare Quick Ceph 2000 superimpositions to the hand-traced method of superimpositions that are currently accepted by the American Board of Orthodontics (ABO). MATERIALS: We used 30 sets of serial cephalometric radiographs of growing patients from 1 orthodontic office. Fiduciary x- and y axes were drawn in pencil on the T1 radiographs in the regions of the cranial base, the maxilla, and the mandible. The fiduciary lines were transferred to the digital and film serial cephalograms by regionally superimposing the tracings as described in the ABO Phase III examination handbook. A Mann-Whitney test was done to compare the median and Delta of the T1 and T2 values for each measurement acquired by hand and by Quick Ceph. RESULTS AND CONCLUSIONS: There was no difference in the identification of cephalometric landmarks made manually vs digitally with Quick Ceph 2000. There was no difference in acquiring consistent cephalometric values for the measurements required by the ABO for the Phase III clinical examination manually vs digitally by using Quick Ceph 2000. There was no difference in the regional superimpositions of the mandible, the maxilla, and the cranial base, manually vs digitally with Quick Ceph 2000. PMID- 18405821 TI - Effect of a dental water jet with orthodontic tip on plaque and bleeding in adolescent patients with fixed orthodontic appliances. AB - INTRODUCTION: Effective self-care is difficult for people with orthodontic appliances because of the inherent design of brackets and archwires. It is not uncommon to have increases in plaque and gingivitis after placement of fixed appliances. The purpose of this study was to evaluate the effect of using a dental water jet (DWJ) with a specialized tip (orthodontic) on plaque and bleeding in adolescent orthodontic patients with fixed appliances. METHODS: One hundred six subjects were enrolled in this single blind, parallel clinical study. They were randomly assigned to 1 of 3 treatment groups: group 1, once daily irrigation with a DWJ and orthodontic jet tip plus a manual toothbrush; group 2, once daily flossing (FL) plus a manual toothbrush; group 3, manual toothbrush (MT) only. Plaque index (PI) and bleeding index (BI) scores were recorded at baseline, and at 2 and 4 weeks. RESULTS: All groups showed statistically significant reductions in PI (whole mouth and interproximal) at 2 and 4 weeks (P <.001). In group 1, the DWJ was statistically more effective at reducing whole mouth and interproximal plaque than the methods in the other groups (P >.001) at both 2 and 4 weeks, whereas the FL protocol in group 2 was significantly more effective than the MT protocol in group 3 at 4 weeks (P =.025) for whole-mouth plaque and at 2 and 4 weeks (P = .011 and P = .028, respectively) for interproximal plaque. All groups showed statistically significant reductions in BI (whole mouth and interproximal) at 2 and 4 weeks (P <.001). The DWJ in group 1 was statistically more effective at reducing whole-mouth bleeding than the protocols of the other groups at 2 and 4 weeks (P <.001), and the FL was statistically more effective than the MT at both times (P <.001). Both the DWJ and the FL were significantly more effective than the MT at 2 weeks (P <.001 and P <.016, respectively) for interproximal bleeding, but there were no differences between the groups at 4 weeks. CONCLUSIONS: A DWJ with a specialized orthodontic jet tip is effective for adolescents in fixed orthodontic appliances; it demonstrated beneficial results for the reduction of plaque and bleeding. PMID- 18405822 TI - Periodontal tissue activation by vibration: intermittent stimulation by resonance vibration accelerates experimental tooth movement in rats. AB - INTRODUCTION: Accelerating the speed of orthodontic tooth movement should contribute to the shortening of the treatment period. This would be beneficial because long treatment times are a negative aspect of orthodontic treatment. In this study, we evaluated the effects of mechanical stimulation by resonance vibration on tooth movement, and we showed the cellular and molecular mechanisms of periodontal ligament responses. METHODS: The maxillary first molars of 6-week old male Wistar rats were moved to the buccal side by using an expansive spring for 21 days (n = 6, control group), and the amount of tooth movement was measured. Additional vibrational stimulation (60 Hz, 1.0 m/s(2)) was applied to the first molars by using a loading vibration system for 8 minutes on days 0, 7, and 14 during orthodontic tooth movement (n = 6, experimental group). The animals were killed under anesthesia, and each maxilla was dissected. The specimens were fixed, decalcified, and embedded in paraffin. Sections were used for immunohistochemical analysis of receptor activator of NF kappa B ligand (RANKL) expression. The number of osteoclasts in the alveolar bone was counted by using TRAP staining, and the amount of root resorption was measured in sections stained with hematoxylin and eosin. RESULTS: The average resonance frequency of the maxillary first molar was 61.02 +/- 8.38 Hz. Tooth movement in the experimental group was significantly greater than in the control group (P <.05). Enhanced RANKL expression was observed at fibroblasts and osteoclasts in the periodontal ligament of the experimental group on day 3. The number of osteoclasts in the experimental group was significantly increased over the control group on day 8 (P <.05). Histologically, there were no pathological findings in either group or significant differences in the amount of root resorption between the 2 groups. CONCLUSIONS: The application of resonance vibration might accelerate orthodontic tooth movement via enhanced RANKL expression in the periodontal ligament without additional damage to periodontal tissues such as root resorption. PMID- 18405823 TI - Corrosion of orthodontic appliances--should we care? AB - Contemporary orthodontics relies on various bonded attachments, archwires, and other devices to achieve tooth movement. These components are composed of varying materials with their own distinctive physical and mechanical properties. The demands made on them are complex because they are placed under many stresses in the oral environment. These include immersion in saliva and ingested fluids, temperature fluctuations, and masticatory and appliance loading. The combination of these materials in close proximity and in hostile conditions can result in corrosion. Our purpose in this article was to consider the literature to date with regard to potential mechanical, clinical, and health implications of orthodontic corrosion. PMID- 18405824 TI - Treatment effects and anchorage potential of sliding mechanics with titanium screws compared with the Tweed-Merrifield technique. AB - The purposes of this study were to quantify the treatment effects of titanium screws on en-masse retraction of 6 anterior teeth and to compare the anchorage potential of treatment with titanium screws with the Tweed-Merrifield technique, which requires patient compliance with a high-pull J-hook. Sixteen nongrowing patients (14 women, 2 men; ages, 22.5 +/- 4.8 years) who had been treated orthodontically for bialveolar protrusion were selected. All patients had 2 maxillary first premolars extracted, 13 had mandibular first premolars extracted, and 3 had second premolars extracted. Maxillary titanium screws were placed in all patients to provide anchorage for retraction of 6 anterior teeth. Screws were placed in 8 patients to apply intrusive force to the mandibular posterior teeth. To compare the anchorage potential with a high-pull J-hook, 14 nongrowing patients (11 women, 3 men; ages, 22.9 +/- 4.0 years) who were treated with the Tweed-Merrifield technique and had an excellent compliance with a high-pull J hook were used. There was more anchorage loss of the maxillary posterior teeth in the Tweed-Merrifield group than in the titanium screw group. Both groups had excellent vertical control of the maxillary posterior teeth. There was a skeletal effect on the maxilla--reduction of A-point in the titanium screw group; this contributed to improvement of the facial profile. Treatment time in the titanium screw group was less compared with the Tweed-Merrifield technique. The success rate for the titanium screws was 87% in 25.6 +/- 5.5 months. These results suggest that titanium screws can provide acceptable and reliable anchorage and might produce skeletal effects on the maxilla. PMID- 18405825 TI - Trifocal distraction-compression osteosynthesis in conjunction with passive self ligating brackets for the reconstruction of a large bony defect and multiple missing teeth. AB - Reconstruction of a maxillary dentoalveolar defect and closure of a wide oronasal fistula in a patient with a traumatic injury are challenging for both orthodontists and surgeons. A conventional bone graft is used to fill the alveolar bone defect, to restore continuity between bony segments, and to provide bony support for tooth eruption adjacent to the defect or for orthodontic tooth movement into the bony defect. However, if the defect is too large to allow for a conventional bone graft, transport distraction osteogenesis can be used for reconstruction of the alveolar bone and implant placement. However, there is usually a discrepancy in the movement rates between the bony segment and the teeth. Passive self-ligating brackets can minimize friction between the bracket and the archwire; therefore, the rate of tooth movement can be balanced with that of the bony segment. By using orthodontic miniscrew and elastomeric traction, the regenerated bony segments can be bent to form a curved arch in the alveolar bone. In the treatment reported here, trifocal distraction-compression osteosynthesis with orthodontic miniscrews and passive self-ligating brackets helped establish bone continuity in a bony defect area, created anterior curvature of the alveolar bone, and provided good-quality regenerated bone for implant placement. PMID- 18405826 TI - Completing the 3-dimensional picture. AB - Orthodontics has a commitment to precision and accuracy similar in concept to physical engineering. Both work in a 3-dimensional (3D) world--the engineer designing and modeling physical products, and the orthodontist diagnosing and treating patients. To achieve the best possible results, both fields have readily adopted and successfully implemented highly advanced mathematical techniques and sophisticated protocols. Today, a notable difference between the 2 fields relates to the data input and analysis/modeling tools that are now available. Technology advances in the past 5 years have started to erode these barriers to safely make human 3D data input as precise and easy to obtain as physical 3D object input. Many are familiar with the advances in 3D computed tomography, specifically cone beam technology; this article focuses on a sister imaging technology called 3D surface imaging. These 3D systems enable extraoral imaging protocols to be equally precise in providing highly accurate 3D facial surface images for diagnosis, analysis, treatment monitoring, simulation, and outcome evaluation. PMID- 18405827 TI - Mini-implant anchorage for the orthodontic practitioner. AB - Mini-implant-enhanced anchorage has become a popular concept in orthodontics over the past years. Although these systems are routinely used in university settings, there is some reservation because of lack of information in private practices. This article will introduce the concept of mini-implant anchorage to the orthodontic practitioner. PMID- 18405828 TI - An insight into the sialotranscriptome of the non-blood feeding Toxorhynchites amboinensis mosquito. AB - All adult mosquitoes take sugar meals, and most adult females also take blood meals to develop eggs. Salivary glands (SG) of males are thus much smaller and do not contain many of the antihemostatic and antiinflammatory compounds found in females. In the past 5 years, transcriptome analyses have identified nearly 70 different genes expressed in adult female SG. For most of these, no function can be assigned in either blood or sugar feeding. Exceptionally, Toxorhynchites mosquitoes are unusual in that they never feed on blood, and the SG of adults are identical in both sexes. Transcriptome analysis of the adult SG of this mosquito was performed to increase knowledge of the evolution of blood feeding--and to identify polypeptide families associated with sugar feeding--in mosquitoes. PMID- 18405829 TI - Developmental expression patterns of cuticular protein genes with the R&R Consensus from Anopheles gambiae. AB - CPR proteins are the largest cuticular protein family in arthropods. The whole genome sequence of Anopheles gambiae revealed 156 genes that code for proteins with the R&R Consensus and named CPRs. This protein family can be divided into RR 1 and RR-2 subgroups, postulated to contribute to different regions of the cuticle. We determined the temporal expression patterns of these genes throughout post-embryonic development by means of real-time qRT-PCR. Based on expression profiles, these genes were grouped into 21 clusters. Most of the genes were expressed with sharp peaks at single or multiple periods associated with molting. Genes coding for RR-1 and RR-2 proteins were found together in several co expression clusters. Twenty-five genes were expressed exclusively at one metamorphic stage. Five out of six X-linked genes showed equal expression in males and females, supporting the presence of a gene dosage compensation system in A. gambiae. Many RR-2 genes are organized into sequence clusters whose members are extremely similar to each other and generally closely associated on a chromosome. Most genes in each sequence cluster are expressed with the same temporal expression pattern and at the same level, suggesting a shared mechanism to regulate their expression. PMID- 18405830 TI - Mutations in Dalpha1 or Dbeta2 nicotinic acetylcholine receptor subunits can confer resistance to neonicotinoids in Drosophila melanogaster. AB - Resistance to insecticides by modification of their molecular targets is a serious problem in chemical control of many arthropod pests. Neonicotinoids target the nicotinic acetylcholine receptor (nAChR) of arthropods. The spectrum of possible resistance-conferring mutations of this receptor is poorly understood. Prediction of resistance is complicated by the existence of multiple genes encoding the different subunits of this essential component of neurotransmission. We focused on the cluster of three Drosophila melanogaster nAChR subunit genes at cytological region 96A. EMS mutagenesis and selection for resistance to nitenpyram was performed on hybrids carrying a deficiency for this chromosomal region. Two complementation groups were defined for the four strains isolated. Molecular characterisation of the mutations found lesions in two nAChR subunit genes, Dalpha1 (encoding an alpha-type subunit) and Dbeta2 (beta-type). Mutations conferring resistance in beta-type receptors have not previously been reported, but we found several lesions in the Dbeta2 sequence, including locations distant from the predicted neonicotinoid-binding site. This study illustrates that mutations in a single-receptor subunit can confer nitenpyram resistance. Moreover, some of the mutations may protect the insect against nitenpyram by interfering with subunit assembly or channel activation, rather than affecting binding affinities of neonicotinoids to the channel. PMID- 18405831 TI - The 20-hydroxyecdysone-induced signalling pathway in G2/M arrest of Plodia interpunctella imaginal wing cells. AB - The mechanisms involved in the control of cellular proliferation by the steroid hormone 20-hydroxyecdysone (20E) in insects are not known. We dissected the 20E signalling pathway responsible for G2/M arrest of imaginal cells from the IAL PID2 cells of the Indian meal moth Plodia interpunctella. We first used a 5'-3' RACE-based strategy to clone a 4479bp cDNA encoding a putative P. interpunctella HR3 transcription factor named PiHR3. The deduced amino acid sequence of PiHR3 was highly similar to those of HR3 proteins from other lepidopterans, e.g. Manduca sexta and Bombyx mori. Using double-stranded RNA-mediated interference (dsRNAi), we then succeeded in blocking the ability of 20E to induce the expression of PiEcR-B1, PiUSP-2 and PiHR3 genes that encode the P. interpunctella ecdysone receptor B1-isoform, Ultraspiracle-2 isoform, the insect homologue of the vertebrate retinoid X receptor, and the HR3 transcription factor. We showed that inhibiting the 20E induction of PiEcR-B1, PiUSP-2 and PiHR3 mRNAs prevented the decreased expression of B cyclin and consequently the G2/M arrest of IAL-PID2 cells. Using this functional approach, we revealed the participation of EcR, USP and HR3 in a 20E signalling pathway that controls the proliferation of imaginal cells by regulating the expression of B cyclin. PMID- 18405832 TI - Cross-induction of detoxification genes by environmental xenobiotics and insecticides in the mosquito Aedes aegypti: impact on larval tolerance to chemical insecticides. AB - The effect of exposure of Aedes aegypti larvae to sub-lethal doses of the pyrethroid insecticide permethrin, the organophosphate temephos, the herbicide atrazine, the polycyclic aromatic hydrocarbon fluoranthene and the heavy metal copper on their subsequent tolerance to insecticides, detoxification enzyme activities and expression of detoxification genes was investigated. Bioassays revealed a moderate increase in larval tolerance to permethrin following exposure to fluoranthene and copper while larval tolerance to temephos increased moderately after exposure to atrazine, copper and permethrin. Cytochrome P450 monooxygenases activities were induced in larvae exposed to permethrin, fluoranthene and copper while glutathione S-transferase activities were induced after exposure to fluoranthene and repressed after exposure to copper. Microarray screening of the expression patterns of all detoxification genes following exposure to each xenobiotic with the Aedes Detox Chip identified multiple genes induced by xenobiotics and insecticides. Further expression studies using real time quantitative PCR confirmed the induction of multiple CYP genes and one carboxylesterase gene by insecticides and xenobiotics. Overall, this study reveals the potential of xenobiotics found in polluted mosquito breeding sites to affect their tolerance to insecticides, possibly through the cross-induction of particular detoxification genes. Molecular mechanisms involved and impact on mosquito control strategies are discussed. PMID- 18405833 TI - Pheromone biosynthetic pathways: PBAN-regulated rate-limiting steps and differential expression of desaturase genes in moth species. AB - We combine the use of labeled precursors with enzyme inhibitors to decipher the biosynthetic pathway of pheromone biosynthesis and the rate-limiting step/s that are regulated by pheromone biosynthesis activating neuropeptide (PBAN). We demonstrate that Plodia interpunctella is able to utilize hexadecanoic acid, and to a lesser extent tetradecanoic acid, for the biosynthesis of the main pheromone component (Z,E)-9,12-tetradecadienyl acetate. This indicated that the main pathway involves a Delta11 desaturase, chain shortening, followed by a Delta12 desaturase, but that a functional Delta9 desaturase could also be utilized. Using reverse transcription-quantitative real-time polymerase chain reaction (RT-QPCR) we distinguish two out of nine possible desaturase gene transcripts in P. interpunctella that are expressed at the highest levels. The rate-limiting step for PBAN-stimulation was studied in two moth species so as to compare the biosynthesis of a diene (P. interpunctella) and a monoene (Helicoverpa armigera) main pheromone component. In both species, incorporation of label from the (13)C sodium acetate precursor was activated by PBAN whereas no stimulatory action was observed in the incorporation of the precursors: (13)C malonyl coenzyme A; hexadecanoic 16,16,16-(2)H(3) or tetradecanoic 14,14,14-(2)H(3) acids. The acetyl coenzyme A carboxylase (ACCase) inhibitor, Tralkoxydim, inhibited the PBAN stimulation of incorporation of stable isotope whereas the fatty-acyl reductase inhibitor, Mevastatin, failed to influence the stimulatory action of PBAN. These results provide irrefutable support to the hypothesis that PBAN affects the production of malonyl coenzyme A from acetate by the action of ACCase in the pheromone glands of these moths. PMID- 18405834 TI - Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus) microplus. AB - The cellular and molecular characteristics of a cell line (BME26) derived from embryos of the cattle tick Rhipicephalus (Boophilus) microplus were studied. The cells contained glycogen inclusions, numerous mitochondria, and vesicles with heterogeneous electron densities dispersed throughout the cytoplasm. Vesicles contained lipids and sequestered palladium meso-porphyrin (Pd-mP) and rhodamine hemoglobin, suggesting their involvement in the autophagic and endocytic pathways. The cells phagocytosed yeast and expressed genes encoding the antimicrobial peptides (microplusin and defensin). A cDNA library was made and 898 unique mRNA sequences were obtained. Among them, 556 sequences were not significantly similar to any sequence found in public databases. Annotation using Gene Ontology revealed transcripts related to several different functional classes. We identified transcripts involved in immune response such as ferritin, serine proteases, protease inhibitors, antimicrobial peptides, heat shock protein, glutathione S-transferase, peroxidase, and NADPH oxidase. BME26 cells transfected with a plasmid carrying a red fluorescent protein reporter gene (DsRed2) transiently expressed DsRed2 for up to 5 weeks. We conclude that BME26 can be used to experimentally analyze diverse biological processes that occur in R. (B.) microplus such as the innate immune response to tick-borne pathogens. PMID- 18405835 TI - An abundant acyl-CoA (Delta9) desaturase transcript in pheromone glands of the cabbage moth, Mamestra brassicae, encodes a catalytically inactive protein. AB - The principal sex pheromone component produced by females of the cabbage moth, Mamestra brassicae, is derived from the monounsaturated fatty acid, Z11-16:1, whereas two additional trace components are derived from E11-16:1 and Z9-16:1. This report presents the isolation and analysis of cDNAs encoding pheromone gland specific acyl-CoA desaturases implicated in the production of these unsaturated fatty acids (UFAs). Comparisons of the encoded amino acid sequences of four cDNA fragments isolated by degenerate PCR from cabbage moth pheromone glands established their orthology with previously characterized noctuid desaturases as follows: MbraLPAQ, belonging to the pheromone gland-specific LPAQ desaturase lineage having Delta11 regioselectivity, MbraKPSE-a and MbraKPSE-b, belonging to the pheromone gland-specific KPSE desaturase lineage having Delta9 regioselectivity and a substrate preference for palmitic acid (16:0) over oleic acid (18:0), and MbraNPVE, belonging to the NPVE desaturase lineage having Delta9 regioselectivity and a substrate preference 18:0>16:0. Full-length cDNAs corresponding to the two most abundantly expressed pheromone gland-specific desaturase transcripts, MbraLPAQ and MbraKPSE-b, were isolated and assayed for their ability to genetically complement the UFA auxotrophy of a desaturase deficient ole1 strain of Saccharomyces cerevisiae. The MbraLPAQ desaturase restored UFA prototrophy and GC-MS analysis identified Z11-16:1 and Z11-18:1 as the predominant UFAs produced. Surprisingly, MbraKPSE-b failed to complement the ole1 mutation, although it shares >98% amino acid sequence similarity with other noctuid KPSE desaturases that do. Site-directed mutagenesis of either or both of two nonconservative amino acid substitutions restored functionality to the MbraKPSE-b protein, although GC-MS analysis revealed that neither reversion resulted in the characteristic KPSE substrate preference for 16:0. PMID- 18405836 TI - Molecular and functional characterization of granulin-like molecules of insects. AB - Granulins are a group of highly conserved growth factors that have been described from a variety of organisms spanning the metazoa. Here, we report on the identification of two partial transcripts encoding granulin-like molecules from Aa23 embryonic cells of Aedes albopictus and primary haemocytes from Manduca sexta. Both these partial transcripts had the characteristic 12-cysteine motif that is a hallmark of the granulin family and they represent the first granulin mRNA transcripts identified from insects. Moreover, we demonstrate that the recombinant granulin molecule that we originally cloned in the goldfish, induced proliferation of both Aa23 embryonic cells and primary haemocytes. Interestingly, this proliferative effect was upregulated in the presence of the intracellular symbiotic bacterium Wolbachia pipientis. Thus, granulin appears to be a highly conserved growth factor not only in lower vertebrates but also invertebrates. PMID- 18405837 TI - Molecular and functional characterization of voltage-gated sodium channel variants from Drosophila melanogaster. AB - Extensive alternative splicing and RNA editing have been documented for the transcript of DmNa(V) (formerly para), the sole sodium channel gene in Drosophila melanogaster. However, the functional consequences of these post-transcriptional modifications are not well understood. In this study we isolated 64 full-length DmNa(V) cDNA clones from D. melanogaster adults. Based on the usage of 11 alternative exons, 64 clones could be grouped into 29 splice types. When expressed in Xenopus oocytes, 33 DmNa(V) variants generated sodium currents large enough for functional characterization. Among these variants, DmNa(V)5-1 and DmNa(V)7-1 channels activated at the most hyperpolarizing potentials, whereas DmNa(V)1-6 and DmNa(V)19 channels activated at the most depolarizing membrane potentials. We identified an A-to-I editing event in DmNa(V)5-1 that is responsible for its uniquely low-voltage-dependent activation. The wide range of voltage dependence of gating properties exhibited by DmNa(V) variants represents a rich resource for future studies to determine the role of DmNa(V) in regulating sodium channel gating, pharmacology, and neuronal excitability in insects. PMID- 18405838 TI - Whither antibacterial drug discovery? PMID- 18405840 TI - Assessing drug-likeness--what are we missing? AB - The concept of drug-likeness helps to optimise pharmacokinetic and pharmaceutical properties, for example, solubility, chemical stability, bioavailability and distribution profile. A number of molecular descriptors have emerged as reasonably informative and predictive, for example, the Rule-of-Five. Here, we review some current approaches, then discuss their major shortcoming, namely the static nature of the structural features and physicochemical properties they encode. As we demonstrate, molecules are not 'frozen statues' but 'dancing ballerinas', and several of their computable physicochemical properties are conformation-dependent and lead to the concept of property spaces. Molecular sensitivity (namely, how much a given computable physicochemical property varies as a function of flexibility) appears as a promising descriptor to encode some of the information contained in molecular property spaces. PMID- 18405839 TI - The conductor and his orchestra. PMID- 18405841 TI - Kinases as drug targets in the treatment of bipolar disorder. AB - Bipolar disorder is one of the most severely debilitating of all medical illnesses, and is increasingly recognized as a major public health problem. For many patients with bipolar disorder, current pharmacotherapy is insufficient. Exciting recent data suggest that regulation of signaling molecules may be involved in the pathophysiology of the disorder, and in the mechanisms of action of mood stabilizers and antidepressants. Through our developing understanding of the biochemical targets of effective medications, several potential targets for new therapies have emerged. This short review will focus on two of the most promising such targets: glycogen synthase-3 and protein kinase C. PMID- 18405842 TI - Computational toxicology in drug development. AB - Computational tools for predicting toxicity have been envisaged for their potential to considerably impact the attrition rate of compounds in drug discovery and development. In silico techniques like knowledge-based expert systems (quantitative) structure activity relationship tools and modeling approaches may therefore help to significantly reduce drug development costs by succeeding in predicting adverse drug reactions in preclinical studies. It has been shown that commercial as well as proprietary systems can be successfully applied in the pharmaceutical industry. As the prediction has been exhaustively optimized for early safety-relevant endpoints like genotoxicity, future activities will now be directed to prevent the occurrence of undesired toxicity in patients by making these tools more relevant to human disease. PMID- 18405843 TI - Computational models for prediction of interactions with ABC-transporters. AB - The polyspecific ligand recognition pattern of ATB-binding cassette (ABC) transporters, combined with the limited knowledge on the molecular basis of their multispecificity, makes it difficult to apply traditional molecular modelling and quantitative structure-activity relationships (QSAR) methods for identification of new ligands. Recent advances relied mainly on pharmacophore modelling and machine learning methods. Structure-based design studies suffer from the lack of available protein structures at atomic resolution. The recently published protein homology models of P-glycoprotein structure, based on the high-resolution structure of the bacterial ABC-transporter of Sav1866, may open a new chapter for structure-based studies. Last, but not least, molecular dynamics simulations have already proved their high potential for structure-function modelling of ABC transporter. Because of the recognition of several ABC-transporters as antitargets, algorithms for predicting substrate properties are of increasing interest. PMID- 18405844 TI - Screening isolates from antibody phage-display libraries. AB - Antibody phage display, coupled with automated screening, facilitates and potentiates the mining of complex combinatorial libraries and the identification of potent drug leads. In managing phage screening data, the behavior of individual phage isolates in binding assays must be linked to their antibody identities as deduced from DNA sequencing. Reviewed here are recently reported approaches for high-throughput screening of clones isolated from phage antibody libraries after selection on a defined antigen. Specific information management challenges, and possible solutions, are described for organizing screening data to enable rapid lead discovery using these antibody libraries. PMID- 18405845 TI - Serotoninergic mechanisms in the treatment of obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) is a disabling psychiatric condition affecting 1-2% of the community. Although modern drug, behavioral and psychosurgical therapies have improved the prognosis of OCD considerably, approximately 30% of patients remain treatment-refractory. Currently, selective serotonin (5-HT) reuptake inhibitors (SSRIs) are the drug treatments of choice for OCD. Accordingly, this review evaluates the evidence for a role of the serotonin (5-HT) neurochemical system in the treatment and pathophysiology of OCD. However, drug treatment approaches that modify function of interrelated neurochemical systems, such as the dopamine and glutamate systems, are also briefly discussed as they promise to complement and enhance SSRI treatment effects. PMID- 18405846 TI - Pharmacological treatments of fibromyalgia: do complex conditions need complex therapies? AB - Fibromyalgia (FM) is a chronic pain condition, with auxiliary symptoms, such as sleep disturbances and fatigue. Although many of the mechanisms of action targeted by the drugs used to treat FM have been focused to the management of single symptoms, drugs (e.g. pregabalin, duloxetine) have now been identified that demonstrate a multidimensional effect. However, such drugs often fail to demonstrate acceptable efficacy in the majority of the patient population. Thus, the mechanisms of action of the drugs studied as treatments for FM are either identifying subgroups within the pathophysiology of the condition or suggesting that a mechanism of action that will offer universal efficacy has, as yet, to be identified. PMID- 18405847 TI - PK/PD modelling and simulations: utility in drug development. AB - Pharmacokinetic/pharmacodynamic (PK/PD) modelling and simulation can be used as an 'applied science' tool to provide answers on efficacy and safety of new drugs faster and at a lower cost. PK/PD modelling can be used from the preclinical phase through all clinical phases of drug development. Optimal use of PK/PD modelling and simulation will lead to fewer failed compounds, fewer study failures and smaller numbers of studies needed for registration. For PK/PD modelling to fulfil its potential in drug development, it needs to be embraced across the industry and regulatory agencies, and more education on this topic is required. PMID- 18405848 TI - Impact of non-profit organizations on drug discovery: opportunities, gaps, solutions. AB - Non-profit organizations (NPO) play an increasingly important role in drug discovery and development for diseases that are neglected by the pharmaceutical industry because of low or absent commercial incentives. Governments and major private foundations such as the Wellcome Trust and the Bill & Melinda Gates Foundation increasingly step in to provide strategic direction, communication platforms and major resources, motivated by the fact that major healthcare problems remain unsolved. Drug discovery in the field of neglected diseases is fraught with complexities since, in many cases, important tools are lacking including readily available diagnostics, molecular epidemiology, appropriate model systems, representative strain collections, biomarkers, up-to-date trial methodologies and regulatory strategies. On top of this, the high hurdles addressing novel drug targets must be cleared. PMID- 18405849 TI - The role of users in innovation in the pharmaceutical industry. AB - Traditionally, innovation in the pharmaceutical industry is organised according to the linear model. Over the past two decades this model lost its meaning as a result of rising costs, increased competition, new scientific developments and better-informed, more demanding users. The linear model is not well equipped to involve these new actors and to include their feedback. Starting from a systemic approach, the involvement of actors in pharmaceutical innovation processes, more in particular users, is put central. It is discussed and illustrated with three cases why a systemic model may be more effective to cope with present developments and why users should be involved. To wind up, conclusions are drawn regarding the implications of a systemic approach for policymakers, researchers and firms. PMID- 18405850 TI - Snapshot PK: a rapid rodent in vivo preclinical screening approach. AB - Described in this article are strategies implemented to increase the throughput of in vivo rodent pharmacokinetic (PK) studies using the snapshot PK study design and automated methods for compound submission, sample processing, data analysis and reporting. Applying snapshot PK studies to categorize the oral exposure of >1300 discovery compounds as low, moderate or high resulted in an attrition rate of 86%. The follow up full PK studies on the remaining compounds found that 98% of the compounds were predicted in the correct (69%) or adjacent (29%) oral exposure category by the snapshot PK studies. These results demonstrate that the snapshot PK screen in rodents can serve as an effective and efficient in vivo tool in the compound selection process in drug discovery. PMID- 18405852 TI - Rituximab for follicular non-Hodgkin lymphoma. PMID- 18405853 TI - Carcinogenicity of some aromatic amines, organic dyes, and related exposures. PMID- 18405860 TI - Part I: Cancer in continuing dramas. PMID- 18405861 TI - A nexus of science and clinical immunology: the Federation of Clinical Immunology Societies and the FOCIS Centers of Excellence. PMID- 18405862 TI - Tolerance: it's in your blood. PMID- 18405863 TI - Focus on FOCIS: interleukin 2 treatment associated autoimmunity. AB - A patient from the University of Pittsburgh is presented who developed autoimmunity during IL-2 based combination therapy. IL-2 was originally described as a "T cell growth factor" capable of expanding previously activated T cells, enhancing the cytotoxicity of antigen-specific cytotoxic T cells and NK cells. High dose Interleukin 2 (HDIL2) is now FDA-approved for therapy of patients with metastatic melanoma and renal cell carcinoma, based on its ability to induce durable responses in 5-10% of patients. The antitumor effect of HDIL2 is incompletely understood, but it appears that this regimen alters the balance of immigrant T effector cells in relation to T suppressor cells. It promotes a less immunosuppressive tumor microenvironment, inducing tumor regression in a subset of patients that is yet to be defined. The antitumor activity of IL-2, as for other agents that promote durable antitumor activity against melanoma such as interferon alpha and anti-CTLA4 antibody, is frequently associated with development of autoimmunity as observed in this patient. We present studies investigating the mechanisms for the therapeutic benefit of HDIL2 in melanoma. PMID- 18405864 TI - Investment in body image for self-definition results in greater vulnerability to the thin media than does investment in appearance management. AB - This study investigated the effect of thin images according to two dimensions of body-image (BI) investment. Ninety-five females were classified as high or low investors based on the Appearance Schemas Inventory-Revised Self-Evaluative Salience (SES) and Motivational Salience (MS) subscales. Participants viewed advertisements portraying either the thin ideal or products. Results indicated that both women high in SES and MS reported lowered appearance self-esteem but greater BI importance following thin exposures. However, only the high SES group reported greater BI dissatisfaction and importance of current-ideal discrepancies after seeing thin images. Although highly invested women (regardless of their motivation for investment) are more responsive to thin media images than are women low in investment, those invested for self-definition are affected on more dimensions than are those invested for appearance management. PMID- 18405865 TI - Susceptibility for thin ideal media and eating styles. AB - This study examined the relations between susceptibility for thin ideal media and restrained, emotional and external eating, directly and indirectly through body dissatisfaction. Thin ideal media susceptibility, body dissatisfaction and eating styles were measured in a sample of 163 female students. Structural equation modelling was used for analyses, controlling for BMI. Higher susceptibility for thin ideal media was directly related to higher scores on all eating styles, and indirectly related to higher restrained and emotional eating through elevated levels of body dissatisfaction. So, thin ideal media susceptibility was not only related to restraint through body dissatisfaction, but also directly. Emotional eaters might be more vulnerable for negative affect, whereas external eaters might be more sensitive to external cues in general. PMID- 18405866 TI - Implications of accuracy, sensitivity, and variability of body size estimations to disordered eating. AB - The current study was conducted to investigate the relationships between body size estimations and disordered eating symptomatology. The method of constant stimuli was used to derive three measures of self-perceived body size in 93 women: (1) accuracy of body size estimations (body image distortion); (2) sensitivity in discriminating body size within blocks of trials (body image sensitivity); and (3) variability in making body size estimations between blocks of trials (body image variability). Participants also completed measures of disordered eating. Although body image distortion correlated with dietary restraint and eating concern, body image variability accounted for additional variance in these variables, as well as variance in binge eating. The relationships involving body image variability were found to be mediated by body dissatisfaction and internalization of the thin ideal. Together, these results are consistent with the proposition that body image variability is a significant factor in disordered eating. PMID- 18405867 TI - Body checking and avoidance among behavioral weight-loss participants. AB - The present study examined the psychological and weight-related correlates of body checking and avoidance behaviors in individuals who have lost weight through obesity treatment. Among 185 individuals in behavioral weight-control treatment, the prevalence of body checking and avoidance behavior was assessed, as well as the relationship of checking and avoidance to attitudinal disturbances, weight loss, and perception of struggling in treatment. Checking and avoidance were frequent, and they correlated significantly with higher overvaluation of shape and weight, lower self-esteem, greater body dissatisfaction, higher fear of fat, and the perception of greater struggling in weight-loss treatment. Checking was inversely correlated with the percentage weight lost in treatment and BMI lost but not related to pre-treatment or current BMI. Thus, body checking and avoidance behaviors were associated with psychological disturbances and poorer treatment outcome. PMID- 18405868 TI - Positive illusions about one's partner's physical attractiveness. AB - This study examined couples' ratings of self and partner physical attractiveness. On the basis of the theory of positive illusions, it was expected that individuals would rate their partners as more attractive than their partners would rate themselves. Both members of 93 heterosexual couples, with a mean relationship length of about 14 years, provided ratings of both their own and their partner's physical attractiveness. Results support the theory that individuals hold positive illusions about their partner's physical attractiveness. Implications of these results in terms of relationship-enhancing biases are discussed. PMID- 18405869 TI - Psychological and behavioral correlates of the SATAQ-3 with males. AB - The current study used confirmatory factor analysis (CFA) to investigate the factor structure of a revised version of the Sociocultural Attitudes Towards Appearance Questionnaire (SATAQ-3) with a sample of undergraduate males. In addition to completing the SATAQ-3, participants completed various measures pertaining to body image, behavioral, and psychological constructs to examine concurrent and discriminant validity. CFA results indicated that the four factors of the SATAQ-3, Information, Pressures, Internalization-General, and Internalization-Athlete, represented the data well and better than an alternative 3-factor structure that combined the two Internalization subscales. All four factors demonstrated excellent concurrent and discriminant validity. Findings from this study support use of a slightly modified version of the SATAQ-3 with males. The present results also support the continued investigation of media influences as multidimensional constructs important to male body image. PMID- 18405870 TI - The Sociocultural Attitudes Towards Appearance Questionnaire-3: a confirmatory factor analysis. AB - The Sociocultural Attitudes Towards Appearance Questionnaire-3 measures awareness and endorsement of societal appearance standards. The instrument has been subjected to exploratory factor analyses but to date no studies have reported a priori tests of its hypothesized factor structure using confirmatory factor analysis (CFA). The aim of the present study was to subject the SATAQ-3 to a CFA. Results from a non-clinical convenience sample of 369 women revealed an adequate fit of the model according to conventional criteria. However, detailed residual analysis indicated a significant lack of fit which was explainable by one mis specified item and shared method variance due to similarities in item content. It was concluded that, with the removal of the mis-specified item, the degree of misfit was tolerable and the intended four-factor solution provides a satisfactory and parsimonious representation of the data. PMID- 18405871 TI - German translation and psychometric evaluation of the Body Appreciation Scale. AB - This paper examined the psychometric properties of the German version of the Body Appreciation Scale (BAS), a novel scale for the assessment of positive body image. Based on a community sample of 156 women and 144 men in Austria, results showed good internal reliability and construct validity for the BAS scores, as well as a unidimensional factor structure for both women and men. Specifically, Cronbach's alpha was high for both women (alpha=.90) and men (alpha=.85), and the BAS was correlated with the body esteem scale and self-esteem. In addition, women with lower BMIs reported greater body appreciation, but no such association was found for men. Finally, there were small sex differences in BAS scores, with men scoring more positively than women (Cohen's d=0.26). The German BAS, a useful indicator of positive body image among Austrian adults, should prove valuable for the assessment of body image in German-speaking countries. PMID- 18405872 TI - Post-hepatectomy regeneration of the murine liver I. Effect upon Schistosoma mansoni lesions, before and after chemotherapy. AB - Hepatic histopathological changes due to Schistosoma mansoni infection in the mouse presented considerable improvement following partial hepatectomy, both during early (acute) and late (chronic) infections, and especially when surgery was preceded by curative chemotherapy. A 60% hepatectomy removed a great deal of a diseased liver that was replaced by a normal-looking tissue in which schistosomal lesions appeared fewer and scattered. After chemotherapy, residual fibrosis left either from cured acute and chronic schistosomal lesions, almost completely disappeared when the regenerated liver was examined a month afterwards. These marked changes, brought about by hepatectomy in experimental hepatic schistosomiasis, illustrate the fact that post-hepatectomy regeneration tends to restore the normal structure of the liver, even in a diseased organ. PMID- 18405873 TI - Truncation of the Down syndrome candidate gene DYRK1A in two unrelated patients with microcephaly. AB - We have identified and characterized two unrelated patients with prenatal onset of microcephaly, intrauterine growth retardation, feeding problems, developmental delay, and febrile seizures/epilepsy who both carry a de novo balanced translocation that truncates the DYRK1A gene at chromosome 21q22.2. DYRK1A belongs to the dual-specificity tyrosine phosphorylation-regulated kinase (DYRK) family, which is highly conserved throughout evolution. Given its localization in both the Down syndrome critical region and in the minimal region for partial monosomy 21, the gene has been studied intensively in animals and in humans, and DYRK1A has been proposed to be involved in the neurodevelopmental alterations associated with these syndromes. In the present study, we show that truncating mutations of DYRK1A result in a clinical phenotype including microcephaly. PMID- 18405874 TI - Prevention of anterior capsule contraction by anterior capsule relaxing incisions with neodymium:yttrium-aluminum-garnet laser. AB - PURPOSE: To examine the effect of two or three neodymium:yttrium-aluminum-garnet (Nd:YAG) laser relaxing incisions made in the anterior capsular rim on prevention of anterior capsule contraction after cataract surgery. DESIGN: Randomized clinical trials. METHODS: One hundred patients scheduled for bilateral cataract surgery were randomized to one of two groups: two or three relaxing incisions in the left eye and no incisions in the right eye, and relaxing incisions in the right eye and no incisions in the left eye. The anterior capsule opening area was measured using Scheimpflug photography immediately after capsulotomy and at one, three, and six months after capsulotomy, and the percentage reduction was calculated. The degree of intraocular lens (IOL) decentration and tilt and of posterior capsule opacification (PCO) also were examined. RESULTS: There was no significant difference in the anterior capsule opening area between fellow eyes at baseline (P > or = .1770). In patients who underwent two incisions, no significant difference was found between fellow eyes in the opening area (P > or = .4098) or in the percentage of reduction (P > or = .8730) throughout follow-up. In patients who underwent three incisions, the opening area in eyes with capsulotomy was significantly greater than that in eyes without capsulotomy (P < or = .0154), and the percentage reduction was significantly smaller in eyes with capsulotomy than in eyes without capsulotomy (P < or = .0016). No significant differences were found in the IOL decentration and tilt, or in PCO. CONCLUSIONS: Three relaxing incisions made in the anterior capsule decrease the anterior capsule contraction, whereas two incisions do not. PMID- 18405875 TI - Ocular adnexal lymphoid tumors: progress in need of clarification. AB - PURPOSE: To evaluate recent ophthalmic publications on ocular adnexal lymphomas (OALs) according to histopathologic and immunophenotypic criteria used in the diagnosis of systemic lymphomas (World Health Organization classification). DESIGN: Summary and critical analysis of recent clinical and pathologic studies. METHODS: Literature review and interpretation. RESULTS: In the largest study of 353 cases of OALs published in the pathology literature, extranodal marginal zone lymphoma (EMZL) constituted 52%, follicular lymphoma 23%, mantle cell lymphoma 5%, and small cell lymphocytic and chronic lymphocytic leukemia 4%-plus a residuum of arcane entities. In smaller series of less intensively studied OALs in the ophthalmic literature, EMZLs had a higher preponderance and also were associated with a favorable prognosis. Many EMZLs seemed to arise primarily in the ocular adnexa (mucosa-associated lymphoid tissue [MALT] lymphoma should be restricted for EMZLs involving epithelial tissues). CONCLUSIONS: Rigorous diagnostic criteria and a proposal for a prospective multicenter study may bring further clarification to the emerging order in this set of tumors. PMID- 18405877 TI - [Management of thyroid dysfunction by primary care physicians of the Province of Quebec: a transversal study]. AB - OBJECTIVES: This study aims to evaluate clinical practice of primary care physicians regarding common thyroid disorders. MATERIALS AND METHODS: A sample of 210 primary care physicians was randomly selected in three Quebec's administrative regions. Four clinical vignettes (V1 to V4) were presented by mail: two cases of subclinical hypothyroidism (women of 25 years - V1 - and 70 - V2 - years of age) for which physicians had to choose to either treat or not with thyroid replacement and two cases of hyperthyroidism (women of 30 - V3 - and 66 - V4- years of age) for which they had to choose a course of action (observation, treatment or referral to a specialist). V1 and V2 where followed by four sub questions presenting supportive elements that could influence the decision to treat (presence of antithyroid antibodies, accumulation of symptoms, LDL cholesterol and thyreostimulin levels). RESULTS: The overall response rate was 22%. Forty-two percent of respondents would have treated V1 outright and 49% would have treated V2. The therapeutic approach in the face of these two vignettes, independently of the presence or absence of supportive clinical or biochemical elements, did not vary according to geographic practice area. However, one region was significantly more conservative for V4. The number of years in practice or assistance to continuous medical education activities did not affect management of vignettes. CONCLUSION: This study outlines the importance of clinical practice guidelines and tools to facilitate their application in clinical management of thyroid disorders. PMID- 18405876 TI - Retinal sensitivity measured with the micro perimeter 1 after resolution of central serous chorioretinopathy. AB - PURPOSE: To examine retinal sensitivity in the macular area of eyes with resolved central serous chorioretinopathy (CSC). DESIGN: Retrospective chart review. METHODS: We studied retrospectively the medical records of 21 patients (21 eyes) with resolved CSC. Using spectral-domain optical coherence tomography (SD-OCT), 6 x 6-mm areas of macula were examined with 256 sequential horizontal scans. Microperimetry in the macular area was performed with the Micro Perimeter 1 (Nidek, Vigonza, Italy). Seventy-two measurement points were located within the central 10 degrees of the macula. RESULTS: In eyes with resolved CSC, most OCT images showed a physiologically normal appearance. However, irregularities of the retinal pigment epithelium (RPE) were detected in 11 eyes, and focal defects of the junctions between inner and outer segments (IS/OS) of the photoreceptors were noted in 15 eyes. These abnormalities often were seen within the foveal region. At 983 (79.8%) of 1,232 points with intact retina, retinal sensitivity was 16 decibels (dB) or better. However, a retinal sensitivity of 16 dB or better was obtained at 34.0% of points with irregular RPE and in 20.0% of points with defects of the IS/OS. The mean retinal sensitivities within the area with irregular RPE (13.4 +/- 4.8 dB) or with defects of the IS/OS (11.5 +/- 4.2 dB) were significantly lower than that of intact retina (17.5 +/- 2.4 dB; P < .0001). CONCLUSIONS: Eyes with resolved CSC often show focal areas with reduced retinal function, which are consistent with irregularity of the RPE or with defects of the IS/OS. PMID- 18405878 TI - A gender approach to work ability and its relationship to professional and domestic work hours among nursing personnel. AB - The association between working hours and work ability was examined in a cross sectional study of male (N=156) and female (N=1092) nurses in three public hospitals. Working hours were considered in terms of their professional and domestic hours per week and their combined impact; total work load. Logistic regression analysis showed a significant association between total work load and inadequate work ability index (WAI) for females only. Females reported a higher proportion of inadequate WAI, fewer professional work hours but longer domestic work hours. There were no significant differences in total work load by gender. The combination of professional and domestic work hours in females seemed to best explain their lower work ability. The findings suggest that investigations into female well-being need to consider their total work load. Our male sample may have lacked sufficient power to detect a relationship between working hours and work ability. PMID- 18405879 TI - Histochemistry of nerve fibres double labelled with anti-TRPV2 antibodies and sensory nerve marker AM1-43 in the dental pulp of rat molars. AB - AM1-43 can label sensory nerve fibres and sensory neurons. Permeation of non selective cation channels of the nerve cell membrane is suggested to be the mechanism responsible for labelling. To identify these channels, two candidates, TRPV1 and TRPV2 were examined by immunocytochemistry in the dental pulp and trigeminal ganglion of rats injected with AM1-43. A part of AM1-43-labelled nerve fibres was also positive for anti-TRPV2 antibody but negative for anti-TRPV1 antibody in the dental pulp. In the trigeminal ganglion, a part of the neuron showed both bright AM1-43 labelling and anti-TRPV2 immunolabelling, but neurons double labelled with AM1-43 and TRPV1 were rare. These results suggest that TRPV2 channels, but not TRPV1 channels, contribute to the fluorescent labelling of AM1 43 in the dental pulp. PMID- 18405880 TI - The influence of data-driven processing on perceptions of memory quality and intrusive symptoms in children following traumatic events. AB - Ehlers and Clark [(2000). A cognitive model of post-traumatic stress disorder. Behaviour Research and Therapy, 38, 319-345] cognitive model of post-traumatic stress disorder (PTSD) has been relatively untested with children. Seventy-five children (7-16 years) were interviewed within 4 weeks of an injury that led to hospital treatment to examine whether peri-traumatic processing strategies (data driven processing and fear) were associated with perceptions of memory quality and intrusive memories. Perceptions of memory quality mediated the relationship between data-driven processing and intrusive reactions but not avoidance, arousal or depressive reactions. Finally, the relationship between peri-event fear and intrusion reactions was mediated by perceptions of memory quality even after data driven processing was controlled. The implications of these findings are discussed in the context of a cognitive developmental model of PTSD in children. PMID- 18405881 TI - Treatment of lung cancer cells with cytotoxic levels of sodium selenite: effects on the thioredoxin system. AB - Selenium at subtoxic doses has been shown to have tumor specific cytotoxic effects. In this work, viability measurements in different lung cancer cell lines showed that selenite was more effective compared to three different conventional cytotoxic drugs. In addition, the cell line most sensitive to selenite toxicity comprised the highest level of thioredoxin reductase 1 (TrxR1). The human selenoenzyme TrxR1 is a central enzyme for cell growth, differentiation, and the protection against oxidative stress. TrxR1, which in several studies has been shown to be up-regulated in various tumor cells, is also a target for many anticancer drugs. In this study, inhibition of TrxR resulted in enhanced selenite cytotoxicity, clearly connecting the thioredoxin system to the toxic effects mediated by selenite. The complex regulation of TrxR1, involving the expression of many different transcript forms of mRNA, was investigated by real-time qPCR in lung cancer cell lines following treatment with toxic doses (2.5-10 microM) of sodium selenite. Selenium treatment resulted in increased expression of almost all TrxR1 mRNA variants with increasing concentrations of selenite. On the contrary, the TrxR protein level and activity, increased at low to moderate doses followed by a decrease at higher doses, indicating impairment of protein synthesis by selenite. PMID- 18405882 TI - Heritability of sleep electroencephalogram. AB - BACKGROUND: Understanding the basis of sleep-related endophenotypes might help to pinpoint factors modulating susceptibility to psychiatric disorders. However, the genetic underpinnings of sleep microarchitecture in humans remain largely unknown. Here we report on the results of a classical twin study in monozygotic (MZ) and dizygotic (DZ) twin pairs examining the genetic effect on sleep electroencephalogram (EEG) composition. METHODS: Polysomnographic recordings were obtained in 35 pairs of MZ (26.4 +/- 5.4 years, 17-43 years, 17 male pairs, 18 female pairs) and 14 same-gender pairs of DZ twins (22.1 +/- 2.7 years, 18-26 years, 7 male pairs, 7 female pairs). The EEG power spectra were generated on the basis of Fast Fourier transformations combined with conventional sleep parameters, according to standardized criteria. RESULTS: We tested the genetic variance contributing to the observed overall variance of the sleep measures and found that the relative contributions of the delta, theta, alpha, and sigma frequency bands at central derivations were significantly correlated to the genetic background. In these frequency bands, MZ twins also showed within-pair concordance in spectral power that was significantly higher than that of DZ twins. CONCLUSIONS: The broad overlap of EEG frequencies during non-REM sleep and wakefulness, which shows a significant genetic variance, supports the hypothesis of common neuronal mechanisms generating EEG oscillations in humans. Our findings strongly support the suitability of the spectral composition of non-REM sleep for defining endophenotypes. PMID- 18405883 TI - Antistress effect of TRPV1 channel on synaptic plasticity and spatial memory. AB - BACKGROUND: Stress is believed to exacerbate neuropsychiatric and cognitive disorders. In particular, the hippocampus, which plays critical roles in certain types of memory, including spatial memory, is exquisitely sensitive to stress. Certain types of memory are believed to depend on activity-dependent hippocampal synaptic plasticity such as long-term potentiation (LTP) and long-term depression (LTD), but stress suppresses LTP and facilitates LTD in the hippocampus and impairs spatial memory. Although the transient receptor potential vanilloid 1 (TRPV1 or VR1) is widely expressed in the hippocampus, it remains unknown whether the TRPV1 channel antagonizes the stress effects on hippocampal function. METHODS: Using the TRPV1 agonists capsaicin and resiniferatoxin and selective antagonists capsazepine and SB366791, we examined the effect of TRPV1 activation on LTP and LTD in hippocampal CA1 slices of juvenile rats. Furthermore, we examined whether the effects of acute stress on synaptic plasticity and spatial memory could be prevented by intrahippocampal or intragastric infusion of a TRPV1 agonist. RESULTS: The TRPV1 agonists capsaicin and resiniferatoxin facilitated LTP but suppressed LTD. Alterations were mediated by TRPV1 because the TRPV1 selective antagonists capsazepine and SB366791 blocked the actions of capsaicin. Acute stress suppressed LTP and enabled LTD, but the TRPV1 agonist capsaicin effectively prevented this effect. When capsaicin was intrahippocampally or intragastrically infused, the acute stress effect on impairing spatial memory retrieval was completely prevented. CONCLUSIONS: The TRPV1 channel is a potential target to facilitate LTP and suppress LTD, in turn protecting hippocampal synaptic plasticity and spatial memory retrieval from the influence of acute stress. PMID- 18405884 TI - Graft of pre-injured sural nerve promotes regeneration of corticospinal tract and functional recovery in rats with chronic spinal cord injury. AB - A possible treatment approach for chronic spinal cord injuries has been tested. We report that minced, autologous, pre-injured peripheral nerve administered as a single injection into an injury-induced cyst, resulting from a contusion injury of the thoracic spinal cord, stimulates recovery of hindlimb locomotor function in rats, as measured by the Basso, Beattie, Bresnahan Locomotor Rating Scale. This response was further enhanced by the addition of exogenous neurotrophic factors. Histological analysis showed axons of the corticospinal tract exhibited significant regeneration past the injury site, when quantified both by number and length. Results indicate that the use of a pre-injured peripheral nerve graft stimulates chronically injured descending nerves to overcome a local inhibitory environment. The resulting sprouting and growth past the injury site is associated with a significant improvement in locomotor function. PMID- 18405885 TI - Neurochemical architecture of the filum terminale in the rat. AB - Contrary to the widespread assumption, the filum terminale in the rat possesses a precise glial and neuronal organization. The processes of glial fibrillary acidic protein-stained astrocytes form a rich, three dimensional array. The crescent shaped white matter could be outlined with antibody detecting oligodendrocytes. The neurons in the filum terminale, labeled with neuron-specific nuclear protein, are distributed in a small midline group (dorsal nucleus) dorsal to and in two symmetrical clusters at both sides of the central canal (lateral nuclei). Nitric oxide synthase-, calretinin-, choline acetyltransferase-, substance P- and neurokinin receptor-1-immunoreactive neurons were detected in the lateral nuclei. Axons were classified based on their course and termination. Small number of calcitonin gene-related peptide-immunoreactive fibers was found exclusively in the dorsal nucleus. Nitric oxide synthase-, substance P-, and neurokinin receptor 1-stained axon arborizations were detected mainly in the lateral nucleus. A dense array of extremely fine vesicular glutamate transporter 2- and fine, synaptophysin-immunoreactive varicosities covered densely the lateral nuclei. Fine glycine-transporter 2-immunoreactive axon arborization like structures were seen also in the lateral nucleus. Vesicular glutamate transporter 1- and choline acetyltransferase-immunoreactive axons arborized in the entire gray matter. Serotonin- and enkephalin-immunoreactive fibers congregated in the dorsolateral portion of the white matter, called "shoulder region", while calretinin- and thick, varicose neurokinin receptor-1-stained axons were also seen in the same area of the white matter. Synaptophysin-immunoreactive fine varicosities colocalized only with vesicular glutamate transporter 2 immunoreaction. Substance P and glycine-transporter 2-immunoreactive puncta were found in close contact with neurokinin receptor-1-immunostained perikarya and dendrites. PMID- 18405886 TI - Selenium protects the hypoxia induced apoptosis in neuroblastoma cells through upregulation of Bcl-2. AB - Selenium (Se) is an essential micronutrient as well as a toxic trace element in animal and human nutrition. The effects of Se in the immune system and some diseases are well documented. The objective of the present study was to examine the role of Se in reducing the hypoxia induced apoptosis in neuroblastoma cell line. Hypoxia showed an enhanced cytotoxicity, increased free radical production and apoptosis (p<0.001) which was measured in terms of DNA break down by comet assay. Hypoxia has decreased reduced Glutathione (GSH) content, Glutathione Reductase (GR), Glutathione peroxidase (GPx) and Superoxide Dismutase (SOD) activities as compared to control cells. During hypoxic condition the expression of cytochrome C, pro and active caspase-3 levels were enhanced significantly followed by nonsignificant upregulation of Bcl-2. But, the Se supplementation inhibited the cytotoxicity, free radical generation and stabilized the HIF-1alpha accumulation in cells under hypoxia. The GSH content, GR, GPx and SOD activities increased significantly in Se-treated hypoxic cells, as compared to control. Further there was an appreciable inhibition of apoptosis by upregulation of Bcl-2 proteins, in the presence of Se under hypoxia. Selenium supplementation to cells significantly inhibited the hypoxia induced DNA fragmentation and restored the antioxidant status back to control levels. This study suggests that Se supplementation prevented the cells from hypoxia induced apoptosis by triggering upregulation of Bcl-2 protein and reducing the oxidative stress. PMID- 18405888 TI - Unusual addition of amines to C-2 of vinyl sulfone-modified-beta-D-pent-2 enofuranosyl carbohydrates: synthesis of a new class of beta-anomeric 2-amino-2,3 dideoxy-D-threo-pentofuranosides. AB - When 3-C-sulfonyl-pent-2-enofuranosides and 3-C-sulfonyl-hex-2-enofuranosides were reacted with primary and secondary amines, only the beta-anomeric methoxy group of the pent-2-enofuranoside did not cause any hindrance to incoming nitrogen nucleophiles. This resulted in the 'unusual' addition of amines, in which the diastereoselectivity of the reaction was overwhelmingly in favor of amino sugars of the D-arabino configuration. Selected products were desulfonylated to obtain a new class of beta-anomeric 2-amino-2,3-dideoxy-D-threo pentofuranosides. PMID- 18405887 TI - Chemoenzymatic synthesis of glycopeptides and glycoproteins through endoglycosidase-catalyzed transglycosylation. AB - Homogeneous glycopeptides and glycoproteins are indispensable for detailed structural and functional studies of glycoproteins. It is also fundamentally important to correct glycosylation patterns for developing effective glycoprotein based therapeutics. This review discusses a useful chemoenzymatic method that takes advantage of the endoglycosidase-catalyzed transglycosylation to attach an intact oligosaccharide to a polypeptide in a single step, without the need for any protecting groups. The exploration of sugar oxazolines (enzymatic reaction intermediates) as donor substrates has not only expanded substrate availability, but also has significantly enhanced the enzymatic transglycosylation efficiency. Moreover, the discovery of a novel mutant with glycosynthase-like activity has made it possible to synthesize homogeneous glycoproteins with full-size natural N glycans. Recent advances in this highly convergent chemoenzymatic approach and its application for glycopeptide and glycoprotein synthesis are highlighted. PMID- 18405889 TI - Interleukin-10 containing normal human serum inhibits granzyme B release but not perforin release from alloreactive and EBV-specific T cell clones. AB - Interleukin-10 (IL-10), also known as cytokine synthesis inhibitory factor, is capable of inhibiting synthesis of pro-inflammatory cytokines like IFNgamma, IL 2, IL-3, TNFalpha and GM-CSF made by cells such as macrophages and T helper Type 1 cells. We observed that normal human serum, derived from a healthy individual but containing large amounts of IL-10, inhibited cytotoxic activity and interfered with granzyme B release from alloreactive cytotoxic T cell (CTL) clones in vitro, but did not affect perforin release. The addition of normal human serum containing high levels of anti-IL-10 IgG neutralized the inhibitory effects of IL-10 serum. Moreover, we have identified that cytotoxic activity and granzyme B release from an Epstein-Barr virus (EBV)-specific CTL clone was similarly inhibited in the presence of IL-10 serum, while perforin release was unaffected. Anti-IL-10 IgG serum also appeared to neutralize the inhibitory effect of IL-10 serum on an EBV-specific CTL clone. PMID- 18405890 TI - Immune up regulatory response of a non-caloric natural sweetener, stevioside. AB - Immunomodulation is a process, which alters the immune system of an organism by interfering with its functions. This interference results in either immunostimulation or immunosuppression. An immunomodulator is any substance that helps to regulate the immune system. This "regulation" is a normalization process, so that an immunomodulator helps to optimise immune response. Immunomodulators are becoming very popular in the worldwide natural health industry as these do not tend to boost immunity, but to normalize it. Keeping this in view, major efforts have to be directed to modulate the immune responses, to permit effective treatment of various ailments associated with immune system and thus the development of a safe and effective immunomodulator for clinical us. Leaves of Stevia rebaudiana are a source of several sweet glycosides of steviol. The major glycoside, stevioside, diterpenoid glycoside--is used in oriental countries as a food sweetener. Its medical use is also reported as a heart tonic. Besides, it is used against obesity, hypertension, and stomach burn and to lower uric acid levels. Here in this study, stevioside was tested for its immunomodulatory activity on different parameters of the immune system at three different doses (6.25, 12.5 and 25 mg/kg p.o.) on normal as well as cyclophosphamide treated mice. Stevioside was found effective in increasing phagocytic activity, haemagglutination antibody titre and delayed type hypersensitivity. In parallel, stevioside substantially increase proliferation in the LPS and Con A stimulated B and T cells, respectively. Present study, therefore, reveals that the drug holds promise as immunomodulating agent, which acts by stimulating both humoral as well as cellular immunity and phagocytic function. PMID- 18405891 TI - Efficacy of caffeic acid in preventing nickel induced oxidative damage in liver of rats. AB - Nickel (Ni), a major environmental pollutant, is known for its wide toxic manifestations. In the present study caffeic acid (CA), one of the most commonly occurring phenolic acids in fruits, grains and dietary supplements, was evaluated for its protective effect against the Ni induced oxidative damage in liver. In this investigation, Ni (20 mg/kg body weight) was administered intraperitoneally for 20 days to induce toxicity. CA was administered orally (15, 30 and 60 mg/kg body weight) for 20 days with intraperitoneal administration of Ni. Ni induced liver damage was clearly shown by the increased activities of serum hepatic enzymes namely aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT) and lactate dehydrogenase (LDH) along with increased elevation of lipid peroxidation indices (thiobarbituric reactive acid substances (TBARS) and lipid hydroperoxides). The toxic effect of Ni was also indicated by significantly decreased levels of enzymatic (superoxide dismutase (SOD), catalase (CAT) glutathione peroxidase (GPx) and glutathione S-transferase (GST)) and non-enzymatic antioxidants (glutathione (GSH), vitamin C and vitamin E). CA administered at a dose of 60 mg/kg body weight significantly reversed the activities of hepatic marker enzymes to their near normal levels when compared with other two doses. In addition, CA significantly reduced lipid peroxidation and restored the levels of antioxidant defense in the liver. All these changes were supported by histological observations. The results indicate that CA may be beneficial in ameliorating the Ni induced oxidative damage in the liver of rats. PMID- 18405892 TI - Flatworm stem cells and the germ line: developmental and evolutionary implications of macvasa expression in Macrostomum lignano. AB - We have isolated and identified the vasa homologue macvasa, expressed in testes, ovaries, eggs and somatic stem cells of the flatworm Macrostomum lignano. Molecular tools such as in situ hybridization and RNA interference were developed for M. lignano to study gene expression and function. Macvasa expression was followed during postembryonic development, regeneration and in starvation experiments. We were able to follow gonad formation in juveniles and the reformation of gonads from stem cells after amputation by in situ hybridization and a specific Macvasa antibody. Expression of macvasa in the germ cells was highly affected by feeding conditions and correlated with the decrease and regrowth of the gonads. RNA interference showed specific down-regulation of macvasa mRNA and protein. The absence of Macvasa did not influence gonad formation and stem cell proliferation. Our results corroborate the exclusive nature of the flatworm stem cell system but challenge the concept of a solely postembryonic specification of the germ line in Platyhelminthes. We address the transition of somatic stem cells to germ cells and speculate on Macrostomum as a system to unravel the mechanisms of preformation or epigenesis in the evolution of germ line specification from somatic stem cells. PMID- 18405893 TI - Dimethyl fumarate, a small molecule drug for psoriasis, inhibits Nuclear Factor kappaB and reduces myocardial infarct size in rats. AB - Persistent Nuclear Factor-kappaB (NF-kappaB) activation is hypothesized to contribute to myocardial injuries following ischemia-reperfusion. Because inhibition or control of NF-kappaB signaling in the heart probably confers cardioprotection, we determined the potency of the NF-kappaB inhibitor dimethyl fumarate (DMF) in cardiovascular cells, and determined whether administration of DMF translates into beneficial effects in an animal model of myocardial infarction. In rat heart endothelial cells (RHEC), we analysed inhibitory effects of DMF on NF-kappaB using shift assay and immunohistofluorescence. In in vivo experiments, male Sprague Dawley rats undergoing left coronary artery occlusion for 45 min received either DMF (10 mg/kg body weight) or vehicle 90 min before ischemia as well as immediately before ischemia. After 120 min of reperfusion, the hearts were stained with phthalocyanine blue dye and triphenyltetrazolium chloride. Additionally, acute hemodynamic and electrophysiologic effects of DMF were determined in dose-response experiments in isolated perfused rat hearts. DMF inhibited TNF-alpha-induced nuclear entry of NF-kappaB in RHEC. In in vivo experiments, myocardial infarct size was significantly smaller in rats that had received DMF (20.7%+/-9.7% in % of risk area; n=17) than in control rats (28.2%+/ 6.2%; n=15). Dose-response experiments in isolated perfused rat hearts excluded acute hemodynamic or electrophysiologic effects as mechanisms for the effects of DMF. DMF inhibits nuclear entry of NF-kappaB in RHEC and reduces myocardial infarct size after ischemia and reperfusion in rats in vivo. There was no indication that the beneficial effects of DMF were due to acute hemodynamic or electrophysiologic influences. PMID- 18405895 TI - The role of the lens in refractive development of the eye: animal models of ametropia. AB - Research with young mammals and chicks has shown that the visual environment can affect the refractive development of the eye by enhancing or slowing axial eye growth, but the effect on the refractive components of the eye, the lens and cornea, are less clear. A review of the literature indicates that the lens is minimally affected, if at all, and results vary depending on whether the lens is studied in an isolated state or with the accommodative apparatus intact. Research has shown that the development of myopia or hyperopia in young chicks alters lens focal length and magnitude of the accommodative response. However, the result may be indirect or passive due to the effect of the change in size and shape of the globe on the articulation between the ciliary body and lens. Recent research has also investigated the role of the lens in induced refractive error development in a fish, tilapia (Oreochromis niloticus). Translucent goggles were sutured over one eye for 4 weeks to induce form deprivation myopia while the untreated eye served as an untreated contralateral control. In addition to measuring refractive state and intraocular dimensions, a scanning laser system was used to determine the optical quality of excised lenses. All the deprived fish eyes developed significant amounts of myopia and the vitreous and anterior chambers of the treated eye were significantly longer axially than those of the untreated contralateral eyes. No significant change in optical quality was found between lenses of the myopic and non-myopic eyes and the fish recovered completely from the myopia five days after the goggle was removed. The results show that although fish, unlike higher vertebrates, are capable of lifelong growth, the visual environment is an important factor controlling ocular development in this group as well, and eye development is not strictly genetically determined. This review indicates that lens growth and optical development is independent from the refractive development of the whole eye. PMID- 18405896 TI - Expression and localisation of BDNF, NT4 and TrkB in proliferative vitreoretinopathy. AB - Exogenous brain derived neurotrophic factor (BDNF) is known to rescue ganglion cell death after optic nerve injury. Its mechanism of action is believed to be indirect via glial cells in the retina. In this study we investigated the changes in expression and localisation of BDNF, neurotrophin-4 (NT4) and their common receptor (TrkB) in retinectomy sections of patients with proliferative vitreoretinopathy (PVR). Nine full-thickness retinectomy specimens obtained at retinal reattachment surgery for PVR were fixed in 4% paraformaldehyde immediately after excision and compared to similarly processed normal donor retinas (4 eyes). Agarose-embedded sections (100 microm thick) were double labelled for immunohistochemistry by confocal microscopy, with antibodies against BDNF, NT4, TrkB, rod opsin, glial fibrillary acidic protein (GFAP), cellular retinaldehyde binding protein (CRALBP) and Brn3. This study demonstrates expression of NT4 by ganglion cells and shows expression of BDNF and NT4 in the outer photoreceptor segments is downregulated during PVR, whilst NT4 is markedly upregulated throughout the retina during this condition. The findings here suggest that NT4 may play a neural protective role during the development of PVR. It also shows that upregulation of NT4 in PVR is localised to Muller glial cells, indicating either over-expression of this factor by Muller cells or that Muller cells internalise NT4 for trafficking across the retina. TrkB expression was not observed in PVR retina. The observations that Muller glia demonstrate upregulation of NT4 suggests that retinal injury may lead to activation of this neurotrophin by Muller cells as part of their neuroprotective functions. PMID- 18405894 TI - Evidence for AMPK-dependent regulation of exocytosis of lipoproteins in a model liver cell line. AB - 5'-AMP-activated kinase (AMPK) plays a key role in the regulation of cellular lipid metabolism. The contribution of vesicular exocytosis to this regulation is not known. Accordingly, we studied the effects of AMPK on exocytosis and intracellular lipid content in a model liver cell line. Activation of AMPK by metformin or 5-aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) increased the rates of constitutive exocytosis by about 2-fold. Stimulation of exocytosis by AMPK occurred within minutes, and persisted after overnight exposure to metformin or AICAR. Activation of AMPK also increased the amount of triacylglycerol (TG) and apolipoprotein B (apoB) secreted from lipid-loaded cells. These effects were accompanied by a decrease in the intracellular lipid content indicating that exocytosis of lipoproteins was involved in these lipid lowering effects. While AMPK increased the rates of fatty acid oxidation (FAO), the lipid-lowering effects were quantitatively significant even after inhibition of FAO with R-etomoxir. These results suggest that hepatic AMPK stimulates constitutive exocytosis of lipoproteins, which may function in parallel with FAO to regulate intracellular lipid content. PMID- 18405897 TI - Hemorrhagic emphysematous cholecystitis presenting as obstructive jaundice and hemobilia. PMID- 18405898 TI - Central cardiovascular actions of angiotensin II in trout. AB - In mammals, a large body of evidence supports the existence of a brain renin angiotensin system (RAS) acting independently or synergistically with the endocrine RAS to maintain diverse physiological functions, notably cardiovascular homeostasis. The RAS is of ancient origin and although most components of the RAS are present within the brain of teleost fishes, little is known regarding the central physiological actions of the RAS in these vertebrates. The present review encompasses the most relevant functional data for a role of the brain RAS in cardiovascular regulations in our experimental animal model, the unanesthetized trout Oncorhynchus mykiss. This paper mainly focuses on the central effect of angiotensin II (ANG II) on heart rate, blood pressure, heart rate variability and cardiac baroreflex, after intracerebroventricular injection or local microinjection of the peptide within the dorsal vagal motor nucleus. The probable implications of the parasympathetic nervous system in ANG II-evoked changes in the cardiac responses are also discussed. PMID- 18405900 TI - Cigarette smoke is an endothelial stressor and leads to cell cycle arrest. AB - The molecular mechanisms underlying the atherogenic activity of cigarette smoke have yet to be fully elucidated. In the present study, genome-wide microarray analysis was performed on endothelial cells exposed to an aqueous cigarette smoke extract (CSE) for 3, 7, and 24 h, to obtain a better insight into how smoking may lead to endothelial damage. Microarray analysis showed the transcriptional response to CSE was dominated by heat shock, stress responsive, and inflammatory genes, along with genes encoding for anti-oxidant and metal detoxification proteins. The heat shock response was shown to be a result of short lived reactive species of CSE, with the abrogation of the effect by the addition of old CSE, the anti-oxidant N-acetyl cysteine, or the removal of metals from CSE implying that reactive oxygen species are the main culprit. This was further supported by a strong decline in the level of intracellular protein oxidation levels seen under these conditions compared to freshly prepared CSE. Mitochondrial integrity was also found to be significantly compromised after CSE treatment, resulting in a threefold increase in depolarised mitochondria after 6 h. Finally, cell cycle analysis showed the induction of G1 cell cycle arrest. An increased stress and inflammation response indicates that endothelial damage from smoking could contribute to immune cell infiltration, while decreased growth rates reduce endothelial layer repair, promoting atherogenesis. PMID- 18405899 TI - Exogenous kisspeptin does not alter photoperiod-induced gonadal regression in Siberian hamsters (Phodopus sungorus). AB - In order to reproduce successfully, animals must integrate multiple environmental cues to synchronize breeding with favorable conditions. In temperate, seasonally breeding rodents, photoperiod acts as the primary seasonal cue. Long days are associated with reproductive development and maturation of the gonads whereas short days induce gonadal regression. The neuropeptide kisspeptin has potent stimulatory effects on reproductive development. Kisspeptin potently stimulates GnRH release and kisspeptin expression co-varies with photoperiod in seasonally breeding animals. Here we tested the hypothesis that reproductive involution in response to inhibitory day lengths results from reduced kisspeptin stimulation of the reproductive axis in seasonally breeding Siberian hamsters (Phodopus sungorus). If true, gonadal regrowth should be hastened by kisspeptin treatment in regressed hamsters and prevented in hamsters by treatment prior to and during regression. In Experiments 1 and 2 we tested the ability of kisspeptin to reverse gonadal regression. In Experiment 1, reproductively regressed hamsters received chronic kisspeptin via osmotic mini-pumps for 4 weeks. In Experiment 2, daily injections of kisspeptin were administered to regressed hamsters for 6 weeks. In Experiment 3, the ability of kisspeptin to block gonadal regression was tested; hamsters transferred to short days received daily injections of kisspeptin for 6 weeks. In all three studies, short-day animals receiving exogenous kisspeptin did not differ from short-day controls. Collectively, these results provide evidence that mechanisms in addition to those that converge on the kisspeptin system are likely critical for seasonal changes in the reproductive axis. PMID- 18405901 TI - Cardiovascular risk-factors predict progression of urinary albumin-excretion in a general, non-diabetic population: a gender-specific follow-up study. AB - Increased urinary albumin-excretion (UAE) predicts cardiovascular events and clusters with the metabolic syndrome. The aim of this population-based, prospective study was to assess the relationship between baseline and longitudinal changes in cardiovascular risk-factors and 7 years' increase in UAE. Three thousand and four hundred non-diabetic participants (1838 men, 1562 women) of the Tromso studies in 1994/1995 and 2001/2002 were included. In each survey, first-void spot-urine-samples were collected, and albumin-creatinine ratio (ACR) was calculated. Change in ACR (DeltaACR) was dichotomized into upper vs. the three lower quartiles. Median UAE in the population did not increase during follow-up. Baseline predictors for DeltaACR in the upper quartile were: age (OR 1.32 per 5 years, 95% CI 1.22-1.43), HbA1c (OR 1.43 per %, 95% CI 1.08-1.91) and waist circumference (OR 1.11 per 5 cm, 95% CI 1.04-1.19) in men, and age (OR 1.14 per 5 years, 95% CI 1.04-1.25) and current smoking (OR 1.71, 95% CI 1.27-2.30) in women. Systolic blood pressure and estimated glomerular filtration rate were predictors without gender-specificity. Clustering of three or more metabolic traits did not predict ACR increase independently. Protective factors against ACR increase were initiation of antihypertensive treatment in women (OR 0.59, 95% CI 0.39-0.87) and hard physical activity in men (OR 0.70, 95% CI 0.51-0.96). In summary, cardiovascular risk-factors at baseline predicted ACR increase, but initiation of antihypertensive therapy (women) and physical activity (men) seemed to protect from ACR increase during follow-up. Endpoint-data are needed to explore the clinical significance of low-grade UAE increase. PMID- 18405902 TI - Local dynamic stability in turning and straight-line gait. AB - Successful community and household ambulation require the ability to navigate corners and maneuver around obstacles, posing unique challenges compared to straight-line walking. The challenges associated with turning may contribute to an increased incidence of falling and the occurrence of fall-related injuries. A measure of stability applied to turning gait may be able to quantify a system's response to naturally occurring disturbances associated with turning and identify subjects at greater risk of falling. An index of stability has been used previously to assess the rate of kinematic separation (local dynamic stability) during straight-line gait. The purpose of this study was to determine if local dynamic stability during constant speed turning is reduced compared to straight line treadmill walking. Maximum finite-time Lyapunov exponents (lambda) were used to estimate the local stability of able-bodied subjects' (n=19) sagittal plane hip, knee, and ankle trajectories for turning compared to straight-line walking at two different walking speeds. Turning lambda was greater than straight lambda for the hip, right knee, and ankle (p<0.05). Turning lambda for the left knee angle was similar to straight lambda. There were no differences in lambda between left and right limbs for the hip and ankle and also no differences between the inside and outside limbs during turning for all joints. These findings indicate able-bodied subjects' hip, right knee, and ankle kinematics are less locally stable while turning than walking in a straight line and may be used as a comparative tool for determining the efficacy of therapeutic interventions for mobility-impaired populations. PMID- 18405903 TI - Determination of instantaneous stability against backward balance loss: two computational approaches. AB - The purposes of this study were (1) to develop two computational methods and (2) to compare their performance (i.e. the accuracy and the time demand) in determining instantaneous stability, i.e., the shortest distance from an instantaneous center of mass (COM) state to all interpolated points on the stability boundary in the COM position-velocity state space. The intuitive numerical method interpolated the stability boundary against backward balance loss based on four sets of discrete boundary values with a spline function and a preset step size (Deltax). Then a brute-force search approach was used to determine this shortest distance. The analytical method used quadratic polynomial functions to represent the boundaries. The stability was then determined by using the Lagrange multiplier method to find the optimum for the analytical expression of the distance from this given COM state to the boundary. Because reducing Deltax can supposedly improve the accuracy of spline fitting, we used this approach to establish the benchmark employed to determine the accuracy level of all other computations. Based on the results from a database of 67 sit-to-stand slip trials and 687 gait-slip trials, we would recommend the numerical method with Deltax=0.1 to reach an adequate accuracy level of 10(-2). By changing one order of Deltax from 0.1 to 0.01, however, the accuracy level improved drastically from 10(-2) to 10(-4). Nonetheless to achieve the same higher accuracy at a faster computing speed, we would recommend the analytical method, which costs 80% less time than the numerical approach at the accuracy level of 10(-4). PMID- 18405904 TI - In-vivo range of motion of the subtalar joint using computed tomography. AB - Understanding in vivo subtalar joint kinematics is important for evaluation of subtalar joint instability, the design of a subtalar prosthesis and for analysing surgical procedures of the ankle and hindfoot. No accurate data are available on the normal range of subtalar joint motion. The purpose of this study was to introduce a method that enables the quantification of the extremes of the range of motion of the subtalar joint in a loaded state using multidetector computed tomography (CT) imaging. In 20 subjects, an external load was applied to a footplate and forced the otherwise unconstrained foot in eight extreme positions. These extreme positions were foot dorsiflexion, plantarflexion, eversion, inversion and four extreme positions in between the before mentioned positions. CT images were acquired in a neutral foot position and each extreme position separately. After bone segmentation and contour matching of the CT data sets, the helical axes were determined for the motion of the calcaneus relative to the talus between four pairs of opposite extreme foot positions. The helical axis was represented in a coordinate system based on the geometric principal axes of the subjects' talus. The greatest relative motion between the calcaneus and the talus was calculated for foot motion from extreme eversion to extreme inversion (mean rotation about the helical axis of 37.3+/-5.9 degrees, mean translation of 2.3+/ 1.1 mm). A consistent pattern of range of subtalar joint motion was found for motion of the foot with a considerable eversion and inversion component. PMID- 18405905 TI - Construction of an automated gas chromatography/mass spectrometry system for the analysis of ambient volatile organic compounds with on-line internal standard calibration. AB - An automated sampling and enrichment apparatus coupled with a gas chromatography/mass spectrometry (GC/MS) technique was constructed for the analysis of ambient volatile organic compounds (VOCs). A sorbent trap was built within the system to perform on-line enrichment and thermal desorption of VOCs onto GC/MS. In order to improve analytical precision, calibration accuracy, and to safe-guard the long-term stability of this system, a mechanism to allow on line internal standard (I.S.) addition to the air sample stream was configured within the sampling and enrichment apparatus. A sub-ppm (v/v) level standard gas mixture containing 1,4-fluorobenzene, chloropentafluorobenzene, 1-bromo-4 fluorobenzene was prepared from their pure forms. A minute amount of this I.S. gas was volumetrically mixed into the sample stream at the time of on-line enrichment of the air sample to compensate for measurement uncertainties. To assess the performance of this VOC GC/MS system, a gas mixture containing numerous VOCs at sub-ppb (v/v) level served as the ambient air sample. Various internal standard methods based on total ion count (TIC) and selective ion monitoring (SIM) modes were attempted to assess the improvement in analytical precision and accuracy. Precision was improved from 7-8% RSD without I.S. to 2-3% with I.S. for the 14 target VOCs. Uncertainties in the calibration curves were also improved with the adoption of I.S. by reducing the relative standard deviation of the slope (Sm%) by an average a factor of 4, and intercept (Sb%) by a factor of 2 for the 14 target VOCs. PMID- 18405906 TI - Separation of organic cations using novel background electrolytes by capillary electrophoresis. AB - A background electrolyte for capillary electrophoresis containing tris( hydroxymethyl) aminomethane (THAM) and ethanesulfonic acid (ESA) gives excellent efficiency for separation of drug cations with actual theoretical plate numbers as high as 300,000. However, the analyte cations often elute too quickly and consequently offer only a narrow window for separation. The best way to correct this is to induce a reverse electroosmotic flow (EOF) that will spread out the peaks by slowing their migration rates, but this has always been difficult to accomplish in a controlled manner. A new method for producing a variable EOF is described in which a low variable concentration of tributylammonium- or triethylammonium ESA is added to the BGE. The additive equilibrates with the capillary wall to give it a positive charge and thereby produce a controlled opposing EOF. Excellent separations of complex drug mixtures were obtained by this method. PMID- 18405907 TI - Microwave-assisted derivatization of acidic herbicides for gas chromatography mass spectrometry. AB - Microwave radiation is used to speed up chemical derivatization. In the present study, three microwave-assisted techniques for the methylation of chlorophenoxy acid herbicides prior to analysis by gas chromatography coupled with mass spectrometry are compared. Derivatization was performed with the catalysts sulphuric acid and boron trifluoride as well as with trimethylsilyldiazomethane. In order to establish optimized and stable conditions, a screening for statistically significant factors by means of experimental designs was carried out and supplemented by a careful optimization. Special emphasis has been given to an accurate validation to prove the performance of the techniques. Furthermore, all microwave-assisted methods were compared with their conventional analogues. The optimized methods are valid for routine analysis of different matrices such as water, soil, sediment or tissues, especially for high sample throughput since a simultaneous derivatization of up to 64 samples in one run is possible. PMID- 18405908 TI - Phenomena of insulin peak fronting in size exclusion chromatography and strategies to reduce fronting. AB - Insulin peak fronting in size exclusion chromatography (SEC) results in more than 10% yield loss in the production of insulin. The goal of this study is to understand the mechanisms of peak fronting and to develop strategies to reduce fronting and increase insulin yield. Chromatography experiments ruled out pressure surge, viscous fingering, and adsorption as the cause for peak fronting. Theoretical analysis based on a general rate model indicated that reversible dimerization is the major cause for peak fronting and reducing the dimerization equilibrium constant is the most effective method for reducing fronting. Two strategies were developed and tested to reduce the degree of dimer formation. The first strategy was to use 0.1N acetic acid as the presaturant and eluent. The second strategy was to use 0.8 or 2.8N acetic acid in 20vol.% denatured ethanol as the mobile phase. The experimental results showed that both strategies can reduce insulin peak fronting in SEC, maintain desired product purity, and increase insulin yield to higher than 98%. PMID- 18405909 TI - Analysis of polybrominated diphenyl ethers in atmospheric deposition and snow samples by solid-phase disk extraction. AB - An extraction method for the quantitative analysis of polybrominated diphenyl ethers (PBDEs) in aqueous samples has been evaluated. The analytical methodology includes the sample filtration through glass fiber filter and subsequent extraction of dissolved phase compounds by C18 solid-phase disk extraction. Dependence of extraction efficiency on factors such as pollutant concentrations, sample volume, and stability during storage has been investigated. Mean extraction efficiencies of 97% for total PBDEs (13 tri- to heptabrominated congeners at spiking levels in the range of 15-90pg) with a RSD between 9 and 20% were achieved. Higher recoveries were observed for the more volatile PBDEs (112%) in relation to more brominated congeners (88%). The developed methodology was successfully applied to the analysis of these compounds in atmospheric deposition and snow samples from remote sites in Europe with method detection and quantitation limits in the range of 2.1-10pgL(-1) for almost all congeners, which allow the determination of PBDEs in remote areas with levels in the range of low to medium pgL(-1) for SigmaPBDEs. PMID- 18405910 TI - Interaction forces between waterborne bacteria and activated carbon particles. AB - Activated carbons remove waterborne bacteria from potable water systems through attractive Lifshitz-van der Waals forces despite electrostatic repulsion between negatively charged cells and carbon surfaces. In this paper we quantify the interaction forces between bacteria with negatively and positively charged, mesoporous wood-based carbons, as well as with a microporous coconut carbon. To this end, we glued carbon particles to the cantilever of an atomic force microscope and measured the interaction forces upon approach and retraction of thus made tips. Waterborne Raoultella terrigena and Escherichia coli adhered weakly (1-2 nN) to different activated carbon particles, and the main difference between the activated carbons was the percentage of curves with attractive sites revealed upon traversing of a carbon particle through the bacterial EPS layer. The percentage of curves showing adhesion forces upon retraction varied between 21% and 69%, and was highest for R. terrigena with positively charged carbon (66%) and a coconut carbon (69%). Macroscopic bacterial removal by the mesoporous carbon particles increased with increasing percentages of attractive sites revealed upon traversing a carbon particle through the outer bacterial surface layer. PMID- 18405911 TI - The terminal rise velocity of 10-100 microm diameter bubbles in water. AB - Single bubbles of very pure N2, He, air and CO2 were formed in a quiescent environment in ultra-clean water, with diameters ranging from 10 to 100 mum. Their terminal rise velocities were measured by high-speed video microscopy. For N2, He and air, excellent agreement with the Hadamard-Rybczynski (H-R) equation was observed, indicating that slip was occurring at the liquid-vapor interface. For CO2 bubbles with diameters less than 60 microm, the terminal rise velocities exceeded those predicted by the H-R equation. This effect was ascribed to the enhanced solubility of CO2 compared with the other gases examined. The presence of a diffusion boundary layer may be responsible for the increased terminal velocity of very small CO2 bubbles. PMID- 18405912 TI - Effect of magnesium/calcium ratios in solutions treated by electrodialysis: morphological characterization and identification of anion-exchange membrane fouling. AB - The present study aimed the characterization of the fouling formed on anion exchange membrane during electrodialysis treatment of model salt solutions at different Mg/Ca ratio (0, 1/20, 1/10, 1/5 and 2/5). The membrane fouling was characterized by membrane parameters (membrane thickness and electrical conductivity) and identified by membrane surface analysis (elemental analysis and X-ray diffraction). The mineral deposit was identified as Ca(OH)2 when no magnesium was present in the model salt. As soon as magnesium was present in the model salt solution for neutral pH((concentrate)) conditions a mix between CaCO3 and Ca(OH)2 was formed. This study is the first one to report the influence of magnesium in solution on the formation of CaCO3 fouling at the interface of anion exchange membrane during electrodialysis. PMID- 18405913 TI - Mechanism for stabilization of the molten globule state of papain by sodium n alkyl sulfates: spectroscopic and calorimetric approaches. AB - Papain exists in a molten globule (MG) state at pH 2 and in this state protein tends to aggregate in the presence of lower concentrations of guanidine hydrochloride (GuHCl). Such aggregation is prevented if low concentrations of sodium n-alkyl sulfates are also present in the buffer; in addition, stabilization of the protein is also induced. The guanidine hydrochloride and temperature-induced unfolding of papain, in the presence of n-alkyl sulfates, indicate stabilization of the protein as seen from the higher transition midpoints when monitored by fluorescence, circular dichroism, and differential scanning calorimetry. However, a similar phenomenon is not seen under neutral conditions in the presence of n-alkyl sulfate concentrations. The effect of n alkyl sulfates on the structure of the MG state of papain was utilized to investigate the contribution of hydrophobic interaction to the stability of the MG state. The Td values of the MG states of papain in the presence of n-alkyl sulfates at different concentrations showed substantial variation. The enhancement of Td values at the stability criterion of MG states corresponded with increasing chain length of the cited n-alkyl sulfates. The present results suggest that the hydrophobic interactions play important roles in stabilizing and preventing the aggregation of the MG state of papain. PMID- 18405914 TI - Bite-block perturbation in people who stutter: immediate compensatory and delayed adaptive processes. AB - This exploratory study investigated sensory-motor mechanisms in five people who stutter (PWS) and five people who do not (PNS). Lip kinematic and coordination data were recorded as they produced bi-syllabic nonwords at two rates (normal and fast) in three conditions (jaw-free, immediately after insertion of a bite-block, and after a 10-min accommodation period). At normal speech rates, effects of bite blocks on lip kinematics were similar for both PWS and PNS speakers showing larger amplitudes, peak velocities, shorter durations and more stable movement cycle patterns. However, at fast speech rates upper lip responses of PWS exhibited larger amplitudes and peak velocities. At both speech rates, the presence of a bite-block changed movement coordination patterns only for PNS. However, at fast speech rates bite-blocks decreased variability of coordination patterns for both groups. No adaptive changes in movement stability were found for either group, but a practice-related increase in lower lip peak velocity was found at normal speech rates. These findings indicate that bite-block perturbation did not exacerbate any hypothesized limitation or difficulty in controlling individual articulatory movements or their coordination in PWS. The results also support the position that specific motor control strategies are used by PWS as compared to PNS to compensate for bite-block perturbations under increased speech rate demands. LEARNING OUTCOMES: The reader will be able to: (1) distinguish between compensatory and adaptive responses to bite-block perturbation; (2) explain the measurement of articulatory stability; (3) summarize the potential role of motor control strategies in people who stutter; and (4) discuss the assumptions of the motor skills approach to stuttering. PMID- 18405915 TI - Genetic basis of sex pheromone blend difference between Helicoverpa armigera (Hubner) and Helicoverpa assulta (Guenee) (Lepidoptera: Noctuidae). AB - The two closely related moth species, Helicoverpa armigera and H. assulta, are sympatric in China. Both species use a mixture of (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-9-hexadecenal (Z9-16:Ald) as their sex pheromones but in widely different ratios. Hybridization and backcrossing experiments between H. armigera and H. assulta were conducted and sex pheromone compositions of the parent species, their F(1) hybrids and backcrosses were compared to study the genetic basis of the production of their sex pheromone blend composition. Results show that the difference in sex pheromone blend ratios of these Helicoverpa species is mainly controlled by an autosomal locus with two alleles, with the allele from H. armigera being almost completely dominant over that derived from H. assulta. PMID- 18405916 TI - Involvement of connexin 43 in angiotensin II-induced migration and proliferation of saphenous vein smooth muscle cells via the MAPK-AP-1 signaling pathway. AB - Proliferation and migration of vascular smooth muscle cells (VSMCs) lead to intimal thickening and influence the long-term patency of venous graft post coronary arterial bypass graft. There is increasing evidence that connexins are involved in the development of intimal hyperplasia and restenosis. We assessed connexin 43 (Cx43) expression and its role in angiotensin II-induced proliferation and migration of smooth muscle cells and the signal pathways involved in human saphenous vein bypass conduits. Angiotensin II significantly increased gap junctional intercellular communication and induced the expression of Cx43 in human saphenous vein SMCs in a dose- and time-dependent manner through angiotensin II type 1 receptor. The effect of angiotensin II was blocked by siRNA of ERK 1/2, p38 MAPK and JNK, respectively. Overexpression of Cx43 markedly increased the proliferation of saphenous vein SMCs. However, siRNA for Cx43 inhibited angiotensin II-induced proliferation, cyclin E expression and migration of human saphenous vein SMCs. In dual-luciferase reporter assay, angiotensin II markedly activated AP-1 transcription factor, which was significantly attenuated by a dominant-negative AP-1 (A-Fos) with subsequent inhibition of angiotensin II induced transcriptional expression of Cx43. These data demonstrate the role of Cx43 in the proliferation and migration of human saphenous vein SMCs and angiotensin II-induced Cx43 expression via mitogen-activated protein kinases (MAPK)-AP-1 signaling pathway. PMID- 18405917 TI - Integrin alpha IIb beta 3 in a membrane environment remains the same height after Mn2+ activation when observed by cryoelectron tomography. AB - Integrins perform the critical function of signalling cell attachment to the extracellular matrix or to other cells. This signalling is done through a structural change propagated bidirectionally across the plasma membrane. Integrin activation has been extensively studied with ectodomain constructs, but the structural change within intact, membrane-bound molecules remains a subject of live debate. Using cryoelectron tomography, we examined the simplest predication of the different integrin activation models, i.e., the change in height of the molecules. Analysis using techniques that compensate for the missing wedge during alignment and averaging and that search for patterns in the structure of the aligned molecular subvolumes extracted from the tomogram reveals that the vast majority of molecules show no dramatic height change upon Mn(2+)-induced activation of membrane-bound integrins when compared with an inactive integrin control group. Thus, the result is inconsistent with the switchblade activation model. PMID- 18405919 TI - Diminished prefrontal oxygenation with normal and above-average verbal fluency performance in adult ADHD. AB - Attention-Deficit/Hyperactivity Disorder (ADHD) in adults is associated with deficits in executive functions such as verbal fluency. Functional near-infrared spectroscopy offers the possibility to measure brain activity during verbal fluency in psychiatric patients due to low susceptibility to movements artefacts. Fourteen patients with mainly combined ADHD subtype and 14 healthy controls matched for age, gender, handedness, education level, and intelligence showed equal performance in phonological verbal fluency and higher performance in semantical verbal fluency in comparison to the controls. On the level of brain function indicated by concentration changes of oxygenated and deoxygenated haemoglobin, both groups showed inferior frontal brain activity during both tasks. ADHD patients had a lower magnitude of oxygenation and a significant negative correlation of brain activity with performance, i.e. higher brain activity was associated with lower performance. These results might be interpreted as a lack of disease related deficits, as an economical recruitment of cognitive resources, or - more likely - as an expression of a benefit in the patient group. High verbal competence in our samples could contribute to these findings. One speculative and post hoc explanation aims at the clinically well known phenomenon of hyperfocusing in patients with ADHD. PMID- 18405918 TI - A unique mode of microtubule stabilization induced by peloruside A. AB - Microtubules are significant therapeutic targets for the treatment of cancer, where suppression of microtubule dynamicity by drugs such as paclitaxel forms the basis of clinical efficacy. Peloruside A, a macrolide isolated from New Zealand marine sponge Mycale hentscheli, is a microtubule-stabilizing agent that synergizes with taxoid drugs through a unique site and is an attractive lead compound in the development of combination therapies. We report here unique allosteric properties of microtubule stabilization via peloruside A and present a structural model of the peloruside-binding site. Using a strategy involving comparative hydrogen-deuterium exchange mass spectrometry of different microtubule-stabilizing agents, we suggest that taxoid-site ligands epothilone A and docetaxel stabilize microtubules primarily through improved longitudinal interactions centered on the interdimer interface, with no observable contributions from lateral interactions between protofilaments. The mode by which peloruside A achieves microtubule stabilization also involves the interdimer interface, but includes contributions from the alpha/beta-tubulin intradimer interface and protofilament contacts, both in the form of destabilizations. Using data-directed molecular docking simulations, we propose that peloruside A binds within a pocket on the exterior of beta-tubulin at a previously unknown ligand site, rather than on alpha-tubulin as suggested in earlier studies. PMID- 18405920 TI - Modeling local and global intracellular calcium responses mediated by diffusely distributed inositol 1,4,5-trisphosphate receptors. AB - Considerable insight into intracellular Ca2+ responses has been obtained through the development of whole cell models that are based on molecular mechanisms, e.g., single channel kinetics of the inositol 1,4,5-trisphosphate (IP3) receptor Ca2+ channel. However, a limitation of most whole cell models to date is the assumption that IP3 receptor Ca2+ channels (IP3Rs) are globally coupled by a "continuously stirred" bulk cytosolic [Ca2+], when in fact open IP3Rs experience elevated "domain" Ca2+ concentrations. Here we present a 2N+2-compartment whole cell model of local and global Ca2+ responses mediated by N=100,000 diffusely distributed IP3Rs, each represented by a four-state Markov chain. Two of these compartments correspond to bulk cytosolic and luminal Ca2+ concentrations, and the remaining 2N compartments represent time-dependent cytosolic and luminal Ca2+ domains associated with each IP3R. Using this Monte Carlo model as a starting point, we present an alternative formulation that solves a system of advection reaction equations for the probability density of cytosolic and luminal domain [Ca2+] jointly distributed with IP3R state. When these equations are coupled to ordinary differential equations for the bulk cytosolic and luminal [Ca2+], a realistic but minimal model of whole cell Ca2+ dynamics is produced that accounts for the influence of local Ca2+ signaling on channel gating and global Ca2+ responses. The probability density approach is benchmarked and validated by comparison to Monte Carlo simulations, and the two methods are shown to agree when the number of Ca2+ channels is large (i.e., physiologically realistic). Using the probability density approach, we show that the time scale of Ca2+ domain formation and collapse (both cytosolic and luminal) may influence global Ca2+ oscillations, and we derive two reduced models of global Ca2+ dynamics that account for the influence of local Ca2+ signaling on global Ca2+ dynamics when there is a separation of time scales between the stochastic gating of IP3Rs and the dynamics of domain Ca2+. PMID- 18405921 TI - Are the local adjustments of the relative spatial frequencies of the dynein arms and the beta-tubulin monomers involved in the regulation of the "9+2" axoneme? AB - The "9+2" axoneme is a highly specific cylindrical machine whose periodic bending is due to the cumulative shear of its 9 outer doublets of microtubules. Because of the discrete architecture of the tubulin monomers and the active appendices that the outer doublets carry (dynein arms, nexin links and radial spokes), this movement corresponds to the relative shear of these topological verniers, whose characteristics depend on the geometry of the wave train. When an axonemal segment bends, this induces the compressed and dilated conformations of the tubulin monomers and, consequently, the modification of the spatial frequencies of the appendages that the outer doublets carry. From a dynamic point of view, the adjustments of the spatial frequencies of the elements of the two facing verniers that must interact create different longitudinal periodic patterns of distribution of the joint probability of the molecular interaction as a function of the location of the doublet pairs around the axonemal cylinder and their spatial orientation within the axonemal cylinder. During the shear, these patterns move along the outer doublet intervals at a speed that ranges from one to more than a thousand times that of sliding, in two opposite directions along the two opposite halves of the axoneme separated by the bending plane, respecting the polarity of the dynein arms within the axoneme. Consequently, these waves might be involved in the regulation of the alternating activity of the dynein arms along the flagellum, because they induce the necessary intermolecular dialog along the axoneme since they could be an element of the local dynamic stability/instability equilibrium of the axoneme. This complements the geometric clutch model [Lindemann, C., 1994. A "geometric clutch" hypothesis to explain oscillations of the axoneme of cilia and flagella. J. Theor. Biol. 168, 175-189]. PMID- 18405922 TI - Evolutionary stability conditions for signaling games with costly signals. AB - The paper investigates the class of signaling games with the following properties: (a) the interests of sender and receiver coincide, (b) different signals incur differential costs, and (c) different events (meanings/types) have different probabilities. Necessary and sufficient conditions are presented for a profile to be evolutionarily stable and neutrally stable, and for a set of profiles to be an evolutionarily stable set. The main finding is that a profile belongs to some evolutionarily stable set if and only if a maximal number of events can be reliably communicated. Furthermore, it is shown that under the replicator dynamics, a set of states with a positive measure is attracted to "sub optimal" equilibria that do not belong to any asymptotically stable set. PMID- 18405923 TI - Capsaicin induces apoptosis and terminal differentiation in human glioma A172 cells. AB - Capsaicin (8-methyl-N-vanillyl-6-nonenamide), the major pungent ingredient of red pepper, has been reported to possess anti-carcinogenic and anti-mutagenic activities. In this study, the effects of capsaicin on human glioblastoma A172 cells were investigated. Treatment of A172 cells with capsaicin inhibited cell growth and induced apoptosis through down-regulation of Bcl-2 and activation of caspase-3. Interestingly, synergistic induction of morphological alternation was observed when A172 cells were treated with capsaicin. A double immunostaining analysis indicated that capsaicin stimulated terminal differentiation predominantly to astrocyte-like cells. Moreover, capsaicin increased the transcription levels of glial fibrillary acidic protein (GFAP) and neuronal microtubule-associated protein 2ab (MAP2ab). These results demonstrated that capsaicin inhibits A172 cell growth through apoptosis and terminal differentiation. Consequently, this research may provide further support for capsaicin-based anti-tumor therapies and consideration should be given to developing capsaicin for use in chemotherapy for malignant human glioblastoma. PMID- 18405924 TI - Contamination of marine biogenic habitats and effects upon associated epifauna. AB - Habitat-forming organisms are frequently used as biomonitors in marine environments due to a widespread ability to accumulate toxic contaminants. Few studies, however, have considered the consequences of these accumulated contaminants on the abundant and diverse fauna associated with these habitats. In this review, we summarize research which has investigated the contamination of biogenic habitats (including seagrasses, macroalgae, ascidians, sponges and bivalve reefs) and the impact of this contamination on the habitat use, feeding behaviour and survival of associated epifauna. In many cases, ecological impacts upon epifauna are not simply predicted by levels of contamination in their habitat, but are determined by the foraging, feeding and reproductive behaviours of the inhabiting organisms. Thus, a thorough understanding of these ecological processes is essential in order to understand the effects of contaminants upon epifaunal communities. The scope of biomonitoring studies which assess the contamination of biogenic habitats should be expanded to include an assessment of potential effects upon associated epifauna. When combined with manipulative field experiments such an approach would greatly assist in our understanding of indirect effects of contaminants in these important benthic habitats. PMID- 18405925 TI - Relationship between macrobenthos and physical habitat characters in tidal flat in eastern Seto Inland Sea, Japan. AB - The investigations were carried out at 6 tidal flats located on the eastern part of the Seto Inland Sea, Japan. This study was focused on physical characteristics of sediments, namely as particle size of sediment and difference in elevation, and generalizes the relationship between sediments and macrobenthos. A total of 192 species were collected at 187 stations at 6 tidal flats. Physical characteristics of sediment were classified into 9 groups by cluster analysis in relation to sediment particle size and difference in elevation. Those groups had also significant difference in physical characteristics of sediments, and were characterized by some specific macrobenthos species. Distribution of macrobenthos can be explained by the classification of physical characteristics of sediment. These findings show the possibility to predict the variety of macrobenthos community using the physical characteristics of sediment. PMID- 18405926 TI - Computation of threshold conditions for epidemiological models and global stability of the disease-free equilibrium (DFE). AB - One goal of this paper is to give an algorithm for computing a threshold condition for epidemiological systems arising from compartmental deterministic modeling. We calculate a threshold condition T(0) of the parameters of the system such that if T(0)<1 the disease-free equilibrium (DFE) is locally asymptotically stable (LAS), and if T(0)>1, the DFE is unstable. The second objective, by adding some reasonable assumptions, is to give, depending on the model, necessary and sufficient conditions for global asymptotic stability (GAS) of the DFE. In many cases, we can prove that a necessary and sufficient condition for the global asymptotic stability of the DFE is R(0)< or =1, where R(0) is the basic reproduction number [O. Diekmann, J.A. Heesterbeek, Mathematical Epidemiology of Infectious Diseases: Model Building, Analysis and Interpretation, Wiley, New York, 2000]. To illustrate our results, we apply our techniques to examples taken from the literature. In these examples we improve the results already obtained for the GAS of the DFE. We show that our algorithm is relevant for high dimensional epidemiological models. PMID- 18405927 TI - Molecular characterization and phylogenetic analysis of new Newcastle disease virus isolates from the mainland of China. AB - Seventy-nine velogenic Newcastle disease virus (NDV) isolates were obtained from infected chicken flocks during the outbreaks of Newcastle disease (ND) in various regions of the mainland of China in 2006. The F gene fragment (535bp, from nt 47 to 581 of the F gene) which codes the main functional region of the F protein was obtained by RT-PCR and sequenced. All sequences obtained in this study have been submitted to GenBank. All the isolates have the motif (112)R-R-Q/R-K/R-R-F(117) at the cleavage site of the fusion protein, which is typical of velogenic NDV isolates. For genotyping, a phylogenetic tree based on nucleotides 47-435 of the F gene was constructed, and the 79 isolates could be divided into two genotypes, namely VIId and III. Most of the isolates proved to be of genotype VIId; only two isolates were of genotype III. Genotype VIId NDV has been the predominant pathogen responsible for most Newcastle disease outbreaks in China. The proportion of isolates of genotype VIId NDV shows an increasing trend, according to studies on the molecular epidemiology of NDV in China from 2002 to 2006. PMID- 18405928 TI - The future looks bright: low-cost fluorescent microscopes for detection of Mycobacterium tuberculosis and Coccidiae. AB - In many resource-poor countries, diagnosis of tuberculosis is based on microscopy of Ziehl-Neelsen-stained smears. However, the fluorescent stain auramine performs better and allows the detection of more positive smears. The limitation for its widespread use has been the high cost for fluorescent microscopes, especially the frequent replacement of the expensive light source, which lasts only 200-300 h. Novel light-emitting diodes (LED) are alternatives and last ten thousands of hours. Based on LEDs, rather inexpensive solutions for fluorescent microscopes are now available and thus auramine may be a cost-effective step to improve the diagnosis of tuberculosis in resource-poor countries. PMID- 18405929 TI - Anaemia and malaria in Yanomami communities with differing access to healthcare. AB - Inequitable access to healthcare has a profound impact on the health of marginalised groups that typically suffer an excess burden of infectious disease morbidity and mortality. The Yanomami are traditionally semi-nomadic people living in widely dispersed communities in Amazonian Venezuela and Brazil. Only communities living in the vicinity of a health post have relatively constant access to healthcare. To monitor the improvement in the development of Yanomami healthcare a cross-sectional survey of 183 individuals was conducted to investigate malaria and anaemia prevalence in communities with constant and intermittent access to healthcare. Demographic and clinical data were collected. Malaria was diagnosed by microscopy and haemoglobin concentration by HemoCue. Prevalence of malaria, anaemia, splenomegaly, fever and diarrhoea were all significantly higher in communities with intermittent access to healthcare (anaemia 80.8% vs. 53.6%, P<0.001; malaria 18.2% vs. 6.0%, P=0.013; splenomegaly 85.4% vs.12.5%, P<0.001; fever 50.5% vs. 28.6%, P=0.003; diarrhoea 30.3% vs.10.7% P=0.001). Haemoglobin level (10.0 g/dl vs. 11.5 g/dl) was significantly associated with access to healthcare when controlling for age, sex, malaria and splenomegaly (P=0.01). These findings indicate a heavy burden of anaemia in both areas and the need for interventions against anaemia and malaria, along with more frequent medical visits to remote areas. PMID- 18405930 TI - Characterisation of DDT, pyrethroid and carbamate resistance in Anopheles funestus from Obuasi, Ghana. AB - Indoor-resting anopheline mosquitoes were collected from Obuasi, Ghana, and were identified morphologically and by PCR as Anopheles funestus Giles. Wild-caught females were induced to lay eggs. Samples of F1 progeny from each family were divided into cohorts and were either exposed to DDT and permethrin or were stored for biochemical analysis. Bioassay data by family show evidence of DDT and pyrethroid resistance in the parent A. funestus population. The sodium channel gene of DDT survivors and DDT-susceptible individuals was PCR amplified and sequenced to determine whether any kdr-type mutations were present. Molecular analysis of the IIS5-IIS6 segment of the sodium channel gene gave no indication of any kdr-type mutations associated with resistance phenotypes. Biochemical analysis suggests that DDT and pyrethroid resistance may be metabolically mediated, although there were no clear correlations between enzyme levels/activities and insecticide resistance across families. Furthermore, an altered acetylcholinesterase conferring carbamate resistance was evident. These results can be used to plan an effective malaria control strategy in the Obuasi region. PMID- 18405931 TI - 17beta-estradiol attenuates cardiac dysfunction and decreases NF-kappaB binding activity in mechanically stretched rat hearts. AB - This study was to examine the effect of estrogen on mechanical stretching-induced cardiac dysfunction in an isolated heart model. The isolated rat hearts were perfused via the Langendorff system and exposed to left ventricular stretching. One group hearts (n=6) were perfused with 17beta-estradiol (100nM) and the other group hearts (n=6) were perfused with estrogen plus its receptor antagonist ICI182,780 (1microM) before myocardial stretching was performed. Control hearts (n=6) were perfused with perfusion buffer. Cardiac functions were recorded. At the end of perfusion, the hearts were harvested and the levels of tumor necrosis factor-alpha (TNF-alpha), phospho-p38 mitogen-activated protein kinase (MAPK) and nuclear factor-kappaB (NF-kappaB) binding activity were examined. Acute ventricular stretching resulted in significantly decrease in left ventricular developed pressure (LVDP) by 42.7%, maximal positive and negative values of the first derivative of pressure (+dP/dt and -dP/dt) by 43.2%, and 43.5%, respectively. The levels of TNF-alpha, phospho-p38 MAPK and NF-kappaB DNA binding activity were significantly increased following myocardial stretching. In 17beta estradiol treated hearts, the myocardial functions were significantly improved. The levels of TNF-alpha, phospho-p38 MAPK, and NF-kappaB binding activity in myocardium were also significantly reduced by 35.7%, 56.9%, and 50%, respectively, compared with untreated stretched hearts. The beneficial effects of 17beta-estradiol on the stretched hearts were abolished by ICI182,780. The results suggest that pharmacological dose of 17beta-estradiol will attenuate stretching-induced cardiac dysfunction in an isolated heart model. The mechanisms could involve in blunting p38 MAPK and NF-kappaB signaling. PMID- 18405932 TI - Morphological changes and EGF expression in the granular convoluted tubule cells of submandibular glands of Trypanosoma cruzi infected rats. AB - We have previously demonstrated in rats that Chagas' disease affects the salivary glands, by promoting an enlargement of the submandibular gland. In order to further investigate possible functional alterations on infected submandibular glands, the objective of the present study was to analyze epidermal growth factor (EGF) expression on rat submandibular glands during Trypanosoma cruzi infection. Results demonstrated that infected rats presented lower levels of testosterone, and morphological changes in the granular convoluted tubule (GCT) cells of the submandibular glands, along with acinar enlargement and delayed ductal maturation at the developing granular ducts. Immunohistochemistry analysis additionally showed that only few cells immunolabelled with anti-EGF on infected rats during the acute phase of Chagas' disease, while after 64 and 90 days (chronic phase) of infection, EGF expression was similar to non-infected rats. The present findings suggest that at the acute phase of Chagas' disease, lower levels of testosterone may lead to a delayed maturation of GCT, which positively correlates with decreased EGF production by submandibular glands cells. PMID- 18405933 TI - Analysis of chemokines and reactive oxygen species formation by rat and human neutrophils induced by microcystin-LA, -YR and -LR. AB - Microcystins (MC), a family of heptapeptide toxins produced by some genera of Cyanobacteria, have potent hepatotoxicity and tumor-promoting activity. Leukocyte infiltration in the liver was observed in MC-induced acute intoxication. Although the mechanisms of hepatotoxicity are still unclear, neutrophil infiltration in the liver may play an important role in triggering toxic injury and tumor development. The present study reports the effects of MC-LA, MC-YR and MC-LR (1 and 1000 nM) on human and rat neutrophils functions in vitro. Cell viability, DNA fragmentation, mitochondrial membrane depolarization and intracellular reactive oxygen species (ROS) levels were measured by flow cytometry. Extracellular ROS content was measured by lucigenin-amplified chemiluminescence, and cytokines were determined by ELISA. We found that these MC increased interleukin-8 (IL-8), cytokine-induced neutrophil chemoattractant-2alphabeta (CINC-2alphabeta) and extracellular ROS levels in human and rat neutrophils. Apart from neutrophil presence during the inflammatory process of MC-induced injury, our results suggest that hepatic neutrophil accumulation is further increased by MC-induced neutrophil-derived chemokine. PMID- 18405934 TI - Cloning, characterization and phylogenetic analyses of members of three major venom families from a single specimen of Walterinnesia aegyptia. AB - Walterinnesia aegyptia is a monotypic elapid snake inhabiting in Africa and Mideast. Although its envenoming is known to cause rapid deaths and paralysis, structural data of its venom proteins are rather limited. Using gel filtration and reverse-phase HPLC, phospholipases A(2) (PLAs), three-fingered toxins (3FTxs), and Kunitz-type protease inhibitors (KIns) were purified from the venom of a single specimen of this species caught in northern Egypt. In addition, specific primers were designed and PCR was carried out to amplify the cDNAs encoding members of the three venom families, respectively, using total cDNA prepared from its venom glands. Complete amino acid sequences of two acidic PLAs, three short chain 3FTxs, and four KIns of this venom species were thus deduced after their cDNAs were cloned and sequenced. They are all novel sequences and match the mass data of purified proteins. For members of each toxin family, protein sequences were aligned and subjected to molecular phylogenetic analyses. The results indicated that the PLAs and a Kunitz inhibitor of W. aegyptia are most similar to those of king cobra venom, and its 3FTxs belongs to either Type I alpha-neurotoxins or weak toxins of orphan-II subtype. It is remarkable that both king cobra and W. aegyptia cause rapid deaths of the victims, and a close evolutionary relationship between them is speculated. PMID- 18405935 TI - Using a filtering task to measure the spatial extent of selective attention. AB - The spatial extent of attention was investigated by measuring sensitivity to stimuli at to-be-ignored locations. Observers detected a stimulus at a cued location (target), while ignoring otherwise identical stimuli at nearby locations (foils). Only an attentional cue distinguished target from foil. Several experiments varied the contrast and separation of targets and foils. Two theories of selection were compared: contrast gain and a version of attention switching called an all-or-none mixture model. Results included large effects of separation, rejection of the contrast gain model, and the measurement of the size and profile of the spatial extent of attention. PMID- 18405936 TI - 17alpha-Ethinylestradiol sorption to activated sludge biomass: thermodynamic properties and reaction mechanisms. AB - Sorption is an important mechanism for removal of 17alpha-ethinylestradiol (EE(2)) in biological wastewater treatment. In an effort to deepen our understanding of this process, this work investigated the underlying thermodynamic parameters. Biomass was harvested from a membrane bioreactor (MBR) and a sequencing batch reactor (SBR), and sorption experiments were conducted over a range of temperatures. Sorption of EE(2) to activated sludge was spontaneous (DeltaG values were between -16 and -11 KJ/mol), enthalpy-driven (DeltaH values were -37 KJ/mol (MBR) and -48 KJ/mol (SBR)), and entropy-retarded (DeltaS values were -74 (MBR) and -119 J/mol/K (SBR)). Although EE(2) is nonpolar, hydrophobic interactions were not dominant driving forces. The thermodynamic data also suggested that EE(2) sorption to biomass was primarily physisorption, but it also included low-level chemisorption. The FT-IR results suggested that chemical reactions were not significant enough to shift the detectable chemical bonding characteristics of the biomass functional groups. PMID- 18405937 TI - Modeling the competitive effect of ammonium oxidizers and heterotrophs on the degradation of MTBE in a packed bed reactor. AB - A mathematical model was used to study effects on the degradation of methyl tert butyl ether (MTBE) in a packed bed reactor due to the presence of contaminants such as ammonium, and the mix of benzene, toluene, ethylbenzene and xylenes (BTEX). It was shown that competition between the slower growing MTBE degraders and the co-contaminant oxidizers prevented MTBE's degradation when oxygen was limited. In this event, the co-contaminant oxidizers out-competed the MTBE degraders in the reactor's biofilm. However, if the oxygen supply was sufficient, MTBE would be fully degraded after the zone where the co-contaminants were oxidized. The results of the model further indicate that contradicting findings in the literature about the effects of BTEX on the degradation of MTBE are mainly due to differences in the study methodologies. Effects such as short-term toxicity of BTEX and the lack of steady-state conditions may also add to contradictions among reports. PMID- 18405938 TI - Biofilm processes in biologically active carbon water purification. AB - This review paper serves to describe the composition and activity of a biologically active carbon (BAC) biofilm used in water purification. An analysis of several physical-chemical, biochemical and microbiological methods (indicators) used to characterize the BAC biofilm's composition and activity is provided. As well, the ability of the biofilm to remove and biodegrade waterborne organic substances and pollutants will be reviewed, with context to other industrial processes such as pre-ozonation and post-membrane filtration. Strategies to control the growth of the BAC biofilm, such as varying the nutrient loading rate, manipulating influent DO and pH levels, altering the frequency of BAC filter backwashing and applying oxidative disinfection, will be described in detail along with their respective process control challenges. PMID- 18405939 TI - Ozonation catalyzed by homogeneous and heterogeneous catalysts for degradation of DEHP in aqueous phase. AB - Di-(2-ethyl hexyl) phthalate (DEHP), a recalcitrant and an endocrine disrupting chemical, was studied for its removal from wastewater by advanced oxidation process. The effects of pH, transition metal ions, and granular activated carbon (GAC) were investigated. Removal of DEHP increased with increase in pH and among metal ions Cr(III) was found to be the most active catalyst to remove DEHP. In the case of GAC, original carbon (G0) and GAC pre-ozonated in gas phase for 10 min (G10) were tested as catalysts in catalytic ozonation and found G0 to be more active than G10. This is because, during pre-ozonation, basic groups like chromene, pyrones and also graphene layers were oxidized to acidic surface oxygen groups. These basic surface groups are reported to be active catalytic centres for ozone decomposition into .OH which is a strong oxidant. According to kinetic manipulations, DEHP degradation rate constant due to .OH was affected by the catalyst while that due to direct ozonation is same in all cases with or without catalyst. G0 was doped with chromia gels (G0/CrA) to combine the benefits of homogeneous and heterogeneous catalysis. G0/CrA showed lower catalytic efficiency than that of only G0. This might be because of changes in surface structure of GAC caused by doping of chromia gel and changes in chemical nature of Cr(III) during formation of gel. A good correlation was found between the rate constants of ozone decay and DEHP degradation (R2=0.96). This correlation confirms that ozone decomposition into .OH is a critical factor for the activity of a catalyst during ozonation. PMID- 18405940 TI - Variation in sensitivity to cadmium among genetically characterized laboratory strains of the midge Chironomus riparius. AB - Estimating the extent of intraspecific variation in sensitivity to contaminant exposure is important in order to explain variation in the outcome of toxicity tests and to predict the effects of chemical stress on natural populations of plants and animals. However, only few studies provide evident data concerning intraspecific variation in life-history traits caused by a differential response to chemical stress. In this study, we compared the life-history response of six laboratory strains of the midge Chironomus riparius to cadmium exposure in a full life-cycle assay. In addition, the level of genetic variation in all strains was measured at five variable microsatellite loci. Several significant differences in life-history traits among the strains were observed in controls and cadmium treatments. The extent of variation among strains was largest at moderate cadmium concentration (0.42 mg Cd/kg dw). At increased Cd concentrations all strains showed similar levels of high mortality and reduced reproductive success. All strains exhibited considerable levels of genetic impoverishment compared to field populations. Strains with low genetic variation showed reduced fitness in the controls and were more susceptible to Cd exposure. For instance, no reproductive success in the lowest Cd treatment was observed for the strain with the lowest level of genetic diversity. In contrast, this Cd concentration had no negative effects on life-history traits of more variable strains. These results confirm recent findings, that inbreeding and reduced genetic variation influence the reaction of populations towards environmental stress. In addition, we show that the level of genetic variation and inbreeding directly influences the outcome of toxicity tests and contributes to the lack of reproducibility of test results among laboratories. PMID- 18405941 TI - Coupled solar photo-Fenton and biological treatment for the degradation of diuron and linuron herbicides at pilot scale. AB - A coupled solar photo-Fenton (chemical) and biological treatment has been used to remove biorecalcitrant diuron (42 mg l(-1)) and linuron (75 mg l(-1)) herbicides from water at pilot plant scale. The chemical process has been carried out in a 82 l solar pilot plant made up by four compound parabolic collector units, and it was followed by a biological treatment performed in a 40 l sequencing batch reactor. Two Fe(II) doses (2 and 5 mg l(-1)) and sequential additions of H2O2 (20 mg l(-1)) have been used to chemically degrade the initially polluted effluent. Next, biodegradability at different oxidation states has been assessed by means of BOD/COD ratio. A reagent dose of Fe=5 mg l(-1) and H2O2=100 mg l(-1) has been required to obtain a biodegradable effluent after 100 min of irradiation time. Finally, the organic content of the photo-treated solution has been completely assimilated by a biomass consortium in the sequencing batch reactor using a total suspended solids concentration of 0.2 g l(-1) and a hydraulic retention time of 24h. Comparison between the data obtained at pilot plant scale (specially the one corresponding to the chemical step) and previously published data from a similar system performing at laboratory scale, has been carried out. PMID- 18405942 TI - Drinking water intake by infants living in rural Quebec (Canada). AB - Drinking water is a potential source of toxic contaminant and it is well known that water intake on a body weight basis decreases rapidly with increasing age. Nevertheless, few studies have been conducted on water intake of very young infants who might be particularly sensitive to some toxic chemicals. The objective of this study was to describe the mean and distribution of total water intake of 2-month old infants living in agricultural areas. Mothers (n=642) of 8 to 9 week old infants were interviewed by phone to evaluate their feeding practice, including juice and cereal intake. There were 393 infants (61%) who drank some quantity of water and 278 (43%) consumed formula reconstituted with water. For formula-fed infants, the 10, 50 and 90th percentiles of daily water intake were 79, 112, and 179 ml/kg respectively. These values are much higher than the intake recommended by US EPA for infants under one year (US EPA, 1997). This study demonstrates the importance of considering water distribution intake in very young infants who may be particularly susceptible to water contaminants. PMID- 18405943 TI - Development of an inventory and temporal allocation profiles of emissions from power plants and industrial facilities in Thailand. AB - An emission inventory (EI) of power plants and industrial (i.e., non-power plant) facilities in Thailand was developed. Emissions considered are those from fuel consumption (i.e., combustion) for power plants and those from both fuel consumption and industrial processes (i.e., non-combustion) for industrial facilities. For power plants, total annual emissions due to fuel consumption are 107.9 x 10(3) ton NOx (as NO2), 146.2 x 10(3) ton SO2, 6.1 x 10(3) ton NMVOC (non methane volatile organic compounds), 47.0 x 10(3) ton CO, 1.8 x 10(3) ton NH3, 1.5 x 10(3) ton OC (organic carbon), and 1.5 x 10(3) ton BC (black carbon). For industrial facilities, total annual emissions due to fuel consumption are 111.4 x 10(3) ton NOx (as NO2), 476.9 x 10(3) ton SO2, 33.4 x 10(3) ton NMVOC, 193.1 x 10(3) ton CO, 1.6 x 10(3) ton NH3, 8.5 x 10(3) ton OC, and 8.0 x 10(3) ton BC. Among various industrial types, Food and Beverage, Chemical, and Non-Metal industries are dominant emitters. Total annual emissions due to industrial processes are 79.2 x 10(3) ton SO2, 76.0 x 10(3) ton NMVOC, and 4.8 x 10(3) ton CO. The Central and Eastern regions combined contribute considerably to total emissions for most emission species. Emission estimates found here show fair agreement with those in some selected past studies. A crude estimation of potential fugitive NMVOC emissions specifically from petroleum industry was also made, and the estimates found could be considered significant (nearly half of NMVOC emissions from industrial processes). Several temporal allocation profiles of emissions were also developed and suggested for power plants and industrial facilities, including monthly, daily, and hourly profiles. PMID- 18405944 TI - A possible role of insulin-like growth factor-II C-peptide in regulating the function of steroidogenic cells in adult frog adrenal glands. AB - The sole structural determinant for the differential ability of the insulin-like growth factors (IGF-I and IGF-II) to induce autophosphorylation of specific insulin receptor (IR) tyrosine residues and activate downstream signaling molecules is the C domain. The IR is structurally related to the type I insulin like growth factor receptor (IGF-IR). This study aimed to identify the presence of IGF receptors by which the IGF-II C-peptide could mediate its effects in the frog (Rana ridibunda) adrenal glands and to observe whether injection of IGF-II C peptide affects the function of adrenal steroidogenic cells using light and transmission electron microscopy and by the evaluation of the immunoreactivity of steroidogenic acute regulatory protein (StAR). After IGF-II C-peptide injection, there was a reduction of StAR protein immunoreactivity levels, an accumulation of large lipid droplets in close contact with each other, and an induction of proliferation of the steroidogenic cells. These results indicate a possible role of IGF-II C-peptide in steroidogenic cell function and in induction of steroidogenesis. The detection in this study of IGF-I receptor (IGF-IR) immunoreactivity in frog adrenal glands also indicates that the metabolic and mitogenic effects of IGF-II C-peptide in these glands may occur via the IGF-IR. PMID- 18405945 TI - Photodynamic diagnosis of prostate cancer using 5-aminolevulinic acid--first clinical experiences. AB - OBJECTIVES: To study the feasibility of 5-aminolevulinic-acid (5-ALA)-induced photodynamic diagnosis (PDD) for the evaluation of the surgical margins (SMs) during radical prostatectomy (RP) in patients with prostate cancer (PCa). METHODS: A total of 18 patients with histologically confirmed PCa (Gleason score 4 to 8, prostate-specific antigen 1 to 20 ng/mL) underwent RP. Of the 18 patients, 16 received 5-ALA (20 mg/kg) orally 2 hours before RP, and 2 served as controls without any application of 5-ALA. To study the protoporphyrin IX (PPIX) accumulation after application of 5-ALA, all harvested specimens were investigated by fluorescence microscopy and spectroscopy. In 10 of 16 patients, PDD of the SMs and the prostate was performed during RP using an incoherent light source filtered for efficient fluorescence excitation. RESULTS: In all 16 patients, who had received 5-ALA fluorescence microscopy showed a selective accumulation of PPIX in the PCa cells, and only weak PPIX fluorescence could be detected in benign epithelial cells and none in the adjacent stroma. The 2 patients, who had not received 5-ALA had no PPIX fluorescence in the prostate. Of 10 patients, 8 demonstrated fluorescence-negative and histologically confirmed negative margins during PDD, and 1 each demonstrated a fluorescence-positive SM that was also confirmed by histologic examination and a positive SM that was not confirmed by PPD. CONCLUSIONS: This is the first report of PDD for PCa using 5 ALA. These initial results have demonstrated that PPIX is selectively enhanced in malignant tissue, an essential prerequisite of PDD. Additional studies are warranted to validate these preliminary data and the efficacy of PDD for PCa during RP. PMID- 18405946 TI - Feasibility and economic impact of a clinical pathway for pap test utilization in Gynecologic Oncology practice. AB - OBJECTIVE: To evaluate the feasibility and impact on cost to the U.S. healthcare system of implementing a clinical pathway for Pap test utilization in screening and surveillance of gynecologic cancers in a university-based gynecologic oncology practice. METHODS: Baseline data were collected for Pap test utilization between 1/1/04 and 6/30/05 and prospectively compared to Pap test utilization following the implementation of a clinical pathway (7/1/05 to 5/30/06). The clinical pathway: 1) employed ACOG guidelines for asymptomatic screening of non cancer patients, 2) allowed testing at 4 months intervals for cervical/vaginal cancer surveillance, 3) limited testing for endometrial cancer surveillance to 2 tests/60 months, and 4) eliminated testing as part of ovarian cancer surveillance. Relevant costs were calculated using Medicare charge-to-cost ratios and adjusted to 2006 USD. For statistical analysis, differences in Pap test utilization and cost were evaluated using student's t-test. RESULTS: During the baseline period, 1725 Pap tests were collected from 5605 ambulatory encounters, for a Pap test rate of 30.8% and an annualized cost of $93,759. After implementation of the clinical pathway, 4209 ambulatory encounters yielded an annual Pap test rate of 11% and an annual cost of $35,728 (p<0.0001), a savings of $58,031. In addition, clinical pathway implementation also registered an opportunity cost savings of 180.3 nursing personnel work hours ($4,162). CONCLUSIONS: Reduction in the unnecessary use of Pap testing for asymptomatic screening and surveillance for gynecologic cancers through a straightforward clinical pathway is feasible and offers an opportunity for significant cost savings in gynecologic oncology healthcare expenditure. PMID- 18405947 TI - Ovarian cancer risk is associated with a common variant in the promoter sequence of the mismatch repair gene MLH1. AB - OBJECTIVES: Inherited mutations in the MLH1 gene are associated with a proportion of families with the hereditary non-polyposis colon cancer syndrome (HNPCC). The cardinal features of the syndrome are a predisposition to colon, endometrial and ovarian cancers. Recently, it has been shown that a non-coding polymorphic variant in MLH1 (G>A nt-93) predisposes to colon and endometrial cancer, but with much reduced penetrance. We sought to establish whether or not this polymorphic variant also predisposes to ovarian cancer. METHODS: We genotyped 899 women with invasive ovarian cancer and 931 controls for the G>A nt-93 variant. RESULTS: The presence of the variant was associated with a modest, but highly significant risk of ovarian cancer (OR=1.5; 95% CI 1.3-1.9; p=0.00005). The association was present in cancers of all histologies except clear cell, and in all ethnic groups. CONCLUSIONS: The G>A nt-93 variant of the MLH1 gene is associated with an increased risk of invasive ovarian cancer. PMID- 18405948 TI - Phase II evaluation of CKD-602, a camptothecin analog, administered on a 5-day schedule to patients with platinum-sensitive or -resistant ovarian cancer. AB - BACKGROUND: To evaluate the toxicity and efficacy of a newly developed topoisomerase I inhibitor, CKD-602 in second-line therapy of ovarian cancer. METHODS: We enrolled 24 patients with recurrent ovarian cancer, of median age 54 years (range, 39-64). Eleven patients had measurable lesions on CT scan, and the other 13 had increased serum CA-125 levels. Eighteen patients had platinum sensitive disease (minimum treatment free interval > or =6 months) and 6 had platinum-resistant disease (minimum treatment free interval <6 months). CKD-602 (0.5 mg/m(2)/day) was administered intravenously for 5 days every 3 weeks. The median number of courses per patient was 6 (range, 1 to 12). Response was evaluated by the evaluation of the size of the mass by CT scan and CA-125 response. RESULTS: The overall response rate was 45.0% (9/20), with 4 patients exhibiting partial responses and 5 patients exhibiting 75% CA-125 responses in 20 evaluable patients. Of the 9 responsive patients, 8 were platinum-sensitive (8/15, 53.3%) and 1 was platinum-resistant (1/5, 20.0%). An additional 5 patients showed stable disease, whereas 6 patients exhibited progressive lesions. Of 24 patients, the most common toxicity was hematological, with grades 3 or 4 neutropenia developing in all 24 patients (100%) and in 94 cycles (71.7%). Grade 3 thrombocytopenia developed in 4 patients (16.7%) and 6 cycles (4.6%). None of the patients experienced grades 3 and 4 gastrointestinal toxicities, including nausea, vomiting, and anorexia. CONCLUSIONS: The newly developed topoisomerase I inhibitor, CKD-602, showed activity against both platinum-sensitive and resistant ovarian cancer, with acceptable toxicity. PMID- 18405950 TI - Differential gene expression in the nucleus accumbens with ethanol self administration in inbred alcohol-preferring rats. AB - The current study examined the effects of operant ethanol (EtOH) self administration on gene expression kin the nucleus accumbens (ACB) and amygdala (AMYG) of inbred alcohol-preferring (iP) rats. Rats self-trained on a standard two-lever operant paradigm to administer either water-water, EtOH (15% v/v) water, or saccharin (SAC; 0.0125% g/v)-water. Animals were killed 24 h after the last operant session, and the ACB and AMYG dissected; RNA was extracted and purified for microarray analysis. For the ACB, there were 513 significant differences at the p<0.01 level in named genes: 55 between SAC and water; 215 between EtOH and water, and 243 between EtOH and SAC. In the case of the AMYG (p<0.01), there were 48 between SAC and water, 23 between EtOH and water, and 63 between EtOH and SAC group. Gene Ontology (GO) analysis indicated that differences in the ACB between the EtOH and SAC groups could be grouped into 15 significant (p<0.05) categories, which included major categories such as synaptic transmission, cell and ion homeostasis, and neurogenesis, whereas differences between the EtOH and water groups had only 4 categories, which also included homeostasis and synaptic transmission. Several genes were in common between the EtOH and both the SAC and water groups in the synaptic transmission (e.g., Cav2, Nrxn3, Gabrb2, Gad1, Homer1) and homeostasis (S100b, Prkca, Ftl1) categories. Overall, the results suggest that changes in gene expression in the ACB of iP rats are associated with the reinforcing effects of EtOH. PMID- 18405952 TI - Acute respiratory health effects of air pollution on children with asthma in US inner cities. AB - BACKGROUND: Children with asthma in inner-city communities may be particularly vulnerable to adverse effects of air pollution because of their airways disease and exposure to relatively high levels of motor vehicle emissions. OBJECTIVE: To investigate the association between fluctuations in outdoor air pollution and asthma morbidity among inner-city children with asthma. METHODS: We analyzed data from 861 children with persistent asthma in 7 US urban communities who performed 2-week periods of twice-daily pulmonary function testing every 6 months for 2 years. Asthma symptom data were collected every 2 months. Daily pollution measurements were obtained from the Aerometric Information Retrieval System. The relationship of lung function and symptoms to fluctuations in pollutant concentrations was examined by using mixed models. RESULTS: Almost all pollutant concentrations measured were below the National Ambient Air Quality Standards. In single-pollutant models, higher 5-day average concentrations of NO2, sulfur dioxide, and particles smaller than 2.5 microm were associated with significantly lower pulmonary function. Higher pollutant levels were independently associated with reduced lung function in a 3-pollutant model. Higher concentrations of NO2 and particles smaller than 2.5 microm were associated with asthma-related missed school days, and higher NO2 concentrations were associated with asthma symptoms. CONCLUSION: Among inner-city children with asthma, short-term increases in air pollutant concentrations below the National Ambient Air Quality Standards were associated with adverse respiratory health effects. The associations with NO2 suggest that motor vehicle emissions may be causing excess morbidity in this population. PMID- 18405955 TI - Expression of smooth muscle and extracellular matrix proteins in relation to airway function in asthma. AB - BACKGROUND: Smooth muscle content is increased within the airway wall in patients with asthma and is likely to play a role in airway hyperresponsiveness. However, smooth muscle cells express several contractile and structural proteins, and each of these proteins may influence airway function distinctly. OBJECTIVE: We examined the expression of contractile and structural proteins of smooth muscle cells, as well as extracellular matrix proteins, in bronchial biopsies of patients with asthma, and related these to lung function, airway hyperresponsiveness, and responses to deep inspiration. METHODS: Thirteen patients with asthma (mild persistent, atopic, nonsmoking) participated in this cross-sectional study. FEV(1)% predicted, PC(20) methacholine, and resistance of the respiratory system by the forced oscillation technique during tidal breathing and deep breath were measured. Within 1 week, a bronchoscopy was performed to obtain 6 bronchial biopsies that were immunohistochemically stained for alpha-SM actin, desmin, myosin light chain kinase (MLCK), myosin, calponin, vimentin, elastin, type III collagen, and fibronectin. The level of expression was determined by automated densitometry. RESULTS: PC(20) methacholine was inversely related to the expression of alpha-smooth muscle actin (r = -0.62), desmin (r = 0.56), and elastin (r = -0.78). In addition, FEV(1)% predicted was positively related and deep inspiration-induced bronchodilation inversely related to desmin (r = -0.60), MLCK (r = -0.60), and calponin (r = -0.54) expression. CONCLUSION: Airway hyperresponsiveness, FEV(1)% predicted, and airway responses to deep inspiration are associated with selective expression of airway smooth muscle proteins and components of the extracellular matrix. PMID- 18405957 TI - Airway inflammation: an important marker of control for asthma guidelines? PMID- 18405956 TI - Functional characterization of the atopy-associated gene PHF11. AB - BACKGROUND: Polymorphisms in the plant homeodomain finger protein 11 gene (PHF11) are associated with increased total serum IgE levels, asthma, and severe atopic dermatitis (AD) in children. Although PHF11 includes a plant homeodomain, a motif often found in transcriptional regulators, the function of PHF11 has not been investigated. OBJECTIVE: We sought to test (1) whether PHF11 regulates the transcription of genes involved in allergic disorders and (2) whether polymorphisms in PHF11 predict changes in the expression or function of this gene. METHODS: Microarray analysis was used to examine the expression of PHF11 in different immune cell subsets, and the function of PHF11 was tested by using small interfering RNA-induced knockdown or overexpression of PHF11 in primary CD4+ T cells or Jurkat T cells. Genotype-dependent effects on PHF11 expression were tested by using an allele-specific gene expression, and the transcriptional activity of PHF11 was determined by using luciferase hybrid gene reporter assays and in vitro DNA-binding electromobility shift assays. RESULTS: PHF11 expression was higher in T(H)1 cells relative to that in T(H)2 cells, and knockdown of PHF11 expression reduced expression of the T(H)1-type cytokines IFN-gamma and IL-2. The G-allele of a 3' untranslated region polymorphism associated with AD was correlated with reduced abundance of PHF11 RNA in T(H)1 cells, as well as an increase in a PHF11 isoform lacking exon II. Evidence was also found for a physical and functional interaction between PHF11 and the p65 subunit of nuclear factor kappaB. CONCLUSION: PHF11 is a regulator of T(H)1-type cytokine gene expression. The reduction in PHF11 expression seen with an AD-associated genotype could contribute to the strong T(H)2 responses that characterize many allergic individuals. PMID- 18405958 TI - The extended IL-10 superfamily: IL-10, IL-19, IL-20, IL-22, IL-24, IL-26, IL-28, and IL-29. AB - Cytokines are involved in virtually every aspect of immunity and inflammation. A cascade of responses evolves after cytokine activation, although optimal function might ultimately involve several complementary cytokines. Understanding the function of individual cytokines is complicated because their role can vary depending on the cellular source, target, and phase of the immune response. In fact, numerous cytokines have both proinflammatory and anti-inflammatory potential, with the contrasting outcome observed being determined by the immune cells present and their state of responsiveness to the cytokine. These issues make the study of cytokine biology daunting, particularly so for IL-10 and IL-10 related genes. The IL-10 superfamily is highly pleiotropic. These genes are linked together through genetic similarity and intron-exon gene structure. Significant commonality exists not only through shared receptors but also through conserved signaling cascades. However, its members mediate diverse activities, including immune suppression, enhanced antibacterial and antiviral immunity, antitumor activity, and promotion of self-tolerance in autoimmune diseases. PMID- 18405959 TI - Obesity and asthma: possible mechanisms. AB - Epidemiologic data indicate that obesity increases the prevalence and incidence of asthma and reduces asthma control. Obese mice exhibit innate airway hyperresponsiveness and augmented responses to certain asthma triggers, further supporting a relationship between obesity and asthma. Here I discuss several mechanisms that may explain this relationship. In obesity, lung volume and tidal volume are reduced, events that promote airway narrowing. Obesity also leads to a state of low-grade systemic inflammation that may act on the lung to exacerbate asthma. Obesity-related changes in adipose-derived hormones, including leptin and adiponectin, may participate in these events. Comorbidities of obesity, such as dyslipidemia, gastroesophageal reflux, sleep-disordered breathing, type 2 diabetes, or hypertension may provoke or worsen asthma. Finally, obesity and asthma may share a common etiology, such as common genetics, common in utero conditions, or common predisposing dietary factors. Novel therapeutic strategies for treatment of the obese patient with asthma may result from an increased understanding of the mechanisms underlying this relationship. PMID- 18405960 TI - Effectiveness of a lifestyle intervention and a structured exercise intervention in older adults. AB - OBJECTIVE: To evaluate the effects of a lifestyle intervention and a structured exercise intervention on physical activity in older adults. METHOD: Participants were randomly assigned to a lifestyle intervention (n=60), including an individualized home-based program supported by phone calls, or to a structured intervention (n=60) consisting of three weekly supervised sessions. Results were compared with a control group (n=66). Physical activity was measured with self report questionnaires, pedometers, and accelerometers before the start (pretest), at the end (11 months, posttest), and after 23 months (follow-up). The study took place in Belgium from March 2004 until April 2006. RESULTS: At posttest, both intervention groups had significantly increased their total physical activity compared with the control group. At follow-up, the lifestyle group showed significantly larger increases in active transportation and total steps than the control and structured group respectively. There were no longer significant differences between the structured intervention and the control group. CONCLUSIONS: The structured and lifestyle interventions were equally effective at the end of the intervention. One year after the intervention the lifestyle group maintained a significant increase in physical activity, which highlights the potential of lifestyle programs in the battle against inactivity in older adults. PMID- 18405961 TI - Luminescence of selected dental composites in vitro. AB - OBJECTIVES: The work was motivated by the necessity to evaluate the emission changes of dental composites, as the emission under UV light is one of the important optical parameters of the hard tooth tissue. METHODS: Fulfill extra, Artemis and Durafill VS materials in A3.5 shade were tested. Coffee, tea, red wine and distilled water (blank) were used as staining drinks. The samples were soaked for 7 days. Total luminescence matrices were collected with an interval of 1 nm in emission and 10nm in excitation, in the front-face geometry. RESULTS: The exposure of dental composites to staining drinks caused a reduction in luminescence intensity by up to 40%, the magnitude of the reduction depending on the drink and on the composite material. In the special case of the intrinsically weak fluorescent Durafill VS, the exposure to coffee apparently caused an increase in luminescence intensity due to coffee components adsorbed on the sample surface. SIGNIFICANCE: It is important to improve the emission stability of dental composites. The presently proposed methods are effective in evaluating the emission of dental composites. Emission needs to be tested in addition to color, as the two parameters are uncorrelated. PMID- 18405962 TI - An in vitro biological and anti-bacterial study on a sol-gel derived silver incorporated bioglass system. AB - OBJECTIVES: The purpose of this study was to evaluate the in vitro antibacterial and biological activity of silver-incorporated bioactive glass system SiO2-CaO P2O5-Ag2O (AgBG). The bacteriostatic and bactericidal properties of this new quaternary glass system along with the ternary sol-gel glass system SiO2-CaO-P2O5 (BG) have been studied using Escherichia coli as a test micro-organism. The AGBG system thus appears to be a promising material for dental applications, since similar effects might be produced on a film of bacteria and mucous that grows on the teeth. METHODS: The SiO2-CaO-P2O5-Ag2O and SiO2-CaO-P2O5 glass systems were synthesized by the sol-gel technique and characterized for their physicho chemical properties. The antibacterial activity and biological properties were evaluated by determining the minimum inhibitory concentrations (MICs). Release of Ag+ into the culture medium was measured by inductively coupled plasma (ICP) analysis. RESULTS: The in vitro antibacterial action of the SiO2-CaO-P2O5-Ag2O was compared with that of its ternary counterpart glass system. The concentrations of Ag-bioglass, in the range of 0.02-0.20 mg of Ag-bioglass per millilitre of culture medium, were found to inhibit the growth of these bacteria. The Ag-bioglass not only acts bacteriostatically but it also elicited a rapid bactericidal action. A complete bactericidal effect was elicited in the early stages of the incubation at Ag-bioglass concentration of 20 mg/ml and the ternary glass system had no effect on bacterial growth or viability. The antibacterial action of Ag-bioglass was exclusively attributed to the leaching of Ag+ ions from the glass matrix. SIGNIFICANCE: One of the major advantages of incorporating silver ions into a gel glass system is that the porous glass matrix can allow for controlled sustained delivery of the antibacterial agent to dental material, used even under anaerobic conditions such as deep in the periodontal pocket. This glass system also provides long-term action required for systems which are constantly at risk of microbial contamination. PMID- 18405963 TI - Detoxification mechanisms in shrimp: comparative approach between hydrothermal vent fields and estuarine environments. AB - Hydrothermal vents are extreme deep-sea habitats that, due to their singular features, still intrigue scientific communities. Swift growth rates and profuse biomass of biological communities can be observed, despite of their inherently unstable physical-chemical and toxic conditions, indicating that organisms inhabiting this environment must be well adapted to these inhospitable conditions. The caridean shrimp, Chorocaris chacei, Mirocaris fortunata and Rimicaris exoculata, together with bathymodiolid mussels, dominate the vent fauna along the Mid-Atlantic Ridge (MAR). Crustacean species are widely used as biological indicators of environmental alterations, since they play a key ecological role as planktivorous grazers, epibenthic scavengers or as prey species. The biological consequences of the hydrothermal metal-rich environment in shrimp species are still largely unknown. Therefore, the aim of this study was the determination of the metal levels (Ag, Cd, Cu, Fe, Mn and Zn), metallothioneins (MT) and lipid peroxidation (LPO) in shrimp species collected in Rainbow, Lucky Strike and Menez-Gwen vent sites, in order to evaluate their different adaptation strategies toward metals when compared with two common coastal shrimp species (Palaemon elegans and Palaemonetes varians) from a fairly unpolluted estuarine system in south Portugal (Ria Formosa). Results show significant differences in metal concentrations, MT levels and lipid peroxidation between vent and coastal shrimp and also between shrimp species from the same site. This indicates that biochemical responses in both vent and coastal shrimp are affected not only by the environmental characteristics but also by inter specific differences. Nevertheless, these responses apparently grant a successful adaptation for the survival in a metal-extreme environment. PMID- 18405964 TI - Protective effect of Lycium barbarum polysaccharides on oxidative damage in skeletal muscle of exhaustive exercise rats. AB - The aim of this study was to determine the modulatory effect of Lycium barbarum polysaccharides (LBP) on the oxidative stress induced by an exhaustive exercise. 32 male Wistar rats were taken in the study. The experiment was a 30-day exhaustive exercise program. We determined the lipid peroxidation, glycogen levels, and anti-oxidant enzyme activities in skeletal muscle. The results demonstrated that L. barbarum polysaccharides administration significantly increases glycogen level and anti-oxidant enzyme activities, and decreased malondialdehyde (MDA) level and creatine kinase activities. In conclusion, L. barbarum polysaccharides administration can significantly decrease the oxidative stress induced by the exhaustive exercise. PMID- 18405951 TI - The Inhaled Steroid Treatment As Regular Therapy in Early Asthma (START) study 5 year follow-up: effectiveness of early intervention with budesonide in mild persistent asthma. AB - BACKGROUND: The Inhaled Steroid Treatment as Regular Therapy in Early Asthma (START) study enrolled 7241 patients aged 5 to 66 years with recent-onset, mild persistent asthma to assess early intervention with the inhaled corticosteroid budesonide on long-term asthma control. OBJECTIVE: The open-label phase of the START study was included to determine the effect on lung function and asthma control of adding budesonide to the reference group patients who had not initially received inhaled corticosteroids. METHODS: Patients were randomized to double-blind treatment with budesonide, 200 mug (those aged < 11 years) or 400 mug once daily, or placebo plus the usual asthma therapy for 3 years, after which all patients received 2 years of open-label treatment with budesonide once daily. RESULTS: During the full 5-year study period, postbronchodilator FEV(1) percent predicted decreased, irrespective of randomized treatment during the double-blind phase, by an average of 2.22% (SE, 0.15%). However, patients with inhaled budesonide in the double-blind phase had a significantly lower risk (odds ratio, 0.61; P < .001) of a severe asthma-related event during the full 5-year study period than those in the reference group. Moreover, patients in the reference group used more additional asthma medications during both the open-label and double-blind phases. CONCLUSIONS: In mild persistent asthma early intervention with inhaled budesonide was associated with improved asthma control and less additional asthma medication use. PMID- 18405965 TI - Sustained release of sphingosine 1-phosphate for therapeutic arteriogenesis and bone tissue engineering. AB - Sphingosine 1-phosphate (S1P) is a bioactive phospholipid that impacts migration, proliferation, and survival in diverse cell types, including endothelial cells, smooth muscle cells, and osteoblast-like cells. In this study, we investigated the effects of sustained release of S1P on microvascular remodeling and associated bone defect healing in vivo. The murine dorsal skinfold window chamber model was used to evaluate the structural remodeling response of the microvasculature. Our results demonstrated that 1:400 (w/w) loading and subsequent sustained release of S1P from poly(lactic-co-glycolic acid) (PLAGA) significantly enhanced lumenal diameter expansion of arterioles and venules after 3 and 7 days. Incorporation of 5-bromo-2-deoxyuridine (BrdU) at day 7 revealed significant increases in mural cell proliferation in response to S1P delivery. Additionally, three-dimensional (3D) scaffolds loaded with S1P (1:400) were implanted into critical-size rat calvarial defects, and healing of bony defects was assessed by radiograph X-ray, microcomputed tomography (muCT), and histology. Sustained release of S1P significantly increased the formation of new bone after 2 and 6 weeks of healing and histological results suggest increased numbers of blood vessels in the defect site. Taken together, these experiments support the use of S1P delivery for promoting microvessel diameter expansion and improving the healing outcomes of tissue-engineered therapies. PMID- 18405966 TI - Three-dimensional polymer scaffolds for high throughput cell-based assay systems. AB - Many whole cell-based assays in use today rely on flat, two-dimensional (2D) glass or plastic substrates that may not produce results characteristic of in vivo conditions. In this study, a three-dimensional (3D) cell-based assay platform was established by integrating 3D synthetic polymer scaffolds with standard cell culture dishes and multi-well plates. This technology can be used to feasibly modify any traditional 2D cell-based assay vessels for 3D cell-based assay with currently used high throughput screening (HTS) systems. We examined neural stem (NS) cells' growth profile, morphology, cell-matrix interaction, gene expression and voltage gated calcium channel (VGCC) functionality of this novel 3D assay platform. Our results showed that unlike the NS cells cultured on traditional 2D planar surfaces, cells in 3D scaffolds are more physiologically relevant with respect to in vivo characteristics exhibited by in-vivo surrogates such as neural spheres. This new biomimetic cell-based assay platform may provide a broadly applicable 3D cell-based system for use in drug discovery programs and other research fields. PMID- 18405967 TI - Cucurbitacin B induces differentiation, cell cycle arrest, and actin cytoskeletal alterations in myeloid leukemia cells. AB - Cucurbitacins have long been utilized for their abortifacient and anti inflammatory effects; however, little is known about their mechanism of action. In this study, we have demonstrated robust antiproliferative effects of CuB on various leukemia and lymphoma cell lines, as well as on primary mononuclear bone marrow cells derived from patients with acute myeloid leukemia or myelodysplastic syndrome. Myeloid leukemic cells treated with CuB exhibit significant S-phase cell cycle arrest, enlarged cell size, multinucleation, and enhanced expression of a monocytic- and granulocytic-specific CD11b. Moreover, our data demonstrate that CuB prominently alters the cytoskeletal network of leukemic cells, inducing rapid and improper polymerization of the F-actin network. These encouraging results suggest the appropriateness of clinical trials of cucurbitacins for the treatment of hematopoietic malignancies. PMID- 18405968 TI - Functional analysis of myelodysplastic syndromes-derived mesenchymal stem cells. AB - Two different reports, including one from our own group, have recently demonstrated the presence of severe chromosomal abnormalities in mesenchymal stem cells (MSC) from patients with myelodysplastic syndromes (MDS). In the present study, we have assessed whether such cytogenetic abnormalities result in functional deficiencies in vitro. We found that both normal and MDS MSC showed similar expression patterns of cell adhesion molecules and extracellular matrix proteins. MDS MSC layers showed the capability to differentiate towards adipocytes, chondrocytes and osteoblasts, and supported the growth of early umbilical cord blood progenitors in a co-culture system. Unstimulated MDS MSC secreted more IL-1beta and after treatment with TNFalpha, they secreted more SCF, as compared to their normal counterparts. The present study demonstrates that, in spite of harboring severe chromosomal alterations, most of the functional properties of MDS-derived MSC remain normal, including their ability to support normal hematopoiesis in vitro. PMID- 18405969 TI - Polymorphisms of TNF-alpha and LT-alpha genes in multiple myeloma. AB - Allelic distribution of -308 G>A (TNF 1/2) polymorphism of the TNF-alpha, and the +252 A>G promoter polymorphism of the LT-alpha gene, the 1267 A>G polymorphism of the HSP70-2 gene as well as the -429 T>C promoter polymorphism of the RAGE gene were tested in 94 MM cases and 141 controls. Significantly less MM patients than controls carried the TNF2 allele (p=0.018) and the TNF2-LTA 252G haplotype (p=0.025). The difference was, however, restricted to the females, as well as the relatively young (<69 years) subjects. By contrast, we did not find differences with the other SNPs tested. PMID- 18405970 TI - Lack of antileukemic activity of rapamycin in elderly patients with acute myeloid leukemia evolving from a myelodysplastic syndrome. PMID- 18405971 TI - Iron overload in myelodysplastic syndromes: a Canadian consensus guideline. AB - In December 2005, 11 Canadian hematologists met to develop an evidence-based clinical practice guideline that would address the diagnosis, monitoring, management, and rationale for the treatment of transfusional iron overload in patients with myelodysplastic syndromes (MDS). This Expert Panel consisted of hematologists from across Canada, each with an active practice in a major population centre or a rural area. Based on an extensive literature search and years of clinical experience, their mandate was to address common clinical practice questions, particularly why treat, whom to treat, when to initiate treatment, and how to treat iron overload in patients with MDS. PMID- 18405972 TI - Blood counts at time of complete remission provide additional independent prognostic information in acute myeloid leukemia. AB - Prognostic relevance of blood counts at complete remission (CR) in acute myeloid leukemia (AML) is not clear. To address this issue, we analyzed 891 AML patients in first CR. From the data of randomly selected 446 patients (training set), we first established optimal cutoffs for neutrophil and platelet counts and hemoglobin level at CR in terms of relapse-free survival (RFS). Patients whose counts were higher than each optimal cutoff were shown to have significantly better RFS (p<0.01 for neutrophil and platelets, and p=0.02 for hemoglobin). Then we tested whether these cutoffs were, after accounting for better known prognostic covariates, also predictive of RFS in the remaining 445 patients (validation set). Our data revealed that higher neutrophil count was independently predictive of longer RFS in the validation set (hazard ratio 1.38, p=0.02), as was higher platelet count (hazard ratio 1.35, p=0.04). These findings suggest that blood counts at CR, information readily available, are useful in prognostication in AML. PMID- 18405973 TI - A stereological study of copper toxicity in gills of Oreochromis niloticus. AB - Stereological methods were used to estimate the volumetric density (V(V)) of the filamentar epithelium (FE, 39%), lamellae (L, 28%), central venous sinus (CVS, 14%), central axis (16%), mucous cells (MC, 2%) and chloride cells (CC, 1%) in the gill filament of control Nile Tilapia. The relative volumes of FE and L, and the relative volumes of CVS and central axis, varied inversely under exposure to copper, with high copper toxic levels declanching a chronic defence mechanism that was, nevertheless, overcome, and low copper toxic levels causing adaptation within a moderate acute phase type of response. Copper also induced a decrease of the V(V) (MC, gill filament) due to reduction of surface MC, despite the marked increase of stem MC at chronic exposure to high copper toxic levels. Diminution of the numerical density of filamentar CC was responsible for the decreased V(V) (CC, gill filament), although lamellar CC significantly increased at chronic exposure to low copper toxic levels. The present results demonstrate that cell relative volumes, mean volumes and numerical densities are dependent on the variations of the FE and L, which without a quantitative approach may be misinterpreted, thus stressing the importance of using stereological tools for analyzing histopathological patterns. PMID- 18405974 TI - Sensitivity and specificity of posterior corneal elevation measured by Pentacam in discriminating keratoconus/subclinical keratoconus. AB - PURPOSE: To estimate the sensitivity and specificity of posterior elevation in discriminating keratoconus and subclinical keratoconus from normal corneas. DESIGN: Prospective case-control study. PARTICIPANTS: Seventy-five patients with keratoconus, 25 with subclinical keratoconus, and 64 refractive surgery candidates with normal corneas. METHODS: In one eye of each patient, posterior corneal elevation was measured in the central 5 mm using the Pentacam rotating Scheimpflug camera (Oculus, Wetzlar, Germany). Posterior corneal elevation in keratoconus and subclinical keratoconus were compared with that in normal corneas in separate analyses. Receiver operating characteristic (ROC) curves were used to determine the test's overall predictive accuracy (area under the curve) and to identify optimal posterior corneal elevation cutoff points to maximize sensitivity and specificity in discriminating keratoconus and subclinical keratoconus from normal corneas. Logistic regression was used to support cutoff points identified through ROC curve analysis, and to check for model validity; model goodness-of-fit was estimated using r(2), and its internal validation was by bootstrapping analysis. MAIN OUTCOME MEASURES: Posterior corneal elevation in keratoconus, subclinical keratoconus, and normal corneas. RESULTS: Mean posterior corneal elevation was statistically higher in keratoconus (100.7+/-49.2 microm; P<0.001), and subclinical keratoconus (39.9+/-15.0 microm; P = 0.01) versus normal corneas (19.8+/-6.37 microm). ROC curve analyses showed high overall predictive accuracy of posterior elevation for both keratoconus and subclinical keratoconus (area under the curve 0.99 and 0.93, respectively). Optimal cutoff points were 35 microm for keratoconus and 29 microm for subclinical keratoconus. These values were associated with sensitivity and specificity of 97.3% and 96.9%, respectively, for keratoconus, and 68% and 90.8% for subclinical keratoconus. Similar cutoff points were obtained with logistic regression analysis (38 microm for keratoconus and 32 microm for subclinical keratoconus). The models showed good fit to the data, including after internal validation. CONCLUSIONS: Posterior corneal elevation very effectively discriminates keratoconus from normal corneas. Its efficacy is lower for subclinical keratoconus, and thus data concerning posterior elevation should not be used alone to stratify patients with this condition. PMID- 18405975 TI - Risk factors for relapse of ventilator-associated pneumonia related to nonfermenting Gram negative bacilli: a case-control study. AB - BACKGROUND: The aim of this study was to determine risk factors for relapse of ventilator-associated pneumonia (VAP) related to nonfermenting Gram negative bacilli (NF-GNB). METHODS: This is a retrospective case-control study based on prospectively collected data. Two hundred and seventy six patients with monobacterial VAP related to NF-GNB were eligible. Patients with subsequent superinfection or persistent pulmonary infection were excluded. Patients with relapse of NF-GNB VAP were matched (1:2) with patients without relapse. Matching criteria included the duration of mechanical ventilation before VAP relapse, age+/-5 years, SAPS II at ICU admission+/-5, and the date of admission. Univariate and multivariate analyses were used to determine risk factors for relapse of NF-GNB VAP in cases and controls. RESULTS: Thirty (10%) patients developed a relapse of NF-GNB VAP, 27 (90%) patients were successfully matched with 54 controls. Inappropriate initial antibiotic treatment was the only variable independently associated with relapse of VAP related to NF-GNB (OR [95% CI]=8.1 [2-33], p=0.003). Although ICU-mortality rate was similar in cases and controls (55% vs 72%, p=0.132), the duration of mechanical ventilation and ICU stay were significantly higher in cases than in controls. CONCLUSION: Inappropriate initial antibiotic treatment is independently associated with relapse of VAP related to NF-GNB. PMID- 18405976 TI - Visceral leishmaniasis associated hemophagocytic lymphohistiocytosis--case report and systematic review. AB - BACKGROUND: The clinical features of leishmaniasis overlap with that of hemophagocytic lymphohistiocytosis (HLH) and the diagnosis of visceral leishmaniasis (VL) related HLH can be challenging. OBJECTIVES: To review information available on disease course, treatment, adjunctive therapy used and the outcomes of VL related HLH. METHODS: We describe an illustrative case and review all reported cases of VL associated HLH in the English literature till March 2007. RESULTS: VL associated HLH is rare, with 56 cases reported in the English literature. Clinical features lack discriminating value to recognize VL as the inciting etiology. Bone marrow aspiration (BMA) establishes the diagnosis in 78% of cases but is often negative at onset of the syndrome due to the pauci microbial nature of the disease and patchy involvement. Repeated marrow aspiration, liver biopsy, blood cultures and serology may be required to establish the diagnosis. Liposomal amphotericin is the drug of choice. IVIG may be considered when there is an inadequate response to anti-leishmanial therapy in severe and refractory disease. CONCLUSIONS: VL related HLH is often under recognized because of overlapping clinical features and negative marrow evaluation at onset, leading to high mortality rates. PMID- 18405977 TI - Reliability of mechanomyographic amplitude and mean power frequency during isometric step and ramp muscle actions. AB - This study examined the reliability of mechanomyographic amplitude (MMG RMS) and mean power frequency (MMG MPF) for the vastus lateralis during isometric maximal voluntary contractions (MVCs) and submaximal step and ramp muscle actions of the leg extensors. Nineteen participants (mean+/-S.D. age=24+/-4 yrs) performed two isometric MVCs, 10 randomly ordered isometric step muscle actions at 5, 15, 25, 35, 45, 55, 65, 75, 85, and 95% of MVC, and two 6s isometric ramp muscle actions during 3 trials separated by 3-5 days. The intraclass correlation coefficient (ICC), standard error of the measurement (SEM), and minimal difference (MD) were used for test-retest reliability, while repeated measures ANOVAs were used to quantify systematic variability. Systematic variability was only present across the 3 trials at 5% MVC during the step muscle action for MMG RMS and at 35% of MVC during the ramp muscle action for MMG MPF. The ICCs ranged from 0.39 to 0.89 and 0.36 to 0.80, while the SEM (% of mean) ranged from 14.9 to 33.9% and 6.9 to18.9% for MMG RMS and MMG MPF, respectively. Thus, MMG RMS and MMG MPF demonstrated reliability that was comparable to that of the surface electromyographic signal. PMID- 18405978 TI - TrackFly: virtual reality for a behavioral system analysis in free-flying fruit flies. AB - Modern neuroscience and the interest in biomimetic control design demand increasingly sophisticated experimental techniques that can be applied in freely moving animals under realistic behavioral conditions. To explore sensorimotor flight control mechanisms in free-flying fruit flies (Drosophila melanogaster), we equipped a wind tunnel with a Virtual Reality (VR) display system based on standard digital hardware and a 3D path tracking system. We demonstrate the experimental power of this approach by example of a 'one-parameter open loop' testing paradigm. It provided (1) a straightforward measure of transient responses in presence of open loop visual stimulation; (2) high data throughput and standardized measurement conditions from process automation; and (3) simplified data analysis due to well-defined testing conditions. Being based on standard hardware and software techniques, our methods provide an affordable, easy to replicate and general solution for a broad range of behavioral applications in freely moving animals. Particular relevance for advanced behavioral research tools originates from the need to perform detailed behavioral analyses in genetically modified organisms and animal models for disease research. PMID- 18405979 TI - Are hypomanics the happier normals? AB - BACKGROUND: Not much is known about hypomanic states in subjects free of major and minor depressive mood disorders. Our aim was to identify and characterise a group of such "pure" hypomanics in relation to a normal control group. METHODS: Data were obtained from the Swiss Zurich study, a stratified epidemiological sample of young adults from the general population, followed from age 20 to 40. "Pure" hypomania was defined as a period of increased activity and decreased need for sleep with consequences (e.g. legal trouble or reactions by others). Minor and major mood disorders were excluded. RESULTS: Twenty-three subjects were identified as pure hypomanics. They overlapped minimally with and were clearly different from subjects with DSM-IV defined hypomanic episodes, most of whom had a bipolar disorder. Pure hypomanics were characterised by physical and social overactivity, elevated and irritable mood, as well as increases in extraversion, sexual interest, and risk-taking behaviors. They had higher monthly incomes and were more often married than controls. Subjective distress due to hypomanic symptoms was virtually absent. Quality of life and treatment rates for mood and anxiety were not different from controls, although sleep disturbances, substance abuse and binge eating were more frequent. LIMITATIONS: The subsample identified was small. Due to the focus of the study on pathology, some positive aspects of hypomania may have been missed. CONCLUSIONS: The existence of a group of pure hypomanics presenting a mixed picture of clinically relevant and irrelevant characteristics supports the concept of a continuum from normal to pathological mood states. PMID- 18405980 TI - Neuron-specific enolase antibodies in patients with sudden acquired retinal degeneration syndrome. AB - Sudden acquired retinal degeneration syndrome (SARDS) is a disease characterised by sudden and bilateral vision loss of dogs. Previous studies failed to identify the underlying cause [Mattson, A., Roberts, S.M., Isherwood, J.M.E., 1992. Clinical features suggesting hyperadrenocorticism associated with sudden acquired retinal degeneration syndrome in a dog. J. Am. Anim. Hosp. Assoc. 28, 199-202; Van der Woerdt, A., Nasisse, M.P., Davidson, M.G., 1991. Sudden acquired retinal degeneration in the dog: clinical and laboratory findings in 36 cases. Prog. Vet. Comp. Ophthamol. 1, 11-18] and earlier investigations about the occurrence of anti-retinal antibodies in SARDS patients showed inconsistent results. To provide a novel approach to those findings we designed a more detailed study. Autoantibodies of SARDS patients and normal controls were tested against the purified autoantigens S-antigen and cellular retinaldehyde binding protein (CRALBP) that play a role in human autoimmune uveitis. Next we tested the autoantibody binding pattern to whole retinal lysate. No difference in the incidence of autoantibodies could be found between SARDS patients and healthy controls while testing the well-known autoantigens S-antigen and CRALBP. Potential novel, yet unknown autoantigens were identified by a screening test using the retinal proteome as an autoantigenic source. In SARDS patients and normal controls, several retinal proteins were bound by IgG antibodies, but one band was strongly marked by SARDS patients. That band was excised, subjected to mass spectrometry (matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF/TOF)) and identified as neuron-specific enolase. Binding of the IgG autoantibodies of SARDS-affected dogs to this protein was verified using purified NSE, revealing 25% of NSE autoantibody-positive SARDS patients and 0% of negative controls. Our findings indicate that at least some dogs with SARDS have autoantibodies against NSE, although it is unclear whether these play a causative role in SARDS or whether they are the result of retinal destruction by another mechanism. PMID- 18405981 TI - Effects of physiologic agonists on canine whole blood flow cytometry assays of leukocyte-platelet aggregation and platelet activation. AB - Platelets play a role in both the innate and adaptive immune systems. Methods for detecting activated platelets and leukocyte-platelet aggregates (LPAs) are useful for basic and applied research concerning the role of platelets in inflammation and immune disorders. The aim of the study was to develop flow cytometric assays for detection of platelets binding to monocytes and neutrophils and for activated platelets in canine whole blood and to investigate the effect of physiologic agonists. Citrate anticoagulated whole blood was incubated with monoclonal antibodies against CD14 and CD61 for detection of LPAs, and the effect of various agonists was investigated. For detection of activated platelets, whole blood was incubated with monoclonal antibodies against CD62P and against a receptor-induced binding site on fibrinogen (CAP1) with CD61 as a platelet identifier. Isotype controls were prepared in parallel. The individual physiologic agonists ADP, collagen and epinephrine increased LPAs, CD62P and CAP1 binding only modestly. However, combinations of agonists gave more substantial increases. A dose response relationship was seen using alpha- and gamma-thrombin, and ADP as agonists. In conclusion, we have developed flow cytometry assays to measure LPAs and platelet activation in canine whole blood, and have explored the effect of various physiologic agonists at different concentrations. PMID- 18405982 TI - Chronically stimulated mouse invariant NKT cell lines have a preserved capacity to enhance protection against experimental tumor metastases. AB - In pre-clinical models, CD1d restricted invariant Natural Killer T (iNKT) cells play a pivotal role in natural anti-tumor immune responses, mainly by trans activating cells of both the innate and adaptive arms via swift and potent cytokine secretion. We have previously reported that patients with a severely reduced circulating iNKT cell pool have a poor clinical response to radio therapy of head and neck squamous cell carcinoma. Therefore, these patients might benefit from an immunotherapeutic approach aimed at the increase of circulating levels of iNKT cells. Furthermore, we have generated both human and mouse iNKT cell lines, and demonstrated that they had retained the capacity to release both Th1 and Th2 type cytokines even after long-term in vitro expansion using alpha galactosylceramide (alphaGalCer) pulsed dendritic cells (DC). Here, we establish, in a pre-clinical tumor model that the large scale long lived polyclonal iNKT cell lines we generated have a preserved capacity to evoke an in vivo cytokine storm upon adoptive transfer, independently of supplemental alphaGalCer administration. This results in an augmented NK cell mediated protection against B16.F10 experimental lung metastases in vivo. These findings underscore the potential of autologous adoptive transfer of ex vivo expanded iNKT cells as a strategy to enhance immunotherapeutic modalities for the treatment of cancer patients. PMID- 18405983 TI - Toxic effects of an oil spill on fish early life stages may not be exclusively associated to PAHs: studies with Prestige oil and medaka (Oryzias latipes). AB - Polycyclic aromatic hydrocarbons (PAHs) are assumed to be the primary determinant of oil petroleum toxicity. Since the PAH content in Prestige oil was relatively high, we investigated the effects of different oil fractions (crude or weathered oil -0.05 to 50 g/L, and shaken or sonicated water accommodated fractions, WAFs, 25-100%, v/v) on the embryo-larval development of medaka (Oryzias latipes). Concentrations of summation operator16PAHs analyzed in the incubation medium were highest in the shaken WAF followed by the crude oil, the sonicated WAF and the weathered oil. Both oils (> or =0.25 g/L) induced developmental abnormalities whereas no significant effects were seen in the WAF exposures. In vivo morphometric analysis of the surface of the gallbladder during advanced embryo organogenesis (192 h post-fertilization, hpf) revealed significant dilation in both WAF exposures (>3 x 10(4) microm(2) at > or =25%, v/v, compared to <1.7 x 10(4) microm(2) at 0%, v/v) followed by the crude oil (>2.2 x 10(4) microm(2) at > or =0.05 g/L). Fluorescent aromatic compounds were observed in the gallbladder and the yolk sac of 168-hpf embryos exposed to all oil fractions. Results suggest the presence of components in both oils capable of penetrating the chorion and inducing a toxicity not observed in the WAFs. Hence, the hazard and risk assessment of Prestige oil should not be based solely on the presence of PAHs since proximity or direct contact may induce toxicity not associated exclusively to these compounds. This research offers a new hypothesis for explaining the reported biological observations, which could be correlated to direct oil exposure rather than the traditional mechanism of waterborne PAH exposure. Further research is needed to identify those oil components responsible for toxicity. PMID- 18405984 TI - Immuno stimulating glycophosphosphingolipid antigen from Leishmania donovani is recognized by visceral leishmaniasis patient sera. AB - Surface antigens on Leishmania promastigotes and infected macrophages are obvious targets in immunoprophylaxis for leishmanial infection. The glycophosphosphingolipid (GSPL) antigen isolated from Leishmania donovani surface membrane was recognized by sera from patients with visceral leishmaniasis. GSPL was also expressed on the membrane of parasite-infected macrophages. The effect of GSPL on the production of reactive oxygen species (ROS) and reactive nitrogen intermediates (RNI) was studied using the macrophage cell line J774.1. In addition, induction of IFNgamma, IL4, IL10, IL12 secretion in presence of GSPL was investigated in PBMC from normal individuals. ROS and RNI in addition to IFNgamma and IL12 were induced by GSPL. Though there was a moderate induction of IL10, there was very little induction of the Th2 cytokine IL4. GSPL also induced blood cells to proliferate. The data suggests that this functionally important antigen of L. donovani may be used as a candidate vaccine. PMID- 18405985 TI - Correlative light/electron microscopy for the investigation of microbial mats from Black Sea Cold Seeps. AB - In several fields of cell biology, correlative microscopy is applied to compare the structure of objects at high resolution under the electron microscope with low resolution light microscopy images of the same sample. It is, however, difficult to prepare samples and marker systems that are applicable for both microscopic techniques for the same specimen at the same time. In our studies, we used microbial mats from Cold Seep communities for a simple and rapid correlative microscopy method. The mats consist of bacterial and archaeal microorganisms, coupling reverse methanogenesis to the reduction of sulfate. The reverse methanogenic pathway also generates carbonates that precipitate inside the mat and may be the main reason for the formation of a microbial reef. The mat shows highly differentiated aggregates of various organisms, tightly interconnected by extracellular polysaccharides. In order to investigate the role of EPS as adhesive mucilage for the biofilm and as a precipitation matrix for carbonate minerals, samples were embedded in a hydrophilic resin (Lowicryl K4 M). Sections were suitable for light as well as electron microscopy in combination with lectins, either labeled with a fluorescent marker or with colloidal gold. This allows lectin mapping at low resolution for light microscopy in direct comparison with a highly resolved electron microscopic image. PMID- 18405986 TI - Can protons improve SBRT for lung lesions? Dosimetric considerations. AB - BACKGROUND AND PURPOSE: The aim of the present study was to investigate potential dosimetric benefits of proton therapy for hypofractionated stereotactic body radiotherapy (SBRT). MATERIALS AND METHOD: Twelve patients undergoing hypofractionated SBRT at the Medical University Vienna were selected. Passively scattered protons (PT) and intensity modulated proton therapy (IMPT) were evaluated against a conformal photon technique (3D-CRT), assuming a fractionation of 3x15Gy, prescribed to the 65% isodose. For all treatment techniques shallow breathing with abdominal compression (SB+AC) was compared with a deep inspiration breath hold technique (DIBH). Treatment planning was done with XiO (CMS, USA). Target conformity, dose-volume histograms (DVH) and various associated dosimetric parameters were considered for the planning target volume (PTV), lung, heart and esophagus. RESULTS: For both breathing conditions conformity indices were very similar. They were between 0.75 and 0.78 for IMPT and 3D-CRT and around 0.55 for PT using 2-3 beams. Irrespective of treatment modality, DVHs for the ipsilateral lung were improved with the DIBH technique. For the PT technique, the 2Gy isodose (V2Gy) covered on average 7-9% less lung volume compared to 3D-CRT, for IMPT this reduction was more than 10%. Volumes covered the 4 and 6Gy isodoses were 2-4% smaller for IMPT, but very similar for PT and 3D-CRT. Both proton techniques achieved full sparing of the contralateral lung and superior sparing of the heart. Maximum doses to the heart and esophagus were on average around 3Gy for 3D CRT and almost 0Gy for both proton techniques. For 3D-CRT average V2Gy values for the heart could be reduced from 64% in shallow breathing to 34% in DIBH. V2Gy for protons was negligible. CONCLUSIONS: Only small dosimetric differences were found between photons and protons for SBRT of lung lesions. Whether these small dosimetric benefits translate in reduced side effects or have the potential to improve local control rates remains to be demonstrated in clinical studies. PMID- 18405987 TI - A phase I trial of definitive chemoradiotherapy with docetaxel, cisplatin, and 5 fluorouracil (DCF-R) for advanced esophageal carcinoma: Kitasato digestive disease & oncology group trial (KDOG 0501). AB - BACKGROUND AND PURPOSE: A dose-escalation study of docetaxel combined with cisplatin, 5-fluorouracil, and concurrent radiotherapy (DCF-R) was performed to determine the optimal dose in patients with advanced esophageal carcinoma. PATIENTS AND METHODS: A total of 19 patients who had previously untreated thoracic esophageal carcinoma with T4 tumors and/or M1 lymph-node metastasis were studied. The Patients received an infusion of docetaxel (levels 1, 2, 3, 2.5: 20, 30, 40, 35 mg/m(2)) and an infusion of cisplatin (40 mg/m(2)) on days 1, 15, 29, and 43 plus a continuous infusion of 5-fluorouracil (400mg/m(2)/day) on days 1-5, 15-19, 29-33, and 43-47. And patients received 61.2 Gy/34 fractions/7 weeks of concurrent radiotherapy. RESULTS: Dose-limiting toxicities (DLTs) were febrile neutropenia and grade 4 leukopenia lasting 3 days. DLT occurred in 2 of 6 patients at level 2, 3 of 4 patients at level 3, and 2 of 6 patients at level 2.5. The main toxicities were myelotoxicity and esophagitis. The overall response rate was 89.5%, including a complete response rate of 42.1%. CONCLUSIONS: The maximum-tolerated dose was level 3, because 50% or more of the patients had DLTs. Therefore, level 2.5 was recommended for phase II studies. This regimen was tolerable and highly active. PMID- 18405988 TI - Changes in muscle and joint coordination in learning to direct forces. AB - While it has been suggested that bi-articular muscles have a specialized role in directing external reaction forces, it is unclear how humans learn to coordinate mono- and bi-articular muscles to perform force-directing tasks. Participants were asked to direct pedal forces in a specified target direction during one legged cycling. We expected that with practice, performance improvement would be associated with specific changes in joint torque patterns and mono- and bi articular muscular coordination. Nine male participants practiced pedaling an ergometer with only their left leg, and were instructed to always direct their applied pedal force perpendicular to the crank arm (target direction) and to maintain a constant pedaling speed. After a single practice session, the mean error between the applied and target pedal force directions decreased significantly. This improved performance was accompanied by a significant decrease in the amount of ankle angular motion and a smaller increase in knee and hip angular motion. This coincided with a re-organization of lower extremity joint torques, with a decrease in ankle plantarflexor torque and an increase in knee and hip flexor torques. Changes were seen in both mono- and bi-articular muscle activity patterns. The mono-articular muscles exhibited greater alterations, and appeared to contribute to both mechanical work and force directing. With practice, a loosening of the coupling between bi-articular thigh muscle activation and joint torque co-regulation was observed. The results demonstrated that participants were able to learn a complex and dynamic force directing task by changing the direction of their applied pedal forces through re organization of joint torque patterns and mono- and bi-articular muscle coordination. PMID- 18405989 TI - Sensorimotor synchronization with adaptively timed sequences. AB - Most studies of human sensorimotor synchronization require participants to coordinate actions with computer-controlled event sequences that are unresponsive to their behavior. In the present research, the computer was programmed to carry out phase and/or period correction in response to asynchronies between taps and tones, and thereby to modulate adaptively the timing of the auditory sequence that human participants were synchronizing with, as a human partner might do. In five experiments the computer's error correction parameters were varied over a wide range, including "uncooperative" settings that a human synchronization partner could not (or would not normally) adopt. Musically trained participants were able to maintain synchrony in all these situations, but their behavior varied systematically as a function of the computer's parameter settings. Computer simulations were conducted to infer the human participants' error correction parameters from statistical properties of their behavior (means, standard deviations, auto- and cross-correlations). The results suggest that participants maintained a fixed gain of phase correction as long as the computer was cooperative, but changed their error correction strategies adaptively when faced with an uncooperative computer. PMID- 18405990 TI - Kinetics of hula hooping: an inverse dynamics analysis. AB - This paper involved a biomechanical analysis of lower limb joint coordination during hula hooping. A lower extremity inverse dynamics model that incorporated kinematic input and force platform data was developed to compute the angular velocities, moments about and powers produced at the lower extremity joints. The abductor moments and powers were discovered to be paramount in maintaining hoop oscillations, as demonstrated consistently in the three study participants. However, hula hooping was demonstrated to be variable in terms of the involvement of flexor and extensor moments and powers of the ankle, knee and hip joints, resulting in the adoption of varying strategies by each of the three participants. PMID- 18405991 TI - Thermal resistance of Salmonella serovars isolated from raw, frozen chicken nuggets/strips, nugget meat and pelleted broiler feed. AB - Raw, frozen chicken nuggets/strips available at retail and prepared at home before consumption have been identified as a significant risk factor in contracting food-borne salmonellosis. Cases of salmonellosis from consumption of these products may be due, in part, to Salmonella strains originating in broiler feed. In this study the thermal resistances of Salmonella strains isolated from chicken nuggets and strips, chicken nugget/strip meat and broiler feed were determined to assess whether they exhibited enhanced thermal resistance. Thermal resistances (D- and z- values) of 7 cocktails (25 isolates, 4 serovars) were determined in commercially prepared irradiation-treated chicken nugget/strip meat blend, and heated in a constant temperature waterbath. The thermal resistances found were lower than those reported for similar strains in the literature. D values ranged from 6.93 to 0.12 min at 55 and 62 degrees C respectively, with z values from 4.10 to 5.17 degrees C. Two strains of S. Enteritidis separately isolated from pelleted feed and chicken nugget meat blend, with indistinguishable geno- and phenotypes, had lower (and probably identical) thermal resistances than the other isolates. Results indicated that the strains of Salmonella isolated from raw, frozen chicken nuggets/strips and pelleted broiler feed did not exhibit unusually high thermal resistance, and that normal heating (71 degrees C) prior to consumption should eliminate these organisms from chicken nuggets/strips. PMID- 18405992 TI - Quantitative modeling for risk assessment of Vibrio parahaemolyticus in bloody clams in southern Thailand. AB - A risk assessment of Vibrio parahaemolyticus in bloody clams (Anadara granosa) consumed in southern Thailand was conducted. This study estimated the prevalence and concentration of pathogenic V. parahaemolyticus in bloody clams at harvest and retail stages; and during this process, methods to detect the total and pathogenic V. parahaemolyticus were investigated. Consumption of bloody clams and cooking efficiency were studied using interviews and on-site observation of consumers. A beta-Poisson dose-response model was used to estimate probability of illness applying estimation methods for the most likely parameter values presented by USFDA. Microbial and behavioral data were analyzed by developing a stochastic model and the simulation gave a mean number of times a person would get ill with V. parahaemolyticus by consuming bloody clams at 5.6 x 10( 4)/person/year. Sensitivity analysis demonstrated the fraction of people who did not boil the clams properly was the primary factor in increasing risk. This study serves as an example of how a microbiological risk assessment with limited data collection and international cooperation leads to valuable local insight. PMID- 18405993 TI - Monitoring of transcriptome and proteome profiles to investigate the cellular response of E. coli towards recombinant protein expression under defined chemostat conditions. AB - The use of strong promoter systems for recombinant protein production generates high product yields, but also overburdens the host cell metabolism and compromises production. Escherichia coli has highly developed regulatory pathways that are immediately responsive to adverse conditions. To gain insight into stress response mechanisms and to detect marker genes and proteins for stress specific monitoring time course analysis of controlled chemostat cultivations was performed using E. coli total microarray and difference gel electrophoresis (Ettan DIGE). In order to detect differences and consistencies of stress response as well as the impact of the inducer isopropyl-beta-d-thiogalactopyranosid on cells, expression of two recombinant proteins (hSOD and GFPmut3.1) was investigated. Genes involved in aerobic metabolism under control of the ArcB/ArcA two component system were found to be down-regulated, and the interplay of the psp operon, ArcA system and guanosine tetraphosphate is suggested to be involved in stress regulatory mechanisms. A distinct impact of the two recombinant proteins was observed, particularly on levels of known stress regulatory genes and proteins, as well as on the response associated with ArcA and psp. Altogether, 62 genes as well as seven proteins showed consistent expression levels due to recombinant gene expression, and are therefore suggested to be appropriate monitoring targets. PMID- 18405994 TI - Enzymatic hydrolysis of PTT polymers and oligomers. AB - Oligomers and polymers (film, fabrics) of the linear aromatic polyester poly(trimethylene terephthalate) (PTT) were treated with polyesterases from Thermomyces lanuginosus, Penicillium citrinum, Thermobifida fusca and Fusarium solani pisi. The cutinase from T. fusca was found to release the highest amounts of hydrolysis products from PTT materials and was able to open and hydrolyse a cyclic PTT dimer according to RP-HPLC-UV detection. In contrast, the lipase from T. lanuginosus also showed activity on the PTT fibres and on bis(3-hydroxypropyl) terephthalate (BHPT) but was not able to hydrolyse the polymer film, mono(3 hydroxypropyl) terephthalate (MHPT) nor the cyclic dimer of PTT. As control enzymes inhibited with mercury chloride were used. Surface hydrophilicity changes were investigated with contact angle measurements and the degree of crystallinity changes were determined with DSC. PMID- 18405995 TI - Role of a geminivirus AV2 protein putative protein kinase C motif on subcellular localization and pathogenicity. AB - Virus-derived genes or genome fragments are increasingly being used to generate transgenic plants with resistance to plant viruses. There is need to rapidly investigate these genes in plants using transient expression prior to using them as transgenes since they may be pathogenic to plants. In this study, we investigated the AV2 protein encoded by East African cassava mosaic Cameroon virus, a virus associated with a cassava disease epidemic in western Africa. For subcellular localization, AV2 was fused to the yellow fluorescent protein (YFP) and expressed in Nicotiana benthamiana. Confocal analyses showed that AV2-YFP localizes mainly in the cytoplasm. Because it overlaps with the coat protein gene and therefore could be used to generate transgenic plants for resistance to geminiviruses, we investigated its pathogenesis in N. benthamiana by using the Potato virus X (PVX) vector. The chimeric virus PVX-AV2 induced a mild mottling in infected plants and was shown to suppress virus-induced gene silencing (VIGS). Using point mutations, we show here that AV2 pathogenicity is dependent on a conserved putative protein kinase C (PKC) phosphorylation motif. Because of its pathogenicity and ability to suppress RNA silencing, AV2 transgenic plants will less likely provide a control to geminiviruses, indeed it may weaken the resistance of the plant. We therefore suggest the use of the AV2 putative PKC mutants to generate transgenic plants. PMID- 18405996 TI - DC-SIGN enhances infection of cells with glycosylated West Nile virus in vitro and virus replication in human dendritic cells induces production of IFN-alpha and TNF-alpha. AB - The recent introduction of West Nile virus (WNV) into the Western hemisphere resulted in significant human outbreaks causing disease of variable severity. Previous studies classified WNV into two major lineages (L1 and L2) that differ in their virulence. Since most L1 strains are glycosylated, we investigated the role of dendritic cell-specific ICAM-3-grabbing non-integrin (DC-SIGN) in infection efficiency of glycosylated WNV strains. We showed that glycosylated strains, in contrast to non-glycosylated strains, infected DC-SIGN expressing cells more efficiently than DC-SIGN negative cells. Furthermore, WNV can productively infect cultured human dendritic cells (DCs) and infection of dendritic cells with the glycosylated WNV-NY99 L1 strain induced production of significantly more TNF-alpha and IFN-alpha in cultured DC, than infection with the non-glycosylated B956 L2 strain. Together, these results indicate that DC SIGN enhances infection of cells by WNV glycosylated strains, which may at least in part explain the higher pathogenicity of glycosylated L1 strains versus most non-glycosylated L2 strains. PMID- 18405997 TI - Sequence analysis of a reovirus isolated from the winter moth Operophtera brumata (Lepidoptera: Geometridae) and its parasitoid wasp Phobocampe tempestiva (Hymenoptera: Ichneumonidae). AB - A reovirus was isolated from Operophtera brumata (ObRV) and its parasitoid wasp Phobocampe tempestiva. Each of the 10 dsRNA genome segments of ObRV was sequenced and shown to contain a single open reading frame (ORF). Conserved motifs ([+ve] 5'-AAATAAA ...(G)/(T)AGGTT-3') were found at the termini of each segment, with the exception of Seg-6 and Seg-8, where the 5' termini were 5'-AACAAA...-3'. The putative proteins encoded by each segment were compared with those of other members of the family Reoviridae. Phylogenetic comparisons to published sequences for the RNA-dependent RNA polymerase genes from other reoviruses indicated that ObRV is most closely related to members of the genus Cypovirus. However, unlike the cypoviruses, ObRV has a double-layered capsid structure. When the protein encoded by ObRV Seg-10 was expressed (by inserting the open reading frame into a baculovirus expression vector) no 'occlusion bodies' were observed in the recombinant baculovirus infected insect cell cultures. This suggests that unlike the cypoviruses, Seg-10 of ObRV does not contain a polyhedrin gene. Further phylogenetic comparisons also identified relationships between Seg-2 and Seg-10 of ObRV, and genes of Diadromus pulchellus Idnoreovirus 1 (DpIRV1), suggesting that ObRV represents a new species from the genus Idnoreovirus. PMID- 18405998 TI - Lytic infection with vaccinia virus activates caspases in a Bcl-2-inhibitable manner. AB - Vaccinia virus (VV) is considered to cause lytic infection of most cells, with lysis being regarded equivalent to necrosis. Activation of caspases has not been associated with necrosis. However, we observed the activation and activity of caspases in epithelial cells HeLa G and BSC-40 lytically infected with VV. Using three different flow-cytometric approaches, we characterized the distinct stages of caspase cascade in VV-infected cells: a cleaved, activated form of caspases detected using a fluorescent pan-caspase inhibitor; caspase activity assayed by cleavage of a non-fluorescent substrate into a fluorescent product; caspase specific cleavage of death substrates characterized by a fluorescent antibody detecting a neo-epitope in cytokeratin-18. All of these approaches yielded an increased fluorescent signal in VV-infected cells compared to mock-infected controls. Additionally, the signal was decreased by the expression of Bcl-2. The cleavage of cytokeratin-18 was confirmed by western blotting, but another key protein involved in apoptosis, PARP, was not cleaved in VV-infected lytic cells. The necrotic phenotype of the cells was confirmed by increased cell membrane permeability and/or decreased mitochondrial membrane potential. In conclusion, our data suggest that VV infection of the epithelial cells HeLa G and BSC-40 initiates the apoptotic program, however, apoptosis is not completed and switches into necrosis. PMID- 18405999 TI - In vivo and in vitro studies of GAD-antibody positive subjects with Type 2 diabetes: A distinct sub-phenotype. AB - The purpose of this study was to determine if immune mechanisms in GAD positive patients' contribute to the pathogenesis of a specific sub-type of Type 2 diabetes. GAD positive (n=8) and GAD negative (n=8) subjects diagnosed with Type 2 diabetes were matched for age, gender, body mass index, duration of diabetes and glycaemic control. All subjects underwent an insulin-modified frequently sampled intravenous glucose tolerance test to measure insulin sensitivity and insulin secretory function with minimal model analysis. In addition, BRIN-BD11 clonal beta-cells were supplemented with patients' sera to determine basal and alanine-stimulated insulin secretion and terminal complement complex (TCC) formation. Both groups were severely insulin resistant (0.56+/-0.17 vs. 0.99+/ 0.3310(-4)min(-1)/(microUml(-1)) for GADneg and GADpos, respectively) but the GAD negative subjects had a higher basal (87+/-11 vs. 58+/-14pmoll(-1), p<0.05) and glucose-stimulated insulin secretion (DeltaAUCins 0.96+/-0.12 vs. 0.60+/ 0.12pmol/(l(-1)min), p<0.05). In vivo measures of insulin secretion were negatively correlated with TCC formation, independent of antibody status. In conclusion, GAD positive subjects initially diagnosed with Type 2 diabetes are unable to compensate for insulin resistance due to more pronounced beta-cell impairment. TCC formation may be partly responsible for the insulin secretory dysfunction associated with this specific sub-type of Type 2 diabetes. PMID- 18406000 TI - Effect of rosuvastatin on capillary filtration of albumin and blood pressure in rats with streptozotocin-induced diabetes. AB - An increase in capillary filtration of albumin (CFA) is well demonstrated in diabetes. Statins may exert a protective effect against endothelial dysfunction. The aim of this study was to test whether rosuvastatin may prevent the increase in peripheral CFA in diabetic rats and the role of blood pressure lowering. Rats with streptozotocin-induced diabetes were randomized to receive either rosuvastatin 20mg/kg/d (group R) or both rosuvastatin 20mg/kg/d and mevalonate 20mg/kg/d (group RM) or no treatment (group U). CFA index was measured on a limb by a non-invasive isotopic test using technetium-labelled albumin, at three time points: at mean age of 3 months, before treatment; at 5 and 8 months, i.e. after 2 and 5 months of treatment. At 3 months, interstitial albumin retention (AR) was markedly increased in the 3 groups. From 3 to 5 months, AR increased significantly in group U, decreased in group R and in group RM. At 5 and 8 months, AR was significantly lower in groups R and RM than in group U. Systolic blood pressure (SBP) was measured at 8 months and was significantly lower in group R than in group U and RM. At 8 months, serum cholesterol levels were not different between the three groups whereas triglycerides were significantly lower in groups R and RM than in group U. In conclusion, in diabetic rats rosuvastatin prevents the increase in peripheral CFA and induces a decrease in blood pressure. The beneficial effect of rosuvastatin on endothelial function does not seem to result from blood pressure reduction nor lipid lowering effects. PMID- 18406001 TI - Gestational diabetes has no additional effect on plasma thrombin-activatable fibrinolysis inhibitor antigen levels beyond pregnancy. AB - Pregnancy is a prothrombotic condition with increased levels of several circulating coagulation factors. Decreased fibrinolytic activity has been shown in gestational diabetes. Gestational diabetes has been found to be associated with higher plasma plasminogen activator inhibitor-1 (PAI-1) antigen levels than normal pregnancy. The aim of the present study is to investigate the effect of gestational diabetes on plasma thrombin-activatable fibrinolysis inhibitor (TAFI) antigen levels. Plasma TAFI and PAI-1 antigen levels were measured in 26 pregnant women with gestational diabetes, 25 pregnant women with normal glucose tolerance, and age-matched 24 non-pregnant women with no history of gestational diabetes. Increased plasma TAFI antigen levels were found in pregnant women compared to non pregnant controls. However, no statistically significant difference in TAFI antigen levels was observed between women with gestational diabetes and pregnant controls. Plasma PAI-1 antigen levels were higher in gestational diabetes than pregnant and non-pregnant controls. Our study revealed that pregnancy was associated with elevated plasma TAFI antigen levels. However, no additional effect of gestational diabetes was found on plasma TAFI antigen levels beyond pregnancy. We suggest that pregnancy is associated with enhanced coagulation and impaired fibrinolysis. Despite increased PAI-1 antigen levels associated with gestational diabetes, the effect of gestational diabetes on TAFI antigen levels is lacking. PMID- 18406002 TI - Methods and implementation of a central biosample and data management in a three centre clinical study. AB - In our report we describe concept, strategies and implementation of a central biosample and data management (CSDM) system in the three-centre clinical study of the Transregional Collaborative Research Centre "Inflammatory Cardiomyopathy - Molecular Pathogenesis and Therapy" SFB/TR 19, Germany. Following the requirements of high system resource availability, data security, privacy protection and quality assurance, a web-based CSDM was developed based on Java 2 Enterprise Edition using an Oracle database. An efficient and reliable sample documentation system using bar code labelling, a partitioning storage algorithm and an online documentation software was implemented. An online electronic case report form is used to acquire patient-related data. Strict rules for access to the online applications and secure connections are used to account for privacy protection and data security. Challenges for the implementation of the CSDM resided at project, technical and organisational level as well as at staff level. PMID- 18406003 TI - Recent developments in enzyme-catalyzed ring-opening polymerization. AB - An exponential growth has been observed in the last decade where enzymes were used as catalysts for polymerization of different monomers and due to enzyme's origin from natural sources they have been taken as a substitute for the metal based catalytic systems. Mild polymerization conditions, high enantio- and regio selectivity and recyclability of enzymes give them an extra edge over the use of organo-metallic catalysts. Though the enzyme-catalyzed polymerizations are environmentally highly advantageous, the high cost, large quantity of enzymes required for polymerization and formation of relatively low molecular weight polymers obstruct their employment in the industry. Due to these reasons, this technique is still at the stage of infancy to generate polymeric materials which can be converted to any useful physical form. In this article enzyme-catalyzed ring-opening polymerization of lactones, lactides, cyclic carbonates and depsipeptides has been reviewed with special focus on the molecular weight of the polymers synthesized hitherto using enzyme catalysis. It is necessary to obtain polymers of sufficient molecular weight from enzyme catalysis to withstand the specific requirements of their end applications if this technique is desired to be escalated to commercial level. PMID- 18406004 TI - Poly (amino acid) micelle nanocarriers in preclinical and clinical studies. AB - Polymeric micelles are expected to increase the accumulation of drugs in tumor tissues utilizing the EPR effect and to incorporate various kinds of drugs into the inner core by chemical conjugation or physical entrapment with relatively high stability. The size of the micelles can be controlled within the diameter range of 20 to 100 nm, to ensure that the micelles do not pass through normal vessel walls; therefore, a reduced incidence of the side effects of the drugs may be expected due to the decreased volume of distribution. These are several anticancer agent-incorporated micelle carrier systems under clinical evaluation. Phase 1 studies of a CDDP incorporated micelle, Nc-6004, and an sN-38 incorporated micelle, NK012, are now underway. A phase 2 study of a PTX incorporated micelle, NK105, against stomach cancer is also underway. PMID- 18406005 TI - The use of blind skin biopsy in the diagnosis of intravascular B-cell lymphoma. AB - Intravascular B-cell lymphoma is a rare type of non-Hodgkin's lymphoma that is characterized by a clonal proliferation of lymphoblasts within small blood vessels. Patients present with nonspecific symptoms and are often only given a diagnosis at autopsy. We report a case of intravascular B-cell lymphoma, characterized by pyrexia, anemia, thrombocytopenia, and mental status decline, without obvious cutaneous manifestations, that was diagnosed with blind skin biopsy. PMID- 18406006 TI - Perceived risks and perceived benefits of different nanotechnology foods and nanotechnology food packaging. AB - Nanotechnology has the potential to generate new food products and new food packaging. In a mail survey in the German speaking part of Switzerland, lay people's (N=337) perceptions of 19 nanotechnology applications were examined. The goal was to identify food applications that are more likely and food applications that are less likely to be accepted by the public. The psychometric paradigm was employed, and applications were described in short scenarios. Results suggest that affect and perceived control are important factors influencing risk and benefit perception. Nanotechnology food packaging was assessed as less problematic than nanotechnology foods. Analyses of individual data showed that the importance of naturalness in food products and trust were significant factors influencing the perceived risk and the perceived benefit of nanotechnology foods and nanotechnology food packaging. PMID- 18406007 TI - Underpaid women, stressed out men, satisfied emergency physicians. PMID- 18406008 TI - Neuropeptide interactions and REM sleep: a role for Urotensin II? AB - Urotensin II (UII) is a peptide with structural similarity to the somatostatin family with potent vasoconstrictor activity. UII receptor is expressed broadly in the periphery, and most notably in the heart and microvessels. In the brain, the UII receptor can be detected in the spinal cord and in cholinergic nuclei in the brainstem known to be involved in REM sleep regulation. Recent data suggest that, in addition to their vasoactive properties, UII receptor ligands may have excitatory activity on a selective group of neurons that modulate REM sleep. This review focuses on the implications of these findings for the neurobiology of REM sleep regulation and discusses the possible impact of UII and other neuropeptides on the balance of the alternation between sleep states. PMID- 18406009 TI - Neuropeptide FF receptors antagonist, RF9, attenuates opioid-evoked hypothermia in mice. AB - The present study used the endpoint of hypothermia to investigate opioid and neuropeptide FF (NPFF) interactions in conscious animals. Both opioid and NPFF systems played important roles in thermoregulation, which suggested a link between opioid receptors and NPFF receptors in the production of hypothermia. Therefore, we designed a study to investigate the relationship between opioid and NPFF in control of thermoregulation in mice. The selective NPFF receptors antagonist RF9 (30nmol) injected into the third ventricle failed to induce significant effect, but it completely antagonized the hypothermia of NPFF (45 nmol) after cerebral administration in mice. In addition, RF9 (30 nmol) co injected i.c.v. in the third ventricle reduced the hypothermia induced by morphine (5nmol,) or nociceptin/orphanin FQ (N/OFQ) (2 nmol). Neither the classical opioid receptors antagonist naloxone (10 nmol) nor NOP receptor antagonist [Nphe(1)]NC(1-13)NH(2) (7.5 nmol) reduced the hypothermia induced by the central injection of NPFF at dose of 45 nmol. Co-injected with a low dose of NPFF (5 nmol), the hypothermia of morphine (5 nmol) or N/OFQ (2 nmol) was not modified. These results suggest that NPFF receptors activation is required for opioid to produce hypothermia. In contrast, NPFF-induced hypothermia is mainly mediated by its own receptors, independent of opioid receptors in the mouse brain. This interaction, quantitated in the present study, is the first evidence that NPFF receptors mediate opioid-induced hypothermia in conscious animals. PMID- 18406010 TI - Zinc transporter 3 immunohistochemical tracing of sprouting mossy fibres. AB - Zinc transporter 3 (ZNT3) has been shown to transport zinc ions from the cytosol into presynaptic vesicles in the mammalian brain. Several studies have stated that the zinc ion containing synaptic vesicles of zinc-enriched neurons (ZEN) are loaded with ZNT3 proteins in their membranes. This fact makes it possible to trace sprouting mossy fibres in the temporal lobe epileptic hippocampus. In the present study, we examined the expression and distribution patterns of ZNT3 protein and chelatable zinc ions in the mouse hippocampus after pilocarpine treatment. Our results demonstrate that both ZNT3 immunostaining and autometallography reveal identical patterns of sprouting mossy fibres in the inner molecular layer in the mouse hippocampus. Using ZNT3 immuno-electron microscopic analysis we confirmed the presence of ectopic mossy fibre terminals in the inner molecular layer and found additionally by immuno-blotting a significant increase of ZNT3 in the pilocarpine-treated mouse hippocampi compared to age-matched controls. The increase of ZNT3 after pilocarpine treatment was time-dependent. The results support the notion that ZNT3 immunohistochemistry provides an excellent tool for tracing sprouting of ZEN terminals. The progressive increase of ZNT3 immunostaining in the temporal lobe epileptic hippocampus may relate to the increased levels of vesicular zinc ions during seizure. PMID- 18406012 TI - Antileukotrienic phenethylamido derivatives of arylalkanoic acids in the treatment of ulcerative colitis. AB - A series of arylalkanoic acid derivatives bearing methyl(phenethyl)amino groups were prepared and their inhibition of LTB(4) biosynthesis was evaluated. Regression analysis showed the slightly different parabolic dependences of this activity on lipophilicity of alpha-methyl and alpha-unsubstituted alkanoic acid derivatives. The relationship derived for alpha-unsubstituted alkanoic acids was extended by previously prepared group of similar derivatives of arylacetic acids without any change of regression coefficients and statistical criteria. It was concluded that the most active compounds belong to 2-arylpropanoic acid derivatives with lipophilicity close to logP(opt) (=6.97). But generally, the structural changes in the acidic part of compounds under study did not yield the substantial improvement of LTB(4) biosynthesis inhibition in comparison with the previously prepared series of derivatives IV. The anti-inflammatory effect of the compounds under study was evaluated in three animal models of inflammation and their possible utilization in the treatment of ulcerative colitis (UC) was followed. From 12 evaluated compounds, 4 compounds are more active in UC inhibition than the standard sulfasalazine but it can be stated that the change of connecting chain between aromatic ring and carboxyl did not bring about the important improvement of this activity in comparison with previous derivatives of arylacetic acids. Possible relation between LTB(4) biosynthesis inhibition and ulcerative colitis is seriously broken by the compound 8a with carbonyl as the additional functional group on the connecting chain between carboxyl and aromatic ring. PMID- 18406011 TI - Thiamine deficiency induces oxidative stress and exacerbates the plaque pathology in Alzheimer's mouse model. AB - Mitochondrial dysfunction, oxidative stress and reductions in thiamine-dependent enzymes have been implicated in multiple neurological disorders including Alzheimer's disease (AD). Experimental thiamine deficiency (TD) is an established model for reducing the activities of thiamine-dependent enzymes in brain. TD diminishes thiamine-dependent enzymes throughout the brain, but produces a time dependent selective neuronal loss, glial activation, inflammation, abnormalities in oxidative metabolism and clusters of degenerating neurites in only specific thalamic regions. The present studies tested how TD alters brain pathology in Tg19959 transgenic mice over expressing a double mutant form of the amyloid precursor protein (APP). TD exacerbated amyloid plaque pathology in transgenic mice and enlarged the area occupied by plaques in cortex, hippocampus and thalamus by 50%, 200% and 200%, respectively. TD increased Abeta(1-42) levels by about three fold, beta-CTF (C99) levels by 33% and beta-secretase (BACE1) protein levels by 43%. TD-induced inflammation in areas of plaque formation. Thus, the induction of mild impairment of oxidative metabolism, oxidative stress and inflammation induced by TD alters metabolism of APP and/or Abeta and promotes accumulation of plaques independent of neuron loss or neuritic clusters. PMID- 18406013 TI - Synthesis and evaluation of 4-amino-3,4-dihydro-2H-naphthalen-1-one derivatives as mast cell stabilising and anti-inflammatory compounds. AB - A novel series of amine and amide derivatives of 4-amino-3,4-dihydro-2H naphthalen-1-one were synthesised. The amine derivatives were evaluated for mast cell stabilising activity in rodent mast cell preparations against the reference compound disodium cromoglycate and found to possess significant activity in vitro. The amide compounds were evaluated in an in vivo murine model for anti inflammatory activity. PMID- 18406014 TI - Conformationally constrained analogues of N'-(4-tert-butylbenzyl)-N-(4 methylsulfonylaminobenzyl)thiourea as TRPV1 antagonists. AB - A series of bicyclic analogues having indan and tetrahydronaphthalene templates in the A-region were designed as conformationally constrained analogues of our previously reported potent TRPV1 antagonists (1, 3). The activities for rat TRPV1 of the conformationally restricted analogues were moderately or markedly diminished, particularly in the case of the tetrahydronaphthalene analogues. The analysis indicated that steric constraints at the benzylic position in the bicyclic analogues may be an important factor for their unfavorable interaction with the receptor. PMID- 18406015 TI - Pharmacophore modeling of diverse classes of p38 MAP kinase inhibitors. AB - Mitogen-activated protein (MAP) p38 kinase is a serine-threonine protein kinase and its inhibitors are useful in the treatment of inflammatory diseases. Pharmacophore models were developed using HypoGen program of Catalyst with diverse classes of p38 MAP kinase inhibitors. The best pharmacophore hypothesis (Hypo1) with hydrogen-bond acceptor (HBA), hydrophobic (HY), hydrogen-bond donor (HBD), and ring aromatic (RA) as features has correlation coefficient of 0.959, root mean square deviation (RMSD) of 1.069 and configuration cost of 14.536. The model was validated using test set containing 119 compounds and had high correlation coefficient of 0.851. The results demonstrate that results obtained in this study can be considered to be useful and reliable tools in identifying structurally diverse compounds with desired biological activity. PMID- 18406016 TI - Solution-phase parallel synthesis and evaluation of anticonvulsant activity of N substituted-3,4-dihydroisoquinoline-2(1H)-carboxamides. AB - In our previous studies we identified several isoquinoline derivatives displaying potent anticonvulsant effects in different animal models of epilepsy. With the aim to exploit the main structure-activity relationships (SAR) for this class of compounds we planned a solution-phase parallel synthesis (SPPS) of new N substituted-3,4-dihydroisoquinoline-2(1H)-carboxamides exploring the effect of introduction of different (cyclo)alkyl groups at carboxamide moiety linked to N-2 atom of isoquinoline scaffold. The pharmacological effects were evaluated against audiogenic seizures in DBA/2 mice and, even if some new derivatives were more active than valproate, the designed modifications did not improve the anticonvulsant efficacy with respect to their precursors. PMID- 18406017 TI - [Current concepts and treatment advances in systemic mastocytosis]. AB - PURPOSE: Mast cell disorders are defined by an abnormal accumulation of tissue mast cells in one or more organ systems. Clinical symptoms in mastocytosis result from mast cells derived mediators and, less frequently, from destructive infiltration of mast cells. Systemic mastocytosis is regressive among children, whereas the disease is persistent among adults. A clonal haematological non-mast cell lineage disease can be associated. The clinical course in these patients is variable ranging from asymptomatic for years to highly aggressive and rapidly devastating. Until recently, the only treatment of this incurable disease was symptomatic. CURRENT KNOWLEDGE AND KEY POINTS: Recent advances were done in understanding the physiopathology of this myeloproliferative syndrome which results from an activating mutation of the stem cell factor receptor: C-Kit. A somatic C-Kit mutation is usually detectable in mast cells and their progenitors. Different mutations were found and the mutation D816V is the most frequent. Their specific transduction paths were also studied. Diagnosis of systemic mastocytosis does not only rest upon pathological examination but also on molecular as well as immunological and immunochemical tools. FUTURE PROSPECTS AND PROJECTS: Physiopathological advancements led to suggest new treatments in order to directly inhibit proliferative paths of masts cells such as tyrosine kinase inhibitors and rapamycin. PMID- 18406018 TI - [Tropheryma whipplei and Whipple disease: false positive PCR detections of Tropheryma whipplei in diagnostic samples are rare]. AB - BACKGROUND: PCR can be used to detect T. whipplei (Tw) in samples from variable tissue types and body fluids. We report clinical, evolutive characteristics and final diagnosis in patients with positive Tw PCR assay. METHODS: Retrospective study of Tw PCR realized since 10years in a microbiology laboratory. RESULTS: Twenty-five Tw PCR assays were positive among 200 realized. Diagnosis was not confirmed in six cases. One patient was missing for follow up. Eighteen patients presented with Whipple's disease. Among these 18 patients, 14 had a classic Whipple's disease, three patients presented an endocarditis and one patient isolated neurological manifestations. Ten patients presented fever, seven a weight loss and 12 joint involvement. Four patients presented cutaneous manifestations, only six had gastrointestinal symptoms. Neurological involvement was reported in five cases, pulmonary symptoms in four cases, cardiac involvement in six cases and ocular signs in two cases. Anemia was reported in four patients and elevated levels of acute-phase reactants in 14 cases. Positive predictive value of Tw PCR for Whipple's disease diagnosis was 75%. Thirteen patients had a good evolution with antibiotics. Three patients presented recurrence and two cases with cardiovascular involvement died. CONCLUSION: Whipple's disease is rare but often mentioned in internist experience. The diagnosis should be every time confirmed. Tw PCR assay is an important diagnostic tool but is not sufficient to establish the diagnosis and must be interpreted with histopathology and immunohistochemical testing results. PMID- 18406019 TI - [Sciatica with motor loss revealing meningoradiculitis due to varicella-zoster virus]. AB - Herpes zoster is a disease which occurs secondary to the reactivation of varicella-zoster virus. Motor involvement in acute herpes zoster is rare. We report a case of sciatica L5 due to herpes zoster infection with motor loss. Typical skin lesions occurred one week before the sciatica. Radiological finding did not explain the paresis. The diagnosis of zoster sciatica with motor involvement was suspected. Serological tests and cerebrospinal fluid examination established the diagnosis. The antiviral and physical treatment was conducted in order to improve functional outcome. PMID- 18406020 TI - Expression of cytokine genes following pre- and post-hatch immunization of chickens with herpesvirus of turkeys. AB - Induction of immune response as characterised by expression of cytokine genes in the spleen following immunization of pre- and post-hatch chickens with herpesvirus of turkeys (HVT) vaccine was studied. The pattern of expression of IFN-gamma and IL-10 genes in pre-hatch immunized chickens was different from that observed in post-hatch HVT immunized chickens. This expression pattern of cytokine genes was associated with significantly higher HVT transcripts in pre hatch immunized chickens than in post-hatch immunized chickens. In conclusion, HVT immunization in chickens, irrespective of the age of immunization, stimulates host response characterised by the expression of cytokine genes, such as IFN gamma and IL-10 in the spleen. However, the age of immunization appears to influence the temporal pattern of IFN-gamma and IL-10 expression as well as replication of HVT. PMID- 18406021 TI - Bioneedles as vaccine carriers. AB - Bioneedles are small hollow mini implants fabricated from biodegradable polymers which can be filled with antigen. Bioneedles can be used for vaccination without syringes and needles. Formulations have been prepared containing tetanus toxoid with and without aluminum phosphate. Stability and immunogenicity of Bioneedles were compared with liquid formulations. The antigen, when formulated in Bioneedles, retained fully its antigenicity up to 60 degrees C for 1 week whereas the antigen, in its liquid form, lost all activity at 60 degrees C after 1 week. After 3 weeks at 60 degrees C, a recovery of 60% was still found in the Bioneedles. Mice injected with Bioneedles with adjuvanted tetanus toxoid showed a comparable functional antibody response to the group receiving conventional liquid injections. This response was achieved with a four times lower antigen concentration when using the Bioneedles compared to the regular injections. We conclude that Bioneedles are good alternatives for injections using needles and syringes. PMID- 18406022 TI - Derivation of transfer parameters for use within the ERICA Tool and the default concentration ratios for terrestrial biota. AB - An ability to predict radionuclide activity concentrations in biota is a requirement of any method assessing the exposure of biota to ionising radiation. Within the ERICA Tool fresh weight whole-body activity concentrations in organisms are estimated using concentration ratios (the ratio of the activity concentration in the organism to the activity concentration in an environmental media). This paper describes the methodology used to derive the default terrestrial ecosystem concentration ratio database available within the ERICA Tool and provides details of the provenance of each value for terrestrial reference organisms. As the ERICA Tool considers 13 terrestrial reference organisms and the radioisotopes of 31 elements, a total of 403 concentration ratios were required for terrestrial reference organisms. Of these, 129 could be derived from literature review. The approaches taken for selecting the remaining values are described. These included, for example, assuming values for similar reference organisms and/or biogeochemically similar elements, and various simple modelling approaches. PMID- 18406023 TI - Cystic changes in dental follicle associated with radiographically normal impacted mandibular third molar. AB - The aim of the study was to find out the incidence of histological abnormalities in soft tissues that are associated with impacted lower third molars with no pericoronal lytic lesion. We studied 100 patients who had a radiographically normal asymptomatic impacted lower third molar (<2.4mm radiolucency). The incidence of cystic changes in the dental follicle was 46%. There was a high incidence in the right side and in men. We recommend histopathological evaluation and radiographic diagnosis in the management of impacted third molars. PMID- 18406024 TI - Modelling the spatial distribution of ammonia emissions in the UK. AB - Ammonia emissions (NH3) are characterised by a high spatial variability at a local scale. When modelling the spatial distribution of NH3 emissions, it is important to provide robust emission estimates, since the model output is used to assess potential environmental impacts, e.g. exceedance of critical loads. The aim of this study was to provide a new, updated spatial NH3 emission inventory for the UK for the year 2000, based on an improved modelling approach and the use of updated input datasets. The AENEID model distributes NH3 emissions from a range of agricultural activities, such as grazing and housing of livestock, storage and spreading of manures, and fertilizer application, at a 1-km grid resolution over the most suitable landcover types. The results of the emission calculation for the year 2000 are analysed and the methodology is compared with a previous spatial emission inventory for 1996. PMID- 18406025 TI - The amphipod Hyalella azteca as a biomonitor in field deployment studies for metal mining. AB - Specimens of the amphipod Hyalella azteca were deployed, in June-July 2003, along metal contamination gradients in two rivers affected by metal mining in the Abitibi-James Bay region, northwestern Quebec. The amphipods were placed along with natural food items in small, acrylic cages and left in six riverine sites for 17 days. Twelve metals (As, Cu, La, Mn, Ni, Sb, Se, Tl, U, V, Zn, and CrO4(2 ) modelled by WHAM VI) in transplanted H. azteca varied along metal contamination gradients in a consistent manner, i.e., as a function of metal exposure. Bioaccumulation of As, Cr, La, Ni, Sb, Se, Tl, U and V, as defined by a field BCF, was significantly correlated with their chronic toxicity potential towards the amphipod. We conclude that H. azteca may be a useful field biomonitor for metal mining. In addition, our results suggest that such biomonitoring programs should include less studied elements such as Se in mining effluents. PMID- 18406026 TI - Sorption of polycyclic aromatic hydrocarbons (PAHs) to carbonaceous materials in a river floodplain soil. AB - We report on sorption isotherm of phenanthrene (Phe) for river floodplain soil associated with carbonaceous materials, with particular attention being devoted to the natural loading of Phe. Our sorption experiments with original soil samples, size, and density sub-fractions showed that the light fraction had the highest sorption capacity comparable to low rank coals. In addition, the light fraction contributed most for the sorption of Phe in total soil samples. K(oc) values for all fractions were in the same range, thus indicating that coal and coal-derived particles in all samples are responsible for the enhanced sorption for Phe. Sorption was strongly nonlinear and the combined partitioning and pore filling model gave a better fit than the Freundlich sorption model. In addition, the spiked PAHs did not show the same behavior as the naturally aged ones, therefore the accessibility of indigenous background organic contaminants was reduced when coal and coal-derived particles are associated with the soils. PMID- 18406027 TI - The independent predictive value of peritraumatic dissociation for PTSD symptomatology after type I trauma: a systematic review of prospective studies. AB - We conducted a systematic review of prospective studies examining the independent predictive value of peritraumatic dissociation (PD) for posttraumatic stress disorder (PTSD) symptomatology following single traumatic events. Insight into the independent predictive value may help to identify victims at risk for PTSD symptomatology. For this purpose a literature search was carried out using the online databases PsycINFO, Medline/Pubmed, and PILOTS. Studies were included if they were published in peer-reviewed journals (before 2007), focused on more or less single traumatic events (Type I trauma), assessed PD within 1 month, had follow-ups 3 months or later, and which controlled for mental health problems at the time PD was assessed. The majority of the 17 identified studies showed no or only weak indications of an independent predictive value of PD for PTSD symptomatology following type I traumas. Only 3 of the 6 studies with positive results reported a strong independent predictive value of PD. Although bivariately associated with PTSD symptomatology, there is no general consensus across prospective Type I trauma studies that PD qualifies as an important independent predictor of PTSD symptomatology. Results indicate that initial mental health problems, among other factors, are better predictors of PTSD symptomatology than PD. PMID- 18406028 TI - Dermal sensitization quantitative risk assessment (QRA) for fragrance ingredients. AB - Based on chemical, cellular, and molecular understanding of dermal sensitization, an exposure-based quantitative risk assessment (QRA) can be conducted to determine safe use levels of fragrance ingredients in different consumer product types. The key steps are: (1) determination of benchmarks (no expected sensitization induction level (NESIL)); (2) application of sensitization assessment factors (SAF); and (3) consumer exposure (CEL) calculation through product use. Using these parameters, an acceptable exposure level (AEL) can be calculated and compared with the CEL. The ratio of AEL to CEL must be favorable to support safe use of the potential skin sensitizer. This ratio must be calculated for the fragrance ingredient in each product type. Based on the Research Institute for Fragrance Materials, Inc. (RIFM) Expert Panel's recommendation, RIFM and the International Fragrance Association (IFRA) have adopted the dermal sensitization QRA approach described in this review for fragrance ingredients identified as potential dermal sensitizers. This now forms the fragrance industry's core strategy for primary prevention of dermal sensitization to these materials in consumer products. This methodology is used to determine global fragrance industry product management practices (IFRA Standards) for fragrance ingredients that are potential dermal sensitizers. This paper describes the principles of the recommended approach, provides detailed review of all the information used in the dermal sensitization QRA approach for fragrance ingredients and presents key conclusions for its use now and refinement in the future. PMID- 18406029 TI - Risk assessments for the insect repellents DEET and picaridin. AB - For the use of topical insect repellents, DEET and picaridin, human health risk assessments were conducted for various population subgroups. Acute, subchronic, and chronic dermal exposures were examined. No-observed-effect-levels (NOELs) of 200, 300, and 100mg/kg body weight (BW) were used as endpoints for DEET for acute, subchronic, and chronic exposures, respectively. For picaridin, a NOEL of 2000 mg/kg BW/day for acute exposure and a NOEL of 200 mg/kg BW/day for subchronic and chronic exposures were used. Daily exposures to several population subgroups were estimated. Risks were characterized using the Margin of Exposure (MOE) method (NOEL divided by the estimated exposure), whereby estimated MOEs were compared to an MOE of 100. Estimates of daily exposures ranged from 2 to 59 mg/kg BW/day for DEET and 2 to 22 mg/kg BW/day for picaridin. Children had the lowest MOEs. However, none of the estimated exposures exceeded NOELs for either repellent. At 40% DEET for acute exposure, children < or = 12 years had MOEs below 100. For subchronic and chronic exposures children at > or = 25% DEET and at 15% picaridin had MOEs below 100. Therefore, we found no significant toxicological risks from typical usage of these topical insect repellents. PMID- 18406030 TI - Who's talking? Communication between health providers and HIV-infected adults related to herbal medicine for AIDS treatment in western Uganda. AB - Communication between patients and physicians about herbal medicine is valuable, enabling physicians to address issues of potential herb-drug interactions and ensuring appropriate medical care. As seemingly harmless herbal remedies may have detrimental interactions with various HIV antiretroviral drugs, the importance of communication is intensified, but often stifled around the use of herbal medicine in the treatment of HIV/AIDS. In western Uganda, 137 HIV-infected adults attending conventional HIV/AIDS treatment programmes (67 of whom were receiving antiretroviral therapy) shared their experiences and perceptions about traditional herbal medicine and related patient-physician communication issues through interviews and focus group discussions. Although close to 64% of respondents reported using herbal medicine after being diagnosed with HIV, only 16% of these respondents had informed their conventional medical practitioners about using these herbs. Furthermore, only 13% of antiretroviral therapy recipients had inquired about concurrent herb-antiretroviral drug use with their HIV/AIDS treatment providers, largely because they perceived a low acceptance and support for herbal medicine by conventional medical practitioners. Importantly however, almost 68% of HIV-infected adults indicated they would be willing to discuss herbal medicine use if directly asked by a conventional medical practitioner, and the overwhelming majority (91%) said they were amenable to following physician advice about herbal medicine. As such, improved patient physician communication about herbal medicine is needed, and we recommend that herbal medicine histories be completed when patient histories are taken. Also, HIV/AIDS treatment programmes should be encouraged to develop specific patient physician communication standards and best practice guidelines to ensure that patients can make informed decisions about herb and pharmaceutical drug co therapy based on known risks, particularly in the case of AIDS patients receiving antiretroviral therapy. Communication about herbal medicine usage should be viewed as a timely and cost-effective component of antiretroviral therapy programmes, one which may contribute to the overall success of AIDS treatment in Africa. PMID- 18406031 TI - Are 'anonymous' and 'non-directed' prerequisites for living altruistic donation? The views of transplant physicians from France and Quebec. AB - It can be argued that living altruistic donors should remain anonymous and should not express preferences in the selection of organ recipients. This study aimed to describe the views of transplant physicians in France and Quebec regarding these issues. A total of 27 French and 19 Quebec renal transplant physicians took part in individual, semi-directed interviews. Almost all of the physicians agreed that anonymity is mandatory in living altruistic donation (LAD). Regarding the issue of directed donation, most of the French physicians (78%) were opposed to any form of the practice, compared to only a third of their Quebec colleagues (32%). We found that these positions were embedded in their respective cultural, legal and social contexts. These results afford a better understanding of these complex issues in two different cultural contexts, and will be useful in the development of international guidelines for LAD. PMID- 18406032 TI - Acetylcholine esterase activity and behavioral response in hypoxia induced neonatal rats: effect of glucose, oxygen and epinephrine supplementation. AB - Brain damage due to an episode of hypoxia remains a major problem in infants causing deficit in motor and sensory function. Hypoxia leads to neuronal functional failure, cerebral palsy and neuro-developmental delay with characteristic biochemical and molecular alterations resulting in permanent or transitory neurological sequelae or even death. During neonatal hypoxia, traditional resuscitation practices include the routine administration of 100% oxygen, epinephrine and glucose. In the present study, we assessed the changes in the cholinergic system by measuring the acetylcholinesterase (AChE) activity and the behavioral responses shown by hypoxia induced neonatal rats and hypoxic rats supplemented with glucose, oxygen and epinephrine using elevated plus-maze and open-field test. The acetylcholine esterase enzyme activity showed a significant decrease in cerebral cortex, whereas it increased significantly in the muscle of experimental rats when compared to control. Hypoxic rats supplemented with glucose, glucose and oxygen showed a reversal to the control status. Behavioral studies were carried out in experimental rats with elevated plus-maze test and open-field test. Hypolocomotion and anxiogenic behavioral responses were observed in all experimental rats when compared to control, hypoxic rats supplemented with glucose, glucose and oxygen. Thus, our results suggest that brain damage due to hypoxia, oxygen and epinephrine supplementation in the neonatal rats cause acetylcholine-neuromuscular-defect leading to hypolocomotion and anxiogenic behavioral response. Glucose and glucose with oxygen supplementation to hypoxic neonates protect the brain damage for a better functional status in the later life. PMID- 18406033 TI - Letter by Namazi regarding article "Simvastatin attenuates cisplatin-induced kidney and liver damage in rats". PMID- 18406034 TI - Routine oral examinations in primary care: which predictors determine what is done? A prospective clinical case recording study. AB - OBJECTIVES: Elements of a routine oral examination (ROE) in dental practice may be determined by patients' oral status, as recommended by prevailing knowledge, as well as by other factors. Our aim was to identify patient and GDP characteristics associated with aspects of oral health assessment (OHA) and clinical management (CM) in patients with various oral conditions. METHODS: A prospective observational study was performed, based on clinical case recording of 1059 ROEs by 128 GDPs. A clinical case recording form was used to record oral health assessment, diagnoses made, and clinical management for each ROE. Multilevel logistic regression analyses (with random coefficients) were performed. RESULTS: Overall, 'patients' age' in domains OHA as well as CM was the most salient predictor, while 'positive attitude to periodontal screening' showed to be a prominent GDP-factor. Patient characteristics mostly involved in OHA and CM were 'disease-free period' (odds ratios from 0.21 to 0.66), 'oral health compliance' (odds ratios from 0.32 to 0.65) and 'risk for periodontal disease' (odds ratios from 1.79 to 4.97). 'Continuing professional development' (odds ratios from 2.54 to 4.95), 'mean reading hours' (odds ratios from 2.25 to 4.48) and 'cooperation with peers' (odds ratios from 2.78 to 3.72) showed to be significant GDP-predictors. CONCLUSIONS: ROEs are determined by patient oral health status, particularly by aspects of oral health compliance and risk for oral disease, but also by GDP characteristics. The latter may reflect perceptions of a professional role, which need to be considered in efforts to improve the quality of ROE in oral care. PMID- 18406035 TI - Dynamics and state transitions during resuscitation in out-of-hospital cardiac arrest. AB - BACKGROUND: The state or rhythm during resuscitation, i.e. ventricular fibrillation/tachycardia (VF/VT), asystole (ASY), pulseless electrical activity (PEA), or return of spontaneous circulation (ROSC) determines management. The state is unstable and will change either spontaneously (e.g. PEA-->ASY) or by intervention (e.g. VF-->ASY after DC shock); temporary ROSC may also occur. To gain insight into the dynamics of this process, we analyzed the state transitions over time using real-life data. METHODS: Detailed recordings from 304 episodes of attempted resuscitation from out-of-hospital cardiac arrests of presumed cardiac etiology were obtained from modified Heartstart 4000 defibrillators. State transitions were visualized and described, and analyzed in terms of a Markov probability model. RESULTS: The median number of state transitions was 5 (range 1 39), and more transitions were observed with VF than PEA or asystole as the initial rhythm. Of 105 patients (35%) who regained ROSC at some point during CPR, only 65 (21%) achieved sustained ROSC; suggesting an unrealized survival potential. A 3-min transition probability matrix was estimated: for example, a patient early in VF has a probability of 31% to be in ASY, 32% of still being in VF, 5% to have temporary ROSC, and 2% to have sustained ROSC after 3 min. CONCLUSION: The dynamics of resuscitation can be described in terms of state transitions and a Markov probability model. This framework enables prediction of short-term clinical development, supports informed decisions during CPR, and suggests a novel area for research. PMID- 18406037 TI - Evaluation of staff's retention of ACLS and BLS skills. AB - OBJECTIVES/PURPOSE: To test registered nurses' abilities to retain basic or advanced life support psychomotor skills and theoretical knowledge. DESIGN: A repeated-measures, quasi-experimental design was used. METHODS: Written and performance tests (initial, post-training, and final testing) used scenarios requiring performance of advanced cardiac life support (ACLS) or basic life support (BLS) skills. Final testing was by random assignment to 3, 6, 9, or 12 months. SAMPLE: A convenience sample (n=133) was used. INSTRUMENTATION: American Heart Association 2000 ACLS and BLS evaluation tools were used in a simulated testing environment. FINDINGS: Findings show nurses retain theoretical knowledge but performance skills degrade quickly. ACLS skills degrade faster than BLS skills with 63% passing BLS at 3 months and 58% at 12 months. Only 30% of participants passed ACLS skills at 3 months and 14% at 12 months. These findings are similar to the results of other investigators in over a decade of research. CONCLUSIONS: Study results showed a decline in skills retention with nurses unable to perform ACLS and BLS skills to standard for the entire certification period. The need for more frequent refresher training is needed. No formal research at this institution indicates skill degradation adversely affected patient outcomes. Further research on ACLS and BLS course content, design, management, and execution is needed. PMID- 18406036 TI - Liquid ventilation with perfluorocarbons facilitates resumption of spontaneous circulation in a swine cardiac arrest model. AB - BACKGROUND: Induced external hypothermia during ventricular fibrillation (VF) improves resuscitation outcomes. Our objectives were twofold (1) to determine if very rapid hypothermia could be achieved by intrapulmonary administration of cold perfluorocarbons (PFC), thereby using the lungs as a vehicle for targeted cardiopulmonary hypothermia, and (2) to determine if this improved resuscitation success. METHODS: Part 1: Nine female swine underwent static intrapulmonary instillation of cold perfluorocarbons (PFC) during electrically induced VF. Part 2: Thirty-three female swine in VF were immediately ventilated via total liquid ventilation (TLV) with pre-oxygenated cold PFC (-15 degrees C) or warm PFC (33 degrees C), while control swine received no ventilation during VF. All swine in both Parts 1 and 2 underwent VF arrest for 11 min, then defibrillation, ventilation and closed chest massage until resumption of spontaneous circulation (ROSC). The endpoint was continued spontaneous circulation for 1h without pharmacologic support. RESULTS: Static intrapulmonary instillation of cold PFC achieved rapid cardiopulmonary hypothermia; pulmonary artery (PA) temperature of 33.5+/-0.2 degrees C was achieved by 10 min. Nine of 9 achieved ROSC. Hypothermia was achieved faster using TLV: at 6 min VF, cold TLV temperature was 32.9+/-0.4 degrees C vs. cold static instillation temperature 34.3+/-0.2 degrees C. Nine of 11 cold TLV swine achieved ROSC for 1h vs. 3 of 11 control swine (p=0.03). Warm PFC also appeared to be beneficial, with a trend toward greater achievement of ROSC than control (ROSC; warm PFC 8 of 11 vs. control 3 of 11, p=0.09). CONCLUSION: Targeted cardiopulmonary intra-arrest moderate hypothermia was achieved rapidly by static intrapulmonary administration of cold PFC and more rapidly by total liquid ventilation with cold PFC; resumption of spontaneous circulation was facilitated. Warm PFC showed a trend toward facilitating ROSC. PMID- 18406038 TI - Derivation and validation of a score based on Hypotension, Oxygen saturation, low Temperature, ECG changes and Loss of independence (HOTEL) that predicts early mortality between 15 min and 24 h after admission to an acute medical unit. AB - BACKGROUND: Predictive scores such as APACHE II have been used to assess patients in intensive care units, but few scores have been used to assess acutely ill general medical patients. DESIGN: Examination of the ability of clinical variables documented at the time of admission to predict early mortality between 15 min and 24 h after admission. SETTING: An Irish rural hospital. SUBJECTS: 10,290 consecutive patients admitted as acute medical emergencies, divided into a derivation cohort of 6947 patients and a validation cohort of 3343 patients. RESULTS: 40 patients of the derivation cohort (0.6%) died within 24h of hospital admission. Multivariate analysis revealed 11 independent predictors of early death from which a simplified model with minimal loss of predictive ability was derived. Since this model contained only the five variables of Hypotension (systolic blood pressure<100 mm Hg), low Oxygen saturation (<90%), low Temperature (<35 degrees C, abnormal ECG and Loss of independence (unable to stand unaided) it was named the HOTEL score (one point for each variable). There were no differences in the early mortality predicted by this score between the derivation and validation cohorts-the area under the receiver operator characteristic curves for the derivation and validation cohorts were 86.5% and 85.4%, respectively. None of the patients with a score of zero died within 15 min and 24 h and a score of one had an early mortality of 0.3% in both cohorts. A score of two had an early mortality of 0.9% in the derivation cohort and 1.7% in the validation cohort, while a score of three or greater had an early mortality of 10.2% in the derivation and 5.6% the validation cohort. CONCLUSIONS: The HOTEL score quickly identifies patients at a low and high risk of death between 15 min and 24 h after admission, thus enabling prompt triage and placement within a health care facility. PMID- 18406039 TI - Anti-arrhythmic and vasopressor medications for the treatment of ventricular fibrillation in severe hypothermia: a systematic review of the literature. AB - OBJECTIVE: To determine the rate of return of spontaneous circulation (ROSC) in animal models performing resuscitation from induced ventricular fibrillation (VF) in severe hypothermia (<30 degrees C). MATERIALS AND METHODS: A medical literature database search from 1966 to present was performed identifying placebo controlled trials using anti-arrhythmic or vasopressor medications to treat ventricular fibrillation in the setting of severe hypothermia. RESULTS: 7 controlled studies were identified (n=117) testing 6 combinations of resuscitative medications. ROSC rates for treatment versus control groups were as follows: amiodarone (6% vs. 18%, p=0.6, n=34), bretylium (35% vs. 35%, p=1.0, n=40), intermediate- and high-dose epinephrine (adrenaline) (36% vs. 27%, p=1.0, n=22), vasopressin (60% vs. 0%, p<0.0001, n=39), vasopressin and amiodarone (0% vs. 0%, p=NS, n=11), low-dose epinephrine and amiodarone (91% vs. 30%, p=0.0075, n=21). Cumulatively, among all studies administering vasopressors, the rate of ROSC was 62% in treatment groups contrasted to 17% in control groups (p<0.0001, n=77). CONCLUSIONS: In controlled animal models of severe hypothermia, ROSC rates for induced ventricular fibrillation are higher with utilization of vasopressor medications. Current guidelines which recommend withholding these medications in the setting of hypothermic cardiac arrest should be re-evaluated. PMID- 18406040 TI - Elevating blood glucose level increases the retention of information from a public safety video. AB - Seventy-two undergraduates, 41 female and 31 male, aged 19-38, who had eaten breakfast had their blood glucose levels measured at 10.00 a.m. Following this they watched a presentation on safety when living away from home and had their recall of this material tested. Then half the participants imbibed a drink containing 50 g of glucose while the other participants imbibed a drink containing saccharine. A 20 min lecture followed to allow absorption and then blood glucose level was again measured and a second safety presentation was followed by another memory test. The two groups did not differ in initial blood glucose levels but only the group drinking the glucose drink had significantly elevated blood glucose at second test. The two groups did not differ in memory recall at the first test but the glucose drink group recalled approximately 20% more material after the drink. Analysis of covariance on the memory data with post-drink blood glucose level as a covariate attenuated the glucose effect suggesting that there is a relationship between glucose level and memory performance on this task. It is concluded that, although the precise mechanism by which enhanced memory performance occurs cannot be elucidated by these data, the results have practical significance for education. It is suggested that breakfast consisting of low glycaemic index foods may aid in challenging learning situations. PMID- 18406041 TI - Chemoprevention of human prostate cancer by green tea catechins: two years later. A follow-up update. PMID- 18406042 TI - Digital rectal examination remains an important screening tool for prostate cancer. PMID- 18406043 TI - Editorial comment on: TGF-beta1 Inhibits Cx43 expression and formation of functional syncytia in cultured smooth muscle cells from human detrusor. PMID- 18406045 TI - Digital rectal examination and the diagnosis of prostate cancer--a study based on 8 years and three screenings within the European Randomized Study of Screening for Prostate Cancer (ERSPC), Rotterdam. AB - BACKGROUND: Evidence indicates that an abnormal digital rectal examination (DRE) is a risk factor for high-grade prostate cancer (PC). OBJECTIVE: To determine whether men with an initially suspicious DRE, a prostate-specific antigen (PSA) level > or = 3.0 ng/ml, and a benign prostate biopsy are at higher risk for significant PC at rescreening than men with an initially normal DRE, and whether an adaptation of the rescreening interval is warranted for this group. DESIGN, SETTING, AND PARTICIPANTS: Within the European Randomized Study of Screening for Prostate Cancer (ERSPC), Rotterdam, 2218 men underwent biopsy of the prostate (from 1993 to 2000) with a benign result at initial screening. The serum PSA was determined every 4 yr. A PSA level of > or = 3.0 ng/ml prompted a DRE and a lateralised sextant biopsy. MEASUREMENTS: Number and characteristics of PCs found at repeat screenings and as interval cancers (ICs) were compared between men with or without a suspicious DRE result at initial screening. Multivariate logistic regression analyses were performed to evaluate if an initially suspicious DRE was a significant predictor for detecting cancer at consecutive screenings. RESULTS AND LIMITATIONS: After 4 yr, the total number of PCs detected in men with and without an initially suspicious DRE was, respectively, 27 (6%) versus 103 (6%) (p=0.99). After 8 yr these numbers increased, respectively, to 45 (10%) versus 167 (10%) (p=0.88). The proportion of clinically significant PCs was 2% and 3%, respectively, for the group with initially normal and abnormal DRE after 8 yr. Having a suspicious DRE result at initial screening was not a significant predictor for detecting PC after 4 yr [odds ratio (OR)=1.15, p=0.59) or 8 yr (OR=1.41, p=0.43)]. A limitation of this study is the relatively short follow-up of 8 yr. CONCLUSIONS: During a follow-up of 8 yr after initial cancer-negative biopsy, an initially suspicious DRE did not influence the chance for detection of cancer or significant cancer at later screens. An adaptation of the rescreening interval on the basis of the initial DRE-outcome is not warranted in future population-based screening for prostate cancer. PMID- 18406046 TI - Editorial comment on: Digital rectal examination and the diagnosis of prostate cancer--a study based on 8 years and three screenings within the European Randomized Study of Screening for Prostate Cancer (ERSPC), Rotterdam. PMID- 18406047 TI - The management of category T1a-T1b (incidental) prostate cancer: can we predict who needs treatment? PMID- 18406048 TI - TGF-beta1 inhibits Cx43 expression and formation of functional syncytia in cultured smooth muscle cells from human detrusor. AB - BACKGROUND: Human detrusor smooth muscle cells (hBSMCs) are coupled by connexin 43 (Cx43)-positive gap junctions to form functional syncytia. Gap junctional communication likely is necessary for synchronised detrusor contractions and is supposed to be altered in voiding disturbances. Other authors have shown that the pleiotropic cytokine TGF-beta1 upregulates Cx43 expression in human aortic smooth muscle cells. OBJECTIVE: In this study, we examined the TGF-beta1 effects on Cx43 expression in cultured hBSMCs. DESIGN, SETTING, AND PARTICIPANTS: hBSMC cultures, established from patients undergoing cystectomy, were treated with recombinant human TGF-beta1. MEASUREMENTS: Cx43 expression was then examined by Western blotting, real-time PCR, and immunocytochemistry. Dye-injection experiments were used to study the size of functional syncytia. RESULTS AND LIMITATIONS: Dye coupling experiments revealed stable formation of functional syncytia in passaged cell cultures (P1-P4). Stimulation with TGF-beta1 led to significant reduction of Cx43 immunoreactivity and coupling. Cx43 protein expression was significantly downregulated and Cx43 mRNA was only 30% of the control level. Interestingly, low phosphorylation species of Cx43 were particularly affected. CONCLUSIONS: Our experiments demonstrated a significant down regulation of connexin 43 by TGF beta1 in cultured hBSMCs. These findings support the view that TGF-beta1 is involved in the pathophysiology of urinary bladder dysfunction. PMID- 18406049 TI - New insight into the molecular evolution of the somatostatin family. AB - The present review describes the molecular evolution of the somatostatin family and its relationships with that of the urotensin II family. Most of the somatostatin sequences collected from different vertebrate species can be grouped as the products of at least four loci. The somatostatin 1 (SS1) gene is present in all vertebrate classes from agnathans to mammals. The SS1 gene has given rise to the somatostatin 2 (SS2) gene by a segment/chromosome duplication that is probably the result of a tetraploidization event according to the 2R hypothesis. The somatostatin-related peptide cortistatin, first identified in rodents and human, is the counterpart of SS2 in placental mammals. In fish, the existence of two additional somatostatin genes has been reported. The first gene, which encodes a peptide usually named somatostatin II (SSII), exists in almost all teleost species investigated so far and is thought to have arisen through local duplication of the SS1 gene. The second gene, which has been characterized in only a few teleost species, encodes a peptide also named SSII that exhibits a totally atypical structure. The origin of this gene is currently unknown. Nevertheless, because the two latter genes are clearly paralogous genes, we propose to rename them SS3 and SS4, respectively, in order to clarify the current confusing nomenclature. The urotensin II family consists of two genes, namely the urotensin II (UII) gene and the UII-related peptide (URP) gene. Both UII and URP exhibit limited structural identity to somatostatin so that UII was originally described as a "somatostatin-like peptide". Recent comparative genomics studies have revealed that the SS1 and URP genes, on the one hand, and the SS2 and UII genes, on the other hand, are closely linked on the same chromosomes, thus confirming that the SS1/SS2 and the UII/URP genes belong to the same superfamily. According to these data, it appears that an ancestral somatostatin/urotensin II gene gave rise by local duplication to a somatostatin ancestor and a urotensin II ancestor, whereupon this pair was duplicated (presumably by a segment/chromosome duplication) to give rise to the SS1-UII pair and the SS2-URP pair. PMID- 18406050 TI - Endocannabinoids, sperm functions and energy metabolism. AB - Cannabinoids, the main active components of marijuana, have been shown to exert different adverse effects on male reproduction both in vertebrates and invertebrates. The main effects of endocannabinoids, a particular group of endogenously produced cannabinoids, in sperm are the inhibition of motility, capacitation and acrosome reaction, all fundamental processes necessary for oocyte penetration, whose alteration leads to the inhibition of sperm fertilizing ability. These inhibitory effects are mediated by the direct action of endocannabinoids on sperm through the activation of the cannabinoid receptor subtype 1 that has been shown to be expressed in mature sperm. In many different cell types it has been demonstrated that endocannabinoids negatively influence mitochondrial activity. In the present paper it will be briefly reviewed the role of endocannabinoids, on sperm motility, capacitation and acrosome reaction with particular attention on the possible interference of endocannabinoids with sperm mitochondrial activity. PMID- 18406053 TI - Combined Fenton oxidation and aerobic biological processes for treating a surfactant wastewater containing abundant sulfate. AB - The present study is to investigate the treatment of a surfactant wastewater containing abundant sulfate by Fenton oxidation and aerobic biological processes. The operating conditions have been optimized. Working at an initial pH value of 8, a Fe2+ dosage of 600mgL(-1) and a H2O2 dosage of 120mgL(-1), the chemical oxidation demand (COD) and linear alkylbenzene sulfonate (LAS) were decreased from 1500 and 490mgL(-1) to 230 and 23mgL(-1) after 40min of Fenton oxidation, respectively. Advanced oxidation pretreatment using Fenton reagent was very effective at enhancing the biodegradability of this kind of wastewater. The wastewater was further treated by a bio-chemical treatment process based on an immobilized biomass reactor with a hydraulic detention time (HRT) of 20h after Fenton oxidation pretreatment under the optimal operating conditions. It was found that the COD and LAS of the final effluent were less than 100 and 5mgL(-1), corresponding to a removal efficiencies of over 94% and 99%, respectively. PMID- 18406054 TI - Chromium (III) removal by weak acid exchanger Amberlite IRC-50 (Na). AB - Chromium (III) sorption on a weakly macroporous cation exchanger Amberlite IRC-50 (Na) is studied as a function of time and temperature. The rate constant values for chromium (III) sorption are calculated both for film and particle diffusion processes. The temperature is found to have a positive effect on both the diffusional processes. The high values of energy of activation confirm the particle diffusional nature of the process. The pH changes in the system show a competition between the Cr(OH)(2+) and H(+) ions for the exchange sites of the adsorbent. Equilibrium data are explained with the help of Langmuir equation. Various thermodynamic parameters (DeltaH degrees , DeltaS degrees and DeltaG degrees) from chromium (III) exchange on the resin are calculated. PMID- 18406051 TI - Reactive oxygen species and angiogenesis: NADPH oxidase as target for cancer therapy. AB - Angiogenesis is essential for tumor growth, metastasis, arteriosclerosis as well as embryonic development and wound healing. Its process is dependent on cell proliferation, migration and capillary tube formation in endothelia cells (ECs). High levels of reactive oxygen species (ROS) such as superoxide and H2O2 are observed in various cancer cells. Accumulating evidence suggests that ROS function as signaling molecules to mediate various growth-related responses including angiogenesis. ROS-dependent angiogenesis can be regulated by endogenous antioxidant enzymes such as SOD and thioredoxin. Vascular endothelial growth factor (VEGF), one of the major angiogenesis factor, is induced in growing tumors and stimulates EC proliferation and migration primarily through the VEGF receptor type2 (VEGFR2, Flk1/KDR). Major source of ROS in ECs is a NADPH oxidase which consists of Nox1, Nox2, Nox4, Nox5, p22phox, p47phox and the small G-protein Rac1. NADPH oxidase is activated by various growth factors including VEGF and angiopoietin-1 as well as hypoxia and ischemia, and ROS derived from this oxidase are involved in VEGFR2 autophosphorylation, and diverse redox signaling pathways leading to induction of transcription factors and genes involved in angiogenesis. Dietary antioxidants appear to be effective for treatment of tumor angiogenesis. The aim of this review is to provide an overview of the recent progress on role of ROS derived from NADPH oxidase and redox signaling events involved in angiogenesis. Understanding these mechanisms may provide insight into the NADPH oxidase and redox signaling components as potential therapeutic targets for tumor angiogenesis. PMID- 18406055 TI - Magnetically separable composite photocatalyst with enhanced photocatalytic activity. AB - A novel magnetically separable composite photocatalyst, anatase titania-coated magnetic activated carbon (TMAC), was prepared in this article. In the synthesis, magnetic activated carbon (MAC) was firstly obtained by adsorbing magnetic Fe3O4 nanoparticles onto the activated carbon (AC), and then the obtained MAC was directly coated by anatase titania nanoparticles prepared at low temperature (i.e. 75 degrees C). The prepared samples were characterized by XRD, SEM and vibrating sample magnetometer (VSM). The composite photocatalyst can be easily separated from solution by a magnet, its photocatalytic activity in degradation of phenol in aqueous solution also has dramatic enhancement compared to that of the neat titania. PMID- 18406052 TI - Cortactin in tumor invasiveness. AB - Cortactin is a cytoskeletal protein and src kinase substrate that is frequently overexpressed in cancer. Animal studies suggest that cortactin overexpression increases tumor aggressiveness, possibly through promotion of tumor invasion and metastasis. Recently, many studies have documented a role for cortactin in promoting cell motility and invasion, including a critical role in invadopodia, actin rich-subcellular protrusions associated with degradation of the extracellular matrix by cancer cells. Here, I review the evidence and potential mechanisms for cortactin as a critical mediator of tumor cell invasion. PMID- 18406056 TI - Removal of gallium (III) ions from acidic aqueous solution by supercritical carbon dioxide extraction in the green separation process. AB - Supercritical carbon dioxide extraction, which is a feasible "green" alternative, was applied in this study as a sample pretreatment step for the removal of gallium (III) ions from acidic aqueous solution. The effect of various process parameters, including various chelating agents, extraction pressure and temperature, dimensionless CO(2) volume, the concentration of the chelating agent, and the pH of the solution, governing the efficiency and throughput of the procedure were systematically investigated. The performance of the various chelating agents from different studies indicated that the extraction efficiency of supercritical CO(2) was in the order: thiopyridine (PySH)>thenoyltrifluoroacetone (TTAH)>acetylacetone (AcAcH). The optimal extraction pressure and temperature for the supercritical CO(2) extraction of gallium (III) with chelating agent PySH were found to be 70 degrees C and 3000psi, respectively. The optimum concentration of the chelating agent was found to be 50ppm. A value of 7.5 was selected as the optimum dimensionless CO(2) volume. The optimum pH of the solution for supercritical CO(2) extraction should fall in the range of 2.0-3.0. PMID- 18406057 TI - Time saving modified BCR sequential extraction procedure for the fraction of Cd, Cr, Cu, Ni, Pb and Zn in sediment samples of polluted lake. AB - The mobility, availability and persistence of heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) in contaminated lake sediment samples were evaluated by means of sequential extraction scheme, proposed by the Community Bureau of Reference protocol (BCR). The metal content in the extracts was measured by atomic absorption spectrometry. The precision and accuracy of the proposed procedure were evaluated by using a certified reference material BCR 701. The maximum recoveries for heavy metals (HMs) were observed for all three steps of BCR protocol at 32h total shaking period instead of previously reported 51h, with p>0.05. The lixiviation tests (DIN 38414-S4) were used to evaluate the leaching of HMs from sediment samples and it was observed that levels of water extractable HMs were low as compared to those values obtained by acid-soluble fraction of the BCR protocol. PMID- 18406058 TI - Column study of chromium(VI) adsorption from electroplating industry by coconut coir pith. AB - The removal of Cr(VI) from electroplating wastewater by coir pith was investigated in a fixed-bed column. The experiments were conducted to study the effect of important parameters such as bed depth (40-60cm) and flow rate (10-30ml min(-1)). At 0.05 C(t)/C(0), the breakthrough volume increased as flow rate decreased or a bed depth increased due to an increase in empty bed contact time (EBCT). The bed depth service time model (BDST) fit well with the experimental data in the initial region of the breakthrough curve, while the simulation of the whole curve using non-linear regression analysis was effective using the Thomas model. The adsorption capacity estimated from the BDST model was reduced with increasing flow rate, which was 16.40mg cm(-3) or 137.91mg Cr(VI)g(-1) coir pith for the flow rates of 10ml min(-1) and 14.05mg cm(-3) or 118.20mg Cr(VI)g(-1) coir pith for the flow rates of 30ml min(-1). At the highest bed depth (60cm) and the lowest flow rate (10mlmin(-1)), the maximum adsorption reached 201.47mg Cr(VI)g(-1) adsorbent according to the Thomas model. The column was regenerated by eluting chromium using 2M HNO(3) after adsorption studies. The desorption of Cr(III) in each of three cycles was about 67-70%. The desorption of Cr(III) in each cycle did not reach 100% due to the fact that Cr(V) was present through the reduction of Cr(VI), and was still in coir pith, possibly bound to glucose in the cellulose part of coir pith. Therefore, the Cr(V) complex cannot be desorbed in solution. The evidence of Cr(V) signal was observed in coir pith, alpha-cellulose and holocellulose extracted from coir pith using electron spin resonance (ESR). PMID- 18406059 TI - Melatonin inhibits amphetamine-induced increase in alpha-synuclein and decrease in phosphorylated tyrosine hydroxylase in SK-N-SH cells. AB - alpha-Synuclein is an abundant presynaptic protein implicated in neuronal plasticity and neurodegeneration disorders. Understanding alpha-synuclein function in dopaminergic cells could add to our knowledge of this key protein which is implicated in Parkinson's disease. Chronic or intermittent amphetamine (AMPH) abuse may create temporary or permanent disturbances in the dopaminergic system of the brain that may predispose individuals to Parkinsonism. Our previous studies showed that neurotoxicity induced by AMPH was mediated by enhanced oxidative stress and these effects were abolished by melatonin, a main secretory product of pineal gland. The present study was conducted to investigate the effect of AMPH on alpha-synuclein in regulating tyrosine hydroxylase (TH), a rate limiting enzyme for dopamine synthesis, in cultured human dopaminergic SK-N-SH cells. Of these, phosphorylation of Ser40 (pSer40) contributes significantly to TH activation and dopamine synthesis. Our data indicated that AMPH significantly increased the level of alpha-synuclein to 183% of the control value while reducing the levels of phosphorylated TH (TH-pSer40) enzyme and mitochondrial complex I to 78 and 52.9% of the control values, respectively and these effects were attenuated by melatonin. Further studies are needed to explore the mechanism by which alpha-synuclein contributes to TH-pSer40 dephosphorylation and the mechanism by which melatonin contributes to this interaction. PMID- 18406060 TI - Objects and their icons in the brain: the neural correlates of visual concept formation. AB - We are constantly exposed to symbols such as traffic signs, emoticons in internet communication, or other abstract representations of objects as well as, of course, the written words. However, aside from the word reading, little is known about the way our brain responds when we read non-lexical iconic symbols. By using functional MRI, we found that the watching of icons recruited manifold brain areas including frontal and parietal cortices in addition to the temporo occipital junction in the ventral pathway. Remarkably, the brain response for icons was contrasted with the response for corresponding concrete objects with the pattern of 'hyper-cortical and hypo-subcortical' brain activation. This neural underpinning might be called the neural correlates for visual concept formation. PMID- 18406061 TI - Amblyomma variegatum in cattle in Marie Galante, French Antilles: prevalence, control measures, and infection by Ehrlichia ruminantium. AB - We report Marie Galante as one of the Caribbean islands most heavily infested by the tropical bont tick (TBT) Amblyomma variegatum which is associated with two major diseases of ruminants: heartwater and dermatophilosis. In 2005, a survey was undertaken to assess the prevalence of TBT infestation in cattle, the prevalence of Ehrlichia ruminantium infection in TBTs, and the tick control measures implemented by livestock owners. A random sample of 195 cattle herds out of 1885 recorded on the island was investigated by thoroughly counting adult ticks on each animal and filling a questionnaire. A randomly collected sample of 136 TBTs was tested for infection by E. ruminantium by pCS20 nested PCR. Cattle herd prevalence (hp) was 73.8% for infestation by at least one TBT, 17.9% for infestation by at least one engorged female TBT, and 8.2% for clinical dermatophilosis. Cattle individual prevalence was 42.3% for infestation by at least one TBT, 6.6% for infestation by at least one engorged female TBT, and 2.2% for clinical dermatophilosis. The minimum, maximum and average numbers of TBTs per infested animal were, respectively 1, 108 and 11.5. Prevalence of TBT infection by E. ruminantium was 19.1%. No significant difference in herd prevalence was found among parishes or among ecological zones. For cattle owners treating against ticks (97.9% of all owners), all used aspersion of amitraz and herd prevalence was significantly different among those treating every 1-2-week (hp=69.6%, n=148), and less often than every 2-week (hp=88.6%, n=35) (P=0.031). Of the 42 herd subunits treated less than 4 days before the survey, 27 (64%) were infested with at least one TBT, and 6 (14%) with at least one engorged female TBT. These results indicate a high level of TBT infestation in Marie Galante, the inefficacy of tick treatments currently performed, and the need for an improved tick control strategy. Persisting high levels of infestation in Marie Galante threaten the success of on-going TBT eradication programs in the Caribbean because TBT can spread through migrating birds and trade of animals or of animal hides to other islands and potentially the American continent. PMID- 18406062 TI - Differential activation of mitogen-activated protein kinase signalling pathways in the hippocampus of CRND8 transgenic mouse, a model of Alzheimer's disease. AB - Transgenic Centre for Research in Neurodegenerative Diseases 8 (TgCRND8) mice expressing a double mutant form of human amyloid precursor protein represent a good model of Alzheimer's disease, and can be useful to clarify the involvement of mitogen-activated protein kinases (MAPK) dysregulation in the pathophysiology of this neurodegenerative disorder. Activation of extracellular regulated kinase (ERK) 1/2, jun kinase (JNK) and p38MAPK was studied in the hippocampus of 7-month old TgCRND8 mice by immunohistochemistry and Western blot analysis using antibodies selective for the phosphorylated, and thus active, forms of the enzymes. We demonstrated that the three main MAPK pathways were differentially activated in cells of the hippocampus of TgCRND8 mice in comparison to wild type (Wt) littermates, p38MAPK and JNK being more activated, while ERK less activated. p38MAPK was significantly activated in microglia, astrocytes and neurons, around and distant from the plaques. JNK was highly activated in cells closely surrounding the plaques. No difference was observed in the activation of the two major bands of JNK, at a molecular weight of 46 kDa and 54 kDa. These data indicate the possible involvement of p38MAPK and JNK pathways dysregulation in the pathogenesis of Alzheimer's disease. The ERK2 isoform of the ERK pathway was less activated in the hippocampal dentate gyrus of Tg mice in basal conditions. Furthermore activation of the ERK pathway by ex vivo cholinergic stimulation with carbachol caused significantly higher activation of ERK in the hippocampus of Wt mice than in Tg mice. These findings may pose a molecular basis for the memory disruption of Alzheimer's disease, since proper functioning of the basal forebrain cholinergic neurons and of ERK2 is critical for memory formation. PMID- 18406063 TI - Endosomal trafficking of AMPA-type glutamate receptors. AB - Many different forms of synaptic plasticity have been shown to ultimately modulate the number of AMPA-type glutamate receptors at the synapse. This trafficking involves lateral movements between synaptic and extrasynaptic sites at the neuron surface, as well as vesicular transport between the plasma membrane and intracellular compartments. Several new studies have shed light on the location and regulation of AMPA-type receptor (AMPAR) endocytosis, their intracellular sorting to divergent pathways at the level of endosomes, and the mechanism and sites of receptor recycling. This review summarizes this recent data on the trafficking along the endocytic pathway, and follows the path of internalized AMPAR from endocytosis up to sites of recycling. PMID- 18406064 TI - Task-dependent encoding of space and events by striatal neurons is dependent on neural subtype. AB - The dorsal striatum plays a critical role in procedural learning and memory. Current models of basal ganglia assume that striatal neurons and circuitry are critical for the execution of overlearned, habitual sequences of action. However, less is known about how the striatum encodes task information that guides the performance of actions in procedural tasks. To explore the striatal encoding of task information, we compared the behavioral correlates of striatal neurons tested in two tasks: a multiple T-maze task in which reward delivery was entirely predictable based on spatial cues (the Multiple-T task), and a task in which rats ran on a rectangular track, but food delivery depended on the distance traveled on the track and was not dependent solely on spatial location (the Take-5 task). Striatal cells recorded on these tasks were divisible into three cell types: phasic-firing neurons (PFNs), tonically firing neurons (TFNs), and high-firing neurons (HFNs) and similar proportions of each cell type were found in each task. However, the behavioral correlates of each cell type were differentially sensitive to the type of task rats were performing. PFNs were responsive to specific task-parameters on each task. TFNs showed reliable burst-and-pause responses following food delivery and other events that were consistent with tonically active neurons (TANs) on the Take-5 (non-spatial) task but not on the Multiple-T (spatial) task. HFNs showed spatial oscillations on the Multiple-T (spatial) task but not the Take-5 (non-spatial) task. Reconstruction of the rats' position on the maze was highly accurate when using striatal ensembles recorded on the Multiple-T (spatial) task, but not when using ensembles recorded on the Take-5 (non-spatial) task. In contrast, reconstruction of time following food delivery was successful in both tasks. The results indicated a strong task dependency of the quality of the spatial, but not the reward-related, striatal representations on these tasks. These results suggest that striatal spatial representations depend on the degree to which spatial task-parameters can be unambiguously associated with goals. PMID- 18406066 TI - Psychological stress, insulin resistance, inflammation and the assessment of heart disease risk. Time for a paradigm shift? AB - There is growing evidence that the present risk assessment protocol for coronary heart disease appears to underestimate the risk in general and the presence and progression of atherosclerosis in particular. Little or no correlation has been found between the 10-year risk based on the Framingham model and the extent or progression of coronary calcification. In addition, a number of studies find the protocol based on current guidelines leads to an under appreciation of the risk of symptomatic coronary heart disease or the associated fatal and non-fatal events, especially in younger asymptomatic individuals and women. Furthermore, the current guidelines give secondary importance to insulin resistance and inflammation and do not include psychosocial stress and depression, both of which are established and important risk factors for coronary heart disease. An alternative approach to risk assessment is proposed which emphasizes insulin resistance and psychological stress and depression and gives much greater recognition to inflammation as a root cause and target for intervention than is found in current guidelines. Consistent with this view, a revised assessment protocol is suggested which is still appropriate to the primary care setting and which might provide a different and perhaps more effective and relevant approach to primary prevention and risk reduction. PMID- 18406065 TI - Effect of microdialysis perfusion of 4,5,6,7-tetrahydroisoxazolo-[5,4-c]pyridine 3-ol in the perifornical hypothalamus on sleep-wakefulness: role of delta-subunit containing extrasynaptic GABAA receptors. AB - Gaboxadol or 4,5,6,7-tetrahydroisoxazolo-[5,4-c]pyridine-3-ol (THIP) is a selective agonist for the delta-subunit containing extrasynaptic GABA(A) receptors that will soon enter the U.S. market as a sleep aid [Winsky-Sommerer R, Vyazovskiy VV, Homanics GE, Tobler I (2007) The EEG effects of THIP (gaboxadol) on sleep and waking are mediated by the GABA(A)delta-subunit-containing receptors. Eur J Neurosci 25:1893-1899]. Numerous studies have shown that systemic administration of THIP reduces wakefulness and increases sleep both in humans and rats [Lancel M, Langebartels A (2000) Gamma-aminobutyric acid(A) (GABA(A)) agonist 4,5,6,7-tetrahydroisoxazolo[4,5-c]pyridin-3-ol persistently increases sleep maintenance and intensity during chronic administration to rats. J Pharmacol Exp Ther 293:1084-1090; Walsh JK, Deacon S, Dijk DJ, Lundahl J (2007) The selective extrasynaptic GABAA agonist, gaboxadol, improves traditional hypnotic efficacy measures and enhances slow wave activity in a model of transient insomnia. Sleep 30:593-602]. However, it is yet unclear where in the brain THIP acts to promote sleep. Since the perifornical lateral hypothalamus (PFH) contains orexin neurons and orexin neurons are critical for maintenance of arousal [McCarley RW (2007) Neurobiology of rapid eye movement (REM) and NREM sleep. Sleep Med 8:302-330], we hypothesized that THIP may act on PFH neurons to promote sleep. To test our hypothesis, we used reverse microdialysis to perfuse THIP unilaterally into the PFH and studied its effects on sleep-wakefulness during the light period in freely behaving rats. Microdialysis perfusion of THIP (100 microM) into the PFH produced a significant reduction in wakefulness with a concomitant increase in non-rapid eye movement or slow wave sleep as compared with artificial cerebrospinal fluid perfusion. REM sleep was unaffected. This is the first study implicating the delta-subunit containing extrasynaptic GABA(A) receptors in PFH in control of sleep-wakefulness in freely behaving rats. PMID- 18406067 TI - Delay of diagnosis and treatment of colorectal cancer--a population-based Danish study. AB - BACKGROUND: Recently, the authors have shown a doubled risk of having an advanced rectal cancer (RC) (Dukes' stage C or D) at the time of treatment, if the interval between onset of symptoms and start of treatment (treatment delay) was >60 days [Korsgaard M, Pedersen L, Sorensen HT, Laurberg S. Treatment delay is associated with advanced stage of rectal cancer but not of colon cancer. Cancer Detect Prev 2006;30(4):341-6]. The authors examined the treatment delay for colorectal cancer (CRC), as influenced by the patients, the general practitioners (G.P.), and the hospitals. METHOD: Population-based prospective observational study based on 743 Danish CRC-patients. Treatment delay was determined through questionnaire interviews. We examined the patient delay, the G.P. delay, and the hospital delay, and thereby the frequency of patients for whom the Danish fast track recommendations of a maximum of 14 days to diagnose CRC, and 14 days from the diagnosis to start the of treatment, were met. Colon cancer (CC) and RC patients were analyzed separately. RESULTS: Patient delay, in particular, was long, and longest for RC-patients (median 44 days vs.18 days). Median G.P. delay was short, but 25% of the CC-patients had a G.P. delay of 59 days or more, and 25% of the RC-patients had a G.P. delay of 53 days or more. The fast-track recommendations were poorly met; 53% of the CC-patients and 39% of the RC patients waited >14 days after referral for the diagnosis. 29% of the CC patients, and 53% of the RC-patients waited >14 days before the start of treatment. CONCLUSION: The total delay was too long, and can be shortened by optimizing all delay intervals. PMID- 18406068 TI - Descriptive epidemiology of esophageal carcinoma in the Ohio Cancer Registry. AB - BACKGROUND: Etiologic factors and demographics in esophageal cancer have not been fully characterized at a population-level. This study aimed to compare incidence rates of esophageal adenocarcinoma (EAC) and squamous cell carcinoma (ESCC) by race. Other aims were to evaluate the impact of race, age, gender, and histology on presenting stage, and to describe tobacco use history in EAC as documented in a cancer registry. METHODS: Invasive esophageal cancer cases reported to Ohio's Cancer Registry 1998-2002 were identified. Incident staged EAC and ESCC cases were analyzed for factors associated with metastatic disease. RESULTS: 930 ESCC and 1801 EAC cases were identified. African-Americans had higher ESCC incidence than whites (5.0 versus 1.3 cases/100,000/year). However, whites had higher EAC incidence (3.3 versus 0.8 cases/100,000/year). 77% of EAC cases with available tobacco history were reported in tobacco users. In univariate analyses, race, age, gender, and histology differed significantly by stage. 31% of patients aged > or =65 presented with distant stage, versus 26% of those <65 (p<0.001). 32% of African-Americans had distant stage, versus 34% of whites (p=0.048). In logistic regression modeling, male gender [OR 1.76, CI (1.15, 2.67)] and age <75 [OR 1.95, CI (1.21, 3.15)], but not race, predicted distant stage ESCC. Distant stage EAC was associated with age <56 [OR 1.82, CI (1.39, 2.38)] but not significantly associated with African-American race (p=0.062) for the sample size available. CONCLUSIONS: Whites had higher EAC rates, and African-Americans had higher ESCC rates. African-Americans were not more likely than whites to present with metastatic ESCC. PMID- 18406069 TI - Human papillomavirus, cervical cancer and women's knowledge. AB - BACKGROUND: Human papillomavirus (HPV) is the major risk factor for cervical cancer. METHODS: We implemented a retrospective case-series study to discern HPV knowledge accuracy among women diagnosed with and treated for cervical cancer. Cases (n=1174), identified from the Pathology database, were diagnosed and treated for cervical cancer at the same institution. Data were collected using self-administered questionnaires and by reviewing medical records. RESULTS: A total of 328 (27.9%) women returned the completed forms. Only 19% of the respondents had identified HPV as the primary risk factor for cervical cancer. Environmental pollutants, radiation exposure, poor dietary habits, excessive physical activity and family history of cervical cancer were listed as risk factors among many others. Multivariate analysis was performed to determine variables that were best associated with HPV knowledge accuracy. Age and education were the two variables that were statistically associated with the outcome. Younger and more educated women who participated in this study were more likely to know about the association between HPV infection and the risk of cervical cancer. CONCLUSIONS: Cervical cancer risk factor knowledge, especially knowledge about HPV is low, even among women with the history of cervical cancer. Younger and more educated women are more likely to have HPV and cervical cancer knowledge accuracy. The importance of personal health practices and the focus on health education should be equally emphasized to achieve successful cancer prevention through vaccination. PMID- 18406070 TI - MGN-3/Biobran, modified arabinoxylan from rice bran, sensitizes human breast cancer cells to chemotherapeutic agent, daunorubicin. AB - BACKGROUND: MGN-3/Biobran, a modified form of arabinoxylan from rice bran, is a potent biological response modifier (BRM). Our previous studies demonstrated that MGN-3 sensitizes human leukemia cells to death receptor [CD95]-induced apoptosis [Ghoneum M, Gollapudi S. MGN-3 sensitizes human T cell leukemia cells to death receptor (CD95)-induced apoptosis. Cancer Lett 2003;201:41-9]. In this study, we evaluated the chemo-sensitizing activity of MGN-3 against human breast cancer cells (BCCs) in vitro. METHODS: BCCs (MCF-7 and HCC70 cells) were cultured with different concentrations of daunorubicin (DNR) (from 1x10(-9) to 1x10(-6)M) in the presence or absence of selected concentrations of MGN-3 (100-1000mug/ml) for 3 days. Cancer cell survival was determined by MTT assay and drug accumulation was determined by flow cytometry. RESULTS: Treatment with MGN-3 increased susceptibility of BCCs to DNR (5.5-fold for MCF-7 and 2.5-fold for HCC70 cells) as compared to BCCs treated with DNR alone. The sensitizing effect of MGN-3 was associated with increased accumulation of DNR in cancer cells. CONCLUSIONS: Our data demonstrate that MGN-3 is an effective chemo-sensitizer and may represent a potential novel adjuvant for the treatment of breast cancer. PMID- 18406071 TI - [Lidocaine intoxication in newborn following maternal pudendal anesthesia: report of three cases]. AB - Pudendal nerve block may be indicated during instrumental delivery in situations where peridural anesthesia is unavailable. We report three cases of neonatal lidocaine intoxication following maternal pudendal block during delivery. Clinical features were hypotonia, pupillary mydriasis fixed to light, apnea, cyanosis and seizures. Two neonates required mechanical ventilation. Lidocaine was found in the serum of two babies. In all three cases, recovery was complete. The pharmacokinetics of lidocaine in a highly vascularized perineum during labor increase the risk of neonatal intoxication. A possible intoxication by local anesthetics should be considered in neonates presenting an acute distress in the delivery room. PMID- 18406072 TI - [Minutes of the EBCOG General Assembly in Vienna, December 2007]. PMID- 18406073 TI - [Minutes of the EBCOG General Assembly in Lisbon, March 2008]. PMID- 18406074 TI - [Duration of pushing in labor: literature review]. AB - French guidelines recommend performing an assisted delivery after 30min pushing, even if the fetal heart rate is reassuring. A literature review shows that international practices differ from French recommendations; primiparous women usually push on average one hour. Specific data about duration of the active second stage are scarce. Therefore, there is no evidence that maternal and neonatal outcome would be improved by an early-assisted delivery as compared to pushing efforts prolonged after 30min. PMID- 18406075 TI - The exploratory behaviour of rats in the hole-board apparatus: is head-dipping a valid measure of neophilia? AB - The exploratory behaviour of laboratory rodents is of interest within a number of areas of behavioural pharmacology. However, how best to measure exploratory behaviour in rodents remains a contentious issue. Many unconditioned tests, such as the open field, potentially confound general locomotor activity with exploration. The hole-board apparatus appears to avoid this confound, as head dipping into holes in the floor is assumed to be a valid measure of the subject's attraction towards novelty (neophilia). This study aimed to investigate whether head-dipping should be considered a valid measure of neophilia by comparing performance of adult male and female Lister hooded rats on the hole-board task (a) over repeated sessions and (b) when novel objects were absent or present underneath the holes. The results show that head-dipping initially decreased across repeated exposures, while time spent in the aversive central area increased. No change in head-dipping was seen in response to objects being placed underneath the holes. Rather than being a measure of neophilia, these results support the hypothesis that head-dipping represents an escape response, which declines as the subject becomes less fearful. These results are compared with previous studies of repeated exposure to other novel environments. PMID- 18406076 TI - The parasitic nematode Phasmarhabditis hermaphrodita defends its slug host from being predated or scavenged by manipulating host spatial behaviour. AB - Infective stages of commercially used molluscicidal rhabditide nematodes Phasmarhabditis hermaphrodita contain bacterial symbionts which kill their host by septicaemia. The nematodes feed on the multiplying bacteria and entire host tissue, develop and repeatedly reproduce. Invertebrate cadavers are rapidly (from minutes to hours) removed by scavengers. However nematodes need days to complete their life cycle inside the host. The post mortem locations of slugs killed by six different treatments (three types of molluscicides, a simulation of unsuccessful predation and two P. hermaphrodita nematode treatments) were compared. In comparison to other pathogenic states, significantly more slugs killed by the nematodes died within the soil, where the scavenging pressure is weaker than on the soil surface (where most of the slugs died regardless treatment). We suggest that this is an outcome of behavioural manipulation, which prevent the parasites from being predated or scavenged together with their host until the nematodes complete development inside the host cadaver. PMID- 18406077 TI - Dynamics of choice and frequency of environment change. AB - Recent research has examined the effects on behavioral adjustment of a range of within-session changes in the reinforcer ratio available from two alternatives. Regularities between extended patterns of behavior (choice) and a distribution of food deliveries (expressed as a food-rate ratio) have emerged from such studies. To assess the generality of these findings, the present study manipulated rats' experience in the choice situation. Sixteen rats were divided into two groups of eight rats each. Seven non-signaled food-rate ratios were arranged to occur for responses on two concurrently available levers. Group 1 experienced a different food-rate ratio every day; each ratio provided 70 food deliveries, and none of the seven ratios operated for 2 consecutive days. Group 2 experienced the same sequence of food-rate ratios, except that each food-rate ratio remained in effect for 17 consecutive days (providing 70 food deliveries per session), after which a different food-rate ratio was selected for the next 17 sessions. In all cases, preference was adjusted to changes in food-rate ratios. Estimates of sensitivity to changes in food-rate ratios increased with an increasing number of sessions (i.e., experience in the choice situation). PMID- 18406078 TI - Effect of contingent auditory stimuli on concurrent schedule performance: an alternative punisher to electric shock. AB - This study explored whether load auditory stimuli could be used as functional punishing stimuli in place of electric shock. Three experiments examined the effect of a loud auditory stimulus on rats' responding maintained by a concurrent reinforcement schedule. In Experiment 1, overall response rate decreased when a concurrent 1.5 s tone presentation schedule was superimposed on the concurrent variable interval (VI) 180-s, VI 180-s reinforcement schedule. On the contrary, response rate increased when a click presentation schedule was added. In Experiment 2, the extent of the response suppression with a 1.5 s tone presentation varied as a function of the frequency of the reinforcement schedule maintaining responses; the leaner the schedule employed, the greater the response suppression. In Experiment 3, response suppression was observed to be inversely related to the duration of the tone; response facilitation was observed when a 3.0-s tone was used. In Experiments 1 and 2, a preference shift towards the alternative with richer reinforcement was observed when the tone schedule was added. In contrast, the preference shifted towards the leaner alternative when the click or longer duration stimulus was used. These results imply that both the type and duration of a loud auditory stimulus, as well as the reinforcement schedule maintaining responses, have a critical role in determining the effect of the stimuli on responding. They also suggest that a loud auditory stimulus can be used as a positive punisher in a choice situation for rats, when the duration of the tone is brief, and the reinforcement schedule maintaining responses is lean. PMID- 18406079 TI - Comparative psychometric study of a range of hazardous drinking measures administered online in a youth population. AB - AIMS: To compare the psychometric performance of a range of existing alcohol measures when data are collected online with young people, and thereby to gain insights into the reliability and validity of this mode of data collection. METHOD: One hundred and sixty-seven U.K. resident young people aged 16-24 who had drunk alcohol within the past week participated in a cross-sectional psychometric study with a test-retest reliability component. Eight hazardous drinking measures were used: the alcohol use disorders identification test (AUDIT) summary instrument and dedicated assessments of consumption (timeline follow-back and diary-format recall of alcohol drunk in the last 7 days), dependence (Leeds dependence questionnaire and severity of dependence scale) and problems (Rutgers alcohol problem index, alcohol problems scale and academic role expectations and alcohol scale). RESULTS: Internal consistency and test-retest correlation statistics were generally satisfactory, providing evidence of reliability. Validation data obtained in principal components analyses, investigation of the correlation matrix and in a multiple regression model of total AUDIT score were also supportive of the online use of these measures. Evidence was weakest for the alcohol problems scale. CONCLUSIONS: A range of hazardous drinking measures exhibit sound psychometric properties when administered online. Further comparative study of the relationships between different measures is needed. PMID- 18406080 TI - Binding of epsilon-toxin from Clostridium perfringens in the nervous system. AB - Epsilon-toxin (epsilon-toxin), produced by Clostridium perfringens type D, is the main agent responsible for enterotoxaemia in livestock. Neurological disorders are a characteristic of the onset of toxin poisoning. Epsilon-Toxin accumulates specifically in the central nervous system, where it produces a glutamatergic mediated excitotoxic effect. However, no detailed study of putative binding structures in the nervous tissue has been carried out to date. Here we attempt to identify specific acceptor moieties and cell targets for epsilon-toxin, not only in the mouse nervous system but also in the brains of sheep and cattle. An epsilon-toxin-GFP fusion protein was produced and used to incubate brain sections, which were then analyzed by confocal microscopy. The results clearly show specific binding of epsilon-toxin to myelin structures. epsilon-Prototoxin GFP and epsilon-toxin-GFP, the inactive and active forms of the toxin, respectively, showed identical results. By means of pronase E treatment, we found that the binding was mainly associated to a protein component of the myelin. Myelinated peripheral nerve fibres were also stained by epsilon-toxin. Moreover, the binding to myelin was not only restricted to rodents, but was also found in humans, sheep and cattle. Curiously, in the brains of both sheep and cattle, the toxin strongly stained the vascular endothelium, a result that may explain the differences in potency and effect between species. Although the binding of epsilon-toxin to myelin does not directly explain its neurotoxic effect, this feature opens up a new line of enquiry into its mechanism of toxicity and establishes the usefulness of this toxin for the study of the mammalian nervous system. PMID- 18406081 TI - Isolation of a novel viral agent associated with porcine reproductive and neurological syndrome and reproduction of the disease. AB - Disease outbreaks characterized by reproductive failure and/or neurologic disorders, which are commonly referred as "Porcine Reproductive and Neurologic Syndrome (PRNS)", were observed in many swine farms in Iowa and other states. Although an infectious cause was suspected to account for the disease, no conclusive diagnosis had been reached with respect to conventional infectious agents. Extensive laboratory diagnostic investigation on suspect cases repeatedly resulted in the isolation of a cytopathic enveloped virus of 50-60nm in size from nervous and second lymphoid tissues and sera and, to reflect its unknown identity, named "Virus X". The presence of virus particle with morphological characteristics similar to Virus X in tissues from affected animals was also observed on thin-section positive-staining electron microscopy. Isolates of Virus X were not readily recognized by antibodies raised against any known viruses pathogenic to swine but by antisera collected from animals surviving clinical episode, indicating that Virus X is likely a previously unrecognized agent. Pregnant sows experimentally inoculated with Virus X (ISUYP604671) or homogenate (filtrate) of tissues from a clinically affected animal developed clinical signs and pathological changes similar to field observations including the loss of pregnancy. Furthermore, caesarian-derived, colostrum-deprived young pigs developed mild encephalomyelitis lesions in brains after experimental inoculation with the virus or the tissue homogenate although clinical neurologic signs were not observed. More importantly, Virus X was re-isolated from all inoculated animals while control pigs remained negative for the virus during the study. Collectively, Virus X is a novel viral agent responsible for PRNS and remains to be further characterized for taxonomical identity. PMID- 18406082 TI - Steroid secretion following exposure of ovarian follicular cells to single congeners and defined mixture of polybrominateddibenzoethers (PBDEs), p,p'-DDT and its metabolite p,p'-DDE. AB - The main objective of the present study was to describe in vitro effects of individual congeners of polybrominateddibenzoethers (PBDEs) (PBDE-47, 99, 100, 209), p,p'-dichlorodiphenyltrichloroethane (DDT) and p,p' dichlorodiphenyldichloroethylene (DDE) present at the highest amount in real-life mixtures of POPs extracted from liver oil of burbot from Lake Mjosa in Norway, and defined combinations of these compounds on steroidogenesis in porcine ovarian follicular cells. Co-culture of theca and granulosa cells (Gc) was exposed by individual congeners of PBDEs or mixture. Media samples were collected and used for cell viability and steroid determination, while cells were used for the measurement of caspase-3 activity. All investigated PBDEs congeners increased both testosterone (T) and estradiol (E2) secretion. We noted additive stimulatory action of BDE-47 and BDE-100 on both T and E2 secretion, and then inhibitory action on both of these hormones production when additionally BDE-99 and then BDE 209 were included in this mixture. The situation was different when DDT and DDE were added to the defined mixture. Stimulatory action on E2 secretion with parallel decrease of T secretion was noted. In conclusion, the data indicated: (1) stimulatory action of individual congeners of PBDEs on both E2 and T secretion by ovarian follicular cells points to stimulatory action on enzymes responsible for testosterone secretion; (2) the overall effect of a real-life mixture on the steroidogenic response in ovarian follicular cells could be dependent on the concentration of DDTs in the mixture; (3) the predicted effect of mixture, calculated based on a sum of independent action of individual congeners, was higher than that noted under the influence of defined mixture. PMID- 18406083 TI - Body mass index trajectories and age at menopause in a British birth cohort. AB - OBJECTIVE: This study investigates the influence of body mass index (BMI) at ages 15, 20, 26, 36, and 43, and of BMI trajectories from 20 to 36 years on the timing of menopause and hormone therapy (HT) use until age 57 years. METHODS: A nationally representative British cohort of 1583 women born in March 1946 with prospective data across the life course. RESULTS: By age 57, a total of 695 women had experienced natural menopause while 431 women had started HT prior to menopause. Cox regression models indicated no significant associations between BMI at any age, or BMI trajectory, and timing of natural menopause. At every age BMI was strongly (p< or =0.01) and linearly associated with age at HT use and BMI from 26 years onwards was associated with age at first event (menopause or HT use). Decreasing BMI was associated with earlier HT use at all ages. These associations were not accounted for by parity, cigarette smoking or childhood and adult social class. CONCLUSION: BMI across the reproductive lifespan did not influence age at menopause to an extent that would be clinically relevant for postmenopausal health. Lower BMI at all ages and underweight trajectory were related to an earlier start of HT. Further studies are required to understand whether such relationships are due to underweight women experiencing menopause earlier (and because of menopausal symptoms starting HT earlier) than heavier women, or having behavioural characteristics related to earlier HT use, independent of menopause. PMID- 18406084 TI - Physicochemical effects of terpenes on organogel for transdermal drug delivery. AB - It is accepted that terpenes are effective penetration enhancers to promote the passage of drugs or chemicals through the human skin barrier. However the physical and chemical changes of a pharmaceutical vehicle induced by the incorporation of terpenes have not been explored. Thus, this study examines the effects of three terpenes (linalool, cineole, limonene) on the rheology and chemical stability of an organogel composed of dibutyllauroylglutamide (GP1) and propylene glycol (PG). At a given GP1 concentration, oxygen-containing linalool and cineole decreased gel moduli (elastic and viscous) and brittleness, and the reverse was obtained for hydrocarbon limonene. Probably, linalool and cineole interfered with hydrogen bonding between GP1 molecules while limonene could have initiated a phase separation-mediated gelation, changing the gel morphology. Microcalorimetry detected minute heat endotherms for gels (with and without terpenes) subjected to accelerated heat testing. These heat changes could arise from a small degree of structural disruption of the gel network. Heat endotherms normalized with respect to GP1 content were used to assess gel chemical stability. Although the terpenes altered rheology, they did not significantly affect the chemical stability of the gels. This is the first in the literature that reports the effect of penetration enhancers, such as terpenes, on the physical, rheological and chemical characteristics of a model pharmaceutical formulation for topical and transdermal drug delivery. PMID- 18406085 TI - Factors influencing hydrocortisone permeation into human hair follicles: use of the skin sandwich system. AB - The aim of the present study was to use the in vitro human skin sandwich system in order to quantify the influence of formulation variables on intrafollicular hydrocortisone permeation. The investigated variables were the pH and the viscosity of the topical formulation as well as the presence of chemical enhancers (carvone, menthone, oleic acid and sodium lauryl sulphate). Furthermore, skin sandwich hydration was also varied in order to determine if the method itself can be run using only partially hydrated skin tissues. It was determined that the follicular contribution to hydrocortisone flux decreased marginally with increasing alkalinity in the pH range 3-8.8. Intrafollicular penetration was markedly reduced when HPMC gels were used instead of an aqueous solution. Pretreating the skin with chemical enhancers also reduced the follicular contribution to flux, probably due to permeabilisation of the continuous stratum corneum. Furthermore, it was not possible to satisfactorily modify the skin sandwich method so that it could be deployed using less hydrated skin. PMID- 18406086 TI - Paclitaxel tumor biodistribution and efficacy after intratumoral injection of a biodegradable extended release implant. AB - PURPOSE: The aim of this study was to investigate the effectiveness of paclitaxel controlled release from intratumorally injected polymer. METHODS: The effectiveness of paclitaxel-polymer formulation injected intratumorally was tested in mouse bladder tumor model. To determine paclitaxel biodistribution in tumor at predetermined time periods the tumor was excised, frozen and sectioned, and the paclitaxel concentrations were determined in the tumor tissue and in plasma by HPLC. Histopathological evaluation of the necrosis and inflammation was performed on tumor sections. RESULTS: In the paclitaxel/polymer group mice were injected intratumorally with 0.2 ml of the 10% (w/w) paclitaxel formulation, the tumor disappeared completely 5 days after injection, and mice survived till the end of the study (50 days post-tumor cells inoculation). In biodistribution studies, the highest paclitaxel concentration in the tumor tissue was 40 microg/mg 1 day after the intratumoral injection and decreased gradually during 10 days to 5 microg/mg that is still high enough to induce cytotoxic effect, and the necrotic effect of paclitaxel on the tumors was confirmed by histopathology. CONCLUSIONS: Treatment with local injection of polymer-paclitaxel formulation inhibited the growth of solid tumors. Distribution studies of paclitaxel after intratumoral injection showed high and effective drug concentrations in tumor. PMID- 18406087 TI - Simultaneous synthesis and coating of salbutamol sulphate nanoparticles with L leucine in the gas phase. AB - Salbutamol sulphate nanoparticles have been simultaneously prepared and coated with L-leucine in the gas phase. Three different ways of coating can be separated based on the operation temperatures used in an aerosol flow reactor. Below the temperature of L-leucine sublimation, formation of the L-leucine layer on the core particle surface takes place via diffusion of L-leucine molecules on the droplet surfaces during droplet drying. At intermediate temperatures, the extent of sublimation of L-leucine depends notably on the concentration, and thus partial evaporation was expected. The L-leucine coating was solely formed via vapor deposition at high reactor temperatures when complete sublimation of L leucine was obtained. The geometric mean diameter of the core salbutamol particles was approximately 65 nm. In general, particle size increased with the addition of L-leucine. The size distribution remained the same or broadened when the coating layer of the particles was formed via surface diffusion whereas notable narrowing of the distribution was observed when the coating was formed via vapor deposition. Upon desublimation and heterogeneous nucleation on the surfaces of smooth, spherical core particles, L-leucine formed a discontinuous coating with leafy crystals a few nanometers in size. PMID- 18406088 TI - Pharmacokinetics of salvianolic acids after intravenous injection, with and without Panax quinquefolium protopanaxadiol saponins, in rats. AB - AIM: The purpose of the present study was to examine the effects of Panax quinquefolium protopanaxadiol saponins (PQDS) extracts on the plasma protein binding and pharmacokinetic of salvianolic acids extracts extracted from the traditional Chinese medical Salvia miltiorrhiza,. Salvianolic acids are used to treat myocardial ischemia, and PQDS has similar functions. It is expected to achieve a better therapeutic efficacy if the two extracts are developed as a compound prescription for injection. MATERIALS AND METHODS: An established high performance liquid chromatographic technique coupled with microdialysis was used. Male Sprague-Dawley rats were given salvianolic acids extracts and a compound of the two extracts via femoral vein. RESULTS: It was found that there were significant differences in the percentage protein binding as well as the pharmacokinetic parameters. The rat plasma protein binding of the four salvianolic acids increased by different degrees at three dose levels (25, 50, 100mg/kg of salvianolic acid B) when the two extracts were administered together. Also, their elimination half-life was prolonged, and their plasma concentrations remained stable longer after administration of a dose of 50mg/kg (salvianolic acid B). CONCLUSIONS: The results indicated that the PQDS extracts could delay the excretion of salvianolic acids as well as maintain the blood concentration higher than salvianolic acids extracts administered alone. PMID- 18406090 TI - Fatal intoxication with labetalol (Trandate). AB - The dead body of a 44-year-old woman, previously known for depression and alcoholism, has been discovered at her place of residence by her husband. A forensic autopsy has been carried out. The results indicated unspecific histological lesions (alveolar oedema, liver steatosis and interstitial nephritis) but did not reveal any apparent cause of death. Several boxes of medicines have been found near the body, justifying a toxicological analysis. This has been performed on peripheral blood and urine samples using liquid chromatography with diode array and mass spectrometric detections, in conjunction with gas chromatography coupled with mass spectrometry. Ethanol has been found (1.24 g/L in blood, 2.63 g/L in urine and 1.33 g/kg in gastric content), as well as therapeutic concentrations of meprobamate (14.1mg/L) and low concentrations of nordazepam (0.12 mg/L) in blood. On the other hand, particularly high levels of labetalol, a widely used beta-blocker, have been found both in blood (1.7 mg/L) and urine (20.2mg/L), which led us to measure labetalol levels in available viscera samples (liver, heart, kidney, and lung) and gastric content. Measured concentrations were 14.2 microg/g, 7.8 microg/g, 5.4 microg/g, 5.2 microg/g and 31.1 microg/g, respectively. We describe here the first report of a fatal intoxication attributed to labetalol that is linked to its acute toxicity, with tissue distribution of this beta-blocker. PMID- 18406091 TI - Colorectal carcinoma with potentially resectable metastases: factors associated with the failure of curative schedule. AB - BACKGROUND: The management of patients with colorectal cancer (CRC) and synchronous liver metastases (SLM) depends on the primitive tumor, resectability of the metastatic disseminations and the patient's comorbid condition(s). Considering all patients with potentially resectable primary CRC and SLM, curative resection (R0) will be possible in some patients, although in others surgery will never be performed. The purpose of our study was to identify factors of failure of the curative schedule in these patients. METHODS: We reviewed the data of patients with CRC and SLM between January 2002 and March 2007. Two groups were defined: group R0 when complete metastatic and primary tumor resection was finally achieved after one and more surgical stages and group R2 when curative resection was not possible at the end of the schedule. Clinical, pathologic and outcome data were retrospectively analyzed as well as preoperative management of SLM (chemotherapy, radiofrequency, portal vein embolization). RESULTS: Forty-five patients were included. Curative resection (group R0) was performed in 31 patients (69%) with 48% undergoing major hepatic resection. Mortality of hepatic resection was 0% although it was 9% for primitive tumor. Portal vein embolization was performed preoperatively in eight patients and radiofrequency ablation in 13. Median follow-up was 21 months. Overall survival was 86% at one year and 39% at three years. Survival in group 1 was 97 and 57% at one and three years respectively. Disease-free survival was 87 and 40% at one and three years. Tumor recurrence was noted in 61% of resected patients. At multivariate analysis, number of hepatic metastases superior than three and complicated initial presentation of primitive tumor were found to be significant and predictors of failure of hepatic resection. CONCLUSION: Aggressive management with curative resection of SLM may enable long-term survival. More than three SLM and complicated initial presentation of primitive tumor are factors predictive of failure of the curative schedule. PMID- 18406092 TI - (1)H MR spectroscopy of skeletal muscle, liver and bone marrow. AB - Proton magnetic resonance spectroscopy ((1)H MRS) offers interesting metabolic information even from organs outside the brain. In the first part, applications in skeletal muscle for determination of intramyocellular lipids (IMCL), which are involved in the pathogenesis of insulin resistance, are described. Peculiarities of spectral pattern are discussed and studies for short-term regulation of IMCL, as dietary intervention, exercise and fasting are presented. The second part deals with quantification of small amounts of lipids in the liver (hepatic lipids, HL), which is also of increasing interest in the field of diabetes research. Recommendations for correct assessment of spectra in this "moving organ" are given and the importance of HL is described by examples of a cohort at increased risk for type 2 diabetes. Regulation of HL is described on the basis of a few studies. The third part concentrates on spectral characterization of bone marrow. Peripheral bone marrow of adults consists mainly of fat, while central marrow regions in the pelvis, spinal column and breast bone (and the peripheral bone marrow of children as well) contribute to blood formation and show a variable composition of adipocytes (fat cells), interstitial fluid and water containing precursor cells for erythrocytes, leucocytes and thrombocytes. Adapted (1)H spectroscopic techniques allow a semi-quantitative analysis of bone marrow composition. PMID- 18406093 TI - Comparative analysis of the diversity of aerobic spore-forming bacteria in raw milk from organic and conventional dairy farms. AB - Bacterial contamination of raw milk can originate from different sources: air, milking equipment, feed, soil, faeces and grass. It is hypothesized that differences in feeding and housing strategies of cows may influence the microbial quality of milk. This assumption was investigated through comparison of the aerobic spore-forming flora in milk from organic and conventional dairy farms. Laboratory pasteurized milk samples from five conventional and five organic dairy farms, sampled in late summer/autumn and in winter, were plated on a standard medium and two differential media, one screening for phospholipolytic and the other for proteolytic activity of bacteria. Almost 930 isolates were obtained of which 898 could be screened via fatty acid methyl ester analysis. Representative isolates were further analysed using 16S rRNA gene sequencing and (GTG)(5)-PCR. The majority of aerobic spore-formers in milk belonged to the genus Bacillus and showed at least 97% 16S rRNA gene sequence similarity with type strains of Bacillus licheniformis, Bacillus pumilus, Bacillus circulans, Bacillus subtilis and with type strains of species belonging to the Bacillus cereus group. About 7% of all isolates may belong to possibly new spore-forming taxa. Although the overall diversity of aerobic spore-forming bacteria in milk from organic vs. conventional dairy farms was highly similar, some differences between both were observed: (i) a relatively higher number of thermotolerant organisms in milk from conventional dairy farms compared to organic farms (41.2% vs. 25.9%), and (ii) a relatively higher number of B. cereus group organisms in milk from organic (81.3%) and Ureibacillus thermosphaericus in milk from conventional (85.7%) dairy farms. One of these differences, the higher occurrence of B. cereus group organisms in milk from organic dairy farms, may be linked to differences in housing strategy between the two types of dairy farming. However, no plausible clarification was found for the relatively higher number of thermotolerant organisms and the higher occurrence of U. thermosphaericus in milk from conventional dairy farms. Possibly this is due to differences in feeding strategy but no decisive indications were found to support this assumption. PMID- 18406094 TI - Description of a novel indole-oxidizing bacterium Pseudomonas indoloxydans sp. nov., isolated from a pesticide-contaminated site. AB - A Gram-negative, deep brown-pigmented Gammaproteobacteria, strain IPL-1(T), capable of oxidizing indole was isolated from a lindane-contaminated site and subjected to a polyphasic taxonomic study. Most of the physiological and biochemical properties, major fatty acids (C(18:1)omega7c, C(16:1)omega7c/iso C(15:0) 2OH and C(16:0)), estimated DNA G+C content (67.2mol%) and 16S rRNA gene sequence analysis showed that strain IPL-1(T) belonged to the genus Pseudomonas. Strain IPL-1(T) exhibited highest 16S rRNA gene sequence similarity with Pseudomonas pseudoalcaligenes (99.0%), followed by Pseudomonas alcaliphila (98.7%), Pseudomonas oleovorans (98.3%), Pseudomonas nitroreducens (98.0%), Pseudomonas mendocina (97.6%) and Pseudomonas stutzeri (97.4%). However, the DNA DNA relatedness values between strain IPL-1(T) and the closely related taxa were between 22% and 61%. On the basis of differential phenotypic characteristics and genotypic distinctiveness, strain IPL-1(T) should be classified within the genus Pseudomonas as a novel species, for which the name Pseudomonas indoloxydans is proposed. The type strain is IPL-1(T) (=MTCC 8062(T)=JCM 14246(T)). PMID- 18406095 TI - Homocysteine activates calcium-mediated cell signaling mechanisms targeting the cytoskeleton in rat hippocampus. AB - Homocysteine is considered to be neurotoxic and a risk factor for neurodegenerative diseases. Despite the increasing evidences of excitotoxic mechanisms of homocysteine (Hcy), little is known about the action of Hcy on the cytoskeleton. In this context, the aim of the present work was to investigate the signaling pathways involved in the mechanism of action of Hcy on cytoskeletal phosphorylation in cerebral cortex and hippocampus of rats during development. Results showed that 100 microM Hcy increased the intermediate filament (IF) phosphorylation only in 17-day-old rat hippocampal slices without affecting the cerebral cortex from 9- to 29-day-old animals. Stimulation of (45)Ca(2+) uptake supported the involvement of NMDA receptors and voltage-dependent channels in extracellular Ca(2+) flux, as well as Ca(2+) release from intracellular stores through inositol-3-phosphate and ryanodine receptors. Moreover, the mechanisms underlying the Hcy effect on hippocampus cytoskeleton involved the participation of phospholipase C, protein kinase C, mitogen-activated protein kinase, phosphoinositol-3 kinase and calcium/calmodulin-dependent protein kinase II. The Hcy-induced IF hyperphosphorylation was also related to G(i) protein and inhibition of cAMP levels. These findings demonstrate that Hcy at a concentration described to induce neurotoxicity activates the IF-associated phosphorylating system during development in hippocampal slices of rats through different cell signaling mechanisms. These results probably suggest that hippocampal rather than cortical cytoskeleton is susceptible to neurotoxical concentrations of Hcy during development and this could be involved in the neural damage characteristic of mild homocystinuric patients. PMID- 18406096 TI - The power of community-based health communication interventions to promote cancer prevention and control for at-risk populations. PMID- 18406097 TI - Cancer coverage in a mainstream and Korean American online newspaper: lessons for community intervention. AB - OBJECTIVE: Korean Americans have higher incidence of stomach, liver, and cervical cancer than most groups in CA, USA, and generally lower rates of cancer screening. To better target community interventions, we evaluated the cancer content in a leading Korean American newspaper, the Korea Daily, compared to a mainstream newspaper from the same metropolitan area, the Los Angeles (LA) Times. METHODS: Using the online versions of each newspaper for the year 2006, we counted articles mentioning cancer and evaluated the content in a random sample of 300 articles from each newspaper. Articles were categorized by relevance of cancer content and topic(s) covered. RESULTS: Although the LA Times had a higher proportion of articles mentioning cancer, the Korea Daily had more articles that primarily focused on cancer and addressed specific types of cancer. Articles in the Korea Daily were more likely to discuss prevention, while those in the LA Times more often focused on people, politics, or research. CONCLUSIONS: Smaller, ethnic newspapers may be more amenable to messages about prevention and appropriate places to target community-focused interventions. PRACTICE IMPLICATIONS: Health practitioners should consider submitting articles to ethnic newspapers and forming partnerships with journalists to cultivate this potential. PMID- 18406098 TI - Enquiring minds want to know: topics requested by users of a children's health information website. AB - OBJECTIVE: To investigate health information needs of users of a health information website for children and young people. METHODS: A prospective analysis was conducted of unsolicited information requests to the Children First for Health (www.childrenfirst.nhs.uk) website, an NHS-run health and hospital online information resource for children, young people and families. All specific and general enquiries were recorded prospectively and coded according to health topic, age of the enquirer and web pages accessed. RESULTS: There were 2865 (62%) general health information and 924 (21%) specific enquiries received over an 18 month period (6.3 enquiries per day). More general enquiries were received about adolescent (64%) than children's (36%) health issues. The most common topics were psychosocial issues, hospital and health services, and normal growth and development. Only 1% of requests for information related to smoking, alcohol and drugs. CONCLUSION: Children and adolescents are active users of the Internet for health information. The results of this study highlight the health topics that are of greatest interest to children and young people. PRACTICE IMPLICATIONS: The findings provide objective data on health information topics of interest to UK children and young people and can be used to guide development of health information aimed at UK youth. PMID- 18406099 TI - Keeping morality out and the GP in. Consultations in Danish general practice as a context for smoking cessation advice. AB - OBJECTIVE: To describe consultations in Danish general practice as a context for a mass strategy of smoking cessation advice. METHODS: The focus of the study was on consultations for health problems that were not related to smoking. Interviews with eleven patients and their six GPs were grounded in observation of their own consultations. RESULTS: Patients and GPs agreed that the GP should adopt an attitude of moral acceptance towards patients. Ideals of moral acceptance of patients in general practice consultations were challenged by the prevailing negative moral values associated with smoking. A general aim of mutuality in the conversation in consultations could not always be achieved in smoking cessation advice. Achieving mutuality was especially a problem when smoking cessation advice was repeated at short intervals. CONCLUSION: Two elements of Danish general practice consultations were challenged by smoking cessation advice to patients without smoking-related illness: the ideal of moral acceptance of patients in general practice consultations held by GPs and patients and the wish for mutuality in the conversation during consultations. PRACTICE IMPLICATIONS: A conversation about smoking based on motivational interviewing would fit in the context of Danish general practice. Relieving the conversation of blocks due to moral implications, however, is still a challenge. PMID- 18406100 TI - Axillary recurrence after a tumour-negative sentinel node biopsy in breast cancer patients: A systematic review and meta-analysis of the literature. AB - BACKGROUND: Sentinel node biopsy became the standard of care before consensus on the technique was reached and without randomized studies having shown a similar or decreased axillary recurrence rate. The purpose of this study was to evaluate studies reporting on patients with a negative sentinel node biopsy. METHODS: We performed a systematic review and meta-analysis of the literature for studies concerning clinically node-negative breast cancer patients with a tumour-negative sentinel node biopsy and no subsequent axillary node dissection. The axillary recurrence rate was determined, as well as the sensitivity of the sentinel node procedure and the differences in lymphatic mapping techniques. RESULTS: Forty eight studies concerning 14 959 sentinel node-negative breast cancer patients followed for a median of 34 months were selected. Sixty-seven patients developed an axillary recurrence, resulting in a recurrence rate of 0.3%. The sensitivity of the sentinel node biopsy was 100%. Uni- and multivariable variable analyses showed that the lowest recurrence rates were reported in studies performed in cancer centres, in studies that described the use of (99m)Tc-sulphur colloid, and also when investigators used the superficial injection technique or evaluated the harvested sentinel nodes with haematoxylin-eosin and immunohistochemistry staining (p<0.01). CONCLUSIONS: In this systematic literature review, the axillary recurrence rate in sentinel node-negative patients is 0.3%, which is well within the desired range. The median sensitivity of the procedure appears to be as high as 100%. The recurrence rate is influenced by the differences in the lymphatic mapping technique. PMID- 18406101 TI - Editor's quinquennial report 2008. PMID- 18406102 TI - Vascular endothelial growth factor targeted RNA interference as a modulator of angiogenesis. AB - Angiogenesis is important in the pathogenesis of cancer and ischemic heart disease. Vascular endothelial growth factor targeted RNA interference is a very promising tool for modulation of blood vessel growth as recent studies show. This article tries to summarize the basic ideas behind the experimental use of RNA interference to regulate angiogenesis. PMID- 18406103 TI - Successful prevalidation of the slug mucosal irritation test to assess the eye irritation potency of chemicals. AB - A previous 'in house' validation study showed that the SMI assay can be used as an alternative to the in vivo Draize eye irritation test. The aim of this multi centre study with four participating laboratories was to assess the transferability and inter-laboratory variability of the assay using 20 reference chemicals covering the whole irritancy range. The eye irritation potency of the chemicals was assessed by measuring the amount of mucus produced during a 60-min contact period with a 1% dilution, and a second 60-min treatment with a 3.5% dilution. After each contact period the protein release from the mucosal surface was measured. Linear discriminant equations were used to convert the results into the corresponding EU eye irritation categories (NI, R36 and R41). All the non irritants were predicted correctly by the four laboratories resulting in a 100% specificity. For the R36 compounds a correct classification rate of 89% (VITO) and 100% (SPL, JNJ and UGent) was obtained. The R41 compounds were classified correctly in 78% of the cases for VITO, 89% for SPL and JNJ and 100% for UGent. We can conclude that the SMI assay is a relevant, easily transferable and reproducible alternative to predict the eye irritation potency of chemicals. PMID- 18406104 TI - Rapid, sensitive, and validated method for detection of Salmonella in food by an enrichment broth culture - nested PCR combination assay. AB - A rapid nested PCR assay for detection of Salmonella from food was developed. The sensitivity of the assay developed was comparable to the traditional culture based methods with an advantage in reduction of assay time. The assay procedure with artificially contaminated samples was able to detect as low as 4CFU Salmonella/25g of food samples (sprout, carrot, cucumber and poultry meat). With two synthetic primers of 26 mer TS11 and 25 mer TS4, a 1.2kb fragment was amplified which served as a template for amplification of final 375bp product using TS11 and TS5 primers. No non-specific amplification from the native microbial flora of food samples was observed. The reaction generates a single band specific to Salmonella which allows the analyst to interpret data at ease and without any confusion. Enriched broth serves as template for the reaction which removes labour intensive DNA isolation procedures. In case of artificially contaminated samples, 6h enriched lactose broth can serve as template. However, for market samples where the organisms are under environmental stress, it is desirable to use template from Rappaport Vasiliadis medium. The assay also employes internal amplification control, which is amplified into a 300bp fragment and thus serves as positive control for the reaction and any possibility of false negative due to inhibitory action of food components on PCR reaction can be ruled out. PMID- 18406105 TI - A model of contour extraction including multiple scales, flexible inhibition and attention. AB - A mathematical model of contextual integration and contour extraction in the primary visual cortex developed in a recent work [Ursino, M., & La Cara, G. E. (2004). A model of contextual interactions and contour detection in primary visual cortex. Neural Networks, 17, 719-735] has been significantly improved to include two fundamental additional aspects, i.e., multi-scale decomposition and attention. The model incorporates two independent paths for visual processing corresponding to two different scales. Attention from higher hierarchical levels works by modifying different properties of the network: by selecting the portion of the image to be scrutinized and the appropriate scale, by modulating the threshold of a gating mechanism, and by modifying the width and/or strength of lateral inhibition. Through computer simulations of real complex and noisy black and-white images, we demonstrate that appropriate selection of the above factors allows accurate analysis of image contours at different levels, from global perception of the overall objects without details, down to a fine examination of minute particulars (such as the lips in a face or the fingers of a hand). Attentive reconfiguration of lateral inhibition plays a key role in the analysis of images at different detail levels. PMID- 18406106 TI - EBV infection renders B cells resistant to growth inhibition via adenylyl cyclase. AB - Cyclic AMP (cAMP) is an important physiological growth inhibitor of lymphoid cells, and the cAMP/protein kinase A (PKA) pathway is disrupted in several immunological disorders and cancers. Epstein Barr virus (EBV) infection of B lymphocytes is responsible for the development of lymphoproliferative disease as well as certain B-lymphoid malignancies. Here we hypothesized that EBV infection might render B lymphocytes resistant to cAMP/PKA-mediated growth inhibition. To test this, we assessed the growth-inhibitory response of cAMP-elevating compounds such as forskolin and isoproterenol, as well as the PKA activator 8-CPT-cAMP in normal B lymphocytes, EBV-infected B cells and in the EBV-negative B lymphoid cell line Reh. We could demonstrate that EBV infection indeed abolished cAMP mediated growth inhibition of B cells. The defect was pinpointed to defective adenylyl cyclase (AC) activation by forskolin and isoproterenol, resulting in reduced formation of cAMP and lack of PKA activation and CREB phosphorylation. In contrast, 8-CPT-cAMP which directly activates PKA was able to inhibit EBV infected B cell growth. The physiological implications of these results were underlined by the observation that the ability of forskolin to inhibit camptothecin-induced apoptosis was abolished in EBV-infected B cells. We conclude that EBV infection of B cells abrogates the activation of AC and thereby cAMP formation, and that this dysfunction renders the cells resistant to growth inhibition via the cAMP/PKA pathway. PMID- 18406107 TI - Assessment of mandibular invasion of squamous cell carcinoma by various modes of imaging: constructing a diagnostic algorithm. AB - The aim of this study was to determine the optimal combination of preoperative examination methods to predict mandibular invasion by squamous cell carcinoma of the oral cavity. Data were gathered prospectively but evaluated retrospectively. The preoperative results of clinical examination, conventional radiography, bone single photon emission computed tomography (SPECT), computed tomography and magnetic resonance imaging were compared to the histological results of resection specimens from 67 patients with tumours, adjacent or fixed to the mandible, histologically confirmed as squamous cell carcinoma. The examination methods with acceptable sensitivity and specificity were selected and diagnostic algorithms were constructed using all possible combinations. The preferred diagnostic algorithm was found to be either computed tomography or magnetic resonance imaging, followed by a bone SPECT in cases where the first scan is negative. A negative bone SPECT rules out mandibular invasion (100% sensitivity). This algorithm accurately predicted mandibular invasion in 85% of the patients, without yielding false negative results. In this study group, application of such an algorithm would have resulted in a reduction of the number of unnecessary mandibular resections by 50%. The suggested, preferred, diagnostic algorithm shortens the preoperative screening process, avoiding unnecessary costs, as well as considerably reducing the number of unnecessary mandibular resections. PMID- 18406108 TI - Involvement of septal Cdk5 in the emergence of excessive anxiety induced by stress. AB - The aim of the present study was to evaluate whether the activation of Cdk5, a protein that has been suggested to participate in higher cognitive functions, is required for the onset of a sensitized anxiety-related behavior induced by stress. The exposure to restraint enhanced both Cdk5 expression in certain subareas of the septohippocampal system, principally in the lateral septum (LS) and septal Cdk5 kinase activity in rats. Behaviorally, restrained wild type mice showed a behavior indicative of enhanced anxiety in the elevated plus maze (EPM). In contrast, unstressed mice and stressed knockout mice, which lacked the p35 protein, the natural activator of Cdk5, displayed similar anxiety-like behavior in the EPM. Finally, the intra-LS infusion of olomoucine - a Cdk5 inhibitor - blocked the enhanced anxiety in the EPM induced by prior stress in rats. All these data provide evidence that septal Cdk5 is required in the emergence of a sensitized emotional process induced by stress. PMID- 18406109 TI - Cation dinitrogen complexes [N(2)...X...N(2)]+, X+=H+, Li+, Na+, Be(2+), Mg(2+). AB - The complexes of dinitrogen with five cations (H(+), Li(+), Na(+), Be(2+) and Mg(2+)) up to four N(2) molecules have been calculated at the MP2/6-311++G(d,p) level. Energetic and geometric aspects have been determined together with absolute shieldings (GIAO). The atoms in molecules methodology has been used to analyze energy, charge and volume of these complexes. PMID- 18406110 TI - Lithium ionic jump motion in the fast solid ion conductor Li(5)La(3)Nb(2)O(12). AB - Using (7)Li NMR line-shape analysis, spin-lattice relaxation measurements and stimulated-echo spectroscopy, we investigate the lithium ionic jump motion in the garnet Li(5)La(3)Nb(2)O(12). Results for two samples are compared, which were annealed at 850( composite function)C (GR-850) and at 900( composite function)C (GR-900), respectively. All (7)Li NMR data consistently show that two lithium species with distinguishable dynamical behaviors coexist in each of the samples. While the less mobile species is the majority component in GR-850, the more mobile species is the majority component in GR-900. (7)Li NMR stimulated-echo spectroscopy provides straightforward access to the correlation functions describing the jumps of the respective majority component in both samples. From the temperature-dependent correlation times, we obtain activation energies of 56 and 32kJmol(-1) for GR-850 and GR-900, respectively. For both samples, the correlation functions substantially deviate from simple exponential behavior, indicating a high complexity of the lithium ionic motion in Li(5)La(3)Nb(2)O(12). PMID- 18406111 TI - Improved pulse schemes for separated local field spectroscopy for static and spinning samples. AB - An improved pulse sequence for SLF experiments based on the magic sandwich (MS) scheme for homo-nuclear dipolar decoupling is proposed. The sequence incorporates a double MS, both on I and S spins and has been named as EXE-MS2. The proposed scheme which has a scaling factor of 1 is observed to be free from low intensity artifacts and provides better line-widths particularly for S spins labeled at multiple sites. The pulse sequence which has been applied on static oriented samples incorporates the EXE scheme where direct polarization of the S spin in the B(0) field is utilized in the place of polarization inversion and is observed to perform well without any loss of sensitivity while ensuring considerable reduction in rf power input into the sample. The EXE scheme has also been tested for solid samples under MAS. PMID- 18406112 TI - Biosynthesis of silver nanocrystals by Bacillus licheniformis. AB - The use of microorganisms for the synthesis of nanoparticles is in the limelight of modern nanotechnology. Using the bacterium Bacillus licheniformis, the biosynthesis of silver nanoparticles was investigated. These silver nanoparticles were characterized by means of UV-vis spectroscopy, scanning electron microscopy (SEM), electron diffraction spectroscopy (EDX) and X-ray diffraction (XRD). The nanoparticles exhibited maximum absorbance at 440 nm in UV-vis spectroscopy. The XRD spectrum of silver nanoparticles exhibited 2theta values corresponding to the silver nanocrystal. SEM micrographs revealed the formation of well-dispersed silver nanoparticles of 50 nm, and the presence of silver was confirmed by EDX analysis. PMID- 18406113 TI - Mathematical properties and parameter estimation for transit compartment pharmacodynamic models. AB - One feature of recent research in pharmacodynamic modelling has been the move towards more mechanistically based model structures. However, in all of these models there are common sub-systems, such as feedback loops and time-delays, whose properties and contribution to the model behaviour merit some mathematical analysis. In this paper a common pharmacodynamic model sub-structure is considered: the linear transit compartment. These models have a number of interesting properties as the length of the cascade chain is increased. In the limiting case a pure time-delay is achieved [Milsum, J.H., 1966. Biological Control Systems Analysis. McGraw-Hill Book Company, New York] and the initial behaviour becoming increasingly sensitive to parameter value perturbation. It is also shown that the modelled drug effect is attenuated, though the duration of action is longer. Through this analysis the range of behaviours that such models are capable of reproducing are characterised. The properties of these models and the experimental requirements are discussed in order to highlight how mathematical analysis prior to experimentation can enhance the utility of mathematical modelling. PMID- 18406114 TI - [Regionalization of perinatal care helps to reduce neonatal mortality and morbidity in very preterm infants and requires updated information for caregivers]. AB - In 1997 a large French epidemiological study (Epipage) showed increased mortality and morbidity in Languedoc Roussillon when compared to other regions of France. In order to update information, we set up a regional database about very preterm infants born in Languedoc-Roussillon since 2003. Our objective was to analyze the evolution of mortality and of the morbidity in very preterm infants between 1997 and 2003-2005. METHODS: We analyzed mortality and the morbidity (respiratory, neurological, digestive) of the very preterm infants born alive between 22 and 32 weeks amenorrhea and admitted alive in neonatology included in Epipage study in 1997 and of those included in the regional database in 2003-2005. Between these 2 periods, professional practices were significantly improved as the perinatal network was set up and perinatal care was regionalized. RESULTS: We analyzed the data collected in 3121 subjects of Epipage study and 1111 subjects of the regional database. We observed a significant reduction (P<0.05) of neonatal mortality (8% versus 23%), rate of bronchopulmonary dysplasia (9% versus 19%) and of periventricular leukomalacia (9% versus 18%). During this period, there were significant increases (P<0.05) in the rates of antenatal corticotherapy (87% versus 61%) and caesarean section (72% versus 38%). CONCLUSION: We observed a significant improvement of morbidity of very preterm infants and a decreased mortality for the youngest subjects which was concomitant of an improvement of the professional's practices. It is necessary to take into account these results to propose relevant informations to the professionals with and thus indirectly to the parents. PMID- 18406115 TI - UK Second National Colorectal Cancer Consensus Meeting 2007. PMID- 18406116 TI - Direct compression properties of chitin and chitosan. AB - Deformation and compaction properties of native amino poly-saccharides chitin and chitosan were studied and compared with those obtained with established pharmaceutical direct compression excipients. An instrumented single-punch tablet machine was used for tablet compaction. The following compression parameters were evaluated: a ratio of crushing strength and compression pressure, plasticity and elasticity factor (PF and EF), tensile strength and R-value. Chitin and chitosan were found to have a marked tendency to plastic deformation, and both showed a good compression behaviour compared with the other direct compression excipients including microcrystalline cellulose. It is concluded that chitin and chitosan are potential co-excipients for direct compression applications. PMID- 18406117 TI - In vivo infrared spectroscopy studies of alkanol effects on human skin. AB - Many studies investigate the permeation of actives through the skin and ignore the role of excipients. The solvents used in formulations will undoubtedly penetrate the skin where they can have a number of effects. They can extract skin lipids, they can alter the fluidity of the lipids and they can alter the polarity of the skin. The degree to which they do this and the depth into the skin where this occurs will depend on the uptake kinetics. The problem is to distinguish the different effects. Using ATR-FTIR and deuterated materials this can be achieved in vivo. The aim of the present study was to study the higher alkanols (hexanol, octanol, decanol) in vivo using a combination of ATR-FTIR spectroscopy and tape stripping. Studies conducted in vivo using deuterated vehicles confirmed the lipid extraction effects of d-hexanol and d-octanol, whereas d-decanol did not change skin lipid content. The uptake of d-decanol was higher than for the other vehicles consistent with previous observations on mouse skin for alkanols of increasing chain length. In general, solvent uptake was proportional to the induced shift in the C-H stretching frequency. Lipid disorder was induced by all vehicles studied in vivo and was proportional to the amount of vehicle present in the skin. PMID- 18406119 TI - The sporadic occurrence of a group I intron-like element in the mtDNA rnl gene of Ophiostoma novo-ulmi subsp. americana. AB - The presence of group I intron-like elements within the U7 region of the mtDNA large ribosomal subunit RNA gene (rnl) was investigated in strains of Ophiostoma novo-ulmi subsp. americana from Canada, Europe and Eurasia, and in selected strains of O. ips, O. minus, O. piceae, O. ulmi, and O. himal-ulmi. This insertion is of interest as it has been linked previously to the generation of plasmid-like mtDNA elements in diseased strains of O. novo-ulmi. Among 197 O. novo-ulmi subsp. americana strains tested, 61 contained a 1.6kb insertion within the rnl-U7 region and DNA sequence analysis suggests the presence of a group I intron (IA1 type) that encodes a potential double motif LAGLIDADG homing endonuclease-like gene (HEG). Phylogenetic analysis of rnl-U7 intron encoded HEG like elements supports the view that double motif HEGs originated from a duplication event of a single-motif HEG followed by a fusion event that combined the two copies into one open reading frame (ORF). The data also show that rnl-U7 intron encoded ORFs belong to a clade that includes ORFs inserted into different types of group I introns, e.g. IB, ID, IC3, IA1, present within a variety of different mtDNA genes, such as the small ribosomal subunit RNA gene (rns), apo cytochrome b gene (cob), NADH dehydrogenase subunit 5 (nad5), cytochrome oxidase subunit 1 gene (coxI), and ATPase subunit 9 gene (atp9). We also compared the occurrence of the rnl-U7 intron in our collection of 227 strains with the presence of the rnl-U11 group I intron and concluded that the U7 intron appears to be an optional element and the U11 intron is probably essential among the strains tested. PMID- 18406118 TI - Characterization of protein factor(s) in rat bronchoalveolar lavage fluid that enhance insulin transport via transcytosis across primary rat alveolar epithelial cell monolayers. AB - The aim of this study was to characterize factor(s) in rat bronchoalveolar lavage fluid (BALF) that enhance(s) insulin transport across primary rat alveolar epithelial cell monolayers (RAECM) in primary culture. BALF was concentrated 7.5 fold using the Centricon device and the retentate was used to characterize the factor(s) involved in enhancing apical-to-basolateral transport of intact 125I insulin across various epithelial cell monolayers. These factor(s) enhanced transport of intact insulin across type II cell-like RAECM (3-fold increase) and type I cell-like RAECM (2-fold increase), but not across Caco-2 or MDCK cell monolayers. The insulin transport-enhancing factor(s) were temperature- and trypsin-sensitive. The mechanism of enhancement did not seem to involve paracellular transport or fluid-phase endocytosis, since fluxes of sodium fluorescein and FITC-dextran (70 kDa) were not affected by the factor(s) in the apical bathing fluid. BALF enhancement of intact 125I-insulin transport was abolished at 4 degrees C and in the presence of monensin, suggesting involvement of transcellular pathways. Sephacryl S-200 purification of BALF retentate, followed by LC-MS/MS, indicated that the high molecular weight (>100 kDa) fractions (which show some homology to alpha-1-inhibitor III, murinoglobulin gamma 2, and pregnancy-zone protein) appear to facilitate transcellular transport of insulin across RAECM. PMID- 18406120 TI - Phylogenetic reassessment of the Teloschistaceae (lichen-forming Ascomycota, Lecanoromycetes). AB - The Teloschistaceae is a widespread family with considerable morphological and ecological heterogeneity across genera and species groups. In order to provide a comprehensive molecular phylogeny for this family, phylogenetic analyses were carried out on sequences from the nuclear ribosomal ITS region obtained from 114 individuals that represent virtually all main lineages of Teloschistaceae. Our study confirmed the polyphyly of Caloplaca, Fulgensia and Xanthoria, and revealed that Teloschistes is probably non-monophyletic. We also confirm here that species traditionally included in Caloplaca subgenus Gasparrinia do not form a monophyletic entity. Caloplaca aurantia, C. carphinea and C. saxicola s. str. groups were recovered as monophyletic. The subgenera Caloplaca and Pyrenodesmia were also polyphyletic. In the subgenus Caloplaca, the traditionally recognized C. cerina group was recovered as monophyletic. Because this study is based solely on ITS, to maximize taxon sampling, the inclusion of phylogenetic signal from ambiguously aligned regions in MP (recoded INAASE and arc characters) resulted in the most highly supported phylogenetic reconstruction, compared with Bayesian inference restricted to alignable sites. PMID- 18406121 TI - Phylogeny of Peronospora, parasitic on Fabaceae, based on ITS sequences. AB - Species concepts are a notoriously difficult taxonomic problem in plant-parasitic fungal-like organisms such as downy mildews (Peronosporomycetes, Peronosporales). This is particularly evident in the largest downy mildew genus, Peronospora, which contains a number of economically important pathogens. Here, we investigate relationships of Peronospora species infecting Fabaceae (angiosperms, Rosidae) originating from various collections from different species of host plants and from different European locations by molecular phylogenetic analysis of ITS sequences. Molecular trees were inferred with ML, MP and Bayesian methods and rooted with Pseudoperonospora. As in other downy mildew groups, molecular data mainly support the use of narrow species delimitations and host range as a taxonomic marker. Fabaceae parasites appear to be subdivided into a number of lineages displaying a considerable degree of host specialization with respect to host genera, as well as host subgenera or species. The number of repeats of a repetitive part of the ITS1 is, within limits, characteristic of subgroups within the cluster of Trifolium parasites. We reveal new hosts for Peronospora found on the Iberian Peninsula. PMID- 18406122 TI - Co-occurrence pulmonary haemosiderosis with coeliac disease in child. AB - Idiopathic pulmonary haemosiderosis (IPH) is a rare and serious disorder in children of unknown aetiolopathogeny. Association of IPH and coeliac disease (CD) is even rarer. Immunological origin of IPH is now well accepted. We report the case of an 11-year-old female admitted for evaluation of recurrent streaky haemoptysis that had been evolving over the previous 9 months. Physical examination revealed weight loss with normal weight, but there was cutaneous and mucosal pallor due to severe anaemia (haemoglobin 4.6g/dl). The chest X-rays showed unilateral alveolo-intertitial infiltrate. Broncho-alveolar lavage revealed 70% haemosiderin-laden macrophages. The diagnosis of IPH was made. Since severe anaemia is disproportionate to radiologic findings, searching associated CD was performed and then confirmed by biological and histological examinations. A gluten-free diet was initiated. Evolution was favourable. Looking for especially CD in IPH should be systematic, even in the absence of gastrointestinal symptoms. PMID- 18406123 TI - Waste management from pulp and paper production in the European Union. AB - Eleven million tonnes of waste are produced yearly by the European pulp and paper industry, of which 70% originates from the production of deinked recycled paper. Wastes are very diverse in composition and consist of rejects, different types of sludges and ashes in mills having on-site incineration treatment. The production of pulp and paper from virgin pulp generates less waste but the waste has similar properties to waste from the production of deinked pulp, although with less inorganics. Due to legislation and increased taxes, landfills are quickly being eliminated as a final destination for wastes in Europe, and incineration with energy recovery is becoming the main waste recovery method. Other options such as pyrolysis, gasification, land spreading, composting and reuse as building material are being applied, although research is still needed for optimization of the processes. Due to the large volumes of waste generated, the high moisture content of the waste and the changing waste composition as a result of process conditions, recovery methods are usually expensive and their environmental impact is still uncertain. For this reason, it is necessary to continue research on different applications of wastes, while taking into account the environmental and economic factors of these waste treatments. PMID- 18406124 TI - Polybrominated diphenyl ethers (PBDEs) in leachates from selected landfill sites in South Africa. AB - The last few decades have seen dramatic growth in the scale of production and the use of polybrominated diphenyl ethers (PBDEs) as flame retardants. Consequently, PBDEs such as BDE -28, -47, -66, -71, -75, -77, -85, -99, -100, -119, -138, -153, -154, and -183 have been detected in various environmental matrices. Generally, in South Africa, once the products containing these chemicals have outlived their usefulness, they are discarded into landfill sites. Consequently, the levels of PBDEs in leachates from landfill sites may give an indication of the general exposure and use of these compounds. The present study was aimed at determining the occurrence and concentrations of most common PBDEs in leachates from selected landfill sites. The extraction capacities of the solvents were also tested. Spiked landfill leachate samples were used for the recovery tests. Separation and determination of the PBDE congeners were carried out with a gas chromatograph equipped with Ni63 electron capture detector. The mean percentage recoveries ranged from 63% to 108% (n=3) for landfill leachate samples with petroleum ether giving the highest percentage extraction. The mean concentrations of PBDEs obtained ranged from ND to 2670pgl(-1), ND to 6638pgl(-1), ND to 7230pgl(-1), 41 to 4009pgl(-1), 90 to 9793pgl(-1) for the Garankuwa, Hatherly, Kwaggarsrand, Soshanguve and Temba landfill sites, respectively. Also BDE -28, -47, -71 and BDE 77 were detected in the leachate samples from all the landfill sites; and all the congeners were detected in two of the oldest landfill sites. The peak concentrations were recorded for BDE-47 at three sites and BDE-71 and BDE-75 at two sites. The highest concentration, 9793+/-1.5pgl(-1), was obtained for the Temba landfill site with the highest BOD value. This may suggest some influence of organics on the level of PBDEs. Considering the leaching characteristics of brominated flame retardants, there is a high possibility that with time these compounds may infiltrate into the groundwater around the sites since most of the sites are not adequately lined. PMID- 18406125 TI - Composting clam processing wastes in a laboratory- and pilot-scale in-vessel system. AB - Waste materials from the clam processing industry (offal, shells) have several special characteristics such as a high salinity level, a high nitrogen content, and a low C/N ratio. The traditional disposal of clam waste through landfilling is facing the challenges of limited land available, increasing tipping fees, and strict environmental and regulatory scrutiny. The aim of this work is to investigate the performance of in-vessel composting as an alternative for landfill application of these materials. Experiments were performed in both laboratory-scale (5L) and pilot-scale (120L) reactors, with woodchips as the bulking agent. In the laboratory-scale composting test, the clam waste and woodchips were mixed in ratios from 1:0.5 to 1:3 (w/w, wet weight). The high ratios resulted in a better temperature performance, a higher electrical conductivity, and a higher ash content than the low-ratio composting. The C/N ratio of the composts was in the range of 9:1-18:1. In the pilot-scale composting test, a 1:1 ratio of clam waste to woodchips was used. The temperature profile during the composting process met the US Environmental Protection Agency sanitary requirement. The final cured compost had a C/N ratio of 14.6, with an ash content of 167.0+/-14.1g/kg dry matter. In addition to the major nutrients (carbon, nitrogen, calcium, magnesium, phosphorus, potassium, sulfur, and sodium), the compost also contained trace amounts of zinc, manganese, copper, and boron, indicating that the material can be used as a good resource for plant nutrients. PMID- 18406126 TI - Achieving "Final Storage Quality" of municipal solid waste in pilot scale bioreactor landfills. AB - Entombed waste in current sanitary landfills will generate biogas and leachate when physical barriers fail in the future, allowing the intrusion of moisture into the waste mass contradicting the precepts of the sustainability concept. Bioreactor landfills are suggested as a sustainable option to achieve Final Storage Quality (FSQ) status of waste residues; however, it is not clear what characteristics the residues should have in order to stop operation and after care monitoring schemes. An experiment was conducted to determine the feasibility to achieve FSQ status (Waste Acceptance Criteria of the European Landfill Directive) of residues in a pilot scale bioreactor landfill. The results of the leaching test were very encouraging due to their proximity to achieve the proposed stringent FSQ criterion after 2 years of operation. Furthermore, residues have the same characteristics of alternative waste stabilisation parameters (low BMP, BOD/COD ratio, VS content, SO4(2-)/Cl- ratio) established by other researchers. Mass balances showed that the bioreactor landfill simulator was capable of practically achieving biological stabilisation after 2 years of operation, while releasing approximately 45% of the total available (organic and inorganic) carbon and nitrogen into the liquid and gas phases. PMID- 18406127 TI - A nanoparticle label/immunochromatographic electrochemical biosensor for rapid and sensitive detection of prostate-specific antigen. AB - We present a nanoparticle (NP) label/immunochromatographic electrochemical biosensor (IEB) for rapid and sensitive detection of prostate-specific antigen (PSA) in human serum. This IEB integrates the immunochromatographic strip with the electrochemical detector for transducing quantitative signals. The NP label, made of CdSe@ZnS, serves as a signal-amplifier vehicle. A sandwich immunoreaction was performed on the immunochromatographic strip. The captured NP labels in the test zone were determined by highly sensitive stripping voltammetric measurement of the dissolved metallic component (cadmium) with a disposable-screen-printed electrode, which is embedded underneath the membrane of the test zone. Several experimental parameters (e.g., immunoreaction time, the amount of anti-PSA-NP conjugations applied) and electrochemical detection conditions (e.g., preconcentration potential and time) were optimized using this biosensor for PSA detection. The analytical performance of this biosensor was evaluated with serum PSA samples according to the "figure-of-merits" (e.g., dynamic range, reproducibility, and detection limit). The results were validated with enzyme linked immunosorbent assay (ELISA) and showed high consistency. It is found that this biosensor is very sensitive with the detection limit of 0.02ngmL(-1) PSA and is quite reproducible (with a relative standard deviation (R.S.D.) of 6.4%). This method is rapid, clinically practical, and less expensive than other diagnostic tools for PSA; therefore, this IEB coupled with a portable electrochemical analyzer shows great promise for simple, sensitive, quantitative point-of-care testing of disease-related protein biomarkers. PMID- 18406128 TI - Electrochemiluminescence of CdSe quantum dots for immunosensing of human prealbumin. AB - We describe a non-labeled electrochemiluminescence (ECL) immunosensor based on CdSe quantum dots (QDs) for the detection of human prealbumin (PAB, antigen). The immunosensor was fabricated by layer by layer coupled with nanoparticle amplification techniques. After two gold nanoparticle layers were self-assembled onto the gold electrode surface through cysteamine, anti-PAB (antibody) were conjugated with -COOH groups of both the CdSe QDs and cysteine, which were linked to the gold nanoparticle-modified electrode. The principle of ECL detection was that the immunocomplex inhibited the ECL reaction between CdSe QDs and K(2)S(2)O(8), which resulted in the decrease of ECL intensity. On the one hand, the immunocomplex increased the steric hindrance. On the other hand, the immunocomplex maybe inhibit the transfer of K(2)S(2)O(8) to the surface of the CdSe QD-electrode. The PAB concentration was determined in the range of 5.0 x 10( 10) to 1.0 x 10(-6) g mL(-1), and the detection limit was 1.0 x 10(-11) g mL(-1). The developed CdSe QD-based ECL immunosensor provides a rapid, simple, and sensitive immunoassay protocol for protein detection, which could be applied in more bioanalytical systems. PMID- 18406129 TI - Nutrients and phytochemicals: from bioavailability to bioefficacy beyond antioxidants. AB - The effect of any dietary compound is influenced by the active bioavailable dose rather than the dose ingested. Depending on the individual predisposition, including genetics and medication, a bioavailable dose may cause different magnitudes of effects in different people. Age might affect the predisposition and thus the requirements for nutrients including phytonutrients (e.g. phytochemicals such as flavonoids, phenolic acids and glucosinolates). These are not essential for growth and development but to maintain body functions and health throughout the adult and later phases of life; they are 'lifespan essentials'. Major mechanisms involved in chronic, age-related diseases include the oxidant/antioxidant balance, but the latest research indicates indirect effects of dietary bioactives in vivo and adaptive responses in addition to direct radical scavenging. PMID- 18406130 TI - Challenges in discovering bioactives for the food industry. AB - Nutritional interventions are associated with transient deviations of homeostasis within the human body. The role of a balanced nutrition is, firstly, to ensure an adequate intake of nutrients to efficiently enable all metabolic processes and, secondly, to contribute to sustained human health. This is a new challenge for the food industry because consumers demand taste and convenience in addition to healthy food. The attempts of the food industry to omit health-adverse compounds, and incorporate constituents with identified beneficial health effects has coined the term 'functional food', encompassing fresh or processed food with health promoting and/or disease-preventing properties. The active ingredients of these products are known as 'nutraceuticals'. This paper outlines scientific concepts applied to the identification of novel bioactive food ingredients. PMID- 18406131 TI - Seeing is believing: engineering anthocyanin and carotenoid biosynthetic pathways. AB - The biosynthetic pathways of flavonoids and carotenoids have been well established, and the biosynthetic genes have been mostly isolated. Metabolic engineering of their biosynthetic pathways has provided not only novel colored or health-beneficial plants but also excellent models to study the efficacy of such engineering. In order to achieve a specific color by accumulating a corresponding compound, it is necessary to upregulate the pathway leading to the compound and downregulate the competing pathway. The regulation of gene expression has to be optimized in a target crop as well. PMID- 18406132 TI - Showering c-MET-dependent cancers with drugs. AB - The receptor tyrosine kinase, c-MET and its ligand hepatocyte growth factor/scatter factor (HGF/SF) have become leading candidates for targeted cancer therapies. Inappropriate c-MET signaling through autocrine, paracrine, amplification, and mutational activation occurs in virtually all types of solid tumors (http://www.vai.org/met), contributing to one or a combination of proliferative, invasive, survival, or angiogenic cancer phenotypes. c-MET and HGF/SF participate in all stages of malignant progression and represent promising drug targets in a variety of cancer types, including carcinomas, sarcomas, and brain tumors. While many are in pre-clinical testing, a few inhibitors have entered clinical trials. With hundreds of thousands of potential responding cancers that express c-MET, the interest in this molecule as a drug target is not surprising. However, the cognate c-MET diagnostic tests lag behind. In addition, despite the great enthusiasm based on response rates in phase I trials, there is a need for caution. It is almost without question that combination therapies with c-MET-HGF/SF inhibitors will be required for most cancers to achieve a cytotoxic tumor response. PMID- 18406134 TI - Macromolecular assemblages. Editorial overview. PMID- 18406135 TI - Design and synthesis of tacrine-ferulic acid hybrids as multi-potent anti Alzheimer drug candidates. AB - Five tacrine-ferulic acid hybrids (6a-e) were designed and synthesized as multi potent anti-Alzheimer drug candidates. All target compounds have better acetylcholinesterase inhibitory activity and comparable butyrylcholinesterase inhibitory activity in relation to tacrine. Interestingly, 6d showed a reversible and non-competitive inhibitory action for acetylcholinesterase indicating interaction with the peripheral anionic site, whereas a reversible but competitive inhibitory action for butyrylcholinesterase. The antioxidant study revealed that four target compounds have, compared to Trolox, high ability to absorb reactive oxygen species. PMID- 18406133 TI - The structural biology of HIV assembly. AB - HIV assembly and replication proceed through the formation of morphologically distinct immature and mature viral capsids that are organized by the Gag polyprotein (immature) and by the fully processed CA protein (mature). The Gag polyprotein is composed of three folded polypeptides (MA, CA, and NC) and three smaller peptides (SP1, SP2, and p6) that function together to coordinate membrane binding and Gag-Gag lattice interactions in immature virions. Following budding, HIV maturation is initiated by proteolytic processing of Gag, which induces conformational changes in the CA domain and results in the assembly of the distinctive conical capsid. Retroviral capsids are organized following the principles of fullerene cones, and the hexagonal CA lattice is stabilized by three distinct interfaces. Recently identified inhibitors of viral maturation act by disrupting the final stage of Gag processing, or by inhibiting the formation of a critical intermolecular CA-CA interface in the mature capsid. Following release into a new host cell, the capsid disassembles and host cell factors can potently restrict this stage of retroviral replication. Here, we review the structures of immature and mature HIV virions, focusing on recent studies that have defined the global organization of the immature Gag lattice, identified sites likely to undergo conformational changes during maturation, revealed the molecular structure of the mature capsid lattice, demonstrated that capsid architectures are conserved, identified the first capsid assembly inhibitors, and begun to uncover the remarkable biology of the mature capsid. PMID- 18406136 TI - Spatial patterning of metabolism by mitochondria, oxygen, and energy sinks in a model cytoplasm. AB - Metabolite gradients might guide mitochondrial localization in cells and angiogenesis in tissues. It is unclear whether they can exist in single cells, because the length scale of most cells is small compared to the expected diffusion times of metabolites. For investigation of metabolic gradients, we need experimental systems in which spatial patterns of metabolism can be systematically measured and manipulated. We used concentrated cytoplasmic extracts from Xenopus eggs as a model cytoplasm, and visualized metabolic gradients formed in response to spatial stimuli. Restriction of oxygen supply to the edge of a drop mimicked distance to the surface of a single cell, or distance from a blood vessel in tissue. We imaged a step-like increase of Nicotinamide adenine dinucleotide (NAD) reduction approximately 600 microm distant from the oxygen source. This oxic-anoxic switch was preceded on the oxic side by a gradual rise of mitochondrial transmembrane potential (Deltapsi) and reactive oxygen species (ROS) production, extending over approximately 600 microm and approximately 300 microm, respectively. Addition of Adenosine triphosphate (ATP) consuming beads mimicked local energy sinks in the cell. We imaged Deltapsi gradients with a decay length of approximately 50-300 microm around these beads, in the first visualization of an energy demand signaling gradient. Our study demonstrates that mitochondria can pattern the cytoplasm over length scales that are suited to convey morphogenetic information in large cells and tissues and provides a versatile model system for probing of the formation and function of metabolic gradients. PMID- 18406137 TI - Ino80 chromatin remodeling complex promotes recovery of stalled replication forks. AB - BACKGROUND: Chromatin remodeling complexes facilitate the access of enzymes that mediate transcription, replication or repair of DNA by modulating nucleosome position and/or composition. Ino80 is the DNA-dependent Snf2-like ATPase subunit of a complex whose nucleosome remodeling activity requires actin-related proteins, Arp4, Arp5 and Arp8, as well as two RuvB-like DNA helicase subunits. Budding yeast mutants deficient for Ino80 function are not only hypersensitive to reagents that induce DNA double-strand breaks, but also to those that impair replication fork progression. RESULTS: To understand why ino80 mutants are sensitive to agents that perturb DNA replication, we used chromatin immunoprecipitation to map the binding sites of the INO80 chromatin-remodeling complex on four budding yeast chromosomes. We found that Ino80 and Arp5 binding sites coincide with origins of DNA replication and tRNA genes. In addition, Ino80 was bound at 67% of the promoters of genes that are sensitive to ino80 mutation. When replication forks were arrested near origins in the presence of hydroxyurea (HU), the amount of INO80 complex at stalled forks and at unfired origins increased selectively. Importantly, the resumption of DNA replication after release from a HU block was impaired in ino80 mutants. These cells accumulated double-strand breaks as they attempted to restart replication. Consistently, ino80-deficient cells, although proficient for checkpoint activation, delay recovery from the checkpoint response. CONCLUSIONS: The INO80 chromatin remodeling complex is enriched at stalled replication forks, where it promotes the resumption of replication upon recovery from fork arrest. PMID- 18406138 TI - Comparison of upper extremity kinematics in children with spastic diplegic cerebral palsy using anterior and posterior walkers. AB - This prospective study analyzes the upper extremity kinematics of 10 children with spastic diplegic cerebral palsy using anterior and posterior walkers. Although both types of walkers are commonly prescribed by clinicians, no quantitative data comparing the two in regards to upper extremity motion has been published. The study methodology included testing of each subject with both types of walkers in a motion analysis laboratory after an acclimation period of at least 1 month. Overall results showed that statistically, both walkers are relatively similar. With both anterior and posterior walkers, the shoulders were extended, elbows flexed, and wrists extended. Energy expenditure, walking speed and stride length was also similar with both walker types. Several differences were also noted although not statistically significant. Anterior torso tilt was reduced with the posterior walker and shoulder extension and elbow flexion were increased. Outcomes analysis indicated that differences in upper extremity torso and joint motion were not dependent on spasticity or hand dominance. These findings may help to build an understanding of upper extremity motion in walker assisted gait and potentially to improve walker prescription. PMID- 18406139 TI - Escherichia coli and its chromosome. AB - The Escherichia coli chromosome is a circular DNA molecule that is approximately 1000 times compacted in the living cell, where it occupies approximately 15% of the cellular volume. The genome is organized in a way that facilitates chromosome maintenance and processing. Despite huge efforts, until recently little has been known about how the chromosome is organized within cells, where replication takes place, and how DNA is segregated before cell division. New techniques for labeling genetic loci and molecular machines are allowing the simultaneous tracking of genetic loci and such machines in living cells over time. These studies reveal remarkable organization, yet a highly dynamic flux of genetic loci and macromolecules. It seems likely that the cellular positioning of chromosomal loci is the outcome of the formation of two chromosome arms (replichores) by replication, followed by sequential chromosome segregation, rather than from the presence of cellular positioning markers. PMID- 18406140 TI - The parvovirus capsid odyssey: from the cell surface to the nucleus. AB - During cellular entry and infection, the parvovirus capsid follows a complex path from the cell surface to the nucleus, where the DNA is replicated. Various receptors have been characterized that bind to different parvoviruses and mediate their entry into cells. However, the subsequent trafficking pathways within the endosomal system, cytoplasm and into the nucleus are still not well defined. Studies of viruses entering various cell types under different conditions show particles located in many different endosomal compartments, within the cytoplasm and in the nucleus with significant variations in timing and distribution. Here, we define the previously unresolved issues that are now better understood for the infection pathways of these viruses, and outline some of the areas that remain to be clarified in future studies. PMID- 18406141 TI - (99m)Tc-Tetrofosmin SPECT for the assessment of meningioma malignancy. PMID- 18406142 TI - Peritumoral edema and prognosis in intracranial meningioma surgery. AB - Peritumoral brain edema (PTBE) is often associated with meningiomas. PTBE is probably implicated in the complications occurring in intracranial meningiomas. The goal of this study was to determine the exact implication of PTBE in prognosis. Thirty consecutive patients who underwent surgery for intracranial meningiomas were investigated over a 1-year period. We focused on the clinical and radiological status before and after surgery, and postoperative complications. Multiple regression analysis revealed a close correlation (p<0.05) between PTBE and symptoms, type of arterial supply, difficulty of surgical removal, and postoperative complications. PTBE is likely implicated in the morbidity of intracranial meningiomas. We suggest predictive factors for difficult surgical resection, and emphasise the importance of medical preoperative management and post-operative follow-up. PMID- 18406143 TI - Straight sinus stenosis as a proposed cause of perimesencephalic non-aneurysmal haemorrhage. AB - Perimesencephalic nonaneurysmal haemorrhage (PMNH), a term first coined in 1985, is a diagnosis of exclusion. It is considered a benign form of subarachnoid haemorrhage with a good prognosis and a low risk of rebleeding. Its precise aetiology has not yet been determined. Haemorrhage of venous origin has been proposed as a cause, and has been demonstrated to be associated with venous abnormalities of the basal vein of Rosenthal. We present a patient with PMNH in whom an 80% stenosis at the union of the vein of Galen with the straight sinus is demonstrated. We thus propose hypertensive venous rupture secondary to venous sinus stenosis as an alternative cause for PMNH. PMID- 18406144 TI - Stereotactic treatment of refractory obsessive compulsive disorder by bilateral capsulotomy with 3 years follow-up. AB - In order to study the clinical effect of bilateral capsulotomy in patients with refractory obsessive compulsive disorder (OCD), 35 patients with refractory obsessive compulsive disorder for whom anti-OCD medications and psychological/behavior therapy had failed, underwent MRI-guided stereotactic bilateral anterior capsulotomy. Pre- and post-operative Yale-Brown obsessive compulsive scale (Y-BOCS), Hamilton depression scale (HAMD) and Hamilton anxiety scale (HAMA) scores were determined by psychiatrists. All patients underwent fluorodeoxyglucose positron emission tomography evaluation before and 6 months after the operation. Twenty patients became OCD symptom-free (57%), 10 experienced significant improvement (29%) and five experienced no significant improvement (14%). There were significant decreases in Y-BOCS, HAMD and HAMA scores. Our results show that MRI-guided stereotactic bilateral capsulotomy is a precise, safe and effective therapy for refractory obsessive compulsive disorder. This promising technique may also improve anxiety and depression in addition to OCD. OCD patients who have not responded to medication, psychotherapy or behavioral therapy might benefit from MRI-guided stereotactic bilateral capsulotomy. PMID- 18406145 TI - Enhanced regeneration in spinal cord injury by concomitant treatment with granulocyte colony-stimulating factor and neuronal stem cells. AB - Granulocyte colony-stimulating factor (G-CSF) inhibits programmed cell death and stimulates neuronal progenitor differentiation. Neuronal stem cells transplanted into injured spinal cord can survive, differentiating into astroglia and oligodendroglia, and supporting axon growth and myelination. Herein, we evaluate the combined effects of G-CSF and neuronal stem cells on spinal cord injury. For 40 Sprague-Dawley rats (n=10 in each group) transverse spinal cord resections at the T8-9 level were carried out, leaving an approximately 2-mm gap between the distal and proximal ends of the cord. Neuronal stem cells embedded in fibrin glue treated with or without G-CSF (50 microg/kg x 5 days) (groups III and IV) or fibrin glue with or without G-CSF (50 microg/kg x 5 days) (groups I and II) were transplanted into the gap in the injured spinal cord. Spinal cord regeneration was assessed using a clinical locomotor rating scale scores and electrophysiological, histological and immunohistochemical analysis 3 months after injury. Regeneration was more advanced in group IV than in groups III or II according to the clinical motor score, motor evoked potential, and conduction latency. Most advanced cord regeneration across the gap was observed in group IV rats. Higher densities of bromodeoxyuridine in the injured area and higher expression levels of Neu-N and MAP-2 over the distal end of the injured spinal cord were observed in group IV compared with groups II or III, but there was no significant difference in expression of glial fibrillary acid protein. This synergy between G-CSF and neuronal stem cells may be due to increased proliferation of progenitor cells in the injured area and increased expression of neuronal stem cell markers extrinsically or intrinsically in the distal end of injured cord. PMID- 18406146 TI - Incidence, morphology and clinical relevance of bony canals and vascular grooves in the paracondylar region of adult human skulls. AB - Most features of the skull are well described in standard anatomy textbooks. There may, however, be variations in the form of minor grooves, canals and foramina that merit documentation. In the present study, morphological diversity in the form of osseous tunnels was seen adjacent to the occipital condyles. These canals were present in the paracondylar region (lateral to the occipital condyle) and are therefore termed paracondylar canals. A total of 304 dry adult human skulls (608 sides) of Indian origin were examined and paracondylar canals were found in 59 of these. Thirty-six (11.8%) were bilateral and 23 (7.6%) unilateral. Vascular grooves were found leading to the canals in 52 sides (15.8%). This may result from normal developmental processes and could be a peculiarity confined to the Indian population, as it has not been reported previously. This variant may serve as a valuable anthropological marker. The results of surgical procedures in this area, such as the paracondylar approach (which is directed through the area lateral to the occipital condyles) and the far lateral supracondylar approach, could be impacted upon by this variation. PMID- 18406147 TI - Successful treatment of a blood blister-like aneurysm of the internal carotid artery by trapping with a high-flow bypass. AB - Treatment of blood blister-like aneurysms of the internal carotid artery (ICA) is difficult because the wall of the aneurysm is fragile and there is a high risk of rebleeding. There has been no consensus on the best way to treat these aneurysms. A 32-year-old woman presented with subarachnoid hemorrhage (SAH) caused by a ruptured blood blister-like aneurysm of the ICA. The site of the aneursym was clipped. Although angiography 1week after the operation showed that the aneurysm had been treated successfully, 3 weeks after the initial operation, the aneurysm was found to have recurred to the distal side. In a second operation, the aneurysm was successfully treated by trapping with a high-flow bypass. This case shows that clipping of the rupture site can be insufficient to treat blood blister-like aneurysm of the ICA causing SAH, even if the aneurysm seems to have resolved in follow-up angiographic studies. Trapping of the ICA with a bypass, if necessary a high-flow bypass, is recommended. PMID- 18406148 TI - Choose your partners: dimerization in eukaryotic transcription factors. AB - In many eukaryotic transcription factor gene families, proteins require a physical interaction with an identical molecule or with another molecule within the same family to form a functional dimer and bind DNA. Depending on the choice of partner and the cellular context, each dimer triggers a sequence of regulatory events that lead to a particular cellular fate, for example, proliferation or differentiation. Recent syntheses of genomic and functional data reveal that partner choice is not random; instead, dimerization specificities, which are strongly linked to the evolution of the protein family, apply. Our focus is on understanding these interaction specificities, their functional consequences and how they evolved. This knowledge is essential for understanding gene regulation and designing a new generation of drugs. PMID- 18406149 TI - It takes a PHD to SUMO. AB - PHD fingers and bromodomains are found in close proximity to each other in many chromatin-associated proteins and can functionally synergize. Recently, it has been demonstrated that the PHD finger of the KAP1 co-repressor functions as an E3 SUMO ligase for the adjacent bromodomain. This PHD-mediated SUMOylation stabilizes the association of the bromodomain with the chromatin modifiers SETDB1 and the nucleosome remodeling and deacetylation (NuRD) complex, thereby promoting establishment of the silent gene expression state. PMID- 18406150 TI - Halogenated derivatives QSAR model using spectral moments to predict haloacetic acids (HAA) mutagenicity. AB - The risk of the presence of haloacetic acids in drinking water as chlorination by products and the shortage of experimental mutagenicity data for most of them requires a research work. This paper describes a QSAR model to predict direct mutagenicity for these chemicals. The model, able to describe more than 90% of the variance in the experimental activity, was developed with the use of the spectral moment descriptors. The model, using these descriptors with multiplicative effects provides better results than other linear descriptors models based on Geometrical, RDF, WHIM, eigenvalue-based indices, 2D autocorrelation ones, and information descriptors, taking into account the statistical parameters of the model and the cross-validation results. The structural alerts and the mutagenicity-predicted values from the model output are in agreement with references from other authors. The mutagenicity predicted values for the three haloacetic acids, which have available experimental data (TCAA-Trichloroacetic acid, BDCAA-Bromodichloroacetic acid, and TBAA Tribromoacetic acid), are reasonably close to their experimental values, specially for the latest two. PMID- 18406151 TI - Antiglioma action of xanthones from Gentiana kochiana: Mechanistic and structure activity requirements. AB - The present study identifies xanthones gentiakochianin and gentiacaulein as the active principles responsible for the in vitro antiglioma action of ether and methanolic extracts of the plant Gentiana kochiana. Gentiakochianin and gentiacaulein induced cell cycle arrest in G(2)/M and G(0)/G(1) phases, respectively, in both C6 rat glioma and U251 human glioma cell lines. The more efficient antiproliferative action of gentiakochianin was associated with its ability to induce microtubule stabilization in a cell-free assay. Both the xanthones reduced mitochondrial membrane potential and increased the production of reactive oxygen species in glioma cells, but only the effects of gentiakochianin were pronounced enough to cause caspase activation and subsequent apoptotic cell death. The assessment of structure-activity relationship in a series of structurally related xanthones from G. kochiana and Gentianella austriaca revealed dihydroxylation at positions 7, 8 of the xanthonic nucleus as the key structural feature responsible for the ability of gentiakochianin to induce microtubule-associated G(2)/M cell block and apoptotic cell death in glioma cells. PMID- 18406152 TI - Design, synthesis, and biological evaluation of glycine-based molecular tongs as inhibitors of Abeta1-40 aggregation in vitro. AB - A series of N-terminus benzamides of glycine-based symmetric peptides, linked to m-xylylenediamine and 3,4'-oxydianiline spacers, were prepared and tested as inhibitors of beta-amyloid peptide Abeta(1-40) aggregation in vitro. Compounds with good anti-aggregating activity were detected. Polyphenolic amides showed the highest anti-aggregating activity, with IC(50) values in the micromolar range. Structure-activity relationships suggested that pi-pi stacking and hydrogen bonding interactions play a key role in the inhibition of Abeta(1-40) self assembly leading to amyloid fibrils. PMID- 18406153 TI - Prediction of the aqueous solvation free energy of organic compounds by using autocorrelation of molecular electrostatic potential surface properties combined with response surface analysis. AB - Several quantitative structure-property relationship (QSPR) approaches have been explored for the prediction of aqueous solubility or aqueous solvation free energies, DeltaG(sol), as crucial parameter affecting the pharmacokinetic profile and toxicity of chemical compounds. It is mostly accepted that aqueous solvation free energies can be expressed quantitatively in terms of properties of the molecular surface electrostatic potentials of the solutes. In the present study we have introduced autocorrelation molecular electrostatic potential (autoMEP) vectors in combination with nonlinear response surface analysis (RSA) as alternative 3D-QSPR strategy to evaluate the aqueous solvation free energy of organic compounds. A robust QSPR model (r(cv)=0.93) has been obtained by using a collection of 248 organic chemicals. An external test set based on 23 molecules confirmed the good predictivity of the autoMEP/RSA model suggesting its further applicability in the in silico prediction of water solubility of large organic compound libraries. PMID- 18406154 TI - New R/S-3,4-dihydro-2,2-dimethyl-6-halo-4-(phenylaminothiocarbonylamino)-2H-1 benzopyrans structurally related to (+/-)-cromakalim as tissue-selective pancreatic beta-cell K(ATP) channel openers. AB - The present work was aimed at exploring a series of R/S-3,4-dihydro-2,2-dimethyl 6-halo-4-(phenylaminothiocarbonylamino)-2H-1-benzopyrans structurally related to (+/-)-cromakalim and differently substituted at the 4- and 6-positions. The biological effects of these putative activators of ATP-sensitive potassium channels (K(ATP)) were characterized in vitro on the pancreatic endocrine tissue (inhibition of insulin release) and on the vascular smooth muscle tissue (relaxation of aorta rings). The biological activity of these new dimethylchroman derivatives was further compared to that of (+/-)-cromakalim, (+/-)-pinacidil, diazoxide and BPDZ 73. Structure-activity relationships indicated that an improved potency for the pancreatic tissue was obtained by introducing a meta- or a para-electron-withdrawing group such as a chlorine atom on the C-4 phenyl ring, independently of the nature of the halogen atom at the 6-position of the benzopyran nucleus. Most original dimethylchroman thioureas were more potent than their 'urea' homologues and even more potent than diazoxide at inhibiting insulin release. Moreover, and unlike (+/-)-cromakalim or (+/-)-pinacidil, such compounds appeared to be highly selective towards the pancreatic tissue. Radioisotopic and fluorimetric investigations indicated that the new drugs activated pancreatic K(ATP) channels. Lastly, conformational studies suggested that the urea/thiourea dimethylchromans can be regarded as hybrid compounds between cromakalim and pinacidil. PMID- 18406155 TI - SAR and QSAR study on 2-aminothiazole derivatives, modulators of transcriptional repression in Huntington's disease. AB - REST/NRSF is a multifunctional transcription factor that represses or silences many neuron-specific genes in both neural and non-neural cells by recruitment to its cognate RE1/NRSE regulatory sites. An increase in RE1/NRSE genomic binding is found in Huntington's disease (HD), resulting in the repression of REST/NRSF regulated gene transcription, among which BDNF, thus representing one of the possible detrimental effectors in HD. Three 2-aminothiazole derivatives were recently identified as potent modulators of the RE1/NRSE silencing activity through a cell-based gene reporter assay. In this study, the structure-activity relationships (SAR) of a library of commercially available 2-aminoisothiazoles diversely substituted at the amino group or at position 4 has been evaluated. A quantitative structure-activity relationship analysis performed using the Phase strategy yielded highly predictive 3D-QSAR pharmacophore model for in silico drug screening. PMID- 18406156 TI - Cloning and characterization of a novel LpWRKY1 transcription factor in tomato. AB - The initiation of defence responses in plants is accompanied by fundamental changes in gene expression: the expression of pathogenesis-related genes is co ordinately regulated with metabolic changes such as down regulation of photosynthesis and induction of sink metabolism. To identify candidate regulators of this co-ordinated regulatory mechanism, the role of WRKY transcription factors in the initiation of defence response was analysed in tomato. A WRKY-type transcription factor (LpWRKY1) from tomato was cloned by a reverse Northern approach. The corresponding mRNA is rapidly and transiently induced after challenging the cells with an elicitor-preparation derived from the wilt inducing fungus Fusarium oxysporum lycopersici (E-FOL) and the fungal elicitor chitosan, whereas the endogenous signals systemin and salicylic acid are inactive. Inhibition of protein biosynthesis by cycloheximide results in sustained induction of mRNA for LpWRKY1. In contrast, the transient induction of the gene encoding LpWRKY1 in response to elicitation by E-FOL is inhibited by the protein kinase inhibitor staurosporine and may be mimicked by the phosphatase inhibitors endothall and cantharidine indicating the involvement of protein phosphorylation in the regulation of WRKY-type transcription factors. Direct proof of this postranslational modification of LpWRKY1 was obtained by demonstrating in-gel kinase assays using recombinant LpWRKY1 as substrate. A 44kDa and a 67 kDa protein kinase were shown to be transiently activated to phosphorylate LpWRKY1 protein in response to elicitation with E-FOL. PMID- 18406157 TI - Identification of genes expressed during early somatic embryogenesis in Pinus radiata. AB - Analysis of cDNA-AFLPs was used to study gene expression underlying the early embryogenic process in the gymnosperm Pinus radiata. Somatic embryogenesis in this species was used as a model as it resulted in the generation of a large number of embryos at defined stages of development. The gene expression patterns of three embryogenic stages were compared with non-embryogenic cells. Fifty transcript-derived fragments (TDFs) that are upregulated and 32 TDFs that are down-regulated in the embryogenic stages were selected, sequenced and their homologies sought in the databases. Expression of a selected subset of differentially expressed genes was confirmed by RT-PCR and their levels of expression were quantified. Of the 50 up-regulated TDFs, 16 are homologous to genes encoding either known or putative proteins in higher plants, 19 are homologous to conifer ESTs and 15 did not show significant matches. Of the down regulated TDFs, 8 are homologous to genes encoding either known or putative proteins, 20 are homologous to conifer ESTs and 4 of them did not show significant matches in DNA or protein sequence database. The known up-regulated genes were similar to genes involved in cellular metabolism and in the stress response and the known down-regulated genes were similar to genes involved in proteolysis, cell wall modification and signaling pathways. Their putative individual function is briefly reviewed based on published information, and the potential roles of these genes in embryo development are discussed. PMID- 18406158 TI - Long-term results of autologous submandibular gland transfer for the surgical treatment of severe keratoconjunctivitis sicca. AB - AIM: The aim of this study was to assess the long-term results of autologous submandibular gland transfer for surgical correction of severe keratoconjunctivitis sicca. PATIENTS AND METHODS: A survey was undertaken of 32 patients who had undergone submandibular gland transfer (42 glands) and by following up 11 patients (15 glands) for 5-10 years. Subjective benefit was evaluated as well as clinical findings at the ocular surface. The biochemical consistency of the secreted "saliva-tear" was analysed and compared with natural submandibular saliva of a matched control-group. The vitality and function of the transplants was tested by means of sialoscintigraphy. Immunohistochemical investigations were carried out in specimens of submandibular tissue, gained during reduction procedures of the transplants to correct secretory excess. RESULTS: Patient evaluation and clinical assessment revealed a long-lasting subjective benefit in 2/3 of the patients and a stabilisation at the ocular surface in all cases. The secretion remained as highly concentrated submandibular saliva. Glandular vitality and function was shown scintgraphically. Immunohistochemical investigations revealed no progressive atrophy after transplantation, the ability of cell division remained intact and there was still neuronal tissue in all transplants, even several years after transfer. As all transplants responded well to parasympathomimetic drugs, this might be an indication of re-innervation of the gland. PMID- 18406159 TI - Progress of the treatment for extended aortic aneurysms; is the frozen elephant trunk technique the next standard in the treatment of complex aortic disease including the arch? AB - Patients with extensive aortic aneurysms involving the ascending aorta, aortic arch, and the descending aorta are still considered to be a challenge for many cardiovascular surgeons. The introduction of the elephant trunk technique by Borst et al. in 1983 has greatly facilitated surgery on this kind of pathology and this technique has been recognized as a standard modality for treatment of extended aortic aneurysms. As a next step, the frozen elephant trunk technique has been introduced in some institutes in the late 1990s. With this technique, surgery is performed through a median sternotomy, and an endovascular stent-graft is placed into the descending aorta in an antegrade fashion through the opened aortic arch. Then the ascending aorta and the aortic arch are replaced conventionally. The frozen elephant trunk technique enables one-stage repair of extended aortic aneurysms in a certain patient cohort with similar operative mortality as with the conventional elephant trunk technique, in which a second stage operation is a prerequisite. Although the surgical strategy should be adjusted specifically to each patient's individual pathology, the frozen elephant trunk technique may become the next standard treatment for extended aortic aneurysm instead of its conventional variant. PMID- 18406160 TI - Conservative treatment for rupture of thoracic aortic aneurysm. AB - We present a case of an 88-year-old female with a history of tuberculosis, who was transferred to our hospital due to the sudden onset of epigastralgia and back pain. A chest X-ray demonstrated a bilateral shadow of the upper lung, which suggested the history of tuberculosis. A computed tomography scan demonstrated a large amount of hematoma from the neck to mediastinum and leakage of contrast medium around the distal aortic arch. We diagnosed rupture of thoracic aortic aneurysm, and selected conservative treatment. The patient was intubated under sedation and blood pressure was controlled with vasodepressors. The patient was completely off the ventilator after 65 days of disease, and the patient was discharged after 4 months. This is the first successful case of conservative therapy for ruptured thoracic aortic aneurysm. PMID- 18406161 TI - Ventricular septal dissection after patch repair of an infarct-related ventricular septal rupture. PMID- 18406162 TI - Right atrial paraganglioma: an unusual primary cardiac tumour. AB - We present a case of an unusual presentation of a rare primary cardiac tumour. There are no more than 50 previously reported cases of primary cardiac paraganglioma in the world literature and only a small proportion of these involve the right side of the heart. Diagnosis is difficult and surgical management is varied and complex. PMID- 18406163 TI - Management of massive pulmonary embolism using recombinant activated Factor VII. AB - Massive pulmonary embolism with concomitant intracranial haemorrhage is a condition associated with high mortality. Emergency embolectomy is indicated in those cases in which medical treatment is not possible. The case of a 65-year-old woman with massive pulmonary embolism after cranial trauma with intracranial haemorrhage and two cardiac arrests prior to surgery is described. The patient suffered a right ventricular failure requiring 6h of extracorporeal circulation. After surgery a bleeding disorder happened despite conventional treatment. The patient was transferred to the ICU with packing. Despite the recent thrombotic event recombinant activated Factor VII (70 microg/kg) was used and the bleeding controlled. No other complications occurred and she was discharged 1 month later. As far as we know this is the first time that recombinant activated Factor VII has been used after a massive pulmonary embolism. PMID- 18406165 TI - Comparative pharmacokinetics of liquid and lyophilized formulations of IV RhIG immune globulin. AB - To compare the pharmacokinetics, safety, and tolerability of the liquid and lyophilized formulations of Rh(0)(D) immune globulin intravenous (human) (IV RhIG) administered intramuscularly (IM) and intravenously (IV). In 2 randomized, parallel arm, blinded, phase I studies, 142 healthy adult volunteers received a single dose of either the liquid or lyophilized formulation administered IM (300 microg in Study 1; 15 microg/kg in Study 2) or IV (50 microg/kg in Study 1). Pharmacokinetics (area under the curve [AUC}, C(max), t(1/2), T(max)) and safety data were assessed over 84 days. The 2 formulations had comparable pharmacokinetics following both IM and IV administration. The ratios (90% confidence intervals) for AUC and C(max) treatment means were, for most assessments, within the predefined FDA acceptance range of 80%-125%, demonstrating the bioequivalence of the liquid and lyophilized formulations. Both formulations were equally well tolerated. Study results demonstrate comparable safety and pharmacokinetic profiles of the liquid and lyophilized formulations of IV RhIG. These findings suggest that the liquid formulation will be therapeutically equivalent to the lyophilized formulation but in a more convenient ready-to-use dosage form that may also reduce preparation errors. PMID- 18406164 TI - Caenorhabditis elegans nuclear receptors: insights into life traits. AB - Nuclear receptors are a class of hormone-gated transcription factors found in metazoans that regulate global changes in gene expression when bound to their cognate ligands. Despite species diversification, nuclear receptors function similarly across taxa, having fundamental roles in detecting intrinsic and environmental signals, and subsequently in coordinating transcriptional cascades that direct reproduction, development, metabolism and homeostasis. These endocrine receptors function in vivo in part as molecular switches and timers that regulate transcriptional cascades. Several Caenorhabditis elegans nuclear receptors integrate intrinsic and extrinsic signals to regulate the dauer diapause and longevity, molting, and heterochronic circuits of development, and are comparable to similar in vivo endocrine regulated processes in other animals. PMID- 18406166 TI - Inhibition of V3-specific cleavage of recombinant HIV-1 gp120 produced in Chinese hamster ovary cells. AB - Specific proteolytic cleavage of the gp120 subunit of the HIV-1 envelope (Env) glycoprotein in the third variable domain (V3) has previously been reported to occur in several cell lines, including Chinese hamster ovary cells that have been used for production of Env-based HIV vaccine candidates. Here we report that this proteolytic activity on JRCSF gp120 is dependent on cell density, medium conditions, and supernatant concentration. The resulting cleaved polypeptides cannot be separated from intact gp120 by conventional or affinity chromatography under non-reducing conditions. Inhibitor studies reveal that Pefabloc and benzamidine, but not chymostatin, block gp120 cleavage in a dose-dependent fashion, suggesting the presence of a trypsin-like serine protease in CHO-K1 cells. The proteolytic activity is increased with certain types of cell culture growth media. A combination of serum-free OptiMEM media during expression and potent protease inhibitors post-expression can effectively prevent HIV gp120 degradation. The same strategy can be applied to the expression and purification of gp120 of other strains or other forms of envelope-based vaccine candidates containing V3 sequences. PMID- 18406167 TI - Expression, purification and immunodetection of a recombinant fragment (residues 179-281) of the G protein from rabies virus ERA strain. AB - The glycoprotein (G) of rabies virus (RV) is important for virus infectivity and induction of the protective immunity. In this study, the region comprising linear epitopes (residues 179-281, ERA strain), named rGERA179-281, was cloned in frame with a hexahistidine tag coding sequence at its N-terminal end and overexpressed in Escherichia coli Rosetta strain. The expression under transcriptional regulation of T7 promoter yielded insoluble protein aggregates in the form of inclusion bodies. The inclusion bodies were solubilized with 6M guanidine HCl and the protein was purified to homogeneity under denaturing conditions. Mass spectrometry data confirmed the identity of the protein. The purified protein (13.8kDa) showed significant reactivity with antibodies present in a therapeutic human rabies immune globulin (HRIG), as demonstrated by immunoblotting analysis. In addition, by in vitro competitive neutralization assay, rGERA179-281 led to a measurable reduction in the ability of HRIG to neutralize rabies virus. These results, along with the good yield obtained, encourage further studies on the more detailed immunological properties of rGERA179-281, such as the ability to induce rabies virus neutralizing antibodies and the production of anti-G monoclonal antibodies, which together, might be useful for the development of new diagnostic methods. PMID- 18406168 TI - Production of a recombinant industrial protein using barley cell cultures. AB - The use of recombinant DNA-based protein production using genetically modified plants could provide a reproducible, consistent quality, safe, animal-component free, origin-traceable, and cost-effective source for industrial proteins required in large amounts (1000s of metric tons) and at low cost (below US$100/Kg). The aim of this work was to demonstrate the feasibility of using barley suspension cell culture to support timely testing of the genetic constructs and early product characterization to detect for example post translational modifications within the industrial protein caused by the selected recombinant system. For this study the human Collagen I alpha 1 (CIa1) chain gene encoding the complete helical region of CIa1 optimized for monocot expression was fused to its N- and C-terminal telopeptide and to a bacteriophage T4 fibritin foldon peptide encoding sequences. The CIa1 accumulation was targeted to the endoplasmic reticulum (ER) by fusing the CIa1 gene to an ER-directing signal peptide sequence and an ER retention signal HDEL. The construct containing the CIa1 gene was then introduced into immature barley half embryos or barley cells by particle bombardment. Transgenic barley cells resulting from these transformations were grown as suspension cultures in flasks and in a Wave bioreactor producing CIa1 similar to CIa1 purified from the yeast Pichia pastoris based on Western blotting, pepsin resistance, and mass spectroscopy analysis. The barley cell culture derived-CIa1 intracellular accumulation levels ranged from 2 to 9 microg/l illustrating the need for further process improvement in order to use this technology to supply material for product development activities. PMID- 18406169 TI - Enhancing the specificity of the enterokinase cleavage reaction to promote efficient cleavage of a fusion tag. AB - In our work with designed minimalist proteins based on the bZIP motif, we have found our His-tagged proteins to be prone to inclusion body formation and aggregation; we suspect this problem is largely due to the His tag, known to promote aggregation. Using AhR6-C/EBP, a hybrid of the AhR basic region and C/EBP leucine zipper, as representative of our bZIP-like protein family, we attempted removal of the His tag with enterokinase (EK) but obtained the desired cleavage product in very small yield. EK is known for proteolysis at noncanonical sites, and most cleavage occurred at unintended sites. We manipulated experimental conditions to improve specificity of proteolysis and analyzed the cleavage products; no effect was observed after changing pH, temperature, or the amount of EK. We then suspected the accessibility of the EK site was impeded due to protein aggregation. We found that the easily implemented strategy of addition of urea (1 4 M) greatly improved EK cleavage specificity at the canonical site and reduced adventitious cleavage. We believe that this enhancement in specificity is due to a more "open" protein structure, in which the now accessible canonical target can compete effectively with adventitious cleavage sites of related sequence. PMID- 18406171 TI - The long coming of computational structural biology. AB - Fifty years ago, the structures of the alpha-helix, the beta-sheet, the alpha helical coiled coil and the collagen triple helix raised the expectation that protein structure could be understood computationally, using a combination of geometric considerations, model-building and parametric equations. The first crystal structures dispelled this hope, revealing a disconcerting lack of regularity in the folding patterns of proteins. Gradually it became clear that the protein folding problem-namely deducing the structure of a protein from its amino acid sequence-was of exceptional difficulty. Its solution has remained outside our reach to this day and, arguably, it represents the most important unsolved problem in molecular biology. Nevertheless, our ability to understand and predict molecular structure by computational means has made steady progress, suggesting that we will eventually conquer the problem, not by a few heroic insights, but by steady advances in biophysical knowledge, biological databases, software applications and raw computer power. Computational structural biology, whose influence is already pervasive, will come to dominate structural approaches in the next decades. PMID- 18406170 TI - Changes in surface topologies of chondrocytes subjected to mechanical forces: an AFM analysis. AB - The cartilage is composed of chondrocytes embedded in a matrix of collagen fibrils interspersed within a network of proteoglycans and is constantly exposed to biomechanical forces during normal joint movement. Characterization of the surface morphology, cytoskeletal structure, adherance and elastic properties of these mechanosensitive cells are crucial in understanding the effects of mechanical forces around a cell and how a cell responds to changes in its physical environment. In this work, we employed the atomic force microscope (AFM) to image cultured chondrocytes before and after subjecting them to mechanical forces in the presence or absence of interleukin-1beta to mimic inflammatory conditions. Nanoscale imaging and quantitative measurements from AFM data revealed that there are distinct changes in cell-surface topology and cytoskeleton arrangement in the cells following treatment with mechanical forces, IL-1beta or both. Our findings for the first time demonstrate that cultured chondrocytes are amenable to high-resolution AFM imaging and dynamic tensile forces may help overcome the effect of inflammatory factors on chondrocyte response. PMID- 18406172 TI - On the binding of Thioflavin-T to HET-s amyloid fibrils assembled at pH 2. AB - Amyloid fibrils are ordered beta-sheet protein or peptide polymers. The benzothiazole dye Thioflavin-T (ThT) shows a strong increase in fluorescence upon binding to amyloid fibrils and has hence become the most commonly used amyloid specific dye. In spite of this widespread use, the mechanism underlying specific binding and fluorescence enhancement upon interaction with amyloid fibrils remains largely unknown. Recent contradictory reports have proposed radically different modes of binding. We have studied the interaction of ThT with fibrils of the prion forming domain of the fungal HET-s prion protein assembled at pH 2 in order to try to gain some insight into the general mechanism of ThT-binding and fluorescence. We found that ThT does not bind to HET-s(218-289) fibrils as a micelle as previously proposed in the case of insulin fibrils. We have measured binding kinetics, affinity and stoichiometry at pH values above and below the pI of the HET-s(218-289) fibrils and found that binding is dramatically affected by pH and ionic strength. Binding is poor at acidic pH, presumably as a result of repulsive electrostatic interaction between the positively charged ThT molecule and the fibril surface. Finally, we found that ThT acquires chiral properties when it is fibril-bound. These results are discussed in relation to the different ThT-binding modes that have been proposed. PMID- 18406173 TI - Minds, persons, and space: an fMRI investigation into the relational complexity of higher-order intentionality. AB - Mental state reasoning or theory-of-mind has been the subject of a rich body of imaging research. Although such investigations routinely tap a common set of regions, the precise function of each area remains a contentious matter. With the help of functional magnetic resonance imaging (fMRI), we sought to determine which areas are involved when processing mental state or intentional metarepresentations by focusing on the relational aspect of such representations. Using non-intentional relational representations such as spatial relations between persons and between objects as a contrast, the results ascertained the involvement of the precuneus, the temporal poles, and the medial prefrontal cortex in the processing of intentional representations. In contrast, the anterior superior temporal sulcus and the left temporo-parietal junction were implicated when processing representations that refer to the presence of persons in relational contexts in general. The right temporo-parietal junction, however, was specifically activated for persons entering spatial relations. The level of representational complexity, a previously unexplored factor, was also found to modulate the neural response in some brain regions, such as the medial prefrontal cortex and the right temporo-parietal junction. These findings highlight the need to take into account the critical roles played by an extensive network of neural regions during mental state reasoning. PMID- 18406174 TI - What is awareness of emotion and how does it aid rational decision making?: reply to Ellis. PMID- 18406176 TI - Radiographic osteoarthritis at three joint sites and FRZB, LRP5, and LRP6 polymorphisms in two population-based cohorts. AB - OBJECTIVE: To examine the association of genetic variation in key players in the Wnt signaling pathway with aspects of osteoarthritis (OA) in two population-based cohort studies: the Rotterdam Study and the Chingford Study. METHODS: Radiographic OA (ROA) was defined as a Kellgren/Lawrence score (K/L) score > or = 2 for the knee and hip. Total hip replacement (THR) was scored. Hand OA was defined as presence of ROA (K/L > or = 2) in two out of three hand joint groups [distal interphalangeal (DIPs), proximal interphalangeal (PIPs), first carpometacarpal (CMC1)/trapezio-scaphoid joint (TS)] of each hand. The concentration of urinary C-terminal cross-linked telopeptide of type II collagen (CTX-II) was standardized to the total urine creatinine. Genotypes for the amino acid variants, Arg200Trp and Arg324Gly of Frizzled-Related protein gene (FRZB), Ala1330Val of Low-density lipoprotein receptor-related protein 5 (LRP5) and Ile1062Val of Low-density lipoprotein receptor-related protein 6 (LRP6), were obtained using the Taqman allelic discrimination assay. A meta-analysis was performed for the FRZB Arg324Gly polymorphism and hip- and knee-OA using RevMan version 4.3. RESULTS: No consistent associations were observed between the FRZB, LRP5 and LRP6 amino acid variants and radiographic hip-, knee-, or hand-OA or THR, in either study population. While power was limited for most studies to date, a meta-analysis of all published studies regarding the FRZB Arg324Gly polymorphism was performed for hip- and knee-OA separately. This showed no significant associations between the Gly324 allele and risk for hip- or knee OA, although there was large heterogeneity between studies for hip OA in females. CONCLUSION: No association was seen between FRZB, LRP5 and LRP6 variants with radiographic osteoarthritic outcomes in two population-based cohorts. In future studies, increased power and standardization of OA-phenotypes are highly recommended for replication studies and to allow meta-analysis. PMID- 18406175 TI - Association between trait emotional awareness and dorsal anterior cingulate activity during emotion is arousal-dependent. AB - The dorsal anterior cingulate cortex (dACC) is commonly thought to subserve primarily cognitive functions, but has been strongly implicated in the allocation of attention to emotional information. In a previous positron emission tomography (PET) study, we observed that women with higher emotional awareness as measured by the Levels of Emotional Awareness Scale (LEAS) showed greater changes in regional cerebral blood flow (rCBF) in dACC induced by emotional films and recall. In the current study, we tested whether these effects were due to the processing of any non-neutral stimulus, or were specific to conditions of high emotional arousal. Our results extend the previous finding by demonstrating a positive correlation between emotional awareness and dACC activity only in the context of viewing highly arousing pictures. No such relationship was observed when comparing pleasant or unpleasant pictures to neutral or to each other. We also observed that the relationship between LEAS and dACC activity was present in both sexes but stronger in women than men. These results reinforce the concept that greater trait awareness of one's own emotional experiences is associated with greater engagement of the dACC during emotional arousal, which we suggest may reflect greater attentional processing of emotional information. PMID- 18406177 TI - Expression of recombinant anticoagulant hirudin in the differentiated cultures of the porcine mammary epithelial cell line SI-PMEC. AB - To express recombinant proteins in the spontaneously immortalized porcine mammary epithelial cell line (SI-PMEC) currently established in our laboratory, a chemically synthesized DNA fragment encoding the anticoagulant hirudin was used to construct a mammalian expression vector under the control of the goat beta casein regulatory sequence. The vector, named pGB562/Hi, was transfected into the SI-PMEC cells to yield pGB562/Hi/SI-PMEC. The pGB562/Hi/SI-PMEC cells expressed recombinant hirudin only when they were differentiated into functional structures by growth on a Matrigel-coated petri dish supplemented with the lactogenic hormone prolactin. The differentiated pGB562/Hi/SI-PMEC cells produced about 0.5 0.6microg of recombinant hirudin/mg of total cellular protein. These results suggest that the established SI-PMEC cells have pharmaceutical potential to inducibly express bioactive heterogeneous proteins. PMID- 18406178 TI - Odor enrichment increases interneurons responsiveness in spatially defined regions of the olfactory bulb correlated with perception. AB - Odor enrichment enhances rats' ability to discriminate between chemically similar odorants. We show here that this modulation of olfactory perception is accompanied by increases in the density of local inhibitory interneuron expressing Zif268 in response to olfactory stimuli. These changes depend on the overlap of the olfactory bulb activation patterns induced by the enrichment odorants with those induced by the testing odorants, in a manner similar to changes in perception. Moreover, we show that enrichment leads to an alteration of the pattern of Zif268 expression, dependent on the odors used for the enrichment indicating a restructuring of odor representation in the olfactory bulb. PMID- 18406179 TI - Is heparin reversal with protamine after carotid endarterectomy dangerous? AB - OBJECTIVE: Although systemic heparinisation is routine during CEA, reversal with protamine is controversial with 3 studies suggesting increased peri-operative stroke rates and 3 no effect. None included independent peer-review. DESIGN: Non randomised observational study of data derived from a randomised controlled study of anaesthetic technique for CEA. METHODS: Data on heparin and protamine use and risk factors potentially influencing CEA outcome were collected prospectively. Stroke, death, MI, wound haematoma and re-operation rates were recorded following independent peer-review. RESULTS: 1513/2107 patients received heparin alone (H) and 594/2107 had heparin reversed with protamine (H+P). Risk factors for outcome were similar in both groups. The frequency of outcome events (H v H+P) were: stroke: 67/1513 (4.4%) v 17/594 (2.9%), p=0.098; non stroke or MI death: 10/1513 (0.7%) v 5/594 (0.8%), p=0.657; MI: 6/1513 (0.4%) v 3/594 (0.5%), p=0.718; haematoma: 157/1513 (10.4%) v 44/594 (7.4%), p=0.037; re-operation: 51/1380 (3.7%) v 18/565 (3.2%), p=0.581. CONCLUSIONS: These results show a non significant increase in stroke rate in patients receiving heparin alone refuting suggestions that protamine is harmful. Conversely post-operative haematoma was more frequent when protamine was withheld but re-operation rates were no different. Thus protamine use appears safe and should remain a matter for individual surgeon preference. PMID- 18406181 TI - Microsatellites for disentangling underground networks: strain-specific identification of Glomus intraradices, an arbuscular mycorrhizal fungus. AB - The underground network of arbuscular mycorrhizal (AM) fungi is decisive for the above-ground diversity of many plant ecosystems, but tools to investigate the population structure of AM fungi are sorely lacking. Here, we present a bioinformatics approach to identify microsatellite markers in the AM fungus Glomus intraradices. Based on 1958 contigs of this fungus, assembled from public databases, we identified 842 microsatellites. One hundred of them were subjected to closer scrutiny by designing flanking primers and performing an extensive screen to identify polymorphic loci. We obtained 18 polymorphic microsatellite markers, and we found that seven out of eight individual single-spore cultures of G. intraradices could readily be identified by at least five allelic differences, as compared to all other strains. Two single-spore cultures, however, nominally originating from completely different locations, displayed identity at all 18 loci, suggesting with 99.999999% probability that they represent a single clone. PMID- 18406180 TI - Sumoylation and the function of CCAAT enhancer binding protein alpha (C/EBP alpha). AB - CCAAT enhancer binding protein alpha (C/EBP alpha) is the founding member of a family of basic region/leucine zipper (bZIP) transcription factors and is a master regulator of granulopoiesis. It is expressed at high levels throughout myeloid differentiation and binds to the promoters of multiple myeloid-specific genes at different stages of myeloid maturation. Profound hematopoietic abnormalities occur in mice nullizygous for C/EBP alpha including a selective early block in the differentiation of granulocytes. Mutations in C/EBP alpha are present in a subset of patients with AML presenting with a normal karyotype. These mutations can result in the expression of a 30 kDa dominant negative C/EBP alpha isoform, which contributes to loss of C/EBP alpha function. The molecular basis for this observation remains unknown. In addition to phosphorylation, C/EBP alpha is modified, post-translationally by a small ubiquitin-related modifier (SUMO) at a lysine residue (K159), which lies within the growth inhibitory region of the C/EBP alpha protein. Sumoylation at K159 in the C/EBP alpha protein prevents association of the SWI/SNF chromatin remodeling complex with C/EBP alpha, thereby hampering transactivation. In this review, the functional implications of post-translational modification, particularly sumoylation, of C/EBP alpha in normal granulopoiesis and leukemia are considered. PMID- 18406182 TI - A comprehensive study of dog bites in Spain, 1995-2004. AB - Dog bites in humans are a complex problem embracing public health and animal welfare. To prevent dog bites it is necessary to have comprehensive epidemiological data that allow the identification of associated risk patterns. This study was aimed at investigating the problem posed by dog bites in Spain. The epidemiology of medically attended dog bite-related incidents reported in Aragon was analysed from 1995 to 2004. Bite incidents were mostly associated with: (1) low-population areas (71.3/100,000 inhabitants); (2) males and children, particularly those aged 5-9; (3) single injuries directed to the head and neck area in children and to the extremities in adults; (4) young, male, medium to large, owned dogs that were known to the victim; (5) summer months, and (6) specific circumstances such as human interference with knocked down and fighting dogs. In the light of these risk patterns, a wide range of specific preventive measures could be proposed. PMID- 18406183 TI - Compensatory load redistribution in naturally occurring osteoarthritis of the elbow joint and induced weight-bearing lameness of the forelimbs compared with clinically sound dogs. AB - The current study investigated the compensatory load redistribution due to osteoarthritis of the elbow joint using ground reaction forces of all four legs, simultaneously measured on a treadmill with integrated force plates. Three groups of dogs were used: the first group was clinically sound; the second group suffered from a naturally occurring osteoarthritis of the elbow joint, and a reversible lameness was induced in the third group. The naturally occurring osteoarthritis resulted in a compensatory gait pattern to reduce the stress on the affected limb. The load was reduced on the lame limb and increased on the contralateral hindlimb. The symmetry index indicated a weight-shift to the contralateral forelimb and diagonal hindlimb, which resulted in a more balanced weight distribution than in normal dogs. Dogs with induced lameness showed comparable but less pronounced alterations. These results suggested that forelimb lameness could lead to overload on non-affected extremities and the vertebral spine. PMID- 18406184 TI - The pathobiology of exercise-induced superficial digital flexor tendon injury in Thoroughbred racehorses. AB - Despite the high incidence of superficial digital flexor tendon (SDFT) injury in racehorses, the pathobiology of the condition is not clearly defined. The SDFT improves locomotor efficiency by storing elastic energy, but as a result it has low mechanical safety margins. As with the Achilles tendon in humans, rupture during athletic activity often follows accumulation of exercise and age-induced degenerative change that is not repaired by tenocytes. There is limited understanding of tenocyte biology and pathology, including responses to high mechanical strains and core temperatures during exercise. Unfortunately, much of the current information on SDFT pathology is derived from studies of collagenase induced injury, which is a controversial model. Following rupture the overlapping phases of reactive inflammation, proliferation, remodelling and maturation do not necessarily reconstitute normal structure and function, resulting in long-term persistence of scar tissue and high re-injury rates. Tissue engineering approaches are likely to be applicable to SDFT lesions, but will require significant advances in cell biology research. PMID- 18406185 TI - Maroteaux-Lamy syndrome: functional characterization of pathogenic mutations and polymorphisms in the arylsulfatase B gene. AB - Mucopolysaccharidosis VI (MPS VI; Maroteaux-Lamy syndrome) is an autosomal recessive lysosomal disorder caused by deficiency of N-acetylgalactosamine-4 sulfatase (ARSB), which is required for the degradation of dermatan sulfate. We recently reported mutational screening of 12 Spanish and 4 Argentinian MPS VI patients. In the present study, seven missense mutations (c.245T>G [p.L82R], c.413A>G [p.Y138C], c.719C>T [p.S240F], c.922G>A [p.G308R], c.937C>G [p.P313A], c.1340G>T [p.C447F] and c.1415T>C [p.L472P]) were transiently expressed in COS-7 cells and 4-sulfatase activity was measured in cell extracts. All mutations resulted in less than 6% of wild-type enzyme activity, in most cases undetectable. Mutations were expressed in their original haplotype context with respect to two non-synonymous polymorphisms present in the ARSB protein, p.V358M and p.S384N. The three less frequent haplotype combinations yielded an ARSB activity of 16%, 57% and 70%, when compared to the most frequent haplotype (p.358V and p.384S). Western blot analyses showed that the expressed mutations significantly reduced the amount of mature protein. Sub-cellular localization studies of mutant ARSB proteins in fibroblasts of MPS VI patients were performed. RNA analysis confirmed that nonsense-mediated RNA decay had taken place for all mutant alleles (c.1143-1G>C, c.1143-8T>G, p.W322X, c.427delG and c.1142+2T>A) which were candidates for causing RNA degradation by this mechanism. In summary, all the ARSB mutations studied had a significant effect on enzyme activity, protein processing and/or mRNA stability. PMID- 18406186 TI - Effect of deletion of the prostaglandin EP2 receptor on the anabolic response to prostaglandin E2 and a selective EP2 receptor agonist. AB - Studies using prostaglandin E receptor (EP) agonists indicate that prostaglandin (PG) E(2) can have anabolic effects through both EP4 and EP2 receptors. We previously found that the anabolic response to a selective EP4 receptor agonist (EP4A, Ono Pharmaceutical) was substantially greater than to a selective EP2 receptor agonist (EP2A) in cultured murine calvarial osteoblastic cells. To further define the role of the EP2 receptor in PG-mediated effects on bone cells, we examined the effects of EP2A and PGE(2) on both calvarial primary osteoblasts (POB) and marrow stromal cells (MSC) cultured from mice with deletion of one (Het) or both (KO) alleles of the EP2 receptor compared to their wild-type (WT) littermates. Deletion of EP2 receptor was confirmed by quantitative real-time PCR, Western blot and immunohistochemistry. The 1 month-old mice used to provide cells in these studies did not show any significant differences in their femurs by static histomorphometry. EP2A was found to enhance osteoblastic differentiation as measured by alkaline phosphatase mRNA expression and activity as well as osteocalcin mRNA expression and mineralization in the WT cell cultures from both marrow and calvariae. The effects were somewhat diminished in cultures from Het mice and abrogated in cultures from KO mice. PGE(2) effects were greater than those of EP2A, particularly in POB cultures and were only moderately diminished in Het and KO cell cultures. We conclude that activation of the EP2 receptor is able to enhance differentiation of osteoblasts, that EP2A is a true selective agonist for this receptor and that PGE(2) has an additional anabolic effect likely mediated by the EP4 receptor. PMID- 18406188 TI - Screening of diabetic retinopathy: effect of field number and mydriasis on sensitivity and specificity of digital fundus photography. AB - PURPOSE: To evaluate the sensitivity and specificity of one- and three-field, nonmydriatic and mydriatic, and 45 degrees digital colour photography compared with mydriatic indirect ophthalmoscopy for diabetic retinopathy (DR) screening. METHODS: A group of 79 patients (158 eyes) were included in this prospective study. Colour fundus photographs were taken with a Topcon TRC-NW6S digital camera, using four different techniques--single-field nonmydriatic; three-field nonmydriatic; single-field mydriatic; and three-field mydriatic--followed by dilated ophthalmoscopy. Two independent ophthalmologists classified blinded photographs according to the presence or absence of specific diabetic retinal findings. The sensitivity, specificity and agreement (kappa analyses) of the four methods were calculated for the presence or absence of DR and for all diabetic retinal findings. RESULTS: The sensitivity and specificity of digital photography compared with ophthalmoscopy for detection of DR were, respectively: 77 and 99% using single-field nonmydriatic; 92 and 97% using three-field nonmydriatic; 90 and 98% using single-field mydriatic; 97 and 98% using three-field mydriatic. The degrees of agreement for the four methods were 0.82, 0.90, 0.90 and 0.95, respectively. For specific retinal findings, sensitivity was greater for detection of hard exudates, nerve fibre layer haemorrhage and venous beading, and lower for detection of microaneurysms, dot-blot haemorrhage, cotton wool spots and intraretinal microvascular anomalies. CONCLUSION: The three-field strategy without pupil dilation represents a good compromise, with reasonable sensitivity and good comfort (short examination duration, able to drive after photography) favouring patient compliance with the screening programme. PMID- 18406189 TI - Metabolic syndrome and psychosocial deprivation. Data collected from a Paris suburb. AB - AIM: The association between the metabolic syndrome (MetS) and socioeconomic deprivation has already been recognized. The aim of this study was to examine the relationship between MetS and psychosocial deprivation, and to determine whether or not this relationship is independent of gender, age and obesity. SUBJECTS AND METHODS: The sample included 17074 subjects, aged 16-91 years, who attended the Health Examination Centre of the National Health Insurance Service at Bobigny (a northern suburb of Paris). Exclusion criteria were known diabetes and current lipid-lowering treatment. MetS was defined according to NCEP-ATP III criteria. Psychosocial deprivation was analyzed using the Evaluation of Deprivation and Inequalities by Health Examination Centres (EPICES) score, which includes 11 validated questions and ranges from zero to 100. Psychosocial deprivation was defined as a score greater than or equal to 40. RESULTS: The sample included slightly more women (52.5%) than men, and 52.5% were considered to be deprived. These subjects were slightly older (41 vs 40.2 years, P<0.0001). Around half of the study population benefited from social help. MetS prevalence was higher in the deprived group (11.5% vs 9.4%; P<0.0001), and this was confirmed for every age group. Similarly, the prevalence of all MetS components, except for blood glucose, was significantly higher. The deprivation rate increased with the number of MetS criteria in the non-obese. In a multiple logistic regression, female gender (OR=1.50), age (OR>14 for those over 50 compared with those aged 16-19 years) and deprivation (OR=1.22) were independent predictors of MetS. Taking obese and non-obese populations separately, deprivation remained an independent predictor of MetS only in the non-obese (OR=1.30). CONCLUSION: Psychosocial deprivation is an independent determinant of MetS in non-obese people, and elderly psychosocially deprived women, even if not obese, are at a particularly high risk of MetS. Educational efforts need to be developed to prevent MetS and its consequences in this population. PMID- 18406190 TI - An exceptional manifestation of spondylarthropathy: Destructive proximal tibio fibular arthritis. PMID- 18406191 TI - Acute acquired toxoplasmosis presenting as polymyositis and chorioretinitis in immunocompetent patient. AB - The parasite Toxoplasma gondii mainly encysts in brain, retina, myocardium, and skeletal muscle. It has been implicated in the genesis of inflammatory myopathies for years, but the parasite usually cannot be detected in the muscle. It is established, however, that toxoplasmosis can cause myositis either by recent infection or by reactivation. The case of a non-HIV patient who developed an acute polymyositis upon infection by T. gondii is reported. We suggest that all patients with polymyositis should have serological tests for toxoplasmosis as a part of their initial evaluation and early trial of antiprotozoal therapy in case of positive findings. PMID- 18406192 TI - [Obstetrical ultrasonography of tomorrow, where are we going?]. PMID- 18406194 TI - Pressure pulsation in roller pumps: a validated lumped parameter model. AB - During open-heart surgery roller pumps are often used to keep the circulation of blood through the patient body. They present numerous key features, but they suffer from several limitations: (a) they normally deliver uncontrolled pulsatile inlet and outlet pressure; (b) blood damage appears to be more than that encountered with centrifugal pumps. A lumped parameter mathematical model of a roller pump (Sarns 7000, Terumo CVS, Ann Arbor, MI, USA) was developed to dynamically simulate pressures at the pump inlet and outlet in order to clarify the uncontrolled pulsation mechanism. Inlet and outlet pressures obtained by the mathematical model have been compared with those measured in various operating conditions: different rollers' rotating speed, different tube occlusion rates, and different clamping degree at the pump inlet and outlet. Model results agree with measured pressure waveforms, whose oscillations are generated by the tube compression/release mechanism during the rollers' engaging and disengaging phases. Average Euclidean Error (AEE) was 20mmHg and 33mmHg for inlet and outlet pressure estimates, respectively. The normalized AEE never exceeded 0.16. The developed model can be exploited for designing roller pumps with improved performances aimed at reducing the undesired pressure pulsation. PMID- 18406193 TI - VE-cadherin and ACE: markers for sepsis in post mortem examination? AB - Altered expression of endothelial markers - especially adhesion molecules - is diagnostically helpful for diagnosis of ante mortal undiagnosed sepsis. Up to now it is unclear whether (1) expression of Angiotensin converting enzyme (ACE) and/or VE-cadherin (VEC) plays a comparable role, (2) whether expression intensity correlates with post mortem interval. Fifty-nine lung specimens (20 lung specimens with regular morphology from tumour lobectomies, 39 from patients who died of septic ARDS due to microbiologically proven Gram-negative sepsis) were stained with an antibody against ACE (1:80) resp. VEC (1:100). All specimens showed vessel type specific expression patterns for ACE and VEC which was dramatically reduced in sepsis. ACE staining intensity did not correlate with time between death and autopsy. VEC staining was slightly but statistically not significantly reduced with increasing time interval. Pulmonary VEC and ACE expression are reduced in septic ARDS. However, as neither ACE nor VEC expression correlates with time interval between death and post mortem, expression intensities of VEC or ACE are no reliable indices for time elapsed since death. PMID- 18406195 TI - 3D flow study in a mildly stenotic coronary artery phantom using a whole volume PIV method. AB - Blood flow dynamics has an important role in atherosclerosis initiation, progression, plaque rupture and thrombosis eventually causing myocardial infarction. In particular, shear stress is involved in platelet activation, endothelium function and secondary flows have been proposed as possible variables in plaque erosion. In order to investigate these three-dimensional flow characteristics in the context of a mild stenotic coronary artery, a whole volume PIV method has been developed and applied to a scaled-up transparent phantom. Experimental three-dimensional velocity data was processed to estimate the 3D shear stress distributions and secondary flows within the flow volume. The results show that shear stress reaches values out of the normal and atheroprotective range at an early stage of the obstructive pathology and that important secondary flows are also initiated at an early stage of the disease. The results also support the concept of a vena contracta associated with the jet in the context of a coronary artery stenosis with the consequence of higher shear stresses in the post-stenotic region in the blood domain than at the vascular wall. PMID- 18406196 TI - Relationships between density and Young's modulus with microporosity and physico chemical properties of Wistar rat cortical bone from growth to senescence. AB - The aim of this study is to assess density and elastic properties of Wistar rat cortical bone from growth to senescence and to correlate them with morphological and physico-chemical properties of bone. During growth (from 1 to 9 months), bone density and Young's modulus were found to increase from 1659+/-85 to 2083+/-13 kg m(-3) and from 8+/-0.8 to 19.6+/-0.7 GPa respectively. Bone microporosity was found to decrease from 8.1+/-0.7% to 3.3+/-0.7%. Physico-chemical investigations exhibited a mineralization of bone matrix and a maturation of apatite crystals, as protein content decreased from 21.4+/-0.2% to 17.6+/-0.6% and apatite crystal size and carbonate content increased (c-axis length: from 151 to 173 A and CO(3)W%: from 4.1+/-0.3% to 6.1+/-0.2%). At adult age, all properties stabilized. During senescence, a slow decrease of mechanical properties was first observed (from 12 to 18 months, rho=2089+/-14 to 2042+/-30 kg m(-3) and E(3)=19.8 +/-1.3 to 14.8+/-1.5 GPa), followed by a stabilization. Physico-chemical properties stabilized while microporosity increased slightly (from 3.3% to 4%) but not significantly (p>0.05). A multiple regression analysis showed that morphological and physico-chemical properties had significant effects on density regression model. Microporosity had a greater effect on Young's modulus regression model than physico-chemical properties. This study showed that bone structure, mineralization and apatite maturation should be considered to improve the understanding of bone mechanical behaviour. PMID- 18406197 TI - Frequency and treatment of depressive symptoms in a Parkinson's disease registry. AB - Purpose of this cross-sectional study was to estimate the occurrence of depressive symptoms, as related to other clinical data, in a sample of Parkinson's disease (PD) patients (n=226). Furthermore, we examined the medical care of depressive symptoms in this sample. H&Y stages, cognitive status, sleeping disorders, and dysphagia resulted as significant predictors for depression. Prevalence of depressive symptoms was 35.4%. Only 25.0% of patients suffering from moderate to severe depressive symptoms were prescribed antidepressants. This study supports the view that depression may be underrecognized and undertreated in PD patients. A significant proportion of patients continues to experience depressive symptoms despite antidepressive medication. Recognition and treatment of depression remains a challenge for management of PD. Possible coexisting depressive symptoms should be revealed and assessed by standardized interviews in everyday clinical routine. Large scale randomized controlled trials examining efficacy and safety of antidepressants in PD patients are urgently required. PMID- 18406198 TI - Mineralised organic remains from cesspits at the Roman town of Silchester: processes and preservation. AB - Mineralised organic remains (including apple pips and cereal grains) collected during the ongoing excavations of Insula IX at the Roman town of Silchester, Hampshire have been analysed by a combination of SEM-EDX, powder XRD and IR spectroscopy. The experiments included mapping experiments using spatially resolved versions of each technique. IR and powder XRD mapping have been carried out utilising the synchrotron source at The Daresbury Laboratory on stations 11.1 and 9.6. It is concluded that these samples are preserved by rapid mineralisation in the carbonate-substituted calcium phosphate mineral, dahllite. The rapid mineralisation leads to excellent preservation of the samples and a small crystal size. The value of IR spectroscopy in studying materials like this where the crystal size is small is demonstrated. A comparison is made between the excellent preservation seen in this context and the much poorer preservation of mineralised remains seen in Context 5276 or Cesspit 5251. Comments on the possible mechanism of mineralisation of these samples are made. PMID- 18406199 TI - Stopped-flow kinetic fluorimetric studies of the interaction of Ru(II) complex with DNA and its analytical application. AB - A simple, rapid, and sensitive stopped-flow kinetic fluorometric approach was established for the assay of DNA in synthetic samples and real samples by using the measures of initial reaction rate. The increased initial reaction rate is in proportion to the concentration of DNA in the range of 2.0x10(-8)M to 2.1x10( 6)M. The optimum conditions for various parameters on which the binding of Ru(phen)(2)(dppz)(2+) to DNA depends, were investigated. The influence of various surfactants on the interaction was discussed. Furthermore, stopped-flow techniques were employed to determine kinetic parameters of Ru(phen)(2)(dppz)(2+) binding to DNA under pseudo-first-order conditions. It was found that the interaction of Ru(phen)(2)(dppz)(2+) with DNA was very fast. A two-step reaction mechanism, a fast phase followed by a slow phase, was proposed. PMID- 18406201 TI - The use of F-waves as a probe for motor cortex excitability. PMID- 18406202 TI - Somatosensory-evoked fields on magnetoencephalography for epilepsy infants younger than 4 years with total intravenous anesthesia. AB - OBJECTIVE: Patients must remain immobile for magnetoencephalography (MEG) and MRI recordings to allow precise localization of brain function for pre-surgical functional mapping. In young children with epilepsy, this is accomplished with recordings during sleep or with anesthesia. This paper demonstrates that MEG can detect, characterize and localize somatosensory-evoked fields (SEF) in infants younger than 4 years of age with or without total intravenous anesthesia (TIVA). METHODS: We investigated the latency, amplitude, residual error (RE) and location of the N20m of the SEF in 26 infants (mean age=2.6 years). Seventeen patients underwent TIVA and 9 patients were tested while asleep, without TIVA. RESULTS: MEG detected 44 reliable SEFs (77%) in 52 median nerve stimulations. We found 27 reliable SEFs (79%) with TIVA and 13 reliable SEFs (72%) without TIVA. TIVA effects included longer latencies (p<0.001) and lower RE (p<0.05) compared to those without TIVA. Older patients and larger head circumferences also showed significantly shorter latencies (p<0.01). CONCLUSIONS: TIVA resulted in reliable SEFs with lower RE and longer latencies. SIGNIFICANCE: MEG can detect reliable SEFs in infants younger than 4 years old. When infants require TIVA for MEG and MRI acquisition, SEFs can still be reliably observed. PMID- 18406203 TI - Generation of 'positive slow waves' in the preterm EEG: by the brain or by the EEG setup? PMID- 18406204 TI - Effect of biventricular pacing on symptoms and cardiac remodelling in patients with end-stage hypertrophic cardiomyopathy. AB - BACKGROUND: Biventricular (BiV) pacing is an established therapy for heart failure in ischaemic and dilated cardiomyopathy. Its effects in end-stage hypertrophic cardiomyopathy (HCM) are unknown. AIMS: To assess the potential benefits of BiV pacing in patients with symptomatic end-stage HCM. METHODS: Twenty patients with non-obstructive HCM (12 male, mean age 57+/-13 years), left bundle branch block and symptoms of heart failure refractory to medical therapy underwent implantation of a BiV device. NYHA class, echocardiographic parameters and exercise capacity were assessed before and after implantation. RESULTS: At a mean follow-up of 13+/-6 months, an improvement of at least one NYHA class was reported in 8 (40%) patients. A clinical response was associated with an increase in ejection fraction (from 41+/-14% to 50+/-12%, p=0.009), and reductions in left ventricular end-diastolic diameter (from 57+/-6 mm to 52+/-7 mm, p=0.031) and left atrial diameter (from 65+/-8 mm to 57+/-6 mm, p=0.005). Percentage predicted peak oxygen consumption was unchanged in responders but significantly declined in non-responders (p=0.029). CONCLUSIONS: BiV pacing improved heart failure symptoms in a significant proportion of patients with end-stage HCM. Symptomatic improvement was associated with reverse remodelling of the left atrium and ventricle. PMID- 18406205 TI - Positive inotropic effects of epigallocatechin-3-gallate (EGCG) involve activation of Na+/H+ and Na+/Ca2+ exchangers. AB - BACKGROUND: There is evidence that the tea catechin epigallocatechin-3-gallate (EGCG) modulates myocardial contractility. However, the underlying mechanisms remain to be determined. AIMS: To study potential signalling pathways involved in EGCG-induced contractile parameters. METHODS AND RESULTS: EGCG increased fractional shortening in rat cardiac myocytes and enhanced intracellular systolic Ca2+ concentrations. In isolated rat hearts, perfusion with EGCG resulted in significant, dose-dependent increase in peak systolic left ventricular pressure, as well as in contraction and relaxation velocities. Heart rate did not change. Inhibition of the beta1-receptor with metoprolol had no influence on the contractile effects of EGCG. Furthermore, levels of cAMP and phosphorylation of phospholamban did not change with EGCG, indicating that the beta-receptor pathway is not involved. The L-type Ca2+ channel inhibitors, nifedipine and gallopamil, failed to modulate EGCG-induced increase in contractility. However, the myocardial effects and intracellular calcium transients stimulated by EGCG were significantly reduced by the antagonist of the Na+/H+ exchanger (NHE) methyl-N isobutyl amiloride (MIA), and by blocking of the reverse mode of the Na+/Ca2+ exchanger (NCX) by KB-R7943. CONCLUSION: These results indicate that Ca2+ dependent positive inotropic and lusitropic effects of EGCG are mediated in part via activation of the Na+/H+ exchanger and the reverse mode of the Na+/Ca2+ exchanger in the rat myocardium. PMID- 18406206 TI - Morphological variation of Gephyrocapsa oceanica Kamptner 1943 in plankton samples: implications for ecologic and taxonomic interpretations. AB - Morphological analysis of Gephyrocapsa spp. in plankton samples confirms the existence of five out of six morphotypes that were previously reported from Holocene sediments. Our data suggest a much higher diversity within the genus Gephyrocapsa than the currently accepted species circumscriptions. Furthermore, we confirm the morphological species delineations made by Kamptner that allow the separation of three morphological groups within the genus Gephyrocapsa: one group with large bridge angles (G. oceanica var. typica Kamptner 1943), a second group with small bridge angles (G. oceanica var. californiensis Kamptner 1956) and a third group of small coccoliths (G. aperta Kamptner 1963). However, a seemingly continuous transition from small to large coccoliths within G. oceanica var. typica along a temperature gradient points either to a high phenotypic plasticity of G. oceanica var. typica or numerous sibling species highly adapted to specific environmental conditions. Testing of these hypotheses is of utmost importance to understanding the diversity of marine plankton and its evolution, and to assessing the impact of future and past environmental change on primary producers such as coccolithophorids. PMID- 18406207 TI - Morphological and functional characterization of the thoracic portion of blowfly salivary glands. AB - The abdominal portion of the salivary glands in the blowfly has been studied intensively. Here, we examine the thoracic part of the salivary glands, emphasizing structural and functional aspects. The initial segment downstream of the abdominal portion is secretory and resembles the latter in most structural and functional aspects: the apical membrane is enfolded, forms a canalicular system and contains V-H(+)-ATPase that assembles upon stimulation with the hormone serotonin (5-HT); Na,K-ATPase is localized in the basolateral membrane; septate junctions are not prominent, as deduced from immunofluorescence staining for the marker proteins discs large and fasciclin III. 5-HT elicits, at low concentrations, cytoplasmic [Ca2+] oscillations, and, at saturating concentrations, a tonic [Ca2+] rise. The following, so-called "re-absorptive" segment loops through the coiled secretory portion of the salivary gland. The apical membrane of the re-absorptive cells is not enfolded, and septate junctions are prominent. V-H(+)-ATPase and Na,K-ATPase reside on the apical and basolateral membranes, respectively. Finally, re-absorptive cells are also sensitive to 5-HT; however, whereas V-ATPase assembly has a 5-HT concentration dependence similar to other segments, the Ca2+ response occurs only at higher 5-HT concentrations, and displays a different kinetic pattern. PMID- 18406208 TI - Epithelial-mesenchymal transition and the invasive potential of tumors. AB - The development of metastasis requires the movement and invasion of cancer cells from the primary tumor into the surrounding tissue. To acquire such invasive abilities, epithelial cancer cells must undergo several phenotypic changes. Some of these, including alterations in cell adhesion and migration, are reminiscent of those observed during the developmental process termed epithelial-mesenchymal transition (EMT). Several master gene regulatory programs known to promote EMT during development have recently been discovered to play key roles in cancer progression. In particular, the regulation of cell adhesion molecules and the signaling pathways linking them to mechanisms of gene regulation has emerged as an important determinant of tumor cell invasion and metastasis. A deeper understanding of these mechanisms should allow both better diagnosis and the development of specific treatments for invasive cancer. PMID- 18406209 TI - New insight into the role of dendritic cells in malaria immune pathogenesis. AB - The mechanism by which the host develops protective immunity to malaria remains poorly understood. Dendritic cells (DCs) are central to the initiation and regulation of the adaptive immune response. Modulation of DC function might enable Plasmodium to evade the immune system. Millington et al. propose one mechanism by which malaria inhibits DC-T-cell interactions without interfering directly with T-cell receptor engagement. The consequence is a decrease in the co stimulation required to develop an effective immune response. PMID- 18406210 TI - Parental guidance? Trans-generational influences on offspring life history in mosquitoes. AB - Parental effects are important factors that might influence evolutionary and ecological aspects of parasite vectors and the parasites they transmit. A recent study demonstrated the importance of parental rearing conditions on the malaria vector Anopheles stephensi. When parents are reared in a food-limited environment their offspring have increased bloodmeal sizes and larger clutches. The study highlights that ecological studies are vital for understanding vectors of disease and ultimately for developing effective control strategies. PMID- 18406211 TI - Comparative daily profiles with different preparations of lamotrigine: a pilot investigation. AB - Comparative pharmacokinetic data obtained with different preparations of lamotrigine (LTG) are reported for the first time. Nine outpatients reporting problems attributed to shifts in the preparation of LTG used were admitted to hospital. Patients were treated with proprietary LTG for at least 2 weeks prior to admission. Daily profiles (DPs) spanning 24 hours were obtained by blood sampling at 3- or 4-hour intervals on Day 3 after admission. A second DP was obtained under similar conditions after generic LTG therapy for at least 7 days. LTG concentrations were determined by HPLC, and DPs were generated to compare pharmacokinetic parameters. In five of nine patients, parameters deviated beyond +/-10%. Even with the narrower bioequivalence requirements for mandatory substitution in Denmark, there are some patients who experienced serious clinical consequences (relapse in a seizure-free patient, status epilepticus, epidural hematoma due to ataxia with falls) in association with a change in preparation, and significant corresponding alterations in plasma levels could be demonstrated by comparative pharmacokinetic testing. PMID- 18406212 TI - The Brazilian version of the Quality of Life in Epilepsy Inventory for Adolescents: translation, validity, and reliability. AB - OBJECTIVE: The aim of this study was to translate and adapt the Quality of Life in Epilepsy Inventory for Adolescents (QOLIE-AD-48) into Brazilian Portuguese, as well as to evaluate the psychometric properties of this Brazilian version. METHODS: The QOLIE-AD-48 Brazilian version was tested with respect to reliability and validity in 93 adolescents with epilepsy. Patients completed the Self-Esteem Inventory (SEI), Children's Depression Evaluation Scale (CDES), and Adverse Events Profile (AEP). A subset of these patients were reexamined after 3-4 weeks to evaluate the test-retest reliability of the inventory. We assessed validity by comparing the QOLIE-AD-48 Brazilian version with external measures. RESULTS: Internal consistency was satisfactory (Cronbach's alpha=0.87). Test-retest reliability was acceptable as well (intraclass correlation coefficient=0.52 0.81). Moreover, the QOLIE-AD-48 Brazilian version was highly correlated with other scales, demonstrating strong validity. CONCLUSIONS: We conclude that the QOLIE-AD-48 Brazilian version has psychometric properties equivalent to those of the original American-English version and is a valid and reliable instrument for assessment of health-related quality of life in adolescents with epilepsy in Brazil or other Portuguese-speaking countries. PMID- 18406213 TI - Heterogeneous but "standard" coding systems for adverse events: Issues in achieving interoperability between apples and oranges. AB - Monitoring adverse events (AEs) is an important part of clinical research and a crucial target for data standards. The representation of adverse events themselves requires the use of controlled vocabularies with thousands of needed clinical concepts. Several data standards for adverse events currently exist, each with a strong user base. The structure and features of these current adverse event data standards (including terminologies and classifications) are different, so comparisons and evaluations are not straightforward, nor are strategies for their harmonization. Three different data standards - the Medical Dictionary for Regulatory Activities (MedDRA) and the Systematized Nomenclature of Medicine Clinical Terms (SNOMED CT) terminologies, and Common Terminology Criteria for Adverse Events (CTCAE) classification - are explored as candidate representations for AEs. This paper describes the structural features of each coding system, their content and relationship to the Unified Medical Language System (UMLS), and unsettled issues for future interoperability of these standards. PMID- 18406214 TI - Genetic diversity of Echinococcus multilocularis on a local scale. AB - Echinococcusmultilocularis is the causative agent of human Alveolar Echinococcosis (AE), and it is one of the most lethal zoonotic infections in the Northern Hemisphere. In France, the eastern and central regions are endemic areas; Franche-Comte, Lorraine and Auvergne are particularly contaminated. Recently, several human cases were recorded in the French Ardennes area, a region adjacent to the western border of the E. multilocularis range in France. A previous study in this focus described a prevalence of over 50% of the parasite in red foxes. The present study investigated the genetic diversity of adult worms collected from foxes in a 900km(2) area in the Ardennes. Instead of a conventional mitochondrial target (ATP6), two microsatellite targets (EmsB and NAK1) were used. A total of 140 adult worms isolated from 25 red foxes were genotyped. After hierarchical clustering analyses, the EmsB target enabled us to distinguish two main assemblages, each divided into sub-groups, yielding the differentiation of six clusters or assemblage profiles. Thirteen foxes (52% of the foxes) each harbored worms from at least two different assemblage profiles, suggesting they had become infected by several sources. Using the NAK1 target, we identified 3 alleles, two found in association with the two EmsB assemblages. With the NAK1 target, we investigated the parasite breeding system and the possible causes of genetic diversification. Only one fox harbored hybrid worms, indicative of a possible (and rare) occurrence of recombination, although multiple infections have been observed in foxes. These results confirm the usefulness of microsatellite targets for assessing genetic polymorphism in a geographically restricted local range. PMID- 18406215 TI - Base excision repair activities in organotypic hippocampal slice cultures exposed to oxygen and glucose deprivation. AB - The capacity for DNA repair is likely to be one of the factors that determine the vulnerability of neurons to ischemic stress and may influence the pathological outcome of stroke. In this report, initiation of base excision repair (BER) was assessed by analysis of enzyme activity and gene expression level of DNA glycosylases and AP-endonucleases in rat organotypic hippocampal slice cultures exposed to oxygen and glucose deprivation (OGD) - an in vitro model of stroke. Under basal conditions, AP-endonuclease activity and base removal of ethenoadenine and 8-oxoguanine (8-oxoG) were higher (by approximately 20-35 %) in CA3/fascia dentata (FD) than in CA1. Base removal of uracil did not differ between the two hippocampal regions, while removal of 5-hydroxyuracil (5-OHU) was slightly less efficient in CA3/FD than in CA1. Analyses performed immediately after 30 min of OGD revealed a decreased AP-endonuclease activity (by approximately 20%) in CA1 as well as CA3/FD, and an increased ethenoadenine activity (by approximately 25%) in CA1. Activities for 8-oxoG, 5-OHU and uracil showed no significant changes at this time point. At 8h after OGD, none of the enzyme activities differed from control values. Real-time RT-PCR showed that transcription of DNA glycosylases, including Ogg1, Nth1, Ung, Aag, Neil1 and Neil2 were not changed in response to OGD treatment (t=0 h). The hippocampal expression of Neil2 was low compared with the other DNA glycosylases. These data indicate that CA1 has a lower capacity than CA3/FD for removal of base lesions under basal conditions. The relatively low capacity for BER in basal conditions and the apparent failure to upregulate repair of oxidative damage after OGD might contribute to the high vulnerability of CA1 to ischemic injury. PMID- 18406216 TI - TREX1 DNA exonuclease deficiency, accumulation of single stranded DNA and complex human genetic disorders. AB - Aicardi-Goutieres syndrome (AGS) is an unusual condition that clinically mimics a congenital viral infection. Several genes have recently been implicated in the aetiology of this disorder. One of these genes encodes the DNA exonuclease TREX1. Recent work from Yang, Lindahl and Barnes has provided insight into the cellular consequence of TREX1-deficiency. They found that TREX1-deficiency resulted in the intracellular accumulation of single stranded DNA resulting in chronic activation of the DNA damage response network, even in cells from Trex1-mutated AGS patients. Here, I summarise their findings and discuss them in context with the other AGS causative genes which encode subunits of the RNase H2 complex. I describe mechanisms by which the inappropriate intracellular accumulation of nucleic acid species might deleteriously impact upon normal cell cycle progression. Finally, using the example of Systemic Lupus Erythematosus (SLE), I also summarise the evidence suggesting that the failure to process intermediates of nucleic acid metabolism can result in the activation of uncontrolled autoimmunity. PMID- 18406217 TI - VEGF in inflammatory bowel disease: a master regulator of mucosal immune-driven angiogenesis. PMID- 18406218 TI - Gallstones, cholecystectomy and irritable bowel syndrome (IBS) MICOL population based study. AB - BACKGROUND: Subjects with irritable bowel syndrome may undergo an excess of cholecystectomy. It is not known, however, whether the cholecystectomy rate parallels an increased risk of gallstones. AIM: Aim was to assess the prevalence and the incidence of gallstones and cholecystectomy in subjects with irritable bowel syndrome symptoms. SUBJECTS AND METHODS: In this population-based study, 29,139 subjects (63.2% of 46,139 randomly selected subjects, age 30-69 years) underwent a physical examination, an interview on gastrointestinal symptoms and an upper abdominal ultrasonography. An identical survey was carried out 7.8+/-1.0 (M+/-S.D.) years later on 8460 gallstone-free subjects at the first survey. Prevalence and incidence of gallstones and cholecystectomy were assessed in subjects with (1) irritable bowel syndrome; (2) abdominal pain and normal bowel; (3) altered bowel and no abdominal pain and (4) asymptomatic controls; univariate and multivariate regression logistic models were used for statistical analysis. RESULTS: Prevalence odds of gallstones and cholecystectomy were significantly higher in irritable bowel syndrome and abdominal pain and normal bowel than in controls. Irritable bowel syndrome and abdominal pain and normal bowel subjects were more aware of gallstones than controls (p<0.001), and the prevalence of gallstones in irritable bowel syndrome subjects unaware of their gallbladder status was not significantly different from the controls. The incidence of gallstone disease in irritable bowel syndrome, abdominal pain and normal bowel, and altered bowel and no abdominal pain subjects did not differ from the controls. The incidence of cholecystectomy was higher in irritable bowel syndrome and abdominal pain and normal bowel groups than in controls and altered bowel and no abdominal pain group. CONCLUSIONS: Irritable bowel syndrome subjects have an increased risk of cholecystectomy that is not due to an increased risk of gallstones, but rather to abdominal pain, awareness of having gallstones, and inappropriate surgical indications. PMID- 18406219 TI - The impact of biopsy number and site on the accuracy of intestinal metaplasia detection in the stomach A morphometric study based on virtual biopsies. AB - BACKGROUND: Intestinal metaplasia is a risk factor for gastric carcinoma. So far few studies have focused on the efficacy of endoscopic biopsies in detecting intestinal metaplasia in relation with the site and number of biopsies performed. The present study is aimed to assess the efficacy of single and multiple gastric biopsies in the detection and staging of intestinal metaplasia. MATERIAL AND METHODS: The study was conducted on 455 gastrectomies. We defined the extent of intestinal metaplasia on surgical specimens sampled by the "Swiss roll" technique and assessed the efficiency of an increasing number of "virtual" biopsies performed on the same rolls in recognising the true extent of intestinal metaplasia in the antro-angular region. RESULTS: Seventy-four out of 455 cases showed intestinal metaplasia in more than 5% of the antro-angular mucosa. The correlation between antro-angular intestinal metaplasia on rolls and on virtual biopsies was always highly significant, both when a single biopsy was considered and when groups of multiple biopsies (from 2 to 6 samples) were taken into account (por=20% and intestinal metaplasia <20% groups (p<0.0001). The highest value of concordance between intestinal metaplasia extent in virtual biopsies and in rolls was recorded for a set of 4 antral+1 angular biopsies whereas a set of 2 antral+1 angular biopsies turned out to be the most effective in terms of number/efficacy. CONCLUSIONS: A three biopsy set (2 antral+1 angular) allows correct detection of intestinal metaplasia extent in 90% of cases. PMID- 18406220 TI - Successful endoscopic treatment of Bouveret's syndrome by mechanical lithotripsy. AB - Gastric outlet obstruction secondary to the impaction of large biliary stones into the duodenum (Bouveret's syndrome) is a well-known complication of biliary lithiasis, most often requiring surgical intervention. We report a case of successful endoscopic removal of a large stone impacted in the duodenal bulb by means of mechanical lithotripsy. PMID- 18406221 TI - Thin films with chemically graded functionality based on fluorine polymers and stainless steel. AB - Thin films of stainless steel and poly(tetrafluoroethylene) were co-deposited, by radiofrequency magnetron sputtering, in an inert atmosphere in order to produce a functionally graded material as a coating on a traditional biomaterial, where non ferromagnetic characteristics and improved wettability must be ensured. These thin films are intended to modify the surface of SS316L used in stents, where the bulk/thin film couple should be regarded as a single material. This requires excellent adhesion of the coating to the substrate. All coatings were deposited with an average thickness of 500 nm. The chemical and phase characterization of the surface revealed that, with the increase in F content, the thin film evolves from a ferritic phase (alpha) to an amorphous phase with dispersion of a new crystalline ceramic phase (FeF(2)). For intermediate F content values, an austenitic (111) phase (gamma) was present. Bearing in mind the envisaged application, the best results were attained for thin films with a fluorine content between 10 and 20 at.%. PMID- 18406222 TI - Alpha-thalassaemia. AB - Alpha-thalassaemia is one of the most common human genetic disorders. Couples in which both partners carry alpha(0)-thalassaemia traits have a 25% risk of having a fetus affected by homozygous alpha-thalassaemia or haemoglobin Bart's disease, with severe fetal anaemia in utero, hydrops fetalis, stillbirth or early neonatal death, as well as causing various maternal morbidities. This disorder is common in southeast Asia and southern China, and the expanding populations of southeast Asian immigrants in the US, Canada, UK and Europe mean that this disorder is no longer rare in these countries. PMID- 18406223 TI - Comment on "In vivo perfusion analysis of normal and dysplastic ears and its implication on total auricular reconstruction". PMID- 18406224 TI - Reverse epidemiology in hemodialysis patients. Lessons from Japanese registries. PMID- 18406226 TI - Trends in Endocrinology and Metabolism 1998. PMID- 18406225 TI - Delayed pubertal development by hypothalamic suppression causes an increase in periosteal modeling but a reduction in bone strength in growing female rats. AB - The timing of the pubertal growth is a critical event in skeletal development. A delay in the onset of puberty has been correlated with increased stress fracture incidence in young women and as a result, suboptimal skeletal development may affect long-term bone strength. Gonadotropin releasing hormone antagonist (GnRH a) injections were used to delay the onset of puberty in growing female rats. 23 day-old female rats were injected with a GnRH-antagonist at 2 dosage levels (n=15/group). The Low Dose group (1.25 mg/kg/dose) received daily injections for 27 days (sacrifice 49 days). The High Dose group received (5.0 mg/kg/dose) only 5 days per week over a 26 day period (sacrifice 48 days). Calcein injections measured bone formation activity on the periosteal and endocortical surfaces. Standard histomorphometric and biomechanical analyses were performed on the femora and ash content was measured on the tibiae of all animals. Serum estradiol and insulin-like growth factor (IGF)-1 levels were assayed. Significant delays in pubertal development occurred in the two GnRH-a groups as evidenced by delayed vaginal openings, decreased uterine and ovarian weights and suppressed estradiol levels compared to control. Femoral lengths were significantly shorter in the experimental groups and serum IGF-1 levels were higher than control. Bone strength and stiffness were significantly lower in the GnRH-a groups. Cortical bone area was decreased and total area was not different between groups. There was a significant decrease in % Ct.Ar/T.Ar. The decreased bone strength may have resulted from a decrease in the amount and distribution of bone, however, stress and Young's modulus were also decreased. There was a different response between endocortical formation indices and periosteal formation indices to the GnRH-a protocol. Endocortical bone formation rates decreased and there was an increase in periosteal labeled surface. A dose response between bone strength and GnRH-a dosage was found. The data suggest that hypothalamic suppression during pubertal development resulted in decreased bone strength which may result in fracture development. PMID- 18406227 TI - XX germ cells: the difference between an ovary and a testis. AB - In mammals, gonadal sex is determined by the action of the testis-determining gene, SRY. In the absence of SRY, the indifferent gonad follows an alternative pathway and develops as an ovary. Both mitotic and meiotic germ cells appear to play an essential role in ensuring ovarian development. Ovaries depleted of germ cells before or after ovarian differentiation has commenced can develop seminiferous cords, suggesting that XX germ cells may inhibit testicular differentiation in the ovary. PMID- 18406228 TI - Hormonal regulation of osteoclast function. AB - Hormones and cytokines indirectly control the formation of osteoclasts from hemopoietic precursors by acting upon osteoblastic stromal cells and, in some cases, also upon cells of the immune system. These intermediate cells produce factors that act in a paracrine manner to influence precursor proliferation or differentiation. Successful osteoclast formation in vitro requires contact between stromal and hemopoietic cells, leading to the concept of a membrane associated stromal cell molecule that specifically programs osteoclast differentiation. Attention has been focused further on this by the recent discovery of a soluble member of the tumor necrosis factor (TNF) receptor family which is both a product of and a ligand for osteoblastic stromal cells. Once they are formed in the presence of osteoblasts, osteoclasts are active, and hormones or cytokines do not promote the activity of mature osteoclasts, but more likely influence their survival. Of the two best known hormonal inhibitors of bone resorption in vivo, calcitonin acts directly upon osteoclasts to inhibit their activity, whereas estrogen acts indirectly, via the regulation of several cytokines. PMID- 18406229 TI - New aspects of G-protein-coupled receptor signalling and regulation. AB - Research on the structure, regulation and signalling properties of the family of seven-transmembrane-helix, heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors (GPCRs) continues at a frantic pace. This reflects their central role in transmission of hormone- and neurotransmitter-encoded information across the plasma membrane of cells. The location of the ligand binding sites on the extracellular face of the membrane has made them obvious targets for therapeutic intervention in a wide range of conditions resulting from endocrine imbalance. Furthermore, based on the identification of many novel GPCR sequences emerging from expressed sequence tags (ESTs) and other DNA sequencing programmes, it has become clear that the GPCR family is likely to be considerably larger than appreciated in even the recent past. Although neither the natural ligands nor synthetic pharmaceuticals have yet been identified for these so called ;orphan' GPCRs, they offer the potential for a plethora of new therapeutic targets. Within a short review, it is impossible to cover all the current developments in this field and the topics selected represent a personal view of recent highlights of areas that provide both novel and general insights into the function and regulation of GPCRs. PMID- 18406230 TI - Molecular genetics of pituitary tumours. AB - The last several years have seen a significant increase in our understanding of the molecular and biochemical changes associated with pituitary tumour initiation and progression. The combined data, from several groups, now allow a tentative map to be drawn showing that reduction to hemizygosity at several chromosomal loci (10q, 11q13 and 13q) is associated with the transition to the invasive phenotype, while loss on chromosome 9p and methylation of the tumour suppressor gene p16 appear to occur early in pituitary tumorigenesis. Changes in the expression/status of several genes and/or proteins including p53, the cAMP response element-binding factor (CREB), growth hormone-releasing hormone (GHRH), nm23, p16 and p27 have also been identified along this multi-step pathway. Prospective studies will determine whether these markers are truly predictive of subsequent tumour behaviour and can be used to aid clinical management in a manner not possible when current histological criteria are used. PMID- 18406231 TI - Constitutively signaling G-protein-coupled receptors and human disease. AB - Dysregulation of G-protein-coupled receptor (GPCR) function has been shown to be associated with a growing number of human diseases. In some diseases, mutation of an endogenous GPCR causes the receptor to lose the ability to bind agonist or signal (;loss of function' mutation), whereas another mutation causes the receptor to be in an active state in the absence of agonist (;gain of function' mutation), leading to ;constitutive signaling activity'. A number of constitutively active GPCRs are tumorigenic in vitro and in animal models, and cause syndromes of hyperfunction and/or tumors in humans. The recent characterization of a constitutively active GPCR in the genome of a disease associated, human herpesvirus provides a potential novel mechanism for viral tumorigenesis. PMID- 18406238 TI - On the "First International Symposium on the Developmental Aspects of Androgen Excess". PMID- 18406232 TI - Hormone replacement therapy in women with previous breast cancer. AB - As breast cancer is known to be a tumour sensitive to the effects of endogenous oestrogens, clinicians are reluctant to prescribe hormone replacement therapy (HRT) to women with a history of previous breast cancer for fear of stimulating disease recurrence, and it is currently contraindicated in this group of women. However, an increasing proportion of breast cancer patients are requesting the use of HRT to relieve the symptoms of oestrogen deficiency, which are also a common side-effect of adjuvant therapy for breast cancer. Observational data on the use of HRT in breast cancer survivors has not demonstrated an increase in disease recurrence, but uncertainty will continue in the absence of data from prospective, randomized trials. This review aims to demonstrate why it is ethical and scientifically important to undertake such studies. PMID- 18406239 TI - The regulation of human P450c17 activity: relationship to premature adrenarche, insulin resistance and the polycystic ovary syndrome. AB - The polycystic ovary syndrome (PCOS) is characterized by hyperandrogenism and insulin resistance, but the connection between these two features has been unclear. Androgen synthesis is regulated in part by the ratio of the 17alpha hydroxylase and 17,20 lyase activities of P450c17. Three separate lines of evidence show that the ratio of lyase to hydroxylase activity is regulated by electron flow from P450 oxidoreductase. Lyase activity and androgen synthesis are particularly dependent on the serine phosphorylation of P450c17. Serine phosphorylation of the insulin receptor beta chain causes insulin resistance, and some PCOS women have hyperphosphorylated receptors. We hypothesize that an overactive serine/threonine kinase hyperphosphorylates both the insulin receptor and P450c17 in PCOS, accounting for the characteristic insulin resistance and hyperandrogenism of this disease. PMID- 18406240 TI - Current developments in the molecular genetics of the polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is most likely a complex trait with an oligogenic basis. In this article, we present evidence from molecular genetic studies for involvement of the steroid synthesis gene CYP11a (coding for P450 cholesterol side-chain cleavage) in the aetiology of hyperandrogenism. Variation in the regulatory region of the insulin gene also appears to contribute to the development of PCOS. PMID- 18406241 TI - The development of the polycystic ovary syndrome: family history as a risk factor. AB - Three general genetic models for the development of the polycystic ovary syndrome (PCOS) can be proposed, namely: (1) the "single-gene Mendelian" model, which considers the majority of defects present in PCOS to be unique; (2) the "multifactorial" model, which suggests that the defects present in PCOS are not unique, and simply represent the conglomeration of abnormalities already present separately, and to a significant degree, in the general population (e.g. as in cardiovascular disease and non-insulin-dependent diabetes); and (3) the "variable expression-single gene" model, a modified version of the above two. Overall, our data support this third model, suggesting that PCOS is a familial disorder, with a single autosomal dominant gene effect, and a variable phenotype. Family history can then be considered as an important factor determining the risk of developing PCOS. Our preliminary data indicate that a woman's risk of developing PCOS is approximately 40% if her sister is affected. Alternatively, only 19% of mothers were affected, suggesting that the inheritance of PCOS may be preferentially paternal, although expanded clinical studies will be required to confirm these findings. Considering PCOS to be a dominant genetic disorder with a high degree of expressivity, we propose that the risk of developing the disorder is governed by family history and the degree of exposure to the selected environmental and/or other genetic influences. PMID- 18406242 TI - Endocrine and metabolic abnormalities in adolescents with a PCOS-like condition: consequences for adult reproduction. AB - The polycystic ovary syndrome (PCOS) has a perimenarcheal onset. Increased luteinizing hormone (LH) and androgen concentrations are common among anovulatory adolescents and some also have hyperinsulinemia. Many will later have normal ovulatory cycles, whereas others with several co-existing abnormalities will develop full-blown PCOS with menstrual cycle irregularities and infertility in adulthood. PMID- 18406243 TI - Insights into the development of polycystic ovary syndrome (PCOS) from studies of prenatally androgenized female rhesus monkeys. AB - The developmental pathophysiology of polycystic ovary syndrome (PCOS) is unknown. However, prenatally androgenized female rhesus monkeys exhibit ovarian and endocrinological features that mimic those found in women with PCOS. Thus, prenatal androgen excess may provide an etiology for hyperandrogenism and anovulation in adulthood. PMID- 18406244 TI - Markers of abdominal adipose tissue in women: relationship to ovarian function. AB - Increased abdominal adipose tissue (AAT) mass, as most reliably measured by waist circumference, in pubescent girls positively correlates with insulinemia, circulating insulin-like growth factor 1, insulin resistance, androgenemia, and both plasma luteinizing hormone and estradiol levels. Because insulin resistance and androgen excess regularly accompany the ovarian changes seen in polycystic ovary syndrome (PCOS), we propose that elevated AAT mass in adolescent girls, acting through an increase in insulin resistance, might make them more prone to the development of full-blown PCOS. PMID- 18406245 TI - Insulin resistance, premature adrenarche, and a risk of the Polycystic Ovary Syndrome (PCOS). AB - Timing of puberty and final height are usually normal in girls with a history of premature adrenarche. However, these patients show an increased frequency of ovarian hyperandrogenism, hyperinsulinism and dyslipemia at adolescence. The hyperinsulinemia and lipid disturbances can often be detected in the prepubertal period, recommending long-term follow-up of these patients into adulthood. PMID- 18406246 TI - Insulin resistance in prepubertal African-American and Hispanic girls with premature adrenarche: a risk factor for polycystic ovary syndrome. AB - Recently, the modest hyperandrogenism of premature adrenarche has been linked to the more severe hyperandrogenism of polycystic ovary syndrome (PCOS). The presence of hyperandrogenism, obesity, acanthosis nigricans, hyperinsulinism and reduced insulin sensitivity in girls with premature adrenarche and in women with PCOS suggests that the two conditions might have a common mechanism. As the early appearance of pubic hair might be the first presentation of hyperandrogenism and insulin resistance, these girls should have continued. PMID- 18406247 TI - The molecular and clinical spectrum of 3beta-hydroxysteroid dehydrogenase deficiency disorder. AB - Severe 3beta-hydroxysteroid dehydrogenase (3beta-HSD) deficiency in the adrenals and gonads is a well known cause of salt-wasting and non-salt-wasting forms of congenital adrenal hyperplasia (CAH), male pseudohermaphroditism and mild androgen excess symptoms in children and older females. A mild spectrum of Delta5 steroid abnormality in young children with premature pubarche and older females with hirsutism and menstrual disorder was presumed to be due to a mild variant of 3beta-HSD deficiency CAH. Recent studies of the type II 3beta-HSD gene encoding adrenal and gonadal 3beta-HSD have indicated that only the severe 3beta-HSD deficiency CAH results from a deleterious mutation in the gene. This indicates that the mild Delta5 steroid abnormality is not due to a variant of 3beta-HSD deficiency CAH. The hormonal criteria for bona fide mild variants of 3beta-HSD deficiency and etiology of mild Delta5 steroid abnormality in the patients remain to be investigated. PMID- 18406248 TI - Radioiodine-induced thyroid cancer: Studies in the aftermath of the accident at Chernobyl. AB - While a great deal is known about the relationship between external radiation exposure and thyroid cancer, much less is known about the oncogenic effects of internal radiation exposure from isotopes of iodine. The accident at the Chernobyl nuclear power plant released massive quantities of radioiodine isotopes into the atmosphere. The large number of ensuing thyroid cancers in exposed children leaves little doubt that these malignancies have occurred as a result of the accident. However, carefully planned epidemiological studies are needed to confirm that these are due predominantly to I-131 exposure, to determine the dose response relationship, to monitor for continuing effects and to evaluate other contributing factors. Preliminary evidence indicates that there is a distinct pattern of somatic genetic changes in the thyroid cancers from the Chernobyl area. PMID- 18406249 TI - Prolactin receptor gene diversity: structure and regulation. AB - The diverse functionality of prolactin and the wide expression of the prolactin receptor suggest a complex system regulated by this polypeptide hormone. Different hormone and receptor forms, as well as differential signal transduction pathways, contribute to the functional diversity of prolactin's actions. The heterogeneity of rat prolactin receptor gene transcripts in their 5'-untranslated region has led to the recognition of multiple and tissue-specific utilization of prolactin receptor gene promoters in gonadal and non-gonadal tissues. These findings have provided insights into the molecular bases for the diversity of prolactin's actions. It is now clear that cellular responsiveness to prolactin can be regulated through differential promoter control of the expression of the surface receptors for prolactin in different target tissues. PMID- 18406250 TI - Thyroid hormone receptor gene knockouts. AB - The thyroid hormone receptor genes, TRalpha and TRbeta, differ in developmental expression and tissue distribution. TRbeta knockout mice have goiter, elevated thyroid hormone and TSH levels, and a functional auditory defect. In contrast, mice with TRalpha 1/alpha2 inactivation have thyroid hypoplasia, low serum thyroid hormone levels, growth arrest and delayed small intestine maturation. Mice with selective TRalpha1 inactivation have apparent normal growth and development, but have bradycardia and reduced body temperature. The dramatic differences between these mice with TRbeta and TRalpha gene inactivations indicate the differential function of these genes. The influence of these gene inactivations on thyroid-stimulating hormone regulation is central to the resulting phenotypes. PMID- 18406251 TI - Calcitonin Gene-related Peptide (CGRP) is a Mediator of Vascular Adaptations During Hypertension in Pregnancy. AB - Hypertension during pregnancy is a serious obstetrical problem, and the pathophysiology of pre-eclampsia is unclear. Evidence is presented that calcitonin gene-related peptide (CGRP) plays a role in the vascular adaptations that occur during pregnancy. In addition, CGRP might be critical for compensatory vasodilatory changes that occur in response to hypertensive insults during pregnancy, and these compensatory mechanisms might require adequate progesterone activity. Thus, we suggest that an inadequate CGRP-mediated compensatory vasodilatory (adaptative) response may play a role in the pathophysiology of hypertensive disorders during pregnancy. PMID- 18406252 TI - Leptin: a molecule integrating somatic energy stores, energy expenditure and fertility. AB - The signaling of fat mass to central nervous system (CNS) regulators of food intake, energy expenditure and fertility has been inferred by experimental physiologists for over 75 years. The ability to modify such phenotypes based upon the status of body energy stores (fat) has critical survival value and, therefore, has been the object of potent selection pressure in evolution. The recent molecular cloning of the mouse ob mutation and the subsequent elucidation of the fundamentals of its regulatory physiology has identified a protein secreted by adipocytes, leptin, as a plausible candidate for a humoral signal with the requisite endocrinology and neurobiology. PMID- 18406253 TI - Xenoestrogens: the emerging story of bisphenol a. AB - Xenoestrogens are chemicals with diverse structure that mimic estrogen. Bisphenol A (BPA), a monomer of polycarbonate and epoxy resins, has been detected in canned food and human saliva. BPA stimulates cell proliferation and induces expression of estrogen-responsive genes in vitro, albeit with a relatively low potency. In vivo, BPA increases prolactin release and stimulates uterine, vaginal and mammary growth and differentiation. BPA shares similarities in structure, metabolism and action with diethylstilbestrol (DES), a known human teratogen and carcinogen. This review considers the hypothesis that BPA is converted in vivo to hydroxylated metabolite(s) with enhanced estrogenicity and genotoxicity. PMID- 18406254 TI - Androgen excess disorders in women. AB - Edited by Ricardo Azziz, John E. Nestler and Didier Dewailly. Philadelphia, Lippencott-Raven, 1997, $152.00/ pound116.00 (xxv +831 pages), ISBN 0-397-51721 1. PMID- 18406255 TI - Mineralocorticoid madness. AB - Adrenal Glands, Vascular System and Hypertension Edited by G.P. Vinson and D.C. Anderson. Bristol, UK, Society for Endocrinology, $95.00/ pound50.00 (312 pages), ISBN 1-898099-07-3. PMID- 18406256 TI - TSH Receptor Mutations and Thyroid Disease. AB - Mutations of the thyrotropin receptor (TSHr) can be loss of function or gain of function. Loss-of-function mutations can affect a variety of loci in the TSHr gene. Their most common manifestation is resistance to TSH; they may also be the cause of a subset of cases of congenital hypothyroidism. Gain-of-function mutations are of greater theoretical interest. Somatic mutations constitutively activating the TSHr are the major cause of benign toxic thyroid adenomas, and of some cases of multinodular goiters. They underlie hereditary toxic thyroid hyperplasia, and have been found in cases of sporadic congenital non-autoimmune hyperthyroidism. A role for TSHr polymorphisms in Graves' disease has not been documented. PMID- 18406257 TI - Molecular Properties of the CRF Receptor. AB - Research into the biology of corticotropin-releasing factor (CRF) has been intensified significantly by the structural characterization of the CRF receptor (CRF-R). Two receptor subtypes, CRF-R1 and CRF-R2, and three functional splice variants of CRF-R2 have been discovered. It appears that ligand binding requires interaction of the N-terminal domain with one or two other extracellular domains of the CRF-R. In contrast to the mammalian CRF-R1, the frog CRF-R1 discriminates between naturally occurring CRF-like peptides. PMID- 18406258 TI - Central nervous system effects of leptin. AB - Evidence exists demonstrating the importance of leptin in the control of energy homeostasis, feeding behavior and reproductive function. Leptin receptors are localized in several regions of the brain implicated in regulation of energy balance and reproductive function, including the arcuate nucleus/median eminence, paraventricular nucleus, and ventromedial nucleus. Administration of exogenous leptin has been shown to alter function of the hypothalamic-pituitary-adrenal axis and stimulate gonadotropin release through hypothalamic and pituitary actions. Results from in situ hybridization studies demonstrate the ability of leptin to modulate the expression of key neuropeptides (neuropeptide Y, corticotropin-releasing hormone) implicated in the regulation of energy homeostasis. This suggests that leptin is an important component in the neuroendocrine transmission line that regulates appetite, energy balance and reproduction. PMID- 18406259 TI - The androgen receptor as mediator of gene expression and signal transduction pathways. AB - The current model of action of androgens involves activation of a cytoplasmic receptor that migrates into the nucleus to regulate the expression of specific genes, either positively or negatively. While positive regulation requires direct binding of the receptor to DNA, negative regulation occurs mainly through protein protein interactions of the receptor and other transcription factors. More recent findings have shown that the receptor also mediates non-conventional responses attributed hitherto only to activated growth factor receptors. These actions proceed principally through activation of cytoplasmic kinases and they suggest that in addition to its genomic functions, the androgen receptor also regulates non-genomic processes. PMID- 18406260 TI - gp130-Related Cytokines and Their Receptors in the Pituitary. AB - gp130-Related cytokines such as interleukin-6 and leukemia-inhibitory factor (LIF) act on the adenohypophysis in a paracrine manner, affecting both its differentiation and the function of specific cell types, notably the proopiomelanocortin (POMC) cells. They act on POMC cells in synergism with corticotrophin-releasing hormone, inducing ACTH secretion. gp130-Related cytokines as well as their receptors are expressed in the pituitary. LIF knockout mice show reduced stress-induced ACTH secretion, which can be restored by LIF replacement, suggesting a physiologic role for LIF. PMID- 18406261 TI - The Role of DAX-1 in Reproduction. AB - Mutations in a gene referred to as Dax-1 cause an X-linked form of adrenal hypoplasia congenita (AHC). The disorder is limited to males and is characterized by neonatal adrenal insufficiency and failure to undergo puberty because of hypogonadotropic hypogonadism. Consistent with these clinical manifestations, the Dax-1 gene is expressed in the adrenal gland, gonads, hypothalamus and pituitary gland. The DAX-1 protein is structurally related to orphan nuclear receptors, although it lacks the characteristic zinc finger DNA-binding domain that is highly conserved in other members of this family. Dax-1 has been shown to repress the transcription of genes that are regulated by another nuclear receptor, steroidogenic factor-1 (SF-1). AHC mutations in Dax-1 eliminate its repressive activity. Genetic testing for Dax-1 mutations will enhance our ability to diagnose and treat AHC. Studies of the biological role of Dax-1 will provide new insights into the development and function of the adrenal gland and the reproductive axis. PMID- 18406262 TI - Regulatory Actions of Insulin-like Growth Factor-binding Proteins. AB - The six insulin-like growth factor-binding proteins (IGFBPs) are important regulators of insulin-like growth factor (IGF) action. Circulating high molecular weight complexes that contain IGF and IGFBP-3 restrict IGF bioavailability, and excess IGFBPs inhibit IGF action by forming biologically inactive complexes. IGFs can be released from these complexes by proteolysis. Potentiation of IGF activity might occur under specific circumstances, and involves the slow dissociation of IGFs from IGFBP complexes localized in the pericellular space, whose affinity has been reduced by dephosphorylation or association with the cell surface or extracellular matrix. Several IGFBPs or IGFBP fragments also have activities that do not involve IGFs or IGF receptors. The mechanisms by which IGFBPs regulate IGF action and exert their independent actions will be examined. PMID- 18406263 TI - Phenotype and genotype of osteoporosis. AB - A top priority in osteoporosis research is the identification of the structural basis and the genetic factors that contribute to variations in the risk for fragility fractures. This review summarizes current knowledge regarding the genetic determination of fragility fractures and bone mass measurements, and the complex phenotypes and genotypes of osteoporosis. PMID- 18406264 TI - Recent advances in the management of adrenal incidentalomas. AB - Modern imaging techniques have made adrenal incidentaloma a relatively common diagnostic problem. When an incidental adrenal mass is found, the clinician must try to determine its etiology and functionality, and the likelihood of malignancy. This task is complicated further in patients with a history of extra adrenal malignancy. In this article, we present a review of the literature and propose a diagnostic algorithm for management of adrenal incidentalomas. PMID- 18406265 TI - Insulin secretion, insulin sensitivity and diabetes in black children. AB - Historically, type 2 diabetes has been considered rare in the pediatric population. However, over the last decade, there has been a disturbing upswing in the rate of non-type 1 diabetes in the pediatric age group, particularly adolescents, with a greater proportion of Black children being affected. In this review, the following questions will be addressed: (1) what are the clinical characteristics of youth-onset atypical diabetes, (2) how common is it, (3) what are the risk factors, and (4) how should it be treated? PMID- 18406266 TI - Dyslipidaemia in Adult Growth Hormone (GH) Deficiency and the Effect of GH Replacement Therapy: A Review. AB - Adult growth hormone (GH) deficiency is associated with a lipid profile known to be related to atherosclerosis. GH replacement therapy improves the lipid profile with the exception of lipoprotein (a) concentrations, which tend to increase after GH therapy. Plasma lipid concentrations depend on its plasma carriers, the lipoproteins. Possible mechanisms involved in the dyslipidaemia of GH-deficient patients and the effects of GH replacement therapy are discussed with a special focus on hepatic lipoprotein metabolism. PMID- 18406267 TI - Menopause used to be considered normal. AB - Mishell's Textbook of Infertility, Contraception, and Reproductive Endocrinology. Fourth Edition Edited by Rogerio A. Lobo, Daniel R. Mishell, Jr, Richard J. Paulson and Donna Shoupe. Malden, MA, Blackwell Science, 1997, $175.00 (884 pages), ISBN 0-86542-385-7. PMID- 18406268 TI - The future on ice. AB - Reproductive Tissue Banking: Scientific Principles Edited by Armand M. Karow and John D. Critser. San Diego, Academic Press, 1997, $99.00 (xviii+472 pages), ISBN 0-12-399770-4. PMID- 18406269 TI - The functions of calcium across phyla. AB - The Comparative Endocrinology of Calcium Regulation Edited by C. Dacke, J. Danks, I. Caple and G. Flik. Bristol, UK, Society for Endocrinology, 1996, $85.00/ pound40.00 (220 pages), ISBN 1-898099-08-1. PMID- 18406271 TI - Guanylyl cyclases: approaching year thirty. AB - Since its discovery in 1963, cyclic GMP (cGMP) has been shown to be a ubiquitous second messenger. The enzymes that catalyze the formation of cGMP from GTP, guanylyl cyclases, exist in soluble and particulate isoforms. An explosion in the number of known isoforms, gene disruption, identification of new inhibitors and activators and finally the resolution of the structure of adenylyl cyclases have all provided important clues about the structure and function of guanylyl cyclases. This article gives a brief review of the recent developments in the field of guanylyl cyclase research. PMID- 18406272 TI - Familial forms broaden the horizons for primary aldosteronism. AB - The identification of familial forms of primary aldosteronism (PAL) has led to its detection in relatives of affected patients not suspected previously of having PAL. Many are normokalemic and some are even normotensive. This broadens the spectrum of PAL, permitting the study of its evolution and of intervention with specific therapy when hypertension develops. The genetic basis of one form involves steroid biosynthetic enzymes and the other form predisposes to hyperplasia and benign neoplasia. PMID- 18406273 TI - The broad spectrum of inherited growth hormone insensitivity syndrome. AB - Growth hormone (GH) insensitivity syndrome (GHIS) results in severe short stature and metabolic disturbances, but when this disorder is studied in more detail it is clear that there is marked clinical and biochemical heterogeneity. Many genetic defects of the GH receptor have been reported in inherited GHIS, but it now seems likely that some cases might be the result of defects of other genes or of links in the post-receptor cascade. PMID- 18406274 TI - Growth hormone treatment of short children born small for gestational age. AB - Short children born small-for-gestational-age (SGA) appear to be at an increased risk of having a poly-endocrinopathy, including a degree of growth hormone (GH) deficiency and/or insulin-like growth factor 1 (IGF-1) resistance. Among GH deficient children, those born SGA present a lower growth response to GH therapy than those not born SGA. The growth response of short SGA children to GH treatment does not appear to depend significantly on the secretory status of GH (as judged by provocative testing), indicating that the SGA condition (IGF-1 resistance) predominates over the availability of endogenous GH in determining the short stature of the majority of these children. When a higher than replacement dose of GH is administered, the growth response of short SGA children matches that of GH-deficient non-SGA children, indicating that the IGF-1 resistance towards growth can be overcome, and that a normal stature can be obtained, at least throughout childhood. It is anticipated that, increasingly, the indications and the doses for GH therapy in children will become interlinked with the emerging principles of endocrine programming in early life. PMID- 18406275 TI - Recent Advances in the Study of hsp90 Structure and Mechanism of Action. AB - The 90kDa heat shock protein, hsp90, is a major molecular chaperone of the cell that appears to have particular significance to cellular regulatory processes. New tools and approaches have revealed a number of target proteins for hsp90, most of which are protein kinases or transcription factors. While the mechanism of action of hsp90 is not well understood, reasonable models have emerged describing some functional domains of this protein, the importance of conformational transitions for its activity and its role within a multi-component chaperoning pathway of the cell. PMID- 18406276 TI - Studies with Purified Chaperones Advance the Understanding of the Mechanism of Glucocorticoid Receptor-hsp90 Heterocomplex Assembly. AB - The study of the 9S, untransformed state of steroid receptors has led to the discovery of a multiprotein chaperone system that assembles heterocomplexes between hsp90 and a variety of proteins involved in signal transduction. Using the formation of glucocorticoid receptor (GR)-hsp90 heterocomplexes as a model, we have reconstituted a fully functional heterocomplex assembly system from purified components. The basic assembly system requires four proteins-hsp90, hsp70, p60/Hop and hsp40-to assemble GR-hsp90 heterocomplexes, which are then stabilized by the hsp90-interacting protein p23. The four proteins can self assemble into an hsp90-p60/Hop-hsp70-hsp40 complex that we call a foldosome. Foldosomes isolated from reticulocyte lysate or formed from purified proteins open up a steroid-binding pocket to create a high-affinity steroid-binding state of the GR. We describe here the systematic reconstitution of the hsp90-based chaperone machinery and develop a model of the receptor-hsp90 heterocomplex assembly mechanism. PMID- 18406278 TI - The fountain of youth? AB - DHEA and Sex Steroids. Intracrinology in Target Tissues Edited by F. Labrie, A. Belanger, J. Simard, V. Luu-The and C. Labrie. Bristol, UK, Society for Endocrinology, 1996, $69.95/ pound30.00 (265 pages), ISBN 1-898099-3. PMID- 18406279 TI - A ;C' for the Pancreatic ABCs. AB - Clinical Pathology of Pancreatic Disorders Edited by John A. Lott. Totowa, Humana, 1997, $99.50 (ix+218 pages), ISBN 0-896-03475-5. PMID- 18406280 TI - A search for gems in a sea of shifting sand. AB - Localized Prostate Cancer. Advances in Diagnosis and Treatment Edited by F. Labrie, F. Lee and D. Crawford. Bristol, UK, Society for Endocrinology, 1996, $59.95/ pound25.00 (148 pages), ISBN 1-898099-11-1. PMID- 18406281 TI - The vitamin d receptor gene and calcium metabolism. AB - Dietary Ca(2+) is essential for the development and maintenance of bone mineral mass. The vitamin D endocrine system plays a fundamental role in the regulation of Ca(2+) homeostasis, and mutations affecting genes implicated in vitamin D metabolism or vitamin D receptor (VDR) functions are responsible for severe alterations in skeletal growth. In addition, vitamin D is also implicated in the pathophysiology and treatment of adult bone disorders associated with impaired mineralization of bone matrix. Very recently, common polymorphisms in the 3'- and 5'-end region of the VDR gene have been suggested as possible determinants of bone mineral mass and, hence, of the risk of osteoporosis. None of these polymorphisms appear to be associated unequivocally with bone mineral mass or biochemical variables of Ca(2+) and phosphate metabolism, except perhaps VDR 3' end polymorphisms before puberty. As these associations are so inconsistent, interactions with environmental factors, particularly dietary intake, and with other polymorphisms have to be considered. PMID- 18406282 TI - 17beta-hydroxysteroid dehydrogenases: physiological roles in health and disease. AB - Androgens and estrogens play crucial roles in the growth and development of sex organs. Interconversion of these hormones between biologically active and inactive forms is catalyzed by 17beta-hydroxysteroid dehydrogenase (17beta-HSD) isozymes. Aberrations in the regulation or expression of the various 17beta-HSD isoforms has been implicated in the genesis/progression of hormonally dependent cancers of various tissues, including ovary, breast and prostate; in the predisposition of women with upper body obesity to several types of diseases, such as non-insulin dependent diabetes mellitus; and in the abnormal development of sexually ambiguous individuals, as seen in 17beta-HSD-deficient male pseudohermaphrodites. Of the five known 17beta-HSD isozymes, deleterious mutations in the type 3 isoform were found to give rise to male pseudohermaphroditism. The 16 mutations characterized to date include 12 missense mutations, three splice junction mutations, and one small deletion that results in a downstream premature stop codon. 17beta-HSD has also been studied in other species. The most notable species difference observed is the placental expression in humans of the 17beta-HSD type 1 isoform. PMID- 18406283 TI - Endocrine, paracrine and cellular regulation of postnatal anti-mullerian hormone secretion by sertoli cells. AB - Anti-Mullerian hormone (AMH) is produced by Sertoli cells from early foetal life to puberty, when it is downregulated by the synergistic action of intratesticular testosterone and meiotic germ cells. During foetal and neonatal life, testosterone is incapable of inhibiting AMH, owing to the lack of androgen receptor expression in Sertoli cells. In the absence of an androgen inhibitory effect, follicle-stimulating hormone increases testicular AMH output. PMID- 18406284 TI - Lessons from the genetics of laron syndrome. AB - In the decade since the cloning and sequencing of the growth hormone receptor (GHR) and the recognition that the circulating GH-binding protein (GHBP) is structurally identical to the extracellular domain of the GHR, 34 mutations have been described. These include one deletion, eight nonsense mutations, eleven missense mutations, four frameshift mutations and ten splice mutations. More than half of the 131 patients with Laron syndrome whose molecular defects have been identified comprise the Ecuadorian cohort who share a single splice mutation. Variable expression of different homozygous or compound heterozygous defects of the GHR is no greater than the variation within a genetically homogeneous population. Some features, such as birth size and intelligence, are unlikely to be affected by GHRD. Greater understanding of the genetics, physiology, and clinical expression of abnormalities in the GH-GHR-IGF-I (insulin-like growth factor I) axis necessitates a reconsideration of the classification of GH insensitivity (GHI). PMID- 18406285 TI - Prenatal treatment of congenital adrenal hyperplasia. AB - Prenatal diagnosis of congenital adrenal hyperplasia (CAH) is accurate and prenatal therapy is effective in significantly reducing or even eliminating virilization of females affected by CAH, sparing these children the consequences of genital surgery, sex missassignment and gender confusion. However, both the physical and psychological development of these children and the possibility of long-range adverse effects in the mothers need to be evaluated further. Prospective multicentre studies covering several decades are being designed. PMID- 18406286 TI - Prenatal treatment of congenital adrenal hyperplasia: a promising experimental therapy of unproven safety. AB - The long-term safety of the prenatal treatment of congenital adrenal hyperplasia (CAH) has not been established, and it remains an experimental therapy that raises unique ethical questions. Prenatal treatment should be performed only according to controlled, prospective, peer-reviewed protocols carried out according to strict scientific and ethical standards. Such studies must incorporate detailed, decades-long follow-up to permit accurate appraisal of its efficacy and safety, especially among the seven of eight fetuses who receive unneeded treatment. PMID- 18406287 TI - Prenatal treatment of congenital adrenal hyperplasia: a commentary. PMID- 18406288 TI - Protein kinase C and exocytosis in Mammalian spermatozoa. PMID- 18406290 TI - Species differences in hormones: yes, it does matter. AB - Laboratory Animal Endocrinology. Hormonal Action, Control Mechanisms and Interactions with Drugs By David D. Woodman. Chichester, UK, Wiley, 1997, $189.95 (vii+567 pages), ISBN 0-471-97262-2. PMID- 18406289 TI - Protein kinase C and exocytosis in Mammalian spermatozoa. PMID- 18406291 TI - How to build a pancreas. AB - Pancreatic Growth and Regeneration Edited by Nora Sarvetnick. Basel, Karger, 1997, CHF113.0/DEM135.00/$98.00 (xiv+250 pages) ISBN 3-8055-6618-2. PMID- 18406292 TI - Small brain neuropeptides. AB - Recent Advances in Arthropod Endocrinology Edited by Geoffrey M. Coast and Simon G. Webster. Cambridge, Cambridge University Press, 1998, $110.00 (xviii+406 pages), ISBN 0-521-59113-9. PMID- 18406294 TI - Current concepts of the mechanisms of menstruation: a normal process of tissue destruction. AB - Current data support the contention that menstruation is the consequence of the actions of matrix metalloproteinases (MMPs) in the endometrium, and that these enzymes are stimulated by the products of an inflammatory process. A number of MMPs, capable of degrading both interstitial matrix and basement membrane components, have been localized to perimenstrual endometrium, and the focal nature of their production suggests local regulation. Emphasis is placed on important relationships between cells of the immune system (specifically, mast cells, eosinophils, neutrophils and macrophages), and the local production and activation of MMPs, whose degradative actions lead to loss of integrity of blood vessels, destruction of endometrial interstitial matrix and the resultant bleeding characteristic of menstruation. PMID- 18406295 TI - Prostate-specific Antigen: Its Usefulness in Clinical Medicine. AB - Prostate-specific antigen (PSA) was discovered about 20 years ago and over the last decade it has become the premier tumor marker for diagnosis, monitoring and prognosis of prostatic carcinoma. PSA is now considered to be the best tumor marker available in clinical medicine. It is the only tumor marker that has been approved by the Food and Drug Administration of the USA for mass screening: for the purpose of diagnosing early prostatic carcinoma. Among the newest developments in the field are the discovery of the molecular forms of PSA in serum, the development of ultrasensitive assays that allow better monitoring of patients after radical prostatectomy, and the discovery of non-prostatic PSA. Indeed, there are indications that PSA might be useful for the diagnosis and prognosis of breast cancer. The genomic structure of PSA and other human kallikrein genes and the regulation of their expression has recently been elucidated. Currently, the PSA promoter and enhancer are being investigated in connection with gene therapy in prostatic tissue. Efforts are now underway to supplement the clinical value of PSA measurements with additional prostatic markers, including human kallikrein 2 (hK2) and prostate-specific membrane antigen (PSMA). PMID- 18406296 TI - Molecular mechanism of androgen action. AB - Androgens affect the growth and development of a wide variety of cell types in both males and females and produce their effects by binding to androgen receptors, which modulate the transcription of specific genes. Testosterone is the major active androgen circulating in blood, but in many tissues it is metabolized by 5alpha-reductase to 5alpha-dihydrotestosterone, which binds to and activates the androgen receptor. Androgen receptors are members of the nuclear receptor family of transcription factors, and these nuclear receptors control transcription by recruitment of a variety of co-activators and co-repressors. Mutations in the androgen receptor and 5alpha-reductase can affect male sexual development. 5alpha-Reductase is also critical for parturition and fetal survival in mice. Inhibitors of 5alpha-reductase are being used increasingly to treat some androgen-dependent disorders. Because androgens also suppress the growth of certain cancer cells, they might also have a role in treating prostate cancer. PMID- 18406297 TI - The Role of Phosphatidylinositol Transfer Proteins (PITPs) in Intracellular Signalling. AB - Phosphatidylinositol transfer protein (PITP) has been identified as a key player in numerous signalling pathways relying on phosphatidylinositol (PI) metabolites. Although its cellular function is most likely linked to its PI/phosphatidylcholine (PC) transfer activity-an in vitro activity shared by all known PITPs-this feature cannot explain all findings from studies with PITP. Here, we review evidence suggesting that one of the main functions of PITP in cellular signalling is to present PI to lipid kinases for localized production of phosphatidylinositol (4,5)-bisphosphate (PIP(2)), either to be used as a signalling molecule (for example, in exocytosis) or as a substrate (for example, by phospholipases). PMID- 18406298 TI - Corticotropin releasing hormone, receptor regulation and the stress response. AB - Corticotropin releasing hormone (CRH) coordinates behavioral, autonomic and hormonal responses to stress, including activation of the hypothalamic-pituitary adrenal (HPA) axis with stimulation of adrenocorticotropin (ACTH) and glucocorticoids. Differential changes of expression of CRH and vasopressin(VP) in the parvicellular hypothalamic paraventricular nucleus (PVN), as well as regulation of CRH and VP receptors, are critical for the responsiveness of the HPA axis during stress. Pituitary CRH receptor (CRH-R)expression and content is controlled by the coordinated action of CRH, VP and glucocorticoids. Marked changes in hypothalamic and pituitary CRH-R expression support a key regulatory role for CRH in the HPA axis and the integrated stress response. PMID- 18406299 TI - Lispro insulin: benefits and limitations. AB - Lispro insulin is the first synthetic insulin analog commercially available for the treatment of diabetes. Its rapid absorption and short half-life results in both advantages and disadvantages to the patient with diabetes. Understanding the physiology of normal insulin secretion and the pathophysiology of diabetes is paramount to optimizing Lispro insulin in patients with diabetes. PMID- 18406300 TI - Variations on a theme in g. AB - G Proteins, Receptors, and Disease Edited by Allen M. Spiegel. Totowa, Humana, 1998, $135.00 (x+324 pages), ISBN 0-896-034305. PMID- 18406301 TI - Exploring protein interactions. AB - Protein Targeting Protocols Edited by Robert A. Clegg. Totowa, Humana, 1998, $79.50 (xii+ 331 pages), ISBN 0-896-03450-X. PMID- 18406302 TI - Hormone signaling pathways: planes, trains, and automobiles... AB - Hormones and Signaling, Volume 1 Edited by Bert O'Malley. San Diego, Academic Press, 1998, $69.95 (xv+364 pages), ISBN 0-23-312411-5. PMID- 18406304 TI - The brain as an endocrine target for Peptide hormones. AB - Unlike circulating steroid hormones, which have a relatively unhindered passage into the central nervous system, blood-borne peptides are usually restricted by the blood-brain barrier. Some circulating peptides, such as angiotensin II, atrial natriuretic peptide and relaxin, influence central neural pathways subserving cardiovascular and body fluid homeostasis by acting on neurons in the subfornical organ, organum vasculosum of the lamina terminalis and area postrema, all of which lack a blood-brain barrier. There are some circulating peptides such as insulin and leptin that are transported from the bloodstream across cerebral blood vessel walls into sites in the hypothalamus that have appropriate neural connections to influence food intake and sympathetic control of brown fat. PMID- 18406305 TI - Ca(2+) receptor from brain to gut: common stimulus, diverse actions. AB - An extracellular Ca(2+)-sensing receptor (CaR) plays central roles in Ca(2+) homeostasis by regulating parathyroid hormone (PTH)secretion and renal Ca(2+) handling. The CaR is also expressed in intestine and bone, where its functions in mineral metabolism are not yet well defined. The receptor is also present in various types of cells seemingly uninvolved in systemic mineral ion homeostasis (such as neuronal and glial cells in the brain and various epithelial cells), where its actions are poorly understood but might involve the regulation of local ionic homeostasis and/or diverse cellular processes, such as cellular differentiation and proliferation. PMID- 18406306 TI - Cancer in acromegaly. AB - In recent years, it has become increasingly recognized that acromegaly predisposes to a variety of cancers, particularly colorectal and breast cancers, and perhaps haematological malignancies. However, these associations have been based mostly on small epidemiological surveys, and a propensity towards other malignancies might also become apparent in the future. This review assesses these three malignancies together with those of the thyroid and prostate, and discusses their pathogenesis, concentrating on the role of the growth hormone/insulin-like growth factor I (IGF-I) axis. PMID- 18406307 TI - Leukemia in children treated with growth hormone. AB - An association between treatment with growth hormone (GH) and the development of leukemia was described in 1988. This perceived association is best explained by the fact that there are more children with GH deficiency (GHD) with risk factors predisposing them to leukemia than in the general population. These factors include previous cancers and their treatment, as well as co-existing conditions such as Down, Bloom and Fanconi syndromes. Examination of large databases of GH treated individuals shows that GH-treated patients without these risk factors do not have an increased incidence of leukemia. PMID- 18406308 TI - The roles of oestrogen in the male. AB - Roles for oestrogens in brain masculinization/sexual behaviour, regulation of follicle-stimulating hormone (FSH)secretion and Leydig cell development and function are well established. However, the widespread distribution of oestrogen receptors alpha and beta in reproductive and other tissues of the male, and findings from human males or transgenic animals in which the genes coding for these receptors or for aromatase are non-functional, are changing our perception of the roles of oestrogen in the male. Aspects of pubertal development in boys (growth of the long bones, their mineralization and epiphyseal closure) attributed to the actions of androgens are now recognized as being mediated in part by oestrogens. Oestrogens also play a role (probably vasodilatatory) in the cardiovascular system of the male. Within the reproductive system, oestrogens have been shown to play a role in the regulation of fluid resorption from the efferent ducts and appear to be important in the structural and functional development of the Wolffian/excurrent duct system, as well as that of the prostate; inappropriately low or high oestrogen exposure during development can cause permanent changes to these tissues, which may lead to disorders of spermatogenesis and infertility. Sertoli cells and certain germ cells in the testis are also targets for oestrogen action. Many other tissues (adipose, kidney, thymus/immune system, skin, gut and muscle) are oestrogen targets in the male. Based on these findings and the widespread distribution of aromatase, it is argued that many of the effects of oestrogens in the male might stem from its local production and action and, furthermore, that the balance in action between androgens and oestrogens might be of central importance at many oestrogen target sites. PMID- 18406310 TI - Monitoring phospholipid signaling pathways: recipes from the experts. AB - Phospholipid Signaling Protocols Edited by Ian M. Bird. Totowa, Humana, 1998, $79.50 (xii +380 pages), ISBN 0-896-03491-7. PMID- 18406309 TI - Endothelins as autocrine regulators of tumor cell growth. AB - Endothelin 1 (ET-1) is produced by several types of human cancer cells and has been proposed to participate in tumor development or progression by exerting autocrine or paracrine actions on neoplastic cells and their surrounding stromal cells. Recently, an ET-1-mediated autocrine loop has been implicated in the growth of ovarian tumor cells. The co-expression of ET-1 and ET(A) receptors, with consequent activation of growth signaling pathways in human ovarian carcinoma cells, constitutes a mechanism for the autocrine regulation of tumor cell growth. Such findings also provide a basis for further investigation of the role of tyrosine phosphorylation in ET-1-regulated growth responses in ovarian tumor cells. The overexpression of ET-1 and its receptor in cancer cells may serve as a tumor marker, and could provide potential targets for therapy. PMID- 18406312 TI - ERbeta: a novel estrogen receptor offers the potential for new drug development. AB - There is increasing interest in developing better drugs for improving the health of women. Because of the multiple target organs for estrogens and the occurrence of both beneficial and unwanted effects during treatment, the key to improvement in drug therapy is the development of estrogen receptor modulators with better tissue selectivity. The recent discovery that there are not one but two estrogen receptors, ERalpha and ERbeta, each with its unique tissue distribution and with differing and sometimes opposing actions on certain genes, promises new hope for the development of novel, tissue-selective estrogens. Our present knowledge of the tissue distribution of ERalpha and ERbeta suggests that development of selective therapies for treatment and/or prevention of menopausal symptoms, osteoporosis, cardiovascular disease, type II diabetes, Alzheimer's disease and urinary incontinence is an achievable goal in the foreseeable future. Furthermore, it is possible that future estrogen therapy might be beneficial for men. PMID- 18406313 TI - Prohormones: their clinical relevance. AB - The majority of peptide hormones and neuropeptides are synthesized as precursors, which are cleaved in a sequence-specific and tissue-specific manner to yield the biologically active peptides. There has been considerable progress in the past ten years in understanding the nature and mechanism of action of the prohormone convertases that cleave these prohormones. Evidence from knockout technology and clinical examples of gene mutations has provided functional information on disruption of prohormone cleavage and the bioactivity of prohormones. There are specific examples of the clinical relevance of circulating prohormones, such as adrenocorticotrophin (ACTH) precursors and proinsulin. The central issues that still remain are: (1) What is the relative importance of each of the different processing pathways and processing enzymes in regulating hormone action? (2) How do the serum concentrations of prohormones compare with the mature hormone levels? (3) What are the biological consequences of prohormones in the circulation? PMID- 18406314 TI - The systematic screening and management of hypothyroidism and hyperthyroidism during pregnancy. AB - Altogether, thyroid function abnormalities during pregnancy can affect up to 10% of all women. The high prevalence of both hypo- and hyperthyroidism, the obstetrical repercussions associated with thyroid dysfunction in the mothers, as well as the potential role of maternal thyroid dysfunction as an influence on fetal development constitute solid arguments for a further increase of our knowledge of the pathophysiological processes underlying the alterations of thyroid function related to the pregnant state. In this review, the focus will be on the most clinically relevant aspects associated with hypothyroidism [autoimmune thyroid disorders (AITDs), subfertility, risk of miscarriage, risk of hypothyroidism in women with AITD and treatment of hypothyroid women] and with hyperthyroidism (clinical presentations during pregnancy, Graves' disease and its management, fetal hyperthyroidism in women with antithyroid-stimulating hormone receptor antibodies and gestational transient thyrotoxicosis associated with human chorionic gonadotropin stimulation of the maternal thyroid gland). I also propose a global strategy for the systematic screening of hypo- and hyperthyroidism in the pregnant state. PMID- 18406315 TI - The acute regulation of mineralocorticoid biosynthesis: scenarios for the StAR system. AB - The zona glomerulosa cell of the adrenal cortex produces mineralocorticoids in response to physiological stimuli (angiotensin II and extracellular K(+)) activating the Ca(2+) messenger system. The mechanisms underlying the generation of the Ca(2+) signal have been analyzed extensively and recent developments have contributed to bridging the gap between intracellular signals and activation of the biological function. This article summarizes the current knowledge on the intracellular targets of the Ca(2+) messenger, obtained mainly in bovine glomerulosa cells. Ca(2+) appears to exert a dual effect, both at the intramitochondrial level and at the nuclear level, where it activates steroidogenic acute regulatory protein (StAR) gene transcription. PMID- 18406316 TI - Nongenomic effects of 1alpha,25-dihydroxyvitamin D(3). AB - The hormonally active form of vitamin D, 1alpha,25-dihydroxyvitamin D(3), is the key molecule of the vitamin D endocrine system, which produces biological effects in about 30 target cell systems. Growing experimental evidence supports the hypothesis that these biological effects can be generated both by a signal transduction mechanism involving a nuclear receptor (nVDR) that modulates gene transcription, and via a nongenomic receptor located in the plasma membrane (mVDR), which modulates a complex signaling system involving the rapid opening of Ca(2+) channels. Some data reviewed herein also indicate that crosstalk between genomic and nongenomic pathways operates in several cell types, and suggest that the physiological role of the rapid, nongenomic actions might involve the regulation of hormone-mediated gene activation. PMID- 18406319 TI - Gene regulation: hacking the network on a sugar high. AB - In a recent issue of Molecular Cell, Kaplan et al. (2008) determine the input functions for 19 E. coli sugar-utilization genes by using a two-dimensional high throughput approach. The resulting input-function map reveals that gene network regulation follows non-Boolean, and often nonmonotonic, logic. PMID- 18406320 TI - Not just because it is there: aminoacyl-tRNA synthetases gain control of the cell. AB - In a recent issue of Molecular Cell, Jia et al. (2008) demonstrate that time controlled repression of interferon-induced transcripts depends upon the interaction between an RNA structure in these transcripts and protein domains harbored by a mammalian aminoacyl-tRNA synthetase. PMID- 18406321 TI - From DNA end chemistry to cell-cycle response: the importance of structure, even when it's broken. AB - In this issue of Molecular Cell, Barlow et al. (2008) show that not all DNA double-strand breaks are processed equally and that the chemical nature of DNA ends guides different paths to DNA repair. PMID- 18406322 TI - Poised polymerases: on your mark...get set...go! AB - Recent global analyses have determined that many Drosophila and human genes have engaged polymerase molecules trapped immediately downstream of promoters. These results strongly implicate RNA polymerase II elongation control as a major regulator of differentiation and development. PMID- 18406323 TI - Quantitative analysis of pathways controlling extrinsic apoptosis in single cells. AB - Apoptosis in response to TRAIL or TNF requires the activation of initiator caspases, which then activate the effector caspases that dismantle cells and cause death. However, little is known about the dynamics and regulatory logic linking initiators and effectors. Using a combination of live-cell reporters, flow cytometry, and immunoblotting, we find that initiator caspases are active during the long and variable delay that precedes mitochondrial outer membrane permeabilization (MOMP) and effector caspase activation. When combined with a mathematical model of core apoptosis pathways, experimental perturbation of regulatory links between initiator and effector caspases reveals that XIAP and proteasome-dependent degradation of effector caspases are important in restraining activity during the pre-MOMP delay. We identify conditions in which restraint is impaired, creating a physiologically indeterminate state of partial cell death with the potential to generate genomic instability. Together, these findings provide a quantitative picture of caspase regulatory networks and their failure modes. PMID- 18406324 TI - Translational regulation via L11: molecular switches on the ribosome turned on and off by thiostrepton and micrococcin. AB - The thiopeptide class of antibiotics targets the GTPase-associated center (GAC) of the ribosome to inhibit translation factor function. Using X-ray crystallography, we have determined the binding sites of thiostrepton (Thio), nosiheptide (Nosi), and micrococcin (Micro), on the Deinococcus radiodurans large ribosomal subunit. The thiopeptides, by binding within a cleft located between the ribosomal protein L11 and helices 43 and 44 of the 23S rRNA, overlap with the position of domain V of EF-G, thus explaining how this class of drugs perturbs translation factor binding to the ribosome. The presence of Micro leads to additional density for the C-terminal domain (CTD) of L7, adjacent to and interacting with L11. The results suggest that L11 acts as a molecular switch to control L7 binding and plays a pivotal role in positioning one L7-CTD monomer on the G' subdomain of EF-G to regulate EF-G turnover during protein synthesis. PMID- 18406325 TI - Unique contacts direct high-priority recognition of the tetrameric Mu transposase DNA complex by the AAA+ unfoldase ClpX. AB - Clp/Hsp100 ATPases remodel and disassemble multiprotein complexes, yet little is known about how they preferentially recognize these complexes rather than their constituent subunits. We explore how substrate multimerization modulates recognition by the ClpX unfoldase using a natural substrate, MuA transposase. MuA is initially monomeric but forms a stable tetramer when bound to transposon DNA. Destabilizing this tetramer by ClpX promotes an essential transition in the phage Mu recombination pathway. We show that ClpX interacts more tightly with tetrameric than with monomeric MuA. Residues exposed only in the MuA tetramer are important for enhanced recognition--which requires the N domain of ClpX--as well as for a high maximal disassembly rate. We conclude that an extended set of potential enzyme contacts are exposed upon assembly of the tetramer and function as internal guides to recruit ClpX, thereby ensuring that the tetrameric complex is a high-priority substrate. PMID- 18406326 TI - The double bromodomain proteins Brd2 and Brd3 couple histone acetylation to transcription. AB - Posttranslational histone modifications are crucial for the modulation of chromatin structure and regulation of transcription. Bromodomains present in many chromatin-associated proteins recognize acetylated lysines in the unstructured N terminal regions of histones. Here, we report that the double bromodomain proteins Brd2 and Brd3 associate preferentially in vivo with hyperacetylated chromatin along the entire lengths of transcribed genes. Brd2- and Brd3 associated chromatin is significantly enriched in H4K5, H4K12, and H3K14 acetylation and contains relatively little dimethylated H3K9. Both Brd2 and Brd3 allowed RNA polymerase II to transcribe through nucleosomes in a defined transcription system. Such activity depended on specific histone H4 modifications known to be recognized by the Brd proteins. We also demonstrate that Brd2 has intrinsic histone chaperone activity and is required for transcription of the cyclin D1 gene in vivo. These data identify proteins that render nucleosomes marked by acetylation permissive to the passage of elongating RNA polymerase II. PMID- 18406327 TI - Deletion of histone deacetylase 3 reveals critical roles in S phase progression and DNA damage control. AB - Histone deacetylases (HDACs) are enzymes that modify key residues in histones to regulate chromatin architecture, and they play a vital role in cell survival, cell-cycle progression, and tumorigenesis. To understand the function of Hdac3, a critical component of the N-CoR/SMRT repression complex, a conditional allele of Hdac3 was engineered. Cre-recombinase-mediated inactivation of Hdac3 led to a delay in cell-cycle progression, cell-cycle-dependent DNA damage, and apoptosis in mouse embryonic fibroblasts (MEFs). While no overt defects in mitosis were observed in Hdac3-/- MEFs, including normal H3Ser10 phosphorylation, DNA damage was observed in Hdac3-/- interphase cells, which appears to be associated with defective DNA double-strand break repair. Moreover, we noted that Hdac3-/- MEFs were protected from DNA damage when quiescent, which may provide a mechanistic basis for the action of HDAC inhibitors on cycling tumor cells. PMID- 18406328 TI - Differential regulation of the cellular response to DNA double-strand breaks in G1. AB - Double-strand breaks (DSBs) are potentially lethal DNA lesions that can be repaired by either homologous recombination (HR) or nonhomologous end-joining (NHEJ). We show that DSBs induced by ionizing radiation (IR) are efficiently processed for HR and bound by Rfa1 during G1, while endonuclease-induced breaks are recognized by Rfa1 only after the cell enters S phase. This difference is dependent on the DNA end-binding Yku70/Yku80 complex. Cell-cycle regulation is also observed in the DNA damage checkpoint response. Specifically, the 9-1-1 complex is required in G1 cells to recruit the Ddc2 checkpoint protein to damaged DNA, while, upon entry into S phase, the cyclin-dependent kinase Cdc28 and the 9 1-1 complex both serve to recruit Ddc2 to foci. Together, these results demonstrate that the DNA repair machinery distinguishes between different types of damage in G1, which translates into different modes of checkpoint activation in G1 and S/G2 cells. PMID- 18406329 TI - FACT-mediated exchange of histone variant H2AX regulated by phosphorylation of H2AX and ADP-ribosylation of Spt16. AB - The phosphorylation of histone variant H2AX at DNA double-strand breaks is believed to be critical for recognition and repair of DNA damage. However, little is known about the molecular mechanism regulating the exchange of variant H2AX with conventional H2A in the context of the nucleosome. Here, we isolate the H2AX associated factors, which include FACT (Spt16/SSRP1), DNA-PK, and PARP1 from a human cell line. Our analyses demonstrate that the H2AX-associated factors efficiently promote both integration and dissociation of H2AX and this exchange reaction is mainly catalyzed by FACT among the purified factors. The phosphorylation of H2AX by DNA-PK facilitates the exchange of nucleosomal H2AX by inducing conformational changes of the nucleosome. In contrast, poly-ADP ribosylation of Spt16 by PARP1 significantly inhibits FACT activities for H2AX exchange. Thus, these data establish FACT as the major regulator involved in H2AX exchange process that is modulated by H2AX phosphorylation and Spt16 ADP ribosylation. PMID- 18406330 TI - Retrotransposon Tf1 is targeted to Pol II promoters by transcription activators. AB - The LTR-retrotransposon Tf1 preserves the coding capacity of its host Schizosaccharomyces pombe by integrating upstream of open reading frames (ORFs). To determine which features of the target sites were recognized by the transposon, we introduced plasmids containing candidate insertion sites into S. pombe and mapped the positions of integration. We found that Tf1 was targeted specifically to the promoters of Pol II-transcribed genes. A detailed analysis of integration in plasmids that contained either ade6 or fbp1 revealed insertions occurred in the promoters at positions where transcription factors bound. Further experiments revealed that the activator Atf1p and its binding site were required for directing integration to the promoter of fbp1. An interaction between Tf1 integrase and Atf1p was observed, indicating that integration at fbp1 was mediated by the activator bound to its promoter. Surprisingly, we found Tf1 contained sequences that activated transcription, and these substituted for elements of the ade6 promoter disrupted by integration. PMID- 18406331 TI - Glycolytic enzyme GAPDH promotes peroxide stress signaling through multistep phosphorelay to a MAPK cascade. AB - Phosphorelay signaling of environmental stimuli by two-component systems is prevailing in bacteria and also utilized by fungi and plants. In the fission yeast Schizosaccharomyces pombe, peroxide stress signals are transmitted from the Mak2/3 sensor kinases to the Mpr1 histidine-containing phosphotransfer (HPt) protein and finally to the Mcs4 response regulator, which activates a MAP kinase cascade. Here we show that, unexpectedly, the glycolytic enzyme glyceraldehyde-3 phosphate dehydrogenase (GAPDH) physically associates with the Mcs4 response regulator and stress-responsive MAP kinase kinase kinases (MAPKKKs). In response to H2O2 stress, Cys-152 of the Tdh1 GAPDH is transiently oxidized, which enhances the association of Tdh1 with Mcs4. Furthermore, Tdh1 is essential for the interaction between the Mpr1 HPt protein and the Mcs4 response regulator and thus for phosphorelay signaling. These results demonstrate that the glycolytic enzyme GAPDH plays an essential role in the phosphorelay signaling, where its redox sensitive cysteine residue may provide additional input signals. PMID- 18406332 TI - Assembly and channel opening in a bacterial drug efflux machine. AB - Drugs and certain proteins are transported across the membranes of Gram-negative bacteria by energy-activated pumps. The outer membrane component of these pumps is a channel that opens from a sealed resting state during the transport process. We describe two crystal structures of the Escherichia coli outer membrane protein TolC in its partially open state. Opening is accompanied by the exposure of three shallow intraprotomer grooves in the TolC trimer, where our mutagenesis data identify a contact point with the periplasmic component of a drug efflux pump, AcrA. We suggest that the assembly of multidrug efflux pumps is accompanied by induced fit of TolC driven mainly by accommodation of the periplasmic component. PMID- 18406333 TI - Fluorescence intensity decays of 2-aminopurine solutions: lifetime distribution approach. AB - The fluorescent adenine analog 2-aminopurine (2AP) has been used extensively to monitor conformational changes and macromolecular binding events involving nucleic acids because its fluorescence properties are highly sensitive to changes in chemical environment. Furthermore, site-specific incorporation of 2AP permits local DNA and RNA conformational events to be discriminated from the global structural changes monitored by UV-Vis spectroscopy and circular dichroism. However, although the steady-state fluorescence properties of 2AP have been well defined in diverse settings, interpretation of 2AP fluorescence lifetime parameters has been hampered by the heterogeneous nature of multiexponential 2AP intensity decays observed across populations of microenvironments. To resolve this problem, we tested the utility of multiexponential versus continuous Lorentzian lifetime distribution models to describe fluorescence intensity decays from 2AP in diverse chemical backgrounds and within the context of RNA. Heterogeneity was introduced into 2AP intensity decays by mixing solvents of differing polarities or by adding quenchers under high viscosity to evaluate the transient effect. Heterogeneity of 2AP fluorescence within the context of a synthetic RNA hairpin was introduced by structural remodeling using a magnesium salt. In each case except folded RNA (which required a bimodal distribution), 2AP lifetime properties were well described by single Lorentzian distribution functions, abrogating the need to introduce additional discrete lifetime subpopulations. Rather, heterogeneity in fluorescence decay processes was accommodated by the breadth of each distribution. This approach also permitted solvent relaxation effects on 2AP emission to be assessed by comparing lifetime distributions at multiple wavelengths. Together, these studies provide a new perspective for the interpretation of 2AP fluorescence lifetime properties that will further the utility of this fluorophore in analyses of the complex and heterogeneous structural microenvironments associated with nucleic acids. PMID- 18406334 TI - Osteoblastic cells: differentiation and trans-differentiation. AB - The osteoblast is the bone forming cell and is derived from mesenchymal stem cells (MSC) present among the bone marrow stroma. MSC are capable of multi lineage differentiation into mesoderm-type cells such as osteoblasts and adipocytes. Understanding the mechanisms underlying osteoblast differentiation from MSC is a central topic in bone biology that can provide insight into mechanisms of bone maintenance and also novel pharmacological targets to increase osteoblast differentiation and consequently bone formation. PMID- 18406335 TI - An X-ray diffraction and X-ray absorption spectroscopy joint study of neuroglobin. AB - Neuroglobin (Ngb) is a member of the globin family expressed in the vertebrate brain, involved in neuroprotection. A combined approach of X-ray diffraction (XRD) on single crystal and X-ray absorption spectroscopy (XAS) in solution, allows to determine the oxidation state and the structure of the Fe-heme both in the bis-histidine and the CO-bound (NgbCO) states. The overall data demonstrate that under X-ray the iron is photoreduced fairly rapidly, and that the previously reported X-ray structure of ferric Ngb [B. Vallone, K. Nienhaus, M. Brunori, G.U. Nienhaus, Proteins 56 (2004) 85-92] very likely refers to a photoreduced species indistinguishable from the dithionite reduced protein. Results from the XAS analysis of NgbCO in solution are in good agreement with XRD data on the crystal. However prolonged X-ray exposure at 15K determines CO release. This preliminary result paves the way to experiments aimed at the characterization of pentacoordinate ferrous Ngb, the only species competent in binding external ligands such as O2, CO or NO. PMID- 18406337 TI - Ion channels and transporters in osteoclasts. AB - The resorbing osteoclast is an exceptional cell that secretes large amounts of acid through the coupled activity of a v-type H+-ATPase and a chloride channel that both reside in the ruffled membrane. Impairment of this acid secretion machinery by genetic mutations can abolish bone resorption activity, resulting in osteopetrotic phenotypes. Another key feature of osteoclasts is the transport of high amounts of calcium and phosphate from the resorption lacuna to the basolateral plasma membrane. Evidence exists that this occurs in part through entry of these ions into the osteoclast cytosol. Handling of such large amounts of a cellular messenger requires elaborate mechanisms. Membrane proteins that regulate osteoclast calcium homeostasis and the effect of calcium on osteoclast function and survival are therefore the second main focus of this review. PMID- 18406336 TI - Function, structure and regulation of the vacuolar (H+)-ATPases. AB - The vacuolar ATPases (or V-ATPases) are ATP-driven proton pumps that function to both acidify intracellular compartments and to transport protons across the plasma membrane. Intracellular V-ATPases function in such normal cellular processes as receptor-mediated endocytosis, intracellular membrane traffic, prohormone processing, protein degradation and neurotransmitter uptake, as well as in disease processes, including infection by influenza and other viruses and killing of cells by anthrax and diphtheria toxin. Plasma membrane V-ATPases are important in such physiological processes as urinary acidification, bone resorption and sperm maturation as well as in human diseases, including osteopetrosis, renal tubular acidosis and tumor metastasis. V-ATPases are large multi-subunit complexes composed of a peripheral domain (V(1)) responsible for hydrolysis of ATP and an integral domain (V(0)) that carries out proton transport. Proton transport is coupled to ATP hydrolysis by a rotary mechanism. V ATPase activity is regulated in vivo using a number of mechanisms, including reversible dissociation of the V(1) and V(0) domains, changes in coupling efficiency of proton transport and ATP hydrolysis and changes in pump density through reversible fusion of V-ATPase containing vesicles. V-ATPases are emerging as potential drug targets in treating a number of human diseases including osteoporosis and cancer. PMID- 18406338 TI - Osteoclast-osteoblast communication. AB - Cells in osteoclast and osteoblast lineages communicate with each other through cell-cell contact, diffusible paracrine factors and cell-bone matrix interaction. Osteoclast-osteoblast communication occurs in a basic multicellular unit (BMU) at the initiation, transition and termination phases of bone remodeling. At the initiation phase, hematopoietic precursors are recruited to the BMU. These precursors express cell surface receptors including c-Fms, RANK and costimulatory molecules, such as osteoclast-associated receptor (OSCAR), and differentiate into osteoclasts following cell-cell contact with osteoblasts, which express ligands. Subsequently, the transition from bone resorption to formation is mediated by osteoclast-derived 'coupling factors', which direct the differentiation and activation of osteoblasts in resorbed lacunae to refill it with new bone. Bidirectional signaling generated by interaction between ephrinB2 on osteoclasts and EphB4 on osteoblast precursors facilitates the transition. Such interaction is likely to occur between osteoclasts and lining cells in the bone remodeling compartment (BRC). At the termination phase, bone remodeling is completed by osteoblastic bone formation and mineralization of bone matrix. Here, we describe molecular communication between osteoclasts and osteoblasts at distinct phases of bone remodeling. PMID- 18406339 TI - Direct quantification of the four individual S states in Photosystem II using EPR spectroscopy. AB - EPR spectroscopy is very useful in studies of the oxygen evolving cycle in Photosystem II and EPR signals from the CaMn(4) cluster are known in all S states except S(4). Many signals are insufficiently understood and the S(0), S(1), and S(3) states have not yet been quantifiable through their EPR signals. Recently, split EPR signals, induced by illumination at liquid helium temperatures, have been reported in the S(0), S(1), and S(3) states. These split signals provide new spectral probes to the S state chemistry. We have studied the flash power dependence of the S state turnover in Photosystem II membranes by monitoring the split S(0), split S(1), split S(3) and S(2) state multiline EPR signals. We demonstrate that quantification of the S(1), S(3) and S(0) states, using the split EPR signals, is indeed possible in samples with mixed S state composition. The amplitudes of all three split EPR signals are linearly correlated to the concentration of the respective S state. We also show that the S(1) --> S(2) transition proceeds without misses following a saturating flash at 1 degrees C, whilst substantial misses occur in the S(2) --> S(3) transition following the second flash. PMID- 18406340 TI - Diseases caused by defects of mitochondrial carriers: a review. AB - A strikingly large number of mitochondrial DNA (mtDNA) mutations have been found to be the cause of respiratory chain and oxidative phosphorylation defects. These mitochondrial disorders were the first to be investigated after the small mtDNA had been sequenced in the 80s. Only recently numerous diseases resulting from mutations in nuclear genes encoding mitochondrial proteins have been characterized. Among these, nine are caused by defects of mitochondrial carriers, a family of nuclear-coded proteins that shuttle a variety of metabolites across the mitochondrial membrane. Mutations of mitochondrial carrier genes involved in mitochondrial functions other than oxidative phosphorylation are responsible for carnitine/acylcarnitine carrier deficiency, HHH syndrome, aspartate/glutamate isoform 2 deficiency, Amish microcephaly, and neonatal myoclonic epilepsy; these disorders are characterized by specific metabolic dysfunctions, depending on the physiological role of the affected carrier in intermediary metabolism. Defects of mitochondrial carriers that supply mitochondria with the substrates of oxidative phosphorylation, inorganic phosphate and ADP, are responsible for diseases characterized by defective energy production. Herein, all the mitochondrial carrier-associated diseases known to date are reviewed for the first time. Particular emphasis is given to the molecular basis and pathogenetic mechanism of these inherited disorders. PMID- 18406341 TI - FTIR studies show lipophilic moisturizers to interact with stratum corneum lipids, rendering the more densely packed. AB - Lipophilic moisturizers are widely used to treat dry skin. However, their interaction with the lipids in the upper layer of the skin, the stratum corneum (SC), is largely unknown. In the present study this interaction of three moisturizers, isostearyl isostearate (ISIS), isopropyl isostearate (IPIS) and glycerol monoisostearate (GMIS), has been elucidated using lipid mixtures containing isolated ceramides (CER), cholesterol (CHOL) and free fatty acids (FFA), mimicking the lipid composition and organization in SC. The conformational ordering and the lateral packing of the lipid mixtures were examined by Fourier transformed infrared spectroscopy. Equimolar CER:CHOL:FFA mixtures show an orthorhombic to hexagonal phase transition between 22 and 30 degrees C and an ordered-disordered phase transition between 46 and 64 degrees C. Addition of 20% m/m ISIS or IPIS increased the thermotropic stability of the orthorhombic lateral packing, while GMIS had no influence. Furthermore, small amounts of all three moisturizers are incorporated into the CER:CHOL:FFA lattice, while the majority of the moisturizer exists in separate domains. Especially the thermotropic stabilization of the orthorhombic lateral packing, which might reduce water loss from the skin, is considered to contribute to the moisturizing effect of IPIS and ISIS in stratum corneum. PMID- 18406342 TI - Towards a systems biology approach to study type II/IV secretion systems. AB - Many gram-negative bacteria produce thin protein filaments, named pili, which extend beyond the confines of the outer membrane. The importance of these pili is illustrated by the fact that highly complex, multi-protein pilus-assembly machines have evolved, not once, but several times. Their many functions include motility, adhesion, secretion, and DNA transfer, all of which can contribute to the virulence of bacterial pathogens or to the spread of virulence factors by horizontal gene transfer. The medical importance has stimulated extensive biochemical and genetic studies but the assembly and function of pili remains an enigma. It is clear that progress in this field requires a more holistic approach where the entire molecular apparatus that forms the pilus is studied as a system. In recent years systems biology approaches have started to complement classical studies of pili and their assembly. Moreover, continued progress in structural biology is building a picture of the components that make up the assembly machine. However, the complexity and multiple-membrane spanning nature of these secretion systems pose formidable technical challenges, and it will require a concerted effort before we can create comprehensive and predictive models of these remarkable molecular machines. PMID- 18406343 TI - Synthetic retinoid CD437 induces mitochondria-mediated apoptosis in hepatocellular carcinoma cells. AB - Retinoids play an important role in the regulation of cell growth and death. Synthetic retinoid CD437 reportedly induces apoptosis in various cancer cell lines. However, the mechanism of inducing apoptosis in hepatocellular carcinoma (HCC) cells by this agent remains to be clarified. In this study, we investigated the signaling pathway by which CD437 induces apoptosis in HCC cell lines. Apoptosis of six human HCC cell lines was induced by treatment with CD437. Caspase-3 and -9 were activated by CD437, suggesting that the apoptosis is mediated by mitochondrial pathways. Consistent with these findings, the treatment with CD437 upregulated Bax protein, downregulated Bcl-2 protein and released cytochrome c into the cytoplasm. Moreover, rhodamine123 staining revealed mitochondrial depolarization in the cells treated with CD437. These data of the present study suggest that CD437 induces apoptosis in HCC cells via mitochondrial pathways. PMID- 18406344 TI - A novel role for thioredoxin reductase in the iron metabolism of S. cerevisiae. AB - Intracellular levels of iron are tightly regulated. Saccharomyces cerevisiae uses well-defined pathways to extract iron molecules from the environment. Once inside the cell, the iron molecules must be transferred to target sites via an intracellular iron transporter. Although analogous carriers have been described for other metals, such as copper, an iron transporter has yet to be identified. We used two-dimensional gel electrophoresis and mass spectrometry techniques to attempt to identify the iron transporter from cytosolic fraction of S. cerevisiae. In this study, we identified the iron-binding activity of thioredoxin reductase, and our data suggest a potential role for this enzyme in intracellular iron transport. PMID- 18406345 TI - New efficient replication system with hepatitis C virus genome derived from a patient with acute hepatitis C. AB - We report for the first time a new RNA replication system with a hepatitis C virus (HCV) strain (AH1) derived from a patient with acute hepatitis C. Using an HCV replicon RNA library constructed with the AH1 strain (genotype 1b), we first established a cloned cell line, sAH1, harboring the HCV replicon. Cured cells obtained with interferon treatment of sAH1 cells were used for transfection with genome-length HCV RNA possessing four mutations found in sAH1 replicon. Consequently, one cloned cell line, AH1, supporting efficient replication of genome-length HCV RNA was obtained. By the comparison of AH1 cells with the O cells supporting genome-length HCV RNA (HCV-O strain) replication, we found different anti-HCV profiles of interferon-gamma and cyclosporine A between AH1 and O cells. Reporter assay analysis suggests that the diverse effects of interferon-gamma are due to the difference in HCV strains, but not the cellular environment. PMID- 18406346 TI - MAP kinase: it's been longer than fifteen minutes. AB - The review highlights evidence for different functions in the cell cycle of the two MAP kinase kinases, MEK1 and MEK2, and the two MAP kinases, ERK1 and ERK2. Functional differences may explain why instances of cell cycle arrest can be MEK1 or MEK2 dependent. PMID- 18406347 TI - AM251 suppresses the viability of HepG2 cells through the AMPK (AMP-activated protein kinase)-JNK (c-Jun N-terminal kinase)-ATF3 (activating transcription factor 3) pathway. AB - AM251, a cannabinoid antagonist, has various biological activities. In this study, we found that AM251 suppressed the viability of hepatoma HepG2 cells and also increased phosphorylation of JNK (c-jun N-terminal kinase) and ATF3 (activating transcription factor 3). In addition, AM251 phosphorylated AMPK (AMP activated protein kinase) in a time and dose-dependent manner. Inhibition of AMPK blocked AM251-induced JNK/ATF3 phosphorylation. Expression of AMPK or treatment with AICAR (5-aminoimidazole-4-carboxy-amide-1-d-ribofuranoside), an AMPK activator, activated the JNK/ATF3 pathways. Together, these results suggest that AM251 may have anti-tumor effects in hepatoma through activation of the AMPK-JNK ATF3 signal pathway. PMID- 18406348 TI - Curcumin inhibits bTREK-1 K+ channels and stimulates cortisol secretion from adrenocortical cells. AB - Bovine adrenal zona fasciculata (AZF) cells express bTREK-1 K(+) channels that set the resting membrane potential. Inhibition of these channels by adrenocorticotropic hormone (ACTH) is coupled to membrane depolarization and cortisol secretion. Curcumin, a phytochemical with medicinal properties extracted from the spice turmeric, was found to modulate both bTREK-1 K(+) currents and cortisol secretion from AZF cells. In whole-cell patch clamp experiments, curcumin inhibited bTREK-1 with an IC(50) of 0.93muM by a mechanism that was voltage-independent. bTREK-1 inhibition by curcumin occurred through interaction with an external binding site and was independent of ATP hydrolysis. Curcumin produced a concentration-dependent increase in cortisol secretion that persisted for up to 24h. At a maximally effective concentration of 50muM, curcumin increased secretion as much as 10-fold. These results demonstrate that curcumin potently inhibits bTREK-1 K(+) channels and stimulates cortisol secretion from bovine AZF cells. The inhibition of bTREK-1 by curcumin may be linked to cortisol secretion through membrane depolarization. Since TREK-1 is widely expressed in a variety of cells, it is likely that some of the biological actions of curcumin, including its therapeutic effects, may be mediated through inhibition of these K(+) channels. PMID- 18406350 TI - Secreted proprotein convertase subtilisin/kexin type 9 reduces both hepatic and extrahepatic low-density lipoprotein receptors in vivo. AB - Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a serine protease that is known to reduce hepatic low-density lipoprotein receptor (LDLR) levels and increase plasma LDL cholesterol. It is not clear, however, whether secreted PCSK9 degrades extrahepatic LDLRs. We present evidence that recombinant PCSK9, either injected intravenously into or expressed in the liver of C57BL/6 mice, significantly reduced LDLR levels in multiple extrahepatic tissues. During the initial characterization, we found that injected human recombinant PCSK9 at 30 microg/mouse had a half-life of 15 min in serum in mice. Hepatic LDLR levels were reduced within 30min and the degradation of hepatic LDLR reached the maximum 2h after the initial protein injection. Endocytosis of PCSK9 in liver occurred within 5min of protein injection and internalized PCSK9 was only barely detectable within 1h. When extrahepatic LDLRs were examined by Western blotting analysis, we found significant reductions of LDLRs in multiple extrahepatic tissues including lung, adipose and kidney along with the more dramatic reduction of LDLRs in liver. These studies were further extended using adenoviral expression of human PCSK9 in C57BL/6 mice to demonstrate that PCSK9 produced in liver impacted extrahepatic tissue LDLR levels as well. Taken together, our studies indicate that secreted PCSK9 can potentially impact extrahepatic tissue cholesterol homeostasis by regulating extrahepatic tissue LDLR levels. PMID- 18406349 TI - Interferon alfacon 1 inhibits SARS-CoV infection in human bronchial epithelial Calu-3 cells. AB - The primary targets for SARS-CoV infection are the epithelial cells in the respiratory and intestinal tract. The angiotensin-converting enzyme 2 (ACE-2) has been identified as a functional receptor for SARS-CoV. ACE-2 has been shown to be expressed at the apical domain of polarized Calu-3 cells. In this report, interferon alfacon 1 was examined for inhibitory activities against SARS-CoV on human lung carcinoma epithelial Calu-3 cell line and the other three African green monkey kidney epithelial cell lines. Interferon alfacon 1 demonstrated significant antiviral activity in neutral red uptake assay and virus yield reduction assay. The data might provide an important insight into the mechanism of pathogenesis of SARS-CoV allowing further development of antiviral therapies for treating SARS infections. PMID- 18406351 TI - Insulin-secreting L-cells for the treatment of insulin-dependent diabetes. AB - Cell-based treatments for insulin-dependent diabetes (IDD) may provide more physiologic regulation of blood glucose levels than daily insulin injections, thereby reducing the occurrence of secondary complications associated with diabetes. An autologous cell source is especially attractive for regulatory and ethical reasons in addition to eliminating the need for immunosuppression. This study uses non-beta-cells, genetically modified for physiologic insulin secretion. Enteroendocrine L-cells, exhibit regulated secretion in response to physiologic stimuli and their endogenous products are fully compatible with prandial metabolism. Murine GLUTag L-cells were transfected with a plasmid co expressing human insulin and neomycin resistance and the stable cell line, GLUTag INS, was established. Secretion properties of GLUTag-INS cells were investigated in vitro through induced secretion tests using meat hydrolysate or 3-isobutyl-1 methylxanthine and forskolin as secretagogues. GLUTag-INS cells rapidly co secreted recombinant insulin and endogenous glucagon-like peptide in response to metabolic cues from the surrounding medium and demonstrated efficient processing of proinsulin to insulin. PMID- 18406352 TI - Measurement of free concentrations of highly protein-bound warfarin in plasma by ultra performance liquid chromatography-tandem mass spectrometry and its correlation with the international normalized ratio. AB - BACKGROUND: A novel method has been developed for the determination of free warfarin in human plasma by ultra performance liquid chromatography combined with tandem mass spectrometry (UPLC-MS/MS). METHODS: The plasma filtrate was prepared by a high-speed ultrafiltration (UF), and was then extracted by a liquid/liquid extraction (LLE) with ethyl acetate. The chromatographic separation was performed on an Acquity UPLC BEH C(18) (2.1 mm x 50 mm, 1.7 microm, Waters) with a mobile phase consisting of 0.1% formic acid/acetonitrile (50:50, v/v). The analyses were carried out by multiple reaction monitoring (MRM) using the precursor-to-product combinations of m/z 307.3-->161.2 for warfarin and m/z 347.2-->161.3 for Cl-War. RESULTS: The lower limit of quantification (LLOQ) was 0.25 ng/ml and the assay exhibited a linear range of 0.25-16 ng/ml and gave a correlation coefficient (r) of > or = 0.9999. Quality control samples (0.5, 2, 8 ng/ml) in 5 replicates from 3 different runs of analysis demonstrated intra-assay CVs of 3.0-10.2%, inter assay CVs of 4.2-12.0%, and an overall accuracy of 85-115%. CONCLUSIONS: The method can be applied to analyze the correlation of free concentration of warfarin and the international normalized ratio (INR). PMID- 18406353 TI - Downregulation of CCND1 and CDK6 by miR-34a induces cell cycle arrest. AB - miRNAs regulate gene expression by inhibiting translation or by targeting messenger RNA (mRNA) for degradation in a post-transcriptional fashion. In the present study, we show that ectopic expression of miR-34a reduces both mRNA and protein levels of cyclin D1 (CCND1) and cyclin-dependent kinase 6 (CDK6). We also demonstrate that miR-34a targets the 3'-untranslated mRNA region of CCND1 as well as CDK6, which in turn interferes with phosphorylation of retinoblastoma. In addition, we show that overexpression of miR-34a induces a significant G1 cell cycle arrest in the A549 cell line. Taken together, our data suggest that the effects of miR-34a on G1 cell cycle arrest are through the down-regulation of CCND1 and CDK6, which is associated with other targets of miR-34a either additively or synergistically. PMID- 18406354 TI - Monitoring fibril formation of the N-terminal domain of PABPN1 carrying an alanine repeat by tryptophan fluorescence and real-time NMR. AB - Intranuclear fibrils due to poly-alanine expansions in the N-terminal domain of the poly(A) binding protein PABPN1 correlate with the disease oculopharyngeal muscular dystrophy (OPMD). For monitoring fibril formation by fluorescence and real-time NMR spectroscopy, tryptophans were introduced either into the middle or C-terminal of the poly-alanine segment. The kinetics of fibril formation which were monitored by fluorescence spectroscopy were matched by real-time NMR kinetics. Our results show that fibril formation is concomitant with the burial of the tryptophans in the fibrillar core. Since no soluble pre-fibrillar intermediate(s) was detected, fibril formation of this domain may be regarded as a two state conversion from an unfolded soluble into folded insoluble species. PMID- 18406355 TI - Inactivation of epoxide hydrolase by catalysis-induced formation of isoaspartate. AB - Epoxide hydrolases catalyze hydrolytic epoxide ring-opening, most often via formation of a covalent hydroxyalkyl-enzyme intermediate. A mutant of Agrobacterium radiobacter epoxide hydrolase, in which the phenylalanine residue that flanks the invariant catalytic aspartate nucleophile is replaced by a threonine, exhibited inactivation during conversion when the (R)-enantiomer of para-nitrostyrene epoxide was used as substrate. HPLC analysis of tryptic fragments of the epoxide hydrolase, followed by MALDI-TOF and TOF/TOF analysis, indicated that inactivation was due to conversion of the nucleophilic aspartate into isoaspartate, which represents a novel mechanism of catalysis-induced autoinactivation. Inactivation occurred at a lower rate with the (S)-enantiomer of para-nitrostyrene epoxide, indicating that it is related to the structure of the covalent hydroxyalkyl-enzyme intermediate. PMID- 18406356 TI - Negative regulation of apoptosis in yeast. AB - In recent years, yeast has been proven to be a useful model organism for studying programmed cell death. It not only exhibits characteristic markers of apoptotic cell death when heterologous inducers of apoptosis are expressed or when treated with apoptosis inducing drugs such as hydrogen peroxide (H(2)O(2)) or acetic acid, but contains homologues of several components of the apoptotic machinery identified in mammals, flies and nematodes, such as caspases, apoptosis inducing factor (AIF), Omi/HtrA2 and inhibitor-of-apoptosis proteins (IAPs). In this review, we focus on the role of negative regulators of apoptosis in yeasts. Bir1p is the only IAP protein in Saccharomyces cerevisiae and has long been known to play a role in cell cycle progression by acting as kinetochore and chromosomal passenger protein. Recent data established Bir1p's protective function against programmed cell death induced by H(2)O(2) treatment and in chronological ageing. Other factors that have a direct or indirect influence on intracellular levels of reactive oxygen species (ROS) and thus lead to apoptosis if they are misregulated or non-functional will be discussed. PMID- 18406358 TI - Mitochondria-dependent apoptosis in yeast. AB - Mitochondrial involvement in yeast apoptosis is probably the most unifying feature in the field. Reports proposing a role for mitochondria in yeast apoptosis present evidence ranging from the simple observation of ROS accumulation in the cell to the identification of mitochondrial proteins mediating cell death. Although yeast is unarguably a simple model it reveals an elaborate regulation of the death process involving distinct proteins and most likely different pathways, depending on the insult, growth conditions and cell metabolism. This complexity may be due to the interplay between the death pathways and the major signalling routes in the cell, contributing to a whole integrated response. The elucidation of these pathways in yeast has been a valuable help in understanding the intricate mechanisms of cell death in higher eukaryotes, and of severe human diseases associated with mitochondria-dependent apoptosis. In addition, the absence of obvious orthologues of mammalian apoptotic regulators, namely of the Bcl-2 family, favours the use of yeast to assess the function of such proteins. In conclusion, yeast with its distinctive ability to survive without respiration-competent mitochondria is a powerful model to study the involvement of mitochondria and mitochondria interacting proteins in cell death. PMID- 18406357 TI - Differential regulation of Kruppel-like factor family transcription factor expression in neonatal rat cardiac myocytes: effects of endothelin-1, oxidative stress and cytokines. AB - Kruppel-like transcription factors (Klfs) modulate fundamental cell processes. Cardiac myocytes are terminally-differentiated, but hypertrophy in response to stimuli such as endothelin-1. H2O2 or cytokines promote myocyte apoptosis. Microarray studies of neonatal rat myocytes identified several Klfs as endothelin 1-responsive genes. We used quantitative PCR for further analysis of Klf expression in neonatal rat myocytes. In response to endothelin-1, Klf2 mRNA expression was rapidly increased ( approximately 9-fold; 15-30 min) with later increases in expression of Klf4 and Klf6 ( approximately 5-fold; 30-60 min). All were regulated as immediate early genes (cycloheximide did not inhibit the increases in expression). Klf5 expression was increased at 1-2 h ( approximately 13-fold) as a second phase response (cycloheximide inhibited the increase). These increases were transient and attenuated by U0126. H2O2 increased expression of Klf2, Klf4 and Klf6, but interleukin-1beta or tumor necrosis factor alpha downregulated Klf2 expression with no effect on Klf4 or Klf6. Of the Klfs which repress transcription, endothelin-1 rapidly downregulated expression of Klf3, Klf11 and Klf15. The dynamic regulation of expression of multiple Klf family members in cardiac myocytes suggests that, as a family, they are actively involved in regulating phenotypic responses (hypertrophy and apoptosis) to extracellular stimuli. PMID- 18406359 TI - Novel translocation responses of cytosolic phospholipase A2alpha fluorescent proteins. AB - Cytosolic phospholipase A2 (cPLA2)alpha responds to the rise in cytosolic Ca2+ ([Ca2+]i) attending cell stimulation by moving to intracellular membranes, releasing arachidonic acid (AA) from these membranes, and thereby initiating the synthesis of various lipid mediators. Under some conditions, however, cPLA2alpha translocation occurs without any corresponding changes in [Ca2+]i. The signal for such responses has not been identified. Using confocal microscopy to track fluorescent proteins fused to cPLA2alpha or cPLA2alpha's C2 domain, we find that AA mimics Ca2+ ionophores in stimulating cPLA(2)alpha translocations to the perinuclear ER and to a novel site, the lipid body. Unlike the ionophores, AA acted independently of [Ca2+](i) rises and did not translocate the proteins to the Golgi. AA's action did not involve its metabolism to eicosanoids or acylation into cellular lipids. Receptor agonists also stimulated translocations targeting lipid bodies. We propose that AA is a signal for Ca2+-independent cPLA2alpha translocation and that lipid bodies are common targets of cPLA2alpha and contributors to stimulus-induced lipid mediator synthesis. PMID- 18406360 TI - Cholesterol is a determinant of the structures of discoidal high density lipoproteins formed by the solubilization of phospholipid membranes by apolipoprotein A-I. AB - Formation of discoidal high density lipoproteins (rHDL) by apolipoprotein A-I (apoA-I) mediated solubilization of dimyristoyl phosphatidylcholine (DMPC) multilamellar vesicles (MLV) was dramatically affected by bilayer cholesterol concentration. At a low ratio of DMPC/apoA-I (2 mg DMPC/mg apoA-I, 84/1 mol/mol), sterols (cholesterol, lathosterol, and beta-sitosterol) that form ordered lipid phases increase the rate of solubilization similarly, yielding rHDL with similar structures. By changing the temperature and sterol concentration, the rates of solubilization varied almost 3 orders of magnitude; however, the sizes of the rHDL were independent of the rate of their formation and dependent upon the bilayer sterol concentration. At a high ratio of DMPC/apoA-I (10/1 mg DMPC/mg apoA-I, 420/1 mol/mol), changing the temperature and cholesterol concentration yielded rHDL that varied greatly in size, phospholipid/protein ratio, mol% cholesterol, and number of apoA-I molecules per particle. rHDL were isolated that had 2, 4, 6, and 8 molecules of apoA-I per particle, mean diameters of 117, 200, 303, and 396 A, and a mol% cholesterol that was similar to the original MLV. Kinetic studies demonstrated that the different sized rHDL are formed independently and concurrently. The rate of formation, lipid composition, and three-dimensional structures of cholesterol-rich rHDL is dictated primarily by the original membrane phase properties and cholesterol content. The size speciation of rHDL and probably nascent HDL formed via the activity of the ABCA1 lipid transporter is mechanistically linked to the cholesterol content of the membranes from which they were formed. PMID- 18406362 TI - [Abstracts from the French-speaking Meeting of Neurology, 23-26 April 2008, Bordeaux, France]. PMID- 18406363 TI - Innate immunity. PMID- 18406364 TI - Transcriptional responses of Arabidopsis thaliana to the bacteria-derived PAMPs harpin and lipopolysaccharide. AB - Many plant-pathogen interactions are controlled by specific interactions between pathogen avirulence (avr) gene loci and the corresponding plant resistance R locus (gene-for-gene-hypothesis). Very often, this type of interaction culminates in a hypersensitive reaction (HR). However, recently pathogen-associated molecular patterns (PAMPs) such as flagellin or lipopolysaccharides (LPS) that are common to all bacteria have been shown to act as general elicitors of basal or innate immune responses in several plant species. Here, we summarize the genetic programs in Arabidopsis thaliana behind the LPS-induced basal response and the HR induced by harpin, respectively. Using Agilent Arabidopsis cDNA microarrays consisting of approximately 15,000 oligomers, changes in transcript accumulation of treated cells were monitored over a period of 24h after elicitor treatment. Analysis of the array data revealed significant responses to LPS (309 genes), harpin (951 genes) or both (313 genes). Concentrating our analysis on the genes encoding transcription factors, defence genes, cell wall biogenesis-related genes and signal transduction components we monitored interesting parallels, but also remarkably different expression patterns. Harpin and LPS induced an overlapping set of genes involved in cell wall biogenesis, cellular communication and signalling. The pattern of induced genes associated with cell rescue and general stress responses such as small heat-shock proteins was highly similar. In contrast, there is a striking difference regarding some of the most prominent, central components of plant defence such as WRKY transcription factors and oxidative burst-associated genes like NADPH oxidases, whose expression became apparent only after treatment with harpin. While both harpin and LPS can stimulate plant immunity in Arabidopsis, the PAMP LPS induces much more subtle host reactions at the transcriptome scale. The defence machinery induced by harpin resembles the known HR-type host responses leading to cell death after treatment with this elicitor. LPS is a weak inducer of basal resistance and induces a different pattern of genes. Strikingly the biggest overlap (40) of responding genes was found between the early harpin response (30min) and the late LPS response (24h). PMID- 18406365 TI - Innate immune functions of human gammadelta T cells. AB - gammadelta T cells expressing the Vgamma9Vdelta2 T cell receptor (TCR) account for 1-10% of CD3(+) peripheral blood T lymphocytes. Vgamma9Vdelta2 T cells use their TCR as a pattern recognition receptor to sense the presence of infection through specific recognition of intermediates of the microbial non-mevalonate pathway of isoprenoid biosynthesis. Such phosphoantigens rapidly and selectively activate human gammadelta T cells to produce proinflammatory cytokines, notably interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). In addition, human gammadelta T cells express certain Toll-like receptors (TLR) and directly respond to the corresponding ligands. We have demonstrated expression of TLR3 in Vgamma9Vdelta2 T cells and striking costimulatory effects of the ligand polyinosinic-polycytidylic acid (polyI:C) on TCR-stimulated IFN-gamma production. Gene expression studies by microarray analysis identified additional genes that were up-regulated by combined TCR- and TLR3 stimulation. We discuss these findings in the context of the suspected role of human Vgamma9Vdelta2 T cells as a link between innate and adaptive immune responses. PMID- 18406361 TI - Lipid peroxidation and decomposition--conflicting roles in plaque vulnerability and stability. AB - The low density lipoprotein (LDL) oxidation hypothesis has generated considerable interest in oxidative stress and how it might affect atherosclerosis. However, the failure of antioxidants, particularly vitamin E, to affect the progression of the disease in humans has convinced even staunch supporters of the hypothesis to take a step backwards and reconsider alternatives. Preponderant evidence for the hypothesis came from animal antioxidant intervention studies. In this review we point out basic differences between animal and human atherosclerosis development and suggest that human disease starts where animal studies end. While initial oxidative steps in the generation of early fatty streak lesions might be common, the differences might be in the steps involved in the decomposition of peroxidized lipids into aldehydes and their further oxidation into carboxylic acids. We suggest that these steps may not be amenable to attenuation by antioxidants and antioxidants might actually counter the stabilization of plaque by preventing the formation of carboxylic acids which are anti-inflammatory in nature. The formation of such dicarboxylic acids may also be conducive to plaque stabilization by trapping calcium. We suggest that agents that would prevent the decomposition of lipid peroxides and promote the formation and removal of lipid hydroxides, such as paraoxonase (PON 1) or apo A1/high density lipoprotein (HDL) might be more conducive to plaque regression. PMID- 18406366 TI - Neutrophil granulocytes as host cells and transport vehicles for intracellular pathogens: apoptosis as infection-promoting factor. AB - Polymorphonuclear neutrophil granulocytes (PMN) are primary antimicrobial effector cells of the innate immune system and serve to destroy invading pathogens. Although most ingested microorganisms are killed readily inside PMN, several obligate or facultative intracellular pathogens survive even in this hostile environment. Extension of the life span of neutrophils is a general escape mechanism of pathogens residing in PMN. However, after 2-4 days, even infected neutrophils become apoptotic and are phagocytosed by macrophages. Since microbes entering macrophages via the uptake of infected apoptotic PMN may survive and multiply in macrophages, apoptotic neutrophils can serve as "Trojan horses" for certain pathogens. Interfering with activating signaling pathways appears to be another potent mechanism by which intracellular microorganisms suppress cellular activation in neutrophils. In addition to provide a short overview of the topic, the present review aims to summarize our own findings regarding the interaction between human neutrophils and intracellular pathogens as well as regarding the disease promoting role of apoptotic cells after infection with Leishmania major. PMID- 18406367 TI - Lipopolysaccharide sensing an important factor in the innate immune response to Gram-negative bacterial infections: benefits and hazards of LPS hypersensitivity. AB - In this review, we summarize our investigations concerning the differential importance of CD14 and LBP in toll-like receptor 4 (TLR4)/myeloid differentiation protein-2 (MD-2)-mediated signaling by smooth and rough-form lipopolysaccharide (LPS) chemotypes and include the results obtained in studies with murine and human TLR4-transgenic mice. Furthermore, we present more recent data on the mechanisms involved in the induction of LPS hypersensitivity by bacterial and viral infections and on the reactivity of the hypersensitive host to non-LPS microbial ligands and endogenous mediators. Finally, the effects of pre-existing hypersensitivity on the course and outcome of a super-infection with Salmonella typhimurium or Listeria monocytogenes are summarized. PMID- 18406368 TI - TLR2 - promiscuous or specific? A critical re-evaluation of a receptor expressing apparent broad specificity. AB - Of all pattern recognition receptors (PRR) in innate immunity, Toll-like receptor 2 (TLR2) recognizes the structurally broadest range of different bacterial compounds known as pathogen-associated molecular patterns (PAMPs). TLR2 agonists identified so far are lipopolysaccharides (LPSs) from different bacterial strains, lipoproteins, (synthetic) lipopeptides, lipoarabinomannans, lipomannans, glycosylphosphatidylinositol, lipoteichoic acids (LTA), various proteins including lipoproteins and glycoproteins, zymosan, and peptidoglycan (PG). Because these molecules are structurally diverse, it seems unlikely that TLR2 has the capability to react with all agonists to the same degree. The aim of this review is to identify and describe well-defined structure-function relationships for TLR2. Because of its biomedical importance and because its genetics and biochemistry are presently most completely known among all Gram-positive bacteria, we have chosen Staphylococcus aureus as a focus. Our data together with those reported by other groups reveal that only lipoproteins/lipopeptides are sensed at physiologically concentrations by TLR2 at picomolar levels. This finding implies that the activity of all other putative bacterial compounds so far reported as TLR2 agonists was most likely due to contaminating highly active natural lipoproteins and/or lipopeptides. PMID- 18406369 TI - Regulation of innate immunity by suppressor of cytokine signaling (SOCS) proteins. AB - Innate immunity represents the first line of defense against invading pathogens. Toll-like receptors (TLRs) are important for activation of innate immunity. Moreover, cytokines mediate communication of cells and are necessary to mount an appropriately regulated immune response. However, activation of innate immunity has to be tightly controlled to avoid overshooting immune reactions. Suppressor of cytokine signaling (SOCS) proteins have been identified as inducible feedback inhibitors of cytokine receptors and have been shown to be of crucial importance for the limitation of inflammatory responses. In this review, we describe the role of SOCS proteins in macrophages and dendritic cells (DCs). Based on our own findings, we show that SOCS proteins are directly induced by stimulation of TLRs. However, SOCS proteins do not interfere with direct TLR signaling, but avoid overshooting activation by regulating paracrine IFN-beta signaling. In addition, SOCS proteins in macrophages and DCs regulate the sensitivity towards IFN-gamma and GM-CSF, thereby modulating anti-microbial activity of macrophages and differentiation of DCs. We discuss that SOCS induction can also be used by microbes to evade immune defense, and this is exemplified by the parasite Toxoplasma gondii which induces SOCS1 to inhibit IFN-gamma-mediated macrophage activation. Taken together, the findings indicate that SOCS proteins play an important role in the balanced activation of innate immunity during infectious encounter. PMID- 18406370 TI - Specificity of the innate immune system and diversity of C-type lectin domain (CTLD) proteins in the nematode Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans has become an important model for the study of innate immunity. Its immune system is based on several signaling cascades, including a Toll-like receptor, three mitogen-activated protein kinases (MAPK), one transforming growth factor-beta (TGF-beta), the insulin-like receptor (ILR), and the programmed cell death (PCD) pathway. Furthermore, it also involves C-type lectin domain- (CTLD) containing proteins as well as several classes of antimicrobial effectors such as lysozymes. Almost all components of the nematode immune system have homologs in other organisms, including humans, and are therefore likely of ancient evolutionary origin. At the same time, most of them are part of a general stress response, suggesting that they only provide unspecific defense. In the current article, we re-evaluate this suggestion and explore the level of specificity in C. elegans innate immunity, i.e. the nematode's ability to mount a distinct defense response towards different pathogens. We draw particular attention to the CTLD proteins, which are abundant in the nematode genome (278 genes) and many of which show a pathogen-specific response during infection. Specificity may also be achieved through the differential activation of antimicrobial genes, distinct functions of the immunity signaling cascades as well as signal integration across pathways. Taken together, our evaluation reveals high potential for immune specificity in C. elegans that may enhance the nematode's ability to fight off pathogens. PMID- 18406371 TI - Mast cell functions in the innate skin immune system. AB - Mast cells are not only potent effector cells in allergy, but are also important players in protective immune responses against pathogens. Most of our knowledge about mast cells in innate immunity is derived from models of sepsis, whereas their role in innate immune responses of the skin has largely been neglected in the past. Their particular pattern of distribution in the skin and their ability to sense and react to pathogens and other danger signals indicate that mast cells can be important sentinels and effector cells in skin immune responses. The recent findings reviewed here have confirmed this hypothesis and have established a prominent role for skin mast cells in innate immunity. PMID- 18406372 TI - Crosstalk of signalling processes of innate immunity with Yersinia Yop effector functions. AB - The interaction of microbial pathogens with host cells critically determines the genesis of infectious diseases. Gram-negative, pathogenic bacteria from the genus Yersinia deliver a set of virulence proteins, the so-called Yersinia outer proteins (Yops), inside the eukaryotic cell where the Yops perturb key cellular functions of innate immunity. In our past work, we used Yersinia enterocolitica as a tool to explore the crosstalk between the bacterial pathogen and its host cell. Yersiniae counteract phagocytosis, suppress proinflammatory signalling and trigger apoptosis in macrophages. Macrophage cell death results from the deregulation of Toll-like receptors-dependent conserved signalling pathways by Yersinia infection. We summarize our current understanding about the signals and reactions elicited on both the bacterial and host cell sides that determine the fate of the infected cell along with the innate immune response. PMID- 18406373 TI - New aspects of NK cell subset identification and inference of NK cells' regulatory capacity by assessing functional and genomic profiles. AB - Natural killer (NK) cells represent important early effector cells of the innate immune system. They can lyse virally infected and malignant cells without prior sensitization, making them important members of the first line of defense. In addition, they participate in the regulation of immune responses and hematopoiesis by producing various cytokines and chemokines. These different functional capacities can to some extent be assigned to distinct NK cell populations. In humans, CD56(dim) NK cells featuring strong cytotoxic capacity can be easily distinguished from CD56(bright) NK cells, which predominantly produce cytokines and exert only marginal cytotoxicity. Murine NK cells lack CD56 expression and no correlate marker enabling identification of functional subpopulations has been described. Here, we summarize and discuss human NK cell populations in greater detail in order to evaluate their regulatory capacity and to detect alternative and distinctive markers, e.g. CXCR3 and CD27, which are shared by both humans and mice. PMID- 18406374 TI - Cytokine induction by Gram-positive bacteria. AB - Despite similar clinical relevance of Gram-positive and Gram-negative infections, immune activation by Gram-positive bacteria is by far less well understood than immune activation by Gram-negative bacteria. Our group has made available highly purified lipoteichoic acids (LTA) as a key Gram-positive immunostimulatory component. We have characterized the reasons for lower potency of LTA compared to Gram-negative lipopolysaccharide (LPS), identifying lack of IL-12/IFNgamma induction as a general characteristic of TLR2 agonists, and need for presentation of LTA on surfaces for enhanced immunostimulatory potency, as major aspects. Aspects of chemokine induction, where LTA is more potent than LPS, have been addressed. Furthermore, novel complement and plant defence activation, as well as CD36 as a new LTA receptor, were identified. The bacterial costimuli and modulators of LTA inducible responses are being investigated: LTA isolated from so far 16 bacterial species, although different in structure, behave remarkably similar while whole live and killed bacteria differ with regard to the pattern of induced responses. The purification and characterization of the respective components of the bacterial cell wall has begun. PMID- 18406375 TI - Ficolins: novel pattern recognition molecules of the innate immune response. AB - Ficolins are members of the collectin family of proteins which are able to recognize pathogen-associated molecular pattern (PAMP) on microbial surfaces. Upon binding to their specific PAMP, ficolins may trigger activation of the immune system by either binding to cellular receptors for collectins or by initiating activation of complement via the lectin pathway. For the latter, the human ficolins (i.e. L-, H- and M-ficolin) and murine ficolin-A were shown to associate with the lectin pathway-specific serine protease MBL-associated serine protease-2 (MASP-2) and catalyse its activation which in turn activates C4 and C4b-bound C2 to generate the C3 convertase C4b2a. There is mounting evidence underlining the lectin nature of ficolins with a wide range of carbohydrate moieties recognized on microbial surfaces. However, not all members of the ficolin family appear to act as lectin pathway recognition components. For example, murine ficolin-B does not associate with MASP-2 and appears to be absent in plasma and other humoral fluids. Its stringent cellular localization points to other functions within the immune response, possibly acting as an intracellular scavenger to target and facilitate clearance of PAMP-bearing debris. When comparing ficolin orthologues from different species, it appears evident that human, murine, and porcine ficolins differ in many aspects, a specific point that we aim to address in this review. PMID- 18406376 TI - Fusogenicity of membranes: the impact of acid sphingomyelinase on innate immune responses. AB - Acid sphingomyelinase (ASMase) has been implemented in cellular signaling mainly because its reaction product, ceramide, has been assumed to be a mediator within signaling pathways. Our studies of three independent infection systems show that ASMase is required for phago-lysosomal fusion in macrophages infected with Listeria monocytogenes, for exocytosis of secretory lysosomes by lymphocytic choriomeningitis virus-specific cytotoxic T cells, and for generation of multinucleated giant cells in granuloma of mice infected with Mycobacterium avium. Because of its neutral lipid nature, ceramide is confined to the membranes of phagosomes and lysosomes or the extracellular leaflet of the plasma membrane. In light of the biochemical and biophysical properties of ceramide, we provide a model suggesting that ASMase regulates select vesicular fusion processes by modifying the steric conformation of cellular membranes. PMID- 18406377 TI - Recognition of nucleic acid and nucleic acid analogs by Toll-like receptors 7, 8 and 9. AB - The mammalian immune system senses pathogens through pattern recognition receptors (PRR) and responds with activation. Toll-like receptors (TLRs) that are expressed on immune and non-immune cells play a critical role in this process. As part of the innate immune response, TLRs lead to cellular activation and cytokine production with subsequent initiation of an adaptive immune response. TLR7-9 recognize single-stranded RNA, nucleoside analogs and single-stranded CpG-DNA, respectively, and their activation initiates the immune response against viruses and bacteria. Furthermore, the stimulation of these TLRs may be exploited for adjuvant therapy, vaccination and anti-tumor responses. However, a role in the generation or perpetuation of autoimmune diseases such as systemic lupus erythematosus (SLE) has also been suggested. PMID- 18406378 TI - Immune responses induced by spirochetal outer membrane lipoproteins and glycolipids. AB - The class of Spirochetes comprises a wide array of clinically important pathogens, including Treponema pallidum causing syphilis as well as Borrelia burgdorferi, the agent of Lyme disease (LD). Diseases caused by spirochetes are characterized by specific sequelae of host reactions, and also by characteristic antibody response patterns. Over the last decades, research on the interaction of spirochetes with the host's immune system had a strong emphasis on outer membrane lipoproteins. In fact, these structures have been convincingly shown to activate immune cells via CD14 and Toll-like receptor (TLR)-2, and recent data also indicate an interaction with lipopolysaccharide (LPS)-binding protein (LBP). In particular, the interaction of B. burgdorferi with TLR-2 could not only be demonstrated in mice, but was also supported by data showing that genetic variants of TLR-2 in humans influenced the clinical course of LD. However, there is increasing evidence that next to lipoproteins, glycolipids may also play an important role in responses of the immune system towards spirochetes. Diacylglycerol-containing glycolipids exhibiting similarities with lipoteichoic acid (LTA) of Gram-positive bacteria have been demonstrated in various Treponema species, whereas LPS-like glycolipids have been shown to be present in Leptospira. Treponema glycolipids, comparably to lipoproteins and LTA, interact with LBP, CD14 and TLRs. In contrast, complex glycolipids of high molecular weight could not be demonstrated in Borrelia, whereas these bacteria exhibit a number of unique low molecular weight glycolipids. Some of these glycolipids cause strong immediate immune responses, while others appear to be potent antigens for induction of an adaptive immune response. This review summarizes data obtained so far on amphiphilic and hydrophobic molecules from spirochetes regarding structure and influence on innate as well as adaptive immune responses. PMID- 18406379 TI - The zinc finger protein and transcriptional repressor Gfi1 as a regulator of the innate immune response. AB - Gfi1 is a transcriptional repressor with a molecular weight between 47 and 55kDa. The protein has six C-terminal C(2)H(2)-type zinc finger domains and a characteristic stretch of 20 amino acids, called the SNAG-domain, at its N terminus. Expression of Gfi1 ranges from the hematopoietic and lymphoid system to sensory epithelia, lung and parts of the CNS. Gene knockout studies revealed that Gfi1 is essential for the development of granulocytes and plays a role in macrophage-dependent cytokine production, indicating that this protein shares the responsibility for different lines of defense against pathogens. Strikingly, Gfi1 deficient mice are highly sensitive to both endotoxin and bacterial infections and die rapidly after an experimental application of endotoxin or induction of infection with symptoms of septic shock. This sensitivity is mediated by an overproduction of tumor necrosis factor (TNF) and other inflammatory cytokines. The lung could be identified as the principal organ in which the accelerated inflammatory reactions take place in challenged Gfi1-deficient mice. Several lines of experimental evidence support a role of Gfi1 as a regulator of the Toll like receptor (TLR) pathways, and, in general, as an essential modulator preventing an overshooting of the inflammatory response. PMID- 18406380 TI - Immediate lymphotoxin beta receptor-mediated transcriptional response in host defense against L. monocytogenes. AB - The lymphotoxin beta receptor (LTbetaR) mediates crucial signals in host defense against intracellular bacteria and viruses. Mice deficient in LTbetaR readily succumb to infections with Listeria monocytogenes, Mycobacterium tuberculosis and murine cytomegalovirus (mCMV). LTbetaR has been shown to be important for the early induction of interferon (IFN) beta after infection with mCMV. However, up to now, it is not known which host effector molecules are induced in cells of the innate immune system after bacterial infections. In order to address this question, comprehensive transcriptome profiling of LTbetaR-deficient and control splenocytes depleted from T and B lymphocytes was performed and differentially regulated genes were identified. Interestingly, a deficiency in IFNalpha- and IFNgamma-mRNA transcription could be found in LTbetaR-deficient cells leading to a marked failure in the immediate up-regulation of IFN controlled genes. These encompass interferon regulatory factors (IRF1 and IRF7), signal transducer activator of transcription (STAT) proteins (STAT1 and STAT2), chemokines, IFN regulated GTPases (IRGs, GBPs), and IFN-induced protein with tetratricopeptide repeats (IFITs). Thus, the immediate LTbetaR-initiated transcriptional response of innate immune cells carries an IFN signature and is responsible for mounting an effective innate immune response to L. monocytogenes. PMID- 18406381 TI - The IRG proteins: a function in search of a mechanism. AB - The IRG proteins (p47 GTPases) constitute one of the strongest resistance systems known to be active against intracellular pathogens in mice. The proteins are induced by interferons and assemble on phagosomes and parasitophorous vacuoles of a number of different micro-organisms in all cell types assayed. There are presently three experimentally based views as to how they exert their cell autonomous activity against intracellular pathogens: blocking of interferon mediated acceleration of phagosome maturation, induction of autophagic membranes, and direct destruction of the parasitophorous vacuole membrane. Failure of hemopoietic stem cells during infection is associated with targeted deletion of one IRG protein, Irgm1. The significance of this non-cell-autonomous phenotype is discussed. PMID- 18406383 TI - Providing a solid foundation of toxicologic information with many unique and insightful perspectives on their topics. Preface. PMID- 18406382 TI - The innate immune response against Leishmania parasites. AB - Parasites of the genus Leishmania are the causative agents of cutaneous, mucocutaneous or visceral leishmaniasis. The parasite species and host genetic factors determine the quality of the immune response and thereby the outcome of the infection. Here, we summarize previously published and present novel data on several aspects of the early innate immune reaction to Leishmania (L.) major, L. braziliensis and L. infantum, which cause cutaneous, mucocutaneous or visceral leishmaniasis, respectively. We will focus on (1) the effector molecules that contribute to the control of the parasite in the skin, lymph nodes and/or spleen; and (2) on the pattern recognition receptors (Toll-like receptors, TLRs), cell types (myeloid dendritic cells, plasmacytoid dendritic cells), cytokines (IL-12, IFN-alpha/beta), and signaling pathways (Tyk2 kinase) that are necessary for the initial sensing of the parasites and the subsequent development of an efficient NK cell response. PMID- 18406384 TI - Diagnostic sampling and establishing a minimum database in exotic animal toxicology. AB - A toxicologic diagnosis is based on knowledge of the circumstances surrounding a particular case, knowledge of the clinical symptomatology, receipt and evaluation of proper specimens by a qualified laboratory, and judicious interpretation of the laboratory results. Failure to have all necessary ingredients can result in a wrong or missed diagnosis. Many veterinary toxicology laboratories can detect suspected toxicants in feed, tissue, and environmental samples at extremely low concentrations. The ability to detect toxicants at such low levels has often outpaced the ability of the diagnostician to interpret the analytic findings. This article provides guidelines for acquiring a good history, collecting appropriate samples for analysis, and selecting a veterinary analytic laboratory to maximize the probability of making a correct toxicologic diagnosis. PMID- 18406385 TI - Emergency care and managing toxicoses in the exotic animal patient. AB - The poisoned exotic veterinary patient remains a significant challenge to the clinician. A patient presenting with the history of exposure to a toxin or poison should be considered to have a potentially life-threatening problem. This article details the ABCs of emergency medicine including fluid therapy and discusses the principles of neurologic management. The last part of the article provides the clinician with guidelines for decontamination and critical care management of some of the more common toxicoses in exotic animals. PMID- 18406386 TI - Pet bird toxicity and related environmental concerns. AB - Birds may be exposed to toxins through various sources in their everyday environment. Toxicity may occur through inhalation or oral or dermal exposures. Clinicians diagnose and treat these toxicities in an effort to correct the disease of the individual patient. Recognition of toxicity in the avian patient has further significance as it relates to the patient's environment, including the health of other animals, humans, and the ecosystem. While some toxicities, such as lead and zinc toxicosis, are well-documented in avian species, others are limited to anecdotal reports and extrapolation from other species. Continued research is needed in this area of avian medicine to expand our knowledge and improve our ability to diagnose and treat toxic conditions in birds. PMID- 18406387 TI - Raptor toxicology. AB - Birds of prey have demonstrated the negative impact that toxic agents can cause on animal populations and ecosystem dynamics. Lead, cholinesterase inhibitors (eg, organophosphates, carbamates), and anticoagulant rodenticides (eg, brodifacoum) are the most common toxic agents that currently affect the health of wild birds of prey in the United States. For raptors held in captivity, the list of toxic agents expands and includes toxic inhalants such as carbon monoxide and polytetrafluoroethylene. This article provides diagnostic and treatment guidelines for the toxic agents discussed. PMID- 18406388 TI - Waterfowl toxicology: a review. AB - Waterfowl populations may serve as sentinel species for natural and anthropogenic toxicologic problems in the environment. Unfortunately, many toxins cause nonspecific clinical signs, acute mortality, and subtle or no pathologic changes, making toxicologic investigations extremely difficult. The purpose of this article is to review important waterfowl toxins, including heavy metals, pesticides, botulism, mycotoxins, algal toxins, and petroleum oil. When applicable, clinical signs, diagnosis, pathologic findings, and treatment are discussed. Although most of the information in the literature is based on wild waterfowl populations or experimental toxicologic investigations, the information is also applicable to captive waterfowl populations. PMID- 18406389 TI - Toxicology of ferrets. AB - Because of their curious nature and small size, ferrets are at risk for various toxicoses. At present, there is not a great deal of information on specific toxicants in ferrets. This article initially reviews general consideration in treating poisoning in ferrets, such as obtaining history and decontamination. It then discusses some specific agents that appear to be common causes of poisoning in ferrets based on the experience of the ASPCA Animal Poison Control Center. PMID- 18406390 TI - Clinical toxicoses of domestic rabbits. AB - Toxicoses are an uncommon presentation to rabbit practitioners; however, veterinarians who accept rabbits as patients should be familiar with the basic concepts of toxicosis management and the specific syndromes associated with clinical toxicoses. The objective of this article is to present clinically relevant information for veterinarians presented with rabbits exhibiting characteristic signs of toxicosis. In addition, specific mention is made to the most common clinical toxicoses, including lead, chemicals, rodenticides, aflatoxins, and poisonous plants. PMID- 18406391 TI - Poisonings in reptiles. AB - Reptiles are increasingly being kept as pets in American households. The basic principles of emergency medicine are the same for all species, but reptilian species present special diagnostic challenges to veterinary clinicians when they become ill. Reptiles in captivity can become accidentally poisoned in a variety of ways. Veterinarians treating small animal emergencies must make an effort to familiarize themselves with the large body of literature and resources that are developing regarding both nontraditional exotic companion species and advances in toxicology. PMID- 18406392 TI - Toxicology of aquarium fish. AB - Most aquarium fish live in a closed system, so the effects of toxins can be cumulative and devastating. Most cases of toxicity are due to deficiencies in husbandry and tank maintenance. Poor water quality kills more fish than infectious agents, making client education a very important preventive tool for aquatic practitioners. This article includes a discussion of toxicities related to water quality, chemotherapeutics, pesticides, and household substances. PMID- 18406393 TI - Toxic exotics. AB - The purpose of this article is to familiarize the reader with the basic venom components, the pathophysiologic responses of envenomated dogs and cats, and some brief treatment guidelines for envenomations by various exotic "pets." Representative toxic species of reptiles, amphibians, and arthropods are included. The growing trend toward the collection of exotic animals by private owners increases the likelihood that veterinarians will face the challenge of treating an exotic envenomation. PMID- 18406394 TI - Toxicologic information resources for reptile envenomations. AB - The United States is the largest importer of reptiles in the world, with an estimated 1.5 to 2.0 million households keeping one or more reptiles. Snakes account for about 11% of these imports and it has been estimated that as many as 9% of these reptiles are venomous. Envenomations by nonindigenous venomous species are a rare but often serious medical emergency. Bites may occur during the care and handling of legitimate collections found in universities, zoos, or museums. The other predominant source of exotic envenomation is from amateur collectors participating in importation, propagation, and trade of non-native species. This article provides toxicologic information resources for snake envenomations. PMID- 18406395 TI - The interferometric investigations of electromembrane processes. AB - The multi-frequency laser interferometric method for measuring concentration profiles under electrodialysis with ion-exchange membranes including the case of small intermembrane distances and in the presence of ion-conducting spacers is developed. Experimental results obtained by laser interferometric method are in good agreement with the mathematical models of electrodialysis. The amplitude of the concentration field oscillations is shown to increase at the current density exceeding the limiting current density, and these oscillations transfer into the state of dissipative chaos. This instability is presumably a consequence of irregularly distributed heat sources, namely, the Joule heat and the heat of dissociation and recombination of water molecules. PMID- 18406396 TI - Isn't self-assembly a misnomer? Multi-disciplinary arguments in favor of co assembly. AB - Self-assembly is a ubiquitous physicochemical phenomenon. It is inherent to molecular recognition effects in the biological domain, and thus presents a basis for understanding the constitution and dynamic organization of living beings. However, it is argued in this paper that the very notion of self-assembly presents an incorrect and misleading one for the physical effects that it stands for. Self-assembly presents a misnomer as each self-assembly process not only depends on, but also includes a certain degree of ordering in the immediate surrounding of the 'self-assembling' entities. Claims are provided to support the idea that every self-assembly process is, in fact, a co-assembly event, as it implies mutual changes and structuring of both the 'self-assembled' system and its nearest surrounding. This point of view is related to the systemic observation that there could be no physical qualities independent on the context of their existence. In order to illustrate that the concept of co-assembly presents only an instance of a more profound and systemic philosophy of life, the examples of the interplay between internal and external factors of development are provided from the fields of quantum and classical physics, biology, evolutionary theory and psychology. PMID- 18406397 TI - Sodium butyrate stimulates mineralized nodule formation and osteoprotegerin expression by human osteoblasts. AB - OBJECTIVE: Butyric acid (sodium butyrate; BA) is a major metabolic by-product of main periodontopathic bacteria present in subgingival plaque. In the present study, we examined the effects of BA on cell proliferation, alkaline phosphatase (ALPase) activity, mineralized nodule formation, extracellular matrix protein expression, macrophage colony-stimulating factor (M-CSF), and osteoprotegerin (OPG) in normal human osteoblasts. METHODS: The cells were cultured with 0, 10( 8), 10(-6) or 10(-4)M BA for up to 12 days. Mineralized nodule formation was detected by alizarin red staining, and the calcium content in mineralized nodules was determined using a calcium assay kit. The gene and protein expression levels for type I collagen, bone sialoprotein (BSP), osteopontin (OPN), M-CSF, and OPG were examined using real-time PCR and ELISA, respectively. RESULTS: Mineralized nodule formation and the calcium content of mineralized nodules were increased by BA in a dose-dependent manner. Cell proliferation and ALPase activity were not affected by the addition of BA. Following the addition of 10(-4)M BA, the expression levels of BSP, OPN, and OPG increased, whereas the expression levels of type I collagen and M-CSF were not markedly affected. CONCLUSION: These results suggest that BA stimulates bone formation by increasing the production of BSP and OPN, whereas it suppresses osteoclast differentiation by increasing the production of OPG by human osteoblasts. PMID- 18406398 TI - Low plasma brain-derived neurotrophic factor and childhood physical neglect are associated with verbal memory impairment in major depression--a preliminary report. AB - BACKGROUND: Early life stress has been suggested to mediate vulnerability to affective disorders. Animal models of repeated maternal separation have shown reduced brain-derived neurotrophic factor (BDNF) levels in specific brain regions implicated with hypothalamic-pituitary-adrenal axis and memory formation. In addition, BDNF levels are also reduced in major depressive disorder (MDD) and bipolar disorder. The aim of this study was to investigate whether childhood physical neglect (CPN) and plasma BDNF levels would impact on memory performance in adult female subjects with recurrent major depression. METHODS: Recurrent female MDD outpatients with CPN (MDD + CPN, n = 17) and without CPN (MDD, n = 17) and healthy control subjects (n = 15) were assessed for plasma BDNF content and verbal memory performance. Memory was assessed through the logical memory component of the Weschler Memory Scale-Revised for immediate and delayed recall. Brain-derived neurotrophic factor was assessed with enzyme-linked immunosorbent assays (ELISAs). RESULTS: Major depressive disorder patients showed lower plasma BDNF concentrations than healthy control subjects (p < .001). Major depressive disorder + CPN had even lower BDNF levels compared with control subjects and MDD (p < .05). Brain-derived neurotrophic factor levels were negatively related to psychological morbidity and positively correlated to memory performance. Regression models showed that severity of self-reported CPN and low plasma BDNF predicted impairment on immediate verbal recall. Delayed recall impairment was predicted by severity of CPN and depression and memory retention by posttraumatic stress disorder (PTSD) severity symptoms. CONCLUSIONS: Our data suggest that CPN and plasma BDNF are important factors associated with depression and verbal memory performance, particularly with encoding processes. PMID- 18406399 TI - Drug-dependent requirement of hippocampal neurogenesis in a model of depression and of antidepressant reversal. AB - BACKGROUND: Depression and anxiety disorders have been linked to dysfunction of the hypothalamo-pituitary-adrenal (HPA) axis and structural changes within the hippocampus. Unpredictable chronic mild stress (UCMS) can recapitulate these effects in a mouse model, and UCMS-induced changes, including downregulation of hippocampal neurogenesis, can be reversed by antidepressant (AD) treatment. We investigated causality between changes in hippocampal neurogenesis and the effects of both chronic stress and chronic ADs. METHODS: Mice were treated with either a sham procedure or focal hippocampal irradiation to disrupt cell proliferation before being confronted with 5 weeks of UCMS. From the third week onward, we administered monoaminergic ADs (imipramine, fluoxetine), the corticotropin-releasing factor 1 (CRF(1)) antagonist SSR125543, or the vasopressin 1b (V(1b)) antagonist SSR149415 daily. The effects of UCMS regimen, AD treatments, and irradiation were assessed by physical measures (coat state, weight), behavioral testing (Splash test, Novelty-Suppressed feeding test, locomotor activity), and hippocampal BrdU labeling. RESULTS: Our results show that elimination of hippocampal neurogenesis has no effect on animals' sensitivity to UCMS in several behavioral assays, suggesting that reduced neurogenesis is not a cause of stress-related behavioral deficits. Second, we present evidence for both neurogenesis-dependent and -independent mechanisms for the reversal of stress-induced behaviors by AD drugs. Specifically, loss of neurogenesis completely blocked the effects of monoaminergic ADs (imipramine, fluoxetine) but did not prevent most effects of the CRF(1) and the V(1b) antagonists. CONCLUSIONS: Hippocampal neurogenesis might thus be used by the monoaminergic ADs to counteract the effects of stress, whereas similar effects could be achieved by directly targeting the HPA axis and related neuropeptides. PMID- 18406400 TI - Involvement of the nitric oxide-cyclic GMP-protein kinase G-K+ channel pathway in the antihyperalgesic effects of bovine lactoferrin in a model of neuropathic pain. AB - The possible involvement of the nitric oxide (NO)-cyclic GMP (cGMP)-protein kinase G (PKG) pathway on bovine lactoferrin (BLF)-induced spinal antihyperalgesic activity was elucidated in sciatic nerve injured rats. Intrathecal BLF reduced thermal hyperalgesia in a dose-dependent manner. Pretreatment with NG-L-nitro-arginine methyl ester (L-NAME, non-specific inhibitor of NO synthase), 7-nitroindazole (7-NI, neuronal NO synthase inhibitor), 1H-[1,2,4]-oxadiazolo [4,3-a] quinoxalin-1-one (ODQ, guanylyl-cyclase inhibitor), (9S, 10R, 12R)-2,3,9,10,11,12-hexahydro-10-methoxy-2, 9-dimethyl-1 oxo-9, 12-epoxy-1H-diindolo-[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4 i][1,6]benzodiazocine-10-carboxylic acid methyl ester (KT-5823, specific PKG inhibitor) or glybenclamide (ATP-sensitive K+ channel blocker), but not NG-D nitro-arginine methyl ester (D-NAME, an inactive enantiomer of l-NAME), d-Phe-Cys Tyr-d-Trp-Orn-Thr-NH2 (CTOP, selective mu-opioid receptor antagonist) or naloxone (nonselective opioid receptor antagonist) prevented BLF-induced antihyperalgesia. Data suggest that BLF-induced spinal antihyperalgesia could be due to activation of the NO-cGMP-PKG-K+ channel pathway and it is not mediated by mu-opioid receptor in a model of neuropathic pain. PMID- 18406401 TI - Edaravone prevents kainic acid-induced neuronal death. AB - There is growing evidence that free radical generation may play a key role in the neuronal damage induced by prolonged convulsions. Free radical scavengers are known to inhibit neuronal death induced by exposure to excitotoxins. However, this neuroprotective effect has not been demonstrated with treatment after seizures had been stopped. We investigated whether 3-methyl-1-phenyl-2-pyrazolin 5-one, edaravone (Ed), a newly developed free radical scavenger that has been used clinically to treat cerebral infarction, could prevent neuronal loss when administered after the occurrence of seizures in a kainic acid (KA)-induced seizure model. Compared with KA alone, cell loss was significantly reduced when animals received Ed (10 mg/kg i.v.) just after seizures, and when Ed was administered both 60 min before (30 mg/kg i.p.) and after KA injection. Combined before-and-after treatment with Ed significantly ameliorated the KA-induced decrease of glutathione and blocked the KA-induced increase of 4-hydroxy-2 nonenal (HNE). Because before-and-after treatment with Ed significantly lessened the KA-induced increase of HNE, Ed may exert its neuroprotective effect by inhibiting lipid peroxidation. However, post-treatment with Ed prevented neuronal cell loss, while HNE and glutathione levels did not differ from those in animals without Ed, so a mechanism other than free radical scavenging must be involved in the prevention of cell loss. Patients who develop status epilepticus are unlikely to receive adequate antioxidant therapy before the onset, so it is an advantage that Ed can prevent neuronal death even when administered after seizures. PMID- 18406402 TI - Stereoselective glycosylations using benzoylated glucosyl halides with inexpensive promoters. AB - Reactions of O-benzoylated glucopyranosyl halide (I, Br), isolated or generated in situ from per-benzoylated glucose (8a) and trimethylsilyl halide, with various alcohols were efficiently promoted by zinc halide (Cl, Br) or N-bromosuccinimide with a catalytic ZnI(2) to give the corresponding 1,2-trans-beta-glucosides in good to high yields. When the anomeric halogenation of 8a was carried out in the presence of reactive alcohols, 1,2-cis-alpha-glucosides were selectively formed. PMID- 18406403 TI - Connexin43 (GJA1) is required in the population of dividing cells during fin regeneration. AB - In zebrafish, mutations in the gap junction gene connexin43 lead to short bony fin ray segments that give rise to the short fin phenotype. The sof(b123) mutant exhibits fins that are half the length of wild-type fins and have reduced levels of cx43 mRNA. We find that sof(b123) regenerating fins exhibit reduced levels of cell proliferation. Interestingly, the number of dividing cells per unit length of fin growth is similar between wild-type and mutant fins, suggesting that the number of cells that enter the cell cycle is specifically affected in sof(b123). Expression of cx43 is identified in mitotic cells, which further suggests that Cx43 may contribute to establishing or maintaining the population of dividing cells. Indeed, missense alleles exhibiting high or low levels of gap junctional communication reveal a correlation between defects in direct cell-cell communication, cell proliferation, and segment length. Finally, targeted gene knockdown of cx43 in adult regenerating fins recapitulates the sof(b123) phenotype, revealing that the loss of Cx43 is sufficient to reduce both cell proliferation and segment length. We hypothesize that the level of gap junctional intercellular communication among dividing cells regulates the level of cell proliferation and ultimately regulates bone growth. PMID- 18406404 TI - Lengsin expression and function during zebrafish lens formation. AB - A zebrafish ortholog of human lengsin was identified by EST analysis of an adult lens cDNA library. During zebrafish development, lengsin transcription is first detected at 24 h post-fertilization (hpf). Immunolocalization, using polyclonal antiserum generated against a Lengsin bacterial fusion protein, detects lens specific protein in whole-mount embryos at 30 hpf. Lengsin expression in zebrafish follows the temporal expression of the alphaA- alphaB1- and betaB1 crystallin proteins in the lens. At 72 hpf, Lengsin is localized to a subpopulation of differentiating secondary fiber cells, while no expression is detected in the lens epithelial cells or central lens fibers. In the adult lens, Lengsin is restricted to a narrow band of cortical fibers and co-localizes with actin at the lateral faces of these interdigitating cells. Stable transgenic lines, using a 3 kb lengsin genomic fragment to regulate EGFP expression, recapitulate the Lengsin temporal and spatial expression patterns. Lengsin function in zebrafish lens formation was examined by antisense morpholino mediated translation and mRNA splice inhibition. At 72 hpf, the lengsin morphant lenses are reduced in size and exhibit separations within the cortex due to defects in secondary fiber morphogenesis. The location of the morphant lens defects correlates with the Lengsin protein localization at this age. These results demonstrate Lengsin is required for proper fiber cell differentiation by playing roles in either cell elongation or the establishment of cell interactions. PMID- 18406405 TI - Transforming growth factor-beta induces cellular injury in experimental diabetic neuropathy. AB - The mechanism/s leading to diabetic neuropathy are complex. Transforming growth factor-beta1 (TGF-beta1) has been associated with diabetic nephropathy and retinopathy but not neuropathy. In this study, changes in TGF-beta isoforms were examined in vivo and in vitro. Two groups of animals, streptozotocin diabetic with neuropathy and non-diabetic controls were examined at 4 weeks (n=10/group) and 12 weeks (n=8/group). In diabetic DRG using quantitative real-time PCR (QRT PCR), TGF-beta1 and TGF-beta2 mRNA, but not TGF-beta3, was increased at 4 and 12 weeks. In sciatic nerve TGF-beta3 mRNA was primarily increased. Immunohistochemistry (DRG) and immunoblotting (sciatic nerve) showed similar differential protein expression. In sciatic nerve TGF-beta formed homo- and hetero-dimers, of which beta(2)/beta(3), beta(1)/beta(1), and beta(1)/beta(3) were significantly increased, while that of the TGF-beta(2)/beta(2) homodimer was decreased, in diabetic compared to non-diabetic rats. In vitro, pretreatment of embryonic DRG with TGF-beta neutralizing antibody prevents the increase in total TGF-beta protein observed with high glucose using immunoblotting. In high glucose conditions, combination with TGF-beta2>beta1 increases the percent of cleaved caspase-3 compared to high glucose alone and TGF-beta neutralizing antibody inhibits this increase. Furthermore, consistent with the findings in diabetic DRG and nerve, TGF-beta isoforms applied directly in vitro reduce neurite outgrowth, and this effect is partially reversed by TGF-beta neutralizing antibody. These findings implicate upregulation of TGF-beta in experimental diabetic peripheral neuropathy and indicate a novel mechanism of cellular injury related to elevated glucose levels. In combination, these findings indicate a potential new target for treatment of diabetic peripheral neuropathy. PMID- 18406406 TI - Farnesyl diphosphate synthase localizes to the cytoplasm of Trypanosoma cruzi and T. brucei. AB - The farnesyl diphosphate synthase (FPPS) has previously been characterized in trypanosomes as an essential enzyme for their survival and as the target for bisphosphonates, drugs that are effective both in vitro and in vivo against these parasites. Enzymes from the isoprenoid pathway have been assigned to different compartments in eukaryotes, including trypanosomatids. We here report that FPPS localizes to the cytoplasm of both Trypanosoma cruzi and T. brucei, and is not present in other organelles such as the mitochondria and glycosomes. PMID- 18406407 TI - Gossypiboma of the urinary bladder. PMID- 18406408 TI - The cost of caring: commentary on Corbin (2008). PMID- 18406410 TI - Failure of mineralized collagen fibrils: modeling the role of collagen cross linking. AB - Experimental evidence demonstrates that collagen cross-linking in bone tissue significantly influences its deformation and failure behavior yet difficulties exist in determining the independent biomechanical effects of collagen cross linking using in vitro and in vivo experiments. The aim of this study is to use a nano-scale composite material model of mineral and collagen to determine the independent roles of enzymatic and non-enzymatic cross-linking on the mechanical behavior of a mineralized collagen fibril. Stress-strain curves were obtained under tensile loading conditions without any collagen cross-links, with only enzymatic cross-links (modeled by cross-linking the end terminal position of each collagen domain), or with only non-enzymatic cross-links (modeled by random placement of cross-links within the collagen-collagen interfaces). Our results show enzymatic collagen cross-links have minimal effect on the predicted stress strain curve and produce a ductile material that fails through debonding of the mineral-collagen interface. Conversely, non-enzymatic cross-links significantly alter the predicted stress-strain response by inhibiting collagen sliding. This inhibition leads to greater load transfer to the mineral, which minimally affects the predicted stress, increases modulus and decreases post-yield strain and toughness. As a consequence the toughness of bone that has more non-enzymatically mediated collagen cross-links will be drastically reduced. PMID- 18406409 TI - In vitro evaluations of antimalarial drugs and their relevance to clinical outcomes. AB - Plasmodium falciparum resistance to the former first-line antimalarials chloroquine and sulfadoxine/pyrimethamine has reached critically high levels in many malaria-endemic regions. This has spurred the introduction of several new artemisinin-based combination therapies (ACTs) that display excellent potency in treating drug-resistant malaria. Monitoring for the emergence of drug resistant P. falciparum is important for maximising the clinically effective lifespan of ACTs. Here, we provide a commentary on the article by Kaddouri et al., published in this issue of the International Journal of Parasitology, which documents the levels of susceptibility to ACT drugs and chloroquine in P. falciparum isolates from Mali. These authors report that some isolates approached a proposed in vitro threshold of resistance to monodesethyl-amodiaquine (the principal effective metabolite of amodiaquine, an important ACT partner drug), and establish baseline levels of susceptibility to the ACT drugs dihydroartemisinin and lumefantrine. The majority of clinical isolates manifested in vitro resistance to chloroquine. The authors also show good concordance between field-based assays employing a non radioactive lactate dehydrogenase-based method of determining in vitro drug IC(50) values and the well-established [(3)H]hypoxanthine-based radioactive method. This work illustrates a good example of drug resistance surveillance, whose global coordination is being championed by the World Antimalarial Resistance Network. Our current opinion also more generally discusses the complexities inherent to conducting in vitro investigations with P. falciparum patient isolates and correlating these findings with treatment outcome data. PMID- 18406411 TI - Development and validation of a method for quantitative determination of econazole nitrate in cream formulation by capillary zone electrophoresis. AB - A simple, fast, inexpensive and reliable capillary zone electrophoresis (CZE) method for the determination of econazole nitrate in cream formulations has been developed and validated. Optimum conditions comprised a pH 2.5 phosphate buffer at 20 mmol L(-1) concentration, +30 kV applied voltage in a 31.5 cm x 50 microm I.D. capillary. Direct UV detection at 200 nm led to an adequate sensitivity without interference from sample excipients. A single extraction step of the cream sample in hydrochloric acid was performed prior to injection. Imidazole (100 microg mL(-1)) was used as internal standard. Econazole nitrate migrates in approximately 1.2 min. The analytical curve presented a coefficient of correlation of 0.9995. Detection and quantitation limits were 1.85 and 5.62 microg mL(-1), respectively. Excellent accuracy and precision were obtained. Recoveries varied from 98.1 to 102.5% and intra- and inter-day precisions, calculated as relative standard deviation (RSD), were better than 2.0%. The proposed CZE method presented advantageous performance characteristics and it can be considered suitable for the quality control of econazole nitrate cream formulations. PMID- 18406412 TI - High-speed, low-pressure gas chromatography-mass spectrometry for essential oil analysis. AB - Analysis of parsley and fennel essential oils was performed by using low-pressure gas chromatography-mass spectrometry (GC-MS). The low-pressure instrument configuration was achieved by fitting a GC-MS instrument with a 530microm I.D. capillary column and an appropriate capillary restrictor at the inlet of the column. Comparison of the performance of the low-pressure GC-MS setup was made with fast GC-MS using a narrow-bore capillary column. By comparing the two approaches side-by-side the benefits of low-pressure GC-MS for characterisation of moderately complex essential oils comprising less than 50 detectable components can be fully appreciated. Although efficiency is sacrificed, the improved sample capacity of the 530microm I.D. column leads to higher peak intensities and in-turn better mass spectral library matching thus providing highly satisfactory results. PMID- 18406413 TI - Extraction of metal ions in aqueous polyethylene glycol-inorganic salt two-phase systems in the presence of inorganic extractants: correlation between extraction behaviour and stability constants of extracted species. AB - The use of aqueous polyethylene glycol-inorganic salt two-phase systems for the extraction of metal ions has a great potential due to their durability, non toxicity and relative low cost. The aqueous phases can be easily separated by centrifugation, and the operation is possible in a range of experimental conditions. The experimental results have shown that for a given aqueous two phase system, the extraction behaviour of metal ions in presence of inorganic extractants is mainly dependent on the stability of extracted species. In this paper we review our results obtained at metal ion extraction using inorganic extractants and discuss three major types of extraction behaviours. PMID- 18406414 TI - Multifactorial optimization approach for the determination of polycyclic aromatic hydrocarbons in river sediments by gas chromatography-quadrupole ion trap selected ion storage mass spectrometry. AB - A procedure for the determination of very low polycyclic aromatic hydrocarbons (PAHs) concentrations in sediment samples has been developed by gas chromatography-quadrupole ion trap mass spectrometry (GC-QIT MS) after extraction with dichloromethane and purification by using silica gel cleanup. Identification and quantification of analytes were based on the selected ion storage (SIS) strategy using deuterated PAHs as internal standards. In order to search out the main factors affecting the SIS mass spectrometry efficiency, four MS parameters, including target total ion count (TTIC), waveform amplitude (WA), transfer line (XLT) and ion trap temperatures (ITT) were subjected to a complete multifactorial design. The most relevant parameters obtained (TTIC and WA) were optimized by a rotatable and orthogonal composite design. Optimum values for these parameters were selected for the development of the method involving PAH determination in sediment samples. The optimized method exhibited a range of 111-760% higher signal-to-noise (S/N) ratios for PAHs in comparison with the method operated by the default conditions, demonstrating that the multifactorial optimization contributed to substantially improve the sensitivity of the GC-QIT MS determination. The accuracy of the method was verified by analyzing NWRI EC-3 certified reference material (Lake Ontario sediment). The selectivity, sensitivity (limits of quantification were in the range of 0.02-11.0 ng g(-1)), accuracy (recoveries >or=77%) and precision (RSD25 ng/m3 Hg0 in 14% of buildings in study areas compared to only one reference building. In the latter we identified an accidental mercury spill from a bottle that had been brought into the building. Both the mean and maximum indoor mercury vapor levels were greater in the study communities than in the reference community. In all communities, we observed mean indoor Hg0 vapor concentration greater than outdoors, although in two-thirds of buildings, indoor levels did not exceed the area-specific outdoor upper-limit concentration. CONCLUSION: After controlling for factors that might influence Hg0 vapor levels, the most plausible explanation for greater Hg0 levels in the study area is a relationship to cultural use of mercury. None of the measured levels exceeded the ATSDR minimum risk level for residences of 200 ng/m3 Hg0 although levels in living quarters might be greater than those in the common areas. PMID- 18406446 TI - Functional microbial diversity of the railway track bed. AB - Railways constitute relatively unexplored microbial habitats. Little is known about the amounts, activities or distributions of microorganisms and their associated heterotrophic capabilities on railway embankments. The aim of this study was to investigate the microbiology of two Swedish railway tracks in order to fill some of the gaps in the available information. We estimated microbial biomass by means of substrate-induced respiration, microbial activity as basal respiration (BR) and as a kinetically derived parameter (r) hypothesised to correspond to the active fraction of the microbial biomass. It was confirmed that the microbial biomass and activity were low as compared with agricultural soils and that their distributions were distinctly positively skewed. Spatial Kriging revealed that covariance structures were sustained on a scale smaller than the employed sampling grid (<1 m). Substrate richness (SR), as measured with Biolog ECO plates, was used as a quantitative measure of functional diversity. SR correlated to microbial activity and SIR, indicating that functional traits were lacking where the microbial biomass was low or less active. The dependence of microbial activities on basic soil characteristics were inferred by separately designed general linear models. Water content was found to be the most important factor moderating basal respiration and functional diversity, whereas the organic matter content was identified as the most important covariate for SIR. Multivariate analysis of the carbon source utilisation patterns of the Biolog plates with equivalent average well-colour development revealed homogenous substrate utilisation among samples. This indicates that the microbial functional potential is randomly distributed in the railway track bed. In combination, our findings imply that the ecosystem functionality of railway embankments may be seriously hampered as compared with agricultural soils. This has consequences for the risk assessment of herbicides applied to railways. PMID- 18406447 TI - Changes in arsenic speciation through a contaminated soil profile: a XAS based study. AB - An impacted soil located near an industrial waste site in the Massif Central near Auzon, France, where arsenical pesticides were manufactured, has been studied in order to determine the speciation (chemical forms) of arsenic as a function of soil depth. Bulk As concentrations range from 8780 mg kg(-1) in the topsoil horizon to 150 mg kg(-1) at 60 cm depth. As ores (orpiment As2S3, realgar AsS, arsenopyrite FeAsS) and former Pb- and Al-arsenate pesticides have been identified by XRD at the site and are suspected to be the sources of As contamination for this soil. As speciation was found to vary with depth, based on XRD, SEM-EDS, EPMA measurements and selective chemical extractions. Based on oxalate extraction, As is mainly associated with amorphous Fe oxides through the soil profile, except in the topsoil horizons where As is hosted by another phase. SEM-EDS and EPMA analyses led to the identification of arseniosiderite (Ca2Fe3+3(AsVO4)3O2.3H2O), a secondary mineral that forms upon oxidation of primary As-bearing minerals like arsenopyrite, in these topsoil horizons. These mineralogical and chemical results were confirmed by synchrotron-based X-ray absorption spectroscopy. XANES spectra of soil samples indicate that As occurs exclusively as As(V), and EXAFS results yield direct evidence of changes in As speciation with depth. Linear combination fits of EXAFS spectra of soil samples with those of various model compounds indicate that As occurs mainly As-bearing Fe(III)-(hydr)oxides (65%) and arseniosiderite (35%) in the topsoil horizon (0-5 cm depth). Similar analyses also revealed that there is very little arseniosiderite below 15 cm depth and that As(V) is associated primarily with amorphous Fe oxides below this depth. This vertical change of As speciation likely reflects a series of chemical reactions downward in the soil profile. Arseniosiderite, formed most likely by oxidation of arsenopyrite, is progressively dissolved and replaced by less soluble As-bearing poorly ordered Fe oxides, which are the main hosts for As in well aerated soils. PMID- 18406448 TI - Breast adenocarcinoma MCF-7 cell line induces spontaneous osteoclastogenesis via a RANK-ligand-dependent pathway. AB - The metastasis of breast cancer to the skeleton is a serious clinical problem resulting in hypercalcemia, bone fragility and insurmountable pain. The invasion of bony tissue by neoplastic cells usually very rapidly affects the balance between bone apposition and bone resorption. In order to elucidate a mechanism for cancer-induced osteoclastogenesis, cells from a human breast cancer line, MCF 7, were directly co-cultured with murine monocytes RAW 264.7 type CRL 2278. Compared with controls, co-culture of MCF-7 induced differentiation of multinucleated cells by membrane-bound and soluble receptor activator of NF-kB ligand (RANKL) as quantified by ELISA, Western blot analysis, transmission electron microscopy (TEM), and immunocytochemistry. The aim of this study was to determine an in vitro model system of MCF-7 human breast cancer cells grown together with monocytes to show that expression of RANKL promotes osteoclastogenesis, which may indicate a mechanism for the development of osteolytic lesions in breast cancer bone metastasis. PMID- 18406444 TI - DNA mismatch repair: molecular mechanism, cancer, and ageing. AB - DNA mismatch repair (MMR) proteins are ubiquitous players in a diverse array of important cellular functions. In its role in post-replication repair, MMR safeguards the genome correcting base mispairs arising as a result of replication errors. Loss of MMR results in greatly increased rates of spontaneous mutation in organisms ranging from bacteria to humans. Mutations in MMR genes cause hereditary nonpolyposis colorectal cancer, and loss of MMR is associated with a significant fraction of sporadic cancers. Given its prominence in mutation avoidance and its ability to target a range of DNA lesions, MMR has been under investigation in studies of ageing mechanisms. This review summarizes what is known about the molecular details of the MMR pathway and the role of MMR proteins in cancer susceptibility and ageing. PMID- 18406449 TI - An immunohistochemical study of the endocrine pancreas of the African ice rat, Otomys sloggetti robertsi. AB - The African ice rat, Otomys sloggetti robertsi, is a member of the subfamily Otomyinae, in the superfamily of Muroidea, to which all rodents belong. Very little is known about this unique family of rodents. The study reported here examines the endocrine pancreas of this species using immunohistochemical techniques. The islets of Langerhans were scattered in the exocrine pancreas and tended to be quite small. Scattered single endocrine cells (mostly immunoreactive for insulin) were found in the exocrine pancreas and were not generally associated with ducts (as marked by pan-cytokeratin labeling). The normal islet architecture of insulin in the center and glucagon, somatostatin (SS) and pancreatic polypeptide (PP) in the rim was observed, but the islets tended to have 2-3 layers of glucagon immunoreactive cells. Examining for rarer endocrine cell types, we found that cocaine amphetamine regulated transcript (CART) immunoreactive cells were co-localized with SS; and peptide YY (PYY) immunoreactive cells could be found that were singly immunoreactive or co localized with either PP or glucagon. Ghrelin cells were not found. MafA co localized only with the insulin cells, while MafB, which localizes to the glucagon cells, also showed a low level of immunoreactivity in most insulin immunoreactive cells. The Nkx family of transcription factors (Nkx6.1 and 2.2) and PDX-1 were all detected in the pancreas in a similar manner to that seen in mouse and rat. In conclusion, the endocrine pancreas of the African ice rat is quite similar to that of other studied rodents, but these animals have more glucagon and SS cells than rat (Rattus) or mouse (Mus) species. PMID- 18406450 TI - A cytoarchitectonic study of the brain of a perciform species, the sea bass (Dicentrarchus labrax): the midbrain and hindbrain. AB - This study is the third part of a comprehensive series of publications on the cytoarchitectonic organization of the brain of the European sea bass, Dicentrarchus labrax. This study provides an atlas of the brain stem based on Nissl-stained transverse sections as well as a description of cell masses and a discussion on comparative aspects of brain stem nuclei, including methodological studies in other species. By external examination, the sea bass exhibits a prominent Optic tectum and Corpus cerebelli as expected in a predator species with a highly developed visual system. However, no hypertrophy of the facial and vagal lobes was observed as reported in other non-perciform teleosts. The general organization pattern of the mesencephalon and rhombencephalon of the sea bass brain resembles that reported for other perciform teleosts. However, the Valvula cerebelli has been subdivided into anterior, central and posterior parts. In addition, the ventricular surface of the granular layer of the Valvula cerebelli appears to be in contact with those of the Torus longitudinalis. This cell apposition could be interpreted as a direct connection, but more studies demonstrating the absence of ependyma between both structures are needed. Furthermore, we have tentatively described the electro/mechano receptive pre eminential nucleus in the rhombencephalon of the sea bass. This study completes one of the few descriptions, as well as the most complete and detailed available, of the brain of any marine perciform species. PMID- 18406451 TI - Why a clinician may be interested and involved in computational medicine? PMID- 18406452 TI - Life and mechanosensitivity. PMID- 18406453 TI - Experimental and computational studies of strain-conduction velocity relationships in cardiac tissue. AB - Velocity of electrical conduction in cardiac tissue is a function of mechanical strain. Although strain-modulated velocity is a well established finding in experimental cardiology, its underlying mechanisms are not well understood. In this work, we summarized potential factors contributing to strain-velocity relationships and reviewed related experimental and computational studies. We presented results from our experimental studies on rabbit papillary muscle, which supported a biphasic relationship of strain and velocity under uni-axial straining conditions. In the low strain range, the strain-velocity relationship was positive. Conduction velocity peaked with 0.59 m/s at 100% strain corresponding to maximal force development. In the high strain range, the relationship was negative. Conduction was reversibly blocked at 118+/-1.8% strain. Reversible block occurred also in the presence of streptomycin. Furthermore, our studies revealed a moderate hysteresis of conduction velocity, which was reduced by streptomycin. We reconstructed several features of the strain-velocity relationship in a computational study with a myocyte strand. The modeling included strain-modulation of intracellular conductivity and stretch activated cation non-selective ion channels. The computational study supported our hypotheses, that the positive strain-velocity relationship at low strain is caused by strain-modulation of intracellular conductivity and the negative relationship at high strain results from activity of stretch-activated channels. Conduction block was not reconstructed in our computational studies. We concluded this work by sketching a hypothesis for strain-modulation of conduction and conduction block in papillary muscle. We suggest that this hypothesis can also explain uni-axially measured strain-conduction velocity relationships in other types of cardiac tissue, but apparently necessitates adjustments to reconstruct pressure or volume related changes of velocity in atria and ventricles. PMID- 18406454 TI - Drugs that interact with cardiac electro-mechanics: old and new targets for treatment. AB - The concept of mechano-electrical feedback was derived from the observation that a short stretch applied to the beating heart can invoke an electrical response in the form of an afterdepolarization or a premature ventricular beat. More recent work has identified stretch-activated channels whose specific inhibition might help to treat atrial fibrillation in the near future. But the interaction between electrical and mechanical function of the heart is a continuum from short-term (within milliseconds) to long-term (within weeks or months) effects. The long term effects of pressure overload have been well-described on the molecular and cellular level, and substances that interact with these processes are used in clinical routine in the care of patients with cardiac hypertrophy and heart failure. These treatments help to prevent lethal arrhythmias (sudden death) and potentially atrial fibrillation. The intermediate interaction between mechanical and electrical function of the heart is less well-understood. Several recently identified regulatory mechanisms may provide novel antiarrhythmic targets associated with the "intermediate" response of the myocardium to stretch. PMID- 18406456 TI - Linking Global Youth Tobacco Survey (GYTS) data to the WHO Framework Convention on Tobacco Control (FCTC): the case for Mexico. AB - BACKGROUND: The adoption of the World Health Organization Framework Convention on Tobacco Control (WHO FCTC) in 2004 marked a critical achievement in efforts to stem the tobacco epidemic in Mexico. The Global Youth Tobacco Survey (GYTS) findings are useful for evaluating achievement of FCTC articles and designing tobacco control programs. OBJECTIVE: To use data from the GYTS conducted in 21 Mexican cities between 2000 and 2005 to monitor Articles in the WHO FCTC. METHODS: The GYTS uses a two-stage cluster sample survey design that produces representative samples of students aged 13-15 years enrolled in public, private and technical schools. The survey was undertaken at 542 schools in 21 cities. The GYTS surveyed 43,950 students during 2000-2005. RESULTS: The current smoking rate ranged from 10.7% to 29.4%. Among never smokers, susceptibility to initiate smoking ranged from 20.2% to 34.4%. Among current smokers, the percentage who bought their cigarettes in a store was above 40% in 6 cities, but significantly declined over five years in the only city with two assessments (Monterrey). Exposure to secondhand smoke in public places was greater than 50% in 15 of the 21 cities. Over 80% of students in all 21 cities reported that they saw of advertisements for cigarettes on billboards. CONCLUSION: Using determinants measured by GYTS in Mexico, the government can monitor the impact of enforcing various provisions of the National Health Law and the progress made in achieving the goals of the WHO FCTC and the Regional strategy. When these goals are met, tobacco consumption and exposure in Mexico will have declined substantially. PMID- 18406455 TI - Tensegrity-based mechanosensing from macro to micro. AB - This article is a summary of a lecture on cellular mechanotransduction that was presented at a symposium on "Cardiac Mechano-Electric Feedback and Arrhythmias" that convened at Oxford, England in April 2007. Although critical mechanosensitive molecules and cellular components, such as integrins, stretch activated ion channels, and cytoskeletal filaments, have been shown to contribute to the response by which cells convert mechanical signals into a biochemical response, little is known about how they function in the structural context of living cells, tissues and organs to produce orchestrated changes in cell behavior in response to stress. Here, studies are reviewed that suggest our bodies use structural hierarchies (systems within systems) composed of interconnected extracellular matrix and cytoskeletal networks that span from the macroscale to the nanoscale to focus stresses on specific mechanotransducer molecules. A key feature of these networks is that they are in a state of isometric tension (i.e., experience a tensile prestress), which ensures that various molecular-scale mechanochemical transduction mechanisms proceed simultaneously and produce a concerted response. These features of living architecture are the same principles that govern tensegrity (tensional integrity) architecture, and mathematical models based on tensegrity are beginning to provide new and useful descriptions of living materials, including mammalian cells. This article reviews how the use of tensegrity at multiple size scales in our bodies guides mechanical force transfer from the macro to the micro, as well as how it facilitates conversion of mechanical signals into changes in ion flux, molecular binding kinetics, signal transduction, gene transcription, cell fate switching and developmental patterning. PMID- 18406457 TI - Evaluation of three-dimensional scaffolds prepared from poly(3-hydroxybutyrate-co 3-hydroxyhexanoate) for growth of allogeneic chondrocytes for cartilage repair in rabbits. AB - Articular cartilage repair using tissue engineering approach generally requires the use of an appropriate scaffold architecture that can support the formation of cartilage tissue. In this investigation, the potential of three-dimensional scaffolds made of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) was evaluated in rabbit articular cartilage defect model. Engineered PHBHHx cartilage constructs inoculated in vitro with rabbit chondrocytes for 30 days were examined. Subsequently the constructs inoculated with chondrocytes for 10 days were selected for transplantation into rabbits. After 16 weeks of in vivo implantation, both the engineered cartilage constructs and the bare scaffolds were found to be filled the defects with white cartilaginous tissue, with the engineered constructs showing histologically good subchondral bone connection and better surrounding cartilage infusion. Owing to pre-seeded chondrocytes in the PHBHHx scaffolds, better surface integrality and more accumulation of extracellular matrix (ECM) including type II collagen and sGAG were achieved in the engineered cartilage constructs. The repaired tissues possessed an average compressive modulus of 1.58MPa. For comparison, the defects without repair treatments still showed defects with fibrous tissues. These results demonstrated that PHBHHx is a useful material for cartilage tissue engineering. PMID- 18406458 TI - The effect of integrin-specific bioactive coatings on tissue healing and implant osseointegration. AB - Implant osseointegration, defined as bone apposition and functional fixation, is a requisite for clinical success in orthopaedic and dental applications, many of which are restricted by implant loosening. Modification of implants to present bioactive motifs such as the RGD cell-adhesive sequence from fibronectin (FN) represents a promising approach in regenerative medicine. However, these biomimetic strategies have yielded only marginal enhancements in tissue healing in vivo. In this study, clinical-grade titanium implants were grafted with a non fouling oligo(ethylene glycol)-substituted polymer coating functionalized with controlled densities of ligands of varying specificity for target integrin receptors. Biomaterials presenting the alpha5beta1-integrin-specific FN fragment FNIII 7-10 enhanced osteoblastic differentiation in bone marrow stromal cells compared to unmodified titanium and RGD-presenting surfaces. Importantly, FNIII 7 10-functionalized titanium significantly improved functional implant osseointegration compared to RGD-functionalized and unmodified titanium in vivo. This study demonstrates that bioactive coatings that promote integrin binding specificity regulate marrow-derived progenitor osteoblastic differentiation and enhance healing responses and functional integration of biomedical implants. This work identifies an innovative strategy for the rational design of biomaterials for regenerative medicine. PMID- 18406459 TI - pPSY: a vector for the stable cloning and expression of streptomycete single gene phenotypes in Escherichia coli. AB - pPSY is a 12kb cloning vector derived from the IncW plasmid R388, which provides a rapid and easy way to stably clone phenotypes encoded in DNA segments <10kb. In the present study three different genes were amplified by PCR, cloned into pGEM-T Easy and sub-cloned into the EcoRI site of pPSY. The first gene, vioA, is a FAD dependent l-tryptophan amino acid oxygenase from the high G+C Gram-negative bacterium Chromobacterium violaceum. VioA is involved in the synthesis of the indolocarbazole antitumour antibiotic violacein. It was found that vioA was strongly expressed in Escherichia coli from its native promoter. Two other genes encoding recombinase A (recA) and an amylase (amyA), derived from the high G+C Gram-positive streptomycete, Streptomyces lividans, were also tested. Despite recA lacking its native promoter sequence, it was strongly expressed in E. coli using the lac promoter of pGEM-T Easy. Similar to vioA, S. lividansamyA was strongly expressed in E. coli from its native promoter. Unlike pGEM-T Easy, pPSY stably maintained all three genes without the requirement for antibiotic selection. These results demonstrate the applicability of pPSY as a stable amplicon cloning vector for the expression of heterologous genes in E. coli. PMID- 18406461 TI - Comments to the article by Ergene et al. "Genotoxic biomonitoring study of population residing in pesticide contaminated regions in Goksu Delta: micronucleus, chromosomal aberrations and sister chromatid exchanges". PMID- 18406460 TI - Application of a microbiotests battery for complete toxicity assessment of rivers. AB - Acute hazard classification based on selected microbiotests was proposed to assess and compare the toxicity of rivers including surface-water, sediment and soil from floodplains. No direct relationship between the classification of pollution for surface-water based on physical-chemical parameters and proposed acute hazard classification based on organisms' sensitivity was observed. The quality of water according to hazard classification was better than in the mandatory classification of pollution, with the domination of Class II (slight acute hazard). The samples of sediment and soil were more toxic and represented Class II or Class III (acute hazard). The results indicated a need to complete the mandatory monitoring of surface-water in rivers with biological monitoring with toxicity assessment of rivers including water, sediment and soil from floodplains based on acute hazard classification. This integral approach enables a complete evaluation of the toxicity of aquatic life together with an estimation of negative changes in river systems. PMID- 18406462 TI - Sources of hydrocarbons in sediments of the Mandovi estuary and the Marmugoa harbour, west coast of India. AB - Surface sediments were collected from various locations of the Mandovi estuary and the Marmugoa harbour. Sediments were analysed for organic carbon (OC), total lipids, n-alkanes concentration and composition. Concentrations of OC, total lipids and n-alkanes varied spatially and ranged from 1 to 2.5%, 176 to 1413 microg/g dry weight (dw) sediments, and 0.8 to 3.2 microg/g dw sediments of the Mandovi estuary, respectively; and from 0.6 to 2.9%, 233 to 1448 microg/g dw sediments, and 1.6 to 10.7 microg/g dw sediments in the Marmugoa harbour, respectively. Long chain, odd carbon n-alkanes (C23-C33) in the Mandovi estuary, whereas short chain, even carbon n-alkanes (C11-C21) in the Marmugoa harbour sediments were more abundant. The total HC concentrations, n-alkane composition, CPI, UCM and other evaluation indices suggest the dominance of terrestrial hydrocarbons in the estuarine while petroleum derived hydrocarbons in the harbour sediments. This conclusion was further supported by the abundance of hopanes with C29 to C34 alpha, beta compounds and steranes with C27, C28 and C29 compounds in the harbour sediments. PMID- 18406463 TI - Native, amyloid fibrils and beta-oligomers of the C-terminal domain of human prion protein display differential activation of complement and bind C1q, factor H and C4b-binding protein directly. AB - Prion protein (PrP) is an endogenous protein involved in the pathogenesis of bovine spongiform encephalopathy and Creutzfeldt-Jakob disease. Murine PrP has been reported to bind C1q and activate the classical pathway of complement in a copper-dependent manner. Here we show that various conformational isoforms (native, amyloid fibrils, and beta-oligomers) of recombinant human PrP (90-231 and 121-231) bind C1q and activate complement. PrP binds both the globular head and collagenous stalk domains of C1q. Native, beta-oligomeric and amyloid fibrils of PrP all activate the classical and alternative pathways of complement to different extent. However, they do not trigger the lectin pathway. Of the tested PrP conformational isoforms we find that beta-oligomers bind C1q and activate complement most strongly. Membrane attack complex formation initiated by PrP is subdued in comparison to deposition of early complement components. This is most likely attributed to the interaction between human PrP and complement inhibitors factor H and C4b-binding protein. Accordingly, PrP-triggered complement activation in the terminal pathway was increased in serum lacking C4b-binding protein. Taken together the present study indicates that complement activation may be an important factor in human prion diseases, suggesting that complement induced activities may prove relevant therapeutic targets. PMID- 18406464 TI - Leishmania initially activates but subsequently down-regulates intracellular mitogen-activated protein kinases and nuclear factor-kappaB signaling in macrophages. AB - The complex interactions between Leishmania and macrophages are central to the outcome of parasite infection. Disrupting signaling molecules to impair macrophage function, is a subversive strategy used by several pathogens. In the present study, we show that the initial contact of Leishmania with human naives macrophages and murine Raw264.7 macrophage cell line induced a rapid and transient activation of extracellular-signal-regulated kinases 1 and 2 (ERK1/2) and p38MAPK. This activation is an actin-dependent mechanism that requires internalization of live parasites. Once stably infected, macrophages become unresponsive to subsequent parasite infection. Priming of cells with IFNgamma, prior to Leishmania infection, did not prevent the silencing of MAPKs pathways induced by Leishmania parasites. NF-kappaB transcriptional activity in response to Leishmania infection is also impaired in stably infected cells. This impairment was not due to MAPK deactivation as inhibition of ERK1/2 and p38MAPK, actually enhances the transcriptional activity of NF-kappaB in response to initial contact of Leishmania with the murine macrophagic cell line Raw264.7. Moreover, Leishmania parasites could not reverse the hyporesponsive state induced by LPS. These effects do not reflect a general down-regulation of macrophages signaling by parasites, as cells with established Leishmania infection display normal response to PMA. In addition we show that the mechanisms of Leishmania induced hyporesponsive state is not due to the induction of a cellular tyrosine phosphatase activity as previously reported in LPS treated cells. PMID- 18406465 TI - Differential contribution of neutrophilic granulocytes and macrophages to nitrosative stress in a host-parasite animal model. AB - Tyrosine nitration is a hallmark for nitrosative stress caused by the release of reactive oxygen and nitrogen species by activated macrophages and neutrophilic granulocytes at sites of inflammation and infection. In the first part of the study, we used an informative host-parasite animal model to describe the differential contribution of macrophages and neutrophilic granulocytes to in vivo tissue nitration. To this purpose common carp (Cyprinus carpio) were infected with the extracellular blood parasite Trypanoplasma borreli (Kinetoplastida). After infection, serum nitrite levels significantly increased concurrently to the upregulation of inducible nitric oxide synthase (iNOS) gene expression. Tyrosine nitration, as measured by immunohistochemistry using an anti-nitrotyrosine antibody, dramatically increased in tissues from parasite-infected fish, demonstrating that elevated NO production during T. borreli infection coincides with nitrosative stress in immunologically active tissues. The combined use of an anti-nitrotyrosine antibody with a panel of monoclonal antibodies specific for several carp leukocytes, revealed that fish neutrophilic granulocytes strongly contribute to in vivo tissue nitration most likely through both, a peroxynitrite- and an MPO-mediated mechanism. Conversely, fish macrophages, by restricting the presence of radicals and enzymes to their intraphagosomal compartment, contribute to a much lesser extent to in vivo tissue nitration. In the second part of the study, we examined the effects of nitrosative stress on the parasite itself. Peroxynitrite, but not NO donor substances, exerted strong cytotoxicity on the parasite in vitro. In vivo, however, nitration of T. borreli was limited if not absent despite the presence of parasites in highly nitrated tissue areas. Further, we investigated parasite susceptibility to the human anti-trypanosome drug Melarsoprol (Arsobal), which directly interferes with the parasite-specific trypanothione anti-oxidant system. Arsobal treatment strongly decreased T. borreli viability both, in vitro and in vivo. All together, our data suggest an evolutionary conservation in modern bony fish of the function of neutrophilic granulocytes and macrophages in the nitration process and support the common carp as a suitable animal model for investigations on nitrosative stress in host parasite interactions. The potential of T. borreli to serve as an alternative tool for pharmacological studies on human anti-trypanosome drugs is discussed. PMID- 18406466 TI - Executive attention and self-regulation in infancy. AB - This study investigates early executive attention in infancy by studying the relations between infant sequential looking and other behaviors predictive of later self-regulation. One early marker of executive attention development is anticipatory looking, the act of looking to the location of a target prior to its appearance in that location, a process that involves endogenous control of visual orienting. Previous studies have shown that anticipatory looking is positively related to executive attention as assessed by the ability to resolve spatial conflict in 3-4-year-old children. In the current study, anticipatory looking was positively related to cautious behavioral approach in response to non-threatening novel objects in 6- and 7-month-old infants. This finding and previous findings showing the presence of error detection in infancy are consistent with the hypothesis that there is some degree of executive attention in the first year of life. Anticipatory looking was also related to the frequency of distress, to looking away from disturbing stimuli, and to some self-regulatory behaviors. These results may indicate either early attentional regulation of emotion or close relations between early developing fear and later self-regulation. Overall, the results suggest the presence of rudimentary systems of executive attention in infants and support further studies using anticipatory looking as a measure of individual differences in attention in infancy. PMID- 18406467 TI - The ontogeny of drug metabolism enzymes and implications for adverse drug events. AB - Profound changes in drug metabolizing enzyme (DME) expression occurs during development that impacts the risk of adverse drug events in the fetus and child. A review of our current knowledge suggests individual hepatic DME ontogeny can be categorized into one of three groups. Some enzymes, e.g., CYP3A7, are expressed at their highest level during the first trimester and either remain at high concentrations or decrease during gestation, but are silenced or expressed at low levels within one to two years after birth. SULT1A1 is an example of the second group of DME. These enzymes are expressed at relatively constant levels throughout gestation and minimal changes are observed postnatally. ADH1C is typical of the third DME group that are not expressed or are expressed at low levels in the fetus, usually during the second or third trimester. Substantial increases in enzyme levels are observed within the first one to two years after birth. Combined with our knowledge of other physiological factors during early life stages, knowledge regarding DME ontogeny has permitted the development of robust physiological based pharmacokinetic models and an improved capability to predict drug disposition in pediatric patients. This review will provide an overview of DME developmental expression patterns and discuss some implications of the data with regards to drug therapy. Common themes emerging from our current knowledge also will be discussed. Finally, the review will highlight gaps in knowledge that will be important to advance this field. PMID- 18406468 TI - Specific detection of CB1 receptors; cannabinoid CB1 receptor antibodies are not all created equal! AB - The study of endogenous cannabinoid CB1 receptor proteins in neuronal tissues and cells relies on the availability of highly specific antibodies. We have tested the ability of a series of CB1 antibodies to detect endogenous receptors in brain as well as hemagglutinin (HA)-tagged receptors transfected into HEK-293 cells using a combination of immunological methods. An initial comparison of several CB1 antibodies in mouse brain revealed substantial differences in staining pattern to ligand binding by autoradiography. Antibodies were then tested immunocytochemically against HEK cells expressing HA-tagged rat and human CB1 receptors. None of the commercial antibodies tested were able to detect the receptor in this context. All antibodies were then screened by Western blotting using lysates from the HEK cells and rodent brain homogenates. Again, none of the commercially available antibodies detected proteins of the correct molecular weight in transfected cell lines or brain homogenates, although all recognized multiple proteins in brain tissues. We conclude that the commercially available antibodies we tested failed to detect CB1 receptors abundantly expressed in HEK cells or native receptors in brain slices or homogenates. As such, comprehensive validation of the specificity of these CB1 antibodies for a particular application is essential before use. PMID- 18406469 TI - Comparison of clinical characteristics, co-morbidity and pharmacotherapy in adolescent schizophrenia patients with and without obsessive-compulsive disorder. AB - A substantial proportion of adolescent schizophrenia patients exhibit obsessive compulsive symptoms/disorder (OCS/OCD). In the present study we sought to provide a clinical characterization of adolescent schizo-obsessive patients. A consecutive sample of 22 adolescent patients (age 13-18 years) who met DSM-IV criteria for both schizophrenia and OCD was compared with 22 non-OCD schizophrenia patients matched for age, gender and number of hospitalizations. The Structured Clinical Interview for DSM-IV Axis I psychiatric disorders (SCID I), the Scale for the Assessment of Positive (SAPS) and Negative (SANS) Symptoms, the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) and the Clinical Global Impression (CGI) were used. We found that schizo-obsessive patients had earlier age at onset of schizophrenia symptoms, had more OCD spectrum disorders, primarily tic disorders, but did not differ in severity of schizophrenia symptoms from non-OCD schizophrenia patients. In a majority of the schizo-obsessive patients, OCS preceded or co-occurred with the onset of schizophrenia and did not correlate with schizophrenic symptoms. As expected, more schizo-obsessive patients than their non-OCD counterparts were treated with adjunctive anti obsessive agents. These findings indicate that clinical characteristics of adolescent schizo-obsessive patients are generally similar to those previously revealed in their adult counterparts. The neurobiology underlying the co occurrence of the OC and schizophrenia symptoms merits further evaluation. PMID- 18406470 TI - High iNOS expression in macrophages in canine leishmaniasis is associated with low intracellular parasite burden. AB - The expression of iNOS by macrophages in 33 dogs suffering from spontaneous leishmaniasis was analysed by immunohistochemistry in skin, liver and lymph nodes. A correlation study between the number of macrophages expressing iNOS and the number of macrophages containing leishmania amastigotes was carried out. Formalin-fixed paraffin-embedded tissue sections from the skin (28 cases), popliteal lymph nodes (8 cases) and liver (3 cases) of dogs of different age, sex and breed suffering from leishmaniasis were included in the study. Dogs were referred as positive for Leishmania spp by serology and the diagnosis was confirmed by demonstration of leishmania amastigotes within macrophages by histopathology. Tissue samples of skin (3 cases), popliteal lymph nodes (5 cases) and liver (3 cases) from dogs seronegative for leishmaniasis with no histopathological changes were included in the study as controls. The immunohistochemical study revealed that macrophages containing a high number of leishmania did not express iNOS. Correlation between the number of macrophages expressing iNOS and the number of macrophages containing leishmania amastigotes was assessed using the Spearman test. High expression of iNOS in macrophages was related with low number of leishmania amastigotes in macrophages in all cases (r= 0.47, p=0.002). These results suggest that iNOS expression by macrophages plays an important role during the control of Leishmania infection in dogs. PMID- 18406471 TI - Assessment of in vitro interferon-gamma responses from peripheral blood mononuclear cells of cattle infected with Mycoplasma mycoides ssp. mycoides small colony type. AB - Contagious bovine pleuropneumonia (CBPP) is a lung disease caused by the bacterial pathogen Mycoplasma mycoides ssp. mycoides small colony type (MmmSC). It has been spreading due to a number of factors including poor vaccine efficacy and poor sensitivity of current diagnostic tests. The purpose of this study was to assess interferon gamma (IFN-gamma) release after stimulation of peripheral blood mononuclear cells (PBMC) from experimentally infected cattle. PBMC collected from 15 artificially infected animals were incubated with different concentrations of total MmmSC antigen. After 72h of incubation the IFN-gamma release was measured and found to be elevated in 11 animals. We did not observe a correlation between IFN-gamma release of animals with and without pathomorphological gross lesions. Therefore, our data do not confirm a role for CD4 T-lymphocytes in protection, since there is no correlation between IFN-g secretion (supposed to be mainly derived from CD4 T-cells) and disease severity. Additionally, we applied immunocytochemistry on affected lung tissue and detected no build up of T-lymphocytes (CD4 T-cells, CD8 T-cells) but a high presence of myeloid cells. PMID- 18406472 TI - Microglial Fc receptors mediate physiological changes resulting from antibody cross-linking of myelin oligodendrocyte glycoprotein. AB - Antibodies to myelin oligodendrocyte glycoprotein (MOG) have been implicated in Multiple Sclerosis demyelination through activation of complement and/or macrophage-effector processes. We presented a novel mechanism, whereby MOG on oligodendrocytes, when cross-linked with anti-MOG and secondary antibody resulted in its repartitioning into lipid rafts, and changes in protein phosphorylation and morphology. Here, we show that similar events occur when anti-MOG is cross linked with Fc receptors (FcRs) present on microglia but not with complement. These results indicate that FcRs are endogenous antigen/antibody cross-linkers in vitro, suggesting that FcRs could be physiologically relevant in vivo and possible targets for therapy in Multiple Sclerosis. PMID- 18406473 TI - Early blood-brain barrier permeability in cerebella of PLSJL mice immunized with myelin basic protein. AB - The blood-brain barrier (BBB) is dramatically but transiently compromised in the cerebella of myelin basic protein immunized mice at least 1 week prior to the development of the paralytic phase of experimental allergic encephalomyelitis (EAE). Treatment of mice with the peroxynitrite-dependent radical scavenger uric acid (UA) during the first week after immunization blocks the early increase in cerebellar BBB permeability and the subsequent development of clinical signs of EAE. These results indicate that the early loss of BBB integrity in the cerebellum is likely to be a necessary step in the development of paralytic EAE. PMID- 18406474 TI - Bickerstaff's brainstem encephalitis after an outbreak of Campylobacter jejuni enteritis. AB - Twenty-eight patients suffered Campylobacter jejuni enteritis after eating raw chicken. Among them, only one patient developed Bickerstaff's brainstem encephalitis, who carried anti-GQ1b IgG antibodies. In contrast, none of the others did the autoantibodies. C. jejuni was cultured from all stool samples from five patients with enteritis alone. All the isolates had the same genotype, cst II (Asn51), which are characteristic of strains isolated from Bickerstaff's brainstem encephalitis. These findings suggest that host susceptibility may play a role in inducing the production of anti-ganglioside antibodies and the development of Bickerstaff's brainstem encephalitis. PMID- 18406475 TI - Expression of the capsid protein of porcine circovirus type 2 in Lactococcus lactis for oral vaccination. AB - Diseases associated with porcine circovirus type 2 (PCV2) infections are becoming a major problem for the swine industry worldwide. The capsid protein (Cap) of PCV2 is an antigen important for both early diagnosis and development of vaccines. In this study, Lactococcus lactis was used as vehicle to deliver the PCV2 antigen in an attempt to develop oral vaccine. A cap gene with a deleted nuclear localization signal sequence (dcap) was cloned into an Escherichia coli/L. lactis shuttle vector pSEC: LEISS under the control of a nisin promoter. Intracellular and extracellular expression of the dCap was confirmed by Western blot analysis. Significantly higher levels of PCV2-specific IgG in the sera of mice were observed upon oral administration of strain cultures expressing the PCV2 antigen. These results suggest that it is feasible to use L. lactis as an antigen delivery vehicle for developing oral vaccines against PCV2 infection. PMID- 18406476 TI - Mentalizing in economic decision-making. AB - In the Ultimatum Game, participants typically reject monetary offers they consider unfair even if the alternative is to gain no money at all. In the present study, ERPs were recorded while subjects processed different offers of a proposer. In addition to clearly fair and unfair offers, mid-value offers which cannot be easily classified as fair or unfair and therefore involve more elaborate decision making were analyzed. A fast initial distinction between fair and other kinds of offers was reflected by amplitude of the feedback related negativity (FRN). Mid-value offers were associated with longer RTs, and a larger N350 amplitude. In addition, source analyses revealed a specific involvement of the superior temporal gyrus and the inferior parietal lobule during processing of mid-value offers compared to offers categorized clearly as fair or unfair, suggesting a contribution of mentalizing about the intention of the proposer to the decision making process. Taken together, the present findings support the idea that economic decisions are significantly affected by non-rational factors, trying to narrow the gap between formal theory and the real decisional behaviour. PMID- 18406477 TI - The effects of maternal Cd on the metallothionein expression in tilapia (Oreochromis mossambicus) embryos and larvae. AB - The purpose of this study was to identify the factor(s) which would enhance the Cd resistance as assessed by the metallothionein (MT) expression in tilapia larvae. Larvae were collected from parents that were pretreated respectively with Cd or saline. At the end of the 12-week experiment, the hepatic MT and Cd contents in the breeding female fish were recorded. Our results indicated that a significant relationship between Cd and MT contents can be found in the offspring from the parent fish treated with Cd. However, the higher Cd resistance, Cd contents, and MT expression were limited to those larvae from parent fish bred within 4 weeks of the injection. By week 12, the Cd-treated fish still contained high levels of MT in their hepatic tissues. However, the MT and Cd contents in the larvae from these adult fish were not significantly different from those from the controls. In summary, we suggest that the higher Cd resistance of larvae from the egg stage was a result of the Cd contamination of the parent female, as evidenced by an increase in MT expression induced in tilapia embryos and larvae. PMID- 18406478 TI - Programmed Cell Death 5 from Toxoplasma gondii: a secreted molecule that exerts a pro-apoptotic effect on host cells. AB - Although parasite-infected host cells become resistant to apoptosis, uninfected bystander cells undergo apoptosis during Toxoplasma gondii infection. The Programmed Cell Death 5 (TgPDCD5) gene, a homologue of the human apoptosis related molecule, was cloned from a T. gondii full-length cDNA database and subsequently characterized. The native TgPDCD5 was located in the cytosol and also detected in the secreted fraction. Immuno-electron microscopic analysis showed TgPDCD5 was primarily located close to the rhoptries or vesicle-like structures near the surface membrane of the parasite. Studies using recombinant TgPDCD5 (rTgPDCD5) demonstrated that host cells internalize the molecule in a heparan sulfate proteoglycan-binding motif-dependent manner. Furthermore, the addition of rTgPDCD5 to culture medium resulted in the enhancement of host-cell apoptosis triggered by etoposide in macrophage cell line J774A.1 and leukemic cell line HL-60 cells. Additionally, rTgPDCD5 induced apoptosis in J774A.1 cells in the presence of IFN-gamma. This report is the first to identify a parasitic molecule of T. gondii that has a pro-apoptotic effect on host cells. PMID- 18406479 TI - Relationship between protein and mitochondrial DNA oxidative injury and telomere length and muscle loss in healthy elderly subjects. AB - A blood sample and muscle biopsies were obtained from 54 elderly subjects. Twenty seven subjects aged 77+/-3 years, had experienced a change in fat free mass (FFM) of +194+/-282g/year (lean body mass maintainers) and 27 subjects aged 78+/-3 years, had a change in FFM of -487+/-209g/year (lean body mass losers). Muscle biopsies were also obtained from 10 healthy subjects aged 34+/-4 years. In muscle, the ratio of mitochondrial DNA (mtDNA) to nuclear DNA (nDNA) and telomere length were assessed and deposition of 4-hydroxy-2-nonenal adducts (4HNE) was visualized by electron microscopy. In FFM maintainers, losers and young controls, the ratio of mtDNA to nDNA was 2.1 (95% confidence intervals (CI), 0.1-31.7), 1.5 (95% CI, 0.2-15.7) and 18.6 (95% CI, 2.8-46.2), respectively. 4HNE deposition was 5.9 (95% CI, 1.5-28), 4.9 (95% CI, 0.9-13) and 3.4 (95% CI, 1.1-4.6) gold particles/microm(2), respectively. Telomere length, expressed as T/S ratio, was 0.06 (95% CI, 0.01-0.16), 0.06 (95% CI, 0.03-0.27) and 0.34 (95% CI, 0.1-1.34), respectively (p<0.02 or less for all comparisons between elderly and young subjects). PMID- 18406480 TI - Acute stress-induced cardiomyopathy: a brief observation. AB - Acute stress-induced cardiomyopathy is a syndrome originally described in Japan but increasingly recognized all over the world. Patients typically present with chest pain triggered by an intense emotional or physiological stressor, electrocardiographic changes, and mild cardiac enzyme elevation. Patients typically return to normal cardiac function within 1 month. A retrospective review was conducted of all acute stress-induced cardiomyopathy cases at the University of Maryland Medical Center between May 2006 and May 2007. Six patients fulfilled the criteria and were included in the study. The main clinical features of the syndrome were statistically analyzed. Most patients were post-menopausal women presenting with chest pain. An emotional or physiological stressor was identified in 3 of the 6 patients. Electrocardiographic changes (ST segment elevation or T wave inversion) were present in 3 patients. All patients had left ventricular apical hypokinesis with basal hyperkinesis and elevated cardiac enzymes. The 3 patients receiving a follow up echocardiogram had improved left ventricular contractility and reduced mitral regurgitation within 1 month. Two of the patients had complete resolution in 3-4 days. Stress-induced cardiomyopathy is an important syndrome which mimics an acute myocardial infarction. It is typically self-limiting but can result in ventricular arrhythmias and cardiogenic shock. Our findings should caution clinicians that an identifiable stressor and electrocardiographic changes are not always present. In addition, our observation suggests that patients may regain normal cardiac function in less than 4 days. PMID- 18406481 TI - Recurrent left ventricular apical ballooning induced by recurrent stress. AB - This paper reports on a 67-year-old woman with apical ballooning and recurrent dyskinesia of left ventricular wall. During MRI-examination the patient developed severe apical dyskinesia of the left ventricle induced by agoraphobia in the magnet. PMID- 18406482 TI - Factors associated with endemic raccoon (Procyon lotor) rabies in terrestrial mammals in New York State, USA. AB - This study evaluated characteristics associated with raccoon (Procyon lotor) rabies in New York State (NYS), USA, where this disease has been endemic for the last 15 years. The study included 4448 cases of raccoon rabies in terrestrial mammals reported across 1639 census tracts of NYS during 1997-2003. A Poisson regression model with census tract-year as the unit of analysis revealed a higher number of raccoon-variant rabies cases per square kilometer in census tracts with each percent increase in the proportion of low-intensity residential areas (those with a lower concentration of housing units) (RR=7.68) and a lack of rivers/lakes (RR=1.20) and major roads (RR=1.10), while the number of cases decreased with each 1-m increase in land elevation (RR=0.998), and each percent increase in the proportion of wetlands (RR=0.01). The model was adjusted for county, ecoregion, and latitude to help control for unknown spatially dependent covariates. The model may be used in prioritizing areas for rabies control based on differential risk, including use of costly intervention methods such as oral rabies vaccine. PMID- 18406483 TI - Comparison of three methods of surveillance with application to the detection of Johne's disease seropositivity in Texas cattle. AB - Surveillance and monitoring are important for measuring the level of disease in a population, documenting changes in prevalence over time, determining high-risk areas for implementation of control measures, eradicating disease, and documenting freedom from disease. The documentation of freedom from disease has importance for international trade and the resumption of production after an outbreak. Johne's disease (JD) is an example of an endemic disease of cattle that has variable prevalence related to environmental and animal-level factors. Three methods of sample collection were used to describe the prevalence and distribution of JD seropositivity in Texas. Sampled cattle were: (1) extensively managed herds, (2) market cattle, and (3) clinically ill cattle examined by practicing veterinarians throughout Texas. Samples were evaluated for JD using a commercially available serum ELISA. Proportion of seropositive samples was compared and spatial distributions were evaluated for clustering. Difference of JD seropositivity was observed among the three sample populations suggesting that estimation of disease prevalence is dependant upon the source of samples. PMID- 18406484 TI - Moral hazard and the demand for health services: a matching estimator approach. AB - We estimate the impact of extra health insurance coverage beyond a National Health System on the demand for several health services. Traditionally, the literature has tried to deal with the endogeneity of the private (extra) insurance decision by finding instrumental variables. Since a priori instrumental variables are hard to find we take a different approach. We focus on the most common health insurance plan in Portugal, ADSE, which is given to all civil servants and their dependents. We argue that this insurance is exogenous, i.e., not correlated with the beneficiaries' health status. This identifying assumption allows us to estimate the impact of having ADSE coverage on the demand for three different health services using a matching estimator technique. The health services used are number of visits, number of blood and urine tests, and the probability of visiting a dentist. Results show large positive effects of ADSE coverage for number of visits and tests among the young (18-30 years old) but only the latter is statistically significantly different from zero. The effects represent 21.8% and 30% of the average number of visits and tests for the young. On the contrary, we find no evidence of moral hazard on the probability of visiting a dentist. PMID- 18406485 TI - The effect of startle reflex habituation on cardiac defense: interference between two protective reflexes. AB - The present study investigated the relationship between blink startle and cardiac defense, two protective reflexes that are said to be elicited by the transient and the sustained components, respectively, of high intensity stimuli. Three groups of participants were presented with three intense long lasting noise stimuli (500ms) after habituation training with 12 brief (50ms) high intensity noise bursts (High group), low intensity noise bursts (Low group) or high intensity visual stimuli (Light group). The transition from habituation to defense stimuli resulted in increased blink startles in groups Low and Light, but not in group High. A cardiac defense reflex, characterised by a short and long delayed increase in heart rate, was observed in group Light, but not in groups Low and High. This pattern of results indicates that habituation to startle eliciting stimuli will impair defense reflexes elicited on subsequent test trials and suggests some interrelation between the two reflex systems. PMID- 18406486 TI - Constrained action selection in children with developmental coordination disorder. AB - The effect of advance ('precue') information on short aiming movements was explored in adults, high school children, and primary school children with and without developmental coordination disorder (n=10, 14, 16, 10, respectively). Reaction times in the DCD group were longer than in the other groups and were more influenced by the extent to which the precue constrained the possible action space. In contrast, reaction time did not alter as a function of precue condition in adults. Children with DCD showed greater inaccuracy of response (despite the increased RT). We suggest that the different precue effects reflect differences in the relative benefits of priming an action prior to definitive information about the movement goal. The benefits are an interacting function of the task and the skill level of the individual. Our experiment shows that children with DCD gain a benefit from advance preparation in simple aiming movements, highlighting their low skill levels. This result suggests that goal-directed RTs may have diagnostic potential within the clinic. PMID- 18406487 TI - Lactic acid increases aquaporin 4 expression on the cell membrane of cultured rat astrocytes. AB - The water channel protein aquaporin (AQP) may play roles in the homeostasis of water content in the brain and brain edema. One possible mechanism of brain edema is glial swelling due to lactic acidosis associated with ischemia. Here, we investigated the effect of lactic acid on the expression and cellular distribution of AQP 4 in cultured rat astrocytes. After 24h of incubation, the AQP4 expression level increased maximally with 35mM lactic acid. The AQP4 expression levels also increased with hydrochloric acid or acetic acid. In contrast, with sodium lactate, the AQP4 levels did not increase. The increase in AQP4 expression level occurred without a significant increase in AQP4 mRNA expression level by lactic acid. Under the conditions of de novo protein synthesis inhibition with cycloheximide, lactic acid increased the AQP4 expression level. Furthermore, lactic acid increased the AQP4 expression level on the cell surface of the astrocytes, as determined by a cell surface biotinylation assay and immunocytochemical examination. The increase in AQP4 expression level on the cell membrane of astrocytes induced by lactic acid may be a new regulation mechanism of AQP4 in the brain. PMID- 18406488 TI - Complete genome sequence analysis of dengue virus type 2 isolated in Brunei. AB - In a previous study, we have reported the detection and isolation of dengue virus in Brunei (Osman, O., Fong, M.Y., Devi, S., 2007. A preliminary study of dengue infection in Brunei. JJID 60 (4), 205-208). DEN-2 was the predominant serotype followed by DEN-1. The full genomic sequences of 3 DEN-2 viruses isolated during the 2005-2006 dengue incident in Brunei were determined. Twenty-five primer sets were designed to amplify contiguous overlapping fragments of approximately 500 600 base pairs spanning the entire sequence of the viral genome. The amplified PCR products were sent for sequencing and their nucleotides and the deduced amino acids were determined. All three DEN-2 virus isolated were clustered in the Cosmopolitan genotype of the DEN-2 classification by Twiddy et al. This work constitutes the first complete genetic characterization of three Brunei DEN-2 virus strains. PMID- 18406489 TI - Molecular interactions, internal structure and drug release kinetics of rationally developed polymer-lipid hybrid nanoparticles. AB - This paper presents the first study of molecular interactions of ingredients and internal nanostructure in relation to drug loading and release mechanisms/kinetics of rationally designed solid polymer-lipid hybrid nanoparticles (PLN). The PLN were prepared by using a rationally selected composition that was found in our previous work to provide optimized interactions of verapamil hydrochloride (VRP) with dextran sulfate sodium (DS) and then the VRP-DS complex with dodecanoic acid (DA). The solid-state properties of the components, their molecular interactions and the morphology, particle size and internal structure of PLN were determined by use of differential scanning calorimetry, powder X-ray diffraction, (13)C nuclear magnetic resonance, Fourier transform infrared spectroscopy, transmission electron microscopy (TEM) and dynamic light scattering. The distribution of VRP in PLN was examined by TEM imaging using a cationic gold tracer. Drug release studies were conducted in various media. Drug loading as high as 36% and loading efficiencies up to 99% were achieved in the rationally formulated PLN. Hydrogen bonding between drug, polymer and lipid and a uniform distribution of amorphous VRP within the solid lipid matrix were evident. Sustained drug release from the PLN was mainly controlled by ion exchange and diffusion processes. The results demonstrated that strong molecular interactions among the drug, the polymer and the lipid in the optimized formulation were responsible for the improved drug loading and release performance of the PLN. PMID- 18406490 TI - Epileptic EEG detection using neural networks and post-classification. AB - Electroencephalogram (EEG) has established itself as an important means of identifying and analyzing epileptic seizure activity in humans. In most cases, identification of the epileptic EEG signal is done manually by skilled professionals, who are small in number. In this paper, we try to automate the detection process. We use wavelet transform for feature extraction and obtain statistical parameters from the decomposed wavelet coefficients. A feed-forward backpropagating artificial neural network (ANN) is used for the classification. We use genetic algorithm for choosing the training set and also implement a post classification stage using harmonic weights to increase the accuracy. Average specificity of 99.19%, sensitivity of 91.29% and selectivity of 91.14% are obtained. PMID- 18406491 TI - Efficient construction of therapeutics, bioconjugates, biomaterials and bioactive surfaces using azide-alkyne "click" chemistry. AB - The concept of "click" chemistry, introduced by Sharpless and coworkers a couple of years ago, promotes the use of efficient, selective and versatile chemical reactions in synthetic chemistry. For instance, the copper(I)-catalyzed azide alkyne cycloaddition (CuAAC) is regarded as a prime example of "click" chemistry. This reaction is regioselective, chemoselective and moreover can be performed in aqueous medium at room or physiological temperature. Thus, CuAAC became lately a very popular ligation tool in biological and medical sciences. Several hundred of articles exploring the synthetic possibilities of CuAAC in biosciences have been published within the last four years. The aim of the present review is to give an overall--non exhaustive--picture of this emerging field of research. The advantages and versatility of CuAAC in scientific disciplines as diverse as drug discovery, biochemistry, bioconjugates synthesis, drug-delivery, gene therapy, bioseparation or diagnostics are presented and discussed in detail. PMID- 18406493 TI - Nickel release from earrings purchased in the United States: the San Francisco earring study. AB - BACKGROUND: Nickel sensitization is frequent among US patients with dermatitis and in the general population. In Europe, decreasing prevalences of nickel sensitization are observed as a result of the European Union Nickel Directive. However, no directive exists in the United States. OBJECTIVES: We sought to examine nickel release from inexpensive earrings and discuss possible methods of reducing nickel exposure among consumers. METHODS: A total of 34 different stores and artists were visited. A total of 277 earrings were purchased and examined with the dimethylglyoxime (DMG) test. RESULTS: DMG-positive earrings were identified from local artists (69%) and tourist stores (42.9%) but also chain stores targeting young (24.1%) and mature (1.7%) women. No correlation between price and outcome of DMG testing was identified. LIMITATIONS: Our study does not reflect buying trends in the United States. Pretreatment of earrings with artificial sweat was not performed and the DMG may lead to both false-positive and false-negative results. CONCLUSIONS: Nickel exposure from inexpensive earrings is frequent in the United States. A regulatory governmental intervention may be warranted. PMID- 18406492 TI - Components of a successful intervention for monthly skin self-examination for early detection of melanoma: the "Check It Out" trial. AB - BACKGROUND: Multicomponent Check-It-Out project interventions were found to be effective at increasing thorough skin self-examinations (TSSE). Application of this research will benefit from determining the most important interventions for promoting TSSE. OBJECTIVE: Identification of the most important Check-It-Out intervention components for promoting TSSE. METHODS: This study analyzed the responses of the 567 participants who were randomized into the skin examination intervention group and who did not report performing TSSE at their baseline interview. RESULTS: Watching the video, using the hand mirror, shower card, American Cancer Society brochure, sample photographs, and finding the health educator helpful were associated with performing TSSE at 2 months, 12 months, or both. LIMITATIONS: Use of the materials within the skin group was not randomly assigned. All data were based on participant self-report. CONCLUSIONS: As future interventions to increase TSSE are developed, the identified useful components will be important to include. PMID- 18406494 TI - Overweight and obesity: the role of education, employment and income in Spanish adults. AB - OBJECTIVE: To study the relationship between obesity prevalence and education, employment and socioeconomic levels to improve the designing of intervention programs. METHODS: Cross-sectional population-based study on 2640 subjects over 15 years of age in Spain. Through interviews conducted in subjects' homes we have obtained socio-demographic, anthropometric data, and information on physical activity, life styles and eating habits. Statistically, we have used prevalence ratios with confidence intervals and logistic regressions. RESULTS: The prevalence of overweight and obesity is 36.4% (IC95% 34.5-38.2) and 17% (IC95% 15.7-18.5), respectively, higher in men, and increasing with age. Obesity prevalence is independent of education level. Higher prevalence of overweight and obesity are found in the lowest occupational categories, and there is an inverse relationship between BMI and employment situations. Obesity is more prevalent in retired people and people that work from home, compared with professions that require activity at work. The association found between obesity and socioeconomic characteristics of the population is different in men and women: in women only, this risk was found to be associated with low or primary education levels (OR 2.4, 1.5-4.0), being unemployed or working at home (OR 1.6, 1.08-2.4) and having a medium income (OR 1.87, 1.03-3.33). CONCLUSIONS: We have confirmed a high prevalence of overweight and obesity in our area. Socioeconomic variables have a more predictive value in women than in men. PMID- 18406495 TI - Structure-activity relations of parasin I, a histone H2A-derived antimicrobial peptide. AB - The structure-activity relations and mechanism of action of parasin I, a 19-amino acid histone H2A-derived antimicrobial peptide, were investigated. Parasin I formed an amphipathic alpha-helical structure (residues 9-17) flanked by two random coil regions (residues 1-8 and 18-19) in helix-promoting environments. Deletion of the lysine residue at the N-terminal [Pa(2-19)] resulted in loss of antimicrobial activity, but did not affect the alpha-helical content of the peptide. The antimicrobial activity was recovered when the lysine residue was substituted with another basic residue, arginine ([R(1)]Pa), but not with polar, neutral, or acidic residues. Progressive deletions from the C-terminal [Pa(1-17), Pa(1-15)] slightly increased the antimicrobial activity (1-4 microg/ml) without affecting the alpha-helical content of the peptide. However, further deletion [Pa(1-14)] resulted in nearly complete loss of antimicrobial activity and alpha helical structure. Confocal microscopic analysis and membrane permeabilization assays showed that parasin I and its analogs with comparable antimicrobial activities localized to the cell membrane and subsequently permeabilized the outer and cytoplasmic membranes. Pa(1-14) also localized to the cell membrane, but lost membrane-permeabilizing activity, whereas Pa(2-19) showed poor membrane binding and -permeabilizing activities. The results indicate that the basic residue at the N-terminal is essential for the membrane-binding activity of parasin I, and among the membrane-binding parasin I analogs, the alpha-helical structure is necessary for the membrane-permeabilizing activity. PMID- 18406496 TI - Developmental changes in the response of murine cerebellar granule cells to nitric oxide. AB - Nitric oxide is a diffusible messenger that plays a multitude of roles within the nervous system including modulation of cell viability. However, its role in regulating neuronal survival during a defined period of neurodevelopment has never been investigated. We discovered that expression of the messenger RNA for both neuronal and endothelial nitric oxide synthase increased in the early postnatal period in the cerebellum in vivo, whilst the expression of inducible nitric oxide synthase remained constant throughout this time in development. Whilst scavenging of nitric oxide was deleterious to the survival of early postnatal cerebellar granule neurons in vitro, this effect was lost in cultures derived at increasing postnatal ages. Conversely, sensitivity to exogenous nitric oxide increased with advancing postnatal age. Thus, we have shown that as postnatal development proceeds, cerebellar granule cells alter their in vitro survival responses to both nitric oxide inhibition and donation, revealing that the nitric oxide's effects on developing neurons vary with the stage of development studied. These findings have important consequences for our understanding of the role of nitric oxide during neuronal development. PMID- 18406497 TI - Indanesulfonamides as carbonic anhydrase inhibitors and anticonvulsant agents: structure-activity relationship and pharmacological evaluation. AB - A small library of indanesulfonamides was screened for the inhibition of the human carbonic anhydrase (CA, EC 4.2.1.1) isoforms involved in neuronal excitation, that is, isoforms VII, XII and XIV. These CA isoforms are becoming interesting target for the design of agents useful for the treatment of epilepsy. The inhibition pattern of these indanesulfonamide compounds towards these three isoforms was excellent, with many nanomolar inhibitors detected (K(I)s in the range of 0.78-10 nM against hCA VII; 0.32-56 nM against hCA XII, and 0.47-1030 nM against hCA XIV, respectively). The maximal electroshock seizure (MES) test performed on mice showed a good anticonvulsant activity for some compounds which protected the mice against convulsions in the 50-62.5% range at a dose of 50 mg/kg. In parallel, the blood-brain barrier passive permeation of these sulfonamides was also estimated by using a computational approach. PMID- 18406498 TI - [Vitamin D2 or vitamin D3?]. AB - PURPOSE: Nearly one billion people around the world are deficient in vitamin D and need to be supplemented. Vitamin D is available in medicines and fortified foods. It is available in two forms: vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol). KEY POINTS: The pharmacopeiae consider these steroid hormones as equivalent and interchangeable. However, several studies have showed that serum level of 25(OH)D is increased more effectively with vitamin D3 than vitamin D2. Vitamin D2 has shorter plasma half-life and a lower affinity for the vitamin D binding protein, the hepatic vitamin D hydroxylase and the vitamin D receptor. CONCLUSION: Vitamin D2 should not be regarded anymore as suitable for supplementation or fortification. Currently though, it is still the most used in some countries such as Portugal and Australia. PMID- 18406499 TI - Elective termination of pregnancy after vaccination reported to the Vaccine Adverse Event Reporting System (VAERS): 1990-2006. AB - Generally, live-virus vaccines are contraindicated for pregnant women because of the theoretical risk of transmission of the vaccine virus to the fetus. Advisory groups recommend avoiding pregnancy in the immediate period after administration of such contraindicated vaccines (CVs) and stress benefit-to-risk evaluation for live or inactivated vaccines regarding pregnancy. Given the limited available data and theoretical risks associated particularly with live-virus vaccines, inadvertent immunization with CVs may lead to elective termination of pregnancy (ETP), despite advisory group statements that "vaccination is not ordinarily an indication to terminate the pregnancy." The Vaccine Adverse Event Reporting System (VAERS), a national passive surveillance system managed by the Food and Drug Administration (FDA) and Centers for Disease Control and Prevention (CDC), accepts reports of adverse events after vaccination. The objectives of this review were to describe reports of ETP in VAERS and characterize the circumstances of inadvertent administration of vaccines to pregnant women among ETP reports. We reviewed VAERS reports of ETP submitted from 1990 to 2006. Reports of ETP for reasons other than vaccination during or shortly before pregnancy, such as fetal abnormalities or deaths, were excluded. Of 80 ETP reports, 62 (78%) originated from the US; 79 (99%) were reported by manufacturers. Median age of vaccinees was 26 years (range: 13-43 years; 67 reports). Seventy-three vaccinees (91%) received a single vaccine; 65 (81%) received at least one live-virus vaccine. In 48 (60%) ETP reports, vaccinees were unaware of pregnancy at time of immunization. In 15 (19%) reports, vaccinees became pregnant within 3 months of vaccination; in 13 (16%) reports, vaccinees might have been pregnant before vaccination; in 4 (5%) reports, information was missing. All 80 reports of ETP involved vaccines for which possible effects on fetal development are unknown. However, no cases of vaccine-associated congenital rubella or varicella syndromes have been reported in the medical literature. Also, these syndromes have not been reported to varicella or rubella vaccine pregnancy registries. VAERS has the limitations of passive surveillance systems. Under-reporting of ETP in VAERS could be substantial. More attention may be needed to assess the likelihood of pregnancy when administering vaccines to women with child-bearing potential, and to inform women who learn they are pregnant shortly after being immunized of current information on risks. Quantifying the frequency of ETP related to CVs and the risk (if any) to the fetus of such vaccines can help to inform policy, practice, and individual decision making. Good quality information may be obtained from controlled observational studies. PMID- 18406500 TI - The effect of the new "24 hour alcohol licensing law" on the incidence of facial trauma in London. AB - On 24 November 2005 the new 2003 Licensing Act was implemented. It permits licensed premises to close at different times under English and Welsh law, rather than at 2300h as under the previous law. The aim of this study was to assess whether head and neck trauma secondary to alcohol-associated assaults had increased, decreased, or stayed the same since the introduction of the act. Data were collected from the Accident and Emergency Department, University College Hospital, attendance databases for two six-month periods: 24 November 2004 to 30 April 2005, and 24 November 2005 to 30 April 2006. There were 1102 attendances for head and neck trauma secondary to alcohol-associated assaults during the six months before the introduction of the 2003 Licensing Act and 730 such attendances during the similar period after the introduction of the law, with fewer cases in each corresponding month during the later period. There were more cases at weekends than on weekdays during both periods. There were fewer cases but more at weekends in 2005-6 than in 2004-5 (423, 58% compared with 584, 53%, respectively). Neither rainfall nor temperature had any influence on the results. The 2003 licensing Act seems to have reduced the number of attendances at the A&E department for head and neck trauma secondary to alcohol associated assaults. PMID- 18406501 TI - Effect of using RNA interference to alter iNOS gene expression on the proliferation of tongue squamous cell carcinoma cell line Tca8113. AB - This study used RNA interference (RNAi) to explore the effect of NO and inducible nitric oxide synthase (iNOS) on apoptosis and proliferation in the tongue squamous carcinoma cell line Tca8113. Tca8113 cells were transfected with the plasmid pGenesil-1, which expresses iNOS short hairpin RNA (shRNA), or the negative control plasmid pSilencer-HK, and the transfected cells were compared with untransfected cells. The expression of iNOS was detected by histochemistry, and apoptosis was detected by flow cytometry. The expression of iNOS was significantly lower in the pSilencer-iNOS group than in the pSilencer-HK and empty control groups. The apoptosis rate was significantly higher in the pSilencer-iNOS group than in the pSilencer-HK and empty control groups. Growth monitoring showed that proliferation was also inhibited in cells transfected with pSilencer-iNOS. RNAi gene silencing decreased iNOS gene expression, induced apoptosis, and suppressed proliferation in Tca8113 cells. PMID- 18406502 TI - The Dermal Sensitisation Threshold- a TTC approach for allergic contact dermatitis. AB - The Threshold of Toxicological Concern (TTC) is a useful concept that is becoming of increasing interest as an addition to the arsenal of tools used for characterising the toxicological risk of human exposure to chemicals. Traditionally used for low level indirect additives, flavours and contaminants in foods, the TTC obviates the need for toxicological testing of chemicals where human exposure is low. Proposals have recently been made for the use of the TTC for low level ingredients in cosmetic and personal care products. However, use of the TTC is only protective for systemic toxicity endpoints, and cannot be used for local endpoints such as contact sensitisation. In this paper a probabilistic analysis of available sensitisation data, similar to that used in the development of the TTC, is presented. The incidence of sensitisers in the world of chemicals was estimated using the ELINCS (European List of Notified Chemical Substances) data set, and a distribution for sensitisation potency was established using a recently published compilation of Local Lymph Node Assay data. From the analysis of these data sets it is concluded that a Dermal Sensitisation Threshold (DST) can be established below which there is no appreciable risk of sensitisation, even for an untested ingredient. Use of a DST would preclude the need for sensitisation testing of ingredients where dermal exposure is sufficiently low. PMID- 18406503 TI - Does neighborhood violence lead to depression among caregivers of children with asthma? AB - Prior studies have related community violence to depression among children, but few studies have examined this relationship among adults. We hypothesized that victimization, awareness, and fear of neighborhood violence would increase the odds of depression among adult caregivers of children with asthma. We surveyed caregivers in the Baltimore Indoor Environment Study of Asthma in Kids (BIESAK), USA. The primary outcome was screening positive for depression on the Center for Epidemiological Studies Depression index. We assessed victimization, awareness, and fear of neighborhood violence, and conducted spatial analysis identifying subject homes within 500 ft of a homicide to validate survey measures of neighborhood violence. A multilevel logistic model with clustering by neighborhood estimated odds ratios and 95% confidence intervals. Survey responses about fear of neighborhood violence were strongly predicted by having a home within 500 ft of a homicide. Of 150 caregivers of children with asthma, 49% were aware of a neighborhood violent event, 36% were fearful of neighborhood violence, 22% reported victimization, and 27% had a homicide within 500 ft of the home. In our multilevel model, fear of violence increased the odds of depression by 6.7. Victimization was associated with a possible trend towards depression, and awareness of neighborhood violence did not increase the odds of depression. Based on our findings, personal experience with neighborhood violence may be more important than simple awareness. Health care workers should consider screening for depression among patients exposed to community violence. PMID- 18406504 TI - Gender differences in unilateral spatial neglect within 24 hours of ischemic stroke. AB - Hemispatial neglect is a common and disabling consequence of stroke. Previous reports examining the relationship between gender and the incidence of unilateral spatial neglect (USN) have included either a large numbers of patients with few neglect tests or small numbers of patients with multiple tests. To determine if USN was more common and/or severe in men or women, we examined a large group of patients (312 right-handed) within 24 hours of acute right hemisphere ischemic stroke. Multiple spatial neglect tasks were used to increase the sensitivity of neglect detection. No differences based upon gender were observed for the prevalence, severity, or a combined task measure of USN. PMID- 18406505 TI - Amphetamine effects in appetitive acquisition depend on the modality of the stimulus rather than its relative validity. AB - Amphetamine has been shown previously to increase the apportioning of associative strength to weak predictors in appetitive Pavlovian conditioning procedures such as latent inhibition and overshadowing. Manipulating the likelihood with which different conditioned stimuli (CSs) predict subsequent delivery of an unconditioned stimulus (UCS) is an alternative method by which the associability of CSs can be influenced. The present experiment tested effects of D-amphetamine (0.5 mg/kg or 1.5 mg/kg administered 15 min prior to conditioning) in appetitive acquisition under partial versus continuous reinforcement of alternative CSs with sucrose pellet UCS delivery. Specifically, male Wistar rats were conditioned to light and tone CSs that were followed by the UCS on 100% or 50% of trials in a cross-over design. It was predicted that amphetamine would disrupt rats' ability to select appropriately the most valid CSs for learning which would be expressed as increased conditioning to weaker, 50% valid CSs. Contrary to prediction, differential responding based on relative validity was preserved under amphetamine, for both light and tone stimuli. Instead, the results showed that responding to light CSs was generally reduced under amphetamine. Conditioning to tone CSs was higher and unaffected by amphetamine. Thus, results demonstrate that amphetamine effects are determined by the properties of the CS used for learning. PMID- 18406506 TI - Pro-inflammatory potential of wood smoke and traffic-derived particles in a monocytic cell line. AB - Lung inflammation is an important process in host defence to inhaled particulate matter. To what extent physicochemical properties of particles from different sources influence their inflammatory potential has not been fully clarified. The aim of this study was to investigate the potential of particles from wood smoke and traffic to induce a release of pro-inflammatory cytokines in the monocytic cell line THP-1. The influence of endotoxin on cytokine release was investigated using the inhibitor polymyxin B sulphate, whereas the responses to native particles, washed particles and their organic extracts were compared to determine the role of the organic fraction. Particles from the two sources showed a similar inflammatory potential, but the response was mediated by different particle characteristics. The organic fraction of wood smoke accounted for the majority of the cytokine release, whereas the response to the traffic-derived particles was in addition influenced by endotoxin and the particle core. The sum of the cytokine release induced by the organic extract and washed particles was lower than that induced by native particles, suggesting that the organic fraction must be adsorbed to the particles to exert biological activity. The results also indicated that different particle characteristics may activate different signalling pathways, since inhibition of endotoxin reduced release of TNF-alpha, IL-1beta and IL-8, whereas organic extraction only affected release of TNF-alpha and IL-8. Together, these data illustrate that a similar inflammatory response may be mediated by different particle characteristics and possibly through different signalling pathways. PMID- 18406507 TI - MAPK regulate p53-dependent cell death induced by benzo[a]pyrene: involvement of p53 phosphorylation and acetylation. AB - Benzo[a]pyrene (BaP) is a potentially genotoxic and cytotoxic environmental pollutant. Previous studies showed that exposure of HepG(2) cells to BaP causes necrotic cell death [Lin, T., Yang, M.S., 2007b. Cell death induced by benzo[a]pyrene in the HepG(2) cells is dependent on PARP-1 activation and NAD depletion. Toxicology 245, 147-153]. In the present study, the signaling pathways associated with this response was studied. BaP induced accumulation and activation of p53 in HepG(2) cells, which occurred as early as 12h after exposure. Activation of p53 was evidenced by its phosphorylation at serine 15 (Ser15) and acetylation at lysine 382 (Lys382). Chemical inhibition and siRNA mediated knockdown of p53 expression suppressed its phosphorylation as well as cell death. BaP also activated p38 MAPK and ERK, but not JNK, at 6h after exposure. SB203580 and PD98059, specific inhibitors of p38 MAPK and ERK, respectively, suppressed phosphorylation of p53 at Ser15, but the accumulation of p53 was only moderately reduced. Acetylation of p53 at Lys 382 was not affected by these inhibitors, suggesting that acetylation stabilizes p53 in response to DNA damage. SB203580 and PD98059 prevented downstream energy failure and BaP induced cell death. Similar results were obtained with siRNA against two isoforms of p38 MAPK, p38alpha and p38beta. Wortmannin, selective inhibitor of DNA-PK and ATM/ATR, abolished p53 phosphorylation, indicating an involvement of multiple pathways of p53 phosphorylation upon exposure to BaP. In summary, the current study demonstrated that both MAPK and p53 activation are required for BaP-induced necrotic cell death. The results also provide a novel model for studying the regulation between p53 and p38 MAPK in the progression of cellular necrosis. PMID- 18406508 TI - Maternal venous thrombosis. AB - Venous thrombosis remains a significant cause of maternal mortality and long-term morbidity, both in the developing and developed world. This review details the pathophysiology of venous thromboembolism in pregnancy and outlines the appropriate risk assessment, prophylaxis and management, which is required to minimise the burden of venous thrombosis on maternal health. PMID- 18406509 TI - Cervical fluid cytokines in pregnant women: Relation to vaginal wet mount findings and polymorphonuclear leukocyte counts. AB - OBJECTIVE: The purpose of the study was to measure and compare concentrations of IFN-gamma, IL-8, IL-10 and IL-12 in cervical fluids of pregnant women with normal and pathologic microscopic findings of vaginal wet mount. STUDY DESIGN: Vaginal samples were obtained from 100 pregnant women (22-32 weeks of gestation) without signs and symptoms of preterm labour (PTL), selected on the basis of the microscopic examination of vaginal wet mounts. Cytokine (IFN-gamma, IL-8, IL-10 and IL-12) concentrations were measured in cervical fluids by an enzyme-linked immunosorbent assay. RESULTS: The cytokines were found in cervical fluids of the majority of pregnant women, with IFN-gamma and IL-8 detected in all samples. Concentrations of IL-8 were significantly higher in pregnant women with pathologic findings on vaginal wet mount, including elevated polymorphonuclear (PMN) cell counts. CONCLUSION: A significant correlation between microscopic findings and cervical fluid IL-8 concentrations may make vaginal wet mount microscopy a useful tool for identification of patients with cervicovaginal inflammation, which is considered a risk factor for spontaneous preterm delivery. PMID- 18406510 TI - Fetal anterior abdominal wall thickness in diabetic pregnancy. AB - OBJECTIVE: The purpose of this study was to investigate whether third trimester fetal anterior abdominal wall (AAW) thickness in diabetic pregnancy reflects glycaemic control and predicts macrosomia. STUDY DESIGN: Prospective cohort study in a tertiary level maternity unit. One hundred and twenty-five diabetic mothers (71 pre-gestational and 54 gestational diabetics on insulin) underwent routine serial third trimester ultrasound examination with the additional measurement of AAW thickness. Pregnancy outcome was obtained. RESULTS: 335 fetal AAW measurements were recorded in diabetic pregnancy from 30 to 38 weeks gestation. Third trimester AAW was significantly higher in macrosomic babies (5.4+/-1.4 mm vs. 4.7+/-1.4 mm, p<0.05). ROC derived cut off for AAW in the prediction of macrosomia was 3.5 mm at 30 weeks, 4.5 mm at 33 weeks and 5.5 mm at 36 weeks gestation. Using either a raised AAW measurement or an AC>90th centile, the prediction of birth weight greater than the 90th centile was better (88%) than with AC alone (70%). This improvement in sensitivity held even at earlier gestations in the third trimester. CONCLUSION: Measurement of AAW in diabetic pregnancy may have a role in the prediction of macrosomia. PMID- 18406511 TI - Estimation of the effect of the degree of sewage treatment on the status of pollution along the coastline of the Mediterranean Sea using broad scale modelling. AB - A preliminary investigation was performed to estimate the effect of the degree of treatment in Sewage Treatment Plants (STPs) on the status of pollution along the coastlines of the Mediterranean Sea. Data from questionnaires and the literature were collected and processed (a) to identify 18 approximate 1D surface coastal currents, (b) to estimate their prevailing direction and average flow velocity and (c) to estimate the water pollution loads and identify their locations of discharge. Then, a simplified 1D water quality model was formulated and applied for the existing conditions and two hypothetical scenarios: (1) all coastal cities have STPs with secondary treatment and (2) all coastal cities have STPs with tertiary treatment to determine BOD(5), TN and TP concentrations in the 18 surface coastal currents. Calculated concentrations were compared and discussed. A main conclusion is that, to reduce pollution in the Mediterranean Sea measures should be adopted for upgrading the water quality of the rivers discharging into the Mediterranean Sea, along with the construction of STPs for all the coastal cities. PMID- 18406512 TI - The role of remote sensing in hydrological modelling of the Okavango Delta, Botswana. AB - A coupled surface water-groundwater model of the Okavango Delta has been built based on the United States Geological Survey software MODFLOW 2000 including the SFR2 package for stream-flow routing. It will provide a new tool for evaluating water management and climate change scenarios. The delta's size and limited accessibility make direct, on the ground data acquisition difficult. Remote sensing methods are the most promising source of acquiring spatially distributed data for both model input parameters and calibration. Topography, aquifer thickness, channel positions, evapotranspiration and precipitation data are all based on remote sensing. Simulated flooding patterns are compared to patterns derived from visible to thermal NOAA-AVHRR data and microwave radar ENVISAT-ASAR data. PMID- 18406513 TI - Comparison of two stream temperature models and evaluation of potential management alternatives for the Speed River, Southern Ontario. AB - Concerns over increased water temperature of the Speed River as it flows through the City of Guelph in Southern Ontario and an observed relationship between summer stream temperatures and low dissolved oxygen levels in the river prompted an investigation into potential stream temperature management practices. Two mechanistic stream temperature models, SNTEMP and CE-QUAL-W2, were applied to the Speed River in order to gauge the effectiveness of various stream temperature management options. Calibrated versions of both models performed well (0.2 degrees CTA transversion mutations. UVA-induced mutant frequency was measured in the guanine phosphoribosyl transferase (gpt) gene of Chinese hamster ovary cells (AS52) that were stably transfected to overexpress the hOGG1 protein, the human DNA repair glycosylase for 8-oxoG. This mutant frequency was compared with ultraviolet A induced mutant frequency in AS52 cells stably transfected with the same vector without the hOGG1 gene. The mutant frequency was significantly decreased in the hOGG1 overexpressing cells irradiated with 300 and 400 kJ/m2 ultraviolet A radiation, corresponding to 25% and 10% cell survival, respectively. The hOGG1 overexpressing cells repaired oxidative DNA lesions three times faster than the vector only cells as measured by a semi-automated alkaline elution assay using FPG enzyme, the bacterial OGG1 analogue, to cut DNA at oxidative base modifications. Thus, the lower mutation frequency in UVA-induced mutations of the hOGG1 overexpressing cells may be related to the increased repair of 8-oxoG. No GC-->TA mutations were detected in the UVA-irradiated hOGG1 overexpressing cells. The results suggest a link between the 8-oxoG lesion and UVA-induced mutagenesis. We propose that hOGG1 has a role in maintaining genomic stability in mammalian cells after oxidative stress. PMID- 18406517 TI - Safety assessment in plant layout design using indexing approach: implementing inherent safety perspective. Part 2-Domino Hazard Index and case study. AB - The design of layout plans requires adequate assessment tools for the quantification of safety performance. The general focus of the present work is to introduce an inherent safety perspective at different points of the layout design process. In particular, index approaches for safety assessment and decision making in the early stages of layout design are developed and discussed in this two-part contribution. Part 1 (accompanying paper) of the current work presents an integrated index approach for safety assessment of early plant layout. In the present paper (Part 2), an index for evaluation of the hazard related to the potential of domino effects is developed. The index considers the actual consequences of possible escalation scenarios and scores or ranks the subsequent accident propagation potential. The effects of inherent and passive protection measures are also assessed. The result is a rapid quantification of domino hazard potential that can provide substantial support for choices in the early stages of layout design. Additionally, a case study concerning selection among various layout options is presented and analyzed. The case study demonstrates the use and applicability of the indices developed in both parts of the current work and highlights the value of introducing inherent safety features early in layout design. PMID- 18406516 TI - Pro-inflammatory prostaglandins and progression of colorectal cancer. AB - Chronic inflammation is a risk factor for several gastrointestinal malignancies, including esophageal, gastric, hepatic, pancreatic and colorectal cancer. It has long been known that long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs) reduces the relative risk of developing colorectal cancer. NSAIDs exert their anti-inflammatory and anti-tumor effects primarily by inhibiting activity of cyclooxygenase (COX) enzymes. Cyclooxygenase enzymes catalyze the conversion of arachidonic acid into prostanoids, including prostaglandins (PGs) and thromboxanes (TXs). Emerging evidence demonstrates that prostaglandins play an important role in inflammation and cancer. In this review, we highlight recent breakthroughs in our understanding of the roles of the different prostaglandins in colorectal cancer (CRC) and inflammatory bowel disease (IBD). These findings may provide a rationale for the development of new anti-inflammatory therapeutic approaches to cancer prevention and/or treatment. PMID- 18406518 TI - An exploration of nitrate concentrations in groundwater aquifers of central-west region of Bangladesh. AB - Groundwater and river water samples were collected from the study area to investigate the spatial distribution of nitrate (NO(3)(-)) in the central-west region of Bangladesh. The shallow and deep groundwater nitrate concentrations ranged from <0.10 to 75.12 and <0.10 to 40.78 mg/L, respectively. Major river water NO(3)(-) concentrations were ranged from 0.98 to 2.32 mg/L with an average of 1.8 mg/L. The average Cl(-)/NO(3)(-) ratio (4.9) of major river water has been considered as reference point to delineate denitrification processes. The alluvial fan, alluvial, deltaic and coastal deposits shallow groundwater having C1(-)/NO(3)(-) values less than that of the average river water value (4.9), suggested denitrification processes within the aquifers. On the other hand, denitrification processes are insignificant in the Pleistocene terraces area aquifers related to relatively higher concentrations of nitrate. Iron pyrite has been found as insignificant effect on denitrification. PMID- 18406519 TI - Safety assessment in plant layout design using indexing approach: implementing inherent safety perspective. Part 1 - guideword applicability and method description. AB - Layout planning plays a key role in the inherent safety performance of process plants since this design feature controls the possibility of accidental chain events and the magnitude of possible consequences. A lack of suitable methods to promote the effective implementation of inherent safety in layout design calls for the development of new techniques and methods. In the present paper, a safety assessment approach suitable for layout design in the critical early phase is proposed. The concept of inherent safety is implemented within this safety assessment; the approach is based on an integrated assessment of inherent safety guideword applicability within the constraints typically present in layout design. Application of these guidewords is evaluated along with unit hazards and control devices to quantitatively map the safety performance of different layout options. Moreover, the economic aspects related to safety and inherent safety are evaluated by the method. Specific sub-indices are developed within the integrated safety assessment system to analyze and quantify the hazard related to domino effects. The proposed approach is quick in application, auditable and shares a common framework applicable in other phases of the design lifecycle (e.g. process design). The present work is divided in two parts: Part 1 (current paper) presents the application of inherent safety guidelines in layout design and the index method for safety assessment; Part 2 (accompanying paper) describes the domino hazard sub-index and demonstrates the proposed approach with a case study, thus evidencing the introduction of inherent safety features in layout design. PMID- 18406520 TI - Biosorption of total chromium from aqueous solution by red algae (Ceramium virgatum): equilibrium, kinetic and thermodynamic studies. AB - This study focused on the biosorption of total chromium onto red algae (Ceramium virgatum) biomass from aqueous solution. Experimental parameters affecting biosorption process such as pH, contact time, biomass dosage and temperature were studied. Langmuir, Freundlich and Dubinin-Radushkevich (D-R) models were applied to describe the biosorption isotherms. Langmuir model fitted the equilibrium data better than the Freundlich isotherm. The biosorption capacity of C. virgatum biomass for total chromium was found to be 26.5mg/g at pH 1.5 and 10g/L biomass dosage, 90min equilibrium time and 20 degrees C. From the D-R isotherm model, the mean free energy was calculated as 9.7kJ/mol, indicating that the biosorption of total chromium was taken place by chemisorption. The calculated thermodynamic parameters (DeltaG degrees , DeltaH degrees and DeltaS degrees ) showed that the biosorption of total chromium onto C. virgatum biomass was feasible, spontaneous and exothermic at 20-50 degrees C. Kinetic evaluation of experimental data showed that the biosorption processes of total chromium followed well pseudo-second order kinetics. PMID- 18406521 TI - Sorption of uranium(VI) from aqueous solutions by akaganeite. AB - Removal of U(VI) ions from aqueous solutions was investigated using synthetic akaganeite-type nanocrystals. Nanocrystals of iron oxyhydroxides were synthesized with two different methods and then compared their adsorption capacities. Akaganeite (beta-FeOOH) was synthesized in the laboratory by precipitation from aqueous solution of Fe(III) chloride and different precipitating agents. The relative importance of test parameters like solution pH, contact time, temperature and concentration of adsorbate on adsorption performance of akaganeite for U(VI) ion were studied. Typical adsorption isotherms (Langmuir, Freundlich, Dubinin-Raduskevich) were determined for the mechanism of sorption process. Also the thermodynamic constants (DeltaH degrees , DeltaS degrees and DeltaG degrees ) were calculated. The product materials were examined by powder X ray diffraction for crystalline phase identification and scanning electron microscope (SEM). PMID- 18406522 TI - Neuronal stimulation leading to upregulation of glutamate transporter-1 (GLT-1) in rat microglia in vitro. AB - As previously reported, activated microglia facilitate the expression of a glial cell-type glutamate transporter, glutamate transporter-1 (GLT-1; EAAT2), around injured motoneurons in axotomized rat facial nucleus. This phenomenon suggests that the motoneurons stimulate microglia to enhance the levels of GLT-1. In the present study, we investigated the effects of neuronal stimulus on the uptake of glutamate (Glu) by microglia and on the expression of GLT-1 protein in microglia in vitro. A 14C-Glu uptake experiment revealed that microglia enhance uptake of Glu by stimulation with neuronal conditioned medium (NCM). The NCM-stimulated uptake was significantly suppressed in the presence of dihydrokinate (a specific GLT-1 inhibitor), suggesting that GLT-1 is a major glutamate transporter for the uptake. Furthermore, immunoblotting analysis revealed that the amounts of GLT-1, but not another glial cell-type glutamate transporter glutamate-aspartate transporter (GLAST: EAAT1), increased significantly in microglia by treatment with NCM. Altogether, neuronal stimulus was found to promote the uptake of Glu in microglia, probably due to the increased levels of GLT-1. PMID- 18406523 TI - Enhanced folate binding of cultured fibroblasts from Alzheimer's disease patients. AB - We compared the levels of serum folate from Alzheimer's disease (AD) patients and from age-matched healthy subjects and used primary cultures of fibroblasts, obtained from the two groups, to assess possible differences in their ability to bind folate. The results show that the levels of circulating folate are significantly (p<0.01; n=30) lower in AD patients than in controls (4.91+/-2.44 and 7.56+/-2.5 ng/mL, respectively). Moreover, the folate binding of AD fibroblasts is significantly (p<0.01; n=8) higher (2-4-fold) with respect to controls. RT-PCR experiments suggest that the higher folate binding could be due to an enhanced expression in AD fibroblasts of folate receptor alpha. PMID- 18406524 TI - Spinal dis-inhibition in inflammatory pain. AB - Inflammatory diseases and neuropathic insults trigger signaling cascades, which frequently lead to intense and long-lasting pain syndromes in affected patients. Such pain syndromes are characterized not only by an increased sensitivity to painful stimuli (hyperalgesia), but also by a qualitative change in the sensory perception of other, tactile stimuli (allodynia) and the occurrence of spontaneous pain in the absence of any sensory input. Long-term potentiation (LTP)-like changes in synaptic transmission between nociceptive C-fibers and spino-periaqueductal grey projection neurons as well as a loss of inhibitory control by GABAergic and glycinergic spinal dorsal horn neurons have repeatedly been proposed as underlying principles. While considerable evidence supports a significant contribution of C-fiber LTP to hyperalgesia, such monosynaptic plasticity cannot explain the occurrence of allodynia and spontaneous pain. In this review, we focus on mechanisms of synaptic dis-inhibition in inflammatory pain and propose that pathologically heightened pain sensitivity can be reversed by restoring synaptic inhibition with drugs that target specific spinal GABAA receptor subtypes. PMID- 18406525 TI - Altered striatal vulnerability to 3-nitropropionic acid in rats due to sex hormone levels during late phase of brain development. AB - Systemic administration of 3-nitropropionic acid (3-NPA) leads to a shortage of cellular ATP and induces striatum-specific lesions that resemble Huntington's disease. Gender differences, in terms of vulnerability of striatum to 3-NPA, have been shown in male rats. The goal of the present study was to determine whether changes in sex hormone levels during the critical period of sexual differentiation (E17-P4) influence striatal vulnerability to 3-NPA. An androgen receptor antagonist, flutamide, or an aromatase-inhibitor, fadrozole hydrochloride, which block conversion of testosterone to estradiol, were administered to embryonic rats during E17-E20 or E18-E20, respectively, with subsequent 3-NPA (20mg/(kg day) for 2 days) treatment during adulthood (8-9 weeks old). Motor behavior and histological changes (IgG exudation due to blood-brain barrier dysfunction and glial fibrillary acidic protein immunoreactivity) were assessed. Treatment with flutamide significantly decreased the 3-NPA-induced motor behavior in male rats, while administration of fadrozole hydrochloride increased atypical motor behavior in female rats. IgG exudation, as well as decreased glial fibrillary acidic protein reactivity, was observed in animals with motor defects. Flutamide decreased testosterone levels in male rats, while fadrozole hydrochloride increased testosterone levels in female rats. These results suggest that prenatal modulation of sexual hormonal levels greatly influences vulnerability to 3-NPA during adulthood and directly correlates to serum testosterone levels. PMID- 18406526 TI - Circadian difference in firing rate of isolated rat suprachiasmatic nucleus neurons. AB - The suprachiasmatic nucleus (SCN) of the hypothalamus contains the primary circadian clock in mammals. Dissociated SCN neurons in long-term culture exhibit a circadian modulation of spontaneous electrical activity. To evaluate the presence of circadian differences in spontaneous activity of isolated SCN neurons without synaptic connections, dissociated rat SCN neurons were studied with on cell recording 3-4 days after preparation, before the formation of dendrites, axons and synapses. A day-night difference in spontaneous electrical firing rate was found in acutely dissociated SCN neurons. During the first subjective day, the average firing rate (0.87+/-0.12 Hz) was significantly higher than during the first subjective night (0.24+/-0.06 Hz), while the firing rate on the next day (0.68+/-0.11 Hz) was significantly higher than during the preceding night. These data suggest that populations of isolated SCN neurons with no synaptic interactions contain a functioning circadian clock, and are particularly amenable to biophysical experiments. PMID- 18406527 TI - The transthyretin gene is expressed in human and rodent dorsal root ganglia. AB - The transthyretin (TTR) gene is mainly expressed in the liver and choroid plexus of the brain. Most cases of familial amyloidotic polyneuropathy (FAP) are caused by TTR gene mutations, and characterized by amyloid deposition in the peripheral nervous system. We hypothesized that the TTR gene may be expressed in the peripheral nervous system. We analyzed TTR gene expression in several parts of the human, mouse and rat peripheral nervous systems using RT-PCR. To determine the sites of TTR synthesis in the dorsal root ganglia (DRG), mouse DRG were examined by in situ hybridization, laser capture microdissection and RT-PCR, and immunohistochemistry. TTR mRNA was detected in the DRG and cauda equina of humans and rodents by RT-PCR. TTR mRNA was not detected in the sural nerve, lumbar plexus or sympathetic ganglia in humans, or in the sciatic nerve in rodents. In mouse DRG, TTR mRNA was localized in the peripheral glial cells. No TTR-like immunoreactivity was observed in these tissues except for the perineurium. The TTR gene is probably expressed in the peripheral glial cells of the DRG. TTR synthesis in the DRG may be important for the involvement of the peripheral nervous system in FAP. PMID- 18406528 TI - Levetiracetam inhibits both ryanodine and IP3 receptor activated calcium induced calcium release in hippocampal neurons in culture. AB - Epilepsy affects approximately 1% of the population worldwide, and there is a pressing need to develop new anti-epileptic drugs (AEDs) and understand their mechanisms of action. Levetiracetam (LEV) is a novel AED and despite its increasingly widespread clinical use, its mechanism of action is as yet undetermined. Intracellular calcium ([Ca2+]i) regulation by both inositol 1,4,5 triphosphate receptors (IP3R) and ryanodine receptors (RyR) has been implicated in epileptogenesis and the maintenance of epilepsy. To this end, we investigated the effect of LEV on RyR and IP3R activated calcium-induced calcium release (CICR) in hippocampal neuronal cultures. RyR-mediated CICR was stimulated using the well-characterized RyR activator, caffeine. Caffeine (10mM) caused a significant increase in [Ca2+]i in hippocampal neurons. Treatment with LEV (33 microM) prior to stimulation of RyR-mediated CICR by caffeine led to a 61% decrease in the caffeine induced peak height of [Ca2+]i when compared to the control. Bradykinin stimulates IP3R-activated CICR-to test the effect of LEV on IP3R-mediated CICR, bradykinin (1 microM) was used to stimulate cells pre-treated with LEV (100 microM). The data showed that LEV caused a 74% decrease in IP3R mediated CICR compared to the control. In previous studies we have shown that altered Ca2+ homeostatic mechanisms play a role in seizure activity and the development of spontaneous recurrent epileptiform discharges (SREDs). Elevations in [Ca2+]i mediated by CICR systems have been associated with neurotoxicity, changes in neuronal plasticity, and the development of AE. Thus, the ability of LEV to modulate the two major CICR systems demonstrates an important molecular effect of this agent on a major second messenger system in neurons. PMID- 18406529 TI - Syntaxin 6 regulates nerve growth factor-dependent neurite outgrowth. AB - Neurite outgrowth is crucial for neural circuit formation. Intracellular membrane trafficking is involved in the cell surface expansion that is necessary for neurite outgrowth. It is known that syntaxin 6 is predominantly located in the Golgi region in undifferentiated PC12 cells and that it regulates trans-Golgi network trafficking and the secretory pathway via its coiled-coil domains. However, whether it also regulates neurite outgrowth remains unknown. In this paper, we found that syntaxin 6 was located both in the Golgi apparatus and the distal tips of the neurites of nerve growth factor (NGF)-treated PC12 cells. We also showed that the overexpression of the first coiled-coil domain of syntaxin 6 inhibited NGF-dependent neurite outgrowth. However, the coiled-coil domain disrupting mutant had little effect on neurite outgrowth. These results suggest that the first coiled-coil domain of syntaxin 6 plays a crucial role in NGF dependent neurite outgrowth. PMID- 18406530 TI - Escitalopram reduces increased hippocampal cytogenesis in a genetic rat depression model. AB - Hippocampal neurogenesis is potentially implicated in etiology of depression and as the final common mechanism underlying antidepressant treatments. However, decreased neurogenesis has not been demonstrated in depressed patients and, in animals, reduced cytogenesis was shown in healthy rats exposed to stressors, but, so far, not in models of depression. Here we report that the number of BrdU positive cells in hippocampus was (1) significantly higher in a rat model of depression, the Flinders Sensitive Line (FSL) compared to control FRL, (2) increased in both FSL and FRL following maternal separation, (3) reduced by escitalopram treatment in maternally separated animals to the level found in non separated animals. These results argue against the prevailing hypothesis that adult cytogenesis is reduced in depression and that the common mechanism underlying antidepressant treatments is to increase adult cytogenesis. The results also point to the importance of using a disease model and not healthy animals for testing effects of potential treatments for human depression and suggest other cellular mechanisms of action than those that had previously been proposed for escitalopram. PMID- 18406531 TI - AAV-mediated expression of Bcl-xL or XIAP fails to induce neuronal resistance against quinolinic acid-induced striatal lesioning. AB - Apoptotic mechanisms have been proposed to contribute to the selective loss of medium spiny striatal projection neurons in Huntington's disease (HD). This raises the question as to whether enhancing the expression of anti-apoptotic factors in vulnerable striatal projection neurons can reduce their susceptibility to neurotoxic processes occurring in the HD brain. In this study AAV 1/2 vectors encoding either the anti-apoptotic factor Bcl-xL or XIAP were used to transduce striatal neurons prior to an intrastriatal injection of the excitotoxic glutamate analogue quinolinic acid (QA). AAV 1/2 vector treated rats were observed in behavioural tests undertaken to assess whether anti-apoptotic factor expression provided amelioration of motor function impairment following unilateral QA induced striatal lesioning. AAV-XIAP treated rats displayed complete amelioration of an ipsilateral forelimb use bias relative to control animals. However, neither AAV-XIAP nor AAV-Bcl-xL treated rats demonstrated an improvement in sensorimotor neglect compared to control animals. Furthermore, we did not observe a significant reduction of QA-induced pathology in assessed neuronal populations of the basal ganglia. These results indicate that sole enhancement of XIAP or Bcl-xL is not sufficient to counteract QA-induced excitotoxic insult of striatal neurons. PMID- 18406532 TI - Novel carbon nanosheets as support for ultrahigh-resolution structural analysis of nanoparticles. AB - The resolution in transmission electron microscopy (TEM) has reached values as low as 0.08 nm. However, these values are not accessible for very small objects in the size range of a few nanometers or lower, as they have to be placed on some support, which contributes to the overall electron-scattering signal, thereby blurring the contrast. Here, we report on the use of nanosheets made from cross linked aromatic self-assembled monolayers as TEM sample supports. When transferred onto a copper grid, a single 1.6-nm-thick nanosheet can cover the grid and is free standing within the micron-sized openings. Despite its thinness, the sheet is stable under the impact of the electron beam. Micrographs taken from nanoclusters onto these nanosheets show highly increased contrast in comparison to the images taken from amorphous carbon supports. In scanning transmission electron microscopy with nanosheet support, a size analysis of sub-nanometer Au clusters was performed and single Au atoms were resolved. PMID- 18406533 TI - Identification of a LIM domain-containing gene in the Cyathostominae. AB - The Cyathostominae are a complex group of nematodes and are the primary parasitic pathogens of horses. Little is known of their basic biology. As part of an investigation into mechanisms involved in reactivation of mucosal larval stages, we identified a gene encoding a predicted LIM domain-containing protein (Cy-LIM 1). LIM domains are cysteine- and histidine-rich motifs that are thought to direct protein-protein interactions. Proteins that contain these domains have a wide range of functions including gene regulation, cell fate determination and cytoskeleton organization. The Cy-lim-1 mRNA was identified as an abundant transcript following differential display-arbitrary primed reverse transcriptase polymerase chain reaction amplification of RNA from faecal fourth stage larvae (FL4), which had been obtained from the diarrhoea of clinical cases of larval cyathostominosis. Detailed analysis showed that Cy-lim-1 was transcribed in FL4 and in other developmental stages; however there were differences in transcription of alternatively spliced variants amongst the stages. The predicted peptide sequence of Cy-lim-1 showed high identity to two LIM domain-containing proteins from Caenorhabditis elegans. RT-PCR analysis of these Cy-lim-1 homologues in C. elegans indicated that the two genes, which are described as separate entities in GenBank, are likely to compose a single gene of which alternative splice variants are transcribed. The LIM proteins from the cyathostomins and C. elegans were classified as LIM-only (LMO) proteins and, along with LMO proteins identified in sequence databases of other nematodes, comprise a group of LIM proteins distinct to those defined in other organisms. PMID- 18406534 TI - Sensitivity and specificity of various serological tests for the detection of Toxoplasma gondii infection in naturally infected sheep. AB - Comparative serological examination of 300 serum samples from sheep slaughtered in the main abattoir in Cairo, Egypt revealed a higher prevalence of toxoplasmosis (43.7%) with the modified agglutination test (MAT), followed by the enzyme linked immune-sorbant assay (ELISA) (41.7%) and the indirect fluorescent antibody test (IFAT) (37%), while the lowest prevalence was detected with the dye test (DT) (34%). When the data from the first three serological tests were compared with that of the DT test, which was used as a reference test for toxoplasmosis, MAT had the highest sensitivity (96%), followed by ELISA (90.1%) and IFAT, which demonstrated the lowest sensitivity (80.4%). Conversely, IFAT had the highest specificity (91.4%), followed by MAT (88.9%) and ELISA (85.9%). PMID- 18406535 TI - Garlic burns: case reports with an emphasis on associated and underlying pathology. PMID- 18406536 TI - The neuron-level phenomena underlying cognition and consciousness: synaptic activity and the action potential. AB - An unusual property of the neuron is its capability for cell-to-cell communication via synapses, known to be the neuron-level "protophenomenon" underlying the brain-level "real phenomenon" of cognition. The temporal synchronization of such synaptic activity is the leading candidate for explaining "cognitive binding" and therefore the unity of mind. An equally-unusual property of the neuron is the action potential, the means by which the neuron sends a signal down the axon. Although infrequently noted by researchers in relation to consciousness, signal propagation within the neuron entails the momentary permeability of the neuronal membrane, allowing a massive influx of charged ions into the cellular interior. Such openness to the extracellular world is arguably the protophenomenon of neuronal "sentience," literally, feeling the charge-state of the electrochemical environment. Sensitivity to the external pH is a common feature of all living cells, but is greatly amplified during the neuron's action potential. Synchronization of the action potentials of the same neurons that are involved in cognitive binding is the likely mechanism by which the sentience of individual neurons is coordinated into the brain-level phenomenon of subjective awareness. I conclude that a proper understanding of the permeability of the neuronal membrane during the action potential is as important for consciousness studies as is a proper understanding of synaptic transmission for the explication of the cognition made possible by neurons. PMID- 18406538 TI - Enhancement of photic shifts with the 5-HT1A mixed agonist/antagonist NAN-190: intra-suprachiasmatic nucleus pathway. AB - Chronic desynchronization between the mammalian circadian pacemaker and its external environment, such as that observed from shift work or jet lag, can lead to various long-term health consequences. The circadian clock can be reset by exposure to light, although the magnitude of such adjustments is modest. 5-HT modulates the effects of light, and 5-HT(1A) mixed agonist/antagonists, such as NAN-190, have been found to potentiate the phase resetting ability of light. The mechanism for this potentiation has yet to be uncovered, although it has been proposed that these drugs inhibit raphe output while simultaneously blocking post synaptic 5-HT(1A) receptors. The current study takes advantage of the heterogeneous network organization of the circadian clock to identify where in the circadian system NAN-190 exerts its influence. Retinorecipient cells in the ventrolateral suprachiasmatic nucleus (SCN) are activated by glutamate and release either gastrin-releasing peptide (GRP) or vasoactive intestinal polypeptide. Application of the glutamate agonist N-methyl-D-aspartic acid (NMDA) or either of these neuropeptides to the SCN mimics the effects of light. We hypothesized that NAN-190 would modify responses to treatments that activate the circadian system upstream, but not downstream, of where NAN-190 is acting. Hamsters were pretreated with NAN-190 or vehicle, followed by one of the neurochemicals 45 min later, during the early- and/or late-subjective night. NAN 190 potentiated NMDA-induced phase advances and delays as well as GRP-induced advances, but attenuated GRP-induced delays. NAN-190 did not potentiate NMDA induced Fos expression, however greater GRP-induced Fos expression was found within the dorsolateral region of the SCN. These data suggest that NAN-190 acts, in part, by modifying the responsiveness of retinorecipient cells in the circadian clock. An understanding of the neural events that underlie the potentiation of photic phase shifts by NAN-190 could guide the development of novel chronobiotics which could be used to treat a variety of sleep and circadian disorders. PMID- 18406537 TI - Estrogen induces rapid translocation of estrogen receptor beta, but not estrogen receptor alpha, to the neuronal plasma membrane. AB - Estrogen receptors can activate transcription in the nucleus, and activate rapid signal transduction cascades in the cytosol. Multiple reports identify estrogen receptors at the plasma membrane, while others document the dynamic responses of estrogen receptor to ligand binding. However, the function and identity of membrane estrogen receptors remain controversial. We have used confocal microscopy and cell fractionation on the murine hippocampus-derived HT22 cell line and rat primary cortical neurons transfected with estrogen receptor-green fluorescent protein constructs to address the membrane localization of these receptors. We observe translocation of estrogen receptor beta (beta) to the plasma membrane 5 min after exposure to 17beta-estradiol, whereas estrogen receptor alpha (alpha) localization remains unchanged. Membrane localization of estrogen receptor beta is transient, selective for 17beta-estradiol, and is not blocked by ICI182,780. Inhibition of the mitogen-activated protein kinase pathway does not block estrogen-mediated estrogen receptor beta membrane translocation, and in fact prolongs membrane localization. These data suggest that while both estrogen receptor alpha and estrogen receptor beta can be present at the neuronal membrane, their presence is differentially regulated. PMID- 18406539 TI - Soluble amyloid beta oligomers may contribute to apoptosis of retinal ganglion cells in glaucoma. AB - Glaucoma is one of the leading causes of visual impairment and blindness. It is characterized by excavation of optic nerve head and visual field loss. Even though the pathogenesis of glaucoma remains unclear, it is generally accepted that elevated intraocular pressure is the major risk factor. No matter what the specific initiators are, retinal ganglion cells are believed to die via apoptosis eventually. It is known that glaucoma correlates strongly with Alzheimer's disease and the two diseases share many similarities in pathogenic mechanisms. Recent studies have indicated that amyloid beta peptide, which is implicated in the progression of Alzheimer's disease, may be also responsible for retinal ganglion cells death in glaucoma. Amyloid beta exists in different forms, including monomers, oligomers and fibrils, and among these, as demonstrated by extensive evidences, soluble amyloid beta oligomers rather than insoluble amyloid beta fibrils induced apoptosis of neurons in Alzheimer's disease. Here we propose that soluble amyloid beta oligomers may play an important role in activation of apoptotic cascades in retinal ganglion cells in glaucoma. PMID- 18406540 TI - Keloid and hypertrophic scar: neurogenic inflammation hypotheses. AB - The mechanisms of fibroproliferation disease (FPD) of the skin, such as keloids and hypertrophic scars, are still unknown. Since mechanical stress appears to be an important factor for FPD generation, we have studied the intervening factors that connect mechanical stress with keloid and scar formation. Hence, we introduce our "neurogenic inflammation hypothesis" in this paper. Our hypothesis is as follows. Mechanical stress, including skin stretching, stimulates mechanosensitive nociceptors on sensory fibers in the skin. Stimulated fibers release neuropeptides, including SP and CGRP, and these peptides bind to the receptors SP-NK1R and CGRP-CGRP1R on various cells in the skin. Moreover, histamine release is upregulated by mast cells. Consequently, activated endothelial cells and vascular smooth muscle cells induce vasodilation and permeabilization of vessels. Cytokine production, including TGFbeta and NGF, is also stimulated by various cells. The neurogenic inflammation and upregulation of TGFbeta would activate fibroblasts through various signals. Interestingly, overexpressed NGF may induce the hyper-release of neuropeptides from sensory fibers, resulting in the accumulation of neuropeptides even in the absence of mechanical stress, once the malignant cycle has started. Moreover, individual differences in FPD generation may be based on differences in reactivity towards neuropeptides, NGF, and other neurotrophins. Hence, neuropeptide antagonists may be effective against FPD. While further experimental studies and clinical confirmation are needed, our hypothesis may provide new insights into the etiology and pathology of FPD of the skin, such as keloids and hypertrophic scars. PMID- 18406541 TI - Folate pathway genetic polymorphisms and susceptibility of central nervous system tumors in Thai children. AB - BACKGROUND: Folate is an important micronutrient molecule participating in DNA synthesis, methylation and repair mechanisms. Genetic polymorphisms in folate pathway related enzymes including methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C, methionine synthase (MTR) A2756G, thymidylate synthase (TS) 28 bp tandem repeat, and reduced folate carrier (RFC) G80A have been shown to be associated with increased susceptibility for several cancers. The aim of the present study was to evaluate whether single nucleotide polymorphisms in the genes encoding enzymes of the folate pathway predispose to any CNS tumors in Thai children. METHODS: In the present case-control study, we investigated these polymorphisms in genomic DNA from peripheral blood mononuclear cells in 73 Thai children with various types of central nervous system tumors and in 205 age and sex matched controls. RESULTS: Thirty-one out of 73 patients were diagnosed with glial tumors (astrocytoma, oigodendroglioma and ependymoma), 28 with embryonal CNS tumors (medulloblastoma, pinealoblastoma and primitive neuroectodermal tumor), 13 with germ cell tumors and 1 with meningioma. We found that the homozygous CC allele of MTHFR A1298C conferred an increased risk of embryonal CNS tumors (OR: 3.9; 95% CI: 1.3-11.4, p=0.02). CONCLUSION: Our findings thus suggest that folate metabolism may play a role in the pathogenesis of certain specific subtypes of pediatric brain tumor in Thai children, especially embryonal CNS tumors. PMID- 18406542 TI - [Breast metastasis: anatomoclinical study of six cases]. AB - INTRODUCTION: Breast metastases are rare. They represent 0.4 to 6% of all breast cancers. Our aim is to discuss the means of diagnosis and the clinicopathological features. PATIENTS AND METHODS: We report a retrospective survey of six cases of breast metastases diagnosed over a period of 11 years (1992-2003) in the laboratory of anatomy and pathological cytology of the university hospital of Sfax. The diagnosis was carried on a material of cytoponction in two cases, a biopsy in three cases, a surgery specimen in one case. Immunohistochemical study was performed in four cases. Clinical, therapeutic and evolutionary data were collected from the files of patients. RESULTS: Metastases to the breast constituted 0.43% of all breast cancers. The primary tumors understood a case of gingival-maxillary non-Hodgkin's lymphoma, a case of retroauricular melanoma, a case of soft tissue leiomyosarcoma, a case of uterine choriocarcinoma, a case of rectal neuroendocrine carcinoma and a case of gastric signet cell carcinoma. All patients were women, the middle age was 45.5 years. In three cases the metastases to the breast was concomitant to the diagnosis of the primitive tumour. Clinically it was a nodule in five cases, the size average was 2.3cm, and a subareolar thickening in one case. The mammary involvement was bilateral in two cases. The middle survival after the diagnosis was eight months. CONCLUSION: Metastases to the breast must be distinguished from primary breast cancers whose treatment and outcome are different. A confrontation of clinical and pathological data is recommended for an accurate diagnosis. Immunohistochemical study is of great interest particularly when the breast tumour is revealing the disease. PMID- 18406544 TI - [What do French trainees in obstetrics and gynecology wish as future professional activity in 2006? A national survey]. AB - OBJECTIVE: To survey French trainees in obstetrics and gynaecology on the type of short and long-term professional activity they wish to have following specialisation. MATERIAL AND METHODS: A questionnaire was sent to the 909 trainees in obstetrics and gynaecology during the academic year 2005-2006. RESULTS: Answer rate was 34.5%. Almost all trainees (96.5%) wish to have a two year hospital position before applying for consultant or beginning a private practice. Only 55.6% of these trainees think this option realistic. Overall, 78.7% of trainees wish to continue practicing obstetrics, 66.9% surgery, 41.4% ultrasound screening or prenatal diagnosis, 34.1% office gynaecology, 28% oncogynaecology and 23.6% assisted medical procreation. An exclusive public hospital position was desired by 51.6% of trainees, with 45.1% in academic hospitals and 54.9% in non academic hospitals. Exclusive private practice was desired by 8.9% of trainees and 47.7% wish a private practice with a part-time public hospital position. CONCLUSION: Actual trainees in obstetrics and gynaecology see their future practice as polyvalent, mostly in public hospitals and including for most at least obstetrics. PMID- 18406545 TI - The stay/switch model describes choice among magnitudes of reinforcers. AB - The stay/switch model is an alternative to the generalized matching law for describing choice in concurrent procedures. The purpose of the present experiment was to extend this model to choice among magnitudes of reinforcers. Rats were exposed to conditions in which the magnitude of reinforcers (number of food pellets) varied for staying at alternative 1, switching from alternative 1, staying at alternative 2 and switching from alternative 2. A changeover delay was not used. The results showed that the stay/switch model provided a good account of the data overall, and deviations from fits of the generalized matching law to response allocation data were in the direction predicted by the stay/switch model. In addition, comparisons among specific conditions suggested that varying the ratio of obtained reinforcers, as in the generalized matching law, was not necessary to change the response and time allocations. Other comparisons suggested that varying the ratio of obtained reinforcers was not sufficient to change response allocation. Taken together these results provide additional support for the stay/switch model of concurrent choice. PMID- 18406546 TI - Developmental and tissue-specific accumulation pattern for the Drosophila melanogaster TART ORF1 protein. AB - The TART, HeT-A, and TAHRE families of Drosophila non-LTR retrotransposons specifically retrotranspose to telomeres to maintain telomeric DNA. Recent evidence indicates that an RNA interference mechanism is likely to regulate TART, HeT-A, and TAHRE retrotransposition, but the developmental and tissue-specific expression of telomeric retrotransposon proteins has not previously been investigated. We have generated antisera against TART ORF1 protein (ORF1p) and used these antisera to examine the pattern of TART ORF1p expression in Drosophila melanogaster. We detected TART ORF1p throughout most of development and observed particularly high levels of protein in late larval and pupal stages. In late stage larvae, ORF1p accumulates in brain and imaginal discs tissues, rather than in terminally differentiated larval tissues. Accumulation of ORF1p in imaginal discs is intriguing, since TART antisense RNA has previously been detected in imaginal discs, and we discuss the implications of these findings for TART regulation. PMID- 18406547 TI - Strain-associated virulence factors of Streptococcus iniae in hybrid-striped bass. AB - Streptococcus iniae is a major fish pathogen producing invasive infections that result in economic losses in aquaculture. Development of in vitro models of S. iniae virulence may provide insight to the pathogenesis of infection in vivo. Three S. iniae strains (K288, 94-426, and 29178) were tested for virulence in a hybrid-striped bass (HSB) model using intraperitoneal injection. S. iniae strains K288 and 94-426 caused high levels of mortality in HSB (lethal dose 2x10(5)CFU) while strain 29178 was avirulent even upon IP challenge with 1000-fold higher inocula. In vitro assays were developed to test for the presence of characteristics previously associated with virulence in other species of pathogenic Streptococcus in animals and humans. In vitro differences relevant to virulence were not detected for beta-hemolysin activity, sensitivity to antimicrobial peptides, or adherence and invasion of epithelial cell layers. However, in whole-blood killing assays, the pathogenic strains were resistant to blood clearance, while 29178 was cleared (P<0.001) and more sensitive to complement (P<0.001). The avirulent strain 29178 was most efficiently phagocytosed and was most susceptible to intracellular killing (P<0.01) by the carp leukocyte cell line (CLC). When exposed to reactive oxygen species, strain 29178 was most susceptible. When the oxidative burst of CLC cells was inhibited, intracellular survival of 29178 was rescued fivefold, while no significant enhancement in survival of K288 or 94-426 was detected. Our results indicate that resistance to phagocytosis, oxidative killing, and associated phagocytic clearance is a significant factor in S. iniae virulence. PMID- 18406548 TI - Significant decreasing trend in human dietary exposure to PCDD/PCDFs and PCBs in Catalonia, Spain. AB - The concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/PCDFs), and 18 polychlorinated biphenyls (PCBs) were determined in samples of foodstuffs widely consumed by the population of Catalonia, Spain. The dietary intake of PCDD/PCDFs and dioxin-like (DL)-PCBs was subsequently estimated for the population of this Spanish region. These results were compared with those of a previous survey performed during 2000. For PCDD/PCDFs, the highest WHO-TEQ values corresponded to oils and fats (0.223 ng/kg), followed by fish and seafood (0.131 ng/kg) and dairy products (0.057 ng/kg), while the lowest levels were found in fruits (0.003 ng/kg), as well as in vegetables and milk (0.009 ng/kg). For DL PCBs the highest WHO-TEQ values corresponded to the groups of fish and seafood (0.761 ng/kg) followed by oils and fats (0.169 ng/kg), and dairy products (0.039 ng/kg), while the lowest values were observed in fruits (0.004 ng/kg), and vegetables (0.005 ng/kg) and tubers (0.006 ng/kg). The current dietary intakes of PCDD/PCDFs, DL-PCBs, and PCDD/PCDFs plus DL-PCBs were estimated to be 25.7, 52.4, and 78.1 pg WHO-TEQ/day vs. 95.4, 150.1, and 245.5 pg WHO-TEQ/day found in our previous survey. It means reductions of 73%, 65%, and 68%, for PCDD/PCDFs, DL PCBs, and PCDD/PCDFs plus DL-PCBs, respectively. The current estimated intake for an adult male, 1.12 pg WHO-TEQ/kg body weight per day, is lower than most intakes recently reported in a number of countries over the world. PMID- 18406549 TI - Hepatoprotective effects of Ganoderma lucidum peptides against D-galactosamine induced liver injury in mice. AB - Ganoderma lucidum (GL), a traditional Chinese medicinal mushroom, has been widely used for the treatment of chronic hepatopathy of various etiologies. The hepatoprotective activity of peptides from Ganoderma lucidum (GLP) was evaluated against d-galactosamine (d-GalN)-induced hepatic injury in mice. GLP was administered via gavage daily for 2 weeks at doses of 60, 120 and 180 mg/kg, respectively. Control groups were given the same amount of physiological saline synchronously. Then the mice from d-GalN control and GLP-treated groups were treated with d-GalN (750 mg/kg) suspended in normal saline by intraperitoneal injection. d-GalN-induced hepatic damage was manifested by a significant increase in the activities of marker enzymes (AST, ALT) in serum and MDA level in liver (P<0.01), and by a significant decrease in activity of SOD and GSH level in liver (P<0.01). Pretreatment of mice with GLP reversed these altered parameters to normal values. The biochemical results were supplemented by histopathological examination of liver sections. The best hepatoprotective effects of GLP were observed after treatment with the dose of 180 mg/kg as it was evidenced from biochemical parameters and liver histopathological characters which were similar to those of normal control group. Results of this study revealed that GLP could afford a significant protection in the alleviation of d-GalN-induced hepatocellular injury. PMID- 18406550 TI - Accuracy and reliability of breath alcohol testing by handheld electrochemical analysers. AB - Usefulness of portable, handheld breath analysers equipped with electrochemical sensor was assessed. Breath- and blood-alcohol concentrations in drunken drivers were taken from 370 expert opinions elaborated at the Institute of Forensic Research between January 1st 2002 and February 28th 2007. The results of second and subsequent measurements were re-calculated using mean elimination rates. The readings of portable instruments were in very good agreement with the results of confirmatory analyses performed by stationary devices (r=0.978, p<0.001, y=0.969x 0.0002). The correlation with the results of blood analysis was weaker (r=0.940, p<0.001, y=1.722x+0.214), but comparable with the correlation between the readings of stationary devices and the results of blood analyses (r=0.936, p<0.001, y=1.790x+0.091). The readings of portable and stationary breath analysers were also compared by the Bland-Altman plots. The differences in results were independent of alcohol concentration (absolute difference (mg/L): r=0.054, p>0.1, y=0.011x+0.013; relative difference (%): r=0.020, p>0.1, y=0.90x+2.36). PMID- 18406551 TI - A changing spectrum: mucosal damage and the clinical picture of adult celiac disease. PMID- 18406552 TI - [Dysmetabolic iron overload syndrome]. PMID- 18406553 TI - Modulation of longevity and diapause by redox regulation mechanisms under the insulin-like signaling control in Caenorhabditis elegans. AB - In Caenorhabditis elegans, the downregulation of insulin-like signaling induces lifespan extension (Age) and the constitutive formation of dauer larvae (Daf-c). This also causes resistance to oxidative stress (Oxr) and other stress stimuli and enhances the expression of many stress-defense-related enzymes such as Mn superoxide dismutase (SOD) that functions to remove reactive oxygen species in mitochondria. To elucidate the roles of the two isoforms of MnSOD, SOD-2 and SOD 3, in the Age, Daf-c and Oxr phenotypes, we investigated the effects of a gene knockout of MnSODs on them in the daf-2 (insulin-like receptor) mutants that lower insulin-like signaling. In our current report, we demonstrate that double deletions of two MnSOD genes induce oxidative-stress sensitivity and thus ablate Oxr, but do not abolish Age in the daf-2 mutant background. This indicates that Oxr is not the underlying cause of Age and that oxidative stress is not necessarily a limiting factor for longevity. Interestingly, deletions in the sod 2 and sod-3 genes suppressed and stimulated, respectively, both Age and Daf-c. In addition, the sod-2/sod-3 double deletions stimulated these phenotypes in a similar manner to the sod-3 deletion, suggesting that the regulatory pathway consists of two MnSOD isoforms. Furthermore, hyperoxic and hypoxic conditions affected Daf-c in the MnSOD-deleted daf-2 mutants. We thus conclude that the MnSOD systems in C. elegans fine-tune the insulin-like-signaling based regulation of both longevity and dauer formation by acting not as antioxidants but as physiological-redox-signaling modulators. PMID- 18406554 TI - (1)H MR spectroscopy of human brain tumours: a practical approach. AB - Magnetic resonance spectroscopy (MRS) is proposed in addition to magnetic resonance imaging (MRI) to help in the characterization of brain tumours by detecting metabolic alterations that may be indicative of the tumour class. MRS can be routinely performed on clinical magnets, within a reasonable acquisition time and if performed under adequate conditions, MRS is reproducible and thus can be used for longitudinal follow-up of treatment. MRS can also be performed in clinical practice to guide the neurosurgeon into the most aggressive part of the lesions or to avoid unnecessary surgery, which may furthermore decrease the risk of surgical morbidity. PMID- 18406555 TI - Radiological detectability of minute hepatic venous invasion in hepatocellular carcinoma. AB - OBJECTIVE: To determine if minute hepatic venous invasion in hepatocellular carcinoma (HCC) can be diagnosed radiologically. MATERIALS AND METHODS: CT hepatic arteriography (CTHA) and CT arterioportography (CTAP) of 95 cases with HCCs were examined. Histopathology after surgery has been the gold standard in all patients. Based on the presence of microscopic portal venous invasion (MPVI) and microscopic hepatic venous invasion (MHVI), the cases were classified into four groups as follows: Group vp0vv0, negative MPVI and MHVI; Group vp1vv0, positive MPVI and negative MHVI; Group vp0vv1, negative MPVI and positive MHVI; Group vp1vv1, positive MPVI and MHVI. An area showing low attenuation on CTAP and high attenuation on CTHA around the tumor was defined as an area of peritumoral hemodynamic change (APTHC). The shape and size of APTHC were compared between Groups vp0vv1 and vp0vv0 or between Groups vp1vv1 and vp1vv0. The ratio of APTHC volume to tumor volume (RAT) was employed as an indicator of APTHC size. Each comparison was also made independently when tumor diameter was limited to either less than 3 cm or 3 cm or more. RESULTS: Three types of APTHC were identified: wedge-shaped, belt-shaped or irregular, and linear. No significant difference in the frequency of each type of APTHC was observed between Groups vp0vv1 and vp0vv0 or between Groups vp1vv1 and vp1vv0. There was no significant difference in RAT between Groups vp0vv1 and vp0vv0 or between Groups vp1vv1 and vp1vv0, unrelated to tumor size. CONCLUSIONS: The presence of minute hepatic venous invasion in HCC is difficult to determine even on combined CTHA and CTAP. PMID- 18406556 TI - MRI of shoulder instability. AB - The most unstable joint in the body, the glenohumeral joint is subject to many insults including microinstability, subluxation and dislocation. During the last two decades, MRI has allowed for direct visualization of many of the lesions related to instability, aiding in diagnosis as well as therapeutic planning and follow-up. This article reviews the use of MRI for shoulder instability and describes the different types of lesions associated with this disorder. Topics include technical considerations, the use of MR arthrography, normal anatomy and variants, labral and glenohumeral ligament pathology, and osseous lesions related to instability. PMID- 18406557 TI - Miscellaneous conditions of the shoulder: anatomical, clinical, and pictorial review emphasizing potential pitfalls in imaging diagnosis. AB - The purpose of this article is to review the key imaging findings in major categories of pathology affecting the shoulder joint including hydroxyapatite deposition disease, rotator cuff interval pathology, acromioclavicular joint pathology, glenohumeral osteoarthrosis, and synovial inflammatory processes, with specific emphasis on findings that have associated pitfalls in imaging diagnosis. The pathophysiology and clinical manifestations of the above mentioned categories of pathology will be reviewed, followed in each section by a detailed pictorial review of the key imaging findings in each category including plain film, computed tomography, and magnetic resonance imaging findings as applicable. Imaging challenges that relate to both diagnosis and characterization will be addressed with each type of pathology. The goal is that after reading this article, the reader will be able to recognize the key imaging findings in major categories of pathology affecting the shoulder joint and will become familiar with the potential pitfalls in their imaging diagnosis. PMID- 18406558 TI - "Bubble-like" lung metastases in osteosarcoma patients. AB - PURPOSE: Cavitation of pulmonary metastases have been reported by several authors either as a spontaneous phenomenon or as a consequence of chemotherapy. We present two cases, with this type of image in follow-up, and 20-45 months after the end of treatment. This was the first sign of pulmonary metastases. RESULTS: Two patients with osteogenic sarcoma developed radiological evidence of pulmonary "bubble-like" cavitation several years following completion of chemotherapy. In one patient the "bubble-like" cavitation transformed into a solid nodule. Both patients had surgical resections of all pulmonary lesions, and histology confirmed presence of viable osteosarcoma metastases. CONCLUSION: The two cases suggest that onset of "bubble-like" cavitation in lung parenchyma of osteosarcoma patients may be the first sign of pulmonary metastases. PMID- 18406559 TI - The cloned 1-aminocyclopropane-1-carboxylate (ACC) deaminase gene from Sinorhizobium sp. strain BL3 in Rhizobium sp. strain TAL1145 promotes nodulation and growth of Leucaena leucocephala. AB - The objective of this study was to determine the role of 1-aminocyclopropane-1 carboxylate (ACC) deaminase of symbionts in nodulation and growth of Leucaena leucocephala. The acdS genes encoding ACC deaminase were cloned from Rhizobium sp. strain TAL1145 and Sinorhizobium sp. BL3 in multicopy plasmids, and transferred to TAL1145. The BL3-acdS gene greatly enhanced ACC deaminase activity in TAL1145 compared to the native acdS gene. The transconjugants of TAL1145 containing the native or BL3 acdS gene could grow in minimal media containing 1.5mM ACC, whereas BL3 could tolerate up to 3mM ACC. The TAL1145 acdS gene was inducible by mimosine and not by ACC, while the BL3 acdS gene was highly inducible by ACC and not by mimosine. The transconjugants of TAL1145 containing the native- and BL3-acdS genes formed nodules with greater number and sizes, and produced higher root mass on L. leucocephala than by TAL1145. This study shows that the introduction of multiple copies of the acdS gene increased ACC deaminase activities of TAL1145 and enhanced its symbiotic efficiency on L. leucocephala. PMID- 18406560 TI - Regulation of RALDH-1, RALDH-3 and CYP26A1 by transcription factors cVax/Vax2 and Tbx5 in the embryonic chick retina. AB - In vertebrate development, polarity of the dorsal-ventral eye axis is determined by the spatially restricted expression of transcription factors cVax/Vax2 in the ventral retina and Tbx5 in the dorsal retina. In addition, an early gradient of retinoic acid (RA) has been suggested to be involved in this process. Using retroviral transfection of the early eye anlage of chick embryos we investigated whether cVax/Vax2 and Tbx5 regulate the expression of enzymes that determine the retinal distribution of RA. Transfection of cVax/Vax2 completely abolished the spatial expression pattern of the RA synthesizing enzyme RALDH-1, of the RA degrading enzyme Cyp26A1 and caused an ectopic expression of RALDH-3 in the dorsal retina. Transfection of Tbx5 did not significantly alter the distribution of RALDH-1 and RALDH-3 but caused additional patches of Cyp26 outside of its normal domain of expression. The distribution of RA was experimentally changed either by ectopic expression of Cyp26A1, or by applications of RA. These manipulations did not affect the distribution of cVax/Vax2 or interfere with development of the retinotectal projection from dorsal retinal ganglion cells. The results show that in the chick retina the compartmentalized expression of RA metabolizing enzymes requires the spatially restricted expression of the transcription factors cVax/Vax2 and Tbx5. On the other hand, RA may not be decisive for the correct retinal projection toward the optic tectum. PMID- 18406561 TI - A pilot intervention for pregnant women in Sichuan, China on passive smoking. AB - OBJECTIVE: The study aimed to reduce women's exposure to passive smoking, which has been linked to increased risk of cancer and other diseases. By exploring the knowledge, attitudes and behaviors of pregnant women with regard to secondhand smoke in the home environment, a model for intervention during the pregnancy was designed with the help of the intended beneficiaries and was piloted. METHODS: The study had two phases. Phase 1 included focus group discussions and an iterative process to design an intervention. Phase 2 was the intervention itself, which included a series of motivational and patient communication activities, a resource booklet, clinician counseling, telephone hotline and regular telephone counseling. Pre- and post-intervention questionnaires were used to measure results. RESULTS: Post-intervention questionnaires showed a significant increase in knowledge, a change in attitudes towards stronger disapproval and an increased likelihood of taking assertive action when exposed to secondhand smoke in the family. PRACTICE IMPLICATIONS: The results of this study point to the fact that there is a need to give emphasis to passive smoking in the home environment and that the women's pregnancy is an effective conduit to increasing knowledge and bringing about change. The intervention model can become a part of the standard protocol for the care of pregnant women in hospital settings. PMID- 18406563 TI - Measurement and analysis of ultimate mechanical properties, stress-strain curve fit, and elastic modulus formula of human abdominal aortic aneurysm and nonaneurysmal abdominal aorta. AB - OBJECTIVE: The maximal strain, stress, elastic modulus, and stress-strain curve fitting of abdominal aortic aneurysms (AAA) and bidirectional nonaneurysmal abdominal aorta (NAA) were measured and analyzed to obtain the ultimate mechanical properties, the more approximate stress-strain curve-fitting, and the elastic modulus formula of AAA and NAA. METHODS: Fourteen human AAA samples were harvested from patients undergoing elective aneurysm repair. Twelve NAA samples comprised of six longitudinal-circumferential pairs of NAA from six cadaveric organ donors were used as controls. Samples were mounted on a tensile-testing machine and force was applied until breakage occurred. The maximal strain, stress, and elastic modulus were calculated and a stress-strain curve was plotted for each sample. Exponential and second-order polynomial curves were used to fit the stress-strain curve, and the means were estimated by comparing the R2 (coefficient of determination that represents the strength of a curve fitting). Coefficients of elastic modulus were calculated and analyzed, and the incremental tendency of each modulus was evaluated by comparing the difference of coefficients. RESULTS: There was no significant difference in maximal stress among AAA, circumferential aortic aneurysms (CAA), and longitudinal aortic aneurysms (LAA). However, AAA maximal strain was significantly less (P < .01) than that of bidirectional NAA. AAA maximal elastic modulus was significantly greater than that of CAA and LAA (P < .01 and .05, respectively). R2 of AAA for second-order polynomial curve was significantly greater (P < .05) than that for the exponential curve. For the elastic modulus formula from the second-order polynomial curve, E = 2ax + b, the average value of a for the AAA was significantly greater (P < .01) than that for the bidirectional NAA, but there was no significant difference (P > .05) among the three groups for the average value of b. CONCLUSIONS: Tensile test measurements can successfully analyze ultimate mechanical properties of AAA and NAA. AAA is stiffer and less distensible than NAA under the same maximal stress. Second-order polynomial curve fitting provides a more approximate description for AAA stress-strain curve than exponential curve fitting does. Formula variables a of the elastic modulus formula from second-order polynomial curve fitting can determine the incremental tendency of the elastic modulus, while b has negligible effect on the incremental tendency of the elastic modulus. PMID- 18406562 TI - How much time does it take to prescribe a new medication? AB - OBJECTIVE: To measure the length of time spent discussing all aspects of new prescriptions and guideline-recommended aspects of counseling, and to evaluate factors associated with duration of discussion. METHODS: We analyzed tape recordings in which 181 patients received 234 new medication prescriptions from 16 family physicians, 18 internists, and 11 cardiologists in 2 healthcare systems in Sacramento, California between January and November 1999. RESULTS: Of the mean total visit time of 15.9min (S.D.=434s), a mean of 26s (S.D.=28s) was allocated to guideline-recommended components and a mean of 23s (S.D.=25s) was allocated to discussion of all other aspects of new prescription medications. The majority of time spent discussing individual new prescriptions was dedicated to: medication purpose or justification, directions and duration of use, and side effects. On average, more complete discussion of these components was associated with more time. More time was spent talking about guideline-recommended information if patients were in better health, if there was a third party in the room, and if the medication belonged to a psychiatric, compared to an ear, nose, throat medication class. Less time was spent discussing over-the-counter (OTC) medications and those prescribed to patients with a previous visit to the physician. CONCLUSION: Higher quality information transmission between physicians and patients about new medications requires more time, and may be difficult to achieve in short office visits. PRACTICE IMPLICATIONS: Time-compressed office visits may need to be redesigned to promote improved provider-patient communication about new medications. PMID- 18406564 TI - [Neurotoxins from snake venom]. AB - Many snakes are able to quickly immobilize prey, thanks to their venom neurotoxins. Most of these snakes belong to families Elapidae or Hydrophidae but neurotoxins were also isolated from families Viperidae and Colubridae. Ophidian neurotoxins can be classified into several categories: neurotoxins which inhibit synaptic transmission (postsynaptic and presynaptic neurotoxins) and neurotoxins which facilitate it excessively (dendrotoxin and fasciculin). Their toxicity is dose-dependent, and venom effects are extremely fast. The clinical feature is a potentially fatal neurological syndrome, the so called cobraic syndrome. Because death by respiratory arrest may occur quickly with cobraic syndrome, immunotherapy is a true emergency, because toxins irreversible fixing makes immunotherapy effect uncertain after a few hours passed. PMID- 18406565 TI - Inhibition activity of sulfated polysaccharide of Sepiella maindroni ink on matrix metalloproteinase (MMP)-2. AB - SIP-SII is the sulfated S. maindroni ink polysaccharide (SIP) isolated from cuttlefish Sepiella maindroni. SIP-SII weakly inhibited tumor cell growth without cytotoxicity in vitro assay. Herein, we examined the effects of SIP-SII on the expression of matrix metalloproteinase MMP-2 and MMP-9 as well as tumor cell invasion and migration. SIP-SII (0.8-500 microg/ml) significantly decreased the expression of MMP-2 activity in human ovarian carcinoma cells SKOV3 as evidenced by the gelatin zymography analysis. No significant decrease of MMP-9 was detected in the cell line after SIP-SII treatment. The expression of MMP-2 was also evaluated using Western blot analysis. The results showed that SIP-SII inhibited the expression of MMP-2 in SKOV3 and human umbilical vein vascular endothelial cells ECV304 after 24 h incubation. Furthermore, the activity of invasion and migration of SKOV3 and ECV304 cells were measured. SIP-SII displayed an inhibitory effect on the penetration of SKOV3 cells through Matrigel-coated membrane in transwell chamber. A significant inhibition of ECV304 cell migration was observed in the presence of SIP-SII. These results suggest that SIP-SII might suppress invasion and migration of carcinoma cells via inhibition of MMP-2 proteolytic activity. PMID- 18406566 TI - Priming effect of homocysteine on inducible vascular cell adhesion molecule-1 expression in endothelial cells. AB - Hyperhomocysteinemia is an independent risk factor for the development of atherosclerosis, as well as for arterial and venous thrombosis. However, the mechanisms through which elevated circulating levels of homocysteine cause vascular injury and promote thrombosis remain unclear. Here, we tested the hypothesis that homocysteine (Hcy) sensitizes endothelial cells to the effect of inflammatory mediators. Human umbilical vein endothelial cells (HUVEC) were incubated with Hcy 1.0 mM for varying time points, and then treated in the absence or presence of 1.5 U/ml thrombin or 10 mg/ml lipopolysaccharide (LPS). Hcy alone had no effect on the expression of vascular cell adhesion molecule (VCAM)-1. However, Hcy enhanced thrombin- and LPS-mediated induction of VCAM-1 mRNA and protein levels. Consistent with these results, pretreatment of HUVEC with Hcy resulted in a two-fold increase in LSP-mediated induction of leukocyte adhesion. The latter effect was significantly inhibited by anti-VCAM-1 antibodies. Together, these findings suggest that Hcy sensitizes HUVEC to the effect of inflammatory mediators thrombin and LPS, at least in part through VCAM 1 expression and function. PMID- 18406567 TI - Effects of fibrates, anti-inflammatory drugs and antidepressants in the fish hepatoma cell line PLHC-1: cytotoxicity and interactions with cytochrome P450 1A. AB - Effects of 11 pharmaceuticals belonging to three therapeutic classes (lipid regulators from the fibrate group, non-steroidal anti-inflammatory drugs and anti depressives from the selective serotonin reuptake inhibitors group) were assessed in the fish hepatoma cell line (PLHC-1) by looking at cytotoxicity and interactions with cytochrome P450 1A (CYP1A) function. Among the tested pharmaceuticals, fluoxetine and paroxetine exerted cytotoxic effects, cell viability decreased to 52% and 6% after 24 h of exposure to 20 microM fluoxetine and paroxetine, respectively. The cytotoxicity of both compounds was modulated by cytochrome P450 inhibitors and was dramatically reduced when culture medium was supplemented with reduced glutathione and vitamin E succinate. Additionally, exposure of PLHC-1 cells to some pharmaceuticals led to an early and transient induction of ethoxyresorufin O-deethylase (EROD) activity: bezafibrate and antidepressants induced EROD activity at a concentration of 1 microM whereas clofibrate, ibuprofen and naproxen acted as inducers at a higher concentration (10 microM). These effects might be of toxicological concern since alterations of CYP1A may affect xenobiotic metabolism and toxicity. PMID- 18406568 TI - Inhibition of peroxide-induced radical generation by plant polyphenols in C6 astroglioma cells. AB - Plant polyphenols like flavonoids and hydroxystilbens have been found to possess radical scavenging/antioxidative activity, especially when studied in cell-free systems. A positive effect in such assays, however, does not necessarily indicate a protective activity against deleterious effects of oxidative stress in intact cells. In fact it has been shown that polyphenols can act as anti-oxidants as well as pro-oxidants. The aim of the present study was to investigate whether and with what potency selected polyphenols are able to inhibit cellular radical generation in C6 cells and whether they can induce oxidative stress themselves. Cumene hydroperoxide (CHP) was used as a model to induce radical generation which was measured by means of a fluorometric 2',7'-dichlorodihydro-fluorescein assay. CHP-induced, time and concentration dependent, a manifold increase of DCF fluorescence indicating intracellular radical generation. This process was inhibited by all the flavonoids and the hydroxystilben resveratrol, at low micromolar concentrations. The most potent compounds, luteolin and galangin, already at concentrations of 5 to 10 microM nearly completely abolished the radical generation in the presence of 500 microM CHP. The following ranking of anti-oxidative potency was obtained: luteolingalangin>kaempferol>quercetin>resveratrolgenisteintaxifolin. This ranking is completely different from that obtained by means of a trolox equivalent antioxidant capacity (TEAC) assay in a cell-free system, thus putting the biological relevance of the latter in question. Remarkably, one compound induced oxidative stress itself, namely genistein. This flavonoid inhibited the cellular radical generation in the presence of CHP while it significantly enhanced it in the absence of the peroxide. PMID- 18406569 TI - Concordance between telephone survey classification and face-to-face structured clinical interview in the diagnosis of generalized anxiety disorder in Hong Kong. AB - Telephone surveys of estimating mental disorders have been found to generate comparable findings to large-scale community surveys but the concordance between telephone instruments and clinical interviews is rarely examined. In this study, 100 Chinese respondents who had taken part in a telephone-based population survey of generalized anxiety disorder (GAD) in Hong Kong were administered the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I) by clinical interviewers. The discriminability and predictive indicators of the telephone survey instrument were assessed using receiver operating characteristic analysis. Results showed that the telephone survey instrument identified individuals with a positive SCID diagnosis of GAD better than those without. Although its individual questions performed well in identifying the endorsement of the corresponding core SCID criterion of GAD, further studies are needed to find out the optimal combination of questions in the telephone instrument for identifying GAD in community surveys. PMID- 18406570 TI - Multiplex RT-PCR assay for simultaneous detection of six viruses of penaeid shrimp. AB - In the present study, multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for simultaneously detection of six major shrimp viruses including yellow-head virus (YHV), white spot syndrome virus (WSSV), Taura syndrome virus (TSV), hepatopancreatic parvovirus (HPV), infectious hypodermal and hematopoietic necrosis virus (IHHNV) and monodon baculovirus (MBV). The six primer sets could amplify viral nucleic acids resulting in PCR products with different sizes. They were highly specific and did not cross-hybridize with other viral or shrimp nucleic acids. The sensitivity of the multiplex RT-PCR was 0.15pg for IHHNV, 0.15pg for TSV, 1.00pg for HPV, 1.5pg for MBV, 5.00pg for WSSV and 10.00pg for YHV. In the field application, 42 samples including whole tissue of post-larvae and hepatopancreas of Penaeus monodon collected from ponds in the central and southern parts of Thailand during 2002-2005 were examined by multiplex RT-PCR. The results revealed that a single infection was dominant and WSSV was the highest prevalence at that time. Dual infection was found in one sample. This developed multiplex RT-PCR will be useful for simultaneous detection of six major viruses of penaeid shrimp and benefit to shrimp cultured industry. PMID- 18406571 TI - Regionally selective changes in neurotransmitter receptors in the brain of the 5 HT1B knockout mouse. AB - The serotonin1B receptor knockout (5-HT1B KO) mouse is a valuable animal model of addiction to psychostimulants. We previously found selective increases in dopamine (DA) turnover in the nucleus accumbens of these mice, in addition to several changes in their central serotonin system. Here, we searched for further DA adaptations by measuring D1 and D2 receptor as well DA plasma membrane transporter (DAT) sites by ligand binding autoradiography, and G-protein coupling to D1 and D2 receptors by [35S]GTP gamma S autoradiography. Except for a slight increase in the lateral septum, D1 receptor binding did not differ from wild-type in twenty-one other neocortical, limbic or basal ganglia regions examined in the KO. Nor were there changes in D1 agonist-stimulated G-protein coupling in any of these regions, including the lateral septum. Increases in D2 binding sites, presumably involving GABAergic projection neurons, were measured in the nucleus accumbens, olfactory tubercle and ventral tegmental area of the 5-HT1B KO. However, no activation of the efficacy of D2 receptor coupling to G-protein could be measured in these and other brain regions. Binding to DAT was unchanged throughout brain. Because of their implication in cocaine addiction, the functionality of mu-opioid and GABAB receptors was also assessed by [35S]GTP gamma S autoradiography. 5-HT1B KO showed selective decreases in G-protein coupling to mu-opioid receptors in the paraventricular thalamic nucleus, and to GABAB receptors in the basolateral nucleus of amygdala. It is likely that these latter changes underlie some aspects of the addictive behavior of the 5-HT1B KO mouse. PMID- 18406572 TI - Regression equations for calculation of z scores of cardiac structures in a large cohort of healthy infants, children, and adolescents: an echocardiographic study. AB - BACKGROUND: Decision making in the care of pediatric patients with congenital and acquired heart disease remains reliant on detailed measurements of cardiac structures using 2-dimensional echocardiography. Calculated z scores are often used to normalize these measurements to the patient's body size. Existing normal data in the literature are limited by small sample size, small numbers of measured cardiac structures, and inadequate data for the calculation of z scores. Accordingly, we sought to develop normative data in a large pediatric cohort using modern echocardiographic equipment from which z scores could be calculated. METHODS: Two-dimensional and M-mode echocardiography was performed in 782 patients ranging in age from 1 day to 18 years. Measurements were made of 21 individual cardiac structures. Regression equations were derived to relate the size of the various cardiac structures to body surface area. Data are presented graphically, and regression equations are derived relating cardiac dimension to body surface area. CONCLUSION: The presented data will allow the calculation of z scores for echocardiographically measured cardiac structures. This information will be valuable for clinicians caring for infants and children with known or suspected cardiac disease. PMID- 18406573 TI - Early regional myocardial dysfunction in young patients with Duchenne muscular dystrophy. AB - BACKGROUND: In young patients (aged <12 years) with Duchenne muscular dystrophy (DMD), cardiac systolic function is generally described to be within the normal range. Recent studies have suggested the presence of subclinical dysfunction in these young patients using cardiac magnetic resonance imaging, tissue Doppler measurements, and myocardial velocity gradients. The aim of this study was to further assess regional myocardial function in a young group of patients with DMD using myocardial velocity and deformation imaging. METHODS: Thirty-two patients with DMD (mean age, 7.9 years; range, 3-12 years) and 29 age-matched normal controls were studied with echocardiography. Standard echocardiographic measurements of left ventricular (LV) systolic and diastolic function were performed. Myocardial velocity and deformation data, including peak systolic and early and late diastolic myocardial velocities, peak systolic strain rate (SR), and peak systolic strain (epsilon), were calculated in the radial direction in the inferolateral LV wall and in the longitudinal direction in the interventricular septum, the LV anterolateral wall, and the right ventricular (RV) free wall. RESULTS: Higher heart rates and increased LV end-systolic dimensions were seen in patients with DMD compared with controls. Significant decreases in radial and longitudinal peak systolic SR, peak systolic epsilon, and peak systolic and early diastolic myocardial velocities were found in the LV inferolateral and anterolateral walls in patients with DMD. No significant differences in longitudinal function could be found in the interventricular septum or in the RV free wall. CONCLUSION: In young patients with DMD who have global normal systolic function, reductions in systolic deformation parameters as well as reduced early diastolic myocardial velocities can be detected in the anterolateral and inferolateral LV walls. The prognostic significance of these findings warrants further longitudinal follow-up. PMID- 18406575 TI - Clinical usefulness of tissue Doppler imaging in patients with mild to moderate aortic stenosis: a substudy of the aortic stenosis progression observation measuring effects of rosuvastatin study. AB - OBJECTIVE: Although impaired diastolic function is common in aortic stenosis (AS), little is known about the clinical usefulness of tissue Doppler imaging (TDI) to detect diastolic dysfunction in patients with mild to moderate AS. The objective was to describe both conventional and TDI measurements of diastolic function in asymptomatic patients enrolled in the Aortic Stenosis Progression Observation Measuring Effects of Rosuvastatin study, a multicenter study to assess the effect of rosuvastatin on the progression of AS. METHODS: Baseline echocardiography measurements, including left ventricular interventricular septal thickness, posterior wall thickness, cavity dimensions, and ejection fraction were obtained. Conventional Doppler indices, including peak early (E) and late (A) transmitral velocities, E/A ratio, and E-wave deceleration time, were measured from spectral Doppler. Tissue Doppler measurements, including early (E') and late (A') velocities of the lateral annulus, were determined, and E/E' was calculated. RESULTS: The study population included 172 patients (aged 57 +/- 13 years; 73 were female) divided into three categories of AS severity on the basis of peak velocity at baseline (group I: 2.5-3.0 m/s; group II: 3.1-3.5 m/s; group III; 3.6-4.0 m/s). Baseline hemodynamics, left ventricular dimensions, and conventional diastolic functional parameters were similar among all 3 groups. In patients with greater severity of AS, the lateral E' was lower and the E/E' (as an estimate of increased left ventricular end-diastolic pressure) was higher (P <.05). CONCLUSION: In patients with mild to moderate asymptomatic AS, TDI measures of diastolic function are abnormal and related to the severity of AS. These findings may help to predict the future development of symptoms in this population. PMID- 18406574 TI - Osteopontin promoter polymorphism is associated with increased carotid intima media thickness. AB - BACKGROUND: Osteopontin (OPN)-transgenic mice exhibit increased carotid artery intima-media thickness (CIMT), smooth muscle cell proliferation, and atheroma formation. METHODS: An association of the human T-66G promoter variant with CIMT was examined in Caucasian adults grouped according to metabolic syndrome criteria: present (+MetS; n = 70) or absent (-MetS; n = 70). RESULTS: The G allele frequency was 22%. For the entire cohort, the G group (TG and GG) was associated with significantly lower age-adjusted and gender-adjusted CIMT compared with the TT group (P = .008); similar analysis by metabolic syndrome group found a significant difference only in the -MetS group (P = .018). Stepwise multivariate regression showed that after age and waist circumference, the T-66G variant was the next most predictive of CIMT (P = .007). These data suggest that in a normoglycemic environment, human vascular OPN gene expression contributes to arterial structure, an effect diminished in dysmetabolic states. CONCLUSION: Humans with the OPN -66 TT genotype, particularly those without metabolic syndrome, exhibit thicker CIMT. PMID- 18406576 TI - CD127 immunophenotyping suggests altered CD4+ T cell regulation in primary progressive multiple sclerosis. AB - Aberrant regulatory T cell populations, characterised by a wide array of CD markers, have been identified in many autoimmune diseases. CD127 has recently been identified as a specific marker for the CD4(+)CD25(Hi) (Tregs) subset. CD127 is the first non-HLA gene to have its association with multiple sclerosis widely replicated. We demonstrate that the regulatory or suppressor T cells CD4(+)CD25(Hi) (Tregs), CD8(+)CD28(-), and CD3(+)CD56(+) (NKT) all produce low levels of CD127, and so could be at a disadvantage in survival and/or proliferation where IL7 is limiting. The remissions seen in relapsing remitting multiple sclerosis (RRMS) could be driven by regulatory T cells, and the absence of remissions seen in primary progressive MS (PPMS) may point to a particularly reduced function of this cell subset. We found that the proportions of CD4(+)FoxP3(+)CD25(Hi) regulatory T cells were not aberrant in PPMS. There was, however, a trend towards reduced FoxP3 expression per cell in this fraction (p<0.083), which has been highly correlated with suppressor function. Notably, we found that the target of regulatory T cells, the CD4(+)CD25(-) cells, was in excess (p<0.009); and in PPMS a protective CD127 haplotype is correlated with higher CD127 expression (p<0.01). These data support further investigations into the regulatory T cell immunophenotype in MS. PMID- 18406577 TI - Differential involvement of Galpha16 in CC chemokine-induced stimulation of phospholipase Cbeta, ERK, and chemotaxis. AB - Chemokines are known to regulate the chemotaxis of leukocytes and play an important role in immunological processes. Chemokine receptors are widely distributed in hematopoietic cells and are often co-localized with the hematopoietic-specific G(16) and its close relative, G(14). Yet, many chemokine receptors utilize pertussis toxin-sensitive G(i) proteins for signaling. Given that both G(16) and G(14) are capable of linking G(i)-coupled receptors to the stimulation of phospholipase Cbeta, we examined the capacity of six CC chemokine receptors (CCR1, CCR2a, CCR2b, CCR3, CCR5 and CCR7) to interact with G(14) and G(16) in a heterologous expression system. Among the CC chemokine receptors tested, CCR1, CCR2b, and CCR3 were capable of mediating chemokine-induced stimulation of phospholipase Cbeta via either G(14) or G(16). The G(14)/G(16) mediated responses exhibited CC chemokine dose-dependency and were resistant to pertussis toxin (PTX) treatment. In contrast, CCR2a, CCR5 and CCR7 were unable to interact with G(14) and G(16). Under identical experimental conditions, all six CC chemokine receptors were fully capable of inhibiting adenylyl cyclase via G(i) as well as stimulating phospholipase Cbeta via 16z44, a G(16/z) chimera that possesses increased promiscuity toward G(i)-coupled receptors. Moreover, CCR1 mediated ERK1/2 phosphorylation was largely PTX-insensitive in THP-1 monocytic cells that endogenously express Galpha(16). In addition, CCR1 agonist was less efficacious in mediating chemotaxis of THP-1 cells following the knockdown of Galpha(16) by overexpressing siRNA, indicating the participation of Galpha(16) in CCR1-induced cell migration. These results show that different CC chemokine receptors can discriminate against G(14) and G(16) for signal transduction. PMID- 18406579 TI - DHMEQ, a novel NF-kappaB inhibitor, suppresses growth and type I collagen accumulation in keloid fibroblasts. AB - BACKGROUND: Keloid is a benign dermal tumor characterized by proliferation of dermal fibroblasts and overproduction of extracellular matrix (ECM). Nuclear factor kappaB (NF-kappaB) plays an important role in regulation of inflammation, immune response and cell proliferation. Activation of the NF-kappaB pathway is thought to be closely linked to abnormal cell proliferation and ECM production in keloid fibroblasts. OBJECTIVE: This study was set out to investigate the effects of a novel selective NF-kappaB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), on keloid fibroblasts. METHODS: Primary normal and keloid dermal fibroblasts were used for this study. NF-kappaB activity was assessed by DNA-binding assay and immunohistochemistry. The effect of DHMEQ was evaluated by cell viability, cell growth and type I collagen accumulation. RESULTS: Basal NF-kappaB activity was constitutively elevated in keloid fibroblasts, indicating that this pathway is involved in keloid pathogenesis. DHMEQ markedly reduced cell proliferation and type I collagen accumulation in keloid fibroblasts. CONCLUSION: The inhibition of NF-kappaB by DHMEQ may be an attractive therapeutic approach for keloids. PMID- 18406580 TI - Anterior cingulate cortex does not differ between patients with major depression and healthy controls, but relatively large anterior cingulate cortex predicts a good clinical course. AB - The anterior cingulate cortex (ACC) is involved in the regulation of emotion processing, and its volume has been found to be reduced in patients with major depression. Furthermore, larger ACC volumes have been associated with faster symptom improvement under therapy. The aims of the study were to examine whether volumes of the anterior cingulate cortex are altered and are related to the clinical course of major depression. Subjects comprised 78 inpatients with major depression and 78 age-, gender- and handedness- matched healthy volunteers, who were investigated with structural magnetic resonance imaging (MRI). The ACC was subdivided into the subgenual, pre-callosal, rostral-anterior and caudal-anterior ACC. No significant differences were observed for ACC volumes between patients and healthy controls. Left ACC volumes showed a significant negative correlation with the number of hospitalizations. These findings suggest that ACC volumes are not altered in patients with major depression, but that patients with larger ACC have a better clinical outcome than patients with smaller ACC. PMID- 18406578 TI - Verbal working memory impairments in individuals with schizophrenia and their first-degree relatives: findings from the Consortium on the Genetics of Schizophrenia. AB - Working memory (WM) impairment is a promising candidate endophenotype for schizophrenia that could facilitate the identification of susceptibility genes for this disorder. The validity of this putative endophenotype was assessed by determining whether 149 probands with schizophrenia and 337 of their first-degree relatives demonstrated WM impairment as compared to 190 unaffected community comparison subjects. Subjects were participants in the Consortium on the Genetics of Schizophrenia (COGS) project, a seven-site research network that was established to investigate the genetic architecture of endophenotypes for schizophrenia. Participants received comprehensive clinical assessments and completed two verbal WM tasks, one requiring transient on-line storage and another requiring maintenance plus complex manipulation of information by reordering the stimuli. Schizophrenia probands performed worse than the other groups on both tasks, with larger deficits found for the more challenging reordering WM task. The probands' relatives performed more poorly than community comparison subjects on both tasks, but the difference was significant only for the more challenging maintenance plus complex manipulation WM task. This WM impairment was not attributable to diagnoses of schizophrenia spectrum disorder, mood disorders, or substance use disorders in the relatives. In conjunction with evidence that WM abilities are substantially heritable, the current results support the validity and usefulness of verbal WM impairments in manipulation of information as endophenotypes for schizophrenia in large-scale genetic linkage and association studies. PMID- 18406581 TI - 1H NMR signal broadening in spectra of alkane molecules adsorbed on MFI-type zeolites. AB - The anisotropic behavior of C1-C6 alkane molecules adsorbed in MFI zeolite was studied by 1H nuclear magnetic resonance (NMR) using single-pulse excitation, Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence, Hahn echo (HE) pulse sequence, and magic-angle spinning. The molecular order parameter was obtained by both static 2H NMR spectroscopy and molecular simulations. This yields an order parameter in the range of 0.28-0.42 for linear alkanes in MFI zeolite, whereas the parameter equals zero for FAU zeolite with a cubic symmetry. Thus, in the case of a zeolite with a non-cubic symmetry like MFI, the mobility of the molecules in one crystallite cannot fully average the dipolar interaction. As a consequence, transverse nuclear magnetization as revealed in the echo attenuation notably deviates from a mono-exponential decay. This information is of particular relevance for the performance of pulsed field gradient (PFG) NMR diffusion experiments, since the occurrence of non-exponential magnetization attenuation could be taken as an indication of the existence of different molecules or of molecules in different states of mobility. PMID- 18406582 TI - [Pathogenesis and laboratory diagnosis of Mycoplasma pneumoniae infections]. AB - Mycoplasma pneumoniae is the only mycoplasma clearly involved in respiratory tract infections in man. Implicated most often in tracheobronchitis, it is the second most frequent agent responsible for community-wide bacterial pneumonia, and in addition it probably causes asthma exacerbations. M. pneumoniae infection occurs endemically, with epidemic peaks every four to seven years, mostly in children above five years of age. The laboratory diagnosis of these infections, mainly by serology, is made only in severe cases because of the fastidious growth of this microorganism. M. pneumoniae can, however, be detected easily by molecular amplification techniques. Macrolides and related antibiotics are considered the treatment of choice for M. pneumoniae infection in both adults and children. Antibiotic sensitivity testing of M. pneumoniae is not done routinely because resistant isolates have only rarely been described, the results are delayed, and they have no immediate therapeutic consequence. PMID- 18406583 TI - Capecitabine, irinotecan, oxaliplatin (CAPIRINOX) and concomitant irradiation in advanced rectal cancer: the Lyon R-02-01 phase I trial. AB - AIMS: To determine the feasibility of radiotherapy-associated capecitabine, irinotecan and oxaliplatin administration at five dose levels for the treatment of locally advanced rectal cancer, with or without metastasis. PATIENTS AND METHODS: This was a bicentric phase I trial, including patients with locally advanced rectal cancer, with or without metastasis. Chemotherapy comprised capecitabine (1100, 1300 or 1500 mg/m2/day, every day), irinotecan (30, 40 or 50mg/m2, once per week for 6 weeks) with the addition of oxaliplatin (40 mg/m2 at level 4 or 50 mg/m2 at level 5, once per week for 6 weeks). Radiotherapy at 46 Gy plus a boost of 4 Gy was administered concomitantly. RESULTS: Twelve patients received four levels of dose. As a supplement to radiotherapy, the combination of capecitabine and irinotecan at the respective doses of 1500 mg/m2/day and 50 mg/m2/week was feasible and well tolerated. The addition of oxaliplatin to this combination provoked toxicity (grade 3/4 vomiting, diarrhoea) for two-thirds of the patients. CONCLUSION: A treatment associating radiotherapy (46 Gy+4 Gy) with concomitant chemotherapy comprising capecitabine (1500 mg/m2/day, every day) and irinotecan (50 mg/m2/week, for 5 weeks) was feasible and well tolerated. The addition of oxaliplatin to these doses was prohibitory to the continuation of treatment due to unacceptable toxicity. PMID- 18406584 TI - A phase II study of oxaliplatin and gemcitabine in advanced inoperable stage IIIB/IV non-small cell lung cancer. PMID- 18406585 TI - Non-invasive diagnosis and monitoring of chronic kidney disease by reverse iontophoresis of urea in vivo. AB - BACKGROUND: Reverse iontophoresis uses a small current to extract molecules and ions through the skin. The aim of the study was to determine whether reverse iontophoresis of urea can be used (i) to diagnose and monitor non-invasively chronic kidney disease (CKD), and (ii) to track urea levels closely during a hemodialysis session. METHODS: A current of 0.8mA was applied for 2h in 10 healthy volunteers, in 9 patients with CKD, and in 10 patients undergoing hemodialysis. Urea fluxes extracted by reverse iontophoresis and urea concentrations in the blood were measured. RESULTS: Extracted urea fluxes discriminated healthy volunteers from patients with CKD within 90 min. A non invasive measure of blood urea concentrations can be achieved after 120 min. A urea reservoir in the skin interferes with the extraction and a pre-hemodialysis "depletion" period is required. Mild and transient sensation and erythema induced by iontophoresis were significantly lower in the CKD group. Gelling the formulation of the iontophoresis reservoir gave similar results to those obtained when using a simple aqueous solution. CONCLUSIONS: Reverse iontophoresis can be used to non-invasively diagnose individuals with CKD and to monitor urea concentrations in blood. PMID- 18406586 TI - Physicochemical stability of phospholipid-dispersed suspensions of crystalline itraconazole. AB - The physicochemical stability of an aqueous, phospholipid-based dispersion of itraconazole microcrystals was studied as a model water-insoluble drug suspension. The particle size, phospholipid concentrations, free fatty acid (FFA) content, pH, and zeta potential of two test suspensions were followed over 63 days at 5 and 40 degrees C storage conditions. Hydrolysis of a control suspension containing Lipoid E80 led to rapid FFA formation, pH drop, and subsequent particle aggregation. In the second suspension, sodium oleate used in conjunction with Lipoid E80 significantly enhanced the suspension physicochemical stability. Oleate anions effectively (1) increased the anionic charge of the phospholipid surface layer, (2) buffered the suspension near pH 7, and (3) reduced the specific production of oleic acid as a phosphatidylcholine (PC) degradant. The observed hydrolysis rate constants k(obs) approximately 2 x 10(-7) (Lipoid only) and k(obs) approximately 5 x 10(-8) (Lipoid and oleate) were consistent with the pH dependent behavior reported for saturated soybean PC solutions. Mechanistically, FFA formed initially in the control suspension partitioned to the aqueous phase with limited influence on the phospholipid microenvironment at the itraconazole particle surface. Phospholipid stabilization of water-insoluble drugs was demonstrated with clear benefits from fatty acid anions as co-additives to influence the surface microenvironment, reduce hydrolysis kinetics, and enhance suspension physicochemical stability. PMID- 18406587 TI - Nebulization of nanoparticulate amorphous or crystalline tacrolimus--single-dose pharmacokinetics study in mice. AB - Developing a pulmonary composition of tacrolimus (TAC) provides direct access to the graft in lung transplant offering the possibility of high drug levels. The objective of this study was to investigate the physicochemical and pharmacokinetic characteristics of the nanostructured aggregates containing amorphous or crystalline nanoparticles of TAC produced by ultra-rapid freezing (URF). TAC and lactose (1:1 ratio; URF-TAC:LAC) and TAC alone (URF-TAC) were investigated for pulmonary delivery and compared to unprocessed TAC. X-ray diffraction (XRD) results indicated that URF-TAC was crystalline, whereas URF TAC:LAC was amorphous. In vitro results revealed the superior physiochemical characteristics of both URF formulations compared to unprocessed TAC. The surface area of URF processed TAC was higher (25-29 m2/g) than that of the unprocessed TAC (0.53 m2/g) and subsequently enhanced dissolution rates. In addition, URF TAC:LAC displayed the ability to supersaturate in the dissolution media to about 11 times the crystalline equilibrium solubility. Similar aerodynamic particle sizes of 2-3 microm, and fine particle fraction between 70% and 75% were found in both formulations. The local and systemic pharmacokinetic studies in mice showed similar AUC(0-24), higher Cmax, and lower Tmax for the URF-TAC:LAC compared to the URF-TAC. Nanostructured aggregates containing amorphous or crystalline nanoparticles of TAC were demonstrated to be effectively delivered via nebulization, with similar in vitro and in vivo performances. PMID- 18406588 TI - Tetracycline-induced reproductive toxicity in male rats: effects of vitamin C and N-acetylcysteine. AB - Tetracycline, a broad-spectrum antibiotic employed clinically in the treatment of bacteria infections, is known to cause a number of biochemical dysfunctions and suspected to induce testicular damage to animals and humans, but there is paucity of data on its effect and mechanism of action on the male reproductive system. The present study therefore evaluates its spermatotoxic and testicular toxicity in male rats and the chemoprotective effects of Vitamin C (Vit C) and N acetylcysteine (NAC). Tetracycline was administered orally at the dose level of 28.6 mg/kg body weight per day in two equal divided doses (12h interval). Vit C and NAC were also administered orally to the rats at doses of 200 and 50 mg/kg body weight per day, respectively, for the 14 days of the experiment. While there was no change in the body weights of rats, tetracycline administration caused significant decrease in the relative weights of testis, epididymis and seminal vesicles (P<0.05). Administration of tetracycline caused a reduction in the epididymal sperm motility, percentage of live spermatozoa, sperm count, and an increase in abnormal sperm morphology, as well as induction of adverse histopathologic changes in the testes. While Vit C and NAC significantly mitigated the toxic effect of tetracycline on sperm parameters, the antioxidants did not improve the adverse histopathologic changes induced by antibiotic. Treatment of rats with tetracycline significantly decreased the activities of superoxide dismutase, catalase (CAT), glucose-6-phosphate dehydrogenase, glutathione-S-transferase (GST) and the levels of GSH and serum testosterone, while the activity of gamma-glutamyltranspeptidase and the formation of malondialdehyde (MDA) increased. Both Vit C and NAC significantly attenuated the toxic effects of tetracycline to the antioxidant and testicular marker enzymes as well as markers of oxidative stress. Collectively, the results suggest that therapeutic dose of tetracycline elicits spermatotoxic and testicular toxicity in male rats through induction of oxidative stress. The chemoprotective effects of Vit C and NAC during tetracycline treatment suggest that these antioxidants may find clinical application in cellular damage involving reactive oxygen species (ROS). PMID- 18406589 TI - Effect of total flavonoid fraction of Astragalus complanatus R. Brown on angiotensin II-induced portal-vein contraction in hypertensive rats. AB - The objective of the present study was to examine further the underlying mechanism of the antihypertensive effect of the total flavonoid (TF), extracted from the seed of Astragalus complanatus R.Brown. Renovascular hypertension rats (RHR) were established by the two-kidney one clip (2K1C) method. The effect of TF on the contraction of portal vein was studied in an isolated preparation. The response of portal vein to angiotensin II (Ang II) was expressed as a percentage of the 100 mmol/l KCl induced maximum contraction. We took the dose-response curve of portal vein to Ang II (from 10(-9) to 10(-6) mmol/l) as the control and then observed the change of curve after TF and Valsartan (Ang II receptor blocker) administration. Ang II induced a concentration-dependent increase of the contraction amplitude (maximal increase, 46.53+/-5.15% of 100 mmol/l KCl induced contraction at Ang II 10(-6) mmol/l in RHR). The Ang II-induced portal vein contraction was prevented by TF with a concentration related manner (maximal inhibition amplitude from 46.53+/-5.15% to 22.525+/-4.67% of 100 mmol/l KCl contraction at 10(-6)mmol/l Ang II and 3.12 x 10(-1) mg/l TF in RHR). The effect of TF on Ang II-induced portal vein contraction was similar to Valsartan. These results showed that the antihypertensive action of TF was attributed to the dilation of vessels and is related to the blockade of the Ang II receptor. PMID- 18406590 TI - Sclerocarya birrea [(A. Rich.) Hochst.] [Anacardiaceae] stem-bark ethanolic extract (SBE) modulates blood glucose, glomerular filtration rate (GFR) and mean arterial blood pressure (MAP) of STZ-induced diabetic rats. AB - Studies in our laboratories suggest that Sclerocarya birrea stem-bark ethanolic extract (SBE) has hypoglycemic properties. Accordingly, we investigated the effects of SBE on major complications of diabetes mellitus; blood glucose, renal function and mean arterial blood pressure (MAP) in non-diabetic and STZ-induced diabetic rats. Oral glucose tolerance test responses to various SBE doses (60, 120 and 240 mg kg(-1)) were studied in fasted rats following glucose load (0.86 g kg(-1), p.o.). Rats treated with deionized water (3 ml kg(-1) p.o.), or standard hypoglycemic drugs (insulin, 100 microg kg(-1), s.c.; metformin, 500 mg kg(-1), p.o. or glibenclamide, 500 microg kg(-1), p.o) acted as untreated and treated positive controls, respectively. Blood was collected in non-diabetic rats after 45 min of SBE, metformin or glibenclamide for plasma insulin determination. Acute SBE effects on renal function and MAP were studied in anesthetized rats challenged with hypotonic saline after 3.5h equilibration for 4h of 1h control, 1.5h treatment and 1.5h recovery periods. SBE was added to the infusate during the treatment period. Chronic effects were monitored for 5 weeks in animals daily treated with SBE (120 mg kg(-1) p.o.) while hepatic glycogen concentration was measured at the end of the experimental period. SBE exhibited dose-dependent reduction in blood glucose concentration. SBE and metformin did not affect plasma insulin secretion in non-diabetic rats, while glibenclamide increased plasma insulin concentration. The hypoglycemic effect of SBE treatment was associated with increased hepatic glycogen synthesis. Acute SBE administration did not significantly alter kidney function, but chronic SBE treatment for decreased plasma urea and creatinine concentrations of STZ-diabetic rats with concomitant increase in GFR by comparison with control rats at the corresponding period (0.7+/-0.2 vs. 1.4+/-0.3 ml min(-1)). SBE treatment reduced blood pressure in all groups of animals. The observations suggest that SBE has reno- and cardio protective effects in diabetes mellitus. The current results indicate the basis for SBE use as complementary remedy in diabetes. PMID- 18406591 TI - Emerging concepts in CD8(+) T regulatory cells. AB - CD8(+) T regulatory cells (T regs) are elicited by unique antigen presenting cells during viral infections, by manipulation of co-stimulatory molecules, or in the development of tumors. CD8(+) T regs display antigen-specificity, which is most exquisitely manifested by the HLA-E-restricted cytolytic CD8(+) T regs in MS. There is evidence that some CD8(+) T regs also express organ specificity. In many cases, IFN-gamma is required for either the induction or expression of CD8(+) T regs. CD8(+) T regs can produce suppression directly by killing immune cells or indirectly by co-opting other cells to elaborate end-stage suppressive molecules such as TGF-beta, IL-10, and indoleamine dioxygenase (IDO). PMID- 18406592 TI - Chorioactidaceae: a new family in the Pezizales (Ascomycota) with four genera. AB - Molecular phylogenetic and comparative morphological studies provide evidence for the recognition of a new family, Chorioactidaceae, in the Pezizales. Four genera are placed in the family: Chorioactis, Desmazierella, Neournula, and Wolfina. Based on parsimony, likelihood, and Bayesian analyses of LSU, SSU, and RPB2 sequence data, Chorioactidaceae represents a sister clade to the Sarcosomataceae, to which some of these taxa were previously referred. Morphologically these genera are similar in pigmentation, excipular construction, and asci, which mostly have terminal opercula and rounded, sometimes forked, bases without croziers. Ascospores have cyanophilic walls or cyanophilic surface ornamentation in the form of ridges or warts. So far as is known the ascospores and the cells of the paraphyses of all species are multinucleate. The six species recognized in these four genera all have limited geographical distributions in the northern hemisphere. PMID- 18406593 TI - Revision and reclassification of three Plasmopara species based on morphological and molecular phylogenetic data. AB - Based on the results of morphological and DNA sequence (partial D1-D3/D7-D8 nuLSU and partial nuSSU-ITS1-5.8S rDNA) data, three species of Plasmopara are revised and reclassified. A species of Plasmopara parasitic on Scorzonera, invalidly published several times, is assigned to a new genus and species under Novotelnova scorzonerae. Plasmopara euphrasiae sp. nov. is segregated from P. densa, and P. centaureae-mollis is revised and relegated to synonymy of Bremia centaureae. All taxa are described and illustrated. PMID- 18406594 TI - Identifying optimal regional solid waste management strategies through an inexact integer programming model containing infinite objectives and constraints. AB - The previous inexact mixed-integer linear programming (IMILP) method can only tackle problems with coefficients of the objective function and constraints being crisp intervals, while the existing inexact mixed-integer semi-infinite programming (IMISIP) method can only deal with single-objective programming problems as it merely allows the number of constraints to be infinite. This study proposes, an inexact mixed-integer bi-infinite programming (IMIBIP) method by incorporating the concept of functional intervals into the programming framework. Different from the existing methods, the IMIBIP can tackle the inexact programming problems that contain both infinite objectives and constraints. The developed method is applied to capacity planning of waste management systems under a variety of uncertainties. Four scenarios are considered for comparing the solutions of IMIBIP with those of IMILP. The results indicate that reasonable solutions can be generated by the IMIBIP method. Compared with IMILP, the system cost from IMIBIP would be relatively high since the fluctuating market factors are considered; however, the IMILP solutions are associated with a raised system reliability level and a reduced constraint violation risk level. PMID- 18406595 TI - Encapsulation of lead from hazardous CRT glass wastes using biopolymer cross linked concrete systems. AB - Discarded computer monitors and television sets are identified as hazardous materials due to the high content of lead in their cathode ray tubes (CRTs). Over 98% of lead is found in CRT glass. More than 75% of obsolete electronics including TV and CRT monitors are in storage because appropriate e-waste management and remediation technologies are insufficient. Already an e-waste tsunami is starting to roll across the US and the whole world. Thus, a new technology was developed as an alternative to current disposal methods; this method uses a concrete composite crosslinked with minute amounts of biopolymers and a crosslinking agent. Commercially available microbial biopolymers of xanthan gum and guar gum were used to encapsulate CRT wastes, reducing Pb leachability as measured by standard USEPA methods. In this investigation, the synergistic effect of the crosslinking reaction was observed through blending two different biopolymers or adding a crosslinking agent in biopolymer solution. This CRT biopolymer-concrete (CBC) composite showed higher compressive strength than the standard concrete and a considerable decrease in lead leachability. PMID- 18406597 TI - A simple and direct electrochemical detection of interferon-gamma using its RNA and DNA aptamers. AB - Tuberculosis is the most frequent cause of infection-related death worldwide. We constructed a simple and direct electrochemical sensor to detect interferon (IFN) gamma, a selective marker for tuberculosis pleurisy, using its RNA and DNA aptamers. IFN-gamma was detected by its 5'-thiol-modified aptamer probe immobilized on the gold electrode. Interaction between IFN-gamma and the aptamer was recorded using electrochemical impedance spectroscopy and quartz crystal microbalance (QCM) with high sensitivity. The RNA-aptamer-based sensor showed a low detection limit of 100 fM, and the DNA-aptamer-based sensor detected IFN gamma to 1 pM in sodium phosphate buffer. With QCM analysis, the aptamer immobilized on the electrode and IFN-gamma bound to the aptamer probe was quantified. This QCM result shows that IFN-gamma exists in multimeric forms to interact with the aptamers, and the RNA aptamer prefers the high multimeric state of IFN-gamma. Such a preference may describe the low detection limit of the RNA aptamer shown by impedance analysis. In addition, IFN-gamma was detected to 10 pM by the DNA aptamer in fetal bovine serum, a mimicked biological system, which has similar components to pleural fluid. PMID- 18406598 TI - Electrochemically fabricated polypyrrole nanofiber-modified electrode as a new electrochemical DNA biosensor. AB - A new biosensor employing immobilized DNA on a nano-structured conductive polymer fixed onto a platinum electrode is presented. Upon optimization of synthesis parameters, polypyrrole nanofibers, 30-90 nm in diameter, were synthesized in an aqueous media by the electropolymerization of pyrrole using normal pulse voltammetry (NPV). The nanofiber film was investigated by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Double-stranded DNA was physisorbed onto the PPy nanofiber films. Various parameters, including the pH and DNA concentration, were optimized. The DNA immobilized on the nanofiber films was characterized using differential pulse voltammetry (DPV) and Fourier-transform infrared (FTIR) spectroscopy. Using DPV to study the interaction of spermidine with DNA, a binding constant (K) value of 4.08 x 10(5)+/-0.05 M(-1) was obtained. For the determination of spermidine, the proposed method exhibited a good dynamic range, correlation coefficient (0.05-1.0 microM and 0.9983, respectively) and a low detection limit (0.02 microM), although Ca(2+) ions were found to electrostatically bind to DNA and weaken the spermidine-DNA interaction. PMID- 18406599 TI - The disulfide bond formation (Dsb) system. AB - In oxidative folding of proteins in the bacterial periplasmic space, disulfide bonds are introduced by the oxidation system and isomerized by the reduction system. These systems utilize the oxidizing and the reducing equivalents of quinone and NADPH, respectively, that are transmitted across the cytoplasmic membrane through integral membrane components DsbB and DsbD. In both pathways, alternating interactions between a Cys-XX-Cys-containing thioredoxin domain and other regulatory domain lead to the maintenance of oxidized and reduced states of the specific terminal enzymes, DsbA that oxidizes target cysteines and DsbC that reduces an incorrect disulfide to allow its isomerization into the physiological one. Molecular details of these remarkable biochemical cascades are being rapidly unraveled by genetic, biochemical, and structural analyses in recent years. PMID- 18406600 TI - Membrane proteins: molecular dynamics simulations. AB - Molecular dynamics simulations of membrane proteins are making rapid progress, because of new high-resolution structures, advances in computer hardware and atomistic simulation algorithms, and the recent introduction of coarse-grained models for membranes and proteins. In addition to several large ion channel simulations, recent studies have explored how individual amino acids interact with the bilayer or snorkel/anchor to the headgroup region, and it has been possible to calculate water/membrane partition free energies. This has resulted in a view of bilayers as being adaptive rather than purely hydrophobic solvents, with important implications, for example, for interaction between lipids and arginines in the charged S4 helix of voltage-gated ion channels. However, several studies indicate that the typical current simulations fall short of exhaustive sampling, and that even simple protein-membrane interactions require at least ca. 1 micros to fully sample their dynamics. One new way this is being addressed is coarse-grained models that enable mesoscopic simulations on multi-micros scale. These have been used to model interactions, self-assembly and membrane perturbations induced by proteins. While they cannot replace all-atom simulations, they are a potentially useful technique for initial insertion, placement, and low-resolution refinement. PMID- 18406601 TI - Theory and simulation. Editorial overview. PMID- 18406602 TI - Sun exposure prior to diagnosis is associated with improved survival in melanoma patients: results from a long-term follow-up study of Italian patients. AB - We followed up 260 melanoma patients included in a population-based case-control study in Turin, Italy. We collected information on host factors and sun exposure history, and analysed their relative survival. Intermittent sun exposure was inversely associated with the risk of death (hazard ratios, HR=0.41 95% confidence interval, CI=0.17-0.98). Outdoor work was not associated with an increased risk of death. Multivariate models including anatomic site, melanoma thickness and histology, showed that intermittent sun exposure had a tendency to be inversely associated with the risk of death from melanoma with a HR of 0.60 (95%CI=0.24-1.5) in the patients with 1 to 59 weeks and a HR of 0.54 (95%CI=0.23 1.2) in patients with more than 60 weeks spent on the beach during their lifetime. This study, with similar methods and a longer follow-up, confirms the finding that sun exposure prior to diagnosis of melanoma is associated with improved survival. PMID- 18406604 TI - Vaccine strategies in the treatment of low-grade non-Hodgkin lymphoma. AB - Recent years have witnessed the development of a variety of promising immunotherapies for treating patients with B-cell non-Hodgkin's lymphomas. Each B lymphocyte expresses an immunoglobulin molecule that is the product of a unique combination of gene segments. B cell malignancy arises from one original B lymphocyte, and therefore all the members of a given lymphoma tumor population have the same unique immunoglobulin, which can serve as a target for immune therapy. When the idiotype (Id), or unique portion, of each immunoglobulin is used as a vaccine, antibodies and T cells can be induced and each can cause rejection of the tumor by the host. This special opportunity for tumor specificity is accompanied by the challenge of constructing a different vaccine for each patient. The first clinical trial of Id vaccination for lymphoma was initiated at Stanford University in 1988. Tumor cells obtained from lymph node sampling were fused with a myeloma cell line to generate a "hybridoma" producing large quantities of idiotype protein. Purified Id protein was then chemically coupled to keyhole limpet hemocyanin (KLH) and emulsified in an "oil-in-water" type immunologic adjuvant. The initial trial included patients with low-grade, follicular lymphoma, in first remission following chemotherapy. Among the first 32 vaccinated patients, roughly half (14/32) developed anti-Id immune responses. These were principally humoral responses rather than cellular responses. Long term follow-up of these 32 patients has revealed that the development of an immune response is strongly correlated with prolonged freedom from disease progression interval and overall survival. Further trials have confirmed significant clinical benefit following Id vaccination. There is reason for excitement about the prospects for effective vaccine therapies for lymphoma as randomized Id vaccine trials commence and newer cell-based vaccine trials enter the clinic. As the clinical activity of lymphoma vaccines becomes established, it will be important to determine how to best integrate active vaccination approaches with standard therapeutic approaches. PMID- 18406603 TI - The ins and outs of GPR30: a transmembrane estrogen receptor. AB - Estrogen is an important hormone in human physiology. It acts both via transcriptional regulation as well as via modulation of intracellular signaling through second messengers. Although estrogen's transcriptional effects occur through classical nuclear steroid receptors (ERs), recent studies reveal the existence of a novel 7-transmembrane G protein-coupled receptor, GPR30, which responds to estrogen and tamoxifen stimulation with rapid cellular signaling including ERK activation, PI3K activation, calcium mobilization and cAMP production. To distinguish between ER- and GPR30-mediated signaling, we have identified a novel GPR30 agonist that exhibits high specificity for GPR30. In this review, we will describe recent work to further our understanding of the role of GPR30 in estrogen biology. PMID- 18406605 TI - Ultrasound-assisted hydrolysis and acidogenesis of solid organic wastes in a rotational drum fermentation system. AB - The hydrolysis and acidogenesis of solid organic wastes in a rotational drum fermentation system (RDFS) were improved by direct ultrasonic irradiation (DUSI) and a modified ultrasonic treatment (MUST) composed of dilution, ultrasonic irradiation, and filtration. The effect of DUSI on VA desorption from particle surfaces was estimated. DUSI delivered few distinctions from the broth characteristics, but elevated pH and VS degradation rate (53% higher than the control) in the subsequent acidogenesis. The results demonstrated that DUSI could dislodge VA from particle surfaces and disrupt large-size particles by hydro mechanical shear force. To improve VA desorption and removal, a MUST process was constructed. The influences of MUST on the characteristics of the fermentation broth and the subsequent acidogenic performance were investigated. MUST raise the broth pH level from 5.1 to 5.5 and remarkably decreased VA concentration from 11.0 to 3.5g/L. At the end of the subsequent acidogenesis, VA increasing ratios, VS degradation ratios, and surface based hydrolysis constants of the fermentors with the control broth (CF) and the treated broth (MUSTF) were 166.7% and 732.0%, 17.0% and 26.7%, and 16.9% and 26.8x10(-6)kgm(-2)d(-1), respectively. With the assistance of MUST, a considerably improved acidogenic performance of solid organic wastes was accomplished in terms of VA production, VS degradation, and particle hydrolysis. PMID- 18406606 TI - Organic matter humification in olive oil mill wastewater by abiotic catalysis with manganese(IV) oxide. AB - The chemical changes occurring in an olive oil mill wastewater (OMW) sample digested catalytically with MnO2 for 30 and 60 days were evaluated comparatively with those occurring in the same OMW left standing for the same time in an open air lagoon. Both treatments increased the pH and electrical conductivity and decreased the contents of dry matter, total organic C and total N, and C/N ratio of OMW. The humic acid (HA)-like fraction isolated from the fresh OMW was characterized by a marked aliphatic character, small O and acidic functional group contents, marked presence of proteinaceous materials, partially modified lignin moieties and polysaccharides-like structures, extended molecular heterogeneity, and small degrees of aromatic ring polycondensation, polymerization and humification. With increasing the time of either lagooning or catalytic digestion, a loss of aliphatic materials and an increase of extraction yield, oxygenation, acidic functional groups, carbohydrates and aromaticity occurred in the HA-like fractions. The more evident changes measured for the HA like fractions from catalytically-digested OMW, with respect to those from lagooned OMW, indicated that MnO2 was able to catalyze organic matter humification in OMW. PMID- 18406607 TI - Characterization of Trametes versicolor laccase for the transformation of aqueous phenol. AB - Laccase (oxygen oxidoreductase, EC 1.10.3.2) from Trametes versicolor was thoroughly characterized in terms of its catalytic stability and its effectiveness as a biocatalyst under various reaction conditions when using phenol as a model substrate. This enzyme demonstrated high or moderate degrees of stability at pHs from 5 to 8 at 25 degrees C and at temperatures from 10 to 30 degrees C at pH 6. Exponential decay expressions were successfully used to model laccase inactivation when incubated under various conditions of pH and temperature. Phenol transformation was optimum at pH 6, but significant transformation was observed over a pH range of 4-7, provided that sufficient laccase was present in the reacting solution. Partial inactivation of laccase was observed during the oxidation of phenol, even under conditions of optimal stability (pH 6 and 25 degrees C). PMID- 18406608 TI - Thermal conversion of elephant grass (Pennisetum purpureum Schum) to bio-gas, bio oil and charcoal. AB - Elephant grass is an abundant, fast growing plant with significant potential as a renewable energy source and for conversion to higher calorific value fuels. This work investigates thermal conversion of elephant grass to bio-gas, bio-oil and charcoal under two heating rates of 10 and 50 degrees C/min. The energy required to pyrolyse elephant grass was evaluated using computer aided thermal analysis technique, while composition of the resultant bio-gas and bio-oil products were monitored with gas chromatographic and mass spectroscopic techniques. At 500 degrees C, the bio-gas compounds consisted primarily of CO2 and CO with small amounts of methane and higher hydrocarbon compounds. The heat of combustion of the bio-gas compounds was estimated to be 3.7-7.4 times higher than the heat required to pyrolyse elephant grass under both heating rates, which confirms that the pyrolysis process can be self-maintained. Faster heating rate was found to increase the amount of liquid products by 10%, while charcoal yields remained almost the same at 30%. The bio-oil mainly consisted of organic acids, phthalate esters, benzene compounds and amides. The amount of organic acids and benzene compounds were significantly reduced at 50 degrees C/min, while the yields of phthalate esters and naphthalene compounds increased. The difference in bio-oil composition with increased heating rate is believed to be associated with the reduction of the secondary reactions of pyrolysis, which are more pronounced under lower heating rate. PMID- 18406609 TI - Use of pulp mill inorganic wastes as alternative liming materials. AB - A laboratory aerobic incubation study was performed during 18 weeks under controlled conditions to assess the effects of applying different doses of pulp mill inorganic wastes on the physical-chemical properties of an acid Dystric Cambissol. Three different inorganic wastes were tested - wood ash, dregs and grits, and an agriculture limestone was used as reference. Results showed that increasing the dose applied of the different inorganic wastes tested always led to significant raises of soil pH at different incubation times demonstrating that its use as alternative-liming materials could be a valid and less expensive option to the use of commercial agricultural limestone. Moreover, no immediate concerns seem to be expected related to soil exchangeable sodium (Na) content, at least for the doses needed to increase soil pH until the targeted value 6.5. Particularly for wood ash a pronounced increase on soil extractable potassium and phosphorous was observed, indicating that besides the liming effect this waste can contribute to improve soil fertility by supplying significant available amounts of these nutrients. Finally, metals do not seem to be a limiting factor for the application to land of these by-products. PMID- 18406610 TI - Isolation of Dunaliella spp. from a hypersaline lake and their ability to accumulate glycerol. AB - The purpose of the present work was to study the potential biotechnological use of Dunaliella species isolated from a hypersaline lake in Turkey. Dunaliella spp. grown in Johnson's medium were isolated and their glycerol production was studied in a batch system in order to determine the optimal conditions required for the highest glycerol accumulation. In the experiments performed with four newly isolated Dunaliella spp., the maximum glycerol accumulation was obtained at 20% NaCl concentration, and pH 6 (for strains T1 and T2) and pH 9 (for strains T3 and T4). Biomass production by strain T2 was significantly higher that by the other strains but the highest glycerol production in broth was obtained by strain T1 followed by strain T2. Strain T1 showed high glycerol production, i.e. 452.57microg/ml of culture broth at 20% NaCl concentration. The highest glycerol accumulation on both dry weight and cell basis was obtained with strain T1, followed by strains T3 and T4 (55.01, 50.16, and 40.23microg/10(6) cells (or pg/cell), respectively) at 25% NaCl concentration. When the high initial inoculum concentration was used at 25% NaCl concentration, strain T1 had the shortest (approximately 10-15days) lag period. This study shows that the isolated strains T1 and T2 can be used for glycerol production because of their high productivity. PMID- 18406611 TI - Olive orchard amended with two experimental olive mill wastes mixtures: effects on soil organic carbon, plant growth and yield. AB - Amendments of olive orchard soil with two different preparations of olive mill solid waste (OMWMs) at the rate of 9tonha(-1) per year for five years in two different plots were compared with an industry standard soil amendment using urea. Both the OMWMs amendments showed significant increases in total organic carbon and humic substances in soil of approximately 40% and 58%, respectively, without negative effects on tree growth and yield. This work has shown that olive oil mill waste (OMW) can be recycled safely using the bioremediation system used in this study. We suggest that this system is particularly beneficial to organic farming and is an alternative solution to direct spreading of raw OMW on farm lands. PMID- 18406612 TI - Optimisation of the anaerobic digestion of agricultural resources. AB - It is in the interest of operators of anaerobic digestion plants to maximise methane production whilst concomitantly reducing the chemical oxygen demand of the digested material. Although the production of biogas through anaerobic digestion is not a new idea, commercial anaerobic digestion processes are often operated at well below their optimal performance due to a variety of factors. This paper reviews current optimisation techniques associated with anaerobic digestion and suggests possible areas where improvements could be made, including the basic design considerations of a single or multi-stage reactor configuration, the type, power and duration of the mixing regime and the retention of active microbial biomass within the reactor. Optimisation of environmental conditions within the digester such as temperature, pH, buffering capacity and fatty acid concentrations is also discussed. The methane-producing potential of various agriculturally sourced feedstocks has been examined, as has the advantages of co digestion to improve carbon-to-nitrogen ratios and the use of pre-treatments and additives to improve hydrolysis rates or supplement essential nutrients which may be limiting. However, perhaps the greatest shortfall in biogas production is the lack of reliable sensory equipment to monitor key parameters and suitable, parallelised control systems to ensure that the process continually operates at optimal performance. Modern techniques such as software sensors and powerful, flexible controllers are capable of solving these problems. A direct comparison can be made here with, for instance, oil refineries where a more mature technology uses continuous in situ monitoring and associated feedback procedures to routinely deliver continuous, optimal performance. PMID- 18406613 TI - N-Acetylhexosaminidase inhibitory properties of C-1 homologated GlcNAc- and GalNAc-thiazolines. AB - Several C-1 homologated GlcNAc- and GalNAc-thiazolines, as well as a related GalNAc-thiazole, have been prepared. The compounds are analogues of GlcNAc thiazoline, a potent transition-state-mimicking inhibitor of retaining beta-N acetylglycosaminidases. Kinetic evaluation of these fused pyranose-heterocycles against the bacterial N-acetylhexosaminidase SpHex suggests active site steric restrictions around the substrate anomeric carbon. PMID- 18406614 TI - Antagonists of the human adenosine A2A receptor. Part 1: Discovery and synthesis of thieno[3,2-d]pyrimidine-4-methanone derivatives. AB - The (-)-(11R,2'S)-enantiomer of the antimalarial drug mefloquine has been found to be a reasonably potent and moderately selective adenosine A(2A) receptor antagonist. Further investigation of this compound has led to the discovery of a series of keto-aryl thieno[3,2-d]pyrimidine derivatives, which are potent and selective antagonists of the adenosine A(2A) receptor. These derivatives show selectivity against the A(1) receptor. Furthermore, some of these compounds have been shown to have in vivo activity in a commonly used model, suggesting the potential for the treatment of Parkinson's disease. PMID- 18406615 TI - Acquired multiple Acyl-CoA dehydrogenase deficiency in 10 horses with atypical myopathy. AB - The aim of the current study was to assess lipid metabolism in horses with atypical myopathy. Urine samples from 10 cases were subjected to analysis of organic acids, glycine conjugates, and acylcarnitines revealing increased mean excretion of lactic acid, ethylmalonic acid, 2-methylsuccinic acid, butyrylglycine, (iso)valerylglycine, hexanoylglycine, free carnitine, C2-, C3-, C4-, C5-, C6-, C8-, C8:1-, C10:1-, and C10:2-carnitine as compared with 15 control horses (12 healthy and three with acute myopathy due to other causes). Analysis of plasma revealed similar results for these predominantly short-chain acylcarnitines. Furthermore, measurement of dehydrogenase activities in lateral vastus muscle from one horse with atypical myopathy indeed showed deficiencies of short-chain acyl-CoA dehydrogenase (0.66 as compared with 2.27 and 2.48 in two controls), medium-chain acyl-CoA dehydrogenase (0.36 as compared with 4.31 and 4.82 in two controls) and isovaleryl-CoA dehydrogenase (0.74 as compared with 1.43 and 1.61 nmol min(-1) mg(-1) in two controls). A deficiency of several mitochondrial dehydrogenases that utilize flavin adenine dinucleotide as cofactor including the acyl-CoA dehydrogenases of fatty acid beta-oxidation, and enzymes that degrade the CoA-esters of glutaric acid, isovaleric acid, 2-methylbutyric acid, isobutyric acid, and sarcosine was suspected in 10 out of 10 cases as the possible etiology for a highly fatal and prevalent toxic equine muscle disease similar to the combined metabolic derangements seen in human multiple acyl-CoA dehydrogenase deficiency also known as glutaric acidemia type II. PMID- 18406616 TI - The impact of p53 and p73 on aneuploidy and cancer. AB - Initiation, progression and evasion are sequential steps in cancer formation, with autonomous cell proliferation as a final outcome. Genetic or epigenetic alterations of key regulatory genes of the cell cycle are frequently associated with these phenomena. Recently, chromosomal instability, a long-supposed driving force of tumorigenesis, was associated with dysregulation of mitotic genes, providing advantages to tumor cells. Numerous molecules thus provide a key link in the chain of relationships between chromosomal instability and cancer. Here, we discuss emerging evidence revealing that two p53 family members, p53 and p73, might be key regulatory genes at the heart of the relationship between chromosomal instability and cancer. We argue that the role of members of the p53 family as tumor suppressor proteins, their impact on the control of cellular ploidy, and their newly emerging connection with mitotic checkpoint regulatory genes support the suggestion that p73 and p53 could be two of the missing links among chromosomal instability, the mitotic checkpoint and cancer. PMID- 18406618 TI - Spinal deposits of a benign choroid plexus papilloma. AB - Choroid plexus papillomas (CPP) are rare, benign tumors of the central nervous system. They usually occur in the lateral ventricles in children and the fourth ventricle in adults. Extensive seeding along the neural axis is usually associated with malignant transformation of the tumor. We report a rare case of a 74-year-old patient with a fourth ventricle CPP associated with diffuse spinal deposits. Radiological and histological findings are presented and the relevant literature is discussed. PMID- 18406619 TI - Influence of platelet time activation on articular cartilage growth in the rabbit knee: preliminary study. AB - PURPOSE: The aim of the present article was to study the influence of platelets and different time activation on cartilage growth in articular defects in the rabbit knee. METHODS: Twelve male New Zealand rabbits (12 weeks) were divided in two groups. Under general anaesthesia, a 4 mm diameter and 2 mm deep defect was performed in medial condyles in both knees. The right knee defect was filled with platelet concentrate 5 min after being activated with ClCa in group A, and 2 min afterwards in group B. Platelets were obtained by centrifuging 10 ml arterial blood from the rabbit prior to the surgical procedure. The left knee defect was not filled. Rabbits were sacrificed 6 weeks after surgery. Macroscopic and microscopic studies were performed. RESULTS: In group A, hyaline cartilage was observed in the right knee defect at the end of the experiment in five rabbits. None of the defects of the left knees showed hyaline cartilage growth. In group B, hyaline cartilage was observed in the right knee defect in only one rabbit. Nevertheless, in group B, all rabbits presented better chondral cellularity and regeneration and lower fibrosis in defects treated with platelets than in non treated ones. CONCLUSIONS: This technique for articular defect reconstruction with platelets is simple and easy, and has shown satisfactory results in our study. Platelets may be useful as an autologous source of multiple growth factors for articular defect reconstruction. Nevertheless, this is a preliminary study and further research is required. PMID- 18406620 TI - Thin film and interface properties during ZnO deposition onto high-barrier hybrid/PET flexible substrates. AB - The combination of transparent conductive oxides with high-barrier films deposited onto flexible polymeric substrates is of considerable importance in order to improve the efficiency, lifetime and stability of flexible electronic devices. In this work, ZnO thin films have been deposited onto high-barrier hybrid/PET flexible substrates by pulsed DC magnetron sputtering, at room temperature and by applying different power values on the target. The employment of in situ and real-time Vis-fUV (1.5-6.5 eV) spectroscopic ellipsometry allowed the investigation of the growth mechanisms of ZnO thin films as well as the modification procedure in the hybrid's surface. Island growth is dominant during the initial stages of deposition concerning low target power regime, whereas layer-by-layer deposition prevails at the high target power regime. The hybrid's modified layer of approximately 10nm was confirmed by the transmission electron microscopy measurements which additionally revealed a columnar structure of the film with a nanocrystalline morphology. The estimated size of the nanocrystals ( approximately 15 nm and above) was compatible with atomic force microscopy (AFM) measurements. Finally, the evolution of the optical parameters (energy gap and absorption peaks) of the ZnO films during the deposition was similar. PMID- 18406621 TI - Destiny of candidates for heart transplantation in the Eurotransplant heart allocation system. AB - OBJECTIVE: We analyzed the prognosis of candidates for heart transplantation (HTx) after being listed with 'urgent status' for donor heart allocation or after ventricular assist device (VAD) implantation without application for urgent status. METHODS: Urgent status as used in this study refers to both the high urgency (HU) status awarded by Eurotransplant until August 31, 2005 and the urgent (U) status that replaced it from then on. Patients who underwent primary VAD implantation between January 2001 and December 2006 and who were listed as transplantable (T) (group VAD-prim, n=159), and patients listed primarily in urgent status before VAD implantation and/or HTx during the same period (group U prim, n=168) were enrolled in the study. Group U-prim consists of subgroups: group U-HTx (n=123), who underwent primarily HTx in urgent status; group U-VAD (n=25), who underwent primarily VAD implantation in urgent status; patients who died in urgent status before HTx or VAD implantation (n=6); and patients in urgent status without HTx or VAD implantation (n=14). The survival rate in each group was studied. RESULTS: Survival rates after VAD implantation in group VAD prim were comparable to those after urgent status listing in group U-prim (67.0% vs 68.5% for 1-year survival, 56.6% vs 65.8% for 2-year survival, respectively). Actuarial survival after listing for urgent status in group U-HTx was significantly better than that in group U-VAD (73.7% vs 46.0% for 1-year survival, p<0.05, log-rank test). Actuarial survival during mechanical circulatory support after the VAD implantation (censored at HTx or weaning from the device) in group VAD-prim was significantly better than that in group U-VAD (80.7% vs 56.2% for 3-month survival, p<0.001, log-rank test). CONCLUSIONS: In order to receive urgent HTx, HTx candidates may choose urgency listing without primary VAD implantation at the risk of failed donor heart allocation in urgent status. However, the prognosis of the patients in the latter situation is poor. PMID- 18406622 TI - Review of squamous premalignant vulvar lesions. AB - Vulvar squamous cell carcinoma (SCC) develops following two different pathways, which have their own premalignant lesions. In the absence of human papilloma virus (HPV), vulvar SCC can develop in a background of lichen sclerosus (LS), differentiated vulvar intraepithelial neoplasia (VIN) or both. The other pathway leading to vulvar SCC is associated with HPV and the HPV-associated premalignancy is usual VIN. In this review we will discuss the history, epidemiology, aetiology, histology, clinical characteristics, treatment options, malignant potential and prevention strategies of the three squamous premalignant vulvar lesions. PMID- 18406623 TI - Identification of a novel regulatory region in the interleukin-6 gene promoter. AB - Interleukin-6 (IL6) is an important pleiotropic cytokine that is regulated at the transcriptional level. To date, most work on its regulation has focused on a 1.2kb region 5' from the start of transcription, similar to published reports on other cytokine genes. This report demonstrates for the first time that a cytokine gene can be regulated by cis-acting regions much further upstream than previously examined. Comparative genomic analysis showed that a 120 kb region contains blocks of sequence conservation between human and rodent genomes, and that a 15 kb region proximal to the start of transcription contains 10 highly homologous sequence blocks of between 100 and 250 bp. By means of a reporter gene assay, a novel transcriptionally active region located between -5307 and -5202 bp upstream from the start of transcription was identified. Electrophoretic mobility shift assays showed nuclear protein(s) binding to this region, thus raising the possibility that the regulatory activity shown by the reporter gene constructs may be mediated by these proteins. These results suggest that the regulation of IL6 expression involves a much larger upstream region than previously examined and the control of IL6 transcription is likely to be regulated by a complex mechanism of modular cis-regulatory elements. PMID- 18406624 TI - Differential expression of osteoblast and osteoclast chemmoatractants in compression and tension sides during orthodontic movement. AB - Orthodontic tooth movement is achieved by the remodeling of alveolar bone in response to mechanical loading, and is supposed to be mediated by several host mediators, such as chemokines. In this study we investigated the pattern of mRNAs expression encoding for osteoblast and osteoclast related chemokines, and further correlated them with the profile of bone remodeling markers in palatal and buccal sides of tooth under orthodontic force, where tensile (T) and compressive (C) forces, respectively, predominate. Real-time PCR was performed with periodontal ligament mRNA from samples of T and C sides of human teeth submitted to rapid maxillary expansion, while periodontal ligament of normal teeth were used as controls. Results showed that both T and C sides exhibited significant higher expression of all targets when compared to controls. Comparing C and T sides, C side exhibited higher expression of MCP-1/CCL2, MIP-1alpha/CCL3 and RANKL, while T side presented higher expression of OCN. The expression of RANTES/CCL5 and SDF 1/CXCL12 was similar in C and T sides. Our data demonstrate a differential expression of chemokines in compressed and stretched PDL during orthodontic tooth movement, suggesting that chemokines pattern may contribute to the differential bone remodeling in response to orthodontic force through the establishment of distinct microenvironments in compression and tension sides. PMID- 18406625 TI - Reduced activation of intracellular signaling pathways in rat prefrontal cortex after chronic phencyclidine administration. AB - Evidence exists that schizophrenia is characterized by deficits in cell-cell communication and information processing. In the present study, we used the phencyclidine (PCP) animal model of schizophrenia to investigate possible defects in intracellular signaling proteins involved in neuroplasticity. Western Blot analysis has been performed to determine total and phospho-protein levels of extracellular signal-regulated kinases 1/2 (ERK1/2), type II calcium/calmodulin dependent protein kinase (alphaCaMKII) and cAMP-response element binding protein (CREB) in prefrontal cortex (PFC) and hippocampus (HIP) of rat chronically treated with PCP, whereas their mRNA levels were determined by real time RT-PCR. We found reduced levels of P-ERK1/2, P-alphaCaMKII and P-CREB in prefrontal cortex of PCP-treated animals when compared to controls, whereas no effects were observed on total protein or mRNA levels. Conversely, no significant changes were detected on protein levels or mRNA expression in hippocampus. Given the role of ERK1/2, alphaCaMKII and CREB in neuroplastic mechanisms and cell communication, our data suggest that their decreased activation following chronic PCP administration can contribute to cortical defects occurring in schizophrenia, and may therefore represent potential targets for pharmacological intervention. PMID- 18406626 TI - Neuro-immuno-endocrine considerations for cancer development and therapeutics. PMID- 18406627 TI - Software tools for molecular microscopy. AB - In our role as the editors of a special edition of the Journal of Structural Biology published in 1996 and devoted to the development of software tools, we offer our view of past developments and future prospects in this area. The astonishing progress in computer hardware over the past decade has fueled a significant increase in computational power available for the solution of macromolecular structures. At the same time the relatively slow growth and development of the accompanying software reflects the difficulties of developing large, complex and very specialized analytical methods. PMID- 18406628 TI - Modeling of region-specific fMRI BOLD neurovascular response functions in rat brain reveals residual differences that correlate with the differences in regional evoked potentials. AB - The response of the rat visual system to flashes of blue light has been studied by blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI). The BOLD temporal response is dependent on the number of flashes presented and demonstrates a refractory period that depends on flash frequency. Activated brain regions included the primary and secondary visual cortex, superior colliculus (SC), dorsal lateral geniculate (DLG), and lateral posterior nucleus (LP), which were found to exhibit differing temporal responses. To explain these differences, the BOLD neurovascular response function was modeled. A second-order differential equation was developed and solved numerically to arrive at region-specific response functions. Included in the model are the light input from the diode (duty cycle), a refractory period, a transient response following onset and cessation of stimulus, and a slow adjustment to changes in the average level of the signal. Constants in the differential equation were evaluated for each region by fitting the model to the experimental BOLD response from a single flash, and the equation was then solved for multiple flashes. The simulation mimics the major features of the data; however, remaining differences in the frequency dependence of the response between the cortical and subcortical regions were unexplained. We hypothesized that these discrepancies were due to regional-specific differences in neuronal response to flash frequency. To test this hypothesis, cortical visual evoked potentials (VEPs) were recorded using the same stimulation protocol as the fMRI. Cortical VEPs were more suppressed than subcortical VEPs as flash frequency increased, supporting our hypothesis. This is the first report that regional differences in neuronal activation to the same stimulus lead to differential BOLD activation. PMID- 18406629 TI - Dynamic Bayesian network modeling of fMRI: a comparison of group-analysis methods. AB - Bayesian network (BN) modeling has recently been introduced as a tool for determining the dependencies between brain regions from functional-magnetic resonance-imaging (fMRI) data. However, studies to date have yet to explore the optimum way for meaningfully combining individually determined BN models to make group inferences. We contrasted the results from three broad approaches: the "virtual-typical- subject" (VTS) approach which pools or averages group data as if they are sampled from a single, hypothetical virtual typical subject; the "individual-structure" (IS) approach that learns a separate BN for each subject, and then finds commonality across the individual structures, and the "common structure" (CS) approach that imposes the same network structure on the BN of every subject, but allows the parameters to differ across subjects. To explore the effects of these three approaches, we applied them to an fMRI study exploring the motor effect of L-dopa medication on ten subjects with Parkinson's disease (PD), as the profound clinical effects of this medication suggest that fMRI activation in PD subjects after medication should start approaching that of age matched controls. We found that none of these approaches is generally superior over the others, according to Bayesian-information-criterion (BIC) scores, and that they led to considerably different group-level results. The IS approach was more sensitive to the normalization effect of the L-dopa medication on brain connectivity. However, for the more homogeneous control population, the VTS approach was superior. Group-analysis approaches should be selected carefully with consideration of both statistical and biomedical evidence. PMID- 18406631 TI - Mitochondrial phylogeny, taxonomy and biogeography of the silvered langur species group (Trachypithecus cristatus). AB - With a distribution ranging from mainland Southeast Asia to the Sunda region, the silvered langur species group is the most widely distributed species complex of the genus Trachypithecus. However, the systematic classification of its members and the phylogenetic relationships among them are less understood, leading to different classification schemes and proposed distribution zones. To address these issues, we sequenced a 573 bp long fragment of the mitochondrial cytochrome b gene from 115 silvered langurs (68 individuals from known origin). According to our data, five monophyletic clades were detected, which refer to the five taxa auratus, cristatus, germaini, margarita and mauritius. The phylogenetic relationships among them are not well resolved, indicating a radiation-like splitting event, which was estimated to have occurred about 0.95-1.25 mya. Within T. cristatus, two major clades were detected, with one comprising specimens from Sumatra, Borneo and the Natuna archipelago, and the other solely individuals from the Malaysian peninsula. According to our findings, we propose to rank all five taxa as distinct species. While T. auratus, T. germaini, T. margarita and T. mauritius seem to be monotypic, T. cristatus should be split into two subspecies, with the Malaysian form being described as new form here. From a phylogeographic perspective, the species group most likely originated on Java. During the early Pleistocene, its range was expanded to the Malaysian peninsula and to the Southeast Asian mainland. Later on, the Malaysian form colonised further regions of the Sunda region, including Sumatra, Borneo and the Natuna archipelago. PMID- 18406630 TI - Large expansion segments in 18S rDNA support a new sponge clade (Class Demospongiae, Order Haplosclerida). AB - Newly emerging molecular phylogenetic hypotheses involving the sponge Order Haplosclerida (Class Demospongiae) are far removed from traditional views on their classification using morphology. In the new grouping of marine haplosclerid taxa by molecular data all members of one highly supported clade were found to have three large indels in the 18S rRNA gene. These indels were not found in this gene in other marine haplosclerids or in any other demosponges analysed. These indels were found in the variable V4 and V7 region of the gene, had high GC contents and formed stable double stranded helices in the 18S rRNA secondary structure. These indels are very important synapomorphies, provide high support for an alternative taxonomic scheme and could help resolve the phylogeny of this order in conjunction with other phylogenetically informative characters. PMID- 18406633 TI - Chondrogenesis, bone morphogenetic protein-4 and mesenchymal stem cells. AB - OBJECTIVE: As adult cartilage has very limited potential to regenerate, cartilage repair is challenging. Available treatments have several disadvantages, including formation of fibrocartilage instead of hyaline-like cartilage, as well as eventual ossification of the newly formed tissue. The focus of this review is the application of bone morphogenetic protein-4 (BMP-4) and mesenchymal stem cells (MSCs) in cartilage repair, a combination that could potentially lead to the formation of permanent hyaline-like cartilage in the defect. METHODS: This review is based on recent literature in the orthopaedic and tissue engineering fields, and is focused on MCSs and bone morphogenetic proteins (BMPs). RESULTS: BMP-4, a stimulator of chondrogenesis, both in vitro and in vivo, is a potential therapeutic agent for cartilage regeneration. BMP-4 delivery can improve the healing process of an articular cartilage defect by stimulating the synthesis of the cartilage matrix constituents: type II collagen and aggrecan. BMP-4 has also been shown to suppress chondrogenic hypertrophy and maintain regenerated cartilage. Use of an appropriate carrier for BMP-4 is crucial for successful reconstruction of cartilage defects. Due to the relatively short half-life in vivo of BMP-4, there is a need to localize and maintain the delivery of BMP-4 to the injury site. Additionally, the delivery of MSCs to the wound site could improve cartilage regeneration; therefore, the carrier should function both as a cell and a protein delivery vehicle. CONCLUSION: The role of BMP-4 in chondrogenesis is significant, and successful methods to deliver BMP-4, with or without MSCs, to the cartilage defect site are a promising therapy to treat cartilage defects. PMID- 18406634 TI - The effects of glycosaminoglycan content on the compressive modulus of cartilage engineered in type II collagen scaffolds. AB - OBJECTIVE: The current study determined the unconfined compressive modulus of tissue-engineered constructs with varying sulfated glycosaminoglycan (GAG) density produced by goat articular chondrocytes in type II collagen scaffolds prepared with a range of cross-link densities and various times in culture. The purpose of this work is to establish a basis for future studies employing constructs of selected maturity (e.g., 25%, 50%, or 75% normal GAG content) for cartilage repair in vivo. METHODS: Porous scaffolds (8 mm diameter by 2 mm thick) were fabricated from porcine type II collagen by freeze-drying, followed by dehydrothermal treatment and carbodiimide cross-linking. In a pilot study, passage 3 adult caprine articular chondrocytes isolated from one goat were grown in scaffolds with six cross-link densities for 2, 3, 4, and 6 weeks (n=3). The goal was to select scaffold cross-link densities and times in culture that would produce constructs with approximately 25%, 50% and 75% the GAG density of native articular cartilage. Based on the results of the pilot study, chondrocytes from three goats were grown in scaffolds with two cross-link densities for three time periods: 3, 5, and 9 weeks (n=6; one of the cross-link groups was run in quadruplicate). The equilibrium modulus from unconfined compression testing of these samples was correlated with GAG content. RESULTS: There was a notable increase in GAG density with decreasing cross-link density. Histological analysis verified a chondrogenic phenotype and revealed various amounts of GAG and type II collagen-containing cartilage. The correlation between modulus and GAG density had a linear coefficient of determination of 0.60. One group with a mean GAG density of 22 microg/mm(3), which was 140% the GAG density of normal caprine articular cartilage, averaged a compressive modulus of 31.5 kPa, which was 10% of caprine articular cartilage tested in this study. CONCLUSIONS: The GAG density and modulus of tissue-engineered constructs can be controlled by the degree of cross-linking of type II collagen scaffolds and time in culture. PMID- 18406635 TI - Letter to Editor re: Varicose Vein Stripping vs Haemodynamic Correction (CHIVA): a Long Term Randomised Trial. by S. Carandina, C. Mari, M. De Palma, M.G. Marcellino, C. Cisno, A. Legnaro, A. Liboni and P. Zamboni, in Eur J Vasc Endovasc Surg 2008;35:230-7. PMID- 18406637 TI - The prevalence, anatomical correlates and treatment of sleep-disordered breathing in obese children and adolescents. AB - The prevalence of childhood obesity is increasing worldwide. One of the obesity related complications that has received increasing attention in recent years is sleep-disordered breathing. Obese children are at a higher risk of developing sleep-disordered breathing, including habitual snoring, obstructive sleep apnea syndrome and desaturations preceded by central apneas. Both adiposity and upper airway factors, such as adenotonsillar hypertrophy, modulate the severity of sleep-disordered breathing in these children. Adenotonsillectomy seems to be effective against obstructive sleep apnea syndrome in obese children. On the other hand, there are limited data on the effects of weight loss and of treatment with continuous positive airway pressure on the severity of sleep apnea in obese children and adolescents. PMID- 18406632 TI - Comparison of dilated fundus examinations with seven-field stereo fundus photographs in the Veterans Affairs Diabetes Trial. AB - OBJECTIVE: The Veterans Affairs Diabetes Trial (VADT) is a 20-medical center, prospective, randomized study of 1792 Type 2 diabetic individuals primarily aimed at determining whether intensive glycemic control prevents macrovascular events. We report a comparison of fundus photographs and ophthalmologic examination at baseline, permitting an evaluation of multiple settings similar to common clinical practice. RESEARCH DESIGN AND METHODS: A 340-patient subset had both local dilated fundus examinations and centrally read seven-field stereo fundus photographs completed within 60 days of each other (median 28 days). Local examiners were unaware of the stereo photographs. RESULTS: Overall, agreement within one step was 76% and exact agreement between ophthalmoscopy and central gradings of fundus photographs on a five-step retinopathy severity scale was 43% (weighted kappa 0.42, CI 0.35-0.48). In about 90% of disagreements the severity level was higher by photographic grading. The sensitivity for ophthalmoscopy compared to grading of fundus photographs for the detection of any retinopathy was 51% and specificity was 91%. For proliferative diabetic retinopathy (PDR), sensitivity was 61% and specificity 98%. Only one eye was high-risk PDR, and it was detected by both methods. For clinically significant macular edema, these measures were 24% and 98%, respectively. The disagreements were of possible clinical importance in three cases (<1%). CONCLUSION: Most disagreements occurred in eyes rated near the milder end of a category and/or resulted from small differences between the ophthalmoscopic and photographic definitions used in classifying severity. There were reasonably few disagreements of possible clinical significance. PMID- 18406636 TI - Mast cell transcriptional networks. AB - Unregulated activation of mast cells can contribute to the pathogenesis of inflammatory and allergic diseases, including asthma, rheumatoid arthritis, inflammatory bowel disease, and multiple sclerosis. Absence of mast cells in animal models can lead to impairment in the innate immune response to parasites and bacterial infections. Aberrant clonal accumulation and proliferation of mast cells can result in a variety of diseases ranging from benign cutaneous mastocytosis to systemic mastocytosis or mast cell leukemia. Understanding mast cell differentiation provides important insights into mechanisms of lineage selection during hematopoiesis and can provide targets for new drug development to treat mast cell disorders. In this review, we discuss controversies related to development, sites of origin, and the transcriptional program of mast cells. PMID- 18406639 TI - Infectious diseases know no borders: a plea for more collaboration between researchers in human and veterinary vaccines. PMID- 18406638 TI - EglD, a putative endoglucanase, with an expansin like domain is localized in the conidial cell wall of Aspergillus nidulans. AB - Although the process of conidial germination in filamentous fungi has been extensively studied, many aspects remain to be elucidated since the asexual spore or conidium is vital in their life cycle. Breakage and reformation of cell wall polymer bonds along with the maintenance of cell wall plasticity during conidia germination depend upon a range of hydrolytic enzymes whose activity is analogous to that of expansins, a highly conserved group of plant cell wall proteins with characteristic wall loosening activity. In the current study, we identified and characterized the eglD gene in Aspergillus nidulans, an expansin-like gene the product of which shows strong similarities with bacterial and fungal endo-beta1,4 glucanases. However, we failed to show such activity in vitro. The eglD gene is constitutively expressed in all developmental stages and compartments of A. nidulans asexual life cycle. However, the EglD protein is exclusively present in conidial cell walls. The role of the EglD protein in morphogenesis, growth and germination rate of conidia was investigated. Our results show that EglD is a conidial cell wall localized expansin-like protein, which could be involved in cell wall remodeling during germination. PMID- 18406640 TI - The stimulatory effect of insulin-like growth factor-1 on the proliferation, differentiation, and mineralisation of osteoblastic cells from Holstein cattle. AB - This study investigated the effect of exogenous insulin-like growth factor (IGF) 1 on the proliferation and differentiation of osteoblastic cells from Chinese Holstein cattle and the resultant bone nodule formation and mineralisation in vitro. The osteoblastic cells were isolated and cultured, then identified using Giemsa and alkaline phosphatase (ALP) staining methods. The effect of different concentrations of IGF-1 on cell growth was assessed by MTT assay. The ALP activity and osteocalcin (OC) concentration in the osteoblastic cells were measured by a colorimetric assay and a radioimmmunoassay, respectively. Calcium nodules were observed using alizarin red S stain, while the content of matrix calcium was determined by atomic absorption spectrophotometry. Cell proliferation in the cultures was stimulated by IGF-1 at concentrations ranging from 1 to 200ng/mL, with the maximum effect observed at 100ng/mL. This effect was observed from day 1 and peaked at day 5, but decreased at day 7. At concentrations of 10ng/mL and 100ng/mL, IGF-1 significantly induced ALP activity, OC level, matrix calcium content, and nodule formation of the osteoblastic cells by 20-180% (P<0.05 or P<0.01), compared to controls. The results suggested that IGF-1 is an anabolic agent for the proliferation, differentiation, mineralisation and calcium content of dairy cow osteoblasts, and could therefore act as a potential treatment for the metabolic bone diseases in these animals. PMID- 18406641 TI - Recent developments in canine locomotor analysis: a review. AB - Subjective evaluation of canine gait has been used for many years. However, our ability to perceive minute details during the gait cycle can be difficult and in some respects impossible even for the most talented gait specialist. The evolution of computer technology in computer assisted gait analysis over the past 20 years has improved the ability to quantitatively define temporospatial gait characteristics. These technological advances and new developments in methodological approaches have assisted researchers and clinicians in gaining a better understanding of canine locomotion. The use of kinematic and kinetic analysis has been validated as a useful tool in veterinary medicine. This paper is an overview of the kinematic and kinetic analytical techniques of the last decade. PMID- 18406642 TI - Interferon-gamma in the serum and effusions of cats with feline coronavirus infection. AB - The aim of this study was to quantify and compare interferon-gamma (IFN-gamma) concentrations in the serum of clinically normal cats infected with feline coronavirus (FCoV) with its concentration in the sera and effusions of cats with feline infectious peritonitis (FIP), a disease associated with infection with a mutated form of FCoV. Clinically normal FCoV-infected cats living in catteries with a high prevalence of FIP had the highest serum IFN-gamma concentrations. The serum concentration of IFN-gamma was not significantly different in cats with FIP compared with clinically normal FCoV-infected animals living in catteries with a low prevalence of the disease. Moreover, the concentration of IFN-gamma was significantly higher in the effusions than in the serum of cats with FIP, probably due to IFN-gamma production within lesions. These findings support the hypothesis that there is a strong, 'systemic' cell mediated immune response in clinically normal, FCoV-infected cats and that a similar process, albeit at a tissue level, is involved in the pathogenesis of FIP. PMID- 18406643 TI - Pressure profile in the caudal extradural space of standing horses before and after extradural drug administration. AB - The objectives of this study were to measure the pressure in the caudal extradural space of standing horses and to evaluate the usefulness of pressure waves to identify correct needle placement. Caudal extradural pressure was measured in 12 healthy horses. The pressure and any extradural pressure waves were recorded for 3min after puncture, for 1min after testing for lack of resistance (LOR), and for 10min after lidocaine injection. Successful extradural drug administration was confirmed in all horses. The median extradural pressure findings after puncture, after LOR, immediately after injection and 10min after needle placement were -1.60kPa (range -2.27 to 1.33kPa), -0.67kPa (-2.27 to 5.73kPa), 5.00kPa (0.93 to 9.87kPa) and 0.13kPa (-0.67 to 4.53kPa), respectively. Extradural pressure waves were not always present. Extradural space pressure was sub-atmospheric in most horses and extradural injection significantly increased this pressure for up to 10min. Extradural pressure waves had limited usefulness in the confirmation of the correct placement of the needle. PMID- 18406644 TI - Pharmacokinetics after intravenous, intramuscular and subcutaneous administration of moxifloxacin in sheep. AB - The disposition kinetics of moxifloxacin, a fluoroquinolone antibiotic, after intravenous (IV), intramuscular (IM) and subcutaneous (SC) administration was determined in sheep at a single dose of 5mg/kg. The concentration-time data were analysed by compartmental (after IV dose) and non-compartmental (after IV, IM and SC administration) pharmacokinetic methods. Plasma concentrations of moxifloxacin were determined by high performance liquid chromatography with fluorescence detection. Steady-state volume of distribution (V(ss)) and clearance (Cl) of moxifloxacin after IV administration were 2.03+/-0.36L/kg and 0.39+/-0.04L/hkg, respectively. Following IM and SC administration, moxifloxacin achieved maximum plasma concentration of 1.66+/-0.62mg/L and 0.90+/-0.19mg/L at 2.25+/-0.88h and 3.25+/-1.17h, respectively. The absolute bioavailabilities after IM and SC routes were 96.12+/-32.70% and 102.20+/-23.76%, respectively. From these data (kinetic parameters and absence of adverse reactions) moxifloxacin may be a potentially useful antibiotic in sheep. PMID- 18406645 TI - Natalizumab treatment in pediatric multiple sclerosis: a case report. AB - Pediatric multiple sclerosis (MS) with manifestations before 16 years of age occurs in 0.4-10.5% of whole MS population. The initial course of the disease is relapsing-remitting with a relapse rate generally higher than that of adults, less than 3% have a primary progressive form. Some recent reports have shown that Interferon beta (IFNbeta) has a strong effect in reducing the relapse rate in children with MS and is well tolerated. We report a 12-year-old girl with MS and a high relapse rate from the onset. Frequent magnetic resonance imaging (MRI) detected persisting inflammatory activity and increase of lesion burden. She continued to present acute relapses and progression of disability in spite of a treatment with IFNbeta-1a at different dosages and the addition of pulse IV steroid treatment. Then, we opted for Natalizumab treatment, recently approved as a monotherapy for patients with MS who experienced inadequate response to other disease modifying therapies and never used till now in pediatric MS. Our patient showed a complete response to Natalizumab with clinical and MRI suppression of disease activity. PMID- 18406646 TI - LAMA2 stop-codon mutation: merosin-deficient congenital muscular dystrophy with occipital polymicrogyria, epilepsy and psychomotor regression. AB - Merosin-deficient congenital muscular dystrophy (MD) type 1A (MDC1A) is one of the most frequent forms of CMD in Western countries. The classical form, characterized by a total lack of laminin alpha2 chain expression, usually shows severe clinical features; cases with complete laminin alpha2 deficiency and mild phenotype have also been reported, although the mechanisms underlying the lack of genotype-phenotype correlation have not been elucidated. Epilepsy and focal cortical dysplasia-in addition to the classical diffuse white matter abnormalities-have been described in some of these patients associated with cognitive deterioration. We report on a patient with total laminin alpha2 deficiency due to a homozygous stop-codon mutation in the LAMA2 gene, with mild evolution. When 6.9 years old, she developed focal occipital seizures and absence like status when awake, with probable relation to an extensive bilateral occipital micropolygyria. Soon afterwards she lost ambulation and developed cognitive deterioration. Our case confirms that the clinical spectrum of MDC1A is more heterogeneous than previously thought. PMID- 18406647 TI - Stroke in a child with neurofibromatosis type 2. AB - Neurofibromatosis types 1 (NF1) and 2 (NF2) are genetically distinct conditions caused by mutations in tumour suppressor genes that share a number of phenotypic features. Childhood stroke and vasculopathy have been associated with NF1, but not with NF2. We describe a case of brainstem stroke in a child with NF2. PMID- 18406648 TI - Effect of deflazacort on cardiac and sternocleidomastoid muscles in Duchenne muscular dystrophy: a magnetic resonance imaging study. AB - OBJECTIVE: To evaluate the involvement of cardiac and sternocleidomastoid muscles by magnetic resonance imaging (MRI) measurement of T2 relaxation time and the left ventricular systolic function in patients with Duchenne muscular dystrophy (DMD) on treatment with deflazacort and compare them with DMD patients without treatment. SUBJECTS: Seventeen patients with DMD (aged 17-22 years) on treatment with deflazacort for at least 7 years and 17 boys with DMD of younger age (12-15 years) without steroid treatment. All patients were free of cardiac or respiratory symptoms and had normal ECG and Holter monitor examination. METHODS: T2 relaxation time of the myocardium (H), left (SCM-L) and right sternocleidomastoid (SCM-R) muscles and left ventricular systolic function were evaluated by magnetic resonance imaging (MRI) in two groups of DMD patients. Myocardial and sternocleidomastoid muscles T2 relaxation time was calculated using 16 TEs (10-85 msec) and TR at least 2000 ms and T2 maps were created. RESULTS: DMD on deflazacort had higher T2 relaxation time values of the heart and of both sternocleidomastoid muscles (T2H median (range): 47 (41-48) vs. 33 (31 37)ms, p<0.001, T2 SCM-L median (range): 35 (30-37) vs. 23 (20-26)ms, p<0.001, T2 SCM-R median (range): 35 (32-37) vs. 23 (20-27)ms, p<0.001) and left ventricular systolic function (LVEDV median (range): 95 (75-120) vs. 90 (80-105)ml, p=0.03, LVESV median (range): 45 (38-55) vs. 47 (41-51)ml, p=0.81(NS), LVEF median (range): 53% (51-57) vs. 48% (42-51), p<0.001) compared to DMD without treatment. CONCLUSIONS: DMD patients on deflazacort are characterized by better preservation of the T2 relaxation time of myocardium and sternocleidomastoid muscles and better LV systolic function. The duration of this beneficial effect needs to be studied prospectively. PMID- 18406649 TI - Direct measurement of dipole-dipole/CSA cross-correlated relaxation by a constant time experiment. AB - Relaxation rates in NMR are usually measured by intensity modulation as a function of a relaxation delay during which the relaxation mechanism of interest is effective. Other mechanisms are often suppressed during the relaxation delay by pulse sequences which eliminate their effects, or cancel their effects when two data sets with appropriate combinations of relaxation rate effects are added. Cross-correlated relaxation (CCR) involving dipole-dipole and CSA interactions differ from auto-correlated relaxation (ACR) in that the signs of contributions can be changed by inverting the state of one spin involved in the dipole-dipole interaction. This property has been exploited previously using CPMG sequences to refocus CCR while ACR evolves. Here we report a new pulse scheme that instead eliminates intensity modulation by ACR and thus allows direct measurement of CCR. The sequence uses a constant time relaxation period for which the contribution of ACR does not change. An inversion pulse is applied at various points in the sequence to effect a decay that depends on CCR only. A 2-D experiment is also described in which chemical shift evolution in the indirect dimension can share the same constant period. This improves sensitivity by avoiding the addition of a separate indirect dimension acquisition time. We illustrate the measurement of residue specific CCR rates on the non-myristoylated yeast ARF1 protein and compare the results to those obtained following the conventional method of measuring the decay rates of the slow and fast-relaxing (15)N doublets. The performances of the two methods are also quantitatively evaluated by simulation. The analysis shows that the shared constant-time CCR (SCT-CCR) method significantly improves sensitivity. PMID- 18406650 TI - Characterization of two missense variants in the hydroxymethylbilane synthase gene in the Israeli population, which differ in their associations with acute intermittent porphyria. AB - Acute intermittent porphyria (AIP) is an autosomal dominant disorder of heme biosynthesis caused by molecular defects in the hydroxymethylbilane synthase (HMBS) gene. In this study, we report two novel missense sequence variations in the HMBS gene, T59I (C176T) and V215M (G643A), in two patients with clinical symptoms compatible with acute attacks of porphyria. However, only the patient who carried V215M presented with full AIP-affirming biochemical evidence. Both variant proteins were expressed in a prokaryotic system and characterized in vitro. Recombinant T59I and V215M had residual activity of 80.6% and 19.4%, respectively, of that of the wild type enzyme. Moreover, changes in K(m), V(max) and thermostability observed in the recombinant V215M suggest a causal relationship between V215M and AIP. The association between the T59I substitution and AIP is less obvious. Based on our investigation, substitution T59I is more likely to be a mutation with a weak effect than a rare form of polymorphism. This study demonstrates that in vitro characterization of missense variations in the HMBS gene can provide valuable information for the interpretation of clinical, biochemical and genetic data, for establishing a diagnosis of AIP. It also highlights the fact that there are still many aspects to be investigated concerning AIP and corroborates the need to report new data that can help to clarify the genotype-phenotype relationship. PMID- 18406651 TI - Lymphomatous pericardial effusion positive for Mycobacterium tuberculosis by PCR analysis. PMID- 18406652 TI - Statin-induced inhibition of HIV-1 release from latently infected U1 cells reveals a critical role for protein prenylation in HIV-1 replication. AB - Latent infection of human immunodeficiency virus type 1 (HIV-1) represents a major hurdle in the treatment of acquired immunodeficiency syndrome (AIDS) patients. Statins were recently reported to suppress acute HIV-1 infection and reduce infectious virion production, but the precise mechanism of inhibition has remained elusive. Here we demonstrate that lypophilic statins suppress HIV-1 virion release from tumor necrosis factor alpha-stimulated latently infected U1 cells through inhibition of protein geranylgeranylation, but not by cholesterol depletion. Indeed, this suppression was reversed by the addition of geranylgeranylpyrophosphate, and a geranylgeranyltransferase-1 inhibitor reduced HIV-1 production. Notably, silencing of the endogenous Rab11a GTPase expression in U1 cells by RNA interference destabilized Gag and reduced virion production both in vitro and in NOD/SCID/gammac null mice. Our findings thus suggest that small GTPase proteins play an important role in HIV-1 replication, and therefore could be attractive molecular targets for anti-HIV-1 therapy. PMID- 18406653 TI - Electron microscopic analysis of dairy microbes inactivated by ultrasound. AB - Ultrasonication is a non-thermal method of food preservation that has the advantage of inactivating microbes in food without causing the common side effects associated with conventional heat treatments, such as nutrient and flavour loss. The aim of this study was to evaluate the use of ultrasound as an alternative to heat pasteurisation and to assess cell damage using transmission electron microscopy (TEM). Three spoilage microbes, previously isolated from pasteurised milk, were used as "test" microbes. Saline solution (SSS) and UHT milk were used as suspension media and were inoculated with exponential growth phase "test" microbes at a microbial concentration of 1 x 10(4) cfu ml(-1). The samples were subjected to power ultrasound (20 kHz, 750 W), at 100%/124 microm wave amplitude for different time intervals. Both Escherichia coli and Saccharomyces cerevisiae were reduced by >99% (for both suspension media) after ultrasonication and Lactobacillus acidophilus was reduced by 72% and 84% in SSS and milk, respectively. Transmission electron microscope micrographs showed that ultrasonication inflicts extensive microbicidal/microbistatic external and internal damage on all three "test" microbes. In E. coli, sonication-induced emulsification caused the formation of unique minute lipopolysaccharide vesicles from the fragmenting cell envelope. PMID- 18406655 TI - Pedagogy, not (only) anatomy of reasoning. PMID- 18406656 TI - Refractive lens exchange for presbyopia. AB - True presbyopia correction still remains one of the main challenges amongst eye professionals. Many corrective techniques exist including bifocal spectacle correction, monovision techniques both with corrective lenses and surgery, multifocal corneal excimer laser and refractive lens exchange with either accommodating or multifocal lenses. Refractive lens exchange was first described more than 10 years ago and has become more accepted in recent years with advancements in lens technology and improvements in surgical techniques. This article reviews developments in this field and current available lens options in the treatment of presbyopia. PMID- 18406657 TI - Evolution and diversity of green plant cell walls. AB - Plant cells are surrounded by a dynamic cell wall that performs many essential biological roles, including regulation of cell expansion, the control of tissue cohesion, ion-exchange and defence against microbes. Recent evidence shows that the suite of polysaccharides and wall proteins from which the plant cell wall is composed shows variation between monophyletic plant taxa. This is likely to have been generated during the evolution of plant groups in response to environmental stress. Understanding the natural variation and diversity that exists between cell walls from different taxa is key to facilitating their future exploitation and manipulation, for example by increasing lignocellulosic content or reducing its recalcitrance for use in biofuel generation. PMID- 18406658 TI - Evaluation of the micronucleus assay in bone marrow and peripheral blood of rats for the determination of cigarette mainstream-smoke activity. AB - The mammalian in vivo micronucleus assay is widely used as part of the genotoxicity testing battery required during the development of new drugs. As such, the in vivo micronucleus assay has been used in a battery of assays for the assessment of cigarette ingredients or design modifications to help ensure that there is no increase in risk or any new risk introduced by these additions or modifications. The present series of studies was conducted to optimize and evaluate this assay for the assessment of the effects of mainstream smoke on the micronucleus frequency in the bone marrow and peripheral blood of rats. In a first experiment, the optimal conditions for performing the micronucleus assay in these tissues were determined. This was done by use of two compounds known for their micronucleus-inducing activity, i.e., the clastogen cyclophosphamide and the aneugen colchicine. In a second experiment, the effects of tube restraint on untreated control rats were investigated. In a third experiment, the optimal conditions were used to assess the clastogenic/aneugenic activity of cigarette smoke in Sprague-Dawley rats. The rat micronucleus assay in both bone marrow and peripheral blood is able to detect clastogenic and aneugenic activity. The flow cytometric determination of micronucleated cells in rat blood is at least as sensitive as determinations in bone marrow. No statistically significant differences were observed in micronucleus frequencies between rats with and without the additional stress of tube restraint; however, the cautious approach would be to use a fresh-air-exposed group (with tube restraint) as the negative control in inhalation experiments. Using the conditions identified as optimal in the above-mentioned experiments, the micronucleus assay was not able to detect effects induced by smoke from conventional cigarettes. Nevertheless, the micronucleus assay will remain a valuable tool as part of a testing battery used to investigate possible adverse effects related to product modifications. PMID- 18406659 TI - Sixty years of follow-up of Hiroshima and Nagasaki survivors: current progress in molecular epidemiology studies. AB - This article provides an overview of the on-going molecular epidemiology studies among atomic-bomb survivors conducted at the Radiation Effects Research Foundation in Japan. The focus is on: (a) inter-individual variations in sensitivity to radiation-induced somatic mutations (glycophorin A (GPA) mutations) and their potential relevance to differences in susceptibility to radiation-related cancers and (b) the role of specific mutations/rearrangements in radiation-induced thyroid and colorectal cancers. The glycophorin A mutant fractions showed large differences between the survivors at each of the estimated bone marrow doses. Of note is the finding at doses>or=1 Gy; that the slope of the mutant fraction was significantly higher in the 'cancer group' than in the 'non cancer group'. This study provided the basis for validating the use of gammaH2AX and reticulocyte micronucleus assays for evaluating radiosensitivity differences and genetic instability, respectively, in our studies in the coming years. Preliminary results from our molecular oncology studies on adult-onset papillary thyroid cancer provide evidence for the induction of RET/PTC rearrangements and BRAF point mutation (both known to be early stage events in adult-onset papillary thyroid cancer) but with a difference: cases associated with the rearrangements were more frequent at high doses, and developed sooner than those with BRAF mutation. In the case of colorectal cancer, the results suggest that radiation exposure might influence microsatellite instability (MSI) status through MSI related epigenetic and genetic alterations-processes that might occur in the early stage of colorectal carcinogenesis. PMID- 18406660 TI - 4-oxo-2-hexenal, a mutagen formed by omega-3 fat peroxidation: occurrence, detection and adduct formation. AB - The purpose of this review is to summarize our recent studies of a novel mutagen, 4-oxo-2-hexenal. To identify the mutagens formed in a model reaction of lipid peroxidation, linolenic acid methyl ester and hemin were reacted with dG. A 4-oxo 2-hexenal-dG adduct (dG*) was identified in the model reaction mixture. The 4-oxo 2-hexenal (4-OHE) showed mutagenic activity in the Salmonella typhimurium strains TA100 and TA104. 4-OHE reacts with DNA to form dG, dC, and 5-methyl-dC(5-Me-dC) adducts (dG*, dC*, 5-Me-dC*) in vitro. After 4-OHE was orally administered to mice, these adducts were detected in esophageal, stomach and intestinal DNA by liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). We also confirmed the formation of 4-OHE during the heat processing of edible vegetable oil, and during cooking. It was present at an especially high concentration in broiled saury. 4-OHE is probably generated by the oxidation of omega-3 fats. These results provide a warning to humans, who may be exposed to this mutagen. Since 4-OHE induces DNA adduct formation in experimental animal organs, further studies on the carcinogenicity of 4-OHE and the detection of 4-OHE-DNA adducts in human tissue will be required. PMID- 18406661 TI - Transgenerational effects from exposure to environmental toxic substances. AB - Exposure of mouse germ cells to radiation and chemicals results in mutation, malformation, cancer and other adverse effects (e.g., functional disorders) in the offspring, though these findings have not been proven in human studies. Environmental toxic substances such as urethane (ethyl carbamate) which had been injected subcutaneously to 50 million people as a co-solvent of analgesics and dioxin (an endocrine disruptor) have been found to be associated with adverse effects in the progeny of mice after parental exposures. There are some reports on congenital malformations in the progeny of fathers who had been exposed to dioxin. However, these substances have not shown mutagenicity in in vitro assay systems such as bacterial systems even with S9, cell transformation assays, etc., in spite of their potent teratogenicity and carcinogenicity in in vivo systems. Urethane was negative in the mouse specific locus test for germ cell mutations, but elicited a significant response at the same loci in the offspring of mice treated during pregnancy. Further, urethane is a mutagen in Drosophila germ cell tests, specifically inducing point mutations. Dioxin (2,3,7,8-tetrachlorodibenzo p-dioxin) does not induce in vivo somatic mutations in mice and rats. It does not induce chromosomal aberrations when the mouse and/or human sperm are treated, but induces mutations at ESTR (expanded simple tandem repeat) loci in mice at low frequencies and also congenital malformations. In this paper, we first present an overview of the results of our studies on transgenerational effects of these toxic substances, compare the results with those obtained after radiation exposure, and then discuss our subsequent studies to reconcile the problems underlying their mutagenicity, teratogenicity and carcinogenicity. PMID- 18406662 TI - A spectroscopic investigation of the weathering of a heritage Sydney sandstone. AB - Infrared spectroscopy has been employed in a study of the degradation of heritage Sydney sandstone used in St. Mary's Cathedral in Sydney, Australia. Spectra were used to characterise the clay components taken from weathered and unweathered sandstone blocks removed from the Cathedral as part of a restoration programme. Two types of kaolin clays - kaolinite and its polymorph, dickite - have been identified. A higher amount of dickite present in the clay of weathered sandstone indicated that a kaolinite-to-dickite transformation occurs upon weathering. X ray photoelectron spectroscopy was also used to confirm the presence of a more thermally stable polymorph of the kaolinite in the sandstone. PMID- 18406663 TI - Time resolved fluorescence spectroscopy of Eu(III) complexation with alpha hydroxy isobutyric acid. AB - Complexation of Eu(III) with alpha-hydroxy isobutyric acid (HIBA), a model compound of humic acid, has been studied by time resolved fluorescence spectroscopy. The ratio of fluorescence intensity of the two peaks at 616 and 592 nm (I(616/592)) was found to increase with increasing ligand to metal ratio. The I(616/592) data was used to deduce the stability constant of Eu-HIBA complexes of the type ML(i) (i=1-3). The formation of multiple ligand complexes was also corroborated by lifetime data which was found to increase with increasing [HIBA]/[Eu] ratio thus indicating replacement of coordinated water molecules by HIBA. PMID- 18406664 TI - Emerging respiratory agents: new viruses for old diseases? AB - The recent advances in molecular technology have enabled the detection of several new viral agents in specimens collected from the human respiratory tract. Human metapneumovirus was first described in 2001, and is a significant respiratory pathogen, particularly of children. Following the identification of severe acute respiratory syndrome (SARS) associated coronavirus, two other newly detected coronaviruses, NL63 and HKU1, have been linked to respiratory disease in humans. However, identifying a new virus as the causative agent of a specific disease is difficult, and ideally would involve satisfying Koch's postulates. The recently described human bocavirus and polyomaviruses KI and WU have been detected in samples collected from humans with acute respiratory infection, but as yet, have not been conclusively proven to be agents of human disease. We review the new viral agents that have been detected in respiratory samples since 2001, and examine their contribution as agents of human disease. PMID- 18406665 TI - The use of E/Em and the time interval difference of isovolumic relaxation (TIVRT IVRTm) in estimating left ventricular filling pressures. AB - BACKGROUND AND AIMS: The ratio of the transmitral and myocardial early diastolic velocities (E/Em) can be used to estimate LV filling pressures (LVFP). Additionally, the time difference between the onset of E and Em also correlates to LVFP. The aim of this study was to evaluate which of these two indices is the best marker of LVFP in a heterogeneous group of patients during a simultaneous invasive procedure. METHODS AND RESULTS: Thirty two patients were studied. Em and the isovolumic relaxation time (IVRTm) at four segments of the LV were measured using pulsed tissue Doppler echocardiography. Pulsed Doppler echocardiography was used to measure E and IVRT. E/Em and IVRT-IVRTm (T IVRT-IVRTm) were then calculated. Highly significant correlations were found between T IVRT-IVRTm and PCWP at the lateral (r= -0.80, p<0.001) and posterior (r= -0.71, p<0.001) segments whereas only a weak relationship was found between PCWP and E/Em (p<0.05). The sensitivity and specificity of using a negative T IVRT-IVRTm for identifying patients with PCWP >12 mm Hg were 89 and 90%, respectively. CONCLUSION: We found a highly significant correlation between T IVRT-IVRTm and PCWP, which was not seen for E/Em. We propose T IVRT-IVRTm as a stronger predictor of LVFP. T IVRT-IVRTm also seems to correlate to LVFP for many different clinical aetiologies of elevated LVFP. PMID- 18406666 TI - Cooling towers and legionellosis: a conundrum with proposed solutions. PMID- 18406667 TI - Embracing technology: patients', family members' and nurse specialists' experience of communicating using e-mail. AB - This paper reports on a study exploring the usefulness of e-mail as a means of communication between nurse specialists and patients with lung cancer and their families. The study involved two lung cancer nurse specialists and 16 patients and family members who used e-mail with them during the 6-month study period. Data were collected from three sources: (1) e-mail contact between the nurse specialists and patients/family members, (2) patient/family member questionnaire and (3) a focus group/reflective session with the nurse specialists. Quantitative data collected from the e-mails and the questionnaires were analysed descriptively and are presented as summary statistics. Text data from the questionnaires and e-mails were analysed using content analysis. Findings suggest that e-mail can be an effective and convenient means of communication between nurse specialists, and patients and family members. Patients and family members reported high levels of satisfaction with this method of communication. It was found to be quick and easy, and patients and family members were satisfied with both the response and the speed of response from the nurse specialists. Nurse specialists were also positive about e-mail use and found that the benefits of using e-mail with patients/family members outweighed any disadvantages. Further investigation is recommended involving other health care professionals and different patient groups to ensure the safe and appropriate use of e-mail within health care. PMID- 18406670 TI - A stable isotope breath test with a standard meal for abnormal gastric emptying of solids in the clinic and in research. AB - BACKGROUND & AIMS: The aim of this study was to validate a [13C]-Spirulina platensis gastric emptying (GE) breath test (GEBT) with a standardized meal. METHODS: Thirty-eight healthy volunteers and 129 patients with clinically suspected delayed GE underwent measurements at 45, 90, 120, 150, 180, and 240 minutes after a 238 kcal meal labeled test with 100 mg [13C]-S platensis and 0.5 mCi 99mTc. We established normal ranges for scintigraphy with this test meal, intraindividual and interindividual coefficients of variation (COVs), and the ability of the [13C] GEBT breath percent dose excreted *1000 values to predict scintigraphic half-life and to categorize GE as delayed, normal, or accelerated. RESULTS: In health, the 10th and 90th percentiles of half-life for scintigraphic GE with this meal were 52 and 86 minutes; intraindividual COVs for scintigraphy and the GEBT were, respectively, 31% and 27% at 45 minutes, 17% and 21% at 90 minutes, 13% and 16% at 120 minutes, 10% and 13% at 150 minutes, and 8% and 12% at 180 minutes. Interindividual COVs at each time for the [13C] GEBT and scintigraphy were typically approximately 1%-4% lower than intraindividual COVs. Individual breath samples at 45, 150, and 180 minutes predicted GE category; at 80% specificity, 45- and 180-minute samples combined were 93% sensitive to identify accelerated GE, and 150- and 180-minute combined were 89% sensitive for delayed GE. CONCLUSIONS: [13C]-S platensis GEBT is as reproducible as scintigraphy; imprecision with both tests reflects physiologic variation. With 4 breath samples, this method with an off-the-shelf meal is valid to assess GE in clinic and in research. PMID- 18406671 TI - Gastric cancer precursors: Pieces of an intriguing story. PMID- 18406672 TI - The management of common gastrointestinal disorders in general practice A survey by the European Society for Primary Care Gastroenterology (ESPCG) in six European countries. AB - BACKGROUND: Gastrointestinal (GI) disorders account for 10% of all consultations in primary care. Little is known about the management of GI disorders by general practitioners (GP) across different European countries. AIM AND METHODS: We undertook a postal survey of randomly selected samples of GPs in six European countries (UK, Holland, Spain, Greece, Poland, Czech Republic) to determine patterns of diagnosis, management and service use in GI disorders. RESULTS: We received 939 responses, response rate 32%. Over 80% of GPs were aware of at least three national guidelines for gastrointestinal disease. The availability of open access endoscopy ranged from 28% (Poland) to over 80% (Holland, Czech and UK). For uninvestigated dyspepsia the preferred first line management was proton pump inhibitor therapy (33-82%), Helicobacter pylori test and treat (19-47%), early endoscopy (5-32%), specialist referral (2-21%). Regarding irritable bowel syndrome, 23% of respondents were familiar with one or more diagnostic criteria, but between 7% (Netherlands) and 32% (Poland) would ask for a specialist opinion before making the diagnosis. CONCLUSION: The wide variation between GPs both between and within countries partly reflects variations in health care systems but also differing levels of knowledge and awareness, factors which are relevant to educational and research policy. PMID- 18406668 TI - Global and local modulatory supply to the mushroom bodies of the moth Spodoptera littoralis. AB - The moth Spodoptera littoralis, is a major pest of agriculture whose olfactory system is tuned to odorants emitted by host plants and conspecifics. As in other insects, the paired mushroom bodies are thought to play pivotal roles in behaviors that are elicited by contextual and multisensory signals, amongst which those of specific odors dominate. Compared with species that have elaborate behavioral repertoires, such as the honey bee Apis mellifera or the cockroach Periplaneta americana, the mushroom bodies of S. littoralis were originally viewed as having a simple cellular organization. This has been since challenged by observations of putative transmitters and neuromodulators. As revealed by immunocytology, the spodopteran mushroom bodies, like those of other taxa, are subdivided longitudinally into discrete neuropil domains. Such divisions are further supported by the present study, which also demonstrates discrete affinities to different mushroom body neuropils by antibodies raised against two putative transmitters, glutamate and gamma-aminobutyric acid, and against three putative neuromodulatory substances: serotonin, A-type allatostatin, and tachykinin-related peptides. The results suggest that in addition to longitudinal divisions of the lobes, circuits in the calyces and lobes are likely to be independently modulated. PMID- 18406673 TI - Mechanical characterization of contact lenses by microindentation: Constant velocity and relaxation testing. AB - Non-destructive methods for testing material properties allow for multiple tests to be performed on the same sample, which will speed up the design and testing process for hydrogel contact lenses. The mechanical properties of contact lenses were investigated by microindentation testing. Indenter force responses were recorded for two modes of testing: constant velocity and relaxation indentation. From these tests, we characterized the biphasic properties of a hydrogel contact lens: Young's modulus of the solid matrix and hydraulic permeability. Measured indenter force response was fit to finite element (FE) simulation results over a range of Young's modulus (E) and hydraulic permeability (k) over a short testing time scale (2s). Estimated hydraulic permeability, 1-5x10(-15)m(4) (Ns)(-1), was similar to previously measured values for Etafilcon A. However, values determined for Young's modulus, 50-60kPa, were lower than previously measured. PMID- 18406674 TI - The comparison of preperitoneal and Lichtenstein repair for incarcerated groin hernias: a randomised controlled trial. AB - BACKGROUND: Lichtenstein hernioplasty can be performed successfully as an emergency operation for incarcerated inguinal hernia. The aim of the study was to compare the short-term and long-term outcomes of the preperitoneal mesh with the Lichtenstein mesh technique in strangula groin hernia. METHODS: Forty consecutive patients with strangulated inguinal hernia were randomized (according to a random table) to undergo either a preperitoneal or a Lichtenstein repair under general anesthesia. Early outcome measures were age, gender, duration of surgery, operating time (min), side of hernia, other pathology, contents of hernia sac, the ratio of the bowel resection, required laparatomy, complete release of the intestinal loop and postoperative complications, time to return to work, driving and full activity. Long-term outcome measures were recurrence. A Student's t-test and Chi-square analysis were used for statistical analysis. RESULTS: They were randomly allocated to undergo either a preperitoneal mesh repair (n=19) or a tension-free mesh repair Lichtenstein (n=21). There were no persistent complications. Mean duration of surgery in the preperitoneal group was 54 min (SD - 11) versus 50 min in the Lichtenstein group (SD - 8). There was no significant difference with regards to age, race, gender, or comorbidities between the 2 groups. Four of the 21 patients (10.5%) who required an additional incision developed some type of complication. This circumstance was found to have significant influence on morbidity (P=0.003) but not on mortality. The median follow-up for the study was 24 months. Patients were seen 1 to 2 weeks after surgery. CONCLUSIONS: In conclusion we recommend preperitoneal repair in strangulated hernia instead of Lichtenstein repair. The use of preperitoneal hernia repair for strangulated inguinal hernia is safe, and any need for laparatomy if bowel resection is necessary. PMID- 18406676 TI - Embryology of the pituitary gland. AB - For some 40 years, the development of the pituitary gland has been the subject of numerous studies. Several aspects of the origin, differentiation, and commitment of the pituitary cells, however, are still in doubt. The pituitary gland of several species apparently originates from the anterior ridge of the neural plate. Glandular pituitary cells can be committed very soon in embryonic life, as early as at the open neural stages. Numbers of differentiation and transcription factors may then control the expression of a given phenotype from the committed cells. Many questions remain unanswered about factors underlying such cellular commitment. These issues call for further studies to elucidate the molecular mechanisms of known and unknown factors controlling the embryology of the pituitary gland. PMID- 18406677 TI - Hanseatic endocrine conference on tissue prorenin-renin-angiotensin systems Local regulatory actions in reproductive and endocrine organs, Hamburg, July 15-16, 1994. AB - Over the past decade, substantial evidence has become available implicating tissue prorenin-renin-angiotensin systems (PRAS) in the local regulation of differentiated cell functions within the tissues where it is expressed. In contrast to a mixed enzyme-endocrine role classically ascribed to the renal renin angiotensin system in the context of cardiovascular homeostasis, the tissue PRAS appears to play an autocrine, paracrine, or even an intracrine regulatory role. It has been well documented that PRAS is expressed in a wide variety of tissues, including reproductive and endocrine glands. We are now poised to take major strides toward understanding such fundamental issues as, for example, the tissue specific regulation of the expression of PRAS, the nature of signal transduction pathways associated with various angiotensin II (AII) receptors, and the relevance of an aberrant expression of the tissue PRAS to pathophysiology encountered in a number of endocrine and reproductive organs. This symposium was organized as a satellite to the Third European Endocrinology Congress, Amsterdam, The Netherlands, to focus the discussion on current advances in these important aspects of tissue PRAS and to generate ideas on future research strategies. PMID- 18406675 TI - The pathogenesis of the bone disease of multiple myeloma. AB - Multiple myeloma is a fatal hematologic malignancy associated with clonal expansion of malignant plasma cells within the bone marrow and the development of a destructive osteolytic bone disease. The principal cellular mechanisms involved in the development of myeloma bone disease are an increase in osteoclastic bone resorption, and a reduction in bone formation. Myeloma cells are found in close association with sites of active bone resorption, and the interactions between myeloma cells and other cells within the specialized bone marrow microenvironment are essential, both for tumor growth and the development of myeloma bone disease. This review discusses the many different factors which have been implicated in myeloma bone disease, including the evidence for their role in myeloma and subsequent therapeutic implications. PMID- 18406678 TI - Estrogenic soybean isoflavones and chronic disease Risks and benefits. AB - Many edible plants contain natural estrogens called phytoestrogens. These compounds possess mixed estrogen agonist-antagonist properties that are organ specific in vivo. We have focused on estrogenic soybean isoflavones because of their potential extensive dietary availability. In this article, we review the clinical and experimental evidence for the possible benefits and risks of ingestion of estrogenic isoflavones throughout the life span, and highlight areas needing further elucidation. PMID- 18406679 TI - Alterations in hepatocyte uptake and plasma binding of thyroxine in nonthyroidal illness and caloric deprivation. AB - Low plasma T(3) in severe illness is widely thought to be due principally to inhibition of 5'-deiodinase activity, but other factors also contribute to this response. Abnormal plasma constituents, namely, 3-carboxy-4-methyl-5-propyl-2 furan propanoic acid (CMPF) and indoxyl sulfate in uremia, and elevated bilirubin and nonesterified fatty acids (NEFA) can impair T(4) transport into hepatocytes, thereby contributing to the lowering of plasma T(3). Assessment of possible endogenous or exogenous inhibitors of T(4) binding to plasma proteins is prone to dilution-dependent artifacts, which can lead to overestimation or underestimation of competitor potency, depending on experimental details. Because the potency of such competitors is a function of their free concentrations in undiluted serum, inhibitory activity may be enhanced by substances that impair their albumin binding. Oleic acid or CMPF can inhibit the effect of drugs such as furosemide or fenclofenac. PMID- 18406680 TI - Sexual differentiation of brain and behavior in birds. AB - It is currently accepted that most sex differences in brain and behavior do not result from direct genomic actions, but develop following early exposure to a sexually differentiated endocrine milieu. In Japanese quail (Coturnix japonica), in contrast to rodents, the male reproductive phenotype appears to develop in the absence of endocrine influence, and estradiol secreted by the ovary of the female embryo is responsible for the physiologic demasculinization of females. In zebra finches (Taeniopygia guttata), estrogens administered early in life demasculinize copulatory behavior in males, but masculinize the vocal control regions in the brain and singing behavior of females. It is difficult to understand how these behaviors differentiate given that normal untreated males sing and copulate in a male-typical manner, whereas females never show these behaviors. All attempts to resolve this paradox with experiments based on the rodent model of sexual differentiation have been unsuccessful. We propose that copulatory behavior in zebra finches is differentiated in a manner similar to what has been described in quail, but that novel approaches need to be considered to understand the differentiation of the telencephalic song control system. In particular, the possible involvement of afferent input that may differentiate in a steroid dependent or -independent manner should be thoroughly tested. PMID- 18406681 TI - Insulinlike growth factor binding proteins and tumor hypoglycemia. AB - Hypoglycemia associated with nonislet cell tumors is a relatively rare metabolic disorder, which develops in the presence of low or unmeasurable serum insulin levels. Understanding of the pathogenetic mechanisms has been obscured by the lack of appropriate analytic methods. It now appears that this disorder can be classified as a paraneoplastic syndrome caused by the oversecretion of unprocessed (big) insulinlike growth factor (IGF) II propeptide by the tumor. In contrast to other paraneoplastic syndromes, however, this oversecretion does not lead to grossly elevated IGF II RIA values in the sera of the patients, but is masked by (a) a negative feedback that suppresses the production of mature (7.5 kD) IGF II, so that the total IGF II level remains more or less unchanged and (b) an altered distribution of the hormone between two specific IGF binding protein complexes in the circulation so that its bioavailability is essentially enhanced. PMID- 18406682 TI - Growth hormone-releasing peptide and its analogues Novel stimuli to growth hormone release. AB - GHRPs are oligopeptides with GH-releasing effects in humans when given by either parenteral or oral routes; in addition, nonpeptide pharmacologic analogues have recently been synthesized. Although the exact mechanism of action of these agents has not been fully established, there is probably a dual site of action on both the pituitary and the hypothalamus, possibly involving regulatory factors in addition to GHRH and somatostatin. GHRPs and their analogues may have a potential role in the treatment of short stature in children or in other situations of GH deficiency, such as adult GH deficiency, obesity, catabolic states, and even normal old age. PMID- 18406683 TI - Postmenopausal estrogen therapy, 1995. AB - Estrogen therapy offers women important benefits, including the potential prolongation of life by prevention of coronary heart disease (CHD) and osteoporotic fractures, and improvement of life by prevention of menopausal symptoms. Not all women can accept the potential side effects and risks, however, which include uterine bleeding, cystic mastitis, fluid retention, and increased risk of uterine cancer and breast cancer. Thus, the benefits and risks must be patiently weighed for each individual, and the alternatives of no treatment or other treatment should be clear. We review here the pertinent information that now makes these decisions quite sound. PMID- 18406684 TI - Influence of metabolic substrates and obesity on growth hormone secretion. AB - In addition to stimulating body growth, GH plays an important role in metabolism. In turn, various products of intermediary metabolism, such as glucose, free fatty acids, dietary proteins, and amino acids feed back on both the hypothalamus and the anterior pituitary to control the function of the somatotroph cell. Alterations in nutritional status, such as malnutrition and obesity, markedly influence GH secretion and/or GH actions at tissue level. Therefore, the interaction between metabolic substrates and GH secretion can be viewed as part of the overall regulation of feeding and fasting in order to maintain an adequate body weight and body composition. PMID- 18406685 TI - Growth factors in reproduction. AB - The changes that occur in the female reproductive tract during the menstrual cycle and in early pregnancy, in preparation for embryo implantation and subsequent placental and fetal development, have long attracted the interest of reproductive biologists. The early embryo expresses growth factors and growth factor receptors that are, in general, temporally expressed. In addition, the oviduct and uterine endometrium also synthesize growth factors and cytokines, and the developing embryo finds itself in a growth-factor-rich milieu during early development. The autocrine, juxtracrine, and paracrine actions of these factors in the development of the early embryo and of the maternal reproductive tract are just beginning to be appreciated, as are their potential roles in cellular proliferation and cell-cell communication within the developing embryo, in the reproductive tract, and at the maternal-trophoblast interface. When detailed mechanisms underlying these interactions are fully understood, it is anticipated that therapies will be rationally designed to treat reproductive disorders associated with abnormal embryonic development and poor placentation. PMID- 18406686 TI - Clinical relevance of estrogen receptor variants in breast cancer. AB - It is well accepted that the presence of estrogen receptor (ER) in breast cancer patients correlates with a better prognosis and a higher probability of response to hormonal treatment. Recent data suggest the ER variant isoforms may be common in clinical breast cancer. Furthermore, an association between the expression of such variants and the development of antihormone resistance is discussed. Although several functionally different ER variants have been described, their significance in the prognosis and treatment of breast cancer is still hypothetical. PMID- 18406687 TI - Receptors for sex steroids in the primate corpus luteum New insight into gonadotropin and steroid action. AB - The evidence is compelling for progesterone receptor (PR) expression in the primate corpus luteum during the menstrual cycle, based on three experimental approaches: (a) immunocytochemistry, (b) radioligand binding to steroid-depleted tissue, and (c) reverse transcription-polymerase chain reaction or ribonuclease protection assay. This information is providing the impetus for studies on possible receptormediated roles for progesterone to control periovulatory events (including follicle rupture and luteal development) and the functional lifespan of the corpus luteum. Similar experiments suggest that estrogen receptors are nondetectable in the corpus luteum. Thus, "classic" receptor-mediated actions of estrogen, such as promoting PR expression, are not apparent; rather, the midcycle surge of LH assumes the role of stimulating PR expression in luteinizing granulosa cells. The recent discovery of androgen receptor expression in primate luteal tissue should lead to studies on the heretofore unsuspected actions of androgens in the corpus luteum. PMID- 18406688 TI - Glucocorticoid resistance in humans. AB - Generalized inherited glucocorticoid resistance is a rare disorder caused by glucocorticoid receptor mutations and characterized clinically by hypercortisolism. Pituitary and peripheral resistance are balanced so that neither adrenal insufficiency nor Cushingoid features develop. Clinical characteristics reflect overproduction of adrenal androgens and mineralocorticoids, which are caused by excess stimulation of the adrenal gland by ACTH. These clinical characteristics respond to dexamethasone therapy. Localized and/or acquired glucocorticoid resistance may occur in some circumstances. PMID- 18406689 TI - Hormonal control of spermatogenesis. AB - FSH and testosterone (T) secretion are essential for the successful completion of spermatogenesis. Because there are no receptors for FSH or testosterone on germ cells, there are intermediate steps in this action, the nature of which are unknown. However, as the Sertoli cell contains receptors for both FSH and T, it is likely that these hormones exert their influence on germ cells by modulating Sertoli cell function. Both FSH and T exert synergistic actions on germ cells, but T has a specific action on the later stages of spermatid maturation. FSH, by its ability to stimulate Sertoli cell mitosis during testicular development, can influence the spermatogenic capacity of the adult testis. PMID- 18406690 TI - Inactivation of thyrotropin-releasing hormone (TRH) by the hormonally regulated TRH-degrading ectoenzyme A potential regulator of TRH signals? AB - The peptidergic signal substance TRH is inactivated by the TRH-degrading enzyme, a peptidase that exhibits a high degree of substrate specificity and other unusual characteristics. The tissue-specific regulation of the adenohypophyseal enzyme by estradiol and thyroid hormones suggests that it may serve an integrative function in modulating the response of adenohypophyseal target cells to TRH and thus pituitary hormone secretion. The high enzymatic activity of neuronal cells indicates that centrally this peptidase also might act as a terminator of neural TRH signals. PMID- 18406691 TI - Pulsatile hormone secretion: Analysis and biological significance. AB - Pulsatile hormone release is a general phenomenon that can be observed in numerous endocrine systems. The analysis and biological significance of pulsatile hormone release were discussed at a Ferring satellite symposium of the Third European Congress of Endocrinology, held on July 23 and 24, 1994, in Hannover, Germany. PMID- 18406692 TI - Melanocortins and their implication in melanoma. AB - The melanocortins (MCs), that is, the melanocyte-stimulating hormones (MSHs) and ACTH, are a group of related peptides containing the typical melanotropin core sequence, His-Phe-Arg-Trp, and are derived from a common precursor, pro opiomelanocortin. They are pleiotropic molecules that occur in the pituitary, some brain regions, and also in several peripheral tissues, and they exert a variety of physiologic functions. Their effect on melanogenesis in the skin is well established, but their role in melanocyte and melanoma cell proliferation and metastasis is less clear. The recent cloning of five types of MC receptors (MC1-5), new studies on the regulation of these receptors, the discovery of a naturally occurring MSH antagonist, the agouti protein, and the finding that melanocytes and melanoma cells exclusively express MC1 receptors have laid the basis for the future development of specific MC ligands, which may become useful for melanoma diagnosis and eventually therapy. PMID- 18406693 TI - Functional analysis of mammalian members of the transforming growth factor-beta superfamily. AB - Analysis of growth factor function has come from studies both in vitro and in vivo. However, the generation of mice deficient in a specific growth factor via gene targeting (for example, "knockout") strategies in embryonic stem (ES) cells will often evaluate the essential roles of the protein in vivo and, in many cases, discover new functions. In this review, studies to date are discussed on the generation and analysis of mice deficient in members of the transforming growth factor (TGF-beta) superfamily. Among the genes targeted via ES cell strategies are the TGF-beta1, Mullerian-inhibiting substance (MIS), inhibin alpha, activin betaA, and activin betaB genes. In addition, the mouse short ear and brachypodism mutants and insertional mutant 413-d have been identified as mutations in the BMP-5, GDF-5, and nodal loci, respectively. These studies have led to critical insights into the functions of these gene products and have further emphasized the importance of members of the TGF-beta superfamily in mammalian development, reproduction, and oncogenesis. PMID- 18406694 TI - Pathogenesis and medical management of benign prostatic hyperplasia. AB - Benign prostatic hyperplasia is an almost universal disorder of aging males. The disease is heterogeneous with respect to the histology, size, symptoms, and response to medical management. Current medical therapies are based upon the knowledge of the pathophysiology of the disease and include hormonal therapies and alpha(1)-adrenergic blockade. Although there are at present no prognostic indicators of response to therapy, future studies may help delineate those patients who will most benefit from the various forms of treatment. PMID- 18406695 TI - Unraveling the human progesterone receptor signal transduction pathway Insights into antiprogestin action. AB - The steroid hormone progesterone is a key modulator of the cellular processes that are required for the development and maintenance of reproductive function. Produced primarily by ovarian granulosa cells, it mediates its biological activity throughout the body by interacting with specific high-affinity nuclear receptors located in target cell nuclei. These receptors are latent transcription factors, which, upon binding progesterone, are capable of interacting with specific recognition sequences within target gene promoters. The consequence of these interactions are determined by the cell and promoter context of the DNA bound receptor. Abnormalities in the progesterone receptor (PR) signal transduction pathway are implicated in pathological states such as breast cancer, endometriosis, and uterine fibroids. Consequently, as a result of the medical need to modulate PR transcriptional activity, antiprogestins, compounds that oppose the actions of progesterone, have been developed. Recent advances in our understanding of the molecular mechanism of action of progesterone have revealed the likely mechanisms by which antiprogestins manifest their biological activity. It is anticipated that this new information will facilitate the discovery and development of additional antiprogestins that may demonstrate superior therapeutic profiles. PMID- 18406696 TI - Altered pulsatile gonadotropin signaling in nutritional deficiency in the male. AB - Reproduction cannot occur without adequate nutrition. Diets that are nutritionally inadequate delay and disrupt the pubertal development of the reproductive processes of immature experimental animals and humans, and impair the function of the hypothalamic-pituitary-gonadal axis in adults. Although there is a general understanding of the linkages between nutrition and reproduction, there is a lack of detailed knowledge of the exact mechanisms that couple these two systems. The major effects of malnutrition on the hypothalamic-pituitary gonadal axis reported in the literature are, for the most part, manifested as reduced gonadotropin secretion. Malnutrition results in decreased circulating gonadotropin concentrations. These changes in the reproductive system are associated with impaired gonadal function and subsequent secondary sex organ atrophy and lead, ultimately, to poor reproduction. Decreased hypothalamic release of gonadotropin-releasing hormone (GnRH) has been proposed as the most important etiologic factor for the fasting-induced suppression of pituitary testicular function. In the human, hypogonadism and infertility develop in both sexes during chronic malnutrition. Most studies on the effects of malnutrition on the reproductive hormones have been performed in women, perhaps because malnutrition in women is promptly accompanied by amenorrhea, whereas in men hypogonadism develops gradually and becomes clinically evident only during more severe malnutrition. With the advent of sensitive assays for measuring reproductive hormones and of modern computerized methods for analyzing the pulsatile secretion of these hormones, however, the function of the hypothalamic pituitary-testicular axis has been scrutinized and it has, indeed, been observed that this system is disturbed even during acute malnutrition. Here, we review the effects of malnutrition on reproductive function, especially on the pulsatile pattern of LH secretion, in humans and in experimental animals. PMID- 18406697 TI - The genetic basis of glucocorticoid resistance. AB - Familial glucocorticoid resistance is a rare syndrome characterized by elevated levels of plasma cortisol but lacking the symptoms of Cushing's syndrome. Biochemically, the condition is characterized by a relative resistance to glucocorticoids that can be compensated for by the elevated levels of cortisol. Analysis of mutations within the receptor resulting in relative glucocorticoid resistance, both familial glucocorticoid resistance and directed mutagenesis, has identified two regions of clustered mutations in the proximity of previously identified affinity-labeled residues. In the majority of cases, the mutation affects steroid binding and transactivation to the same degree, but this is not always the case. PMID- 18406698 TI - Molecular control of meiosis. AB - The animal cell cycle consists of a round of chromosomal DNA replication in S phase, followed by segregation of the replicated chromosomes into the daughter nuclei during M-phase. In most animal cells, gap phases termed G(1) and G(2) are introduced between the M- and S-phases, respectively. Meiosis is a particular example of cell division occurring in germ cells. This specialized cell cycle consists of two successive rounds of chromosome segregation that follow a round of DNA replication. Meiosis produces progeny cells with half as many chromosomes as their parents, thus making sexual reproduction possible. This review is concerned with the factors that have been implicated in the control of meiosis, although research in progress may reveal additional regulatory processes. PMID- 18406699 TI - Androgen receptor dysfunction in human androgen insensitivity. AB - The androgen insensitivity syndromes comprise a spectrum of phenotypic abnormalities in male sex differentiation and development that result from target tissue resistance to androgen action due to molecular lesions in the X-chromosome linked androgen receptor gene. The androgen receptor, like other members of the superfamily of steroid receptors, is characterized by the presence of three structural domains that function in transcriptional activation, DNA binding, and steroid binding, respectively. Missense mutations in the androgen receptor gene causing amino acid substitutions are the most common molecular lesions among affected subjects; deletions, mRNA splice site alterations, and nonsense mutations occur less frequently. Because of the large number and diverse array of these naturally occurring mutations and their associated clinical phenotypes, there is a great opportunity for understanding the structure-function relationships of the androgen receptor from in vitro and in vivo expression of the mutant receptors in various cell types. PMID- 18406700 TI - Actions of epidermal growth factor and its receptor in the thyroid. AB - Since its discovery by Stanley Cohen (1962), epidermal growth factor (EGF) has been found to influence the growth and function of most mammalian cells. EGF is secreted, after cleavage of a large precursor molecule, as a 53-amino acid polypeptide that exerts its effects through the epidermal growth factor receptor (EGF-R), a single 170-kD transmembrane molecule exhibiting intrinsic tyrosine kinase activity of crucial importance to signal transduction (Hsuan et al 1989). Although generally mitogenic, EGF has a wide range of other effects, which vary considerably among organs, cell types, and species. [For a comprehensive update, see the review by Fisher and Lakshmanan (1990).] This article summarizes the present knowledge of EGF actions on thyroid follicular cells (thyrocytes), discusses the possible role of EGF in physiological and pathological conditions of the thyroid gland, and points out some issues that warrant further studies. PMID- 18406701 TI - Partially autonomous cortisol secretion by incidentally discovered adrenal adenomas. AB - Recent studies of the function of adrenal "incidentalomas" have revealed that a proportion of those tumors secrete cortisol insufficiently to produce overt clinical Cushing s syndrome, but that their autonomous cortisol production can suppress the hypothalamo-pituitaryadrenal (HPA) axis to various degrees; this needs to be recognized to avoid acute adrenal insufficiency after adrenalectomy. Several diagnostic approaches have been utilized to identify the partially autonomous cortisol-secreting adenomas. It has been suggested that a lack of normal suppression of cortisol (> 140 nmol/L) on the morning after 1-mg oral dexamethasone at bedtime would identify most functional autonomous cortisol secreting tumors. Based on this criterion, approximately 18% of published cases of incidentalomas would secrete cortisol autonomously. However, other tests indicating alterations of the HPA axis, such as abnormal adrenal iodocholesterol uptake or decreased plasma levels of dehydroepiandrosterone sulfate (DHAS), were found to be present in up to 79%-86% of incidentalomas. This is illustrated by the description of three patients with incidentalomas with plasma cortisol levels < 140 nmol/L in 2 of 3 patients after 1-mg dexamethasone overnight; however, various degrees of HPA axis suppression were demonstrated by an i.v. dexamethasone (4-mg) suppression test, decreased plasma DHAS levels and unilateral adrenal iodocholesterol uptake. After laparoscopic adrenalectomy, the response of plasma cortisol to 250 mug i.v. of ACTH (1-24) was subnormal in 2 of 3 patients and was restored to normal within 2 months. We conclude that the criterion of a plasma cortisol level > 140 nmol/L, after an overnight 1-mg dexamethasone suppression test, underestimates the incidence of partially autonomous cortisol-secreting adrenal adenomas. The literature on this subject is reviewed, and recommendations for evaluation and treatment are presented. PMID- 18406702 TI - Nongenomic actions of progesterone on human spermatozoa. AB - Progesterone's (P) stimulatory actions on human spermatozoa have been known for many years. P indeed appears to be the main sperm stimulator present in women's biological fluids, particularly the follicular fluid. The nongenomic nature of the biological effects of P on human spermatozoa has been demonstrated only recently. P and 17-alpha-hydroxy P have been shown to increase sperm intracellular calcium, phosphatidylinositide hydrolysis, and tyrosine phosphorylation of proteins, and to induce the acrosome reaction (AR), through a rapid, nongenomic mechanism. The effect on calcium is due to influx of the ion from the extracellular medium, as it is inhibited by the calcium chelator EGTA and appears to be mediated by P-binding sites present on the sperm surface, particularly at the head level. The nature of such binding sites has not been addressed so far, but the lack of inhibition of P action by the potent antiprogestin RU486 strongly suggests a biochemical difference from the genomic ones. Evidence exists for involvement of the phospholipid platelet-activating factor (PAF), polyamines, tyrosine kinase activation, proteases, and other factors in P-mediated calcium increase and acrosome reaction. Recent studies suggest the possibility that the sperm's response to P may be functionally related to their fertilizing ability, thus opening new perspectives in the possible development of a predictive test in the assisted reproductive techniques. PMID- 18406703 TI - Is brain tumor recurrence increased following growth hormone treatment? AB - Growth hormone (GH) deficiency and resultant growth failure are common sequelae in children with brain tumors, often requiring treatment with GH. Several studies over the past decade have addressed the concern as to whether GH treatment in these children results in an increased rate of tumor recurrence. The findings in general are reassuring, although there are analytic problems with all the studies. This present article reviews the previously published studies, as well as the experience at the Children's Hospital of Philadelphia. PMID- 18406704 TI - New frontiers in insulin receptor substrate signaling. AB - Although most tyrosine kinase growth factor receptors directly bind Src homology 2 (SH2) proteins, the insulin receptor, and a select group of other hormone receptors-including an emerging group of cytokine receptors-phosphorylate intracellular insulin receptor substrate (IRS) proteins, which subsequently bind SH2 proteins. There are currently two members of the IRS family (IRS-1 and IRS 2); these IRS proteins contain elements of substantial similarity, but may also play divergent roles in mammalian physiology. The engagement of IRS proteins by other receptors suggests that IRS proteins mediate diverse biological signals. PMID- 18406705 TI - Calcitriol-resistant rickets due to vitamin D receptor defects. AB - Calcitriol-resistant rickets (CRR) is an autosomal recessive disease due to a defect in the vitamin D receptor (VDR) or a site distal to it. The main characteristics are extreme rickets, with growth attenuation, osteomalacia, secondary hyperparathyroidism, severe dental caries, and alopecia. Serum studies reveal hypocalcemia, hypophosphatemia, very high calcitriol, and increased alkaline phosphatase levels. The clinical and chemical abnormalities do not respond to therapy with high-dose vitamin D, indicating target organ unresponsiveness. Eleven different mutations in the gene-encoding VDR have thus far been reported. They affect either the C-terminal ligand-binding region or the N-terminal DNA binding zinc-fingers sequences, with mutation hot spots identified at conserved sequences among the steroid-thyroid receptors superfamily. These result in impaired calcitriol binding to target organs, signified in vitro as failure of fibroblasts to bind [(3)H]calcitriol or to respond to calcitriol by 24 hydroxylase activity enhancement. Receptor studies and mutational analyses are used for prenatal diagnosis of CRR. Therapy with high-dose calcium overcomes the VDR defect, normalizes serum calcium, and maintains bone remodeling and mineral apposition. These responses to therapy have interesting implications upon our understanding of the potential role of calcium alone and that of vitamin D in bone physiology. Like other hormone-resistant diseases, CRR, with its various mutations, provides the opportunity for investigating the nature of vitamin D and of VDR physiology, which has been only partially explored to date. PMID- 18406706 TI - Insulinlike growth factors: Potential therapeutic options for osteoporosis. AB - The insulinlike growth factors (IGFs) are important components of the bone remodeling unit. These peptides are present in very large concentrations within the skeletal matrix and play a critical role in differentiated osteoblast function and osteoclast recruitment. IGF-I may also mediate GH's actions in the skeleton. Hence, the IGFs have promise as future anabolic agents for the treatment of osteoporosis. However, the skeletal IGF regulatory system is complex and redundant, making it exceedingly difficult to predict how administration of recombinant growth factors might ultimately affect bone mass. In addition, the lack of long-term controlled clinical trials and the prevalence of significant side effects limit the clinical application of these growth factors. Nonetheless, new and innovative delivery systems for these proteins might make the application of the IGFs a potential therapeutic option for the management of osteoporosis. PMID- 18406707 TI - The preovulatory LH surge A case of a neuroendocrine switch. AB - The preovulatory surge in LH is a unique endocrine event that involves a switch from the negative feedback effect of estrogen to a positive feedback effect. This occurs at both the level of the brain and that of the pituitary gland. Within the brain the mechanism appears to involve disinhibition of negative inputs to GnRH neurons, as well as stimulation of positive inputs. The positive feedback effects on the brain and the pituitary appear to be coordinated so that the effect of estrogen involves a number of time-delayed mechanisms. PMID- 18406708 TI - Apparent mineralocorticoid excess, 11beta hydroxysteroid dehydrogenase and aldosterone action Closing one loop, opening another. AB - The recent cloning of the human enzyme 11beta hydroxysteroid dehydrogenase type 2 (11betaHSD2), and the demonstration of point mutations or deletions in both familial and apparently sporadic cases of apparent mineralocorticoid excess (AME), underlines the importance of this enzyme in excluding glucocorticoids from mineralocorticoid receptors (MR). Although the sodium retention characteristic of AME can thus be explained by absent or very reduced (< 10%) levels of renal 11 betaHSD2 activity, whether or not the enzymatic defect contributes to the elevated blood pressure by mechanisms other than sodium retention remains to be determined. PMID- 18406709 TI - Mechanism(s) of early glucocorticoid inhibition of adrenocorticotropin secretion from anterior pituitary corticotropes. AB - Adrenal glucocorticoid hormones, released in response to stress activation of the hypothalamic-pituitary-adrenal axis (HPA), are powerful regulators of cellular function. Analysis of early (10 min to <3 h) glucocorticoid inhibition of ACTH secretion from anterior pituitary corticotropes is providing insight into potentially generic genomic mechanisms by which glucocorticoids regulate cellular excitability. Early glucocorticoid inhibition is dependent upon activation of intracellular type II glucocorticoid receptors and induction of new proteins, including the calcium-binding protein calmodulin. Glucocorticoids inhibit ACTH secretion stimulated by neuropeptide activation of the cAMP/protein kinase A (PKA) or protein kinase C (PKC) signaling pathways through mechanisms acting at, or beyond, the level of intracellular free calcium mobilization. Increasing evidence also suggests that the efficacy of early glucocorticoid inhibition is selectively modulated by the PKA pathways. The integration of molecular, electrophysiological, imaging and classic neuroendocrine techniques will further expose the molecular basis of early glucocorticoid inhibition. PMID- 18406710 TI - Molecular mechanism of cytochrome P-450-dependent aldosterone biosynthesis in the adrenal cortex. AB - In the adrenal cortex, the potent mineralocorticoid, aldosterone, is produced in the zoba glomerulosa but not in the zona fasciculata/reticularis. In rodents and humans, two distinct species of P-450(C18) (aldosterone synthase) and P 450(11beta) (11beta-hydroxylase) are expressed in the adrenal cortex. The selective expression of cytochrome P-450 species in different zones contributes to zone specificity of aldosterone synthesis. In the cow and pig, only one molecular species of P-450(11beta) having both 11beta-hydroxylase and aldosterone synthase activity is expressed throughout the adrenal cortex. P-450(11beta) in the zona fasciculata/reticularis catalyzes the formation of corticosterone but not that of aldosterone from 11-deoxycorticosterone; the same enzyme in the zona glomerulosa produces aldosterone from the same substrate, indicating that a local factor in mitochondria is likely to be involved in the selective suppression of the aldosterone synthetic activity of P-450(11beta) in the zona fasciculata/reticularis. The zone specificity of aldosterone synthesis catalyzed by P-450(11beta) in the bovine adrenal cortex appears to be due to differences in interactions between P-450(11beta) and P-450(SCC) in mitochondria in different cortical zones. Thus, two modes exist for aldosterone biosynthesis in mammals: rodent-human and bovine-porcine modes. PMID- 18406711 TI - The practical management of multiple endocrine neoplasia. AB - Adbances in the identification and localization of the abnormal genes in the multiple endocrine neoplasia syndromes have provided new methods of identifying "at risk" individuals in these families. Genetic testing using linkage analysis in multiple endocrine neoplasia (MEN) 1 and direct mutation analysis of the RET protooncogene in MEN 2 is now available for these disorders. New management issues for these disorders have resulted, and a practical approach to these issues is discussed. PMID- 18406712 TI - New aspects of the renin-angiotensin system in blood pressure regulation. AB - The renin-angiotensin system, composed of enzymatic and signal-transduction cascades, plays a key role in the regulation of arterial blood pressure and in the development of certain forms of experimental and human hypertension. The products of this system, angiotensin peptides, exert a wide range of physiologically important effects on many tissues, including those of the cardiovascular system, through their actions on angiotensin receptors. Molecular genetic and transgenic studies have begun to implicate some of the genes encoding components of the renin-angiotensin system in the development of cardiovascular diseases. Recently, we succeeded in generating mice homozygous for a targeted disruption of the angiotensinogen gene (the only known precursor of angiotensins), resulting in the complete loss of angiotensin signals in vivo. Here, we review new developments related to the functional analysis of the renin angiotensin system, in particular, by focusing on transgenic approaches including gene targeting. PMID- 18406713 TI - Molecular regulation of prostaglandin synthesis Implications for endocrine systems. AB - A wide array of prostanoids, which includes prostaglandins D(2), E(2), F(2alpha), I(2), and thromboxane A(2), has been known to exert regulatory effects in many endocrine systems for over 3 decades. More recently, however, molecular biological techniques have uncovered new findings that have brought about radical changes in our thinking about prostaglandin pharmacology and physiology. Two separate forms of cyclooxygenase (COX), a constitutive and an inducible form, have been identified. These two forms arise from separate genes whose expression is regulated differently. Moreover, genes for different receptor types and sub types of prostanoid receptors have also been cloned. The various prostanoid receptor types and subtypes are coupled to transduction systems that cause alterations in intracellular calcium and cAMP concentrations. As importantly, new sites of inhibitory action for corticosteroids and nonsteroidal antiinflammatory drugs in the COX-2 synthetic pathway have been uncovered that decrease COX-2 mRNA levels and enzyme mass. Most of the nonsteroidal antiinflammatory drugs are more effective in inhibiting activity of COX-1 compared with COX-2. This carries important clinical relevance, because COX-1 is proposed to play a role in normal physiologic processes rather than in mediating inflammation, which may explain the undesirable side effects of some of these drugs. Possible implications of these new developments on regulation of bone resorption as a representative endocrine system are considered. PMID- 18406715 TI - The N-terminal peptide of the inhibin alpha subunit What are its endocrine and paracrine roles? AB - alphaN inhibin (molecular mass 23 24 kD) is present in the pro-alphaN-alphaC subunit of inhibin and can be released by cleavage at the flanking arginine residues during posttranslational processing. Although the alphaN protein isolated from bovine follicular fluid has no inhibinlike (FSH suppressing) activity, alphaN is present in high molecular weight forms of biologically active inhibin found in follicular fluid and plasma. alphaN may modify the biological activity of inhibin by influencing its half-life or access to its receptor. alphaN may also play a role in regulating fertility through a local action on ovulation by the ovary that is independent of the actions of inhibin. The evidence suggests a unique physiological significance for the precursor peptides of the inhibin-alpha subunit in both the endocrine and paracrine control of fertility. PMID- 18406714 TI - Potential functions of plasma steroid-binding proteins. AB - The plasma steroid-binding proteins, sex hormone-binding globulin (SHBG) and corticosteroid-binding globulin (CBG), transport steroid hormones in the blood and regulate their access to target tissues. Recent biochemical and molecular analyses of these proteins and their genes, and studies of their biosynthesis and localization in the liver and other tissues during development, have led to the realization that CBG and SHBG function in much more sophisticated ways. In particular, the presence of plasma membrane binding sites for both CBG and SHBG on steroid target cells, and evidence for interactions between CBG and specific proteinases at sites of inflammation or tissue remodeling, suggest that these proteins control steroid hormone bioavailability and/or action in a highly selective or targeted fashion. This new information should not only serve to extend our understanding of the basis of steroid-hormone dependent diseases, but may influence the design of steroid hormone agonists and antagonist of therapeutic potential. PMID- 18406716 TI - Prevention of insulin-dependent diabetes-1995. AB - Despite current treatment advances, insulin-dependent diabetes mellitus (IDD) is still associated with high morbidity and mortality, and with a huge financial burden both to the individual and society. Enhanced understanding of the natural history of the prediabetic period has made the disease predictable in both higher risk nondiabetic relatives and in the general population. Investigators around the world are now collaborating on three separate multicenter, randomized, controlled trials aimed at preventing the disease in at-risk individuals, engendering cautious optimism that the days are not too far away when IDD can safely be prevented. PMID- 18406717 TI - Comparative neuroendocrinology of steroid receptor gene expression and regulation: Relationship to physiology and behavior. AB - Great diversity exists among vertebrates in reproductive behaviors and the neuroendocrine mechanisms underlying these behaviors. Comparisons of species with different hormone-brain-behavior relationships reveal three factors which may explain species differences in endocrine physiology and behavior: (a) sensitivity to sex steroid hormones, (b) hormone-dependent regulation of sex steroid hormone receptor gene expression, and (c) neuroanatomical distribution of steroid receptor gene expression, especially in nonlimbic structures. PMID- 18406718 TI - A model for farnesoid feedback control in the mevalonate pathway. AB - Mevalonate is the rate-limiting substrate leading to farnesyl pyrophosphate (FPP), the central intermediate for isoprenoids such as cholesterol, dolichols, ubiquinone, and carotenoids. One major challenge has been to identify the isoprenoid effector molecules and transcription factors mediating negative regulation in this metabolic pathway. A nuclear receptor called FXR has recently been characterized that is activated by farnesyl pyrophosphate metabolites such as farnesol, farnesal, farnesoic acid, and methyl farnesoate. FXR expression in isoprenoidogenic tissues suggests a hypothesis that these intracellular "farnesoids" may be signals for transcriptional feedback control of cholesterol biosynthesis. PMID- 18406719 TI - Adrenomedullin: A newly discovered hormone controlling fluid and electrolyte homeostasis. AB - Neural and humoral mechanisms controlling fluid and electrolyte homeostasis employ a diverse array of physiologic mechanisms that often, when aberrant, are the underlying cause of disease. Behavioral, hormonal, renal, and vascular responses to volume and osmotic challenges must be coordinated to achieve the goal of homeostasis. In recent years, it has become apparent that there exist a number of hormonal factors produced throughout the body that can coordinate these multiple regulatory mechanisms by complementary effects in several tissues. Thus, in addition to their vasoactive properties, recently characterized hormones such as the natriuretic peptides and the endothelins, as well as the better established renin-angiotensin system, exert central nervous, renal, cardiac, and pituitary effects that regulate normal fluid and electrolyte balance. Now a new player, adrenomedullin, has been added to the cast, and the interplay of multiple hormonal factors involved in the physiology and pathophysiology of volume and osmotic status continues to be elucidated. PMID- 18406720 TI - Intracellular signal transduction: The role of endosomes. AB - Polypeptide hormones, growth factors, and other biologically significant molecules are specifically internalized by target cells. Exposure of cells to these ligands results in the formation of ligand-receptor complexes on the cell surface and subsequent internalization of these complexes into the endosomal apparatus (endosomes, or ENs). The study of ENs has identified several important functions for this unique cellular organelle. These include the dissociation of ligand from receptor and receptor recycling to the cell surface and the degradation of some internalized ligands, as well as the delivery of others to lysosomes. More recently, it has become apparent that ENs fulfill another critical role, that of signal transduction. In this article, we review the evidence substantiating this role for ENs and propose three models by which ENs participate in cell signaling. PMID- 18406721 TI - Melatonin Its intracellular and genomic actions. AB - Besides its actions via membrane receptors on cells in specific areas of the brain and peripheral tissues, melatonin also has direct intracellular actions. The neurohormone melatonin is a potent free radical scavenger in vitro and a powerful antioxidant in vivo. Studies to date indicate that it is a better scavenger of the highly toxic hydroxyl and peroxyl radicals than some other known compounds. Melatonin also prevents the toxicity of singlet oxygen and stimulates the antioxidative enzyme, glutathione peroxidase. Considering its varied and potent antioxidant capability, it is possible that melatonin is an essential element of the antioxidant defense system of organisms. Besides these direct intracellular actions of melatonin, it was recently discovered that the indole also has binding sites in the nucleus of many cells. Melatonin's genomic actions are believed to follow its binding to these nuclear loci. PMID- 18406722 TI - Matrix metalloproteinases and their tissue inhibitors in endocrinology. AB - Matrix metalloproteinases (MMPs) and their natural inhibitors (TIMPs) have been widely implicated in tissue resorption, degradation, and fibrosis in a range of normal and abnormal processes, but recently defined additional actions suggest much broader and independent roles for different members of these gene families. This review examines the involvement of hormones in regulation of MMPs and TIMPs and focuses on known and potential roles of particular interest to endocrinologists. PMID- 18406723 TI - Insulin, growth factors, and cAMP: antagonism in the signal transduction pathways. AB - Depending on the cell type and the response, cAMP may either oppose or facilitate the actions of insulin and/or growth factors that signal via receptor tyrosine kinases. Recent findings indicate that the effects of the cyclic nucleotide are mediated in part by changes in the activities of important elements in the signal transduction pathways utilized by insulin and growth factors. PMID- 18406724 TI - Thyroid disease: effects on cardiovascular function. AB - Thyroid hormones exert direct effects upon the heart and indirect influences mediated via changes in cardiac work. Overt thyroid dysfunction is frequently associated with cardiovascular symptoms and signs and less frequently with significant cardiovascular morbidity due to atrial fibrillation or cardiac failure. Whether subclinical thyroid dysfunction is similarly associated with adverse effects upon the cardiovascular system remains unclear. PMID- 18406725 TI - Gonadotropin-releasing hormone neuron cell biology. AB - For those who do not work directly with this famed tissue on the underside of the brain, the hypothalamus can appear to be an intimidating network of neurons possessing a complex intercellular wiring diagram and offering a catalog of secretory products with autocrine, paracrine, and endocrine activities. For those who have been seduced into studying the multifunctional hypothalamus, especially its central role in reproductive biology, things recently have gotten a whole lot better. PMID- 18406726 TI - Pulsatile gonadotropin-releasing hormone release and its regulation. AB - Several proposals have been put forward to explain the pulsatile pattern of gonadotropin-releasing hormone (GnRH) secretion form the hypothalamus, and the receptor-mediated modulation of such pulsatility. The mechanisms underlying these phenomena are not well defined, but the findings of intrinsic rhythmic activity of GnRH neurons, and the presence of autocrine GnRH action therein, have identified important elements in the episodic mode of neuropeptide release. PMID- 18406727 TI - GnRH gene expression in neuronal cell lines. AB - Little is known about the mechanisms that subserve GnRH synthesis. The recent availability of clonal populations of GnRH neurons enables us for the first time to examine the structural organization of the GnRH locus. Studies are underway to dissect the manner in which neural specific expression and developmental regulation of GnRH are programmed. These studies will complement and provide direction to future animal studies designed to better understand the control of GnRH gene expression. PMID- 18406728 TI - Regulation of gonadotropin-releasing hormone neurons by basic fibroblast growth factor. AB - The development of a functional network of gonadotropin-releasing hormone (GnRH) neurons in the central nervous system requires a series of complex regulatory mechanisms, presumably mediated in part by neurotrophic factors. The difficulty in studying factors regulating the development of GnRH neurons stems from their paucity and scattered distribution in the brain; as a result, little was known about the role of neurotrophic factors in the development of the mature GnRH neuronal network. Recent utilization of immortalized GnRH neuronal cell lines (GT1) has enabled us to identify and study specific neurotrophic factors and their functions in vitro. The potent neurotrophic effect of basic fibroblast growth factor (bFGF) and the presence of a high abundance of receptors for bFGF in GT1 cells have led to the hypothesis that bFGF may be an important regulator of GnRH neuron expansion, survival, migration, and connectivity. PMID- 18406729 TI - Protein phosphorylation, kinases, and phosphates: 20th European Symposium on Hormones and Cell Regulation Strasbourg, France, September 22-25, 1995. PMID- 18406730 TI - Androgens as anabolic agents for bone. AB - Androgens are classically considered anabolic agents, and in fact there is abundant evidence in many tissues that corroborates strong positive effects of androgens on proliferation and growth. In bone as well, there is clear evidence that androgen action is associated with an increase in skeletal mass, particularly during growth. Whether androgens can be considered anabolic in the skeleton later in life (when therapeutic increases in bone mass are of most clinical interest) is still a matter of some debate. PMID- 18406731 TI - Insulinlike growth factor I and the kidney. AB - Insulinlike growth factor I (IGF-I), IGF-I receptors, and IGF-binding proteins are expressed in different segments of the nephron in a relationship that suggests autocrine, paracrine, and endocrine modes of action. IGF-I contributes to compensatory nephron growth in a variety of experimental renal diseases with loss in functioning nephron number, and to tissue repair after ischemic acute tubular necrosis. IGF-I causes arteriolar dilatation in the kidney and increases the glomerular filtration rate in experimental animals, in normal subjects, as well as in patients with chronic renal failure, and this effect of the peptide is probably mediated by nitric oxide. IGF-I raises proximal tubular phosphate reabsorption and may increase sodium absorption in distal tubules. In the nephrotic syndrome, IGF-I- and IGF-binding protein complexes are excreted in urine and IGFBP-3 protease activity is increased, causing complex abnormalities in the IGF-system. PMID- 18406732 TI - The thyrotropin-releasing hormone gene: differential regulation, expression, and function in hypothalamus and two unexpected extrahypothalamic loci, the heart and testis. AB - Thyrotropin-releasing hormone (TRH) is the key regulator of the synthesis and secretion of TSH in animals and humans (Wilber and Yamada 1990). The biological implications of this peptide, the first releasing hormone to be characterized, has generated a large literature regarding both pituitary TSH and extrapituitary roles of TRH as a neurotransmitter and/or neuro-modulator in the central nervous system (O'Leary and O'Connor 1995, Morley 1981). In this review, new areas of TRH biology are explored, focused on the differential regulation of the TRH gene by triiodothyronine (T(3)) and other substances in the hypothalamus and two unexpected extrahypothalamic loci, the heart and testis. These new directions should enlarge our understanding concerning how hormones like T(3) regulate genes negatively and selectively with the identical receptors and DNA elements required for positive gene stimulation. In addition, regulatory studies of the TRH gene by T(3) should be relevant to other hormone receptor interactions with DNA sequences in general, as glucocorticoids, mineralocorticoids, sex steroids, vitamin D, and retinoic acid are ligands for homologous receptor proteins in the nuclear receptor superfamily. PMID- 18406733 TI - Converting heterodimeric gonadotropins to genetically linked single chains: new approaches to structure activity relationships and analogue design. AB - One of the distinguishing features of the gonadotropin and thyrotropin hormone family is their heterodimeric structure; the subunits combine early in the secretory pathway and only the dimers are capable of binding to receptors. Therefore, assembly is rate limiting in the production of functional heterodimers, a problem encountered when removing the carbohydrates from one or both subunits as discussed in this review. If the heterodimers can be expressed as single chains, this might avoid mutagenesis-induced defects in secretion and combination of individual subunits for structure-function studies and analogue design. Here we discuss the feasibility of this approach for such problems. PMID- 18406734 TI - New light is shining on the melatonin rhythm enzyme: the first postcloning view. AB - One of the most interesting molecules in circadian biology is serotonin N acetyltransferase (arylalkyfamine N-acetyltransferase, AANAT), the enzyme that controls the daily rhythm in pineal melatonin production and blood melatonin. The recent cloning of AANAT cDNA has led to the characterization of the human gene; the realization that AANAT represents a unique gene family; the discovery of circadian rhythms in AANAT mRNA; the determination of the basis of transsynaptic and cellular regulation of expression of the AANAT gene; a new understanding of the relationship of AANAT mRNA and activity; and the surprising finding of strong expression of the AANAT gene in the retina and significant levels in select brain regions, the pituitary gland, and testes. The cloning of AANAT cDNA has not only made it possible to answer longstanding questions in circadian biology, but has also raised stimulating new issues. PMID- 18406735 TI - Preclinical Cushing's syndrome. PMID- 18406737 TI - 17beta-Hydroxysteroid dehydrogenase 3 deficiency. AB - Five known isoenzymes catalyze the 17beta-hydroxysteroid dehydrogenase reaction that controls the interconversion of estrone and estradiol and of testosterone and androstenedione. Mutations in the 17beta-hydroxysteroid dehydrogenase 3 gene impair the formation of testosterone in the fetal testis and give rise to genetic males with normal male Wolffian duct structures but female external genitalia. Such individuals are usually raised as females but virilize at the time of puberty as the result of a rise in serum testosterone. The 14 mutations characterized to date in 17 affected families include 10 missense mutations, 3 splice junction abnormalities, and 1 frame shift mutation. Three of the mutations have occurred in more than 1 family. The usual mechanism for testosterone formation in affected individuals at puberty appears to be conversion of androstenedione to testosterone in extraglandular tissues by one or more of the unaffected 17beta-hydroxysteroid dehydrogenase isoenzymes. PMID- 18406738 TI - Neuroendocrinology of the mother-child interaction. AB - Mother-child interaction is usually described in psychological terms; however, it is becoming increasingly clear that neuroendocrine mechanisms are involved in the mother-child interaction. Mother and fetus influence each other by hormonal mechanisms in utero, and after birth, mother and child interact by way of sensory stimulation. In the breastfeeding situation, the infant's suckling stimulates maternal oxytocin and PRL release and, thereby, milk production and release. In addition, oxytocin, by way of circulating and central neurogenic oxytocinergic mechanisms, adapts maternal physiology and endocrinology to allow milk production. At the same time, suckling influences the physiology and endocrinology of the infant to optimize the growth process. It seems that tactile, olfactory, visual, and perhaps other types of sensory stimuli contribute to the adaptive changes in both mother and infant. In addition, behavioral changes are induced in the mother in connection with birth and breastfeeding. Data are accumulating that suggest that the attachment between mother and infant is facilitated by skin-to-skin contact early postpartum. Furthermore, calm and social interactive behavior is promoted in the mother. Central oxytocinergic mechanisms activated in connection with birth and breastfeeding seem to be involved in the behavioral changes. PMID- 18406739 TI - Role of transcription factors CREB and CREM in cAMP-regulated transcription during spermatogenesis. AB - The cAMP response element binding protein (CREB) and the cAMP-responsive element modulator (CREM) are cyclically expressed at high levels during spermatogenesis. Cyclical expression of CREB and CREM in germ and somatic Sertoli cells correlates with the fluctuations in cAMP signaling induced by the pituitary gonadotropic hormones FSH and LH both during sexual maturation of the testis and during the 12 day cycles of spermatogenesis that occur in the adult testis. CREB and CREM are expressed at different times during the spermatogenic cycle, undergo programmed sequential switches from activator to repressor isoforms by mechanisms of alternative exon splicing and promoter usage, and are autoregulated by cAMP signaling in opposing directions. cAMP response elements located in the promoter of the CREB gene upregulate the expression of activator CREBs, whereas cAMP autoregulatory response elements in the internal promoter of the CREM gene induce expression of repressor CREM isoforms. The complex mechanisms for the regulation of the expression of CREB and CREM in the testis appear to reflect critical adaptations for regulating key target genes essential for the development of germ cells. PMID- 18406740 TI - Viral elements in autoimmunity of type I diabetes. AB - The involvement of viruses in the etiology of insulin-dependent diabetes was hypothesized more than 15 years ago based on solid scientific evidence, however; the true existence and the real nature of this involvement still eludes our efforts. This may be due to the frequently long interval between viral exposure and diabetes onset that makes the direct cause-effect relationship difficult to prove. PMID- 18406741 TI - Growth hormone and aging: Regulation, signal transduction and replacement therapy. AB - It is widely accepted that during the aging process a number of alterations occur at the molecular, cellular, and tissue levels, ranging from an alteration in receptor signal transduction and gene expression to structural and morphological changes in various tissues. There is abundant empirical and scientific evidence to support the hypothesis that many of these aging processes are closely related to a decline in hormone concentrations and/or hormone action, but the etiology of these deficiencies remains elusive. GH and insulinlike growth factor-I (IGF-I) are two potent anabolic hormones that decrease with age and appear to contribute to the loss of tissue function that is associated with normal aging. In this review, age-related changes in the regulation of these hormones are detailed as well as relatively new information on mechanisms of tissue resistance to GH. Finally, the potential use of GH as a therapeutic intervention to delay physiological changes associated with age is discussed. PMID- 18406742 TI - Jansen's metaphyseal chondrodysplasia: a disorder due to a PTH/PTHrP receptor gene mutation. AB - Jansen's metaphyseal chondrodysplasia (JMC) is a rare genetic disorder that is characterized by short-limbed dwarfism and severe, agonist-independent hypercalcemia. An activating PTH/PTHrP receptor mutation that results in constitutive cAMP accumulation was recently identified in the genomic DNA of a patient with this disorder. These findings provide a plausible explanation for the abnormal regulation of growth-plate chondrocytes and mineral ion homeostasis in JMC, and may have significant implications for understanding the broader biological role of PTHrP and its receptor. PMID- 18406743 TI - Recent advances in parathyroid imaging. AB - Although no existing imaging procedure is as effective as an experienced surgeon for locating abnormal parathyroid glands in patients without previous neck surgery, preoperative parathyroid localization is considered essential for patients undergoing reoperations. The need for parathyroid imaging in patients undergoing an initial exploration remains controversial. Scintigraphy with (99m)Tc-sestamibi has largely replaced (99m)Tc-pertechnetate/(201)Tl chloride subtraction scintigraphy for parathyroid imaging because of its superior sensitivity and false-positive rate. Positron emission tomography, another technique recently applied to parathyroid imaging, is of uncertain value at present. PMID- 18406744 TI - GnRH and PACAP action in gonadotropes: cross-talk between phosphoinositidase C and adenylyl cyclase mediated signaling pathways. AB - In order to respond appropriately to their environment, gonadotropes, like other cells, must integrate informational input from multiple ligands acting through multiple intracellular signaling pathways. In recent years, an increasing number of examples of functional interactions between the phosphoinositidase C (PIC) and adenylyl cyclase signaling pathways in gonadotropes have been described, and the discovery that these cells are targets for pituitary adenylyl cyclase activating peptide (PACAP) has provided a physiological context for earlier work on gonadotrope regulation by cAMP. It has become clear that gonadotropes possess multiple PIC-coupled receptor types, in addition to receptors activating adenylyl and guanylyl cyclases, so that the potential for both coincidence signaling and cross-talk in these cells is immense; examples of both are seen in the effects of PACAP and GnRH on Ca(2+) mobilization and adenylyl cyclase activation in alphaT3 1 cells. In these cells, GnRH, acting via PIC-coupled receptors, can dramatically inhibit adenylyl cyclase activated by PACAP, but can also alter cellular levels of protein kinase A subunits, providing a mechanism for coordinated regulation of both messenger and effector. PMID- 18406745 TI - Steroid receptors and action in the primate follicle. AB - Studies localizing steroid receptor (R) proteins and their mRNAs to the primate ovary are consistent with a local role for progesterone (P) and androgen (A) in modulating follicle growth and/or maturation via classic R-mediated pathways, but data in support of estrogen (E) action remain equivocal. Investigations of the hypothesis that steroids play a pivotal role in folliculogenesis or gametogenesis in macaques receiving inhibitors of steroid enzyme synthesis and in women with congenital steroid enzyme deficiencies reveal that (a) antral follicle growth, maturation, and luteinization, as well as oocyte meiosis, do not require high or increasing E levels; (b) elevated A to E ratios are detrimental to the gametogenic functions of the primate follicle, but do not alter growth and maturation of antral follicles; and (c) ovulation and luteinization of the mature follicle are dependent on local P actions. The genomic actions of steroids likely vary between compartments and with the developmental state of the follicle; however, potential nongenomic actions of steroids in follicles remain largely undefined. Future advances will rely on identification of discrete biochemical, morphological, and functional correlates of steroid hormone action in the somatic and gametogenic compartments of the primate follicle throughout its life cycle. PMID- 18406746 TI - Premenstrual syndrome. AB - Within the past decade, premenstrual syndrome (PMS) has become the subject of rigorous scientific scrutiny. As a result, diagnostic criteria have been developed, and the pathophysiology of the disorder has been partially elucidated. The preponderance of evidence suggests that the disorder is the result of the interaction of cyclic changes in estrogen and progesterone with specific neurotransmitters. Serotonin and gamma-amino butyric acid (GABA) appear to be especially important in this regard. Increased understanding of PMS has enabled the development of specific treatment modalities that, unlike previous prescriptions, have demonstrated efficacy in rigorous and reproducible studies. PMID- 18406747 TI - The inhibin/activin family and ovarian cancer. AB - Ovarian cancer is the most common fatal malignancy of the female reproductive tract and frequently presents at an advanced stage. There is, thus, a great need for more sensitive and accurate methods of diagnosis, including better tumor markers. A rapidly emerging technique is the application of assays for the inhibin peptide family in patients with various forms of ovarian cancer. Currently, in assays that detect molecules containing the inhibin a subunit, more than 80% of postmenopausal patients with mucinous epithelial ovarian tumors, and virtually all with granulosa-cell tumors, have elevated inhibin levels. The detection of activin in some tumor tissues suggests that this peptide may also prove to be of interest once satisfactory and specific assays for the circulating peptide are available. It is likely that significant progress will be made in this field within the next 5 years. PMID- 18406748 TI - The roles of the nuclear receptor steroidogenic factor 1 in endocrine differentiation and development. AB - The orphan nuclear receptor steroidogenic factor 1 (SF-1) has emerged as a critical determinant of adrenal and gonadal differentiation, development, and function. SF-1 was initially isolated as a positive regulator of the cytochrome P450 steroid hydroxylases in the adrenal glands and gonads; developmental analyses subsequently showed that SF-1 was also expressed in the diencephalon and anterior pituitary, suggesting additional roles in endocrine function. Analyses of knockout mice deficient in SF-1 revealed multiple abnormalities, including adrenal and gonadal agenesis, male to female sex reversal of the internal genitalia, impaired gonadotrope function, and absence of the ventromedial hypothalamic nucleus. Taken together, these results implicate SF-1 as a global regulator within the hypothalamic-pituitary-gonadal axis and the adrenal cortex. PMID- 18406749 TI - Seasonal diminution of vitamin D stores in the United States. Can darker winters lead to lighter bones? AB - Recent reports indicate that many older Americans have inadequate vitamin D sources and little to no seasonal increase in their body stores. The attendant secondary hyperparathyroidism of incipient and subtle osteomalacia could weaken cortical bone and aggravate the osteoporotic process linked to aging. Increased awareness, by physicians and our elderly, of the importance of avoiding mild, seasonal or perennial vitamin D deficiency could lead to better implementation of vitamin D supplementation schedules. PMID- 18406750 TI - Cushing's disease in childhood. AB - Cushing's disease is rare in childhood. There is an equal sex incidence, and it accounts for approximately 75% of pediatric causes of Cushing's syndrome. Predominant features are weight gain, growth failure, virilization, and headache. Following confirmation of the presence of inappropriate hypercortisolemia, the accurate differential diagnosis to establish the pituitary as the source of excessive ACTH secretion involves demonstration of > 50% suppression of circulating cortisol during high-dose dexamathasone administration and exaggeration of the cortisol response to corticotropin-releasing hormone (CRH). Imaging of the pituitary reveals a microadenoma in only a minority of cases, but inferior petrosal sinus sampling for ACTH can be of value in confirming the pituitary location of the tumor and possibly its lateralization. Primary therapy is transsphenoidal surgery, which can be supported by direct pituitary irradiation if hypercortisolemia persists. In experienced hands, the therapeutic outcome is good. PMID- 18406751 TI - Combination therapy with oral antihyperglycemic agents in the treatment of non insulin-dependent diabetes mellitus. AB - Combination therapy, using drugs from different classes of oral hypoglycemic agents, offers several advantages in the treatment of non-insulin-dependent diabetes mellitus (NIDDM). Appropriate combinations can significantly improve glycemia and often obviate insulin treatment. In general, no increased toxicity or significant interactions are observed. PMID- 18406752 TI - StAR-A tissue specific acute mediator of steroidogenesis. AB - The rate-limiting and acutely regulated step in steroid hormone biosynthesis is the translocation of cholesterol, the precursor of all steroid hormones, from the mitochondrial outer membrane to the inner membrane, where it is converted to pregnenolone by the cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc). This step has long been known to be dependent upon the de novo synthesis of a labile protein factor, which is required for the intramitochondrial translocation of cholesterol. Recently, the Steroidogenic Acute Regulatory (StAR) protein has been shown to have an indispensable role in acute steroid production and is proposed to be this labile protein factor. Given the fundamental importance of StAR as a key regulator of steroid hormone biosynthesis, the next frontier for researchers is elucidating the molecular mechanisms that control StAR expression and function. PMID- 18406753 TI - vgf A neurotrophin-inducible gene expressed in neuroendocrine tissues. AB - vgf is an inducible gene, highly sensitive to nerve growth factor (NGF) and remarkably upregulated in the "early-delayed" phase of response (within a few hours). It encodes a 617-amino acid polypeptide (VGF protein) bearing no significant homology with known sequences and restricted to certain peptide/amine producing endocrine cells, and neurons (for example, adenohypophysial and adrenal medullary cells, or hypothalamic neuroendocrine neurons). VGF is stored and transported in secretory granules and processed to intermediate-small molecular weight products, which are preferentially released. Striking changes in both VGF mRNA and immunolocalization are found in physiological conditions (for example, estrous cycle) and in experimental models of stimulation affecting hypothalamic and other neurons. Functional roles of VGF are to be sought in secretory granule formation and regulation, and/or in the production of potentially bioactive peptides. PMID- 18406754 TI - Regulation of the hypothalamo-pituitary-adrenocortical axis in fetal sheep. AB - Development of the fetal hypothalamo-pituitary-adrenal (HPA) axis is required for normal fetal life and subsequent neonatal health. Activation of the fetal pituitary gland results in the synthesis and release of glucocorticoids from the adrenal cortex. Glucocorticoids promote maturation of several organ systems, are important in responses of the fetus to stress, and are involved in the initiation of parturition in several species. The expression of hypothalamic and pituitary genes associated with HPA function is apparent early in gestation in fetal sheep, although the endocrine changes associated with maturation and parturition do not occur until the last fifth of gestation. In this connection, the fetal HPA axis can be activated by treatment with hypophysiotrophic factors or moderate stress throughout gestation. This review focuses on the development of neuroendocrine mechanisms controlling HPA function during fetal life. PMID- 18406755 TI - Thyroid-specific enhancer-binding protein Role in thyroid function and organogenesis. AB - Thyroid-specific enhancer-binding protein (T/EBP), also known as thyroid-specific transcription factor-1 (TTF-1), is a trans-activating transcription factor known to be involved in the tissue-specific expression of genes encoding thyroglobulin (TG), thyroid peroxidase (TPO), and thyrotropin receptor (TSHR) in the thyroid, and surfactant proteins A, B, and C, and Clara cell secretory protein in the lung. T/EBP is a homeodomain-containing DNA-binding protein found in the thyroid, lung epithelium, and specific areas of the forebrain during early embryogenesis. A mutant mouse lacking T/EBP expression was recently generated that is missing the thyroid, lung, ventral forebrain, and pituitary. These results clearly establish that T/EBP functions not only as a trans-activating factor to regulate expression of genes in the thyroid and lung, but also is required for the organogenesis of the thyroid, lung, ventral forebrain, and pituitary. PMID- 18406756 TI - Immunotherapies in diabetes. AB - Insulin-dependent diabetes (IDD) is a serious, life-long disease replete with life-threatening complications that are not preventable through conventional insulin replacement therapies. The prolonged prodromal period of autoimmunity to beta cell antigens offers multiple intervention opportunities. These can target different steps that precede final destruction of insulin-secreting beta cells and clinical onset of the disease. All current and proposed immunotherapies are experimental procedures that have proven to be protective in animal models, especially the nonobese diabetic (NOD) mouse. This brief review deals with a selected list of nonspecific and autoantigen-specific immunotherapies that may bring hope in the near future to individuals at risk of developing the disease. None are yet proven to be effective in humans. PMID- 18406757 TI - Paget's disease of bone possible viral basis. AB - In 1974, the first report of the presence of viral-like inclusions in the osteoclasts of patients with Paget's disease appeared (Rebel et al 1974). This observation, first made in France, was followed by confirmatory reports from North America, Europe, and Asia. Although no more compelling hypothesis than a viral etiology has been proposed subsequently, definitive proof of viral etiology is still lacking. This article summarizes the research of the past 22 years, which has been directed at defining the exact nature of the viral-like inclusions that are invariably found in the osteoclasts of Paget's disease. PMID- 18406758 TI - New perspectives in polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is, perhaps, the most common endocrinopathy affecting premenopausal women. Of such women, 5%-10% have the classic endocrine syndrome of hyperandrogenism and chronic anovulation. The syndrome not only has these long-appreciated reproductive morbidities, but it also has more recently recognized important metabolic consequences related to insulin resistance. This article reviews the current state of the field with respect to the pathogenesis of PCOS and the insulin resistance associated with it. PMID- 18406759 TI - Gonadal hormone feedback on pituitary gonadotropin genes. AB - The pituitary gonadotropins LH and FSH, which act on the ovaries and testes to promote gametogenesis and sex steroid production, are regulated by changes in the levels of steroids and gonadal peptides. Steroid feedback can be positive, as demonstrated by the estrogen and LH surge at ovulation, or negative, as demonstrated by the rise in LH and FSH after gonadectomy or reductions in steroid synthesis. Modulatory effects of the steroids estrogen and testosterone may be mediated directly at the level of the pituitary cells, or by alterations in the release of the hypothalamic releasing factor GnRH. Gonadal peptides, including activin and inhibin, have been shown to have direct effects on pituitary cells to alter FSH synthesis specifically, with no effects on LH. Changes in gonadotropin subunit gene transcription and mRNA levels occur very rapidly and have profound effects on physiological levels of the hormones. In this article, the effects of the gonadal steroids and peptides as modifiers of the rat gonadotropin genes in a subunit specific manner are reviewed, and the physiological implications discussed. PMID- 18406760 TI - Structure-function studies of human TSH: new advances in design of glycoprotein hormone analogs. AB - Recent progress in structure-function studies of glycoprotein hormones has provided new insights into the molecular mechanisms of action of these hormones and has further supported the concept that physiological modulation of assembly, bioactivity, and clearance of these hormones is dependent on specific structural components. This review emphasizes current advances in the structure-function relationships of human TSH, which have contributed to further elucidation of common and hormone specific features within the glycoprotein hormones family. Novel strategies are now being applied to investigate the role of individual structural elements. The principks discovered in such studies are essential to understand the physiological regulation of hormone bioactivity and allow for the rational design of novel analogs with potential therapeutic applications. PMID- 18406761 TI - Growth Hormone therapy for adult Growth Hormone deficiency. AB - GH deficiency in adult life is associated with a number of adverse biological changes including osteopenia, reduced exercise capacity, altered body composition, deleterious alterations in the lipid profile and insulin status, and reduced quality of life. Potentially, most of these changes can be reversed by GH replacement therapy. In an era of health rationing, however, GH replacement is unlikely to be offered to every GH-deficient adult. Therefore, we have proposed a strategy aimed at delineating which adults with GH deficiency might benefit most from GH therapy. PMID- 18406762 TI - Acquired estrogen independence and antiestrogen resistance in breast cancer: estrogen receptor driven phenotypes? AB - Endocrine-responsive breast tumors appear to follow a predictable pattern of progression from estrogen dependence to estrogen independence, ultimately leading to a phenotype characterized by crossresistance among all endocrine therapies. Cells acquiring a multihormone-resistant phenotype, however, frequently retain expression of the cellular receptors for estrogen (ER) and progesterone (PgR). The proliferation of some of these tumors may be driven by a ligand-independent activation of the remaining steroid hormone receptors. Several intracellular secondary messenger systems can potentially activate ER through altering its pattern of phosphorylation in the absence of estrogens. Emerging evidence suggests that, for many estrogen-regulated genes, both the promoter and cellular contexts are critical factors in regulating their transcription by ER. The cellular context may be important because of the presence/absence of several coregulators of ER function, and this context may be unstable in tumor and some normal cell populations. Thus, the pattern of genes regulated by the transcriptional activities of the ER also may change with time, facilitating the emergence of divergent endocrine-responsive phenotypes. It is this pattern of regulation that may be important for conferring each specific phenotype. The repression or induction of the functionally relevant genes responsible for conferring each of the phenotypic changes represents an estrogen-regulated gene network. These networks will contain genes that are regulated, both directly and indirectly, by the activation of ER. Several growth-regulatory gene networks may exist concurrently, providing a cell with several interrelated pathways for controlling its proliferation. The identity of those estrogen-regulated genes that are responsible, for regulating proliferation remains unknown. PMID- 18406763 TI - Galanin: an unassuming neuropeptide moves to center stage in reproduction. AB - Galanin is a neuropeptide widely distributed throughout the reproductive system, including the brain, pituitary, testes, ovaries, and accessory reproductive tracts of both sexes. Within the hypothalamus, a subset of GnRH-producing neurons also synthesizes galanin. Galanin gene expression in these GnRH neurons is highly inducible by estrogen. Galanin is thought to play an important role in the generation of the preovulatory LH surge, and may serve other functions yet to be discovered. PMID- 18406764 TI - Lessons from transgenic and knockout animals about noninsulin-dependent diabetes mellitus. AB - The application of transgenic techniques to alter gene expression in vivo has provided new models to evaluate the role of specific genes in the complex pathogenesis of noninsulin-dependent diabetes mellitus (NIDDM). In this review, we summarize methods used to create transgenic animals and highlight results from those models which have contributed to our understanding of the overall pathophysiology of NIDDM. Transgenic animal models have clearly demonstrated the requirement for normal insulin action in skeletal muscle, adipose tissue, and liver, as well as normal insulin secretion by the pancreatic beta-cell, in the maintenance of glucose homeostasis. In addition, these data confirm that isolated defects in single critical genes, including the insulin receptor, IRS-1, and glucokinase, may play a role in the development of some types of insulin resistance and NIDDM. However, it is likely that multiple additive defects, both genetic and acquired, are required to produce the full clinical syndrome typical of more common forms of NIDDM. PMID- 18406765 TI - Pathogenesis of diabetic nephropathy. AB - Diabetic nephropathy is the most important long-term complication of diabetes mellitus and is a major cause of end-stage renal disease. The condition is associated with excess cardiovascular morbidity and mortality, as well as other diabetic microvascular complications. The etiopathogenesis of diabetic nephropathy appears to involve both genetic and environmental factors leading to disease in a subgroup of patients. Improved knowledge of the natural history and pathophysiology of the condition have enabled therapeutic strategies to be employed that have improved the outlook for patients with nephropathy. Strategies aimed at the identification of patients at high risk of nephropathy could have a major impact on the overall cardiovascular and renal morbidity in diabetic patients. PMID- 18406766 TI - Steroid regulation of sexual behavior. AB - Investigation into the hormonal control of sexual behavior has a rich and extensive history. For many researchers currently active in the field, the physiological psychologist Frank A. Beach is recognized as the modern father of the study of hormones and behavior. His publication of the seminal book Hormones and Behavior-A Survey of Interrelationships Between Endocrine Secretions and Patterns of Overt Response, published in 1948, was a compilation of the previous 20 years of research establishing that gonadal secretions acted in the brain and modulated behavior. The question of precisely how hormones can alter brain functioning in a coordinated fashion and profoundly influence the patterns of behavioral responsiveness remains unanswered. As with many research areas, application of new techniques and approaches to the problem reveals additional layers of complexity and previously unimagined relationships between hormones, brain, and behavior. In addition, with the increasing understanding that the brain is a target organ for steroids, the implications of the ramifications of this steroid sensitivity have broadened. The hormonal regulation of sexual behavior is not an isolated aspect of steroid action in the brain; rather, it is one component of a host of physiological responses influenced by steroids. These include such diverse responses as anxiety, aggression, feeding, and learning and memory. An appreciation of the diverse effects of steroids has emerged from studies on sexual behavior, and a mutually beneficial relationship between this and other aspects of behavioral neuroscience has flourished and endured. As with all of neuroscience, this research area has been dynamic and progressive and has additionally benefited from a long history of comparative and integrative approaches to animal behavior. PMID- 18406767 TI - Steroid modulation of neurotransmitter function to alter female reproductive behavior. AB - The hormonal induction of reproductive behavior in rodents is well established, but many of the mechanisms mediating this induction remain unknown. The ability of the gonadal steroids, estrogen and progesterone, to act simultaneously upon several neurotransmitter systems and at multiple loci in a coherent manner helps to coordinate the timing of behavioral receptivity with ovulation and assures maximal reproductive success. Investigations revealing the multitude of steroid actions on neurotransmitters illustrate the neuronal plasticity present in the adult brain. Furthering our understanding of the mechanisms by which steroids alter brain functioning to change behavior profoundly, but transiently, serves to provide information not only in the field of reproductive biology but that of basic neuroscience as well. PMID- 18406768 TI - Neural aromatization and the control of sexual behavior. AB - Neural aromatization of androgens to estrogens is known to be a critical step in the development and adult expression of male sexual behavior in a variety of species. The medial preoptic area (mPOA) is an important site of aromatization necessary for the expression of copulatory behavior in males. The neuroendocrine regulation of female sexual behavior in the musk shrew, an evolutionary "primitive" insectivore, shares several similarities with the regulation of male sexual behavior in many other species. We review the evidence that neural aromatization in the mPOA triggers female sexual behavior in the musk shrew, and speculate on the presence of a similar pathway in other mammalian species. PMID- 18406769 TI - Functions of the steroid-responsive neural network in the control of male hamster sexual behavior. AB - Gonadal steroid receptor-containing neurons in the brain are densely interconnected to form a steroid-responsive neural network within the limbic system. The possible functions of such a network include redundancy, signal amplification, stability, and selective filtering of hormonal cues to control steroid-dependent aspects of neuroendocrine secretion and behavior. Recently, the neural circuitry underlying male sexual behavior in the Syrian hamster has been used as a model for testing certain of these concepts. These studies provide functional evidence to support the network properties of gonadal steroid responsive neurons in controlling hormone-dependent sexual behavior. PMID- 18406770 TI - Androgens and male sexual behavior from mice to men. AB - Exogenous androgenic hormones or drugs that affect endogenous androgen levels are now used in a variety of human applications [cf. Bardin et al. (1991)]. Therefore, an increasing number of well-controlled physiological, clinical, and psychological studies have examined the effects of androgens on muscle, prostate, lipids, and bone, as well as the cardiovascular, immune, and nervous systems. The increasing number of androgens and delivery systems that are currently available or under development should also increase the prevalence of androgen therapy (Table 1). As androgen use increases, a crucial area of research is the effect of androgens on sexual behavior, which encompasses sexual desire, motivation, performance, and satisfaction. There is a large body of animal data documenting hormonal regulation of male sexual behavior and the neural sites of action of these hormones. Clinical data on the effects of androgens on human male sexuality come from correlative studies of endogenous testosterone levels and sexual function, treatment of men who are testosterone deficient, and men using hormonal contraceptives. PMID- 18406771 TI - Introduction. PMID- 18406772 TI - Why are endocrine pituitary cells excitable? AB - Since 1975, endocrine pituitary cells have been known to be excitable neuronlike cells. Using powerful single-cell approaches, in particular the patch clamp electrophysiological recording technique and the monitoring of Ca(2+) with fluorescent probes, solid evidence has been provided in the last 10 years that intracellular Ca(2+) signals are produced by stimulators and inhibitors of secretion via the modulation of action potentials in isolated pituitary cells. As cytosolic Ca(2+) changes are thought to control numerous cellular functions (for example, secretion, protein synthesis, gene expression, and proliferation) over a long time scale-milliseconds to hours-it is now time to address the long-standing question of what functions would be physiologically controlled by electrical excitability in intact pituitary tissue. PMID- 18406773 TI - Corticotropin-releasing hormone and calcium signaling in corticotropes. AB - Corticotropin-releasing hormone (CRH) stimulates ACTH secretion from anterior pituitary corticotropes, largely, but possibly not exclusively, via activation of the adenylyl cyclase cascade. CRH stimulates secretion by increasing Ca(2+) influx and by Ca(2+)-independent mechanisms. As Ca(2+) influx is largely regulated by membrane electrical properties, we review the effects of CRH on membrane excitability and changes in cytosolic Ca(2+). We also speculate on possible pathways for CRH modulation of exocytosis by Ca(2+) independent mechanisms. PMID- 18406774 TI - Characterization of the calcium response to thyrotropin-releasing hormone in lactotrophs and GH cells. AB - Thyrotropin-releasing hormone (TRH) acts via a G-protein-coupled receptor on lactotrophs to increase the intracellular free calcium ion concentration, [Ca(2+)](i). The [Ca(2+)](i) response depends on both TRH concentration and the duration of TRH exposure. An initial, short-lived [Ca(2+)](i) spike results from release of Ca(2+) from intracellular stores, whereas a later sustained [Ca(2+)](i) increase, often characterized by [Ca(2+)](i) oscillations, results from an influx of extracellular Ca(2+) through both voltage-gated and non-voltage gated, store-operated Ca(2+) channels. The initial spike phase predominates at high doses of TRH, whereas the plateau phase predominates at low doses. The mechanisms underlying the complex [Ca(2+)](i) response to TRH are discussed. PMID- 18406775 TI - Pituitary adenylate cyclase-activating polypeptide regulates [Ca(2+)](i) and electrical activity in pituitary cells through cell type-specific mechanisms. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a recently identified hypothalamic factor that acts on a variety of anterior pituitary cell types. It is clear, however, that its actions are not mediated by the same intracellular signaling mechanisms in each cell type. The signaling pathways by which PACAP regulates changes in [Ca(2+)], and electrical activity in rat somatotrophs and gonadotrophs is described in the present article. Finally, the possibility that the differences in PACAP-regulated signaling in anterior pituitary cells is due to the differential expression and coupling of PACAP receptor subtypes is discussed. PMID- 18406776 TI - GnRH-induced calcium and current oscillations in gonadotrophs. AB - The rat pituitary gonadotroph is a well-studied cell model for investigation of the oscillatory nature of calcium signaling in agonist-stimulated excitable cells. Cytosolic calcium levels ([Ca(2+)](i)) in gonadotrophs are controlled by two distinct oscillators, a plasma membrane oscillator that generates extracellular calcium-dependent low-amplitude [Ca(2+)](i) spiking in unstimulated cells and an endoplasmic reticulum oscillator that is activated by calcium mobilizing receptors for GnRH, endothelin, and pituitary adenylate cyclase activating polypeptide. In this review, the characteristics of the spontaneous and agonist-induced calcium oscillations in gonadotrophs and the coordinate actions of the two oscillators during GnRH action discussed. PMID- 18406777 TI - Calcium signaling and secretion in pituitary cells. AB - An important trigger of hormone secretion from pituitary cells is a rise in cytosolic Ca(2+) ([Ca(2+)](i)). Pituitary cells may modulate [Ca(2+)](i) by an increased membrane flux from the extracellular space and/or by a release from intracellular stores. Both mechanisms can support exocytosis, although in different pituitary cell types one or the other mechanism may predominate. Molecular events transducing a rise in [Ca(2+)](i) into hormone secretion are still poorly understood. Here, the exocytotic machinery in pituitary cells is briefly reviewed in terms of the spatial organization of [Ca(2+)](i) elevation relative to the Ca(2+) sensor(s). PMID- 18406778 TI - Modeling plasma membrane and endoplasmic reticulum excitability in pituitary cells. AB - The response of gonadotrophs to secretagogues involves dose-dependent, complex dynamic patterns of electrical activity and inositol 1,4,5-trisphosphate (InsP(3))-induced Ca(2+) mobilization, including pulsatility and oscillations on multiple time scales from milliseconds to minutes. Detailed in vitro experiments have enabled the identification of key mechanisms that underlie the plasma membrane (PM) electrical excitability and endoplasmic reticulum (ER) calcium excitability. We summarize these findings and review computer simulations of a biophysical model that resynthesizes and couples these components and that reproduces quantitatively the observed time courses and dose-response characteristics, as well as effects of various pharamacological manipulations. The theory suggests that cytosolic calcium is the primary messenger in coordinating the PM and ER regenerative behaviors during ER depletion and refilling. PMID- 18406779 TI - Insulin-like Growth Factors as Regulators of Cell Motility Signaling Mechanisms. AB - Accumulating evidence indicates that the insulin-like growth factors (IGFs) function not only as mitogenic factors, but also as promoters of cell motility. In this article we review the current knowledge concerning the biochemical mechanisms whereby the IGFs activate cell motility. A key aspect of IGF stimulated cell motility is the ability of IGFs to promote actin polymerization at the leading edge of the cell. This effect of the IGFs is mediated by activation and autophosphorylation of the type I IGF receptor, followed by docking of insulin receptor substrate-1 (IRS-1), stimulation of phosphatidylinositol (PI) 3-kinase, and possibly activation of the small GTPase Rac. IGF-stimulated cell motility also requires the formation of new adhesions, a process associated with tyrosine phosphorylation of paxillin and focal adhesion kinase. Determining the biochemical mechanisms by which IGFs regulate cell motility should allow for a better understanding of bone remodeling, neurite outgrowth, tumor metastasis, placental formation, and skin and blood vessel repair. (c) 1997, Elsevier Science Inc. (Trends Endocrinol Metab 1997;8:1-6). PMID- 18406780 TI - Calcium regulation of adenylyl cyclase relevance for endocrine control. AB - A fundamental process in the hormonal regulation of body functions is the conversion of the intercellular signal into an intracellular signal. The first recognized intracellular messengers mediating the actions of hormones were calcium ions (Ca(2+)) and adenosine 3':5' monophosphate (cAMP), which is synthesized from ATP by adenylyl cyclase. Recent work on the structure of adenylyl cyclases has shown that these enzymes are individually tailored molecular machines controlled by diverse Ca(2+)-dependent mechanisms. These include allosteric regulation of enzyme activity through the Ca(2+)-receptor protein calmodulin, apparently direct actions of Ca(2+)on the cyclase catalytic moiety and phosphorylation/dephosphorylation by Ca(2+)-regulated protein kinases and protein phosphatases. This article is a brief review of the recent developments in the area of cyclase control that forecast a major revival of the interest in cAMP-Ca(2+)interactions. (c) 1997, Elsevier Science Inc. (Trends Endocrinol Metab 1997;8:7-14). PMID- 18406781 TI - Familial Congenital Hypothyroidism Caused by Abnormal and Bioinactive TSH due to Mutations in the beta-Subunit Gene. AB - Hereditary TSH deficiency is a rare autosomal recessive disease described in inbred Japanese families and in Greek and Brazilian kindreds. The TSH-beta subunit gene has been shown to be the site of mutations that will give rise to truncated proteins that cannot dimerize with the alpha subunit or, alternatively, will produce a mutated TSH that is present in the circulation of the affected patients, but it is biologically inactive. Characteristically, the patients with TSH-beta-subunit-defects are born with congenital hypothyroidism, with very low levels of serum thyroid hormones and serum thyroglobulin and, paradoxically, with serum TSH levels that are consistently undetectable or at very low levels. Goiter is not present at birth, but the low radioactive thyroid uptake will increase after bovine TSH stimulation. Other pituitary hormones responses to provocative tests are normal. The subunit levels are at high concentration and are significantly increased following TRH stimulation. In two kindreds, molecular biological studies have indicated mutations in two different sites of exon 2, generating a peptide that would not dimerize with subunits to synthesize TSH molecules. In one kindred, a truncated TSH-beta protein was translated that generated a biologically inactive but detectable serum TSH molecule. (c) 1997, Elsevier Science Inc. (Trends Endocrinol Metab 1997;8:15-20). PMID- 18406782 TI - Risk factors for thyroid cancer. AB - The potential risk factors for thyroid carcinoma development include genetic predisposition, exposure to therapeutic or environmental ionizing radiation, residence in areas of iodine deficiency or excess, history of preexisting benign thyroid disease, as well as hormonal and reproductive factors. In this review, we analyze some of the epidemiological data, as well as the possible molecular mechanisms by which certain environmental and genetic factors might predispose to thyroid tumorigenesis. (c) 1997, Elsevier Science Inc. (Trends Endocrinol Metab 1997; 8:20-25). PMID- 18406783 TI - Glucocorticoid feedback resistance. AB - Glucocorticoid feedback resistance can be inherited or locally acquired. The implications of these two forms of resistance for disease are strikingly different. The inherited form is characterized by enhanced adrenocortical function and hypercorticism to compensate for a generalized deficit in the glucocorticoid receptor gene, but these individuals lack symptoms of Cushing's syndrome. By contrast, resistance acquired at the level of the hypothalamic corticotropin-releasing hormone (CRH) neurons is linked to hypercorticism, which is not compensatory but overexposes the rest of the body and the brain to glucocorticoids. This cell-specific glucocorticoid resistance can be acquired by genetically predisposed individuals failing to cope with (early) life events and causes enhanced vulnerability to disease-specific actions of glucocorticoids. (c) 1997, Elsevier Science Inc. (Trends Endocrinol Metab 1997; 8:26-33). PMID- 18406784 TI - Roles of estrogen and progesterone in normal mammary gland development insights from progesterone receptor null mutant mice and in situ localization of receptor. AB - In contrast to most other organs, the development of normal mammary glands occurs mostly in the postnatal state and in a discontinuous fashion. In all species, the glands are composed of various cell types, and it is the epithelium, embedded in the fatty stroma (commonly known as the "fat pad"), that is targeted for proliferation and differentiation. Hormones, in particular the female sex steroids estrogen and progesterone, are absolutely essential for the proliferation of mammary epithelial cell. However, despite intensive efforts by many laboratories spanning nearly 4 decades, at present, the precise role of these steroids and their relative importance in mammary development remains unclear. This article reviews the general features and the effects of estrogen and progesterone on normal mammary gland development, followed by a discussion of our recent studies, which emphasize the primary importance of progesterone and progesterone receptor for the normal mammary epithelial cell proliferation and differentiation. (c) 1997, Elsevier Science Inc. (Trends Endocrinol Metab 1997;8:34-39). PMID- 18406785 TI - Thrombopoietin the primary regulator of platelet production. AB - Although the term thrombopoietin was first used nearly 40 years ago to describe the humoral regulator of platelet production, doubts surrounding its existence remained until the molecule was cloned 3 years ago. Using the recombinant protein, several investigators have shown that thrombopoietin influences all aspects of megakaryocyte development, from the hematopoietic stem cell to the mature platelet. The present review focuses on the discovery and characterization of this hormone, the initial stages of its clinical development, and some important yet unanswered questions of its molecular and cellular physiology. (Trends Endocrinol Metab 1997;8:45-50). (c) 1997, Elsevier Science Inc. PMID- 18406786 TI - Vanadium compounds biological actions and potential as pharmacological agents. AB - Vanadium is an element found in low concentrations in mammals, for which a function remains to be discovered. Over the past century, vanadium compounds have been suggested anecdotally as therapeutic agents for a variety of diseases. The discovery that vanadate inhibits various enzymes, in particular protein tyrosine phosphatases, and mimics many of the biological actions of insulin suggested a potential role in the therapy of diabetes mellitus. Successful use and an enhancement of insulin sensitivity in rodents and human diabetic subjects, as well as the finding that these agents are capable of stimulating metabolic effects while bypassing the insulin receptor and the early steps in insulin action, target these agents preferentially toward type II diabetes mellitus. Long term safety remains a major concern, as tissue accumulation and relative nonspecificity of enzyme inhibition may result in adverse effects. Continued research into mechanism of action, consequences of chronic administration, and improvement of specificity is warranted. Regardless of their ultimate success or failure as therapeutic agents, vanadium compounds continue to be useful probes of enzyme structure and function in various biological processes. (Trends Endocrinol Metab 1997;8:51-58). (c) 1997, Elsevier Science Inc. PMID- 18406787 TI - The autoimmune endocrinopathies the complexities continue to ravel. AB - The organ-specific autoimmune endocrinopathies constitute a group of disorders related to one another and to some other nonendocrine autoimmune organ-specific diseases. Although there are genetic links between these entities, there are genetic differences as well; the suggestion of autoantigenic crossreactivity has not borne fruit and does not appear to explain these associations. Each condition may be due to a defect(s) in specific antigen-presenting genes, with consequential effects on specific T lymphocyte activation. Genetic overlap may explain poly-endocrine autoimmune disease or the appearance of different maladies in other family members. The immune response is extremely complex, but the many elements involved, and molecules that can interdict them, provide some promise for potential new therapeutic immunomodulatory interventions. (Trends Endocrinol Metab 1997;8:59-63). (c) 1997, Elsevier Science Inc. PMID- 18406788 TI - The immunogenetics of autoimmune diabetes and autoimmune thyroid disease. AB - Although medical genetics is a well-developed area of interest, relatively little is known about the diseases caused by the combination of many genes. These multiinfluenced diseases include the autoimmune endocrine diseases. Recent advances in the techniques for whole-genome screening have shown a variety of loci that are linked to the development of insulin-dependent diabetes mellitus, and similar data are likely to be soon generated in autoimmune thyroid disease. Here, the authors survey the current state of genetic knowledge in these two areas and describe the investigative and analytical techniques that are now available. (Trends Endocrinol Metab 1997;8:63-70). (c) 1997, Elsevier Science Inc. PMID- 18406789 TI - Autoimmunity and type I diabetes. AB - Insulin-dependent diabetes mellitus (IDDM) is a T-cell-mediated autoimmune disease. The effector mechanisms essentially involve cytokine-mediated inflammation ultimately leading to beta-cell destruction. Several candidate autoantigens have been delineated for both the pathogenic T-cell response and the nonpathogenic antibody response used for disease prediction. Because of antigen spreading, it is not yet clear which of these antigens are involved in the triggering of the autoimmune response. In any case, this TH1 autoimmune response is amplified and perpetuated by an immune dysregulation involving TH2 cells. Both effector and regulatory mechanisms are placed under the tight control of major histocompatibility complex (MHC) and non-MHC genes. (Trends Endocrinol Metab 1997; 8:71-74). (c) 1997, Elsevier Science Inc. PMID- 18406790 TI - Autoimmune hypophysitis. AB - Autoimmune (lymphocytic) hypophysitis has emerged as a distinct and specific clinical and pathological disease entity. Although relatively rare compared with other autoimmune endocrine diseases, nearly a hundred cases have been described. The condition is much more common in females (9:1) and appears to have a particular predilection for the pregnant and postpartum states. The anterior pituitary, and less often the neurohypophysis, appear to be the target for inflammatory autoimmune destruction. During the evolution of the disease process, pituitary hyperfunction (usually hyperprolactinemia) has been noted. This disease should now be included in the differential diagnosis of pituitary disorders, especially in females presenting with pituitary enlargement, particularly if symptoms occur in temporal relationship to pregnancy. The disease may form part of the spectrum of the polyglandular autoimmune endocrine disorders. (Trends Endocrinol Metab 1997;8:74-80). (c) 1997, Elsevier Science Inc. PMID- 18406791 TI - Salmon calcitonin in the prevention and treatment of osteoporosis. AB - The principal calcitonins used therapeutically are porcine, human (synthetic), salmon (synthetic), and eel. Of these substances, salmon calcitonin is the most potent (Reginster 1991). Injectable forms of salmon calcitonin were first indicated for use in the treatment of Paget's disease in the United States and later received an indication for use in the treatment of postmenopausal osteoporosis. The injectable [intramuscular (IM) or subcutaneous (SC)] formulation of salmon calcitonin, however, has been associated with a high rate of noncompliance in elderly patients. The recently approved nasal-spray formulation has ameliorated the difficulties inherent in parenteral administration and is considered a suitable option as an alternative therapy in patients who are not appropriate candidates for estrogen replacement therapy. (Trends Endocrinol Metab 1997;8:89-92). (c) 1997, Elsevier Science Inc. PMID- 18406792 TI - Growth hormone treatment in children. AB - GH therapy increases final height in GH-deficient children. Short-term growth acceleration is also seen in children with many other causes of shortness. This review covers the diagnosis of GH-deficiency (GHD) and the details of GH treatment and its long-term results in GH-deficient patients and in those with other conditions, including "idiopathic short stature" and Turner syndrome. The efficacy of GH in enhancing adult stature in children with diagnoses other than GHD and Turner syndrome has not been established, and the only other indication for which it is approved by the U.S. Food and Drug Administration is chronic renal insufficiency. Broadening of the indications for GH use in childhood can only occur if supported by the results of carefully performed clinical trials. (Trends Endocrinol Metab 1997;8:92-97). (c) 1997, Elsevier Science Inc. PMID- 18406793 TI - Impotence in diabetes. AB - Impotence is more common in diabetic patients than in the normal population. This is due to the complications of diabetes, which damage the neurovascular bundle required for normal penile function. Minimally invasive methods of treatment have improved the outcome; however, the methods are still suboptimal. Recent advances in our understanding of the underlying mechanisms may lead to more non-invasive, targeted forms of treatment. (Trends Endocrinol Metab 1997;8:98-101). (c) 1997, Elsevier Science Inc. PMID- 18406794 TI - Insulin receptor knock-out mice. AB - Targeted mutagenesis of the insulin receptor gene in mice has yielded unexpected results. This article reviews recent findings and analyzes this animal model can further our understanding of the mechanism of insulin action and its impairment in non-insulin-dependent diabetes mellitus is analyzed. (Trends Endocrinol Metab 1997;8:101-104). Published 1997 by Elsevier Science Inc. PMID- 18406795 TI - In vivo Investigation of Hypothalamic Secretory Activity. AB - The finding that all pituitary hormones are released in a discrete pulsatile fashion and that the pulsatile properties of the pituitary hormone secretion are altered in some physiologic and pathologic conditions prompted the development of techniques designed to study the pattern of release and the regulation of secretion of the hypothalamic neuropeptides. This review describes the currently used techniques to assess hypothalamic hormone secretion. (Trends Endocrinol Metab 1997;8:105-111). (c) 1997, Elsevier Science Inc. PMID- 18406796 TI - Interactions between hormonal and environmental signals on hypothalamic neurons molecular mechanisms signaling environmental events. AB - It is axiomatic that the central nervous system must manage the integration of several environmental factors with steroid hormonal influences for the biologically adaptive performance of reproductive behavior. Launching from established behavioral investigations and from hormonal influences on gene function in the brain, we review here studies on how synaptic inputs and sex hormone influences codetermine hypothalamic gene expression. A particularly exciting implication of results on the ability of thyroid hormone receptors to interfere with estrogen receptor-dependent neuroendocrine function is that environmentally stimulated changes in thyroid hormone levels could influence hypothalamic transcriptional mechanisms important for behavior. If so, this would unite naturalistic environmental thinking with molecular neurobiological thinking important for the hypothalamic control of reproduction. (Trends Endocrinol Metab 1997;8:111-115). (c) 1997, Elsevier Science Inc. PMID- 18406797 TI - The logic of signaling from the cell surface to the nucleus. AB - The apparent imprecisions in the signal transduction mechanisms that couple specific hormone receptors to predictable nuclear transcription events have raised many issues for both biology and medicine. In this article, a fuzzy logic model is proposed to provide a means of formally describing signal transduction systems and making predictions about the effects of various combinations of inputs. The fuzzy logic model may provide opportunities mathematically to describe regulatory ensembles that are too complex to describe by conventional models. In addition, the nature of "fuzzy variables" provides insight into the paradoxes that underlie both imprecision and predictability in signal transduction. PMID- 18406798 TI - Receptor-mediated targeting of hormones to secretory granules: role of carboxypeptidase E. AB - Peptide hormones, neuropeptides, and other molecules such as the granins are specifically packaged into granules of the regulated secretory pathway and released in a calcium-dependent manner upon stimulation. Many of these molecules are synthesized as larger precursors (prohormones) that are processed to biologically active products within the granules. It has now become apparent that prohormones, proneuropeptides, and the granins contain conformation-dependent sorting signal motifs that facilitate their specific sorting and packaging into regulated secretory granules. Recently, a receptor to which these sorting signals bind has been identified as the membrane form of carboxypeptidase E (CPE) and localized to the Golgi apparatus, where sorting occurs, specifically at the trans Golgi network. In this article, we review the evidence for a sorting signal receptor-mediated mechanism for routing peptide hormones and prohormones to the regulated secretory granules. We also describe a mouse model, Cpe(fat), which has the CPE gene naturally mutated. Pituitary hormones were misrouted and secreted in an unregulated manner via the constitutive pathway in these Cpe(fat) mice, leading to endocrine disorders. PMID- 18406799 TI - Developmental and physiological effects of estrogen receptor gene disruption in mice. AB - Disruption of the estrogen receptor (ER) gene in mice causes infertility in both sexes. Infertility in female ER knockout (ERKO) mice results from altered development of accessory sex structures, disrupted endocrine physiology, and disrupted gametogenesis. Male accessory sex structures appear relatively normal, with infertility stemming from altered sexual behaviors and disrupted gametogenesis. These findings provide significant insights into the biological importance of the ER and suggest further areas for examining the impact of estrogens on reproductive biology. PMID- 18406800 TI - Homeodomain protein IDX-1: a master regulator of pancreas development and insulin gene expression. AB - The homeodomain protein IDX-1 appears to be a "master regulator" of pancreas development and beta-cell differentiation and function. In murine gene inactivation models and in a human subject with a homozygous mutation of the IDX 1 gene, the pancreas fails to develop. In the adult endocrine pancreas, IDX-1 is primarily expressed in beta cells, where it is a key factor in the upregulation of insulin gene transcription and appears to have a role in the regulation of the somatostatin, glucokinase, glucose transporter-2, and islet amyloid polypeptide genes. Recent studies also suggest a role for IDX-1 in the neogenesis and proliferation of beta cells. The observed functions of IDX-1 and its downregulation in parallel with insulin in glucose-toxicity models implicate IDX 1 as a potential factor contributing to the pathogenesis of diabetes mellitus. Future directions include the use of conditional gene inactivation to determine more precisely the role of IDX-1 throughout endocrine pancreas differentiation and the exploration of IDX-1 as a potential target for gene therapy of diabetes mellitus. PMID- 18406801 TI - Nuclear factor-kappa B repression in antiinflammation and immunosuppression by glucocorticoids. AB - The transcription factor nuclear factor-kappaB (NF-kappaB) directs transcription of a large number of key molecules in immunological and inflammatory responses. The recently discovered inhibition of transcriptional activity of this factor by the activated glucocorticoid receptor (GR) provides a molecular basis for the potent antiinflammatory and immunosuppressive properties of glucocorticoids. This repressive activity of the GR can function independently of transcriptional activity. Because it has been shown that certain steroid receptor ligands can differentially address transactivation and transrepression functions, it may be possible to develop ligands that specifically suppress NF-kappaB activity and, as a result, are more efficient in treatment of a variety of important chronic inflammatory diseases with less severe side effects than those of currently available drugs. PMID- 18406802 TI - Bone mass measurements: the case for selected screening? AB - Osteoporosis, a consequence of loss of bone mass, results in serious health and economic problems, particularly fractures of the hip and spine. Fractures are, in the majority of cases, the result of low bone mineral density (BMD). Technology is now available that allows accurate and sensitive measurement of BMD. The state of the art technology is dual x-ray absorptiometry (DXA), which, unfortunately, is not widely available and is relatively expensive. BMD measurement by DXA is the most individually objective predictor of fracture risk and, therefore, may be a reasonable method for universal screening of women. The pros and cons of mass screening to identify women with low bone mass and increased fracture risk are discussed. PMID- 18406803 TI - Spectrum of Adrenal Dysfunction in Patients with Acquired Immunodeficiency Syndrome Evaluation of Adrenal and Pituitary Reserve with ACTH and Corticotropin Releasing Hormone Testing. AB - Patients with acquired immunodeficiency syndrome (AIDS) have been reported to develop abnormalities of the endocrine system and in particular of the hypothalamic-pituitary-adrenal (HPA) axis. To define the abnormalities of HPA function in AIDS patients better, we performed ACTH and ovine corticotropin releasing hormone (oCRH) testing in a group of AIDS patients and oCRH testing in a group of healthy subjects. Our study found that in AIDS patients with normal ACTH testing, oCRH testing revealed a variety of subclinical abnormalities of ACTH and cortisol responses. Although we did not find frank adrenal insufficiency in any of these AIDS patients, it remains to be determined if any of the subclinical abnormalities we identified are predictive of clinically significant adrenal insufficiency; it may be that as AIDS patients live longer, the subclinical abnormalities will progress to adrenal insufficiency. (Trends Endocrinol Metab 1997;8:173-180). (c) 1997, Elsevier Science Inc. PMID- 18406804 TI - The role of Angiotensin-converting enzyme in blood pressure control, renal function, and male fertility. AB - Angiotensin-converting enzyme (ACE) is a zinc peptidase that plays a major role in the renin-angiotensin system. In mammals, the enzyme is present as two isozymes: a somatic form involved in blood-pressure regulation and a testis form of unknown function. Mice lacking ACE have been created and shown to have low systolic blood pressures and defects in renal development and function. These mice also have reduced male fertility, implicating the testis isozyme in reproductive function. (Trends Endocrinol Metab 1997;8:181-186). (c) 1997, Elsevier Science Inc. PMID- 18406805 TI - New concepts for treatment of non-insulin-dependent diabetes mellitus. AB - Non-insulin-dependent diabetes mellitus remains a major cause of morbidity and premature mortality in our community. Although potentially amenable to control by lifestyle modification, this is difficult to achieve in practice. Additional approaches using drugs that enhance insulin secretion, suppress hepatic glucose production, and increase insulin sensitivity are available, and new agents are being developed. The thiazolidinedione drugs hold particular promise as insulin sensitizing agents; however, at present, insulin administration is often also required. The importance of detection and treatment of risk factors for cardiovascular disease and the earlier detection and management of microvascular and infective complications remain of crucial importance. (Trends Endocrinol Metab 1997;8:187-191). (c) 1997, Elsevier Science Inc. PMID- 18406806 TI - Stress, acute hyperglycemia, and hyperlipidemia role of the autonomic nervous system and cytokines. AB - Stress is accompanied by metabolic alterations that could contribute to the etiology of diabetes mellitus, arteriosclerosis, and cardiovascular diseases; however, the mechanisms by which stress affects glucose and lipid metabolism remain to be resolved. Stress-induced effects on neurotransmission and interleukin-1 (IL-1) signaling rapidly produce hyperglycemia by increasing sympathetic outflow. Activation of the sympathetic nervous system can also rapidly stimulate lipolysis and hepatic triglyceride secretion. Furthermore, stress increases serum interleukin-6 (IL-6) and nerve growth factor (NGF) levels by activating neuroendocrine systems. IL-6 and NGF can rapidly increase lipolysis and hepatic triglyceride secretion without inducing hyperglycemia. The sympathetic nervous system does not mediate cytokine-induced hypertriglyceridemia. Thus, the central nervous system plays an important role in regulation of hepatic glucose and lipid metabolism via the sympathetic nervous system and cytokines. (Trends Endocrinol Metab 1997;8:192-197). (c) 1997, Elsevier Science Inc. PMID- 18406807 TI - Hormonal regulation of apoptosis an ovarian perspective. AB - Using the ovary as a model system for studying the hormonal regulation of apoptosis, recent studies have revealed that the survival of growing follicles is under the regulation of a complex array of hormones through endocrine, paracrine, autocrine, or juxtacrine mechanism in a development-dependent manner. More effort is needed, however, to identify tissue-specific factors required for the survival of ovarian somatic and germ cells at specific stage of development. New insights based on characterization of conserved apoptotic effectors, both extracellular and intracellular, have suggested that apoptosis in ovarian cells may be mediated by apoptotic programs common to other cells but using specific members of the death domain proteins as well as ced-9/Bcl-2 and ced-3/ICE caspase families of genes. Future studies may provide new therapeutic modalities for different ovarian diseases caused by aberrant regulation of apoptosis in ovarian cells, including premature ovarian failure and polycystic ovarian syndrome. (Trends Endocrinol Metab 1997;8:207-213). (c) 1997, Elsevier Science Inc. PMID- 18406808 TI - Anabolic effects of parathyroid hormone on bone. AB - Parathyroid hormone (PTH) is classically considered to be a bone catabolic agent; however, when delivered intermittently at low doses, PTH potently stimulates cortical and/or trabecular bone growth in animal and clinical studies. Understanding the mechanism of PTH's osteopenic actions has led to the making of small, novel, and possibly noninjectable PTH analogues that can build biomechanically normal bone in osteopenic rats and are promising candidates for treating osteoporosis in humans. PMID- 18406809 TI - Genes and osteoporosis. AB - Osteoporosis is a common disease associated with reduced bone mineral density, affecting up to 40% of women and 12% of men at some point during life. Although osteoporosis is multifactorial, genetic factors play an important role in the pathogenesis of osteoporosis, as up to 75% of the variance in bone mass-a major risk factor for osteoporotic fracture-is genetically determined. Segregation analysis of bone mass in families has suggested a model whereby bone mass is under the control of a large number of genes with small effect, rather than a few genes with large effect. Although the molecular-genetic basis by which bone mass is regulated is incompletely defined, polymorphisms of several candidate genes have been linked to bone mass in clinical studies. Polymorphisms in the gene encoding the vitamin D receptor have been extensively investigated with differing results, and interleukin-6, transforming growth factor beta, and the estrogen receptor have been associated with bone mass in isolated studies. Recent work has identified a polymorphism in the binding site for the transcription factor Sp1 in the collagen type I alpha 1 gene. This polymorphism is associated with bone mass and osteoporotic fracture in three distinct populations, suggesting that the polymorphism may act as a marker for low bone density and fracture risk. Further studies will uncover many new genes and candidate loci that relate to bone mass. This information may be of value in defining new therapeutic targets in the prevention and management of osteoporosis and in developing genetic tests to assess osteoporotic fracture risk. PMID- 18406810 TI - The economics of osteoporosis prevention. AB - Until recently, it was generally assumed that the prevention of a significant proportion of osteoporotic fractures could only be achieved through the use of hormone replacement therapy soon after menopause. It is likely to be more cost effective, however, if antifracture therapy could be undertaken among older, higher risk populations. Recent evidence suggests that there are a number of effective interventions that can be used among the over-65 age group, and the economic evidence supports the view that this is likely to be the best way forward for fracture prevention. PMID- 18406811 TI - Bone and mineral metabolism in African Americans. AB - Important differences exist in the metabolism of bone and mineral and the vitamin D endocrine system between whites and African Americans and include rate o f skeletal remodeling, bone mass, and vitamin D metabolism. A higher bone mineral density (BMD) in African Americans is associated with a diminished incidence o f osteoporosis and fractures. Serum 17beta-estradiol and the rate of GH secretion are higher in black than in white men, but there is no racial difference in women in this regard. The mechanisms for reduced rate o f skeletal remodeling and for greater BMD in blacks are not known, but diminished rate of skeletal remodeling could be a contributing factor for greater bone mass. Reduction in serum 25 hydroxyvitamin D in blacks is attributed to increased skin pigment and to diminished dermal production of vitamin D(3) and consequent decreased hepatic synthesis o f the metabolite. There is no evidence that alteration of the vitamin D endocrine system contributes to or is responsible for racial differences in skeletal remodeling and bone mass. Black infants, however, are at risk for developing vitamin D-deficient rickets, particularly when breast-fed. PMID- 18406812 TI - Vitamin D and hematopoiesis. AB - Analysis of the nonclassic actions of vitamin D(3) has highlighted a wide range of target tissues for the hormone 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. Systemic or locally produced 1,25(OH)(2)D(3) may play a role in modulating cell development processes such as hematopoiesis. The mechanisms by which 1,25(OH)(2)D(3) achieves this are discussed in this review. In particular, data from our laboratories suggest that 1,25(OH)(2)D(3) does not provide a deterministic signal for monocyte differentiation. Rather, the hormone acts as a permissive agent for myeloid precursor cells to enter a genetically determined terminal maturation pathway. The effiacy of 1,25(OH)(2)D(3) in leukemia therapy has been improved by the development of novel vitamin D analogues that have potent antiproliferative activity and low hypercalcemic side effects. Another solution to the problem of side effects is to enhance specifically the antiproliferative effects of 1,25(OH)(2)D(3). A novel mechanism within hematopoiesic cells that governs their responsiveness to the antiproliferative/differentiative actions of 1,25(OH)(2)D(3) outlined. PMID- 18406813 TI - Angiotensin II signal transduction: Stimulation of multiple mitogen-activated protein kinase pathways. AB - Angiotensin II (AngII)( *) is the main effector of the renin-angiotensin system and influences cell growth, migration, differentiation, and apoptosis. The mitogen-activated protein (MAP) kinase family of serine-threonine kinases integrates diverse extracellular stimuli and is involved in the regulation of cell growth and apoptosis. AngII regulates activity of the MAP kinase family members extracellular regulated kinases (ERK1/2) and c-Jun N-terminal kinase (JNK). This review discusses the signal transduction pathways leading to ERK1/2 and JNK activation by AngII. PMID- 18406814 TI - Placing progesterone in the apoptotic pathway. AB - Progesterone regulates the viability of cells of several different reproductive tissues, including the uterus, breast, corpus luteum, and ovarian follicle (that is, granulosa cells). Progesterone's antiapoptotic actions are thought to be mediated through the progesterone receptor because the progesterone antagonist RU 486 attenuates progesterone's action. Uterine, mammary, and luteal cells express the classic nuclear progesterone receptor, whereas granulosa cells of developing and mature follicles do not. This review presents data that suggest that progesterone maintains granulosa cell viability through a nongenomic mechanism. PMID- 18406815 TI - Yeast molecular chaperones and the mechanism of steroid hormone action. AB - The yeast Saccharomyces cerevisiae has become a valuable tool for the analysis of steroid action in vivo. Results from studies using yeast have been productive for understanding the role of molecular chaperones, especially Hsp90 and its cochaperones, in regulating the activation of heterologously expressed steroid hormone receptors. The methodology for these investigations involves assaying receptor function in yeast strains with deletions or mutations in the genes encoding molecular chaperone proteins. The information obtained thus far indicates that the Hsp90 chaperone machine functions in vivo to maintain steroid receptors in a high-affinity hormone-binding conformation and also in downstream events of the activation pathway. PMID- 18406816 TI - How obesity causes diabetes in Zucker diabetic fatty rats. AB - The mechanisms of adipogenic diabetes in Zucker diabetic fatty (ZDF) rats, a model of obesity complicated by diabetes, are reviewed. In ZDF rats, a mutation in the leptin receptor, OB-R, is associated with leptin resistance, obesity, and increased fat content of islets. Exaggerated nitric oxide (NO) generation, attributed to high intracellular levels of long-chain fatty acids, impairs beta cell function and prevents their compensation for obesity-induced diabetes. The resulting diabetic hyperglycemia can be completely prevented by agents that inhibit NO production. PMID- 18406817 TI - Thyroid function testing in psychiatric illness: Usefulness and limitations. AB - The assessment of thyroid function in psychiatric patients may be obscured by several effects of the psychiatric condition on both thyroid hormone and TSH levels. Acute psychiatric decompensation may result in elevation in total T(4) and free T(4) index, and less frequently in hypothyroxinemia. In addition, psychiatric illnesses can cause suppressed TSH levels, blunted TSH response to thyrotropin-releasing hormone (TRH) (particularly in depression), and elevated TSH values that may result in diagnostic errors. Even though mechanisms similar to the ones responsible for thyroid function test changes in other nonthyroidal illness could account for some of these abnormalities, other mechanisms involving dysregulation of hypothalamic-pituitary function seem to play an important role. TRH stimulation testing has also been used for the diagnosis and prognosis of some psychiatric disorders. This test, however, appears to have low sensitivity and specificity and little clinical usefulness for this purpose and may be replaced by basal TSH levels determined by highly sensitive assays. In this review, in addition to discussing the usefulness and limitations of thyroid function tests in the setting of a psychiatric condition, we provide a stepwise approach, using sensitive TSH as a first-line test in the assessment of thyroid function in psychiatric patients. PMID- 18406818 TI - Insulinlike growth factor-binding protein proteolysis an emerging paradigm in insulinlike growth factor physiology. AB - In biologic fluids, insulinlike growth factors (IGF-I and IGF-II) are bound to high-affinity insulinlike growth factor binding proteins (IGFBPs) of which seven have now been identified (IGFBPs 1-7). In a variety of biologic fluids, several IGFBPs undergo proteolytic degradation. Such degradation can lead to increased IGF bioavailability at the cell surface, facilitating receptor interactions. Herein, recent data identifying several IGFBP-degrading proteinases and their effects on IGF bioactivity is reviewed, and how IGFBP proteolysis is regulated by IGFs and IGFBPs, as well as how IGFBP cleavage analysis provides insights into the structure and function of IGFBPs, is explored. (Trends Endocrinol Metab 1997;8:299-306). (c) 1997, Elsevier Science Inc. PMID- 18406819 TI - Insulin receptor what role in breast cancer? AB - It is commonly believed that the insulin receptor mainly mediates the metabolic effects of insulin, whereas the closely related IGF-I receptor is considered a major factor for the regulation of cell proliferation. Experimental and epidemiological evidence indicates, however, that insulin and insulin receptors may play an important role in breast cancer. This article reviews evidence indicating that (a) insulin receptors are overexpressed in human breast cancer, (b) insulin stimulates growth in breast cancer cells, (c) cells transfected with human insulin receptor may acquire a ligand-dependent transformed phenotype, and (d) breast cancer is associated with insulin resistance and hyperinsulinemia. These findings may open new possibilities in breast cancer prevention, prognosis assessment, and therapy. (Trends Endocrinol Metab 1997; 8:306-312). (c) 1997, Elsevier Science Inc. PMID- 18406820 TI - Estrogen receptors and cell proliferation in breast cancer. AB - Most of the actions of estrogens on the normal and abnormal mammary cells are mediated via estrogen receptors (ERs), including control of cell proliferation; however, there are also alternative pathways of estrogen action not involving ERs. Estrogens control several genes and proteins that induce the cells to enter the cell cycle (protooncogenes, growth factors); estrogens also act on proteins directly involved in the control of the cell cycle (cyclins), and moreover, estrogens stimulate the response of negative cell cycle regulators (p53, BRCA1). The next challenge for researchers is elucidating the integration of the interrelationships of the complex pathways involved in the control of cell proliferation. This brief review focuses on the mechanisms of estrogen action to control cell proliferation and the clinical implications in breast cancer. (Trends Endocrinol Metab 1997;8:313-321). (c) 1997, Elsevier Science Inc. PMID- 18406821 TI - Insect neuropeptides and their receptors new leads for medical and agricultural applications. AB - Diversification of messenger and receptor molecules is the result of evolution; however, the principles of intercellular signaling mechanisms are very similar in all metazoans. Recent discoveries of insect peptides provide new leads for applications in medicine and agriculture. (Trends Endocrinol Metab 1997;8:321 326). (c) 1997, Elsevier Science Inc. PMID- 18406822 TI - Protein kinase C and mammalian spermatozoa acrosome reaction. AB - The presence of protein kinase C (PKC) in mammalian sperm was demonstrated by enzymatic activity assay and immunohistochemistry at the light and electron microscopy levels. The sperm head PKC is localized in the acrosome, equatorial segment, and postacrosomal region. In the flagellum, PKC is associated with the segmented column of the neck and is distributed along the mid, principal, and end pieces. Immunoreactive sites are observed in patches along the axoneme and outer dense fibers and are evenly distributed between these regions. Functional studies suggest the involvement of PKC in flagellar motility and acrosome reaction. The cross-talk between the signaling cascades that operate during sperm activation is discussed. PMID- 18406823 TI - Sex-determining genes. AB - Recently, a number of genes have been identified that are associated with a failure of human sex determination, including WT1, DAX-1, SOX9, ATRX, and the Y linked testis determination gene, SRY. Most cases of human sex reversal, XY females and XX males, do not, however, appear to be caused by mutations in these genes. This review highlights recent advances in this field and discusses the prospects of identifying genes in the sex-determining pathway. PMID- 18406824 TI - Aldosterone secretion: a molecular perspective. AB - The major mineralocorticoid hormone aldosterone is secreted from the zona glomerulosa of the adrenal cortex. Aldosterone is synthesized from cholesterol via a series of hydroxylations and oxidations. The enzymes involved in these reactions are mostly members of the cytochrome P450 superfamily. The final steps of this pathway, the conversion of 11-deoxycorticosterone (DOC) to aldosterone, require conversion via the intermediates 18-hydroxy-DOC or corticosterone and 18 hydroxycorticosterone. There are significant differences between species in the number of genes that encode the P450(11beta)-related enzymes (CYP11B) involved in these steps and the zonal distribution of their expression. One enzyme is capable of 11-hydroxylation, 18-hydroxylation, and 18-oxidation of DOC to aldosterone. The genetic basis of four diseases-congenital adrenal hyperplasia due to 11beta hydroxylase deficiency, glucocorticoid-remediable aldosteronism, aldosterone synthase deficiency type I and type II-is explicable by mutations in these cytochrome P450(11beta)-related genes. PMID- 18406825 TI - Growth-inhibitory actions of analogues of Luteinizing Hormone Releasing Hormone on tumor cells. AB - The expression of LHRH and its receptor has been demonstrated in a number of human malignant tumors, including cancers of the breast, ovary, endometrium, and prostate. These findings suggest the presence of an autocrine regulatory system based on LHRH. Dose-dependent antiproliferative effects of LHRH agonists in cell lines derived from these cancers have been observed by various investigators. LHRH antagonists also have marked antiproliferative activity in most of the ovarian, breast, and endometrial cancer cell lines tested, indicating that the dichotomy of LHRH agonists and antagonists might not apply to the LHRH system in cancer cells. Findings from our laboratories suggest that the classical LHRH receptor signal-transduction mechanisms, known to operate in the pituitary, are not involved in the mediation of antiproliferative effects of LHRH analogues in cancer cells. Results obtained by several groups, including ours, instead suggest that LHRH analogues interfere with the mitogenic signal transduction of growth factor receptors and related oncogene products associated with tyrosine kinase activity. The pharmacological exploitation of these direct antiproliferative actions of LHRH analogues might provide new therapeutic approaches to these cancers. PMID- 18406826 TI - Genetic basis of familial neurohypophyseal diabetes insipidus. AB - Familial neurohypophyseal diabetes insipidus (FNDI) is an autosomal dominant (adFNDI) or X-linked recessive (xrFNDI) disorder characterized by the development in early childhood of an irreversible deficiency of arginine vasopressin (AVP) secretion. Autopsy data in adFNDI reveal selective destruction of the posterior pituitary magnocellular neurons that normally produce the hormone. These abnormalities are due to a variety of mutations in the gene that encodes the AVP neurophysin II precursor. Each one predicts a change in the primary structure of the preprohormone, and all but one are of a type known or reasonably presumed to impair the folding and cellular trafficking of the preprohormone. This pattern and the uniform clinical characteristics of adFNDI suggest that the disease is due to the production of a mutant precursor that is toxic for magnocellular neurons, because it cannot be folded, processed, or otherwise disposed of efficiently. Although the gene responsible for xrFNDI has not yet been cloned, the striking clinical similarities between adFNDI and xrFNDI suggest that similar pathophysiologic mechanisms may be involved. PMID- 18406827 TI - Chromatin modulation of glucocorticoid and progesterone receptor activity. AB - Steroid hormone receptors are ligand-activated transcription factors that enhance or repress gene expression. They act by binding to target sites within the promoters of genes assembled as chromatin. Chromatin structure is modified in response to steroid hormones and represents a critical step in steroid receptor signaling. Recent experiments demonstrate that the progesterone and glucocorticoid receptors are differentially influenced by this arrangement of DNA and histones. One of the most important developments in the steroid hormone receptor field has been the identification of coactivators and cointegrators, some of which are histone acetyltransferases. These proteins appear to play an important role in mediating ligand activation of transcription, although their exact role on chromatin templates is undefined. PMID- 18406828 TI - gp130-Related cytokines and their receptors in the pituitary. AB - gp130-Related cytokines such as interleukin-6 and leukemia-inhibitory factor (LIF) act on the adenohypophysis in a paracrine manner, affecting both its differentiation and the function of specific cell types, notably the proopiomelanocortin (POMC) cells. They act on POMC cells in synergism with corticotrophin-releasing hormone, inducing ACTH secretion. gp130-Related cytokines as well as their receptors are expressed in the pituitary. LIF knockout mice show reduced stress-induced ACTH secretion, which can be restored by LIF replacement, suggesting a physiologic role for LIF. PMID- 18406829 TI - Somatostatin receptors. AB - The diverse biological effects of somatostatin (SRIF) are mediated by a family of G protein-coupled receptors (termed sst) that are encoded by five nonallelic genes located on separate chromosomes. The receptors can be further divided into two subfamilies: sst(2,3,5) react with octapeptide and hexapeptide SRIF analogues and belong to one subclass; sst(1,4) react poorly with these compounds and fall into another subclass. This review focuses on the molecular pharmacology and function of these receptors, with particular emphasis on the ligand-binding domain, subtype-selective analogues, agonist-dependent receptor regulation and desensitization responses, subtype-specific effector coupling, and signal transduction pathways responsible for inhibiting cell secretion and cell growth or induction of apoptosis. PMID- 18406830 TI - Alternative splicing of mRNA as a mode of endocrine regulation. AB - Alternative splicing of messenger RNA (mRNA) is a means of regulating gene expression and occurs in many genes of the endocrine system. This review covers an introduction into mRNA splicing and the mechanisms and regulation of alternative splicing. Some examples are discussed in which alternatively spliced genes encode functionally distinct proteins. Evidence that hormones and other metabolic signals may regulate alternative splicing events is reviewed, and potential mechanisms are considered. PMID- 18406831 TI - Disposable biosensor and biocatalysis of horseradish peroxidase based on sodium alginate film and room temperature ionic liquid. AB - Composite material based on room temperature ionic liquid (RTIL) N butylpyridinium hexafluorophosphate (BPPF(6)), sodium alginate (SA), and graphite was used to construct a novel horseradish peroxidase (HRP) biosensor for the determination of H(2)O(2). The morphology of the as-prepared biosensor was characterized by scanning electron microscopy (SEM). Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to characterize the process of the performance of the biosensor. Parameters affecting the performance of the biosensor, including the concentrations of o-aminophenol (OAP) and HRP and the pH value of substrate solution, were optimized. Under the optimized experimental conditions, H(2)O(2) could be detected in a linear calibration range of 1.0 to 6.0 microM with a correlation coefficient of 0.9847 (n=7) and a detection limit of 0.5 microM at a signal/noise ratio of 3. The prepared biosensor not only had economic and disposable property but also showed good detection precision, bioactivity, storage stability, and reproducibility. PMID- 18406832 TI - Coupling of fully automated chip-based electrospray ionization to high-capacity ion trap mass spectrometer for ganglioside analysis. AB - NanoMate robot was coupled to a high-capacity ion trap (HCT) mass spectrometer to create a system merging automatic chip-based electrospray ionization (ESI) infusion, ultrafast ion detection, and multistage sequencing at superior sensitivity. The interface between the NanoMate and HCT mass spectrometer consists of an in-laboratory constructed mounting device that allows adjustment of the robot position with respect to the mass spectrometer inlet. The coupling was optimized for ganglioside (GG) high-throughput analysis in the negative ion mode and was implemented in clinical glycolipidomics for identification and structural characterization of anencephaly-associated species. By NanoMate HCT mass spectrometry (MS), data corroborating significant differences in GG expression in anencephalic versus age-matched normal brain tissue were collected. The feasibility of chip-based nanoESI HCT multistage collision-induced dissociation (CID MS(n)) for polysialylated GG fragmentation and isomer discrimination was tested on a GT1 (d18:1/18:0) anencephaly-associated structure. MS(2)-MS(4) obtained by accumulating scans at variable fragmentation amplitudes gave rise to the first fragmentation patterns from which the presence of GT1b structural isomer could be determined unequivocally without the need for supplementary investigation by any other analytical or biochemical methods. PMID- 18406833 TI - Luteal supplementation in in vitro fertilization: more questions than answers. AB - OBJECTIVE: To update clinicians on different regimens of luteal phase supplementation in IVF-stimulated cycles and to identify areas that need further research in this subject. DESIGN: Literature review and critical analysis of published studies on luteal phase supplementation during the last 20 years. CONCLUSION(S): Luteal phase supplementation in IVF-stimulated cycles, both in gonadotropin releasing hormone agonist and antagonist protocols, is considered an essential requirement for optimal success rates. The date of initiation and discontinuation of supplemented hormones is not adequately studied in the literature. In most major controlled and randomized studies, there are no significant differences in success rates with progesterone supplementation alone, progesterone and estradiol, progesterone and human chorionic gonadotropin, and human chorionic gonadotropin alone. Success rates seem similar with intramuscular and vaginal progesterone administration with patient preference for the vaginal route. The optimal dose of progesterone has not been studied in a scientific way in the literature. The use of gonadotropin releasing hormone agonists for luteal phase supplementation in antagonist cycles appears to be promising, and is worthy of further investigation. PMID- 18406834 TI - Asherman syndrome--one century later. AB - OBJECTIVE: To provide an update on the current knowledge of Asherman syndrome. DESIGN: Literature review. SETTING: The worldwide reports of this disease. PATIENT(S): Patients with Asherman syndrome who presented with amenorrhea or hypomenorrhea, infertility, or recurrent pregnancy loss. INTERVENTION(S): Hysteroscopy and hysteroscopic surgery have been the gold standard of diagnosis and treatment respectively for this condition. MAIN OUTCOME MEASURE(S): The etiology, pathology, symptomatology, diagnosis, treatment, and reproductive outcomes were analyzed. RESULT(S): This syndrome occurs mainly as a result of trauma to the gravid uterine cavity, which leads to the formation of intrauterine and/or intracervical adhesions. Despite the advances in hysteroscopic surgery, the treatment of moderate to severe Asherman syndrome still presents a challenge. Furthermore, pregnancy after treatment remains high risk with complications including spontaneous abortion, preterm delivery, intrauterine growth restriction, placenta accrete or praevia, or even uterine rupture. CONCLUSION(S): The management of moderate to severe disease still poses a challenge, and the prognosis of severe disease remains poor. Close antenatal surveillance and monitoring are necessary for women who conceive after treatment. PMID- 18406835 TI - Progesterone supplementation during the luteal phase and in early pregnancy in the treatment of infertility: an educational bulletin. AB - Exogenous P supplementation is a common element of treatment regimens for infertility, particularly those relating to the assisted reproductive technologies. PMID- 18406836 TI - Guidelines for development of an emergency plan for in vitro fertilization programs. PMID- 18406837 TI - Opinions of infertile Turkish women on gamete donation and gestational surrogacy. AB - OBJECTIVE: To determine the approval levels of infertile Turkish women concerning gamete donation and gestational surrogacy. DESIGN: Opinion survey. SETTING: Assisted reproductive treatment center at Gulhane Military Medical Academy. PATIENT(S): 368 women who had applied for infertility treatment. INTERVENTION(S): The patients were asked to answer a questionnaire that included questions about the patient's sociodemographic status, previous medical history with infertility treatment, and opinions on gamete donation and gestational surrogacy. MAIN OUTCOME MEASURE(S): Opinions of patients concerning gamete donation and gestational surrogacy. RESULT(S): Our data in this first study on infertile Turkish women show that some patients approve of gamete donation (23.3% for accepting oocytes and 3.4% for accepting sperm) and gestational surrogacy (15.1%). CONCLUSION(S): Donation and surrogacy are alternate treatments for the serious condition called infertility. As there are patients who would like to use these treatments, each of these patients should have the right to try any of them. PMID- 18406838 TI - South Africa in the spotlight. PMID- 18406839 TI - False logic in new UK screening plans. PMID- 18406840 TI - International action needed on dengue. PMID- 18406841 TI - Countdown to 2015: a report card on maternal, newborn, and child survival. PMID- 18406842 TI - Making the Countdown count. PMID- 18406843 TI - Parliamentarians: leading the change for maternal, newborn, and child survival? PMID- 18406844 TI - Delivery of MDG 5 by active management with data. PMID- 18406845 TI - Sexual and reproductive health: completing the continuum. PMID- 18406846 TI - The village-based midwife programme in Indonesia. PMID- 18406847 TI - Getting political: fighting for global health. PMID- 18406848 TI - Delivering for women and children. PMID- 18406851 TI - Group Carso, health philanthropy, and tobacco. PMID- 18406852 TI - Group Carso, health philanthropy, and tobacco. PMID- 18406853 TI - Cardiotoxicity associated with sunitinib. PMID- 18406854 TI - Cardiotoxicity associated with sunitinib. PMID- 18406856 TI - When medicine meets politics. PMID- 18406857 TI - The UK's health system: gnu needed. PMID- 18406858 TI - Academy of Medical Sciences approaches its troublesome teens. PMID- 18406859 TI - Countdown to 2015 for maternal, newborn, and child survival: the 2008 report on tracking coverage of interventions. AB - BACKGROUND: The Countdown to 2015 for Maternal, Newborn, and Child Survival initiative monitors coverage of priority interventions to achieve the Millennium Development Goals (MDG) for reduction of maternal and child mortality. We aimed to report on 68 countries which have 97% of maternal and child deaths worldwide, and on 22 interventions that have been proven to improve maternal, newborn, and child survival. METHODS: We selected countries with high rates of maternal and child deaths, and interventions with the most potential to avert such deaths. We analysed country-specific data for maternal and child mortality and coverage of selected interventions. We also tracked cause-of-death profiles; indicators of nutritional status; the presence of supportive policies; financial flows to maternal, newborn, and child health; and equity in coverage of interventions. FINDINGS: Of the 68 priority countries, 16 were on track to meet MDG 4. Of these, seven had been on track in 2005 when the Countdown initiative was launched, three (including China) moved into the on-track category in 2008, and six were included in the Countdown process for the first time in 2008. Trends in maternal mortality that would indicate progress towards MDG 5 were not available, but in most (56 of 68) countries, maternal mortality was high or very high. Coverage of different interventions varied widely both between and within countries. Interventions that can be routinely scheduled, such as immunisation and antenatal care, had much higher coverage than those that rely on functional health systems and 24-hour availability of clinical services, such as skilled or emergency care at birth and care of ill newborn babies and children. Data for postnatal care were either unavailable or showed poor coverage in almost all 68 countries. The most rapid increases in coverage were seen for immunisation, which also received significant investment during this period. INTERPRETATION: Rapid progress is possible, but much more can and must be done. Focused efforts will be needed to improve coverage, especially for priorities such as contraceptive services, care in childbirth, postnatal care, and clinical case management of illnesses in newborn babies and children. PMID- 18406860 TI - Mind the gap: equity and trends in coverage of maternal, newborn, and child health services in 54 Countdown countries. AB - BACKGROUND: Increasing the coverage of key maternal, newborn, and child health interventions is essential if Millennium Development Goals (MDG) 4 and 5 are to be reached. We have assessed equity and trends in coverage rates of a key set of interventions through a summary index, to provide overall insight into past performance and progress perspectives. METHODS: Data from household surveys from 54 countries in the Countdown to 2015 for Maternal, Newborn and Child Survival initiative during 1990-2006 were used to compute an aggregate coverage index based on four intervention areas: family planning, maternal and newborn care, immunisation, and treatment of sick children. The four areas were given equal weight in the computation of the index. Standard measures were applied to assess current levels and trends in the coverage gap measure by wealth quintile. FINDINGS: The overall size of the coverage gap ranged from less than 20% in Tajikistan and Peru to over 70% in Ethiopia and Chad, with a mean of 43% for the most recent surveys in the 54 countries. Large intracountry differences were noted, with a country mean coverage gap of 54% for the poorest quintiles of the population and 29% for the wealthiest. Differences between the poorest and the wealthiest were largest for the maternal and newborn health intervention area and smallest for immunisation. In 40 countries with more than one survey, the coverage gap had decreased by an average of 0.9 percentage points per year since the early 1990s. Declines greater than 2 percentage points per year were seen in only three countries after 1995: Cambodia, Mozambique, and Nepal. Country inequity patterns were remarkably persistant over time, with only gradual changes from top inequity (disproportionately smaller gap for the wealthiest) in countries with coverage gaps exceeding 40%, to linear patterns and bottom inequity (disproportionately greater gap for the poorest) in surveys with gaps below 40%. INTERPRETATION: Despite most Countdown countries having made gradual progress since 1990, coverage gaps for key interventions remain wide and, in most such countries, the pace of decline needs to be more than doubled to reach levels of coverage of these and other interventions needed in the context of MDG 4 and 5. In general, in-country patterns of inequality are consistant and change only gradually if at all, which has implications for the targeting of interventions. PMID- 18406861 TI - Countdown to 2015: assessment of donor assistance to maternal, newborn, and child health between 2003 and 2006. AB - BACKGROUND: To track donor assistance to maternal, newborn, and child health related activities is necessary to assess progress towards Millennium Development Goals 4 and 5 and to foster donor accountability. Our aim was to analyse aid flows to maternal, newborn, and child health for 2005 and 2006 and trends between 2003 and 2006. METHODS: We analysed and coded the complete aid activities database for 2005 and 2006 with methods that we developed previously to track official development assistance. For the 68 Countdown priority countries, we report two indicators for use in monitoring donor disbursements: official development assistance to child health per child and official development assistance to maternal and neonatal health per livebirth. FINDINGS: Donor disbursements increased from US$2119 million in 2003 to $3482 million in 2006; funding for child health increased by 63% and that for maternal and newborn health increased by 66%. In the 68 priority countries, child-related disbursements increased from a mean of $4 per child in 2003 to $7 per child in 2006; disbursements for maternal and neonatal health increased from $7 per livebirth in 2003 to $12 per livebirth in 2006. Nonetheless, disbursements fell in some countries. After adjustment for other determinants, countries with higher under-5 mortality received more official development assistance per child, but official development assistance to maternal and newborn health did not seem to be well targeted towards countries with the greatest maternal health needs. INTERPRETATION: Donor resource tracking should be continued to help hold donors accountable and encourage targeting of resources to countries with greatest needs. PMID- 18406862 TI - Child survival gains in Tanzania: analysis of data from demographic and health surveys. AB - BACKGROUND: A recent national survey in Tanzania reported that mortality in children younger than 5 years dropped by 24% over the 5 years between 2000 and 2004. We aimed to investigate yearly changes to identify what might have contributed to this reduction and to investigate the prospects for meeting the Millennium Development Goal for child survival (MDG 4). METHODS: We analysed data from the four demographic and health surveys done in Tanzania since 1990 to generate estimates of mortality in children younger than 5 years for every 1-year period before each survey back to 1990. We estimated trends in mortality between 1990 and 2004 by fitting Lowess regression, and forecasted trends in mortality in 2005 to 2015. We aimed to investigate contextual factors, whether part of Tanzania's health system or not, that could have affected child mortality. FINDINGS: Disaggregated estimates of mortality showed a sharp acceleration in the reduction in mortality in children younger than 5 years in Tanzania between 2000 and 2004. In 1990, the point estimate of mortality was 141.5 (95% CI 141.5-141.5) deaths per 1000 livebirths. This was reduced by 40%, to reach a point estimate of 83.2 (95% CI 70.1-96.3) deaths per 1000 livebirths in 2004. The change in absolute risk was 58.4 (95% CI 32.7-83.8; p<0.0001). Between 1999 and 2004 we noted important improvements in Tanzania's health system, including doubled public expenditure on health; decentralisation and sector-wide basket funding; and increased coverage of key child-survival interventions, such as integrated management of childhood illness, insecticide-treated nets, vitamin A supplementation, immunisation, and exclusive breastfeeding. Other determinants of child survival that are not related to the health system did not change between 1999 and 2004, except for a slow increase in the HIV/AIDS burden. INTERPRETATION: Tanzania could attain MDG 4 if this trend of improved child survival were to be sustained. Investment in health systems and scaling up interventions can produce rapid gains in child survival. PMID- 18406863 TI - Assessment of the health system and policy environment as a critical complement to tracking intervention coverage for maternal, newborn, and child health. AB - In 2008, the Countdown to 2015 initiative identified 68 priority countries for action on maternal, newborn, and child health. Much attention was paid to monitoring country-level progress in achieving high and equitable coverage with interventions effective in reducing mortality of mothers, newborn infants, and children up to 5 years of age. To have a broader understanding of the environment in which health services are delivered and health outcomes are produced is essential to increase intervention coverage. Programmes to address MNCH rely on health systems to generate information needed for effective decisions and to achieve the expected outcomes. Governance and leadership are needed throughout the process not only to create policies and implement them but also to assure quality and efficiency of care, to finance health services sufficiently and in an equitable way, and to manage the health workforce. We present a systematic approach to assess the wider health system and policy environment needed to achieve positive outcomes for maternal, newborn, and child health. We report on results from 13 indicators and show gaps in policy adoption as well as weaknesses in other health system building blocks. We identify areas for future action in measurement of key indicators and their use to support decision making. We hope that this information will provide an additional dimension to the discussions on feasible and sustainable solutions to accelerate progress towards Millennium Development Goals 4 and 5, both at the global level but most importantly in individual countries. PMID- 18406864 TI - Every death counts: use of mortality audit data for decision making to save the lives of mothers, babies, and children in South Africa. AB - South Africa is one of the few developing countries with a national confidential inquiry into maternal deaths. 164 health facilities obtain audit data for stillbirths and neonatal deaths, and a new audit network does so for child deaths. Three separate reports have been published, providing valuable information about avoidable causes of death for mothers, babies, and children. These reports make health-system recommendations, many of which overlap and are intertwined with the scarcity of progress in addressing HIV/AIDS. The leaders of these three reports have united to prioritise actions to save the lives of South Africa's mothers, babies, and children. The country is off-track for the health related Millennium Development Goals. Mortality in children younger than 5 years has increased, whereas maternal and neonatal mortality remain constant. This situation indicates the challenge of strengthening the health system because of high inequity and HIV/AIDS. Coverage of services is fairly high, but addressing the gaps in quality and equity is essential to increasing the number of lives saved. Consistent leadership and accountability to address crosscutting health system and equity issues, and to prevent mother-to-child transmission of HIV, would save tens of thousands of lives every year. Audit is powerful, but only if the data lead to action. PMID- 18406866 TI - Of flukes and fistulae. PMID- 18406865 TI - Case management of HIV-infected severely malnourished children: challenges in the area of highest prevalence. PMID- 18406867 TI - Genomewide scan for loss of heterozygosity and chromosomal amplification in breast carcinoma using single-nucleotide polymorphism arrays. AB - In an effort to identify novel genes implicated in breast carcinogenesis, a genomewide scan for loss of heterozygosity (LOH) and copy number changes in paired-DNA samples extracted from normal and tumor tissue of frozen sections from women undergoing surgery for invasive breast cancer was conducted. The Affymetrix 10K SNP array was used to examine genomewide LOH of chromosomal regions. The number of LOH events, number of informative loci, percent heterozygosity, and percent fractional allelic loss (%FAL) were calculated. Although LOH events were detected in all samples, the proportion of LOH ranged from 0.1 to 57.2%. Elevated LOH events were detected in two samples, with a %FAL of 57.2 and 56.2. Chromosomal regions exceeding a threshold value for a P-value curve based on multiple-testing adjusted permutation methods were identified as significant regions of shared LOH across samples. Regions with significant LOH included 2p25.3; 2p21; 2p15 approximately p16.1, 2q23.3; and, 16q12.1. Chromosomal region 1q32.1 was identified as a region with significant copy number amplification. Regions of LOH and copy number changes identified from this analysis may provide insights into the underlying processes of and genes involved in breast carcinogenesis. The present study demonstrates a feasible methodological approach for the assessment of LOH and copy number changes. PMID- 18406868 TI - Genetic heterogeneity by comparative genomic hybridization in BRCAx breast cancers. AB - The chromosomal changes in eight familial BRCAx breast cancers (i.e., negative for BRCA1 or BRCA2) were analyzed by comparative genomic hybridization (CGH) to investigate intratumor heterogeneity. This was the first step in a study of most frequent chromosomal aberrations in BRCAx familial breast cancers. Laser microdissection analysis of paraffin tissue samples was followed by whole-genome amplification. CGH was performed on DNA isolated from two to three different cell groups per case to detect any cytogenetic aberrations in important clones that might have been missed when analyzing DNA extracted from large numbers of cells. The results were compared, to evaluate the influence of tumor heterogeneity on CGH, and the heterogeneity was confirmed comparing CGH with fluorescence in situ hybridization results. Different chromosomal aberrations were detected between adjacent clones within the same section, which highlights the utility of microdissection in addressing the problem of heterogeneity in whole-genome studies. Some chromosomal regions were more frequently altered in the eight BRCAx tumors; loss of 2q, 3p, 3q, 8p, 9p, and 15q and gains of 1p, 4p, 4q, 5p, 6q, 12q, and 19p were the most common. Further studies focusing on specific genes and sequences with more sensitive approaches, such as array-CGH, are warranted to confirm these findings. PMID- 18406869 TI - Screening for submicroscopic chromosomal rearrangements in Wilms tumor using whole-genome microarrays. AB - Wilms tumor is the fourth most common malignancy of childhood; its pathogenesis, however, remains largely unknown. With advancements in cytogenetic techniques, such as array comparative genomic hybridization (aCGH), there is new hope for uncovering small chromosomal microdeletions or microduplications that may contribute to our understanding of Wilms tumor. We performed aCGH on 10 samples of Wilms tumor with normal conventional cytogenetic and chromosomal CGH findings. Array CGH revealed abnormalities in 3 of the 10 samples, including microdeletions (2q37.1, 7q31 approximately q32, and 11q22.3), microduplication (18q21.1), and gains and losses of larger chromosomal areas (1q and 7q gain and loss of 7p, 11q, 14q, and 16q). Fluorescence in situ hybridization (FISH) analysis confirmed the abnormalities and revealed the majority of them existed only in a proportion cells (> or =30% of cells). We also performed aCGH on three samples of Wilms tumor with previously identified translocations between chromosomes 1 and 16, to determine the breakpoints. The breakpoints were seen in the pericentromeric regions of both chromosomes. Array CGH is useful for identifying submicroscopic changes in Wilms tumor and is more sensitive for detecting clonal abnormalities than conventional methods. PMID- 18406870 TI - Mechanisms of genesis of variant translocation in chronic myeloid leukemia are not correlated with ABL1 or BCR deletion status or response to imatinib therapy. AB - Many published studies have indicated that various mechanisms could be involved in the genesis of variant chronic myelogeneous leukemia (CML) translocations. These are mainly one-step or two-step mechanisms, associated or not with deletions adjacent to the translocation junction on der(9) or der(22) chromosomes (or both). Based on the mechanism of genesis, it has been suggested that the complexity may affect the occurrence of ABL1 and BCR deletions (either or both), or may be associated with the CML disease course, and thus could determine the response to imatinib therapy. Through a retrospective molecular cytogenetic study of 41 CML patients with variant Philadelphia chromosome (Ph), we explored the genesis of these variant rearrangements and analyzed the correlation with deletion status and imatinib efficiency. Our results confirmed that the one-step mechanism is the most frequent, evidenced in 30 of 41 patients (73%); 3 patients demonstrated other more complex multistep events and 8 patients (19.5%) harbored ABL1 or BCR deletions that are not significantly associated with the complexity of translocation genesis. We also found no association between one-step, two step, or multistep mechanisms and the response to imatinib therapy. PMID- 18406871 TI - Expression changes of CAV1 and EZH2, located on 7q31 approximately q36, are rarely related to genomic alterations in primary prostate carcinoma. AB - The chromosomal region 7q was repeatedly found to be rearranged in prostate carcinoma. It harbors several well described candidate tumor suppressor and oncogenes. We addressed two genes with opposite roles in cancer; CAV1, a putative tumor suppressor gene at 7q31, and EZH2 at 7q36, which is believed to promote tumor progression. Our primary aim was to assess their expression changes in primary tumors, and then to elucidate the underlying mechanism, assuming that genomic alterations of either locus could affect the other gene as well. In 35 prostate tumor samples, compared with adjacent tissues, CAV1 was overall downregulated (P < 10(-06)), whereas EZH2 was significantly overexpressed (P < 10(-06)). The observed dysregulations were coincident in nearly 70% of the cases. Copy number changes occurred in few tumors. Loss of CAV1 DNA was only marginally associated with reduced expression (P = 0.07), however, and genomic amplification of EZH2 could not explain its upregulation. Through bisulfite sequencing of four tumor samples, CpG-hypermethylation was verified as an alternative mechanism for CAV1 silencing, as reported previously. Moreover, it could also be involved in the reactivation of EZH2. PMID- 18406872 TI - Deletions adjacent to BCR and ABL1 breakpoints occur in a substantial minority of chronic myeloid leukemia patients with masked Philadelphia rearrangements. AB - Deletions at the t(9;22) breakpoint regions, found in 15% of chronic myeloid leukemia patients (CML) with an overt Philadelphia (Ph) translocation, are associated with an adverse disease prognosis in patients receiving interferon alpha therapy. The incidence of deletions has been shown to vary for different cytogenetic subgroups of CML, with a significantly higher incidence of deletion in patients with a variant Ph translocation. To date, however, the frequency of such deletions in the subgroup of CML patients in whom the BCR/ABL1 fusion arises via submicroscopic chromosomal insertion (masked Ph) has not been investigated. We report the evaluation of 14 patients with masked Ph-positive CML for the presence of deletions extending 3' from BCR and 5' from ABL1 using two triple color BCR/ABL probes. Deletions were identified in 3 patients (21%), encompassing sequences 5' to ABL1 in two of these and sequences 3' to BCR in the remaining patient, thus demonstrating that the phenomenon is a significant feature of the masked Ph CML subgroup. Furthermore, our findings are consistent with the notion that loss of genomic material is a potential side effect of any DNA breakage event at the 9q34.1 and 22q11.2 chromosomal regions, regardless of the subsequent mechanism of chromosomal rearrangement. PMID- 18406873 TI - Characterization of 9p21 copy number alterations in human melanoma by fluorescence in situ hybridization. AB - Alteration of the CDKN2A (alias p16) tumor suppressor gene, located on 9p21, occurs frequently in familial and sporadic melanomas. Beside CDKN2A, other genes (e.g., CDKN2B, and ARF/p14(ARF), long considered distinct from CDKN2A) on this locus are often deleted or mutated in a large number of tumors including glioma, bladder cancer, and lung cancer. The aim of this study was to evaluate the deletion pattern of the 9p21 locus on a cell-by-cell basis in a large number of melanoma samples using fluorescence in situ hybridization (FISH). In an analysis of 81 primary lesions targeting the 9p21 region and chromosome 9 centromere, high frequency of 9p21 loss (84%) was found. Deletion of 9p21 was present in both early- and late-stage melanomas with similar frequencies. Extra 9p21 copies were rarely seen; they were always associated with polysomy 9 and were observed only in advanced stage melanomas (6 tumors). This FISH study strengthens the hypothesis that the loss of 9p21 occurs frequently in primary melanoma, that the deletion is present in early and late stages of the disease with similar frequency, and that it affects a large extent of the locus. PMID- 18406874 TI - Spontaneous remission of myelodysplastic syndrome with monosomy 7 in a young boy. AB - Myelodysplastic syndrome (MDS) is a clonal hematopoietic disorder that often results in progression to acute myeloid leukemia (AML), particularly when additional genetic abnormalities are present, such as monosomy 7. Treatment options for this condition are limited. Spontaneous remission has occurred in a small number of cases. In the present case, a previously healthy 3-year-old boy diagnosed with MDS and monosomy 7 achieved spontaneous remission without intervention 30 months after initial diagnosis. Such findings highlight the need to monitor patients closely for evidence of spontaneous remission. PMID- 18406875 TI - Acute promyelocytic leukemia after mitoxantrone therapy for multiple sclerosis. AB - Mitoxantrone is a DNA-topoisomerase 2 inhibitor used as a single agent for treatment of relapsing-remitting or progressive multiple sclerosis (MS). We present here two patients treated with mitoxantrone for MS who subsequently developed acute promyelocytic leukemia (APL). These constitute, to our knowledge, the eighth and ninth reports of APL in patients treated with mitoxantrone for MS. Topoisomerase 2 inhibitors are associated with therapy-related acute myeloid leukemia (t-AML) with 11q23 abnormalities, but therapy-related APL (t-APL) is less common, and documentation of nine cases of t-APL after mitoxantrone therapy for MS suggests a specific association. PMID- 18406876 TI - Identification of a novel duplication in the APC gene using multiple ligation probe amplification in a patient with familial adenomatous polyposis. AB - Germline mutations in the adenomatous polyposis coli (APC) gene cause familial adenomatous polyposis (FAP), an autosomal dominant disease characterized by hundreds to thousands of adenomatous polyps in the colon and rectum, with progression to colorectal cancer. The majority of APC mutations are nucleotide substitutions and frameshift mutations that result in truncated proteins. Recently, large genomic alterations of the APC gene have been reported in FAP. DNA from 15 FAP patients, in whom no APC germline mutations were detected with denaturing high performance liquid chromatography, was analyzed with multiplex ligation-dependent probe amplification (MLPA) to evaluate gross genomic alterations in the APC gene. In one case, MLPA identified a novel duplication of exons 2-6 in one copy of the APC gene. Reverse transcriptase-polymerase chain reaction revealed that the mutant allele contained an in-frame multiexon duplication including 18 nucleotides located in exon 2, upstream of the ATG initiation codon. The presence of a premature stop codon in the duplicated sequence leads to the synthesis of a truncated APC polypeptide. These findings highlight the utility of evaluating infrequent APC mutation events in FAP patients using MLPA. PMID- 18406877 TI - A novel missense MSH2 gene mutation in a patient of a Korean family with hereditary nonpolyposis colorectal cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant cancer susceptible syndrome that predisposes to the early development of colorectal cancer. Germline mutations in DNA mismatch repair genes, particularly MLH1 and MSH2, are associated with the clinical phenotype of HNPCC. A previously unreported, novel missense mutation in exon 3 of the MSH2 gene (c.380A>T) was identified in the proband and a different missense mutation in exon 3 of MSH2 gene (c.505A>G) was noted in the mother, with a mutual splice mutation in intron 12 of the MSH2 gene in the proband, mother, and younger brother. Here, we report the clinical implications of a novel mutation in a patient with early-onset colorectal cancer and the significance of a common underlying splice site mutation occurring within a family with HNPCC. PMID- 18406878 TI - Cytogenetic analysis of a hemosiderotic fibrolipomatous tumor. AB - Hemosiderotic fibrolipomatous tumor is an extremely rare, nonencapsulated, fatty lesion with a consistent histologic appearance that was originally considered reactive in nature. To our knowledge, there are no previous reports on the cytogenetics of this lesion. Reported here is a case of hemosiderotic fibrolipomatous tumor arising within the subcutaneous tissue of the right foot, dorsal aspect, of an otherwise healthy 35-year-old woman. Subsequent cytogenetic analysis revealed a clonal reciprocal translocation between chromosomes 1 and 10, with a further rearrangement involving this derivative chromosome 1 and chromosome 3. This, in addition to its characteristic morphology and immunophenotype, supports the neoplastic nature of this tumor and may aid in its diagnosis. PMID- 18406879 TI - The GAS5 (growth arrest-specific transcript 5) gene fuses to BCL6 as a result of t(1;3)(q25;q27) in a patient with B-cell lymphoma. AB - The BCL6 gene is frequently disrupted at its 5' noncoding region by 3q27 chromosomal translocations in B-cell lymphoma. As a result of translocation, BCL6 is juxtaposed to reciprocal partners, such as the immunoglobulin (Ig) gene family. Besides the Ig loci, multiple non-Ig partners of the BCL6 translocation have been reported. Here we describe the identification of the GAS5 (growth arrest-specific transcript 5) gene as a novel partner of the BCL6 in a patient with diffuse large B-cell lymphoma, harboring the t(1;3)(q25;q27). In this case, the chromosome 1 breakpoint was located within the intronic small nucleolar RNA (snoRNA) sequence of GAS5 and the chromosome 3 breakpoint at 4 kb upstream of BCL6 exon 1a. As the result of chromosomal translocation, the GAS5-BCL6 chimeric transcripts were expressed, in which the 5'-terminal oligopyrimidine (5'TOP) sequence of GAS5 was fused to the whole coding sequence of BCL6. The GAS5 gene on chromosome 1q25 is the second BCL6 partner, to the SNHG5 on 6q15, which is classified as a non-protein-coding multiple snoRNA host and 5'-TOP class gene. PMID- 18406880 TI - Diabetes research and clinical practice: developing a leading medium for diabetes care, prevention and cure. PMID- 18406881 TI - Where do we draw the line? Contouring tumors on positron emission tomography/computed tomography. PMID- 18406882 TI - A novel MRI marker for prostate brachytherapy. AB - PURPOSE: Magnetic resonance imaging (MRI) is the optimal imaging modality for the prostate and surrounding critical organ structures. However, on MRI, the titanium radioactive seeds used for brachytherapy appear as black holes (negative contrast) and cannot be accurately localized. We sought to develop an encapsulated contrast agent marker (ECAM) with high-signal intensity on MRI to permit accurate localization of radioactive seeds with MRI during and after prostate brachytherapy. METHODS AND MATERIALS: We investigated several agents with paramagnetic and superparamagnetic properties. The agents were injected into titanium, acrylic, and glass seeds, which were linked together in various combinations and imaged with MRI. The agent with the greatest T1-weighted signal was tested further in a canine prostate and agarose phantom. Studies were performed on a 1.5-T clinical MRI scanner. RESULTS: The cobalt-chloride complex contrast (C4) agent with stoichiometry (CoCl(2))(0.8)(C(2)H(5)NO(2))(0.2) had the greatest T1-weighted signal (positive contrast) with a relaxivity ratio >1 (r(2)/r(1) = 1.21 +/- 0.29). Acrylic-titanium and glass-titanium seed strands were clearly visualized with the encapsulated contrast agent marker. CONCLUSION: We have developed a novel ECAM that permits positive identification of the radioactive seeds used for prostate brachytherapy on MRI. Preclinical in vitro phantom studies and in vivo canine studies are needed to further optimize MRI sequencing techniques to facilitate MRI-based dosimetry. PMID- 18406883 TI - Combined hypofractionated radiation and hormone therapy for the treatment of intermediate-risk prostate cancer. AB - PURPOSE: Because of the low alpha/beta value of prostate cancer, a therapeutic gain may be possible with a hypofractionated radiation scheme, and this gain may be further increased with the adjunct of hormone therapy. A Phase II study was undertaken to study the toxicity of such a treatment. METHODS AND MATERIALS: Forty-two patients with intermediate-risk prostate cancer were recruited for this study. Neoadjuvant and concomitant hormone therapy consisted of one injection of leuprolide acetate (4-month preparation) and 1 month of oral nonsteroidal, anti androgen medication starting on the day of the injection. Radiation treatment was started 8 weeks after the injection and patients received 57 Gy in 19 fractions. RESULTS: Median follow-up was 46 months. The treatment was well tolerated and no interruptions occurred. The majority (59%) had Grade 0 or 1 acute genitourinary (GU) toxicity, whereas 36% had Grade 2 and 5% had Grade 3 acute GU toxicity. Only Grade 1 or 2 gastrointestinal toxicity was seen. All chronic toxicity was of Grade 1 or 2 except for 3 patients (8%) with Grade 3 toxicity. Sixty-eight percent (68%) of patients had no long-term side effects from the treatment. At time of analysis, 79% showed no sign of treatment failure. CONCLUSIONS: Hypofractionated radiation with neoadjuvant and concomitant hormone therapy is well tolerated with no significant short- or long-term morbidity. Control for this risk group is good, and comparative Phase III studies should be undertaken to determine whether this treatment is superior to new evolving treatments. PMID- 18406884 TI - Relationship between neurocognitive function and quality of life after whole brain radiotherapy in patients with brain metastasis. AB - PURPOSE: To examine the relationship between neurocognitive function (NCF) and quality of life (QOL) in patients with brain metastases after whole-brain radiotherapy. PATIENTS AND METHODS: A total of 208 patients from the whole-brain radiotherapy arm of a Phase III trial (PCI-P120-9801), who underwent regular NCF and QOL (ADL [activities of daily living] and FACT-Br [Functional Assessment of Cancer Therapy-Brain-specific]) testing, were analyzed. Spearman's rank correlation was calculated between NCF and QOL, using each patient's own data, at each time point. To test the hypothesis that NCF declines before QOL changes, the predictive effect of NCF from previous visits on QOL was studied with a linear mixed-effects model. Neurocognitive function or QOL deterioration was defined relative to each patient's own baseline. Lead or lag time, defined as NCF deterioration before or after the date of QOL decline, respectively, was computed. RESULTS: At baseline, all NCF tests showed statistically significant correlations with ADL, which became stronger at 4 months. A similar observation was made with FACT-Br. Neurocognitive function scores from previous visits predicted ADL (p < 0.05 for seven of eight tests) or FACT-Br. Scores on all eight NCF tests deteriorated before ADL decline (net lead time 9-153 days); and scores on six of eight NCF tests deteriorated before FACT-Br (net lead time 9-82 days). CONCLUSIONS: Neurocognitive function and QOL are correlated. Neurocognitive function scores from previous visits are predictive of QOL. Neurocognitive function deterioration precedes QOL decline. The sequential association between NCF and QOL decline suggests that delaying NCF deterioration is a worthwhile treatment goal in brain metastases patients. PMID- 18406885 TI - A retrospective study of surgery and reirradiation for recurrent ependymoma. AB - PURPOSE: To report disease control for patients with recurrent ependymoma (EP) treated with surgery and a second course of radiation therapy (RT(2)). PATIENTS AND METHODS: Thirty-eight pediatric patients (median age, 2.7 years) with initially localized EP at the time of definitive RT underwent a second course of RT after local (n = 21), metastatic (n = 13), or combined (n = 4) failure. Reirradiation included radiosurgery (n = 6), focal fractionated reirradiation (n = 13), or craniospinal irradiation (CSI; n = 19). RESULTS: Initial time to failure was 16 months, and median age at second treatment was 4.8 years. Radiosurgery resulted in significant brainstem toxicity and one death (median dose, 18 Gy). Progression-free survival ratio was greater than unity for 4 of 6 patients; there was one long-term survivor. Three of 13 patients treated using focal fractionated reirradiation (median combined dose, 111.6 Gy) experienced metastasis. The CSI was administered to 12 patients with metastatic failure, 3 patients with local failure, and 4 patients with combined failure. The 4-year event-free survival rate was 53% +/- 20% for 12 patients with metastatic failure treated with CSI. Failure after CSI was observed in 1 of 3 patients with a history of local failure and 3 of 4 patients with a history of combined failure. CONCLUSION: Patients with locally recurrent EP experience durable local tumor control, but remain at risk of metastasis. Patients with metastatic EP failure may receive salvage therapy that includes a component of CSI. Durability of disease control and long-term effects from this approach require further follow up. PMID- 18406886 TI - Chemoradiation therapy for potentially resectable gastric cancer: clinical outcomes among patients who do not undergo planned surgery. AB - PURPOSE: We retrospectively analyzed treatment outcomes among resectable gastric cancer patients treated preoperatively with chemoradiation therapy (CRT) but rendered ineligible for planned surgery because of clinical deterioration or development of overt metastatic disease. METHODS AND MATERIALS: Between 1996 and 2004, 39 patients with potentially resectable gastric cancer received preoperative CRT but failed to undergo surgery. At baseline clinical staging, 33 (85%) patients had T3-T4 disease, and 27 (69%) patients had nodal involvement. Most patients received 45 Gy of radiotherapy with concurrent 5-fluorouracil-based chemotherapy. Twenty-one patients underwent induction chemotherapy before CRT. Actuarial times to local control (LC), distant control (DC), and overall survival (OS) were calculated by the Kaplan-Meier method. RESULTS: The cause for surgical ineligibility was development of metastatic disease (28 patients, 72%; predominantly peritoneal, 18 patients), poor performance status (5 patients, 13%), patient/physician preference (4 patients, 10%), and treatment-related death (2 patients, 5%). With a median follow-up of 8 months (range, 1-95 months), actuarial 1-year LC, DC, and OS were 46%, 12%, and 36%, respectively. Median LC and OS were 11.0 and 10.1 months, respectively. CONCLUSIONS: Patients with potentially resectable gastric cancer treated with preoperative CRT are found to be ineligible for surgery principally because of peritoneal progression. Patients who are unable to undergo planned surgery have outcomes comparable to that of patients with advanced gastric cancer treated with chemotherapy alone. CRT provides durable LC for the majority of the remaining life of these patients. PMID- 18406887 TI - Gene expression changes in cervical squamous cell carcinoma after initiation of chemoradiation and correlation with clinical outcome. AB - PURPOSE: The purpose of this study was to investigate early gene expression changes after chemoradiation in a human solid tumor, allowing identification of chemoradiation-induced gene expression changes in the tumor as well as the tumor microenvironment. In addition we aimed to identify a gene expression profile that was associated with clinical outcome. METHODS AND MATERIALS: Microarray experiments were performed on cervical cancer specimens obtained before and 48 h after chemoradiation from 12 patients with Stage IB2 to IIIB squamous cell carcinoma of the cervix treated between April 2001 and August 2002. RESULTS: A total of 262 genes were identified that were significantly changed after chemoradiation. Genes involved in DNA repair were identified including DDB2, ERCC4, GADD45A, and XPC. In addition, significantly regulated cell-to-cell signaling pathways included insulin-like growth factor-1 (IGF-1), interferon, and vascular endothelial growth factor signaling. At a median follow-up of 41 months, 5 of 12 patients had experienced either local or distant failure. Supervised clustering analysis identified a 58-gene set from the pretreatment samples that were differentially expressed between patients with and without recurrence. Genes involved in integrin signaling and apoptosis pathways were identified in this gene set. Immortalization-upregulated protein (IMUP), IGF-2, and ARHD had particularly marked differences in expression between patients with and without recurrence. CONCLUSIONS: Genetic profiling identified genes regulated by chemoradiation including DNA damage and cell-to-cell signaling pathways. Genes associated with recurrence were identified that will require validation in an independent patient data set to determine whether the 58-gene set associated with clinical outcome could be useful as a prognostic assay. PMID- 18406888 TI - Survivin antisense oligonucleotides effectively radiosensitize colorectal cancer cells in both tissue culture and murine xenograft models. AB - PURPOSE: Survivin shows a radiation resistance factor in colorectal cancer. In the present study, we determined whether survivin messenger RNA levels in patients with rectal cancer predict tumor response after neoadjuvant radiochemotherapy and whether inhibition of survivin by the use of antisense oligonucleotides (ASOs) enhances radiation responses. METHODS AND MATERIALS: SW480 colorectal carcinoma cells were transfected with survivin ASO (LY2181308) and irradiated with doses ranging from 0-8 Gy. Survivin expression, cell-cycle distribution, gammaH2AX fluorescence, and induction of apoptosis were monitored by means of immunoblotting, flow cytometry, and caspase 3/7 activity. Clonogenic survival was determined by using a colony-forming assay. An SW480 xenograft model was used to investigate the effect of survivin attenuation and irradiation on tumor growth. Furthermore, survivin messenger RNA levels were studied in patient biopsy specimens by using Affymetrix microarray analysis. RESULTS: In the translational study of 20 patients with rectal cancer, increased survivin levels were associated with significantly greater risk of local tumor recurrence (p = 0.009). Treatment of SW480 cells with survivin ASOs and irradiation resulted in an increased percentage of apoptotic cells, caspase 3/7 activity, fraction of cells in the G(2)/M phase, and H2AX phosphorylation. Clonogenic survival decreased compared with control-treated cells. Furthermore, treatment of SW480 xenografts with survivin ASOs and irradiation resulted in a significant delay in tumor growth. CONCLUSION: Survivin appears to be a molecular biomarker in patients with rectal cancer. Furthermore, in vitro and in vivo data suggest a potential role of survivin as a molecular target to improve treatment response to radiotherapy in patients with rectal cancer. PMID- 18406889 TI - Individual radiosensitivity measured with lymphocytes may predict the risk of acute reaction after radiotherapy. AB - PURPOSE: We tested whether the chromosomal radiosensitivity of in vitro irradiated lymphocytes could be used to predict the risk of acute reactions after radiotherapy. METHODS AND MATERIALS: Two prospective studies were performed: study A with 51 patients included different tumor sites and study B included 87 breast cancer patients. Acute reaction was assessed using the Radiation Therapy Oncology Group score. In both studies, patients were treated with curative radiotherapy, and the mean tumor dose applied was 55 Gy (40-65) +/- boost with 11 Gy (6-31) in study A and 50.4 Gy +/- boost with 10 Gy in study B. Individual radiosensitivity was determined with lymphocytes irradiated in vitro with X-ray doses of either 3 or 6 Gy and scoring the number of chromosomal deletions. RESULTS: Acute reactions displayed a typical spectrum with 57% in study A and 53% in study B showing an acute reaction of Grade 2-3. Individual radiosensitivity in both studies was characterized by a substantial variation and the fraction of patients with Grade 2-3 reaction was found to increase with increasing individual radiosensitivity measured at 6 Gy (study A, p = 0.238; study B, p = 0.023). For study B, this fraction increased with breast volume, and the impact of individual radiosensitivity on acute reaction was especially pronounced (p = 0.00025) for lower breast volume. No such clear association with acute reaction was observed when individual radiosensitivity was assessed at 3 Gy. CONCLUSION: Individual radiosensitivity determined at 6 Gy seems to be a good predictor for risk of acute effects after curative radiotherapy. PMID- 18406890 TI - Examining margin reduction and its impact on dose distribution for prostate cancer patients undergoing daily cone-beam computed tomography. AB - PURPOSE: To examine the dosimetric impact of margin reduction and quantify residual error after three-dimensional (3D) image registration using daily cone beam computed tomography (CBCT) for prostate cancer patients. METHODS AND MATERIALS: One hundred forty CBCTs from 5 prostate cancer patients were examined. Two intensity-modulated radiotherapy plans were generated on CT simulation on the basis of two planning target volume (PTV) margins: 10 mm all around the prostate and seminal vesicles except 6 mm posteriorly (10/6) and 5 mm all around except 3 mm posteriorly (5/3). Daily CBCT using the Varian On-Board Imaging System was acquired. The 10/6 and 5/3 simulation plans were overlaid onto each CBCT, and each CBCT plan was calculated. To examine residual error, PlanCT/CBCT intensity based 3D image registration was performed for prostate localization using center of mass and maximal border displacement. RESULTS: Prostate coverage was within 2% between the 10/6 and 5/3 plans. Seminal vesicle coverage was reduced with the 5/3 plan compared with the 10/6 plan, with coverage difference within 7%. The 5/3 plan allowed 30-50% sparing of bladder and rectal high-dose regions. For residual error quantification, center of mass data show that 99%, 93%, and 96% of observations fall within 3 mm in the left-right, anterior-posterior, and superior inferior directions, respectively. Maximal border displacement observations range from 79% to 99%, within 5 mm for all directions. CONCLUSION: Cone-beam CT dosimetrically validated a 10/6 margin when soft-tissue localization is not used. Intensity-based 3D image registration has the potential to improve target localization and to provide guidelines for margin definition. PMID- 18406891 TI - Daily bone alignment with limited repeat CT correction rivals daily ultrasound alignment for prostate radiotherapy. AB - PURPOSE: To compare the effectiveness of daily ultrasound (US)- and computed tomography (CT)-guided alignments with an off-line correction protocol using daily bone alignment plus a correction factor for systematic internal prostate displacement (CF(ID)). METHODS AND MATERIALS: Ten prostate cancer patients underwent CT scans three times weekly using an integrated CT-linear accelerator system, followed by alignment using US for daily radiotherapy. Intensity modulated radiotherapy plans were designed with our current clinical margins. The treatment plan was copied onto the repeat CT images and aligned using several methods: (1) bone alignment plus CF(ID) after three off-line CT scans (bone+3CT), (2) bone alignment plus CF(ID) after six off-line CT scans (bone+6CT), (3) US alignment, and (4) CT alignment. The accuracy of the repeated US and CT measurements to determine the CF(ID) was compared. The target dosimetric effect was quantified. RESULTS: The CF(ID) for internal systematic prostate displacements was more accurately measured with limited repeat CT scans than with US (residual error, 0.0 +/- 0.7 mm vs. 2.0 +/- 3.2 mm). Bone+3CT, bone+6CT, and US provided equivalent prostate and seminal vesicle dose coverage, but bone+3CT and bone+6CT produced more precise daily alignments. Daily CT alignment provided the greatest target dose coverage. CONCLUSION: Daily bone alignment plus CF(ID) for internal systematic prostate displacement provided better daily alignment precision and equivalent dose coverage compared with daily US alignment. The CF(ID) should be based on at least three repeat CT scans, which could be collected before the start of treatment or during the first 3 treatment days. Daily bone alignment plus CF(ID) provides another option for accurate prostate cancer patient positioning. PMID- 18406892 TI - Organ deformation and dose coverage in robotic respiratory-tracking radiotherapy. AB - PURPOSE: Respiratory motion presents a significant challenge in stereotactic body radiosurgery. Respiratory tracking that follows the translational movement of the internal fiducials minimizes the uncertainties in dose delivery. However, the effect of deformation, defined as any changes in the body and organs relative to the center of fiducials, remains unanswered. This study investigated this problem and a possible solution. METHODS AND MATERIALS: Dose delivery using a robotic respiratory-tracking system was studied with clinical data. Each treatment plan was designed with the computed tomography scan in the end-expiration phase. The planned beams were applied to the computed tomography scan in end-inspiration following the shift of the fiducials. The dose coverage was compared with the initial plan, and the uncertainty due to the deformation was estimated. A necessary margin from the clinical target volume to the planning target volume was determined to account for this and other sources of uncertainty. RESULTS: We studied 12 lung and 5 upper abdomen lesions. Our results demonstrated that for lung patients with properly implanted fiducials a 3-mm margin is required to compensate for the deformation and a 5-mm margin is required to compensate for all uncertainties. Our results for the upper abdomen tumors were still preliminary but indicated a similar result, although a larger margin might be required. CONCLUSION: The effect of body deformation was studied. We found that adequate dose coverage for lung tumors can be ensured with proper fiducial placement and a 5-mm planning target volume margin. This approach is more practical and effective than a recent proposal to combine four-dimensional planning with respiratory tracking. PMID- 18406893 TI - Comparison of rigid and adaptive methods of propagating gross tumor volume through respiratory phases of four-dimensional computed tomography image data set. AB - PURPOSE: To compare three different methods of propagating the gross tumor volume (GTV) through the respiratory phases that constitute a four-dimensional computed tomography image data set. METHODS AND MATERIALS: Four-dimensional computed tomography data sets of 20 patients who had undergone definitive hypofractionated radiotherapy to the lung were acquired. The GTV regions of interest (ROIs) were manually delineated on each phase of the four-dimensional computed tomography data set. The ROI from the end-expiration phase was propagated to the remaining nine phases of respiration using the following three techniques: (1) rigid-image registration using in-house software, (2) rigid image registration using research software from a commercial radiotherapy planning system vendor, and (3) rigid image registration followed by deformable adaptation originally intended for organ-at-risk delineation using the same software. The internal GTVs generated from the various propagation methods were compared with the manual internal GTV using the normalized Dice similarity coefficient (DSC) index. RESULTS: The normalized DSC index of 1.01 +/- 0.06 (SD) for rigid propagation using the in house software program was identical to the normalized DSC index of 1.01 +/- 0.06 for rigid propagation achieved with the vendor's research software. Adaptive propagation yielded poorer results, with a normalized DSC index of 0.89 +/- 0.10 (paired t test, p <0.001). CONCLUSION: Propagation of the GTV ROIs through the respiratory phases using rigid- body registration is an acceptable method within a 1-mm margin of uncertainty. The adaptive organ-at-risk propagation method was not applicable to propagating GTV ROIs, resulting in an unacceptable reduction of the volume and distortion of the ROIs. PMID- 18406894 TI - Image-guided radiotherapy for liver cancer using respiratory-correlated computed tomography and cone-beam computed tomography. AB - PURPOSE: To evaluate a novel four-dimensional (4D) image-guided radiotherapy (IGRT) technique in stereotactic body RT for liver tumors. METHODS AND MATERIALS: For 11 patients with 13 intrahepatic tumors, a respiratory-correlated 4D computed tomography (CT) scan was acquired at treatment planning. The target was defined using CT series reconstructed at end-inhalation and end-exhalation. The liver was delineated on these two CT series and served as a reference for image guidance. A cone-beam CT scan was acquired after patient positioning; the blurred diaphragm dome was interpreted as a probability density function showing the motion range of the liver. Manual contour matching of the liver structures from the planning 4D CT scan with the cone-beam CT scan was performed. Inter- and intrafractional uncertainties of target position and motion range were evaluated, and interobserver variability of the 4D-IGRT technique was tested. RESULTS: The workflow of 4D-IGRT was successfully practiced in all patients. The absolute error in the liver position and error in relation to the bony anatomy was 8 +/- 4 mm and 5 +/- 2 mm (three-dimensional vector), respectively. Margins of 4-6 mm were calculated for compensation of the intrafractional drifts of the liver. The motion range of the diaphragm dome was reproducible within 5 mm for 11 of 13 lesions, and the interobserver variability of the 4D-IGRT technique was small (standard deviation, 1.5 mm). In 4 patients, the position of the intrahepatic lesion was directly verified using a mobile in-room CT scanner after application of intravenous contrast. CONCLUSION: The results of our study have shown that 4D image guidance using liver contour matching between respiratory-correlated CT and cone-beam CT scans increased the accuracy compared with stereotactic positioning and compared with IGRT without consideration of breathing motion. PMID- 18406895 TI - Investigation of interfraction variations of MammoSite balloon applicator in high dose-rate brachytherapy of partial breast irradiation. AB - PURPOSE: To measure the interfraction changes of the MammoSite applicator and evaluate their dosimetric effect on target coverage and sparing of organs at risk. METHODS AND MATERIALS: A retrospective evaluation of the data from 19 patients who received 10 fractions (34 Gy) of high-dose-rate partial breast irradiation was performed. A computed tomography-based treatment plan was generated for Fraction 1, and a computed tomography scan was acquired just before the delivery of each fraction to ensure a consistent shape of the balloon. The eccentricity, asymmetry, and planning target volume (PTV) for plan evaluation purposes (PTV_EVAL), as well as trapped air gaps, were measured for all patients. Furthermore, 169 computed tomography-based treatment plans were retrospectively generated for Fractions 2-10. Interfraction dosimetric variations were evaluated using the %PTV_EVAL coverage, target dose homogeneity index, target dose conformal index, and maximum doses to the organs at risks. RESULTS: The average variation of eccentricity and asymmetry from Fraction 1 values of 3.5% and 1.1 mm was -0.4% +/- 1.6% and -0.1 +/- 0.6 mm. The average trapped air gap volume was dramatically reduced from before treatment (3.7 cm(3)) to Fraction 1 (0.8 cm(3)). The PTV_EVAL volume change was insignificant. The average variation for the %PTV_EVAL, target dose homogeneity, and target dose conformal index from Fraction 1 values of 94.7%, 0.64, and 0.85 was 0.15% +/- 2.4%, -0.35 +/- 2.4%, and -0.34 +/- 4.9%, respectively. The average Fraction 1 maximum skin and ipsilateral lung dose of 3.2 Gy and 2.0 Gy varied by 0.08 +/- 0.47 and -0.16 +/- 0.29 Gy, respectively. CONCLUSION: The interfraction variations were patient specific and fraction dependent. Although the average interfraction dose variations for the target and organs at risk were not clinically significant, the maximum variations could be clinically significant. PMID- 18406897 TI - Updated analysis of RTOG 94-13: in regard to Lawton et al. (Int J Radiat Oncol Biol Phys 2007;69:646-655). PMID- 18406899 TI - Transperineal injection of hyaluronic acid in anterior perirectal fat to decrease rectal toxicity from radiation delivered with intensity-modulated brachytherapy or EBRT for prostate cancer patients: in regard to Prada et al. (Int J Radiat Oncol Biol Phys 2007;69:95-102.). PMID- 18406900 TI - An update of the phase III trial comparing whole pelvic to prostate only radiotherapy and neoadjuvant to adjuvant total androgen suppression: updated analysis of RTOG 94-13, with emphasis on unexpected hormone/radiation interactions: in regard to Lawton et al. (Int J Radiat Oncol Biol Phys 2007;69:646-655.). PMID- 18406902 TI - In regard to Chen et Al. (Int J Radiat Oncol Biol Phys 2007;69:25-31) and Chao et Al. (Int j Radiat Oncol Biol Phys 2007;69:32-40). PMID- 18406904 TI - Foreword. Symposium " Quality Assurance of Radiation Therapy: The Challenges of Advanced Technologies". PMID- 18406905 TI - Quality assurance challenges for motion-adaptive radiation therapy: gating, breath holding, and four-dimensional computed tomography. AB - Compared with conventional three-dimensional (3D) conformal radiation therapy and intensity-modulated radiation therapy treatments, quality assurance (QA) for motion-adaptive radiation therapy involves various challenges because of the added temporal dimension. Here we discuss those challenges for three specific techniques related to motion-adaptive therapy: namely respiratory gating, breath holding, and four-dimensional computed tomography. Similar to the introduction of any other new technologies in clinical practice, typical QA measures should be taken for these techniques also, including initial testing of equipment and clinical procedures, as well as frequent QA examinations during the early stage of implementation. Here, rather than covering every QA aspect in depth, we focus on some major QA challenges. The biggest QA challenge for gating and breath holding is how to ensure treatment accuracy when internal target position is predicted using external surrogates. Recommended QA measures for each component of treatment, including simulation, planning, patient positioning, and treatment delivery and verification, are discussed. For four-dimensional computed tomography, some major QA challenges have also been discussed. PMID- 18406906 TI - Quality assurance of intensity-modulated radiation therapy. AB - The current paradigm for the quality assurance (QA) program for intensity modulated radiation therapy (IMRT) includes QA of the treatment planning system, QA of the delivery system, and patient-specific QA. Although the IMRT treatment planning and delivery system is the same as for conventional three-dimensional conformal radiation therapy, it has more parameters to coordinate and verify. Because of complex beam intensity modulation, each IMRT field often includes many small irregular off-axis fields, resulting in isodose distributions for each IMRT plan that are more conformal than those from conventional treatment plans. Therefore, these features impose a new and more stringent set of QA requirements for IMRT planning and delivery. The generic test procedures to validate dose calculation and delivery accuracy for both treatment planning and IMRT delivery have to be customized for each type of IMRT planning and delivery strategy. The rationale for such an approach is that the overall accuracy of IMRT delivery is incumbent on the piecewise uncertainties in both the planning and delivery processes. The end user must have well-defined evaluation criteria for each element of the planning and delivery process. Such information can potentially be used to determine a priori the accuracy of IMRT planning and delivery. PMID- 18406907 TI - Helical tomotherapy quality assurance. AB - Helical tomotherapy uses a dynamic delivery in which the gantry, treatment couch, and multileaf collimator leaves are all in motion during treatment. This results in highly conformal radiotherapy, but the complexity of the delivery is partially hidden from the end-user because of the extensive integration and automation of the tomotherapy control systems. This presents a challenge to the medical physicist who is expected to be both a system user and an expert, capable of verifying relevant aspects of treatment delivery. A related issue is that a clinical tomotherapy planning system arrives at a customer's site already commissioned by the manufacturer, not by the clinical physicist. The clinical physicist and the manufacturer's representative verify the commissioning at the customer site before acceptance. Theoretically, treatment could begin immediately after acceptance. However, the clinical physicist is responsible for the safe and proper use of the machine. In addition, the therapists and radiation oncologists need to understand the important machine characteristics before treatment can proceed. Typically, treatment begins about 2 weeks after acceptance. This report presents an overview of the tomotherapy system. Helical tomotherapy has unique dosimetry characteristics, and some of those features are emphasized. The integrated treatment planning, delivery, and patient-plan quality assurance process is described. A quality assurance protocol is proposed, with an emphasis on what a clinical medical physicist could and should check. Additionally, aspects of a tomotherapy quality assurance program that could be checked automatically and remotely because of its inherent imaging system and integrated database are discussed. PMID- 18406908 TI - Linear accelerator and gamma knife-based stereotactic cranial radiosurgery: challenges and successes of existing quality assurance guidelines and paradigms. AB - Intracranial stereotactic radiosurgery has been practiced since 1951. The technique has expanded from a single dedicated unit in Stockholm in 1968 to hundreds of centers performing an estimated 100,000 Gamma Knife and linear accelerator cases in 2005. The radiation dosimetry of small photon fields used in this technique has been well explored in the past 15 years. Quality assurance recommendations have been promulgated in refereed reports and by several national and international professional societies since 1991. The field has survived several reported treatment errors and incidents, generally reacting by strengthening standards and precautions. An increasing number of computer controlled and robotic-dedicated treatment units are expanding the field and putting patients at risk of unforeseen errors. Revisions and updates to previously published quality assurance documents, and especially to radiation dosimetry protocols, are now needed to ensure continued successful procedures that minimize the risk of serious errors. PMID- 18406909 TI - Quality assurance procedures for stereotactic body radiation therapy. AB - Cranial stereotactic radiosurgery (SRS) and radiotherapy (SRT) are established treatment modalities. Initial implementations of these techniques rigidly attached frames to the patient's head for single-fraction treatments. The head frame accommodates an external fiducial marker system that is a reliable reference for targets within the cranium and accurately links the imaging equipment used for treatment planning to the treatment device. Fractionated SRT treatments use noninvasive "relocatable"-type head immobilization that fixes to the patient's head and face features. Clearly defined quality assurance (QA) procedures exist for both cranial SRS and SRT but are not as well developed for extracranial SRT. Procedures for demonstrating the geometric relationship between the planning imaging and treatment have to some degree copied the techniques used for intracranial stereotactic irradiation. However, there are some unique QA issues that are specific to extracranial irradiation. One major consideration is the large number of methodologies available for stereotactic body radiation therapy. In addition to the variety of integrated image-guided frameless systems, there are immobilization devices (called body frame systems) that use a fiducial reference system similar to the cranial devices. This article describes generic QA approaches that can be adapted to the various stereotactic body radiation therapy methodologies. PMID- 18406910 TI - Cyberknife image-guided delivery and quality assurance. AB - The CyberKnife is a complex, emerging technology that is a significant departure from current stereotactic radiosurgery and external beam radiotherapy technologies. In its clinical application and quality assurance (QA) approach, the CyberKnife is currently situated somewhere in between stereotactic radiosurgery and radiotherapy. The clinical QA for this image-guided treatment delivery system typically follows the vendor's guidance, mainly because of the current lack of vendor-independent QA recommendations. The problem has been exacerbated because very little published data are available for QA for the CyberKnife system, especially for QA of the interaction between individual system components. The tools and techniques for QA of the CyberKnife are under development and will continue to improve with longer clinical experience of the users. The technology itself continues to evolve, forcing continuous changes and adaptation of QA. To aid in the process of developing comprehensive guidance on CyberKnife QA, a database of errors based on users reporting incidents and corrective actions would be desirable. The goal of this work was to discuss the status of QA guidelines in the clinical implementation of the CyberKnife system. This investigation was done from the perspective of an active clinical and research site using the CyberKnife. PMID- 18406911 TI - Current external beam radiation therapy quality assurance guidance: does it meet the challenges of emerging image-guided technologies? AB - The traditional prescriptive quality assurance (QA) programs that attempt to ensure the safety and reliability of traditional external beam radiation therapy are limited in their applicability to such advanced radiation therapy techniques as three-dimensional conformal radiation therapy, intensity-modulated radiation therapy, inverse treatment planning, stereotactic radiosurgery/radiotherapy, and image-guided radiation therapy. The conventional QA paradigm, illustrated by the American Association of Physicists in Medicine Radiation Therapy Committee Task Group 40 (TG-40) report, consists of developing a consensus menu of tests and device performance specifications from a generic process model that is assumed to apply to all clinical applications of the device. The complexity, variation in practice patterns, and level of automation of high-technology radiotherapy renders this "one-size-fits-all" prescriptive QA paradigm ineffective or cost prohibitive if the high-probability error pathways of all possible clinical applications of the device are to be covered. The current approaches to developing comprehensive prescriptive QA protocols can be prohibitively time consuming and cost ineffective and may sometimes fail to adequately safeguard patients. It therefore is important to evaluate more formal error mitigation and process analysis methods of industrial engineering to more optimally focus available QA resources on process components that have a significant likelihood of compromising patient safety or treatment outcomes. PMID- 18406912 TI - Quality assurance of immobilization and target localization systems for frameless stereotactic cranial and extracranial hypofractionated radiotherapy. AB - The success of stereotactic radiosurgery has stimulated significant interest in the application of such an approach for the treatment of extracranial tumors. The potential benefits of reduced healthcare costs and improved patient outcomes that could be realized in a high-precision, hypofractionated treatment paradigm are numerous. Image-guidance technologies are eliminating the historic requirement for rigid head fixation and will also accelerate the clinical implementation of the approach in extracranial sites. An essential prerequisite of "frameless" stereotactic systems is that they provide localization accuracy consistent with the safe delivery of a therapeutic radiation dose given in one or few fractions. In this report, we reviewed the technologies for frameless localization of cranial and extracranial targets with emphasis on the quality assurance aspects. PMID- 18406913 TI - Quality assurance issues for computed tomography-, ultrasound-, and magnetic resonance imaging-guided brachytherapy. AB - The requirements of quality assurance (QA) for both brachytherapy and imaging devices are well-defined, but image-guided brachytherapy has raised new issues. Image guidance in brachytherapy involves the transition from reference point dosimetry using films to volumetric imaging such as computed tomography, ultrasonography, and magnetic resonance imaging for treatment planning and guidance of applicator, needle, or seed placement. The QA of these devices might not reflect the conditions of use in brachytherapy or the requirements of brachytherapy treatment planning. Image interpretation becomes much more important with image-guided brachytherapy. The success of a procedure could depend on the interpretation of a single image in a calibration phase done under the time pressures of the operative setting. This change has implications at the level of treatment, the process, and the field of brachytherapy as a whole. The QA concerns arising from brachytherapy procedures using ultrasound, computed tomography, and magnetic resonance imaging guidance are discussed, as are the problems associated with using imaging in an interventional setting. This report was intended to indicate the QA concerns arising from the convergence of brachytherapy and imaging-highlighting areas in which technical improvements are needed. PMID- 18406914 TI - Clinical practice and quality assurance challenges in modern brachytherapy sources and dosimetry. AB - Modern brachytherapy has led to effective treatments through the establishment of broadly applicable dosimetric thresholds for maximizing survival with minimal morbidity. Proper implementation of recent dosimetric consensus statements and quality assurance procedures is necessary to maintain the established level of safety and efficacy. This review classifies issues as either "systematic" or "stochastic" in terms of their impact on large groups or individual patients, respectively. Systematic changes affecting large numbers of patients occur infrequently and include changes in source dosimetric parameters, prescribing practice, dose calculation formalism, and improvements in calculation algorithms. The physicist must be aware of how incipient changes accord with previous experience. Stochastic issues involve procedures that are applied to each patient individually. Although ample guidance for quality assurance of brachytherapy sources exists, some ambiguities remain. The latest American Association of Physicists in Medicine guidance clarifies what is meant by independent assay, changes source sampling recommendations, particularly for sources in sterile strands and sterile preassembled needles, and modifies action level thresholds. The changing environment of brachytherapy has not changed the fact that the prime responsibility for quality assurance in brachytherapy lies with the institutional medical physicist. PMID- 18406915 TI - Quality assurance issues for therapeutic application of radioactive microspheres. AB - The use of radioactive microspheres for the treatment of hepatic cancer is a procedure that raises unique quality assurance (QA) concerns. The greatest of these concerns is the coordination of the responsibilities among the medical team members from interventional radiology, radiation oncology, nuclear medicine, and medical physics. A single QA practice and procedure guidance document does not currently exist that addresses the range of issues of concern for radioactive microspheres. A small sampling of QA issues of concern include imaging QA, procedure-specific imaging protocols, detector calibration, activity measurement, radiation safety, patient dose calculations, and patient-specific QA. Some of the items listed have historically been the responsibility of a single team member, and other items have been concerns for all. A procedural overview of the therapeutic application of radioactive microspheres is presented to illustrate the broad, team-based QA approach necessary to safely and effectively deliver this type of treatment. From this overview, the reader will be able to customize the local QA protocol to meet the local division of responsibilities. PMID- 18406916 TI - Quality assurance/quality control issues for intraoperative planning and adaptive repeat planning of image-guided prostate implants. AB - The quality assurance/quality control purpose is this. We design a treatment plan, and we wish to be as certain as reasonably possible that the treatment is delivered as planned. In the case of conventionally planned prostate brachytherapy, implementing to the letter the implantation plan is rarely attainable and therefore can require adaptive replanning (a quality control issue). The reasons for this state of affairs include changes in the prostate shape and volume during implantation and treatment delivery (e.g., edema resolution) and unavoidable inaccuracy in the placement of the seeds in the prostate. As a result, quality-control activities (e.g., the need to monitor ideally, on the fly-the target and urethral and rectal dosage) must be also addressed. PMID- 18406917 TI - Safety and accountability in healthcare from past to present. AB - Healthcare is transitioning into a new era-an era of accountability. This era demands heightened awareness of the quality, cost, and safety of healthcare, with value (quality/cost) and safety being the watchwords of accountability. Many factors are driving this transition, and it is affecting all healthcare disciplines, including radiation oncology. The transition is accompanied by the transformation of healthcare from a craft-based culture to an information-age culture in which patient needs and information are given top priority. These changes call for new measures to quantify and document the value and safety of procedures in radiation oncology. PMID- 18406918 TI - Errors in radiotherapy: motivation for development of new radiotherapy quality assurance paradigms. AB - Modern radiotherapy practice has rapidly evolved during the past decade, making use of many highly complex and/or automated processes for planning and delivery, including new techniques, like intensity-modulated radiotherapy driven by inverse planning optimization methods, or near real-time image-guided adaptive therapy based on fluoroscopic or tomographic imaging on the treatment machine. In spite of the modern technology, or potentially because of it in some instances, errors and other problems continue to have a significant impact on the field. This report reviews example errors and problems, discusses some of the quality assurance issues that these types of problems raise, and motivates the development of more modern and sophisticated approaches to assure quality for our clinical radiotherapy treatment methods. PMID- 18406919 TI - Critique of traditional quality assurance paradigm. AB - Radiotherapy was one of the first medical specialties to incorporate quality assurance (QA) into routine practices. Over the years, QA has become more comprehensive and time consuming. Following all the recommended QA procedures would burden the resources of most physics staffs. Much of the QA in recommendations serves no effective role. Performing all appropriate QA for the treatment units does not necessarily prevent events that result in patient injury. Equipment failures that have produced injury often occur with no warning. The human reactions to such failures will determine whether injury will follow. In addition, most events occur, not because of equipment failures, but because of human actions. Effective QA needs to consider procedures, rather than just equipment, and the human actions and interactions required for safe execution. New approaches address this problem. PMID- 18406920 TI - A method for evaluating quality assurance needs in radiation therapy. AB - The increasing complexity of modern radiation therapy planning and delivery techniques challenges traditional prescriptive quality control and quality assurance programs that ensure safety and reliability of treatment planning and delivery systems under all clinical scenarios. Until now quality management (QM) guidelines published by concerned organizations (e.g., American Association of Physicists in Medicine [AAPM], European Society for Therapeutic Radiology and Oncology [ESTRO], International Atomic Energy Agency [IAEA]) have focused on monitoring functional performance of radiotherapy equipment by measurable parameters, with tolerances set at strict but achievable values. In the modern environment, however, the number and sophistication of possible tests and measurements have increased dramatically. There is a need to prioritize QM activities in a way that will strike a balance between being reasonably achievable and optimally beneficial to patients. A systematic understanding of possible errors over the course of a radiation therapy treatment and the potential clinical impact of each is needed to direct limited resources in such a way to produce maximal benefit to the quality of patient care. Task Group 100 of the AAPM has taken a broad view of these issues and is developing a framework for designing QM activities, and hence allocating resources, based on estimates of clinical outcome, risk assessment, and failure modes. The report will provide guidelines on risk assessment approaches with emphasis on failure mode and effect analysis (FMEA) and an achievable QM program based on risk analysis. Examples of FMEA to intensity-modulated radiation therapy and high-dose-rate brachytherapy are presented. Recommendations on how to apply this new approach to individual clinics and further research and development will also be discussed. PMID- 18406921 TI - Human factors and systems engineering approach to patient safety for radiotherapy. AB - The traditional approach to solving patient safety problems in healthcare is to blame the last person to touch the patient. But since the publication of To Err is Human, the call has been instead to use human factors and systems engineering methods and principles to solve patient safety problems. However, an understanding of the human factors and systems engineering is lacking, and confusion remains about what it means to apply their principles. This paper provides a primer on them and their applications to patient safety. PMID- 18406922 TI - Error prevention as developed in airlines. AB - The airline industry is a high-risk endeavor. Tens of thousands of flights depart each day carrying millions of passengers with the potential for catastrophic consequences. To manage and mitigate this risk, airline operators, labor unions, and the Federal Aviation Administration have developed a partnership approach to improving safety. This partnership includes cooperative programs such as the Aviation Safety Action Partnership and the Flight Operational Quality Assurance. It also involves concentrating on the key aspects of aircraft maintenance reliability and employee training. This report discusses recent enhancements within the airline industry in the areas of proactive safety programs and the move toward safety management systems that will drive improvements in the future. PMID- 18406923 TI - Current brachytherapy quality assurance guidance: does it meet the challenges of emerging image-guided technologies? AB - In the past decade, brachytherapy has shifted from the traditional surgical paradigm to more modern three-dimensional image-based planning and delivery approaches. The role of intraoperative and multimodality image-based planning is growing. Published American Association of Physicists in Medicine, American College of Radiology, European Society for Therapeutic Radiology and Oncology, and International Atomic Energy Agency quality assurance (QA) guidelines largely emphasize the QA of planning and delivery devices rather than processes. These protocols have been designed to verify compliance with major performance specifications and are not risk based. With some exceptions, complete and clinically practical guidance exists for sources, QA instrumentation, non-image based planning systems, applicators, remote afterloading systems, dosimetry, and calibration. Updated guidance is needed for intraoperative imaging systems and image-based planning systems. For non-image-based brachytherapy, the American Association of Physicists in Medicine Task Group reports 56 and 59 provide reasonable guidance on procedure-specific process flow and QA. However, improved guidance is needed even for established procedures such as ultrasound-guided prostate implants. Adaptive replanning in brachytherapy faces unsolved problems similar to that of image-guided adaptive external beam radiotherapy. PMID- 18406924 TI - Improving patient safety in anesthesia: a success story? AB - Anesthesia is necessary for surgery; however, it does not deliver any direct therapeutic benefit. The risks of anesthesia must therefore be as low as possible. Anesthesiology has been identified as a leader in improving patient safety. Anesthetic mortality has decreased, and in healthy patients can be as low as 1:250,000. Trends in anesthetic morbidity have not been as well defined, but it appears that the risk of injury is decreasing. Studies of error during anesthesia and Closed Claims studies have identified sources of risk and methods to reduce the risks associated with anesthesia. These include changes in technology, such as anesthetic delivery systems and monitors, the application of human factors, the use of simulation, and the establishment of reporting systems. A review of the important events in the past 50 years illustrates the many steps that have contributed to the improvements in anesthesia safety. PMID- 18406925 TI - Tools for developing a quality management program: proactive tools (process mapping, value stream mapping, fault tree analysis, and failure mode and effects analysis). AB - This article examines the concepts of quality management (QM) and quality assurance (QA), as well as the current state of QM and QA practices in radiotherapy. A systematic approach incorporating a series of industrial engineering-based tools is proposed, which can be applied in health care organizations proactively to improve process outcomes, reduce risk and/or improve patient safety, improve through-put, and reduce cost. This tool set includes process mapping and process flowcharting, failure modes and effects analysis (FMEA), value stream mapping, and fault tree analysis (FTA). Many health care organizations do not have experience in applying these tools and therefore do not understand how and when to use them. As a result there are many misconceptions about how to use these tools, and they are often incorrectly applied. This article describes these industrial engineering-based tools and also how to use them, when they should be used (and not used), and the intended purposes for their use. In addition the strengths and weaknesses of each of these tools are described, and examples are given to demonstrate the application of these tools in health care settings. PMID- 18406926 TI - Tools for developing a quality management program: human factors and systems engineering tools. AB - During the past 10 years, there has been growing acceptance and encouragement of partnerships between medical teams and engineers. Using human factors and systems engineering descriptions of process flows and operational sequences, the author's research laboratory has helped highlight opportunities for reducing adverse events and improving performance in health care and other high-consequence environments. This research emphasized studying human behavior that enhances system performance and a range of factors affecting adverse events, rather than a sole emphasis on human error causation. Developing a balanced evaluation requires novel approaches to causal analyses of adverse events and, more importantly, methods of recovery from adverse conditions. Recent work by the author's laboratory in collaboration with the Regenstrief Center for Healthcare Engineering has started to address possible improvements in taxonomies describing health care tasks. One major finding includes enhanced understanding of events and how event dynamics influence provider tasks and constraints. Another element of this research examines team coordination tasks that strongly affect patient care and quality management, but may be undervalued as "indirect patient care" activities. PMID- 18406927 TI - Linking quality assurance to performance improvement to produce a high reliability organization. AB - Three basic change management models are currently used in healthcare to produce and sustain quality improvement. We have presented the context to determine where any particular organization stands within these paradigms. We also have introduced a change-management tool used to assess, plan, and monitor leadership effort and commitment to quality improvement and culture change activities, tracked as "momentum for change." This "momentum" is measured at eight discrete levels, from recognizing a performance gap to officially implementing changes intended to improve quality. PMID- 18406928 TI - Quality assurance needs for modern image-based radiotherapy: recommendations from 2007 interorganizational symposium on "quality assurance of radiation therapy: challenges of advanced technology". AB - This report summarizes the consensus findings and recommendations emerging from 2007 Symposium, "Quality Assurance of Radiation Therapy: Challenges of Advanced Technology." The Symposium was held in Dallas February 20-22, 2007. The 3-day program, which was sponsored jointly by the American Society for Therapeutic Radiology and Oncology (ASTRO), American Association of Physicists in Medicine (AAPM), and National Cancer Institute (NCI), included >40 invited speakers from the radiation oncology and industrial engineering/human factor communities and attracted nearly 350 attendees, mostly medical physicists. A summary of the major findings follows. The current process of developing consensus recommendations for prescriptive quality assurance (QA) tests remains valid for many of the devices and software systems used in modern radiotherapy (RT), although for some technologies, QA guidance is incomplete or out of date. The current approach to QA does not seem feasible for image-based planning, image-guided therapies, or computer-controlled therapy. In these areas, additional scientific investigation and innovative approaches are needed to manage risk and mitigate errors, including a better balance between mitigating the risk of catastrophic error and maintaining treatment quality, complimenting the current device-centered QA perspective by a more process-centered approach, and broadening community participation in QA guidance formulation and implementation. Industrial engineers and human factor experts can make significant contributions toward advancing a broader, more process-oriented, risk-based formulation of RT QA. Healthcare administrators need to appropriately increase personnel and ancillary equipment resources, as well as capital resources, when new advanced technology RT modalities are implemented. The pace of formalizing clinical physics training must rapidly increase to provide an adequately trained physics workforce for advanced technology RT. The specific recommendations of the Symposium included the following. First, the AAPM, in cooperation with other advisory bodies, should undertake a systematic program to update conventional QA guidance using available risk-assessment methods. Second, the AAPM advanced technology RT Task Groups should better balance clinical process vs. device operation aspects--encouraging greater levels of multidisciplinary participation such as industrial engineering consultants and use-risk assessment and process-flow techniques. Third, ASTRO should form a multidisciplinary subcommittee, consisting of physician, physicist, vendor, and industrial engineering representatives, to better address modern RT quality management and QA needs. Finally, government and private entities committed to improved healthcare quality and safety should support research directed toward addressing QA problems in image-guided therapies. PMID- 18406929 TI - Taxonometric applications in radiotherapy incident analysis. AB - Recent publications in both the scientific and the popular press have highlighted the risks to which patients expose themselves when entering a healthcare system. Patient safety issues are forcing us to, not only acknowledge that incidents do occur, but also actively develop the means for assessing and managing the risks of such incidents. To do this, we ideally need to know the probability of an incident's occurrence, the consequences or severity for the patient should it occur, and the basic causes of the incident. A structured approach to the description of failure modes is helpful in terms of communication, avoidance of ambiguity, and, ultimately, decision making for resource allocation. In this report, several classification schemes or taxonomies for use in risk assessment and management are discussed. In particular, a recently developed approach that reflects the activity domains through which the patient passes and that can be used as a basis for quantifying incident severity is described. The estimation of incident severity, which is based on the concept of the equivalent uniform dose, is presented in some detail. We conclude with a brief discussion on the use of a defined basic-causes table and how adding such a table to the reports of incidents can facilitate the allocation of resources. PMID- 18406930 TI - Taxonometric guidance for developing quality assurance. AB - A taxonomy is an ordered classification system. In error reduction analysis, an examination of the human failures that lead to an event often uses a taxonomy to classify the failures according to some aspect of their characteristics. These classifications provide insights into the forces that kept the persons involved from achieving their intended actions. They also can provide guidance for changing the situation to prevent failure in the future. For example, the corrective action would be different if a therapist pressed the wrong button because several buttons looked alike or they forgot which button to press. Different types of failures lead to particular types of remediation. Taxonomies can provide guidance in selecting between possible corrective actions. PMID- 18406931 TI - Variation and control of process behavior. AB - The purpose of this work was to highlight the importance of controlling process variability for successful quality assurance (QA). We describe the method of statistical process control for characterizing and controlling a process. Traditionally, QA has been performed by comparing some important measurement (e.g., linear accelerator output) against a corresponding specification. Although useful in determining the fitness of a particular measurement, this approach does not provide information about the underlying process behavior over time. A modern view of QA is to consider the time-ordered behavior of a process. Every process displays characteristic behaviors that are independent of the specifications imposed on it. The goal of modern QA is, not only to ensure that a process is on target, but that it is also operating with minimal variation. This is accomplished by way of a data-driven approach using process behavior charts. The development of process behavior charts, historically known as control charts, and process behavior (action) limits are described. The effect these concepts have on quality management is also discussed. PMID- 18406932 TI - Quality assurance of radiation therapy planning systems: current status and remaining challenges. AB - Computerized radiation therapy planning systems (RTPSs) are pivotal for treatment planning. The acceptance, commissioning, and quality control of RTPSs are uniquely complex and are described in the American Association of Physicists in Medicine Task Group Report 53 (1998) and International Atomic Energy Agency Technical Report Series No. 430 (2004). The International Atomic Energy Agency also developed a document and data package for use by vendors and purchasers to aid with acceptance testing of RTPSs. This document is based on International Electrotechnical Commission standard 62083 (2000) and describes both "type" tests to be performed in the factory and "site" tests to be performed in the clinic. The American Association of Physicists Task Group Report 67 described benchmark tests for the validation of dose calculation algorithms. Test data are being produced with the backing of the U.S. National Cancer Institute. However, significant challenges remain. Technology keeps evolving rapidly, thus requiring new quality assurance (QA) procedures. Intensity-modulated radiation therapy with its use of inverse optimization has added a new dimension to QA, because the results are not intuitively obvious. New technologies such as real-time ultrasound guidance for brachytherapy, TomoTherapy, and Cyberknife, require their own specialized RTPSs with unique QA requirements. On-line imaging allows for the generation of dose reconstructions using image warping techniques to determine the daily dose delivered to the patient. With increasing computer speeds, real time reoptimization of treatment plans will become a reality. Gating technologies will require four-dimensional dose calculations to determine the actual dose delivered to tissue voxels. With these rapidly changing technologies, it is essential that a strong QA culture is invoked in every institution implementing these procedures and that new protocols are developed as a part of the clinical implementation process. PMID- 18406933 TI - Developing quality assurance processes for image-guided adaptive radiation therapy. AB - Quality assurance has long been implemented in radiation treatment as systematic actions necessary to provide adequate confidence that the radiation oncology service will satisfy the given requirements for quality care. The existing reports from the American Association of Physicists in Medicine Task Groups 40 and 53 have provided highly detailed QA guidelines for conventional radiotherapy and treatment planning. However, advanced treatment processes recently developed with emerging high technology have introduced new QA requirements that have not been addressed previously in the conventional QA program. Therefore, it is necessary to expand the existing QA guidelines to also include new considerations. Image-guided adaptive radiation therapy (IGART) is a closed-loop treatment process that is designed to include the individual treatment information, such as patient-specific anatomic variation and delivered dose assessed during the therapy course in treatment evaluation and planning optimization. Clinical implementation of IGART requires high levels of automation in image acquisition, registration, segmentation, treatment dose construction, and adaptive planning optimization, which brings new challenges to the conventional QA program. In this article, clinical QA procedures for IGART are outlined. The discussion focuses on the dynamic or four-dimensional aspects of the IGART process, avoiding overlap with conventional QA guidelines. PMID- 18406934 TI - Quality assurance of serial 3D image registration, fusion, and segmentation. AB - Radiotherapy relies on images to plan, guide, and assess treatment. Image registration, fusion, and segmentation are integral to these processes; specifically for aiding anatomic delineation, assessing organ motion, and aligning targets with treatment beams in image-guided radiation therapy (IGRT). Future developments in image registration will also improve estimations of the actual dose delivered and quantitative assessment in patient follow-up exams. This article summarizes common and emerging technologies and reviews the role of image registration, fusion, and segmentation in radiotherapy processes. The current quality assurance practices are summarized, and implications for clinical procedures are discussed. PMID- 18406935 TI - Quality assurance of positron emission tomography/computed tomography for radiation therapy. AB - Recent advances in radiation delivery techniques, such as intensity-modulated radiation therapy, provide unprecedented ability to exquisitely control three dimensional dose distribution. Development of on-board imaging and other image guidance methods significantly improved our ability to better target a radiation beam to the tumor volume. However, in reality, accurate definition of the location and boundary of the tumor target is still problematic. Biologic and physiologic imaging promises to solve the problem in a fundamental way and has a more and more important role in patient staging, treatment planning, and therapeutic assessment in radiation therapy clinics. The last decade witnessed a dramatic increase in the use of positron emission tomography and computed tomography in radiotherapy practice. To ensure safe and effective use of nuclide imaging, a rigorous quality assurance (QA) protocol of the imaging tools and integration of the imaging data must be in place. The application of nuclide imaging in radiation oncology occurs at different levels of sophistication. Quantitative use of the imaging data in treatment planning through image registration and standardized uptake value calculation is often involved. Thus, QA should not be limited to the performance of the scanner, but should also include the process of implementing image data in treatment planning, such as data transfer, image registration, and quantitation of data for delineation of tumors and sensitive structures. This presentation discusses various aspects of nuclide imaging as applied to radiotherapy and describes the QA procedures necessary for the success of biologic image-guided radiation therapy. PMID- 18406936 TI - Newer imaging modalities to assist with target localization in the radiation treatment of prostate cancer and possible lymph node metastases. AB - Precise localization of prostate cancer and the drainage lymph nodes is mandatory to define an accurate clinical target volume for conformal radiotherapy. Better target definition and delineation on a daily basis is surely important in quality assurance for fractionated radiation therapy. This article reviews the evidence for major emerging techniques that show promise in better identifying the clinical target volume. Partial prostate boost by brachytherapy, intensity modulated radiation therapy, or protons has become possible not only with standard imaging techniques but also with the availability of metabolic images obtained by magnetic resonance spectroscopy. Even though fluorine-18 fluorodeoxyglucose ((18)F-FDG) positron emission tomography has not been found to be useful, novel radiolabeled tracers may eventually prove of value in the diagnosis and treatment planning of prostate cancer. For the metastatic lymph nodes, lymphotropic nanoparticle-enhanced magnetic resonance imaging using ultra small superparamagnetic iron oxide particles has greater accuracy as compared with conventional techniques and has been instrumental in delineating the lymphatic drainage of the prostate gland. These novel investigational techniques could further help in optimizing conformal radiotherapy for patients with prostate cancer. The concepts of biologic target volume, real target volume, and multidimensional conformal radiotherapy are being explored. PMID- 18406937 TI - Quality assurance for kilo- and megavoltage in-room imaging and localization for off- and online setup error correction. AB - In-room radiography is not a new concept for image-guided radiation therapy. Rapid advances in technology, however, have made this positioning method convenient, and thus radiograph-based positioning has propagated widely. The paradigms for quality assurance of radiograph-based positioning include imager performance, systems integration, infrastructure, procedure documentation and testing, and support for positioning strategy implementation. PMID- 18406938 TI - Quality assurance of ultrasound imaging systems for target localization and online setup corrections. AB - We describe quality assurance paradigms for ultrasound imaging systems for target localization (UISTL). To determine the absolute localization accuracy of a UISTL, an absolute coordinate system can be established in the treatment room and spherical targets at various depths can be localized. To test the ability of such a system to determine the magnitude of internal organ motion, a phantom that mimics the human male pelvic anatomy can be used to simulate different organ motion ranges. To assess the interuser variability of ultrasound (US) guidance, different experienced users can independently determine the daily organ shifts for the same patients for a number of consecutive fractions. The average accuracy for a UISTL for the localization of spherical targets at various depths has been found to be 0.57 +/- 0.47 mm in each spatial dimension for various focal depths. For the phantom organ motion test it was found that the true organ motion could be determined to within 1.0 mm along each axis. The variability between different experienced users who localized the same 5 patients for five consecutive fractions was small in comparison to the indicated shifts. In addition to the quality assurance tests that address the ability of a UISTL to accurately localize a target, a thorough quality assurance program should also incorporate the following two aspects to ensure consistent and accurate localization in daily clinical use: (1) adequate training and performance monitoring of users of the US target localization system, and (2) prescreening of patients who may not be good candidates for US localization. PMID- 18406939 TI - Quality assurance for the geometric accuracy of cone-beam CT guidance in radiation therapy. AB - The introduction of volumetric X-ray image-guided radiotherapy systems allows improved management of geometric variations in patient setup and internal organ motion. As these systems become a routine clinical modality, we propose a daily quality assurance (QA) program for cone-beam computed tomography (CBCT) integrated with a linear accelerator. The image-guided system used in this work combines a linear accelerator with conventional X-ray tube and an amorphous silicon flat-panel detector mounted orthogonally from the accelerator central beam axis. This article focuses on daily QA protocols germane to geometric accuracy of the CBCT systems and proposes tolerance levels on the basis of more than 3 years of experience with seven CBCT systems used in our clinic. Monthly geometric calibration tests demonstrate the long-term stability of the flex movements, which are reproducible within +/-0.5 mm (95% confidence interval). The daily QA procedure demonstrates that, for rigid phantoms, the accuracy of the image-guided process can be within 1 mm on average, with a 99% confidence interval of +/-2 mm. PMID- 18406940 TI - Quality assurance of onboard megavoltage computed tomography imaging and target localization systems for on- and off-line image-guided radiotherapy. AB - We reviewed the quality assurance procedures that have been used to test fan- and cone-beam megavoltage-based in-room imaging systems. Phantom-based tests have been used to establish the geometric accuracy and precision of megavoltage-based systems. However, the clinical implementation of any system is accompanied by challenges that are best tested in a clinical setting using clinical images. To objectively judge and monitor image quality, a set of standard tests and phantoms can be used. The image noise and spatial and contrast resolution have been assessed using standard computed tomography phantoms. The dose to the patient resulting from the imaging procedure can be determined using calculations or measurements. The off-line use of patient images is of interest for the evaluation of dosimetric changes throughout the treatment course. The accuracy of the dosimetric calculations based on the megavoltage images has been tested for the fan- and cone-beam systems. Some of the described tests are typically performed before the clinical implementation of the imaging system; others are suited to monitor the system's performances. PMID- 18406941 TI - Quality assurance issues in conducting multi-institutional advanced technology clinical trials. AB - The National Cancer Institute-sponsored Advanced Technology Quality Assurance (QA) Consortium, which consisted of the Image-Guided Therapy QA Center, Radiation Therapy Oncology Group, Radiological Physics Center, Quality Assurance Review Center, and Resource Center for Emerging Technologies, has pioneered the development of an infrastructure and QA method for advanced technology clinical trials that requires volumetric digital data submission of a protocol patient's treatment plan and verification data. In particular, the Image-Guided Therapy QA Center has nearly 15 years experience in facilitating QA review for Radiation Therapy Oncology Group advanced technology clinical trials. This QA process includes (1) a data integrity review for completeness of protocol required elements, the format of data, and possible data corruption, and recalculation of dose-volume histograms; (2) a review of compliance with target volume and organ at-risk contours by study chairs; and (3) a review of dose prescription and dose heterogeneity compliance by the Radiation Therapy Oncology Group Headquarters Dosimetry Group or the Radiological Physics Center dosimetrists (for brachytherapy protocols). This report reviews the lessons learned and the QA challenges presented by the use of advanced treatment modalities in clinical trials requiring volumetric digital data submission. PMID- 18406942 TI - Challenges in credentialing institutions and participants in advanced technology multi-institutional clinical trials. AB - The Radiological Physics Center (RPC) has functioned continuously for 38 years to assure the National Cancer Institute and the cooperative groups that institutions participating in multi-institutional trials can be expected to deliver radiation treatments that are clinically comparable to those delivered by other institutions in the cooperative groups. To accomplish this, the RPC monitors the machine output, the dosimetry data used by the institutions, the calculation algorithms used for treatment planning, and the institutions' quality control procedures. The methods of monitoring include on-site dosimetry review by an RPC physicist and a variety of remote audit tools. The introduction of advanced technology clinical trials has prompted several study groups to require participating institutions and personnel to become credentialed, to ensure their familiarity and capability with techniques such as three-dimensional conformal radiotherapy, intensity-modulated radiotherapy, stereotactic body radiotherapy, and brachytherapy. The RPC conducts a variety of credentialing activities, beginning with questionnaires to evaluate an institution's understanding of the protocol and their capabilities. Treatment-planning benchmarks are used to allow the institution to demonstrate their planning ability and to facilitate a review of the accuracy of treatment-planning systems under relevant conditions. The RPC also provides mailable anthropomorphic phantoms to verify tumor dose delivery for special treatment techniques. While conducting these reviews, the RPC has amassed a large amount of data describing the dosimetry at participating institutions. Representative data from the monitoring programs are discussed, and examples are presented of specific instances in which the RPC contributed to the discovery and resolution of dosimetry errors. PMID- 18406943 TI - Processes for quality improvements in radiation oncology clinical trials. AB - Quality assurance in radiotherapy (RT) has been an integral aspect of cooperative group clinical trials since 1970. In early clinical trials, data acquisition was nonuniform and inconsistent and computational models for radiation dose calculation varied significantly. Process improvements developed for data acquisition, credentialing, and data management have provided the necessary infrastructure for uniform data. With continued improvement in the technology and delivery of RT, evaluation processes for target definition, RT planning, and execution undergo constant review. As we move to multimodality image-based definitions of target volumes for protocols, future clinical trials will require near real-time image analysis and feedback to field investigators. The ability of quality assurance centers to meet these real-time challenges with robust electronic interaction platforms for imaging acquisition, review, archiving, and quantitative review of volumetric RT plans will be the primary challenge for future successful clinical trials. PMID- 18406944 TI - International perspectives on quality assurance and new techniques in radiation medicine: outcomes of an IAEA conference. AB - The International Atomic Energy Agency organized an international conference called, "Quality Assurance and New Techniques in Radiation Medicine" (QANTRM). It dealt with quality assurance (QA) in all aspects of radiation medicine (diagnostic radiology, nuclear medicine, and radiotherapy) at the international level. Participants discussed QA issues pertaining to the implementation of new technologies and the need for education and staff training. The advantage of developing a comprehensive and harmonized approach to QA covering both the technical and the managerial issues was emphasized to ensure the optimization of benefits to patient safety and effectiveness. The necessary coupling between medical radiation imaging and radiotherapy was stressed, particularly for advanced technologies. However, the need for a more systematic approach to the adoption of advanced technologies was underscored by a report on failures in intensity-modulated radiotherapy dosimetry auditing tests in the United States, which could imply inadequate implementation of QA for these new technologies. A plenary session addressed the socioeconomic impact of introducing advanced technologies in resource-limited settings. How shall the dual gaps, one in access to basic medical services and the other in access to high-quality modern technology, be addressed? PMID- 18406945 TI - IMRT delivery performance with a varian multileaf collimator. AB - The use of a multileaf collimator (MLC) for intensity-modulated radiotherapy poses unique dosimetric issues. The nature of intensity-modulated radiotherapy dosimetry, centered on leaf position accuracy, is common to all MLCs. However, the mechanical and software designs of MLCs from the different manufacturers distinguish them. This report focused on the Varian Millennium 120, although the concepts are applicable to the earlier Varian Mark series as well. The factors that affect dose delivery in clinical fields (i.e., mechanical tolerances, motor fatigue, and latency effects) have been quantified. Moreover, inadequate modeling of the MLC in the planning system can be perceived as erratic performance. Individually, some problems have been shown to be insignificant; others are correctable using software. If these problems are rectified or at least understood by the physicist, quality assurance can be simplified. PMID- 18406946 TI - Multileaf collimator characteristics and reliability requirements for IMRT Elekta system. AB - Understanding the characteristics of a multileaf collimator (MLC) system, modeling MLC in a treatment planning system, and maintaining the mechanical accuracy of the linear accelerator gantry head system are important factors in the safe implementation of an intensity-modulated radiotherapy program. We review the characteristics of an Elekta MLC system, discuss the necessary MLC modeling parameters for a treatment planning system, and provide a novel method to establish an MLC leaf position quality assurance program. To perform quality assurance on 40 pairs of individual MLC leaves is a time-consuming and difficult task. In this report, an effective routine MLC quality assurance method based on the field edge of a backup jaw as referenced in conjunction with a diode array as a radiation detector system is discussed. The sensitivity of this test for determining the relative leaf positions was observed to be better than 0.1 mm. The Elekta MLC leaf position accuracy measured with this system has been better than 0.3 mm. PMID- 18406947 TI - Siemens multileaf collimator characterization and quality assurance approaches for intensity-modulated radiotherapy. AB - Application of the multileaf collimator (MLC) has evolved from replacing blocks to create treatment fields to creating photon fluence modulation for intensity modulated radiotherapy (IMRT). Multileaf collimator system performance requirements are far more stringent for such applications and will require increased performance for future applications, such as motion tracking. This article reviews Siemens MLC systems, including a technical description and dosimetric characteristics of 56-, 82-, and 160-leaf designs. Routine quality assurance of MLC for IMRT necessitates frequent and critical assessment of MLC leaf position calibration errors that can present in many different ways (e.g., accuracy, reproducibility, longevity, hysteresis, and collimator/gantry angle dependencies). Several techniques for measuring these errors are presented, along with qualitative and quantitative techniques for analyzing results. In particular, increased accuracy of leaf position measurement at variable gantry angles is enabled by spatial transformations to electronic portal imaging device position quantified by calibration protocols introduced with megavoltage cone beam. Measured values of X-ray transmission (intra-leaf, inter-leaf, and through abutting leaf pairs) and penumbra (leaf end, leaf tongue, leaf groove) are presented with an evaluation of their characterization by a treatment-planning system. The dosimetric impact of planning system model inadequacies is demonstrated for collimator scatter, dose profile values within 30 mm of the field edge, and the resultant effect demonstrated on clinical cases. Finally, a description of automated quality assurance delivery, analysis, and calibration protocols applicable for the specific vendor's system is provided. PMID- 18406948 TI - QA issues for computer-controlled treatment delivery: this is not your old R/V system any more! AB - State-of-the-art radiotherapy treatment delivery has changed dramatically during the past decade, moving from manual individual field setup and treatment to automated computer-controlled delivery of complex treatments, including intensity modulated radiotherapy and other similarly complex delivery strategies. However, the quality assurance methods typically used to ensure treatment is performed precisely and correctly have not evolved in a similarly dramatic way. This paper reviews the old manual treatment process and use of record-and-verify systems, and describes differences with modern computer-controlled treatment delivery. The process and technology used for computer-controlled treatment delivery are analyzed in terms of potential (and actual) problems, as well as relevant published guidance on quality assurance. The potential for improved quality assurance for computer-controlled delivery is discussed. PMID- 18406949 TI - Arthroscopic versus open dorsal ganglion excision: a prospective, randomized comparison of rates of recurrence and of residual pain. AB - PURPOSE: The purpose of this study was to compare the postoperative rates of ganglion recurrence between arthroscopic and open techniques of dorsal ganglion (DG) excision. METHODS: A total of 72 patients had either arthroscopic or open excision of a primary, simple DG by 1 of 2 senior hand surgeons. Three prospective postoperative assessments were performed. The first examination was performed at 5 to 7 days, the second at 4 to 8 weeks, and the third assessment was performed at a minimum of 1 year after surgery. Percentages of ganglion recurrence at the second and third assessments were recorded. RESULTS: Forty-one patients had arthroscopic excision, and 31 patients had open excision. Baseline patient age, gender, and surgical side were similar between the 2 groups. Recurrence of the DG at the second postoperative assessment was 1 of 41 patients in the arthroscopic group and none in the open excision group, and, after a minimum of 12 months after excision, recurrence was 3 of 28 in the arthroscopic group and 2 of 23 in the open group. CONCLUSIONS: This study compares the rates of ganglion recurrence between arthroscopic and open DG excision. Our results demonstrate that at 12 months follow-up, the rates of recurrence with arthroscopic DG excision are comparable with and not superior to those of open excision. Our results suggest that additional long-term comparative studies are needed to accurately differentiate the efficacy of open and arthroscopic techniques. PMID- 18406950 TI - Arthroscopic assessment of intra-articular distal radius fractures after open reduction and internal fixation from a volar approach. AB - PURPOSE: The volar approach with locked plating is a common treatment for intra articular distal radius fractures. The purpose of this study was to arthroscopically assess the articular surface after internal fixation through the volar approach as a means to evaluate the ability of an extra-articular reduction to anatomically restore the joint surface. METHODS: Sixteen patients with intra articular distal radius fractures were prospectively enrolled. A volar approach and internal fixation using a locked volar plate was performed. Using a visual analog scale (VAS), the fracture reduction was clinically graded on the quality of reduction of the visible metaphyseal fracture lines, fluoroscopically graded, and arthroscopic graded. Maximum step and gap deformity were recorded from arthroscopy and plain radiograph. RESULTS: The mean VAS score for the fracture reduction based on extra-articular fracture lines was 7.4. The mean VAS score for the fluoroscopic reduction was 8.2. The mean VAS score for the arthroscopic reduction was 6.4. The arthroscopic VAS score was significantly lower than the VAS score for fluoroscopy but was not significantly different than the VAS score for metaphyseal reduction. Mean arthroscopic measurement of maximum step and gap deformity were 1 mm and 2 mm, respectively. Mean postoperative radiographic maximum step and gap deformity were both less than 1 mm. The arthroscopic step and gap deformities were significantly greater than the radiographic deformities. CONCLUSIONS: A volar approach, indirect reduction, and locked plate fixation is a useful technique in restoring articular congruity after distal radius fracture. The number of fracture lines and presence of step and gap deformity can be adequately assessed using clinical and fluoroscopic assessment. However, the magnitude of step and gap deformity may be underestimated. PMID- 18406951 TI - Functional disability of the wrist: direct correlation with decreased wrist motion. AB - PURPOSE: Motion-limiting wrist procedures have demonstrated both efficacy and reproducibility as a treatment for alleviating painful wrist conditions, but the reduction of pain achieved is necessarily accompanied by a marked loss of wrist motion. However, no study demonstrates a functional difference between variable degrees of wrist motion. The hypothesis of this study is that wrist motion is directly correlated with functional ability. METHODS: Using a prospective, randomized, crossover design, we randomized 42 men and women above 45 years of age with normal wrists to wear both a partially restricted and a highly restricted splint for 24 hours each. Serving as their own controls, objective and subjective outcome measurements were taken at baseline and after each of the 2 splinting periods. The specific measurements included range of motion parameters (flexion/extension, radioulnar deviation, supination/pronation, circumduction), subjective surveys (Disabilities of the Arm, Shoulder, and Hand; Patient Rated Wrist Evaluation; and Modern Activity Subjective Survey), and an objective timed test (Modern Activity Timed Test; MATT). RESULTS: The simulated flexion/extension arc of motion for baseline, partially restricted, and highly restricted conditions were 138 degrees , 58 degrees , and 20 degrees , respectively. Median Disabilities of the Arm, Shoulder, and Hand scores for the 3 conditions significantly increased from 1.8 to 14.4 and 20.8 (p <.01), respectively. In addition, Patient Rated Wrist Evaluation results increased from 2.5 to 16.4 and 27.1, and the Modern Activity Subjective Survey results increased from 0.3 to 9.1 and 14.9 across restricted motion conditions. Modern Activity Timed Test results matched the perceived difficulty reflected by the subjective surveys, demonstrating significant increases in performance time for 6 of the 8 individual MATT items as well as significant differences for the sum of all tasks (MATT score) between the highly and partially restricted conditions. CONCLUSIONS: Use of low-profile splinting to restrict wrist motion demonstrates a direct correlation between restriction of wrist motion and functional disability. PMID- 18406952 TI - Treatment of advanced-stage Kienbock's disease with proximal row carpectomy: an average 15-year follow-up. AB - PURPOSE: The purpose of this study was to evaluate the long-term clinical results of a proximal row carpectomy with an average 15-year follow-up for the primary treatment of advanced-stage (Lichtman IIIA and IIIB) Kienbock's disease. This study is a follow-up to a paper by the senior author in a similar cohort of patients where the clinical results of a proximal row carpectomy were evaluated for the treatment of advanced-stage Kienbock's disease at an average 3-year follow-up. METHODS: Seventeen patients with a minimum of 10 years of follow-up were identified who met criteria for inclusion. Thirteen of these patients were located and agreed to participate. Patients were seen, examined, and queried regarding their wrists. Range of motion, grip strength, and subjective patient satisfaction were all obtained and quantified using a clinical outcomes scale. RESULTS: Twelve of 13 patients demonstrated excellent or good results based on the clinical outcomes scale used (5 excellent, 7 good, 1 fair, and none poor). Total arc of motion averaged 73% of the uninvolved side. Grip strength averaged 92% of the uninvolved side. Compared with preoperative values, range of motion improved an average of 16% and grip strength improved an average of 129%, an overall average improvement of 12 degrees and 18 kg, respectively. At the most recent follow-up, all patients remained employed. Seven patients held manual labor positions, 2 were nurses, and 4 were employed in sedentary vocations. All patients demonstrated some degree of degenerative changes, usually localized to the radiocapitate articulation in the lunate fossa. Clinical results did not correlate with radiographic degeneration. CONCLUSIONS: This study demonstrates proximal row carpectomy to be a reliable motion-preserving procedure with good clinical results maintained out to an average of 15 years postoperatively. PMID- 18406953 TI - Simulated radioscapholunate fusion alters carpal kinematics while preserving dart thrower's motion. AB - PURPOSE: Midcarpal degeneration is well documented after radioscapholunate fusion. This study tested the hypothesis that radioscapholunate fusion alters the kinematic behavior of the remaining lunotriquetral and midcarpal joints, with specific focus on the dart-thrower's motion. METHODS: Simulated radioscapholunate fusions were performed on 6 cadaveric wrists in an anatomically neutral posture. Two 0.060-in. carbon fiber pins were placed from proximal to distal across the radiolunate and radioscaphoid joints, respectively. The wrists were passively positioned in a custom jig toward a full range of motion along the orthogonal axes as well as oblique motions, with additional intermediate positions along the dart-thrower's path. Using a computed tomography-based markerless bone registration technique, each carpal bone's three-dimensional rotation was defined as a function of wrist flexion/extension from the pinned neutral position. Kinematic data was analyzed against data collected on the same wrist prior to fixation using hierarchical linear regression analysis and paired Student's t tests. RESULTS: After simulated fusion, wrist motion was restricted to an average flexion-extension arc of 48 degrees , reduced from 77 degrees , and radial-ulnar deviation arc of 19 degrees , reduced from 33 degrees . The remaining motion was maximally preserved along the dart-thrower's path from radial-extension toward ulnar-flexion. The simulated fusion significantly increased rotation through the scaphotrapezial joint, scaphocapitate joint, triquetrohamate joint, and lunotriquetral joint. For example, in the pinned wrist, the rotation of the hamate relative to the triquetrum increased 85%. Therefore, during every 10 degrees of total wrist motion, the hamate rotated an average of nearly 8 degrees relative to the triquetrum after pinning versus 4 degrees in the normal state. CONCLUSIONS: Simulated radioscapholunate fusion altered midcarpal and lunotriquetral kinematics. The increased rotations across these remaining joints provide one potential explanation for midcarpal degeneration after radioscapholunate fusion. Additionally, this fusion model confirms the dart thrower's hypothesis, as wrist motion after simulated radioscapholunate fusion was primarily preserved from radial-extension toward ulnar-flexion. PMID- 18406954 TI - Clinical outcome and histologic findings of costal osteochondral grafts for cartilage defects in finger joints. AB - PURPOSE: For the purpose of achieving anatomical reduction as precisely as possible, we performed osteochondral grafting from the costo-osteochondral junction in 16 patients (17 joints) with posttraumatic articular cartilage injury or avascular necrosis in finger joints. The purpose of this study was to review our series of costal osteochondral grafts in order to determine the practicality, effectiveness, and functionality of this grafting technique in a clinical setting. METHODS: Patients were followed for at least 18 months postoperatively (18-57 months; average, 28 months). The injured joints included 3 metacarpophalangeal, 9 proximal interphalangeal, 3 distal interphalangeal, and 2 thumb interphalangeal joints. The defect accounted for 50% to 100% of the articular surface (average, 63%). RESULTS: The average time until bone union of the graft was 58 days. The mean arc of motion was 13 degrees before surgery versus 58 degrees after surgery, with a mean increase of 45 degrees . In 7 patients (8 joints), an extremely small portion (approximately 1 x 1 mm in size and thinner than 0.1 mm) of the implanted cartilage was obtained via biopsy using a scalpel with the consent of the patient at the time of screw removal and was used to prepare histologic specimens, which revealed scattered chondrocytes within the matrix without differences from normal hyaline cartilage in any. The chondrocytes in the grafts appeared viable, and the reconstruction of the joint surface could be confirmed histologically. CONCLUSIONS: Osteochondral grafting from the costo-osteochondral junction achieves excellent reconstruction of the injured joint without affecting other joints. This technique is particularly beneficial in cases where it is difficult to obtain allograft donors, as is often the case in Japan. Despite these encouraging findings in this small series, we believe that it is necessary to conduct further studies of this method over a longer period. PMID- 18406955 TI - A prospective randomized controlled trial of injection of dexamethasone versus triamcinolone for idiopathic trigger finger. AB - PURPOSE: This study was designed to test the null hypothesis that there is no difference in resolution of triggering 3 months after injection with either a soluble (dexamethasone) or insoluble (triamcinolone) corticosteroid for idiopathic trigger finger. METHODS: Eighty-four patients were enrolled in a prospective randomized controlled trial comparing dexamethasone and triamcinolone injection for idiopathic trigger finger. Sixty-seven patients completed the 6 week follow-up (35 triamcinolone arm, 32 dexamethasone arm), and 72 patients completed the 3-month follow-up (41 triamcinolone arm, 31 dexamethasone arm). Outcome measures included the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire, trigger finger grading according to Quinnell, and satisfaction on a visual analog scale. To preserve autonomy, patients were permitted additional injections and operative treatment at any time. Twenty-five patients requested a second injection (10 triamcinolone arm, 15 dexamethasone arm), and 21 elected operative treatment (10 triamcinolone arm, 11 dexamethasone arm) during the study period. The analysis was according to intention to treat principles. RESULTS: Six weeks after injection, absence of triggering was documented in 22 of 35 patients in the triamcinolone cohort and in 12 of 32 patients in the dexamethasone cohort. The rates 3 months after injection were 27 of 41 in the triamcinolone cohort and 22 of 31 in the dexamethasone cohort. The triamcinolone cohort had significantly better satisfaction and Quinnell grades than did the dexamethasone cohort at the 6-week follow-up but not at the 3-month follow-up. There were no significant differences between Disabilities of the Arm, Shoulder, and Hand scores at the 6 week follow-up and the 3-month follow-up. After the close of the study, there were 8 recurrences among patients with documented absence of triggering in the triamcinolone cohort and 1 in the dexamethasone cohort. CONCLUSIONS: Although there were no differences 3 months after injection, our data suggest that triamcinolone may have a more rapid but ultimately less durable effect on idiopathic trigger finger than does dexamethasone. PMID- 18406957 TI - The quality and strength of evidence for etiology: example of carpal tunnel syndrome. AB - PURPOSE: The purpose of this investigation was to evaluate the quality and strength of scientific evidence supporting an etiologic relationship between a disease and a proposed risk factor using a scoring system based on the Bradford Hill criteria for causal association. METHODS: A quantitative score based on the Bradford Hill criteria (qBHs) was used to evaluate 117 articles presenting original data regarding the etiology of carpal tunnel syndrome: 33 (28%) that evaluated biological (structural or genetic) risk factors, 51 (44%) that evaluated occupational (environment or activity-related) risk factors, and 33 (28%) that evaluated both types of risk factors. RESULTS: The quantitative Bradford Hill scores of 2 independent observers showed very good agreement, supporting the reliability of the instrument. The average qBHs was 12.2 points (moderate association) among biological risk factors compared with 5.2 points (poor association) for occupational risk factors. The highest average qBHs was observed for genetic factors (14.2), race (11.7), and anthropometric measures of the wrist (11.3 points) with all studies finding a moderate causal association. The highest average qBHs among occupational risk factors was observed for activities requiring repetitive hand use (6.5 points among the 30 of 45 articles that reported a causal association), substantial exposure to vibration (6.3 points; 14 of 20 articles), and type of occupation (5.6 points; 38 of 53 articles), with the findings being much less consistent. CONCLUSIONS: According to a quantitative analysis of published scientific evidence, the etiology of carpal tunnel syndrome is largely structural, genetic, and biological, with environmental and occupational factors such as repetitive hand use playing a minor and more debatable role. Speculative causal theories should be analyzed through a rigorous approach prior to wide adoption. PMID- 18406958 TI - An implanted upper-extremity neuroprosthesis using myoelectric control. AB - PURPOSE: The purpose of this study was to evaluate the potential of a second generation implantable neuroprosthesis that provides improved control of hand grasp and elbow extension for individuals with cervical level spinal cord injury. The key feature of this system is that users control their stimulated function through electromyographic (EMG) signals. METHODS: The second-generation neuroprosthesis consists of 12 stimulating electrodes, 2 EMG signal recording electrodes, an implanted stimulator-telemeter device, an external control unit, and a transmit/receive coil. The system was implanted in a single surgical procedure. Functional outcomes for each subject were evaluated in the domains of body functions and structures, activity performance, and societal participation. RESULTS: Three individuals with C5/C6 spinal cord injury received system implantation with subsequent prospective evaluation for a minimum of 2 years. All 3 subjects demonstrated that EMG signals can be recorded from voluntary muscles in the presence of electrical stimulation of nearby muscles. Significantly increased pinch force and grasp function was achieved for each subject. Functional evaluation demonstrated improvement in at least 5 activities of daily living using the Activities of Daily Living Abilities Test. Each subject was able to use the device at home. There were no system failures. Two of 6 EMG electrodes required surgical revision because of suboptimal location of the recording electrodes. CONCLUSIONS: These results indicate that a neuroprosthesis with implanted myoelectric control is an effective method for restoring hand function in midcervical level spinal cord injury. PMID- 18406959 TI - Chronic hand ischemia treated with radial artery balloon angioplasty: case report. AB - Although balloon angioplasty in heart and lower extremity vessels has been extensively studied and reported, little information exists regarding its use for digital ischemia in the hand. We report a case of successful balloon angioplasty of the distal radial artery to reverse present and prevent further digital tip cyanosis and necrosis. PMID- 18406960 TI - Complex dorsal metacarpophalangeal joint dislocation caused by interosseous tendon entrapment: case report. AB - We report a patient with open complex, dorsal metacarpophalangeal joint dislocation of the index finger with interposition of the first dorsal interosseous tendon and the transverse ligament of the palmar aponeurosis. To our knowledge, entrapment of the first dorsal interosseous tendon has not been reported to be the cause of irreducible dislocation of the metacarpophalangeal joint. PMID- 18406961 TI - Grip strength in healthy caucasian adults: reference values. AB - PURPOSE: The aim of this study was to update reference data of handgrip strength for healthy adults of both genders spanning a wide age range and to analyze possible factors of influence. METHODS: Intraindividual and interindividual variations of grip strength and their relation to several anthropometric factors were analyzed in a standardized manner for 769 healthy adults (women, n = 403; men, n = 366) aged between 20 years and 95 years. Measurements were done in neutral position of arm, forearm, and wrist on setting II of a Baseline digital hydraulic dynamometer (NexGen Ergonomics Inc. Quebec, Canada). RESULTS: Mean strength was about 41% less in women (right 29 kg; left 27 kg) than in men (right 49 kg; left 47 kg) resulting in a ratio of left to right hand slightly above .95 in both genders. During the course of life, hand strength develops comparably in both genders peaking at 35 years of age and decreasing continuously further on. Anthropometric variables such as forearm circumference and length, hand size, or body mass showed a positive correlation with grip strength. Body mass index, type of work, and hand dominance showed only a partial positive correlation or no correlation with grip strength. Gender and age, followed by parameters representing body length and obesity, were observed to have the highest predictive value for handgrip strength and were therefore entered into the generation of prediction equations. CONCLUSIONS: We recommend side adjustment of measured values for intraindividual comparison and inclusion of information regarding anthropometric characteristics, as well as using gender- and age adjusted reference values, whereas hand dominance can be neglected. The regression equations we generated might prove to be useful for clinicians or for those who use normative values within software to provide more accurate predictions of strength scores for specific applications. PMID- 18406962 TI - Fundamental principles of writing a successful grant proposal. AB - It is important for the field of hand surgery to develop a new generation of surgeon-scientists who can produce high-impact studies to raise the profile of this specialty. To this end, organizations such as the American Society for Surgery of the Hand have initiated programs to promote multicenter clinical research that can be competitive for fiscal support from the National Institutes of Health and other funding agencies. Crafting a well-structured grant proposal is critical to securing adequate funding to investigate the many clinical and basic science questions in hand surgery. In this article, we present the key elements of a successful grant proposal to help potential applicants to navigate the complex pathways in the grant application process. PMID- 18406963 TI - Management of intra-articular metacarpal base fractures of the second through fifth metacarpals. AB - Intra-articular fractures of the second through fifth metacarpal bases are uncommon injuries but can result in serious morbidity if improperly managed. These injuries usually occur because of forced flexion of the wrist with simultaneous extension of the arm, as occurs with a punch or a fall. As there are few large series of reports for these injuries, there is no consensus in the current literature on the most appropriate treatment for them. Whereas some authors have reported successful results after closed reduction, many recommend open reduction with internal fixation to ensure the integrity of the tendinous insertions of the extensor carpi radialis longus, extensor carpi radialis brevis, and extensor carpi ulnaris. This article reviews the case reports and case series extant in the literature concerning intra-articular fractures of the bases of the second through fifth metacarpals, and it provides important diagnostic and management considerations for these injuries. PMID- 18406964 TI - Physeal transfers for skeletal reconstruction. AB - In this article, we present the common methods of vascularized and nonvascularized physeal transfers with an emphasis on vascularized fibula physeal transfers. Physeal transfers are important in skeletally immature patients in whom continued growth of the limb is desired. Physeal transfers now represent the state of the art method for reconstruction of pediatric long bones for reasons including tumor resection, congenital deformities and trauma. This form of reconstruction can produce functional and durable results. PMID- 18406965 TI - Noninvasive evaluation of upper-extremity vascular perfusion. AB - Evaluation of vascular disorders of the upper extremity requires an anatomic and functional approach. The combination of a good history and physical examination, laboratory testing, and specialized vascular laboratory studies will help detect any underlying collagen vascular disease or coagulopathy and provide physiologic specific and patient-oriented management. This paper reviews the currently available noninvasive modalities for the evaluation of upper-extremity perfusion. PMID- 18406966 TI - Radiation injury and the hand surgeon. AB - The human hand has been affected by ionizing radiation accidents than any other organ. Hand surgeons should understand the pathophysiology and appropriate management of various types of radiation injury. This article outlines the history and epidemiology of ionizing radiation injury to the hand, basic aspects of radiobiology, and principles of management for injury resulting from fluoroscopy, nuclear accidents or weapons, and other sources. PMID- 18406967 TI - A technique of direct tracing for recording digital range of motion. AB - This report describes a technique for recording range of motion of digital joints. The technique involves tracing digital outlines with an angled marker on a sheet of paper attached to a special interray pad. A complete dorsal silhouette of the entire ray can be obtained by mounting the pad and the paper astride the palm parallel to the ray of the digit being evaluated. Fabrication and use of the equipment is described. This silhouetting system could be an alternative to wire tracing in visualizing tendencies of dynamics of digital range of motion. PMID- 18406968 TI - Endoscopic decompression of the ulnar nerve at the elbow. PMID- 18406972 TI - Infectious diseases in emergency medicine. Foreword. PMID- 18406970 TI - Palmar bypass for digital ischemia. PMID- 18406973 TI - Infectious diseases in emergency medicine. Preface. PMID- 18406974 TI - Timing of antibiotics for acute, severe infections. AB - The importance of antibiotic timing is a common clinical question encountered in emergency medicine practice for patients who have severe infections. Various studies in the medical literature have reported associations between early antibiotic timing and improved survival for meningitis, pneumonia, and septic shock. Understanding the evidence behind antibiotic timing and survival is vital to emergency physicians, because they must balance the potential benefits of early antibiotic administration and the potential for antibiotic overuse and misuse. The measurement of antibiotic timing in pneumonia has been shown to be associated with antibiotic misuse in emergency departments. Quality organizations should study carefully the intended and unintended consequences of measuring and reporting antibiotic timing to make policy decisions on current and future performance measures in this area. PMID- 18406975 TI - Rapidly fatal infections. AB - Emergency physicians are trained to separate "sick" from "not sick" patients during their training. Nevertheless, every emergency physician will face situations in which early intervention is critical to their patient's outcome. Infectious diseases are responsible for many of these potentially poor outcomes. This article discusses early identification and treatment for several rapidly fatal infections, including two newly identified travel-related illnesses. PMID- 18406976 TI - Acute bacterial meningitis. AB - Despite advances in medical care including antibiotics and vaccines, meningitis still has a high morbidity and mortality rate, especially in certain high-risk patients. Early diagnosis with the administration of appropriate antibiotics remains the key element of management. This article highlights methods of diagnosis, differential diagnoses, treatment options, and complications of treating bacterial meningitis. PMID- 18406977 TI - Evaluation and management of geriatric infections in the emergency department. AB - A significant proportion of elderly emergency department visits are for acute infections. The elderly are more likely than younger patients to have nonspecific signs and symptoms of infection, more severe disease, and resistant microorganisms. As a result, emergency physicians must maintain a high index of suspicion for infection, even in the face of nonspecific symptoms, and should tailor empiric antibiotic therapies to the expected pathogens in this population. Appropriate emergency department evaluation and treatment of infections in the elderly requires consideration of these unique aspects of the geriatric patient population. PMID- 18406978 TI - Infections related to pregnancy. AB - Infections in pregnancy and in the postpartum period represent an important knowledge base that emergency physicians should have. Being able to understand the risk that pregnancy-related infections have on maternal and fetal health in addition to the initiation of appropriate treatment for these infections is crucial. In the postpartum period, most infections (94%) are likely to manifest after hospital discharge; therefore, patients frequently present to the emergency department. PMID- 18406979 TI - HIV infection and complications in emergency medicine. AB - HIV infection is a common worldwide public health challenge affecting an estimated 40 million persons. In the United States, there have been over 900,000 cases of AIDS, and it is estimated that there are currently over 1 million HIV infected persons in the United States. Although the population of HIV seropositivity is concentrated in large urban settings, infections have been documented in all states. Because of the widespread prevalence and serious health consequences, it is imperative that the emergency physician be knowledgeable and skilled to diagnose and manage HIV-related emergency conditions. Knowledge of clinical presentations, differential diagnosis, early treatment strategies, and disposition options is crucial to the effective emergency department management of HIV infections and AIDS. PMID- 18406980 TI - Pneumonia in the emergency department. AB - Pneumonia remains one of the most common reasons for admission of emergency department (ED) patients to the hospital. Pneumonia also remains one of the most common causes of death in our patients. As with many emergent conditions, the ED management of these patients initiated by ED physicians contributes greatly to the survival and successful management of these patients. Specifically, the recognition of severe pneumonias, precise choice of diagnostic tests, and appropriate antibiotics can have an impact on the outcome. PMID- 18406981 TI - Urinary tract infections: diagnosis and management in the emergency department. AB - With the emergence of increasing resistance to common antibiotics used to treat urinary tract infections (UTIs), including ciprofloxacin and trimethoprim sulfamethoxazole (TMP-SMX), the choice of antibiotics for these infections has become more challenging. In this article, the authors review the evidence-based guidelines for the evaluation and treatment of cystitis and pyelonephritis in the emergency department. They review the pathophysiology and describe the initial diagnostic workup, spending some time discussing the urine dipstick. The authors discuss whether hospital antibiograms are useful in making the initial antibiotic choice. The treatment section reviews the current recommendations and also highlights the use of nitrofurantoin in the treatment of uncomplicated UTIs. The authors also discuss the appropriate use of ciprofloxacin and TMP-SMX in the treatment of UTIs. PMID- 18406982 TI - Community-associated methicillin-resistant Staphylococcus aureus. AB - Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has emerged over the last decade across the United States and the world, becoming a major pathogen in many types of community-acquired infections. Although most commonly associated with minor skin and soft tissue infections, such as furuncles, CA-MRSA also can cause necrotizing fasciitis, pyomyositis, osteoarticular infections, and community-acquired pneumonia. This article discusses the epidemiology, diagnosis, and management of these infections from the perspective of the emergency physician. PMID- 18406983 TI - Management of oral and genital herpes in the emergency department. AB - The epidemiology of oral and genital herpes has dramatically changed over the past decade. Herpes simplex virus-1, traditionally associated with oral herpes, is now implicated in an increasing percentage of genital herpes cases. The possibility of "autoinoculation" (or self-infection) of anatomic sites other than that of the primary infection has been recognized. New methods of suppression therapy are being examined. These changes have led to a revision in the recommendations by the Centers for Disease Control and Prevention (CDC). This review discusses herpes infections of the oral and genital mucosa and the suggested approach to the infected patient who presents in the emergency department. Specific attention is given to the CDC's 2006 guidelines for the treatment of sexually transmitted diseases. PMID- 18406984 TI - Foodborne illness. AB - While few patients with foodborne illness present with life-threatening symptoms, there are a number of foodborne infectious diseases and toxins that the emergency physician or other health care provider must consider in the evaluation of these patients. Given the frequency of international travel, as well as the risk associated with recurrent outbreaks of foodborne illness from commercial food sources, it is important to recognize various syndromes of foodborne illness, including those which may require specific evaluation and management strategies. This article reviews a number of the most common causes of foodborne illness, as well as several less common pathogens with the potential for causing significant morbidity and mortality if not promptly identified and treated. PMID- 18406985 TI - Travel-related infections. AB - Infections in travelers returning from international destinations are a common problem for emergency physicians. A careful travel history can help to distinguish the traveler's risk of having contracted an exotic infection, including malaria, dengue fever, and typhoid fever. The most common travel related infection is traveler's diarrhea. A discussion of typical and rare conditions is provided, grouped by the three most common chief complaints of fever, diarrhea, and rash. PMID- 18406986 TI - Infectious agents of bioterrorism: a review for emergency physicians. AB - The terrorist attacks on the United States in 2001 and the anthrax release soon after brought the issue of bioterrorism to the forefront in the medical community. Bioterrorism is the use of a biologic weapon to create terror and panic. Biologic weapons, or bioweapons, can be bacteria, fungi, viruses, or biologic toxins. Because the emergency department represents the front line of defense for the recognition of agents of bioterrorism, it is essential that emergency physicians have the ability to quickly diagnose victims of bioterrorism. This review examines the most deadly and virulent category A agents of bioterrorism, that is, anthrax, smallpox, plague, botulism, hemorrhagic fever viruses, and tularemia. The focus is on epidemiology, transmission, clinical manifestations, diagnosis, and treatment. PMID- 18406987 TI - Influenza and pneumococcal vaccinations in the emergency department. AB - Influenza and pneumococcal pneumonia remain among the most significant causes of morbidity and mortality of any of the infectious disease emergencies presenting to emergency departments (EDs). Because the ED has become a recommended location at which immunizations have been administered to prevent several infections, pneumococcal and influenza vaccinations can have an impact on the care of ED patients. ED personnel are uniquely positioned to vaccinate a substantial number of patients who would not otherwise be vaccinated, including many high-risk populations. In addition to decreasing vaccine-preventable mortality and morbidity from influenza and pneumococcal diseases, EDs that implement and monitor a systematic approach to these vaccinations can attenuate ED overcrowding and facilitate patient flow. ED vaccination strategies have been proved to be successful and reimbursable and are advocated by several major clinical practice advisory groups. PMID- 18406988 TI - Hyperbaric oxygen: applications in infectious disease. AB - This article reviews the applications of hyperbaric oxygen (HBO) as an adjunctive treatment of certain infectious processes. Infections for which HBO has been studied and is recommended by the Undersea and Hyperbaric Medicine Society include necrotizing fasciitis, gas gangrene, chronic refractory osteomyelitis (including malignant otitis externa), mucormycosis, intracranial abscesses, and diabetic foot ulcers that have concomitant infections. In all of these processes, HBO is used adjunctively along with antimicrobial agents and aggressive surgical debridement. This article describes the details of each infection and the research that supports the use of HBO. PMID- 18406990 TI - Thromboembolic disease and antithrombotic agents in the elderly. Preface. PMID- 18406991 TI - Heparins, low-molecular-weight heparins, and pentasaccharides: use in the older patient. AB - Age-related alterations in renal function, protein binding, and increased bleeding risk must be considered prior to administering anticoagulants to the increasing elderly population. Clinical use of unfractionated heparin, low molecular-weight heparin, and fondaparinux are reviewed with respect to their role in prevention and treatment of venous thromboembolism, as well as for treatment of acute coronary syndrome. In addition, heparin-induced thrombocytopenia, a potentially life-threatening adverse effect, is discussed with respect to both diagnosis and management. PMID- 18406992 TI - Warfarin pharmacology, clinical management, and evaluation of hemorrhagic risk for the elderly. AB - Warfarin, a vitamin K antagonist, is currently the most extensively used oral anticoagulant world-wide. It is prescribed for a variety of indications and has undergone extensive clinical study. Still, despite wide usage and considerable accumulated data from clinical trials demonstrating efficacy for a variety of thrombotic and thromboembolic conditions, warfarin is underutilized because its management is complex for both patients and physicians. However, despite these limitations, warfarin can be managed with relative safety, even in an elderly population. PMID- 18406993 TI - New anticoagulant agents: direct thrombin inhibitors. AB - Decades of research have been devoted to developing effective, safe, and convenient anticoagulant agents. In recent years, much emphasis has been placed on the development of direct thrombin inhibitors (DTIs) that offer benefits over agents like heparin and warfarin including the inhibition of both circulating and clot-bound thrombin; a more predictable anticoagulant response, because they do not bind to plasma proteins and are not neutralized by platelet factor 4; lack of required cofactors, such as antithrombin or heparin cofactor II; inhibiting thrombin-induced platelet aggregation; and absence of induction of immune mediated thrombocytopenia. Various injectable DTIs are currently available and used for many indications. In addition, research is now focusing on oral DTIs that seem promising and offer various advantages, such as oral administration, predictable pharmacokinetics and pharmacodynamics, a broad therapeutic window, no routine monitoring, no significant drug interactions, and fixed-dose administration. PMID- 18406994 TI - Antiplatelet agents and arterial thrombosis. AB - There is an increase in arterial thrombotic events in the elderly. Elderly patients are more likely to have associated diseases, such as diabetes, hypertension and hypercholesterolemia, and when age is confounded by these other predisposing factors, the risk of an arterial ischemic event increases disproportionately. Antithrombotic therapy for geriatric patients is underused, even when one adjusts for potential drug contraindications. This article focuses on the action of the currently available antiplatelet agents--aspirin, clopidogrel, and glycoprotein IIb/IIIa (GPIIb/IIIa) receptor antagonists, and assesses their effects in different disease states, with special attention to data that examine the geriatric population. PMID- 18406995 TI - Pathophysiology of venous thrombosis and the diagnosis of deep vein thrombosis pulmonary embolism in the elderly. AB - Deep vein thrombosis and pulmonary embolism are common medical problems in a variety of patient populations. It is essential that practitioners involved in the management of these patients understand not only the pathophysiology but also the diagnosis of venous thromboembolism. This foundation will provide the basis for the best care of this disease entity. PMID- 18406996 TI - Prevention of venous thromboembolism in the geriatric patient. AB - Elderly patients immobilized because of an acute medical illness or surgery have a very high risk of developing venous thromboembolism (VTE). Aggressive pharmacologic prophylaxis is necessary and should be initiated either at admission for a medical condition or shortly after surgery. Aggressive prophylaxis may result in fewer patients developing VTE in the hospital and ultimately lead to fewer patients requiring full-dose anticoagulation for VTE. Mechanical prophylaxis can be used as an adjunct to an anticoagulant-based regimen but should only be used as primary prophylaxis when there is a contraindication, such as active bleeding. It is recommended that the clinician carefully evaluate the elderly patient's creatinine clearance and weight before prescribing anticoagulants, particularly when using fixed dosing regimens. PMID- 18406997 TI - Anticoagulant treatment of deep vein thrombosis and pulmonary embolism. AB - Venous thrombosis is a common disease. As the mean age of the population increases, so does the incidence of venous thromboembolism. Anticoagulant therapy is equally effective in young and older patients, and can reduce substantially the associated morbidity and mortality. When considering long-term oral anticoagulant therapy in older patients, however, careful ongoing evaluation is imperative to ensure that the risk of bleeding does not outweigh the antithrombotic benefits. PMID- 18406998 TI - Antithrombotic and thrombolytic therapy for ischemic stroke. AB - Thrombolytic and antithrombotic agents form the cornerstone of stroke treatment and prevention. Recombinant tissue plasminogen activation (rt-PA) improves the outcome in patients treated within 3 hours of stroke onset. The risk-benefit ratio is narrow because of an increased risk for bleeding, but studies do not support a higher risk in the geriatric population. Emerging trials are directed at extending the therapeutic window and identifying agents that could provide better safety profiles. Large randomized trials have also highlighted the effectiveness and safety of early and continuous antiplatelet therapy in reducing atherothrombotic stroke recurrence. Aspirin has become the antiplatelet treatment standard against which several other antiplatelet agents have been shown to be more effective. The prevention of cardioembolic stroke is best accomplished with oral anticoagulation, barring any contraindications. PMID- 18406999 TI - Reducing the risk for stroke in patients who have atrial fibrillation. AB - Warfarin is highly effective at reducing the risk of stroke in AF. The benefit of oral anticoagulant therapy strongly outweighs the risk in most patients who have AF. More data are needed to define the overall risk-to-benefit ratio better for patients aged 80 years and older. Because a significant proportion of elderly individuals may not be optimal candidates for anticoagulant therapy, we must continue to evaluate alternative stroke prevention strategies while redoubling our efforts to understand the mechanisms underlying AF and thrombogenesis. PMID- 18407000 TI - Chronic antithrombotic therapy in post-myocardial infarction patients. AB - Because 1.1 million myocardial infarctions occur in the United States alone each year, and 450,000 of them are recurrent infarctions, which carry an inherently greater risk of death and disability than first events, the importance of secondary prevention strategies that can be implemented widely is unparalleled in health care. Antithrombotic therapies, both antiplatelet and anticoagulant, have become the mainstays of these strategies. This article covers the use of chronic antiplatelet and anticoagulation agents after myocardial infarction. It does not include the management of these patients in the acute phase. PMID- 18407001 TI - Antithrombotic therapy in peripheral arterial disease. AB - The management of elderly patients with peripheral arterial disease requires a multidisciplinary and individualized approach, especially for patients requiring intervention and for those on antithrombotic therapy. Communication between the patient's primary physician, consulting medical specialists, and vascular surgeon is essential because all may contribute synergistically to deliver optimal care to the patient. This article reviews the pathophysiology of peripheral arterial disease and data regarding the use of antiplatelet and anticoagulant agents. PMID- 18407002 TI - Perioperative management of oral anticoagulation. AB - Based on an individual assessment of risk factors for arterial or venous thrombosis and the risk of postoperative bleeding, this article outlines the preoperative and postoperative approach to anticoagulant management. Preceding this is a brief description of the therapies most commonly used in the perioperative period. The prevention of arterial thromboembolism is considered separately from the prevention of venous thrombosis. Perioperative management of anticoagulation can cause anxiety for patients, surgeons, anesthetists, and those who manage long-term anticoagulant therapy. Good communication between all of these parties is essential to ensure that an optimal management strategy is identified and executed. PMID- 18407003 TI - Web-based smoking-cessation programs: results of a randomized trial. AB - BACKGROUND: Initial trials of web-based smoking-cessation programs have generally been promising. The active components of these programs, however, are not well understood. This study aimed to (1) identify active psychosocial and communication components of a web-based smoking-cessation intervention and (2) examine the impact of increasing the tailoring depth on smoking cessation. DESIGN: Randomized fractional factorial design. SETTING: Two HMOs: Group Health in Washington State and Henry Ford Health System in Michigan. PARTICIPANTS: 1866 smokers. INTERVENTION: A web-based smoking-cessation program plus nicotine patch. Five components of the intervention were randomized using a fractional factorial design: high- versus low-depth tailored success story, outcome expectation, and efficacy expectation messages; high- versus low-personalized source; and multiple versus single exposure to the intervention components. MEASUREMENTS: Primary outcome was 7 day point-prevalence abstinence at the 6-month follow-up. FINDINGS: Abstinence was most influenced by high-depth tailored success stories and a high personalized message source. The cumulative assignment of the three tailoring depth factors also resulted in increasing the rates of 6-month cessation, demonstrating an effect of tailoring depth. CONCLUSIONS: The study identified relevant components of smoking-cessation interventions that should be generalizable to other cessation interventions. The study also demonstrated the importance of higher-depth tailoring in smoking-cessation programs. Finally, the use of a novel fractional factorial design allowed efficient examination of the study aims. The rapidly changing interfaces, software, and capabilities of eHealth are likely to require such dynamic experimental approaches to intervention discovery. PMID- 18407005 TI - Respiratory disease in Army recruits: surveillance program overview, 1995-2006. AB - BACKGROUND: Since 1966, the U.S. Army has had in place a comprehensive surveillance system for acute respiratory disease (ARD) because of its continued high incidence in the recruit population. A retrospective review of the program from 1995 to 2006, a continuation of an earlier analysis from 1985-1994, was performed to describe the program and to summarize group A streptococcal (GAS) outbreaks identified through this program. METHODS: Weekly incidence data and daily sentinel case data collected as part of the ARD surveillance program were analyzed in 2007 to report rates of ARD and GAS outbreaks over this 12-year period (1995-2006) for five participating Army sites. RESULTS: Average ARD rates were consistently higher from 2000 to 2006 than from 1995 to 1999. Rate ratios of winter rates to summer rates were greater than 1.0 prior to 2000, but from 2000 to 2006, ratios were consistently near 1.0. Rheumatogenic M types were available for three of five outbreaks. CONCLUSIONS: Continuing high incidence of ARD in the military, re-emerging rheumatogenic M types, and a highly mobile military underscore the need for continued surveillance efforts. The continued employment of interventions with demonstrated effectiveness, such as chemoprophylaxis and basic preventive measures, is essential to reducing the burden of ARD. PMID- 18407004 TI - Effect of incentives and mailing features on online health program enrollment. AB - BACKGROUND: With the growing use of Internet-based interventions, strategies are needed to encourage broader participation. This study examined the effects of combinations of monetary incentives and mailing characteristics on enrollment, retention, and cost effectiveness for an online health program. METHODS: In 2004, a recruitment letter was mailed to randomly selected Midwestern integrated health system members aged 21-65 and stratified by gender and race/ethnicity; recipients were randomly pre-assigned to one of 24 combinations of incentives and various mailing characteristics. Enrollment and 3-month retention rates were measured by completion of online surveys. Analysis, completed in 2005, compared enrollment and retention factors using t tests and chi-square tests. Multivariate logistic regression modeling assessed the probability of enrollment and retention. RESULTS: Of 12,289 subjects, 531 (4.3%) enrolled online, ranging from 1% to 11% by incentive combination. Highest enrollment occurred with unconditional incentives, and responses varied by gender. Retention rates ranged from 0% to 100%, with highest retention linked to higher-value incentives. The combination of a $2 bill prepaid incentive and the promise of $20 for retention (10% enrollment and 71% retention) was optimal, considering per-subject recruitment costs ($32 enrollment, $70 retention) and equivalent enrollment by gender and race/ethnicity. CONCLUSIONS: Cash incentives improved enrollment in an online health program. Men and women responded differently to mailing characteristics and incentives. Including a small prepaid monetary incentive ($2 or $5) and revealing the higher promised-retention incentive was cost effective and boosted enrollment. PMID- 18407006 TI - Adverse childhood experiences and chronic obstructive pulmonary disease in adults. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is an important cause of morbidity and mortality in the U.S. However, little is known about the influence of childhood stressors on its occurrence. METHODS: Data were from 15,472 adult HMO members enrolled in the Adverse Childhood Experiences (ACE) Study from 1995 to 1997 and eligible for the prospective phase. Eight ACEs were assessed: abuse (emotional, physical, sexual); witnessing domestic violence; growing up with substance-abusing, mentally ill, or criminal household members; and parental separation or divorce. The number of ACEs (ACE Score) was used to examine the relationship of childhood stressors to the risk of COPD. Three methods of case ascertainment were used to define COPD: baseline reports of prevalent COPD, incident hospitalizations with COPD as a discharge diagnosis, and rates of prescription medications to treat COPD during follow-up. Follow-up data were available through 2004. RESULTS: The ACE Score had a graded relationship to each of three measures of the occurrence of COPD. Compared to people with an ACE Score of 0, those with an ACE Score of > or =5 had 2.6 times the risk of prevalent COPD, 2.0 times the risk of incident hospitalizations, and 1.6 times the rates of prescriptions (p<0.01 for all comparisons). These associations were only modestly reduced by adjustment for smoking. The mean age at hospitalization decreased as the ACE Score increased (p<0.01). CONCLUSIONS: Decades after they occur, adverse childhood experiences increase the risk of COPD. Because this increased risk is only partially mediated by cigarette smoking, other mechanisms by which ACEs may contribute to the occurrence of COPD merit consideration. PMID- 18407007 TI - Racial and ethnic disparities in smoking-cessation interventions: analysis of the 2005 National Health Interview Survey. AB - BACKGROUND: Recent national surveys document racial and ethnic disparities in receipt of smoking-cessation advice. This study updates and expands prior analyses using survey data for 2005, and evaluates the association between smokers' race and ethnicity and three separate measures of healthcare-encounter based tobacco interventions: screening, smoking-cessation advice, and use of smoking-cessation aids. METHODS: Analyses are based on 4756 smokers (aged 18 and older) reporting a healthcare encounter within the past year who participated in the 2005 National Health Interview Survey (NHIS). Multivariate-adjusted OR and 95% CI for receipt of tobacco interventions in non-Hispanic black and Hispanic smokers were compared to those of non-Hispanic white smokers, adjusted for smokers' characteristics (sociodemographics, health status, and healthcare utilization factors, and smoking-related characteristics). Analyses were done in 2006. RESULTS: Results show that compared to white smokers, black and Hispanic smokers had significantly lower odds of (1) being asked about tobacco use (AOR=0.70 and AOR=0.69, respectively); (2) being advised to quit (AOR=0.72 and AOR=0.64, respectively); or (3) having used tobacco-cessation aids during the past year in a quit attempt (AOR=0.60 and AOR=0.59, respectively). Compared to 2000 NHIS published data, the prevalence of receipt of advice to quit from a healthcare provider increased from 52.9% in 2000 to 61.2% in 2005, with increases across racial and ethnic groups. CONCLUSIONS: Despite progress in smokers' being advised to quit during healthcare encounters in the past 5 years, black and Hispanic smokers continue to be less likely than whites to receive and use tobacco-cessation interventions, even after control for socioeconomic and healthcare factors. Further actions are needed to understand and eliminate this disparity. PMID- 18407008 TI - Physical activity and neighborhood resources in high school girls. AB - BACKGROUND: Physical activity behavior is influenced by a person's physical environment, but few studies have used objective measures to study the influences of the physical environment on physical activity behavior in youth. The purpose of this study was to examine the relationship between selected neighborhood physical activity resources and physical activity levels in high school girls. METHODS: Participants were students in schools that had participated in a large physical activity intervention trial. The 3-Day Physical Activity Recall was completed by 1506 12th-grade girls. Data on physical activity facilities and resources in the participating communities were collected using a variety of methods. Physical activity resources within a 0.75-mile street-network buffer around each girl's home were counted using ArcGIS, version 9.1. Mixed-model regression models were used to determine if there was a relationship between three physical activity variables and the number of physical activity resources within the 0.75-mile buffer. Data were collected in 2002-2003 and analyzed in 2006-2007. RESULTS: On average, 3.5 physical activity resources (e.g., schools, parks, commercial facilities) were located within the 0.75-mile street-network buffer. Thirty-six percent of the girls had no physical activity resource within the buffer. When multiple physical activity resources were considered, the number of commercial physical activity facilities was significantly associated with reported vigorous physical activity, and the number of parks was associated with total METs in white girls. CONCLUSIONS: Multiple physical activity resources within a 0.75-mile street-network buffer around adolescent girls' homes are associated physical activity in those girls. Several types of resources are associated with vigorous physical activity and total activity in adolescent girls. Future studies should examine the temporal and causal relationships between the physical environment, physical activity, and health outcomes related to physical activity. PMID- 18407010 TI - Cigar/cigarillo surveillance: limitations of the U.S. Department of Agriculture system. AB - BACKGROUND: The U.S. Department of Agriculture (USDA) reports data on cigar sales in two categories: little cigars (weighing less than 3 lbs. per thousand) and large cigars and cigarillos (weighing more than 10 lbs. per thousand). A rise in the sales of little cigars in recent years is a cause for concern. The capacious second category could be obscuring the growth of sales in cigarillos. METHODS: Trends in cigar use were analyzed in May 2007 using (1) the standard USDA two level system and (2) data from the Maxwell Report that provides information on cigarillos as a separate category. The intercorrelations among cigar use trends in the three areas were also explored. RESULTS: From 1993 to 2006, unit sales of little cigars increased from 37% to 47% of the cigar market, cigarillos increased from 25% to 32%, and large cigars dropped from 37% to 22%. From 1976 to 2006, cigarillo sales were strongly related to sales of little cigars (r=0.93; 95% CI=0.86-0.97), while sales of large cigars and cigarillos were modestly related (r=0.42; 95% CI=0.08-0.57). CONCLUSIONS: Analyses show strong correlations between cigarillo and little cigar sales and argue for more detailed reporting of cigar sales as a function of cigar size. Tobacco surveillance should at minimum be watching the same trends as the tobacco industry. The sales of little cigars, cigarillos, and large cigars should be monitored, and the measuring of cigarillo sales in 3-5 subcategories according to size is encouraged. PMID- 18407009 TI - Injury reporting on local TV news: a prime-time opportunity for prevention. AB - BACKGROUND: Local television news is America's primary source of information and may be an opportunity to shape public opinion surrounding issues such as injury prevention. OBJECTIVE: This study sought to systematically evaluate unintentional injury coverage on local television news and to identify frequently interviewed public-service professionals and factors associated with discussion of risk factors and prevention. METHODS: Late news broadcasts from 122 local television stations within the U.S. during October 2002 were analyzed. The main outcomes variables were counts of case-injury stories: motor-vehicle crashes, fires, falls, drowning, poisonings, and sports-recreational injuries; identification of interviewed public service professionals; and discussion of risk factors and prevention. Bivariate and mulitvariate analysis was performed to identify predictors of discussion of prevention measures, risk factors, or both. Data were analyzed in Fall 2006. RESULTS: From 2795 broadcasts, 1748 case-injury stories were identified. Fires and motor-vehicle crashes constituted 84% of the case injury stories. There were 245 case-injury stories containing an interview with a public service professional. Police officers and firefighters accounted for 82% of these interviews. Interviews with police officers and firefighters were independently associated with discussion of risk factors and prevention measures for motor-vehicle crashes (OR=2.49, CI=1.7-3.6) and fires (OR=2.77, CI=1.2-5.9), respectively. CONCLUSIONS: Motor-vehicle crashes and fires were the most commonly reported injury topics. Police officers and firefighters were most commonly interviewed and, if interviewed, increased the likelihood that risk factors, prevention measures, or both were discussed. Optimizing the messages delivered by public service professionals through public service professional-level and media level interventions may be an opportunity for disseminating injury-prevention information to the public and to policymakers, and methods to increase the likelihood of media interviews with public service professionals should be explored. PMID- 18407011 TI - Shifting schedules: the health effects of reorganizing shift work. AB - BACKGROUND: Approximately one fifth of workers are engaged in some kind of shift work. The harmful effects of shift work on the health and work-life balance of employees are well known. A range of organizational interventions has been suggested to address these negative effects. METHODS: This study undertook the systematic review (following Quality Of Reporting Of Meta [QUORUM] analyses guidelines) of experimental and quasi-experimental studies, from any country (in any language) that evaluated the effects on health and work-life balance of organizational-level interventions that redesign shift work schedules. Twenty seven electronic databases (medical, social science, economic) were searched. Data extraction and quality appraisal were carried out by two independent reviewers. Narrative synthesis was performed. The review was conducted between October 2005 and November 2006. RESULTS: Twenty-six studies were found relating to a variety of organizational interventions. No one type of intervention was found to be consistently harmful to workers. However, three types were found to have beneficial effects on health and work-life balance: (1) switching from slow to fast rotation, (2) changing from backward to forward rotation, and (3) self scheduling of shifts. Improvements were usually at little or no direct organizational cost. However, there were concerns about the generalizability of the evidence, and no studies reported on impacts on health inequalities. CONCLUSIONS: This review reinforces the findings of epidemiologic and laboratory based research by suggesting that certain organizational-level interventions can improve the health of shift workers, their work-life balance, or both. This evidence could be useful when designing interventions to improve the experience of shift work. PMID- 18407012 TI - Preschool children and physical activity: a review of correlates. AB - BACKGROUND: Several reviews have summarized the research on correlates of older children's and adolescents' physical activity behaviors, but none have been published on preschool children. Over the past 27 years, a number of studies have investigated the correlates of preschool children's physical activity behaviors. It is timely and necessary to review the extant literature in this area. This paper reviews articles investigating correlates of preschool children's physical activity behaviors published in peer-reviewed journals between 1980 and March 2007. METHODS: A literature search was conducted to identify studies that investigated correlates of preschool children's physical activity. Data were collected and analyzed in 2007. RESULTS: Twenty-four articles were identified that met the inclusion criteria. From those articles, 39 variables were identified across five domains. Results showed that boys were more active than girls, that children with active parents tended to be more active, and that children who spent more time outdoors were more active than children who spent less time outdoors. Age and BMI were consistently shown to have no association with preschool children's physical activity. Other variables produced largely inconclusive results. CONCLUSIONS: The influences on the physical activity behaviors of preschool children are multidimensional. Further research is required to enhance an understanding of these influences. PMID- 18407013 TI - Clinical skills and self-efficacy after a curriculum on care for the underserved. AB - BACKGROUND: Despite calls for medical school curricula that address care for the underserved, published evaluations of such curricula are few and often do not assess clinical skills. This study assesses the changes in self-efficacy and clinical skills resulting from faculty-led or web-based curricula on care for the underserved. METHODS: Third-year students on 6-week 2003-2004 pediatric clerkships were block-randomized to a curriculum on caring for the underserved in one of three formats: established (readings only), faculty-led, or web-based. Primary outcomes were pre- and post-curriculum changes in self-efficacy and clinical skills. ANCOVA was used to test differences among curriculum groups, adjusting for prior experiences with the underserved. Analyses were performed in 2007. RESULTS: A total of 138 students participated, with 121 (88%) completing both pre- and post-tests. Compared to the established-curriculum students, both faculty-led and web-based students improved their self-efficacy in establishing achievable goals with underserved families, while web-based students improved their self-efficacy around knowledge of community resources. Significantly more new skills were performed by both faculty-led (mean [SD]=3.3[1.8]) and web-based curriculum students (2.9[1.5]), compared to established-curriculum students (1.5[1.4]). Compared to established-curriculum students, more faculty-led and web based students also reported new skills in locating resources to meet the needs of underserved patients and in following up on referrals to ensure that families accessed needed care. CONCLUSIONS: Web-based and faculty-led curricula improve medical student self-efficacy and clinical skills. Results from specific self efficacy and skill items facilitate targeted curricular improvement. PMID- 18407014 TI - Answering the "what works?". Question in health behavior change. PMID- 18407015 TI - Translation of the 2002 hormone study findings. PMID- 18407016 TI - In-vivo evaluation of the contamination of Super Slick elastomeric rings by Streptococcus mutans in orthodontic patients. AB - INTRODUCTION: We investigated in vivo the contamination by Streptococcus mutans of Super Slick elastomeric rings (TP Orthodontics, LaPorte, Ind), manufactured with Metafasix technology (TP Orthodontics), using microbial culture and scanning electron microscopy (SEM). METHODS: Twenty patients undergoing fixed orthodontic appliance therapy were selected. Super Slick elastomeric rings (n = 160) were tied to brackets on the right maxillary premolars or molars and left mandibular premolars or molars. Conventional elastomeric rings (n = 160) were tied to brackets on the contralateral premolars or molars with the same split-mouth design. After a 15-day intraoral period, 75 elastomeric rings of each type were retrieved, submitted to microbiologic processing, and cultured in bacitracin sucrose broth-selective enrichment broth culture media. The number of S mutans colonies or biofilms on the surface of the electrometric rings was counted by using a stereomicroscope. Data were analyzed statistically with the Wilcoxon nonparametric test at the 5% significance level. Four representative rings of each type were chosen for SEM analysis. RESULTS: Statistical analysis by the Wilcoxon nonparametric test showed that the Super Slick elastomeric rings had statistically significant greater S mutans contamination than the conventional elastomeric rings (P <.0001). No formation of S mutans colonies or biofilms was observed in the elastomeric rings removed directly from their original packages. SEM micrographs showed fissures on the surface of Super Slick elastomeric rings. No fissures were found on conventional elastomeric rings. When the microbiologic culture was positive, S mutans bacterial biofilm was observed on both types of ligatures. CONCLUSIONS: There was no clinical evidence that Super Slick elastomeric rings are effective in reducing bacterial biofilm formation on their surfaces, and a recommendation for their use in orthodontic therapy for that purpose is not justifiable. PMID- 18407017 TI - Soft-tissue lasers in orthodontics: an overview. AB - Soft-tissue lasers have numerous applications in orthodontics, including gingivectomy, frenectomy, operculectomy, papilla flattening, uncovering temporary anchorage devices, ablation of aphthous ulcerations, exposure of impacted teeth, and even tooth whitening. As an adjunctive procedure, laser surgery has helped many orthodontists to enhance the design of a patient's smile and improve treatment efficacy. Before incorporating soft-tissue lasers into clinical practice, the clinician must fully understand the basic science, safety protocol, and risks associated with them. The purpose of this article is to provide an overview regarding safe and proper use of soft-tissue lasers in orthodontics. PMID- 18407018 TI - Demographic characteristics of orthodontic practice sites in the United States. AB - Few studies have addressed the demographic characteristics of dental-practice sites, including orthodontic practice sites. In this study, we investigated the demographic characteristics of orthodontic practice sites in the United States. Demographic data and orthodontic practices were organized by zip codes and analyzed with discriminant analysis. The demographic characteristics associated with zip codes that contained an orthodontic practice were determined vs those without an orthodontic practice. The resulting model correctly classified 90.3% of the 30,134 zip code areas, based on the presence or absence of an orthodontic practice. The variables most closely associated with a zip code containing an orthodontic practice included number of dental practices (general practice) and population size. Several variables related to the socioeconomic status of the area were also positively related to zip codes with an orthodontic practice. Further analysis indicated a positive relationship between the model's output and its ability to predict single vs several orthodontic practices in a zip code area. A practical method for assessing the relative merits of alternate site locations is also provided. PMID- 18407019 TI - Treatment of severe Class II Division 1 deep overbite malocclusion without extractions in an adult. AB - This case report describes the orthodontic treatment and retention of an adult with severe Class II Division 1 deep overbite malocclusion; treatment was completed without premolar extractions. We used improved superelastic nickel titanium alloy wire (L&H Titan, Tomy, Tokyo, Japan) with J-hook headgear for the combined intrusion of the anterior segment and the distalization of the maxillary posterior segments. The overbite was corrected from 6.0 to 2.0 mm by intrusion of the maxillary and mandibular incisors; the overjet was corrected from 13.0 to 2.0 mm. Although no premolars were extracted, good occlusion and facial esthetics were achieved. Follow-up records 2 years after active treatment show that the results were maintained. PMID- 18407020 TI - A new computer-assisted method for design and fabrication of occlusal splints. AB - In this report, we describe computer-based design and production of occlusal splints. A research effort was undertaken to develop a process to eliminate the inherent variabilities associated with current splint-fabrication methods. The digital process provides quantitative control over articulation and splint design, and produces splints with continuously smooth occlusal surfaces. Stone casts are laser scanned, and custom software is used to articulate and design flat-plane and full-coverage splints with guidance ramps. Splints are produced by milling excess acrylic placed over stone casts. Clinically, digital splints reduce the average time needed for placement because intraoral equilibration is minimized. PMID- 18407022 TI - Factors affecting information retention in orthodontic patients. AB - INTRODUCTION: Our purpose was to determine the factors that affect patients' retention of information provided in different formats. METHODS: Eighty new patients attending clinics were randomly allocated into 1 of 2 groups. The first group was given a commercial information leaflet about orthodontic treatment, and the second group saw a visual computer program with the same information. Both groups were given additional verbal information. Short- and longer-term retention of information were assessed using a questionnaire. Age, sex, ethnicity, index of relative deprivation of area of residence, and the time taken to view or read the information were recorded. Linear regression analysis was undertaken to assess the relationship between retention of information and these variables. RESULTS: The response rates for the first and second questionnaires were 100% and 67.5%, respectively. The method of information provision was found to be the only statistically significant factor affecting information retention, with the visual computer program group achieving higher scores than the written information leaflet group. CONCLUSIONS: Computer-based visual information was shown to be a better method for information retention. Consequently, it is worth considering providing information to orthodontic patients in a more visual format if it is to be retained most effectively. PMID- 18407024 TI - A new method of assessing aerosolized bacteria generated during orthodontic debonding procedures. AB - INTRODUCTION: The main objective of this study was to assess the efficacy of a new and innovative method of harvesting bacteria that are aerosolized during orthodontic debonding. Additionally, the protection efficacy of several commercially available masks from such aerosols was assessed in a pilot study. METHODS: Twenty-six subjects were debonded during aerosol sampling, by using an innovative collection system to harvest bonding dust liberated during debonding. Dark-field microscopy, gram-stain microscopy, and chemical identification were used to determine speciation of the collected aerosol from 23 subjects. Three additional subjects were used to test 3 commercial dental or protective masks to determine whether they provide effective protection from the aerosol. RESULTS: Twenty-one species of oral bacteria were identified by the new sampling technique. Two of the 3 masks that were tested offered no protection against the aerosolized bacteria. CONCLUSIONS: A new and effective method for collecting airborne bacteria is presented. Some conventional dental masks offer no protection from aerosolized organisms liberated during debonding procedures. Further assessment of mask efficacy is ongoing. PMID- 18407023 TI - Assessment of perceived orthodontic appliance attractiveness. AB - INTRODUCTION: To date, no studies have been published in the literature evaluating the esthetics of orthodontic appliances. The purpose of this study was to assess appliance variables, including appliance type, brand, wire, and ligature tie, for their relative attractiveness. Demographic factors were also evaluated. METHODS: Appliances were placed in an adult, and digital images were captured, standardized, and incorporated into a computer-based survey. Subjects (n = 200) rated each image for attractiveness on a visual analog scale. The attractiveness ratings were analyzed by using factorial analysis of variance with repeated measures. Post-hoc testing was done with the Tukey-Kramer test (overall a <.05). RESULTS: For all demographic factors, attractiveness ratings were significantly different between alternative (clear trays and simulated lingual appliances), ceramic, and all self-ligating/stainless steel appliances. No differences between brands in each appliance type were found. Ratings of some ceramic appliances were significantly affected by archwire and ligature tie, depending on several demographic interactions. CONCLUSIONS: Orthodontic appliance attractiveness varies by the following hierarchy of appliance types: alternative > ceramic > all stainless steel/self-ligating. Wire and tie selection can affect the appearance of ceramic appliances but have no impact on stainless steel or self-ligating appliances. Certain demographic factors have variable effects on attractiveness ratings. PMID- 18407025 TI - Assessment of perceived orthodontic appliance attractiveness. PMID- 18407026 TI - Inhibition of enamel demineralization by an enamel sealant, Pro Seal: an in-vitro study. AB - INTRODUCTION: Enamel demineralization adjacent to fixed orthodontic appliances compromises both esthetics and oral health. The purpose of this in-vitro study was to evaluate the effectiveness of a new enamel sealant, Pro Seal (Reliance Orthodontic Products, Itasca, Ill), on inhibiting enamel demineralization. Two materials that have demonstrated success in white spot prevention and do not require patient compliance were used for comparison. METHODS: Thirty-two noncarious extracted molars were divided into 4 groups and received 1 of the following treatments: no treatment (control), fluoride varnish (Fluor Protector; Ivoclar Vivadent, Amherst, Mass), unfilled resin sealant (Delton; Dentsply Professional, York, Pa), and filled resin sealant (Pro Seal). The teeth were subjected to 15,000 simulated brush strokes followed by acidic challenge for 96 hours. They were examined macroscopically and sectioned for quantitative examination with polarized light microscopy. RESULTS: All surface treatments provided statistically significant (P <.05) reductions in lesion depth compared with controls. Pro Seal performed significantly better (P <.05) than the other products, decreasing lesion depth by 97% compared with the controls and completely inhibiting lesion formation in 3 specimens. CONCLUSIONS: Pro Seal shows promise as an effective method of preventing enamel demineralization without patient compliance. PMID- 18407027 TI - Effect of fluoride varnish on enamel demineralization around brackets: an in-vivo study. AB - INTRODUCTION: Even with advances in materials and techniques, demineralization around brackets during orthodontic treatment continues to be a problem. The purpose of this in-vivo study was to evaluate the effect of a fluoride varnish on enamel demineralization adjacent to bonded brackets. METHODS: Fifteen patients who needed at least 2 premolars extracted for orthodontic reasons were selected. In each patient, 1 premolar was considered the test tooth, and the other was the control. Brackets were bonded, and T-loops were engaged on all premolars, but only the test teeth received fluoride varnish. The premolars were extracted after 85 to 95 days, and buccolingual sections 50 to 70 microm in thickness were evaluated with polarized light microscopy. The mean depth of demineralization in each lesion was measured 3 times on photographs by an operator blinded to the groups (intraclass correlation of the 3 measurements was 0.988). RESULTS: The mean lesion depths were 57.0 +/- 5.5 microm in the test group and 94.3 +/- 6.7 microm in the control group. There was significant reduction (approximately 40%) in depth of demineralization in the test group (P <.001). CONCLUSIONS: Fluoride varnish can be beneficial as a preventive adjunct in reducing demineralization adjacent to brackets. PMID- 18407028 TI - Bacterial adhesion of Streptococcus mutans to esthetic bracket materials. AB - INTRODUCTION: The formation of white spot lesions and enamel demineralization might be intensified by enhanced bacterial adhesion and formation of dental plaque on brackets. Our aim in this in-vitro study was to investigate the susceptibility of various plastic bracket materials to the adherence of Streptococcus mutans. METHODS: The esthetic bracket raw materials included in this study were polyoxymethylene, polycarbonate, high-density polyethylene, and an experimental polymer (90% polyethylene). Surface roughness was assessed by perthometer. S mutans suspension was incubated with test specimens and examined with fluorescence dye and an automated multi-detection reader. The oxidation reduction fluorescence dye Alamar Blue/resazurin (0.007536 g per 10 mL distilled water) was used to determine the quantity of bacterial adhesion. RESULTS: The median values of fluorescence intensity varied between approximately 600 and 1600. The lowest fluorescence with median values below 700, indicating low bacterial adhesion, was found for polyoxymethylene (median of relative fluorescence intensity, 635). The highest relative fluorescence intensity, with median values of more than 1500, was for high-density polyethylene (1565). No statistical alteration of fluorescence intensity was measured between the polymeric bracket materials. CONCLUSIONS: The investigated polymeric bracket materials had no significant differences in the quantities of S mutans adhering to them. Regardless of the polymer, plastic bracket materials have similar bacterial colonization and plaque accumulation properties. PMID- 18407029 TI - Insulin therapy: evolution and practice. PMID- 18407030 TI - Our evolving understanding of getting to goal using insulin in type 2 diabetes. PMID- 18407031 TI - The effects of insulin on the endothelium. AB - Recent data suggest that diabetes in general, and particularly diabetes in association with insulin resistance, obesity, and hyperglycemia, results in inflammatory changes including the production of cytokines, adhesion molecules, and reactive oxygen species that are toxic to the endothelium and could lead to vascular damage. Insulin suppresses these effects, either indirectly by decreasing glucose levels or directly by stimulating nitric oxide production and inhibiting important pathways in the inflammatory cascade. In addition, insulin directly reduces plasma concentrations of adhesion molecule production in endothelial cells and thus may decrease vascular inflammation. Some of these effects of insulin may be attenuated in patients with insulin resistance, and this finding may provide a mechanistic link for the increase in vascular disease seen in patients with insulin resistance. Doses of insulin adequate to restore normoglycemia may help overcome these abnormalities, although some patients may require insulin sensitization with lifestyle changes or pharmacologic therapy. PMID- 18407032 TI - Fine-tuning glycemic control using computerized downloading software: a case based approach. AB - Diabetes self-management is not always the simplest of tasks. Patients with diabetes do not have a functioning pancreas. They rely on their knowledge to guide them through the complex decisions that must be made continually. Glycemic variability is not always a result of patient noncompliance. In fact, most patients are attempting to maintain normal glycemia. Imagine the frustrations that our patients feel when they do not understand why their blood glucose levels vary so greatly, even if they eat the same foods and participate in the same type of physical activities each day. As physicians, we should provide our patients with the best opportunities, tools, and technologies available to minimize their exposure to glycemic variability, oxidative stress, and long-term diabetes related complications. SBGM is an essential component in diabetes care. Incorporating computerized software into our practices so that we are able to fine-tune our prescribed treatments is an inexpensive and efficient way to improve the quality of our patients' lives. PMID- 18407033 TI - Use of continuous glucose monitoring to improve diabetes mellitus management. PMID- 18407034 TI - Future directions for insulin therapy and diabetes treatment. PMID- 18407035 TI - Irregular menses: a warning sign of vomiting for weight control. PMID- 18407036 TI - Health information on the internet--a double-edged sword? PMID- 18407037 TI - Hookahs and waterpipes: cultural tradition or addictive trap? PMID- 18407038 TI - Do young people's illness beliefs affect healthcare? A systematic review. AB - A successful patient-centered approach in clinical practice implies an understanding of patients' experiences of illness, and therefore of their illness beliefs. This paper presents a systematic review of the literature on young people's common illness beliefs in contemporary Western societies and the extent to which these beliefs have been shown to affect healthcare. PMID- 18407039 TI - Irregular menses linked to vomiting in a nonclinical sample: findings from the National Eating Disorders Screening Program in high schools. AB - PURPOSE: Using data from an eating disorders screening initiative conducted in high schools across the United States, we examined the relationship between vomiting frequency and irregular menses in a nonclinical sample of adolescent females. METHODS: A self-report questionnaire was administered to students from U.S. high schools participating in the National Eating Disorders Screening Program in 2000. The questionnaire included items on frequency of vomiting for weight control in the past 3 months, other eating disorder symptoms, frequency of menses, height, and weight. Multivariable regression analyses were conducted using data from 2791 girls to estimate the risk of irregular menses (defined as menses less often than monthly) associated with vomiting frequency, adjusting for other eating disorder symptoms, weight status, age, race/ethnicity, and school clusters. RESULTS: Girls who vomited to control their weight one to three times per month were one and a half times more likely (risk ratio [RR] = 1.6; 95% confidence interval [CI] = 1.2-2.2), and girls who vomited once per week or more often were more than three times more likely (RR = 3.2; 95% CI = 2.3-4.4), to experience irregular menses than were girls who did not report vomiting for weight control. Vomiting for weight control remained a strong predictor of irregular menses even when overweight and underweight participants were excluded. CONCLUSIONS: Our study adds to the evidence that vomiting may have a direct effect on hormonal function in adolescent girls, and that vomiting for weight control may be a particularly deleterious component of eating disorders. PMID- 18407040 TI - The availability and portrayal of stimulants over the Internet. AB - PURPOSE: To quantify the online availability and portrayal of amphetamine-class prescription stimulants with a focus on those medications commonly prescribed to and abused by adolescents. METHOD: The Google search engine was used in searches to assess the frequency of web sites offering to sell controlled stimulants (retail sites) or web sites that directly linked to retail sites (portal sites). In addition separate searches were used to evaluate the portrayal of controlled prescription stimulants by the initial 20 web sites returned by Google. Retail and portal web site frequency was collected for each search. For searches measuring the portrayal of stimulants, web pages were categorized as pro-use, anti-misuse, neutral or other, based on set criteria. RESULTS: Sites offering to sell stimulants without a prescription were found for nearly all search terms. Across all searches, the Schedule III stimulants indicated for the treatment of obesity returned more sites offering to sell stimulants without a prescription than Schedule II stimulants indicated for the treatment of attention-deficit hyperactivity disorder (ADHD). Internet site portrayal of each stimulant varied; however sites that contained "methamphetamine" often included anti-misuse information. CONCLUSIONS: The apparent availability of stimulants over the Internet without a prescription indicates the potential for a significant public health problem. The extent to which teens are obtaining these drugs via the Internet remains unclear, but clinicians must be aware of the potential for abuse, concomitant prescription use issues, illicit sources, and diversion of these medications, which can be highly addictive. Education of consumers and physicians as well as further governmental interventions are needed to limit the potential scope of this problem. PMID- 18407041 TI - Outcome of cognitive behavioral therapy in adolescents after natural disaster. AB - PURPOSE: The authors evaluated the effectiveness of cognitive behavioral therapy (CBT) among adolescents exposed to the 2004 earthquake in Bam, Iran. METHODS: Four months after the earthquake, 135 adolescents as a case group and 33 adolescents as a comparison group were evaluated with the Impact of Event Scale Revised (IES-R). Two therapists were trained in CBT in 3-day classes according to a manual provided by mental health services. After conducting CBT in the case group, both groups were evaluated again with IES-R. RESULTS: The severity of posttraumatic stress symptoms significantly decreased among the subjects given CBT in the case group. The improvement in posttraumatic stress symptoms was attributable to improvement in each of three-symptom categories (intrusion, avoidance, and arousal) and in the total score of posttraumatic stress disorder (p < .05). CONCLUSIONS: The findings demonstrate the efficacy of CBT in alleviating posttraumatic stress symptoms among adolescents after a catastrophic disaster. PMID- 18407042 TI - Gender differences in dating aggression among multiethnic high school students. AB - PURPOSE: (1) To assess prevalence of physical dating aggression and victimization among high school students; (2) to assess prevalence of mutual and exclusive aggression; (3) to determine whether aggression differs across ethnic groups and relationship type; and (4) to ascertain the likelihood of injury and breakup in individuals who reported that they were the recipients of physical aggression. METHODS: Students (N = 2363) from seven multiethnic high schools participated. Because males in high school date females younger than they and the reverse for females, and because males and females may underreport aggression, only within gender comparisons were conducted. RESULTS: More females reported engaging in physical aggression (40%) than reported being victims of aggression (30%). Fewer males reported engaging in physical aggression (24%) than reported being victims of physical aggression (31%). If physical aggression occurred, typically both partners were aggressive. For females, exclusive engagement in physical aggression (perpetration) was reported at higher rates than exclusively being the recipient of physical aggression (victimization) and vice versa for males. Dating aggression was less prevalent among male Asian students than other ethnic groups. Engaged males and females reported the highest rates of physical aggression. Injury was reported by over 25% of males and females who reported being the recipients of physical aggression. CONCLUSIONS: Dating aggression intervention programs should address physical aggression of both males and females. Because approximately 30% of the high school males and females reported being the recipients of physical aggression by their partners, primary prevention efforts should occur before high school. PMID- 18407043 TI - Sex differences in exercise behavior during adolescence: is biological maturation a confounding factor? AB - PURPOSE: To test the hypothesis that sex-related differences in exercise behavior during adolescence are confounded by biological age. METHODS: Biological age and leisure-time exercise activity were assessed in 103 male and 83 female British Year 9 students (mean = 14.04 years, SD = 0.31). To assess exercise behavior, participants completed the Leisure-Time Exercise Questionnaire (LTEQ). Percentage of predicted adult height attained at measurement was used as an estimate of biological maturity status. Males reported more frequent engagement in strenuous forms of exercise and total exercise activity than females. RESULTS: Sex differences in exercise behavior were attenuated and nonsignificant when biological maturity was controlled for. CONCLUSIONS: Sex-related differences in biological maturity contribute to sex-related differences in exercise behavior during adolescence. PMID- 18407044 TI - Social-ecological correlates of active commuting to school among high school students. AB - PURPOSE: It has been suggested that health benefits from physical activity may be accrued through active commuting to school. Considering that active commuting is modifiable via policy and that there is limited research examining active commuting among high school students, this is a domain that warrants further investigation. The purpose of this study is to explore the relationships between demographic, behavioral, social/psychological, and environmental correlates of active commuting to school among a sample of high school students. METHODS: Students (N = 21,345) from 76 Ontario high schools (grades 9-12) completed the School Health Action, Planning and Evaluation System Physical Activity Module between 2005-2006. Demographic (gender, grade, body mass index), behavioral (smoking status, physical activity, sedentary time), social/psychological (perceived athletic ability, weight status, parental encouragement), and environmental (school location, type, weather) predictors of active commuting (daily or mixed mode) were assessed. RESULTS: Only 42.5% of high school students reported actively commuting to school. Students were less likely to actively commute to school if they were girls, in grade 12, smoked daily, were low moderate in physical activity, or attended a rural school. CONCLUSIONS: Patterns of active commuting to school are influenced by multiple factors, some of which are modifiable through intervention. This has important implications for future school-based programming designed to enhance health and physical activity of adolescents through using active modes of transportation to school. PMID- 18407045 TI - Parenting practices, parents' underestimation of daughters' risks, and alcohol and sexual behaviors of urban girls. AB - PURPOSE: In urban economically distressed communities, high rates of early sexual initiation combined with alcohol use place adolescent girls at risk for myriad negative health consequences. This article reports on the extent to which parents of young teens underestimate both the risks their daughters are exposed to and the considerable influence that they have over their children's decisions and behaviors. METHODS: Surveys were conducted with more than 700 sixth-grade girls and their parents, recruited from seven New York City schools serving low-income families. Bivariate and multivariate analyses examined relationships among parents' practices and perceptions of daughters' risks, girls' reports of parenting, and outcomes of girls' alcohol use, media and peer conduct, and heterosexual romantic and social behaviors that typically precede sexual intercourse. RESULTS: Although only four parents thought that their daughters had used alcohol, 22% of the daughters reported drinking in the past year. Approximately 5% of parents thought that daughters had hugged and kissed a boy for a long time or had "hung out" with older boys, whereas 38% of girls reported these behaviors. Parents' underestimation of risk was correlated with lower reports of positive parenting practices by daughters. In multivariate analyses, girls' reports of parental oversight, rules, and disapproval of risk are associated with all three behavioral outcomes. Adult reports of parenting practices are associated with girls' conduct and heterosexual behaviors, but not with their alcohol use. CONCLUSION: Creating greater awareness of the early onset of risk behaviors among urban adolescent girls is important for fostering positive parenting practices, which in turn may help parents to support their daughters' healthier choices. PMID- 18407046 TI - Effect of snacking frequency on adolescents' dietary intakes and meeting national recommendations. AB - PURPOSE: To determine how snacking level impacts intake of nutrients and food groups and assists in meeting recommendations outlined in the U.S. Department of Agriculture's MyPyramid Food Guidance System. METHODS: Dietary data based on 24 hour recall from 4357 adolescents 12-19 years of age participating in the National Health and Nutrition Examination Survey 2001-2004 (NHANES) were analyzed. Regression analyses were applied to examine the effect of snacking on nutrient and food group intake and to determine its effect on the likelihood of meeting MyPyramid recommendations. RESULTS: Food energy, carbohydrate, total sugars, and vitamin C intake were positively associated, whereas protein and fat intake were negatively associated, with snacking frequency. Fruit intake increased, whereas solid fat intake decreased, as snacking incidence rose. Increasing snacking frequency was also associated with a greater likelihood of meeting milk and oil recommendations for boys and meeting fruit recommendations for both genders. Non-Hispanic black adolescents were less likely to meet their milk recommendations at low and high snacking levels and more likely to meet their fruit recommendations at high levels only. Foods consumed as snacks provided 12-39% of the day's total number of portions of the five MyPyramid food groups, 35% of total discretionary calorie intake, and 43% of total added sugar intake. CONCLUSIONS: Snacking frequency affects intake of macronutrients and a few micronutrients and promotes consumption of fruits. Top food choices for snacks provide an excess of discretionary calories in the form of added sugars and fats. Modification of these choices would assist adolescents in consuming diets more consistent with national recommendations. PMID- 18407047 TI - The association between body mass index in adolescence and obesity in adulthood. AB - PURPOSE: This study used data from the National Longitudinal Study of Youth 1979 to examine the association between body mass index (BMI) in adolescence and obesity in adulthood. METHODS: Measurements of height and weight from 1981 and 2002 were used to calculate BMI for a cohort of 1309 adolescents at baseline and during adulthood. Associations between BMI at age 16/17 and obesity (BMI > or =30) at age 37/38 were analyzed using logistic regression analysis. RESULTS: When the predicted probability of adult obesity equaled 0.5, the point on the adolescent BMI distribution was close to the 85th percentile for both sexes (83rd percentile for females and 86th percentile for males). Among adolescents with a BMI in the 85th-<95th percentile, 62% of the males and 73% of the females became obese adults. Among those with a BMI > or =95th percentile, 80% of the males and 92% of the females became obese adults. Versus those with a BMI <85th percentile, those with a BMI in the 85th-<95th percentile were more likely to be obese (odds ratio = 7 for males, 11 for females) as adults, and those with a BMI > or =95th percentile were most likely to be obese (odds ratio = 18 for males, 49 for females) as adults. CONCLUSION: Adolescents with a BMI > or =85th percentile are at elevated risk for obesity in adulthood. To prevent the development of obesity and its associated health risks, population-based efforts combined with targeted interventions for these high-risk adolescents are needed. PMID- 18407048 TI - Innovations in preventive mental health care services for adolescents. AB - PURPOSE: This descriptive study examined programs designed to provide mental health-related preventive services to at-risk adolescents. METHODS: Qualitative interviews were conducted with state and local program directors and key personnel in public health and mental health departments and academic researchers who have initiated preventive services. RESULTS: States and local communities offer varied mental health-related preventive services for high-risk adolescents in diverse settings. Services include public education, screening, early intervention for adolescents, and educational programs for primary care providers. Funding mechanisms include state general funds, foundation grants, and Medicaid and the State Children's Health Insurance Program dollars. Evaluation is essential to maintain services and to persuade funders and other stakeholders to sustain these efforts. CONCLUSIONS: State and local prevention-related mental health programs have been implemented with limited funding, but significant local advocacy and community support. More extensive evaluation and cost effectiveness studies may encourage policymakers to expand services. Further data are necessary to determine how prevention-related mental health programs can best serve vulnerable youth. PMID- 18407049 TI - Waterpipe tobacco smoking on a U.S. College campus: prevalence and correlates. AB - PURPOSE: Waterpipe tobacco smoking is reported to be growing in popularity, particularly among college students. This study examined the prevalence of waterpipe tobacco smoking prevalence and perceptions in a university-based population. METHOD: This was a cross-sectional Internet-based survey of first year university students, which examined waterpipe tobacco smoking and other tobacco use, risk perceptions, influences, and perceived social acceptability. RESULTS: Waterpipe tobacco smoking within the past 30 days was reported by 20% (151/744). Relative to never users, users were more likely to perceive waterpipe tobacco smoking as less harmful than cigarette use. CONCLUSIONS: Because waterpipe tobacco smoking is increasing in prevalence and because it can involve toxicant inhalation at even greater levels than with cigarette smoking, it represents a growing public health issue. PMID- 18407050 TI - Mycobacterium fortuitum and anaerobic breast abscess following nipple piercing: case presentation and review of the literature. AB - PURPOSE: Body piercing has become increasingly prevalent. We describe a case of breast infection with combined mycobacteria and anaerobe following nipple piercing, and review the literature. CASE: A 17-year-old female developed a breast abscess 4 months after nipple piercing. Cultures grew Prevotalla melangenica and Mycobacterium fortuitum. She required drainage and antibiotic treatment. Three months into her treatment she stopped her medications, relapsed, and required drainage. Two months later, on antimycobacteria therapy, her wound is healing. DISCUSSION: Review of the infectious complications of nipple piercing yielded 12 cases, 5 of which had a foreign body. The pathogens isolated (coagulase negative staphylococcus, mycobacteria, streptococcus, anaerobe, and gordonia) are not the usual organisms to be isolated from a breast abscess. This could result from reporting bias or the presence of a foreign body, the nipple ring. The three cases of mycobacteria, in addition to ours, are reviewed. The average age is 22 years. Three to 9 months elapsed between piercing and infection. All cases required drainage. Antimycobacteria therapy was used in three of the four cases for 10 days to 6 months. CONCLUSION: With the increasing prevalence of body piercing, it is important to document and report infections. We describe a breast abscess following nipple piercing with combined anaerobic and a mycobacterial pathogens. This underscores the need for obtaining cultures including anaerobes and mycobacteria. PMID- 18407051 TI - Specialization in musculoskeletal physiotherapy--the Australian model. PMID- 18407053 TI - The R1947X mutation of NF1 causing autosomal dominant neurofibromatosis type 1 in a Chinese family. AB - Neurofibromatosis type 1 is a common autosomal dominant disorder with a high rate of penetrance. It is caused by the mutation of the tumor suppressor gene NF1, which encodes neurofibromin. The main function of neurofibromin is down regulating the biological activity of the proto-oncoprotein Ras by acting as a Ras-specific GTPase activating protein. In this study, we identified a Chinese family affected with neurofibromatosis type 1. The known gene NF1 associated with NF1 was studied by linkage analysis and by direct sequencing of the entire coding region and exon-intron boundaries of the NF1 gene. The R1947X mutation of NF1 was identified, which was co-segregated with affected individuals in the Chinese family, but not present in unaffected family members. This is the first report, which states that the R1947X mutation of NF1 may be one of reasons for neurofibromatosis type 1 in Chinese population. PMID- 18407052 TI - Mitochondrial retrograde regulation tuning fork in nuclear genes expressions of higher plants. AB - In plant cells, there are three organelles: the nucleus, chloroplast, and mitochondria that store genetic information. The nucleus possesses the majority of genetic information and controls most aspects of organelles gene expression, growth, and development. In return, organelles also send signals back to regulate nuclear gene expression, a process defined as retrograde regulation. The best studies of organelles to nucleus retrograde regulation exist in plant chloroplast to-nuclear regulation and yeast mitochondria-to-nuclear regulation. In this review, we summarize the recent understanding of mitochondrial retrograde regulation in higher plant, which involves multiple potential signaling pathway in relation to cytoplasmic male-sterility, biotic stress, and abiotic stress. With respect to mitochondrial retrograde regulation signal pathways involved in cytoplasmic male-sterility, we consider that nuclear transcriptional factor genes are the targeted genes regulated by mitochondria to determine the abnormal reproductive development, and the MAPK signaling pathway may be involved in this regulation in Brassica juncea. When plants suffer biotic and abiotic stress, plant cells will initiate cell death or other events directed toward recovering from stress. During this process, we propose that mitochondria may determine how plant cell responds to a given stress through retrograde regulation. Meanwhile, several transducer molecules have also been discussed here. In particular, the Paepe research group reported that leaf mitochondrial modulated whole cell redox homeostasis, set antioxidant capacity, and determined stress resistance through altered signaling and diurnal regulation, which is an indication of plant mitochondria with more active function than ever. PMID- 18407054 TI - Cloning and characterization of nanos gene in silkworm Bombyx mori. AB - Gene nanos is a maternal posterior group gene required for normal development of abdominal segments and the germ line in Drosophila. Expression of nanos-related genes is associated with the germ line in a broad variety of other taxa. In this study, the 5'-RACE method and the in silico cloning method are used to isolate the new nanos-like gene of Bombyx mori and the gene obtained is analyzed with bioinformatics tools. The putative protein is expressed in Escherichia coli and the antiserum has been produced in New Zealand white rabbits. The result shows that the nanos cDNA is 1,913 bp in full length and contains a 954 bp open reading frame. The deduced protein has 317 amino acid residues, with a predicted molecular weight of 35 kDa, isoelectric point of 5. 38, and contains a conserved nanos RNA binding domain. The conserved region of the deduced protein shares 73% homology with the nanos protein conserved region of Honeybee (Apis mellifera). This gene has been registered in the GenBank under the accession number EF647589. One encoding sequence of the nanos fragment has been successfully expressed in E. coli. Western blotting analysis indicates that homemade antiserum can specifically detect nanos protein expressed in prokaryotic cells. PMID- 18407055 TI - Mutation analyses of integrated HBV genome in hepatitis B patients. AB - Little has been learnt in the last 30 years about detection of HBV genome as well as its mutation analysis between hepatitis B fathers (HBF) and their children. In this study, we used nest polymerase chain reaction (PCR), fluorescence in situ hybridization (FISH), and DNA sequencing analysis, to examine the integrated HBV genome in paraffin-embedded testis tissues, which were taken as samples from HBF, and in peripheral blood mononuclear cells (PBMC) from 74 cases of HBFs and their children who were born after their fathers' HBV infection (caHBF). We found that HBV DNA existed in testis tissues, mainly in the basilar parts of the seminiferous tubules, and also in PBMC of HBF. It was also documented that there were point mutations of poly-loci, insertions and deletions of nucleotides in integrated HBV genomes, and the types of gene mutations in the HBFs were similar to those in caHBF. This study addresses the major types of gene mutations in integrated HBV genome in human patients and also presents reliable evidence of possible genetic transmission of hepatitis B. PMID- 18407056 TI - Association of polymorphisms of Nramp1 gene with immune function and production performance of Large White pig. AB - The present research was designed to study the association of polymorphism of natural resistance-associated macrophage protein1 (Nramp1) with some immune function and the production performance in Large White pig. The PCR-RFLP technique was applied to analyze the correlation between the polymorphisms of Nramp1 gene and immune function [value of Polymorphonuclear Leukocytes (PMN) obtained by Nitroblue Tetrazolium (NBT) Reduction and effect of Cytotoxin in Monocyte] and production performance in 165 Large White pigs. The results showed that there was one Nde I restriction locus in Large White pig, and both values of PMN by NBT Reduction and effect of Cytotoxin in Monocyte in genotype BB were higher than those in genotype AB (P<0.05). Simultaneously, the weight of 180-day old pigs with genotype BB was higher than that with genotype AB (P<0.05). The results indicated that there was a significant correlation between different genotypes of Nramp1 gene and immune function and production performance, and it can be regarded as a candidate gene of disease resistance. All these results provide valuable reference to further studies of pig disease resistance. PMID- 18407057 TI - Genome evolution trend of common carp (Cyprinus carpio L.) as revealed by the analysis of microsatellite loci in a gynogentic family. AB - Genome evolution arises from two main ways of duplication and reduction. Fish specific genome duplication (FSGD) may have occurred before the radiation of the teleosts. Common carp (Cyprinus carpio L.) has been considered to be a tetraploid species, because of its chromosome numbers (2n=100) and its high DNA content. Using 69 microsatellite primer pairs, the variations were studied to better understand the genome evolution (genome duplication and diploidization) of common carp from a gynogenetic family. About 48% of primer pairs were estimated to amplify duplicates based on the number of PCR amplification per individual. Segregation patterns in the family suggested a partially duplicated genome structure and disomic inheritance. This indicates that the common carp is tetraploid and polyploidy occurred by allotetraploidy. Two primer pairs (HLJ021 and HLJ332) were estimated to amplify reduction based on the number of PCR amplification per individual. One allele in HLJ002 locus and HLJ332 locus was clearly lost in the gynogenetic family and the same as in six wild populations. Segregation patterns in the family suggested a partially diplodization genome structure. A hypothesis transition (dynamic) and equilibrium (static) were proposed to explain the common carp genome evolution between genome duplication and diploidization. PMID- 18407058 TI - Genome-wide analysis of heat shock transcription factor families in rice and Arabidopsis. AB - The heat shock transcription factors (HSFs) are the major heat shock factors regulating the heat stress response. They participate in regulating the expression of heat shock proteins (HSPs), which are critical in the protection against stress damage and many other important biological processes. Study of the HSF gene family is important for understanding the mechanism by which plants respond to stress. The completed genome sequences of rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana) constitute a valuable resource for comparative genomic analysis, as they are representatives of the two major evolutionary lineages within the angiosperms: the monocotyledons and the dicotyledons. The identification of phylogenetic relationships among HSF proteins in these species is a fundamental step to unravel the functionality of new and yet uncharacterized genes belonging to this family. In this study, the full complement of HSF genes in rice and Arabidopsis has probably been identified through the genome-wide scan. Phylogenetic analyses resulted in the identification of three major clusters of orthologous genes that contain members belonging to both species, which must have been represented in their common ancestor before the taxonomic splitting of the angiosperms. Further analysis of the phylogenetic tree reveals a possible dicot specific gene group. We also identified nine pairs of paralogs, as evidence for studies on the evolution history of rice HSF family and rice genome evolution. Expression data analysis indicates that HSF proteins are widely expressed in plants. These results provide a solid base for future functional genomic studies of the HSF gene family in rice and Arabidopsis. PMID- 18407059 TI - Mapping QTLs with epistatic effects and QTL x environment interactions for plant height using a doubled haploid population in cultivated wheat. AB - Quantitative trait loci (QTLs) for plant height in wheat (Triticum aestivum L.) were studied using a set of 168 doubled haploid (DH) lines, which were derived from the cross Huapei 3/Yumai 57. A genetic linkage map was constructed using 283 SSR and 22 EST-SSR markers. The DH population and the parents were evaluated for wheat plant height in 2005 and 2006 in Tai'an and 2006 in Suzhou. QTL analyses were performed using the software of QTLNetwork version 2.0 based on the mixed linear model. Four additive QTLs and five pairs of epistatic effects were detected, which were distributed on chromosomes 3A, 4B, 4D, 5A, 6A, 7B, and 7D. Among them, three additive QTLs and three pairs of epistatic QTLs showed QTL x environment interactions (QEs). Two major QTLs, Qph4B and Qph4D, which accounted for 14.51% and 20.22% of the phenotypic variation, were located similar to the reported locations of the dwarfing genes Rht1 and Rht2, respectively. The Qph3A-2 with additive effect was not reported in previous linkage mapping studies. The total QTL effects detected for the plant height explained 85.04% of the phenotypic variation, with additive effects 46.07%, epistatic effects 19.89%, and QEs 19.09%. The results showed that both additive effects and epistatic effects were important genetic bases of wheat plant height, which were subjected to environmental modifications, and caused dramatic changes in phenotypic effects. The information obtained in this study will be useful for manipulating the QTLs for wheat plant height by molecular marker-assisted selection (MAS). PMID- 18407061 TI - Linking inflammasome activation and phagosome maturation. AB - One-third of the world's population is infected with Mycobacterium tuberculosis, one of the most effective human pathogens, whose success is attributed to the deployment of remarkably sophisticated immune evasion mechanisms. In this issue of Cell Host & Microbe, a new study unravels a novel strategy of immune evasion and enhanced bacterial intracellular survival, which is dependent on inhibition of inflammasome activation by an M. tuberculosis-encoded metalloprotease. PMID- 18407062 TI - (C)Re-combining textbook models of virus spread within the host. AB - Classic viral pathogenesis models postulate that tissues supporting efficient virus replication promote virus dissemination, which culminates in clinical illness. In this issue of Cell Host & Microbe, Sacher and colleagues use Cre/loxP recombination to label murine cytomegalovirus during replication in distinct cell types in vivo. Strikingly, they demonstrate that the most productive cell type in the host-the hepatocyte-contributes no progeny to dissemination to other tissues. PMID- 18407063 TI - Identifying host targets for drug development with knowledge from genome-wide studies: lessons from HIV-AIDS. AB - Advances in human genomics are now being effectively applied to the search for host factors underlying susceptibility to common diseases. From the steady stream of studies showing association of host genetic factors with viral diseases, it has become clear that host factors contribute substantially to the variability of viral infections in humans. Candidate gene studies that seek to show associations between single-nucleotide polymorphisms (SNPs) with a disease outcome have predominated, but whole-genome association studies (GWAS) have recently appeared. A major goal of these studies is to understand how human genetic variation contributes to individual differences in susceptibility and to exploit this knowledge for targeted drug development. PMID- 18407064 TI - P bodies, stress granules, and viral life cycles. AB - Eukaryotic mRNAs are in a dynamic equilibrium between different subcellular locations. Translating mRNAs can be found in polysomes, mRNAs stalled in translation initiation accumulate in stress granules and mRNAs targeted for degradation or translation repression can accumulate in P bodies. Partitioning of mRNAs between polysomes, stress granules, and P bodies affects rates of translation and mRNA degradation. Host proteins within P bodies and stress granules can enhance or limit viral infection, and some viral RNAs and proteins accumulate in P bodies and/or stress granules. Thus, an important interplay among P bodies, stress granules, and viral life cycles is beginning to emerge. PMID- 18407066 TI - Mycobacterium tuberculosis prevents inflammasome activation. AB - Mycobacterium tuberculosis (Mtb) parasitizes host macrophages and subverts host innate and adaptive immunity. Several cytokines elicited by Mtb are mediators of mycobacterial clearance or are involved in tuberculosis pathology. Surprisingly, interleukin-1beta (IL-1beta), a major proinflammatory cytokine, has not been implicated in host-Mtb interactions. IL-1beta is activated by processing upon assembly of the inflammasome, a specialized inflammatory caspase-activating protein complex. Here, we show that Mtb prevents inflammasome activation and IL 1beta processing. An Mtb gene, zmp1, which encodes a putative Zn(2+) metalloprotease, is required for this process. Infection of macrophages with zmp1 deleted Mtb triggered activation of the inflammasome, resulting in increased IL 1beta secretion, enhanced maturation of Mtb containing phagosomes, improved mycobacterial clearance by macrophages, and lower bacterial burden in the lungs of aerosol-infected mice. Thus, we uncovered a previously masked role for IL 1beta in the control of Mtb and a mycobacterial system that prevents inflammasome and, therefore, IL-1beta activation. PMID- 18407067 TI - Salmonella AvrA Coordinates Suppression of Host Immune and Apoptotic Defenses via JNK Pathway Blockade. AB - Salmonellae are bacterial pathogens that have evolved sophisticated strategies to evade host immune defenses. These strategies include the secretion of effector proteins into mammalian cells so as to subvert innate immune and apoptotic signaling pathways, thereby allowing Salmonella to avoid elimination. Here, we show that the secreted Salmonella typhimurium effector protein AvrA possesses acetyltransferase activity toward specific mitogen-activated protein kinase kinases (MAPKKs) and potently inhibits c-Jun N-terminal kinase (JNK) and NF kappaB signaling pathways in both transgenic Drosophila and murine models. Furthermore, we show that AvrA dampens the proapoptotic innate immune response to Salmonella at the mouse intestinal mucosa. This activity is consistent with the natural history of Salmonella in mammalian hosts, where the bacteria elicit transient inflammation but do not destroy epithelial cells. Our findings suggest that targeting JNK signaling to dampen apoptosis may be a conserved strategy for intracellular pathogens. PMID- 18407069 TI - The major virus-producing cell type during murine cytomegalovirus infection, the hepatocyte, is not the source of virus dissemination in the host. AB - The course of systemic viral infections is determined by the virus productivity of infected cell types and the efficiency of virus dissemination throughout the host. Here, we used a cell-type-specific virus labeling system to quantitatively track virus progeny during murine cytomegalovirus infection. We infected mice that expressed Cre recombinase selectively in vascular endothelial cells or hepatocytes with a murine cytomegalovirus for which Cre-mediated recombination would generate a fluorescently labeled virus. We showed that endothelial cells and hepatocytes produced virus after direct infection. However, in the liver, the main contributor to viral load in the mouse, most viruses were produced by directly infected hepatocytes. Remarkably, although virus produced in hepatocytes spread to hepatic endothelial cells (and vice versa), there was no significant spread from the liver to other organs. Thus, the cell type producing the most viruses was not necessarily the one responsible for virus dissemination within the host. PMID- 18407070 TI - Hyponatremia: endocrinologic causes and consequences of therapy. AB - Hyponatremia is a relatively common medical disorder frequently of endocrinologic origin. Although the neurologic signs and symptoms associated with hyponatremia are well known, recent clinical and experimental studies have shown that rapid correction of hyponatremia can in some cases cause a brain-demyelinating disease that can lead to consequences just as severe as those of hyponatremia itself. Understanding the physiologic mechanisms by which the brain adapts to hypoosmolar conditions has led to a better appreciation of the pathogenesis of neurologic dysfunction during both hyponatremia and its therapy, and this in turn has allowed the establishment of rational guidelines for safe therapy of hyponatremic patients. In cases such as this, however, in which a spontaneous rapid water diuresis occurs following a change in hormonal therapy, the rate and magnitude of the increase in serum [Na+J may exceed these guidelines unless active intervention is undertaken to interrupt the induced diuresis. PMID- 18407071 TI - Changes in thyroid function in nonthyroid illness. AB - Nonthyroid illness can cause changes in thyroid function that have been described as low triiodothyronine (T(3)) and low thyroxine (T(4)) states. Reduced peripheral conversion of T4 to T(3) explains the low serum T(3) concentration. The explanation for the low serum T(4) level is multifactorial; whether free thyroxine (FT(4)) level is normal or reduced remains controversial. Cytokines such as tumor necrosis factor alpha, which are produced by the immune system during severe illness, may inhibit thyroid function directly and be responsible for the changes in pituitary-thyroid function. PMID- 18407072 TI - Control of thyroid cell proliferation and goitrogenesis. AB - Human thyroid cells proliferate during development and in adults in response to physiologic and pathologic stimuli. Under normal conditions, they turn over about once every 8 years. The main physiologic regulators are thyrotropin and iodide and, in disease, thyroid-stimulating and thyroid-blocking antibodies. Growth factors modulate proliferation in vitro, but their role in vivo is still unknown. Mitogenic effects are mediated via three major pathways: the cyclic AMP, protein tyrosine kinase, and the Ca(2+) phosphatidylinositol cascades. In this review, the role of these cascades in hyperthyroidism, congenital thyroid defects, and autonomous adenoma is analyzed. PMID- 18407068 TI - Human cytomegalovirus protein UL38 inhibits host cell stress responses by antagonizing the tuberous sclerosis protein complex. AB - Human cytomegalovirus proteins alter host cells to favor virus replication. These viral proteins include pUL38, which prevents apoptosis. To characterize the mode of action of pUL38, we modified the viral genome to encode an epitope-tagged pUL38 and used rapid immunoaffinity purification to isolate pUL38-interacting host proteins, which were then identified by mass spectrometry. One of the cellular proteins identified was TSC2, a constituent of the tuberous sclerosis tumor suppressor protein complex (TSC1/2). TSC1/2 integrates stress signals and regulates the mammalian target of rapamycin complex 1 (mTORC1), a protein complex that responds to stress by limiting protein synthesis and cell growth. We showed that pUL38 interacts with TSC1 and TSC2 in cells infected with wild-type cytomegalovirus. Furthermore, TSC1/2 failed to regulate mTORC1 in cells expressing pUL38, and these cells exhibited the enlarged size characteristic of cytomegalovirus infection. Thus, pUL38 supports virus replication at least in part by blocking cellular responses to stress. PMID- 18407074 TI - Progress and approaches in mapping the surfaces of human follicle-stimulating hormone: comparison with the other human pituitary glycoprotein hormones. AB - The pituitary glycoprotein hormones, including human follicle-stimulating hormone (hFSH), are involved in the physiologic functions of receptor binding, in subunit association during assembly and processing, and in untoward effects such as antibody formation during clinical intervention. These activities derive from specific macromolecular associations; a major research trend has been to map the surfaces of these glycoprotein hormones (hFSH, hLH, hCG, and hTSH) into discrete areas responsible for each activity by using a variety of biochemical approaches. Each surface domain or determinant of the hormone is likely to comprise discontinuous amino acid sequences, from one or both subunits, assembled into a precise, unique, macromolecular surface. The paradigm of antigen-antibody interaction may help to explain how the surfaces are assembled, how the common alpha-subunit combines with the unique beta-subunit of each, and how the receptor interacts with heterodimeric hormone. PMID- 18407073 TI - Thymocyte apoptosis: a model of programmed cell death. AB - Recently there has been widespread appreciation for the role of apoptosis, or programmed cell death, in the maintenance of tissue structure and function. Studies in several model systems have revealed that apoptosis is profoundly regulated by a number of diverse hormones, including steroids. The killing of immature thymic lymphocytes by glucocorticoids has emerged as an important model to define the biochemical mechanisms that mediate the programmed cell death process. Using this model, we, and others, have shown that lymphocytes degrade their DNA in response to glucocorticoids. The onset of DNA degradation precedes cell death and is the probable cause of apoptosis. This unique response to endocrine signal transduction will undoubtedly promise new insights into the mechanism of hormone action. PMID- 18407065 TI - Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. AB - We have investigated the interrelationship between diet, gut microbial ecology, and energy balance using a mouse model of obesity produced by consumption of a prototypic Western diet. Diet-induced obesity (DIO) produced a bloom in a single uncultured clade within the Mollicutes class of the Firmicutes, which was diminished by subsequent dietary manipulations that limit weight gain. Microbiota transplantation from mice with DIO to lean germ-free recipients promoted greater fat deposition than transplants from lean donors. Metagenomic and biochemical analysis of the gut microbiome together with sequencing and metabolic reconstructions of a related human gut-associated Mollicute (Eubacterium dolichum) revealed features that may provide a competitive advantage to members of the bloom in the Western diet nutrient milieu, including import and processing of simple sugars. Our study illustrates how combining comparative metagenomics with gnotobiotic mouse models and specific dietary manipulations can disclose the niches of previously uncharacterized members of the gut microbiota. PMID- 18407075 TI - Clinical applications of GnRH and its analogues. AB - The chemical characterization of the hypothalamic decapeptide gonadotropin releasing hormone (GnRH) has stimulated the development of analogues of GnRH with important clinical applications. Chronic administration of the GnRH analogues nafarelin acetate and leuprolide acetate results in an initial stimulation of pituitary luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion, which is followed by a profound decrease in the secretion of LH and FSH. The decrease in the secretion of LH and FSH produces a hypogonadal state that is associated with an improvement in many sex-steroid-dependent disease processes. The GnRH analogues are clearly effective in the treatment of prostate cancer, endometriosis, uterine myomas, polycystic ovarian disease, and the premenstrual syndrome. PMID- 18407076 TI - Glycoprotein hormone alpha-subunit in pituitary adenomas. AB - Elevated serum glycoprotein hormone alpha-subunit (alpha-subunit) levels are seen in about one of six patients bearing pituitary adenomas. This finding has particular clinical significance in patients with nonfunctioning pituitary adenomas. Moreover, the measurement of alpha-subunit along with the calculation of the molar ratio between alpha-subunit and TSH, LH, or FSH is helpful in the diagnosis of glycoprotein hormone-secreting pituitary adenomas. Since serum alpha subunit levels may vary greatly in several physiologic and pathologic conditions, care has to be taken to differentiate abnormal from normal states of alpha subunit hypersecretion as well as to exclude causes of alpha-subunit overproduction only casually associated with the presence of pituitary tumors. PMID- 18407077 TI - Growth hormone therapy in adults. AB - The benefits of growth hormone (GH) replacement therapy of adults are currently being evaluated, and initial results indicate that 4 or 6 months of GH therapy alter body composition, with an increase in muscle and a decrease in adipose tissue mass. It remains to be determined whether this treatment substantially improves function. PMID- 18407078 TI - Molecular basis of inherited thyroxine-binding globulin defects. AB - Thyroxine-binding globulin (TBG) is a liver glycoprotein that transports thyroid hormones in serum. Inherited TBG defects appear as partial or complete deficiency and TBG excess. Sequencing of the TBG gene located on the X-chromosome has revealed nucleotide substitutions in partial TBG deficiency, and substitutions or deletions in complete deficiency variants. Whereas the deduced changes of the primary structure of the protein have been sufficient to explain the observed alterations of properties in some of the TBG variants, this has not been the case in other inherited TBG defects studied at the gene level. Further analysis of these and other variants may provide helpful information on glycoprotein synthesis and processing and on protein-hormone interaction. PMID- 18407079 TI - Different forms of the prolactin receptor: insights into the mechanism of prolactin action. AB - The primary structure of two forms (short [approximately 300 amino acids] and long [approximately 600 amino acids]) of prolactin (PRL) receptor have been deduced by cloning their cDNAs. PRL receptors belong to the superfamily of cytokine growth hormone-PRL receptors, based on conserved sequences in their extracellular domains. No clear second messenger of PRL has been identified, and because of the wide range of actions associated with PRL, it is doubtful that a single unifying mechanism will be found. Mutagenesis of extracellular and intracellular domains of the PRL receptor should help define subdomains involved in hormone binding, and coupled with a functional assay, delineate regions necessary for signal transduction. PMID- 18407080 TI - Ontogeny of thyrotropin-releasing hormone biosynthesis and release in hypothalamic neurons. AB - Thyrotropin-releasing hormone (TRH) is expressed at early postmitotic stages of hypothalamic neuron development, in the mouse and rat, as revealed by the presence of the mature peptide, of pro-TRH mRNAs, and of large precursor forms. This indicates a coordinate expression of several genes encoding, respectively, pro-TRH, its processing enzymes, and the cell machinery for intracellular transport, sorting, and release of TRH. During development, an acceleration of pro-TRH processing is revealed by an increased proportion of the mature peptide. This is correlated with changes in the respective distribution of pro-TRH and TRH along neurites and the ontogenesis of neurosecretory granules. PMID- 18407081 TI - Follistatin/activin-binding protein. AB - Follistatin, or follicle-stimulating hormone (FSH) suppressing protein, is a protein isolated from side fractions in the purification of inhibin and activin, which shares with inhibin the property of suppressing the secretion of FSH from the pituitary gonadotroph. This protein is structurally distinct from inhibin. Recent observations that follistatin is an activin-binding protein suggest that it may have a much wider biologic role, given the variety of systems in which activin has been shown to act. PMID- 18407082 TI - Complete androgen insensitivity Pathophysiology, diagnosis, and management. AB - The syndrome of complete androgen insensitivity is an X-linked inherited disorder resulting in marked inhibition of androgen action. The following case illustrates a subject with complete androgen insensitivity who, despite being a genetic and gonadal male, presents as a phenotypic female with primary amenorrhea, normal breast development, and lack of axillary and pubic hair. The diagnosis, pathophysiology, and management of the condition are discussed, as well as recently identified abnormalities in the androgen-receptor gene. The partial forms of androgen insensitivity are also included in the discussion. PMID- 18407083 TI - Calcitonin peptide inhibition of TRH-stimulated prolactin secretion Additional evidence for inhibitory regulation of phospholipase C. AB - Calcitonin peptides have been reported to exert direct inhibitory effects on stimulated prolactin secretion from lactotrophs. Several studies indicate that calcitonin peptide inhibition is rather selective for the stimulatory effects of thyrotropin-releasing hormone (TRH), but not those of other secretagogues. Recent reports demonstrate inhibitory effects of calcitonin peptides on TRH-induced calcium mobilization and inositol phosphate generation. The possibility is discussed that calcitonin peptides act at pituitary receptors that are coupled to phospholipase C in an inhibitory manner. PMID- 18407084 TI - Natriuretic peptides A family of hormones. AB - The natriuretic peptides are a family of structurally related peptides, products of at least three genes, which share a common, 17-membered, internal disulfide ring. Three major subtypes (A-, B-, and C-type natriuretic peptides) exist, and each is present in various tissues in multiple N- and C-terminally extended or shortened forms. The recent description of the C-type peptide CNP, the major brain-derived peptide, has suggested that original studies examining the endocrine actions of the natriuretic peptides using A-type (ANP) or B-type (BNP) isoforms might have underestimated some of the biopotency of the natriuretic peptides or even failed to detect the full spectrum of the family's bioactivity. The identification of multiple, often species-specific, subtypes of the natriuretic peptides, together with the characterization of at least two classes of receptors for these hormones, indicates a variety of potential avenues for the development of therapeutic strategies for the use of these factors. Importantly, the description of protease inhibitors that prolong and enhance the actions of the natriuretic peptides in humans suggests potential for the recruitment of the natriuretic peptides in the treatment of endocrine as well as cardiovascular disease. PMID- 18407085 TI - The physiology of decidual prolactin and other decidual protein hormones. AB - Human decidual tissue, the specialized endometrium of the luteal phase of the menstrual cycle and pregnancy, synthesizes and releases prolactin, relaxin, renin, and at least three insulinlike growth factor (IGF)-binding proteins. Although the physiologic roles of these decidual protein hormones during pregnancy are unclear, numerous studies suggest that the hormones may act locally to affect the function of the placenta, decidua, and fetal membranes. In addition, the synthesis and release of these hormones appear to be regulated locally by factors produced by the placenta, decidua, and fetal membranes. PMID- 18407086 TI - The role of thyroid hormone in amphibian metamorphosis. AB - Recent advances in our understanding of thyroid hormone (TH) metabolism, and the nature and function of TH receptors, have shed some light on the mechanisms by which TH initiates the numerous morphologic and biochemical components of amphibian metamorphosis. The data support the view that the successful accomplishment of metamorphosis depends primarily on a series of programmed increases in the rate of secretion of TH. In addition, as climax approaches and the requirement for TH increases, the physiologic effectiveness of the circulating hormone is enhanced as a result of changes in peripheral TH economy and tissue-specific increases in TH receptor concentration. The receptor bears a striking resemblance to its mammalian and avian counterparts, and it is postulated that TH-induced metamorphic events result, at least in part, from changes in transcriptional activity induced by the effect of triiodothyronine receptor complexes on the response element of the relevant genes. PMID- 18407087 TI - Prolactin variants. AB - Prolactin (PRL) is one of the most versatile hormones of the pituitary in terms of biologic actions, but how a single molecule is able to evoke so many different responses in the organism is not known. Research in recent years has uncovered a surprising degree of structural polymorphism for PRL, the different forms having variable biopotencies. Such findings lend credence to the hypothesis that the molecular heterogeneity of PRL is one of the mechanisms for creating diversity in the biologic actions of this hormone. PMID- 18407088 TI - The thyroid and osteoporosis. AB - Increasingly sophisticated tests of thyroid function have indicated that minor degrees of hyperthyroidism are common, especially in patients taking thyroxine (T(4)) therapy. Several recent reports have suggested that such patients have reduced bone density as a consequence of their hyperthyroidism. Further studies are required to determine whether these changes are found in all age groups and both sexes and whether they are clinically relevant in terms of risk of osteoporotic fractures. Meanwhile, it is prudent to avoid excessive doses of thyroid hormones for patients requiring replacement therapy. PMID- 18407089 TI - Growth Hormone What is it and what does it do? AB - The evidence is now irrefutable that growth hormone (GH), long thought to be a single substance, is actually a mixture of several different forms. These multiple forms must be a consideration in any physiologic study if an accurate evaluation of the actions of GH is to be made. PMID- 18407090 TI - Excitatory amino acid neurotransmission evidence for a role in neuroendocrine regulation. AB - There is compelling evidence that endogenous excitatory amino acid neurotransmission is an important component of the neuroendocrine transmission line that regulates anterior pituitary-hormone release and, thus, reproduction. Excitatory amino acids (EAAs), such as glutamate and aspartate, are found in large quantities in neuroendocrine tissues such as the hypothalamus, and neurons from a variety of hypothalamic nuclei respond with marked excitation to EAA application. Exogenous EAA administration rapidly increases the release of GnRH, LH, and prolactin secretion in vivo and in vitro. Antagonist studies demonstrate that EAA-receptor activation is involved in a number of reproductive-endocrine events, such as the induction of puberty, seasonal breeding, steroid-induced LH secretion, and the preovulatory surge of LH and prolactin in the female. EAA regulation of these neuroendocrine events appears to be achieved through modulation and regulation of hypothalamic GnRH secretion. PMID- 18407091 TI - Regulation of hypothalamic neuropeptide expression by peripheral metabolism. AB - The mechanisms of appetite and body-weight regulation by peripheral signals are highly complex in vertebrates and remain poorly understood. It is intuitively apparent that such regulation must involve interactions between peripheral metabolic status and the brain, but what are the signals recognized by the brain to initiate feeding? The hypothalamus has long been recognized as central in "recognition" of peripheral nutrient and metabolic signals (and, perhaps, body weight status) and in "regulation" of hunger and satiety responses and, therefore, is a logical site on which to focus research aimed at understanding interactions between and regulation of the periphery and central nervous system. Recent studies demonstrating modulation of hypothalamic neurotransmitter expression by peripheral metabolic status may yield insights into regulation of appetite and metabolism in obesity and aberrant metabolic homeostasis. This review concentrates on summarizing data regarding regulation of expression of neuropeptide Y and growth hormone-releasing hormone as model peptide systems for addressing questions relating peripheral metabolism and hypothalamic neuropeptide expression. PMID- 18407092 TI - Proprotein and prohormone convertases of the subtilisin family Recent developments and future perspectives. AB - Limited proteolysis of precursors at specific pairs of basic residues and/or at single basic amino acids is a widespread mechanism by which the cell expresses a repertoire of biologically active proteins and peptides. The cloning and cellular expression of the yeast KEX2 gene product demonstrated that this enzyme belongs to the subtilisin family of serine proteinases, and that it exhibits exquisite selectivity for cleavage post pairs of basic residues in a number of yeast and mammalian precursors. The search for the homologous mammalian convertases led to the identification and molecular cloning of three new members of the family, furin, PCI, and PC2. Whereas furin is almost ubiquitous, PCI and PC2 localize mostly in endocrine and neuroendocrine tissues and cells. Coexpression of each gene product with proproteins demonstrated that each proteinase selectively cleaved these precursors at distinct pairs of LysArg and ArgArg residues. In human and mouse, the genes coding for furin, PCI, and PC2 reside on three different chromosomes. Overexpression of PO and PC2 in Sf9 cells in the baculovirus system demonstrated that these enzymes are not secreted and that they both retained their N-terminal prodomain. PMID- 18407094 TI - Endothelin an endocrine role. AB - Endothelin is a potent and long-acting vasoconstrictor first isolated from the vascular endothelium. It was found in the search for the long-postulated endothelial vascular smooth muscle activator. Recent reports, however, suggest that endothelin may affect the release of the pituitary hormones and control the levels of atrial natriuretic peptide, renin, and the catecholamines. Its widespread distribution within the CNS and other tissues suggests that endothelin may have an important function as a neurotransmitter. PMID- 18407093 TI - The role of the immunoendocrine interaction via the hypothalamo-pituitary-adrenal axis in autoimmune disease Emphasis on the obese strain chicken model. AB - The key to the molecular basis of the pathogenesis of autoimmune disease (AID) certainly lies in the identification of genes coding for the altered immune response and those responsible for the susceptibility of the target organ to the autoimmune attack. The elucidation of nonessential modulatory factors is, however, also of great importance, because it is via these routes that an effective AID therapy is practical at the present time. Hormones in general, and those affecting the hypothalamo-pituitary-adrenal (HPA) axis in particular, are among the best candidates for more rational new therapeutic approaches. Studies of immune-endocrine communication in animal models with spontaneously occurring organ-specific or systemic autoimmune diseases are ideal for this kind of investigation, since immunologic studies commencing prior to onset of the AID in question can be performed. PMID- 18407095 TI - Insulinotropic glucagonlike peptide-I(7-37)/(7-36)amide A new incretin hormone. AB - Intestinal peptide hormones, released into circulation in response to a meal, are important augmentors of the postprandial insulin release stimulated by absorbed nutrients. In this "Incretin concept" glucagon-like peptide I(7-37) plays a major role because it is the most powerful glucose-dependent insulin secretagogue described so far. Glucagonlike peptide I(7-37) also stimulates proinsulin gene expression and proinsulin biosynthesis in insulinoma cells, and it may be involved in the regulation of the intracellular insulin pool of the B cell. Recent studies show elevated levels of glucagonlike peptide I(7-37) in patients with non-insulin-dependent diabetes mellitus. The physiology and pathophysiology of the enteroinsular axis is a promising field of basic and clinical research that has a relevance to diabetes mellitus. PMID- 18407096 TI - Molecular basis of gonadotropin receptor regulation. AB - The anterior pituitary hormones, luteinizing hormone (LH) and follicle stimulating hormone (FSH), act upon the ovary and testis via occupancy of specific cell membrane receptors, resulting in increased cAMP production, steroidogenesis, and expression of differentiation-related genes. Recent cloning of the cDNAs for LH and FSH receptors allows the analysis of mRNA levels for these receptors in gonadal tissues. This review summarizes progress in elucidating the molecular basis of LH and FSH receptor gene regulation in the ovary and testis during different physiologic states. PMID- 18407097 TI - Epidemiology of iodine deficiency. AB - Iodine deficiency (ID) causes goiter, cretinism, neonatal hypothyroidism, irreversible mental retardation, and child and infant death. Over one billion people are at risk, most of them in developing countries. While ID is the primordial factor in these conditions, other environmental and host factors significantly modify the magnitude and clinical presentation of iodine deficiency disorders. The interactions and mechanisms by which these factors operate are complex and mostly unknown, requiring more investigation. PMID- 18407098 TI - Neuroendocrine control of growth hormone secretion in humans. AB - The episodic secretion of growth hormone (GH) depends on the rhythmic alternation in the hypothalamic release of GHRH and somatostatin (SS) into the hypophyseal portal system. In turn, GH appears to maintain this rhythm by stimulating SS and inhibiting GHRH secretion. Central adrenergic pathways, by modulating SS secretion, seem to be the final mediator for most stimuli, including other neurotransmitters, modifying GH release. Glucocorticoids enhance GH gene transcription and facilitate GH gene expression; the latter effect also depends on physiologic plasma levels of thyroid hormones. Sex steroids mainly act on SS neurons, most likely by affecting the alpha(2)-adrenergic transmission to them. Metabolic intermediates can also affect GH secretion: arginine and hypoglycemia inhibit SS release, whereas hyperglycemia and free fatty acids (FFA) stimulate it. In addition, a strong inhibitory effect of FFA on the somatotrophs also occurs. PMID- 18407099 TI - Pulsatile patterns in hormone secretion. AB - Endocrine systems are regulated dynamically. With the development of sensitive methods for hormone measurements and high-frequency blood sampling, it has been shown in many endocrine systems that hormonal information is encoded in distinct pulses varying in frequency from minutes to hours. Focusing on pituitary hormones as an example, this review discusses the relevance of this pulsatile pattern of secretion on the regulation of endocrine systems and its implications on diagnosis and therapy o f endocrine diseases. PMID- 18407100 TI - Long-term treatment with RU486 and glucocorticoid receptor resistance: theoretical and therapeutic implications. AB - Glucocorticoid receptor resistance in humans occurs rarely in hereditary disorders. RU486 blocks progesterone receptors, but at higher doses also blocks glucocorticoid receptors. In this article, the clinical and biochemical effects of long-term treatment with RU486 of patients with meningiomas, breast cancer, and Cushing's syndrome are compared with those present in patients with hereditary glucocorticoid receptor resistance. The potential use of RU486 outside the field of fertility regulation is discussed. PMID- 18407101 TI - Acromegaly. AB - In the majority of cases, acromegaly is due to GH hypersecretion by a somatotroph pituitary tumor. The etiology of acromegaly is not known, and may be related to GHRH hypersecretion, intrinsic pituitary defect, or a combination thereof. Recent physiologic data and molecular biology techniques provide insights into the pathophysiology of this condition. Treatment options include surgery, radiation, and judicious administration of pharmacologic compounds inhibiting GH secretion and tumor growth. PMID- 18407102 TI - Angiotensin II receptor antagonists and receptor subtypes. AB - Recently discovered nonpeptide angiotensin II receptor antagonists represent a new class of potential drugs for the treatment of hypertension and congestive heart failure. Further, these antagonists have been successfully used as selective research tools for physiologic studies of angiotensin H and defining angiotensin II receptor subtypes. PMID- 18407103 TI - Local extragonadal roles of activins. AB - Activins were initially isolated from gonadal fluids based on their ability to stimulate FSH secretion from gonadotropes of the anterior pituitary. They represent a group of homo/heterodimeric proteins that are structurally related to the transforming growth factor beta superfamily of growth and differentiation factors. The importance of activins as autocrine and paracrine modulators of a diverse set of biologic functions is now appreciated based on their widespread anatomic distribution and numerous biologic activities. PMID- 18407104 TI - Epidemiology of osteoporosis. AB - Osteoporosis is a major public health problem through its association with age related fractures. Although fracture risk at any skeletal site depends upon a complex interaction between bone strength and trauma, recent epidemiologic studies confirm that bone density is currently the best single predictor of future fracture. The increasing burden of osteoporotic fractures urgently requires effective preventive strategies aimed at maximizing peak bone density, preventing excessive bone loss, and reducing the risk of falls. PMID- 18407105 TI - Disorders of steroid 11 beta-hydroxylase isozymes. AB - Steroid 11 beta-hydroxylase activity in the adrenal cortex is required for the synthesis of the major glucocorticoids and mineralocorticoids, but different isozymes mediate this conversion in the zona fasciculata, where cortisol is produced, and the zona glomerulosa, the site of aldosterone synthesis. The isozyme in the latter zone also has 18-hydroxylase and 18-oxidase activities that are required for aldosterone synthesis. Mutations in the genes encoding these isozymes respectively result in defective synthesis of cortisol and aldosterone. Recombinations between the two genes that alter the regulation of the isozyme responsible for aldosterone synthesis cause an inherited form of hypertension, glucocorticoid-suppressible hyperaldosteronism. PMID- 18407106 TI - Intraovarian regulation: peptidergic signaling systems. AB - Intraovarian regulation, an evolving field, is now at a crossroad. Although a number of putative intraovarian regulators appear to be of import to ovarian physiology, none has thus far been demonstrated to be indispensable to in vivo ovarian function. That notwithstanding, it is already clear that optimal gonadotropin hormonal action is highly contingent upon the input of tissue-based regulatory principles. It is with a strong sense of excitement that future work in this evolving area is anticipated. PMID- 18407107 TI - Management of cryptorchidism. AB - The forces guiding testicular descent have not been completely elucidated. Both testosterone and anti-Mullerian hormone might play a role. Available evidence suggests that malfunction of the testes of some sort usually precedes maldescent. The proper management of cryptorchidism has long been a controversial issue. In unilateral cryptorchidism, hormonal function and fertility are generally normal. To maximize fertility in patients with bilateral cryptorchidism, surgical treatment should be completed ideally by the first birthday. GnRH is unlikely to be of much help in initiating testicular descent. Cryptorchidism is associated with a three- to tenfold increase in testicular cancer. Twenty percent of tumors in unilateral cryptorchidism are in the normally descended testes. The condition of all boys and men with a history of cryptorchidism should be followed by physicians their entire lives, and these boys and men must become proficient in self-examination. PMID- 18407108 TI - Pituitary lactotrophs: endocrine, paracrine, juxtacrine, and autocrine interactions. AB - The synthesis and release of PRL by the lactotrophs is subjected to multiple regulators that are classified into four categories: endocrine, paracrine, juxtacrine, and autocrine. Endocrine agents originate from the hypothalamus, gonads, and the posterior pituitary. Paracrine factors are produced by cells of the intermediate and anterior lobes. Juxtacrine transmitters arise from extracellular matrix and cells adjacent to the lactotrophs. Autocrine agents are synthesized by the lactotrophs themselves. Consequently, the overall secretory activity of the lactotrophs reflects a balance between local and distant releasing and inhibiting factors. PMID- 18407109 TI - Evolving role of Gonadotropin-Releasing Hormone antagonists. AB - GnRH antagonists, unlike GnRH agonists, do not act via "downregulation." Instead, GnRH antagonists monopolize the GnRH receptors to such an extent that endogenous GnRH is unable to bind to sufficient numbers of GnRH receptors to provoke release of LH/FSH. This fundamental difference in the mechanism of action of GnRH antagonists versus GnRH agonists is anticipated to result in clinical benefits for certain applications. PMID- 18407110 TI - Urinary pyridinium crosslinks of collagen: specific markers of bone resorption in metabolic bone disease. AB - The hydroxypyridinium compounds pyridinoline and deoxypyridinoline are specific constituents of mature skeletal collagens. They are released into the circulation and excreted in the urine. Their measurement in urine is a sensitive index of the extent of ongoing bone resorption. Currently, quantification of collagen crosslinks in urine is achieved by chromatographic techniques, but more convenient immunoassays will make these measurements more widely available in the near future. Clinical applications of hydroxypyridinium markers include numerous metabolic bone disorders such as osteoporosis, primary hyperparathyroidism, Paget's disease of bone, and metastatic bone disease. Urinary pyridinium crosslinks of collagen also show great promise as markers of therapeutic efficacy in bone disorders associated with accelerated bone resorption. PMID- 18407111 TI - Prorenin in diabetes mellitus. AB - A rise in plasma prorenin often precedes the onset of vascular injury in patients with diabetes mellitus. Plasma prorenin measurements may be useful for predicting which patients will develop vascular injury and for monitoring the progression of the disease. A hypothesis is presented that accounts for these relationships and for the cosecretion of prorenin and renin into the circulation. PMID- 18407112 TI - Magnetic resonance imaging in patients with hypopituitarism. AB - In patients with hypopituitarism, magnetic resonance (MR) imaging of the hypothalamus and pituitary has disclosed a high incidence of hypoplasia of the anterior pituitary lobe, attenuation or transection of the pituitary stalk, and formation of an "ectopic" posterior pituitary lobe at the base of the hypothalamus. These anatomic abnormalities may be associated with other congenital malformations of the central nervous system, or may be due to an in utero toxic or infectious insult, perinatal trauma, neonatal asphyxia and hypoxia, head injury, or hemorrhage into a pituitary adenoma. The progressive development of defects in pituitary hormone secretion in such patients is probably due to continued atrophy of an anterior pituitary remnant with a limited vascular supply unstimulated by hypothalamic neuropeptides. By contrast, in patients with isolated hypogonadotropic hypogonadism, hypothalamic pituitary anatomy is normal, although abnormalities of the olfactory sulcus are present in patients with anosmia and hypogonadotropism (Kallmann syndrome). In most patients with central diabetes insipidus, the neurohypophysis is absent on MR scan. PMID- 18407113 TI - Receptors for pituitary adenylate cyclase-activating polypeptide: comparison with vasoactive intestinal peptide receptors. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a new member of the secretin glucagon-vasoactive intestinal peptide (VIP) family of peptides, being most homologous to VIP. PACAP exists in two amidated forms with 38 residues (PACAP38) and 27 residues (PACAP27), respectively. PACAP38 is the major form in tissues. There are two types of high-affinity receptors for PACAP: type I, which specifically binds to both PACAPs, and type II, which is shared with VIP. Type I PACAP receptors appear to have two subtypes: type IA, which binds to both PACAP38 and PACAP27, with slight preference for the latter, and type IB, with greater preference for PACAP38. Distribution of the type I PACAP receptor is different from that of VIP, and it is found in high concentrations in brain, spinal cord, anterior pituitary, adrenal medulla, spermatogonia at certain stages, mature spermatozoa, and some cell lines. Type II PACAP receptors are found in lung, liver, intestine, and other tissues, and their distribution is similar to that of the VIP receptor. Type II PACAP receptor might be similar to or identical with the VIP receptor. PMID- 18407114 TI - The role of brain angiotensin II in the regulation of Luteinizing Hormone and Prolactin secretion. AB - The renin-angiotensin system, both in the circulation and in the brain, is known for its role in the regulation of fluid balance and blood pressure. The brain angiotensin II (Ang II) system is also involved in the control of anterior pituitary hormone secretion, through affecting the secretion of releasing and inhibitory factors into the hypophyseal portal vessels. Ang II controls the release of LH and PRL in a manner that is modified by ovarian hormones, observed only under specific conditions, and localized to particular regions of the brain. The identification of Ang II systems in the pituitary gland and ovary, along with data showing effects of ovarian hormones on the activity of the brain Ang II system, suggests a feedback loop whereby the brain, pituitary, and gonads interact to affect reproductive function. PMID- 18407115 TI - Phosphate homeostasis, 1,25-dihydroxyvitamm D(3), and hyperparathyroidism in early chronic renal failure. AB - In early chronic renal failure, low plasma levels of calcitriol (1,25[OH](2)D(3)) do not seem to be merely the consequence of a reduced mass of functional nephrons. Indeed, this alteration can be considered as a compensatory mechanism, as analyzed according to a new concept of inorganic phosphate (P(i)) homeostasis that integrates both 1,25(OH)(2)D(3) production and renal P(i) reabsorption as essential regulating elements. Accordingly, the observed reduction in the renal production of 1,25(OH)(2)D(3) that occurs concomitantly with a decrease in tubular P(i) reabsorptive capacity (TmP(i)/GFR) may well represent a secondary adaptive response to a primary alteration in P(i) homeostasis. This crucial alteration in P(i) homeostasis would consist of an overload of a putative regulated intracellular P(i) pool, the localization of which remains to be determined. The observed hypophosphatemia, hypocalcemia, and PTH hypersecretion would represent alterations secondary to a low TmP(i)/GFR and to reduced 1,25(OH)(2)D(3) production. According to this pathophysiologic sequence, 1,25(OH)(2)D(3), rather than PTH as proposed in a former theory, would be "traded off" to preserve P(i) homeostasis in early chronic renal failure. Both theories predict that dietary P(i) restriction represents a logical preventive therapy at least until the nature of the primary defect in P(i) homeostasis is understood. However, assuming that low 1,25(OH)(2)D(3) levels in early chronic renal failure represent a compensatory phenomenon, this new theory suggests that calcitriol should be only administered at a later stage of the disease, when the production of 1,25(OH)(2)D(3) becomes inappropriately low to maintain mineral homeostasis. PMID- 18407116 TI - The role of glucocorticoids in the regulation of Growth Hormone secretion: mechanisms and clinical significance. AB - Glucocorticoids are well known to inhibit growth and GH secretion in humans and animals, yet in vitro these steroids stimulate GH synthesis and secretion. These opposite actions appear to be mediated at different sites. The inhibition involves modulation of hypothalamic somatostatin and the stimulation involves direct actions on the pituitary. Current evidence suggests that the predominant action in vivo is through the inhibitory influences of somatostatin. PMID- 18407117 TI - Hypothalamic-pituitary sarcoidosis. AB - Patients with sarcoidosis may develop hypopituitarism secondary to granulomatous infiltration of the pituitary and hypothalamus. All degrees of anterior pituitary insufficiency can occur, ranging from selective deficiency to panhypopituitarism; diabetes insipidus occurs frequently. Commonly associated neurologic manifestations are cranial neuropathies, aseptic meningitis, and visual field defects. Although neurologic deficits respond well to corticosteroids, hormonal abnormalities generally persist despite therapy. PMID- 18407118 TI - Steroid receptor folding by heat-shock proteins and composition of the receptor heterocomplex. AB - Over the past 2 years, reports from several laboratories have supported the proposal that the steroid receptors are bound through the hormone-binding domain to a protein complex that contains three heat-shock proteins-hsp90, hsp70, and hsp56. This receptor-heat-shock-protein heterocomplex accounts for the behavior of the classic 9 S, non-DNA-binding form of the adrenocorticoid, sex hormone, and dioxin receptors. The receptor heterocomplex has now been reconstituted by an enzymatic system in reticulocyte lysate. This represents the first in vitro system for reversing receptor transformation, and this ability to reconstitute the receptor heterocomplex promises rapid advances in our understanding of how these receptors are folded, transported, and regulated by hormone in the cell. PMID- 18407119 TI - Bovine placental prolactin-related hormones. AB - The bovine placenta, like that of rodents and primates, synthesizes members of the PRL/GH gene family, which may assist the pituitary hormones or perform unique functions during pregnancy. Bovine placental lactogen (bPL) potentially may act through.three receptors: as an agonist or partial antagonist at the PRL and GH receptors, and via an apparently specific receptor in the endometrium. A large distinct subfamily of diverse primary structure, including bovine PRL-related protein I (bPRP-I), evidently does not act via these receptors. Advances in our understanding of hormone-receptor interactions for this gene family have provided new tools to study the role of these hormones in the successful pregnancy. PMID- 18407120 TI - Evolution of gonadotropin-releasing hormones. AB - GnRH was originally isolated as a hypothalamic peptide hormone that regulates the reproductive system by stimulating the release of gonadotropins from the anterior pituitary. However, multiple molecular forms of the peptide have evolved, which have been coopted for a variety of regulatory functions: as a neurotransmitter in the central and sympathetic nervous systems, as a paracrine regulator in the gonads and placenta, and as an autocrine regulator in tumor cells. We review here the evolution of these variant forms of GnRH and their functions. PMID- 18407121 TI - G protein oncogenes in pituitary tumors. AB - G proteins are involved in the transduction of external signals from cell surface receptors to intracellular effectors. Somatic mutations activating the a-subunit of G(s) (the stimulatory regulatory protein of adenylyl cyclase) by inhibiting its intrinsic GTPase activity have been first identified in human GH-secreting adenomas and subsequently found in thyroid tumors and in McCune-Albright syndrome. It has been therefore proposed that the gene encoding the GS a-subunit may be converted into an oncogene (gsp for GS protein) in cell types that proliferate in response to cAMP. Since several G proteins mediate signaling pathways that are effective in coupling external stimuli to cell proliferation, it appears most likely that in the near future other G protein oncogenes will be identified in human tumors. PMID- 18407122 TI - Functional cross-talk between receptors for peptide and steroid hormones. AB - Communication between a cell surface peptide hormone receptor and an intracellular steroid hormone receptor can take various routes, as dictated by the physiology of a particular cell type. There is increasing evidence for a novel route which requires that a peptide hormone receptor pathway converge on a steroid hormone receptor, leading to its activation. One consequence of such a process can be signal amplification for the peptide hormone receptor agonist. This is exemplified by the self-potentiating action of GnRH, which is a critical component in events leading to a surge in LH secretion and ovulation. One signaling pathway stimulated by the GnRH receptor may entail a phosphorylation cascade resulting in progesterone-independent modulation of progesterone receptor activity. PMID- 18407123 TI - Hypoglycemia in infants. AB - Because the infant's brain is to a large extent dependent on glucose utilization, hypoglycemia of infants can have grave effects on brain function, and it is important to diagnose it and, when possible, treat it promptly. Causes of hypoglycemia in infants are (a) excess insulin secretion, (b) factitious hyperinsulinemia, (c) GH or ACTH deficiency, (d) primary glucocorticoid deficiency, (e) defects of the enzymes involved in hepatic glucose production, or (f) defects in hepatic fatty acid oxidation. PMID- 18407124 TI - Aluminum and renal osteodystrophy A diminishing clinical problem. AB - Aluminum-associated osteomalacia is a serious complication of advanced renal disease. Sources o f aluminum include the water used for hemodialysis and the gastrointestinal absorption o f aluminum from the antacids used to bind dietary phosphorus in an effort to prevent hyperparathyroidism. Definitive diagnosis is made by bone biopsy and staining for aluminum. Noninvasive testing using the deferoxamine challenge test together with serum PTH levels is also of value in many instances. The goal for the future is to eliminate aluminum-related osteodystrophy by the avoidance of aluminum, control of phosphorus with non aluminum-containing binders, and development of novel approaches to facilitate phosphorus removal by dialysis. PMID- 18407125 TI - Management of asymptomatic hyperparathyroidism A report on the NIH consensus development conference. PMID- 18407126 TI - Progesterone receptor phosphorylation complexities in defining a functional role. AB - All steroid receptors are phosphoproteins and several, including progesterone receptors (PRs), become hyperphosphorylated upon binding of ligand. PR phosphorylation is complex, occurring in different cellular compartments and perhaps requiring multiple serine kinases. A model that is emerging proposes that PR phosphorylation is progressive, occurring in at least a three-stage cascade. However, the functional significance of this phosphorylation cascade remains unclear. PMID- 18407127 TI - Molecular basis of the X-chromosome-linked Kallmann's syndrome. AB - Kallmann's syndrome combines hypogonadotropic hypogonadism and anosmia. The most frequent form of the disease is linked to the X chromosome and has been proposed to be due to a defect in the embryonic migration of GnRH neurons and olfactory axons from the nose to the brain. A candidate gene for the X-linked form of the disease has been isolated by positional cloning. Mutations in the open reading frame have been identified in several patients, providing convincing evidence that this gene is the actual gene, KAL, responsible for the X-linked Kallmann's syndrome. Correlations between molecular and clinical data extend the role of the KAL gene to other neuronal pathways and kidney organogenesis. The deduced amino acid sequence led us to postulate that the KAL protein is an extracellular matrix component with possible antiprotease and adhesion functions. Such functions are known to be involved in neuronal migration, axonal guidance and targeting, and also in synaptogenesis. Further experiments will enable the elucidation of the role of the KAL protein. PMID- 18407128 TI - Granins markers of the regulated secretory pathway. AB - The granins are a family of acidic secretory proteins made up of chromogranin A, chromogranin B, and secretogranin II, which exhibit widespread distribution in endocrine and neuronal cells. The numerous potential sites for proteolytic processing have suggested a role for these peptides as prohormones: several potential degradation products of chromogranin A, pancreastatin, and chromostatin have autocrine activity. On the other hand, an intracellular role for the granins is supported by their propensity to aggregate in a low-pH, high-calcium environment such as found in the trans-Golgi network followed by their efficient sorting to the regulated pathway. As a result, the granins are considered markers for the regulated pathway and may play a role in secretory granule formation. PMID- 18407129 TI - Role of bisphosphonates in metabolic bone diseases. AB - Bisphosphonates are potent inhibitors of osteoclast-mediated bone resorption and represent the treatment of choice in disorders associated with accelerated bone resorption. They are widely used in the management of Paget's disease of bone and hypercalcemia of malignancy. Over the past few years, evidence has been accumulating for a beneficial effect of these agents in tumor-induced osteolytic bone disease and in established osteoporosis. Bisphosphonates may also play a useful though limited role in the medical management of hyperparathyroidism and heterotopic ossification. PMID- 18407130 TI - Hormonal control of thermogenesis and energy dissipation. AB - Facultative (adaptive) thermogenesis is primarily controlled by the sympathetic nervous system (SNS). The participation of thyroid hormones in adaptive thermogenesis has been considered minor or, at most, permissive. The finding of type II-thyroxine (T(4)) 5'-deiodinase in brown adipose tissue (BAT) has opened a way to uncover a more important role for thyroid hormone in adaptive thermogenesis. This enzyme is activated by the. SNS and insulin. When activated, it generates high BAT concentrations of triiodothyronine (T(3)) from plasma T(4). T(3), intrinsically 10 times more active than T(4), has been shown essential for the expression of the key protein in BAT thermogenesis, uncoupling protein (UCP). The multihormonal control of BAT type-II 5'-deiodinase and the marked influence of T(3) on UCP and BAT thermogenesis suggest that the local control of T(3) generation may be an important source of variability in the potential of mammals to maintain temperature and dissipate energy. PMID- 18407132 TI - Clusterin Insights into a multifunctional protein. PMID- 18407131 TI - Fifth international conference of the pituitary pathologists' club, 17-22 September 1992, Toronto, and Hockley Valley Resort, Orangeville, Ontario, Canada. PMID- 18407133 TI - Transcriptional regulation of the adrenal steroidogenic enzymes. AB - Corticosteroid biosynthesis requires the concerted action of a related group of cytochrome P450 steroid hydroxylases. The genes encoding these steroid hydroxylases exhibit two distinct levels of transcriptional regulation: selective expression in steroidogenic cells and induction in response to trophic hormones. With respect to cell-selective expression, recent studies have identified a nuclear receptor protein expressed only in steroidogenic cells that is postulated to regulate the expression of all cytochrome P450 steroid hydroxylases through common promoter elements. In contrast, the coordinate responses of these genes to trophic hormones are not readily explained by a unifying mechanism, and their hormone responsive expression probably involves multiple promoter elements. PMID- 18407134 TI - A new look at an old hormone gastrin. AB - The origin of gastrin from the G cell has been identified, as have a number of mechanisms regulating both secretion and processing of the peptide. Little, however, is known of the intracellular regulation of processing and secretion. Gastrin receptors have been identified and cloned, although the exact relationship to the cholecystokinin receptor remains to be clarified. The gene for the peptide has been sequenced and various promoters identified. Up- and downstream sequences have been demonstrated to be interrelated with epidermal growth factor and somatostatin, and the existence of a complex gastric-based molecular regulatory system is apparent. The dynamic role of gastrin in the regulation of acid secretion has been delineated by its action on multiple targets in gastric parietal cells and endocrine enterochromaffinlike cells. Similarly, its trophic effects on gastrointestinal tissue have been explored, resulting in its identification as a common denominator of gastric fundic hyperplasia and microcarcinoid evolution in conditions of hypergastrinemia. PMID- 18407135 TI - The molecular basis of a hereditary form of hypertension, glucocorticoid remediable aldosteronism. AB - Glucocorticoid-remediable aldosteronism (GRA) is a hereditary cause of human hypertension in which aldosterone secretion is regulated by ACTH. Recent studies demonstrate that this disorder is caused by fusion of regulatory sequences of the steroid 11beta-hydroxylase gene to coding sequences of the aldosterone synthase gene. These mutations occur by unequal crossing over between these two genes and result in ectopic expression of aldosterone synthase in adrenal fasciculata. These features explain the physiology and genetics of GRA and provide the basis for a simple direct genetic test for this disorder. PMID- 18407136 TI - Stress update Adaptation of the hypothalamic-pituitary-adrenal axis to chronic stress. AB - The hypothalamic-pituitary-adrenal (HPA) axis exhibits a circadian rhythm, activation by stress, and inhibition by corticosteroids. Activity in the HPA axis is very sensitive to inhibition by corticosteroids when they are administered exogenously. When stress-induced corticosteroid secretion occurs, however, normal activity in the HPA is not inhibited and may even be augmented. Experiments in rats have shown that stress also induces facilitation of subsequent activity in the HPA axis that appears to balance the inhibitory effects of corticosterone and thus maintains responsiveness to new, acute stresses in chronically stressed rats. Stress-induced facilitation of HPA axis activity may be mediated by a parallel stress-induced (CRH-dependent) increase in the capacity of brain noradrenergic cell groups to respond to acute stress. A continually responsive HPA axis, even under conditions of chronic stress, appears to be important for survival. Stress-induced increases in glucocorticoid secretion to levels sufficient to occupy glucocorticoid receptors enable appropriate thermoregulatory and cardiovascular responses to acute stress. There is, however, an overall metabolic cost to the animal of maintaining continued activity in the HPA axis during chronic stress. PMID- 18407137 TI - Steroid sulfation Current concepts. AB - Formation of steroid sulfates is catalyzed by sulfotransferase enzymes that are widely distributed and frequently of high specificity. Steroid sulfates cannot be described as being active hormones, but some serve in tissue sites as precursors of active steroids formed by enzymic cleavage of the sulfate group by sulfatase enzymes. There is increasing evidence that intracellular sulfation and desulfation can play a role in regulating the availability of active steroid hormones near target sites. There are strong indications for this regulation in the uterine compartment, in the liver, and in mammary cancer cells. PMID- 18407138 TI - Pit-1 and hypopituitarism. AB - The story of Pit-1 and hypopituitarism in humans provides an excellent example of pleiotrophism or multiple phenotypic effects resulting from a single genetic alteration. It shows how defects in this single gene cause the absence o f several pituitary hormones. Three recent articles reviewed here provide examples of different mutations in this homeobox gene encoding a transcriptional activation protein that is vital to the embryologic development, survival, and differentiated function of somatotropes, lactotropes, and thyrotropes. PMID- 18407139 TI - Mutations of the human glucokinase gene and diabetes mellitus. AB - The enzyme glucokinase catalyzes the phosphorylation of glucose and plays a key role in the regulation o f insulin secretion by pancreatic beta cells and glucose disposal in hepatocytes. Recent studies have shown that mutations in the gene encoding this key regulatory enzyme of glycolysis are a common cause of an autosomal dominant form of non-insulin-dependent (type 2) diabetes mellitus that has an onset often during childhood. The association of mutations in the glucokinase gene with impaired pancreatic cell function underscores the importance of glycolysis in the regulation of insulin secretion and suggests that mutations in other genes expressed in the beta-cell that also control rate limiting steps in glucose metabolism may lead to diabetes. PMID- 18407140 TI - Circulating binding proteins for the insulinlike growth factors. AB - Six distinct insulinlike growth factor-binding proteins (IGFBP-1 to IGFBP-6) of core molecular mass 20-30 II have been identified, of which one, IGFBP-3, carries most o f the circulating IGF-I and IGF-II in ternary complexes that also contain an acid-labile glycoprotein subunit. Although the role of circulating IGFBP-3 in IGF stabilization and transport is becoming increasingly well understood, the functions of the other IGFBPs in the circulation are less clear, and some redundancy of function is possible. IGFBP-l, which is metabolically regulated by insulin and carbohydrates, may act as a counterregulator in blood glucose regulation and could also be important in targeting IGF delivery from the circulation to the tissues, while recent studies of IGFBP-2 physiology suggest that this protein may function as an additional IGF carrier when IGFBP-3 levels are inadequate. Research into the roles of circulating IGFBP-4, -5, and -6 is, as yet, poorly developed and will depend on the ready availability of analytical methods for these proteins. PMID- 18407141 TI - Regulation of insulin gene transcription. AB - Selective transcription of the insulin gene in pancreatic beta cells is regulated by its enhancer, located between nucleotides -340 to -91 relative to the transcription start site. The activity o f the enhancer is controlled by both positive- and negative-acting cellular factors. Cell-type-specific expression is mediated principally by a single cis-acting enhancer element, termed the insulin control element (ICE), which is acted upon by both these cellular activities. This review focuses on the role of the factors acting on the ICE and other enhancer control elements in the establishment of cell-type-specific and physiologically regulated transcription of the insulin gene. PMID- 18407142 TI - Immunotherapy at clinical diagnosis of insulin-dependent diabetes: an approach still worth considering. AB - Immunointervention at the time of clinical diagnosis of insulin-dependent diabetes has been adopted in many centers throughout the world, with the aim of inducing and maintaining clinical remission (for example, reduction of insulin replacement therapy with normalization of metabolic parameters). Several approaches have been considered, ranging from the use of different kinds of immunosuppression to the use of free oxygen radical scavengers. Although the period of latency before the onset of overt hyperglycemia should be regarded as the most favorable time for any sort of immunointervention, adjuvant treatment at diagnosis (in addition to intensive insulin therapy) may offer clinical advantages in the long term. Thus, maintenance of some residual beta-cell function as a result o f such an approach may improve metabolic control and help to prevent the insurgence of late diabetic complications. PMID- 18407143 TI - Growth hormone-insulinlike growth factor I and immune function. AB - Growth hormone (GH) and insulinlike growth factor I (IGF-I) may be part of a neuroendocrine immune axis that stimulates cellular proliferation of primary lymphoid organs (bone marrow, thymus) as well as stimulates activation of peripheral lymphocytes and macrophages to enhance specific immune responses. GH can also stimulate production of thymic hormones and cytokines, and in this way impact on immune function. It is not clear whether GH and IGF-I act independently or whether the action of GH is mediated by local production of IGF-I by lymphocytes. Both GH and IGF-I and their receptors are present in lymphocytes. Thus, cells of the immune system may be important targets of the GH-IGF-I axis. PMID- 18407144 TI - Investigation of Cushing's syndrome by inferior petrosal sinus sampling. AB - Bilateral simultaneous inferior petrosal sinus sampling, in combination with CRH stimulation, is now able to confirm the diagnosis of pituitary-dependent Cushing's disease with near certainty. In expert hands, the procedure is straightforward and safe. As well as confirming the differential diagnosis, the test may aid surgical success, especially if no adenoma is apparent at transsphenoidal exploration. In this article, we review the technique and its interpretation and consider which patients should undergo the procedure. PMID- 18407145 TI - Pheochromocytoma: 1926-1993. AB - Pheochromocytoma is a tumor frequently sought and rarely found. It is associated with spectacular cardiovascular disturbances and, when correctly diagnosed and properly treated, it is curable; when undiagnosed or improperly treated, it can be fatal. Catecholamine-producing tumors that arise from chromaffin cells of the adrenal medulla and sympathetic ganglia are termed pheochromocytomas and paragangliomas, respectively. However, the term pheochromocytoma has become the generic name for all catecholamine-producing tumors. The biochemical diagnosis is straightforward. The localization of pheochromocytoma has been greatly facilitated by advances in computerized imaging and meta-iodobenzylguanidine scanning. Treatment with preoperative alpha- and beta-adrenergic blockade followed by surgical excision of the pheochromocytoma are associated with very low morbidity and mortality. PMID- 18407146 TI - Amiodarone and thyroid dysfunction. AB - Amiodarone is a potent and widely used antiarrhythmic drug that bears a structural resemblance to thyroid hormones. The high iodine content of the drug determines that amiodarone induces changes in circulating concentrations of thyroid hormones, largely through inhibition of conversion of thyroxine (T(4)) to tri-iodothyronine (T(3)). Amiodarone treatment typically results in a rise in serum T(4), often to above the normal range, associated with a fall in circulating T(3). These biochemical changes are found in subjects who remain clinically euthyroid. In addition to changes in circulating thyroid hormones found in euthyroid subjects, overt thyrotoxicosis and hypothyroidism may complicate amiodarone treatment. Amiodarone-induced thyrotoxicosis is more common in areas of iodine deficiency, whereas hypothyroidism is more common in iodine rich parts of the world. PMID- 18407147 TI - Gastrointestinal peptides and cancer. AB - The term gastrointestinal peptide is usually used to refer to a group of peptides with relatively low molecular weight (generally <30 amino acids) that function as agonists by interacting with cell surface receptors on one or more gastrointestinal target cells. These peptides can function as hormones or neurotransmitters, or can act as paracrine regulators of target cell function. Recently it has become apparent that some gastrointestinal peptides may play important roles in influencing the rates of proliferation of certain neoplastic cells. In the present article, we highlight major areas of research that provide information about the relationships between gastrointestinal peptides and cancer. In addition to identifying what we believe is a "trend," we also make suggestions as to likely future developments. PMID- 18407148 TI - Models of human endocrine disorders in transgenic rodents. AB - Although the primary etiology of some endocrine diseases is obvious, for many of them the initiating abnormality is not known. Usually this uncertainty is due to a slow development of significant clinical symptoms, combined with the fact that an abnormality in any one endocrine gland will lead to secondary endocrine abnormalities. The ability to introduce relatively specific and (sometimes) subtle genetic changes into rodents provides an opportunity to test hypotheses concerning primary events in human endocrine disorders. This review deals with the development of transgenic rodent models of the more common, classically defined human endocrine abnormalities. PMID- 18407149 TI - Prolactin and autoimmune disease. AB - A number of reports have shown that PRL is an immune-stimulating hormone that is capable of stimulating organ-specific inflammatory disease in humans. More recently, hyperprolactinemia has been associated with the active phase of the immune-complex-mediated autoimmune disease, systemic lupus erythematosus. The theory that PRL contributes substantially to disease activity was upheld in the NZB/W mouse model of spontaneous, hormone-sensitive lupus. Implanted pituitary glands resulted in hyperprolactinemia, accelerated proteinuria, high levels of circulating IgG, and premature death. Therapeutic studies with NZB/W mice, as well as anecdotal evidence from a small number of patients, have provided evidence that PRL suppressive therapy may be beneficial in selected cases of autoimmune disease. PMID- 18407150 TI - Thyroid hormone regulation of Na,K-ATPase expression. AB - Active Na,K transport across plasma membranes (mediated by Na,K-ATPase) is stimulated by triiodothyronine (T(3)) in all mammalian tissues responsive to thyroid hormone, and this stimulation has been proposed to account for a substantial fraction of thyroid thermogenesis. The enhancement of Na,K-ATPase activity by T(3) results from increased biosynthesis of Na,K-ATPase subunits and is associated with increased abundance of their encoding mRNAs. In certain target tissues, T(3) preferentially augments the expression of the alpha2 isoform of the enzyme (characterized by its high sensitivity to inhibition by cardiac glycosides). The T(3)-induced increase in Na,K-ATPase subunit mRNA expression has been shown to be mediated by both transcriptional and post-transcriptional mechanisms. PMID- 18407151 TI - Regulation of retinoid and thyroid hormone action through homodimeric and heterodimeric receptors. AB - Biologic responses to retinoids and thyroid hormones are mediated by their intracellular receptor proteins. Many exciting advances have been made recently in understanding the molecular mechanism by which these receptor proteins operate. In contrast to the steroid hormone receptors that function predominantly as homodimers, thyroid hormone receptors (TRs)and retinoic acid receptors (RARs) require interaction with the retinoid X receptors (RXRs) for efficient DNA binding and transactivation. In addition, RXRs, in the presence of their specific ligands such as 9-cis RA, can form homodimers that recognize a subset of retinoic acid responsive elements (RAREs). The retinoid responses mediated by RXR homodimers and RAR-RXR heterodimers can be restricted by the COUP-TF orphan receptors that bind strongly to certain RAREs as homodimers. Thus, a complex network of receptor interaction has been unraveled that promises a better understanding of thyroid and retinoid hormone regulation of fundamental biologic processes and diseases. PMID- 18407152 TI - Opioids in neural and nonneural tissues. AB - The endogenous opioid peptides (EOP) are grouped in three families, each deriving from the posttranslational processing of a distinct precursor molecule and exhibiting high affinity for a specific opioid receptor. The genes of EOPs are expressed in a wide variety of sites, including many nerve, neurosecretory, and endocrine cells. In reviewing the vast literature on this subject, a few patterns begin to emerge. First, the distribution of EOPs in tissues appears to be a distinct characteristic of each family of opioids. Second, the EOP producing cells can be grouped into two broad categories: those expressing only one and those expressing multiple EOP genes. Most EOP-producing nerve and neurosecretory cells fall into the first category, that is, they express one EOP gene, whereas most nonneural cells fall into the second category, that is, they express multiple EOP genes. Third, it appears that there is a relationship between opioids, proliferation rate, and state of differentiation of cells, since it has been shown that (a) mitogenic factors may change the EOP profile of a cell, and that (b) opioids may inhibit the proliferation rate of normal or neoplastic cells. The physiologic implication of these observations is briefly discussed. PMID- 18407153 TI - Influence of thyroid hormones on the GH-IGF-I axis. AB - There is a complex relationship between thyroid hormones, GH, and the insulin like growth factors (IGFs). Thyroid hormones act at many sites from the hypothalamic control of GH release to the tissue expression of IGF-I and its binding proteins (IGFBPs). In this review, we present current knowledge of the effects of altered thyroid status on the GH-IGF-I axis, concentrating on the changes seen in IGF-I gene expression and circulating levels of GH and IGFBPs. PMID- 18407154 TI - Parathyroid hormone-related protein a peptide of diverse physiologic functions. AB - Parathyroid hormone-related protein (PTHrP) is the factor responsible for the syndrome of humoral hypercalcemia of malignancy. PTHrP is produced by a multitude o f normal as well as malignant cells, and exerts both classic parathyroid hormone (PTH)-like and PTH-unlike effects. The molecular cloning of the PTHrP gene, and the subsequent recognition of its widespread expression in normal tissues under normal physiologic conditions, has prompted intense inquiry into its biologic function. PTHrP appears to act in an autocrine or paracrine fashion in (a) normal embryogenesis and neonatal development, (b) cellular growth and differentiation, (c) reproduction and lactation, (d) epithelial calcium transport, and (e) smooth muscle relaxation. These five key emerging physiologic roles of PTHrP are the focus of this review. PMID- 18407155 TI - Does inhibin have an endocrine function during the menstrual cycle? AB - Inhibin (alpha-beta heterodimer) has been considered to be the principal nonsteroidal ovarian regulator of pituitary FSH secretion. The beta-beta heterodimer, activin, produced by the ovary and other tissues, appears to act locally, with actions opposite to those of inhibin. Since immunoreactive inhibin is highest during the luteal phase of the menstrual cycle when FSH is lowest, a negative feedback role in controlling FSH release at this time has been suggested. Attempts to establish this by using immunoneutralization techniques have failed to reveal such a role. We must enhance our understanding of the gonadotropic control of inhibin/activin gene expression within the various compartments of the primate ovary, the role of their binding proteins, and the nature of the secretory products before we can resolve the question of whether inhibin has an endocrine function during the menstrual cycle and how cyclic reinitiation of follicular development is controlled. PMID- 18407156 TI - Lipoprotein-thyroid hormone interactions. AB - A small but significant portion of the thyroid hormones that circulate in human plasma is associated with lipoproteins. Although the major lipoprotein carrier is HDL, the role of these interactions on T(4) entry into cells was tested first with LDL and human fibroblasts because of the well-characterized LDL receptors and the availability of cells with genetically absent receptors. It was shown that LDL enhanced the uptake of T(4) by cultured fibroblasts in which the LDL receptors were expressed. The T(4) binding sites on apolipoproteins B-100 and E, as well as apoA-I, have been partially characterized, and they exhibit considerable homology. A number of possible physiologic consequences of thyroid hormonelipoprotein interactions have been put forward as topics for further investigation. PMID- 18407157 TI - The 3beta-hydroxysteroid dehydrogenase gene family of enzymes. AB - It now is apparent that a family of closely related genes encode for 3beta- hydroxysteroid dehydrogenase (3betaHSD). Studies on the regulation of these genes are in their infancy, but the regulation appears multifactorial. The various 3betaHSD genes are expressed principally in a tissue-specific manner likely involving separate mechanisms of regulation. To date, two human 3betaHSD genes and their products have been characterized; type I is expressed in placenta, sebaceous glands, and several other nonendocrine tissues, whereas the type II isoform is the principal 3betaHSD of adrenal cortex and gonads. PMID- 18407158 TI - Role of cyclic AMP in the control of cell-specific gene expression. AB - Genes have to be expressed in specific cell types at appropriate times of development dependent on external signals. cAMP signaling occurs in all cells, thus raising the question of how this signal transduction pattern is integrated into mechanisms determining cell-specific gene expression. We have analyzed expression of the tyrosine aminotransferase gene as a model to study the basis of this cell type specificity of hormone induction. We found that cell-type-specific expression is generated by combined action of cAMP signal-dependent and liver cell-specific transcription factors. The interdependence of the cAMP response element and an element determining liver cell specificity enables a gene to respond to an ubiquitous signal in a cell-specific manner. PMID- 18407159 TI - Neuropeptidergic regulation of feeding behavior Neuropeptide Y. AB - Neuroendocrine control of feeding behavior is multifactorial, involving a variety of peripheral and central signals. Neuropeptides, catecholamines, and serotonin constitute the signals of the feeding circuitry acting primarily in the brain, especially at the hypothalamic level. In this review, an attempt is made to summarize the recent progress made in our continuing effort to understand the regulation of feeding behavior by neuropeptides, particularly those that stimulate feeding. A special emphasis has been placed on a neuropeptide of the pancreatic polypeptide family-neuropeptide Y (NPY). PMID- 18407160 TI - Molecular genetics of thyroid cancer. AB - Tumors of the human thyroid follicular cell demonstrate multiple "routes" and multiple stages of development, offering an unparalleled opportunity for correlating clinicopathologic tumor behavior with the underlying molecular genetic abnormalities. This review summarizes the clinical and experimental evidence supporting the causal role of five key genes in thyroid oncogenesis, namely, the oncogenes ras, gsp, ret, and trk, and the tumor-suppressor gene TP53. The nature of the somatic mutations is described and the likely mechanisms discussed by which they perturb cellular growth signal transduction to produce particular pathologic phenotypes. A model of thyroid oncogenesis is presented that suggests that the pattern of tumor development is determined by the nature of the initiating oncogenic event. PMID- 18407161 TI - Detection and management of early diabetic nephropathy. AB - It has proven difficult to alter the progression of diabetic nephropathy once overt proteinuria is established. The presence of microalbuminuria reflects an early renal lesion that may be more amenable to therapeutic intervention. Dietary protein restriction, improved glycemic control, and aggressive treatment of high blood pressure all have shown beneficial effects in some patients. Angiotensin converting enzyme inhibitor therapy may offer specific advantages in terms of its renal protective effects. PMID- 18407162 TI - McCune-Albright syndrome. AB - McCune-Albright syndrome (MAS) is characterized by the clinical triad of polyostotic fibrous dysplasia, cafe-au-lait pigmented skin lesions, and multiple endocrinopathies. The molecular basis of MAS is a mutation in G(s)alpha that results in constitutive activation of adenylyl cyclase in affected tissues. This mutation occurs during early embryogenesis, and therefore patients with MAS are mosaic. The identification of activating mutations of Gsa in liver, heart, and gastrointestinal tract of patients with MAS suggests a broader spectrum of clinical disease than previously appreciated. PMID- 18407163 TI - Molecular genetics of the ACTH and melanocyte-stimulating hormone receptors. AB - The related ACTH and melanocyte-stimulating hormone (MSH) receptors control adrenal steroidogenesis and pigmentation in response to an overlapping set of peptides derived from the proopiomelanocortin (POMC) molecule. The recent cloning of these receptors has already opened up a new understanding of their role in normal and pathologic functioning of the adrenal cortex, and of the process of pigmentation. The murine MSH receptor maps to a genetic locus called extension, a locus known since early in this century to control the relative amounts of the two major types of melanins: eumelanin and phaeomelanin. The highly variable pigmentation phenotypes resulting from different extension locus alleles are caused by structural mutations in the MSH receptor that alter the degree of its signal-transducing capacity. A mutation in the ACTH receptor in a patient with ACTH resistance has also recently been reported. It is likely that the etiology of this rare disease includes mutations that affect the functioning of the ACTH receptor. PMID- 18407164 TI - International symposium on glucagonlike peptide I, Copenhagen, Denmark, 17-19 May 1993. PMID- 18407166 TI - Corticostatins. AB - Corticostatic (anti-ACTH) peptides were first isolated from rabbit fetal and adult lung and from rabbit neutrophils. They have now been isolated from human, rat, and guinea pig tissues. They have several biologic activities that include inhibition of ACTH binding, stimulation of L-type Ca(2+) channels, monocyte chemotaxis, and degranulation of mast cells with the release of histamine. Rabbit corticostatin I (CSI) is widely distributed in maternal and fetal rabbit tissues and is localized to neutrophils, macrophages, and monocytes, and the changing concentration of CSI with advancing gestation indicates a possible physiologic role during pregnancy. PMID- 18407165 TI - Amylin in bone conservation current evidence and hypothetical Considerations. AB - Amylin, a 37-amino-acid long single-chain polypeptide, is structurally homologous to calcitonin and calcitonin gene-related peptide (CGRP). The peptide is secreted from pancreatic beta cells and is thought to have an anti-insulin action. Here, we review the recently described effects of amylin on calcium homeostasis and discuss its possible role in bone conservation. Amylin is a potent hypocalcemic and antiresorptive peptide. Studies using isolated osteoclasts have revealed that amylin inhibits cell motility (Q effect), without affecting cell spread area or elevating cytosolic [Ca(2+)]. Thus, amylin action is similar to that of calcitonin, but lower in potency. Lower circulating concentrations of amylin in type-1 diabetes may cause the bone loss associated with this condition. PMID- 18407167 TI - Perifused pituitary cells as a model for studies of gonadotropin biosynthesis and secretion. AB - Pituitary gonadotropes are influenced by a variety of hormones in vivo. In an effort to study these signals individually, several experimental models have been devised. One of these models, pituitary perifusion, has the dual advantage of removing endogenous hormones and growth factors and permitting exogenous signals to be applied in a continuous or pulsatile fashion. Application of the perifusion model to the study of gonadotropen biosynthesis and secretion has led to the discovery of several previously unappreciated interactions between hypothalamic and paracrine signals that act on gonadotropes. PMID- 18407168 TI - Pathogenesis of thyroid-associated ophthalmopathy. AB - Thyroid-associated ophthalmopathy is a complex disease whose pathogenesis is thought to be autoimmune. Evidence has accumulated to implicate lymphocytes, antibodies, and cytokines targeting the orbital tissues whose structural and functional characteristics are unique. Epitope sharing between the orbit and the thyroid is the likely explanation for the close association of ophthalmopathy with autoimmune thyroid disease. Environmental and other nonautoimmune factors are identified that also play a part in the initiation or perpetuation of the disease process. PMID- 18407169 TI - The DCCT-Finally! PMID- 18407170 TI - Current therapeutic approaches to hyperthyroidism. AB - Treatment of hyperthyroidism varies, depending upon the underlying etiology of thyrotoxicosis. Antithyroid drugs may be administered to patients with Graves' hyperthyroidism for prolonged periods in an attempt to obtain a remission. Alternatively, a short course of antithyroid drugs may be administered to patients with Graves' disease or toxic adenoma to achieve euthyroidism prior to definitive therapy with radioiodine or surgery. Radioiodine is the preferred therapy for most adult patients, however, large goiters, especially i f obstructive symptoms are present, are best treated with surgery. PMID- 18407171 TI - Human chorionic gonadotropin Molecular forms, detection, and clinical implications. AB - Different molecular forms of human chorionic gonadotropin (hCG) have been identified in biologic fluids of patients with various physiopathologic processes. These materials include (a) the intact heterodimer hCG comprising two mature a and beta subunits, and (b) the uncombined or free forms of the a (hCGalpha) and beta subunit (hCGbeta), and several fragments o f hCG such as the nicked forms o f both hCG and free hCGbeta and its ending degradation product, the beta-core fragment or hCGbetacf The determination of hCG and related molecules in biologic fluids is usually achieved by immunologic procedures, but discrepancies among kits remain a problem in clinical practice. Specific measurements of hCG and of, independently, its free beta subunit are important in the diagnosis and follow-up of either trophoblastic diseases or testicular cancers, whereas only the free hCGbeta has to be assayed for detection in nongonadal and nonplacental tumors. PMID- 18407172 TI - Peroxisome proliferator-activated receptors A link between endocrinology and nutrition? AB - Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily like the steroid, thyroid, or retinoid hormone receptors, which are ligand-activated transcription factors regulating gene expression. PPARs mediate the induction of the enzymes of the peroxisomal and microsomal fatty-acid oxidation pathways by hypolipidemic drugs such as clofibrate and are probably also involved in the gene expression of other lipid metabolism-associated proteins that are controlled by fibrate hypolipidemic drugs. That PPARs play an important role in the regulation of lipid metabolism is reinforced by the discovery of their activation by physiologic concentrations of fatty acids. This observation raises the question of whether fatty acids are ligands of PPARs, which would imply that nutritional fatty acids can act like hormones. PMID- 18407173 TI - A dialogue on analogues Newer vitamin-D drugs for use in bone disease, psoriasis, and cancer. AB - Many new analogues of the vitamin-D hormone, 1 alpha,25-dihydroxy-vitamin D(3) [1 alpha,25-(OH)(2)D(3); calcitriol], have emerged that can mimic its various actions in classic calcium transport systems and/or in the regulation of cell proliferation and cell differentiation. Though some of these analogues have accentuated activity in cell differentiation assays in vitro, they lack appreciable "calcemic" activity in vivo, leading to the name "noncalcemic analogues." Several of these analogues are promising candidates for use in treatment of psoriasis and in tumor suppression, one of them, calcipotriol, being already widely approved for the former indication. New generations of calcemic analogues with altered pharmacokinetics are appearing for use in secondary hyperparathyroidism and osteoporosis. We believe that the selective properties of both types of analogues stem from altered receptor binding, blood protein binding, and rate of catabolism. PMID- 18407174 TI - Response. PMID- 18407175 TI - Gastrointestinal peptides and neoplasia ectopic gastric inhibitory polypeptide (GIP) receptors in adrenal glands causing food-dependent Cushing's syndrome. PMID- 18407176 TI - Receptors for secretin, calcitonin, parathyroid hormone (PTH)/PTH-related peptide, vasoactive intestinal peptide, glucagonlike peptide 1, growth hormone releasing hormone, and glucagon belong to a newly discovered G-protein-linked receptor family. AB - Seven receptors with highly homologous structural features have recently been discovered that belong to a new family of seven membrane-spanning receptors within the G-protein-linked receptor superfamily. These seven all bind small peptide ligands, and many have the unique property to activate the G(S) and at least one other G protein. PMID- 18407177 TI - Endocrine manifestations of AIDS. AB - The endocrine manifestations of AIDS are now appreciated to involve virtually any of the endocrine organs. Some of these manifestations are subtle and clinically not apparent. Others, like adrenal involvement, can eventuate, although rarely, into frank glandular insufficiency. This Endocrine Rounds case illustrates a number of endocrine manifestations of AIDS and discusses their involvement from clinical and pathophysiologic aspects. PMID- 18407178 TI - Methods of body composition analysis the status of the gold standard. AB - The prevention, treatment, and comprehension of disease processes can be advanced with improved methods for assessing changes in body composition. Recently developed methodology has expanded our capacity to quantify body constituents and has stimulated a trend away from the two-compartment model (fat mass and fat-free mass) for describing body composition. As these advances take place, the "gold standard" for measuring living human subjects will likely shift from the frequently relied upon underwater weighing method to techniques that further divide the fat-free mass. New and widely used techniques for quantifying various body compartments are described. PMID- 18407179 TI - Cytoplasmic-nuclear trafficking of steroid hormone receptors. AB - The nuclear localization of most steroid hormone receptors reflects a dynamic process: the receptor constantly diffuses out of the nucleus and is reimported by an active mechanism. The outward movement from the nucleus of the receptors and of other nuclear proteins is also mediated by the nuclear localization signals. PMID- 18407180 TI - The SRY gene. AB - SRY was identified in an attempt to clone the mammalian testisdetermining factor. Evidence in the form of sex-reversing mutations and mouse transgenesis experiments indicates that SRY is the testis-determining factor. The cloning of this gene marks the end of the beginning of our understanding of sex determination. This review covers the structure of the SRY gene and its RNA transcripts, along with evidence equating SRY with the mammalian testis determining factor, its evolution, and biochemical aspects of the SRY protein. PMID- 18407181 TI - Neurosteroids Biosynthesis and function. AB - The term neurosteroids applies to those steroids that are both synthesized in the nervous system, either de novo from cholesterol or from steroid hormone precursors, and that accumulate in the nervous system to levels that are at least in part independent of steroidogenic gland secretion rates. Glial cells play a major role in neurosteroid formation and metabolism. Several neurosteroids are involved in either auto- or paracrine mechanisms involving both regulation of target gene expression and effects on membrane receptors (including those of neurotransmitters). The neuromodulatory role of neurosteroids in regulating the estrous cycle and pregnancy, stress, memory, and developmental as well as aging processes awaits further investigation. PMID- 18407182 TI - Multimeric cytokine receptors. AB - Cytokines are pleiotropic regulators that control proliferation, differentiation, and other cellular functions. They play a pivotal role in the immune and hematopoietic systems, and subsets of cytokines often exhibit similar biologic activities in common target cells. Recent cloning of genes for the cytokine receptors has provided new insights into the understanding of how the cytokines control the immune and hematopoietic systems. Cytokine receptors usually consist of multiple subunits, most of which belong to a new gene family called the cytokine receptor superfamily, and receptors for a subset of cytokines with similar functions share a common subunit with a signaling function, providing a molecular basis for the functional redundancy of cytokines. PMID- 18407183 TI - Molecular biology of amphibian metamorphosis A new approach to an old problem. AB - Amphibian metamorphosis has attracted the interest of developmental biologists for decades. Its dependency on thyroid hormone (TH) makes it a unique system to study postembryonic organ development as well as hormone action. Recent cloning o f the TH receptors and the demonstration that these receptors are transcription factors suggest that TH controls metamorphosis by regulating a cascade of gene expression. Systematic analyses of TH early-response genes in three very different organs (the tail, limb, and intestine) have been conducted. The developmental profiles of their expression and their identities suggest that these genes play important roles during metamorphosis, for example, controlling the expression of intermediate and/or TH lateresponse genes. In addition, the gene-regulation program that governs amphibian metamorphosis as revealed by these and related studies appears to resemble that induced by the hormone ecdysone during insect metamorphosis. Further investigation on how the hormone regulates early-response genes and how the products of these genes, in turn, participate in tissue remodeling will provide a better understanding of the molecular basis of the hormonal control of metamorphosis. PMID- 18407184 TI - Animal models of Cushing's disease. AB - Cushing's disease, defined as hyperadrenocorticism resulting from excessive secretion of pituitary ACTH, occurs spontaneously and quite commonly in dogs and horses. In dogs, as in humans, the disease is usually associated with a small tumor of the pituitary pars distalis. However, the disease may arise occasionally (dogs) or exclusively (horses) from tumors or hyperplasia of the pituitary pars intermedia. In dogs, pars intermedia tumors may arise from one of two proopiomelanocortin-containing cell types that are present in normal tissue. PMID- 18407185 TI - Structural analysis and regulation of angiotensin II receptors. AB - Angiotensin (Ang II) is an octapeptide hormone that plays a crucial role in the maintenance of electrolyte homeostasis and cardiovascular function. The hemodynamic and cardiovascular effects o f Ang II are mediated by high-affinity cell-surface receptors of the AT(1) pharmacologic class. The mammalian AT(1) receptor has recently been cloned and found to encode a 359-amino-acid protein of 41,000 molecular weight. The AT, receptor belongs to the guanine nucleotide regulatory-proteincoupled receptor family and is coupled to the phospholipase C signal transduction pathway as evidenced by intracellular calcium mobilization and inositol trisphosphate production upon receptor activation. Cloning of the AT(1) receptor has facilitated the study of structure-function correlates and molecular mechanisms of receptor regulation, and will lead to substantial progress in elucidating the mechanisms governing Ang II actions. PMID- 18407187 TI - Spontaneous remission of pheochromocytoma. PMID- 18407186 TI - Iodothyronine deiodinase. AB - The iodothyronine deiodinases constitute a family of enzymes that catalyze the removal of iodine atoms from various thyroid hormones (THs) in the thyroid gland and extrathyroidal tissues. As such, they are responsible for both the activation and inactivation of these compounds, and are thus important regulators of TH action. Recently, new insights have been gained into the biochemical characteristics of these proteins and their physiologic roles in TH metabolism. In particular, the availability of affinity-labeling techniques, molecular probes, and specific antisera for these enzymes, and the recent identification of the type I deiodinase as a selenoprotein, have ushered in a new era in the study of thyroid hormone deiodination. PMID- 18407188 TI - Mechanisms of cellular information processing. AB - Living cells in multicellular organisms are in simultaneous contact with many regulatory factors such as hormones or neurotransmitters. Many of these factors vary with time in their local concentrations, owing to pulsatile release or production. Therefore, complex patterns of signaling factors act on each living cell in vivo, stimulating or inhibiting second-messenger pathways with potentially complex dynamics. These intracellular pathways do not operate independently but are extensively interconnected, creating complex networks and patterns of intracellular signals that combine to determine the cell's response. The potential significance of cross-signaling between second-messenger pathways and of dynamic stimulation of receptors for cellular information processing in physiology and pathophysiology are discussed. PMID- 18407189 TI - Islet cell growth and the growth factors involved. AB - Throughout development, growth and aging, the mass of the pancreatic islets, in particular the insulin producing beta cell, increases to meet the functional demand and maintain euglycemia. Islet growth occurs by two pathways: (a) the expansion by replication of preexisting beta cells and (b) the formation of new islets (neogenesis) by proliferation and subsequent differentiation of pancreatic ductal epithelium. Some of the factors involved in these pathways of islet growth have been defined. PMID- 18407190 TI - New advances in understanding the molecular mechanisms of thyroid hormone action. AB - Thyroid hormone regulation o f gene transcription is a complex process. There are multiple thyroid hormone receptors (TRs) encoded on separate genes that bind to thyroid hormone-response elements (TREs) of target genes containing different orientation and spacing of half-sites. Additionally, there are multiple TR complexes-monomers, homodimers, and heterodimers with other related nuclear proteins-which bind to TREs and may play important roles in gene transcription. Recently, it has been shown that DNA binding of these TR complexes can be differentially regulated by either ligand or TR phosphorylation. Diversity among TR complexes and TREs, as well as mechanisms for regulating TR binding to TREs, may enable sensitive and precise transcriptional control of target genes. PMID- 18407191 TI - Growth Hormone neuroregulation in diabetes mellitus. AB - GH secretion is markedly altered in diabetic rats and humans. Diabetes in the rat, whether occurring spontaneously or after streptozotocin administration, results in depressed GH secretion. This defect is likely caused by an increase in hypothalamic somatostatin tone and decreased pituitary GH. The effects of diabetes in humans depend upon the etiology of the disease. In type-1 diabetes, GH secretion is increased and in type 2 it is decreased. Again, these changes are hypothesized to be due to opposite alterations in hypothalamic somatostatin. Current evidence suggests that GH hypersecretion in human type-1 diabetes may be relevant to important metabolic and angiopathic complications of the disease. PMID- 18407192 TI - Subclinical thyroid dysfunction in the elderly. AB - Subclinical thyroid dysfunction is more common in older persons. By definition, these disorders are recognized by isolated elevation or suppression of the serum TSH concentration, in association with a normal serum free thyroxine level. Among individuals over 65 years old, subclinical hypothyroidism is found in approximately 10% of women and approximately 3% of men. It is most commonly due to autoimmune thyroiditis or previous treatment for hyperthyroidism. There may be three indications for L-thyroxine therapy: (a) presence of antithyroid antibodies, indicating substantial risk of progression to over hypothyroidism; (b) symptoms consistent with thyroid hormone deficiency; and (c) an elevated serum LDL-cholesterol. Subclinical hyperthyroidism is present in approximately 1% 2% of older persons. The most common cause is excessive thyroid hormone therapy, followed by mild endogenous hyperthyroidism due to Graves' disease or nodular goiter. These can be differentiated from other causes of low serum TSH concentration based on clinical and other laboratory and radionuclide scan criteria. The most serious consequences of subclinical hyperthyroidism are atrial fibrillation and osteoporosis, to which elderly patients are particularly predisposed. PMID- 18407193 TI - Control of the preovulatory Luteinizing Hormone surge by Gonadotropin-Releasing Hormone antagonists: prospects for clinical application. AB - The preovulatory LH surge of the primate menstrual cycle represents a number of positive influences, a major component of which is a direct action of estradiol on the anterior pituitary lobe. Whether the LH surge also requires a corresponding burst of GnRH release from the hypothalamus has been debated. After many years of investigation, there is now conclusive evidence that a midcycle GnRH surge does occur in the primate. This is supported by studies in women with normal ovulatory cycles that demonstrate that blockade of the GnRH receptor by potent GnRH antagonists administered within 1-2 days of the expected midcycle can delay the LH surge. The ability to prevent the positive feedback effects of estradiol by GnRH antagonists is being employed for the controlled induction of follicular development and ovulation in the treatment of infertility and in in vitro fertilization programs. PMID- 18407194 TI - Renin-Independent hypermineralocorticoidism. AB - Although renin-independent hypermineralocorticoidism is an uncommon form of hypertension, its diagnosis provides the clinician with a unique opportunity in the field of hypertension, that is, to render a surgical cure or to achieve a dramatic pharmacologic response in the treatment of hypertension. Primary aldosteronism is the most common form of renin-independent hypermineralocorticoidism. The plasma aldosterone concentration to plasma renin ratio is an excellent screening test for primary aldosteronism, the diagnosis of which should be confirmed by demonstrating unsuppressible urine or plasma levels of aldosterone. The subtype of primary aldosteronism dictates the most appropriate therapy. Computerized imaging of the adrenal glands and adrenal venous sampling assist in distinguishing unilateral (requiring surgical treatment) from bilateral (requiring pharmacologic treatment) adrenal disease. The forms of mineralocorticoid excess considered in the hypokalemic hypertensive patient with low aldosterone values include congenital adrenal hyperplasia (11 beta-hydroxylase and 17alpha-hydroxylase deficiencies), deoxycorticosterone producing tumor, Cushing's syndrome, primary cortisol resistance, and 11 beta hydroxysteroid dehydrogenase deficiency (apparent mineralocorticoid excess syndrome). The I1 beta-hydroxysteroid dehydrogenase deficiency may be congenital or acquired (for example, ingestion of licorice or carbenoxolone). PMID- 18407195 TI - Role of the hypophyseal neurointermediate lobe in the dynamic release of prolactin. AB - The dynamic release of prolactin (PRL) occurs in a region of the anterior pituitary lobe (AP) that receives its blood supply primarily from the hypophyseal neurointermediate lobe (NIL) via the short portal vessels. This relationship, coupled with a growing body of supportive evidence, suggests that the hypophysiotropic signal mediating the massive discharge o f AP PRL derives from the NIL rather than the hypothalamic median eminence, as previously believed. Moreover, a major component of this signal appears to be a-MSH. PMID- 18407196 TI - Breast cancer and expression of aromatase in breast adipose tissue. AB - Extraglandular conversion of C19 steroids to estrogens takes place primarily in the stromal cell component of adipose tissue and is catalyzed by an enzyme complex comprising aromatase cytochrome P450 (P450arom; the product of the CYP19 gene) together with the flavoprotein NADPH-cytochrome P450 reductase. It has been proposed that increased local aromatase activity in breast adipose tissue may influence the growth o f breast carcinomas. Using competitive polymerase chain reaction following reverse transcription (RT-PCR), we quantified P450arom transcripts in breast adipose tissue from mastectomy specimens. In 10 of 15 patients, the highest transcript levels were found in the quadrant where the tumor was located. We also found the highest proportions of adipose stromal cells versus adipocytes in these quadrants. These findings suggest that regional differences in the relative proportions o f these histologic components give rise to local elevated concentrations of estrogens. Although the initiating events are not known, once a neoplastic change has occurred, tumor growth may be promoted by these locally increased estrogen levels. Our data regarding hormonally regulated use of tissue-specific promoters for CYP19 gene transcription suggest preferential utilization of certain upstream regulatory regions of the gene in breast adipose tissue, resulting in increased expression of aromatase. PMID- 18407197 TI - Melanin-concentration hormone updated functional considerations. AB - The melanin-concentrating hormone (MCH) is a vertebrate neuropeptide produced in hypothalamic perikarya whose fibers project to most regions of the brain and into the spinal cord. Its role as a neurohypophyseal color-change hormone is peculiar to teleost fish, but recent studies in mammals suggests that MCH itself, and other peptides derived from the same precursor, may participate in multiple functions in the central nervous system, modulating behavior and the perception of sensory information. Recent hybridization studies in mammals have greatly increased our understanding of the response of the MCH system to environmental factors, such as osomotic challenge, lactation, stress, and changes in corticosteroid levels. Further studies in lower vertebrates are needed to highlight the physiologically important functions that have led to the structural conservation of the MCH peptide during vertebrate evolution. PMID- 18407198 TI - Paracrine control of spermatogenesis. AB - Spermatogenesis is a complex process that is both remarkable and enigmatic. While circulating hormones clearly play an important role in initiating and regulating the process, the Sertoli cell barrier prevents most substances from entering the seminiferous tubule compartment and directly influencing germ cell development. Therefore, the testis cannot rely solely upon the delivery of circulating hormones, nutrients, and growth factors, but must independently produce its own regulatory substances. A rapidly increasing number of testicular factors that appear to modulate spermatogenesis in a paracrine fashion are now being identified. These discoveries are beginning to contribute to our understanding of the intricate network of testicular cell-cell interactions that control male reproduction. PMID- 18407199 TI - Hormonal regulation of adipose differentiation. AB - Adipose differentiation is a multistep process with the following sequence: adipoblasts --> preadipocytes --> adipocytes. Adipogenic agents are only involved in the terminal differentiation of preadipocytes to adipocytes by means of circulating hormones (growth hormone, glucocorticoids, or triiodothyronine) and locally produced hormones (prostacyclin). Fatty acids also behave as hormones and act as transcriptional regulators of lipid-related genes. Once differentiated, adipocytes become secretory cells able to synthetize and release an impressive number of peptide and nonpeptide compounds, suggesting a potential link between excess of adipose tissue mass and various physiopathphysiologic consequences. PMID- 18407200 TI - Signaling by the cytokine receptor superfamily just another kinase story. AB - Many cytokines initiate cellular responses through their interaction with members of the cytokine receptor superfamily. This family of receptors contains no catalytic domains in the cytoplasmic domain, but all couple ligand binding to tyrosine phosphorylation, and this activity requires a membrane-proximal region that contains some similarity among the receptors. Recent studies have shown that members of the JAK family of protein tyrosine kinases associate with the membraneproximal region, are rapidly tyrosine-phosphorylated following ligand binding, and their in vitro kinase activity is activated. The JAK family of kinases is characterized by two kinase domains, only one of which contains all of the hallmarks of active kinases. This family of 130-kD kinases lacks SH2 or SH3 domains, but family members contain extensive homology in the large amino terminal region. Individual receptors associate with, or require, one or more of the three known family members including JAKI, JAK2, and tyk2. Putative substrates of the JAK family of kinases include the 91-kD and 113-kD proteins of the interferon-stimulated transcription complex ISGF3 that, when tyrosinephosphorylated, migrate to the nucleus and participate in the activation of gene transcription. Recent evidence suggests that the 91- and 113-kD proteins are members of a large family of genes that are potential substrates o f JAK family members and may regulate a variety o f genes involved in cell growth, differentiation, or function. Together the data provide a new, generalized model for the mechanisms by which cytokines that utilize receptors of the cytokine receptor superfamily regulate cellular activity. PMID- 18407201 TI - Development of the mammary gland and lactation. AB - Many hormones, growth factors, and protooncogene products involved in mammary development and lactation have been identified; however, the mechanism of their concerted action remains to be explained. In addition to these regulatory factors, normal mammary development and lactation require the cell-cell interaction of stromal and parenchymal elements in the mammary gland. Recent studies indicate that PRL effects on target cells are likely transmitted into cells via activation of tyrosine kinase associated with a protein-designated JAK 2, while other studies have identified stimulatory and inhibitory factors that interact with the 5' promotor regions of milk-product genes. PMID- 18407202 TI - Postreceptor signaling mechanisms for Growth Hormone. AB - Recent data have shed significant new light on the mechanisms involved in the transmission of a biologic signal by GH. Following ligand-induced dimerization of the GH receptor, multiple cascades are involved in GH signaling. These include activation of nonreceptor tyrosine kinases, in particular JAK2, which is a mechanism shared by the newly described cytokine receptor superfamily. Furthermore, several classic pathways (for example, guanine-nucleotide-binding proteins and protein kinase C), shared by numerous hormones, growth factors, and neurotransmitters, are also involved in many of the actions of GH. The interrelationships between the various signaling pathways for GH have not yet been fully defined. This review briefly summarizes the current state of knowledge with respect to the processes involved in the effects of GH in target cells. PMID- 18407203 TI - Receptor to nucleus signaling via tyrosine phosphorylation of the p91 transcription factor. AB - Recent studies on signal transduction stimulated by interferons have defined pathways that link cell surface receptors to target genes in the nucleus. After interferon binding, nonreceptor tyrosine kinases are activated that phosphorylate components of latent DNA-binding factors in the cytoplasm. These phosphorylated factors form multimeric complexes that translocate to the nucleus and bind to specific DNA sequences in the promoters of induced genes. A 91-kD factor (p91 or Stat 91) is activated by interferons and serves as a subunit partner in the composition of diverse transcription factors. PMID- 18407204 TI - The Sos (Son of sevenless) protein. AB - The Drosophila Son of sevenless (Sos) gene functions in the signaling pathway initiated by the Sevenless receptor tyrosine kinase. It encodes the Drosophila homologue of CDC25, an activator of Ras in the yeast Saccharomyces cerevisiae. Two widely expressed mammalian homologues of Sos (mSos) have now been identified and characterized. They encode for 150-kD proteins that are Ras-specific guanine nucleotide exchange factors. Genetic and biochemical studies indicate that Sos proteins bind directly to the SH2- and SH3-domain-containing adaptor protein GRB2/Drk. This interaction defines a pathway by which receptor tyrosine kinases can communicate with Ras. PMID- 18407205 TI - Immunosuppressive therapy and the skeleton. AB - Glucocorticoids and other immunosuppressive drugs, such as cytoclosporine A, are increasingly used today. One of the most common clinical situations in which they are prescribed is for immunosuppression after organ transplantation. These drugs have diverse effects on the skeleton, however, and one of the most common sequelae of organ transplantation is osteoporosis and fractures. Patients treated with immunosuppressive drugs should be carefully evaluated for osteoporosis, preferably prior to or at the time of initiation of immunosuppressive therapy. They should be followed carefully with sequential bone-density measurements and biochemical indices of bone turnover. Measures to prevent bone loss should be initiated early in the course of immunosuppressive therapy. PMID- 18407206 TI - The aromatic hydrocarbon receptor, dioxin action, and endocrine homeostasis. AB - The environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin acts through the aromatic hydrocarbon (AH) receptor to enhance CYP1A1 gene transcription. There are notable mechanistic analogies between dioxin action and lipophilic hormone action. In addition, some of dioxin's adverse effects are associated with disruptions in endocrine homeostasis. Thus, dioxin biology intersects several disciplines, including toxicology, endocrinology, molecular biology, and public health. PMID- 18407207 TI - The discovery of 9-cis retinoic acid: a hormone that binds the retinoid-X receptor. AB - Two classes of nuclear receptors, the retinoic acid receptors (RARs) and the retinoid-X receptors (RXRs), mediate the physiologic activity of retinoids. The RXRs can form biologically active heterodimers with the RARs and with other nuclear receptors, including the vitamin-D, thyroid hormone, and peroxisome proliferator-activated receptors. Thus, the RXRs may play a pivotal role in modulating the action of other hormones or ligands. The RXRs were originally classified as orphan receptors whose cognate ligand was unknown until recently, when 9-cis retinoic acid (9-cis RA) was discovered to bind directly and activate this family of receptors. Since 9-cis RA also binds and activates the RARs, it is interesting to speculate that this natural ligand may regulate a broad range of physiologic processes by mediating transcriptional activity through both RAR- and RXR-linked pathways. PMID- 18407208 TI - Phosphorylated and nonphosphorylated Prolactin isoforms. AB - Specific posttranslational modifications o f monomer PRL produce a number of hormone variants. This review describes the current state of knowledge for the nonphosphorylated and phosphorylated 24-kD monomer forms. Of particular interest and significance for our future understanding of PRL endocrinology are the studies showing their differential release in response to different physiologic signals and their different activities in target tissues. In at least two target tissues, the monophosphorylated variant seems to act as an antagonist to the unmodified hormone. PMID- 18407209 TI - Gonadotropin-releasing hormone neurons: intrinsic pulsatility and receptor mediated regulation. AB - The pulsatile pattern of gonadotropin-releasing hormone (GnRH) release from the hypothalamus is driven by a functionally interconnected and synchronized network of GnRH neurons termed the GnRH pulse generator. Several recent observations have revealed that immortalized GnRH neurons can generate an episodic pattern of GnRH release when cultured in the absence of other cell types. The in vitro operation of the pulse generator depends on the development of synaptic contacts among GnRH neurons, the electrical properties of individual GnRH neurons, and the GnRH induced modulation of its secretory mechanism. The expression o f several other receptors by GnRH neurons provides the means for integrated regulation of pulse generator activity from without the network by agonists including glutamate, GABA, endothelin, and catecholamines. PMID- 18407210 TI - Molecular insights into inherited ACTH resistance syndromes. AB - The ACTH resistance syndromes-familial glucocorticoid deficiency (FGD) and the triple-A syndrome-have long been postulated to result from genetic defects of the ACTH receptor. We have demonstrated a point mutation that reduced function of this receptor in FGD, and subsequently we, and others, have identified other mutations of this gene in other families with this condition. Gene linkage studies, however, show that the ACTH receptor is not associated with either a subgroup o f FGD in which mutations in the ACTH receptor gene cannot be found or with the triple-A syndrome. The study of these diseases may reveal new aspects of adrenal development and function, and provide insights into the molecular mechanisms of ACTH receptor action. PMID- 18407211 TI - Role and site of estrogen action in follicular atresia. AB - Subsequent to an initial understanding that estrogen was only stimulatory to folliculogenesis, we have come full circle to the present recognition that many actions of estrogen are inhibitory to follicular function. The development of this interpretation has frequently been associated with the controversial issue o f the likely site o f estrogen action, especially in primates, where much of the evidence has been amassed. The accumulated findings in a variety of species seem to demonstrate clearly that at least part of the atretogenic effect of estrogen is exerted directly on the ovary, apparently by interaction with the nuclear estrogen receptor. Recent observations include identification of messenger RNAs for the estrogen receptor and for creatine kinase in the macaque ovary, and a current focus is to localize messages within specific compartments of the ovary. PMID- 18407212 TI - Ion channels and the signal transduction pathways in the regulation of growth hormone secretion. AB - The secretion of GH from pituitary somatotrophs is mainly regulated by alterations in the levels of intracellular free Ca(2+) concentrations ([Ca(2+)](i)) that depend on the influx of Ca(2+) through voltage-gated Ca(2+) channels in the cell membrane. Hypothalamic stimulatory and inhibitory factors bind to specific receptors on the cell membrane to regulate membrane potential and activate second-messenger systems. The receptors are G-protein coupled, and activated G proteins directly influence membrane ion channels to regulate Ca(2+) influx. The function of cAMP-dependent protein kinase A is also modulated by receptor-coupled G proteins leading to the phosphorylation of Ca(2+) channel proteins and further alteration of Ca(2+) influx. PMID- 18407213 TI - COUP-TF an orphan member of the steroid/thyroid hormone receptor superfamily. AB - COUP-TFs are orphan members of the steroid/thyroid hormone receptor superfamily. They are transcription factors that are highly conserved across species and have a wide spectrum of binding specificity. In tissue cultures, COUP-TFs repress the transactivation function of several other members of the steroid/thyroid hormone receptor superfamily, and they also negatively regulate the transcription from promoters of many other genes. The expression profile of COUP-TFs in several species suggests that they play an important role in development and differentiation. PMID- 18407214 TI - The role of the aryl hydrocarbon receptor nuclear translocator protein in aryl hydrocarbon receptor action. AB - The aryl hydrocarbon (AH, or dioxin) receptor mediates carcinogenesis by a wide variety of compounds. It acts as a ligand-dependent transcription factor. Many investigators expected that the AH receptor would prove to be a member of the steroid/thyroid/retinoic acid receptor superfamily of proteins. However, recent cloning of the two subunits of the DNA-binding form of the AH receptor has shown that this is not the case. These subunits, the AH receptor nuclear translocator protein (ARNT) and the ligand-binding AH receptor monomer, do not contain zinc finger DNA-binding domains, nor do they have any other sequence similarities with members of the above family. Instead, they both contain basic helix-loop-helix (bHLH) motifs and also another segment of sequence similarity, the "PAS" region. bHLH motifs in other transcription factors are known to function as dimerization and DNA-binding domains. Present experiments are directed toward understanding the mechanisms of action and the roles of the two subunits. PMID- 18407215 TI - Mutations of the low-density-lipoprotein receptor gene and familial hypercholesterolemia. AB - Familial hypercholesterolemia (FH), an autosomal dominant disorder caused by mutation of the low-density-lipoprotein (LDL) receptor, occurs in about one in 500 individuals. The evaluation of naturally occurring mutants has permitted an extensive structure-function analysis of this receptor that has provided insight into the biochemistry and cell biology of cell-surface receptors in general. Novel gene therapeutic approaches to the management of FH are a developing outgrowth of this research. PMID- 18407216 TI - Phosphoinositides and calcium signaling New aspects and diverse functions in cell regulation. AB - Numerous circulating and locally produced hormones bind to specific cell-surface receptors and activate a variety of second-messenger pathways that evoke characteristic phenotypic responses in their target cells. One of the most ubiquitous signal transduction mechanisms is the phosphoinositide-calcium messenger system, which is activated by hormones, neurotransmitters, and growth factors. Stimulation of these receptors by their ligands causes a characteristic change in the metabolism of membrane phospholipids with production of diacylglycerol and a rapid increase in cytoplasmic Ca(2+) concentration, due to the release of stored intracellular Ca(2+) and stimulated Ca(2+) entry from the extracellular space. These intracettular signals act in concert to activate protein kinases that phosphorylate a variety of regulatory proteins. The link between phosphoinositide turnover and Ca(2+) mobilization is inositol 1,4,5 trisphosphate, the major Ca(2+)-mobilizing second messenger, which is produced from membrane phosphoinositides by activated phospholipase C enzymes. The mechanisms of ligand-regulated Ca(2+) influx and the additional regulatory role(s) of phosphoinositides and inositol phosphates are still being unfolded. This review and the following article summarize some recent developments and unsolved issues about this major signal transduction cascade that links calcium mobilizing hormone receptors to the regulation of endocrine cell function. PMID- 18407217 TI - Calcium mobilization by inositol phosphates and other intracellular messengers. AB - Inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] is now widely recognized as a messenger controlling the release of calcium from intracellular stores. In oocytes, and also probably in excitable cells, another potential calcium mobilizing messenger is cyclic ADP ribose, although there is as yet little evidence that its levels are regulated by hormones or other extracellular mediators. In addition to signaling intracellular calcium release, [Ins(1,4,5)P(3)] also regulates calcium entry across the plasma membrane, but not in a direct manner. Rather, the depletion of intracellular stores by the calcium mobilizing action of [Ins (1,4,5)P(3)] initiates a process of retrograde signaling whereby the depleted stores generate or release a diffusible messenger that is believed to act on the plasma membrane. A phosphorylated metabolite of [Ins(1,4,5)P(3)], inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P(4)], has been proposed to modulate this process, but the literature is not consistent on this point. A recently proposed candidate for the retrograde messenger is an activity extracted from Jurkat cells termed CIF (calcium influx factor), which has many properties consistent with such a messenger. There is also evidence that a GTP dependent process, possibly involving a small G protein, is involved in signaling calcium entry and may be involved in either the formation or action of the diffusible messenger for calcium entry. PMID- 18407218 TI - Role of the Rab3A GTPase in regulated secretion from neuroendocrine cells. AB - Regulated secretion is a complex process that involves scores of distinct proteins. These proteins must assemble into machines that organize vesicle docking, membrane fusion, and recovery of vesicle components. Rab3A, a member of the Ras superfamily of GTPases, associates with secretory granule membranes of neuroendocrine cells, and mutants of Rab3A can inhibit exocytosis. A model is proposed in which Rab3A controls the assembly and disassembly of a docking complex that inhibits membrane fusion until an external signal triggers release of the Rab3A from the complex. PMID- 18407219 TI - Corticotropin-release inhibitory factor Evidence for dual stimulatory and inhibitory hypothalamic regulation over adrenocorticotropin secretion and biosynthesis. AB - The hypothalamus is currently thought to stimulate the synthesis and secretion of ACTH uniquely by secreting neuropeptides into the hypophysial-portal circulation, of which the most important are CRH and arginine vasopressin. However, analysis of the effects of pituitary isolation on the pituitary-adrenal axis in a variety of species suggests that the hypothalamus exerts both stimulatory and inhibitory regulation over ACTH secretion and POMC biosynthesis, and that the inhibitory control is dominant. Because none of the currently known inhibitory factors in the hypophysial-portal circulation consistently decreases basal ACTH secretion and POMC mRNA levels in normal anterior pituitary cells, it is suggested that this inhibition is mediated by a currently unidentified hypothalamic substance, presumably a neuropeptide, which we have termed corticotropin-release inhibitory factor (CRIF). The possible roles in clinical medicine of agonists and antagonists of this putative CRIF are discussed. PMID- 18407220 TI - Autoregulation and crossregulation of nuclear receptor genes. AB - Whereas crossregulation of nuclear receptors has been known for some time, recently several examples of autoregulation have been described, especially during development and specific gene expression. In this review, I discuss both these phenomena, based on some studies from our laboratory on amphibian metamorphosis and egg protein gene expression. These include autoinduction of estrogen receptor (ER) accompanying egg protein gene expression in adult and larval Xenopus; autoinduction of thyroid hormone receptor (TR) during metamorphosis and in adult Xenopus; crossregulation by triiodothyronine (T(3)) and dexamethasone of autoinduction of ER; and inhibition by PRL of autoinduction and crossinduction of TR and ER genes. A dual receptor threshold model to explain the interplay between T(3), estrogen and PRL is presented and its significance to the general question of nuclear receptor autoregulation and crossregulation during development is also discussed. PMID- 18407221 TI - Gonadal steroid hormone regulation of the somatotropic axis during puberty in humans Mechanisms of androgen and estrogen action. AB - The adolescent growth spurt is associated with a sex steroid hormone-dependent rise in GH production; both androgens and estrogens are implicated as positive regulators of the somatotropic axis during puberty. The issue is complicated by the fact that testosterone may act both directly via the androgen receptor and indirectly, after its aromatization to 17beta-estradiol, through the estrogen receptor. Recently, a number of investigators have studied the effects of the administration of androgen and estrogen receptor antagonists, as well as nonaromatizable androgens, on GH secretion. These reports suggest that estrogen receptor-dependent processes play a facilitatory role in the pubertyassociated rise in GH secretion. If androgen receptor-mediated events are involved in the control of the somatotropic axis, their role is likely inhibitory. A hypothalamic site of action of the sex steroids is postulated. PMID- 18407222 TI - Growth in growth hormone insensitivity. AB - Primary GH insensitivity due to GH receptor deficiency (GHRD) provides a model for studying the discrete effects of severe IGF-I deficiency on growth and body composition. Growth failure in utero is doubtful, but postpartum growth proceeds at rates that result in adult statures 4-12 standard deviations (SDs) below the normal mean. Wide variability in statural effect, even in a genetically homogeneous population, is partly explained by correlation of SD score with biochemical measures of GH effect (IGF-I, IGF-II, and IGFBP-3). Growth and changes in body composition (decreased fat/lean) in patients with GHRD in response to exogenous IGF-I indicate that direct local effects of GH are not necessary for these responses. PMID- 18407223 TI - Neuroendocrine-immune system interactions The immune-hypothalamo-pituitary adrenal axis. AB - Bidirectional interactions between the immune system and the hypothalamo pituitary-adrenal (HPA) axis may represent an important mechanism that prevents overexuberant immune responses. In this network, cytokines play an important role in modulating the HPA axis response at all three levels: the hypothalamus, the pituitary gland, and the adrenals. Acute effects of cytokines are produced at the hypothalamic level by altered release of hypophysiotropic hormones, whereas pituitary and adrenal actions are slower but probably more important in response to prolonged release of cytokines, as it occurs in infection or inflammation. Further studies are needed to determine the precise mechanism through which circulating cytokines affect the HPA axis function within the central nervous system and to evaluate the relevance of these interactions in physiologic and pathophysiologic situations in humans. PMID- 18407224 TI - Developmental regulation of the ovary via growth factor tyrosine kinase receptors. AB - Ovarian development and mature ovarian function are subjected to the regulatory influence of several growth factors exerting their biologic actions via membrane anchored receptors endowed with intrinsic tyrosine kinase activity. The actions of these growth factors appear to be anatomically, functionally, and temporally coordinated. Whereas transforming growth factor alpha (TGFalpha)-produced in thecal cells- facilitates cell proliferation and slows gonadotropin-dependent biochemical differentiation of both thecal and granulosa cells, IGF-I-produced in granulosa cells-amplifies the effect of gonadotropins on this ovarian compartment. The interactive paracrine-autocrine influence of both growth factors may facilitate the simultaneous occurrence of the two basic processes underlying follicular development: growth and cytochemical differentiation. Upon completion of preovulatory development, fibroblast growth factors (FGFs)-produced by granulosa, and also perhaps by thecal-interstitial cells-may come into play, accelerating cytodifferentiation of thecal cells into luteal cells by reducing the androgenic response of thecal cells to gonadotropins. In addition, FGFs may facilitate proliferative events underlying the organization of pregranulosa cells during early ovarian development. Neurotrophins may exert their greatest impact during these same developmental windows affected by FGF. Neurotrophins may contribute to regulating cell-cell interactive processes related to follicular organization and/or formation during early ovarian development, and to the preovulatory cytodifferentiation of thecal-interstitial cells that precedes ovulatory rupture. Neurotrophins may also regulate the density of follicular innervation. Because each of these growth factors binds to a different receptor tyrosine kinase and expression of each receptor is highly compartmentalized and developmentally regulated, it appears clear that activation of receptor tyrosine kinases is intimately linked to both normal and deranged ovarian development. PMID- 18407225 TI - Sepsis and adrenal function. AB - In the setting of sepsis, adrenal function can be difficult to evaluate. Cortisol levels, normally elevated by the stress of sepsis, are occasionally reduced, signifying possible adrenal dysfunction. Even elevated cortisol levels do not assure that adrenal reserve is adequate and may in fact portend a preterminal state. Bilateral adrenal hemorrhage leading to adrenal insufficiency is one complication of the sepsis syndrome. This endocrine rounds illustrates the importance in considering adrenal insufficiency and adrenal hemorrhage in patients with overwhelming sepsis while discussing the pathophysiology, clinical presentation, and therapeutic implications of this dire complication. PMID- 18407226 TI - The role of oxidation and glycation in the pathogenesis of diabetic atherosclerosis. AB - Diabetes carries an increased risk of atherosclerotic disease that is not fully explained by known cardiovascular risk factors. There is accumulating evidence that advanced glycation of structural proteins, and oxidation and glycation of circulating lipoproteins, are implicated in the pathogenesis of diabetic atherosclerosis. Antioxidants can inhibit oxidation and advanced glycation in vitro, and antioxidant therapy can prevent cardiovascular disease in nondiabetic individuals. Large-scale clinical studies are required to determine the therapeutic role of antioxidants and specific antiglycation agents in diabetic vascular disease. PMID- 18407227 TI - Use of Growth Hormone in children with short stature and normal Growth Hormone secretion: a growing problem. AB - The ethical, economic, psychologic, social, and growth attainment outcome issues related to the use of GH therapy in normal children with short stature are discussed. Although some short children accelerate their growth velocity with GH treatment, the limited available data do not suggest a significant benefit in final height attainment. An international survey of 99 normal short children treated with GH for at least 3 years reported a net improvement in mean height gain of <1 cm/year. Only in one-third was the result considered very good or good; 40% stated that there was no benefit. Thus, it seems unlikely that GH will dramatically increase final height in short children. On this basis, the use of GH for the treatment of the normal child with short stature outside of carefully controlled clinical trials cannot be recommended at present. PMID- 18407228 TI - Mutual cross-modulation of steroid/retinoic acid receptor and AP-1 transcription factor activities: a novel property with practical implications. AB - Nuclear receptors and the AP-1 transcription factors (homodimers and heterodimers of members of the Jun/Fos/ATF/families) mutually influence their activities, positively and negatively. Mutual negative interference is of practical importance as it can be exploited for therapy of human diseases. Since the discovery of cross-modulation, many reports, seemingly not always compatible, have appeared. Recent experimental evidence may help to develop a model for this interesting transcription factor interaction that satisfies most observations. Transactivation and transrepression (as one facet o f cross-modulation) are separable activities o f the nuclear receptors. The modulating property appears to be exerted by nuclear receptor monomers. These conclusions are supported by point mutations in the glucocorticoid receptor and by the specific inhibitory effects of ligand analogues on transactivation only. These pilot experiments make a search for ligands that enhance the cross-modulatory function without triggering transactivation very promising. PMID- 18407229 TI - Nongenomic actions of steroid hormones. AB - The genomic theory of steroid action has dominated research in the field over the past 4 decades. Despite early observations on rapid steroid effects being clearly incompatible with this theory, only recently has immediate steroid action been more widely recognized and is summarized here, with the emphasis on aldosterone. Further evidence for nongenomic steroid effects is also gathered rapidly for various groups of other steroids, such as neurosteroids, vitamin D(3), and estrogens. PMID- 18407230 TI - The human genome project: genetic and physical mapping. AB - The modern tools of molecular biology, recombinant DNA techniques, have given scientists the ability to isolate and study individual genes from even complex eukaryotic genomes. The availability of genes enables the study o f their structure and biologic function, and their role in normal and abnormal physiologic processes. A worldwide effort to study and understand the entire human genome is under way, which will result in information on the location of all genes, their sequences, and their complex regulation and interactions. As this knowledge becomes available, it will be rapidly applied to the practice of medicine through use in the development of diagnostic tests for genetic-based diseases and in the development of therapeutics. PMID- 18407231 TI - Current evaluation of thyroid nodules. AB - Thyroid nodules are common and frequently benign. Current evaluation by fine needle aspiration (FNA) biopsy is efficacious and cost-effective. Results of FNA series confirm that this procedure is safe, without serious complications, reliable, and relatively easy to perform. Radioisotope scanning and high resolution ultrasonography are used in special circumstances and not as routine diagnostic tests. When FNA biopsy is applied, benign nodules (75%) are followed medically, and suspicious (20%) or malignant (5%) nodules are excised surgically. Overall, FNA biopsy of thyroid nodules has improved surgical selection, resulting in a 25% reduction in thyroidectomies for benign nodules. PMID- 18407232 TI - Insulin signal transduction pathways. AB - Insulin initiates its pleiotropic effects by activating the insulin receptor tyrosine kinase to phosphorylate several intracellular proteins. Recent studies have demonstrated that phosphotyrosine residues bind specifically to proteins that contain src homology 2 (SH2) domains, and that this interaction mediates the regulation of multiple intracellular signaling pathways. This article reviews recent progress in elucidating the detailed pathways that lead from the insulin receptor to the ultimate biologic actions of insulin. PMID- 18407233 TI - Multifunctional role of nitric oxide in inflammation. AB - In response to infection or tissue damage, an array of soluble and lipid mediators as well as cytokines and growth factors cause both immune and nonimmune cells to produce rather large amounts of nitric oxide. Nitric oxide and its oxidation products are toxic and can cause tissue injury. The endocrine system can protect against nitric-oxide-mediated tissue damage by producing corticosteroids, growth factors, and cytokines that are potent inhibitors of nitric oxide production. This review focuses on our current understanding of the role of nitric oxide in the inflammatory response. An emphasis has been placed on the potential for nitric oxide in tissue damage. PMID- 18407234 TI - Activation of tyrosine kinase signal pathways by radiation and oxidative stress. AB - Most research on ionizing radiation, ultraviolet radiation, and H2O2 exposure has focused on the well-known ability of such agents to damage cellular components, particularly DNA. However, recent studies have shown that these events also act directly on components of tyrosine kinase signal transduction pathways, resulting in their activation. Cells use these types of pathways to transmit signals from surface receptors to the nucleus in response to a wide variety of stimuli, ranging from hormones and growth factors such as insulin, erythropoietin, and epidermal growth factor to antigen stimulation of lymphocytes. We propose that cellular responses to radiation and oxidative stress involve the active process of tyrosine kinase signal transduction, in addition to damage to DNA and other cellular components, leading to the activation of transcription factors and the subsequent induction of gene expression. The ability of radiation and oxidative stress to bypass control by normal ligands to act on receptors and their signal transduction pathways offers a new perspective on the ways in which organisms can respond to stress. PMID- 18407235 TI - Signal transduction by calcitonin Multiple ligands, receptors, and signaling pathways. AB - Calcitonin (CT) is a peptide hormone that is secreted by the parafollicular cells of the thyroid in response to elevated serum calcium levels. It acts to reduce serum calcium by inhibiting bone resorption and promoting renal calcium excretion. In addition to this hypocalcemie effect, calcitonin modulates the renal transport of water and several ions other than calcium and acts on the central nervous system to induce analgesia, anorexia, and gastric secretion. The CT receptor, a member of a newly described family of serpentine G protein-coupled receptors, has recently been shown to couple to multiple trimeric G proteins, thereby activating several signaling proteins, including protein kinase C, cAMP dependent protein kinase and calcium/calmodulin-dependent protein kinase. In kidney proximal tubule cells (LLC-PK1), the CT-activated signaling mechanisms vary in a cell cycle-dependent manner, with the receptor coupling through a G(s) protein during G(2) phase and through a G(i) protein and possibly a G(q) protein during S phase. These signaling mechanisms differentially modulate the activities of Na(+)/K(+)-ATPase and the apical Na(+)/H(+) exchanger, effector molecules that play important roles in transepithelial Na(+) transport. Cloning of CT receptors has revealed the presence of alternatively spliced cassettes, resulting in the expression of different isoforms of the receptor. The availability of these recombinant CT receptors has allowed preliminary characterization of the effects of changes in the receptor's structure on its ligand binding and signal transduction properties. Thus, the cellular and molecular biology of CT is complex, with several structurally related peptide ligands and multiple isoforms of the CT receptor that can independently activate diverse signaling pathways. As the recent exciting results in this field are extended, we can expect rapid progress in understanding the molecular basis of the diverse effects of CT and, possibly, of the CT-related peptides CGRP and amylin. PMID- 18407236 TI - Regulation of phospholipase C-beta isozymes by G-proteins. AB - Phospholipase C (PLC) isozymes are known to be regulated, in part, by heterotrimeric GTP-binding protein (G-protein) subunits, including Galpha subunits of the G(q) family and Gbetagamma subunits. New data show that PLC can also be regulated by a high molecular weight G-protein that doubles as a cellular transglutaminase. Furthermore, a soluble phosphatidylinositol transfer protein (PITP) has been implicated in sustaining the activity of PLC by delivering substrate to the plasma membrane. Such diverse regulatory mechanisms imply that the PLC isozymes are precisely controlled and have specific roles in generating second messengers in response to various external stimuli. PMID- 18407237 TI - Inhibin and activin Molecular aspects of regulation and function. AB - Inhibins and activins exhibit a broad spectrum of biologic activities, many of which affect the reproductive axis. Within the ovary and testis, the synthesis of the inhibin and activin alpha and beta subunits is regulated by circulating hormones, such as the pituitary gonadotropins, and by diverse paracrine factors. Considerable progress has been made in establishing patterns of inhibin and activin subunit gene expression in the gonads and in cultured gonadal cells. Analysis of the inhibin and activin subunit genes is now providing insight into the signaling pathways and molecular mechanisms by which inhibin and activin subunit gene expression is modulated in the ovary and testis. Genetic manipulation of the inhibin and activin subunit genes promises additional revelations on the biologic functions of these intriguing hormones. PMID- 18407238 TI - X-linked muscular atrophy and the androgen receptor. AB - X-linked muscular atrophy is a form of adult-onset, usually slowly progressive spinal and bulbar motor neuron degenerative disease that is uniquely associated with male hypogonadism. The mutation responsible for this syndrome is expansion of the trinucleotide repeat-cytosine (C), adenine (A), guanine (G)-in a 5' translated portion of the androgen receptor (AR) gene from a normal, polymorphic length of n = 11-31 to n >/= 40. The resulting androgen receptor (AR) protein has an expanded polyglutamine tract in its NH(2)-terminal modulatory domain, and is postulated to lose a basic, intrinsic function that causes a mild form of androgen insensitivity; however, almost certainly, it also gains a novel, extrinsic function that is selectively neuronotoxic. The unexplained mechanism that culminates in this form of neuronspecific death is the prototype for three different adult-onset neuronopathies that are caused by (CAG)(n) expansions in other genes. PMID- 18407239 TI - Molecular characterization of nephrogenic diabetes insipidus. AB - Nephrogenic diabetes insipidus (NDI) is characterized by insensitivity of the distal renal nephron to the antidiuretic effect of the neurohypophyseal hormone arginine vasopressin. In the last 2 years, two different genetic defects causing the NDI phenotype have been identified. The genes involved encode proteins that reside at both ends of the cellular vasopressin signaling cascade, namely the vasopressin V(2) receptor and the aquaporin-2 water channel. Analysis of naturally occurring mutations in the V(2) receptor and the aquaporin-2 water channel will facilitate the study of structure-function correlates of both proteins, which will lead to substantial progress in elucidating the cellular mechanisms involved in the antidiuretic effect of vasopressin. PMID- 18407240 TI - Ecto-phosphatase activity on the external surface of Rhodnius prolixus salivary glands: modulation by carbohydrates and Trypanosoma rangeli. AB - The salivary glands of insect's vectors are target organs to study the vectors pathogens interactions. Rhodnius prolixus an important vector of Trypanosoma cruzi can also transmit Trypanosoma rangeli by bite. In the present study we have investigated ecto-phosphatase activity on the surface of R. prolixus salivary glands. Ecto-phosphatases are able to hydrolyze phosphorylated substrates in the extracellular medium. We characterized these ecto-enzyme activities on the salivary glands external surface and employed it to investigate R. prolixus-T. rangeli interaction. Salivary glands present a low level of hydrolytic activity (4.30+/-0.35 nmol p-nitrophenol (p-NP)xh(-1)xgland pair(-1)). The salivary glands ecto-phosphatase activity was not affected by pH variation; and it was insensitive to alkaline inhibitor levamisole and inhibited approximately 50% by inorganic phosphate (Pi). MgCl2, CaCl2 and SrCl2 enhanced significantly the ecto phosphatase activity detected on the surface of salivary glands. The ecto phosphatase from salivary glands surface efficiently releases phosphate groups from different phosphorylated amino acids, giving a higher rate of phosphate release when phospho-tyrosine is used as a substrate. This ecto-phosphatase activity was inhibited by carbohydrates as d-galactose and d-mannose. Living short epimastigotes of T. rangeli inhibited salivary glands ecto-phosphatase activity at 75%, while boiled parasites did not. Living long epimastigote forms induced a lower, but significant inhibitory effect on the salivary glands phosphatase activity. Interestingly, boiled long epimastigote forms did not loose the ability to modulate salivary glands phosphatase activity. Taken together, these data suggest a possible role for ecto-phosphatase on the R. prolixus salivary glands-T. rangeli interaction. PMID- 18407241 TI - Risk factors for complications after congenital cataract surgery without intraocular lens implantation in the first 18 months of life. AB - PURPOSE: To determine individual risk factors for the development of postoperative complications after pediatric cataract surgery in the first 18 months of life. DESIGN: Interventional, consecutive case series. METHODS: We retrospectively reviewed the records of 71 eyes of 46 children who underwent surgery for congenital cataract within the first 18 months of life. A limbal approach bimanual lens aspiration, posterior capsulorrhexis, and anterior vitrectomy without intraocular lens implantation was performed in all children. We examined the interrelationships of operative and postoperative complications with other variables such as patient age, family history, or ocular abnormalities. The mean follow-up period was 39 months. RESULTS: The most frequent postoperative complications were late-onset open-angle glaucoma (10.8%) and vitreous hemorrhage (10.8%), whereas early-onset glaucoma (4.6%) was less common. Secondary cataract was observed in seven eyes (9.2%). We determined a family history of aphakic glaucoma in first-degree relatives (P = .007) as well as cataract surgery in the first three months of life (P = .039) and nuclear cataracts (P = .0009) to be strong predictors of late-onset glaucoma. Secondary cataract formation was associated strongly with lensectomy in the first five months of life. The diagnosis of postoperative hemorrhages was associated significantly with the presence of persistent fetal vasculature (P < .0001). CONCLUSIONS: Patients with preoperative predictors at presentation such as young age at the time of surgery, a family history of aphakic glaucoma, nuclear cataract, or persistent fetal vasculature syndrome offer a clear target for extensive postoperative care after congenital cataract surgery. PMID- 18407242 TI - Endocytosis of sex steroids: the hypothesis of free hormones revisited. PMID- 18407243 TI - Neuroendocrinology: past, present and future. PMID- 18407244 TI - Macroergonomics as an organizing process for systems safety. AB - Hendrick is attributed with the formalization of organizational design and management (ODAM) in ergonomics [Hendrick, H.W., Kleiner, B.M., 2001. Macroergonomics: An Introduction to Work System Design. Human Factors and Ergonomics Society, Santa Monica, CA.]. Specifically, the method called "Macroergonomic Analysis of Structure" or MAS provides a framework and analysis of these factors and provides the context for an analysis of organizational design and management process through the MacroErgonomic Analysis and Design method (MEAD). Together, MAS and MEAD represent the formalization of staple methods in macroergonomics and can be used to organize existing tools and methods such as those that exist in systems safety and help to differentiate macroergonomics from other approaches. This article illustrates such an integrative role for macroergonomics with respect to systems safety using the example of the construction sector, a domain in which accidents, injuries and fatalities are all too common. PMID- 18407245 TI - Reducing cognitive vulnerability to depression: a preliminary investigation of MEmory Specificity Training (MEST) in inpatients with depressive symptomatology. AB - Reduced specificity of autobiographical memory retrieval represents an enduring vulnerability factor for depression. The present study is a preliminary evaluation of a newly developed group-based intervention program to increase specificity of memory retrieval. The MEmory Specificity Training (MEST) was administered on a weekly basis for 4 consecutive weeks to 10 inpatients with depressive symptomatology. Whereas earlier studies found that memory specificity does not improve following treatment as usual, the present results showed that participants' retrieval style became significantly more specific following MEST. These results suggest that the MEST may offer a potential and promising intervention to tackle a core cognitive process involved in depression and depressive vulnerability. PMID- 18407246 TI - Sulforaphane protects kidneys against ischemia-reperfusion injury through induction of the Nrf2-dependent phase 2 enzyme. AB - Reactive oxygen species are important mediators that exert a toxic effect during ischemia-reperfusion injury of various organs. Sulforaphane, which is a naturally occurring isothiocyanate that is present in cruciferous vegetables such as broccoli, is known to be an indirect antioxidant that acts by inducing Nrf2 dependent phase 2 enzymes. Phase 2 enzymes such as heme oxygenase-1, NAD(P)H: quinone oxidoreductase 1, glutathione reductase, and glutathione peroxidase participate in adaptive and protective responses to oxidative stress and various inflammatory stimuli. Therefore, we evaluated the preactivation of Nrf2 by sulforaphane to determine if it could inhibit ischemia-reperfusion-induced kidney damage. Treatment of HK2 renal tubular epithelial cells with sulforaphane effectively protected cells against cytotoxicity induced by hypoxia reoxygenation, and sulforaphane dramatically induced phase 2 enzymes by decreasing the Keap1 protein levels and increasing Nrf2 nuclear translocation. Additionally, a second set of experiments using a renal ischemia-reperfusion model produced results that were essentially the same as those observed when HK2 cells were used; namely, that sulforaphane induced Nrf2-dependent phase 2 enzymes and thereby improved ischemia-reperfusion-induced changes in the lipid hydroperoxides, glutathione, creatinine clearance, kidney weight, and histologic abnormalities. Collectively, these results suggest that sulforaphane can be used as an effective adjunct for the prevention of renal oxidative insults during ischemia-reperfusion injury. PMID- 18407247 TI - Short-term regulation of the Cl-/HCO3(-) exchanger in immortalized SHR proximal tubular epithelial cells. AB - The present study evaluated the activity of Cl(-)/HCO(3)(-) exchanger and the abundance of Slc26a6 in immortalized renal proximal tubular epithelial (PTE) cells from the Wistar-Kyoto rat (WKY) and spontaneously hypertensive rat (SHR) and identified the signaling pathways that regulate the activity of the transporter. The affinity for HCO(3)(-) was identical in WKY and SHR PTE cells, but V(max) values (in pH units/min) in SHR PTE cells (0.4016) were significantly higher than in WKY PTE cells (0.2304). The expression of Slc26a6 in SHR PTE cells was sevenfold that in WKY PTE cells. Dibutyryl-cAMP (db-cAMP) or forskolin, which increased endogenous cAMP, phorbol-12,13-dibutyrate (PDBu) and anisomycin, significantly (P<0.05) increased the Cl(-)/HCO(3)(-) exchanger activity in WKY and SHR PTE cells to a similar extent. The stimulatory effects of db-cAMP and forskolin were prevented by the PKA inhibitor H89, but not by chelerythrine. The stimulatory effects of PDBu were prevented by both chelerythrine and SB 203580, but not by H89 or the MEK inhibitor PD 98059. The stimulatory effect of anisomycin was prevented by SB 203580, but not by chelerythrine. Increases in phospho-p38 MAPK by anisomycin were identical in WKY and SHR PTE cells, this being sensitive to SB 203580 but not to chelerythrine. It is concluded that SHR PTE cells, which overexpress the Slc26a6 protein, are endowed with an enhanced activity of the Cl(-)/HCO(3)(-) exchanger. The Cl(-)/HCO(3)(-) exchanger is an effector protein for PKA, PKC and p38 MAPK in both WKY and SHR PTE cells. PMID- 18407249 TI - Visual-auditory spatial processing in auditory cortical neurons. AB - Neurons responsive to visual stimulation have now been described in the auditory cortex of various species, but their functions are largely unknown. Here we investigate the auditory and visual spatial sensitivity of neurons recorded in 5 different primary and non-primary auditory cortical areas of the ferret. We quantified the spatial tuning of neurons by measuring the responses to stimuli presented across a range of azimuthal positions and calculating the mutual information (MI) between the neural responses and the location of the stimuli that elicited them. MI estimates of spatial tuning were calculated for unisensory visual, unisensory auditory and for spatially and temporally coincident auditory visual stimulation. The majority of visually responsive units conveyed significant information about light-source location, whereas, over a corresponding region of space, acoustically responsive units generally transmitted less information about sound-source location. Spatial sensitivity for visual, auditory and bisensory stimulation was highest in the anterior dorsal field, the auditory area previously shown to be innervated by a region of extrastriate visual cortex thought to be concerned primarily with spatial processing, whereas the posterior pseudosylvian field and posterior suprasylvian field, whose principal visual input arises from cortical areas that appear to be part of the 'what' processing stream, conveyed less information about stimulus location. In some neurons, pairing visual and auditory stimuli led to an increase in the spatial information available relative to the most effective unisensory stimulus, whereas, in a smaller subpopulation, combined stimulation decreased the spatial MI. These data suggest that visual inputs to auditory cortex can enhance spatial processing in the presence of multisensory cues and could therefore potentially underlie visual influences on auditory localization. PMID- 18407248 TI - Expression of GABA A receptor alpha1 subunit mRNA and protein in rat neocortex following photothrombotic infarction. AB - Photothrombotic infarcts of the neocortex result in structural and functional alterations of cortical networks, including decreased GABAergic inhibition, and can generate epileptic seizures within 1 month of lesioning. In our study, we assessed the involvement and potential changes of cortical GABA A receptor (GABA AR) alpha1 subunits at 1, 3, 7, and 30 days after photothrombosis. Quantitative competitive reverse transcription-polymerase chain reaction (cRT-PCR) and semi quantitative Western blot analysis were used to investigate GABA AR alpha1 subunit mRNA and protein levels in proximal and distal regions of perilesional cortex and in homotopic areas of young adult Sprague-Dawley rats. GABA AR alpha1 subunit mRNA levels were decreased ipsilateral and contralateral to the infarct at 7 days, but were increased bilaterally at 30 days. GABA AR alpha1 subunit protein levels revealed no significant change in neocortical areas of both hemispheres of lesioned animals compared with protein levels of sham-operated controls at 1, 3, 7, and 30 days. At 30 days, GABA AR alpha1 subunit protein expression was significantly increased in lesioned animals within proximal and distal regions of perilesional cortex compared with distal neocortical areas contralaterally (Student's t-test, p<0.05). Short- and long-term alterations of mRNA and protein levels of the GABA AR alpha1 subunit ipsilateral and contralateral to the lesion may influence alterations in cell surface receptor subtype expression and GABA AR function following ischemic infarction and may be associated with formative mechanisms of poststroke epileptogenesis. PMID- 18407250 TI - Distribution of estrogen receptor 2 mRNAs (Esr2a and Esr2b) in the brain and pituitary of the sea bass (Dicentrarchus labrax). AB - Three different estrogen receptors (ERs) have been characterized in teleost fish, i.e. Esr1, Esr2a and Esr2b. In this study we carried out in situ hybridizations in the brain and pituitary of the sea bass (Dicentrarchus labrax) to study the putative involvement of Esr2 subtypes in the control of gonadotropin secretion in fish. Our studies demonstrated that both receptors are expressed within the main hypophysiotrophic areas of the sea bass brain thus providing neuroanatomical basis for the involvement of Esr2 subtypes in the long (or indirect) regulatory feedbacks on pituitary function in the sea bass. The results revealed that Esr2b mRNA distribution was restricted to the preoptic area and tuberal hypothalamus. On the contrary, Esr2a presented a widespread distribution and transcripts were detected within the ventral telencephalon, preoptic area, hypothalamus, thalamus, posterior tubercle, mesencephalon and rhombencephalon. New Esr2-expressing areas were described in all of the above areas. This paper is the first demonstration of Esr2a and Esr2b expression in the follicle-stimulating hormone beta-subunit (betaFSH)- and luteinizing hormone beta-subunit (betaLH)-expressing cells in the fish pituitary, thus suggesting the participation of both receptors in the direct effect of estrogen on the control of gonadotropin hormone synthesis. PMID- 18407251 TI - GABA homeostasis contributes to the developmental programming of anxiety-related behavior. AB - During development, when inhibitory and excitatory synapses are formed and refined, homeostatic mechanisms act to adjust inhibitory input in order to maintain neural activity within a normal range. As the brain matures, synaptogenesis slows and a relatively stable level of inhibition is achieved. Deficits in inhibitory neurotransmission are associated with increased anxiety related behavior and drugs that potentiate GABA function, the major inhibitory neurotransmitter in the brain, are effective anxiolytics. These observations raise the possibility that transient perturbations in the activity of neural circuits during development might induce compensatory changes in inhibition that could persist into adulthood and contribute to changes in anxiety-related behavior. To test this hypothesis, we treated mice continuously during the major period of forebrain synaptogenesis (P14-28) with the GABA-A receptor positive modulator diazepam and assessed anxiety-related behavior in adulthood. Control experiments confirmed anxiolytic effects of the drug following one day of treatment and the development of tolerance following two weeks of treatment. When tested in adulthood, one month after the end of treatment, diazepam-treated mice exhibited significantly increased behavioral inhibition in the open-field, elevated-plus maze, and novel object behavioral paradigms. Levels of benzodiazepine binding sites in amygdala and frontal cortex were specifically decreased in diazepam-treated mice demonstrating that homeostatic adjustments in GABA function persist into adulthood. Our results show that increased GABAergic activity can affect the developmental programming of anxiety-related behavior. PMID- 18407252 TI - NMDA receptors in the basolateral amygdala and gustatory neophobia. AB - The attenuation of gustatory neophobia occurs during repeated exposures to an initially novel taste solution that is increasingly perceived as safe and familiar. The present study examined whether NMDA receptors in the basolateral region of the amygdala (BLA) are involved in this important behavioral phenomenon. The results, which show that the attenuation, but not initial occurrence, of gustatory neophobia is dependent upon NMDA receptors in the BLA, are discussed with reference to a similar finding involving NMDA receptors in the insular cortex. PMID- 18407253 TI - Establishment of an immortalized GABAergic neuronal progenitor cell line from embryonic ventral mesencephalon in the rat. AB - Effective cell replacement therapies for neurological disease require neuron restricted precursors as grafted cells. The problem of obtaining sufficient grafts for transplantation can be resolved by creating an appropriate immortalized cell line. In the present study, a thermally controlled immortalized GABAergic neuronal progenitor cell line (RMNE6) was established from E13 rat ventral mesencephalon cells immortalized using the temperature-sensitive mutant of SV40 large T antigen (ts-TAg). RMNE6 cells proliferated rapidly and expressed a neuron-like phenotype at the permissive temperature (33 degrees C), but eventually stopped growing at the non-permissive temperature (39 degrees C). Expression of the neuronal markers PSA-NCAM, beta-tubulin III and MAP2 by RMNE6 cells was confirmed by RT-PCR or immunocytochemistry. Furthermore, these cells exhibited functional GABAergic neuron properties, as evidenced by the expression of glutamate decarboxylase (GAD) as well as the synthesis and release of the neurotransmitter GABA in a calcium-dependent manner. Moreover, RMNE6 cells spontaneously expressed and secreted several neurotrophic factors, such as NGF, BDNF, NT-3, NT-4/5, and GDNF. The cells survived well and kept expression of SV40 Tag, GAD65/67 and GABA in the striatum, at least 28 days after being transplanted in the rat brain. Tumorigenesis assays confirmed the safety of the immortalized cell line in vivo. Taken together, the results support the use of RMNE6 cells as an ideal cell model for transplantation research aimed at the treatment and prevention of neurodegenerative disease. PMID- 18407254 TI - GABAergic and non-GABAergic thalamic, hypothalamic and basal forebrain projections to the ventral oral pontine reticular nucleus: their implication in REM sleep modulation. AB - The ventral part of the oral pontine reticular nucleus (vRPO) is a demonstrated site of brainstem REM-sleep generation and maintenance. The vRPO has reciprocal connections with structures that control other states of the sleep-wakefulness cycle, many situated in the basal forebrain and the diencephalon. Some of these connections utilize the inhibitory neurotransmitter GABA. The aim of the present work is to map the local origin of the basal forebrain and diencephalon projections to the vRPO whether GABAergic or non-GABAergic. A double-labelling technique combining vRPO injections of the neuronal tracer, cholera-toxin (CTB), with GAD-immunohistochemistry, was used for this purpose in adult cats. All of the numerous CTB-positive neurons in the reticular thalamic and dorsocaudal hypothalamic nuclei were double-labelled (CTB/GAD-positive) neurons. Approximately 15%, 14% and 16% of the CTB-positive neurons in the zona incerta and the dorsal and lateral hypothalamic areas are, respectively, CTB/GAD-positive neurons. However, only some double-labelled neurons were found in other hypothalamic nuclei with abundant CTB-positive neurons, such as the paraventricular nucleus, perifornical area and H1 Forel field. In addition, CTB positive neurons were abundant in the central amygdaline nucleus, terminal stria bed nuclei, median preoptic nucleus, medial and lateral preoptic areas, dorsomedial and ventromedial hypothalamic nuclei, posterior hypothalamic area and periventricular thalamic nucleus. The GABAergic and non-GABAergic connections described here may be the morphological pillar through which these prosencephalic structures modulate, either by inhibiting or by exciting, the vRPO REM-sleep inducing neurons during the different sleep-wakefulness cycle states. PMID- 18407255 TI - Hyperbaric oxygen preconditioning induces tolerance against brain ischemia reperfusion injury by upregulation of antioxidant enzymes in rats. AB - The present study examined the hypothesis that cerebral ischemic tolerance induced by hyperbaric oxygen preconditioning (HBO-PC) is associated with an increase of antioxidant enzyme activity. Male Sprague-Dawley rats (250-280 g, n=74) were divided into sham, middle cerebral artery occlusion (MCAO) for 90 min, and MCAO plus HBO-PC groups. HBO-PC was conducted four times by given 100% oxygen at 2.5 atmosphere absolute (ATA), for 1 h at every 12 h interval for 2 days. At 24 h after the last HBO-PC, MCAO was performed and at 24 h after MCAO, neurological function and Nissl Staining were performed to evaluate the effect of HBO-PC. Malondialdehyde (MDA) content, activity of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) sampled from the hippocampus, ischemic penumbra or core of cortex were measured. HBO-PC decreased mortality rate, improved neurological recovery, lessened neuronal injury, reduced the level of MDA and increased the antioxidant activity of CAT and SOD. These observations demonstrated that an upregulation of the antioxidant enzyme activity by HBO preconditioning plays an important role in the generation of tolerance against brain ischemia-reperfusion injury. PMID- 18407256 TI - Daughterless dictates Twist activity in a context-dependent manner during somatic myogenesis. AB - Somatic myogenesis in Drosophila relies on the reiterative activity of the basic helix-loop-helix transcriptional regulator, Twist (Twi). How Twi directs multiple cell fate decisions over the course of mesoderm and muscle development is unclear. Previous work has shown that Twi is regulated by its dimerization partner: Twi homodimers activate genes necessary for somatic myogenesis, whereas Twi/Daughterless (Da) heterodimers lead to the repression of these genes. Here, we examine the nature of Twi/Da heterodimer repressive activity. Analysis of the Da protein structure revealed a Da repression (REP) domain, which is required for Twi/Da-mediated repression of myogenic genes, such as Dmef2, both in tissue culture and in vivo. This domain is crucial for the allocation of mesodermal cells to distinct fates, such as heart, gut and body wall muscle. By contrast, the REP domain is not required in vivo during later stages of myogenesis, even though Twi activity is required for muscles to achieve their final pattern and morphology. Taken together, we present evidence that the repressive activity of the Twi/Da dimer is dependent on the Da REP domain and that the activity of the REP domain is sensitive to tissue context and developmental timing. PMID- 18407257 TI - Ndrg4 is required for normal myocyte proliferation during early cardiac development in zebrafish. AB - NDRG4 is a novel member of the NDRG family (N-myc downstream-regulated gene). The roles of NDRG4 in development have not previously been evaluated. We show that, during zebrafish embryonic development, ndrg4 is expressed exclusively in the embryonic heart, the central nervous system (CNS) and the sensory system. Ndrg4 knockdown in zebrafish embryos causes a marked reduction in proliferative myocytes and results in hypoplastic hearts. This growth defect is associated with cardiac phenotypes in morphogenesis and function, including abnormal heart looping, inefficient circulation and weak contractility. We reveal that ndrg4 is required for restricting the expression of versican and bmp4 to the developing atrioventricular canal. This constellation of ndrg4 cardiac defects phenocopies those seen in mutant hearts of heartstrings (hst), the tbx5 loss-of-function mutants in zebrafish. We further show that ndrg4 expression is significantly decreased in hearts with reduced tbx5 activities. Conversely, increased expression of tbx5 that is due to tbx20 knockdown leads to an increase in ndrg4 expression. Together, our studies reveal an essential role of ndrg4 in regulating proliferation and growth of cardiomyocytes, suggesting that ndrg4 may function downstream of tbx5 during heart development and growth. PMID- 18407258 TI - Divergent functions of orthodenticle, empty spiracles and buttonhead in early head patterning of the beetle Tribolium castaneum (Coleoptera). AB - The head gap genes orthodenticle (otd), empty spiracles (ems) and buttonhead (btd) are required for metamerization and segment specification in Drosophila. We asked whether the function of their orthologs is conserved in the red flour beetle Tribolium castaneum which in contrast to Drosophila develops its larval head in a way typical for insects. We find that depending on dsRNA injection time, two functions of Tc-orthodenticle1 (Tc-otd1) can be identified. The early regionalization function affects all segments formed during the blastoderm stage while the later head patterning function is similar to Drosophila. In contrast, both expression and function of Tc-empty spiracles (Tc-ems) are restricted to the posterior part of the ocular and the anterior part of the antennal segment and Tc buttonhead (Tc-btd) is not required for head cuticle formation at all. We conclude that the gap gene like roles of ems and btd are not conserved while at least the head patterning function of otd appears to be similar in fly and beetle. Hence, the ancestral mode of insect head segmentation remains to be discovered. With this work, we establish Tribolium as a model system for arthropod head development that does not suffer from the Drosophila specific problems like head involution and strongly reduced head structures. PMID- 18407260 TI - Hoxd13 binds in vivo and regulates the expression of genes acting in key pathways for early limb and skeletal patterning. AB - 5' HoxD genes are required for the correct formation of limb skeletal elements. Hoxd13, the most 5'-located HoxD gene, is important for patterning the most distal limb region, and its mutation causes human limb malformation syndromes. The mechanisms underlying the control of developmental processes by Hoxd13, and by Hox genes in general, are still elusive, due to the limited knowledge on their direct downstream target genes. We identified by ChIP-on-chip 248 known gene loci bound invivo by Hoxd13. Genes relevant to limb patterning and skeletogenesis were further analysed. We found that Hoxd13 binds invivo, in developing limbs, the loci of Hand2, a gene crucial to limb AP axis patterning, of Meis1 and Meis2, involved in PD patterning, of the Sfrp1, Barx1, and Fbn1 genes, involved in skeletogenesis, and of the Dach1, Bmp2, Bmp4, andEmx2 genes. We show that Hoxd13 misexpression in developing chick limbs alters the expression of the majority of these genes, supporting the conclusion that Hoxd13 directly regulates their transcription. Our results indicate that 5' Hox proteins regulate directly both key genes for early limb AP and PD axis patterning and genes involved, at later stages, in skeletal patterning. PMID- 18407259 TI - Septate junctions are required for ommatidial integrity and blood-eye barrier function in Drosophila. AB - The anatomical organization of the Drosophila ommatidia is achieved by specification and contextual placement of photoreceptors, cone and pigment cells. The photoreceptors must be sealed from high ionic concentrations of the hemolymph by a barrier to allow phototransduction. In vertebrates, a blood-retinal barrier (BRB) is established by tight junctions (TJs) present in the retinal pigment epithelium and endothelial membrane of the retinal vessels. In Drosophila ommatidia, the junctional organization and barrier formation is poorly understood. Here we report that septate junctions (SJs), the vertebrate analogs of TJs, are present in the adult ommatidia and are formed between and among the cone and pigment cells. We show that the localization of Neurexin IV (Nrx IV), a SJ-specific protein, coincides with the location of SJs in the cone and pigment cells. Somatic mosaic analysis of nrx IV null mutants shows that loss of Nrx IV leads to defects in ommatidial morphology and integrity. nrx IV hypomorphic allelic combinations generated viable adults with defective SJs and displayed a compromised blood-eye barrier (BEB) function. These findings establish that SJs are essential for ommatidial integrity and in creating a BEB around the ion and light sensitive photoreceptors. Our studies may provide clues towards understanding the vertebrate BEB formation and function. PMID- 18407261 TI - Catfish consumption as a contributor to elevated PCB levels in a non-Hispanic black subpopulation. AB - The human body burden of polychlorinated biphenyls (PCBs) sharply declined after production was banned in the US in 1979. For the 10% of the US population that remains most exposed to PCBs, fish consumption is the primary source. National Health and Nutrition Examination Survey (NHANES) data indicates that the highest remaining PCB levels exist in a non-Hispanic black subpopulation. Our review suggests that catfish consumption may be a significant PCB source for the one million non-Hispanic black anglers who fish for catfish. In comparison to non Hispanic white anglers, non-Hispanic black anglers consume more catfish, are more likely to eat the whole fish rather than just the fillets that contain less PCBs, and are more likely to fish in watersheds with high PCB contamination. Efforts to diminish potential racial disparities in PCB exposure are challenged by geographic, economic, cultural, and educational barriers. In response, we propose that a fish consumption survey be performed that identifies the extent of subsistence fishing by non-Hispanic black anglers for catfish in watersheds with PCB contamination, the type and quantity of catfish subsistence fishing provides, and what actions would help moderate PCB exposure due to subsistence fishing for catfish in such areas. Understanding the contamination and consumption factors that contribute to higher PCB body burdens will help identify and offer solutions to racial disparities in exposure to PCBs due to subsistence fishing while providing a model to prevent similar disparities in exposure to toxics ranging from mercury to polybrominated diphenyl ethers. PMID- 18407262 TI - Brain insulin, energy and glucose homeostasis; genes, environment and metabolic pathologies. AB - The central nervous system is essential in maintaining energy and glucose homeostasis. In both animals and humans, efficient cerebral insulin signalling is a pivotal control element in these pathophysiological processes. The action of insulin in the brain is under a multilevel control via metabolic, endocrine and neural signals induced by nutrients, integrated mainly by the hypothalamus. Of particular interest is the interaction of insulin with the anabolic and catabolic neuroregulators. The anorexic peptides insulin, leptin and the neurotransmitter serotonin share common signalling pathways involved in food intake, in particular the insulin receptor substrate, phosphatidylinositol-3-kinase (PI3K) pathway. The dialogue of neurotransmitters and peptides via this signalling pathway is potentially of major importance in the pathophysiology of the brain in general and specifically in the regulation of feeding behaviour. At this time, a new concept in the aetiopathology of type 2 diabetes is immerging. This concept proposes that the combination of defective pancreatic beta-cell function and insulin resistance not only in classical insulin target tissues but in every tissue, contributes to the onset of the disease. It highlights the importance of the disruption of cerebral insulin signal transmission and its direct relation to metabolic diseases. Impaired brain insulin signalling, a link coupling obesity to diabetes, may be related to either genetic factors, or environmental factors such as stress, over or under-feeding and unbalanced diets: such factors may work either independently or in concert. Current approaches used for the prevention and treatment of type 2 diabetes are not adequately effective. Most of the anti diabetic therapies induce many adverse effects, in particular obesity, and thus may initiate a vicious cycle of problems. In order to develop new, more efficient, preventive and therapeutic strategies for metabolic pathologies, there is an urgent need for increased understanding of the complexity of insulin signalling in the brain and on the interactive, central and peripheral effects of insulin. PMID- 18407263 TI - Erythropoietin in cardiac disease: new features of an old drug. AB - Erythropoietin is a haematopoietic hormone with extensive non-haematopoietic effects. The discovery of an erythropoietin receptor outside the haematopoietic system has fuelled the research into the beneficial effects of erythropoietin for various conditions, predominantly in cardiovascular disease. Experimental evidence has revealed the cytoprotective and angiogenic properties of erythropoietin and it seems that the erythropoietin-erythropoietin receptor system provides a powerful backbone against acute and chronic myocardial ischemia, each gaining from the different properties of erythropoietin. Clinical trials in which erythropoietin was titrated to achieve certain haematocrit levels have generated equivocal results. It has been suggested that a (too) high haematocrit is undesirable in cardiovascular disease. We have shown that intermittent (low-dose) erythropoietin administration, that does not increase haematocrit substantially, suffices to activate the beneficial downstream pathways of erythropoietin. We postulate that intermittent administration or a lower than conventional dose of erythropoietin, not only aimed at increasing haemoglobin at high levels, will provide powerful cellular protection and will improve cardiac outcome, without the side effects of erythropoietin associated with increased haematocrit. PMID- 18407264 TI - Structural brain imaging in diabetes: a methodological perspective. AB - Brain imaging provides information on brain anatomy and function and progression of cerebral abnormalities can be monitored. This may provide insight into the aetiology of diabetes related cerebral disorders. This paper focuses on the methods for the assessment of white matter hyperintensities and brain atrophy on structural brain images, mostly magnetic resonance imaging, in diabetes. These methods range from visual rating scales to advanced semi-automated and automated image processing techniques such as volumetry and voxel-based morphometry. The findings of previous imaging studies in diabetes are discussed from a methodological perspective and recommendations for future research are given. PMID- 18407265 TI - Composition and function of the Crumbs protein complex in the mammalian retina. AB - The Crumbs proteins (CRBs) are transmembrane proteins, homologous to Drosophila Crumbs, with a key role in defining the apical membrane domain in photoreceptors as well as in embryonic epithelia. Crumbs proteins are conserved between species and their intracellular domains are involved in organizing a conserved macromolecular protein scaffold with important roles in cell polarity as well as morphogenesis and maintenance of the retina. Mutations in the gene encoding human CRB1, the first one identified out of the three human orthologs, have been associated with a number of retinal dystrophies including Leber amaurosis and retinitis pigmentosa type 12. Although no other mammalian Crumbs complex members as of yet have been associated with retinal degeneration, disruption of different zebrafish and fruitfly orthologs can lead to various retinal defects. The core Crumbs complex localizes apical to the outer limiting membrane, where photoreceptors and Muller glia contact each other. Correct functioning of Crumbs ensures adhesion between these cells by an unknown mechanism. This review summarizes the current view on the composition and function of the Crumbs prsotein complex in the mammalian retina. Recently, a number of new members of the Crumbs protein complex have been identified. These include most members of the membrane palmitoylated protein family (MPP), involved in assembly of macromolecular protein complexes. Some components of the complex are found to exert a function in the photoreceptor synapses and/or at the region of the connecting cilium. Studies using polarized cell cultures or model organisms, like Drosophila and zebrafish, suggest important links of the Crumbs protein complex to several biological processes in the mammalian eye, including retinal patterning, ciliogenesis and vesicular transport. PMID- 18407266 TI - Molecular identification and localization of P2X receptors in the rat lens. AB - There is growing evidence suggesting that purine-mediated signaling pathways play important roles in the ocular lens. We have previously reported the expression of P2Y receptors in the lens; and in this present study, we show that P2X receptors are also expressed. Transcripts for P2X1-7 receptors were detected in lens fiber cells using the reverse-transcription polymerase chain reaction (RT-PCR). Western blot analysis confirmed the expression of all P2X receptor subtypes at the protein level. Immunohistochemistry revealed P2X1 and P2X7 expression in the cytoplasm of cortical fiber cells. P2X2 expression was confined to the apical apical interface between epithelial and fiber cells with little expression in cortical fiber cells. In contrast, P2X3, P2X4, P2X5 and P2X6 were expressed throughout the lens extending from the outer cortex through to the core. In the outer cortex, immunolocalisation of these P2X receptors was predominantly cytoplasmic. However, deeper into the lens, P2X receptor immunolabeling became more membranous, indicating the recruitment of P2X receptors from cytoplasmic vesicles into the membranes of mature fiber cells in the lens core. Western blotting confirmed regional differences in P2X receptor expression. The differential expression of P2X receptors in the ocular lens indicates that the purinergic signaling pathways may be involved in the maintenance of lens homeostasis. PMID- 18407267 TI - Infectious implications in the porcine model of natural orifice transluminal endoscopic surgery (NOTES) with PEG-tube closure: a quantitative bacteriologic study. AB - BACKGROUND: Obtaining reliable closure of transvisceral defects currently limits natural orifice transluminal endoscopic surgery (NOTES). PEG tubes are potential means of managing NOTES gastrotomies. OBJECTIVE: To determine the efficacy of PEG closure after NOTES. DESIGN: An experimental animal study. SETTING: A laboratory. INTERVENTIONS: The pigs received gastric lavage with saline solution, chloramphenicol, or no lavage, and then underwent transgastric NOTES peritoneoscopy. Cultures were obtained by endoscopy during the surgery. A sterile foreign body was left in the peritoneal cavity. The gastrotomy was closed with a 20F PEG tube. The animals were observed for 14 days and underwent sterile laparotomy. Cultures of the foreign body and the peritoneal cavity were obtained. MAIN OUTCOME MEASUREMENTS: Abscess rate, peritoneal quantitative cultures. RESULTS: After 1 exclusion for anesthetic complications, 19 animals underwent NOTES; 18 (94.7%) survived the entire postoperative period. One animal died on postoperative day 2 after the PEG tube dislodged. At 14 days, 5 animals (27.8%) had intra-abdominal abscesses, 8 (44.4%) had positive peritoneal cultures, and 9 (50%) foreign bodies were contaminated on culture. Infectious complications were not altered by the type of gastric lavage or peritoneal bacterial inoculum introduced at the time of surgery. LIMITATION: An animal model. CONCLUSIONS: PEG closure of a NOTES gastrotomy is associated with subclinical intra-abdominal abscess formation and can result in death when the tube is dislodged during the early postoperative period. Preprocedural gastric lavage does not alter the intra abdominal bacterial burden introduced at the time of surgery or subsequent infectious outcomes in the porcine model. These concerning findings necessitate additional studies to determine if porcine models are appropriate and applicable to human subjects in the NOTES setting. PMID- 18407268 TI - Mask adaptor--a novel method of positive pressure ventilation during propofol deep sedation for upper GI endoscopy. AB - BACKGROUND: Propofol dosages required for upper GI endoscopy are often high enough to pose serious risks of respiratory depression. Stopping the procedure and bag ventilating a patient until the propofol wears off may be a safer management because traditional mask ventilation is not available. OBJECTIVE: We introduce the mask adaptor for upper GI endoscopy (MAUGE), a new method of positive pressure ventilation during upper GI endoscopy, and assessed its feasibility and safety. DESIGN: Subjects received propofol 1.5 to 2.5 mg/kg injection followed by repeated doses of 20 to 30 mg if necessary. SETTING: Tertiary hospital. PATIENTS: Thirty patients, American Society of Anesthesiologists class I to III, undergoing upper GI endoscopy and requesting sedation. INTERVENTIONS: After connecting the MAUGE to the anesthetic ventilation circuit and mask, the endoscope was inserted into the patient's digestive tract through the channel for endoscopes in the MAUGE and through the mask. Oxygen was supplied to the respiratory tract through the channel for gas in the MAUGE and through the mask by using positive pressure ventilation by bag-valve-mask ventilation. MAIN OUTCOME MEASUREMENTS: Heart rate, noninvasive blood pressure, end-tidal carbon dioxide tension, oxygen saturation, respiratory waveform. RESULTS: Oxygen saturation was more than 95% throughout the endoscopy in all patients. Positive ventilation was achieved in all patients and consistent with thoracic wall movement and respiratory waveforms shown by capnography. LIMITATIONS: The MAUGE cannot seal the respiratory tract. Patients in high risk for aspiration should not be considered candidates for using the MAUGE. CONCLUSIONS: By use of the MAUGE, positive pressure ventilation was efficaciously achieved, and desaturation and carbon dioxide retention were effectively avoided during the upper GI endoscopy procedure. PMID- 18407269 TI - Endoscopic removal of bile-duct stones by using a rotatable papillotome and a large-balloon dilator in patients with a Billroth II gastrectomy (with video). AB - BACKGROUND: Endoscopic stone removal is difficult in patients with a Billroth II gastrectomy. OBJECTIVE: To evaluate the usefulness of a rotatable papillotome and large-balloon dilation for removing bile-duct stones in patients with a Billroth II gastrectomy. DESIGN: A case series. SETTING: A large tertiary-referral center. PATIENTS AND INTERVENTION: Nine patients with bile-duct stones and a previous Billroth II gastrectomy were included. An endoscopic sphincterotomy (EST) was performed with a rotatable papillotome that could correct the axis of the cut toward the 5-o'clock direction; a large-balloon dilation (LBD) was then performed. MAIN OUTCOME MEASUREMENTS: The ability to perform an EST in the desired direction, successful stone removal, and complications. RESULTS: Rotation of the papillotome toward the 5-o'clock direction and an EST were achieved in 8 patients (89%). Stones were removed by EST and LBD in all 8 patients. There were no complications. LIMITATION: A small sample size. CONCLUSIONS: Limited EST by using a rotatable papillotome plus large-balloon dilation seemed to be safe, easy, and effective for removing bile-duct stones in patients with a Billroth II gastrectomy. PMID- 18407270 TI - Cost-effectiveness analysis of management strategies for obscure GI bleeding. AB - BACKGROUND AND AIMS: Of patients who are seen with GI hemorrhage, approximately 5% will have a small-bowel source. Management of these patients entails considerable expense. We performed a decision analysis to explore the optimal management strategy for obscure GI hemorrhage. METHODS: We used a cost effectiveness analysis to compare no therapy (reference arm) to 5 competing modalities for a 50-year-old patient with obscure overt bleeding: (1) push enteroscopy, (2) intraoperative enteroscopy, (3) angiography, (4) initial anterograde double-balloon enteroscopy (DBE) followed by retrograde DBE if the patient had ongoing bleeding, and (5) small-bowel capsule endoscopy (CE) followed by DBE guided by the CE findings. The model included prevalence rates for small bowel lesions, sensitivity for each intervention, and the probability of spontaneous bleeding cessation. We examined total costs and quality-adjusted life years (QALY) over a 1-year time period. RESULTS: An initial DBE was the most cost effective approach. The no-therapy arm cost $532 and was associated with 0.870 QALYs compared with $2407 and 0.956 QALYs for the DBE approach, which resulted in an incremental cost-effectiveness ratio of $20,833 per QALY gained. Compared to the DBE approach, an initial CE was more costly and less effective. The initial DBE arm resulted in an 86% bleeding cessation rate compared to 76% for the CE arm and 59% for the no-therapy arm. The model results were robust to a wide range of sensitivity analyses. LIMITATIONS: The short time horizon of the model, because of the lack of long-term data about the natural history of rebleeding from small intestinal lesions. CONCLUSIONS: An initial DBE is a cost-effective approach for patients with obscure bleeding. However, capsule-directed DBE may be associated with better long-term outcomes because of the potential for fewer complications and decreased utilization of endoscopic resources. PMID- 18407271 TI - Cost-effectiveness analysis of high-dose omeprazole infusion before endoscopy for patients with upper-GI bleeding. AB - BACKGROUND: The use of intravenous (i.v.) proton pump inhibitors (PPI) before an endoscopy in upper-GI bleeding (UGIB) was shown to reduce the need of endoscopic therapy and shorten hospital stay. OBJECTIVE: To investigate whether preemptive use of a PPI in UGIB is a cost-effective strategy. DESIGN: A decision analysis model that represents treatment pathways for patients with UGIB was constructed and structuralized by 30-day outcomes. Direct costs of medical treatment, diagnostic and therapeutic endoscopy, endoscopic re-treatment, surgery, and hospitalization were analyzed. SETTING: Prince of Wales Hospital, Hong Kong. PATIENTS: A total of 631 patients were recruited. Sixty patients (19.1%) in the PPI group and 90 patients (28.4%) in the placebo group required endoscopic hemostasis at index endoscopy. MAIN OUTCOME MEASUREMENTS: The primary measurements were cost-effectiveness ratios and incremental cost-effectiveness ratios (ICER) to avert endoscopic therapy between PPI and placebo treatment. Sensitivity analyses were conducted by varying the cost of endoscopy, hospitalization, the incidence rate of endoscopic therapy, and the proportion of bleeding peptic ulcers. RESULTS: The overall direct cost per patient was U.S. dollars (USD) $2813 for PPI treatment and USD $2948 for the placebo. A PPI reduced endoscopic therapy by 7.4% and resulted in a lower cost-effectiveness ratio per endoscopic therapy averted (USD $3561) than the placebo (USD $4117). The ICER value was USD -$1843, which indicated that preemptive PPI treatment is more effective and less costly for UGIB. When the proportions of patients with peptic ulcer bleeding were greater than 8.3%, the preemptive PPI treatment remained cost saving. CONCLUSIONS: Preemptive use of IV PPI before an endoscopy is a cost-effective strategy in the management of UGIB. PMID- 18407272 TI - Sex steroids and metabolic parameter levels in a seasonal breeding fish (Sparus aurata L.). AB - The gilthead seabream is a protandrous hermaphrodite seasonal breeding teleost with a bisexual gonad that offers an interesting model for studying the two first reproductive cycles (RCs), in which the specimens develops as males. During the first RC (RC1), 11-ketotestosterone (11-KT) and testosterone (T), the main androgens in fish, play different and specific roles in the testicular physiology as they peak at different stages of RC1. Moreover, the profiles of T serum levels during the second RC (RC2) demonstrated that this androgen is not essential in the testicular regression process that occurs during this cycle. However, changes in serum levels suggest that 17beta-estradiol (E2) orchestrates this process during both RCs. Moreover, the E2 serum levels recorded during RC2 indicate that there is a threshold level that determines the initiation of ovarian development during this cycle without promoting complete feminization. We analysed triglyceride, protein, glucose and lactate serum levels in order to establish a relationship between the mobilization and transfer of nutrients, the hormonal changes that take place during the RC and body composition, finding that in vivo serum levels of metabolites change significantly throughout the first two RCs, although the physiological relevance of such changes is still unknown. Triglyceride levels seem to be affected by the beginning of ovary development during RC2 but not by testicular recrudescence during RC1. Moreover, glucose and lactate might be important sources of energy during the resting and testicular involution stages. PMID- 18407274 TI - Mathematical models of the simplest fuzzy PI/PD controllers with skewed input and output fuzzy sets. AB - This paper unveils mathematical models for fuzzy PI/PD controllers which employ two skewed fuzzy sets for each of the two-input variables and three skewed fuzzy sets for the output variable. The basic constituents of these models are Gamma type and L-type membership functions for each input, trapezoidal/triangular membership functions for output, intersection/algebraic product triangular norm, maximum/drastic sum triangular conorm, Mamdani minimum/Larsen product/drastic product inference method, and center of sums defuzzification method. The existing simplest fuzzy PI/PD controller structures derived via symmetrical fuzzy sets become special cases of the mathematical models revealed in this paper. Finally, a numerical example along with its simulation results are included to demonstrate the effectiveness of the simplest fuzzy PI controllers. PMID- 18407273 TI - Androgens in health and disease: an overview. PMID- 18407275 TI - Temporary intravascular shunts used as a damage control surgery adjunct in complex vascular injury: collective review. AB - In this systemic review, we summarise the types, configurations, durations, indications and complications of the temporary intravascular shunts used as an adjunct of damage control surgery (DCS) in severe vascular injuries. We conclude that temporary intravascular shunts can be used without systemic anticoagulation for a prolonged time to maintain distal perfusion in combined orthopaedic and vascular injuries, in the setting of DCS and transferring. PMID- 18407277 TI - Fixation versus hemiarthroplasty for undisplaced intracapsular hip fractures. AB - The conventional treatment for an undisplaced intracapsular hip fracture is internal fixation. A possible alternative treatment is replacement hemiarthroplasty. We have compared the outcomes of an age, sex and co-morbidity matched cohort of 346 patients who have had their undisplaced intracapsular hip fracture treated using cannulated screws with a group of 346 patients who have had a displaced intracapsular fracture treated using a hemiarthroplasty. Patients treated by internal fixation had a shorter operation time (43 versus 67 min), reduced orthopaedic ward stay (11 versus 15 days), lower incidence of peri operative complications (24 versus 81), and a lower 1-year mortality (19% versus 26%). Additional benefits for the fixation group were less pain at 1 year, less reduction in mobility and lower dependence on walking aids. All these differences were statistically significant. Hemiarthroplasty had a lower re-admission rate (43 versus 14 cases) and re-operation rate (59 versus 22 cases). These results support the use of internal fixation for undisplaced intracapsular fractures and confirm that the final outcome for an undisplaced intracapsular hip fracture is significantly better than for a displaced intracapsular fracture, despite a higher re-operation rate after internal fixation. PMID- 18407276 TI - Troponin T as a predictive marker of morbidity in patients with fractured neck of femur. AB - INTRODUCTION: This study aims prospectively to assess perioperative measurement of Troponin T, a marker of myocardial injury, as a predictor of morbidity and mortality in patients undergoing surgery for fractured neck of femur. METHOD: All patients aged 65 years and over presenting with a fractured neck of femur over a 4-month period were initially included. Exclusion criteria were renal failure, polymyositis and conservative fracture management. Troponin T levels were measured on admission, day 1 and 2 post-surgery. According to local protocol, a level of >0.03 ng/mL was considered to be raised. Adverse outcome measures were cardiorespiratory events (myocardial infarction, congestive cardiac failure, unstable angina, major arrhythmias requiring treatment and pulmonary embolism), death and length of inpatient stay. RESULTS: One hundred and twenty-nine patients presented with femoral neck fractures. 108 patients were included after application of the exclusion criteria. 42 (39%) showed a Troponin rise. Of these, 25 sustained one or more outcome complications versus seven with no rise (p<0.001). The mean hospital stay was 25.7 days for patients with elevated Troponin, 18.3 days in the normal group (p<0.012). There were nine deaths in the raised Troponin group, and five with no rise (p<0.05). DISCUSSION: The principle causes of early death after hip fracture surgery are cardiac failure and myocardial infarction. Troponin T is a sensitive enzymatic marker of myocardial injury. The association between raised Troponin and hip fractures has not previously been made. In our series, 39% showed a perioperative Troponin rise. This was significantly associated with increased morbidity, mortality and longer hospitalisation. Many patients appear to be having silent events, causing significant morbidity. We recommend Troponin measurement in all patients to identify this risk and allow appropriate optimisation measures. PMID- 18407278 TI - In vitro simulation and quantification of wear within the patellofemoral joint replacement. AB - Quantification of the wear rate in vitro is now considered an essential step in the development of a new joint replacement prior to clinical trials. However, little research exists around in vitro simulation of wear in the patellofemoral joint (PFJ) despite over 200,000 being implanted annually within the European Union. A method to simulate wear in the laboratory using four input degrees of freedom within the PFJ of total knee replacement (TKR) has been developed. Wear simulation was validated through comparison of functional kinematics and patellar surface damage modes produced in vitro to clinical outcomes. The technique has been shown to replicate the prescribed in vivo kinematics in a reproducible and repeatable manner. The wear scar areas were similar to those found in vivo. However, geometrical measurements of wear were not reliable due to creep and geometry changes. As has been found previously with tibial inserts, geometrical determination of wear volume was not found to be an effective method of comparing wear from simulators and retrievals. Change in volume calculated gravimetrically was seen to be the most repeatable measure of patellar wear in vitro. PMID- 18407279 TI - Simple preparation of baicalin from Scutellariae Radix. AB - A simple, rapid and reproducible method was developed to isolate high-purity baicalin from Scutellariae Radix, the dried roots of Scutellaria baicalensis Georgi. The method involves partition/recrystallization steps without repeated column chromatography or special instruments. Isolated baicalin was characterized by comparisons of TLC, HPLC, IR, MS, and NMR data with an authentic sample. Moreover, beta-glucuronidase hydrolysis of baicalin sequentially yielded glucuronic acid and baicalein as confirmed by co-TLC with authentic samples. The purity of baicalin was more than 97% with yield ca. 8.7% (w/w). The method presented here appears suitable for commercial application. PMID- 18407280 TI - Capillary liquid chromatography-mass spectrometry for the rapid identification and quantification of almond flavonoids. AB - A rapid negative ion ESI high-performance capillary liquid chromatography-mass spectrometry method was developed to identify and quantify flavonoids (e.g., flavanols, flavonols, flavanones and glycosides). Fifteen standards and two varieties of almond skin extract powder (Carmel and Nonpareil) were used to demonstrate the chromatographic separation, reproducibility and accuracy of the method that employed a 150 mm x 0.3 mm ChromXP 3C18-EP-120 column. All standards eluted in less than 10 min, providing a 9-12x reduction in analysis time compared to existing methods (90-120 min). However, isomers (e.g., catechin/epicatechin and galactosides/glucosides) were not resolved and, therefore, identified and quantified collectively. RSDs for retention time and peak area reproducibility (mass spectrometry data) were <0.5% and <5.0%, respectively. Peak area reproducibility was greatly improved (from a RSD>10%) after the implementation of a low-flow metal needle in the ESI source. Quantitation by mass spectrometry also afforded a % error less than 5% for most compounds. PMID- 18407281 TI - Micro-proteome analysis using micro-chromatofocusing in intact protein separations. AB - Multi-dimensional liquid-based separation is required for fractionation and mapping of complex protein mixtures from cells. A method that has been used as the first dimension in such separations is chromatofocusing (CF), which is based on generating a pH gradient on an anion exchange column. The use of pH in the first dimension is essential where pH is a fundamental property of proteins and can provide information on post-translationally modified forms of a protein. In this work, a micro-chromatofocusing technique is introduced which can separate microgram levels of proteins from cell lysates for further analysis by LC-MS/MS. It is shown that this method can analyze 10 microg of sample and detect nearly 700-800 proteins from ovarian cancer cell line lysates. PMID- 18407282 TI - Novel wavelength-resolved fluorescence detection for a high-throughput capillary electrophoresis system under a diascopic configuration. AB - This paper presents a novel method regarding a wavelength-resolved fluorescence detection scheme for high-throughput analysis of bio-samples in a micro-CE chip. Instead of using the conventional laser-induced fluorescence (LIF) microscope equipped with delicate spatial filters and complex control systems, this study adopts a hollow cone illumination generated using a dark-field condenser for exciting fluorescence in the microchannel and an ultraviolet-visible-near infrared (UV-Vis-NIR) spectrometer for detecting the emission signals. Experimental results show that the proposed system is feasible for simultaneously detecting a mixed sample composed of Atto 610, Rhodamine B and fluorescein isothiocyanate (FITC) fluorescent dyes in a single test run. Furthermore, a mixed bio-sample composed of two mixed 16-mer single-stranded DNAs labeled with Cy3 and FITC fluorescent dyes is also successfully detected with the proposed system. The measured limit of detection (LOD) for detecting FITC of the proposed system can be as low as 5.4x10(-6)M (S/N=3). This proposed detection method has shown its potential on RNA identification and DNA sequencing applications. PMID- 18407283 TI - Adsorption and surface complexation of trimesic acid at the alpha-alumina electrolyte interface. AB - Adsorption kinetics, adsorption isotherms and surface complexation of trimesic acid onto alpha-alumina surfaces were investigated. Adsorption kinetics of trimesic acid with an initial concentration of 0.5 mM onto alpha-alumina surfaces were carried out in batch method in presence of 0.05 mM NaCl (aq) at pH 6 and 298.15, 303.15 and 313.15 K. Adsorption isotherms were carried out at 298.15 K, pH 5-9, and 0.05 mM NaCl (aq) by varying trimesic acid concentration from 0.01 to 0.6 mM. Three kinetics equations such as pseudo-first-order, pseudo-second-order and Ho equations were used to estimate the kinetics parameters of the adsorption of trimesic acid on the alpha-alumina surfaces. Ho equation fits the experimental kinetics data significantly better and the estimated equilibrium concentration is in excellent agreement with the experimental value. The adsorption data were fitted to Freundlich and Langmuir adsorption model and the later best fits the adsorption isotherms. Comparison of adsorption density of trimesic acid with that of benzoic and phthalic acids follows the sequence: benzoic acid < trimesic acid < phthalic acid. The negative activation energy and the Gibbs free energy for adsorption indicate that the adsorption of trimesic acid onto alpha-alumina is spontaneous and facile. DRIFT spectroscopic studies reveal that trimesate forms outer-sphere complexes with the surface hydroxyl groups that are generated onto alpha-alumina surfaces in the pH range of the study. PMID- 18407284 TI - Optimization of instantaneous solvent exchange/surface modification process for ambient synthesis of monolithic silica aerogels. AB - The instantaneous solvent exchange/surface modification (ISE/SM) process for the ambient synthesis of crack-free silica aerogel monoliths with a high production yield was optimized. Monolithic forms of silica wet gels were obtained from aqueous colloidal silica sols prepared via the ion exchange of sodium silicate solutions. Crack-free silica aerogel monoliths were synthesized via an ISE/SM process using isopropyl alcohol/trimethylchlorosilane as a modification agent and n-hexane as a main solvent, followed by ambient drying. The optimum process conditions of the ISE/SM process were investigated by clarifying the reaction mechanism and phenomena. Most effective ranges of process variables on the ISE/SM stage were determined as 0.2500-0.3567 of TMCS/H2O (pore water) in molar ratio and 15-30 of n-hexane/TMCS in volumetric ratio, with a reaction temperature below 283 K. Crack-free silica aerogel monoliths synthesized via these conditions had a well-developed mesoporous structure and excellent properties (bulk density of 0.12-0.14 g/cm3, specific surface area of 724 m2/g), and a high yield (nearly 80%). PMID- 18407285 TI - Light-sensitive lamellar phases. AB - Light sensitive lamellar (L(alpha)) phases have been generated in glycerol/water mixtures from blends of an inert ABA tri-block co-polymer (dimethylsiloxane polyethylenoxide, (EO)(15)-(PDMS)(15)-(EO)(15)), and a photodestructible anionic surfactant C(6)PAS (sodium 4-hexylphenylazosulfonate). These L(alpha) systems have been formulated in a 6/4 glycerol/water (v/v) mixed solvent. Rheology, small angle X-ray scattering (SAXS), small-angle neutron scattering (SANS) and dynamic light scattering (DLS) have been used to characterize the change in phase behavior and structure after the incorporated C(6)PAS is selectively degraded by UV light incident on the L(alpha) phases. The ABA co-polymer alone forms weakly structured lamellar mesophases, which are stabilized by thermal fluctuations and characterized by low shear moduli (L(alpha) phase-A). Addition of C(6)PAS (10 mM) introduces charge stabilization, generating more ordered and stiffer L(alpha) systems (L(alpha) phase-B). After UV irradiation (Hg lamp) of the L(alpha)-B phases, and subsequent degradation of the ionic C(6)PAS, the results of rheological and scattering studies are consistent with an irreversible change back to L(alpha) phase-A type systems. These formulations display a novel transition between electrostatically- and thermally-stabilized lamellae, which may be controlled by incident UV light. PMID- 18407286 TI - Lexical priming of function words and content words with children who do, and do not, stutter. AB - The specific mechanisms that underlie childhood stuttering are not fully understood. The current study investigated these mechanisms by comparing the effect on fluency of priming different components of a short sentence. The main findings were that: (1) both children who stutter (CWS) (n=12, M age=6;3) and children who do not stutter (CWNS) (n=12, M age=6;6) were more fluent after function word (FW) priming than content word (CW) priming, (2) this effect was significantly greater for CWS than for CWNS, and (3) after FW priming, CWS produced CWs with significantly longer duration than did CWNS. These findings are discussed in relation to two competing theories of stuttering: the covert repair hypothesis (CRH) [Kolk, H., & Postma, A. (1997). Stuttering as a covert repair phenomenon. In R. F. Curlee & G. M. Siegel (Eds.), Nature and treatments of stuttering: New directions (pp. 182-203). Needham Heights, MA: Allyn & Bacon] and the developmentally focused model of Howell and Au-Yeung [Howell, P., & Au-Yeung, J. (2002). The EXPLAN theory of fluency control and the diagnosis of stuttering. In E. Fava (Ed.), Current issues in linguistic theory series: Pathology and therapy of speech disorders (pp. 75-94). Amsterdam: John Benjamins]. LEARNING OUTCOMES: After reading this article, the reader will be able to: (1) understand which linguistic levels can be primed in children who stutter; (2) see why EXPLAN predicts asymmetrical effects on fluency when function or content words are primed; (3) appreciate the distinguishing characteristics of CRH and EXPLAN theories. PMID- 18407287 TI - Development and validation of a multiplex immunoassay for the simultaneous determination of serum antibodies to Bordetella pertussis, diphtheria and tetanus. AB - To increase testing of vaccine induced humoral immunity in immune surveillance studies and vaccine trials, a rapid and simple microsphere-based multiplex assay (pentaplex) was developed for the quantitation of IgG serum antibodies directed against the Bordetella pertussis antigens: Pertussis Toxin (Ptx), Filamentous hemagglutinin (FHA), Pertactin (Prn) and to Diphtheria toxin and Tetanus toxin. All individual antigens were covalently linked to carboxylated microspheres. The method was validated with different serum panels (n=60-78 samples). With the Multiplex Immunoassay (MIA) no evidence for bead interference between monoplex and pentaplex was found. The specificity of the method was shown by a heterologous inhibition of <16% and homologous inhibition of >92%. The pentaplex MIA appeared sensitive with lower limits of quantitation (LLOQ) well below those for ELISA (enzyme-linked immuno-sorbant assay). Assay reproducibility was high with intra-assay variability less than 10% and inter-assay variability below 14%. The reproducibility of the bead conjugation was good and beads could be stored up to at least 6 months without quality reduction. Importantly, the correlation of the pentaplex MIA with the individual ELISAs was excellent, R>0.98 for the Pertussis antigens and R=0.95 for Diphtheria and R=0.98 for Tetanus. Serum IgG antibodies to B. pertussis, Diphtheria and Tetanus can be measured easily, specific and reproducible using the pentaplex MIA. The pentaplex MIA shares features of the ELISA with the additional advantages of high sample throughput and small sample volumes and antigen required. PMID- 18407288 TI - PCR-based method for the detection of cry genes in local isolates of Bacillus thuringiensis from Thailand. AB - A total of 134 isolates of Bacillus thuringiensis obtained from different geographical and ecological origins in Thailand were analyzed to determine the distribution and diversity of cry1, cry2 and cry9 genes encoding for Cry proteins toxic to lepidopteran insects. Strains containing cry1-type genes (109/134 or 81.3%) were found at the same frequency as strains harboring cry2 gene (108/134 or 80.6%) whereas only 50 strains contained cry9 gene (50/134 or 37.3%). Seventeen percent (23/134) of the B. thuringiensis isolates did not harbor any cry1, cry2 or cry9 genes. Among cry1 containing isolates, cry1A (49.3%), cry1B (50.0%), cry1G (48.5%), cry1I (49.3%), cry1J (35.1%) and cry1L (47.0%) were considered abundant. The cry2 gene was distributed with high frequency (>70%) in every region of the country. The study of cry gene combinations revealed 14 cry gene profiles. PMID- 18407289 TI - Patterns of p53, p73 and mortalin gene expression associated with haemocyte polyploidy in the soft-shell clam, Mya arenaria. AB - The molecular mechanisms by which haemocytes of clams are transformed in the course of haemic neoplasia remain by far unknown. The aim of this study was to quantify the expression of p53/p73 and mortalin genes, in relation with the ploidy status of clam haemocytes and to correlate the p53 expression with mortalin expression. For this purpose, soft-shell clams, Mya arenaria, were collected from an endemic zone for neoplasia. The ploidy of haemocytes was assessed for each individual clam by flow cytometry using a propidium iodide protocol, while p53/p73 and mortalin gene expressions were quantified by real time RT-PCR. Results show that haemocytes of some clams with a moderate percentage (15-50%) of tetraploid cells have a significantly high level of p53 and p73 in comparison with clams belonging to categories with low (<15%) or high levels (>50%) of tetraploid cells, where low levels of expression of these genes were observed. Furthermore, mortalin gene expression is strongly correlated (r(2)=0.68, p<0.01) with p53 gene expression level. This reinforces the hypothesis of a cytoplasmic p53 sequestration mechanism in clam haemic neoplasia. Further studies are needed to confirm these preliminary results and further unravel the molecular pathways involved in this process. Our results are believed to provide phenotypic foundation for such studies to be undertaken. PMID- 18407291 TI - Novel structural and functional mode of a knot essential for RNA binding activity of the Esa1 presumed chromodomain. AB - Chromodomains are methylated histone binding modules that have been widely studied. Interestingly, some chromodomains are reported to bind to RNA and/or DNA, although the molecular basis of their RNA/DNA interactions has not been solved. Here we propose a novel binding mode for chromodomain-RNA interactions. Essential Sas-related acetyltransferase 1 (Esa1) contains a presumed chromodomain in addition to a histone acetyltransferase domain. We initially determined the solution structure of the Esa1 presumed chromodomain and showed it to consist of a well-folded structure containing a five-stranded beta-barrel similar to the tudor domain rather than the canonical chromodomain. Furthermore, the domain showed no RNA/DNA binding ability. Because the N-terminus of the protein forms a helical turn, we prepared an N-terminally extended construct, which we surprisingly found to bind to poly(U) and to be critical for in vivo function. This extended protein contains an additional beta-sheet that acts as a knot for the tudor domain and binds to oligo(U) and oligo(C) with greater affinity compared with other oligo-RNAs and DNAs examined thus far. The knot does not cause a global change in the core structure but induces a well-defined loop in the tudor domain itself, which is responsible for RNA binding. We made 47 point mutants in an esa1 mutant gene in yeast in which amino acids of the Esa1 knotted tudor domain were substituted to alanine residues and their functional abilities were examined. Interestingly, the knotted tudor domain mutations that were lethal to the yeast lost poly(U) binding ability. Amino acids that are related to RNA interaction sites, as revealed by both NMR and affinity binding experiments, are found to be important in vivo. These findings are the first demonstration of how the novel structure of the knotted tudor domain impacts on RNA binding and how this influences in vivo function. PMID- 18407290 TI - Taurine depletion caused by knocking out the taurine transporter gene leads to cardiomyopathy with cardiac atrophy. AB - The sulfur-containing beta-amino acid, taurine, is the most abundant free amino acid in cardiac and skeletal muscle. Although its physiological function has not been established, it is thought to play an important role in ion movement, calcium handling, osmoregulation and cytoprotection. To begin examining the physiological function of taurine, we generated taurine transporter- (TauT-) knockout mice (TauTKO), which exhibited a deficiency in myocardial and skeletal muscle taurine content compared with their wild-type littermates. The TauTKO heart underwent ventricular remodeling, characterized by reductions in ventricular wall thickness and cardiac atrophy accompanied with the smaller cardiomyocytes. Associated with the structural changes in the heart was a reduction in cardiac output and increased expression of heart cardiac failure (fetal) marker genes, such as ANP, BNP and beta-MHC. Moreover, ultrastructural damage to the myofilaments and mitochondria was observed. Further, the skeletal muscle of the TauTKO mice also exhibited decreased cell volume, structural defects and a reduction of exercise endurance capacity. Importantly, the expression of Hsp70, ATA2 and S100A4, which are upregulated by osmotic stress, was elevated in both heart and skeletal muscle of the TauTKO mice. Taurine depletion causes cardiomyocyte atrophy, mitochondrial and myofiber damage and cardiac dysfunction, effects likely related to the actions of taurine. Our data suggest that multiple actions of taurine, including osmoregulation, regulation of mitochondrial protein expression and inhibition of apoptosis, collectively ensure proper maintenance of cardiac and skeletal muscular structure and function. PMID- 18407292 TI - Complexity does not affect stability in feasible model communities. AB - The complexity-stability relation is a central issue in ecology. In this paper, we show how the sampling method most often used to parameterize an ecological community, can affect the conclusions about whether or not complexity promotes stability and we suggest a sampling algorithm that overcomes the problem. We also illustrate the importance of treating feasibility separately from stability when constructing model communities. Using model Lotka-Volterra competition communities we found that probability of feasibility decreases with increasing interaction strength and number of species in the community. However, for feasible systems we found that local stability probability and resilience do not significantly differ between communities with few or many species, in contrast with earlier studies that, did not account for feasibility and concluded that species-poor communities had higher probability of being locally stable than species-rich communities. PMID- 18407293 TI - Modeling absence seizure dynamics: implications for basic mechanisms and measurement of thalamocortical and corticothalamic latencies. AB - A successful physiologically based continuum model of the corticothalamic system is applied to determine the relative contributions of axonal and intrinsic cellular delays to the waveforms of absence seizures. The predicted period of the absence seizure depends linearly on model parameters describing thalamocortical, corticothalamic, intracortical, and synaptodendritic delays, and these dependences are linked to the seizure mechanism by showing how time intervals between peaks in the waveforms depend on the parameters. Counterintuitively, it is found that a peak in the local field potential recorded in the thalamic relay nuclei can precede the peak in the cortical field that drove it, without violating causality, but rendering naive interpretation of time intervals between peaks invalid. We argue that a thalamocortical loop mechanism for absence seizures is consistent with intrathalamic cellular properties being the leading determinant of the frequency of spike-wave discharges in rat genetic models, with the combination of network and cellular properties providing a natural explanation for the lower frequency of human absence seizures. Finally, our results imply that the seizure frequency is not determined by the fastest thalamocortical and corticothalamic fibers, but rather depends on an effective weighted conduction velocity of all pathways present. PMID- 18407294 TI - The evolution of fidelity in sensory systems. AB - We investigate the effect that noise has on the evolution of measurement strategies and competition in populations of organisms with sensory systems of differing fidelities. We address two questions motivated by experimental and theoretical work on sensory systems in noisy environments: (1) How complex must a sensory system be in order to face the need to develop adaptive measurement strategies that change depending on the noise level? (2) Does the principle of competitive exclusion for sensory systems force one population to win out over all others? We find that the answer to the first question is that even very simple sensory systems will need to change measurement strategies depending on the amount of noise in the environment. Interestingly, the answer to the second question is that, in general, at most two populations with different fidelity sensory systems may co-exist within a single environment. PMID- 18407295 TI - Ex vivo expansions of megakaryocytopoiesis from placental and umbilical cord blood CD34(+) cells in serum-free culture supplemented with proteoglycans extracted from the nasal cartilage of salmon heads and the nasal septum cartilage of whale. AB - As a possible approach to the treatment of thrombopocytopenia, the ex vivo expansion of megakaryocytic progenitor cells may be a useful tool to accelerate platelet recovery in vivo. Our objective was to assess the promoting effect of proteoglycans in a serum-free culture condition using human cord blood CD34(+) cells. Highly purified proteoglycan (PG) extracted from the nasal cartilage of salmon heads and the nasal septum cartilage of a whale were applied to the ex vivo expansion of megakaryocytopoiesis and thrombopoiesis from placental and umbilical cord blood CD34(+) cells in serum-free cultures stimulated with a combination of thrombopoietin (TPO) and interleukin-3 (IL-3). Each PG (0.5 and 5 mug) was applied to the culture with three different concentrations of TPO (50, 5 and 0.5 ng/ml) and IL-3 (100, 10 and 1 ng/ml). Both of the PGs showed no promoting effects on the mononuclear cell proliferation rate in any of the cultures. However, the whale-PG promoted the generation of megakaryocytic progenitor cells and megakaryocytes in the culture with a lower dose of cytokines, respectively. In addition, whale-PG led to a significant increase in CD42a(+) particles which seemed to be platelets. While the salmon-PG failed to promote such production in almost all of the cultures. Although whale-PG is an attractive molecule for the ex vivo expansion of human megakaryocytopoiesis, its action may depend on the glycosaminoglycans sulfation pattern and the ability of the binding affinity and the kinetics to interact with the cytokines and hematopoietic stem/progenitor cells. PMID- 18407296 TI - (-)-Epigallocatechin gallate reduces transforming growth factor beta-stimulated HSP27 induction through the suppression of stress-activated protein kinase/c-Jun N-terminal kinase in osteoblasts. AB - We previously reported that transforming growth factor-beta (TGF-beta) stimulates heat shock protein 27 (HSP27) induction through p38 mitogen-activated protein (MAP) kinase and extracellular signal-regulated kinase 1/2 (ERK1/2) in osteoblast like MC3T3-E1 cells. In the present study, we investigated whether (-) epigallocatechin gallate (EGCG), the major polyphenol found in green tea, affects the TGF-beta-stimulated induction of HSP27 in these cells, and its underlying mechanism. EGCG significantly suppressed the HSP27 induction stimulated by TGF beta in a dose-dependent manner between 10 and 30 microM without affecting the HSP70 levels. TGF-beta with or without EGCG did not affect the advanced oxidation protein products. The TGF-beta-induced phosphorylation of p38 MAP kinase and ERK1/2 was not affected by EGCG. SP600125, a specific inhibitor of stress activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), markedly reduced the HSP27 expression induced by TGF-beta. EGCG significantly suppressed the TGF beta-induced phosphorylation of SAPK/JNK without affecting the phosphorylation of Smad2. EGCG attenuated the phosphorylation of both MKK4 and TAK1 induced by TGF beta. These results strongly suggest that EGCG suppresses the TGF-beta-stimulated induction of HSP27 via the attenuation of the SAPK/JNK pathway in osteoblasts, and that this effect is exerted at a point upstream from TAK1. PMID- 18407297 TI - Molecular characterization and expression pattern of taurine transporter in zebrafish during embryogenesis. AB - Taurine and its transporter (TauT) are expressed in preimplantation embryos, but their role in embryogenesis is not known. To investigate the role of TauT during embryonic development, we cloned and functionally characterized the zebrafish TauT. The zebrafish TauT cDNA codes for a protein of 625 amino acids which is highly homologous to mammalian TauT. When expressed in mammalian cells, zebrafish TauT mediates taurine uptake in a Na(+)/Cl(-)-dependent manner with a Na(+):Cl( ):taurine stoichiometry of 2:1:1. In the zebrafish embryo, taurine and TauT mRNA are present during early cleavage stages, indicating that both the transporter and its substrate are maternally derived. During embryogenesis, zygotic expression of TauT mRNA is evident in the retina, brain, heart, kidney, and blood vessels. Knockdown of TauT by antisense morpholino oligonucleotides leads to cell death in the central nervous system and increased mortality. These findings suggest a specific role for TauT during development in vertebrates. PMID- 18407298 TI - Organochlorine pesticides in fish from the southern Baltic Sea: levels, bioaccumulation features and temporal trends during the 1995-2006 period. AB - The levels of HCH isomers, HCB and summation DDTs were determined in five species of Baltic fish collected during 1995-2006. Some downward time trends of HCH isomer, pp'-DDE and pp'-DDD concentrations in Baltic fish were observed; in contrast, HCB and DDT concentrations did not exhibit any obvious trend. Concentrations of summation HCHs expressed on a lipid weight basis were very similar in all species studied. In contrast, concentrations of summation DDTs varied in tested species. The most abundant HCH isomer was beta-HCH and among DDT related compounds, pp'-DDE prevailed. In our investigations the sampling sites were not a crucial factor for organochlorine pesticide concentrations and patterns observed in fish, but the occurrence of several inter-species differences in the bioaccumulation features of OC pesticides were observed. For cod there was a positive correlation between fish size (length) and summation DDT concentration. In herring, sprat and salmon samples, summation DDT concentrations were negatively correlated with fat content. PMID- 18407299 TI - Inhibition of NMDA-gated ion channels by bis(7)-tacrine: whole-cell and single channel studies. AB - Bis(7)-tacrine is a novel dimeric acetylcholinesterase inhibitor derived from tacrine, and has been proposed as a promising agent to treat Alzheimer's disease. We have recently reported that bis(7)-tacrine prevents glutamate-induced neuronal apoptosis by antagonizing NMDA receptors. The purpose of this study was to characterize bis(7)-tacrine inhibition of NMDA-activated current by using patch clamp recording techniques. In cultured rat hippocampal neurons, bis(7)-tacrine inhibited NMDA-activated whole-cell current in a concentration-dependent manner with an IC(50) of 0.66+/-0.07 microM. Bis(7)-tacrine produced a gradual decline of NMDA-activated current to a steady-state, but this was not an indication of use-dependence. Also, the slow onset of inhibition by bis(7)-tacrine was not apparently due to an action at an intracellular site. Bis(7)-tacrine, 0.5 microM, decreased the maximal response to NMDA by 40% without changing its EC(50). Bis(7) tacrine inhibition of NMDA-activated current was not voltage-dependent, and was independent of glycine concentration. Results of single-channel experiments obtained from cells expressing NR1 and NR2A subunits revealed that bis(7)-tacrine decreased the open probability and frequency of channel opening, but did not significantly alter the mean open time or introduce rapid closures. These results suggest that bis(7)-tacrine can inhibit NMDA receptor function in a manner that is slow in onset and offset and noncompetitive with respect to both NMDA and glycine. The noncompetitive inhibition of NMDA receptors by bis(7)-tacrine could contribute to its protective effect against glutamate-induced neurotoxicity. PMID- 18407300 TI - The neural correlates of change detection in the face perception network. AB - A common view is that visual processing within the ventral visual stream is modulated by attention and awareness. We used fMRI adaptation to investigate whether activation in a network of brain regions involved with face recognition- namely the fusiform face area (FFA), occipital face area (OFA) and right superior temporal sulcus (rSTS)--was modulated by physical changes to face stimuli or by observers' awareness of the changes. We sequentially presented two matrices of four faces. In two thirds of the trials one of the faces changed. We compared activations generated in three conditions (i) change detected trials, (ii) change blind trials, and (iii) no change trials. If face areas were sensitive to physical changes then we expected similar levels of activation for face changes regardless of change detection. If face areas were sensitive to levels of awareness of change then we expected greater levels of activation for detected changes compared to no change detection. We found that all three-face regions recovered from adaptation when subjects were aware of changes, but only OFA recovered from adaptation when subjects were not aware of the changes. These results suggest that within the face network OFA is involved in representing information that has not yet entered awareness and that consciousness is not an all-or-none phenomenon throughout the ventral stream. PMID- 18407301 TI - Defective emotion recognition in early HD is neuropsychologically and anatomically generic. AB - Huntington's disease (HD) is an inherited neurodegenerative disorder that classically presents with motor, cognitive and psychiatric symptoms. However, other abnormalities also occur in this condition, notably deficient recognition of facial emotional expressions. Deficits in emotion recognition impact significantly on the lives of HD patients and their families and thus it is important to clarify the onset and pattern of impairment. This study investigated facial emotion recognition in a large cohort of early HD patients, and premanifest gene-carriers. We used voxel-based morphometry (VBM) to examine the neuroanatomical correlates of emotion recognition performance. Forty patients with early HD, 21 premanifest gene carriers and 20 controls were assessed using 24 faces from the Ekman Pictures of Facial Affect, and volumetric brain MRI. The HD group was significantly worse than controls at recognising, surprise, disgust, anger and fear, and worse than the premanifest group at recognising disgust and anger. When patient data were expressed as z-scores, recognition of anger was significantly worse than disgust in the early HD group. In the VBM analysis, these deficits were associated with common regional atrophy: impaired recognition of surprise, disgust, anger and fear were all associated with striatal volume loss. Fear was associated with additional atrophy of the right insula and left and right lateral orbitofrontal cortex. Even in early HD there is a wide-ranging impairment in recognition of negative emotions denoting 'threat'. Our findings implicate a generic fronto-subcortical network in the pathogenesis of these emotion recognition deficits. PMID- 18407302 TI - Matching boxes: familiar size influences action programming. AB - The perception/action model is the dominant account of the primary division of labour in the human visual pathway. Integral to this model is the idea that goal directed actions are guided spatially by bottom-up vision, independent of perceptual recognition and top-down object knowledge. We question this idea by showing that the expected size of familiar objects (matchboxes) affects the amplitude of reaches made to grasp them, and the pre-shaping of the hand, even when binocular cues are available. This suggests that perceptual recognition routinely influences action programming. PMID- 18407303 TI - Hypertension in Orissa, India: a cross-sectional study among some tribal, rural and urban populations. PMID- 18407304 TI - Application of PCR-RFLP for the exploration of the molecular diversity of Leishmania infantum in Algeria. AB - In recent years, new methods have been developed for the molecular typing of Leishmania that need to be extensively validated by studies of clinical isolates in a well defined epidemiological context. The present study is a contribution to this effort. Using PCR-RFLP of gp63 and cpb genes, we analysed 59 isolates of L. (L.) infantum obtained from different regions of Algeria and originating from different clinical forms, hosts and zymodemes. PCR-RFLP identified 15 different genotypes among the four zymodemes analysed, thereby demonstrating a higher discriminatory power than multilocus enzyme electrophoresis. We did not see any significant relationships between PCR-RFLP patterns and host origin. However, cpb polymorphism showed two interesting trends: a possible relationship with the cutaneous origin of the isolates and an association with a West-East cline. We verified the proof of evidence of the direct applicability of gp63 and cpb PCR RFLP in blood samples from dogs. Further work is needed to compare the sensitivity of pattern detection with cpb and gp63 PCR-RFLP but our results pave the way to future multilocus PCR-RFLP studies of L. (L.) infantum populations. PMID- 18407305 TI - Egg envelope synthesis and chorion modification after oviposition in the dragonfly Libellula depressa (Odonata, Libellulidae). AB - Libellula depressa (Odonata, Libellulidae) is an exophytic dragonfly ovidepositing eggs in clutches on the surface of floating plants and algae. The present work investigates, at ultrastructural level, the gradual differentiation of the egg envelopes and the chorionic changes after egg deposition in water. The ovary of the mature female of L. depressa is composed of numerous strings of panoistic ovarioles, where the eggshell formation takes place gradually throughout the activity of the follicle cells. The present data show that the egg envelopes are constituted of a very thick electrondense vitelline envelope, a thin endochorion and an extremely thick exochorion composed of a fibrillar matrix resting on a thin electrondense layer. After deposition in water, L. depressa eggs, initially white and almost transparent, gradually become brown spots in a semitransparent jelly coat, rich of incorporated debris. The jelly coat enveloping the eggs of L. depressa derives exclusively from the exochorion, constituted of a fibrillar matrix, which swell at contact with water. The jelly like coat performs an adhesive function and presumably a protective role during egg segmentation and ensuing larval hatching. PMID- 18407306 TI - Developmental toxicity and alteration of gene expression in zebrafish embryos exposed to PFOS. AB - Perfluorooctanesulfonate (PFOS) is a persistent organic pollutant, the potential toxicity of which is causing great concern. In the present study, we employed zebrafish embryos to investigate the developmental toxicity of this compound. Four-hour post-fertilization (hpf) zebrafish embryos were exposed to 0.1, 0.5, 1, 3 and 5 mg/L PFOS. Hatching was delayed and hatching rates as well as larval survivorship were significantly reduced after the embryos were exposed to 1, 3 and 5 mg/L PFOS until 132 hpf. The fry displayed gross developmental malformations, including epiboly deformities, hypopigmentation, yolk sac edema, tail and heart malformations and spinal curvature upon exposure to PFOS concentrations of 1 mg/L or greater. Growth (body length) was significantly reduced in the 3 and 5 mg/L PFOS-treated groups. To test whether developmental malformation was mediated via apoptosis, flow cytometry analysis of DNA content, acridine orange staining and TUNEL assay was used. These techniques indicated that more apoptotic cells were present in the PFOS-treated embryos than in the control embryos. Certain genes related to cell apoptosis, p53 and Bax, were both significantly up-regulated upon exposure to all the concentrations tested. In addition, we investigated the effects of PFOS on marker genes related to early thyroid development (hhex and pax8) and genes regulating the balance of androgens and estrogens (cyp19a and cyp19b). For thyroid development, the expression of hhex was significantly up-regulated at all concentrations tested, whereas pax8 expression was significantly up-regulated only upon exposure to lower concentrations of PFOS (0.1, 0.5, 1 mg/L). The expression of cyp19a and of cyp19b was significantly down-regulated at all exposure concentrations. The overall results indicated that zebrafish embryos constitute a reliable model for testing the developmental toxicity of PFOS, and the gene expression patterns in the embryos were able to reveal some potential mechanisms of developmental toxicity. PMID- 18407307 TI - Arsenic interferes with the signaling transduction pathway of T cell receptor activation by increasing basal and induced phosphorylation of Lck and Fyn in spleen cells. AB - Arsenic is known to produce inhibition as well as induction of immune cells proliferative responses depending on the doses as one of its mechanisms of immunotoxicity. Here we evaluate the effect of arsenic exposure on the activation of splenic mononuclear cells (SMC) in male CD57BL6N mice. Intra-gastric exposure to arsenic (as sodium arsenite) for 30 days (1, 0.1, or 0.01 mg/kg/day), reduced the proportion of CD4+ cells and the CD4+/CD8+ ratio in the spleen, increasing the proportion of CD11b+ cells. Arsenic exposure did not modify the proportion of B cells. SMC showed an increased level of phosphorylation of lck and fyn kinases (first kinases associated to TCR complex when activated). Although normal levels of apoptosis were observed on freshly isolated SMC, an increase in apoptotic cells related with the increase in phosphorylation of lck and fyn was observed when SMC were activated with Concanavalin-A (Con-A). Arsenic exposure reduced the proliferative response of SMC to Con-A, and also reduced secretion of IL-2, IL-6, IL-12 and IFNgamma. No effect was observed on IL-4, and IL-10 secretion. The same effects were observed when SMC of exposed animals were activated with anti CD3/CD28 antibodies for 24 h, but these effects were transitory since a recovery, up to control levels or even higher, were observed after 72 h of stimulation. This study demonstrates that repeated and prolonged exposure to arsenic alters cell populations and produces functional changes depending on the specific activation pathway, and could be related with the phosphorylation status of lck and fyn kinases. PMID- 18407308 TI - An underlying role for hepatobiliary dysfunction in cyclosporine A nephrotoxicity. AB - Renal-derived cysteinyl leukotrienes (cysLT), such as leukotrienes C(4) (LTC(4)) and D(4) (LTD(4)) are thought to mediate acute and chronic cyclosporine A (CSA) nephrotoxicity. However, whole-body cysLT elimination is regulated primarily by hepatobiliary excretion. Since CSA is known to alter hepatobiliary function, the effects of CSA on whole-body cysLT elimination were investigated in vivo, with respect to hepatobiliary and renal function. Male rats were anesthetized and cannulated (jugular vein, bile duct, and urinary bladder). A tracer dose of tritiated LTC(4) ((3)H-LTC(4)) was administered systemically (i.v.) immediately following vehicle and then 90 min later after vehicle or CSA. In vehicle/vehicle controls, hepatobiliary (3)H-cysLT elimination predominated over renal elimination without altering glomerular filtration rate (GFR), bile flow, and urine production. (3)H-cysLT elimination kinetics were comparable between each 90 min collection period. In vehicle/CSA-treated rats, an acutely nephrotoxic dose of CSA (20 mg/kg, i.v.) reduced urine flow 74+/-9% and caused a transient reduction in GFR, while total bile flow decreased 40+/-13%. Hepatobiliary and renal (3)H-cysLT elimination was also impaired 59+/-5 and 61+/-18%, respectively. In contrast, a non-nephrotoxic dose (2 mg/kg i.v.) increased renal (3)H-cysLT elimination due to impaired hepatobiliary elimination without affecting GFR, bile flow or urine production. Both doses caused (3)H-cysLT retention in hepatic and renal tissue. These findings demonstrate that CSA alters whole-body handling of cysLT by disrupting hepatobiliary cysLT elimination. This disruption leads to increased renal exposure to systemically derived cysLT and renal cysLT tissue retention. Renal exposure to and accumulation of systemically derived cysLT products may be underlying factors in CSA nephrotoxicity. PMID- 18407309 TI - Expression of ovarian microsomal epoxide hydrolase and glutathione S-transferase during onset of VCD-induced ovotoxicity in B6C3F(1) mice. AB - 4-vinylcyclohexene diepoxide (VCD) specifically destroys small pre-antral follicles in the rodent ovary. VCD can be detoxified to an inactive tetrol by microsomal epoxide hydrolase (mEH), or by conjugation to glutathione (GSH) by glutathione S-transferase (GST). Formation of VCD-GSH adducts in the mouse ovary 4 h after VCD exposure (0.57 mmol/kg/day) has been demonstrated. Because the mouse ovary expresses both mEH and GST, expression of mEH and GST pi and mu during a time-course of VCD-induced ovotoxicity was evaluated in a neonatal mouse ovarian culture system. Ovaries from postnatal day 4 (PND4) B6C3F(1) mice were incubated with VCD (15 microM) for 2, 4, 6, 8, 10, 12, or 15 days. Following incubation, ovaries were histologically evaluated, or assessed for mRNA or protein expression. VCD did not cause follicle loss (p>0.05) on days 2, 4, or 6 of culture. At days 8, 10, 12, and 15, VCD reduced (p<0.05) both primordial and primary follicle numbers. Increased (p<0.05) expression of mEH, GST pi and GST mu mRNA was detected after 4 days of VCD exposure. This expression was reduced on days 6 and 8, when follicle loss was underway, but increased (p<0.05) after 10 days of exposure. mEH and GST pi proteins were elevated (p<0.05) following 8 days of VCD-exposure however there was no increase in GST mu protein. These findings suggest that with continuous exposure to VCD, increased expression of detoxification enzymes may participate in retarding the onset of follicle loss, but that this loss cannot ultimately be prevented. PMID- 18407310 TI - Effective determination method for a cyanobacterial neurotoxin, beta-N methylamino-L-alanine. AB - We developed a simple and effective analysis procedure that includes pretreatment and determination methods for beta-N-methylamino-l-alanine (BMAA), a cyanobacterial neurotoxin. BMAA may be produced by all known groups of cyanobacteria living in freshwater as well as marine environments. In this paper, we report a novel determination method for BMAA. A cation-exchange resin was effective for the selective concentration of BMAA from cyanobacterial extracts and yielded a high recovery rate. Moreover, liquid chromatography (LC) electrospray ionization mass spectrometry with a hydrophilic LC column was effective for determining BMAA levels. The quantitation limit for BMAA based on selected ion monitoring (SIM) was determined as 0.5 ng at a signal/noise ratio of 5. PMID- 18407311 TI - Expectancies modulate attentional capture by salient color singletons. AB - In singleton feature search for a form-defined target, the presentation of a task irrelevant, but salient singleton color distractor is known to interfere with target detection [Theeuwes, J. (1991). Cross-dimensional perceptual selectivity. Perception & Psychophysics, 50, 184-193; Theeuwes, J. (1992). Perceptual selectivity for color and form. Perception & Psychophysics, 51, 599-606]. The present study was designed to re-examine this effect, by presenting observers with a singleton form target (on each trial) that could be accompanied by a salient) singleton color distractor, with the proportion of distractor to no distractor trials systematically varying across blocks of trials. In addition to RTs, eye movements were recorded in order to examine the mechanisms underlying the distractor interference effect. The results showed that singleton distractors did interfere with target detection only when they were presented on a relatively small (but not on a large) proportion of trials. Overall, the findings suggest that cross-dimensional interference is a covert attention effect, arising from the competition of the target with the distractor for attentional selection [Kumada, T., & Humphreys, G. W. (2002). Cross-dimensional interference and cross trial inhibition. Perception & Psychophysics, 64, 493-503], with the strength of the competition being modulated by observers' (top-down) incentive to suppress the distractor dimension. PMID- 18407312 TI - The transient nature of 2nd-order stereopsis. AB - There are currently two competing dichotomies used to describe how local stereoscopic information is processed by the human visual system. The first is in terms of the type of the spatial filtering operations used to extract relevant image features prior to stereoscopic analysis (i.e. 1st- vs 2nd-order stereo; [Hess, R. F., & Wilcox, L. M. (1994). Linear and non-linear filtering in stereopsis. Vision Research, 34, 2431-2438]). The second is in terms of the temporal properties of the mechanisms used to process stereoscopic information (i.e. sustained vs transient stereo; [Schor, C. M., Edwards, M., & Pope, D. R. (1998). Spatial-frequency and contrast tuning of the transient-stereopsis system. Vision Research, 38(20), 3057-3068]). Here we compare the dynamics of 1st- and 2nd-order stereopsis using several types of stimuli and find a clear dissociation in which 1st-order stimuli exhibit sustained properties while 2nd-order patterns show more transient properties. Our results and analyses unify and simplify two complimentary bodies of work. PMID- 18407313 TI - Judging spatial properties of simple figures. AB - Discrimination was measured for height, area, and aspect ratio of ovals and rectangles. Random jittering of the orthogonal property (width, aspect ratio, and area) was used to control the observers' criterion. Weber fractions for aspect ratio were consistently lower than those for area, and about the same as those for height. Performance with ovals and rectangles did not differ significantly. Two different methods were employed to assess the side effects of jittering. It was found that jittering reduces the discriminability of each property, though less for aspect ratio than for height or area. The hypothesis that judgements of both area and aspect ratio are linear combination of noisy estimates of height and width predicts Weber fractions for aspect ratio and for area to be 2(1/2) times higher than those for height. Results from unjittered trials clearly reject the hypothesis with respect to aspect ratio but not for area. PMID- 18407314 TI - In vitro biotransformation of surfactants in fish. Part I: linear alkylbenzene sulfonate (C12-LAS) and alcohol ethoxylate (C13EO8). AB - Developing regulatory activities (e.g., REACh, [DGEE. 2003. Directorates General Enterprise and Environment. The new EU chemicals legislation REACH. DG Enterprise, Brussels, Belgium. (http://www.europa.eu.int/comm/enterprise/reach/index_en.htm)]) will require bioaccumulation to be assessed for thousands of chemicals. Further, there is increasing pressure to reduce, refine or replace animal tests. Given this scenario, there is an urgent need to evaluate the feasibility of in vitro systems to supply data useful for bioaccumulation estimation. Subcellular and cellular hepatic systems were tested to determine the biotransformation of two surfactants: C12-2-LAS (2-phenyl dodecane p-sulfonate) and an alcohol ethoxylate C13EO8 (Octaethylene glycol monotridecyl ether). The subcellular systems tested were liver homogenates and microsomes from the common carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss). Cellular systems consisted of primary hepatocytes from the common carp (Cyprinus carpio) and PLHC-1 cells, hepatocarcinoma cells from the desert topminnow (Poeciliopsis lucida). All in vitro systems were exposed to radiolabeled test compounds and assayed for biotransformation using liquid scintillation and thin layer chromatographic methods. First-order kinetics were used to estimate rates of biotransformation. Bioconcentration of test materials in fish were predicted using an in vitro to in vivo metabolic rate extrapolation model linked to a mass-balance model commonly used to predict bioaccumulation in fish. Subcellular biotransformation rates for each of the surfactants were greatest with microsomes. Cellular loss rates exceeded subcellular rates, leading to lower predicted BCF values. Predicted BCFs corresponded closely to measured values in several fish species, verifying the utility of in vitro systems in refining Kow-only-based BCFs via the inclusion of biotransformation rates. PMID- 18407315 TI - Serum levels of PCDDs, PCDFs and PCBs in non-occupationally exposed population groups living near two incineration plants in Tuscany, Italy. AB - A pilot study was carried out in Tuscany, Italy, to provide preliminary information on the concentrations of polychlorinated dibenzodioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), dioxin-like polychlorobiphenyls (DL-PCBs), and selected non-dioxin-like PCBs (NDL-PCBs) in groups of subjects living in the vicinity of two incineration plants. Seventy-four volunteers were enrolled from areas identified as under a potential impact from incinerator emissions and from not exposed areas. No significant differences were observed between subjects living in the two types of areas. Total concentrations of PCDDs, PCDFs, and DL PCBs resulted to be in the range 23-30 pg WHO-TEQ g(-1), lipid base, for subjects in the 27-54 year age groups, while concentrations increased to 40-44 pgTEQ g(-1) for the two 55-67 year age groups. The levels of PCDDs and PCDFs were in good agreement with those observed for unexposed population groups in Italy, while the contribution to total TEQ from DL-PCBs was appreciably higher than those currently observed in the general population in Italy and other countries. As to NDL-PCBs, serum levels of the six "indicator" congeners were in the range 240-300 ng g(-1), lipid base, for subjects in the 27-54 year age groups. A raise in NDL PCB body burden (430-470 ng g(-1), lipid base) was observed for the two 55+ year age groups, in agreement with the expected age-dependent increase. The findings from this study do not show an incremental exposure to PCDDs and PCDFs in the samples from subjects living around the two incineration plants, whereas PCB congener profiles in all samples suggest a possible impact on the area of interest of industrial activities from near industrial settlements. PMID- 18407316 TI - Elevated nitrate levels in the groundwater of the Gaza Strip: distribution and sources. AB - Seven years of monitoring groundwater in the Gaza Strip has shown that nitrate was and still is a major groundwater pollutant. The objectives of this research were to study the distribution of NO(3)(-) in the groundwater of the Gaza Strip and to identify the sources of NO(3)(-) in the Gaza aquifer system by assessing nitrogen and oxygen isotopes. The most recent samples collected in 2007 showed 90% of the wells having NO(3)(-) concentrations that are several times higher than the WHO standards of 50 mg/L. Potential NO(3)(-) source materials in Gaza are animal manure N, synthetic NH(4) based fertilizers, and wastewater/sludge. The average concentrations of N in the sludge, manure and soil of Gaza were 2.9%, 1% and 0.08%, respectively. The range in delta(15)N of solid manure samples was +7.5 to +11.9 per thousand. The range in delta(15)N of sludge samples was +4.6 to +7.4 per thousand, while four brands of synthetic fertilizers commonly used in Gaza had delta(15)N ranging from +0.2 to +1.0 per thousand. Sludge amended soil had delta(15)N ranging from +2.0 to +7.3 per thousand. For both delta(18)O and delta(15)N, the ranges of groundwater NO(3)(-) were -0.1 to +9.3 per thousand and +3.2 to 12.8 per thousand, respectively. No significant bacterial denitrification is taking place in the Gaza Strip aquifer. Nitrate was predominantly derived from manure and, provided delta(15)N of sludge represents the maximum delta(15)N of human waste, to a lesser extent from septic effluents/sludge. Synthetic fertilizers were a minor source. PMID- 18407317 TI - Economic benefits of arsenic removal from ground water--a case study from West Bengal, India. AB - People living in almost 50% of the districts in West Bengal are exposed to arsenic contaminated water. This paper seeks to estimate the economic costs imposed by arsenic-related health problems. We use data from a primary survey of 473 households carried out in the districts of North 24 Parganas and Midnapore. We take into account household actions to either decrease the exposure of family members to unsafe water or to alleviate the health effects of consuming arsenic contaminated water. This allows us to assess the benefits of arsenic-safe water by estimating a three equation system that includes averting actions, medical expenditures and a sickness function. We find that by reducing arsenic concentration to the safe limit of 50 microg/l, a representative household will benefit by Rs 297 ($7) per month. The current cost of supplying filtered piped water by the Kolkata Municipal Corporation to households is Rs 127 ($3) per month per household. Thus, investing in safe drinking water is economically feasible and households are willing to pay for such investments if made aware of the effective gain in welfare. Poor households, who make up the highest proportion of arsenic-affected households and incur the largest number of sick days, will be major beneficiaries of such investments. PMID- 18407318 TI - Influence of saliva medium on freeing heavy metal ion from fixed dentures. AB - In dental-prosthetic practice, various kinds of fixed dentures, crowns and bridges, have very often been used in order to replace natural teeth and to respond to all health and esthetic needs. This study investigated the effect of saliva medium on migration of ions of heavy metals from fixed dentures that were fixed with various cements. Also, the influence of saliva medium on natural human teeth was observed. Potentiometric stripping analysis was used in order to determine the content of toxic heavy metals in the examined samples. The study confirmed that synthetic saliva had no significant influence on heavy metal ion migration from the natural teeth, whereas slight migration of some observed toxic heavy metal ions from the fixed dentures was present. This, however, indicates that these contents, although very low, must be taken seriously, because the above mentioned metals have cumulative effect which after some period of time may lead to functional disorders of some organs, and even to some very serious diseases. PMID- 18407319 TI - Lead exposure from the use of Lawsonia inermis (henna) in temporary paint-on tattooing and hair dying. AB - This study reports the evaluation of a number of spectroscopic techniques used in identifying and quantifying the presence of lead in twelve commercial and traditional henna samples. The lead levels found in henna were low with concentrations ranging from 2.29 ppm to 65.98 ppm. Henna is used as a traditional cosmetic and remedy in the Middle East, Far East, and North Africa. The very low concentrations of lead measured in these henna samples were reassuring; however, the cumulative effects of prolonged lead exposure may be of concern. Thus, the use of henna especially among children may constitute a public health risk. PMID- 18407320 TI - Occurrence of steroid estrogens, endocrine-disrupting phenols, and acid pharmaceutical residues in urban riverine water of the Pearl River Delta, South China. AB - A scoping study was conducted to investigate the residues of nineteen pharmaceuticals and personal care products (PPCPs), including 4 natural and 3 synthetic steroid estrogens, 7 endocrine-disrupting phenols, and 5 acid pharmaceuticals in three urban streams and the Major Pearl River at Guangzhou, a megapolis in the Pearl River Delta, South China. Estrone was detected in >60% water samples with a maximum concentration of 65 ng L(-1). Endocrine-disrupting phenols (nonylphenol, bisphenol A, triclosan, 2-phenylphenol, methyparaben, and propylparaben) were found to be widely present at rather high concentrations in the urban riverine water of Guangzhou. Salicylic acid, clofibric acid and ibuprofen were detected in most water samples with maximum concentrations of 2098, 248 and 1417 ng L(-1) respectively, whereas naproxen was less frequently detected and also at lower concentration. Both the detection frequencies and median concentrations of the PPCPs appeared higher during the low-flow season than during the high-flow season. The seasonal difference in PPCPs occurrence was probably attributed to the dilution effect caused by the rainfall. PPCPs in the urban riverine water of Guangzhou originated mainly from random discharge and/or leakage of municipal wastewater. PPCPs contamination in the Major Pearl River may be of a potential environmental issue, especially during the low-flow season. PMID- 18407321 TI - The relationship between maintenance dosages of three vitamin K antagonists: acenocoumarol, warfarin and phenprocoumon. AB - INTRODUCTION: Vitamin K antagonists of the coumarin type are widely used oral anticoagulants. OBJECTIVE: We developed a transition algorithm for the maintenance dosages of three frequently used coumarins: warfarin, phenprocoumon and acenocoumarol. METHODS: The study was conducted at the Leiden Anticoagulation Clinic. Patients were participants in a trial of which the main objective was to compare the quality of an oral anticoagulant therapy with phenprocoumon to warfarin. We included patients who initiated oral anticoagulant therapy and patients who were already using acenocoumarol. Patients were randomized to a treatment with warfarin or phenprocoumon. Patients who were randomized to warfarin switched to phenprocoumon at the end of follow-up. We analysed the switch from acenocoumarol to warfarin or phenprocoumon at the start of follow-up and the switch of warfarin to phenprocoumon at the end of follow-up and calculated the transition factors for stable anticoagulation between these three vitamin K antagonists. RESULTS: Fifty-eight patients switched from warfarin to phenprocoumon, 39 from acenocoumarol to phenprocoumon and 44 from acenocoumarol to warfarin. The maintenance dose of warfarin was 0.41 (95%CI 0.39-0.43) times the maintenance dose of phenprocoumon. The transition factor between acenocoumarol and phenprocoumon was 0.84 (95%CI 0.79-0.89) and between acenocoumarol and warfarin 1.85 (95%CI 1.78-1.92). CONCLUSIONS: We determined the transition factors between warfarin, phenprocoumon and acenocoumarol. With these transition factors physicians are able to estimate the maintenance dose when it is necessary for a patient to switch from one coumarin to the other. PMID- 18407322 TI - Epstein-Barr virus-associated venous thromboembolism: a case report and review of the literature. PMID- 18407323 TI - Cardiac mechano-electric feedback and electrical restitution in humans. AB - Electrical restitution in the heart is the property whereby the action potential duration and conduction velocity of a beat of altered cycle length vary according to its immediacy to the preceding basic beat--the coupling interval, usually the diastolic interval. In general, action potential duration (APD) increases with increasing coupling interval, and the relation between action potential duration and the preceding diastolic interval describes the APD restitution curve. The latter has recently been the focus of considerable interest since the steepness of the initial part of the restitution curve plays an important role in electrical stability and arrhythmogenesis. Mechanical stretch has been shown to alter APD and hence refractoriness either through stretch activated channels or by influencing calcium cycling. Such an effect on refractoriness has been proposed as a mechanism of arrhythmogenesis particularly if spatially inhomogeneities manifest within the heart. Here, we review (1) the spatial and temporal characteristics of APD restitution in humans; (2) previously reported work showing that mechanical loading differentially effects APD of interpolated beats of altered cycle length, and hence alters the slope of the APD restitution curve; and (3) evidence that inhomogeneity of APD restitution slope may be an important factor in arrhythmogenesis. PMID- 18407324 TI - Change of ureteral submucosal anti-reflux implantation into an intussuscepted ileal nipple valve as reflux protection in ureterosigmoidostomy. AB - OBJECTIVES: After submucosal implantation of the ureters in ureterosigmoidostomy failed for different reasons, urologists are confronted with the task of creating a new form of urinary diversion. Therefore, the implantation site of the ureters must be removed because of the risk of secondary malignancy with the consequences of a large colonic defect. TECHNICAL CONSIDERATIONS: Here we describe a technique for reimplantation of both ureters with an intussuscepted ileal nipple in ureterosigmoidostomy with three advantages for the patient in situations in which classical submucosal implantation failed after several years. We were able to avoid conversion into another form of incontinent or continent urinary diversion. To prevent the development of colonic tumors at the implantation site where urothelial meets colonic epithelium, we interposed an ileal segment which avoids this epithelial contact. We were also able to cover a large colonic defect which was mandatory to receive a negative surgical margin when the tumor was resected. CONCLUSIONS: This technique of using an ileal nipple as reflux protection in ureterosigmoidostomy allows the patient to keep the form of urinary diversion he is used to live with, for many decades. PMID- 18407325 TI - Preliminary results of radiation therapy for prostate cancer in human immunodeficiency virus-positive patients. AB - OBJECTIVES: The purpose of this study was to report on the clinical outcomes of patients treated at our institution for prostate cancer (PCa) who had been previously diagnosed as Human immunodeficiency virus (HIV) positive. METHODS: The authors conducted a retrospective study of 14 PCa/HIV patients who were being treated for PCa with external beam radiotherapy, brachytherapy, or a combination of the two. Each patient's prostate-specific antigen (PSA) level, CD4 count, and viral load were obtained before the initial radiation treatment and at the time of their most recent follow-up. In addition, 13 of 14 patients completed a quality of life survey with a social worker on staff and were examined by their radiation oncologist to assess the complications after treatment. RESULTS: Comparing pretreatment data with the data at last follow-up, only 1 patient's PSA level remained above 1.1 ng/mL. The average CD4 count remained stable, increasing from 523 to 577 cells/mm(3), with the lowest final count at 200 cells/mm(3). Viral load increased in only 2 of 14 patients. There were no unusual rectal, urinary, or sexual complications, and no infections related to treatment. CONCLUSIONS: Based on changes in viral load and CD4 count, radiotherapy does not appear to have a long-term negative effect on the immune system. Treatment complications are consistent with HIV-negative patients, giving no evidence that the subset of PCa/HIV patients should be treated differently from PCa patients without HIV when considering radiation therapy. More investigation will be necessary before reliable mortality and morbidity data can be assessed. PMID- 18407326 TI - Testosterone surge: rationale for gonadotropin-releasing hormone blockers? AB - Gonadotropin-releasing hormone (GnRH) agonists currently form the mainstay of management of advanced prostate cancer. They effectively suppress serum testosterone levels, which in turn inhibits tumor growth. However, the initial response to GnRH agonists is a transient increase in the serum testosterone levels. Known as a testosterone surge, this can lead to a worsening of symptoms and can adversely affect survival. Therefore, much interest exists in the development of a new class of drugs-GnRH antagonists-which produce immediate suppression of luteinizing hormone and testosterone without a testosterone surge. The most promising GnRH antagonist to date is degarelix. PMID- 18407327 TI - Massive upper gastrointestinal bleeding from pure metastatic choriocarcinoma in patient with mixed germ cell tumor with subclinical intestinal metastasis. AB - Although testicular germ cell tumors have become curable neoplasms, a better understanding of the clinicopathologic features is needed for the rare manifestations associated with treatment failure. We report a rare case of metastatic pure choriocarcinoma involving the small intestine arising from a testicular mixed germ cell tumor. In a patient who developed massive upper gastrointestinal hemorrhage during treatment, the intestinal metastases and focus of bleeding could only be determined by laparotomy. We propose an approach for the determination of subclinical intestinal metastases of testicular germ cell tumor; the case is discussed in light of similar reports in literature. PMID- 18407328 TI - Incidental prostate cancer detected by cystoprostatectomy in Korean men. AB - OBJECTIVES: To determine the incidence and characteristics of incidental prostate cancer diagnosed by cystoprostatectomy (CPT) in Korean men. METHODS: Thirty-six consecutive male patients scheduled to undergo CPT were prospectively enrolled. The CPT specimens were examined and the clinicopathologic characteristics of incidental cancers compared with those of T1c prostate cancers that had undergone radical prostatectomy. Complete transverse sections of the prostate were taken from the apex to the base at 4-mm intervals. RESULTS: Of the 36 CPT patients, 18 (50%) had incidental prostate cancer. Most of the incidental tumors were confined to the prostate gland, except in 1 patient. Tumor involvement at the prostate apex was found in 3 patients (16.7%), and Gleason scores in 3 cases were 7 to 10. Median tumor volume was 0.08 cm(3) (range, 0.01 to 20.51 cm(3)), and a tumor volume of more than 0.5 cm(3) was identified in 5 patients. Of these incidental prostate cancers, 38.9% (19.4% of all CPT patients) were clinically significant. As compared with the 38 T1c prostate cancer patients, incidental prostate cancer patients were older, had a lower prostate-specific antigen level, a lower grade, smaller tumor volume, and were less likely to have multiple tumors. However, no significant differences were observed between these two groups with respect to apical tumor involvement or tumor confinement to the prostate (P >0.05 for each). CONCLUSIONS: Incidental prostate cancers were diagnosed in 50% of CPT specimens, and 19.4% of these were clinically significant. PMID- 18407329 TI - Intravesical botulinum toxin A injections plus hydrodistension can reduce nerve growth factor production and control bladder pain in interstitial cystitis: a molecular mechanism. PMID- 18407330 TI - Pulse granuloma of urinary bladder associated with interstitial cystitis. AB - Pulse granulomas are uncommon, benign foreign body inflammatory reactions that typically occur in the oral cavity. They are exceedingly rare elsewhere. Here we describe a 35-year-old woman who presented with interstitial cystitis and was found to have an incidental bladder mass. Histological examination of the biopsy revealed a pulse granuloma. The salient histopathologic features include corrugated hyaline rings and amorphous hyaline conglomerations within the connective tissue stroma of the lamina propria, admixed with chronic inflammatory cells with multinucleated foreign-body type giant cells. To our knowledge this is the first reported case of a pulse granuloma in the urinary bladder. PMID- 18407331 TI - Treatment of occult reflux lowers the incidence rate of pediatric febrile urinary tract infection. AB - OBJECTIVES: To examine whether vesicourethral reflux diagnosed by positioned instillation of contrast (PIC-VUR) shows clinical importance by comparing the incidence rates of febrile urinary tract infection (FUTI) before and after treatment of PIC-VUR. METHODS: Beginning in 2001 we used a multi-institutional registry to prospectively enroll consecutive pediatric patients with a history of FUTI without VUR according to voiding cystourethrogram (VCUG) and yet who show PIC-VUR. Treatment of PIC-VUR was with prophylactic antimicrobials or antireflux surgery. The post-treatment occurrence of FUTI was tracked. RESULTS: A total of 14 centers enrolled 118 patients (mean age, 7.2 years; range, 0.5 to 20 years). Parents self-selected the treatment of PIC-VUR as endoscopic injection (104), ureteral reimplantation (3), or antimicrobial prophylaxis (11). Study intervals surveying for FUTI before PIC (mean, 12 months; range, 1 to 17 years) and after PIC treatment (mean, 11 months; range, 0 to 3 years) were not significantly different. Overall the incidence rate for FUTI decreased significantly from 0.16 per patient per month before PIC-VUR treatment to 0.008 per patient per month after treatment (rate ratio 20; 95% confidence interval 11 to 36). The post treatment rate of FUTI in patients treated with antibiotics versus surgery was not significantly different (rate ratio 2.5; 95% confidence interval 0.33 to 27). CONCLUSIONS: The diagnosis of PIC-VUR is clinically important because children treated for PIC-VUR with either antimicrobial prophylaxis or surgery show a significant reduction in the incidence rate of FUTI. This is the basis for a current prospective study randomizing patients with PIC-VUR to treatment or observation. PMID- 18407332 TI - Comparative experimental evaluation of guidewire use in urology. AB - OBJECTIVES: To evaluate the characteristics and mechanical properties of five different guidewires principally used in urology to provide a guide to the best choice in every situation. This objective was achieved by performing mechanical tests on each guidewire and on the same guidewire in relationship to a real human ureter. Finally, the mechanical properties of a real human ureter were evaluated experimentally. METHODS: Five different guidewires with a 0.035-in. external diameter were studied to determine their mechanical properties of interest: tip buckling, shaft permanent deformation, guide friction on the human ureter, and the force at which the tip perforated a real human ureter. RESULTS: Nitinol guidewires with hydrophilic coating had the least dangerous tip in the tip buckling and tip piercing force tests and the lowest friction values on the friction guide test. The shaft of the polytetrafluoroethylene-coated guidewires was stiffer on the bending resistance and plastic deformation tests. CONCLUSIONS: The ideal endourologic guidewire should be sufficiently flexible and lubricated to negotiate a tortuous ureter and pass possible obstructions. However, high strength against bending is also required when we need to pass a stent or catheter over a guidewire. PMID- 18407333 TI - Veterans Affairs Cooperative Studies Program study 553: Chemotherapy after prostatectomy, a phase III randomized study of prostatectomy versus prostatectomy with adjuvant docetaxel for patients with high-risk, localized prostate cancer. PMID- 18407334 TI - A new cause of male infertility after cisplatin exposure: the effect of cisplatin on Y chromosomes. AB - OBJECTIVES: To investigate the impact of cisplatin (CP) on the testes-specific protein, Y-linked (TSPY) gene situated on the Y chromosome. METHODS: The control group consisted of 10 rats. Group IIA consisted of 15 rats that underwent orchiectomy and received three cycles of 1 mg/kg, 2.5 mg/kg, or 5 mg/kg CP. Group IIB was exposed to the same doses of three cycles of chemotherapy but was examined after 3 months of chemotherapy. Group III was exposed to the same doses of chemotherapy without initial orchiectomy. Reverse transcriptase polymerase chain reaction for TSPY messenger ribonucleic acid (mRNA) and immunohistochemical staining for histone 2B were performed on the testes. Results were evaluated by one-way analysis of variance. RESULTS: Compared with the controls, the expression of TSPY mRNA in Group IIA after exposure to 1 mg/kg CP did not change; however, mRNA levels after exposure to 2.5 mg/kg and 5 mg/kg CP were decreased by 40% and 78%, respectively. In Group III after exposure to the same doses of CP, mRNA levels decreased by 30%, 87.5%, and 88%, respectively. The expression of TSPY was at normal levels except in rats that received 5 mg/kg CP in Group IIB. Immunohistochemical study revealed that histone 2B expression was decreased in a dose-dependent manner. None of the rats from any of the groups died during the study period. CONCLUSIONS: Decreased TSPY expression after CP exposure might be another mechanism for male infertility. PMID- 18407335 TI - Is there evidence for a role of Propionibacterium acnes in prostatic disease? PMID- 18407336 TI - Large renal cavernous hemangioma with renal vein thrombosis: case report and review of literature. AB - Kidney cavernous hemangiomas are rare. Renal cavernous hemangioma complicated by secondary chronic thrombosis of the renal vein has not been previously reported. A 17-year-old boy was initially diagnosed with a renal mass with tumor thrombosis. Radical nephrectomy was performed, and the pathologic examination revealed a renal cavernous hemangioma with chronic renal vein thrombosis. We also reviewed the previous diagnostic and treatment methods described in previous reports. PMID- 18407337 TI - Urinary quality of life after prostatectomy or radiation for localized prostate cancer: a prospective longitudinal cross-cultural study between Japanese and U.S. men. AB - OBJECTIVES: To compare the evolution of urinary dysfunction and associated distress (bother) during the first 2 years after radical prostatectomy (RP) or external beam radiation therapy (EBRT) between Japanese and U.S. men with localized prostate cancer. METHODS: A total of 477 Japanese men and 385 U.S. men with localized prostate cancer who underwent RP or EBRT participated in paired longitudinal outcomes studies. We evaluated urinary control and distress using the University of California-Los Angeles, Prostate Cancer Index and urinary irritation/obstruction using the American Urological Association Symptom Index (AUASI) before and 1, 2 to 3, 4 to 6, 8, 12, 18, and 24 months after treatment. We used general linear mixed modeling adjusting for subject characteristics to assess changes in these domains. RESULTS: Multivariate analyses revealed a nonlinear trend of recovery and an interaction between this trend and country with regard to urinary function (control) and bother after RP (both P <0.0001). Among the men who received EBRT, the recovery trend of AUASI and urinary bother also differed significantly by country (both P <0.0001). CONCLUSIONS: Japanese and U.S. men differed in their patterns of urinary recovery up to 24 months after curative therapy for localized prostate cancer. PMID- 18407338 TI - Serum inhibin--not a cause of low testosterone levels in hypogonadal prostate cancer? AB - OBJECTIVES: High-grade prostate cancer is associated with low serum testosterone levels, which generally recover after radical prostatectomy. The cause of this low testosterone level is unclear, and it has been hypothesized that cancer cells produce a factor that disturbs the pituitary-gonadal axis. Inhibin is a hormone that has a negative feedback effect on this axis. The aim of this study was to investigate the role of serum inhibin in patients with prostate cancer. METHODS: The serum hormone levels of the pituitary-gonadal axis, including inhibin levels, in patients with prostate cancer were compared with those in patients with benign prostatic hyperplasia. Testosterone levels of less than 3 ng/mL were classified as hypogonadal. Prostate cancer was classified according to Gleason score as high grade (Gleason score 7 to 10) or low grade (Gleason score 2 to 6). RESULTS: A total of 196 men (126 with prostate cancer and 70 with benign prostatic hyperplasia) were entered into the study. The serum inhibin levels did not differ significantly between the patients with benign prostatic hyperplasia and those with prostate cancer (150.0 versus 131.75 pg/mL, P = 0.062), between men with hypogonadal and eugonadal disease (143.0 versus 146.5 pg/mL, P = 0.573), or between those with low-grade and high-grade cancer (151.5 versus 146.0 pg/mL, P = 0.830). Men with high-grade cancer had lower levels of serum testosterone than did those with low-grade cancer (3.49 versus 4.09 ng/mL, P = 0.056). CONCLUSIONS: The results of our study have shown that although high-grade prostate cancer is associated with low serum testosterone levels, inhibin does not appear to be the cause of this phenomenon. PMID- 18407339 TI - New aspects of vulvar cancer: changes in localization and age of onset. AB - OBJECTIVE: To characterize the changes in incidence, age of disease onset, tumor site and patients characteristics in women with invasive vulvar cancer in a German University Hospital unit over a 28-year period. METHODS: The clinical records for women treated for invasive vulvar cancer from 01/1980 until 06/2007 were analyzed. We performed a retrospective analysis for three 9-year periods: 1/1980 to 02/1989; 3/1989 to 04/1998 and 05/1998 to 06/2007. For each cohort, the number of cases treated, age of disease onset, tumor site and further characteristics were extracted and statistically evaluated. RESULTS: A total of 224 patients with vulvar cancer were identified between 1/1980 and 6/2007. The number and mean age changed significantly over time: between 1/1980 and 02/1989 53 women with a mean age of 65.6 years were treated for invasive vulvar cancer, between 03/1989 and 04/1998 this number increased to 69 women with a mean age of 63.9 years and in the last period, 102 women with a mean age of 57.0 years were treated for vulvar cancer. The total increase was 192%. In the first period 11% of the women were aged 50 years or less compared with over 41% in the third period (p=0.001). Two-third of the tumors women aged<50 years were HPV-positive. Significant changes in the tumor site were observed; from labial position to the region between clitoris and urethra: 37% in the last period compared with 19% in the first period (p>0.05). CONCLUSIONS: Although in the literature the incidence of invasive cancer has been reported to be stable or only minimally increased, the results of this study show that the number of patients presenting with invasive vulvar cancer has doubled within the last three decades at one university hospital unit in Germany, with a nearly 4-time increase in younger patients (+372%) due to HPV high risk infection. The tumor localization changed significantly from the labia to the area between the clitoris and urethra. Assuming that these limited data reflect the general trend in the incidence of HPV-induced vulvar cancer, widely-implemented prophylactic quadrivalent HPV vaccination, which has been proven to be highly effective against anogenital disease, could make an important contribution to the reduction of the risk of vulvar carcinomas in younger women. PMID- 18407340 TI - Bilateral oophorectomy before 50 years of age is significantly associated with the metabolic syndrome and Framingham risk score: a controlled, population-based study (HUNT-2). AB - OBJECTIVE: Bilateral oophorectomy (BOE) is often recommended in order to prevent cancer at hysterectomy for non-malignant diseases and when familial risk of ovarian and breast cancer has been identified. Surgical menopause increases the risk of cardiovascular mortality, however, the intervening mechanisms are not clear. We compared the prevalence of the metabolic syndrome (MetS) and Framingham cardiovascular risk scores in women with BOE before 50 years of age to age matched controls in a population-based study. METHODS: 20,765 women aged 40-69 years were invited to a health study (HUNT-2 Norway 1995-97) and 17,650 (85%) attended. We compared 263 women with BOE before 50 years of age [63 with intact uterus (BO1 group), and 200 with hysterectomy also (BO2 group)] with 3 age matched controls per case (n=789). Data on demographic, somatic, mental, and lifestyle variables, physical measurements and blood tests were obtained. RESULTS: The BO1 and BO2 groups did not differ significantly regarding risk variables, and 4% had natural menopause. The combined BOE group had increased prevalence of MetS compared to controls according to the International Diabetes Federation's definition (47% versus 36%; p=.001) and the revised NCEP ATP III definition (35% versus 25%; p=.002), which remained after adjustments (for reproductive, global health, and lifestyle variables). The prevalence of Framingham risk score > or =10% was higher in cases (22%) versus controls (15%) p=.005. CONCLUSION: The higher prevalence of MetS and increased Framingham risk scores in women with bilateral oophorectomy before 50 years of age suggests that these women may be at higher risk of type 2 diabetes and cardiovascular disease compared to their counterparts in the general population. PMID- 18407341 TI - Carboplatin dosing in obese women with ovarian cancer: a Gynecologic Oncology Group study. AB - BACKGROUND: Carboplatin dosing for gynecologic malignancies is traditionally based on the Jelliffe formula that lacks dose adjustment for weight. Obese women may therefore receive a sub-therapeutic carboplatin dose. This study assessed the association between BMI and outcome for ovarian cancer patients treated with carboplatin-based chemotherapy. METHODS: An analysis of patients treated with carboplatin and paclitaxel on Gynecologic Oncology Group (GOG) protocol 158 was performed. The dose of carboplatin for each patient was based on an area under the curve of 7.5 and a glomerular filtration rate (GFR) derived from the Jelliffe formula which is derived from age and serum creatinine. Patients were stratified based on body mass index (BMI). RESULTS: A total of 387 patients were included in the analysis. The patients were stratified into three groups: normal weight (BMI<25.0, 50%), overweight (BMI 25-29.9, 32%) and obese (BMI > or = 30.0, 18%). Compared to pretreatment values, the obese patients had a lower relative decrease in their platelet counts (-25% for BMI > or = 30 vs. -61% for BMI<25) (p = 0.01). Similar trends were noted for relative changes in hemoglobin (p = 0.006) and hematocrit (p = 0.002). Dose reductions were required in 34% of normal weight compared to 21% of the obese women (p = 0.004). There was a trend toward increased risk for disease progression in women with a BMI > or = 30 (RR: 1.25, 95% CI: 0.93-1.69, p = 0.14). CONCLUSION: Obese ovarian cancer patients treated with carboplatin experience substantially less toxicity than normal weight women. The lower toxicity suggests that obese patients may be receiving a substandard drug dose. PMID- 18407343 TI - Validity of self-reported mammography in a multicultural population in Israel. AB - OBJECTIVE: To validate self-reported mammography against claims records in women aged 52-74 living in Israel and belonging to the Jewish (non-orthodox pre-1989 native or former Soviet Union immigrant or ultra-orthodox) or Arab populations. METHODS: In a spring 2007 random telephone survey, 1550 women receiving healthcare at Maccabi Health Services were asked whether they had had a mammography during the previous 2 years. The same information was obtained from claims records and treated as the gold standard. RESULTS: Self-reported mammography and claims records disagreed for 17.4%. Compared to the other populations, Arab women tended to report more often that they had obtained a mammogram when it was not registered in the claims data (specificity=47.3%, 95% CI%=38.4, 56.3). Ultra-orthodox women more often failed to report having had the mammogram while the claims records indicated they had had (sensitivity=90.3%, 95% CI%=86.1, 93.6). CONCLUSION: Agreement between self-reported mammography and claims records depends on cultural and socioeconomic factors. PMID- 18407342 TI - Role of systemic endocannabinoid CB-1 receptor antagonism in the acquisition and expression of fructose-conditioned flavor-flavor preferences in rats. AB - Rats learn to prefer a flavor mixed into a fructose-saccharin solution over a different flavor mixed into a saccharin-only solution which is considered to be a form of flavor-flavor conditioning. Fructose-conditioned flavor preferences are impaired by systemic dopamine D1 and to a lesser degree, D2 receptor antagonism as well as by NMDA, but not opioid, receptor antagonism. Given the emerging role of the endocannabinoid system in mediating hedonically-driven food intake, the present study examined whether systemic administration of the inverse CB-1 receptor agonist, AM-251 would alter fructose-conditioned flavor preferences. In Experiment 1, food-restricted rats were trained over 10 sessions (30 min/day) to drink a fructose-saccharin solution mixed with one flavor (CS+/Fs) and a less preferred saccharin-only solution mixed with another flavor (CS-/s). Subsequent two-bottle tests with the two flavors in saccharin (CS+/s, CS-/s) occurred 15 min following counterbalanced pairs of AM-251 doses of 0, 0.1, 1 or 3 mg/kg. Preference for CS+/s over CS-/s following vehicle treatment (74%) was significantly reduced by the 0.1 (67%) and 1 (65%) AM-251 doses, whereas CS+/s, but not CS-/s intake was significantly reduced by the 1 and 3 mg/kg AM-251 doses. In Experiment 2, rats received systemic injections of AM-251 (1 mg/kg) or vehicle prior to the 10 CS+/Fs and CS-/s training sessions. In subsequent two-bottle tests (drug-free) the AM-251 and control groups displayed similar preferences for the CS+ flavor (66% vs. 69%). Experiment 3 demonstrated that AM-251 significantly decreased chow intake (24 h), and 1-h intakes of fructose-saccharin and saccharin only solutions in ad libitum-fed rats. These data indicate that functional CB-1 receptor antagonism significantly reduces the expression, but not the acquisition of fructose-conditioned flavor-flavor preferences. The endogenous endocannabinoid system is therefore implicated in the maintenance of this form of learned flavor preferences. PMID- 18407344 TI - Non-invasive ultrasonographic characterization of phenotypic changes during embryo development in non-anesthetized mice of different genotypes. AB - This study aimed to determine phenotypic changes during embryo development in the mouse, and the influence of genetic background, by non-invasive real-time ultrasonography. Serial scannings were performed from Day 4.5 after the appearance of the vaginal plug in a total of 34 adult mice of different strains (9BALB/c, 10C57BL/6 and 15 CD1). Embryonic vesicle diameter was measurable from Day 4.5 of pregnancy. Images of embryos were obtainable from Day 5.5 of gestation onwards, and crown-rump length and trunk parameters (diameter and area) were measured. At more advanced stages, the positions of fetuses prevented accurate measurement of crown-rump length; therefore, head diameters (occipito-snout length and biparietal diameter) were used as an alternative index of fetal size. All measurements correlated highly with gestational age (P<0.0005). No significant differences were observed between genotypes in early pregnancy, but during the last week of gestation trunk measurements were larger in CD1 embryos (P<0.05) while head diameters were larger in C57BL/6 conceptuses (P<0.05). There was a significant effect of genetic background on heart rate throughout pregnancy; although heart rate was similar in CD1 and C57BL/6 embryos (154.5+/ 2.8 and 147.8+/-4.5 beats/min, respectively), it was significantly lower in BALB/c mice (127.0+/-2.1; P<0.005 vs. CD1 and C57BL/6). PMID- 18407345 TI - Doppler evaluation of maternal and foetal vessels during normal gestation in queen. AB - The aim of this work is to evaluate the haemodynamic characteristics of maternal and foetal vessels during normal pregnancy in queens, using colour Doppler and pulsed wave Doppler ultrasonography, in order to obtain information about maternal and foetal circulation. The blood waveforms of the uteroplacental arteries, aorta, caudal cava vein and umbilical cord of the fetuses were recorded weekly in seven healthy pregnant queens. Also, the measurements of peak systolic, end diastolic velocities, resistance and pulsatility indices were carried out. Uteroplacental blood flow was biphasic while the ones of the umbilical artery and aorta were first systolic and then diastolic. The caudal cava vein showed a typical waveform of venous vessels. During gestation the EDV and PSV of foetal vessels increased (alpha<0.05) while the PI and RI of all vessels examined decreased (alpha<0.05) except for the IP of the aorta. The Doppler ultrasonography, also in queens, can be used to evaluate the characteristics of maternal and foetal vessel flow and their progressive changes during pregnancy. This study can be considered the basis for further contribution in diagnosing and monitoring high-risk pregnancies in Veterinary Medicine. PMID- 18407346 TI - Effects of a self-etching resin coating system to prevent demineralization of root surfaces. AB - OBJECTIVES: The purpose of this study was to evaluate the ability of an experimental self-etching resin coating system to protect against demineralization of the root surface in vitro, comparing its effectiveness with two commercial one-step self-etching adhesives. METHODS: The experimental resin coating system (EC), consisting of a self-etching primer and an acrylate-based resin, or the two commercial self-etching adhesives, Adper Prompt-L Pop (AD) and Clearfil Tri-S Bond (S3), was applied to the cementum of a human extracted tooth root. The specimens were immersed in acetate buffer at pH 5.5 for 4 weeks, and the degree of demineralization underneath the materials was evaluated by microscopic observation and contact microradiography. Each material's sealing and interfacial ultrastructure on the cementum was morphologically examined by scanning electron microscopy, and their bonding ability to cementum was determined by microtensile bond strength (MTBS) tests. RESULTS: Application of AD and S3 showed limited effects to protect against demineralization, but no demineralization was observed for EC. EC showed greater coating thickness than the other two materials, with formation of integral hybridized layers with cementum. Although the MTBS values for EC were significantly lower than those for AD and S3, cohesive failure in resins was the prevalent fracture mode for EC, showing no possible disadvantages in bonding ability. SIGNIFICANCE: The experimental resin coating system effectively prevented acid demineralization on root surfaces due to its ability to produce an appropriately thick coating and an integral hybridized layer. PMID- 18407347 TI - A weight-of-evidence approach to assessing the ecological impact of organotin pollution in Dutch marine and brackish waters; combining risk prognosis and field monitoring using common periwinkles (Littorina littorea). AB - In the present study an integrated ecological risk assessment based on multiple lines of evidence (LOEs) was evaluated in order to better assess the risk from TBT in Dutch harbours and open coastal waters. On the basis of spatial distributions of measured tributyltin (TBT) concentrations in sediments and suspended matter, predictions of the intersex index (ISI) in Littorina littorea and the ecological risk expressed as the Potentially Affected Fraction (PAF) of species were made. The results were compared to actual ISI measurements and presence of L. littorea in the field. The PAF calculated on the basis of TBT levels for open coastal waters ranged from 4.2% to 15.3%; for harbours it ranged from 3.5% to 26.9%. Significant intersex levels were observed only in waters where the risk was calculated above 10% PAF. This study suggests that the absence of L. littorea from some harbours with high ecological risk values can be explained by high TBT concentrations. A call is made for the use of integrated approaches like weight-of-evidence (WOE) to help practitioners improve ecological risk assessment. PMID- 18407348 TI - The effect of pretreating morselized allograft bone with rhBMP-2 and/or pamidronate on the fixation of porous Ti and HA-coated implants. AB - BMPs stimulate new bone formation, but may also accelerate bone resorption. We added rhBMP-2 and pamidronate to morselized bone allograft packed around uncemented HA-coated and non-coated porous Ti implants in sixteen dogs. Each dog received four implants where the allograft was added (1) nothing, (2) BMP, (3) BP, and (4) BMP+BP. After four weeks, the untreated control implants had better mechanical fixation than all other treatment groups. The rhBMP-2-treated group had abundant formation of new bone on and around the implant. However, almost all allografts were resorbed, rendering the implant mechanically unstable. In the pamidronate-treated group the allograft was preserved, but the implants were covered by fibrous tissue and there was almost no new bone formation. This was also the case for the combined BMP+BP group, although fibrous tissue was absent on these implants. The HA-coated implants had more than twice as good mechanical fixation and improved osseointegration compared to the corresponding Ti implants. RhBMP-2 raised the total metabolic turnover of bone within the allograft with a net negative result on implant fixation. Pamidronate virtually blocked bone metabolism, also when combined with rhBMP-2. The results warrant a conservative approach and emphasize the importance of identifying a therapeutic window for these substances prior to clinical use. PMID- 18407349 TI - Cyclooxygenase-2 derived PGE2 and PGI2 play an important role via EP2 and PPARdelta receptors in early steps of oil induced decidualization in mice. AB - Differentiation of endometrial stromal cells into decidual cells (decidualization) is prerequisite for blastocyst implantation. Different prostanoids are shown to be involved in the cascade of events found in implantation and decidualization. Previous reports described that cyclooxygenase 2 (COX2) derived prostacyclin (PGI2) plays an important role via peroxisome proliferator activated receptor (PPARdelta) nuclear receptor in implantation and decidualization. Herein, we investigated the role of COX2 derived PGE2 and PGI2 and examined the protein expression and regulation of COX1, COX2, membrane-bound prostaglandin E synthase (mPGES-1), prostaglandin I synthase (PGIS), PGE2 receptor (EP2) and PPARdelta in hormone primed oil infused uterine horn as well as in non-infused uterine horn (control horn). Our results show that selective COX2 inhibitor (Nimesulide) inhibits decidualization while COX1 inhibitor (SC560) does not affect decidualization. COX2, mPGES-1, PGIS, EP2 and PPARdelta immunostaining are strongly observed at 24 h and 48 h in oil-induced horn and than significantly reduced at 72 h and 120 h and absent in non-infused horn. However COX1 immunostaining is observed in infused as well as in non-infused horn. Our immunohistochemical studies corroborated well with follow up western blotting of the same proteins. PGE2 and PGI2 products were also elevated at 24h and 48 h after oil induction in infused horn in comparison to control horn. Our data suggest that COX2 derived both PGE2 and PGI2 mediate its function via EP2 and PPARdelta receptors in early steps of decidualization in mice. PMID- 18407350 TI - Bcl-2 suppresses Ca2+ release through inositol 1,4,5-trisphosphate receptors and inhibits Ca2+ uptake by mitochondria without affecting ER calcium store content. AB - Cell survival is promoted by the oncoprotein Bcl-2. Previous studies have established that one of the pro-survival actions of Bcl-2 is to reduce cellular fluxes of Ca2+ within cells. In particular, Bcl-2 has been demonstrated to inhibit the release of Ca2+ from the endoplasmic reticulum. However, the mechanism by which Bcl-2 causes reduced Ca2+ release is unclear. In the accompanying paper [C.J. Hanson, M.D. Bootman, C.W. Distelhorst, T. Maraldi, H.L. Roderick, The cellular concentration of Bcl-2 determines its pro- or anti apoptotic effect, Cell Calcium (2008)], we described that only stable expression of Bcl-2 allowed it to work in a pro-survival manner whereas transient expression did not. In this study, we have employed HEK-293 cells that stably express Bcl-2, and which are, therefore, protected from pro-apoptotic stimuli, to examine the effect of Bcl-2 on Ca2+ homeostasis and signalling. We observed that Bcl-2 expression decreased the Ca2+ responses of cells induced by application of submaximal agonist concentrations. Whereas, decreasing endogenous Bcl-2 concentration using siRNA potentiated Ca2+ responses. Furthermore, we found that Bcl-2 expression reduced mitochondrial Ca2+ uptake by raising the threshold cytosolic Ca2+ concentration required to activate sequestration. Using a number of different assays, we did not find any evidence for reduction of endoplasmic reticulum luminal Ca2+ in our Bcl-2-expressing cells. Indeed, we observed that Bcl-2 served to preserve the content of the agonist-sensitive Ca2+ pool. Endogenous Bcl-2 was found to interact with inositol 1,4,5-trisphosphate receptors (InsP3Rs) in our cells, and to modify the profile of InsP3R expression. Our data suggest that the presence of Bcl-2 in the proteome of cells has multiple effects on agonist-mediated Ca2+ signals, and can abrogate responses to submaximal levels of stimulation through direct control of InsP3Rs. PMID- 18407351 TI - Bone marrow fibroblasts induce expression of PI3K/NF-kappaB pathway genes and a pro-angiogenic phenotype in CLL cells. AB - Microarray-based gene expression profiling (GEP) was used to study how stroma modulates the survival of CLL cells in an in vitro coculture model employing the murine fibroblast cell line M2-10B4. CLL cells cultured in direct contact with the stromal layer (STR) showed a significantly better survival than cells cultured in transwell (TW) inserts above the M2-10B4 cells. STR as compared to TW conditions induced a significant up-regulation of PI3K/NF-kappaB pro-survival pathway genes and mediated a pro-angiogenetic switch in the CLL cells by up regulation of vascular endothelial growth factor (VEGF) and osteopontin (OPN) and down-regulation of the anti-angiogenetic molecule thrombospondin-1 (TSP-1). PMID- 18407352 TI - Biomonitoring of cyanotoxins in two tropical reservoirs by cladoceran toxicity bioassays. AB - This study evaluates the potential for the use of cladocerans in biomonitoring of cyanobacterial toxins. Two zooplankton species (Daphnia gessneri and Moina micrura) were cultivated in the laboratory for use in acute (48 h) and chronic (10 days) bioassays. Water samples were collected from two reservoirs and diluted in mineral water at four concentrations. Survivorship in the acute bioassays was used to calculate LC50, and survivorship and fecundity in chronic bioassays were used to calculate the intrinsic population growth rate (r) and the EC50. Analysis of phytoplankton in the water samples from one reservoir revealed that cyanobacteria were the dominant group, represented by the genera Anabaena, Cylindrospermopsis, and Microcystis. Results of bioassays showed adverse effects including death, paralysis, and reduced population growth rate, generally proportional to the reservoir water concentration. These effects may be related to the presence of cyanobacteria toxins (microcystins or saxitoxins) in the water. PMID- 18407353 TI - Effects of Aroclor 1254 on oxidative stress in developing Xenopus laevis tadpoles. AB - Over the last decades, amphibians decline has been reported worldwide. Exposure to polychlorinated biphenyls (PCBs) is one of the possible causes in addition to climate changes, UV-radiation or habitat destruction. In the present study, we tested the hypothesis that PCBs could induce oxidative stress in young tadpoles. Developing Xenopus laevis were exposed from 2- to 5-d postfertilization (pf) to 0.1 or 1 mg/l of Aroclor 1254. Lipid peroxidation and antioxidant systems (SOD, CAT, GST, GPx, GR activities and t-GSH level) were investigated in whole organisms. Exposure to both concentrations did not impact on the survival and development whereas the average body weight decreased. Exposure to 1 mg/l of Aroclor 1254 induced a significant (p<0.05) increase of GST activity when compared to controls 0 and DMSO. The other antioxidant enzymes and LPO evaluation remained unchanged. Our results demonstrate that exposure of X. laevis tadpoles to environmental concentrations of Aroclor 1254 interfere with normal growth. They also highlight that very young X. laevis tadpoles express antioxidant systems. PMID- 18407354 TI - Interactions of 2,4,6-trinitrotoluene (TNT) with xenobiotic biotransformation system in European eel Anguilla anguilla (Linnaeus, 1758). AB - The aim of the present study was to investigate the interaction of 2,4,6 trinitrotoluene (TNT) with liver biotransformation enzymes in European eel Anguilla anguilla (Linnaeus, 1758). Eels were exposed to 0.5, 1 and 2.5mg/l nominal concentrations of TNT for 6 and 24h. Modulation of CYP1A1, UDPGT and GST genes was investigated by real-time PCR. Total CYP450 content, NADPH cytochrome c reductase activity, CYP1A and CYP2B-like activities, such as EROD, MROD and BROD, as well as GST and UDPGT activities, were measured by biochemical assays. An in vitro study was performed on EROD in order to evaluate catalytic modulation by TNT. No modulation of the CYP1A1 gene or protein was observed in TNT-exposed eels. On the other hand, a significant decline of EROD and MROD activities was observed in vivo. An increase in NADPH cyt c reductase, and phase II enzymes (UDPGT and GST) were observed at both gene expression and activity levels. The overall results indicated that TNT is a potential competitive inhibitor of CYP1A activities. A TNT metabolic pathway involving NADPH cyt c reductase and phase II enzymes is also suggested. PMID- 18407355 TI - A randomized trial of 2% chlorhexidine tincture compared with 10% aqueous povidone-iodine for venipuncture site disinfection: Effects on blood culture contamination rates. AB - BACKGROUND: Contaminated blood cultures have been recognized as a bothersome issue, and continue to cause frustration for clinicians. Skin antiseptics can prevent blood culture contamination. To our knowledge, there have been no randomized studies to compare 2% alcoholic chlorhexidine and 10% aqueous povidone iodine for venipuncture site disinfection. OBJECTIVE: This study aimed to evaluate the efficacy of venipuncture site disinfection with 2% chlorhexidine in 70% alcohol and 10% aqueous povidone-iodine in preventing blood culture contamination. PATIENTS AND METHODS: A prospectively randomized investigator blinded trial was conducted in all patients in the medical wards and emergency room (ER) at King Chulalongkorn Memorial Hospital, Bangkok, Thailand, from August to October, 2006. Venipuncture sites were disinfected with 2% chlorhexidine in 70% alcohol or 10% aqueous povidone-iodine, and blood cultures were taken by students, residents, or nurses. RESULTS: Of 2146 blood cultures, 108 (5.03%) were contaminated with skin flora. The blood culture contamination rate with 2% alcoholic chlorhexidine was 3.2% (34 of 1068), compared with a rate of 6.9% (74 of 1078) (P<0.001) with 10% aqueous povidone-iodine. In medical wards, the contamination rates were 2.6% and 3.9% with 2% alcoholic chlorhexidine and 10% aqueous povidone-iodine (P=0.2). In ER, the contamination rates were 4.3% and 12.5% with 2% alcoholic chlorhexidine and 10% aqueous povidone-iodine (P<0.001). The most common contaminant was coagulase-negative Staphylococcus (80.6%), followed by Corynebacterium (7.4%), Micrococcus (6.5%), and Bacillus (5.5%). CONCLUSION: Two percent alcoholic chlorhexidine is superior to 10% aqueous povidone-iodine for venipuncture site disinfection before obtaining blood cultures. PMID- 18407356 TI - 12th International Symposium Of Chemistry, Miercurea Ciuc (Csikszereda), Romania, 5th to 8th of October, 2006. 7th International Symposium and Summer School on Bioanalysis, Pecs, Hungary, 10th to 17th of June, 2007. PMID- 18407357 TI - Relationship of varying patterns of cytokine production to the anorexic and neuroendocrine effects of repeated Staphylococcal enterotoxin A exposure. AB - Staphylococcal enterotoxin A (SEA) is a superantigen that stimulates T cells and induces the production of multiple cytokines. Previous studies have shown that SEA augments gustatory neophobia and activates the hypothalamic-pituitary-adrenal (HPA) axis. This study aimed to determine if the cytokine response, behavioral effects, and HPA axis activation persisted after repeated SEA treatment. Male C57BL/6J mice were given 1-4 intraperitoneal injections of 5 microg SEA, after which food intake, corticosterone, or peripheral cytokines were measured. In a series of experiments, it was found that secondary exposure to SEA two or three days after priming increased corticosterone, but attenuated splenic TNFalpha, while augmenting IL-1beta, IL-2, and IFNgamma. The anorexic response was intact after secondary exposure, but absent after a third injection, which was still able to elevate corticosterone. It is unlikely that IL-1 mediated the persistent effects on corticosterone, since this was increased in groups lacking corticosterone elevations. Similarly, TNFalpha was only modestly elevated under repeated SEA conditions that elevated plasma corticosterone. This attenuation appeared to be inversely related to the levels of IL-10, the production of which incrementally rose with each successive injection. In conclusion, repeated exposure to SEA activates the HPA axis and alters behavior. However, there may be dissociation between the behavioral and endocrine effects of SEA with increased SEA exposure. Furthermore, it is possible that while TNFalpha was previously shown to be important in response to acute SEA-induced HPA axis activation, further exposure to SEA elicits other cytokines that may exert neuromodulatory effects through sensitization and/or synergistic mechanisms. PMID- 18407358 TI - Novel autoantigens recognized by CSF IgG from Hashimoto's encephalitis revealed by a proteomic approach. AB - To identify the target of IgG autoimmune response in Hashimoto's encephalopathy (HE), we studied the binding of IgG present in serum and cerebro-spinal fluid (CSF) from six patients with HE and 15 controls to human central nervous system (CNS) white matter antigens by 2D-PAGE and immunoblotting and by immunohistochemistry. We found that CSF IgG from HE patients specifically recognized 3 spots, which were identified as dimethylargininase-I (DDAHI) and aldehyde reductase-I (AKRIAI). DDAHI was present in two isoforms recognized respectively by five and four HE patients; immunohistochemistry with anti-DDAHI antiserum depicted endothelial cells in normal human CNS. AKRIAI was recognized by three HE CSF and this enzyme was widely distributed on neurons and endothelia by immunohistochemistry. IgG from HE CSF immunostained both neuronal and endothelial cells in mouse CNS. The presence of these autoantibodies selectively in the CSF of HE patients may have important diagnostic and pathogenetic implications, since the autoimmune response to these enzymes may lead to vascular and/or neuronal damage, two major mechanisms involved in the pathogenesis of HE. PMID- 18407359 TI - Novel pharmacological strategies for driving inflammatory cell apoptosis and enhancing the resolution of inflammation. AB - Resolution of inflammation requires the effective downregulation of key inflammatory cells such as neutrophils and eosinophils, which normally undergo programmed cell death (apoptosis) to enable their detection and removal by phagocytes such as macrophages. Dysregulation of this process is thought to contribute to the pathogenesis and progression of chronic inflammatory disorders such as chronic obstructive pulmonary disease, asthma, rheumatoid arthritis, allergic rhinitis and inflammatory bowel disease. Importantly, knowledge of the signalling pathways responsible for the induction and execution of granulocyte apoptosis and the phagocytic removal of apoptotic cells continues to increase and, with it, the potential for incisive pharmacological intervention. In this article, we highlight pharmacological strategies that could be used to drive the resolution of inflammation by augmenting apoptosis of inflammatory cells. PMID- 18407360 TI - Role of fosB in behaviours related to morphine reward and spatial memory. AB - The immediate early genes (IEGs) have been suggested to be implicated in mechanisms of addiction, as well as in learning and memory processes. fosB, which belongs to IEG, has been reported to have pleiotropic impact on response to psychoactive drugs, as well as motivational and stress-related behaviours. In the present study, we used mice with constitutive knock-out of fosB in order to study fosB role in mouse phenotype. We studied rewarding properties of morphine (10mg/kg i.p.) in conditioned place preference (CPP) paradigm. Additionally, we studied fosB role in spatial memory and spatial working memory using elevated plus maze model of spatial learning (EPMSL) and delayed non-match to place task (DNMTP). In further studies, locomotor, depressive-like and anxiety-like behaviours were measured. Rewarding effects of morphine in fosB -/- mice were abolished whereas spatial learning was impaired. On the other hand, we found no significant differences in locomotor activity, depression-like and anxiety-like behaviours. In summary, our results indicate that mice lacking fosB are less sensitive to rewarding properties of morphine and display spatial memory impairment and suggest involvement of fosB and its proteins in motivational aspects of reinforcers as well as in learning and memory processes. PMID- 18407361 TI - Can NT-proBNP predict risk of cardiovascular mortality within 10 years? Results from an epidemiological study of elderly patients with symptoms of heart failure. AB - BACKGROUND: Heart failure has a serious prognosis. However, among elderly patients the panorama of concomitant diseases makes it difficult to implement the results from epidemiological studies. The aim of this study was to evaluate the influence of different clinical variables on cardiovascular mortality during a long-term follow-up. METHODS AND RESULTS: In all, 474 elderly patients (age 65-82 years) in primary health care were evaluated and followed during a 10 year period. All patients had symptoms associated with heart failure and were examined by a cardiologist. Blood samples including NT-proBNP were analyzed, and ECG and Doppler echocardiography were assessed. Both the systolic and diastolic function was evaluated. Functional capacity was evaluated according to the NYHA classification. During the 10 years of follow-up those with the highest quartile of plasma concentration of NT-proBNP had almost four times increased risk of cardiovascular mortality. Impaired systolic function, diabetes and reduced functional capacity were all markers of increased risk of cardiovascular mortality. All variables were also evaluated after 5 years, with higher risk ratios for a majority of variables. CONCLUSION: In this study 474 patients with symptoms of heart failure were followed during 10 years. High plasma concentration of NT-proBNP could predict almost four times increased risk of cardiovascular mortality up to 10 years. Also, impaired cardiac function according to echocardiography, and reduced functional capacity as well as diabetes all had influence on risk of cardiovascular mortality up to 10 years. PMID- 18407362 TI - Prevalence of Tako-Tsubo Syndrome among patients with suspicion of acute coronary syndrome referred to our centre. AB - BACKGROUND: The Tako-Tsubo Syndrome is a clinical entity characterized by acute but rapidly reversible left ventricular systolic dysfunction and triggered by emotional or psychological stress. The aim of our study was to determine the prevalence and characteristics of this syndrome among the patients presenting to our Centre with suspicion of acute coronary syndrome. METHODS AND RESULTS: Over a 12-month period (May 2006 to April 2007), among 82 patients referred to our catheterization laboratory with suspicion of acute coronary syndrome, 4 confirmed Tako-Tsubo Syndrome (prevalence 4.87%). The patients referred to our Centre came from Foggia's province above all. The mean age of the population was 65.5 +/- 18.48 years (range 49 to 82), with a ratio of men to women of 1:3. The syndrome characterized by acute chest pain with ST-segment elevation, absence of significant lesions in each of the 3 epicardial coronary arteries by angiography, systolic dysfunction (ejection fraction 35 +/- 9.12%) with abnormal wall motion of the mid and distal LV and hyperkinesia of the basal LV, and emotional or psychological stress immediately preceding the cardiac events. Among markers of cardiac necrosis, only serum Troponin-I increased in each patients without significant elevation of CPK and with mild elevation of CK-mb and LDH. 2 patients developed hemodynamic instability. Each patient survived with normalized ejection fraction (54.25 +/- 5.05%) and rapid restoration to previous functional cardiovascular status within 4 weeks. CONCLUSIONS: A reversible cardiomyopathy triggered by emotional or psychological stress occurs in elderly women above all and mimic acute coronary syndrome. The diagnosis of Tako-Tsubo Syndrome is based mainly on coronary and left ventricular angiography, which excludes the diagnosis of coronary artery disease and recognizes the pattern of wall-motion abnormalities. The different epidemiology of this Syndrome reported in literature demonstrates which this cardiomyopathy is underdiagnosed. PMID- 18407363 TI - Dynamic time warping: a new method in the study of poor handwriting. AB - Poor handwriting is a diagnostic criterion for developmental coordination disorder. Typical of poor handwriting is its low overall quality and the high variability of the spatial characteristics of the letters, usually assessed with a subjective handwriting scale. Recently, Dynamic Time Warping (DTW), a technique originally developed for speech recognition, was introduced for pattern recognition in handwriting. The present study evaluates its application to analyze poor handwriting. Forty children attending Dutch mainstream primary schools were recruited and based on their scores on the Concise Evaluation Scale for Children's Handwriting (Dutch abbreviation: BHK), 20 good and 20 poor writers (of whom 13 were scheduled for handwriting intervention) were identified. The groups were matched for age (7-9 years), school grade (grades 2 and 3) and handedness. The children subsequently wrote sequences of the letter "a" on a graphics tablet in three conditions (normal, fast, and accurate). Classical kinematics were obtained and for each individual letter DTW was used to calculate the distance from the mean shape. The DTW data revealed much higher variability in the letter forms of the poor writers that was independent of the kinematic results of larger trajectories, faster movements, and higher pen pressure. The current results suggest that DTW is a valid and objective technique for letter form analysis in handwriting and may hence be useful to evaluate the rehabilitation treatments of children suffering from poor handwriting. In education research it may be exploited to explore how children (should) learn to write. PMID- 18407364 TI - Determination of editors at the novel A-to-I editing positions. AB - A-to-I RNA editing modifies a variety of biologically important mRNAs, and is specifically catalyzed by either adenosine deaminase acting on RNA type 1 (ADAR1) or type 2 (ADAR2) in mammals including human. Recently several novel A-to-I editing sites were identified in mRNAs abundantly expressed in mammalian organs by means of computational genomic analysis, but which enzyme catalyzes these editing sites has not been determined. Using RNA interference (RNAi) knockdowns, we found that cytoplasmic fragile X mental retardation protein interacting protein 2 (CYFIP2) mRNA had an ADAR2-mediated editing position and bladder cancer associated protein (BLCAP) mRNA had an ADAR1-mediated editing position. In addition, we found that ADAR2 forms a complex with mRNAs with ADAR2-mediated editing positions including GluR2, kv1.1 and CYFIP2 mRNAs, particularly when the editing sites were edited in human cerebellum by means of immunoprecipitation (IP) method. CYFIP2 mRNA was ubiquitously expressed in human tissues with variable extents of K/E site editing. Because ADAR2 underactivity may be a causative molecular change of death of motor neurons in sporadic amyotrophic lateral sclerosis (ALS), this newly identified ADAR2-mediated editing position may become a useful tool for ALS research. PMID- 18407365 TI - Significance of the deep layers of entorhinal cortex for transfer of both perirhinal and amygdala inputs to the hippocampus. AB - In the rat, a number of sensory modalities converge in the perirhinal cortex (PC). The neural pathway from the perirhinal cortex to the entorhinal cortex (EC) is considered one of the main routes into the entorhinal-hippocampal network. Evidence accumulated recently suggests that EC and PC, far from being passive relay stations, actively gate impulse traffic between neocortex and hippocampus. Using slice preparation maintaining the neurocircuit connecting PC, EC, hippocampal formation and amygdala, we investigated the associative function of PC and EC with respect to sensory and motivational stimuli and the influence of the association on the neurocircuit. In horizontal slices located ventrally to the rhinal sulcus, where we stimulated area 35 and the lateral amygdala, both inputs can be independently conveyed to the dentate gyrus. In slightly more dorsal slices where we stimulated area 36 and the lateral amygdala, the coincidence of the two inputs was needed to activate the hippocampus. This need for association of the two inputs was apparently mediated by the deep layer of EC. In all instances activation of the deep layers of EC was sufficient to activate the dentate gyrus, suggesting the relevance of the deep layers in cortico-hippocampal interactions. PMID- 18407366 TI - Diversity of stress responses in dairy thermophilic streptococci. AB - Exponential and stationary phase cells of 56 strains of the dairy Streptococcus species S. thermophilus, S. macedonicus and S. salivarius, were exposed to acid, osmotic, oxidative and heat stresses to investigate the diversity of their responses. Three strains of dairy related streptococci, Lactococcus lactis ATCC11454, Enterococcus faecium DSM20477 and Enterococcus faecalis DSM20478, were included for comparison purposes. Acid and heat adaptation and cross-protection to stress were studied in ten strains with different stress response patterns. Cell death and the changes in protein expression were evaluated by plate counts and Sodium Dodecyl Sulfate Polyacrilamide Gel Electrophoresis, respectively. All strains of all species were highly tolerant of osmotic stress. With a few exceptions, acid and oxidative treatments reduced the number of viable cells by >5 log units but responses to heat stress were more variable. For some, but not all strains, stationary phase cells were more resistant to some or all stresses. Matrix cluster analysis was used to group strains on the basis of their pattern of stress response in seven clusters. Significant associations between the sources of strains and stress resistance were found for acid and oxidative stresses. Adaptation to stress during the exponential phase enhanced the survival of acid and heat stressed cells from 1 to 60,000-folds, but a detrimental effect of adaptation on cell viability was evident for oxidative and osmotic stresses for three strains. Adaptation and entry into the stationary phase resulted in significant changes of protein bands whose estimated molecular masses corresponded with those of proteins (DnaK, GprE, GroEL, and GroES) involved in the general stress response but no statistically significant correlation between stress response and band intensity was evident. PMID- 18407367 TI - Association between endogenous feline leukemia virus loads and exogenous feline leukemia virus infection in domestic cats. AB - Recently, we demonstrated that endogenous feline leukemia virus (enFeLV) loads may vary among cats of different populations and that FeLV-infected cats have higher enFeLV loads than uninfected cats. Thus, we hypothesized that enFeLV might influence the pathogenesis and outcome of FeLV infection. No significant difference in the infection outcome (regressive versus progressive infection) was observed between groups of cats with high or low enFeLV loads following FeLV-A challenge. However, cats with high enFeLV loads showed higher viral replication (plasma viral RNA and p27 antigen levels) than cats with low enFeLV loads in the early phase of the infection. The enFeLV transcription level varied at different time points, but no clear-cut pattern was observed. In conclusion, our results demonstrated an association between enFeLV loads and FeLV replication but not outcome of infection. enFeLV should be considered as an important confounder in experimental FeLV infection or vaccination studies. PMID- 18407368 TI - The impact of the chronic disease assistance plan (CDAP) on the control of type 2 diabetes in Trinidad. AB - OBJECTIVES: (1) To measure the proportion of participants who used hypoglycaemic agents provided by CDAP only, for at least 6 months, and attained an HbA1c of C, Arg194Trp, Arg280His and Arg399Gln) and the susceptibility to lung cancer among women nonsmokers, we performed a hospital based case-control study of 350 patients with newly diagnosed lung cancer and 350 cancer-free controls, frequency matched by age. Our results showed that exposure to cooking oil fume was associated with increased risk of lung cancer in Chinese women nonsmokers [odds ratio (OR)=2.51, 95% confidence interval (CI) [1.80-3.51], P<0.001]. Individuals with homozygous XRCC1 399Gln/Gln genotype (OR=1.75, 95% CI [1.02-3.01]) and XRCC1 -77 combined TC and CC genotype (OR=1.66, 95% CI [1.13 2.42]) showed a slightly higher risk for lung cancer overall. In the subgroup of adenocarcinoma cases, adjusted ORs were increased for individuals with homozygous XRCC1 399Gln/Gln genotype (OR=2.62, 95% CI [1.44-4.79]) and XRCC1 -77 combined TC and CC genotype (OR=1.85, 95% CI [1.19-2.86]). Haplotype analysis showed that T Trp-Arg-Gln haplotypes were associated with an increased risk of lung cancer among women nonsmokers (OR=2.26, 95% CI [1.38-3.68]), however, we did not observe a statistically significant joint effect of cooking oil fume and 399Gln or -77C variant allele on lung cancer among women nonsmokers. In conclusion, XRCC1 Arg399Gln and T-77C polymorphisms may alter the risk of lung cancer in women nonsmokers in China. PMID- 18407371 TI - Relationship between mass-flux reduction and source-zone mass removal: analysis of field data. AB - The magnitude of contaminant mass-flux reduction associated with a specific amount of contaminant mass removed is a key consideration for evaluating the effectiveness of a source-zone remediation effort. Thus, there is great interest in characterizing, estimating, and predicting relationships between mass-flux reduction and mass removal. Published data collected for several field studies were examined to evaluate relationships between mass-flux reduction and source zone mass removal. The studies analyzed herein represent a variety of source-zone architectures, immiscible-liquid compositions, and implemented remediation technologies. There are two general approaches to characterizing the mass-flux reduction/mass-removal relationship, end-point analysis and time-continuous analysis. End-point analysis, based on comparing masses and mass fluxes measured before and after a source-zone remediation effort, was conducted for 21 remediation projects. Mass removals were greater than 60% for all but three of the studies. Mass-flux reductions ranging from slightly less than to slightly greater than one-to-one were observed for the majority of the sites. However, these single-snapshot characterizations are limited in that the antecedent behavior is indeterminate. Time-continuous analysis, based on continuous monitoring of mass removal and mass flux, was performed for two sites, both for which data were obtained under water-flushing conditions. The reductions in mass flux were significantly different for the two sites (90% vs. approximately 8%) for similar mass removals ( approximately 40%). These results illustrate the dependence of the mass-flux-reduction/mass-removal relationship on source-zone architecture and associated mass-transfer processes. Minimal mass-flux reduction was observed for a system wherein mass removal was relatively efficient (ideal mass-transfer and displacement). Conversely, a significant degree of mass-flux reduction was observed for a site wherein mass removal was inefficient (non-ideal mass-transfer and displacement). The mass-flux-reduction/mass-removal relationship for the latter site exhibited a multi-step behavior, which cannot be predicted using some of the available simple estimation functions. PMID- 18407372 TI - Interactions between viruses and goethite during saturated flow: effects of solution pH, carbonate, and phosphate. AB - Metal oxides have great potential for controlling the fate and transport of viruses in the subsurface and water-treatment systems. The processes, however, are subject to solution chemistry. In this study, a number of column experiments were conducted to examine the effects of solution pH and anions (carbonate and phosphate) on attachment, transport, and inactivation of two bacteriophages (phiX174 and MS-2) in goethite-coated sand medium. Removal of both viruses on goethite-coated sand increased as solution pH decreased from 9.3 to 7.5, due mostly to virus inactivation. MS-2, a relatively hydrophobic virus with a lower isoelectric point (3.9), was more sensitive to the change of solution pH than phiX174, a relatively hydrophilic virus with a higher isoelectric point (6.6), in terms of their attachment and inactivation on goethite. About 90% of the MS-2 particles removed by goethite (accounting for 81% of the total input) were inactivated at pH 7.5, whereas all of the removed MS-2 particles (accounting for 10% of the total input) still remained infectious at pH 9.3. In comparison, approximately 74% of the goethite-bound phiX174 particles (accounting for 95% of the total input) lost their infectivity at pH 7.5, in contrast to a complete recovery at pH 9.3 (accounting for 65% of the total input) when the columns were eluted using a beef extract solution (pH 9.5). Presence of phosphate (20 mM H(2)PO(4)(-)) in input solution reduced virus attachment and appeared to protect the viruses from being inactivated during transport; this effect was more significant on MS-2 than on phiX174. Specifically, approximately 29% of the phiX174 particles and approximately 49% of MS-2 particles injected into the column were removed during transport. Mass recovery data showed that no phiX174 was inactivated in the presence of phosphate, whereas about 38% of the MS-2 particles attached on goethite lost their infectivity. Conversely, presence of carbonate on goethite increased virus attachment and inactivation due to contribution of additional attachment sites from protonated surface groups of the carbonate ions that were adsorbed on goethite. About 70% of the total input viruses (both phiX174 and MS-2) were removed during transport, of which 35% phiX174 and 85% MS-2 were eventually inactivated. PMID- 18407373 TI - The error of counting "errors". PMID- 18407374 TI - Are the uninsured responsible for the increase in emergency department visits in the United States? AB - STUDY OBJECTIVE: The rise in emergency department (ED) use in the United States is frequently attributed to increased visits by the uninsured. We determine whether insurance status is associated with the increase in ED visits. METHODS: Using the national Community Tracking Study Household Surveys from 1996 to 1997, 1998 to 1999, 2000 to 2001, and 2003 to 2004, we determined for each period the proportion of reported adult ED visits according to insurance status, family income, usual source of care, health status, and outpatient (non-ED) visits. Trends over time were tested for statistical significance. RESULTS: The proportion of adult ED visits by persons without insurance was stable across the decade. Uninsured individuals accounted for 15.5% of ED visits in 1996 to 1997, 16.1% in 1998 to 1999, 15.2% in 2000 to 2001, and 14.5% of visits in 2003 to 2004 (P for trend=.43). The proportion of visits by persons whose family income was greater than 400% of the federal poverty level increased from 21.9% to 29.0% (P=.002). The proportion of visits by those whose usual source of care was a physician's office increased from 52.4% in 1996 to 1997 to 59.0% in 2003 to 2004 (P=.002), whereas the proportion of visits by those without a usual source of care was essentially unchanged (9.7% of visits in 1996 to 1997 and 9.6% in 2003 to 2004; P=.74). CONCLUSION: The rise in ED visits between 1996 and 2003 cannot be primarily attributed to the uninsured. Major contributors to increasing ED utilization appear to be disproportionate increases in use by nonpoor persons and by persons whose usual source of care is a physician's office. PMID- 18407375 TI - Tort claims and adverse events in emergency medical services. AB - STUDY OBJECTIVE: Emergency medical services (EMS) provide care to acutely ill or injured patients in settings less controlled than other health care environments. Although reports describing individual EMS adverse events exist, few broader descriptions exist. The objective of the study is to characterize the types, frequencies, and outcomes of adverse events associated with insurance tort claims against EMS providers. METHODS: We performed a retrospective review of insurance liability claims from a national insurer of EMS agencies. We studied closed and open insurance liability claims from January 1, 2003, to December 31, 2004, arising from EMS response to or provision of patient care and associated with injury to patients or other individuals. We excluded events associated with employee injuries only, events with property or vehicle damage only, and emergency vehicle crashes with less than $10,000 in actual or predicted total incurred costs. We identified the category of the adverse event, the characteristics of the treating emergency units, the injured individuals, the associated injuries, and the estimated or actual total incurred costs. RESULTS: Among 326 claims included in the analysis, adverse events included emergency vehicle crash or movement (n=122; 37%; 95% confidence interval [CI] 32% to 43%), patient handling (n=118; 36%; 95% CI 31% to 41%), clinical management (n=40; 12%; 95% CI 9% to 16%), response or transport events (n=25, 8%; 95% CI 5% to 11%), and other events (n=33; 10%; 95% CI 7% to 14%). Associated injuries included death (n=54; 17%; 95% CI 13% to 21%), life-threatening or disabling injuries (n=25; 8%, 95% CI 5% to 11%), and non-life-threatening or other injuries (n=247; 76%; 95% CI 71% to 80%). The median estimated total incurred cost was $17,000 (interquartile range $7,000 to $42,000). CONCLUSION: Emergency vehicle crashes and patient handling mishaps were the most common adverse events associated with tort claims against EMS agencies. Clinical management and other incidents were less common. This effort highlights potential areas for improving EMS operations and care. PMID- 18407376 TI - Comparison of two methods of pediatric resuscitation and critical care management. AB - STUDY OBJECTIVE: We compare time to drug delivery and the incidence of dosage error between 2 different systems of medication administration: The Broselow Pediatric Emergency Tape and a standardized volume/weight-based dose reformulation of resuscitation and critical care medications (reformulated to 0.1 mL/kg). METHODS: This was a randomized crossover trial, in which volunteers (n=16) from emergency department (ED) pediatric resuscitation teams from the ED of a large, urban, teaching hospital in Australia were assigned to manage simulated (Advanced Pediatric Life Support scenario) patients. The volunteers were each presented with 3 case scenarios (brady-asystolic arrest, status epilepticus, and rapid sequence intubation requiring administration of 4, 5, and 4 medications, respectively). The order of presentation was randomized for the 2 methods. The volunteers were then asked to manage 3 case scenarios using one and then the other method (resulting in a total of 6 cases managed per participant). The dosage of each medication ordered, as well as the time to the simulated administration of that medication, was recorded for all scenarios. The expected dosages were compared with the actual dosages delivered to determine which system provided greater accuracy in medication administration. Statistical analysis was undertaken using the Wilcoxon signed rank test and McNemars test for paired proportions. RESULTS: Compared with the Broselow tape, the standardized volume/weight-based dose reformulation significantly reduced median time to medication delivery for all clinical scenarios (147 versus 72 seconds; 197 versus 87 seconds; 146 versus 64 seconds; P<.001). The proportion of dosing errors with Broselow tape across the 3 scenarios was greater than with volume/weight-based dosing (0.08 versus 0, 0 versus 0, and 0.08 versus 0.02, respectively). CONCLUSION: Use of a standardized volume/weight-based dose reformulation method is a simple, fast, and accurate method of medication delivery for the pediatric patient that eliminates the need for memorization and/or calculation. The standardized volume/weight-based dose reformulation method performs better than the Broselow tape in speed of delivery of medications used for pediatric resuscitation and critical care without any reduction in dosing accuracy. PMID- 18407377 TI - Reciprocal inhibition of G-protein signaling is induced by CB(1) cannabinoid and GABA(B) receptor interactions in rat hippocampal membranes. AB - Cannabinoid CB(1) and the metabotropic GABA(B) receptors have been shown to display similar pharmacological effects and co-localization in certain brain regions. Previous studies have reported a functional link between the two systems. As a first step to investigate the underlying molecular mechanism, here we show cross-inhibition of G-protein signaling between GABA(B) and CB(1) receptors in rat hippocampal membranes. The CB(1) agonist R-Win55,212-2 displayed high potency and efficacy in stimulating guanosine-5'-O-(3 [(35)S]thio)triphosphate, [(35)S]GTPgammaS binding. Its effect was completely blocked by the specific CB(1) antagonist AM251 suggesting that the signaling was via CB(1) receptors. The GABA(B) agonists baclofen and SKF97541 also elevated [(35)S]GTPgammaS binding by about 60%, with potency values in the micromolar range. Phaclofen behaved as a low potency antagonist with an ED(50) approximately 1mM. However, phaclofen at low doses (1 and 10nM) slightly but significantly attenuated maximal stimulation of [(35)S]GTPgammaS binding by the CB(1) agonist R Win55,212-2. The observation that higher concentrations of phaclofen had no such effect rule out the possibility of its direct action on CB(1) receptors. The pharmacologically inactive stereoisomer S-Win55,212-3 had no effect either alone or in combination with phaclofen establishing that the interaction is stereospecific in hippocampus. The specific CB(1) antagonist AM251 at a low dose (1 nM) also inhibited the efficacy of G-protein signaling of the GABA(B) receptor agonist SKF97541. Cross-talk of the two receptor systems was not detected in either spinal cord or cerebral cortex membranes. It is speculated that the interaction might occur via an allosteric interaction between a subset of GABA(B) and CB(1) receptors in rat hippocampal membranes. Although the exact molecular mechanism of the reciprocal inhibition between CB(1) and GABA(B) receptors will have to be explored by future studies it is intriguing that the cross-talk might be involved in balance tuning the endocannabinoid and GABAergic signaling in hippocampus. PMID- 18407378 TI - Promising carboranylquinazolines for boron neutron capture therapy: synthesis, characterization, and in vitro toxicity evaluation. AB - Novel classes of structurally different boronated quinazolines were designed bearing 22-37% boron by weight for potential application in BNCT of tumors. Firstly, the o-carborane cage was linked to quinazoline at C-2 position via thioether linker: 2-S-(1,2-dicarba-closo-dodecaboran(12)-1-ylmethyl)-3 phenylquinazolin-4(3H)-one. Secondly, the o-carborane cage connected to quinazoline moiety at C-4 position through an ether linkage: 4-O-(o-carboran-1 ylmethyl)-2-methylquinazoline. Finally, carborane moieties were also linked to the C-6 position of quinazoline: 6-[N-{3-(2-methyl-1,2-dicarba-closo dodecaboran(12)-1-yl)methyl}benzylidinamino]quinazolin-4(3H)-one and 6-[N-{3,5 di(2-methyl-1,2-dicarba-closo-dodecaboran(12)-1 yl)methyl}benzylidinamino]quinazolin-4(3H)-one. The water solubility was achieved by the degradative conversion of the o-carboranylquinazolines to the corresponding potassium nido-carboranyl quinazolines: 2-S-(1,2-dicarba-nido undecacarborate-1-ylmethyl)-3-phenylquinazolin-4(3H)-one, 4-O-(1,2-dicarba-nido undecacarborate-1-ylmethyl)-2-methylquinazoline, 6-[N-{3-(2-methyl-1,2-dicarba nido-undecacarborate-1-yl)methyl}benzylidinamino]quinazolin-4(3H)-one and 6-[N {3,5-di(2-methyl-1,2-dicarba-nido-undecacarborate-1 yl)methyl}benzylidinamino]quinazolin-4(3H)-one. The products were confirmed by NMR, elemental analysis, IR, and mass spectrometry. In vitro toxicity was performed with B16 melanoma cells and showed that the connection of hydrophilic nido-carborane to quinazoline moiety decreases the compound's toxicity. This cytotoxicity effect was not observed in the nido-carborane containing two cluster units which was relatively nontoxic and did not inhibit colony formation up to concentrations of 300microg boron ml(-1). The compounds described here can be considered as new candidates for BNCT. PMID- 18407379 TI - Mechanistic aspects of benzothiazepines: a class of antiarrhythmic drugs. AB - The authors have presented ab initio Hartree Fock calculations coupled with intermolecular interaction calculations to study mechanistic aspects of benzothiazepine class of calcium channel blockers. A channel model has been taken containing pore region glutamates and all three classes' sensing residues. Benzothiazepine drugs have been docked in and ternary complex (that is, drug ...Ca(2+)... channel model) stability has been studied and related to mechanistic aspects of these drugs. PMID- 18407380 TI - [Initial pattern of elderly patients and effect on their orientation after their visit in the emergency departments. Results from a national study]. AB - PURPOSE: We have little information on the geriatric characteristics of elderly patients visiting the emergency departments (ED) in France. In order to develop an adapted prevention of the arrival of some elderly patients, the determinants of their arrival to the ED deserves to be better known. METHODS: A one-day cross sectional study was conducted in French ED. A standardized questionnaire was used for each patient over 80 years (Pts), specifying the sociodemographic characteristics, the circumstances of visit to and care received in the ED, and the orientation of the patients after consulting the ED. RESULTS: On a sample of 1298 Pts, health event leading to ED started at home in 63.8% of cases. When the patients initiated themselves the recourse to health care (RHC), they called less often a general practitioner (61.9% of cases) than when the RHC was triggered by their family (69.6%, p=0.01). When a health care professional initiated the RHC, it was a GP in more than 80% of cases. Return to residence was more frequent when the patient triggered the RHC (34.5% versus 22.9% for the family and 16.0% for the professional health care, p<0,001). CONCLUSION: The actor of the decision of arrival to the ED has an impact in the RHC, in resources utilisation, and on the patient's orientation after coming in ED. The results of this study may help to design strategies aiming at avoiding unnecessary ED consultations of elderly persons. PMID- 18407381 TI - [Extensive portocava thrombosis revealing a primary antiphospholipid syndrome: a case report]. AB - We report a 20-year-old woman who presented with a massive portal thrombosis that rapidly extended to the superior and inferior vein cava system causing an acute Budd-Chiari syndrome. The investigations concluded to a primary antiphospholipid syndrome without any other prothrombotic factors. The outcome was fatal, 18 months later, despite anticoagulation, with hepatorenal syndrome and severe liver failure. PMID- 18407382 TI - Safety and reactogenicity profile of an adjuvanted H5N1 pandemic candidate vaccine in adults within a phase III safety trial. AB - A multicentre, randomized, phase III clinical trial in 5071 healthy adults was conducted to evaluate the safety and reactogenicity of a 15 microg HA dose of a candidate oil-in-water emulsion-based adjuvant system (AS)-adjuvanted split virion H5N1 (AS-H5N1) vaccine compared to a licensed seasonal influenza vaccine, Fluarix.(1) Stringent criteria were used to evaluate adverse events and reactogenicity profile. Overall, 96.7% of the 5071 vaccinated subjects completed the study. Significantly more participants in the AS-H5N1 vaccine group reported general or local adverse events. Pain was the most common symptom in both treatment groups. Less than 1% of subjects withdrew from the study due to adverse events and no withdrawals were due to serious adverse events related to vaccination. The safety and reactogenicity profile of the AS-H5N1 candidate vaccine can be considered clinically acceptable in the context of its use against pandemic influenza. PMID- 18407383 TI - Evaluation of a vectored equine herpesvirus type 1 (EHV-1) vaccine expressing H3 haemagglutinin in the protection of dogs against canine influenza. AB - In 2004, canine influenza virus (CIV) was identified as a respiratory pathogen of dogs for the first time and found to be closely related to H3N8 equine influenza virus (EIV). We generated a recombinant vectored vaccine that expresses H3 of a recent isolate of EIV using equine herpesvirus type 1 (EHV-1) as the delivery vehicle. This EHV-1 vectored vaccine exhibited robust and stable EIV H3 expression and induced a strong influenza virus-specific response in both mice and dogs upon intranasal or subcutaneous administration. Furthermore, upon challenge with the recent CIV isolate A/canine/PA/10915-07, protection of vaccinated dogs could be demonstrated by a significant reduction in clinical sings, and, more importantly, by a significant reduction in virus shedding. We concluded that the EHV-1/H3 recombinant vector can be a valuable alternative for protection of dogs against clinical disease induced by CIV and can significantly reduce virus spread. PMID- 18407384 TI - Influenza vaccine dosages. PMID- 18407385 TI - Influenza vaccination among primary healthcare workers. AB - Influenza vaccination of health care workers (HCW) is widely recommended, but immunization rates are low. In order to assess performance among primary HCW and identify barriers, we conducted a survey among the staff of 27 primary care community clinics in Jerusalem. The low rate of vaccination (30.2%) was in contrast with the large proportion of HCW who considered it desirable to immunize primary health clinic staff (72.1%). Physicians reported having been immunized significantly more (p=0.008) than the rest of the staff. They also had better knowledge and more favorable attitude towards immunization. Also associated (independently of profession) with performance of immunization were age (p<0.001), knowledge (that immunization can not cause influenza, p=0.051), attitude (the belief that it is desirable to immunize primary HCW, p<0.001), previous years' performance of immunization (p<0.001) and a physician's recommendation (p=0.042). A media scare which occurred during the vaccination period was reported to have influenced the decision not to get immunized of 34.1% of HCW who had not been immunized. The study results raise hope of prospective increase in vaccination through educational and technical interventions and by increasing physician involvement. PMID- 18407386 TI - Two versus three doses of a meningococcal C conjugate vaccine concomitantly administered with a hexavalent DTaP-IPV-HBV/Hib vaccine in healthy infants. AB - The immunogenicity and reactogenicity of a meningococcal serogroup C (MenC) conjugate vaccine given concomitantly with DTaP-IPV-HBV/Hib vaccine according to a two- or three-dose schedule in healthy infants was evaluated. At 1 month post vaccination, 98% (two doses) and 100% (three doses) of subjects had serum bactericidal antibody using human complement assay (hSBA) titres > or =1:8; at 12 months of age > or =89% of subjects in each group remained seroprotected. Induction of immunological memory, as evaluated by administration of a meningococcal serogroup A/C polysaccharide vaccine challenge dose, was similar for both regimens and no interference was observed in the immune response to MenC or hepatitis B virus antigens. Reactogenicity was similar in each group. MenC conjugate vaccine given concomitantly with DTaP-IPV-HBV/Hib to healthy infants in the first year of life using a two-dose schedule is as safe and immunogenic as a three-dose regimen. PMID- 18407387 TI - Gait initiation in older adults with postural instability. AB - BACKGROUND: This study examined changes in the translation of the center of pressure during forward and lateral (90 degrees to the side) gait initiation in two populations of older adults with postural instability. METHODS: Twenty-eight older adults transitioning to frailty and 16 persons with Parkinson's disease in the "on medication state" were evaluated during initiation trials. Displacements, velocities, and smoothness of the center of pressure trace were calculated and compared. FINDINGS: Both groups produced movements of the center of pressure that on average were reduced compared to healthy populations. Adults transitioning to frailty were able to scale the output of the motor program so forces that propel the body in the intended direction of movement were maximized as evidenced by movements of the center of pressure. The adults transitioning to frailty produced patterns of center of pressure trajectories that were more similar to healthy adults where as individuals with Parkinson's disease produced trajectories that were counterproductive to producing efficient gait initiation in both the forward and lateral direction. INTERPRETATION: These findings suggest that persons with Parkinson's disease even when in the medicated state exhibit inefficient postural adjustments during both forward and lateral gait initiation and that these postural adjustments are more susceptible to deterioration from the complex interaction of central and peripheral changes associated with Parkinson's disease than to aging alone. PMID- 18407388 TI - Moving toward effective ozone flux assessment. AB - We present a comment about "Ozone risk assessment for plants: central role of metabolism-dependent changes in reducing power" by Dizengremel, Le Thiec, Bagard, and Jolivet. As tools for summarizing plant O(3) sensitivity in simple indices, Dizengremel et al. suggest: reducing power, as antioxidant regeneration through the Halliwell/Asada cycle requires NADPH from the photosynthetic light reaction; Rubisco/PEPc ratio, as an index of the energy balance between anabolic and catabolic reactions; and water-use efficiency as a time-integrated approximation of the carbon gain to stomatal O(3) uptake ratio. The scientific background is solid, and simple enough (although expensive) to be translated into modelling and routine use. In the last decade, several approaches have been developed, mostly by using photosynthesis as a metric of defence. All these approaches should be experimentally tested in different and realistic conditions, before the results are transferred to the field and used in effective O(3) flux modelling and assessment. PMID- 18407389 TI - Comparison of total mercury and methylmercury cycling at five sites using the small watershed approach. AB - The small watershed approach is well-suited but underutilized in mercury research. We applied the small watershed approach to investigate total mercury (THg) and methylmercury (MeHg) dynamics in streamwater at the five diverse forested headwater catchments of the US Geological Survey Water, Energy, and Biogeochemical Budgets (WEBB) program. At all sites, baseflow THg was generally less than 1ng L(-1) and MeHg was less than 0.2ng L(-1). THg and MeHg concentrations increased with streamflow, so export was primarily episodic. At three sites, THg and MeHg concentration and export were dominated by the particulate fraction in association with POC at high flows, with maximum THg (MeHg) concentrations of 94 (2.56)ng L(-1) at Sleepers River, Vermont; 112 (0.75)ng L(-1) at Rio Icacos, Puerto Rico; and 55 (0.80)ng L(-1) at Panola Mt., Georgia. Filtered (<0.7microm) THg increased more modestly with flow in association with the hydrophobic acid fraction (HPOA) of DOC, with maximum filtered THg concentrations near 5ng L(-1) at both Sleepers and Icacos. At Andrews Creek, Colorado, THg export was also episodic but was dominated by filtered THg, as POC concentrations were low. MeHg typically tracked THg so that each site had a fairly constant MeHg/THg ratio, which ranged from near zero at Andrews to 15% at the low-relief, groundwater-dominated Allequash Creek, Wisconsin. Allequash was the only site with filtered MeHg consistently above detection, and the filtered fraction dominated both THg and MeHg. Relative to inputs in wet deposition, watershed retention of THg (minus any subsequent volatilization) was 96.6% at Allequash, 60% at Sleepers, and 83% at Andrews. Icacos had a net export of THg, possibly due to historic gold mining or frequent disturbance from landslides. Quantification and interpretation of Hg dynamics was facilitated by the small watershed approach with emphasis on event sampling. PMID- 18407390 TI - Successful re-exposure to clozapine following uneventful rechallenge with olanzapine in a patient with neutropenia related to both agents. PMID- 18407391 TI - Combined antipsychotic treatment involving clozapine and aripiprazole. AB - Treatment resistance is considered a challenging problem of antipsychotic pharmacotherapy. In such cases, combination approaches are commonly used, for instance the add-on of aripiprazole to clozapine. This review aims at giving an overview of the present knowledge on this strategy. We performed a keyword-based screening of databases (including November 2007) and evaluated the data in a systematic manner. The courses of 94 patients were reported in 11 publications. At a mean dosage of 20.5 mg/day, aripiprazole achieved clinical improvement of psychotic symptoms and facilitated a dose reduction of clozapine from 476.7 to 425.1 mg/day. In parallel, clozapine serum levels decreased from 611 to 523 ng/ml. No pharmacokinetic interactions were reported, and clozapine-induced side effects ameliorated. However, single cases of extrapyramidal side effects occurred. The combination of clozapine and aripiprazole follows a neurobiological rationale and appears to be effective and tolerable. The results of placebo controlled trials might allow further insight into the benefits and risks of this strategy. PMID- 18407392 TI - Steroid and beta-adrenergic receptor modifications in target organs of broiler chickens fed with a diet containing beta2-adrenergic agents. AB - The illegal use of beta2-agonists as repartitioning agents to improve production performance and carcass composition can induce changes in various organs of exposed animals. The aim of the present study was to evaluate the effects induced by dietary beta-agonists on beta-AR, AnR and GR in male broiler target organs. Fifty-four male broiler chickens (Ross 508), 26 days old, were randomly divided into three homogeneous experimental groups and fed for 21 days with a standard diet containing placebo (group 1, control), 1ppm of clenbuterol (group 2) and 1ppm of cimaterol (group 3). Tissue samples of heart, lung, brain, testicle, spleen, thymus and Bursa of Fabricius were collected post-mortem then cytosol fractions were used for AnR (testicles) and GR (spleen, thymus and Bursa of Fabricius), and membrane fractions for beta-AR (all tissues but testicles) determination by binding assays. The dietary administration of beta-adrenergic agents as repartitioning agents induced a significant decrease in AnR concentration in the testicle, in GR levels in the lymphoid tissues and in beta AR concentrations of different target organs of male chickens. Present data confirm those observed in female chickens and suggest that in poultry the regulation exerted by adrenergic stimulation on steroid receptor concentrations produces different effects than in mammals. PMID- 18407393 TI - Skin decontamination of glyphosate from human skin in vitro. AB - This study compared three model decontaminant solutions (tap water, isotonic saline, and hypertonic saline) for their ability to remove a model herbicide (glyphosate) from an in vitro human skin model. Human cadaver skin was dosed (approximately 375microg) of [14C]-glyphosate on 3cm2 per skin. After each exposure time (1, 3, and 30min post-dosing, respectively), the surface skin was washed three times (4ml per time) with each solution. After washing, the skin was stripped twice with tape discs. Lastly, the wash solutions, strippings, receptor fluid, and remainder of skin were liquid scintillation analyzer counted to determine the amount of glyphosate. There were no statistical differences among these groups at any time points. The total mass balance recovery at three time exposure points was between 94.8% and 102.4%. The wash off rates (glyphosate in wash solutions) at three different exposure times is 79-101.2%. Thus the three tested decontaminants possess similar effectiveness in removing glyphosate from skin. This in vitro model is not only economic and rapid, but also provides quantitative data that may aid screening for optimal decontaminants. PMID- 18407394 TI - Natural uranium disturbs mouse folliculogenesis in vivo and oocyte meiosis in vitro. AB - We investigated whether uranium intoxication affects female fertility by assessing its effects on ovarian function and on the oocyte. We treated two groups of female mice for 15 weeks with 5, 50 or 400 mg/L of uranyl nitrate in drinking water. In the first group, mice were euthanized immediately after intoxication. Mice of the second group were paired after intoxication with untreated males. Dams and their female pups were euthanized 3 months after the end of intoxication. We assayed the kidneys, femurs and one ovary per female for U content and collected the other ovary for histology. The number and size of all the ovarian follicles were analyzed. Mice from the first group and female pups had significantly fewer large antral follicles (O > 200 microm) than the untreated mice. By contrast, dams in the second group had more secondary and early preantral follicles (O 70-110 microm) than untreated mice. However, U had no effect on follicle atresia. We then analyzed the in vitro effects of U on oocyte maturation and fragmentation. GV-oocytes were cultured in the presence of 1mM uranyl acetate and observed for 72 h. Oocyte maturation was slowed down by U during resumption of meiosis and at metaphase II. However, the rhythm and rate of oocyte fragmentation were similar to those of control mice. Our findings demonstrate that U induces changes in folliculogenesis and oocyte maturation in mice and could consequently represent a risk for women who are chronically exposed. PMID- 18407395 TI - Effects of methylated chrysenes on AhR-dependent and -independent toxic events in rat liver epithelial cells. AB - Methylated chrysenes (MeChry) are important cigarette smoke constituents and 5 MeChry has been listed as possibly carcinogenic to humans. Although a major attention has been in past paid especially to mutagenic, tumor-initiating effects of MeChry, little is known about toxic effects of MeChry related to tumor promotion. As the position of methyl group has been repeatedly observed to determine genotoxic effects of MeChry, we examined both genotoxic and nongenotoxic effects of MeChry, using rat liver cell lines as experimental models. All six MeChry were relatively efficient aryl hydrocarbon receptor (AhR) agonists, with 3- and 6-MeChry being the most potent inducers of the AhR-mediated reporter gene activity. All six compounds disrupted contact inhibition in rat liver epithelial WB-F344 cells, a process previously reported to be AhR dependent, suggesting that MeChry may interfere with cell cycle control in an AhR dependent manner. In contrast, only 5- and 6-MeChry were found to acutely inhibit gap junctional intercellular communication (GJIC), another parameter correlating with tumor promoting effects of xenobiotics. Both 5- and 6-MeChry were efficient inducers of mRNA expression of enzymes involved in metabolic activation of polycyclic aromatic hydrocarbons, including cytochromes P450 1A1/1B1 and aldo keto reductase 1C9. However, only 5-MeChry, and not 6-MeChry, induced significant formation of DNA adducts in rat liver epithelial cells, which corresponded with its ability to induce high accumulation of cells in S-phase. On the other hand, 5 MeChry induced neither apoptosis related to DNA damage nor phosphorylation of p53 tumor suppressor. Taken together, our results suggest that methyl group position may affect both genotoxic and nongenotoxic effects of MeChry, such as formation of DNA adducts and inhibition of GJIC. All MeChry showed a potency to disrupt cell proliferation control, while 5-MeChry was a single compound inducing DNA damage, disruption of cell cycle control and inhibition of GJIC in rat liver cells. PMID- 18407396 TI - Nanoparticle-reinforced resin-based dental composites. AB - OBJECTIVE: TiO(2) nanoparticles treated with the organosilane allyltriethoxysilane (ATES) are used to improve the mechanical properties of dental resin-based composites (RBCs, Z100, 3M ESPE). METHODS: TiO(2) nanoparticles were sonically dispersed in an ethanol solution containing ATES. The modified particles were washed in pure ethanol and dried before being used as filler. Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM) were used to analyze the nanoparticles. Five groups of composite resin specimens were prepared: one control group, and four groups using either modified or unmodified nanoparticles comprising 0.5% or 1.0% of the material by weight. The mechanical properties (microhardness and flexural strength) of all five groups of specimens were measured. RESULTS: After modification, the particles' FTIR spectrum shows a new absorption doublet at 1200 and 1020 cm(-1). TEM images show that the modified particles have better dispersion, and that their clusters are small enough to create a homogeneous surface on dental RBCs. Composite resin specimens including modified nano-TiO(2) have significantly better mechanical properties than the control group (P<0.05). The improvement of adding 1.0 wt.% modified nano-TiO(2) particles was better than that of 0.5 wt.%. CONCLUSIONS: Surface modification by the organosilane ATES influences the dispersion and linkage of TiO(2) nanoparticles within a resin matrix, and the modified particles are found to improve the microhardness and flexural strength of dental RBCs. PMID- 18407397 TI - Hospital recovery following Thermachoice ablation is not dependent on setting (outpatient or daycase) or rescue analgesia: unexpected result. AB - BACKGROUND: Thermal balloon endometrial ablation (TBEA) is increasingly being performed in the outpatient setting under local anaesthesia (LA) rather than in a daycase setting under general anaesthesia (GA). Our aim was to compare the post operative rescue analgesia requirements and duration of hospital stay in women undergoing outpatient (LA) and daycase (GA) TBEA. METHODS: Prospective observational study of consecutively recruited women who underwent outpatient (LA) TBEA (n=51) and daycase (GA) TBEA (n=50) over the same time period. Analgesia that was provided additional to the standard administered analgesic regimen was considered rescue analgesia. The main outcome measures were requirement for rescue analgesia and duration of hospital stay in both cohorts. RESULT(S): LA compared to GA cohorts had shorter hospital stays (11 h [95% CI 9 13] vs. 17 h [95% CI 14-20]) and lower analgesia requirements. However, multivariate regression, correcting for all known confounders, showed that duration of stay was independent of setting for ablation or amount of rescue analgesia. CONCLUSION(S): Duration of hospital stay is not entirely dependent on whether outpatient or daycase endometrial ablation is considered. This unexpected preliminary finding deserves to be validated in future confirmatory trials that compare outpatient and daycase treatments. We also discuss the confounding factors that should be considered when designing such trials. PMID- 18407398 TI - Identification and mapping of submerged plants in a shallow lake using Quickbird satellite data. AB - Turkey is a country rich in lakes and wetlands--monitoring of all these will require advances in technology such as remote sensing. In this study, the aquatic plants of the large and shallow Lake Mogan, located in Central Anatolia were identified and mapped using high spatial resolution Quickbird imagery. As Lake Mogan is an important bird area the assessment of submerged plant species is of great value for ecosystem conservation and management. Quickbird multispectral image acquired on August 6, 2005 was geometrically corrected and a water mask was used based on strong absorption of Near Infrared (NIR) wavelengths by calm, clear and deep water. The water mask was applied using band reflectance values for a specific pixel satisfying the conditions of band decreasing property (Green>Red>NIR) and NIRC. humicola>B. subtilis. The sequence of total thermal effect (Q(total)) and growth rate constant (k) is mixed microorganisms>B. subtilis>C. humicola. These results are important to further studies of the physiology and pharmacology of C. humicola and B. subtilis and may support the theory of restoring contaminated soil. PMID- 18407412 TI - Course and prognostic factors of whiplash: a systematic review and meta-analysis. AB - We conducted a systematic review and meta-analysis of prospective cohort studies of subjects with acute whiplash injuries. The aim was to describe the course of recovery, pain and disability symptoms and also to assess the influence of different prognostic factors on outcome. Studies were selected for inclusion if they enrolled subjects with neck pain within six weeks of a car accident and measured pain and/or disability outcomes. Studies were located via a sensitive search of electronic databases; Medline, Embase, CINAHL, Cochrane database, ACP Journal club, DARE and Psychinfo and through hand-searches of relevant previous reviews. Methodological quality of all studies was assessed using a six item checklist. Sixty-seven articles, describing 38 separate cohorts were included. Recovery rates were extremely variable across studies but homogeneity was improved when only data from studies of more robust methodological quality were considered. These data suggest that recovery occurs for a substantial proportion of subjects in the initial 3 months after the accident but after this time recovery rates level off. Pain and disability symptoms also reduce rapidly in the initial months after the accident but show little improvement after 3 months have elapsed. Data regarding the prognostic factors associated with poor recovery were difficult to interpret due to heterogeneity of the techniques used to assess such associations and the way in which they are reported. There was also wide variation in the measurement of outcome and the use of validated measures would improve interpretability and comparability of future studies. PMID- 18407413 TI - Effectiveness of leech therapy in women with symptomatic arthrosis of the first carpometacarpal joint: a randomized controlled trial. AB - Leech therapy has been shown to be effective for symptomatic treatment of osteoarthritis of the knee. We aimed to investigate the effectiveness of leech therapy in another type of osteoarthritis, osteoarthritis of the first carpometacarpal joint (thumb saddle joint). Thirty-two women with symptomatic painful osteoarthritis of the first carpometacarpal joint and who scored>40 mm on a 100mm VAS pain scale were randomized to a single treatment with 2-3 locally applied leeches (leech group) or a 30-day course with topical diclofenac twice a day. Primary outcome measure was change of overall pain (mean of VAS for pain at rest, in motion, during grip) from baseline to day 7. Secondary outcomes were functional disability (DASH-questionnaire), quality of life (QoL, SF-36) and grip strength. Patients were examined baseline and at days 7, 30 and 60 after treatment. Overall pain score at day 7 was reduced from 59.6+/-13.8 to 27.1+/ 20.6 in the leech group (n=16) and from 50.6+/-13.3 to 46.9+/-18.5 with diclofenac (n=16) (group difference -26.5, 95%CI -40.3; -12.7; p=0.0003). Group differences for pain relief favoring the leech treatment increased at days 30 and 60. Significant treatment effects were also observed for the DASH score, QoL and grip. Results were not affected by outcome expectation or consumption of analgetics. A single course of leech therapy is effective in relieving pain, improving disability and QoL for at least 2 months. The potential of leech therapy for treatment of arthritic pain and underlying mechanisms should be further investigated. PMID- 18407414 TI - NK-1 receptors in the rostral ventromedial medulla contribute to hyperalgesia produced by intraplantar injection of capsaicin. AB - The rostral ventromedial medulla (RVM) is an area of the brainstem involved in the descending modulation of nociception at the level of the spinal cord. Although the RVM is involved in the inhibition or facilitation of nociception, the underlying mechanisms are not understood. Here we examined the role of the neuropeptide substance P and neurokinin-1 (NK-1) receptors located in the RVM on withdrawal responses evoked by mechanical and heat stimuli applied to the rat hindpaw under normal conditions and during hyperalgesia produced by capsaicin. The mechanical withdrawal threshold was obtained using von Frey monofilaments applied to the plantar surface of the hindpaw. Sensitivity to heat was determined by measuring the latency to withdrawal from radiant heat applied to the plantar surface. Mechanical and heat hyperalgesia were defined as a decrease in withdrawal response threshold or latency, respectively. Rats were prepared with a chronic cannula and either vehicle or the NK-1 receptor antagonists, L-733,060 or RP-67580, was injected into the RVM. Paw withdrawal responses were obtained before and after RVM injection, and then at 5, 30, and 60 min after an intraplantar injection of capsaicin (10 microg). Injection of the NK-1 antagonists at doses of 0.5 pmol or higher did not alter withdrawal responses to mechanical or heat stimuli under normal conditions but reduced the duration of nocifensive behavior and the mechanical and heat hyperalgesia produced by capsaicin. These findings suggest that the activation of NK-1 receptors in the RVM contributes to the hyperalgesia produced by capsaicin. PMID- 18407415 TI - Momentum-resolved energy loss near edge structure in SrCu2(BO3)2. AB - In this paper, we present a detailed study of the electronic structure of the strontium copper borate SrCu(2)(BO(3))(2) using momentum-resolved energy loss near edge structure (ELNES) and ab initio band structure calculations. Information on the energy position, the character and the spatial orientation of the unoccupied states of this compound can be efficiently retrieved taking advantage of the nearly parallel illumination available in a conventional transmission electron microscope (CTEM). These results shed light on the bonding mechanisms taking place in complex oxides such as SrCu(2)(BO(3))(2). PMID- 18407416 TI - Angiostrongylus vasorum and Eucoleus aerophilus in foxes (Vulpes vulpes) in Great Britain. AB - The nematode parasite Angiostrongylus vasorum is a source of increasing concern in several parts of the world, where it causes significant disease in dogs. Wild canids, especially foxes, are likely to have a role in the epidemiology of canine infection, and the parasite could also affect fox health and population dynamics. The heart and pulmonary vasculature of 546 foxes culled mostly by gamekeepers in Great Britain in 2005-2006 were examined by dissection and a modified flushing technique. Forty foxes were found to be infected, giving an overall prevalence in the UK fox population of 7.3% (5.3-9.9). Prevalence varied widely between regions, from 0% (0-3) in Scotland and northern England to 23% (16-32) in south east England. This closely matches the perceived incidence of disease in dogs, which is commonly diagnosed in the south-east but rarely in the north. In the Midlands, where disease has recently appeared in dogs, prevalence in foxes was 4.8% (2-11). Close geographical overlap of parasite distribution in foxes and dogs does not necessarily indicate an important wildlife reservoir of infection, but does suggest that A. vasorum might be spreading northwards. The hearts of infected foxes had thicker right ventricles than those of uninfected foxes, suggesting that the parasite could affect fox health and fitness. Burdens ranged from 1 to 59 adult nematodes. Sex, age and body condition were not significantly associated with infection. Eucoleus aerophilus and Crenosoma vulpis, nematode parasites of the respiratory system, were found in 213 and 11 foxes respectively, with slightly higher prevalence of E. aerophilus in the south and east. No specimens of the heartworm Dirofilaria immitis were found, giving an upper 95% confidence interval for prevalence of 0.84%. PMID- 18407417 TI - Clostridial collagenase aggravates the systemic inflammatory response in rats with partial-thickness burns. AB - AIM: Clostridial collagenase A (CCA) has been shown effective in degrading collagen in eschar tissue and promoting healing in partial-thickness burns. As there are also reports of fever, leukocytosis, increased C-reactive protein (CRP) levels and septic complications during treatment with CCA, we aimed to determine in rats whether CCA aggravates the systemic inflammatory response. METHODS: Rats with partial-thickness burns were randomly divided into groups with either no dressing (ND), povidone-iodine dressing (PID) or CCA dressing (CCAD). Body weights and temperatures, blood leukocyte counts, and serum levels of CRP, interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-alpha), were measured at 0, 3, and 24h and days 3 and 7 from burn. Wounds were cultured on days 1, 3 and 7 and burn depth was evaluated on day 1. RESULTS: Body weights for all groups were significantly lower after burn, with highest loss (25.5%) in the CCAD group. At 3h a significant drop in rectal temperature was noted in all groups. The CCAD group had higher rectal temperature levels than the PID group on days 3 and 7 (p<0.05). Changes in serum levels of CRP, IL-1 beta, IL-6 and TNF-alpha were not significant in the ND and PID groups; the CCAD group showed a significant rise in serum levels of CRP on day 1, of IL-6 on day 3 and of TNF-alpha on day 7. Wound infection was more common in CCAD group and increased on days 3 and 7, but this was insignificant. CONCLUSION: CCA aggravated the systemic inflammatory response in rats with partial-thickness burns, which is accompanied by a higher risk of infection. PMID- 18407418 TI - Use of lyophilized bovine collagen for split-thickness skin graft donor site management. AB - Donor site management after split-thickness skin graft applications can have problems such as late healing and pain. Many dressing methods and medical applications are reported to solve these problems but none of them were ideal. In this study we aimed to promote epithelisation and remove pain earlier with using lyophilized bovine collagen (gelfix spray). According to our results, epithelisation time for the gelfix group was earlier than control group (9.09 days mean and 11.2 days mean for control group (p<0.05)). Pain relief was determined by visual analogue pain scale. In the gelfix group, there was pain relief up to 40 h from the operation. There were no differences between groups for scarring 30 and 90 days after surgery. PMID- 18407419 TI - Growth cone collapse stimulated by both calpain- and Rho-mediated pathways. AB - The signal transduction pathways regulating growth cone motility remain poorly defined. Previously, we have characterized the inhibitory molecule, motuporamine C (MotC), as a robust stimulator of growth cone collapse. Utilizing MotC as a research tool to elucidate pathways involved with collapse, we have previously shown that the Rho-Rho kinase (ROCK) pathway is partially required for collapse. In this study, we report MotC induces a high-amplitude rise in intracellular free Ca(2+) concentration levels in chicks, resulting in the activation of the Ca(2+) sensitive protease, calpain. Furthermore, we show that while calpain is necessary for collapse, inhibition of calpain only partially attenuates MotC-mediated collapse. Instead, concomitant inhibition of both the Rho-ROCK and calpain pathways has an additive effect in attenuating the collapse response to MotC. To our knowledge, this is the first demonstration of concurrent activation of calpain and Rho-ROCK signaling during growth cone collapse. Our data support a model of growth cone collapse that requires the combinatorial regulation of multiple signal transduction cascades that likely target different cellular mechanisms to induce this motile response. PMID- 18407420 TI - Tonic and stimulus-evoked nitric oxide production in the mouse olfactory bulb. AB - Nitric oxide (NO) has been long assumed to play a key role in mammalian olfaction. This was based largely on circumstantial evidence, i.e. prominent staining for nitric oxide synthase (NOS) and cyclic guanosine 3',5'-cyclic monophosphate (cGMP) or soluble guanylyl cyclase, an effector enzyme activated by NO, in local interneurons of the olfactory bulb. Here we employ innovative custom fabricated NO micro-sensors to obtain the first direct, time-resolved measurements of NO signaling in the olfactory bulb. In 400 microm thick mouse olfactory bulb slices, we detected a steady average basal level of 87 nM NO in the extracellular space of mitral or granule cell layers. This NO 'tone' was sensitive to NOS substrate manipulation (200 microM L-arginine, 2 mM N(G)-nitro-L arginine methyl ester) and Mg(2+) modulation of N-methyl-D-aspartate (NMDA) receptor conductance. Electrical stimulation of olfactory nerve fibers evoked transient (peak at 10 s) increments in NO levels 90-100 nM above baseline. In the anesthetized mouse, NO micro-sensors inserted into the granule cell layer detected NO transients averaging 55 nM in amplitude and peaking at 3.4 s after onset of a 5 s odorant stimulation. These findings suggest dual roles for NO signaling in the olfactory bulb: tonic inhibitory control of principal neurons, and regulation of circuit dynamics during odor information processing. PMID- 18407421 TI - Fibroblast growth factor 2 enhances striatal and nigral neurogenesis in the acute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine model of Parkinson's disease. AB - In response to injury, endogenous precursors in the adult brain can proliferate and generate new neurons, which may have the capacity to replace dysfunctional or dead cells. Although injury-induced neurogenesis has been demonstrated in animal models of stroke, Alzheimer's disease (AD) and Huntington's disease (HD), studies of Parkinson's disease (PD) have produced conflicting results. In this study, we investigated the ability of adult mice to generate new neurons in response to the parkinsonian toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which causes selective degeneration of nigrostriatal dopamine neurons. MPTP lesions increased the incorporation of 5-bromo-2'-deoxyuridine-5'-monophosphate (BrdU), as well as the number of cells that co-expressed BrdU and the immature neuronal marker doublecortin (DCX), in two neuroproliferative regions-the subgranular zone of the dentate gyrus (DG) and the rostral subventricular zone (SVZ). BrdU labeled, DCX-expressing cells were not found in the substantia nigra (SN) of MPTP treated mice, where neuronal cell bodies are destroyed, but were present in increased numbers in the striatum, where SN neurons lost in PD normally project. Fibroblast growth factor-2 (FGF-2), which enhances neurogenesis in a mouse model of HD, also increased the number of BrdU/DCX-immunopositive cells in the SN of MPTP-treated mice. Thus, MPTP-induced brain injury increases striatal neurogenesis and, in combination with FGF-2 treatment, also stimulates neurogenesis in SN. PMID- 18407422 TI - Alpha-synuclein-immunopositive myenteric neurons and vagal preganglionic terminals: autonomic pathway implicated in Parkinson's disease? AB - The protein alpha-synuclein is implicated in the development of Parkinson's disease. The molecule forms Lewy body aggregates that are hallmarks of the disease, has been associated with the spread of neuropathology from the peripheral to the CNS, and appears to be involved with the autonomic disorders responsible for the gastrointestinal (GI) symptoms of individuals afflicted with Parkinson's. To characterize the normative expression of alpha-synuclein in the innervation of the GI tract, we examined both the postganglionic neurons and the preganglionic projections by which the disease is postulated to retrogradely invade the CNS. Specifically, in Fischer 344 and Sprague-Dawley rats, immunohistochemistry in conjunction with injections of the tracer Dextran-Texas Red was used to determine, respectively, the expression of alpha-synuclein in the myenteric plexus and in the vagal terminals. Alpha-synuclein is expressed in a subpopulation of myenteric neurons, with the proportion of positive somata increasing from the stomach (approximately 3%) through duodenum (proximal, approximately 6%; distal, approximately 13%) to jejunum (approximately 22%). Alpha-synuclein is co-expressed with the nitrergic enzyme nitric oxide synthase (NOS) or the cholinergic markers calbindin and calretinin in regionally specific patterns: approximately 90% of forestomach neurons positive for alpha-synuclein express NOS, whereas approximately 92% of corpus-antrum neurons positive for alpha-synuclein express cholinergic markers. Vagal afferent endings in the myenteric plexus and the GI smooth muscle do not express alpha-synuclein, whereas, virtually all vagal preganglionic projections to the gut express alpha synuclein, both in axons and in terminal varicosities in apposition with myenteric neurons. Vagotomy eliminates most, but not all, alpha-synuclein positive neurites in the plexus. Some vagal preganglionic efferents expressing alpha-synuclein form varicose terminal rings around myenteric plexus neurons that are also positive for the protein, thus providing a candidate alpha-synuclein expressing pathway for the retrograde transport of putative Parkinson's pathogens or toxins from the ENS to the CNS. PMID- 18407423 TI - Receipt of vaginal-cervical stimulation modifies synapsin content in limbic areas of the female rat. AB - Female rats require a sufficient amount and pattern of vaginal-cervical stimulation to initiate neuroendocrine changes required for the successful implantation of a fertilized ovum in the uterus. These changes are characterized by twice daily prolactin surges that last 10-12 days. Following a sterile mating, the endocrine changes are still observed, and are termed pseudopregnancy (PSP). The mating stimulation required to initiate these changes prior to pregnancy or PSP has a neural representation, which we have termed the intromission mnemonic. We sought to examine if the formation of the intromission mnemonic is accompanied by alterations in the number or density of synapses in limbic areas by immuno labeling a pre-synaptic protein, synapsin. Groups of cycling female rats on proestrus day received either 15 or 5 intromissions or mounts-without intromissions from a vasectomized male; an additional time-matched control group was left in the home cage. All females were perfused after 90 min or 8 h. The brains were removed and sliced, and the amygdala and hippocampus immunostained for synapsin, then imaged by confocal microscopy. We found that 90 min after mating sufficient for PSP, the number of synapsin puncta (points of immunoreactivity equivalent to a synapse) was decreased and the intensity of the synapsin staining was increased in the posterodorsal medial amygdala (MePD). A similar reduction of puncta was observed in the CA1 region of the hippocampus, and an increase of intensity occurred in the basolateral amygdala. Spaced intromissions had no effect on synapsin expression anywhere examined. Intensity reductions unrelated to receipt of vaginal-cervical stimulation were observed in the hippocampus. None of these effects were observed after 8 h. Together, these results raise the possibility that synapses in the MePD may be pruned after mating stimulation, resulting in pathway-specific stabilization that contributes to the intromission mnemonic associated with the establishment of PSP. PMID- 18407424 TI - High titers of mucosal and systemic anti-PrP antibodies abrogate oral prion infection in mucosal-vaccinated mice. AB - Significant outbreaks of prion disease linked to oral exposure of the prion agent have occurred in animal and human populations. These disorders are associated with a conformational change of a normal protein, PrP(C) (C for cellular), to a toxic and infectious form, PrP(Sc) (Sc for scrapie). None of the prionoses currently have an effective treatment. Some forms of prion disease are thought to be spread by oral ingestion of PrP(Sc), such as chronic wasting disease and variant Creutzfeldt-Jakob disease. Attempts to obtain an active immunization in wild-type animals have been hampered by auto-tolerance to PrP and potential toxicity. Previously, we demonstrated that it is possible to overcome tolerance and obtain a specific anti-PrP antibody response by oral inoculation of the PrP protein expressed in an attenuated Salmonella vector. This past study showed that 30% of vaccinated animals were free of disease more than 350 days post-challenge. In the current study we have both optimized the vaccination protocol and divided the vaccinated mice into low and high immune responder groups prior to oral challenge with PrP(Sc) scrapie strain 139A. These methodological refinements led to a significantly improved therapeutic response. 100% of mice with a high mucosal anti-PrP titer immunoglobulin (Ig) A and a high systemic IgG titer, prior to challenge, remained without symptoms of PrP infection at 400 days (log-rank test P<0.0001 versus sham controls). The brains from these surviving clinically asymptomatic mice were free of PrP(Sc) infection by Western blot and histological examination. These promising findings suggest that effective mucosal vaccination is a feasible and useful method for overcoming tolerance to PrP and preventing prion infection via an oral route. PMID- 18407425 TI - Cognitive and neuroinflammatory consequences of mild repeated stress are exacerbated in aged mice. AB - Peripheral immune stimulation as well as certain types of psychological stress increases brain levels of inflammatory cytokines such as interleukin-1beta (IL 1beta), IL-6 and tumor necrosis factor alpha (TNFalpha). We have demonstrated that aged mice show greater increases in central inflammatory cytokines, as well as greater cognitive deficits, compared to adults in response to peripheral lipopolysaccharide (LPS) administration. Because aged mice are typically more sensitive to systemic stressors such as LPS, and certain psychological stressors induce physiological responses similar to those that follow LPS, we hypothesized that aged mice would be more sensitive to the physiological and cognitive effects of mild stress than adult mice. Here, adult (3-5 months) and aged (22-23 months) male BALB/c mice were trained in the Morris water maze for 5 days. Mice were then exposed to a mild restraint stress of 30 min before being tested in a working memory version of the water maze over a 3-day period. On day 4 mice were stressed and then killed for collection of blood and brain. In a separate group of animals, mice were killed immediately after one, two or three 30 min restraint sessions and blood was collected for peripheral corticosterone and cytokine protein measurement, and brains were dissected for central cytokine mRNA measurement. Stress disrupted spatial working memory in both adult and aged mice but to a much greater extent in the aged mice. In addition, aged mice showed an increase in stress-induced expression of hippocampal IL-1beta mRNA and MHC class II protein compared to non-stressed controls while expression in adult mice was unaffected by stress. These data show that aged mice are more sensitive to both the cognitive and inflammatory effects of mild stress than are adult mice and suggest a possible role for IL-1beta. PMID- 18407427 TI - A revision of the possible dual role of Helicobacter pylori in myelodysplastic syndrome. PMID- 18407426 TI - Salivary cortisol is associated with diagnosis and severity of late-life generalized anxiety disorder. AB - Age-associated alterations in hypothalamic-pituitary-adrenal (HPA) axis functioning may make individuals more susceptible to HPA dysregulation in the context of mood and anxiety disorders. Little to no research has been done to examine HPA axis function in generalized anxiety disorder (GAD), particularly in late-life GAD, the most prevalent anxiety disorder in the elderly. The study sample consisted of 71 GAD subjects and 40 nonanxious comparison subjects over 60 years of age. We examined the hypotheses that elderly individuals with GAD will have elevated salivary cortisol levels compared to nonanxious subjects, and that elevated cortisol levels in GAD will be associated with measures of symptom severity. We report that late-life GAD is characterized by elevated basal salivary cortisol levels, with higher peak cortisol levels and larger areas under the curve, compared to nonanxious subjects. Additionally, severity of GAD as measured by the GAD Severity Scale and the Penn State Worry Questionnaire are positively correlated with cortisol levels. These data demonstrate HPA axis dysfunction in late-life GAD and suggest the need for additional research on the influence of aging on HPA axis function in mood and anxiety disorders. PMID- 18407428 TI - Is a short-interval postradiation mammogram necessary after conservative surgery and radiation in breast cancer? AB - PURPOSE: To examine, in a retrospective study, whether the initial posttreatment mammogram offers any benefit to patients. METHODS AND MATERIALS: Patients were selected who had radiation after breast-conservation therapy from 1995 through 2005 and had follow-up mammography at the University of California-Los Angeles (UCLA) within 1 year of completing radiotherapy. Results of the initial follow-up mammogram were analyzed to determine the yield of this initial mammogram. RESULTS: Between 1995 and 2005, 408 patients treated with breast-conserving therapy and radiation had follow-up mammograms at UCLA within 1 year of completion of radiation. Median age at radiation completion was 56.9 years. Median interval between radiation and the initial mammogram was 3.1 months. Ten patients were found to have suspicious findings on the initial postradiation mammogram, prompting biopsy, but only 2 were found to have recurrent cancer. None of those lesions were palpable. In both cases the recurrences were ductal carcinoma in situ. Thus, the yield of the initial postoperative mammogram as compared with physical examination findings is estimated at 0.49 recurrences detected per 100 mammograms performed (95% confidence interval 0.059-1.759). CONCLUSIONS: The yield of the initial postradiation mammography at UCLA seems to be low, and only noninvasive carcinomas were found. Our data support the rationale to avoid the initial short-interval postradiation mammography and evaluate patients at 12 months. PMID- 18407429 TI - Combined intensity-modulated radiation therapy and brachytherapy in the treatment of cervical cancer. AB - PURPOSE: This treatment planning study compared pseudo-step-wedge intensity modulation (PSWIM), intensity-modulated radiation therapy (IMRT), and conventional external irradiation, all combined with brachytherapy, for treatment of patients with cervical cancer. METHODS AND MATERIALS: This was a prospective study of 10 patients treated with PSWIM delivering 50.4 Gy to the pelvic lymph nodes and 20 Gy to the cervical tumor. This treatment was compared with a conventional treatment plan with a four-field box to 45 Gy and to an IMRT plan delivering 45 Gy. In each case, brachytherapy was prescribed to a total Point A low-dose-rate equivalent dose of 85 Gy. Total doses to Points A, Point P, the bladder point, and the rectal point were calculated. Acute toxicity and treatment response were prospectively recorded. RESULTS: The mean PSWIM total low-dose-rate equivalent dose to Points A and P (97.3 Gy and 65.1 Gy, respectively) was significantly higher, the mean rectal dose was the same, and the mean bladder dose was higher than with IMRT or four-field box. No acute toxicity of greater Grade 2, as defined by the than Radiation Therapy Oncology Group, was experienced. The positron emission tomography-based treatment response compared favorably with our institutional experience. CONCLUSIONS: Use of PSWIM and brachytherapy delivers significantly more dose to the tumor and lymph nodes than do competing techniques. Rectal doses are comparable. Maximum bladder point doses are higher. Toxicity and tumor response are acceptable. PMID- 18407430 TI - Intensity-modulated radiotherapy as primary therapy for prostate cancer: report on acute toxicity after dose escalation with simultaneous integrated boost to intraprostatic lesion. AB - PURPOSE: To report on the acute toxicity of a third escalation level using intensity-modulated radiotherapy for prostate cancer (PCa) and the acute toxicity resulting from delivery of a simultaneous integrated boost (SIB) to an intraprostatic lesion (IPL) detected on magnetic resonance imaging (MRI), with or without spectroscopy. METHODS AND MATERIALS: Between January 2002 and March 2007, we treated 230 patients with intensity-modulated radiotherapy to a third escalation level as primary therapy for prostate cancer. If an IPL (defined by MRI or MRI plus spectroscopy) was present, a SIB was delivered to the IPL. To report on acute toxicity, patients were seen weekly during treatment and 1 and 3 months after treatment. Toxicity was scored using the Radiation Therapy Oncology Group toxicity scale, supplemented by an in-house-developed scoring system. RESULTS: The median dose to the planning target volume was 78 Gy. An IPL was found in 118 patients. The median dose to the MRI-detected IPL and MRI plus spectroscopy-detected IPL was 81 Gy and 82 Gy, respectively. No Grade 3 or 4 acute gastrointestinal toxicity developed. Grade 2 acute gastrointestinal toxicity was present in 26 patients (11%). Grade 3 genitourinary toxicity was present in 15 patients (7%), and 95 patients developed Grade 2 acute genitourinary toxicity (41%). No statistically significant increase was found in Grade 2-3 acute gastrointestinal or genitourinary toxicity after a SIB to an IPL. CONCLUSION: The results of our study have shown that treatment-induced acute toxicity remains low when intensity-modulated radiotherapy to 80 Gy as primary therapy for prostate cancer is used. In addition, a SIB to an IPL did not increase the severity or incidence of acute toxicity. PMID- 18407431 TI - Radiation-inducible caspase-8 gene therapy for malignant brain tumors. AB - PURPOSE: Patients with malignant gliomas have a poor prognosis. To explore a novel and more effective approach for the treatment of patients with malignant gliomas, we designed a strategy that combines caspase-8 (CSP8) gene therapy and radiation treatment (RT). In addition, the specificity of the combined therapy was investigated to decrease the unpleasant effects experienced by the surrounding normal tissue. METHODS AND MATERIALS: We constructed the plasmid pEGR green fluorescence protein that included the radiation-inducible early growth response gene-1 (Egr-1) promoter and evaluated its characteristics. The pEGR-CSP8 was constructed and included the Egr-1 promoter and CSP8 complementary DNA. Assays that evaluated the apoptosis inducibility and cytotoxicity caused by CSP8 gene therapy combined with RT were performed using U251 and U87 glioma cells. The pEGR-CSP8 was transfected into the subcutaneous U251 glioma cells of nude mice by means of in vivo electroporation. The in vivo effects of CSP8 gene therapy combined with RT were evaluated. RESULTS: The Egr-1 promoter yielded a better response with fractionated RT than with single-dose RT. In the assay of apoptosis inducibility and cytotoxicity, pEGR-CSP8 showed response for RT. The pEGR-CSP8 combined with RT is capable of inducing cell death effectively. In mice treated with pEGR-CSP8 and RT, apoptotic cells were detected in pathologic sections, and a significant difference was observed in tumor volumes. CONCLUSIONS: Our results indicate that radiation-inducible gene therapy may have great potential because this can be spatially or temporally controlled by exogenous RT and is safe and specific. PMID- 18407432 TI - A common sense approach to radiotherapy planning of glioblastoma multiforme situated in the temporal lobe. AB - PURPOSE: Irradiation remains the cornerstone of management for glioblastoma multiforme. The Radiation Therapy Oncology Group and European Organization for Research and Treatment of Cancer advocate encompassing the primary tumor plus a 2 cm margin in the high-dose volume. One shortcoming of this approach is the exposure of critical structures to radiation doses that could exceed organ tolerance. We investigated whether the temporal bone (rather than the aforementioned 2-cm radius) would serve as a barrier to tumor spread when regarded as the anterior margin for temporal lobe lesions. We hypothesized that by using the temporal bone as the radiation field margin, toxicity could be reduced without compromising tumor control. METHODS AND MATERIALS: Between 2003 and 2007, 342 patients with newly diagnosed glioblastoma multiforme were treated with surgery and primary irradiation at our institution. Of these 342 patients, 50 had lesions confined to the temporal lobe. The clinical target volume included the primary lesion, the area of edema when present, and a 2-cm margin, except in the direction of the temporal bone. RESULTS: Of the 50 patients, 40 were available for evaluation. At a median follow-up of 12.95 months, 8 patients had not yet shown signs of tumor progression, 24 had local failure, 7 had distant or mixed (local plus distant) failure, and only 1 patient had failure in the infratemporal fossa. CONCLUSIONS: The results of the study have demonstrated an acceptable level of recurrence when the temporal bone, rather than a 2-cm margin, is used as the anterior border of the clinical target volume. The strategy we have proposed achieves tumor control and respects optic tolerance without resorting to complex, expensive approaches such as intensity-modulated radiotherapy. PMID- 18407433 TI - Prognostic value of pathologic complete response after neoadjuvant therapy in locally advanced rectal cancer: long-term analysis of 566 ypCR patients. AB - PURPOSE: In the literature, a favorable prognosis was observed for complete pathologic response after preoperative therapy (ypCR) in patients with locally advanced rectal cancer. The aim of this study is to verify whether ypCR predicts a favorable outcome in a large series of patients. METHODS AND MATERIALS: The Gastro-Intestinal Working Group of the Italian Association of Radiation Oncology collected clinical data for 566 patients with ypCR (ypT0N0) after neoadjuvant therapy. Eligibility criteria included locally advanced rectal cancer with no evidence of metastases at the time of diagnosis, evidence of ypCR after preoperative radiotherapy +/- chemotherapy (CT). RESULTS: Median radiation dose was 50 Gy. A total of 527 patients (93%) received one of 12 different neoadjuvant CT schedules. Sphincter preservation, anteroposterior resection, and endoscopic surgery were performed in 73%, 22%, and 5% of patients, respectively. Adjuvant CT was administered to 22% of patients. Median follow-up was 46.4 months. Locoregional recurrence occurred in 7 patients (1.6%). Distant metastases occurred in 49 patients (8.9%). Overall, 5-year rates of disease-free survival, overall survival, and cancer-specific survival were 85%, 90%, and 94%, respectively. In multivariate analysis, only age and clinical stage statistically correlated with survival outcome. Adjuvant CT was still of borderline significance (worse for adjuvant CT). No relation was found between survival and neoadjuvant CT schedules. CONCLUSION: A ypCR after neoadjuvant therapy identified a favorable group of patients, even in this large series of 566 patients collected in 61 centers. Locoregional recurrence occurred only in 1.6% patients. PMID- 18407434 TI - Preliminary results of electron intraoperative therapy boost and hypofractionated external beam radiotherapy after breast-conserving surgery in premenopausal women. AB - PURPOSE: To report the acute and preliminary data on late toxicity of a pilot study of boost with electron intraoperative therapy followed by hypofractionated external beam radiotherapy (HEBRT) of the whole breast. METHODS AND MATERIALS: Between June 2004 and March 2007, 211 women with a diagnosis of early-stage breast cancer were treated with breast-conserving surgery. During surgery, an electron intraoperative therapy boost of 12 Gy was administered to the tumor bed. Adjuvant local treatment was completed with HEBRT, consisting of a course of 13 daily fractions of 2.85 Gy to the whole breast to a total dose of 37.05 Gy. Acute toxicity of the breast was evaluated at the end of HEBRT and at 1 month of follow up. Late toxicity was recorded at 6 and 12 months of follow-up. RESULTS: We report the data from 204 patients. The maximal acute skin toxicity was observed at the end of HEBRT (182 patients evaluable) with 7 (3.8%) Grade 3, 52 (28.6%) Grade 2, 123 (67.6%) Grade 1, and no Grade 0 or Grade 4 cases. A total of 108 patients were evaluated for late toxicity. The recorded late skin toxicity was Grade 4 in 1 patient (0.9%), Grade 3 in 1 patient, and Grade 2 or less in 106 patients (98.2%). CONCLUSIONS: The results of this study have shown that electron intraoperative therapy followed by HEBRT allows for the delivery of a high dose to the tumor bed and an adequate dose to the whole breast. This treatment is feasible, compliance is high, and the rate of acute toxicity and the preliminary data on chronic toxicity seem acceptable. PMID- 18407435 TI - Combination external beam radiation and brachytherapy boost with androgen suppression for treatment of intermediate-risk prostate cancer: an initial report of CALGB 99809. AB - PURPOSE: Transperineal prostate brachytherapy (TPPB) can be used with external beam radiation therapy (EBRT) to provide a high-dose conformal boost to the prostate. The results of a multicenter Phase II trial assessing safety of combination of EBRT and TPPB boost with androgen suppression (AST) in treatment of intermediate-risk prostate cancer are present here. MATERIALS AND METHODS: Patients had intermediate-risk prostate cancer. Six months of AST was administered. EBRT to the prostate and seminal vesicles was administered to 45Gy followed by TPPB using either (125)I or (103)Pd to deliver an additional 100Gy or 90Gy. Toxicity was graded using the National Cancer Institute CTC version 2 and the Radiation Therapy Oncology Group late radiation morbidity scoring systems. RESULTS: Sixty-three patients were enrolled. Median follow-up was 38 months. Side effects of AST including sexual dysfunction and vasomotor symptoms were commonly observed. Apart from erectile dysfunction, short-term Grade 2 and 3 toxicity was noted in 21% and 7%, primarily genitourinary related. Long-term Grade 2 and 3 toxicities were noted in 13% and 3%. Two patients had Grade 3 dysuria that resolved with longer follow-up. The most common Grade 2 long-term toxicity was urinary frequency (5%). No biochemical or clinical evidence of progression was noted for the entire cohort. CONCLUSIONS: In a cooperative group setting, combination EBRT and TPPB boost with 6 months of AST was generally well tolerated with expected genitourinary and gastrointestinal toxicities. Further follow-up will be required to fully assess long-term toxicity and cancer control. PMID- 18407436 TI - Prevention of age-related spontaneous mammary tumors in outbred rats by late ovariectomy. AB - BACKGROUND: Breast cancer prevention trials have shown that the antiestrogen tamoxifen inhibits development of estrogen receptor (ER)-positive tumors. In Sprague-Dawley rats, removal of ovarian function in young animals can reduce the incidence of spontaneous age-dependent mammary tumors. However, it is not known whether removal of ovaries late in life, before middle age onset, can still prevent mammary tumor development. METHODS: In this study we used Hsd:Sprague Dawley SD (Hsd) rats to determine the effect of late ovariectomy on mammary tumor development. Intact, sham-ovariectomized and ovariectomized rats were followed until 110 weeks of age, or over their life span. In some experiments, palpable tumors were surgically removed upon presentation. RESULTS: Removal of ovaries before middle age onset ( approximately 5-7 months) inhibited development of spontaneous mammary tumors by 95%. Only one mammary tumor was observed in 19 late ovariectomized animals while 47 total tumors developed in 42 non-ovariectomized animals. Tumor incidence was reduced from 73.8 to 5.3% (relative risk=0.05, 95% CI=0.0072-0.354). The frequency of mammary carcinomas in non-ovariectomized virgin female rats was one in eight rats. Spontaneous rat carcinomas expressed ER and other biomarkers, such as cyclin D1. When palpable tumors were removed by surgical excision, tumor multiplicity increased from 0.76 to 1.61 tumors per rat. Surprisingly, ovariectomy increased the 110-week survival rate and maximum life span of Hsd rats. CONCLUSION: Late ovariectomy prevents spontaneous mammary tumor development in Hsd rats. This animal model may be useful for evaluating novel interventions in breast cancer prevention. PMID- 18407437 TI - [The diagnostic problem of uterine sarcomas: report on a case of carcinosarcoma mimicking a gastro-intestinal disease]. AB - Uterine sarcomas are rare tumours with poor prognosis. The most common presenting symptoms are vaginal bleeding or pelvic pain. We present a case of uterine carcinosarcoma diagnosed retrospectively after surgical specimen analysis in a 69 year-old patient presenting gastro-intestinal complaints. At the time of diagnosis, the patient was in FIGO, stage IV. PMID- 18407438 TI - [Monitoring of anticoagulant treatment in patients with cancer]. AB - Venous thromboembolic events are frequent in patients with cancer. Patients on anticoagulant therapy are both at increased risk for subsequent thromboembolic and bleeding events. Whatever the therapeutic option (vitamin K antagonists [VKA] or low-molecular-weight heparin [LMWH]) a clinic and biological monitoring is required. International Normalized Ratio (INR) variability is increased in patients with cancer as compared with patients without cancer and should be maintained in the target range. Anti-Xa activity measurement may be used in LMWH treatments, in order to detect a possible accumulation in any patients; platelet count monitoring is recommended. Interpretation is most often difficult because of numerous possible causes of thrombopenia in the patient with cancer. Anticoagulant clinics are developed in order to help practitioners and participate in the monitoring of anticoagulant treatments in those patients. PMID- 18407440 TI - Metabolic syndrome after menopause and the role of hormones. AB - OBJECTIVES: The purpose of this review is to focus on the importance of metabolic syndrome (MBS) and its increased prevalence in postmenopausal (PM) women. Also the role of hormonal therapy in PM women with MBS will be discussed. METHODS: Review of the relevant literature and results from recent clinical trials. RESULTS: MBS may occur in 40% of PM women and is largely determined by overweight status and obesity. Weight gain, particularly an increase in central fat mass increases in PM women, beginning a few years prior to menopause. Hormonal Therapy (HT) in normal PM women, generally decreases abdominal fat, but the effect of transdermal estrogen is preferable to oral therapy in this regard. In women with MBS, oral therapy was found to increase leptin and the leptin/adiponectin ratio, while transdermal therapy showed no changes. HT has been found to improve insulin resistance in PM women, although the data are mixed. In women with MBS, oral therapy was found to worsen parameters of insulin resistance, while transdermal therapy had minimal effects overall. Women with MBS have elevations in several inflammation and coagulation factors. Both oral and transdermal HT reduce inflammation markers except for levels of CRP and MMP-9, which increase with oral therapy, but are unaffected by the transdermal route. Oral estrogen has a small pro-coagulant effect, not observed with transdermal therapy, in both normal PM women and those with MBS. The beneficial effects of HT on lipids occur in PM women with and without MBS, although the changes in the latter are minimal. Blood pressure was not affected by HT in women with MBS. CONCLUSIONS: Weight gain and obesity largely drives the increased prevalence of MBS in PM women. Use of HT is beneficial overall for reducing many of the parameters of MBS. Our own data would suggest that in MBS, transdermal therapy may be preferable to oral therapy, at least in standard doses. PMID- 18407439 TI - Peroxisome proliferator di-isodecyl phthalate has no carcinogenic potential in Fischer 344 rats. AB - Di-isodecyl phthalate (DIDP), a peroxisome proliferator-activated receptor-alpha activator, is widely used as a plasticizer in the manufacture of polyvinyl chloride (PVC), and ultimately in typical vinyl applications, particularly wire, cable and toys, etc. To examine its carcinogenic potential, DIDP was fed to Fischer 344 rats in the diet at doses of 0, 400, 2000 and 8000 ppm for 2 years. Briefly, significant decreases in the overall survival and body weights, and increases in the relative weights of kidneys and liver were noted in both sexes of the highest dose groups. However, no treatment-related neoplastic lesions were observed in the internal organs, including the liver. Unlike di(2-ethylhexyl) phthalate (DEHP), DIDP failed to maintain the catalase-inducing potential between early and late expressions of catalase protein from western blotting, immunohistochemistry and enzyme activity measurements. These results suggest that the non-carcinogenicity of DIDP in F344 rats was due to its limited potential for peroxisomal proliferating activity. PMID- 18407441 TI - Compressibility and compactibility of granules produced by wet and dry granulation. AB - The bulk properties, compactibility and compressibility of granules produced by wet and dry granulation were compared applying a rotary tablet press, three different morphological forms of calcium carbonate and two particle sizes of sorbitol. Granules from both granulation methods possessed acceptable flow properties; however, the ground (Mikhart) and cubic (Scoralite) calcium carbonate demonstrated better die-filling abilities in the tablet press than the scalenhedral calcium carbonate (Sturcal). The wet processed granules showed in general larger compression properties. This was explained as these granules were mechanical stronger and had a higher initial porosity. In some cases, a large particle surface area of calcium carbonate and sorbitol resulted in a small, insignificant improvement of the consolidation characteristics. A correlation between the compression and compaction characteristics was demonstrated. PMID- 18407442 TI - Development and evaluation of new sustained-release floating microspheres. AB - A type of multi-unit floating alginate (Alg) microspheres was prepared by the ionotropic gelation method with calcium carbonate (CaCO(3)) being used as gas forming agent. Attempts were made to enhance the drug encapsulation efficiency and delay the drug release by adding chitosan (Cs) into the gelation medium and by coating with Eudragit, respectively. The gastrointestinal transit of optimized floating sustained-release microspheres was compared with that of the non floating system manufactured from identical material using the technique of gamma scintigraphy in healthy human volunteers. It was found that the drug encapsulation efficiency of Cs-Alg microspheres was much higher than that of the Ca-Alg microspheres, and coating the microspheres with Eudragit RS could extend the drug release significantly. Both uncoating and coating microspheres were able to continuously float over the simulated gastric fluid (SGF) for 24h in vitro. Prolonged gastric-retention time (GRT) of over 5h was achieved in the volunteer for the optimized coating floating microspheres (FM). In contrast, non-floating system (NFM) could be emptied within 2.5h. In the present study, a multi-unit system with excellent floating ability, optimum drug entrapment efficiency and suitable drug release pattern has been developed. PMID- 18407443 TI - L-histidine-based pH-sensitive anticancer drug carrier micelle: reconstitution and brief evaluation of its systemic toxicity. AB - A doxorubicin (DOX)-carrier micellar system consisting of poly(histidine)(5K)-b poly(ethylene glycol)(2K) and poly(l-lactic acid)(3K)-b-PEG(2K)-folate has been developed targeting the early endosomal pH and it have been convincingly proved that intracellular high dose strategy using such micelles is effective in overcoming multidrug resistance (MDR) of cancer cells. Due to the low DOX concentrations in the micelle solution obtained by dialysis and the lack of long term stability of the micelles, stable and lyophilized micelle formulations were the subject of investigation reported here by using excipients of sucrose, PEG or Pluronic. The reconstituted micelle solutions were examined by particle size, pH sensitivity, and cytotoxicity for MDR cells and the results were compared with the non-lyophilized micelles. Among tested excipients, Pluronic F127 (33 wt%) added to the polymer/drug solution prior to dialysis resulted in a reconstituted product stable for a week and presented equivalent benefits as the fresh micelle formulation. The blank micelles did not present any apparent systemic toxicity in mice up to 2400 mg/kg i.v. injection (800 mg/kg day) for 3 days). The brief toxicity of reconstituted DOX loaded micelles was examined by the maximum tolerated dose (MTD), which was approximately 7.5-fold higher than free DOX and guaranteeing further animal toxicity and efficacy study. PMID- 18407444 TI - Using a drug to structure its release matrix and release profile. AB - Silicone elastomers have proven to be useful implantable release matrices for hydrophobic drugs. However, their utility for the release of hydrophilic materials is less well developed and, even with the addition of polar excipients such as poly(ethylene oxide) (PEO), burst release profiles are often observed achieving longer term release is more challenging. We report that linoleic acid, initially used to solubilize polar, cationic nicotine in silicone precursors, additionally acted to change the internal morphology of resulting silicone+PEO elastomers. The unexpected consequence of this change was a change in the distribution of hydrophilic domains of PEO/drug within the silicone and the ability to control the rate of release of the drug in vitro. The relationship between excipients, silicone morphology, and release profile is examined. PMID- 18407445 TI - The effects of an herbal medicine Bu-Wang-San on learning and memory of ovariectomized female rat. AB - ETHNOPHARMACOLOGICAL SIGNIFICANCE: Bu-Wang-San (BWS) is a traditional Chinese herbal medicine for the treatment of learning and memory impairment. The effect of BWS on neuroprotection and how BWS increases CA1 dendritic spine synapse density in menopaused women was investigated in the model of ovariectomized (OVX) rats. MATERIALS AND METHODS: Sixteen OVX rats were divided into two groups, the OVX group and OVX+BWS group. After 3 months, Morris water maze was used to assess spatial acquisition and spatial retention. Swim time, swim distance, swim speed, quadrant time and platform crossing were recorded. The ultrastructure of the pyramidal cell and spine synapse density were examined by transmission electron microscopy (TEM). RESULTS: In the spatial acquisition and spatial retention phase of testing, BWS group functioned significantly better than control group. Ultrastructural observation of the hippocampal CA1 region of OVX group showed swelling of mitochondria, the broken and reduced cristas and even crista dissolution; however, the mitochondria were protected well in BWS group. In addition, BWS significantly increased spine synapse density. CONCLUSIONS: These results suggested that BWS could improve cognitive ability of menopause-induced learning and memory impairment. The positive effect of BWS on rat learning and memory was associated with increase of spinal synapse density and protection of mitochondrial function of the pyramidal cell in hippocampal CA1 region from menopause-induced injury. PMID- 18407446 TI - Rehmannia glutinosa: review of botany, chemistry and pharmacology. AB - Rehmannia glutinosa, a widely used traditional Chinese herb, belongs to the family of Scrophulariaceae, and is taken to nourish Yin and invigorate the kidney in traditional Chinese medicine (TCM) and has a very high medicinal value. In recent decades, a great number of chemical and pharmacological studies have been done on Rehmannia glutinosa. More than 70 compounds including iridoids, saccharides, amino acid, inorganic ions, as well as other trace elements have been found in the herb. Studies show that Rehmannia glutinosa and its active principles possess wide pharmacological actions on the blood system, immune system, endocrine system, cardiovascular system and the nervous system. Currently, the effective monomeric compounds or active parts have been screened for the pharmacological activity of Rehmannia glutinosa and the highest quality scientific data is delivered to support the further application and exploitation for new drug development. PMID- 18407447 TI - (1)H MR spectroscopy of inflammation, infection and ischemia of the brain. AB - Different pathologic patterns in multiple sclerosis (MS) are reflected by alterations of metabolites in (1)H MR spectroscopy of the brain. Elevated choline (Cho), lactate (Lac), lipids and macromolecules are reliable markers for acute demyelination regardless of the clinical entity (also in acute disseminated encephalomyelitis). N-acetyl-aspartate (NAA) is a suitable marker for neuronal integrity. It is reduced in acute MS lesions and in normal appearing white matter, even distant to acute and chronic-lesions. Recovery from reduced NAA levels to subnormal values during remyelination, and varying time courses of NAA in normal appearing white matter during relapsing remitting disease indicate the value of this spectroscopic marker for monitoring activity and recovery. Inositol (Ins) is increased in chronic MS lesions being a marker for astrocytic gliosis. In viral disease, Cho and Ins are always increased, whereas a reduction of NAA mostly reflects an advanced or a detoriated clinical state. In bacterial brain abscesses, numerous amino acids, lipids and Lac can be elevated. In ischemia, especially the Lac/NAA in comparison with perfusion and diffusion weighted imaging seems to be a new measure for areas of metabolic need, and may help to better characterise the penumbra of the stroke and the final infarct size. PMID- 18407448 TI - Analysis of acetylcholine, choline and butyrobetaine in human liver tissues by hydrophilic interaction liquid chromatography-tandem mass spectrometry. AB - The strong polar quaternary ammoniums, acetylcholine (ACh), choline (Ch) and butyrobetaine (BB, (3-carboxypropyl)trimethylammonium), are believed playing important roles in liver metabolism. These metabolites are at low levels and are weakly retained on reversed-phase liquid chromatographic (RP-LC) columns. Several hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC MS/MS) methods have been reported to analyze these compounds from different samples. However, no application to human liver tissues has been published. In this study, HILIC-MS/MS method was developed to simultaneously determine these three metabolites in human liver tissues. They were simply extracted from tissue, separated on a HILIC column, and detected by tandem MS in the mode of multiple reaction monitoring (MRM). Further studies on the recovery and repeatability based on real samples indicated the method was accurate and reliable. This method was successfully applied to measure the levels of ACh, Ch and BB in 61 human liver tissue samples including normal, hepatocellular carcinoma (HCC) and matched non-cancerous liver tissues. By comparison of Ch and ACh contents in 29 HCC with their matched non-cancerous liver tissues, it was found that ACh content increased in 11/29 HCC cases and decreased in 13/29 cases. Furthermore, the ACh/Ch ratio increased in 16/29 HCC cases, while it decreased in 8/29 cases. These results strongly indicated that there exist different patterns of ACh content in cancer tissues among HCC patients, thus highlighting the understanding of ACh and its relevant signal pathways in hepatic carcinogenesis and HCC progression. PMID- 18407450 TI - Penetrating femoropopliteal injury during modern warfare: experience of the Balad Vascular Registry. AB - OBJECTIVE: Wounding patterns, methods of repair, and outcomes from femoropopliteal injury have been documented in recent civilian literature. In Operation Iraqi Freedom, as in past conflicts, these injuries continue to be a therapeutic challenge. Therefore, the objective of the current study is to document the pattern of femoropopliteal injuries, methods of repair, and early outcomes during the current military campaign in Iraq. METHODS: From September 1, 2004, to April 30, 2007, all vascular injuries arriving at the Air Force Theater Hospital (the central echelon III medical facility in Iraq; equivalent to a civilian level I trauma center), Balad Air Base, Iraq were prospectively entered into a registry. From this, injuries involving the lower extremities were reviewed. RESULTS: During the 32-month study period, 9289 battle-related casualties were assessed. Of these, 488 (5.3%) were diagnosed with 513 vascular injuries, and 142 casualties sustained 145 injuries in the femoropopliteal domain. Femoral level injury was present in 100, and popliteal level injury occurred in 45. Injuries consisted of 59 isolated arterial, 11 isolated venous, and 75 combined. Fifty-eight casualties were evacuated from forward locations. Temporary arterial shunts were placed in 43, of which 40 (93%) were patent on arrival at our facility. Our group used shunts for early reperfusion before orthopedic fixation, during mass casualty care, or autogenous vein harvest in 11 cases. Arterial repair was accomplished with autogenous vein in 118 (88%), primary means in nine (7%), or ligation in seven (5%). Venous injury was repaired in 62 (72%). Associated fracture was present in 55 (38%), and nerve injury was noted in 19 (13%). Early limb loss due to femoropopliteal penetrating injury occurred in 10 (6.9%). Early mortality was 3.5% (n = 5). CONCLUSIONS: Femoropopliteal vascular injury remains a significant reality in modern warfare. Femoral injuries appear more prevalent than those in the popliteal region. Early results of in-theater repair are comparable with contemporary civilian reports and are improved from the Vietnam era. Rapid evacuation and damage control maneuvers such as temporary shunting and early fasciotomy assist timely definitive repair and appear effective. PMID- 18407449 TI - The effect of prenatal nicotine on mRNA of central cholinergic markers and hematological parameters in rat fetuses. AB - A number of studies have demonstrated the influence of nicotine on fetal development. This study determined the expression of choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), and high-affinity choline transporter (CHT1) in the forebrain and hindbrain following chronic prenatal nicotine exposure in the rat fetus (maternal rats were subcutaneously injected with nicotine at different gestation periods). We also measured the effect of chronic nicotine exposure on fetal blood pO(2), pCO(2), pH, Na(+) and K(+) concentrations, as well as lactic acid levels. Maternal nicotine exposure during pregnancy was associated with a decrease in fetal pO(2) coupled with a significant increase in pCO(2) and lactic acid as well as restricted fetal growth. Additionally, maternal nicotine administration also reduced ChAT, VAChT, and CHT1 mRNA levels in the fetal brain. Nicotine-induced fetal hypoxic responses and reduced cholinergic marker expression in the brain were more severe when nicotine was started in early gestation. Our results provide new information about the effects of repeated exposure to nicotine in utero on the expression of central ChAT, VAChT, and CHT1 in the rat fetus. These results indicate that repeated hypoxic episodes or/and a direct effect of nicotine on the central cholinergic system during pregnancy may contribute to brain developmental problems in fetal origin. PMID- 18407451 TI - The impact of race and insurance type on the outcome of endovascular abdominal aortic aneurysm (AAA) repair. AB - BACKGROUND: Although mortality and complication rates for abdominal aortic aneurysm (AAA) have declined over the last 20 years, operative complication rates and perioperative mortality are still high, specifically for repair of ruptures. The goal of this study was to determine the influence of insurance type and ethnicity while controlling for the influences of potential confounders on procedure selection and outcome following endovascular AAA repair (EVAR). METHODS: Using the Nationwide Inpatient Sample (NIS) database, we identified patients who underwent EVAR repair of ruptured and elective infrarenal AAA, between 1990 and 2003. Insurance type and ethnicity were analyzed against the primary outcome variables of mortality and major complications. The potential confounders of age, gender, operative location, diabetes, and Deyo index of comorbidities, were controlled. RESULTS: Bivariate analyses demonstrated significant differences between insurance types and ethnicity and mortality and complications. Patients who were self pay had adverse outcomes in comparison to Private insurance. Whites encountered less perioperative mortality and postoperative complications than Blacks and Hispanics. CONCLUSIONS: After controlling for previously identified associative factors for AAA outcome, ethnicity and insurance type does influence EVAR surgical outcome. Subsequent studies that break down emergent repair vs elective surgery and that longitudinally stratify delay in surgery, or time to admission may be useful. PMID- 18407452 TI - Prospective randomized trial of ACUSEAL (Gore-Tex) vs Finesse (Hemashield) patching during carotid endarterectomy: long-term outcome. AB - BACKGROUND: Several studies have reported that carotid endarterectomy with patch angioplasty is superior to primary closure. Conventional polytetrafluoroethylene (Gore-Tex, W. L. Gore & Associates, Flagstaff, Ariz) patching has been shown to have results similar to autogenous saphenous vein patching; however, it requires a longer hemostasis time. This study examined the long-term clinical outcome and incidence of restenosis after carotid endarterectomy using the new ACUSEAL (Gore Tex) patching vs Hemashield Finesse (Boston Scientific Corp, Natick, Mass) patching. METHODS: The study randomized 200 patients (1:1) undergoing carotid endarterectomy to 100 with ACUSEAL patching and 100 with Hemashield-Finesse patching. All patients underwent immediate and 1-month postoperative duplex ultrasound studies, which were repeated at 6-month intervals. Kaplan-Meier analysis was used to estimate the freedom from stroke, stroke-free survival, and the risk of restenosis for both groups. RESULTS: The demographic and clinical characteristics, the mean operative diameter of the internal carotid artery, and the length of the arteriotomy were similar in both groups. The mean hemostasis time was 5.1 for the ACUSEAL patching vs 3.7 minutes for Finesse patching (P = .01); however, the mean operative times were similar for both groups (P = .61). The incidence of ipsilateral stroke was 2% for ACUSEAL patching (both early perioperative strokes) vs 3% for Finesse patching (2 early and 1 late stroke) at a mean follow-up of 21 months. The respective cumulative stroke-free rates at 1, 2, and 3 years were 98%, 98%, and 98% for ACUSEAL patching vs 97%, 97%, and 97% for Finesse patching (P = .7). The respective cumulative stroke-free survival rates at 1, 2, and 3 years were 97%, 92%, and 88% for ACUSEAL patching vs 96%, 96%, and 91% for Finesse patching (P = .6). The respective freedom from > or =70% carotid restenosis at 1, 2, and 3 years was 98%, 96%, and 89% for ACUSEAL patching vs 92%, 85%, and 79% for Finesse patching (P = .04). CONCLUSIONS: Carotid endarterectomy with ACUSEAL patching and Finesse patching had similar stroke-free rates and stroke-free survival rates. The mean hemostasis time for the ACUSEAL patch was 1.4 minutes longer than that for the Finesse patch; however, the Finesse patch had higher restenosis rates than the ACUSEAL patch. PMID- 18407453 TI - The effect of metabolic syndrome components on exercise performance in patients with intermittent claudication. AB - PURPOSE: To determine the effect of metabolic syndrome components on intermittent claudication, physical function, health-related quality of life, and peripheral circulation in patients with peripheral arterial disease (PAD), and to identify the metabolic syndrome components most predictive of each outcome measure. METHODS: Patients limited by intermittent claudication with three (n = 48), four (n = 45), or five (n = 40) components of metabolic syndrome were studied. Patients were assessed on PAD-specific measures consisting of ankle-brachial index (ABI), initial claudication distance, absolute claudication distance, physical function measures, health-related quality of life, and calf blood flow and transcutaneous oxygen tension responses after 3 minutes of vascular occlusion. RESULTS: Initial claudication distance (mean +/- SD) progressively declined (P = .019) in those with three (203 +/- 167 m), four (124 +/- 77 m), and five (78 +/- 57 m) metabolic syndrome components, and absolute claudication distance progressively declined (P = .036) in these groups as well (414 +/- 224 m vs 323 +/- 153 m vs 249 +/- 152 m, respectively). Furthermore, compared with patients with only three components of metabolic syndrome, those with all five components had impaired values (P < .05) for peak oxygen uptake, ischemic window, 6-minute walk distance, self-perceived walking ability and health, daily physical activity, health-related quality of life on six of eight domains, calf hyperemia, and calf ischemia after vascular occlusion. Abdominal obesity was the predictor (P < .05) of exercise performance during the treadmill and 6-minute walk tests, as well as physical activity. Elevated fasting glucose was the predictor (P < .05) of peripheral vascular measures, self-perceived walking ability and health, and health-related quality of life. CONCLUSION: PAD patients with more metabolic syndrome components have worsened intermittent claudication, physical function, health-related quality of life, and peripheral circulation. Abdominal obesity and elevated fasting glucose are the metabolic syndrome components that are most predictive of these outcome measures. Aggressively treating these metabolic syndrome components may be particularly important in managing symptoms and long term prognosis of PAD patients. PMID- 18407454 TI - Lymph nodes in the tail of the breast can be missed in standard axillary dissection. AB - AIMS: To determine whether excision of the tail of the breast usually by mastectomy or occasionally wide excision together with formal level 1 axillary node dissection (AND) for early breast cancer influences the quantity of harvested lymph nodes and the detection of axillary metastases. METHODS: Multiple regression and binary logistic regression analysis were performed on lymph node harvest data for level 1 AND performed prior to the adoption of sentinel node biopsy during a five year period from 1997 to 2001 at the Leicestershire Breast Unit, comparing AND with and without excision of the tail of the breast. RESULTS: One thousand six hundred and forty-eight level 1 ANDs were performed with a median node harvest of 14 (3-44). Multiple regression analysis identified that the total node harvest was increased by 1.03 nodes if the tail of the breast was excised (p<0.001) and this was independent of the effect on node count of node positivity. Operating surgeon and reporting pathologist did not influence node count. CONCLUSIONS: The results of this study indicate that low axillary nodes may be missed by AND without excision of the tail of the breast and support the use of targeted sentinel node biopsy that should identify an involved node at any site. PMID- 18407455 TI - [Envenoming by exotic snake in France: risks or threatens?]. PMID- 18407456 TI - [Mamba envenomation in mainland France: management of exotic envenomations needs rethinking]. AB - We report a case of accidental green mamba (Dendroaspis viridis) envenomation in the suburbs of Paris. Although moderate, neurotoxic symptoms were clearly present. Immunotherapy with polyvalent serum FAV-Afrique was decided, but logistical problems prevented reasonably quick serum delivery to the hospital. Despite a spontaneously favourable outcome probably due to minimal envenomation, this case exemplifies a near-miss of the care system. Given the increasing incidence of potentially life-threatening exotic envenomations, management of similar cases should be improved. Besides breeders and health care professionals' information, we suggest that victims of an exotic envenomation be referred to a facility with experienced staff and ready access to a limited bank of carefully chosen antivenins. PMID- 18407457 TI - Current approaches in the treatment of Alzheimer's disease. AB - The management of Alzheimer's disease (AD) has been a long-standing challenge and area of interest. Advances in knowledge of the pathogenesis of disease and an increase in disease burden have prompted investigation into innovative therapeutics over the last two decades. This article reviews the various treatments of AD including those targeted towards cholinergic deficiency, oxidative stress, the amyloid cascade, inflammation, and excitotoxicity. Second generation cholinesterase inhibitors remain the preferred therapy for early and intermediate AD while the glutamate antagonist, memantine, is also approved for advanced stages of disease. Antioxidants may delay disease progression, while data on other experimental therapies remain equivocal at best. Gene therapy directed at neurotropins is currently under investigation with some intriguing preliminary results; however, the number of patients examined is too few to be conclusive. Drugs directly targeting amyloid-beta, particularly the amyloid-beta vaccine, continue to be investigated and their forthcoming results are eagerly anticipated. PMID- 18407458 TI - Eliminating bacteria backbone of naked DNA enhanced hFIX expression and reduced inflammatory response in mice. AB - To test the hypothesis that the persistent high level of transgene expression of linear DNA eliminating bacterial backbone (LDEBB) results from less cytokine induction in vivo. We systematically investigated the effect of circular DNA (C DNA), linear DNA (L DNA) and LDEBB on gene expression in mice by hydrodynamics based plasmid administration, and then determined serum cytokine levels in mice by enzyme linked immunosorbent assay (ELISA). The expression of human clotting factor IX (hFIX) gene in mice treated with LDEBB, L DNA or C DNA reached a maximum 1-day after injection (9809, 6447, 2368 ng/mL), respectively. Thirty days after injection, hFIX concentrations dropped to baseline in mice treated with C DNA group, while L DNA group and LDEBB group decreased to 207 and 377 ng/mL, respectively, at the same time-point. Mice receiving LDEBB encoding hFIX expressed approximately 1.5 to 20-fold more serum hFIX than mice injected with L DNA and C DNA for a period of 8 months, respectively. However, mice receiving LDEBB are much less inflammatory than L DNA and C DNA as shown by a 4-fold reduction in serum levels of both TNF-alpha and IL-12. These results demonstrate that LDEBB is not silenced and is capable of expressing persistently high levels of transgene in vivo, which result from less cytokine induction in vivo. LDEBB provides a promising approach and useful therapeutic strategy to improve naked DNA delivery. PMID- 18407459 TI - Perceived anxiety control as a mediator of the relationship between family stability and adjustment. AB - Previous research has demonstrated a relationship between the constructs of family stability and adjustment. The present study evaluates perceived anxiety control as a mechanism through which association between family stability and adjustment may operate. It was hypothesized that perceived anxiety control would mediate the relationships between family stability and depression and anxiety. Participants completed the following measures: Stability of Activities in the Family Environment (SAFE), the Anxiety Control Questionnaire (ACQ), the Beck Depression Inventory II (BDI-II), and the Beck Anxiety Inventory (BAI). Results of multiple regression analyses largely support a model in which perceived anxiety control mediates the relationship between family stability and both depression and anxiety. Potential ways in which family stability may assist in development of perceived control are discussed. PMID- 18407460 TI - Nuclear organization and morphology of cholinergic, putative catecholaminergic and serotonergic neurons in the brains of two species of African mole-rat. AB - The distribution, morphology and nuclear subdivisions of the cholinergic, putative catecholaminergic and serotonergic systems within the brains of two species of African mole-rat (Cape dune mole-rat -Bathyergus suillus; highveld mole-rat -Cryptomys hottentotuspretoriae) were identified following immunohistochemistry for acetylcholinesterase, tyrosine hydroxylase and serotonin. The aim of the present study was to investigate possible differences in the complement of nuclear subdivisions of these systems by comparing those of the mole-rats to published studies of other rodents. The mole-rats used exhibit a major reduction of the visual system and live a subterranean lifestyle. These wild caught animals also have differing social systems, the Cape dune mole-rat is strictly solitary whereas the highveld mole-rat occurs in social familial units. While these differences, especially that of phenotype, may lead to the prediction of significant differences in the nuclear complement of these systems, we found that all nuclei identified in all three systems in the laboratory rat and other rodents had direct homologs in the brains of the mole-rats studied. There were no additional nuclei in the brains of the mole-rats that are not found in the laboratory rat or other rodents and vice versa. The mole-rats are phylogenetically distant from the laboratory rat, but are still part of the order Rodentia. We conclude that changes in the nuclear organization of the systems studied appear to demonstrate a form of constraint related to the phylogenetic level of the order. PMID- 18407461 TI - Introduction to the ILSI Research Foundation/Risk Science Institute reports from the expert working group on neurodevelopmental endpoints. AB - In 2004 the International Life Sciences Institute (ILSI) Risk Science Institute established an expert working group to assess the lessons learned from the implementation of standardized tests for developmental neurotoxicity in experimental animals. This introduction summarizes the working group process and the four reports from the expert working group addressing: the use of positive controls, understanding variability, appropriate statistical techniques, and interpretation. The reports address the 1991 US Environmental Protection Agency standardized protocol for evaluation of developmental neurotoxicity (DNT) and the 2007 Organisation for Economic Co-operation and Development (OECD) Test Guidelines for DNT. The EPA protocol is comprised of tests for evidence of deficits in neurobehavioral function, including auditory startle habituation, motor activity, associative learning and memory, and neuropathologic examination, including simple morphometric analysis. PMID- 18407462 TI - Anti-apoptotic action of Wnt5a in dermal fibroblasts is mediated by the PKA signaling pathways. AB - Wnts are secreted glycoproteins that control diverse biological processes, such as proliferation, differentiation, and apoptosis. We here found that Wnt5a inhibited apoptosis induced by serum deprivation in primary-cultured human dermal fibroblasts. Anti-apoptotic activity of Wnt5a was not inhibited by a dickkopf-1 (DKK), which blocks the canonical Wnt pathway. On the other hand, loss of function of protein kinase A (PKA), induced by treatment with PKA inhibitors, siRNA-mediated knocking down of endogenous PKA catalytic subunits, or enforced expression of dominant-negative PKA inhibited the Wnt5a anti-apoptotic activity, indicating the involvement of PKA in the Wnt5a anti-apoptotic activity. In agreement, phosphorylation levels of a cAMP response element binding protein (CREB), a representative downstream effector of PKA, the activation of which is known to lead to the pro-survival effects, was elevated by Wnt5a. In addition, Wnt5a increased the nuclear beta-catenin level and treatment with imatinib or ionomycin, either of which blocks the beta-catenin pathway, reduced the anti apoptotic activity of Wnt5a, together suggesting the simultaneous involvement of the beta-catenin-mediated pathway in the Wnt5a anti-apoptotic activity. Based on another finding indicating that Wnt5a upregulated PKA-mediated phosphorylation of glycogen synthase kinase-3beta (GSK-3beta) at serine 9 that caused inactivation of GSK-3beta and subsequently resulted in activation of the beta-catenin pathway, we have speculated that the Wnt5a anti-apoptotic activity may be partially mediated by PKA-mediated phosphorylation of GSK-3beta and subsequent activation of the beta-catenin pathway. PMID- 18407463 TI - Galphao/i-stimulated proteosomal degradation of RGS20: a mechanism for temporal integration of Gs and Gi pathways. AB - The G(s) and G(i) pathways interact to control the levels of intracellular cAMP. Although coincident signaling through G(s) and G(i)-coupled receptors can attenuate G(s)-stimulated cAMP levels, it is not known if prior activation of the G(i) pathway can affect signaling by G(s)-coupled receptors. We have found that activated Galpha(o/i) interact with RGS20, a GTPase activating protein for members of the Galpha(omicron/i) family. Interaction between Galpha(o/i) and RGS20 results in decreased cellular levels of RGS20. This decrease was induced by activated Galpha(o) and Galpha(i2) but not by Galpha(q), Galpha(i1) or Galpha(i3.) The Galpha(o/i)-induced decrease in RGS20 can be blocked by proteasomal inhibitors lactacystin or MG132. Activated Galpha(o) stimulates the ubiquitination of RGS20. The serotonin-1A receptor that couples to G(o/i) reduces the levels of RGS20 and this effect is blocked by lactacystin, suggesting that G(o/i) promotes the degradation of RGS20. Expression of RGS20 attenuates the inhibition of beta-adrenergic receptor-induced cAMP levels mediated by the serotonin-1A receptor. Prior activation of the serotonin-1A receptor results in loss of the RGS20-mediated attenuation, and the loss of attenuation is blocked when lactacystin is included during the prior treatment. These observations suggest that G(o/i)-coupled receptors, by stimulating the degradation of RGS20, can regulate how subsequent activation of the G(s) and G(i) pathways controls cellular cAMP levels, thus allowing for signal integration. PMID- 18407464 TI - Betagamma subunits of G(i/o) suppress EGF-induced ERK5 phosphorylation, whereas ERK1/2 phosphorylation is enhanced. AB - Extracellular signal-regulated kinases (ERKs) play important physiological roles in proliferation, differentiation and gene expression. ERK5 is twice the size of ERK1/2, the amino-terminal half contains the kinase domain that shares the homology with ERK1/2 and TEY activation motif, whereas the carboxy-terminal half is unique. In this study, we examined the cross-talk mechanism between G-protein coupled receptors (GPCRs) and receptor tyrosine kinases, focusing on ERK1/2 and 5. The pretreatment of rat pheochromocytoma cells (PC12) with pertussis toxin (PTX) specifically enhanced epidermal growth factor (EGF)-induced ERK5 phosphorylation. In addition, lysophosphatidic acid (LPA) attenuated the EGF induced ERK5 phosphorylation in LPA(1) receptor- and G(i/o)-dependent manners. On the other hand, LPA alone activated ERK1/2 via Gbetagamma subunits and Ras and potentiated EGF-induced ERK1/2 phosphorylation at late time points. These results suggest G(i/o) negatively regulates ERK5, while it positively regulates ERK1/2. LPA did not affect cAMP levels after EGF treatment, and the reagents promoting cAMP production such as forskolin and cholera toxin also attenuated the EGF induced ERK5 phosphorylation, indicating that the inhibitory effect of LPA on ERK5 inhibition via G(i/o) is not due to inhibition of adenylyl cyclase by Galpha(i/o). However, the inhibitory effect of LPA on ERK5 was abolished in PC12 cells stably overexpressing C-terminus of GPCR kinase2 (GRK2), and overexpression of Gbeta(1) and gamma(2) subunits also suppressed ERK5 phosphorylation by EGF. In response to LPA, Gbetagamma subunits interacted with EGF receptor in a time dependent manner. These results strongly suggest that LPA negatively regulates the EGF-induced ERK5 phosphorylation through Gbetagamma subunits. PMID- 18407465 TI - Duration of early maternal separation and prediction of schizotypal symptoms from early adolescence to midlife. AB - Early childhood experiences influence the capacity for healthy social and emotional development. The present study uses longitudinal data to determine whether early maternal separation predicted the subsequent development of schizotypal personality disorder (SPD) symptoms assessed repeatedly from early adolescence over the following 20 years. Within this community sample (N=766), multilevel linear regression analyses revealed the duration of separation from mother in the first 2 years of life predicted elevated SPD symptoms. This relationship was specific to children with mother-reported early angry emotional behavior. These results provide support for the role of early childhood psychosocial risk factors in the development of subsequent schizophrenia spectrum symptoms in emotionally vulnerable children. PMID- 18407466 TI - Lack of blinding in gating studies. PMID- 18407467 TI - Lack of influence of COMT Val158Met genotype on cognition in first-episode non affective psychosis. AB - COMT gene is a logical candidate gene for schizophrenia. Moreover, variations in the COMT Val158Met functional polymorphism have been associated with prefrontal cognitive abnormalities among patients with schizophrenia, healthy relatives and controls. In this study, using an epidemiologically-based sample of 130 patients experiencing a first-episode of a non-affective psychosis, we examined whether COMT Val158Met genotype influenced cognitive performance on the phenotypic expression of psychosis. We found no significant differences in any cognitive measure according to COMT genotype. These findings, together with previously published research, put the relationship between COMT genotype and cognitive performance in doubt. PMID- 18407468 TI - Recent advances in genetic analyses of oculocutaneous albinism types 2 and 4. AB - Patients with OCA are characterized by reduced skin and hair pigmentation and consequent photosensitivity, actinic damage and risk of skin cancer, and by reduced visual acuity and nystagmus. Our survey of Japanese patients revealed that OCA1 was the most frequent type at 34%, while type 2 was present at less than 10%. OCA3 was absent. OCA4, which is a rare type worldwide, was the second most frequent type at 27%. Unexpectedly 10% of the patients turned out to be Hermansky-Pudlak syndrome type 1. Furthermore, the pathogenic p.A481T allele for OCA2, which has 70% melanogenesis activity, was found in approximately 12% of normally pigmented people, indicating that sub-clinical OCA2 might be more frequent in the Japanese than currently thought. And OCA4 is one of the most common types in Japanese patients, despite being rare worldwide. PMID- 18407469 TI - Age-dependent reduction of amygdala volume in bipolar disorder. AB - The amygdala is hypothesized to play a critical role in mood regulation, yet its involvement in bipolar disorder remains unclear. The aim of the present study was to compare measurements of amygdala volumes in a relatively large sample of bipolar disorder patients and healthy controls ranging in age from 18 to 49 years. Subjects comprised 54 adult patients meeting DSM-IV criteria for bipolar disorder and 41 healthy controls matched for age, sex, and education. Magnetic resonance imaging (1.5 T) was performed to obtain volumetric measurements of the amygdala using a manual region-of-interest tracing method with software that allowed simultaneous visualization of the amygdala in three orthogonal planes. The anterior head of the hippocampus was removed in the sagittal plane prior to amygdala volumetry measurement. Multiple regression analysis was computed on amygdala volume measurements as a function of diagnosis, age, sex, and cerebral volume. Bipolar patients showed an age-related reduction of amygdala volume, but controls did not. Among bipolar subjects, amygdala volume was unrelated to medication history. There were no significant hemispheric or sex interactions with the main effects. Results support a role for amygdala dysfunction in bipolar disorder which appears most robustly in older relative to younger adult patients. Differential aging effects in bipolar disorder may compromise amygdala integrity and contribute to mood dysregulation. PMID- 18407470 TI - Light to moderate alcohol use is associated with increased cortical gray matter in middle-aged men: a voxel-based morphometric study. AB - We examined the associations of current alcohol consumption with brain morphometric measures in a healthy, community-dwelling cohort. Cranial T1 weighted 3D-structural MRI scans were obtain in 383 adults (men=211) aged 60-64 years, randomly selected form the larger PATH Through Life study. Voxel-based morphometric analyses were applied to detect regional gray matter and white matter volume changes related to reported weekly alcohol consumption (mean 7.04+/ 8.15 drinks per week). Alcohol consumption in men had a linear association with greater gray matter in bilateral superior and medial frontal gyrus, bilateral middle occipital gyrus, right inferior parietal gyrus, bilateral precentral gyrus, left paracentral gyrus, left uncus and left inferior occipital gyrus, and with lesser white matter in bilateral superior temporal and left parahippocampal gyrus, after adjustment for age, education, total intracranial volume, smoking, hypertension, diabetes and hyperlipidemia. In women, there was no significant linear association between alcohol consumption and total or regional brain volumes. Our results showed a dose-related, sexually dimorphic impact of alcohol on brain tissue volumes independent of cerebrovascular risk factors. These findings are consistent with an inverse-U association between alcohol use and brain morphometry, while suggesting an increased vulnerability of white matter to alcohol-related brain damage. PMID- 18407472 TI - Adsorption of rhodamine B on Rhizopus oryzae: role of functional groups and cell wall components. AB - The role of different functional groups (i.e. amino, carboxyl, hydroxyl as well as phosphate) and cell wall components (such as chitin, chitosan, glucan and phosphomannan) of Rhizopus oryzae on adsorption of rhodamine B is described. The functional groups were chemically modified to determine their contribution in the present adsorption process. Fourier transformed infrared spectroscopic (FTIR) study was used to characterize the modification of the functional groups due to chemical treatments. Carboxyl and amino groups were identified as most important moieties involved in the binding process. Different cell wall components were also isolated from the cell wall to explore their role involved in the binding process. Phosphomannan fraction adsorbed higher amounts of rhodamine B compared to the other cell wall components. Fluorescence microscopic images also supported the differential adsorption capacity of the various cell wall components. PMID- 18407471 TI - Sequential neural changes during motor learning in schizophrenia. AB - Positron emission tomography (PET) was used to investigate differences in neural plasticity associated with learning a unique motor task in patients with schizophrenia and healthy volunteers. Working with a robotic manipulandum, subjects learned reaching movements in a force field. Visual cues were provided to guide the reaching movements. PET rCBF measures were acquired while participants learned the motor skill over successive runs. The groups did not differ in behavioral performance but did differ in their rCBF activity patterns. Healthy volunteers displayed blood flow increases in primary motor cortex and supplementary motor area with motor learning. The patients with schizophrenia displayed an increase in the primary visual cortex with motor learning. Changes in these regions were positively correlated with changes in each group's motor accuracy, respectively. This is the first study to employ a unique arm-reaching motor learning test to assess neural plasticity during multiple phases of motor learning in patients with schizophrenia. The patients may have an inability to rapidly tune motor cortical neural populations to a preferred direction. The visual system, however, appears to be highly compensated in schizophrenia and the inability to rapidly modulate the motor cortex may be substantially corrected by the schizophrenic group's visuomotor adaptations. PMID- 18407473 TI - [What methods can we use to measure inflammation in the asthmatic child?]. AB - Pulmonary function tests remain the best method to assess underlying bronchial inflammation, particularly distal airway flow. However, these tests are limited in their ability to differentiate treatment-reversible small airway obstruction from definitive remodeling related to a viral infection. The increased availability of the exhaled fraction of nitric oxide (FeNO) measurement means that this method is accessible and attractive. In spite of a number of studies, mainly in adults, its use in the management of asthma needs more investigation. Threshold values that permit optimum management are poorly defined, and strategies based on variations of FeNO do not result in reduction of the number of exacerbations, although they do permit better control of drug use and better control of inflammation. Sputum eosinophil counts are of interest for the prevention of exacerbations, although while not well validated they may be useful in some asthmatic children. Also, they require an experienced cytology laboratory. Finally, recent data on bronchial biopsies in children with difficult to-treat asthma will be summarized. PMID- 18407474 TI - [Nutrient intakes in 2005 by non-breast fed French children of less than 36 months]. AB - BACKGROUND: Knowledge of the nutritional consumption of very young children is of main interest, but little is known about the dietary status of French infants and toddlers. OBJECTIVE: To assess energy and nutrient intake and the adequacy of diet of French infants and toddlers. DESIGN AND SETTING: A national cross sectional survey was conducted in France from January to March 2005, using proportionate quota sampling based on the age of the children, the occupation of the mother, and the family socioeconomic category. SUBJECTS: Seven-hundred and six children were allocated to 11 age subgroups ranging from one to three months to 31-36 months. Totally or partially breastfed infants were excluded. ANALYSES PERFORMED: Individual consecutive 3-day weight food records were converted into energy intake and intake of 24 nutrients according to food composition databases recently updated for 1260 standard foods and all the formulae and specific baby foods manufactured and marketed in France in 2005. RESULTS: Mean daily energy intake in 2005 was above the estimated average requirement up to seven months of age, but was lower after one year. Protein, fat, and carbohydrate intakes were adequate, while calcium, magnesium, phosphorus and B group vitamins were above the recommended dietary allowances for all 11 subgroups. However, for toddlers over 12 months of age, some may have had an inadequate intake of alpha-linolenic acid, vitamin E, vitamin C, iron and zinc, whereas mean sodium intake was above the adequate intake for all age subgroups. CONCLUSIONS: The diet of French infants was adequate for a large proportion of children and satisfied most of their nutritional requirements. However, the intake of iron and alpha-linolenic acid in particular needs to be improved for some French toddlers. PMID- 18407475 TI - Cerebellar toxicity with capecitabine in a patient with metastatic breast cancer. PMID- 18407476 TI - Radiotherapy after radical prostatectomy for adenocarcinoma of the prostate: a UK institutional experience and review of published studies. AB - AIMS: The role of radiotherapy to the prostate bed after radical prostatectomy is the subject of much debate. We carried out a retrospective analysis of all patients treated with either adjuvant radiotherapy (ART) or salvage radiotherapy (SRT) in a single UK cancer centre and compared outcomes with published studies. MATERIALS AND METHODS: All patients receiving radiotherapy at any time after a radical prostatectomy were identified and data collected. Patients were referred for ART because of positive surgical margins. SRT was carried out in patients with a detectable or rising prostate-specific antigen (PSA) postoperatively. Patients received either 55 Gy in 20 fractions or 60-64 Gy in 30-32 fractions. All but eight patients were treated using three-dimensional conformal radiotherapy. Both groups were combined for statistical analysis. Biochemical progression-free survival (BPFS) was calculated and displayed using Kaplan-Meier curves. Cox regression was used for univariate and multivariate analysis. RESULTS: In total, 40 patients received postoperative radiotherapy and had a 3 year overall BPFS of 64%. There was no significant difference in 3-year BPFS between ART and SRT (73% vs 61%, P=0.33). Univariate analysis showed that 3-year BPFS was significantly longer if the highest postoperative PSA was<0.5 ng/ml compared with> or =0.5 ng/ml (83% vs 47%, P=0.019), and if the Gleason grade was <7 compared with > or =7 (92% vs 49%, P=0.007). A PSA at diagnosis<10 ng/ml, positive surgical margins, absence of seminal vesicle involvement and neoadjuvant hormones were all associated with a trend towards improved BPFS. Patients with all of these factors had a 3-year BPFS of 91%. Multivariate analysis of the same parameters showed that only Gleason grade remained statistically significant (P=0.019). CONCLUSIONS: The results from this series are in line with published studies, and support the evidence that prostate bed radiotherapy may affect biochemical control in a proportion of patients at risk of relapse. It is not clear whether ART in patients at high risk of relapse or SRT on relapse is most effective. PMID- 18407477 TI - Fitness, fatness, and chronic kidney disease. PMID- 18407478 TI - Cationic lipid/DNA complexes induce TNF-alpha secretion in splenic macrophages. AB - Cationic lipids are widely used as vectors to deliver DNA into mammalian cells in vitro and in vivo. However, cationic lipid/DNA lipoplexes induce an inflammatory response, characterized by pro-inflammatory cytokine secretion, which severely limits their use. The main goal of this work is to identify the organs and the cell type involved in TNF-alpha secretion after lipoplex injection. We determined the kinetics of distribution of the cationic lipid/DNA complex in blood, lung, liver and spleen and quantified the TNF-alpha amount in organ homogenates and in the serum at different points of times. Increase in TNF-alpha production was only observed in the spleen and no significant increase of TNF-alpha production could be observed in the other organs. Fractionation of spleen cells revealed that macrophages were mainly responsible for TNF-alpha secretion. This observation was verified in vivo by using macrophage-removing agents. In conclusion, we show here that the TNF-alpha secreted in the serum after intravenous injection of lipoplexes comes mainly from the splenic macrophages. PMID- 18407479 TI - Role of trehalose in moisture-induced aggregation of bovine serum albumin. AB - Moisture-induced aggregation has been identified as a key problem in the long term storage stability of therapeutic proteins. In the present work, we have investigated the impact of the disaccharide trehalose on the aggregation behavior of a model protein, bovine serum albumin (BSA) under moist conditions. About 50% aggregation of BSA was observed at a moisture level of 8 microl/10mg protein. Including trehalose in the protein sample caused a significant reduction in aggregation. We address the probable mechanisms for the protective effect of trehalose by considering the various hypotheses that have been proposed in the literature. The techniques that have been used include denaturing and non denaturing gel electrophoresis and tryptophan intrinsic fluorescence. The nature of the aggregates was studied by carrying out electrophoresis of the aggregated protein in the presence of reducing and chaotropic reagents. The interaction studies of aggregated BSA with Thioflavin T and CongoRed indicate the possibility of amyloid type of character in the former. These studies may explain the protective role of trehalose under conditions where the storage stability of therapeutic proteins is compromised. PMID- 18407480 TI - Antioxidant effects of methionine, alpha-lipoic acid, N-acetylcysteine and homocysteine on lead-induced oxidative stress to erythrocytes in rats. AB - Lead, widely used in industry, is a great environmental health problem. Many studies have examined its effects on the health of both humans and animals. Experimental studies have shown that sulphur-containing antioxidants have beneficial effects against the detrimental properties of lead. The present study was designed to investigate markers of oxidative stress (hemoglobin (Hb) in whole blood, malondialdehyde (MDA) in sera; superoxidase dismutase (SOD) and glutathione peroxidise (GSH-Px) in erythrocyte hemolysate and vitamins A and E in plasma) in rats given lead (2000ppm) with or without sulphur-containing antioxidants (l-methionine (Met) (100mg/kg/day), N-acetylcysteine (NAC) (800mg/kg/day), l-homocysteine (Hcy) (25mg/kg/day), lipoic acid (LA) (50mg/kg/day)) in their water for 5 weeks. In the lead group, Hb and plasma vitamin E levels were significantly lower whereas MDA levels were significantly higher compared to controls (p<0.05). Hb levels in lead-methionine and lead-LA groups were significantly higher than the lead group (p<0.01). MDA levels were reduced in all groups compared to the lead group (p<0.01). There was a decrease below control values in erythrocyte SOD (p<0.01) and GSH-Px (p<0.05) levels in the lead-LA group. Plasma vitamin A levels were significantly high in lead methionine group compared to lead group (p<0.01). In conclusion, the data suggests that oxidative stress induced by lead is reduced by sulphur-containing compounds. PMID- 18407481 TI - Modifications of interdental papilla microcirculation: a possible cause of periodontal disease in Hashimoto's thyroiditis? AB - BACKGROUND: Hypothyroidism is defined by a decrease in thyroid hormone production and thyroid gland function. The aim of the present research has been to evaluate the morphologic interdental papilla microcirculation of patients suffering from Hashimoto thyroiditis (HT) and to evaluate a possible correlation with the associated periodontal disease. METHODS: Fifteen healthy subjects and 15 patients suffering from HT were examined. The patients who showed conditions known to compromise microcirculation, such as diabetes, hypertension and pharmacological treatments, were not included in the group of healthy patients. All patients were non-smokers. Gingival capillaroscopy was used to investigate the characteristics of microcirculation. Visibility, course, tortuosity, the average caliber of the capillary loops and the number of visible capillary loops per square millimeter were evaluated for each patient. RESULTS: An interdental papilla vascular modification results in HT. In patients suffering from HT, it was possible to observe a reduced caliber of capillaries, as well as a greater number and tortuosity of capillary loops. CONCLUSIONS: This study shows that capillary alterations in patients suffering from HT occur in gingival microcirculation. PMID- 18407482 TI - Indicators from archaeal secretomes. AB - Just as in the Eukarya and the Bacteria, members of the Archaea need to export proteins beyond the cell membrane. This would be required to fulfill a variety of essential functions such as nutrient acquisition and biotransformations, maintenance of extracellular structures and more. Apart from the Eukarya and the Bacteria however, members of the Archaea share a number of unique characteristics. Does this uniqueness extend to the protein secretion system? It was the objective of this study to answer this question. To overcome the limited experimental information on secreted proteins in Archaea, this study was carried out by subjecting the available archaeal genomes, which represent halophiles, thermophiles, and extreme thermophiles, to bioinformatics analysis. Specifically, to examine the properties of the secretomes of the Archaea using the ExProt program. A total of 24 genomes were analyzed. Secretomes were found to fall in the range of 6% of total ORFs (Methanopyrus kandleri) to 19% (Halobacterium sp. NRC-1). Methanosarcina acetivorans has the highest fraction of lipoproteins (at 89) and the lowest (at 1) were members of the Thermoplasma, Pyrobaculum aerophilum, and Nanoarchaeum equitans. Based on the Tat consensus sequence, contribution of these secreted proteins to the secretomes were negligible, making up 8 proteins out of a total of 7105 predicted exported proteins. Amino acid composition, an attribute of signal peptides not used as a selection criteria by ExProt, of predicted archaeal signal peptides show that in the haloarchaea secretomes, the frequency of the amino acid Lys is much lower than that seen in bacterial signal peptides, but is compensated for by a higher frequency of Arg. It also showed that higher frequencies for Thr, Val, and Gly contribute to the hydrophobic character in haloarchaeal signal peptides, unlike bacterial signal peptides in which the hydrophobic character is dominated by Leu and Ile. PMID- 18407483 TI - Determination and pharmacokinetics of gastrodin and p-hydroxybenzylalcohol after oral administration of Gastrodia elata Bl. extract in rats by high-performance liquid chromatography-electrospray ionization mass spectrometric method. AB - In this report, a high-performance liquid chromatography-electrospray ionization mass spectrometric (HPLC-MS) method was used to determine gastrodin (GAS) and p hydroxybenzylalcohol (HBA) in rat plasma after oral administration of Gastrodia elata Bl. (Chinese name: Tianma) extract. Up to 200 microl of plasma containing GAS, HBA and pyromucic acid (as internal standard, IS) were deproteinized with six volumes of methanol. Calibration curves showed linearity within the concentration range tested 2.00-200.00 microg/ml for GAS and 0.832-104.00 microg/ml for HBA in plasma with a correlation coefficient (r) greater than 0.9997. The limit of quantification of 2.00 and 0.83 microg/ml for GAS and HBA had been achieved, respectively. The intra-day and inter-day precisions of the method were determined to be less than 17.82% for GAS and 10.21% for HBA. The recoveries were in the range of 91.12-108.64% with RSD less than 7.80% for GAS and 92.91-106.14% with RSD less than 4.30% for HBA. Evidence showed that a rapid, simple and reproducible LC-MS assay was established to determine GAS and HBA in rat plasma. PMID- 18407484 TI - The use of adjunctive corticosteroids in the treatment of pericardial, pleural and meningeal tuberculosis: do they improve outcome? AB - Tuberculosis remains a major cause of mortality and morbidity on a global scale. Effective anti-tuberculous chemotherapy has improved outcomes for individuals suffering from tuberculosis, although the disease often results in significant and permanent damage to organs. The use of adjunctive corticosteroid treatment has been studied with a view to demonstrating a reduction in inflammatory events that may improve outcomes for both mortality and morbidity. Cochrane reviews have summarized the evidence for adjunctive corticosteroids in the treatment of tuberculous pericarditis, meningitis and pleural effusion. These reviews have shown improved mortality for pericarditis and meningitis, but inconclusive effects for pericardial constriction and ongoing neurological disability. Rapid improvements in clinical parameters for pleural effusion were not supported by any lasting improved outcomes for these patients. PMID- 18407485 TI - Degradation of lignocellulosic materials under sulfidogenic and methanogenic conditions. AB - Biochemical methane potential (BMP) assays, typically used to assess anaerobic biodegradability of liquid wastes with added nutrients and bacteria, were adapted to compare hydrolysis of lignocellulosic material under sulfidogenic and methanogenic environments. A method based on selective inhibition of microorganism activity, by 3% toluene, was used to measure the hydrolysis rate of lignocellulosic material and the accumulation of sugar. The neutral sugars, galactose, glucose, and xylose, which were released from lignocellulosic material such as office paper and newspaper in the presence 3% toluene, clearly accumulated over time under sulfidogenic conditions. The accumulation rates of sugars, glucose, and xylose, were higher in the sulfidogenic condition than in the methanogenic condition, indicating a faster degradation of lignocellulosic materials under the sulfidogenic condition. PMID- 18407486 TI - Towards Q-PCR of pathogenic bacteria with improved electrochemical double-tagged genosensing detection. AB - A very sensitive assay for the rapid detection of pathogenic bacteria based on electrochemical genosensing has been designed. The assay was performed by the PCR specific amplification of the eaeA gene, related with the pathogenic activity of Escherichia coli O157:H7. The efficiency and selectivity of the selected primers were firstly studied by using standard Quantitative PCR (Q-PCR) based on TaqMan fluorescent strategy. The bacteria amplicon was detected by using two different electrochemical genosensing strategies, a highly selective biosensor based on a bulk-modified avidin biocomposite (Av-GEB) and a highly sensitive magneto sensor (m-GEC). The electrochemical detection was achieved in both cases by the enzyme marker HRP. The assay showed to be very sensitive, being able to detect 4.5 ng microl(-1) and 0.45 ng microl(-1) of the original bacterial genome after only 10 cycles of PCR amplification, when the first and the second strategies were used, respectively. Moreover, the electrochemical strategies for the detection of the amplicon showed to be more sensitive compared with Q-PCR strategies based on fluorescent labels such as TaqMan probes. PMID- 18407487 TI - Nanoelectrode ensembles as recognition platform for electrochemical immunosensors. AB - In this study we demonstrate the possibility to prepare highly sensitive nanostructured electrochemical immunosensors by immobilizing biorecognition elements on nanoelectrode ensembles (NEEs) prepared in track-etch polycarbonate membranes. The gold nanodisk electrodes act as electrochemical transducers while the surrounding polycarbonate binds the antibody-based biorecognition layer. The interaction between target protein and antibody is detected by suitable secondary antibodies labelled with a redox enzyme. A redox mediator, added to the sample solution, shuttles electrons from the nanoelectrodes to the biorecognition layer, so generating an electrocatalytic signal. This allows one to fully exploit the highly improved signal-to-background current ratio, typical of NEEs. In particular, the receptor protein HER2 was studied as the target analyte. HER2 detection allows the identification of breast cancer that can be treated with the monoclonal antibody trastuzumab. NEEs were functionalized with trastuzumab which interacts specifically with HER2. The biorecognition process was completed by adding a primary antibody and a secondary antibody labelled with horseradish peroxidase. Hydrogen peroxide was added to modulate the label electroactivity; methylene blue was the redox mediator generating voltammetric signals. NEEs functionalized with trastuzumab were tested to detect small amounts of HER2 in diluted cell lysates and tumour lysates. PMID- 18407488 TI - Telomeres in drag: Dressing as DNA damage to engage telomerase. AB - The telomere field concentrates both on mechanisms of telomere synthesis and the mechanisms by which telomeres protect chromosome termini from fusion and degradation. Recent studies show that the DNA damage response (DDR) machinery, formerly thought to be the culprit in deleterious telomeric fusion and degradation reactions, plays an active role not only in telomere protection but also in regulating telomere synthesis. Conversely, semi-conservative DNA replication, responsible for the bulk of telomere synthesis, now appears to be a pivotal event on the road to telomere de-protection. These advances prompt the notion that the two guises of telomere function are intricately entangled. Indeed, telomeres appear to expose themselves to the DDR upon passage of the replication fork, in turn attracting telomerase. PMID- 18407489 TI - Paediatric palliative care: coming of age in oncology? AB - Palliative care in children has been emerging as a clinical subspecialty of paediatrics for many years. It requires the knowledge and experience of a paediatrician, combined with the skills of a palliative care specialist. Both are essential, as a paediatrician may not have advanced knowledge of palliative care and a palliative care specialist is unlikely to be familiar with the complexity of working with families where the child is the patient. This paper reviews recent literature and discusses advances in the development of palliative care services for children and young people with incurable cancer. It highlights key areas where paediatric palliative care differs from that of adults and outlines the barriers to providing palliation and conducting evidence-based research in children and young people dying from cancer. PMID- 18407490 TI - Steroids and the metabolic syndrome. PMID- 18407491 TI - Synthesis, biochemical evaluation of a range of potent 4-substituted phenyl alkyl imidazole-based inhibitors of the enzyme complex 17alpha-Hydroxylase/17,20-Lyase (P45017alpha). AB - We report the synthesis, biochemical evaluation and rationalisation of the inhibitory activity of a number of azole-based compounds as inhibitors of the two components of the cytochrome P-450 enzyme 17alpha-hydroxylase/17,20-lyase (P450(17alpha)), i.e. 17alpha-hydroxylase (17alpha-OHase) and 17,20-lyase (lyase). The results suggest that the compounds synthesised are potent inhibitors, with 7-phenyl heptyl imidazole (11) (IC(50)=320 nM against 17alpha OHase and IC(50)=100 nM against lyase); 1-[7-(4-fluorophenyl) heptyl] imidazole (14) (IC(50)=170 nM against 17alpha-OHase and IC(50)=57 nM against lyase); 1-[5 (4-bromophenyl) pentyl] imidazole (19) (IC(50)=500 nM against 17alpha-OHase and IC(50)=58 nM against lyase) being the most potent inhibitors within the current study, in comparison to ketoconazole (KTZ) (IC(50)=3.76 microM against 17alpha OHase and IC(50)=1.66 microM against lyase). Furthermore, consideration of the inhibitory activity against the two components shows that all of the compounds tested are less potent towards the 17alpha-OHase in comparison to the lyase component, a desirable property in the development of novel inhibitors of P450(17alpha). From the modelling of these compounds onto the novel substrate heme complex (SHC) for the overall enzyme complex, the length of the compound, along with its ability to undergo interaction with the active site corresponding to the C(3) area of the steroidal backbone, are suggested to play a key role in determining the overall inhibitory activity. PMID- 18407492 TI - Enhanced discrimination and calibration of biomass NIR spectral data using non linear kernel methods. AB - Rapid methods for the characterization of biomass for energy purpose utilization are fundamental. In this work, near infrared spectroscopy is used to measure ash and char content of various types of biomass. Very strong models were developed, independently of the type of biomass, to predict ash and char content by near infrared spectroscopy and multivariate analysis. Several statistical approaches such as principal component analysis (PCA), orthogonal signal correction (OSC) treated PCA and partial least squares (PLS), Kernel PCA and PLS were tested in order to find the best method to deal with near infrared data to classify and predict these biomass characteristics. The model with the highest coefficient of correlation and the lowest RMSEP was obtained with OSC-treated Kernel PLS method. PMID- 18407493 TI - Development of a test method to assess the sludge reduction potential of aquatic organisms in activated sludge. AB - This article shows the development of a quantitative sludge reduction test method, which uses the sludge consuming aquatic worm Lumbriculus variegatus (Oligochaeta, Lumbriculidae). Essential for the test are sufficient oxygen supply and the presence of a non-stirred layer of sludge for burrowing of the organisms. The test eliminates the unwanted effects of the organisms' movements, so-called bioturbation, on oxygen transport and (therefore) on sludge reduction. We used fresh untreated activated sludge grown on sewage, in order to stay close to the daily practice of sludge treatment. By separating sludge and worms, sludge reduction and worm growth are quantified independently and accurately. Predation by L. variegatus approximately doubles the decay rate of activated sludge. A minimum ratio of initial worm to sludge biomass (W0/S0) of about 0.4g worm/g sludge dry mass is required. Under the test conditions 20-40% of the predated sludge is converted into worm biomass. Our test is simple, reproducible and accurate and is done with equipment generally available in any laboratory, yielding results within a few days. The test can also be used to assess the application of mixtures of different aquatic organisms, but does not provide enough information for the design of a sludge treatment reactor. PMID- 18407494 TI - Combined action of a bacterial monooxygenase and a fungal laccase for the biodegradation of mono- and poly-aromatic hydrocarbons. AB - The combined action of a wide substrate range toluene o-xylene monooxygenase from Pseudomonas sp. OX1, able to convert many aromatic compounds into mono- and di hydroxylated derivatives, and fungal laccases from Pleurotus ostreatus which oxidize these hydroxylated products yielding polymers with reduced toxicity is described. This strategy permits to overcome many of the substrate specificity problems and dead end toxic products formation generally encountered in complex bacterial biodegradation pathways. Toluene and naphthalene degradations were tested as representative of mono- and poly-aromatic pollutants. The combined biological action was optimized in micellar and microemulsion systems able to increase the bioavailability of the hydrophobic aromatic pollutants. This approach allows efficient hydroxylations of hydrophobic substrates thus favoring the further action of fungal oxidases. PMID- 18407495 TI - Studies on phosphorus solubilizing activity of a strain of phosphobacteria isolated from chestnut type soil in China. AB - A phosphorus solubilizing bacterium, designated phosphobacterium 9320-SD, was isolated from field soil in Tianjin, China. Cells of the phosphobacterium 9320-SD were gram-positive, rod shaped, and produced spores. When 9320-SD was inoculated into MPMLM, amended with powdered (insoluble) mineral phosphate as the single P source, and incubated at 30 degrees C, the release of soluble phosphorus increased with increasing amounts of added phosphates over the range of 0.12-4% (w/v). The maximal available phosphorus reached 12.01 mmol P/L after 7 days incubation. Furthermore, there was a direct positive correlation (r = 0.9330) between the level of soluble phosphorus release and the concentration of viable bacteria. SEM study of the phosphate powder retrieved from the phosphobacterium 9320-SD cultured medium revealed the actual dissolution of phosphate from the mineral surface. Phosphobacterium 9320-SD had significant effect (p < 0.05) on winter wheat total P and plant biomass under both pot and field conditions, although no obvious difference in plant height was found compared to the control. Taken together, these results demonstrate that phosphobacterium 9320-SD has the ability to convert non-available forms of phosphorus into plant-available forms, and therefore holds great potential for development as a biofertilizer to enhance soil fertility and promote plant growth. PMID- 18407496 TI - Antagonists of the human adenosine A2A receptor. Part 2: Design and synthesis of 4-arylthieno[3,2-d]pyrimidine derivatives. AB - We describe herein the discovery and development of a series of 4-arylthieno[3,2 d]pyrimidines which are potent adenosine A(2A) receptor antagonists. These novel compounds show high degrees of selectivity against the human A(1), A(2B) and A(3) receptor sub-types. Moreover, a number of these compounds show promising activity in vivo, suggesting potential utility in the treatment of Parkinson's disease. PMID- 18407497 TI - Comparison of the effects of octreotide and melatonin in preventing nerve injury in rats with experimental spinal cord injury. AB - In this study, we aimed to investigate the biochemical and histopathological protective effects of octreotide and melatonin in an experimental model of spinal cord injury. Fifty- six male albino Wistar rats were divided into four groups. Rats in the G1 group (n=7; control group) did not undergo any treatment except for anesthesia prior to being killed. Rats in the G2 group (n=7) underwent laminectomy and aneurysmal clip application at the T4-5 level. G3 group rats (n=14) were either treated with a 7.5 mg/kg intraperitoneal dose of melatonin (Sigma, St. Louis, MO, USA) immediately after laminectomy, then the same dose again on the day following injury (G3a), or given three equal doses over 10 days to achieve a total dose of 7.5 mg/kg/day (G3b). G4 group rats (n=14) were either treated with a 30microg/kg intraperitoneal dose of octreotide (Sandostatin; Novartis, Istanbul, Turkey) immediately after laminectomy, then the same dose again on the day following injury (G4a), or given three equal doses over 10 days to achieve a total dose of 30miocrog/kg/day (G4b). Rats in the G3 and G4 groups were sacrificed on days 1 and 10 after spinal cord injury (n=7 at each time point) and spinal cord samples were obtained. Tissue malonyldialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) levels were assayed. G3a, G3b and G4b had significantly lower levels of MDA than G2 (p<0.01). G3b had significantly higher SOD and GSH-Px levels than G2 (p<0.01). Histopathologically, melatonin significantly reduced necrosis and degeneration in both the initial and late stages (p<0.01). Octreotide had significant effects on necrosis and degeneration during the late stages, and on edema and congestion in both the initial and the late stages of injury (p<0.01). Melatonin was found to be superior to octreotide with respect to the prevention of congestion, edema, axonal degeneration and necrosis. PMID- 18407498 TI - Solitary fibrous tumour extending both supratentorially and infratentorially. AB - Solitary fibrous tumours (SFT) of the central nervous system are rare. Such lesions are mesenchymal neoplasms that resemble meningioma. To date, 73 cases of SFT have been reported in the literature, but there is no reported case of an SFT that extends into both the supratentorial and infratentorial spaces. A 76-year old man presented with headache, dysarthria and ataxia of 2 months' duration. MRI revealed a right cerebellar tumour, extending superiorly to the occipital lobe. During surgery, a huge, solid and well-capsulated tumour was observed. The tentorium cerebelli was also damaged by the tumour. Histological and immunohistochemical studies confirmed the diagnosis of SFT. This is the first reported case of SFT located in both the infratentorial and supratentorial spaces. SFT are spindle cell neoplasms with a characteristic immunohistochemical profile of CD34, vimentin and bcl-2 positivity. PMID- 18407499 TI - Three-dimensional reconstructed CT scanning for targeting hypertensive putaminal hemorrhage. AB - We developed an accurate frontal targeting method for hypertensive putaminal hemorrhage (HPH) using three-dimensional (3D) reconstructed CT scanning. Five consecutive unconscious patients with a significant volume of HPH were treated neuroendoscopically via frontal trephination. CT images were examined and reconstructed by an independent neuroradiologist for measuring the selected frontal entry point, depth of penetrating path, and surgical trajectory on the basis of the plane formed by the bilateral orbitomental lines delineated on the reconstructed 3D CT images. All patients regained consciousness post-operatively without complications. All had an accurate trajectory of penetrating path and the average percentage hematoma evacuation was 84%. Use of the 3D reconstructed CT imaging technique combined with neuroendoscopy may prove valuable as a minimally invasive and time-saving method of targeting HPH. Using this method, no further CT scanning is needed for subsequent surgical planning. PMID- 18407500 TI - Ependymal cyst in the lumbar spine associated with cauda equina compression. AB - There are many reports of extradural ependymal cysts in the literature; however, reports of intradural ependymal cysts are very rare and there has been no prior mention of an ependymal cyst originating from the filum terminale. In this report we present the case of a 31-year-old woman with an ependymal cyst that caused cauda equina compression, and discuss the clinical profile of the case in terms of symptoms, diagnostic images, pathohistological findings, and surgical procedures. To our knowledge, this is the first report of an ependymal cyst that caused cauda equina compression. The cyst was successfully treated by excision of the cyst during careful intraoperative monitoring to prevent neurological damage to the conus medullaris and cauda equina. PMID- 18407501 TI - Neuroprotective and antiamnesic effect of donepezil, a nicotinic acetylcholine receptor activator, on rats with concussive mild traumatic brain injury. AB - In this study we evaluated the effect of donepezil on the neurodegeneration and behavioral impairments induced by mild traumatic brain injury (MTBI). Donepezil is an acetylcholinesterase inhibitor that is used to treat Alzheimer's disease. Donepezil was given orally to rats subjected to MTBI. Treatment with a single oral dose of donepezil (12mg/kg) immediately after injury significantly attenuated MTBI-induced neuronal death and cognitive impairment as measured by preservation of neurons in the CA1 region of the hippocampus and a water maze test respectively. However, these neuroprotective effects were prevented by concomitant injection of mecamylamine, a nicotinic acetylcholine-receptor (nAChR) antagonist, indicating that protection is mediated by nAChR activation. PMID- 18407503 TI - Disorders of the suprapatellar pouch of the knee. AB - Proper recognition and treatment of pathological conditions of the suprapatellar pouch of the knee is dependent on the knowledge of normal pouch anatomy and of the various conditions which affect this area of the knee and contribute to knee pain. This article includes a comprehensive review of the surgical anatomy of the pouch, current surgical techniques and review of the common conditions that have a predilection for this often overlooked area of the knee. PMID- 18407504 TI - Does the brake response time of the right leg change after left total knee arthroplasty? A prospective study. AB - Patients undergoing total knee arthroplasty often ask when they can safely resume car driving. There is little evidence available on which physicians can rely when advising patients on this issue. In a prospective study we assessed the brake response time of 24 patients admitted to the clinic for left total knee arthroplasty preoperatively and then 10 days after surgery. On each measurement day the patients performed two tasks, a simple and a complex brake response time task in a car simulator. Ten days after left TKA the brake response time for the simple task had decreased by 3.6% (p=0.24), the reaction time by 3.1% (p=0.34) and the movement time by 6.6% (p=0.07). However, the performance improvement was not statistically significant. Task complexity increased brake response time at both time points. A 5.8% increase was significant (p=0.01) at 10 days after surgery. Based on our results, we suggest that patients who have undergone left total knee arthroplasty may resume car driving 10 days after surgery as long as they drive a car with automatic transmission. PMID- 18407502 TI - Lesion processing: high-fidelity versus lesion-bypass DNA polymerases. AB - When a high-fidelity DNA polymerase encounters certain DNA-damage sites, its progress can be stalled and one or more lesion-bypass polymerases are recruited to transit the lesion. Here, we consider two representative types of lesions: (i) 7,8-dihydro-8-oxoguanine (8-oxoG), a small, highly prevalent lesion caused by oxidative damage; and (ii) bulky lesions derived from the environmental pre carcinogen benzo[a]pyrene, in the high-fidelity DNA polymerase Bacillus fragment (BF) from Bacillus stearothermophilus and in the lesion-bypass DNA polymerase IV (Dpo4) from Sulfolobus solfataricus. The tight fit of the BF polymerase around the nascent base pair contrasts with the more spacious, solvent-exposed active site of Dpo4, and these differences in architecture result in distinctions in their respective functions: one-step versus stepwise polymerase translocation, mutagenic versus accurate bypass of 8-oxoG, and polymerase stalling versus mutagenic bypass at bulky benzo[a]pyrene-derived lesions. PMID- 18407505 TI - Heteroaromatic ester inhibitors of hepatitis A virus 3C proteinase: Evaluation of mode of action. AB - The related 3C and 3C-like proteinase (3C(pro) and 3CL(pro)) of picornaviruses and coronaviruses, respectively, are good drug targets. As part of an effort to generate broad-spectrum inhibitors of these enzymes, we screened a library of inhibitors based on a halopyridinyl ester from a previous study of the severe acute respiratory syndrome (SARS) 3CL proteinase against Hepatitis A virus (HAV) 3C(pro). Three of the compounds, which also had furan rings, inhibited the cleavage activity of HAV 3C(pro) with K(ic)s of 120-240nM. HPLC-based assays revealed that the inhibitors were slowly hydrolyzed by both HAV 3C(pro) and SARS 3CL(pro), confirming the identity of the expected products. Mass spectrometric analyses indicated that this hydrolysis proceeded via an acyl-enzyme intermediate. Modeling studies indicated that the halopyridinyl moiety of the inhibitor fits tightly into the S1-binding pocket, consistent with the lack of tolerance of the inhibitors to modification in this portion of the molecule. These compounds are among the most potent non-peptidic inhibitors reported to date against a 3C(pro). PMID- 18407506 TI - Synthesis and anti-platelet evaluation of 2-benzoylaminobenzoate analogs. AB - Fifty-two 2-benzoylaminobenzoate analogs were synthesized and subjected to anti platelet aggregation assay using arachidonic acid (AA), collagen (Col), thrombin (Thr), and U46619 as inducers. The results revealed that most of 2 benzoylaminobenzoic acid derivatives showed a selectively inhibitory effect on AA induced platelet aggregation. As a result of the 2-benzoylaminobenzoic acid derivatives (18, 44, and 46), there were no inhibitory effects on platelet aggregation induced by U46619, but these elicited an inhibitory effect on thromboxane B(2) formation at 1.0microM. These 2-benzoylaminobenzoate analogs were therefore proposed as cyclooxygenase inhibitors. PMID- 18407507 TI - Characterization of calcium phosphate coatings doped with Mg, deposited by pulsed laser deposition technique using ArF excimer laser. AB - Calcium phosphate layers were deposited on Ti6Al4V substrates with TiN buffer layers by use of pulsed laser deposition method. With this technique three pressed pellets consisted of tricalcium phosphate (TCP, Ca(3)(PO(4))(2)), hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)) and hydroxyapatite-doped with magnesium (HA with 4% of Mg and trace amount of (Ca,Mg)(3)(PO(4))(2)) were ablated using ArF excimer laser (lambda=193 nm). The using of different targets enabled to determine the influence of target composition on the nature of deposited layers. The obtained deposits were characterized by means of Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction method (XRD). The obtained Fourier spectras revealed differences in terms of intensity of spectral bands of different layers. The analysis from XRD showed that Mg-doped HA layer has crystalline structure and TCP and HA layers composition is characterized by amorphous nature. PMID- 18407508 TI - A carrot G-box binding factor-type basic region/leucine zipper factor DcBZ1 is involved in abscisic acid signal transduction in somatic embryogenesis. AB - Carrot (Daucus carota) somatic embryogenesis has been extensively used as an experimental system for studying embryogenesis. In maturing zygotic embryos, abscisic acid (ABA) is involved in acquisition of desiccation tolerance and dormancy. On the other hand, somatic embryos contain low levels of endogenous ABA and show desiccation intolerance and lack dormancy, but tolerance and dormancy can be induced by exogenous application of ABA. In ABA-treated carrot embryos, some ABA-inducible genes are expressed. We isolated the Daucus carota bZIP1 (DcBZ1) gene encoding a G-box binding factor-type basic region/leucine zipper (GBF-type bZIP) factor from carrot somatic embryos. The expression of DcBZ1 was detected in embryogenic cells, non-embryogenic cells, somatic embryos, developing seeds, seedlings, and true leaves. Notably, higher expression was detected in embryogenic cells, true leaves, and seedlings. The expression of DcBZ1 increased in seedlings and true leaves after ABA treatment, whereas expression was not affected by differences in light conditions. During the development of zygotic and somatic embryos, increased expression of DcBZ1 was commonly detected in the later phase of development. The recombinant DcBZ1 protein showed specific binding activity to the two ABA-responsive element-like motifs (motif X and motif Y) in the promoter region of the carrot ABA-inducible gene according to results from an electrophoretic mobility shift assay. Our findings suggest that the carrot GBF type bZIP factor, DcBZ1, is involved in ABA signal transduction in embryogenesis and other vegetative tissues. PMID- 18407509 TI - Outcome after esophagectomy for cancer of the esophagus and GEJ in patients aged over 75 years. AB - OBJECTIVE: Though the surgical treatment of esophageal cancer is increasingly accepted for elderly people defined as aged over 70 years, less is reported about the results in patients over 75. This study is a single institution retrospective analysis of outcome after esophagectomy for cancer of the esophagus and GEJ in patients aged over 75 years. METHODS: All consecutive patients 76 years old and over undergoing curative esophagectomy for cancer in the period 1991-2006 were analyzed as to comorbidities, outcome and long-term survival. All the data had been prospectively collected in a database. Postoperative mortality risk was assessed by P-POSSUM and O-POSSUM score for in-hospital mortality and by the recently published Steyerberg's score system [Steyerberg EW, Neville BA, Koppert LB, Lemmens VEPP, Tilanus HW, Coebergh JWW, Weeks JC, Earle CC. Surgical mortality in patients with esophageal cancer: development and validation of a simple risk score. J Clin Oncol 2006;24:4277-84.] for 30-day mortality. Five-year survival was compared to the standardized survival in the general population. RESULTS: One hundred and eight patients fulfilling the abovementioned criteria were found (76 males and 32 females, mean age 79.5 years, mean standardized life expectancy: 7.36 years). Among them, 69% had esophageal tumors and 31% GEJ tumors. The predominant histology was adenocarcinoma (74%). Eighty-six (79.6%) presented with one or more major comorbidities or a history of previous major upper-GI surgery, potentially affecting the surgical outcome. All underwent resection with curative intent (R(0) 83.3%, R(1) 12%, R(2) 4.6%). The overall postoperative morbidity rate was 51.9%, pulmonary complications (37%) being the most frequent. Postoperative mortality, mainly due to cardiopulmonary complications, was 7.4%, which was consistent with that predicted by P-POSSUM score (7.2%) and lower than that predicted by O-POSSUM score (15.1%). Thirty-day mortality was 5.5%, being consistent with that predicted by the Steyerberg's score (6.8%). Overall 5-year survival was 35.7%, while R(0) overall survival 42% and cancer specific R(0) survival 51.7%. CONCLUSIONS: Patients 76 years old and over with esophageal or GEJ cancer should not be denied surgery solely on the basis of age. Outcome and long-term results in the selected elderly are not differing from those reported for younger patients. However, despite thorough preoperative assessment being applied in the selection of the candidates for surgery, a practical and reliable individual risk-analysis stratification is still lacking. PMID- 18407510 TI - Outcome after extrapleural pneumonectomy for malignant pleural mesothelioma. AB - BACKGROUND: Malignant pleural mesothelioma is a mainly asbestos-related neoplasm that occurs with increasing frequency and is associated with a poor prognosis. Extrapleural pneumonectomy which was initially performed as a stand-alone treatment in patients with resectable disease is now currently almost uniformly applied as part of a multi-modal approach. Its value and advantage over other therapeutic strategies remain points of discussion. We therefore analysed our experience with extrapleural pneumonectomy in the treatment of malignant pleural mesothelioma. METHODS: We retrospectively reviewed our institutional experience with all consecutive patients undergoing extrapleural pneumonectomy for malignant pleural mesothelioma from 1994 to 2005. Patients were analysed with regard to hospital mortality and morbidity and long-term outcome. RESULTS: Forty-nine patients (10 female/39 male, mean age 58+12 years) underwent extrapleural pneumonectomy during the observation period. Median ICU stay was 1 day, median postoperative length of hospital stay was 13 days. After a mean follow-up of 2573 days, median survival was 376 days (mean 672+121 days, range 9-3384). One-year survival was 53%, 3-year survival 27% and 5-year survival 19%. CONCLUSION: Extrapleural pneumonectomy as part of a multi-modality treatment regimen is a good treatment option for selected patients with malignant pleural mesothelioma. The long-term results of this limited series compare favourably to non-surgical treatment regimens. Larger randomised prospective multi-centre trials are warranted to establish clear guidelines. PMID- 18407511 TI - Induction chemoradiation therapy followed by surgical resection for non-small cell lung cancer (NSCLC) invading the thoracic inlet. AB - OBJECTIVE: The role of induction therapy for non-small cell lung cancer (NSCLC) invading the thoracic inlet is unclear. We reviewed our experience with induction chemoradiation followed by surgical resection for NSCLC invading the thoracic inlet. METHODS: We performed a retrospective review of 44 consecutive patients with NSCLC invading the thoracic inlet, treated with induction chemoradiation (two cycles of cisplatin and etoposide concurrently with 45Gy of radiation) followed by surgical resection between 1996 and 2007. RESULTS: All patients underwent chest wall resection (1-5 ribs, mean 3) with resection of the first rib through an anterior (n=15), a posterior (n=18), or a combined approach (n=11). Lobectomy was performed in 40 cases (90%), pneumonectomy in two (5%), and wedge resection in two (5%). Resection of subclavian vessels or portions of vertebrae was performed in five (11%) and 15 (34%) patients, respectively. Hospital mortality was 5% (n=2). R0-resection was achieved in 39 patients (89%). On pathologic examination, 13 patients (30%) showed complete response (pCR) to induction therapy, and 15 (34%) showed minimal microscopic residual disease (90 99% tumor necrosis). The median follow-up was 2 years (range, 2 month-10 years) with an overall cumulative 5-year survival of 59%. Sixteen patients (36%) developed recurrence, which was local in five cases and distant in 11 patients. The 5-year survival in patients with pCR was 90%; 69% in those with minimal residual disease, and 12% in patients with no relevant response (p=0.0005). CONCLUSIONS: Resection of NSCLC invading the thoracic inlet can be performed safely after induction chemoradiation therapy. The response rate after induction therapy is a strong predictor of survival. PMID- 18407512 TI - Multiple endobronchial carcinomas appearing 3 months after segmentectomy for lung cancer. PMID- 18407513 TI - New aspects of adjuvant therapy in endometrial cancer: current standards and future directions. AB - Endometrial cancer is one of the most common gynaecological cancers in western countries. Most women are diagnosed at an early stage of the disease and can be cured by surgery alone. In patients with poor prognostic factors or an advanced disease, the chance of progression-free survival and overall survival is greatly diminished. Adjuvant chemotherapy is effective for patients with advanced disease. The combination of doxorubicin and cisplatin achieves overall response rates ranging from 34 to 60%, and the addition of paclitaxel seems to improve the outcome of patients with advanced disease, but it induces a significantly higher toxicity. A Gynecologic Oncology Study Group phase-III study is currently exploring the triplet paclitaxel+doxorubicin+cisplatin plus G-CSF vs. the less toxic combination of paclitaxel+carboplatin. Ongoing and planned phase-III trials are evaluating newer combination chemotherapy regimens, a combination of irradiation and chemotherapy and the implementation of targeted therapies with the goal of improving the tumour control rate and quality of life. PMID- 18407514 TI - Immune-like mechanisms in ovulation. AB - Ovulation is the unique biological process by which a mature oocyte (egg) and surrounding somatic cells, the cumulus cell-oocyte complex (COC), are released from the surface of the ovary into the oviduct for transport and fertilization. Ovulation is similar to an inflammatory response: the follicles become hyperemic, produce prostaglandins and synthesize a hyaluronan-rich extracellular matrix. However, this view of ovulation might be too restrictive and needs to be broadened to encompass the innate immune cell surveillance-response system. This hypothesis is being proposed because ovarian granulosa cells and cumulus cells express and respond to innate immune cell-related surveillance proteins (Toll like receptors 2 and 4) and cytokines, such as interleukin 6 (IL-6), during ovulation. PMID- 18407515 TI - Reciprocal regulation of bone and energy metabolism. AB - The finding that fat regulates bone metabolism was viewed as an indication that bone might regulate some aspects of energy metabolism in a feedback loop. The search that started nine years ago for a bone-derived hormone that regulates energy metabolism first took a convoluted path through the identification of a modifier gene. Once this hormone, osteocalcin, was identified, it became clear that bone exerts a profound and complex influence on glucose and fat metabolism. This review highlights the most important salient features of this novel regulation of energy metabolism. PMID- 18407516 TI - Lymphoid tissue inducer cells in adaptive CD4 T cell dependent responses. AB - Lymphoid tissue inducer cells (LTi) are known for their role in embryonic lymphoid tissue development. We have identified LTi in adult tissues including spleen and thymus, and provide evidence here for their presence in the submucosa of the intestine. From these observations it is clear that LTi play multiple roles in the adaptive immune system, notably the ability to recruit and sustain primed CD4 T cells in environments where they provide help to other cells. We speculate that their human equivalents could be potential targets for HIV infection and their destruction explains the pattern of loss of CD4 T cells. PMID- 18407518 TI - Characterization of the cell-mediated cytotoxic responses of isogeneic ginbuna crucian carp induced by oral immunisation with hapten-modified cellular antigens. AB - A better understanding of the immune responses in fish elicited by oral immunisation is of importance for the development of new and effective oral vaccines for cultured fish. In the present study, we characterized specific cell mediated cytotoxic responses in isogeneic ginbuna crucian carp (Carassius auratus langsdorfii) following oral immunisation with cellular antigens. Trinitrophenyl- (TNP) or dinitrophenyl- (DNP) modified syngeneic and allogeneic cells were used for studying the fine specificity and genetic restriction of orally-induced cytotoxic cells. Hapten-specific cytotoxic responses were detected in peripheral blood leukocytes (PBLs) of fish orally immunised with haptenated syngeneic cells. PBLs from orally immunised fish had cytolytic activity for haptenated syngeneic cells, but they showed little reactivity against both haptenated and unmodified allogeneic targets. Similarly, oral immunisation of fish with hapten-modified allogeneic cells did not induce hapten-specific cytotoxic cells which can lyse haptenated syngeneic targets. Although ginbuna crucian carp possess spontaneous cytotoxic cells that are capable of killing mammalian tumour cells, cold target inhibition studies suggested that such spontaneous cytotoxic cells were not involved in the killing of haptenated syngeneic targets. Oral immunisation of fish with haptenated syngeneic cells also induced hapten-specific cytotoxic memory responses. Oral administration of haptenated fixed cells also effectively induced hapten-specific cytotoxic cells in the treated fish. These findings suggest that oral immunisation with antigens can elicit antigen-specific cytotoxic cells that are capable of recognizing antigens in an MHC-restricted manner. In addition, our results provide indirect evidence that fish possess a mechanism for taking up exogenous non-replicating antigens from the alimentary tract and generating antigen-specific cytotoxic cells. PMID- 18407519 TI - Cloning, binding properties, and tissue localization of rainbow trout (Oncorhynchus mykiss) ladderlectin. AB - The present paper describes the primary structure, expression and immunohistochemical localization of rainbow trout ladderlectin (RTLL), a multimeric serum lectin that binds Sepharose and LPS of Aeromonas salmonicida. Two rainbow trout cDNAs (504 and 546bp) and a genomic sequence (2kb) were amplified using ladderlectin-specific primers. The sequences were identified as group VII mannose-binding C-type lectins from predicted amino acid sequences and showed highest identity with the Atlantic salmon mannose-binding lectin. The two cDNA sequences (RTLL-1 and RTLL-2) had 92% identity and encoded 173 and 187 amino acids, respectively. The genomic sequence of RTLL, obtained by PCR, was found to encompass six exons and five introns, with exon 2 encoding 14 amino acids which were exclusive to RTLL-2. The relative expression of both transcripts was highest in the renal kidney, while the intestine, gill and skin exhibited higher relative RTLL-2 expression than RTLL-1. RTLL was immunohistochemically present within cells of the branchial epithelium, hepatic sinusoids, biliary epithelium, renal interstitium, skin, and sub-mucosal granular layer of the intestine. RTLL bound galactan-based Sepharose 6B and Sepharose CL-6B matrices but did not bind unmodified acrylic resin base Toyopearl AF-Epoxy 650M, Toyopearl AF-Amino 650M matrices or N-acetylated Toyopearl AF-Amino 650M acrylic matrices. Two dimensional SDS-PAGE and Western blots of whole plasma and plasma proteins which bound chitin and intact bacteria demonstrated multiple electrophoretic isoforms of RTLL ranging in size from 16 to 18kDa and isoelectric points between pH 4.9 and 6.3. These findings show that RTLL is a group VII C-type lectin with multiple isoforms that bind pathogen-associated molecular patterns such as chitin and microbial surfaces. PMID- 18407520 TI - Cloning, characterization and expression analysis of SIMP (source of immunodominant MHC-associated peptides) in grass carp Ctenopharyngodon idella. AB - SIMP (source of immunodominant MHC-associated peptides) plays a key role in N linked glycosylation with the active site of oligosaccharyltransferase, being the source of MHC-peptides in the MHC I presentation pathway. In the present study, the SIMP gene has been cloned from grass carp Ctenopharyngodon idella by rapid amplification of cDNA ends (RACE). The full length of the cDNA sequence is 4384bp, including a 1117bp 5' UTR (untranslated region), a 2418bp open reading frame, and a 849bp 3' UTR. The deduced amino acids of the grass carp SIMP (gcSIMP) are a highly conserved protein with a STT3 domain and 11 transmembrane regions. The gcSIMP spans over more than 24,212bp in length, containing 16 exons and 15 introns. Most encoding exons, except the first and the 15th, have the same length as those in human and mouse. The gcSIMP promoter contains many putative transcription factor binding sites, such as Oct-1, GCN4, YY1, Sp1, Palpha, TBP, GATA-1, C/EBP beta, and five C/EBP alpha binding sites. The mRNA expression of gcSIMP in different organs was examined by real-time PCR. The gcSIMP was distributed in all the organs examined, with the highest level in brain, followed by the level in the heart, liver, gill, trunk kidney, muscle, head kidney, thymus, and the lowest level in spleen. Furthermore, the recombinant gcSIMP has been constructed successfully and expressed in Escherichia coli by using pQE-40 vector, and the polyclonal antibody for rabbit has been successfully obtained, which was verified to be specific. Identification of gcSIMP will help to explore the function in fish innate immunity. PMID- 18407517 TI - Adenomatous polyposis coli plays a key role, in vivo, in coordinating assembly of the neuronal nicotinic postsynaptic complex. AB - The neuronal nicotinic synapse plays a central role in normal cognitive and autonomic function. Molecular mechanisms that direct the assembly of this synapse remain poorly defined, however. We show here that adenomatous polyposis coli (APC) organizes a multi-molecular complex that is essential for targeting alpha3(*)nAChRs to synapses. APC interaction with microtubule plus-end binding protein EB1 is required for alpha3(*)nAChR surface membrane insertion and stabilization. APC brings together EB1, the key cytoskeletal regulators macrophin and IQGAP1, and 14-3-3 adapter protein at nicotinic synapses. 14-3-3, in turn, links the alpha3-subunit to APC. This multi-molecular APC complex stabilizes the local microtubule and F-actin cytoskeleton and links postsynaptic components to the cytoskeleton--essential functions for controlling the molecular composition and stability of synapses. This work identifies macrophin, IQGAP1 and 14-3-3 as novel nicotinic synapse components and defines a new role for APC as an in vivo coordinator of nicotinic postsynaptic assembly in vertebrate neurons. PMID- 18407521 TI - Initiation of signal transduction of the respiratory burst of prawn haemocytes triggered by CpG oligodeoxynucleotides. AB - Phagocytosis is important in the immune system of the prawn and is believed to be a defence parameter. Previous studies have demonstrated that CpG oligonucleotides enhance the activation of the prophenoloxidase activating system of the prawn through either the G-protein/protein kinase C (PKC) or the cAMP pathway. This study investigated the influence of CpG ODN on the respiratory burst used as the indicator of phagocytic activity and on the initiation of the signal pathway in haemocytes of Macrobrachium rosenbergii. When haemocytes were treated in vitro with 50 microg ml(-1) of ODN2006 for 15 min, the increase of nitroblue tetrazolium (NBT) reduction suggested that the respiratory burst of haemocytes can be enhanced by ODN2006 stimulation. In an attempt to determine which signal transduction pathway is involved in the enhancement effect, haemocytes were separately treated with activators or inhibitors of specific signalling components. The results showed that the NBT reduction of haemocytes increased after treatment with sodium fluoride (a G-protein activator) and decreased after treatment with GDP-beta-S (a G-protein inhibitor). When ODN2006-stimulated haemocytes were treated with GDP-beta-S, the inductive effect was significantly reduced. In haemocytes treated with 8-bromo-cAMP (a PKA activator), the NBT reduction was not significantly different from the control. The addition of phosphodiesterase-inhibiting caffeine, which inhibits the degradation of cAMP, decreased the NBT reduction of ODN2006-stimulated haemocytes; however, the addition of phenol-12-myristate-13-acetate (PMA) significantly increased the NBT reduction. When PMA-stimulated haemocytes were treated with chelerythrine (a PKC inhibitor), the induced NBT reduction was significantly reduced. Furthermore, the study of ODN2006-stimulated haemocytes treated with chelerythrine showed that the enhancement effect of ODN2006 on the NBT reduction was significantly decreased. All results suggest that the enhancement of the respiratory burst of prawn haemocytes is induced by ODN2006 via a PKC-activating signalling pathway, but negatively regulated via the cAMP pathway. PMID- 18407522 TI - Novel parameters of surface EMG in patients with Parkinson's disease and healthy young and old controls. AB - The aim of this study was to evaluate a variety of traditional and novel surface electromyography (SEMG) characteristics of biceps brachii muscle in patients with Parkinson's disease (PD) and compare the results with the healthy old and young control subjects. Furthermore, the aim was to define the optimal biceps brachii loading level that would most likely differentiate patients from controls. The results indicated that such nonlinear SEMG parameters as %Recurrence, %Determinism and SEMG distribution kurtosis, correlation dimension and sample entropy were significantly different between the PD patients and healthy controls. These novel nonlinear parameters, unlike traditional spectral or amplitude parameters, correlated with the Unified Parkinson's Disease Rating Scale (UPDRS) and finger tapping scores. The most significant between group differences were found in the loading condition where no additional weights were applied in isometric elbow flexion. No major difference of SEMG characteristics was detected between old and young control subjects. In conclusion, the novel SEMG parameters can differentiate the patients with PD from healthy control subjects and these parameters may have potential in the assessment of the severity of PD. PMID- 18407523 TI - The phenomenological underpinning of the notion of a minimal core self: a psychological perspective. AB - The paper argues that Zahavi's defence of the self as an experiential dimension, i.e. "identified with the first-person givenness of experiential phenomena", and of the notion of a pre-reflective minimal core self relies on an unwarranted assumption. It is assumed that awareness of the phenomenal mode of experiences of objects, i.e. what the object "feels" like for the experiencer, is comparable with, indeed entails, first-person givenness of experience. In consequence both the arguments concerning the foundational role of the pre-reflective minimal core self and the explanation of the unity and identity of the self through time give rise to intractable problems. PMID- 18407524 TI - Structure-specific statistical mapping of white matter tracts. AB - We present a new model-based framework for the statistical analysis of diffusion imaging data associated with specific white matter tracts. The framework takes advantage of the fact that several of the major white matter tracts are thin sheet-like structures that can be effectively modeled by medial representations. The approach involves segmenting major tracts and fitting them with deformable geometric medial models. The medial representation makes it possible to average and combine tensor-based features along directions locally perpendicular to the tracts, thus reducing data dimensionality and accounting for errors in normalization. The framework enables the analysis of individual white matter structures, and provides a range of possibilities for computing statistics and visualizing differences between cohorts. The framework is demonstrated in a study of white matter differences in pediatric chromosome 22q11.2 deletion syndrome. PMID- 18407525 TI - Robust group analysis using outlier inference. AB - Neuroimaging group studies are typically performed with the assumption that subjects used are randomly drawn from a population of subjects. The population of subjects is assumed to have a distribution of effect sizes associated with it that are Gaussian distributed. However, in practice, group studies can include "outlier" subjects whose effect sizes are completely at odds with the general population for reasons that are not of experimental interest. If ignored, these outliers can dramatically affect the inference results. To solve this problem, we propose a group inference approach which includes inference of outliers using a robust general linear model (GLM) approach. This approach models the errors as being a mixture of two Gaussian distributions, one for the normal population and one for the outliers. Crucially the robust GLM is part of a traditional hierarchical group model which uses GLMs at each level of the hierarchy. This combines the benefits of outlier inference with the benefits of using variance information from lower levels in the hierarchy. A Bayesian inference framework is used to infer on the robust GLM, while using the lower level variance information. The performance of the method is demonstrated on simulated and fMRI data and is compared with iterative reweighted least squares and permutation testing. PMID- 18407526 TI - Phylogenetic relationships, character evolution and biogeography of southern African members of Zygophyllum (Zygophyllaceae) based on three plastid regions. AB - The plastid coding rbcL and non-coding trnLF regions of 53 of 55 southern African Zygophyllum species were sequenced and used to evaluate the phylogenetic relationships within the southern African representatives of the genus. Published sequences of the same gene regions of Australian, Asian and North African Zygophyllum species were included to assess the relationships of the species from these regions to the southern African species. The addition of Z. stapffii from Namibia, found to be conspecific with Z. orbiculatum from Angola, lead to a greatly resolved tree. The molecular results were largely congruent with a recent sectional classification of the southern African species and supported their subdivision into subgenera Agrophyllum and Zygophyllum. Reconstruction of the character evolution of capsule dehiscence, seed attachment and seed mucilage showed that these characters allowed a division of southern African species into the two subgenera but that this could not be applied to species occurring elsewhere. Other morphological characters were found to vary and unique character combinations, rather than unique characters, were found to be of systematic value in sectional delimitation. The study suggests that repeated radiations from the horn of Africa to southern Africa and Asia and back lead to the present distribution of the taxa in the subfamily Zygophylloideae. Although this study supports some of the recent taxonomic changes in the group, the unresolved relationships between the proposed genera Tetraena and Roepera and those retained as Zygophyllum species suggest that changes to the taxonomy may have been premature. PMID- 18407527 TI - Westernized-like-diet-fed rats: effect on glucose homeostasis, lipid profile, and adipocyte hormones and their modulation by rosiglitazone and glimepiride. AB - Wersternized diet, containing high fat diet intake combined with high consumption of softdrinks, is accused with the emerge of modern epidemic obesity and diabesity. Therefore, we aimed to study the effect of this diet combination on the homeostasis of glucose, lipids, and some adipohormones in rats and to simulate the metabolic perturbations induced by the unhealthy Westernized diet intake, leading to the development of type 2 diabetes. To achieve this, we divided male Wistar rats (80-120 g) into two main groups: the first was fed commercial normal fat diet and the second received an in-house-prepared high-fat diet (HFD), combined with fructose in drinking water for a period of 6 weeks, followed by a subdiabetogenic dose of streptozotocin (STZ) (35 mg/kg) to produce frank hyperglycemia. The effect of this diet alone or after 2 weeks of treatment with rosiglitazone or glimepiride on glucose homeostasis, lipid profile, and levels of resistin and leptin was studied. The HFD/fructose/STZ diet elevated fasting plasma glucose, fructosamine, insulin, and homeostasis model assessment (HOMA) index, as well as serum triglycerides (TGs), total cholesterol (TC), and low-density lipoprotein cholesterol, with a decrease in high-density lipoprotein cholesterol. Hepatic TG and TC levels, as well as serum activities of aspartate transaminase (AST), alanine transaminase (ALT), and lactate dehydrogenase (LDH), were increased, suggesting a diet-induced hepatic steatosis, beside the increased levels of serum resistin and leptin. Rosiglitazone corrected the altered parameters measured, except for liver TGs; similarly, glimepiride reinstated the inverted parameters but raised insulin level and, consequently, the HOMA index. These results show that this diet could be used to induce an effect that mimics human type 2 diabetes with its metabolic disturbances and is suitable for screening the antidiabetic agents used for management of this disease. PMID- 18407528 TI - Does the hip powder of Rosa canina (rosehip) reduce pain in osteoarthritis patients?--a meta-analysis of randomized controlled trials. AB - OBJECTIVE: Meta-analysis of randomized controlled trials (RCTs)--of a hip powder of Rosa canina (rosehip) preparation for symptomatic treatment of osteoarthritis (OA), in order to estimate the empirical efficacy as a pain reducing compound. METHOD: RCTs from systematic searches were included if they explicitly stated that OA patients were randomized to either rosehip or placebo. The primary outcome was reduction in pain calculated as effect size (ES), defined as the standardized mean difference (SMD). As secondary analysis the number of responders to therapy was analyzed as Odds Ratios (OR), and expressed as the Number Needed to Treat (NNT). Restricted Maximum Likelihood (REML) methods were applied for the meta-analyses using mixed effects models. RESULTS: The three studies (287 patients and a median trial-duration of 3 months)--all supported by the manufacturer (Hyben-Vital International)--showed a reduction in pain scores by rosehip powder (145 patients) compared to placebo (142 patients): ES of 0.37 [95% confidence interval (CI): 0.13-0.60], P=0.002. Test for homogeneity seemed to support that the efficacy was consistent across trials (I(2)=0%). Thus it seems reasonable to assume that the three studies were measuring the same overall effect. It seemed twice as likely that a patient allocated to rosehip powder would respond to therapy, compared to placebo (OR=2.19; P=0.0009); corresponding to a NNT of six (95% CI: 4-13) patients. CONCLUSIONS: Although based on a sparse amount of data, the results of the present meta-analysis indicate that rosehip powder does reduce pain; accordingly it may be of interest as a nutraceutical, although its efficacy and safety need evaluation and independent replication in a future large-scale/long-term trial. PMID- 18407529 TI - Quantitative MR imaging using "LiveWire" to measure tibiofemoral articular cartilage thickness. AB - OBJECTIVE: To assess the reliability and accuracy of manual and semi-automated segmentation methods for quantifying knee cartilage thickness. This study employed both manual and LiveWire-based semi-automated segmentation methods, ex vivo and in vivo, to measure tibiofemoral (TF) cartilage thickness. METHODS: The articular cartilage of a cadaver knee and a healthy volunteer's knee were segmented manually and with LiveWire from multiple 3T MR images. The cadaver specimen's cartilage thickness was also evaluated with a 3D laser scanner, which was assumed to be the gold standard. Thickness measurements were made within specific cartilage regions. The reliability of each segmentation method was assessed both ex vivo and in vivo, and accuracy was assessed ex vivo by comparing segmentation results to those obtained with laser scanning. RESULTS: The cadaver specimen thickness measurements showed mean coefficients of variation (CVs) of 4.16%, 3.02%, and 1.59%, when evaluated with manual segmentation, LiveWire segmentation, and laser scanning, respectively. The cadaver specimen showed mean absolute errors versus laser scanning of 4.07% and 7.46% for manual and LiveWire segmentation, respectively. In vivo thickness measurements showed mean CVs of 2.71% and 3.65% when segmented manually and with LiveWire, respectively. CONCLUSIONS: Manual segmentation, LiveWire segmentation, and laser scanning are repeatable methods for quantifying knee cartilage thickness; however, the measurements are technique-dependent. Ex vivo, the manual segmentation error was distributed around the laser scanning mean, while LiveWire consistently underestimated laser scanning by 8.9%. Although LiveWire offers repeatability and decreased segmentation time, manual segmentation more closely approximates true cartilage thickness, particularly in cartilage contact regions. PMID- 18407530 TI - Molecular characterization of the diversity of Clostridium chauvoei isolates collected from two bovine slaughterhouses: analysis of cross-contamination. AB - Clostridium chauvoei is the etiologic agent of blackleg, a high mortality rate disease affecting mainly cattle and sheep. Carcasses of animals affected by the disease are the chief source of soil infection and considered as an ever-present threat to livestock health. A study was undertaken to examine the cross contamination of C. chauvoei in two different bovine slaughterhouses using restriction endonuclease analysis (REA) and protein analysis. Samples from various sites of two different bovine slaughterhouses were screened and 34 isolates were identified by conventional techniques and 16S rRNA gene (rrs) sequencing. C. chauvoei were isolated from carcass, soil, and sewage from slaughterhouses examined. The isolates were differentiated using REA and whole cell and excretory protein pattern analysis combined with numerical analysis and cluster formation. The alpha and beta toxins produced by the strains were characterized. Our preliminary results suggest that REA combined with numerical analysis provides additional criteria and characteristic banding patterns for the study of the cross-contamination and characterization of C. chauvoei. The effects of temperature, oxygen tension, and enzymes on C. chauvoei hemolysin activity were also discussed. These microorganisms may be a potential contaminant of carcasses and widespread in soil of abattoir environments. The information of area-specific distribution of C. chauvoei strains and its toxin characteristics may give an efficient program in protecting cattle and other ruminants. PMID- 18407531 TI - A single-step assay for the Gerbich-negative allele of glycophorin C. AB - The Gerbich erythrocyte surface protein, glycophorin C (GYPC), can be used by Plasmodium falciparum to invade erythrocytes. The Melanesian Gerbich-negative antigenic condition (Ge(-)) is frequent in some populations where malaria is endemic, suggesting that it protects against malaria. We have determined as precisely as possible the breakpoint of the chromosomal deletion that causes the Ge(-) condition by comparing the partial GYPC sequence of a Papuan Ge(-/-) homozygous individual with known sequences of GYPC. This localisation has allowed us to develop a robust single-step PCR assay suitable for rapid screening of Ge( ). This method is easier to implement than existing methods, can reliably identify heterozygous individuals, and will considerably aid efforts to study the distribution of Ge(-) and its role in protection against malaria. PMID- 18407532 TI - Cognitive outcome in children with rhombencephalosynapsis. AB - PURPOSE: Rhombencephalosynapsis is a rare congenital cerebellar malformation increasingly recognized by prenatal and neonatal neuroimaging. Cognitive outcome seems to be variable but is not well documented. AIMS AND METHODS: To study neurological, behavioural, and cognitive functions of patients with non-syndromic rhombencephalosynapsis, five patients (three female and two male, mean age at the time of this study 8.9 years, range 4.3-17.3 years) were assessed by neurological examination and several tests of behaviour and cognitive functions. RESULTS: Ataxia was present in all patients, but daily life activities were partly restricted in only one. Other symptoms were muscular hypotonia, abnormal eye movements, and head stereotypies. Three patients had pathological scores on both attention and hyperactivity/impulsivity scales. Only two patients had normal full scale IQ (IQ value of 109 and 114, respectively). Verbal and/or performance IQ were impaired in three. CONCLUSION: In non-syndromic rhombencephalosynapsis the clinical presentation is variable. Attention deficit and hyperactivity disorders are frequent behavioural problems. Cognitive functions are mostly impaired, as mild intellectual impairment without a typical cognitive profile. However, rhombencephalosynapsis is compatible with normal cognitive functions. No definitive correlation between cognitive impairment and additional supratentorial abnormalities could be established. PMID- 18407533 TI - Childhood growth and development associated with need for full-time special education at school age. AB - OBJECTIVE: To explore how growth measurements and attainment of developmental milestones in early childhood reflect the need for full-time special education (SE). METHODS: After stratification in this population-based study, 900 pupils in full-time SE groups (age-range 7-16 years, mean 12 years 8 months) at three levels and 301 pupils in mainstream education (age-range 7-16, mean 12 years 9 months) provided data on height and weight from birth to age 7 years and head circumference to age 1 year. Developmental screening was evaluated from age 1 month to 48 months. Statistical methods included a general linear model (growth measurements), binary logistic regression analysis (odds ratios for growth), and multinomial logistic regression analysis (odds ratios for developmental milestones). RESULTS: At 1 year, a 1 standard deviation score (SDS) decrease in height raised the probability of SE placement by 40%, and a 1 SDS decrease in head size by 28%. In developmental screening, during the first months of life the gross motor milestones, especially head support, differentiated the children at levels 0-3. Thereafter, the fine motor milestones and those related to speech and social skills became more important. CONCLUSION: Children whose growth is mildly impaired, though in the normal range, and who fail to attain certain developmental milestones have an increased probability for SE and thus a need for special attention when toddlers age. Similar to the growth curves, these children seem to have consistent developmental curves (patterns). PMID- 18407534 TI - Anti-inflammatory modulation of chronic airway inflammation in the murine house dust mite model. AB - Asthma affects 300 million people worldwide and continues to be a major cause of morbidity and mortality. Disease relevant animal models of asthma are required for benchmarking of novel therapeutic mechanisms in comparison to established clinical approaches. We demonstrate that chronic exposure of mice to house dust mite (HDM) extract results in allergic airway inflammation, that can be significantly attenuated by therapeutic intervention with phosphodiesterase 4 inhibition and corticosteroid treatment. Female BALB/c mice were administered intranasally with HDM (Dermatophagoides pteronyssinus) extract daily for five weeks, and therapeutic intervention with anti-inflammatory treatment (dexamethasone 1 mg/kg subcutaneous once daily, prednisolone 10mg/kg orally twice daily, fluticasone 3, 10 and 30 microg intranasally twice daily, roflumilast 10 mg/kg orally twice daily and intranasally 10 and 30 microg twice daily) was initiated after three weeks of exposure. Chronic HDM extract exposure resulted in significant airway inflammation, demonstrated by bronchoalveolar lavage cell infiltration and lung tissue inflammatory gene expression by TaqMan low density array. Chronic steroid treatment significantly inhibited these parameters. In addition, roflumilast caused a significant reduction in airway inflammatory cell infiltration. We have demonstrated that chronic HDM-induced allergic inflammation can be significantly ameliorated by steroid treatment, and that phosphodiesterase 4 inhibition modulates inflammatory cell infiltration. Therefore, the murine HDM model may be a useful tool for evaluating new targets for the treatment of asthma. PMID- 18407535 TI - The use of transgenic mice for modeling airways disease. AB - Genetic manipulation in mice often combined with models of human disease has allowed investigators to dissect specific disease pathways. These tools have taught us much about the pathogenesis of chronic obstructive pulmonary disease (COPD). In particular, we have defined a complex inflammatory-protease network leading to emphysema. Translation of these findings to humans requires careful interpretation of the strengths and limitations of the models as well as similarities and differences between mouse and human biology. PMID- 18407536 TI - Elongation factor 1Bgamma (eEF1Bgamma) expression during the molting cycle and cold acclimation in the crayfish Procambarus clarkii. AB - Eukaryotic elongation factor 1Bgamma (eEF1Bgamma) is a subunit of elongation factor 1 (EF1), which regulates the recruitment of amino acyl-tRNAs to the ribosome during protein synthesis in eukaryotes. In addition to structural roles within eEF1, eEF1Bgamma has properties which suggest sensory or regulatory activities. We have cloned eEF1Bgamma from axial abdominal muscle of freshwater crayfish, Procambarus clarkii. The predicted amino acid sequence has 66% identity to Locusta migratoria eEF1Bgamma and 65% identity to Artemia salina eEF1Bgamma. We measured eEF1Bgamma expression by real-time PCR, using the relative quantification method with 18s ribosomal RNA as an internal calibrator. eEF1Bgamma expression was lowest in gill, axial abdominal muscle, and hepatopancreas, and was highest in the antennal gland (5.7-fold above hepatopancreas) and cardiac muscle (7.8-fold above hepatopancreas). In axial abdominal muscle, eEF1Bgamma expression was 4.4-fold higher in premolt and 11.9 higher in postmolt compared to intermolt. In contrast, eEF1Bgamma was decreased or unchanged in epithelial tissues during pre- and postmolt. eEF1Bgamma expression in the hepatopancreas was 3.5-fold higher during intermolt compared to premolt and was unchanged in gill and antennal gland. No significant differences in eEF1Bgamma were found after 1 week of acclimation to 4 degrees C. These results show that eEF1Bgamma is regulated at the mRNA level with tissue-specific differences in expression patterns. PMID- 18407539 TI - A clinical comparison of rigid and inflatable endorectal-coil probes for MRI and 3D MR spectroscopic imaging (MRSI) of the prostate. AB - PURPOSE: To compare the data quality and ease of use of four endorectal-coil probe setups for prostate MRI. MATERIALS AND METHODS: Four endorectal-coil probe setups were compared: 1) air-inflated probe; 2) perfluorocarbon (PFC)-inflated probe; 3) rigid, smaller prototype coil; and 4) rigid, smaller coil designed for biopsying the prostate. Signal-to-noise ratio (SNR), positioning, shimming, MRI motion artifact, and MR spectroscopic imaging (MRSI) spectral quality were assessed. RESULTS: Rigid coils provided approximately 2.5-fold higher SNR than inflatable coils near the peripheral zone midline. The biopsy probe sensitivity decreased dramatically by the apex. The rigid probes, as compared to the inflatable probes, took longer to place (10 +/- 2 vs. 7 +/- 2 minutes, P < 0.0002), tended to be placed too superiorly, required repositioning more often (73% vs. 20%, P < 0.003), and had higher motion artifacts (P < 0.001). Shimming time was least for the PFC-inflated probe (2 +/- 0.5 minutes, P < 0.05). The air inflated probe produced larger linewidths (P < 0.01) and tended to have longer shim times (7 +/- 4 minutes) and poorer spectral quality. CONCLUSION: The inflatable coil is a good clinical choice due to ease of use, good coverage, and low motion artifacts. PFC-inflation is recommended as it provided higher quality data than air-inflation. The rigid, smaller probes have higher SNR and produce less tissue distortion and may be preferred for certain applications. PMID- 18407540 TI - Diffusion-weighted PROPELLER MRI for quantitative assessment of liver tumor necrotic fraction and viable tumor volume in VX2 rabbits. AB - PURPOSE: To test the hypothesis that diffusion-weighted (DW)-PROPELLER (periodically rotated overlapping parallel lines with enhanced reconstruction) MRI provides more accurate liver tumor necrotic fraction (NF) and viable tumor volume (VTV) measurements than conventional DW-SE-EPI (spin echo echo-planar imaging) methods. MATERIALS AND METHODS: Our institutional Animal Care and Use Committee approved all experiments. In six rabbits implanted with 10 VX2 liver tumors, DW-PROPELLER and DW-SE-EPI scans were performed at contiguous axial slice positions covering each tumor volume. Apparent diffusion coefficient maps of each tumor were used to generate spatially resolved tumor viability maps for NF and VTV measurements. We compared NF, whole tumor volume (WTV), and VTV measurements to corresponding reference standard histological measurements based on correlation and concordance coefficients and the Bland-Altman analysis. RESULTS: DW-PROPELLER generally improved image quality with less distortion compared to DW SE-EPI. DW-PROPELLER NF, WTV, and VTV measurements were strongly correlated and satisfactorily concordant with histological measurements. DW-SE-EPI NF measurements were weakly correlated and poorly concordant with histological measurements. Bland-Altman analysis demonstrated that DW-PROPELLER WTV and VTV measurements were less biased from histological measurements than the corresponding DW-SE-EPI measurements. CONCLUSION: DW-PROPELLER MRI can provide spatially resolved liver tumor viability maps for accurate NF and VTV measurements, superior to DW-SE-EPI approaches. DW-PROPELLER measurements may serve as a noninvasive surrogate for pathology, offering the potential for more accurate assessments of therapy response than conventional anatomic size measurements. PMID- 18407541 TI - Real-time fast strain-encoded magnetic resonance imaging to evaluate regional myocardial function at 3.0 Tesla: comparison to conventional tagging. AB - PURPOSE: To compare the utility of the real-time technique fast strain-encoded magnetic resonance imaging (fast-SENC) for the quantification of regional myocardial function to conventional tagged magnetic resonance imaging (MRI). MATERIALS AND METHODS: Healthy volunteers (N = 12) and patients with heart failure (N = 7) were examined using tagged MRI and fast-SENC at 3.0T. Circumferential strain was measured using fast-SENC in six endo- and six subepicardial regions in the basal-, mid-, and apical-septum and the basal-, mid , and apical-lateral wall from the four-chamber view. These measurements were plotted to tagging, in corresponding myocardial segments. RESULTS: Peak systolic strain (Ecc) and early diastolic strain rate (Ecc/second) acquired by fast-SENC correlated closely to tagged MRI (r = 0.90 for Ecc and r = 0.91 for Ecc/second, P < 0.001 for both). Both fast-SENC and tagging identified differences in regional systolic and diastolic function between normal myocardium and dysfunctional segments in patients with heart failure (for fast-SENC: Ecc = -21.7 +/- 2.7 in healthy volunteers vs. -12.8 +/- 4.2 in hypokinetic vs. 0.6 +/- 3.8 in akinetic/dyskinetic segments, P < 0.001 between all; Ecc/second = 104 +/- 20/second in healthy volunteers vs. 37 +/- 9/second in hypokinetic vs. -16 +/- 15/second in akinetic/dyskinetic segments, P < 0.001 between all). Quantitative analysis was more time-consuming for conventional tagging than for fast-SENC (time-spent of 3.8 +/- 0.7 minutes vs. 9.5 +/- 0.7 minutes per patient, P < 0.001). CONCLUSION: Fast-SENC allows the rapid and accurate quantification of regional myocardial function. The information derived from fast-SENC during a single heartbeat seems to be superior or equal to that acquired by conventional tagging during several heart cycles and prolonged breathholds. PMID- 18407542 TI - MRI-compatible manipulator with remote-center-of-motion control. AB - PURPOSE: To develop and assess a needle-guiding manipulator for MRI-guided therapy that allows a physician to freely select the needle insertion path while maintaining remote center of motion (RCM) at the tumor site. MATERIALS AND METHODS: The manipulator consists of a three-degrees-of-freedom (DOF) base stage and passive needle holder with unconstrained two-DOF rotation. The synergistic control keeps the Virtual RCM at the preplanned target using encoder outputs from the needle holder as input to motorize the base stage. RESULTS: The manipulator assists in searching for an optimal needle insertion path which is a complex and time-consuming task in MRI-guided ablation therapy for liver tumors. The assessment study showed that accuracy of keeping the virtual RCM to predefined position is 3.0 mm. In a phantom test, the physicians found the needle insertion path faster with than without the manipulator (number of physicians = 3, P = 0.001). However, the alignment time with the virtual RCM was not shorter when imaging time for planning were considered. CONCLUSION: The study indicated that the robot holds promise as a tool for accurately and interactively selecting the optimal needle insertion path in liver ablation therapy guided by open configuration MRI. PMID- 18407543 TI - Gadolinium-enhanced echo-planar T2-weighted MRI of tumors in the extracranial head and neck: feasibility study and preliminary results using a distributed parameter tracer kinetic analysis. AB - PURPOSE: To examine the feasibility of first-pass dynamic contrast-enhanced (DCE) T2-weighted MRI of tumors in the extracranial head and neck by applying a distributed-parameter (DP) tracer kinetic model to quantify the perfusion parameters. MATERIALS AND METHODS: A total of 16 patients with primary malignant and benign tumors in the head and neck underwent DCE-MR studies. A spin-echo (SE) echo-planar-imaging (EPI) MR-sequence was applied for first-pass DCE-T2-weighted imaging. The data were postprocessed applying a DP tracer kinetic model that accounts for capillary-tissue exchange. Region-of-interest (ROI) analysis was performed in the tumor sites and the adjacent normal tissue. Blood flow (F), intravascular blood volume (v(1)), extravascular extracellular volume (v(2)), difference in bolus arrival time between arterial input and tissue (t(0)), intravascular mean transit time (t(1)), permeability (PS), and extraction ratio (E) maps were generated for each patient. RESULTS: All perfusion values in the tumor sites were significantly different (0.000 < or = P < or = 0.01) than those in the normal muscle tissue. The median perfusion values in the tumor tissue were: F = 150.5 mL/minute/100 g, v(1) = 11.0 mL/100 g, v(2) = 31.5 mL/100 g, t(0) = 4.5 seconds, t(1) = 8.0 seconds, PS = 96.0 mL/minute/100 g, and E = 32.5. CONCLUSION: EPI-T2-weighted DCE-MR in head and neck tumors as well as quantification of the perfusion values using DP model physiologic imaging was feasible and the promising initial results have encourages further validation studies in the future. PMID- 18407544 TI - Two-point Dixon fat-water separation: improving reliability and accuracy in phase correction algorithms. AB - PURPOSE: To propose an advanced phase-correction region-growing algorithm for two point fat-water separation suitable for parotid assessment, and to evaluate the general performance of phase-correction algorithms. MATERIALS AND METHODS: Two region-growing algorithms were evaluated in test objects and in head images: the original phase-correction algorithm (OPC) and the advanced phase-correction algorithm with voxel size manipulation (VSM) which includes: 1) starting the region-growing process from images of lower resolution and subsequently stepping toward the original matrix size, and 2) limiting the use of low-pass filters to fat-water interfaces with partial volume effects RESULTS: Fundamental problems relate to biological tissue spectrum being poorly approximated by two discrete peaks for fat and water. The VSM algorithm was shown to be less noise-sensitive, faster, and to produce a better approximation for the field inhomogeneity map. In head images (6 volunteers, 10 slices each) 43 errors were found with the OPC algorithm and only 6 errors with the VSM algorithm. Only the OPC algorithm produced errors surrounding the parotids (10 errors). CONCLUSION: The VSM algorithm provides a more accurate and less noise-sensitive fat-water separation. This highly significant performance improvement allows the application of phase correction algorithms to a wider range of clinical applications. PMID- 18407545 TI - Rapid magnetic resonance elastography of muscle using one-dimensional projection. AB - PURPOSE: To demonstrate the feasibility of 1D MR elastography (MRE) to rapidly assess skeletal muscle stiffness in vivo. MATERIALS AND METHODS: Shear waves were induced in the vastus medialis muscle (VM) using a pneumatic driver at 90 Hz and 2D MRE data were collected. Spatially selective excitations were used to produce 1D projections of MRE data oriented along the direction of propagating waves in the muscle. Data were collected with the thigh muscles relaxed and contracted at 20% maximum voluntary contraction (MVC) and the knee flexed at 30 degrees . RESULTS: The muscle stiffness measured at rest and in contraction with 1D MRE was 3.69 +/- 0.80 kPa and 9.52 +/- 2.74 kPa, respectively, and 4.36 +/- 0.98 kPa and 9.22 +/- 1.29 kPa, respectively, with the 2D MRE technique. CONCLUSION: Muscle stiffness measured using 1D MRE was in agreement with 2D MRE while reducing the scan time by a factor of 4. PMID- 18407546 TI - Publisher's note. Recently, concerns were raised regarding the possible nondisclosure of pertinent financial interests on the part of 2 scientists who were authors on 5 articles concerning lung cancer screening in the journals Cancer and Cancer Cytopathology from 2001 through 2007. PMID- 18407547 TI - Evolving into science advocates. PMID- 18407548 TI - The cholinergic gene locus in amphioxus: molecular characterization and developmental expression patterns. AB - The cholinergic gene locus (CGL), consisting of the vesicular acetylcholine transporter (VAChT)/choline acetyltransferase (ChAT) gene, encodes two specific cholinergic neuronal markers used extensively to study cholinergic transmission. In the present work, we isolated the amphioxus homologs of VAChT and ChAT and examined their expression during development. Analysis of the 5' untranslated region of VAChT and ChAT suggests that the splicing of the VAChT/ChAT mRNA has been evolutionarily conserved in amphioxus and mammals. By double whole-mount in situ hybridization, we demonstrate that VAChT and ChAT are coexpressed in the same cells. They are first expressed in four pairs of differentiating cells in the neural plate. Their later expression is primarily in the anterior nerve cord in several types of motoneurons, some of the interneurons and in the receptor cells of the larval ocellus. PMID- 18407549 TI - Autoregulatory loop and retinoic acid repression regulate pou2/pou5f1 gene expression in the zebrafish embryonic brain. AB - Zebrafish pou5f1, also known as pou2, encodes a POU-family transcription factor that is transiently expressed in the prospective midbrain and anterior hindbrain during gastrulation, governing brain development. In the present study, we found that the main regulatory elements reside in the proximal upstream DNA sequence from -2.2 to -0.12 kb (the -2.2/-0.1 region). The electrophoretic gel mobility shift assay (EMSA) revealed four functional octamer sequences that can associate with zebrafish Pou2/Pou5f1. The expression of mutated reporter constructs, as well as EMSA, suggested that these four octamer sequences operate in a cooperative manner to drive expression in the mid/hindbrain. We also identified a retinoic acid (RA)-responsive element in this proximal region, which was required to repress transcription in the posterior part of the embryo. These data provide a scheme wherein pou2/pou5f1 expression in zebrafish embryos is regulated by both an autoregulatory loop and repression by RA emanating from the posterior mesoderm. PMID- 18407550 TI - A novel mutation in AlphaA-crystallin (CRYAA) caused autosomal dominant congenital cataract in a large Chinese family. AB - Hereditary cataract is a phenotypically and genetically heterogeneous lens disease that is responsible for a significant proportion of the visual impairment and blindness that occurs in children. In a five-generation Chinese family with autosomal dominant inherited congenital cataract, clinical examination showed three cataract phenotypes: punctuate, nuclear, and total cataracts. Linkage analysis was performed and positive two-point LOD scores (with maximum of 4.43 and 4.27 at theta=0) were obtained for markers D21S1411 and D21S1890 on chromosome 21q22.3, flanking the CRYAA (alphaA-crystallin-encoding gene) locus. Sequencing of CRYAA revealed a novel heterozygous G>A transition (c.346G>A) in exon 3 that cosegregated with the disease phenotype and results in a conservative substitution of Arg to His at codon 116 (p.R116H). To understand the molecular basis of cataract formation, mutant and wild-type alphaA-crystallins were expressed in E. coli. RP-HPLC (reverse phase-high-performance liquid chromatography) suggested an increased hydrophobicity of the mutant recombinant protein, compared to that of wild-type alphaA-crystallins. Furthermore, loss of chaperone activity of the mutant was seen in DTT (DL-dithiothreitol)-induced insulin aggregation assay. FPLC (fast protein liquid chromatography) purification showed that the His-116 mutant protein had increased binding affinity to lysozyme. Gain of activated lysozyme binding, elevation of hydrophobicity and loss of chaperone activity of the mutant protein may be some of the molecular mechanisms underlying cataract in this large family. PMID- 18407551 TI - SORL1 is genetically associated with increased risk for late-onset Alzheimer disease in the Belgian population. AB - SORL1 has recently been identified as a major genetic contributor to increased risk for late-onset Alzheimer disease (AD). Here we aimed at replicating this finding in a large, well-characterized group of 550 Belgian late-onset AD patients and 637 healthy control individuals using a gene-wide genotyping approach across the SORL1 locus. We observed significant associations, both for individual SNPs (SNPs 6, 8, 9, 10 and 27; p-values ranging from 0.001 to 0.040) and 3-SNP haplotypes (SNPs 5-6-7 and SNPs 25-26-27; p-values ranging from 0.008 to 0.035). Moreover, the associations at SNP 8, 9 and 10 represented a direct replication of the initial association data. Two signals in distinct regions of the gene were shown to be mutually independent, supporting allelic heterogeneity at the SORL1 locus in the Belgian population. Our findings confirm that genetic variants in SORL1 may be important risk factors for late-onset AD. PMID- 18407552 TI - Homozygous mutation of the PHOX2B gene in congenital central hypoventilation syndrome (Ondine's Curse). AB - Homozygosity for a dominant allele is relatively rare and preferentially observed in communities with high inbreeding. According to the definition of true dominance, similar phenotypes should be observed in patients heterozygous and homozygous for a dominant mutation. However, the homozygous phenotype usually tends to be more severe than the heterozygous one. In these cases, the wild-type and mutant alleles are semi-dominant. Here we report a patient with a Congenital Central Hypoventilation Syndrome (CCHS) phenotype and homozygosity for a PHOX2B gene mutation leading to an alanine expansion shorter than the threshold hitherto observed in CCHS patients with a heterozygous mutation. This observation adds the concept of mutational threshold per se to the discussion about dominant and recessive alleles. PMID- 18407553 TI - The mutation p.Ser298Pro in the sulphamidase gene (SGSH) is associated with a slowly progressive clinical phenotype in mucopolysaccharidosis type IIIA (Sanfilippo A syndrome). AB - Mucopolysaccharidosis type IIIA (MPS IIIA, Sanfilippo A syndrome) is caused by mutations in the N-sulfoglucosamine sulfohydrolase (SGSH) gene and the resulting defective lysosomal degradation of the glycosaminoglycan heparan sulfate. The onset and progression of the disease are highly variable. Seventy-five mutations distributed over the SGSH gene have been described. We here report on the analysis of the natural course of the disease in 54 MPS IIIA patients through the use of a detailed questionnaire and four-point scoring system and an examination of the underlying mutations. By assessing the degree of developmental regression over time a group of seven patients with a slowly progressive course of the disease were identified. In these seven patients and in 3 other mildly affected patients the missense mutation c.892T>C (p.Ser298Pro) was found on one allele. These patients showed a lower frequency and later onset of the typical symptoms of the disease. The onset of regression in speech abilities and cognitive functions were delayed by 0.7 and 0.8 years, respectively, and the onset of regression of motor functions occurred 6.1 years later than in all other MPS IIIA patients. Severe regression in speech, cognitive and motor functions were delayed by 5, 5.9, and 11.2 years, respectively. These data suggest that in MPS IIIA patients carrying the mutation p.Ser298Pro a slowly progressive phenotype can be predicted and this may have an important impact on parental counselling and therapeutic interventions. PMID- 18407554 TI - Identification of genes involved in the ciliary trafficking of C. elegans PKD-2. AB - Ciliary membrane proteins are important extracellular sensors, and defects in their localization may have profound developmental and physiological consequences. To determine how sensory receptors localize to cilia, we performed a forward genetic screen and identified 11 mutants with defects in the ciliary localization (cil) of C. elegans PKD-2, a transient receptor potential polycystin (TRPP) channel. Class A cil mutants exhibit defects in PKD-2::GFP somatodendritic localization while Class B cil mutants abnormally accumulate PKD-2::GFP in cilia. Further characterization reveals that some genes mutated in cil mutants act in a tissue-specific manner while others are likely to play more general roles in such processes as intraflagellar transport (IFT). To this end, we identified a Class B mutation that disrupts the function of the cytoplasmic dynein light intermediate chain gene xbx-1. Identification of the remaining mutations will reveal novel molecular pathways required for ciliary receptor localization and provide further insight into mechanisms of ciliary signaling. PMID- 18407555 TI - Cholangiocyte primary cilia in liver health and disease. AB - The epithelial cells lining intrahepatic bile ducts (i.e., cholangiocytes), like many cell types in the body, have primary cilia extending from the apical plasma membrane into the bile ductal lumen. Cholangiocyte cilia express proteins such as polycystin-1, polycystin-2, fibrocystin, TRPV4, P2Y12, AC6, that account for ciliary mechano-, osmo-, and chemo-sensory functions; when these processes are disturbed by mutations in genes encoding ciliary-associated proteins, liver diseases (i.e., cholangiociliopathies) result. The cholangiociliopathies include but are not limited to cystic and fibrotic liver diseases associated with mutations in genes encoding polycystin-1, polycystin-2, and fibrocystin. In this review, we discuss the functions of cholangiocyte primary cilia, their role in the cholangiociliopathies, and potential therapeutic approaches. PMID- 18407556 TI - Control of gastrula cell motility by the Goosecoid/Mix.1/ Siamois network: basic patterns and paradoxical effects. AB - In the vegetal half of the Xenopus gastrula, cell populations differ with respect to migration on fibronectin substratum. We show that the paired-class homeodomain transcription factors Goosecoid (Gsc), Mix.1, and Siamois (Sia) are involved in the modulation of migration velocity and cell polarity. Mix.1 is expressed in the whole vegetal half and serves as a competence factor that is necessary, but not sufficient, for rapid cell migration and polarization. In the head mesoderm, Gsc and Sia are coexpressed with Mix.1, promoting rapid cell migration and polarization. Ectopic expression of Gsc and Sia in both vegetal and ventral regions often generates paradoxical effects; if a factor activates a certain motility trait in one region, it inhibits it in the other. Migration velocity and cell polarity are regulated independently. Fast and efficiently migrating multipolar cells and slow-moving polarized cells can be obtained by ectopic expression of these transcription factors in different combinations. PMID- 18407557 TI - Self-assembly of supramolecular luminescent Au(I)-Cu(I) complexes: "wrapping" an Au6Cu6 cluster in a [Au3(diphosphine)3]3+ "belt". PMID- 18407558 TI - High catalytic activity of dendritic C60 monoadducts in metal-free superoxide dismutation. PMID- 18407559 TI - Mutasynthesis of fluorosalinosporamide, a potent and reversible inhibitor of the proteasome. PMID- 18407560 TI - HIV/AIDS research. PMID- 18407561 TI - The essential role of balance tests in propensity-matched observational studies: comments on 'A critical appraisal of propensity-score matching in the medical literature between 1996 and 2003' by Peter Austin, Statistics in Medicine. PMID- 18407562 TI - Confidence intervals for ratios of AUCs in the case of serial sampling: a comparison of seven methods. AB - Pharmacokinetic studies are commonly performed using the two-stage approach. The first stage involves estimation of pharmacokinetic parameters such as the area under the concentration versus time curve (AUC) for each analysis subject separately, and the second stage uses the individual parameter estimates for statistical inference. This two-stage approach is not applicable in sparse sampling situations where only one sample is available per analysis subject similar to that in non-clinical in vivo studies. In a serial sampling design, only one sample is taken from each analysis subject. A simulation study was carried out to assess coverage, power and type I error of seven methods to construct two-sided 90% confidence intervals for ratios of two AUCs assessed in a serial sampling design, which can be used to assess bioequivalence in this parameter. PMID- 18407563 TI - Transition metal complexes of a cyclic pseudo hexapeptide: synthesis, complex formation and catalytic activities. AB - To contribute to a better understanding of metalloenzymes, we studied ion selectivity, complex formation tendencies and catalytic activities of linear and cyclic pseudopeptides. In this contribution, a linear and cyclic pseudo hexapeptide is described. The complex with transition metal ions and the sequence were designed using the programme COSMOS. Different routes of solid-phase synthesis were performed and compared using anchoring by C-terminus or a His side chain, using preformed pseudodipeptide building units or formation of N functionalized peptide bond during stepwise assembly. The different strategies were compared regarding cyclization tendency, yield and purity. Side-chain anchoring to solid support favours the cyclization but leads to the formation of difficult to separate dioxopiperazine. Both routes require preformed building units. Complex-formation tendencies and selectivity for certain bivalent transition metal ions were experimentally estimated and compared to ones predicted theoretically. CD measurements indicate conformational changes by complex formation with different metal ions. Catalytic activities on oxidation of catechol and hydrolysis of bis-phosphate esters by some metal complexes of linear and cyclic peptide show only low catalytic activities compared to other model peptides and related metalloenzymes. The preference of the cyclic peptide for complexation of Ni(2+) corresponds well to the predictions of COSMOS-NMR force field calculations. PMID- 18407564 TI - Nematicidal activity of anion transport blockers against Meloidogyne incognita, Caenorhabditis elegans and Heterorhabditis bacteriophora. AB - BACKGROUND: Because methyl bromide has been phased out as a soil sterilant, new nematicides are urgently needed. Four different chemical classes of organic acids acting as anion transport (AT) blockers were tested against a free-living nematode, Caenorhabditis elegans Maupas, a plant-parasitic nematode, Meloidogyne incognita (Kofoid and White) Chitwood, and an entomopathogenic nematode, Heterorhabditis bacteriophora Poinar, in toxicity bioassays. The materials tested were DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid), 9-AC (anthracene 9-carboxylic acid), NPPB [5-nitro-2-(3-phenylpropylamino)benzoic acid] and IAA-94 (indanyloxyacetic acid). RESULTS: All the compounds showed slowly developing nematicidal activity against second-stage juveniles of M. incognita and adults of C. elegans, but not against H. bacteriophora infective-stage juveniles. The LC(50) values of these compounds were < 50 mg L(-1) after 48 and 72 h incubation, while at 168 h incubation the LC(50) values were < 10 mg L(-1) for both sensitive species. Across both species and time, the LC(50) values generally differed no more than twofold among the four compounds tested in this study. In contrast, none of the compounds (200 mg L(-1)) caused more than control mortality to H. bacteriophora, even after 168 h of incubation. CONCLUSION: These compounds are potential leads for commercial nematicides. The insensitivity to H. bacteriophora is consistent with the natural exposure of this nematode to DST (3,5-dihydroxy-4 isopropylstilbene), a stilbene produced by its symbiotic bacterium. Based on the known activity of the compounds used in this study, it is suggested that anion transporters form the probable target sites for DIDS, 9-AC, NPPB and IAA-94 in nematodes. PMID- 18407565 TI - A 13C solid-state NMR analysis of steroid compounds. AB - (13)C CP/MAS solid-state NMR spectroscopy has been utilized to analyze six steroid compounds, namely testosterone (Tes), hydrocortisone (Cor), trans dehydroandrosterone (Adr), prednisolone (Prd), prednisone (Pre) and estradiol (Est). Among them, Tes displays a doublet pattern for all residues, whereas Prd, Pre and Est, exhibit exclusively singlets. For Cor and Adr, the (13)C spectra contain both doublet and singlet patterns. The (13)C doublet signal, with splittings of 0.2-1.5 ppm, are ascribed to local differences in the ring conformations associated with polymorphism. We have assigned all of the (13)C resonances to the different residues in these steroid compounds on the basis of solution NMR data. The C-7, C-8, C-10, C-15 and C-16 residues of Tes, Cor and Adr consistently give rise to singlets or doublets with splittings of less than 0.5 ppm, indicating similar local conformations. Accompanying hydration and dehydration processes, a reversible phase transformation between delta- and alpha crystal forms has been observed in Tes, corresponding to singlet and doublet (13)C patterns, respectively. To further characterize the ring conformations in the alpha-form, we have successfully extracted chemical shift tensor elements for the (13)C doublets. It is demonstrated that (13)C solid-state NMR spectroscopy provides a reliable and sensitive means of characterizing polymorphism in steroids. PMID- 18407566 TI - Further conventions for NMR shielding and chemical shifts (IUPAC Recommendations 2008). AB - IUPAC has published a number of recommendations regarding the reporting of nuclear magnetic resonance (NMR) data, especially chemical shifts. The most recent publication [Pure Appl. Chem. 73, 1795 (2001)] recommended that tetramethylsilane (TMS) serve as a universal reference for reporting the shifts of all nuclides, but it deferred recommendations for several aspects of this subject. This document first examines the extent to which the (1)H shielding in TMS itself is subject to change by variation in temperature, concentration, and solvent. On the basis of recently published results, it has been established that the shielding of TMS in solution [along with that of sodium-3 (trimethylsilyl)propanesulfonate, DSS, often used as a reference for aqueous solutions] varies only slightly with temperature but is subject to solvent perturbations of a few tenths of a part per million (ppm). Recommendations are given for reporting chemical shifts under most routine experimental conditions and for quantifying effects of temperature and solvent variation, including the use of magnetic susceptibility corrections and of magic-angle spinning (MAS). This document provides the first IUPAC recommendations for referencing and reporting chemical shifts in solids, based on high-resolution MAS studies. Procedures are given for relating (13)C NMR chemical shifts in solids to the scales used for high-resolution studies in the liquid phase. The notation and terminology used for describing chemical shift and shielding tensors in solids are reviewed in some detail, and recommendations are given for best practice. PMID- 18407567 TI - Multiple diseases in carrier probability estimation: accounting for surviving all cancers other than breast and ovary in BRCAPRO. AB - Mendelian models can predict who carries an inherited deleterious mutation of known disease genes based on family history. For example, the BRCAPRO model is commonly used to identify families who carry mutations of BRCA1 and BRCA2, based on familial breast and ovarian cancers. These models incorporate the age of diagnosis of diseases in relatives and current age or age of death. We develop a rigorous foundation for handling multiple diseases with censoring. We prove that any disease unrelated to mutations can be excluded from the model, unless it is sufficiently common and dependent on a mutation-related disease time. Furthermore, if a family member has a disease with higher probability density among mutation carriers, but the model does not account for it, then the carrier probability is deflated. However, even if a family only has diseases the model accounts for, if the model excludes a mutation-related disease, then the carrier probability will be inflated. In light of these results, we extend BRCAPRO to account for surviving all non-breast/ovary cancers as a single outcome. The extension also enables BRCAPRO to extract more useful information from male relatives. Using 1500 families from the Cancer Genetics Network, accounting for surviving other cancers improves BRCAPRO's concordance index from 0.758 to 0.762 (p=0.046), improves its positive predictive value from 35 to 39 per cent (p<10( 6)) without impacting its negative predictive value, and improves its overall calibration, although calibration slightly worsens for those with carrier probability<10 per cent. PMID- 18407569 TI - Synthesis and complete assignment of the 1H and 13C NMR signals of some oxopyrancoumarin and oxofuropyrancoumarin derivatives. AB - The synthesis of four pyranocoumarins starting from phloroglucinol and the complete (1)H and (13)C NMR assignment of seven pyranocoumarins has been performed using 1D and 2D NMR techniques including COSY, HMQC and HMBC experiments. PMID- 18407568 TI - The generalized F distribution: an umbrella for parametric survival analysis. AB - In a recent tutorial my colleagues and I advocated the generalized gamma (GG) distribution as a platform for parametric survival analysis. The GG family includes all four of the common types of hazard functions, making it particularly useful for estimating individual hazard functions as well as both relative hazards and relative times. In addition, the GG includes most of the commonly used parametric survival distributions. Survival analysis based on the GG distribution is practical since regression models are available in commonly used statistical packages. It is well known that the GG is contained in an even larger family, the generalized F (GF) distribution, which also includes the log logistic. The GF thus provides additional flexibility for parametric modeling. In this paper we discuss the GF family from this perspective. We provide a characterization of the hazard functions of the GF, showing that, except for the GG, the available hazard functions are limited to decreasing and arc-shaped hazards and, in particular, that the hazard function can be decreasing but not monotone. We also discuss fitting the GF with an alternative parameterization using standard statistical software and refine a description of the hazard functions for death after a diagnosis of clinical AIDS in four different eras of HIV therapy. PMID- 18407570 TI - Spatial clustering of the failure to geocode and its implications for the detection of disease clustering. AB - Geocoding a study population as completely as possible is an important data assimilation component of many spatial epidemiologic studies. Unfortunately, complete geocoding is rare in practice. The failure of a substantial proportion of study subjects' addresses to geocode has consequences for spatial analyses, some of which are not yet fully understood. This article explicitly demonstrates that the failure to geocode can be spatially clustered, and it investigates the implications of this for the detection of disease clustering. A data set of more than 9000 ground-truthed addresses from Carroll County, Iowa, which was geocoded via a standard address matching and street interpolation algorithm, is used for this purpose. Through simulation of disease processes at these addresses, the authors show that spatial clustering of geocoding failure has no effect on the marginal power to detect spatial disease clustering if the likelihood of disease is independent of the failure to geocode, but that power is substantially reduced if disease likelihood and geocoding failure are positively associated. PMID- 18407571 TI - A distribution-free test of constant mean in linear mixed effects models. AB - We propose a distribution-free procedure, an analogy of the DIP test in non parametric regression, to test whether the means of responses are constant over time in repeated measures data. Unlike the existing tests, the proposed procedure requires very minimal assumptions to the distributions of both random effects and errors. We study the asymptotic reference distribution of the test statistic analytically and propose a permutation procedure to approximate the finite-sample reference distribution. The size and power of the proposed test are illustrated and compared with competitors through several simulation studies. We find that it performs well for data of small sizes, regardless of model specification. Finally, we apply our test to a data example to compare the effect of fatigue in two different methods used for cardiopulmonary resuscitation. PMID- 18407572 TI - A simulation-based comparison of techniques to correct for measurement error in matched case-control studies. AB - The presence of measurement errors affecting the covariates in regression models is a relevant topic in many scientific areas, as, for example, in epidemiology. An example is given by an epidemiological population-based matched case-control study on the aetiology of childhood malignancies, which is currently under completion in Italy. This study was aimed at evaluating the effects of childhood exposure to extremely low electromagnetic fields on the risk of disease occurrence by taking into account the possibility of erroneous measures of the exposure. Within this framework, we focus on the application of likelihood methods to correct for measurement error. This approach, which has received less attention in literature with respect to alternatives, is compared with commonly used methods such as regression calibration and SIMEX. The comparison is performed by simulation, under a broad range of measurement error structures. PMID- 18407573 TI - An index approach for the Cox model with left censored covariates. AB - Medical studies frequently collect biological markers in which many subjects have values below the detectable limits of the assay, resulting in heavily censored data. We develop a modification of the Rigobon and Stoker index method for application to a Cox regression model with censored covariates. The index approach is compared with a complete case method and various fill-in methods. Our simulation results demonstrated that the index approach is an improvement over the other methods. We illustrated the usefulness of this approach with an example for the GenIMS study examining the relationship between two inflammatory markers and survival. PMID- 18407574 TI - Interval estimation of risk difference for data sampled from clusters. AB - Risk difference (RD) is an important measure in epidemiological studies where the probability of developing a disease for individuals in an exposed group, for example, is compared with that in a control group. There are varying cluster sizes in each group and the binary responses within each cluster cannot be assumed independent. Under the cluster sampling scenario, Lui (Statistical Estimation of Epidemiological Risk. Wiley: CA, 2004; 7-27) discusses four methods for the construction of a confidence interval for the RD. In this paper we introduce two very simple methods. One method is based on an estimator of the variance of a ratio estimator (Sampling Techniques (3rd edn). Wiley: New York, 1977; 30-67) and the other method is based on a sandwich estimator of the variance of the regression estimator using the generalized estimating equations approach of Zeger and Liang (Biometrics 1986; 42:121-130). These two methods are then compared, by simulation, in terms of maintaining nominal coverage probability and average coverage length, with the four methods discussed by Lui (Statistical Estimation of Epidemiological Risk. Wiley: CA, 2004; 7-27). Simulations show at least as good properties of these two methods as those of the others. The method based on an estimate of the variance of a ratio estimator performs best overall. It involves a very simple variance expression and can be implemented with a very few computer codes. Therefore, it can be considered as an easily implementable alternative. PMID- 18407575 TI - A note on the conservativeness of the confidence interval approach for the selection of non-inferiority margin in the two-arm active-control trial. AB - Compared with placebo-control clinical trials, the interpretation of efficacy results from active-control trials requires more caution. This is because efficacy results from such trials cannot be reliably interpreted without a thorough understanding of the efficacy evidence that formed the basis for the approval of the active control, especially when such drug efficacy is to be established on the basis of clinical evidence from the traditional two-arm active control clinical equivalence studies as opposed to the multi-arm active control. This is because in addition to over-reliance on the quantification of a clinically irrelevant acceptable margin of inferiority from historical data, such interpretation also depends on cross-trial inference for demonstration of experimental drug effect. We provide a brief overview of some design issues with the traditional two-arm active-control clinical trial and discuss regulators' concern regarding Type I error rate control (with the two most popular methods for the quantification of the non-inferiority margin) in cross-trial demonstration of experimental drug effect. Simulation results are presented to show that the point estimate method provides adequate control of the Type I error rate with > or =75 per cent retention of known active-control effect and that the confidence interval approach is uniformly ultra-conservative. We also report (via a numerical example from real clinical trial data) a couple of potentially less stringent alternative approaches for establishing the non-inferiority of a test drug over a control, which have been used in the past to provide additional efficacy evidence in NDA submission. PMID- 18407577 TI - A note on 'The method of handling non-compliance in clinical trials suggested by Sato/Matsuyama'. PMID- 18407576 TI - Assessing the utility of public health surveillance using specificity, sensitivity, and lives saved. AB - In modern surveillance of public health, data may be reported in a timely fashion and include spatial data on cases in addition to the time of their occurrence. This has lead to many recent developments in statistical methods to detect events of public health importance. However, there has been relatively little work about how to compare such methods. One powerful rationale for performing surveillance is earlier detection of events of public health significance; previous evaluation tools have focused on metrics that include the timeliness of detection in addition to sensitivity and specificity. However, such metrics have not accounted for the number of persons affected by the events. We re-examine the rationale for this surveillance and conclude that earlier detection is preferred because it can prevent additional morbidity and mortality. On the basis this observation, we propose evaluating the number of cases prevented by each detection method, and include this information in assessing the value of different detection methods. Using this approach incorporates more information about the events and the detection and provides a sound basis for making decisions about which detection methods to employ. PMID- 18407578 TI - Confidence intervals for P(Y1>Y2) with normal outcomes in linear models. AB - Recently, there is an emerging interest in the inference of P(Y1>Y2) where Y1 and Y2 stand for two independent continuous random variables. So far, most of the research in this field focuses on simply comparing two outcomes without adjusting for covariates. This paper mainly presents a large sample approach based on a noncentral t distribution for the confidence interval estimation of P(Y1>Y2) with normal outcomes in linear models. Furthermore, the performance of the proposed large sample approach is compared with that of a generalized variable approach and a bootstrap approach. Simulation studies demonstrate that for small-to-medium sample sizes, both the large sample approach and the generalized variable approach provide confidence intervals with satisfying coverage probabilities whereas the bootstrap approach can be slightly liberal for certain scenarios. The proposed approaches are applied to three real-life data sets. PMID- 18407579 TI - Mechanistic studies of amide bond scission during acidolytic deprotection of Pip containing peptide. AB - Unusual TFA catalyzed cleavage reaction is reported for peptide containing pipecolic acid residues. Although the use of TFA under standard cleavage conditions is sufficiently mild to prevent degradation of the desired products, the amide bond between consecutive pipecolic acid residues is unexpectedly hydrolyzed by standard TFA treatment. The hydrolysis is proposed to proceed via an oxazolinium ion intermediate. This mechanism is supported by H/D exchange as observed by ESI-MS and NMR experiments. PMID- 18407580 TI - Analyzing sequentially randomized trials based on causal effect models for realistic individualized treatment rules. AB - In this paper, we argue that causal effect models for realistic individualized treatment rules represent an attractive tool for analyzing sequentially randomized trials. Unlike a number of methods proposed previously, this approach does not rely on the assumption that intermediate outcomes are discrete or that models for the distributions of these intermediate outcomes given the observed past are correctly specified. In addition, it generalizes the methodology for performing pairwise comparisons between individualized treatment rules by allowing the user to posit a marginal structural model for all candidate treatment rules simultaneously. This is particularly useful if the number of such rules is large, in which case an approach based on individual pairwise comparisons would be likely to suffer from too much sampling variability to provide an informative answer. In addition, such causal effect models represent an interesting alternative to methods previously proposed for selecting an optimal individualized treatment rule in that they immediately give the user a sense of how the optimal outcome is estimated to change in the neighborhood of the identified optimum. We discuss an inverse-probability-of-treatment-weighted (IPTW) estimator for these causal effect models, which is straightforward to implement using standard statistical software, and develop an approach for constructing valid asymptotic confidence intervals based on the influence curve of this estimator. The methodology is illustrated in two simulation studies that are intended to mimic an HIV/AIDS trial. PMID- 18407581 TI - Direct injection LC/ESI-MS horse urine analysis for the quantification and identification of threshold substances for doping control. I. Determination of hydrocortisone. AB - Two simple and rapid LC/MS methods with direct injection analysis were developed and validated for the quantification and identification of hydrocortisone in equine urine using the same sample preparation but different mass spectrometric systems: ion trap mass spectrometry (IT-MS) and time-of-flight mass spectrometry (TOF-MS). The main advantage of the proposed methodology is the minimal sample preparation procedure, as particle-free diluted urine samples were directly injected into both LC/MS systems. Desonide was used as internal standard (IS). The linear range was 0.25-2.5 microg ml(-1) for both methods. Matrix effects were evaluated by preparing and analyzing calibration curves in water solutions and different horse urine samples. A great variation of the signal both for hydrocortisone and the internal standard was observed in different matrices. To overcome matrix effects, the unavailability of blank matrix and the excessive cost of the isotopically labeled internal standard, standard additions calibration method was applied. This work is an exploration of the performance of the standard additions approach in a method where neither nonisotopic internal standards nor extensive sample preparation is utilized and no blank matrix is available. The relative standard deviations of intra and interday analysis of hydrocortisone in horse urine were lower than 10.2 and 5.4%, respectively, for the LC/IT-MS method and lower than 8.4 and 4.4%, respectively, for the LC/TOF-MS method. Accuracy (bias percentage) was less than 9.7% for both methods. PMID- 18407582 TI - Parametric robust inferences for correlated ordinal data. AB - The aim of this article is to provide asymptotically valid likelihood inferences about regression parameters for correlated ordinal response variables. The legitimacy of this novel approach requires no knowledge of the underlying joint distributions so long as their second moments exist. The efficacy of the proposed parametric approach is demonstrated via simulations and the analyses of two real data sets. PMID- 18407583 TI - Identifying significant covariates for anti-HIV treatment response: mechanism based differential equation models and empirical semiparametric regression models. AB - In this paper, the mechanism-based ordinary differential equation (ODE) model and the flexible semiparametric regression model are employed to identify the significant covariates for antiretroviral response in AIDS clinical trials. We consider the treatment effect as a function of three factors (or covariates) including pharmacokinetics, drug adherence and susceptibility. Both clinical and simulated data examples are given to illustrate these two different kinds of modeling approaches. We found that the ODE model is more powerful to model the mechanism-based nonlinear relationship between treatment effects and virological response biomarkers. The ODE model is also better in identifying the significant factors for virological response, although it is slightly liberal and there is a trend to include more factors (or covariates) in the model. The semiparametric mixed-effects regression model is very flexible to fit the virological response data, but it is too liberal to identify correct factors for the virological response; sometimes it may miss the correct factors. The ODE model is also biologically justifiable and good for predictions and simulations for various biological scenarios. The limitations of the ODE models include the high cost of computation and the requirement of biological assumptions that sometimes may not be easy to validate. The methodologies reviewed in this paper are also generally applicable to studies of other viruses such as hepatitis B virus or hepatitis C virus. PMID- 18407584 TI - Modeling heaping in self-reported cigarette counts. AB - In studies of smoking behavior, some subjects report exact cigarette counts, whereas others report rounded-off counts, particularly multiples of 20, 10 or 5. This form of data reporting error, known as heaping, can bias the estimation of parameters of interest such as mean cigarette consumption. We present a model to describe heaped count data from a randomized trial of bupropion treatment for smoking cessation. The model posits that the reported cigarette count is a deterministic function of an underlying precise cigarette count variable and a heaping behavior variable, both of which are at best partially observed. To account for an excess of zeros, as would likely occur in a smoking cessation study where some subjects successfully quit, we model the underlying count variable with zero-inflated count distributions. We study the sensitivity of the inference on smoking cessation by fitting various models that either do or do not account for heaping and zero inflation, comparing the models by means of Bayes factors. Our results suggest that sufficiently rich models for both the underlying distribution and the heaping behavior are indispensable to obtaining a good fit with heaped smoking data. The analyses moreover reveal that bupropion has a significant effect on the fraction abstinent, but not on mean cigarette consumption among the non-abstinent. PMID- 18407585 TI - Value of colonoscopy for prediction of prognosis in patients with ulcerative colitis. AB - Ulcerative colitis (UC) is a chronic inflammatory bowel disorder characterized by exacerbations and remissions. Some UC patients remain refractory to conventional medical treatment while, in others, the effectiveness of drugs is limited by side effects. Recently, cyclosporine and leukocyte removal therapy have been used for refractory UC patients. To predict the efficacy of these therapies is important for appropriate selection of treatment options and for preparation for colectomy. Endoscopy is the cornerstone for diagnosis and evaluation of UC. Endoscopic parameters in patients with severe or refractory UC may predict a clinical response to therapies, such as cyclosporine or leukocyte removal therapy. As for the patients with quiescent UC, relapse of UC is difficult to predict by routine colonoscopy. Even when routine colonoscopy suggests remission and a normal mucosal appearance, microscopic abnormalities may persist and relapse may occur later. To more accurately identify disease activity and to predict exacerbations in UC patients with clinically inactive disease is important for deciding whether medical treatment should be maintained. Magnifying colonoscopy is useful for the evaluation of disease activity and for predicting relapse in patients with UC. PMID- 18407586 TI - MUC1 and colorectal cancer pathophysiology considerations. AB - Several lines of evidence point towards a biological role of mucin and particularly MUC1 in colorectal cancer. A positive correlation was described between mucin secretion, proliferation, invasiveness, metastasis and bad prognosis. But, the role of MUC1 in cancer progression is still controversial and somewhat confusing. While Mukherjee and colleagues developed MUC1-specific immune therapy in a CRC model, Lillehoj and co-investigators showed recently that MUC1 inhibits cell proliferation by a beta-catenin-dependent mechanism. In carcinoma cells the polarization of MUC1 is lost and the protein is over expressed at high levels over the entire cell surface. A competitive interaction between MUC1 and E cadherin, through beta-catenin binding, disrupts E-cadherin-mediated cell-cell interactions at sites of MUC1 expression. In addition, the complex of MUC1-beta catenin enters the nucleus and activates T-cell factor/leukocyte enhancing factor 1 transcription factors and activates gene expression. This mechanism may be similar to that just described for DCC and UNC5H, which induced apoptosis when not engaged with their ligand netrin, but mediate signals for proliferation, differentiation or migration when ligand bound. PMID- 18407588 TI - Effect of NHE1 antisense gene transfection on the biological behavior of SGC-7901 human gastric carcinoma cells. AB - AIM: To study the effect of type 1 Na(+)/H(+) exchanger (NHE1) antisense human gene transfection on the biological behavior of gastric carcinoma cell line SGC 7901. METHODS: Antisense NHE1 eukaryotic expression on vector pcDNA3.1 was constructed by recombinant DNA technique and transfected into gastric carcinoma cell line SGC-7901 with DOTAP liposome transfection method. Morphological changes of cells were observed with optic and electron microscopes. Changes in cell proliferative capacity, apoptosis, intracellular pH (pH(i)), cell cycle, clone formation in two-layer soft agar, and tumorigenicity in nude mice were examined. RESULTS: Antisense eukaryotic expressing vectors were successfully constructed and transfected into SGC-7901. The transfectant obtained named 7901-antisense (7901-AS) stablely produced antisense NHE1. There was a significant difference between the pH(i) of 7901-AS cells (6.77 +/- 0.05) and that of 7901-zeo cells and SGC-7901 cells (7.24 +/- 0.03 and 7.26 +/- 0.03, P < 0.01). Compared with SGC 7901 and 7901-zeo cells, 7901-AS cells mostly showed cell proliferation inhibition, G1/G0 phase arrest, increased cell apoptotic rate, recovery of contact inhibition, and density contact. The tumorigenicity in nude mice and cloning efficiency in the two-layer soft agar were clearly inhibited. CONCLUSION: NHE1 antisense gene significantly restrains the malignant behavior of human gastric carcinoma cells, suppresses cell growth and induces cell apoptosis, and partially reverses the malignant phenotypes of SGC-7901. These results suggest a potential role for human tumor gene therapy. PMID- 18407587 TI - Necrotizing enterocolitis: a multifactorial disease with no cure. AB - Necrotizing enterocolitis is an inflammatory bowel disease of neonates with significant morbidity and mortality in preterm infants. Due to the multifactorial nature of the disease and limitations in disease models, early diagnosis remains challenging and the pathogenesis elusive. Although preterm birth, hypoxic ischemic events, formula feeding, and abnormal bacteria colonization are established risk factors, the role of genetics and vasoactive/inflammatory mediators is unclear. Consequently, treatments do not target the specific underlying disease processes and are symptomatic and surgically invasive. Breast feeding is the most effective preventative measure. Recent advances in the prevention of necrotizing enterocolitis have focused on bioactive nutrients and trophic factors in human milk. Development of new disease models including the aspect of prematurity that consistently predisposes neonates to the disease with multiple risk factors will improve our understanding of the pathogenesis and lead to discovery of innovative therapeutics. PMID- 18407589 TI - Troglitazone, a peroxisome proliferator-activated receptor gamma ligand, induces growth inhibition and apoptosis of HepG2 human liver cancer cells. AB - AIM: To examine the effect of troglitazone, a peroxisome proliferator-activated receptor gamma (PPAR gamma) ligand, on the proliferation and apoptosis of human liver cancer cells. METHODS: Liver cancer cell line HepG2 was cultured and treated with troglitazone. Cell proliferation was detected by 3-(4-,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; apoptosis was detected by flow cytometry and terminal deoxynucleotidyl transferase-mediated nick end labeling of DNA fragmentation sites (TUNEL) assay; and apoptosis-related protein was detected by immunocytochemistry and Western blotting. RESULTS: Troglitazone inhibited growth and induced apoptosis of HepG2 cells in a dose dependent manner, and induced activation of caspase-3 expression. Troglitazone not only drove apoptosis-inhibiting factor survivin to translocate incompletely from the nucleus to the cytoplasm, but also inhibited expression of survivin, while it did not affect expression of apoptosis-promoting factor Bax. CONCLUSION: PPAR gamma ligands inhibit growth and induce apoptosis of liver cancer cells, and may have applications for the prevention and treatment of liver cancer. PMID- 18407590 TI - Detection of apoptosis induced by new type gosling viral enteritis virus in vitro through fluorescein annexin V-FITC/PI double labeling. AB - AIM: To achieve a better understanding of the pathogenesis of new type gosling viral enteritis virus (NGVEV) and the relationship between NGVEV and host cells. METHODS: The apoptosis of duck embryo fibroblasts (DEF) induced by NGVEV was investigated by fluorescence-activated cell sorter (FACS) and fluorescence microscope after the cells were stained with Annexin V-FITC and propidium iodide (PI). RESULTS: By staining cells with a combination of fluorescein annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze non-apoptotic cells (Annexin V-FITC negative/PI negative), early apoptotic cells (Annexin V FITC positive/PI negative), late apoptotic/necrotic cells (Annexin V-FITC positive/PI positive) and dead cells (Annexin V-FITC negative/PI positive) through flow cytometry and fluorescence microscope. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 120 h after infection, while the percentage of non-apoptotic cells decreased. CONCLUSION: NGVEV can induce the infected DEF cells to undergo apoptosis and the apoptosis occurs prior to necrosis. PMID- 18407591 TI - Pharmacokinetics and tissue distribution of intraperitoneal 5-fluorouracil with a novel carrier solution in rats. AB - AIM: To compare the pharmacokinetics and tissue distribution of 5-fluorouracil administered intraperitoneally with two isotonic carrier solutions: HAES-steri (neotype 6% hydroxyethyl starch), a novel carrier solution with middle molecular weight and physiologic saline (0.9% sodium chloride solution), a traditional carrier solution for intraperitoneal chemotherapy, in rats. METHODS: A total of 60 Sprague Dawley rats were randomized into groups according to the carrier solution administered. Each group was further randomized according to the intraperitoneal dwell period (1, 3, 6, 12, 18 and 24 h). At the end of the procedure the rats were killed, the peritoneal fluid was withdrawn completely and quantitated. Drug concentrations in peritoneal fluid, plasma, and tissues were determined by high-performance liquid chromatography. RESULTS: The mean volumes remaining in the peritoneal cavity were significantly higher with HAES-steri than those with physiologic saline at 1, 6, 12, 18, and 24 h (P = 0.047, 0.009, 0.005, 0.005 and 0.005 respectively, the percentages of remaining peritoneal fluid volume were 89.9 +/- 5.6 vs 83.4 +/- 4.9, 79.9 +/- 2.8 vs 56.2 +/- 15.7, 46.8 +/- 5.5 vs 24.7 +/- 9.7, 23.0 +/- 2.8 vs 0.0 +/- 0.0 and 4.2 +/- 1.7 vs 0.0 +/- 0.0 respectively). Mean concentrations in peritoneal fluid were significantly higher with HAES-steri than those with physiologic saline at 3, 12 and 18 h (P = 0.009, 0.009 and 0.005 respectively, the concentrations were 139.2768 +/- 28.2317 mg/L vs mg/L, 11.5427 +/- 3.0976 mg/L vs 0.0000 +/- 0.0000 mg/L and 4.7724 +/- 1.0936 mg/L vs 0.0000 +/- 0.0000 mg/L respectively). Mean plasma 5-fluorouracil concentrations in portal vein were significantly higher with HAES-steri at 3, 12, 18 and 24 h (P = 0.009, 0.034, 0.005 and 0.019 respectively, the concentrations were 3.3572 +/- 0.8128 mg/L vs 0.8794 +/- 0.2394 mg/L, 0.6203 +/- 0.9935 mg/L vs 0.0112 +/- 0.0250 mg/L, 0.3725 +/- 0.3871 mg/L vs 0.0000 +/- 0.0000 mg/L, and 0.2469 +/- 0.1457 mg/L vs 0.0000 +/- 0.0000 mg/L respectively), but significantly lower at 1 h (P = 0.009, the concentrations were 4.1957 +/- 0.6952 mg/L vs 7.7406 +/- 1.2377 mg/L). There were no significant differences in the plasma 5 fluorouracil in inferior caval vein at each time-point. 5-fluorouracil concentrations were significantly greater with HAES-steri at 18 h in gastric tissue (P = 0.016, the concentrations were 0.9486 +/- 0.8173 mg/L vs 030392 +/- 0.0316 mg/L), at 18 h in colon (P = 0.009, the concentrations were 0.1730 +/- 0.0446 mg/L vs 0.0626 +/- 0.0425 mg/L), at 3, 6, 12 and 24 h in liver (P = 0.009, 0.013, 0.034 and 0.013 respectively, the concentrations were 0.6472685 +/- 0.5256 mg/L vs 0.1554 +/- 0.1043mg/L, 0.8606826 +/- 0.7155 mg/L vs 0.0014 +/- 0.0029 mg/L, 0.0445 +/- 0.0330 mg/L vs 0.0797 +/- 0.1005 mg/L and 0.0863 +/- 0.0399 mg/L vs 0.0034 +/- 0.0075 mg/L respectively) and at 18 h in lung (P = 0.009, the concentrations were 0.0886 +/- 0.0668 mg/L vs 0.0094 +/- 0.0210 mg/L). There were no differences in 5-fluorouracil concentrations in renal tissue at each time point. CONCLUSION: The use of intraperitoneal 5-fluoro-uracil with HAES-Steri carrier solution provides a pharmacokinetic advantage for a local-regional killing of residual tumor cells and improve the accumulated penetrability of 5 fluorouracil with decreased systemic toxicity. Further clinical feasibility studies on the use of HAES-steri as carrier solution for intraperitoneal chemotherapy with 5-fluorouracil are warranted. PMID- 18407592 TI - Prospective cohort comparison of flavonoid treatment in patients with resected colorectal cancer to prevent recurrence. AB - AIM: To investigate biological prevention with flavonoids the recurrence risk of neoplasia was studied in patients with resected colorectal cancer and after adenoma polypectomy. METHODS: Eighty-seven patients, 36 patients with resected colon cancer and 51 patients after polypectomy, were divided into 2 groups: one group was treated with a flavonoid mixture (daily standard dose 20 mg apigenin and 20 mg epigallocathechin-gallat, n = 31) and compared with a matched control group (n = 56). Both groups were observed for 3-4 years by surveillance colonoscopy and by questionnaire. RESULTS: Of 87 patients enrolled in this study, 36 had resected colon cancer and 29 of these patients had surveillance colonoscopy. Among the flavonoid-treated patients with resected colon cancer (n = 14), there was no cancer recurrence and one adenoma developed. In contrast the cancer recurrence rate of the 15 matched untreated controls was 20% (3 of 15) and adenomas evolved in 4 of those patients (27%). The combined recurrence rate for neoplasia was 7% (1 of 14) in the treated patients and 47% (7 of 15) in the controls (P = 0.027). CONCLUSION: Sustained long-term treatment with a flavonoid mixture could reduce the recurrence rate of colon neoplasia in patients with resected colon cancer. PMID- 18407593 TI - A red wine polyphenolic extract reduces the activation phenotype of cultured human liver myofibroblasts. AB - AIM: To test the effect of a standardized red wine polyphenolic extract (RWPE) on the phenotype of human liver myofibroblasts in culture. METHODS: Human myofibroblasts grown from liver explants were used in this study. Cell proliferation was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Signaling events were analyzed by western blot with phospho-specific antibodies. Matrix-metalloproteinase activity was measured with gel zymography. RESULTS: We found that cell proliferation was dose-dependently decreased by up to 90% by RWPE while cell viability was not affected. Exposure to RWPE also greatly decreased the phosphorylation of ERK1/ERK2 and Akt in response to stimulation by the mitogenic factor platelet derived growth factor BB (PDGF-BB). Finally, RWPE affected extracellular matrix remodeling by decreasing the secretion by myofibroblasts of matrix metalloproteinase-2 and of tissue inhibitor of matrix-metalloproteinases-1. CONCLUSION: Altogether, RWPE decreases the activation state of liver myofibroblasts. The identification of the active compounds in RWPE could offer new therapeutic strategies against liver fibrosis. PMID- 18407594 TI - Effects of estradiol and progesterone on the proinflammatory cytokine production by mononuclear cells from patients with chronic hepatitis C. AB - AIM: To investigate the effects of estradiol (E2) and progesterone on the unstimulated and oxidative stress-stimulated production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-8, and macrophage chemotactic protein (MCP)-1 by peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis C and healthy controls. METHODS: The PBMCs were separated from age matched 72 males and 71 females with and without chronic hepatitis C, who were divided into two groups based on a mean menopausal age of 50 years. Oxidative stress was induced by hydrogen peroxide in the cells incubated in serum-free media. Cytokines in the culture supernatant were measured by an enzyme-linked immunosorbent assay. RESULTS: The highest levels of the spontaneous production of TNF-alpha, IL-1beta, IL-8, and MCP-1 by the unstimulated PBMCs were in the older male patients with chronic hepatitis C and the lowest levels were in the pre menopausal female healthy controls. E2 inhibited the cytokine production by the unstimulated PBMCs from the older male and post-menopausal female patients, which was further stimulated by progesterone. The exposure to hydrogen peroxide in the PBMCs from the younger male and pre-menopausal female healthy subjects induced the production of cytokines. The change rates of the hydrogen peroxide-stimulated cytokine production were suppressed by E2 and enhanced by progesterone. CONCLUSION: These findings suggest that E2 may play a favorable role in the course of persistent liver injury by preventing the accumulation of monocytes macrophages and by inhibiting proinflammatory cytokine production, whereas progesterone may counteract the favorable E2 effects. PMID- 18407595 TI - Comparison of CT and MRI for presurgical characterization of paraaortic lymph nodes in patients with pancreatico-biliary carcinoma. AB - AIM: To determine the accuracy of computed tomography (CT) and magnetic resonance (MR) for presurgical characterization of paraaortic lymph nodes in patients with pancreatico-biliary carcinoma. METHODS: Two radiologists independently evaluated CT and MR imaging of 31 patients who had undergone lymphadenectomy (9 metastatic and 22 non-metastatic paraaortic nodes). Receiver operating characteristic (ROC) curve analysis was performed using a five point scale to compare CT with MRI. To re-define the morphologic features of metastatic nodes, we evaluated CT scans from 70 patients with 23 metastatic paraaortic nodes and 47 non-metastatic ones. The short axis diameter, ratio of the short to long axis, shape, and presence of necrosis were compared between metastatic and non-metastatic nodes by independent samples t-test and Fisher's exact test. P < 0.05 was considered statistically significant. RESULTS: The mean area under the ROC curve for CT (0.732 and 0.646, respectively) was slightly higher than that for MRI (0.725 and 0.598, respectively) without statistical significance (P = 0.940 and 0.716, respectively). The short axis diameter of the metastatic lymph nodes (mean = 9.2 mm) was significantly larger than that of non-metastatic ones (mean = 5.17 mm, P < 0.05). Metastatic nodes had more irregular margins (44.4%) and central necrosis (22.2%) than non-metastatic ones (9% and 0%, respectively), with statistical significance (P < 0.05). CONCLUSION: The accuracy of CT scan for the characterization of paraaortic nodes is not different from that of MRI. A short axis-diameter (> 5.3 mm), irregular margin, and presence of central necrosis are the suggestive morphologic features of metastatic paraaortic nodes. PMID- 18407596 TI - Thrombospondin-1 expression correlates with angiogenesis in experimental cirrhosis. AB - AIM: To investigate the significance of Thrombospondin-1 (TSP-1) expression and its relationship with angiogenesis during experimental fibrosis. METHODS: Cirrhosis was induced in male Wistar rats by intraperitoneal administration of diethyl nitrosamine (DEN). The serial sections from liver tissues were stained with anti-CD34 and anti-TSP-1 antibodies before being quantitated by light microscopy. RESULTS: Our results showed that of TSP-1 expression gradually increases according to the severity of fibrosis (P < 0.05). Moreover, TSP-1 expression was found to be correlated with angiogenesis (P < 0.05). CONCLUSION: The correlative evidence of the link between TSP-1 and fibrosis or angiogenesis provided by this study suggests that besides its role as a strong promoter of transforming growth factor-beta 1 (TGF-beta 1), TSP-1 might have an additional role in liver fibrogenesis by stimulating angiogenesis and this protein could be a potential target to prevent fibrogenesis in chronic inflammatory diseases of the liver. PMID- 18407597 TI - Sustained virological response based on rapid virological response in genotype-3 chronic hepatitis C treated with standard interferon in the Pakistani population. AB - AIM: To document the sustained virological response (SVR) in rapid virological responders (RVR) of genotype-3 chronic hepatitis C with standard interferon (SdIF). METHODS: Hepatitis C genotype-3 patients during the period July 2006 and June 2007 were included. Complete blood counts, prothrombin time, ALT, albumin, qualitative HCV RNA were done. SdIF and ribavirin were given for 4 wk and qualitative HCV RNA was repeated. Those testing negative were allocated to group A while the rest were allocated to group-B. Treatment was continued a total of 16 and 24 wk for group A and B respectively. HCV RNA was repeated after 24 wk of treatment. End virological and sustained virological responses were compared by c2 test. ROC of pretreatment age, ALT and albumin were plotted for failure to achieve SVR. RESULTS: Of 74 patients treated, RCV RNA after 16 wk of therapy became undetectable in 34 (45.9%) and was detectable in 40 (54.1%) and were allocated to groups A and B respectively. SVR was achieved in 58.8% and 27.8% in groups A and B respectively. SVR rates were significantly higher in patients who had RVR as compared to those who did not (P = 0.0; gamma = 2). Both groups combined ETR and SVR were 70% and 33% respectively. ROC plots of pretreatment age, ALT and albumin for SVR showed only ALT to have a significantly large area under the curve. CONCLUSION: SVR rates were higher in patients who had RVR with SdIF and high pre treatment ALT values correlated to probability of having RVR. PMID- 18407598 TI - Cost saving by reloading the multiband ligator in endoscopic esophageal variceal ligation: a proposal for developing countries. AB - AIM: To assess the cost savings of reloading the multiband ligator in endoscopic esophageal variceal ligation (EVL) used on the same patient for subsequent sessions. METHODS: This single centre retrospective descriptive study analysed patients undergoing variceal ligation at a tertiary care centre between 1st January, 2003 and 30th June, 2006. The multiband ligator was reloaded with six hemorrhoidal bands using hemorrhoidal ligator for the second and subsequent sessions. Analysis of cost saving was done for the number of follow-up sessions for the variceal eradication. RESULTS: A total of 261 patients underwent at least one session of endoscopic esophageal variceal ligation between January 2003 and June 2006. Out of 261, 108 patients (males 67) agreed to follow the eradication program and underwent repeated sessions. A total of 304 sessions was performed with 2.81 sessions per patient on average. Thirty-two patients could not complete the program. In 76 patients (70%), variceal obliteration was achieved. The ratio of the costs for the session with reloaded ligator versus a session with a new ligator was 1:2.37. Among the patients who completed esophageal varices eradication, cost saving with reloaded ligator was 58%. CONCLUSION: EVL using reloaded multiband ligators for the follow-up sessions on patients undergoing variceal eradication is a cost saving procedure. Reloading the ligator thus is recommended especially for developing countries where most of the patients are not health insured. PMID- 18407599 TI - Effect of thermal cutaneous stimulation on the gastric motor activity: study of the mechanism of action. AB - AIM: To investigate the mechanism of action of thermal cutaneous stimulation on the gastric motor inhibition. METHODS: The gastric tone of 33 healthy volunteers (20 men, mean age 36.7 +/- 8.4 years) was assessed by a barostat system consisting of a balloon-ended tube connected to a strain gauge and air-injection system. The tube was introduced into the stomach and the balloon was inflated with 300 mL of air. The skin temperature was elevated in increments of 3 degree up to 49 degree and the gastric tone was simultaneously assessed by recording the balloon volume variations expressed as the percentage change from the baseline volume. The test was repeated after separate anesthetization of the skin and stomach with lidocaine and after using normal saline instead of lidocaine. RESULTS: Thermal cutaneous stimulation resulted in a significant decrease of gastric tone 61.2% +/- 10.3% of the mean baseline volume. Mean latency was 25.6 +/- 1.2 ms. After 20 min of individual anesthetization of the skin and stomach, thermal cutaneous stimulation produced no significant change in gastric tone. CONCLUSION: Decrease in the gastric tone in response to thermal cutaneous stimulation suggests a reflex relationship which was absent on individual anesthetization of the 2 possible arms of the reflex arc: the skin and the stomach. We call this relationship the "cutaneo-gastric inhibitory reflex". This reflex may have the potential to serve as an investigative tool in the diagnosis of gastric motor disorders, provided further studies are performed in this respect. PMID- 18407600 TI - DNMT3B 579 G>T promoter polymorphism and risk of esophagus carcinoma in Chinese. AB - AIM: To investigate the relationship between 579 G>T polymorphisms in the DNMT3B gene, which is involved in de novo methylation and associated with the risk of esophagus cancer (EC) in Chinese. METHODS: DNMT3B 579 G>T genotypes were determined by PCR-RFLP in 194 EC patients and 210 healthy controls matched for age and sex, who did not receive radiotherapy or chemotherapy for newly diagnosed and histopathologically confirmed EC. RESULTS: In control subjects, the frequency of T/T and G/T genotypes, and T and G alleles was 81.4%, 18.1%, 90.05% and 9.55%, respectively. The distribution of genotypes and allelotypes in the EC patients was not significantly different from that in the controls. When stratified by sex and age, there was still no significant association between the risks of EC and GT and GG genotypes. This study also showed a distinct difference in the distribution of DNMT3B and single nucleotide polymorphism (SNP) between Chinese and Koreans. CONCLUSION: DNMT3B 579 G>T polymorphism may not be a stratification marker to predict the susceptibility to EC, at least in Chinese. DNMT3B promoter SNP is diverse in ethnic populations. PMID- 18407601 TI - Anti-sense oligonucleotide labeled with technetium-99m using hydrazinonictinamide derivative and N-hydroxysuccinimidyl S-acetylmercaptoacetyltriglycline: a comparison of radiochemical behaviors and biological properties. AB - AIM: To explore and compare the radiochemical behavior and biological property of anti-sense oligonuc-leotide (ASON) labeled with technetium-99m using N hydroxysuccinimidyl S-acetylmercaptoacetyltriglycline (NHS-MAG(3)) and hydrazinonictinamide derivative (HYNIC). METHODS: After HYNIC and NHS-MAG(3) were synthesized, ASON was labeled with technetium-99m using HYNIC and NHS-MAG(3) as a bifunctional chelator. The in vivo and in vitro stability, binding rates of labeled compounds to serum albumen, biodistribution of (99m)Tc-MAG(3)-ASON and (99m)Tc-HYNIC-ASON in BALB/C mouse and its HT29 tumor cellular uptake were compared. RESULTS: The labeling efficiency and stability of (99m)Tc-MAG(3)-ASON were significantly higher than those of (99m)Tc-HYNIC-ASON (P = 0.02, and P = 0.03, respectively). (99m)Tc-MAG(3)-ASON had a significantly lower rate of binding to serum albumen than (99m)Tc-HYNIC-ASON (P < 0.05). In contrast to (99m)Tc-HYNIC-ASON, the biodistribution of (99m)Tc-MAG(3)-ASON was significantly lower in blood, heart, liver and stomach (P < 0.05), slightly lower in intestines and spleen (P > 0.05) and significantly higher in lung and kidney (P < 0.05). The HT29 tumor cellular uptake rate of (99m)Tc-MAG(3)-ASON was significantly higher than that of (99m)Tc-HYNIC-ASON (P < 0.05). CONCLUSION: (99m)Tc-MAG(3)-ASON shows superior radiochemical behaviors and biological properties than (99m)Tc-HYNIC ASON. (99m)Tc-MAG(3)-ASON is a potential radiopharmaceutical agent for in vivo application. PMID- 18407602 TI - Effects of simulated carbon dioxide and helium peumoperitoneum on proliferation and apoptosis of gastric cancer cells. AB - AIM: To investigate the effects of carbon dioxide (CO(2)) and helium insufflation administered at different pressures on the growth and apoptosis of cultured human gastric cancer cells. METHODS: The gastric cancer cells MKN-45 were exposed to a CO(2) and helium environment maintained at different pressures (0, 5, 10 and 15 mmHg). The cells were exposed to simulated pneumoperitoneum environment for 4 h, and pH of the culture media was measured after it was moved to normal conditions for 0, 2, 4, 6 and 8 h. Proliferation viability of MKN-45 was examined by 3 [4,5Dimethylthiazol-2-yl],5-diphenyltetrazolium bromide or triazolyl blue (MTT) assay after it was moved to normal conditions. Apoptotic ratio was measured by Annexin V-FITC/PI double labelled staining. RESULTS: The pH of media was acid and recovered to normal after 4 h in the CO(2) group while it was basic in the helium group. There was no difference between CO(2) groups (under 10 mmHg ) and control group (P > 0.05) in the proliferative viability of the cells. The cultured cells exposed to 15 mmHg CO(2) environment grew more slowly than control group from 4 to 7 d (P < 0.01 ) while there was no difference from 1 to 3 d (P > 0.05). The proliferative viability in helium group was not obviously different from the control group (P > 0.05). The apoptotic ratio of the cultured cells was markedly higher than that of the control group (P < 0.01) at 10 and 15 mmHg CO(2) insufflation pressure. In helium group, the apoptotic ratio was not obviously different from the control group (P > 0.05). CONCLUSION: There is no obvious effect in the proliferation and apoptosis of MKN-45 cells under 10 mmHg CO(2) insufflation pressure and helium in any pressure. Fifteen mmHg CO(2) insufflation pressure can inhibit the proliferation of the cells and improve apoptosis. PMID- 18407603 TI - Dynamic changes of IL-2/IL-10, sFas and expression of Fas in intestinal mucosa in rats with acute necrotizing pancreatitis. AB - AIM: To investigate dynamic changes of serum IL-2, IL-10, IL-2/IL-10 and sFas in rats with acute necrotizing pancreatitis. To explore the expression of Fas in intestinal mucosa of rats with acute necrotizing pancreatitis (ANP). METHODS: A total of 64 Sprague-Dawley (SD) rats were randomly divided into two groups: normal control group (C group), ANP group (P group). An ANP model was induced by injection of 50 g/L sodium taurocholate under the pancreatic membrane. Normal control group received isovolumetric injection of 9 g/L physiological saline solution using the same method. The blood samples of the rats in each group were obtained via superior mesenteric vein to measure levels of IL-2, IL-10, sFas and calculate the value of IL-2/IL-10. The levels of IL-2, IL-10 and sFas were determined by ELISA. The severity of intestinal mucosal injury was evaluated by pathologic score. The expression of Fas in intestinal mucosal tissue was determined by immunohistochemistry staining. RESULTS: Levels of serum IL-2 were significantly higher in P group than those of C group (2.79 +/- 0.51 vs 3.53 +/- 0.62, 2.93 +/- 0.89 vs 4.35 +/- 1.11, 4.81 +/- 1.23 vs 6.94 +/- 1.55 and 3.41 +/- 0.72 vs 4.80 +/- 1.10, respectively, P < 0.01, for all) and its reached peak at 6 h. Levels of serum IL-10 were significantly higher in P group than those of C group at 6 h and 12 h (54.61 +/- 15.81 vs 47.34 +/- 14.62, 141.15 +/- 40.21 vs 156.12 +/- 43.10, 89.18 +/- 32.52 vs 494.98 +/- 11.23 and 77.15 +/- 22.60 vs 93.28 +/- 25.81, respectively, P < 0.01, for all). The values of IL-2/IL-10 were higher significantly in P group than those of C group at 0.5 h and 2 h (0.05 +/- 0.01 vs 0.07 +/- 0.02 and 0.02 +/- 0.01 vs 0.03 +/- 0.01, respectively, P < 0.01, for all), and it were significantly lower than those of C group at 6 h (0.05 +/- 0.02 vs 0.01 +/- 0.01, P < 0.01) and returned to the control level at 12 h (0.04 +/- 0.01 vs 0.05 +/- 0.02, P > 0.05). In sFas assay, there was no significant difference between P group and C group (3.16 +/- 0.75 vs 3.31 +/- 0.80, 4.05 +/- 1.08 vs 4.32 +/- 1.11, 5.93 +/- 1.52 vs 5.41 +/- 1.47 and 4.62 +/- 1.23 vs 4.44 +/- 1.16, respectively, P > 0.05, for all). Comparison of P group and C group, the pathological changes were aggravated significantly in P group. Immunohistochemistry staining show the expression of Fas was absent in normal intestinal tissues, however, it gradually increased after induction of pancreatitis in intestinal tissue, then reached their peaks at 12 h. CONCLUSION: Fas were involved in the pathogenesis of pancreatitis associated intestinal injury. The mechanisms of Fas may be associated to Fas mediated T helper cell apoptosis. PMID- 18407604 TI - Clinicopathologic characteristics of intrahepatic cholangiocarcinoma in patients with positive serum a-fetoprotein. AB - AIM: To explore clinicopathologic characteristics of intrahepatic cholangiocarcinoma (ICC) in patients with positive serum a-fetoprotein (AFP). METHODS: One hundred and thirty one patients who underwent surgical dissection for pathologically confirmed ICC were divided into a positive AFP (> 20 ng/mL) group (n = 32) and a negative AFP group (n = 99), whose clinicopathologic features were analyzed and compared. RESULTS: The positive rate of HBsAg and liver cirrhosis of the positive AFP group was higher than that of the negative AFP group, while the positive rate of CA19-9 (> 37 U/mL) and the lymph node metastasis rate was lower. CONCLUSION: ICC patients with positive AFP share many clinicopathologic similarities with hepatocellular carcinoma. PMID- 18407605 TI - Risk factors for alcohol-related liver injury in the island population of China: a population-based case-control study. AB - AIM: To investigate the association of alcohol dose, duration of drinking and obesity with abnormal alcohol-related liver injury indicators, the prevalence of alcohol-related liver injury in the island population of China. METHODS: Randomized multistage stratified cluster sampling from the island population of China was used in the population-based case-control study. Then interview, physical examination, laboratory assessments and ultrasonography were done. RESULTS: Daily alcohol intake > or = 20 g, duration of drinking > or = 5 years and obesity were closely related to alcohol-related liver injury (P < 0.05). The odds-ratio (OR) (95% CI) was 1.965 (1.122-3.442), 3.412 (1.789-6.507) and 1.887 (1.261-2.824), respectively. The prevalence rate of alcohol-related liver injury in > or = 20 g daily alcohol intake group and < 20 g daily alcohol intake group was 37.14% and 12.06%, respectively. The prevalence rate of alcohol-related liver injury in > or = 5 years drinking group and < 5 years drinking group was 34.44% and 8.53%, respectively. No significant dose-response relation was found between daily alcohol intake and abnormal alcohol-related liver injury indicators as well as between duration of drinking and abnormal alcohol-related liver injury indicators. There was no significant difference in the prevalence of alcohol related liver injury between beer drinking group and yellow rice wine drinking group, hard liquor drinking group, multiple drinking group. CONCLUSION: The risk threshold of daily alcohol intake is 20 g and duration of drinking inducing alcohol-related liver injury 5 years in the island population of China. Liver injury induced by obesity should be concerned. PMID- 18407606 TI - Endoscopic diagnosis of gastrointestinal graft-versus-host disease. AB - AIM: To evaluate the diagnostic value of endoscopy in patients with gastrointestinal graft-versus-host disease (GI GVHD). METHODS: We identified 8 patients with GI GVHD following allogeneic hematopoietic stem cell trans plantation (HSCT). GVHD was defined histologically as the presence of gland apoptosis, not explained by other inflammatory or infectious etiologies. RESULTS: The symptoms of GI GVHD included anorexia, nausea, vomiting, watery diarrhea, abdominal pain, GI bleeding, etc. Upper endoscopic appearance varied from subtle mucosal edema, hyperemia, erythema to obvious erosion. Colonoscopic examination showed diffuse edema, hyperemia, patchy erosion, scattered ulcer, sloughing and active bleeding. Histological changes in GI GVHD included apoptosis of crypt epithelial cells, dropout of crypts, and lymphocytic infiltration in epithelium and lamina propria. The involvement of stomach and rectocolon varied from diffuse to focal. CONCLUSION: Endoscopy may play a significant role in early diagnosis of GI GVHD patients following allogeneic HSCT, and histologic examination of gastrointestinal biopsies is needed to confirm the final diagnosis. PMID- 18407607 TI - Extraintestinal heterotopic gastric tissue simulating acute appendicitis. AB - We describe the case of a 68 year old otherwise healthy male who presented to our emergency room with signs and symptoms of acute appendicitis. Exploratory surgery revealed a normal appendix. Further examination revealed an enlarged lymph node like mass of tissue near the appendix, in the ileocecal mesentery. This mass was removed and was found to be inflamed heterotopic gastric tissue. Although reports of heterotopic gastric tissue in the literature are common, we believe that this case represents the first report of inflamed heterotopic gastric tissue simulating appendicitis. PMID- 18407608 TI - Steroid responsive eosinophilic gastric outlet obstruction in a child. AB - Gastric outlet obstruction is a rare complication of eosinophilic gastroenteritis, most commonly treated surgically. We report a case of eosinophilic gastric outlet obstruction in a child that responded to conservative medical management. A brief review of this clinical entity is also provided. PMID- 18407609 TI - Ischemic colitis due to obstruction of mesenteric and splenic veins: a case report. AB - Ischemic injury to the bowel is a well known disease entity that has a wide spectrum of pathological and clinical findings. A sudden drop in the colonic blood supply is essential to its development. We encountered a 41-year-old male patient, who presented with abdominal pain and bloody diarrhea. A colonoscopy showed markedly edematous mucosa with tortuous dilatation of the veins and a deep ulceration at the rectosigmoid junction. On an abdominal computed tomography (CT) scan and CT angiography, the mesenteric and splenic veins were absent with numerous venous collaterals for drainage. The patient gradually responded to oral aminosalicylate therapy, and was in remission after nine months. In most cases, non-occlusive ischemic injury is caused by idiopathic form and occlusive ischemia is caused by abnormalities of arteries and acute venous thrombosis. However, chronic venous insufficiency due to obstruction of macrovascular mesenteric vein rarely causes ischemia of the bowel. This report describes the first case of ischemic colitis caused by obstruction of the mesenteric and splenic veins. PMID- 18407610 TI - Successful endoscopic treatment of biliary stricture following mesenteric tear caused by blunt abdominal trauma. AB - Biliary duct injuries are frequently iatrogenic, being associated with surgery for gallbladder stones. However, blunt abdominal trauma such as a motor vehicle crash is a rare cause of extrahepatic biliary stricture. A few reports have been published on biliary strictures treated with endoscopic therapy. In the present study, we describe a suprapancreatic biliary stricture associated with mesenteric tear following road traffic accident. We performed endoscopic stent placement, which was successful in relieving the biliary stricture. PMID- 18407611 TI - Appendiceal mucocele: case reports and review of current literature. AB - The mucocele of the appendix is an uncommon disorder which is often asymptomatic but sometimes causes acute appendicitis-like symptoms. Sometimes, patients with mucocele can present with confusing symptoms. Preoperative suspicion and diagnosis of appendiceal mucocele are important. Ultrasonography and computed tomography are useful tools for the diagnosis of appendiceal mucocele. It may be also recognised by colonoscopy as a smooth submucosal lesion of the cecum. Optimal management of the mucocele could be achieved through accurate preoperative diagnosis. Preoperative diagnosis is a major component for minimizing intra-operative and post-operative complications. We herein report five cases and discuss the diagnostic methods and surgical treatment. PMID- 18407612 TI - Tuberculous abscess in hepatoduodenal ligament: evaluation with contrast-enhanced computed tomography. AB - Two patients with tuberculous abscess in the hepatoduodenal ligament were studied. Both patients underwent contrast-enhanced computed tomography (CT) scan. The abscess showed a low density with an irregular thick wall in the hepatoduodenal ligament on CT images, the margin was poorly defined. Contrast enhanced CT images showed the contrast-enhanced thick wall, homogeneous and peripheral-enhanced lymph nodes. Although features of the tuberculous abscess in the hepatoduodenal ligament could be conspicuously shown with contrast-enhanced CT, further experience is needed to evaluate the potential value of CT in detecting early tuberculous abscess in relation to other entities in the hepatoduodenal ligament. PMID- 18407613 TI - Occult hepatitis C virus infection is more common than hepatitis B infection in maintenance hemodialysis patients. AB - Patients of end stage renal disease on maintenance hemodialysis were enrolled to study the prevalence of occult and dual hepatitis B virus (HBV) and hepatitis C virus (HCV) infection and non-occult hepatitis B and C virus infection. One hundred and two patients were enrolled. Thirty patients had HCV infection, three of them were positive in anti-HCV. So, 27 (90%) of HCV-positive patients had occult HCV infection. Eleven (11%) patients had HBV infection. Five patients were positive in anti-HBc or HBV-DNA, but negative in HBsAg (occult HBV infection). Three (3%) patients had dual HBV and HCV infection. None of the patients showed changes in viral markers during the follow-up of 8 mo on average (1-12 mo). PMID- 18407614 TI - Correlation of calculated excited-state energies and experimental quantum yields of luminescent Tb(III) beta-diketonates. AB - Theoretical calculations employing time-dependent density functional theory (TDDFT) are used to characterize the excited states of Tb(III) beta-diketonate complexes. Calculated results are compared directly with experimental results that together show a correlation between relative quantum yields and the excited state energies that depend on the electronic properties of the p,p'-substituent group associated with the coordinating N-donor neutral ligand. It is found that changes in the electron donating nature of the neutral ligand structure lead to shifts in the lowest triplet energy level of the complex that consequently change the relative quantum yield. This work provides critical direction for the synthesis of high quantum yield terbium complexes. PMID- 18407615 TI - Discrimination between Bacillus species by impedance analysis of individual dielectrophoretically positioned spores. AB - We combine the use of dielectrophoretic positioning with electrical impedance measurements to detect and discriminate between individual bacterial spores on the basis of their electrical response. Using lithographically defined microelectrodes, we use dielectrophoresis to manipulate individual bacterial spores between the electrodes. The introduction of a single spore between the microelectrodes produces a significant change in electrical response that is species-dependent. When positioned between two electrodes and an AC voltage was applied, single spores caused current increases averaging 6.8 (+/-2.4) pA for Bacillus mycoides to 1.18 (+/-0.37) pA for Bacillus licheniformis. Using a mixture of spores of two different species, we demonstrate the ability to distinguish the species of individual spores in real time. This work demonstrates the feasibility of using impedance measurements for real-time detection and discrimination between different types of spores. PMID- 18407616 TI - Imaging of metal ion dissolution and electrodeposition by anodic stripping voltammetry-scanning electrochemical microscopy. AB - We have developed a new imaging method for scanning electrochemical microscopy (SECM) employing fast-scan anodic stripping voltammetry (ASV) to provide sensitive and selective imaging of multiple chemical species at interfaces immersed in solution. A rapid cyclic voltammetry scan (100 V/s) is used along with a short preconcentration time (300-750 ms) to allow images to be acquired in a normal SECM time frame. A Hg-Pt film electrode is developed having an equivalent Hg thickness of 40 nm that has good sensitivity at short preconcentration times and also retains thin-film behavior with high-speed voltammetric stripping. Fast-scan anodic stripping currents are shown to be linear for 1-100 microM of Pb (2+) and Cd (2+) solutions using a preconcentration time of 300 ms. SECM images showing the presence of Pb (2+) and Cd (2+) at concentrations as low as 1 microM are presented. In addition, a single ASV-SECM image is shown to produce unique concentration maps indicating Cd (2+) and Pb (2+), generated in situ from a corroding sample, while simultaneously detecting the depletion of O 2 at this sample. The transient voltammetric response at the film electrode is simulated and shows good agreement with the experimental behavior. We discuss the behavior of images and concentration profiles obtained with different imaging conditions and show that mass-transport limitations in the tip-substrate gap can induce dissolution. ASV-SECM can thus be used to detect and study induced dissolution not only at bulk metal surfaces but also on underpotential deposition layers, in this case Cd and Pb on Pt. In addition, we discuss how surface diffusion phenomena may relate to the observed ASV-SECM behavior. PMID- 18407617 TI - Simple telemedicine for developing regions: camera phones and paper-based microfluidic devices for real-time, off-site diagnosis. AB - This article describes a prototype system for quantifying bioassays and for exchanging the results of the assays digitally with physicians located off-site. The system uses paper-based microfluidic devices for running multiple assays simultaneously, camera phones or portable scanners for digitizing the intensity of color associated with each colorimetric assay, and established communications infrastructure for transferring the digital information from the assay site to an off-site laboratory for analysis by a trained medical professional; the diagnosis then can be returned directly to the healthcare provider in the field. The microfluidic devices were fabricated in paper using photolithography and were functionalized with reagents for colorimetric assays. The results of the assays were quantified by comparing the intensities of the color developed in each assay with those of calibration curves. An example of this system quantified clinically relevant concentrations of glucose and protein in artificial urine. The combination of patterned paper, a portable method for obtaining digital images, and a method for exchanging results of the assays with off-site diagnosticians offers new opportunities for inexpensive monitoring of health, especially in situations that require physicians to travel to patients (e.g., in the developing world, in emergency management, and during field operations by the military) to obtain diagnostic information that might be obtained more effectively by less valuable personnel. PMID- 18407618 TI - Effective cell capture with tetrapeptide-functionalized carbon nanotubes and dual signal amplification for cytosensing and evaluation of cell surface carbohydrate. AB - A novel electrochemical cytosensing strategy was designed based on the specific recognition of integrin receptors on a cell surface to arginine-glycine-aspartic acid-serine (RGDS)-functionalized single-walled carbon nanotubes (SWNTs). The covalent conjugation of the RGDS tetrapeptide to SWNTs was proved with Raman and FT-IR spectra. The conjugated RGDS showed a predominant ability to capture cells on the electrode surface by the specific combination of RGD domains with integrin receptors. With the use of BGC-823 human gastric carcinoma cells (BGC cells) as a model, the cell surface mannosyl groups could specifically bind with horseradish peroxidase labeled concanavalin A, producing an electrochemical cytosensor. On the basis of the dual signal amplification of SWNTs and enzymatic catalysis, the cytosensor could respond down to 620 cells mL (-1) of BGC cells with a linear calibration range from 1.0 x 10 (3) to 1.0 x 10 (7) cells mL (-1), showing very high sensitivity. The dual signal amplification could be further used to evaluate the mannosyl groups on the cell surface, and the mannosyl groups on a single living intact BGC cell were detected to correspond to 5.3 x 10 (7) molecules of mannose. This strategy presents a promising platform for highly sensitive cytosensing and convenient evaluation of surface carbohydrates on living cells. PMID- 18407620 TI - Fast and efficient proteolysis by microwave-assisted protein digestion using trypsin-immobilized magnetic silica microspheres. AB - A fast and efficient proteolysis approach of microwave-assisted protein digestion was developed by using trypsin-immobilized magnetic silica (MS) microspheres. In the work, immobilization of the enzyme onto MS microspheres was very simple and only through a one-step reaction with 3-glycidoxypropyltrimethoxysilane (GLYMO) which provides the epoxy group as a reactive spacer. Considering that the magnetic particles are excellent microwave absorbers, we developed a novel microwave-assisted digestion method based on the easily prepared trypsin immobilized MS microspheres. This novel digestion method combined the advantages of immobilized trypsin and the rapid-fashion of microwave-assisted digestion, which resulted in high digestion efficiency. BSA and myoglobin were used as model proteins to optimize the conditions of this method. Peptide fragments produced in 15 s could be confidently identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis. Equivalent or better digestion efficiency was observed comparing to current in-solution digestion. Besides, because of the unique magnetic responsivity, the immobilized trypsin can be isolated easily with the help of an external magnet and thus used repeatedly. High activity was obtained even after seven runs of the trypsin immobilized MS microspheres. To further verify its efficiency in proteome analysis, one reversed-phase liquid chromatography (RPLC) fraction of rat liver extract was applied. After 15 s incubation, 16 totally unique peptides corresponding to two proteins were identified. Finally, the rat liver sample was used to evaluate its worth for the application. With analysis by liquid chromatography-electrospray-tandem mass spectrometry (LC-ESI-MS/MS), comparable digestion efficiency was observed with typical in-solution digestion but the incubation time was largely shortened. This new microwave-assisted digestion method will hasten the application of the proteome technique to biomedical and clinical research. PMID- 18407621 TI - Measurement of the cell-substrate separation and the projected area of an individual adherent cell using electric cell-substrate impedance sensing. AB - This work presents an electrical technique called electric cell-substrate impedance sensing to measure the cell-substrate separation and the projected area of an individual adherent cell. Cell adhesion and cell spreading are fundamental processes of adherent cells. By recording changes in the cell-substrate separation, the projected area, or both properties with time, the dynamics of cell spreading and cell adhesion can be studied. The advantage of this electrical technique is that it enables a measurement of many individual cells simultaneously. This is a great benefit to the study of heterogeneity in cell populations. The research consisted of building a custom impedance sensing setup, designing an in vitro assay to record an impedance spectrum of an individual living cell, and developing a data analysis method to obtain two properties of the cell from curve-fitting of the impedance spectrum. The values of the cell substrate separation and the projected area of an individual cell were within the expected ranges and in agreement with those obtained from optical microscopy. PMID- 18407622 TI - Simultaneous determination of species-specific isotopic composition of Hg by gas chromatography coupled to multicollector ICPMS. AB - This work presents the simultaneous online determination of the isotopic composition of different Hg species in a single sample by the hyphenation of gas chromatography (GC) with multicollector-inductively coupled plasma mass spectrometry (MC-ICPMS). With the use of commercially available instrumentation, precise and accurate species-specific Hg isotope delta values (per mil deviation of the Hg isotope ratio in the sample relative to a reference standard) have been obtained online from consecutive GC transient signals. The use of isothermal temperature programs to extend the elution of the Hg species, the proper selection of the peak integration window, as well as the preconcentration of real samples are critical to provide optimal counting statistics. Also, isotope ratio drift during transient signal elution was overcome by introducing a mixed Hg(II) and MeHg standard bracketing scheme and expressing all results using the delta notation relative to SRM NIST-3133. Using the proposed methodology, we have obtained an external 2SD precision of 0.56 per thousand for delta (202)Hg that is more than 10 times smaller than the overall Hg stable isotope variation thus far observed in terrestrial samples. The measurement of species-specific Hg isotopic composition relative to SRM NIST-3133 has been validated versus two other analytical techniques, i.e., conventional nebulization (CN) of Hg(II) solution and cold vapor (CV) generation of Hg (0) vapor. A good agreement between the species-specific delta values obtained by the different techniques has been obtained in secondary fractionated reference standard (UM-Almaden) and environmental matrixes, i.e., BCR-CRM 464 (tuna fish) and IAEA-085 (human hair). The results show mass-dependent and mass-independent fractionation in environmental samples, i.e., mass-independent fractionation of odd isotopes (199)Hg and (201)Hg in tuna fish was observed. This methodology provides new possibilities for the future study of species-specific stable isotope geochemistry of Hg and other trace metals. PMID- 18407623 TI - Kinetics and mechanism of nucleophilic addition to nitric oxide: secondary amine diazeniumdiolation. AB - The kinetics of nitric oxide (NO) addition to dipropylamine and pyrrolidine are determined for a range of solvents and NO pressures. A new isobaric technique is utilized to determine the order of NO, and the kinetics are shown to conform to reactions which are either mono- or bimolecular in NO depending upon the conditions. The mechanism is interpreted in terms of competitive addition of amine to either NO or its dimer. PMID- 18407625 TI - Ethene insertion into vanadium hydride intermediates formed via vanadium atom reaction with water or ethene: a matrix isolation infrared spectroscopic study. AB - The reaction of V atoms with H2O and various concentrations of C2D4 in argon has been investigated by matrix isolation infrared (IR) spectroscopy. Both C2D6 and CD2H-CD2H are observed as the major products of a set of parallel processes involving hydrogenation of ethene where the formal source of hydrogen is either C2D4 or H2O. Portions of the IR spectrum of CD2H-CD2H isolated in an argon matrix are observed for the first time. For experiments involving low concentrations of C2D4, irradiation of the matrix with light of wavelengths >455 nm results in VH2 formation, with limited observation of ethene hydrogenation. The source of H2 is believed to be due to photoelimination of molecular hydrogen from HO-V-H species, during matrix deposition, with OV as an additional product. Recombination of OV with available H2 in the matrix is proposed as the source of OVH2 under low ethene conditions. No evidence for VD2 formation is observed under our conditions. At higher C2D4 concentrations, VH2 formation is suppressed, while products of ethene hydrogenation are maximized. A second process competing with H2 elimination in which HO-V-H reacts with C2D4 is proposed. Parallel reaction schemes involving V atom insertion into the O-H bonds of water or the photoinduced insertion of V atoms into the C-D bonds of C2D4 are proposed to account for the observed hydrogenation products. In each mechanism, insertion of C2D4 into the V-H or V-D bonds of transient intermediates is followed by photoinduced elimination of the associated ethane isotopomer. PMID- 18407626 TI - F5SN(H)Xe+; a rare example of xenon bonded to sp3-hybridized nitrogen; synthesis and structural characterization of [F5SN(H)Xe][AsF6]. AB - The salt [F5SN(H)Xe][AsF6] has been synthesized by the reaction of [F5SNH3][AsF6] with XeF2 in anhydrous HF (aHF) and BrF5 solvents and by solvolysis of [F3S triple bond NXeF][AsF6] in aHF. Both F5SN(H)Xe(+) and F5SNH3(+) have been characterized by (129)Xe, (19)F, and (1)H NMR spectroscopy in aHF (-20 degrees C) and BrF5 (supercooled to -70 degrees C). The yellow [F5SN(H)Xe][AsF6] salt was crystallized from aHF at -20 degrees C and characterized by Raman spectroscopy at -45 degrees C and by single-crystal X-ray diffraction at -173 degrees C. The Xe-N bond length (2.069(4) A) of the F5SN(H)Xe(+) cation is among the shortest Xe-N bonds presently known. The cation interacts with the AsF6(-) anion by means of a Xe---F-As bridge in which the Xe---F distance (2.634(3) A) is significantly less than the sum of the Xe and F van der Waals radii (3.63 A) and the AsF6(-) anion is significantly distorted from Oh symmetry. The (19)F and (129)Xe NMR spectra established that the [F5SN(H)Xe][AsF6] ion pair is dissociated in aHF and BrF5 solvents. The F5SN(H)Xe(+) cation decomposes by HF solvolysis to F5SNH3(+) and XeF2, followed by solvolysis of F5SNH3(+) to SF6 and NH4(+). A minor decomposition channel leads to small quantities of F5SNF2. The colorless salt, [F5SNH3][AsF6], was synthesized by the HF solvolysis of F3S triple bond NAsF5 and was crystallized from aHF at -35 degrees C. The salt was characterized by Raman spectroscopy at -160 degrees C, and its unit cell parameters were determined by low-temperature X-ray diffraction. Electronic structure calculations using MP2 and DFT methods were used to calculate the gas-phase geometries, charges, bond orders, and valencies as well as the vibrational frequencies of F 5SNH3(+) and F5SN(H)Xe(+) and to aid in the assignment of their experimental vibrational frequencies. In addition to F5TeN(H)Xe(+), the F5SN(H)Xe(+) cation provides the only other example of xenon bonded to an sp (3)-hybridized nitrogen center that has been synthesized and structurally characterized. These cations represent the strongest Xe-N bonds that are presently known. PMID- 18407624 TI - Characterization of the Fe site in iron-sulfur cluster-free hydrogenase (Hmd) and of a model compound via nuclear resonance vibrational spectroscopy (NRVS). AB - We have used (57)Fe nuclear resonance vibrational spectroscopy (NRVS) to study the iron site in the iron-sulfur cluster-free hydrogenase Hmd from the methanogenic archaeon Methanothermobacter marburgensis. The spectra have been interpreted by comparison with a cis-(CO)2-ligated Fe model compound, Fe(S2C2H4)(CO)2(PMe3)2, as well as by normal mode simulations of plausible active site structures. For this model complex, normal mode analyses both from an optimized Urey-Bradley force field and from complementary density functional theory (DFT) calculations produced consistent results. For Hmd, previous IR spectroscopic studies found strong CO stretching modes at 1944 and 2011 cm(-1), interpreted as evidence for cis-Fe(CO)2 ligation. The NRVS data provide further insight into the dynamics of the Fe site, revealing Fe-CO stretch and Fe-CO bend modes at 494, 562, 590, and 648 cm(-1), consistent with the proposed cis-Fe(CO)2 ligation. The NRVS also reveals a band assigned to Fe-S stretching motion at approximately 311 cm(-1) and another reproducible feature at approximately 380 cm(-1). The (57)Fe partial vibrational densities of states (PVDOS) for Hmd can be reasonably well simulated by a normal mode analysis based on a Urey-Bradley force field for a five-coordinate cis-(CO)2-ligated Fe site with additional cysteine, water, and pyridone cofactor ligands. A "truncated" model without a water ligand can also be used to match the NRVS data. A final interpretation of the Hmd NRVS data, including DFT analysis, awaits a three-dimensional structure for the active site. PMID- 18407627 TI - Photophysical behavior of open-shell first-row transition-metal octabutoxynaphthalocyanines: CoNc(OBu)8 and CuNc(OBu)8 as case studies. AB - Ultrafast photodynamics and density functional theory/time-dependent density functional theory (DFT/TDDFT) results for complexes of Co and Cu with 5,9,14,18,23,27,32,36-octabutoxynaphthalocyanine [CoNc(OBu)8 and CuNc(OBu)8] are reported. As a basis for this work, details concerning the syntheses of these complexes and the corresponding Zn complex (used as a reference) are given. Transient absorption spectrometry with femtosecond time resolution combined with a detailed DFT/TDDFT analysis has been employed to construct a complete picture of the excited-state dynamics after Q-band excitation of the Co and Cu complexes and to gain an understanding of the relationship between the nature of the metal center and the excited-state lifetime. The Co complex was shown to return to its ground state via two competing channels: a (2)T1(pi, pi*) state that decayed with a lifetime of 1 ps and a low-lying (2)(d, d) state that repopulated the ground state surface with a lifetime of 15 ps. CuNc(OBu)8 showed ground-state repopulation from the (2)T1(pi, pi*) state via a lower-lying ligand-to-metal charge-transfer (LMCT) state that was completed within a few nanoseconds. The photophysical behavior of the cobalt and copper complexes was compared to that previously reported for the nickel analog in an effort to highlight the effect of the central metal on the nature and rates of the deactivation pathways. The results described in this work provide basic knowledge that is relevant to the use of these compounds as photothermal sensitizers in cancer therapy. PMID- 18407628 TI - Ligand structure, conformational dynamics, and excited-state electron delocalization for control of photoinduced electron transfer rates in synthetic donor-bridge-acceptor systems. AB - Synthesis, ground-, and excited-state properties are reported for two new electron donor-bridge-acceptor (D-B-A) molecules and two new photophysical model complexes. The D-B-A molecules are [Ru(bpy)2(bpy-phi-MV)](PF6)4 (3) and [Ru(tmb)2(bpy-phi-MV)](PF6)4 (4), where bpy is 2,2'-bipyridine, tmb is 4,4',5,5' tetramethyl-2,2'-bipyridine, MV is methyl viologen, and phi is a phenylene spacer. Their model complexes are [Ru(bpy)2(p-tol-bpy)](PF6)2 (1) and [Ru(tmb)2(p tol-bpy)](PF6)2 (2), where p-tolyl-bpy is 4-(p-tolyl)-2,2'-bipyridine. Photophysical characterization of 1 and 2 indicates that 2.17 eV and 2.12 eV are stored in their respective (3)MLCT (metal-to-ligand charge transfer) excited state. These values along with electrochemical measurements show that photoinduced electron transfer (D*-B-A-->D (+)-B-A(-)) is favorable in 3 and 4 with DeltaG degrees(ET)=-0.52 eV and -0.62 eV, respectively. The driving force for the reverse process (D(+)-B-A(-) --> D-B-A) is also reported: DeltaG degrees(BET)=-1.7 eV for 3 and -1.5 eV for 4. Transient absorption (TA) spectra for 3 and 4 in 298 K acetonitrile provide evidence that reduced methyl viologen is observable at 50 ps following excitation. Detailed TA kinetics confirm this, and the data are fit to a model to determine both forward (k(ET)) and back (k(BET)) electron transfer rate constants: k(ET)=2.6 x 10(10) s(-1) for 3 and 2.8 x 10(10) s(-1) for 4; k(BET)=0.62 x 10(10) s(-1) for 3 and 1.37 x 10(10) s(-1) for 4. The similar rate constants k ET for 3 and 4 despite a 100 meV driving force (DeltaG degrees(ET)) increase suggests that forward electron transfer in these molecules in room temperature acetonitrile is nearly barrierless as predicted by the Marcus theory. The reduction in electron transfer reorganization energy necessary for this barrierless reactivity is attributed to excited-state electron delocalization in the (3)MLCT excited states of 3 and 4, an effect that is made possible by excited-state conformational changes in the aryl-substituted ligands of these complexes. PMID- 18407629 TI - Towards zirconium phosphonate-based microarrays for probing DNA-protein interactions: critical influence of the location of the probe anchoring groups. AB - Terminal phosphate groups on double-stranded DNA probes bind strongly to glass substrates coated with a zirconium phosphonate monolayer, and probes immobilized in this way as microarrays can be used to detect protein targets. The sensitivity of the microarray was shown to be enhanced by the use of a polyguanine segment ((G)n , n > or = 5) as a spacer between the phosphate linker and the protein interaction domain. More importantly, the presence of phosphate linkers on both ends of the dsDNA probes leads to significant enhancement of target capture. The relevant characteristics of the different probes when bound to the surface were determined, by the original use of a combination of surface characterization techniques (XPS, AFM, and Sarfus). In this context, the location of the phosphate linkers in the duplex probes was found to result in different probe surface coverage and presentation on the surface, which affect subsequent interactions with the target protein. PMID- 18407630 TI - Mapping the evolution of adsorption of water in nanoporous silica by in situ solid-state 1H NMR spectroscopy. AB - A recently developed technique for carrying out in situ solid-state NMR studies of adsorption processes in nanoporous materials is applied to map the evolution of the adsorption of water on the widely used siliceous nanoporous host material MCM-41. The technique allows the very earliest stages of the adsorption process to be probed, and using solid-state 1H NMR, the time resolution is of the order of a few hundred seconds. This work reveals that different water environments are populated at different stages of the adsorption process and reveals insights regarding both the sequence in which these different environments become populated and the exchange of water molecules between these environments. The results also provide access to information on the kinetics of the adsorption process, revealing an initial regime of rapid water adsorption up to ca. 1 wt % water, followed by a regime of slower water adsorption. In both regimes, the amount of water adsorbed increases linearly with time. As demonstrated by these results, the approach employed in this work creates new opportunities for investigating, in unprecedented detail, adsorption processes in nanoporous materials. PMID- 18407631 TI - Asymmetric droplet interface bilayers. AB - In cell membranes, the lipid compositions of the inner and outer leaflets differ. Therefore, a robust model system that enables single-channel electrical recording with asymmetric bilayers would be very useful. We and others recently developed the droplet interface bilayer (DIB), which is formed by connecting lipid monolayer-encased aqueous droplets submerged in an oil-lipid mixture. Here, we incorporate lipid vesicles of different compositions into aqueous droplets and immerse them in an oil bath to form asymmetric DIBs (a-DIBs). Both alpha-helical and beta-barrel membrane proteins insert readily into a-DIBs, and their activity can be measured by single-channel electrical recording. We show that the gating behavior of outer membrane protein G (OmpG) from Escherichia coli differs depending on the side of insertion in an asymmetric DIB with a positively charged leaflet opposing a negatively charged leaflet. The a-DIB system provides a general platform for studying the effects of bilayer leaflet composition on the behavior of ion channels and pores. PMID- 18407634 TI - 2-Fluoro-ATP as a versatile tool for 19F NMR-based activity screening. AB - 19F NMR-based methods have found utility in activity-based screening assays. However, because enzymes catalyze a diverse set of reactions, a large variety of fluorinated substrates would need to be identified to target each one separately. We have developed a more streamlined approach that is applicable to many enzymes that utilize ATP as a substrate. In this method, a fluorine-containing ATP analogue, 2-fluoro-ATP, is used to monitor the reaction. Applications are described for nicotinamide adenine dinucleotide synthetase and 3-phosphoinositide dependent kinase-1. Fragment screening results for the latter indicate that this technique can identify compounds that inhibit as well as activate reactions. The present results, together with previous biochemical studies from other laboratories, have shown that 2-fluoro-ATP can serve as a substrate for nine enzymes that are representative of three of the six enzyme subclasses, namely the transferases, hydrolases, and ligases. This suggests that 2-fluoro-ATP is suitable as a universal tool for screening ATP-requiring enzymes. Importantly, 2 fluoro-ATP has been determined to be a valid substrate for a variety of kinases, including both small molecule and protein kinases, suggesting that it may be useful for investigating the large number of pharmaceutically relevant kinases. PMID- 18407635 TI - A new method for N-N bond cleavage of N,N-disubstituted hydrazines to secondary amines and direct ortho amination of naphthol and its analogues. AB - An unexpected reaction of N,N-disubstituted hydrazine with naphthol and its analogues under simply thermal conditions has been disclosed. 2-Naphthol reacted with various N,N-disubstituted hydrazines under argon to afford 1-amino-2 naphthol and the corresponding secondary amines in excellent yields. Ortho amination of 2-naphthols, hydroxyquinoline, and naphthalenamine occurred when they reacted with N-methyl-N-phenylhydrazine. PMID- 18407636 TI - Synthesis of two mirror image 4-helix junctions derived from glycerol nucleic acid. AB - Structural DNA nanotechnology relies on Watson-Crick base pairing rules to assemble DNA motifs into diverse arrangements of geometric shapes and patterns. While substantial effort has been devoted to expanding the programmability of natural DNA, considerably less attention has been given to the development of nucleic acid structures based on non-natural DNA polymers. Here we describe the use of glycerol nucleic acid (GNA), a simple polymer based on an acyclic repeating unit, as an alternative genetic material for assembling nucleic acid nanostructures independent of RNA or DNA recognition. We synthesized two 4-helix junctions based entirely on GNA self-pairing and showed that GNA provides easy access to highly stable nanostructures with left- and right-handed helical configurations. PMID- 18407637 TI - Total synthesis of lobatoside E, a potent antitumor cyclic triterpene saponin. AB - Lobatoside E, a novel and complex cyclic triterpene saponin showing potent antitumor activities, has been synthesized for the first time, employing a highly modular approach. The synthesis, starting with oleanolic acid, D-glucose, D galactose, L-arabinose, and L-rhamnose, requires a total of 73 steps, with the longest linear sequence of 31 steps and in 1.2% overall yield. PMID- 18407638 TI - Gadolinium chelate coated gold nanoparticles as contrast agents for both X-ray computed tomography and magnetic resonance imaging. AB - Functionalized gold nanoparticles were applied as contrast agents for both in vivo X-ray and magnetic resonance imaging. These particles were obtained by encapsulating gold cores within a multilayered organic shell which is composed of gadolinium chelates bound to each other through disulfide bonds. The contrast enhancement in MRI stems from the presence of gadolinium ions which are entrapped in the organic shell, whereas the gold core provides a strong X-ray absorption. This study revealed that these particles suited for dual modality imaging freely circulate in the blood vessels without undesirable accumulation in the lungs, spleen, and liver. PMID- 18407639 TI - Correlating the crystal structure of a thiol-protected Au25 cluster and optical properties. AB - The total structure determination of thiol-protected Au clusters has long been a major issue in cluster research. Herein, we report an unusual single crystal structure of a 25-gold-atom cluster (1.27 nm diameter, surface-to-surface distance) protected by eighteen phenylethanethiol ligands. The Au25 cluster features a centered icosahedral Au13 core capped by twelve gold atoms that are situated in six pairs around the three mutually perpendicular 2-fold axes of the icosahedron. The thiolate ligands bind to the Au25 core in an exclusive bridging mode. This highly symmetric structure is distinctly different from recent predictions of density functional theory, and it also violates the empirical golden rule "cluster of clusters", which would predict a biicosahedral structure via vertex sharing of two icosahedral M13 building blocks as previously established in various 25-atom metal clusters protected by phosphine ligands. These results point to the importance of the ligand-gold core interactions. The Au25(SR)18 clusters exhibit multiple molecular-like absorption bands, and we find the results are in good correspondence with time-dependent density functional theory calculations for the observed structure. PMID- 18407641 TI - A small neutral molecule with two noble-gas atoms: HXeOXeH. AB - A novel noble-gas compound, HXeOXeH, is identified using IR spectroscopy, and it seems to be the smallest known neutral molecule with two noble-gas atoms. HXeOXeH is prepared using, for example, UV photolysis of water in solid xenon and subsequent annealing at 40-45 K. The experimental observations are fully supported by extensive quantum chemical calculations. A large energy release of 8.3 eV is computationally predicted for the decomposition of HXeOXeH into the 2Xe + H2O global energy minimum. HXeOXeH may represent a first step toward the possible preparation of (Xe-O)n chains and it may be relevant to the terrestrial "missing xenon" problem. PMID- 18407640 TI - Fluidic and air-stable supported lipid bilayer and cell-mimicking microarrays. AB - As drug delivery, therapy, and medical imaging are becoming increasingly cell specific, there is a critical need for high fidelity and high-throughput screening methods for cell surface interactions. Cell membrane-mimicking surfaces, i.e., supported lipid bilayers (SLBs), are currently not sufficiently robust to meet this need. Here we describe a method of forming fluidic and air stable SLBs through tethered and dispersed cholesterol groups incorporated into the bottom leaflet. Achieving air stability allows us to easily fabricate SLB microarrays from direct robotic spotting of vesicle solutions. We demonstrate their application as cell membrane-mimicking microarrays by reconstituting peripheral as well as integral membrane components that can be recognized by their respective targets. These demonstrations establish the viability of the fluidic and air-stable SLB platform for generating content microarrays in high throughput studies, e.g., the screening of drugs and nanomedicine targeting cell surface receptors. PMID- 18407642 TI - Organocatalytic asymmetric transfer hydrogenation in aqueous media using resin supported peptide having a polyleucine tether. AB - A resin-supported N-terminal prolyl peptide having a beta-turn motif and hydrophobic polyleucine chain effectively catalyzed the asymmetric transfer hydrogenation under aqueous conditions. The polyleucine tether provides a hydrophobic cavity in aqueous media that brought about a remarkable acceleration of the reaction. In addition, the polyleucine chain also turned out to be essential for high enantioselectivity. PMID- 18407643 TI - Diastereoselective synthesis of enantiopure acyclic beta,beta'-disubstituted vinylsulfoxides. AB - Thermal elimination of sulfenic acid from enantiopure beta,beta'-disubstituted bis-sulfoxides allows the stereoselective synthesis of enantiopure acyclic beta,beta'-disubstituted vinylsulfoxides. This mild and stereospecific synthesis provides either ( E) or ( Z) vinylsulfoxides in high yields and is compatible with acid or base sensitive functional groups. PMID- 18407644 TI - Solid-phase synthesis of azidomethylene inhibitors targeting cysteine proteases. AB - An efficient strategy for the solid-phase synthesis of azidomethylene inhibitors targeting cysteine proteases is described. The method is highlighted by its compatibility with readily available building blocks, as well as its ability to accommodate different functional groups. A 249-member library has thus far been successfully synthesized, characterized, and screened against Caspase-1, -3 and 7. PMID- 18407645 TI - New, general, and practical preparation of methyl ketones via the direct coupling of amides with CH2Cl2 promoted by TiCl4/Mg. AB - The direct coupling of a variety of amides with CH2 Cl2 or CD2 Cl2 promoted by TiCl 4/Mg/THF provides an extremely simple, practical, selective, and efficient approach for the construction of methyl ketones. The efficiency and practicability of this chemistry is illustrated by the very simple synthesis of deuterated methyl ketones. PMID- 18407646 TI - Phorbasins D-F: diterpenyl-taurines from a southern Australian marine sponge, Phorbas sp. AB - Fractionation of an Australian marine sponge, Phorbas sp., yielded the known phorbasins B (1) and C (2) together with three novel diterpenyl-taurines, phorbasins D-F (3-5). Partial absolute structures were assigned on the basis of detailed spectroscopic analysis. Phorbasins D-F (3-5) are the first reported examples of terpenyl-taurines linked via an amine moiety, and phorbasins E and F (4 and 5) incorporate an unprecedented heterocycle. A plausible biosynthetic route is proposed for the incorporation of taurine and heterocycle formation. PMID- 18407647 TI - Introduction of functional groups into peptides via N-alkylation. AB - An optimized protocol for the mild and selective Fukuyama-Mitsunobu reaction was used for mono- and di- N-alkylation on solid support. Thereby, nonfunctionalized aliphatic and aromatic residues are quickly introduced into transiently protected, primary amines of a linear peptide. N-Alkylation can also be used to implement alkyl chains carrying (protected) functionalities suited for subsequent modification. Applicability of this method is demonstrated by various N-alkylated analogues of a cyclic CXCR4 receptor antagonist originally developed by Fujii et. al. PMID- 18407648 TI - LDA-mediated selective addition reaction of vinylidenecyclopropanes with aldehydes, ketones, and enones: facile synthesis of vinylcyclopropenes, allenols, and 1,3-enynes. AB - Highly selective addition reaction of vinylidenecyclopropanes 1 was realized by treatment with LDA in THF and quenching with aldehydes, ketones, and enones. A number of vinylcyclopropenes, allenols, and 1,3-enynes were obtained selectively in moderate to good yields depending on the nature of different electrophiles. PMID- 18407649 TI - Effect of apple particle state on the release of volatile compounds in a new artificial mouth device. AB - Varying the crushing parameters in a model mouth apparatus gave different crushed apple samples, which were compared to apples crushed in the human mouth by six people. An image analysis method was developed to measure the similarity between apple particles after crushing in the artificial mouth and in the human mouth. Thus, experimental conditions were determined that produced fruit in a state closest to that obtained after mastication in a human mouth. The influence of these different conditions on the quantity of released volatile compounds was then studied. PMID- 18407650 TI - Kinetic modeling of formic acid pulping of bagasse. AB - Organic solvent or organosolv pulping processes are alternatives to soda or kraft pulping to delignify lignocellulosic materials for the production of paper pulp. Formic acid, a typical organosolv system, has been presently examined under atmospheric pressure to pulp bagasse fibers. It was shown that efficient bagasse pulping was achieved when the formic acid concentration was limited to 90% (v/v). A statistical kinetic model based on the experimental results for the delignification of bagasse during formic acid pulping was developed that can be described as follows: D (delignification) = 0.747 x C(formicacid) (1.688) x (1 - e(-0.05171t)), an equation that can be used to predict the lignin content in formic acid during the pulping process. The delignification of bagasse by 90% formic acid was almost completed after approximately 80 min, while extended pulping did not improve the delignification but tended to degrade the carbohydrates in bagasse, especially the hemicelluloses, which were rapidly hydrolyzed at the onset of pulping. PMID- 18407651 TI - Chemical response of hardwood oligosaccharides as a statistical function of isolation protocol. AB - Hardwoods contain a substantial amount of hemicellulose-type oligosaccharides, the chemical structures of which are typically categorized in the xylan saccharide class. The efficient and selective isolation of these hemicelluloses is regarded as one of the most critical obstacles to overcome for their eventual biomaterial and bioenergy utilization. Therefore, the objective of the current work was to perform a fundamental exploration of the function of extraction pH, temperature, and time on the final chemical properties of the extracted hemicelluloses. The extraction chemistries employed varied pH and showed that acidic conditions provided a higher extractive yield versus alkaline conditions. An alkaline environment gave higher yields than an acidic environment and also resulted in much higher lignin removal from wood, especially at high temperature. In general, control of the pH chemistry leads to a change in the carboxylic acid groups present in the extracts. When the extraction pH changed from acidic to alkaline, the total carboxylic acid group content in the hemicelluloses removed significantly increased due to more acetic acid group hydrolysis and increased lignin degradation. PMID- 18407652 TI - Rapid determination of crocetin esters and picrocrocin from saffron spice (Crocus sativus L.) using UV-visible spectrophotometry for quality control. AB - The aim of this work was the development of multivariate models able to determine the content of the main crocetin esters and picrocrocin from spectrophotometric data that could be used for routine quality control of saffron. These compounds were determined with HPLC in Spanish saffron, and their absorbance spectra from 190 to 700 nm were simultaneously monitored. Partial least-squares regression (PLSR) models have been obtained and applied to the determination of individual crocetin esters, to the sum of crocetin esters, and to picrocrocin. A modification of the Kennard-Stone algorithm was used to divide the pool of samples into calibration and validation subsets. The best predictions were obtained with the sum of crocetin esters model, followed by the model for cis crocetin (beta- D-glucosyl)-(beta- D-gentiobiosyl) ester, trans-crocetin di-(beta D-gentiobiosyl) ester, and trans-crocetin (beta- D-glucosyl)-(beta- D gentiobiosyl) ester, whereas the worst predictions were found with the picrocrocin and trans-crocetin (beta- D-gentiobiosyl) ester models. These models may enhance quality control in saffron enterprises. PMID- 18407653 TI - A simple and selective method for the measurement of azadirachtin and related azadirachtoid levels in fruits and vegetables using liquid chromatography electrospray ionization tandem mass spectrometry. AB - Neem-based insecticides containing azadirachtin and related azadirachtoids are widely used in agriculture. Here, we report an analytical method for the rapid and accurate quantification of the insecticide azadirachtin A and B and other azadirachtoids such as salannin, nimbin, and their deacetylated analogues on tomatoes and peaches. Azadirachtoids were extracted from fruits and vegetables with acetonitrile. Using high-performance liquid chromatography/electrospray ionization tandem mass spectrometer, azadirachtoids were selectively detected monitoring the multiple reaction transitions of sodium adduct precursor ions. For azadirachtin A, calibration was linear over a working range of 1-1000 microg/L with r > 0.996. The limit of detection and limit of quantification for azadirachtin A were 0.4 and 0.8 microg/kg, respectively. The presence of interfering compounds in the peach and tomato extracts was evaluated and found to be minimal. Because of the linear behavior, it was concluded that the multiple reaction transitions of sodium adduct ions can be used for analytical purposes, that is, for the identification and quantification of azadirachtin A and B and related azadirachtoids in fruit and vegetable extracts at trace levels. PMID- 18407654 TI - Feasibility of diode-array instruments to carry near-infrared spectroscopy from laboratory to feed process control. AB - Near-infrared calibrations were developed for the instantaneous prediction of the chemical and ingredient composition of intact compound feeds. Two rather different instruments were compared (diode array vs grating monochromator). The grating monochromator was used in a static mode in the laboratory, whereas the diode-array instrumentbetter adapted to online analysiswas placed on a conveyor belt to simulate measurements at a feed mill plant. Modified partial least squares (MPLS) equations were developed using the same set of samples analyzed in the two instruments. Sample set 1 ( N = 398) was used to predict crude protein (CP) and crude fiber (CF), while sample set 2 ( N = 393) was used for the prediction of one macroingredient (sunflower meal, SFM) and one microingredient (mineral-vitamin premix, MVP). The standard error of cross-validation (SECV) and the coefficient of determination (R2) values for CF were better using the monochromator instrument. However, results obtained for CP, SFM, and MVP using the samples analyzed in the diode-array instrument showed similar or even greater accuracy than those obtained using samples analyzed in the grating monochromator. The excellent predictive ability [R2> 0.95; RPD (ratio of standard deviation to SECV) > 3] obtained for CP, CF, and SFM opens the way for the online use of NIRS diode-array instruments for surveillance and monitoring in the manufacture, processing, and marketing of compound feeds. R2, RPD, and SECV values for MVP showed similar performance for both instruments. Although RPD values did not reach the minimum recommended for quantitative analysis, results are encouraging for an ingredient present in feed compounds in such very low amounts. PMID- 18407655 TI - Influence of the addition of raspberry seed extract on changes in the volatile pattern of stored model breakfast cereal. AB - Laboratory-prepared muesli-type breakfast cereal (mixture of oat flakes, wheat flakes, corn flakes, hazelnuts, raisins, sunflower seeds, and flax seeds) was subjected to accelerated storage test at 60 degrees C with or without the addition of red raspberry seed extract. The oxidative changes in muesli resulting in the formation of secondary oxidation products were evaluated using solid phase microextraction (HS-SPME) and solvent-assisted flavor evaporation (SAFE) to isolate volatiles and GC-MS and chromatography-olfactometry to quantify them and determine the key odorants. During 14 days of storage the total amount of volatile compounds changed from 1.0 mg/kg, in freshly prepared muesli, to 32 mg/kg after storage. The predominant compound was hexanal; its content during storage increased 20-fold, reaching 17 mg/kg. Red raspberry seed extract addition limited the rate of lipid oxidation, and the total amount of volatiles was estimated at 11 mg/kg and that of hexanal at almost 5 mg/kg. An elevated temperature of the storage test also influenced the crucial flavor compounds determined using aroma extract dilution analysis (AEDA). The flavor dilution factor (FD) values for volatile lipid oxidation products were lower in samples with red raspberry seed extract added. PMID- 18407656 TI - Application of a modified Haug and Lantzsch method for the rapid and accurate photometrical phytate determination in soybean, wheat, and maize meals. AB - A modified version of the Haug and Lantzsch method for rapid photometrical phytate determination was applied for the analysis of phytate in soybean, wheat, and maize meals. In comparison to the original protocol, the amount of the toxic reagent thioglycolic acid is reduced substantially to minimize potential health risks for laboratory personnel. Different extraction conditions for soybean meal were tested, and boiling for at least 30 min was found to be necessary to remove an interfering compound in soybean meal extracts. Phytate contents determined according to the modified Haug and Lantzsch method did not differ from those obtained by measuring total precipitated phosphorus or by sensitive and specific high-performance ion chromatography. Applicability and accuracy of the modified method for phytate analysis in major feed substrates, including soy-based textured vegetable protein, were demonstrated. PMID- 18407657 TI - b6f-Associated chlorophyll: structural and dynamic contribution to the different cytochrome functions. AB - Cytochromes bc1/b6f complexes catalyze electron transfer from lipid- to water soluble carriers in both the respiratory and photosynthetic processes. They contain several common redox cofactors, while a chlorophyll a molecule, the function of which is still enigmatic, is only present in b b6f-type complexes. In this work, we describe a mutagenesis approach aimed at characterizing the role of this pigment. Mutants of the binding pocket were constructed to obtain cytochrome (cyt) b6f f complexes altered in chlorophyll position and/or stability. On the basis of a combined biochemical and functional analysis, we conclude that the chlorophyll plays a major structural role in the complex. Moreover, the chlorophyll and its binding pocket may also be implicated in the regulation of photosynthetic state transitions, a function that is specific to cyt b6f complexes. PMID- 18407658 TI - Probing the dynamic interface between trimethylamine dehydrogenase (TMADH) and electron transferring flavoprotein (ETF) in the TMADH-2ETF complex: role of the Arg-alpha237 (ETF) and Tyr-442 (TMADH) residue pair. AB - We have used multiple solution state techniques and crystallographic analysis to investigate the importance of a putative transient interaction formed between Arg alpha237 in electron transferring flavoprotein (ETF) and Tyr-442 in trimethylamine dehydrogenase (TMADH) in complex assembly, electron transfer, and structural imprinting of ETF by TMADH. We have isolated four mutant forms of ETF altered in the identity of the residue at position 237 (alphaR237A, alphaR237K, alphaR237C, and alphaR237E) and with each form studied electron transfer from TMADH to ETF, investigated the reduction potentials of the bound ETF cofactor, and analyzed complex formation. We show that mutation of Arg-alpha237 substantially destabilizes the semiquinone couple of the bound FAD and impedes electron transfer from TMADH to ETF. Crystallographic structures of the mutant ETF proteins indicate that mutation does not perturb the overall structure of ETF, but leads to disruption of an electrostatic network at an ETF domain boundary that likely affects the dynamic properties of ETF in the crystal and in solution. We show that Arg-alpha237 is required for TMADH to structurally imprint the as-purified semiquinone form of wild-type ETF and that the ability of TMADH to facilitate this structural reorganization is lost following (i) redox cycling of ETF, or simple conversion to the oxidized form, and (ii) mutagenesis of Arg alpha237. We discuss this result in light of recent apparent conflict in the literature relating to the structural imprinting of wild-type ETF. Our studies support a mechanism of electron transfer by conformational sampling as advanced from our previous analysis of the crystal structure of the TMADH-2ETF complex [Leys, D. , Basran, J. , Sutcliffe, M. J., and Scrutton, N. S. (2003) Nature Struct. Biol. 10, 219-225] and point to a key role for the Tyr-442 (TMADH) and Arg-alpha237 (ETF) residue pair in transiently stabilizing productive electron transfer configurations. Our work also points to the importance of Arg-alpha237 in controlling the thermodynamics of electron transfer, the dynamics of ETF, and the protection of reducing equivalents following disassembly of the TMADH-2ETF complex. PMID- 18407659 TI - Interaction with amyloid beta peptide compromises the lipid binding function of apolipoprotein E. AB - Apolipoprotein (apo) E is an exchangeable apolipoprotein that plays an integral role in cholesterol transport in the plasma and the brain. It is also associated with protein misfolding or amyloid proteopathy of the beta amyloid peptide (Abeta) in Alzheimer's disease (AD) and cerebral amyloid angiopathy. The C terminal domain (CT) of apoE encompasses two types of amphipathic alpha helices: a class A helix (residues 216-266) and a class G* helix (residues 273-299). This domain also harbors high-affinity lipoprotein binding and apoE self-association sites that possibly overlap. The objective of this study is to examine if the neurotoxic oligomeric Abeta interacts with apoE CT and if this association affects the lipoprotein binding function of recombinant human apoE CT. Site specific fluorescence labeling of single cysteine-containing apoE CT variants with donor probes were employed to identify the binding of Abeta bearing an acceptor probe by intermolecular fluorescence resonance energy-transfer analysis. A higher efficiency of energy transfer was noted with probes located in the class A helix than with those located in the class G* helix of apoE CT. In addition, incubation of apoE CT with Abeta severely impaired the lipid binding ability and the overall amount of lipid-associated apoE CT. However, when apoE CT is present in a lipid-bound state, Abeta appears to be localized within the lipid milieu of the lipoprotein particle and not associated with any specific segments of the protein. When our data are taken together, they suggest that Abeta association compromises the fundamental lipoprotein binding function of apoE, which may have implications not only in terms of amyloid buildup but also in terms of the accumulation of cholesterol at extracellular sites. PMID- 18407660 TI - Low-frequency dynamics of Caldariomyces fumago chloroperoxidase probed by femtosecond coherence spectroscopy. AB - Ultrafast laser spectroscopy techniques are used to measure the low-frequency vibrational coherence spectra and nitric oxide rebinding kinetics of Caldariomyces fumago chloroperoxidase (CPO). Comparisons of the CPO coherence spectra with those of other heme species are made to gauge the protein-specific nature of the low-frequency spectra. The coherence spectrum of native CPO is dominated by a mode that appears near 32-33 cm(-1) at all excitation wavelengths, with a phase that is consistent with a ground-state Raman-excited vibrational wavepacket. On the basis of a normal coordinate structural decomposition (NSD) analysis, we assign this feature to the thiolate-bound heme doming mode. Spectral resolution of the probe pulse ("detuned" detection) reveals a mode at 349 cm(-1), which has been previously assigned using Raman spectroscopy to the Fe-S stretching mode of native CPO. The ferrous species displays a larger degree of spectral inhomogeneity than the ferric species, as reflected by multiple shoulders in the optical absorption spectra. The inhomogeneities are revealed by changes in the coherence spectra at different excitation wavelengths. The appearance of a mode close to 220 cm(-1) in the coherence spectrum of reduced CPO excited at 440 nm suggests that a subpopulation of five coordinated histidine ligated hemes is present in the ferrous state at a physiologically relevant pH. A significant increase in the amplitude of the coherence signal is observed for the resonance with the 440 nm subpopulation. Kinetics measurements reveal that nitric oxide binding to ferric and ferrous CPO can be described as a single-exponential process, with rebinding time constants of 29.4 +/- 1 and 9.3 +/- 1 ps, respectively. This is very similar to results previously reported for nitric oxide binding to horseradish peroxidase. PMID- 18407661 TI - EPR and ENDOR studies of cryoreduced compounds II of peroxidases and myoglobin. Proton-coupled electron transfer and protonation status of ferryl hemes. AB - The nature of the [Fe(IV)-O] center in hemoprotein Compounds II has recently received considerable attention, as several experimental and theoretical investigations have suggested that this group is not necessarily the traditionally assumed ferryl ion, [Fe(IV)=O]2+, but can be the protonated ferryl, [Fe(IV)-OH]3+. We show here that cryoreduction of the EPR-silent Compound II by gamma-irradiation at 77 K produces Fe(III) species retaining the structure of the precursor [Fe(IV)=O]2+ or [Fe(IV)-OH]3+, and that the properties of the cryogenerated species provide a report on structural features and the protonation state of the parent Compound II when studied by EPR and 1H and 14N ENDOR spectroscopies. To give the broadest view of the properties of Compounds II we have carried out such measurements on cryoreduced Compounds II of HRP, Mb, DHP and CPO and on CCP Compound ES. EPR and ENDOR spectra of cryoreduced HRP II, CPO II and CCP ES are characteristic of low-spin hydroxy-Fe(III) heme species. In contrast, cryoreduced "globins", Mb II, Hb II, and DHP II, show EPR spectra having lower rhombicity. In addition the cryogenerated ferric "globin" species display strongly coupled exchangeable (1)H ENDOR signals, with A max approximately 20 MHz and a iso approximately 14 MHz, both substantially greater than for hydroxide/water ligand protons. Upon annealing at T > 180 K the cryoreduced globin compounds II relax to the low-spin hydroxy-ferric form with a solvent kinetic isotope effect, KIE > 6. The results presented here together with published resonance Raman and Mossbauer data suggest that the high-valent iron center of globin and HRP compounds II, as well as of CCP ES, is [Fe(IV)=O]2+, and that its cryoreduction produces [Fe(III)-O]+. Instead, as proposed by Green and co-workers, CPO II contains [Fe(IV)-OH]3+ which forms [Fe(III)-OH]2+ upon radiolysis. The [Fe(III)-O]+ generated by cryoreduction of HRP II and CCP ES protonate at 77 K, presumably because the heme is linked to a distal-pocket hydrogen bonding/proton-delivery network through an H-bond to the "oxide" ligand. The data also indicate that Mb and HRP compounds II exist as two major conformational substates. PMID- 18407662 TI - Anopheles gambiae cadherin AgCad1 binds the Cry4Ba toxin of Bacillus thuringiensis israelensis and a fragment of AgCad1 synergizes toxicity. AB - A midgut cadherin AgCad1 cDNA was cloned from Anopheles gambiae larvae and analyzed for its possible role as a receptor for the Cry4Ba toxin of Bacillus thuringiensis strain israelensis. The AgCad1 cadherin encodes a putative 1735 residue protein organized into an extracellular region of 11 cadherin repeats (CR) and a membrane-proximal extracellular domain (MPED). AgCad1 mRNA was detected in midgut of larvae by polymerase chain reaction (PCR). The AgCad1 protein was localized, by immunochemistry of sectioned larvae, predominately to the microvilli in posterior midgut. The localization of Cry4Ba binding was determined by the same technique, and toxin bound microvilli in posterior midgut. The AgCad1 protein was present in brush border membrane fractions prepared from larvae, and Cry4Ba toxin bound the same-sized protein on blots of those fractions. The AgCad1 protein was expressed transiently in Drosophila melanogaster Schneider 2 (S2) cells. 125I-Cry4Ba toxin bound AgCad1 from S2 cells in a competitive manner. Cry4Ba bound to beads extracted 200 kDa AgCad1 and a 29 kDa fragment of AgCad1 from S2 cells. A peptide containing the AgCad1 region proximal to the cell (CR11-MPED) was expressed in Escherichia coli. Although Cry4Ba showed limited binding to CR11-MPED, the peptide synergized the toxicity of Cry4Ba to larvae. AgCad1 in the larval brush border is a binding protein for Cry4Ba toxin. On the basis of binding results and CR11-MPED synergism of Cry4Ba toxicity, AgCad1 is probably a Cry4Ba receptor. PMID- 18407663 TI - Membrane insertion pathway of annexin B12: thermodynamic and kinetic characterization by fluorescence correlation spectroscopy and fluorescence quenching. AB - Experimental determination of the free energy stabilizing the structure of membrane proteins in their native lipid environment is undermined by the lack of appropriate methods and suitable model systems. Annexin B12 (ANX) is a soluble protein which reversibly inserts into lipid membranes under mildly acidic conditions, which makes it a good experimental model for thermodynamic studies of folding and stability of membrane proteins. Here we apply fluorescence correlation spectroscopy for quantitative analysis of ANX partitioning into large unilamellar vesicles containing either 25% or 75% anionic lipids. Membrane binding of ANX results in changes of autocorrelation time and amplitude, both of which are used in quantitative analysis. The thermodynamic scheme describing acid induced membrane interactions of ANX considers two independent processes: pH dependent formation of a membrane-competent form near the membrane interface and its insertion into the lipid bilayer. Our novel fluorescence lifetime topology method demonstrates that the insertion proceeds via an interfacial refolded intermediate state, which can be stabilized by anionic lipids. Lipid titration measurements are used to determine the free energy of both transmembrane insertion and interfacial penetration, which are found to be similar, approximately -10-12 kcal/mol. The formation of the membrane-competent form, examined in a lipid saturation experiment, was found to depend on the local proton concentration near the membrane interface, occurring with pK = 4.3 and involving the protonation of two residues. Our results demonstrate that fluorescence correlation spectroscopy is a convenient tool for the quantitative characterization of the energetics of transmembrane insertion and that pH triggered ANX insertion is a useful model for studying the thermodynamic stability of membrane proteins. PMID- 18407664 TI - Structural changes of horse heart ferricytochrome C induced by changes of ionic strength and anion binding. AB - To test the validity of the notion that changes in ionic strength and ion binding do not cause any major functionally relevant structural changes in cytochrome c, we measured the absorption and electronic circular dichroism (ECD) of horse heart ferricytochrome c for the Soret and 695 nm charge-transfer band as a function of dihydrogen phosphate and sodium acetate concentrations. This band is known to probe the integrity of the functionally pivotal Fe3+-M80 linkage. Spectral changes indicate that an ionic strength increase (via an increasing acetate ion concentration) affects only a subset of conformational substates of the Fe-M80 interface, probed by the 695 nm charge-transfer band, without a substantial modification of the heme environment. This result suggests that the substates probed by the 695 nm band differ with respect to their capability to transduce changes of solvent-protein interactions to the active site. The binding of H2PO4- ions causes more significant structural changes, which give rise to a large increase of the oscillator strength of the 695 nm band. This reflects a strengthening of the Fe-M80 bond in all substates, which probably destabilizes the oxidized state but stabilizes the folded state of the protein. Additional structural variations are likely to involve aromatic side chains, such as F82 and W59, and the hydrogen-bonding network in the heme pocket. In contrast to the current belief that anion binding to the binding domain of the protein for cytochrome c oxidase does not cause any functionally relevant structural changes, our results show that the structural variations that occur in the heme pocket are most likely of functional significance. PMID- 18407665 TI - Structure and stability changes of human IgG1 Fc as a consequence of methionine oxidation. AB - The Fc region has two highly conserved methionine residues, Met 33 (C(H)3 domain) and Met 209 (C(H)3 domain), which are important for the Fc's structure and biological function. To understand the effect of methionine oxidation on the structure and stability of the human IgG1 Fc expressed in Escherichia coli, we have characterized the fully oxidized Fc using biophysical (DSC, CD, and NMR) and bioanalytical (SEC and RP-HPLC-MS) methods. Methionine oxidation resulted in a detectable secondary and tertiary structural alteration measured by circular dichroism. This is further supported by the NMR data. The HSQC spectral changes indicate the structures of both C(H)2 and C(H)3 domains are affected by methionine oxidation. The melting temperature (Tm) of the C(H)2 domain of the human IgG1 Fc was significantly reduced upon methionine oxidation, while the melting temperature of the C(H)3 domain was only affected slightly. The change in the C(H)2 domain T m depended on the extent of oxidation of both Met 33 and Met 209. This was confirmed by DSC analysis of methionine-oxidized samples of two site specific methionine mutants. When incubated at 45 degrees C, the oxidized Fc exhibited an increased aggregation rate. In addition, the oxidized Fc displayed an increased deamidation (at pH 7.4) rate at the Asn 67 and Asn 96 sites, both located on the C(H)2 domain, while the deamidation rates of the other residues were not affected. The methionine oxidation resulted in changes in the structure and stability of the Fc, which are primarily localized to the C(H)2 domain. These changes can impact the Fc's physical and covalent stability and potentially its biological functions; therefore, it is critical to monitor and control methionine oxidation during manufacturing and storage of protein therapeutics. PMID- 18407666 TI - Mutational analysis of putative helix-helix interacting GxxxG-motifs and tryptophan residues in the two-peptide bacteriocin lactococcin G. AB - The membrane-permeabilizing two-peptide bacteriocin lactococcin G consists of two different peptides, LcnG-alpha and LcnG-beta. The bacteriocin contains several tryptophan and tyrosine residues and three putative helix-helix interacting GxxxG motifs, G 7xxxG 11 and G 18xxxG 22 in LcnG-alpha and G 18xxxG 22 in LcnG-beta. The tryptophan and tyrosine residues and residues in the GxxxG-motifs were altered by site-directed mutagenesis to analyze the structure and membrane orientation of lactococcin G. Substituting the glycine residues at position 7 or 11 in the G 7xxxG 11-motif in LcnG-alpha with large hydrophobic or hydrophilic residues was highly detrimental, whereas small residues were tolerated. Qualitatively similar results were obtained for the G 18xxxG 22-motif in LcnG beta. In contrast, replacement of the glycine residues in the middle of these two motifs with large hydrophilic residues was tolerated. All mutations in the G 18xxxG 22-motif in LcnG-alpha were relatively well-tolerated, indicating that this motif is not involved in helix-helix interactions. The four aromatic residues in the N-terminal part of LcnG-beta could individually be replaced by other aromatic residues, a hydrophilic positive residue, and a hydrophobic residue without a marked reduced activity, indicating that this region is structurally flexible and not embedded in a strictly hydrophobic or hydrophilic environment. The results are in accordance with a structural model where the G 7xxxG 11-motif in LcnG-alpha and the G 18xxxG 22-motif in LcnG-beta interact and allow the two peptides to form a parallel transmembrane helix-helix structure, with the tryptophan-rich N-terminal part of LcnG-beta positioned in the outer membrane interface and the cationic C-terminal end of LcnG-alpha inside the cell. PMID- 18407667 TI - Interaction of surfactant protein A with the intermediate filaments desmin and vimentin. AB - Surfactant protein A (SP-A), a member of the collectin family that modulates innate immunity, has recently been involved in the physiology of reproduction. Consistent with the activation of ERK-1/2 and COX-2 induced by SP-A in myometrial cells, we reported previously the presence of two major proteins recognized by SP A in these cells. Here we identify by mass spectrometry one of these SP-A targets as the intermediate filament (IF) desmin. In myometrial preparations derived from desmin-deficient mice, the absence of binding of SP-A to any 50 kDa protein confirmed the identity of this SP-A-binding site as desmin. Our data based on partial chymotrypsin digestion of pure desmin suggested that SP-A recognizes especially its rod domain, which is known to play an important role during the assembly of desmin into filaments. In line with that, electron microscopy experiments showed that SP-A inhibits in vitro the polymerization of desmin filaments. SP-A also recognized in vitro polymerized filaments in a calcium dependent manner at a physiological ionic strength but not the C1q receptor gC1qR. Furthermore, Texas Red-labeled SP-A colocalized with desmin filaments in myometrial cells. Interestingly, vimentin, the IF characteristic of leukocytes, is one of the major proteins recognized by SP-A in protein extracts of U937 cells after PMA-induced differentiation of this monocytic cell line. Interaction of SP A with vimentin was further confirmed using recombinant vimentin in solid-phase binding assays. The ability of SP-A to interact with desmin and vimentin, and to prevent polymerization of desmin monomers, shed light on unexpected and wider biological roles of this collectin. PMID- 18407668 TI - Isoelectric focusing in a microfluidically defined electrophoresis channel. AB - A new form of microchip isoelectric focusing that allows efficient coupling with pretreatment processes is reported. The sample is conveyed in a carrier ampholyte solution to the separation channel that is connected at both ends by two V-shaped lead channels, which supply electrode solutions to the connection point and complete the electrical connection to off-chip electrodes. The relatively high electric conductivity of the electrode solutions compared with that of the pH gradient enables focusing with a 2% loss of applied voltage at the electrodes using the lead channels. A glass microchip was constructed specifically for this configuration. The channel wall was coated with polydimethylacrylamide, and the IEF chip was operated in a chip holder equipped with on-chip connector valves. A plug of fluorescence-labeled peptide p I markers with p I values ranging from 3.64 to 9.56 with carrier ampholyte solution (pH 3-10) was introduced into the separation channel. When the plug reached the channel segment (24 mm in length) between the connection points with the electrolyte lead channels, isoelectric focusing was started after filling the lead channels with electrolyte solutions. The peptide markers were observed using scanning fluorescence detection. The entire range of the pH gradient was established in the segment after approximately 2 min. Isoelectric focusing of three consecutively injected sample plugs containing different p I markers was demonstrated. PMID- 18407669 TI - Speedy component resolution: an improved tool for processing diffusion-ordered spectroscopy data. AB - Diffusion-based NMR techniques (e.g., diffusion-ordered spectroscopy, DOSY), which can be used to distinguish between the signals of different components of a mixture, are steadily gaining in popularity. When processing data from a DOSY experiment it is often desirable to reconstruct the spectra of individual components; here, multivariate methods that take advantage of the covariance between the resonances of a given component can often be advantageous. This paper presents a minor variation on the established CORE method, speedy component resolution (SCORE), that gives a major improvement in performance. In common with CORE it can use any experimental sampling scheme and is adaptable to different experimental decay shapes, but unlike CORE it is very fast and relatively insensitive to starting guesses. The method is demonstrated on a mixture of quinine, geraniol, and camphene in deuteriated methanol, where all four component spectra can be extracted in less than 15 s. PMID- 18407670 TI - Exploring the mechanism of selective noncovalent adduct protein probing mass spectrometry utilizing site-directed mutagenesis to examine ubiquitin. AB - Mass spectrometry (MS) is emerging as an additional tool for examining protein structure by way of experiments where structurally related mass changes induced in solution are subsequently detected in the gas phase. Selective noncovalent adduct protein probing (SNAPP) is a recent addition to this type of experiment. SNAPP utilizes noncovalent recognition of lysine residues with 18-crown-6 (18C6) to monitor changes in protein structure. It has been observed that the number of 18C6 adducts that attach to a protein is a function of the structure of the protein. The present work seeks to examine the underlying chemistry which controls the differential attachment of 18C6 to lysine by using ubiquitin as a model system. Ubiquitin is a small protein with a structure that has been well characterized by multiple techniques. Site-directed mutagenesis was used to create a series of ubiquitin mutants where the lysine residues were exchanged for asparagine one at a time. These mutants were then evaluated by SNAPP-MS to determine the relative contribution of each lysine as a binding site for 18C6. It was found that attachment of 18C6 is largely controlled by the strength of intramolecular interactions involving lysine residues. Salt bridges provide the greatest interference, followed by hydrogen bonds. In addition to determining the mechanism for SNAPP, insights are provided about the structure of ubiquitin including confirmation of the existence of two dynamic states for the native structure. These results are discussed in relation to the biological functions of ubiquitin. PMID- 18407671 TI - Bioassembled nanocircuits of Mo6S9-xIx nanowires for electrochemical immunodetection of estrone Hapten. AB - We demonstrate a novel and highly sensitive electrochemical detection of estrone based on an immunosensor platform, composed of bioassembled nanocircuits of Mo 6S 9- x I x nanowires (MoSI NWs) covalently connected to anti-estrone antibodies. The one-step, label-free, and quantitative detection of estrone is realized by employing the [Ru(NH 3) 6] (3+/2+) redox ions to sense anti-estrone antibody and estrone interactions. The MoSI NWs/anti-estrone antibody nanocircuit architectures provide an amplification and conductive pathway for the specific electrochemical sensing of estrone hapten. A detection limit of 1.4 pg x mL (-1) was achieved in contrast to previous electrochemical techniques in which the sensitivity was limited to the nanomolar range. PMID- 18407672 TI - Microfluidic analogy of the wheatstone bridge for systematic investigations of electro-osmotic flows. AB - A microfluidic analogy of the electric Wheatstone Bridge has been developed for electrokinetic study of miscellaneous liquid-solid interfaces. By using an optimized glass-PDMS-glass device technology, microfluidic channels with well controlled surface properties can be fabricated, forming an "H" shaped fluidic network. After solving a set of linear equations, the electro-osmotic flow rate in the center channel can be deduced from indirect measurement of flow rates in the lateral channels. Experimentally, we demonstrate that the electro-osmotic mobility can be monitored every 30 s with accuracy better than 3% for a large dynamic range of electric fields. The results obtained with a borosilicate glass (D-263) and several standard biological buffers are also shown to illustrate the capability of this high throughput method. PMID- 18407673 TI - Exploring Mn-doped ZnS quantum dots for the room-temperature phosphorescence detection of enoxacin in biological fluids. AB - While most research works focus on the development of quantum dots (QDs)-based fluorescence sensors, much less attention is paid to the phosphorescence properties of QDs and their potential for phosphorescence detection. In this work, the phosphorescence property of Mn-doped ZnS QDs is explored to develop a novel room-temperature phosphorescence (RTP) method for the facile, rapid, cost effective, sensitive, and selective detection of enoxacin in biological fluids. The Mn-doped ZnS QDs-based RTP method reported here does not need the use of deoxidants and other inducers and allows the detection of enoxacin in biological fluids without interference from autofluorescence and the scattering light of the matrix. The Mn-doped ZnS QDs offer excellent selectivity for detecting enoxacin in the presence of the main relevant metal ions in biological fluids, biomolecules, and other kinds of antibiotics. Quenching of the phosphorescence emission due to the addition of enoxacin at 1.0 microM is unaffected by 5000-fold excesses of Na (+) and 10000-fold excesses of K (+), Mg (2+), and Ca (2+). Amino acids such as tryptophan, histidine, and l-cysteine at 1000-fold concentration of enoxacin do not affect the detection of enoxacin. Glucose does not affect the detection at 10000-fold concentration of enoxacin. Typical coadministers (mainly other types of antibiotics) such as ceftezole, cefoperazone, oxacillin, and kalii dehydrographolidi succinas are permitted at 50-, 10-, 100-, and 50-fold excesses, respectively, without interference with the detection of enoxacin. The precision for 11 replicate detections of 0.4 microM enoxacin is 1.8% (RSD). The detection limit for enoxacin is 58.6 nM. The recovery of spiked enoxacin in human urine and serum samples ranges from 94 to 104%. The developed Mn-doped ZnS QDs-based RTP method is employed to monitor the time-dependent concentration of enoxacin in urine from a healthy volunteer after the oral medication of enoxacin. The investigation provides evidence that doped QDs are promising for RTP detection in further applications. PMID- 18407674 TI - Femtomolar electrochemical detection of DNA hybridization using hollow polyelectrolyte shells bearing silver nanoparticles. AB - The preparation, and use as electrochemical labels, of polyelectrolyte shells bearing Ag nanoparticles is described. Their potential for highly sensitive detection is demonstrated. The shells are prepared by layer-by-layer self assembly around templates (500 nm diameter) which are then dissolved. The shells can be opened and closed by adjustment of solution pH, and this process is utilized to encapsulate Ag nanoparticles, chiefly by adsorption to the inner walls of the capsules. Based on absorbance, TEM and voltammetric measurements, the highest loading achieved is approximately 78 Ag particles per capsule. The Ag capsules are used via biotin-avidin binding as labels for the detection of DNA hybridization, following acid dissolution and then detection of the Ag (+) by ASV. A 30-mer sequence specific to Escherichia coli is measured at DNA-modified screen-printed electrodes with a detection limit of approximately 25 fM, which corresponds to the detection of 4.6 fg ( approximately 3 x 10 (5) molecules) in the 20 microL analyte sample. A 200 fM target containing a single mismatch gives a significantly (<74%) lower response than 200 fM of complementary target; 60 pM of noncomplementary target gives a negligible response. PMID- 18407675 TI - Assessment of the metabolism and intrinsic reactivity of a novel catechol metabolite. AB - PH-302 ( 1) demonstrates potent inhibitory activity against the inducible form of nitric oxide synthase (iNOS). The primary metabolite of PH-302 is a catechol ( 2) resulting from oxidative demethylenation of the methylenedioxyphenyl moiety by cytochrome P450 3A4. Concerns regarding subsequent two-electron oxidation of 2 to an electrophilic quinone species and the potential for resulting toxicity prompted additional studies to examine the reactivity and metabolic fate of this metabolite. Contrary to literature reports of catechol reactivity, 2 appeared to be resistant to quinone formation in human liver microsomal incubations, as determined by the lack of detectable glutathione (GSH) adducts and no covalent binding to microsomal proteins. In addition, 2 showed no evidence of depletion of intracellular glutathione or cytotoxicity at concentrations up to 1 mM in primary human and rat hepatocytes. In the presence of tyrosinase, spectral evidence indicated that 2 was oxidized to the ortho-quinone, and upon incubation in the presence of GSH, two conjugates were detected and characterized by LC/MS/MS. Lastly, the metabolic pathways of 2 were investigated in rat and human hepatocytes and found to be similar, proceeding via glucuronidation, sulfation, and methylation of the catechol. Collectively, these studies demonstrate that 2 appears to be resistant to further oxidation to quinone in liver microsomes, as well as spontaneous redox cycling, while the formation of phase II metabolites in hepatocytes suggests that multiple detoxication pathways may provide added protection against toxicity in the liver. PMID- 18407676 TI - Biochemical assays of immobilized oligonucleotides with mass spectrometry. AB - This paper reports the use of mass spectrometry to characterize oligonucleotides immobilized to the surfaces of biochips. Biotinylated oligonucleotides were immobilized to self-assembled monolayers that present a streptavidin layer and then treated with a complementary strand to present short duplexes. Treatment of the surface with 5-methoxysalicylic acid and ammonium citrate matrix allows the individual oligonucleotides to be observed by matrix-assisted laser desorption/iozation and time-of-flight mass spectrometry (MALDI-TOF MS). Examples are shown wherein this method is applied to assays of hybridization, of cleavage by a deoxyribozyme, of a dephosphorylation reaction, and of the adducts formed on treatment of DNA with cis-platin. This work provides an early example of the application of mass spectrometry to DNA biochips and may substantially expand the applications of the now common oligonucleotide arrays. PMID- 18407677 TI - Copolymer-induced stabilizing effect of highly swollen hexagonal mesophases. AB - We show that small amounts of copolymer that decorate an oil/water interface can greatly enhance the stability of swollen surfactant hexagonal phases, comprising oil tubes regularly arranged in a water matrix. Both the radius of the tubes and the thickness of the aqueous channel between the tubes can be controlled independently over large ranges. Such soft composite materials offer a potential interest for the synthesis of mesoporous materials. PMID- 18407678 TI - Dewetting of polystyrene thin films on poly(ethylene glycol)-modified surfaces as a simple approach for patterning proteins. AB - A simple technique for patterning proteins utilizing dewetted polystyrene (PS) droplets is demonstrated. A polystyrene thin film was spin coated on a poly(ethylene glycol) (PEG) silane-modified surface. As the PS film dewets from the surface, upon annealing, to form droplets, the PEG-silane-modified surface is exposed, which retains its capability to resist protein adsorption, and the PS droplets allow the selective adsorption of proteins. In contrast to the undewetted flat PS film, the droplet surface had a greater amount of adsorbed proteins. Atomic force microscopy scans reveal that the roughness of the droplet surface is higher, and a multilayer of proteins results on the droplet surface. Moreover, micro- and nanoscale droplet patterns can easily be achieved by tuning the thickness of PS thin films. Because dewetting approaches for generating ordered dewetting droplets have been successfully generated by others, those approaches could be easily combined with this technique to fabricate ordered protein patterns. PMID- 18407679 TI - Surface-templated formation of protein microfibril arrays. AB - Ordered arrays of collagen microfibrils form rapidly and spontaneously from a solution of monomers deposited onto a mica substrate. These arrays are well ordered and apparently continuous over the entire substrate. Correlated atomic force microscope images and Laue diffraction patterns indicate that the protein alignment and microfibril formation is controlled by the crystal orientation of the mica substrate rather than fluid flow or drying effects. This surface-induced mechanism allows for immediate, robust, and reproducible pattern formation. PMID- 18407680 TI - Stimuli-responsive supramolecular nanostructure from amphiphilic calix[4]arene and its three-dimensional dendritic silver nanostructure. AB - We synthesized a tetrameric amphiphilic molecule ( 1) based on a calix[4]arene building block that self-assembled into different tunable and stable aggregation structures at different pH values in aqueous solution. The amphiphilic calix[4]arene molecule ( 1) formed a spherical structure at pH 3. However, 1 formed a hollow necklacelike structure of 500 nm diameter at pH 7. These results indicate that the self-assembled morphologies of 1 are strongly dependent on pH values. In addition, a 3D dendritic silver nanostructure was obtained by the self assembly of 1 at pH 7. PMID- 18407681 TI - Synthesis and screening of a library of Re/Tc-based amyloid probes derived from beta-breaker peptides. AB - Through the development and application of a unique approach for producing Re metallopeptides, a new class of peptide-derived probes that are designed to target beta-amyloid plaques was developed. Derivatives of a class of beta-breaker peptides having the core sequence lvffa or affvl (lower case letters represent D amino acids) and the single amino acid chelate quinoline (SAACQ) ligand which can bind Re and (99m)Tc were prepared on an automated peptide synthesizer. Both monomeric and dimeric peptides were synthesized in modest to good yields where in select examples a biotin-containing amino acid derivative was included to act as a linker point for further conjugation to carrier proteins. The Re complexes for all reported peptides were prepared similarly and screened for their ability to inhibit fibrillogenesis. Two of the reported compounds showed excellent inhibitory properties (8a: 40 +/- 5% amyloid formation versus control; 16: 40 +/- 4%) and warrant further investigation. For one of these leads, the (99m)Tc analogue was synthesized and the product showed high stability toward histidine and cysteine challenges, making it a viable candidate for in vivo biodistribution studies. PMID- 18407682 TI - Synthesis and proteomic activity evaluation of a new isotope-coded affinity tagging (ICAT) reagent. AB - During recent years, quantitative proteome profiling has taken advantage of incorporating the traditional stable isotope dilution analysis into global scale or discovery-based proteomic experiments that use mass spectrometers as detectors to allow the pairwise study of differently expressed proteins. Quantitative protein analysis by means of the isotope-coded affinity tag (ICAT) method and tandem mass spectrometry (MS) enables the pairwise comparison of protein expression levels in biological samples. Herein, a modified ICAT reagent, named BAA-ICAT (beta-alanine-arm-ICAT) in which the polyether linker is replaced by a more water-soluble polyamide one, was investigated. PMID- 18407683 TI - Direct site-specific labeling of the Cys-tag moiety in scVEGF with technetium 99m. AB - Angiogenesis is a fundamental feature of tumor development, and therefore, the tracers for molecular imaging of specific angiogenic biomarkers are expected to be useful for diagnostics, patient monitoring, and drug development. We have created a new class of imaging agents based on the most important mediator of angiogenesis, vascular endothelial growth factor (VEGF). Our latest version is a single-chain (sc) VEGF protein containing an N-terminal Cys-tag designed for site specific modification with a variety of imaging and therapeutic moieties. We have recently found that the Cys-tag itself can form a stable chelate with (99m)Tc using tin-tricine as an exchange reagent. This self-chelation approach yields a highly stable and fully functional form of radiolabeled scVEGF that can be used as a SPECT tracer for tumor angiogenesis. Also of note is that directly labeled scVEGF has less than one-half the nonspecific renal uptake of (99m)Tc-HYNIC scVEGF. The simple production of scVEGF for direct chelation of (99m)Tc makes it a promising molecular imaging agent for the oncology clinic. PMID- 18407684 TI - Prognostic significance of head-and-neck cancer biomarkers previously discovered and identified using iTRAQ-labeling and multidimensional liquid chromatography tandem mass spectrometry. AB - Diagnostic oncoproteomics is an emerging field; at present, studies on evaluation of prognostic utility of potential biomarkers identified using proteomic techniques are limited. Analysis with isobaric mass tags (iTRAQ) by multidimensional liquid chromatography-mass spectrometry (LC-MS/MS) to identify proteins that are differentially expressed in human head-and-neck/oral squamous cell carcinomas (HNOSCCs) versus noncancerous head-and-neck tissues has led to the discovery, identification, and verification of consistently increased expression of a panel of proteins, including stratifin (14-3-3sigma) and YWHAZ (14-3-3zeta), that may serve as potential cancer biomarkers. Herein, we describe the prognostic utility of these two candidate biomarkers for head-and-neck/oral squamous cell carcinoma (HNOSCC). To determine the clinical significance of stratifin and YWHAZ in head-and-neck tumorigenesis, the expressions of these two proteins were analyzed in HNOSCCs (51 cases) and nonmalignant tissues (39 cases) using immunohistochemistry. Significant increase in stratifin expression was observed in the HNOSCCs as compared to the nonmalignant mucosa [p=0.003, Odd's Ratio (OR)=3.8, 95% CI=1.6-9.2]. Kaplan-Meier survival analysis reveals correlation of stratifin overexpression with reduced disease-free survival of HNOSCC patients (p=0.06). The most intriguing finding is the significant decrease in median disease-free survival (13 months) in HNOSCC patients showing overexpression of both stratifin and YWHAZ proteins, as compared to patients that did not show overexpression of these proteins (median disease-free survival=38 months, p=0.019), underscoring their utility as adverse prognosticators for HNOSCCs. Co-immunoprecipitation assays show the formation of stratifin-YWHAZ heterodimers in HNOSCC cells and tissue samples, and interactions with NFkappaB, beta-catenin, and Bcl-2 proteins. These results suggest the involvement of these proteins in the development of head-and-neck cancer and their association with adverse disease outcome. PMID- 18407685 TI - SALAMI (Spectrum ALignments using high Accuracy Mass and hIgh sensitivity data): how to make the best out of hybrid MS/MS data. AB - We present a software algorithm that combines ion trap and orbitrap product ion spectra acquired in parallel. The hybrid product ion spectra identify more peptides than when using two separate searches for the orbitrap and LTQ data. The program extracts the high-accuracy mass data from the Orbitrap mass analyzer and combines it with the high-sensitivity data analyzed in the LTQ linear ion trap; the m/z values of the high-confidence fragment ions are corrected to orbitrap mass accuracies and the fragment ion intensities are amplified. This approach utilizes the parallel spectrum measurement capabilities of the LTQ-Orbitrap. We present our approach to handling this type of hybrid data, explain our alignment program, and discuss the advantages of the chosen methodology. PMID- 18407686 TI - Characterization of the epithelial permeation enhancing effect of basic butylated methacrylate copolymer--in vitro studies. AB - Membrane destabilizing properties and increased efflux of doxorubicin from liposomes caused by basic butylated methacrylate copolymer (BBMC), better known under its commercial trade name EUDRAGIT E, have been described in the scientific literature. Here, we investigated the effect of BBMC on suspended and filter grown Caco2 cells with respect to apical-to-basal transport and membrane permeabilization using transport assays, trypan blue exclusion assay, measurements of transepithelial electrical resistance (TEER), confocal laser scanning microscopy, and transmission electron microscopy. The effect of inhibiting protein phosphatase 2A (PP2A) by okadaic acid was investigated by measuring TEER, but a link between PP2A and the observed effects could not be established. Overall, membrane permeabilization of Caco2 cells by BBMC was demonstrated, which went along with increased apical-to-basal transport of the model compounds mannitol, talinolol, and trospium. The effect was concentration- and time-dependent, and reversible. Enhancement occurred at polymer concentrations as low as 20 microg/mL. PMID- 18407687 TI - Copper-catalyzed coupling of tertiary aliphatic amines with terminal alkynes to propargylamines via C-H activation. AB - We have developed a convenient and efficient method for coupling of tertiary aliphatic amines with terminal alkynes to propargylamines via C-H activation. The protocol uses CuBr as the catalyst, NBS as the free radical initiator, CH(3)CN as the solvent, and the alkynylation was selectively performed on the methyl of tertiary aliphatic amines at 80 degrees C. This is an economical and practical method for the synthesis of propargylamines. PMID- 18407689 TI - N,N'-disubstituted methylenediimidazolium salts: a versatile guest for various macrocycles. AB - N,N'-Disubstituted methylenediimidazolium salts allow the formation of flexible inclusion complexes with beta-cyclodextrin, cucurbit[7]uril, tetrapropoxycalix[4]arene, and dibenzo-24-crown-8 ether. Due to the salt nature of the imidazolium guest, the counterion largely determines its solubility in a given solvent. Moreover, by the judicious choice of the imidazolium substituents, inclusion complexes of guest salts were obtained with a variety of macrocyclic hosts, and the binding parameters of the inclusion were determined for each complex. PMID- 18407690 TI - Cation-induced pi-stacking. AB - The design and synthesis of a new dipyridyl ligand with appended phenanthryl moieties is described. On addition of increments of silver(I) trifluoromethanesulfonate to a solution of the ligand, the phenanthryl protons shift upfield in the (1)H NMR spectrum, suggesting that the phenanthrenes pi stack on coordination of silver(I). In accord with this, the oxidation potential decreased from 1.74 to 1.55 V on complexation of silver(I). The pi-stacking was confirmed with the single-crystal X-ray structure of a palladium(II) coordination complex. PMID- 18407691 TI - Dihydroxystyrene metabolites from an association of the sponges Poecillastra wondoensis and Jaspis sp. AB - Five new dihydroxystyrene metabolites and six known compounds of the same structural class were isolated from an association of the sponges Poecillatra wondoensis and Jaspis sp., collected from Keomun Island, Korea. The structures of novel compounds were determined to be the sodium or N, N-dimethyl guanidinium salts of a dihydroxystyrene dimer (5) and two trimers (6, 7). Two dimers (10, 11) containing imidazole moieties were also identified on the basis of the results of combined spectroscopic analyses. Several compounds exhibited weak to moderate inhibitory effects against isocitrate lyase and sortase A enzymes derived from microorganisms. PMID- 18407692 TI - Spongian diterpenes with thyrotropin releasing hormone receptor 2 binding affinity from Spongia sp. AB - High-throughput screening of a plant and marine invertebrate extract library to find natural products with rat thyrotropin releasing hormorne (TRH) receptor 2 binding affinity led to the isolation of four new (1-4) and one known (5) spongian diterpene from the sponge Spongia sp. The structures were assigned from interpretation of 2D NMR and high-resolution ESIMS data. The absolute configurations of 1-4 were proposed on the basis of analysis of their CD spectra. Diterpenes 1-5 showed rat TRH receptor 2 binding affinity with IC(50) values of 23 microM, 70 microM, 400 microM, 600 microM, and 1 mM, respectively. PMID- 18407693 TI - Kahalalides V-Y isolated from a Hawaiian collection of the sacoglossan mollusk Elysia rufescens. AB - Four new kahalalides, V (1), W (2), X (3), and Y (4), as well as six previously characterized kahalalides have been isolated from a two-year collection of the sacoglossan mollusk Elysia rufescens. Curiously, kahalalide B, previously isolated in high yield from E. rufescens, was found to be essentially absent from these collections despite identical collection sites and times with previous collections. In addition, kahalalide K, which to date has only been reported from Bryopsis sp., was found in this collection of E. rufescens, suggesting that the production of these metabolites could potentially be from a microbial association with the mollusk and algae, and this relationship is continuously evolving in response to changes in the environment and predation. The structures of new peptides have been established on the basis of extensive 1D and 2D NMR spectroscopic data analysis. Kahalalide V (1) was ascertained to be an acyclic derivative of kahalalide D (5), while kahalalide W (2) was determined to have a 4 hydroxy-L-proline residue instead of the proline in 5. The arginine residue of kahalalide X (3), an acyclic derivative of kahalalide C, was determined to have an L configuration. Kahalalide Y (4) was found to have an L-proline residue instead of the hydroxyproline in kahalalide K. It is clear from this collection of E. rufescens that the discovery of new kahalalide-related metabolites is still highly feasible. PMID- 18407694 TI - Synthesis and cytotoxicity of a salicylihalamide A analogue. AB - The synthesis of a simple analogue of salicylihalamide A with a truncated lactone ring (+/- 2) was achieved in 10 steps. Its cytotoxicity profile in the NCI 60 cell-line human tumor assay differed significantly from that of salicylihalamide A in both level and specificity. PMID- 18407695 TI - Three-component combinatorial synthesis of novel dihydropyrano[2,3-c]pyrazoles. PMID- 18407696 TI - Parallel synthesis of 2-sulphanylated bis-benzimidazoles on soluble polymer support. AB - A well-sustained multistep synthetic protocol has been designed for the PEG functionalized aromatic acid amide to generate a molecular library of 2-alkylthio bis-benzimidazoles. An attempted synthesis of benzimidazole-2-thiol in dichloromethane has led to S-chloromethyl methyl sulfides, mimicking bacterial enzymatic systems. Regioselective S-alkylation was brought about under controlled conditions using a mild base at room temperature. The polymer-free compounds, 2 sulfanylated bisbenzimidazoles, were obtained in high yields and high purities. Chemical shift changes in proton and carbon NMR have been employed to monitor the progress of the reaction steps and to prove the site of S-alkylation, respectively. PMID- 18407697 TI - Randomized combinatorial library of heteroglycoclusters (hGC). PMID- 18407698 TI - Integration of ZnO microcrystals with tailored dimensions forming light emitting diodes and UV photovoltaic cells. AB - This article reports a new integration approach to produce arrays of ZnO microcrystals for optoelectronic and photovoltaic applications. Demonstrated applications are n-ZnO/p-GaN heterojunction LEDs and photovoltaic cells. The integration process uses an oxygen plasma treatment in combination with a photoresist pattern on magnesium doped GaN substrates to define a narrow sub-100 nm width nucleation region. Nucleation is followed by lateral epitaxial overgrowth producing single crystal disks of ZnO with desired size over 2 in. wafers. The process provides control over the dimensions (<1% STD) and the location (0.7% STD pitch variation) of the ZnO crystals. The quality of the patterned ZnO is high; the commonly observed defect related emission in the electroluminescence spectra is completely suppressed, and a single near-band-edge UV peak is observed. PMID- 18407699 TI - Large anisotropy of electrical properties in layer-structured In2Se3 nanowires. AB - Layer-structured indium selenide (In 2Se 3) nanowires (NWs) have large anisotropy in both shape and bonding. In 2Se 3 NWs show two types of growth directions: [11 20] along the layers and [0001] perpendicular to the layers. We have developed a powerful technique combining high-resolution transmission electron microscopy (HRTEM) investigation with single NW electrical transport measurement, which allows us to correlate directly the electrical properties and structure of the same individual NWs. The NW devices were made directly on a 50 nm thick SiN x membrane TEM window for electrical measurements and HRTEM study. NWs with the [11 20] growth direction exhibit metallic behavior while the NWs grown along the [0001] direction show n-type semiconductive behavior. Excitingly, the conductivity anisotropy reaches 10 (3)-10 (6) at room temperature, which is 1-3 orders magnitude higher than the bulk ratio. PMID- 18407700 TI - Photophysical properties of sulfonated dihydroxy phosphorus(V) tetrabenzotriazacorrole. AB - The photosensitizing properties of a novel phthalocyanine analogue, sulfonated dihydroxy phosphorus(V) tetrabenzotriazacorrole [P(OH)2TBCSn] and a non sulfonated one [P(OH)2TBC] are reported in this paper. Different from other phthalocyanine derivatives, P(OH)2TBCSn shows little aggregation in aqueous solution. The fluorescence quantum yield (PhiF) of P(OH)2TBCSn is lower than that of the non-sulfonated one. Studies of triplet state photophysics show that the presence of peripheral substituents on the macrocycle enhances the quantum yield of the triplet state. The sulfonated derivative, P(OH)2TBCSn, has a longer triplet lifetime (tauT = 0.234 ms) and higher singlet oxygen quantum yield (PhiDelta = 0.88) than P(OH)2TBC. Together with the ground-state absorption properties, the photosensitizing properties of the new compound suggest that it may be used as an excellent photosensitizer for photodynamic therapy (PDT). PMID- 18407701 TI - Calculation of vibrational transition frequencies and intensities in water dimer: comparison of different vibrational approaches. AB - We have calculated frequencies and intensities of fundamental and overtone vibrational transitions in water and water dimer with use of different vibrational methods. We have compared results obtained with correlation-corrected vibrational self-consistent-field theory and vibrational second-order perturbation theory both using normal modes and finally with a harmonically coupled anharmonic oscillator local mode model including OH-stretching and HOH bending local modes. The coupled cluster with singles, doubles, and perturbative triples ab initio method with augmented correlation-consistent triple-zeta Dunning and atomic natural orbital basis sets has been used to obtain the necessary potential energy and dipole moment surfaces. We identify the strengths and weaknesses of these different vibrational approaches and compare our results to the available experimental results. PMID- 18407702 TI - Molecular simulation of the effect of temperature and architecture on polyethylene barrier properties. AB - We present a multiscale approach for calculating the low-concentration solubility, diffusivity, and selectivity of small molecules through polymer matrixes. The proposed modeling scheme consists of two main stages; first, thoroughly equilibrated and representative poly(ethylene) (PE) atomistic melt configurations were obtained through the application of a Monte Carlo (MC) scheme based on advanced chain-connectivity altering moves (linear architectures) or the combination of localized MC moves followed by molecular dynamics. In the second phase, transition-state theory (TST), as proposed by Gusev and Suter [Gusev, A. A.; Suter, U. W. J. Chem. Phys. 1993, 99, 2228], was invoked in a coarser level of description to calculate the barrier properties of the studied macromolecules to small gas molecules at infinite dilution. The multiscale methodology was successfully applied on PE melts characterized by various molecular weights (MW) (from C78 up to C1000) and polydispersity indices at a wide range of temperature conditions. The effect of molecular architecture on the barrier properties was examined through the comparison between linear and short-chain branched structures bearing the same total number of carbon atoms. Simulation results were found to be in very good agreement with available experimental data. Additionally, the new scheme has been further validated by comparing the qualitative behavior of solubility, diffusivity, and selectivity with previously reported trends in the literature based on both experimental and simulation studies. The present study concludes that density plays a dominant role that determines the behavior of the polymer as a barrier material, especially in terms of diffusivity. Additionally, it is evidenced that short-chain branching has a small effect on the barrier properties of PE when the comparison is performed on purely amorphous samples. The hierarchical method presented here not only is faster when compared against conventional molecular dynamics simulations, but in some cases, like the vicinity of the glass transition temperature or for long polymer chain melts, it opens the way to the calculation of the barrier properties at realistic simulation times. PMID- 18407703 TI - Folded structures of L-leucylglycine oligopeptides and their aggregational behavior in aqueous solution: Raman scattering spectra and proton NMR spin lattice relaxation studies. AB - The aggregational behavior of three L-leucylglycine oligopeptides (residue numbers of glycine are 3, 4, and 5) in aqueous solution was investigated by the use of Raman scattering and 1H NMR spin-lattice relaxation methods. The results indicate that their oligopeptides take up a folded structure to form dimeric aggregates above their critical aggregation concentration. The application of one dimensional aggregate theory to these systems provides the following prediction. Elongation up to 6 glycine residues makes it possible to form dimeric aggregates, but further elongation (up to 7 glycine residues) makes the aggregates very unstable, and up to 8 or 9 glycine residues makes the formation of dimeric aggregates very difficult. The one-dimensional aggregate theory may be used to predict the existence of peptide aggregates through intermolecular hydrogen bonding. PMID- 18407704 TI - Mixed micelles of a PEO-PPO-PEO triblock copolymer (P123) and a nonionic surfactant (C12EO6) in water. a dynamic and static light scattering study. AB - The present article reports on static and dynamic light scattering (SLS and DLS) studies of aqueous solutions of the nonionic surfactant C12EO6 and the poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer EO20PO68EO20 (P123) at temperatures between 25 and 45 degrees C. In water, P123 self-assembles into spherical micelles with a hydrodynamic radius of 10 nm, and at 40 degrees C, these micelles consist of 131 unimers. Addition of C12EO6 leads to an association of the surfactant molecules to the P123 micelles and mixed micelles are formed. The size and structure of the mixed micelles as well as interparticle interactions were studied by varying the surfactant-to copolymer (C12EO6/P123) molar ratio. The novelty of this study consists of a composition-induced structural change of the mixed micelles at constant temperature. They gradually change from being spherical to polymer-like with increasing C12EO6 content. At low C12EO6/P123 molar ratios (below 12), the SLS measurements showed that the molar mass of the mixed micelles decreases with an increasing amount of C12EO6 in the micelles for all investigated temperatures. In this regime, the mixed micelles are spherical and the DLS measurements revealed a decrease in the hydrodynamic radius of the mixed micelles. An exception was found for C12EO6/P123 molar ratios between 2 and 3, where the mixed micelles become rodlike at 40 degrees C. This was the subject of a previous study and has hence not been investigated here. At high molar ratios (48 and above), the polymer-like micelles present a concentration-induced growth, similar to that observed in the pure C12EO6/water system. PMID- 18407705 TI - Comment on "The Kirkwood-Buff theory of solutions and the local composition of liquid mixtures". PMID- 18407707 TI - Homogeneous decomposition mechanisms of diethylzinc by Raman spectroscopy and quantum chemical calculations. AB - The gas-phase decomposition pathways of diethylzinc (DEZn), a common precursor for deposition of Zn-VI compounds, were investigated in detail. The homogeneous thermal decomposition of DEZn in N2 carrier was followed in an impinging-jet, up flow reactor by Raman scattering. Density Functional Theory calculations were performed to describe the bond dissociation behavior using the model chemistry B3LYP/6-311G(d) to estimate optimal geometries and Raman active vibrational frequencies of DEZn, as well as anticipated intermediates and products. Comparison of the measured DEZn decomposition profile to that predicted by a 2-D hydrodynamic simulation revealed that simple bond dissociation between zinc and carbon atoms is the dominant homogeneous thermal decomposition pathway. The calculations suggest several reactions involving intermediates and Raman scattering experiments confirming the formation of the dimer (ZnC2H5)2. In a different set of experiments, photolysis of DEZn gave evidence for decomposition by beta-hydride elimination. The results suggest that beta-hydride elimination is a minor pathway for the gas-phase homogeneous pyrolysis of diethylzinc. A reasonable transition state during beta-hydride elimination was identified, and the calculated energies and thermodynamic properties support the likelihood of these reaction steps. PMID- 18407708 TI - Molecular structure and dynamics in the low temperature (orthorhombic) phase of NH3BH3. AB - Variable temperature 2H NMR experiments on the orthorhombic phase of selectively deuterated NH3BH3 spanning the static to fast exchange limits of the borane and amine motions are reported. New values of the electric field gradient (EFG) tensor parameters have been obtained from the static 2H spectra of V(zz) = 1.652 (+/-0.082) x 10(21) V/m(2) and eta = 0.00 +/- 0.05 for the borane hydrogens and V(zz) = 2.883 (+/-0.144) x 10(21) V/m(2) and eta = 0.00 +/- 0.05 for the amine hydrogens. The molecular symmetry inferred from the observation of equal EFG tensors for the three borane hydrogens and likewise for the three amine hydrogens is in sharp contrast with the C(s) symmetry derived from diffraction studies. The origin of the apparent discrepancy has been investigated using molecular dynamics methods in combination with electronic structure calculations of NMR parameters, bond lengths, and bond angles. The computation of parameters from a statistical ensemble rather than from a single set of atomic Cartesian coordinates gives values that are in close quantitative agreement with the 2H NMR electric field gradient tensor measurements and are more consistent with the molecular symmetry revealed by the NMR spectra. PMID- 18407710 TI - Controlled formation of optically reflective and electrically conductive silvered surfaces on polyimide film via a direct ion-exchange self-metallization technique using silver ammonia complex cation as the precursor. AB - Double-surface-silvered polyimide films have been successfully fabricated using silver ammonia complex cation ([Ag(NH3)2]+) as the silver precursor and 3,3',4,4' benzophenonetetracarboxylic dianhydride/4,4'-oxidianile- (BTDA/ODA-) based poly(amic acid) (PAA) as the polyimide precursor via a direct ion-exchange self metallization technique. The process has been clarified to involve the loading of silver(I) into PAA via ion exchange, the thermally induced reduction of silver(I) to silver(0) and the concomitant imidization of PAA to polyimide upon thermal treatment, the subsequent silver-catalyzed and oxygen-assisted decomposition of the polyimide overlayer, and the self-accelerated aggregation of silver clusters on the film surface to produce well-defined surface silver layers. By employing [Ag(NH3)2]+ solution with a concentration of only 0.01 M and an ion-exchange time of no more than 10 min, the controlled formation of highly reflective and conductive silver surfaces upon thermal treatment at 300 degrees C for less than 4.5 h indicates that the present work provides an efficient route and an effacious silver species for polyimide surface metallization. Although the alkaline characteristics of [Ag(NH3)2]+ have a strong hydrolysis effect on the polyimide precursor chains, the final metallized films retain the key mechanical and thermal properties of the pure polyimide. Films were characterized by ATR FTIR, XPS, ICP-AES, SEM, TEM, DSC, TGA, reflectivity, conductivity, and mechanical measurements. PMID- 18407711 TI - Two-color two-photon excitation using femtosecond laser pulses. AB - The use of two-color two-photon (2c2p) excitation easily extends the wavelength range of Ti:sapphire lasers to the UV, widening the scope of its applications especially in biological sciences. We report observation of 2c2p excitation fluorescence of p-terphenyl (PTP), 2-methyl-5-t-butyl-p-quaterphenyl (DMQ) and tryptophan upon excitation with 400 and 800 nm wavelengths using the second harmonic and fundamental wavelength of a mode-locked Ti:sapphire femtosecond laser. This excitation is energetically equivalent to a one-photon excitation wavelength at 266 nm. The fluorescence signal is observed only when both wavelengths are spatially and temporally overlapping. Adjustment of the relative delay of the two laser pulses renders a cross correlation curve which is in good agreement with the pulse width of our laser. The fluorescence signal is linearly dependent on the intensity of each of the two colors but quadratically on the total incident illumination power of both colors. In fluorescence microscopy, the use of a combination of intense IR and low-intensity blue light as a substitute for UV light for excitation can have numerous advantages. Additionally, the effect of differently polarized excitation photons relative to each other is demonstrated. This offers information about different transition symmetries and yields deeper insight into the two-photon excitation process. PMID- 18407712 TI - Crystallization near glass transition: transition from diffusion-controlled to diffusionless crystal growth studied with seven polymorphs. AB - A remarkable property of certain glass-forming liquids is that a fast mode of crystal growth is activated near the glass transition temperature Tg and continues in the glassy state. This growth mode, termed GC (glass-crystal), is so fast that it is not limited by molecular diffusion in the bulk liquid. We have studied the GC mode by growing seven polymorphs from the liquid of ROY, currently the top system for the number of coexisting polymorphs of known structures. Some polymorphs did not show GC growth, while others did, with the latter having higher density and more isotropic molecular packing. The polymorphs not showing GC growth grew as compact spherulites at all temperatures; their growth rates near Tg decreased smoothly with falling temperature. The polymorphs showing GC growth changed growth morphologies with temperature, from faceted single crystals near the melting points, to fiber-like crystals near Tg, and to compact spherulites in the GC mode; in the GC mode, they grew at rates 3-4 orders of magnitude faster with activation energies 2-fold smaller than the polymorphs not showing GC growth. The GC mode had rates and activation energies similar to those of a polymorphic transformation observed near Tg. The GC mode was disrupted by the onset of the liquid's structural relaxation but could persist well above Tg (up to 1.15 Tg) in the form of fast-growing fibers. We consider various explanations for the GC mode and suggest that it is solid-state transformation enabled by local molecular motions native to the glassy state and disrupted by the liquid's structural relaxation (the alpha process). PMID- 18407713 TI - Stereoselective disposition of proton pump inhibitors. AB - It is estimated that about half of all therapeutic agents are chiral, but most of these drugs are administered in the form of the racemic mixture, i.e. a 50/50 mixture of its enantiomers. However, chirality is one of the main features of biology, and many of the processes essential for life are stereoselective, implying that two enantiomers may work differently from each other in a physiological environment. Thus, receptors or metabolizing enzymes would recognize one of the ligand enantiomers in favour of the other. With one exception, all presently marketed proton pump inhibitors (PPIs)--omeprazole, lansoprazole, pantoprazole and rabeprazole--used for the treatment of gastric acid-related diseases are racemic mixtures. The exception is esomeprazole, the S enantiomer of omeprazole, which is the only PPI developed as a single enantiomer drug. The development of esomeprazole (an alkaline salt thereof, e.g. magnesium or sodium) was based on unique metabolic properties that clearly differentiated esomeprazole from omeprazole, the racemate. At comparable doses, these properties led to several clinical advantages, for example higher bioavailability in the majority of patients, i.e. the extensive metabolizers (EMs; 97% in Caucasian and 80-85% in Asian populations), lower exposure in poor metabolizers (PMs; 3% in Caucasian and 15-20% in Asian populations) and lower interindividual variation. For the other, i.e. racemic, PPIs there are some data available on the characteristics of the individual enantiomers, and we have therefore undertaken to analyse the current literature with the purpose of evaluating the potential benefits of developing single enantiomer drugs from lansoprazole, pantoprazole and rabeprazole. For lansoprazole, the plasma concentrations of the S-enantiomer are lower than those of the R-enantiomer in both EMs and PMs, and, consequently, the variability in the population or between EMs and PMs is not likely to decrease with either of the lansoprazole enantiomers. Furthermore, plasma protein binding differs between the two lansoprazole enantiomers, in that the amount of the free S-enantiomer is two-fold higher than that of the R-enantiomer. This will counteract the difference seen in total plasma concentrations of the enantiomers. Also, studies using expressed human cytochrome P450 isoenzymes show that the metabolism of one enantiomer is significantly affected by the presence of the other, which is likely to result in different pharmacokinetics when administering a single enantiomer. For pantoprazole, there is a negligible difference in plasma concentrations between the two enantiomers in EMs, while the difference is substantial in PMs. The difference in AUC between PMs and EMs would decrease to some extent, but in the majority of the population the variability and efficacy would not be altered with a single enantiomer of pantoprazole. The metabolism of the enantiomers of rabeprazole displays stereoselectivity comparable to that of lansoprazole, i.e. the exposure of the R-enantiomer is higher than that of the S enantiomer in EMs as well as in PMs, which, by analogy to lansoprazole, makes them less suitable for development of a single enantiomer drug. Furthermore, the chiral stability of the rabeprazole enantiomers may be an issue because of significant degradation of rabeprazole to its sulfide analogue, which is subject to non-stereoselective metabolic regeneration of a mixture of the two enantiomers. In conclusion, in contrast to esomeprazole, the S-enantiomer of omeprazole, minimal if any clinical advantages would be expected in developing any of the enantiomers of lansoprazole, pantoprazole, or rabeprazole as compared with their racemates. PMID- 18407714 TI - Stroke risk screening of adults with hypertension: prospective cross-sectional study in primary care. AB - OBJECTIVES: There is a lack of current epidemiological data on the risk of cerebrovascular disease associated with arterial hypertension. We therefore aimed to assess the risk factor profile of hypertensive patients in primary care in various age groups, and to calculate their corresponding risk of stroke. METHODS: A total of 2482 primary-care practices throughout Germany included 47,394 consecutive unselected patients with diagnosed hypertension in a cross-sectional prospective observational study. In addition to demographic characteristics, participating primary-care physicians documented known risk factors for stroke using standardized questionnaires. The 10-year prospective risk of first stroke was then calculated according to the Framingham Stroke Risk Score. MAIN OUTCOME MEASURES: Patients were evenly balanced for sex (females 51%), and the mean age was 66.5 years. Mean systolic (SBP)/diastolic (DBP) blood pressure was 147/86 mmHg. Only 29.1% of patients had an SBP <140 mmHg, and 60.2% had a DBP <90 mmHg. The most prevalent risk factors were a positive family history of cardiovascular disease (46.1%), diabetes mellitus (36.1%), coronary artery disease (34.4%), and left ventricular hypertrophy (33.3%). Drug treatment was given as combination therapy in 73.5% of the total cohort of patients. The mean 10-year risk of stroke was 26% in the total cohort (0-19% in 50.6% of patients, 20-49% in 32.7%, and > or =50% in 16.7%). CONCLUSIONS: Co-morbidities relevant for total stroke risk are very prevalent among typical primary-care patients, confirming a substantial burden of disease in this setting. The resulting risk of stroke is substantial. The need for more aggressive BP control and treatment of modifiable risk factors is confirmed by our results. PMID- 18407715 TI - A markov model of the cost effectiveness of olanzapine treatment for agitation and psychosis in Alzheimer's disease. AB - BACKGROUND AND OBJECTIVE: Antipsychotics have long been used to treat agitation and psychosis related to Alzheimer's disease, but in a limited fashion because of troubling adverse effects. The new atypical antipsychotics are thought to be at least as effective as first-generation drugs and to cause fewer adverse effects. These drugs, however, are currently not US FDA-approved for use among elderly demented subjects due to a slight increase in the risk of death and serious cardiovascular events within this population. However, their favourable adverse effect profile has led many physicians to prescribe these drugs as first-line therapy for behaviourally disturbed patients with Alzheimer's disease. Clinical trials to evaluate the use of atypical antipsychotics have produced varying results, and clarity has not yet been achieved. Thus, a quantitative summary of the risks and benefits may help inform complex decisions that must be made in this area. In this study we set out to compare the expected costs and outcomes for a community-dwelling cohort of patients with Alzheimer's disease with agitation and/or psychosis who are (a) untreated, or (b) treated with olanzapine. METHODS: We constructed a Markov state-transition model using the best published data from several sources for Alzheimer's disease patient progression and treatment. This model allowed us to compare the expected costs and outcomes associated with olanzapine treatment compared with no treatment for a synthetic cohort of US adults aged > or =65 years. The model cycles every 6 months and continues until all patients die from Alzheimer's disease progression or from co morbid conditions. Outcome estimates included the incremental cost-effectiveness ratio (ICER) and cost per quality-adjusted life-year (QALY) gained. The robustness of the estimates was examined by sensitivity analyses of key parameters, including cost of care, olanzapine effectiveness and Alzheimer's disease progression rates. RESULTS: Results indicated that olanzapine was a cost effective treatment for agitation and psychosis related to Alzheimer's disease when compared with no treatment (ICER <$US50,000). In addition, sensitivity analyses demonstrated that olanzapine remained cost effective despite multiple variations of several parameters, both alone and concurrently. CONCLUSION: Olanzapine treatment for agitation and psychosis related to Alzheimer's disease is cost effective when compared with no treatment. Further analysis should be performed as atypical antipsychotics become generic, as more information on health utilities in the Alzheimer's disease population becomes available, and to compare atypical antipsychotics with first-generation antipsychotics. PMID- 18407716 TI - Does iatrogenic scleroderma due to injection-site reaction to enfuvirtide impair absorption of the drug? AB - BACKGROUND: Chronic iatrogenic scleroderma is a possible obstacle to the absorption of subcutaneously administered drugs. This study correlated the clinical and histopathological pattern of injection-site reactions (ISRs) to the pharmacokinetics of enfuvirtide in patients with HIV. METHODS: Fourteen patients treated with an enfuvirtide-based antiretroviral regimen for a median of 45 weeks were enrolled and their ISRs were evaluated. Twelve patients with evidence of ISRs underwent cutaneous biopsies using a 4-mm punch. The maximum plasma enfuvirtide concentration (Cmax) and the area under the enfuvirtide concentration time curve (AUC) were assessed using blood sampling. RESULTS: Four different macroscopic patterns of ISR were identified: A--no evidence of cutaneous lesions; B--transient infiltrative lesions that auto-resolved within 24 hours; C- transient nodular lesions that auto-resolved within 7-15 days; and D--stable lesions after more than 30 days. Histological examination showed three morphological patterns: (1) acute urticaria/vasculitis-like pattern, (2) subacute pattern and (3) chronic scleroderma-like pattern. No differences among patients with the various patterns of ISRs were observed, except for a higher Cmax and AUC in patients with pattern 1. CONCLUSIONS: These results confirm that although iatrogenic scleroderma is not related to impaired enfuvirtide absorption, higher Cmax and AUC values are observed in patients with urticaria/vasculitis-like patterns. PMID- 18407717 TI - Efficacy, safety and pharmacokinetic study of a novel fentanyl-containing matrix transdermal patch system in Japanese patients with cancer pain. AB - BACKGROUND AND OBJECTIVES: A novel transdermal matrix patch delivery system for fentanyl has been developed to deliver improved management of cancer pain compared with that obtained using current fentanyl reservoir patches. This study was carried out to assess the efficacy, safety and pharmacokinetic profiles of a 12.5 microg/h transdermal matrix fentanyl patch administered with the objective of replacing morphine, oral oxycodone or fentanyl injection formulations. The study also evaluated how the pharmacokinetic profiles of higher dose fentanyl patches (25, 37.5 and 50 microg/h) changed following dose adjustments to optimize management of cancer pain. METHODS: This open-label, multicentre study involved 87 patients of both sexes (> or =20 years) with a confirmed diagnosis of cancer. Patients were receiving any one of the following at the time of enrollment for the management of their cancer pain: (a) morphine <45 mg/day orally, <30 mg/day as suppositories, or <15 mg/day by injection; (b) oral oxycodone <30 mg/day; or (c) fentanyl injectable preparations <0.3 mg/day. The patients were administered a 3-day course of fentanyl transdermal matrix patch application three times. The initial dose was 12.5 microg/h, which could be increased when a new patch was applied if the physician deemed this to be appropriate based on pain intensity ratings and use of rescue medications. Efficacy outcomes included patients' global assessment scores (primary efficacy endpoint) measured on a five-step scale and dichotomous scores for physicians' global assessment. The occurrence of adverse events and changes in laboratory tests were evaluated as safety variables. Serum fentanyl levels were measured immediately after removal of the old patch on days 4, 7 and 10 to obtain data on trough serum concentrations. RESULTS: The percentage of patients in category 3 or higher (very satisfied, satisfied, or neither satisfied nor dissatisfied) for the patient's global assessment score was 89.4% (76/85), indicating high patient satisfaction and attainment of sufficient pain control after patients switched from their previously used opioid analgesics. Similar findings were obtained on physicians' global assessment scores. A total of 316 adverse events occurred in 78 (90.7%) of 86 patients who were administered at least one patch. These included nausea (31 [36.0%]), somnolence (26 [30.2%]), vomiting (22 [25.6%]), diarrhoea (17 [19.8%]), constipation (14 [16.3%]), pyrexia (11 [12.8%]) and insomnia (9 [10.5%]). The mean (+/- SD) serum fentanyl concentration determined on day 4 was 169.9 +/- 103.4 pg/mL (n = 83). Serum fentanyl measurement results indicated that the same fentanyl patch dose resulted in similar serum fentanyl levels, while increased doses produced higher serum fentanyl concentrations. CONCLUSION: The fentanyl matrix transdermal patch formulation employed in this study demonstrated sufficient cancer pain control for patients switching from morphine or oral oxycodone preparations. The patch tested was well tolerated and its use did not result in any increased incidence of adverse drug reactions over those commonly found with opioid analgesics. PMID- 18407718 TI - Retrospective analysis of rosiglitazone and macular oedema in patients with type 2 diabetes mellitus. AB - BACKGROUND: Macular oedema tends to be a more rapid complication of diabetic retinopathy and represents the major cause of blindness. Among subjects with type 2 diabetes mellitus, it can be found in 15% of those who use insulin and 4% of those who do not. Use of thiazolidinediones (glitazones) has recently been associated with some cases of macular oedema. METHODS: We recalled 102 diabetic subjects treated with rosiglitazone to our diabetes centre in order to evaluate a possible association of this drug with macular oedema. Of these 102 subjects, we evaluated all 76 who provided written informed consent to participate in the analysis. All of these underwent a battery of four diagnostic tests: (1) visual acuity, (2) Amsler visual field test, (3) Ishihara colour recognition test, and (4) retinal fundus photography. All retinal photographs were examined by two experienced ophthalmologists. RESULTS: The most noticeable result was that most subjects (80%) had satisfactory visual acuity. The Ishihara test chart showed that three subjects were colour blind, but this abnormality was already known. On the Amsler test, one subject had a positive result consisting of visual distortion of a series of straight lines. In the retinal photos, two expert ophthalmologists independently identified one case of 'paramacular oedema' in a subject with diabetes of long duration with a proliferative retinopathy. The patient developed bilateral macular oedema during treatment with rosiglitazone 8 mg/day. The patient had been diagnosed with diabetes at the age of 45 years and after a period of 6 years taking oral antihyperglycaemic agents had been switched to insulin, up to four injections per day (total 60-70 IU/day), for the next 15 years. In 2000 a routine examination demonstrated the presence of sustained hypertension and the patient was started on an ACE inhibitor. A computerized test for autonomic neuropathy demonstrated abnormal deep breathing and lying-to standing responses. Treatment with rosiglitazone was interrupted and the subject underwent a series of retinal photocoagulations for proliferative retinopathy. Two months after rosiglitazone therapy had been discontinued, the visual acuity of the patient reversed to baseline values. CONCLUSION: The study shows that rosiglitazone was not linked to formation of macular oedema, with the exception of one case of bilateral and clinically reversible paramacular oedema, where rosiglitazone was given in co-administration with a long-term insulin treatment regimen in a subject with pre-existing diabetic retinopathy. This patient had a long duration of diabetes and had also been hypertensive since 2000. PMID- 18407720 TI - Multigroup confirmatory factor analysis for the adaptive behavior assessment system-II parent form, ages 5-21. AB - The AAIDD has promulgated various models of adaptive behavior, including its 1992 model stressing 10 adaptive skills and its 2002 model that highlighted three conceptual domains. In previous studies on the Adaptive Behavior Assessment System-II (ABAS-II), researchers found support for a model including both 10 adaptive skills and three conceptual domains. To extend this review, we examined gender-invariant structure of adaptive behavior using the ABAS-II Parent Form, Ages 5-21, to answer four questions: Do the skill areas in this measure display the same pattern of factor loadings and the same factor loadings? Are intercepts of the observed skill areas equal? Do skill areas measure the corresponding factors with the same accuracy? Results show a similar one-factor structure for males and females. PMID- 18407719 TI - Perceived quality of life in mothers of children with fragile X syndrome. AB - A mixed method approach was used to explore quality of life of 101 mothers of children with fragile X syndrome. Mothers completed a self-report of personal quality of life and measures of mental health and well-being. A subset was interviewed about quality of life. The distribution of scores on the Quality of Life Inventory was similar to the norms, indicating that these mothers do not, as a whole, report a lower quality of life than does the average woman. Significant predictors of quality of life were trait hope and parenting stress. The most common positive factor was being a mother. However, they also indicated that parenting a child with fragile X presented challenges and stressors, primarily when the social environment was not supportive. PMID- 18407721 TI - Short-term memory coding in children with intellectual disabilities. AB - To examine visual and verbal coding strategies, I asked children with intellectual disabilities and peers matched for MA and CA to perform picture memory span tasks with phonologically similar, visually similar, long, or nonsimilar named items. The CA group showed effects consistent with advanced verbal memory coding (phonological similarity and word length effects). Neither the intellectual disabilities nor MA groups showed evidence for memory coding strategies. However, children in these groups with MAs above 6 years showed significant visual similarity and word length effects, broadly consistent with an intermediate stage of dual visual and verbal coding. These results suggest that developmental progressions in memory coding strategies are independent of intellectual disabilities status and consistent with MA. PMID- 18407722 TI - Veridical and false pictorial memory in individuals with and without mental retardation. AB - Veridical and false pictorial recognition were assessed in individuals with mental retardation; groups were matched for MA and CA. Pictures were viewed in either a generative or static format at acquisition. The individuals with mental retardation and those in the MA-matched group had higher rates of false memories for critical items and lower hit rates than did their CA-matched peers. The mental retardation group demonstrated an acquiescent response bias (i.e., high novel false-alarm rate). When data were corrected for this bias, those with mental retardation had significantly lower hit rates but equivalent false-alarm rates to the MA-matched participants. Results are discussed in terms of pictorial distinctiveness and within the frameworks of activation monitoring and fuzzy trace theory. PMID- 18407724 TI - Interrater reliability of the Supports Intensity Scale (SIS). AB - The interrater reliability of the Supports Intensity Scale (SIS) was investigated under the condition that interviewers had to have been trained and/or experienced in its administration and scoring. Both corrected and noncorrected Pearson's product-moment coefficients were generated to assess interinterviewer, interrespondent, and mixed interrater reliability. The correlation coefficients for the SIS Support Needs Index Score and SIS subscale scores were considerably higher than coefficients reported in the SIS Users Manual that were derived from a similar study conducted with untrained, inexperienced interviewers. PMID- 18407727 TI - Linking fatherhood to prostate cancer risk. PMID- 18407723 TI - Signaling noncomprehension of language: a comparison of fragile X syndrome and Down syndrome. AB - Signaling noncomprehension of the spoken messages of others was examined for youth with fragile X or Down syndrome in comparison with each other and nonverbal MA-matched typically developing children. A direction-following task was used in which some of the directions were inadequate. Both syndrome groups signaled noncomprehension less often than did the typically developing children. The ability to signal noncomprehension appropriately was related to a measure of receptive vocabulary and syntax. Preliminary analyses indicated that males with fragile X syndrome signaled noncomprehension less often than did their female peers, even after controlling for differences in nonverbal MA. PMID- 18407728 TI - Medical intensive care for lung cancer patients: better than we thought? PMID- 18407730 TI - Bortezomib in mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) represents 6% of non-Hodgkin lymphomas, but is one of the most active fields of clinical investigation. Unfortunately, there is still no standard or curative therapy in MCL. Front-line therapy appears to benefit from intensification either through high-dose therapy with stem cell transplant consolidation or dose-intense chemotherapy with hyperfractionated cyclophosphamide, vincristine, adriamycin/doxorubicin and dexamethasone/rituximab. Most patients still relapse and a multitude of novel agents are currently being tested in this setting, including proteasome inhibitors with bortezomib (the first of its class and the first US FDA-approved drug for MCL), mTOR inhibitors, Bcl-2 inhibitors, antiangiogenesis agents and histone deacetylase inhibitors among others. An obvious effort is needed to enroll patients on clinical trials, the design of which might benefit from pharmacogenomics and a better understanding of MCL biology and its diversity. PMID- 18407731 TI - Histamine dihydrochloride for the treatment of acute myeloid leukemia, malignant melanoma and renal cell carcinoma. AB - Histamine dihydrochloride is a vasoactive biogenic amine. It inhibits the reactive oxygen species formation in monocytes via histamine H2 receptors and protects natural killer and T cells from oxidative damage. Histamine has the potential to optimize cytokine-induced activation of T cells and natural killer cells; therefore, the addition of histamine to cytokine treatment may improve treatment efficacy. Clinical trials in solid tumors and in acute myeloid leukemia have demonstrated the potential to improve treatment outcome when histamine dihydrochloride is combined with immunotherapy. In patients with metastatic malignant melanoma, this strategy improved remission rates and increased survival. On the other hand, less promising results were reported for histamine dihydrochloride added to cytokines in patients with other solid tumors, especially in advanced renal cell carcinoma. A recent international Phase III trial performed in 320 patients showed that maintenance therapy with histamine dihydrochloride and IL-2 was able to improve leukemia-free survival in patients with acute myeloid leukemia, without an effect on overall survival. The combination of histamine dihydrochloride with IL-2 potentially offers an efficacious and tolerable maintenance strategy for patients with acute myeloid leukemia; however, its impact on survival remains to be explored. PMID- 18407732 TI - Capecitabine in the treatment of advanced gastric cancer. AB - Although the role of systemic chemotherapy has been established for the treatment of advanced gastric cancer, the prognosis of these patients remains poor, with a median overall survival of less than 1 year. Based on the results of several randomized Phase III trials, 5-fluorouracil continuous infusion plus cisplatin, with or without epirubicin, has become the global reference regimen for this patient population. However, treatment with fluorouracil infusion requires either frequent hospitalizations or the use of a central venous access device, harboring potential complications. Capecitabine, a tumor-activating oral prodrug of fluorouracil, may be more advantageous in terms of patient convenience, safety and efficacy. Two recent randomized Phase III trials have shown that capecitabine could replace infusional fluorouracil in cisplatin-based regimens. Furthermore, Phase II trials have shown that many other capecitabine-based doublet or triplet chemotherapy regimens incorporating newer cytotoxic agents are active and well tolerated. Many promising biological agents are now being tested in Phase III trials, incorporating capecitabine combinations as control arms, in patients with advanced gastric cancer. PMID- 18407733 TI - Front-line therapy for nonlocalized diffuse large B-cell lymphoma: what has been demonstrated and what is yet to be established. AB - The field of treatment of diffuse large B-cell lymphoma has been in a continuous flux over the last 10-15 years owing to the introduction of new therapeutic approaches such as dose-dense chemotherapy, monoclonal antibodies and high-dose chemotherapy followed by autologous peripheral blood stem cell transplant. The use of clinical prognostic factors has improved our ability to predict the outcome of these lymphomas; moreover, the gene and protein expression pattern has been shown, at least in the pre-rituximab era, to be an independent and powerful prognostic indicator. This review will focus on results obtained in the last decade by large clinical trials evaluating the first-line therapy in nonlocalized diffuse large B-cell lymphoma; special emphasis will be placed on more mature results that can be indicated as 'standard' therapy. Ongoing studies addressing as yet unanswered or controversial questions will be analyzed, and preliminary data will be critically reviewed. PMID- 18407734 TI - Imatinib as effective therapy for dermatofibrosarcoma protuberans: proof of concept of the autocrine hypothesis for cancer. AB - Cancer literature has consistently described autocrine loops involving growth factors to be important mechanisms for cellular transformation and proliferation in preclinical cancer models. Finally, convincing clinical data exist to implicate autocrine loops as central to the pathogenesis of a malignant condition, largely as a result of the recent development of inhibitors of protein tyrosine kinases important in cell signaling and growth. Although a rare condition, the study of patients with dermatofibrosarcoma protuberans (DFSP) is enriched by data demonstrating strong scientific rationale for its pathogenesis and susceptibility to molecular-based therapies. DFSP is a low-grade sarcoma that responds to treatment with imatinib, an inhibitor of the PDGF receptor tyrosine kinase. This treatment response illustrates the importance of autocrine/paracrine loops involving PDGF and its receptor as the key molecular target in DFSP. New insight into this fundamental biological mechanism sets the scene for the further development of molecular-targeted therapeutic options for cancer. PMID- 18407735 TI - Translational research in glioblastoma multiforme: molecular criteria for patient selection. AB - In spite of the dismal outcome of glioblastoma multiforme (GBM), we are in a position to provide a ray of hope to patients and families. Methylation of MGMT in tumor occurs in approximately a third of patients and predicts meaningful response and survival to adjuvant radiotherapy plus temozolomide. Limited access to tumor tissue in some patients could be circumvented by examining MGMT methylation in circulating serum DNA, although this approach needs to be validated. Molecular signatures are also promising prognostic and predictive markers, and clinical trials should be carried out to validate their use in the selection of patients for specific targeted therapies. Gene expression by quantitative PCR of key components of these molecular signatures could pave the way for easy identification of different subgroups of patients. Translational clinical trials are warranted in order to detect the subgroups of patients resistant to radiotherapy who may derive benefit from novel therapies, including antiangiogenic drugs. PMID- 18407736 TI - Hereditary diffuse gastric cancer and lost cell polarity: a short path to cancer. AB - The mechanisms that underlie the initiation of human cancer are poorly understood. Here, we describe the development of hereditary diffuse gastric cancer and argue that it arises from the disruption of the regenerative processes that are inherent to all epithelial tissues. This model supports the cancer stem cell hypothesis, in which tumors contain a subpopulation of cells with the key stem cell characteristics of capacity for self renewal, differentiation and limitless replication. We argue that epigenetic modifications induced by common environmental and physiological pressures are able to initiate this disruption. The carcinogenic effects of these modifications are potentially reversible through the use of epigenetic therapies such as DNA demethylating agents and histone deacetylation inhibitors. PMID- 18407737 TI - Is there a role for advanced radiation therapy technologies in the treatment of pancreatic adenocarcinoma? AB - Pancreatic cancer remains a highly challenging problem in oncology. Oncologists continue to search for therapies that are more effective than those currently available to improve on the existing poor treatment results. Persistence of both systemic and local disease causes high rates of morbidity and mortality for patients. Radiation continues to play a role in the treatment of pancreatic cancer, in both the adjuvant and locally advanced settings. Efforts to improve on the results of radiotherapy have led to the use of new and improved technologies. This review discusses a variety of these technological improvements and their current and potential future roles in the clinic. PMID- 18407738 TI - Percutaneous renal cryoablation: current status. AB - Over the last 13 years, renal cryoablation has emerged as a promising technique for the treatment of solid renal tumors. The improvement in imaging modalities such as ultrasound, computed tomography and MRI, as well as the introduction of thinner probes, has led to the spread of the minimally invasive percutaneous approach. We review the historical background of percutaneous renal cryoablation (PRC), present its basic principles, mention the contemporary clinical data and outcomes of this technique and suggest future directions for its wider application in renal tumors. Early results have demonstrated that it may offer an alternative for the treatment of small renal masses with the advantages of minimal complications, spared renal function, decreased overall costs and equivalent oncologic efficacy. Long-term results are required in order to apply this minimally invasive technique to a broader spectrum of patients. PMID- 18407739 TI - Clinical proteomics and breast cancer: strategies for diagnostic and therapeutic biomarker discovery. AB - A major challenge of breast cancer research is the identification of accurate biomarkers that improve screening, early diagnosis, prediction of aggressiveness, and prediction of therapeutic response or toxicity, as well as the identification of new molecular therapeutic targets. The new proteomic techniques promise to be valuable for identifying such tissue and serum markers. The different techniques currently applied to clinical samples of breast cancer and the most important results obtained are summarized in this review. PMID- 18407740 TI - miRNA modulation of the cellular stress response. AB - Cellular stress responses are potent and dynamic, allowing cells to effectively counteract diverse stresses. These pathways are crucial not only for maintaining normal cellular homeostasis, but also for protecting cells from what would otherwise lead to their demise. A novel class of genes, termed miRNAs, has recently been implicated in the cellular stress response. For example, it has been demonstrated that a cardiac-specific miRNA that is not required for normal development is requisite for a normal cardiac stress response in mice. In addition, we have found that a miRNA family is able to modulate the cellular response to cytotoxic cancer treatment both in vitro and in vivo. In this review, we will discuss these and other important developments in the field. In particular, we will focus on studies that have linked miRNAs to the genotoxic stress response and will suggest how this connection may be both important for our understanding of biology and pertinent for the development of novel cancer therapies. PMID- 18407741 TI - Novel chemotactic-antigen DNA vaccine against cancer. AB - Dendritic cells play a pivotal role in immune induction. Dendritic cells perform antigen uptake, processing and presentation to T cells only when they are matured and in the functional state. In the development of a vaccine, it is of utmost importance to consider how to make dendritic cells' functions immunologically adequate. In this paper, we report the development of a series of antitumor DNA vaccines with similar structural framework, in which a gene encoding tumor associated antigenic peptide is ligated upstream to the gene coding secondary lymphoid-tissue chemokine and downstream to the gene encoding the Fc portion of IgG (named chemotactic-antigen DNA vaccine [CADV]). CCR7(+) T, B, natural killer and dendritic cells can be attracted by secondary lymphoid-tissue chemokine, and Fc facilitates antigen uptake via Fc receptors expressed on dendritic cells. In a series of experiments in mice vaccinated by CADV with such tumor-associated antigenic specificities as HPV-16 E7, PSA-PSM-PAP, HER-2/neu, p53 and hTERT, CADV can attract immune cells to the vaccine inoculation site, remarkably inhibit tumor growth and extend survival time in tumor-bearing mice. The antitumor effect is more efficacious than that in mice treated with SLC-Ag or Ag-Fc hybrid gene. Tumor-associated antigenic-specific cytotoxic T lymphocytes can be induced by in vitro experiment in a human system. When combined with measures blocking the negative immune feedback circuits, the therapeutic effect of the vaccine can be further enhanced. Large-scale production of CADV is possible for clinical application. PMID- 18407743 TI - Nutritional implications of genetic taste variation: the role of PROP sensitivity and other taste phenotypes. AB - Genetic sensitivity to the bitter taste of phenylthiocarbamide and 6-n propylthiouracil (PROP) is a well-studied human trait. It has been hypothesized that this phenotype is a marker for individual differences in taste perception that influence food preferences and dietary behavior with subsequent links to body weight and chronic disease risk. Steady progress has been made over the past several decades in defining the involvement of this phenotype and its underlying gene, TAS2R38, in this complex behavioral pathway. However, more work needs to be done to fully determine its overall nutritional and health significance. The primary goal of this review is to assess our current understanding of the role of the PROP bitter taste phenotype in food selection and body weight in both children and adults. A brief history of the field is included and controversies surrounding the use of different PROP screening methods are addressed. The contribution of other receptors (both bitter and nonbitter) to human taste variation is also discussed. PMID- 18407744 TI - The efficiency of cellular energy transduction and its implications for obesity. AB - We assess the existence, mechanism, and functions of less-than-maximal coupling efficiency of mitochondrial oxidative phosphorylation and its potential as a target for future antiobesity interventions. Coupling efficiency is the proportion of oxygen consumption used to make adenosine triphosphate (ATP) and do useful work. High coupling efficiency may lead to fat deposition; low coupling efficiency to a decrease in fat stores. We review obligatory and facultative energy expenditure and the role of a futile cycle of proton pumping and proton leak across the mitochondrial inner membrane in dissipating energy. Basal proton conductance is catalyzed primarily by the adenine nucleotide translocase but can be mimicked by chemical uncouplers. Inducible proton conductance is catalyzed by specific uncoupling proteins. We discuss the opportunities and pitfalls of targeting these processes as a treatment for obesity by decreasing coupling efficiency and increasing energy expenditure, either directly or through central mechanisms of energy homeostasis. PMID- 18407745 TI - Defining linkages between the GSC and NSF's LTER program: how the Ecological Metadata Language (EML) relates to GCDML and other outcomes. AB - The Genomic Standards Consortium (GSC) invited a representative of the Long-Term Ecological Research (LTER) to its fifth workshop to present the Ecological Metadata Language (EML) metadata standard and its relationship to the Minimum Information about a Genome/Metagenome Sequence (MIGS/MIMS) and its implementation, the Genomic Contextual Data Markup Language (GCDML). The LTER is one of the top National Science Foundation (NSF) programs in biology since 1980, representing diverse ecosystems and creating long-term, interdisciplinary research, synthesis of information, and theory. The adoption of EML as the LTER network standard has been key to build network synthesis architectures based on high-quality standardized metadata. EML is the NSF-recognized metadata standard for LTER, and EML is a criteria used to review the LTER program progress. At the workshop, a potential crosswalk between the GCDML and EML was explored. Also, collaboration between the LTER and GSC developers was proposed to join efforts toward a common metadata cataloging designer's tool. The community adoption success of a metadata standard depends, among other factors, on the tools and trainings developed to use the standard. LTER's experience in embracing EML may help GSC to achieve similar success. A possible collaboration between LTER and GSC to provide training opportunities for GCDML and the associated tools is being explored. Finally, LTER is investigating EML enhancements to better accommodate genomics data, possibly integrating the GCDML schema into EML. All these action items have been accepted by the LTER contingent, and further collaboration between the GSC and LTER is expected. PMID- 18407746 TI - Molecular genetic testing in the United States: comparison with international practice. AB - OBJECTIVE: To compare data on the practices of molecular genetic testing (MGT) in laboratories in the United States with those in 18 other countries. METHODS: A Web-based survey of MGT laboratory directors (n = 827; response rate 63%) in 18 countries on three continents was carried out, and the response from U.S. laboratories compared to all others. Quality assurance and reporting indices were developed and calculated for each responding laboratory. RESULTS: A comparison of U.S. results with all other countries identified differences in laboratory setting, personnel qualifications, and the specific tests being offered, but similar rates of adherence to MGT quality standards and reporting practices were found. The survey also documented substantial transborder flow of specimens, most commonly due to the lack of availability of the test in the United States or because the test was available only through a research protocol, highlighting the need for common reporting and practice guidelines for the international MGT community. CONCLUSION: The findings presented here provide further support for the need to consider the application of the Organisation for Economic Cooperation and Development (OECD) Guidelines and the establishment of compatible accreditation programs or equivalent mechanisms across national borders to ensure the quality of laboratory services and the clinical usefulness of molecular genetic test reports for referred specimens. PMID- 18407747 TI - Muscular metabolism in aged rats under exhaustive exercise: effect of a modified alkaline supplementation. AB - A modified alkalizing supplementation (MAS) was tested on skeletal muscle metabolism in aged rats undergoing exhaustive exercise. Aged Wistar rats were allocated into two groups: saline (A) and saline added with 16 mg of MAS (B) before treadmill exercise. Blood and gastrocnemius and soleus muscle were analyzed after exercise for succinate dehydrogenase (SDH), acetylcarnitine (ALCAR), and glycogen. Lactic acid (LA), creatin-phosphokinase (CPK), and gas analysis were tested in the blood. Exercise caused a significant increase of LA and CPK and muscle glycogen fall. Arterial desaturation at exhaustion was prevented in the B group (p < 0.05). Exercise-induced increase of SDH and ALCAR was further enhanced in B rats (p < 0.05). This study suggests that MAS can improve fast and endurance muscle metabolism in aged rats by increasing cellular acetyl group availability and tricarboxylic acid turnover. PMID- 18407748 TI - Adeno-sh-beta-catenin abolishes ischemic preconditioning-mediated cardioprotection by downregulation of its target genes VEGF, Bcl-2, and survivin in ischemic rat myocardium. AB - beta-Catenin, the downstream target of glycogen synthase kinase-3beta (GSK 3beta), plays a vital role in ischemic preconditioning (IP)-mediated cardioprotection. In the present study, we investigated the mechanism of IP mediated cardioprotection through suppression of beta-catenin expression by intramyocardial injection of adeno-sh-RNA against beta-catenin (BCT) (4 x 10(8) pfu). Adeno-LacZ (LZ) was used as control. The rats were randomized into (a) LZ + ischemia-reperfusion (IR); (b) LZIPIR; (c) BCTIR; and (d) BCTIPIR. Isolated hearts from each group were subjected to 30 min of I followed by 2 h of R. Both IPIR group hearts were subjected to IP (5 min I + 10 min R; four cycles) before IR. Significant reduction in left ventricular functional recovery (78 vs. 88 mm Hg), dp/dt(max) (1,802 vs. 2,189 mm Hg/sec), and aortic flow (4 vs. 9 ml/min) was observed in BCTIPIR compared with LZIPIR at 120 min of reperfusion. Increased infarct size (42 vs. 24%) and apoptotic cardiomyocytes (122 vs. 58 counts/60 HPF) were observed in BCTIPIR compared with LZIPIR. Realtime PCR and Western blot analysis showed significant downregulation in mRNA and protein expression of VEGF, Bcl-2, and survivin in BCTIPIR compared with LZIPIR. These findings indicated for the first time that silencing beta-catenin abolished IP-mediated cardioprotection, probably through inhibition of VEGF-Bcl-2 and survivin. PMID- 18407749 TI - In situ detection and measurement of intracellular reactive oxygen species in single isolated mature skeletal muscle fibers by real time fluorescence microscopy. AB - Reactive oxygen species (ROS) produced by skeletal muscle stimulate adaptive responses to activity and mediate some degenerative processes. ROS activity is usually studied by measuring indirect end-points of their reactions with various biomolecules. In order to develop a method to measure the intracellular ROS generation in real-time in mature skeletal muscle fibers, these were isolated from the flexor digitorum brevis (FDB) muscle of mice and cultured on collagen coated plates. Fibers were loaded with 5- (and 6-) chloromethyl-2',7' dichlorodihydrofluorescein diacetate (CM-DCFH DA) and measurements of 5- (and 6-) chloromethyl-2',7'-dichlorofluorescin (CM-DCF) fluorescence from individual fibers obtained by microscopy over 45 min. The sensitivity of this approach was demonstrated by addition of 1 microM H(2)O(2) to the extracellular medium. Contractions of isolated fibers induced by field electrical stimulation caused a significant increase in CM-DCF fluorescence that was abolished by pre-treatment of fibers with glutathione ethyl ester. Thus, CM-DCF fluorescence microscopy can detect physiologically relevant changes in intracellular ROS activity in single isolated mature skeletal muscle fibers in real-time, and contractions generated a net increase that was abolished when the intracellular glutathione content was enhanced. This technique has advantages over previous approaches because of the maturity of the fibers and the analysis of single cells, which prevent contributions from nonmuscle cells. PMID- 18407750 TI - Editorial and commentary on the guidance document for the contained field trials of engineered mosquitoes. PMID- 18407752 TI - Differential behavior of auricular and articular chondrocytes in hyaluronic acid hydrogels. AB - Chondrocytes isolated from a variety of sources, including auricular (AU) and articular (AR) cartilage, can differ in cell behavior, growth, and extracellular matrix (ECM) production, which can impact neocartilage properties in tissue engineering approaches. This behavior is also affected by the surrounding microenvironment, including soluble factors, biomaterials, and mechanical loading. The objective of this study was to investigate differences in juvenile AU and AR chondrocyte behavior when encapsulated in radically polymerized hyaluronic acid hydrogels. When implanted in vivo, differences in macroscopic appearance, mechanical properties, glycosaminoglycan content, and collagen content were observed depending on the chondrocyte type encapsulated. Specifically, AU constructs exhibited construct growth and neocartilage formation with increases in aggregate modulus and ECM accumulation with culture, whereas AR constructs retained their construct size and remained translucent with only a minimal increase in the compressive modulus. When cultured in vitro, both cell types remained viable and differences in gene expression were observed for type I and II collagens. Likewise, differences in gene expression were noted after dynamic mechanical loading, where stimulated AR constructs exhibited 2.3- and 1.5 fold increases in type II collagen and aggrecan over free-swelling controls, while AU samples exhibited smaller fold increases of 1.4- and 1.3-fold, respectively. Thus, these data indicate that the specific cell source, cell/material interactions, and loading environment are important in the final properties of tissue-engineered products. PMID- 18407753 TI - Is sleeping position related to asymmetry in bilateral Graves' ophthalmopathy? AB - BACKGROUND: In agreement with the systemic nature of Graves' disease, Graves' ophthalmopathy (GO) presents as a symmetric bilateral eye disease in the vast majority of patients. However, asymmetric involvement of both eyes is frequently observed. We hypothesized that sleeping position might be involved in asymmetric GO; when, for example, the preferred sleeping position is on the right side, retrobulbar pressure might be somewhat higher in the right than in the left orbit, resulting in more severe eye changes in the right eye. METHODS: A prospective study in 75 consecutive untreated patients with GO, in whom eye changes and sleeping position were assessed independent of each other. Criteria for asymmetric eye changes were differences between both eyes in the presence of retrobulbar pain or of > or =1 grade in soft tissue involvement, and/or of > or =2 mm in exophthalmos, and/or > or =8 degrees in elevation. Preferred sleeping position was determined by questionnaire. RESULTS: All patients had bilateral GO. Eye changes were symmetrical in 50 patients, and asymmetrical in 25 patients based on differences in proptosis and elevation. Sleeping position did not differ between symmetric and asymmetric GO. Preferred sleeping position was on the left side in 23%; on the right side in 31%; and on the back, on the belly, or unknown in 46%. In patients with asymmetric GO, the most affected ("worst") eye was not related to preferred sleeping position on the right or left side. CONCLUSION: Slight asymmetry of eye changes between both eyes in patients with bilateral GO is not related to preferred sleeping position on either the right or left side. PMID- 18407754 TI - Huge thyroid uptake of 18F-FDG in a patient with Hashimoto's thyroiditis referred for a malignant thyroid lesion. PMID- 18407755 TI - Palpation thyroiditis causing new-onset atrial fibrillation. AB - OBJECTIVE: Surgery-induced thyroiditis can pose a significant clinical problem that is underappreciated. We present a case of new-onset atrial fibrillation as a consequence of palpation thyroiditis in a 70-year-old man who underwent radical right neck dissection for malignant melanoma and review the limited literature on this topic. DESIGN: Biochemical parameters including thyrotropin, free thyroxine, triiodothyronine, erythrocyte sedimentation rate, C-reactive protein, thyroglobulin, thyroid stimulating immunoglobulins, thyroid binding inhibitory immunoglobulins, thyroid peroxidase, thyroglobulin antibodies, and 24-hour urine iodine were measured. Thyroid ultrasound and technetium-99m pertechnetate scintigraphy with radioactive iodine 131 uptake were employed for diagnostic purposes. MAIN OUTCOME: Following right neck exploratory dissection, the patient developed hyperthyroidism and atrial fibrillation. Imaging studies were compatible with right lobar thyroiditis. Other etiologies of thyroiditis were excluded. Despite normalization of thyroid function after 2 weeks, atrial fibrillation persisted and required cardioversion. CONCLUSIONS: Manipulation of the thyroid gland during neck exploratory surgery can result in hyperthyroidism with atrial fibrillation as a consequence. To avoid this complication, careful attention should be paid during surgical procedures or other clinical situations in which the thyroid gland is manipulated. PMID- 18407756 TI - The role of ultrasound in thyroid nodules with a cytology reading of "suspicious for papillary thyroid carcinoma". AB - BACKGROUND: Irrespective of ultrasound (US) features, surgery is usually recommended for patients who have a fine-needle aspiration biopsy (FNAB) read as suspicious for papillary carcinoma (PTC). The aim of the present study was to evaluate the role of US in the management of thyroid nodules with a FNAB reading suspicious for PTC. METHODS: Between August 2002 and May 2006, 303 patients who had thyroid nodules with a FNAB reading suspicious for PTC underwent surgery. The sonographic findings in the patients were classified as suspicious for malignancy or probably benign based on the US reading. The US readings and final pathological diagnoses of thyroid nodules were analyzed in these patients. RESULTS: The malignancy rate was 84.2% in patients with a FNAB specimen suspicious for PTC. Sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of the US were 96.4%, 74.5%, 92.7%, 94.9%, and 80.9%, respectively, in nodules read as suspicious for PTC on FNAB. Whereas 243 (96.4%) lesions were found to be malignant at surgery, in the 252 lesions that had ultrasound findings suspicious for malignancy, only 13 (25.5%) lesions were malignant out of the 51 that had US readings of probably benign (p < 0.05). CONCLUSIONS: The probability of malignancy is much lower in thyroid nodules with benign US findings even if the FNAB is read as suspicious for PTC. Therefore, US may be useful in planning the extent of surgery in patients with a FNAB reading of suspicious for PTC. As thyroid malignancy occurs in approximately 26% of patients with cytology readings suspicious for PTC and benign-appearing US, the US reading alone is not sufficient to determine the need for surgery. The US and FNAB are complementary to each other and should be useful when providing informed consent before thyroid surgery. PMID- 18407757 TI - Bacteriophage isolated from feedlot cattle can reduce Escherichia coli O157:H7 populations in ruminant gastrointestinal tracts. AB - Escherichia coli O157:H7 can live undetected in the gut of food animals and be spread to humans directly and indirectly. Bacteriophages are viruses that prey on bacteria, offering a natural, nonantibiotic method to reduce pathogens from the food supply. Here we show that a cocktail of phages isolated from commercial cattle feces reduced E. coli O157:H7 populations in the gut of experimentally inoculated sheep. A cocktail of phages was used in order to prevent the development of resistance to the phages. In our first in vivo study we found that our cocktail of phages reduced E. coli O157:H7 populations in the feces of sheep (p < 0.05) by 24 hours after phage treatment. Upon necropsy, populations of inoculated E. coli O157:H7 were reduced by phage treatment in both the cecum (p < 0.05) and rectum (p < 0.1). In our second in vivo study, several ratios of phage plaque-forming units (PFU) to E. coli O157:H7 colony-forming units (CFU) were used (0:1, 1:1, 10:1, and 100:1 PFU/CFU) to determine the most efficacious phage dose. A 1:1 ratio of phage to bacteria was found to be more effective (p < 0.05) than either of the higher ratios used (10:1 or 100:1). Ruminal levels of E. coli O157:H7 were not significantly reduced (p > 0.10) in any of the studies due to relatively low inoculated E. coli O157:H7 ruminal populations. Our results demonstrate that phage can be used as a preharvest intervention as part of an integrated pathogen reduction scheme. PMID- 18407758 TI - Seroprevalence of Trichinella, Toxoplasma, and Salmonella in antimicrobial-free and conventional swine production systems. AB - There has been a growing niche-market, outdoor, antimicrobial-free (ABF) swine production system in the last few years prompted by consumers' demand for a more "natural" pork product. The impact of such production systems on reemergence of current and historically significant swine-associated pathogens has not been determined. The objectives of the current study were to determine and compare Salmonella, Toxoplasma, and Trichinella seropositivity in two swine production systems: outdoor ABF and intensive indoor production systems. These three foodborne pathogens represent those with the highest importance for pork consumption. A total of 675 serum samples from three participating states, Wisconsin, North Carolina, and Ohio, were investigated. We found significantly higher seroprevalence of Salmonella and Toxoplasma from ABF herds (54% and 7%, respectively) than conventional (39% and 1%, respectively) (p = 0.001). Two pigs, both from ABF herds, were found to be seropositive for Trichinella. The results from this preliminary study suggest that all three pathogens were more commonly present in pigs that were reared in an ABF, outdoor, niche-market type of environment than the conventional, indoor-reared herds though there were some geographical variation in Salmonella. This warrants a robust epidemiologic study to determine the role of various risk factors in the two production systems that may lead to persistence of bacterial (Salmonella) pathogens and reemergence of parasites (such as Trichinella) of historical significance. PMID- 18407759 TI - Evaluation of gene expression in a single antibiotic exposure-derived isolate of Salmonella enterica typhimurium 14028 possessing resistance to multiple antibiotics. AB - Antibiotics are important tools used to control infections. Unfortunately, microbes can become resistant to antibiotics, which limit the drugs' usefulness for clinical and veterinary use. It is necessary to improve our understanding of mechanisms that contribute to or enhance antibiotic resistance. Using nalidixic acid (NA) exposure as a sole selective agent, a resistant strain of Salmonella enterica Typhimurium 14028 was derived (2a) that had acquired resistance to chloramphenicol, sulfisoxazole, cefoxitin, tetracycline, and NA. We employed gene array analysis to further characterize this derivative. Results indicate a significant difference (FDR < 5%) in the expression of 338 genes (fold regulation > 1.3) between the derivative and the parent strain growing exponentially under the same conditions at 37 degrees C. Strain 2a showed comparative induction of Salmonella pathogenicity island 2 (SPI2) transcripts and repression of SPI1 genes. Differences in expression were related to efflux pumps (increased expression), porins (decreased expression), type III secretion systems (increased expression), lipopolysaccharide synthesis (decreased expression), motility related genes (decreased expression), and PhoP/PhoQ and peptidoglycan synthesis (increased expression). It appears that 2a developed altered regulation of gene expression to decrease the influx and increase the efflux of deleterious environmental agents (antibiotics) into and out of the cell, respectively. Mechanism(s) by which this was accomplished or the reason for alterations in gene expression of other genetic systems (curli, flagella, PhoP/PhoQ, and peptidoglycan) are not immediately apparent. Evaluation of transcriptomes within multiple antibiotic-resistant mutants hopefully will enable us to better understand those generalized mechanisms by which bacteria become resistant to multiple antibiotics. Future work in sequencing these genomes and evaluating pathogenicity are suggested. PMID- 18407760 TI - Therapeutic restoration of cytolytic and suicidal cell molecular machineries favours disease healing. AB - BACKGROUND/OBJECTIVE: Several drugs have recently been demonstrated to successfully treat diseases by activating cytolytic or suicidal cell molecular machineries. On the other hand, in healthy subjects cytolytic and suicidal machineries are able to maintain tissue homeostasis, thus preventing the onset of a number of diseases. These machineries include both the cytolysis, exerted by cytotoxic T lymphocytes, of cells 'altered' by infectious or neoplastic antigens (altered cell cytolysis [AlCC]), and the suicide of antigen-activated T cells when in antigen-excess (activated cell suicide [AcCS]). RESULTS/CONCLUSIONS: These drugs may therefore, in our opinion, restore AlCC or AcCS, and, as aconsequence, favour disease healing. Thus, such a restoration could prove helpful for treating disorders caused by failure of such machineries. PMID- 18407761 TI - More trouble ahead; is gene therapy coming of age? AB - BACKGROUND: A recent incident, the halting of a Phase I/II trial utilising an adeno-associated vector, highlights the fact that there are more hurdles to overcome prior to a full realisation of gene therapy in the clinical arena. METHODS: The sources of information used to prepare the paper were obtained through published work on Pubmed/Medline and materials published on the US/UK governmental agency websites. RESULTS/CONCLUSION: Over the years, two fatal incidents associated with viral vector usage have been reported. Viral vectors used as carriers for gene therapy have failed in safety trials on two occasions. Also, the human immune response and the oncogenic property of the vectors have restricted the advancement of gene therapy as a therapeutic tool. Nonetheless, gene therapy has now progressed from its infancy 'proof of concept' stage, to the next stage in which we try to overcome the problems associated with therapeutic application. However, this progression has been slow as more and more setbacks have occurred. This calls for a new perspective and radical thinking about future vector development. PMID- 18407762 TI - Anti-inflammatory effects of mesenchymal stem cells: novel concept for future therapies. AB - BACKGROUND: Mesenchymal stem cells (MSC) are multipotent cells that can be isolated from the bone marrow and expanded in culture relatively easily. Culture expanded MSC have been used in clinical settings to enhance hematopoietic stem cell engraftment in bone marrow transplant patients and in tissue regeneration therapy. More recently, the anti-inflammatory effects of MSC have generated a great deal of interest. OBJECTIVE/METHODS: In this review we describe in vitro assays that have demonstrated how MSC regulate immune cell proliferation, differentiation and phenotype. We also highlight effector molecules produced by MSC that drive this function. In addition, we focus on animal models of lung injury, in which administration of MSC attenuates inflammation, and injury revealing a central role for MSC in mitigating pro-inflammatory networks and amplifying anti-inflammatory signals. CONCLUSIONS: The discoveries described herein have contributed to the novel concept of MSC as a therapeutic modality in inflammatory diseases, including acute lung injury. PMID- 18407763 TI - Immunological reconstitution after hematopoietic cell transplantation - its relation to the contents of the graft. AB - BACKGROUND: After hematopoietic cell transplantation (HCT), there is a prolonged period of immune deficiency. OBJECTIVE: To review the recovery of immunity after HCT, including how it is influenced by the cellular contents of the graft, and therapeutic implications. METHODS: Literature review using Medline. RESULTS/CONCLUSION: Whereas innate immunity (e.g., epithelial barriers, phagocytes, natural killer cells) typically recovers within weeks post transplant, the recovery of adaptive immunity takes longer, months for B cells and years for T cells. The recovery is influenced by the graft contents. For example the number of CD34 cells given with the graft affects the tempo of neutrophil recovery, and the number of CD4 T cells given with the graft affects CD4 T cell counts post-transplant. Improved understanding of the relationship between graft contents and immune reconstitution could lead to graft engineering improving transplant outcomes. PMID- 18407764 TI - The role of cis-regulatory elements in the design of gene therapy vectors for inherited blindness. AB - BACKGROUND: Hereditary retinal disease is currently known to involve nearly 200 different genetic loci. There has been remarkable recent progress in the treatment of retinal disease via gene therapy in animal models using virus-based vectors. The majority of retinal diseases affect one of several cell types. In order to target expression of a rescue transgene specifically to the cells in need of therapy, it is necessary to employ a cis-regulatory element (CRE) to drive expression of the transgene specifically in those cells. OBJECTIVE/METHODS: This review discusses the repertoire of CREs currently available for use in gene therapy vectors for treatment of retinal disease and outlines the issues that must be taken into consideration in the development of novel CREs for the purpose of gene therapy in the retina. CONCLUSION: There have been a number of important recent advances in the identification and characterization of retinal CREs and their utilization in gene therapy vectors. Nevertheless, future efforts to rationally manipulate existing CREs and design novel synthetic CREs for therapeutic purposes will require a better understanding of the cis-regulatory rules that govern CRE activity in vivo. PMID- 18407766 TI - Developing injectable immunoglobulins to treat cognitive impairment in Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease is a devastating disorder, clinically characterized by a comprehensive cognitive decline. The novel strategy of anti amyloid-beta immunotherapy has been suggested following encouraging results obtained in murine models of Alzheimer's disease, in non-human primates, and in small-scale clinical trials. OBJECTIVE: To examine the choice between active or passive anti-amyloid-beta immunization and the choice of the molecule to which the immune machinery should be targeted, which are central issues in future immune therapy of Alzheimer's disease. METHODS: Research into the new area of Alzheimer's disease immune therapy is primarily based on in vivo and in vitro studies of murine models of Alzheimer's disease. The studies are hence limited to defined genetic deficiencies. RESULTS/CONCLUSIONS: In humans, infusion of anti amyloid-beta antibodies is considered a safer approach than active anti-amyloid beta vaccination. Alzheimer's-disease-protective anti-amyloid-beta monoclonal antibodies should target specific epitopes within the amyloid beta(1 42) peptide, avoiding possibly harmful binding to the ubiquitous normal amyloid precursor protein. Since Alzheimer's disease immunotherapy requires repeated infusion of antibodies over a prolonged period of time, Alzheimer's disease patients will tolerate such antibodies provided the latter are exclusively of human origin. Human monoclonal antibodies that correspond to ubiquitous anti-amyloid-beta, present in all healthy humans, might bear important protective characteristics. PMID- 18407767 TI - Immunotherapy of chronic inflammatory demyelinating polyradiculoneuropathy. AB - BACKGROUND: chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an immune-mediated disease. Current treatments are aimed at modulating the immune response to achieve remission and maintain functional status. However, many patients fail to make a long-term recovery with current treatments. OBJECTIVES: to review the literature on immunotherapy for CIDP. METHODS: We used the Search Strategy of the Cochrane Neuromuscular Disease Review Group to search Medline and Embase. Randomised and non-randomised studies examining the effects of any therapeutic agent in patients with CIDP were selected. The references of relevant articles were scanned to identify additional reports of interest. RESULTS/CONCLUSION: An overview of the different treatments for CIDP is provided. Emphasis has been placed on evidence from randomised controlled trials but open non-randomised studies are discussed if appropriate. We include a treatment algorithm and provide our views on current treatments, ongoing trials and possible directions for further research. PMID- 18407768 TI - Advancing age and immune cell dysfunction: is it reversible or not? AB - BACKGROUND: Decline in immune function with advancing age is a hallmark of aging. As a result, infectious diseases cause more morbidity and mortality to the elderly. These age-related changes significantly alter the functionality of both innate and adaptive immune components. OBJECTIVES: Although age-associated dysfunction of the immune system is well reported, not much is known about the extrinsic and intrinsic factors that progressively bring about functional changes in immune cell types. METHODS: This review focuses on our current understanding of age-associated changes in innate and adaptive immune cell types and how this knowledge could be used to improve the immune competence of elderly individuals. RESULTS/CONCLUSION: Therapeutic approaches to rejuvenate the exhausted immune system in the elderly need more attention. PMID- 18407769 TI - The recombinant humanized anti-IL-6 receptor antibody tocilizumab, an innovative drug for the treatment of rheumatoid arthritis. AB - BACKGROUND: IL-6 is a pro-inflammatory cytokine with multiple roles in the pathogenesis of rheumatoid arthritis (RA). Targeting IL-6 with the humanized anti IL-6 receptor antibody tocilizumab was effective in several placebo-controlled clinical studies in RA. OBJECTIVES: To address how clinically efficacious blockade of IL-6 signalling with inteleukin-6 receptor antibody is in RA patients and what the potential mode of action explaining tocilizumab activity in RA treatment could be. RESULTS/CONCLUSION: IL-6 induces autoantibody-producing plasma cells and effector T cells and is implicated in the development of clinical signs and symptoms, including increased synthesis of acute phase reactants, fatigue, anaemia and anorexia. Its effects also included significant improvements in American College of Rheumatology (ACR)20, ACR50 and ACR70 values, as well as in health-related quality of life measures, compared with controls. Tocilizumab also prevents radiographic progression of joint damage. Tocilizumab is generally well tolerated and efficacious in patients refractive to conventional DMARD therapies. PMID- 18407765 TI - Antibody-cytokine fusion proteins: applications in cancer therapy. AB - BACKGROUND: Antibody-cytokine fusion proteins consist of cytokines fused to an antibody to improve antibody-targeted cancer immunotherapy. These molecules have the capacity to enhance the tumoricidal activity of the antibodies and/or activate a secondary antitumor immune response. OBJECTIVE: To review the strategies used to develop antibody-cytokine fusion proteins and their in vitro and in vivo properties, including preclinical and clinical studies focusing on IL 2, IL-12 and GM-CSF. METHODS: Articles were found by searching databases such as PubMed and Clinical Trials of the US National Institutes of Health. RESULTS/CONCLUSION: Multiple antibody-cytokine fusion proteins have demonstrated significant antitumor activity as direct therapeutics or as adjuvants of cancer vaccines in preclinical studies, paving the way for their clinical evaluation. PMID- 18407770 TI - Alfimeprase for the treatment of acute peripheral arterial occlusion. AB - BACKGROUND: Catheter-directed thrombolysis for the management of acute peripheral arterial occlusion emerged as a viable treatment option in the 1990s. It offers a less invasive approach than traditional open surgery for correcting acute limb ischemia. Nonetheless, thrombolysis is plagued by a relatively high rate of bleeding complications as well as long infusion times. OBJECTIVE: To review the clinical experience with alfimeprase, a new thrombolytic agent. METHODS: All published data on alfimeprase were reviewed. Review articles, press releases and web-based data were also included. RESULTS/CONCLUSIONS: Alfimeprase is a novel agent with a unique mechanism of action compared with currently available thrombolytic agents. It is a direct-acting fibrinolytic agent that does not require activation of plasminogen. This mechanism may potentially reduce the number of bleeding complications. Current clinical data are limited, but ongoing clinical trials may demonstrate that this compelling agent represents a clinical advance. PMID- 18407771 TI - The role of pegvisomant in the treatment of acromegaly. AB - BACKGROUND: Expert surgery in acromegaly is curative in about only 60% of cases. Postsurgical radiation therapy is associated with a high incidence of hypopituitarism and its effect on growth hormone decrease is slow. Therefore, effective adjuvant medical treatment is mandatory. OBJECTIVE: To delineate the role of pegvisomant in the treatment of acromegaly. METHODS: A search of the Medline, Embase and Web of Science databases for clinical studies on pegvisomant was performed. In addition, abstracts of the major endocrine society meetings in 2007 were searched. RESULTS: When treatment with somatostatin analogues after non curative surgery does not lead to normalisation of serum IGF-I and growth hormone concentrations, pegvisomant alone or in combination with somatostatin analogues can control disease in a substantial number of patients. The most important side effects are disturbed liver function tests and, rarely, growth of the adenoma. The efficacy in non-operated patients and the long-term safety have yet to be ascertained. PMID- 18407772 TI - Heavy element enhanced synchrotron stereotactic radiotherapy as a promising brain tumour treatment. AB - Synchrotron stereotactic radiotherapy (SSR) is a treatment that involves selective accumulation of high-Z elements in tumours followed by stereotactic irradiation, in CT mode, with monochromatic X-rays from a synchrotron source, tuned at an optimal energy. The irradiation geometry, characteristic X-rays, photoelectrons, and Auger electrons generated on high-Z atoms by kilovoltage X rays produce a localized dose enhancement. Two complimentary SSR approaches have been successfully developed in the past 5 years in our team, and may be promising in high-grade glioma management: iodine-enhanced SSR, with an iodinated contrast agent; and Pt-enhanced SSR; a concomitant radio-chemotherapy treatment with locoregional injection of platinated chemotherapy drugs. The results for iodine enhanced SSR using contrast agents are presented in this paper. IUdR-enhanced SSR was also tested in this study. Up to 15 Gy, intracarotid infusion of iodine significantly improved the rats' survival compared to irradiation alone. SSR provides the most protracted survivals of F98 glioma-bearing rats. The technique is currently transferred to clinical trials. Iodine-enhanced SSR will be implemented first, because of its simplicity; and pave the way for Pt-enhanced SSR, the most efficient technique, but still needing to be improved in terms of intrinsic toxicity. PMID- 18407773 TI - The use of Fresh Frozen Plasma (FFP) in 2007 in France. AB - After a more than four-fold decrease in Fresh Frozen Plasma (FFP) consumption in the nineties in France, probably due to the consequences of HIV epidemics, the use of FFP is again increasing in recent years but at a slower rate. In the surgical and trauma area, recent data suggest that guidelines for the use of FFP may need to be modified. Indeed, contrary to traditional beliefs and guidelines, several studies evaluating conditions with severe hemorrhage (very often associated with coagulation abnormalities) have shown that early use of FFP may be associated with better patient outcome. This has indeed been shown in emergency major vascular surgery and in trauma patients. Although there is a trend to favor a larger use of FFP in specific circumstances (i.e., major hemorrhage), reasons to better control administration of FFP remain. Several audits have indeed shown that the rate of inappropriate FFP transfusion remains high, from 20 to almost 100% of cases. Moreover, FFP continues to be used in patients who may be better treated with other strategies. The best example is the frequently inappropriate use of FFP in bleeding patients with excess anticoagulation from vitamin K antagonists. Even recent studies have shown that prothrombin complex administration is efficient, safe and provides very rapid reversal. Many physicians continue however to administer FFP as a first line treatment with notably increasing the risk of fluid overload, delayed efficacy and increased risk of transfusion-related acute lung injury (TRALI). PMID- 18407774 TI - Use of plasma: clinical indications and types of plasma components in Sweden. AB - The use of plasma in Sweden is relatively high compared to other countries in the European Union. An analysis of all transfusion recipients in Orebro county during the whole year 2000 was performed. There were 3159 transfusion recipients of whom 96% had a registered diagnosis and 50% had undergone a "true" operation. Seven hundred and eleven patients (23%) had received plasma. Significantly more operated than nonoperated and more men than women received plasma. The typical plasma recipient was a man undergoing cardiovascular surgery. In Sweden there are two main types of plasma components: fresh frozen (FFP) and nonfrozen liquid plasma stored for up to 14 days, both considered to be clinically equal for most indications. The quality of these components as well as stored thawed FFP has been studied. The major storage effect was cold-induced contact activation and thereby consumption of C1 esterase inhibitor (C1INH) by day 14 in 22%. The citrate content in plasma sustained the overall coagulation function over 14 days. Other studies have shown that the levels of FV and ADAMTS 13 after 14 days remain at 70% or more compared to those for FFP. Since it is immediately available, liquid, nonfrozen or thawed, plasma is of great value in emergencies. Quality criteria for plasma components need to be assessed against evidence based indications and published in guidelines. PMID- 18407775 TI - HIV evolution in response to HLA-restricted CTL selection pressures: a population based perspective. AB - Cytotoxic T lymphocytes (CTL) recognize antigenic peptides displayed by HLA class I molecules on the infected cell surface and represent a major selective force driving HIV evolution through a phenomenon known as "immune escape". Here we summarize recent advances in our understanding of the consequences of CTL escape on HIV evolution at the population level and discuss its implications for HIV vaccine design. PMID- 18407776 TI - A rapid recombination assay of HIV-1 using murine CD52 as a novel biomarker. AB - Biomarkers are commonly used for verification of infection in conjunction with the development of viral vectors or experiments involving virus infection. Leukocyte surface antigens (CDs) are a prime option for biomarkers since they can be easily visualized and analyzed by flow cytometry after indirect fluorescent staining. For analyses of human cells, murine CD24 (Heat Stable Antigen: HSA) and CD90.2 (Thy-1.2) are currently being used. In the study reported here, we attempted to develop a rapid system for measuring retroviral genome recombination efficiency. For this purpose, we looked for an alternative CD molecule which could be used as a marker on a viral vector concurrently with other markers. We found that murine CD52 is suitable for this purpose because of its small gene size, low inhibitory effect on virus production, and measurable level of surface expression. With this novel biomarker, we succeeded in developing a rapid viral recombination measuring system using a flow cytometer. PMID- 18407777 TI - Clinical evaluation of two bilateral hand allotransplantations at six and three years follow-up. AB - We report a six and three years follow-up clinical evaluation of two bilateral hand allotransplantations from brain-dead multi-organs donors performed in two young adult traumatic hand amputees. Lifelong immunosuppressive treatment included tacrolimus, mycophenolate mofetil and prednisolone. In both patients, early complications were observed but were successfully treated. At follow-up, clinical results allow useful hand function in both patients, by far superior to what can be provided by current myoelectric prostheses. Although the long-term risks-benefits ratio of bilateral hand transplantation is still unknown, these two cases demonstrate that this new treatment is feasible. Further experimental and clinical research is needed to better delineate its role in the future of hand surgery. PMID- 18407778 TI - [What kind of follow-up after surgical treatment for high-grade cervix lesion?]. AB - The women treated for a high-grade cervical lesion (CIN2+) have a high and prolonged (beyond 25 years) risk of recurrence (x 5 compared to the general population) of cervical as well as extracervical lesion. The cytological and colposcopic protocols of follow-up raise the problem of their insufficient sensitivity, and the compliance of the patients to this prolonged follow-up is low. Test HPV seeking the presence of high-risk HPV benefits from a high sensitivity and presents altogether a very strong negative predictive value. The addition of this test to the cytological monitoring three to six months after the conization makes it possible to distinguish a group of patients with low risk (with both tests negative) being able to profit from a traditional follow-up, from a high-risk group having at least one positive test, whose surveillance must be reinforced (with triage by colposcopy), prolonged in time and extended beyond the cervix. Like as the primary screening of cervical lesions, the follow-up of the patients after conization must profit from the addition of test HPV and would deserve protocolization and organization. PMID- 18407779 TI - Topology optimization of a novel stent platform with drug reservoirs. AB - The new generation of drug-eluting stents (DES) is required to control drug release kinetics. A novel DES (the Conor stent) with drug reservoirs on struts has been engineered. Topology optimization of one Conor stent strut was based on the commercial finite element analysis code OptiStruct, with the aim of increasing the strut stiffness while retaining its drug holding capacity. Results show that the element density distribution of the strut model was optimized with manufacturing constraints of extrusion constraint and minimum member size control. The optimal result was directly transformed to a clear, manufacturable design concept using the OptiStruct utility OSSmooth. The final manufacturing design increased the strut stiffness and yielded better stress distribution, as compared to the original strut design under the same loading. Topology optimization may help designers devise novel stent platforms for future DES with drug reservoirs and adequate scaffolding. PMID- 18407780 TI - Characterization of the molecular interaction between caveolin-1 and the P2X receptors 4 and 7 in E10 mouse lung alveolar epithelial cells. AB - P2X(4) and P2X(7) receptors are abundantly expressed in alveolar epithelial cells, and are thought to play a role in regulating fluid haemostasis. Here, we analyzed the expression and localization of the P2X(4)R, and characterized the interaction between Cav-1 and both P2X(4)R and P2X(7)R in the mouse alveolar epithelial cell line E10. Using the biotinylation assay, we found that only glycosylated P2X(4)R is exposed at the cell surface. Triton X-100 solubility experiments and sucrose gradient centrifugation revealed that P2X(4)R was partially localized in Cav-1 rich membrane fractions. Cholesterol depletion with Mbeta-CD displaced Cav-1 and P2X(4)R from the low-density to the high-density fractions. Suppression of Cav-1 protein expression using short hairpin RNAs resulted in a large reduction in P2X(4)R levels. Double immunofluorescence showed that P2X(4)R and Cav-1 partially colocalize in vitro. Using the GST pull-down assay, we showed that Cav-1 interacts in vitro with both P2X(4)R and P2X(7)R. Co immunoprecipitation experiments confirmed the interaction between P2X(7)R and Cav 1. ATP stimulation increased the level of P2X(4)R in the lipid raft/caveolae fraction, whereas Cav-1 content remained constant. Our results support recent evidence that P2X receptors are present in both raft and non-raft compartments of the plasma membrane and thus exhibit variable ATP sensitivity. PMID- 18407781 TI - Molecular regulation of cardiac hypertrophy. AB - Heart failure is one of the leading causes of mortality in the western world and encompasses a wide spectrum of cardiac pathologies. When the heart experiences extended periods of elevated workload, it undergoes hypertrophic enlargement in response to the increased demand. Cardiovascular disease, such as that caused by myocardial infarction, obesity or drug abuse promotes cardiac myocyte hypertrophy and subsequent heart failure. A number of signalling modulators in the vasculature milieu are known to regulate heart mass including those that influence gene expression, apoptosis, cytokine release and growth factor signalling. Recent evidence using genetic and cellular models of cardiac hypertrophy suggests that pathological hypertrophy can be prevented or reversed and has promoted an enormous drive in drug discovery research aiming to identify novel and specific regulators of hypertrophy. In this review we describe the molecular characteristics of cardiac hypertrophy such as the aberrant re expression of the fetal gene program. We discuss the various molecular pathways responsible for the co-ordinated control of the hypertrophic program including: natriuretic peptides, the adrenergic system, adhesion and cytoskeletal proteins, IL-6 cytokine family, MEK-ERK1/2 signalling, histone acetylation, calcium mediated modulation and the exciting recent discovery of the role of microRNAs in controlling cardiac hypertrophy. Characterisation of the signalling pathways leading to cardiac hypertrophy has led to a wealth of knowledge about this condition both physiological and pathological. The challenge will be translating this knowledge into potential pharmacological therapies for the treatment of cardiac pathologies. PMID- 18407782 TI - Proteases and antiproteases in development, homeostasis and disease: The old, the new, and the unknown... PMID- 18407783 TI - Molecular crowding and order in photosynthetic membranes. AB - The integrity and maintenance of the photosynthetic apparatus in thylakoid membranes of higher plants requires lateral mobility of their components between stacked grana thylakoids and unstacked stroma lamellae. Computer simulations based on realistic protein densities suggest serious problems for lateral protein and plastoquinone diffusion especially in grana membranes, owing to strong retardation by protein complexes. It has been suggested that three structural features of grana thylakoids ensure efficient lateral transport: the organization of protein complexes into supercomplexes; the arrangement of supercomplexes into structured assemblies, which facilitates diffusion process in crowded membranes; the limitation of the diameter of grana discs to less than approximately 500 nm, which keeps diffusion times short enough to support regulation of light harvesting and repair of photodamaged photosystem II. PMID- 18407784 TI - Using Arabidopsis to explore zinc tolerance and hyperaccumulation. AB - Identifying the particular gene or genes underlying a specific adaptation is a major challenge in modern biology. Currently, the study of naturally occurring variation in Arabidopsis thaliana provides a bridge between functional genetics and evolutionary analyses. Nevertheless, the use of A. thaliana to study adaptation is limited to those traits that have undergone selection. Therefore, to understand fully the genetics of adaptation, the vast arsenal of genetic resources developed in A. thaliana must be extended to other species that display traits absent in this model species. Here, we discuss how A. thaliana resources can significantly enhance the study of heavy-metal tolerance and hyperaccumulation in the wild species Arabidopsis halleri. PMID- 18407785 TI - The use of high modulus silicone hydrogel (SiHy) lens in the management of epithelial defects. AB - Corneal dystrophies are relatively rare bilateral anomalies. Meesman dystrophy is typically asymptomatic with little effect on visual performance. This case highlights the use of a high modulus silicone hydrogel (SiHy) lens in the management of an atypical presentation of Meesman's dystrophy with associated visual impairment due to epithelial defects. The selection of a SiHy material provided increased oxygenation to re-establish corneal integrity. However, selecting the appropriate modulus was an additional factor to consider in this case since it had a direct effect on the visual outcome. The higher modulus SiHy lenses provided a better visual success for this patient than one with a lower modulus. Modulus consideration may prove to be an additional factor in the lens choice of SiHy in the management of epithelial defects. PMID- 18407786 TI - Epigenetic interplay between histone modifications and DNA methylation in gene silencing. AB - Knowledge on heritable changes in gene expression that result from epigenetic events is of increasing relevance in the development of strategies for prevention, early diagnosis and treatment of cancer. Histone acetylation and DNA methylation are epigenetic modifications whose patterns can be regarded as heritable marks that ensure accurate transmission of the chromatin states and gene expression profiles over many cell generations. Importantly, patterns and levels of DNA methylation and histone acetylation are profoundly altered in human cancers. Accumulating evidence suggests that an epigenetic cross-talk, i.e. interplay between DNA methylation and histone acetylation, may be involved in the process of gene transcription and aberrant gene silencing in tumours. Although the molecular mechanism of gene activation is relatively well understood, the hierarchical order of events and dependencies leading to gene silencing in the course of cancer development remain largely unknown. While some studies suggest that DNA methylation patterns guide histone modifications (including histone acetylation and methylation) during gene silencing, other studies argue that DNA methylation takes its cues primarily from histone modification states. In this review, we summarize current knowledge on the interplay between DNA methylation and histone modifications during gene silencing and its importance in the integration of environmental and intrinsic stimuli in the control of gene expression. We also discuss the importance of an epigenetic cross-talk in the protection against genetic changes in response to environmental genotoxins as well as the implication for cancer therapy and prevention. PMID- 18407787 TI - Study of inversion isomerism in some 4,5-cis-substituted-2-isoxazolidineethanols by NMR spectroscopy. AB - A series of beta-phenyl-2-isoxazolidineethanols having cis-disposed substituents at C(4) and C(5) has been prepared by asymmetric nitrone cycloaddition reactions and their NMR spectra recorded over a wide range of temperatures. The spectra in CDCl3 at low temperatures indicate the presence of two interconverting invertomers for some of the tri-substituted isoxazolidines. The effect of H bonding--intramolecular in CDCl3 and intermolecular in CD3OD--on the population ratio of the nitrogen invertomers has been investigated. The nitrogen inversion barriers are determined using complete line-shape analysis, and their dependence on solvent is discussed. PMID- 18407789 TI - Transcatheter closure of secundum atrial septal defects with the Amplatzer septal occluder in adults and children-follow-up closure rates, degree of mitral regurgitation and evolution of arrhythmias. AB - AIM: To report the results of transcatheter atrial septal defect (ASD) closure with the Amplatzer septal occluder (ASO) from the single centre providing interventions for congenital heart disease in New Zealand. METHODS: A single centre retrospective review of all patients 1997-2004 inclusive, undergoing planned transcatheter ASD closure was undertaken. Implantation success, complications and latest patient follow-up are described. RESULTS: Percutaneous ASD closure was planned in 227 adults and children. (55% had additional medical co-morbidities.) The mean ASD size was 22+/-6 mm, and device size ranged from 5 to 40 mm. Closure was successful in 93%, unsuccessful in 5% and not attempted in 2%. There were two device embolisations, one immediately post-release and one within 24h. Minor procedural complications occurred in 5% of patients. There were no cases of suspected or proven device erosion. Ninety-two percent had documented follow-up beyond 6 months. At latest follow-up the closure rate was 98.5%. There was no significant change in the degree of mitral regurgitation. Sixteen of 24 with pre-closure arrhythmias resolved while 10 remained in atrial fibrillation. Six patients developed new arrhythmias but these were well controlled medically. CONCLUSION: There is a high implantation and closure rate using transcatheter ASD closure with the Amplatzer septal occluder in children and adults avoiding the need for cardiac surgery. Follow-up at one year shows there is no progression of mitral regurgitation, and supraventricular arrhythmias have usually resolved. PMID- 18407790 TI - Natural history of cervical neoplasia and risk of invasive cancer in women with cervical intraepithelial neoplasia 3: a retrospective cohort study. AB - BACKGROUND: The invasive potential of cervical intraepithelial neoplasia 3 (CIN3; also termed stage 0 carcinoma) has been poorly defined. At the National Women's Hospital, Auckland, New Zealand, treatment of CIN3 was withheld from a substantial number of women between 1965 and 1974 as part of an unethical clinical study. The resulting variation in management allows comparison of the long-term risk of invasive cancer of the cervix in women whose lesion was minimally disturbed with those who had adequate initial treatment followed by conventional management. We aimed to estimate the long-term risk of invasive cancer in these two groups of women. A judicial inquiry referred for independent clinical review in 1988 all women for whom there remained doubt about the adequacy of their management. METHODS: Between February, 2001, and December, 2004, medical records, cytology, and histopathology were reviewed for all women with CIN3 diagnosed between 1955 and 1976, whose treatment was reviewed by judicial inquiry and whose medical records could be located, and linkages were done with cancer and death registers and electoral rolls. To take into account the probability that the CIN3 lesion had been completely removed, we classified adequacy of treatment by type of procedure, presence of CIN3 at the excision margin, and subsequent cytology. The primary outcome was cumulative incidence of invasive cancer of the cervix or vaginal vault. Follow-up continued until death or Dec 31, 2000, whichever came first. Analyses accounted for procedures during follow-up. FINDINGS: 1229 women whose treatment was reviewed by the judicial inquiry in 1987-88 were included. Of these, 48 records (4%) could not be located and 47 women (4%) did not meet the inclusion criteria. At histopathological review, a further 71 (6% of 1134) women were excluded because the review diagnosis was not CIN3. We identified outcomes in the remaining 1063 (86% of 1229) women diagnosed with CIN3 at the hospital in 1955-76. In 143 women managed only by punch or wedge biopsy, cumulative incidence of invasive cancer of the cervix or vaginal vault was 31.3% (95% CI 22.7-42.3) at 30 years, and 50.3% (37.3 64.9) in the subset of 92 such women who had persistent disease within 24 months. However, cancer risk at 30 years was only 0.7% (0.3-1.9) in 593 women whose initial treatment was deemed adequate or probably adequate, and whose treatment for recurrent disease was conventional. INTERPRETATION: This study provides the most valid direct estimates yet available of the rate of progression from CIN3 to invasive cancer. Women with untreated CIN3 are at high risk of cervical cancer, whereas the risk is very low in women treated conventionally throughout. PMID- 18407791 TI - On the relationship between GC content and the number of predicted microRNA binding sites by MicroInspector. AB - MicroRNA GC content and length is believed to play a role in the prediction of putative microRNA targets. MicroInspector was evaluated to determine the extent to which these characteristics of microRNAs play a role in binding site predictive accuracy. A strong bias towards under predicting the number of expected bindings sites for low GC content sequences was observed, especially for microRNAs with <50% GC content. Researchers working with organisms with unusually low GC content should be aware of this bias. PMID- 18407792 TI - Arachidonic acid-induced expression of the organic solute and steroid transporter beta (Ost-beta) in a cartilaginous fish cell line. AB - The organic solute and steroid transporter (OST/Ost) is a unique membrane transport protein heterodimer composed of subunits designated alpha and beta, that transports conjugated steroids and prostaglandin E(2) across the plasma membrane. Ost was first identified in the liver of the cartilaginous fish Leucoraja erinacea, the little skate, and subsequently was found in many other species, including humans and rodents. The present study describes the isolation of a new cell line, LEE-1, derived from an early embryo of L. erinacea, and characterizes the expression of Ost in these cells. The mRNA size and amino acid sequence of Ost-beta in LEE-1 were identical to that previously reported for Ost beta from skate liver, and the primary structure was identical to that of the spiny dogfish shark (Squalus acanthias) with the exception of a single amino acid. Ost-beta was found both on the plasma membrane and intracellularly in LEE-1 cells, consistent with its localization in other cell types. Interestingly, arachidonic acid, the precursor to eicosanoids, strongly induced Ost-beta expression in LEE-1 cells and a lipid mixture containing arachidonic acid also induced Ost-alpha. Overall, the present study describes the isolation of a novel marine cell line, and shows that this cell line expresses relatively high levels of Ost when cultured in the presence of arachidonic acid. Although the function of this transport protein in embryo-derived cells is unknown, it may play a role in the disposition of eicosanoids or steroid-derived molecules. PMID- 18407794 TI - An intrapancreatic cholangiocarcinoma detected with optical coherence tomography during endoscopic retrograde cholangiopancreatography. PMID- 18407793 TI - Anti-angiogenic potential of CoenzymeQ10, riboflavin and niacin in breast cancer patients undergoing tamoxifen therapy. AB - Tumour angiogenesis is a complex mechanism consisting of multi-step events including secretion or activation of angiogenic factors by tumour cells, activation of proteolytic enzymes, proliferation, migration and differentiation of endothelial cells. Both primary and metastatic tumours in the breast are dependent on angiogenesis. In the present study, 84 breast cancer patients were randomized to receive a daily supplement of CoQ(10) 100 mg, riboflavin 10 mg and niacin 50 mg (CoRN), one dosage per day along with tamoxifen (TAM) 10 mg twice a day. Serum pro-angiogenic levels were elevated in untreated breast cancer patients (Group II) and their levels were found to be reduced in breast cancer patients undergoing TAM therapy for more than 1 year (Group III). When these group III breast cancer patients were supplemented with CoRN for 45 days (Group IV) and 90 days (Group V) along with TAM, a further significant reduction in pro angiogenic marker levels were observed. Supplementing CoRN to breast cancer patients has found to decrease the levels of pro-angiogenic factors and increase the levels of anti-angiogenic factors. A reduction in pro-angiogenic marker levels attributes to reduction in tumour burden and may suggest good prognosis and efficacy of the treatment, and might even offer protection from cancer metastases and recurrence. PMID- 18407795 TI - Exudative hemorrhagic duodenitis as a primary event in a child with Henoch Schonlein purpura. PMID- 18407796 TI - A case of gastric syphilis. PMID- 18407797 TI - Hepatitis C infection with normal liver chemistry tests. PMID- 18407799 TI - Value of spatiotemporal representation of manometric data. AB - BACKGROUND & AIMS: High-resolution manometry with spatiotemporal representation of pressure data is a technique that has developed during the past 10-15 years. We compared spatiotemporal and traditional line plot representation of manometry data in a group of medical students in terms of the ability of the user to come to a rapid and accurate diagnosis and evaluated user preferences for the 2 forms of data display. METHODS: After standardized paper-based and electronic tutorials, 60 medical students classified 30 typical examples of a range of motility disorders in both line plots (10 sensors, including a "virtual sleeve") and spatiotemporal plot format (derived from 16 sensors). Swallows were presented electronically in random order. The accuracy and speed of the assessment were compared between the 2 forms of data presentation, as well as a subjective rating of preference. Results are presented as mean +/- standard error of the mean. RESULTS: Classifications based on data presented in spatiotemporal format were more often correct (89% +/- 1.2% vs 86% +/- 1.3%, P = .002), and correct diagnoses were provided more promptly (25 +/- 2.9 seconds vs 31 +/- 3.2 seconds, P < .001) than in line plot format. Sixty-eight percent of the study population preferred the spatiotemporal presentation. CONCLUSIONS: The analysis of manometry data by manometry-naive individuals is faster and more accurate when data are presented in spatiotemporal than in line plot format. In addition, users preferred the spatiotemporal plots. Spatiotemporal presentation of manometric data is likely to be more easily understood by patients and the "non-expert" physician community. PMID- 18407798 TI - Peginterferon pharmacokinetics in African American and Caucasian American patients with hepatitis C virus genotype 1 infection. AB - BACKGROUND & AIMS: The relationship between serum peginterferon pharmacokinetics and pharmacodynamics and the early virologic response (EVR) to peginterferon and ribavirin therapy was assessed in patients with chronic hepatitis C virus (HCV) genotype 1 infection. METHODS: A total of 333 patients (160 African Americans [AA] and 173 Caucasian Americans [CA]) who received peginterferon alpha-2a (180 microg/wk) without a dose modification during the initial 4 weeks of therapy were analyzed. Peginterferon and 2,5-oligoadenylate synthetase (2,5-OAS) serum levels were measured on days 0, 1, 2, 3, 7, 14, 28, 56, 84, and 168 of treatment. The EVR (>or=2-log(10) decline in HCV RNA levels by week 12 of therapy) was the primary virologic end point. RESULTS: Peginterferon pharmacokinetics after the first dose were similar in AA and CA, but AA had greater peginterferon concentrations at days 1, 3, 14, and 28 (P < .05). AA had higher absolute serum 2,5-OAS levels on days 0, 1, 2, 3, 7, 14, 28, and 56 (P < .05), but the magnitude of 2,5-OAS induction during treatment were similar. AA patients showed a smaller decline in serum HCV RNA during the first 28 days of treatment (P < .001) and a lower EVR (65% vs 83%). AA and CA with EVR had significantly higher serum peginterferon concentrations and serum 2,5-OAS induction during the first 12 weeks than patients without an EVR. CONCLUSIONS: Peginterferon alpha-2a pharmacokinetic and pharmacodynamic variability is associated with EVR in both AA and CA with HCV infection, but do not explain the racial disparity in combination treatment efficacy. PMID- 18407800 TI - Eosinophilic esophagitis: analysis of food impaction and perforation in 251 adolescent and adult patients. AB - BACKGROUND & AIMS: Eosinophilic esophagitis is a rapidly emerging, chronic inflammatory disorder. Prolonged inflammation evokes structural alterations and a fragile esophageal wall prone to perforation/rupture and food impaction. This report assesses the risk of spontaneously arising and procedure-induced complications and proposes practical recommendations. METHODS: The Swiss Esophageal Esophagitis Database documented 251 confirmed cases. A chart review identified which patients had required endoscopic bolus removal and/or experienced transmural esophageal perforation/rupture. In addition, a MEDLINE search for "eosinophilic esophagitis" with "esophageal perforation" or "esophageal rupture" was undertaken. RESULTS: During an 18-year period, 87 patients (34.7%) experienced 134 food impactions requiring flexible (124, 92.5%) or rigid (10, 7.5%) endoscopic bolus removal. Transmural perforation occurred in 20% (2/10) of rigid procedures, and 1 esophageal rupture (Boerhaave's syndrome) was observed. CONCLUSIONS: Bolus removal by rigid endoscopy is a high-risk procedure and should be avoided in eosinophilic esophagitis patients who require a gentler approach. Whether food impaction and esophageal wall remodeling can be prevented with anti-inflammatory medication is still undetermined. All Boerhaave's syndrome cases should be evaluated for underlying eosinophilic esophagitis. PMID- 18407801 TI - Laparoscopic sleeve gastrectomy with endoscopic guidance in morbid obesity. AB - BACKGROUND: Sleeve gastrectomy (SG) has been shown to be an effective first-stage procedure for morbidly obese patients. The SG is presently performed over a bougie of varying sizes, which is useful, but known to produce injuries on insertion. In a retrospective study, we evaluated the effect of the laparoscopic SG (LSG) on excess weight loss during 1 year of follow-up using a 29F endoscope instead of a bougie. METHODS: During a 1-year period, LSG was performed on 20 (18 women and 2 men) consecutive patients. Gamma regression analysis was used to determine whether the variation in gender, age, initial body mass index, Hispanic ethnicity, and interval after surgery were related to excess weight loss. RESULTS: No deaths and 1 minor complication of oozing from the staple line occurred. The excess weight loss increased steadily over time, with a median 20% at 3 months, 32% at 6 months, 42% at 9 months, and 53% at 12 months. The median initial body mass index was 44.5 kg/m2, and the median age was 50 years. Of the 20 patients, 2 were men (10%) and 18 women (90%); 5 (25%) were Hispanic and 15 (75%) were non-Hispanic. The patients had a median 11.5 co-morbidities. Nausea was common for about 7 days postoperatively. An increase in the initial body mass index and increased co-morbidities were the only 2 variables directly and statistically connected with the percentage of excess weight loss (P <.05). CONCLUSION: The results of our study have shown that LSG with endoscopic guidance appears safe and effective and could be tried using a larger set of patients as a single-stage operation. PMID- 18407803 TI - Incidence and management of enteric leaks after gastric bypass for morbid obesity during a 10-year period. AB - BACKGROUND: To describe the incidence, etiology, outcomes, and management of enteric leaks in patients who had undergone open or laparoscopic gastric bypass. METHODS: From November 1996 to November 2006, 1133 patients underwent primary gastric bypass at Columbia University, New York-Presbyterian Hospital. A retrospective review of our prospective bariatric surgery registry identified 17 patients (1.5%) who developed a clinically apparent enteric leak after surgery. The demographic and outcome data were studied. RESULTS: The mean body mass index was 52 kg/m(2) (range 35-65), and 15 (88%) of the 17 patients were women. The mean number of co-morbidities per patient was 1.3 and included hypertension in 11, diabetes in 9, and sleep apnea in 6. Ten patients had previously undergone abdominal surgery. The enteric leak was diagnosed by radiographic studies in 12 patients (9 of 12 by upper gastrointestinal series and 3 of 6 by computed tomography); the remaining 5 patients were diagnosed at re-exploration. Of the 17 patients, 12 (70%) were treated by laparoscopy and 5 (29%) by laparotomy. The mean time from completion of the index procedure to the diagnosis of the leak was 2 days (range 1-5) for patients treated by laparoscopy versus 4 days (range 1-6) for patients treated by laparotomy (P <.05). The patients treated by laparoscopy experienced a shorter hospital stay, but the difference was not statistically significant (mean 11.4 days, range 6-36, versus 18 days, range 7-33; P >.05). One of the laparoscopic patients (5.9%) died. CONCLUSION: Enteric leak is a significant complication after gastric bypass. Prompt treatment should be based on clinical suspicion, because contrast and cross-sectional imaging studies might not be reliable diagnostic tests. A laparoscopic index procedure might be associated with an earlier diagnosis. PMID- 18407804 TI - Diagnostic laparoscopy for chronic abdominal pain after gastric bypass. AB - BACKGROUND: To evaluate, in an observational study, the utility of diagnostic laparoscopy as a tool to evaluate patients with abdominal pain of unknown etiology after gastric bypass surgery. METHODS: A retrospective analysis was performed of data from patients who had undergone laparoscopy for diagnosis or treatment of abdominal pain. This study included 13 patients with negative preoperative radiographic and/or endoscopic findings. RESULTS: A total of 13 patients who had undergone Roux-en-Y gastric bypass underwent diagnostic laparoscopy for abdominal pain. The findings included internal hernia (4), adhesions (3), ventral hernia (2), partial small bowel obstruction (1), and chronic cholecystitis (1). There were 2 negative laparoscopies, while a diagnosis was made in 85%. After an average follow-up of 3.2 months, 7 of 11 patients had unresolved abdominal pain and 4 patients experienced pain resolution (2 patients were lost to follow-up). CONCLUSION: The results from this small retrospective study suggest that significant pathologic findings can be identified in most patients who have negative preoperative evaluation findings; however, the efficacy of diagnostic laparoscopy to eliminate pain in this patient population requires additional study. Despite the potential complications, we believe that diagnostic laparoscopy has a role in the diagnosis and treatment of chronic abdominal pain after gastric bypass. PMID- 18407805 TI - Intussusception after Roux-en-Y gastric bypass for morbid obesity. PMID- 18407806 TI - Cardiovascular responses to electrical stimulation of sympathetic nerves in the pithed mouse. AB - The pithed rat model has been used extensively to study peripheral cardiovascular responses to electrical stimulation of the sympathetic nervous system, as pithing eliminates central and reflex effects. However, since the transgenic mouse has become a standard and economical model organism, an electrically stimulated pithed mouse would facilitate a variety of studies. We have developed surgical techniques, drug doses and stimulation parameters for an electrically stimulated pithed mouse to study peripheral sympathetic nerve effects on blood pressure. Similar to the pithed rat, the pithed mouse showed voltage and frequency dependent blood pressure responses to a pulsed train of electrical stimuli. In addition, alpha-adrenergic stimulation with phenylephrine gave a marked systolic pressor response, while the beta2 agonist salbutamol lowered diastolic blood pressure. Furthermore, pithed transgenic mice unable to synthesize catecholamines in adrenergic cells displayed smaller pressor responses than pithed control mice. In summary, the electrically stimulated pithed mouse can be used to study peripheral effects of the sympathetic system on cardiovascular dynamics unencumbered by central responses. PMID- 18407807 TI - Nanosecond electric pulse-induced calcium entry into chromaffin cells. AB - Electrically excitable bovine adrenal chromaffin cells were exposed to nanosecond duration electric pulses at field intensities ranging from 2 MV/m to 8 MV/m and intracellular calcium levels ([Ca(2+)](i)) monitored in real time by fluorescence imaging of cells loaded with Calcium Green. A single 4 ns, 8 MV/m pulse produced a rapid, short-lived increase in [Ca(2+)](i), with the magnitude of the calcium response depending on the intensity of the electric field. Multiple pulses failed to produce a greater calcium response than a single pulse, and a short refractory period was required between pulses before another maximal increase in [Ca(2+)](i) could be triggered. The pulse-induced rise in [Ca(2+)](i) was not affected by depleting intracellular calcium stores with caffeine or thapsigargin but was completely prevented by the presence of EGTA, Co(2+), or the L-type calcium channel blocker nitrendipine in the extracellular medium. Thus, a single nanosecond pulse is sufficient to elicit a rise in [Ca(2+)](i) that involves entry of calcium via L-type calcium channels. PMID- 18407808 TI - A note from the President of the Mitochondrial Medicine Society. PMID- 18407809 TI - High-throughput determination of fudosteine in human plasma by liquid chromatography-tandem mass spectrometry, following protein precipitation in the 96-well plate format. AB - A 96-well protein precipitation, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and fully validated for the determination of fudosteine in human plasma. After protein precipitation of the plasma samples (50 microL) by the methanol (150 microL) containing the internal standard (IS), erdosteine, the 96-well plate was vortexed for 5 min and centrifuged for 15 min. The 100 microL supernatant and 100 microL mobile phase were added to another plate and mixed and then the mixture was directly injected into the LC-MS/MS system in the negative ionization mode. The separation was performed on a XB-CN column for 3.0 min per sample using an eluent of methanol-water (60:40, v/v) containing 0.005% formic acid. Multiple reaction monitoring (MRM) using the precursor-product ion transitions m/z 178-->91 and m/z 284-->91 was performed to quantify fudosteine and erdosteine, respectively. The method was sensitive with a lower limit of quantification (LLOQ) of 0.02 microg mL(-1), with good linearity (r>0.999) over the linear range of 0.02-10 microg mL(-1). The within- and between run precision was less than 5.5% and accuracy ranged from 94.2 to 106.7% for quality control (QC) samples at three concentrations of 0.05, 1 and 8 microg mL( 1). The method was employed in the clinical pharmacokinetic study of fudosteine formulation product after oral administration to healthy volunteers. PMID- 18407810 TI - HPLC preparation of the chiral forms of 6-methoxy-gossypol and 6,6'-dimethoxy gossypol. AB - A concentrated mixture of gossypol, 6-methoxy-gossypol, and 6,6'-dimethoxy gossypol was extracted from the root bark of St. Vincent Sea Island cotton with acetone. This extract was derivatized with R-(-)-2-amino-1-propanol to form diastereomeric gossypol Schiff's bases. Analytical-scale reverse-phase chromatography of these Schiff's bases produced six peaks, indicating separation of the enantiomeric forms of the three gossypol compounds. The elution order of the peaks was found to vary with the polarity of the mobile phase. The chromatography was scaled to a preparative level and was used to isolate each compound. After hydrolysis of the separated Schiff's bases, the original compounds were recovered by precipitation from solutions of diethyl ether, acetic acid, and water. Fifty injections yielded approximately 500 mg of each methoxy gossypol enantiomer and 300 mg of each dimethoxy-gossypol enantiomer. Each compound was characterized for carbon and hydrogen content, optical rotation, UV vis light absorption, and melting point. Standard curves were developed and were used to measure the concentration of each gossypol form in the root bark and dehulled seed of St. Vincent Sea Island cotton. In seed tissue, 48% of the gossypol compounds were methylated, and the (-)-optical form was found to be in a slight excess to the (+)-optical form (53-54%) for all three compounds. In root bark, 71% of the gossypol compounds were methylated, and the (+)-optical form was in excess to the (-)-optical form for all three compounds. However, in this tissue the extent of enantiomeric excess decreased with the degree of methylation, with 77% of the gossypol existing in the (+)-optical form and 59% of the 6,6'-dimethoxy-gossypol existing in the (+)-optical form. PMID- 18407811 TI - Economic transformation and biological welfare in colonial Burma: regional differentiation in the evolution of average height. AB - Did economic development result in an improvement in biological welfare in the tropics before the diffusion of modern public health techniques in the 1950s and 1960s? Between the mid-19th and early 20th century, Lower Burma experienced a rapid rise in population and became increasingly commercialized as a major rice exporter. Land reclamation on a massive scale in the Irrawaddy delta required an arduous process of jungle clearance, land drainage and preparation, and canal and bund construction, mostly in malarial swamps. Once paddy lands were created, rice was grown with rudimentary tools in malarial zones. By contrast, in most parts of Upper Burma the economy remained more subsistence-oriented, and less commercialized. In this paper, we investigate changes in physical stature by processing and analyzing data reported in two anthropometric surveys conducted in various regions of Upper and Lower Burma in 1904 and in 1938-1941. An inverted U curve is observed in the evolution of average height in Lower Burma, while stature remained fairly stable in Upper Burma until the 1930s. PMID- 18407812 TI - Time, change and peptic ulcer disease in Rotherham, UK. AB - BACKGROUND AND AIM: We have documented the changing pattern of peptic ulcer disease in our centre in the last quarter of the 20th century and speculate on the reasons thereof. PATIENTS AND METHODS: The profile of peptic ulcer disease patients presenting newly to our centre (population 250,000) from 1977 to 2001 was examined. All patients were prospectively followed and detailed records kept. Results are presented in 5-year periods. RESULTS: Seven thousand five hundred and ninety new peptic ulcer disease patients (5564 duodenal ulcer+2026 gastric ulcer) were seen, peaking in 1982-1986 but declining thereafter, and with a falling male preponderance. Patients with gastric ulcer were older than those with duodenal ulcer; were older than duodenal ulcer, the mean age of both increased over time and the age gap from the general population widened. The numbers presenting with perforation changed little but haemorrhage increased, particularly amongst the elderly. Ulcers refractory to H2 receptor antagonists declined even before proton pump inhibitors were introduced. Elective surgery, already declining before H2 receptor antagonists, had virtually disappeared by 1992-1996. DISCUSSION AND CONCLUSION: Peptic ulcer disease affects an older population, an increasing proportion of whom present with haemorrhage. Refractoriness to H2 receptor antagonists and the need for elective operation was declining even before the emergence of modern treatment. We suggest the changes observed result not only from modern therapy but also substantially from a changing natural history. PMID- 18407813 TI - The inferior labial artery island flap. AB - The Abbe flap procedure has typically been indicated in cases of tissue defects of the upper lip after injury or tumour excision. However, this method requires two-stage reconstruction. In this report, we describe for the first time a novel one-stage reconstruction method using the inferior labial artery island flap. A 54-year-old man presented with a left upper lip defect and a scar contracture between the upper lip and the left cheek. We planned to reconstruct the lip defect using the inferior labial artery island flap. The inferior labial artery island flap was harvested with a vascular pedicle, and the vascular pedicle was returned through the inside of the flap. The flap survived completely, and liquid leakage from the lip and the appearance of the injured area were clearly improved. For this new technique, we converted the inferior labial flap to a vascular pedicled island flap, which increased its flexibility. This long vascular pedicle could be returned through the inside of the flap. Thus, this flap appears to be ideal for one-stage reconstructions of full-thickness upper lip defects. PMID- 18407814 TI - Modified method of shaving an avulsed scalp before replantation. PMID- 18407815 TI - Repair of recurrent midline abdominal wall dehiscence using the components separation technique. PMID- 18407816 TI - Response to: 'Malignant melanoma re-excision specimens: is there a need for histopathological analysis?' McGoldrick et al., J PRAS 2008; 61:983-984. PMID- 18407817 TI - Lower limb revascularisation preceding surgical wound coverage - an interdisciplinary algorithm for chronic wound closure. AB - BACKGROUND: Chronic wounds may occur or persist due to arterial insufficiency. Despite the high prevalence of arterial occlusive disease, a search of the literature has yielded a paucity of data on the benefit of interventions to recanalise lower extremity arteries prior to surgical closure of chronic wounds. OBJECTIVE: To investigate the correlation of simple clinical examinations and apparative diagnostics for the detection of arterial occlusive disease of the lower extremity in patients with chronic wounds, and to evaluate the benefit of vascular procedures to optimise wound perfusion before surgical closure. PATIENTS AND METHODS: During a 6-year period, 150 patients with chronic lower extremity wounds (no healing for more than 30 days) were included into this prospective study. All patients underwent palpation of foot pulses, Doppler sonography and measurement of occlusive pressures. Positive clinical findings were re-evaluated by angiography. All patients with peripheral extremity vessel occlusions underwent vascular interventions (percutaneous transluminal angioplasty with stenting, open thrombectomy or vascular bypass surgery) prior to surgical wound closure. RESULTS: In all 34 patients (21%) with missing foot pulses, suspicious Doppler signals or pathological occlusive pressure measurements, the clinical diagnosis of arterial occlusion was confirmed by angiography. An arterial pathology had previously been diagnosed in merely two of those patients. Nineteen patients underwent percutaneous transluminal angioplasty and 21 stents were implanted; in 10 cases, open thrombectomy or vascular bypasses were performed. In all 34 patients, sufficient peripheral recanalisation and improved wound perfusion were successfully achieved. For definitive wound closure, microsurgical tissue transplantation was performed in 15 patients. Angiography was performed prior to surgery. In 11 patients, regional or local flaps were used. Six patients received split skin grafting only; two wounds healed conservatively following vascular intervention. Vacuum-assisted closure (VAC) therapy was applied in 15 cases. Postoperatively, three cases of impaired wound healing and one infection occurred. CONCLUSIONS: Arterial insufficiency can be diagnosed safely by simple clinical examination. All clinically pathological results were successfully confirmed by angiography, allowing for a targeted peripheral vessel reopening to improve wound perfusion before surgery. This straightforward algorithm helped to improve the success of surgical therapy of chronic lower extremity wounds. PMID- 18407818 TI - A practical dressing to the umbilical stalk. PMID- 18407819 TI - Successful composite graft for fingertip amputations using ice-cooling and lipo prostaglandin E1. AB - In distal fingertip amputations where microanastomosis may not be possible, composite grafting might offer the possibility of maintaining digital length and function using the patient's own tissue. Many trials such as ice-cooling, pharmacologic enhancement and hyperbaric oxygenation have been reported to improve the survival rate of the composite graft. Twenty-four cases of unreplantable fingertip amputation were classified as types I to III according to the level of injury. We performed the composite grafting followed by immediate ice-cooling for 2 weeks and intravenous lipo-prostaglandin E(1) (lipo-PGE(1)) injection for 8.8 days each on average. Twenty-two fingertips in 24 patients survived completely with acceptable appearance and sensibility over the 8 month follow-up period. Confirming that therapeutic angiogenesis using ice-cooling and lipo-PGE(1) can increase the survival rate of the composite graft in unreplantable fingertip amputation, we describe the procedures and postoperative care in detail. PMID- 18407820 TI - Estrogen deficiency increases osteoclastogenesis up-regulating T cells activity: a key mechanism in osteoporosis. AB - Compelling evidences suggest that increased production of osteoclastogenic cytokines by activated T cells plays a relevant role in the bone loss induced by estrogen deficiency in the mouse. However, little information is available on the role of T cells in post-menopausal bone loss in humans. To investigate this issue we have assessed the production of cytokines involved in osteoclastogenesis (RANKL, TNFalpha and OPG), in vitro osteoclast (OC) formation in pre and post menopausal women, the latter with or without osteoporosis. We evaluated also OC precursors in peripheral blood and the ability of peripheral blood mononuclear cells to produce TNFalpha in both basal and stimulated condition by flow cytometry in these subjects. Our data demonstrate that estrogen deficiency enhances the production of the pro-osteoclastogenetic cytokines TNFalpha and RANKL and increases the number of circulating OC precursors. Furthermore, we show that T cells and monocytes from women with osteoporosis exhibit a higher production of TNFalpha than those from the other two groups. Our findings suggest that estrogen deficiency stimulates OC formation both by increasing the production of TNFalpha and RANKL and increasing the number of OC precursors. Women with post-menopausal osteoporosis have a higher T cell activity than healthy post-menopausal subjects; T cells thus contribute to the bone loss induced by estrogen deficiency in humans as they do in the mouse. PMID- 18407821 TI - The study of abnormal bone development in the Apert syndrome Fgfr2+/S252W mouse using a 3D hydrogel culture model. AB - Apert syndrome is caused by mutations in fibroblast growth factor receptor 2 (Fgfr2) and is characterized by craniosynostosis and other skeletal abnormalities. The Apert syndrome Fgfr2+/S252W mouse model exhibits perinatal lethality. A 3D hydrogel culture model, derived from tissue engineering strategies, was used to extend the study of the effect of the Fgfr2+/S252W mutation in differentiating osteoblasts postnatally. We isolated cells from the long bones of Apert Fgfr2+/S252W mice (n=6) and cells from the wild-type sibling mice (n=6) to be used as controls. During monolayer expansion, Fgfr2+/S252W cells demonstrated increased proliferation and ALP activity, as well as altered responses of these cellular functions in the presence of FGF ligands with different binding specificity (FGF2 or FGF10). To better mimic the in vivo disease development scenario, cells were also encapsulated in 3D hydrogels and their phenotype in 3D in vitro culture was compared to that of in vivo tissue specimens. After 4 weeks in 3D culture in osteogenic medium, Fgfr2+/S252W cells expressed 2.8-fold more collagen type I and 3.3-fold more osteocalcin than did wild-type controls (p<0.01). Meanwhile, Fgfr2+/S252W cells showed decreased bone matrix remodeling and expressed 87% less Metalloprotease-13 and 71% less Noggin (p<0.01). The S252W mutation also led to significantly higher production of collagen type I and II in 3D as shown by immunofluorescence staining. In situ hybridization and alizarin red S staining of postnatal day 0 (P0) mouse limb sections demonstrated significantly higher levels of osteopontin expression and mineralization in Fgfr2+/S252W mice. Complementary to in vivo findings, this 3D hydrogel culture system provides an effective in vitro venue to study the pathogenesis of Apert syndrome caused by the analogous mutation in humans. PMID- 18407822 TI - Spatial distribution of anisotropic acoustic impedance assessed by time-resolved 50-MHz scanning acoustic microscopy and its relation to porosity in human cortical bone. AB - We used quantitative scanning acoustic microscopy (SAM) to assess tissue acoustic impedance and microstructure of cortical bone of human radii with the aim to provide data on regional distribution of acoustic impedance along the circumferential and across the radial directions in the entire cross-section of the radius diaphysis as well as to determine the range of impedance values in transverse (perpendicular to bone axis) and longitudinal (parallel to bone axis) cross-sections. Several microstructural features related to cortical porosity were analyzed in order to determine whether these features differ in different parts of the cortex and to assess the relationship between the microstructure and tissue acoustic impedance. Fifteen fresh bone specimens (human radius) were investigated using a SAM (center frequency of 50 MHz and -6 dB lateral resolution of approximately 23 microm). The sample acoustic impedance was obtained by means of a calibration curve correlating the reflected signal amplitude of reference materials with their corresponding well-known acoustic impedance. Tissue acoustic impedance and microstructural features were derived from the morphometric analysis of the segmented impedance images. A higher porosity was found in the inner cortical layer (mean+/-SD=8.9+/-2.3%) compared to the peripheral layer (2.7+/-1.5%) (paired t-test, p<10(-5)). ANOVA showed that most of the variance can be explained by the regional effect across the radial direction with a minor contribution due to between-sample variability. Similar to porosity, the number and diameter of pores were greater in the inner layer. In contrast to porosity, ANOVA showed that impedance variability can mostly be explained by between specimen variability. Two-way ANOVA revealed that after compensation for the between-sample variability the variation in acoustic impedance across the radial direction was much larger than that along the circumferential direction. In addition to the significant difference between the inner cortical layer (8.25+/ 0.4 Mrayl) and peripheral layer (8.0+/-0.5 Mrayl) (unilateral paired t-test, p<10(-4)), the values in the anterior region (8.2+/-0.5 Mrayl) were found to be significantly higher than those of the posterior region (7.9+/-0.6 Mrayl). Impedance mean value of longitudinal sections was lower than mean value measured in transverse cross-sections, resulting in an impedance acoustic anisotropy ratio of 1.17+/-0.03 in the inner cortical layer and 1.19+/-0.02 in the peripheral layer. SAM is a valuable tool to provide data on the spatial distribution of microstructural and microelastic bone properties that is useful to improve our understanding of the impact of bone microstructure on tissue material properties. PMID- 18407824 TI - A pyrroloquinoline quinine-dependent membrane-bound d-sorbitol dehydrogenase from Gluconobacter oxydans exhibits an ordered Bi Bi reaction mechanism. AB - A membrane-bound pyrroloquinoline quinine (PQQ)-dependent D-sorbitol dehydrogenase (mSLDH) in Gluconobacter oxydans participates in the oxidation of D sorbitol to L-sorbose by transferring electrons to ubiquinone which links to the respiratory chain. To elucidate the kinetic mechanism, the enzyme purified was subjected to two-substrate steady-state kinetic analysis, product and substrate inhibition studies. These kinetic data indicate that the catalytic reaction follows an ordered Bi Bi mechanism, where the substrates bind to the enzyme in a defined order (first ubiquinone followed by D-sorbitol), while products are released in sequence (first L-sorbose followed by ubiquinol). From these findings, we proposed that the native mSLDH bears two different substrate-binding sites, one for ubiquinone and the other for D-sorbitol, in addition to PQQ binding and Mg(2+)-binding sites in the catalytic center. PMID- 18407825 TI - Recent progress in elucidating the molecular mechanism of the mitochondrial permeability transition pore. AB - The mitochondrial permeability transition pore (MPTP) plays a key role in cell death, especially necrosis, and mediates the injury tissues such as the heart and brain experience following ischaemia and reperfusion. However, the molecular identity of the MPTP remains uncertain. Knockout studies have confirmed a role for cyclophilin-D (CyP-D) in pore opening, probably mediated by its peptidyl prolyl cis-trans isomerase activity that facilitates a conformational change in an inner membrane protein. However, similar knockout studies have cast doubt on the central role of the adenine nucleotide translocase (ANT), previously regarded as a leading contender for the membrane component that forms the transmembrane channel of the MPTP. Here we review the evidence for and against a role for the ANT in MPTP opening and conclude that it usually plays a regulatory role rather than provide the transmembrane pore component. We suggest that the protein fulfilling the latter role is the mitochondrial phosphate carrier (PiC) and summarise recent evidence in support of this proposal. Our data are consistent with a model for the MPTP in which a calcium-triggered conformational change of the PiC, facilitated by CyP-D, induces pore opening. We propose that this is enhanced by an association of the PiC with the "c" conformation of the ANT. Agents that modulate pore opening may act on either or both the PiC and the ANT. PMID- 18407826 TI - Binding and interactions of L-BABP to lipid membranes studied by molecular dynamic simulations. AB - Chicken liver bile acid-binding protein (L-BABP) is a member of the fatty acid binding proteins super family. The common fold is a beta-barrel of ten strands capped with a short helix-loop-helix motif called portal region, which is involved in the uptake and release of non-polar ligands. Using multiple-run molecular dynamics simulations we studied the interactions of L-BABP with lipid membranes of anionic and zwitterionic phospholipids. The simulations were in agreement with our experimental observations regarding the electrostatic nature of the binding and the conformational changes of the protein in the membrane. We observed that L-BABP migrated from the initial position in the aqueous bulk phase to the interface of anionic lipid membranes and established contacts with the head groups of phospholipids through the side of the barrel that is opposite to the portal region. The conformational changes in the protein occurred simultaneously with the binding to the membrane. Remarkably, these conformational changes were observed in the portal region which is opposite to the zone where the protein binds directly to the lipids. The protein was oriented with its macrodipole aligned in the configuration of lowest energy within the electric field of the anionic membrane, which indicates the importance of the electrostatic interactions to determine the preferred orientation of the protein. We also identified this electric field as the driving force for the conformational change. For all the members of the fatty acid-binding protein family, the interactions with lipid membranes is a relevant process closely related to the uptake, release and transfer of the ligand. The observations presented here suggest that the ligand transfer might not necessarily occur through the domain that directly interacts with the lipid membrane. The interactions with the membrane electric field that determine orientation and conformational changes described here can also be relevant for other peripheral proteins. PMID- 18407827 TI - Hard tissue regeneration capacity of apical pulp derived cells (APDCs) from human tooth with immature apex. AB - Recent studies indicate that dental pulp is a new source of adult stem cells. The human tooth with an immature apex is a developing organ, and the apical pulp of this tooth may contain a variety of progenitor/stem cells, which participate in root formation. We investigated the hard tissue regeneration potential of apical pulp derived cells (APDCs) from human tooth with an immature apex. APDCs cultured with a mineralization-promoting medium showed alkaline phosphatase activity in porous hydroxyapatite (HA) scaffolds. The composites of APDCs and HA were implanted subcutaneously in immunocompromised rats and harvested at 12 weeks after implantation. In histological analysis, the APDCs/HA composites exhibited bone- and dentine-like mineralized tissues in the pore areas of HA. This study suggests that the human tooth with an immature apex is an effective source of cells for hard tissue regeneration. PMID- 18407828 TI - TRAF6-mediated regulation of the PI3 kinase (PI3K)-Akt-GSK3beta cascade is required for TNF-induced cell survival. AB - We recently demonstrated that the tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) helps maintenance of cell survival by regulating glycogen synthase kinase 3beta (GSK3beta) activity during TNF signaling. However, the molecular linkage between TRAF6 and GSK3beta signaling is unknown. Herein, we showed that TRAF6 positively regulated cell survival by modulating PI3K-Akt GSK3beta cascades. In 3T3 cells lacking TRAF6, but not those lacking TRAF2, TNF stimulation led to prolonged hyperphosphorylation of Akt, which coincided with the activation of upstream PI3K. Pharmacologically blocking PI3K significantly inhibited Akt and GSK3beta phosphorylation. Importantly, PI3K inhibition rescued cell death in TRAF6-null 3T3 cells. These data suggested TRAF6 regulates TNF mediated cell survival through PI3K-Akt-GSK3beta cascades. PMID- 18407829 TI - Role of the hinge region of glucocorticoid receptor for HEXIM1-mediated transcriptional repression. AB - We previously reported that HEXIM1 (hexamethylene bisacetamide-inducible protein 1), which suppresses transcription elongation via sequestration of positive transcription elongation factor b (P-TEFb) using 7SK RNA as a scaffold, directly associates with glucocorticoid receptor (GR) to suppress glucocorticoid-inducible gene activation. Here, we revealed that the hinge region of GR is essential for its interaction with HEXIM1, and that oxosteroid receptors including GR show sequence homology in their hinge region and interact with HEXIM1, whereas the other members of nuclear receptors do not. We also showed that HEXIM1 suppresses GR-mediated transcription in two ways: sequestration of P-TEFb by HEXIM1 and direct interaction between GR and HEXIM1. In contrast, peroxisome proliferator activated receptor gamma-dependent gene expression is negatively modulated by HEXIM1 solely via sequestration of P-TEFb. We, therefore, conclude that HEXIM1 may act as a gene-selective transcriptional regulator via direct interaction with certain transcriptional regulators including GR and contribute to fine-tuning of, for example, glucocorticoid-mediated biological responses. PMID- 18407830 TI - PLAP-1/asporin inhibits activation of BMP receptor via its leucine-rich repeat motif. AB - We previously identified the novel gene, periodontal ligament-associated protein 1 (PLAP-1)/asporin and reported that PLAP-1/asporin inhibited bone morphogenetic protein-2 (BMP-2)-induced cytodifferentiation of periodontal ligament (PDL) cells probably by direct interaction with BMP-2. Here, we elucidated the detailed regulatory mechanism of this protein on BMP-2-induced cytodifferentiation of PDL cells. Recombinant PLAP-1/asporin inhibited BMP-2-induced cytodifferentiation of PDL cells and competitively prevented BMP-2 from binding to the BMP receptor-IB (BMPR-IB), resulting in inhibition of BMP-dependent activation of Smad proteins. The induction of mutation to the leucine-rich repeat (LRR) motif, especially LRR5, within PLAP-1/asporin rescued the inhibitory effect of PLAP-1/asporin on BMP-2. By contrast, a 26-amino acid peptide in the PLAP-1/asporin LRR5 sequence inhibited BMP-2 activity. Our findings indicate that PLAP-1/asporin inhibits BMP 2-induced differentiation of PDL cells resulting from inactivation of the BMP-2 signaling pathway and that LRR, especially LRR5 of PLAP-1/asporin, plays an important role in the PLAP-1/asporin-BMP-2 interaction. PMID- 18407831 TI - Nano titanium dioxide photocatalytic protein tyrosine nitration: a potential hazard of TiO2 on skin. AB - Protein tyrosine nitration is a prevalent post-translational modification which occurs as a result of oxidative and nitrative stress, it may be directly involved in the onset and/or progression of diseases. Considering the existence of nano titanium dioxide (TiO(2)) in environment and sunscreen products along with the high content of nitrite in sweat, the UV-exposed skin may be a significant target for the photosensitized damage. In this paper, tyrosine nitration of bovine serum albumin (BSA) was initiated in the UV-irradiated reaction mixture containing 0.2 3.0mg/ml of three commercially nano TiO(2) products and 0.25-1.0mM NO2-. It was found that anatase TiO(2) and Degussa P25 TiO(2) showed prominent photocatalytic activity on promoting the formation of protein tyrosine nitration, and the optimum condition for the reaction was around physiological pH. Meanwhile, the photocatalytic effect of rutile on protein tyrosine nitration was subtle. The potential physiological significance of nano TiO(2)-photocatalytic protein nitration was also demonstrated in mouse skin homogenate. Although the relationship between photocatalytic protein tyrosine nitration and chronic cutaneous diseases needs further study, the toxicity of nano TiO(2) to the skin disease should be paid more attention in the production and utilization process. PMID- 18407832 TI - Delay expression of limonoid UDP-glucosyltransferase makes delayed bitterness in citrus. AB - Genes encoding limonoid UDP-glucosyltransferase from albedo of six Citrus species with different levels of delayed bitterness are isolated and cloned in vector pTZ57R/T. Our results indicate that gene sequence of sweet lime (with intense juice delayed bitterness) have complete identity with Satsuma mandarin (without distinctive juice delayed bitterness). Also gene sequence of Marsh seedless grapefruit, local orange and Thompson navel orange (with mild juice delayed bitterness) have very similarity with Satsuma mandarin. On the other hand, this gene started to express 60, 120, and 210 days after full blooming in albedo of Satsuma mandarin, sweet oranges and sour orange, and both grapefruit and sweet lime, respectively. Expression pattern of limonoid glucosyltransferase gene in leaves was quite different with albedo. Thus, we supposed the delayed bitterness in this species was related to delay in expression of limonoid glucosyltransferase gene in albedo and lower limonoid glucoside accumulation in fruits. PMID- 18407833 TI - Reactive oxygen species generated by thiol-modifying phenylarsine oxide stimulate the expression of protein L-isoaspartyl methyltransferase. AB - Expression of the repair enzyme protein L-isoaspartyl methyltransferase (PIMT) has been reported to play important roles in brain. However, little is known about the regulation of PIMT expression following protein damage by oxidation in brain. Phenylarsine oxide (PAO) is an arsenical compound that alters proteins by forming disulfide bond with vicinal cysteinyl residues. Here we report that PIMT was rapidly up-regulated by PAO in U-87 human astroglioma cells. We also confirmed that PIMT up-regulation by PAO was mediated by the reaction with vicinal cysteines. Furthermore, we showed that PIMT induction by PAO was dependent on formation of reactive oxygen species (ROS). Crucially, both ROS formation and PIMT induction by PAO were inhibited by antioxidant N-acetyl-L cysteine and NADPH oxidase inhibitor diphenyleneiodonium chloride. Importantly, down-regulation of PIMT by siRNA strikingly enhanced PAO-induced ROS. Together, these results highlight that PIMT expression is regulated by ROS and could primarily act as an antioxidant enzyme. PMID- 18407834 TI - Impact of free hydroxylated and methyl-branched fatty acids on the organization of lipid membranes. AB - Differential scanning calorimetry (DSC) has been applied to study the effect of free hydroxylated and methyl-branched fatty acids on the physico-chemical properties of lipid membranes. First, the impact of free hydroxy fatty acids (HFAs) on dimyristoylphosphatidylcholine (DMPC) model membranes was monitored only as a function of chain length and position of the attached hydroxyl group. Second, racemic vs. enantiopure anteiso fatty acids (AFAs) and HFAs were investigated to address the question of which role does a fatty acid's chirality play on its membrane pertubing effect. The DSC thermograms revealed that the main gel to liquid-crystalline phase transition of the DMPC bilayers which results in a disordering effect of the lipid hydrocarbon chains was affected in different ways depending on the nature of the incorporated fatty acid. Long-chain 2- and 3 HFAs stabilized the gel phase by reducing the phase transition temperature (T(m)), whereas short-chain HFAs and long-chain HFAs with the hydroxy group remote from the head group stabilized the more disordered liquid-crystalline state. Additionally, we observed that enantiopure (S)-14-methylhexadecanoic acid ((S)-a17:0) and (R)-2-hydroxy octadecanoic acid and the corresponding racemates had contrary effects upon incorporation into DMPC bilayers. In both cases, the pure enantiomers alleviated the liquid-crystalline state of the biological model membrane. PMID- 18407835 TI - Phase behaviour of binary mixtures involving tristearin, stearyl stearate and stearic acid: thermodynamic study and BAM observation at the air-water interface and AFM analysis of LB films. AB - The binary mixtures involving tristearin (TS), stearyl stearate (SS) and stearic acid (SA) were studied by surface pressure-area (pi-A) measurements and by Brewster angle microscopy (BAM), at the air-water interface, and the Langmuir Blodgett (LB) monolayers, transferred onto mica substrates, were analysed by AFM. The thermodynamic analysis indicated miscibility in the whole composition range for the system SA/TS, and partial miscibility for systems SA/SS and TS/SS. This behaviour was further confirmed by BAM observation and AFM analysis of LB films. The AFM imaging of collapsed monolayers revealed domains with a multilayered structure varying with system and composition. The layers thickness determined by cross section analysis are consistent with estimated molecular lengths and conformations proposed for the molecules, assuming nearly perpendicular or tilted orientations of the hydrocarbon chains to the interface. PMID- 18407836 TI - The effect of propoxycaine.HCl on the physical properties of neuronal membranes. AB - Fluorescent probe techniques were used to evaluate the effect of propoxycaine.HCl on the physical properties (transbilayer asymmetric lateral and rotational mobilities, annular lipid fluidity and protein distribution) of synaptosomal plasma membrane vesicles (SPMVs) isolated from bovine cerebral cortex. An experimental procedure was used based on selective quenching of both 1,3-di(1 pyrenyl)propane (Py-3-Py) and 1,6-diphenyl-1,3,5-hexatriene (DPH) by trinitrophenyl groups, and radiationless energy transfer (RET) from the tryptophans of membrane proteins to Py-3-Py. Propoxycaine.HCl increased the bulk lateral and rotational mobilities, and annular lipid fluidity in SPMVs lipid bilayers, and had a greater fluidizing effect on the inner monolayer than that of the outer monolayer. The magnitude of increasing effect on annular lipid fluidity in SPMVs lipid bilayer induced by propoxycaine.HCl was significantly far greater than magnitude of increasing effect of the drug on the lateral and rotational mobilities of SPMVs lipid bilayer. It also caused membrane proteins to cluster. These effects of propoxycaine.HCl on neuronal membranes may be responsible for some, though not all, of the local anesthetic actions of propoxycaine.HCl. PMID- 18407837 TI - Studies of biochemistry and clinical biochemistry. Studies at sample medical schools in 13 EU countries regarding biochemistry and clinical biochemistry teaching. AB - The study summarizes the results obtained during personal visits to 53 medical schools in the 13 original EU countries during 2004--2006. Data from the Czech Republic is shown for comparison. The possibilities of acquiring information from the websites of the medical schools in the local language and English are assessed. The admission process to medical schools and the organization of studies of medicine, dentistry, and non-medical healthcare fields are briefly characterized. Significant attention is paid to the forms of education in biochemistry and clinical (bio)chemistry in the medical study field. The position of these subjects in the studies of dentistry and non-medical healthcare fields is also noted. In addition, the course of subject exams is described. The methods of funding and postgraduate studies at the medical schools are also briefly addressed. PMID- 18407838 TI - The new Italian course of post-graduate education in laboratory medicine. AB - Clinical biochemistry has undergone a dramatic transformation in the last decade, with the introduction of new technologies, instrumentation and methodologies. The educational program of the postgraduate training should allow the students to achieve knowledge and skills to work and supervise a laboratory department in a hospital or health service institution. Here we describe the proposed revision of the postgraduate training in the Italian system, focusing on educational goals as well as mandatory professional activities. PMID- 18407839 TI - Evaluation of the Bayer microalbumin/creatinine urinalysis dipstick. AB - BACKGROUND: The objective of this study was to evaluate the utility of the Bayer microalbumin/creatinine urine reagent strip compared to established laboratory methods. METHODS: Random urine specimens from low and high risk pregnancy clinics as well as a random cohort from the community were analyzed for microalbumin and creatinine using both the reagent strip and the Roche Integra 700, according to manufacturers' specifications. Sensitivity and specificity were then calculated. RESULTS: For the pregnant cohorts the sensitivities ranged from 19% to 59%, and the range of specificities was 45.4% to 84.2%. Using the microalbumin/creatinine data from the community, the sensitivity and specificity of the strip were 52.4% and 97.6%, respectively. CONCLUSION: The poor sensitivity of the microalbumin/creatinine urine reagent strip to detect significant microalbuminuria will likely limit its usefulness as a screening tool. PMID- 18407840 TI - Education and training of medical technologists in Italy. AB - The role and figure of biomedical laboratory technologists have undergone important changes over the past decades. The increasingly complex functions and responsibility of biomedical laboratory technologists both require an updated education and training process. Here, we describe the current situation in Italy regarding the education of biomedical laboratory technologists and the results of a survey conducted in 28 universities. A significant variability between the different universities was observed regarding both the teaching subjects and the distribution of credits among the different courses. The results of this survey highlight that a greater uniformity in the distribution of credits and a better balance among teaching sectors might be advisable. PMID- 18407841 TI - The ER-associated degradation component Der1p and its homolog Dfm1p are contained in complexes with distinct cofactors of the ATPase Cdc48p. AB - Misfolded proteins in the endoplasmic reticulum (ER) are often degraded in the cytosol by a process called ER-associated protein degradation (ERAD). During ERAD in S. cerevisiae, the ATPase Cdc48p associates with Der1p, a putative component of a retro-translocation channel. Cdc48p also binds a homolog of Der1p, Dfm1p, that has no known function in ERAD. Here, we show that Der1p and Dfm1p are contained in distinct complexes. While the complexes share several ERAD components, only the Dfm1p complex contains the Cdc48p cofactors Ubx1p and Ubx7p, while the Der1p complex is enriched in Ufd1p. These data suggest distinct functions for the Der1p and Dfm1p complexes. PMID- 18407844 TI - Dynamic in-plane resonant characteristics of piezoceramic and piezolaminated composite plates. AB - Piezolaminated composite plates have received considerable attention in various industrial applications due to their intelligent characteristics. In this investigation, two experimental measurement techniques are used to determine the in-plane resonant vibration of angle-ply laminated composites embedded with a piezoceramic layer (piezolaminated plates) for different stacking angles. The first method is a full-field optical technique, which is called the AF-ESPI (amplitude-fluctuation electronic speckle pattern interferometry). This is the major experimental method. The AF-ESPI method is used to determine the in-plane resonant frequency and corresponding mode shape of a single-layer piezoceramic plate and piezolaminated plates with five different stacking angles. The second experimental technique, the impedance analyzer, is employed to determine the in plane resonant frequency. Finally, numerical computations based on the finite element analysis are presented for comparison of the two experimental results. Excellent agreement between the experimentally measured data and the numerically calculated results are found for in-plane resonant frequencies and mode shapes. This study indicates that the dynamic characteristics of inplane resonant vibrations for piezolaminated plates with different stacking angles are quite different. PMID- 18407845 TI - Pyroelectric energy conversion: optimization principles. AB - In the framework of microgenerators, we present in this paper the key points for energy harvesting from temperature using ferroelectric materials. Thermoelectric devices profit from temperature spatial gradients, whereas ferroelectric materials require temporal fluctuation of temperature, thus leading to different applications targets. Ferroelectric materials may harvest perfectly the available thermal energy whatever the materials properties (limited by Carnot conversion efficiency) whereas thermoelectric material's efficiency is limited by materials properties (ZT figure of merit). However, it is shown that the necessary electric fields for Carnot cycles are far beyond the breakdown limit of bulk ferroelectric materials. Thin films may be an excellent solution for rising up to ultra-high electric fields and outstanding efficiency. Different thermodynamic cycles are presented in the paper: principles, advantages, and drawbacks. Using the Carnot cycle, the harvested energy would be independent of materials properties. However, using more realistic cycles, the energy conversion effectiveness remains dependent on the materials properties as discussed in the paper. A particular coupling factor is defined to quantify and check the effectiveness of pyroelectric energy harvesting. It is defined similarly to an electromechanical coupling factor as k2=p2theta0/(epsilontheta33cE), where p, theta0, epsilontheta33, cE are pyroelectric coefficient, maximum working temperature, dielectric permittivity, and specific heat, respectively. The importance of the electrothermal coupling factor is shown and discussed as an energy harvesting figure of merit. It gives the effectiveness of all techniques of energy harvesting (except the Carnot cycle). It is finally shown that we could reach very high efficiency using 1110.75Pb(Mg1/3Nb2/3)-0.25PbTiO3 single crystals and synchronized switch harvesting on inductor (almost 50% of Carnot efficiency). Finally, practical implementation key points of pyroelectric energy harvesting are presented showing that the different thermodynamic cycles are feasible and potentially effective, even compared to thermoelectric devices. PMID- 18407842 TI - Biological roles of lysophospholipid receptors revealed by genetic null mice: an update. AB - Two lysophospholipids (LPs), lysophosphatidic acid (LPA) and sphingosine 1 phosphate (S1P), are known to affect various cellular events. Their actions are mediated by binding to at least ten bona fide high-affinity G protein-coupled receptors referred to as LPA1-5 and S1P1-5. These LPs are expressed throughout the body and are involved in a range of biological activities including normal development, as well as functioning in most organ systems. A growing number of biological functions have been uncovered in vivo using single- or multiple-null mice for each LP receptor. This review will focus on findings from in vivo as well as in vitro studies using genetic null mice for the LP receptors, LPA1,2,3 and S1P1,2,3,5, and for the LP producing enzymes, autotaxin and sphingosine kinase 1/2. PMID- 18407846 TI - Anomalous elastic properties of RF-sputtered amorphous TeO2+x thin film for temperature-stable SAW device applications. AB - The anomalous elastic properties of TeO2+x thin films deposited by rf diode sputtering on substrates at room temperature have been studied. The deposited films are amorphous, and IR spectroscopy reveals the formation of Te-O bond. X ray photoelectron spectroscopy confirms the variation in the stoichiometry of TeO2+x film from x=0 to 1 with an increase in the oxygen percentage in processing gas composition. The elastic parameters of the films in comparison to the reported values for TeO2+x single crystal are found to be low. However, the temperature coefficients of elastic parameters of all deposited films exhibit anomalous behavior showing positive values for TC(C11) in the range (32.0 to 600.0)x10(-4) degrees C(-1) and TC(C44)=(35.0 to 645.5)x10(-4) degrees C(-1) against the negative values TC(C11)=-2.7x10(-4) degrees C(-1) and TC(C44)= 0.73x10(-4) degrees C(-1) reported for TeO2+x single crystal. The variation in the elastic parameters and their temperature coefficients is correlated with the change in the three-dimensional network of Te-O bonding. The anomalous elastic properties of the TeO2+x films grown in 100% O2 are useful for potential application in the design of temperature stable surface acoustic wave devices. PMID- 18407843 TI - Interspecies regulation of microRNAs and their targets. AB - MicroRNAs (miRNAs) are 20-24 nucleotide RNA molecules that play essential roles in posttranscriptional regulation of target genes. In animals, miRNAs bind to target mRNA through imperfect complementary sequences that are usually located at the 3' untranslated regions (UTRs), leading to translational repression or transcript degradation. In plants, miRNAs predominately mediate degradation of target mRNAs via perfect or near-perfect complementary sequences. MicroRNA targets include a large number of transcription factors, suggesting a role of miRNAs in the control of regulatory networks and cellular growth and development. Many miRNAs and their targets are conserved among plants or animals, whereas some are specific to a few plant or animal lineages. Conserved miRNAs do not necessarily exhibit the same expression levels or patterns in different species or at different stages within a species. Therefore, sequence and expression divergence in miRNAs between species may affect miRNA accumulation and target regulation in interspecific hybrids and allopolyploids that contain two or more divergent genomes, leading to developmental changes and phenotypic variation in the new species. PMID- 18407847 TI - Single-ensemble-based eigen-processing methods for color flow imaging--Part I. The Hankel-SVD filter. AB - Because of their adaptability to the slow-time signal contents, eigen-based filters have shown potential in improving the flow detection performance of color flow images. This paper proposes a new eigen-based filter called the Hankel-SVD filter that is intended to process each slowtime ensemble individually. The new filter is derived using the notion of principal Hankel component analysis, and it achieves clutter suppression by retaining only the principal components whose order is greater than the clutter eigen-space dimension estimated from a frequency based analysis algorithm. To assess its efficacy, the Hankel-SVD filter was first applied to synthetic slow-time data (ensemble size: 10) simulated from two different sets of flow parameters that model: 1) arterial imaging (blood velocity: 0 to 38.5 cm/s, tissue motion: up to 2 mm/s, transmit frequency: 5 MHz, pulse repetition period: 0.4 ms) and 2) deep vessel imaging (blood velocity: 0 to 19.2 cm/s, tissue motion: up to 2 cm/s, transmit frequency: 2 MHz, pulse repetition period: 2.0 ms). In the simulation analysis, the post-filter clutter to- blood signal ratio (CBR) was computed as a function of blood velocity. Results show that for the same effective stopband size (50 Hz), the Hankel-SVD filter has a narrower transition region in the post-filter CBR curve than that of another type of adaptive filter called the clutter-downmixing filter. The practical efficacy of the proposed filter was tested by application to in vivo color flow data obtained from the human carotid arteries (transmit frequency: 4 MHz, pulse repetition period: 0.333 ms, ensemble size: 10). The resulting power images show that the Hankel-SVD filter can better distinguish between blood and moving-tissue regions (about 9 dB separation in power) than the clutter downmixing filter and a fixed-rank multi ensemble-based eigen-filter (which showed a 2 to 3 dB separation). PMID- 18407848 TI - Single-ensemble-based eigen-processing methods for color flow imaging--Part II. The matrix pencil estimator. AB - Parametric spectral estimators can potentially be used to obtain flow estimates directly from raw slow-time ensembles whose clutter has not been suppressed. We present a new eigen-based parametric flow estimation method called the matrix pencil, whose principles are based on a matrix form under the same name. The presented method models the slow-time signal as a sum of dominant complex sinusoids in the slow-time ensemble, and it computes the principal Doppler frequencies by using a generalized eigen-value problem-formulation and matrix rank reduction principles. Both fixed rank (rank-one, rank-two) and adaptive-rank matrix pencil flow estimators are proposed, and their potential applicability to color flow signal processing is discussed. For the adaptive-rank estimator, the nominal rank was defined as the minimum eigen-structure rank that yields principal frequency estimates with a spread greater than a prescribed bandwidth. In our initial performance evaluation, the fixed-rank matrix pencil estimators were applied to raw color flow data (transmit frequency: 5 MHz; pulse repetition period: 0.175 ms; ensemble size: 14) acquired from a steady flow phantom (70 cm/s at centerline) that was surrounded by rigid-tissue-mimicking material. These fixed-rank estimators produced velocity maps that are well correlated with the theoretical flow profile (correlation coefficient: 0.964 to 0.975). To facilitate further evaluation, the matrix pencil estimators were applied to synthetic slow time data (transmit frequency: 5 MHz; pulse repetition period: 1.0 ms; ensemble size: 10) modeling flow scenarios without and with tissue motion (up to 1 cm/s). The bias and root-mean-squared error of the estimators were computed as a function of blood-signal-to-noise ratio and blood velocity. The matrix pencil flow estimators showed that they are comparatively less biased than most of the existing frequency-based flow estimators like the lagone autocorrelator. PMID- 18407849 TI - The cross algorithm for phase-aberration correction in medical ultrasound images formed with two-dimensional arrays. AB - Common-midpoint signals in the near-field signal-redundancy (NFSR) algorithm for one-dimensional arrays are acquired using three consecutive transducer elements. An all-row-plus-two-column algorithm has been proposed to implement the one dimensional NFSR algorithm on two dimensional arrays. The disadvantage of this method is that its ambiguity profile is not linear and a timeconsuming iterative method has to be used to linearize the ambiguity profile. An all-row-plus-two column-and-a-diagonal algorithm has also been proposed. Its ambiguity profile is linear, but it is very sensitive to noise and cannot be used. In this paper, a novel cross algorithm is proposed to implement the NFSR algorithm on two dimensional arrays. In this algorithm, common-midpoint signals are acquired using four adjacent transducer elements, which is not available in one-dimensional arrays. Its advantage includes a linear ambiguity profile and a higher measurement signal-to-noise ratio. The performance of the cross algorithm is evaluated theoretically. The region of redundancy is analyzed. The procedure for deriving the phaseaberration profile from peak positions of cross-correlation functions between common-midpoint signals is discussed. This algorithm is tested with a simulated data set acquired with a two-dimensional array, and the result shows that the cross algorithm performs better than the all-row plus-twocolumn NFSR algorithm. PMID- 18407850 TI - The acoustic lens design and in vivo use of a multifunctional catheter combining intracardiac ultrasound imaging and electrophysiology sensing. AB - A multifunctional 9F intracardiac imaging and electrophysiology mapping catheter was developed and tested to help guide diagnostic and therapeutic intracardiac electrophysiology (EP) procedures. The catheter tip includes a 7.25-MHz, 64 element, side-looking phased array for high resolution sector scanning. Multiple electrophysiology mapping sensors were mounted as ring electrodes near the array for electrocardiographic synchronization of ultrasound images. The catheter array elevation beam performance in particular was investigated. An acoustic lens for the distal tip array designed with a round cross section can produce an acceptable elevation beam shape; however, the velocity of sound in the lens material should be approximately 155 m/s slower than in tissue for the best beam shape and wide bandwidth performance. To help establish the catheter's unique ability for integration with electrophysiology interventional procedures, it was used in vivo in a porcine animal model, and demonstrated both useful intracardiac echocardiographic visualization and simultaneous 3-D positional information using integrated electroanatomical mapping techniques. The catheter also performed well in high frame rate imaging, color flow imaging, and strain rate imaging of atrial and ventricular structures. PMID- 18407851 TI - Capon beamforming in medical ultrasound imaging with focused beams. AB - Medical ultrasound imaging is conventionally done by insonifying the imaged medium with focused beams. The backscattered echoes are beamformed using delay and-sum operations that cannot completely eliminate the contribution of signals backscattered by structures off the imaging beam to the beamsum. It leads to images with limited resolution and contrast. This paper presents an adaptation of the Capon beamformer algorithm to ultrasound medical imaging with focused beams. The strategy is to apply data-dependent weight functions to the imaging aperture. These weights act as lateral spatial filters that filter out off-axis signals. The weights are computed for each point in the imaged medium, from the statistical analysis of the signals backscattered by that point to the different elements of the imaging probe when insonifying it with different focused beams. Phantom and in vivo images are presented to illustrate the benefits of the Capon algorithm over the conventional delay and-sum approach. On heart sector images, the clutter in the heart chambers is decreased. The endocardium border is better defined. On abdominal linear array images, significant contrast and resolution enhancement are observed. PMID- 18407852 TI - Experimental validation of ultrasonic guided modes in electrical cables by optical interferometry. AB - In this work, the dispersion curves of elastic waves propagating in electrical cables and in bare copper wires are obtained theoretically and validated experimentally. The theoretical model, based on Gazis equations formulated according to the global matrix methodology, is resolved numerically. Viscoelasticity and attenuation are modeled theoretically using the Kelvin-Voigt model. Experimental tests are carried out using interferometry. There is good agreement between the simulations and the experiments despite the peculiarities of electrical cables. PMID- 18407853 TI - Blind detection of nonlinearities in multiple-echo ultrasonic signals. AB - In this paper, we propose and analyze by means of simulations the use of surrogate data algorithms for blind detection of nonlinearities in multiple-echo ultrasonic signals. We assume a blind scheme so that no information about the input (emitted ultrasonic pulse) can be used. The metrics and equations that model some nonlinear situations are carefully reviewed. Also, closed form equations of the third-order metrics from a simplified second-order Volterra kernel are derived. Computer simulations show that the surrogate data technique is a potentially powerful tool for blind detection of nonlinearities in multiple echo ultrasonic signals if adequate metrics are chosen. They also reveal interesting trade-offs among parameters that model ultrasonic systems and detection percentages. PMID- 18407854 TI - Theoretical analyses and numerical simulations of the torsional mode for two acoustic viscometers with preliminary experimental tests. AB - A rigorous analysis of the torsional modes in both a cylindrical wave guide and the associated static viscous fluid field has been conducted from the solid and the fluid wave equations and the coupled boundary conditions. As a result, two acoustic viscometer models, along with four independent equations connecting the density and the viscosity of the fluid with the attenuation and the phase velocity of the torsional wave in the wave guide, have been developed. The analysis shows that the product of the viscosity and the density of the fluid can be measured from the end reflection coefficient of the torsional wave in the wave guide and that both the viscosity and the density can be determined simultaneously from either the phase velocity or the attenuation of the torsional wave in a single cylindrical wave guide. For the simultaneous measurements of the viscosity and the density, the independent equations have to be solved numerically, for example, using Matlab (The MathWorks, Natick, MA), given either the attenuation or the phase velocity in the wave guide that is surrounded by the fluid. To demonstrate the technical feasibility, numerical simulations have been conducted to discern viscosity, phase velocity, and density, all versus attenuation, at different frequencies, and with variable dimension of a molybdenum rod, so that both the advantages and the disadvantages of the simultaneous measurements can be explored. In the end, to test the two models, preliminary experiments on two viscous standards were conducted at 23 degrees C, and good agreements have been achieved between the viscosities measured from both models and for both standards. PMID- 18407855 TI - Optimization of a new type of ultrasonic linear motor. AB - This paper presents the design, the properties, and the optimization study of a new type of ultrasonic linear motor. Numerical modeling has been carried out and simulations with software have been realized. To avoid performing a large number of simulations, sensitivity analysis has been carried out, in particular using design of experiments. The Doehlert method has been chosen in our study. The results found show that this preoptimization stage allows one to improve the deformation amplitude and to reduce the input parameter variation ranges. Finite element (FE) optimization is then carried out, and results show that the motion amplitudes can be increased compared to the initial design of the motor. Some experiments on prototypes show that the travel range of the motor has been increased while decreasing the applied voltage by a factor of 2. PMID- 18407856 TI - Adjusting the resonant frequency of a PVDF bimorph power harvester through a corrugation-shaped harvesting structure. AB - We propose a corrugated polyvinylidene fluoride (PVDF) bimorph power harvester with the harvesting structure fixed at the two edges in the corrugation direction and free at the other edges. The resonant frequency of a corrugated PVDF bimorph is readily adjusted through changing either its geometrical configuration or the span length, which can keep the harvester operating at the optimal state in environments with different ambient vibrations. The governing equations of a PVDF bimorph with a corrugation shape are derived from the transfer-matrix technique. Statistical results show that the adaptability of a harvester to the operating environment can be improved greatly by designing the harvesting structure with adjustable resonant frequency. PMID- 18407857 TI - Effect of loading a plate with different liquids on the propagation of lamb-like waves studied by laser ultrasonics. AB - Three experimental laser ultrasonic configurations--line excitation with scanning detection, grating excitation with single point detection, and grating excitation with scanning probe beam--are shown to consistently reveal the modified propagation properties of Lamb waves on a polyethylene terephthalate (PET) film that is in contact with different liquids on its two sides. Theoretical predictions concerning the physical nature of different wave modes in symmetric and asymmetric film loading configurations (i.e., their existence, velocity, damping, and polarization) are confirmed by the experimental results. PMID- 18407858 TI - 3-D electrostatic hybrid element model for SAW interdigital transducers. AB - In this work, the singular behavior of charges at corners and edges on the metallized areas in SAW transducers are investigated. In particular, it is demonstrated that a tensor product of the commonly used Tchebychev bases overestimates the singularities at corners, and, hence, it cannot be used in a proper boundary element method formulation. On the other hand, it is shown that a simple finite element method-like approach is impractical due to the enormous number of unknowns required to model the electrode's large length-to-width ratio. These considerations are then used for defining a hybrid element model, which combines Tchebychev and linear polynomials over differently meshed domains. Such an approach is shown to suitably account for charge singularities while greatly reducing the number of unknowns. Results are obtained for isotropic and anisotropic substrates for non-periodic configurations. PMID- 18407859 TI - Orthogonal frequency coded filters for use in ultra-wideband communication systems. AB - The use of ultra-short pulses, producing very wide bandwidths and low spectral power density, are the widely accepted approach for ultra-wideband (UWB) communication systems. This approach is simple and can be implemented with current digital signal processing technologies. However, surface acoustic wave (SAW) devices have the capability of producing complex signals with wide bandwidths and relatively high frequency operation. This approach, using SAW based correlators, eliminates many of the costly components that are needed in the IF block in the transmitter and receiver, and reduces many of the signal processing requirements. This work presents the development of SAW correlators using orthogonal frequency coding (OFC) for use in UWB spread spectrum communication systems. OFC and pseudonoise (PN) coding provide a means for UWB spreading of data. The use of OFC spectrally spreads a PN sequence beyond that of code division multiple access (CDMA) because of the increased bandwidth providing an improvement in processing gain. The transceiver approach is still very similar to that of a CDMA but provides greater code diversity. Experimental results of a SAW filter designed with OFC transducers are presented. The SAW correlation filter was designed using seven contiguous chip frequencies within the transducer. SAW correlators with a 29% fractional bandwidth were fabricated on lithium niobate (LiNbO3) having a center frequency of 250 MHz. A coupling-of modes (COM) model is used to predict the SAW filter response experimentally and is compared to the measured data. Good correlation between the predicted COM responses and the measured device data is obtained. Discussion of the design, analysis, and measurements are presented. The experimental matched filter results are shown for the OFC device and are compared to the ideal correlation. The results demonstrate the OFC SAW device concept for UWB communication transceivers. PMID- 18407860 TI - Modeling of bulk acoustic wave devices built on piezoelectric stack structures: impedance matrix analysis and network representation. AB - The fundamental electro-acoustic properties of a solid layer are deduced in terms of its impedance matrix (Z) and represented by a network for modeling the bulk acoustic wave devices built on piezoelectric stacked structures. A piezoelectric layer is described by a three-port equivalent network, a nonpiezoelectric layer, and a short- or open-circuit piezoelectric layer by a two-port one. Electrical input impedance of the resonator is derived in terms of the Z-matrix of both the piezoelectric layer and an external load, the unique expression applies whether the resonator is a mono- or electroded-layer or a solidly mounted resonator (SMR). The loading effects of Al-electrodes on the resonating frequencies of the piezoelectric ZnO-layer are analyzed. Transmission and reflection properties of Bragg mirrors are investigated along with the bulk radiation in SMR. As a synthesizing example, a coupled resonator filter (CRF) is analyzed using the associated two-port equivalent network and by calculating the power transmission to a 50Omega-load. The stacked crystal filter is naturally included in the model as a special case of CRF. Combining a comprehensive matrix analysis and an instructive network representation and setting the problem with a full vectorial formalism are peculiar features of the presented approach. PMID- 18407861 TI - Nonlinear dynamic analysis of traveling wave-type ultrasonic motors. AB - In this paper, nonlinear dynamic response of a traveling wave-type ultrasonic motor was investigated. In particular, understanding the transient dynamics of a bar-type ultrasonic motor, such as starting up and stopping, is of primary interest. First, the transient response of the bar-type ultrasonic motor at starting up and stopping was measured using a laser Doppler velocimeter, and its driving characteristics are discussed in detail. The motor is shown to possess amplitude-dependent nonlinearity that greatly influences the transient dynamics of the motor. Second, a dynamical model of the motor was constructed as a second order nonlinear oscillator, which represents the dynamics of the piezoelectric ceramic, stator, and rotor. The model features nonlinearities caused by the frictional interface between the stator and the rotor, and cubic nonlinearity in the dynamics of the stator. Coulomb's friction model was employed for the interface model, and a stick-slip phenomenon is considered. Lastly, it was shown that the model is capable of representing the transient dynamics of the motor accurately. The critical parameters in the model were identified from measured results, and numerical simulations were conducted using the model with the identified parameters. Good agreement between the results of measurements and numerical simulations is observed. PMID- 18407862 TI - Coupling to extension in a thickness-shear resonator due to relatively large thickness-shear deformation. AB - We show that in a plate thickness-shear mode resonator of rotated Y-cut quartz coupling to extension occurs when the shear deformation is no longer infinitesimal. A set of coupled equations is derived with which the effect of coupling to extension on the thickness-shear operating mode is examined. PMID- 18407863 TI - A comparison of time-domain solutions for the full-wave equation and the parabolic wave equation for a diagnostic ultrasound transducer. AB - A study of numerical solutions to the linear wave equation and the parabolic wave equation is presented. Finite-difference, time-domain methods are used to calculate the acoustic field emitted from a phased diagnostic ultrasound transducer in a non-attenuating medium. Results are compared to Field II, a simulation package that has been used extensively to linearly model transducers in ultrasound. The simulation of the parabolic equation can accurately predict the lateral beamplot for large f/#s, but, it exhibits 2-3 dB errors for small f/#s. It also overestimates the depth at which the focus occurs. For the considered array, it is shown that the finite-difference solution of the wave equation is accurate for a small and large f/#. The lateral beamplots and axial intensities are in excellent agreement with the Field II simulations. PMID- 18407864 TI - On the eigenvalue problem for free vibrations of a piezoelectric/piezomagnetic body. AB - We present a systematic analysis of the eigenvalue problem associated with free, small-amplitude vibrations in an elastic body possessing piezoelectric, piezomagnetic, and magnetoelectric couplings. An abstract formulation is introduced. The operators in the abstract formulation are shown to be self adjoint, from which a series of fundamental properties of resonant frequencies and vibration modes are proved concisely. A variational formulation and a perturbation analysis of the eigenvalue problem are also given, based on the abstract formulation. PMID- 18407866 TI - Effects of progesterone on benzene toxicity in rats. AB - Benzene is a frequently used industrial solvent. Its toxic manifestations could be modified by sex hormones, but mechanisms of their action are poorly understood. We have examined the influence of progesterone on lipid peroxidation (malondialdehyde), reduced glutathione (GSH), and cytochrome P450 2E1 (CYP2E1) in the liver and kidneys of female rats. Progesterone applied to benzene-treated rats inhibited the formation of reactive oxygen species (ROS), but in ovariectomised benzene-treated rats it significantly increased GSH in the liver. No improvement in CYP2E1 activity was observed in progesterone treated rats. Our results evidence that progesterone changes benzene toxicity (generation of ROS, oxidative stress). However, the probable antioxidative effect of progesterone needs to be confirmed by further studies. PMID- 18407867 TI - Risk factors for laryngeal cancer in Montenegro. AB - Laryngeal cancer is the most common head and neck cancer. There might be many risk factors for laryngeal cancer. Smoking, especially cigarette smoking and alcohol are indisputable risk factors. The authors of this paper assessed the presumed risk factors in order to identify possible aetiological agents of the disease.A hospital-based case-control study was conducted. The study group consisted of 108 histologically verified laryngeal cancer patients and 108 hospital controls matched by sex, age (+/-3 years) and place of residence. Laryngeal cancer patients and controls were interviewed during their hospital stay using a structured questionnaire. According to multiple logistic regression analysis six variables were independently related to laryngeal cancer: hard liquor consumption (Odd Ratio/OR/=2.93, Confidence Interval/CI/95% = 1.17 to 7.31), consumption more than 2 alcoholic drinks per day (OR=4.96, CI 95% = 2.04 to 12.04), cigarette smoking for more than 40 years (OR=4.32, CI 95% = 1.69 to 11.06), smoking more than 30 cigarettes per day (OR=4.24, CI 95% = 1.75 to 10.27), coffee consumption more than 5 cups per day (OR=4.52, CI 95% = 1.01 to 20.12) and carbonated beverage consumption (OR=0.38, CI 95%=0.16 to 0.92). The great majority of laryngeal cancers could be prevented by eliminating tobacco smoking and alcohol consumption. PMID- 18407868 TI - Adverse effects in workers exposed to inorganic lead. AB - This paper describes a retrospective cohort study comparing 60 workers occupationally exposed to inorganic lead and 60 matched controls. All subjects were assessed using data obtained from a specially designed Questionnaire for lead exposure and toxic effects assessment, physical examination, spirometry, ECG, and laboratory tests including blood lead level (BLL) and biomarkers of lead toxic effects. Muscle pain, droopiness, and work-related nasal symptoms were significantly more frequent in lead workers. The prevalence of lung symptoms was higher in lead workers than in controls, but not significantly (20 % vs. 6.6 %, respectively). Mean values of BLL and delta-aminolevulinic acid (ALA) were significantly higher in lead workers. The activity of delta-aminolevulinic acid dehydratase (ALAD) in lead workers was significantly lower than in controls. Abnormal of BLL, ALAD, and ALA were more frequent in lead workers, with statistical difference for BLL and ALAD. Inverse correlation was found between BLL and ALAD, and positive correlation between BLL and age, years of employment, and years of exposure. Inverse correlation was found between ALAD and age, years of employment, years of exposure, blood pressure, alcohol consumption, and years of alcohol consumption. Changes in spirometry correlated inversely with BLL. A positive correlation was found between BLL and erythrocyte count and haemoglobin concentration, whereas it was inverse for ALAD and haemoglobin concentration. A significant difference was found for BLL and ALAD, with a very high odds ratio (14.64 and 7.23, respectively) and high relative risk (4.18 and 3.08, respectively). Our data have confirmed the association between occupational lead exposure and deviation in specific biological markers of lead effect and between the role of occupational exposure in the development of adverse effects. PMID- 18407869 TI - Severe acute copper sulphate poisoning: a case report. AB - As copper sulphate pentahydrate (CSP) is a common compound used in agriculture and industry, chronic occupational exposures to CSP are well known, but acute poisoning is rare in the Western world. This case report describes acute poisoning of a 33-year-old woman who attempted suicide by ingesting an unknown amount of CSP. On admission to the hospital, she had symptoms and signs of severe hemorrhagic gastroenteritis, dehydration, renal dysfunction and methaemoglobinaemia with normal serum copper level. Therapy included early gastric lavage, fluid replacement, vasoactive drugs, furosemide, antiemetic drugs, ranitidine, and antidotes methylene blue and 2,3-dimercaptopropane-1 sulphonate (DMPS). However, the patient developed severe intravascular haemolysis, acute severe hepatic and renal failure, as well as adrenal insufficiency. After prolonged, but successful hospital treatment, including haemodialysis and IV hydrocortisone, the patient was discharged with signs of mild renal and liver impairment. Our conclusion is that in severe cases of copper poisoning early supportive measures are essential. In addition, antidotes such as methylene blue for methaemoglobinaemia and chelating agent such as DMPS improve morbidity and survival of severely poisoned victims. PMID- 18407870 TI - T-2 toxin: incidence and toxicity in poultry. AB - T-2 toxin is the most toxic type A trichothecene mycotoxin. It is the secondary metabolite of the Fusarium fungi, and is common in grain and animal feed. Toxic effects have been shown both in experimental animals and in livestock. It has been implicated in several outbreaks of human mycotoxicoses. Toxic effects in poultry include inhibition of protein, DNA, and RNA synthesis, cytotoxicity, immunomodulation, cell lesions in the digestive tract, organs and skin, neural disturbances and low performance in poultry production (decreased weight gain, egg production, and hatchability). Concentrations of T-2 toxin in feed are usually low, and its immunosuppressive effects and secondary infections often make diagnosis difficult. If at the onset of the disease, a change in diet leads to health and performance improvements in animals, this may point to mycotoxin poisoning. Regular control of grain and feed samples is a valuable preventive measure, and it is accurate only if representative samples are tested. This article reviews the incidence and toxic effects of T-2 toxin in poultry. PMID- 18407871 TI - Non-hip peripheral osteoporotic fractures: epidemiology and significance. AB - Fractures are the most serious consequence of osteoporosis. Non-vertebral and non hip fractures are seldom recognised as important, even though they account for the majority of all fractures. The most prevalent localisations are distal radius, proximal humerus, ribs, clavicle, and the pelvis. According to the results from large phase III clinical trials for placebo groups, their incidence ranges from 4.9 % to 12.0 %. Hospital morbidity data in Croatia in 2006 show that peripheral non-hip fractures ranked among the leading fifteen injuries, accounting for 23.7 % of all injuries in patients aged 60 years and above. Risk factors for non-hip and non-vertebral fractures are similar to other osteoporotic fractures, and the main are low bone mineral density and earlier fractures. Quality of life is considerably affected by these fractures, and medical costs are very high, soaring as high as 36.9% of all national medical costs in the USA. Non-vertebral non-hip fractures need more attention, which was also recognised by the European regulatory bodies that approve use of anti-osteoporotic drugs. PMID- 18407872 TI - Mycotoxic and aristolochic acid theories of the development of endemic nephropathy. AB - Despite many efforts of scientists and epidemiologists, the aetiology of endemic nephropathy (EN) is still unknown. This disease occurs in the rural population of geographically limited areas of Bulgaria, Bosnia and Herzegovina, Croatia, Romania, and Serbia, and a number of theories have been proposed about its aetiology. The mycotoxin theory has prevailed until now, based on the studies of nephrotoxic mycotoxin ochratoxin A (OTA) that revealed higher frequency of OTA positive food and blood samples in endemic than in non-endemic areas.However, a new aristolochic acid (AA) theory of EN origin has been proposed recently, due to the histological similarities in kidney lesions between patients suffering from EN and patients suffering from Chinese herbs nephropathy caused by AA. Until now it has not been unequivocally proved that the inhabitants of EN areas are exposed to higher concentration of AA than in other regions and the exposure pathways are rather uncertain. This paper presents most important studies supporting both theories, indicating also the inconsistencies of each. PMID- 18407873 TI - Supported employment in the context of psychiatric rehabilitation. PMID- 18407874 TI - Supported employment: 1998 to 2008. PMID- 18407875 TI - In favor of science. PMID- 18407876 TI - An update on randomized controlled trials of evidence-based supported employment. AB - BACKGROUND: The Individual Placement and Support (IPS) model of supported employment for clients with severe mental illness has been described as a standardization of evidence-based supported employment. Although several reviews on the literature on its effectiveness have been conducted, the completion of several new studies suggests an updated review is warranted. METHODS: We conducted a comprehensive literature search for randomized controlled trials of IPS, limiting our review to programs with high-fidelity IPS programs, locating 11 studies. We examined the following competitive employment outcomes: employment rates, days to first job, annualized weeks worked, and job tenure in longest job held during the follow-up period. FINDINGS: Across the 11 studies, the competitive employment rate was 61% for IPS compared to 23% for controls. About two-thirds of those who obtained competitive employment worked 20 hours or more per week. Among those who obtained a competitive job, IPS participants obtained their first job nearly 10 weeks earlier than did controls. Among IPS participants who obtained competitive work, duration of employment after the start of the first job averaged 24.2 weeks per year, or 47% of the 52-week year. CONCLUSIONS: The current review is consistent with earlier reviews, although the evidence for high-fidelity IPS appears to be somewhat stronger here than in reviews evaluating studies with more heterogeneity in the supported employment models examined. The number, consistency, and effect sizes of studies of evidence-based supported employment establish it as one of the most robust interventions available for persons with severe mental illness. PMID- 18407877 TI - The employment intervention demonstration program: major findings and policy implications. AB - This article summarizes the published results of the Employment Intervention Demonstration Program (EIDP), a federally-funded, multi-site study examining the effectiveness of supported employment programs for 1273 unemployed individuals with psychiatric disabilities in the U.S. Findings confirm the effectiveness of supported employment across different models, program locations, and participant populations. The study's results are discussed in the context of public policies designed to encourage return to work for those with a severe mental illness. PMID- 18407878 TI - Strategies for state-wide implementation of supported employment: the Johnson & Johnson-Dartmouth Community Mental Health Program. AB - This paper reviews key strategies for implementing evidence-based supported employment statewide. The Johnson & Johnson-Dartmouth Community Mental Health Program collaborates with 9 states and the District of Columbia to implement supported employment in a sustainable way. Technical assistance and teambased training help agencies develop high fidelity programs that result in good employment outcomes for people with severe mental illness. PMID- 18407879 TI - Fidelity of supported employment: lessons learned from the National Evidence Based Practice Project. AB - PURPOSE: To describe the implementation of supported employment in the National Evidence-Based Practices Project, examining and interpreting fidelity changes over two years of program implementation. METHOD: Nine new supported employment programs were evaluated by examining supported employment fidelity ratings at baseline and every 6 months thereafter. Site reports based on the nine case studies were used to interpret the fidelity findings. RESULTS: At two years, eight of nine sites achieved high fidelity. Most changes occurred within the first year. Twelve of the 15 supported employment fidelity items showed rapid improvement. DISCUSSION: A review of the site reports suggested four factors that facilitated movement toward high fidelity: (1) Discontinuing non-evidence-based vocational services, (2) Making rapid structural changes through administrative action, (3) Measuring key process indicators to move toward desired changes, and (4) Gradually improving integration of the employment specialists with clinical services, primarily through supervisor leadership. PMID- 18407880 TI - The mental health treatment study. AB - Security Disability Insurance (SSDI) beneficiaries with primary psychiatric impairments comprise the largest, fastest growing, and most costly population in the SSDI program. The Mental Health Treatment Study provides a comprehensive test of the hypothesis that access to evidence-based employment services and behavioral health treatments, along with insurance coverage, can enable SSDI beneficiaries with psychiatric impairments to return to competitive employment. It will also examine which beneficiaries choose to enter an employment study under such conditions. Currently in the field in 22 cities across the U.S., the MHTS aims to recruit 3,000 SSDI beneficiaries with psychiatric impairments into a randomized controlled trial. This paper describes the MHTS, its background, and its process and outcome assessments. PMID- 18407881 TI - IPS in Europe: the EQOLISE trial. AB - BACKGROUND: IPS has been demonstrated to increase return to open employment significantly in individuals with mental health problems in the US. Previous experience (e.g. with ACT) has demonstrated the sensitivity of complex community mental health interventions to local social and healthcare cultures. Europe has conditions of generally greater employment security than the US and varying (generally higher) unemployment rates and welfare benefits. Evidence of the effectiveness of IPS in these conditions, and its potential variation across them, would guide local policy and provide possible insights into its mechanism. METHODS: We conducted an RCT of IPS versus high-quality train-and-place vocational rehabilitation in six European centers with very different labor market and health and social care conditions. A sample of 312 individuals with psychotic illness was randomly allocated (50 per site). Inclusion criteria were a minimum of two years' illness duration, with at least one year of continuous unemployment and six months contact with their current mental health services. Follow-up was 18 months. The primary outcome was any open employment, and secondary outcomes included time to employment, duration of employment and hospital admission. FINDINGS: IPS was more effective than the Vocational Services for all vocational outcomes. 85 IPS patients (54.5%) worked for at least one day compared to 43 Vocational Service patients (27.6). They were significantly less likely to be rehospitalized. Local unemployment rates explained a significant amount of the variation in IPS effectiveness and both national economic growth and welfare systems influenced overall employment rates in both services. CONCLUSIONS: IPS doubles the access to work of people with psychotic illnesses, without any evidence of increased relapse. Its effectiveness is not independent of external circumstances, particularly local unemployment rates. PMID- 18407882 TI - Expert ratings on the critical ingredients of supported employment for people with severe mental illness. AB - BACKGROUND: Supported employment is identified as an evidence-based practice for people with severe mental illness, with Individual Placement and Support (IPS) emerging as the clearest articulation of this practice. Yet throughout the U.S., programs labeled as supported employment operate in a wide variety of ways, with different staffing patterns, organizational features, and services. Consensus on critical ingredients is crucial for establishing standards for evidence-based services, guiding implementation of new programs, and fostering communication in the literature. The current study examined the degree of agreement on the critical ingredients of supported employment among experts and practitioners. METHODS: The expert sample (n=19) consisted primarily of university-based researchers, while the practitioner sample (n=55) was a convenience sample generated from lists provided by state leaders in 2 Western, 2 Central, and 2 Eastern states. Using multiple sources, including published guidelines and 3 supported employment fidelity scales, we developed a 59-item survey checklist. Respondents rated items on a 7-point importance scale and indicated ideal specifications in 11 areas. FINDINGS: We found strong agreement on the critical ingredients of supported employment within the expert and practitioner groups. On most items, the two groups had similar mean ratings of importance. Experts rated 4 items reflecting evidence-based principles and 2 items related to outcome monitoring as more important than did practitioners, while practitioners rated as more important 10 items reflecting pragmatic concerns (e.g., funding, accreditation). Comparing survey responses on the 15 items from the IPS Fidelity Scale suggested general endorsement of IPS principles, except in the area of staffing. Ideal model specifications included caseload size of 16 and twice weekly supervision. Respondents also suggested critical ingredients not measured by the IPS Fidelity Scale, such as benefits counseling, supported education, a cultivation of employers. CONCLUSIONS: This survey shows a growing consensus within the psychiatric rehabilitation field on IPS principles while suggesting additional strategies endorsed by experts and practitioners. PMID- 18407883 TI - VR closure rates for two vocational models. AB - BACKGROUND: The Individual Placement and Support (IPS) model of supported employment is an evidence-based practice for individuals with psychiatric disabilities. To be financially viable, IPS programs require funding from the state-federal vocational rehabilitation (VR) system. However, some observers have questioned the compatibility of IPS and the VR system. METHOD: Using a randomized controlled trial comparing IPS to a well-established vocational program called the Diversified Placement Approach (DPA), we examined rates of VR sponsorship and successful VR closures. We also describe the establishment of an active collaboration between a psychiatric rehabilitation agency and the state VR system to facilitate rapid VR sponsorship for IPS clients. FINDINGS: Both IPS and DPA achieved a 44% rate of VR Status 26 closure when considering all clients entering the study. IPS and DPA averaged similar amount of time to achieve VR sponsorship. Time from vocational program entry to Status 26 was 51 days longer on average for IPS. CONCLUSIONS: Even though several IPS principles seem to run counter to VR practices, such as zero exclusion and rapid job search, we found IPS closure rates comparable to those for DPA, a vocational model that screens for readiness, provides prevocational preparation, and extensively uses agency-run businesses. PMID- 18407884 TI - Individual placement and support for individuals with recent-onset schizophrenia: integrating supported education and supported employment. AB - OBJECTIVE: To describe the adaptation of the Individual Placement and Support model of supported employment to individuals with a recent first episode of schizophrenia or a related psychotic disorder. METHODS AND RESULTS: Given that the vocational goals of persons with a recent onset of schizophrenia often involve completion of schooling rather than only competitive employment, the principles of Individual Placement and Support were extended to include supported education. This extension involved initial evaluation of the most appropriate goal for individual participants, having the IPS specialist working on placement either with the participant or directly with educational and employment settings (depending on permitted disclosure and individual need), and follow-along support that included work with teachers and aid in study skills and course planning as well as typical supported employment activities. Work with family members also characterized this application of IPS. A randomized controlled trial is comparing the combination of IPS and skills training with the Workplace Fundamentals Module with the combination of brokered vocational rehabilitation and broad-based social skills training. Participants in the IPS condition have returned to school, competitive work, and combined school and work with approximately equal frequency. CONCLUSIONS: IPS principles can be successfully extended to integrate supported education and supported employment within one treatment program. The distribution of return to school, work or their combination in this group of individuals with recent-onset schizophrenia supports the view that an integrated program of supported education and supported employment fits this initial period of illness. PMID- 18407885 TI - Cognitive remediation and vocational rehabilitation. AB - Persons with severe mental illness (SMI) who are striving to improve their work prospects are often hindered in work endeavors because of difficulties with cognitive skills, such as paying attention or concentrating, learning and remembering information, responding in a reasonable amount of time to environmental demands, and planning ahead and solving problems. In addition to limiting work functioning, cognitive impairments are obstacles to receiving the full benefits of vocational rehabilitation, including supported employment. Efforts to improve cognition in people with SMI, or cognitive remediation, have produced modest but consistent gains in a variety of cognitive domains. More recent efforts have focused on combining cognitive remediation with vocational rehabilitation in order to improve work functioning. Initial results from four published studies of combined cognitive remediation and vocational programs are encouraging, indicating improvements in both cognitive and work functioning. The approaches to cognitive remediation used in these studies vary considerably, as do the characteristics of participants, the vocational rehabilitation models, and the methods of combining cognitive and vocational therapies. The differences in key components of programs combining cognitive remediation and vocational rehabilitation indicate the need to replicate findings, and raise important questions about what aspects of the programs are associated with improvements in work. PMID- 18407886 TI - Factors influencing the delivery of evidence-based supported employment in England. AB - This paper assesses the extent to which the Individual Placement and Support (IPS) approach is currently adopted in England. Interviews based on the Supported Employment Fidelity Scale were conducted with staff from five of the leading providers of supported employment. One provider obtained a good IPS adherence score, three a fair score and one a non-adherence score. Constraints influencing providers' capacity to provide an IPS service related to funding, values and organizational policy. The authors discuss the implications of these constraints in relation to the recent commissioning guidance for vocational services in the UK. PMID- 18407887 TI - The future of supported employment for people with severe mental illness. AB - This paper reviews current research on innovative attempts to improve the dissemination and effectiveness of supported employment. The domains of active investigation include: (1) organization and financing of services, (2) disability policies, (3) program implementation and quality, (4) motivation, (5) job development, (6) illness-related barriers, (7) job supports, (8) career development, and (9) new populations. Work in each of these areas offers the promise of improving services and outcomes in the near future. PMID- 18407888 TI - The medium isn't the message. PMID- 18407890 TI - Scaling of xylem vessels and veins within the leaves of oak species. AB - General models of plant vascular architecture, based on scaling of pipe diameters to remove the length dependence of hydraulic resistance within the xylem, have attracted strong interest. However, these models have neglected to consider the leaf, an important hydraulic component; they assume all leaves to have similar hydraulic properties, including similar pipe diameters in the petiole. We examine the scaling of the leaf xylem in 10 temperate oak species, an important hydraulic component. The mean hydraulic diameter of petiole xylem vessels varied by 30% among the 10 oak species. Conduit diameters narrowed from the petiole to the midrib to the secondary veins, consistent with resistance minimization, but the power function scaling exponent differed from that predicted for stems. Leaf size was an organizing trait within and across species. These findings indicate that leaf vasculature needs to be included in whole-plant scaling models, for these to accurately reflect and predict whole-plant transport and its implications for performance and ecology. PMID- 18407891 TI - Sophisticated diagnostic modalities. PMID- 18407893 TI - The predictive receiver operating characteristic curve for the joint assessment of the positive and negative predictive values. AB - Binary test outcomes typically result from dichotomizing a continuous test variable, observable or latent. The effect of the threshold for test positivity on test sensitivity and specificity has been studied extensively in receiver operating characteristic (ROC) analysis. However, considerably less attention has been given to the study of the effect of the positivity threshold on the predictive value of a test. In this paper we present methods for the joint study of the positive (PPV) and negative predictive values (NPV) of diagnostic tests. We define the predictive receiver operating characteristic (PROC) curve that consists of all possible pairs of PPV and NPV as the threshold for test positivity varies. Unlike the simple trade-off between sensitivity and specificity exhibited in the ROC curve, the PROC curve displays what is often a complex interplay between PPV and NPV as the positivity threshold changes. We study the monotonicity and other geometric properties of the PROC curve and propose summary measures for the predictive performance of tests. We also formulate and discuss regression models for the estimation of the effects of covariates. PMID- 18407892 TI - Estimation of vaccine efficacy in a repeated measures study under heterogeneity of exposure or susceptibility to infection. AB - Vaccine efficacy (VE) is commonly estimated through proportional hazards modelling of the time to first infection or disease, even when the event of interest can recur. These methods can result in biased estimates when VE is heterogeneous across levels of exposure and susceptibility in subjects. These two factors are important sources of unmeasured heterogeneity, since they vary within and across areas, and often cannot be individually quantified. We propose an estimator of VE per exposure that accounts for heterogeneous susceptibility and exposure for a repeated measures study with binary recurrent outcomes. The estimator requires only information about the probability distribution of environmental exposures. Through simulation studies, we compare the properties of this estimator with proportional hazards estimation under the heterogeneity of exposure. The methods are applied to a reanalysis of a malaria vaccine trial in Brazil. PMID- 18407894 TI - Point and interval estimation of accuracies of a binary medical diagnostic test following group sequential testing. AB - When hypotheses concerning the sensitivity and specificity of a binary medical diagnostic test are simultaneously tested using a group sequential procedure, constructing point and interval estimates of the parameters is challenging because there is no unique way to order sample points in the two-dimensional space. In this paper, upon termination of a group sequential procedure, we compare the bias and mean squared errors of the maximum-likelihood and Rao Blackwell unbiased estimators of sensitivity and specificity. Confidence intervals (CIs) of the two parameters were constructed using normal approximation and Woodroofe's pivot methods based on maximum-likelihood and Rao-Blackwell unbiased estimates. The coverage probability and the expected length of CIs for the parameters were compared by simulation studies. PMID- 18407895 TI - A mathematical approach for the transactivation of hERalpha. AB - Several reports over the last few years have documented the dose-response curve for steroid hormone induction of gene transcription as a modulated property of a given receptor-agonist complex that varies with the changing concentration of a variety of factors including: homologous receptor, co-activators, co-repressors and selected co-factors. In each report, the dose-response curves are sigmoidal and show an excellent fit with the curve generated by Michaelis-Menten kinetics. In addition, even the overall function of human oestrogen receptors (hERs) can show a similar graph for the determination of sex versus oestrogen compounds in reptiles. Thus, the kinetic properties of the simple bimolecular reaction of A+B- >C appear, surprisingly, to be sufficient to describe the dose-response curve of the multi-step process of steroid-regulated gene induction that involves several molecules. Any advance in explaining why the dose-response curve for steroid regulated gene expression is sigmoidal would assist in understanding what parameters are key factors of the dose-response curve and can benefit in the design of new oestrogenic substances. We have constructed and analysed a multi step model of hER-induced gene transcription that explains the multiple forms of a simple dose-response curve in an in vitro transcription system. PMID- 18407896 TI - Validation of image segmentation by estimating rater bias and variance. AB - The accuracy and precision of segmentations of medical images has been difficult to quantify in the absence of a 'ground truth' or reference standard segmentation for clinical data. Although physical or digital phantoms can help by providing a reference standard, they do not allow the reproduction of the full range of imaging and anatomical characteristics observed in clinical data. An alternative assessment approach is to compare with segmentations generated by domain experts. Segmentations may be generated by raters who are trained experts or by automated image analysis algorithms. Typically, these segmentations differ due to intra rater and inter-rater variability. The most appropriate way to compare such segmentations has been unclear. We present here a new algorithm to enable the estimation of performance characteristics, and a true labelling, from observations of segmentations of imaging data where segmentation labels may be ordered or continuous measures. This approach may be used with, among others, surface, distance transform or level-set representations of segmentations, and can be used to assess whether or not a rater consistently overestimates or underestimates the position of a boundary. PMID- 18407897 TI - Multiple imputation inference for multivariate multilevel continuous data with ignorable non-response. AB - Methods specifically targeting missing values in a wide spectrum of statistical analyses are now part of serious statistical thinking due to many advances in computational statistics and increased awareness among sophisticated consumers of statistics. Despite many advances in both theory and applied methods for missing data, missing-data methods in multilevel applications lack equal development. In this paper, I consider a popular inferential tool via multiple imputation in multilevel applications with missing values. I specifically consider missing values occurring arbitrarily at any level of observational units. I use Bayesian arguments for drawing multiple imputations from the underlying (posterior) predictive distribution of missing data. Multivariate extensions of well-known mixed-effects models form the basis for simulating the posterior predictive distribution, hence creating the multiple imputations. The discussion of these topics is demonstrated in an application assessing correlates to unmet need for mental health care among children with special health care needs. PMID- 18407898 TI - Improving interval estimation of binomial proportions. AB - In this paper, we propose one new confidence interval for the binomial proportion; our interval is based on the Edgeworth expansion of a logit transformation of the sample proportion. We provide theoretical justification for the proposed interval and also compare the finite-sample performance of the proposed interval with the three best existing intervals-the Wilson interval, the Agresti-Coull interval and the Jeffreys interval-in terms of their coverage probabilities and expected lengths. We illustrate the proposed method in two real clinical studies. PMID- 18407899 TI - A Bayesian precision profile for measuring the quality of immunoassay experiments. AB - A precision profile, the relationship between the concentration of a substance and its measured precision, is a convenient way of conveying the ability of an immunoassay to accurately measure the concentration of a substance in blood serum. A precision profile is characterized by the definition of precision. Historically, precision has been evaluated as the standard error of an estimator of the concentration in a sample conditional on the true concentration. In this paper, Bayesian predictive inference is used to develop a new measure of precision based on the accuracy with which an assay could infer the concentration in a hypothetical new sample. This leads to a natural procedure for evaluating a precision profile that avoids using approximations such as those inherent in traditional methods. PMID- 18407900 TI - A method of assessing the sensitivity of the Cochran-Mantel-Haenszel test to an unobserved confounder. AB - Observational studies, including the case-control design frequently used in epidemiology, are subject to a number of biases and possible confounding factors. Failure to adjust with them may lead to an erroneous conclusion about the existence of a causal relationship between exposure and disease. The Cochran Mantel-Haenszel (CMH) test is widely used to measure the strength of the association between an exposure and disease or response, after stratifying on the observed covariates. Thus, observed confounders are accounted for in the analysis. In practice, there may be causal variables that are unknown or difficult to obtain. Hence, they are not incorporated into the analysis. Sensitivity analysis enables investigators to assess the robustness of the findings. A method for assessing the sensitivity of the CMH test to an omitted confounder is presented here. The technique is illustrated by re-examining two datasets: one concerns the effect of maternal hypertension as a risk factor for low birth weight infants and the other focuses on the risk of allopurinol on having a rash. The computer code performing the sensitivity analysis is provided in appendix A. PMID- 18407901 TI - Bayesian assessment of overtriage and undertriage at a level I trauma centre. AB - We analysed the trauma triage system at a specific level I trauma centre to assess rates of over- and undertriage and to support recommendations for system improvements. The triage process is designed to estimate the severity of patient injury and allocate resources accordingly, with potential errors of overestimation (overtriage) consuming excess resources and underestimation (undertriage) potentially leading to medical errors.We first modelled the overall trauma system using risk analysis methods to understand interdependencies among the actions of the participants. We interviewed six experienced trauma surgeons to obtain their expert opinion of the over- and undertriage rates occurring in the trauma centre. We then assessed actual over- and undertriage rates in a random sample of 86 trauma cases collected over a six-week period at the same centre. We employed Bayesian analysis to quantitatively combine the data with the prior probabilities derived from expert opinion in order to obtain posterior distributions. The results were estimates of overtriage and undertriage in 16.1 and 4.9% of patients, respectively. This Bayesian approach, which provides a quantitative assessment of the error rates using both case data and expert opinion, provides a rational means of obtaining a best estimate of the system's performance. The overall approach that we describe in this paper can be employed more widely to analyse complex health care delivery systems, with the objective of reduced errors, patient risk and excess costs. PMID- 18407902 TI - Two-stage procedures for selecting the best diagnostic biomarkers. AB - Considered in the paper is the problem of selecting a diagnostic biomarker that has the highest classification rate among several candidate markers with dichotomous outcomes. The probability of correct selection depends on a number of nuisance parameters from the joint distribution of the biomarkers and thus can be substantially affected if these nuisance parameters are misspecified. A two-stage procedure is proposed to compute the needed sample size that achieves the desired level of correct selection, as so confirmed by simulation results. PMID- 18407903 TI - Nutrition support in the critically ill: a physician survey. AB - BACKGROUND: Current clinical practice guidelines delineate optimal nutrition management in the intensive care unit (ICU) patient. In light of these existing data, the authors identify current physician perceptions of nutrition in critical illness, preferences relating to initiation of feeding, and management practices specific to nutrition after initiation of feeding in the ICU patient. METHODS: The authors electronically distributed a 12-question survey to attending physicians, fellows, and residents who routinely admit patients to medical and surgical ICUs. RESULTS: On a scale ranging from 1 to 5 (1 = low, 5 = high), the attending physician's mean rating for importance of nutrition in the ICU was 4.60, the rating for comfort level with the nutrition support at the authors' institution was 3.70, and the rating for the physician's own understanding of nutrition support in critically ill patients was 3.33. Attending physicians, fellows, and residents reported waiting an average of 2.43, 1.79, and 2.63 days, respectively, before addressing nutrition status in an ICU patient. Fifty-two percent of attending physicians chose parenteral nutrition as the preferred route of nutrition support in a patient with necrotizing pancreatitis. If a patient experiences enteral feeding intolerance, physicians most commonly would stop tube feeds. There was no significant difference in responses to any of the survey questions between attending physicians, fellows, and residents. CONCLUSIONS: This study demonstrates a substantial discordance in physician perceptions and practice patterns regarding initiation and management of nutrition in ICU patients, indicating an urgent need for nutrition-related education at all levels of training. PMID- 18407904 TI - Effects of oral nutritional supplements in normally nourished or mildly undernourished geriatric patients after surgery for hip fracture: a randomized clinical trial. AB - BACKGROUND: Oral nutritional supplements have been recommended after orthopedic surgery in geriatric patients to reduce postoperative complications. However, tolerability of supplements could be a limitation, and their universal use is not supported by the heterogeneity of previous studies, especially in patients without malnutrition. METHODS: This study is a randomized, controlled, open, parallel, 3-arm clinical trial comparing supplementation with protein powder dissolved in liquids to aim at 36 g of protein per day, energy and protein supplements to aim at 37.6 g of protein and 500 kcal per day, or no intervention in normally nourished or mildly undernourished patients. Outcomes were serum albumin, prealbumin, retinol-binding globulin, and body mass index, among others. Postoperative complications were also recorded. RESULTS: Ninety patients aged 83.8 +/- 6.6 years were included. The mean ingested amount of supplements was 41.1% +/- 20.6% in the protein powder supplement group and 51.4% +/- 13.2% in the energy protein supplement group (t = 2.278, P = .027). Postoperative supplements had no effect on the nutrition status during in-hospital follow-up, as assessed by serum albumin (P = .251), prealbumin (P = .530), retinol-binding globulin (P = .552), or body mass index (P = .582). Multivariate analysis showed that length of hospital stay with an established complication until its resolution (beta = .230, P = .031), total hospital stay (beta = .450, P < .001), baseline body mass index (beta = .204, P = .045), and total daily ingested proteins per body weight (beta = .252, P = .018) were predictive variables on the change in serum albumin (R2 = 0.409, F = 11.246, P < .001). CONCLUSIONS: Oral nutritional supplements in normally nourished or only mildly undernourished geriatric patients with hip fracture submitted to surgery may be of interest for patients with postoperative complications and long hospital stays. PMID- 18407905 TI - Trace element loss in urine and effluent following traumatic injury. AB - BACKGROUND: Few data are available to establish recommendations for trace element supplementation during critical illness. This study quantified the loss of several elements and assessed the adequacy of manganese and selenium in parenteral nutrition (PN). METHODS: Men with traumatic injuries were grouped by renal status: adequate (POLY; n = 6), acute failure with continuous venovenous hemofiltration (CVVH; n = 2), or continuous venovenous hemodiafiltration (CVVHD; n = 4). PN supplied 300 microg/d manganese and 60 microg/d selenium. Urine and effluent (from artificial kidneys) were collected for 3 days and analyzed for boron, manganese, nickel, and silicon using inductively coupled plasma atomic emission spectrometry, and for selenium using atomic absorption spectrometry. RESULTS: POLY manganese and selenium excretion averaged (standard deviation [SD]) 7.9 (3.3) microg/d and 103.5 (22.4) microg/d, respectively. All elements except selenium were detected in dialysate (prior to use). CVVHD effluent contained 3.5 and 7.3 times more manganese and nickel than CVVH ultrafiltrate, respectively. Loss of manganese averaged 2.6%, 21%, and 73% of PN amounts for POLY, CVVH, and CVVHD groups, respectively. DISCUSSION: Minimal loss of manganese compared with the amount in PN suggests that excessive amounts are retained. POLY patients excreted more selenium than was in PN, indicating negative balance. POLY losses of boron and silicon were less than that published for healthy adults, reflecting less than typical intake, whereas loss during CVVH was in the normal reference range, possibly because of added intake from boron contamination of replacement fluids. All patients lost more nickel than amounts published for healthy adults. CONCLUSIONS: Current guidelines of 60-100 microg/d of parenteral manganese may be excessive for trauma patients. The uptake of manganese and nickel from contaminants in CVVHD dialysate should be investigated. PMID- 18407906 TI - Iatrogenic malnutrition in neonatal intensive care units: urgent need to modify practice. AB - BACKGROUND: Extrauterine growth retardation is a major clinical problem in very low-birth-weight infants. Parenteral nutrition (PN) serves to achieve rapid maximal nutrition in early postnatal life. There is a lack of uniformity with regard to neonatal PN practice. The objective of this study is to ascertain current practice regarding neonatal PN prescription in the early postnatal period in the United Kingdom. METHODS: A study questionnaire was e-mailed to neonatal pharmacists serving level 3 and major level 2 units in the United Kingdom between October 2005 and March 2006. Static numerical information regarding glucose, amino acids, and lipid prescription during the first 10 days of life was collected and compared with current recommendations. RESULTS: Fifty-two (81%) units responded to the questionnaire; 4 units were excluded for incomplete data. Twenty-six units (54%) initiated PN on day 1. Full PN was achieved by the median age of 6 days. Twelve units (25%) achieved full PN only by day 7 or later. Maximum median amino acids were 2.9 g/kg/d. Only 13 units (27%) prescribed >/=3 g/kg/d, and 2 prescribed more than 3.5 g/kg/d. Nineteen units (39%) initiated lipids on day 1. Eleven units (23%) delayed lipids until day 3, and 2 units delayed lipids until day 4. In comparison to the recommended intake of calories and amino acids, the current median prescription would result in a cumulative deficit over the first 10 days of 420 kcal/kg and 11.9 g/kg, respectively. CONCLUSIONS: Our study suggests diverse practice with regard to neonatal PN prescription in the United Kingdom. Current neonatal PN practice entails a significant calorie and protein deficit during early postnatal life and warrants further review. PMID- 18407907 TI - Field testing of the 2006 World Health Organization growth charts from birth to 2 years: assessment of hospital undernutrition and overnutrition rates and the usefulness of BMI. AB - BACKGROUND: The World Health Organization (WHO) recently released a growth standard, a first attempt at describing how children should grow in an ideal environment. These charts introduce body mass index (BMI)-for-age percentiles for children younger than 2 years. Adopting the WHO standard may affect the number of children screened to require follow-up; hence, field testing needs to be completed in a tertiary care center where the incidence of suboptimal nutrition is high. The objectives of this study were to quantify differences between the new WHO and 2000 Centers for Disease Control and Prevention (CDC) growth charts for children younger than 2 years. The interchangeability of the WHO weight-for length and WHO BMI percentiles was also assessed. METHODS: Percentile scores were computed for children younger than 2 years (n = 547) admitted to a pediatric tertiary health care center in Toronto, Canada. RESULTS: The WHO standard identified more children younger than 2 years as at risk of overweight/obesity compared with the CDC reference (21.0% vs 16.6%, >or=85th weight-for-length percentile) and fewer children as wasted (18.6% vs 23.0%, <5th weight-for-length percentile). The WHO BMI-for-age and WHO weight-for-length percentiles were highly correlated (r2 = 0.83) but not interchangeable. For approximately 9% of all children, and approximately 16% of those aged 25 percentile points. CONCLUSIONS: These data describe for the first time the magnitude of differences in the number of children screened as undernourished (4.4% decrease) or overnourished (4.4% increase) with adoption of the WHO standard in a tertiary care setting. Furthermore, the WHO's BMI-for-age and weight-for-length percentiles for children younger than 2 years are correlated but are not interchangeable. PMID- 18407908 TI - Sesame oil attenuates hepatic lipid peroxidation by inhibiting nitric oxide and superoxide anion generation in septic rats. AB - BACKGROUND: Sepsis is a major cause of mortality in the intensive care unit. Oxidative stress plays an important role in the pathogenesis of organ failure during sepsis. Sesame oil decreases circulating oxygen free radicals in septic rats; however, its effect on hepatic oxidative status is unknown. The authors examined the effect of sesame oil on hepatic lipid peroxidation in septic rats. METHODS: Hepatic injury was induced using cecal ligation and puncture (CLP). Rats were divided into 4 groups: sham, rats given a sham operation without CLP; SO, rats given sesame oil alone; CLP, rats given saline and then CLP; and CS, rats given sesame oil and then CLP. All rats were first given a 1-week daily oral supplement of sesame oil or saline (4 mL/kg/d) and then CLP or a sham operation. The authors assessed hepatic oxidative stress by determining hepatic lipid peroxidation, hydroxyl radical, superoxide anion, and nitric oxide levels 12 hours after CLP. They also assessed xanthine oxidase activity and nitric oxide synthase expression. RESULTS: Hepatic lipid peroxidation (P < .0001), hydroxyl radical (P < .05), superoxide anion (P < .05), and nitrite (P < .05) levels were significantly lower in sesame oil-treated septic rats. Furthermore, sesame oil significantly reduced xanthine oxidase activity (P < .01) and inducible nitric oxide synthase expression (P < .005) in septic rats. CONCLUSIONS: Sesame oil might attenuate hepatic lipid peroxidation by inhibiting superoxide anion and nitric oxide, at least partially, in experimental septic rats. PMID- 18407909 TI - Covert assessment of concurrent and construct validity of a chart to characterize fecal output and diarrhea in patients receiving enteral nutrition. AB - BACKGROUND: An accurate and convenient method for characterizing fecal output and a consistent threshold for classifying diarrhea in patients receiving enteral nutrition are required. The aim of this study is to covertly assess the construct and concurrent validity of a chart for characterizing fecal output and classifying diarrhea in patients receiving enteral nutrition. METHODS: The chart was used to monitor fecal output in patients receiving enteral nutrition for a total of 280 patient days. Nurses characterized 291 fecal samples, of which 84 underwent measurement of fecal water using lyophilization and 60 underwent Clostridium difficile enterotoxin analysis using enzyme-linked immunosorbent assay. Construct and concurrent validity was assessed covertly to measure the true performance of the chart in a real-life clinical and research context. RESULTS: Use of the chart demonstrated higher fecal frequency (P 94% and for isoniazid, >92%. The three studies that applied NRA directly on sputum samples had a sensitivity and specificity that ranged between 88% and 100%. The SROC curve had an area of >0.99 for both drugs. CONCLUSIONS: There is evidence that NRA is highly sensitive and specific for the rapid detection of rifampicin and isoniazid resistance in culture isolates. More evidence is required for the NRA applied directly on sputum samples, but preliminary results appear promising and show a good sensitivity and specificity. Additional studies are required in countries with a high prevalence of MDR-TB and also cost-effectiveness analysis in order to obtain a complete picture on the utility of this method for rapid drug resistance detection in tuberculosis. PMID- 18407919 TI - Novel mutations in LHX3 are associated with hypopituitarism and sensorineural hearing loss. AB - Homozygous loss-of-function mutations in the transcription factor LHX3 have been associated with hypopituitarism with structural anterior pituitary defects and cervical abnormalities with or without restricted neck rotation. We report two novel recessive mutations in LHX3 in four patients from two unrelated pedigrees. Clinical evaluation revealed that all four patients exhibit varying degrees of bilateral sensorineural hearing loss, which has not been previously reported in association with LHX3 mutations, in addition to hypopituitarism including adrenocorticotropic hormone deficiency and an unusual skin and skeletal phenotype in one family. Furthermore, re-evaluation of three patients previously described with LHX3 mutations showed they also exhibit varying degrees of bilateral sensorineural hearing loss. We have investigated a possible role for LHX3 in inner ear development in humans using in situ hybridization of human embryonic and fetal tissue. LHX3 is expressed in defined regions of the sensory epithelium of the developing inner ear in a pattern overlapping that of SOX2, which precedes the onset of LHX3 expression and is known to be required for inner ear and pituitary development in both mice and humans. Moreover, we show that SOX2 is capable of binding to and activating transcription of the LHX3 proximal promoter in vitro. This study therefore extends the phenotypic spectrum associated with LHX3 mutations to encompass variable sensorineural hearing loss and suggests a possible interaction between LHX3 and SOX2 likely to be important for development of both the inner ear and the anterior pituitary in human embryonic development. PMID- 18407920 TI - De novo CoA biosynthesis is required to maintain DNA integrity during development of the Drosophila nervous system. AB - In a forward genetic screen in Drosophila melanogaster, aimed to identify genes required for normal locomotor function, we isolated dPPCS (the second enzyme of the Coenzyme A biosynthesis pathway). The entire Drosophila CoA synthesis route was dissected, annotated and additional CoA mutants were obtained (dPANK/fumble) or generated (dPPAT-DPCK). Drosophila CoA mutants suffer from neurodegeneration, altered lipid homeostasis and the larval brains display increased apoptosis. Also, de novo CoA biosynthesis is required to maintain DNA integrity during the development of the central nervous system. In humans, mutations in the PANK2 gene, the first enzyme in the CoA synthesis route, are associated with pantothenate kinase-associated neurodegeneration. Currently, the pathogenesis of this neurodegenerative disease is poorly understood. We provide the first comprehensive analysis of the physiological implications of mutations in the entire CoA biosynthesis route in an animal model system. Surprisingly, our findings reveal a major role of this conserved pathway in maintaining DNA and cellular integrity, explaining how impaired CoA synthesis during CNS development can elicit a neurodegenerative phenotype. PMID- 18407921 TI - Towards patterns tree of gene coexpression in eukaryotic species. AB - MOTIVATION: Cellular pathways behave coordinated regulation activity, and some reported works also have affirmed that genes in the same pathway have similar expression pattern. However, the complexity of biological systems regulation actually causes expression relationships between genes to display multiple patterns, such as linear, non-linear, local, global, linear with time-delayed, non-linear with time-delayed, monotonic and non-monotonic, which should be the explicit representation of cellular inner regulation mechanism in mRNA level. To investigate the relationship between different patterns, our work aims to systematically reveal gene-expression relationship patterns in cellular pathways and to check for the existence of dominating gene-expression pattern. By a large scale analysis of genes expression in three eukaryotic species, Saccharomyces cerevisiae, Caenorhabditis elegans and Human, we constructed gene coexpression patterns tree to systematically and hierarchically illustrate the different patterns and their interrelations. RESULTS: The results show that the linear is the dominating expression pattern in the same pathway. The time-shifted pattern is another important relationship pattern. Many genes from the different pathway also present coexpression patterns. The non-linear, non-monotonic and time delayed relationship patterns reflect the remote interactions between the genes in cellular processes. Gene coexpression phenomena in the same pathways are diverse in different species. Genes in S.cerevisiae and C.elegans present strong coexpression relationships, especially in C.elegans, coexpression is more universal and stronger due to its special array of genes. However in Human, gene coexpression is not apparent and the human genome involves more complicated functional relationships. In conclusion, different patterns corresponding to different coordinating behaviors coexist. The patterns trees of different species give us comprehensive insight and understanding of genes expression activity in the cellular society. PMID- 18407922 TI - Divisive Correlation Clustering Algorithm (DCCA) for grouping of genes: detecting varying patterns in expression profiles. AB - MOTIVATION: Cluster analysis (of gene-expression data) is a useful tool for identifying biologically relevant groups of genes that show similar expression patterns under multiple experimental conditions. Various methods have been proposed for clustering gene-expression data. However most of these algorithms have several shortcomings for gene-expression data clustering. In the present article, we focus on several shortcomings of conventional clustering algorithms and propose a new one that is able to produce better clustering solution than that produced by some others. RESULTS: We present the Divisive Correlation Clustering Algorithm (DCCA) that is suitable for finding a group of genes having similar pattern of variation in their expression values. To detect clusters with high correlation and biological significance, we use the correlation clustering concept introduced by Bansal et al. Our proposed algorithm DCCA produces a clustering solution without taking number of clusters to be created as an input. DCCA uses the correlation matrix in such a way that all genes in a cluster have highest average correlation with genes in that cluster. To test the performance of the DCCA, we have applied DCCA and some well-known conventional methods to an artificial dataset, and nine gene-expression datasets, and compared the performance of the algorithms. The clustering results of the DCCA are found to be more significantly relevant to the biological annotations than those of the other methods. All these facts show the superiority of the DCCA over some others for the clustering of gene-expression data. AVAILABILITY: The software has been developed using C and Visual Basic languages, and can be executed on the Microsoft Windows platforms. The software may be downloaded as a zip file from http://www.isical.ac.in/~rajat. Then it needs to be installed. Two word files (included in the zip file) need to be consulted before installation and execution of the software. PMID- 18407923 TI - Influences on child eating and weight development from a behavioral genetics perspective. AB - Childhood obesity is a strong risk factor for associated comorbidities such as type 2 diabetes, cardiovascular disease, and orthopedic abnormalities in youth and its increasing incidents thus represents a major public health concern. The following review provides evidence for a familial association between parental and child weight status, eating behaviors, and food preferences. It further draws the link between environmental influences, such as parent feeding practices, and the development of child eating behaviors and thereby elucidates how genetic and nongenetic influences can contribute to the familial transmission of obesity. We use eating in the absence of hunger, an eating trait which refers to children's susceptibility to eating in response to the presence of palatable foods in the absence of hunger, as an example to illustrate these associations. The review concludes with an outlook on possibilities for future research efforts in the field. PMID- 18407924 TI - Radio role models for the prevention of mother-to-child transmission of HIV and HIV testing among pregnant women in Botswana. AB - Although Botswana supports a program for the prevention of mother-to-child transmission of HIV (PMTCT), many women initially did not take advantage of the program. Using data from a 2003 survey of 504 pregnant and post-partum women, we assessed associations between exposure to a long-running radio serial drama that encourages use of the PMTCT program and HIV testing during pregnancy. Controlling for demographic, pregnancy and other variables, women who spontaneously named a PMTCT character in the serial drama as their favorite character were nearly twice as likely to test for HIV during pregnancy as those who did not. Additionally, multiparity, knowing a pregnant woman taking AZT, having a partner who tested, higher education and PMTCT knowledge were associated with HIV testing during pregnancy. Identification with characters in the radio serial drama is associated with testing during pregnancy. Coupled with other supporting elements, serial dramas could contribute to HIV prevention, treatment and care initiatives. PMID- 18407925 TI - Building capacity for AIDS NGOs in southern Africa: evaluation of a pilot initiative. AB - In this paper, we present the evaluation results of an AIDS non-governmental organization (NGO) capacity building 20-month pilot initiative in five countries in southern Africa called the NGO Institute. A five-person international team conducted a 2 week evaluation of the pilot in 2004 to assess the strength of the model, designed and funded by Bristol-Myers Squibb Foundation. The NGO Institute functioned through a separate consortium in each country. Results of the pilot indicate variations in adaptation and implementation of the model in each of the five countries. Each consortium took considerable time to develop its own governance and management systems. There were examples of strengthened NGO capacity in each country although it was too soon to establish overall impact. The strengths and weaknesses of this NGO capacity building model are presented along with the implications for other funding agencies and NGOs. PMID- 18407926 TI - Novel, Web-based, information-exploration approach for improving operating room logistics and system processes. AB - Routine clinical information systems now have the ability to gather large amounts of data that surgical managers can access to create a seamless and proactive approach to streamlining operations and minimizing delays. The challenge lies in aggregating and displaying these data in an easily accessible format that provides useful, timely information on current operations. A Web-based, graphical dashboard is described in this study, which can be used to interpret clinical operational data, allow managers to see trends in data, and help identify inefficiencies that were not apparent with more traditional, paper-based approaches. The dashboard provides a visual decision support tool that assists managers in pinpointing areas for continuous quality improvement. The limitations of paper-based techniques, the development of the automated display system, and key performance indicators in analyzing aggregate delays, time, specialties, and teamwork are reviewed. Strengths, weaknesses, opportunities, and threats associated with implementing such a program in the perioperative environment are summarized. PMID- 18407927 TI - Use of laparoscopy in evaluation and treatment of penetrating and blunt abdominal injuries. AB - Use of laparoscopy in penetrating trauma has been well established; however, its application in blunt trauma is evolving. The authors hypothesized that laparoscopy is safe and feasible as a diagnostic and therapeutic modality in both the patients with penetrating and blunt trauma. Trauma registry data and medical records of consecutive patients who underwent laparoscopy for abdominal trauma were reviewed. Over a 4-year period, 43 patients (18 blunt trauma / 25 penetrating trauma) underwent a diagnostic laparoscopy. Conversion to laparotomy occurred in 9 (50%) blunt trauma and 9 (36%) penetrating trauma patients. Diagnostic laparoscopy was negative in 33% of blunt trauma and 52% of penetrating trauma patients. Sensitivity/specificity of laparoscopy in patients with blunt and penetrating trauma was 92%/100% and 90%/100%, respectively. Overall, laparotomy was avoided in 25 (58%) patients. Use of laparoscopy in selected patients with blunt and penetrating abdominal trauma is safe, minimizes nontherapeutic laparotomies, and allows for minimal invasive management of selected intra-abdominal injuries. PMID- 18407928 TI - Innovative use of quality-of-life data: correlating physiologic parameters with patient-centered symptoms-- the example of anemia on the vitality of surgical oncology patients. AB - BACKGROUND: It has been difficult to correlate quality of life with physiologic parameters. This may be because of the multitude of factors that lead to a symptom. An example of a cause of fatigue, lassitude, and lack of vitality is anemia. This study explores whether a generic quality-of-life instrument's measure of vitality is correlated with anemia. METHODS: Surgical oncology patients were asked to complete the SF-36. One of the domains is vitality (VT), which is a measure of fatigue (best possible score 100, worst possible score 0). Hemoglobin (Hb, gm/dL) and hematocrit (Hct, %) levels from the same period were recorded. RESULTS: A total of 319 patients were assessed, 114 postoperative patients with no evidence of disease (NED) and 205 patients with active disease. There were no differences in Hb or Hct levels, but VT was slightly higher in NED patients. Linear regression analysis demonstrated that for overall VT, VT in patients with active malignant disease, and VT in NED patients the regression slopes were statistically significantly different from 0, although the regression coefficients (r) were all less than .5. CONCLUSIONS: There are direct correlations between Hb and Hct levels and the VT domain of the SF-36. This correlation was stronger in NED patients. The low r(2) values reflect that anemia is one of many factors affecting VT. PMID- 18407929 TI - Ultrasound-guided breast biopsy curriculum for surgical residents. AB - Ultrasound-guided breast biopsy has emerged as a common method for lesion diagnosis. This study sought to instruct and measure surgical residents' performance in ultrasound-guided breast biopsy and evaluate their thoughts regarding it. Thirteen (n = 13) senior residents completed a written pretest or questionnaire and 2.5 hours of simulated breast core and vacuum needle biopsies. Residents then completed the same written exam, and their biopsy performance was rated. There was 13% overall improvement of written test scores, and 73% resident improved comfort levels with performing biopsies. Successfully performed core biopsies and vacuum biopsies were 86% and 83%, respectively. All residents reported that instruction in ultrasound-guided breast biopsy is very important and should be mandatory in residency training programs. With concentrated instruction, residents are able to learn ultrasound-guided breast biopsy with improvement in objective measures and self-confidence levels. Resident feedback was positive and emphasized the importance of this training in surgical residency curriculums. PMID- 18407930 TI - Enhanced robotic surgical training using augmented visual feedback. AB - The goal of this study was to enhance robotic surgical training via real-time augmented visual feedback. Thirty novices (medical students) were divided into 5 feedback groups (speed, relative phase, grip force, video, and control) and trained during 1 session in 3 inanimate surgical tasks with the da Vinci Surgical System. Task completion time, distance traveled, speed, curvature, relative phase, and grip force were measured immediately before and after training and during a retention test 2 weeks after training. All performance measures except relative phase improved after training and were retained after 2 weeks. Feedback specific effects showed that the speed group was faster than other groups after training, and the grip force group applied less grip force. This study showed that the real-time augmented feedback during training can enhance the surgical performance and can potentially be beneficial for both training and surgery. PMID- 18407931 TI - In-training sounding board: do the right thing. PMID- 18407932 TI - Cancer control activities in the Republic of Korea. AB - South Korea has a population of 47.3 million. The whole population is covered by a mandatory social insurance system (the National Health Insurance Program) that is financed through the contributions paid by the insured and their employers. Cancer has been the leading cause of death in Korea since 1983. About 130 000 people develop cancer annually with 66 000 deaths in 2006. Cancer patients' 5 year survival rates between 1998 and 2002 were 37.8 and 57.0% for men and women, respectively. The five leading primary cancer sites were stomach, lung, liver, colon and rectum, and bladder among males, whereas the most common cancers were stomach, breast, colon and rectum, uterine cervix and lung among females. With the rapidly aging population, reducing cancer burden at the national level has become one of the major political issues in Korea. The government formulated its first 10-year plan for cancer control in 1996. In 2000, the National Cancer Center was created and the Cancer Control Division was set up within the Ministry of Health and Welfare. The Cancer Control Act was legislated in 2003. Korea's major national cancer control programs are anti-smoking campaigns, hepatitis B virus vaccination, cancer registration and networking, promotion of R&D activities for cancer control, education and training for cancer control and prevention, operation of the national cancer information center, operation of the mass screening program for five common cancers, management of cancer patients at home, financial support for cancer patients and designation of regional cancer centers. PMID- 18407933 TI - Capecitabine monotherapy for recurrent and metastatic nasopharyngeal cancer. AB - BACKGROUND: Capecitabine monotherapy had activity in recurrent/metastatic nasopharyngeal carcinoma (NPC) as demonstrated previously in a small pilot study. We conducted a retrospective review of patients who received capecitabine for recurrent and metastatic NPC to further evaluate its clinical benefits. METHODS: Forty-nine patients with recurrent and metastatic NPC received capecitabine at a dose of 1-1.25 G/m(2) twice daily for 14 days in 3-week cycles. Disease sites were locoregional in 29%, distant in 45% and locoregional plus distant in 26%. All except one had prior platinum-based chemotherapy for relapse or as adjunctive treatment. Median follow-up was 10 months (range: 3-41). RESULTS: Treatment was generally well tolerated. Hand-foot syndrome was common and occurred in 86% (25% Grade 3). Grade 3 hematological toxicity occurred in 6%. Partial response rate was 31% (95% CI: 18%, 44%) and complete response rate was 6% (95% CI: 0%, 13%), for an overall response rate of 37% (95% CI: 23%, 50%). Median time-to progression was 5 months and median survival was 14 months. One- and two-year survival rates were 54 and 26%, respectively. Significantly better survival was observed in patients treated for locoregional recurrence and those with severe hand-foot syndrome. CONCLUSIONS: Capecitabine has single agent activity in NPC and severe hand-foot syndrome predicts favorable outcome. Based on our experience, capecitabine monotherapy should be considered in patients with recurrent/metastatic NPC. PMID- 18407935 TI - Three cases of sub-scalp tumor presenting with protrusion of the head. PMID- 18407934 TI - Clinicopathological and prognostic relevance of uptake level using 18F fluorodeoxyglucose positron emission tomography/computed tomography fusion imaging (18F-FDG PET/CT) in primary breast cancer. AB - OBJECTIVE: Using integrated 18F-fluorodeoxyglucose positron emission tomography/computed tomography fusion imaging (18F-FDG PET/CT), the clinical significance of 18F-FDG uptake was evaluated in patients with primary breast cancer. METHODS: Clinicopathological correlation with the level of maximum standardized uptake values (SUV) 60 min obtained from preoperative 18F-FDG PET/CT were examined in 152 patients with primary breast cancer. The prognostic impact of the level of SUV was explored using simulated prognosis derived from computed program Adjuvant! in 136 (89%) patients with invasive ductal carcinoma (IDC). RESULTS: High SUV level was significantly correlated with tumor invasive size (< or = 2 cm) (P < 0.0001), higher score of nuclear grade (P < 0.0001), nuclear atypia (P < 0.0001) and mitosis counts (P < 0.0001), negative hormone receptor status (P = 0.001), high score of c-erbB-2 expression (P = 0.006), lymph node metastasis (P = 0.002), and IDC in comparison with invasive lobular carcinoma (P = 0.004). Multivariate analyses showed tumor invasive size, nuclear grade and estrogen receptor negativity were significantly correlated with SUV in primary breast cancer (P < 0.0001,< 0.0001, and < 0.012, respectively), and nuclear grade was significantly correlated with SUV in tumors of invasive size 2 cm or less (P < 0.0001). Tumors with high SUV (cutoff value 4.0) showed higher relapse and mortality rate compared to those with low SUV (P < 0.0001). CONCLUSIONS: High uptake of 18F-FDG would be predictive of poor prognosis in patients with primary breast cancer, and aggressive features of cancer cells in patients with early breast cancer. 18F-FDG PET/CT could be a useful tool to pre-therapeutically predict biological characteristics and baseline risk of breast cancer. PMID- 18407936 TI - Comparison of time trends in larynx cancer incidence (1973-97) in East Asia, Europe and USA, from Cancer Incidence in Five Continents, Vols. IV-VIII. PMID- 18407937 TI - Determinants of wood dust exposure in the Danish furniture industry--results from two cross-sectional studies 6 years apart. AB - OBJECTIVES: This paper investigates determinants of wood dust exposure and trends in dust level in the furniture industry of Viborg County, Denmark, using data from two cross-sectional studies 6 years apart. METHODS: During the winter 1997/1998, 54 factories were visited (hereafter study 1). In the winter 2003/2004, 27 factories were revisited, and personal dust measurements were repeated. In addition, 14 new factories were included (hereafter study 2). A total of 2303 woodworkers participated in study 1, and 2358 measurements from 1702 workers were available. From study 2, 1581 woodworkers participated and 1355 measurements from 1044 workers were available. Information on occupational variables describing potential determinants of exposures like work task, exhaust ventilation, enclosure and cleaning procedures were collected. A total of 2627 measurements and 1907 persons were included in the final mixed model in order to explore determinants of exposure and trends in dust level. RESULTS: The overall inhalable wood dust concentration (geometric means (geometric standard deviation)) has decreased from 0.95 mg/m(3) (2.05) in study 1 to 0.60 mg/m(3) (1.63) in study 2, representing a 7% annual decrease in dust concentration, which was confirmed in the mixed model. From study 1 to study 2 there has been a change towards less manual work and more efficient cleaning methods, but on the contrary also more inadequate exhaust ventilation systems. The following determinants were found to 'increase' dust concentration: sanding; use of compressed air; use of full-automatic machines; manual work; cleaning of work pieces with compressed air; kitchen producing factories and small factories (<20 employees). The following determinants of exposure were found to 'decrease' dust concentration: manual assembling/packing; sanding with adequate exhaust ventilation; adequate exhaust ventilation; vacuum cleaning of machines and special cleaning staff. CONCLUSIONS: Despite a substantial drop in the dust concentration during the last 6 years in the furniture industry in Viborg County, further improvements are possible. There should be more focus on improved exhaust ventilation, professional cleaning methods and avoiding use of compressed air. PMID- 18407938 TI - Direct observation of conformational folding coupled with disulphide rearrangement by using a water-soluble selenoxide reagent--a case of oxidative regeneration of ribonuclease A under weakly basic conditions. AB - Oxidative regeneration pathways of bovine pancreatic ribonuclease A (RNase A), which has four SS linkages, were studied at 25 degrees C and pH 8.0 by using trans-3,4-dihydroxy-1-selenolane oxide (DHS(ox)), a new selenoxide reagent with strong oxidation power. The short-term folding study using a quench-flow instrument ( approximately 1 min) revealed that early intermediates (1S, 2S, 3S and 4S) are formed stochastically and irreversibly from the reduced protein (R) and do not have any stable structures. In the long-term folding study ( approximately 300 min), on the other hand, slow generation of the key intermediates (des[65-72] and des[40-95]) through SS rearrangement from the 3S intermediate ensemble was observed, followed by slight formation of native RNase A (N). The parallel UV and CD measurements demonstrated that formation of the key intermediates is accompanied with the formation of the native-like structures. Thus, DHS(ox) allowed facile identification of the conformational folding steps coupled with SS rearrangement on the major oxidative folding pathways. PMID- 18407940 TI - Post-cardiac arrest management: more than global cooling? PMID- 18407941 TI - Volume 100: basic sciences in the British Journal of Anaesthesia. PMID- 18407943 TI - Pressure-controlled ventilation improves oxygenation during laparoscopic obesity surgery compared with volume-controlled ventilation. AB - BACKGROUND: We compared pressure and volume-controlled ventilation (PCV and VCV) in morbidly obese patients undergoing laparoscopic gastric banding surgery. METHODS: Thirty-six patients, BMI>35 kg m(-2), no major obstructive or restrictive respiratory disorder, and Pa(CO(2))<6.0 kPa, were randomized to receive either VCV or PCV during the surgery. Ventilation settings followed two distinct algorithms aiming to maintain end-tidal CO(2) (E'(CO(2))) between 4.40 and 4.66 kPa and plateau pressure (P(plateau)) as low as possible. Primary outcome variable was peroperative P(plateau). Secondary outcomes were Pa(O(2)) (Fi(O(2)) at 0.6 in each group) and Pa(CO(2)) during surgery and 2 h after extubation. Pressure, flow, and volume time curves were recorded. RESULTS: There were no significant differences in patient characteristics and co-morbidity in the two groups. Mean pH, Pa(O(2)), Sa(O(2)), and the Pa(O(2))/Fi(O(2)) ratio were higher in the PCV group, whereas Pa(CO(2)) and the E'(CO(2))-Pa(CO(2)) gradient were lower (all P<0.05). Ventilation variables, including plateau and mean airway pressures, anaesthesia-related variables, and postoperative cardiovascular variables, blood gases, and morphine requirements after the operation were similar. CONCLUSIONS: The changes in oxygenation can only be explained by an improvement in the lungs ventilation/perfusion ratio. The decelerating inspiratory flow used in PCV generates higher instantaneous flow peaks and may allow a better alveolar recruitment. PCV improves oxygenation without any side effects. PMID- 18407944 TI - Persistent train-of-four fade in myasthenia gravis patients after sevoflurane anaesthesia. PMID- 18407945 TI - Stroke volume variation obtained with FloTrac/Vigileo fails to predict fluid responsiveness in coronary artery bypass graft patients. PMID- 18407946 TI - Postoperative catheter-related pain after radical retropubic prostatectomy. PMID- 18407947 TI - Preoperative cardiopulmonary exercise testing. PMID- 18407948 TI - Does placental lactate production have a role in ephedrine-induced fetal metabolic acidosis? PMID- 18407949 TI - New use of a laryngectomy tube for management of mechanical ventilation in patients with anatomical abnormalities. PMID- 18407950 TI - Subcutaneous tetrodotoxin and inflammatory pain. PMID- 18407952 TI - Purpura in a patient with disseminated breast cancer: a rapidly progressive cancer-related thrombotic thrombocytopenic purpura. PMID- 18407953 TI - The difficult decision-making process for using or not using adjuvant chemotherapy in premenopausal endocrine-responsive breast cancer patients. PMID- 18407954 TI - Association between CYP2D6 *10 genotype and survival of breast cancer patients receiving tamoxifen treatment. AB - BACKGROUND: Human cytochrome P450 2D6 (CYP2D6) genotype may affect the efficacy of tamoxifen treatment in Caucasian women with breast cancer. The most common polymorphism of CYP2D6 in Chinese women is variant 10 (188 C to T). PATIENTS AND METHODS: Tamoxifen and 4-hydroxytamoxifen (4OHtam) were measured in the serum of 37 women with breast cancer who were receiving tamoxifen treatment. The association between CYP2D6 *10 genotype and survival was determined in a cohort of 293 women with breast cancer who received tamoxifen (n = 152) or who did not (n = 141). RESULTS: The serum 4OHtam concentrations were significantly lower in women with the CYP2D6 *10 homozygous variant T/T genotype than in those with the homozygous wild-type C/C genotype (P = 0.04). Among tamoxifen-treated women, women with the T/T genotype had a significantly worse disease-free survival (DFS) than those with the C/C or C/T genotype, and the T/T genotype remained an independent prognostic factor of DFS in multivariate analysis (hazard ratio = 4.7; 95% confidence interval = 1.1-20.0; P = 0.04). Among women who did not receive tamoxifen, there was no significant association between CYP2D6 *10 genotype and survival. CONCLUSION: In tamoxifen-treated patients, women with the CYP2D6 *10 T/T genotype have a lower 4OHtam level in the serum and a worse clinical outcome. PMID- 18407955 TI - Phase II study of sequentially administered low-dose mitomycin-C (MMC) and irinotecan (CPT-11) in women with metastatic breast cancer (MBC). AB - BACKGROUND: Preclinical studies show that mitomycin-C (MMC) followed by irinotecan (CPT-11) is synergistic. Therefore, we evaluated the toxicity and efficacy of sequentially administered low-dose MMC and CPT-11 in patients (pts) with pretreated metastatic breast cancer (MBC). Secondary objective was to evaluate the correlation between MMC-induced topoisomerase I (TOPO I) expression and NAD(P)H:quinone oxireductase 1 (NQO1) genotypes in peripheral blood mononuclear cells (PBMC) and efficacy or toxicity of the regimen. DESIGN: Thirty two pts received MMC i.v. 6 mg/m(2) day 1 and CPT-11 i.v. 125 mg/m(2) days 2 and 8 every 28 days for maximum of six cycles. TOPO I expression and NQO1 reductase genotyping in 23 of 32 (72%) pts were assayed by PCR. RESULTS: The median time to progression (TTP) was 4.7 months (95% confidence interval 4.0-5.4 months). TOPO I expression was increased 5- to 10-fold and 20- to 30-fold in PBMC at 24 and 168 h, respectively. There was no relationship between these markers and efficacy or toxicity of the regimen. CONCLUSIONS: Sequential low-dose MMC and CPT-11 was active and tolerable by pretreated MBC pts. Future trials should focus on less pretreated MBC pts and sequential tumor biopsies to test the hypothesis that increased intratumoral expression of TOPO I is related to efficacy. PMID- 18407956 TI - A multidimensional chromatography technology for in-depth phosphoproteome analysis. AB - Protein phosphorylation is a post-translational modification widely used to regulate cellular responses. Recent studies showed that global phosphorylation analysis could be used to study signaling pathways and to identify targets of protein kinases in cells. A key objective of global phosphorylation analysis is to obtain an in-depth mapping of low abundance protein phosphorylation in cells; this necessitates the use of suitable separation techniques because of the complexity of the phosphoproteome. Here we developed a multidimensional chromatography technology, combining IMAC, hydrophilic interaction chromatography, and reverse phase LC, for phosphopeptide purification and fractionation. Its application to the yeast Saccharomyces cerevisiae after DNA damage led to the identification of 8764 unique phosphopeptides from 2278 phosphoproteins using tandem MS. Analysis of two low abundance proteins, Rad9 and Mrc1, revealed that approximately 50% of their phosphorylation was identified via this global phosphorylation analysis. Thus, this technology is suited for in depth phosphoproteome studies. PMID- 18407957 TI - Comparative proteomics analysis reveals an intimate protein network provoked by hydrogen peroxide stress in rice seedling leaves. AB - Hydrogen peroxide (H2O2) plays a dual role in plants as the toxic by-product of normal cell metabolism and as a regulatory molecule in stress perception and signal transduction. However, a clear inventory as to how this dual function is regulated in plants is far from complete. In particular, how plants maintain survival under oxidative stress via adjustments of the intercellular metabolic network and antioxidative system is largely unknown. To investigate the responses of rice seedlings to H2O2 stress, changes in protein expression were analyzed using a comparative proteomics approach. Treatments with different concentrations of H2O2 for 6 h on 12-day-old rice seedlings resulted in several stressful phenotypes such as rolling leaves, decreased photosynthetic and photorespiratory rates, and elevated H2O2 accumulation. Analysis of approximately 2000 protein spots on each two-dimensional electrophoresis gel revealed 144 differentially expressed proteins. Of them, 65 protein spots were up-regulated, and 79 were down regulated under at least one of the H2O2 treatment concentrations. Furthermore 129 differentially expressed protein spots were identified by mass spectrometry to match 89 diverse protein species. These identified proteins are involved in different cellular responses and metabolic processes with obvious functional tendencies toward cell defense, redox homeostasis, signal transduction, protein synthesis and degradation, photosynthesis and photorespiration, and carbohydrate/energy metabolism, indicating a good correlation between oxidative stress-responsive proteins and leaf physiological changes. The abundance changes of these proteins, together with their putative functions and participation in physiological reactions, produce an oxidative stress-responsive network at the protein level in H2O2-treated rice seedling leaves. Such a protein network allows us to further understand the possible management strategy of cellular activities occurring in the H2O2-treated rice seedling leaves and provides new insights into oxidative stress responses in plants. PMID- 18407959 TI - Immunolocalization of water channel aquaporins in the vomeronasal organ of the rat: expression of AQP4 in neuronal sensory cells. AB - The vomeronasal organ comprises a pair of narrow tubes in the mammalian nasal septum, serving as a chemosensory system for pheromones. We examined the expression and localization of water channel aquaporins (AQPs) in the rat vomeronasal organ. AQP1 was localized in blood vessels, being particularly abundant in cavernous tissues of the nonsensory mucosa. AQP5 was found in the apical membrane of the gland acinar cells in the vomeronasal organ. AQP3 was detected in the basal cells of the nonsensory epithelium, whereas it was absent in the sensory epithelium. AQP4 was found in both the sensory and the nonsensory epithelia. Interestingly, AQP4 was highly concentrated in the sensory cells of the sensory epithelium. Immunoelectron microscopic examination clearly showed that AQP4 was localized at the plasma membrane in the cell body and lateral membrane of the dendrite, except for the microvillous apical membrane. Nerve fiber bundles emanating from neuronal sensory cells were positive for AQP4, whereby the plasma membrane of each axon was positive for AQP4. These observations clearly show that neuronal sensory cells in the vomeronasal organ are unique in that they express abundant AQP4 at their plasma membrane. This is in marked contrast to the olfactory and central nervous systems, where AQPs are not detectable in neurons, and instead, AQP4 is abundant in the supporting cells and astrocytes surrounding them. The present findings suggest a unique water handling feature in neuronal sensory cells in the vomeronasal organ. PMID- 18407958 TI - Monitoring protein-protein interactions between the mammalian integral membrane transporters and PDZ-interacting partners using a modified split-ubiquitin membrane yeast two-hybrid system. AB - PDZ-binding motifs are found in the C-terminal tails of numerous integral membrane proteins where they mediate specific protein-protein interactions by binding to PDZ-containing proteins. Conventional yeast two-hybrid screens have been used to probe protein-protein interactions of these soluble C termini. However, to date no in vivo technology has been available to study interactions between the full-length integral membrane proteins and their cognate PDZ interacting partners. We previously developed a split-ubiquitin membrane yeast two-hybrid (MYTH) system to test interactions between such integral membrane proteins by using a transcriptional output based on cleavage of a transcription factor from the C terminus of membrane-inserted baits. Here we modified MYTH to permit detection of C-terminal PDZ domain interactions by redirecting the transcription factor moiety from the C to the N terminus of a given integral membrane protein thus liberating their native C termini. We successfully applied this "MYTH 2.0" system to five different mammalian full-length renal transporters and identified novel PDZ domain-containing partners of the phosphate (NaPi-IIa) and sulfate (NaS1) transporters that would have otherwise not been detectable. Furthermore this assay was applied to locate the PDZ-binding domain on the NaS1 protein. We showed that the PDZ-binding domain for PDZK1 on NaS1 is upstream of its C terminus, whereas the two interacting proteins, NHERF-1 and NHERF-2, bind at a location closer to the N terminus of NaS1. Moreover NHERF-1 and NHERF-2 increased functional sulfate uptake in Xenopus oocytes when co-expressed with NaS1. Finally we used MYTH 2.0 to demonstrate that the NaPi-IIa transporter homodimerizes via protein-protein interactions within the lipid bilayer. In summary, our study establishes the MYTH 2.0 system as a novel tool for interactive proteomics studies of membrane protein complexes. PMID- 18407960 TI - Should surgical pleurectomy for spontaneous pneumothorax be always thoracoscopic? AB - Fifty-seven patients were studied over a period of three years to analyse the efficacy of surgical pleurectomy for spontaneous pneumothorax. Thirty-one and 26 patients underwent open and video-assisted thoracoscopic surgery (VATS) pleurectomy, respectively. VATS was the main modality used for primary spontaneous pneumothorax (PSP) (21 vs. 8). However, secondary spontaneous pneumothorax (SSP) was mainly managed with open pleurectomy (23 vs. 5). The median operating time was significantly longer in open group (72.4 vs. 55 min; P=0.005). The amount of analgesia required in the first five days was significantly more in open group (108 mg vs. 46.9 mg; P=0.02). Chest drainage was significantly more in open group (1027.1 ml vs. 652.8 ml; P=0.04). However, chest drain duration and hospital stay had no significant difference. VATS emerged as a cost-effective modality (1770 pounds vs. 3226 pounds). The ability to return to work was significantly earlier in VATS group in PSP patients (6 weeks vs. 10 weeks; P=0.007). There were 3 (5.27%) recurrences in VATS group for patients with SSP. This experience suggests that VATS pleurectomy is an appropriate modality for PSP. However, open pleurectomy is a viable alternative to treat SSP. PMID- 18407961 TI - Is a port-access mitral valve repair superior to the sternotomy approach in accelerating postoperative recovery? AB - A best evidence topic in cardiac surgery was written according to a structured protocol. The question addressed was whether port-access mitral valve repair reduces the recovery period of patients compared to the conventional sternotomy approach. Using the reported search, 778 papers were identified. Thirteen papers represented the best evidence on the subject and the author, journal, date and country of publication, patient group studied, study type, relevant outcomes, results and study comments and weaknesses were tabulated. The 13 papers demonstrated that patients who undergo minimally invasive mitral valve repair have a shorter ICU and total hospital stay than those who undergo the sternotomy approach. Results vary but mean hospital stays range from 5.6 to 13 days in port access groups compared to 6.25-15 days in sternotomy groups. Other advantages over the sternotomy approach were reduced postoperative bleeding and pain, shorter time to extubation and a quicker return to daily activities. However, it is consistently reported that operative time is longer, with the increase in bypass time being around 30 min. We conclude that in several cohort studies minimally invasive mitral valve repair is reported to result in a shorter ICU and hospital stay, reduced postoperative bleeding and pain and a shorter time to resuming normal activities. This is at the expense of longer bypass and operative times. PMID- 18407962 TI - Rare venous connection causing severe hypoxia after Fontan operation. AB - We describe a rare case of cyanosis following the Fontan operation with right-to left shunting at the venous level, that is, an azygos venous to pulmonary venous connection. Few cases with partial anomalous pulmonary venous connection to azygos vein have been reported; however, there have been no reports describing the connection from azygos vein to pulmonary vein, which results in desaturation after Fontan operation. Multidetector-row computed tomography (MDCT) was a useful tool to depict this vascular malformation. PMID- 18407963 TI - Elastofibroma of the thoracic wall. AB - Soft tissue tumors of the chest wall are rare. Between 1998 and 2007 we treated eight cases of elastofibroma of the thoracic wall, an infrequent primary tumor of the chest. Seven females and one male between 44 and 62 years presented with dorsal subscapular tumors of months and even years of evolution. One case was a relapse from previous interventions and in three cases the tumor was bilateral. A surgical excision was performed in all cases, confirming the source of the tumor. There were no postoperative complications or relapses. We concluded that elastofibroma is a tumor that appears most frequently in middle aged women, and that diagnosis can be established through the use of imaging and fine needle aspiration biopsy. Given its benign character and slow growth, in cases where it is asymptomatic, its evolution can be controlled without surgical intervention. PMID- 18407964 TI - Evidence for sugar signalling in the regulation of asparagine synthetase gene expressed in Phaseolus vulgaris roots and nodules. AB - A cDNA clone, designated as PvNAS2, encoding asparagine amidotransferase (asparagine synthetase) was isolated from nodule tissue of common bean (Phaseolus vulgaris cv. Negro Jamapa). Southern blot analysis indicated that asparagine synthetase in bean is encoded by a small gene family. Northern analysis of RNAs from various plant organs demonstrated that PvNAS2 is highly expressed in roots, followed by nodules in which it is mainly induced during the early days of nitrogen fixation. Investigations with the PvNAS2 promoter gusA fusion revealed that the expression of PvNAS2 in roots is confined to vascular bundles and meristematic tissues, while in root nodules its expression is solely localized to vascular traces and outer cortical cells encompassing the central nitrogen-fixing zone, but never detected in either infected or non-infected cells located in the central region of the nodule. PvNAS2 is down-regulated when carbon availability is reduced in nodules, and the addition of sugars to the plants, mainly glucose, boosted its induction, leading to the increased asparagine production. In contrast to PvNAS2 expression and the concomitant asparagine synthesis, glucose supplement resulted in the reduction of ureide content in nodules. Studies with glucose analogues as well as hexokinase inhibitors suggested a role for hexokinase in the sugar-sensing mechanism that regulates PvNAS2 expression in roots. In light of the above results, it is proposed that, in bean, low carbon availability in nodules prompts the down-regulation of the asparagine synthetase enzyme and concomitantly asparagine production. Thereby a favourable environment is created for the efficient transfer of the amido group of glutamine for the synthesis of purines, and then ureide generation. PMID- 18407965 TI - In vivo role of Escherichia coli single-strand exonucleases in SOS induction by gamma radiation. AB - Ionizing radiation causes different types of genetic damage, ranging from base modifications to single- and double-stranded DNA breaks, which may be deleterious or even lethal to the cell. There are different repair or tolerance mechanisms to counteract the damage. Among them is the Escherichia coli SOS system: a set of genes that becomes activated upon DNA damage to confer better opportunities for cell survival. However, since this response is triggered by single-stranded DNA regions, most lesions have to be processed or modified prior to SOS activation. Several genes such as recO, recB and recJ that seem to be required to induce the response have already been reported. The results of this work indicate that the four known E.coli single-strand exonucleases take part in processing gamma radiation damage, though RecJ and ExoI proved to be more important than ExoVII or ExoX. In addition, ExoV as well as glycosylases such as Nth and, to a lesser extent, Fpg are also required. A model intended to explain the role of all these genes in damage processing is presented. PMID- 18407966 TI - Acrylamide-induced molecular mutation spectra at HPRT locus in human promyelocytic leukaemia HL-60 and NB4 cell lines. AB - Acrylamide (AA) is a compound widely used in many industries around the world. The recent finding that it is formed naturally in foods by heating raises human health concerns. AA is a proven carcinogen in animals and a probable carcinogen in humans, while its mutagenicity detected using in vitro mammalian gene mutation assays is still inconsistent in different cell systems. In the present study, we investigated the mutagenicity of AA in human promyelocytic leukaemia cells, HL-60 and NB4 cells, by examining the mutations at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene locus. In a 6-h treatment without the exogenous activation, AA exerted a weak mutagenic effect at the highest concentration used in the study (700 mg/l) in HL-60 cells (P < 0.01) as well as in NB4 cells (P < 0.05). Molecular analysis of AA-induced mutants revealed a different mutation spectrum, when compared to that of spontaneous mutants. The most frequent spontaneous mutations were point mutations, whereas AA-induced mutations were mainly single exon deletions besides point mutations, and an increase in the proportion of partial deletion was associated with the increase of AA treatment. There was no obvious difference in the mutation spectra observed between the HL-60 and NB4 cell lines. These results showed that AA has a weak mutagenic effect at HPRT gene locus in human promyelocytic leukaemia HL-60 and NB4 cell lines and those molecular mutation spectra (single exon deletions and point mutations) may be related to some specific and precise mechanism. PMID- 18407967 TI - Activation-induced NKT cell hyporesponsiveness protects from alpha galactosylceramide hepatitis and is independent of active transregulatory factors. AB - NK T (NKT) cells, unique lymphocytes expressing features of NK and T lymphocytes, can specifically be activated with the glycolipid antigen alpha galactosylceramide (alpha-GalCer). In humans and mice, this activation provokes pronounced cytokine responses. In C57BL/6 mice, alpha-GalCer injection additionally induces NKT-mediated liver injury, representing a model for immune mediated hepatitis in humans. However, a single alpha-GalCer pretreatment of mice prevented NKT-mediated liver injury, cytokine responses (systemically and locally in the liver), and up-regulation of hepatocellular Fas upon alpha-GalCer rechallenge. As alpha-GalCer is used as a NKT cell-activating agent in clinical trials, an investigation of tolerance induction appears crucial. We demonstrate that alpha-GalCer tolerance does not depend on Kupffer cells, IL-10, Caspase-3 mediated apoptosis, or CD4+CD25+ T regulatory cells (Tregs), which are crucial in other models of immunological tolerance. Amending relevant, earlier approaches of others, we cocultivated highly purified, nontolerized and tolerized liver NKT cells ex vivo and could convincingly exclude the relevance of transdominant NKT Tregs. These results strongly suggest alpha-GalCer-induced tolerance to be exclusively caused by NKT cell intrinsic hyporesponsiveness. Tolerized mice showed specific diminishment of the intrahepatic CD4+ NKT cell subpopulation, with the CD4(-) population largely unaffected, and revealed down-modulation of alpha-GalCer-specific TCR and the NKT costimulator glucocorticoid-induced TNFR related protein on liver NKT cells, whereas inhibitory Ly49I was increased. In conclusion, alpha-GalCer tolerance could serve as a model for the frequently observed NKT cell hyporesponsiveness in tumor patients and might help to develop strategies for their reactivation. Conversely, approaches to render NKT cells hyporesponsive may constitute new therapeutic strategies for diseases, where aberrant NKT cell activation is causally involved. PMID- 18407968 TI - Cytokines and fibrocyte differentiation -- altering the balance: an interview with Dr. Darrell Pilling. Interview by Helene F. Rosenberg. PMID- 18407969 TI - Direct His bundle pacing preserves coronary perfusion compared with right ventricular apical pacing: a prospective, cross-over mid-term study. AB - AIMS: The His bundle is regarded as the most physiological site for ventricular pacing, in that it avoids the adverse effects of right ventricular apical pacing (RVAP). However, very few studies have compared the effects of direct His bundle pacing (DHBP) and RVAP. The aim of our study was the intra-patient comparison of myocardial perfusion corresponding to these two different pacing techniques, as perfusion expresses local workload and is related to long-term outcome. METHODS AND RESULTS: Twelve consecutive patients with standard pacemaker indication (9 male, 74 +/- 9 years) entered the study. Pacing leads were implanted in the right ventricular apex and directly in the His bundle, and were connected to different ports of the pacemaker. All patients first underwent 3 months of DHBP, followed by 3 months of RVAP. At the end of each 3-month period, myocardial perfusion was measured at rest using scintigraphy with Tc99m-SestaMIBI. The average values of perfusion were evaluated on a 20-segment basis. All patients also underwent clinical evaluation, echocardiography, and tissue Doppler imaging (TDI), to measure dyssynchrony, and a blood sample was taken for brain natriuretic peptide (BNP) assay. The perfusion score during DHBP pacing was significantly better than during RVAP (0.44 +/- 0.5 vs. 0.71 +/- 0.53, respectively; P = 0.011). None of the patients showed lower perfusion during DHBP than during RVAP. We found no significant difference in NYHA class, ventricular volumes, ejection fraction, or plasmatic BNP between DHBP and RVAP. However, mitral regurgitation (0.26 +/- 0.21 vs. 0.37 +/- 0.25; P < 0.001) and dyssynchrony (13.75 +/- 4.28 vs. 22.02 +/- 8.44; P = 0.008) were significantly less during DHBP than during RVAP. CONCLUSION: Direct His bundle pacing is superior to RVAP in preserving the physiologic distribution of myocardial blood flow and reducing mitral regurgitation and left ventricular dyssynchrony. PMID- 18407970 TI - Rapid comparative genomic analysis for clinical microbiology: the Francisella tularensis paradigm. AB - It is critical to avoid delays in detecting strain manipulations, such as the addition/deletion of a gene or modification of genes for increased virulence or antibiotic resistance, using genome analysis during an epidemic outbreak or a bioterrorist attack. Our objective was to evaluate the efficiency of genome analysis in such an emergency context by using contigs produced by pyrosequencing without time-consuming finishing processes and comparing them to available genomes for the same species. For this purpose, we analyzed a clinical isolate of Francisella tularensis subspecies holarctica (strain URFT1), a potential biological weapon, and compared the data obtained with available genomic sequences of other strains. The technique provided 1,800,530 bp of assembled sequences, resulting in 480 contigs. We found by comparative analysis with other strains that all the gaps but one in the genome sequence were caused by repeats. No new genes were found, but a deletion was detected that included three putative genes and part of a fourth gene. The set of 35 candidate LVS virulence attenuation genes was identified, as well as a DNA gyrase mutation associated with quinolone resistance. Selection for variable sequences in URFT1 allowed the design of a strain-specific, highly effective typing system that was applied to 74 strains and six clinical specimens. The analysis presented herein may be completed within approximately 6 wk, a duration compatible with that required by an urgent context. In the bioterrorism context, it allows the rapid detection of strain manipulation, including intentionally added virulence genes and genes that support antibiotic resistance. PMID- 18407971 TI - Detection properties of a measuring system for a continuous soil radon concentrations monitoring. AB - The continual soil-gas radon concentration measurements are absolutely crucial for a reliable assessment of radon entry characteristics into the indoor building environment. For this purpose, a new detection system (a continuous monitor RM-3) was developed and tested. The detection principle of the monitoring device is based on an airflow ionisation chamber operating in a current mode. A comprehensive series of testing and calibration experiments have been carried out in a laboratory environment. An output signal of the device caused by the radon concentration in a sensitive detection volume significantly depends on a detector ventilation rate, the gas flow rate through the ionisation chamber. A set of calibration experiments was accomplished with the artificial radon source application and close circuit experimental arrangements. The system detection properties including applied experimental conditions and key results of pilot in situ measurements are reported in detail. PMID- 18407972 TI - Assessment of inhomogeneous ELF magnetic field exposures. AB - In daily life as well as at workplaces, exposures to inhomogeneous magnetic fields become very frequent. This makes easily applicable compliance assessment methods increasingly important. Reference levels have been defined linking basic restrictions to levels of homogeneous fields at worst-case exposure conditions. If reference levels are met, compliance with basic restrictions can be assumed. If not, further investigations could still prove compliance. Because of the lower induction efficiency, inhomogeneous magnetic fields such as from electric appliances could be allowed exceeding reference levels. To easily assess inhomogeneous magnetic fields, a quick and flexible multi-step assessment procedure is proposed. On the basis of simulations with numerical, anatomical human models reference factors were calculated elevating reference levels to link hot-spot values measured at source surfaces to basic limits and allowing accounting for different source distance, size, orientation and position. Compliance rules are proposed minimising assessment efforts. PMID- 18407973 TI - Human perception of radon risk and radon mitigation: some remarks. AB - The Radon program in the Czech Republic has a relatively long and rich history. Procedures, which enable to evaluate the risk of radon penetration from the ground, to protect new buildings, to find existing buildings with elevated indoor radon levels and to realise remedial measures in such buildings, have been developed, published and tested. In some cases, the whole system may fail due to psychological or sociological reasons. Three types of problems (conflicts) will be presented: human behaviour affecting measurement results, conflict between individual and 'all-society' points of view, interpretation of radon risk itself. PMID- 18407974 TI - Evaluation of occupational exposures to natural radiation in the Czech Republic. AB - The Czech approach to limit the occupational exposures to natural radiation is based on the rules given by the Atomic Act and by the Decree of the State Office for Nuclear Safety (SONS) on radiation protection. Workplaces with potential risk of increased exposure to natural sources are specified explicitly. A new method to perform the above-mentioned measurements and to determine the effective doses of workers was proposed in 2005 and accepted by the SONS in 2007. The first experience illustrates its applicability. PMID- 18407975 TI - Radon diagnosis procedures focused on dwellings with ineffective measures against radon. AB - An efficiency analysis of remedial measures launching in the Czech Republic before 2004 revealed that in 52% of houses the remediation was not successful. An example of the house with an ineffective mitigation is described in order to present radon diagnosis procedures focused on the identification of critical factors responsible for the remedial mitigations insufficiency. A description of additional measures application is presented and results of the final efficiency evaluation are discussed. PMID- 18407976 TI - Can the HML's sliced BOMAB phantom be used in any whole body counter with a reduced number of sources? AB - The sliced Bottle Manikin Absorber (BOMAB) phantom was originally proposed as an alternative to a commercially available phantom, but it suffers from the disadvantage of containing over 160 sources that need to be manufactured; however, it was found that the number of slices could be reduced substantially and that two slices in the sliced phantom gave the same performance characteristics over a wide energy range as a conventional BOMAB phantom for a particular counting system. This work explores the adaptability of this phantom to another counting geometry. The response of the Human Monitoring Laboratory's whole-body counter measuring this phantom with a decreasing number of planar sources has been modelled using MCNP5 over a wide energy range (122-2754 keV). It was found that the best agreement was obtained when the phantom contained 10 sources, 1 in the mid point of each section. As this is a different result from a previous finding, any other counting geometry will have to be assessed to determine the optimum loading if the sliced phantom is to be used. Also, it is clear that this type of phantom cannot be used for an intercomparison that will encounter different counting geometries, unless it contains a full loading of sources. PMID- 18407977 TI - Responses to delayed estrus after weaning in sows using oral progestagen treatment. AB - Oral progestagen treatment extends the weaning-to-estrus interval (WEI) in weaned sows. Particularly in lower parity sows, this allows recovery from lactational catabolism and improves sow productivity. However, the optimal duration of progestagen treatment in contemporary dam-line sows is unclear. Therefore, sows (n = 749) weaned over consecutive 3-wk periods in June and July and classified as parity 2 and 3 (P2-3); 4, 5, and 6 (P4-6); or parity 7 or higher (P7+) were organized into 2 breeding groups using 1 of 3 strategies: 1) oral progestagen for 2 d before and 12 d after weaning (M14; n = 249); 2) oral progestagen for 2 d before and 5 d after weaning (M7; n = 250); or 3) no progestagen treatment (M0; n = 250). Progestagen (altrenogest) was administered directly into the sow's mouth at a dosage of 6.8 mL (15 mg of altrenogest) daily. Sows were bred using artificial insemination at first detection of estrus after weaning (M0) or altrenogest withdrawal, and every 24 h thereafter, until they no longer exhibited the standing reflex. The WEI for M0 sows was 5.1 +/- 0.1 d. Estrus was recorded sooner (P < 0.001) after withdrawing treatment in M14 than in M7 sows (6.9 +/- 0.1 vs. 7.4 +/- 0.1 d, respectively). More (P < 0.001) M14 sows (88.6 +/- 2.5%) were bred within 10 d of altrenogest withdrawal than M7 (72.8 +/- 2.8%) sows, or within 10 d of weaning in M0 sows (78.8 +/- 2.6%). Reproductive tracts were recovered after slaughter at d 30 or 50 of gestation. For P2-3 sows, ovulation rate (least squares mean +/- 95% confidence interval) in M7 (23.1 +/- 1.0) was greater (P < 0.001) than in M14 (20.7 +/- 1.0) or M0 (19.7 +/- 1.0) sows; no differences were detected in P4-6 and P7+ sows. At d 30, M7 and M14 sows had more (P < 0.01) embryos (16.4 +/- 0.6 and 15.8 +/- 0.4, respectively) than M0 (13.9 +/ 0.5) sows. At d 50 of gestation, number of fetuses in M14 sows (13.6 +/- 0.4) was greater (P < 0.001) than in M0 (11.8 +/- 0.4) and M7 (12.2 +/- 0.3) sows. Use of oral progestagen to delay the return to postweaning estrus for greater than 18 d appears to have potential for improving weaned sow productivity. Given the incidence of high ovulation rates and associated evidence of intrauterine crowding of embryos around d 30 of gestation, the changing dynamics of prenatal loss resulting from longer periods of progestagen treatment may represent an additional production advantage. PMID- 18407978 TI - Association of the prion protein gene with individual tissue weights in Scottish Blackface sheep. AB - This study investigated associations of prion protein (PrP) genotype with body composition and weight traits of Scottish Blackface ewes. Body composition was predicted using computer tomography (CT) scans to estimate muscle, carcass fat, internal fat, and bone weights. The traits were measured at 4 key seasonal production points (pre-mating, pregnancy, midlactation, and weaning) over 4 production cycles (2 to 5 yr old). There were 2,413 records for each of the CT traits measured on 335 ewes, and 26,649 records for each of the body condition score and BW traits for 2,356 ewes. From 1999 to 2004, animals were genotyped to determine polymorphisms at codons 136, 154, and 171, which are associated with scrapie susceptibility. Four alleles were found in the population (ARR, AHQ, ARQ, and VRQ). The data were analyzed using a linear mixed random regression model assuming that the direct additive genetic effect was a 2nd order Legendre polynomial function of time. The PrP genotype was included in the model as a fixed effect along with other fixed factors with significant effects (P < 0.05). Five separate analyses were carried out for each trait, depending on the method of classifying the PrP genotype. In the first analysis, animals were categorized according to the genotype. Only the 5 most common genotypes (ARR/ARR, ARR/AHQ, ARR/ARQ, AHQ/ARQ, and ARQ/ARQ) were included. In the last 4 analyses, animals were categorized according to the number of each PrP allele carried. For CT traits and body condition score, results showed that the PrP genotype has no association with the overall mean of the traits (averaged over age). For BW, ewes without the ARQ allele were at least 0.5 kg heavier than ARQ homozygous and heterozygous ewes. On the other hand, there was a significant interaction between PrP genotype and age of the ewe (i.e., the effect of PrP genotype was not the same at different ages for 5 out of the 6 traits studied). In general, ARQ carrying ewes mobilized more fat reserves at times of nutrient deficiency, such as during lactation, and gained it back more quickly by the mating season (when nutrients became abundant) than non-ARQ carriers. Therefore, selecting against this allele would have consequences on BW and seasonal mobilization of body reserves. The number of VRQ alleles (the most scrapie susceptible allele) carried was not significantly associated with any of the traits. PMID- 18407979 TI - Effects of dietary organic and inorganic trace mineral levels on sow reproductive performances and daily mineral intakes over six parities. AB - Dietary trace mineral sources and levels were fed to developing gilts to evaluate their performance responses during the growth phase, but treatments were continued into the reproductive phase in which subsequent reproductive responses were evaluated. In Exp. 1, three groups of gilts (n = 216) were used in a 2 x 2 factorial in a randomized complete block design (6 replicates) with treatment diets initially fed at 30 kg of BW. The first factor was trace mineral source (organic or inorganic), whereas the second factor evaluated dietary levels. The NRC requirement was the first level evaluated, whereas the second level was formulated to average industry standards (IND). Organic trace minerals were mineral proteinates, whereas the inorganic minerals were provided in salt form. The results of Exp. 1 indicated that trace mineral source or level did not affect gilt growth or feed performance responses to 110 kg of BW. Experiment 2 continued with the same females but was a 2 x 3 factorial in a split-plot design using 3 groups of females over a 6-parity period and had a total of 375 farrowings. Factors in Exp. 2 were the same as in Exp. 1, except that 2 additional pens of gilts during their development had been fed the IND level trace mineral levels of both trace mineral sources. At breeding, the gilts from these 2 additional pens were continued on the same trace mineral source and level but fed greater dietary Ca and P levels (IND + Ca:P). Litters were standardized by 3 d postpartum within each farrowing. Sows fed organic trace minerals farrowed more (P < 0.05) total (12.2 vs. 11.3) and live pigs (11.3 vs. 10.6) compared with sows fed inorganic trace minerals. Sows fed the IND + Ca:P level tended to have fewer (P < 0.10) total pigs born for both trace mineral sources. Litter birth weights were heavier (P < 0.05) when sows were fed organic trace minerals, but individual piglet weights were similar. Nursing pig ADG tended to be greater (P < 0.10) when sows were fed organic trace minerals. Other sow reproductive traits (BW, feed intake, and rebreeding interval) were not affected by trace mineral source or level. Daily mineral intake increased by parity but declined when trace mineral intakes were expressed on an amount per kilogram of BW and declined during later lactations. These results suggest that feeding sows organic trace minerals may improve sow reproductive performance, but there were minimal effects on other reproductive measurements. PMID- 18407980 TI - Genomic selection using different marker types and densities. AB - With the availability of high-density marker maps and cost-effective genotyping, genomic selection methods may provide faster genetic gain than can be achieved by current selection methods based on phenotypes and the pedigree. Here we investigate some of the factors driving the accuracy of genomic selection, namely marker density and marker type (i.e., microsatellite and SNP markers), and the use of marker haplotypes versus marker genotypes alone. Different densities were tested with marker densities equivalent to 2, 1, 0.5, and 0.25N(e) markers/morgan using microsatellites and 8, 4, 2, and 1N(e) markers/morgan using SNP, where 1N(e) markers/morgan means 100 markers per morgan, if effective size (N(e)) is 100. Marker characteristics and linkage disequilibria were obtained by simulating a population over 1,000 generations to achieve a mutation drift balance. The marker designs were evaluated for their accuracy of predicting breeding values from either estimating marker effects or estimating effects of haplotypes based upon combining 2 markers. Using microsatellites as direct marker effects, the accuracy of selection increased from 0.63 to 0.83 as the density increased from 0.25N(e)/morgan to 2N(e)/morgan. Using SNP markers as direct marker effects, the accuracy of selection increased from 0.69 to 0.86 as the density increased from 1N(e)/morgan to 8N(e)/morgan. The SNP markers required a 2 to 3 times greater density compared with using microsatellites to achieve a similar accuracy. The biases that genomic selection EBV often show are due to the prediction of marker effects instead of QTL effects, and hence, genomic selection EBV may need rescaling for practical use. Using haplotypes resulted in similar or reduced accuracies compared with using direct marker effects. In practical situations, this means that it is advantageous to use direct marker effects, because this avoids the estimation of marker phases with the associated errors. In general, the results showed that the accuracy remained responsive with small bias to increasing marker density at least up to 8N(e) SNP/morgan, where the effective population size was 100 and with the genomic model assumed. For a 30-morgan genome and N(e) = 100, this implies that about approximately 24,000 SNP are needed. PMID- 18407981 TI - Effects of direct-fed microbial supplementation on digestibility and fermentation end-products in horses fed low- and high-starch concentrates. AB - A study was conducted to determine whether direct-fed microbials (DFM) could be used to increase digestibility and minimize the risk of acidosis associated with feeding an increase in the amount of starch fed to horses. Fifteen mature Thoroughbred geldings were randomly assigned to 1 of 3 treatments in a 3 x 3 Latin square design balanced for carryover effects. Within each 26-d period, horses were offered grass hay + low-starch concentrate (LS; 1.2 g of starch x kg of BW(-1) x meal(-1)) from d 1 to 13 and then were abruptly changed to hay + high starch concentrate (HS; 2.4 g of starch.kg of BW(-1)x meal(-1)) on d 14 continuing through d 26. The DFM treatments were offered in concentrate pellets at a target dosage of 10(8) cfu/(50 kg of BW x d) as follows: no DFM (CON; control), Lactobacillus acidophilus (LAC1; single-species DFM), or a mixture of L. acidophilus, Lactobacillus casei, Bifidobacterium bifidum, and Enterococcus faecium (LAC4; multiple-species DFM). Total feces were collected over 72 h from d 11 to 13 (LS; low dietary starch collection), from d 15 to 17 (AC; abrupt change in dietary starch collection), and at the end of each experimental period, from d 24 to 26 (HS; high dietary starch collection). Data collected consisted of total DM intake and fecal output, fecal pH, fecal acetate and propionate concentrations, and viable numbers of DFM in the feed. With the exception of Fe digestibility, there were no starch x DFM interactions. There was an effect of starch level (P 0.10). Horses supplemented with LAC4 had increased ether extract (P < 0.05) and a tendency for decreased Na (P < 0.10) digestibilities compared with CON horses. All DFM-supplemented horses had increased Cu (P < 0.05) and Fe and numerically increased Zn digestibilities compared with CON horses. Fecal pH decreased (P < 0.05), and fecal propionate concentration increased (P < 0.05) as dietary starch content changed from LS to HS. There was a tendency for elevated fecal pH (P < 0.10) in LAC1 horses compared with CON horses. These results confirm that increasing starch in the equine diet can enhance nutrient digestibility of the diet. Supplementing equine diets with either a single or mixed strain direct-fed lactic acid bacteria had limited effects on nutrient digestibility or on reducing the risk of acidosis associated with feeding high-starch concentrates to horses. The potential response of DFM supplementation should be evaluated when a more acute acidotic state is induced in horses than in the current study. PMID- 18407982 TI - Technical note: prediction of breeding values using marker-derived relationship matrices. AB - In livestock populations, estimation of breeding values for selection requires a matrix describing the additive relationship between individuals in the population. This matrix can be derived from pedigree information. In some livestock populations, pedigree information may be unavailable, incomplete, or in error. Here we use simulated data to demonstrate that marker-derived relationship matrices can be used to predict breeding values and estimate additive variance components, provided the markers are sufficiently dense. The approach is demonstrated for an Angus data set with 9,323 SNP markers genotyped. PMID- 18407983 TI - Effects of 2-hydroxy-4-(methylthio)-butanoic acid on performance and carcass characteristics of finishing beef cattle and on fermentation in continuous culture. AB - Three experiments were conducted to evaluate the effects of feeding 2-hydroxy-4- (methylthio)-butanoic acid (HMTBA) on performance and carcass characteristics of feedlot cattle and on microbial fermentation in a continuous-culture system. In Exp. 1, 160 crossbred steers (initial BW = 385 +/- 10.3 kg) were assigned to 4 treatments consisting of control (0% HMTBA) or 3 diets containing HMTBA (0.069, 0.137, and 0.204%; DM basis) in a randomized complete block design. As the percent of HMTBA increased in the diet, final BW (P = 0.069), final BW adjusted to a constant dressing percent (P = 0.063), and overall ADG (P = 0.099) tended to decrease linearly. Overall DMI decreased linearly (P < or = 0.006) with increasing HMTBA dose. No differences (P > or = 0.10) were noted for carcass characteristics, except for a tendency (P = 0.078) for a linear increase in the percentage of cattle grading USDA Choice with increasing HMTBA dose. In Exp. 2, 80 crossbred steers (initial BW = 450 +/- 17 kg) in a randomized complete block design were assigned to a control (0% HMTBA) diet or to a diet in which the concentrations of HMTBA were gradually increased from 0.036 to 0.212% of DM over a 50-d period. The HMTBA-containing diet tended to decrease DMI (P = 0.132), but G:F (P = 0.319) for the overall feeding period, carcass measurements, and USDA quality grade (P > or = 0.149) did not differ between treatments. In Exp. 3, continuous culture fermenters (n = 5/treatment) were used to determine the effects of HMTBA (control vs. 0.24% HMTBA) on microbial fermentation. No differences (P > or = 0.31) were detected between treatments in ruminal OM digestibility, microbial N synthesis, pH, ammonia, molar proportions of VFA, or effluent concentration of selected long-chain fatty acids. These results suggest that HMTBA decreased DMI by feedlot steers fed a steam-flaked corn-based diet in a dose-dependent manner; however, gradually increasing the dose over time seemed to moderate effects on DMI. No major changes in microbial fermentation in continuous culture were observed with HMTBA at 0.24% of dietary DM, suggesting effects of HMTBA on DMI were not likely associated with changes in ruminal digestion or fermentation. PMID- 18407984 TI - Changes in food intake and circulating leptin due to gastrointestinal parasitism in lambs of two breeds. AB - A reduction in food intake is a prominent feature of many infectious diseases. However, the underlying mechanisms of parasite-induced anorexia in sheep are poorly understood. Here, we tested the hypotheses (a) that the degree of parasite induced anorexia in lambs is influenced by their growth potential and (b) that nematode infection results in elevated plasma leptin concentration in lambs. The hypotheses were tested with Suffolk x Greyface (S) and Scottish Black-face (B) lambs that are known to differ in their growth potential (S lambs are of greater growth potential than B lambs). During a primary parasite infection, 24 out of 48 lambs per breed were trickle-infected with 7,000 infective Teladorsagia circumcincta larvae per day, 3 d/wk, for a period of 12 wk (experiment I). The lambs were then dewormed, and after a 2-wk interval, half of the 24 lambs per breed that were previously infected were reinfected for another 12 wk with the same parasite and dose as used in the primary infection (experiment II). In both experiments, infected lambs were fed grass pellets for ad libitum intake, whereas noninfected lambs were fed grass pellets for either ad libitum or restricted intakes. The S lambs were more susceptible than B lambs to nematode infection, as judged from the differences in fecal egg counts (P = 0.007). Parasitized lambs of the more susceptible breed (S) showed anorexia [i.e., a decrease in intake of 13% compared with uninfected controls (P = 0.01)], whereas no significant reduction in food intake was observed in lambs of the more resistant breed (B). Reexposure to nematode infection of previously infected animals tended to result in renewed anorexia in S lambs but not in B lambs (P = 0.08) in a similar extent as during primary infection. Plasma leptin concentrations did not differ between ad libitum fed infected and control lambs but were greater in infected than in noninfected lambs at a similar level of food intake during both the primary (P = 0.02) and the secondary parasitic infection (P = 0.004) in both breeds. The results show that leptin may be involved in the response of lambs to infection but that it is unlikely that leptin alone is responsible for the parasite-induced anorexia in lambs. PMID- 18407985 TI - Porcine adipose triglyceride lipase complementary deoxyribonucleic acid clone, expression pattern, and regulation by resveratrol. AB - Adipose triglyceride lipase (ATGL) was recently identified and described as a major novel triglyceride lipase in animals. In this study, we aimed to study the tissue-specific and developmental expression pattern of porcine ATGL (pATGL) and the effect of resveratrol (RES) on expression of pATGL in vitro. The full-length cDNA sequence of pATGL was 1,958 bp (accession no. EF583921), with a 1,458-bp open reading frame encoding a 486-AA protein (the predicted molecular mass of 53.2 kDa, accession no. ABS58651). Comparison of the deduced AA sequence with the bovine, mouse, rat, dog, and human adipose triglyceride lipase showed 87, 84, 83, 81, and 80% similarity, respectively. Furthermore, the pATGL was highly expressed in porcine adipose tissue, to a lesser degree in kidney, heart, and muscle, and least but detectable in brain. In s.c. adipose tissue, pATGL mRNA was low at birth (1 kg of BW) and then increased, reaching a maximal value at 20 kg of BW (approximately 8 wk old; P < 0.01). In peritoneal and omental adipose tissue, the greatest expression of pATGL was observed at 40 kg of BW (approximately 12 wk old). In vitro, exposure of cultured adipocytes to 40 and 80 muM RES for 24 h increased the mRNA levels of pATGL by 95.3% (P < 0.05) and 146.8% (P < 0.01), respectively. Accordingly, lipid accumulation was decreased by 25.7% (P < 0.05) and 60.8% (P < 0.01), respectively. When treated with RES for 48 h, the mRNA levels of pATGL were increased by 104.1% (P < 0.05) and 163.1% (P < 0.01), respectively. As expected, lipid accumulation was decreased by 9.7% (P > 0.05) and 29.0% (P < 0.05), respectively. These results add to our understanding of the role of pATGL in adipose tissue development and as a potential target for regulating fat deposition and meat quality. PMID- 18407986 TI - Effects of dietary energy source and level and injection of tilmicosin phosphate on immune function in lipopolysaccharide-challenged beef steers. AB - Twenty-four Angus x Hereford crossbred steers (247 kg BW; SE = 2.4 kg) were used in a completely random design to evaluate the effect of energy source and level with or without antibiotic administration on measures of immune function. Steers were fed 1 of 3 dietary treatments: a 70% concentrate diet ad libitum (70AL), a 30% concentrate diet ad libitum (30AL), and a 70% concentrate diet offered in an amount calculated to provide NE(g) intake equal to the 30AL treatment (70RES). Half the steers in each dietary treatment received a s.c. injection of tilmicosin phosphate (ANTI; 1 mL/30 kg of BW); the other half received an equal volume of saline s.c. (SAL). Steers were offered the treatment diets for 28 d before and were administered the ANTI or SAL injections 2 d before indwelling catheters were placed in the jugular vein and 2.0 microg/kg of BW of Escherichia coli lipopolysaccharide (LPS) was administered i.v. Blood serum was collected at 30 min intervals from -2 to 6 h and at 8, 12, 24, 48, and 72 h relative to the LPS challenge. Increased energy intake (70AL) increased (P < or = 0.04) DMI, ADG, and rectal temperature (RT) after the challenge compared with the 70RES treatment. The 30AL treatment increased the maximum concentrations and area under the response curve of the proinflammatory cytokines (PIC) interferon-gamma, tumor necrosis factor-alpha, and IL-6 (P < or = 0.05) compared with the average of the 70AL and 70RES treatments. Decreased energy intake (70RES vs. 70AL) increased IL 6 (P < or = 0.003) but did not significantly increase interferon-gamma and tumor necrosis factor-alpha (P > or = 0.14) after LPS administration. Tilmicosin administration decreased the time to attain maximal RT (P = 0.01) by 1 h without altering the peak RT (P = 0.85), and tilmicosin interacted with energy intake to increase prechallenge PIC in 70RES vs. 70AL (P < or = 0.05). Results indicate that increased PIC response, presumably resulting from a combination of decreased energy intake and from direct effects of roughage, may be a mode of action for the slight decrease in morbidity that often occurs when newly received, stressed calves are fed roughage-based receiving diets. Tilmicosin phosphate might have immunomodulatory capacity beyond its direct effects on pathogenic bacteria, and these effects could interact with dietary energy intake in cattle. PMID- 18407987 TI - Evaluation of carcass characteristics of Bos indicus and tropically adapted Bos taurus breeds selected for postweaning weight. AB - Data from 9 studies were compiled to evaluate the effects of 20 yr of selection for postweaning weight (PWW) on carcass characteristics and meat quality in experimental herds of control Nellore (NeC) and selected Nellore (NeS), Caracu (CaS), Guzerah (GuS), and Gir (GiS) breeds. These studies were conducted with animals from a genetic selection program at the Experimental Station of Sertaozinho, Sao Paulo State, Brazil. After the performance test (168 d postweaning), bulls (n = 490) from the calf crops born between 1992 and 2000 were finished and slaughtered to evaluate carcass traits and meat quality. Treatments were different across studies. A meta-analysis was conducted with a random coefficients model in which herd was considered a fixed effect and treatments within year and year were considered as random effects. Either calculated maturity degree or initial BW was used interchangeably as the covariate, and least squares means were used in the multiple-comparison analysis. The CaS and NeS had heavier (P = 0.002) carcasses than the NeC and GiS; GuS were intermediate. The CaS had the longest carcass (P < 0.001) and heaviest spare ribs (P < 0.001), striploin (P < 0.001), and beef plate (P = 0.013). Although the body, carcass, and quarter weights of NeS were similar to those of CaS, NeS had more edible meat in the leg region than did CaS bulls. Selection for PWW increased rib-eye area in Nellore bulls. Selected Caracu had the lowest (most favorable) shear force values compared with the NeS (P = 0.003), NeC (P = 0.005), GuS (P = 0.003), and GiS (P = 0.008). Selection for PWW increased body, carcass, and meat retail weights in the Nellore without altering dressing percentage and body fat percentage. PMID- 18407989 TI - Comparison of protocols to synchronize estrus and ovulation in estrous-cycling and prepubertal beef heifers. AB - The objective of the experiment was to compare follicular dynamics, ovulatory response to GnRH, and synchrony of estrus and ovulation among estrous-cycling and prepubertal beef heifers synchronized with a controlled internal drug-release (CIDR)- based or GnRH-PGF(2alpha) (PG) protocol. Estrous-cycling beef heifers were randomly assigned to 1 of 4 treatments (C1, C2, C3, C4), and prepubertal beef heifers were randomly assigned to 1 of 2 treatments (P1, P2) by age and BW. Blood samples were taken 10 and 1 d before treatment to confirm estrous cyclicity status (progesterone > or =0.5 ng/mL estrous cycling). The CIDR Select (C1, n = 12; P1, n = 14)-treated heifers received a CIDR insert (1.38 g of progesterone) from d 0 to 14, GnRH (100 microg, i.m.) on d 23, and PG (25 mg, i.m.) on d 30. Select Synch + CIDR (C2, n = 12; P2, n = 11)-treated heifers received a CIDR insert and GnRH on d 23 and PG at CIDR removal on d 30. The CIDR-PG (C3, n = 12) treated heifers received a CIDR insert on d 23 and PG at CIDR removal on d 30. Select Synch (C4, n = 12)-treated heifers received GnRH on d 23 and PG on d 30. HeatWatch transmitters were fitted at CIDR removal (C1, C2, C3, P1, and P2) or at GnRH administration (C4) for estrus detection. Ultrasound was used to determine the response to GnRH and the timing of ovulation after estrus. Among the estrous cycling heifers, ovulatory response to GnRH and estrous response did not differ (P > 0.05). Among the prepubertal heifers, more (P = 0.02) P1 heifers responded to GnRH than P2 heifers, but estrous response did not differ (P > 0.05). Among the estrous-cycling heifers, variance for interval to estrus after PG was reduced (P < 0.05) for C1 compared with each of the other treatments, and C3 [corrected] was reduced (P < 0.05) compared with C2 [corrected] Variance for interval to ovulation after PG was reduced (P < 0.05) for C1 compared with each of the other treatments. Among the prepubertal heifers, there was no difference (P > 0.05) in variance for interval to estrus or ovulation. Results from C1 and P1 (T1) and C2 and P2 (T2) were combined to compare T1 and T2 among mixed groups of estrous cycling and prepubertal heifers. Response to GnRH was greater (P < 0.01; 81% T1 and 39% T2), and variances for interval to estrus and ovulation for T1 were reduced (P < 0.01) compared with T2. In summary, CIDR Select improved (P < 0.01) the synchrony of estrus and ovulation compared with Select Synch + CIDR. PMID- 18407990 TI - Cyst(e)ine imbalance and its effect on methionine precursor utilization in chicks. AB - Five 9- or 12-d chick growth bioassays were done in batteries using 2 Met deficient diets: a purified AA-based diet containing (by analysis, as-fed) 20.3% CP, 0.12% Met, and 0.05% cyst(e)ine; and an AA-fortified corn-peanut meal diet containing (by analysis, as-fed) 19.0% CP, 0.22% Met, and 0.23% cyst(e) ine. Feed grade DL-Met (dl-M; 99%) was compared with feed-grade DL-OH-Met, Ca (OH-M; 84%). When the purified diet was modified to contain 0.12% Met and 0.20% or greater cyst(e)ine, slope-ratio assays involving graded dosing of DL-M (0, 404, 808, and 1,212 mg of DL-M/kg) or isosulfurous levels of OH-M resulted in linear (P < 0.01) BW gain and G:F responses. Multiple linear regression analysis (BW gain vs. supplemental sulfur intake, R(2) = 0.98) resulted in a mean bioefficacy estimate of 78.1% for OH-M vs. DL-M (equivalent to 65.6% on a supplemental compound basis). In assay 3, the purified diet was modified to be equally deficient in Met and cyst(e)ine [i.e., 0.12% Met, 0.12% cyst(e)ine]. When this diet was supplemented with either 404 mg of DL-M/kg or 476 mg of OH-M/kg, BW gain and G:F responded (P < 0.01) markedly to either compound, and differences between DL-M and OH-M were not significant (P > 0.10). Assays 4 and 5 used the corn-peanut meal basal diet containing 0.22% total Met and 0.23% total cyst(e)ine. In both assays, addition of either 465 mg of DL-M/kg or 554 mg of OH-M/kg resulted in increased (P < 0.01) BW gain and G:F, regardless of dietary cyst(e)ine concentration. In the absence of excess cyst(e)ine, BW gain responses to DL-M and OH-M were similar, but when 0.10% excess cyst(e)ine was provided as L-cystine or feather meal, DL-M responses tended to exceed those of OH-M. Moreover, this small excess of dietary cyst(e)ine, regardless of source, depressed (P < 0.01) feed intake and BW gain when added to the basal diet. Overall, these results suggest that excess dietary cyst(e)ine, when included in Met-deficient diets, has the potential to be both anorexigenic and pernicious to OH-M utilization. PMID- 18407991 TI - The effect of residual feed intake classification on forage intake by grazing beef cows. AB - Although feed intake and efficiency differences in growing cattle of low and high residual feed intake (RFI) classification have been established, little is known about the difference in grazed forage intake between beef cows of known RFI classification. Two experiments were conducted using Hereford cows for which RFI had been determined as heifers using the GrowSafe 4000E feed intake system, after which heifers had been divided into thirds as low RFI, mid RFI, and high RFI. During Exp. 1, 2 replicates of low and high RFI cows (n = 7/replicate) in mid- to late-gestation were blocked to 1 of 4 non-endophyte-infected tall fescue paddocks (1.8 to 2.4 ha), which they grazed continuously for 84 d during summer. Using grazing exclosures, weekly rising plate meter readings, and forage harvests every 21 d, average forage DMI was calculated. Low and high RFI groups did not differ (P > 0.05) in BW change or BCS change over the trial (19.5 vs. 22.1 kg of BW gain and 0.11 vs. 0.10 BCS gain), but low RFI cows had a 21% numerically lower DMI than high RFI cows (12.4 vs. 15.6 kg/d; P = 0.23). The average area needed per paddock over the trial was similar for low and high RFI cows (1.71 vs. 1.82 ha; P = 0.35), and the average DM on offer over the trial was less for low RFI than for high RFI cows (4,215 vs. 4,376 kg; P = 0.06). During Exp. 2, 3 replicates of low and high RFI cows with their calves (n = 4 pair/replicate) strip-grazed stockpiled and early spring growth tall fescue paddocks (0.7 to 0.9 ha) for 60 d in late winter and early spring. Because of limiting forage availability and quality at trial initiation, cow-calf pairs were also fed 3.31 kg/pair of pelleted soyhulls daily. Pre- and post-grazed forage samples were harvested for 4 grazing periods, and forage growth was estimated using a growing degree days calculation and on-site weather station data. Performance did not differ (P > 0.05) between low and high RFI cows throughout the experiment (18.4 vs. 26.6 kg of BW gain and -0.04 vs. 0.15 BCS gain). Despite the utilization of forage offered being similar for low and high RFI cow-calf pairs (P > 0.05), low RFI cows and their calves had an 11% numerically lower DMI than high RFI pairs (12.5 vs. 14.1 kg/d; P = 0.12). We concluded that either no intake differences existed between low and high RFI cows or that current methodology and small animal numbers limited our ability to detect differences. PMID- 18407992 TI - Genetic parameters for early lamb survival and growth in Scottish Blackface sheep. AB - The objectives of this study were 1) to estimate the heritability of lamb survival and growth in the Scottish Blackface breed; 2) to examine the relationship between lamb survival and live BW; and 3) to investigate the possibility of using lamb survival in a breeding program for this breed. The data used for the analyses contained information about survival and live BW at different ages on 4,459 animals. The records were collected from 1988 to 2003 in a Scottish Blackface flock. Live BW was recorded every 4 wk from birth to 24 wk. Survival was defined either by perinatal or postnatal mortality (up to weaning at 12 wk), or as cumulative survival to 1, 4, 8, and 12 wk. The pedigree file comprised 1,416 dams and 178 sires. A sire model was used to estimate genetic parameters for binary survival traits. Heritabilities of BW traits, and phenotypic and genetic correlations between BW and between survival and BW were estimated by fitting an animal model. Further, correlations of survival with live BW were estimated by using a Markov chain Monte Carlo threshold model, implemented by Gibbs sampling. The heritability estimates for cumulative lamb survival declined from birth onward (from 0.33 to 0.08), and postnatal survival had a heritability of 0.01. The direct and maternal heritabilities for BW traits ranged from 0.08 to 0.26 and from 0.06 to 0.21, respectively, whereas the maternal environmental component was between 0.04 and 0.16. The genetic correlations between BW traits at different ages were high. The genetic and phenotypic correlations between survival and BW were always positive (ranging from 0.04 to 0.54), so there was no antagonism between these traits. Therefore, it is possible to simultaneously improve both survival and live BW in a breeding program for this breed. PMID- 18407993 TI - Forage systems for cow-calf production in the Appalachian region. AB - Small cow-calf operations are common in the Appalachian region. Tall fescue [Lolium arundinaceum (Schreb.) S. J. Darbyshire] is the dominant forage in these systems for direct grazing as well as for stockpiling. The present study was conducted from 2001 to 2005. A total of 108 Angus and Angus crossbred cows were allotted randomly to 6 forage systems and then to 3 replicates within each system. In brief, system 1 had a stocking rate of 0.91 ha/cow in a Middleburg 3 paddock (A, B, and C) system. System 2 was similar to system 1 except for a stocking rate of 0.71 ha/cow. A stocking rate of 0.71 ha/cow also was used in systems 3 through 6. All A paddocks had tall fescue, whereas B paddocks had tall fescue/white clover (Trifolium repens L.) except in system 6, which had tall fescue/lespedeza [Lespedeza cuneata (Dum. Cours.) G. Don]. System 3 evaluated a 2 paddock (A and B) rotational grazing system, and system 4 evaluated a 3-paddock (A, B, and C) rotational grazing system, with paddock C containing orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.). Systems 5 and 6 differed from system 2 in the areas of paddocks B and C as well as in the forage mixtures used. In paddock C, system 5 had switchgrass (Panicum virgatum L.) and system 6 had tall fescue and birdsfoot trefoil (Lotus corniculatus L.). System 1 had the greatest average herbage availability from weaning until breeding (P < 0.05) with the least amount of hay fed (P = 0.03) when compared with the remainder of the systems. Differences (P > 0.05) in percentage of ground cover were not detected among systems. There was no year x system interaction effect on the cow or calf performance variables evaluated and no treatment effect on cow performance variables. There was a treatment effect on calf performance variables. System 2 produced the greatest adjusted weaning weight, kilograms of calf weaned per hectare, and kilograms of calf per kilograms of cow at weaning (P < 0.05). Numerical ranking for total calf production per hectare from the greatest to least was system 2, 6, 3, 5, 4, and 1. Systems evaluated did not affect cow performance although differences in calf performance and overall productivity of the systems were observed. PMID- 18407994 TI - Effects of preshipping management on measures of stress and performance of beef steers during feedlot receiving. AB - Over 2 yr, a total of 96 steers (approximately 7 mo of age) were allocated to 1 of 4 weaning management strategies: 1) control: weaned on the day of shipping; 2) creep-fed: allowed free-choice access to concentrate before weaning and shipping; 3) preweaned: weaned and provided supplemental concentrate on pasture before shipping; and 4) early-weaned: weaned at 70 to 90 d of age and kept on pasture. On the day of shipping, steers were loaded together onto a commercial livestock trailer and transported 1,600 km over 24 h before being received into the feedlot. At the feedlot, steers were penned by treatment (4 pens/treatment) and provided access to free-choice hay and concentrate in separate feeding spaces. Samples of blood were collected on d 0, 1, 4, 8, 15, 22, and 29 relative to shipping. Steer performance was assessed over the receiving period, including DMI of hay and concentrate, ADG, and G:F. Predetermined contrasts included control vs. early-weaned, creep-fed vs. preweaned, and control vs. creep-fed and preweaned. Overall ADG was greater (P < 0.01) for early-weaned vs. control steers (1.39 vs. 0.88 kg). In wk 1, early-weaned steers consumed more concentrate and less hay compared with control steers (P < 0.03), and preweaned steers consumed more concentrate (P < 0.01) but a similar amount of hay (P = 0.75) compared with creep-fed steers. Average DMI was greater for preweaned compared with creep-fed steers (2.84 vs. 2.50% of BW; P = 0.01) and tended to be greater for early-weaned compared with control steers (2.76 vs. 2.50% of BW; P = 0.06). Feed efficiency of early-weaned steers was greater than that of control steers (G:F = 0.17 vs. 0.12; P < 0.01) but similar for preweaned compared with creep-fed steers (P = 0.72). Plasma ceruloplasmin concentrations were less (P < 0.05) in control vs. early weaned steers on d 0, but increased sharply after shipping and were greater in control vs. early-weaned steers on d 15 and 22 (P < 0.05). Creep-fed steers also experienced greater (P < 0.05) plasma ceruloplasmin concentrations than preweaned steers on d 29. These data suggest that early-weaned steers have improved performance in the feedlot compared with steers weaned directly before transport and feedlot entry. Differences in preshipping management appear to significantly affect measures of the acute phase protein response in steers. PMID- 18407995 TI - Effect of dietary soybean oil and soybean protein concentration on the concentration of digestible amino acids in soybean products fed to growing pigs. AB - An experiment was conducted to measure the effect of adding soybean oil to soybean meal (SBM) and soy protein concentrate (SPC) on apparent (AID) and standardized (SID) ileal digestibility of CP and AA by growing pigs. A second objective was to compare AID and SID of AA in a new high-protein variety of full fat soybeans (FFSB) to values obtained in other soybean products. Commercial sources of FFSB (FFSB-CV), SBM, and SPC, and of a new high-protein variety of FFSB (FFSB-HP) were used in the experiment. Four diets were prepared using each soybean product as the sole source of CP and AA in 1 diet. Two additional diets were formulated by adding soybean oil (7.55 and 7.35%, respectively) to the diets containing SBM and SPC. A nitrogen-free diet was also used to measure basal endogenous losses of CP and AA. The 2 sources of FFSB were extruded at 150 degrees C before being used in the experiment. Seven growing barrows (initial BW = 26.2 kg) were prepared with a T-cannula in the distal ileum and allotted to a 7 x 7 Latin square design. Ileal digesta were collected from the pigs on d 6 and 7 of each period. All digesta samples were lyophilized and analyzed for DM, CP, AA, and chromium, and values for AID and SID of CP and AA were calculated. The addition of oil improved (P < 0.05) the SID of most indispensable AA in SBM and SPC. The SID for 6 of the indispensable AA in FFSB-HP were greater (P < 0.05) than in FFSB-CV, and the SID for all indispensable AA except Met was greater (P < 0.05) in FFSB-HP than in SBM. However, the SID for most AA in FFSB-HP was similar to SBM with oil and SPC, but these values were lower (P < 0.05) than in SPC with oil. In conclusion, the addition of oil improved the SID of most AA in SBM and SPC fed to growing pigs, and the SID of AA in FFSB-HP were greater than in SBM and similar to the SID of AA in SBM with oil and in SPC. PMID- 18407996 TI - Assessment of the dynamics of microparasite infections in genetically homogeneous and heterogeneous populations using a stochastic epidemic model. AB - The aim of this paper was to explore the effect of genetic heterogeneity in host resistance to infection on the population-level risks and outcomes of epidemics. This was done using a stochastic epidemiological model in which the model parameters were assumed to be genetically controlled traits of the host. A finite locus model was explored, with a gene controlling the transmission coefficient (i.e., host susceptibility to infection) and a gene controlling the recovery period. Both genes were simulated to have 2 alleles with underlying additive or dominance inheritance and an independent assortment of alleles. The model was parameterized for a viral pig disease (transmissible gastroenteritis), and complete homogeneous mixing among genotypes was assumed. Mean population genotype dramatically affected epidemic outcomes, and subtle effects of heterogeneity on epidemic properties were also observed. Genetic variation in the transmission coefficient led to probabilities of epidemics occurring that were slightly greater than expected, but genetic variation in the recovery rate had no such effect. Epidemics were generally less severe in genetically heterogeneous populations than expected from the constituent subpopulations. Furthermore, the genotype of the initial infected animal had a marked effect on epidemic probabilities, particularly when genetic variation was for recovery rate. The results of this model provide useful information to determine the optimum population structures and to exploit genetic variation in resistance to infection. Applications of the proposed model in genetically heterogeneous populations for identifying practical disease management strategies are also discussed. PMID- 18407997 TI - Evaluation of three-ram cohort serving capacity tests as a substitute for individual serving capacity tests. AB - Alternatives to time-consuming, laborious individual serving capacity tests (ISCT) are needed to classify ram sexual behavior. The objective of study 1 was to evaluate the relationship between the first 3-ram cohort test (COSCT) scores and the mean of 5 ISCT scores. The objective of study 2 was to determine whether 1 or 2 additional COSCT improved the ability to predict ISCT scores. For study 1, rams (n = 69) were assigned to either a COSCT given before or after 9 ISCT. For study 2, rams (n = 127) were given 3 COSCT before or after 6 ISCT. For repeated COSCT, rams were initially grouped at random and subsequently rerandomized so that each ram was grouped with at least 1 different ram for each test. For both studies, the number of ejaculations from COSCT was compared with the mean number of ejaculations across the second through sixth ISCT. A threshold between high- and low-performing rams was defined in each analysis as the mean ISCT scores of sexually active rams. Rams with a mean number of ejaculations in ISCT greater than the threshold were classified as having high sexual activity, whereas rams below the threshold were classified as having low sexual activity. Rams with no ejaculations in ISCT were classified as sexually inactive. Data from studies 1 and 2 were used to evaluate the relationship between the first COSCT and the mean of 5 ISCT scores. Data from multiple COSCT were fit to various models to determine whether the ability to predict ISCT scores was improved with 1 or 2 additional COSCT. The best model for ISCT and COSCT was a piecewise linear regression model. The first COSCT correctly identified all sexually inactive rams in both studies. The first COSCT, however, also classified 56% of low sexually active rams and 18% of high sexually active rams as inactive. Rams had a 71% probability of high sexual activity in ISCT if they were classified as sexually active in the first COSCT. We conclude that a single COSCT is a reliable, albeit more conservative, and efficient alternative to a series of ISCT for characterizing sexual activity of rams. Multiple COSCT can provide some protection against culling rams with high sexual activity (i.e., approximately 50% less with 2 additional COSCT) and still retain most of the efficiency compared with ISCT. It is important to use high-performance rams for breeding because they will approximately double the number of ewes bred and lambs sired compared with low-performance rams if a large number of ewes need to be serviced daily. PMID- 18407998 TI - Crystal structure of human liver Delta4-3-ketosteroid 5beta-reductase (AKR1D1) and implications for substrate binding and catalysis. AB - AKR1D1 (steroid 5beta-reductase) reduces all Delta(4)-3-ketosteroids to form 5beta-dihydrosteroids, a first step in the clearance of steroid hormones and an essential step in the synthesis of all bile acids. The reduction of the carbon carbon double bond in an alpha,beta-unsaturated ketone by 5beta-reductase is a unique reaction in steroid enzymology because hydride transfer from NADPH to the beta-face of a Delta(4)-3-ketosteroid yields a cis-A/B-ring configuration with an approximately 90 degrees bend in steroid structure. Here, we report the first x ray crystal structure of a mammalian steroid hormone carbon-carbon double bond reductase, human Delta(4)-3-ketosteroid 5beta-reductase (AKR1D1), and its complexes with intact substrates. We have determined the structures of AKR1D1 complexes with NADP(+) at 1.79- and 1.35-A resolution (HEPES bound in the active site), NADP(+) and cortisone at 1.90-A resolution, NADP(+) and progesterone at 2.03-A resolution, and NADP(+) and testosterone at 1.62-A resolution. Complexes with cortisone and progesterone reveal productive substrate binding orientations based on the proximity of each steroid carbon-carbon double bond to the re-face of the nicotinamide ring of NADP(+). This orientation would permit 4-pro-(R) hydride transfer from NADPH. Each steroid carbonyl accepts hydrogen bonds from catalytic residues Tyr(58) and Glu(120). The Y58F and E120A mutants are devoid of activity, supporting a role for this dyad in the catalytic mechanism. Intriguingly, testosterone binds nonproductively, thereby rationalizing the substrate inhibition observed with this particular steroid. The locations of disease-linked mutations thought to be responsible for bile acid deficiency are also revealed. PMID- 18407999 TI - A p120 catenin isoform switch affects Rho activity, induces tumor cell invasion, and predicts metastatic disease. AB - p120 catenin is a cadherin-associated protein that regulates Rho GTPases and promotes the invasiveness of E-cadherin-deficient cancer cells. Multiple p120 isoforms are expressed in cells via alternative splicing, and all of them are essential for HGF signaling to Rac1. However, only full-length p120 (isoform 1) promotes invasiveness. This selective ability of p120 isoform 1 is mediated by reduced RhoA activity, both under basal conditions and following HGF treatment. All p120 isoforms can bind RhoA in vitro, via a central RhoA binding site. However, only the cooperative binding of RhoA to the central p120 domain and to the alternatively spliced p120 N terminus stabilizes RhoA binding and inhibits RhoA activity. Consistent with this, increased expression of p120 isoform 1, when compared with other p120 isoforms, is predictive of renal tumor micrometastasis and systemic progression, following nephrectomy. Furthermore, ectopic expression of the RhoA-binding, N-terminal domain of p120 is sufficient to block the ability of p120 isoform 1 to inhibit RhoA and to promote invasiveness. The data indicate that the increased expression of p120 isoform 1 during tumor progression contributes to the invasive phenotype of cadherin-deficient carcinomas and that the N-terminal domain of p120 is a valid therapeutic target. PMID- 18408000 TI - Surface exposure of the HIV-1 env cytoplasmic tail LLP2 domain during the membrane fusion process: interaction with gp41 fusion core. AB - HIV-1 gp41 cytoplasmic tail (CT) is highly conserved among HIV-1 isolates, particularly the region designated lentivirus lytic peptide (LLP1-2), which includes two alpha-helical domains LLP1 and LLP2. Although the gp41 CT is recognized as a modulator of viral fusogenicity, little is known about the regulatory mechanism of this region in the viral fusion process. Here we report that anti-LLP1-2 and anti-LLP2 antibodies (IgG) inhibited HIV-1 Env-mediated cell fusion and bound to the interface between effector and target cells at a suboptimal temperature (31.5 degrees C), which slows down the fusion process and prolongs the fusion intermediate state. This suggests that LLP1-2, especially the LLP2 region located inside the viral membrane, is transiently exposed on the membrane surface during the fusion process. Synthetic LLP2 peptide could bind to the gp41 six-helix bundle core with high binding affinity. These results suggest that the gp41 CT may interact with the gp41 core, via the surface-exposed LLP2 domain, to regulate Env-mediated membrane fusion. PMID- 18408001 TI - Phosphorylation of CCAAT/enhancer-binding protein alpha regulates GLUT4 expression and glucose transport in adipocytes. AB - The transcription factor CCAAT/enhancer-binding protein alpha (C/EBPalpha) is required during adipogenesis for development of insulin-stimulated glucose uptake. Modes for regulating this function of C/EBPalpha have yet to be determined. Phosphorylation of C/EBPalpha on Ser-21 has been implicated in the regulation of granulopoiesis and hepatic gene expression. To explore the role of Ser-21 phosphorylation on C/EBPalpha function during adipogenesis, we developed constructs in which Ser-21 was mutated to alanine (S21A) to model dephosphorylation. In two cell culture models deficient in endogenous C/EBPalpha, enforced expression of S21A-C/EBPalpha resulted in normal lipid accumulation and expression of many adipogenic markers. However, S21A-C/EBPalpha had impaired ability to activate the Glut4 promoter specifically, and S21A-C/EBPalpha expression resulted in diminished GLUT4 and adiponectin expression, as well as reduced insulin-stimulated glucose uptake. No defects in insulin signaling or GLUT4 vesicle trafficking were identified with S21A-C/EBPalpha expression, and when exogenous GLUT4 expression was enforced to normalize expression in S21A C/EBPalpha cells, insulin-responsive glucose transport was reconstituted, suggesting that the primary defect was a deficit in GLUT4 levels. Mice in which endogenous C/EBPalpha was replaced with S21A-C/EBPalpha displayed reduced GLUT4 and adiponectin protein expression in epididymal adipose tissue and increased blood glucose compared with wild-type littermates. These results suggest that phosphorylation of C/EBPalpha on Ser-21 may regulate adipocyte gene expression and whole body glucose homeostasis. PMID- 18408002 TI - Platyhelminth mitochondrial and cytosolic redox homeostasis is controlled by a single thioredoxin glutathione reductase and dependent on selenium and glutathione. AB - Platyhelminth parasites are a major health problem in developing countries. In contrast to their mammalian hosts, platyhelminth thiol-disulfide redox homeostasis relies on linked thioredoxin-glutathione systems, which are fully dependent on thioredoxin-glutathione reductase (TGR), a promising drug target. TGR is a homodimeric enzyme comprising a glutaredoxin domain and thioredoxin reductase (TR) domains with a C-terminal redox center containing selenocysteine (Sec). In this study, we demonstrate the existence of functional linked thioredoxin-glutathione systems in the cytosolic and mitochondrial compartments of Echinococcus granulosus, the platyhelminth responsible for hydatid disease. The glutathione reductase (GR) activity of TGR exhibited hysteretic behavior regulated by the [GSSG]/[GSH] ratio. This behavior was associated with glutathionylation by GSSG and abolished by deglutathionylation. The K(m) and k(cat) values for mitochondrial and cytosolic thioredoxins (9.5 microm and 131 s( 1), 34 microm and 197 s(-1), respectively) were higher than those reported for mammalian TRs. Analysis of TGR mutants revealed that the glutaredoxin domain is required for the GR activity but did not affect the TR activity. In contrast, both GR and TR activities were dependent on the Sec-containing redox center. The activity loss caused by the Sec-to-Cys mutation could be partially compensated by a Cys-to-Sec mutation of the neighboring residue, indicating that Sec can support catalysis at this alternative position. Consistent with the essential role of TGR in redox control, 2.5 microm auranofin, a known TGR inhibitor, killed larval worms in vitro. These studies establish the selenium- and glutathione-dependent regulation of cytosolic and mitochondrial redox homeostasis through a single TGR enzyme in platyhelminths. PMID- 18408003 TI - Smooth muscle myosin phosphorylated at single head shows sustained mechanical activity. AB - Smooth muscle contraction is regulated by the phosphorylation of myosin. It is well known that tonic smooth muscles can maintain force with low energy consumption (latch state); however, the molecular mechanism underlying this phenomenon is unresolved. Here we show that single-head phosphorylated smooth myosin (SHPMII) exhibits fast ( approximately 24 s(-1)) and slow prolonged ( approximately 1 s(-1)) actin interactions, whereas double-head phosphorylated myosin (DHPMII) predominantly exhibits the fast ( approximately 29 s(-1)) interaction, suggesting that the phosphorylated head of SHPMII is mechanically as active as that of DHPMII. Both the fast and the slow actin interactions of SHPMII support the positive net mechanical displacement of actin. The actin translocating velocity of SHPMII was much slower than that of DHPMII, which is consistent with the slow actin interaction of SHPMII. We propose that the "latch state" can be explained by the motor characteristics of SHPMII that is present during the sustained phase of contraction. PMID- 18408004 TI - Anaerobic sulfatase-maturating enzymes, first dual substrate radical S adenosylmethionine enzymes. AB - Sulfatases are a major group of enzymes involved in many critical physiological processes as reflected by their broad distribution in all three domains of life. This class of hydrolases is unique in requiring an essential post-translational modification of a critical active-site cysteine or serine residue to C(alpha) formylglycine. This modification is catalyzed by at least three nonhomologous enzymatic systems in bacteria. Each enzymatic system is currently considered to be dedicated to the modification of either cysteine or serine residues encoded in the sulfatase-active site and has been accordingly categorized as Cys-type and Ser-type sulfatase-maturating enzymes. We report here the first detailed characterization of two bacterial anaerobic sulfatase-maturating enzymes (anSMEs) that are physiologically responsible for either Cys-type or Ser-type sulfatase maturation. The activity of both enzymes was investigated in vivo and in vitro using synthetic substrates and the successful purification of both enzymes facilitated the first biochemical and spectroscopic characterization of this class of enzyme. We demonstrate that reconstituted anSMEs are radical S-adenosyl l-methionine enzymes containing a redox active [4Fe-4S](2+,+) cluster that initiates the radical reaction by binding and reductively cleaving S-adenosyl-l methionine to yield 5 '-deoxyadenosine and methionine. Surprisingly, our results show that anSMEs are dual substrate enzymes able to oxidize both cysteine and serine residues to C(alpha)-formylglycine. Taken together, the results support a radical modification mechanism that is initiated by hydrogen abstraction from a serine or cysteine residue located in an appropriate target sequence. PMID- 18408005 TI - The beta-arrestin-2 scaffold protein promotes c-Jun N-terminal kinase-3 activation by binding to its nonconserved N terminus. AB - The c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) signaling pathway mediates stress responses in cells. JNK activity is regulated by a protein kinase cascade consisting of a MAPK kinase (MKK) and a MAPK kinase kinase (MAPKKK). beta-Arrestin-2 acts as a scaffold by directly binding to the JNK3 isoform and also by recruiting MKK4 and the MAPKKK apoptosis-signaling kinase-1 (ASK1). In this study, we demonstrate by co-precipitation that the extended N-terminal region of JNK3 mediates binding to the C terminus of beta arrestin-2 and that the N terminus of JNK3 is required for its activation via beta-arrestin-2. We have used site-specific mutagenesis to identify key residues within the N terminus of JNK3 that are essential for binding and demonstrate that this region represents an independent beta-arrestin-2 binding motif that can be fused to other MAPKs and permit their recruitment to the scaffold complex. In addition, we demonstrate that JNK3 recruits MKK4 to the beta-arrestin-2 scaffold complex by binding to the MAPK docking domain (D-domain) located within the N terminus of MKK4. These findings uncover molecular determinants of beta-arrestin 2 scaffold complex assembly and assign a previously unrecognized role for the unique extended N terminus of JNK3. PMID- 18408006 TI - CLEC9A is a novel activation C-type lectin-like receptor expressed on BDCA3+ dendritic cells and a subset of monocytes. AB - We describe here the first characterization of CLEC9A, a group V C-type lectin like receptor located in the "Dectin-1 cluster" of related receptors, which are encoded within the natural killer (NK)-gene complex. Expression of human CLEC9A is highly restricted in peripheral blood, being detected only on BDCA3(+) dendritic cells and on a small subset of CD14(+)CD16(-) monocytes. CLEC9A is expressed at the cell surface as a glycosylated dimer and can mediate endocytosis, but not phagocytosis. CLEC9A possesses a cytoplasmic immunoreceptor tyrosine-based activation-like motif that can recruit Syk kinase, and we demonstrate, using receptor chimeras, that this receptor can induce proinflammatory cytokine production. These data indicate that CLEC9A functions as an activation receptor. PMID- 18408007 TI - FoxM1c counteracts oxidative stress-induced senescence and stimulates Bmi-1 expression. AB - The Forkhead box transcription factor FoxM1 is expressed in proliferating cells. When it was depleted in mice and cell lines, cell cycle defects and chromosomal instability resulted. Premature senescence was observed in embryonic fibroblasts derived from FoxM1 knock-out mice, but the underlying cause has remained unclear. To investigate whether FoxM1 can protect cells against stress-induced premature senescence, we established NIH3T3 lines with doxycycline-inducible overexpression of FoxM1c. Treatment of these lines with sublethal doses (20 and 100 microm) of H(2)O(2) induced senescence with senescence-associated beta-galactosidase expression and elevated levels of p53 and p21. Induction of FoxM1c expression markedly suppressed senescence and expression of p53 and p21. Consistent with down-regulation of the p19(Arf)-p53 pathway, p19(Arf) levels decreased while expression of the Polycomb group protein Bmi-1 was induced. That Bmi-1 is a downstream target of FoxM1c was further supported by the dose-dependent induction of Bmi-1 by FoxM1c at both the protein and mRNA levels, and FoxM1 and Bmi-1 reached maximal levels in cells at the G(2)/M phase. Depletion of FoxM1 by RNA interference decreased Bmi-1 expression. Using Bmi-1 promoter reporters with wild type and mutated c-Myc binding sites and short hairpin RNAs targeting c-Myc, we further demonstrated that FoxM1c activated Bmi-1 expression via c-Myc, which was recently reported to be regulated by FoxM1c. Our results reveal a functional link between FoxM1c, c-Myc, and Bmi-1, which are major regulators of tumorigenesis. This link has important implications for the regulation of cell proliferation and senescence by FoxM1 and Bmi-1. PMID- 18408008 TI - The Caenorhabditis elegans AMP-activated protein kinase AAK-2 is phosphorylated by LKB1 and is required for resistance to oxidative stress and for normal motility and foraging behavior. AB - AAK-2 is one of two alpha isoforms of the AMP-activated protein kinase in Caenorhabditis elegans and is involved in life span maintenance, stress responses, and germ cell cycle arrest upon dauer entry. We found that AAK-2 was phosphorylated at threonine 243 in response to paraquat treatment and that this phosphorylation depends on PAR-4, the C. elegans LKB1 homologue. Both aak-2 mutation and par-4 knockdown increased the sensitivity of C. elegans worms to paraquat, and the double deficiency did not further increase sensitivity, indicating that aak-2 and par-4 act in a linear pathway. Both mutations also slowed body bending during locomotion and failed to reduce head oscillation in response to anterior touch. Consistent with this abnormal motility and behavioral response, expression of the AAK-2::green fluorescent protein fusion protein was observed in the ventral cord, some neurons, body wall muscle, pharynx, vulva, somatic gonad, and excretory cell. Our study suggests that AMPK can influence the behavior of C. elegans worms in addition to its well known function in metabolic control. PMID- 18408009 TI - USP11 stabilizes HPV-16E7 and further modulates the E7 biological activity. AB - HPV-16E7 is a major transforming protein, which has been implicated in the development of cervical cancer. The stability of E7 is thus important to ensure its fully functional status. Using the yeast two-hybrid system, we found that USP11 (ubiquitin-specific protease 11), a member of a protein family that cleaves polyubiquitin chains and/or ubiquitin precursors, interacts and forms a specific complex with HPV-16E7. Our results indicate that the USP11 can greatly increase the steady state level of HPV-16E7 by reducing ubiquitination and attenuating E7 degradation. In contrast, a catalytically inactive mutant of USP11 abolished the deubiquitinating ability and returned E7 to a normal rate of degradation. Moreover, USP11 not only protected E7 from ubiquitination but also influenced E7 function as a modulator of cell growth status. These results suggest that USP11 plays an important role in regulating the levels of E7 protein and subsequently affects the biological function of E7 as well as its contribution to cell transformation by HPV-16E7. PMID- 18408010 TI - A mutation in telethonin alters Nav1.5 function. AB - Excitable cells express a variety of ion channels that allow rapid exchange of ions with the extracellular space. Opening of Na(+) channels in excitable cells results in influx of Na(+) and cellular depolarization. The function of Na(v)1.5, an Na(+) channel expressed in the heart, brain, and gastrointestinal tract, is altered by interacting proteins. The pore-forming alpha-subunit of this channel is encoded by SCN5A. Genetic perturbations in SCN5A cause type 3 long QT syndrome and type 1 Brugada syndrome, two distinct heritable arrhythmia syndromes. Mutations in SCN5A are also associated with increased prevalence of gastrointestinal symptoms, suggesting that the Na(+) channel plays a role in normal gastrointestinal physiology and that alterations in its function may cause disease. We collected blood from patients with intestinal pseudo-obstruction (a disease associated with abnormal motility in the gut) and screened for mutations in SCN5A and ion channel-interacting proteins. A 42-year-old male patient was found to have a mutation in the gene TCAP, encoding for the small protein telethonin. Telethonin was found to be expressed in the human gastrointestinal smooth muscle, co-localized with Na(v)1.5, and co-immunoprecipitated with sodium channels. Expression of mutated telethonin, when co-expressed with SCN5A in HEK 293 cells, altered steady state activation kinetics of SCN5A, resulting in a doubling of the window current. These results suggest a new role for telethonin, namely that telethonin is a sodium channel-interacting protein. Also, mutations in telethonin can alter Na(v)1.5 kinetics and may play a role in intestinal pseudo-obstruction. PMID- 18408011 TI - MicroRNA-mediated systemic down-regulation of copper protein expression in response to low copper availability in Arabidopsis. AB - In plants, copper is an essential micronutrient required for photosynthesis. Two of the most abundant copper proteins, plastocyanin and copper/zinc superoxide dismutase, are found in chloroplasts. Whereas plastocyanin is essential for photo autotrophic growth, copper/zinc superoxide dismutase is dispensable and in plastids can be replaced by an iron superoxide dismutase when copper is limiting. The down-regulation of copper/zinc superoxide dismutase expression in response to low copper involves a microRNA, miR398. Interestingly, in Arabidopsis and other plants, three additional microRNA families, miR397, miR408, and miR857, are predicted to target the transcripts for the copper protein plantacyanin and members of the laccase copper protein family. We confirmed the predicted targets of miR397, miR408, and miR857 experimentally by cleavage site analysis. To study the spatial expression pattern of these microRNAs and the effect of copper on their expression, we analyzed Arabidopsis grown hydroponically on different copper regimes. On low amounts of copper the plants accumulated miR397, miR408, and miR857. The microRNA expression pattern was negatively correlated with the accumulation of transcripts for plantacyanin and laccases. Furthermore, the expression of other laccases that are not predicted targets for known microRNAs was similarly regulated in response to copper. For some of these laccases, the regulation was disrupted in a microRNA maturation mutant (hen1-1), suggesting the presence of other copper-regulated microRNAs. Thus, in Arabidopsis, microRNA mediated down-regulation is a general mechanism to regulate nonessential copper proteins. We propose that this mechanism allows plants to save copper for the most essential functions during limited copper supply. PMID- 18408012 TI - Cdk5-mediated phosphorylation of c-Myc on Ser-62 is essential in transcriptional activation of cyclin B1 by cyclin G1. AB - It has been reported previously that cyclin G1 enables cells to overcome radiation-induced G(2) arrest and increased cell death and that these effects are mediated by transcriptional activation of cyclin B1. In this study, we further investigated the mechanism by which cyclin G1 transcriptionally activates cyclin B1. Deletion or point mutations within the cyclin B1 promoter region revealed that the c-Myc binding site (E-box) is necessary for cyclin G1-mediated transcriptional activation of cyclin B1 to occur. In addition, the kinase activity of Cdk5 was increased by cyclin G1 overexpression, and Cdk5 directly phosphorylated c-Myc on Ser-62. Furthermore, cyclin G1 mediated increased radiosensitivity, and radiation-induced M phase arrest was attenuated when RNA interference of Cdk5 was treated. Taken together, the results of this study indicate that Cdk5 activation in cells that overexpress cyclin G1 leads to c-Myc phosphorylation on Ser-62, which is responsible for cyclin G1-mediated transcriptional activation of cyclin B1. PMID- 18408013 TI - Structure and dynamics of human apolipoprotein CIII. AB - Human apolipoprotein CIII (apoCIII) is a surface component of chylomicrons, very low density lipoproteins, and high density lipoproteins. ApoCIII inhibits lipoprotein lipase as well as binding of lipoproteins to cell surface heparan sulfate proteoglycans and receptors. High levels of apoCIII are often correlated with elevated levels of blood lipids (hypertriglyceridemia). Here, we report the three-dimensional NMR structure and dynamics of human apo-CIII in complex with SDS micelles, mimicking its natural lipid-bound state. Thanks to residual dipolar coupling data, the first detailed view is obtained of the structure and dynamics of an intact apolipoprotein in its lipid-bound state. ApoCIII wraps around the micelle surface as a necklace of six approximately 10-residue amphipathic helices, which are curved and connected via semiflexible hinges. Three positively charged (Lys) residues line the polar faces of helices 1 and 2. Interestingly, their three-dimensional conformation is similar to that of the low density lipoprotein receptor binding motifs of apoE/B and the receptor-associated protein. At the C-terminal side of apoCIII, an array of negatively charged residues lines the polar faces of helices 4 and 5 and the adjacent flexible loop. Sequence comparison shows that this asymmetric charge distribution along the solvent-exposed face of apoCIII as well as other structural features are conserved among mammals. This structure provides a template for exploration of molecular mechanisms by which human apoCIII inhibits lipoprotein lipase and receptor binding. PMID- 18408015 TI - Protein kinase Cepsilon binds peripherin and induces its aggregation, which is accompanied by apoptosis of neuroblastoma cells. AB - A hallmark of the afflicted nervous tissue in amyotrophic lateral sclerosis is the presence of protein aggregates, which to a large extent contain the intermediate filament protein peripherin. Here we show that activation of protein kinase C (PKC) or overexpression of PKCepsilon induces the aggregation of peripherin in cultured neuroblastoma cells with elevated amounts of peripherin. The formation of aggregates was coupled to an increased apoptosis, suggesting a functional link between these events. Both induction of aggregates and apoptosis were suppressed in cells that had been transfected with small interfering RNAs targeting PKCepsilon. PKCepsilon and peripherin associate as shown by co immunoprecipitation, and the interaction is dependent on and mediated by the C1b domain of PKCepsilon. The interaction was specific for PKCepsilon since corresponding structures from other isoforms did not co-precipitate peripherin, with the exception for PKCeta and -, which pulled down minute amounts. PKCepsilon interacts with vimentin through the same structures but does not induce its aggregation. When the PKCepsilon C1b domain is expressed in neuroblastoma cells together with peripherin, both phorbol ester-induced peripherin aggregation and apoptosis are abolished, supporting a model in which PKCepsilon through its interaction with peripherin facilitates its aggregation and subsequent cell death. These events may be prevented by expressing molecules that bind peripherin at the same site as PKCepsilon. PMID- 18408014 TI - SUMO conjugation to the matrix attachment region-binding protein, special AT-rich sequence-binding protein-1 (SATB1), targets SATB1 to promyelocytic nuclear bodies where it undergoes caspase cleavage. AB - SATB1 (special AT-rich sequence-binding protein-1) provides a key link between DNA loop organization, chromatin modification/remodeling, and association of transcription factors at matrix attachment regions (MARs). To investigate the role of SATB1 in cellular events, we performed a yeast two-hybrid screen that identified SUMO-1, Ubc9, and protein inhibitor of activated STAT (PIAS) family members as SATB1 interaction partners. These proteins, working in concert, enhanced SUMO conjugation to lysine-744 of SATB1. Overexpression of SUMO or PIAS in Jurkat cells, which express high levels of endogenous SATB1, exhibited enhanced caspase cleavage of this MAR-associating protein. Sumoylation-deficient SATB1 (SATB1(K744R)) failed to display the characteristic caspase cleavage pattern; however, fusion of SUMO in-frame to SATB1(K744R) restored cleavage. A SUMO-independent interaction of inactive caspase-6 and SATB1 was noted. A subset of total cellular SATB1 localized into promyelocytic leukemia nuclear bodies where enhanced SATB1 cleavage was detected subsequent to caspase activation. These results reveal a novel sumoylation-directed caspase cleavage of this key regulatory molecule. The role of regulated proteolysis of SATB1 may be to control transcription in immune cells during normal cell functions or to assist in efficient and rapid clearance of nonfunctional or potentially damaging immune cells. PMID- 18408016 TI - Identification of the pore-lining residues of the BM2 ion channel protein of influenza B virus. AB - The influenza B virus BM2 proton-selective ion channel is essential for virus uncoating, a process that occurs in the acidic environment of the endosome. The BM2 channel causes acidification of the interior of the virus particle, which results in dissociation of the viral membrane protein from the ribonucleo-protein core. The BM2 protein is similar to the A/M2 protein ion channel of influenza A virus (A/M2) in that it contains an HXXXW motif. Unlike the A/M2 protein, the BM2 protein is not inhibited by the antiviral drug amantadine. We used mutagenesis to ascertain the pore-lining residues of the BM2 ion channel. The specific activity (relative to wild type), reversal voltage, and susceptibility to modification by (2-aminoethyl)-methane thiosulfonate and N-ethylmaleimide of cysteine mutant proteins were measured in oocytes. It was found that mutation of transmembrane domain residues Ser(9), Ser(12), Phe(13), Ser(16), His(19), and Trp(23) to cysteine were most disruptive for ion channel function. These cysteine mutants were also most susceptible to (2-aminoethyl)-methane thiosulfonate and N ethylmaleimide modification. Furthermore, considerable amounts of dimer were formed in the absence of oxidative reagents when cysteine was introduced at positions Ser(9), Ser(12), Ser(16), or Trp(23). Based on these experimental data, a BM2 transmembrane domain model is proposed. The presence of polar residues in the pore is a probable explanation for the amantadine insensitivity of the BM2 protein and suggests that related but more polar compounds might serve as useful inhibitors of the protein. PMID- 18408017 TI - Estrogen addition to progesterone for luteal phase support in cycles stimulated with GnRH analogues and gonadotrophins for IVF: a systematic review and meta analysis. AB - BACKGROUND: The purpose of the present systematic review and meta-analysis was to examine whether the probability of pregnancy is increased by adding estrogen to progesterone for luteal phase support in patients treated by in vitro fertilization (IVF). METHODS: A literature search covering MEDLINE, EMBASE, CENTRAL, meeting proceedings and reference lists of published articles was performed to identify relevant RCTs. Data were extracted for meta-analysis yielding pooled relative risks (RR) and 95% confidence intervals (CI). Sensitivity analyses by including studies with pseudo-randomization or unclear method of randomization were also performed (n=1141 patients in total). RESULTS: Four RCTs (n=587 patients) were eligible for inclusion. No statistically significant differences were present between patients who received a combination of progesterone and estrogen for luteal support when compared with those who received only progesterone, in terms of positive hCG rate (RR: 1.02, 95% CI: 0.87 1.19), clinical pregnancy rate (RR: 0.94, 95% CI: 0.78-1.13) and live birth rate (RR: 0.96, 95% CI: 0.77-1.21) per woman randomized. These results did not materially differ in the sensitivity analyses performed. CONCLUSIONS: The currently available evidence suggests that the addition of estrogen to progesterone for luteal phase support does not increase the probability of pregnancy in IVF. However, there is an obvious need for further RCTs that will assess, with more confidence, the effect of estrogen addition to progesterone during the luteal phase on the probability of pregnancy. PMID- 18408018 TI - Fas receptor is not present on ejaculated human sperm. AB - BACKGROUND: Apoptosis appears to have an essential role in the control of testis germ cell number and Fas expression has been reported in apoptotic spermatocytes and spermatids. We investigated if Fas (CD95) was present on ejaculated human sperm and any relationship between Fas on sperm and the apoptotic marker Syto16. METHODS: Semen samples from 77 male partners of infertile couples were evaluated. Each sample was analysed both before and after semen preparation by conventional microscopical procedures and by flow cytometry (FC). A multiparameter FC analysis to assess simultaneously sperm concentration, sperm viability, sperm apoptosis, CD45 positive (leukocyte) and CD95 (Fas) positive cell concentration was carried out. A further 10 samples were studied by indirect immunofluorescence to confirm results. RESULTS: The mean concentration of CD95 positive cells was very low (<1%), with no significant difference between normozoospermic and non normozoospermic men. There was no correlation between apoptotic sperm and CD95 positive cell concentration. A linear correlation was found between CD95 positive cell and leukocyte (CD45 positive) concentration (r = 0.9946, P < 0.0001). CD95 mean fluorescence intensity of leukocytes was 10-fold greater than that of sperm and of isotypic control. Both incubation with activating anti-Fas antibody and betulinic acid induced apoptosis in leukocytes. Incubation with betulinic acid, but not with activating anti-Fas antibody, induced apoptosis in sperm. Pre incubation with neutralizing anti-Fas antibody suppressed CD95 expression on leukocytes, whereas it did not change sperm CD95 peak fluorescence. CONCLUSIONS: There is no detectable quantity of Fas on human ejaculated sperm. PMID- 18408019 TI - Myeloid ecotropic viral integration site 1 (MEIS) 1 involvement in embryonic implantation. AB - BACKGROUND: The HOXA10 homeobox gene controls embryonic uterine development and adult endometrial receptivity. The three-amino-acid loop extension (TALE) family homeobox genes like myeloid ecotropic viral integration site 1 (MEIS) provide enhanced target gene activation and specificity in HOX-regulated cellular processes by acting as HOX cofactors. METHODS AND RESULTS: Analysis of an Affymetrix data set in the public domain showed high expression of MEIS1 in human endometrium. MEIS1 expression was confirmed during the human menstrual cycle by RT-PCR and in situ hybridization and was increased during the secretory compared with proliferative phase of the cycle (P = 0.0001), the time of implantation. To assess the importance of maternal Meis1 expression in a mouse model, the uteri of Day 2 pregnant mice were injected with Meis1 over-expression or small interfering RNA (siRNA) constructs. Blocking Meis1 expression by siRNA before implantation significantly reduced average implantation rates (P = 0.00001). Increased or decreased Meis1 expression significantly increased or decreased the expression of integrin beta3, a transcriptional target of HOXA10 and an important factor in early embryo-endometrium interactions (P = 0.006). Manipulating Meis1 expression before implantation also dramatically affected the number of pinopodes, uterine endometrial epithelial projections that develop at the time of endometrial receptivity. CONCLUSIONS: The results suggest that in mouse, meis1 contributes to regulating endometrial development during the menstrual cycle and establishing the conditions necessary for implantation. PMID- 18408020 TI - Gene expression analysis of interferon-beta treatment in multiple sclerosis. AB - Treatment with interferon-beta (IFN-beta) induces the expression of hundreds of genes in blood mononuclear cells, and the expression of several genes has been proposed as a marker of the effect of treatment with IFN-beta. However, to date no molecules have been identified that are stably induced by treatment with IFN beta. We use DNA microarrays to study gene expression in 10 multiple sclerosis (MS) patients who began de novo treatment with IFN-beta. After the first injection of IFN-beta, the expression of 74 out of 3428 genes changed at least two-fold and statistically significantly (after Bonferroni correction). In contrast, we observed no persisting effects of IFN-beta on gene expression. Among the most strongly induced genes was MXA, which has been used in previous biomarker studies in MS. In addition, the study identified the induction of LGALS9 and TCIR1G, involved in negative regulation of T helper type I immunity and T-cell activation, as novel effects of IFN-beta therapy in MS. PMID- 18408021 TI - The role of iron dysregulation in the pathogenesis of multiple sclerosis: an Egyptian study. AB - BACKGROUND: Iron is essential for virtually all types of cells and organisms. The significance of iron for brain function is reflected by the presence of receptors for transferrin on brain capillary endothelial cells. Iron imbalance is associated with proinflammatory cytokines and oxidative stress, which have been implicated in the pathogenesis of multiple sclerosis (MS). Transferrin receptor (TfR) is the major mediator of iron uptake. Its expression is increased to facilitate iron entrance into the cell. The increased serum level of soluble transferrin receptor (sTfR) may indicate an abnormal intracellular distribution of iron and a decrease in the cytoplasmic compartment. OBJECTIVE: Our objective is to assess the possible role of iron metabolism dysfunction in the pathogenesis of MS. METHODS: Thirty subjects were selected from the Neurology Department of Kasr El-Aini hospital, Cairo University: 20 MS patients, where nine patients were relapsing and progressive (secondary progressive (SP) of which six were secondary progressive active (SP-A) and three were secondary progressive stable (SP-S)), seven were relapsing-remitting active (RR-A) and four were primary progressive (PP); and 10 control subjects matched in age and sex. Each patient was subjected to a thorough general medical and neurological examination, Kurtzke MS rating scales, laboratory assessment, neuro-imaging, evoked potentials and quantitative determination of the indices of iron metabolism, such as serum iron and sTfR. RESULTS: The serum level of sTfR was significantly higher in our MS patients compared with the control group (p = 0.0001). The levels were significantly higher in SP-A (p = 0.001), SP-S (p = 0.01), RR-A (p = 0.0001) and PP (p = 0.003) patients than in controls. Iron values were within normal limits in all patients. The increased serum sTfR level in non-anemic MS patients with active disease reflects the increased iron turnover. The elevation of sTfR levels in stable patients may indicate active inflammation with ongoing oxidative damage that is not detectable by history or examination. CONCLUSIONS: Iron overload and upregulation of iron-handling proteins, such as TfR, in the MS brain can contribute to pathogenesis of Multiple Sclerosis and iron imbalance is associated with a pro-oxidative stress and a proinflammatory environment, this suggest that iron could be a target for MS therapy to improve neuronal iron metabolism. PMID- 18408022 TI - The Aeromonas hydrophila wb*O34 gene cluster: genetics and temperature regulation. AB - The Aeromonas hydrophila wb*(O34) gene cluster of strain AH-3 (serotype O34) was cloned and sequenced. This cluster contains genes necessary for the production of O34-antigen lipopolysaccharide (LPS) in A. hydrophila. We determined, using either mutation or sequence homology, roles for the majority of genes in the cluster by using the chemical O34-antigen LPS structure obtained for strain AH-3. The O34-antigen LPS export system has been shown to be a Wzy-dependent pathway typical of heteropolysaccharide pathways. Furthermore, the production of A. hydrophila O34-antigen LPS in Escherichia coli K-12 strains is dependent on incorporation of the Gne enzyme (UDP-N-acetylgalactosamine 4-epimerase) necessary for the formation of UDP-galactosamine in these strains. By using rapid amplification of cDNA ends we were able to identify a transcription start site upstream of the terminal wzz gene, which showed differential transcription depending on the growth temperature of the strain. The Wzz protein is able to regulate the O34-antigen LPS chain length. The differential expression of this protein at different temperatures, which was substantially greater at 20 degrees C than at 37 degrees C, explains the previously observed differential production of O34-antigen LPS and its correlation with the virulence of A. hydrophila serotype O34 strains. PMID- 18408023 TI - Inactivation of lsr2 results in a hypermotile phenotype in Mycobacterium smegmatis. AB - Mycobacterial species are characterized by the presence of lipid-rich, hydrophobic cell envelopes. These cell envelopes contribute to properties such as roughness of colonies, aggregation of cells in liquid culture without detergent, and biofilm formation. We describe here a mutant strain of Mycobacterium smegmatis, called DL1215, which demonstrates marked deviations from the above mentioned phenotypes. DL1215 arose spontaneously from a strain deficient for the stringent response (M. smegmatis Delta rel(Msm) strain) and is not a reversion to a wild-type phenotype. The nature of the spontaneous mutation was a single base pair deletion in the lsr2 gene, leading to the formation of a truncated protein product. The DL1215 strain was complicated by having both inactivated rel(Msm) and lsr2 genes, and so a single lsr2 mutant was created to analyze the gene's function. The lsr2 gene was inactivated in the wild-type M. smegmatis mc(2)155 strain by allelic replacement to create strain DL2008. Strain DL2008 shows characteristics unique from those of both the wild-type and Delta rel(Msm) strains, some of which include a greatly enhanced ability to slide over agar surfaces (referred to here as "hypermotility"), greater resistance to phage infection and to the antibiotic kanamycin, and an inability to form biofilms. Complementation of the DL2008 mutant with a plasmid containing lsr2 (pLSR2) reverts the strain to the mc(2)155 phenotype. Although these phenotypic differences allude to changes in cell surface lipids, no difference is observed in glycopeptidolipids, polar lipids, apolar lipids, or mycolic acids of the cell wall. PMID- 18408024 TI - Protective action of ppGpp in microcin J25-sensitive strains. AB - As Escherichia coli strains enter the stationary phase of growth they become more resistant to the peptide antibiotic microcin J25. It is known that starvation for nutrients such as amino acids or glucose leads to increases in guanosine 3',5' bispyrophosphate (ppGpp) levels and that the intracellular concentration of this nucleotide increases as cells enter the stationary phase of growth. Therefore, we examined the effects of artificially manipulating the ppGpp levels on sensitivity to microcin J25. A direct correlation was found between ppGpp accumulation and microcin resistance. Our results indicate that the nucleotide is required to induce production of YojI, a chromosomally encoded efflux pump which, in turn, expels microcin from cells. This would maintain the intracellular level of the antibiotic below a toxic level. PMID- 18408025 TI - Genetic and biochemical characterization of the poly(3-hydroxybutyrate-co-3 hydroxyvalerate) synthase in Haloferax mediterranei. AB - The haloarchaeon Haloferax mediterranei has shown promise for the economical production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a desirable bioplastic. However, little is known at present about the genes involved in PHBV synthesis in the domain Archaea. In this study, we cloned the gene cluster (phaEC(Hme)) encoding a polyhydroxyalkanoate (PHA) synthase in H. mediterranei CGMCC 1.2087 via thermal asymmetric interlaced PCR. Western blotting revealed that the phaE(Hme) and phaC(Hme) genes were constitutively expressed, and both the PhaE(Hme) and PhaC(Hme) proteins were strongly bound to the PHBV granules. Interestingly, CGMCC 1.2087 could synthesize PHBV in either nutrient-limited medium (supplemented with 1% starch) or nutrient-rich medium, up to 24 or 18% (wt/wt) in shaking flasks. Knockout of the phaEC(Hme) genes in CGMCC 1.2087 led to a complete loss of PHBV synthesis, and only complementation with the phaEC(Hme) genes together (but not either one alone) could restore to this mutant the capability for PHBV accumulation. The known haloarchaeal PhaC subunits are much longer at their C termini than their bacterial counterparts, and the C terminal extension of PhaC(Hme) was proven to be indispensable for its function in vivo. Moreover, the mixture of purified PhaE(Hme)/PhaC(Hme) (1:1) showed significant activity of PHA synthase in vitro. Taken together, our results indicated that a novel member of the class III PHA synthases, composed of PhaC(Hme) and PhaE(Hme), accounted for the PHBV synthesis in H. mediterranei. PMID- 18408026 TI - Negative regulation of quorum-sensing systems in Pseudomonas aeruginosa by ATP dependent Lon protease. AB - Lon protease, a member of the ATP-dependent protease family, regulates numerous cellular systems by degrading specific substrates. Here, we demonstrate that Lon is involved in the regulation of quorum-sensing (QS) signaling systems in Pseudomonas aeruginosa, an opportunistic human pathogen. The organism has two acyl-homoserine lactone (HSL)-mediated QS systems, LasR/LasI and RhlR/RhlI. Many reports have demonstrated that these two systems are regulated and interconnected by global regulators. We found that lon-disrupted cells overproduce pyocyanin, the biosynthesis of which depends on the RhlR/RhlI system, and show increased levels of a transcriptional regulator, RhlR. The QS systems are organized hierarchically: the RhlR/RhlI system is subordinate to LasR/LasI. To elucidate the mechanism by which Lon negatively regulates RhlR/RhlI, we examined the effect of lon disruption on the LasR/LasI system. We found that Lon represses the expression of LasR/LasI by degrading LasI, an HSL synthase, leading to negative regulation of the RhlR/RhlI system. RhlR/RhlI was also shown to be regulated by Lon independently of LasR/LasI via regulation of RhlI, an HSL synthase. In view of these findings, it is suggested that Lon protease is a powerful negative regulator of both HSL-mediated QS systems in P. aeruginosa. PMID- 18408027 TI - Mutations in the scaffoldin gene, cipA, of Clostridium thermocellum with impaired cellulosome formation and cellulose hydrolysis: insertions of a new transposable element, IS1447, and implications for cellulase synergism on crystalline cellulose. AB - Mutants of Clostridium thermocellum that had lost the ability to adhere to microcrystalline cellulose were isolated. Six of them that showed diminished ability to depolymerize crystalline cellulose were selected. Size exclusion chromatography of the proteins from the culture supernatant revealed the loss of the supramolecular enzyme complex, the cellulosome. However, denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis resulted in extracellular protein patterns comparable to those of isolated cellulosomes, except for a missing CipA band. Sequencing of the six mutant cipA genes revealed a new insertion (IS) element, IS1447, belonging to the IS3 family. It was inserted into the cipA reading frame in four different locations: cohesin module 1, two different positions in the carbohydrate binding module, and cohesin module 3. The IS sequences were identical and consisted of a transposase gene and the inverted repeats IRR and IRS. The insertion resulted in an obviously nonspecific duplication of 3 base pairs within the target sequence. This lack of specificity allows transposition without the need of a defined target DNA sequence. Eighteen copies of IS1447 were identified in the genomic sequence of C. thermocellum ATCC 27405. At least one of them can be activated for transposition. Compared to the wild type, the mutant culture supernatant, with a completely defective CipA protein, showed equal specific hydrolytic activity against soluble beta-glucan but a 15-fold reduction in specific activity with crystalline cellulose. These results identify a genetic basis for the synergistic effect of complex formation on crystalline-cellulose degradation. PMID- 18408028 TI - The peptidoglycan of stationary-phase Mycobacterium tuberculosis predominantly contains cross-links generated by L,D-transpeptidation. AB - Our understanding of the mechanisms used by Mycobacterium tuberculosis to persist in a "dormant" state is essential to the development of therapies effective in sterilizing tissues. Gene expression profiling in model systems has revealed a complex adaptive response thought to endow M. tuberculosis with the capacity to survive several months of combinatorial antibiotic treatment. We show here that this adaptive response may involve remodeling of the peptidoglycan network by substitution of 4-->3 cross-links generated by the D,D-transpeptidase activity of penicillin-binding proteins by 3-->3 cross-links generated by a transpeptidase of L,D specificity. A candidate gene, previously shown to be upregulated upon nutrient starvation, was found to encode an L,D-transpeptidase active in the formation of 3-->3 cross-links. The enzyme, Ldt(Mt1), was inactivated by carbapenems, a class of beta-lactam antibiotics that are poorly hydrolyzed by the M. tuberculosis beta-lactamases. Ldt(Mt1) and carbapenems may therefore represent a target and a drug family relevant to the eradication of persistent M. tuberculosis. PMID- 18408029 TI - Single gene deletions of mrpA to mrpG and mrpE point mutations affect activity of the Mrp Na+/H+ antiporter of alkaliphilic Bacillus and formation of hetero oligomeric Mrp complexes. AB - Mrp antiporters catalyze secondary Na(+)(Li(+))/H(+) antiport and/or K(+)/H(+) antiport that is physiologically important in diverse bacteria. An additional capacity for anion flux has been observed for a few systems. Mrp is unique among antiporters in that it requires all six or seven hydrophobic gene products (MrpA to MrpG) of the mrp operon for full antiporter activity, but MrpE has been reported to be dispensable. Here, the membrane complexes formed by Mrp proteins were examined using a cloned mrp operon from alkaliphilic Bacillus pseudofirmus OF4. The operon was engineered so that the seven Mrp proteins could be detected in single samples. Membrane extracts of an antiporter-deficient Escherichia coli strain expressing this construct were analyzed by blue native-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Mrp complexes of two sizes were identified containing all seven Mrp proteins. Studies of the single nonpolar mrp gene deletions in the construct showed that a subcomplex of MrpA, MrpB, MrpC, and MrpD was formed in the absence of MrpE, MrpF, or MrpG. By contrast, MrpE, MrpF, and MrpG were not observed in membranes lacking MrpA, MrpB, MrpC, or MrpD. Although MrpA and MrpD have been hypothesized to be the antiporter proteins, the MrpA-to-D complex was inactive. Every Mrp protein was required for an activity level near that of the wild-type Na(+)/H(+) antiporter, but a very low activity level was observed in the absence of MrpE. The introduction of an MrpE(P114G) mutation into the full Mrp complex led to antiport activity with a greatly increased apparent K(m) value for Na(+). The results suggested that interactions among the proteins of heterooligomeric Mrp complexes strongly impact antiporter properties. PMID- 18408030 TI - Surface viscoelasticity of individual gram-negative bacterial cells measured using atomic force microscopy. AB - The cell envelope of gram-negative bacteria is responsible for many important biological functions: it plays a structural role, it accommodates the selective transfer of material across the cell wall, it undergoes changes made necessary by growth and division, and it transfers information about the environment into the cell. Thus, an accurate quantification of cell mechanical properties is required not only to understand physiological processes but also to help elucidate the relationship between cell surface structure and function. We have used a novel, atomic force microscopy (AFM)-based approach to probe the mechanical properties of single bacterial cells by applying a constant compressive force to the cell under fluid conditions while measuring the time-dependent displacement (creep) of the AFM tip due to the viscoelastic properties of the cell. For these experiments, we chose a representative gram-negative bacterium, Pseudomonas aeruginosa PAO1, and we used regular V-shaped AFM cantilevers with pyramid-shaped and colloidal tips. We find that the cell response is well described by a three element mechanical model which describes an effective cell spring constant, k(1), and an effective time constant, tau, for the creep deformation. Adding glutaraldehyde, an agent that increases the covalent bonding of the cell surface, produced a significant increase in k(1) together with a significant decrease in tau. This work represents a new attempt toward the understanding of the nanomechanical properties of single bacteria while they are under fluid conditions, which could be of practical value for elucidating, for instance, the biomechanical effects of drugs (such as antibiotics) on pathogens. PMID- 18408031 TI - Role of differential expression of streptococcal arginine deiminase in inhibition of fimA expression in Porphyromonas gingivalis. AB - Streptococcus cristatus ArcA inhibits production of a major adhesin, FimA, in Porphyromonas gingivalis, a primary periodontal pathogen. In this study, we demonstrate the differential expression of arcA in two streptococcal species. The expression level of arcA in streptococci appears to be controlled by both cis and trans elements. PMID- 18408032 TI - Characterization of abn2 (yxiA), encoding a Bacillus subtilis GH43 arabinanase, Abn2, and its role in arabino-polysaccharide degradation. AB - The extracellular depolymerization of arabinopolysaccharides by microorganisms is accomplished by arabinanases, xylanases, and galactanases. Here, we characterize a novel endo-alpha-1,5-l-arabinanase (EC 3.2.1.99) from Bacillus subtilis, encoded by the yxiA gene (herein renamed abn2) that contributes to arabinan degradation. Functional studies by mutational analysis showed that Abn2, together with previously characterized AbnA, is responsible for the majority of the extracellular arabinan activity in B. subtilis. Abn2 was overproduced in Escherichia coli, purified from the periplasmic fraction, and characterized with respect to substrate specificity and biochemical and physical properties. With linear-alpha-1,5-l-arabinan as the preferred substrate, the enzyme exhibited an apparent K(m) of 2.0 mg ml(-1) and V(max) of 0.25 mmol min(-1) mg(-1) at pH 7.0 and 50 degrees C. RNA studies revealed the monocistronic nature of abn2. Two potential transcriptional start sites were identified by primer extension analysis, and both a sigma(A)-dependent and a sigma(H)-dependent promoter were located. Transcriptional fusion studies revealed that the expression of abn2 is stimulated by arabinan and pectin and repressed by glucose; however, arabinose is not the natural inducer. Additionally, trans-acting factors and cis elements involved in transcription were investigated. Abn2 displayed a control mechanism at a level of gene expression different from that observed with AbnA. These distinct regulatory mechanisms exhibited by two members of extracellular glycoside hydrolase family 43 (GH43) suggest an adaptative strategy of B. subtilis for optimal degradation of arabinopolysaccharides. PMID- 18408033 TI - Unusual, virulence plasmid-dependent growth behavior of Yersinia enterocolitica in three-dimensional collagen gels. AB - As a first approach to establishing a three-dimensional culture infection model, we studied the growth behavior of the extracellular pathogen Yersinia enterocolitica in three-dimensional collagen gels (3D-CoG). Surprisingly, we observed that plasmidless Y. enterocolitica was motile in the 3D-CoG in contrast to its growth in traditional motility agar at 37 degrees C. Motility at 37 degrees C was abrogated in the presence of the virulence plasmid pYV or the exclusive expression of the pYV-located Yersinia adhesion gene yadA. YadA producing yersiniae formed densely packed (dp) microcolonies, whereas pYVDelta yadA-carrying yersiniae formed loosely packed microcolonies at 37 degrees C in 3D CoG. Furthermore, we demonstrated that the packing density of the microcolonies was dependent on the head domain of YadA. Moreover, dp microcolony formation did not depend on the capacity of YadA to bind to collagen fibers, as demonstrated by the use of yersiniae producing collagen nonbinding YadA. By using a yopE-gfp reporter, we demonstrated Ca(2+)-dependent expression of this pYV-localized virulence gene by yersiniae in 3D-CoG. In conclusion, this study revealed unique plasmid-dependent growth behavior of yersiniae in a three-dimensional matrix environment that resembles the behavior of yersiniae (e.g., formation of microcolonies) in infected mouse tissue. Thus, this 3D-CoG model may be a first step to a more complex level of in vitro infection models that mimic living tissue, enabling us to study the dynamics of pathogen-host cell interactions. PMID- 18408034 TI - Complete genome sequence of the soil actinomycete Kocuria rhizophila. AB - The soil actinomycete Kocuria rhizophila belongs to the suborder Micrococcineae, a divergent bacterial group for which only a limited amount of genomic information is currently available. K. rhizophila is also important in industrial applications; e.g., it is commonly used as a standard quality control strain for antimicrobial susceptibility testing. Sequencing and annotation of the genome of K. rhizophila DC2201 (NBRC 103217) revealed a single circular chromosome (2,697,540 bp; G+C content of 71.16%) containing 2,357 predicted protein-coding genes. Most of the predicted proteins (87.7%) were orthologous to actinobacterial proteins, and the genome showed fairly good conservation of synteny with taxonomically related actinobacterial genomes. On the other hand, the genome seems to encode much smaller numbers of proteins necessary for secondary metabolism (one each of nonribosomal peptide synthetase and type III polyketide synthase), transcriptional regulation, and lateral gene transfer, reflecting the small genome size. The presence of probable metabolic pathways for the transformation of phenolic compounds generated from the decomposition of plant materials, and the presence of a large number of genes associated with membrane transport, particularly amino acid transporters and drug efflux pumps, may contribute to the organism's utilization of root exudates, as well as the tolerance to various organic compounds. PMID- 18408035 TI - Investigating early events in receptor binding and translocation of colicin E9 using synchronized cell killing and proteolytic cleavage. AB - Enzymatic colicins such as colicin E9 (ColE9) bind to BtuB on the cell surface of Escherichia coli and rapidly recruit a second coreceptor, either OmpF or OmpC, through which the N-terminal natively disordered region (NDR) of their translocation domain gains entry into the cell periplasm and interacts with TolB. Previously, we constructed an inactive disulfide-locked mutant ColE9 (ColE9(s-s)) that binds to BtuB and can be reduced with dithiothreitol (DTT) to synchronize cell killing. By introducing unique enterokinase (EK) cleavage sites in ColE9(s s), we showed that the first 61 residues of the NDR were inaccessible to cleavage when bound to BtuB, whereas an EK cleavage site inserted at residue 82 of the NDR remained accessible. This suggests that most of the NDR is occluded by OmpF shortly after binding to BtuB, whereas the extreme distal region of the NDR is surface exposed before unfolding of the receptor-binding domain occurs. EK cleavage of unique cleavage sites located in the ordered region of the translocation domain or in the distal region of the receptor-binding domain confirmed that these regions of ColE9 remained accessible at the E. coli cell surface. Lack of EK cleavage of the DNase domain of the cell-bound, oxidized ColE9/Im9 complex, and the rapid detection of Alexa Fluor 594-labeled Im9 (Im9(AF)) in the cell supernatant following treatment of cells with DTT, suggested that immunity release occurred immediately after unfolding of the colicin and was not driven by binding to BtuB. PMID- 18408036 TI - Transcriptional regulation of the nitrile hydratase gene cluster in Pseudomonas chlororaphis B23. AB - An enormous amount of nitrile hydratase (NHase) is inducibly produced by Pseudomonas chlororaphis B23 after addition of methacrylamide as the sole nitrogen source to a medium. The expression pattern of the P. chlororaphis B23 NHase gene cluster in response to addition of methacrylamide to the medium was investigated. Recently, we reported that the NHase gene cluster comprises seven genes (oxdA, amiA, nhpA, nhpB, nhpC, nhpS, and acsA). Sequence analysis of the 1.5-kb region upstream of the oxdA gene revealed the presence of a 936-bp open reading frame (designated nhpR), which should encode a protein with a molecular mass of 35,098. The deduced amino acid sequence of the nhpR product showed similarity to the sequences of transcriptional regulators belonging to the XylS/AraC family. Although the transcription of the eight genes (nhpR, oxdA, amiA, nhpABC, nhpS, and acsA) in the NHase gene cluster was induced significantly in the P. chlororaphis B23 wild-type strain after addition of methacrylamide to the medium, transcription of these genes in the nhpR disruptant was not induced, demonstrating that nhpR codes for a positive transcriptional regulator in the NHase gene cluster. A reverse transcription-PCR experiment revealed that five genes (oxdA, amiA, nhpA, nhpB, and nhpC) are cotranscribed, as are two other genes (nhpS and acsA). The transcription start sites for nhpR, oxdA, nhpA, and nhpS were mapped by primer extension analysis, and putative -12 and -24 sigma(54) type promoter binding sites were identified. NhpR was found to be the first transcriptional regulator of NHase belonging to the XylS/AraC family. PMID- 18408037 TI - Pathway and endpoint free energy calculations for cyclic nucleotide binding to HCN channels. AB - cAMP and cGMP differentially bind to and regulate a variety of proteins, including cyclic nucleotide-gated (CNG) channels and hyperpolarization-activated cyclic nucleotide-regulated (HCN) channels. Previous site-directed mutagenesis studies have isolated two conserved residues that are critical for enabling certain channels to selectively bind cGMP relative to cAMP. However, no definitive mechanism has been identified that explains the preferential activation of other channels by cAMP. Here we apply computational binding free energy methods, including thermodynamic integration, linear interaction energy, and continuum electrostatic calculations, to gain insights into the mechanisms of cyclic nucleotide selectivity. Consistent with experimental observations, computational results for the cAMP-selective HCN channels show that the binding free energy of cAMP is lower (more favorable) than that of cGMP. Surprisingly, cAMP selectivity is not due to its preferential contacts with protein, but rather reflects the greater hydration energy of cGMP relative to cAMP, resulting in a greater energetic cost for cGMP binding. PMID- 18408038 TI - Dynamics of cellular focal adhesions on deformable substrates: consequences for cell force microscopy. AB - Cell focal adhesions are micrometer-sized aggregates of proteins that anchor the cell to the extracellular matrix. Within the cell, these adhesions are connected to the contractile, actin cytoskeleton; this allows the adhesions to transmit forces to the surrounding matrix and makes the adhesion assembly sensitive to the rigidity of their environment. In this article, we predict the dynamics of focal adhesions as a function of the rigidity of the substrate. We generalize previous theories and include the fact that the dynamics of proteins that adsorb to adhesions are also driven by their coupling to cell contractility and the deformation of the matrix. We predict that adhesions reach a finite size that is proportional to the elastic compliance of the substrate, on a timescale that also scales with the compliance: focal adhesions quickly reach a relatively small, steady-state size on soft materials. However, their apparent sliding is not sensitive to the rigidity of the substrate. We also suggest some experimental probes of these ideas and discuss the nature of information that can be extracted from cell force microscopy on deformable substrates. PMID- 18408039 TI - Regulation of the micromechanical properties of pulmonary endothelium by S1P and thrombin: role of cortactin. AB - Disruption of pulmonary endothelial cell (EC) barrier function is a critical pathophysiologic event in highly morbid inflammatory conditions such as sepsis and acute respiratory disease stress syndrome. Actin cytoskeleton, an essential regulator of endothelial permeability, is a dynamic structure whose stimuli induced rearrangement is linked to barrier modulation. Here, we used atomic force microscopy to characterize structural and mechanical changes in the F-actin cytoskeleton of cultured human pulmonary artery EC in response to both barrier enhancing (induced by sphingosine 1-phosphate (S1P)) and barrier-disrupting (induced by thrombin) conditions. Atomic force microscopy elasticity measurements show differential effects: for the barrier protecting molecule S1P, the elastic modulus was elevated significantly on the periphery; for the barrier-disrupting molecule thrombin, on the other hand, it was elevated significantly in the central region of the cell. The force and elasticity maps correlate with F-actin rearrangements as identified by immunofluorescence analysis. Significantly, reduced expression (via siRNA) of cortactin, an actin-binding protein essential to EC barrier regulation, resulted in a shift in the S1P-mediated elasticity pattern to more closely resemble control, unstimulated endothelium. PMID- 18408040 TI - The beta-strand-loop-beta-strand conformation is marginally populated in beta2 microglobulin (20-41) peptide in solution as revealed by replica exchange molecular dynamics simulations. AB - Solid-state NMR study shows that the 22-residue K3 peptide (Ser(20)-Lys(41)) from beta(2)-microglobulin (beta(2)m) adopts a beta-strand-loop-beta-strand conformation in its fibril state. Residue Pro(32) has a trans conformation in the fibril state of the peptide, while it adopts a cis conformation in the native state of full-length beta(2)m. To get insights into the structural properties of the K3 peptide, and determine whether the strand-loop-strand conformation is encoded at the monomeric level, we run all-atom explicit solvent replica exchange molecular dynamics on both the cis and trans variants. Our simulations show that the conformational space of the trans- and cis-K3 peptides is very different, with 1% of the sampled conformations in common at room temperature. In addition, both variants display only 0.3-0.5% of the conformations with beta-strand-loop beta-strand character. This finding, compared to results on the Alzheimer's Abeta peptide, suggests that the biases toward aggregation leading to the beta-strand loop-beta-strand conformation in fibrils are peptide-dependent. PMID- 18408041 TI - Molecular dynamics study of talin-vinculin binding. AB - Cells can sense mechanical force in regulating focal adhesion assembly. One vivid example is the force-induced recruitment of vinculin to reinforce initial contacts between a cell and the extracellular matrix. Crystal structures of the unbound proteins and bound complex between the vinculin head subdomain (Vh1) and the talin vinculin binding site 1 (VBS1) indicate that vinculin undergoes a conformational change upon binding to talin. However, the molecular basis for this event and the precise nature of the binding pathway remain elusive. In this article, molecular dynamics is used to investigate the binding mechanism of Vh1 and VBS1 under minimal constraints to facilitate binding. One simulation demonstrates binding of the two molecules in the complete absence of external force. VBS1 makes early hydrophobic contact with Vh1 by positioning the critical hydrophobic residues (L608, L615, and L622) in the groove formed by helices 1 and 2 of Vh1. The solvent-exposed hydrophobic residues (V619 and L623) then gradually penetrate the hydrophobic core of Vh1, thus further separating helix 1 from helix 2. These critical residues are highly conserved as large hydrophobic side groups in other vinculin binding sites; studies also have demonstrated that these residues are essential in Vh1-VBS1 binding. Similar binding mechanisms are also demonstrated in separate molecular dynamics simulations of Vh1 binding to other vinculin binding sites both in talin and alpha-actinin. PMID- 18408042 TI - Active mechanisms are needed to describe cell responses to submicrosecond, megavolt-per-meter pulses: cell models for ultrashort pulses. AB - Intracellular effects of submicrosecond, megavolt-per-meter pulses imply changes in a cell's plasma membrane (PM) and organelle membranes. The maximum reported PM transmembrane voltage is only 1.6 V and phosphatidylserine is translocated to the outer membrane leaflet of the PM. Passive membrane models involve only displacement currents and predict excessive PM voltages (approximately 25 V). Here we use a cell system model with nonconcentric circular PM and organelle membranes to demonstrate fundamental differences between active (nonlinear) and passive (linear) models. We assign active or passive interactions to local membrane regions. The resulting cell system model involves a large number of interconnected local models that individually represent the 1), passive conductive and dielectric properties of aqueous electrolytes and membranes; 2), resting potential source; and 3), asymptotic membrane electroporation model. Systems with passive interactions cannot account for key experimental observations. Our active models exhibit supra-electroporation of the PM and organelle membranes, some key features of the transmembrane voltage, high densities of small pores in the PM and organelle membranes, and a global postpulse perturbation in which cell membranes are depolarized on the timescale of pore lifetimes. PMID- 18408043 TI - Functional redundancy and new roles for genes of the autonomous floral-promotion pathway. AB - The early-flowering habit of rapid-cycling accessions of Arabidopsis (Arabidopsis thaliana) is, in part, due to the genes of the autonomous floral-promotion pathway (AP). The AP promotes flowering by repressing expression of the floral inhibitor FLOWERING LOCUS C (FLC). AP mutants are therefore late flowering due to elevated levels of FLC, and this late-flowering phenotype is eliminated by loss of-function mutations in FLC. To further investigate the role of the AP, we created a series of double mutants. In contrast to the phenotypes of single mutants, which are largely limited to delayed flowering, a subset of AP double mutants show a range of defects in growth and development. These phenotypes include reduced size, chlorophyll content, growth rate, and fertility. Unlike the effects of the AP on flowering time, these phenotypes are FLC independent. Recent work has also shown that two AP genes, FCA and FPA, are required for the repression and, in some cases, proper DNA methylation of two transposons. We show that similar effects are seen for all AP genes tested. Microarray analysis of gene expression in AP single and double mutants, however, suggests that the AP is not likely to play a broad role in the repression of gene expression through DNA methylation: very few of the genes that have been reported to be up-regulated in DNA methylation mutants are misexpressed in AP mutants. Together, these data indicate that the genes of the AP play important and sometimes functionally redundant roles in aspects of development in addition to flowering time. PMID- 18408044 TI - Naphthoquinone-dependent generation of superoxide radicals by quinone reductase isolated from the plasma membrane of soybean. AB - Using a tetrazolium-based assay, a NAD(P)H oxidoreductase was purified from plasma membranes prepared from soybean (Glycine max) hypocotyls. The enzyme, a tetramer of 85 kD, produces O2(.-) by a reaction that depended on menadione or several other 1,4-naphthoquinones, in apparent agreement with a classification as a one-electron-transferring flavoenzyme producing semiquinone radicals. However, the enzyme displayed catalytic and molecular properties of obligatory two electron-transferring quinone reductases of the DT-diaphorase type, including insensitivity to inhibition by diphenyleneiodonium. This apparent discrepancy was clarified by investigating the pH-dependent reactivity of menadionehydroquinone toward O2 and identifying the protein by mass spectrometry and immunological techniques. The enzyme turned out to be a classical NAD(P)H:quinone-acceptor oxidoreductase (EC 1.6.5.2, formerly 1.6.99.2) that reduces menadione to menadionehydroquinone and subsequently undergoes autoxidation at pH > or = 6.5. Autoxidation involves the production of the semiquinone as an intermediate, creating the conditions for one-electron reduction of O2. The possible function of this enzyme in the generation of O2(.-) and H2O2 at the plasma membrane of plants in vivo is discussed. PMID- 18408045 TI - Tobacco mosaic virus movement protein interacts with green fluorescent protein tagged microtubule end-binding protein 1. AB - The targeting of the movement protein (MP) of Tobacco mosaic virus to plasmodesmata involves the actin/endoplasmic reticulum network and does not require an intact microtubule cytoskeleton. Nevertheless, the ability of MP to facilitate the cell-to-cell spread of infection is tightly correlated with interactions of the protein with microtubules, indicating that the microtubule system is involved in the transport of viral RNA. While the MP acts like a microtubule-associated protein able to stabilize microtubules during late infection stages, the protein was also shown to cause the inactivation of the centrosome upon expression in mammalian cells, thus suggesting that MP may interact with factors involved in microtubule attachment, nucleation, or polymerization. To further investigate the interactions of MP with the microtubule system in planta, we expressed the MP in the presence of green fluorescent protein (GFP)-fused microtubule end-binding protein 1a (EB1a) of Arabidopsis (Arabidopsis thaliana; AtEB1a:GFP). The two proteins colocalize and interact in vivo as well as in vitro and exhibit mutual functional interference. These findings suggest that MP interacts with EB1 and that this interaction may play a role in the associations of MP with the microtubule system during infection. PMID- 18408046 TI - Roles of phosphatidylinositol 3-kinase in root hair growth. AB - The root hair is a model system for understanding plant cell tip growth. As phosphatidylinositol 3-phosphate [PtdIns(3)P] has been shown in other plant cell types to regulate factors that affect root hair growth, including reactive oxygen species (ROS) levels, cytoskeleton, and endosomal movement, we hypothesized that PtdIns(3)P is also important for root hair elongation. The enzyme that generates PtdIns(3)P, phosphatidylinositol 3-kinase (PI3K), was expressed in root hair cells of transgenic plants containing the PI3K promoter:beta-glucuronidase reporter construct. To obtain genetic evidence for the role of PtdIns(3)P in root hair elongation, we attempted to isolate Arabidopsis (Arabidopsis thaliana) mutant plants that did not express the gene VPS34 encoding the PI3K enzyme. However, the homozygous mutant was lethal due to gametophytic defects, and heterozygous plants were not discernibly different from wild-type plants. Alternatively, we made transgenic plants expressing the PtdIns(3)P-binding FYVE domain in the root hair cell to block signal transduction downstream of PtdIns(3)P. These transgenic plants had shorter root hairs and a reduced hair growth rate compared with wild-type plants. In addition, LY294002, a PI3K specific inhibitor, inhibited root hair elongation but not initiation. In LY294002-treated root hair cells, endocytosis at the stage of final fusion of the late endosomes to the tonoplast was inhibited and ROS level decreased in a dose dependent manner. Surprisingly, the LY294002 effects on ROS and root hair elongation were similar in rhd2 mutant plants, suggesting that RHD2 was not the major ROS generator in the PtdIns(3)P-mediated root hair elongation process. Collectively, these results suggest that PtdIns(3)P is required for maintenance of the processes essential for root hair cell elongation. PMID- 18408047 TI - Regulation of dormancy in barley by blue light and after-ripening: effects on abscisic acid and gibberellin metabolism. AB - White light strongly promotes dormancy in freshly harvested cereal grains, whereas dark and after-ripening have the opposite effect. We have analyzed the interaction of light and after-ripening on abscisic acid (ABA) and gibberellin (GA) metabolism genes and dormancy in barley (Hordeum vulgare 'Betzes'). Analysis of gene expression in imbibed barley grains shows that different ABA metabolism genes are targeted by white light and after-ripening. Of the genes examined, white light promotes the expression of an ABA biosynthetic gene, HvNCED1, in embryos. Consistent with this result, enzyme-linked immunosorbent assays show that dormant grains imbibed under white light have higher embryo ABA content than grains imbibed in the dark. After-ripening has no effect on expression of ABA biosynthesis genes, but promotes expression of an ABA catabolism gene (HvABA8'OH1), a GA biosynthetic gene (HvGA3ox2), and a GA catabolic gene (HvGA2ox3) following imbibition. Blue light mimics the effects of white light on germination, ABA levels, and expression of GA and ABA metabolism genes. Red and far-red light have no effect on germination, ABA levels, or HvNCED1. RNA interference experiments in transgenic barley plants support a role of HvABA8'OH1 in dormancy release. Reduced HvABA8'OH1 expression in transgenic HvABA8'OH1 RNAi grains results in higher levels of ABA and increased dormancy compared to nontransgenic grains. PMID- 18408048 TI - Cost-effectiveness of using continuous positive airway pressure in the treatment of severe obstructive sleep apnoea/hypopnoea syndrome in the UK. AB - OBJECTIVE: A study was undertaken to estimate the cost-effectiveness of using continuous positive airway pressure (CPAP) in the management of patients with severe obstructive sleep apnoea/hypopnoea syndrome (OSAHS) compared with no treatment from the perspective of the UK's National Health Service (NHS). METHODS: A Markov model was constructed to assess the cost-effectiveness of CPAP compared with no treatment. The model depicted the management of a 55-year-old patient with severe OSAHS as defined by an apnoea-hypopnoea index (AHI) >30 and daytime sleepiness (Epworth Sleepiness Scale score >or=12). The model spans a period of 14 years. RESULTS: According to the model, 57% of untreated patients are expected to be alive at the end of 14 years compared with 72% of patients treated with CPAP. Untreated patients are expected to cost the NHS pound10 645 (95% CI pound7988 to pound14,098) per patient over 14 years compared with pound9672 (95% CI pound8057 to pound12,860) per CPAP-treated patient. Treatment with CPAP for a period of 1 year was found not to be a cost-effective option since the cost per quality-adjusted life year (QALY) gained is expected to be > pound20,000, but after 2 years of treatment the cost per QALY gained is expected to be pound10,000 or less and, after 13 years of treatment, CPAP becomes a dominant treatment (ie, more effective than no treatment for less cost). CONCLUSION: Within the limitations of the model, CPAP was found to be clinically more effective than no treatment and, from the perspective of the UK's NHS, a cost-effective strategy after a minimum of 2 years of treatment. PMID- 18408049 TI - Randomised, placebo controlled trial of nebulised furosemide for breathlessness in patients with cancer. AB - BACKGROUND: Breathlessness is a common and difficult symptom to treat in patients with cancer. Case reports suggest that nebulised furosemide can relieve breathlessness in such patients but few data are available. METHOD: Patients with primary or secondary lung cancer and a Dyspnoea Exertion Scale score of >or=3 were recruited. Following familiarisation, patients received either nebulised furosemide 40 mg or nebulised 0.9% saline under double blind conditions or no treatment, in random order on 3 consecutive days. Patients undertook number reading and arm exercise tests to assess breathlessness and its impact, and were asked to report subjective benefit and any preference between nebulised treatments. RESULTS: 15 patients took part. There were no differences between furosemide, saline and no treatment in the outcomes of the number reading test (eg, mean number read per breath was 6.7, 6.4 and 6.7, respectively) or arm exercise test (eg, mean Borg score at maximum equivalent workload was 2.3, 2.5 and 2.7, respectively). No adverse effects were reported, although there was a small fall in forced expiratory volume in 1 s and forced vital capacity following saline. Six patients considered that their breathlessness improved with nebulised treatment, three preferring saline, one furosemide and two reporting they were of equal benefit. CONCLUSIONS: Our findings do not support a beneficial effect from nebulised furosemide in patients with cancer related breathlessness. Listed on the National Research Register (N0170118249) and the UK Clinical Research Network Portfolio Database (1428). PMID- 18408050 TI - Clinical outcome following lung transplantation in patients with cystic fibrosis colonised with Burkholderia cepacia complex: results from two French centres. AB - BACKGROUND: Infection with Burkholderia cepacia complex (BCC) is a life threatening complication of cystic fibrosis (CF), often seen as a contraindication for lung transplantation. METHODS: A long term retrospective study was conducted of all patients with CF undergoing lung transplants from January 1990 to October 2006 in two French centres allowing transplantation in patients colonised with BCC. RESULTS: 22 of the 247 lung transplant patients with CF were infected with BCC (B. cenocepacia genomovar III (n = 8), B. multivorans genomovar II (n = 11), B. vietnamiensis genomovar V (n = 2) and B. stabilis genomovar IV (n = 1)). BCC colonisation was not associated with any significant excess mortality (HR 1.5, 95% CI 0.7 to 3.2; p = 0.58). However, early mortality rates tended to be higher in the BCC group than in the non-BCC group (3 month survival: 85% vs 95%, respectively; log rank p = 0.05). Univariate analysis showed that the risk of death was significantly higher for the eight patients infected with B. cenocepacia than for the other 14 colonised patients (HR 3.2, 95% CI 1.1 to 5.9; p = 0.04). None of the other risk factors tested-primary graft failure, late extubation, septicaemia-had a significant effect. The 5 year cumulative incidence rate of bronchiolitis obliterans syndrome was not significantly higher in the BCC group than in the non-BCC group (38% vs 24%, respectively; p = 0.35). CONCLUSION: Our results suggest that BCC infection with a non-genomovar III organism may not be associated with excess mortality after lung transplantation in patients with CF and should not be seen as sufficient reason to exclude lung transplantation. However, colonisation with B. cenocepacia remains potentially detrimental. PMID- 18408051 TI - Lifecourse predictors of adult respiratory function: results from the Newcastle Thousand Families Study. AB - BACKGROUND: Impaired development in utero is suggested to increase the risk of poor respiratory health in adulthood, although a consensus has not been reached. A possible explanation for discrepancies between previous studies is inconsistent controlling for potential confounding factors, particularly childhood infections. Also, little is known regarding the relative importance of factors operating at different stages of the lifecourse. We have used detailed longitudinal data from the Newcastle Thousand Families cohort to assess the impact of birth weight, and various other factors acting throughout the lifecourse, on predicting forced expiratory volume in 1 s (FEV(1)). METHODS: Detailed information was collected prospectively during childhood, including birth weight, childhood infections and socioeconomic circumstances. At age 49-51 years, 412 study members attended for clinical examination and measurement of FEV(1). These data were analysed in relation to a range of factors from across the lifecourse using linear regression models. RESULTS: After adjustment for all other significant variables, increasing birth weight, standardised for sex and gestational age (p = 0.011), being breast fed for more than 4 weeks (p = 0.017), less frequent childhood lower respiratory tract infections (LRTI) (p = 0.015), non- smoking (p<0.001), lower body fat percentage (p = 0.010), male sex (p<0.001), no history of asthma (p = 0.013) and greater adult height (p<0.001) were all independently associated with higher adult FEV(1). CONCLUSION: Adult lung function is influenced by numerous factors during an individual's lifetime, acting both directly and indirectly throughout the lifecourse. As expected, sex, height and smoking were the most important predictors of FEV(1), but birth weight, breast feeding and childhood LRTIs also contributed significantly. PMID- 18408052 TI - A novel actin-related protein is associated with daughter cell formation in Toxoplasma gondii. AB - Cell division in Toxoplasma gondii occurs by an unusual budding mechanism termed endodyogeny, during which twin daughters are formed within the body of the mother cell. Cytokinesis begins with the coordinated assembly of the inner membrane complex (IMC), which surrounds the growing daughter cells. The IMC is compiled of both flattened membrane cisternae and subpellicular filaments composed of articulin-like proteins attached to underlying singlet microtubules. While proteins that comprise the elongating IMC have been described, little is known about its initial formation. Using Toxoplasma as a model system, we demonstrate that actin-like protein 1 (ALP1) is partially redistributed to the IMC at early stages in its formation. Immunoelectron microscopy localized ALP1 to a discrete region of the nuclear envelope, on transport vesicles, and on the nascent IMC of the daughter cells prior to the arrival of proteins such as IMC-1. The overexpression of ALP1 under the control of a strong constitutive promoter disrupted the formation of the daughter cell IMC, leading to delayed growth and defects in nuclear and apicoplast segregation. Collectively, these data suggest that ALP1 participates in the formation of daughter cell membranes during cell division in apicomplexan parasites. PMID- 18408053 TI - Rtr1 is the Saccharomyces cerevisiae homolog of a novel family of RNA polymerase II-binding proteins. AB - Cells must rapidly sense and respond to a wide variety of potentially cytotoxic external stressors to survive in a constantly changing environment. In a search for novel genes required for stress tolerance in Saccharomyces cerevisiae, we identified the uncharacterized open reading frame YER139C as a gene required for growth at 37 degrees C in the presence of the heat shock mimetic formamide. YER139C encodes the closest yeast homolog of the human RPAP2 protein, recently identified as a novel RNA polymerase II (RNAPII)-associated factor. Multiple lines of evidence support a role for this gene family in transcription, prompting us to rename YER139C RTR1 (regulator of transcription). The core RNAPII subunits RPB5, RPB7, and RPB9 were isolated as potent high-copy-number suppressors of the rtr1Delta temperature-sensitive growth phenotype, and deletion of the nonessential subunits RPB4 and RPB9 hypersensitized cells to RTR1 overexpression. Disruption of RTR1 resulted in mycophenolic acid sensitivity and synthetic genetic interactions with a number of genes involved in multiple phases of transcription. Consistently, rtr1Delta cells are defective in inducible transcription from the GAL1 promoter. Rtr1 constitutively shuttles between the cytoplasm and nucleus, where it physically associates with an active RNAPII transcriptional complex. Taken together, our data reveal a role for members of the RTR1/RPAP2 family as regulators of core RNAPII function. PMID- 18408054 TI - Contributions of the response regulators Ssk1p and Skn7p in the pseudohyphal development, stress adaptation, and drug sensitivity of the opportunistic yeast Candida lusitaniae. AB - We recently characterized the histidine kinase receptor genes of Candida lusitaniae. For the present study, we have further investigated the role of SSK1 and SKN7, encoding response regulators. The results of functional analysis of mutants indicated that Ssk1p is involved in osmotolerance and pseudohyphal development, whereas Skn7p appears crucial for oxidative stress adaptation. PMID- 18408055 TI - Transcriptional repression by the Pho4 transcription factor controls the timing of SNZ1 expression. AB - Nutrient-sensing kinases play important roles for the yeast Saccharomyces cerevisiae to adapt to new nutrient conditions when the nutrient status changes. Our previous global gene expression analysis revealed that the Pho85 kinase, one of the yeast nutrient-sensing kinases, is involved in the changes in gene expression profiles when yeast cells undergo a diauxic shift. We also found that the stationary phase-specific genes SNZ1 and SNO1, which share a common promoter, are not properly induced when Pho85 is absent. To examine the role of the kinase in SNZ1/SNO1 regulation, we analyzed their expression during the growth of various yeast mutants, including those affecting Pho85 function or lacking the Pho4 transcription factor, an in vivo substrate of Pho85, and tested Pho4 binding by chromatin immunoprecipitation. Pho4 exhibits temporal binding to the SNZ1/SNO1 promoter to down-regulate the promoter activity, and a Deltapho4 mutation advances the timing of SNZ1/SNO1 expression. SNZ2, another member of the SNZ/SNO family, is expressed at an earlier growth stage than SNZ1, and Pho4 does not affect this timing, although Pho85 is required for SNZ2 expression. Thus, Pho4 appears to regulate the different timing of the expression of the SNZ/SNO family members. Pho4 binding to the SNZ1/SNO1 promoter is accompanied by alterations in chromatin structure, and Rpd3 histone deacetylase is required for the proper timing of SNZ1/SNO1 expression, while Asf1 histone chaperone is indispensable for their expression. These results imply that Pho4 plays positive and negative roles in transcriptional regulation, with both cases involving structural changes in its target chromatin. PMID- 18408056 TI - MPL1, a novel phosphatase with leucine-rich repeats, is essential for proper ERK2 phosphorylation and cell motility. AB - The novel Dictyostelium phosphatase MPL1 contains six leucine-rich repeats at the amino-terminal end and a phosphatase domain at the carboxyl end. Similarly architectured phosphatases exist among other protozoa, such as Entamoeba histolytica, Leishmania major, and Trypanosoma cruzi. MPL1 was strongly induced after 5 h of development; ablation by homologous recombination led to defective streaming and aggregation during development. In addition, cyclic AMP (cAMP) pulsed mpl1(-) cells showed reduced random and directional motility. At the molecular level, mpl1(-) cells displayed higher prestimulus and persistent poststimulus ERK2 phosphorylation in response to cAMP stimulation. Consistent with their phenotype of persistent ERK2 phosphorylation, mpl1(-) cells also displayed an aberrant pattern of cAMP production, resembling that of the regA(-) cells. Reintroduction of a full-length MPL1 into mpl1(-) cells restored aggregation, ERK2 regulation, random and directional motility, and cAMP production similar to wild-type cells. We propose that MPL1 is a novel phosphatase essential for proper regulation of ERK2 phosphorylation and optimal motility during development. PMID- 18408057 TI - Identification of mating type genes in the bipolar basidiomycetous yeast Rhodosporidium toruloides: first insight into the MAT locus structure of the Sporidiobolales. AB - Rhodosporidium toruloides is a heterothallic, bipolar, red yeast that belongs to the Sporidiobolales, an order within a major lineage of basidiomycetes, the Pucciniomycotina. In contrast to other basidiomycetes, considerably less is known about the nature of the mating type (MAT) loci that control sexual reproduction in this lineage. Three genes (RHA1, RHA2, and RHA3) encoding precursors of the MAT A1 pheromone (rhodotorucine A) were previously identified and formed the basis for a genome walking approach that led to the identification of additional MAT genes in complementary mating strains of R. toruloides. Two mating type specific alleles encoding a p21-activated kinase (PAK; Ste20 homolog) were found between the RHA2 and RHA3 genes, and identification in MAT A2 strains of a gene encoding a presumptive pheromone precursor enabled prediction of the structure of rhodotorucine a. In addition, a putative pheromone receptor gene (STE3 homolog) was identified upstream of RHA1. Analyses of genomic data from two closely related species, Sporobolomyces roseus and Sporidiobolus salmonicolor, identified syntenic regions that contain homologs of all the above-mentioned genes. Notably, six novel pheromone precursor genes were uncovered, which encoded, similarly to the RHA genes, multiple tandem copies of the peptide moiety. This suggests that this structure, which is unique among fungal lipopeptide pheromones, seems to be prevalent in red yeasts. Species comparisons provided evidence for a large, multigenic MAT locus structure in the Sporidiobolales, but no putative homeodomain transcription factor genes (which are present in all basidiomycetous MAT loci characterized thus far) could be found in any of the three species in the vicinity of the MAT genes identified. PMID- 18408058 TI - The nascent parasitophorous vacuole membrane of Encephalitozoon cuniculi is formed by host cell lipids and contains pores which allow nutrient uptake. AB - Microsporidia are obligate intracellular pathogens which enter host cells by the discharge of a hollow tube through which the sporoplasma is extruded into the host cell. Since this invasion mechanism is very different from common entry strategies, the formation of the parasitophorous vacuole (PV) in Encephalitozoon species is likely to be distinct from known principles. We investigated the origin of the nascent Encephalitozoon cuniculi PV membrane with the aid of fluorescent lipid probes. When Bodipy 500/510-C(12)-HPC-labeled spores were used for infection, the emerging PV membrane was unlabeled, suggesting that sporoplasma-derived lipids do not significantly contribute to the formation of the PV membrane. In contrast, when raft and nonraft microdomains of the host cell plasma membrane were selectively labeled with DiIC(16) and Speedy DiO, both tracers were detectable in the nascent PV membrane shortly after infection, indicating that the bulk lipids of the PV membrane are host cell derived. Time lapse fluorescence microscopy revealed that the formation of the PV membrane is a fast event (<1.3 s), which occurred simultaneously with the extrusion of the sporoplasma. The portion of the discharged tube which is in contact with the host cell was found to be coated with labeled host cell lipids, which might be an indication for a plasma membrane invagination at the contact site. To investigate the presence of pores in the E. cuniculi PV membrane, we microinjected fluorescent dyes of different sizes into infected host cells. A 0.5-kDa dextran as well as 0.8- to 1.1-kDa peptides could rapidly enter the PV, while a 10-kDa dextran was stably excluded from the PV lumen, indicating that the PV membrane possesses pores with an exclusion size of <10 kDa, which should allow metabolite exchange. PMID- 18408059 TI - Detection of active butyrate-degrading microorganisms in methanogenic sludges by RNA-based stable isotope probing. AB - Butyrate-degrading bacteria in four methanogenic sludges were studied by RNA based stable isotope probing. Bacterial populations in the (13)C-labeled rRNA fractions were distinct from unlabeled fractions, and Syntrophaceae species, Tepidanaerobacter sp., and Clostridium spp. dominated. These results suggest that diverse microbes were active in butyrate degradation under methanogenic conditions. PMID- 18408060 TI - The probiotic Lactobacillus johnsonii NCC 533 produces high-molecular-mass inulin from sucrose by using an inulosucrase enzyme. AB - Fructansucrase enzymes polymerize the fructose moiety of sucrose into levan or inulin fructans, with beta(2-6) and beta(2-1) linkages, respectively. The probiotic bacterium Lactobacillus johnsonii strain NCC 533 possesses a single fructansucrase gene (open reading frame AAS08734) annotated as a putative levansucrase precursor. However, (13)C nuclear magnetic resonance (NMR) analysis of the fructan product synthesized in situ revealed that this is of the inulin type. The ftf gene of L. johnsonii was cloned and expressed to elucidate its exact identity. The purified L. johnsonii protein was characterized as an inulosucrase enzyme, producing inulin from sucrose, as identified by (13)C NMR analysis. Thin-layer chromatographic analysis of the reaction products showed that InuJ synthesized, besides the inulin polymer, a broad range of fructose oligosaccharides. Maximum InuJ enzyme activity was observed in a pH range of 4.5 to 7.0, decreasing sharply at pH 7.5. InuJ exhibited the highest enzyme activity at 55 degrees C, with a drastic decrease at 60 degrees C. Calcium ions were found to have an important effect on enzyme activity and stability. Kinetic analysis showed that the transfructosylation reaction of the InuJ enzyme does not obey Michaelis-Menten kinetics. The non-Michaelian behavior of InuJ may be attributed to the oligosaccharides that were initially formed in the reaction and which may act as better acceptors than the growing polymer chain. This is only the second example of the isolation and characterization of an inulosucrase enzyme and its inulin (oligosaccharide) product from a Lactobacillus strain. Furthermore, this is the first Lactobacillus strain shown to produce inulin polymer in situ. PMID- 18408061 TI - The supernumerary chromosome of Nectria haematococca that carries pea pathogenicity-related genes also carries a trait for pea rhizosphere competitiveness. AB - Fungi are found in a wide range of environments, and the ecological and host diversity of the fungus Nectria haematococca has been shown to be due in part to unique genes on different supernumerary chromosomes. These chromosomes have been called "conditionally dispensable" (CD) since they are not needed for axenic growth but are important for expanding the host range of individual isolates. From a biological perspective, the CD chromosomes can be compared to bacterial plasmids that carry unique genes that can define the habits of these microorganisms. The current study establishes that the N. haematococca PDA1-CD chromosome, which contains the genes for pea pathogenicity (PEP cluster) on pea roots, also carries a gene(s) for the utilization of homoserine, a compound found in large amounts in pea root exudates. Competition studies demonstrate that an isolate that lacks the PEP cluster but carries a portion of the CD chromosome which includes the homoserine utilization (HUT) gene(s) is more competitive in the pea rhizosphere than an isolate without the CD chromosome. PMID- 18408062 TI - Free-living tube worm endosymbionts found at deep-sea vents. AB - Recent evidence suggests that deep-sea vestimentiferan tube worms acquire their endosymbiotic bacteria from the environment each generation; thus, free-living symbionts should exist. Here, free-living tube worm symbiont phylotypes were detected in vent seawater and in biofilms at multiple deep-sea vent habitats by PCR amplification, DNA sequence analysis, and fluorescence in situ hybridization. These findings support environmental transmission as a means of symbiont acquisition for deep-sea tube worms. PMID- 18408063 TI - In vitro mutagenesis of Bacillus subtilis by using a modified Tn7 transposon with an outward-facing inducible promoter. AB - A Tn7 donor plasmid, pTn7SX, was constructed for use with the model gram-positive bacterium Bacillus subtilis. This new mini-Tn7, mTn7SX, contains a spectinomycin resistance cassette and an outward-facing, xylose-inducible promoter, thereby allowing for the regulated expression of genes downstream of the transposon. We demonstrate that mTn7SX inserts are obtained at a high frequency and occur randomly throughout the B. subtilis genome. The utility of this system was demonstrated by the selection of mutants with increased resistance to the antibiotic fosfomycin or duramycin. PMID- 18408064 TI - Synthesis and accumulation of cyanophycin in transgenic strains of Saccharomyces cerevisiae. AB - Cyanophycin [multi-L-arginyl-poly(L-aspartic acid) (CGP)] was, for the first time, produced in yeast. As yeasts are very important production organisms in biotechnology, it was determined if CGP can be produced in two different strains of Saccharomyces cerevisiae. The episomal vector systems pESC (with the galactose inducible promoter GAL1) and pYEX-BX (with the copper ion-inducible promoter CUP1) were chosen to express the cyanophycin synthetase gene from the cyanobacterium Synechocystis sp. strain PCC 6308 (cphA(6308)) in yeast. Expression experiments with transgenic yeasts revealed that the use of the CUP1 promoter is much more efficient for CGP production than the GAL1 promoter. As observed by electrophoresis of isolated CGP in sodium dodecyl sulfate polyacrylamide gels, the yeast strains produced two different types of polymer: the water-soluble and the water-insoluble CGP were observed as major and minor forms of the polymer, respectively. A maximum CGP content of 6.9% (wt/wt) was detected in the cells. High-performance liquid chromatography analysis showed that the isolated polymers consisted mainly of the two amino acids aspartic acid and arginine and that, in addition, a minor amount (2 mol%) of lysine was present. Growth of transgenic yeasts in the presence of 15 mM lysine resulted in an incorporation of up to 10 mol% of lysine into CGP. Anti-CGP antibodies generated against CGP isolated from Escherichia coli TOP10 harboring cphA(6308) reacted with insoluble CGP but not with soluble CGP, if applied in Western or dot blots. PMID- 18408065 TI - Investigating the properties of Bacillus thuringiensis Cry proteins with novel loop replacements created using combinatorial molecular biology. AB - Cry proteins are a large family of crystalline toxins produced by Bacillus thuringiensis. Individually, the family members are highly specific, but collectively, they target a diverse range of insects and nematodes. Domain II of the toxins is important for target specificity, and three loops at its apex have been studied extensively. There is considerable interest in determining whether modifications in this region may lead to toxins with novel specificity or potency. In this work, we studied the effect of loop substitution on toxin stability and specificity. For this purpose, sequences derived from antibody complementarity-determining regions (CDR) were used to replace native domain II apical loops to create "Crybodies." Each apical loop was substituted either individually or in combination with a library of third heavy-chain CDR (CDR-H3) sequences to create seven distinct Crybody types. An analysis of variants from each library indicated that the Cry1Aa framework can tolerate considerable sequence diversity at all loop positions but that some sequence combinations negatively affect structural stability and protease sensitivity. CDR-H3 substitution showed that loop position was an important determinant of insect toxicity: loop 2 was essential for activity, whereas the effects of substitutions at loop 1 and loop 3 were sequence dependent. Unexpectedly, differences in toxicity did not correlate with binding to cadherins--a major class of toxin receptors--since all Crybodies retained binding specificity. Collectively, these results serve to better define the role of the domain II apical loops as determinants of specificity and establish guidelines for their modification. PMID- 18408066 TI - Development and use of an efficient system for random mariner transposon mutagenesis to identify novel genetic determinants of biofilm formation in the core Enterococcus faecalis genome. AB - Enterococcus faecalis is a gram-positive commensal bacterium of the gastrointestinal tract and an important opportunistic pathogen. Despite the increasing clinical significance of the enterococci, most of the genetic analysis of these organisms has focused on mobile genetic elements, and existing tools for manipulation and analysis of the core E. faecalis chromosome are limited. We are interested in a comprehensive analysis of the genetic determinants for biofilm formation encoded within the core E. faecalis genome. To identify such determinants, we developed a substantially improved system for transposon mutagenesis in E. faecalis based on a mini-mariner transposable element. Mutagenesis of wild-type E. faecalis with this element yielded predominantly mutants carrying a single copy of the transposable element, and insertions were distributed around the entire chromosome in an apparently random fashion. We constructed a library of E. faecalis transposon insertion mutants and screened this library to identify mutants exhibiting a defect in biofilm formation. Biofilm-defective mutants were found to carry transposon insertions both in genes that were previously known to play a role in biofilm formation and in new genes lacking any known function; for several genes identified in the screen, complementation analysis confirmed a direct role in biofilm formation. These results provide significant new information about the genetics of enterococcal biofilm formation and demonstrate the general utility of our transposon system for functional genomic analysis of E. faecalis. PMID- 18408067 TI - Isolation of fungal cellobiohydrolase I genes from sporocarps and forest soils by PCR. AB - Cellulose is the major component of plant biomass, and microbial cellulose utilization is a key step in the decomposition of plant detritus. Despite this, little is known about the diversity of cellulolytic microbial communities in soil. Fungi are well known for their cellulolytic activity and mediate key functions during the decomposition of plant detritus in terrestrial ecosystems. We developed new oligonucleotide primers for fungal exocellulase genes (cellobiohydrolase, cbhI) and used these to isolate distinct cbhI homologues from four species of litter-decomposing basidiomycete fungi (Clitocybe nuda, Clitocybe gibba, Clitopilus prunulus, and Chlorophyllum molybdites) and two species of ascomycete fungi (Xylaria polymorpha and Sarcoscypha occidentalis). Evidence for cbhI gene families was found in three of the four basidiomycete species. Additionally, we isolated and cloned cbhI genes from the forest floor and mineral soil of two upland forests in northern lower Michigan, one dominated by oak (Quercus velutina, Q. alba) and the other dominated by sugar maple (Acer saccharum) and American basswood (Tilia americana). Phylogenetic analysis demonstrated that cellobiohydrolase genes recovered from the floor of both forests tended to cluster with Xylaria or in one of two unidentified groups, whereas cellobiohydrolase genes recovered from soil tended to cluster with Trichoderma, Alternaria, Eurotiales, and basidiomycete sequences. The ability to amplify a key fungal gene involved in plant litter decomposition has the potential to unlock the identity and dynamics of the cellulolytic fungal community in situ. PMID- 18408068 TI - Purification and characterization of a new family 45 endoglucanase, STCE1, from Staphylotrichum coccosporum and its overproduction in Humicola insolens. AB - In the detergent industry, fungal endoglucanases have been used to release microfibrils (defibrillation) from the surface of dyed cellulosic fabrics to enhance color brightness. Although endoglucanases for laundry use must have various properties, such as a neutral or alkaline optimum pH, resistance to anionic surfactants and oxidizing agents (main components in detergents), and high defibrillation activity, all-purpose endoglucanases have not been obtained yet. As a result of screening of endoglucanases, a new family 45 endoglucanase (family 45 glycoside hydrolase), designated STCE1, was obtained and purified to apparent homogeneity from the culture supernatant of Staphylotrichum coccosporum NBRC 31817. The molecular mass of STCE1 was 49 kDa. The optimum pH for the carboxymethyl cellulase activity of STCE1 was 6.0, and the optimum temperature was 60 degrees C. STCE1 was highly resistant to an anionic surfactant and an oxidizing agent. Furthermore, the defibrillation activities on dyed cotton and lyocell fabrics of STCE1 were higher than those of the other representative endoglucanases tested. These results indicate that STCE1 is an all-purpose enzyme for laundry use. A gene encoding STCE1, designated the stce1 gene, was cloned from S. coccosporum, and the complete sequence was determined. STCE1 consisted of three distinct domains: an N-terminal catalytic domain (family 45), a linker domain, and a C-terminal carbohydrate-binding module (family 1). The amino acid sequences of the catalytic domain of STCE1 were phylogenetically close to those of the family 45 endoglucanases EGL3, EGL4, and EGV from a Humicola sp. Hence, the stce1 gene was transferred into Humicola insolens and expressed. As a result, extremely high levels (0.90 mg protein per ml of culture supernatant, 27% of the total proteins) of the recombinant STCE1 were secreted as a mature form in the culture supernatant. PMID- 18408069 TI - HD-ZIP III activity is modulated by competitive inhibitors via a feedback loop in Arabidopsis shoot apical meristem development. AB - Shoot apical meristem (SAM) development is coordinately regulated by two interdependent signaling events: one maintaining stem cell identity and the other governing the initiation of lateral organs from the flanks of the SAM. The signaling networks involved in this process are interconnected and are regulated by multiple molecular mechanisms. Class III homeodomain-leucine zipper (HD-ZIP III) proteins are the most extensively studied transcription factors involved in this regulation. However, how different signals are integrated to maintain stem cell identity and to pattern lateral organ polarity remains unclear. Here, we demonstrated that a small ZIP protein, ZPR3, and its functionally redundant homolog, ZPR4, negatively regulate the HD-ZIP III activity in SAM development. ZPR3 directly interacts with PHABULOSA (PHB) and other HD-ZIP III proteins via the ZIP motifs and forms nonfunctional heterodimers. Accordingly, a double mutant, zpr3-2 zpr4-2, exhibits an altered SAM activity with abnormal stem cell maintenance. However, the mutant displays normal patterning of leaf polarity. In addition, we show that PHB positively regulates ZPR3 expression. We therefore propose that HD-ZIP III activity in regulating SAM development is modulated by, among other things, a feedback loop involving the competitive inhibitors ZPR3 and ZPR4. PMID- 18408070 TI - Transgenic expression of matrix metalloproteinase-9 causes adult-onset emphysema in mice associated with the loss of alveolar elastin. AB - Matrix metalloproteinase (MMP)-9 has been consistently identified in the lungs of patients with chronic obstructive pulmonary disease (COPD). However, its role in the development of the disease remains undefined. Mice that specifically express human MMP-9 in their macrophages were generated, and morphometric, biochemical, and histological analyses were conducted on the transgenic and littermate control mice over 1 yr to determine the effect of macrophage MMP-9 expression on emphysema formation and lung matrix content. Lung morphometry was normal in transgenic mice at 2 mo of age (mean linear intercept = 50+/-3 littermate mice vs. 51+/-2 transgenic mice). However, after 12 mo of age, the MMP-9 transgenic mice developed significant air space enlargement (mean linear intercept = 53+/-3 littermate mice vs. 61+/-2 MMP-9 transgenic mice; P<0.04). Lung hydroxyproline content was not significantly different between wild-type and transgenic mice, but MMP-9 did significantly decrease alveolar wall elastin at 1 yr of age (4.9+/ 0.3% area of alveolar wall in the littermate mice vs. 3.3+/-0.3% area of alveolar wall in the MMP-9 mice; P<0.004). Thus these results establish a central role for MMP-9 in the pathogenesis of this disease by demonstrating that expression of this protease in macrophages can alter the extracellular matrix and induce progressive air space enlargement in mice. PMID- 18408072 TI - Five-year outcomes of severe acute kidney injury requiring renal replacement therapy. AB - BACKGROUND: Current research priorities in critical care medicine are focusing on long-term outcomes of survivors of critical illness. Severe acute kidney injury (AKI) is a common occurrence in intensive care. However, few studies have followed up these patients beyond 12 months after hospital discharge. METHODS: Of a cohort of 425 patients, 226 survivors with severe AKI necessitating renal replacement therapy (RRT) were followed up for 60 months after hospital discharge. None of these patients had pre-existing kidney disease. Vital status and renal function were documented annually for 5 years. RESULTS: None of the discharged or transferred patients was dependent on RRT; 57% had complete recovery and 43% had partial recovery of renal function. During the first year after hospital discharge, 18% of survivors died, during the second year 4% and during the third to fifth year 2% per year. At 5 years, 25% of the cohort were still alive. Further improvement in renal function (eGFR) was noted in 26 patients within the first year only. Deterioration of renal function occurred in eight patients. At 5 years, renal function was normal in 86% of the remaining survivors, it was impaired in 9% and 5% of the patients alive needed dialysis again. The proportional Cox regression analysis model showed that pre-existing extrarenal comorbidity, surgery and partial recovery of renal function were independent determinants of long-term survival. CONCLUSIONS: This prospective observational study indicates that severe AKI is not only a determinant of excess in-hospital case fatalities of critically ill patients, but it also carries significant implications for long-term mortality. PMID- 18408073 TI - A kidney from hell? A nephrological view of the Whitechapel murders in 1888. AB - In the poor Whitechapel district of the East End of London in the fall of 1888, at least five prostitutes were brutally murdered, and in all but one case, also mutilated. The murderer was never caught and became known by his nickname 'Jack the Ripper'. The left kidney and the uterus were cut out and taken away from one of the victims named Catherine Eddowes. A kidney was also cut out of the body from another victim, but not taken away. Two weeks later, George Lusk, president of the Whitechapel Vigilance Committee, received a small cardboard box with half of a longitudinally divided kidney and a letter entitled 'From hell' claiming that the kidney inside the box was taken from the victim. The kidney was brought to Dr Thomas Horrocks Openshaw, the Curator of the London Pathological Museum, where the kidney could be microscopically examined. The press jumped on the topic and made a circumstantial case that this kidney had been indeed torn from the body of Catherine Eddowes. According to the later memoirs of Major Henry Smith of the City Police published more than 20 years after the incident, the kidney left in the corpse of Catherine Eddowes was in an advanced stage of Bright's disease and the kidney sent to George Lusk was in exactly a similar stage. Today, the majority of criminologists believe that the kidney sent to Mr Lusk was a hoax as were other letters signed with Jack the Ripper. However, the murderer took organs from his victims, and in the case of Catherine Eddowes, the kidney. Serial killers often mutilate their victims and abscond with the removed body parts as trophies. By removing the kidney from Catherine Eddowes, Jack the Ripper may have tried to take possession of the conscience, emotions and desires of one of his victims, attributes residing in the kidney as described in the Bible. Jack the Ripper was never caught; many suspects have been suggested, and the murder series ended as suddenly as it had begun. We will never know who this mentally disturbed 'nephrophilic' was. Today, the story of Jack the Ripper is part of contemporary culture. PMID- 18408071 TI - GABAA receptors are expressed and facilitate relaxation in airway smooth muscle. AB - Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian central nervous system and exerts its actions via both ionotropic (GABA(A)) channels and metabotropic (GABA(B)) receptors. GABA(A) channels are ubiquitously expressed in neuronal tissues, and in mature neurons modulate an inward chloride current resulting in neuronal inhibition due to membrane hyperpolarization. In airway smooth muscle (ASM) cells, membrane hyperpolarization favors smooth muscle relaxation. Although GABA(A) channels and GABA(B) receptors have been functionally identified on peripheral nerves in the lung, GABA(A) channels have never been identified on ASM itself. We detected the mRNA encoding of the GABA(A) alpha(4)-, alpha(5)-, beta(3)-, delta-, gamma(1-3)-, pi-, and theta-subunits in total RNA isolated from native human and guinea pig ASM and from cultured human ASM cells. Selected immunoblots identified the GABA(A) alpha(4)-, alpha(5)-, beta(3)-, and gamma(2)-subunit proteins in native human and guinea pig ASM and cultured human ASM cells. The GABA(A) beta(3) subunit protein was immunohistochemically localized to ASM in guinea pig tracheal rings. While muscimol, a specific GABA(A) channel agonist, did not affect the magnitude or the time to peak contractile effect of substance P, it directly concentration dependently relaxed a tachykinin-induced contraction in guinea pig tracheal rings, which was inhibited by the GABA(A)-selective antagonist gabazine. Muscimol also relaxed a contraction induced by an alternative contractile agonist histamine. These results demonstrate that functional GABA(A) channels are expressed on ASM and suggest a novel therapeutic target for the relaxation of ASM in diseases such as asthma and chronic obstructive lung disease. PMID- 18408074 TI - Association between cytokine gene polymorphisms and outcomes in renal transplantation: a meta-analysis of individual patient data. AB - BACKGROUND: Cytokine gene polymorphisms have been associated with poor outcomes after renal transplantation such as chronic allograft nephropathy (CAN), graft rejection (GR) and graft failure (GF), but the effects of these polymorphisms are still controversial. We therefore conducted a systematic review, with individual patient data (IPD) where possible, to determine the association between cytokine polymorphisms (TGF-beta1, TNF-alpha and IL-10) and outcomes after renal transplantation. METHODS: Five investigators were willing to participate and provided IPD. The outcomes of interest were GF, GR and CAN. Subjects with at least one of these were classified as having poor outcomes. Heterogeneity of gene effects was assessed. Multiple logistic regression was applied to assess gene effects, adjusting for clinical variables such as HLA matching and age. RESULTS: One-thousand and eighty-seven subjects were included in the IPD meta-analysis. Pooled results showed no evidence of heterogeneity and indicated that the strongest variables determining poor outcomes are HLA mismatching (OR = 1.6-1.8 for >/=3 HLA-A, -B, -DR mismatches compared with those with <3 mismatches) and age (OR = 1.2-1.4 for age 45 years or more). Incremental information on risk of a poor outcome is provided by the TGF-beta1c10 polymorphism (OR = 1.5, P = 0.034, 95% CI: 1.0-2.2 for TC genotype compared to TT genotype). Haplotypes of TGF-beta1 at c10 and c25 were inferred and the C-C haplotype was a marker of a poor outcome (OR = 1.3, P = 0.177, 95% CI: 1.0-2.3). Three polymorphisms of the IL-10 gene at 1082, -819, -592 are in strong linkage disequilibrium with each other (correlation coefficients: 0.6-1) and inferred haplotypes between these three loci show some association, with ACC increasing the risk of poor events compared to GCC (OR = 1.3, P = 0.044, 95% CI: 0.9-1.6). CONCLUSION: Pooled results to date suggest possible association between both the TGF-beta1 c10 polymorphism and a 3 SNP-haplotype of IL-10 and poor outcomes in renal transplantation, but this needs to be confirmed in larger studies. PMID- 18408076 TI - Cinacalcet does not affect longitudinal growth but increases body weight gain in experimental uraemia. AB - BACKGROUND: Cinacalcet (CIN) efficiently suppresses parathyroid hormone (PTH) secretion by the activation of the calcium-sensing receptor (CaR). Epiphyseal chondrocytes also express the CaR and its activation promotes cell proliferation and differentiation in vitro. Hence, the impact of CIN on the growth plate function requires assessment before routine administration in children. METHODS: We treated subtotally nephrectomized (SNX) and sham-operated, ad lib and pair-fed Sprague-Dawley rats with CIN (15 mg/kg day) or solvent (S) for 14 days p.o. and assessed whole body and tibia length gain, growth plate morphology, osseous front advance (OFA) (calcein staining) and chondrocyte proliferation rate [5-bromo-2' deoxyuridine (BrdU) staining]. RESULTS: Total body length gain did not differ after 7 and 14 days (SNX + CIN 2.9 +/- 0.6, SNX + S 3.0 +/- 0.7; sham + CIN 4.2 +/- 0.4, sham + S 4.5 +/- 0.4; sham pair-fed + CIN 3.3 +/- 0.5, sham pair-fed + S 3.5 +/- 0.6 cm/14 days; P = n.s.). Tibia length, the height of the total growth plate and the hypertrophic zone, OFA and chondrocyte proliferation rate were similar with CIN and S. Serum Ca(2+) declined with CIN treatment; PTH was 61% lower in CIN- compared to S-treated SNX (P < 0.05). Food intake was similar, whereas body weight gain (21.6 +/- 8.7 versus 12.7 +/- 11.2 g) and body weight gain per food intake (141 +/- 50 versus 77 +/- 70 g/kg) improved in CIN- versus S treated SNX animals (P < 0.05). CONCLUSION: CIN treatment does not impact on growth plate chondrocyte function in uraemic rats, but improves food efficiency and body weight gain. PMID- 18408075 TI - Marked variation in the definition and diagnosis of delayed graft function: a systematic review. AB - BACKGROUND: The term delayed graft function (DGF) is commonly used to describe the need for dialysis after receiving a kidney transplant. DGF increases morbidity after transplantation, prolongs hospitalization and may lead to premature graft failure. Various definitions of DGF are used in the literature without a uniformly accepted technique to identify DGF. METHODS: We performed a systematic review of the literature to identify all of the different definitions and diagnostic techniques to identify DGF. RESULTS: We identified 18 unique definitions for DGF and 10 diagnostic techniques to identify DGF. CONCLUSIONS: The utilization of heterogeneous clinical criteria to define DGF has certain limitations. It will lead to delayed and sometimes inaccurate diagnosis of DGF. Hence a diagnostic test that identifies DGF reliably and early is necessary. Heterogeneity, in the definitions used for DGF, hinders the evolution of a diagnostic technique to identify DGF, which requires a gold standard definition. We are in need of a new definition that is uniformly accepted across the kidney transplant community. The new definition will be helpful in promoting better communication among transplant professionals and aids in comparing clinical studies of diagnostic techniques to identify DGF and thus may facilitate clinical trials of interventions for the treatment of DGF. PMID- 18408077 TI - Use of a renal-specific oral supplement by haemodialysis patients with low protein intake does not increase the need for phosphate binders and may prevent a decline in nutritional status and quality of life. AB - BACKGROUND: Protein-energy wasting is a frequent and debilitating condition in maintenance dialysis. We randomly tested if an energy-dense, phosphate restricted, renal-specific oral supplement could maintain adequate nutritional intake and prevent malnutrition in maintenance haemodialysis patients with insufficient intake. METHODS: Eighty-six patients were assigned to a standard care (CTRL) group or were prescribed two 125-ml packs of Renilon 7.5(R) daily for 3 months (SUPP). Dietary intake, serum (S) albumin, prealbumin, protein nitrogen appearance (nPNA), C-reactive protein, subjective global assessment (SGA) and quality of life (QOL) were recorded at baseline and after 3 months. RESULTS: While intention to treat analysis (ITT) did not reveal strong statistically significant changes in dietary intake between groups, per protocol (PP) analysis showed that the SUPP group increased protein (P < 0.01) and energy (P < 0.01) intakes. In contrast, protein and energy intakes further deteriorated in the CTRL group (PP). Although there was no difference in serum albumin and prealbumin changes between groups, in the total population serum albumin and prealbumin changes were positively associated with the increment in protein intake (r = 0.29, P = 0.01 and r = 0.27, P = 0.02, respectively). The SUPP group did not increase phosphate intake, phosphataemia remained unaffected, and the use of phosphate binders remained stable or decreased. The SUPP group exhibited improved SGA and QOL (P < 0.05). CONCLUSION: This study shows that providing maintenance haemodialysis patients with insufficient intake with a renal-specific oral supplement may prevent deterioration in nutritional indices and QOL without increasing the need for phosphate binders. PMID- 18408078 TI - Repression of gene expression by unphosphorylated NF-kappaB p65 through epigenetic mechanisms. AB - Cells from a "knock-in" mouse expressing a NF-kappaB p65 mutant bearing an alanine instead of serine at position 276 (S276A) display a significant reduction of NF-kappaB-dependent transcription, even though the mutant p65 forms appropriate complexes that translocate normally to the nucleus and bind to DNA. Surprisingly, however, instead of the expected embryonic lethality from hepatocyte apoptosis seen in the absence of NF-kappaB activity, the S276A knock in embryos die at different embryonic days due to variegated developmental abnormalities. We now demonstrate that this variegated phenotype is due to epigenetic repression resulting from the recruitment of histone deacetylases by the nonphosphorylatable form of NF-kappaB into the vicinity of genes positioned fortuitously near NF-kappaB-binding sites. Therefore, unphosphorylated nuclear NF kappaB can affect expression of genes not normally regulated by NF-kappaB through epigenetic mechanisms. PMID- 18408080 TI - VEGF-A, VEGFR-1, VEGFR-2 and Tie2 levels in plasma of premature infants: relationship to retinopathy of prematurity. AB - AIM: To study prospectively the plasma levels of vascular endothelial growth factor (VEGF-A), its soluble receptors sVEGFR-1, sVEGFR-2 and soluble Tie2 in premature infants. To identify their changes related to the onset of retinopathy of prematurity (ROP). METHODS: Blood samples of 63 preterm infants born at a postmenstrual age (PMA) of 23-32 weeks were obtained between 5 days and 15 weeks after birth. 42 infants had no ROP, two had stage 1, nine stage 2 and 10 stage 3. Of these, four infants were treated with retinal photocoagulation. VEGF-A, sVEGFR 1, sVEGFR-2, and sTie2 were measured in the plasma with a sandwich enzyme immunoassay using factor-specific monoclonal mouse antibodies. The time course of concentrations plotted by kernel smoothing in infants with and without ROP were compared and a paired subgroup with analysis of variance was analysed. RESULTS: ROP patients had raised plasma levels of sVEGFR-2 and sTie2 compared with premature infants without ROP. VEGF-A and sVEGFR-1 levels were similar in both groups. Analysis of a subgroup with pairs of measurements, one before 32 weeks and one after 36 weeks, showed a significant increase in sTie2 after 36 weeks of PMA independent of ROP (p = 0.03). CONCLUSION: This is the first study to measure plasma levels of angiogenic factors in ROP. Similar VEGF-A plasma levels in infants with and without ROP suggest that pathogenic retinal angiogenesis in ROP is mainly driven by local VEGF-A synthesis. Elevated plasma levels in active ROP were observed for sVEGFR-2 and sTie2. These increases have yet to be confirmed as predictive values for ROP. PMID- 18408081 TI - Is the maximum hypermetropic correction necessary in children with fully accommodative esotropia? AB - AIMS: This prospective study explores the effect of reduction in hypermetropic refractive correction on the angle and control of fully accommodative esotropia. METHODS: 30 childhood cases with fully accommodative esotropia were recruited. The angle of deviation with and without full hypermetropic correction (near and distance) was measured. The overall effect of reduction of the correction by one and two spherical dioptres (DS) on the angle and control of the deviation was identified. RESULTS: With the full hypermetropic correction in place, the angle of deviation for near was less than 10 prism dioptres (pd) in 73% of the participants, and the distance deviation was less than 10 pd in 93%. When the prescription was reduced by 1.00 DS, the percentage of those with a near deviation of less than 10 pd fell to 30% and 57% for the distance. Twenty per cent immediately decompensated to manifest esotropia with reduction of 1 dioptre of spectacle correction. CONCLUSION: Children with fully accommodative esotropia who are given the full hypermetropic correction demonstrate smaller, more controllable angles of deviation than those who are undercorrected by as little as only one dioptre. This supports the practice of providing the maximum hypermetropic correction for childhood esotropes. PMID- 18408079 TI - Mutations in String/CDC25 inhibit cell cycle re-entry and neurodegeneration in a Drosophila model of Ataxia telangiectasia. AB - Mutations in ATM (Ataxia telangiectasia mutated) result in Ataxia telangiectasia (A-T), a disorder characterized by progressive neurodegeneration. Despite advances in understanding how ATM signals cell cycle arrest, DNA repair, and apoptosis in response to DNA damage, it remains unclear why loss of ATM causes degeneration of post-mitotic neurons and why the neurological phenotype of ATM null individuals varies in severity. To address these issues, we generated a Drosophila model of A-T. RNAi knockdown of ATM in the eye caused progressive degeneration of adult neurons in the absence of exogenously induced DNA damage. Heterozygous mutations in select genes modified the neurodegeneration phenotype, suggesting that genetic background underlies variable neurodegeneration in A-T. The neuroprotective activity of ATM may be negatively regulated by deacetylation since mutations in a protein deacetylase gene, RPD3, suppressed neurodegeneration, and a human homolog of RPD3, histone deacetylase 2, bound ATM and abrogated ATM activation in cell culture. Moreover, knockdown of ATM in post mitotic neurons caused cell cycle re-entry, and heterozygous mutations in the cell cycle activator gene String/CDC25 inhibited cell cycle re-entry and neurodegeneration. Thus, we hypothesize that ATM performs a cell cycle checkpoint function to protect post-mitotic neurons from degeneration and that cell cycle re entry causes neurodegeneration in A-T. PMID- 18408082 TI - Use of multiplex PCR and real-time PCR to detect human herpes virus genome in ocular fluids of patients with uveitis. AB - AIM: To measure the genomic DNA of human herpes viruses (HHV) in the ocular fluids and to analyse the clinical relevance of HHV in uveitis. METHODS: After informed consent was obtained, a total of 111 ocular fluid samples (68 aqueous humour and 43 vitreous fluid samples) were collected from 100 patients with uveitis. The samples were assayed for HHV-DNA (HHV1-8) by using two different polymerase chain reaction (PCR) assays, qualitative PCR (multiplex PCR) and quantitative PCR (real-time PCR). RESULTS: In all of the patients with acute retinal necrosis (n = 16) that were tested, either the HSV1 (n = 2), HSV2 (n = 3), or VZV (n = 11) genome was detected. In all patients, high copy numbers of the viral DNA were also noted, indicating the presence of viral replication. In another 10 patients with anterior uveitis with iris atrophy, the VZV genome was detected. When using multiplex PCR, EBV-DNA was detected in 19 of 111 samples (17%). However, real-time PCR analysis of EBV-DNA indicated that there were only six of the 19 samples that had significantly high copy numbers. The cytomegalovirus (CMV) genome was detected in three patients with anterior uveitis of immunocompetent patients and in one immunocompromised CMV retinitis patient. In addition, one patient with severe unilateral panuveitis had a high copy number of HHV6-DNA. There was no HHV7- or HHV8-DNA detected in any of the samples. CONCLUSIONS: A qualitative multiplex PCR is useful in the screening of viral infections. However, the clinical relevance of the virus infection needs to be evaluated by quantitative real-time PCR. PMID- 18408083 TI - The use of infliximab in ocular inflammation. AB - AIMS: Experience with anti-tumour necrosis factor alpha medications in ophthalmology has been mainly in the treatment of resistant uveitis and scleritis. There have been a few case reports and one case series detailing the use of infliximab in the treatment of orbital inflammatory disease, but there is still limited experience with these agents in a variety of orbital inflammatory conditions. We describe successful suppression of inflammation with infliximab in the Idiopathic orbital inflammation, thyroid associated orbital inflammation and chronic relapsing inflammatory optic neuropathy. METHODS: This is a retrospective observational case series of five cases of orbital and ocular inflammation. RESULTS: We describe effects of infliximab in the treatment of three cases of orbital inflammation and two cases of chronic relapsing inflammatory optic neuropathy, all of which were unresponsive to alternative immune modulators. The patients received relief of symptoms and signs of inflammation in all five cases. However, this relief was not absolute as symptoms returned around week 5 post inflammation. CONCLUSION: This report highlights the expanding role of the biological agents in ophthalmology. Although not the answer in treatment, they offer a valuable alternative to steroids in cases of refractory inflammation. PMID- 18408084 TI - Can aortic MRI be used instead of transoesophagal echocardiography in patients with ischaemic stroke? PMID- 18408085 TI - Diffusion tensor imaging: implications for brain disease. PMID- 18408086 TI - Should we accept the status quo? Time for new trials in status epilepticus. PMID- 18408087 TI - White matter functional connectivity as an additional landmark for dominant temporal lobectomy. AB - Dominant temporal lobectomy is classically performed based on two criteria: a perfect knowledge of the temporo-mesial microsurgical anatomy and cortical landmarks laterally. However, the functional anatomy of the subcortical white matter tracts is taken into account less, despite the risk of inducing a permanent deficit (especially aphasia) if damaged. Even if Klinger's technique allows dissection of fibres on cadaveric specimens, the exact three dimensional geometry of these fasciculi remains poorly described. Tractography, based on diffusion tensor imaging (DTI), is a powerful tool to build three dimensional images of several fasciculi, helping neurosurgeons to create a mental representation of their relationships. Moreover, intraoperative subcortical electrostimulation enables mapping of the function of these pathways. Here we review the recent findings on the white matter anatomo-functional connectivity of the dominant temporal lobe, based on combined anatomical data provided by DTI and functional information provided by intraoperative stimulation. We then discuss their implications for temporal lobectomy, by using white matter functional connectivity as an additional landmark. PMID- 18408088 TI - Neurological picture. Multidetector-row computed tomography in the diagnosis of Collet-Sicard syndrome. PMID- 18408089 TI - Neurological picture. Varicella zoster virus and intracranial dolichoectasia in a late adult cancer survivor. PMID- 18408090 TI - Shoulder pain and external rotation in spastic hemiplegia do not improve by injection of botulinum toxin A into the subscapular muscle. AB - OBJECTIVE: To study the effect of botulinum toxin A in the subscapular muscle on shoulder pain and humerus external rotation. METHODS: 22 stroke patients with spastic hemiplegia, substantial shoulder pain and reduced external rotation of the humerus participated in a randomised, double blind, placebo controlled effect study. Injections of either botulinum toxin A (Botox, 2x50 units) or placebo were applied to the subscapular muscle at two locations. Pain was scored on a 100 mm vertical Visual Analogue Scale; external rotation was recorded by means of electronic goniometry. Assessments were carried out at 0 (baseline), 6 and 12 weeks. RESULTS: 21 patients completed the study. We observed no significant changes in pain or external rotation as a result of administration of botulinum toxin A. External rotation improved significantly (p = 0.001) for both the treatment group (20.4 degrees (16.6) to 32.1 degrees (14.0)) and the control group (10.3 degrees (19.5) to 23.7 degrees (20.7)) as a function of time. CONCLUSIONS: Application of botulinum toxin A into the subscapular muscle for reduction of shoulder pain and improvement of humeral external rotation in spastic hemiplegia does not appear to be clinically efficacious. PMID- 18408091 TI - Mutations of the SPG11 gene in patients with autosomal recessive spastic paraparesis and thin corpus callosum. PMID- 18408092 TI - Heat stress disorders and headache: a case of new daily persistent headache secondary to heat stroke. PMID- 18408093 TI - Successful repeated treatment with high dose cyclophosphamide and autologous blood stem cell transplantation in CIDP. PMID- 18408094 TI - Results of a UK questionnaire of diagnosis and treatment in cervical artery dissection. PMID- 18408096 TI - An infrequent cause of acute left lower quadrant abdominal pain. PMID- 18408097 TI - Hepatotoxicity of fast food? PMID- 18408098 TI - Nuclear factor-kappaB inhibition and non-alcoholic steatohepatitis: inflammation as a target for therapy. PMID- 18408099 TI - How does interferon inhibit HCV cell entry? PMID- 18408100 TI - Cobblestone appearance in the colon of a 24-year-old man. PMID- 18408101 TI - Amoebic liver abscess or is it? PMID- 18408102 TI - Pharmacological IKK2 inhibition blocks liver steatosis and initiation of non alcoholic steatohepatitis. AB - BACKGROUND: Non-alcoholic-steatohepatitis (NASH) leading to fibrosis, end-stage cirrhosis and hepatocellular carcinoma is an increasing health problem in the Western world. Thus, the need for new therapeutic approaches is increasing. IKK2 plays a key role in the development of NASH by mediating inflammation and insulin resistance. AIM: Here the beneficial effects of a pharmacological IKK2 inhibitor (AS602868) on initial stages of NASH progression were tested. METHODS: Mice were fed with a high sucrose diet (HSD) and daily-administered AS602868 and vehicle. The impact of AS602868 on NASH progression was studied using biochemical, histological and molecular markers. RESULTS: AS602868 treatment prevented HSD induced weight gain and visceral fat accumulation. In adipose tissue, AS602868 treated mice exhibited a lower degree of infiltrated macrophages along with reduced proinflammatory cytokine production. Further analysis demonstrated that AS602868 treatment efficiently inhibited nuclear factor (NF)-kappaB activation in liver non-parenchymal cells and as a consequence attenuated the inflammatory response in the liver. Accordingly, in HSD/AS602868 mice, liver and adipose tissue adiponectin levels remained at levels comparable with those of control chow-fed mice, while they were decreased in HSD/vehicle animals. Additionally, AS602868 improved lipid beta-oxidation mediated by peroxisome proliferator activated receptor (PPAR) alpha and PPARgamma. Systemic pharmacological IKK2 inhibition by AS602868 treatment efficiently prevented liver steatosis and inflammation, and improved antioxidant response. All this contributed to attenuation of NASH progression as evidenced by lower hepatocyte apoptosis and early stages of liver fibrosis. CONCLUSION: The data demonstrate that AS602868 mediated IKK2 inhibition represents a new therapeutic approach to prevent dietary induced NASH progression. PMID- 18408103 TI - Vomiting and a target sign on abdominal CT. PMID- 18408104 TI - Dysphagia and a skin rash. PMID- 18408105 TI - Gut, inflammation and osteoporosis: basic and clinical concepts. AB - Chronic inflammatory disorders such as inflammatory bowel diseases (IBD) affect bone metabolism and are frequently associated with the presence of osteoporosis. Bone loss is regulated by various mediators of the immune system such as the pro inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), IL-6, or interferon-gamma. TNF-alpha, a master cytokine in human IBD, causes bone erosions in experimental models and these effects are exerted by osteoclasts. Other TNF-related cytokines such as receptor activator of nuclear factor kappa B (RANK), its ligand, RANKL, and osteoprotegerin are important mediators in inflammatory processes in the gut and are critically involved in the pathophysiology of bone loss. The awareness and early diagnosis of osteoporosis in states of chronic inflammation, together with applied therapies such as bisphosphonates, may be beneficial in inflammation-associated osteoporosis. Although several mechanisms may contribute to osteoporosis in patients with IBD and coeliac disease, inflammation as an important factor has so far been neglected. As key inflammatory mediators in IBD such as TNF-alpha are involved in the disease process both in gut and bone, we hypothesise that neutralisation of TNF-alpha could prove an efficient strategy in the treatment of inflammation-related osteoporosis in the future. PMID- 18408106 TI - Ulcerative colitis following B lymphocyte depletion with rituximab in a patient with Graves' disease. PMID- 18408107 TI - Latent coeliac disease in childhood? PMID- 18408108 TI - Do we need treatment with tumour necrosis factor blockers for giant cell arteritis? PMID- 18408109 TI - Trials in rheumatoid arthritis: choosing the right outcome measure when minimal disease is achievable. PMID- 18408110 TI - Increased serum levels of soluble triggering receptor expressed on myeloid cells 1 in antineutrophil cytoplasmic antibody-associated vasculitis. PMID- 18408111 TI - Leflunomide as a remission-maintaining therapy in difficult-to-treat dermatomyositis. PMID- 18408112 TI - Association of anti-cyclic citrullinated peptide antibodies with subclinical atherosclerosis in patients with rheumatoid arthritis. PMID- 18408113 TI - Association of the toll-like receptor 4 gene polymorphisms with Behcet's disease. PMID- 18408114 TI - Expression of high mobility group protein 1 in the sera of patients and mice with systemic lupus erythematosus. PMID- 18408115 TI - Clinical and radiological picture of Jaccoud arthropathy in the context of systemic sclerosis. PMID- 18408116 TI - Uveitis and tumour necrosis factor blockade in ankylosing spondylitis. PMID- 18408117 TI - Is Sjogren myelopathy Devic disease? PMID- 18408119 TI - What's in a name? PMID- 18408120 TI - Process and impact evaluation of a legal assistance and health care community partnership. AB - Community health partnerships have increased in popularity, but their effectiveness is often not evaluated. Through secondary data analysis, this study evaluates a program that offered access to legal services to address health related issues, such as Medicaid reimbursement, Social Security benefits, medication coverage, and divorce. Based on the analysis reimbursements to expenditures, the health and law program appears to be cost-effective and thereby economically sustainable. The cost-effectiveness of this program increases the likelihood that it will be institutionalized and/or expanded. This program evaluation is used to exemplify how community stakeholders could partner to leverage resources to establish a sustainable community health and law program to address the needs of people living in medically underserved areas. PMID- 18408121 TI - Paracrine effects of hypoxic fibroblast-derived factors on the MPT-ROS threshold and viability of adult rat cardiac myocytes. AB - Cardiac fibroblasts contribute to multiple aspects of myocardial function and pathophysiology. The pathogenetic relevance of cytokine production by these cells under hypoxia, however, remains unexplored. With the use of an in vitro cell culture model, this study evaluated cytokine production by hypoxic cardiac fibroblasts and examined two distinct effects of hypoxic fibroblast-conditioned medium (HFCM) on cardiac myocytes and fibroblasts. Hypoxia caused a marked increase in the production of tumor necrosis factor (TNF)-alpha by cardiac fibroblasts. HFCM significantly enhanced the susceptibility of cardiac myocytes to reactive oxygen species (ROS)-induced mitochondrial permeability transition (MPT), determined by high-precision confocal line-scan imaging following controlled, photoexcitation-induced ROS production within individual mitochondria. Furthermore, exposure of cardiac myocytes to HFCM for 5 h led to loss of viability, as evidenced by change in morphology and annexin staining. HFCM also decreased DNA synthesis in cardiac fibroblasts. Normoxic fibroblast conditioned medium spiked with TNF-alpha at 200 pg/ml, a concentration comparable to that in HFCM, promoted loss of myocyte viability and decreased DNA synthesis in cardiac fibroblasts. These effects of HFCM are similar to the reported effects of hypoxia per se on these cell types, showing that hypoxic fibroblast-derived factors may amplify the distinct effects of hypoxia on cardiac cells. Importantly, because both hypoxia and oxidant stress prevail in a setting of ischemia and reperfusion, the effects of soluble factors from hypoxic fibroblasts on the MPT-ROS threshold and viability of myocytes may represent a novel paracrine mechanism that could exacerbate ischemia-reperfusion injury to cardiomyocytes. PMID- 18408122 TI - Mathematical model of the neonatal mouse ventricular action potential. AB - Therapies for heart disease are based largely on our understanding of the adult myocardium. The dramatic differences in action potential (AP) shape between neonatal and adult cardiac myocytes, however, indicate that a different set of molecular interactions in neonatal myocytes necessitates different treatment for newborns. Computational modeling is useful for synthesizing data to determine how interactions between components lead to systems-level behavior, but this technique has not been used extensively to study neonatal heart cell function. We created a mathematical model of the neonatal (day 1) mouse myocyte by modifying, on the basis of experimental data, the densities and/or formulations of ion transport mechanisms in an adult cell model. The new model reproduces the characteristic AP shape of neonatal cells, with a brief plateau phase and longer duration than the adult (action potential duration at 80% repolarization = 60.1 vs. 12.6 ms). The simulation results are consistent with experimental data, including 1) decreased density and altered inactivation of transient outward K+ currents, 2) increased delayed rectifier K+ currents, 3) Ca2+ entry through T type as well as L-type Ca2+ channels, 4) increased Ca2+ influx through Na+/Ca2+ exchange, and 5) Ca2+ transients resulting from transmembrane Ca2+ entry rather than release from the sarcoplasmic reticulum (SR). Simulations performed with the model generated novel predictions, including increased SR Ca2+ leak and elevated intracellular Na+ concentration in neonatal compared with adult myocytes. This new model can therefore be used for testing hypotheses and obtaining a better quantitative understanding of differences between neonatal and adult physiology. PMID- 18408123 TI - Brachial artery flow-mediated dilation during handgrip exercise: evidence for endothelial transduction of the mean shear stimulus. AB - Exercise elevates shear stress in the supplying conduit artery. Although this is the most relevant physiological stimulus for flow-mediated dilation (FMD), the fluctuating pattern of shear that occurs may influence the shear stress-FMD stimulus response relationship. This study tested the hypothesis that the brachial artery FMD response to a step increase in shear is influenced by the fluctuating characteristics of the stimulus, as evoked by forearm exercise. In 16 healthy subjects, we examined FMD responses to step increases in shear rate in three conditions: stable shear upstream of heat-induced forearm vasodilation (FHStable); fluctuating shear upstream of heat-induced forearm vasodilation and rhythmic forearm cuff inflation/deflation (FHFluctuating); and fluctuating shear upstream of exercise-induced forearm vasodilation (FEStep Increase). The mean increase in shear rate (+/-SD) was the same in all trials (FHFluctuating): 51.69 +/- 15.70 s(-1); FHStable: 52.16 +/- 14.10 s(-1); FEStep Increase: 50.14 +/- 13.03 s(-1) P = 0.131). However, the FHFluctuating and FEStep Increase trials resulted in a fluctuating shear stress stimulus with rhythmic high and low shear periods that were 96.18 +/- 24.54 and 11.80 +/- 7.30 s(-1), respectively. The initial phase of FMD (phase I) was followed by a second, delayed-onset FMD and was not different between conditions (phase I: FHFluctuating: 5.63 +/- 2.15%; FHStable: 5.33 +/- 1.85%; FEStep Increase: 5.30 +/- 2.03%; end-trial: FHFluctuating: 7.76 +/- 3.40%; FHStable: 7.00 +/- 3.03%; FEStep Increase: 6.68 +/ 3.04%; P = 0.196). Phase I speed also did not differ (P = 0.685). In conclusion, the endothelium transduced the mean shear when exposed to shear fluctuations created by a typical handgrip protocol. Muscle activation did not alter the FMD response. Forearm exercise may provide a viable technique to investigate brachial artery FMD in humans. PMID- 18408124 TI - Wave reflection augments central systolic and pulse pressures during facial cooling. AB - Cardiovascular events are more common in the winter months, possibly because of hemodynamic alterations in response to cold exposure. The purpose of this study was to determine the effect of acute facial cooling on central aortic pressure, arterial stiffness, and wave reflection. Twelve healthy subjects (age 23 +/- 3 yr; 6 men, 6 women) underwent supine measurements of carotid-femoral pulse wave velocity (PWV), brachial artery blood pressure, and central aortic pressure (via the synthesis of a central aortic pressure waveform by radial artery applanation tonometry and generalized transfer function) during a control trial (supine rest) and a facial cooling trial (0 degrees C gel pack). Aortic augmentation index (AI), an index of wave reflection, was calculated from the aortic pressure waveform. Measurements were made at baseline, 2 min, and 7 min during each trial. Facial cooling increased (P < 0.05) peripheral and central diastolic and systolic pressures. Central systolic pressure increased more than peripheral systolic pressure (22 +/- 3 vs. 15 +/- 2 mmHg; P < 0.05), resulting in decreased pulse pressure amplification ratio. Facial cooling resulted in a robust increase in AI and a modest increase in PWV (AI: -1.4 +/- 3.8 vs. 21.2 +/- 3.0 and 19.9 +/- 3.6%; PWV: 5.6 +/- 0.2 vs. 6.5 +/- 0.3 and 6.2 +/- 0.2 m/s; P < 0.05). Change in mean arterial pressure but not PWV predicted the change in AI, suggesting that facial cooling may increase AI independent of aortic PWV. Facial cooling and the resulting peripheral vasoconstriction are associated with an increase in wave reflection and augmentation of central systolic pressure, potentially explaining ischemia and cardiovascular events in the cold. PMID- 18408125 TI - Critical role of angiopoietins/Tie-2 in hyperglycemic exacerbation of myocardial infarction and impaired angiogenesis. AB - Angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) are the two ligands of the Tie 2 receptor, a receptor tyrosine kinase that is expressed on the endothelium. A balanced angiopoietin/Tie-2 system is critical for the maintenance of vascular integrity. We investigated the potential role of a disrupted angiopoietin/Tie-2 system on hyperglycemic exacerbation of myocardial infarction and impaired angiogenesis. Using streptozotocin (STZ) mice subjected to myocardial ischemia, we examined the effects of shifting the Ang-2-to-Ang-1 ratio on myocardial infarction size, apoptosis, bone marrow (BM) cell-endothelial progenitor cell (EPC) differentiation, and angiogenesis. In control mice, myocardial ischemia increased expression of both Ang-2 and Tie-2. In STZ mice, Ang-2 expression was elevated, whereas Tie-2 expression was reduced, and neither was significantly altered by ischemia. Myocardial infarct size and apoptosis were increased in STZ compared with control mice. Using in vivo administration of an adenovirus containing Ang-1 or Ang-2, we found that shifting the Ang-2-to-Ang-1 ratio to favor Ang-1 reduced myocardial apoptosis and infarct size in STZ mice, while shifting the Ang-2-to-Ang-1 ratio to favor Ang-2 resulted in a significant increase in myocardial infarct size and apoptosis in control mice. Myocardial ischemia-stimulated BM cell-EPC differentiation was inhibited and myocardial angiogenesis was reduced in STZ mice. Systemic administration of Ad-Ang-1 restored BM cell-EPC differentiation and increased myocardial VEGF expression and angiogenesis in STZ mice. Our data demonstrate that disturbed angiopoietin/Tie-2 signaling contributes to the hyperglycemic exacerbation of myocardial infarction and impaired angiogenesis. Restoration of the Ang-2-to-Ang-1 ratio may be a novel therapeutic strategy for the treatment of diabetic myocardial ischemic diseases. PMID- 18408126 TI - High-fat diet-induced obesity leads to increased NO sensitivity of rat coronary arterioles: role of soluble guanylate cyclase activation. AB - The impact of obesity on nitric oxide (NO)-mediated coronary microvascular responses is poorly understood. Thus NO-mediated vasomotor responses were investigated in pressurized coronary arterioles ( approximately 100 microm) isolated from lean (on normal diet) and obese (fed with 60% of saturated fat) rats. We found that dilations to acetylcholine (ACh) were not significantly different in obese and lean rats (lean, 83 +/- 4%; and obese, 85 +/- 3% at 1 microM), yet the inhibition of NO synthesis with N(omega)-nitro-l-arginine methyl ester reduced ACh-induced dilations only in vessels of lean controls. The presence of the soluble guanylate cyclase (sGC) inhibitor oxadiazolo-quinoxaline (ODQ) elicited a similar reduction in ACh-induced dilations in the two groups of vessels (lean, 60 +/- 11%; and obese, 57 +/- 3%). Dilations to NO donors, sodium nitroprusside (SNP), and diethylenetriamine (DETA)-NONOate were enhanced in coronary arterioles of obese compared with lean control rats (lean, 63 +/- 6% and 51 +/- 5%; and obese, 78 +/- 5% and 70 +/- 5%, respectively, at 1 microM), whereas dilations to 8-bromo-cGMP were not different in the two groups. In the presence of ODQ, both SNP and DETA-NONOate-induced dilations were reduced to a similar level in lean and obese rats. Moreover, SNP-stimulated cGMP immunoreactivity in coronary arterioles and also cGMP levels in carotid arteries were enhanced in obese rats, whereas the protein expression of endothelial NOS and the sGC beta1-subunit were not different in the two groups. Collectively, these findings suggest that in coronary arterioles of obese rats, the increased activity of sGC leads to an enhanced sensitivity to NO, which may contribute to the maintenance of NO-mediated dilations and coronary perfusion in obesity. PMID- 18408127 TI - CD4+ T lymphocytes mediate hypercholesterolemia-induced endothelial dysfunction via a NAD(P)H oxidase-dependent mechanism. AB - Although hypercholesterolemia is known to impair endothelium-dependent vasodilation (EDV) long before the appearance of atherosclerotic plaques, it remains unclear whether the immune mechanisms that have been implicated in atherogenesis also contribute to the early oxidative stress and endothelial cell dysfunction elicited by hypercholesterolemia. EDV (wire myography), superoxide generation (cytochrome c reduction), and NAD(P)H oxidase mRNA expression were monitored in aortic rings from wild-type (WT) and mutant mice placed on either a normal diet or a cholesterol-enriched diet (HC) for 2 wk. WT mice on HC exhibited impaired EDV, enhanced superoxide generation, and increased expression of NAD(P)H oxidase subunit Nox-2 mRNA. The impaired EDV and increased superoxide generation induced by HC were significantly blunted in severe combined immunodeficient (SCID) mice and CD4+ T lymphocyte-deficient mice. These responses were also attenuated in HC mice genetically deficient in IFN-gamma; however, adoptive transfer of WT-HC CD4+ T lymphocytes to IFN-gamma-deficient recipients restored HC-induced responses. The HC-induced impaired EDV and oxidative stress were also attenuated in HC mice genetically deficient in Nox-2 (gp91(phox-/-)) and in WT- >gp91(phox-/-)-HC chimeras. HC-induced gp91(phox) mRNA expression was significantly blunted in mice deficient in CD4+ T cells or IFN-gamma and was restored with adoptive transfer of WT-HC CD4+ T cells to IFN-gamma-deficient recipients. These findings implicate the immune system in the early endothelial cell dysfunction associated with hypercholesterolemia and are consistent with a mechanism of impaired EDV that is mediated by CD4+ T cells and IFN-gamma, acting through the generation of superoxide from vascular NAD(P)H oxidase. PMID- 18408128 TI - Chronic beta2-adrenoceptor stimulation impairs cardiac relaxation via reduced SR Ca2+-ATPase protein and activity. AB - We determined the cardiovascular effects of chronic beta2-adrenoceptor (beta2-AR) stimulation in vivo and examined the mechanism for the previously observed prolonged diastolic relaxation. Rats (3 mo old; n = 6), instrumented with implantable radiotelemeters, received the selective beta2-AR agonist formoterol (25 microg.kg(-1).day(-1) ip) for 4 wk, with selected cardiovascular parameters measured daily throughout this period, and for a further 7 days after cessation of treatment. Chronic beta2-AR stimulation was associated with an increase in heart rate (HR) of 17% (days 1-14) and 5% (days 15-28); a 11% (days 1-14) and 6% (days 15-28) decrease in mean arterial blood pressure; and a 24% (days 1-14) increase in the rate of cardiac relaxation (-dP/dt) compared with initial values (P < 0.05). Cessation of beta2-AR stimulation resulted in an 8% decrease in HR and a 7% decrease in -dP/dt, compared with initial values (P < 0.05). The prolonged cardiac relaxation with chronic beta2-AR stimulation was associated with a 30% decrease in the maximal rate (Vmax) of sarco(endo)plasmic reticulum Ca2+ -ATPase (SERCA) activity, likely attributed to a 50% decrease in SERCA2a protein (P < 0.05). glycogen synthase kinase-3beta (GSK-3beta) has been implicated as a negative regulator of SERCA2 gene transcription, and we observed a approximately 60% decrease (P < 0.05) in phosphorylated GSK-3beta protein after chronic beta2-AR stimulation. Finally, we found a 40% decrease (P < 0.05) in the mRNA expression of the novel A kinase anchoring protein AKAP18, also implicated in beta2-AR-mediated cardiac relaxation. These findings highlight some detrimental cardiovascular effects of chronic beta2-AR agonist administration and identify concerns for their current and future use for treating asthma or for conditions where muscle wasting and weakness are indicated. PMID- 18408129 TI - Role of glucose metabolism in the recovery of postischemic LV mechanical function: effects of insulin and other metabolic modulators. AB - The role of proton (H+) production from glucose metabolism in the recovery of myocardial function during postischemic reperfusion and its alteration by insulin and other metabolic modulators were examined. Rat hearts were perfused in vitro with Krebs-Henseleit solution containing palmitate (1.2 mmol/l) and glucose (11 mmol/l) under nonischemic conditions or during reperfusion following no-flow ischemia. Perfusate contained normal insulin (n-Ins, 50 mU/l), zero insulin (0 Ins), or supplemental insulin (s-Ins, 1,000 mU/l) or other metabolic modulators [dichloroacetate (DCA) at 3 mmol/l, oxfenicine at 1 mmol/l, and N6 cyclohexyladenosine (CHA) at 0.5 micromol/l]. Relative to n-Ins, 0-Ins depressed rates of glycolysis and glucose oxidation in nonischemic hearts and impaired recovery of postischemic function. Relative to n-Ins, s-Ins did not affect aerobic glucose metabolism and did not improve recovery when present during reperfusion. When present during ischemia and reperfusion, s-Ins impaired recovery. Combinations of metabolic modulators with s-Ins stimulated glucose oxidation approximately 2.5-fold in nonischemic hearts and reduced H+ production. DCA and CHA, in combination with s-Ins, improved recovery of function, but addition of oxfenicine to this combination provided no further benefit. Although DCA and CHA were each partially protective in hearts perfused with n-Ins, optimal protection was achieved with DCA + CHA; recovery of function was inversely proportional to H+ production during reperfusion. Although supplemental insulin is not beneficial, elimination of H+ production from glucose metabolism by simultaneous inhibition of glycolysis and stimulation of glucose oxidation optimizes recovery of postischemic mechanical function. PMID- 18408130 TI - VEGF promotes vascular sympathetic innervation. AB - The sympathetic nervous system, via postganglionic innervation of blood vessels and the heart, is an important determinant of cardiovascular function. The mechanisms underlying sympathetic innervation of targets are not fully understood. This study tests the hypothesis that target-derived vascular endothelial growth factor (VEGF) promotes sympathetic innervation of blood vessels. Western blot and immunohistochemical analyses indicate that VEGF is produced by vascular cells in arteries and that VEGF receptors are expressed on sympathetic nerve fibers innervating arteries. In vitro, exogenously added VEGF and VEGF produced by vascular smooth muscle cells (VSMCs) in sympathetic neurovascular cocultures inhibited semaphorin 3A (Sema3A)-induced collapse of sympathetic growth cones. In the absence of Sema3A, VEGF and VSMCs also increased growth cone area. These effects were mediated via VEGF receptor 1. In vivo, the neutralization of VEGF inhibited the reinnervation of denervated femoral arteries. These data demonstrate that target-derived VEGF plays a previously unrecognized role in promoting the growth of sympathetic axons. PMID- 18408131 TI - Angiotensin II receptor antagonism reverts the selective cardiac BNP upregulation and secretion observed in myocarditis. AB - The cardiac natriuretic peptides atrial natriuretic factor (ANF) and brain natriuretic peptide (BNP) are discoordinately regulated in myocardial inflammation associated with acute allograft rejection in humans and during in vitro exposure of cardiocyte cultures to some proinflammatory cytokines. We used experimental autoimmune myocarditis (EAM) to determine whether the discoordinate regulation of ANF and BNP was specific to the situations above or was generally associated with other types of myocardial inflammation. The dependency of this process to angiotensin signaling was also determined, given that previous work demonstrated beneficial effects of the angiotensin receptor blocker olmesartan in myocarditis. Histopathological changes, plasma and cardiac ANF, BNP, and selected cytokines gene expression as well as plasma cytokine levels using a cytokine array were determined in EAM, angiotensin receptor blocker-treated, and control rats. It was found that EAM specifically increases BNP but not ANF circulating levels, thus mimicking the findings in acute cardiac allograft rejection and the effect of some proinflammatory cytokines on cardiocyte cultures in vitro. Plasma cytokine array and real-time PCR revealed that lipopolysaccharide-induced CXC chemokine, monocyte chemotactic protein-1, and tissue inhibitor of metalloproteinase-1 were increased in plasma and in the myocardium of EAM rats. Olmesartan treatment reversed virtually all neuroendocrine and histopathological cardiac changes induced by EAM, thus providing a mechanistic insight into this phenomenon. It is concluded that the inflammatory process contributes specific cytokines, leading to the disregulation of cardiac ANF and BNP production observed during myocardial inflammation, and that this process is angiotensin receptor 1 dependent. PMID- 18408132 TI - Localization of the novel angiotensin peptide, angiotensin-(1-12), in heart and kidney of hypertensive and normotensive rats. AB - A low expression of angiotensinogen in the heart has been construed as indicating a circulating uptake mechanism to explain the local effects of angiotensin II on tissues. The recent identification of angiotensin-(1-12) in an array of rat organs suggests this propeptide may be an alternate substrate for local angiotensin production. To test this hypothesis, tissues from 11-wk-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats (n = 14) were stained with purified antibodies directed to the COOH terminus of angiotensin-(1 12). Robust angiotensin-(1-12) staining was predominantly found in ventricular myocytes with less staining found in the medial layer of intracoronary arteries and vascular endothelium. In addition, angiotensin-(1-12) immunoreactivity was present in the proximal, distal, and collecting renal tubules within the deep cortical and outer medullary zones in both strains. Preadsorption of the antibody with angiotensin-(1-12) abolished staining in both tissues. Corresponding tissue measurements by radioimmunoassay showed 47% higher levels of angiotensin-(1-12) in the heart of SHR compared with WKY rats (P < 0.05). In contrast, renal angiotensin-(1-12) levels were 16.5% lower in SHR compared with the WKY rats (P < 0.05). This study shows for first time the localization of angiotensin-(1-12) in both cardiac myocytes and renal tubular components of WKY and SHR. In addition, we show that increased cardiac angiotensin-(1-12) concentrations in SHR is associated with a small, but statistically significant, reduction in renal angiotensin-(1-12) levels. PMID- 18408134 TI - Incidence of protein on actin bridges between endothelium and smooth muscle in arterioles demonstrates heterogeneous connexin expression and phosphorylation. AB - Although much physiology in resistance vessels has been attributed to the cytoplasmic connection between endothelial cells (ECs) and vascular smooth muscle cells (VSMCs), little is known of the protein expression between the two cell types. In an attempt to identify the proteins between ECs and VSMCs, mouse cremaster arterioles were stained with phalloidin-Alexa 594 and viewed on a confocal microscope that resolved "actin bridges" within the internal elastic lamina between ECs and VSMCs. To determine the incidence of protein, the pixel intensity from the antibodies on actin bridges were compared with the pixel intensity from antibodies within ECs or VSMCs. N-cadherin, desmin, connexin (Cx)40, and Cx43 and phosphorylated Cx43 at serine-368 were identified on actin bridges, but NG2, CD31, and Cx45 were not evident. Cx37 expression was more variable than the other connexins examined. Using this method on rat mesentery, we confirm the previously published predominance of Cx37 and Cx40 at the myoendothelial junction that was determined using electron microscopy. We conclude that this new method represents an important screening mechanism in which to rapidly test for protein expression between ECs and VSMCs and possibly a first-step in quantifying protein expression at the myoendothelial junction. PMID- 18408133 TI - Impaired relaxation is the main manifestation in transgenic mice expressing a restrictive cardiomyopathy mutation, R193H, in cardiac TnI. AB - Transgenic mice were generated to express a restrictive cardiomyopathy (RCM) human cardiac troponin I (cTnI) R192H mutation in the heart (cTnI(193His) mice). The objective of this study was to assess cardiac function during the development of diastolic dysfunction and to gain insight into the pathophysiological impact of the RCM cTnI mutation. Cardiac function and pathophysiological changes were monitored in cTnI193His mice and wild-type littermates for a period of 12 mo. It progressed gradually from abnormal relaxation to diastolic dysfunction characterized with high-resolution echocardiography by a reversed E-to-A ratio, increased deceleration time, and prolonged isovolumetric relaxation time. At the age of 12 mo, cardiac output in cTnI(193His) mice was significantly declined, and some transgenic mice showed congestive heart failure. The negative impact of cTnI193His on ventricular contraction and relaxation was further demonstrated in isolated mouse working heart preparations. The main morphological change in cTnI193His myocytes was shortened cell length. Dobutamine stimulation increased heart rate in cTnI193His mice but did not improve CO. The cTnI193His mice had a phenotype similar to that in human RCM patients carrying the cTnI mutation characterized morphologically by enlarged atria and restricted ventricles and functionally by diastolic dysfunction and diastolic heart failure. The results demonstrate a critical role of the COOH-terminal domain of cTnI in the diastolic function of cardiac muscle. PMID- 18408135 TI - Differential loss of cytochrome-c oxidase subunits in ischemia-reperfusion injury: exacerbation of COI subunit loss by PKC-epsilon inhibition. AB - We have previously described a PKC-epsilon interaction with cytochrome oxidase subunit IV (COIV) that correlates with enhanced CO activity and cardiac ischemic preconditioning (PC). We therefore investigated the effects of PC and ischemia reperfusion (I/R) injury on CO subunit levels in an anesthetized rat coronary ligation model. Homogenates prepared from the left ventricular regions at risk (RAR) and not at risk (RNAR) for I/R injury were fractionated into cell-soluble (S), 600 g low-speed centrifugation (L), gradient-purified mitochondrial (M), and 100,000 g particulate (P) fractions. In RAR tissue, PC (2 cycles of 5-min ischemia and 5-min reperfusion) decreased the COI in the P fraction ( approximately 29% of total cellular COI), suggesting changes in interfibrillar mitochondria. After 30 min of ischemia and 120 min of reperfusion, total COI levels decreased in the RAR by 72%. Subunit Va was also downregulated by 42% following prolonged I/R in the RAR. PC administered before I/R reduced the loss of COI in the M and P fractions approximately 30% and prevented COVa losses completely. We observed no losses in subunits Vb and VIIa following I/R alone; however, significant losses occurred when PC was administered before prolonged I/R. Delivery of a cell-permeable PKC-epsilon translocation inhibitor (epsilonV1 2) to isolated rat hearts before prolonged I/R dramatically increased COI loss, suggesting that PKC-epsilon protects COI levels. We propose that additional measures to protect CO subunits when coadministered with PC may improve its cardioprotection against I/R injury. PMID- 18408136 TI - p38 MAPK-dependent eNOS upregulation is critical for 17beta-estradiol-mediated cardioprotection following trauma-hemorrhage. AB - Studies have shown that p38 MAPK and nitric oxide (NO), generated by endothelial NO synthase (eNOS), play key roles under physiological and pathophysiological conditions. Although administration of 17beta-estradiol (E2) protects cardiovascular injury from trauma-hemorrhage, the mechanism by which E2 produces those effects remains unknown. Our objective was to determine whether the E2 mediated activation of myocardial p38 MAPK and subsequent eNOS expression/phosphorylation would protect the heart following trauma-hemorrhage. To study this, male Sprague-Dawley rats underwent soft-tissue trauma (midline laparatomy) and hemorrhagic shock (mean blood pressure 35-40 mmHg for 90 min), followed by fluid resuscitation. Animals were pretreated with specific p38 MAPK inhibitor SB-203580 (SB; 2 mg/kg), and nonselective NO synthase inhibitor NG nitro-l-arginine methyl ester (l-NAME; 30 mg/kg) 30 min before vehicle (cyclodextrin) or E2 (100 microg/kg) treatment, followed by resuscitation, and were killed 2 h thereafter. Cardiovascular performance and other parameters were measured. E2 administration following trauma-hemorrhage increased cardiac p38 MAPK activity, eNOS expression and phosphorylation at Ser(1177), and nitrate/nitrite levels in plasma and heart tissues; these were associated with normalized cardiac performance, which was reversed by SB administration. In addition, E2 also prevented trauma-hemorrhage-induced increase in cytokines (IL-6 and TNF-alpha), chemokines (macrophage inflammatory protein-2 and cytokine induced neutrophil chemoattractant-1), and ICAM-1, which was reversed by l-NAME administration. Administration of E2 following trauma-hemorrhage attenuated cardiac tissue injury markers, myeloperoxidase activity, and nitrotyrosine level, which were reversed by treatment with SB and l-NAME. The salutary effects of E2 on cardiac functions and tissue protection following trauma-hemorrhage are mediated, in part, through activation of p38 MAPK and subsequent eNOS expression and phosphorylation. PMID- 18408137 TI - The pig as an experimental model for elucidating the mechanisms governing dietary influence on mineral absorption. AB - This review highlights the similarities between pigs and humans and thereby the value of the porcine human nutritional model, and reviews some of the more recent applications of this model for nutritional research. PMID- 18408138 TI - Generation of mature dendritic cells with unique phenotype and function by in vitro short-term culture of human monocytes in the presence of interleukin-4 and interferon-beta. AB - Dendritic cell (DC)-based immunotherapy has been utilized for the treatment of not only a number of human malignancies but also a select group of infectious diseases. Conventional techniques for the generation and maturation of DCs require 7 days of in vitro culture, which prompted us to seek alternative methods that would hasten the generation of functional human myeloid DCs in vitro. Following the use of a number of cytokines/growth factors, we found that in vitro culture of purified human monocytes, in media containing interleukin (IL)-4, together with interferon (IFN)-beta for 24 hrs, followed by the addition of non specific antigenic stimuli, such as keyhole limpet hemocyanin (KLH), lipopolysaccharide (LPS), or inactivated human immunodeficiency virus (HIV)-1 induced the monocytes to differentiated by 3 days into mature DCs (4B-DCs). These 4B-DCs expressed high levels of CD83 and CD11c, as well as markers of immune activation, including CD80 and CD86, human leukocyte antigen (HLA) class I and II, and CD14, but not CD1a. Anti-CD14 blocking antibody interfered with generation of 4B-DCs by LPS, but not by KLH or HIV-1. Interestingly, 4B-DCs, but not conventional DCs generated using macrophage-colony stimulating factor and IL 4 (G4-DCs), expressed OX40 and OX40L. 4B-DCs showed phagocytic activity, and spontaneously produced IL-12 and tumor necrosis factor (TNF)-alpha, but not IL 10. 4B-DCs promoted proliferation of allogeneic naive CD4(+) T cells, producing IFN-(lambda) at lower levels than those stimulated with G4-DCs. 4B-DCs were more potent stimulators of allogeneic bulk CD8(+) T cells producing IFN-(lambda) than G4-DCs. These data indicate that 4B-DCs are unique and may provide a relatively more rapid alternative tool for potential clinical use, as compared with conventional G4-DCs. PMID- 18408139 TI - Chondroitin-4-sulphate reduced oxidative injury in caerulein-induced pancreatitis in mice: the involvement of NF-kappaB translocation and apoptosis activation. AB - Activation of nuclear factor kappaB (NF-kappaB) and caspases may greatly amplify inflammation and cell damage in addition to that directly exerted by free radicals. Since reactive oxygen species (ROS) are involved in acute pancreatitis, we studied whether the administration of chondroitin-4-sulphate (C4S), in addition to its antioxidant activity, was able to modulate NF-kappaB and caspase activation in an experimental model of caerulein-induced acute pancreatitis in mice. Hyperstimulating doses of caerulein (50 microg/ kg), five injections per mouse given at hourly intervals produced the following: high serum lipase and amylase activity; lipid peroxidation, evaluated by 8-isoprostane concentrations; loss of antioxidant defenses such as glutathione reductase (GR) activity; NF kappaB activation and loss of cytoplasmic IkappaBalpha protein; increases in tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), caspase-3, and caspase-7 gene expression and their related protein; accumulation and activation of neutrophils in the damaged tissue, evaluated by elastase (ELA) determination; and pancreatic injury, evaluated by histologic analysis. Pretreatment of mice with different doses of C4S, given 1 hr before caerulein injections and 1 and 2 hrs after the last caerulein injection, reduced lipid peroxidation, inhibited NF kappaB translocation and cytoplasmic IkappaBalpha protein loss, decreased TNF alpha, IL-6, and caspase gene expression and their related protein levels, limited endogenous antioxidant depletion, and reduced tissue neutrophils accumulation and tissue damage. Since molecules with antioxidant activity can block NF-kappaB and apoptosis activation, we suggest that C4S administration is able to block NF-kappaB and caspase activation by reducing the oxidative burst. PMID- 18408140 TI - The importance of the omega-6/omega-3 fatty acid ratio in cardiovascular disease and other chronic diseases. AB - Several sources of information suggest that human beings evolved on a diet with a ratio of omega-6 to omega-3 essential fatty acids (EFA) of approximately 1 whereas in Western diets the ratio is 15/1-16.7/1. Western diets are deficient in omega-3 fatty acids, and have excessive amounts of omega-6 fatty acids compared with the diet on which human beings evolved and their genetic patterns were established. Excessive amounts of omega-6 polyunsaturated fatty acids (PUFA) and a very high omega-6/omega-3 ratio, as is found in today's Western diets, promote the pathogenesis of many diseases, including cardiovascular disease, cancer, and inflammatory and autoimmune diseases, whereas increased levels of omega-3 PUFA (a lower omega-6/omega-3 ratio), exert suppressive effects. In the secondary prevention of cardiovascular disease, a ratio of 4/1 was associated with a 70% decrease in total mortality. A ratio of 2.5/1 reduced rectal cell proliferation in patients with colorectal cancer, whereas a ratio of 4/1 with the same amount of omega-3 PUFA had no effect. The lower omega-6/omega-3 ratio in women with breast cancer was associated with decreased risk. A ratio of 2-3/1 suppressed inflammation in patients with rheumatoid arthritis, and a ratio of 5/1 had a beneficial effect on patients with asthma, whereas a ratio of 10/1 had adverse consequences. These studies indicate that the optimal ratio may vary with the disease under consideration. This is consistent with the fact that chronic diseases are multigenic and multifactorial. Therefore, it is quite possible that the therapeutic dose of omega-3 fatty acids will depend on the degree of severity of disease resulting from the genetic predisposition. A lower ratio of omega 6/omega-3 fatty acids is more desirable in reducing the risk of many of the chronic diseases of high prevalence in Western societies, as well as in the developing countries. PMID- 18408141 TI - Identification of proteins secreted from leptin stimulated MCF-7 breast cancer cells: a dual proteomic approach. AB - Leptin is an adipocyte-derived hormone that regulates energy expenditure and food intake. A significant role for leptin in breast cancer has also been indicated by the resistance of leptin knockout mice in development of mammary tumors. In vitro, leptin induces proliferation of MCF-7 cells by activating cellular signaling pathways (1, 11, 12, 16, 17, 56). As leptin is emerging as an important factor for tumor growth, and hormones can exert their actions via autocrine/paracrine mechanisms, we hypothesized leptin may act by regulating epithelial-derived proteins. To test this hypothesis, leptin-regulated proteins secreted from MCF-7 mammary tumor cells were identified using proteomics techniques. Treatment of MCF-7 cells with 500 ng/ml leptin for 24 hours resulted in a 40% increase in cell number and a 5-fold increase in protein secretion as compared to controls. Establishing the significance of leptin-induced secreted factors, the addition of conditioned media from leptin-treated MCF-7 cells to synchronized MCF-7 cells resulted in 40% increase in cell number. Identification of leptin-regulated secreted proteins was done by 2D gel electrophoresis coupled with MALDI-TOF mass spectrometry. Proteins identified using Pro Found software and NCBI database included KF10 Collagen Precursor, Serologically Defined Breast Cancer Antigen NY-BR-62 and Cortactin Isoform a. A Human Cytokine Antibody Array system was used to identify low abundant proteins in the media of control and 500 ng/ml leptin-stimulated MCF-7 cells. In leptin treated cells, levels of FGF-9 were increased while IGFBP-3 and TGF-beta3 levels were decreased. Many previous studies have focused on the regulation of distinct cellular proteins by leptin during mammary tumor cell proliferation. However, ours is the first study to identify leptin-regulated secreted proteins, many of which are known to play important roles in cancer. Our data support that leptin can influence mammary tumor growth and progression through regulation of autocrine/paracrine factors and by modulating the extracellular matrix composition. PMID- 18408142 TI - Hypoxic conditioning suppresses nitric oxide production upon myocardial reperfusion. AB - Physiologically modulated concentrations of nitric oxide (NO) are generally beneficial, but excessive NO can injure myocardium by producing cytotoxic peroxynitrite. Recently we reported that intermittent, normobaric hypoxia conditioning (IHC) produced robust cardioprotection against infarction and lethal arrhythmias in a canine model of coronary occlusion-reperfusion. This study tested the hypothesis that IHC suppresses myocardial nitric oxide synthase (NOS) activity and thereby dampens explosive, excessive NO formation upon reperfusion of occluded coronary arteries. Mongrel dogs were conditioned by a 20 d program of IHC (FIO(2) 9.5-10%; 5-10 min hypoxia/cycle, 5-8 cycles/d with intervening 4 min normoxia). One day later, ventricular myocardium was sampled for NOS activity assays, and immunoblot detection of the endothelial NOS isoform (eNOS). In separate experiments, myocardial nitrite (NO(2)(-)) release, an index of NO formation, was measured at baseline and during reperfusion following 1 h occlusion of the left anterior descending coronary artery (LAD). Values in IHC dogs were compared with respective values in non-conditioned, control dogs. IHC lowered left and right ventricular NOS activities by 60%, from 100-115 to 40-45 mU/g protein (P < 0.01), and decreased eNOS content by 30% (P < 0.05). IHC dampened cumulative NO(2)(-) release during the first 5 min reperfusion from 32 +/- 7 to 14 +/- 2 mumol/g (P < 0.05), but did not alter hyperemic LAD flow (15 +/ 2 vs. 13 +/- 2 ml/g). Thus, IHC suppressed myocardial NOS activity, eNOS content, and excessive NO formation upon reperfusion without compromising reactive hyperemia. Attenuation of the NOS/NO system may contribute to IHC induced protection of myocardium from ischemia-reperfusion injury. PMID- 18408143 TI - Coordinated upregulation of a series of endogenous antioxidants and phase 2 enzymes as a novel strategy for protecting renal tubular cells from oxidative and electrophilic stress. AB - In view of the crucial involvement of oxidative and electrophilic stress in various kidney disorders, this study was undertaken to test the hypothesis that pharmacologically-mediated coordinated upregulation of endogenous renal antioxidants and phase 2 enzymes is an effective strategy for renal protection. Notably, studies on the pharmacological inducibility of a series of antioxidants and phase 2 enzymes in renal tubular cells are lacking. Here we reported that incubation of normal rat kidney (NRK-52E) proximal tubular cells with low micromolar concentrations (10-50 microM) of the cruciferous nutraceutical, 1,2 dithiole-3-thione (D3T), led to a significant concentration-dependent induction of a wide spectrum of antioxidants and phase 2 enzymes, including catalase (CAT), reduced form of glutathione (GSH), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), NAD(P)H:quinone oxidoreductase 1 (NQO1), and heme oxygenase (HO). We further observed that D3T treatment also increased the protein and mRNA expression for CAT, gamma-glutamylcysteine ligase, GR, GST-A, GST-M, NQO1, and HO-1. Incubation of the renal tubular cells with H(2)O(2), SIN-1-derived peroxynitrite, or 4-hydroxy-2-nonenal led to concentration-dependent decreases in cell viability. Pretreatment of the renal tubular cells with 10-50 microM D3T afforded remarkable protection against the nephrocytotoxicity elicited by the above oxidative and electrophilic species. The D3T-mediated cytoprotection showed a concentration-dependent relationship. Taken together, this study for the first time comprehensively characterized the inducibility by a unique nutraceutical of a wide spectrum of antioxidative and phase 2 defenses in renal tubular cells at the levels of enzyme activity as well as protein and mRNA expression, and demonstrated that such a coordinated upregulation of cellular defenses led to remarkable protection of renal tubular cell from oxidative and electrophilic stress. Because of the crucial role of oxidative and electrophilic stress in inflammatory injury, D3T-mediated coordinated induction of endogenous antioxidative and phase 2 defenses may also serve as an important anti-inflammatory mechanism in kidneys. PMID- 18408144 TI - Effect of Buddleja officinalis on high-glucose-induced vascular inflammation in human umbilical vein endothelial cells. AB - In this study, we aimed to investigate whether an aqueous extract of Buddleja officinalis (ABO) suppresses high-glucose-induced vascular inflammatory processes in the primary cultured human umbilical vein endothelial cells (HUVEC). The high glucose-induced increase in expression of cell adhesion molecules (CAMs) such as intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial-selectin (E-selectin) was significantly attenuated by pretreatment with ABO in a dose-dependent manner. Enhanced cell adhesion caused by high glucose in co-cultured U937 and HUVEC was also blocked by pretreatment with ABO. Pretreatment with ABO also blocked formation of high-glucose-induced reactive oxygen species (ROS). In addition, ABO suppressed the transcriptional activity of NF-kappaB and IkappaB phosphorylation under high-glucose conditions. Pretreatment with N(G)-nitro-l-arginine methyl ester (L-NAME), an endothelial nitric oxide (NO) synthase inhibitor, attenuated the protective action of ABO on high-glucose-induced CAM expression, suggesting a potential role of NO signaling. The present data suggest that ABO could suppress high-glucose-induced vascular inflammatory processes, and ABO may be closely related with the inhibition of ROS and NF-kappaB activation in HUVEC. PMID- 18408145 TI - Intermittent hypoxia: cause of or therapy for systemic hypertension? AB - During acute episodes of hypoxia, chemoreceptor-mediated sympathetic activity increases heart rate, cardiac output, peripheral resistance and systemic arterial pressure. However, different intermittent hypoxia paradigms produce remarkably divergent effects on systemic arterial pressure in the post-hypoxic steady state. The hypertensive effects of obstructive sleep apnea (OSA) vs. the depressor effects of therapeutic hypoxia exemplify this divergence. OSA, a condition afflicting 15-25% of American men and 5-10% of women, has been implicated in the pathogenesis of systemic hypertension and is a major risk factor for heart disease and stroke. OSA imposes a series of brief, intense episodes of hypoxia and hypercapnia, leading to persistent, maladaptive chemoreflex-mediated activation of the sympathetic nervous system which culminates in hypertension. Conversely, extensive evidence in animals and humans has shown controlled intermittent hypoxia conditioning programs to be safe, efficacious modalities for prevention and treatment of hypertension. This article reviews the pertinent literature in an attempt to reconcile the divergent effects of intermittent hypoxia therapy and obstructive sleep apnea on hypertension. Special emphasis is placed on research conducted in the nations of the former Soviet Union, where intermittent hypoxia conditioning programs are being applied therapeutically to treat hypertension in patients. Also reviewed is evidence regarding mechanisms of the pro- and anti-hypertensive effects of intermittent hypoxia. PMID- 18408146 TI - Expression of alpha-actinin-1 in human glomerular mesangial cells in vivo and in vitro. AB - Recent studies have demonstrated important roles of alpha-actinins in glomerular disease, while little information is known about the expression profile of alpha actinins in human glomerular mesangial cells. Here, immunofluorescence and confocal microscopy showed that alpha-actinin-1 exclusively distributed along mesangial cells in human glomeruli of IgA nephropathy. RT-PCR and Western blot further confirmed the expression of alpha-actinin-1 in primary cultured human mesangial cells. We also found that transforming growth factor-beta 1 (TGF-beta 1) stimulated ACTN1 gene transcription and that transiently transfected alpha actinin-1 significantly increased TGF-beta 1-induced plasminogen activator inhibitor-1 (PAI-1) promoter activity in human mesangial cells. These findings suggest that alpha-actinin-1 may play a role in human glomerular disease. PMID- 18408147 TI - Delipidated retroviruses as potential autologous therapeutic vaccines--a pilot experiment. AB - This pilot experiment in a simian immunodeficiency virus (SIV) chronic infection model aimed at extending our previous findings that vaccination with delipidated SIV resulted in more potent and diversified antiviral responses (1). Macaques chronically infected with SIVmac239 treated with antiretroviral therapy (ART) were vaccinated with autologous delipidated virus via consecutive lymph node targeted immunizations-1, 1 and 10 mug of virus spaced monthly. Results showed all animals had lasting viral load reduction approaching 1 log compared to set point, and disease delay. Delipidation may enhance processing/ presentation of viral antigen eliciting potent antiviral control even at such late infection stage. PMID- 18408148 TI - Daily supplementation with iron increases lipid peroxidation in young women with low iron stores. AB - The aim of this study was to determine whether women with low iron stores (plasma ferritin 30% of known plant virus species, many of which are of great agricultural significance. These viruses have a positive sense RNA genome that is approximately 10 kb long and contains a single long ORF. The ORF is translated into a large polyprotein, which is cleaved into approximately 10 mature proteins. We report the discovery of a short ORF embedded within the P3 cistron of the polyprotein but translated in the +2 reading-frame. The ORF, termed pipo, is conserved and has a strong bioinformatic coding signature throughout the large and diverse Potyviridae family. Mutations that knock out expression of the PIPO protein in Turnip mosaic potyvirus but leave the polyprotein amino acid sequence unaltered are lethal to the virus. Immunoblotting with antisera raised against two nonoverlapping 14-aa antigens, derived from the PIPO amino acid sequence, reveals the expression of an approximately 25-kDa PIPO fusion product in planta. This is consistent with expression of PIPO as a P3-PIPO fusion product via ribosomal frameshifting or transcriptional slippage at a highly conserved G(1-2)A(6-7) motif at the 5' end of pipo. This discovery suggests that other short overlapping genes may remain hidden even in well studied virus genomes (as well as cellular organisms) and demonstrates the utility of the software package MLOGD as a tool for identifying such genes. PMID- 18408155 TI - The genome of Bacillus coahuilensis reveals adaptations essential for survival in the relic of an ancient marine environment. AB - The Cuatro Cienegas Basin (CCB) in the central part of the Chihuahan desert (Coahuila, Mexico) hosts a wide diversity of microorganisms contained within springs thought to be geomorphological relics of an ancient sea. A major question remaining to be answered is whether bacteria from CCB are ancient marine bacteria that adapted to an oligotrophic system poor in NaCl, rich in sulfates, and with extremely low phosphorus levels (<0.3 microM). Here, we report the complete genome sequence of Bacillus coahuilensis, a sporulating bacterium isolated from the water column of a desiccation lagoon in CCB. At 3.35 Megabases this is the smallest genome sequenced to date of a Bacillus species and provides insights into the origin, evolution, and adaptation of B. coahuilensis to the CCB environment. We propose that the size and complexity of the B. coahuilensis genome reflects the adaptation of an ancient marine bacterium to a novel environment, providing support to a "marine isolation origin hypothesis" that is consistent with the geology of CCB. This genomic adaptation includes the acquisition through horizontal gene transfer of genes involved in phosphorous utilization efficiency and adaptation to high-light environments. The B. coahuilensis genome sequence also revealed important ecological features of the bacterial community in CCB and offers opportunities for a unique glimpse of a microbe-dominated world last seen in the Precambrian. PMID- 18408157 TI - An essential sensor histidine kinase controlled by transmembrane helix interactions with its auxiliary proteins. AB - Two-component signal transduction systems with membrane-embedded sensor histidine kinases are believed to recognize environmental signals and transduce this information over the cellular membrane to influence the activity of a transcription factor to which they are mated. The YycG sensor kinase of Bacillus subtilis, containing two transmembrane helices, is subject to a complicated activity-control circuit involving two other proteins with N-terminal transmembrane helices, YycH and YycI. Truncation studies of YycH and YycI demonstrated that the individual transmembrane helices of these proteins are sufficient to adjust YycG activity, indicating that this control is achieved at the membrane level. A replica exchange molecular dynamics computational approach generated in silico structural models of the transmembrane helix complex that informed mutagenesis studies of the YycI transmembrane helix supporting the accuracy of the in silico model. The results predict that signal recognition by any of the extracellular domains of the sensor histidine kinase YycG or the associated proteins YycH and YycI is transmitted across the cellular membrane by subtle alterations in the positions of the helices within the transmembrane complex of the three proteins. PMID- 18408158 TI - Salt tolerance of Arabidopsis thaliana requires maturation of N-glycosylated proteins in the Golgi apparatus. AB - Protein N-glycosylation in the endoplasmic reticulum (ER) and in the Golgi apparatus is an essential process in eukaryotic cells. Although the N glycosylation pathway in the ER has been shown to regulate protein quality control, salt tolerance, and cellulose biosynthesis in plants, no biological roles have been linked functionally to N-glycan modifications that occur in the Golgi apparatus. Herein, we provide evidence that mutants defective in N-glycan maturation, such as complex glycan 1 (cgl1), are more salt-sensitive than wild type. Salt stress caused growth inhibition, aberrant root-tip morphology, and callose accumulation in cgl1, which were also observed in an ER oligosaccharyltransferase mutant, staurosporin and temperature sensitive 3a (stt3a). Unlike stt3a, cgl1 did not cause constitutive activation of the unfolded protein response. Instead, aberrant modification of the plasma membrane glycoprotein KORRIGAN 1/RADIALLY SWOLLEN 2 (KOR1/RSW2) that is necessary for cellulose biosynthesis occurred in cgl1 and stt3a. Genetic analyses identified specific interactions among rsw2, stt3a, and cgl1 mutations, indicating that the function of KOR1/RSW2 protein depends on complex N-glycans. Furthermore, cellulose deficient rsw1-1 and rsw2-1 plants were also salt-sensitive. These results establish that plant protein N-glycosylation functions beyond protein folding in the ER and is necessary for sufficient cell-wall formation under salt stress. PMID- 18408159 TI - A relaxed active site after exon ligation by the group I intron. AB - During RNA maturation, the group I intron promotes two sequential phosphorotransfer reactions resulting in exon ligation and intron release. Here, we report the crystal structure of the intron in complex with spliced exons and two additional structures that examine the role of active-site metal ions during the second step of RNA splicing. These structures reveal a relaxed active site, in which direct metal coordination by the exons is lost after ligation, while other tertiary interactions are retained between the exon and the intron. Consistent with these structural observations, kinetic and thermodynamic measurements show that the scissile phosphate makes direct contact with metals in the ground state before exon ligation and in the transition state, but not after exon ligation. Despite no direct exonic interactions and even in the absence of the scissile phosphate, two metal ions remain bound within the active site. Together, these data suggest that release of the ligated exons from the intron is preceded by a change in substrate-metal coordination before tertiary hydrogen bonding contacts to the exons are broken. PMID- 18408160 TI - Nonuniform distribution of glucosinolates in Arabidopsis thaliana leaves has important consequences for plant defense. AB - The spatial distribution of plant defenses within a leaf may be critical in explaining patterns of herbivory. The generalist lepidopteran larvae, Helicoverpa armigera (the cotton bollworm), avoided the midvein and periphery of Arabidopsis thaliana rosette leaves and fed almost exclusively on the inner lamina. This feeding pattern was attributed to glucosinolates because it was not evident in a myrosinase mutant that lacks the ability to activate glucosinolate defenses by hydrolysis. To measure the spatial distribution of glucosinolates in A. thaliana leaves at a fine scale, we constructed ion intensity maps from MALDI-TOF (matrix assisted laser desorption/ionization-time of flight) mass spectra. The major glucosinolates were found to be more abundant in tissues of the midvein and the periphery of the leaf than the inner lamina, patterns that were validated by HPLC analyses of dissected leaves. In addition, there were differences in the proportions of the three major glucosinolates in different leaf regions. Hence, the distribution of glucosinolates within the leaf appears to control the feeding preference of H. armigera larvae. The preferential allocation of glucosinolates to the periphery may play a key role in the defense of leaves by creating a barrier to the feeding of chewing herbivores that frequently approach leaves from the edge. PMID- 18408161 TI - Linking functionally related genes by sensitive and quantitative characterization of genetic interaction profiles. AB - Describing at a genomic scale how mutations in different genes influence one another is essential to the understanding of how genotype correlates with phenotype and remains a major challenge in biology. Previous studies pointed out the need for accurate measurements of not only synthetic but also buffering interactions in the characterization of genetic networks and functional modules. We developed a sensitive and efficient method that allows such measurements at a genomic scale in yeast. In a pilot experiment (41 genome-wide screens), we quantified the fitness of 140,000 double deletion strains relative to the corresponding single mutants and identified many genetic interactions. In addition to synthetic growth defects (validated experimentally with factors newly identified as genetically interfering with mRNA degradation), most of the identified genetic interactions measured weak epistatic effects. These weak effects, rarely meaningful when considered individually, were crucial to defining specific signatures for many gene deletions and had a major contribution in defining clusters of functionally related genes. PMID- 18408162 TI - Proton order in the ice crystal surface. AB - The physics of the ice crystal surface and its interaction with adsorbates are not only of fundamental interest but also of considerable importance to terrestrial and planetary chemistry. Yet the atomic-level structure of even the pristine ice surface at low temperature is still far from well understood. This computational study focuses on the pattern of dangling H and dangling O (lone pairs) atoms at the basal ice surface. Dangling atoms serve as binding sites for adsorbates capable of hydrogen- and electrostatic bonding. Extension of the well known orientational disorder ("proton disorder") of bulk crystal ice to the surface would naturally suggest a disordered dangling atom pattern; however, extensive computer simulations employing two different empirical potentials indicate significant free energy preference for a striped phase with alternating rows of dangling H and dangling O atoms, as suggested long ago by Fletcher [Fletcher NH (1992) Philos Mag 66:109-115]. The presence of striped phase domains within the basal surface is consistent with the hitherto unexplained minor fractional peaks in the helium diffraction pattern observed 10 years ago. Compared with the disordered model, the striped model yields improved agreement between computations and experimental ppp-polarized sum frequency generation spectra. PMID- 18408163 TI - Cassandra retrotransposons carry independently transcribed 5S RNA. AB - We report a group of TRIMs (terminal-repeat retrotransposons in miniature), which are small nonautonomous retrotransposons. These elements, named Cassandra, universally carry conserved 5S RNA sequences and associated RNA polymerase (pol) III promoters and terminators in their long terminal repeats (LTRs). They were found in all vascular plants investigated. Uniquely for LTR retrotransposons, Cassandra produces noncapped, polyadenylated transcripts from the 5S pol III promoter. Capped, read-through transcripts containing Cassandra sequences can also be detected in RNA and in EST databases. The predicted Cassandra RNA 5S secondary structures resemble those for cellular 5S rRNA, with high information content specifically in the pol III promoter region. Genic integration sites are common for Cassandra, an unusual feature for abundant retrotransposons. The 5S in each LTR produces a tandem 5S arrangement with an inter-5S spacing resembling that of cellular 5S. The distribution of 5S genes is very variable in flowering plants and may be partially explained by Cassandra activity. Cassandra thus appears both to have adapted a ubiquitous cellular gene for ribosomal RNA for use as a promoter and to parasitize an as-yet-unidentified group of retrotransposons for the proteins needed in its lifecycle. PMID- 18408164 TI - Membrane damage by human islet amyloid polypeptide through fibril growth at the membrane. AB - Fibrillar protein deposits (amyloid) in the pancreatic islets of Langerhans are thought to be involved in death of the insulin-producing islet beta cells in type 2 diabetes mellitus. It has been suggested that the mechanism of this beta cell death involves membrane disruption by human islet amyloid polypeptide (hIAPP), the major constituent of islet amyloid. However, the molecular mechanism of hIAPP induced membrane disruption is not known. Here, we propose a hypothesis that growth of hIAPP fibrils at the membrane causes membrane damage. We studied the kinetics of hIAPP-induced membrane damage in relation to hIAPP fibril growth and found that the kinetic profile of hIAPP-induced membrane damage is characterized by a lag phase and a sigmoidal transition, which matches the kinetic profile of hIAPP fibril growth. The observation that seeding accelerates membrane damage supports the hypothesis. In addition, variables that are well known to affect hIAPP fibril formation, i.e., the presence of a fibril formation inhibitor, hIAPP concentration, and lipid composition, were found to have the same effect on hIAPP induced membrane damage. Furthermore, electron microscopy analysis showed that hIAPP fibrils line the surface of distorted phospholipid vesicles, in agreement with the notion that hIAPP fibril growth at the membrane and membrane damage are physically connected. Together, these observations point toward a mechanism in which growth of hIAPP fibrils, rather than a particular hIAPP species, is responsible for the observed membrane damage. This hypothesis provides an additional mechanism next to the previously proposed role of oligomers as the main cytotoxic species of amyloidogenic proteins. PMID- 18408165 TI - Role of the familial Dutch mutation E22Q in the folding and aggregation of the 15 28 fragment of the Alzheimer amyloid-beta protein. AB - Amyloid fibrils, large ordered aggregates of amyloid beta proteins (Abeta), are clinical hallmarks of Alzheimer's disease (AD). The aggregation properties of amyloid beta proteins can be strongly affected by single-point mutations at positions 22 and 23. The Dutch mutation involves a substitution at position 22 (E22Q) and leads to increased deposition rates of the AbetaE22Q peptide onto preseeded fibrils. We investigate the effect of the E22Q mutation on two key regions involved in the folding and aggregation of the Abeta peptide through replica exchange molecular dynamics simulations of the 15-28 fragment of the Abeta peptide. The Abeta15-28 peptide encompasses the 22-28 region that constitutes the most structured part of the Abeta peptide (the E22-K28 bend), as well as the central hydrophobic cluster (CHC) (segment 17-21), the primary docking site for Abeta monomers depositing onto fibrils. Our simulations show that the 22-28 bend is preserved in the Abeta(15-28) peptide and that the CHC, which is mostly unstructured, interacts with this bend region. The E22Q mutation does not affect the structure of the bend but weakens the interactions between the CHC and the bend. This leads to an increased population of beta-structure in the CHC. Our analysis of the fibril elongation reaction reveals that the CHC adopts a beta-strand conformation in the transition state ensemble, and that the E22Q mutation increases aggregation rates by lowering the barrier for Abeta monomer deposition onto a fibril. Thermodynamic signatures of this enhanced fibrillization process from our simulations are in good agreement with experimental observations. PMID- 18408166 TI - Simulation of Top7-CFr: a transient helix extension guides folding. AB - Protein structures often feature beta-sheets in which adjacent beta-strands have large sequence separation. How the folding process orchestrates the formation and correct arrangement of these strands is not comprehensively understood. Particularly challenging are proteins in which beta-strands at the N and C termini are neighbors in a beta-sheet. The N-terminal beta-strand is synthesized early on, but it can not bind to the C terminus before the chain is fully synthesized. During this time, there is a danger that the beta-strand at the N terminus interacts with nearby molecules, leading to potentially harmful aggregates of incompletely folded proteins. Simulations of the C-terminal fragment of Top7 show that this risk of misfolding and aggregation can be avoided by a "caching" mechanism that relies on the "chameleon" behavior of certain segments. PMID- 18408168 TI - Comparison of SCImago journal rank indicator with journal impact factor. AB - The application of currently available sophisticated algorithms of citation analysis allows for the incorporation of the "quality" of citations in the evaluation of scientific journals. We sought to compare the newly introduced SCImago journal rank (SJR) indicator with the journal impact factor (IF). We retrieved relevant information from the official Web sites hosting the above indices and their source databases. The SJR indicator is an open-access resource, while the journal IF requires paid subscription. The SJR indicator (based on Scopus data) lists considerably more journal titles published in a wider variety of countries and languages, than the journal IF (based on Web of Science data). Both indices divide citations to a journal by articles of the journal, during a specific time period. However, contrary to the journal IF, the SJR indicator attributes different weight to citations depending on the "prestige" of the citing journal without the influence of journal self-citations; prestige is estimated with the application of the PageRank algorithm in the network of journals. In addition, the SJR indicator includes the total number of documents of a journal in the denominator of the relevant calculation, whereas the journal IF includes only "citable" articles (mainly original articles and reviews). A 3 yr period is analyzed in both indices but with the use of different approaches. Regarding the top 100 journals in the 2006 journal IF ranking order, the median absolute change in their ranking position with the use of the SJR indicator is 32 (1st quartile: 12; 3rd quartile: 75). Although further validation is warranted, the novel SJR indicator poses as a serious alternative to the well-established journal IF, mainly due to its open-access nature, larger source database, and assessment of the quality of citations. PMID- 18408169 TI - Stem cell researchers find their niche. AB - The EuroSTELLS Workshop ;Stem Cell Niches', organised by Anna Bigas, Ernest Arenas and Pasqualino Loi, took place in January 2008 in Barcelona, Spain. The goal of the conference was to promote scientific collaboration and synergy between stem cell researchers worldwide and those in the EuroSTELLS consortia (an initiative of the European Science Foundation EUROCORES Programme), and to stimulate discussion of the latest results in the field of stem cell niches. PMID- 18408167 TI - An essential role for SRC-activated STAT-3 in 14,15-EET-induced VEGF expression and angiogenesis. AB - To understand the molecular mechanisms underlying 14,15-epoxyeicosatrienoic acid (14,15-EET)-induced angiogenesis, here we have studied the role of signal transducer and activator of transcription-3 (STAT-3). 14,15-EET stimulated the tyrosine phosphorylation of STAT-3 and its translocation from the cytoplasm to the nucleus in human dermal microvascular endothelial cells (HDMVECs). Adenovirus mediated delivery of dominant negative STAT-3 substantially inhibited 14,15-EET induced HDMVEC migration, and tube formation and Matrigel plug angiogenesis. 14,15-EET activated Src, as measured by its tyrosine phosphorylation and blockade of its activation by adenovirus-mediated expression of its dominant negative mutant, significantly attenuated 14,15-EET-induced STAT-3 phosphorylation in HDMVECs and the migration and tube formation of these cells and Matrigel plug angiogenesis. 14,15-EET induced the expression of vascular endothelial cell growth factor (VEGF) in a time- and Src-STAT-3-dependent manner in HDMVECs. Transfac analysis of VEGF promoter revealed the presence of STAT-binding elements and 14,15-EET induced STAT-3 binding to this promoter in vivo, and this interaction was inhibited by suppression of Src-STAT-3 signaling. Neutralizing anti-VEGF antibodies completely blocked 14,15-EET-induced HDMVEC migration and tube formation and Matrigel plug angiogenesis. These results reveal that Src dependent STAT-3-mediated VEGF expression is a major mechanism of 14,15-EET induced angiogenesis. PMID- 18408170 TI - Intimate partner violence among pregnant Thai women. AB - The purposes of this study are to describe intimate partner violence (IPV) among pregnant women in Thailand and compare maternal characteristics, health practices during pregnancy, and maternal health between abused and nonabused pregnant women. Of 475 pregnant Thai women, 13.1% report ever being abused, whereas 4.8% report physical abuse during pregnancy. Women abused during pregnancy, compared to nonabused women, are more likely to be younger, unmarried, have low income, be unemployed, and report that the pregnancy was unwanted. They also report lower levels of positive health practices and higher levels of depressive symptoms. Results demonstrate a need for screening of IPV among pregnant Thai women. PMID- 18408171 TI - Prevalence and patterns of sexual assault across the life span among incarcerated women. AB - This study investigated the prevalence of and associations among sexual assault by life stage (childhood, adolescence, or adulthood) and perpetrator (family, stranger, friend, or partner) via a survey of a statewide sample of incarcerated women (N = 484). Participants were 18 to 56 years old, and the majority were White (56%). Results demonstrate higher rates of sexual assault in childhood (35%) and adulthood (22%) as compared with adolescence (14%). Logistic regression analyses revealed significant associations between childhood sexual assault by family and adulthood sexual assault by friend, stranger, and partner; adolescent sexual assault was not significantly associated with sexual assault in childhood or adulthood. These findings suggest that the lifetime sexual victimization pattern of incarcerated women differs from that seen in the general population. PMID- 18408172 TI - Sexual boundaries: an examination of the importance of talking before touching. AB - This study examined a woman's clearly articulated sexual boundary and its effect on college students' discrimination of when a woman wants her date to stop making sexual advances. Male and female participants listened to an audio recording of a date rape vignette and signaled when the man should stop making sexual advances. Relative to participants in the no-boundary condition, participants who heard a discussion including a sexual boundary before intimate physical contact occurred displayed significantly shorter latencies to identify the inappropriateness of the man's behavior. No significant difference was observed between male and female participants. PMID- 18408173 TI - Development of the scale of economic abuse. AB - Economic abuse is part of the pattern of behaviors used by batterers to maintain power and control over their partners. However, no measure of economic abuse exists. This study describes the development of the Scale of Economic Abuse, which was designed to fill this gap. Interviews were conducted with 103 survivors of domestic abuse, each of whom responded to measures of economic, physical, and psychological abuse as well as economic hardship. Results provide evidence for the reliability and validity of the scale. This study is an important first step toward understanding the extent and impact of economic abuse experienced by survivors. PMID- 18408174 TI - The men's domestic abuse check-up: a protocol for reaching the nonadjudicated and untreated man who batters and who abuses substances. AB - Batterer intervention programs primarily work with individuals mandated to participate. Commonly, attrition is high and outcomes are modest. Motivational enhancement therapy (MET), most widely studied in the substance abuse field, offers a potentially effective approach to improving self-referral to treatment, program retention, treatment compliance, and posttreatment outcomes among men who batter and who abuse substances. A strategy for using a catalyst variant of MET (a "check-up") to reach untreated, nonadjudicated perpetrators is described in detail. Unique challenges in evaluating the success of this approach are discussed, including attending to victim safety and determining indicators of increased motivation for change. PMID- 18408175 TI - Prospective epidemiological cohort study of reduced risk for incident cataract with vigorous physical activity and cardiorespiratory fitness during a 7-year follow-up. AB - PURPOSE: To test whether greater vigorous physical activity (kilometers per week run) and greater cardiorespiratory fitness (10-km race performance in meters per second) reduce the incidence of clinically diagnosed cataract. METHODS: Prospective cohort study of self-reported clinical diagnosis of cataract in nondiabetic, nonvegetarian, and nonsmoking male (n = 29,025) and female runners (n = 11,967). RESULTS: Incident cataracts were reported by 733 (2.53%) men and 179 (1.50%) women during (mean +/- SE) 7.74 +/- 0.01 and 7.42 +/- 0.02 years of follow-up, respectively. The risk for incident cataract increased with BMI, such that the risk in men > 27.5 kg/m(2) was 88% larger than in men < 20 kg/m(2). Men's cataract risk declined significantly in relation to running distance (P = 0.01), even when adjusted for BMI. Men who ran > or = 64 km/wk had 35% lower risk for cataract than those reporting < 16 km/wk (28% lower risk when adjusted for BMI). In addition, men with greater cardiorespiratory fitness were at significantly less risk for development of cataract than were the least fit men. This result was not accounted for by adjustment for running distance or BMI. Compared with the least fit men, those who ran faster than 4.75 m/s had 50% lower risk for incident cataract (43% lower when adjusted for km/wk and BMI). CONCLUSIONS: These data suggest that the men's cataract risk decreased in association with lower BMI, greater physical activity, and greater cardiorespiratory fitness, the latter being statistically independent of both BMI and physical activity. The study limitations include the absence of confirmation of the clinical diagnosis and the lack of specificity of the type of cataract. PMID- 18408176 TI - Quantitative subanalysis of optical coherence tomography after treatment with ranibizumab for neovascular age-related macular degeneration. AB - PURPOSE: To investigate the effects of ranibizumab on retinal morphology in patients with neovascular age-related macular degeneration (AMD) using optical coherence tomography (OCT) quantitative subanalysis. METHODS: Data from 95 patients receiving intravitreal ranibizumab for neovascular AMD were collected. StratusOCT images were analyzed using custom software that allows precise positioning of prespecified boundaries on every B-scan. Changes in thickness/volume of the retina, subretinal fluid (SRF), subretinal tissue (SRT), and pigment epithelial detachments (PEDs) at week 1 and at months 1, 3, 6, and 9 after treatment were calculated. RESULTS: Total retinal volume reached its nadir at month 1, with an average reduction of 0.43 mm(3) (P < 0.001). By month 9, this initial change had been reduced to a mean reduction of 0.32 mm(3) (P = 0.0011). Total SRF volume reached its lowest level by month 1, with an average reduction of 0.24 mm(3) (P < 0.001). This reduction lessened subsequently, to 0.18 mm(3), by month 9. There was an average 0.3-mm(3) decrease in total PED volume by month 1 (P < 0.001), and this later declined further, to 0.45 mm(3), by month 9 (P = 0.0014). Total SRT volume was reduced by an average of 0.07 mm(3) at month 1 (P = 0.0159) and subsequently remained constant. CONCLUSIONS: Although neurosensory retinal edema and SRF showed an early reduction to nadir after the initiation of ranibizumab therapy, the effect on the retina was attenuated over time, suggesting possible tachyphylaxis. PED volume showed a slower but progressive reduction. Manual quantitative OCT subanalysis may allow a more precise understanding of anatomic outcomes and their correlation with visual acuity. PMID- 18408177 TI - Computerized analysis of retinal vessel width and tortuosity in premature infants. AB - PURPOSE: To determine, with novel software, the feasibility of measuring the tortuosity and width of retinal veins and arteries from digital retinal images of infants at risk of retinopathy of prematurity (ROP). METHODS: The Computer-Aided Image Analysis of the Retina (CAIAR) program was developed to enable semiautomatic detection of retinal vasculature and measurement of vessel tortuosity and width from digital images. CAIAR was tested for accuracy and reproducibility of tortuosity and width measurements by using computer-generated vessel-like lines of known frequency, amplitude, and width. CAIAR was then tested by using clinical digital retinal images for correlation of vessel tortuosity and width readings compared with expert ophthalmologist grading. RESULTS: When applied to 16 computer-generated sinusoidal vessels, the tortuosity measured by CAIAR correlated very well with the known values. Width measures also increased as expected. When the CAIAR readings were compared with five expert ophthalmologists' grading of 75 vessels on 10 retinal images, moderate correlation was found in 10 of the 14 tortuosity output calculations (Spearman rho = 0.618-0.673). Width was less well correlated (rho = 0.415). CONCLUSIONS: The measures of tortuosity and width in CAIAR were validated using sequential model vessel analysis. On comparison of CAIAR output with assessments made by expert ophthalmologists, CAIAR correlates moderately with tortuosity grades, but less well with width grades. CAIAR offers the opportunity to develop an automated image analysis system for detecting the vascular changes at the posterior pole, which are becoming increasingly important in diagnosing treatable ROP. PMID- 18408179 TI - Size overlap between benign melanocytic choroidal nevi and choroidal malignant melanomas. AB - PURPOSE: To estimate size overlap between large choroidal nevi and small choroidal melanomas by using plotted frequency distributions of tumor size. METHODS: Frequency distributions of largest linear basal diameter (LBD) and thickness (TH) of choroidal nevi and melanomas were plotted from published data and cases in the senior author's practice. Relative frequencies of choroidal nevi and melanomas were estimated from published data. Relative frequency distributions of the tumors were plotted to illustrate the extent of overlap between them. RESULTS: Comparison of plotted frequency distribution curves for thickness indicated that there were approximately 125 nevi for every melanoma in the TH range 1.5 to 2 mm, approximately 25 nevi for every melanoma in the TH range 2 to 2.5 mm, and approximately 5 nevi for every melanoma in the TH range 2.5 to 3 mm. Similarly, comparison of the plotted frequency distribution curves for LBD of these tumor types indicated that there were approximately 70 nevi for every choroidal melanoma in the LBD range 5 to 6 mm, approximately 10 nevi for every melanoma in the LBD range 6 to 7 mm, and approximately 3 nevi for every melanoma in the LBD range 7 to 8 mm. CONCLUSIONS: Because of the markedly greater cumulative lifetime incidence of choroidal nevi, the results of this analysis suggest considerable size overlap between larger nevi and smaller melanomas. Attempts to classify small melanocytic choroidal tumors clinically as benign nevi versus malignant melanomas on the basis of tumor size appear likely to result in multiple misclassifications. PMID- 18408178 TI - The proteasome: a target of oxidative damage in cultured human retina pigment epithelial cells. AB - PURPOSE: Dysfunction of the ubiquitin-proteasome pathway (UPP) is associated with several age-related degenerative diseases. The objective of this study was to investigate the effect of oxidative stress on the UPP in cultured human retina pigment epithelial cells. METHODS: To mimic physiological oxidative stress, ARPE 19 cells were exposed to continuously generated H2O(2) or A2E-mediated photooxidation. Proteasome activity was monitored using fluorogenic peptides as substrates. The ubiquitin conjugation activity and activities of E1 and E2 were determined by the thiolester assays. Levels of ubiquitin and ubiquitin conjugates were determined by Western blotting. RESULTS: Exposure of ARPE-19 cells to 40 to 50 microM H2O(2) for 4 hours resulted in a 30% to 50% reduction in all three peptidase activities of the proteasome. Similarly, exposure of A2E-loaded ARPE-19 cells to blue light resulted in a 40% to 60% reduction in proteasome activity. Loading of A2E or exposure to blue light alone had little effect on proteasome activity. In contrast, exposure of ARPE-19 to low levels of H2O(2) (10 microM) stimulated ubiquitin conjugation activity. Loading of A2E, with or without exposure to blue light, upregulated the levels of ubiquitin-activating enzyme and increased conjugation activity. Exposure to H2O(2) or A2E-mediated photooxidation also resulted in a twofold to threefold increase in levels of endogenous ubiquitin conjugates. CONCLUSIONS: These data show that the proteasome in ARPE-19 is susceptible to oxidative inactivation, whereas activities of the ubiquitin conjugating enzymes are more resistant to oxidative stress. Oxidative inactivation of the proteasome appears to be one of the mechanisms underlying stress-induced accumulation of ubiquitin conjugates in the cells. PMID- 18408181 TI - Anterior chamber angle measurement with anterior segment optical coherence tomography: a comparison between slit lamp OCT and Visante OCT. AB - PURPOSE: To compare anterior chamber angle measurements obtained from two anterior segment optical coherence tomography (OCT) instruments and to evaluate their agreements and interobserver reproducibility. METHODS: Forty-nine eyes from 49 healthy normal subjects were studied. The anterior chamber angle was imaged with the Visante anterior segment OCT (Carl Zeiss Meditec, Dublin, CA) and the slit lamp OCT (SLOCT, Heidelberg Engineering, GmbH, Dossenheim, Germany) on one randomly selected eye in each subject and measured by two independent observers. The angle-opening distance (AOD 500), the trabecular-iris angle (TIA 500), and the trabecular-iris space area (TISA 500) at the nasal and temporal angles were measured. The agreements between SLOCT and Visante OCT measurements and the interobserver reproducibility were evaluated. RESULTS: The mean nasal/temporal anterior chamber angles measured by Visante OCT and SLOCT were 527 +/- 249/572 +/ 275 microm (AOD), 0.180 +/- 0.091/0.193 +/- 0.102 mm(2) (TISA), and 38.1 +/- 12.3/39.6 +/- 13.2 degrees (TIA); and 534 +/- 234/628 +/- 254 microm (AOD), 0.191 +/- 0.089/0.217 +/- 0.093 mm(2)(TISA), and 37.8 +/- 10.1/40.6 +/- 10.7 degrees (TIA), respectively. No significant difference was found between Visante OCT and SLOCT measurements except the temporal TISA (P = 0.034). The interobserver coefficient of variation ranged between 4.4% and 7.8% for Visante OCT and 4.9% and 7.0% for SLOCT. The spans of 95% limits of agreement of the nasal/temporal angle measurements between Visante OCT and SLOCT were 437/531 mm(2), 0.174/0.186 mm(2), and 25.3/28.0 degrees for AOD, TISA, and TIA, respectively. CONCLUSIONS: Although Visante OCT and SLOCT demonstrate high interobserver reproducibility for anterior chamber angle measurements, their agreement was poor. PMID- 18408180 TI - The Leber congenital amaurosis protein AIPL1 functions as part of a chaperone heterocomplex. AB - PURPOSE: AIPL1 mutations cause the severe inherited blindness Leber congenital amaurosis (LCA). The similarity of AIPL1 to tetratricopeptide repeat (TPR) cochaperones that interact with the chaperone Hsp90 and the ability of AIPL1 to suppress the aggregation of NUB1 fragments in a chaperone-like manner suggest that AIPL1 might function as part of a chaperone heterocomplex facilitating retinal protein maturation. In this study the interaction of AIPL1 with molecular chaperones is revealed and functionally characterized. METHODS: AIPL1-interacting proteins were identified using a yeast two-hybrid system, and the effect of AIPL1 pathogenic mutations and sequence requirements mediating the identified interactions were investigated. The interactions were validated by a comprehensive set of biochemical assays, and the ability of the AIPL1-binding partners to cooperate with AIPL1 in the suppression of NUB1 fragment aggregation was assessed. RESULTS: AIPL1 interacts with the molecular chaperones Hsp90 and Hsp70. Mutations within the TPR domain of AIPL1 or removal of the chaperone TPR acceptor site abolished the interactions. Importantly, LCA-causing mutations in AIPL1 also compromised these interactions, suggesting that the essential function of AIPL1 in photoreceptors may involve the interaction with Hsp90 and Hsp70. Examination of the role of these chaperones in AIPL1 chaperone activity demonstrated that AIPL1 cooperated with Hsp70, but not with Hsp90, to suppress the formation of NUB1 inclusions. CONCLUSIONS: These findings suggest that AIPL1 may cooperate with both Hsp70 and Hsp90 within a retina-specific chaperone heterocomplex and that the specialized role of AIPL1 in photoreceptors may therefore be facilitated by these molecular chaperones. PMID- 18408182 TI - Focal macular electroretinograms in eyes with wet-type age-related macular degeneration. AB - PURPOSE: To study the properties of the focal macular electroretinograms (fmERGs) in eyes with untreated wet-type age-related macular degeneration (AMD). METHODS: fmERGs were recorded from 157 eyes of 157 consecutive patients with untreated wet type AMD (113 men, 44 women; age, 71.3 +/- 8.0 years). The fmERGs were recorded under direct fundus observation using a modified infrared fundus camera and a 15 degrees stimulus spot. Amplitudes and implicit times of the fmERGs recorded from the AMD patients were compared with those from 21 age-similar healthy controls. RESULTS: The amplitudes of fmERGs in the AMD patients were significantly smaller (P < 0.001) and the implicit times were significantly longer (P < 0.001) than the corresponding values in the control eyes. There was a significant correlation between amplitude and implicit time of the fmERG and visual acuity (logMAR), but the degree of correlation was weak. The difference in the b/a amplitude ratio between the AMD patients and healthy controls was not significant. CONCLUSIONS: The significant reduction in amplitude and the severe delay in implicit times of a- and b-waves of the fmERGs indicated significant functional alterations in the inner and the outer retinal layers of the macular area of eyes with wet-type AMD. PMID- 18408183 TI - Human Muller stem cell (MIO-M1) transplantation in a rat model of glaucoma: survival, differentiation, and integration. AB - PURPOSE: Stem cell transplantation is a potential treatment strategy for neurodegenerative diseases such as glaucoma. The Muller stem cell line MIO-M1 can be differentiated to produce retinal neurons and glia. The survival, migration, differentiation, and integration of MIO-M1 cells were investigated in a rat model of glaucoma. The effect of modulating the retinal environment with either chondroitinase ABC or erythropoietin was also studied. METHODS: Intraocular pressure was chronically increased unilaterally by using a laser glaucoma model in adult rats. EGFP-transduced MIO-M1 cells were transplanted into the vitreous or subretinal space of glaucomatous or untreated eyes. Oral immune suppressants were administered to reduce xenograft rejection. Survival, migration, differentiation, and integration of grafted cells were assessed by immunohistochemistry. RESULTS: Transplanted cells survived for 2 to 3 weeks in vivo, although microglia/macrophage infiltration and a reduction in graft survival were seen by 4 weeks. Grafted cells displayed a migratory phenotype with an elongated bipolar shape often oriented toward the retina. Transplanted cells expressed markers such as PSA-NCAM, GFAP, and beta-III-tubulin. The host retina was resistant to MIO-M1 migration, but modification of the local environment with erythropoietin or chondroitinase ABC facilitated retinal infiltration by MIO-M1 cells. CONCLUSIONS: The results demonstrate that differentiating MIO-M1 cells within the glaucomatous eye produced cells that expressed neuronal and glial cell markers. The retina was relatively resistant to transplant integration, and long term xenograft survival was limited. However, local modulation of the retinal environment enhanced the integration of MIO-M1 cells into the glaucomatous retina. PMID- 18408184 TI - Dendritic cells augment choroidal neovascularization. AB - PURPOSE: Dendritic cells (DCs) are innate immune cells that have recently been shown to support angiogenesis in tumors, endometriosis, and lymph nodes. A major cause of legal blindness is wet age-related macular degeneration (wet ARMD), wherein abnormal blood vessels grow under the retina, an abnormality also referred to as choroidal neovascularization (CNV). The purpose of the present study was to investigate the role of DCs in the development of CNV. METHODS: Laser photocoagulation was used to induce CNV in C57BL/6J mice. The authors analyzed CNV lesions for the presence of DCs using flow cytometry and immunostaining at designated times. They also analyzed the effects of intravenous DC transplantation on CNV development by measuring the lesion area using confocal microscopy 1 week after laser injury. RESULTS: The authors analyzed CNV lesions for the presence of DCs by flow cytometry and observed that CD11c(+) major histocompatibility complex (MHC) class II(+) DCs transiently infiltrated the CNV lesions, reaching a peak at 2 to 4 days after laser injury. These DCs were mostly immature (CD11c(+) MHCII(low)) and expressed vascular endothelial growth factor receptor 2. Immunostaining of laser-induced CNV lesions confirmed that DCs are located at the sites of newly formed blood vessels. Intravenously injected DCs incorporated into the CNV lesions. However, only immature DCs enhanced CNV size. CONCLUSIONS: These results suggest a role for DCs in promoting angiogenesis and lesion growth in laser-induced CNV. The present data suggest that DCs may represent potential cellular targets for therapeutic intervention in wet ARMD. PMID- 18408185 TI - Innervation of tissue-engineered recombinant human collagen-based corneal substitutes: a comparative in vivo confocal microscopy study. AB - PURPOSE: To compare reinnervation in recombinant human collagen-based corneal substitutes with allografts during a 1-year postimplantation follow-up period in pigs. A retrospective comparison to innervation in porcine collagen-based biosynthetic grafts was also performed. METHODS: Pigs received a corneal allograft or a substitute made of either recombinant human type-I or -III collagen. In vivo confocal microscopic examination of the central cornea of surgical and untouched control eyes before surgery and at 2, 6, and 12 months after surgery was performed to quantify the number, density, and diameter of nerves at various corneal depths. RESULTS: By 12 months after surgery, the number and density of regenerated nerves in the anterior and deep anterior corneal stroma recovered to preoperative and control levels in both types of substitute grafts and in the allografts. In the subepithelial and subbasal regions, however, significantly fewer nerves were detected relative to those in control subjects at 12 months, regardless of graft type (P < 0.05), similar to the behavior of porcine collagen-based biosynthetic grafts. An absence of thick stromal nerve trunks (diameter, >10 mum) in all grafts, irrespective of material type, indicated that nerve regeneration in grafts was accompanied by persistent morphologic changes. CONCLUSIONS: Nerve regeneration in recombinant human collagen-based biosynthetic corneal grafts proceeded similarly to that in allograft tissue, demonstrating the suitability of recombinant human collagen constructs as nerve-friendly corneal substitutes. Furthermore, only minor differences were noted between type-I and -III collagen grafts, indicating an insensitivity of nerve regeneration to initial collagen type. PMID- 18408186 TI - Development and characterization of an adult retinal explant organotypic tissue culture system as an in vitro intraocular stem cell transplantation model. AB - PURPOSE: To develop and characterize a retinal explant culture system to facilitate investigation of novel methods of improving retinal stem cell therapy. METHODS: Retinas explanted from adult rats were cultured in serum-free medium (B27/N2) or medium containing normal horse serum (NHS). Tissue viability was assessed by gross morphology, propidium iodide (PI) uptake, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. Muller progenitor cells (hMIO-M1), or mesenchymal stem cells (MSC) were placed on explants, to model intravitreal cell transplantation. Explants were compared with whole eyes, with or without experimental glaucoma and/or intravitreal cell transplantation. RESULTS: Explants cultured in B27/N2 medium were viable for at least 17 days, as assessed by the aforementioned parameters. NHS medium was associated with obvious tissue degradation, greater/more diffuse PI uptake, significant cell loss over time, and temporal increase in activated caspase-3(+) cells. Explants in B27/N2 medium strongly expressed beta-III-tubulin, neurofilament, NeuN, Brn3a, Thy-1, GFAP, vimentin, nestin, and glutamine synthetase, whereas immunoreactivity was weak in NHS medium and decreased further with time. Seven and 14 days after coculture or transplantation, glial reactivity (GFAP/vimentin expression) was highly upregulated in explants and eyes, respectively. Some grafted cells migrated into the retina, but most remained outside the inner limiting membrane. CONCLUSIONS: Retinal explants prepared using the described techniques and cultured in B27/N2 medium are viable for at least 2 weeks and mimic in vivo glial reactivity to transplantation while allowing few grafted cells to integrate. This system may be a useful in vitro model for investigating methods of enhancing retinal stem cell therapy. PMID- 18408187 TI - Abnormal vessel formation in the choroid of mice lacking tissue inhibitor of metalloprotease-3. AB - PURPOSE: Tissue inhibitor of metalloprotease (TIMP)-3 is an inhibitor of matrix metalloprotease (MMP) and regulates angiogenesis. In the eye, TIMP3 is tightly associated with Bruch's membrane. In this study, the authors analyzed mice lacking TIMP3 for retinal abnormalities. METHODS: Mice with targeted disruption of the Timp3 gene were generated (Timp3(-/-)) and bred into C57/Bl6 and CD1 backgrounds. Eyes were analyzed by light and electron microscopy. Vasculature was examined by scanning laser ophthalmoscopy, corrosion casts, and whole mount preparations. MMP activity was assessed by in situ zymography, angiogenic potential was evaluated by tube formation, and aortic ring assays and signaling pathways were studied by immunoblotting. RESULTS: TIMP3-deficient mice develop abnormal vessels with dilated capillaries throughout the choroid. Enhanced MMP activity in the choroid region of Timp3(-/-) eyes was detected when compared with controls. Timp3(-/-)-derived tissue showed an increased angiogenic activity over wild-type, an effect that could specifically be inhibited by recombinant TIMP3. Moreover, the antiangiogenic property of TIMP3 was demonstrated to reside within the C-terminal domain. When VEGFR2 inhibitor was added to Timp3(-/-) aortic explants, endothelial sprout formation was markedly reduced, which provided evidence for an unbalanced VEGF-mediated angiogenesis in Timp3(-/-) animals. Finally, angiogenic signaling pathways are activated in Timp3(-/-)-derived cells. CONCLUSIONS: These findings suggest that the distinct choroidal phenotype in mice lacking TIMP3 may be the result of a local disruption of extracellular matrix and angiogenic homeostasis, and they support an important role of TIMP3 in the regulation of choroidal vascularization. PMID- 18408189 TI - In vivo study of changes in refractive index distribution in the human crystalline lens with age and accommodation. AB - PURPOSE: Magnetic resonance imaging (MRI) was used to map the refractive index distribution in human eye lenses in vivo and to investigate changes with age and accommodation. METHODS: Whole-eye MR images were obtained for sagittal and transverse axial planes in one eye each of 15 young (19-29 years) and 15 older (60-70 years) subjects when viewing a far ( approximately 6 m) target and at individual near points in the young subjects. Refractive index maps of the crystalline lens were calculated by using a procedure previously validated in vitro. RESULTS: A central high refractive index plateau region and sharp decline in refractive index at the periphery were seen in all three groups. The peripheral decline was steepest in the older lenses and least steep in the young accommodated lenses. Average lens thickness increased (+0.27 mm; P < 0.05) and equatorial diameter decreased (-0.35 mm; P < 0.05) with accommodation. Axial thickness (+0.96 mm; P < 0.05) and equatorial diameter (+0.28 mm; P < 0.05) increased with age. The central index (1.409 +/- 0.008) did not differ between groups. The axial thickness of the central plateau increased with age (+0.83 mm; P < 0.05) but not significantly with accommodation. The equatorial diameter of the central plateau increased with age (+0.56 mm; P < 0.01) and decreased with accommodation (-0.43 mm; P < 0.05). CONCLUSIONS: The refractive index of the central plateau region does not change significantly with accommodation or ageing, but its size increases with age and the peripheral decline in refractive index becomes steeper in older lenses. PMID- 18408188 TI - Plus disease in retinopathy of prematurity: development of composite images by quantification of expert opinion. AB - PURPOSE: To demonstrate a methodology for generating composite wide-angle images of plus disease in retinopathy of prematurity (ROP), using quantitative analysis of expert opinions. METHODS: Thirty-four wide-angle retinal images were independently interpreted by 22 ROP experts as "plus" or "not plus." All images were processed by the computer-based Retinal Image multiScale Analysis (RISA) system to calculate two parameters: arterial integrated curvature (AIC) and venous diameter (VD). Using a reference standard defined by expert consensus, sensitivity and specificity curves were calculated by varying the diagnostic cutoffs for AIC and VD. From these curves, individual vessels from multiple images were identified with particular diagnostic cutoffs, and were combined into composite wide-angle images using graphics-editing software. RESULTS: The values associated with 75% underdiagnosis of true plus disease (i.e., 25% sensitivity cutoff) were AIC 0.061 and VD 4.272, the values associated with 50% underdiagnosis of true plus disease (i.e., a 50% sensitivity cutoff) were AIC 0.049 and VD 4.088, and the values associated with 25% underdiagnosis of true plus disease (i.e., 75% sensitivity cutoff) were AIC 0.042 and VD 3.795. Composite wide-angle images were generated by identifying and combining individual vessels with these characteristics. CONCLUSIONS: Computer-based image analysis permits quantification of retinal vascular features, and a spectrum of abnormalities is seen in ROP. Selection of appropriate vessels from multiple images can produce composite plus disease images corresponding to expert opinions. This method may be useful for educational purposes, and for development of future disease definitions based on objective, quantitative principles. PMID- 18408190 TI - Engrafted chicken neural tube-derived stem cells support the innate propensity for axonal regeneration within the rat optic nerve. AB - PURPOSE: Injury to the adult optic nerve, caused mechanically or by diseases, is still not reparable because the retinal ganglion cells (RGCs) are not allowed to regrow their axons and die retrogradely, although they possess the intrinsic propensity to regenerate axons in experimental conditions. METHODS: In vitro propagated embryonic stem cells derived from the early chicken neural tube (NTSCs) were used to examine whether transplanted NTSCs produce growth-promoting factors and pave the microenvironment, thus facilitating axonal regeneration within the rat optic nerve. RESULTS: NTSCs survived within the site where the optic nerve had been cut and continued to be nestin-positive, thus preserving their undifferentiated cell phenotype. Transplanted NTSCs activated the matrix metalloproteases (MMP)-2 and -14 in glial fibrillary acidic protein (GFAP) positive optic nerve astrocytes. MMP2 production correlated with immunohistochemically visible degradation of inhibitory chondroitin sulfate proteoglycans (CSPGs). In addition, NTSCs produced a panoply of neurite-promoting factors including oncomodulin, ciliary neurotrophic factor, brain-derived neurotrophic factor and crystallins beta and gamma. Cut axons intermingled with NTSCs and passed through the zone of injury to enter the distal optic nerve over long distances, arriving at the thalamus and midbrain. CONCLUSIONS: This study showed evidence that paving of the distal optic nerve microenvironment with proteolytically active MMPs and providing stem-cell-derived growth factors is a suitable method for facilitating regenerative repair of the optic nerve. Understanding the molecular mechanisms of this repair has fundamental implications for development of NTSC-based subsidiary therapy after neural injuries. PMID- 18408192 TI - Adult-onset myopia: the Genes in Myopia (GEM) twin study. AB - PURPOSE: To report the frequency of adult-onset myopia in a large cohort of Caucasian twins that were assessed as part of the Genes in Myopia (GEM) twin study and to quantify the genetic contribution in adult-onset myopia using the classic twin model. METHODS: All twins aged 18 years or older were invited to participate in the GEM twin study through the Australian Twin Registry (ATR). Each twin completed a standard questionnaire and underwent a comprehensive eye assessment, including cycloplegic objective examination. Adult-onset myopia was defined as having the first spectacle/contact lens correction at the age of 18 years or older. Myopia was defined as spherical equivalent worse than or equal to -0.50 D. RESULTS: A total of 1224 twins (690 monozygotic [MZ] and 534 dizygotic [DZ]) between 18 and 86 years of age were recruited into the GEM study. A total of 96 twins (96/347 = 27.7%) comprising 50 MZ and 46 DZ twins were first prescribed optical correction for myopia at the age of 18 years or older. A significantly higher MZ intrapair correlation (r = 0.61) compared with that in DZ twins (r = 0.16, P < 0.01) for spherical equivalent was found in twins with adult onset myopia. CONCLUSIONS: Adult-onset myopia is a relatively common condition, with approximately one quarter of cases occurring in adulthood. To the authors' knowledge, the GEM twin study is the first study of its kind to provide evidence to support a genetic component in adult-onset myopia. PMID- 18408191 TI - Light-induced retinal changes observed with high-resolution autofluorescence imaging of the retinal pigment epithelium. AB - PURPOSE: Autofluorescence fundus imaging using an adaptive optics scanning laser ophthalmoscope (AOSLO) allows for imaging of individual retinal pigment epithelial (RPE) cells in vivo. In this study, the potential of retinal damage was investigated by using radiant exposure levels that are 2 to 150 times those used for routine imaging. METHODS: Macaque retinas were imaged in vivo with a fluorescence AOSLO. The retina was exposed to 568- or 830-nm light for 15 minutes at various intensities over a square (1/2) degrees per side. Pre- and immediate postexposure images of the photoreceptors and RPE cells were taken over a 2 degrees field. Long-term AOSLO imaging was performed intermittently from 5 to 165 days after exposure. Exposures delivered over a uniform field were also investigated. RESULTS: Exposures to 568-nm light caused an immediate decrease in autofluorescence of RPE cells. Follow-up imaging revealed either full recovery of autofluorescence or long-term damage in the RPE cells at the exposure. The outcomes of AOSLO exposures and uniform field exposures of equal average power were not significantly different. No effects from 830-nm exposures were observed. CONCLUSIONS: The study revealed a novel change in RPE autofluorescence induced by 568-nm light exposure. Retinal damage occurred as a direct result of total average power, independent of the light-delivery METHOD: Because the exposures were near or below permissible levels in laser safety standards, these results suggest that caution should be used with exposure of the retina to visible light and that the safety standards should be re-evaluated for these exposure conditions. PMID- 18408193 TI - Antimicrobial efficacy of riboflavin/UVA combination (365 nm) in vitro for bacterial and fungal isolates: a potential new treatment for infectious keratitis. AB - PURPOSE: To demonstrate the antimicrobial properties of riboflavin/UVA (365 nm) against common pathogens. METHODS: One group of bacteria (Pseudomonas aeruginosa [PA], Staphylococcus aureus [SA], and Staphylococcus epidermidis [SE]) was tested by using Kirby-Bauer discs with (1) empty disc (Control - C); (2) riboflavin 0.1% (B2); (3) riboflavin 0.1% previously activated by UVA (B2'); (4) UVA alone (UVA); (5) group 2+additional UVA exposure (UVA+B2); and (6) group 3+additional UVA exposure (UVA+B2'). In addition, another group of microbes was tested with the same approach: methicillin-resistant S. aureus (MRSA), multidrug-resistant P. aeruginosa (MDRPA), drug-resistant Streptococcus pneumoniae (DRSP), and Candida albicans (CA). The mean growth inhibition zone (GIZ) in square millimeters was measured around the discs. The mean standard deviation (MSD) was calculated to be 3.65 when alpha = 0.01. A mean deviation (MD) > MSD indicates a significant difference. RESULTS: In the first group, the GIZ was significantly greater after UVA (MD = 14.30), UVA+B2 (MD = 39.61), and UVA+B2' (MD = 40.45) when compared with C, B2, and B2'. UVA alone was less effective than UVA+B2 (MD = 25.31) and UVA+B2' (MD = 26.15). The second group demonstrated increased GIZ in UVA (MD = 6.98), UVA+B2 (MD = 17.80), and UVA+B2' (MD = 21.15) when compared with C, B2, and B2'. UVA alone was less effective against the second group of bacteria than was UVA+B2 (MD = 10.82) and UVA+B2' (MD = 14.17). CA did not show any GIZ after treatment. CONCLUSIONS: Riboflavin/UVA was effective against SA, SE, PA, MRSA, MDRPA, and DRSP, but was ineffective on CA and has the potential for use in treatment of microbial keratitis in the future. PMID- 18408194 TI - Proline-glutamic acid-proline-lysine repetition peptide as an antigen for the serological diagnosis of strangles. AB - The reactivity of the proline-glutamic acid-proline-lysine (PEPK) repetition peptide antigen in 3176 serum samples was investigated to evaluate its utility as an antigen for the serological diagnosis of strangles. The reactivity of the sera of horses infected with Streptococcus equi subspecies equi was high when the peptide had several PEPK repetitions. However, as the number of PEPK repetitions increased, the reactivity of the antigen with the sera of horses infected with Streptococcus equi subspecies zooepidemicus also increased. In horses infected experimentally with S equi, the reactivity of the PEPK antigen with five repetitions increased one week after inoculation and continued to increase during the following four weeks. The optical density (OD) values of test sera from horses infected experimentally with S equi and sera from horses that had recovered from strangles were high. The od values of sera from horses that had recovered from an experimental infection with S zooepidemicus and of sera from healthy horses were comparatively low. PMID- 18408195 TI - Subclinical hypocalcaemia in captive Asian elephants (Elephas maximus). AB - The hypothesis that hypocalcaemia may play a role in dystocia in captive Asian elephants (Elephas maximus) was investigated. The objectives of the study were to measure the total calcium concentration in elephant plasma; assess the changes in parameters of calcium metabolism during a feeding trial; investigate a possible relationship between calcium metabolism and dystocia; and assess bone mineralisation in captive Asian elephants in vivo. The following parameters were measured: total and ionised calcium, inorganic phosphorous and magnesium, the fractional excretions of these minerals, intact parathyroid hormone, 25-OH-D(3) and 1,25-OH-D(3). Radiographs were taken from tail vertebrae for assessment of bone mineralisation. The mean (sd) heparinised plasma total calcium concentration was 2.7 (0.33) mmol/l (n=43) ranging from 0.84 to 3.08 mmol/l in 11 Asian elephants. There was no significant correlation between plasma total calcium concentration and age. Following feeding of a calcium rich ration to four captive Asian elephant cows, plasma total and ionised calcium peaked at 3.6 (0.24) mmol/l (range 3.4 to 3.9 mmol/l) and 1.25 (0.07) mmol/l (range 1.17 to 1.32 mmol/l), respectively. Plasma ionised calcium concentrations around parturition in four Asian elephant cows ranged from 0.37 to 1.1 mmol/l only. The present study indicates that captive Asian elephants might be hypocalcaemic, and that, in captive Asian elephants, the normal plasma concentration of total calcium should actually be around 3.6 mmol/l and normal plasma concentration of ionised calcium around 1.25 mmol/l. Given the fact that elephants absorb dietary calcium mainly from the intestine, it could be concluded that elephants should be fed calcium rich diets at all times, and particularly around parturition. In addition, normal values for ionised calcium in captive Asian elephants should be reassessed. PMID- 18408196 TI - Rupture of the dura mater in two dogs caused by the peracute extrusion of a cervical disc. AB - A rupture of the dura mater caused by the peracute extrusion of a cervical disc was diagnosed by myelography in two dogs. In both cases traction on the cervical spine resulted in contrast medium entering the ruptured intervertebral disc from the subarachnoid space. Both dogs became suddenly tetraparetic and unable to ambulate during vigorous exercise, but regained the ability to walk without surgical treatment. PMID- 18408197 TI - Eimeria species in cattle on farms in England and Wales. PMID- 18408198 TI - Antibody response of wild boar (Sus scrofa) piglets vaccinated against Aujeszky's disease virus. PMID- 18408199 TI - Cerebral and renal candidosis in a llama (Lama glama). PMID- 18408200 TI - Pulmonary haemorrhage and fibrillary glomerulonephritis (pulmonary-renal syndrome) in a dog. PMID- 18408201 TI - Cyanide poisoning associated with the feeding of apricot kernels to dairy cattle. PMID- 18408202 TI - Induced brain lesions in calves infected with bluetongue virus. PMID- 18408203 TI - Australian equine influenza: vaccine protection in the UK. PMID- 18408204 TI - Parascaris equorum resistance to moxidectin? PMID- 18408205 TI - Docked dogs at Crufts. PMID- 18408206 TI - Riding establishments inspectorate. PMID- 18408207 TI - Emotional intelligence of veterinary students. PMID- 18408208 TI - Our minds are made up--don't confuse us with the facts: commentary on policies concerning children with sexual behavior problems and juvenile sex offenders. AB - This commentary examines four common policy-relevant perceptions of teen and preteen sex offenders-high risk, "specialness," homogeneity, and intransigence. Each perception is contrasted with long-standing as well as more current scientific facts. It is argued that public policies for these youth have been fundamentally driven by misperceptions, resulting in a set of well-intentioned but ultimately flawed policies and practices that are unlikely to deliver either child protection or juvenile justice benefits. These include federal and state policies pertaining to public registration and notification, community management, institutional placement, treatment approaches, and treatment standards. The research evidence about these juveniles is considerably more positive than current policies or clinical practices might suggest, and reflects a sharp disconnect between popular policy-relevant perceptions and the facts as we know them about these diverse cases. PMID- 18408209 TI - Beyond sexual abuse: the impact of other maltreatment experiences on sexualized behaviors. AB - This study sought to broaden research findings linking maltreatment to sexualized behaviors by investigating whether maltreatment experiences other than sexual abuse predict such behaviors. The sample included 690 children without reported sexual abuse histories who are participants in the LONGSCAN Consortium, a prospective multisite investigation of childhood maltreatment. Child Protective Service reports before age 8 years and caregiver reports on the Child Sexual Behavior Inventory-II at age 8 years were used to examine the relationship between maltreatment timing and type, and sexualized behaviors. Logistic regression analyses suggested that early (< 4) and late (4-8) reports of physical abuse were associated with more sexualized behaviors (odds ratios = 1.9-2.6). The pattern differed by gender, with physical abuse predicting sexual intrusiveness and displaying private parts in boys, and boundary problems in girls. Findings suggest that maltreatment other than sexual abuse, and the developmental periods in which it occurs, may be linked to the development of sexualized behaviors. PMID- 18408210 TI - Treatment outcome and criminal offending by youth with sexual behavior problems. AB - Children and adolescents treated for general delinquency problems and rated by caregivers as having sexual behavior problems (SBP; N = 696) were compared with youth from the same sample with no sexual behavior problems (NSBP; N = 1,185). Treatment outcome through 12-months posttreatment and criminal offending through an average 48-month posttreatment were compared for both groups. It was hypothesized that both groups would improve over time; however, the SBP group would evidence greater psychopathology at follow-up, and these hypotheses were supported. It was further hypothesized that youth with SBP would not differ from youth with NSBP in rates of future sexual or nonsexual offenses. These hypotheses were also supported. SBP group membership was not a significant predictive factor in analyses modeling future offending (any) or future person offenses. Few youth in either group had sexual offenses. The importance of these findings for clinical and policy decision making is discussed. PMID- 18408211 TI - Meta-analysis of treatment for child sexual behavior problems: practice elements and outcomes. AB - This meta-analysis of 11 treatment outcome studies evaluated 18 specific treatments of sexual behavior problems (SBP) as a primary or secondary target. Specifically, it examines relations among child characteristics, treatment characteristics (including practice elements), and short-term outcome (including sexual and general behavior problems). Utilizing pre- and postintervention results, the overall degree of change over the course of treatment was estimated at a 0.46 and 0.49 standard deviation decline in SBP and general behavior problems, respectively. As hypothesized, the caregiver practice element Parenting/Behavior Management Skills (BPT) predicted the Child Sexual Behavior Inventory (and the Child Behavior Checklist when BPT was combined with caregiver Rules about Sexual Behaviors). In contrast, practice elements that evolved from Adult Sex Offender (ASO) treatments were not significant predictors. BPT and preschool age group provided the best model fit and more strongly predicted outcome than broad treatment type classifications (e.g., Play Therapy or Cognitive Behavior Therapy). Results question current treatments for children with SBP that are based on ASO models of treatment without caregiver involvement. PMID- 18408212 TI - Sexual anxiety and eroticism predict the development of sexual problems in youth with a history of sexual abuse. AB - Youth with confirmed histories of sexual abuse (N = 118) were followed longitudinally to examine associations between their initial sexual reactions to abuse and subsequent sexual functioning. Participants were interviewed at abuse discovery (ages 8 through 15) and again 1 and 6 years later. Eroticism and sexual anxiety emerged as distinct indices of abuse-specific sexual reactions and predicted subsequent sexual functioning. Eroticism was associated with indicators of heightened sexuality, including more sexual risk behavior and views of sexual intimacy focused on partners' needs. Sexual anxiety was associated with indicators of diminished sexuality, including few sexual partners and avoidant views of sexual intimacy. Age at abuse discovery moderated some associations, suggesting that the timing of abuse-specific reactions affects trajectories of sexual development. Findings point to the need for a developmental approach to understanding how abuse-specific sexual reactions disrupt sexual development and the need for early interventions promoting healthy sexual development. PMID- 18408213 TI - Predictors of problematic sexual behavior among children with complex maltreatment histories. AB - Exploratory analyses of sexual behavior problems (SBP) were conducted within a larger epidemiological study of 347 preadolescent children in foster and kinship care. SBP was estimated from carergiver-reported scores on the Assessment Checklist for Children. The study simultaneously examined a large number of discrete and cumulative influences on the development of children at high risk for SBP. Most children with SBP had corresponding psychopathology, most notably conduct problems, inattention, and interpersonal behavior problems suggestive of attachment disturbances. Several correlates identified in previous studies were not associated with SBP. High concordance of SBP was found among 52 sibling dyads. Independent predictors of SBP were older age at entry into care, female gender, placement instability, and contact sexual abuse. The findings emphasize the significance of cumulative risk among children exposed to multiple adversities. The findings generated several hypothesized mechanisms involving attachment disturbances. PMID- 18408214 TI - Report of the ATSA Task Force on Children With Sexual Behavior Problems. PMID- 18408215 TI - In memoriam: William N. Friedrich, 1951-2005. PMID- 18408216 TI - Mitotic phosphorylation stimulates DNA relaxation activity of human topoisomerase I. AB - Human DNA topoisomerase I (topo I) is an essential mammalian enzyme that regulates DNA supercoiling during transcription and replication. In addition, topo I is specifically targeted by the anticancer compound camptothecin and its derivatives. Previous studies have indicated that topo I is a phosphoprotein and that phosphorylation stimulates its DNA relaxation activity. The locations of most topo I phosphorylation sites have not been identified, preventing a more detailed examination of this modification. To address this issue, mass spectrometry was used to identify four topo I residues that are phosphorylated in intact cells: Ser(10), Ser(21), Ser(112), and Ser(394). Immunoblotting using anti phosphoepitope antibodies demonstrated that these sites are phosphorylated during mitosis. In vitro kinase assays demonstrated that Ser(10) can be phosphorylated by casein kinase II, Ser(21) can be phosphorylated by protein kinase Calpha, and Ser(112) and Ser(394) can be phosphorylated by Cdk1. When wild type topo I was pulled down from mitotic cells and dephosphorylated with alkaline phosphatase, topo I activity decreased 2-fold. Likewise, topo I polypeptide with all four phosphorylation sites mutated to alanine exhibited 2-fold lower DNA relaxation activity than wild type topo I after isolation from mitotic cells. Further mutational analysis demonstrated that Ser(21) phosphorylation was responsible for this change. Consistent with these results, wild type topo I (but not S21A topo I) exhibited increased sensitivity to camptothecin-induced trapping on DNA during mitosis. Collectively these results indicate that topo I is phosphorylated during mitosis at multiple sites, one of which enhances DNA relaxation activity in vitro and interaction with DNA in cells. PMID- 18408217 TI - Lysophosphatidic acid protects cancer cells from histone deacetylase (HDAC) inhibitor-induced apoptosis through activation of HDAC. AB - Histone deacetylases (HDACs) catalyze the removal of acetyl groups from histones and contribute to transcriptional repression. In addition, the HDAC inhibitors induce apoptosis in cancer cells through alterations in histone acetylation and activation of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) apoptotic pathway. Lysophosphatidic acid (LPA) is a growth factor that promotes survival of cancer cells through activation of G protein-coupled receptors. Here we show that HDAC inhibitors can induce apoptosis through activation of the TRAIL apoptotic pathway, and LPA prevented HDAC inhibitor-induced apoptosis and increased TRAIL receptor DR4 (death receptor 4) protein expression. This was associated with increased HDAC1 recruitment to the DR4 promoter following LPA treatment and a reduction in HDAC inhibitor-induced histone acetylation in the DR4 promoter. In addition, LPA induces HDAC enzyme activity in a dose- and time dependent manner, and this is associated with HDAC1 activation and increased binding of HDAC1 to HDAC2. Reducing the expression of HDAC1 significantly lowered LPA-induced HDAC activity and increased histone acetylation. LPA induction of HDAC activity was blocked by the LPA receptor antagonist, Ki16425, or by inhibiting receptor activation with pertussis toxin. Reducing the expression of the LPA receptor LPA(1) also blocked LPA-induced HDAC activation. In addition, LPA reduced histone acetyltransferase enzymatic activity. Finally, LPA attenuated the ability of the HDAC inhibitor to reduce HDAC activity. Thus, LPA enhances survival of cancer cells by increasing HDAC activity and reducing histone acetylation. PMID- 18408218 TI - Transtheoretical model-based dietary interventions in primary care: a review of the evidence in diabetes. AB - The objective of this study was to review the evidence concerning stage-based dietary interventions in primary care among persons with diabetes or an elevated diabetes risk. Search strategies were electronic databases and manual search. Selection criteria were randomized controlled studies with stage-based dietary intervention, conducted in primary care with at least 6 months of follow-up, and participants with either type 2 diabetes or with an elevated risk. The researchers evaluated trials for inclusion, extracted data and assessed study quality. Seven articles, based on five data sets, were included. These studies concentrated on cardiovascular diseases and being overweight, not diabetes. The quality of the studies was moderate to weak. Inadequacies in the reporting often involved unspecific information on the training of health care providers. Long term positive outcomes (compared with controls) were found in total and monounsaturated fat intake, diastolic blood pressure, health status and well being. The existing data are insufficient for drawing conclusions on the benefits of the transtheoretical model. More high-quality studies focusing on diabetes are needed, with greater attention to the training of providers and process evaluation. There is a need for a standardized appraisal tool for study evaluation, focusing separately on education interventions for patients and providers. PMID- 18408219 TI - Leaf succulence determines the interplay between carboxylase systems and light use during Crassulacean acid metabolism in Kalanchoe species. AB - The photosynthetic physiology of Crassulacean acid metabolism was investigated in two Kalanchoe species with differing leaf succulence. The magnitude of CAM was higher for the more succulent leaves of K. daigremontiana, compared to the less succulent leaves of K. pinnata. High succulence was related to low mesophyll conductance: K. pinnata was able to maximize diurnal carbon gain by the C(3) pathway, whereas increased succulence is associated with a higher commitment to the CAM cycle in K. daigremontiana. The Rubisco specificity factor, tau, determining selectivity for CO(2) over O(2), was similar for both species (approximately 88), and lower than that of Spinacea (approximately 95), but in contrast to C(4) plants, the Rubisco K(mCO(2)) (determined independently) was also lower in Kalanchoe spp. than in spinach. Differences in light use were related to the nature of the sink strength in each Phase of CAM, with PEPC activity resulting in low electron transport rates. Decarboxylation was marked by high, non-saturated rates of electron transport, with Rubisco activity and activation state increasing in both species during the course of the light period. The degree of succulence, and extent of CAM activity, was associated with a progressive inhibition of PSII photochemistry and potential Rubisco activity during the night in both species. Rubisco could be 'woken up' more rapidly in K. pinnata, whereas 45 min light acclimation was required for full recovery of electron transport and Rubisco activity in K. daigremontiana. Leaf morphology therefore seems to alter the expression of and dependence on CAM, but also the extent of co-regulation of carboxylase networks and light use capacity. PMID- 18408220 TI - Cloning, localization and expression analysis of vacuolar sugar transporters in the CAM plant Ananas comosus (pineapple). AB - In photosynthetic tissues of the CAM plant pineapple (Ananas comosus), storage of soluble sugars in the central vacuole during the daytime and their remobilization at night is required to provide carbon skeletons for nocturnal CO(2) fixation. However, soluble sugars produced photosynthetically must also be exported to support growth processes in heterotrophic tissues. To begin to address how vacuolar sugar storage and assimilate partitioning are regulated in A. comosus, degenerate PCR and cDNA library screening were used to clone three candidate sugar transporters from the leaves of this species. Subcellular localization of the three transporters was investigated via expression of YFP-fusion proteins in tobacco epidermal cells and their co-localization with subcellular markers by confocal microscopy. Using this strategy, a putative hexose transporter (AcMST1) and a putative inositol transporter (AcINT1) were identified that both localized to the tonoplast, whereas a putative sucrose transporter (AcSUT1) was found to localize to prevacuolar compartments. A cDNA (AcMST2) with high similarity to a recently characterized tonoplast hexose transporter in Arabidopsis was also identified from an A. comosus fruit EST database. Analyses of transcript abundance indicated that AcMST1 was more highly expressed in fruits compared to leaves of A. comosus, whilst transcripts of AcINT1, AcSUT1, and AcMST2 were more abundant in leaves. Transcript abundance of AcINT1, the putative inositol transporter, showed day-night changes comparable to those of other CAM-related transcripts described in Mesembryanthemum crystallinum. The results are discussed in terms of the role of vacuolar sugar transporters in regulating carbon flow during the diel cycle in CAM plants. PMID- 18408221 TI - Maize C4-form phosphoenolpyruvate carboxylase engineered to be functional in C3 plants: mutations for diminished sensitivity to feedback inhibitors and for increased substrate affinity. AB - Introducing a C(4)-like pathway into C(3) plants is one of the proposed strategies for the enhancement of photosynthetic productivity. For this purpose it is necessary to provide each component enzyme that exerts strong activity in the targeted C(3) plants. Here, a maize C(4)-form phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) was engineered for its regulatory and catalytic properties so as to be functional in the cells of C(3) plants. Firstly, amino acid residues Lys 835 and Arg-894 of maize PEPC, which correspond to Lys-773 and Arg-832 of Escherichia coli PEPC, respectively, were replaced by Gly, since they had been shown to be involved in the binding of allosteric inhibitors, malate or aspartate, by our X-ray crystallographic analysis of E. coli PEPC. The resulting mutant enzymes were active but their sensitivities to the inhibitors were greatly diminished. Secondly, a Ser residue (S780) characteristically conserved in all C(4)-form PEPC was replaced by Ala conserved in C(3)- and root-form PEPCs to decrease the half-maximal concentration (S(0.5)) of PEP. The double mutant enzyme (S780A/K835G) showed diminished sensitivity to malate and decreased S(0.5)(PEP) with equal maximal catalytic activity (V(m)) to the wild-type PEPC, which will be quite useful as a component of the C(4)-like pathway to be introduced into C(3) plants. PMID- 18408222 TI - Neutrophil role in in vivo anti-lymphoma activity of rituximab: FCGR3B-NA1/NA2 polymorphism does not influence response and survival after rituximab treatment. AB - BACKGROUND: Neutrophils could play an important role in in vivo rituximab anti lymphoma activity. FcgammaRIIIb is expressed only by neutrophils and FcgammaRIIIb neutrophil antigen (NA)1/NA2 polymorphism influenced phagocytosis of immunoglobulin G1-opsonized particles. We formulated the hypothesis that if neutrophils are critical cells for in vivo rituximab activity, FcgammaRIIIb NA1/NA2 polymorphism could influence the response to rituximab. PATIENTS AND METHODS: FCGR3B-NA1/NA2 genotypes were determined in 46 patients having received rituximab for a previously untreated, follicular, non-Hodgkin's lymphoma. The clinical response and the disappearance of the BCL2-JH gene rearrangement in both peripheral blood and bone marrow were evaluated at 2 months (M2) and each year during 7 years. RESULTS: They were 13% homozygous for FCGR3B-NA1, 61% homozygous for FCGR3B-NA1/NA2 and 26% heterozygous. The objective response rates at M2 were 67% in homozygous FCGR3B-NA1 patients compared with 75% in homozygous FCGR3B-NA2 and 75% in heterozygous patients (not significant). We found no difference for progression-free and overall survival by FCGR3B-NA1/NA2 genotypes. CONCLUSION: These results indicate no association between FCGR3B-NA1/NA2 polymorphism and response to rituximab indicating no significant role of phagocytosis mediated by neutrophils in in vivo mechanism of rituximab activity. PMID- 18408223 TI - Survival and reproductive function of 52 women treated with surgery and bleomycin, etoposide, cisplatin (BEP) chemotherapy for ovarian yolk sac tumor. AB - BACKGROUND: Ovarian yolk sac tumor (YST) is a very rare malignancy arising in young women. Chemotherapy has dramatically improved the prognosis. Current treatment consists of surgery followed by bleomycin, etoposide, and cisplatin (BEP) chemotherapy. However, given the rarity of this tumor, ovarian YST-specific survival and outcome after such treatment are not precisely known. PATIENTS AND METHODS: This report concerns prospectively recorded cases that were either treated at Institut Gustave Roussy (Villejuif, France) or referred there for advice about therapy. From 1990 to 2006, 52 patients underwent surgery followed by BEP chemotherapy. Data on patient characteristics, treatment, survival, and fertility outcome were analyzed to assess treatment efficacy and gonadal toxicity after achieving a complete remission. RESULTS: Thirty-five patients had stage I/II tumors while 17 patients presented with stage III/IV disease. With a median follow-up of 68 months, the overall 5-year survival and disease-free survival rates were 94% and 90%, respectively. Forty-one women underwent fertility-sparing surgery. Pregnancy was achieved in 12 of 16 (75%) women who attempted conception. Overall, 19 pregnancies have been recorded. CONCLUSIONS: BEP chemotherapy following fertility-sparing surgery is a very effective treatment of ovarian YSTs. Most of the patients who attempt conception after complete remission will have children. PMID- 18408224 TI - First-line bevacizumab combined with reduced dose interferon-alpha2a is active in patients with metastatic renal cell carcinoma. AB - BACKGROUND: In patients with untreated metastatic renal cell carcinoma (mRCC), progression-free survival (PFS) was longer with bevacizumab + interferon (IFN) alpha than IFN + placebo (AVOREN trial). In this hypothesis-generating study, subgroup analysis was carried out to determine the effect of IFN dose reduction. PATIENTS AND METHODS: A total of 649 patients received IFN 9 MIU s.c. three times weekly plus bevacizumab 10 mg/kg or placebo every 2 weeks until disease progression. The IFN dose was reduced to 6 or 3 MIU with the development of IFN attributed toxicity. Differences between treatment arms in PFS, response rate and tolerability were analysed in the reduced-dose group. RESULTS: IFN dose was reduced in 131 patients in the bevacizumab + IFN arm and 97 patients in the IFN + placebo arm during the trial. PFS rates in the bevacizumab + reduced-dose IFN group were comparable with the total population (Kaplan-Meier estimates of event free rate at 1 year: 0.524 versus 0.427). Bevacizumab + reduced-dose IFN was well tolerated, with substantial decreases in the rate of adverse events following dose reduction. CONCLUSION: This retrospective subgroup analysis suggests that the dose of IFN can be reduced to manage side-effects while maintaining efficacy in patients with mRCC receiving bevacizumab + IFN. PMID- 18408226 TI - Insulin-like growth factor 1, insulin-like growth factor-binding protein 3, and testicular germ-cell tumor risk. AB - Studies have consistently shown that taller men are at increased risk of testicular germ-cell tumors. Thus, it is plausible that factors associated with height may also influence risk of these tumors. The authors examined associations between testicular germ-cell tumor risk and circulating concentrations of insulin like growth factor 1 (IGF-1) and insulin-like growth factor-binding protein 3 (IGFBP-3) among 517 cases and 790 controls from the US Servicemen's Testicular Tumor Environmental and Endocrine Determinants (STEED) Study (2002-2005). Odds ratios and 95% confidence intervals were estimated using logistic regression models, adjusting for age, race/ethnicity, height, and body mass index. All tests of significance were two-sided. Overall, there were no associations between IGF-1 or IGFBP-3 concentrations and risk of testicular germ-cell tumors (p > 0.05). However, when cases were separated by histologic type, there was a suggestion of a reduction in seminoma risk associated with the highest concentrations of IGF-1 as compared with the lowest concentrations (odds ratio = 0.66, 95% confidence interval: 0.40, 1.09). Although there were no overall associations with insulin like growth factor, contrary to expectation, there was a suggestion that IGF-1 concentrations may be inversely associated with risk of seminoma. PMID- 18408225 TI - Personal use of hair dye and the risk of certain subtypes of non-Hodgkin lymphoma. AB - Personal use of hair dye has been inconsistently linked to risk of non-Hodgkin lymphoma (NHL), perhaps because of small samples or a lack of detailed information on personal hair-dye use in previous studies. This study included 4,461 NHL cases and 5,799 controls from the International Lymphoma Epidemiology Consortium 1988-2003. Increased risk of NHL (odds ratio (OR) = 1.3, 95% confidence interval (CI): 1.1, 1.4) associated with hair-dye use was observed among women who began using hair dye before 1980. Analyses by NHL subtype showed increased risk for follicular lymphoma (FL) and chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) but not for other NHL subtypes. The increased risks of FL (OR = 1.4, 95% CI: 1.1, 1.9) and CLL/SLL (OR = 1.5, 95% CI: 1.1, 2.0) were mainly observed among women who started using hair dyes before 1980. For women who began using hair dye in 1980 or afterward, increased FL risk was limited to users of dark-colored dyes (OR = 1.5, 95% CI: 1.1, 2.0). These results indicate that personal hair-dye use may play a role in risks of FL and CLL/SLL in women who started use before 1980 and that increased risk of FL among women who started use during or after 1980 cannot be excluded. PMID- 18408227 TI - Parental subfecundity and risk of decreased semen quality in the male offspring: a follow-up study. AB - A few studies have found poor semen quality in sons whose mothers have received fertility treatment, but it is unknown whether the poor semen quality is related to the infertility treatment or to infertility per se, for example, whether it is caused by hereditable factors. Using data from a population-based, Danish follow up study conducted in 2005-2006, the authors of the present study examined whether sons of subfertile couples who had not received fertility treatment had poorer semen quality than sons of fertile couples. Among the 311 participants, an inverse association between parental waiting time to pregnancy and both semen volume and total sperm count was observed (p trend = 0.04 and p trend = 0.046, respectively). Semen volume in sons of subfertile parents (pregnant after > or =1 years) was 19% lower in comparison with that in sons of parents whose waiting time to pregnancy was 0-6 months (p = 0.02). Additionally, sperm concentration and percentage of morphologically normal sperm were, respectively, 22% (p = 0.15) and 23% (p = 0.13) lower in sons of subfertile parents. Results suggest a small to-moderate effect of parental subfecundity on semen quality in sons, comparable with the hypothesis that low fecundity has at least partly hereditable causes. PMID- 18408228 TI - Does temperature modify the association between air pollution and mortality? A multicity case-crossover analysis in Italy. AB - Adverse health effects of particulate matter <10 microm in aerodynamic diameter (PM(10)) and high temperatures are well known, but the extent of their interaction on mortality is less clear. This paper describes effect modification of temperature in the PM(10)-mortality association and tests the hypothesis that higher PM(10) effects in summer are due to enhanced exposure to particles. All deaths of residents of nine Italian cities between 1997 and 2004 were selected. The case-crossover approach was adopted to estimate the effect of PM(10) on mortality by season and temperature level. Three strata of temperature corresponding to low, medium, and high "ventilation" were identified, and the interaction between PM(10) and temperature within each stratum was examined. Season and temperature levels strongly modified the PM(10)-mortality association: for a 10-microg/m(3) variation in PM(10), a 2.54% increase in risk of death in summer (95% confidence interval: 1.31, 3.78) compared with 0.20% (95% confidence interval: -0.08, 0.49) in winter. Analysis of the interaction between PM(10) and temperature within temperature strata resulted in positive but, in most cases, nonstatistically significant coefficients. The authors found much higher PM(10) effects on mortality during warmer days. The hypothesis that such an effect is attributable to enhanced exposure to particles in summer could not be rejected. PMID- 18408229 TI - A review of the fetal brain cytokine imbalance hypothesis of schizophrenia. AB - Maternal infection during pregnancy increases the risk of schizophrenia and other brain disorders of neurodevelopmental origin in the offspring. A multitude of infectious agents seem to be involved in this association. Therefore, it has been proposed that factors common to the immune response to a wide variety of bacterial and viral pathogens may be the critical link between prenatal infection and postnatal brain and behavioral pathology. More specifically, it has been suggested that the maternal induction of pro-inflammatory cytokines may mediate the neurodevelopmental effects of maternal infections. Here, we review recent findings from in vitro and in vivo investigations supporting this hypothesis and further emphasize the influence of enhanced anti-inflammatory cytokine signaling on early brain development. Disruption of the fetal brain balance between pro- and anti-inflammatory cytokine signaling may thus represent a key mechanism involved in the precipitation of schizophrenia-related pathology following prenatal maternal infection and innate immune imbalances. PMID- 18408231 TI - Using health promotion outcomes in formative evaluation studies to predict success factors in interventions: an application to an intervention for promoting physical activity in Dutch children (JUMP-in). AB - JUMP-in is a systematically developed intervention aimed at promoting physical activity among primary school children. It is a joint project involving different authorities and entails six school-based programme components. Measuring effects of such an intervention is a complex challenge. A common problem is the lack of valid instruments to measure physical activity and its determinants. In addition, it usually takes years to find improvements in physical activity and related constructs like weight and fitness, or even in causal factors. For this reason different authors advocate for the establishment of 'health promotion outcomes'; (i) health literacy, (ii) social action and influence and (iii) healthy public policy and organizational practice. It is presumed that these health promotion outcomes lead to changes in determinants, behaviour and finally in health. Insight in these health promotion outcomes and information about input and through-put are important in discussing the impact and output. The formative evaluation study of the JUMP-in pilot shows the health promotion outcomes of the intervention. The health promotion outcomes 'social action and influence' and 'healthy public policy and organizational practices' were found to be positive. By measuring the presence of the conditions to achieve 'health literacy', it became clear that more attention must be paid to implementation in the future. Based on the health promotion outcomes, we expect that JUMP-in will be an effective intervention in the future. PMID- 18408232 TI - Smooth skyride through a rough skyline: Bayesian coalescent-based inference of population dynamics. AB - Kingman's coalescent process opens the door for estimation of population genetics model parameters from molecular sequences. One paramount parameter of interest is the effective population size. Temporal variation of this quantity characterizes the demographic history of a population. Because researchers are rarely able to choose a priori a deterministic model describing effective population size dynamics for data at hand, nonparametric curve-fitting methods based on multiple change-point (MCP) models have been developed. We propose an alternative to change-point modeling that exploits Gaussian Markov random fields to achieve temporal smoothing of the effective population size in a Bayesian framework. The main advantage of our approach is that, in contrast to MCP models, the explicit temporal smoothing does not require strong prior decisions. To approximate the posterior distribution of the population dynamics, we use efficient, fast mixing Markov chain Monte Carlo algorithms designed for highly structured Gaussian models. In a simulation study, we demonstrate that the proposed temporal smoothing method, named Bayesian skyride, successfully recovers "true" population size trajectories in all simulation scenarios and competes well with the MCP approaches without evoking strong prior assumptions. We apply our Bayesian skyride method to 2 real data sets. We analyze sequences of hepatitis C virus contemporaneously sampled in Egypt, reproducing all key known aspects of the viral population dynamics. Next, we estimate the demographic histories of human influenza A hemagglutinin sequences, serially sampled throughout 3 flu seasons. PMID- 18408230 TI - Structural cerebral variations as useful endophenotypes in schizophrenia: do they help construct "extended endophenotypes"? AB - Endophenotypes represent intermediate phenotypes on the putative causal pathway from the genotype to the phenotype. They offer a potentially valuable strategy to examine the molecular etiopathology of complex behavioral phenotypes such as schizophrenia. Neurocognitive and neurophysiological impairments that suggest functional impairments associated with schizophrenia have been proposed as endophenotypes. However, few studies have examined the structural variations in the brain that might underlie the functional impairments as useful endophenotypes for schizophrenia. Over the past three decades, there has been an impressive body of literature supporting brain structural alterations in schizophrenia. We critically reviewed the extant literature on the neuroanatomical variations in schizophrenia in this paper to evaluate their candidacy as endophenotypes and how useful they are in furthering the understanding of etiology and pathophysiology of schizophrenia. Brain morphometric measures meet many of the criteria set by different investigators, such as being robustly associated with schizophrenia, heritable, quantifiable, and present in unaffected family members more frequently than in the general population. We conclude that the brain morphometric alterations appear largely to meet the criteria for endophenotypes in psychotic disorders. Some caveats for the utility of endophenotypes are discussed. A proposal to combine more than one endophenotype ("extended endophenotype") is suggested. Further work is needed to examine how specific genes and their interactions with the environment may produce alterations in brain structure and function that accompany psychotic disorders. PMID- 18408233 TI - Recurrent tandem gene duplication gave rise to functionally divergent genes in Drosophila. AB - Tandem gene duplication is one of the major gene duplication mechanisms in eukaryotes, as illustrated by the prevalence of gene family clusters. Tandem duplicated paralogs usually share the same regulatory element, and as a consequence, they are likely to perform similar biological functions. Here, we provide an example of a newly evolved tandem duplicate acquiring novel functions, which were driven by positive selection. CG32708, CG32706, and CG6999 are 3 clustered genes residing in the X chromosome of Drosophila melanogaster. CG6999 and CG32708 have been examined for their molecular population genetic properties (Thornton and Long 2005). We further investigated the evolutionary forces acting on these genes with greater sample sizes and a broader approach that incorporate between-species divergence, using more variety of statistical methods. We explored the possible functional implications by characterizing the tissue specific and developmental expression patterns of these genes. Sequence comparison of species within D. melanogaster subgroup reveals that this 3-gene cluster was created by 2 rounds of tandem gene duplication in the last 5 Myr. Based on phylogenetic analysis, CG32708 is clearly the parental copy that is shared by all species. CG32706 appears to have originated in the ancestor of Drosophila simulans and D. melanogaster about 5 Mya, and CG6999 is the newest duplicate that is unique to D. melanogaster. All 3 genes have different expression profiles, and CG6999 has in addition acquired a novel transcript. Biased polymorphism frequency spectrum, linkage disequilibrium, nucleotide substitution, and McDonald-Kreitman analyses suggested that the evolution of CG6999 and CG32706 were driven by positive Darwinian selection. PMID- 18408235 TI - Antiretroviral therapy of late presenters with advanced HIV disease. AB - Potent antiretroviral therapy (ART) has dramatically improved the prognosis of HIV-1-infected individuals. However, 10% to 30% of patients in Western countries still present late for care, when CD4 T cells are below 200 cells/mm(3) and symptomatic HIV disease has occurred. Clinical considerations for advanced HIV disease are paramount as morbidity and mortality are directly correlated with a low initial CD4 T cell count, which is commonly associated with the simultaneous occurrence of co-morbidities, particularly opportunistic infections. Upon start of ART, the clinical entity of immune reconstitution inflammatory syndrome may occur and, in this context, raise the question of early versus delayed ART in patients treated for opportunistic infections. Recent data clearly indicate that an earlier start of ART is warranted in this latter situation. Guidelines for specific antiretroviral treatment for late-presenting patients are lacking. Knowledge about drug-drug interactions and co-morbidities should guide treatment choices and influence the clinical management and monitoring of drug-related side effects and interactions. Importantly, the outlook of patients who present late is very much dependent upon the initial response to ART. Nevertheless, even if optimal response to treatment has been achieved, long-term prognosis may be impaired in patients who initially presented with advanced HIV disease. We encourage physicians to perform HIV testing more frequently in order to detect HIV-infected individuals in time. PMID- 18408234 TI - Interactions of antimicrobial compounds with cross-linking agents of alginate dressings. AB - OBJECTIVES: The aim of this study was to investigate the antimicrobial activities of calcium ions and other cross-linking agents of alginate dressings, as well as their compatibility with commonly used topical antimicrobials. METHODS: The antimicrobial activities of cross-linking agents and antimicrobials (five antibiotics and four antiseptics) were evaluated by the broth dilution method. The interactions between individual cross-linking agents and antimicrobials were evaluated using the chequerboard test against common skin pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. RESULTS: From the MIC determined, antibiotics were the most active, followed by the antiseptics and cross-linking agents. Calcium ions, which are commonly used to cross-link alginate, exhibited very weak antimicrobial activity and higher fractional inhibitory concentration than the other cross-linking agents. The use of calcium and gentamicin resulted in antagonism against S. aureus. In contrast, aluminium, zinc and copper ions exhibited higher antimicrobial activities but insignificant interactions with the antimicrobials. CONCLUSIONS: Commonly used topical antimicrobials that are active against the skin pathogens S. aureus and P. aeruginosa could be potentially incompatible with calcium alginate dressings. Copper, zinc and aluminium ions are more suitable cross-linking agents for alginate as they do not show antagonism with the antimicrobials and could impart antimicrobial property to the resultant dressing. PMID- 18408236 TI - Successful oral therapy switch to trimethoprim/sulfamethoxazole in the case of an Enterococcus faecium liver abscess. PMID- 18408237 TI - Characterization of the inhibitory effect of voriconazole on the fungicidal activity of amphotericin B against Candida albicans in an in vitro kinetic model. AB - OBJECTIVES: The aim of the present investigation was to study and characterize the effect of voriconazole on the fungicidal activity of amphotericin B. METHODS: Four strains of Candida albicans susceptible to voriconazole were exposed to voriconazole and amphotericin B, either alone, simultaneously or sequentially in an in vitro kinetic model. Bolus doses resulting in voriconazole and amphotericin B concentrations of 0.005-5 and 2.5 mg/L, respectively, were administered. Antifungal-containing RPMI 1640 was eliminated and replaced by a fresh medium using a peristaltic pump, with a flow rate adjusted to obtain the desired half lives. With two drugs tested, a computer-controlled dosing pump compensated for differences in the elimination rates. Using static time-kill methodology, one C. albicans strain was exposed to 5 mg/L voriconazole for varying durations followed by 2.5 mg/L amphotericin B after three repeated washes of voriconazole. RESULTS: Voriconazole and amphotericin B treatment alone resulted in fungistatic and fungicidal activities, respectively. Simultaneous administration of voriconazole and amphotericin B resulted in fungicidal activity, whereas only fungistatic activity was observed when repeated doses of amphotericin B were administered sequentially after voriconazole at 24-96 h. The inhibition of the fungicidal activity of amphotericin B was voriconazole dose-dependent, but seemed to be recovered once the voriconazole concentration fell below the MIC. The fungicidal activity was quickly regained after the removal of voriconazole, irrespective of the duration of voriconazole pre-exposure. CONCLUSIONS: Voriconazole inhibited the fungicidal effect of sequentially administered amphotericin B in a concentration- and time-dependent manner; the clinical significance of this needs further investigation. PMID- 18408238 TI - Trimethoprim and enterococci in urinary tract infections: new perspectives on an old issue. AB - The lack of oral treatment alternatives for enterococcal urinary tract infections (UTIs) has led to a renewed interest in trimethoprim. Enterococci can incorporate exogenously produced folates and thereby reverse the effect of trimethoprim. Although a large proportion of enterococci appear susceptible to trimethoprim in vitro using standard media devoid of folates, a 360-fold increase in the MIC can be seen when susceptibility testing is performed in media containing fresh urine. Even if trimethoprim has a favourable pharmacokinetic profile, with high serum and very high urine concentrations, pharmacodynamic (PD) estimates show that a large proportion of the apparent wild-type isolates (as categorized by standard susceptibility testing) have unfavourable PD indices. The clinical efficacy of trimethoprim in enterococcal UTI is debated. We could identify not more than 38 evaluable cases of enterococcal UTI in the literature. The eradication rate was 82%. Case reports where patients on co-trimoxazole for UTI have developed bacteraemia with enterococci susceptible to trimethoprim seem to support experimental findings that standard antimicrobial susceptibility testing poorly predicts the clinical outcome of trimethoprim therapy. The European Committee on Antimicrobial Susceptibility Testing and the national breakpoint committees in Europe have recently debated the role of trimethoprim in the treatment of enterococcal UTI and agreed to categorize wild-type enterococci as intermediate to trimethoprim and trimethoprim/sulfamethoxazole. This allows the distinction between enterococci with and without acquired resistance mechanisms to trimethoprim. This review discusses the microbiological, experimental, clinical and PD aspects of the usage of trimethoprim for enterococcal UTI. PMID- 18408239 TI - An outpatient parenteral antibiotic therapy (OPAT) map to identify risks associated with an OPAT service. AB - OBJECTIVES: Administering parenteral antibiotics outside the confines of a ward setting is becoming an attractive way of treating infections in the UK. However, as well as having many advantages, an outpatient parenteral antibiotic therapy (OPAT) service potentially introduces new risks to staff and patients involved. In the United States, healthcare organizations are now prospectively analysing processes to try and prevent errors occurring using the Healthcare Failure Mode Effect Analysis (HFMEA) tool. The objectives of this study were to map out and agree the OPAT process and sub-processes and to identify potential OPAT system failures using steps 1-3 of the HFMEA tool, so that the resulting OPAT map can be used to design an OPAT service where risk is minimized. METHODS: The study was undertaken using a consensus development panel to which the HFMEA process was applied. Key stakeholders in the local OPAT process were invited to join the HFMEA team with the aim of describing and agreeing (defined as 100% participant agreement) an OPAT map, its sub-processes and potential OPAT system failures. RESULTS: The HFMEA team identified 6 processes, 67 sub-processes and 217 possible failures over the course of four meetings. Key areas identified in the OPAT map concerned identifying and checking patient suitability for an OPAT service, involvement of a multidisciplinary team and robust communication channels. CONCLUSIONS: An OPAT map was developed, which may serve as a practical model for other organizations setting up a similar service. PMID- 18408240 TI - Identification of differentially expressed genes in a Giardia lamblia WB C6 clone resistant to nitazoxanide and metronidazole. AB - OBJECTIVES: The characterization of differential gene expression in Giardia lamblia WB C6 strain C4 resistant to metronidazole and nitazoxanide using microarray technology and quantitative real-time PCR. METHODS: In a previous study, we created and characterized the G. lamblia WB C6 clone C4 resistant to nitazoxanide and metronidazole. In this study, using a microarray-based approach, we have identified open-reading frames (ORFs) that were differentially expressed in C4 when compared with its wild-type WB C6. Using quantitative real-time PCR, we have validated the expression patterns of some of those ORFs, focusing on chaperones such as heat-shock proteins in wild-type and C4 trophozoites. In order to induce an antigenic shift, trophozoites of both strains were subjected to a cycle of en- and excystation. Expression of selected genes and resistance to nitazoxanide and metronidazole were investigated after this cycle. RESULTS: Forty of a total of 9115 ORFs were found to be up-regulated and 46 to be down-regulated in C4 when compared with wild-type. After a cycle of en- and excystation, resistance of C4 to nitazoxanide and metronidazole was lost. Resistance formation and en-/excystation were correlated with changes in expression of ORFs encoding for major surface antigens such as the variant surface protein TSA417 or AS7 ('antigenic shift'). Moreover, expression patterns of the cytosolic heat-shock protein HSP70 B2, HSP40, and of the previously identified nitazoxanide-binding proteins nitroreductase and protein disulphide isomerase PDI4 were correlated with resistance and loss of resistance after en-/excystation. C4 trophozoites had a higher thermotolerance level than wild-type trophozoites. After en /excystation, this tolerance was lost. CONCLUSIONS: These results suggest that resistance formation in Giardia to nitazoxanide and metronidazole is correlated with altered expression of genes involved in stress response such as heat-shock proteins. PMID- 18408241 TI - Neuropsychological function and platelet monoamine oxidase activity levels in type I alcoholic patients. AB - AIMS: To explore neuropsychological function in two differentiated patterns of platelet monoamine oxidase B (MAO B) activity in alcoholic patients. METHODS: Neuropsychological examination and platelet MAO B activity extracted from blood were collected from 42 alcohol-dependent patients recruited in the alcoholism unit (NETER) of the Psychiatric Service of Santa Maria University Hospital. RESULTS: Alcoholics presented significantly low levels of platelet MAO B activity, when compared with control subjects; platelet MAO B activity in alcoholics classified as "under average subgroup" showed significant lower scores in the Raven Progressive Matrix and higher scores in hostility dimension, when compared with platelet MAO B activity in "above average subgroup." CONCLUSIONS: Results suggested platelet MAO B as a trait marker also to type I alcohol dependent patients and the two observed associations between platelet MAO B activity with neurocognitive measures of executive functions (nonverbal reasoning) and psychopathological dimension such as hostility may support the notion about the effect of platelet MAO B activity in the further development of an impulsive cognitive style. PMID- 18408242 TI - Habituation of bean (Phaseolus vulgaris) cell cultures to Quinclorac and analysis of the subsequent cell wall modifications. AB - BACKGROUND AND AIMS: The herbicide quinclorac has been reported to inhibit incorporation of glucose both into cellulose and other cell wall polysaccharides. However, further work has failed to detect any apparent effect of this herbicide on the synthesis of the wall. In order to elucidate whether quinclorac elicits the inhibition of cellulose biosynthesis directly, in this study bean cell calli were habituated to grow on lethal concentrations of the herbicide and the modifications in cell wall composition due to the habituation process were analysed. METHODS: Fourier transform infrared spectroscopy associated with multivariate analysis, cell wall fractionation techniques, biochemical analyses and the immunolocation of different cell wall components with specific monoclonal antibodies were used to characterize the cell walls of quinclorac-habituated cells. KEY RESULTS: Quinclorac-habituated cells were more irregularly shaped than non-habituated cells and they accumulated an extracellular material, which was more abundant as the level of habituation rose. Habituated cells did not show any decrease in cellulose content, but cell wall fractionation revealed that changes occurred in the distribution and post-depositional modifications of homogalacturonan and rhamnogalacturonan I during the habituation process. Therefore, since the action of quinclorac on the cell wall does not seem to be due to a direct inhibition of any cell wall component, it is suggested that the effect of quinclorac on the cell wall could be due to a side-effect of the herbicide. CONCLUSIONS: Long-term modifications of the cell wall caused by the habituation of bean cell cultures to quinclorac did not resemble those of bean cells habituated to the well-known cellulose biosynthesis inhibitors dichlobenil or isoxaben. Quinclorac does not seem to act primarily as an inhibitor of cellulose biosynthesis. PMID- 18408243 TI - Real and expected delivery rates of patients with myotonic dystrophy undergoing intracytoplasmic sperm injection and preimplantation genetic diagnosis. AB - BACKGROUND: This study aimed to analyse the reproductive outcome of a large cohort of myotonic dystrophy type 1 (DM1) patients undergoing ICSI and PGD. The secondary outcome parameter of this study was ovarian response as a way to express gonadal function in female DM1 patients. METHODS: Prospective cohort study. Real and expected cumulative delivery rates are descriptive. The reproductive outcome per cycle was compared with that of a control group of patients with X-linked recessive disorders. The comparative analysis of ovarian stimulation parameters in the study group versus the control group was carried out using both bivariate (crude) and multivariate (linear regression) analysis. RESULTS: Between 1995 and 2005, 205 cycles of ICSI and PGD were carried out for DM1 in 78 couples. The real cumulative delivery rate (max 6 cycles) overall was 46%. The expected overall cumulative delivery rate was 72%. Multivariate analysis did not show a significant difference in total dose of gonadotrophins used for ovarian stimulation between Group A (in which the female partner was affected) and a control group. CONCLUSIONS: This study shows that ICSI and PGD for DM1 offer good reproductive outcome, both in cumulative terms and per treatment cycle. There is no evidence of impaired gonadal function in female DM1 patients. PMID- 18408244 TI - Soluble lectin-like oxidized low density lipoprotein receptor-1 in type 2 diabetes mellitus. AB - The lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) can be proteolytically cleaved and released as soluble forms (sLOX-1). We have determined serums LOX-1 in type 2 diabetes and evaluated the effect of glucose and advanced glycation end products (AGEs) on sLOX-1 in vitro and in vivo. Endothelial cells were incubated with glucose or AGEs, and sLOX-1 in cell medium was measured. Serum sLOX-1 was measured in 219 diabetic patients and 187 controls by ELISA. The effect of lowering glucose and AGEs on sLOX-1 was determined in 38 poorly controlled diabetic patients after improvement in glycemic control. Incubation of endothelial cells with AGE-BSA led to a dose-dependent increase in sLOX-1, whereas the effect of glucose on sLOX-1 was less marked. Serum sLOX-1 was 9% higher in diabetic patients compared with controls (P<0.01). In the poorly controlled patients, serum sLOX-1 decreased by 12.5% after improvement in glycemic control (P<0.05). The magnitude of reduction in sLOX-1 correlated with the improvement in hemoglobin A1c and AGEs but not with the reduction in oxidized LDL. sLOX-1 level is increased in type 2 diabetes. Both glucose and AGEs are important determinants of LOX-1 expression, and lowering glucose and AGEs leads to a reduction in sLOX-1. PMID- 18408245 TI - Targeted quantitative analysis of Streptococcus pyogenes virulence factors by multiple reaction monitoring. AB - In many studies, particularly in the field of systems biology, it is essential that identical protein sets are precisely quantified in multiple samples such as those representing differentially perturbed cell states. The high degree of reproducibility required for such experiments has not been achieved by classical mass spectrometry-based proteomics methods. In this study we describe the implementation of a targeted quantitative approach by which predetermined protein sets are first identified and subsequently quantified at high sensitivity reliably in multiple samples. This approach consists of three steps. First, the proteome is extensively mapped out by multidimensional fractionation and tandem mass spectrometry, and the data generated are assembled in the PeptideAtlas database. Second, based on this proteome map, peptides uniquely identifying the proteins of interest, proteotypic peptides, are selected, and multiple reaction monitoring (MRM) transitions are established and validated by MS2 spectrum acquisition. This process of peptide selection, transition selection, and validation is supported by a suite of software tools, TIQAM (Targeted Identification for Quantitative Analysis by MRM), described in this study. Third, the selected target protein set is quantified in multiple samples by MRM. Applying this approach we were able to reliably quantify low abundance virulence factors from cultures of the human pathogen Streptococcus pyogenes exposed to increasing amounts of plasma. The resulting quantitative protein patterns enabled us to clearly define the subset of virulence proteins that is regulated upon plasma exposure. PMID- 18408246 TI - Bone mineral density in rheumatoid arthritis patients 1 year after adalimumab therapy: arrest of bone loss. AB - OBJECTIVE: To explore the effects of anti-tumour necrosis factor (TNF)alpha antibody therapy on bone mineral density (BMD) of the lumbar spine and femur neck in patients with rheumatoid arthritis (RA). METHODS: A total of 50 patients with active RA (DAS28> or =3.2) who started adalimumab (40 mg subcutaneously/2 weeks) were included in an open label prospective study. All patients used stable methotrexate and were allowed to use prednisone (< or =10 mg/day). The BMD of the lumbar spine and femur neck was measured before and 1 year after start of treatment. RESULTS: Disease activity at baseline (28-joint Disease Activity Score (DAS28)) and disease duration were inversely correlated with femoral neck BMD and lumbar spine BMD (p<0.05). Mean BMD of lumbar spine and femur neck remained unchanged after 1 year of adalimumab therapy (+0.3% and +0.3%, respectively). Of interest, a beneficial effect of prednisone on change in femur neck BMD was observed with a relative increase with prednisone use (+2.5%) compared to no concomitant prednisone use (-0.7%), (p = 0.015). CONCLUSION: In contrast to the progressive bone loss observed after conventional disease-modifying antirheumatic drug therapy, TNF blockade may result in an arrest of general bone loss. Consistent with previous observations, the data also suggest that the net effect of low-dose corticosteroids on BMD in RA may be beneficial, possibly resulting from their anti-inflammatory effects. PMID- 18408247 TI - Effect of intra-articular infliximab on synovial membrane pathology in a patient with a seronegative spondyloarthropathy. AB - OBJECTIVE: To demonstrate the efficacy of intra-articular infliximab in a patient with a persistent monarthritis who had previously had two arthroscopic synovectomies with limited success, and to determine the effect of intra articular infliximab on synovial membrane pathology METHOD: Arthroscopic synovial biopsy specimens were collected before and after treatment with intra-articular infliximab. The synovial tissue was stained for a range of inflammatory cell subsets, cell adhesion molecules and cytokines using immunohistochemical techniques and quantified using digital image analysis and a semiquantitative scoring method. RESULTS: Clinical improvement in the knee synovitis was seen after the first two intra-articular infliximab treatments, with a sustained clinical remission lasting for more than 12 months after the third treatment. Significant changes in cellular infiltration and expression of cytokines and cell adhesion molecules occurred as a result of treatment with intra-articular infliximab, with a reduction in some but not all cells in the inflammatory infiltrate, as well as a reduction in the expression of cell adhesion molecules (intercellular adhesion molecule-1 and vascular adhesion molecule-1) and production of cytokines (interleukin 1beta and tumour necrosis factor alpha). CONCLUSION: Intra-articular infliximab administration is a viable treatment for a persistent monarthritis resistant to other treatment options and can successfully modulate the inflammatory milieu within the synovial membrane. PMID- 18408248 TI - Change in cartilage morphometry: a sample of the progression cohort of the Osteoarthritis Initiative. AB - OBJECTIVE: The performance characteristics of hyaline articular cartilage measurement on magnetic resonance imaging (MRI) need to be accurately delineated before widespread application of this technology. Our objective was to assess the rate of natural disease progression of cartilage morphometry measures from baseline to 1 year in knees with osteoarthritis (OA) from a subset of participants from the Osteoarthritis Initiative (OAI). METHODS: Subjects included for this exploratory analysis are a subset of the approximately 4700 participants in the OAI Study. Bilateral radiographs and 3T MRI (Siemans Trio) of the knees and clinical data were obtained at baseline and annually in all participants. 160 subjects from the OAI Progression subcohort all of whom had both frequent symptoms and, in the same knee, radiographic OA based on a screening reading done at the OAI clinics were eligible for this exploratory analysis. One knee from each subject was selected for analysis. 150 participants were included. Using sagittal 3D DESSwe (double echo, steady-state sequence with water excitation) MR images from the baseline and 12 follow-up month visit, a segmentation algorithm was applied to the cartilage plates of the index knee to compute the cartilage volume, normalised cartilage volume (volume normalised to bone surface interface area), and percentage denuded area (total cartilage bone interface area denuded of cartilage). RESULTS: Summary statistics of the changes (absolute and percentage) from baseline at 1 year and the standardised response mean (SRM), ie, mean change divided by the SD change were calculated. On average the subjects were 60.9 years of age and obese, with a mean body mass index of 30.3 kg/m2. The SRMs for cartilage volume of various locations are: central medial tibia -0.096; central medial femur -0.394; and patella -0.198. The SRMs for normalised cartilage volume of the various locations are central medial tibia -0.044, central medial femur -0.338 and patella -0.193. The majority of participants had a denuded area at baseline in the central medial femur (62%) and central medial tibia (60%). In general, the SRMs were small. CONCLUSIONS: These descriptive results of cartilage morphometry and its change at the 1-year time point from the first substantive MRI data release from the OAI Progression subcohort indicate that the annualised rates of change are small with the central medial femur showing the greatest consistent change. PMID- 18408249 TI - Alcohol consumption is not protective for systemic lupus erythematosus. AB - OBJECTIVE: Several studies have suggested that alcohol drinking is protective for the development and progression of systemic lupus erythematosus (SLE). However, a protopathic bias might also explain this apparent association. Our objective was to investigate the association between alcohol consumption and incidence of SLE in a data set that has information on both current and pre-diagnostic alcohol consumption. METHODS: We performed an Internet-based case-control study of SLE. Cases were diagnosed within 5 years of the study and met > or =4 American College of Rheumatology criteria for SLE. The control participants were tightly matched to cases on demographic and socio-economic characteristics using a propensity score. Participants completed an online exposure assessment. We used conditional logistic regression analyses to test the association of current and pre diagnostic alcohol consumption with SLE. RESULTS: The sample comprised 114 cases with SLE and 228 matched controls. Current drinking (>2 days per week) was inversely associated with SLE (OR 0.35, 95% CI 0.13 to 0.98). Having more than two drinks per day was also inversely associated with SLE (OR 0.41, 95% CI 0.18 to 0.93). However, alcohol consumption before SLE diagnosis was not associated with the risk of SLE (p> or =0.4). Analysis of the change in drinking habits showed that people with lupus were more likely to quit drinking before (OR 2.25, 95% CI 0.96 to 5.28) or after (OR 2.38, 95% CI 0.88 to 6.49) being given the SLE diagnosis. CONCLUSIONS: Our results show that alcohol consumption before SLE diagnosis is not associated with the risk for SLE, and that individuals who develop SLE are more likely to quit. PMID- 18408250 TI - A single nucleotide polymorphism in the IRF5 promoter region is associated with susceptibility to rheumatoid arthritis in the Japanese population. AB - OBJECTIVES: Interferon regulatory factor 5 (IRF5) is a member of the IRF family of transcription factors, which regulate the production of proinflammatory cytokines. Polymorphisms in the IRF5 gene have been associated with susceptibility to systemic lupus erythaematosus (SLE) in Caucasian and Asian populations, but their involvement in other autoimmune diseases is still uncertain. Here, we assessed the genetic role of IRF5 in susceptibility to rheumatoid arthritis (RA) in Japanese subjects. METHODS: We selected 13 single nucleotide polymorphisms (SNPs) and a CGGGG insertion-deletion polymorphism in the IRF5 gene. We performed 2 sets of case-control comparisons using Japanese subjects (first set: 830 patients with RA and 658 controls; second set: 1112 patients with RA and 940 controls), and then performed a stratified analysis using human leukocyte antigen (HLA)-DRB1 shared epitope (SE) status. We genotyped the SNPs using TaqMan assays. RESULTS: A significant association of the rs729302 A allele with RA susceptibility was found in both sets (odds ratio (OR) 1.22, 95% CI 1.09 to 1.35, p<0.001 in the combined analysis). When the patients were stratified by the SE, the rs729302 A allele was found to confer increased risk to RA in patients that were SE negative (OR 1.50, 95% CI 1.17 to 1.92, p = 0.001) as compared with patients carrying the SE (OR 1.11, 95% CI 0.93 to 1.33, p = 0.24). In both sets, no genotyped polymorphisms were significantly associated with RA susceptibility, but rs729302 was significantly associated. CONCLUSIONS: These findings indicate that the promoter polymorphism of IRF5 is a genetic factor conferring predisposition to RA, and that it contributes considerably to disease pathogenesis in patients that were SE negative. PMID- 18408251 TI - Cartilage oligomeric matrix protein expression in systemic sclerosis reveals heterogeneity of dermal fibroblast responses to transforming growth factor beta. AB - OBJECTIVE: Cartilage oligomeric matrix protein (COMP) accumulates in systemic sclerosis (SSc) skin and is upregulated by transforming growth factor (TGF)beta. To further characterise the response to TGFbeta in SSc, we investigated TGFbeta1 and COMP expression and myofibroblast staining in SSc skin. METHODS: Skin biopsies from patients with diffuse cutaneous SSc (dSSc), limited cutaneous SSc (lSSc) and healthy controls were evaluated for COMP mRNA expression using real time PCR. COMP, alpha-smooth muscle actin (SMA) and TGFbeta were assessed in skin sections and in cultured fibroblasts by immunohistochemistry. Clinical disease status was assessed by the modified Rodnan skin score (mRSS). RESULTS: Myofibroblasts expressing SMA and COMP were found coexpressed in many cells in dSSc dermis, but each also stained distinct cells in the dermis. Cultured SSc dermal fibroblasts also showed heterogeneity for COMP and SMA expression, with cells expressing SMA, COMP, both or neither. TGFbeta treatment increased COMP and SMA-expressing cells. COMP mRNA expression in lesional skin from patients with dSSc correlated with the mRSS and TGFbeta1 staining. CONCLUSION: These findings suggest that TGFbeta upregulation of COMP and/or SMA expression in subpopulations of fibroblasts contributes to different pathways of fibrosis and that multiple TGFbeta regulated genes may serve as biomarkers for the degree of SSc skin involvement. PMID- 18408252 TI - Targeted drug delivery by in vivo coupling to endogenous albumin: an albumin binding prodrug of methotrexate (MTX) is better than MTX in the treatment of murine collagen-induced arthritis. AB - OBJECTIVE: To examine the effect of an albumin-binding prodrug of methotrexate (MTX) in the treatment of murine collagen-induced arthritis (CIA). METHODS: The prodrug AWO54 with the formula EMC-d-Ala-Phe-Lys-Lys-MTX binds selectively to the cysteine-34 position of endogenous albumin, which acts as a macromolecular drug carrier for MTX to the site of inflammation. The CIA model was used to evaluate the anti-arthritic effect of the compound after intravenous application. RESULTS: The albumin-bound form of AWO54 was efficiently cleaved by cathepsin B and plasmin, two proteases that are overexpressed in rheumatoid arthritis, and release a MTX lysine derivative. AWO54 suppressed CIA in a dose-dependent manner and was significantly better than MTX. To obtain a similar effect only about 20% of the MTX-equivalent dose of AWO54 had to be given. The efficacy of the drug was tested in two different stages of CIA: while both, MTX and AWO54 inhibited arthritis in an early stage of the disease, in a later stage only AWO54 showed a significant inhibitory effect in comparison with control. CONCLUSION: Targeted drug delivery by in vivo coupling of a prodrug of MTX to endogenous albumin is better than MTX in the treatment of CIA. PMID- 18408254 TI - Pancreatic contamination of mesenteric adipose tissue samples can be avoided by adjusted dissection procedures. AB - Mesenteric adipose tissue, located in the mesenterium of the intestines, is believed to play a central role in the development of obesity-related diseases. We have found that mesenteric fat samples harvested from rodents are frequently of poor quality, exhibiting partly degraded RNA. To investigate the background for this observation, we screened adipose tissue samples from two independent studies on rodents for markers of different tissues and cell types. We found that mesenteric adipose tissue samples of low quality are "contaminated" by pancreatic tissue. To locate the affected area, we dissected the mesenteric fat depots from 14 mice and measured abundance of pancreas-specific gene expression and amylase activity. As expected, we observed that the proximal section of the mesenterium, located near the pancreas, expressed pancreatic markers, whereas the distal sections did not. Approximately one-third of the mesenteric adipose tissue depots contained pancreatic tissue. Because the boundary between pancreas and mesenteric fat cannot be easily distinguished during dissection, we conclude that investigators should routinely exclude the proximal section of the mesenteric adipose tissue depot to avoid pancreatic contamination. PMID- 18408253 TI - Non-steroidal anti-inflammatory drug use does not appear to be associated with increased cardiovascular mortality in patients with inflammatory polyarthritis: results from a primary care based inception cohort of patients. AB - OBJECTIVES: There is controversy about the effects of non-steroidal anti inflammatory drugs (NSAIDs) on cardiovascular disease (CVD) mortality. The aim of this study was to explore associations between NSAID use and mortality in patients with inflammatory polyarthritis (IP). SUBJECTS AND METHODS: A total of 923 patients with new onset (IP), recruited to the UK Norfolk Arthritis Register (NOAR) between 1990-1994, were followed up to the end of 2004. Current medication was recorded annually for the first 6 years and then every 2-3 years. Rheumatoid factor (RF) and C-reactive protein (CRP) were measured. Logistic regression was used to calculate all cause and CVD mortality odds ratios (OR) for NSAID use at baseline and during follow-up, adjusting for gender and time-varying covariates: RF, CRP, joint counts, smoking, steroid use, DMARD use and other medication use. RESULTS: By 2004 there were 203 deaths, 85 due to CVD. At baseline, NSAIDs were used by 66% of patients. In final multivariate models, baseline NSAID use was inversely associated with all cause mortality (adjusted OR 0.62, 95% CI 0.45 to 0.84) and CVD mortality (adjusted OR 0.54, 95% CI 0.34 to 0.86). Interval NSAID use had weaker mortality associations: all cause mortality (adjusted OR 0.72, 95% CI 0.52 to 1.00), CVD mortality (adjusted hazard ratio (HR) 0.66, 95% CI 0.40 to 1.08). CONCLUSION: No excess CVD or all cause mortality was observed in NSAID users in this cohort of patients with IP. This is at variance with the literature relating to NSAID use in the general population. It is unclear whether this represents unmeasured confounders influencing a doctor's decision to avoid NSAIDs in the treatment of IP. PMID- 18408255 TI - Multi-scale finite element modeling allows the mechanics of amphibian neurulation to be elucidated. AB - The novel multi-scale computational approach introduced here makes possible a new means for testing hypotheses about the forces that drive specific morphogenetic movements. A 3D model based on this approach is used to investigate neurulation in the axolotl (Ambystoma mexicanum), a type of amphibian. The model is based on geometric data from 3D surface reconstructions of live embryos and from serial sections. Tissue properties are described by a system of cell-based constitutive equations, and parameters in the equations are determined from physical tests. The model includes the effects of Shroom-activated neural ridge reshaping and lamellipodium-driven convergent extension. A typical whole-embryo model consists of 10,239 elements and to run its 100 incremental time steps requires 2 days. The model shows that a normal phenotype does not result if lamellipodium forces are uniform across the width of the neural plate; but it can result if the lamellipodium forces decrease from a maximum value at the mid-sagittal plane to zero at the plate edge. Even the seemingly simple motions of neurulation are found to contain important features that would remain hidden, they were not studied using an advanced computational model. The present model operates in a setting where data are extremely sparse and an important outcome of the study is a better understanding of the role of computational models in such environments. PMID- 18408256 TI - Screw design alters the effects of stress relaxation on pullout. AB - Stress relaxation during pullout of a pedicle screw decreases the peak load and stiffness of the bone-screw interface. However, it is unknown whether this can be generalized to all types of screw designs. This study aimed to show whether screw design altered the effects of stress relaxation on the mechanical performance of the pedicle screw during pullout. Twelve calf vertebrae were obtained: six vertebrae were instrumented with 7.5x40 mm conical pedicle screws and the other six with 5.0x40 mm cylindrical pedicle screws. The screws with two different designs were pulled out using either a standard pullout or a stress relaxation pullout protocol. Both bone-screw interfaces had lower stiffness in the stress relaxation pullout model than in the standard pullout model, but it was significant in only the cylindrical design group (P<0.05). However, the stress relaxation and standard pullout models did not yield any difference in peak loads in either screw type. Although stress relaxation at the bone-screw interface can alter the mechanical performance of the screw, this may be eliminated by modifying the screw design. A better understanding of viscoelastic properties of the bone-screw interface may help improve implant design and thus, clinical outcomes. PMID- 18408257 TI - Diverse and composite features for ECG signals processing. AB - The automated diagnostic systems employing diverse and composite features for electrocardiogram (ECG) signals were analyzed and their accuracies were determined. Because of the importance of making the right decision, classification procedures classifying the ECG signals with high accuracy were investigated. The classification accuracies of multilayer perceptron neural network (MLPNN), recurrent neural network (RNN), and mixture of experts (ME) trained on composite features and modified mixture of experts (MME) trained on diverse features were compared. The inputs of these automated diagnostic systems were composed of diverse or composite features (wavelet coefficients and power levels of the power spectral density estimates obtained by the eigenvector methods) and were chosen according to the network structures. The conclusions of this study demonstrated that the MME trained on diverse features achieved accuracy rates which were higher than that of the other automated diagnostic systems trained on composite features. PMID- 18408258 TI - Soft layered concept in the design of metacarpophalangeal joint replacement implants. AB - The metacarpophalangeal (MCP) joint is crucial for hand function, but the joints are frequently affected by arthritis, leading to pain and disability. Joint replacement implants are used to replace the diseased MCP joint. This paper presents an investigation of applying the soft layered concept in the design of a new MCP joint replacement implant. Analytical methods were used to investigate the minimum film thickness for a novel MCP joint with a soft layer. The effect of load, entraining velocity, radial clearance, radius of the metacarpal head, elastic modulus and thickness of the soft layer were investigated. The soft layered joints show an enhanced predicted film thickness and some evidence of fluid film lubrication that should help to reduce wear rates. It may be beneficial for future MCP joint implant designs to utilise the soft layered joint concept. PMID- 18408259 TI - Thermal stability of acrylic bone cement powder under shelf storage conditions: an isothermal microcalorimetric study. AB - Heat-conduction isothermal microcalorimetry was used to measure the exothermic heat flow rate (Q) from the powder of a widely used commercially-available acrylic bone cement, Palacos R, when it interacted with ambient laboratory air, as a function of time, t, in the calorimeter, for up to 200 h. Four variants of the powder were used, these being unsterilized, sterilized using ethylene oxide gas, gamma-irradiated, in ambient air, with a minimum dosage of 2.5 Mrad, and gamma-irradiated, in ambient air, with a minimum dosage of 4.5 Mrad. In each case, the powder variant was tested after being stored on the shelf, under ambient conditions, for 2 days, 3 weeks and 9 months immediately following sterilization. Best-fit correlations between Q and t for each powder variant were determined. Then, this relationship was integrated over the period 14 h< or =t< or =200 h to give an estimate of the "effective" heat flow, Q(eff). For powder variants tested 2 days after being sterilized, the difference in their thermal stabilities (Qeff ranged from 0.19+/-0.01 to 0.62+/-0.03 microJ/g, respectively) was significant in the case of some pairs and not for others. However, for powders tested either 3 weeks or 9 months following sterilization, there was no significant difference between the means of Qeff (they ranged from 0.18+/-0.01 to 0.31+/-0.07 microJ/g) for any pair. These results suggest that an acrylic bone cement in which the powder is EtO-sterilized may be mixed with the liquid monomer for use in cemented arthroplasties after any length of time of shelf storage of the powder, under ambient conditions, whereas, for powders that are gamma irradiated and then stored under the same conditions, at least 3 weeks should elapse before they are used in these procedures. PMID- 18408260 TI - Medpor regulates osteoblast's microRNAs. AB - Porous polyethylene (PP or Medpor) is an alloplastic material worldwide used for craniofacial reconstruction. Although several clinical studies are available, there is a lack as regard the genetic effects. Because PP is always fixed on bone and the mechanism by which PP acts on osteoblasts is unknown, we therefore attempted to address this question by using microRNA microarray techniques to investigate the translation regulation in osteoblasts exposed to PP. The miRNA oligonucleotide microarray provides a novel method to carry out genome-wide microRNA profiling in human samples. By using miRNA microarrays containing 329 probe designed from Human miRNA sequence, we identified in osteoblast-like cells line (MG-63) cultured with Medpor (Porex Corporation, Fairburn, Georgia, USA) several miRNA which expression is significantly modified. We identified 16 up regulated miRNA (i.e. mir-337, mir-515-3p, mir-377, mir-153, mir-367, mir-152, let-7b, mir-92, mir-155, mir-424, mir-148b, mir-368, mir-18b, mir-520d, mir-20b, mir-128a) and 2 down-regulated miRNA (i.e. mir-143, mir-32). The data reported are, to our knowledge, the first study on translation regulation in osteoblasts exposed to PP. They can be relevant to better understand the molecular mechanism of bone regeneration and as a model for comparing other materials with similar clinical effects. PMID- 18408261 TI - The breath ammonia measurement of the hemodialysis with a QCM-NH3 sensor. AB - Recently, expired gases are analyzed non-invasively for monitoring the substances in the blood. Breath ammonia has been shown to correlate with BUN (blood urea nitrogen) and Cr (creatinine), both of which are indicators of solute removal in hemodialysis. In this study, breath ammonia concentration was continuously measured using a crystal oscillator QCM (quartz crystal microbalance) during the expiration of patients undergoing dialysis treatment. The results show that NH3 (ammonia) decreased gradually as the treatment proceeded. A strong correlation was observed between changes in the frequency of the QCM gas sensor and both the pre-dialysis BUN level (r=0.71, p<0.05) and the post-dialysis BUN level (r=0.90, p<0.05). NH3 was found to fall precipitously during dialysis. The differences were statistically significant. In addition, we found a statistically significant correlation between BUN and NH3 in expired gas. These results suggest that continuous measurement of NH3 is useful to assess the status of solute removal during hemodialysis. PMID- 18408262 TI - Fill it, shut it, and forget it...!!! PMID- 18408263 TI - Changing attitudes: from isolation to interaction. PMID- 18408264 TI - The effect of water purification systems on fluoride content of drinking water. AB - OBJECTIVE: The purpose of the present study was to determine the effect of different water purification systems on the fluoride content of drinking water and to compare the efficacy of these water purification systems in reducing the fluoride content. MATERIALS AND METHODS: Five different water purification systems were tested in this study. They were reverse osmosis, distillation, activated carbon, Reviva , and candle filter. The water samples in the study were of two types, viz, borewell water and tap water, these being commonly used by the people of Davangere City, Karnataka. The samples were collected before and after purification, and fluoride analysis was done using fluoride ion-specific electrode. RESULTS: The results showed that the systems based on reverse osmosis, viz, reverse osmosis system and Reviva showed maximum reduction in fluoride levels, the former proving to be more effective than the latter; followed by distillation and the activated carbon system, with the least reduction being brought about by candle filter. The amount of fluoride removed by the purification system varied between the system and from one source of water to the other. INTERPRETATION AND CONCLUSION: Considering the beneficial effects of fluoride on caries prevention; when drinking water is subjected to water purification systems that reduce fluoride significantly below the optimal level, fluoride supplementation may be necessary. The efficacy of systems based on reverse osmosis in reducing the fluoride content of water indicates their potential for use as defluoridation devices. PMID- 18408266 TI - Comparison of allele frequency for HLA-DR and HLA-DQ between patients with ECC and caries-free children. AB - BACKGROUND: Early childhood caries (ECC) is one of the most common diseases of childhood. The etiology of ECC is multifactorial and both genetic and environmental factors play important roles in the pathogenesis of the disease. Genetic variations in the hosts may contribute to changes in the risk for dental caries. Genetic factors such as human leukocyte antigen (HLA) have recently been suggested as a predisposing factor. AIM: The aim of this study was to look for an association between HLA-DRB1 and HLA-DQB1 with ECC for developing new strategies for the diagnosis as well as the prevention of the disease. DESIGN: In this study, we extracted the genomic DNAs from whole blood samples of 44 patients with ECC and 35 caries-free children by the salting-out method. We amplified the genomic DNA by PCR-SSP and then HLA-typing was performed for all alleles. RESULTS: The results revealed a significant increase in the frequency of HLA DRB1*04 in the patient group (P=0.019). The odds ratio for this allele was detected to be 10. The frequency of HLA-DQB1 alleles was not significantly different between the two groups. CONCLUSION: The above results suggest that HLA DRB1*04 is associated with the susceptibility to ECC. Thus HLA-DRB1*04 detection as a molecular marker for early diagnosis of ECC may be recommended. PMID- 18408265 TI - Effect of oil pulling on Streptococcus mutans count in plaque and saliva using Dentocult SM Strip mutans test: a randomized, controlled, triple-blind study. AB - BACKGROUND: Oil pulling has been used extensively for many years, without scientific evidence or proof, as a traditional Indian folk remedy to prevent teeth decay, oral malodor, bleeding gums, dryness of throat and cracked lips, and for strengthening the teeth, gums, and jaws. AIMS: The aim of this study was to evaluate the effect of oil pulling with sesame oil on the count of Streptococcus mutans in plaque and saliva of children, using the Dentocult SM Strip mutans test, and to compare its efficacy with that of chlorhexidine mouthwash. MATERIALS AND METHODS: Twenty age-matched adolescent boys were selected based on information obtained through a questionnaire. They were divided randomly into two groups: the control or chlorhexidine group (group I) and the study or oil pulling group (group II); there were ten subjects in each group. Plaque and saliva samples were collected from all the 20 subjects on the strips from the Dentocult SM kit and, after incubation, the presence of S. mutans was evaluated using the manufacturers' chart. The study group practiced oil pulling with sesame oil and the control group used chlorhexidine mouthwash for 10 min every day in the morning before brushing. Samples were collected from both groups after 24 h, 48 h, 1 week, and 2 weeks and the efficacy of oil pulling was compared with that of chlorhexidine mouthwash. RESULTS: There was a reduction in the S. mutans count in the plaque and saliva samples of both the study and the control groups. The reduction in the S. mutans count in the plaque of the study group was statistically significant after 1 and 2 weeks (P=0.01 and P=0.008, respectively); the control group showed significant reduction at all the four time points (P=0.01, P=0.04, P=0.005, and P=0.005, respectively, at 24 h, 48 h, 1 week, and 2 weeks). In the saliva samples, significant reduction in S. mutans count was seen in the control group at 48 h, 1 week, and 2 weeks (P=0.02, P=0.02, P=0.008, respectively). CONCLUSION: Oil pulling can be used as an effective preventive adjunct in maintaining and improving oral health. PMID- 18408267 TI - Witkop's tooth and nail syndrome: a multifaceted approach to dental management. AB - Witkop's tooth and nail syndrome is a rare autosomal dominant disorder of ectodermal dysplasia characterized by hypodontia and nail dysplasia. Mutations in MSX-1 have been shown to be associated with this syndrome. There is failure of development and eruption of the dentition. Tooth shape may vary; the most common forms are conical and narrow crowns. The nails may be spoon shaped and slow growing and affect both finger and toe nails. The nail involvement is more severe in childhood. The present case describes a 14-year-old boy who showed the characteristic features of Witkop's syndrome. A multifaceted approach to the dental management of the patient is discussed. PMID- 18408268 TI - Management of inverted impacted primary incisors: an unusual case. AB - The prevalence of impacted primary teeth has been reported to be very rare and it has been reported to occur in 1:10,000 times, most commonly involving the primary second molars. This report discusses the presence of an impacted inverted maxillary primary incisors and their management by surgical intervention in a 5 year old boy. The inverted primary maxillary central incisors were extracted and prosthetic rehabilitation was done to improve the esthetics, speech and psychosocial behaviour of the child. Co-incidentally the permanent maxillary incisors were also found to be hypoplastic although the parents did not give any history of trauma to the maxillofacial complex. It is important for pediatric dental surgeons to carefully monitor the eruption of primary and permanent teeth for early identification and management of developmental anomalies. PMID- 18408269 TI - Orthodontic uprighting of impacted mandibular permanent second molar: a case report. AB - The mandibular second molars can become impacted beneath the crown of the first molars due to various causes and fail to erupt normally. Presented herewith is a case report of orthodontic uprighting of a mesioangular impacted mandibular right permanent second molar. Though various treatment options were available, an uprighting push spring appliance was used as it is easy to fabricate and produces distal tipping and uprighting of the impacted tooth without the necessity of surgical assistance, bone removal, or splinting. The uprighting of the mandibular second molar was achieved within two months. PMID- 18408270 TI - Greenstick fracture of the mandible: a case report. AB - This case report is an insight in to pediatric traumatology whereby bilateral greenstick fracture of condyle is used as a means to discuss the incidence and anatomic considerations for the management of the same, highlighting the fact that dental surgeons require a unique understanding of the anatomy, growth considerations, healing pattern and operative management involving minimal manipulation while managing pediatric facial fractures. PMID- 18408271 TI - Disappeared roots: a case report. AB - Tooth resorption is a perplexing problem for all dental practitioners. The etiologic factors are vague, the diagnoses are educated guesses and, often, the chosen treatment does not prevent the rapid disappearance of the calcified dental tissues. This becomes all the more confusing if the tooth in question is a pulpally involved young permanent tooth. Presented in this report is the case of an upper first young permanent molar that underwent complete root resorption following root canal therapy and obturation. PMID- 18408273 TI - Salt and hypertension in man. PMID- 18408272 TI - Cleidocranial dysplasia: a case report. AB - Cleidocranial dysplasias is an autosomal dominant disorder that presents with skeletal dysplasia. The dental manifestations are mainly delayed exfoliation of primary teeth and delayed eruption of permanent teeth, with multiple impacted supernumeraries. This report addresses the complex nature of the treatment modalities. In our patient, surgical exposure of unerupted teeth was done with orthodontic traction. Post-surgical follow-up was uneventful. PMID- 18408274 TI - Acquired cystic disease of kidney in chronic renal failure in jordan. AB - Acquired cystic disease of the Kidney (ACDK) is a well documented complication of end stage renal failure in patients managed conservatively or supported by dialysis. The aim of the present study is to highlight the scope of the problem among our patients, and to evaluate the clinical characteristics of the disease. A Total of 124 patients with chronic renal failure (106 on dialysis, 18 non dialyzed) were studied with ultrasonography by the same radiologist. ACDK was defined as the presence of four or more cysts in each kidney. The cysts should have sharp smooth wall with acoustic enhancement. In non-dialyzed patients, 5.5% had ACDK, 33.5% had one to three cysts and 61% had no cysts. Among dialysis patients 42.5% had ACDK, 26% had one to three cysts and 31.1% had no cysts. During of dialysis therapy and age were greater in patients with ACDK (74 +/- 30 months. 58 +/- 3 years respectively) compared to those with no cysts (11 +/- 8 months. 26 +/- 4 years). The major clinical complications were intracystic hemorrhage in three cases and stones in seven cases. No correlation was noted between the presence of cysts and the original kidney disease, gender, hematocrit or adequacy of dialysis. We conclude that, ACDK occurs commonly in patients with chronic renal failure; however, patients are generally older and have been on dialysis longer than patients without cysts. Ultrasound, in the hands of an experienced radiologist, is a relatively adequate screening tool for ACDK and its complications. PMID- 18408275 TI - Doppler ultrasound evaluation of hemodialysis vascular access. AB - Brescia-Cimino arteriovenous fistulas (AVF) ad synthetic grafts are the usual forms of vascular accesses for hemodialysis. Although angiography has been the traditional means of imaging these vascular systems, colour Doppler flow imaging (CDFI) offers a non-invasive method of evaluating AVF dysfunction. We retrospectively evaluated 22 patients with clinical evidence of access dysfunction who were studied by CDFI and of whom 14 also underwent angiography. We analyzed the results of whom the 14 patients who had both CDFI and angiography as angiography was impossible in the remaining eight patients due to difficulty with cannulation. Eight patient had thrombosis on CDFI and angiopraphy in all eight patients confirmed these findings. CDFI showed six stenoses, all of which were proven on angiopraphy. Overall,, CDFI correctly identified all lesions that were seen angiographically ginging a sensitivity and specifificity of 100%. Moreover, CDFI detected two cases of pseudoaneurysms which were missed by angiography. CDFI provides an adequate means of evaluating AVF dysfunction and should be the initial imaging technique of choice. PMID- 18408276 TI - Peritonitis in Patients on CAPD at King Khalid University Hospital: Less Infection-rate with More Center-experience. AB - Continuous ambulatory peritoneal dialysis (CAPD) was started at the King Khalid University Hospital in 1986. Peritonitis remains the most significant complication of the procedure. Earlier rates of peritonitis were high, but after gaining sufficient experience, the rates are declining. To evaluate the change in peritonitis trend, 55 new patients who were recruited to our CAPD program between the periods October 1993-October 1996 were analyzed for the development of peritonitis on annual basis. There were 37 (67%) males and 18 (33%) females with a mean age of 43.3 years (range 12-72 years). A total of 34 episodes of peritonitis were recorded with a rate of 1.5 episode/patients year in 1993, 0.5 episode/patient year in 1994 and 0.8 in 1995. only 40% of episodes showed positive cultures whereas 60% remained culture-negative despite use of recommended modified culture techniques. Organisms causing peritonitis included staphylococcus eipdermides (6.7%), E. coli (3.3%), Streptococcus fecalis (3.3%) and pseudomonas (6.7%). Out of 34 episodes of peritonitis, 29 (85.3%) showed response to treatment and five episodes could only be treated after removal of catheter. Of the 29 episodes that responded to treatment, three relapsed and one had recurrent infection . However, all were successfully treated though one responded only after removal of catheter. Thus, a total of six catheters (20%) necessitated removal and replacement. In spite of high diabetic patients population in our series (27.2%) only one died of peritonitis related sepsis and another died of myocardial infarction after clearing the infection. Thus mortality remains low in spite of potential risk. Although we still use straight system CAPD rather than Y system peritonitis rates have declined considerable and we hope that the procedure will gain more acceptability amongst patients with ESRD in Saudi Arabia. PMID- 18408277 TI - Acute renal failure in newborn: etiology and mortality rate in jordan patients. AB - A retrospective multicenter study of 38 cases of acute renal failure (ARF) in newborns was carried out from April 1992 to September 1995 in the pediatric department so f a group hospitals of he Royal Medical Services in Jordan, to evaluate the etiology and the mortality rate in the first month of life. ARF was diagnosed according to the urine out put, serum level of creatinine and blood urea nitrogen. Neonatal asphyxia, the most common cause in this study, accounted for 42% (N=16) of renal failure and was associated with the highest mortality rate 70% (N=11). Drugs (aminoglycosides or vancomycin), the second most common cause of renal failure, accounted for 14% (N=7), but no patient sided. Septicemia accounted for 15.7% (N=6) of renal failure; one patients died. Three cases were due to genitorreal anomalies, one died with real agenesis. The other six cases were of different causes, four of them died. None of the newborns was treated by dialysis. We conclude that neonatal asphyxia is the most common cause of ARF in our series, and it carries poor prognosis. However, drug induced ARF has relatively good prognosis. The overall prognosis of ARF in the newborn was rather poor, the cumulative mortality, without dialysis, being of ARF in the newborn was rather poor, the cumulative mortality, without dialysis, being 45% in this study. PMID- 18408278 TI - Renal artery stenosis in renal transplantation presentation and management. AB - A review of the angioplasty records between 1990 and 1995 at the King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia revealed ten cases of transplant renal artery stenosis (RAS). The diagnosis in these cases was confirmed by renal angiography and all were treated by angioplasty. All study patients presented with uncontrolled hypertension in spite of multiple medications; eight had renal functional impairment and two patients had recurrent unexplained pulmonary edema in addition. Six patients had undergone end-to-end anastomosis, while four had end-to-side anastomosis of the artery during transplantation. Four had cadaveric renal transplants and six had living donor renal transplants. Eight of these patients responded well to angioplasty with marked improvement in their renal function and reduction in the number of anti hypertensive medications. In one patient, it was not possible to pass the catheter through the stenosis and the patient underwent surgical reconstruction, while in another patient there were multiple stenotic lesions involving the external iliac and the transplant renal arteries suggesting atherosclerotic changes. We conclude that renal artery stenosis should be suspected in patients after renal transplant if they have uncontrolled or worsening hypertension, unexplained renal impairment or presentation with unexplained recurrent pulmonary edema. Renal angiography should be considered as part of the investigation of hypertension in renal transplant patients, and if the RAS is confirmed, angioplasty should be the procedure of choice. PMID- 18408279 TI - Clinical approach to ophthalmoplegia in renal transplant recipients differential diagnosis and case illustration. AB - A 57 year-old man presented six months after undergoing renal transplantation with the orbital apex syndrome comprising of retro-orbital pain, visual loss and complete ophthalmoplegia. Pathological examination from the granulomatous-like mass in the apex of the right orbit showed branching hyphae with the characteristic appearance of aspergillus. The differential diagnosis of such a lesion in an immunocompromised patient is discussed. PMID- 18408280 TI - Imperforate hymen with bilateral hydronephrosis in a neonate. AB - Imperforate hymen is a rare diagnosis in the neonatal period. Bilateral hydronephrosis and renal failure are rare complications of an imperforate hymen. We present the case of a newborn with undiagnosed imperforate hymen complicated with renal failure. PMID- 18408281 TI - Renal replacement therapy in qatar. PMID- 18408282 TI - Change or evolution. PMID- 18408283 TI - Genetically transmitted renal diseases in children: a saudi perspective. AB - A significant number of renal diseases in children occur as a result of genetic susceptibility. It is believed that such diseases, particularly those with an autosomal recessive transmission, may be more common in places with a high rate of consanguineous marriages as in the Middle East including the Kingdom of Saudi Arabia (KSA). Although a comparison of the published literature from the KSA with that from other countries outside the region support such a possibility, too few studies have been published from the region to definitely answer the question. Preliminary observations indicate that the children in the Kingdom probably have a higher incidence of polycystic kidney disease, familial juvenile nephronophthisis, congenital urological anomalies and familial nephrotic syndrome. It is reasonable to speculate that whereas some diseases may be common, others may be rare because of a low gene frequency in the local population. PMID- 18408284 TI - Adequacy of Hemodialysis: It is not Only Urea we Have to Monitor. PMID- 18408285 TI - Incidence and types of malignant tumors in renal transplant recipients: a single center experience. AB - We reviewed the incidence and types of de novo malignancies which developed in 792 renal transplant recipients who received their grafts between 1979 and 1996 and followed up for an average period of 5.6 years. There were 56 malignant tumors detected in 54 patients, representing an overall incidence of 6.8%. These tumors occurred in a relatively young group of patients whose average age was 33.4 years at the time of transplantation and 40 years at the time of diagnosis of malignancy. The average latency period between transplantation and malignant disease was 7.4 years for solid tumors and 16 months for Kaposi's sarcoma. Tumors included 39 Kaposi's sarcomas, four malignant lymphomas, three hepatomas, two bladder cancers, one renal cell carcinoma of the allograft, two colorectal cancers, two thyroid cancers, one adenocarcinoma of unknown primary, one nasopharyngeal carcinoma and one leiomyosarcoma of the uterus. Kaposi's sarcoma was the most common malignancy and comprised 70% of all tumors in this study. It is imperative for renal transplant recipients to have diligent follow-up at regular intervals for early detection of cancer. PMID- 18408286 TI - Survey of the current status of renal transplantation in the arab countries. AB - To evaluate the current status of renal transplantation in the Arab world, a questionnaire was mailed to prominent renal transplant physicians and surgeons in the Arab countries to get information related to number, donor type, immunosuppressive therapy used and survival of graft and patients who received renal transplantation. Furthermore, data about rehabilitation and legislation in the Arab countries were also included. The cost of renal transplantation was estimated from historical data. There were replies from 14 out of 22 (70%) countries with a total population of approximately 210 million (84%). Renal transplantation started in most of these countries in the late seventies and early eighties. In all but one, the renal transplant programs started with live related donations, and continued over the past 1 5-20 years with this type of donation. There have been 5,680 such operations, performed in a growing number of transplant centers. Legislation organizing donation from cadaveric donors has been established in only seven of the fourteen countries (50%) and, to date, there are only five countries with specialized centers to organize organ donation. The overall average graft survival is 87%, 80%, 60%, while patient survival is 96%, 85%, 70%, during one, five and ten years respectively. Full rehabilitation averaged 50%, partial rehabilitation averaged 40% and complete dependence averaged 10% after renal transplantation in the Arab countries. We conclude that there is still a need for more efforts to increase the number of renal transplantation, passing legislations that organize organ donation, and increase utilization of cadavers in renal transplantation in the Arab countries. PMID- 18408287 TI - Spectrum of renal osteodystrophy in dialysis patients at a tertiary hospital, riyadh, saudi arabia. AB - To evaluate renal osteodystrophy (ROD), bone biopsies were performed in 57 patients with end-stage renal failure (ESRF) on dialysis, 46 on hemodialysis (HD) and 11 on peritoneal dialysis (PD). There were 29 males (mean age of 42 years) and 28 females (mean age of 39 years). Relevant presenting clinical features were pruritus in 46 cases, bone pains in 32, acute pseudogout in three, bone deformities in two, conjunctiva! calcification in two, cutaneous calcification in two, and corneal calcification in one. The mean value of predialysis blood investigations were as follows: urea 33.9 mmol/L, creatinine 913 umol/L, bicarbonate 18 mmol/L, calcium 2.36 mmol/L, albumin 40 g/L, phosphorus 1.69 mmol/L, alkaline phosphatase 178 U/L, parathyroid hormone 543 pmol/L, magnesium 1.06 mmol/L and aluminum 1.81 mmol/L. Skeletal survey showed no changes in 24 patients (42%), hyperparathyroid cystic changes of bones in seven, osteoporosis as the predominant features in seven, mixed picture of ROD in 12, subperiosteal resorption of the metacarpals in two, osteosclerosis (Rugger Jersey Spine) in two and osteomalacia in two patients. Bone mineral density (BMD) was measured by dual x-ray absorptiometry in the lumbar spine (LS) and femoral neck (FN). All patients had low BMD (both LS and FN). Bone biopsy (BBX) revealed mixed picture in 30 cases, predominantly secondary hyperparathyroid changes in 10, mild hyperparathyroid changes in five, predominant osteoporosis in three, osteomalacia in four, aplastic (adynamic) bone in four, and aluminum deposition in one. All of the patients who showed evidence of bone involvement on BBX had abnormal BMD suggesting that BMD is a good non-invasive screening test for ROD but indiscriminative for the type of bone disease. BBX still remains the diagnostic tool to differentiate and classify different types of bone disease. PMID- 18408289 TI - Post-dialysis Solutes Rebound: Comparison of Two Protocols for Hemodialysis. AB - The blood urea concentration is artificially low immediately following high efficiency dialysis. Post-dialysis urea rebound correlates with hemodialysis efficiency, and is inversely correlated with dialysis treatment time. We evaluated the effect of variation in the length of hemodialysis treatment on urea, creatinine and other solutes rebound. We used two protocols for hemodialysis, using 500-ml dialysate/min and similar dialyzed blood volume. Protocol A: hemodialysis with blood flow of 300 ml/min for four hours; protocol B: hemodialysis with blood flow of 400 ml/min for three hours. Fifteen stable anuric patients with end- stage renal disease (ESRD) were hemodialysed using each protocol, three sessions a week, for a period of two weeks. The mean dialyzed blood volume in protocol A and B was 66 +/- 4 and 67 +/- 4.8 liters/session, respectively. The mean blood flow in protocol A was 286 +/- 22 ml/min, and in protocol B was 395.3 +/- 13 ml/min. The mean urea pre and immediately post dialysis in patient using protocol A was 20.5 +/- 5.4 and 5.55 +/- 2.2 mmol/L, respectively. While in protocol B it was 19.8 +/- 4.6 and 6 +/- 1.68 mmol/L. The mean urea, one hour post dialysis, in protocol A was 6.51 +/- 1.9 Vs 8.04 +/- 2.6 mmol/L in protocol B (P value < 0.003). The percent rebound of mean blood urea concentration in protocol A and B one hour post dialysis was 17.3% vs 34%, respectively. Predialysis creatinine in protocol A and B was 894 +/- 156.8 vs 907.9 +/- 163 umol/L, respectively (P= 0.4). The immediately post-dialysis creatinine in protocol A and B, was 317+/- 98.4 vs 331 +/- 72.0 micromol/L (P = 0.4), while one hour later it was 398.6 +/- 104.0 vs 442.6 +/- 107.2 micromol/L, respectively (P value < 0.007). The percent rebound of creatinine was 25.6% in protocol A vs 33.5% in protocol B. These results show significant difference between the two protocols, and confirm increased rebound of urea and creatinine one hour post dialysis with shorter time of dialysis treatment. PMID- 18408288 TI - Radionuclide assessment of patients with urinary tract infection and vesico ureteric reflux using tc-99m-dimercaptosuccinic Acid. AB - The purpose of this study was to evaluate pediatric patients with urinary tract infection and to identify the risk of developing renal scarring and its relationship to the presence of vesico-ureteric reflux. The subjects in this study were 29 pediatric patients between two months and 11 years of age with a history of urinary tract infection among which 69% were females. All the patients underwent radio-isotope scanning using Technetium-99-m-dimercaptosuccinic acid, 4 6 weeks after the eradication of the acute infection. Micturating cysto urethrography and renal ultrasound examinations were performed before the radio isotope scanning. Isotope scanning showed evidence of scarring in 34% of the kidneys, while ultrasound scanning was positive in only 29% of the kidneys. The agreement between the results of the two examinations was in 88% of the kidneys. Reflux was noted in 53% of the kidneys of which 62% showed scarring by isotope scanning. Only 11% of the kidneys with no reflux showed evidence of scarring, while 92% of kidneys with severe reflux had renal scarring. In patients with reflux, 57% of the kidneys in female patients and 50% of the kidneys in male patients developed renal scarring evident on DMSA scanning. Absence of reflux had a high predictive value for absence of renal scarring, while the presence of reflux did not necessarily imply the presence of renal scarring but the possibility increased with the severity of reflux. PMID- 18408290 TI - Treatment of Post-transplant Erythrocytosis with Enalapril. AB - This prospective study was carried out to estimate the efficacy and safety of Enalapril therapy in post-transplant erythrocytosis. Thirteen long-term renal allograft recipients (11 males and two females) with increased hematocrit values (>51%) and elevated red cell mass were treated with Enalapril for 12 weeks; their age ranged from 18 to 54 years. At the end of the study period the mean hematocrit values decreased from 53.9% to 46.7%, red cell mass significantly decreased from 50.8 ml/kg to 40.5 ml/kg. During the following six months without Enalapril treatment, an increase in hematocrit was observed reaching a mean of 47.5%. In conclusion, enalapril can be safely and efficiently used to treat post transplant erythrocytosis. PMID- 18408291 TI - Paroxysmal nocturnal hemoglobinuria and renal failure. AB - In this report we present two cases of acute renal failure in paroxysmal nocturnal hemoglobinuria (PNH) patients, who both eventually recovered their renal function. We also highlight some of the possible pathophysiological changes, which may contribute to renal failure in patients with PNH. PMID- 18408292 TI - Chyluria associated with renal vasculitis. AB - Chyluria is usually associated with abnormal retrograde or collateral flow of lymph from intestinal lymphatics into lymphatics of the kidney, ureter or bladder. Chyluria has been described in patients with lymphoma, carcinoma, trauma, abscess, tuberculosis, filariasis, pregnancy and stenoses of the thoracic duct. In this case report we describe a patient who presented with chyluria associated with severe hyperlipidemia due to nephrotic syndrome induced by microscopic polyangiitis. Laboratory, histological and radiological examination did not show evidence of filarial infestation, tuberculous infection or malformation of the lymphatic system. Treatment with corticosteroids and cyclophosphamide resulted in improvement of hyperlipidemia and disappearance of chyluria. PMID- 18408293 TI - Extraction of urinary stone. PMID- 18408294 TI - Acquired cystic renal disease and hemodialysis. PMID- 18408295 TI - Pathogenesis of acute renal failure: shock-kidneys. AB - Causes of acute renal failure (ARF) are summarized. The article focuses on "shock kidneys" as they occur following traumatic or septic shock. There may be low grade intermittent but persisting endotoxemia in the former together with other factors like rhabdomyolysis, and marked endotoxemia at least for a few hours in the latter. Endotoxin is a prime cause of release of noxious cytokines like tumor necrosis factor-alpha (TNFa). At present, many studies support the evidence for its role in multi-organ failure (MOF). One can account for endotoxemia along with bacterial translocation through the gastrointestinal mucosa if there is transient mesenteric ischemia during shock. Hence, monocyte-macrophages can be stimulated to release their cytokines that predispose to MOF. The cell biology of renal tubular changes in ARF is then briefly discussed in order to mention new therapeutic approaches. PMID- 18408297 TI - Acute renal failure in the tropics. PMID- 18408296 TI - Prevention of acute renal failure. PMID- 18408298 TI - Acute renal failure in pregnancy. PMID- 18408299 TI - Acute renal failure in the intensive care unit. PMID- 18408300 TI - Rhabdomyolysis and Myoglobin-induced Acute Renal Failure. PMID- 18408301 TI - Acute renal failure due to snake-bite: clinical aspects. PMID- 18408302 TI - Acute Renal Failure and HELLP Syndrome: A Single Center's Experience. AB - A total of 52 patients were referred to our center from gynecology and obstetric units in our area with acute renal failure during the last two years. Seven patients were found to have so called syndrome of hemolysis (H), elevated liver enzymes (EL) and low platelets (LP) associated with acute renal failure. The syndrome can easily be confused with other diagnoses like hemolytic uremic syndrome, idiopathic thrombotic thrombocytopenic purpura and disseminated intravascular hemolysis. Six patients had renal biopsies to confirm the diagnosis, while one did not consent for biopsy. Four patients were found to have acute tubular necrosis, one had acute cortical necrosis and one was not enough for interpretation. We conclude that the patients can easily be misdiagnosed if we are not familiar with the diagnosis and that the overall prognosis is good if the patient survives the acute stage. PMID- 18408303 TI - Slow continuous ultrafiltration with dialysis in patients with acute renal failure in the intensive care unit. PMID- 18408304 TI - Acute renal failure: six months pilot study in qatar. AB - Over a period of six months, 55 patients out of 11,216 (0.49%) admitted to the hospital developed acute renal failure (ARF). The diagnosis of ARF was based on the usual criteria, a sudden rise in blood urea nitrogen and creatinine with or without oliguria. Patients age ranged between 15 and 81 years with a mean of 51.9 years. Renal ischemia (69%) and nephrotoxic drugs (16.3%) were the two main etiologic factors. Among the causes of ischemia, septic shock was the commonest (29%), followed by severe hypotension due to several causes such as hemorrhage, burns, severe diarrhea and cardiogenic shock (25.4%), and ACE inhibitors (10.9%). ARF was associated with an average of 15.8 days stay in hospital versus 5.1 days for the overall hospital admissions. Immediate management of hypotension by intravenous fluid replacement, vasopressor agents and the necessary surgical intervention was appropriately considered. Intravenous frusemide was used for oliguric patients. Intermittent hemodialysis was used in 18 patients and continuous venovenous hemofiltration in six patients. Twelve patients with ARF due to ischemia died, while there were no deaths in the nephrotoxic group (p < 0.05). The overall mortality was (21.8%), which had no correlation with patient age. All non-oliguric patients survived with the mortality being exclusively in the oliguric group. PMID- 18408305 TI - Acute renal failure in jordan. AB - We evaluated 215 patients with acute renal failure (ARF) in three centers in Jordan over an 18 months period. Their ages ranged between 12-90 years, and 120 of them were males. Parenchymal renal insult was the commonest cause of ARF as it was seen in 125 patients (58%). Pre-renal azotemia was seen in 60 patients (28%) and acute obstructive uropathy in 30 patients (14%). At presentation, 152 patients (70.7%) were oligo-anuric, while 63 (29.3%) were non-oliguric. Forty patients (18.6%) required dialysis support; 30 of them were in the renal failure group (75%). Thirty-two of the 40 patients were oliguric-aruric. Complete recovery of renal function was achieved in 80% for the whole group, and in 64% of those with parenchymal renal insult. Forty-seven patients (21.9%) died; 35 of them (63.9%) were in the renal group, and 37 patients (78%) were oligo-anuric. Sepsis and cardiac complications were together responsible for almost 75% of the deaths. PMID- 18408307 TI - Acute renal failure in Sudan. PMID- 18408306 TI - Opinion Survey about the Evaluation and Management of Acute Renal Failure in Saudi Arabia. AB - To evaluate the approach of physicians to the diagnosis and management of acute renal failure (ARF) in the Kingdom of Saudi Arabia, a questionnaire was mailed to nephrologists, physicians attending to renal failure patients, specialists working in intensive care unit (ICU) and the general physicians in 110 hospitals, which have either an ICU or a dialysis unit. The questions were related to the areas of evaluation, conservative management, dialysis therapy, and prognosis of ARF. There were 135 responses from 76 hospitals (69%); 37 of small size (< 150 beds), 21 of medium size (151-400 beds), and 18 of large size (401-1200 beds). There were 69 respondents from the small hospitals, 34 from the medium-sized, and 32 from the large hospitals. According to the respondents, the most encountered ARF patients were in the intensive care units and were most likely due to sepsis and nephrotoxic drugs. There were no differences among the respondents in the areas of initial evaluation, conservative and/or emergency treatment. However, the nephrologists were significantly more willing to follow-up their ARF patients and to carry more specific diagnostic procedures (i.e., renal biopsy) and specific advanced therapeutic procedures (i.e., dialysis), More ARF patients are being treated by continuous renal replacement therapy (CRRT) than intermittent hemodialysis or peritoneal dialysis. The minority of the respondents believed that the prognosis of ARF had not improved much, despite the improvement in diagnosis and therapy. They attributed this to the change in the demographics of ARF, since more ICU and elderly patients are seen in practice. We conclude that nephrologists are indispensable for the management of acute renal failure. More efforts may be needed to recruit nephrologists to hospitals in Saudi Arabia. A local fellowship program may help in this regard. Furthermore, CRRT facilities, at least in the large hospitals, are required. PMID- 18408308 TI - Bilingual medical journal. PMID- 18408309 TI - Acute renal failure in the neonate. PMID- 18408310 TI - Osteodystrophy in chronic renal failure patients. PMID- 18408311 TI - Hypertension in pregnancy: presentation, management and outcome - a retrospective analysis of 135 cases. AB - In view of continuous controversies regarding hypertension in pregnancy and lack of reports on this subject in Saudi Arabia, we retrospectively studied 135 episodes of hypertension in pregnancy in 123 patients over a 9-years period. Fifty-one (37.8%) were primiparas, the remaining were multiparas from 2 nd up to 14 11 pregnancy. Of the multiparas, 44% had a history of pregnancy induced hypertension, 26% of chronic hypertension, and 13% of diabetes mellitus. " HELLP" syndrome occurred in nine and eclampsia in four pregnancies. There were 10 foetal deaths, eight of them presented on admission. Chronic hypertension and diabetes mellitus approached significance as risk factors for foetal death (logistic regression analysis). For the whole group, the time of delivery was the only factor with significant impact on the Apgar score (multiple regression analysis), with an additional negative influence of diabetes mellitus and intravenous hydralazine in the multiparas. When the patients with chronic hypertension and diabetes mellitus were not included, hydralazine lost its significance. The finding suggests that intravenous hydralazine should be given in multiparas with caution, particularly in the presence of chronic hypertension and diabetes mellitus. There was a significant relation of the birth weight to the time of delivery, the serum levels of total protein and, especially in multiparas, of uric acid, confirming the importance of these factors as markers of the severity of pregnancy induced hypertension. The hypertension persisted in patients not known to have chronic hypertension in 9.6% of pregnancies during a follow-up ranging from 2 months up to 9 years. PMID- 18408312 TI - Incidence of treated end-stage renal disease in asir region, southern saudi arabia. AB - Incidence of treated end-stage renal disease (t-ESRD) in Saudi Arabia is not well documented and only few reports are available. This study was conducted to determine the incidence of t-ESRD in Asir region. The study period included January 1995 to December 1995. All new cases admitted for chronic dialysis treatment in all MOH-hospitals with a diagnosis of ESRD were included. There were 114 Saudi and 10 non-Saudi Patients. Among the 114 Saudi patients, there were 64 (56.1%) males and 50 (43.9%) females. Mean age 47.86 +/- 19.86 years (range 9-90 years). The overall incidence of t-ESRD was 214.9 pmp. Age adjusted incidence for males and females were 243 pmp and 186 pmp, respectively. There were no differences between males and females (X 2 =2.019, p = < 0.05). The majority of the patients (41.2%) did not have definitive diagnosis. Among the known causes, glomerulonephritis was diagnosed or suspected in 56.7%, diabetes was the cause of ESRD in 16.4% of the cases. This is the highest incidence of ESRD ever reported in Saudi Arabia. Geographical, environmental, genetical factors may have been contributing factors. PMID- 18408313 TI - Outcome of congenital hydronephrosis in qatar. AB - Three hundred and eleven patients diagnosed to have congenital hydronephrosis proven by postnatal ultrasonography in a ten-year period between 1987 and 1996 were studied retrospectively. These were classified into mild degree of hydronephrosis in H (64%), moderate in 94 (30%) and severe in 18 (6%) patients, according to first post natal sonography and depending on the measurement of the anterio-posterior transverse diameter of the pelvi-calyceal system. Significant obstructive uropathy was documented in 66 patients by Technetium-Diethylene triamine penta acetic acid renogram (Tc99m DTPA Renogram Vesicoureteric reflux was found in 22 patients by voiding cysto-urethrogram. On follow-up the hydronephrosis regressed spontaneously in 99 (31.8%), was stable in 167 (53.6%) and needed operative intervention in 45 (14.4%) patients. The types of surgical operations were pyeloplasty in 49% and ureteric re-implantation in 27%. Fifteen (83%) of the children with severe hydronephrosis and 32 (34%) of the moderate hydronephrosis group were operated o while none of the children with the mild form required operative intervention. We recommend ultrasound follow-up for mild hydronephrosis and a coordinated approach and work-up for congenital moderate and severe hydronephrosis. Surgery should only be considered if renal function is affected. PMID- 18408314 TI - Neurological disorders in renal transplant recipients. AB - The charts of 175 renal transplant recipients were retrospectively reviewed. The mean duration of follow up since the transplantation was 4.17 +/- 1.66 years. Apart from cyclosporin induced tremor, which occurred almost in all patients, 22 patients (12.5%) had neurological disorders during their follow up (mean annual incidence of 3%). Eight patients had epileptic seizures, seven had strokes, four had neurological infections, two developed a pseudotumor cerebri syndrome, two neuropathies, one myopathy and one conus medullaris infarction. This study demonstrates that neurological disorders are not uncommon in renal transplant recipients and that their mechanisms are variable and may be related to the underlying diseases such as hypertension and diabetes; to the operation itself; to the side effects of immunosuppression agents or rarely, they can accompany graft rejection. PMID- 18408315 TI - Fasting During the First Year of Transplantation: Is it Safe? AB - During the month of Ramadan 1417H we studied 14 patients all transplanted with live related kidneys. All the patients were transplanted between one and seven months before the month of Ramadan. All the patients were on azathioprine and prednisolone and the mean plasma creatinine level was 95 +/- 15 micromol/L. They all completed fasting the month of Ramadan successfully. Medical evaluation showed that there was no clinical adverse effect. There was no significant change in serum values of creatinine, calcium, protein, sodium, potassium, chloride, HC03, or liver function tests. There was, however, a significant elevation of total plasma proteins, urea, uric acid, hemoglobin, triglycerides, cholesterol, urinary sodium and potassium, concentration (p=< 0.05). Our study shows that fasting in Ramadan during the first year of transplantation is possible, nevertheless further studies with large number of patients are necessary to confirm this specially in patients receiving cyclosporine. PMID- 18408316 TI - Understanding immunoglobulin IgA nephropathy. PMID- 18408317 TI - Sarcoidosis with partial reversibility of renal failure: two case reports with review of literature. AB - Sarcoidosis is one of the granulomatus disorders affecting many organ systems of the body. Renal impairment in sarcoidosis is rare and occurs usually as a result of long standing hypercalcemia or hypercalciuria with nephrocalcinosis or renal stones. Sarcoid glomerulopathy and tubulo-interstitial granulomatus involvement have been described. We report two cases of sarcoidosis, the first with interstitial nephritis and anterior uveitis without evidence of granuloma. The patient was normocalcemic and normocalciuric. The second case presented with nephritic range proteinuria and severe renal insufficiency with a history of recurrent parotid swelling seven years before diagnosis. Renal biopsy showed non caseating granulomas in the tubulo-interstitial region. Both patients showed good response to steroid therapy, however, there is still residual renal insufficiency six months after therapy. In conclusion, renal sarcoidosis although a rare presentation, should be considered in the presence of extra-renal manifestation of sarcoidosis as it is amenable to treatment. PMID- 18408318 TI - Renal replacement therapy in bahrain. PMID- 18408319 TI - Renal replacement therapy in sultanate of oman. PMID- 18408320 TI - Salt excretion and hypertension. PMID- 18408321 TI - Acquired renal cysts: the other side of the coin. PMID- 18408322 TI - Radionuclide assessment of patients with urinary tract infection and vesico ureteral reflux using tc--99m-dimercaptosuccinic Acid. PMID- 18408323 TI - Anthelmintic macrolactams from Nonomuraea turkmeniaca MA7381. AB - A new macrolactam, fluvirucin B0 (1), and two known macrolactams, Sch 38516/fluvirucin B1 (2) and Sch 39185/fluvirucin B3 (3), have been isolated from an acetone extract of a strain of Nonomuraea turkmeniaca. These compounds were isolated by bioassay-guided fractionation as part of our search for new anthelmintics. The structures of these compounds were elucidated by comparison of their NMR and MS data to those of previously reported fluvirucins, and confirmed by 2D-NMR. 1approximately 3 exhibited in vitro activity (EC90 <1.0 approximately 1.7 microg/ml) against Haemonchus contortus larvae, but were ineffective in reducing worm counts in vivo against Heligmosomoides polygyrus in mice at 50 mg/kg dosed intramuscularly. PMID- 18408324 TI - Biosynthesis of 3'-deoxy-carbamoylkanamycin C in a Streptomyces tenebrarius mutant strain by tacB gene disruption. AB - Streptomyces tenebrarius H6 mainly produces three kinds of antibiotics: apramycin, carbamoyltobramycin and some carbamoylkanamycin B. In our present study, a dehydrogenase gene tacB in the tobramycin biosynthetic gene cluster was disrupted by in-frame deletion. The result of TLC bio-autograph analysis demonstrated the disruption mutant strain produced apramycin and a new antibiotic. The new antibiotic was identified as 3'-deoxy-carbamoylkanamycin C by MS and NMR analysis after isolation and purification. The disruption mutant was restored to produce carbamoyltobramycin in a complementation experiment by the intact tacB gene. Our studies suggested that the tacB gene encodes a 6' dehydrogenase, which reduces the 6'-hydroxyl group of paromamine to a keto group, thus facilitating the transfer of an aminogroup to form neamine. This study is the first report on the generation of a tobramycin derivative by gene engineering, and will contribute to clarify the complete biosynthetic pathway of tobramycin. PMID- 18408325 TI - Structural determination and proposed biosynthesis of alcanivorone, a novel alpha pyrone produced by Alcanivorax jadensis. AB - A novel alpha-pyrone designated as alcanivorone was found in a culture broth of the marine bacterium, Alcanivorax jadensis, and its structure was determined by an analysis of 1D NMR, 2D NMR and MS data. Alcanivorone was produced by A. jadensis only when sodium pyruvate was added to the culture medium as a carbon source. Incorporation experiments using stable isotope-labeled pyruvate indicated that alcanivorone was biosynthesized from four molecules of pyruvate. PMID- 18408326 TI - Studies on terpenoids produced by actinomycetes. 5-dimethylallylindole-3 carboxylic Acid and A80915G-8"-acid produced by marine-derived Streptomyces sp. MS239. AB - As a result of screening for terpenoids produced by marine-derived Streptomyces sp. MS239, two new terpenoids named 5-dimethylallylindole-3-carboxylic acid and A80915G-8''-acid were isolated and their structures were determined mainly by NMR analyses. PMID- 18408328 TI - Scabronine J, a new cyathane-type diterpenoid from the basidiomycete Sarcodon scabrosus. AB - A new cyathane-type diterpenoid, named scabronine J was isolated from the fruiting bodies of the mushroom Sarcodon scabrosus. The structure of scabronine J was elucidated on the basis of spectroscopic studies including 2D NMR experiments. PMID- 18408327 TI - Chrysogenamide A from an endophytic fungus associated with Cistanche deserticola and its neuroprotective effect on SH-SY5Y cells. AB - Chrysogenamide A (1), a new member of the macfortine group of alkaloids, along with four known compounds (2 approximately 5) were identified from Penicillium chrysogenum No. 005, an endophytic fungus associated with Cistanche deserticola Y. C. Ma. The new structure was elucidated on the basis of comprehensive spectral analysis. 1 exhibited a neurocyte protection effect against oxidative stress induced cell death in SH-SY5Y cells. PMID- 18408329 TI - Chemical synthesis and characterization of J46 peptide, an atypical class IIa bacteriocin from Lactococcus lactis subsp. cremoris J46 Strain. AB - Bacteriocin J46 is a 27-residue polypeptide produced by Lactococcus lactis subsp. cremoris J46 in fermented milk. The natural form of J46 (nJ46) exhibits a broad antimicrobial spectrum. Herein, we produced the synthetic form of J46 (sJ46) by solid-phase chemical synthesis. The biochemical and physico-chemical properties of sJ46, as well as its antimicrobial activity, were found to be identical to those of its natural counterpart nJ46. It showed a potent antimicrobial activity against both lactic acid bacteria and other Gram-positive microorganisms. (1)H NMR conformational analysis of sJ46 indicates that it adopts a flexible random coil structure. PMID- 18408330 TI - Fungal metabolite, epoxyquinol B, crosslinks proteins by epoxy-thiol conjugation. AB - Epoxyquinol B (EPQB) is a fungal metabolite, containing two alpha,beta-epoxy ketones. We previously showed that EPQB inhibited the signal transduction involved in angiogenesis through the binding to cysteine residues of receptor kinases. However, the inhibitory mechanism was undefined. In this report, we found that one EPQB molecule is covalently bound to two L-cysteine molecules through two epoxide residues on EPQB. Furthermore, EPQB crosslinked binding proteins through the cysteine residues. These results suggest that EPQB inhibits receptor kinases by crosslinking with other protein or by intramolecular crosslinking. PMID- 18408332 TI - [Regulation of allergic responses by Kampo medicines]. PMID- 18408331 TI - Structure-activity relationship of pamamycins: effect of side chain length on aerial mycelium-inducing activity. AB - Two pamamycin homologues with different side chain lengths were isolated from Streptomyces sp. HKI-0118. Aerial mycelium-inducing activity decreased by ca. 1/10 per methylene unit in the side chain. PMID- 18408333 TI - [Possible effects of ingredients in foods on brain function]. PMID- 18408334 TI - [Effect of biotin ingestion on the improvement of hypertension in SHRSP]. PMID- 18408335 TI - [Clinical application of dietary supplements]. PMID- 18408336 TI - [Diverse physiological and pharmacological functions of fatty acids]. PMID- 18408337 TI - [Measurement of endothelial function in humans]. PMID- 18408338 TI - [Recent and potential drug candidates for treatment of Parkinson's disease]. PMID- 18408339 TI - [The development of therapeutic agents for Parkinson's disease and problems in their clinical application]. PMID- 18408340 TI - [Drug-induced cutaneous toxicity]. PMID- 18408341 TI - [Pharmacological and clinical profiles of long-lasting erythropoiesis-stimulating agent (darbepoetin alfa; NESP]. PMID- 18408342 TI - [The novel bowel cleansing agent Visiclear (Visiclear Tablets)]. PMID- 18408343 TI - Simple flow-through solubility measurement apparatus and its effectiveness for hazard assessment of particles/fibers. PMID- 18408344 TI - A European framework to address psychosocial hazards. AB - Over the past decades, emphasis has been placed on the changing nature of work and new forms of risk that could negatively affect employee health and safety. These are mainly associated with new types of occupational hazards that have been termed psychosocial. Issues such as work-related stress, bullying and harassment are now receiving attention on a global basis and efforts have been made to address them at the workplace level. However, it has been acknowledged that despite developments of policy in this area, there still appear to be a broad science-policy gap and an even broader one between policy and practice. The WHO Network of Collaborating Centers in Occupational Health has, since the late 1990s, been supporting a dedicated program of work on psychosocial factors and work-related stress. Part of the Network's work is currently focusing on the translation of existing knowledge into practice in the area of psychosocial risk management. This program has identified that the optimum way forward lies in the development of a European framework for psychosocial risk management. This framework will serve as the basis for coordination of research activities and preventive action with an emphasis on evidence based interventions and best practice on an international basis. PMID- 18408345 TI - Effect of nursing assistance tools on preventing musculoskeletal pain among staff in schools for disabled children. AB - Objective is to clarify whether nursing assistance tools (a mat with attached handles, a pair of trousers with knee pads and a waist holding belt) prevent musculoskeletal pain, such as low back pain and upper arm pain, and depression, and improve the burden on the lower back and upper arm among staff in schools for disabled children. This study design was a non-randomized intervention trial. The subjects were 41 staff in two schools for disabled children in Japan. Nursing assistance tools were used with the intervention group to help with their nursing activities. We investigated the one-month prevalence of low back pain and the degree of burden on the lower back using a questionnaire at the baseline and at the end point 4 to 6 months later. The prevalence of low back pain did not change significantly in either group. In the intervention group, the prevalence of upper arm pain decreased from 47.6% at the baseline to 23.8% at the end point (p=0.063). The percentage of participants with a high level of burden on the lower back from excretory nursing activity decreased from 57.1% at the baseline to 33.3% at the end point (p=0.063) in the intervention group. These results suggest that nursing assistance tools may prevent upper arm pain and improve the burden on the lower back among staff in schools for disabled children; however, these tools did not significantly prevent low back pain and depression. PMID- 18408346 TI - Association of occupational stress and social support with health-related behaviors among chinese offshore oil workers. AB - To explore the relationship of occupational stress and social support with health related behaviors of smoking, alcohol usage and physical inactivity, a cross sectional survey was conducted among 561 offshore oil installation workers of a Chinese state-owned oil company. They were investigated with a self-administered questionnaire about socio-demographic characteristics, occupational stress, social support and health-related behaviors. Logistic regression analysis was used to study the association between occupational stress, social support and health-related behaviors and adjusted for age, educational level, marital status, duration of offshore work and job title. Of 561 workers, 218 (38.9%) were current smokers, 124 (22.1%) current drinkers, and 354 (63.1%) physically inactive in their leisure time. Further multivariate logistic regression analysis indicated that: (1) Current smoking was significantly negatively related with perceived stress from "Safety" (OR=0.74; 95% CI=0.58-0.94) and lack of supervisors' instrumental support (OR=0.34; 95% CI=0.18-0.65); (2) Current drinking was significantly positively related to perceived stress from "Interface between job and family/social life" (OR=1.32; 95% CI=1.02-1.70) and "Organizational structure" (OR=1.35; 95% CI=1.06-1.74), but was significantly negatively related to poor emotional support from friends (OR=0.54; 95% CI=0.62-0.96); (3) Physical inactivity after work was significantly positively associated with perceived stress from "Safety" (OR=1.44; 95% CI=1.16-1.79) and lack of instrumental support from both supervisors (OR=1.74; 95% CI=1.16-2.65) and friends (OR=1.68; 95% CI=1.06-2.42). The findings suggest that psychosocial factors of occupational stress and social support at offshore oil work might affect workers' health related behaviors in different ways. PMID- 18408347 TI - Stability in urine of authentic phenylglyoxylic and mandelic acids as urinary markers of occupational exposure to styrene. AB - Phenylglyoxylic acid (PhGA) and mandelic acid (MA) are two popular urinary markers of occupational exposure to styrene, but PhGA has been considered to be relatively unstable when urine samples are stored. This study was initiated to examine the stability of PhGA in urine under two storage conditions, i.e., at room temperature (at 25 degrees C) and in refrigeration (at 4 degrees C) for up to 14 days. The experiments showed that no substantial decrease was observed in either PhGA or MA even at room temperature within one day, but, depending on urine samples, a gradual decrease in PhGA took place both at 4 degrees C and more markedly at 25 degrees C when kept for a week. Further reduction was observed in two weeks even at 4 degrees C. No reduction was observed in MA up to two weeks both at 4 degrees C and at 25 degrees C. The observation on stability of MA and limited stability of PhGA (i.e., no significant decrease for 4 days when stored as refrigerated) was confirmed by a repeated experiment. Further analyses disclosed that PhGA when stored at 4 degrees C tended to be more unstable when urine samples were alkaline (e.g., at pH 8) rather than acidic (e.g., at pH 6 or below), but the trend varied subject to individual samples. Thus, the practical recommendation is that urine samples should be analyzed on the day of collection if all possible, or kept at 4 degrees C, or more preferably at -20 degrees C. Refrigeration will allow storage of up to 4 days without substantial decrease in either PhGA or MA. PMID- 18408348 TI - Urinary excretion of an oxidative stress marker, 8-hydroxyguanine (8-OH-Gua), among nickel-cadmium battery workers. AB - The relationship between oxidative stress and carcinogenic metals including nickel and cadmium is a matter of interest. To assess the oxidative stress status of workers exposed to nickel and cadmium simultaneously, we determined urinary excretion of 8-hydroxyguanine (8-OH-Gua), a urinary oxidative stress marker. Our subjects were 66 (64 males and 2 females) nickel-cadmium battery workers. Spot urine and blood samples were collected. The levels of cadmium in blood (Cd-B) and nickel in urine (Ni-U) were determined by graphite furnace atomic absorption spectrophotometry. 8-OH-Gua in urine was analyzed using a high performance liquid chromatography-electrochemical detector (HPLC-ECD) system. Data on age, sex, duration of present work and smoking status were also obtained from each subject. Creatinine-adjusted 8-OH-Gua was significantly correlated with age, Ni-U and Cd-B in univariate analysis, while multivariate analysis revealed that Ni-U and Cd-B were significant independent variables and that these two biological exposure indices were positively correlated with 8-OH-Gua. The data were also analyzed in the context of mixture toxicity. The subjects were divided into groups based on median level of Ni-U and Cd-B (2.86 mug/g creatinine and 0.23 mug/dl, respectively). Workers with high Ni-U/high Cd-B (Group IV) had the highest levels of 8-OH-Gua levels (GM (GSD), 21.7(2.0)), followed by those with high Ni-U/low Cd B (11.5(1.6) Group III), those with low Ni-U/high Cd-B (8.9(1.9) Group II), and those with low Ni-U/low Cd-B (8.5(1.5) Group I). The p values of Students' t tests between Group I and Group II, III and IV were 0.847, 0.050 and <0.001, respectively. The combined effect of Cd and Ni on the urinary excretion of 8-OH Gua departed from additivity. PMID- 18408349 TI - Evaluation of the effects of shift work on nutrient intake: a cross-sectional study. AB - The aim of this study was to investigate the effects of shift work on nutrient intakes, including macronutrient intake and micronutrient intake, in a large sample size. The study population included 2,254 male manual workers, 20-59 yr of age, employed in a factory. A self-administered diet history questionnaire was used. The subjects were classified into 3 groups according to their work schedule: (i) fixed day workers; (ii) shift workers without midnight shifts; and (iii) shift workers with midnight shifts. The nutrient intakes of the groups were compared by age group. There was a significant difference in nutrient intakes between subjects aged 20-29 yr and those aged 30 yr and over. Among subjects aged 20-29 yr, the energy density value for saturated fat and the energy adjusted intakes of calcium, potassium, vitamin A, and vitamin B1 were the lowest among shift workers with midnight shifts. Among subjects aged 30 yr and over, the total energy intake was the highest among shift workers with midnight shifts; the difference was significant compared to fixed day workers. Shift workers with midnight shifts had the highest intake of cereals among subjects 30 yr of age and older. In conclusion, there were no significant differences in nutrient intakes between fixed day workers and shift workers without midnight shift. Shift work, particularly midnight shift work, affected nutrient intake. The impact of shift work on nutrient intakes differed by age and the type of shift work. PMID- 18408350 TI - Prevalence and determinants of workplace violence of health care workers in a psychiatric hospital in Taiwan. AB - Workplace violence, a possible cause of job stress, has recently become an important concern in occupational health. This study determined the prevalence of workplace violence and its risk factors for employees at a psychiatric hospital in Taiwan. A questionnaire developed by ILO/ICN/WHO/PSI was first translated and validated. It was then used to survey the prevalence of workplace violence in the last 12 months experienced by all nursing aides, nurses, and clerks at the hospital. Multiple logistic regression models were constructed to discover the determinants of violence. A total of 222 out of 231 surveyed workers completed a valid questionnaire. The one-year prevalence rates of physical violence (PV), verbal abuse, bullying/mobbing, sexual harassment, and racial harassment were 35.1, 50.9, 15.8, 9.5, and 4.5%, respectively. The prevalence of PV at this hospital was higher than that reported by other countries for the health sector. A high anxiety level was associated with the occurrence of PV. These results need to be corroborated by future investigation. A training program may be required for high risk groups to reduce workplace violence. PMID- 18408351 TI - Fertility after artificial insemination using a soybean-based semen extender in sheep. AB - The present study aimed to compare the fertility of ewes intrauterinally inseminated with frozen-thawed semen using a soybean-based semen extender (AndroMed) with those of ewes intrauterinally inseminated with frozen-thawed semen using a Tris-based extender containing either egg yolk or BSA. Suffolk ewes (n=104) were treated with an intravaginal sponge containing 40 mg fluoroprogesterone acetate (FGA) for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin to induce estrus and ovulation during the non-breeding season (July, 2007). Intrauterine insemination was carried out 40-46 h after removal of the FGA sponge (n=90), regardless of the incidence of estrus. The pregnancy rates were not significantly different among the semen extenders containing egg yolk (64.5%) or BSA (58.6%) and AndroMed extender (56.7%). The lambing rates (64.5, 55.2 and 56.7% for the semen extenders containing egg yolk, BSA and AndroMed, respectively) and prolificacy (1.59 to 1.75) were also not significantly different. The present results indicate that an egg yolk-containing semen extender can be replaced with the non-animal derived extender AndroMed, which could be used for intrauterine insemination using frozen-thawed ram semen without reducing fertility. PMID- 18408352 TI - Defined system for in vitro production of porcine embryos using a single basic medium. AB - We have previously indicated that porcine blastocysts can be produced by in vitro fertilization (IVF) and culture (IVC) in chemically defined porcine gamete medium (PGM) and porcine zygote medium (PZM)-5, respectively, In the present study, the effects of basic media and macromolecular components on in vitro maturation (IVM) were investigated to develop a defined system for in vitro embryo production using a single basic medium through IVM, IVF and IVC. Porcine immature oocytes were matured in porcine oocyte medium (POM) or modified North Carolina State University (mNCSU) 37, which were supplemented with either 10% (v/v) porcine follicular fluid (pFF) or 3 mg/ml polyvinyl alcohol (PVA) as a macromolecular component (designated POM+pFF, POM+PVA, mNCSU37+pFF and mNCSU37+PVA). In the maturation with mNCSU37+PVA, the percentages of oocytes that reached the metaphase II stages were significantly lower than those in the other treatments. Following IVM with the above media, oocytes were treated with an electrical stimulus and cycloheximide for parthenogenetic activation and were cultured in PZM-5 for 5 days. The rates of cleavage and blastocyst formation of parthenogenetic oocytes were significantly lowered for maturation with mNCSU37+PVA compared with the other treatments, while there were no significant differences in the total numbers of cells in blastocysts among the treatments. Following IVF and IVC, the rates of penetration, male pronucleus formation, cleavage and blastocyst formation were significantly lower when oocytes were matured in mNCSU37+PVA than in other maturation media. The normal fertilization rate was significantly higher in POM+PVA compared with the other treatments, although the total number of cells in blastocysts was reduced with the addition of PVA to both POM and mNCSU37 compared with pFF supplementation. These results demonstrate that porcine blastocysts can be produced by the defined system using a single basic medium. PMID- 18408353 TI - Inheritance of histone H3 methylation in reprogramming of somatic nuclei following nuclear transfer. AB - Successful cloning requires reprogramming of epigenetic information of the somatic nucleus to an embryonic state. However, the molecular mechanisms regarding epigenetic reprogramming of the somatic chromatin are unclear. Herein, we transferred NIH3T3 cell nuclei into enucleated mouse oocytes and evaluated the histone H3 dimethyl-lysine 4 (H3K4me2) dynamics by immunocytochemistry. A low level of H3K4me2 in the somatic chromatin was maintained in pseudo-pronuclei. Unlike in vitro fertilized (IVF) embryos, the methylation level of nuclear transfer (NT) embryos was significantly increased at the 8-cell stage. NT embryos showed lower H3K4me2 intensity than IVF embryos at the 2-cell stage, which is when the mouse embryonic genome is activated. Moreover, the H3K4me2 signal was weak in the recloned embryos derived from single blastomeres of the NT embryos, whereas it was intense in those from IVF embryos. Two imprinted genes, U2afbp-rs and Xist, were abnormally transcribed in cloned embryos compared with IVF embryos, and this was partly correlated to the H3K4me2 level. Our results suggest that abnormal reprogramming of epigenetic markers such as histone acetylation and methylation may lead to dysregualtion of gene expression in cloned embryos. PMID- 18408354 TI - Affected homologous chromosome pairing and phosphorylation of testis specific histone, H2AX, in male meiosis under FKBP6 deficiency. AB - A gene for FK506 binding protein 6 (Fkbp6) expresses during a specific stage of male and female meiosis. Disruption of the gene influences male reproduction, i.e. arrests spermatogenesis, but not female reproduction. Using the mouse model (targeted disruption), the role of the gene in homologous chromosome pairing has been demonstrated in a previous study. For further understanding the function of Fkbp6 in chromosome synapsis, we evaluated chromosome pairings during male meiosis in the as/as rat, a spontaneous null mutation, and compared them with those of the mouse model. Electron microscopy of the pachytene nuclei unveiled several types of abnormal chromosome pairing in the rat model, as shown in the mouse previously. The frequencies of aberrant pairings in the knockout mice and mutant rats were 42 of 67 nuclei (62.7%) and 20 out of 74 nuclei (27.0%), respectively. In order to clarify the mechanism of male specific infertility in Fkbp6 deficiency, the localization of gammaH2AX, a marker protein of XY chromosome inactivation during male meiosis, was examined. Immunostaining of gammaH2AX unveiled normal localization of the molecule to XY chromosomes (XY body) in both models, showing the independency of FKBP6 in sex chromosome inactivation. Besides the XY body, focal localization of gammaH2AX was observed in accordance with the unsynapsed chromosomes in both types of null animal. These results indicate the fundamental role of Fkbp6 in homologous chromosome synapsis during male meiosis. In conclusion, male specific infertility under Fkbp6 deficiency remains unsolved. PMID- 18408355 TI - Cloning and sequence analysis of multiple splice variants of lactate dehydrogenase C in yak testes. AB - Lactate dehydrogenase C (LDH-C) has been reported to play a role in the energy metabolism of mammal spermatozoa. However, the functions and expression patterns of LDH-C still remain unclear. In order to elucidate the functions and expression patterns of LDH-C, we cloned the cDNA of yak LDH-C. Total RNA was extracted from yak testes and reverse transcribed and amplified by PCR. The full length open reading frame (ORF) of LDH-C and its five splice variants were obtained. The full length ORF contained 999 bp encoding a 332-amino-acid protein that showed 100% identity with bovine LDH-C. Compared with the full length ORF of LDH-C, the five variants used the same start codon as the full length ORF and encoded 5 putative proteins. In detail, variants 1 (missing the coding sequence of exon 6 and 7) and 2 (missing the coding sequence of exon 7) bear the entire nicotinamide-adenine dinucleotide (NAD) binding domain and an active site. Variants 3 (missing the first 42 nuleotides of exon 4) and 4 (missing the coding sequences of exons 5, 6 and 7) lack part of the NAD binding domain but contained the entire active site. Variant 5 (missing the coding sequence of exons 4 and 7) lacks a large part of the NAD binding domain and the entire active site. Native polyacrylamide gel electrophoresis was performed to determine if the splice variants can be translated into proteins. However, native PAGE detected no specific bands from yak testes and bovine spermatozoa. This study suggests the alternative splicing of LDH-C is ubiquitous in bovine testes and might be involved in regulation of LDH-C expression. The findings also help to elucidate the functions of LDH-C. PMID- 18408356 TI - Guidelines for management of idiopathic normal pressure hydrocephalus. AB - With the rapid aging of Japanese society, medical care of the elderly has become an important social issue. Among various disorders manifesting dementia, gait disturbance, and urinary incontinence in the elderly population, normal pressure hydrocephalus (NPH), especially of idiopathic type (iNPH), is becoming noteworthy. The Guidelines for management of iNPH in Japan are created in compliance with the evidence-based medicine methods and published in 2004. This English version is made to show the diagnosis and treatment of iNPH with reference to the socio-medical background in Japan and to promote the international research on iNPH. They propose three diagnostic levels; possible, probable, and definite. They indicate the diagnostic importance of high convexity tightness and dilated sylvian fissure with mild to moderate ventriculomegaly on coronal magnetic resonance imaging. The cerebrospinal fluid tap test is regarded as an important diagnostic test because of its simplicity to perform and high predictability of the shunt efficacy. The use of programmable valves at shunt surgeries is recommended. Flowcharts for diagnosis, preoperative assessment, and prevention for complications of shunt surgery are made to promote a wide use of them. PMID- 18408357 TI - Cognition, continence and transfer status at the time of discharge from an acute hospital setting and their associations with an unfavourable discharge outcome after stroke. AB - BACKGROUND: Current demographic trends pose a major societal challenge due to the rising number of older people with chronic conditions such as stroke. The relative impact of various disabilities at the time of discharge from an acute unit on discharge outcome is poorly understood. OBJECTIVE: To examine the association between cognition, continence and transfer status at the time of discharge from the acute stroke unit and discharge destination. METHODS: A retrospective stroke register database study was conducted in an acute stroke unit in a UK hospital with a catchment population of 568,000. Consecutive acute stroke admissions between 1997 and 2003 who were discharged alive were identified and the likelihood of adverse discharge outcomes defined as institutionalization or a requirement for longer-term rehabilitation was estimated. RESULTS: A total of 2,521 discharges were analyzed (median length of hospital stay 8 days). The presence of confusion, urinary incontinence or the need for help with transfers at the time of discharge predicted a higher likelihood of an adverse outcome even after controlling for age, stroke subtype, premorbid Rankin score and length of hospital stay. The need for help with transfers appeared to be the most consistent and significant factor associated with an adverse outcome regardless of age, sex or stroke subtype across the sample distribution. CONCLUSION: The ability to transfer has a pivotal role in the clinical decision making of discharge destination after stroke. Understanding of the factors which may increase the potential for improving this ability after acute stroke could have an impact on clinical outcome. PMID- 18408358 TI - Skeletal muscle morphology and aerobic capacity in patients with obstructive sleep apnoea syndrome. AB - BACKGROUND: Despite the fact that patients with obstructive sleep apnoea syndrome (OSAS) often have symptoms at the level of skeletal muscle such as fatigue, the question of whether the structural, cellular and functional properties of limb skeletal muscles are affected has not been fully examined. OBJECTIVE: The aim was to examine physiological and muscular parameters in patients with OSAS and to assess the relationship between these parameters and the clinical symptoms. METHOD: Eighteen patients with OSAS and 16 controls participated. Aerobic capacity was assessed using a submaximal test. Fibre type distribution and fibre area were analyzed on muscle biopsies taken from the tibialis anterior. The microvascularization was assessed using the following parameters: (1) the number of capillaries per fibre (CAF), (2) CAF per fibre area (CAFA), (3) capillary to fibre perimeter exchange (CFPE) index, which represents the interface between muscle fibre and capillaries, and (4) length of capillary/perimeter of the fibre (LC/PF) index or capillary tortuosity, which represents the percent of muscle fibre perimeter in contact with the wall of the microvessel. RESULTS: The OSAS group had significantly lower predicted relative maximal oxygen uptake (p = 0.0047) which was inversely correlated to the apnoea/hypopnoea index (AHI; r = 0.6, p = 0.017). There was a significantly higher CFPE index for slow type I fibres (p = 0.007) and fast type II fibres (p = 0.0126) and a significantly higher LC/PF index for type I fibres (p = 0.0003) and type II fibres (p = 0.0285) in OSAS patients compared to controls. CONCLUSION: OSAS patients have a higher muscle microvascularization and a lower aerobic capacity than controls. Furthermore the aerobic capacity was inversely correlated to AHI. PMID- 18408359 TI - Validation of two instruments to assess technical bronchoscopic skill using virtual reality simulation. AB - BACKGROUND: In an era of increased emphasis on patient safety and competency based education, demonstration of significant variability in the teaching of flexible bronchoscopy has led to initiatives for new standardized curricula and assessment tools. OBJECTIVES: To evaluate the efficacy of such curricula and to assess bronchoscopic skill, 2 measuring instruments have been developed: the Bronchoscopy Skills and Tasks Assessment Tool (BSTAT) and the Bronchoscopy Step by-Step Evaluation Tool (BSET). We studied the validity and reliability of these 2 instruments. METHODS: Two independent testers simultaneously scored 22 volunteer participants at 3 levels [novice (n = 7), fellow (n = 8) and attending (n = 7)] on a virtual reality bronchoscopy simulator using the 2 instruments; each participant was tested twice, in 2 separate sessions. Intertester and test retest reliability were analyzed with intraclass correlations (ICC); ANOVA was used to assess concurrent validity based on the subjects' expected skill level. RESULTS: The ICCs between the testers were 0.98 for both the BSTAT and BSET. Comparison of the scores between the sessions showed high test-retest reliability by ICC (0.86 and 0.85 for BSTAT and BSET respectively), with a small yet statistically significant learning effect. The novice group's scores were lower than the fellows' and attendings' (p < 0.001) for both the BSTAT and BSET; the fellows' scores were consistently lower than the attendings' on both tests, yet the differences were not statistically significant. CONCLUSION: This validation study of 2 objective tests of bronchoscopic skill demonstrated high reliability and concurrent validity. These instruments can now be used to evaluate the effectiveness of new competency-based bronchoscopy curricula. PMID- 18408360 TI - Does change in gait while counting backward predict the occurrence of a first fall in older adults? AB - BACKGROUND: Dual-task-based assessment tests failed to establish a dependable relationship between dual-task-related gait changes and the risk of falls in the elderly. OBJECTIVE: The aim of this study was to examine whether changes in gait while counting backward could be associated with the occurrence of a first fall among older adults. METHODS: Walking while counting backward was investigated prospectively in a cohort of 187 older adults living independently in senior housing facilities. During enrollment, walking time, number of steps, and frequency of lateral line stepping-over and stops were measured while walking only and while walking with backward counting aloud. Information on the incident falls during the follow-up year was collected monthly. RESULTS: Walking time and the number of steps increased significantly under the dual-task condition compared to the single-task condition among fallers and non-fallers (p < 0.001). Compared to non-fallers, fallers had significantly lower scores in the Mini Mental State Examination (p = 0.029) and higher scores in the 15-item Geriatric Depression Scale (p = 0.003) and Timed Up & Go Test (p = 0.006) and increased walking time under both walking conditions (p = 0.030 for single-task condition and p = 0.007 for dual-task condition). After adjusting for these variables, depressive symptoms (adjusted OR = 2.6 with p = 0.041 and adjusted OR = 2.5 with p = 0.045 when walking time while walking only and walking with backward counting is considered, respectively) and walking time while walking only (OR = 2.3 with p = 0.032) were significantly associated with falls. CONCLUSION: Dual-task-related gait changes were poorly associated with the occurrence of a first fall and provided no additional predictive value compared to gait performance under a single task, suggesting that changes in basic clinical gait parameters while counting backward are unsuccessful to predict the first fall among older adults. PMID- 18408361 TI - Role of VGF-derived peptides in the control of food intake, body weight and reproduction. AB - VGF is a 68-kDa polypeptide synthesized in neuronal and neuroendocrine cells. It is cleaved into a number of smaller peptides which are stored in dense core vesicles and are likely to be secreted products. The VGF gene is expressed abundantly in the brain, and in peripheral endocrine tissues including the pituitary gland, the adrenal glands and the pancreas but also in the gastrointestinal tract in both the myenteric plexus and in endocrine cells. Several lines of evidence including observation of changes in hypothalamic VGF expression in catabolic states, a hypermetabolic phenotype in transgenic mice lacking VGF signalling, and demonstration of bioactivity of various peptide fragments have led to the view that VGF and/or its derived peptides are involved in the regulation of both energy balance and reproduction. PMID- 18408362 TI - Patient-reported outcomes of deferasirox (Exjade, ICL670) versus deferoxamine in sickle cell disease patients with transfusional hemosiderosis. Substudy of a randomized open-label phase II trial. AB - BACKGROUND/AIMS: There is increasing evidence demonstrating the value of transfusions in sickle cell disease (SCD). However, resultant iron overload can be life threatening if untreated. Chelation therapy with deferoxamine requires parenteral infusions that can negatively impact quality of life and adherence to treatment. METHODS: As part of a phase II trial, SCD patient-reported outcomes were evaluated. One hundred and ninety-five patients were randomized (2:1) to receive oral deferasirox (5-30 mg/kg/day) or deferoxamine (20-50 mg/kg, 5 days per week); 121 had previously received deferoxamine. RESULTS: At each time point, significantly more patients who had previously received deferoxamine were 'satisfied/very satisfied' with deferasirox, or found treatment to be 'convenient/very convenient' compared with deferoxamine (p < 0.001). In these patients, fewer hours were lost from daily activities with deferasirox than deferoxamine treatment. Most patients (77%) preferred deferasirox, and more were willing to continue taking deferasirox than deferoxamine at end-of-study (84 vs. 11%, respectively). CONCLUSIONS: Patients with SCD are therefore more satisfied with deferasirox, which has a lower impact on daily activities than deferoxamine. Given the high levels of satisfaction, it is likely that quality of life will be improved. These results also suggest that treatment adherence with deferasirox may be better than with deferoxamine, which should lead to improved long-term outcomes. PMID- 18408363 TI - Role of angiotensin II on dihydrofolate reductase, GTP-cyclohydrolase 1 and nitric oxide synthase expressions in renal ischemia-reperfusion. AB - BACKGROUND: The present study was conducted to investigate the role of renal ischemia-reperfusion (IR) and angiotensin II (ANG II) on mRNA and protein levels of renal dihydrofolate reductase (DHFR), GTP-cyclohydrolase 1 (GTP- CH 1), and endothelial and inducible nitric oxide synthase (eNOS and iNOS, respectively). METHODS: Male Wistar rats were sham operated or received IR (30 min occlusion, and reperfusion for 1 day). Each group was treated separately with water, angiotensin-converting enzyme inhibitor (ACEI) and ANG II receptor type 1 blocker (ARB) for 1 day before the sham operation or IR, and continuously for 1 day after the operation. The mRNA and protein levels were detected by RT-PCR and Western blot, respectively. RESULTS: IR decreased DHFR mRNA and protein levels (p < 0.01), both of which were restored by ACEI or ARB, whereas GTP-CH 1 expression was unaltered. IR suppressed eNOS dimer while enhancing the monomer (p < 0.01). IR augmented iNOS mRNA, total iNOS protein and iNOS monomer (all p < 0.01) which were attenuated by ACEI or ARB. CONCLUSION: Our study is the first to demonstrate that the heightened ANG II in IR, via stimulation of ANG II receptor type 1, suppresses DHFR and eNOS dimer, while activating both iNOS mRNA and protein levels. PMID- 18408364 TI - Lack of an association between Alzheimer's disease and the cystatin C (CST3) gene G73A polymorphism in Mainland Chinese. AB - Alzheimer's disease (AD) is a complex multifactorial disorder involving a number of genetic and environmental factors. Cystatin C (CST3), which belongs to the type II cystatin gene family, is a potent inhibitor of lysosomal proteinases. Immunohistochemical studies have demonstrated the colocalization of the b-amyloid (A-beta) and cystatin C peptides within arteriolar walls in the AD brain. The G73A polymorphism of the CST3 genemay be associated with AD development. To investigate a possible association between the CST3 G73A polymorphism and late onset AD (LOAD) in Mainland Chinese, we examined 281 LOAD patients and 376 healthy controls. All subjects were genotyped for CST3 and apolipoprotein E (APOE). There were no significant differences in the CST3 genotype or allele frequencies between the cases and the controls. Likewise, with the stratification of the APOE epsilon4 status, no statistical difference was observed between the cases and the controls. Our findings suggest that this polymorphism may not represent an additional genetic risk factor for LOAD in Mainland Chinese. PMID- 18408365 TI - Visual perceptual organization deficits in Alzheimer's dementia. AB - BACKGROUND: Deficits in visual processing are a prominent feature of Alzheimer's disease (AD), yet the cognitive and neuropathological mechanisms underlying these deficits are poorly understood. In the current study, we examined the hypothesis that perceptual organization is specifically impaired in AD compared to other dementias. METHODS: We examined perceptual organization with a psychophysically well-controlled measure of contour integration in patients with AD (n = 17), vascular dementia (n = 5), frontotemporal dementia (n = 5) and mild cognitive impairment (n = 10) and 11 age-matched healthy controls. RESULTS: Patients with AD differed significantly in their ability to detect contours comprised of Gabor elements as detection relied increasingly on long-range spatial interactions. Impairments in contour integration were particularly pronounced in AD patients with atrophy and gliosis of white matter in the occipital lobe. Deficits in perceptual organization were not found in patients with other dementias and participants with mild cognitive impairment. CONCLUSIONS: These results suggest that a subgroup of AD patients is characterized by a specific deficit in visual perceptual organization, which might reflect the impaired functional integrity of occipital cortico-cortical pathways. PMID- 18408366 TI - Estrogen receptor-alpha variants increase risk of Alzheimer's disease in women with Down syndrome. AB - BACKGROUND: Genetic variants that affect estrogen activity may influence the risk of Alzheimer's disease (AD). Two tightly linked polymorphisms (PvuII and XbaI) in the first intron of estrogen receptor 1 (ESR1), the gene for ER-alpha, have been reported to influence estrogen receptor expression and may influence the risk of AD. METHODS: We examined the relation of polymorphisms in ESR1 to the risk of AD in women with Down syndrome. The subjects (181 women with DS, 41-78 years of age) were followed at 14- to 18-month intervals. Information from cognitive assessments, caregiver interviews, medical record reviews and neurological examinations was used to classify dementia. Genomic DNA was genotyped for 5 single-nucleotide polymorphisms in the upstream region and the first exon/intron of the ESR1 gene. Their association with dementia risk was evaluated, adjusting for covariates. RESULTS: Women with at least 1 copy of the C allele at rs2234693 (PvuII) and those homozygous for the C allele at rs2077647 had an almost 3-fold increase in the risk of AD, compared with women without the C allele. The increased risks were independent of the apolipoprotein E genotype. CONCLUSION: These findings support a role for estrogen receptor activity in the development of AD in women with Down syndrome. PMID- 18408368 TI - The development of spinal cord anatomy. AB - A panel illustrating spinal cord injury in The Dying Lioness in the British Museum dates to 650 BC. This paper outlines the subsequent progression of knowledge of the anatomy of the spinal cord. The animal dissections of Galen are considered because his deductions persisted through the Dark Ages until the late 18th century. Anatomy advanced gradually to yield discoveries of the complex tracts and grey matter elements of the cord and their functions. Amongst many distinguished exponents, the works of Blasius, Huber, Vicq d'Azyr and Stilling are emphasised. PMID- 18408369 TI - Correlation of coronary artery atherosclerosis with atherosclerosis of the intracranial cerebral artery and the extracranial carotid artery. AB - BACKGROUND: Investigating atherosclerosis of the coronary artery in ischemic stroke patients is clinically important because comorbidity is relatively common in such patients. We studied the relationship of atherosclerosis of the coronary artery to atherosclerosis of the intracranial cerebral artery and extracranial carotid artery. Further investigation was performed for determining the factors independently associated with coronary artery atherosclerosis in ischemic stroke patients. METHODS: We consecutively recruited ischemic stroke patients who had no history of coronary artery disease, and they underwent vascular examination. Patient-based vascular assessment was performed with magnetic resonance angiography of the cerebral arteries and computed tomography coronary angiography. The factors independently associated with coronary artery stenosis (> or =50%) were obtained from the conventional vascular risk factors and cerebral arterial stenosis using the logistic regression model. RESULTS: Coronary artery stenosis was observed in 25.4% of the patients and this was associated with age (OR: 1.16, 95% CI: 1.03-1.30) and the presence of stenosis of the extracranial carotid artery (OR: 11.37, 95% CI: 1.88-68.75) after logistic regression analysis. Intracranial arterial stenosis was not independently related to coronary stenosis. CONCLUSION: Careful concern about coronary artery disease is needed when treating ischemic stroke patients who have atherosclerosis of the extracranial carotid artery. PMID- 18408370 TI - Ineffectiveness of folic acid supplementation against phenytoin-induced decrease in salivary immunoglobulin A concentration of epileptic patients. AB - AIMS: This study was designed to investigate if folate treatment is able to reverse the phenytoin-induced deficiency of salivary immunoglobulin A (IgA). METHODS AND MATERIAL: Twenty-five epileptic patients who had been under phenytoin therapy for at least the last 6 months were randomly selected and subjected to folic acid supplementation, 1 mg/day. The salivary IgA concentration of these patients was measured before and after 2 months of folic acid administration and compared with those of 10 healthy individuals. Independent and paired Student's t tests were used to analyze the effects of phenytoin and folic acid, respectively. RESULTS: Salivary IgA levels of patients receiving phenytoin (11.7 +/- 4.8 IU/l) were significantly (p = 0.039) lower than those of healthy controls (14.8 +/- 3.2 IU/l), but did not statistically (p = 0.541) differ from levels (11.8 +/- 4.6 IU/l) measured after 2 months of folic acid supplementation. CONCLUSIONS: According to these results, folic acid supplementation does not seem to have the efficacy to ameliorate phenytoin-induced salivary IgA hyposecretion. PMID- 18408371 TI - Sudden deafness related to posterior circulation infarction in the territory of the nonanterior inferior cerebellar artery: frequency, origin, and vascular topographical pattern. AB - AIMS: To investigate the frequency and origin of sudden deafness (SD) associated with the nonanterior inferior cerebellar artery (non-AICA) territory posterior circulation infarct(s), and the vascular topographical patterns of ischemic lesion associated with SD. METHODS: A total of 685 consecutive cases of vertebrobasilar ischemic stroke diagnosed by brain MRI from acute stroke registry were studied. RESULTS: Seven consecutive patients (1.0%) were identified as having SD as a symptom of non-AICA territory vertebrobasilar ischemic stroke. Audiological testing confirmed sensorineural hearing loss, predominantly cochlear in 4 patients and combined on the affected side cochlear and retrocochlear in 3 patients. Five patients had an isolated cerebellar infarct in the territory of the medial branch of the posterior inferior cerebellar artery and other 2 had infarct in the brainstem. In all, SD was on the side of infarct, as shown on brain MRI. Six patients had accompanied canal paresis to caloric stimulation on the side of SD. CONCLUSIONS: SD due to non-AICA territory infarction was mostly associated with infarct in the territory of the posterior inferior cerebellar artery. From the known variation of the anatomy of the internal auditory artery and results of audiovestibular evaluations, SD due to non-AICA territory infarct probably results from the damage to the inner ear. PMID- 18408372 TI - Alpha-1 antichymotrypsin gene signal peptide a/t polymorphism and primary intracerebral hemorrhage. AB - BACKGROUND/AIMS: Alpha-1 antichymotrypsin (ACT), a serine proteinase inhibitor, has been implicated in vascular pathology. The TT genotype of the ACT signal peptide A/T polymorphism has been reported to confer susceptibility to primary intracerebral hemorrhage (PICH). We conducted a prospective study to test possible association of ACT signal peptide A/T polymorphism with PICH in a Greek cohort with enough power (80%) to detect a twofold increase in the odds ratio. METHODS: We prospectively recruited 147 patients with PICH. ACT signal peptide A/T genotypes were determined in patients and 206 healthy, age- and sex-matched control subjects from the neurology outpatient clinic using the polymerase chain reaction restriction fragment length polymorphism method. RESULTS: Our study did not show an association between ACT signal peptide A/T polymorphism and PICH. We also failed to find any influence on age at onset, the location and volume of PICH as well as on clinical severity at admission or 6-month outcome. CONCLUSION: Our data failed to confirm an association between ACT signal peptide A/T polymorphism and PICH. However, we cannot exclude the possibility that the TT genotype confers susceptibility at less than a twofold increase. PMID- 18408373 TI - Clinical features of benign paroxysmal positional vertigo in Western Turkey. AB - BACKGROUND: It was the aim of this study to analyze the clinical manifestations, the incidence of each variant and the comorbid conditions of benign paroxysmal positional vertigo (BPPV) as well as the response to treatment. METHODS: One hundred and fifty-seven patients with BPPV were reviewed prospectively. An extensive neurotologic examination was performed. All patients were treated with an appropriate canalith repositioning maneuver (CRM). RESULTS: In 138 patients, the posterior canal (PC) was involved, in 14 patients, the horizontal canal (HC), in 2 patients, the anterior canal (AC), and in 3 patients, both the PC and HC. A history of head trauma was identified in 17 patients. In 1 patient sensorineural hearing loss on the affected side and in another bilateral peripheral vestibular loss was present. A history of migraine was reported in 21 cases. A resolution attributable to the first CRM was achieved in 132 patients. CONCLUSIONS: PC involvement was the most frequent type constituting 87.9% of all BPPV cases. HC, AC and mixed canal types were relatively rare constituting 8.9, 1.3 and 1.9% of the cases, respectively. Response to the first CRM was recorded in 84.1%. Association with migraine was recorded in 13.4% of the patients. PMID- 18408375 TI - Spontaneous dissections of the carotid arteries in a patient with recent coronary artery dissection. PMID- 18408374 TI - Clinical description of seizures in patients with systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the frequency of and risk factors for epileptic seizures in patients with systemic lupus erythematosus (SLE) in a large cohort series. METHODS: One thousand two hundred patients with SLE were analyzed. The type and frequency of risk factors for seizures associated with SLE were studied and compared with two other series reported in the literature. RESULTS: One hundred and forty-two patients had seizures. Seventy-five patients were studied with a mean follow-up of 5 years from the first seizure episode. Fifty-eight (77%) patients had tonic-clonic seizures, 9 (12%) complex partial seizures (PS), 5 (7%) simple partial motor seizures and 3 (4%) secondary tonic-clonic seizures. In 41 (54%) patients, the seizures occurred within the first year of the SLE diagnosis. Recurrence occurred in 40 (53%) patients, and was associated with PS in 14 (35%; p = 0.006) and time of seizures with SLE onset in 5 (12.5%; p = 0.05). Less than one third of the patients had positive antiphospholipid antibodies. A concurrent infection was present in 16 (21%) patients. CONCLUSIONS: Epileptic seizures were more common during the first year after SLE diagnosis. Neither infection nor antiphospholipid syndrome was associated with the occurrence of seizures. PMID- 18408376 TI - Multifocal necrotizing leukoencephalopathy mimicking sporadic Creutzfeldt-Jakob disease. PMID- 18408377 TI - Bilateral thalamic metastases in endometrial adenocarcinoma. PMID- 18408378 TI - Reversible vertebral artery stenosis following corticotherapy in giant cell arteritis. PMID- 18408379 TI - Some observations on the septum pellucidum. AB - The thin, vertically placed partition consisting of two laminae separated by a narrow chink constitutes the cavity of the septum pellucidum, known from the time of Sylvius. Traumatic lesions in boxers, rare congenital expanding cysts causing hydrocephalus, and a number of septo-optic dysplasias give clinical significance to the septum and its related cavum. PMID- 18408380 TI - Splenium. PMID- 18408381 TI - Straw Peter Syndrome - a literary mistake? PMID- 18408383 TI - A comparison of methods to detect recombination hotspots. AB - OBJECTIVE: A number of linkage disequilibrium (LD)-based methods have been developed to describe recombination and infer hotspots. We determine the correspondence between LDMAP and LDhat, and between LDMAP and LDhot by comparison with linkage maps and hotspots that have been verified by sperm typing. METHODS: Regression and variance analyses were used to compare LDMAP and LDhat with linkage maps. The location and intensity of hotspots inferred by LDMAP and LDhot were compared with fifteen verified hotspots. RESULTS: Despite different methodologies and assumptions, LDMAP, LDhat, and linkage maps are highly concordant. Closer inspection shows that LDMAP corresponds more closely with linkage maps across the genome and on sixteen chromosomes compared with LDhat. LDhot identified fourteen and ten of the verified hotspots using high and low density maps. In comparison, LDMAP identified all fifteen hotspots at high and low density. However, some significant discrepancies between sperm and LD-based recombination rates remain. CONCLUSIONS: Combining information from linkage and LDMAP to construct sex-specific high resolution linkage maps suggests that some of these discrepancies may be due to female recombination while others may relate to the age of hotspots. LDMAP based estimates between approximately 68,000 and approximately 112,000 hotspots in the genome with mean widths less than 4 kb. PMID- 18408385 TI - Cancer risk assessment and the genetic counseling process: using hereditary breast and ovarian cancer as an example. AB - While only a small proportion of cancers can be attributed to a hereditary susceptibility, identifying high-risk individuals plays an essential role in medical management and has a significant impact on the patient as well as their immediate and extended family members. This paper aims at increasing the medical professionals' knowledge of the components of a genetic counseling session, with particular attention toward identifying at-risk individuals and understanding the complexities of the testing process. In addition, tools are provided to assist in identifying these individuals in clinical practice and streamlining the referral process to a cancer genetics center. PMID- 18408386 TI - Experimental colitis is associated with ultrastructural changes in inflamed and uninflamed regions of the gastrointestinal tract. AB - OBJECTIVES: The objective of this study was to examine the ultrastructural changes in cell organelles such as mitochondria, endoplasmic reticulum (ER) and Golgi apparatus in inflamed colon and uninflamed ileum in colitic rats. MATERIALS AND METHODS: Colitis was induced in rats by intracolonic administration of trinitrobenzenesulfonic acid (TNBS). The animals were sacrificed on day 5 after TNBS administration and colonic and ileal samples were used for estimation of myeloperoxidase (MPO) activity, malondialdehyde (MDA) concentration, histologic examination and transmission electron microscopy. RESULTS: TNBS caused a significant reduction in body weight and an increase in MPO activity in colonic, but not in the ileal samples in animals with colitis. MDA levels were increased both in inflamed colon and the uninflamed ileal segments in colitis. Electron microscopy revealed swelling of mitochondria with broken cristae and disruption of the inner membrane. Colitis also caused fragmentation of the ER with loss of ribosomes and swelling of the Golgi apparatus with distended vesicles in both smooth muscle and epithelial cells in the ileal and colonic segments. These changes were absent in the control rats without colitis. CONCLUSIONS: These findings demonstrate ultrastructural deformities in both the mucosa and smooth muscle in inflamed and uninflamed regions of the gastrointestinal tract in experimental colitis. The structural changes in mitochondria are responsible for reduced ATP, while abnormalities in the ER and the Golgi apparatus may explain a generalized effect on protein synthesis, trafficking and targeting mechanisms, and may account for physiological changes seen in experimental colitis. PMID- 18408387 TI - Enhanced efficacy of amodiaquine and chlorpheniramine combination over amodiaquine alone in the treatment of acute uncomplicated Plasmodium falciparum malaria in children. AB - OBJECTIVE: To evaluate the comparative efficacy of amodiaquine (AMQ) alone and the combination of AMQ and chlorpheniramine (CP) in the treatment of acute uncomplicated malaria in children. SUBJECTS: Of the 110 children enrolled in the study, 103 with acute uncomplicated malaria, aged 6 months to 12 years, were evaluated using the 14-day modification of the WHO field test. The patients were randomized to 2 groups. Group 1 received supervised treatment with AMQ alone (10 mg AMQ base/kg daily for 3 days), while group 2 received supervised treatment with AMQ (same dose as group 1) plus CP (AMQCP) for 7 days. RESULTS: Both treatment regimens were well tolerated and no patient was withdrawn as a result of recurrent vomiting or drug-related adverse events. There was no significant difference in mean fever and parasite clearance times. The cure rates at day 7 were 90.2 versus 100% (rho = 0.027) for AMQ versus AMQCP, while the day 14 cure rates were 85.9 versus 98.1% for AMQ versus AMQCP, respectively (rho = 0.016). CONCLUSION: The combination of AMQ plus CP proved significantly more effective than AMQ alone in the treatment of acute uncomplicated falciparum malaria, most probably due to the enhancement of the antimalarial effect of AMQ by CP. The combination of AMQCP could be a better alternative to AMQ alone as a companion drug in artemisinin-based combination therapies. PMID- 18408388 TI - C-reactive protein levels in patients with chronic obstructive pulmonary disease: role of infection. AB - OBJECTIVE: To investigate the value of C-reactive protein (CRP) as a marker of chronic obstructive pulmonary disease (COPD) exacerbations or specifically bacterial exacerbations and to evaluate a correlation between raised CRP levels and other markers of inflammation in patients with an acute exacerbation (AECOPD). SUBJECTS AND METHODS: The medical records of patients with AECOPD were retrospectively analyzed. They were categorized according to the nature of sputum as mucoid or purulent and to the findings on chest radiographs as with pneumonia (PCOPD) or without pneumonia. Stable COPD (SCOPD) patients and a group of asymptomatic nonsmokers were also included in the study. RESULTS: All COPD patients (SCOPD: 30; AECOPD: 51; PCOPD: 32) and control subjects (30) were male. The mean CRP levels and WBC counts of the groups were PCOPD: 108.1 +/- 61.8 mg/l and 13.7 +/- 6.8 x 10(9)/l; AECOPD: 36.8 +/- 43.9 mg/l and 11.4 +/- 4.8 x 10(9)/l; SCOPD: 3.9 +/- 1.4 mg/l and 7.9 +/- 1.9 x 10(9)/l; control: 2.1 +/- 0.9 mg/l and 7.7 +/- 1.1 x 10(9)/l. The mean CRP level of AECOPD was statistically different from those of PCOPD and SCOPD (p = 0.0001, p = 0.002, respectively). The sensitivity and specificity of CRP to determine an acute exacerbation were 72.5 and 100%, respectively. Among the patients with AECOPD, 25 had purulent sputum and a mean CRP level of 46.4 +/- 48.6 mg/l, which is significantly higher than the CRP level (28.0 +/- 44.5 mg/l) of the 18 patients with mucoid expectoration (p = 0.015). Among the mucoid-expectorating subgroup, the patients with leukocytosis had significantly higher CRP levels than the patients without leukocytosis (p = 0.034). CONCLUSION: A high serum CRP value may indicate an infectious exacerbation in COPD patients and it correlates with sputum purulence and increased serum WBC counts. PMID- 18408389 TI - Tension-free vaginal tape and autologous rectus fascia pubovaginal sling for the treatment of urinary stress incontinence: a medium-term follow-up. AB - OBJECTIVES: To compare two different procedures, tension-free vaginal tape (TVT) and autologous rectus fascia sling, according to their medium-term subjective and objective outcomes and satisfaction rates in the treatment of urinary stress incontinence in women. SUBJECTS AND METHODS: One hundred women with type II urinary stress incontinence were randomized to be treated with either TVT or autologous rectus fascia sling. They were evaluated by means of a cough-induced stress test, 1-hour pad test, Incontinence Impact Questionnaire (IIQ), and urodynamic study. They were reevaluated postoperatively every 6 months, and the collected data of more than 1 year's follow-up were compared with preoperative assessments. RESULTS: All patients completed the full 6-month postoperative assessment. However, of the 100 patients, only 61 (25 in the TVT and 36 in the sling group) were followed for more than 1 year. Mean follow-up time was 38.5 and 40 months in the TVT and sling group, respectively. Objective cure was achieved in 22 (88%) of the TVT group and in 30 (83%) of the sling group (p = 0.78) using a cough-induced stress test, and in 76 and 75% of the women in the TVT versus sling group (p = 0.83), respectively, using a 1-hour pad test. Postoperative mean IIQ scores were 44.3 (range 35.5-61.5) and 48.5 (range 38.5-69.7) in the TVT versus sling group (p = 0.46). Five (20%) and 11 (30%) of the TVT and sling group, respectively, reported some changes in the voiding pattern or posture at more than 1 year's follow-up. CONCLUSION: There is no significant difference between the TVT and autologous rectus fascia sling procedures in the treatment of urinary stress incontinence at medium-term follow-up. There were changes in the voiding pattern for patients in both groups at more than 1 year postoperatively, which were not evident at early follow-up. PMID- 18408391 TI - Study motives and career choices of Iranian dental students. AB - OBJECTIVES: To investigate the study motives and career choices of Iranian senior dental students in relation to their background factors. SUBJECTS AND METHODS: During the spring term 2005, a questionnaire survey was administered to 327 senior dental students in seven randomly selected state dental schools in Iran. The questionnaire requested information on age, gender, parents' employment in dentistry, previous education and employment in dental hygiene, study motives, and career choices. Statistical analyses were made using independent sample t test, factor analysis, and binary logistic regression model. RESULTS: Based on the factor analysis, which explained 73% of the total variance, five motive dimensions were identified: altruism and intellectual challenges, characteristics of the profession, social status and security, other person's recommendation, and failure to be admitted to other study programmes. The mean for the 'characteristics of the profession' dimension was lower among the students with at least one parent employed in dentistry (p = 0.03). The 'altruism and intellectual challenges' dimension was reported to be more influential by the students with background in dental hygiene (32 students) compared to the others (p < 0.001). Engaging in postgraduate studies was the first career preference of 189 (70%) of the respondents. Those with a background in dental hygiene were less inclined to enter postgraduate courses (p < 0.001), but more eager to be employed in either the public or the private sector (p < 0.001), and to enter the community oral health and research field (p < 0.001) than the others. CONCLUSION: Personal characteristics and motives of the students play a major role in shaping their career preferences. PMID- 18408390 TI - Lateral cephalometric norms for adolescent Kuwaitis: soft tissue measurements. AB - OBJECTIVE: The purpose of this study was to establish lateral cephalometric soft tissue norms for adolescent Kuwaitis and to compare these norms to those of the common analysis systems. SUBJECTS AND METHODS: Digital lateral cephalograms were made of 82 Kuwaiti boys and 80 Kuwaiti girls, of mean age 13.27 +/- 0.42 and 13.21 +/- 0.43 years, respectively, with almost ideal occlusion. Anatomic landmarks were identified directly on the digital images. Linear and angular measurements were calculated electronically using the Dolphin (version 9) software package. RESULTS: The average subject in the sample had a smaller Z angle, a larger H angle, thicker soft tissue at point A and the pogonion, and an increase in most measurements for lip protrusion when compared to the norms of the common analysis systems. The ranges of the majority of facial and labial parameters were larger than those reported in the above-mentioned norms. Gender differences were limited to upper lip length. CONCLUSION: Our findings suggest that Kuwaiti norms for lip protrusion should be used when making the extraction decision in adolescent Kuwaiti orthodontic patients. The larger than previously documented variation of the majority of soft tissue parameters suggests a need for establishing different norms for different facial types. PMID- 18408392 TI - Magnesium sulphate therapy in women with pre-eclampsia and eclampsia in Kuwait. AB - OBJECTIVE: To evaluate the outcome of the use of MgSO4 therapy in women with severe pre-eclampsia in Kuwait from January 2002 to December 2004. SUBJECTS AND METHODS: The study involved 450 women managed at the Maternity Hospital in Kuwait with a blood pressure of 160/110 mm Hg and proteinuria of >0.3-5 g/24 h. A loading dose of 4 g MgSO4 was administered intravenously over 20 min and then the maintenance dose continued at 1 g/h for 24 h postpartum. Magnesium sulphate toxicity was monitored by urine output, deep tendon reflexes and serum magnesium levels and managed with an infusion of 10 ml of 10% calcium gluconate and cessation of magnesium infusion. Adjunct therapy included intravenous hydralazine 10 mg and labetalol 100 mg. The mode of delivery was determined after stabilizing the patient. RESULTS: The women included Kuwaitis (n = 200, 44.4%), Asians (n = 129, 28.7%) and other Arabs (n = 116, 25.8%) with a mean age of 29.7 +/- 6.7 years (primigravida: n = 233, 51.8%; other parities: n = 217, 48.2%). Antenatal complications included intra-uterine growth restriction (n = 136, 30.2%), oliguria (n = 39, 8.7%), haemolysis, elevated liver enzymes and low platelet count syndrome (n = 30, 6.6%), abruptio placentae (n = 20, 4.4%), eclampsia (n = 15, 3.3%), and preterm birth (n = 253, 55.2%). Caesarean section (n = 241, 53.6%) was the main mode of delivery. The perinatal mortality rate was 27 per 1,000. Magnesium sulphate toxicity observed as reduced tendon reflexes occurred in 14 (3.1%) patients and flushing, nausea and vomiting and blocked nostrils in 86 (19.1%). There was no association between adverse outcomes and maternal serum magnesium concentrations and no maternal mortality occurred. CONCLUSION: Magnesium sulphate was effective in preventing recurrence of eclamptic fits and safe for both mother and fetus. PMID- 18408393 TI - Serum level of intercellular adhesion molecule-1 in children with malignant lymphoma. AB - OBJECTIVE: To estimate the serum levels of soluble intercellular adhesion molecule-1 (s-ICAM-1) in children newly diagnosed with lymphoma and to correlate levels of s-ICAM-1 in lymphoma patients with clinical stage, pathological types, clinical and laboratory data and patient outcome. SUBJECTS AND METHODS: Thirty five children with newly-diagnosed malignant lymphoma (Non-Hodgkin's lymphoma, NHL: 23), Hodgkin's disease (HD: 12), and 8 apparently healthy subjects of matched age and sex taken as a control group were studied. For the patients and control group, the following tests were performed: complete blood count, and the following biochemical investigations: liver function tests, lactate dehydrogenase (LDH), and soluble ICAM-1 estimation using ELISA. In addition, for patients, pathological examination of lymph node biopsy for pathological grading, bone marrow aspiration and biopsy were done. Patients were observed for over 12 months or until death. RESULTS: Serum ICAM-1 increased more in HD and NHL than in the control group (p < 0.000); also s-ICAM-1 increased in advanced stages and high grade NHL (p < 0.008, 0.04, respectively). LDH levels were higher in patients compared to controls (p < 0.000). There was a positive correlation between high levels of s-ICAM-1 and increased levels of LDH in HD (r = 0.72, p < 0.008) and a positive correlation between high levels of s-ICAM-1 and increased ALT in NHL patients. A positive correlation between s-ICAM-1 levels and the presence of B symptoms in HD and NHL, and a positive correlation between elevated s-ICAM-1 levels and worse outcome in HD and NHL were detected. CONCLUSIONS: The data indicate that in children with malignant lymphoma, high serum levels of ICAM-1 correlated with tumor aggressiveness, and quantification of s-ICAM-1 levels may identify a subgroup of children with worse prognosis. Therefore, detection of s ICAM-1 levels in children with malignant lymphoma might represent an additional disease-associated marker for use in the clinical management of the patients. PMID- 18408394 TI - Cemented and cementless total hip replacement. Critical analysis and comparison of clinical and radiological results of 182 cases operated in Al Razi Hospital, Kuwait. AB - OBJECTIVE: In this study we present the results of a series of cemented Exeter and cementless Zweymuller implants. SUBJECT AND METHODS: Eighty-seven cemented and 95 cementless hip replacements for different hip pathologies were followed for an average period of 36 months for cementless and 60 months for cemented cases. Clinical results were calculated using the Merle d'Aubigne score. The orientation of the prosthetic components and the fixation of the cup and stem were analyzed. The clinical and radiological results were compared using statistical methods. RESULTS: In the average period of 36 months in cementless and 60 months in cemented hip replacements the clinical results improved significantly when compared with the preoperative score (p < 0.05). Sixty-seven cemented acetabular cups (77.1%) were in the desired position (30-50 degrees ) and 20 cemented cups (22.9%) were outside this range. Seventy-six cups (80%) were in the desired degree of abduction and 19 (20%) were outside this range. All cups except 1 were anteverted or neutral. Of the femoral stems, 173 were in the neutral position, 5 in the valgus and 4 in the varus position. Cemented cups were more commonly loose and cemented and cementless stems did equally well. No significant differences in rate of complications were found. CONCLUSION: Cementless acetabular implants had better clinical results and a lower loosening rate at 3 years of follow-up compared to cemented implants at 5 years of follow up. The cemented femoral implants were equally stable compared to the cementless ones. PMID- 18408395 TI - Whole blood assays to identify Th1 cell antigens and peptides encoded by Mycobacterium tuberculosis-specific RD1 genes. AB - OBJECTIVE: To identify Th1 cell-stimulating antigens/peptides encoded by the genes predicted in the Mycobacterium tuberculosis-specific genomic region of difference (RD)1, deleted in Mycobacterium bovis Bacille Calmette-Guerin(BCG), by using synthetic peptides and whole blood from tuberculosis (TB) patients. MATERIALS AND METHODS: Heparinized peripheral blood was obtained from culture proven pulmonary TB patients (n = 16) attending the Chest Disease Hospital, Kuwait. Whole blood was diluted with tissue culture medium RPMI-1640 and tested for Th1 cell stimulation using antigen-induced proliferation and interferon-gamma (IFN-gamma) secretion assays. The antigens included a peptide pool of 220 peptides covering the sequence of 12 open reading frames (ORFs) of RD1 (RD1(mix)), peptide pools of RD1 ORF5 (ORF5(mix)), ORF6 (ORF6(mix)) and ORF7 (ORF7(mix)), and individual peptides of ORF6 (P6.1-P6.6) and ORF7 (P7.1-P7.6). M. tuberculosis culture filtrate, cell walls and whole-cell M. bovis BCG were used as complex mycobacterial antigens. The results obtained with different antigens and peptides were statistically analyzed for significant differences using Z test. RESULTS: The complex mycobacterial antigens (culture filtrate, cell walls and M.bovis BCG) and RD1(mix) induced comparable (p > 0.05) positive antigen induced proliferation and IFN-gamma responses with whole blood from TB patients. However, the positive IFN-gamma responses induced by ORF6(mix) and ORF7(mix) were higher than ORF5(mix). Among the individual peptides, P6.4 and P7.1 of ORF6 and ORF7, respectively, induced the highest IFN-gamma responses, suggesting that these peptides represented the immunodominant Th1 cell epitopes of RD1 ORF6 and ORF7 in the patients tested. CONCLUSION: The whole blood assays with synthetic peptides are useful to identify Th1 cell antigens/peptides encoded by genes located in M. tuberculosis-specific genomic regions. PMID- 18408396 TI - Prostate small cell carcinoma and skin metastases: a rare entity. AB - OBJECTIVES: To report a rare case of small cell carcinoma of the prostate with unusual skin metastasis. CLINICAL PRESENTATION AND INTERVENTIONS: A 60-year-old was evaluated for difficulty in urinating. Abdominal computed tomography scans revealed a prostatic mass invading the surrounding tissues and multiple perirectal, periprostatic, para-aortic and pericaval lymph nodes. Needle biopsy specimens showed both small cell carcinoma and adenocarcinoma. He was treated with combination chemotherapy: cisplatin and etoposide and bilateral orchiectomy. After six cycles of the chemotherapy, disease progressed and the patient did not respond to salvage therapy; hence, palliative care was instituted. During the follow-up, papillary lesions were observed in the scrotal skin; biopsy showed metastatic small cell carcinoma. CONCLUSION: Small cell carcinoma of the prostate is an aggressive disease with a highly metastatic potential; but skin metastases are very uncommon. It has poor prognosis despite therapy. Management resembles that of small cell carcinoma of the lung. PMID- 18408397 TI - Headache as a first manifestation of Vogt-Koyanagi-Harada disease. AB - OBJECTIVES: To describe headache as an initial presentation of Vogt-Koyanagi Harada (VKH) disease. CLINICAL PRESENTATION AND INTERVENTION: A 56-year-old man initially presented with a 6-month duration of a continuous, diffuse headache of mild to moderate and sometimes of severe intensity. A CT of the brain was normal. Neurological examination was normal, including absence of symptoms and signs of meningismus. During the last 2 months, an intermittent eye pain, redness, and gradual loss of vision in both eyes was detected and subsequently he had tinnitus, malaise, nausea and mild meningismus. He was diagnosed as having VKH disease. Ophthalmologic examination revealed high intraocular pressure, requiring trabeculectomy with surgical iridectomy. The patient is now being treated with systemic steroids. CONCLUSION: VKH disease may initially present as sustained headache, without specific ophthalmologic symptoms and it should be considered in the differential diagnosis in patients with atypical but relentless headache. PMID- 18408398 TI - An unusual association between familial mediterranean fever and IgM nephropathy. AB - OBJECTIVE: To report a case with the diagnosis of IgM nephropathy and familial Mediterranean fever (FMF). CLINICAL PRESENTATION AND INTERVENTION: A 9-year-old boy was admitted to our hospital with recurrent abdominal pain since the age of 4 years. Laboratory investigations revealed a sedimentation rate of 88 mm/h, C reactive protein: 83.2 mg/l (0-10 mg/l), white blood cell count: 12,700/mm(3), fibrinogen: 622 mg/dl (200-400 mg/dl) and serum amyloid A: 186 mg/l (0-5.8 mg/l). Urinalysis revealed +2 proteinuria. A 24-hour urinary protein excretion was 12 mg/m(2)/h. M694V homozygous mutation was identified in exon 10. Percutaneous renal biopsy showed mesangial cell proliferation and increased mesangial matrix in the glomeruli, without amyloid accumulation. Immunofluorescence study showed IgM (+1) and C1q (+1) deposits. Treatment with 1 mg/day colchicine was started. Six weeks later, proteinuria had disappeared and the patient was asymptomatic. CONCLUSION: This case illustrates the unusual association of FMF with non-amyloid glomerulopathy. Glomerular diseases such as IgM nephropathy may be seen as a manifestation of FMF. PMID- 18408399 TI - Epilepsy in a young adult caused by L-2-hydroxyglutaric aciduria: a case report. AB - OBJECTIVE: To report the importance of a rare organic acid metabolic disorder, L 2-hydroxyglutaric aciduria, and its characteristic neuroimaging cerebral white matter abnormalities in a case of epilepsy. SUBJECT AND METHODS: A 19-year-old male presented with an 11-year history consisting of school failures, intellectual deterioration and generalized tonic-clonic convulsions. RESULTS: Neurological examination showed mental subnormality, mild dysarthria and bilateral pyramidal signs. Computed tomography and magnetic resonance imaging (MRI) of the brain showed characteristic white matter lesions, suggestive of L-2 hydroxyglutaric aciduria. The diagnosis of this disease was confirmed by elevated urinary concentrations of L-2-hydroxyglutaric acid. The epilepsy was partially controlled with antiepileptic drugs. CONCLUSION: This report indicates the importance of routine examination of urinary organic acids in children and young adults presenting with chronic encephalopathy and epilepsy with characteristic MRI white matter lesions. L-2-hydroxyglutaric aciduria should be considered as one of the differential diagnoses of epilepsy. PMID- 18408400 TI - Development of an effective breast and cervix cancer screening strategy in Mumbai: an impressive effort. PMID- 18408402 TI - Determinants of compliance in a cluster randomised controlled trial on screening of breast and cervix cancer in mumbai, India. 2. Compliance to referral and treatment. AB - OBJECTIVES: The study aims to investigate the efficacy of screening by low-cost technology in down-staging and reduction of mortality due to breast and cervix cancer. METHODS: The present trial is a community-based, cluster randomised controlled cohort study on screening for breast and cervix cancers (clinical breast examination and visual inspection of the cervix after application of 4% acetic acid). Univariate and multivariate logistic regression analyses are conducted to identify the predictors of compliance to referral among screen positive women and to treatment among cancer cases. RESULTS: The compliance to diagnostic investigations is 73% among screen-positive women referred for breast cancer and 79% among women referred for cervix cancer. Younger women, women working in service or being self-employed, school level-educated women, mother tongue Marathi, participation in screening in all 3 rounds and women referred as screen positive for cervix cancer had higher compliance to diagnostic investigations. The compliance to treatment completion is higher in women diagnosed with breast cancer, at 95%, compared to 86% for cervix cancers and 81% for cervix pre-cancers. CONCLUSIONS: Good compliance rates along with a proper system of referral, further investigations, confirmation of diagnosis and treatment as demonstrated in this trial are crucial for successful screening programmes. PMID- 18408401 TI - Determinants of compliance in a cluster randomised controlled trial on screening of breast and cervix cancer in Mumbai, India. 1. Compliance to screening. AB - OBJECTIVES: This study aims to investigate the efficacy of screening by low-cost technology in down-staging and reducing mortality due to breast and cervix cancer. METHODS: The present trial is a community-based, cluster randomised controlled cohort study on screening for breast and cervix cancers (clinical breast examination and visual inspection of the cervix after application of 4% acetic acid). Univariate and multivariate logistic regression analyses are conducted to identify the predictors of participation in screening. RESULTS: The average compliance is 71.43 and 64.93% for breast and cervix cancer screening, respectively, with the highest compliance in round 1. At the end of 3 screening rounds, 94 and 84% of the eligible women were screened at least once for breast and cervix cancer, respectively. Younger women, women from other than Hindu and Muslim communities, school level-educated women, women belonging to lower-income families, Marathi-speaking women, married women and women who had previously consulted for any breast or gynaecological complaints had higher compliance to participation in screening. CONCLUSIONS: Good compliance rates to screening have been demonstrated in the trial, reflecting acceptance of the study by the society, which has implications while translating the trial into a programme. PMID- 18408403 TI - F.E.S.S. (functional endoscopic sinus surgery) in China. PMID- 18408404 TI - Functional endoscopic sinus surgery in china. AB - The functional endoscopic sinus surgery (FESS) technique was introduced in China at the beginning of the 1980s and popularized gradually. Now its application has extended to the management of skull base lesions, orbital lesions and sinonasal tumors. This paper will review the application and relevant research of nasal endoscopic surgery and discuss its further perspectives in China. PMID- 18408405 TI - Endoscopic anatomic characterization of the frontal recess and its implications in frontal sinus surgery. AB - OBJECTIVE: This study was designed to describe the anatomic features of the frontal recess by transnasal endoscopy (nasoscope), to analyze its implications in endoscopic frontal sinus surgery and to discuss the issues that may be experienced during such operations. MATERIALS AND METHODS: The patients included in this analysis were adults with chronic sinusitis or nasal polyp (n = 301, 562 sides) hospitalized in our hospital from August 1998 to April 2001. Chronic frontal sinusitis was confirmed in 280 cases (479 sides) by coronal and axial CT scan. Patients with a previous surgical history were excluded from the analysis. The surgical outcomes of these patients and CT imaging data were retrospectively reviewed and analyzed. The drainage pattern of the frontal sinus was identified based on CT scans preoperatively. Endoscopic frontal sinus surgery was performed in 250 cases (421 sides). The anatomic features of the frontal recess under nasoscope were classified into 2 types. RESULTS: The coronal CT results confirmed the position of the frontal sinus ostium between the uncinate process and the middle turbinate in 203 sides (48.2%) of all operated patients and the ostium was found to be located between the uncinate process and the lamina papyracea in 218 sides (51.8%). According to the location of the frontal sinus ostium, we grouped the anatomic features of the frontal recess into 2 types. Type I was documented in 203 sides (48.2%) and type II was seen in 218 sides (51.8%). CONCLUSIONS: The anatomic features of the frontal recess under nasoscope and their classification are very important and helpful for endoscopic frontal sinsus surgery. The upper part of the uncinate process is a dependable anatomic landmark for the localization of the frontal sinus ostium in CT scan and endoscopic frontal sinus surgery. PMID- 18408406 TI - Effects of ephedrine on human nasal ciliary beat frequency. AB - The purpose of this study was to investigate the effects of ephedrine, an alpha,beta-adrenoceptor agonist, on the regulation of human nasal ciliary beat frequency (CBF). Changes in (cultured) human nasal CBF in response to 0.25, 0.5, 1.0 and 2.0% ephedrine were quantified by using high-speed (240 frames per second) digital microscopy combined with a beat-by-beat CBF analysis. At approximately 25 degrees C, we found that ephedrine induces an instant and moderate increase in human nasal CBF followed by an inhibitory response. With concentrations ranging from 0.25 to 2.0%, while the increment is independent of ephedrine concentration, the inhibitory effect is dependent on the concentration of ephedrine. Based on these results we propose that the clinically used concentration of ephedrine, 0.5%, has maximum stimulatory effect without obvious inhibitory effect on (cultured) human nasal CBF. PMID- 18408407 TI - The role of superantigens in chronic rhinosinusitis with nasal polyps. AB - Recent reports suggest that staphylococcal exotoxins, acting as superantigens, activate T cells with subsequent massive proliferation, thereby contributing to the etiology of chronic rhinosinusitis with nasal polyps (CRSwNP). The objectives of this study are (1) to demonstrate directly the presence of staphylococcal exotoxins in nasal mucosa and sinonasal polyp tissue, and (2) provide indirect evidence of the effect of superantigens on the T cells expressing the target of superantigen, i.e., the beta variable chain receptor (TCRBV) in polyp tissue and peripheral blood of patients with CRSwNP. Sinonasal polyp tissue and peripheral blood specimens were obtained from 37 patients with chronic rhinosinusitis (22 patients with bilateral nasal polyps, 15 without nasal polyps) and 12 normal subjects for comparative negative controls. Tissue specimens were assayed by enzyme-linked immunosorbent assay (ELISA) for the most common staphylococcal exotoxins (A-D) and toxic shock syndrome toxin type 1. Fresh tissue and blood samples were analyzed by flow cytometry to determine the expression of TCRBV. In the CRSwNP subjects 12 of 22 samples (54.54%) demonstrated reactivity for at least 1 staphylococcal exotoxin, while 2 of the 12 were positive for 2 toxins. There were no positive results in the CRS without nasal polyps or control groups. There was a clear trend of increased TCRBV expression in the ELISA-positive group both for tissue and blood specimens. Staphylococcal superantigens were present in the nasal cavity of patients with CRSwNP with a high percentage of TCRBV, which suggests the possibility of superantigens as etiological agents of CRSwNP. PMID- 18408408 TI - Multiplanar computed tomographic analysis of the frontal recess region in Chinese subjects without frontal sinus disease symptoms. AB - The purposes of this study were to determine the prevalence of frontal-ethmoidal cells and to evaluate variation in the superior attachment of the uncinate process in Chinese subjects. 202 normal Chinese subjects (404 sides) underwent spiral computed tomography and multiplanar reconstruction images were evaluated. Agger nasi cells showed a prevalence of 94.1%. Of all the frontal cells identified in 159 sides (39.6%) of frontal recesses, the prevalence of type I, type II and type III cells was 24.4, 7.0 and 8.2%, respectively. Suprabullar, frontal bullar and interfrontal septal cells were identified in 148 sides (36.6%), 36 sides (9.0%) and 25 subjects (12.4%), respectively. 244 uncinate processes (60.4%) had 1 superior attachment and the remainder (39.6%) had 2 superior attachments. The prevalence of terminal recesses was 89.1%. Our results characterized normal frontal recess pneumatization patterns in Chinese subjects. That, together with variation in the superior attachment of the uncinate process, emphasized the role of agger nasi cells and the uncinate process in endoscopic frontal sinus surgery. PMID- 18408409 TI - Clinical prognostic factors of chronic rhinosinusitis after endoscopic sinus surgery. AB - OBJECTIVE: We aimed to determine the clinical prognostic factors of chronic rhinosinusitis after endoscopic sinus surgery (ESS) in order to better guide the management of patients. METHODS: A total of 254 patients with chronic rhinosinusitis after ESS with a follow-up of 24 months were evaluated retrospectively. Clinical grouping, hyperplasia of the ethmoid bone, anatomic variations, allergy, polyps, previous sinonasal surgery and recurrent polyps were analyzed. RESULTS: The success rates were 64.4 and 91.9% with and without hyperplasia of the ethmoid bone, 87.6 and 86.4% with and without anatomic variations, 67.9 and 92.0% with and without allergy, 81.0 and 94.0% with and without polyps, 72.8 and 93.6% with and without history of surgery and 52.3 and 94.3% with and without recurrent polyps. In logistic multiple regression analysis, allergy (p = 0.0002; estimate, -4.2094), nasal polyps (p = 0.0007; estimate, -3.6393) and history of surgery (p < 0.0001; estimate, -5.0938) were found to reach a statistically significant level. CONCLUSIONS: We propose that allergy, polyps and history of surgery might be significant indicators of poor prognosis after ESS. PMID- 18408410 TI - Stages of nasal mucosal transitional course after functional endoscopic sinus surgery and their clinical indications. AB - OBJECTIVE: We aimed to explore the transitional process of sinus cavity mucosa after functional endoscopic sinus surgery (FESS) and divide it into different stages. METHOD: The morphological characteristics of sinus cavity mucosa after FESS were continuously observed. RESULTS: Over 90% of the nasal and sinus cavities got clean within 1-2 weeks, over 80% of the operative cavities experienced a reaction to mucosa removal or regenerative diseases within 3-10 weeks, such as mucosal edema, vesicles, granulation tissue, mini-polyps, fibrous hyperplasia and adhesion, which competed against the mucosa epithelialization process. After careful treatment, 90% of the nasal and sinus cavities finished epithelialization, and 60% of them completed epithelialization within 11-14 weeks after operation. CONCLUSION: The recovery process of mucosa after FESS can be divided into 3 stages: the stage of clean operative cavity, the stage of mucosal transition and the stage of complete epithelialization. Proper management at the 3 stages is very essential to the whole curable effect of FESS. PMID- 18408411 TI - Endoscopic endonasal surgery for clival chordoma and chondrosarcoma. AB - BACKGROUND: Conventional microsurgical approaches are limited to exposing the lateral and bilateral regions because of surgical trauma. Total removal of clival chordoma and chondrosarcoma is very difficult. This study was undertaken to determine the potential role of an endoscopic endonasal approach in improved management of clival chordoma and chondrosarcoma. METHODS: Seven patients with chordoma and 2 patients with chondrosarcoma treated by endoscopic endonasal surgery between November 2002 and February 2006 were studied prospectively. The mean age was 35 years. The follow-up periods ranged from 3 to 39 months. RESULTS: Total removal was achieved in 6 cases of chordoma and 1 case of chondrosarcoma, with subtotal removal in 1 case of chordoma and of chondrosarcoma. At the last follow-up, 7 patients had no evidence of disease and 1 was alive with disease. The chordomas had recurred in 1 patient after 5 months with subtotal removal. CONCLUSION: The use of nasoendoscopy to perform clival chordoma and chondrosarcoma surgery is not limited to merely the minimally invasive aspects. It provides better visualization of the deeper anatomical structures in the skull base and affords a means to 'look around corners'. This approach promises a simple and rapid access to the clivus. It is both a safe and efficient procedure. PMID- 18408412 TI - Endoscopic optic nerve decompression for the treatment of traumatic optic nerve neuropathy. AB - OBJECTIVE: We aimed to investigate the efficacy of endoscopic optic nerve decompression in patients with traumatic optic neuropathy. METHODS: We performed a retrospective analysis of 46 patients with traumatic optic neuropathy in the Shanghai Eye, Ear, Nose and Throat Hospital between March 2002 and September 2005. All patients were first treated with methylprednisolone for 6 days. Forty four patients (46 eyes) that did not improve with methylprednisolone treatment were offered endoscopic optic nerve decompression. RESULTS: In 38 eyes with no light perception vision preoperatively, 21 eyes (45.6%) had improvement in visual acuity. These patients had postoperative light perception in 17 eyes, hand movement in 3 eyes and 60/200 in 1 eye. Four of 5 eyes with light perception preoperatively had postoperative vision for hand movement in 2 eyes, finger counting in 1 eye and 20/200 in 1 eye. For 3 eyes with preoperative visual acuity of hand movement, the postoperative visual acuities were 60/200, 60/200 and 120/200. Neither worsening of vision nor major complications was encountered in our series. CONCLUSIONS: We conclude that endoscopic optic nerve decompression in experienced surgeons' hands can improve visual acuity in traumatic optic nerve neuropathy with minimal morbidity. Our results also demonstrate that even patients initially without light perception may benefit from optic nerve decompression. PMID- 18408413 TI - Endoscopic sinus surgery for the management of orbital diseases. AB - The indications for endoscopic sinus surgery (ESS) have been widely expanding since its introduction into sinus surgery. ESS has become an ideal method to manage certain orbital diseases and has the advantages of excellent visualization with minimal cosmetic and functional morbidity. In the Department of Otolaryngology of Taichung Veterans General Hospital from 1988 to 2005, 3,136 patients received ESS. Among them, a total of 108 patients received ESS for orbital diseases. These orbital diseases included orbital complications secondary to bacterial rhinosinusitis, fungal rhinosinusitis, skull base osteomyelitis and mucoceles, subperiosteal hematoma, Graves' disease, traumatic optic neuropathy and orbital blowout fracture. Our results showed that ESS is effective in the management of some of these orbital diseases. PMID- 18408414 TI - Dihydroartemisinin potentiates the cytotoxic effect of temozolomide in rat C6 glioma cells. AB - Gliomas are the most common primary brain tumor in adults, but the efficacy of chemotherapy is limited. Artemisinin and its analogs, such as dihydroartemisinin (DHA), can kill cancer cells via generating free radicals. In the present study, we determined whether DHA at low concentrations potentiates the cytotoxic effect of temozolomide in rat glioma C6 cells. We found that the IC50 values of DHA and temozolomide for cell viability were 23.4 and 560 micromol/l, respectively. The cytotoxic effect of temozolomide was enhanced by 177% at a nontoxic DHA concentration (1 micromol/l), and by 321% at a low-toxic DHA concentration (5 micromol/l). DHA substantially increased temozolomide-induced apoptosis and necrosis. The generation of intracellular reactive oxygen species (ROS) was increased by temozolomide combined with DHA at noneffective concentrations of both agents. Edaravone (20 micromol/l), a ROS scavenger, reversed the effects of temozolomide/DHA on both ROS generation and cell viability reduction. These results indicate that DHA at low concentrations potentiates the cytotoxic effects of temozolomide in C6 cells partly via generating ROS, suggesting a beneficial combination for the chemotherapy of gliomas. PMID- 18408415 TI - Characterization of binding affinity of CJ-023,423 for human prostanoid EP4 receptor. AB - In order to characterize the receptor binding pharmacology of CJ-023,423, a potent and selective EP4 antagonist, we performed a radioligand receptor binding assay under various assay conditions. An acidic (pH 6) and hypotonic buffer is a conventional, well-known buffer for prostaglandin E2 receptor binding assays. CJ 023,423 showed moderate binding affinity for human EP4 receptor under conventional buffer conditions. However, its binding affinity was greatly increased under neutral (pH 7.4) and isotonic buffer conditions. In this report, the binding mechanism between CJ-023,423 and human EP4 receptor is discussed based on the binding affinities determined under various assay conditions. PMID- 18408416 TI - Relationship between Theory of Mind and executive function in schizophrenia: a systematic review. AB - BACKGROUND: There is good agreement that Theory of Mind (ToM) and executive function are impaired to some degree in many patients with schizophrenia. However, few studies have set out to investigate as a primary aim the relationship between these 2 cognitive domains in the disorder. METHODS: A systematic review was carried out to identify all published studies in which tests of ToM and executive function were administered to individuals with schizophrenia, and in which the relation between scores was reported. RESULTS: The review revealed 17 relevant studies, of which 8 reported the relation between ToM and executive function using multivariate statistics. These all concurred in showing that ToM ability continued to predict an individual having schizophrenia (rather than being a control participant) once executive function was controlled for. CONCLUSIONS: There is very good agreement that ToM and executive function impairments in schizophrenia are independent of one another. Implications are discussed for the putative brain systems involved in the disorder and for its cognitive rehabilitation. PMID- 18408417 TI - A bibliometric study of the use of the classification and diagnostic systems in psychiatry over the last 25 years. AB - BACKGROUND: Our purpose was to examine the use of classification and diagnostic systems in the field of psychiatry (CDSP) from a bibliometric perspective, over the period 1980-2005. METHODS: We selected (in EMBASE and MEDLINE databases) documents that contained, in any of their sections, the descriptors 'psychiatr*', 'DSM*', 'ICD*', or 'diagnostic criteria',as well as other more specific descriptors. As a bibliometric indicator of production we applied Price's law. We also calculated the national participation index (PI) and correlated it with overall PI in biomedical and health sciences, and with PI in the discipline of psychiatry. RESULTS: We obtained 20,564 original documents; 15,743 referred to the Diagnostic and Statistical Manual of Mental Disorders (DSM) and 3,106 to the International Classification of Diseases (ICD). Our results indicate non fulfilment of Price's law, since scientific production on CDSP does not undergo exponential growth (correlation coefficient r = 0.9651, vs. r = 0.9927 after linear adjustment). Of the 10 journals with the highest impact factor in the field of psychiatry, the Journal of Clinical Psychiatry has the highest PI in the DSM subgroup (PI = 14.77), and the British Journal of Psychiatry in the ICD subgroup (PI = 1.54). The principal producer country is the United States (PI = 37.9), though in proportion to its production in the psychiatric field the ranking is headed by Finland. Only 10 countries, of the 20 major producers in health sciences, surpass their own PI in the field of psychiatry (Brazil, Italy, Japan, Austria, Spain, Germany, France, India, Switzerland, and China). CONCLUSIONS: Over recent years, the use of CDSP (basically the DSM or ICD) in the scientific literature has increased. Nevertheless, the abstracts to these studies, included in the principal databases, should always specify the diagnostic criteria employed, with a view to increasing information levels and reliability for the reader. PMID- 18408418 TI - Self-injurious behavior in adolescent girls. Association with psychopathology and neuropsychological functions. AB - BACKGROUND: Self-injurious behavior (SIB) is increasingly popular in psychically ill adolescents, especially in girls with posttraumatic stress (PTSD) and personality disorders. Adolescents with SIB frequently exhibit neurofunctional and psychopathological deficits. We speculated that specific neuropsychological deficits and temperamental factors could predispose patients to SIB and prospectively explored adolescent psychiatric patients with and without SIB in order to find out differences in psychopathology, and neuropsychological or temperamental factors. SAMPLING AND METHODS: Ninety-nine psychically ill adolescent girls with SIB, aged 12-19 years and treated at our clinic, were prospectively recruited during a period of 5.5 years (1999-2005). The clinical (ICD-10) diagnoses were mainly substance abuse, eating disorders, depression, PTSD and personality disorders. The control group was also prospectively recruited during the same period and consisted of 77 girls with similar diagnoses and ages but no SIB. All patients were subjected to the same selection of clinical and neuropsychological tests, mainly self-rating questionnaires and tests evaluating executive functions. RESULTS: Adolescent girls with psychiatric disease and SIB were more severely traumatized and depressed. They reported severe emotional and behavioral problems and deficits of self-regulation. In addition, their parents more frequently had psychiatric problems. Temperament, intelligence, investigated executive functions and presence of dissociative symptoms were not different in patients with and without SIB. CONCLUSIONS: We could not verify our primary hypothesis that SIB is related to specific neuropsychological deficits or temperamental factors. SIB was associated with traumatic experience, depression, problems of self-regulation and parental psychiatric disease. The prevention of SIB should therefore focus on improving affect regulation, the management of emotional distress and problem-solving strategies. PMID- 18408419 TI - Potential vulnerability markers within the affective domain in subjects at genetic and clinical high risk for schizophrenia. AB - BACKGROUND: Relative to ample high-risk studies on neurocognitive function, only a few high-risk studies have examined affective functioning components as possible vulnerability markers. In this study, we comprehensively assessed baseline affective functioning in subjects at clinical high risk (CHR) and genetic high risk (GHR) for schizophrenia, and healthy controls (HC), and compared the results to elucidate possible vulnerability markers in the affective domain. METHODS: We studied 3 groups of subjects: those with CHR (n = 28) or GHR (n = 28) and a HC group (n = 24). Affective-process- and affective-content related functioning were assessed using 5 emotion-related scales. RESULTS: In affective process, CHR subjects showed impairments in emotional awareness and mood repair, with some trend of impaired emotional expressivity as well as aggression control relative to either HC or GHR subjects, whereas GHR subjects showed only a trend of impairment in mood repair. In affective content, CHR subjects had less positive and more negative affect scores than the other 2 groups. CONCLUSIONS: These results correspond to previous findings of prodrome studies of schizophrenia and chronic schizophrenia and suggest that impaired mood repair and emotional awareness, as well as less positive and more negative affect may be potential candidates of vulnerability markers. PMID- 18408420 TI - Possession states in Northern Sri Lanka. AB - BACKGROUND: Possession states are still commonly seen in developing societies as acceptable cultural phenomena in normal persons as well as in those with psychiatric illness. 'Possession' is defined here as the experience of being taken over, controlled or occupied by another spirit or force. SAMPLING AND METHODS: This is a descriptive cross-sectional study of possession states among psychiatric patients, general population and popular adepts in Northern Sri Lanka, using semi-structured questionnaires and clinical observations. RESULTS: Thirty psychiatric patients were identified as having possession states. They were compared with 30 controls each from the general population admitted to a general hospital outpatient department and selected popular adepts in the community. The latter are individuals who are well known as having possession states. An analysis of social factors and other variables showed that education, marital status, age, employment, strength of belief, alterations in personality, past or family psychiatric history, previous exposure to similar phenomena, help seeking behavior and treatment outcome differed between the three groups. Religion of the subjects or recent changes in values showed no correlation with possession while monetary gain from possession states showed only a partial correlation. Western medical treatment was of value only when possession states were seen as part of psychotic illness. DISCUSSION: Possession is a spectrum of experiential and behavioral phenomena seen in culturally acceptable form in normal people, popular adepts, as well as manifestations of psychotic illness. Possession states which fit normal cultural stereotypes could, if necessary, be better managed by traditional methods. However, clinicians need to be familiar with culturally abnormal forms of possession which are manifestations of psychotic illness that benefit from western psychiatric treatment. PMID- 18408421 TI - [Familial aggregation of gastric cancer]. AB - Pursuit of the etiologies of familial aggregation of gastric cancer is a particular challenge in an endemic area like Japan. A typical case having truncation-type CDH1 germline mutation, which constitutes an entity of hereditary diffuse gastric cancer, has never been found in Japan or Korea. In addition to candidate gene approaches, painstaking efforts are now in progress using several genome-wide approaches including genome-wide association studies which require a few replications of several thousands of cases and controls. Gene-environmental interactions are also determined to find out the attributes for gastric cancer predisposition. PMID- 18408422 TI - [Certified medical oncologist]. AB - Certified medical oncologists are required to have excellent skills in chemotherapy, palliative care, second-opinion consultation, and the conducting of clinical trials. The Japanese Society of Medical Oncology has just started its certification system in 2006, and 126 clinical oncologists have been cumulatively certified. They have had an important role in facilitating better cancer practice, education and system innovation in their own institutions in cooperation with some government initiatives in Japan. PMID- 18408423 TI - [Board-certified radiation oncologists--what does a cancer treatment specialist mean ?]. AB - The Japanese Board of Cancer Therapy (JBCT) has started the board certification of general oncologists, who have a general knowledge for the treatment of cancer. Board-certified general oncologists could co-operate with board-certified specialists for specific cancer or organs so as to provide all cancer patients in the nation with high-quality treatment. The number of board-certified radiation oncologists in Japan is so small that they cannot meet the increasing demand for radiation therapy. An issue in the departments of radiation oncology of many medical schools is how to develop an attractive training program for radiation oncologists. We discuss the present status of board certified radiation oncologists. PMID- 18408424 TI - [Gann-chiryou Ninnteii--education of a fundamental (junior) oncology specialist]. AB - The development of new active agents, along with rapid progress in biomedical technology in recent decades, has resulted in major progress in cancer treatment, but at the same time it has made cancer management much more complex: Medical treatment must now be supplemented by various other types of care. Cancer patients and their families require the best care and support through the entire process, and optimal treatment for patients cannot be provided without an oncology health care team that includes various specialists such as medical-, radiation-, and surgical oncologists, and other professionals. The oncology education system in Japan, however, is unable to comply with this demand at present, although educational programs for highly trained and dedicated oncology experts are rapidly developing. Oncologists, whether subspecialized in medical, pediatric, radiation, or surgical oncology, must comprehend a formidable knowledge base and then effectively interact with medical colleagues in a wide range of disciplines in cancer care. Since the rapid improvement of this poor oncology environment is urgently required, the Japanese Board of Cancer Therapy has made a proposal to develop a primary certification system for oncology, Gannchiryou Ninnteii: This system provides education for candidates in basic principles common to management and treatment of malignant diseases, which can enhance the quality of oncology care and also maintain the standards for certification of various oncology specialists who are now developing in Japan. Candidates are required to pass the certifying examination after satisfactory completion of graduate education in a specialty and the following training programs as a fundamental oncologist. The first certifying examination is scheduled in January, 2008. The details for certification were reviewed. PMID- 18408425 TI - [Nursing certification system in cancer nursing]. AB - The Japanese Nursing Association (JNA) started an advanced nursing certification system and gave the first certification to certified nurse specialist (CNS) in cancer nursing in 1996. CNS is recognized for excellent nursing practice in a specific area, and for demonstrating six roles: excellent nursing practice, consultation, ethical coordination, education, coordination, and research activities. To receive CNS certification need nurse (or public health nurse or nurse midwife) qualification, master's degree, at least five-years clinical experience, and after that to pass a JNA certification examination. Certified nurse (CN) demonstrates three roles in high level of nursing practice by using matured nursing skills and knowledge, leadership, and consultation in a specific nursing area. CN need nurse qualification, at least fiveyears clinical experience, completing educational program at least 6 months, and passing a JNA certification examination. Both of CNS and CN requier certification renewal every five years. The number of people who get certification of CNS's in cancer nursing become 104, and of CN in cancer nursing area become 942 (chemotherapy 204, palliative care 420, cancer pain 267, breast cancer 51) in 2007. CNS's in cancer nursing activities are not understood broaden, because they have various work positions and activities, and are very few in Japan. But they are considered to be a change agent in cancer health care system. Also they will expand their activity setting. There is a prospect that CNS in cancer nursing will increase in number with nursing graduate school increasing. It shows that we will face some problems, for example educational contents or methods, certification system and so on. PMID- 18408426 TI - [Oncology pharmacy specialists in oncology]. AB - To provide effective and safe cancer treatment, the medical staff must form a team with patients and their family. Pharmacists have to be responsible for verifying chemotherapy prescription orders, mixing of anticancer drugs, management of adverse drug reactions, patient education, while providing drug information and participation in palliative care. To promote pharmacists who have acquired advanced knowledge and skills, The Japanese Society of Hospital Pharmacists started and educational program and established the certification of board-certified oncology pharmacy specialists (BCOPS) and board-certified pharmacists in oncology pharmacy (BCPOP). As of November 2007, 56 pharmacists were qualified as BCOPS. PMID- 18408427 TI - [Long- and short-term survivors after chemotherapy among patients with advanced or recurrent gastric cancer in our hospital]. AB - Since S-1 was approved in 1999, the response rate and the disease control rate of this chemotherapy for advanced or recurrent gastric cancer has been improved. Aa a result, we can plan for chemotherapy in patients with advanced or recurrent gastric cancer, because the median survival time (MST) with advanced or recurrent gastric cancer was over 20 months. We examined the characteristic findings of long- and short-term survivors in chemotherapy patients with advanced or recurrent gastric cancer. The prognosis for advanced gastric cancer is better than that for recurrent gastric cancer (p <0.02). In patients with only recurrent gastric cancer, the interval from surgery to recurrence is a significant prognostic factor. We thus show the results of chemotherapy for advanced or recurrent gastric cancer in our hospital and the characteristic findings in long- and short-term survivors for chemotherapy, and discuss effective planning of chemotherapy for each patient with advanced or recurrent gastric cancer. PMID- 18408428 TI - [Phase I/II study of combination chemotherapy with gemcitabine and UFT for advanced pancreatic cancer in a multi-center trial]. AB - The aim of this phase I/II study was to evaluate the tolerability and efficacy of combination chemotherapy with gemcitabine (GEM) and UFT for advanced pancreatic cancer. In phase I study UFT was given orally every day for 14 days and GEM was infused on day 1 and 8 at three dose levels (800, 900, 1,000 mg/m(2)/week) every 21 days. GEM 1,000 mg/m(2) and UFT 400 mg/m(2) did not reach the maximum tolerated dose. We decided that the recommended dose (RD) was GEM 1,000 mg/m(2)and UFT 400 mg/m(2). In phase II study 27 patients were enrolled and received GEM and UFT at RD. The tumor response rate was 17.6%, and the median survival was 221 days, which was very similar to that of GEM monotherapy. Due to adverse events, especially liver dysfunction, protocol therapy was discontinued in 12 patients. This study could not revealed the superiority of the GEM monotherapy. PMID- 18408429 TI - [Combined irinotecan (CPT-11) and nedaplatin (254-S) therapy for advanced and/or recurrent cervical cancer]. AB - We performed combination therapy with irinotecan (CPT-11) plus nedaplatin (254-S) for patients with cervical cancer. A total of 9 patients with cervical cancer (5 patients treated in neoadjuvant setting and 4 patients for recurrent disease) were administered 80 mg/m(2) of 254-Sintravenously on day 1 and 50 mg/m(2) of CPT 11 intravenously on day 1, 8 and 15. Treatment was repeated every 4 weeks. The average number of courses administered was 3.8 (range, 2-6). Grade 3 or 4 adverse events were leukopenia in 4 patients, thrombocytopenia in 1 patient and vomiting in 1 patient. The response rate was 40% in the neoadjuvant setting (2 PRs) and 75% in recurrent disease (3 PRs). PMID- 18408430 TI - [Cost-effectiveness analysis of 5-HT3 receptor antagonist drugs in cancer chemotherapy]. AB - Recently, ambulatory treatment centers (ATC) are markedly increasingboth in number and scale. It is therefore important to consolidate an efficient therapeutic system. A decrease in both treatment time and waitingtime leads to not only the improvement of the quality of life (QOL) for patients but also the efficient use of personnel and running costs for medical institutions by reducingthe bed occupation rate. In ATC, 5-HT3 receptor antagonists are extensively used for high emetic risk patients. However, their high cost and prolonged treatment causes one of the problems in improvingthe efficiency of the therapeutic system when they are administered by intravenous infusion. Amongthe 4 types of 5-HT3 receptor antagonists (injections) currently available in Japan, azasetron is the only drugthat is not designated as a powerful drug and that can be administered by bolus intravenous infusion. In this study, we investigated azasetron and granisetron from the standpoint of pharmacoeconomics with a simulation model using the results of clinical studies in Japan. Accordingto the results of cost-effectiveness analysis, therapeutic and time costs per patient for azasetron 10 mgand granisetron 2 mg (calculated in consideration of both medical institutions and patients) was 8,219 and 10,193 yen, respectively. This gap was attributable to the time loss due to the difference in administration methods. The result suggests that this time loss is more significant not only for patients but also for medical staff than the loss attributable to the drugcost. Furthermore, the bolus intravenous infusion of azasetron is considered superior to the non-bolus intravenous infusion of granisetron from a pharmacoeconomic standpoint. It is desirable to choose the appropriate administration method of 5 HT3 receptor antagonists in various chemotherapy regimens for the purpose of reducingthe treatment time and promotingthe efficiency of the therapeutic system at ATCs. PMID- 18408431 TI - [Usefulness of perospirone for the prevention of nausea related to opioid administration]. AB - PURPOSE: In our department, prochlorperazine, a D2 receptor antagonist, was previously administered to prevent nausea related to the introduction of opioids. However, we always had to consider extrapyramidal symptoms. Since August 2006, perospirone has been administered to all patients who have commenced oral opioid therapy, to prevent nausea. In this study, we investigated its usefulness in the prevention of nausea. METHODS: Perospirone (4 or 8 mg, once a day) was combined in all patients in whom opioid therapy was introduced with oxycodone at 10 mg/day. We retrospectively examined the presence or absence of nausea within 1 week. RESULTS: Oxycodone-related nausea occurred in 1 of 25 patients. There were no adverse reactions such as extrapyramidal symptoms. CONCLUSION: These results suggest the usefulness of perospirone for the prevention of nausea related to the intro- duction of opioids. PMID- 18408432 TI - [A case of advanced esophageal cancer with large liver metastasis successfully treated with FAP therapy]. AB - A 70-year-old man with dysphagia was diagnosed as advanced esophageal cancer by a primary doctor, and he was admitted to our hospital for treatment in February, 2004. The pretreatment diagnosis was basaloid squamous carcinoma, Mt area, T4 (aorta) , N2 (No. 107) , M1 (liver), Stage IVb performed systemic chemotherapy by FAP (5-fluorouracil ( 5-FU)+doxorubicin (DXR)+cisplatin (CDDP) ) from March, 2004. After 4 courses, the local tumor almost entirely disappeared, and the liver metastasis was obviously reduced. We continued chemotherapy afterwards. As of March 31, 2007, he had local lesion CR and metastatic lesion PR. It is very important to perform FAP repeatedly, for local and metastatic lesion of esophageal cancer while maintaining the patient's general condition and avoiding adverse events. PMID- 18408433 TI - [Complete response in a case of advanced esophageal cancer treated with docetaxel/5-FU/CDDPand S-1/docetaxel as neoadjuvant chemotherapy]. AB - A 64-year-old woman with advanced esophageal cancer underwent chemotherapy with docetaxel/5-FU/CDDP (DFP). Adverse reactions were severe nausea and general fatigue, so the patient decided to discontinue DFP therapy. The treatment was changed to S-1/docetaxel. Adverse reactions were not so severe, so she could receive 1 course of the medication completely. After the treatment, the primary lesion showed a partial response, so we performed surgery. In the resected specimen, no malignant cell could be seen microscopically. Though the advanced esophageal cancer was regarded as a systemic disease, the appropriate combination of chemotherapy and surgery proved effective. PMID- 18408434 TI - [A case of advanced gastric cancer treated with S-1/CDDP combination chemotherapy through jejunostomy]. AB - Recently, although chemotherapy for advanced gastric cancer has been proving more highly effective, no standard chemotherapy for gastric cancer has been established. We administered S-1 combined with cisplatin (div) to a patient with advanced gastric cancer who underwent a jejunostomy because of swallowing difficulties (PS 4) due to cerebral infarction. The overall response of this chemotherapy was a partial response (PR) for 14 months. We concluded that the administration of S-1 combined with cisplatin (div) through a jejunostomy can improve the nutrition management and the quality of life (QOL) of a patient with advanced gastric cancer who is incapable of oral intake. PMID- 18408435 TI - [A case of effective weekly paclitaxel administration for advanced gastric cancer associated with diabetes mellitus -about the control of blood glucose when dexamethasone for prophylaxis against paclitaxel-associated hypersensitivity reactions to the diabetic has been administered]. AB - A 69-year-old woman had been treated with insulin for diabetes over the last 8 years. Distal gastrectomy (D2) was undertaken for StageIV stomach cancer. CA19-9 showed marked increases after surgery, but returned to normal after administering S-1. After 12 cycles, the treatment was discontinued due to hepatic disorders, and the clinical course was monitored. Weekly paclitaxel therapy was initiated as second-line therapy when CA19-9 rose again to 467 U/mL. Marked efficacy was noted after completion of one cycle. A total of 23 cycles were conducted. CA19-9 returned to normal, and the patient remains recurrence-free. In the treatment with paclitaxel, pre-treatment with dexamethasone (20 mg each time) is made to prevent hypersensitivity reactions. Since the total dose becomes too large in weekly treatment, however, treatment was initiated at 12 mg in our patient, and the dose was reduced stepwise to 8 mg, 4 mg and 2 mg. At the same time, the dose of insulin at bedtime and before breakfast the following morning was increased in increments of 2 units. This made it possible to maintain good control of blood glucose levels and minimize changes in HbA1c. This experience suggests that the dosage regimen needs refinements in diabetic patients such as a reduction in the dose of steroids and increases in the dose of insulin in long-term treatment. PMID- 18408436 TI - [A case of unresectable advanced gallbladder cancer successfully treated by oral S-1 and hepatic arterial infusion (HAI) of low-dose CDDP therapy]. AB - The patient was a 64-year-old woman. Oral S-1 and hepatic arterial infusion (HAI) of low-dose CDDP therapy were started for unresectable advanced gallbladder cancer associated with liver metastasis and numerous lymph node metastases. Marked regression of the liver metastasis and lymph node metastases was observed by this treatment, and upon completion of the second course they had almost completely resolved. The tumor marker values also converted to negative. We report a case in which oral S-1 and HAI of low-dose CDDP therapy was effective against advanced gallbladder cancer associated with liver metastasis and multiple lymph node metastases. PMID- 18408437 TI - [A case of ruptured common hepatic arterial pseudoaneurysm and penetration into duodenum related with hepatic artery infusion chemotherapy]. AB - In a 64-year-old male patient, liver metastasis recurred after surgery for transverse colon cancer. He received 6 courses of continuous intra-arterial infusion therapy with a catheter for hepatic intra-arterial infusion, which was retained by the GDA-coil technique. In April 2005, the patient suddenly developed hematemesis, and he was hospitalized for emergency treatment. Upper gastrointestinal endoscopy on the day after hospitalization revealed no distinct source of bleeding. Since he had massive hematemesis again on hospital day 4, endoscopic examination was repeated, which revealed that the metallic coil for embolization retained in the gastroduodenal artery protruded from the superior wall of the duodenal bulb with bleeding. Emergency angiography revealed that the bleeding arose from rupture of a common hepatic artery aneurysm. Hemostasis was induced by embolization of the region ranging from the right and left hepatic arteries to the entire common hepatic artery aneurysm with the metallic coil. The present case revealed serious complications of the hepatic intra-arterial chemotherapy and intra-arterial retention of the catheter. PMID- 18408438 TI - [Effective treatment with oral administration of UFT and leucovorin (Uzel) in a patient with liver metastasis and peritoneal dissemination from cecal cancer]. AB - A 70-year-old woman had been diagnosed with advanced cecal cancer with liver metastasis. She had initially undergone a right hemicolectomy for cecal cancer. Although secondary surgery for liver metastasis was performed 2 months after initial surgery, the intra-operative findings showed peritoneal dissemination of diaphragm and ligament teres and liver metastasis, making curative resection impossible. Therefore , combination oral administration of UFT and LV (Uzel) was started (UFT 300 mg/day, LV 75 mg/day, 4 weeks of therapy followed by a 1-week treatment break). Two months after 4 courses, the liver metastasis had markedly diminished and CEA was within the normal range. A good complete response of the liver metastasis was achieved. In conclusion, this treatment was convenient and effective. PMID- 18408439 TI - [Multiple hepatic metastases and ovarian metastases of colon cancer responding to combined therapy with S-1 and CPT-11--a case report]. AB - We report a patient with multiple hepatic metastases and ovarian metastases of transverse colon cancer treated by combination of S-1 and CPT-11. The patient was a 51-year-old woman with cancer of the transverse colon and multiple hepatic metastases. She had undergone surgery. Resection of the transverse colon and left ovary was performed because left ovarian metastases were found during the operation. After the operation, the patient was given chemotherapy with S-1 (120 mg/body on days 1-14) and CPT-11 (150 mg/body on day 1). After completion of 11 courses of chemotherapy, abdominal CT scans revealed that the LDAs of the liver had disappeared, so the patient was judged to have achieved CR. No adverse event was observed. This case suggests that the combination of S-1 and CPT-11 may be an effective regimen for advanced colon cancer with multiple hepatic metastases. PMID- 18408440 TI - [Two cases of recurrent colorectal cancer treated successfully with folinate/tegafur/uracil (UFT/LV) chemotherapy on an outpatient basis]. AB - We report two cases of recurrent colorectal cancer in two patients who were treated successfully with a combined oral chemotherapeutic agent folinate/tegafur/uracil (UFT/LV) dosage. The first case was a 74-year-old woman who underwent Hartmann operation for colon cancer perforation. One year and 7 months after surgery, a local recurrence was found on the CT scan and endoscopy. It ended in exploratory laparotomy though we were operated on. Chemotherapy using 5-fluorouracil/Leucovorin (5-FU/LV) was performed six times. Sequentially, UFT/LV internal use was managed at our outpatient clinic. This patient has been living without side effects in the first three postoperative years, and without increase in tumors. The second case was a 65-year-old woman who underwent abdominoperineal resection of the rectum for rectal cancer. The histologic stage of disease aggravation of the patient was stageI. Post operatively, 2 years and 6 months later, we recognized a metastasis node on the lung upper right lobe of the patient and her CA19-9 value climbed dramatically. The patient took UFT/LV during outpatient visits to the hospital, the lung node shadow disappeared two months later, and CA19-9 decreased, too. The patient is living without a cancer recurrence now. UFT/LV treatment is one of the effective modalities against recurrence of colorectal cancer from outpatient treatment while maintaining QOL. PMID- 18408441 TI - [A case of lumbar metastases from rectal cancer successfully treated by UFT plus oral leucovorin (LV) therapy]. AB - A 63-year-old man had undergone a low anterior resection for rectal cancer with multiple liver metastases. Oral UFT (450 mg/day) administration alone was started after the operation. After 6 months post operatively, the patient was diagnosed as anastomosis recurrence because of ileus by abdominal X-ray. Transverse loop colostomy was performed by emergency surgery. After surgery, he suffered from paraplegia for lumbar vertebrae metastases. UFT+LV therapy was started. After chemotherapy a significant reduction of the lymph node and liver metastases and an apparent decrease in CEA and CA19-9 were observed. The patient left the hospital and showed no signs of tumor exacerbation for three months. The patient died of aggravation of primary disease afterwards. The therapy was safe and effective, and has successfully maintained the quality of life (QOL) of this patient. PMID- 18408442 TI - [A case of complete response in supraclavicular lymph node involvement from cancer of the sigmoid colon to FOLFOX4 therapy after laparoscopy-assisted colectomy]. AB - We reported a case of sigmoid colon cancer with left supraclavicular lymph node involvement that completely responded to FOLFOX4 therapy following laparoscopy assisted colectomy (LAC). A 69-year-old female presented with left supraclavicular tumor and was diagnosed as sigmoid colon cancer. Abnormally high CEA (128 ng/mL) and CA19-9 (59.3 U/mL) levels were detected in serum. She was operated palliatively by LAC, and FOLFOX4 therapy was initiated. The serum CEA and CA19-9 values regained their normal level after 5 and 4 courses of FOLFOX4, respectively. After 5 courses, metastatic nodes disappeared on her CT. After 12 courses no abnormal accumulations were detected on FDG-PET. Antitumor efficacy was judged as CR. Grade 1 neutropenia and grade 2 peripheral neuropathy were noted, but no other serious adverse reactions occurred during FOLFOX4 therapy. CR has been maintained at present. She has been treated with FOLFOX4 therapy without oxaliplatin to prevent neuropathy. Establishment of maintenance therapy following CR to FOLFOX4 is awaited. PMID- 18408443 TI - [Modified FOLFOX6 in a patient on hemodialysis with metastatic colorectal cancer]. AB - Since pharmacokinetics in patients undergoing hemodialysis differs from that in patients with normal renal function, chemotherapy for a hemodialysis patient should be considered with due care. We administered chemotherapy of modified FOLFOX6 to a patient on hemodialysis with inoperable metastatic colorectal cancer, and measured his plasma concentration of total platinum and non-protein bound platinum. Since there is no reported case of oxaliplatin use in patients on hemodialysis so far, we evaluated whether it could be safely used for such patients. We made a dose escalation study with 40, 50, 60, 70 and 85 mg of oxaliplatin, and evaluated the pharmacokinetics at each dose. AUC was 5.67-10.21 mg/L x h. The dialysis removal rate was 84.0%. Although this patient could accept it relatively safely without any severe side effect, the optimal dosage and the timing of hemodialysis for inoperable metastatic colorectal cancer patients should be determined by a further study using more cases. PMID- 18408444 TI - [A case of omental mesothelioma presenting with laminar thickening of omentum appearances of diffuse malignant peritoneal mesothelioma]. AB - A man in his 60's with no apparent history of asbestos exposure was admitted to our hospital with a chief complaint of abdominal fullness. CA125 levels in serum and ascites were veryhigh. Enhanced abdominal CT confirmed a large amount of ascites, inflexible intestinal canal, short mesentery and laminar thickening of the omentum. SPIO-enhanced MRI-T1WI slightly enhanced the thickened omentum. These T1WI images largely matched portal-phase contrast CT images. Furthermore, the thickened omentum was clearly visualized by lowering the signal for the liver and spleen by SPIO and by suppressing the ascites signal by fluid-attenuated inversion recovery (FLAIR). Gascintigraphy confirmed Ga accumulation in the same areas. Chemotherapy was ineffective, and the patient died of liver metastasis in February 2006. Autopsy confirmed biphasic malignant peritoneal mesothelioma. The involvement of asbestos is clear in the onset of malignant peritoneal mesothelioma. Therefore, it is possible that the patient unintentionally inhaled asbestos. Hence, when levels of CA125 in serum and/or ascites are high, it is important to differentiate malignant peritoneal mesothelioma from primaryserous papillary carcinoma of the peritoneum. Here, we experienced a case of biphasic diffused omental mesothelioma. While studies have documented laminar thickening of the omentum by abdominal incision, this is thought to be first case in Japan in which laminar thickening of the omentum was detected on diagnostic imaging. Laminar thickening of the omentum and short mesentery are thought to be characteristic features of diffused peritoneal mesothelioma when subjective symptoms appear. In addition, Gascintigraphy and FDG-PET are useful auxiliary diagnostic tools. In the future, we hope to differentiate epithelial, sarcomatous and biphasic types based on imaging findings. PMID- 18408445 TI - [A case of HER2-positive metastatic breast cancer responding to trastuzumab plus gemcitabine combination therapy]. AB - A 60-year-old woman was admitted to the hospital with left thigh pain. She had undergone mastectomy and axillary lymph node dissection for right breast cancer (T3N2M0) five years and two months earlier. The pathological diagnosis then was invasive ductal carcinoma with axillaryly mph node metastases. Hormone receptors and HER2 status were negative and positive (3+), respectively. The patient received adjuvant chemotherapy and radiotherapy, but bone metastases appeared 18 months after surgery. Although trastuzumab-combination chemotherapy with taxane and/or capecitabine was given, bone metastases in thoracic vertebra resulted in incomplete paralysis in both legs. She underwent thoraco-lumbar vertebral fixation 10 months before admission. A PET/CT revealed multiple bone metastases in the left femur as well as vertebrae, and CEA rose markedly. She received radiotherapy and trastuzumab monotherapy in addition to bisphosphonate. Temporarily, CEA decreased, but because recurrence nests were recognized in the supraclavicle and mediastinum after the eight-month treatment, trastuzumab monotherapy was followed by trastuzumab plus vinorelbine combined therapy. This regimen markedly reduced CEA after three months, but it rose again over the following three months. As S-1-combined therapy was not effective, trastuzumab+gemcitabine (1 g/week and two weeks on/one week off) combined therapy was started. CEA decreased markedly after 4 cycles, and FDG accumulation in the recurrence region was markedly improved. The adverse event during this treatment was minor, and PS was sufficiently maintained. These results suggest that trastuzumab plus gemcitabine combination therapy is effective for HER2-positive metastatic breast cancer. PMID- 18408446 TI - [A case of elderly breast cancer achieving partial response by letrozole with stable disease to anastrozole as neoadjuvant endocrine therapy]. AB - An 85-year-old postmenopausal woman had multiple lesions in her right breast. Core needle biopsy showed solidtubular carcinoma with positive hormone receptor (ER+++, PgR-). She refused surgery and was then treated with anastrozole as neoadjuvant endocrine therapy. After seven months of the treatment, all lesions were increased or unchanged. The clinical response was judged as stable disease. Letrozole was chosen for the next therapy. After ten months, reductions in the tumors were noted, and the clinical response was judged a partial response. No side effects were observed. These two drugs are nonsteroidal aromatase inhibitors, and it is considered there is a cross-resistance between them. However, several studies have revealed letrozole was significantly more potent than anastrozole in inhibiting aromatase activity in vitro and in inhibiting total body aromatisation in vivo. This may be related to a difference in effect between the two drugs. It is suggested that letrozole as second-line endocrine therapy may be effective for postmenopausal breast cancer even after anastrozole failure. PMID- 18408447 TI - [Prolonged survival in a patient with human herpesvirus-8-negative primary effusion lymphoma after combination chemotherapy with rituximab]. AB - Primary effusion lymphoma (PEL) is a unique clinicopathological entity usually associated with human herpesvirus-8 (HHV-8) infection. It occurs almost exclusively in human immunodeficiency virus (HIV) -infected individuals. We presented a rare case of HIV-negative PEL in an elderly HHV-8-negative patient who developed cardiac tamponade due to pericardial effusion. The patient was treated with rituximab and cyclophosphamide, doxorubicin, vincristine and prednisolone (CHOP). This disease generally has a poor prognosis; however, this patient achieved complete remission and remains without signs of disease 30 months after the last treatment. Because most HIV-negative and HHV-8- negative PEL cases show pan-B-cell markers, there is considerable usage of rituximab, though its optimal usage for PEL is unclear. To the best of our knowledge, there have been five reported cases where rituximab treatment has been used against HIV negative and HHV-8-negative PEL. The clinical courses of these cases were relatively good without specific adverse effects. HIV-negative and HHV-8-negative PEL appears to be a reasonably new clinicopathological entity. While further investigation will of course be needed, the use of rituximab is worth considering for treatment of such patients. PMID- 18408448 TI - [A case of liver metastasis of rectal cancer demonstrating complete response for three years to 5-FU/l-leucovorin]. AB - We report a case in which 5-fluorouracil/l-leucovorin (5-FU/l-LV) combination therapy was remarkably effective for non-resectable advanced rectal cancer with multiple liver metastasis. A 68-year-old man complaining of severe abdominal distension and abdominal pain was diagnosed as having ileus due to rectal cancer. We established a diagnosis of non-resectable rectal cancer with multiple liver metastasis and therefore performed only rectal colostomy. Systemic chemotherapy with 5-FU/l-LV was scheduled for a total of 22 courses postoperatively. After the chemotherapeutic regimen, a CT scan and colonofiberscopy revealed the primary lesions had disappeared, and a histological examination of biopsy confirmed that the patient had achieved complete response (CR). PMID- 18408453 TI - Liver disorders. PMID- 18408454 TI - Hepatobiliary pathology. AB - PURPOSE OF REVIEW: Studies are reviewed from the past year concerning the histopathology of liver and biliary diseases and their pathogenesis. RECENT FINDINGS: Several cases of acute hepatitis E showed portal and periportal hepatitis, with polarization of neutrophils to the interface region and lymphocytes more centrally in the portal tracts. Transfection of hepatitis C virus into cultured fetal hepatocytes resulted in sustained growth of 50-90 nm diameter viral particles. The ductular reaction in nonalcoholic steatohepatitis appears to evolve with fibrosis in response to hepatocyte replicative senescence. Hepatocellular release of hepcidin is critical for iron homeostasis in a manner analogous to pancreatic insulin for glucose homeostasis; this 'endocrine' focus was elaborated in an overview of hemochromatosis. Specific microscopic features of liver-cell adenomas combined with genetic analysis for hepatocyte nuclear factor 1alpha and beta-catenin mutations allows differentiation into four variants. Steroid-sensitive biliary strictures resembling primary sclerosing cholangitis but with increased serum immunoglobulin G4 and infiltrating immunoglobulin G4-positive plasma cells ('immunoglobulin associated cholangitis') are part of a spectrum of disorders including autoimmune pancreatitis and inflammatory pseudotumor. SUMMARY: Pathologic features of viral hepatitis C and E, immunohistochemistry for the ductular reaction and malignant liver tumors and several systemic disorders are among recent important pathology studies. PMID- 18408455 TI - Inherited metabolic disease of the liver. AB - PURPOSE OF REVIEW: Elucidation of metabolic pathways for copper and iron improved our understanding of Wilson disease and genetic hemochromatosis. Some inherited liver diseases are now characterized by protein-folding mutations, including Gaucher disease, cystic fibrosis and ZZ alpha1-antitrypsin deficiency. Studies now focus on associations between glycogen storage disease, hepatic adenoma formation and transformation to hepatocellular carcinoma. Continued progress in the study of the diagnosis, natural history and treatment of inherited liver diseases is the subject of this review. RECENT FINDINGS: Further understanding of metabolic pathways for iron and copper have led to a search for factors that modify phenotypic expression of Wilson disease and genetic hemochromatosis. Hepcidin plays a key role in modulating iron uptake in iron-overload disorders and new studies elucidate hepcidin regulation. For glycogen storage diseases, studies on the natural history and hepatocellular transformation necessitate tumor surveillance and possible early transplantation. A better understanding of genetic and nongenetic modifiers in ZZ alpha1-antitrypsin deficiency and other disorders of protein misfolding will improve our ability to manage these patients. SUMMARY: Recent discoveries in iron, copper and glycogen metabolism advance our ability to diagnose and treat inherited metabolic diseases of the liver. Some of these important findings are detailed in this review. PMID- 18408456 TI - Drug-induced liver injury in 2007. AB - PURPOSE OF REVIEW: To summarize the pertinent literature on the causes, epidemiology, prevalence, clinical features, evaluation and mechanisms of drug induced liver injury reported during 2007. RECENT FINDINGS: Although the frequency of drug-induced liver injury remains low, new data from the Centers for Disease Control and Prevention confirm that of the approximately 1600 new acute liver failure cases annually, acetaminophen hepatotoxicity accounts for 41%; among children with acute liver failure, acetaminophen was the second most common cause. Antimicrobials lead the list of non-acetaminophen causes of drug-induced liver injury. In Asia, herbal compounds are the most common causes of the condition. Pravastatin was shown to be safe in patients with nonalcoholic fatty liver disease or chronic hepatitis C. The US Food and Drug Administration issued a draft guidance document on the premarketing clinical evaluation and stopping rules of drug-induced liver injury signals, including Hy's Law cases in clinical trials. SUMMARY: The year 2007 brought with it several reminders of the importance of drug-induced liver injury in the clinical trial as well as the clinical practice setting. There is additional evidence that statin drugs may be used safely in patients with chronic liver disease. Comments received by the US Food and Drug Administration to finalize their guidance document are eagerly awaited. PMID- 18408457 TI - Autoimmune liver disease. AB - PURPOSE OF REVIEW: To review studies that improve the diagnosis and treatment of autoimmune hepatitis and extend understanding of its pathogenic mechanisms. RECENT FINDINGS: Black patients have more advanced disease and poorer outcomes than white patients. Genome-wide DNA microsatellite techniques have identified multiple regions that may confer susceptibility or resistance to the disease. Preferential inactivation of one parentally-derived X chromosome may favor autoreactivity in women. Acute and chronic hepatitis of undetermined cause can respond to corticosteroid therapy and represent autoantibody-negative autoimmune hepatitis. Outcomes can be improved by continuing therapies until resolution of all features and by early identification of problematic patients with the Model for End Stage Liver Disease. Serum levels of B-cell activating factor correlate with laboratory indices of liver injury. Tacrolimus and mycophenolate mofetil are promising therapies for problematic patients, and the antigenic targets of atypical antibodies to liver/kidney microsome may lead to diagnostic tests for de novo autoimmune hepatitis after liver transplantation. SUMMARY: Ethnic background and genetic predisposition affect the occurrence and outcome of autoimmune hepatitis. Susceptibility and resistance factors across the human genome underscore the genetic complexity of the disease. Outcomes can be improved by better use of current regimens and further evaluation of action-specific immunosuppressive agents. PMID- 18408458 TI - Update on viral hepatitis: 2007. AB - PURPOSE OF REVIEW: This is a concise review of recent developments in the field of viral hepatitis, based on publications between December 2006 and November 2007. RECENT FINDINGS: Hepatitis A vaccine has similar efficacy to immune globulin as postexposure prophylaxis. Entecavir is a potent antiviral agent with a low rate of drug resistance in nucleoside-naive chronic hepatitis B patients but it is not as effective in lamivudine-refractory patients. A combination of adefovir and lamivudine is preferred to adefovir monotherapy for lamivudine refractory hepatitis B patients. Two orally administered hepatitis C protease inhibitors, telaprevir and boceprevir, were shown to have antiviral activity in hepatitis C genotype 1 patients. A 16-week course of pegylated interferon and ribavirin resulted in a lower rate of sustained virologic response compared with the standard 24-week course. Patients with hepatitis C-related cirrhosis who achieved sustained virologic response to antiviral therapy remain at risk for hepatocellular carcinoma. A novel recombinant hepatitis E vaccine was shown to be safe and effective in preventing infection. SUMMARY: Advances have been made in the prevention of hepatitis A and hepatitis E. It is likely that specifically targeted antiviral therapies for hepatitis C will be available in the next few years. PMID- 18408459 TI - Hepatocellular carcinoma: latest developments. AB - PURPOSE OF REVIEW: This review primarily focuses on new developments in the field of hepatocellular carcinoma. RECENT FINDINGS: Potential preventive strategies in the development of hepatocellular carcinoma are being recognized. Novel molecular markers identified may aid in the diagnosis of early hepatocellular carcinoma in patients with chronic hepatitis C virus. Prognostic information gained by preoperative tumor biopsy is being investigated. Treatment of early hepatocellular carcinoma with resection versus primary or salvage transplantation continues to be debated. Expansion of selection criteria beyond the Milan criteria appears feasible. The role of living donor liver transplantation in hepatocellular carcinoma will require further study to determine the risk of recurrence. Improvements in chemoembolization with drug eluting beads appear promising. SUMMARY: Further insight into the pathogenesis of hepatocellular carcinoma will result in the continued evolution of our approach and management of the disease. Tailored therapies based on tumor biology are needed to improve treatment response and ultimately patient survival. PMID- 18408460 TI - Recent advances in nonalcholic fatty liver disease. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize recent advances in defining the clinical features, pathophysiology, natural history, and treatment of nonalcoholic fatty liver disease. RECENT FINDINGS: Nonalcoholic fatty liver disease is present in approximately 30% of the US population. A histologic grading and staging system has been developed and validated. Nonalcoholic fatty liver disease increases the risk of developing the metabolic syndrome. The presence and severity of nonalcoholic fatty liver disease correlates with the severity of obesity, fat distribution, age, and presence of other features of the metabolic syndrome. Fifteen to 20% of subjects with nonalcoholic steatohepatitis develop cirrhosis. Hepatic steatosis is associated with an increase in both free fatty acid delivery to the liver for re-esterification and increased de-novo lipogenesis. Several mechanisms of hepatocyte injury and death including free fatty acid toxicity, increased free cholesterol, cytokine-mediated injury and activation of the unfolded protein response have been defined. While many therapeutic targets have been identified and pilot studies performed, a definitive treatment for nonalcoholic steatohepatitis remains to be established. SUMMARY: Nonalcoholic fatty liver disease is a widely prevalent disease that is critically linked to insulin resistance and the metabolic syndrome. While much new information on the pathogenesis and natural history of nonalcoholic steatohepatitis is available, an effective therapy remains to be established. PMID- 18408461 TI - Alcohol and the liver. AB - PURPOSE OF REVIEW: To update the reader with advances in epidemiology, genetics, detection, pathogenesis and therapy of alcohol-related liver disease. RECENT FINDINGS: Ill-health due to alcohol abuse is improving in some nations but deteriorating in others. Oxidative and nitrosative stress are key to the pathogenesis of alcoholic liver disease, and there is now greater emphasis than previously on their development and role of cytochrome P450 2E1, on mitochondrial stress and disruption, (including elucidation of mitochondrial protection mechanisms) disturbance of signaling pathways and involvement of extrahepatic mediators like adiponectin. Treatment of alcoholic liver disease has stagnated, but transplantation is still favored and debated for end-stage cirrhosis. SUMMARY: Basic and clinical research into the mechanisms of alcoholic liver disease is making headway, but has yet to produce safe and effective therapies for alcoholic hepatitis and for reversing cirrhosis. PMID- 18408462 TI - Liver transplantation: an update 2008. AB - PURPOSE OF REVIEW: Expansion of the donor pool for liver transplantation is a priority. Management of hepatitis C and hepatocellular carcinoma has focused on decreasing recurrence rates after transplantation. RECENT FINDINGS: Expansion of the pool of donors has focused on live donor liver transplantation and extended criteria donor grafts. The results of live donor liver transplantation are equivalent to those of deceased donor liver transplantation. The use of extended criteria donor grafts has increased significantly. The results are associated with decreased graft survival with the use of grafts that have multiple factors considered as extended criteria for transplantation, particularly in high-risk individuals such as critically ill recipients. Judicious matching of extended criteria donors with recipients is essential to reduce waiting list mortality without reducing posttransplantation survival. The role of pretransplant ablation therapy for hepatocellular carcinoma is evolving to reduce tumor progression and dropout on the list as well as to influence posttransplant recurrence rates. Antiviral and immunosuppressive strategies in reducing the severity of hepatitis C virus recurrence are discussed as is retransplantation for the disease. SUMMARY: Expansion of the donor pool with the use of extended criteria donors and live donor liver transplantation is a major challenge. Transplantation for hepatocellular carcinoma and hepatitis C virus relapse are major areas of research. PMID- 18408463 TI - A strategic vision to promote advances in biliary tract disorders. PMID- 18408465 TI - Novel endoscopic approaches for assessing biliary tract diseases. AB - PURPOSE OF REVIEW: Biliary endoscopy offers both diagnostic and therapeutic value in complex clinical situations. This review addresses the latest advances over the past year in endoscopic approaches of biliary tract diseases. RECENT FINDINGS: Specifically, we focus on the latest findings on endoscopic retrograde cholangiopancreatography for the evaluation of biliary strictures. In addition, key studies have demonstrated the enhanced role of cholangioscopy, photodynamic therapy in cholangiocarcinoma, and biliary stent technology. SUMMARY: The following review focuses on the latest advancements in the field of biliary endoscopy. Pivotal studies were selected to highlight some of the current investigations in therapeutic endoscopic retrograde cholangiopancreatography as well as knowledge gaps for future research. PMID- 18408464 TI - Cholangiocarcinoma: lessons from Thailand. AB - PURPOSE OF REVIEW: To present the background of liver fluke-associated cholangiocarcinoma in Thailand focusing on recent epidemiological data and pathogenesis of this bile duct cancer. RECENT FINDINGS: More systematic tumor registration in Thailand nowadays uncovers new high-incidence areas that are confined to not only the northeastern part but also some provinces in northern Thailand. The link between the liver fluke, Opisthorchis viverrini, and cholangiocarcinoma, particularly in terms of cellular and molecular pathogenesis, is further elucidated. SUMMARY: Thailand is still the country with the highest incidence of cholangiocarcinoma in the world. Liver fluke induces chronic inflammation leading to oxidative DNA damage of the infected biliary epithelium and malignant transformation. Eradication of the fluke and identification of high risk populations are urgently needed. PMID- 18408466 TI - Genetics of biliary tract diseases: new insights into gallstone disease and biliary tract cancers. AB - PURPOSE OF REVIEW: Chronic biliary diseases are due to complex interactions between environmental and genetic factors. Here we summarize the current knowledge of genetic factors that contribute to common biliary diseases, focusing on gallstones and carcinogenesis, and review the recent association studies. RECENT FINDINGS: Since most studies were based on small sample sizes, replication of the findings is mandatory. Recently a large twin study confirmed a genetic predisposition to gallstones and a genome-wide association scan identified the hepatocanalicular cholesterol transporter ABCG8 as the common susceptibility factor for gallstone disease. Genetic studies in patients with cholangiocarcinoma indicate that genes controlling the metabolism and transport of xenobiotics or modulating chronic inflammation may determine individual susceptibility. SUMMARY: Genetic studies have identified the first susceptibility factors for gallstones and biliary tract cancers, but most results have yet to be replicated. In the future, genome-wide studies in different populations are likely to identify the entire set of genes contributing to chronic biliary diseases. Since the disease phenotypes result from the manifestation of susceptibility factors under the influence of environmental triggers, the discovery of these genes will open avenues to control environmental challenges and lead to novel strategies for risk assessment ('gene signatures') and prevention. PMID- 18408467 TI - Targeted therapies for cancer of the gallbladder. AB - PURPOSE OF REVIEW: Adenocarcinomas of the gallbladder are uncommon, aggressive tumors with poor survival. This review summarizes advances in understanding the biology of gallbladder cancer. RECENT FINDINGS: Published response rates of adenocarcinomas of the gallbladder to chemotherapy are less than 30% and no survival benefit has been demonstrated from palliative systemic therapy. New information on the molecular carcinogenic mechanisms of these malignancies, combined with findings from animal models, may lead to improved treatment for patients. SUMMARY: Improved understanding of the molecular carcinogenesis of adenocarcinomas of the gallbladder, coupled with the availability of novel molecularly 'targeted' chemotherapeutic agents, may improve outcome for patients. PMID- 18408468 TI - An update on primary sclerosing cholangitis. AB - PURPOSE OF REVIEW: Primary sclerosing cholangitis is a chronic cholestatic liver disease characterized by strictures of the biliary tree complicated by cirrhosis and cholangiocarcinoma. It is immune mediated, although the precise aetiology remains unknown. RECENT FINDINGS: The research into aetiology, genetic associations, pathogenesis, epidemiology, diagnosis of cholangiocarcinoma and medical treatments are discussed. SUMMARY: Multiple gene polymorphisms and human leucocyte antigen haplotype associations with primary sclerosing cholangitis have been investigated. Common inflammatory bowel disease associated polymorphisms and ulcerative colitis associated human leucocyte antigen haplotypes are not associated with primary sclerosing cholangitis. Biliary epithelial cells may mediate their own destruction by exaggerating innate and adaptive immune responses to bacterial products in the liver. The natural history of large and small duct primary sclerosing cholangitis has been reviewed. Positron emission tomography may be a useful adjunct to current imaging modalities in the pretransplant assessment of patients to exclude cholangiocarcinoma. Ursodeoxycholic acid remains the most studied medical treatment for primary sclerosing cholangitis; pilot studies suggest a possible role for tacrolimus and silymarin, however further studies are required. PMID- 18408469 TI - Successful liver transplantation for hilar cholangiocarcinoma. AB - PURPOSE OF REVIEW: The purpose of this review is to present recent data demonstrating the success of neoadjuvant therapy followed by liver transplantation for hilar cholangiocarcinoma. RECENT FINDINGS: The most recent outcomes of a protocol involving neoadjuvant therapy followed by liver transplant for hilar cholangiocarcinoma, placed in context with the most recent data regarding outcomes for surgical resection, are discussed. Data involving the observation and management of an increased rate of vascular complications following this aggressive protocol are described. Additionally, factors which may predict disease recurrence along with the implications of these data on organ allocation policy are reviewed. SUMMARY: For patients with early stage hilar cholangiocarcinoma which is unresectable or arising in the setting of primary sclerosing cholangitis, neoadjuvant chemoradiotherapy followed by liver transplantation provides excellent survival benefit. Outcomes are notably superior to other treatment options, including resection. The protocol, however, requires extensive involvement and expertise from multiple specialties to ensure success. PMID- 18408470 TI - Immunoglobulin G4-associated cholangitis. AB - PURPOSE OF REVIEW: Immunoglobulin G4-associated cholangitis is a recently identified clinical entity characterized by infiltration of immunoglobulin G4 bearing plasma cells in bile ducts and other affected tissues. This review is focused mainly on the most recent studies published during the last 2 years that have increased our understanding of this clinical entity. RECENT FINDINGS: Investigations concerning the immune mechanisms, liver histology in immunoglobulin G4-associated cholangitis and reports of clinical and radiological features of the condition as well as medical therapy are discussed. SUMMARY: Immunoglobulin G4-associated cholangitis was found to be characterized by overproduction of T helper 2 cells, and regulatory cytokines (interleukin-10 and transforming growth factor-beta) were upregulated in the affected tissues. Immunoglobulin G4 immunostaining in a range of clinically involved tissues in patients with immunoglobulin G4-associated cholangitis and autoimmune pancreatitis was found to be important for diagnostic purposes, which is probably most useful in cases with normal immunoglobulin G4 levels. It has become apparent that multiple measurements of immunoglobulin G4 levels in patients with clinically suspected immunoglobulin G4-associated cholangitis are important, revealing abnormally high levels in patients with normal immunoglobulin G4 on initial testing. Most patients respond to steroids initially but disease relapse seems to be common. A significant proportion of primary sclerosing cholangitis patients have elevated immunoglobulin G4 levels. PMID- 18408471 TI - Gallbladder polyps in primary sclerosing cholangitis: not so benign. AB - PURPOSE OF REVIEW: The increased risk of malignancy of the biliary tract and colon observed in primary sclerosing cholangitis raises concerns as to whether standard guidelines for the practical handling of polypoid lesions of the gallbladder apply to this group of patients. The present review aims to summarize existing knowledge on this topic. RECENT FINDINGS: Two recent studies reported adenocarcinoma in approximately 60% of the polypoid lesions of the gallbladder in patients with primary sclerosing cholangitis. One of the adenocarcinomas in the first of these studies, as well as two reported adenocarcinomas in a recent case series, occurred in lesions less than 1 cm. Cholecystectomy is normally not advocated for lesions less than 1 cm, but may be justified in patients with primary sclerosing cholangitis. In several studies, foci of dysplasia were also detected in separate adenomas and regions of the gallbladder not primarily affected by cancer. Concurrent dysplasia and malignancies of the bile ducts and colon have also been reported, and suggest that an increased awareness of malignancy is warranted in patients once gallbladder neoplasia has been detected. SUMMARY: The increased risk of malignancy in primary sclerosing cholangitis also applies to polypoid lesions of the gallbladder. Cholecystectomy and intensified screening for dysplasia of the bile ducts and colon may be advisable in primary sclerosing cholangitis patients with neoplasia of the gallbladder. PMID- 18408472 TI - Bibliography. Current world literature. Liver. PMID- 18408474 TI - Cryoglobulinemia and renal disease. AB - PURPOSE OF REVIEW: Cryoglobulinemia occurs in a variety of clinical settings including lymphoproliferative disorders, infection and autoimmune disease. The worldwide pandemic of hepatitis C virus infection has resulted in a significant increase in its extrahepatic complications including cryoglobulinemia and renal disease. Here we review the types of cryoglobulins, mechanisms of cryoglobulin formation, links between hepatitis C virus and renal disease, and current approaches to therapy. RECENT FINDINGS: The prevalence of cryoglobulinemia in hepatitis C virus-infected individuals is surprisingly large and may be found in more than 50% of some infected subpopulations. Most of these patients will not have overt renal disease, but there is a population of unknown size of patients with subclinical glomerular disease that has the potential to become clinically significant. In cases of hepatitis C virus-associated cryoglobulinemia, treatment remains focused on eradication of viremia, but interventions directed at B lymphocytes are increasingly utilized. The mechanisms of cryoglobulin formation and renal injury remain largely obscure, but recent evidence implicates the innate immune system in the initiation of disease. SUMMARY: The most common renal injury associated with hepatitis C virus infection, in patients both with and without evidence of cryoglobulinemia, is membranoproliferative glomerulonephritis. There has been increasing focus on defining the mechanisms that link these processes and the evolution of renal injury in all clinical settings of cryoglobulinemia. PMID- 18408475 TI - New aspects of the renin-angiotensin system: angiotensin-converting enzyme 2 - a potential target for treatment of hypertension and diabetic nephropathy. AB - PURPOSE OF REVIEW: Whereas angiotensin-converting enzyme promotes the formation of angiotensin II, angiotensin-converting enzyme 2 promotes the degradation of angiotensin II to angiotensin-(1-7). We review recent studies dealing with angiotensin-converting enzyme 2 in kidney disease and hypertension, and discuss the potential therapeutic benefit of increasing angiotensin-converting enzyme 2 activity in the treatment of these diseases. RECENT FINDINGS: In glomeruli from diabetic mice, angiotensin-converting enzyme 2 expression is downregulated, and pharmacological inhibition of angiotensin-converting enzyme 2 leads to worsening of albuminuria, increased mesangial matrix deposition and fibronectin expression. The deletion of the angiotensin-converting enzyme 2 gene in mice leads to worsening of angiotensin II-induced hypertension and has also been shown to cause glomerulosclerosis in aging male mice. SUMMARY: Angiotensin-converting enzyme 2 is a key enzyme in the renin-angiotensin system that favors the degradation of angiotensin I and angiotensin II. Angiotensin-converting enzyme 2 inhibition by pharmacological means and by genetic deletion worsens kidney disease in diabetic mice. Strategies geared to increasing angiotensin-converting enzyme 2 activity may provide a novel therapeutic target within the renin-angiotensin system by enhancing angiotensin II degradation that may complement the current approach of inhibiting angiotensin II formation and action. Amplifying angiotensin-converting enzyme 2 activity may have a potential therapeutic role for kidney disease and hypertension. PMID- 18408476 TI - Nephron number, glomerular volume, renal disease and hypertension. AB - PURPOSE OF REVIEW: To discuss studies evaluating associations of glomerular number (Nglom) and glomerular volume with hypertension and kidney disease. IMPORTANT FINDINGS: The association of low Nglom with hypertension and renal insufficiency was described in the 1930s. Many investigators have noted loss of glomeruli with age, with most disappearing entirely, and have proposed that hypertension follows. In a recent German study, hypertensive patients had fewer glomeruli and larger mean glomerular volumes than nonhypertensive people. Among the 10-fold range of Nglom in our multiracial autopsy series, the lowest were in Australian Aborigines, who have the highest rates of renal failure. Nglom fell with age. There was a five-fold range in mean glomerular volume and considerable heterogeneity in individual glomerular volumes within a patient. Larger mean glomerular volume and greater individual glomerular volume heterogeneity correlated with lower Nglom, larger body size, hypertension, and black race. Hypertension increased with age and was marked by glomerular enlargement, intimal thickening and higher rates of glomerulosclerosis. In whites and Aborigines, but not in US blacks, lower Nglom was associated with hypertension, while robust numbers were highly protective. SUMMARY: Higher mean glomerular volume and individual glomerular volume heterogeneity mark glomerular stress. Low Nglom is an important determinant of hypertension and renal disease. Many 'missing' nephrons have probably been lost during life, leaving little trace. Additional factors contribute to high rates of hypertension in blacks. PMID- 18408477 TI - Hypertensive nephrosclerosis. AB - PURPOSE OF REVIEW: Hypertensive nephrosclerosis is the second most common cause of end-stage renal disease, however morphologic evidence on the subject is poorly understood. A perennial and vexing problem in understanding kidney hypertension is that correlations between hypertension and vascular and glomerular lesions are only moderate, in part because all of these lesions are present to a greater or lesser degree in the normotensive, aging kidney, with racial differences in severity further compounding the problem. This review looks at newer data on this topic. RECENT FINDINGS: Recent data suggest that there are two different processes leading to glomerulosclerosis, and the combination of the two begins to explain why global correlations between hypertension and morphologic lesions are destined to remain poor. Arterial stiffening with increased pulse pressure down as far as the afferent arteriolar level likely plays an important role in the progression of glomerular lesions. Loss of renal autoregulation with glomerular hypertrophy, hyperfiltration, and focal segmental glomerulosclerosis is now recognized to contribute significantly to nephrosclerosis, particularly in the black population. Ischemic glomerulosclerosis, however, may ultimately be the most important lesion, with consequent hypoxia in the parenchyma beyond, leading to tubular atrophy and interstitial fibrosis. SUMMARY: Hypertensive nephrosclerosis should be seen as a process with two principal modes of glomerular sclerosis, ischemic and hypertrophic, with consequent focal segmental glomerulosclerosis, contributing variably to renal failure according to race and level of hypertension. PMID- 18408478 TI - The spectrum of focal segmental glomerulosclerosis: new insights. AB - PURPOSE OF REVIEW: Focal segmental glomerulosclerosis (FSGS) is a disease with diverse histologic patterns and etiologic associations. Genetic, toxic, infectious and inflammatory mediators have been identified. This review will focus on new evidence supporting the potential mechanistic basis underlying the histologic variants and their clinical relevance. RECENT FINDINGS: Evidence from animal models and in-vitro studies suggests that injury inherent within or directed to the podocyte is a central pathogenetic factor. Disruption of signaling from any of the podocyte's specialized membrane domains, including slit diaphragm, apical and basal membranes, or originating at the level of the actin cytoskeleton, may promote the characteristic response of foot process effacement. Irreversible podocyte stress leading to podocyte depletion through apoptosis or detachment is a critical mechanism in most forms of FSGS. In the collapsing variant, podocyte dysregulation leads to podocyte dedifferentiation and glomerular epithelial cell proliferation. SUMMARY: Translation studies in humans and new evidence from animal models have provided mechanistic insights into the diverse phenotypes of FSGS. PMID- 18408479 TI - Potential for peroxisome proliferator-activated receptor-gamma agonists in progression: beyond metabolism. AB - PURPOSE OF REVIEW: The thiazolidinediones are agonists for peroxisome proliferator-activated receptor-gamma, and promote insulin sensitization and improve dyslipidemia in patients with type II diabetes mellitus. Peroxisome proliferator-activated receptor-gamma transduces its actions by binding to common consensus response elements called peroxisome proliferator-activated receptor response elements, thus modifying expression of numerous genes. Peroxisome proliferator-activated receptor-gamma is widely expressed, including in macrophages and vascular smooth muscle cells. I will review recent novel insights into peroxisome proliferator-activated receptor-gamma agonist actions. RECENT FINDINGS: The thiazolidinediones have beneficial effects in humans with type II diabetes related in part to improvement of dysmetabolic syndrome, and also have beneficial effects on progressive renal damage in animal models of diabetic nephropathy and in models with severe hyperlipidemia. Peroxisome proliferator activated receptor-gamma agonists also have direct benefits on progressive renal injury independent of altering the dysmetabolic profile. SUMMARY: We will review selected evidence of actions of peroxisome proliferator-activated receptor-gamma agonists beyond metabolism, focusing on experimental work, and examine interactions with other key profibrotic mediators, including transforming growth factor-beta and the renin-angiotensin system. PMID- 18408480 TI - Relationship between chronic kidney disease prevalence and end-stage renal disease risk. AB - PURPOSE OF REVIEW: Incidence of end-stage renal disease has increased dramatically during the last 30 years and screening for early stages of chronic kidney disease is often suggested as a preventive measure. The relationship between chronic kidney disease and end-stage renal disease is complex, however, and recent studies have given some insights into this relationship. The review will summarize these studies and briefly discuss the clinical implications. RECENT FINDINGS: While the prevalence of chronic kidney disease is high in most Western countries, the incidence of end-stage renal disease differs substantially. The general increase in the incidence of end-stage renal disease seen in recent years may be partially explained by a lower cardiovascular mortality, allowing more patients with chronic kidney disease to develop end stage renal disease, and widening of entrance criteria for renal replacement therapy. Data do not, however, support these factors as explanatory for the existing international differences. These differences are better explained by different prevalences of diabetes and obesity as well as by differences in rate of progression from early chronic kidney disease stages to end-stage renal disease. Rate of progression seems to be affected by race, socioeconomic status and predialytic care. SUMMARY: Several mechanisms influence the relationship between chronic kidney disease and risk of end-stage renal disease. Decreased cardiovascular mortality and improved treatment availability may explain parts of the increase in the incidence of end-stage renal disease, and there are also large international differences in rates of progression from chronic kidney disease to end-stage renal disease that may be amendable by public health and predialytic care interventions. PMID- 18408481 TI - Cardiac disease in children with mild-to-moderate chronic kidney disease. AB - PURPOSE OF REVIEW: Studies in pediatric populations may provide sensitive insights into the prevalence, evolution and underlying mechanisms of cardiovascular complications of chronic kidney disease, without confounding factors such as aging and diabetes. This review reflects on recent echocardiographic studies assessing left-ventricular mass, geometry and function in children with mild-to-moderate chronic kidney disease. RECENT FINDINGS: A cross-sectional assessment of a large pediatric cohort established a 33% prevalence of left-ventricular hypertrophy in children with chronic kidney disease stages 2-4. Concentric and eccentric geometry are found equally frequently, suggesting variable contributions of enhanced left-ventricular preload and afterload and/or nonhemodynamic mechanisms. Concentric left ventricular hypertrophy is frequently associated with systolic dysfunction as evidenced by impaired circumferential shortening at the myocardial midwall level. Left-ventricular mass and dysfunction tend to be inversely correlated with glomerular filtration rate. Furthermore, male sex, obesity, anemia and inflammation appear to be associated with left-ventricular hypertrophy, whereas blood pressure shows surprisingly little association with left-ventricular mass, geometry or function. SUMMARY: Significant abnormalities of cardiac morphology and function are observed in children with early chronic kidney disease, demonstrating a marked intrinsic impact on the cardiovascular system. Regular echocardiographic screening appears indicated in the pediatric population with chronic kidney disease. PMID- 18408482 TI - Kidney disease in the elderly: update on recent literature. AB - PURPOSE OF REVIEW: We review recent literature on the epidemiology and outcomes of chronic kidney disease in the elderly and discuss implications for management. RECENT FINDINGS: Chronic kidney disease is common in the elderly and associated with substantial morbidity and mortality. The prognostic significance of a given level of estimated glomerular filtration rate, however, varies substantially by age. Estimates of the prevalence of chronic kidney disease at the population level and estimates glomerular filtration rate at the individual level are exquisitely sensitive to the methods used to arrive at these estimates. At present there is no clear consensus on the optimal approach to estimating glomerular filtration rate in elderly individuals in the clinical setting. Available evidence to guide management of chronic kidney disease is based largely on trials in younger individuals. It is unclear to what extent the results of these trials can be extrapolated to older individuals with chronic kidney disease. SUMMARY: We advise caution in applying current guidelines to the care of the large number of individuals aged 70 years and older with chronic kidney disease. Ideally, the care of these patients should be individualized and carefully integrated with the management of other comorbid conditions and with patient preferences. PMID- 18408483 TI - Nephrolithiasis as a systemic disorder. AB - PURPOSE OF REVIEW: Nephrolithiasis is a prominent public health issue. It imposes a substantial burden on human health and is a considerable financial expenditure for the nation. Numerous epidemiologic studies have shown a significant association between nephrolithiasis, obesity, hypertension and chronic kidney disease. The review highlights many of those emerging studies and sheds light on the importance of our recognition of kidney stones as a systemic illness. RECENT FINDINGS: Several cross-sectional retrospective studies have investigated the relationship between kidney stones and the metabolic syndrome. The various silent features of the metabolic syndrome, including type 2 diabetes, increased BMI, hypertension and dyslipidemia, are becoming progressively more recognized and independently associated with an increased risk of kidney stone formation. SUMMARY: Our further understanding of the underlying mechanisms in the connection between nephrolithiasis and the metabolic syndrome will stimulate the development of more effective preventive and therapeutic measures. PMID- 18408484 TI - Salt intake, blood pressure and clinical outcomes. AB - PURPOSE OF REVIEW: Salt intake has long been known to influence blood pressure among hypertensive patients, but its effect among those without overt hypertension, as well as its effects on cardiovascular disease itself, have been disputed. This report reviews the evidence for an effect on both blood pressure and cardiovascular disease, particularly among normotensive participants, including recent data from randomized trials. RECENT FINDINGS: The Dietary Approaches to Stop Hypertension study provided clear evidence for a short-term effect of sodium on blood pressure among those with and without hypertension. Longer term trials among nonhypertensive particpants suggest more modest effects of lifestyle sodium reduction interventions on blood pressure. The recent Trials of Hypertension Prevention follow-up study found that, despite small changes in blood pressure, the risk of cardiovascular disease was reduced by 25% among those in the sodium reduction intervention. A study of potassium salt substitution among Taiwanese veterans supports this finding. Sodium and potassium may act jointly in the development of hypertension and cardiovascular disease. SUMMARY: Average sodium consumption in the US population is excessively high, and well above recommended limits. Because most sodium derives from processed and restaurant foods, a reduction of sodium in these sources, as recently called for by the American Medical Association, is necessary to reduce exposure. PMID- 18408485 TI - Nephrogenic systemic fibrosis: epidemiology update. AB - PURPOSE OF REVIEW: The aim of this article is to outline the history of nephrogenic systemic fibrosis, a new and serious disease of patients with renal failure, and to give an update on its aetiology and prevalence. RECENT FINDINGS: Epidemiological and histochemical studies demonstrated that gadolinium-containing contrast agents used for magnetic resonance imaging have an essential causative role in most, if not all, cases of nephrogenic systemic fibrosis. One particular agent, gadodiamide, caused the majority of cases, but gadopentetate dimeglumine has also been implicated in several cases. Increasingly poor renal function, aberrations in calcium-phosphate metabolism and erythropoietin treatment seem to increase the risk of the disease and its severity. Up to 25-30% of patients with renal failure exposed to gadolinium-based contrast agents may develop nephrogenic systemic disease. The figure varies with type of gadolinium-based contrast agent and patient characteristics. There is no established curative treatment of the disease, but rapid recovery of renal function or hastened kidney transplantation may be beneficial. SUMMARY: It is mandatory to avoid exposing patients with renal failure to gadodiamide and gadopentetate dimeglumine. Other linear gadolinium based contrast agents should be used with great caution. It is premature to conclude whether cyclic gadolinium-based contrast agents are totally safe in renal failure. PMID- 18408486 TI - Estimating and measuring glomerular filtration rate in children. AB - PURPOSE OF REVIEW: Glomerular filtration rate is the best determinant of kidney function in children. A decline in glomerular filtration rate may be the only sign of renal disease. A knowledge of glomerular filtration rate allows the clinician to appropriately dose medications, administer fluids, and intervene early to prevent end-stage renal failure. The aim of this review is to discuss the current options for determining glomerular filtration rate in children and the latest advances in research on this topic. RECENT FINDINGS: Owing to the limited availability of inulin, the gold standard for measuring glomerular filtration rate, and the concern for using radioactivity, the plasma disappearance of iohexol has been utilized to measure glomerular filtration rate in children, and this represents a practical option for glomerular filtration rate determination in both clinical and research studies. By contrast, creatinine based estimates of glomerular filtration rate (e.g. Schwartz formula) are more practical for bedside use. To improve their precision, recent formulas have incorporated the use of additional endogenous markers, such as cystatin C, albumin, and blood urea nitrogen. SUMMARY: Iohexol plasma clearances should be used to accurately measure glomerular filtration rate in children. New estimating equations with multiple endogenous markers are being developed for everyday clinical use. However, at present, there is no substitute for an accurate glomerular filtration rate determination, particularly in children with conditions affecting muscle mass or body habitus. PMID- 18408487 TI - Bibliography. Current world literature. Renal immunology and pathology. PMID- 18408489 TI - Comparative effectiveness. PMID- 18408490 TI - Posterior segment complications after laser-assisted in situ keratomileusis. AB - PURPOSE OF REVIEW: The aim of this article is to review the retinal complications that may occur after laser-assisted in situ keratomileusis (LASIK). RECENT FINDINGS: One study investigated the incidence, characteristics, and surgical management of rhegmatogenous retinal detachment after LASIK surgery in myopia. The incidence of a rhegmatogenous retinal detachment after LASIK in this study was 0.033%. Another group reported a case of acute rhegmatogenous retinal detachment 14 h after LASIK surgery in a highly myopic patient. Two reports described a patient with early choroidal neovascular membrane after LASIK in a young myopic patient that was successfully managed by photodynamic therapy, and a patient with valsalva-like retinopathy following hyperopic LASIK. Another study evaluated the long-term effects of excimer laser treatment for ametropia after surgical treatment of rhegmatogenous retinal detachment with scleral buckles. A letter reported a case of a young man with an unusual grey ring around the optic disc and macular edema after LASIK-related corneal penetration and lens dislocation. Two case reports were on bilateral cystoid macular edema after phacoemulsification in post-LASIK eyes, and unilateral optic disc pit maculopathy after LASIK. A letter commented on the systemic contraindications for LASIK. SUMMARY: Serious complications after LASIK are infrequent. A dilated fundus examination is very important before LASIK and in every patient whose visual acuity after LASIK is not as good as expected. PMID- 18408491 TI - Update on treatments for diabetic macular edema. AB - PURPOSE OF REVIEW: Due to modest outcomes with macular laser, other treatment modalities for diabetic macular edema have been evaluated. Intravitreal triamcinolone acetonide, pars plana vitrectomy, oral protein kinase C inhibitors and, from more recently, anti-vascular endothelial growth factor therapy are reviewed. RECENT FINDINGS: Intravitreal triamcinolone acetonide may be more effective than laser. Intravitreal triamcinolone acetonide followed by laser may be more effective than intravitreal triamcinolone acetonide alone. Ruboxistaurin, a selective protein kinase C betainhibitor, reduced retinal vascular leakage in patients with diabetic macular edema and reduced the rate of sustained moderate visual loss in those with moderately severe to very severe non proliferative diabetic retinopathy. Several anti-vascular endothelial growth factor agents are under evaluation. Intravitreal pegaptanib sodium (0.3 mg) improved vision and reduced central retinal thickness compared to sham. Data on the other anti vascular endothelial growth factor agents is limited, but there are promising results, with ranibizumab (0.5 mg) and bevacizumab reducing foveal thickness and improving visual acuity in some patients with diabetic macular edema. SUMMARY: There remains no proven intervention that consistently prevents or reverses visual loss from diabetic macular edema in all patients. A variety of promising new medical and surgical therapies are under investigation, but further research is required to determine their role alone or in combination. PMID- 18408492 TI - Food and Drug Administration approval process for ophthalmic drugs in the US. AB - PURPOSE OF REVIEW: The purpose of this review is to provide the overall framework of the review and approval of ophthalmic drugs in the US. RECENT FINDINGS: Examples from the recent approval of the ranibizumab injection (Lucentis; Genentech, South San Francisco, California, USA) are offered to illustrate the current approval process at the Food and Drug Administration (FDA). SUMMARY: New drugs are brought to the US market after review of the New Drug Application or Biologic License Application by the FDA. This review process requires the teamwork of an interdisciplinary team of clinicians, scientists and regulatory personnel. This team is ultimately responsible for recommending to the Secretary of Health and Human Services (or designee) the approvability of a drug based on defined requirements including the drug's risk/benefit profile, quality and purity, and the drug's ability to be labeled effectively. Within the Center for Drug Evaluation and Research this review team for ophthalmic drugs resides primarily within the Division of Anti-Infective and Ophthalmology Products. PMID- 18408493 TI - The cost of vision for vitreoretinal interventions. AB - PURPOSE OF REVIEW: This article reviews the current status of treatments for age related macular degeneration from a cost-effectiveness perspective. RECENT FINDINGS: Treatments are now available to a broad range of patients with age related macular degeneration, but they are pharmaceutical based and require repeated evaluation and treatment. The resulting unprecedented levels of cost make cost-effectiveness considerations more prescient. Newer methods to evaluate cost-effectiveness and treatment outcomes have included utility and value-based analyses. These have attempted to utilize accurate, objective parameters, but are calibrated by subjective patient assessments and are limited by certain methodological assumptions. Treatment methods to improve outcomes are increasingly being considered within the context of cost-effectiveness. SUMMARY: Although improved understanding of basic, cellular processes has led to markedly improved treatment options for age-related macular degeneration within the past decade, the high costs and wide applicability threaten the balance and viability of the payor system. Accurate means of assessing cost-effectiveness are critical to defining clinical applications. PMID- 18408494 TI - The comparative effectiveness and cost-effectiveness of vitreoretinal interventions. AB - PURPOSE OF REVIEW: The comparative effectiveness of medical interventions has recently been emphasized in the literature, typically for interventions in a similar class. Value-based medicine, the practice of medicine based on the value (improvement in quality of life and/or length of life) conferred by medical interventions, allows a measure of comparative effectiveness of interventions across all of health care, no matter how disparate. This report discusses recent comparative effectiveness studies in the vitreoretinal literature. RECENT FINDINGS: Vitreoretinal interventions have good to excellent comparative effectiveness compared with commonly utilized interventions across health care, such as treatment for osteoporosis and hyperlipidemia. They also tend to be cost effective when an upper limit of $100 000/quality-adjusted life-year is utilized. SUMMARY: Value can be measured using either or both of two outcomes - the quality adjusted life-year gain and/or the percentage improvement in value - both of which allow for an evaluation of comparative effectiveness, which can be compared on the same scale for every intervention. This value can also be integrated with costs using the outcome of dollars expended per quality-adjusted life-year ($/quality-adjusted life-year, or the cost-utility ratio), which allows a comparison of cost-effectiveness across all interventions. The majority of vitreoretinal interventions confer considerable value and are cost-effective. PMID- 18408495 TI - Polypoidal choroidal vasculopathy and treatments. AB - PURPOSE OF REVIEW: This review assesses the current knowledge of the clinical characteristics of polypoidal choroidal vasculopathy and treatments. RECENT FINDINGS: Polypoidal choroidal vasculopathy is a disease with characteristic choroidal vascular abnormalities. Indocyanine green angiography is essential for diagnosis. The prevalence is higher in Asian people than in Caucasians. Photodynamic therapy is efficacious for treating polypoidal choroidal vasculopathy; 1-year results have shown greater benefit of photodynamic therapy than choroidal neovascularization secondary to age-related macular degeneration. Recurrence, however, seriously affects vision long term during follow-up after photodynamic therapy. The lower efficacy of bevacizumab- a full-length antibody of vascular endothelial growth factor- has been shown for polypoidal choroidal vasculopathy. SUMMARY: Although the polypoidal choroidal vasculopathy and age related macular degeneration have been known to share common genetic factors, its clinical characteristics including the different responses to photodynamic therapy suggest that polypoidal choroidal vasculopathy is a separate clinical entity from age-related macular degeneration. The results of photodynamic therapy for polypoidal choroidal vasculopathy are encouraging; however, recurrence may affect vision over time. Therapeutic modalities to inhibit development of the exudative choroidal vasculature of polypoidal choroidal vasculopathy are desirable. PMID- 18408496 TI - Ocular von Hippel-Lindau disease: clinical update and emerging treatments. AB - PURPOSE OF REVIEW: von Hippel-Lindau disease is an inherited, multisystemic cancer syndrome often involving the retina. This review will provide an update for the clinical characterization and treatment of ocular von Hippel-Lindau disease. RECENT FINDINGS: A comprehensive and quantitative clinical characterization of ocular von Hippel-Lindau disease has been limited by small patient numbers and nonrepresentative sampling. Recently, a large population of patients with clinically and genetically defined von Hippel-Lindau disease was systemically characterized in a single center, enabling a quantitative evaluation of the ocular involvement of this syndrome. Correlations between the nature of von Hippel-Lindau gene mutations and the ocular phenotype were also examined, providing clues as to how disruptions in von Hippel-Lindau protein function may result in eye disease. This understanding may be relevant to the development of new therapies targeting the molecular biology of von Hippel-Lindau disease, some of which are presently being investigated. SUMMARY: Quantitative studies enable a full characterization of the impact of von Hippel-Lindau disease on eye health and visual function. Establishing correlations between the genotype of the von Hippel-Lindau mutation and the phenotype of eye disease may inform us as to how ocular von Hippel-Lindau disease arises, and help guide molecular interventions in ocular von Hippel-Lindau disease. PMID- 18408497 TI - Diagnostic vitrectomy for the diagnosis and management of posterior uveitis of unknown etiology. AB - PURPOSE OF REVIEW: Diagnostic vitrectomy is performed in cases of posterior segment inflammation that are suspicious for malignancy or infection and require tissue confirmation. Advances in vitrectomy technique and laboratory tests have improved the diagnostic yield in these challenging cases. The methods for obtaining vitreous specimens and the diagnostic tests used to analyze them are discussed. RECENT FINDINGS: Ancillary tests have been invaluable in addressing the diagnostic limitations of traditional histopathologic and microbiologic analyses. Flow cytometry, gene rearrangement studies, and cytokine measurements are useful adjuncts to cytology for the diagnosis of malignancy, in particular primary intraocular lymphoma. Microbial DNA amplification by polymerase chain reaction and intraocular antibody measurement has been shown to detect the presence of infection by organisms that are difficult to culture. SUMMARY: When performed appropriately, diagnostic vitrectomy with carefully selected ancillary testing can lead to a definitive diagnosis in a large proportion of cases. PMID- 18408498 TI - Current trends in the management of intraocular foreign bodies. AB - PURPOSE OF REVIEW: The aim of this article is to present the recent literature on the preoperative assessment, intraoperative management and postoperative care of patients with intraocular foreign bodies and present a management algorithm. RECENT FINDINGS: The preoperative assessment includes an evaluation of concomitant, potentially life-threatening conditions, as well as a relevant history and ophthalmic examination. Neuroimaging, preferably with noncontrast helical computed tomography, provides excellent information about intraocular foreign body size, shape and location. The preoperative assessment provides valuable information for prognosis, counseling, and intraoperative guidance. The decision of delayed versus immediate intraocular foreign body removal must be guided by the patient's medical status, availability of adequate operating facilities and staff, and the presence of clinical endophthalmitis. The administration of perioperative systemic and topical third or fourth-generation fluoroquinolones may play a role in decreasing the incidence of postoperative endophthalmitis. Intravitreal antibiotics warrant consideration in patients with a Gram stain positive for organisms, in cases suspicious for endophthalmitis, and in high-risk settings. Both the material and size of the foreign body are considerations in the choice of instrument and extraction site used (sclerotomy versus scleral tunnel) at time of pars plana vitrectomy and intraocular foreign body removal. Postoperatively, patients should be monitored closely for the development of endophthalmitis, retinal detachment and proliferative vitreoretinopathy. SUMMARY: This review summarizes the recent literature on the assessment of intraocular foreign bodies and techniques for their removal. An algorithm provides guidelines for their perioperative and operative management. PMID- 18408499 TI - The treatment of branch retinal vein occlusion with bevacizumab. AB - PURPOSE OF REVIEW: New treatment modalities for branch retinal vein occlusion have recently been introduced. The role of intravitreal bevacizumab injections will be discussed and compared with laser photocoagulation and other novel intravitreal pharmacotherapies. RECENT FINDINGS: Argon laser photocoagulation is the single treatment for branch retinal vein occlusion that has been shown to reduce vision loss in a randomized controlled clinical trial. The effectiveness of this treatment is limited though. Currently, increasing data support the role of intravitreal bevacizumab as an effective treatment for patients with macular edema secondary to branch retinal vein occlusion. Multiple injections seem to be necessary in order to achieve visual stabilization, favorable and durable macular changes. The effect of a single injection seems to last 6-8 weeks. The most common treatment protocol is two to three injections over the first 5-6 months. Patients who had minimal or no response to laser therapy appeared to benefit from bevacizumab. No significant complications have been associated with its use but only short-term data are available. SUMMARY: Intravitreal bevacizumab appears to be a safe and effective treatment for macular edema associated with branch retinal vein occlusion, at least in the short term. Further randomized, controlled investigations are needed to assess long-term safety and efficacy of intravitreal bevacizumab. PMID- 18408500 TI - Posterior vitreous detachment and retinal detachment following cataract extraction. AB - PURPOSE OF THE REVIEW: Cataract surgery induces considerable vitreous modifications that can lead to posterior vitreous detachment. Studies indicate that these changes, whether combined or not with peripheral retinal degenerations such as lattice areas, carry risk for subsequent retinal breaks or detachment. RECENT FINDINGS: The incidence of retinal detachment increases after cataract extraction, but it decreases with improved surgical technique. Postoperative posterior vitreous detachment is a major promoter of retinal detachment after cataract surgery and is related to onset of most retinal tears leading to retinal detachment. Vitreous body destabilization increases the incidence of retinal detachment after surgery; this destabilization of the vitreous body can result from aggressive surgical technique, intraoperative or postoperative posterior capsule breaks, high myopia and retinal detachment in the fellow eye. These factors can act alone or together to determine the onset of anomalous forms of posterior vitreous detachment; in such cases, retinal breaks frequently develop and culminate in retinal detachment. SUMMARY: Vitreous modifications occurring after cataract extraction in patients without preoperative posterior vitreous detachment may be involved in the onset of postoperative retinal detachment. Careful postoperative follow-up is recommended in these patients to allow early detection of posterior vitreous detachment arising after surgery. PMID- 18408501 TI - Diabetic vitrectomy: the influence of lens status upon surgical outcomes. AB - PURPOSE OF REVIEW: Management of the lens in diabetic eyes undergoing vitrectomy has long been a source of controversy. Initially, the lens was removed during diabetic vitrectomy because of intraoperative changes. It was noted, however, that anterior segment neovascular complications were greater in aphakic eyes after diabetic vitrectomy, and subsequently the vitreoretinal surgeon attempted to spare the lens. Lens management in this regard continues to attract discussion. This report reviews recent trends in the management of the native lens in the diabetic eye undergoing vitrectomy. RECENT FINDINGS: The rate of cataract formation after diabetic vitrectomy is high in eyes left phakic. The rates of anterior segment neovascularization and retinal detachment after diabetic vitrectomy are similar in phakic and nonphakic eyes. The rate of subsequent reoperation after diabetic vitrectomy may be greater in eyes left phakic. SUMMARY: Although the management of the lens in an eye undergoing diabetic vitrectomy should be individualized, cataract extraction performed either before or in combination with vitrectomy may reduce the rate of subsequent reoperation. The vitreoretinal surgeon may consider rendering an eye nonphakic before or during diabetic vitrectomy to optimize outcomes. PMID- 18408502 TI - A clinical and optical coherence tomography study of choroidal colobomas. AB - PURPOSE OF REVIEW: This paper reviews current understanding of coloboma of the choroid, as evaluated clinically and using optical coherence tomography. RECENT FINDINGS: Studies of the margin of coloboma of the choroid with optical coherence tomography revealed several interesting features, including variability in transition from normal retina to intercalary membrane (gradual or abrupt), presence of subclinical retinal detachments, focal communications between the subretinal space and subintercalary membrane space in eyes with extra colobomatous retinal detachments, and inward humping of the eye wall in some cases. Pathological studies have stressed the importance of the intercalary membrane and the margin of coloboma as barriers for occurrence of retinal detachments. Vitrectomy and silicone oil tamponade could correct these retinal detachments in a majority of cases. SUMMARY: Optical coherence tomography can provide insight into pathology at the margin of the coloboma. Coupled with knowledge from histopathological studies, this information can guide the management of retinal detachments secondary to coloboma of the choroid with a high degree of success. PMID- 18408503 TI - Hereditary retinal disease. AB - PURPOSE OF REVIEW: This article reports recent advances in the diagnosis, genetic analysis, and treatment of hereditary retinal disease. RECENT FINDINGS: Clinicians and scientists continue to reveal the relationship between phenotype and genotype in hereditary retinal diseases. Persistent investigation and progressive technology are advancing the efficiency of mutation discovery. This technology is also leading to readily available genetic testing that aids clinicians in the diagnosis of these diseases. Functional genetic studies, and laboratory and human clinical trials are occurring that may lead to future treatment of these disorders. SUMMARY: A literature review of the recent discoveries and potential treatments for retinitis pigmentosa, Leber's congenital amaurosis, X-linked retinoschisis, Best's disease, Stargardt's disease, and congenital stationary night blindness is presented, along with a guide for clinicians seeking genetic testing of patients. PMID- 18408504 TI - Bibliography. Current world literature. Retinal, vitreous and macular disorders. PMID- 18408505 TI - Complicated things call for leaders in thought. PMID- 18408506 TI - Older antibiotic gains new respect as potent treatment for tuberculosis. Rifapentine is already approved for use with humans. PMID- 18408511 TI - Psychoeducation. Patient education gets respect. AB - Even those who are not psychiatric nurses find themselves caring for patients with psychiatric disorders. Adults with a mental health and/or substance abuse diagnosis accounted for 1 of 4 hospital stays in 2004. Whether the patient's problem is physical, mental, or both, most care is self-care. Psychoeducation improves health outcomes by optimizing self-care skills, engaging family and community supports, and promoting early recognition of problems and appropriate interventions. PMID- 18408513 TI - Moving beyond the anecdotal. Identifying the need for evidence-based research in hospice and palliative care. AB - The paucity of research in hospice and palliative care has resulted in the absence of a sound, scientific foundation for clinical practice. Most clinical practice performed in this area has not received systematic evaluation and therefore lacks scientific credibility. Many barriers exist, however, that present unique challenges for research with patients and families who have life limiting illness. Evaluation of key components for the successful performance of interdisciplinary research and integration in clinical settings are explored. PMID- 18408515 TI - Home-based chronic care. An expanded integrative model for home health professionals. AB - The Chronic Care Model (CCM) developed by is an influential and accepted guide for the care of patients with chronic disease. Wagner acknowledges a current healthcare focus on acute care needs that often circumvents chronic care coordination. He identifies the need for a "division of labor" to assist the primary care physician with this neglected function. This article posits that the role of chronic care coordination assistance and disease management fits within the purview of home healthcare and should be central to home health chronic care delivery. An expanded Home-Based Chronic Care Model (HBCCM) is described that builds on Wagner's model and integrates salient theories from fields beyond medicine. The expanded model maximizes the potential for disease self-management success and is intended to provide a foundation for home health's integral role in chronic disease management. PMID- 18408516 TI - Implementing a home telemonitoring program. AB - In the current healthcare arena, it is imperative that home healthcare agencies seek to reduce costs and produce positive outcomes from the care they give. Telemonitoring offers the ability to monitor closely the daily status of the patients that home healthcare agencies serve, to provide best practice care, and to produce positive outcomes from that care. This article presents the experience of how 1 hospital-based agency implemented a home telemonitoring program and describes the lessons learned throughout the process. The current healthcare arena demands cost reduction and improved patient care outcomes from home healthcare agencies (HHAs). The direction that each HHA chooses to take to answer these demands will set the stage for their future. This article describes the steps needed to implement a telemonitoring program. The description is based on how 1 HHA chose its course of action and set it in motion and shows what was learned throughout the process. PMID- 18408517 TI - Telenursing and home healthcare. the many facets of technology. AB - Telenursing was instituted as an effective mode for providing care to patients geographically distant from healthcare providers. Using telecommunications and information technology, nursing care is provided remotely to individuals. Nurses recognize the value of telecare and telehomecare as essential components of telenursing that give patients easy access to high-quality care and eliminate costs and difficulties associated with travel to healthcare facilities. Patient satisfaction with telenursing is related to prompt quality care from professional nurses. Telenursing continues to grow as a valuable method for providing nursing care, especially in home healthcare. PMID- 18408518 TI - When it's a hard act to swallow. Dysphagia in home care. PMID- 18408524 TI - VNAA Chronic Care Clearinghouse: help is a click away. PMID- 18408525 TI - Venlafaxine extended release versus citalopram in patients with depression unresponsive to a selective serotonin reuptake inhibitor. AB - Findings from the National Institute of Mental Health's Sequenced Treatment Alternatives to Relieve Depression trial indicate that approximately 50% of patients with major depressive disorder do not experience a treatment response after adequate first-line treatment with a selective serotonin reuptake inhibitor (SSRI). This study was designed to test the hypothesis that, after treatment failure with an SSRI, switching to venlafaxine extended release (ER) would offer advantages over switching to another SSRI, citalopram. The objectives of this trial were to compare the efficacy and safety of venlafaxine ER and citalopram in the treatment of moderate-to-severe depression in patients who did not experience a treatment response to an SSRI other than citalopram and to investigate the effects of severity of depression by categorizing treatment groups according to baseline severity. This was a 12-week, double-blind, randomized, parallel-group, multicenter study. Participants were adult outpatients who, following 8 weeks of monotherapy with an adequate dosing regimen of an SSRI other than citalopram and had not responded, met the diagnostic criteria for depression as described in the Diagnostic and statistical manual of mental disorders, fourth edition, and had a score > or =20 on the 21-item Hamilton Rating Scale for Depression (HAM-D21). After a 7-day screening period, patients were randomly assigned to receive venlafaxine ER 75 mg/day or citalopram 20 mg/day for the first 2 weeks. Doses could be increased every 2 weeks through week 6. Treatment lasted 12 weeks and was followed by a 1-week tapering period. Maximum dosages were venlafaxine ER 300 mg/day or citalopram 60 mg/day. The primary end point was the final on-therapy total HAM-D21 score. To investigate the treatment effects of the severity of depression on efficacy, a subgroup analysis was performed for baseline HAM-D21 total score < or =31 and >31. The analyses for HAM-D21, Montgomery-Asberg Depression Rating Scale (MADRS), Clinical Global Impressions - Severity (CGI-S), and Clinical Global Impressions - Improvement scores were based on intent-to treat (ITT) population, for both the primary analysis and subgroup analysis according to baseline HAM-D21 total scores < or =31 or >31. Safety assessments included the recording of adverse events (AEs). A total of 406 patients (200 venlafaxine ER, 206 citalopram) were randomly assigned and 396 patients were included in the ITT population (194 venlafaxine ER, 202 citalopram). Treatment groups were similar in terms of demographics and baseline psychiatric assessments. Two hundred and eighty-four patients (137 venlafaxine ER, 147 citalopram) were present in the ITT population with a baseline HAM-D21 total score < or =31 and 112 patients (57 venlafaxine ER, 55 citalopram) in the >31 group. In the primary analysis, there was no statistical difference between groups. The group with a baseline HAM-D21 total score of 20-31 did not differ significantly in any efficacy parameters. In the group with a baseline HAM-D21 total score >31, the venlafaxine ER group differed significantly from the citalopram group on the primary end point HAM-D21 total score (P=0.0121). The secondary end point CGI-S score was statistically significant (P=0.0359), although the MADRS total score (P=0.0930) was not. AEs were reported by 57.8 and 63.4% of venlafaxine ER and citalopram patients, respectively. Overall discontinuation rates were 24.5% for venlafaxine ER and 20.9% for citalopram. Discontinuation rates owing to an AE as a primary or secondary reason were 5.5% for venlafaxine ER and 5.3% for citalopram. Overall, venlafaxine ER and citalopram showed similar efficacy in patients who had an inadequate response to an SSRI. In the subset of more severely depressed patients, venlafaxine ER was significantly more effective on a number of efficacy measures. Patients who remain severely depressed following treatment with an SSRI may gain more benefit from the dual-action drug venlafaxine, rather than switching to another SSRI. PMID- 18408526 TI - The GRID-HAMD: standardization of the Hamilton Depression Rating Scale. AB - This report describes the GRID-Hamilton Depression Rating Scale (GRID-HAMD), an improved version of the Hamilton Depression Rating Scale that was developed through a broad-based international consensus process. The GRID-HAMD separates the frequency of the symptom from its intensity for most items, refines several problematic anchors, and integrates both a structured interview guide and consensus-derived conventions for all items. Usability was established in a small three-site sample of convenience, evaluating 29 outpatients, with most evaluators finding the scale easy to use. Test-retest (4-week) and interrater reliability were established in 34 adult outpatients with major depressive disorder, as part of an ongoing clinical trial. In a separate study, interrater reliability was found to be superior to the Guy version of the HAMD, and as good as the Structured Interview Guide for the Hamilton Depression Rating Scale (SIGH-D), across 30 interview pairs. Finally, using the SIGH-D as the criterion standard, the GRID-HAMD demonstrated high concurrent validity. Overall, these data suggest that the GRID-HAMD is an improvement over the original Guy version as well as the SIGH-D in its incorporation of innovative features and preservation of high reliability and validity. PMID- 18408527 TI - Weight gain during olanzapine treatment for psychotic depression: effects of dose and age. AB - Weight gain has often been associated with olanzapine treatment, yet little is known about the influence of patient age or cumulative dose on olanzapine associated weight gain. The first 118 participants in the National Institutes of Mental Health Study of the Pharmacotherapy of Psychotic Depression randomized clinical trial (ClinicalTrials.gov Registration NCT00056472) completing at least 4 weeks of treatment with olanzapine were analyzed to determine the relationship between weight gain, age, and cumulative olanzapine dose. Younger (age 18-59 years) and older (age 60+ years) participants received open-label olanzapine and either sertraline or placebo for up to 12 weeks. Linear mixed effect regression modeling was used to determine the effects of age and cumulative olanzapine dose on weight gain, controlling for potential confounders. Age was observed to have a significant negative association with weight gain (P=0.01), even after controlling for differences in cumulative dose and baseline body mass index. Each 10-year increase in age was associated with a decrease in mean weight gain over 12 weeks of approximately 0.6 kg (95% confidence interval: 0.14-1.05 kg). Cumulative olanzapine dose was also significantly associated with weight gain (P<0.0001). Approximately 60% of completers of the 12-week trial experienced clinically significant weight gain (> or =7% of baseline weight). PMID- 18408529 TI - Effects of antipsychotic medications on sleep in schizophrenia. AB - Schizophrenia is often accompanied by sleep problems. Evidence exists that these sleep difficulties have significant effects on individuals with this disorder. The mainstay of treatment for this condition is the administration of medications that have effects on neurotransmitter systems, which play an important role in sleep-wake function, including histamine, acetylcholine, serotonin, norepinephrine and dopamine. Little systematic attention, however, has been paid to how the sleep effects of these agents might play a role in the course of treatment, function and quality of life of schizophrenia patients. Schizophrenia medications can improve sleep problems and reverse the sleep architectural derangements that are common among patients with schizophrenia and, therefore, have the potential to improve the quality of life and functional capacity of the patient. Conversely, some sleep-wake effects of these medications can impair patient function and quality of life. In this study, we review the effects of schizophrenia medications and discuss their relevance to optimizing the clinical treatment of people with schizophrenia with regard to sleep-wake function. PMID- 18408530 TI - Clinical evidence for serotonin and norepinephrine reuptake inhibition of duloxetine. AB - Most antidepressants in clinical use are believed to function by enhancing neurotransmission of serotonin [5-hydroxytryptamine (5-HT)] and/or norepinephrine (NE) via inhibition of neurotransmitter reuptake. Agents that affect reuptake of both 5-HT and NE (serotonin-norepinephrine reuptake inhibitors) have been postulated to offer greater efficacy for the treatment of major depressive disorder (MDD). These dual-acting agents also display a broader spectrum of action, including efficacy for MDD and associated painful physical symptoms, diabetic peripheral neuropathic pain, generalized anxiety disorder, and fibromyalgia syndrome. Substantial preclinical evidence shows that duloxetine, an approved drug for the treatment of MDD, generalized anxiety disorder, and the management of diabetic peripheral neuropathic pain, inhibits reuptake of both 5 HT and NE. This paper reviews clinical and neurochemical evidence of duloxetine's effects on 5-HT and NE reuptake inhibition. The clinical evidence supporting duloxetine's effects on NE reuptake inhibition includes indirect measures such as altered excretion of NE metabolites, cardiovascular effects, and treatment emergent adverse event profiles similar to those for other drugs believed to act through the inhibition of NE reuptake. In summary, the data presented in this report provide clinical evidence of a mechanism for duloxetine involving both 5 HT and NE reuptake inhibition in humans and are consistent with preclinical evidence for 5-HT/NE reuptake inhibition. PMID- 18408528 TI - A 12-month, open-label, comparative study of quetiapine and risperidone in the acute and long-term treatment of schizophrenia. AB - This multicentre, observational, prospective, nonrandomized study compared the effectiveness and tolerability of quetiapine and risperidone in the acute and long-term treatment of schizophrenia in a clinical setting. Patients admitted to an acute unit with schizophrenia, schizophreniform or schizoaffective disorder (DSM-IV), who were prescribed quetiapine or risperidone (3 : 1 ratio) within the first week of treatment, according to the physician's usual practice, were recruited. In total, 492 patients (quetiapine: 367; risperidone: 125) were followed up at weeks 1 and 2, discharge and 6 and 12 months thereafter. Mean doses at 12 months were: quetiapine 718.5 mg/day and risperidone 7.0 mg/day. Efficacy measures (Brief Psychiatric Rating Scale, Clinical Global Impression Severity of Illness and Improvement) indicated similar results for both agents. No difference was found in rehospitalization rate with either drug. In terms of tolerability, orthostatic hypotension was more frequent with quetiapine, but extrapyramidal symptoms and male sexual dysfunction were more frequent with risperidone. In conclusion, quetiapine and risperidone had comparable effectiveness, but there were differences between treatments in their side effect profile. PMID- 18408532 TI - Children's Secrets. PMID- 18408531 TI - Efficacy and tolerability of second-generation antidepressants in social anxiety disorder. AB - A systematic review and meta-analysis were conducted to evaluate the comparative efficacy and tolerability of second-generation antidepressants in social anxiety disorder. Studies were identified by searching MEDLINE, Embase, The Cochrane Library, PsychLit, and the International Pharmaceutical Abstracts from January 1980 through October 2006. Comparative evidence was summarized and indirect comparisons were made using network meta-analysis. Only three head-to-head trials were identified; comparative trials found only minimal differences in efficacy between escitalopram and paroxetine, and no statistically significant differences in efficacy between extended-release venlafaxine and paroxetine. Pooled evidence from 15 placebo-controlled trials suggests that escitalopram [relative benefit (RB) 1.3; 95% confidence interval (CI) 1.2-1.5], paroxetine (RB 1.9; 95% CI 1.5 2.3), sertraline (RB 1.8; 95% CI 1.5-2.2), and venlafaxine (RB 1.7; 95% CI 1.5 1.9) all produce significantly more responders than placebo; evidence favored fluvoxamine over placebo but was not significant (RB 1.5; 95% CI 0.9-2.4). Network meta-analysis did not reveal differences in efficacy among drugs. Overall, a fair amount of evidence supports the efficacy of escitalopram, fluvoxamine, paroxetine, sertraline, and venlafaxine in social anxiety disorder. The drugs do not differ in efficacy, although their adverse event profiles do. PMID- 18408533 TI - Behavioral health screening in urban primary care settings: construct validity of the PSC-17. AB - The Pediatric Symptom Checklist-17 (PSC-17) is a brief form of the Pediatric Symptom Checklist that is designed to screen for behavioral health problems in primary care settings. It has been proposed to have three subscales: externalizing, internalizing, and attention problems. In the context of developing a behavioral health screening program in an inner-city primary care practice, we evaluated the construct validity of the PSC-17. A total of 331 families with children between 4 and 12 years of age who were seen for well-child care during the study were invited to complete the PSC-17 and 320 families (96.5%) did so. A confirmatory factor analysis was performed and the Comparative Fit Index and root mean square error of approximation fit statistics were calculated to determine whether the data fit the proposed three-factor model. We found that although the PSC-17 contained three subscales, several items did not load predominantly on the subscale that they were proposed to measure. Specifically, although the five items on the internalizing subscale loaded only on this subscale, only four of the seven externalizing items loaded exclusively on the externalizing subscale, and only two of the five attention items loaded exclusively on the attention problems subscale. Clinicians using the PSC-17 in urban low-income communities should recognize that the externalizing and attention problems subscales of the PSC-17 may not be valid measures of these dimensions of child behavior in this population. PMID- 18408534 TI - Development of self-awareness- III- Richard Hughes. PMID- 18408535 TI - Interdisciplinary behavioral intervention for life-threatening obesity in an adolescent with Prader-Willi syndrome - a case report. AB - OBJECTIVE: To demonstrate interdisciplinary behavioral rehabilitation for life threatening obesity in an adolescent female with Prader-Willi syndrome. METHODS: Body Mass Index (BMI), supplemental oxygen requirement, mobility, and tantrums were documented. A 600 kcal per day diet, physical therapy, and exercise were implemented in an inpatient setting. A behavioral program was employed by an interdisciplinary team to ensure dietary adherence and to increase patient compliance with therapy demands. Family caregivers were trained and continued the intervention at home. RESULTS: Important improvements were achieved and maintained including weight loss, decreased need for supplemental oxygen, greater exercise tolerance, and fewer tantrums. The patient's BMI decreased from 84.3 to 59.8 during inpatient treatment, and to 41.0 at last follow-up. CONCLUSIONS: Behavioral rehabilitation can be beneficial and potentially life saving for morbidly obese children with Prader-Willi syndrome, and perhaps for other obese youth with developmental or behavioral problems. PMID- 18408536 TI - Barking vocalizations and shaking movements in a 13-year old girl. AB - CASE: Erica is a 13-year old female who was hospitalized for a 4-week history of "barking" noises and 2 weeks of generalized shaking episodes. Four weeks prior to admission, she had a viral upper respiratory infection (URI) with cough which was treated with over-the-counter cough syrup. After resolution of the URI, she developed a persistent cough that turned into a "bark"-like vocalization. Both the mother and patient demonstrated the bark as an "arf" sound like that of a small dog at times, a large dog at others. These vocalizations were unrelenting, occurring 3-10 times per minute only while awake. They were not precipitated by any known factors nor were there alleviating factors. She could not voluntarily suppress the sound. In addition to the vocalizations, episodes of generalized shaking of the extremities began 2 weeks prior to admission. According to Erica's mother, each episode lasted about 10-60 seconds and occurred 30-40 times a day only when she was awake. These episodes were not rhythmic or symmetric, and they were not associated with bowel or bladder incontinence. There was no alteration of consciousness following the episodes. Erica denied any recollection of the barking or shaking.The medical evaluation did not reveal an etiology. It included a complete physical examination, a neurological examination, biochemical laboratory studies, and a negative video EEG study that captured 10 episodes of shaking. Child psychiatry was consulted. Erica was a pleasant, quiet female with slightly constricted affect and a normal speech pattern. She reported that she was a straight-A honors student who had difficulty trusting others; she said that she had no friends, only "associates." She said that she had periods of feeling "sad" and crying easily, but could not identify any recent stressful event. Episodes of barking and shaking diminished during the hospitalization. Erica was discharged home with outpatient psychiatric follow-up. PMID- 18408537 TI - Child soldiers: reintegration, pathways to recovery, and reflections from the field. PMID- 18408539 TI - An assessment of the walkability of two school neighborhoods in Greenville, North Carolina. AB - Walking to school provides the opportunity for increasing physical activity and for improving weight status in youth. Social ecological theory recognizes the link between supportive built environments and increased walking. To promote walking to school as a way to increase physical activity in youth, it is important to begin by assessing the presence and quality of sidewalks in school neighborhoods and then to advocate for improvements. The purpose of this study is to demonstrate the assessment of the walkability of two school neighborhood areas, using an evaluation process, which is designed for use by lay community members, that produces maps to disseminate the assessment findings to decision makers. A validated and reliable audit instrument was used to assess the walkability of 114 road segments in the immediately adjacent student enrollment areas surrounding two elementary schools. Ten variables characterizing the transportation and pedestrian environment were measured and used to calculate a walkability score for each road segment. Color-coded maps of the walkability scores for each road segment were created to display the patterns of walkability. Sidewalks were absent in 67 percent and 75 percent of the road segments surrounding the two schools, respectively. The maps reveal that the very few suitable roads for walking are isolated by networks of streets with no sidewalks. PMID- 18408540 TI - Public education strategies to increase awareness of stroke warning signs and the need to call 911. AB - Rapid identification and treatment of ischemic stroke can lead to improved patient outcomes. We implemented a 20-week public education campaign to increase community awareness of warning signs for stroke and the need to call 911. Telephone surveys were conducted in adults aged 45 years and older before and after the intervention to evaluate its impact. There was a significant increase in awareness of two or more warning signs for stroke from baseline to follow-up (67% to 83%). Awareness increased significantly among both men and women and younger and older respondents. There was no significant change in the proportion of respondents indicating that they would call 911 if they witnessed someone having a stroke (74% to 76%). However, after the campaign, an increased proportion of respondents indicated that they would call 911 if they experienced sudden speech problems (51% to 58%), numbness or loss of sensation (41% to 51%), or paralysis (46% to 59%) that would not go away. Our findings suggest that a high-intensity public education campaign can increase community awareness of the warning signs for stroke and the need to call 911. PMID- 18408542 TI - The built environment and physical activity: influencing physical activity through healthy design. PMID- 18408541 TI - The relationship between built park environments and physical activity in four park locations. AB - Despite widespread knowledge that physical activity is a valuable mechanism for preventing many lifestyle diseases, data from the 2001 Behavioral Risk Factor Surveillance System indicate that less than half of the US population met activity recommendations established by the Centers for Disease Control and Prevention. To increase physical activity levels, community officials around the United States have identified public parks as a convenient, low-cost resource to enable active living. However, the amenities of the built park environment that best facilitate active park visits are unknown. The current article describes the relationship of micro-level environmental components and park visitors' physical activity. Using the System for Observing Play and Recreation in Communities, park visitation patterns are documented and described according to user demographics. Broadly, visitors who were observed in park environments, which contained playgrounds, sport courts, and paths, were significantly more active than visitors in settings without these features. Furthermore, six types of built features were able to explain 58% of the variance in observed activity intensity among park visitors. Findings suggest that built features that support physical activity across the life span (paths and courts in particular) may be considered by community leaders seeking relatively low-cost mechanisms to promote physical activity among residents. PMID- 18408543 TI - Built environment and health: considerations from the field. PMID- 18408544 TI - Urban planning and public health: a story of separation and reconnection. PMID- 18408545 TI - Using policy and built environment interventions to improve public health. PMID- 18408546 TI - Interventions to increase interventions are needed. PMID- 18408547 TI - The surveys are in! The role of local government in supporting active community design. AB - BACKGROUND: Recent studies detailing how the built environment affects health have found that land use and community design may have an impact on reducing sedentary lifestyles and increasing physical activity. METHOD: However, there is little information available about the attitudes of local officials and professional staff who often are directly responsible for making decisions about land use and community design. RESULTS: This opportunistic study reports findings across five major surveys that investigated healthy community design knowledge, attitudes, and practice among local officials and professional staff. When possible, comparisons and contrasts of survey responses and policy implications are discussed. CONCLUSIONS: The sharing of these data across professions is an important step toward enhancing collaboration in the fields and in better understanding the needs of local leaders related to active living and healthy community design. PMID- 18408548 TI - Implementing smart growth strategies in rural America: development patterns that support public health goals. AB - Recent studies on obesity rates show alarming increases across the entire population. Some of these studies indicate higher rates of obesity in rural populations than urban and suburban populations. Obesity in children in rural places also outpaces their suburban and urban counterparts. Although a number of factors account for these differences, public health professionals and researchers have begun to recognize that conventional development patterns and land use policies in rural areas are playing an important role in the trend. Smart growth alternatives to current rural development patterns also support broad public health goals. Rural communities across America face a number of challenges, yet many are using smart growth development strategies to turn the challenges into opportunities. These strategies are structured in a way that builds on broadly held values in rural communities, ones that build upon the traditional development pattern and support multiple community goals. Public health professionals, managers, and academics will benefit from this discussion because it will explain the strategies that rural decision makers, planners, and citizens are adopting to create places that support multiple community goals including a built environment that sustains and promotes active living. PMID- 18408549 TI - Influencing design, promoting health. AB - This article provides details of the goals and accomplishments of the Land Use and Health Resource Team composed of public health officials, planners, researchers, extension agents, advocacy organizations, and the development community. The team seeks to understand local land use and health relationships, increase community engagement, and facilitate positive change in policies and the built environment. The team's action plan is (1) research local land use and health relationships; (2) undertake public education and community mobilization; (3) identify interventions, seek funding, and pilot tools to integrate health and planning; and (4) monitor outcomes. In 2005, the team produced a report presenting a picture of local conditions related to health and the built environment. Findings were unveiled at a stakeholder conference, and local best practices and future actions were discussed. A geographic information system based health impacts tool for use by planners in site plan review was developed. Funding was obtained to facilitate neighborhood organizations to complete self assessments and develop interventions related to community environments, physical activity, and healthy eating. The team achieved initial goals of creating partnerships and spurring awareness. Future activities include wider field testing of the health impacts tool, participation in a health-oriented master planning process, and monitoring change in health risk behaviors related to changes in the built environment. PMID- 18408550 TI - Creating tools for healthy development: case study of San Francisco's Eastern Neighborhoods Community Health Impact Assessment. AB - The San Francisco Department of Public Health recently completed a 2-year collaborative process, the Eastern Neighborhoods Community Health Impact Assessment (ENCHIA), to evaluate the potential positive and negative health impacts of land use development. ENCHIA resulted in a number of outcomes, including (1) a vision of a healthy San Francisco; (2) community health objectives to reflect the vision; (3) indicators and data to assess and measure the objectives and vision; (4) a menu of urban development strategies to advance the objectives; and (5) the Healthy Development Measurement Tool, an evidence based tool to support accountable, comprehensive, evidence-based, and health oriented planning. This case study describes the 18-month ENCHIA process, key outcomes, and lessons learned. The case study also provides an overview of the Healthy Development Measurement Tool and examples of its first applications to urban planning. Given the growing understanding of built environmental influences on health, ENCHIA illustrates the ability of a local public health agency to effectively engage in land use policy as a health promotion strategy. PMID- 18408551 TI - Designing healthier communities through the input of children. AB - Recently, the rapid growth in rates of diabetes, obesity, and hypertension has become significant headline news. Most of the discussion about reversing these trends has focused on promoting more exercise and healthier diets. Although encouraging, this is not the entire solution. Communities must realize the health impacts of a built environment where mixing land uses is illegal, homes and schools are located far from one another, and an automobile is necessary to get anywhere. This pattern of development significantly affects environmental quality and human health, especially for kids who must be driven everywhere. Including smart growth principles in local and regional land use decisions could help create communities that offer more opportunities for active living. This article explores student-led projects that developed new tools to assess the impacts of the built environment, and evaluates how effective these tools are in spurring collaboration among students, urban planners, and public healthcare practitioners. Specifically, this article provides researchers and practitioners in the public health field with information about student-based health assessment tools and their application. With this knowledge, researchers and practitioners would be better equipped to identify and capitalize on opportunities for smart growth and collaboration among public health advocates. The result would be healthier communities, a more informed populace, and the realization that how we plan our communities has a profound impact on how we manage our public health. PMID- 18408552 TI - Closing the grocery gap in underserved communities: the creation of the Pennsylvania Fresh Food Financing Initiative. AB - The issue of access to healthy foods has been central to the work of many community-based organizations around the country. One such organization, The Food Trust, launched an effective advocacy campaign to bring awareness and policy change to the issue. The Food Trust's efforts with its partners resulted in the creation of the Pennsylvania Fresh Food Financing Initiative, the nation's first statewide financing program to increase supermarket development in underserved areas. This article focuses on a key component of the advocacy campaign: the creation of an evidence-based report that served as a strong, credible foundation for the campaign. The steps that were taken to find partners, obtain and analyze the data, and disseminate the findings are described. In addition, the outcomes of the Fresh Food Financing Initiative are discussed. PMID- 18408553 TI - Factors influencing implementation of local policies to promote physical activity: a case study of Montgomery County, Maryland. AB - Policy interventions such as zoning ordinances, school facility sitting guidelines, capital improvement programs, and park master plans hold particular promise for promoting physical activity, especially at the local level. Despite increasing attention to the relationship between built environment characteristics and physical activity, there is a paucity of research on the extent to which local policies can promote or hinder physical activity. Furthermore, the impact of local policies on physical activity should depend on how effectively the policies are implemented. Based on the policy implementation literature and using Montgomery County, Maryland, as a case study, this study identifies factors related to the successful implementation of local policies hypothesized to influence physical activity. For our study, we conducted an extensive policy review and 17 in-depth interviews with 26 individuals. The interviews were transcribed, coded, and analyzed to identify the relevant factors that affect policy implementation. Our findings suggest that knowledge and awareness, commitment and capacity, intergovernmental coordination, the presence of an advocate or champion, and conflict influence physical activity policy implementation at the local level. Those trying to increase physical activity through policy could focus on these implementation features to help make policy implementation more successful. PMID- 18408554 TI - Aligning public health spending and priorities in Oklahoma. AB - All-state fiscal year 2005 public health-related spending in the state of Oklahoma was investigated including funds from federal, state, and local sources expended through the state health department and the two autonomous metropolitan health departments. METHODOLOGY: The cost finding and allocation methodology used a series of structured resource worksheets developed for this project that segregate public health department expenditures into six primary groups: disease and prevention; family health; community health; protective health; support and administrative services; and other. The six primary groups were further divided into 59 units and subunits. All financial data were provided directly by staff in the public health agencies working closely with project staff. The data were analyzed along three lines: (1) level of health department (state, metro, other local); (2) revenue source (federal, state, local); and (3) public health function (behaviors, health conditions, direct services, population health). RESULTS AND CONCLUSIONS: Public health officials may not have necessary information on the multiple sources and applications of revenue, categories of expense, operational control of resources, and the inherent restrictions upon the use of those resources. The study gave the city-county and state health officials a new and more complete picture of public health spending in Oklahoma, which catalyzed a dialogue between the commissioner and the directors to explore ways for local priorities to be incorporated into the direct state spending. PMID- 18408555 TI - Pesticide exposure surveillance and prevention skills of staff in eastern North Carolina health departments. AB - BACKGROUND: In 2006, the NC Division of Public Health reported on the prenatal occupational pesticide exposures of three female migrant farmworkers whose newborns had birth defects. Unknown response capabilities from local health departments to pesticide exposures led to this pilot study to determine the skills of staff in eastern NC health departments regarding pesticide exposure surveillance and prevention. METHODS: Randomly selected staff (n = 193) from seven health departments participated in the study. Using a self-administered questionnaire, data were gathered on staff's pesticide education, knowledge of resources to assist with pesticide issues, frequency of pesticide surveillance/education activities linked to routine services, and pesticide exposure prevention behaviors that were screened/educated for. FINDINGS: Twelve participants were environmental health staff. All staff completed an average of less than 0.17 pesticide education courses, have minimal knowledge of resources, and conduct limited pesticide exposure surveillance/education. Pesticide exposure surveillance and prevention activities were reported by less than 30 percent of all staff. CONCLUSIONS: Inadequately trained staff lack access to, or knowledge of, resources to effectively engage in pesticide exposure surveillance and prevention activities. RECOMMENDATIONS: To enhance information on surveillance and epidemiology of pesticide chronic exposures at the NC Division of Public Health and to conduct further data analyses of this study. PMID- 18408556 TI - The role of local public health agencies in pesticide exposure. PMID- 18408557 TI - Zoning, urban planning, and the public health practitioner. PMID- 18408558 TI - Managing the difficult conversation. PMID- 18408559 TI - Translating the cost benefits of the formal and systematic integration of public health in community design. AB - The National Association of County and City Health Officials (NACCHO) is the national organization representing local health departments. NACCHO supports efforts that protect and improve the health of all people and all communities by promoting national policy, developing resources and programs, seeking health equity, and supporting effective local public health practice and systems. PMID- 18408561 TI - MDR1 diplotypes as prognostic markers in multiple myeloma. AB - OBJECTIVE: The aim of this study was to evaluate the effect of diplotypes of MDR1 polymorphisms at positions 2677 and 3435 on the clinical outcome of multiple myeloma, in terms of response to the therapy and overall survival (OS). METHODS: G2677T/A SNP was analysed by RFLP-PCR assay on 110 patients, treated with dexamethasone, doxorubicin (adryamicin) and vincristine regimen, followed by autologous stem cell transplantation. RESULTS: Treatment efficacy was not related to G2677T/A SNP, whereas the OS of G/G carriers was significantly shorter than that of T/T or G/T patients. Similar results were previously reported for MDR1 C3435T polymorphism. Given that these two single nucleotide polymorphisms are in strong linkage disequilibrium, we analyzed the effects of the most frequent haplo/diplotypes and the survival probability was lower for GC/GC patients (55%) than for GC/TT and TT/TT carriers (>80%; log-rank test, P=0.03). Interestingly, the effect of MDR1 diplotype on the OS seems to be confined to autologous stem cell transplantation nonresponders. CONCLUSION: These results support the hypothesis that genetic variability of MDR1 should be considered as an important factor that influences the clinical outcome of multiple myeloma. PMID- 18408560 TI - Association of corticotropin-releasing hormone receptor-2 genetic variants with acute bronchodilator response in asthma. AB - OBJECTIVE: Corticotropin-releasing hormone receptor (CRHR)-2 participates in smooth muscle relaxation response and may influence acute airway bronchodilator response to short-acting beta2-agonist treatment of asthma. We aim to assess associations between genetic variants of CRHR2 and acute bronchodilator response in asthma. METHODS: We investigated 28 single nucleotide polymorphisms in CRHR2 for associations with acute bronchodilator response to albuterol in 607 Caucasian asthmatic patients recruited as part of the Childhood Asthma Management Program. Replication was conducted in two Caucasian adult asthma cohorts--a cohort of 427 participants enrolled in a completed clinical trial conducted by Sepracor Inc. (Massachusetts, USA) and a cohort of 152 participants enrolled in the Clinical Trial of Low-Dose Theophylline and Montelukast conducted by the American Lung Association Asthma Clinical Research Centers. RESULTS: Five variants were significantly associated with acute bronchodilator response in at least one cohort (PT, 2677G>T/A and 3435C>T were assayed in the absence and presence of known inhibitors and AEDs. The inhibitory potency of the tested drugs from the dose-response relationships was cyclosporin A>verapamil> phenytoin> carbamazepine> lamotrigine>phenobarbital>valproic acid, levetiracetam, gabapentin. The silent polymorphisms combination (1236T-3435T) and triple haplotypes (1236T-2677A/T-3435T) resulted in profoundly less effective inhibition against substrates with significantly lower intracellular substrate concentration. These results confirmed that ABCB1 polymorphisms were associated with clinical responses of AEDs. CONCLUSION: Our findings demonstrated that human ABCB1 polymorphisms may alter the interactions between Pgp and substrates, and provided functional evidence for ABCB1 haplotypes-associated epilepsy treatment responses. PMID- 18408563 TI - Stargazin involvement with bipolar disorder and response to lithium treatment. AB - OBJECTIVES: Multiple reports have implicated chromosomal region 22q13.1 in both schizophrenia and bipolar disorder. The calcium channel gamma-2 subunit gene (cacng2, Stargazin) located on 22q13.1 was recently reported to be associated with schizophrenia. We aimed to examine the expression levels of Stargazin in post-mortem brain samples of patients with schizophrenia, patients with bipolar disorder (BPD) and healthy controls, test for genetic association between Stargazin and these disorders and test for genetic association between Stargazin and response to lithium treatment. METHODS: Expression analysis was carried out by quantitative reverse transcription-PCR in RNA samples from dorsolateral prefrontal cortices of patients with schizophrenia, patients with BPD and controls (n=35 each). Twelve single nucleotide polymorphisms encompassing Stargazin were genotyped in DNA samples from two cohorts, 'Aberdeen' and 'Cagliari' (n=410, 170, respectively). Patients were treated with lithium and divided into groups according to their response. RESULTS: A 1.6-fold overexpression of Stargazin was observed in patients with BPD (P=0.000036). No difference in expression was observed in patients with schizophrenia. None of the 12 genotyped single nucleotide polymorphisms were associated with BPD, but three of them were significantly associated with lithium response: one in both cohorts (rs2284017) and two (rs2284018, rs5750285) each in a different cohort. Haplotype analysis revealed significant 'response-protective' and 'response-inhibitive' haplotypes in both cohorts. CONCLUSION: Our findings suggest that Stargazin dysregulation may be involved with the pathophysiology of BPD, but not with that of schizophrenia, and that Stargazin polymorphisms may play a role in the response to lithium treatment. PMID- 18408564 TI - Impact of MDR1 and CYP3A5 on the oral clearance of tacrolimus and tacrolimus related renal dysfunction in adult living-donor liver transplant patients. AB - OBJECTIVE: The potential influence of the multidrug resistance 1 (MDR1) gene and the cytochrome P450 (CYP) genes, CYP3A4 and CYP3A5, on the oral clearance (CL/F) of tacrolimus in adult living-donor liver transplant patients was examined. Furthermore, the development of renal dysfunction was analyzed in relation to the CYP3A5 genotype. METHODS: Sixty de novo adult liver transplant patients receiving tacrolimus were enrolled in this study. The effects of various covariates (including intestinal and hepatic mRNA levels of MDR1 and CYP3A4, measured in each tissue taken at the time of transplantation, and the CYP3A5*3 polymorphism) on CL/F during the first 50 days after surgery were investigated with the nonlinear mixed-effects modeling program. RESULTS: CL/F increased linearly until postoperative day 14, and thereafter reached a steady state. The initial CL/F immediately after liver transplantation was significantly affected by the intestinal MDR1 mRNA level (P<0.005). Furthermore, patients carrying the CYP3A5*1 allele in the native intestine, but not in the graft liver, showed a 1.47 times higher (95% confidence interval, 1.17-1.77 times, P<0.005) recovery of CL/F with time than patients having the intestinal CYP3A5*3/*3 genotype. The cumulative incidence of renal dysfunction within 1 year after transplantation, evaluated by the Kaplan-Meier method, was significantly associated with the recipient's but not donor's CYP3A5 genotype (*1/*1 and *1/*3 vs. *3/*3: recipient, 17 vs. 46%, P<0.05; donor, 35 vs. 38%, P=0.81). CONCLUSION: These findings suggest that the CYP3A5*1 genotype as well as the MDR1 mRNA level in enterocytes contributes to interindividual variation in the CL/F of tacrolimus in adult recipients early after living-donor liver transplantation. Furthermore, CYP3A5 in the kidney may play a protective role in the development of tacrolimus-related nephrotoxicity. PMID- 18408565 TI - The effect of SLCO1B1*15 on the disposition of pravastatin and pitavastatin is substrate dependent: the contribution of transporting activity changes by SLCO1B1*15. AB - OBJECTIVE: This study was addressed to understand the underlying mechanism of the substrate-dependent effect of genetic variation in SLCO1B1, which encodes OATP1B1 (organic anion transporting polypeptide) transporter, on the disposition of two OATP1B1 substrates, pravastatin and pitavastatin, in relation to their transport activities. METHODS: The uptake of pravastatin, pitavastatin, and fluvastatin was measured in oocytes overexpressing SLCO1B1*1a and SLCO1B1*15 to compare the alterations of in-vitro transporting activity. After 40-mg pravastatin or 4-mg pitavastatin was administered to 11 healthy volunteers with homozygous genotypes of SLCO1B1*1a/*1a and SLCO1B1*15/*15, the pharmacokinetic parameters of pravastatin and pitavastatin were compared among participants with SLCO1B1*1a/*1a and SLCO1B1*15/*15 genotypes. RESULTS: The uptake of pravastatin and pitavastatin in SLCO1B1*15 overexpressing oocytes was decreased compared with that in SLCO1B1*15, but no change occurred with fluvastatin. The fold change of in-vitro intrinsic clearance (Clint) for pitavastatin in SLCO1B1*15 compared with SLCO1B1*1a was larger than that of pravastatin (P<0.0001). The clearance (Cl/F) of pitavastatin was decreased to a greater degree in participant with SLCO1B1*15/*15 compared with that of pravastatin in vivo (P<0.01), consistent with in-vitro study. As a result, Cmax and area under the plasma concentration time curve of these nonmetabolized substrates were increased by SLCO1B1*15 variant. The greater decrease in the transport activity for pitavastatin in SLCO1B1*15 variant compared with SLCO1B1*1a was, however, associated with the greater effect on the pharmacokinetics of pitavastatin compared with pravastatin in relation to the SLCO1B1 genetic polymorphism. CONCLUSION: This study suggests that substrate dependency in the consequences of the SLCO1B1*15 variant could modulate the effect of SLCO1B1 polymorphism on the disposition of pitavastatin and pravastatin. PMID- 18408566 TI - Trinucleotide repeat variants in the promoter of the thiopurine S methyltransferase gene of patients exhibiting ultra-high enzyme activity. AB - Thiopurine S-methyl transferase (TPMT) is a cytosolic enzyme that catalyses the S methylation of the thiopurine immunosuppressants. To date, 22 variants have been identified that are predictive of decreased TPMT activity. In contrast, no molecular explanation has been found for the 1-2% of Caucasians who exhibit ultra high TPMT activity. Here, we report the characterization of polymorphisms within a trinucleotide (GCC) repeat element of the TPMT promoter in two patients with inflammatory bowel disease exhibiting the highest TPMT activity from two testing centres. The first patient was heterozygous for a variant allele carrying seven GCC repeats [(GCC)7], whereas the second patient was heterozygous for a variant allele containing five GCC repeats [(GCC)5]. Fifty patients with inflammatory bowel disease with normal TPMT activity were all homozygous for six GCC repeats [(GCC)6]. Of 200 healthy controls, five were found to be heterozygous for the (GCC)7 variant. Within in vitro reporter gene assays, the mean luciferase activities of the (GCC)6, (GCC)7, and (GCC)5 constructs were 8.0+/-0.26, 13.2+/ 0.10 and 12.3+/-0.12, respectively. The significant increase in activity observed for (GCC)5 and (GCC)7 compared with (GCC)6 (P-value A variant was reduced in comparison with the wild-type control. Intestinal biopsy samples revealed that BCRP protein and mRNA expression did not significantly differ between patients with 34GG/421CC versus patients with 34GG/421CA genotypes. Remarkably, in subjects with 34GG/421CA genotype, sulfasalazine area under the concentration-time curve was 2.4-fold greater compared with 34GG/421CC subjects (P<0.05). This study links commonly occurring SNPs in BCRP with significantly increased oral sulfasalazine plasma exposure in humans. Accordingly, sulfasalazine may prove to have utility as in vivo probe for assessing the clinical impact of BCRP for the disposition and efficacy of drugs. PMID- 18408569 TI - Transplant recipient's knowledge of posttransplant malignancy risk: implications for educational programs. PMID- 18408570 TI - "Take another little piece of my heart now": should endomyocardial biopsy remain the gold standard? PMID- 18408571 TI - Declining outcomes in simultaneous liver-kidney transplantation in the MELD era: ineffective usage of renal allografts. AB - BACKGROUND: When the United Network for Organ Sharing changed its algorithm for liver allocation to the model for end-stage liver disease (MELD) system in 2002, highest priority shifted to patients with renal insufficiency as a major component of their end-stage liver disease. An unintended consequence of the new system was a rapid increase in the number of simultaneous liver-kidney transplants (SLK) being performed yearly. METHODS: Adult recipients of deceased donor liver transplants (LT, n=19,137), kidney transplants (n=33,712), and SLK transplants (n=1,032) between 1987 and 2006 were evaluated based on United Network for Organ Sharing data. Recipients were stratified by donor subgroup, MELD score, pre- versus post-MELD era, and length of time on dialysis. Matched control analyses were performed, and graft and patient survival were analyzed by Kaplan-Meier and Cox proportional hazards analyses. RESULTS: MELD era outcomes demonstrate a decline in patient survival after SLK. Using matched-control analysis, we are unable to demonstrate a benefit in the SLK cohort compared with LT, despite the fact that higher quality allografts are being used for SLK. Subgroup analysis of the SLK cohort did demonstrate an increase in overall 1-year patient and liver graft survival only in those patients on long-term dialysis (> or =3 months) compared with LT (84.5% vs. 70.8%, P=0.008; hazards ratio 0.57 [95% CI 0.34, 0.95], P=0.03). CONCLUSION: These findings suggest that SLK may be overused in the MELD era and that current prioritization of kidney grafts to those liver failure patients results in wasting of limited resources. PMID- 18408572 TI - IL-17 production elicited by allo-major histocompatibility complex class II recognition depends on CD25posCD4pos T cells. AB - BACKGROUND: Interleukin (IL)-17 is involved in autoimmune inflammatory disorders and naturally occurring CD25pos regulatory T cells were shown to promote IL-17 synthesis. Because IL-17 is overproduced in certain types of allograft rejection, it is important to characterize the cells responsible for IL-17 synthesis and to define how IL-17 is regulated during alloimmune responses. METHODS: Splenic CD4pos T cells were isolated from C57BL/6 mice and fractionated according to CD25 expression. T cells were stimulated by major histocompatibility complex class II mismatched bone marrow-derived dendritic cells from bm12 mice, either immature or made mature by exposure to lipopolysaccharide. To track T cell populations, CD25negCD4pos and CD25posCD4pos were isolated from Thy1.1 and congenic Thy1.2 mice, respectively. Cell proliferation was quantified by CFSE dilution. IL-17 producing cells and FOXP3pos cells were enumerated by intracytoplasmic staining and cytokine levels in culture supernatants were measured by ELISA. RESULTS: Addition of CD25posCD4pos T cells to CD25negCD4pos T cells inhibited IL-2, interferon-[gamma], and IL-13 production but promoted IL-17 synthesis on stimulation by allogenic immature DC. In this setting, IL-17 originated from CD25intCD4posFOXP3neg memory T cells, which depend on IL-2 to produce IL-17. Alloreactive CD25negCD4pos T cells were also induced to produce IL-17 when stimulated by mature DC in the presence of CD25highCD4posFOXP3pos T cells. CONCLUSIONS: We conclude that (1) the cellular source of IL-17 during an antiallo major histocompatibility complex class II response depends on the maturation status of allogenic DC, (2) whereas suppressing Th1 and Th2 cytokine synthesis, naturally occurring regulatory T cells, allow IL-17 production by alloreactive CD4pos T cells. PMID- 18408573 TI - Donor and isolation variables predicting human islet isolation success. AB - BACKGROUND: Recent advances in the fields of islet transplantation and in vitro islet cell expansion place a renewed emphasis on the optimization of islet isolation from cadaveric human donor organs. We retrospectively analyzed 171 islet isolations to identify variables that predict islet yield and isolation success. METHODS: Cadaveric human donor pancreata were procured and processed according to established protocols. Donor-, procurement-, and isolation-related variables were analyzed for correlation with islet yield and isolation success (> or =250,000 islet equivalents). RESULTS: Univariate analysis suggested correlations between islet yield and donor age (P<0.005), body surface area (P<0.005), duration of enzymatic digestion (P<0.001), and pancreatic beta-cell volume (P<0.05). Donor sex (P<0.01), procurement team (P<0.05), and peridigestion serine protease inhibition (P<0.05) affected islet yield, whereas enzyme lot (P<0.01) and pancreatic fatty infiltration (P<0.05) influenced isolation success. By logistic regression, donor sex and age, and duration of enzymatic digestion could predict a successful isolation with 72% accuracy. The use of Liberase CI improved islet yield (P<0.05) in young donors (< or =25 years). CONCLUSIONS: While donor-related variables are useful in predicting islet yield, these are likely surrogates for pancreatic beta-cell volume. Enzyme lot, and the associated duration of enzymatic digestion (P<0.05), appears to be key determinants of isolation success. PMID- 18408574 TI - Coagulation defects do not predict blood product requirements during liver transplantation. AB - BACKGROUND: In our experience, correction of coagulation defects with plasma transfusion does not decrease the need for intraoperative red blood cell (RBC) transfusions during liver transplantation. On the contrary, it leads to a hypervolemic state that result in increased blood loss. A previous study has shown that plasma transfusion has been associated with a decreased 1-year survival rate. The aim of this prospective study was to evaluate whether anesthesiologists could reduce RBC transfusion requirements during liver transplantation by eliminating plasma transfusion. METHODS: Two hundred consecutive liver transplantations were prospectively studied over a 3-year period. Patients were divided into two groups: low starting international normalized ratio (INR) value <1.5 and high INR > or =1.5. Low central venous pressure was maintained in all patients before the anhepatic phase. Coagulation parameters were not corrected preoperatively or intraoperatively in the absence of uncontrollable bleeding. Phlebotomy and auto transfusion of blood salvaged were used following our protocol. Independent variables were analyzed in both univariate and multivariate fashion to find a link with RBC transfusions or decreased survival rate. RESULTS: The mean number of intraoperative RBC units transfused was 0.3+/-0.8. Plasma, platelet, albumin, and cryoprecipitate were not transfused. In 81.5% of the patients, no blood product was used during their transplantation. The average final hemoglobin (Hb) value was 91.2+/-15.0 g/L. There were no differences in transfusional rate, final Hb, or bleeding between two groups (low or high INR values). The overall 1-year survival rate was 85.6%. Logistic regression showed that avoidance of plasma transfusion, phlebotomy, and starting Hb value were significantly linked to liver transplantation without RBC transfusion. The need for intraoperative RBC transfusion and Pugh's score were linked to the decreased 1-year survival rate. CONCLUSION: The avoidance of plasma transfusion was associated with a decrease in RBC transfusions during liver transplantation. There was no link between coagulation defects and bleeding or RBC or plasma transfusions. Previous reports indicating that it is neither useful nor necessary to correct coagulation defects with plasma transfusion before liver transplantation seem further corroborated by this study. We believe that this work also supports the practice of lowering central venous pressure with phlebotomy to reduce blood loss, during liver dissection, without any deleterious effect. PMID- 18408575 TI - Lipoprotein-associated phospholipase A2 predicts progression of cardiac allograft vasculopathy and increased risk of cardiovascular events in heart transplant patients. AB - BACKGROUND: Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a risk factor for coronary artery disease (CAD) in nontransplant patients. We evaluated the association between Lp-PLA2, cardiac allograft vasculopathy (CAV) assessed by 3D intravascular ultrasound, and incidence of cardiac adverse events in heart transplant recipients. MATERIALS AND METHODS: Fasting blood samples were obtained and stored from a cross-section of 112 cardiac transplant recipients attending the Mayo cardiac transplant clinic in 2000 to 2001, mean of 4.7 years after transplant. Lp-PLA2 was measured in plasma aliquots using an enzyme-linked immunoassay. Fifty-six of these patients subsequently underwent two 3D intravascular ultrasound studies in 2004 to 2006 12 months apart. Cardiovascular (CV) events included percutaneous coronary intervention, coronary artery bypass grafting (CABG), reduction in left ventricular ejection fraction (LVEF) < or =45% secondary to CAV and CV death. RESULTS: High Lp-PLA2 level was associated with increase in plaque volume (r=0.43, P=0.0026) and percent plaque volume (r=0.45, P=0.0004). The association remained significant after adjusting for clinical and lipid variables. During follow-up of 5.1+/-1.6 years, 24 CV adverse events occurred in 15 of 112 (13%) heart transplant patients. Lp-PLA2 level>236 ng/mL (higher tertile) identified a subgroup of patients having a 2.4-fold increase of relative risk for combined endpoint of CV events (percutaneous coronary intervention, CABG, LVEF<45%, and CV death; 95% CI 1.16-5.19, P=0.012) compared with patients with Lp-PLA2< or =236 ng/mL. CONCLUSIONS: Lp-PLA2 is independently associated with progression of CAV and predicts a higher incidence of CV events and CV death in transplant patients. This finding supports the concept that systemic inflammation is an important mediator of CAV. Lp-PLA2 may be a useful marker for risk of CAV and a therapeutic target in posttransplant patients. PMID- 18408576 TI - Limited utility of endomyocardial biopsy in the first year after heart transplantation. AB - BACKGROUND: Surveillance endomyocardial biopsies (EMBs) are used for the early diagnosis of acute cardiac allograft rejection. Protocols became standardized in an earlier era and their utility with contemporary immunosuppression has not been investigated. METHODS: We studied 258 patients after orthotopic heart transplantation comparing 135 patients immunosuppressed by mycophenolate mofetil (MMF) with 123 patients treated by azathioprine (AZA); both with cyclosporine and corticosteroids after induction therapy with rabbit antithymocyte globulin. Fifteen EMBs were scheduled in the first year. Additional EMBs were performed for suspected rejection, after treatment, or for inadequate samples. The MMF group had 1875 EMBs vs. 1854 in the AZA group. RESULTS: The yield of International Society for Heart and Lung Transplantation (ISHLT) grade> or =3A biopsy-proven acute rejection (BPAR) was 1.87% per biopsy (35 of 1875) with MMF vs. 3.13% (58 of 1854) with AZA P=0.024. The number of clinically silent BPAR ISHLT grade > or =3A (the true yield of surveillance EMBs) was 1.39% (26 of 1875) of biopsies MMF vs. 2.1% (39 of 1854) AZA, P=0.48. There were five serious complications requiring intervention or causing long-term sequelae; 0.13% (5 of 3729) per biopsy and 1.94% (5 of 258) per patient. The incidence of all definite and potential complications was 1.42% (53 of 3729) per biopsy and 20.5% (53 of 258) per patient. There was no biopsy-related mortality. CONCLUSION: The yield of BPAR was low in the AZA group and very low in the MMF group. The incidence of complications was also low, but repeated biopsies led to a higher rate per patient. Routine surveillance EMBs and the frequency of such biopsies should be reevaluated in the light of their low yield with current immunosuppression. PMID- 18408577 TI - Recovery of heart function in children with acute severe heart failure. AB - BACKGROUND: The prognosis of acute heart failure is such that many children are considered for transplantation. Recovery of severe heart failure in a proportion of patients diagnosed with either dilated cardiomyopathy or myocarditis is well recognized, and this complicates the assessment for transplantation. There is little data on the time scale of recovery of heart function in children. OBJECTIVES: To describe the time course over which echocardiographic improvement of systolic function occurred in a cohort of children who presented in acute heart failure, without structural or metabolic abnormality. METHODS: Children with a first presentation of acute severe heart failure between 1990 and 2005. Time from presentation to the echocardiogram before left ventricular fractional shortening (FS) improved to 20% and 30% (complete recovery) was recorded. RESULTS: Twenty-seven children (11 male) were identified, and all had an initial FS <15%. Twenty-one patients required intravenous inotropes and three patients required extracorporeal membrane oxygenation. Seven patients had been on the active transplant list for a mean duration of 155 days. Four patients had probable viral myocarditis. Mean age at presentation was 15.7 (range, 0.1-72) months. Mean time to an FS of 20% was 3.6 (0.2-18) months and to 30% was 8.9 (0.7 24) months. Complete recovery occurred within 6, 9, 12, 18, and 24 months of presentation in 44%, 55%, 66%, and 96%, respectively. There was no correlation between age of presentation and length of time to recovery. CONCLUSIONS: Complete recovery of left ventricular systolic function is often delayed to more than 1 year from presentation. This may have major implications for timing of transplantation in an era where prolonged mechanical cardiac support is feasible even in infants. PMID- 18408578 TI - The use of donor and recipient screening for toxoplasma in the era of universal trimethoprim sulfamethoxazole prophylaxis. AB - BACKGROUND: Toxoplasmosis is a serious complication of solid organ transplantation. The highest risk of infection and disease occurs in heart recipients with primary infection transmitted by a seropositive donor to a seronegative recipient (donor-recipient mismatch). Toxoplasmosis has been reported to occur in noncardiac transplant recipients; however, no large studies examining the frequency of such events or the need for serologic screening exist. METHODS: A retrospective cohort study of 1,006 solid organ transplant recipients transplanted in our center between 1984 and 1997 was performed to examine the incidence of Toxoplasma seroconversion, reactivation, and clinical toxoplasmosis and to evaluate the impact of trimethoprim sulfamethoxazole (TMP/SMX) prophylaxis on these outcomes. RESULTS: Pretransplant Toxoplasma seroprevalence was 13.4% in donors and 17.8% in recipients. The incidence of Toxoplasma donor-recipient mismatch was 9.5% during the 14-year study period, and only 39.1% of mismatched recipients received TMP/SMX prophylaxis. Only four patients seroconverted, of whom two had received prophylaxis. There were no cases of clinical disease; either primary or reactivation. CONCLUSIONS: We therefore conclude that in transplant centers with low Toxoplasma seroprevalence, routine screening for Toxoplasma in solid organ donors and recipients is not necessary, particularly in the era of routine TMP/SMX prophylaxis. PMID- 18408579 TI - Assessing the value of ureteral stent placement in pediatric kidney transplant recipients. AB - BACKGROUND: Ureteral stent placement at kidney transplantation may reduce stenosis or leakage (S/L) complication rates. However, stent placement may also increase risk for early urinary tract infection (early UTI; <3 months after transplant) and BK virus allograft nephropathy (BKVAN). In children, the usefulness of stent placement is not well defined. METHODS: We analyzed retrospective data from children transplanted at our center for the three above outcomes in relation to stents. At our center, stent placement decision is driven by surgeon preference. RESULTS: Among 129 transplants from 1996 to 2006, early UTI was seen in 9.3% and S/L in 4.6%. By univariate analyses, stent placement was a significant risk factor for early UTI (P=0.0399) but not protective for S/L (P=0.23). In multivariate analyses, stent placement, human leukocyte antigen match, and bladder augmentation increased the odds ratio for early UTI. Only deceased donor source increased the odds ratio for S/L. In a truncated data set from 1999 to 2006, BKVAN occurred in 9 of 93 (9.6%). Per minute increase in warm ischemia time was the only significant risk factor for BKVAN by both univariate and Cox regression analyses. Stent placement did not improve graft survival (P=0.5726) but required general anesthesia for removal in the operating room, leading to additional cost and potential risk. CONCLUSION: Routine stent placement in children in this era of low urological complication rates and BKVAN needs reevaluation. PMID- 18408580 TI - Benefit of Neoral C2 monitoring in de novo cardiac transplant recipients receiving basiliximab induction. AB - BACKGROUND: For cyclosporine (CsA), 2-hr postdose level (C2) is the best single time point predictor of the area under the curve and a critical measure for effective dosing. The therapeutic CsA microemulsion (Neoral) C2 range in de novo heart transplant patients remains to be determined. PURPOSE: The purpose of this study was to determine the efficacy of CsA C2 monitoring in de novo heart transplant patients receiving basiliximab induction. METHODS: This prospective, multicenter, randomized study enrolled 87 adult heart transplant recipients stratified according to 4 to 6 hrs posttransplant serum creatinine less than or equal to 170 micromol/L (cohort A) or more than 170 micromol/L (cohort B). Patients in cohort A were randomized into three C2 ranges (A1: "high" n=25, 1600 1800 ng/mL; A2: "intermediate" n=27, 1400-1600 ng/mL; and A3: "low" n=24, 1200 1400 ng/mL). Patients in cohort B were randomized into intermediate (n=5) and low C2 (n=6). Target ranges were progressively lowered after 1 month. Immunosuppression included basiliximab, Neoral, mycophenolate mofetil, and corticosteroids. Endpoints were acute rejection and renal function. RESULTS: The incidence of acute rejection at 12 months was 44% in group A1, 41% in group A2, 33% in group A3, and 27% in cohort B. Pretransplant and 12-month creatinine clearance (mL/min) were group A1, 72+/-25 and 64+/-24; group A2, 81+/-32 and 68+/ 25; group A3, 91+/-28 and 86+/-26; and cohort B, 62+/-28 and 79+/-37. CONCLUSION: These results suggest that C2 monitoring is safe in de novo heart transplant patients. A low Neoral C2 range in combination with basiliximab induction resulted in preserved renal function without increased risk of acute rejection. PMID- 18408581 TI - Case-control comparison of at-home and hospital care for allogeneic hematopoietic stem-cell transplantation: the role of oral nutrition. AB - BACKGROUND: Acute graft-versus-host disease (GVHD) was reduced using home care compared with hospital care after allogeneic hematopoietic stem-cell transplantation (ASCT). METHODS: Between March 1998 and December 2006, 601 patients underwent ASCT at our unit. Requirements for at-home ASCT were fulfilled by 76 patients. A control group of 76 patients treated in the hospital were matched for age, sex, diagnosis, stage of disease, conditioning, stem-cell source, type of donor, and immunosuppression. Oral nutrition was determined as median kcal/kg/day for the first 21 days after ASCT. RESULTS: The home-care patients received more oral nutrition per day than hospital controls (P<0.05). Number of days at home correlated with oral nutrition (P=0.004). In multivariate analysis, acute GVHD of grades II to IV was associated with poor oral nutrition (P=0.003) and hospital care (P=0.06). Transplant-related mortality was associated with acute GVHD grades II to IV (P<0.0001) and bacteremia (P=0.004). In addition to acute GVHD and bacteremia, death was associated with absence of chronic GVHD (P=0.012). Five-year survival was 65% in patients treated at home, when compared with 47% in the controls (P=0.04). CONCLUSION: Better oral nutrition may be one reason for the reduced probability of acute GVHD and better survival with at-home care than with hospital care. PMID- 18408582 TI - BK virus nephropathy in renal transplant patients in London. AB - BACKGROUND: BK nephropathy (BKN) is an important cause of renal transplant dysfunction, believed to be associated with higher levels of immunosuppression. We assessed the experience of BKN in renal transplant patients in the London region. METHODS: All six London transplant centers participated and case notes of patients with BKN in 2004 to 2005 were reviewed. RESULTS: There were 17 cases of BKN, giving an incidence of 2.1%. Median time to diagnosis was 9 months. Median baseline creatinine rose from 150 to 196 mumol/L. At diagnosis, 16 patients were on tacrolimus, 15 on mycophenolate mofetil, and 10 on triple therapy with tacrolimus, mycophenolate mofetil, and prednisolone. Management of BKN involved reducing immunosuppression; cidofovir was used in two patients and methylprednisolone in five for acute rejection. Median follow-up time was 29.2 months. Creatinine returned to baseline in four patients, remained elevated in 12 and one patient lost his graft. The new median baseline creatinine was 216 mumol/L. Eight patients underwent repeat biopsies of which four became negative for BKV and three subsequently cleared the virus on blood and urine polymerase chain reaction and urine decoy cells. Overall, eight patients cleared the virus. None of age, sex, viral load, or biopsy characteristics (Banff ct score, Drachenberg grade, and number of BKV positive cells) were associated with poorer outcome when patients with increase in creatinine of less than 30% (n=7) or more than 30% (n=10) from baseline were compared. CONCLUSION: The incidence of BKN in this study is comparable with previous studies, with more favorable outcomes. It supports the association of BKN with potent immunosuppression. PMID- 18408583 TI - Evidence for the pivotal role of endogenous toll-like receptor 4 ligands in liver ischemia and reperfusion injury. AB - BACKGROUND: Although toll-like receptor 4 (TLR4) activation has been demonstrated to play a key role in the induction of intrahepatic inflammation, leading to hepatocellular damage in liver ischemia/reperfusion injury (IRI), the nature of TLR4 ligands generated during tissue injury remains to be elucidated. We hypothesized that endogenous TLR4 ligands, rather than endotoxin (lipopolysaccharide [LPS]), are instrumental in the activation of liver TLR4 leading to local inflammation response that culminates in ultimate organ IRI. METHODS AND RESULTS: By using the LPS-neutralizing agent, recombinant bactericidal/permeability-increasing protein, we showed that the endotoxin blockade failed to protect mouse livers from warm IRI, as assessed by serum alanine aminotransferase levels, intrahepatic inflammatory gene induction profile, and liver pathology. The recombinant bactericidal/permeability increasing protein did not cause any hepatocytoxicity by itself if injected into normal naive mice. Furthermore, we demonstrated that liver perfusates, generated by isolated liver perfusion system, contained LPS-independent, heat-sensitive protein molecules that activated macrophages to produce tumor necrosis factor (TNF)-alpha through TLR4 but not TLR2 pathway. CONCLUSION: This study provides a definitive evidence that endogenous TLR4 ligands are critical in the pathogenesis of liver IRI. PMID- 18408584 TI - Intensified postgrafting immunosuppression failed to assure long-term engraftment of dog leukocyte antigen-identical canine marrow grafts after 1 gray total body irradiation. AB - BACKGROUND: Late graft rejection after conditioning with 1 Gy of total body irradiation (TBI) was consistently seen in historical dogs given two postgrafting immunosuppressive drugs. METHODS: Here, 16 dogs were given four different three drug combinations of cyclosporine, mycophenolate mofetil, sirolimus, or methotrexate after 1 Gy TBI and dog leukocyte antigen-identical marrow grafts. In addition, we assessed the effects of TBI doses of 0.5, 1.0, 2.0, or 3.0 Gy, respectively, on immune functions in six dogs not given marrow grafts. RESULTS: All dogs showed initial engraftment, 13 rejected, and three had sustained grafts beyond 26 weeks. The dogs with durable grafts had received greater median numbers of nucleated marrow cells compared with the 13 dogs that rejected their grafts (6.14 vs. 3.6 x 10(8) per kg; P=0.03). In a Cox proportional hazard model, which included data from 16 historical dogs, each increase in transplanted marrow cell numbers by 1 x 10(8) per kg decreased the hazard ratio of rejection by 0.5. Decreasing percents of remaining CD3, CD4, and CD8 cells in peripheral blood and lymph nodes were observed with increasing TBI doses. Further, greater suppressions of B-cell- and T-cell-dependent production of IgM and IgG antibodies in response to sheep red blood cell injections were observed after 2 Gy compared with 1 Gy TBI. CONCLUSION: Overall, triple postgrafting immunosuppression after 1 Gy TBI was well tolerated but failed to prevent graft rejection in this model. In vivo radiation studies have shown higher numbers of remaining host lymphocytes and better T-cell-dependent antibody production after 1 Gy compared with 2 Gy TBI. PMID- 18408585 TI - Phenotypic and functional differences between wild-type and CCR2-/- dendritic cells: implications for islet transplantation. AB - BACKGROUND: Trafficking of dendritic cells (DC), the primary regulators of alloimmune responses, is controlled by chemokines. Here, we provide evidence that lack of CCR2 could lead to the generation of functionally and phenotypically different DC, which in part could explain the benefits observed in transplanting islets in CCR2 recipients. METHODS AND RESULTS: We show that, in contrast to the in vitro DC maturation model, in vivo DC maturation is accompanied by an increase in the expression of CCR2. Compared with wild-type (WT), DC generated in vitro from CCR2 mice, and DC extracted from CCR2 naive mice or from CCR2 recipients of islet allografts, display lesser allostimulatory capacity. Compared with WT DC, CCR2 DC produce more IL-4 and induce more IL-4-producing T cells. CCR2 DC also promote the generation of regulatory T cells that more efficiently suppress T cell proliferative responses by mixed leukocyte reaction. Similarly, the percentage of CD4CD25FoxP3 cells were found to be higher in CCR2 recipients of islet allografts than in WT recipients. CONCLUSIONS: In summary, lack of CCR2 interferes with the allostimulatory capacity of DC and promotes the generation of regulatory T cells. This is the first demonstration of a mechanistic link between targeting a specific chemokine pathway and the DC-regulatory T cell axis in alloimmunity. PMID- 18408586 TI - TNF receptor independent activation of the cytomegalovirus major immediate early enhancer in response to transplantation. AB - BACKGROUND: Reactivation of latent human cytomegalovirus (HCMV) infection is a significant risk factor for long term allograft dysfunction. The molecular pathways involved in reactivation of latent virus have not been identified. Previous studies suggested that tumor necrosis factor (TNF) -mediated activation of nuclear factor of kappa light polypeptide gene enhancer in B-cells (NF-kappa B) leading to transcriptional reactivation of viral immediate early (ie) gene expression might be important in transplant-associated viral reactivation. CMV IE gene expression is controlled by the major immediate early promoter/enhancer (MIEP). Because HCMV does not infect mice, transgenic mice carrying a beta galactosidase reporter gene under the control of the HCMV immediate early enhancer (MIEP-lacZ mice) are a valuable model for studying regulation of CMV IE gene expression in vivo. We have used TNF receptor-deficient MIEP-lacZ (MIEP-lacZ TNFR DKO) mice to study the requirement for TNF in transplant-induced activation of the MIEP. METHODS: Allogenic kidney transplants were performed using MIEP-lacZ TNFR DKO or MIEP-lacZ TNFRwild-type donor mice. beta-Galactosidase activity was used to measure activation of the IE enhancer in donor kidneys at 2 days of posttransplantation and in contralateral controls. Transcription factor activation was assayed with Trans-Am kits. RESULTS: Allogenic and syngenic transplantation activate the HCMV IE enhancer to the same extent. TNF receptor signaling was not required for activation of the MIEP. TNF receptor signaling was required for activation of NF-kappaB, but not for activation of activating protein 1 family members junD and Fra-1 in day 2 allografts. CONCLUSIONS: TNF independent pathways can activate the enhancer in response to allogenic transplantation. This may occur through activation of MIEP-binding transcription factors other than NF-kappa B. PMID- 18408587 TI - Clinical importance of anti-human leukocyte antigen-specific antibody concentration in performing calculated panel reactive antibody and virtual crossmatches. AB - BACKGROUND: Highly sensitized patients develop multi-human leukocyte antigen (HLA) specific antibodies. This study measures concentrations of anti-HLA antibodies in multispecific sera by converting fluorescence intensity into molecules of equivalent soluble fluorochrome (MESF) units. This was used to determine MESF units required for a positive T and B flow crossmatches (FLXM). METHODS: MESF values of negative controls and sera from patients devoid of HLA antibodies were measured by FLXM and flow panel reactive antibody (PRA) screening beads. Fluorescence intensity values of anti-HLA specific antibodies determined by FlowPRA single antigen beads of highly sensitized patients were converted into MESF units. In addition, endpoint titers, MESF units, and % PRA of 26 sera were established. RESULTS: MESF analysis accurately predicted the outcomes of 100% of T and B FLXM of sera with strong (MESF units>18,000) donor-specific antibody (DSA). The predictive values of T and B FLXM declined to 95% and 88% with weak DSA (6,000 MESF<10,000). Endpoint titers of sera from highly sensitized patients ranged from 1:512 to 1:8 with corresponding MESF values of 452,596 to 20,000 units. However, there was no statistical difference in PRA values among these sera (95%-100%). We successfully transplanted five patients who had weak donor specific HLA antibodies (MESF units>2,000). The graft survival at 1 year was 100% and there was no evidence of DSA posttransplant. CONCLUSION: MESF analysis is both a time and cost efficient way of measuring antibody strength. The strength of the antibody present in the sera of transplant candidates is critical for crossmatch prediction. PMID- 18408588 TI - A blocking anti-CD28-specific antibody induces long-term heart allograft survival by suppression of the PKC theta-JNK signal pathway. AB - This study investigated the effects of a blocking anti-CD28 antibody (Anti-CD28 PV1-IgG3) in vitro and in vivo. Anti-CD28-PV1-IgG3, a hamster-mouse chimeric antibody against murine CD28, which does not provide CD28-positive signaling during TCR-driven T cell activation, enabled long-term survival of heart allografts across a complete mismatch of the MHC in rats. Among the T cell signaling proteins tested in the spleens from recipients, we found that recipients treated with anti-CD28-PV1-IgG3 exhibited suppression of alloantigen initiated proximal TCR signaling events, including Lck, Zap70, Vav, and PI3K expression, and their PKC theta- and JNK-regulated expression/activation. This leads to attenuation of intragraft T cell infiltration and expression of T cell effector molecules. These results indicate that targeting the CD28 receptor with a blocking antibody leads to long-term allograft survival by reducing activation of alloantigen-mediated key signaling events in T cells that might be crucial for full T cell activation. PMID- 18408589 TI - Bronchoalveolar immunologic profile of acute human lung transplant allograft rejection. AB - Bronchoalveolar lavage fluid (BALF) offers a potential means to diagnose acute rejection and could provide insight into the immune mechanisms responsible for lung allograft rejection. Transbronchial biopsies from 29 bronchoscopic procedures were assessed for rejection. Concurrent BALF lymphocyte subsets were examined by flow cytometry, including CD4 and CD8 T cells and their activation status by CD38 expression, natural killer (NK), NK-like T (NT), B, regulatory T, and invariant receptor NK-T cells. Percentages of CD4 were reduced, and CD8 and activation of CD4 T cells correlated with rejection. There were trends for increased NT, reduced NK, and increased B cell percentages with rejection, suggesting potential roles of these cells. Among regulatory cells, the percentages of regulatory T cells decreased and CD4/CD8 invariant NK-T cells increased during rejection, suggesting a proinflammatory profile. A unique BALF lymphocyte profile was associated with rejection and may provide insight into the pathogenesis of allograft rejection. PMID- 18408590 TI - Role of invariant natural killer T cells in liver sinusoidal endothelial cell induced immunosuppression among T cells with indirect allospecificity. AB - We have previously demonstrated that liver sinusoidal endothelial cells (LSECs) that endocytose portally injected allogeneic cells exert immunosuppressive effects on T cells with indirect allospecificity. In this study, we have demonstrated that invariant natural killer T cells plays a significant role in such immunosuppressive effects induced by LSECs. The endocytic activity of LSECs toward intraportally injected splenocytes from B6 major histocompatibility complex class II-deficient C2ta (C2D) mice was markedly impaired in BALB/c CD1d deficient mice. The intraportal adoptive transfer of LSECs isolated from BALB/c wild-type mice treated with a portal injection of B6 C2D splenocytes into BALB/c mice significantly prolonged the survival of subsequently transplanted heart allografts; however, the transfer of LSECs isolated from similarly treated BALB/c CD1d-deficient mice did not produce such a survival prolonging effect. These findings indicate that natural killer T cells are required for the LSEC-induced immune modulation of T cells with indirect allospecificity. PMID- 18408591 TI - Carcinoma "en Cuirasse" from cutaneous squamous cell carcinoma in a renal transplant patient. PMID- 18408592 TI - Mineralocorticoid receptor and 11beta-hydroxysteroid dehydrogenase type II expression in renal cell neoplasms: a tissue microarray and quantitative RT-PCR study. AB - The kidney is an important target for mineralocorticoids. Aldosterone, the major endogenously secreted mineralocorticoid, acts by binding to mineralocorticoid receptor (MR) in the distal renal tubule. The enzyme 11beta-hydroxysteroid dehydrogenase type II (11beta-HSD2) prevents the binding of glucocorticoids to the MR by inactivating cortisol to cortisone. Our goal was to determine whether MR and 11beta-HSD2 expression could be used to characterize the major types of renal cell neoplasms. Using immunohistochemistry we analyzed tissue microarray specimens from 132 patients with renal cell neoplasms, stratified into 84 clear cell renal cell carcinomas (CRCC), including 9 cases clear cell carcinoma with predominantly granular cytoplasm; 14 papillary RCC (PRCC); 20 chromophobe RCC (CHRCC); and 14 oncocytomas (OCs). MR and 11beta-HSD2 expression were also quantitated by real-time reverse transcription-polymerase chain reaction. Expression of both MR and 11-betaHSD2 was detected in the distal nephrons of normal kidneys. The CHRCC group stained for 11-betaHSD2 in a membranous and cytoplasmic pattern whereas diffuse cytoplasmic reactivity was seen in OCs. MR and 11beta-HSD2 were coexpressed in most of CHRCC (90% and 95%) and oncocytomas (93% and 100%). No MR staining was detected in CRCC, including clear cell carcinoma with predominantly granular cytoplasm, or in PRCC. Only 2 cases of CRCC (2.6%) showed focal positivity for 11beta-HSD2, whereas all PRCCs were negative. CHRCC and OC demonstrated significantly higher levels of MR and 11beta-HSD2 expression than CRCC and PRCC by real-time polymerase chain reaction. Moreover, CHRCC showed higher expression of MR and 11beta-HSD2, as compared with OC. Our study indicates MR and 11beta-HSD2 are both sensitive and specific markers of the distal nephron and its related neoplasms (CHRCC and OC). Additionally, the staining pattern and the level of MR and 11beta-HSD2 expression seems to be useful in the distinction of CHRCC from OC. MR and 11beta-HSD2 should be considered in the immunohistochemical panel to more accurately subtype renal cell tumors. PMID- 18408593 TI - Microscopic gastrointestinal stromal tumors in esophageal and intestinal surgical resection specimens: a clinicopathologic, immunohistochemical, and molecular study of 19 lesions. AB - Microscopic gastrointestinal stromal tumors (GISTs) (synonyms: sporadic interstitial cell of Cajal hyperplasia, seedling GISTs, minimal GISTs) are common incidental findings in gastroesophageal resections (9% to 35%). To our knowledge, their frequency, clinicopathologic features, and molecular pathogenesis from nongastroesophageal sites have so far not been sufficiently analyzed. We studied 19 lesions from distal esophagus (n=8), gastroesophageal junction (n=2), sigmoid colon (n=5), and vermiform appendix, cecum, rectum, and small intestine (1 each). Microscopic GISTs were detected in 0.2%, 0.1%, and 0.01% of routinely processed resection specimens from sigmoid colon, vermiform appendix, and rectum, respectively. Patients were 11 men and 8 women with a mean age of 66 years (range, 57 to 86 y). Thirteen patients had GI cancers and 5 had diverticular disease. None has a family history of GIST or features of neurofibromatosis 1. Lesions were 0.5 to 4 mm in size (mean, 1.12 mm), were all spindled and had noncircumscribed infiltrating borders. All arose in the muscularis propria and 2 were predominantly subserosal. Immunohistochemistry revealed a CD117/CD34/smooth muscle actin-negative phenotype in 18/19 lesions. Three KIT exon 11 mutations (2 point mutations and 1 deletion, all involving W557) were detected in 3/12 lesions with successful molecular analysis. In conclusion, incidental microscopic GISTs are uncommon in intestinal resections (< or =0.1%), contrasting with their gastroesophageal counterparts (> or =9%). Somatic KIT mutations are early initiating molecular events in a subset of them. The remarkable variation in the incidence of microscopic GISTs at different GI sites suggests an origin from heterogeneous subsets of interstitial cells of Cajal with varying potentials for neoplastic transformation. PMID- 18408594 TI - Gastrointestinal melanoma or clear cell sarcoma? Molecular evaluation of 7 cases previously diagnosed as malignant melanoma. AB - Clear cell sarcoma (CCS) is a rare tumor classically associated with the tendons and aponeuroses of distal extremities of young adults. CCS and malignant melanoma (MM) share immunohistochemical profiles and ultrastructural features, but classic CCS has characteristic morphology with low mitotic activity and minimal pleomorphism. Occasional cases show pleomorphism, high mitotic index, and/or melanin pigmentation, making CCS indistinguishable from MM based on morphology. However, CCS is genetically distinct owing to its consistent association with a t(12;22)(q13;q12) chromosomal translocation, leading to the formation of the EWS/ATF1 fusion transcript. This translocation has never been documented in cutaneous melanoma, and thus is regarded as specific for CCS. Recent evidence suggests that primary "malignant melanomas" in unusual anatomic sites, most notably the gastrointestinal (GI) tract, may be CCS. This is supported by 11 cases of primary GI CCS with the t(12;22) translocation. We used reverse transcription polymerase chain reaction and fluorescence in situ hybridization to examine whether a proportion of cases diagnosed as MM of the GI tract in patients without a history of cutaneous MM actually represent primary GI CCS. In total, we examined 7 cases: Four with no prior history of MM, 2 with histories of cutaneous MM, and 1 with an anal MM. All 4 cases for which there was no history of cutaneous/mucosal MM harbored the EWS/ATF1 fusion transcript. We report the largest series of GI CCS and have shown that molecular studies may be warranted in cases that otherwise seem to represent MM of unusual primary locations. PMID- 18408595 TI - Partial atrophy on prostate needle biopsy cores: a morphologic and immunohistochemical study. AB - Partial atrophy is the most common benign mimicker of prostate cancer on needle biopsy. Of 3916 prostate needle core biopsy cases received in our consultation service over a period of 3 months (March 1, 2007 to May 31, 2007), 170 cases (4.3%) with partial atrophy were diagnosed as atypical glands by outside pathologists and prospectively identified. We supplemented our material with 108 cases of partial atrophy sent to our consultation service in 2006 from a single institution, which frequently uses a triple cocktail stain [p63, high molecular weight cytokeratin (HMWCK), alpha-methyl acyl-Coa racemase (AMACR)]. The morphologic features of the 278 cases and immunohistochemistry of 236 cases (198 with prostate cocktail and 38 with only basal cell makers) were analyzed. Forty eight of 278 (17.3%) partial atrophy cases were mixed with postatrophic hyperplasia. Enlarged nuclei were visible in 43/278 (15.5%) cases, with prominent nucleoli seen in 58/278 (20.9%) cases (30 cases associated with nuclear enlargement). Of 198 cases with a prostatic cocktail stain, 48 (24.2%) had a cancer pattern for both basal cells and AMACR (p63-, HMWCK-, and AMACR+), 14 (7.1%) had a cancer pattern for basal cells (p63-, HMWCK-, and AMACR-), 89 (44.9%) had a cancer pattern for AMACR (p63+, HMWCK+, and AMACR+), and 47 (23.7%) had a totally benign pattern (p63+, HMWCK+, and AMACR-). Of the 198 cases using the cocktail stain, 136 (68.7%) had positive basal cell staining. The percentage of basal cells labeled with the combination of p63/HMWCK was: <5% in 42 (21.2%) cases, 5% to 75% in 58 (29.3%) cases, and >75% in 36 (18.2%) cases. An additional 38 cases immunostained only for p63 and/or HMWCK was negative in 2 (5.2%) cases, <5% (13.1%) in 5 cases, 5% to 75% in 19 (50%) cases, and >75% in 12 (31.6%) cases. In conclusion, partial atrophy is a benign mimicker of adenocarcinoma both as a result of its routine morphologic features and its immunohistochemical profile. Recognition of the classic morphology of partial atrophy on routine hematoxylin and eosin-stained sections is critical to avoid misdiagnosing partial atrophy as adenocarcinoma. PMID- 18408596 TI - Alterations in the immunohistochemical expression of Das-1 and CG-3 in colonic mucosal biopsy specimens helps distinguish ulcerative colitis from Crohn disease and from other forms of colitis. AB - Distinction between ulcerative colitis (UC) and Crohn disease (CD) in mucosal biopsies is often difficult. Das-1 and CG-3 are monoclonal antibodies directed against an unknown colonic epithelial protein and human tropomyosin isoform-5, respectively, both show altered expression in patients with UC. In this study, we evaluated the utility of Das-1 and CG-3 in distinguishing UC from CD and from other types of colitis. One colonic biopsy specimen from each of 85 patients with confirmed UC (n=25), CD (n=15), lymphocytic (n=15), collagenous (n=15), and ischemic (n=15) colitis, and also 10 samples from normal controls, were stained for Das-1 and CG-3 using standard techniques. Reactivity for Das-1 and CG-3 was noted to be absent or present, and the location (ie, surface+/-crypt epithelium) and degree (weak or strong) of CG-3 staining was recorded. Loss of Das-1 staining occurred more frequently in UC (96%) compared with CD (20%), lymphocytic (20%), collagenous (13%), and ischemic colitis (0%) cases, as well as controls (10%, P<0.001 for all comparisons). CG-3 positivity in crypt epithelium was significantly more common in UC (52%) compared with the other groups (P< or =0.02 for all comparisons). The combination of strong crypt CG-3 staining and loss of Das-1 staining was noted in 44% of UC cases, but not in any other type of colitis (P=0.003 for all comparisons). We conclude that the patterns of Das-1 and CG-3 staining in colonic mucosal biopsies may be clinically useful in distinguishing UC from CD and from other colitidies. PMID- 18408598 TI - Prediction of activity mode with global positioning system and accelerometer data. AB - PURPOSE: The primary aim of this pilot study was to assess how well the combination of global positioning system (GPS) and accelerometer data predicted different activity modes. METHODS: Ten adults (seven male, three female; 23-51 yr) simultaneously wore a GPS unit and accelerometer during bouts of walking, jogging/running, bicycling, inline skating, or driving an automobile. Discriminant function analysis was used to identify a parsimonious combination of variables derived from accelerometer counts and steps and GPS speed that best classified mode. A total of 29 bouts were used to develop this classification criterion. This criterion was validated using two datasets generated from the complete collection of minute-by-minute values from all bouts. RESULTS: Model development with "calibration" data showed that two accelerometer variables alone (median counts and steps) resulted in 26 of 29 bouts (90%) being correctly classified. Prediction of activity mode using counts and steps in a minute-by minute "validation" dataset (N = 200) was 86.5%. Using three variables from the accelerometer and GPS (median counts, steps and speed) resulted in correct classification in 27 of 29 activity bouts in the "calibration" data (93%). In the "validation" dataset comprising 200 min, the combination of accelerometer counts and steps and GPS speed were able to correctly classify 91% of the observations. Walking and bicycling minutes were correctly classified most frequently (96%). In another "validation" dataset consisting of activity bouts, this combination of variables resulted in correct classification in 42 of 43 bouts (98%). CONCLUSION: This pilot study provides evidence that the addition of GPS to accelerometer monitoring improves physical activity mode classification to a small degree. Larger studies among free-living individuals and with an expanded range of activities are needed to replicate the current findings and further determine the merits of using GPS with accelerometers for mode identification. PMID- 18408599 TI - Contextual barriers to lifestyle physical activity interventions in Hong Kong. AB - PURPOSE: Increased lifestyle physical activity, for instance, use of active transport, is a current public health target. Active transport interventions that target stair climbing are consistently successful in English-speaking populations yet unsuccessful in Hong Kong. We report two further studies on active transport in the Hong Kong Chinese. METHODS: Pedestrians on a mass transit escalator system (study 1) and in an air-conditioned shopping mall (study 2) were encouraged to take the stairs for their cardiovascular health by point-of-choice prompts. Observers coded sex, age, and walking on the mass transit system, with the additional variables of presence of children and bags coded in the shopping mall. In the first study, a 1-wk baseline was followed by 4 wk of intervention (N = 76,710) whereas in the second study (shopping mall) a 2-wk baseline was followed by a 2-wk intervention period (N = 18,257). RESULTS: A small but significant increase in stair climbing (+0.29%) on the mass transit system contrasted with no significant changes in the shopping mall (+0.09%). The active transport of walking on the mass transit system was reduced at higher rates of humidity and temperature, with steeper slopes for the effects of climate variables in men than in women. CONCLUSIONS: These studies confirm that lifestyle physical activity interventions do not have universal application. The context in which the behavior occurs (e.g., climate) may act as a barrier to active transport. PMID- 18408600 TI - Maximal physiological responses between aquatic and land exercise in overweight women. AB - PURPOSE: To investigate the maximal physiological responses between aquatic and land-based graded exercise tests in overweight women. METHODS: Twenty healthy, overweight (body mass index (BMI) > or = 25 kg.m(-2)), Caucasian women (mean +/- SD; age 48 +/- 7 yr, BMI 30 +/- 4 kg.m(-2)) completed a deep water running (DWR) and treadmill walking (TMW) graded exercise test. Maximal responses during the DWR and TMW graded exercise tests were compared using paired t-tests. Comparisons were made in the incidence of achievement of maximal oxygen consumption (VO2max) criteria between DWR and TMW protocols. Criteria were a plateau in VO2 (change < 2.1 mL.kg.min(-1)), heart rate (HR) equal to or above the age-adjusted maximum, and respiratory exchange ratio (RER) > or = 1.15. RESULTS: Maximal responses for VO2max (22.5 +/- 4.86 vs 27.7 +/- 4.73 mL.kg.min(-1)), HRmax (159 +/- 16 vs 170 +/- 12 bpm), and RER (1.03 +/- 0.06 vs 1.10 +/- 0.06) were significantly lower (P < 0.01) for the DWR test compared with the TMW test, respectively. Achievement of various VO2max criteria was demonstrated more consistently during the TMW test than the DWR test. CONCLUSION: Maximal physiological responses of overweight women to DWR and TMW are significantly different but are comparable with other populations. As the maximal responses for DWR compared with TMW differ, the use of land-based criteria for VO2max is not recommended for a graded DWR exercise test. PMID- 18408601 TI - Body composition and fitness during strength and/or endurance training in older men. AB - PURPOSE: This study examined adaptations in body composition and physical fitness during a 21-wk strength and/or endurance training period in 40- to 65-yr-old men. We also compared the usefulness of different methods for the analysis of body composition to detect training-induced adaptations. METHODS: Fifty-three men were randomized into the endurance training (E: N = 14), strength training (S: N = 13), combined strength and endurance training (SE: N = 15), or control (C: N = 11) groups. S and E trained 2 and SE 2 x 2 times a week for strength and endurance. RESULTS: Percentage of fat (fat%) decreased (5-8%) similarly in all training groups. Fat% measured by DXA at baseline and its change correlated with those recorded by bioimpedance (r = 0.90 and 0.66), skinfolds (r = 0.80 and 0.78), and waistline (r = 0.84 and 0.74). Lean mass in legs (DXA) increased only in S (2.0 +/- 1.5%, P < 0.001), but the thickness of vastus lateralis and intermedius measured by ultrasound increased (7-11%) in all training groups, and that of triceps brachii increased in S (22%) and SE (20%). Maximal concentric force increased significantly in S, SE, and E (by 22, 23, and 7%), and maximal oxygen uptake increased in both E (11%) and SE (11%). CONCLUSIONS: Waist circumference and skinfold thickness seem to reasonably assess changes in percent body fat during training. However, only DXA was capable to separate small differences between the groups in training-induced changes in lean body mass. Combined strength and endurance training is of greater value than either alone in optimizing body composition or improving physical fitness in older men. PMID- 18408602 TI - Evaluation of ultrasound in assessing body composition of high school wrestlers. AB - PURPOSE: To evaluate the accuracy of ultrasound (ULTRA) in assessing fat-free mass (FFM) in comparison with hydrostatic weighing (HW) and skinfolds (SK) in high school wrestlers in a hydrated state. METHODS: Body composition was determined by ULTRA, HW, and three-site SK in 70 high school wrestlers (mean +/- SD: age, 15.5 +/- 1.5; height, 1.60 +/- 0.08 m; body mass, 65.8 +/- 12.7 kg). For all methods, body density (Db) was converted to percent body fat (%BF) using the Brozek equation. Hydration state was quantified by evaluating urine specific gravity. RESULTS: There were no significant differences for estimated FFM between ULTRA (57.2 +/- 9.7 kg) and HW (57.0 +/- 9.9 kg); however, SK (54.9 +/- 8.8 kg) were significantly different from HW. The standard errors of estimate for FFM with HW as the reference method were 2.40 kg for ULTRA and 2.74 kg for SK. Significant correlations were found for FFM between HW and ULTRA (r = 0.97, P < 0.001) and between HW and SK (r = 0.96, P < 0.001). A systematic bias was found for SK, as the difference between SK and HW significantly correlated with the FFM average of the two methods (r = -0.38, P < 0.001). This systematic bias was not found for ULTRA (r = - 0.07). CONCLUSIONS: This study demonstrates that ULTRA provides similar estimates of FFM when compared with HW in a heterogeneous high school wrestling population during a hydrated state. ULTRA should be considered as an alternative field-based method of estimating the FFM of high school wrestlers. PMID- 18408603 TI - Effect of match-related fatigue on short-passing ability in young soccer players. AB - PURPOSE: To examine whether the fatigue accumulated during match play or determined by short bouts of high-intensity intermittent activities affect short passing ability in junior soccer players. A further aim was to examine the influence of physical fitness as measured using the Yo-Yo Intermittent Recovery Test (YYIRT) on the changes in short-passing ability after a 5-min simulation of high-intensity activities (HIS). METHODS: Sixteen players (mean +/- SD: age 17.6 +/- 0.5 yr, height 174 +/- 7 cm, body mass 68 +/- 6 kg) participated in the study. A quasi-experimental control-period design was used for the study. Short passing ability was measured using the Loughborough Soccer Passing Test (LSPT). Players completed the LSPT in two sessions during the 1-wk control period, followed by two unofficial matches during which the LSPT was performed during and after the first and the second halves of the game. Furthermore, the change in LSPT performance was determined after 5 min of HIS. RESULTS: A decline in LSPT performance was found during and after the game (P < 0.01). The accuracy of the LSPT decreased after the HIS. A significant correlation was found between the YYIRT scores and the decline in LSPT performance (accuracy, total time, total time with penalties) after HIS (r = -0.51 to -0.65; P < 0.05). CONCLUSIONS: This study showed that the fatigue developed during a match and after relatively short bouts of high-intensity intermittent activities has a detrimental effect on short passing ability, and that the fatigue-related decline in technical proficiency for a given intensity is associated with the fitness level of the players. PMID- 18408604 TI - Effect of lengthening contraction velocity on muscle damage of the elbow flexors. AB - PURPOSE: This study investigated the effect of lengthening contraction velocity on exercise-induced muscle damage. METHODS: Sixteen men were placed into two groups performing either 30 (N = 8) or 210 (N = 8) maximal lengthening contractions of the elbow flexors on an isokinetic dynamometer. Dominant and nondominant arms were randomly assigned for a slow-velocity (S: 30 degrees.s(-1)) or a fast-velocity (F: 210 degrees.s(-1)) exercise separated by 14 d. Maximal voluntary strength of isometric contractions (iMVC) and isokinetic concentric contractions (cMVC), range of motion (ROM), upper-arm circumference, muscle soreness, and serum creatine kinase (CK) activity were measured before, immediately after, and 1-120 h after exercise. Changes in these measures over time were compared by a two-way repeated-measures ANOVA to examine the effect of velocity in the same number of contractions (S30 vs F30; S210 vs F210) or the effect of contraction number at the same velocity (S30 vs S210; F30 vs F210). RESULTS: A significant (P < 0.05) interaction effect was evident only for iMVC between S30 and F30, but it was evident for iMVC, cMVC, ROM, and CK between S210 and F210. Changes in most of the measures were significantly (P < 0.05) smaller after 30 contractions (S30 and F30) than after 210 contractions (S210 and F210). CONCLUSION: These results suggest that the effect of contraction velocity on the magnitude of muscle damage after 30 contractions is minor; however, when 210 lengthening contractions were performed, the effect of contraction velocity became conspicuous. It is concluded that fast-velocity lengthening contractions are likely to induce greater muscle damage than slow-velocity contractions; however, muscle fatigue seems to be a confounding factor for the velocity effect. PMID- 18408605 TI - Effect of maturation on the relationship between muscle size and force production. AB - PURPOSE: Although it is well accepted that an increase in muscle size is linked to an increase in muscle force, the relationship between muscle size and maximal strength during maturation is still discussed. In the present study we aimed at determining whether maturation affects the relationship between muscle size and maximal strength, and we investigated the reasons accounting for the discrepancies among previous studies. METHODS: Maximal isometric handgrip force (Fmax) and forearm muscle size were measured in 14 prepubertal boys (11.3 +/- 0.8 yr old), 16 adolescents (13.3 +/- 1.4 yr old), and 16 men (35.4 +/- 6.4 yr old). Anatomic maximal cross-sectional area (MCSA) and muscle volume (VM) were measured using MRI, and these results were compared with muscle volume (VL) obtained from anthropometric measurements. RESULTS: Fmax was linearly correlated with VM (r2 = 0.90), VL (r2 = 0.85), and MCSA (r2 = 0.87), while VM was strongly correlated with VL (r2 = 0.90). The Fmax/VM ratio did not differ among groups, whereas Fmax/VL and Fmax/MCSA ratios were significantly higher in adults than in children and adolescents. These results demonstrated that, when compared with MRI, anthropometric measurements led to a systematic overestimation of muscle volume. In addition, this overestimation was significantly larger in children (43.1%) and adolescents (38.5%) as compared with adults (20.5%) (P < 0.05). CONCLUSION: Our results showed that the maximal isometric strength exerted by the forearm muscles in humans is proportional to their size whatever the age, and that VM is the best index of muscle size during growth. The previously reported increased ability to produce maximal strength from childhood to adulthood could be explained by systematic bias introduced by the method used to characterize muscle size instead of physiological or neural changes. PMID- 18408606 TI - The influence of arch supports on knee torques relevant to knee osteoarthritis. AB - PURPOSE: Changes in footwear and foot orthotic devices have been shown to significantly alter knee joint torques thought to be relevant to the progression if not the development of knee osteoarthritis (OA) in the medial tibiofemoral compartment. The purpose of this study was to determine if commonly prescribed arch support cushions promote a medial force bias during gait similar to medial wedged orthotics, thereby increasing knee varus torque during both walking and running. METHODS: Twenty-two healthy, physically active young adults (age, 29.2 +/- 5.1 yr) were analyzed at their self-selected walking and running speeds in control shoes with and without arch support cushions. Three-dimensional motion capture data were collected in synchrony with ground reaction force (GRF) data collected from an instrumented treadmill. Peak external knee varus torque during walking and running were calculated through a full inverse dynamic model and compared. RESULTS: Peak knee varus torque was statistically significantly increased by 6% (0.01 +/- 0.02 N.m.(kg.m)(-1)) in late stance during walking and by 4% (0.03 +/- 0.03 N.m.(kg.m)(-1)) during running with the addition of arch support cushions. CONCLUSIONS: The addition of material under the medial aspect of the foot by way of a flexible arch support promotes a medial force bias during walking and running, significantly increasing knee varus torque. These findings suggest that discretion be employed with regard to the prescription of commonly available orthotic insoles like arch support cushions. PMID- 18408607 TI - Increased carbohydrate oxidation after ingesting carbohydrate with added protein. AB - PURPOSE: To examine the metabolic impact of including protein in a postexercise carbohydrate supplement when ingested between two bouts of prolonged running performed within the same day. METHODS: Six healthy men participated in two trials separated by 14 d, each involving a 90-min treadmill run at 70% VO2max followed by 4 h of recovery and a subsequent 60-min run at the same intensity. At 30-min intervals during recovery, participants ingested either a solution containing 0.8 g.kg(-1)h(-1) of carbohydrate (CHO) or the same solution plus an additional 0.3 g.kg(-1)h(-1) of whey protein isolate (CHO-PRO). Muscle biopsies were obtained from the vastus lateralis at the beginning and end of the recovery period, with a third muscle biopsy taken following the second treadmill run. RESULTS: Despite higher insulinemic responses to the CHO-PRO solution than to the CHO solution (P < 0.05), rates of muscle glycogen resynthesis during recovery were not different between treatments (CHO = 12.3 +/- 2.2 and CHO-PRO = 12.1 +/- 2.7 mmol glucosyl units per kilogram of dry mass per hour). Furthermore, there were no differences between treatments in muscle glycogen degradation during subsequent exercise (CHO = 2.2 +/- 0.3 and CHO-PRO = 2.0 +/- 0.1 mmol glucosyl units per kilogram of dry mass per minute). In contrast, whole-body carbohydrate oxidation during the second run was significantly greater with the CHO-PRO treatment than with the CHO treatment (48.4 +/- 2.2 and 41.7 +/- 2.6 mg.kg( 1)min(-1), respectively; P < 0.01). CONCLUSION: These data show that the inclusion of protein in a carbohydrate-recovery supplement can increase the oxidation of extramuscular carbohydrate sources during subsequent exercise without altering the rate of muscle glycogen degradation. PMID- 18408608 TI - Effect of dietary fat on serum and intramyocellular lipids and running performance. AB - PURPOSE: This study evaluated whether lowering IMCL stores via 3-d consumption of very-low-fat (LFAT) diet impairs endurance performance relative to a moderate-fat diet (MFAT), and whether such a diet unfavorably alters lipid profiles. METHODS: Twenty-one male and female endurance-trained runners followed a controlled diet and training regimen for 3 d prior to consuming either a LFAT (10% fat) or MFAT (35% fat) isoenergetic diet for another 3 d in random crossover fashion. On day 7, runners followed a glycogen normalization protocol (to equalize glycogen stores) and then underwent performance testing (90-min preload run at 62 +/- 1% VO2max followed by a 10-km time trial) on the morning of day 8. Muscle biopsies obtained from vastus lateralis before and after performance testing were analyzed for IMCL (via electron microscopy) and glycogen content (via enzymatic methodology). RESULTS: Despite approximately 30% lower IMCL (0.220 +/- 0.032% LFAT, 0.316 +/- 0.049% MFAT; P = 0.045) and approximately 22% higher muscle glycogen stores at the start of performance testing (P = 0.10), 10-km performance time was not significantly different following the two diet treatments (43.5 +/- 1.4 min LFAT vs 43.7 +/- 1.2 min MFAT). However, LFAT produced less favorable lipid profiles (P < 0.01) by increasing fasting triglycerides (baseline = 84.9 +/ 8.6; LFAT = 118.7 +/- 10.0 mg.dL(-1)) and the total cholesterol:HDL cholesterol ratio (baseline = 3.42 +/- 0.13:1; LFAT = 3.75 +/- 0.20:1), whereas MFAT lowered triglycerides (baseline = 97.5 +/- 12.2; MFAT = 70.9 +/- 7.1 mg.dL(-1)) and the total cholesterol:HDL cholesterol ratio (baseline = 3.47 +/- 0.18:1; MFAT = 3.33 +/- 0.14:1). CONCLUSION: The results suggest that reducing IMCL via 3-d consumption of a LFAT diet does not impair running performance lasting a little over 2 h (compared with 3-d consumption of a MFAT diet plus 1-d glycogen normalization), but that even short-term consumption of a LFAT diet may unfavorably alter serum lipids, even in healthy, endurance-trained runners. PMID- 18408609 TI - Effect of heat acclimation on sweat minerals. AB - PURPOSE: This study examined the impact of 10 d of exercise-heat acclimation on sweat mineral concentrations. METHODS: Eight male subjects walked on a treadmill at 3.5 mph, 4% grade for 100 continuous minutes or until rectal temperature reached 39.5 degrees C on 10 consecutive days in an environmental chamber set at 45 degrees C, 20% relative humidity. Arm sweat samples were collected during the first 30 min of exercise-heat stress on days 1 and 10 using a polyethylene arm glove. RESULTS: Final core temperature and HR values were significantly lower (P < 0.05) on day 10 versus day 1. Whole-body sweating rates increased by approximately 6% (P = 0.12). Sweat sodium concentration on day 10 (36.22 +/- 7.22 mM) was significantly lower than day 1 (54.49 +/- 16.18 mM) (P < 0.05). Sweat mineral concentrations of calcium (approximately 29%), copper (approximately 50%), and magnesium (approximately 43%) were also significantly lower on day 10 versus day 1 of heat acclimation (P < 0.05). A trend for lower sweat iron (approximately 75%; P = 0.07) and zinc (approximately 23%; P = 0.10) concentrations were observed from day 1 to day 10. The estimated hourly sweat mineral losses (arm concentration x whole-body sweat rate) were reduced for calcium (approximately 27%), copper (approximately 46%), and magnesium (approximately 42%) (P < 0.05), but not iron (75%) or zinc (approximately 16%) (P > 0.05), from day 1 to day 10. CONCLUSION: Exercise-heat acclimation conserves arm sweat mineral concentrations and possibly whole-body sweat losses of calcium, copper, and magnesium, and may reduce sweat iron and zinc concentrations. PMID- 18408610 TI - The effects of acute dopamine reuptake inhibition on performance. AB - INTRODUCTION: Acute bupropion (dopamine/noradrenaline reuptake inhibitor) administration significantly improved time trial performance and increased core temperature in the heat (30 degrees C). PURPOSE: The present study was performed to examine the effect of a dopaminergic reuptake inhibitor on exercise capacity and thermoregulation during prolonged exercise in temperate and warm conditions. METHODS: Eight healthy well-trained male cyclists participated in this study. Subjects ingested either placebo (PLA; 20 mg) or methylphenidate (MPH; Ritalin; 20 mg) 1 h before the start of exercise in temperate (18 degrees C) or warm (30 degrees C) conditions and cycled for 60 min at 55% Wmax, immediately followed by a time trial (TT; PLA18 and MPH18; PLA30 and MPH30) to measure exercise performance. RESULTS: MPH did not influence TT performance at 18 degrees C (P = 0.397). TT was completed 16% faster in MPH30 (38.1 +/- 6.4 min) than in PLA30 (45.4 +/- 7.3 min; P = 0.049). In the heat Tcore was significantly higher at rest (P = 0.009), and throughout the TT in MPH30 (P < 0.018), reaching values above 40 degrees C. Throughout MPH30, heart rates were significantly higher (P < 0.05). CONCLUSIONS: These results show that MPH has a clear ergogenic effect that was not apparent in 18 degrees C. The combination of a dopamine reuptake inhibitor and exercise in the heat clearly improved performance and caused hyperthermia without any change in the perception of effort or thermal stress compared with the PLA trial. This response may potentially increase the risk of developing heat illness during exercise in individuals taking drugs of this nature. PMID- 18408611 TI - Effects of different doses of caffeine on exercise responses in young children. AB - INTRODUCTION/PURPOSE: This study investigated the effects of three different doses of caffeine on physiological responses to exercise in young children. METHODS: Forty healthy children (20 boys and 20 girls) volunteered for a random, double-blind, counterbalanced study where they received either placebo (PL), 1 mg.kg(-1) (CAF-1), 3 mg.kg(-1) (CAF-3), or 5 mg.kg(-1) (CAF-5) caffeine, 60 min prior to preexercise measures, followed by cycle ergometer exercise at 25 W and then 60% VO2peak. During this time, heart rate (HR), oxygen consumption (VO2), and respiratory exchange ratio (RER) were measured continuously, while blood pressure (BP) was measured every 2 min. RESULTS: There were no significant gender x treatment interactions, and so the boys' and girls' data were combined. At preexercise, CAF-5 SBP and DBP were significantly (P < 0.05) higher than PL. Both CAF-1 and CAF-3 DBP were also significantly (P < 0.05) higher versus PL at preexercise. There were no treatment effects for exercise BP. At rest and during both exercise intensities, HR was significantly (P < 0.05) lower in CAF-3 (approximately 5 bpm) and CAF-5 (approximately 6 bpm) versus PL. There were no significant effects of CAF on metabolism (VO2 or RER) except for a lower RER in CAF-1 versus CAF-5 at 60% VO2peak. CONCLUSION: Low, mild, and moderate (1, 3, and 5 mg.kg(-1)) doses of caffeine have no effect on substrate use as reflected by RER. Further, caffeine intake resulted in an increase in BP and decrease in HR at preexercise and a slight decrease in HR with CAF-3 and CAF-5 versus PL during exercise. PMID- 18408612 TI - Recovery pattern of baroreflex sensitivity after exercise. AB - PURPOSE: To test the association between exercise mode and the recovery pattern of baroreflex sensitivity (BRS) after exercise. METHODS: The study population included healthy male subjects (N = 12, age: 31 +/- 3 yr). Four different interventions were performed in a randomized order: 1) aerobic exercise session on a bicycle ergometer, 2) light resistance exercise session, 3) heavy resistance exercise session, and 4) control intervention with no exercise. All interventions lasted 40 min. R-R intervals and continuous blood pressure were measured before (10 min) and 30-180 min after the interventions. BRSLF was calculated by the transfer function method from the low-frequency band (LF, 0.04-0.15 Hz) of the R R intervals and systolic blood pressure spectra. RESULTS: BRSLF had blunted until 30 min after aerobic and light resistance exercise (11.1 +/- 4.3 and 10.0 +/- 3.6 vs 17.5 +/- 7.0 ms.mm Hg(-1), P = 0.002 for both, compared with the control intervention, respectively). However, BRSLF was significantly blunted until 60 min after heavy resistance exercise (9.3 +/- 2.3 vs 15.1 +/- 4.7 ms.mm Hg(-1), P = 0.005, compared with the control intervention). The high-frequency power of R-R intervals (0.15-0.4 Hz) was significantly reduced, and the LF power of systolic blood pressure oscillation was significantly augmented 30 min after heavy resistance exercise (P < 0.01 for both), whereas both indices were restored to the control level by 30 min after aerobic and light resistance exercise. CONCLUSION: BRS after acute exercise is associated with exercise intensity, showing relatively rapid recovery after aerobic and light resistance exercise and delayed recovery after heavy resistance exercise. The delayed BRS pattern after heavy resistance exercise is regulated by delicate interplay between the withdrawal of vagal outflow and the probably increased sympathetic vasomotor tone documented by measurements of heart rate and blood pressure variability. PMID- 18408613 TI - Adaptations to short-term muscle unloading in young and aged men. AB - INTRODUCTION: The purpose of this investigation was to determine whether young (21.7 yr) and aged (68.5 yr) men experienced similar responses to 7 d of muscle unloading (N = 10 per group). METHODS: Unilateral lower limb suspension (ULLS) was used to impose muscle unloading of the knee extensors. To compare the effects of unloading on aged and young men, a repeated-measures factorial ANOVA was used to assess those effects on isometric strength, as well as strength, total work, and average power during isokinetic contractions conducted at 0.53, 1.05, and 2.09 rad.s(-1). RESULTS: Data showed that at slower speeds of movement, only a main effect of unloading was identified with young and aged men displaying similar and significant (P < 0.05) ULLS-induced decrements in strength, work, and power. The decrease in isometric strength correlated well with loss of electromyographic activity of contracting muscles (r = 0.79, P = 0.0002). At higher speeds of isokinetic contractions, not only was a main effect of age detected (young > aged), but it was also revealed that aged men, but not young men, experienced significant unloading-induced declines in muscle performance. Moreover, unloading resulted in a significant increase in plasma cortisol, a potent catabolic hormone, only among aged men. In contrast to other variables assessed, muscle endurance, quantified during 30 repetitions completed at 3.14 rad.s(-1), did not differ between age groups, nor was it altered by unloading. CONCLUSION: These findings suggest that young and aged men respond differently to muscle unloading, but in assessing muscle performance, these differences are manifested only during faster contractile velocities. PMID- 18408614 TI - Intermittent reloading attenuates muscle atrophy through modulating Akt/mTOR pathway. AB - PURPOSE: The aim of this study is to investigate the effects of intermittent reloading during hindlimb unloading (HU) on the changes in intracellular signaling pathways in skeletal muscle. METHODS: Male Wister rats were divided randomly into one of three experimental groups: 1) nonsuspended control, 2) HU for 7 d, and 3) HU with intermittent reloading (HU/IR) for 4 h.d(-1). After each experimental period, the antigravitational soleus muscle was analyzed. RESULTS: After 7 d of HU treatment, muscle fiber atrophy (decrease in relative muscle mass: 0.28 mg.g(-1) in the HU group vs 0.36 mg.g(-1) in the control group, P < 0.05; decrease in fiber CSA: 1682.6 microm2 in the HU group vs 2673.0 microm2 in the control group, P < 0.05) and a decrease in phosphorylation levels of anabolic signaling pathway (Akt and mTOR) were observed. Additionally, expressions of two types of muscle-specific E3 ubiquitin ligase mRNA (muscle atrophy F-box (MAFbx), and muscle ring finger 1 (MuRF1)) were upregulated during muscle atrophy. Increases in binding activities of nuclear factor kappa B (NFkappaB) were also determined. In contrast, IR treatment attenuated the muscle fiber atrophy (0.33 mg.g(-1) and 2067.5 microm2) and partially increased the phosphorylation levels of anabolic signaling molecules. Expression of MAFbx and MuRF1 mRNA were returned to the control level, and binding activities of nuclear NFkappaB was suppressed with the effects of IR. CONCLUSION: These results suggest that IR-induced attenuation of skeletal muscle atrophy is achieved by the synergy between increased anabolic and decreased catabolic signaling mechanisms. PMID- 18408615 TI - Dose-dependent hepatic response to subchronic administration of nandrolone decanoate. AB - BACKGROUND: Androgenic anabolic steroids (AAS) are synthetic hormone derivatives of testosterone and are mainly used to enhance athletic performance and muscle mass, but medical applications also have been described. Short- and long-term side effects have been demonstrated in many organs, but the liver adverse effects are the most common and serious ones associated with AAS use. However, these effects have been supported by few clinical and experimental studies. OBJECTIVE: To evaluate the hepatic function and structure after 5 wk of nandrolone decanoate administration at three different doses. METHODS: Twenty-seven adult male Wistar rats were randomly assigned to the following groups: control, clinical, intermediate, and suprapharmacological doses of nandrolone decanoate during 5 wk. RESULTS: The biochemical studies showed that nandrolone decanoate administration leads to a dose-dependent increase in serum levels of the aspartate aminotransferase (AST) (P < 0.05), alanine aminotransferase (ALT) (P < 0.01), and alkaline phosphatase (ALP) (P < 0.001), as well as a significant decrease in total proteins (P < 0.01), bilirubin (P < 0.05), total cholesterol and fractions (P < 0.05), and triglycerides (P < 0.05). Although a significant statistical difference was found for AST, ALT, and ALP when compared with the control group, their values remained within the normal range. The number of Kupffer cells was increased in the liver parenchyma (P < 0.05), and the content of collagen was increased in the central lobular vein wall, in the hepatic parenchyma, and in the portal space (P < 0.05). CONCLUSIONS: These results suggest that subchronic treatment with nandrolone decanoate, mainly administered at higher-than-clinical doses, are potentially deleterious to the liver, leading to incipient fibrosis. PMID- 18408616 TI - Physical activity among adolescents in New South Wales (Australia): 1997 and 2004. AB - PURPOSE: To examine secular trends, seasonal and socioeconomic differences in physical activity participation among Australian adolescents in 1997 and 2004. METHODS: Repeat cross-sectional school survey, conducted in 1997 and 2004. School students were randomly selected from grades 8 and 10 in New South Wales Australia (1997, N = 2026; 2004 N = 1771). Time spent in moderate-to-vigorous physical activity (MVPA) was assessed by self-reported participation in organized and nonorganized physical activity separately for summer and winter by sex, grade, and socioeconomic status (SES). RESULTS: Between survey periods, the proportion of students reporting > or = 60 min.d(-1) of MVPA increased, except among grade 10 boys during winter. MVPA (min.d(-1)) increased among all groups during summer school terms, during winter school terms, MVPA was relatively stable between surveys for grade 8 students but inconsistent among grade 10 students. Organized MVPA increased among all groups in both summer (19-49 min.d(-1)) and winter (7-21 min.d(-1)) school terms. Participation in nonorganized MVPA increased only during summer school terms (4-32 min.d(-1)) and decreased during winter school terms (7 17 min.d(-1)). MVPA increased consistently among students in the high-SES group compared with other SES groups, independent of season, and in some cases MVPA decreased in low-SES groups during winter school terms. CONCLUSION: Overall, the patterns of change were similar for boys and girls, with substantial increases in summer school terms and insubstantial changes during winter school terms (with the exception of a decrease among grade 10 boys). There was a direct association between change in participation in MVPA and SES, indicating that interventions are required to focus on the needs of less-advantaged students. PMID- 18408617 TI - Physical activity, sedentary index, and mental disorders in the SUN cohort study. AB - PURPOSE: There is evidence to suggest a beneficial effect of physical activity on several mental disorders. METHODS: The study aim was to assess the association between low physical activity during leisure time (or sedentary lifestyles) and the incidence of mental disorders in 10,381 participants, from a Spanish dynamic prospective cohort of university graduates followed up for 6 yr (the SUN study). The baseline assessment included a validated questionnaire on physical activity during leisure-time and sedentary activities. A subject was classified as an incident case of mental disorder if he or she reported a physician diagnosis of depression, anxiety or stress, and/or the use of antidepressant medication or tranquilizers in at least one of the follow-up questionnaires. RESULTS: The odds ratios (OR) and 95% confidence intervals (CI) of a mental disorder for successive levels of leisure-time physical activity were 1 (reference), 1.00 (0.81, 1.23), 0.99 (0.81, 1.21), 0.72 (0.58, 0.89), and 0.81 (0.65, 1.00) (P for trend: < 0.01). The OR for subjects who spent more than 42 h.wk(-1) watching television and/or using the computer was 1.31 (95% CI = 1.01, 1.68) as compared with those spending less than 10.5 h.wk(-1). CONCLUSION: Our findings suggest a joint association of leisure-time physical activity and sedentary behavior on the incidence of mental disorders. PMID- 18408618 TI - Depressive symptoms and physical activity in adolescent girls. AB - PURPOSE: To evaluate the relationship between depressive symptoms and physical activity in a geographically and ethnically diverse sample of sixth-grade adolescent girls. METHODS: The Trial of Activity for Adolescent Girls (TAAG) baseline measurement included a random sample (N = 1721) of sixth-grade girls in 36 schools at six field sites. Measurements were accelerometry and the 3-d Physical Activity Recall (3DPAR) for physical activity, and the Center for Epidemiological Studies-Depression scale (CES-D) for depressive symptoms. RESULTS: Girls with complete data (N = 1397), mean age 12 yr, had an average CES D score of 14.7 (SD = 9.25) and engaged in an average of about 460 min of sedentary activity, < 24 min of moderate to vigorous physical activity (MVPA), and < 6 min of vigorous physical activity (VPA) in an 18-h day. Thirty-minute segments of MVPA ranged in number from 3.9 to 1.2, and METS for these segments ranged from > 3.0 to > 6.5. Mixed-model regression indicated no relationship between depressive symptoms and physical activity; however, a significant but modest inverse relationship between sedentary activity and depressive symptoms was observed. CONCLUSION: A sufficient sample size, standardized procedures, and validated instruments characterized this study; however, a relationship between depressive symptoms and physical activity was not observed for sixth-grade girls from diverse geographic locations. The average CES-D score was lower than is considered clinically meaningful for either adolescents or adults, and MET minutes of sedentary activity were high. This combination of data may be different from other studies and could have contributed to the unexpected finding. This unexpected finding is informative, however, because it shows the need for additional research that includes a wider range of possible combinations of data, especially with young adolescent girls. PMID- 18408619 TI - Exercise preconditioning protects against doxorubicin-induced cardiac dysfunction. AB - The clinical use of the chemotherapeutic drug doxorubicin (DOX) is limited due to a dose-dependent cardiotoxicity. Evidence is mounting that exercise protects against DOX-related cardiac dysfunction, and as such, it may be possible that prior endurance training promotes defense against DOX cardiotoxicity. PURPOSE: To examine the effects of exercise preconditioning on acute DOX-induced cardiotoxicity, and to determine whether any observed cardioprotection was associated with myosin heavy chain (MHC) isoform alterations. METHODS: Male Sprague-Dawley rats trained on a motorized treadmill, had access to voluntary running wheels, or remained sedentary for 10 wk prior to being injected with either saline or 10 mg.kg(-1) DOX. Left ventricular function was then assessed in vivo using transthoracic echocardiography and ex vivo using the isolated working heart at 5 and 10 d after injection. Additionally, left ventricular MHC isoform expression was analyzed as a possible mechanism to explain exercise-induced cardioprotection. RESULTS: DOX treatment promoted significant in vivo and ex vivo cardiac dysfunction at 5 and 10 d after injection in sedentary animals, and this dysfunction was associated with an upregulation of the beta-MHC isoform. Exercise preconditioning protected against DOX-induced cardiac dysfunction at 5 and 10 d after injection by attenuating beta-MHC upregulation. CONCLUSION: Endurance training prior to DOX treatment protects against acute DOX cardiotoxicity for up to 10 d, and this protection can potentially be explained by a preservation of MHC isoform distribution. PMID- 18408620 TI - Curing the cure: utilizing exercise to limit cardiotoxicity. PMID- 18408621 TI - Ventilatory efficiency and resting hemodynamics in hypertrophic cardiomyopathy. AB - PURPOSE: In patients with systolic heart failure, the ability of cardiopulmonary exercise testing (CPX) variables to reflect pathophysiology is well established. The relationship between CPX and pathophysiology has, however, not been thoroughly investigated in patients with nonobstructive hypertrophic cardiomyopathy (NHCM). The objective of this study was to assess the ability of CPX variables to reflect resting hemodynamics in patients with nonobstructive hypertrophic cardiomyopathy NHCM. METHODS: We performed CPX and right heart catheterization on 83 subjects with NHCM (51 male/32 female, mean age = 38 +/- 10 yr, NYHA I-III mean = 1.7). Peak oxygen consumption ( O2) and minute ventilation/carbon dioxide ratio (V E/VCO2) at peak exercise were compared to resting hemodynamics including pulmonary artery systolic, diastolic and mean pressures (PASP, PADP and MPAP), and pulmonary capillary wedge pressure (PCWP). RESULTS: Elevations in PCWP (> or = 15 mm Hg), PASP (> or =30 and > or = 40 mm Hg), PADP (> 15 mm Hg) and MPAP (> or = 20 mm Hg) were detected in 22, 33, 10, and 23% of subjects, respectively. Peak V E/VCO2 (positive correlation) and peak VO2 (negative correlation) correlated modestly with all pressure measurements (r = 0.33-0.51, P < 0.01 for all measurements). By receiver operating curve analysis, a V E/VCO2 >35.5 exhibited the best diagnostic accuracy with a curve areas of 0.81 for PAP > or = 30 mm Hg (sensitivity/specificity = 86%/67%), 0.87 for PAP > or = 40 mm Hg (77%/100%), 0.86 for MPAP > 20 mm Hg (83%/79%), and 0.84 for PCWP > or = 15 mm Hg (80%/76%). CONCLUSIONS: CPX can accurately identify abnormal resting hemodynamics in patients with NHCM. Further testing of this modality in other forms of diastolic dysfunction may be warranted. PMID- 18408622 TI - Electrocardiograms in athletes: interpretation and diagnostic accuracy. AB - PURPOSE: Electrocardiography (ECG) has been proposed as a method to enhance the ability of the preparticipation examination (PPE) to detect underlying cardiac conditions that can lead to sudden cardiac death (SCD) in young athletes. METHODS AND RESULTS: We conducted a Medline review of the published medical literature, using the key terms of cardiovascular screening of athletes, ECG in athletes, SCD in athletes, and ECG in specific cardiac disease states: hypertrophic cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, dilated cardiomyopathy, myocarditis, long QT syndrome, Brugada syndrome, coronary artery anomalies, myocardial bridging, aortic stenosis, mitral valve prolapse, and Marfan syndrome. ECG seems to increase the sensitivity of the PPE from 2.5-6% to 50-95%. Overall sensitivity appears to be about 50%; false-positive rates can be as high as 40%, and there is at least a 4-5% false-negative rate. In Europe, ECG based screening programs have been associated with a decline in the SCD rate in young athletes, but similar programs are currently not recommended in the United States for many reasons: lack of randomized trial data; cost of screening; lack of a clear standard for ECG interpretation in the athlete; the likelihood that asymptomatic athletes with underlying lethal conditions might differ significantly from symptomatic individuals with the same conditions; and concern that ECG screening might actually increase the death rate, via treatment-related procedural complications. CONCLUSIONS: Although some authorities advocate the use of ECG screening of young athletes, further studies are required to define what constitutes a normal ECG in athletes, and to determine whether ECG-based screening protocols truly are superior, not only in finding disease, but also saving lives. For those who either choose ECG-based screening or interpret ECG in athletes, we propose a simple interpretation scheme and decision tree. PMID- 18408623 TI - HDL atheroprotection by aerobic exercise training in type 2 diabetes mellitus. AB - PURPOSE: In this study we analyzed the role played by aerobic exercise training in the plasma lipoprotein profile, prebeta 1-HDL concentration, and in the in vitro HDL3 ability to remove cholesterol from macrophages and inhibit LDL oxidation in type 2 diabetes mellitus (DM) patients and control subjects, in the fasting and postprandial states. METHODS: Healthy controls (HTC, N = 11; 1 M/10 F) and subjects with type 2 diabetes mellitus (DMT, N = 11; 3M/8F) were engaged in a 4-month aerobic training program, and compared with a group of sedentary subjects with type 2 diabetes mellitus (DMS, N = 10; 4 M/6 F). All groups were submitted to an oral fat load test to analyze all parameters, both at the beginning of the investigation protocol (basal) and at the end of the study period (final). RESULTS: Exercising did not modify body weight, BMI, plasma concentrations of total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides (TG), glucose, insulin, or HOMA-IR, but it reduced the waist circumference. The HDL3 composition did not change, and its ability to remove cell cholesterol was unaltered by aerobic training. In DMT but not in HTC, aerobic training improved 15% the HDL3 protective effect against LDL maximal oxidation rate in the fasting state, and reduced 24% the plasma prebeta 1-HDL concentration in the postprandial state, suggesting an enhanced prebeta 1-HDL conversion into larger, more mature HDL particles. In this regard, regular aerobic exercise enriched HDL2 with TG in the fasting and postprandial states in HTC and in the fasting phase in DMT. CONCLUSION: Our results show that aerobic exercise training in diabetes mellitus improves the HDL efficiency against LDL oxidation and favors HDL maturation. These findings were independent of changes in insulin resistance and of the rise of plasma HDL cholesterol concentration. PMID- 18408624 TI - BDNF gene is a genetic risk factor for schizophrenia and is related to the chlorpromazine-induced extrapyramidal syndrome in the Chinese population. AB - BACKGROUND: Brain-derived neurotrophic factor (BDNF) belongs to a family of the neurotrophin, which plays important roles in the neurodevelopment of dopaminergic related systems and interacts with meso-limbic dopaminergic systems involved in the therapeutic response to antipsychotics. Functional experiments have suggested that BDNF may be involved in the etiology of schizophrenia. METHODS AND RESULTS: In this study, we genotyped two important functional polymorphisms in the BDNF gene using a sample of Han Chinese patients consisting of 340 schizophrenic patients and 343 healthy controls. We found a statistical difference in the 232 bp allele distribution of the BDNF gene (GT)n dinucleotide repeat polymorphism between the schizophrenic patients and controls. In early onset patients, the 234 bp allele had a risk role. For the chlorpromazine-induced extrapyramidal syndrome, the 230-bp allele and the 234-bp allele acted in opposite directions, that is, patients with the 230-bp allele of the (GT)n polymorphism exhibited a lower degree of induced extrapyramidal syndrome. Haplotype-based analysis also revealed a very important risk haplotype (P=0.0000226546). CONCLUSION: These findings suggest that BDNF plays an important role in the susceptibility to schizophrenia and that the (GT)n repeat polymorphism of the BDNF gene may be an independent contributor to the chlorpromazine treatment-sensitive form of schizophrenia. PMID- 18408625 TI - [Intervention or procedure? What's at stake for interventional radiology]. PMID- 18408626 TI - [Impact of CT on negative appendectomy and appendiceal perforation rates]. PMID- 18408628 TI - [MRI of the post-traumatic penis]. AB - Penile trauma, though rare, requires specialized management. Fibrotic sequelae from lesions of the tunica albuginea and corpora cavernosa must be prevented given their impact of the erectile function. Knowledge of penile anatomy enables acquisition and interpretation of penile MRI, preferably performed in the first few hours following injury. Tear of the tunica albuginea must be identified since it requires surgical management. Other lesions may be present at the acute (hematoma) or chronic (fibrosis, arteriocavernous fistula) phases. MRI, non invasive and painless, is the imaging modality of choice in the multiplanar evaluation of traumatic injuries of the penis. PMID- 18408627 TI - [High resolution MRI of carotid atherosclerosis: looking beyond the arterial lumen]. AB - Despite advances in diagnosis and treatment, atherosclerosis remains the second cause of death in the world. Due to technical advances, high resolution MRI (HR MRI) allows depiction of the wall of cervical arteries, especially carotid atherosclerosis. HR-MRI allows visualization of the different components of atherosclerosis: necrotic lipid core, intraplaque hemorrhage, calcifications and fibrous cap. Global plaque volume as well as the volumes of individual plaque components can be calculated. Atherosclerotic plaque structure analysis, along with stenosis measurement, contribute to the stratification of the stroke risk. HR-MRI may also be used to assess treatment efficacy aimed at stabilizing or reducing plaque progression. Beyond the arterial lumen, direct evaluation of vessel wall should modify the management of atherosclerosis in the years to come. PMID- 18408629 TI - [Splanchnic vein aneurysms: a report of 13 cases]. AB - PURPOSE: Splanchnic vein aneurysms are a rare and poorly known entity. From a review of 13 cases and reports from the literature, we have defined the imaging features and modalities for their diagnosis and management. PATIENTS AND METHODS: Retrospective study of 13 patients (7 females, 6 males, mean age of 53.5 years) who presented between 1993 and 2005 with splanchnic vein aneurysms. All patient charts were reviewed to collect and analyze epidemiological data, underlying etiologies, imaging work-up for diagnosis and management. Aneurysms were defined as fusiform or saccular enlargement of the portal venous vein or its branches, superior mesenteric vein and splenic vein. RESULTS: A total of 16 splanchnic vein aneurysms were identified. Four patients had portal venous hypertension. The imaging work-up was variable and included mainly non-invasive modalities (US: 8 patients; CTA: 8 patients and MRA: 9 patients) and conventional angiography in 2 patients. Eight aneurysms involved the portal vein (50%), 5 the splenic vein (31.3%), and 3 the superior mesenteric vein (18.7%). None of the patients underwent surgical or endovascular management. Imaging follow-up was performed in 12 patients, with no interval change over time with up to 10 years follow-up. CONCLUSION: In our series, the portal vein was most frequently involved. Diagnosis was exclusively based on imaging and management was limited to follow up. PMID- 18408630 TI - [Evaluation of CT guided cervical epidural injections in patients with mechanical cervicobrachial neuralgia]. AB - PURPOSE: Prospective study evaluating the efficacy and tolerability of CT guided cervical epidural injections for mechanical cervicobrachial neuralgia. MATERIALS AND METHODS: Between July 2003 and August 2004, 34 patients (16 females, 18 males) with mean age of 44.5 years were enrolled. Enrollment criteria included mechanical cervicobrachial neuralgia of more than 1 month's duration, refractory to medical management, with good correlation between clinical and imaging findings. An evaluation using the Neck Pain Disability Score (NPDS) and associated Secondary criteria was performed by a single observer at day 0, day 30 and day 90. The injections were performed under CT guidance. RESULTS: The symptomatic efficacy of CT guided injections at 3 months was 45% using the NPDS, 47.1% using an analog pain scale, and 58.8% as graded by patients (effective/very effective). Tolerability was excellent (55.9%), moderate (41.2%) and poor (2.9%). CONCLUSION: CT guided cervical epidural injections appear effective and well tolerated. A blinded randomized study could validate its value. However, a study comparing steroids and placebo raises ethical concerns. PMID- 18408631 TI - [Value of double reading of whole body CT in polytrauma patients]. AB - PURPOSE: To assess the value of standard double reading of whole body CT in the management of polytrauma patients. MATERIALS AND METHODS: Prospective study between January and July 2005. Two senior radiologists with expertise in trauma imaging, blinded to clinical findings, reviewed 105 initial CT examinations of polytrauma patients. These examinations had initially been interpreted by the on call radiologist. The second interpretations were performed within 12 hours of admission, and were considered the gold standard. RESULTS: A total of 105 patients were included with 82 males (78%) and 23 females (22%), aged between 2 and 83 years. The level of admission was graded III (n=64), II (n=30) and I (n=11). The second reading identified 3 lesions that were not initially described, each requiring a change in management, including splenic rupture (n=1), thoracic spine fracture (n=1) and epidural hematoma (n=1), with no unfavorable impact on mortality. Additional errors in the initial interpretation were identified: peripheral fractures (n=38), chest (n=36), brain (n=31), abdominal (n=28), spine (n=19) and maxillofacial (17) lesions and contrast extravasation (n=6). CONCLUSION: Based on the large number and severity of some lesions missed at initial interpretation of whole body CT of polytrauma patients, we recommend standard double reading of these examinations. PMID- 18408632 TI - [Harmonic sonography of rotator cuff tendons: are cleavage tears visible at last?]. AB - PURPOSE: For some, cleavage tears remain a pitfall of sonography (US). The purpose of this study is to demonstrate the visibility of intratendinous tears of the supraspinatus and infraspinatus tendons and describe their imaging features on tissue harmonic US. MATERIALS AND METHODS: Prospective study of 52 patients with suspected cuff pathology who underwent US prior to CT-arthrography, CT bursography or MR arthrography. The US examinations were performed using 7-15 MHz transducers with tissue harmonic mode (pulse subtraction). An intratendinous tear was suggested by the presence of a hypoechoic intratendinous line, extending from a partial or full thickness tear. Results from US were correlated to contrast material enhanced CT or MR findings. RESULTS: Ten cases of cleavage tears were detected on US compared to 18 on arthrographic examinations. False negative results occurred in poor US candidates (n=4), excessively retracted tendons (n=2) or postsurgical cuffs (n=2). Sensitivity was 55% and specificity was 94%, with PPV of 83% and NPV of 80% for the detection of intratendinous tear. The accuracy of US was lower for intratendinous tear associated with full thickness tears (5/11) compared to intratendinous tears associated with partial thickness tears (5/6). CONCLUSION: We demonstrate that cleavage tears are now visible on US using high-frequency transducers and tissue harmonic mode. However, the sensitivity remains too low, especially in patients with full thickness tear, postsurgical patients and patients that are poor candidates to US. PMID- 18408633 TI - [MRI features of mycetoma of the foot: report of two cases and review of the literature]. PMID- 18408634 TI - [Pseudoaneurysm of the cystic artery complicating calculous cholecystitis: a case report]. PMID- 18408635 TI - [Decompression sickness of the spinal cord: MRI features]. PMID- 18408636 TI - [US diagnosis of benign hypertrophic gastritis in a child]. PMID- 18408637 TI - [Answer to February e-quid: diagnosis of a duodenal duplication]. PMID- 18408639 TI - [Imaging and PET-CT of adult and childhood lymphoma]. AB - Malignant lymphomas are lymphoproliferative disorders arising in both lymphoid tissue and non-lymphoid organ systems. Treatment rarely is surgical, and currently relies on a combination of chemotherapy and radiation therapy. The role of imaging is to determine the spread of the disease, to identify targets and to assess therapeutic response. Imaging techniques mainly use morphological criteria, and may underestimate infiltrative disease, as observed in bones. The frequent presence of residual masses after treatment usually prevents classification of patients as complete response. Over time, positron emission tomography (PET) with F18-fluorodeoxyglucose (FDG) has become a prominent part of the workup at diagnosis and during follow-up. Recently, PET has been integrated in the revised response criteria for malignant lymphoma. PMID- 18408640 TI - [Imaging and PET/CT of lung cancer]. AB - Lung cancer is one of the most frequently occurring cancer in the world. Imaging plays a critical role for screening, diagnosing, staging, and following patients. Although morphologic imaging such as chest X-ray and CT are still useful for these purpose, major limitations occur in the proper evaluation of diagnosing and staging. Metabolic imaging using PET significantly increases the accuracy of staging. This paper will review the role of imaging in patients suspected or diagnosed with lung cancer. PMID- 18408641 TI - [Imaging and PET-CT of head and neck cancers]. PMID- 18408642 TI - [Imaging and PET/CT evaluation of GI tract cancers]. AB - Imaging plays a pivotal role in the management of GI tract cancers for diagnosis, characterization, locoregional staging, metastatic work-up and follow-up during and after curative or palliative treatment. The imaging protocols should be optimized and reproducible because of their impact on therapy. Thoracic, abdominal and pelvic CT is the cornerstone of the imaging work-up, optimized and tailored to the specific GI segment involved, requiring good GI tract distension. Image interpretation of native axial and reformatted multiplanar images is routinely performed. In specific cases, additional targeted imaging with US or MRI or whole body imaging with PET/CT or MRI may be valuable. PET/CT is a complement to morphological imaging. PET allows detection of lesions otherwise undetected on morphological imaging, usually due to poor contrast with surrounding tissues, and characterization of known lesions. PET/CT is best used as an integral part of a comprehensive imaging work-up. Radiologist and nuclear medicine specialist provide complementary information. Each must be familiar with the clinical questions at hand and related stakes, and advantages and limitations of each modality to optimize treatment as part of a multidisciplinary management approach. PMID- 18408643 TI - Imaging and PET-PET/CT imaging. AB - PET-CT has grown because the lack of anatomic landmarks in PET makes "hardware fusion" to anatomic cross-sectional data extremely useful. Addition of CT to PET improves specificity, but also sensitivity, and adding PET to CT adds sensitivity and specificity in tumor imaging. The synergistic advantage of adding CT is that the attenuation correction needed for PET data can also be derived from the CT data. This makes PET-CT 25-30% faster than PET alone, leading to higher patient throughput and a more comfortable examination for patients typically lasting 20 minutes or less. FDG-PET-CT appears to provide relevant information in the staging and therapy monitoring of many tumors, such as lung carcinoma, colorectal cancer, lymphoma, gynaecological cancers, melanoma and many others, with the notable exception of prostatic cancer. For this cancer, choline derivatives may possibly become useful radiopharmaceuticals. The published literature on the applications of FDG-PET-CT in oncology is still limited but several well-designed studies have demonstrated the benefits of PET-CT. PMID- 18408644 TI - Brain stimulation, alternative medicine, and impulsivity and compulsivity. PMID- 18408645 TI - Self-mutilation: a symptom of psychiatric disorder or a nosological entity with its own characteristics? PMID- 18408646 TI - Do dopamine partial agonists have partial efficacy as antipsychotics? PMID- 18408647 TI - Brain plasticity and brain stimulation in neuropsychiatry: toward individualized medicine. PMID- 18408648 TI - Systematic reviews of clinical trials of acupuncture as treatment for depression: how systematic and accurate are they? PMID- 18408649 TI - Development of tics in a thirteen-year-old male following atomoxetine use. AB - Tics and Tourette syndrome are common comorbidities of patients diagnosed with attention-deficit/hyperactivity disorder (ADHD). One of the mainstay pharmacologic therapies for ADHD has been stimulants. However, this class of drugs has been associated with tic exacerbations, thus limiting their utility in this patients subgroup. Atomoxetine has been explored as an alternative treatment as one of the few non-stimulants available to treat ADHD. Early data identifies atomoxetine's influence on Tourette symptomatology to be not merely equivocal but potentially suppressive in the manifestation of tics. There are, however, case studies describing patients experiencing recurrences of tics following treatment with atomoxetine. We present a unique case of a patient, without any prior history of a movement disorder, who developed tics following a single dose of atomoxetine that did not improve until interventional therapy was initiated. PMID- 18408650 TI - Neuropsychological characteristics and personality traits in pathological gambling. AB - INTRODUCTION: Pathological gambling disorder (PG) has been associated with fronto temporal dysfunction and maladaptive personality traits, such as impulsivity and novelty seeking. The purpose of this study was to examine the predictive variance of neuropsychological and personality characteristics in PG. METHODS: Persons with PG (n=25) and a comparison group (n=34) were administered a battery of neuropsychological tests, the Temperament and Character Inventory, and the Barratt Impulsiveness Scale. Subjects with PG had evidence of fronto-temporal dysfunction as assessed by the Stroop, Wisconsin Card Sorting Test-64, Wechsler Adult Intelligence Scale Letter-Number Sequencing, Controlled Oral Word Association Test, and Boston Diagnostic Aphasia Examination Animal Naming Test. RESULTS: Subjects with PG also had impaired decision making on the Iowa Gambling Task. PG subjects had elevated levels of impulsivity, novelty seeking, and harm avoidance, and lower levels of self-directedness and cooperativeness. Logistic regression analyses indicated that neuropsychological variables did not add significant incremental variance over personality traits in predicting PG (Block chi-square=5.19, P=.074), while personality variables added significant incremental variance over neuropsychological traits in predicting PG (Block chi square=25.13, P<.001). CONCLUSION: These results suggest that personality traits are better predictors than neuropsychological characteristics of whether someone has PG. PMID- 18408651 TI - The prevalence of body dysmorphic disorder in the United States adult population. AB - OBJECTIVE: In clinical samples, body dysmorphic disorder (BDD) is associated with substantial suffering and reduced quality of life. Limited surveys report widely varying prevalence estimates. To better establish the prevalence of BDD, we conducted a United States nationwide prevalence survey. METHOD: We conducted a random sample national household telephone survey in the spring and summer of 2004 and interviewed 2,513 adults, of whom 2,048 qualified for the BDD-module administration. The computer-assisted, structured interviews, conducted by trained lay interviewers, addressed Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for BDD, along with information regarding several impulse-control disorders and the respondents' financial and demographic data. RESULTS: The rate of response was 56.3%, which compared favorably with rates in federal national health surveys. The cooperation rate was 97.6%. Respondents included a higher percentage of women and people >55 years of age than in the US adult population, and a lower percentage of Hispanics. The estimated point prevalence of DSM-IV BDD among respondents was 2.4% (49/2,048) (by gender: 2.5% for women, 2.2% for men), exceeding the prevalence of schizophrenia and bipolar disorder type I and about that of generalized anxiety disorder. BDD prevalence decreased after 44 years of age, and a larger proportion of BDD respondents were never married. Of those meeting DSM-IV criteria for BDD, 90% (45/49) met the DSM-IV distress criterion, and 51% (25/49) met the interference-with-functioning criterion. CONCLUSION: A study using clinically valid interviews is needed to evaluate these results. Such studies could inform treatment by documenting rates of seeking treatment from various sources, suicide attempt rates, and the prevalence of comorbid conditions. PMID- 18408652 TI - Rage attacks and aggressive symptoms in Japanese adolescents with tourette syndrome. AB - OBJECTIVE: This study was conducted to explore possible causes of rage attacks as well as clinically significant aggressive symptoms in Japanese adolescents with Tourette syndrome (TS). METHODS: The subjects included 29 adolescents (23 males, 6 females; mean age: 13.5+/-3.7 years). Eighteen subjects (62.1%) were diagnosed with TS only, 11 (37.9%) with TS and comorbidities, including attention deficit/hyperactivity disorder and obsessive-compulsive disorder. Parents completed the Child Behavior Checklist. Clinically significant aggressive symptoms were assessed using two pilot tools, the Rage Screen and Questionnaire and the Clinical Rating of Aggression. RESULTS: Thirteen subjects (44.8%) were judged to have clinically significant aggressive symptoms, according to the Clinical Rating of Aggression. Twelve met criteria for recurrent rage attacks, according to the Rage Screen and Questionnaire. Between the 13 aggressive and 16 non-aggressive subjects, no significant differences were found in age, gender, psychiatric comorbidities, or concurrent medication. Child Behavior Checklist ratings to compare 11 aggressive and 12 non-aggressive subjects <16 years of age revealed elevated t-test scores on the anxious/depressed, thought problems, aggressive, internalizing, externalizing subscales, and total scale in the aggressive group versus the non-aggressive group. CONCLUSION: Rage attacks and clinically significant aggressive symptoms are common problems in Japanese TS youth. Psychiatric morbidity appears associated with impulsive-aggressive symptoms. Treatment implications from these findings need to be explored further. PMID- 18408653 TI - Bridging the clinical gap: managing patients with co-occurring mood, anxiety, and alcohol use disorders. Introduction. PMID- 18408654 TI - Managing Parkinson's disease with continuous dopaminergic stimulation. AB - The pathophysiology of Parkinson's disease is marked by the loss of dopaminergic neurons, which leads to striatal dopaminergic deficiency. This causes resting tremor, hypokinesia, rigidity, bradykinesia, and loss of postural reflexes. Most current treatments for Parkinson's disease aim to restore striatal dopamine signaling by increasing the supply of dopamine with oral levodopa (L-dopa), stimulating dopamine receptors directly using dopamine agonists, or inhibiting the reuptake of endogenous dopamine. L-dopa is standard therapy for patients with Parkinson's disease. However, with continued treatment and disease progression, the response to oral dopaminergic drugs becomes unstable and motor fluctuations emerge, including off periods and dyskinesia. Direct duodenal-administered infusible L-dopa/carbidopa is effective for the management of refractory motor fluctuations in some patient populations. However, enteral infusions cannot mimic the function of the normal dopaminergic brain, and around-the-clock constant-rate administration carries the risk of causing refractory off periods associated with severe immobility and hyperpyrexia. Subthalamic nucleus (STN) deep brain stimulation (DBS) is also a promising treatment. DBS passes a high-frequency electrical current into the target area, mimicking the effect of lesioning the stimulated area. However, this treatment requires invasive surgery and is appropriate for a limited segment of the patient population. This supplement provides a rationale for the use of continuous dopaminergic receptor stimulation and offers guidelines on the individualization of treatment decisions, with special focus on continuous L-dopa infusion and STN DBS. Erik Wolters, MD, PhD, offers an introduction to the impact of continuous L-dopa infusion. Andrew J. Lees, MD, FRCP, provides an overview of the physiologic response to L-dopa and reviews clinical pharmacologic studies of intravenous and intraduodenal L-dopa. Jens Volkmann, MD, discusses selection criteria for STN DBS and duodenal L dopa/carbidopa infusion. Teus van Laar, MD, PhD, and Ad Hovestadt, MD, discuss the first data from a Dutch cohort study of duodenal L-dopa/carbidopa. PMID- 18408655 TI - NM23 as a prognostic biomarker in ovarian serous carcinoma. AB - The nm23 gene is a reported metastasis suppressor gene. Recent studies have shown that its expression has tissue specificity. The role of nm23 in human ovarian cancer is still controversial. This study examines the prognostic significance of nm23 expression in patients with serous ovarian carcinoma. Following comparative proteomics in 13 fresh frozen ovarian serous cancer tissues with other histological types of ovarian cancers, validation was performed using immunohistochemistry on clinically well-designed 73 ovarian serous carcinoma microarray samples that were retrieved from ovarian cancer patients from 1990 to 2003. Statistical analysis of the results was performed using chi(2) test, Cox proportional regression, the Kaplan-Meier method and log-rank test. We found that the expression of nm23 inversely correlated with peritoneal seeding (P=0.009). However, strong nm23 expression was associated with mortality in patients with ovarian carcinoma in univariate analysis (P=0.04). Poor prognostic factors of disease-free survival included tumor residue more than 2 cm (P=0.02), bilaterality (P=0.01) and peritoneal seeding (P<0.01), whereas poor prognostic factors affecting overall survival included peritoneal seeding (P=0.05). In Kaplan-Meier analysis, strong nm23 immunoreactivity correlates with poor overall survival (P=0.04) but not with poor disease-free survival. In conclusion, overexpression of nm23 is independently associated with decreased overall survival in patients with ovarian carcinoma and also significantly correlates with mortality. Nm23 may have a biological function that leads to poor clinical outcomes in ovarian carcinoma. PMID- 18408656 TI - Expression of VEGF-A/C, VEGF-R2, PDGF-alpha/beta, c-kit, EGFR, Her-2/Neu, Mcl-1 and Bmi-1 in Merkel cell carcinoma. AB - Merkel cell carcinoma is a rare but very aggressive tumor of the skin. With current treatment options, Merkel cell carcinoma is associated with a high incidence of recurrence and metastasis. Targeted anticancer therapies such as receptor tyrosine kinase inhibitors and antisense oligonucleotides have been found to be a promising new type of treatment for various types of cancer. To evaluate whether the use of targeted therapies is a possible treatment option in Merkel cell carcinoma, we determined the expression of the target molecules c kit, Mcl-1, Bmi-1, vascular endothelial growth factor (VEGF)-A, VEGF-C, VEGF receptor 2 (VEGF-R2), platelet-derived growth factor (PDGF)-alpha, PDGF-beta, epidermal growth factor receptor (EGFR) and Her-2/Neu in a tissue microarray of 32 samples of 29 patients with Merkel cell carcinoma. C-kit-positive samples were analyzed for mutations in exons 9 and 11. The tissue microarray was stained immunohistochemically with antibodies directed against the above-mentioned proteins, and an immunoreactivity score was calculated. DNA was extracted from c kit-positive samples and was analyzed for exon 9 and 11 mutations using direct DNA sequencing. We found that c-kit (7%), Mcl-1 (88%), Bmi-1 (78%), VEGF-A (91%), VEGF-C (75%) VEGF-R2 (88%), PDGF-alpha (72%) and PDGF-beta (13%) were expressed in Merkel cell carcinomas. All samples showed a lack of EGFR and Her-2/Neu expression. Analysis of c-kit revealed no mutations. As VEGF-A, VEGF-C, VEGF-R2, PDGFs and c-kit are targets of new cytostatic agents used in the treatment of other cancers, inhibition by a multitargeted chemotherapy could be a very promising treatment option. High expression of Bmi-1 and Mcl-1 warrants further studies on the use of antisense oligonucleotides in Merkel cell carcinoma. PMID- 18408657 TI - Array comparative genomic hybridization analysis of olfactory neuroblastoma. AB - Olfactory neuroblastoma is an unusual neuroectodermal malignancy, which is thought to arise at the olfactory membrane of the sinonasal tract. Due to its rarity, little is understood regarding its molecular and cytogenetic abnormalities. The aim of the current study is to identify specific DNA copy number changes in olfactory neuroblastoma. Thirteen dissected tissue samples were analyzed using array comparative genomic hybridization. Our results show that gene copy number profiles of olfactory neuroblastoma samples are complex. The most frequent changes included gains at 7q11.22-q21.11, 9p13.3, 13q, 20p/q, and Xp/q, and losses at 2q31.1, 2q33.3, 2q37.1, 6q16.3, 6q21.33, 6q22.1, 22q11.23, 22q12.1, and Xp/q. Gains were more frequent than losses, and high-stage tumors showed more alterations than low-stage olfactory neuroblastoma. Frequent changes in high-stage tumors were gains at 13q14.2-q14.3, 13q31.1, and 20q11.21-q11.23, and loss of Xp21.1 (in 66% of cases). Gains at 5q35, 13q, and 20q, and losses at 2q31.1, 2q33.3, and 6q16-q22, were present in 50% of cases. The identified regions of gene copy number change have been implicated in a variety of tumors, especially carcinomas. In addition, our results indicate that gains in 20q and 13q may be important in the progression of this cancer, and that these regions possibly harbor genes with functional relevance in olfactory neuroblastoma. PMID- 18408658 TI - Idiopathic pleuroparenchymal fibroelastosis: an unrecognized or misdiagnosed entity? AB - Idiopathic pleuroparenchymal fibroelastosis is a rare recently described entity likely to be under- and misdiagnosed, as awareness of this entity is not yet widespread. We report two cases that show the need to include this disease in the differential diagnosis of patients with predominantly pleural and subpleural fibrotic processes. The condition is a fibrotic thickening of the pleura and subpleural parenchyma due to elastic fiber proliferation predominantly in the upper lobes. Performing elastic fiber stains routinely in patients with fibrosis of this distribution may, therefore, aid in establishing the diagnosis and differentiating it from usual interstitial pneumonia/idiopathic pulmonary fibrosis. These patients may be prone to the development of secondary spontaneous pneumothoraces and persistent postoperative bronchopleural fistulae. Continued study of newly diagnosed cases may uncover shared characteristics or features helpful in generating an etiologic hypothesis. Only with better understanding of this disease can we hope in the future to be able to offer treatments other than supportive care and ultimately lung transplantation, which are the only therapeutic options available today. PMID- 18408659 TI - In melanocytic lesions the fraction of BRAF V600E alleles is associated with sun exposure but unrelated to ERK phosphorylation. AB - BRAF(V600E) mutation has been frequently reported in different types of melanocytic lesions, but its role in melanomagenesis is poorly understood, having been associated with either the proliferative-induced MAPK pathway activation or the acquisition of oncogene-driven senescence. The presence of BRAF alterations has been related to sun exposure, although the molecular mechanisms underlying this event are only partly known. To elucidate the relationships among BRAF/NRAS alterations, MAPK pathway activation, and sun exposure, we examined 22 acquired nevi and 18 cutaneus melanomas from 38 patients. Microdissected tissues from each lesion were subjected to BRAF/NRAS mutation analysis by sequencing, allele specific PCR and pyrosequencing assay. The same lesions were also examined for the expression of phosphorylated ERK1/2. Phototype and an accurate history of sun exposure were evaluated for each patient. BRAF(V600E) mutation was detected in 50% of the acquired nevi and in 70% of the cutaneus melanomas in the absence of NRAS alterations. The fraction of alleles carrying BRAF(V600E) substitution was variable but strongly associated with sun exposure. In contrast, no relationship was evidenced between the presence of this mutation and patients' phototype, phosphorylated ERK1/2 expression, or Clark's level. Our findings indicate that in melanocytic lesions, BRAF(V600E) mutation can affect a subset of the cells and is associated with the type and quantity of sun exposure. This mutation is independent of the nevo-melanoma progression and unrelated to ERK phosphorylation, suggesting that alternative mechanisms to the MAPK activation are also involved in this type of transformation. PMID- 18408660 TI - Tussock jumping... PMID- 18408661 TI - Breast is best. PMID- 18408662 TI - Communication failure. PMID- 18408663 TI - High speed laceration. PMID- 18408664 TI - Informed consent. PMID- 18408665 TI - Courageous decision. PMID- 18408666 TI - Micro-implants. PMID- 18408667 TI - Tongue lacerations. PMID- 18408668 TI - Pigeon-holing. PMID- 18408669 TI - Recipe for confusion. PMID- 18408670 TI - Aesthetic options. PMID- 18408671 TI - SAS contract. PMID- 18408681 TI - Cocaine and oral health. AB - In the UK almost one million individuals use cocaine on a regular basis, implying that dentists are likely to encounter individuals that use cocaine. Regular use of this drug may have several orofacial effects, such as perforation of the nasal septum and palate, gingival lesions and erosion of tooth surfaces. In addition, recent use of cocaine increases the risk of a medical emergency during dental treatment, especially when epinephrine-containing local anaesthetics or retraction cords are used. Therefore, dental treatment should be postponed for 6 to 24 hours after the use of cocaine. PMID- 18408682 TI - Conducting qualitative interviews with school children in dental research. AB - This paper explores when and how to conduct qualitative research with children in dentistry, using research interviews as a method of data collection. Methodological guidance is also provided, based on research and a recent dental public health study in which 74 children from Years 2 (6-7-year-olds) and 6 (10 11-year-olds) were interviewed about their understanding of and attitudes to food. PMID- 18408683 TI - Immediately loaded full-arch provisional implant restorations using CAD/CAM and guided placement: maxillary and mandibular case reports. AB - This paper describes a protocol which incorporates the benefits of medical imaging, computerised virtual treatment planning, computer guided surgery, and the fitting and immediate loading of a preformed provisional full arch fixed prosthesis. The protocol can be adapted for use with the implant system of the operator's choosing and a prosthetic carrier presents the immediate load prosthesis to the mouth in its correct three-dimensional inter-occlusal relationship, simplifying the prosthetic stages for immediate load case management. Two case studies are described to illustrate the procedure for both mandibular and maxillary reconstructions. PMID- 18408689 TI - Digital clinical records and practice administration in primary dental care. AB - Usually, a 'computerised dental practice' has included a series of diagnostic instruments, intra-oral cameras, digital radiographic systems, treatment planning systems, CAD-CAM systems, management systems etc. However, these 'island solutions' have not been integrated into one system. Nevertheless, it is possible to produce fully integrated systems for digital clinical records, based on established physiologic and cognitive-ergonomic concepts. The first part of this paper outlines the philosophy behind the development of such a totally integrated system for digital clinical records. The second--digital practice administration- considers how the 'digital revolution' has impacted upon practice administration. PMID- 18408704 TI - Aesthetic inlays and onlays: the coming of age. AB - There are many prominent teaching clinicians who feel that inlays and onlays (of whatever colour) are a grossly under-utilised restoration and that crowns are an over-utilised restoration. I think it is worthwhile to examine some of the possible reasons for this unfortunate situation (for our patients) and see if the reasons for dentists' reluctance to incorporate these restorations into their routine services are really valid today. PMID- 18408707 TI - An in vitro comparison of tooth whitening techniques on natural tooth colour. AB - OBJECTIVE: Tooth whitening has become a popular treatment regime but there is little quantitative evidence to compare techniques and so confusion may exist for the clinician as to which regime to prescribe for greatest efficacy. The aim of this study was to compare immediate and longer-term colour change on natural tooth colour in vitro, using five current tooth whitening techniques with blind matched control groups. METHODS: A total of 100 human teeth of matched size were cleaned, stored in sterile deionised water at 4 degrees C then randomly allocated to one of the five active treatment groups or five matched control groups. The active treatments were: 10% carbamide peroxide (CP) x 60 min, 35% CP x 30 min or 35% hydrogen peroxide (HP) treatment x 30 min activated by one of three sources of energy (diode laser, halogen light, and plasma arc curing light). Tooth colour was analysed with a colorimeter before and after treatment: immediate, one week and nine months post-bleaching designed to generate tooth colour value (L(*)) according to the L(*)a(*)b system. The change in colour was determined as DeltaL (the difference in the value of the colour) for each tooth, then the mean differences were obtained for each group and compared. Tooth surface temperature was monitored. RESULTS: Comparing active treatments with controls it was found that 10% CP, 35% CP, 35% HP with halogen provided significantly greater tooth whitening. Comparing the different treatments showed that 10% CP was significantly more effective (P <0.05) than all other treatments except 35% HP with halogen activation. The effect of each treatment regime over time showed that the 10% CP gave a significant gain immediately and one week later (P <0.05), however, all the whitening effects were lost over time following these single treatments. The temperature rise on the tooth surface was greatest when using laser activation during power whitening. CONCLUSION: This study suggests that 10% CP is an effective technique for tooth whitening and can offer significant benefits over alternative regimes. PMID- 18408708 TI - Nucleosome organization in the Drosophila genome. AB - Comparative genomics of nucleosome positions provides a powerful means for understanding how the organization of chromatin and the transcription machinery co-evolve. Here we produce a high-resolution reference map of H2A.Z and bulk nucleosome locations across the genome of the fly Drosophila melanogaster and compare it to that from the yeast Saccharomyces cerevisiae. Like Saccharomyces, Drosophila nucleosomes are organized around active transcription start sites in a canonical -1, nucleosome-free region, +1 arrangement. However, Drosophila does not incorporate H2A.Z into the -1 nucleosome and does not bury its transcriptional start site in the +1 nucleosome. At thousands of genes, RNA polymerase II engages the +1 nucleosome and pauses. How the transcription initiation machinery contends with the +1 nucleosome seems to be fundamentally different across major eukaryotic lines. PMID- 18408709 TI - Functional genomic screen reveals genes involved in lipid-droplet formation and utilization. AB - Eukaryotic cells store neutral lipids in cytoplasmic lipid droplets enclosed in a monolayer of phospholipids and associated proteins. These dynamic organelles serve as the principal reservoirs for storing cellular energy and for the building blocks for membrane lipids. Excessive lipid accumulation in cells is a central feature of obesity, diabetes and atherosclerosis, yet remarkably little is known about lipid-droplet cell biology. Here we show, by means of a genome wide RNA interference (RNAi) screen in Drosophila S2 cells that about 1.5% of all genes function in lipid-droplet formation and regulation. The phenotypes of the gene knockdowns sorted into five distinct phenotypic classes. Genes encoding enzymes of phospholipid biosynthesis proved to be determinants of lipid-droplet size and number, suggesting that the phospholipid composition of the monolayer profoundly affects droplet morphology and lipid utilization. A subset of the Arf1 COPI vesicular transport proteins also regulated droplet morphology and lipid utilization, thereby identifying a previously unrecognized function for this machinery. These phenotypes are conserved in mammalian cells, suggesting that insights from these studies are likely to be central to our understanding of human diseases involving excessive lipid storage. PMID- 18408710 TI - BCR-ABL1 lymphoblastic leukaemia is characterized by the deletion of Ikaros. AB - The Philadelphia chromosome, a chromosomal abnormality that encodes BCR-ABL1, is the defining lesion of chronic myelogenous leukaemia (CML) and a subset of acute lymphoblastic leukaemia (ALL). To define oncogenic lesions that cooperate with BCR-ABL1 to induce ALL, we performed a genome-wide analysis of diagnostic leukaemia samples from 304 individuals with ALL, including 43 BCR-ABL1 B progenitor ALLs and 23 CML cases. IKZF1 (encoding the transcription factor Ikaros) was deleted in 83.7% of BCR-ABL1 ALL, but not in chronic-phase CML. Deletion of IKZF1 was also identified as an acquired lesion at the time of transformation of CML to ALL (lymphoid blast crisis). The IKZF1 deletions resulted in haploinsufficiency, expression of a dominant-negative Ikaros isoform, or the complete loss of Ikaros expression. Sequencing of IKZF1 deletion breakpoints suggested that aberrant RAG-mediated recombination is responsible for the deletions. These findings suggest that genetic lesions resulting in the loss of Ikaros function are an important event in the development of BCR-ABL1 ALL. PMID- 18408711 TI - Drosophila odorant receptors are both ligand-gated and cyclic-nucleotide activated cation channels. AB - From worm to man, many odorant signals are perceived by the binding of volatile ligands to odorant receptors that belong to the G-protein-coupled receptor (GPCR) family. They couple to heterotrimeric G-proteins, most of which induce cAMP production. This second messenger then activates cyclic-nucleotide-gated ion channels to depolarize the olfactory receptor neuron, thus providing a signal for further neuronal processing. Recent findings, however, have challenged this concept of odorant signal transduction in insects, because their odorant receptors, which lack any sequence similarity to other GPCRs, are composed of conventional odorant receptors (for example, Or22a), dimerized with a ubiquitously expressed chaperone protein, such as Or83b in Drosophila. Or83b has a structure akin to GPCRs, but has an inverted orientation in the plasma membrane. However, G proteins are expressed in insect olfactory receptor neurons, and olfactory perception is modified by mutations affecting the cAMP transduction pathway. Here we show that application of odorants to mammalian cells co expressing Or22a and Or83b results in non-selective cation currents activated by means of an ionotropic and a metabotropic pathway, and a subsequent increase in the intracellular Ca(2+) concentration. Expression of Or83b alone leads to functional ion channels not directly responding to odorants, but being directly activated by intracellular cAMP or cGMP. Insect odorant receptors thus form ligand-gated channels as well as complexes of odorant-sensing units and cyclic nucleotide-activated non-selective cation channels. Thereby, they provide rapid and transient as well as sensitive and prolonged odorant signalling. PMID- 18408712 TI - Insect olfactory receptors are heteromeric ligand-gated ion channels. AB - In insects, each olfactory sensory neuron expresses between one and three ligand binding members of the olfactory receptor (OR) gene family, along with the highly conserved and broadly expressed Or83b co-receptor. The functional insect OR consists of a heteromeric complex of unknown stoichiometry but comprising at least one variable odorant-binding subunit and one constant Or83b family subunit. Insect ORs lack homology to G-protein-coupled chemosensory receptors in vertebrates and possess a distinct seven-transmembrane topology with the amino terminus located intracellularly. Here we provide evidence that heteromeric insect ORs comprise a new class of ligand-activated non-selective cation channels. Heterologous cells expressing silkmoth, fruitfly or mosquito heteromeric OR complexes showed extracellular Ca2+ influx and cation-non selective ion conductance on stimulation with odorant. Odour-evoked OR currents are independent of known G-protein-coupled second messenger pathways. The fast response kinetics and OR-subunit-dependent K+ ion selectivity of the insect OR complex support the hypothesis that the complex between OR and Or83b itself confers channel activity. Direct evidence for odorant-gated channels was obtained by outside-out patch-clamp recording of Xenopus oocyte and HEK293T cell membranes expressing insect OR complexes. The ligand-gated ion channel formed by an insect OR complex seems to be the basis for a unique strategy that insects have acquired to respond to the olfactory environment. PMID- 18408713 TI - Identification of RIP1 kinase as a specific cellular target of necrostatins. AB - Necroptosis is a cellular mechanism of necrotic cell death induced by apoptotic stimuli in the form of death domain receptor engagement by their respective ligands under conditions where apoptotic execution is prevented. Although it occurs under regulated conditions, necroptotic cell death is characterized by the same morphological features as unregulated necrotic death. Here we report that necrostatin-1, a previously identified small-molecule inhibitor of necroptosis, is a selective allosteric inhibitor of the death domain receptor-associated adaptor kinase RIP1 in vitro. We show that RIP1 is the primary cellular target responsible for the antinecroptosis activity of necrostatin-1. In addition, we show that two other necrostatins, necrostatin-3 and necrostatin-5, also target the RIP1 kinase step in the necroptosis pathway, but through mechanisms distinct from that of necrostatin-1. Overall, our data establish necrostatins as the first in-class inhibitors of RIP1 kinase, the key upstream kinase involved in the activation of necroptosis. PMID- 18408714 TI - Structural and biochemical evidence for a boat-like transition state in beta mannosidases. AB - Enzyme inhibition through mimicry of the transition state is a major area for the design of new therapeutic agents. Emerging evidence suggests that many retaining glycosidases that are active on alpha- or beta-mannosides harness unusual B2,5 (boat) transition states. Here we present the analysis of 25 putative beta mannosidase inhibitors, whose Ki values range from nanomolar to millimolar, on the Bacteroides thetaiotaomicron beta-mannosidase BtMan2A. B2,5 or closely related conformations were observed for all tightly binding compounds. Subsequent linear free energy relationships that correlate log Ki with log Km/kcat for a series of active center variants highlight aryl-substituted mannoimidazoles as powerful transition state mimics in which the binding energy of the aryl group enhances both binding and the degree of transition state mimicry. Support for a B2,5 transition state during enzymatic beta-mannosidase hydrolysis should also facilitate the design and exploitation of transition state mimics for the inhibition of retaining alpha-mannosidases--an area that is emerging for anticancer therapeutics. PMID- 18408715 TI - Unconscious determinants of free decisions in the human brain. AB - There has been a long controversy as to whether subjectively 'free' decisions are determined by brain activity ahead of time. We found that the outcome of a decision can be encoded in brain activity of prefrontal and parietal cortex up to 10 s before it enters awareness. This delay presumably reflects the operation of a network of high-level control areas that begin to prepare an upcoming decision long before it enters awareness. PMID- 18408716 TI - Reduction of stimulus visibility compresses apparent time intervals. AB - The neural mechanisms underlying visual estimation of subsecond durations remain unknown, but perisaccadic underestimation of interflash intervals may provide a clue as to the nature of these mechanisms. Here we found that simply reducing the flash visibility, particularly the visibility of transient signals, induced similar time underestimation by human observers. Our results suggest that weak transient responses fail to trigger the proper detection of temporal asynchrony, leading to increased perception of simultaneity and apparent time compression. PMID- 18408717 TI - Double dissociation of 'what' and 'where' processing in auditory cortex. AB - Studies of cortical connections or neuronal function in different cerebral areas support the hypothesis that parallel cortical processing streams, similar to those identified in visual cortex, may exist in the auditory system. However, this model has not yet been behaviorally tested. We used reversible cooling deactivation to investigate whether the individual regions in cat nonprimary auditory cortex that are responsible for processing the pattern of an acoustic stimulus or localizing a sound in space could be doubly dissociated in the same animal. We found that bilateral deactivation of the posterior auditory field resulted in deficits in a sound-localization task, whereas bilateral deactivation of the anterior auditory field resulted in deficits in a pattern-discrimination task, but not vice versa. These findings support a model of cortical organization that proposes that identifying an acoustic stimulus ('what') and its spatial location ('where') are processed in separate streams in auditory cortex. PMID- 18408718 TI - Mutations in TMPRSS6 cause iron-refractory iron deficiency anemia (IRIDA). AB - Iron deficiency is usually attributed to chronic blood loss or inadequate dietary intake. Here, we show that iron deficiency anemia refractory to oral iron therapy can be caused by germline mutations in TMPRSS6, which encodes a type II transmembrane serine protease produced by the liver that regulates the expression of the systemic iron regulatory hormone hepcidin. These findings demonstrate that TMPRSS6 is essential for normal systemic iron homeostasis in humans. PMID- 18408719 TI - Exposing the fitness contribution of duplicated genes. AB - Duplicate genes from the whole-genome duplication (WGD) in yeast are often dispensable--removing one copy has little or no phenotypic consequence. It is unknown, however, whether such dispensability reflects insignificance of the ancestral function or compensation from paralogs. Here, using precise competition based measurements of the fitness cost of single and double deletions, we estimate the exposed fitness contribution of WGD duplicate genes in metabolism and bound the importance of their ancestral pre-duplication function. We find that the functional overlap between paralogs sufficiently explains the apparent dispensability of individual WGD genes. Furthermore, the lower bound on the fitness value of the ancestral function, which is estimated by the degree of synergistic epistasis, is at least as large as the average fitness cost of deleting single non-WGD genes. These results suggest that most metabolic functions encoded by WGD genes are important today and were also important at the time of duplication. PMID- 18408720 TI - No evidence of clonal somatic genetic alterations in cancer-associated fibroblasts from human breast and ovarian carcinomas. AB - There is increasing evidence showing that the stromal cells surrounding cancer epithelial cells, rather than being passive bystanders, might have a role in modifying tumor outgrowth. The molecular basis of this aspect of carcinoma etiology is controversial. Some studies have reported a high frequency of genetic aberrations in carcinoma-associated fibroblasts (CAFs), whereas other studies have reported very low or zero mutation rates. Resolution of this contentious area is of critical importance in terms of understanding both the basic biology of cancer as well as the potential clinical implications of CAF somatic alterations. We undertook genome-wide copy number and loss of heterozygosity (LOH) analysis of CAFs derived from breast and ovarian carcinomas using a 500K SNP array platform. Our data show conclusively that LOH and copy number alterations are extremely rare in CAFs and cannot be the basis of the carcinoma promoting phenotypes of breast and ovarian CAFs. PMID- 18408721 TI - Skint1, the prototype of a newly identified immunoglobulin superfamily gene cluster, positively selects epidermal gammadelta T cells. AB - B cells, alphabeta T cells and gammadelta T cells are conserved lymphocyte subtypes encoding their antigen receptors from somatically rearranged genes. alphabeta T cells undergo positive selection in the thymus by engagement of their T cell receptors (TCRs) with self-peptides presented by major histocompatibility complex molecules. The molecules that select gammadelta T cells are unknown. Vgamma5+Vdelta1+ cells comprise 90% of mouse epidermal gammadelta T cells. By mapping and genetic complementation using a strain showing loss of Vgamma5+Vdelta1+ cells due to a failure of thymic selection, we show that this defect is caused by mutation in Skint1, a newly identified gene expressed in thymus and skin that encodes a protein with immunoglobulin-like and transmembrane domains. Skint1 is the prototypic member of a rapidly evolving family of at least 11 genes in mouse, with greatest similarity to the butyrophilin genes. These findings define a new family of proteins mediating key epithelial-immune interactions. PMID- 18408722 TI - The proline-rich sequence of CD3epsilon controls T cell antigen receptor expression on and signaling potency in preselection CD4+CD8+ thymocytes. AB - Antigen recognition by T cell antigen receptors (TCRs) is thought to 'unmask' a proline-rich sequence (PRS) present in the CD3epsilon cytosolic segment, which allows it to trigger T cell activation. Using 'knock-in' mice with deletion of the PRS, we demonstrate here that elimination of the CD3epsilon PRS had no effect on mature T cell responsiveness. In contrast, in preselection CD4+CD8+ thymocytes, the CD3epsilon PRS acted together with the adaptor protein SLAP to promote CD3zeta degradation, thereby contributing to downregulation of TCR expression on the cell surface. In addition, analysis of CD4+CD8+ thymocytes of TCR-transgenic mice showed that the CD3epsilon PRS enhanced TCR sensitivity to weak ligands. Our results identify previously unknown functions for the evolutionarily conserved CD3epsilon PRS at the CD4+CD8+ developmental stage and suggest a rather limited function in mature T cells. PMID- 18408723 TI - A zeolite family with chiral and achiral structures built from the same building layer. AB - Porosity and chirality are two of the most important properties for materials in the chemical and pharmaceutical industry. Inorganic microporous materials such as zeolites have been widely used in ion-exchange, selective sorption/separation and catalytic processes. The pore size and shape in zeolites play important roles for specific applications. Chiral inorganic microporous materials are particularly desirable with respect to their possible use in enantioselective sorption, separation and catalysis. At present, among the 179 zeolite framework types reported, only three exhibit chiral frameworks. Synthesizing enantiopure, porous tetrahedral framework structures represents a great challenge for chemists. Here, we report the silicogermanates SU-32 (polymorph A), SU-15 (polymorph B) (SU, Stockholm University) and a hypothetical polymorph C, all built by different stacking of a novel building layer. Whereas polymorphs B and C are achiral, each crystal of polymorph A exhibits only one hand and has an intrinsically chiral zeolite structure. SU-15 and SU-32 are thermally stable on calcination. PMID- 18408724 TI - Simultaneous optimization of charge-carrier mobility and optical gain in semiconducting polymer films. AB - The combination of efficient light emission and high charge-carrier mobility has thus far proved elusive for polymer semiconductors, with high mobility typically achieved by cofacial pi-electron system to pi-electron system interactions that quench exciton luminescence. We report a new strategy, comprising the introduction of a limited number of more effective hopping sites between otherwise relatively isolated, and thus highly luminescent, polyfluorene chains. Our approach results in polymer films with large mobility (mu approximately 3-6 x 10(-2) cm2 V-1 s-1) and simultaneously excellent light-emission characteristics. These materials are expected to be of interest for light-emitting transistors, light-emitting diode sources for optical communications and may offer renewed hope for electrically pumped laser action. In the last context, optically pumped distributed feedback lasers comprising one-dimensional etched silica grating structures coated with polymer have state-of-the-art excitation thresholds (as low as 30 W cm(-2) (0.1 nJ per pulse or 0.3 microJ cm-2) for 10 Hz, 12 ns, 390 nm excitation) and slope efficiencies (up to 11%). PMID- 18408725 TI - Femtosecond laser nanoaxotomy lab-on-a-chip for in vivo nerve regeneration studies. AB - A thorough understanding of nerve regeneration in Caenorhabditis elegans requires performing femtosecond laser nanoaxotomy while minimally affecting the worm. We present a microfluidic device that fulfills such criteria and can easily be automated to enable high-throughput genetic and pharmacological screenings. Using the 'nanoaxotomy' chip, we discovered that axonal regeneration occurs much faster than previously described, and notably, the distal fragment of the severed axon regrows in the absence of anesthetics. PMID- 18408726 TI - Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics. AB - We demonstrate live-cell super-resolution imaging using photoactivated localization microscopy (PALM). The use of photon-tolerant cell lines in combination with the high resolution and molecular sensitivity of PALM permitted us to investigate the nanoscale dynamics within individual adhesion complexes (ACs) in living cells under physiological conditions for as long as 25 min, with half of the time spent collecting the PALM images at spatial resolutions down to approximately 60 nm and frame rates as short as 25 s. We visualized the formation of ACs and measured the fractional gain and loss of individual paxillin molecules as each AC evolved. By allowing observation of a wide variety of nanoscale dynamics, live-cell PALM provides insights into molecular assembly during the initiation, maturation and dissolution of cellular processes. PMID- 18408727 TI - Suspended-drop electroporation for high-throughput delivery of biomolecules into cells. AB - We present a high-throughput method that enables efficient delivery of biomolecules into cells. The device consists of an array of 96 suspended electrode pairs, where small sample volumes are top-loaded, electroporated and bottom-ejected into 96-well plates. We demonstrate the use of this suspended-drop electroporation (SDE) device to effectively introduce fluorescent dextran, small interfering RNA (siRNA) or cDNA into primary neurons, differentiated neutrophils and other cell types with conventionally low transfection rates. PMID- 18408729 TI - Replacing PCR with COLD-PCR enriches variant DNA sequences and redefines the sensitivity of genetic testing. AB - PCR is widely employed as the initial DNA amplification step for genetic testing. However, a key limitation of PCR-based methods is the inability to selectively amplify low levels of mutations in a wild-type background. As a result, downstream assays are limited in their ability to identify subtle genetic changes that can have a profound impact in clinical decision-making and outcome. Here we describe co-amplification at lower denaturation temperature PCR (COLD-PCR), a novel form of PCR that amplifies minority alleles selectively from mixtures of wild-type and mutation-containing sequences irrespective of the mutation type or position on the sequence. We replaced regular PCR with COLD-PCR before sequencing or genotyping assays to improve mutation detection sensitivity by up to 100-fold and identified new mutations in the genes encoding p53, KRAS and epidermal growth factor in heterogeneous cancer samples that had been missed by the currently used methods. For clinically relevant microdeletions, COLD-PCR enabled exclusive amplification and isolation of the mutants. COLD-PCR will transform the capabilities of PCR-based genetic testing, including applications in cancer, infectious diseases and prenatal identification of fetal alleles in maternal blood. PMID- 18408730 TI - ATP-dependent chromatin remodeling shapes the DNA replication landscape. AB - The eukaryotic DNA replication machinery must traverse every nucleosome in the genome during S phase. As nucleosomes are generally inhibitory to DNA-dependent processes, chromatin structure must undergo extensive reorganization to facilitate DNA synthesis. However, the identity of chromatin-remodeling factors involved in replication and how they affect DNA synthesis is largely unknown. Here we show that two highly conserved ATP-dependent chromatin-remodeling complexes in Saccharomyces cerevisiae, Isw2 and Ino80, function in parallel to promote replication fork progression. As a result, Isw2 and Ino80 have especially important roles for replication of late-replicating regions during periods of replication stress. Both Isw2 and Ino80 complexes are enriched at sites of replication, suggesting that these complexes act directly to promote fork progression. These findings identify ATP-dependent chromatin-remodeling complexes that promote DNA replication and define a specific stage of replication that requires remodeling for normal function. PMID- 18408731 TI - DNA apurinic-apyrimidinic site binding and excision by endonuclease IV. AB - Escherichia coli endonuclease IV is an archetype for an abasic or apurinic apyrimidinic endonuclease superfamily crucial for DNA base excision repair. Here biochemical, mutational and crystallographic characterizations reveal a three metal ion mechanism for damage binding and incision. The 1.10-A resolution DNA free and the 2.45-A resolution DNA-substrate complex structures capture substrate stabilization by Arg37 and reveal a distorted Zn3-ligand arrangement that reverts, after catalysis, to an ideal geometry suitable to hold rather than release cleaved DNA product. The 1.45-A resolution DNA-product complex structure shows how Tyr72 caps the active site, tunes its dielectric environment and promotes catalysis by Glu261-activated hydroxide, bound to two Zn2+ ions throughout catalysis. These structural, mutagenesis and biochemical results suggest general requirements for abasic site removal in contrast to features specific to the distinct endonuclease IV alpha-beta triose phosphate isomerase (TIM) barrel and APE1 four-layer alpha-beta folds of the apurinic-apyrimidinic endonuclease families. PMID- 18408732 TI - The HSA domain binds nuclear actin-related proteins to regulate chromatin remodeling ATPases. AB - We identify the helicase-SANT-associated (HSA) domain as the primary binding platform for nuclear actin-related proteins (ARPs) and actin. Individual HSA domains from chromatin remodelers (RSC, yeast SWI-SNF, human SWI-SNF, SWR1 and INO80) or modifiers (NuA4) reconstitute their respective ARP-ARP or ARP-actin modules. In RSC, the HSA domain resides on the catalytic ATPase subunit Sth1. The Sth1 HSA is essential in vivo, and its omission causes the specific loss of ARPs and a moderate reduction in ATPase activity. Genetic selections for arp suppressors yielded specific gain-of-function mutations in two new domains in Sth1, the post-HSA domain and protrusion 1, which are essential for RSC function in vivo but not ARP association. Taken together, we define the role of the HSA domain and provide evidence for a regulatory relationship involving the ARP-HSA module and two new functional domains conserved in remodeler ATPases that contain ARPs. PMID- 18408733 TI - Arsenic degrades PML or PML-RARalpha through a SUMO-triggered RNF4/ubiquitin mediated pathway. AB - In acute promyelocytic leukaemia (APL), arsenic trioxide induces degradation of the fusion protein encoded by the PML-RARA oncogene, differentiation of leukaemic cells and produces clinical remissions. SUMOylation of its PML moiety was previously implicated, but the nature of the degradation pathway involved and the role of PML-RARalpha catabolism in the response to therapy have both remained elusive. Here, we demonstrate that arsenic-induced PML SUMOylation triggers its Lys 48-linked polyubiquitination and proteasome-dependent degradation. When exposed to arsenic, SUMOylated PML recruits RNF4, the human orthologue of the yeast SUMO-dependent E3 ubiquitin-ligase, as well as ubiquitin and proteasomes onto PML nuclear bodies. Arsenic-induced differentiation is impaired in cells transformed by a non-degradable PML-RARalpha SUMOylation mutant or in APL cells transduced with a dominant-negative RNF4, directly implicating PML-RARalpha catabolism in the therapeutic response. We thus identify PML as the first protein degraded by SUMO-dependent polyubiquitination. As PML SUMOylation recruits not only RNF4, ubiquitin and proteasomes, but also many SUMOylated proteins onto PML nuclear bodies, these domains could physically integrate the SUMOylation, ubiquitination and degradation pathways. PMID- 18408735 TI - TH17 cells in development: an updated view of their molecular identity and genetic programming. AB - Following activation, CD4+ T cells differentiate into different lineages of helper T (T(H)) cells that are characterized by distinct developmental regulation and biological functions. T(H)17 cells have recently been identified as a new lineage of effector T(H) cells, and they have been shown to be important in immune responses to infectious agents, as well as in various immune diseases. Over the past two to three years, there has been a rapid progress in our understanding of the differentiation programme of T(H)17 cells. Here, I summarize our current knowledge of the unique gene expression, cytokine-mediated regulation and transcriptional programming of T(H)17 cells, and provide my personal perspectives on the future studies that are required to elucidate this lineage in more detail. PMID- 18408734 TI - RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation. AB - In acute promyelocytic leukaemia (APL), the promyelocytic leukaemia (PML) protein is fused to the retinoic acid receptor alpha (RAR). This disease can be treated effectively with arsenic, which induces PML modification by small ubiquitin-like modifiers (SUMO) and proteasomal degradation. Here we demonstrate that the RING domain-containing ubiquitin E3 ligase, RNF4 (also known as SNURF), targets poly SUMO-modified proteins for degradation mediated by ubiquitin. RNF4 depletion or proteasome inhibition led to accumulation of mixed, polyubiquitinated, poly-SUMO chains. PML protein accumulated in RNF4-depleted cells and was ubiquitinated by RNF4 in a SUMO-dependent fashion in vitro. In the absence of RNF4, arsenic failed to induce degradation of PML and SUMO-modified PML accumulated in the nucleus. These results demonstrate that poly-SUMO chains can act as discrete signals from mono-SUMOylation, in this case targeting a poly-SUMOylated substrate for ubiquitin-mediated proteolysis. PMID- 18408736 TI - Association between weight perception and psychological distress. AB - BACKGROUND: Obesity is a well-known cause of cardiovascular disease burden and premature death, but effects on depressive symptoms remain equivocal. Depressive symptoms may be more common among the obese individuals who perceive themselves as overweight, rather than those who perceive themselves as having an acceptable weight. Our aim was to determine whether weight status and weight perceptions are independently associated with psychological distress. METHODS: We conducted a cross-sectional study using data from the Australian National Health Survey 2004 2005 (N=17 253). All variables were collected by self-report. Adjusted multinomial logistic regression analysis was conducted to generate prevalence odds ratios with 95% confidence intervals (95% CI) for medium (Kessler Psychological Distress Scale (K10) scores of 20-29) and high (K10 scores of 30 50) psychological distress (compared with K10 scores of 10-19 as the reference) associated with weight status (standard body mass index (BMI) cutoffs for underweight, overweight and obesity vs normal weight), weight perception (perceived underweight and overweight vs acceptable weight) and weight misperception (incorrect with BMI vs correct with BMI) adjusting for numerous important covariates. RESULTS: Overweight and underweight perception increased the odds of medium (40 and 50%) and high (50 and 120%) psychological distress, whereas weight status and weight misperception were not associated with psychological distress in adjusted analysis. Gender, alcohol consumed per week and post-school education were not significant covariates (at P<0.10 level). CONCLUSIONS: Overweight and underweight perception rather than weight status or weight misperception are significant risk factors associated with medium and high psychological distress prevalence and effects appear to be uniform for men and women. Well-designed prospective studies are still needed to determine whether weight perceptions cause psychological distress, and if so, whether symptoms are significantly reduced following effective intervention. PMID- 18408737 TI - ABT-737, proving to be a great tool even before it is proven in the clinic. PMID- 18408742 TI - Wound healing without hair. PMID- 18408738 TI - p27 Kip1 nuclear localization and cyclin-dependent kinase inhibitory activity are regulated by glycogen synthase kinase-3 in human colon cancer cells. AB - The cellular mechanisms regulating intestinal differentiation are poorly understood. Sodium butyrate (NaBT), a short-chain fatty acid, increases p27 Kip1 expression and induces cell cycle arrest associated with intestinal cell differentiation. Here, we show that treatment of intestinal-derived cells with NaBT induced G0/G1 arrest and intestinal alkaline phosphatase, a marker of differentiation, activity and mRNA expression; this induction was attenuated by inhibition of glycogen synthase kinase-3 (GSK-3). Moreover, treatment with NaBT increased the nuclear, but not the cytosolic, expression and activity of GSK 3beta. NaBT decreased cyclin-dependent kinase CDK2 activity and induced p27 Kip1 expression; inhibition of GSK-3 rescued NaBT-inhibited CDK2 activity and blocked NaBT-induced p27 Kip1 expression in the nucleus but not in the cytoplasm. In addition, we demonstrate that NaBT decreased the expression of S-phase kinase associated protein 2 (Skp2), and this decrease was attenuated by GSK-3 inhibition. Furthermore, NaBT increased p27 Kip1 binding to CDK2, which was completely abolished by GSK-3 inhibition. Overexpression of an active form of GSK 3beta reduced Skp2 expression, increased p27 Kip1 in the nucleus and increased p27 Kip1 binding to CDK2. Our results suggest that GSK-3 not only regulates nuclear p27 Kip1 expression through the downregulation of nuclear Skp2 expression but also functions to regulate p27 Kip1 assembly with CDK2, thereby playing a critical role in the G0/G1 arrest associated with intestinal cell differentiation. PMID- 18408743 TI - Is the hair follicle necessary for normal wound healing? AB - The hair follicle contributes cells to the interfollicular epidermis after wounding, but the functional role of these cells has not been resolved. To address this question, Langton et al. (this issue, 2008) take advantage of the Edaradd mutant mouse, which lacks hair follicles on its tail. They discover an initial sluggish response of the hairless tail epidermis to wounding that is rapidly compensated for by recruitment of epidermal cells from outside the normally responsive area. This suggests that the hair follicle is important but not necessary for normal wound healing. PMID- 18408744 TI - Modeling atopic dermatitis with increasingly complex mouse models. AB - Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disorder that affects approximately 15% of children in the United States. A complex disorder, AD is characterized by both skin barrier impairment and immunologic abnormalities, including decreased innate immune function and a polarized adaptive immune response. Mouse models have demonstrated the complex interdependence of immune cell-keratinocyte interactions and teased apart gene environment relationships in a controlled setting. In this issue, Nagelkerken et al. present a mouse model with transgenic expression of apolipoprotein C1 that disrupts the skin lipid barrier and manifests many hallmark features of AD. PMID- 18408745 TI - T-helper 17 cells in psoriatic plaques and additional genetic links between IL-23 and psoriasis. AB - T-helper 17 (Th17) cells are a newly appreciated T-cell subset, distinct from both Th1 and Th2 cells, that have been implicated in the pathogenesis of psoriasis and other autoimmune inflammatory diseases (Figure 1) (Fitch et al., 2007; Kastelein et al., 2007). IL-23 stimulates survival and proliferation of Th17 cells and thus serves as a key master cytokine regulator for these diseases. PMID- 18408746 TI - "Outside-to-inside" (and now back to "outside") pathogenic mechanisms in atopic dermatitis. AB - The pathogenesis of atopic dermatitis (AD) has been attributed largely to abnormalities in the adaptive immune system, with key roles played by T-helper 1(Th1)/Th2 cell dysregulation, IgE production, dendritic cell signaling, and mast cell hyperactivity, resulting in the pruritic, inflammatory dermatosis that characterizes AD (Leung et al., 2004). Accordingly, therapy has been focused on ameliorating Th2-mediated inflammation and pruritus (eg, Leung, 2000). Indeed, there is emerging evidence that inflammation in AD results first from inherited and acquired insults that converge to alter epidermal structure and function, followed by immune system activation, which in turn has negative consequences for skin-barrier homeostasis. This cycle comprises an "outside-inside-outside" model of AD pathogenesis (Elias et al., in press). PMID- 18408748 TI - Epidemiologic support for melanoma heterogeneity using the surveillance, epidemiology, and end results program. PMID- 18408747 TI - Controlling hair follicle signaling pathways through polyubiquitination. AB - Hair follicle development and maintenance require precise reciprocal signaling interactions between the epithelium and underlying dermis. Three major developmental signaling pathways, Wnt, Sonic hedgehog, and NF-kappaB/Edar, are indispensable for this process and, when aberrantly activated, can lead to skin and appendage neoplasms. Recent data point to protein polyubiquitination as playing a central role in regulating the timing, duration, and location of signaling. Here we review how polyubiquitination regulates the stability and interaction of key signaling components that control hair follicle development and regeneration. PMID- 18408749 TI - Array CGH demonstrates characteristic aberration signatures in human papillary thyroid carcinomas governed by RET/PTC. AB - The aim of this study is to investigate additional genetic alterations in papillary thyroid carcinomas (PTCs) with known RET/PTC rearrangements. We applied array-based comparative genomic hybridization (array CGH) to 33 PTC (20 PTC from adults, 13 post-Chernobyl PTC from children) with known RET/PTC status. Principal component analysis and hierarchical cluster analysis identified cases with similar aberration patterns. Significant deviations between tumour-groups were obtained by statistical testing (Fisher's exact test in combination with Benjamini-Hochberg FDR-controlling procedure). FISH analysis on FFPE sections was applied to validate the array CGH data. Deletions were found more frequently in RET/PTC-positive and RET/PTC-negative tumours than amplifications. Specific aberration signatures were identified that discriminated between RET/PTC-positive and RET/PTC-negative cases (aberrations on chromosomes 1p, 3q, 4p, 7p, 9p/q, 10q, 12q, 13q and 21q). In addition, childhood and adult RET/PTC-positive cases differ significantly for a deletion on the distal part of chromosome 1p. There are additional alterations in RET/PTC-positive tumours, which may act as modifiers of RET activation. In contrast, alterations in RET/PTC-negative tumours indicate alternative routes of tumour development. The data presented serve as a starting point for further studies on gene expression and function of genes identified in this study. PMID- 18408750 TI - Mechanistic insight into taxol-induced cell death. AB - We analysed the involvement of proteases during taxol-mediated cell death of human A549 non-small-cell lung carcinoma cells using a proteomics approach that specifically targets protein N termini and further detects newly formed N termini that are the result of protein processing. Our analysis revealed 27 protease mediated cleavages, which we divided in sites C-terminal to aspartic acid (Asp) and sites C-terminal to non-Asp residues, as the result of caspase and non caspase protease activities, respectively. Remarkably, some of the former were insensitive to potent pancaspase inhibitors, and we therefore suggest that previous inhibitor-based studies that report on the caspase-independent nature of taxol-induced cell death should be judged with care. Furthermore, many of the sites C-terminal to non-Asp residues were also uniquely observed in a model of cytotoxic granule-mediated cell death and/or found by in vitro cataloging human mu-calpain substrates using a similar proteomics technique. This thus raises the hypothesis that killing tumor cells by chemotherapy or by immune cells holds similar non-Asp-specific proteolytic components with strong indications to calpain activity. PMID- 18408751 TI - The BH3-only member Noxa causes apoptosis in melanoma cells by multiple pathways. AB - The molecular causes for resistance of melanoma to apoptosis are currently only partly understood. In the present study, we examined gene transfer and expression of the proapoptotic BH3-only protein Noxa as an alternative approach to chemotherapy and investigated the molecular mechanisms regulating Noxa-induced apoptosis. Noxa gene transfer caused dysregulation of both mitochondria and, as shown for the first time, also the endoplasmic reticulum, resulting in the accumulation of reactive oxygen species. Interestingly, expression of Noxa not only triggered the classical mitochondrial caspase cascade, but also resulted in the activation of apoptosis signal-regulating kinase1 and its downstream effectors c-Jun N-terminal kinase and p38. The activation of these kinases was abolished by antioxidants. Moreover, inhibition of the kinases by RNA interference or pharmacological inhibitors significantly attenuated Noxa-induced apoptosis. Thus, our data provide evidence for the involvement of multiple pathways in Noxa-induced apoptosis that are triggered at mitochondria and the endoplasmic reticulum, and suggest Noxa gene transfer as a complementary approach to chemotherapy. PMID- 18408752 TI - DICKKOPF-4 is induced by TCF/beta-catenin and upregulated in human colon cancer, promotes tumour cell invasion and angiogenesis and is repressed by 1alpha,25 dihydroxyvitamin D3. AB - Aberrant activation of the Wnt/beta-catenin signaling pathway is a hallmark of colon cancer. We show that the Wnt antagonist DICKKOPF-4 (DKK-4) gene is repressed by 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) in human colon cancer cells. This effect correlated with the inhibition of the DKK-4 promoter. Chromatin immunoprecipitation assays revealed that 1,25(OH)2D3 induces early and transient binding of the vitamin D receptor (VDR) and the SMRT corepressor to the region adjacent to the transcription start site of DKK-4. Additionally, we demonstrate that the DKK-4 gene is a new downstream target of TCF/beta-catenin. Remarkably, expression of DKK-4 messenger RNA (mRNA) was not detected in a series of 29 human normal (N) colon biopsies but expression was upregulated in all the matched tumour (T) tissues. An inverse correlation existed between the expression of DKK-4 and VDR RNA in the Ts. Ectopic DKK-4 expression increased the migration and invasion properties of colon cancer cells and conditioned media (CM) from DKK 4-expressing cells enhanced the capacity to migrate and form capillary-like tubules of human primary microvascular endothelial cells. In conclusion, DKK-4 is upregulated in colon cancer and is associated with the acquisition of malignant properties by neoplastic cells. DKK-4 downregulation is a novel effect of 1,25(OH)2D3 that may contribute to its anticancer action. PMID- 18408753 TI - The transcription-repression domain of the adenovirus E1A oncoprotein targets p300 at the promoter. AB - Extensive mutational/functional analysis of the transcription-repression domain encoded in the N-terminal 80 amino acids of the adenovirus E1A 243R oncoprotein suggests a model for the molecular mechanism of E1A repression: E1A accesses transcriptional co-activators such as p300 on specific promoters and then interacts with TBP to disrupt the TBP-TATA complex. In support of this model, as reported here, a basal core promoter activated by tethering p300 is repressible by E1A at the promoter level as shown by chromatin immunoprecipitation (ChIP) analysis. Sequestration of p300 by E1A does not play a significant role, as indicated by dose-response measurements. Furthermore, when the core promoter is transcriptionally activated by tethering activation domains of several transcription factors that can recruit p300 (p65, MyoD, cMyb and TFE3), transcription is repressible by E1A. However, when the core promoter is activated by factors not known to recruit p300 (USF1 and USF2), transcription is resistant to E1A repression. Finally, tethering p300 to the non-repressible adenovirus major late promoter (MLP) renders it repressible by E1A. ChIP analysis shows that E1A occupies the repressed MLP. These findings provide support for the hypothesis that p300 can serve as a scaffold for the E1A repression domain to access specific cellular gene promoters involved in growth regulation. PMID- 18408754 TI - Histone H4 lysine 20 monomethylation promotes transcriptional repression by L3MBTL1. AB - Lethal 3 malignant brain tumor 1 (L3MBTL1), a homolog of the Drosophila polycomb tumor suppressor l(3)mbt, contains three tandem MBT repeats (3xMBT) that are critical for transcriptional repression. We recently reported that the 3xMBT repeats interact with mono- and dimethylated lysines in the amino termini of histones H4 and H1b to promote methylation-dependent chromatin compaction. Using a series of histone peptides, we now show that the recognition of mono- and dimethylated lysines in histones H3, H4 and H1.4 (but not their trimethylated or unmodified counterparts) by 3xMBT occurs in the context of a basic environment, requiring a conserved aspartic acid (D355) in the second MBT repeat. Despite the broad range of in vitro binding, the chromatin association of L3MBTL1 mirrors the progressive accumulation of H4K20 monomethylation during the cell cycle. Furthermore, transcriptional repression by L3MBTL1 is enhanced by the H4K20 monomethyltransferase PR-SET7 (to which it binds) but not SUV420H1 (an H4K20 trimethylase) or G9a (an H3K9 dimethylase) and knockdown of PR-SET7 decreases H4K20me1 levels and the chromatin association of L3MBTL1. Our studies identify the importance of H4K20 monomethylation and of PR-SET7 for L3MBTL1 function. PMID- 18408755 TI - Snai1 and Snai2 collaborate on tumor growth and metastasis properties of mouse skin carcinoma cell lines. AB - Snai1 (Snail) and Snai2 (Slug), the two main members of Snail family factors, are important mediators of epithelial-mesenchymal transitions and involved in tumor progression. We recently reported that Snai1 plays a major role in tumor growth, invasion and metastasis, but the contribution of Snai2 to tumorigenesis is not yet well understood. To approach this question we have silenced Snai2 and/or Snai1 by stable RNA interference in two independent mouse skin carcinoma (HaCa4 and CarB) cell lines. We demonstrate that Snai2 knockdown has a milder effect, but collaborates with Snai1 silencing in reduction of tumor growth potential of either carcinoma cell line when injected into nude mice. Importantly, Snai1 or Snai2 silencing dramatically influences the metastatic ability of squamous carcinoma HaCa4 cells, inducing a strong reduction in liver and lung distant metastasis. However, only Snai1 knockdown has an effective action on invasiveness and fully abolishes tumor cell dissemination into the spleen. These results demonstrate that Snai1 and Snai2 collaborate on primary tumor growth and specifically contribute to site-specific metastasis of HaCa4 cells. These data also indicate that Snai1 is the major regulator of local invasion, supporting a hierarchical participation of both factors in the metastatic process. PMID- 18408756 TI - E2F1 regulates autophagy and the transcription of autophagy genes. AB - The retinoblastoma pathway is often inactivated in human tumors resulting in deregulated E2F activity that can induce both proliferation and cell death. Although the role of E2F in apoptosis is well characterized, little is known regarding its putative participation in other cell death pathways. We show here that activation of E2F1 upregulates the expression of four autophagy genes microtubule-associated protein-1 light chain-3 (LC3), autophagy-related gene-1 (ATG1), ATG5 and damage-regulated autophagy modulator (DRAM). E2F1-mediated induction of LC3, ATG1 and DRAM is direct and indeed, endogenous E2F1 can be found bound to regions encompassing the promoters of these genes. Regulation of ATG5 by E2F1 is indirect. Importantly, we demonstrate that E2F1 activation enhances autophagy and conversely, reducing endogenous E2F1 expression inhibits DNA damage-induced autophagy. These studies identify E2F1 as a transcriptional regulator of autophagy, and for the first time establish a role for E2F1 in DNA damage-induced autophagy. PMID- 18408757 TI - Unbalanced translocation, a major chromosome alteration causing loss of heterozygosity in human lung cancer. AB - Loss of heterozygosity (LOH) is a major genetic event causing inactivation of tumor suppressor genes in human carcinogenesis. To elucidate chromosomal mechanisms causing LOH, 201 LOHs in 10 cases of human lung cancer, which were detected by a genome-wide single nucleotide polymorphism array analysis, were investigated for responsible chromosome alterations by integrating information on breakpoints for DNA copy number changes obtained by array-comparative genome hybridization and on numerical and structural chromosomal alterations obtained by spectral karyotyping. The majority (80%) of LOHs were partial chromosome LOHs caused by structural chromosomal alterations, while the remaining (20%) were whole chromosome LOHs caused by whole chromosome deletions. Unbalanced translocation was defined as the most frequent alteration, and it accounted for 30% of all LOHs. Three other structural alterations-interstitial deletion (19%), mitotic recombination (9%) and gene conversion (6%)-also contributed to the occurrence of LOH, while terminal deletion contributed to only a small subset (1%). Since unbalanced translocation is a common chromosomal alteration in lung cancer cells, the results in the present study strongly indicate that a considerable fraction of LOHs detected in lung cancer cells are caused by unbalanced translocation. PMID- 18408758 TI - Regulation of IKKbeta by miR-199a affects NF-kappaB activity in ovarian cancer cells. AB - Cancer progression is an abnormal form of tissue repair characterized by chronic inflammation. IkappaB kinase-beta (IKKbeta) required for nuclear factor-kappaB (NF-kappaB) activation plays a critical role in this process. Using EOC cells isolated from malignant ovarian cancer ascites and solid tumors, we identified IKKbeta as a major factor promoting a functional TLR-MyD88-NF-kappaB pathway that confers to EOC cell the capacity to constitutively secrete proinflammatory/protumor cytokines and therefore promoting tumor progression and chemoresistance. Furthermore, we describe for the first time the identification of the microRNA hsa-miR-199a as a regulator of IKKbeta expression. Our study describes the property of ovarian cancer cells to enhance the inflammatory microenvironment as a result of the expression of an active IKKbeta pathway. Identification of these markers in patients' tumor samples may facilitate the adequate selection of treatment and open new venues for the development of effective therapy for chemoresistant ovarian cancers. PMID- 18408759 TI - AID and RAG1 do not contribute to lymphomagenesis in Emu c-myc transgenic mice. AB - DNA breaks caused by recombination-activating gene 1 (RAG1) and activation induced cytidine deaminase (AID) induce c-myc/immunoglobulin (Ig) heavy chain chromosomal translocations and thereby stimulate lymphomagenesis. However, constitutive expression of c-myc alone is not sufficient to induce lymphomas. Because RAG1 and AID activity occurs outside of Ig genes, we assessed whether these enzymes provide the secondary genetic lesions in Emu c-myc transgenic mice to promote lymphoma development. We found that the tumor incidence and tumor phenotype in Emu c-myc transgenic mice is similar in AID+/+, AID+/- and AID-/- backgrounds in both specific pathogen-free and conventional animal facilities, indicating that AID does not contribute to lymphoma development in Emu c-myc transgenic mice. To examine the role of RAG proteins in Emu c-myc mice, we examined Emu c-myc transgenic mice that harbor the Ig-HEL heavy- and light-chain transgenes, and thus have reduced RAG expression in B cells. We found that tumor incidence was not affected by these Ig transgenes. However, we found that RAG1-/- Emu c-myc mice exhibited accelerated tumor development compared to controls. This data combined with our finding that Emu c-myc mice lived longer in the conventional facility than in the specific pathogen-free facility suggest an immune-mediated activity that suppresses lymphoma development. PMID- 18408760 TI - High-resolution, dual-platform aCGH analysis reveals frequent HIPK2 amplification and increased expression in pilocytic astrocytomas. AB - Pilocytic astrocytomas (PAs, WHO grade I) are the most common brain tumors in the pediatric and adolescent population, accounting for approximately one-fifth of central nervous system tumors. Because few consistent molecular alterations have been identified in PAs compared to higher grade gliomas, we performed array comparative genomic hybridization using two independent commercial array platforms. Although whole chromosomal gains and losses were not observed, a 1-Mb amplified region of 7q34 was detected in multiple patient samples using both array platforms. Copy-number gain was confirmed in an independent tumor sample set by quantitative PCR, and this amplification was correlated to both increased mRNA and protein expression of HIPK2, a homeobox-interacting protein kinase associated with malignancy, contained within this locus. Furthermore, overexpression of wild-type HIPK2, but not a kinase-inactive mutant, in a glioma cell line conferred a growth advantage in vitro. Collectively, these results illustrate the power and necessity of implementing high-resolution, multiple platform genomic analyses to discover small and subtle, but functionally significant, genomic alterations associated with low-grade tumor formation and growth. PMID- 18408762 TI - Methyl jasmonate binds to and detaches mitochondria-bound hexokinase. AB - Cellular bio-energetic metabolism and mitochondria are recognized as potential targets for anticancer agents, due to the numerous relevant peculiarities cancer cells exhibit. Jasmonates are anticancer agents that interact directly with mitochondria. The aim of this study was to identify mitochondrial molecular targets of jasmonates. We report that jasmonates bind to hexokinase and detach it from the mitochondria and its mitochondrial anchor-the voltage-dependent anion channel (VDAC), as judged by hexokinase immunochemical and activity determinations, surface plasmon resonance analysis and planar lipid bilayer VDAC activity analysis. Furthermore, the susceptibility of cancer cells and mitochondria to jasmonates is dependent on the expression of hexokinase, evaluated using hexokinase-overexpressing transfectants and its mitochondrial association. Many types of cancer cells exhibit overexpression of the key glycolytic enzyme, hexokinase, and its excessive binding to mitochondria. These characteristics are considered to play a pivotal role in cancer cell growth rate and survival. Thus, our findings provide an explanation for the selective effects of jasmonates on cancer cells. Most importantly, this is the first demonstration of a cytotoxic mechanism based on direct interaction between an anticancer agent and hexokinase. The proposed mechanism can serve to guide development of a new selective approach for cancer therapy. PMID- 18408761 TI - BIBW2992, an irreversible EGFR/HER2 inhibitor highly effective in preclinical lung cancer models. AB - Genetic alterations in the kinase domain of the epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC) patients are associated with sensitivity to treatment with small molecule tyrosine kinase inhibitors. Although first-generation reversible, ATP-competitive inhibitors showed encouraging clinical responses in lung adenocarcinoma tumors harboring such EGFR mutations, almost all patients developed resistance to these inhibitors over time. Such resistance to first-generation EGFR inhibitors was frequently linked to an acquired T790M point mutation in the kinase domain of EGFR, or upregulation of signaling pathways downstream of HER3. Overcoming these mechanisms of resistance, as well as primary resistance to reversible EGFR inhibitors driven by a subset of EGFR mutations, will be necessary for development of an effective targeted therapy regimen. Here, we show that BIBW2992, an anilino-quinazoline designed to irreversibly bind EGFR and HER2, potently suppresses the kinase activity of wild type and activated EGFR and HER2 mutants, including erlotinib-resistant isoforms. Consistent with this activity, BIBW2992 suppresses transformation in isogenic cell-based assays, inhibits survival of cancer cell lines and induces tumor regression in xenograft and transgenic lung cancer models, with superior activity over erlotinib. These findings encourage further testing of BIBW2992 in lung cancer patients harboring EGFR or HER2 oncogenes. PMID- 18408763 TI - Evidence of functional interaction between NuMA-RARalpha and RXRalpha in an in vivo model of acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized by reciprocal balanced chromosomal translocations involving retinoic acid receptor-alpha (RARalpha). RARalpha heterodimerizes with the retinoid X receptor-alpha (RXRalpha) and transcriptionally regulates myeloid differentiation in response to ATRA (all trans retinoic acid). Several lines of evidence suggest that APL fusion proteins interact with RXRalpha. To elucidate the role of RXRalpha in APL, we conditionally knocked out RXRalpha in the hCG-NuMA-RARalpha APL mouse model. Phenotype analysis of NuMA-RARalpha+ mice demonstrated that these mice developed a myeloproliferative disease-like myeloid leukemia within 4 months of birth. While hemizygous and homozygous RXRalpha conditional knockout mice were phenotypically normal as late as 12 months of age, we observed that the leukemic phenotype in NuMA-RARalpha+ mice was dependent on the presence of functional RXRalpha. Bone marrow promyelocyte counts were significantly reduced in NuMA RARalpha+ mice with RXRalpha knocked down. Significant differences in the accumulations of Gr-1+ and Mac-1+ cells were also seen. We further observed that genes previously identified to be cooperating events in APL were also regulated in an RXRalpha-dependent manner. We therefore propose that the APL fusion protein NuMA-RARalpha cooperates with RXRalpha in the development of leukemia in hCG-NuMA RARalpha transgenic mice and suggest a novel role for RXRalpha in the pathogenesis of APL. PMID- 18408764 TI - FLASH acts as a co-activator of the transcription factor c-Myb and localizes to active RNA polymerase II foci. AB - The c-Myb oncoprotein is a DNA-binding transcription factor with a key role in early stages of hematopoiesis. To expand our knowledge of partners cooperating with c-Myb, we performed a yeast two-hybrid screening with full-length c-Myb as bait. Here, we report FLICE-associated huge protein (FLASH)/CASP8AP2 as a novel Myb-interacting protein. We show that FLASH interacts with the DNA-binding domain of c-Myb and enhances c-Myb-dependent reporter activity and expression of endogenous c-Myb target genes. Chromatin immunoprecipitation assays revealed that FLASH and c-Myb both associate with the MYC promoter region as well as with the intronic enhancer of the c-Myb target gene ADA. Furthermore, siRNA knock-down of FLASH or c-Myb both result in a reduction of MYC and ADA expression. The co activator effect is mediated through the C-terminal part of FLASH, which binds c Myb. The FLASH-induced enhancement is comparable with the increase seen with the c-Myb co-activator p300. We find FLASH localized in discrete nuclear speckles in several cell lines, co-localized with c-Myb in active RNA polymerase II foci. These results imply a novel molecular mechanism of regulation of c-Myb activity. We propose that c-Myb cooperates with FLASH in foci associated with active RNA polymerase II, leading to enhancement of Myb-dependent gene activation. PMID- 18408765 TI - CDK1 promotes cell proliferation and survival via phosphorylation and inhibition of FOXO1 transcription factor. AB - The forkhead box O (FOXO) transcription factor FOXO1 functions as a tumor suppressor by regulating expression of genes involved in apoptosis, cell cycle arrest and oxidative detoxification. Here, we demonstrate that cyclin-dependent kinase 1 (CDK1) specifically phosphorylates FOXO1 at serine 249 (S249) in vitro and in vivo. Coimmunoprecipitation assays demonstrate that both endogenous CDK1 and ectopically expressed CDK1 form a protein complex with FOXO1 in prostate cancer (PCa) cells. In vitro protein binding assays reveal that CDK1 interacts directly with FOXO1. Accordingly, overexpression of CDK1 inhibits the transcriptional activity of FOXO1 in PCa cells through S249 phosphorylation on FOXO1. Consistent with the roles of FOXO3a and FOXO4 (two other members of the FOXO family) in cell cycle regulation, forced expression of FOXO1 causes a delay in the transition from G2 to M phase. This effect is blocked completely by overexpression of CDK1 and cyclin B1. Ectopic expression of constitutively active CDK1 also inhibits FOXO1-induced apoptosis in PCa cells. Moreover, we demonstrate that the inhibitory effect of FOXO1 on Ras oncogene-induced colony formation in fibroblasts is diminished by overexpression of CDK1. Given that CDK1 and cyclin B1 are often overexpressed in human cancers including PCa, our findings suggest that aberrant activation of CDK1 may contribute to tumorigenesis by promoting cell proliferation and survival via phosphorylation and inhibition of FOXO1. PMID- 18408766 TI - Mycoplasma infection suppresses p53, activates NF-kappaB and cooperates with oncogenic Ras in rodent fibroblast transformation. AB - Prokaryotes of the genus Mycoplasma are the smallest cellular organisms that persist as obligate extracellular parasites. Although mycoplasma infection is known to be associated with chromosomal instability and can promote malignant transformation, the mechanisms underlying these phenomena remain unknown. Since persistence of many cellular parasites requires suppression of apoptosis in host cells, we tested the effect of mycoplasma infection on the activity of the p53 and nuclear factor (NF)-kappaB pathways, major mechanisms controlling programmed cell death. To monitor the activity of p53 and NF-kappaB in mycoplasma-infected cells, we used a panel of reporter cell lines expressing the bacterial beta galactosidase gene under the control of p53- or NF-kappaB-responsive promoters. Cells incubated with media conditioned with different species of mycoplasma showed constitutive activation of NF-kappaB and reduced activation of p53, common characteristics of the majority of human tumor cells, with M. arginini having the strongest effect among the species tested. Moreover, mycoplasma infection reduced the expression level and inducibility of an endogenous p53-responsive gene, p21(waf1), and inhibited apoptosis induced by genotoxic stress. Infection with M. arginini made rat and mouse embryo fibroblasts susceptible to transformation with oncogenic H-Ras, whereas mycoplasma-free cells underwent irreversible p53 dependent growth arrest. Mycoplasma infection was as effective as shRNA-mediated knockdown of p53 expression in making rodent fibroblasts permissive to Ras induced transformation. These observations indicate that mycoplasma infection plays the role of a p53-suppressing oncogene that cooperates with Ras in cell transformation and suggest that the carcinogenic and mutagenic effects of mycoplasma might be due to inhibition of p53 tumor suppressor function by this common human parasite. PMID- 18408767 TI - PCDH8, the human homolog of PAPC, is a candidate tumor suppressor of breast cancer. AB - Carcinoma is an altered state of tissue differentiation in which epithelial cells no longer respond to cues that keep them in their proper position. A break down in these cues has disastrous consequences not only in cancer but also in embryonic development when cells of various lineages must organize into discrete entities to form a body plan. Paraxial protocadherin (PAPC) is an adhesion protein with six cadherin repeats that organizes the formation and polarity of developing cellular structures in frog, fish and mouse embryos. Here we show that protocadherin-8 (PCDH8), the human ortholog of PAPC, is inactivated through either mutation or epigenetic silencing in a high fraction of breast carcinomas. Loss of PCDH8 expression is associated with loss of heterozygosity, partial promoter methylation, and increased proliferation. Complementation of mutant tumor cell line HCC2218 with wild-type PCDH8 inhibited its growth. Two tumor mutants, E146K and R343H, were defective for inhibition of cell growth and migration. Surprisingly, the E146K mutant transformed the human mammary epithelial cell line MCF10A and sustained the expression of cyclin D1 and MYC without epidermal growth factor. We propose that loss of PCDH8 promotes oncogenesis in epithelial human cancers by disrupting cell-cell communication dedicated to tissue organization and repression of mitogenic signaling. PMID- 18408774 TI - Clinicopathological study of 100 Nigerian cases of ameloblastoma. AB - SUMMARY: Clinicopathological analysis and growth rate study of 100 cases of ameloblastoma was undertaken. The tumour had site predilection for the mandible (91.0%) and occurred more in males than in females (ratio 1:0.9). 62.0% of patients in the series were less than 20 years of age. The estimated monthly growth rate (EMGR) of ameloblastoma was observed to be slower in males although the mean tumour volume recorded in females was lower than males in the series. The biologic aggression of 4 various types of ameloblastoma was predicted using EMGR and average volume of tumour at hospital presentation as predictive markers. Solid multicystic ameloblastoma was observed with the fastest EMGR of 0.81 cm(3)/month. Peripheral ameloblastoma was observed with the slowest EMGR of 0.17 cm(3)/ month. It was concluded that all the 4 types of ameloblastoma observed were clinically distinct and showed varying biologic aggression. The solid multicystic type with the highest growth rate (0.81 cm(3)/month) was considered the most aggressive while the peripheral type (growth rate = 0.17 cm(3)/month) was the least aggressive. It is therefore necessary to distinguish between the various clinical types of ameloblastoma before a clinician proceeds with definitive treatment. PMID- 18408768 TI - Multiple pathways are involved in the anoxia response of SKIP3 including HuR regulated RNA stability, NF-kappaB and ATF4. AB - Under anoxia a coordinated, cytoprotective program is induced, called the unfolded protein response (UPR). Activating transcription factor 4 (ATF4) is a mediator of the UPR and activates a gene expression program, promoting tumour growth and survival under anoxia. A key gene induced by ATF4 under normoxic conditions is SKIP3. We characterized the induction of SKIP3 during anoxic exposure to determine whether UPR alone was sufficient or there was a more complex regulatory response to anoxia. There was temporal separation of acute hypoxia-inducible factor (HIF)-1alpha- and chronic ATF4-dependent gene expression programs. SKIP3 was regulated by chronic (48 h) rather than acute anoxia (<24 h) by a complex set of pathways and mechanisms, besides ATF4 induced by the classical UPR, there was transcriptional regulation by nuclear factor-kappa B (NF kappaB) and RNA stabilization by HuR. Temporal activation of the NF-kappaB pathway under anoxia protected cells from negative consequences of the oxygen stress and involved the canonical signalling pathways that promote IkappaBA phosphorylation and degradation, and reduced mRNA level of the inhibitory protein IkappaBA followed by the translational repression of IkappaBA. We also show that SKIP3 acts as an inhibitor of NF-kappaB and ATF4-dependent transcription under anoxia and provides a regulatory feedback loop. Repression of the survival pathway NF-kappaB by SKIP3 sensitized cells to metabolic consequences of the anoxic stress. Thus, the response to anoxia is mediated by three pathways independently of HIF, suggesting that combined therapeutic approaches would be needed to maximize effects against this pathway. PMID- 18408773 TI - The role of maintenance chemotherapy after autotransplantation for acute lymphoblastic leukemia in first remission: single-center experience of 100 patients. AB - A total of 100 adults with ALL in first CR received melphalan (110 mg/m(2)) with TBI followed by autologous marrow (n=35) or single-agent melphalan (200 mg/m(2)) followed by autologous blood stem cells (n=65). After adequate hematologic recovery, maintenance chemotherapy with 6-mercaptopurine, methotrexate and vincristine-prednisone was administered for 2 years. Six patients, all TBI recipients (P=0.001), died of toxicity. In total 70 patients received 6 mercaptopurine, 53 received methotrexate and 40 received vincristine-prednisone. The cumulative incidence of relapse at 7 years was 45%. The 7-year probabilities of disease-free survival (DFS) and overall survival were 45 and 48%. Age 30 years, >4 weeks to attain remission, and karyotypes t(4;11) and t(9;22) were associated with adverse outcome. Patients with 0 (standard risk), 1 (intermediate risk), and 2-3 (high risk) adverse features had 7-year cumulative incidences of relapse of 19, 53 and 82% (P<0.0001), and 7-year DFS probabilities of 73, 36 and 7% (P<0.0001). The 7-year probabilities of DFS for patients receiving 0, 1, 2 and 3 maintenance chemotherapy agents were 15, 29, 58 and 61% (P<0.0001). Maintenance chemotherapy intensity was an independent determinant of outcome in Cox analysis. Maintenance chemotherapy after autotransplantation reduces relapse and improves outcome in adult patients with ALL. PMID- 18408775 TI - Pattern of extremity injuries in polytrauma in Lagos, Nigeria. AB - BACKGROUND: Extremity injuries are common in polytrauma, which remains a leading cause of death, long hospitalisation and disability for all ages. There are few research works on the pattern of these extremity injuries in Lagos, Nigeria. OBJECTIVE: To describe the pattern of extremity (limb) injuries in polytraumatised patients. PATIENTS AND METHODS: This was a prospective study carried out at the Lagos University Teaching Hospital, between 1st November 2004 and 31st October 2006. RESULTS: A total of 291 patients presented with multiple injuries between November 2004 and October 2006. All of them had extremity injuries. The studied patients were aged 2 to 76 years with a mean of 31.2 +/- 16.4. Two hundred and twenty two (76.3%) of the patients were aged 40 years or less. There were 219 males and 72 females showing a male to female ratio of 3:1 Road traffic accident was the most common aetiology (73.2%). The most frequently seen fractures were tibia 27.8%, femur and fibula 24.7% each. An overwhelming majority (88.8%) had multiple fractures. The commonest fracture combinations were tibia/ fibula 24.7% and radius/ulna 19.6%. The most frequently associated injury was head injury 26.8%. Others were spinal and abdominal injuries 13.4% each. Hip dislocation accounted for a majority (56%) of all dislocations seen in this study. CONCLUSION: Lower extremity injuries were commoner than upper extremity injuries in polytrauma. Majority of them had multiple fractures especially among bones that have close anatomical location. Head injury was the most commonly associated injury. PMID- 18408776 TI - The effect of accessibility to drugs on outcome of therapy in patients with malignant lymphoma. AB - OBJECTIVE: To determine the compliance rate, reasons for default and factors affecting compliance. PATIENTS AND METHODS: A prospective study of patients with malignant lymphoma (ML) at the University of Benin Teaching Hospital, Benin City, Nigeria. A total of 190 patients on chemotherapy for ML were followed up for between 6 and 18 months during study period (1995-2003). The reasons for default were recorded. Compliant and noncompliant patients were compared in terms of survival and sociodemographic data. STATISTICAL ANALYSIS: Instat Package system for frequency counts, chi-squares and cross tabulations using Yates correction when necessary and the Fisher's exact test. RESULTS: Noncompliance rate was 63.2%. Major reasons for defaulting were high cost of drugs in 40 cases (33.3%), scarcity of drugs in 29 cases (24.2%) and side effects in 24 cases (20.6%). Compliance was significantly associated with higher levels of education, socioeconomic status, geographical abode (P<0.001) and gender (P= 0.031). Survival was found to be significantly associated with compliance in non Hodgkin's lymphoma (NHL) (P>0.001) while the relative risk (RR) was below unity in ML. CONCLUSION: The level of compliance with medical therapy is still very poor. Health education and the provision of affordable, accessible and appropriate medical therapy are required. A multidisciplinary approach to improve compliance of patients with medical therapy is advocated. PMID- 18408777 TI - Incidence and associated risk factors of postpartum depression in a tertiary hospital in Nigeria. AB - OBJECTIVE: To determine the incidence and associated risk factors of postpartum depression in a tertiary hospital in Nigeria. PATIENTS AND METHODS: A survey of 206 consecutive women who were 6 weeks postpartum was done, using the Edinburgh postnatal depression scale (EPDS), in a tertiary hospital in Nigeria. Fisher's exact test was used to determine significance between proportions . The Odd ratio and 95% Confidence Intervals were also determined. RESULTS: The incidence of postpartum depression was 27.2%. Preterm delivery was found to be a risk factor for postpartum depression in the study population (P<0.0001, OR=8.0, CI=2.9 22.1). CONCLUSION: It is recommended that the EPDS should be used routinely to screen for postnatal depression in puerperal Nigerian women and that mulltidisciplinary approach involving Obstetricians and Mental health professionals should be adopted in the screening for, and care of women with postpartum depression. PMID- 18408778 TI - Antenatal cardiotocography: experience in a Nigerian tertiary hospital. AB - OBJECTIVES: To compare selected perinatal characteristics between infants of mothers with reactive and non-reactive antenatal cardiotocography (non-stress test) results. PATIENTS AND METHODS: The medical records of all women who delivered within one week of a non-stress test were retrieved. The indication, test result, gestational age at delivery, Apgar score at 5 minutes, perinatal complications and neonatal ward admission were documented. RESULTS: One hundred and twelve women met study criteria among whom 51 tests were reactive and 61 were non-reactive. Post-datism, intra-uterine growth restriction, hypertensive disorders of pregnancy, and premature rupture of membranes were the commonest indications for the test. Post-datism was more commonly associated with a non reactive test result. Women with non-reactive tests were almost twice as likely to be delivered by emergency Caesarean section, compared with women with reactive tests; although the test result did not significantly influence the mode of delivery. A non-reactive test was significantly associated with a higher perinatal mortality (p = 0.04). Although the reactive test was associated with a three-fold reduction in the incidence of low Apgar scores compared with the non reactive test, this difference was not statistically significant (p = 0.18). A non-reactive test was significantly associated with small for gestational age infants (p = 0.01). CONCLUSIONS: Non-reactive non-stress test may be associated with higher perinatal mortality. When appropriately utilised, the test is a valuable tool for early detection of foetal compromise. Antenatal cardiotocography has a place in obstetric practice in low-resource settings for improving perinatal care. PMID- 18408779 TI - HIV/AIDS in ophthalmic patients: The Guinness Eye Centre Onitsha experience. AB - OBJECTIVES: To determine the incidence and pattern of ocular problems of HIV/AIDS at the Guinness Eye Centre Onitsha, Nigeria. METHODS: The case files of all patients who had HIV test at the Guinness Eye Centre Onitsha over a 6-year period were examined. Those who tested positive for HIV were further analysed. Information obtained included patients' demographic characteristics, clinical diagnosis, ocular and systemic co-morbidity, visual acuity and follow-up. RESULTS: Of 1011 patients, 100 (9.9%), 51 males and 49 females, were confirmed HIV-positive. The age range was 21 - 80 years; median -31 years. Fifty-five patients (55%) were or had been married; 45 (45%) were single. Herpetic eye disease constituted 50% of the cases with herpes zoster ophthalmicus accounting for 48%. Bilateral ocular disease occurred in 19 patients (19%) viz: cytomegalovirus (CMV) retinitis (6%); corneal ulcers (6%); uveitis (4%); ocular motor palsy (2%) and ocular gunshot injury (1%). Non-HIV ocular lesions occurred in 20 patients (20%) as follows: bacterial corneal ulcer (8%); globe laceration (6%); non-CMV associated rhegmatogenous retinal detachment, cataract, and secondary orbital tumour (2% each). Systemic co-morbidities were present in 10 patients (10%), namely, emaciation (6%), pulmonary tuberculosis and abdominal malignancy with orbital metastases (2% each). Twenty three patients (23%) had bilateral blindness; 45 (45%) had uniocular blindness; 73.4% of the affected eyes were blind at presentation with 25% having no light perception (NPL). CONCLUSIONS: The incidence of HIV seropositivity doubled in the hospital over nearly 10-year period. Herpes zoster ophthalmicus remains the commonest ocular manifestation although CMV retinitis is becoming common. Since 20% of the patients had non-HIV ocular lesions, eye-health workers are advised to always take universal precautions in order to prevent the spread of the infection within and outside the hospital. PMID- 18408780 TI - A study of the work and home domain characteristics of anaesthetists in Nigeria. AB - OBJECTIVES: To examine the work and home domain characteristics of anaesthetists in Nigeria, and the impact of these on work and family life. MATERIALS AND METHODS: A total of 200 self-administered questionnaires were distributed to anaesthetists in various health institutions in Nigeria. Of these, 114 (57%) were returned. Information contained in the questionnaire included demographic characteristics, qualification, rank and years spent in the specialty, spouse occupation and details of work and family life. RESULTS: The 114 respondents consisted of 86 males (77%) and 28 females (22.8%), aged between 30 and 59 years. The average weekly working hours were 58.4 hours. About half of the respondents could study anaesthesia at home sometimes, while 25% each could do so always or not at all. About 41% of the respondents could respond to emergencies at home immediately, while 45% could only do so after first reporting to work. Respondents' recommendation to enhance work and family life included reduced work load, more flexible work schedule, accommodation close to place of work, among others. CONCLUSION: Positive impact on service output as well as enhanced job satisfaction will ensue if the recommendations in this study are implemented by the relevant authorities. PMID- 18408781 TI - The in-vitro antimicrobial activity of Abrus precatorius (L) fabaceae extract on some clinical pathogens. AB - OBJECTIVES: To investigate the in-vitro antimicrobial activities of the extracts of Abrus precatorius on some clinical isolates as resistance to available and affordable antibiotics by these pathogens is on the increase. METHOD: In this study the antimicrobial effects of the extracts of Abrus precatorius from leaves, stem and the seed oil were tested against Staphylococcus aureus ATCC 25923, three clinical S. aureus isolates from different sources, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus anginosus (S.milleri), Bacillus subtilis, Corynebacterium spp (toxigenic strain of the mitis biotype), Escherichia coli ATCC 25922, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa as well as Candida albicans using the agar well diffusion technique. Aqueous and methanolic extraction, using the soxhlet extractor was carried out on all plant parts used while petroleum ether was the solvent used to extract the seed oil. To measure the MIC values, various concentrations of the stock, 512, 256, 128, 64, 32, 16, 8 and 4 microg/ml were assayed against the test bacteria. RESULT: At the different concentrations of the extracts used (512 microg/ml - 4 microg/ml), Staphylococcus aureus was the most sensitive organism with an MIC of 8 ug/ml for the leaf extract. Extract from the stem and seed oil were potent against some of the gram-positive bacteria and Candida albicans but not against S anginosus, E. faecalis and gram-negative bacteria tested. The pH of the extracts ranged between pH5 and pH8. This study demonstrates that Abrus precatorius particularly the seed oil has a potent antimicrobial activity. CONCLUSION: The results substantiate the ethno botanical use of different parts of Abrus precatorius for the treatment of various bacteria-related diseases. Topical application of Abrus precatorius extracts in ointments may be recommended especially for treating superficial infections caused by Staphylococcus aureus. PMID- 18408782 TI - Natal and neonatal teeth: a review. AB - The presence of teeth in newborn is uncommon. Sometimes it occurs, appearing in about one in every 2,000 to 3,000 live births. Natal teeth are teeth already present at time of birth, neonatal teeth erupt during the first 30 days after birth. The normal eruption of the first teeth is quite exciting but their occurrence at birth or few days after birth is quite disturbing because of societal unpleasant reaction towards it in some communities and calls for concern. The majority of natal and neonatal teeth represent the early eruption of normal primary teeth. Less than 10% of natal teeth are supernumerary. Although the majority of natal teeth are isolated incidents, their presence may be associated with some syndromes. Complications include discomfort during suckling causing irritation and trauma to infants tongue, sublingual ulceration, laceration of the mother's breasts and aspiration of the teeth. The objective of the present study was to present a review of the literature on the current information on this topic and give treatment options if necessary. PMID- 18408783 TI - Indications for extraction of third molars: a review of 1763 cases. AB - OBJECTIVE: Third molars have been described as different from other teeth in the oral cavity. They have the highest rate of developmental abnormalities and, most importantly, are last in the eruption sequence. We investigated reasons for third molar teeth extraction in patients attending the outpatient oral surgery clinic of the Lagos University Teaching Hospital, Nigeria. MATERIALS AND METHODS: A retrospective review of patients who had third molar extractions between January 2001 and June 2006 was conducted. Data retrieved and analysed were: age and sex of patients, indication for extractions, type of teeth extracted, technique of extractions (surgical and non-surgical) and angulations of impaction in case of impacted lower third molars. RESULTS: A total of 1763 patients (mean age +/-SD, 33.74+/- 13.3 years; range 15 - 92 years) had their third molars extracted during the period. Majority (58%) of them were females. Surgical extraction was carried out in 506 (28.7%) patients while 1257 (71.3%) patients had non-surgical extractions. Caries and its sequela (63.2%) was the major reason for teeth extraction followed by recurrent pericoronitis (26.3%) and periodontitis (9.2%). Only 11 (0.6%) cases were removed for prophylactic reason. Patients who had their teeth removed for caries were significantly younger than those for periodontitis (P=0.000) but older than those for pericoronitis (P=0.000). CONCLUSIONS: Caries and periodontal diseases occurring in relatively older age group were the major reasons for non-surgical extraction of third molars while recurrent pericoronitis occurring in relatively younger age was the major reason for surgical extraction of impacted third molars. Prophylactic surgical extraction of third molars is not a common practice in our environment. PMID- 18408784 TI - A histopathological analysis of thyroid diseases in Ile-ife, Nigeria. a review of 274 cases. AB - OBJECTIVE: To describe the spectrum of histopathological features of thyroid diseases; analyse the occurrence of each of the types and to compare the findings with those from previous studies done in Nigeria and elsewhere. MATERIALS AND METHODS: A retrospective study of thyroid lesions seen at the OAUTHC Histopathology Department during a 10-year period between 1988 and 1997. RESULTS: The 274 surgical thyroid specimens received during the study period came from 235 (85.8%) females and 39 (14.2%) males giving a female: male ratio of 6:1. Colloid goiter accounted for about 75% of cases with an average age of occurrence of 40.7 years. Only 1.8% showed features of toxic goiter and thyroiditis each. The adenomas constituted about 6% while carcinomas constituted about 11% of cases respectively. Adenomas occurred almost a decade earlier than carcinomas. Follicular carcinoma was the commonest thyroid cancer seen in this study. CONCLUSION: This study shows that the commonest thyroid disease in Ile-Ife is colloid goiter, which is a preventable disease. Thus, public health measures such as iodination of salt and health education are called for to reduce the occurrence of this disease. PMID- 18408785 TI - Neglected massive intracerebral abscess: an unusual cause of bilateral visual loss. AB - BACKGROUND: Bilateral visual loss following chronically raised intracranial pressure is not uncommon especially in developing countries. However, this calls for concern when the cause of the raised intracranial pressure is neglected pyogenic cerebral abscess. CASE SUMMARY: A sixteen year old male student presented to our unit with eight months history of recurrent headache associated with early morning vomiting. He developed bilateral visual loss a month prior to presentation. He had sought treatment at several herbal homes and orthodox hospitals before presentation. His cranial computed tomographic scan (CT) showed a massive peripheral contrast enhancing lesion in the frontal lobes with gross midline shift and other evidence of mass effect. He subsequently had craniotomy and excision of the mass. Intraoperative and laboratory findings confirmed the lesions to be abscess. He had ciprofloxacin and metronidazole for four weeks. He had uneventful postoperative period though his visual loss has persisted. He is being followed up in the clinic. CONCLUSION: This patient illustrates that untreated frontal lobe abscess could cause bilateral visual loss from chronically elevated intracranial pressure (ICP). Even though this should be rare in contemporary neurosurgical practice, ignorance and poverty which are rife in developing countries will be important contributory factors. Physicians working in developing countries should be aware of the symptomatology of cerebral abscess and raised ICP. They should encourage patients to do cranial CT when the features are suggestive of space occupying lesions. PMID- 18408786 TI - Typhoid intestinal disease: 32 perforations in 1 patient. AB - Typhoid intestinal perforation is a common complication of typhoid fever in our environment. The occurrence of multiple intestinal perforations that involves both small and large bowel is increasingly being seen in our practice. We report a case of 32 intestinal perforations in a child. This is the highest number of perforations seen in any 1 patient in our search of the English literature. This article details our approach to management of this type of patient. PMID- 18408787 TI - Primary hyperparathyroidism presenting with severe hypertension in a middle aged Nigerian--a case report. AB - OBJECTIVE: To draw attention to primary hyperparathyroidism as a cause of severe hypertension. RESULTS: A 47 year old Nigerian male presented with headache, occasional blurring of vision and dyspnoea on mild exertion of 2/12 duration. He had been troubled by painful osteoarthritis of the knees for 2 years for which he was taking NSAIDs. He was found to be severely hypertensive, BP 210/130mmHg and had bilateral knee crepitus. BP was resistant to control on Nifedipine R and Moduretic. Serum urea, creatinine, uric acid were normal but there was hyperacalcaemia and hyperchloraemia. Haematological indices, urinalysis, microscopy and culture of urine were normal. Parathyroid hormone level was raised. A parathyroid MIBI scan study showed an extensive area of significance towards the inferior pole of the right lobe of thyroid medially with a second area of very low significance medial to the left pole. These findings indicated the presence of a right inferior parathyroid adenoma. He had parathyroid surgery and a large adenoma in the right inferior gland and a smaller left inferior gland were removed and confirmed histologically. Corrected calcium and parathyroid hormone levels dropped to normal, and the BP was easily controlled with Lisinopril 5mg daily subsequently. He is not currently on antihypertensive medication two years post surgery. CONCLUSION: This case highlights the need for thorough investigation of cases of hypertension to exclude specifically secondary causes, which in some cases may be endocrine in origin and may easily be corrected. PMID- 18408788 TI - Isolated dextrocardia with bilateral undescended testis and bilateral polydactyl of the hands and feet: a case report. AB - The incidence of dextrocardia with or without situs inversus is rare though the true incidence in Nigeria is not known. Plain films are the preliminary investigations of choice in diagnosis of situs anomalies. The aim of this case report is to emphasise the importance of recognition of the spectrum of situs anomalies, because the altered anatomy associated with these anomalies may result in confusing imaging findings, misdiagnosis and difficulties with treatment when seen in conjunction with acquired abnormalities of the chest and abdomen. This study also aim to emphasise the need for accurate placement of radiographic markers by the radiographers before exposure of the films as the key for identification of situs anomalies. The case of dextrocardia without situs inversus diagnosed in a 4-month old baby boy who also had bilateral undescended testis and bilateral polydactyl of hands and feet is presented. The imaging modalities for assessment of situs anomalies are discussed. PMID- 18408789 TI - Early sequential findings in the genitalia of rams experimentally infected with Actinobacillus seminis. AB - AIM: To investigate and assess the sequential pathological changes in the epididymis and testis of young rams injected intra-epididymally with Actinobacillus seminis. METHODS: Twenty yearling Suffolk and Suffolk-cross rams were randomly divided into two groups comprising 16 test and four control animals. Each test ram received 2.3 x 109 cfu/ml of A. seminis injected intra epididymally. Every 24 h post-inoculation (p.i.), two test rams were randomly selected, euthanised, and necropsied, until the end of the experiment at 192 h p.i. One control animal was euthanised at 24 h, 48 h, 96 h and 144 h p.i., respectively. The reproductive tract of each ram was carefully examined, lesions photographed, and tissues cultured. Thin sections of tissue samples were fixed and examined by light microscopy; additionally, epididymal tissues were examined by scanning electron microscopy (ScEM). RESULTS: Gross lesions were observed in the cauda epididymis of all test rams, and ranged from swelling at 24 h p.i. through enlargement and granuloma formation from 72 h p.i., to gradual enlargement and increasing firmness by 192 h p.i. Gross testicular atrophy was observed in three rams. Histologically, spermatic granulomas were evident in the epididymis and the tunica vaginalis of 10 and four rams, respectively. Cauda epididymitis was present in all rams, and caput and corpus epididymitis in eight and four rams, respectively. Interstitial orchitis was observed in seven, testicular degeneration in 14, and localised and diffuse tunica vaginalitis in 12 rams. Epididymal vasculitis and infiltration of eosinophils were observed as early as 24 h p.i. Moderate disruption of the epididymal duct from 72 h p.i., with subsequent release of spermatozoa into the interstitium, was revealed by ScEM. Actinobacillus seminis was cultured from the granuloma of six test rams from 72 h p.i. CONCLUSIONS: Actinobacillus seminis has the ability to persist in the genitalia of young rams following experimental infection. Suppurative epididymitis is observed as early as 24 h p.i., and spermatic granuloma within 72 h p.i. Infiltration of eosinophils appears to be an early host response to the bacterium, and may play a role in the pathogenesis of the epididymitis. PMID- 18408790 TI - An examination of the relative reliability of laboratory case submissions in determining the prevalence of anthelmintic resistance in sheep nematodes in New Zealand, and the possible influence of test analysis methodology on such data. AB - AIM: To evaluate the likely reliability of laboratory case submissions in assessing the prevalence of anthelmintic resistance in sheep nematodes in New Zealand, and to examine the possible influence of two alternative faecal nematode egg count reduction (FECR) analysis methodologies on such data. METHODS: A comparison was made between the prevalence of anthelmintic resistance determined using faecal egg count reduction tests (FECRTs) conducted on randomly selected sheep farms in a national survey with those derived from similar case material submitted to a veterinary pathology laboratory on a more ad-hoc basis. A comparison was also made between two alternative FECR analysis methodologies using the latter data. One methodology involved a partially differentiated procedure in which FECRs for individual nematode genera were only undertaken in those instances where reductions in total strongylid faecal nematode egg counts (FECs) (excluding Nematodirus) of <95% were recorded. The other was a fully differentiated method where reductions in FECs for individual parasites were undertaken in all cases. RESULTS: Although there were some differences between them the results showed that there were considerable similarities between the prevalence data obtained from both the national survey and laboratory case submissions. This was particularly evident in relation to the overall pattern of involvement of the various nematode genera and the types of anthelmintic concerned. A comparison between laboratory case submission data analysed using a partially differentiated FECR methodology with that of a fully differentiated procedure, however, suggested that the use of the former practice was likely to lead to the 'true' prevalence of resistance being underestimated. CONCLUSIONS: The results of this study indicate that examination of FECRT case submissions to veterinary laboratories may offer a useful source of information regarding changes in the prevalence of anthelmintic-resistant sheep nematodes in New Zealand. They also lend support to suggestions that the recently completed national survey may have provided a conservative estimate of the prevalence of such resistance. PMID- 18408791 TI - Does control of bovine viral diarrhoea infection make economic sense? AB - AIM: To provide an economic analysis of the costs of control or eradication of bovine viral diarrhoea (BVD) against the estimated costs of the disease. METHODS: A decision-tree approach was adapted to an analysis of the costs of bovine viral diarrhoea virus (BVDV) infection and that of three main control options (vaccination, test-and-cull, and increased biosecurity) and their combinations, to the dairy industry in New Zealand. The model was based on an average herd of 322 milking cows. Endemic, epidemic and sporadic effects of BVDV infection were modelled in the herd, to derive an estimate of costs. RESULTS: The cost of BVDV infection to an infected average-sized dairy herd in New Zealand was estimated to be NZ $11,334 (or NZ $35.19 per cow) per annum, and NZ $48,311 over 10 years. Based on these calculations, the estimate of the annual cost of BVDV infection to the dairy industry in New Zealand was in excess of NZ $23 million per annum. While all of the control options required financial input, the rate of return compared with the cost of BVD, when viewed over a 10-year term, was as high as 123%. CONCLUSIONS: All control options offered considerable savings compared with the cost of BVD infection, and control is economically favourable. Uncertainty over the likely efficacy of the control options under field conditions in New Zealand would not allow a firm choice of one option over another at this stage, and more work on determining the efficacy of those control options in New Zealand is needed. PMID- 18408792 TI - Multiple species of nematodes resistant to ivermectin and a benzimidazole levamisole combination on a sheep farm in New Zealand. AB - AIM: To confirm the presence of multiple anthelmintic resistance on a sheep farm in New Zealand. METHODS: Three groups of 10 weaned Romney-cross lambs were treated either with an oral dose of ivermectin (0.2 mg/kg), or a benzimidazole/levamisole (BZ/LEV) combination (4.75 albendazole and 7.5 mg/kg levamisole), or were left untreated. Ten days later, animals were necropsied, and adult worms recovered and identified from the abomasa and small intestines. Pre- and post-treatment faecal nematode egg counts (FEC) were recorded, and larval cultures were performed. RESULTS: In a faecal egg count reduction test (FECRT), adjusted to reflect pre- and post-treatment larval culture results, ivermectin resistance was detected in Teladorsagia (Ostertagia), Trichostrongylus and Haemonchus spp, while BZ/LEV combination- resistant Teladorsagia and Trichostrongylus spp were also present. Adult worm counts confirmed these results, and identified the species involved as Teladorsagia circumcincta, Trichostrongylus colubriformis and H. contortus. CONCLUSION: Multiple, multi generic anthelmintic resistance was confirmed on a sheep property in New Zealand. This included the first confirmed case of ivermectin resistance in T. colubriformis from sheep in New Zealand. PMID- 18408793 TI - The effects of sex and age on phytohaemagglutinin skin-testing of deer. AB - AIM: To determine if there are sex- or age-related differences in the increase in skinfold thickness in response to the mitogen phytohaemagglutinin (PHA) in red deer. METHODS: One dose of 250 mug PHA was injected intradermally in the right side of the neck, and phosphate buffered saline (PBS) was injected at a second site as a control, in 110 (51 males and 59 females) captive Iberian red deer (Cervus elaphus hispanicus), ranging in age from 21 months to > or =5 years. Skinfold thicknesses were measured immediately before and 72 h following injection. RESULTS: There was a significant effect of gender on the average increase in skinfold thickness; males had greater increases (8.8 (SEM 0.57) mm) than females (4.23 (SEM 0.39) mm) after correcting for other confounding variables. No age-related differences were evident, but differences between sexes were more marked with increasing age. CONCLUSIONS AND CLINICAL RELEVANCE: Effects of gender, probably due to differences in energetic and reproductive constraints in red deer, should be taken into account when interpreting skinfold-test data, both in ecology and in the control of tuberculosis (Tb). Males tend to have a thicker skin than females, so skinfold increase relative to the thickness of the skin, rather than skinfold increase per se, should be used as a more appropriate measure of skinfold increase. This may also have clinical relevance in the interpretation of tuberculin skin testing. PMID- 18408794 TI - Coat-colour dilution and hypotrichosis in Hereford crossbred calves. AB - AIM: To investigate cases of coat-colour dilution and hypotrichosis in a group of Hereford x Friesian crossbred calves, and to define the underlying molecular genetics of the disorder. METHODS: The investigation was predicated on the hypothesis that this disorder was similar to a known dominantly inherited disorder of calves of black breeds crossed with Simmental cattle, for which there were candidate gene mutations. Sequence analyses of PCR amplicons from exon 1 and exon 11 of the premelanosome protein 17 gene (PMel17) were carried out. Restriction enzyme digestions of amplicons were followed using electrophoresis of digested fragments. RESULTS: It was shown that an affected calf and its Hereford sire were heterozygous for a three-base deletion in exon 1 of the PMel17 gene. These two animals were also heterozygous for a second mutation in exon 11 of the PMel17 gene. Four other related animals were likewise heterozygous for both mutations in the sire's herd of origin. CONCLUSIONS: Coat-colour dilution and hypotrichosis in Hereford crossbred calves in New Zealand is the same genetic disorder as that previously described in Simmental crossbred calves, and is linked to mutations in the PMel17 gene. PMID- 18408795 TI - Long-term assessment of pancarpal arthrodesis performed on working dogs in New Zealand. AB - AIM: To determine the outcomes following unilateral pancarpal arthrodesis (PCA) in working dogs in New Zealand, in terms of return to work and ability, as assessed by the owner. METHODS: Working dogs treated using unilateral PCA were identified by searching the medical records of the Massey University Veterinary Teaching Hospital (MUVTH) by diagnosis and breed code. Eight Heading dogs (working Collies) and four New Zealand Huntaways were identified with carpal pathology that had been treated by dorsal-plate application, bone-grafting, and casting. All dogs were actively in work on sheep or cattle farms at the time of injury. Case files and radiographs were retrospectively analysed, and the ability of the dog to work after surgery and owner satisfaction with the outcome were assessed using a questionnaire conducted at a mean follow-up interval of 5 years. RESULTS: Following arthrodesis, 6/12 (50%) dogs could perform duties as before surgery. A further four (33%) dogs could perform most former duties. Ten of the twelve owners were satisfied or very satisfied with resultant mobility and work performance of their dogs. Post-operative complications occurred in 50% of dogs, but in only one case affected the eventual outcome. Eleven owners felt surgical repair was worthwhile in a trained working dog. CONCLUSIONS: Unilateral PCA carries a good prognosis for working dogs in New Zealand to return to work, even on hill-country properties. CLINICAL RELEVANCE: This study may allow veterinarians to provide a more accurate prognosis for working dogs requiring PCA. Working dogs that have sustained severe carpal injury including hyper extension injury, luxation and fracture, or dogs with crippling carpal osteoarthrosis (OA) can return to work after PCA. PMID- 18408796 TI - Severe haemoptysis associated with seizures in a dog. AB - CASE HISTORY: A 7-year-old, sterilised male German Shepherd dog was presented with a history of three episodes of severe haemoptysis and associated dyspnoea within a 3-week period. A generalised tonic-clonic seizure was witnessed preceding the third episode. CLINICAL FINDINGS: Thoracic radiographs revealed a severe bilateral alveolar lung pattern of the caudodorsal lung fields; extension into the left cranial lung lobe was present but pulmonary vessels were within normal size limits. Frank blood was the only abnormality present at bronchoscopy. A coagulation profile, parasitological screening, thoracic and brain computed tomography (CT), and cerebrospinal fluid (CSF) cytological analysis did not identify any abnormalities. DIAGNOSIS: Haemoptysis due to either severe neurogenic pulmonary oedema or rupture of the pulmonary capillaries secondary to seizures was considered a possible diagnosis. The primary generalised seizures were attributed to late-onset idiopathic epilepsy diagnosed by exclusion of other causes. CLINICAL RELEVANCE: This is the first known case report describing severe haemoptysis associated with seizures in a dog. PMID- 18408798 TI - Porcine dermatitis and nephropathy syndrome in New Zealand. AB - CASE HISTORY: An outbreak of fatal or debilitating disease characterised by wasting and cutaneous lesions that usually necessitated euthanasia, occurred over a period of 4 months in pigs 14-16 weeks of age from a commercial 'farrow-to finish' unit in the North Island of New Zealand. During the outbreak, 67 deaths were attributed to the condition, accounting for 2-3% of total production. The outbreak preceded the diagnosis of post-weaning multisystemic wasting syndrome (PMWS) on this farm. CLINICAL FINDINGS: Sick pigs were thin, depressed, and pyrexic, and most had multiple extensive erythematous crusting cutaneous lesions that were most prominent in the region of the hindquarters, forequarters and head. Those with cutaneous lesions affecting the hindlimbs had subcutaneous oedema and enlarged superficial inguinal lymph nodes. Diarrhoea was commonly present. PATHOLOGICAL FINDINGS AND DIAGNOSIS: In addition to the skin lesions, the main gross pathological changes were renal petechiation and lymphadenomegaly. The cutaneous and renal lesions resulted from necrotising and neutrophilic vasculitis, and thrombosis and infarction of adjacent tissues. Vascular lesions were variably present in the spleen, liver, lymph nodes and intestines. Renal tubules were distended by sero-haemorrhagic fluid and proteinaceous casts. Lymphoid tissue changes typical of porcine circovirus type 2 (PCV2)- associated disease included depletion of mature lymphocytes, histiocytic infiltrations and circoviral inclusion bodies. Lesions associated with PCV2 in non-lymphoid tissues included granulomatous or non-suppurative interstitial pneumonia, interstitial nephritis, gastroenteritis, and hepatitis with hepatocellular apoptosis. DIAGNOSIS: Porcine circovirus-associated disease (PCVAD) represented by concurrent porcine dermatitis and nephropathy syndrome (PDNS) and emergent PMWS. CLINICAL RELEVANCE: This is the first published description of PDNS and of combined PDNS and PMWS in pigs in New Zealand. In addition, the prevalence of PDNS, which is usually a sporadic disorder, was inconsistent with a diagnosis of PDNS alone. PMID- 18408797 TI - Probable malignant catarrhal fever presented as transient generalised crusting dermatitis in a cow. AB - CASE HISTORY: A 2-year-old crossbred cow developed crusting ulcerative lesions that covered approximately 40% of the body. They were first observed 2 weeks after the cow calved, and were most severe over the caudal aspect of the proximal hindlegs and perineum. CLINICAL FINDINGS AND DIAGNOSIS: Generalised variably confluent 1-2-cm diameter foci of ulceration and crusting were visible. No ocular or oral lesions were visible, and the cow did not have diarrhoea. Skin biopsies revealed lesions consistent with those previously described for malignant catarrhal fever (MCF). Additionally, prominent multinucleate cells were visible. The DNA for ovine herpesvirus type 2 (OHV-2) was amplified from the skin biopsies, using PCR. The cow spontaneously made a complete clinical recovery. CLINICAL RELEVANCE: Malignant catarrhal fever should be considered in cases of ulcerative skin disease in cattle. The disease is difficult to diagnose, and a combination of skin histology as well as PCR is required. Although probably rare, it appears complete recovery from MCF is possible when the disease is confined to the skin. PMID- 18408802 TI - Recent research in science teaching and learning. PMID- 18408803 TI - A wondrous birth. AB - In this column, the author describes tending to a mother who displayed a quiet confidence throughout her pregnancy and birth experience. This birth story provides powerful support for women's inherent ability to give birth. Women already know what they need to give birth simply and easily. PMID- 18408804 TI - A tribute to sharron smith humenick: 1937-2006. AB - The purpose of this article is to honor Dr. Sharron S. Humenick, a long-time advocate of normal childbirth and Lamaze International, for her many accomplishments, her vigorous commitment to promoting the benefits of normal birth, and her passionate and diligent efforts in encouraging ongoing, evidence based research that underscores the importance of a satisfying, normal-birth experience for women and their families. Dr. Humenick was a trailblazer in life and also became an exceptional guide in demonstrating how to cope with a terminal illness and dying. Through her numerous colleagues and friends in the United States and other countries, Dr. Humenick's legacy will live on in worldwide efforts to improve the childbirth experience for women and their families. PMID- 18408805 TI - Antenatal education as perceived by health professionals. AB - This article aims to identify the learning needs of expectant and new parents for antenatal education as perceived by health professionals. In Australia, antenatal education programs are predominantly designed by health professionals, and recent research has challenged the efficacy of this approach. The data collected from 73 health professionals, reported here, demonstrates that their perceptions of the concerns and interests of expectant and new parents were divided into three interrelated categories: "need to know...what's happening," "they won't listen," and "balanced information." The health professional ideas for improving antenatal education were limited and identified a reluctance to change practice. PMID- 18408806 TI - Self-perceptions of parenting among adolescent mothers. AB - Using a descriptive correlational design, this study examined factors that contribute to self-perceptions of parenting among adolescent mothers 4 to 6 weeks postpartum. Healthy, first-time, 13- to 19-year-old adolescent mothers (n = 126) identified their own mothers as their primary source for social support. Additionally, the more tangible the support from the adolescent's mother, the lower the evaluation of self-perceptions of parenting. Older, more educated adolescent mothers were more positive in self-perceptions of parenting, suggesting that older adolescents have different needs in terms of parenting than younger adolescent mothers. These findings are important for designing and implementing nursing care and educating adolescents about the importance of social support, education, and parenting. PMID- 18408807 TI - Advocating for normal birth with normal clothes. AB - Childbirth educators need to be aware that the clothes they wear when teaching classes send a nonverbal message to class participants. Regardless of who wears the clothing or what is worn, clothes send a message; thus, both the advantages and disadvantages related to clothing choice should be considered. Ultimately, the message should reflect the values of supporting normal birth. For childbirth educators who are allowed to choose their own apparel to wear in their classes, street clothes may be the benchmark for which to strive. This article discusses the many nonverbal messages that clothes convey and provides support for the choice of street clothes as the dress for the professional childbirth educator; thus, "normal clothes to promote normal birth." PMID- 18408808 TI - The lamaze certified childbirth educator: standards of practice. AB - A student in a Lamaze Childbirth Educator Program expresses concern that some Lamaze Certified Childbirth Educators (LCCE educators) do not teach classes that reflect Lamaze standards. In this column, the ethical and professional standards of the LCCE educator and the challenges the childbirth educator experiences while practicing in the current maternity-care environment are presented and discussed. Lamaze International's Code of Ethics for Lamaze Certified Childbirth Educators provides guidance when dealing with these challenges. PMID- 18408809 TI - Addressing sexuality and pregnancy in childbirth education classes. AB - A positive, nonjudgmental, and informed approach to sexual health during pregnancy promotes acceptance of the normal functioning of women's bodies. It also encourages the development of close and supportive relationships that are so essential during pregnancy and birth. Common concerns do not need to become problems. Concerns include issues of libido, positioning, and preterm labor or fetal health, as well as myths and cultural attitudes. Childbirth educators can use tools such as the PLISSIT model to approach the topic of sexuality during pregnancy. In addition, opportunities are available in every childbirth class to acknowledge or ignore sexual issues. Perinatal educators who take responsibility for addressing this often-taboo topic can enhance women's feelings of safety and their confidence in normal birth. PMID- 18408810 TI - Research summaries for normal birth. AB - In this column, the authors summarize four research studies that further support the benefits of normal birth. The topics of the studies include the association of cesarean birth with an increased risk of neonatal death; the use of acupuncture and self-hypnosis as effective pain-management strategies; factors associated with amniotic-fluid embolism; and the positive influence of continuous support by lay doulas on obstetric outcomes for low-income women. PMID- 18408811 TI - Funny names. AB - In this column, a childbirth educator recalls Sharron Humenick's influence as the former editor of The Journal of Perinatal Education and her belief in the power of humor. The author also takes an amusing look at names parents choose for their children, one of Sharron Humenick's favorite humorous topics. PMID- 18408812 TI - [Reynaldo dos Santos and the elite's education]. PMID- 18408813 TI - [Our iceberg is melting-part 1]. PMID- 18408814 TI - [Medical productivity and the learning curve]. PMID- 18408815 TI - [Aortic arch aneurysms: three cases treated with different surgical approaches]. AB - Aortic arch aneurysms may be treated with different surgical approaches. Each case represents a complex opportunity for choosing the best available strategy. We present three different cases in which a particular surgical choice was found to be the most adequate to the specific problem. PMID- 18408816 TI - [Continuous regional analgesia in thoracic surgery]. AB - The aim of the present work was to describe the development and results of continuous regional analgesia protocols in post-operative analgesia after thoracic surgery. All patients submitted to thoracic surgery had continuous thoracic epidural or continuous thoracic paravertebral blocks with adequate analgesia without severe side effects. The program provided training of the entire staff, implementation of protocols for safe pain control, with standard policies and procedures on records of analgesia and on treatment of its side effects. The use of demanding continuous regional analgesia techniques is possible, but requires systematic assessment by trained personal, training of medical and nursing staff, clear distribution of responsibilities, use of printed indications, systematic records of pain and complications, education of patients and continuous quality control and revision of methods. PMID- 18408817 TI - [Solitary fibrous tumor of the pleura]. AB - Solitary fibrous tumors of the pleura are rare tumors originating from the mesenchymal cells of the submesothelial tissue of the pleura. In half of the cases, the neoplasm presents as an asymptomatic mass, often with quite large dimensions. It is a benign tumor in 80% of the cases and even when considered malignant has a good prognosis if totally removed. We present the case of a 78 year-old female, who was evaluated for lower respiratory infection, in whom the diagnosis of Giant Solitary Fibrous Tumor of the Pleura was made. She was submitted to surgical en bloc resection and is asymptomatic and with no evidence of recurrence in the 4th month follow-up evaluation. PMID- 18408818 TI - [Massive haemothorax in a patient with double antiplatetel aggregation therapy]. AB - The authors report the clinical case of a patient with double antiaggregant therapy who was admitted with a massive traumatic haemothorax and haemorrhagic shock in the Emergency Department of our hospital. She had an immediate thoracotomy with significant haemodynamic instability. The pos-operative outcome was good and the patient was discharged home. PMID- 18408819 TI - [New trends in the prevention of myointimal hyperplasia and anastomotic fibroplasia]. AB - Vascular reconstructive surgery has faced, since the beginning, with a complex biologic phenomenon, jeopardizing the results, which is the miointimal hyperplasia and anastomotic fibroplasia, developed at the vascular anastomosis or following balloon angioplasty, leading often to a vessel occlusion and failure of the revascularization procedure. Multiple and diversified research projects have been dedicated, along the years, to the study of the intimal mechanisms and attempts to control the phenomenon by pharmacotherapeutic agents, but the results are still far of being regarded as satisfactory. The author makes a review on the pathogenesis of the miointimal hyperplasia and anastomotic fibroplasia and proceeds to a critical analysis on the most recently introduced innovations, such as is the case of the drug eluting stents, the brachitherapy and the molecular and genetic therapies. PMID- 18408820 TI - [Surgical management of non-adherent thrombosis of inferior vena cava extended to the level of renal veins]. AB - The author reports a series of 5 patients with deep venous thrombosis extended to the inferior vena cava as a floating thrombus, who underwent a venous thrombectomy, with excellent early and long-term results. The fundamentals of this therapeutic option, the diagnosis and the surgical indications are presented and discussed. PMID- 18408821 TI - [Late results of the "simplified technique" in the surgical management of a thoracoabdominal aortic aneurysm, ten years later - clinical report]. AB - The authors report the clinical case of a dissecting thoracoabdominal aortic aneurysm, in a 49-years old female, with the diagnosis of Marfan's syndrome, who underwent surgical treatment utilizing the "simplified technique", introduced by ourselves in 1983, for the management of this most demanding situation. Reviewed ten years later, having reassumed her social and professional life, a control angio-CT disclosed the procedure of aortic reconstruction and the revascularization of the digestive and renal arteries working in excellent condition. The unique and singular reconstructive procedure here utilized and its excellent long term result, justify its presentation and divulgation. PMID- 18408822 TI - [Revascularization surgery of chronic intestinal ischemia following failure of endoluminal treatment]. AB - The endoluminal interventional has played nowadays, a significant role as an alternative to the revascularization surgery of digestive arteries, but its therapeutic efficacy and duration of results have not yet been fully demonstrated. The authors report the clinical case of a 64 years old male, suffering of abdominal angina and significant weight loss, due to a pre-occlusive ostial stenosis of the celiac axis. He underwent three angioplasty/stenting attempts for the last three years, with no success to dilate the lesion and persistence of the clinical symptoms. As a consequence of the failure of the endoluminal intervention, he underwent a conventional revascularization procedure, consisting in the implantation of an aorto-hepatic prosthetic bypass graft, with immediate resolution of the symptoms and a progressive and consistent weight gain. The authors conclude by discussing the advantages and inconvenience of these two modalities for the management of chronic intestinal ischemia. PMID- 18408823 TI - [Penetrating neck injury and occlusion of internal carotid artery. Case report and literature review]. AB - In the management of penetrating carotid injuries, the surgeon must act in an emergency way to prevent active bleeding and to restore cerebral blood flow. Preservation of the carotid circulation decreases morby-mortality in these patients. Controversial diagnostic and therapeutic approach remain when no signs of bleeding or neurological ischemia are evident. The authors report the case of a male patient submitted to an emergency operation to repair a internal carotid thrombosis consequence of a penetrating gunshot injury at the area II of the neck. We reviewed the literature related to this condition and the different methods of treatment that can be employed. PMID- 18408824 TI - [Renal arterio-venous malformation and congestive heart failure]. PMID- 18408825 TI - [Dental caries and treatment needs in adolescents from the state of Sao Paulo, 1998 and 2002]. AB - OBJECTIVE: To evaluate indicators of prevalence and severity of dental caries and treatment needs in adolescents. METHODS: Data were obtained from oral health epidemiological surveys carried out in the state of Sao Paulo with adolescents from 12 to 18 years old. Dental caries experience was assessed using DMFT Index and the need for treatment was assessed using World Health Organization criteria. Significant caries index was used to define dental caries experience of one third of the group that presented greater experience of the disease. For 12-year-old group, examinations occurred in public and private schools, in 1998 (N=9,327) and 2002 (N=5,782), while 18-year-old group was examined in their households (N=5,195 in 1998 and N=257 in 2002). RESULTS: At 12 years old, DMFT index was 3.72 in 1998 and 2.52 in 2002, whereas at 18 years old, it was 8.64 and 7.13, respectively. Significant caries index at 12 was 7.40 (1998) and 5.62 (2002), at 18 it was 15.05 and 12.19, respectively. There was an increase in the need for surface restorative care at 12 year old (p<0.0001) and of sealants at 18 year old (p<0.0001). CONCLUSIONS: There was a decrease in dental caries among adolescents and most dental treatment needs were little complex. PMID- 18408826 TI - [Relative risk for AIDS between homo/bisexual and heterosexual men]. AB - OBJECTIVE: To assess the relative risk for AIDS between men who have sex with other men and heterosexual men. METHODS: Estimates on the proportion of men who have sex with men in Brazil and AIDS data from Brazil's Information System for Notifiable Diseases, were utilized. Estimates were calculated for the relative risk (RR) for AIDS of men who have sex with men with respect to heterosexual masculine population in Brazil; state and city of Sao Paulo; and state and city of Rio de Janeiro, from 1996 to 2003. The trajectory of the RR in this period was also analyzed. RESULTS: The estimates for relative risk decreased, with a tendency to stabilize: from 34.3 to 19.3 in the entire country and from 32.1 and 6.3 in the locations analyzed. In the country in 2003, the relative risk of bisexual men in relation to heterosexual men was 16.0. The RR for exclusive homosexuals had a decreasing trajectory in all of the locations studied, but not for the bisexual population. CONCLUSIONS: In all locations, the relative risk for men who have sex with other men was higher in relation to heterosexual men. This result indicates a high and persistent vulnerability among this population. PMID- 18408827 TI - [Interobserver reliability of the International Classification of Primary Care]. AB - OBJECTIVE: The International Classification of Primary Care was developed as an attempt to overcome the limitations of the International Statistical Classification of Diseases and Related Health Problems, 10th revision, when used for primary health care. The aim of the study was to evaluate the interobserver reliability of the International Classification for Primary Care when coding reasons for health-related interruption of daily activities. METHODS: Data analyzed pertained to 801 subjects from Phase 2 of the Pro-Saude Study, involving the employees of a Rio de Janeiro university who reported having been prevented from carrying out any of their usual activities (work, study, or leisure) for health-related reasons in the two weeks prior to data collection. Health problems reported in response to an open question were separately coded by two classifiers. Interobserver reliability with respect to number of health problems was calculated by weighted kappa; for the remaining analyses (chapters and full codes), crude kappa coefficients were used. RESULTS: A total of 1,641 health problems were coded by the first classifier, and 1,629 by the second. Interobserver reliability with respect to the number of health problems coded was substantial (weighted kappa=0.94; 95% CI: 0.93;0.94). Chapter and full codes showed substantial (kappa=0.89; 95% CI: 0.88;0.90) and moderate (0.76; 95% CI: 0.76;0.78) reliability, respectively. CONCLUSIONS: The results suggest that the International Classification of Primary Care is adequate for the coding of health related reasons for interruption of daily activities. PMID- 18408829 TI - [Macula study in Stargardt's disease]. AB - PURPOSE: To evaluate de macular structural damage in Stargardt's disease by optical coherence tomography, correlating with visual acuity and disease duration. METHODS: Patients with Stargardt's disease were included and submitted to visual acuity (logMAR) measurement and complementary examinations performed were color fundus photographs, fluorescein angiography and optical coherence tomography. All cases were reexamined for diagnostic confirmation and the duration of symptoms was determined. The control group was composed of the same number of subjects, matched by sex and age, without any ophthalmologic alteration. RESULTS: The sample was composed of 22 patients (44 eyes) with Stargardt's disease, 11 (50%) males and 11 (50%) females. The duration of the disease varied from 3 to 21 years (mean of 11.4 +/- 5.3 years). The groups did not show significant differences in age (p= 0.98) and sex. Concerning the macular thickness in optical coherence tomography, the variation in the study group differed significantly from the control group, presenting smaller values of thickness (p<0.001). There was negative and significant correlation between the duration of disease and the macular thickness assessed by optical coherence tomography (r=-0.57 and p=0.005). There was positive correlation between the duration of the disease and the visual acuity (r=0.50 and p=0.0167) and negative correlation between the visual acuity and the macular thickness in optical coherence tomography (r=-0.83 and p=0.0001). CONCLUSION: It was evidenced that patients with Stargardt's disease have a thinner macular thickness when compared to normal subjects, and this reduction is related to the duration of symptoms of the disease. Additionally, the thickness and also the duration of the disease influence the visual prognosis of the patients. PMID- 18408830 TI - [Positive corneoscleral rim culture in penetrating keratoplasty at the Porto Alegre Clinical Hospital]. AB - PURPOSE: To determine the incidence of positive corneoscleral rim cultures preserved in Optisol GS medium, to identify pathogens involved and possible recipient eye infection. METHODS: A hundred sixty-three corneoscleral rim cultures penetrating keratoplasties performed from January 2001 to January 2003 in the Hospital de Clinicas de Porto Alegre were reviewed. Enucleations and corneal storage were done as aseptic as possible and gentamicin 0.3% was instilled. Corneoscleral rim was divided into two segments, half was inoculated into Sabouraud broth and the other half into thioglycolate broth; inoculation into blood agar, chocolate agar and MacConkey agar was done later if necessary for pathogen identification. The receiver's eye data were reviewed. RESULTS: There were eleven positive cultures (6.7%) out of 163 evaluated corneoscleral rim cultures. Of these, four were Staphylococcus epidermidis, one was Staphylococcus aureus, one was Serratia sp., one was Pseudomonas aeruginosa and the other four were different subtypes of Candida (two Candida sp., one Candida albicans and one Candida parapapilosis). All pathogens were resistant to gentamicin. None of the eleven cases of positive corneoscleral rim cultures resulted in ocular infection at the receiver's eyes (six months follow-up). CONCLUSIONS: We found low rates of positive corneoscleral rim cultures after penetrating keratoplasty at the Porto Alegre Clinical Hospital. The most frequent involved pathogens were Staphylococcus sp and Candida sp. Although we did not identify any postoperative infection at the receiver's eyes, we recommend corneoscleral rim culture for guidance of postoperative infection, a rare but possible devastating ocular event. PMID- 18408831 TI - Tear film and ocular surface alterations in chronic users of antiglaucoma medications. AB - PURPOSE: Tear film can be altered by chronic medications that may disrupt the equilibrium responsible for the functioning of the lacrimal gland and ocular surface. The purpose of this study was to determine if antiglaucomatous chronic treatment induced alterations in the tear film and ocular surface. METHODS: After informed consent, 21 patients using antiglaucomatous eye drops for more than 8 months and 20 age- and sex-matched volunteers without eye and systemic medications (control group) were enrolled. The data of ocular discomfort, fluorescein and lisamine green staining, tear film break-up time and Schirmer test were collected and compared by Student's t test. The impression cytology data were graded and compared by chi-square test. RESULTS: Patients chronically using antiglaucomatous medications presented with significant higher fluorescein staining (p=0.003), lisamine green staining (p=0.02) and lower TFBUT (p=0.001). The other compared parameters, including impression cytology were similar between the treated and control group (p>0.05). CONCLUSIONS: The present study shows that the tear film and the ocular surface are altered in patients under antiglaucomatous medications. In common, all medications were preserved with benzalkonium chloride. Efforts to minimize the adverse effects of chronic use of antiglaucomatous drugs must be addressed. PMID- 18408832 TI - [Amniotic membrane for ocular surface reconstruction after conjunctival squamous cell carcinoma resection]. AB - PURPOSE: This study was designed to evaluate the use of human amniotic membrane for ocular surface reconstruction after conjunctival squamous cell carcinoma resection. METHODS: Amniotic membrane was obtained at the time of cesarean section and was preserved at -80 masculineC in glycerol and cornea culture media at a ratio of 1:1. The inclusion criteria were patients presenting proliferating lesions suggestive of squamous cell carcinoma (flat or elevated white lesions resembling "fish meat") that involve the conjunctiva, limbus and cornea. Eight eyes of 8 patients with conjunctival "squamous cell carcinoma" underwent tumor resection with amniotic membrane transplantation. Three of these cases underwent total corneal epitheliectomy and amniotic membrane transplantation associated with limbal autograft. RESULTS: Mean follow-up time was 17.8 months (range, 10-35 months). In four patients (71.4%) surgical treatment was successful, with good ocular surface stability. In two patients (28.6%) results were partially successful, with mild cicatricial alterations. One patient was excluded from the study due to aggressive tumor recurrence with intraocular invasion that needed to be removed with exenteration. CONCLUSION: This study suggests that amniotic membrane transplantation is a good alternative for ocular surface reconstruction after conjunctival squamous cell carcinoma resection. PMID- 18408833 TI - [Marketing role of corneal graft tissue donation to an eye bank and donors' socioeconomic profile]. AB - Penetrating keratoplasty has been the leading and the most successful type of transplant in the world, however corneal deficiency is a commom problem usually presented to corneal surgeons. PURPOSE: Impact evaluation of the number of corneal graft donations to the Sorocaba Eye Bank after the implementation of a corneal graft procurement system; to draw the socioeconomic profile of corneal graft donors of the Sorocaba Eye Bank (SEB). METHODS: Retrospective study on donations to SEB from its creation and after the development of media marketing. Prospective analysis of the socioeconomic profile of corneal graft donors by a questionnaire sent as letters to the families of the donors in a certain month. RESULTS: SEB began its work in 1971 by spreading need of organ donation through lectures in churches, shopping malls, community meetings, radio programs, television programs, etc. In the 70s, the number of retrieved corneal grafts was 1 or 2/month. Between 1984 - 1989 a procurement coordination team was trained to act in mortuaries and by 2000 they also began to work in public hospitals. In 1984 only 260 corneal grafts were retrieved. This number has been increasing to 2,778 corneal graft donations in 2004. The questionnaire was answered by 76 of the 93 donor families, with a response rate of 81.7%. Donor age had a mean of 65.1 +/- 14.7 y/o, forty-two (55.3%) were men. Educational level of the donor families was an important factor for organ donation, once 36.8% had concluded high school and 34.2% completed university. The great majority, sixty-three (82.9%) of the corneal grafts were donated through the efforts of the procurement coordination team. CONCLUSION: The role of the media and institutional credibility are mandatory for public commitment to organ donation. The proficiency of the procurement coordination team requires intensive training, as the results show that 82.9% donations were made thanks to their efforts. PMID- 18408834 TI - [Margin reflex distance measure by computerized image processing in rigid contact lens wearers]. AB - PURPOSE: To measure the MRD (margin reflex distance) in rigid contact lens wearers and controls by a new method, based on computerized image processing. METHOD: The patients were selected from the Contact Lens Sector of the Ophthalmology Service at the "Complexo Hospitalar Santa Casa de Porto Alegre", and they were divided into two groups: the first was formed of rigid contact lens wearers (63 eyes) and the second of patients without previous history of contact lens wear (30 eyes). All patients were photographed with a digital camera (Nikon Coolpix 4300). The margin reflex distance was measured by a computerized image processing using the Image J program. The study excluded patients that underwent any kind of intraocular or eyelid surgery, patients with congenital ptosis and patients with giant papillae conjunctivitis. RESULTS: The method utilized to measure margin reflex distance seems simple and more accurate. The average value of the margin reflex distance in the case group was 2.46 mm and in the control group 2.72 mm. The study shows that there is a tendency of decreasing the margin reflex distance with contact lens wear although the data were not statistically significant (p=0.22). The margin reflex distance values show a greater variability in the case group (41.46%) than in the control group (28.96%), that is more homogeneous. CONCLUSION: This study introduced a new method to measure the margin reflex distance using computerized image processing. This method is accessible and could help in follow-up of the margin reflex distance in contact lens wearers, specially those rigid. PMID- 18408835 TI - [Evaluation of hypertensive retinopathy through the oscillatory potentials of the electroretinogram]. AB - PURPOSE: To evaluate the behavior of the scotopic Oscillatory Potentials (OP) of total field electroretinogram (ERG) in hypertensive retinopathy. METHODS: Forty four patients (n=44) were submitted to clinical evaluation and subdivided in to 2 groups: hypertensives (HT) and normotensives (NT). The hypertensives patients were maintained under placebo during the period of the study. Soon afterwards, they were submitted to ophthalmological evaluation and accomplishment of ERG. Total field electroretinogram (ERG), with recording of the answers: scotopic, maximum scotopic, OP scotopic, photopic and flicker.For analysis of the OP answer the latency of the first 2 peaks and the average value of the width of the first 3 peaks of the compound of 3 consecutive answers, denominated Oscillatory Index (OI) were considered. RESULTS: The sample was composed of 44 patients, with a mean age of 51.55 +/- 7.2 range (34 to 68) years, 24 being females. Arterial hypertension affected 26 (59.1%) of the patients, while 18 (40.9%) were normotensives. The average of the obtained IO was 257.41 microv in the NT group and of 217.81 microv in HT (p=0.006). The averages of latencies obtained for peaks 1 (NT-18.42 and HT-17.91) and 2 (NT-24.54 and HT - 24.29) were not different between the groups (p>0.05). CONCLUSIONS: The hypertensive patients presented significantly smaller oscillatory index than the normotensives, suggesting that arterial hypertension might cause dysfunction of the internal retina. PMID- 18408836 TI - [Clinical characteristics of 64 individuals carrying active posterior presumptively toxoplasmic uveitis, in Pernambuco]. AB - PURPOSE: To describe clinical characteristics of posterior active uveitis presumptively by Toxoplasma gondii (PAUPT) in patients with typical lesion. Tranversal study. METHODS: Sixty-four patients with retinochoroiditis scatter and active satellite lesions examined in Pernambuco, Brazil. All were older than 10 years and immunocompetent. Gender, age, skin color, and residence were recorded. Previous uveitis, visual accuracy, intraocular pressure (IOP), and ocular examination were analyzed. RESULTS: 52% were males, most of them with white skin (68.8%). Mean age 29 years (+/-10.87). Eighty-four percent of the patients lived in the metropolitan area. 56.2% were having the first episode of uveitis. In the damaged eye, visual accuracy mean was 20/200, IOP mean 14.5 mmHg (+/-64). Hyperemia of the conjunctiva was observed in 29.7% of the patients and alterations of the cornea in 51.6%. There were cells in the aqueous humor in 62.7%. 6.2% had posterior synechiae. All had vitreous damage and 45.3% retinal vasculitis. In 42.2% of the patients, lesions were located in zone I of Holland and 90.6% had the size of one discus diameter or greater. Neuritis was observed in 28.2%. Uveitis was more frequent in the right eye (54.7%). CONCLUSION: PAUPT affects young people and the main symptom was reduction of visual acuity. IOP mean was normal. Alterations of the vitreous were observed in all cases. Injuries were equal to one discus diameter or greater and located in zone I of Holland. PMID- 18408837 TI - Efficacy and safety of green laser photocoagulation for threshold retinopathy of prematurity. AB - PURPOSE: To report the efficacy and safety of green laser photocoagulation for threshold retinopathy of prematurity (ROP). METHODS: We reviewed the clinical records of the neonates who had undergone green laser photocoagulation for threshold ROP at the Federal University of Pernambuco in Brazil between January 2004 and January 2006. All procedures were conducted with local anesthetic drops. The neonates were monitored throughout the procedure by a neonatologist. A frequency-doubled solid state laser, diode-pumped, with 532 nm wavelength was used. The presence of tunica vasculosa lentis or cataract were excluded before laser treatment. The following preoperative data were obtained for each patient: age, birth weight and the grade of ROP. Postoperative data included complications associated with the laser treatment, grade of ROP and evaluation whether further surgery was necessary due to failure of laser photocoagulation. RESULTS: Twenty two neonates underwent photocoagulation with green laser for threshold ROP. A total of 31 eyes were included in the study. The mean gestational age was 30 +/- 3 weeks and the mean birth weight was 1120 +/- 490 g. Regression of the disease after laser therapy was observed in 30 eyes (96.7%). Despite treatment one eye presented stage 4A. Only 7 eyes required repetitive laser therapy. No adverse effects such as burning anterior segment tissues or bleeding in the anterior chamber occurred. No posterior segment side-effects were observed. Cataract formation was not observed at the last follow-up examination. CONCLUSIONS: Green laser photocoagulation remains an effective and safe alternative to red laser photocoagulation and to cryotherapy in the treatment of threshold ROP. PMID- 18408838 TI - [Cataract-operated individuals at the Center-Western zone of Sao Paulo state: populational survey]. AB - PURPOSE: To determine the outcomes and causes of visual impairment and blindness in cataract-operated patients who are living in the central-western zone of Sao Paulo state. METHODS: A transversal, observational, systematic study was done involving patients inhabitants of five cities from a central-western zone of Sao Paulo state. The random sample was based on the Demographic Census Data (IBGE, 1995). All patients underwent to visual screening and complete eye examination. The individuals who had cataract-operated eyes were separated for the present study. The results were statistically analyzed by descriptive methods, frequency of occurrence and concordance proportion with 95% confidence intervals. RESULTS: At examination 2.37% of the subjects had cataract-operated eyes. Of the 201 operated eyes 26.9% had visual acuity compatible with blindness or visual impairment. The visual acuity persisted <0.3 even with spetacles in 19% of the patients. Twenty-seven and 9% of the patients had visual acuity improved with refractive error correction. The main causes of poor vision were refractive error, posterior capsule opacification (19.4%), bullous keratophaty (8.3%), cicatricial chorioretinitis (8.3%), aphakic eyes (8.3%), age-related macular degeneration (5.5%), leukoma (5.5%), glaucoma (5.5%), optic atrophy (5.5%), retinal detachment (2.8%), retinal pigment epithelium atrophy (2.8%) and high myopia (2.8%). CONCLUSION: Cataract projects are effective in reducing preventable blindness caused by lens opacity. However, long-term scheduled evaluation of operated patients is necessary avoiding consequent blindness resulting from neglected follow-up. PMID- 18408839 TI - [Corneal allograft rejection: topical treatment vs. pulsed intravenous methylprednisolone - ten years' result]. AB - PURPOSE: To evaluate the efficacy of intravenous 500 mg methylprednisolone in addition to topical treatment with 1% prednisolone in the treatment of the first episode of corneal endothelial rejection in patients that were submitted to corneal allograft transplantation. METHODS: Retrospective case-control study with 81 patients that presented the first episode of corneal endothelial rejection and were treated within the first 15 days of the onset of symptoms. RESULTS: 67 patients were treated with 1% topical prednisolone acetate and pulsed intravenous methylprednisolone 500 mg at the diagnosis of corneal allograft rejection. Fourteen patients were submitted to topical treatment only, thus forming the control group. Forty-one of 67 patients (61.2%) that were submitted to pulsed steroid had good outcome and 26 (38.8%) presented corneal graft failure while only 4 of 14 patients (28.57%) that received only topical steroids evolved with clear grafts and the remaining 10 patients (71.43%) with graft failure. Chi square showed statistically significant association (p<0.05) to greater success with pulsed methylprednisolone. CONCLUSIONS: This study suggests that the use of 500 mg intravenous methylprednisolone in addition to 1% topical prednisolone acetate for the treatment of endothelial corneal allograft rejection presents better outcomes in reverting corneal allograft rejection when compared to isolated use of 1% topical prednisolone acetate. PMID- 18408840 TI - Anterior ischemic optic neuropathy associated with metabolic syndrome. AB - PURPOSE: Metabolic syndrome denotes a common cluster of naturally connected risk factors including obesity, elevated blood pressure, insulin resistance, dyslipidemia, proinflammatory state and prothrombotic state. Anterior ischemic optic neuropathy is an acute ischaemic disorder of the optic nerve head and may lead to severe visual loss. METHODS: We considered three patients with moderate degree of diabetic retinopathy and anterior ischemic optic neuropathy. They were submitted to endocrinological examination and the diagnosis of metabolic syndrome was established. RESULTS: Cardiological examination revealed that blood pressure control was not optimal. The signs of left ventricular hypertrophy and diastolic dysfunction were confirmed by echocardiography. They are possible markers of preclinical cardiovascular disease. CONCLUSION: We observed that a variety of well-known risk factors in metabolic syndrome may be involved in serious eye and cardiological complications. The early diagnosis and treatment of these patients can not only improve visual function but also prevent cardiovascular complications. PMID- 18408841 TI - Applicability of the 2001 revised diagnostic criteria in Brazilian Vogt-Koyanagi Harada disease patients. AB - PURPOSE: To determine the applicability of the international revised diagnostic criteria for Vogt-Koyanagi-Harada disease. METHODS: Retrospective study. Medical charts of 140 patients with the diagnosis of Vogt-Koyanagi-Harada disease, from the Uveitis Sector of the Federal University of Sao Paulo (UNIFESP), were revised and classified following the revised diagnostic criteria. RESULTS: Of the 140 patients, 12.85% fulfilled the criteria for complete disease, 29.28% incomplete disease, 28.57% "probable" Vogt-Koyanagi-Harada disease and 28.27% were considered not Vogt-Koyanagi-Harada disease. CONCLUSION: The authors consider that the international revised diagnostic criteria have good applicability and are very useful to help in the diagnosis of Vogt-Koyanagi-Harada disease. PMID- 18408842 TI - [Contrast sensitivity in deep anterior lamellar keratoplasty versus penetrating keratoplasty]. AB - PURPOSE: To compare the measurement of contrast sensitivity at a distance in patients submitted to penetrating keratoplasty versus patients submitted to deep anterior lamellar keratoplasty for keratoconus treatment. METHODS: Contrast sensitivity of fourteen subjects submitted to penetrating keratoplasty and fourteen subjects submitted to deep anterior lamellar keratoplasty have been analyzed through the Functional Acuity Contrast Test (F.A.C.T) 301. RESULTS: There was no statistically significant difference between the measurements of either penetrating keratoplasty or deep anterior lamellar keratoplasty. CONCLUSION: Contrast sensitivity was similar among the subjects submitted to penetrating keratoplasty and to deep anterior lamellar keratoplasty. PMID- 18408843 TI - [Analysis of the nerve fiber layer using GDx in pseudophakic patients with posterior capsular opacification]. AB - PURPOSE: To evaluate if posterior capsule opacification (PCO) would alter the analysis of nerve fiber layer using the scanning laser polarimetry (GDx). METHODS: Pseudophakic non-glaucomatosous eyes (37 patients) underwent nerve fiber layer analysis using GDx before and after posterior capsulotomy with Nd:YAG Laser. RESULTS: In five eyes, GDx did not obtain acceptable images using the device's parameters of acquisition. In the others, mean variable values did not reach significant difference before and after posterior capsulotomy. CONCLUSION: Moderate degrees of PCO and/or acceptable images in pseudophakic patients do not alter the analysis of nerve fiber layer by GDx. Only intense degrees of PCO that hinder analyzable images make the examination impracticable. PMID- 18408844 TI - [Using of Cells Analyser software in the study of image of corneal specular microscope endothelial samples]. AB - PURPOSE: To describe differences between number of cells, evaluated endothelial area and relative error comparing endothelial samples from one endothelial image with endothelial samples obtained by guidance of a specific software, with number of cells and images as many as necessary. METHODS: A transversal study was performed, comparing the endothelial samples with and without intervention of the software. The sample consisted of 157 eyes. They were divided into 2 groups. Group 1 consisted of data of the first image and group 2 consisted of data from as many images as necessary for a complete examination guided by the Cells Analyser software (95% confidence interval and 0.05 relative error). The evaluated data were number of counted cells, equivalent evaluated field (mm2) and relative error. Student's t test with 99% confidence interval (p<0.01) was used. RESULTS: The compared data showed statistically significant differences between groups 1 and 2 in cell count (p=4x10-24), evaluated endothelial field (p=2x10-18) and relative error (p=1x10-21). CONCLUSION: The study of the sample data with and [corrected] without intervention of the Cells Analyzer software showed them to be significantly different with greater samples and smaller sample errors, evidencing the importance of their use in order to obtain specular microscopic cornea tests with small errors (less than 5%), giving them representativeness and reproducibility [corrected] PMID- 18408845 TI - [The impact of photorefractive excimer laser keratectomy (PRK) and laser in situ keratomileusis (LASIK) on visual quality and life in patients with ametropias]. AB - PURPOSE: To evaluate the quality of life, vision and stress before and after refractive surgery. METHODS: This is a longitudinal, observational study, where 100 patients were evaluated; 54 espectacles wearers, 21 contact lens users before surgery and 25 controls wearers spectacles or contact lenses, who did not want to undergo refractive surgery during one year despite refractive error. The applied questionnaires were Self Reporting Questionnaires SRQ-20 of quality of life and vision and National Eye Institute Visual Function Questionnaire NEI VFQ-25 for the assessment of mental health. The intervention group answered the questionnaires before surgery, three, six and twelve months after follow-up and the control group answered the questionnaires at six and twelve months after the baseline. The questionnaires of the intervention group were applied by an independent person. RESULTS: In the intervention group (54 spectacle wearers), 39 were treated by photorefractive excimer laserkeratectomy (PRK) and 15 by laser in situ keratomileusis (LASIK), of the 21 contact lens users: 12 received photorefractive excimer laser keratectomy and nine were treated by laser in situ keratomileusis (LASIK). The control group remained stable during the study. After three months of follow-up the intervention group showed improvement in quality of life, vision and reduction of stress. After one year of follow-up the assessments of quality of life and mental health were similar to the control group. After three months there was a significant reduction of psychiatric symptomatology in the intervention group. CONCLUSION: Patients operated for correction of ametropia showed a significant improvement in quality of life and mental health assessments. PMID- 18408846 TI - [Visual performance in myopic correction with spectacles and soft contact lenses]. AB - PURPOSE: To evaluate the visual performance by high contrast visual acuity, contrast sensitivity and wavefront in myopic patients with or without astigmatism corrected with spectacles and with three different soft contact lenses [Acuvue 2 (Vistacon J&J Vision Care Inc., USA), Biomedics 55 (Ocular Science, USA), and Focus 1-2 week (Ciba Vision Corporation, USA)]. METHODS: An interventional prospective clinical trial studied a sample of 40 myopic patients (-1.00 to -4.50 sph, with or without astigmatism up to -0.75 cyl). Each patient had one eye randomized to visual performance evaluation. RESULTS: The Zywave aberrometer detected a over refraction and significant difference between Acuvue 2 and Biomedics 55 regarding spheric refractive components and spheric equivalent. Both soft contact lenses showed hypercorrection as compared to Focus 1-2 week. Visual performance was not significantly different with spectacles and the three soft contact lenses in visual acuity and contrast sensitivity measurements. The wavefront analysis detected a significant difference in a third order aberration with and without soft contact lenses, with better visual performance with Acuvue 2 and Biomedics 55. CONCLUSION: In visual performance evaluation with spectacles and soft contact lenses the wavefront analysis was a more sensible measurement of visual function when compared to high contrast visual acuity and contrast sensitivity. The evaluation model of visual performance with wavefront analysis developed in this investigation may be useful for further similar studies. PMID- 18408847 TI - Isolated low grade pilocytic astrocytoma of the optic nerve in the elderly: case report. AB - A 68-year-old man presented with a history of a right optic glioma. Eighteen months ago he underwent a lateral orbitotomy at another institution for removal of an optic nerve mass. At that time histology revealed that the tumor was an optic nerve glioma with a pilocytic pattern. No further treatment was instituted and one year after surgery he noticed that his right eye was proptotic again. Magnetic resonance imaging of the orbit showed that his right orbit was almost completely filled with a mass which extended through the optic canal to the chiasma. The tumor was excised by a combined neurosurgical and orbital approach. Histology proved that the neoplasm was a low grade pilocytic astrocytoma of the optic nerve. PMID- 18408848 TI - [Bilateral amaurosis due to cryptococcus meningoencephalitis: case report]. AB - Cryptococcal meningitis is caused by the yeast Cryptococcus neoformans. Two varieties are recognized: var. gattii and var. neoformans. It is usually associated with immunosuppressive states, particularly HIV infection. Cryptococcal infection of the central nervous system is uncommon in immunocompetent children and difficult to diagnose. Ocular complications are common. Optic disc swelling was found in 33%. The following report describes a case of meningitis caused by C. neoformans var. gattii in an 8 year-old immunocompetent child who developed optic atrophy. The patient was treated with amphotericin B associated with corticosteroids. Possible therapeutic strategies aimed at reducing visual loss in cryptococcal meningitis have great importance to avoid this important morbidity. PMID- 18408850 TI - [Canaliculitis: case report and management]. AB - Canaliculitis is a rare infection of the lacrimal canaliculus with Actinomyces israelii as its most common etiologic agent. The objective of this paper is to report a rare pathology, as well as its clinical and surgical management. Female patient, 44 years old, presenting clinical manifestations compatible with canaliculitis, was submitted to surgical treatment preserving the lacrimal punctum. Intubation was performed into the lacrimal channel with silicone tube for three months, associated with periodical pre-programmed bathings with antibiotic, in order to avoid complications and recurrence. This case confirms the literature reports in which the surgical treatment is more effective than the conservative treatment. PMID- 18408849 TI - [Serous retinal detachment in paraganglioma: case report]. AB - The authors describe a case of a pregnant patient with arterial hypertension that resists to the treatment and retinal bilateral serous detachment. It was confirmed to be a paraganglioma by anatomicopathological examination. PMID- 18408851 TI - [Nasolacrimal stent implantation: report of a case]. AB - Epiphora (watering eye) is generally caused by insufficient drainage of tears. The most common cause of epiphora in adults is idiopathic inflammatory obstruction of the nasolacrimal duct. The traditional surgical treatment of nasolacrimal duct obstruction is an external dacryocystorhinostomy, which has an 85% to 95% success rate. To eliminate cutaneus wounds and scarring some techniques were introduced, for example the endonasal laser dacryocystorhinostomy and the nasolacrimal stent. The aim of this report was to evaluate the first case of nasolacrimal stent implantation in Brazil. The procedure was performed in the "Hospital das Clinicas" of the University of Sao Paulo - USP. A female patient with tearing of the right eye and secretion was submitted to a nasolacrimal stent implantation, with fluoroscopic guidance. The stent used in this procedure was the polyurethane Tearleader stent set. (Dr. Wilhelm type-PBN MEDICALS - Denmark). After 3 months, the patient started complaining of tearing, so the stent was removed and the patient was submitted to an external dacryocystorhinostomy. At present the patient does not have any symptoms or complaints. This procedure is less invasive and simple, causes no facial scars and avoids surgical trauma, but the long term success rates achieved using polyurethane nasolacrimal stents are low as compared with the external dacryocystorhinostomy. In addition, the patients usually complaint of tearing even having patent lacrimal system. In summary, long-term studies are needed to resolve some complications. Maybe a new stent design and new methods of unblocking the stent in situ would improve in the near future the levels of patency that currently are modest. PMID- 18408852 TI - Postoperative Alcaligenes xylosoxidans endophthalmitis: report of two cases. AB - We report two cases of postoperative Alcaligenes xylosoxidans endophthalmitis. A 78-year-old woman in good general health developed A. xylosoxidans endophthalmitis one month after an uncomplicated phacoemulsification procedure with posterior chamber intraocular lens. It was performed on her left eye, at another institution. Removal of the intraocular lens and capsule was performed because of recurrent inflammation after vitrectomy and intravitreal antibiotic injections. Her son, a 55-year-old man, developed A. xylosoxidans endophthalmitis in his left eye, two months after an uncomplicated phacoemulsification procedure with posterior chamber intraocular lens (it also was performed by the same surgeon). He also required vitrectomy with removal of the intraocular lens and capsule because of persistent disease. A. xyloso - xidans can cause chronic low grade and progressive endophthalmitis after cataract extraction that is often resistant to corrective antibiotic therapy. This kind of case is rare. There have been only previously reported seven cases worldwide to the best of our knowledge. PMID- 18408853 TI - [Leigh syndrome: case report]. AB - The authors describe for the first time in the Country a case of a 10-year-old female child, assisted at the Ophthalmology Clinic of the Hospital Universitario Clementino Fraga Filho UFRJ, with Leigh's syndrome that is part of a metabolic disease group known as mitochondrial encephalomyopathies. It is an hereditary disease transmitted by a different mode of inheritance: mitochondrial, X-linked recessive and autosomal recessive. The beginning of clinical manifestations is varied and occurs usually in the first two years of life, with progressive and insidious evolution and exacerbation periods. Diagnosis is difficult because pleomorphic presentation, based on clinical findings and complementary study related to mitochondrial production of ATP and cytochrome c oxidase deficiencies. Considering that there is no specific treatment, this is based on a palliative procedure. So, the identification of this syndrome is very important to keep it under control, since its evolution is progressive. PMID- 18408854 TI - [Vision measurement and psychophysical tests]. AB - Vision measurement is the basis for the study and standardization of visual sciences. Measurement of visual acuity has great value for research and for clinical practice. This paper (1) reviews the fundamental concepts to understand visual sense and the measuring units; (2) presents the fundamental limits to visual performance and the principles of aberration measurement of the eye; and (3) discusses methods for measuring and classifying vision with new technologies. PMID- 18408855 TI - [Letters to the editor]. PMID- 18408856 TI - [Editorial]. PMID- 18408857 TI - [Speech fluency developmental profile in Brazilian Portuguese speakers]. AB - BACKGROUND: speech fluency varies from one individual to the next, fluent or stutterer, depending on several factors. Studies that investigate the influence of age on fluency patterns have been identified; however these differences were investigated in isolated age groups. Studies about life span fluency variations were not found. AIM: to verify the speech fluency developmental profile. METHOD: speech samples of 594 fluent participants of both genders, with ages between 2:0 and 99:11 years, speakers of the Brazilian Portuguese language, were analyzed. Participants were grouped as follows: pre-scholars, scholars, early adolescence, late adolescence, adults and elderlies. Speech samples were analyzed according to the Speech Fluency Profile variables and were compared regarding: typology of speech disruptions (typical and less typical), speech rate (words and syllables per minute) and frequency of speech disruptions (percentage of speech discontinuity). RESULTS: although isolated variations were identified, overall there was no significant difference between the age groups for the speech disruption indexes (typical and less typical speech disruptions and percentage of speech discontinuity). Significant differences were observed between the groups when considering speech rate. CONCLUSION: the development of the neurolinguistic system for speech fluency, in terms of speech disruptions, seems to stabilize itself during the first years of life, presenting no alterations during the life span. Indexes of speech rate present variations in the age groups, indicating patterns of acquisition, development, stabilization and degeneration. PMID- 18408858 TI - [Performance of Brazilian Portuguese and English speaking subjects on the Test of Early Language Development]. AB - BACKGROUND: application of an American early language test in Brazilian Portuguese-speaking subjects. AIM: to compare the performance of normal Brazilian Portuguese and American English-speaking subjects on the Test of Early Language Development - Third Edition (TELD-3). METHOD: participants of this study were 120 subjects of both genders, ranging in age from 2:00 to 7:11 years (20 subjects in each age range). Subjects were selected and tested individually by the researcher. A comparative analysis of the subjects' performances with that observed in English-speaking children was made. RESULTS: the results indicated that Brazilian and American subjects presented similar performances until the age of 4:11 years. Portuguese-speaking children of 5, 6 and 7 years presented higher scores on the receptive subtest of Form A when compared to their American pairs. Portuguese-speaking subjects of 5 and 6 years presented lower scores on the expressive subtest of Form B when compared to their American pairs. CONCLUSION: the translation of diagnostic instruments that are already available in other languages can lessen the lack of such resources in countries where these instruments are not yet available. Furthermore, this practice can allow cross cultural studies which are relevant for researches on child language and language disabilities. PMID- 18408859 TI - [Performance of normal young adults in two temporal resolution tests]. AB - BACKGROUND: temporal auditory processing is defined as the perception of sound or of sound alteration within a restricted time interval and is considered a fundamental ability for the auditory perception of verbal and non verbal sounds, for the perception of music, rhythm, periodicity and in the discrimination of pitch, duration and of phonemes. AIM: to compare the performance of normal Brazilian adults in two temporal resolution tests: the Gaps-in-Noise Test (GIN) and the Random Gap Detection Test (RGDT), and to analyze potential differences of performance in these two tests. METHOD: twenty-five college students with normal hearing (11 males and 14 females) and no history of educational, neurological and/or language problems, underwent the GIN and RGDT at 40dB SL. RESULTS: statistically significant gender effects for both tests were found, with female participants showing poorer performance on both temporal processing tests. In addition, a comparative analysis of the results obtained in the GIN and RGDT revealed significant differences in the threshold measures derived for these two tests. In general, significantly better gap detection thresholds were observed for both male and female participants on the GIN test when compared to the results obtained for the RGDT. CONCLUSION: male participants presented better performances on both RGDT and GIN, when compared to the females. There were no differences in performance between right and left ears on the GIN test. Participants of the present investigation, males and females, performed better on the GIN when compared to the RGDT. The GIN presented advantages over the RGDT, not only in terms of clinical validity and sensibility, but also in terms of application and scoring. PMID- 18408860 TI - [Development skills in children with congenital hypothyroidism: focus on communication]. AB - BACKGROUND: congenital hypothyroidism may cause alterations in the child's global development. AIM: to outline the development profile in children with congenital hypothyroidism, focusing on communication, and to verify the influence of clinical history on the outlined profile. METHOD: 35 children, with ages between 2 to 36 months, with congenital hypothyroidism detected by neonatal screening, and who were in treatment for at least one month using hormonal replacement were assessed using the Early Language Milestone Scale (ELM) and the Portage Operation Inventory (POI). The clinical history was obtained in an interview with the family and from the analysis of medical records. RESULTS: in the ELM, eleven children presented a poor performance in the expressive auditory function, two in the visual function and one in the receptive auditory function. In the POI, seven children presented a poor performance in the language section, five in cognitive section, four in the motor and social sections and three in the self-care section. There was no correlation between the results obtained in the assessments and the clinical history. CONCLUSION: most of the children presented adequate performances in the evaluated skills. For the children with altered performance, larger deficits were observed in the language section, for the expressive aspects, and in the cognitive section. The influence of clinical history on the development profile was not confirmed. However, a tendency for an adequate performance was observed in those children who underwent neonatal screening, received an early diagnosis and treatment for the congenital hypothyroidism and who received higher doses of thyroxine at the beginning of treatment. The importance of a speech-language follow-up for communication development in this population is highlighted. PMID- 18408861 TI - [Phonological remediation program in students with developmental dyslexia]. AB - BACKGROUND: program for phonological remediation in developmental dyslexia. AIM: to verify the efficacy of a program for phonological remediation in students with developmental dyslexia. Specific goals of this study involved the comparison of the linguistic-cognitive performance of students with developmental dyslexia with that of students considered good readers; to compare the results obtained in pre and post-testing situations of students with dyslexia who were and were not submitted to the program; and to compare the results obtained with the phonological remediation program in students with developmental dyslexia to those obtained in good readers. METHOD: participants of this study were 24 students who were divided as follows: Group I (GI) was divided in two other groups - GIe with 6 students with developmental dyslexia who were submitted to the program; and GIc with 6 students with developmental dyslexia who were not submitted to the program; Group II (GII) was also divided in two other groups - GIIe with 6 good readers who were submitted to the program, and GIIc with 6 good readers who were not submitted to the program. The phonological remediation program (Gonzalez & Rosquete, 2002) was developed in three stages: pre-testing, training and post testing. RESULTS: results indicate that GI presented a lower performance in phonological skills, reading and writing when compared to GII in the pre-testing situation. However, GIe presented a similar performance to that of GII in the post-testing situation, indicating the effectiveness of the phonological remediation program in students with developmental dyslexia. CONCLUSION: this study made evident the effectiveness of the phonological remediation program in students with developmental dyslexia. PMID- 18408862 TI - [The use of verbal communicative abilities to increase the mean length of utterance in high-functioning autism and Asperger Syndrome]. AB - BACKGROUND: high-functioning autism (HFA) and Asperger Syndrome (AS) are pervasive developmental disorders that present alterations in the communicative and social skills. AIM: to use verbal communicative skill strategies (VCS) to increase the mean length of utterance (MLU) produced by individuals with HFA and AS. METHOD: participants were three 12-year-old males with HFA or AS. Data was collected from videotape recordings of structured verbal interaction sessions with the researcher and each participant during eight months. In order to verify the effects of the intervention, a multiple baseline research design across the participants was used, being composed by two phases: baseline (BL) and intervention (I). In the BL phase, spontaneous interaction situations occurred between the researcher and each participant. In the first phase of I, the intervention sessions occurred twice a week and gradually worked their way to once a week, but only after each participant had reached the goal of increasing the MLU. The number of sessions decreased gradually to avoid any drop in performance. The strategies which were used were divided in blocks of activities: spontaneous conversation; activities involving specific language difficulties; games with rules; story/report telling; and meta-linguistic activities. RESULTS: the application of the proposed strategies for verbal communicative abilities achieved the purpose of increasing the MLU of all three participants. CONCLUSION: suggestions are made for further researches that investigate the maintenance of the results in other environments and during the interaction with different communication partners. PMID- 18408863 TI - [Behavioral auditory processing evaluation in individuals with stuttering]. AB - BACKGROUND: the behavioral auditory processing (AP) evaluation allows the investigation of neuroaudiological processes involved in speech fluency processing. AIM: the purpose of this study was to describe the results obtained in the AP evaluation in stutterers, comparing the type of AP disorder with the severity of stuttering. METHOD: 56 subjects, 49 male and 7 female, ranging in age from 4 to 34 years, were referred from the speech-language clinic of UNIFESP to the AP evaluation. All patients were submitted to the following evaluations: audiological, speech and language. Disfluency was classified according to the protocol proposed by Riley (1994) which includes the following stuttering severity levels: very mild, mild, moderate, severe and very severe. Behavioral AP tests were selected and analyzed according to the patient's age and to the proposal of Pereira & Schochat (1997). RESULTS: subjects with ages between 4 to 7 years and between 12 to 34 years presented mostly mild stuttering, subjects between 8 to 11 years presented mostly moderate stuttering. From the total of 56 individuals who were evaluated, 92.85% presented AP disorders. The most common auditory processing disorders were supra-segmental and decoding. No statistical differences were found considering the results of the AP evaluation and the severity of stuttering for none of the age groups. CONCLUSION: the AP evaluation indicated deficits for most of the participants of all age groups, however no correlations were observed with the severity of stuttering. PMID- 18408864 TI - [Study of the hearing of rock and roll musicians]. AB - BACKGROUND: rock and roll has as one of its main characteristics the excessive sound pressure levels. Several studies have demonstrated that the sound levels of rock concerts can range from 100 to 115dB (A), with peak levels of 150dB (A). AIM: to study the hearing of rock and roll musicians, analyzing the results of the audiological evaluation and verifying the influence of time of exposure to amplified music. METHOD: a questionnaire was answered by 23 rock and roll musicians (46 ears) who were also evaluated by means of pure tone audiometry, immitance audiometry and transient/distortion product evoked otoacoustic emissions (OAET and OAEPD). RESULTS: regarding the time of exposure to music, values close to the limit of acceptance (tending to be significants) were found in the frequencies of 0.5 and 6kHz, in the pure tone audiometry. A statistically significant difference was also found in the OAET test in the frequency of 2kHz and also in the frequencies of 0.75, 1, 4 and 6kHz in the OAEPD test. CONCLUSION: the results indicate that although hearing loss was not found in the studied population, alteration in the register of the OAE already exists, suggesting alteration of the cochlear function. Regarding time of exposure, the results indicate that musicians with more than 10 years of practice present statistically significant differences when compared to those with less time of exposure. PMID- 18408865 TI - [Characterization of language disorders in children with lead poisoning]. AB - BACKGROUND: lead poisoning can have a negative impact on the neuropsychological functions, including language, due to the damage it causes to the development of the Central Nervous System. AIM: to verify the occurrence of language disorders in children who suffered from led poisoning and to verify the correlation between the lead concentration level in the blood and the language disorders presented by the children. METHOD: language evaluation of 20 preschoolers, with lead concentration level in the blood above 10 microg/dl. RESULTS: 13 children presented language impairment involving only phonology or more than one language subsystem. The statistical analysis indicated that no correlation exists between the severity of the language impairment and the concentration levels of lead. CONCLUSION: the number of children with language impairment indicates lead poisoning as a risk factor for the present alterations, even though other risk factors for language disorders were found and the absence of correlation between the investigated variables. PMID- 18408866 TI - [Changes in behavior associated to the use of voice after a speech therapy intervention with professionals of child day care centers]. AB - BACKGROUND: preventive programs have been used with teachers with the purpose of lowering the number of risk factors for voice disorders. However, few studies have focused on the effectiveness of these programs. AIM: to assess changes in behaviors that are considered, according to the specific literature, as having a negative impact on the voice, during a speech therapy intervention program offered to educators. METHOD: an experimental study where a theoretical-practical program involving the professional use of voice was developed with 26 educators from two day care centers of Sao Paulo. This program involved five monthly meetings, working up to a total of twelve hours. During the program, topics concerning negative behaviors that affect the voice were discussed. On four of the meetings the educators filled in a protocol, making it possible to analyze the frequency on which they presented those specific behaviors. Later on, scores were given to the answers on the protocol in order to compare the findings during the program. For this comparison the statistical test of Wilcoxon and the linear tendency chi-square test were used. RESULTS: the gradual decrease in the use of voice out of work was observed, as well as in speaking with a very low or high pitch voice and in eating excessively before sleeping. There was an increase in the occurrence of coughing in the third application of the protocol. As for the overall monthly averages, statistically significant differences were not found when comparing the four applications of the protocol. The overall average score was of 11.75, which was considered moderately abusive. CONCLUSION: the changes that were observed during the program were interesting, but very restrict which makes one wonder about the real effects of this kind of practice and how much broader changes rely on small individual changes. Actions that involve information about vocal well-being exclusively should be revised. PMID- 18408867 TI - [Speech and non-speech activities in stuttering: a preliminary study]. AB - BACKGROUND: stuttering. AIM: to compare muscle activation in fluent and stuttering individuals during speech and non-speech tasks. METHOD: six adults divided in two groups: G1 - three fluent individuals; G2 - three stuttering individuals. Muscle activity (surface electromyography) was captured by disposable electrodes fixed in four regions. Testing situations: muscle rest tension, speech reaction time, non-verbal activity, verbal activity. RESULTS: There was no significant statistical difference between the groups for the rest tension; G2 present longer speech reaction times; G2 presented muscle activity during the non-verbal task similar to that observed during rest; Muscle activity of G1 and G2 during the verbal task demonstrated to be similar. CONCLUSION: these results suggests that for G2 there is a poor control of timing for the coordination of motor processes. PMID- 18408868 TI - Influence of cellulose polymers on the semipermanent dyestuff process for yak hair: an analytical investigation. AB - The aim of this study was to investigate the effect of some cellulose polymers mixed in a semipermanent dyestuff on the dye uptake of yak hair fiber and color fading after repeated washing cycles. Two different classes of commercial polymers were tested: non-ionic and cationic. Formulations based on a mixture of HC and basic dyes, with different molecular sizes, were employed as representative dyestuffs. UV-Vis spectroscopy and colorimetric measurements were used to analyze the extracted dyes from the yak hair cuticle and cortex. The results obtained indicate that the presence of cationic polymers in the dye bath improves both the quality of the dyeing process and the anti-fading effect during the first washing cycles. PMID- 18408869 TI - A dual mechanism of 4-hydroxy-5-methyl-3[2H]-furanone inhibiting cellular melanogenesis. AB - In previous studies, 4-hydroxy-5-methyl-3[2H]-furanone (HMF) was shown to have potent antioxidative and antimelanogenic effects, suggesting its potential use as a depigmenting agent. The present study investigated its mechanism of action on murine melanoma B16F10 cells stimulated by theophylline, an activator of the cyclic AMP/protein kinase A signaling leading to tyrosinase gene expression. When the cells were stimulated with theophylline, there were dose-dependent increases in cellular tyrosinase protein content and melanin formation, as expected. HMF inhibited the theophylline-stimulated melanin formation as effectively as arbutin, one of the most widely used depigmenting agents in cosmetics. HMF appeared to reduce tyrosinase mRNA and protein content in the cells stimulated by theophylline, indicating it inhibited tyrosinase gene expression. HMF also effectively inhibited tyrosinase-catalyzed melanin formation from dihydroxyphenylalanine in the cells as well as in vitro. Therefore, the antimelanogenic effects of HMF were best explained by a dual mechanism inhibiting tyrosinase gene expression and the enzyme activity of pre-existing tyrosinase. PMID- 18408870 TI - Why women use makeup: implication of psychological traits in makeup functions. AB - Makeup acts and stimulates three of our senses: touch (which encompasses all sensations from the body surface), smell (fragrance), and sight (the process of becoming and looking beautiful). The positive stimulation of these senses by makeup can induce sensory as well as psychological pleasure. In order to understand the relationship of women to their makeup, we interviewed different groups of women on their quality of life and makeup habits. Then, through four standard well-validated psychometric self-questionnaires, we examined the possible relation between the need to make up oneself and specific psychological features. Our first results clearly showed that makeup could support two opposite "up" functions, i.e., "camouflage" vs "seduction." Concerning their psychological profiles, results showed that women of the functional class "camouflage" are more anxious, defensive, and emotionally unstable compared to those of the functional class "seduction," who appear to be more sociable, assertive, and extroverted. Further analyses revealed a division of the two classes into subclasses of volunteers with opposed personality and psychological profiles. This new classification allowed us to define more precisely the relations existing within the subjective experience of women during the makeup process. In conclusion, our study revealed that beyond the simple application of colorful products on the face, makeup has two major functional implications depending on specific psychological profiles of women. PMID- 18408871 TI - Enhanced depigmenting effects of N-glycosylation inhibitors delivered by pH sensitive liposomes into HM3KO melanoma cells. AB - Delivery activity of pH-sensitive 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE):cholesteryl hemisuccinate (CHEMS) liposomes was assessed as an in vitro intracellular carrier system to increase the bioavailability of depigmentation actives. N-glycosylation inhibitors have a glycosylation-inhibiting effect, which is useful for the skin depigmentation that operates by interfering with the maturation of tyrosinase. However, an N-glycosylation inhibitor does not easily pass through skin or even cellular membranes due to its water-soluble property. Therefore, it should be transported to target cells by an efficient delivery carrier to reduce the glycosylated tyrosinase. Glycosylation-inhibiting and depigmentation effects of N-butyldeoxynojirimycine (NB-DNJ) and 1 deoxynojirimycine (DNJ)-loaded liposomes were evaluated using Western blotting and measurement of synthesized melanin. Interestingly, it was found that the pH sensitive liposomes increased the glycosylation-inhibiting and thus, pigment lightening effects of N-glycosylation inhibitors in vitro. In addition, cargo materials loaded in pH-sensitive liposomes were found to be much more efficiently delivered into the cytoplasm, as observed in fluorescent-activated cell sorting (FACS) and confocal laser-scanning microscopic (CLSM) analysis. These results indicate that pH-sensitive DOPE:CHEMS liposomes have a strong potential as a carrier system to promote delivery efficiency and to enhance the biological effects of water-soluble actives for applications in cosmetics, personal care products, and pharmaceutics. PMID- 18408872 TI - Comparison of hair shaft damage after UVA and UVB irradiation. AB - Sunlight, especially ultraviolet (UV) light-induced hair damage is difficult to avoid during daily life. Concerns about the effects of ultraviolet light on hair are emerging recently. These photochemical changes mainly come from damage to hair proteins and melanins. In this study, we performed experiments to find the patterns of morphological and biochemical changes in UV-light-induced damage to hair by scanning and transmission electron microscopy and hair protein analysis. In our results, morphological damage is significant in UVB-irradiated hairs, while biochemical changes are greater in UVA-irradiated hairs. PMID- 18408878 TI - Toxic Effects in Siganus oramin by dietary exposure to 4-tert-octylphenol. AB - The integrated toxicities of 4-tert-octylphenol (t-OP) on Siganus oramin were investigated by dietary administration at doses of 5, 25 and 125 mg/kg body weight over 28 days. Significant increase was observed in the activity of hepatic glutathione S-transferase at 125 mg/kg on both day 14 and 28 in males, and at all doses on day 28 in females, and in hepatosomatic index at 25 mg/kg on day 14 in both sexes. Plasma levels of testosterone and cortisol decreased significantly at all doses on day 28. Histopathologic changes in liver, spleen, intestine and testis deteriorated with increasing doses and duration. The results suggest that S. oramin is sensitive to t-OP, and the above endpoints may be potential biomarkers for evaluating toxicities of environmental pollutants such as t-OP. PMID- 18408879 TI - Association between quantitative measures of skin color and plasma 25 hydroxyvitamin D. AB - We examined the relationship between vitamin D and skin color measured by reflectance colorimetry at an exposed and un-exposed site in 321 people. Exposed but not unexposed skin color was associated with better vitamin D status. Sun exposure was more important than natural skin color in determining vitamin D status in our population. INTRODUCTION: Vitamin D is obtained through UV synthesis in the skin where melanin limits its synthesis. Ethnicity is often used as a proxy for skin color, but skin color varies considerably. The relation between quantitative measures of skin color and plasma 25-hydroxyvitamin D (25OHD) concentration has not been well described. METHODS: The aim of this study was to determine the association between constitutive (natural) and sun-induced skin color and 25OHD in a group of Pacific People (n = 87) and Europeans (n = 255) living in NZ (46 degrees S) in summer. Plasma 25OHD was determined and sun induced (outer fore-arm) and constitutive (upper inner-arm) measured by reflectance colorimetry. RESULTS: Mean (SD) 25OHD was significantly higher in Europeans than Pacific People, 88 (31) nmol/L vs. 75 (34) nmol/L, respectively. Based on constitutive skin color, 35% of participants were very light, 45% light, 16% intermediate, 4% tanned, and 0% brown or dark. Skin color at the forearm but not constitutive skin color was a significant predictor of 25OHD. Each 10 degrees lower skin color value at the forearm (more tanning) was associated with a 5 nmol/L higher 25OHD (P < 0.001). CONCLUSIONS: Tanning but not natural skin color was an important determinant of 25OHD. Further study is needed in a population with a higher proportion of darker skin people. PMID- 18408880 TI - Self-reported sleep in systemic lupus erythematosus. AB - The objectives of the study were to assess sleep disturbances in systemic lupus erythematosus (SLE) and to compare these with a working sample and a treatment seeking sample reporting insomnia. The primary sample was 172 people with SLE. This sample represented 32% of all members of two lupus support association. Two comparison samples were used: 223 adults who expressed interest in taking part in a psychological treatment for sleep problems and 456 Australian adults who were working at a large organization. All individuals completed the Pittsburgh Sleep Quality Index (PSQI; 6). Data derived from the PSQI included total sleep time, sleep onset latency, wake after sleep onset, sleep efficiency, as well as the global and seven component scores. The SLE sample reported significantly worse sleep on all parameters than the working sample, but significantly better sleep than the sample of those seeking treatment for sleep disorders, except for sleep onset latency. The percentages scoring >5 on the PSQI global score was 80.5% for SLE, 91.5% for those seeking treatment for sleep disorders, and 28.5% for the working sample. PSQI component scores for the SLE group more closely resembled those of the treatment-seeking group. Self-reported sleep in this sample of people with SLE was significantly better on most parameters than that of a group seeking treatment for sleep disorders. However, the values obtained tended to be worse than previous reports and indicated less than optimal sleep. However, the low response rate of the sample was of concern and may indicate that the sample was biased. The present results suggest that sleep disturbance is common in those with SLE and deserves more attention in a more representative sample. PMID- 18408881 TI - Increased MS relapse rate during assisted reproduction technique. PMID- 18408882 TI - Time to insulin in type-2 diabetes: high hurdles or Santiago way? AB - The prevalence of obesity has been increasing dramatically in the last decades; the major metabolic complication of obesity is insulin resistance and type-2 diabetes because there are pathogenetic mechanisms linking obesity and type-2 diabetes. Diabetes is also rapidly increasing worldwide; such a description of the key stages in the evolution of type-2 diabetes may be of great interest for implementing antidiabetes treatment. In recent times, type-2 diabetes therapy has been based on drugs, which improve insulin sensibility or stimulate insulin secretion or slow down glucose absorption. Recently, an ADA and EASD consensus has been released to develop a common approach for the management of hyperglycaemia in adults. The development of new classes of blood-glucose lowering medications to supplement the older therapies, such as lifestyle directed interventions, insulin, sulfonylureas, and metformin, has increased the different possible options for the treatment of type-2 diabetes. Therapeutic approaches aiming to potentiate the biological effects of incretins include degradation-resistant GLP-1 receptor agonists (incretin mimetics), and inhibitors of dipeptidyl peptidase-IV (DPP-IV) activity (incretin enhancers) will be very useful to slow down type-2 diabetes progression. Weight-loss interventions, such as a hypocaloric diet and physical exercise, in addition to agents such as orlistat, sibutramine and cannabinoid receptor antagonists, may have favourable effects upon fat storage, nutrient metabolism and ultimately glucose tolerance or type-2 diabetes. When the therapeutic target is not achieved, insulin with metformin could be suggested, but is this approach the ideal one for all patients? Perhaps it is possible to delay the initiation of insulin therapy, therefore, the actual and future therapeutical options are considered in the present review. PMID- 18408883 TI - Cellular prion protein prevents brain damage after encephalomyocarditis virus infection in mice. AB - Cellular prion protein (PrP(C)), a cell-surface glycoprotein normally associated with neurons, is also expressed in other cell types such as glia and lymphocytes. To further elucidate these roles of PrP(C), wild-type prion protein gene (Prnp(+/+)) mice and Prnp-deficient (Prnp(-/-)) mice were infected with encephalomyocarditis virus B variant (EMCV-B) via an intracranial route. EMCV-B causes encephalitis and apoptotic cell death in vivo. Histopathological studies revealed that Prnp(+/+) mice infected with 600 pfu of EMCV-B showed more severe infiltration of inflammatory cells, accompanied by higher activation of microglia cells around the hippocampus, than Prnp(-/-) mice; viz., no differences in the brain virus titer between these two lines of mice. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP, nick end-labeling (TUNEL) staining of the brain specimens revealed that the CA1 hippocampal pyramidal cells showed a larger number of apoptotic neurons in Prnp(-/-) than Prnp(+/+) mice. Based on all these findings, PrP(C) may play certain roles in the induction of inflammation and inhibition of apoptosis in vivo. PMID- 18408884 TI - Activation-induced cellular accumulation of histamine in immature but not mature murine mast cells. AB - Mast cell activation involves the rapid release of inflammatory mediators, including histamine, from intracellular granules. The cells are capable of regranulation and multiple rounds of activation. The goal of this study was to determine if there are changes in the content of pre-formed mast cell mediators after a round of activation. After 24 h, the histamine content of bone marrow derived mast cells (BMMC), but not that of peritoneal mast cells, exceeded the amount in resting cells. Accumulation of histamine in BMMC peaked at 72 h of activation, and returned toward preactivation levels by 96 h. The increase in histamine content was accompanied by an increase in the gene expression of histidine decarboxylase. No increases in beta hexosaminidase or murine mast cell protease-6 were observed. These findings indicate that BMMC respond to activation by increasing total cell-associated histamine content. This increase may be important to the response of these cells upon subsequent exposure to antigens. PMID- 18408885 TI - Large-scale production of functional membrane proteins. AB - The preparation of sufficient amounts of high-quality samples is still the major bottleneck for the characterization of membrane proteins by in vitro approaches. The hydrophobic nature, the requirement for complicated transport and modification pathways, and the often observed negative effects on membrane properties are intrinsic features of membrane proteins that frequently cause significant problems in overexpression studies. Establishing efficient protocols for the production of functionally folded membrane proteins is therefore a challenging task, and numerous specific characteristics have to be considered. In addition, a variety of expression systems have been developed, and choice of appropriate techniques could strongly depend on the desired target membrane proteins as well as on their intended applications. The production of membrane proteins is a highly dynamic field and new or modified approaches are frequently emerging. The review will give an overview of currently established processes for the production of functionally folded membrane proteins. PMID- 18408886 TI - Pharmacological manipulation of L-carnitine transport into L6 cells with stable overexpression of human OCTN2. AB - The high-affinity Na+-dependent carnitine transporter OCTN2 (SLC22A5) has a high renal expression and reabsorbs most filtered carnitine. To gain more insight into substrate specificity of OCTN2, we overexpressed hOCTN2 in L6 cells and characterized the structural requirements of substances acting as human OCTN2 (hOCTN2) inhibitors. A 1905-bp fragment containing the hOCTN2 complete coding sequence was introduced into the pWpiresGFP vector, and L6 cells were stably transduced using a lentiviral system. The transduced L6 cells revealed increased expression of hOCTN2 on the mRNA, protein and functional levels. Structural requirements for hOCTN2 inhibition were predicted in silico and investigated in vitro. Essential structural requirements for OCTN2 inhibition include a constantly positively charged nitrogen atom and a carboxyl, nitrile or ester group connected by a 2-4-atom linker. Our cell system is suitable for studying in vitro interactions with OCTN2, which can subsequently be investigated in vivo. PMID- 18408887 TI - Mechanisms of voltage-gated ion channel regulation: from gene expression to localization. AB - The ion channel milieu present in a neuron in large part determines the inherent excitability of a given cell and is responsible for the translation of sensory transduction and synaptic input to axonal output. Intrinsic excitability is a dynamic process subject to multiple levels of regulation from channel gene expression to post-translational modifications that influence channel activity. The goal of this review is to provide an overview of some of the mechanisms by which channels can be modified in order to influence neuronal output. We focus on four levels of regulation: channel gene transcription, alternative splicing of channel transcripts, post-translational modifications that alter channel kinetics (phosphorylation), and subcellular localization and trafficking of channel proteins. PMID- 18408888 TI - The post-transcriptional steps of eukaryotic ribosome biogenesis. AB - One of the most important tasks of any cell is to synthesize ribosomes. In eukaryotes, this process occurs sequentially in the nucleolus, the nucleoplasm and the cytoplasm. It involves the transcription and processing of pre-ribosomal RNAs, their proper folding and assembly with ribosomal proteins and the transport of the resulting pre-ribosomal particles to the cytoplasm where final maturation events occur. In addition to the protein and RNA constituents of the mature cytoplasmic ribosomes, this intricate process requires the intervention of numerous protein and small RNA trans-acting factors. These transiently interact with pre-ribosomal particles at various stages of their maturation. Most of the constituents of pre-ribosomal particles have probably now been identified and research in the field is starting to unravel the timing of their intervention and their precise mode of action. Moreover, quality control mechanisms are being discovered that monitor ribosome synthesis and degrade the RNA components of defective pre-ribosomal particles. PMID- 18408889 TI - The leucine-rich repeat structure. AB - The leucine-rich repeat is a widespread structural motif of 20-30 amino acids with a characteristic repetitive sequence pattern rich in leucines. Leucine-rich repeat domains are built from tandems of two or more repeats and form curved solenoid structures that are particularly suitable for protein-protein interactions. Thousands of protein sequences containing leucine-rich repeats have been identified by automatic annotation methods. Three-dimensional structures of leucine-rich repeat domains determined to date reveal a degree of structural variability that translates into the considerable functional versatility of this protein superfamily. As the essential structural principles become well established, the leucine-rich repeat architecture is emerging as an attractive framework for structural prediction and protein engineering. This review presents an update of the current understanding of leucine-rich repeat structure at the primary, secondary, tertiary and quaternary levels and discusses specific examples from recently determined three-dimensional structures. PMID- 18408890 TI - Substrate specificity of bacterial DD-peptidases (penicillin-binding proteins). AB - The DD-peptidase enzymes (penicillin-binding proteins) catalyze the final transpeptidation reaction of bacterial cell wall (peptidoglycan) biosynthesis. Although there is now much structural information available about these enzymes, studies of their activity as enzymes lag. It is now established that representatives of two low-molecular-mass classes of DD-peptidases recognize elements of peptidoglycan structure and rapidly react with substrates and inhibitors incorporating these elements. No members of other DD-peptidase classes, including the high-molecular-mass enzymes, essential for bacterial growth, appear to interact strongly with any particular elements of peptidoglycan structure. Rational design of inhibitors for these enzymes is therefore challenging. PMID- 18408891 TI - Pituitary metastasis from medullary carcinoma of thyroid: case report and review of literature. AB - Pituitary is a rare site for metastases from thyroid cancer. Most reported cases have been of papillary and follicular carcinoma. Metastases from medullary thyroid carcinoma have not been reported. We report a case of intrapituitary metastasis from medullary carcinoma thyroid in a 38-year-old male, who had been operated for pituitary adenoma 5 years earlier. At the time of presentation in Nov 2006, he had visual field defects and a painless thyroid nodule. Further evaluation revealed medullary carcinoma thyroid, cervical and mediastinal lymphadenopathy, elevated serum calcitonin levels, and lobulated pituitary tumor. After surgical excision of thyroid and lymph node clearance, he underwent craniotomy and subfrontal excision of pituitary tumor. All the tumors were of identical histology, i.e., medullary carcinoma thyroid. Pituitary tumor was positive for calcitonin. PMID- 18408892 TI - Multifocal glioblastoma multiforme with synchronous spontaneous hemorrhage: case report. AB - We report a 65 year old man who presented with left hemiparesis, and was found to have multiple, discrete, peripherally enhancing, hemorrhagic intra-axial masses in the right hemisphere of the brain. Workup for malignancy elsewhere in the body was negative, and biopsy confirmed glioblastoma multiforme. The patient responded clinically to treatment with radiation therapy and temozolomide. We discuss the unusual aspects of this case and stress the importance of tissue diagnosis in managing suspected intracranial malignancies. PMID- 18408894 TI - Response of Pseudomonas putida F1 cultures to fluctuating toluene loads and operational failures in suspended growth bioreactors. AB - The response of Pseudomonas putida F1 to process fluctuations and operational failures during toluene biodegradation was evaluated in a chemostat suspended growth bioreactor. The ability of P. putida F1 to rapidly increase its specific toluene degradation capacity resulted in no significant variation in process removal efficiency when toluene load was increased from 188 to 341 g m(-3) h(-1). Likewise, bacterial activity rapidly reached steady state performance (in less than 1.5 h after the restoration of steady state operational conditions) following an 8 h process shutdown, or after episodes of toluene or mineral nutrients deprivation. Process performance was however highly sensitive to pH, as pH levels below 4.5 dramatically inhibited bacterial activity, decreasing severely process robustness and inducing a cycle of periodic process collapses and recoveries. This pH mediated deterioration of bacterial activity was confirmed by further respirometric tests, which revealed a 50-60% reduction in the O(2) consumption rate during the degradation of both toluene and 3-methyl catechol when pH decreased from 5.05 to 4.55. Finally, process robustness was quantified according to methods previously described in literature. PMID- 18408893 TI - Chemoprevention of colon carcinogenesis by oleanolic acid and its analog in male F344 rats and modulation of COX-2 and apoptosis in human colon HT-29 cancer cells. AB - PURPOSE: To assess the chemopreventive effect of oleanolic acid (ONA) and its synthetic analog 18alpha-olean-12-ene-3beta-23,28-triol (OT) on azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) in F344 rats and understand anti inflammatory properties and apoptosis effects in HT29 colon cancer cells and Raw 264.7 macrophage cell lines. METHODS: Five week-old male F344 rats were fed a control diet or experimental diets containing two doses of ONA (750 and 1,500 ppm) and OT (250 and 500 ppm). After 1 week, all animals were s.c. injected with AOM (15 mg/kg body weight, once weekly for 2 weeks). At 14 weeks of age, all rats were killed and colons were evaluated for ACF. Cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) expressions and apoptosis were assessed in cell lines exposed to OT using western blots and 4',6-diamidino-2-phenylindole staining. RESULTS: Administration of ONA and OT inhibited mean colonic ACF and multi-crypt AC/foci in a dose dependent manner (p < 0.001-0.0001). OT blocked the COX-2 expression induced by phorbol 12-myristate 13-acetate in a dose-dependent manner and induced apoptosis in HT-29 cancer cells, and suppressed iNOS activation in RAW264.7 macrophages. CONCLUSIONS: ONA and OT possess chemopreventive activity against colon carcinogenesis in rat and OT inhibits the COX-2 and iNOS and induces apoptosis in cell lines. PMID- 18408895 TI - Investigation on bioavailability of some essential and toxic elements in medicinal herbs. AB - Trace and major elements were determined in medicinal herbs (Cynara scolymus, Matricaria chamomilla, Artemisia absinthium L., Achillea millefolium, and Inula britannica) as well as in rhizosphere soil samples. Based on the results obtained after microwave-acid-assisted digestion (nitric acid + hydrogen peroxide) and single-step extraction (ammonium acetate), the real and potential acidity and redox potential of the soils, uptake, mobility, and bioavailability of potassium, calcium, magnesium, iron, manganese, copper, zinc, nickel, chromium, lead, and cadmium are discussed. By calculating the bioconcentration factors and their deviation from the recommended values, elevated concentrations, were explained in terms of contamination and pollution. The concentrations measured in both plants and soil samples were below maximum allowable concentration ranges considered for the European Union. PMID- 18408896 TI - Evaluation of Sesbania grandiflora for antiurolithiatic and antioxidant properties. AB - In the indigenous system of medicine in India, the plant Sesbania grandiflora is claimed to be useful for various ailments, and one such use is for the treatment of renal calculi. The major purpose of this study is to investigate the potential of S. grandiflora in the treatment of renal calculi. The leaf juice of S. grandiflora was evaluated for median lethal dose, gross behavioral changes, antiurolithiatic and antioxidant activities. The antiurolithiatic activity was evaluated by a calculi-producing diet model, using gentamicin (subcutaneously) and 5% ammonium oxalate in rat feed to induce calcium oxalate-type stones. The parameters monitored in the present study are calcium and oxalate deposition in the kidney, kidney weights, urinary excretion of calcium and oxalate. The in vivo antioxidant parameters lipid peroxidation, glutathione reductase and catalase were monitored. The plant juice was also evaluated for scavenging of nitric oxide and 2-diphenyl-2-picryl hydrazyl free radicals. The leaf juice of S. grandiflora was safe orally and exhibited no gross behavioral changes except for an increase in urination. The leaf juice of S. grandiflora showed significant antiurolithiatic activity against calcium oxalate-type stones and also exhibited antioxidant properties. The results obtained in this study provide evidence for the efficacy of the leaf juice of S. grandiflora as antiurolithiatic agent. PMID- 18408897 TI - Relationships among iron, protein oxidation and lipid peroxidation levels in rats with alcohol-induced acute pancreatitis. AB - It has been previously shown that alcohol induces the damage of pancreatic parenchyma tissue, but the mechanism of this damage is still poorly understood. Assuming that oxygen radical damage may be the involved, we measured markers of oxidative damage in pancreatic tissue, blood serum, plasma, and whole blood of rats with early-stage alcohol-induced acute pancreatitis. Thirty-eight male Wistar rats were divided into three groups: the control group (group 1), the acute pancreatitis group 1 day (group 2), and 3 days (group 3) after the injection of ethyl alcohol into the common biliary duct, respectively. The levels of Fe in tissue and serum, whole blood viscosity, plasma viscosity, fibrinogen and homocysteine (Hcy) levels, erythrocyte and plasma malondialdehyde (MDA), and tissue and plasma protein carbonyl levels were found to be significantly higher in groups 2 and 3 than in group 1. However, the levels of reduced glutathione (GSH) in tissue and erythrocytes were significantly lower in groups 2 and 3 than in group 1. These results suggest that elevated Fe levels in serum and pancreatic tissue in rats with early-stage alcohol-induced acute pancreatitis is associated with various hemorheological changes and with oxidative damage of the pancreas. PMID- 18408898 TI - Effects of pre- and postnatal exposure to chromium picolinate or picolinic acid on neurological development in CD-1 mice. AB - Chromium picolinate, Cr(pic)3, a popular dietary supplement marketed as an aid in fat loss and lean muscle gain, has also been suggested as a therapy for women with gestational diabetes. The current study investigated the effects of maternal exposure to Cr(pic)3 and picolinic acid during gestation and lactation on neurological development of the offspring. Mated female CD-1 mice were fed diets from implantation through weaning that were either untreated or that contained Cr(pic)3 (200 mg kg(-1) day(-1)) or picolinic acid (174 mg kg(-1) day(-1)). A comprehensive battery of postnatal tests was administered, including a modified Fox battery, straight-channel swim, open-field activity, and odor-discrimination tests. Pups exposed to picolinic acid tended to weigh less than either control or Cr(pic)3-exposed pups, although the differences were not significant. Offspring of picolinic acid-treated dams also appeared to display impaired learning ability, diminished olfactory orientation ability, and decreased forelimb grip strength, although the differences among the treatment groups were not significant. The results indicate that there were no significant effects on the offspring with regard to neurological development from supplementation of the dams with either Cr(pic)3 or picolinic acid. PMID- 18408899 TI - Introduction to session 1A: feedstock genomics and development. PMID- 18408900 TI - Introduction to session 2: enzyme catalysis and engineering. PMID- 18408901 TI - [74th Annual Meeting of the German Cardiology Society, 27-29 March 2008, Mannheim, Germany. Abstract database]. PMID- 18408902 TI - Skeletal age assessment from elbow radiographs. Review of the literature. AB - Skeletal age is important to evaluate remaining growth. In 50% of normal children and adolescents, skeletal age does not differ from chronological age. During puberty, skeletal age is an important tool when performing a lower limb epiphysiodesis or when treating (conservatively or surgically) patients with spinal deformities. Skeletal age alone is not enough and should be assessed together with other clinical and radiological findings such as standing and sitting heights, Risser sign, Tanner stages and annual growth rate. Puberty starts at 11 years of skeletal age and ends at 13 years of skeletal age in girls; in boys, puberty starts two years later (13 years of skeletal age) and then ends at a skeletal age of 15. Most current clinical and radiographic markers do not help paediatric orthopaedic surgeons to clearly distinguish maturity levels prior to Risser I. Sauvegrain et al. developed a method to assess skeletal age by using elbow radiographs (AP and lateral projections). Between 11 and 13 years of skeletal age in girls and between 13 and 15 years of skeletal age in boys, the olecranon apophysis is characterised by a clear morphological development. This method is a reliable tool to assess skeletal age during puberty because significant morphological changes in the elbow happen every six months. PMID- 18408903 TI - Inhibition of monoamine oxidase-B by the polyphenolic compound, curcumin and its metabolite tetrahydrocurcumin, in a model of Parkinson's disease induced by MPTP neurodegeneration in mice. AB - We investigated the effects of the polyphenolic compound curcumin and its metabolite tetrahydrocurcumin (ThC), in the model of Parkinson's disease induced in mice by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). In this model depletion of dopamine(DA) and DOPAC (3,4-dihydroxy phenyl acetic acid)) occurs with increased monoamine oxidase (MAO-B) activity. We used HPLC with electrochemical detection to measure DA and DOPAC respectively while MAO-B was assayed by spectroflourimetry using the conversion of the fluorogenic substrate, kyuramine. Systemic administration of curcumin (80 mg/kg i. p.) and tetrahydrocurcumin (60 mg/kg i. p.) significantly reversed the MPTP-induced depletion of DA and DOPAC. The MAO-B activity was also significantly inhibited by these compounds. The results showed that curcumin and tetrahydrocurcumin reversed the MPTP induced depletion of DA and DOPAC which may in part be due to inhibition of MAO-B activity. In conclusion, both curcumin and its metabolite ThC exert neuroprotection against MPTP induced neurotoxicity. PMID- 18408906 TI - [Impending Coumarin-necrosis in a patient with heterozygous protein C deficiency type I]. AB - Painful pink or magenta colored skin lesions characterized by a clear line of demarcation between the affected area and surrounding tissue appearing under therapy with coumarins may be a sign for evolving coumarin-necrosis. Immediate treatment with a protein C preparation in patients with protein C deficiency can prevent necrosis. PMID- 18408907 TI - [Simple and complex post-traumatic stress disorders. Diagnostic and therapeutic approaches]. AB - The prevalence rates of post-traumatic stress disorders are high in the general population (5-10%). The main diagnostic criteria include the experience of an event of extraordinary threat and the persistence of specific symptoms such as intrusion, avoidance, and physiological hyperarousal. Long-lasting traumata may lead to the development of complex syndromes or irreversible personality alterations. Chronic manifestations, psychiatric comorbidities, and psychosocial deficits are special risks in the course of post-traumatic stress disorders. Severe traumata are associated with complex neurobiological changes. Psychotherapeutic approaches are established as a three-stage model: stabilisation, trauma reorientation, and psychosocial reintegration. The additional use of psychotropic drugs should be oriented to the specific symptomatology. PMID- 18408908 TI - [Unclear parietal swelling synchronous with screaming]. PMID- 18408909 TI - [Urological update seminar "UroUpdate". Comprehensive 2-day course in continuing education]. PMID- 18408910 TI - [New guidelines for urinary stone treatment. Controversy or development?]. AB - The new guidelines of the European (EAU), American (AUA), German, and Austrian associations of urology are based on thorough and consistent research and analysis of the published literature. However, the methodology is very diverse. In the case of the EAU-AUA guidelines on ureteral stones, the evidence was generated by a laborious meta-analysis of the entire available published literature on the subject. These guidelines represent the 1A level of evidence. The traditional European guidelines evaluate each statement separately and assign a level of evidence to each. The following aspects are new: the even greater effort to formulate guidelines accompanied by the initiation of international cooperative projects, clarity of presentation and evaluation of the individual statements (including levels of evidence), information on medication-based treatment to facilitate spontaneous passage of ureteral stones as well as stone fragments after extracorporeal shock wave lithotripsy (ESWL), a shift in the value of ESWL and ureterorenoscopy for ureteral stones, retrograde intrarenal surgery for small kidney stones refractory to ESWL or in settings unsuitable for ESWL, the use of laparoscopy in those rare cases that represent an indication for open surgery, and the additional value given to metaphylaxis in diagnosis and therapy. All of the new guidelines take these developments into account, although they differ slightly in the importance they assign to the individual items. The modifications represent the developments secondary to electronic data processing and preparation on the one hand and the rapid development of medical instruments on the other. Although ESWL is still the most important procedure for treating urinary stones, advances in flexible endoscopes, intracorporeal lithotripsy, and extraction instruments have led to a shift in the range of indications. These developments are fully accommodated in the new guidelines. PMID- 18408911 TI - [Early detection of prostate cancer in Germany. A study of a representative random sample of the population]. AB - The goals of our study were to assess the prevalence of prostate cancer screening (PCS) among German men in terms of regularity of use and to analyze predictors of PCS use. A representative sample of 10,659 men aged 45-70 (mean=55.2) were surveyed to self-assess regularity of DRE and PSA tests; sociodemographic variables (age, income, education, marital status and health insurance status), family history of cancer, physician recommendation and medical checkups were assessed as well.Two thirds of the sample report ever undergoing a DRE; 48% had ever had a PSA test. In contrast, the rates of men who undergo PCS regularly are lower (44% DRE, 33% PSA). PCS increases with age; socioeconomic variables (such as education or income), however, are less important predictors. Family history of cancer is associated with PCS use, but not with regularity of use. The most important predictors are having medical checkups and physician recommendation. PMID- 18408912 TI - [Combined flexible and semirigid ureteroscopy with laser lithotripsy. Alternative to percutaneous nephrolithotomy of complex EWSL refractory nephrolithiasis]. AB - BACKGROUND: Flexible ureteroscopic laser lithotripsy of intrarenal calculi is technically demanding and challenging due to potential dislocation from the calyx and damage to the tissue or the fiber optic during in situ laser lithotripsy. MATERIAL AND METHODS: In 12 consecutive patients ESWL resistant renal stones were collected with the flexible ureteroscope using the nitinol basket and repositioned into the renal pelvis. After removing the flexible ureteroscope from the ureter the stones were treated through an additionally inserted semirigid ureteroscope with frequency-doubled double-pulse Neodym:YAG (FREDDY) laser under direct vision. RESULTS: Operating time was on average 110 min. During the complication-free 3 month follow-up, 8 patients had complete success, 3 partial success, 1 patient had residual fragments>3 mm and 88% of lower pole calyces were stone-free. CONCLUSIONS: As an alternative to percutaneous nephrolithotomy, repositioning of renal stones to the renal pelvis with the flexible instrument allows in selected cases safe laser fragmentation through a semirigid instrument. Access or exposition-related problems can be solved and repair costs of the instruments will be minimized. PMID- 18408914 TI - Doppler monitoring of renal hemodynamics: why the best is yet to come. PMID- 18408913 TI - A potential role for skeletal muscle caveolin-1 as an insulin sensitivity modulator in ageing-dependent non-obese type 2 diabetes: studies in a new mouse model. AB - AIMS/HYPOTHESIS: Type 2 diabetes mellitus is a common age-dependent disease. We discovered that male offspring of non-diabetic C57BL/6 and DBA/2 mice, called JYD mice, develop type 2 diabetes when they grow old. JYD mice show characteristics of insulin resistance, hyperglycaemia and hyperinsulinaemia in old age without obesity. We postulated that the mechanism of age-dependent type 2 diabetes in this model relates to caveolin-1 status in skeletal muscle, which appears to regulate insulin sensitivity in the mice. METHODS: We compared insulin sensitivity in aged C57BL/6 and JYD mice using glucose and insulin tolerance tests and (18)F-fluorodeoxyglucose positron emission tomography. We also determined insulin signalling molecules and caveolin proteins using western blotting, and altered caveolin-1 levels in skeletal muscle of C57BL/6 and JYD mice using viral vector systems, to examine the effect of this on insulin sensitivity. RESULTS: In 30-week-old C57BL/6 and JYD mice, the basal levels of IRS-1, Akt and peroxisome proliferator-activated receptor-gamma decreased, as did insulin-stimulated phosphorylation of Akt and insulin receptor beta. However, caveolin-1 was only increased about twofold in 30-week-old JYD mice as compared with 3-week-old mice, whereas an eightfold increase was seen in C57BL/6 mice. Downregulation of caveolin-1 production in C57BL/6 mice caused severe impairment of glucose and insulin tolerance. Upregulation of caveolin-1 in aged diabetic JYD mice significantly improved insulin sensitivity with a concomitant increase of glucose uptake in the skeletal muscle. CONCLUSIONS/INTERPRETATION: The level of skeletal muscle caveolin-1 is correlated with the progression of age-dependent type 2 diabetes in JYD mice. PMID- 18408915 TI - An assessment of the accuracy of renal blood flow estimation by Doppler ultrasound. AB - BACKGROUND: Knowledge of renal blood flow is considered important in the management of critically ill patients with acute renal failure. Renal Doppler ultrasound has been used to estimate renal blood flow. Its accuracy, however, has not been formally assessed. DESIGN: Prospective blinded animal study. SETTING: University physiology laboratory. SUBJECTS: Seven merino cross-ewes. INTERVENTIONS: We chronically implanted transit-time flow probes around the left renal artery and performed Doppler ultrasound measurements of RBF. We simultaneously recorded RBF values obtained with implanted flow probes and Doppler ultrasound during (a) observation, (b) dobutamine and (c) nitroprusside infusion in random order. RESULTS: In a total of 202 paired measurements, Doppler ultrasound measured peak systolic velocity (PSV) correlated very weakly with implanted flow probe measurements of RBF (r (2) = 0.015), as did end-diastolic velocity (EDV; r (2) = 0.086) and mean velocity (MV_vel; r (2) = 0.04). We also found similar weak correlations with other Doppler-ultrasound-derived indices. All comparisons showed bias and wide limits of agreement. CONCLUSIONS: Doppler ultrasound-derived estimates of RBF show little correlation with transit-time flow probe measurements, display significant bias and wide limits of agreement and have low accuracy for clinically significant changes in RBF in large animals. PMID- 18408916 TI - Preoperative apolipoprotein CI levels correlate positively with the proinflammatory response in patients experiencing endotoxemia following elective cardiac surgery. AB - OBJECTIVE: Experimental models show that apolipoprotein CI (apoCI) binds and enhances the inflammatory response to endotoxin. We studied in patients undergoing cardiopulmonary bypass surgery (CPB) and experiencing endotoxemia during reperfusion whether plasma apoCI levels correlate with the inflammatory response and perioperative cytokine release. DESIGN: Prospective, observational, clinical cohort study. SETTING: Operating room (OR) and intensive care unit (ICU) of a university hospital. PATIENTS: One hundred fifty-nine consecutive patients > 18 years of age (66% males =105), median age 65 and 67 years for males and females, respectively) undergoing elective cardiothoracic surgery with cardiopulmonary bypass. INTERVENTIONS: None. MEASUREMENTS: Baseline apoCI, apoCIII, total cholesterol and triglyceride levels, and perioperative endotoxin and TNF-alpha levels were determined. RESULTS: High preoperative plasma apoCI, but not apoCIII, levels were associated (p < 0.05) with increased perioperative levels of TNF-alpha in patients experiencing endotoxemia. This association was not observed in patients without endotoxemia. CONCLUSION: High plasma apoCI is positively related to proinflammatory response in patients experiencing endotoxemia and confirms the observations in animal models. PMID- 18408917 TI - Functional reactivity of the dopaminergic system following acute and chronic ketamine treatments. AB - This study examined the effects of acute (15 mg/kg, i.p.) and chronic subanesthetic (15 mg/kg, i.p., t.i.d, for 6 days) doses of ketamine [a noncompetitive N-methyl-D: -aspartate (NMDA) receptor antagonist] on amphetamine (presynaptic dopamine releasing agent; 10 mg/kg, i.p.) and apomorphine (a D(2) receptor agonist; 1 mg/kg, i.p.)-induced stereotyped behaviors. The effect of acute and chronic ketamine on haloperidol (a D(2) receptor antagonist; 1.6 mg/kg, i.p.)-induced catalepsy was also examined. Acute ketamine and chronic ketamine pretreatment increased amphetamine-induced stereotyped sniffing and locomotion compared with control groups. Acute ketamine significantly increased apomorphine induced stereotyped sniffing. However, chronic ketamine had no significant effect on apomorphine-induced stereotyped sniffing. Acute, but not chronic ketamine treatment abolished haloperidol-induced catalepsy. The increase in amphetamine induced stereotyped behaviors and the reversal of haloperidol-induced catalepsy by acute ketamine suggest that blockade of NMDA receptors by ketamine facilitates dopaminergic transmission. The absence of significant effect of chronic ketamine on apomorphine-induced stereotyped sniffing and haloperidol-induced catalepsy suggests that chronic ketamine does not modulate postsynaptic dopaminergic D(2) receptors. It is suggested that chronic ketamine increased amphetamine-induced behaviors by causing hypersensitivity of presynaptic dopamine releasing mechanisms on dopaminergic terminals. PMID- 18408918 TI - The in vivo gastrointestinal activity of TD-5108, a selective 5-HT(4) receptor agonist with high intrinsic activity. AB - The in vivo preclinical pharmacodynamic profile of TD-5108, a selective 5-HT(4) receptor agonist with high intrinsic activity, was compared to that of the clinically studied gastrointestinal pro-kinetic agents, tegaserod, cisapride and mosapride. The activity of TD-5108 was evaluated in guinea pig colonic transit, rat oesophageal relaxation and dog gastrointestinal smooth muscle contractility models. Subcutaneous administration of TD-5108, tegaserod, cisapride and mosapride increased guinea pig colonic transit (rank order of potencies: TD-5108 > tegaserod > cisapride > mosapride). Following intravenous and intraduodenal dosing, TD-5108, tegaserod, cisapride and mosapride produced dose-dependent relaxation of the rat oesophagus. On a molar basis, TD-5108 was approximately twofold less potent than tegaserod following intravenous dosing but 6- or 86-fold more potent than cisapride or mosapride, respectively, and 9- or 18-fold more potent than tegaserod or cisapride, respectively, after intraduodenal administration. Orally dosed TD-5108 increased the contractility of the canine antrum, duodenum and jejunum with higher potency than tegaserod. The selective 5 HT(4) receptor agonist, TD-5108, demonstrates robust in vivo activity in the guinea pig, rat and dog gastrointestinal tracts. PMID- 18408920 TI - Modern chemical analysis in archaeometry. PMID- 18408919 TI - Effects of selective 5-HT1A agonist tandospirone on the rate and rhythmicity of binocular rivalry. AB - RATIONALE: When dissimilar figures are presented to each eye individually, perception alternates spontaneously between each monocular view. This phenomenon, binocular rivalry, has been suggested to depend on serotonergic neural systems. OBJECTIVE: To investigate the relationship between binocular rivalry and serotonergic neural systems in more detail, we measured the perceptual alternation rate and rhythmicity of binocular rivalry under the influence of the 5-HT(1A) agonist tandospirone. MATERIALS AND METHODS: Fifteen healthy volunteers were tested under placebo and 60 mg tandospirone conditions using the single blind method. The perceptual alternation rate and rhythmicity of binocular rivalry were measured at pre-administration and at 90, 180, and 360 min post administration. RESULTS: Compared to placebo, the administration of tandospirone decreased the rate and rhythmicity of perceptual switching significantly at 90 and 180 min post-administration in a manner that is consistent with its pharmacokinetics. CONCLUSIONS: The present results suggest that the 5-HT(1A) neural pathways, mainly located in the mesolimbic system and brainstem, may play a crucial role as an oscillator in perceptual rivalry alternations. PMID- 18408921 TI - Detection of bacterial quorum sensing N-acyl homoserine lactones in clinical samples. AB - Bacteria communicate among themselves using certain chemical signaling molecules. These signaling molecules generally are N-acyl homoserine lactones (AHLs) in Gram negative bacteria and oligopeptides in Gram-positive bacteria. In addition, both Gram-positive and Gram-negative bacteria produce a family of signaling molecules known as autoinducer-2 that they employ for their communications. Bacteria coordinate their behavior by releasing and responding to the chemical signaling molecules present in proportion to their population density. This phenomenon is known as quorum sensing. The role of bacteria in the pathogenesis of several diseases, including gastrointestinal (GI) disorders, is well established. Moreover, rather recently bacterial quorum sensing has been implicated in the onset of bacterial pathogenicity. Thus, we hypothesized that the signaling molecules involved in bacterial communication may serve as potential biomarkers for the diagnosis and management of several bacteria-related diseases. For that, we previously developed a method based on genetically engineered whole-cell sensing systems for the rapid, sensitive, cost-effective and quantitative detection of AHLs in biological samples, such as saliva and stool, from both healthy and diseased individuals with GI disorders. Although various analytical methods, based on physical-chemical techniques and bacterial whole-cell biosensors, have been developed for the detection of AHLs in the supernatants of bacterial cultures, only a few of them have been applied to AHL monitoring in real samples. In this paper, we report work performed in our laboratory and review that from others that describes the detection of AHLs in biological, clinical samples, and report some of our recent experimental results. PMID- 18408922 TI - Thermal desorption characterisation of molecularly imprinted polymers. Part II: Use of direct probe GC-MS analysis to study crosslinking effects. AB - A powerful method utilising direct probe thermal desorption GC-MS is presented for the study of molecularly imprinted polymers (MIPs). A series of 2 aminopyridine (2-apy)-imprinted methacrylic acid-ethyleneglycol dimethacrylate (MAA-EGDMA) copolymers were prepared under identical conditions but with varying amounts of EGDMA (crosslinking monomer). The use of appropriate temperature programmes permitted template removal, and the subsequent assessment of polymer affinity and specificity, all of which were found to be dependent on polymer composition and morphology. The system was sufficiently sensitive to identify a specific response of imprinted polymers over nonimprinted counterparts. Correlations were found to exist between thermal desorption analysis and solution phase binding, which was assessed by UV spectroscopy, where specificity was found to diminish with decreasing EGDMA concentration. This was attributed to the increased number of free carboxyl groups in those polymers containing a lower percentage of EGDMA. Thermal desorption profiles obtained for the analyte were found to be unaffected by the physical and chemical properties of the solvent used for analyte reloading. PMID- 18408923 TI - Curative embolization of cerebral arteriovenous malformations (AVMs) with Onyx in 101 patients. AB - INTRODUCTION: Onyx has emerged in the recent years as a new embolic material. We present our experience with Onyx in the curative embolization of brain cerebral arteriovenous malformations (AVMs). METHODS: We retrospectively reviewed the files of all patients that we had treated with Onyx embolic material for a brain AVM during the last 4 years. We identified 101 patients who underwent a total of 219 sessions of embolization. RESULTS: The treatment has been concluded in 52 patients; in this group, total occlusion was obtained in 28 (53.9%) patients and near-total occlusion was obtained in 18 (34.6%). The respective figures based on the 101 patients of the study cohort are 28/101 (27.7%) and 18/101 (17.8%). More embolization sessions are to be carried out on the remaining 49 patients. There were three deaths, and eight patients had permanent neurological deficits with a resulting morbidity of 8% and mortality of 3%. CONCLUSION: High rates of total or near-total occlusion of brain AVMs can be achieved with multiple sessions of Onyx embolization with acceptable morbidity and mortality. PMID- 18408924 TI - High-resolution morphological and biochemical imaging of articular cartilage of the ankle joint at 3.0 T using a new dedicated phased array coil: in vivo reproducibility study. AB - OBJECTIVE: The objective of this study was to evaluate the feasibility and reproducibility of high-resolution magnetic resonance imaging (MRI) and quantitative T2 mapping of the talocrural cartilage within a clinically applicable scan time using a new dedicated ankle coil and high-field MRI. MATERIALS AND METHODS: Ten healthy volunteers (mean age 32.4 years) underwent MRI of the ankle. As morphological sequences, proton density fat-suppressed turbo spin echo (PD-FS-TSE), as a reference, was compared with 3D true fast imaging with steady-state precession (TrueFISP). Furthermore, biochemical quantitative T2 imaging was prepared using a multi-echo spin-echo T2 approach. Data analysis was performed three times each by three different observers on sagittal slices, planned on the isotropic 3D-TrueFISP; as a morphological parameter, cartilage thickness was assessed and for T2 relaxation times, region-of-interest (ROI) evaluation was done. Reproducibility was determined as a coefficient of variation (CV) for each volunteer; averaged as root mean square (RMSA) given as a percentage; statistical evaluation was done using analysis of variance. RESULTS: Cartilage thickness of the talocrural joint showed significantly higher values for the 3D-TrueFISP (ranging from 1.07 to 1.14 mm) compared with the PD-FS-TSE (ranging from 0.74 to 0.99 mm); however, both morphological sequences showed comparable good results with RMSA of 7.1 to 8.5%. Regarding quantitative T2 mapping, measurements showed T2 relaxation times of about 54 ms with an excellent reproducibility (RMSA) ranging from 3.2 to 4.7%. CONCLUSION: In our study the assessment of cartilage thickness and T2 relaxation times could be performed with high reproducibility in a clinically realizable scan time, demonstrating new possibilities for further investigations into patient groups. PMID- 18408925 TI - Kinetics of anti-carcinoembryonic antigen antibody internalization: effects of affinity, bivalency, and stability. AB - Theoretical analyses suggest that the cellular internalization and catabolism of bound antibodies contribute significantly to poor penetration into tumors. Here we quantitatively assess the internalization of antibodies and antibody fragments against the commonly targeted antigen carcinoembryonic antigen (CEA). Although CEA is often referred to as a non-internalizing or shed antigen, anti-CEA antibodies and antibody fragments are shown to be slowly endocytosed by LS174T cells with a half-time of 10-16 h, a time scale consistent with the metabolic turnover rate of CEA in the absence of antibody. Anti-CEA single chain variable fragments (scFvs) with significant differences in affinity, stability against protease digestion, and valency exhibit similar uptake rates of bound antibody. In contrast, one anti-CEA IgG exhibits unique binding and trafficking properties with twice as many molecules bound per cell at saturation and significantly faster cellular internalization after binding. The internalization rates measured herein can be used in simple computational models to predict the microdistribution of these antibodies in tumor spheroids. PMID- 18408926 TI - Molecular mechanisms of MHC class I abnormalities and APM components in human tumors. AB - Tumor immune escape plays a critical role in cancer, but the mechanisms involved in this process have still to be defined. In the recent years, progress has been made in understanding how peptides presented by MHC class I molecules were generated, in particular which proteases are involved in this process and how intracellular pathways influence antigen presentation in professional antigen presenting cells and in various types of malignancies. Different MHC class I abnormalities have been found in solid tumors of distinct origin, but also in hematopoietic diseases. These include structural alterations such as total, haplotype and allelic loss of the MHC class I heavy chain, deletions and point mutations, in particular in beta2-microglobulin and TAP1 as well as dysregulation of various components of the MHC class I antigen processing machinery (APM), which could occur at the epigenetic, transcriptional and posttranscriptional level. The lack or downmodulation of the expression of single or multiple components of the MHC class I antigen processing pathway may avoid the recognition of tumor cells by tumor-specific CD8+ cytotoxic T lymphocytes. This review will give an overview of the underlying molecular mechanisms of MHC class I abnormalities in human tumors of distinct histology, which also might have an impact on the design of T cell-based immunotherapies. PMID- 18408927 TI - Anterior corpectomy and fusion for severe ossification of posterior longitudinal ligament in the cervical spine. AB - Between May 2002 and October 2006, 19 patients (17 men and 2 women; average age 57.2; range 47-71 years) received anterior corpectomy and fusion for severe ossification of the posterior longitudinal ligament (OPLL) in our department. Preoperative radiological evaluation showed the narrowing by the OPLL exceeded 50% in all cases, and OPLL extended from one to three vertebrae. We followed-up all patients for 12-36 months (mean 18 months). The Japanese Orthopaedic Association (JOA) score before surgery was 9.3 +/- 1.8 (range 5-12) which significantly increased to 14.2 +/- 1.3 (range 11-16) points at the last follow up (P < 0.01). The improvement rate (IR) of neurological function ranged from 22.2-87.5%, with a mean of 63.2% +/- 15.2%. The operation also provided a significant increase in the cervical lordosis and the cord flatting rate (P < 0.01). No severe neurological complication developed. We therefore concluded that anterior decompression and fusion was effective and safe in the treatment of the selected patients, although OPLL exceeded 50% diameter of the spinal canal. PMID- 18408928 TI - Extramedullary plasmacytomas of the thyroid and pericardium as initial presentation of multiple myeloma. PMID- 18408929 TI - Autoimmune retinopathy: a review and summary. AB - Three main forms of autoimmune retinopathy (AIR) have been identified over the last 15 years: cancer-associated retinopathy (CAR), melanoma-associated retinopathy (MAR), and nonneoplastic autoimmune retinopathy (npAIR). In this chapter, the term AIR will be used to encompass all three disorders where there is commonality to their features. Complicating the issue is that AIR can be a secondary complication of other conditions such as retinitis pigmentosa, ocular trauma, birdshot retinopathy, acute zonal occult outer retinopathy (AZOOR), or multiple evanescent white dot syndrome (MEWDS). The many forms of AIR tend to have common clinical features despite the fact that there has been no uniform set of anti-retinal antibodies circulating in these patients. Patients tend to have a wide variance of anti-retinal antibody activity often with three to six different antibodies found on immunoblots. Patients typically present with a sudden onset of photopsia, rapid visual loss, and abnormal electroretinograms (ERGs). Most patients have a panretinal degeneration without pigment deposits. PMID- 18408930 TI - A codon-optimized luciferase from Gaussia princeps facilitates the in vivo monitoring of gene expression in the model alga Chlamydomonas reinhardtii. AB - The unicellular green alga Chlamydomonas reinhardtii has emerged as a superb model species in plant biology. Although the alga is easily transformable, the low efficiency of transgene expression from the Chlamydomonas nuclear genome has severely hampered functional genomics research. For example, poor transgene expression is held responsible for the lack of sensitive reporter genes to monitor gene expression in vivo, analyze subcellular protein localization or study protein-protein interactions. Here, we have tested the luciferase from the marine copepod Gaussia princeps (G-Luc) for its suitability as a sensitive bioluminescent reporter of gene expression in Chlamydomonas. We show that a Gaussia luciferase gene variant, engineered to match the codon usage in the Chlamydomonas nuclear genome, serves as a highly sensitive reporter of gene expression from both constitutive and inducible algal promoters. Its bioluminescence signal intensity greatly surpasses previously developed reporters for Chlamydomonas nuclear gene expression and reaches values high enough for utilizing the reporter as a tool to monitor responses to environmental stresses in vivo and to conduct high-throughput screenings for signaling mutants in Chlamydomonas. PMID- 18408931 TI - Molecular analysis of hormone-regulated petal regeneration in Petunia. AB - The petal is an important floral organ of higher plants. To study the mechanism of petal development, the in vitro regeneration system of petals was established in Petunia. High-frequency induction of petals occurred directly from explants on the media containing the combination of N6-benzyladenine (6-BA) and indole-3 acetic acid (IAA). Expression analysis of genes involved in flower development indicated that these genes were classified into three types. ABERRANT LEAF AND FLOWER (ALF) gene was induced during petal regeneration. Whereas, B-class and E class genes, and genes involved in cell division were constitutively upregulated. In contrast, C-class and D-class genes were not expressed in explants and regenerated tissues. Further, in situ hybridization analysis showed that both ALF and GREEN PETAL (GP) expression were spatially regulated. The results suggest that differential regulation of gene expression occurs in the presence of hormones during petal regeneration, and hormone-regulated gene expression might be required for petal regeneration. This study provides the preliminary information to understand the mechanism of petal regeneration. PMID- 18408932 TI - What is the molecular pathology of low-risk prostate cancer? AB - OBJECTIVES: Low-risk prostate cancer is defined as a clinical T1c or T2a tumor with a Gleason score of 0.05). One patient in the first group had intestinal obstruction and intestinal perforation. For the second group, early complications were related to the colostomy. At after 3 years postoperatively, 9 patients in the first group and 12 patients in the second group were followed-up and all had positive voluntary bowel movements. There were no statistically significant differences in soiling and constipation grades between the two groups. The total length of hospital stay was 12.06 +/- 0.85 and 33.85 +/- 0.94 days and the cost was 10,681.1 +/- 1,759.5 and 27,355.9 +/- 1,952.0 RMB for the first and second groups, respectively. There was a statistically significant difference in the total length of hospital stay and cost between the two groups (P < 0.0001); however, there were no statistically significant differences in the length of hospital stay and cost during the definitive operation between the two groups (P > 0.05). This retrospective study shows that it is feasible for correction of imperforate anus with a rectourethral or a rectovestibula fistula in neonates using a modified PSARP without a colostomy. PMID- 18408937 TI - Management and surgical treatment for focal nodular hyperplasia in children. AB - Focal nodular hyperplasia (FNH) is rare in pediatric liver tumor. To investigate the experience in the appropriate management of pediatric FNH, the records of consecutive children who were managed at our institute from 2000 to 2007were reviewed for FNH. There were four males and nine females whose ages ranged from 26 months to 18 years with a mean of 11.7 years. FNH in most children was detected by medical examination for abdominal pain (61.5%) or other symptoms. All the 13 pediatric patients underwent liver resection. There was no operative death or postoperative complications. The children were regularly followed up ranging from 2 to 85 months and they were healthy without recurrence. For pediatric FNH patients with clinical symptoms or indefinite diagnosis, it is suggested that active surgical treatment by hepatectomy should be performed. PMID- 18408938 TI - A comparison of analgesic efficacy of tramadol and pethidine for management of postoperative pain in children: a randomized, controlled study. AB - Prevention of postoperative pain in children is one of the most important objectives of the anesthesiologist. Opioids have been used as an analgesic for postoperative pain in children for many years. Tramadol has both opioid and monoaminergic agonist actions. The aim of the study was to determine if the analgesic potency and occurrence of adverse effects of tramadol differ from pethidine when administered to children. A total of 110 healthy children, aged 2 12 years, scheduled for elective lower abdominal surgery were randomized to receive either pethidine 1 mg/kg (Group I, n = 60) or tramadol 2 mg/kg (Group II, n = 50) for postoperative pain after anesthesia induction. Pain intensity, adverse effects, heart rate, and systolic and diastolic blood pressure were recorded at regular intervals. The mean pain scores on postoperative 24 h were significantly greater with tramadol than with pethidine. Sedation scores, heart rate and systolic and diastolic blood pressure showed no significant differences between the groups. We conclude that pethidine and tramadol are effective in providing analgesia in pediatric patients, but pethidine provided better postoperative analgesia than tramadol. Changes in blood pressure, heart rate and arterial oxygen saturation were minimal and were similar in both drugs. PMID- 18408939 TI - Uro-genital bleeding in pre-menarcheal girls: dilemmas of child abuse. AB - Uro-genital bleeding in pre-menarcheal girls always raises concerns regarding child abuse. There are serious social issues involved; however, before labeling it as sexual abuse, all efforts must be made to rule out an organic pathology. We recently encountered two young girls of Afro-Caribbean origin who presented with vaginal bleeding with concerns from referring physicians about child/sexual abuse. On examination both had a rare condition of urethral prolapse. One of them underwent four-quadrant excision and the other was treated conservatively with urethral catherization. PMID- 18408940 TI - A simplified technique for giant inguinal hernia repair in infants. AB - Repair of giant inguino-scrotal hernia (GISH) in male infants is a difficult operation, even in experienced hands. It requires an immaculate technique to avoid known complications such as tearing of the sac, injury to delicate testicular vessels and dividing of vas deferens. Moreover, a recurrence rate of 9% is noted in a number of reports. This article describes a new surgical maneuver to simplify the procedure. All GISH repaired by the author, over a 5 year period (October 2001-September 2006), were reviewed retrospectively. In total, 89 infants with 106 GISH underwent uni- or bilateral herniotomies. A standard inguinal incision is made and Scarpa's fascia is sharply opened; the external inguinal ring and the cord is identified. By gentle manipulation and blunt dissection, the spermatic cord together with the testis is exteriorized. The assistant applies gentle traction to the cord, which allows for easy identification of the inguinal sac and its subsequent separation from vas and vessels. Testis is replaced in the scrotum, hernial sac suture ligated at its base and the wound closed in layers. All cases were managed with the above approach. The average length of the procedure was 11 min for unilateral and 19 min for bilateral cases. Except for minimal scrotal swelling post-operatively, no other surgery-related complications were noted during or immediately after the operation. Testicular atrophy or iatrogenic undescended testes were not encountered in the follow-up period. Ipsilateral recurrent hernia was noted in one infant after 6 months which required re-operation with the same technique. In cases of GISH; dislocating the testis into the wound and applying a gentle stretch on the cord allows for a safe dissection of the hernial sac and subsequent herniotomy. This maneuver converts a difficult procedure into a relatively simple one. PMID- 18408941 TI - Study of acetylcholinesterase activity in rectal suction biopsy for diagnosis of intestinal dysganglionoses: 17-year experience of a single center. AB - Although the utility of the acetylcholinesterase (AChE) histochemistry on rectal suction biopsy in diagnosing Hirschsprung's disease (HD) has been documented, few reports address a great number of biopsies and patients. Our aim is to present a 17-year experience on the method of rectal suction biopsy and AChE histochemical staining for diagnosis of intestinal dysganglionoses. Between August 1989 and July 2006, 297 children suspected of having HD were submitted to rectal suction biopsies that were evaluated by the same two surgeons. There were 18 complications (6.0%), namely one self-limited rectal bleeding and 17 (5.7%) inadequate procedures that were repeated. A total of 157 patients (52.8%) showed no increased AChE activity and the remaining patients (140-47.2.0%) presented patterns of increased AChE activity confirming the diagnosis of HD or neuronal intestinal dysplasia. Among the 140 cases suspected as having HD, in 131 children the diagnosis of HD was confirmed and they were operated on. The histological studies showed that 111 children presented the classic form of HD or a long spastic segment. Sixteen children presented total colonic aganglionosis and four children proved to have intestinal neuronal dysplasia, according to histological and radiological criteria. Nine (6.6%) newborns were identified as false positives and no false-negative results were verified. The rectal suction biopsy combined with AChE staining is advantageous for the differentiation between normal bowel and intestinal dysganglionoses. The rectal suction method is simple and can easily be performed by experienced surgeons. The histological evaluation is very objective and can be performed by a non-pathologist. PMID- 18408942 TI - Antegrade continence enema (ACE): current practice. AB - The purpose of this study was to assess current status of antegrade continence enema (ACE) procedure taking into account the recent improvement in the technique and outcome. Reviewing our record of 48 patients with ACE procedure performed between January 2002 and May 2007, we found that the underlying diagnoses were idiopathic constipation in 56%, anorectal malformation in 31%, spina bifida in 8% and Hirschsprung's disease in 4%. Mean age of operation was 10.7 years. Appendix was used as stoma in 73% of cases. Stomal stenosis requiring revision was seen in 6% of cases and continence was achieved in 92% of cases. A systematic search of database was performed for the same period. Twenty-four studies describing 676 patients were found. The mean age was 10 years and various sites used for ACE were, right side of abdomen in 71%, umbilicus in 15% and left side of abdomen in 14%. The incidence of open and laparoscopic procedures were 87 and 13%, respectively. Appendix was used for stoma in 76% procedures. Other operative modalities were retubularised colon, retubularised ileum, caecal button and caecostomy tube, etc. The mean volume of enema fluid used was 516 ml. The mean evacuation time was 42 min. Stomal stenosis requiring revision was seen in 13% of cases. Continence was achieved in 93% of cases. There has been significant improvement in the outcome during last 5 years in comparison to the outcome published in late 1990s. Advancements in techniques, better-trained stoma care nurses and better stoma appliances could have played major role in this success. PMID- 18408943 TI - [Blue sclerae syndrome]. AB - The medical history of a 46-year-old female patient with dark-blue sclerae showed repeated fractures of the extremity skeleton and clavicular following minor trauma up to the age of 15. The sclerae indicated osteogenesis imperfecta (OI), which leads to disrupted collagen synthesis due to various mutations of the type 1 procollagen gene, which in turn leads to brittle bones with reduced bone density and greater susceptibility to fracture. In OI, distinction is made between varying clinical types which show differing degrees of severity. Bisphosphonates, which can reduce significantly increased bone turnover, are a very promising therapy approach. PMID- 18408944 TI - Acute tibial tubercle avulsion fractures in the sporting adolescent. AB - INTRODUCTION: The authors report 7 cases of acute tibial tubercle avulsion fractures. The fracture occurred in 6 out of the 7, after an abrupt tension of the patellar tendon in male sporting adolescents (age 13-17 years). Two patients presented symptoms of homolateral Osgood-Schlatter's disease before the lesion. METHOD: According to Ogden's classification, the tibial tubercle avulsion fracture was not displaced in 3 cases (stage IA) and was treated conservatively by immobilization for 6 weeks. In 4 cases, the fracture was displaced and necessitated an internal fixation with plaster for about 6 weeks. A torn patellar tendon was noted in one adolescent having a stage IIIB avulsion fracture. RESULT: The mean follow-up was of 4.5 years (1.5-7.5 years). The results were satisfactory: complete functional recovery, resumption of sport at the previous level and absence of recurvatum. PMID- 18408945 TI - Transpedicular plate fixator as effective system of spine stabilisation: biomechanical characteristics. AB - INTRODUCTION: Zespol fixator, which was created in Poland by Ramatowski and Granowski, has an angular stable connection of screws and plate. These properties of this plate fixator, that is effective and not an expensive system of osteosynthesis of shaft of long bone widely used in Poland, impelled us to adapt it as a transpedicular plate fixator of spine. AIM: The aim of our in vitro study was to measure loads acting on spine stabilized by transpedicular plate fixator and to determine if its stability is comparable with uninjured spine. We also hypothesized that the spine stability with examined fixator had similar properties as spine fixators constructed with screws and rods. MATERIALS AND METHODS: We tested its biomechanical properties and compared it with a CD device by using specimens of four human spines. Each spine with damage induced in laboratory conditions was stabilised by one of those stabilisers in one (L4-L5) or two (Th12-L2) motion segments and subsequently were subject to load. The spines without and with one of transpedicular stabilization were subject to an unsymmetrical shift of +3/-4 mm for extension-compression and symmetrical shift for bending, in the frontal plane (+0.14/-0.14 rad) and the sagittal plane (+0.11/-0.11 rad), respectively. RESULTS: Loads during extension-compression and bending in the sagittal plane were similar to the uninjured spine for short stabilization by using both stabilizers and amounted to 92.3 and 98.26%, respectively, of the load range sums of healthy spines. For long stabilization these loads amounted to 93.2 and 84.4%, respectively. Only following short and long stabilization for both devices in case of bending in the frontal plane the increase in loads up to 144.2 and 163.3% of the range sums of uninjured spines was achieved. CONCLUSION: It corroborates the fact that the application of the modified Zespol device for spine stabilisation provides the possibility of restoring its load transfer capacity similar to that in the healthy spine and comparable with the CD device. PMID- 18408946 TI - Comparison between the Percutwist and the Ciaglia percutaneous tracheotomy techniques. AB - A prospective study was designed to compare two single-dilator percutaneous tracheotomy techniques, the Ciaglia BlueRhino and the Percutwist technique. One hundred and ninety adult patients were included, 166 with the BlueRhino, a conical shaped, flexible rubber dilator, and 24 with the Percutwist, a screw like dilating device. The procedure was performed under fiberscopy in the intensive care unit (ICU). Age, body mass index (BMI), indication for tracheotomy, surgical landmarks, duration of the procedure and surgical complications were recorded. Median age and indications were similar for the two groups. Dilation was successful in all patients. The mean time for surgery was shorter with the Ciaglia technique: 8 +/- 3 versus 12 +/- 5 min with the Percutwist technique (P = 0.004). There was no significant difference related to weight, BMI, duration of tracheotomy and complications between both groups. One posterior tracheal wall puncture was observed with the Ciaglia technique and four with the Percutwist technique. No serious complications were noted with either technique. The Percutwist technique represents an alternative to the more established Ciaglia BlueRhino technique. The Ciaglia technique is a safe and more rapid procedure for bedside tracheotomy. PMID- 18408947 TI - Transcription-dependent spatial arrangements of CFTR and conserved adjacent loci are not conserved in human and murine nuclei. AB - The human genes CFTR, ASZ1/GASZ, and CTTNBP2/CORTBP2 map to adjacent loci on chromosome 7q31 and display characteristic patterns of nuclear positioning, which strictly correlate with the state of activity. To address the evolutionary conservation of gene positioning, we investigated transcriptional activity and nuclear positioning of the highly conserved murine orthologs and of additional murine genes mapping to the region of conserved synteny on mouse chromosome 6. The results showed that all murine loci investigated constitutively localized in the nuclear interior irrespective of their functional state. Silenced loci did not display preferential association with the nuclear periphery or with chromocenters, respectively, and no differential positioning with respect to the chromosome 6 territory could be observed. This positional behavior of the murine loci was in striking contrast to the positioning of the human orthologs, and the results show that the transcription-dependent positioning of CFTR and adjacent loci has not been conserved. The findings reveal that the nuclear organization of conserved chromosomal regions can change rapidly during evolution and is not always as highly conserved as other features of chromosome organization. Furthermore, the results suggest that the way how nuclear positioning contributes to the regulation of conserved loci can be different in different vertebrate species. PMID- 18408948 TI - Comparisons between five self-administered instruments predicting sick leaves in a 4-year follow-up. AB - OBJECTIVES: This study aimed to explore and compare the ability of five instruments for self-rating to predict future sick leave rates. METHODS: In three Swedish municipalities 2,252 employees completed a baseline questionnaire and were followed up for 4 years. Five health-oriented instruments for self-rating were used as potential predictors of the two outcome measures no sick leave at all, and one or more spells of long-term sick leave >or=28 days. Positive and negative predictive values as well as Cox proportional hazard ratios (denoted as RRs) adjusted for age and work type were calculated. RESULTS: The instruments showed no statistical difference in predicting future sick leave for either of the sexes. For no sick leave RRs ranged between 1.27 and 1.52 (women), 1.35 and 1.61 (men); for long-term sick leave RRs ranged between 1.78 and 2.39 (women), 2.87 and 5.53 (men). However, the best prediction of long-term sick leave for men, RR 5.53, 95% confidence interval (CI) 3.37-9.08, was significantly higher than the best prediction for women, RR 2.39, 95% CI 1.97-2.90. CONCLUSION: Prediction of long-term sick leave was better than that of no sick leave, and better among men than among women. There was a tendency for somewhat better prediction of future sick leave by multiple-question instruments, but single question instruments can very well be used in predicting future sick leaves, and crude analyses stratified by sex can be used for screening purposes. PMID- 18408949 TI - Lung function predicts survival in a cohort of asbestos cement workers. AB - PURPOSE: To study the predictive power of respiratory screening examinations a cohort of asbestos workers was followed from active work in an asbestos cement plant until death. METHODS: From a cohort with data on individual exposure since first employment 309 workers who had a preventive medical examination in 1989/1990 were observed until death or the end of 2006. The impact of asbestos exposure (fibre years) and of smoking history on lung function was examined by linear regression, on specific causes of death and total mortality by Cox regression. The prognostic value of lung function, chest X-ray, and various clinical findings regarding total mortality was also examined by Cox regression. RESULTS: Lung function proved to be the best predictor of survival apart from current smoking. Depending on the lung function variable an impairment by the interquartile range resulted in a hazard ratio of 1.5-1.6 while for current smokers it was 2.3. An increase of 70 fibre years (interquartile range) led to a hazard ratio of only 1.1. Lung function was influenced by asbestos exposure, current (but not former) smoking, and by pathological X-ray findings. The risk for pleural mesothelioma was dominated by time since first exposure to crocydolite in the pipe factory while the risk for bronchial cancer increased with smoking and total fibre years. An unexpected finding was an increase of gastric cancer in asbestos cement workers. CONCLUSION: Lung function decrease predicts risk of premature death better than exposure history and regular spirometry should therefore be offered as primary screening to all former asbestos workers. In workers with a history of high cumulative exposure or rapid lung function decrease or radiological signs (diffuse pleural thickening or small irregular opacities) more sensitive techniques (high resolution computer tomography) need to be applied. All smokers with a history of asbestos exposure should be given free smoking cessation therapy to prevent premature death and lung cancer in particular. PMID- 18408950 TI - The trainability and contralateral response of cold-induced vasodilatation in the fingers following repeated cold exposure. AB - Cold-induced vasodilatation (CIVD) is proposed to be a protective response to prevent cold injuries in the extremities during cold exposure, but the laboratory based trainability of CIVD responses in the hand remains equivocal. Therefore, we investigated the thermal response across the fingers with repeated local cold exposure of the whole hand, along with the transferability of acclimation to the fingers of the contralateral hand. Nine healthy subjects immersed their right hand up to the styloid process in 8 degrees C water for 30 min daily for 13 days. The left hand was immersed on days 1 and 13. Skin temperature was recorded on the pads of the five fingertips and the dorsal surface of the hand. The presence of CIVD, defined as an increase in finger skin temperature of 0.5 degrees C at any time during cooling, occurred in 98.5% of the 585 (9 subjects x 5 sites x 13 trials) measurements. Seven distinct patterns of thermal responses were evident, including plateaus in finger temperature and superimposed waves. The number (N) of CIVD waves decreased in all digits of the right hand over the acclimation period (P = 0.02), from average (SD) values ranging from 2.7 (1.7) to 3 (1.4) in different digits on day 1, to 1.9 (0.9) and 2.2 (0.7) on day 13. Average (SD) finger skin temperature (T (avg)) ranged from 11.8 (1.4) degrees C in finger 5 to 12.7 (2.8) degrees C in finger 3 on day 1, and then decreased significantly (P < 0.001) over the course of the training immersions, attaining values ranging from 10.8 (0.9) degrees C in finger 4 to 10.9 (0.9) degrees C in finger 2 on day 13. In the contralateral hand, N was reduced from 2.5 to 1.5 (P < 0.01) and T (avg) by approximately 2 degrees C (P < 0.01). No changes were observed in thermal sensation or comfort of the hand over the acclimation. We conclude that, under conditions of whole-hand immersion in cold water, CIVD is not trainable and may lead to systemic attenuation of thermal responses to local cooling. PMID- 18408951 TI - Enhancement of cold-induced vasodilatation following acclimatization to altitude. AB - The present study evaluated the effect of high-altitude acclimatisation on the cold-induced vasodilatation (CIVD) response. A group of highly trained mountaineers (N = 9; Alpinists) were tested before and after a 3 week high altitude Himalayan expedition (altitude ranging from 3,985 to 6,828 m). A control group (N = 7) with no mountaineering experience was tested at the same time points. During each test, subjects first immersed their hand to the styloid process in 37 degrees C water for 5 min and then in 10 degrees C water for 30 min. Upon completion of the hand immersion, the same procedure was repeated for the foot. Skin temperature of the pads of all immersed digits was measured throughout the immersion and for 10 min following the immersion. In the Alpinists, a significant increase in amplitude of CIVD and absolute maximum finger skin temperature during immersion was observed in the hand post expedition. For the foot, peak time of CIVD was significantly shorter in the Alpinist group, and there were significant increases in minimum and maximum toe skin temperature during CIVD, mean toe skin temperature during immersion, absolute minimum and maximum toe skin temperature during immersion, and absolute amplitude during immersion. The results demonstrate a significant enhancement of the CIVD response as a consequence of a brief high altitude acclimatisation, and that these changes were especially prominent in the toes. PMID- 18408952 TI - Enema-induced severe hyperphosphatemia in children. AB - We report the case of a 13-year-old boy with Costello syndrome and chronic constipation who received phosphate-containing Fleet-pediatric enemas at regular intervals. The day before admission he was given four enemas for severe constipation. Within 24 h the boy had developed severe hyperphosphatemia (phosphate 17.75 mmol/L), hypertonic dehydration (sodium 171 mmol/L) and severe hypocalcemia (calcium 0.56 mmol/L). With early intervention and treatment, the child survived without sequelae. Osmotically acting hypertonic phosphate enemas may result in severe toxicity. Children with developmental delay often display bowel dysfunction and chronic constipation and are therefore high-risk patients. The accepted opinion-that these enemas are not absorbed and therefore systemically inactive-is not true. Physicians should be aware of the potentially lethal complications of this treatment, which is part of everyday clinical practice. PMID- 18408953 TI - Acute fatty liver of pregnancy and neonatal long-chain 3-hydroxyacyl-coenzyme A dehydrogenase (LCHAD) deficiency. AB - Here we report a 7-month-old girl with long-chain 3-hydroxyacyl-coenzyme A dehydrogenase (LCHAD) deficiency with hypoketotic hypoglycemia; the mother had a history of acute fatty liver in a previous pregnancy leading to fetal death at 34 weeks of gestation. The misense mutation 1528G > C was detected in both alleles in the proband and in one allele in both parents. We emphasize that screening for fatty acid oxidation disorders and specifically LCHAD deficiency should be performed in newborns from mothers with hepatic complications during pregnancy such as acute fatty liver of pregnancy or severe or recurrent HELLP syndrome. PMID- 18408955 TI - Vacuolar-type H(+)-ATPase with the a3 isoform is the proton pump on premature melanosomes. AB - The melanosome, an organelle specialized for melanin synthesis, is one of the lysosome-related organelles. Its lumen is reported to be acidified by vacuolar type H(+)-ATPase (V-ATPase). Mammalian V-ATPase exhibits structural diversity in its subunit isoforms; with regard to membrane intrinsic subunit a, four isoforms (a1-a4) have been found to be localized to distinct subcellular compartments. In this study, we have shown that the a3 isoform is co-localized with a melanosome marker protein, Pmel17, in mouse melanocytes. Acidotropic probes (LysoSensor and DAMP) accumulate in non-pigmented Pmel17-positive melanosomes, and DAMP accumulation is sensitive to bafilomycin A1, a specific inhibitor of V-ATPase. However, none of the subunit a isoforms is associated with highly pigmented mature melanosomes, in which the acidotropic probes are also not accumulated. oc/oc mice, which have a null mutation at the a3 locus, show no obvious defects in melanogenesis. In the mutant melanocytes, the expression of the a2 isoform is modestly elevated, and a considerable fraction of this isoform is localized to premature melanosomes. These observations suggest that the V-ATPase keeps the lumen of premature melanosomes acidic, whereas melanosomal acidification is less significant in mature melanosomes. PMID- 18408954 TI - TNF receptor-associated periodic syndrome (TRAPS): description of a novel TNFRSF1A mutation and response to etanercept. AB - TRAPS is the most common of the autosomal dominant periodic fever syndromes. It is caused by mutations in the TNFRSF1A gene, which encodes for the type 1 TNF receptor (TNFR1). We describe here a Brazilian patient with TRAPS associated to a novel TNFRSF1A de novo mutation and the response to anti-TNF therapy. The patient is a 9-year-old girl with recurrent fevers since the age of 3 years, usually lasting 3 to 7 days, and recurring every other week. These episodes are associated with mild abdominal pain, nausea, vomiting and generalized myalgia. Recurrent conjunctivitis and erysipela-like skin lesions in the lower limbs also occur. Laboratory studies show persistent normocytic normochromic anemia, thrombocytosis, elevated erythrocyte sedimentation rate and C-reactive protein. IgD levels are normal. Mutational screening of TNFRSF1A revealed the association of a novel C30F mutation with the common R92Q low-penetrance mutation. The R92Q mutation is seen in 5% of the general population and is associated with an atypical inflammatory phenotype. The patient had a very good response to etanercept, with cessation of fever and normalization of inflammatory markers. Our report expands the spectrum of TNFRSF1A mutations associated with TRAPS, adding further evidence for possible additive effects of a low-penetration R92Q and cysteine residue mutations, and confirms etanercept as an efficacious treatment alternative. PMID- 18408956 TI - Detection of phenolic compounds using impedance spectroscopy measurements. AB - Langmuir-Blodgett (LB) and layer-by-layer films (LbL) of a PPV (p phenylenevinylene) derivative, an azo compound and tetrasulfonated phthalocyanines were successfully employed as transducers in an "electronic tongue" system for detecting trace levels of phenolic compounds in water. The choice of the materials was based on their distinct electrical natures, which enabled the array to establish a fingerprint of very similar liquids. Impedance spectroscopy measurements were taken in the frequency range from 10 Hz to 1 MHz, with the data analysed with principal component analysis (PCA). The sensing units were obtained from five-layer LB films of (poly[(2-methoxy-5-n-hexyloxy)-p phenylenevinylene]), OC(1)OC(18)-PPV (poly(2-methoxy,5-(n-octadecyl)-p phenylenevinylene)), DR (HEMA-co-DR13MA (poly-(hydroxyethylmethacrylate-co-[4' [[2-(methacryloyloxy)-ethyl]ethylamino]-2-chloro-4-nitroazobenzene]))) and five bilayer LbL films of tetrasulfonated metallic phthalocyanines deposited onto gold interdigitated electrodes. The sensors were immersed into phenol, 2-chloro-4 methoxyphenol, 2-chlorophenol and 3-chlorophenol (isomers) solutions at 1 x 10( 9) mol L(-1), with control experiments carried out in ultra pure water. Samples could be distinguished if the principal component analysis (PCA) plots were made with capacitance values taken at 10(3) Hz, which is promising for detection of trace amounts of phenolic pollutants in natural water. PMID- 18408957 TI - Resistance, serotype and genetic diversity of Streptococcus pneumoniae-resistant strains isolated in the West Pomerania region of Poland in the years 2001-2005. AB - The aim of this study was to analyse the resistance patterns, serotypes and genetic diversity of Streptococcus pneumoniae-resistant strains isolated in the West Pomerania region of Poland. They were clinical isolates obtained during a 5 year study (2001-2005) mainly from ambulatory patients with upper respiratory tract infections. The strains showed resistance to 8 out of 9 tested antibiotics (except vancomycin) and 53.8% of the strains were multidrug-resistant (MDR). The increase over time in the number of MDR strains and in resistance degrees was not statistically significant. Resistance to cotrimoxazole was the most frequent (86.7%). Penicillin nonsusceptibility was shown in 38% of the strains and resistance to macrolides in 36.7% of the strains, mainly of MLS(B) phenotype (94.1%). A significant resistance increase was only observed for beta-lactam antibiotic. The population of S. pneumoniae-resistant strains in our region presented 31 resistance patterns, 13 serotypes and a high genetic diversity-70 pulse field gel electrophoresis (PFGE) profiles have been described: 44 of them were unique and 26 clusters consisted of 2 to 30 strains similar by more than 87%. Cluster I, grouping 30 strains of similar resistant patterns (TSH: 70%, SH, TH, T, H, S) and mainly serotype 19F, isolated over the 5 years of the study, could represent a new national clone. The polysaccharide 23-valent vaccine covers 83.5%, while the conjugated 7-, 9- and 11-valent vaccines cover 79.1-79.7% of the resistant strains collected in our region. A statistically significant decrease of vaccine coverage in time has been noted. PMID- 18408958 TI - Fat utilization in healthy subjects consuming diacylglycerol oil diet: dietary and whole body fat oxidation. AB - Several studies in animals and humans have reported beneficial effects of diacylglycerol (DAG) on lipid and energy metabolism. We assessed the effect of DAG versus triacylglycerol (TAG) treatment on total energy expenditure (TEE), total fat oxidation (Fox) and respiratory quotient (RQ), and measured the oxidation rate of each oil using a respiratory chamber and the 13C-stable isotope. Eleven healthy subjects participated in a double-blind, randomized crossover study. Subjects consumed an energy maintenance diet consisting of 55% of total calories from carbohydrate, 15% from protein and 30% from fat during both the 3-day pre-chamber and 36-h chamber period. Fifty percent of the fat was test oil, containing either DAG oil or TAG oil. The oxidation rate of ingested test oils was determined by monitoring 13CO2 excretion in the breath from 13C labeled diolein or 13C-labeled triolein. There were no significant differences in TEE, RQ and total Fox between the DAG and TAG treatment in the overall analysis. In the subgroup analysis, DAG treatment decreased RQ significantly in subjects with a high fat ratio (HFR) compared to TAG treatment. In addition, ingested diolein oxidation in DAG treatment was significantly faster than triolein oxidation in TAG treatment in the HFR group. Enhanced fat utilization with DAG treatment and rapid oxidation of ingested DAG may, at least in part, explain the greater loss of body weight and body fat related to DAG consumption found in the weight-loss studies. PMID- 18408959 TI - The digestibility and accumulation of dietary phytosterols in Atlantic salmon (Salmo salar L.) smolt fed diets with replacement plant oils. AB - Phytosterols occur in high concentration in canola (Brassica napus L.) and other vegetable oils such as from the borage plant Echium (Echium plantagineum L.). We investigated if Atlantic salmon (Salmo salar) digest and accumulate dietary phytosterols in significant amounts in muscle and liver. Phytosterols are lipid soluble, lower cholesterol and reduce the risk of coronary heart disease in humans. We aimed to determine if fatty fish, such as salmon, can be used as a delivery source of this functional food component. Three diets containing canola oil (CO), Echium oil (EO) and fish oil (FO) were fed to Atlantic salmon smolt over 9 weeks. The digestibility of natural abundances of phytosterols by Atlantic salmon was poor compared to cholesterol. However, phytosterols accumulated in liver and muscle of fish. Significantly increased concentrations of 24 methylenecholesterol, campesterol, beta-sitosterol and total phytosterol occurred in livers of EO fed fish compared to FO fed fish. Campesterol concentrations increased in CO fed fish compared to the FO fed fish. We demonstrated that natural abundances of dietary phytosterols are digested by and accumulated in liver and white muscle of Atlantic salmon smolt. However, phytosterol levels in salmon muscle will not be a major source of phytosterols in human diets and would not be expected to significantly effect human cardiovascular health. PMID- 18408960 TI - Breast cancer in Chinese women younger than age 40: are they different from their older counterparts? AB - BACKGROUND: Breast cancer in young women is uncommon, but when it does occur it has been reported to have aggressive biological characteristics. The incidence of breast cancer peaks at age 40 in Hong Kong Chinese women, earlier than in Caucasians. This study is the first to report the tumor characteristics and management of breast cancer in Chinese women younger than age 40 and a comparison with their older counterparts. MATERIALS AND METHODS: Demographic and clinicopathologic findings of 1,485 Chinese women with breast cancer seen during the period September 2003 to November 2006 were prospectively recorded, and comparisons were made between those who were under the age of 40 and those 40 years of age and older. These results were then compared with a reference population obtained from the Surveillance, Epidemiology and End Results (SEER) database between 2003 and 2004. RESULTS: 17.6% of the women were younger than 40 years old, and age distribution was significantly different from women in the SEER database. The mean age at menarche was lower in women under age 40 (p < 0.0005), and age at first live birth was also higher (p = 0.017). The rate of first detection by screening mammography was significantly higher among women who were 40 of age and older (p = 0.002). Breast conservation surgery was more commonly performed in the younger age group of Chinese women, particularly when tumor size was less than 2 cm (p = 0.001). A significantly higher proportion of women under age 40 had breast reconstruction (p < 0.001). The majority of women presented with stage 0-II disease, but in the Chinese groups the younger patients presented at a later stage (p = 0.04). Younger women had higher pathological grade and poorly differentiated tumors (p = 0.02), more nodal involvement (p = 0.024), and lymphovascular permeation involvement (p < 0.001). The majority of tumors were ER and PR positive in both groups, but younger women had a higher proportion of cerbB2-positive tumors. CONCLUSION: Chinese women present with breast cancer at an earlier age. Younger women present with more advanced disease and more aggressive tumor characteristics. More ethnic-specific screening protocols and treatment decisions may benefit this group of patients. PMID- 18408961 TI - Diagnostic accuracy of CT and ultrasonography for evaluating metastatic cervical lymph nodes in patients with thyroid cancer. AB - BACKGROUND: The present study was designed to investigate the diagnostic ability of computed tomography (CT) and ultrasonography (USG) in the preoperative evaluation of the cervical nodal status of patients with thyroid cancer. METHODS: The study population consisted of 37 consecutive patients (female:male = 30:7, age range: 20-68 years) who subsequently underwent total thyroidectomy and neck dissection for thyroid cancer. The results of the review of the preoperative CT and those of the original USG reports were compared with the histopathologic results. The accuracy was evaluated by "per level" and "per patient" analyses of whether the CT or USG results had or had not altered the choice of surgical method. RESULTS: By "per level" analysis, the sensitivities, specificities, and diagnostic accuracies were 77%, 70%, 74% for CT and 62%, 79%, 68% for USG, respectively, with a significant difference in the sensitivities (p = 0.002). When the lymph node levels were grouped into central and lateral compartments, all of the values for the lateral compartment tended to be higher than those for the central compartment for both CT (78%, 78%, 78% versus 74%, 44%, 64%) and USG (65%, 82%, 71 versus 55%, 69%, 60%). By per patient analysis, the sensitivities, specificities, and diagnostic accuracies of CT and USG were 100%, 90%, 97% and 100%, 80%, 95%, respectively. CONCLUSION: Despite of very high accuracy of USG by per patient analysis, the superior sensitivity of CT on the per level analysis may enable CT to play a complementary role for determining the surgical extent in selected patients with thyroid cancer. PMID- 18408962 TI - Choledochal cysts in children: epidemiology and outcomes. AB - BACKGROUND: The purpose of the present study was to evaluate the epidemiology and outcomes of choledochal cysts in children. METHODS: We performed a retrospective review of the records of all cases of choledochal cyst in children 0-14 years of age presenting at Chiang Mai University Hospital from May 2000 to February 2007. Demographic and clinical data, including laboratory and radiographic studies, as well as surgical and nonsurgical treatments and outcomes, were recorded. RESULTS: There were 32 patients (25 female) with a mean age at diagnosis of 4.1 years (range: 1 month to 14.8 years). The most common clinical presentation was jaundice (n = 17), followed by abdominal pain (n = 16), nausea/vomiting (n = 10), and abdominal mass (n = 8). Based on the Tondani modification of the Alonso-Lej classification, 20 cases were type I, whereas 9, 2, and 1 of the patients had types IV, V, and II, respectively. Biliary tract infections, including cholecystitis (n = 5) and cholangitis (n = 3), were the most common preoperative complications. Twenty-six patients had definitive surgery consisting of cystectomy with Roux-en-Y hepaticojejunostomy. Postoperative complications were noted in 3 patients and included intestinal obstruction, bleeding, and cholangitis. Three patients died, all younger than 2 years of age (p = 0.022). CONCLUSION: The epidemiology of choledochal cysts in Thai children was similar to cases reported from other regions. Early diagnosis and treatment, particularly in children under 2 years of age, should be emphasized, because these patients are at a higher risk for development of chronic liver disease and poor outcome. PMID- 18408963 TI - Surgeons underestimate their patients' desire for preoperative information. AB - BACKGROUND: Provision of adequate patient information may contribute to a "satisfying" surgical treatment. The patient's views on successful transfer of information concerning operative characteristics may not be in concert with the surgeon's. The aim of the present study was to determine opinions of both surgeons and patients about issues of surgical information. METHODS: A group of surgeons (n = 24) and surgical patients (n = 125) responded to a questionnaire that included 80 topics involving domains of information on disease, physical examination, preoperative period, anesthesia, operation, postoperative period, self care, and general hospital issues. Both groups were asked for their opinion on what they considered important and useful preoperative information for patients. Questions were scored with a visual analog scale. The reliability of the questionnaire was calculated with Cronbach's alpha. Differences in opinions between surgeons and patients were analyzed with Student's t-test. RESULTS: The Cronbach's alpha of the questionnaire was high (0.91), indicating its high reliability. Patients scored significantly higher (p < 0.001) in most domains, including preoperative period, anaesthesia, operation, postoperative period, self care, and general hospital information. Women demonstrated a significantly higher need for information than men did. These findings were independent of patient age or complexity of operation. In contrast, surgeons thought that their patients desired more extensive information on cause, effect, and prognosis of the disease itself (p < 0.001). CONCLUSION: Surgeons generally underestimate their patients' desire for receiving extensive information prior to a surgical procedure of any complexity. Surgeons should develop strategies to bridge this informational mismatch. PMID- 18408964 TI - Symposium on evidence-based endocrine surgery (2): benign thyroid disease. PMID- 18408965 TI - Fine-needle aspiration biopsy of benign thyroid nodules: an evidence-based review. AB - BACKGROUND: Since the development of fine-needle aspiration biopsy (FNAB) techniques, preoperative diagnosis and subsequent strategies for patient treatment have changed and evolved greatly. This is true also for thyroid FNAB: the vast majority of thyroid nodules are benign, and hence do not necessarily require surgical treatment. METHODS: A comprehensive Medline and Cochrane Library search was performed evaluating FNAB in the thyroid. In the last decade more than 400 articles on the subject have been published. Data in relation to the experience with FNAB at the Karolinska University Hospital since its introduction were also reviewed. RESULTS: The development of FNAB since the 1960s at the Karolinska University Hospital is described. During the period 1992-1996 the accuracy of the clinical routine was evaluated by studying the outcomes of almost 4,000 FNAs of the thyroid. The results were good, with only a few false-negative and false-positive results, but the problem of differentiating follicular adenoma from follicular carcinoma remained a significant problem. The use of immunological analysis has greatly increased the possibility of obtaining valuable information on cellular characteristics. CONCLUSION: A successful FNAB service rests on several factors, and the importance of clinical conferences between all specialists involved in the diagnosis and treatment of patients with thyroid disorders cannot be overemphasized. At the Karolinska University Hospital there are weekly conferences where patients are discussed both pre- and postoperatively. These conferences lead to optimal interaction between the different specialists and, most important, substantial improvement in the clinical management of patients with thyroid disorders. PMID- 18408966 TI - Acute care surgery: a new training and practice model in the United States. AB - The specialty of trauma is at a crossroads. Choosing a career in trauma is associated with concerns related to lifestyle issues and maintenance of adequate operative experience. Trauma and critical care surgeons in the U.S. have reexamined their role based on these concerns and the realization that surgeon resources for the injured patient are in jeopardy. After much work over the past five years, a model of "Acute Care Surgery" has emerged and a training curriculum has been proposed. This article reviews the evolution of a new specialty and identifies some of the challenges and opportunities associated with the implementation of this model. PMID- 18408967 TI - Acinetobacter baumannii infections in a surgical intensive care unit: predictors of multi-drug resistance. AB - OBJECTIVES: This study was designed to evaluate Acinetobacter baumannii infections incidence in our Surgical Intensive Care Unit, clinical features and outcome of these patients, and multi-resistance incidence to identify predictors of such a resistance. METHODS: Prospective study of all patients with ICU acquired Acinetobacter baumannii infection from June 1, 2003 to May 31, 2005. Patients with multi-resistant infection, susceptible exclusively to colistin, were compared with those sustaining non-multi-drug resistant infection. RESULTS: Among 411 patients, 52 (12.6%) developed Acinetobacter infection. Their mean age was 66.3 +/- 8.4 years and APACHE II 20.4 +/- 7.3 (men: 51.9%). Infection sites were: bloodstream (46.2%), respiratory tract (32.7%), central venous catheter (11.5%), surgical site (7.7%), and urinary tract (1.9%). High multi-resistance (44.2%), morbidity (63.4%), and mortality (44.2%) were identified. Colistin was the most effective antibiotic (100% susceptibility), whereas resistance against all other antibiotics was >60%. Previous septic shock (p = 0.04), previous adult respiratory distress syndrome (ARDS) (p = 0.01), number of previous antibiotics (p = 0.01), previous aminoglycoside use (p = 0.04), and reoperation (p = 0.01) were risk factors for multi-resistance in univariate analysis. Morbidity in the multi-resistant group was significantly higher than the non-multi-resistant group (82.6% vs. 48.2%, p = 0.02). Mortality in the multi-resistant group also was higher; however, this difference did not marginally reach statistical significance (60.8% vs. 31.1%, p = 0.06). Multivariate analysis identified previous septic shock (p = 0.04; odds ratio (OR), 9.83; 95% confidence interval (CI), 1.003-96.29) and reoperation (p = 0.01; OR, 8.45; 95% CI, 1.52-46.85) as independent predictors of multi-resistance. CONCLUSION: Acinetobacter baumannii infections are frequent and associated with high morbidity, mortality, and multi resistance. Avoidance of unnecessary antibiotics is a high priority, and specific attention should be paid to patients with previous ARDS and, particularly, previous septic shock and reoperation. When such risk factors are identified, colistin may be the only appropriate treatment. PMID- 18408968 TI - Effects of 3-week outpatient pulmonary rehabilitation on exercise capacity, dyspnea, and quality of life in COPD. AB - The effects of intensive 3-week outpatient pulmonary rehabilitation (PR) on exercise capacity, dyspnea, and health-related quality of life (HRQL) were investigated in patients with COPD. Two hundred ten patients with COPD (mean FEV(1) = 54%pred) underwent PR consisting of exercise training, patient and psychosocial education, breathing and relaxation therapy, nutrition counseling, and smoking cessation support. Before and after PR, exercise capacity was assessed with 6-min walking tests (6MWT) and constant cycle ergometer exercise (CEE). Dyspnea was measured after 6MWTs with a Borg scale and after PR with the Transition Dyspnoea Index (TDI). HRQL was examined with the Medical Outcomes Study Short Form 36 (SF-36). Results showed improvements in the 6MWT (+39 m, p < 0.001) and CEE (+241 W x min, p < 0.001) after PR, paralleled by decreased dyspnea during the 6MWT (-0.5, p < 0.001) and during activities (TDI score = 3.6). Increases in all SF-36 subscales reflected improved HRQL after PR (p < 0.001). No gender differences were found. Patients with milder versus more severe COPD improved similarly in most outcomes. Regression analyses revealed that TDI scores were the most important predictor of improvements in HRQL. The results suggest that intensive 3-week outpatient PR is associated with improvements in exercise capacity, dyspnea, and HRQL in male and female patients with COPD irrespective of COPD severity. Reduced dyspnea during activities contributed the most to improvements in HRQL. PMID- 18408969 TI - Lymphangioleiomyomatosis and tuberous sclerosis complex. AB - Lymphangioleiomyomatosis (LAM) is a rare multisystemic disease of women of child bearing age and affects mainly the lungs, promoting cystic destruction of lung parenchyma or leading to abdominal tumor formation (e.g., angiomyolipomas, lymphangioleiomyomas). LAM can arise sporadically or in association with tuberous sclerosis complex (TSC), an autosomal inherited syndrome characterized by hamartoma-like tumor growth and pathologic features that are distinct from manifestations of pulmonary LAM. A substantial body of evidence has now been gathered suggesting that the two diseases share a common genetic origin. TSC is caused by mutations in two genes, TSC1 on chromosome 9q34 and TSC2 on 16p13. Both of these genes are tumor suppressor genes encoding hamartin (TSC1) and tuberin (TSC2). Sporadic LAM is correlated with a mutation in the TSC2 gene and tuberin appears to play a central role in the pathogenesis of the disease. A TSC2 loss or mutation leads to disruption of the tuberin-hamartin heteromer and dysregulation of S6K1 activation leading to aberrant cell proliferation seen in LAM disease. The extremely diverse clinical and radiologic features of the disease and the complex therapeutic approach are reviewed in detail. Although new therapeutic agents have been tested, to date no effective treatment has been proposed and the prognosis of patients with LAM remains poor. As long as newer therapeutic agents do not change this picture, lung transplantation remains the last hope for patients with respiratory failure at the advanced stage of the disease. PMID- 18408971 TI - Risk factors and oncologic impact of anastomotic leakage after rectal cancer surgery. AB - PURPOSE: The impact of anastomotic leakage on the long-term oncologic outcome is not clear. This retrospective study evaluated risk factors and oncologic impacts of anastomotic leakage after rectal cancer surgery. METHODS: Data were analyzed from 1,391 patients who underwent sphincter preservation for rectal cancer between January 1997 and August 2003. Operations were classified as anterior resection (n = 164), low anterior resection (n = 898), or ultralow anterior resection (n = 329). RESULTS: The anastomotic leakage rate was 2.5 percent. Multivariate analysis identified male (hazard ratio, 3.03), old age (hazard ratio, 2.42), and lower anastomosis level (hazard ratio, 2.68) as risk factors for leakage. The local recurrence rates were 9.6 and 2.2 percent for the leakage and nonleakage groups, respectively but were not significant (P = 0.14). The overall five-year survival rates were 55.1 and 74.1 percent in the leakage and nonleakage groups, respectively (P < 0.05), and the cancer-specific survival rates were 63 and 78.3 percent in the leakage and nonleakage groups, respectively (P = 0.05). However, in subgroup analysis, significant differences were identified only in Stage III patients. CONCLUSIONS: Age, sex, and ultralow anterior resection were found to be risk factors for anastomotic leakage after rectal cancer surgery. In addition, leakage was associated with poor survival. PMID- 18408970 TI - Using the zebrafish lateral line to screen for ototoxicity. AB - The zebrafish is a valuable model for studying hair cell development, structure, genetics, and behavior. Zebrafish and other aquatic vertebrates have hair cells on their body surface organized into a sensory system called the lateral line. These hair cells are highly accessible and easily visualized using fluorescent dyes. Morphological and functional similarities to mammalian hair cells of the inner ear make the zebrafish a powerful preparation for studying hair cell toxicity. The ototoxic potential of drugs has historically been uncovered by anecdotal reports that have led to more formal investigation. Currently, no standard screen for ototoxicity exists in drug development. Thus, for the vast majority of Food and Drug Association (FDA)-approved drugs, the ototoxic potential remains unknown. In this study, we used 5-day-old zebrafish larvae to screen a library of 1,040 FDA-approved drugs and bioactives (NINDS Custom Collection II) for ototoxic effects in hair cells of the lateral line. Hair cell nuclei were selectively labeled using a fluorescent vital dye. For the initial screen, fish were exposed to drugs from the library at a 100-muM concentration for 1 h in 96-well tissue culture plates. Hair cell viability was assessed in vivo using fluorescence microscopy. One thousand forty drugs were rapidly screened for ototoxic effects. Seven known ototoxic drugs included in the library, including neomycin and cisplatin, were positively identified using these methods, as proof of concept. Fourteen compounds without previously known ototoxicity were discovered to be selectively toxic to hair cells. Dose-response curves for all 21 ototoxic compounds were determined by quantifying hair cell survival as a function of drug concentration. Dose-response relationships in the mammalian inner ear for two of the compounds without known ototoxicity, pentamidine isethionate and propantheline bromide, were then examined using in vitro preparations of the adult mouse utricle. Significant dose-dependent hair cell loss in the mouse utricle was demonstrated for both compounds. This study represents an important step in validating the use of the zebrafish lateral line as a screening tool for the identification of potentially ototoxic drugs. PMID- 18408972 TI - Massive lower gastrointestinal hemorrhage caused by a large extraluminal leiomyoma of the colon: report of a case. AB - The occurrence of leiomyoma of the colon is uncommon. Most of these lesions are clinically silent and are found incidentally during laparotomy or endoscopic procedures for unrelated conditions. Symptomatic leiomyomas of the colon are encountered less frequently, with only sporadic reports in the literatures. We describe a heretofore unreported case of a large extraluminal leiomyoma of the sigmoid colon presenting as massive lower gastrointestinal hemorrhage. Because it was extraluminal in position, it was difficult to make an accurate diagnosis endoscopically and the condition was easily misdiagnosed as angiodysplasia of the colon until CT scan results were seen. Although rare and benign in nature, leiomyoma of the colon may cause life-threatening complications that require emergency surgery and should be included in the differential diagnosis of lower gastrointestinal hemorrhage. PMID- 18408973 TI - Defense-inducing volatiles: in search of the active motif. AB - Herbivore-induced volatile organic compounds (VOCs) are widely appreciated as an indirect defense mechanism since carnivorous arthropods use VOCs as cues for host localization and then attack herbivores. Another function of VOCs is plant-plant signaling. That VOCs elicit defensive responses in neighboring plants has been reported from various species, and different compounds have been found to be active. In order to search for a structural motif that characterizes active VOCs, we used lima bean (Phaseolus lunatus), which responds to VOCs released from damaged plants with an increased secretion of extrafloral nectar (EFN). We exposed lima bean to (Z)-3-hexenyl acetate, a substance naturally released from damaged lima bean and known to induce EFN secretion, and to several structurally related compounds. (E)-3-hexenyl acetate, (E)-2-hexenyl acetate, 5-hexenyl acetate, (Z)-3-hexenylisovalerate, and (Z)-3-hexenylbutyrate all elicited significant increases in EFN secretion, demonstrating that neither the (Z) configuration nor the position of the double-bond nor the size of the acid moiety are critical for the EFN-inducing effect. Our result is not consistent with previous concepts that postulate reactive electrophile species (Michael-acceptor systems) for defense-induction in Arabidopsis. Instead, we postulate that physicochemical processes, including interactions with odorant binding proteins and resulting in changes in transmembrane potentials, can underlie VOCs-mediated signaling processes. PMID- 18408974 TI - Compound muscle action potentials and spontaneous electromyography can be used to identify and protect the femoral nerve during resection of large retroperitoneal tumors. AB - BACKGROUND: Resection of large retroperitoneal neoplasms may injure the femoral nerve, thereby causing a permanent neurological deficit. We used electrical neurophysiological monitoring to identify, map, and preserve the femoral nerve during surgical resection to reduce the risk of neurological deficit. METHODS: Seven patients with retroperitoneal neoplasms underwent eight resections. Compound muscle action potentials (CMAPs) were recorded from needle electrodes placed in the iliacus, quadriceps, and sartorius muscles. Spontaneous electromyography (EMG) was continuously monitored from the same muscle groups. A handheld monopolar stimulator was used to elicit evoked EMG responses to identify and map the course of the femoral nerve. A stimulating strength of 10 mA was used to map the nerve. The stimulation threshold was tested after neoplasm resection to predict postoperative femoral nerve function. RESULTS: Electrical stimulation with CMAP recording and a stimulating strength of 10 mA successfully localized the femoral nerve in six cases. Monitoring with a stimulating threshold between 0.6 and 1.6 mA predicted postoperative femoral nerve preservation after tumor resection in four of the six cases. CONCLUSION: Neurophysiological monitoring using CMAP and spontaneous EMG can protect the femoral nerve during resection of large retroperitoneal neoplasms. PMID- 18408975 TI - Multivariate analysis of the pathologic features of esophageal squamous cell cancer: tumor budding is a significant independent prognostic factor. AB - BACKGROUND: Tumor budding has been suggested to be a prognostic factor in various cancers but has never been studied in esophageal cancer. METHODS: In this study, the microscopic finding of tumor budding in esophageal squamous cell carcinoma was correlated with outcome after esophagectomy. One hundred and thirty-six patients undergoing a curative esophagectomy were assigned to either a frequent (n = 82) or rare (n = 54) group according to the microscopically observed frequency of tumor budding in the tumor. RESULTS: The 5-year survival rates after esophagectomy were 35.4% for the frequent group and 81.3% for the rare group. Multivariate analysis using the Cox proportional hazards model by a stepwise method identified this morphological variable as a significant independent prognostic factor. CONCLUSIONS: Tumor budding in esophageal squamous cell carcinoma reflects the biological activity of the tumor and may be a useful prognostic indicator. PMID- 18408976 TI - Poor hormone receptor expression in East African breast cancer: evidence of a biologically different disease? AB - BACKGROUND: Few studies have examined breast cancer hormone receptor expression in Africans. We report on the hormone receptor profile of breast cancer in East Africans in the largest prospective study for this region. METHODS: Consecutive breast cancer presentations to a hospital in Kijabe (2001-2007) were included. Demographic, clinical, and test data were collected. ER/PR and Her2 testing was based on immunohistochemistry (IHC). RESULTS: There were 129 subjects (median 47 years), most had invasive ductal cancer and locally advanced disease and/or metastases. ER/PR testing was done in 120: 24% had ER-positive tumours, 34% were ER- and/or PR-positive, 10% were ER-negative but PR-positive tumours, and 66% were negative for ER and PR. ER/PR positivity was not associated with stage (P = 0.28) and was not related to age, parity, menopausal status, or node metastases. Increasing tumour grade was associated with PR expression (P = 0.02) with decreasing frequency of PR positive tumours as histological grade increased; there was weak evidence of an association between grade and ER expression (P = 0.06). Of cases tested, 26.5% overexpressed Her2. CONCLUSIONS: Breast cancer in Kijabe is an advanced-stage disease, comprised mainly of poorly differentiated cancers that are less likely to be hormone sensitive (across all stages of disease). ER/PR testing of all those affected by breast cancer should be supported as a global priority in cancer control. International and inter-African research collaborations are needed to allow genetic detailing of tumours in indigenous Africans to assess possible racial heterogeneity in the biology of breast cancer. PMID- 18408977 TI - Reproducibility of semi-quantitative parameters in FDG-PET using two different PET scanners: influence of attenuation correction method and examination interval. AB - PURPOSE: The aim of this study is to evaluate the reproducibility of semi quantitative parameters obtained from two 2-deoxy-2-[F-18]fluoro-D-glucose positron emission tomography (FDG-PET) studies using two different PET scanners. METHODS: Forty-five patients underwent FDG-PET examination with two different PET scanners on separate days. Two PET images with different attenuation correction method were generated in each patient, and three regions of interest (ROIs) were placed on the lung tumor and normal organs (mediastinum and liver) in each image. Mean and maximum standardized uptake values (SUVs), tumor-to-mediastinum and tumor-to-liver ratios (T/M and T/L), and the percentage difference in parameters between two PET images (% Diff.) were compared. RESULTS: All measured values except maximum SUV in the liver and tumor-related parameters (SUV in lung tumor, T/M, T/L) showed no significant difference between two PET images. CONCLUSION: The mean measured values showed high reproducibility and demonstrate that follow up study or measurement of tumor response to anticancer drugs can be undertaken by FDG-PET examination without specifying the particular type of PET scanner. PMID- 18408978 TI - Launois-Bensaude Syndrome: an unusual localization of obesity disease. AB - BACKGROUND: Launois-Bensaude syndrome is a rare pathology consisting of adipose masses symmetrically distributed mainly in the superior part of the body. Men are especially affected between age of 30 and 60 as well as chronic alcohol abusers. Etiopathogenesis is attributable to mutations or deletions of mitochondrial DNA, and alcohol is a possible cofactor. METHODS: The current treatment of the disease is described based on the authors' experience. Four cases treated in our department are retrospectively reviewed regarding comorbidities and type of surgery performed. RESULTS: A relevant and long-lasting reduction of fat bulges has been obtained in all cases with no major complications except for a mild anemia. CONCLUSION: Launois-Bensaude syndrome causes a functional rather than esthetic concern due to the peculiar localization of fat bulges. Currently, the only effective therapy is surgery, through lipectomy or liposuction of adipose bulges. PMID- 18408979 TI - Laparoscopic gastric rebanding for slippage with pouch dilation: results on 29 consecutive patients. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) slippage with pouch dilation is one the most serious long-term complications and requires reoperation in most cases. It is still controversial whether banding should be offered again or a different procedure should be chosen. We report the results of synchronous de-rebanding on a prospective series of patients treated at our institution for slippage with pouch dilation. METHODS: From January 2000 to May 2007, 29 consecutive patients underwent laparoscopic de-rebanding for slippage with pouch dilation. The mean age at primary operation was 38.9 +/- 8.2 years and the mean BMI was 46.4 +/- 8 kg/m(2). Twenty-eight had previous LAGB, while one had previous open gastric banding, the perigastric technique being used at that time. All the redo procedures were successfully carried out under laparoscopy, via the pars flaccida technique, and all the patients were followed-up according to the usual schedule. RESULTS: The mean time from the original LAGB was 45.3 +/- 30.3 months, and the mean follow-up after rebanding was 26.9 +/- 20.6 months. At rebanding, the mean BMI was 34.3 +/- 7.6, percent excess weight loss (%EWL) 54.5 +/- 31, and percent excess BMI loss (%EBL) 58.3 +/- 33, respectively. After 1 year, BMI was 36.3 +/- 7.9, %EWL 40.8 +/- 30.5, and %EBL 43.9 +/- 32.7, respectively. After 2 years, BMI was 37.13 +/- 7.4, %EWL 36.9 +/- 29.4, and %EBL 39.6 +/- 31.6, respectively, and after 3 years, BMI was 33.5 +/- 5.6, %EWL 51.9 +/- 24.3, and %EBL 55.7 +/- 25.7, respectively. One patient had re-rebanding after 6 months for a new slippage, two had band removal with refusal to switch to another procedure, one had biliopancreatic diversion for slippage recurrence, and one underwent sleeve gastrectomy for insufficient weight loss after 6 months. CONCLUSIONS: Although this is a limited series, our results show that good outcomes can be expected after rebanding in properly assessed patients with slippage and pouch dilation. Larger series and longer follow-up are needed to confirm these findings. PMID- 18408981 TI - Weight loss reporting. PMID- 18408980 TI - The effect of laparoscopic gastric banding surgery on plasma levels of appetite control, insulinotropic, and digestive hormones. AB - BACKGROUND: We hypothesized that laparoscopic adjustable gastric banding (LAGB) reduces weight and modulates ghrelin production, but largely spares gastrointestinal endocrine function. To examine this hypothesis, we determined plasma concentrations of appetite-control, insulinotropic, and digestive hormones in relation to LAGB. METHODS: Twenty-four patients undergoing LAGB were prospectively enrolled. Body mass index (BMI) was measured and blood samples obtained at baseline and 6 and 12 months post-surgery. Plasma concentrations of leptin, acylated and total ghrelin, pancreatic polypeptide (PP), insulin, glucose dependent insulinotropic peptide (GIP), active glucagon-like peptide-1 (GLP-1), gastrin, and pepsinogens I and II were measured using enzyme-linked immunoassays. RESULTS: Median percent excess weight loss (%EWL) over 12 months was 45.7% with median BMI decreasing from 43.2 at baseline to 33.8 at 12 months post-surgery (p < 0.001). Median leptin levels decreased from 19.7 ng/ml at baseline to 6.9 ng/ml at 12 months post-surgery (p < 0.001). In contrast, plasma levels of acylated and total ghrelin, PP, insulin, GIP, GLP-1, gastrin, and pepsinogen I did not change in relation to surgery (p > 0.05). Pepsinogen II levels were significantly lower 6 months after LAGB but returned to baseline levels by 12 months. CONCLUSIONS: LAGB yielded substantial %EWL and a proportional decrease in plasma leptin. Our results support the hypothesis that LAGB works in part by suppressing the rise in ghrelin that normally accompanies weight loss. Unchanged concentrations of insulinotropic and digestive hormones suggest that gastrointestinal endocrine function is largely maintained in the long term. PMID- 18408982 TI - Hunger control and regular physical activity facilitate weight loss after laparoscopic adjustable gastric banding. AB - BACKGROUND: Bariatric surgery facilitates substantial and durable weight loss; however, outcomes vary. In addition to physiological and technical factors, weight loss efficacy is dependent on modification of behavior to maintain a long term change in energy balance. This study aimed to assess the extent and nature of change in energy intake and physical activity and identify factors associated with percentage weight loss (%WL) 12 months after laparoscopic adjustable gastric banding (LAGB). METHODS: 129 bariatric surgery candidates (26 men/103 women, mean age 45.2 +/- 11.5, mean body mass index [BMI] 44.3 +/- 6.8, range 31.9 to 66.7) completed the study. Data were collected at baseline and 12 months. Validated questionnaires included the Cancer Council Victoria Food Frequency Questionnaire, Three Factor Eating Questionnaire, Short Form-36, Baecke Physical Activity Questionnaire, and Beck Depression Inventory. Symptoms of "non-hungry eating," "emotional eating," and "grazing" were assessed. RESULTS: Mean %WL was 20.8 +/- 8.5%, and excess weight loss was 50.0 +/- 20.7 (p < 0.001). Mean total energy intake reduced from 9,991 +/- 3,986 kj to 4,077 +/- 1,493 kj (p < 0.001). Average leisure time and sport-related physical activity scores increased (both p < 0.001). Regression analysis identified baseline BMI (beta = 0.241; p = 0.002), subjective hunger (beta = -0.275; p = 0.001), physical function (beta = 0.309; p < 0.001), and leisure time physical activity (beta = 0.213; p = 0.010) as independent predictors of %WL, total R (2) 0.34%. "Non-hungry eating" and symptoms of depression were also related to poorer %WL. CONCLUSION: LAGB affects marked behavior change and facilitates substantial weight loss in the first 12 months. However, variations in adopted behaviors can affect energy balance and weight loss success. Achievement and maintenance of favorable behaviors should be an important consideration during on-going postsurgical review and counseling. Management should include adequate band adjustment to control physical hunger, optimization of physical function and activity, and reinforcement of strategies to reduce energy intake. PMID- 18408983 TI - Laparoscopic Roux-en-Y gastric bypass in morbidly obese patients > or =55 years old. AB - BACKGROUND: Advanced age is considered a relative contraindication to bariatric surgery at some institutions because of concerns about higher morbidity and less than optimal weight loss. The aim of our study was to evaluate the operative outcomes, length of stay, weight loss, and improvement of comorbidities in patients > or =55 years old who underwent laparoscopic Roux-en-Y gastric bypass (LRYGB) surgery in our institution. METHODS: Retrospective data on 33 patients (26 women and 7 men) > or =55 years of age who underwent LRYGB from January 2003 to December 2006 were reviewed. RESULTS: Average patient age was 59 years (range 55-68 years), and the mean preoperative body mass index was 47 kg/m2 (range 41.1 55.8 kg/m2). The median length of hospital stay was 3 days. There were no intraoperative or postoperative deaths. Early complications were one anastomotic leak, two upper gastrointestinal bleedings, and two readmissions for intractable vomiting. Late complications included four anastomotic strictures and one small bowel obstruction. Patients were followed for a mean 13 months (range 3-24 months). The mean excess body weight (EBW) loss was 13.5 kg (23%), 23.3 kg (39.8%), 33.3 kg (58.1%), 39.8 kg (66.8%), 40.1 kg (69.5%), and 40.8 kg (75.3%) at 1, 3, 6, 9, 12 and 24 months, respectively. Diabetes mellitus improved in 19 (100%) patients and completely resolved in 10 (53%). Hypertension improved in 18 (64%) patients, completely resolved in 9 (32%) and was unchanged in 10 (36%). CONCLUSIONS: LRYGB is safe and effective in morbidly obese patients > or =55 years of age. PMID- 18408984 TI - Laparoscopic totally stapled entero-enterostomy: a safe and reproducible new technique. AB - BACKGROUND: Laparoscopic gastric bypass for morbid obesity is a technically demanding procedure, partially because of the necessity to construct two anastomoses. In this study, a new technique to perform the entero-enterostomy is presented. METHODS: We evaluated the procedure in a consecutive series of 100 patients who underwent laparoscopic gastric bypass. Intra- and postoperative complications were analyzed. RESULTS: No complications in relation to the construction of the entero-enterostomy occurred. No leakage, kinking, or stenosis were observed during a mean follow-up of 13.5 months (range 6-20 months). CONCLUSION: This new technique for a totally stapled entero-enterostomy can be recommended. PMID- 18408985 TI - Possible roles of mast cell-derived chymase for skin rejuvenation. AB - The relationships between mast cell-derived chymase, angiotensin II, and extracellular matrix production in the skin after intense pulsed light (IPL) were clarified in hamsters. Dorsal areas of the hamsters were irradiated once or twice a week by IPL. The index of extracellular matrix production in the skin was defined as the depth stained with Azan-Mallory stain from the epidermis to the dermis at the point of maximum thickness. The index had significantly increased 7 days after IPL irradiation in sections treated once or twice with IPL compared with that of untreated control sections. The numbers of mast cells, chymase positive cells, and angiotensin II-positive cells had also significantly increased in IPL-irradiated areas. Significant increases in chymase and angiotensin II activities were observed in the extracts obtained from IPL irradiated skin. Mast cell-derived chymase may be involved via angiotensin II formation in the dermal extracellular matrix production that occurs after IPL irradiation. PMID- 18408986 TI - Relieving pain in minor aphthous stomatitis by a single session of non-thermal carbon dioxide laser irradiation. AB - This randomized controlled clinical trial was designed to evaluate the efficacy of single-session, non-thermal, carbon dioxide (CO(2)) laser irradiation in relieving the pain of minor recurrent aphthous stomatitis (miRAS) as a prototype of painful oral ulcers. Fifteen patients, each with two discrete aphthous ulcers, were included. One of the ulcers was randomly allocated to be treated with CO(2) laser (1 W of power in de-focused continuous mode) and the other one served as a placebo. Before laser irradiation, a layer of transparent, non-anesthetic gel was placed on both the laser lesions and the placebo lesions. The patients were requested to grade their pain on a visual analog scale up to 96 h post operatively. The reduction in pain scores was significantly greater in the laser group than in the placebo group. The procedure itself was not painful, so anesthesia was not required. Powermetry revealed the CO(2) laser power to be 2-5 mW after passing through the gel, which caused no significant temperature rise or any visual effect of damage to the oral mucosa. Our results showed that a low intensity, non-thermal, single-session of CO(2) laser irradiation reduced pain in miRAS immediately and dramatically, with no visible side effects. PMID- 18408989 TI - Aceruloplasminemia: a case report. PMID- 18408990 TI - Protein structure and function: structural biology, bioinformatics, and system biology. Editorial. PMID- 18408991 TI - The Childhood Autism Spectrum Test (CAST): sex differences. AB - The Childhood Autism Spectrum Test (CAST) (formally known as the Childhood Asperger Screening Test) identifies autism spectrum conditions by measuring social and communication skills. The present study explored the sex distribution of scores. The CAST was distributed to 11,635 children aged 4-9 years in Cambridgeshire primary schools (UK). 3,370 (29%) were returned. The median score was significantly higher in boys (median test, P < 0.001) (Boys, median = 5 (IQR: 3,8); girls, median = 4 (IQR: 2,6)). There was a predominance of boys (n = 81 (79.4%) over girls (n = 21 (20.6%)) amongst those scoring >or= 15. Exclusion of children with ASC did not significantly affect the results. We conclude that different profiles of social and communication skills in boys and girls must be taken into account when measuring these skills in the general population. PMID- 18408992 TI - No correlation between heterozygosity and vegetative fitness in the narrow endemic and critically endangered Clematis acerifolia (Ranunculaceae). AB - The relationship between heretozygosity and vegetative fitness was explored in the narrow endemic and threatened Clematis acerifolia (Ranunculaceae), both at individual and population levels. The relationships between fitness, habitat factors, and population size were also analyzed. Allozyme electrophoresis was used to quantify the levels of heterozygosity of nearly 200 surveyed individuals belonging to the nine extant populations of this species. Six parameters of vegetative fitness were measured: plant height, shrub diameter, length of the largest leaf, width of the largest leaf, mean number of leaves/stem, and total number of stems. The percentage of tree cover (light availability) was measured as an indicator of habitat quality. A principal component analysis reduced the original fitness variables to two uncorrelated principal components. None of these correlated significantly with both heterozygosity and population size, in contrast to the expected result. Nevertheless, one of the principal components showed a positive relationship with light availability, which may indicate that habitat quality may have significant effects on the performance of this species. Thus, to ensure the viability of this endangered species, maintenance of adequate habitat quality (by avoiding further fragmentation) may be more important than maximizing genetic diversity within populations. PMID- 18408993 TI - The optimal negligence standard in health care under supply-side cost sharing. AB - This paper elaborates on the optimal negligence standard in a world where physicians choose their level of care subject to erroneous court judgements and to the degree of supply-side cost sharing. Uncertain liability in malpractice lawsuits leads physicians to provide excessive and insufficient care, which results in a loss of social welfare. The standard that maximizes welfare depends on the cost share: Under traditional, close to full cost reimbursement it is lower than the first-best level of care, while under substantial supply-side cost sharing it increases and may even exceed the first best. PMID- 18408994 TI - The ecotoxicology of nanoparticles and nanomaterials: current status, knowledge gaps, challenges, and future needs. AB - This paper introduces a special issue on the ecotoxicology and environmental chemistry of nanoparticles (NPs), and nanomaterials (NMs), in the journal Ecotoxicology. There are many types of NMs and the scientific community is making observations on NP ecotoxicity to inform the wider debate about the risks and benefits of these materials. Natural NPs have existed in the environment since the beginning of Earth's history, and natural sources can be found in volcanic dust, most natural waters, soils and sediments. Natural NPs are generated by a wide variety of geological and biological processes, and while there is evidence that some natural NPs can be toxic, organisms have also evolved in an environment containing natural NPs. There are concerns that natural nano-scale process could be influenced by the presence of pollution. Manufactured NPs show some complex colloid and aggregation chemistry, which is likely to be affected by particle shape, size, surface area and surface charge, as well as the adsorption properties of the material. Abiotic factors such as pH, ionic strength, water hardness and the presence of organic matter will alter aggregation chemistry; and are expected to influence toxicity. The physico-chemistry is essential to understanding of the fate and behaviour of NPs in the environment, as well as uptake and distribution within organisms, and the interactions of NPs with other pollutants. Data on biological effects show that NPs can be toxic to bacteria, algae, invertebrates and fish species, as well as mammals. However, much of the ecotoxicological data is limited to species used in regulatory testing and freshwater organism. Data on bacteria, terrestrial species, marine species and higher plants is particularly lacking. Detailed investigations of absorption, distribution, metabolism and excretion (ADME) remain to be performed on species from the major phyla, although there are some data on fish. The environmental risk assessment of NMs could be performed using the existing tiered approach and regulatory framework, but with modifications to methodology including chemical characterisation of the materials being used. There are many challenges ahead, and controversies (e.g., reference substances for ecotoxicology), but knowledge transfer from mammalian toxicology, colloid chemistry, as well as material and geological sciences, will enable ecotoxicology studies to move forward in this new multi-disciplinary field. PMID- 18408995 TI - Manufactured nanoparticles: their uptake and effects on fish--a mechanistic analysis. AB - There is an emerging literature reporting toxic effects of manufactured nanomaterials (NMs) and nanoparticles (NPs) in fish, but the mechanistic basis of both exposure and effect are poorly understood. This paper critically evaluates some of the founding assumptions in fish toxicology, and likely mechanisms of absorption, distribution, metabolism and excretion (ADME) of NPs in fish compared to other chemicals. Then, using a case study approach, the paper compares these assumptions for two different NPs; TiO2 and C60 fullerenes. Adsorption of NPs onto the gill surface will involve similar processes in the gill microenvironment and mucus layer to other substances, but the uptake mechanisms for NPs by epithelial cells are more likely to occur via vesicular processes (e.g., endocytosis) than uptake on membrane transporters or by diffusion through the cell membranes. Target organs may include the gills, gut, liver and sometimes the brain. Information on metabolism and excretion of NPs in fish is limited; but hepatic excretion into the bile seems a more likely mechanism, rather than mainly by renal or branchial excretion. TiO2 and C60 share some common chemical properties that appear to be associated with some similar toxic effects, but there are also differences, that highlight the notion that chemical reactivity can inform toxic effect of NPs in a fundamentally similar way to other chemicals. In this paper we identify many knowledge gaps including the lack of field observations on fish and other wildlife species for exposure and effects of manufactured NMs. Systematic studies of the abiotic factors that influence bioavailability, and investigation of the cell biology that informs on the mechanisms of metabolism and excretion of NMs, will greatly advance our understanding of the potential for adverse effects. There are also opportunities to apply existing tools and techniques to fundamental studies of fish toxicology with NPs, such as perfused organs and fish cell culture systems. PMID- 18408996 TI - Chronic, but not acute morphine treatment, up-regulates alpha-Ca2+/calmodulin dependent protein kinase II gene expression in rat brain. AB - The effects of acute and chronic morphine treatments on the expression of Ca2+/calmodulin dependent protein kinase II (CaMK II) gene in rat brain were investigated using in situ hybridization histochemistry. Our data showed that repeated, but not single morphine administration, resulted in significant up regulation of the alpha-CaMK II gene expression in hippocampus and frontal cortex. We further studied the time courses of alpha-CaMK II gene expression in response to repeated morphine administration. After 3 days of consecutive morphine injections, the alpha-CaMK II mRNA levels exhibited a trend of up regulation, and after 6 days of consecutive morphine injections it increased over 50-60% as compared with the control group. The alpha-CaMK II mRNA levels remained high 24 h after the cessation of chronic morphine treatment and returned to the control level 72 h later. However, changes of alpha-CaMK II gene levels mentioned above were not detected in amygdala or piriform cortex. Taken together, our data demonstrate that chronic morphine treatment region-specific up-regulates the levels of the alpha-CaMK II gene expression in hippocampus and frontal cortex. PMID- 18408997 TI - A hypothesis about the relationship of myelin-associated glycoprotein's function in myelinated axons to its capacity to inhibit neurite outgrowth. AB - The myelin-associated glycoprotein (MAG) is selectively localized in periaxonal Schwann cell and oligodendroglial membranes of myelin sheaths suggesting that it functions in glia-axon interactions in the PNS and CNS, and this is supported by much experimental evidence. In addition, MAG is now well known as one of several white matter inhibitors of neurite outgrowth in vitro and axonal regeneration in vivo, and this latter area of research has provided a substantial amount of information about neuronal receptors or receptor complexes for MAG. This article makes the hypothesis that the capacity of MAG to inhibit outgrowth of immature developing or regenerating neurites is an aberration of its normal physiological function to promote the maturation, maintenance, and survival of myelinated axons. The overview summarizes the literature on the function of MAG in PNS and CNS myelin sheaths and its role as an inhibitor of neurite outgrowth to put this hypothesis into perspective. Additional research is needed to determine if receptors and signaling systems similar to those responsible for MAG inhibition of neurite outgrowth also promote the maturation, maintenance, and survival of myelinated axons as hypothesized here, or if substantially different MAG-mediated signaling mechanisms are operative at the glia-axon junction. PMID- 18408998 TI - A tribute to Professor Ji-Sheng Han. PMID- 18409000 TI - Usefulness of multidetector computed tomography for detection of the obstructive point in a patient with ileus. PMID- 18408999 TI - Hypoxia ischemia-mediated cell death in neonatal rat brain. AB - The examination of Bcl-2-associated X protein (Bax) protein's role in the activation of cognate nuclear, mitochondrial and ER cell death signaling cascades and the resulting effects on cell death phenotype in the brain after neonatal hypoxia-ischemia (HI) requires an understanding of neonatal HI insult and progression, as well as, its dysfunctional outcomes. In addition, knowledge of key concepts of oxidative stress, a major injurious component of HI, and the different cell death phenotypes (i.e. apoptosis and necrosis) will aid the design of appropriate useful experimental paradigms. Here we discuss organelle cell death signaling cascades in the context of the different cell death phenotypes associated with animal models of neonatal hypoxia ischemia and tissue culture models used in the study of hypoxia ischemia, focusing on the intracellular shifts of the Bcl-2 associated X protein (Bax) in the hypoxic brain. PMID- 18409001 TI - Elevated HbA1c is an independent predictor of aggressive clinical behavior in patients with colorectal cancer: a case-control study. AB - AIM: The aim of this study was to seek an association between the control of type 2 diabetes mellitus (T2DM), as determined by hemoglobin A1c (HbA1c) levels, and the outcome of colorectal cancer (CRC). METHODS: We performed a retrospective review of patients with T2DM who had CRC diagnosed between 1997 and 2001. We defined well-controlled T2DM as HbA1c < 7.5% and poorly controlled T2DM as HbA1c > or = 7.5%. A group of age- and gender-matched patients who had CRC without T2DM were used as controls. Forty clinical factors were reviewed, and those associated with poor clinical outcome in each group were examined by univariate analysis (UA) and by the maximum likelihood analysis of logistic regression to determine the independent predictors of cancer outcome. RESULTS: We identified 155 patients with T2DM and CRC, and 114 control patients who had CRC without T2DM. We found no significant differences in any clinical factor by UA between the patients with well-controlled T2DM and the patients who had CRC without T2DM. Compared to both of those patients groups, in contrast, the patients with poorly controlled T2DM had more right-sided CRCs (P = 0.04, OR = 2, 95% CI = 1-4.1), more advanced CRCs (P = 0.02, OR = 2.1, 95% CI = 1-4.4), a younger age of presentation (P = 0.05), greater use of exogenous insulin (P = 0.002), and a poorer 5-year survival (P = 0.001) by UA. Logistic regression showed that poorly controlled T2DM independently predicted the early onset of CRC, a more advanced stage at the time of presentation, poorer 5-year survival, and an increased incidence of right sided CRCs. CONCLUSIONS: In patients with T2DM who have CRC, poor glycemic control is associated with a clinically aggressive course for the cancer. PMID- 18409002 TI - Correlation of quantitative assay of HBsAg and HBV DNA levels during chronic HBV treatment. AB - BACKGROUND AND AIM: Viral load is used for the diagnosis and monitoring the treatment of chronic hepatitis B (CHB). These methods are molecular-based and are expensive. Previous studies suggest that quantitative hepatitis B surface antigen (HBsAg) studied by automated chemiluminescent microparticle immunoassay can be a surrogate marker. In this study, we aimed to investigate whether quantitative HBsAg correlates hepatitis B virus (HBV) DNA levels during CHB treatment. METHODS: The study included 18 patients (13 male, 5 female, mean age: 33 +/- 9 years) with CHB. They were given pegylated interferon +/- lamivudine for 52 months and serum samples were obtained in weeks 0, 4, 8, 24, 48, 52, and 76. HBV DNA was measured by TaqMan polymerase chain reaction (PCR; Erasmus MC, University Medical Center, Rotterdam, The Netherlands). Quantitative HBsAg was studied by automated chemiluminescent microparticle immunoassay (Architect HBsAg, Abbott, IL). Results HBV DNA levels were measured as follows: 9.66, 7.69, 7.06, 5.93, 5.89, 5.88, and 7.27 logarithmic genome equivalent/ml, respectively. The corresponding HBsAg quantitation results were 42,888, 31,176, 37,882, 27,277, 28,279, 29,471, and 31,535 IU/ml, respectively. They showed a significant correlation (canonical correlation = 0.85). CONCLUSIONS: HBsAg studied by automated chemiluminescent microparticle immunoassay correlates with HBV DNA and can be a surrogate marker during the monitoring of the efficacy of HBV treatment. PMID- 18409003 TI - The effect of Helicobacter pylori eradication treatment on the MUC 1 and Lewis antigens level in human gastric juice: a preliminary study. AB - Helicobacter pylori is considered as a causative agent of gastritis, duodenal and gastric ulcers, and gastric cancer. During inflammation, association of the pathogen of gastric epithelial cells and mucins is considered important. It was postulated that Lewis b structures of secretory MUC 5AC mucin can be a receptor for the bacterium. Some authors also suggest that epithelial MUC 1 mucin may be implicated in the mechanism of infection. The main aim of our work was to support this last suggestion by evaluation of the possible changes in MUC 1 and Lewis a and b levels in gastric juice before and at the end of eradication treatment. The gastric juices of ten examined patients were chromatographed on a Sepharose 4 B column, electrotransferred on Immobilon P membranes, and assessed for MUC 1 and Lewis a and b structures using monoclonal antibodies. In 90% of examined patients, higher amounts of MUC 1 mucin were observed at the end of eradication treatment. Similar results for Lewis a and b structures were found. In the case of MUC 1 and Lewis b, the differences were statistically significant. Helicobacter pylori influences expression of the soluble form of MUC 1 mucin and Lewis a and b structures present in gastric juice. PMID- 18409004 TI - Bone density in the adolescent athlete. AB - Adolescence is a critical time for bone mass accrual, and increases in bone mass through puberty are dependent on rising levels of gonadal steroids, growth hormone and insulin like growth factor-1. Many high school girls are involved in athletic activities, and as many as 23.5% of adolescent athletes have been reported to develop amenorrhea. This review focuses on (1) factors that determine which athletes are likely to develop amenorrhea, such as a negative energy balance state, low levels of leptin and high levels of ghrelin, and (2) the impact of hypogonadism in athletes on bone metabolism. Beneficial effects of increased mechanical loading from athletic activity do not appear to protect against the deleterious effects of hypogonadism in adolescent athletes. PMID- 18409005 TI - Who are we missing? Area deprivation and survey participation. AB - This study investigates selective participation by area deprivation in two population-based, nationally-representative surveys of children aged 5-16 years in England. We analysed the English participants (N = 22,903) in the British Child and Adolescent Mental Health Surveys, conducted in 1999 and 2004. The surveys recruited parents, teachers and children, allowing us to calculate participation rates for all three types of informant, with particularly detailed analyses possible for parents. Area-deprivation was assigned using Indices of Multiple Deprivation, 2004. We found that parents, teachers and children from more deprived areas were all substantially less likely to participate, with participation 8-9% lower in the most deprived areas than in the least deprived areas. There was substantial heterogeneity in the effect of deprivation on different types of parental non-participation, with a strong association for overall non-contact but none for overall non-cooperation. Refusal to participate in response to opt-out letters showed the opposite gradient to all other forms of non-participation, with individuals from less deprived areas being more likely to opt out. These findings indicate that the deprivation level of area of residence predicts non-response in multiple types of informants, which may exacerbate bias when estimating the prevalence of health outcomes. Furthermore, the magnitude and the direction of this may depend on the recruitment method used. Differential response by area deprivation may therefore be worth measuring in any survey. PMID- 18409006 TI - The association between father's social class and adult obesity is not explained by educational attainment and an unhealthy lifestyle in adulthood. AB - OBJECTIVES: To investigate the effect of father's social class on central and general obesity in adulthood. To test the role of educational attainment and adult health behaviours as mediators in this association. METHODS: BMI, Waist-hip ratio, smoking status, alcohol consumption, leisure-time physical activity and dietary intake (semi-quantitative food-frequency questionnaire) were assessed at phase 5 (1997-1998) of the Whitehall II cohort study. We used retrospective data on educational attainment and father's social class. Our study sample was 4598 participants (3364 men, 1234 women) aged 44-69, with a valid obesity measure and information on father's social class and educational attainment. RESULTS: Father's social class was inversely associated with adult life central (Waist-hip ratio) and general (BMI) obesity in women but not in men. For example, mean BMI difference between the highest and lowest childhood social class was 2.04 kg/m(2) (95% confidence intervals: 0.90; 3.18). These associations remained robust to adjustment for adult life socioeconomic position. Adjusting for educational attainment resulted in a reduction of 10-15% in the difference in mean obesity measures between lowest and highest father's social class. In our fully adjusted model, adult life health behaviours did not provide further attenuation of the effect of father's social class on adult obesity. CONCLUSION: We provide evidence for an independent effect of father's social class on adult central and general obesity in women, which is not explained by educational attainment and an unhealthy lifestyle in adulthood. Policies aiming at reducing inequalities in obesity should tackle the problem of social inequality early in life. PMID- 18409007 TI - Alcohol consumption and risk of colorectal cancer: the Findrink study. AB - We investigated the association between alcohol consumption and colorectal cancer because previous studies have yielded conflicting results. As part of the Findrink study, data from the Kuopio Ischaemic Heart Disease (KIHD) Risk Factor Study were analysed. The KIHD study is a cohort of 2,682 men from Eastern Finland with no history of cancer at baseline. The men were grouped into five groups according to their weekly alcohol intake in grams. Association between alcohol and colorectal cancer was examined using Cox proportional hazard models. There were 59 cases of colorectal cancer during an average follow up of 16.7 years. Men within the highest quintile of alcohol consumption had a median weekly alcohol intake of 198.8 g. Age and examination year adjusted risk ratio of colorectal cancer among men within the highest quintile of alcohol consumption was 4.4 (95% CI: 1.6-11.9, P-value = 0.004). After adjusting for potential confounders, such as vegetable consumption, fibre intake, smoking, family history of cancer, socio economic status, leisure time physical activity, men with the highest amount of alcohol consumption still had a 3.5-fold (95% CI: 1.2-9.9, P-value = 0.021) increased risk of colorectal cancer. Exclusion of men diagnosed with colorectal cancer during the first 2 years of follow up from the analyses did not alter the risk increase. In conclusion, this study gives further evidence of a positive association between alcohol consumption and the risk of colorectal cancer. PMID- 18409008 TI - Genetic polymorphism of MTHFR G1793A in Chinese populations. AB - 5,10-methylenetetrahydrofolate reductase (MTHFR) is an important enzyme in folate metabolism. A novel polymorphic site in MTHFR (G1793A) could influence the homocysteine levels and was first described in 2002. Investigations revealed that this allele was associated with susceptibility to several cancers, but its distribution around the world was not adequate. To study the prevalence of the mutant frequency in Chinese populations, 923 healthy individuals from 13 Chinese populations distributing widely from north to south were collected. DNA samples were isolated from peripheral blood samples and genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), with the digestion of restriction endonuclease BsrBI. Of the 923 individuals, 82.1% were GG homozygous, 17.2% were GA heterozygous and 0.7% were AA homozygous. The frequency of the MTHFR 1793A allele in all tested individuals was 9.3%, which was slightly lower than indicated by HapMap (10%, Beiing Han, 45 samples). The frequencies of A allele were generally higher in southern China than that in northern China, and the frequencies had significant variance in 13 Chinese populations (X2 = 26.315, P = 0.010). Summarizing of the MTHFR G1793A allele polymorphism, including control groups in the case-control studies, we found only 20 normal peoples with AA homozygous (7 Chineses, 1 Caucasian, 2 Java Indonesias, 2 non-Hispanic whites, 6 Irish women, 2 Indians). The Java Indonesias and Ashkenzai Jevish had the highest (26.6%) and the lowest (1.3%) 1793A frequency, respectively. Together with our previous data, the MTHFR G1793A polymorphism was in linkage disequilibrium with both C677T and A1298C polymorphism sites in Chinese population, but not as strong as presented by HapMap. PMID- 18409009 TI - The M235T polymorphism of the AGT gene modifies the risk of coronary artery disease associated with the presence of hypercholesterolemia. AB - OBJECTIVE: Atherosclerosis is an inflammatory disease resulting from interactions between various genetic and non-genetic factors. Angiotensinogen gene (AGT) belongs to polymorphic candidate genes. Recent evidence show that many traditional risk factors of coronary artery disease (CAD) influence synthesis of AGT. This report focuses on the interactions between M235T polymorphism of AGT gene and traditional risk factors of CAD. MATERIAL AND METHODS: 255 subjects, including 158 patients with angiographically confirmed CAD and 97 blood donors without history of cardiovascular diseases were studied. M235T polymorphism of the AGT gene was genotyped using PCR-RFLP method. To determine the possible interactions of AGT genotypes and traditional risk factors of CAD the attributable proportion due to interaction (AP) and synergy models were used. RESULTS: The frequency of 235T allele carriers was significantly higher in patients than in controls (77.8 vs. 62.9, OR = 2.20, 95% CI; 1.10-4.40, P = 0.026, in multivariate logistic regression model). We found the existence of interaction between the 235T allele carrier-state and hypercholesterolemia (total cholesterol > or = 5 mmol/l) increasing the risk of CAD (SI = 3.39, 95% CI; 1.33 8.66, AP = 0.65, 95% CI; 0.39-0.91). The 235T allele also interacted with elevated LDL cholesterol levels (> or = 3 mmol/l) (AP = 0.49, 95% CI; 0.20-0.96), but not with the hypertension, overweight/ obesity and cigarette smoking. CONCLUSION: The 235T allele increases the risk of CAD associated with the presence of hypercholesterolemia. PMID- 18409010 TI - Re: Louwman WJ et al. On the rising trends of incidence and prognosis for breast cancer patients diagnosed 1975-2004: a long-term population-based study in southeastern Netherlands. Cancer Causes Control 19:97-106. PMID- 18409011 TI - Chromosome pairing in allotetraploid hybrids of Festuca pratensis x Lolium perenne revealed by genomic in situ hybridization (GISH). AB - Genomic in situ hybridization (GISH) was used to make a detailed study of chromosome pairing at metaphase I (MI) of meiosis in six F(1) hybrid plants of the allotetraploid Festuca pratensis x Lolium perenne (2n = 4x = 28; genomic constitution FpFpLpLp). The mean chromosome configurations for all hybrids analysed were 1.13 univalents + 11.51 bivalents + 0.32 trivalents + 0.72 quadrivalents, and the mean chiasma frequency was 21.96 per cell. GISH showed that pairing was predominantly intragenomic, with mean numbers of L. perenne (Lp/Lp) and F. pratensis (Fp/Fp) bivalents being virtually equal at 5.41 and 5.48 per cell, respectively. Intergenomic pairing between Lolium and Festuca chromosomes was observed in 33.3% of Lp/Fp bivalents (0.62 per cell), in 79.7% of trivalents - Lp/Lp/Fp and Lp/Fp/Fp (0.25 per cell), and in 98.4% of quadrivalents - Lp/Lp/Fp/Fp and Lp/Lp/Lp/Fp (0.71 per cell). About 4.0% of the total chromosome complement analysed remained as univalents, an average 0.68 Lp and 0.45 Fp univalents per cell. It is evident that in these hybrids there is opportunity for recombination to take place between the two component genomes, albeit at a low level, and this is discussed in the context of compromising the stability of Festulolium hybrid cultivars and accounting for the drift in the balance of the genomes over generations. We speculate that genotypic differences between hybrids could permit selection for pairing control, and that preferences for homologous versus homoeologous centromeres in their spindle attachments and movement to the poles at anaphase I could form the basis of a mechanism underlying genome drift. PMID- 18409012 TI - Evaluation of a fast wastewater odour characterisation procedure using a chemical sensor array. AB - Sewage treatment works are one of the major sources that cause atmospheric odour pollution. The increase in the number of complaints about odour nuisance is due to the increase in environmental concerns. Unfortunately, the legislation on odour nuisance from sewage treatment works is very limited. In order to determine suitable thresholds on which to base legal standards, reliable and efficient odour measurement methods need to be defined. A chemical sensor array was developed for the purpose of measuring wastewater odour. This paper describes the development of the chemical sensor system which is specifically tuned to odours of wastewater origin and which can give an electronic measure of the wastewater odours. Odour emissions from a wastewater treatment facility were detected by using a quartz crystal microbalance (QCM) sensor array. The array consists of nine sensor elements, which were coated with different materials. In this paper, the usage of these novel instruments in the water industry was shown. PMID- 18409013 TI - Water quality and heavy metal monitoring in water and sediment samples of the Kucukcekmece Lagoon, Turkey (2002-2003). AB - Physical and chemical parameters have been analyzed in water samples from a brackish water lagoon, Kucukcekmece, located on the western outskirts of Istanbul. Samples were collected every two months for a year from nine sampling stations. Of the parameters measured, temperature, pH, salinity, nitrate and phosphate showed changes when compared with the previously published data. The lagoon was found eutrophic as it was reported previously. Sulphate and COD levels were higher when compared with the standards established by the Turkish Water Pollution and Control Regulation. Additionally, concentrations of heavy metals (Cd, Co, Cr, Cu, Mn, Ni, Pb, Zn) in water and bottom sediments were measured and compared with the standards established by the Turkish Water Pollution and Control Regulation and with the previously published data. The results were analysed statistically with respect to location and any relationships between the concentration of the elements in corresponding water and sediment samples were examined. Principal Component Analysis of water samples allowed us to discriminate three areas affected mainly by heavy metal contamination, possibly due to industrial, commercial and/or urban activities. Generally, the concentrations of the heavy metals were higher at stations near the three estuaries, suggesting a direct influence of the three creeks on the pollution of the Kucukcekmece Lagoon. Although elevated levels of Cd were recorded in several water samples, it was not detected in sediment. On the other hand, a particularly high level of Cr pollution was recorded most of the water and sediment samples. PMID- 18409014 TI - Evaluation of potential habitat with an integrated analysis of a spatial conservation strategy for David's deer, Elaphurus davidians. AB - How to assess the potential habitat integrating landscape dynamics and population research, and how to reintroduce animals to potential habitats in environments highly human disturbed are still questions to be answered in conservation biology. According to behavioral research on Elaphurus davidians, we have developed a suitability index and a risk index to evaluate the potential habitats for the deer. With these indices, we conducted two transect assessments to evaluate the gradient change of the target region. Then, taking rivers as border lines, we tabulated the forest areas, high grassland area and total area and then compared the forest and high grassland area in each subregion. Furthermore, we computed the land use transfer matrix for the whole Yancheng coast during 1987 2000. We also computed human modified index (HMI) in six subregions. Lastly with a geographical information system support we obtained the spatial distribution of the indices and evaluation of the whole potential habitats from a neighborhood analysis. The transect assessment showed that the suitability of the coastal area was higher than that of the inland area for the deer, while the southern area was higher than the northern. Landscape metrics and HMI analysis showed that different landscape patterns and different anthropogenic disturbance existed within the region, and the increasing human disturbance was the key factor causing the pattern dynamics. The evaluation of potential habitats showed that there was an estimated carrying capacity of no more than 10,000 for David's deer reintroduction into the natural area. Also the reintroduction strategy was discussed. This integrated approach linked the population research and the landscape metrics, and the dataset with different scale; thus, it is an approach likely to be useful for the protection of other large animal in a landscape highly disturbed by humans. PMID- 18409015 TI - Emerging approaches for the syntheses of bicyclic imidazo[1,2-x]-heterocycles. AB - Imidazo-[1,2-x]heterocycles are versatile building blocks for use in both a 'drug hunters' quest to discover new leads and a chemical biologists search for effective molecular tools in 'cell perturbation' studies. At the front end of the drug discovery flow chart, the last 5-10 years have witnessed the discovery of new high-throughput methodologies which very quickly have enabled access to virtual libraries of these chemo-types in the realm of 10(7) derivatives. Interestingly, these often neglected cores in patent cooperation treaty (PCT) applications appear in several highly effective marketed drugs, completing the medicinal chemists search for clinical success. Such rigid chemo-types, all containing a bridgehead nitrogen atom, are thus poised for an ever increasing impact on the discovery and development of new molecular therapeutics. The following mini-review will briefly cover therapeutic utility, chemical methodologies and automation developed to enable preparation of arrays of these chemo-types in a high-throughput manner. Synthetic emphasis is placed on a 3 component-3-center isocyanide based multi-component reaction (IMCR), which spans solution, solid phase, flourous and microwave assisted organic synthesis. PMID- 18409016 TI - The relationship between depression-malnutrition and echocardiographic-blood pressure parameters in chronic hemodialysis patients. AB - AIMS: The aim is to research the relationship between the degree of depression malnutrition and inadequate volume control. METHODS: The mean age of the 52 patients was 55+/-14.6 years. Malnutrition score [subjective global assessment (SGA)] and depression score [Taiwanese Depression Questionnaire (TDQ)] of each chronic hemodialysis (CHD) patient were calculated. Then an echocardiographic assessment was carried out with the same device 1 day before the second HD session of the week. The 24 h blood pressure monitoring was performed 1 day before the second HD session of the week. RESULTS: TDQ scores (TDQS) were >or=19 in 41 and <19 in 11 patients. TDQS was found to be significantly high in women (p=0.01) who were older than 40 years (p=0.03) and who have low family income (p=0.03). TDQS was found to be significantly correlated with HD duration (p=0.034), vena cava inferior collapse index (p=0.02), malnutrition score (p=0.011), residual renal function (ml/day) (p=0.03), level of albumin (p=0.0009), and iron (p=0.015). A positive correlation was detected between TDQS and the ratio of mean nighttime blood pressure/mean daytime blood pressure (p=0.005, r=0.394). Depression score was found to be significantly different between normal geometry and left ventricular hypertrophy (eLVH), concentric remodeling and eLVH, and concentric LVH and eLVH. CONCLUSIONS: The results show that lesser degrees of nocturnal dip and eLVH are associated with increased degrees of depression, implying that hypervolemia is strongly associated with depression and might be a component of strong relationships involving malnutrition, inflammation, and atherosclerosis in CHD patients. PMID- 18409017 TI - A PCR based method to construct small interference RNA expression vectors. AB - Small interference RNAs (siRNA) have been shown to be useful in the field of gene therapy and gene function studies. As a siRNA expression vector, pSilencer employ RNA polymerase III promoters and could stably produce siRNA for weeks. But once one siRNA sequence was inserted into the pSilencer vector, the other siRNA sequence will hardly be reconstructed, because the site of siRNA production has been occupied and difficult to be changed, so it is not suitable for screen of effective siRNA sequence. To solve this problem, we constructed the subclone pSilcencer329, which generated from pSilencer3.1, then developed a PCR based method of constructing siRNA expression vectors, and generated pSilencerBCL2L2 recombinants efficiently. This method was proven to be effective, reliable, and less expensive, and thus will be of great help in regular gene silencing studies, and will be especially suitable for large scale gene function analysis. PMID- 18409018 TI - Focal lymphoid hyperplasia of the terminal ileum presenting mantle zone hyperplasia with clear cytoplasm. A report of three cases. AB - We report three unusual cases of focal lymphoid hyperplasia of the ileocecal valve. The gross specimens showed thickening of the ileocecal valve. Low power magnification showed a dense lymphoid infiltrate in the mucosa and submucosa. This condition was characterized by reactive lymphoid follicles with large reactive germinal centers surrounded by a pale cuff of mantle zone lymphocytes presenting a marginal zone distribution pattern. These cells had intermediate- to medium-sized round or slightly indented nuclei and a broad rim of clear cytoplasm. However, immunohistochemical study demonstrated that both the mantle zone lymphocytes and the pale cuff of the lymphoid cells were CD20+, sIgM+, sIgD+, CD5-, CD10-, CD23-, CD43-, Bcl-2+, Bcl-6-, CyclinD1-. The polytypic nature of these cells was demonstrated by immunohistochemistry and polymerase chain reaction. This unusual mantle cell hyperplasia with clear cytoplasm associated with focal lymphoid hyperplasia in middle-aged and elderly patients should be differentiated from the extranodal marginal zone B-cell lymphoma of mucosa associated lymphoid tissue type or mantle cell lymphoma showing a marginal zone distribution pattern. To avoid overdiagnosis and overtreatment, it is suggested that immunophenotypic and genotypic studies might be required, and careful attention should be paid to the morphologic examination. PMID- 18409019 TI - Radial scar-significant diagnostic challenge. AB - The prevalence of radial scar (RS) is 0.04% in asymptomatic women participating in population screening for breast cancer. It is important to differentiate RS from concomitant malignancies, which occur in 20-30% of patients, or from small stellate carcinomas which give similar radiomorphology. The aim of our study was to evaluate the effectivity of current breast diagnostic methods in distinguishing between real RS, concomitant malignancy and carcinomas imitating RS. Diagnosis of RS was set up in 61 cases by mammography. Forty-four patients underwent surgical excision: histology showed benign or malignant lesions in 28 and 16 cases, respectively. A series of negative results at follow-up proved the benign nature of the lesion in further 11 cases. Six patients were not available for follow-up. Results of mammography, physical examination, ultrasonography and cytology were evaluated and were compared in 39 benign and 16 malignant cases. Results of examinations were reported on the BI-RADS scale ranging from 1 to 5. The mean categorical scores of all diagnostic processes were around the level of borderline lesions: mammography: 3.49, ultrasonography: 3.06, cytology: 2.47 and physical examination: 1.67. The average age of the patients in the benign and malignant groups were the same: 58 years. The two groups did not differ significantly over either distribution of coded mammographical results (p = 0.2092), or the distribution of mammographical parenchyma density patterns (p = 0.4875). However, the malignant and benign groups differed significantly from each other over the distribution of coded ultrasonographic (p = 0.0176) and cytological (p < 0.0001) results. In conclusion, in the preoperative diagnosis of asymptomatic "black-stars", mammography detects the non-palpable lesions, and ultrasonography together with cytology proved better in the analysis, provided FNAB is US guided. Due to the complex diagnostic approach the nature of the "black stars" is known in the majority of cases prior to the surgical biopsy. PMID- 18409020 TI - Lapatinib: a sword with two edges. AB - Lapatinib is an oral dual tyrosine kinase inhibitor targeting EGFR1 and EGFR2 (HER2). Phase I trials have shown that lapatinib is well tolerated, with mild diarrhea and skin rush as common adverse effects, and low cardiotoxicity. Phase II and III trials provided evidences on clinical effectiveness in advanced or metastatic breast cancer and potential against brain metastases. Lapatinib is active in combination with trastuzumab and in trastuzumab-resistant patients, moreover it has synergistic action with capecitabine. Several clinical trials are in progress to explore the effectiveness of lapatinib in other combinations and against several tumor types. PMID- 18409021 TI - Seasonal variation of childhood acute lymphoblastic leukaemia is different between girls and boys. AB - The aim of this study was to investigate seasonal trends in the incidence of acute lymphoblastic leukaemia (ALL) around the times of birth and diagnosis in children aged 0-4 years and also to examine gender specific effects. Children born in South Hungary during 1981-1997 were analysed. Registrations of first malignancies for children, diagnosed under age 5 years before the end of 2002 were obtained from the Hungarian Paediatric Oncology Group providing a representative sample of Hungarian children over a 17 year period of time. Data were available on the corresponding numbers of births for each month of the study period were obtained. Statistical analyses were performed using logistic regression with harmonic components. The study analysed 121 cases of children, aged under 5 years, who were diagnosed with ALL. We found no seasonal effect related to date of diagnosis. However, there was seasonal variability for ALL related to date of birth. Maximal rates were seen in children born in February and August in the simple harmonic regression model for all children diagnosed with ALL. Analysis by gender found evidence of seasonality related to month of birth with peaks in February and August in boys, but different seasonal effects were seen for girls (peak in November, nadir in May). Our study provides some evidence that male specific immune responses to infections around the time of birth could explain the male predominance in the incidence of ALL. PMID- 18409022 TI - Primary adamantinoma of the rib. Unusual presentation for a bone neoplasm of uncertain origin. AB - Adamantinomas are rare, low-grade malignant intra-osseous tumors composed of epithelial and mesenchymal elements, which show a marked predilection for the tibia and fibula of young adult male patients. Although cases of adamantinoma located to the axial skeleton have been reported either as recurrent or metastatic disease, only two cases of primary adamantinoma located to the thoracic wall have been previously described. In this study we present the clinical, radiological and histopathological features of a 24-year-old male with a slow growing, solid-cystic, painful mass, located to the right 11th rib, which was morphological and immunohistochemically diagnosed as a primary classic adamantinoma. Radiological studies showed a multiloculated lesion with a solid component. The patient underwent a whole surgical resection of the lesion. Histologically, multiple foci of epithelial cells with basaloid and squamous components were found intermixed within a fibrous stromal tissue. Immunohistochemical analysis demonstrated expression of cytokeratins, EMA, vimentin and other epithelial markers. Primary affection of the rib is an unusual feature of classic adamantinomas. PMID- 18409023 TI - Granule cell survival is deficient in PAC1-/- mutant cerebellum. AB - PACAP exerts neuroprotective effects during development, especially in the cerebellum where PAC1 receptor and ligand are both expressed. However, while previous studies using PACAP injections in postnatal animals defined trophic effects of exogenous peptide, the role of endogenous PACAP remains unexplored. Here, we used PAC1(-/-) mice to investigate the role of PACAP receptor signaling in postnatal day 7 cerebellum. There was no difference in DNA synthesis in the cerebellar EGL of PAC1(-/-) compared to wild type animals, assessed using thymidine incorporation and BrdU immunohistochemistry. In contrast, we found that a significant proportion of newly generated neurons were eliminated before they successfully differentiated in the granule cell layer. In aggregate, these results suggest that endogenous PACAP plays an important role in cell survival during cerebellar development, through the activation of the PAC1 receptor. PMID- 18409024 TI - Exploration of the endogenous agonist mechanism for activation of secretin and VPAC1 receptors using synthetic glycosylated peptides. AB - Current understanding of the molecular basis of activation of class II G protein coupled receptors remains limited, despite recent solution of NMR and crystal structures of amino-terminal domains of several family members. One mechanism proposed for the activation of these receptors involves an agonist-stimulated change in conformation of the receptor amino terminus. This results in the exposure of a "hidden endogenous agonist" (WDN sequence in secretin and VPAC1 receptors) within the receptor amino terminus that interacts with the receptor core, thereby changing its conformation and exposing its G protein-binding region. The Asn in this WDN sequence is known to be glycosylated in both secretin and VPAC1 receptors, raising concern about whether this posttranslational modification might interfere with the proposed mechanism. Therefore, we prepared glycosylated forms of cyclic WDN and the longer cyclic peptide, LWDNM, and tested them for agonist activity at secretin and VPAC1 receptor-bearing cell lines. Both glycosylated peptides stimulated full cAMP responses in the cell lines. Clearly, glycosylation did not interfere with this mechanism and may actually facilitate the correct orientation of the pharmacophore of the endogenous agonist ligand. These data provide further evidence for this proposed mechanism for the activation of this family of receptors. PMID- 18409025 TI - A practical diagnostic approach to solid/trabecular nodules in the thyroid. AB - Nodules having a non-follicular-non-papillary structure may be frequently encountered in the thyroid, the solid/trabecular arrangement being the most common feature. When solid/trabecular pattern is predominant within a thyroid nodule, the wide range of differential diagnoses represents one of the major diagnostic problems in the routine thyroid practice, including--among others- relatively frequent entities such as trabecular adenoma or solid variant of papillary carcinoma and rare tumors such as hyalinizing trabecular tumor or paraganglioma. Morphology alone may not be sufficient to proper-type solid/trabecular tumors in the thyroid and, therefore, a panel of immunohistochemical markers is recommended, including thyroglobulin/calcitonin as the first step, followed by specific markers according to the morphological and clinical setting. A schematic algorithm is presented, which takes into consideration the diagnostic hallmarks of the most relevant entities that enter in the differential diagnosis in solid/trabecular nodules within the thyroid. PMID- 18409026 TI - Characterization and expression of an actin-depolymerizing factor from Eimeria tenella. AB - Actin-depolymerizing factor (ADF) plays an important role in remodeling the actin cytoskeleton which contributes much to the invasion of host cells by the apicomplexan parasite. The gene encoding for Eimeria tenella ADF with one intron was cloned and identified by the E. tenella genome raw sequence data ( http://www.sanger.ac.uk/projects/E.tenella/ ). The deduced polypeptide sequence was only composed of 118 amino acids (13.14 kDa) without signal peptide and nuclear localization sequence. The amino acid sequence was most similar to the ADF of Toxoplasma gondii, 69.1%. Compared the putative three-dimensional structures between E. tenella and yeast, the actin filament binding sites on the segment from the alpha4-helices to the C-terminal were mostly missed in E. tenella. Real-time RT-PCR and dot blot both revealed that ADF expression was relatively stronger in the sporozoites and merozoites than in sporulated and unsporulated oocysts in both mRNA and protein levels. Northern blot analysis suggested that there was only one form of ADF transcripts in all different life stages of E. tenella. Actin-binding experiment showed that the recombinant fusion ADF protein could bind with actin, which indicated that ADF probably plays an important role in the invasion host of E. tenella. PMID- 18409027 TI - Site specific cytosine methylation in rice nonautonomous transposable element nDart. AB - The mobile nonautonomous element nDart, which is active in intact rice plants, exhibits locus specific transposition. Due to the high homogeneity of nDart elements, the locus specificity of nDart transposition might be controlled by factors other than genetic differences. In this study, we elucidated the regulation of the locus specificity of nDart transposition. The difference of transpositional activities in 10 nDart elements among rice varieties exhibiting nDart transposition was clearly correlated with the methylation state of nDart elements. Both hyper- and hypo-methylated nDart elements were inactive, while site specific methylation in both subterminal regions was identified in active nDart loci. The specific methylation sites contain the pentamer motif GCC/ACG. The repeated motifs in the subterminal region of nDart elements may contribute to the stable secondary structure of nDart elements with low free energy. Our results suggested that site specific cytosine methylation may loosen the stable secondary structure of the nDart element to allow it to bind TPase, which then perform the excision of nDart elements from genomic loci. PMID- 18409028 TI - Purification, cloning, functional expression and characterization of perakine reductase: the first example from the AKR enzyme family, extending the alkaloidal network of the plant Rauvolfia. AB - Perakine reductase (PR) catalyzes an NADPH-dependent step in a side-branch of the 10-step biosynthetic pathway of the alkaloid ajmaline. The enzyme was cloned by a "reverse-genetic" approach from cell suspension cultures of the plant Rauvolfia serpentina (Apocynaceae) and functionally expressed in Escherichia coli as the N terminal His(6)-tagged protein. PR displays a broad substrate acceptance, converting 16 out of 28 tested compounds with reducible carbonyl function which belong to three substrate groups: benzaldehyde, cinnamic aldehyde derivatives and monoterpenoid indole alkaloids. The enzyme has an extraordinary selectivity in the group of alkaloids. Sequence alignments define PR as a new member of the aldo keto reductase (AKR) super family, exhibiting the conserved catalytic tetrad Asp52, Tyr57, Lys84, His126. Site-directed mutagenesis of each of these functional residues to an alanine residue results in >97.8% loss of enzyme activity, in compounds of each substrate group. PR represents the first example of the large AKR-family which is involved in the biosynthesis of plant monoterpenoid indole alkaloids. In addition to a new esterase, PR significantly extends the Rauvolfia alkaloid network to the novel group of peraksine alkaloids. PMID- 18409029 TI - Nucleic acids bind to nanoparticulate iron (II) monosulphide in aqueous solutions. AB - In the hydrothermal FeS-world origin of life scenarios nucleic acids are suggested to bind to iron (II) monosulphide precipitated from the reaction between hydrothermal sulphidic vent solutions and iron-bearing oceanic water. In lower temperature systems, the first precipitate from this process is nanoparticulate, metastable FeSm with a mackinawite structure. Although the interactions between bulk crystalline iron sulphide minerals and nucleic acids have been reported, their reaction with nanoparticulate FeSm has not previously been investigated. We investigated the binding of different nucleic acids, and their constituents, to freshly precipitated, nanoparticulate FeSm. The degree to which the organic molecules interacted with FeSm is chromosomal DNA > RNA > oligomeric DNA > deoxadenosine monophosphate approximately deoxyadenosine approximately adenine. Although we found that FeSm does not fluoresce within the visible spectrum and there is no quantum confinement effect seen in the absorption, the mechanism of linkage of the FeSm to these biomolecules appears to be primarily electrostatic and similar to that found for the attachment of ZnS quantum dots. The results of a preliminary study of similar reactions with nanoparticulate CuS further supported the suggestion that the interaction mechanism was generic for nanoparticulate transition metal sulphides. In terms of the FeS-world hypothesis, the results of this study further support the idea that sulphide minerals precipitated at hydrothermal vents interact with biomolecules and could have assisted in the formation and polymerisation of nucleic acids. PMID- 18409030 TI - Involvement of Ad4BP/SF-1, DAX-1, and COUP-TFII transcription factor on steroid production and luteinization in ovarian theca cells. AB - To examine the essential mechanisms of steroid production in ovarian theca cells, we analyzed the expression of genes associated with steroid production using simple culture system with serum medium. In addition, we examined the involvement of DAX-1, COUP-TFII, and Ad4BP/SF-1 transcription factors on the steroid production in theca cells. Theca cells begin to display an elongated or fibroblastic aspect within 24 h of culture. Over the next 48 h, they metamorphosed from the fibroblastic to the epitheloid phenotype. The number of theca cells increased during culture period. Androstenedione and progesterone production per cell decreased at 48-96 h compared with 0-48 h of culture. Steroidogenic acute regulatory protein (StAR) and CYP 17 genes expression decreased at 48 h compared with 0 h of culture, and afterward maintained a low level. In contrast, expression of 3beta-HSD and P450scc mRNAs increased at 48 h compared with 0 h of culture. Protein expression of Ab4BP/SF-1 maintained a constant level during culture. COUP-TFII protein expression showed a peak level at 24 h of culture period. DAX-1 protein expression began to increase at 48 h of culture. Our data suggested that the inhibition in CYP 17 and StAR genes by DAX-1 transcription factor may be associated with the decrease in androstenedione and progesterone production by theca cells during in vitro culture. Such an essential pathway for steroid production might indicate the importance of theca cell function in bovine ovary. PMID- 18409031 TI - Functional genomics of hTERT gene in leukemic myelopoiesis. AB - Keeping in view the fact that a single acquired genetic abnormality "Bcr-Abl chimeric gene" accompanied by elevated telomerase activity has been widely recognized to be responsible for the leukemic myelopoiesis observed in chronic myeloid leukemia (CML), the present study was addressed to understand as to how selective and specific knock-down of human telomerase reverse transcriptase (hTERT) gene within mononuclear cells derived from untreated CML subjects could influence the apoptotic, genotypic (such as Bcr-Abl; C-myc; Bcl-2; IL-6; GMCSF; IL-3; and acetylated H(3) and H(4)), and phenotypic (such as CD34 and CD89) characteristics of these cells. Based upon these results, we propose that hTERT gene-based drug design may be useful in the treatment of leukemic myelopoiesis. PMID- 18409032 TI - EPR studies on the superoxide-scavenging capacity of the nutraceutical resveratrol. AB - Resveratrol (3,4',5-trihydroxystilbene), a polyphenolic compound found in mulberries, grapes, and red wine, has received considerable attention because of its apparent protective effects against various degenerative diseases due to its potential antioxidant activities. However, direct evidence for the superoxide scavenging capacity of resveratrol is lacking in literature. In this study, electron paramagnetic resonance spectroscopy in combination with 5 (diethoxyphosphoryl)-5-methylpyrroline-N-oxide (DEPMPO)-spin trapping technique was utilized to determine the ability of resveratrol in scavenging superoxide anions generated from both potassium superoxide and the xanthine oxidase/xanthine system. We have demonstrated here for the first time that the presence of resveratrol resulted in decreased formation of DEPMPO-superoxide adduct (DEPMPO OOH) in both the potassium superoxide and xanthine oxidase/xanthine systems, indicating that resveratrol could directly scavenge superoxide anions. The inhibition of DEPMPO-OOH in the xanthine oxidase/xanthine system, however, was found to be much potent as compared to that observed in potassium superoxide system. It was further shown that resveratrol could also directly inhibit xanthine oxidase activity as assessed by oxygen consumption and formation of uric acid. Taken together, the dual role of resveratrol in directly scavenging superoxide and inhibiting its generation via xanthine oxidase reported in this study may explain, at least in part, the protective role of this compound against oxidative injury in various disease processes. PMID- 18409033 TI - Hypertrophy of intima media of the carotid artery due to L-dopa therapy in Parkinson's disease. AB - INTRODUCTION: Recent studies have shown that L-dopa therapy used in patients with Parkinson's disease might cause thickening of the carotid artery intima media. It is known that intima-media thickening is a sign of systemic atherosclerosis. The purpose of this study was to clarify the correlation between L-dopa medication and carotid artery intima-media thickness (IMT) in patients with Parkinson's disease. METHODS: Twenty-five patients with Parkinson's disease who used L-dopa (LD+) and 17 patients who did not use L-dopa (LD-) treatment were included in this study. Colour Doppler with B-mode carotid ultrasonography was performed. The Hoehn-Yahr scale was used for staging of Parkinson's disease in an 'on' state. The Student t test was used for statistical analysis. RESULTS: The IMT of the left internal carotid and external carotid arteries were found to be thicker in patients treated with L-dopa. The mean IMT of the left internal carotid artery was 0.937+/-0.391 mm in the LD+ group and 0.677+/-0.138 mm in the LD-group (P=0.013). The mean IMT of the left external carotid artery was 0.956+/-0.531 mm in the LD+ group and 0.646+/-0.18 mm in the LD-group (P=0.037). CONCLUSION: The IMT of the carotid artery was found to be thicker in patients who were treated with L-dopa. In conclusion, patients treated with L-dopa appear to be at a higher risk of thickening of the intima media of the carotid artery. PMID- 18409035 TI - Total hip arthroplasty in dwarfism. A case report. AB - In dwarfism hip arthritis, usually secondary to hip dysplasia, is a common finding at an early age. In these patients a joint replacement is a demanding procedure due to the peculiar joint deformity and the small size of the bones. We present a case of a bilateral hip replacement in a dwarf patient. In order to reduce intraoperative complications and improve the joint kinematics a thorough preoperative planning was performed by a CT based computerised system. On the basis of the planning we chose a conical shaped stem that enable as to restore limb length and offset with a low risk of femoral fracture. In conclusion, we consider total hip replacement in dwarfism a safe and effective procedure if an accurate preoperative planning is performed. PMID- 18409034 TI - Degeneration of Paget's disease into sarcoma: clinical and therapeutic influencing factors. AB - We analysed a series of 119 patients presenting with Paget's bone disease treated at the Istituto Ortopedico Rizzoli (Bologna Italy) over 35 years (from 1970 to 2006). Among these, we found 18 cases that had degenerated into sarcoma. The first goal of this study was to underline the potential factors of Paget's disease transformation into sarcoma. In detail we considered: age, gender, duration of disease, clinical expression, levels of total alkaline phosphatase and type of therapy. The results highlighted an interesting correlation between therapy and degeneration into sarcoma; more specifically, malignant transformation occurred both in patients who had not received any therapy and those who had received regular calcitonin treatment; otherwise, no sarcoma degeneration occurred in the patients treated with bisphosphonates. These data seem to indicate a preventing effect of bisphosphonates towards the degeneration of Paget's disease into sarcoma. PMID- 18409036 TI - Anterolateral soft tissue sparing approach to the hip. Surgical technique. AB - The long-term successful outcomes of total hip replacement led to an extension of the surgical indications to young and active patients. To achieve a prompt functional recovery, varying surgical techniques have been proposed. In this study we present our anterolateral soft tissue sparing approach characterised by a lesser dissection of the muscles, a capsular repair and an accurate soft tissue tensioning and reconstruction. The target of this procedure is a full and quick restoration of hip function, reducing the rate of early complications such as hip dislocation. PMID- 18409037 TI - Biochemical pharmacology of biflavonoids: implications for anti-inflammatory action. AB - Biflavonoids belong to a subclass of the plant flavonoid family. Distribution of biflavonoids in the plant kingdom is limited to several species. Previously, some pharmacological activities of biflavonoids were described such as inhibition of histamine release from mast cells and inhibition of lymphocyte proliferation, suggesting the anti-inflammatory/antiallergic potential of the biflavonoids. Furthermore, several natural biflavonoids including ochnaflavone and ginkgetin inhibit phospholipase A2. Most importantly, certain biflavonoids exhibit anti inflammatory activity through the regulation of proinflammatory gene expression in vitro and in vivo. Recently, several synthetic approaches yielded new biflavonoid molecules with anti-inflammatory potential. These molecules also exhibit phospholipase A2 and cyclooxygenase-2 inhibitory activity. Although the bioavailability needs be improved, certain biflavonoids may have potential as new anti-inflammatory agents. This is the first review of biflavonoid pharmacology to date. PMID- 18409038 TI - Antioxidative compounds from Quercus salicina Blume stem. AB - The chromatographic separation of MeOH extract from the Quercus salicina Blume Stem led to the isolation of five phenolic compounds. Using spectroscopic methods, the structures of these compounds were determined as D-threo guaiacylglycerol 8-O-beta-D-(6'-O-galloyl)glucopyranoside (1), 9-methoxy-D-threo guaiacylglycerol 8-O-beta-D-(6'-O-galloyl)glucopyranoside (2), 6''-O-galloyl salidroside (3), methyl gallate (4), quercetin (5). We measured radical scavenging activity with the DPPH method and the anti-lipid peroxidative efficacy on human LDL with TBARS assay, with the result that all these compounds exhibited the antioxidative activity. PMID- 18409039 TI - Synthesis and antimicrobial activity of certain new 1,2,4-triazolo[1,5 a]pyrimidine derivatives. AB - Certain new derivatives of 1,2,4-triazolo[1,5-a]pyrimidines were synthesized through the reaction of 1,2,4-triazolo[1,5-a]pyrimidine-7-ol with ethyl bromoacetate to afford the ethyl acetate ester, which upon hydrazinolysis gives the corresponding hydrazide. The hydrazide is the key intermediate which was used for the synthesis of the target compounds. The structures of the new compounds were assigned by spectral and elemental methods of analyses. The synthesized compounds were tested for their in vitro antibacterial and antifungal activities. Most of the tested compounds showed comparable results with those of ampicillin and fluconazole reference drugs. PMID- 18409041 TI - Synthesis of DAAS derivatives and their enhancement of HL-60 leukemia cell differentiation. AB - DAAS is the diacetoxy acetal derivative of a-santonin and induces HL-60 cell differentiation into granulocytes. In this report, we investigated the structure activity relationship (SAR) of DAAS derivatives in the differentiation of human HL-60 leukemia cells. Although its derivatives themselves had less effect on HL 60 cell differentiation than DAAS, the monoacetyl derivative, 2, mainly induced HL-60 cell differentiation. Moreover, compound 2 synergistically enhanced all trans retinoic acid (ATRA)-induced HL-60 cell differentiation when combined with 50 nM ATRA, a well-known differentiation inducer. This enhancing effect is similar to that of DAAS in ATRA-induced differentiation. PMID- 18409040 TI - Melanogenesis inhibitory compounds from Saussureae Radix. AB - Ten compounds were isolated from the EtOAc soluble part of the MeOH extract of Saussureae Radix, with their effects on melanin production also evaluated in B-16 mouse melanoma cell lines stimulated with 3-isobutyl-1-methylxanthine (IBMX), an elevator of cellular cAMP. The compounds were identified as aplotaxene (1), 1 beta-hydroxy arbusculin A (2), costunolide (3), dehydrocostuslactone (4), 11 beta,13-dihydrocostunolide (5), reynosin (6), heptadec-(9Z)-enoic acid (7), beta sitosterol (8), linoleic acid methyl ester (9) and betulinic acid methyl ester (10). Compounds 2, 9 and 10 were identified from Saussureae Radix for the first time. Furthermore, compounds 2, 3 and 6 showed potent inhibitory effects on the IBMX-induced melanogenesis, in dose-dependent manners, with IC50 values of 11, 3 and 2.5 microg/mL, respectively. As a positive control, arbutin exhibited an IC50 value of 29 microg/mL. PMID- 18409042 TI - Practical demethylation of aryl methyl ethers using an odorless thiol reagent. AB - A highly practical method for demethylation of aryl methyl ethers employing a long-chain thiol has been developed. Under the conditions described herein, clean and fast conversions to the desired phenolic compounds have been achieved with a broad range of substrates. Unlike other thiolate-mediated methods, this newly developed protocol features in-situ generation of sodium alkylthiolate using NaOH, and is almost free from foul smells and potentially harmful gases. It therefore provides an attractive option for the demethylation of aryl methyl ethers. PMID- 18409043 TI - Lysophosphatidylserine induces calcium signaling through Ki16425/VPC32183 sensitive GPCR in bone marrow-derived mast cells and in C6 glioma and colon cancer cells. AB - Lysophosphatidylserine (LPS) can be generated following phosphatidylserine specific phospholipase A2 activation. The effects of LPS on cellular activities and the identities of its target molecules, however, have not been fully elucidated. In this study, we observed that LPS stimulated intracellular calcium increased in mouse bone marrow-derived mast cells (BMMC), and rat C6 glioma and human HCT116 colon cancer cells and compared the LPS-induced Ca2+ increases with the response by lysophosphatidic acid (LPA), a structurally related bioactive lysolipid. In order to test involvement of signaling molecules in the LPS-induced Ca2+ signaling, we used pertussis toxin (PTX), U73122, and 2-APB, which are specific inhibitors for G proteins, phospholipase C (PLC), and IP3 receptors, respectively. The increases due to LPS and LPA were inhibited by PTX, U-73122 and 2-APB, suggesting that both lipids stimulate calcium signaling via G proteins (Gi/o types), PLC activation, and subsequent IP3 production, although the sensitivity to pharmacological inhibitors varied from complete inhibition to partial inhibition depending on cell type and lysolipid. Furthermore, we observed that Ki16425 completely inhibited an LPS-induced Ca2+ response in three cell types, but that the effect of VPC32183 varied from complete inhibition in BMMC and C6 glioma cells to partial inhibition in HCT116 cells. Therefore, we conclude that LPS increases [Ca2+]i through Ki16425/VPC32183-sensitive G protein-coupled receptors (GPCR), G protein, PLC, and IP3 in mouse BMMC, rat C6, and human HCT116 cells. PMID- 18409044 TI - Chemical constituents of Melandrium firmum Rohrbach and their anti-inflammatory activity. AB - In our ongoing search for anti-inflammatory agents originating from Korean medicinal plants, we found that the hexane and BuOH fractions of the MeOH extract from the whole plants of Melandrium firmum Rohrbach inhibited 5-lipoxygenase (5 LOX) activity. By activity-guided fractionation, eleven compounds, alpha spinaterol (1), ursolic acid (2), ergosterol peroxide (3), alpha-spinaterol glucoside (4), 2-methoxy-9-beta-D-ribofuranosyl purine (5), aristeromycin (6), ecdysteron (7), polypodoaurein (8), (-)-bornesitol (9), mannitol (10) and cytisoside (11) were isolated from the hexane and BuOH fractions using column chromatography. Compounds 2, 5, 6, 8, 9, 10 and 11 were isolated for the first time from this plant. Compounds 1, 3, 4 and 7 inhibited 5-LOX activity with IC50 values of 21.04 microM, 42.30 microM, 32.82 microM, and 17.18 microM, respectively. PMID- 18409045 TI - A new triterpenoid from Panax ginseng exhibits cytotoxicity through p53 and the caspase signaling pathway in the HepG2 cell line. AB - A new triterpenoid, 20(R),22(xi),24(S)-dammar-25(26)-ene-3beta,6 alpha,12 beta,20,22,24-hexanol (1), and three known triterpenoids, beta-D glucopyranoside,(3beta,12 beta)-12,20-dihydroxydammar-24-en-3-yl,6-acetate (2), 20(R)-ginsenoside Rg3 (3), and 20(R)-ginsenoside Rh2 (4), were isolated from the leaves of Panax ginseng. Their structures were determined by chemical analysis and spectral methods (IR, 1D and 2D NMR, HR-ESI-MS). Compounds 1-4 were exhibited various degrees of cytotoxicity in the human hepatoma cell line, HepG2. Compound 1 had the highest cytotoxic potency, with an IC50 value of 20.1 microM, by stimulating p53-mediated cell cycle arrest at the G1 to S phase transition, leading to apoptosis via activation of the caspase signaling pathway. PMID- 18409046 TI - NMR-based metabolomics approach for the differentiation of ginseng (Panax ginseng) roots from different origins. AB - Agro-herbal materials vary in prices and qualities depending on the origin and age and the differentiation is both scientific and public health issue. Here, we describe a metabolomics approach used to discriminate ginseng roots from different sources. Six different types of ginseng roots from China and Korea were analyzed by NMR-based metabolomics. Chinese Dangsam showed prominent differences and was easily differentiated. The difference was mainly due to the large signals in the sugar region. We further analyzed the metabolomics results in subgroups. Jeonra (Korean), Choongcheong (Korean), and Chinese ginseng in subgroup 1 could be easily differentiated by the first two principal components. The loading plot for PC1 showed that the Jeonra and Chinese ginseng roots were mainly separated by sugar signals and methyl signals but that they were reverse-correlated. A diffusion-ordered spectroscopy (DOSY) analysis showed that the methyl signals are from high molecular weight compounds and that the sugar signals are either from oligosaccharides or ginsenosides. In subgroup 2, composed of Korean Choongcheong ginseng at different ages, we were able to see age-dependent transitions in the score plot. We believe our approach can be applied to detecting the adulteration of ginseng root powders and other herbal products from different origins. PMID- 18409047 TI - Hydroquinone, a major component in cigarette smoke, reduces IFN-gamma production in antigen-primed lymphocytes. AB - Exposure to cigarette smoke is known to suppress immune responses and to increase the incidence and severity of respiratory infections. In this study, we determined the effect of hydroquinone (HQ), which is found at high concentrations in cigarette smoke, on interferon-gamma (IFN-gamma) production by lymphocytes. HQ significantly inhibited IFN-gamma secretion by keyhole limpet hemocyanin-primed lymphocytes in a dose-dependent manner. In addition, HQ inhibited IFN-gamma secretion in effector CD4+ T cells and Th1-differentiated CD4+ T cells. The mRNA expression of IFN-gamma and the IFN-gamma gene promoter activity were inhibited by HQ. These results suggest that the inhibitory effect of HQ on IFN-gamma secretion may occur at the transcriptional level. Furthermore, the effects of HQ on transcription factors were investigated. HQ inhibited the transcriptional activity of activator protein-1 and nuclear factor-kappa B, which are known to be involved in IFN-gamma transcriptional activation. These findings provide evidence that HQ might suppress immune responses by reducing the production of IFN-gamma and may explain the susceptibility to microbial infections caused by cigarette smoking. PMID- 18409048 TI - Inhibitory effect of citral on NO production by suppression of iNOS expression and NF-kappa B activation in RAW264.7 cells. AB - Citral is a major compound of lemongrass (Cymbopogon citratus L.) that has many pharmacological activities such as anti-fungal and anti-bacterial effects. In this study, we investigated the anti-inflammatory effect of citral and defined its mechanism of action in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Citral (3-12 microg/mL) significantly inhibited LPS-induced nitric oxide (NO) production in a concentration-dependent manner (IC50: 6.5 microg/mL). Furthermore, it was found that citral effectively inhibited the transcriptional activity and expression of iNOS, and potently suppressed the DNA binding activity and nuclear translocation of NF-kappa B as well as I kappa B phosphorylation in a concentration dependent manner. These results suggest that citral is anti inflammatory, and its effects may be due to the inhibition of NO production through the suppression of NF-kappa B activation. PMID- 18409049 TI - Differential regulation of ErbB2 expression by cAMP-dependent protein kinase in tamoxifen-resistant breast cancer cells. AB - Acquired resistance to tamoxifen (TAM) is a serious therapeutic problem in breast cancer patients, and Her-2/ErbB2 expression is associated with decreased sensitivity to TAM. We previously reported that cAMP-dependent protein kinase (PKA)-mediated activator protein-2 (AP-2) activation was responsible for the expression of Her-2/ErbB2 in p53-inactivated mammary epithelial cells (Yang et al., 2006). In the present study, we tested the hypothesis that PKA plays a role in the expression of ErbB2 in tamoxifen-resistant breast cancer cells. Treatment with H-89, a specific PKA inhibitor, suppressed 4-hydroxytamoxifen-induced ErbB2 expression in control MCF-7 cells. In contrast, PKA inhibition by H-89 or cAMP dependent protein kinase inhibitor l gamma overexpression increased the expression levels of ErbB2 in TAM-resistant MCF-7 (TAMR-MCF-7) cells. Transcriptional regulation of the erbB2 gene depends on two transcription factors, AP-2 and polyomavirus enhancer activator3 (PEA3). H-89 decreased nuclear or total levels of PEA3 in TAMR-MCF-7 cells. Chromatin immunoprecipitation assay results revealed that H-89 treatment reduced PEA3 binding to the proximal Ets binding site of the erbB2 gene promoter. Reporter gene analyses using human erbB2 gene promoter supported the critical role of PEA3 in the overexpression of ErbB2 in TAMR-MCF-7 cells treated with H-89. This deregulated PKA signaling cascades required for the ErbB2 expression may be important for the differential response of TAM-resistant breast cancer cells to EGF/ErbB2 stimuli. PMID- 18409050 TI - Inhibitory effects of crocetin on high glucose-induced apoptosis in cultured human umbilical vein endothelial cells and its mechanism. AB - Dysfunction of endothelial cell is considered as a major cause of vascular complications in diabetes. Crocetin has been shown to have strong antioxidant activities. In present study, we tested whether crocetin inhibited high glucose induced apoptosis in cultured human umbilical vein endothelial cells (HUVECs) and to explore its possible mechanism. Exposure to high glucose (33 mM) for 72h induced a pronounced increase in apoptosis compared with normal glucose (5.5 mM), as evaluated by cell chromatin staining with Hoechst 33,258 and cell death detection ELISA. High glucose attenuated activation of Akt and endothelial nitric oxide synthase (eNOS). Crocetin (0.1 microM, 1.0 microM) prevented high glucose induced apoptosis, which correlates with the increase of activation of p-Akt, following the up-regulation of eNOS and NO production. Pretreatment with phosphatidylinositol 3' kinase (Pl3K) inhibitor LY294002 or eNOS inhibitor NG nitro-arginine methyl ester (LN or L-NAME) inhibited crocetin effect on p-Akt or eNOS, respectively. For the first time, results of our study suggest that crocetin inhibits high glucose-induced apoptosis, at least partly, via Pl3K/Akt/eNOS pathway in HUVECs and crocetin may exert a beneficial effect in preventing diabetes-associated cardiovascular complications. PMID- 18409051 TI - Intracerebroventricular ginsenosides are antinociceptive in proinflammatory cytokine-induced pain behaviors of mice. AB - Several ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, Rg1 and Rg3) are neuroprotective and antinociceptive agents. In this study, we assessed the effects of these ginsenosides following intracerebroventricular (i.c.v.) administration on the nociceptive behaviors induced by intrathecal injection of pro-inflammatory cytokines (tumor necrosis factor-a (TNF-alpha), interleukin-1 beta (IL-1 beta), and interferon-gamma (IFN-gamma)). The ginsenosides, Rb1, Rb2, Rc, Rd, Re, Rf and Rg1, significantly attenuated the nociceptive behavior induced by TNF-alpha, IL-1 beta, and IFN-gamma injection, but ginsenoside-Rg3 did not. These results suggest that several ginsenosides may regulate the nociceptive processing induced by pro inflammatory cytokines. PMID- 18409053 TI - Molecular analysis of constitutive mutations in ermB and ermA selected in vitro from inducibly MLSB-resistant enterococci. AB - Frequencies of spontaneous mutation from inducible resistance to constitutive resistance were determined for the four clinical isolates of erythromycin resistant enterococci, including one isolate with ermB gene and three clinical isolates with ermA gene. The rate of ermB mutation was higher than that of ermA mutation by more than 10 fold. Sequence analysis of the regulatory regions of erm genes revealed that mutation type of ermB was just point mutation, by contraries the mutation type of ermA was either deletion or tandem duplication. These results showed distinct characteristics in mutation patterns of ermB and ermA. PMID- 18409052 TI - Auranofin, an immunosuppressive drug, inhibits MHC class I and MHC class II pathways of antigen presentation in dendritic cells. AB - Auranofin (AF), an orally administered, gold-based, anti-arthritic agent, has emerged as a clinically useful therapeutic drug for the treatment of rheumatoid arthritis. In the present study, we examined the effects of AF on major histocompatibility complex (MHC)-restricted antigen presentation in dendritic cells (DCs), which are the most important accessory cells for the induction of T cell responses. A mouse dendritic cell line, DC2.4 cells, and DCs that were generated from mouse bone marrow cells by culturing with granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-4 were each pretreated with AF for 2 hr, and then incubated with ovalbumin (OVA). After the 2-hr incubation, the DCs were fixed, and the amounts of OVA peptide-H-2Kb complexes were assessed using OVa-specific CD8+ T cells. AF inhibited MHC class I restricted presentation of exogenous OVA. This inhibitory activity of AF appeared to be due not only to the inhibition of the phagocytic activity of DCs, but also to the suppression of MHC molecule expression on DCs. AF also inhibited MHC class II-restricted presentation of exogenous OVA. These results show that AF exerts immunosuppressive activity at least in part by inhibiting MHC-restricted antigen presentation in professional antigen-presenting cells. PMID- 18409054 TI - Evaluation of the stability of aspirin in solid state by the programmed humidifying and non-isothermal experiments. AB - The influence of both moisture and heat on the stability of aspirin was investigated by a single pair of experiments, one with programmed humidity control and the other non-isothermal, rather than many standard isothermal studies, each at constant relative humidity. In experiments, we adopted the acid base back titration method to measure the content of aspirin in the presence of its degradation products. It was found that the degradation of aspirin could be expressed as ln[(c0-c)/c]=kt+D, where D was a lag time item not related to humidity and temperature. The relationship between the degradation rate constant k and humidity Hr) and temperature T could be described as Arrhenius equation multiplied by an exponential item of relative humidity: k = A . exp(mHr) . exp( (Ea/RT)), where A, Ea and m were the pre-exponential factor, observed activation energy, and a parameter related to humidity, respectively. The results obtained from the programmed humidifying and non-isothermal experiments, A=(1.09+/ 2.04)x10(12) h(-1), Ea=(93.5+/-2.2) kJ . mol(-1) and m=1.18+/-0.19, were comparable to those from isothermal studies at constant humidity, A=(1.71+/ 0.35)x10(12) h(-1), Ea=(94.9+/-0.7) kJ . mol(-1) and m=1.20+/-0.02. Since the programmed humidifying and non-isothermal experiments save time, labor and materials, it is suggested that the new experimental method can be used to investigate the stability of drugs unstable to both moisture and heat, instead of many classical isothermal experiments at constant humidity. PMID- 18409055 TI - Solid dispersions of dihydroartemisinin in polyvinylpyrrolidone. AB - In the present study the physicochemical characteristics of dihydroartemisinin, polyvinylpyrrolidone and their solid dispersions were evaluated at various proportions of drug and polyvinylpyrrolidone. These properties were investigated with X-ray diffraction, fourier transform infrared spectrophotometry, differential scanning calorimetry, equilibrium solubility at twenty five and thirty seven degree centigrade. X-ray diffraction analysis detected that dihydroartemisinin became more amorphous as drug carrier ratio was enhanced in solid dispersions. Fourier transform infrared spectra suggested that there was a hydrogen bonding interaction between dihydroartemisinin and polyvinylpyrrolidone in all solid dispersions. These interactions reflected the changes in crystalline structures of dihydroartemisinin. The thermal behavior of dihydroartemisinin was unusual as it exhibited melting exotherm instead of endotherm. In solid dispersions containing varying contents of polyvinylpyrrolidone, enthalpy change and peak area were enhanced while melting onset temperature decreased with increase in polyvinylpyrrolidone proportion. This was attributed to a solid-state interaction. Equilibrium solubility of dihydroartemisinin increased sixty-fold due to induction of polyvinylpyrrolidone. When this solubility was compared among thirty-seven and twenty five degree centigrade in solid dispersions, it was up to seven times more at higher temperature. Physicochemical characteristics of solid dispersions containing drug carrier ratio of one: nine prepared in acetonitrile, ethanol, methanol and tetrahydrofuran showed differences which indicated that properties of medium i.e. dielectric constant, dipole moment and structure, influenced the amount of amorphousness and related properties of dihydroartemisinin. PMID- 18409056 TI - Paroxetine hydrochloride controlled release POLYOX matrix tablets: screening of formulation variables using Plackett-Burman screening design. AB - The aim of the present study was to screen the effects of the formulation variables - POLYOX molecular weight (X1), the ratio of POLYOX/Avicel PH102 (X2) and the amount of POLYOX and Avicel PH102 (X3), hardness (X4), HPMCP amount (X5), Eudragit L100 amount (X6), and citric acid amount (X7) - on the paroxetine hydrochloride release from POLYOX matrix tablet using the Plackett-Burman screening design. Paroxetine hydrochloride matrix tablets were prepared according to a 7-factor-12-run statistical model and subjected to a 8-h dissolution study in Tris buffer at pH 7.5. The regression results showed that POLYOX molecular weight (X1) and POLYOX/Avicel PH102 ratio (X2) had significantly influence on the drug release mechanism and drug release rate as main effects. Hardness (X4) had an insignificant effect on the drug release mechanism but a significant effect on the drug release rate. On the other hand, HPMCP, Eudragit L100 and citric acid had an insignificant effect on the both responses. The information obtained by screening design study can be expected to be useful for further formulation studies. PMID- 18409057 TI - In vitro metabolic stability of moisture-sensitive rabeprazole in human liver microsomes and its modulation by pharmaceutical excipients. AB - A reliable method to assess in vitro metabolic stability of rabeprazole and its modulation by Generally Recognized As Safe (GRAS)-listed pharmaceutical excipients was established in human liver microsomes. The metabolic stability of rabeprazole decreased as a function of incubation time, resulting in the formation of thioether rabeprazole via nonenzymatic degradation and enzymatic metabolism. Buffer type was also a determining factor for the degree of both nonenzymatic degradation and enzymatic metabolism. The net extent of enzymatic drug metabolism, obtained by calculating the difference in drug degradation between a microsome-present reaction system and a microsome-free solution, was about 9.20 +/- 0.67% in phosphate buffer and 2.27 +/- 1.76% in Tris buffer, respectively. Rabeprazole exhibited first-order kinetics in microsome-free solution but showed non-linear kinetics in the microsome-present reaction system. The maximal velocity, Vmax, in phosphate buffer was 5.07 microg mL(-1) h(-1) and the Michaelis-Menten constant, Km, was 10.39 microg mL(-1) by computer-fitting to the classical Michaelis-Menten equation for pattern of time-dependent change in the substrate concentration. The intact drug and its thioether form were well resolved and successfully identified by HPLC chromatography and liquid chromatography mass spectroscopy (LC/MS). The metabolic stability of rabeprazole was also modulated by the presence of pharmaceutical excipients. Among the five pharmaceutical excipients tested, poloxamer 188 and Gelucire 44/14 had potentially inhibitory effects on rabeprazole metabolism in human liver microsomes (p < 0.05). A greater understanding of metabolic stability and its modulation by pharmaceutical excipients would be useful for optimizing the bioavailability of rabeprazole at the early formulation stages. PMID- 18409058 TI - Transport of PCBs with leachate water from the contaminated soil. AB - Contaminated soil was taken from the area around the damaged capacitor of an electrical transformer station in Zadar (Croatia) and a phytoremediation experimental field was constructed with lysimeters. The levels of PCBs in the leachate water samples were measured nearly 3 years. The experiment was conducted under natural climatic conditions, through hot summers and mild winters. Although the amounts of leachated PCBs from the polluted soil were relatively small, their concentrations in leachate water samples from 1,500 to 29,000 ng L(-1) substantially exceeded the maximum allowed concentration of the total PCBs in the fishponds of Croatia (1 ng L(-1)). PMID- 18409059 TI - Expression and activity of a probable toxin from Photorhabdus luminescens. AB - As an insect pathogen, Photorhabdus luminescens possesses an arsenal of toxins. Here we cloned and expressed a probable toxin from P. luminescens subsp. akhurstii YNd185, designated as Photorhabdus insecticidal toxin (Pit). The pit gene shares 94% nucleotide and 98% predicted amino acid sequence identity with plu1537, a predicted ORF from P. luminescens subsp. laumondii TT01 and 30% predicted amino acid sequence similarity to a fragment of a 13.6 kDa insecticidal crystal protein gene of Bacillus thuringiensis (Bt). The pit was expressed as a GST-Pit fusion protein in E. coli, most of which was insoluble and sequestered into inclusion bodies. The inclusion bodies were harvested and dissolved. The resultant protein was purified and the Pit was cleaved from the fusion protein by thrombin and purified from GST then used for bioassay. Pit killed Galleria mellonella (LD(50), 30 ng/larva) and Spodoptera litura (LD(50), 191 ng/larva) via hemocoel injection. Relative to a control that lacked toxin, Pit did not significantly increase mortality of S. litura and Helicoverpa armigera when introduced orally, but the treatment did inhibit growth of the insects. The present study demonstrated that Pit possessed insecticidal activity. PMID- 18409061 TI - Detection of human mammaglobin mRNA in serial peripheral blood samples from patients with non-metastatic breast cancer is not predictive of disease recurrence. AB - INTRODUCTION: Human mammaglobin (hMAM) mRNA is a sensitive and specific marker of breast cancer cells. We evaluated if hMAM mRNA detection in serial peripheral blood samples from non-metastatic breast cancer patients predicts for disease recurrence. METHODS: Patients scheduled for adjuvant or neoadjuvant chemotherapy were eligible. Serial blood samples were collected up to 5 years, the first before (neo)adjuvant chemotherapy. hMAM gene expression was analysed by RT-PCR. Specificity was evaluated in blood samples from healthy volunteers. A total of 321 patients were included. RESULTS: The incidence of pre-chemotherapy hMAM positive samples was similar in patients who latter experienced cancer recurrence (22.4%) and those who remained disease-free (17.9%; P = 0.46). Similarly, the mean number of positive follow-up samples was similar in both groups (0.15 +/- 0.22 and 0.13 +/- 013; P = 0.29). Furthermore, there was no difference in disease free (P = 0.63) or overall survival (P = 0.57) in patients with and without positive baseline samples or between patients whose follow-up samples were always hMAM negative and those with at least one positive sample. Multivariate survival analysis confirmed that hMAM mRNA detection before or after (neo)adjuvant chemotherapy was not predictive of recurrence. DISCUSSION: There is no evidence that hMAM mRNA detection at diagnosis or during follow-up predicts for breast cancer recurrence. PMID- 18409060 TI - The effect of thrombospondin-1 on breast cancer metastasis. AB - Thrombospondin-1 (TSP-1) has been proposed to have both pro-metastatic and anti metastatic properties. To elucidate its role in breast cancer metastasis, we compared tumor progression in the polyomavirus middle T antigen (Pyt) transgenic mouse and the TSP-1-null Pyt transgenic mouse. We characterized the tumors in these mice at 45, 60 and 90 days of age. Tumor size, areas of necrosis, macrophage infiltration, levels of active and total TGF-beta, vessel morphology, and lung and blood metastasis were measured in these mice. Mammary tumors were larger in the TSP-1-null mouse, and vessels were larger, but fewer in number in these tumors. The level of total TGF-beta was significantly higher in the Pyt tumors at 90 days of age. Importantly, significantly fewer metastases were observed in the lungs of the TSP-1-null/Pyt mouse. Primary Pyt tumor cells were more migratory than TSP-1-null Pyt tumor cells on collagen. Treatment of Pyt mice with recombinant proteins that contain the type-1 repeats of TSP-1 resulted in decreased primary tumor growth and metastasis. Sequences that are involved in CD36 binding and those required for TGF-beta activation mediated the inhibition of primary tumor growth. Thus, TSP-1 in the mammary tumor microenvironment inhibits angiogenesis and tumor growth, but promotes metastasis to the lung in the Pyt transgenic mouse. The ability of TSP-1 to support metastasis correlates with its ability to promote tumor cell migration. PMID- 18409062 TI - Recognizing acute hepatitis C in hemodialysis patients. PMID- 18409063 TI - An infantile case of Hinman syndrome with severe acute renal failure. AB - A 1-year-6-month-old Japanese girl with Hinman syndrome manifested urosepsis and severe obstructive nephropathy. Her voiding cystourethrogram (VCUG) revealed high grade vesicoureteral reflux with hydronephrosis; urodynamic study was compatible with detrusor-sphincter dyssynergia. She was treated conservatively, including clean intermittent catheterization. At 3 years old, bladder function had not improved, and estimated creatinine clearance was in the subnormal range. Hinman syndrome is a potential cause of acute and chronic renal failure in infancy. Taking account of the possibility of this condition in any neonates or infants who present urinary tract infection (UTI) appears to be necessary, since early recognition and proper management of this syndrome may prevent serious complications. PMID- 18409064 TI - Inhibitory effects of falconensins on 12-O-tetradecanoylphorbol-13-acetate induced inflammatory ear edema in mice. AB - Fifteen derivatives of falconensin were examined for their inhibitory activity against the induction of ear edema in mouse ear by application of 12-O tetradecanoylphorbol-13-acetate (TPA). It was demonstrated that azaphilonoid falconensins exert inhibitory effects against TPA-induced inflammation in the ears of mice to a comparable degree as moascorubrin and indomethacin, which are known to have antitumor-promoting and anti-inflammatory effects. All compounds tested, except monomethylmitorubrin, inhibited the inflammatory activity induced by TPA. PMID- 18409065 TI - Rearranged ent-kauranes from the stems of Tricalysia dubia and their biological activities. AB - From the EtOAc-soluble fraction of an MeOH extract of the stems of Tricalysia dubia, two new rearranged ent-kaurane derivatives, named tricalysiolides H and I, were isolated, together with five known rearranged ent-kauranes, i.e. tricalysiolides A-E, stigmast-4-en-6beta-ol-3-one, (+)-pinoresinol, scopoletin and syringaldehyde. Their structures were elucidated from spectroscopic evidence, and their cytotoxicity toward KB cells and P-gp inhibitory activity were assayed. PMID- 18409066 TI - Antioxidant constituents in the dayflower (Commelina communis L.) and their alpha glucosidase-inhibitory activity. AB - The dayflower, Commelina communis L., contains 1-deoxynojirimycin (DNJ) and (2R,3R,4R,5R)2,5-bis(hydroxymethyl)-3,4-dihydroxypyrrolidine (DMDP), potent alpha glucosidase inhibitors. The extracts and powder of this herb are important food materials for prophylaxis against type 2 diabetes. Eleven flavonoid glycosides as antioxidants, isoquercitrin, isorhamnetin-3-O-rutinoside, isorhamnetin-3-O-beta-D glucoside, glucoluteolin, chrysoriol-7-O-beta-D-glucoside, orientin, vitexin, isoorientin, isovitexin, swertisin, and flavocommelin, were identified from the aerial parts of C. communis. Their antioxidant activities were measured using in vitro assays employing the 1,1-diphenyl-2-picrylhydrazyl radical- and superoxide radical-scavenging assays. The results showed that glucoluteolin, orientin, isoorientin, and isoquercitrin are the predominant antioxidants in this herb. Moreover, isoquercitrin, isorhamnetine-3-O-rutinoside, vitexin, and swertisin inhibited the activity of alpha-glucosidase from rat intestine. PMID- 18409067 TI - Anaerobic degradation of p-xylene in sediment-free sulfate-reducing enrichment culture. AB - Anaerobic degradation of p-xylene was studied with sulfate-reducing enrichment culture. The enrichment culture was established with sediment-free sulfate reducing consortium on crude oil. The crude oil-degrading consortium prepared with marine sediment revealed that toluene, and xylenes among the fraction of alkylbenzene in the crude oil were consumed during the incubation. The PCR denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene for the p-xylene degrading sulfate-reducing enrichment culture showed the presence of the single dominant DGGE band pXy-K-13 coupled with p-xylene consumption and sulfide production. Sequence analysis of the DGGE band revealed a close relationship between DGGE band pXy-K-13 and the previously described marine sulfate-reducing strain oXyS1 (similarity value, 99%), which grow anaerobically with o-xylene. These results suggest that microorganism corresponding to pXy-K-13 is an important sulfate-reducing bacterium to degrade p-xylene in the enrichment culture. PMID- 18409068 TI - Src inhibitors in early breast cancer: a methodology, feasibility and variability study. AB - Early clinical trials of anticancer agents may be enriched by robust biomarkers of activity. Surrogate measures used in trials of cytotoxic agents, such as tumor size regression, may not be informative when investigating targeted agents that act principally to inhibit invasion or proliferation. This study aimed to determine the validity of invasion-related biomarkers of activity for AZD0530, a potent Src inhibitor currently in clinical development. Focal adhesion kinase (FAK) and paxillin are downstream phosphorylation substrates of Src and mediate tumor cell adhesion and invasiveness. These were therefore selected as biologically relevant markers of Src inhibition. Early breast cancer was chosen as a model as multiple samples can be collected during standard treatment and there is an intervening period in which experimental intervention can be applied. Tumor tissue was collected from diagnostic core biopsies and subsequent surgical tumor excision samples in 29 women with early breast cancer attending a single center. Protein levels were assessed quantitatively by Luminex and qualitatively by immunohistochemistry. AZD0530 inhibited tumor growth in a manner independent of dose and inhibited phosphorylation of FAK and paxillin in a dose-dependent manner in a Calu-6 xenograft model. In the clinical study, agreement of within visit and also of between-visit measurements was high and the estimated number of patients required to detect a drug effect would be low enough to allow use of these markers as endpoints in future dose selection studies. PMID- 18409069 TI - Changes in quality of life among breast cancer patients three years post diagnosis. AB - OBJECTIVES: To evaluate changes in quality of life (QOL) and identify medical and socio-demographic predictors of QOL among breast cancer survivors 3 years after diagnosis. METHODS: Between April 2002 and March 2004 2,232 women with breast cancer were recruited into the Shanghai Breast Cancer Survival Study, a population-based cohort study, approximately 6 months after cancer diagnosis. QOL was evaluated at 6 and 36 months post-diagnosis using the General Quality of Life Inventory. Multiple linear regression models were used to evaluate changes in QOL and their associations with socio-demographic and medical factors. RESULTS: In general, breast cancer patients showed significant improvement over time in the physical, psychological, and social well-being domains, as well as overall QOL. However, QOL scores in several facets did not improve or even deteriorated, including increased negative feelings, reduced social support and interpersonal relationships, and worsened financial situation and living environment. Age at diagnosis was inversely associated with QOL changes in physical and psychological well-being and overall QOL scores. Compared with those who were disease-free, patients with a recurrence of disease had significantly poorer QOL scores across all well-being domains and almost all facets. Patients who received radiotherapy experienced significant improvements in physical and social well-being and overall QOL. Mixed ER/PR status was associated with improved scores in physical and psychological well-being and overall QOL. Education, body mass index, Charlson comorbidity score, TNM stage, type of surgery, chemotherapy, and immunotherapy were only associated with changes in certain well-being domains and/or facets, but not overall QOL. Tamoxifen use was not associated with changes in QOL outcomes. CONCLUSIONS: Our study provides valuable information for developing strategies/interventions for improving the QOL of breast cancer patients. PMID- 18409070 TI - Association of single nucleotide polymorphisms (SNPs) in TNF-LTA locus with breast cancer risk in Indian population. AB - PURPOSE: Cytokine milieu of tumor microenvironment affects tumorigenesis in breast cancer. The aim of the present study was to investigate the potential association of functional single nucleotide polymorphisms (SNPs) in TNF-LTA locus with breast cancer. METHODS: The study included 127 individuals comprising 40 breast cancer cases (35 sporadic & 5 familial) and 87 individuals of high risk group (with family history of breast cancer) along with 150 healthy controls. PCR RFLP was employed to analyze TNFA promoter polymorphisms at -238 G/A, -308 G/A, 857 C/T, -863 C/A and -1031 T/C along with +252 A/G SNP in LTA. The results were further confirmed by direct sequencing. RESULTS: Significant association was established for TNFA -308 G/A and LTA +252 A/G polymorphisms with breast cancer versus controls (P < 0.0001; OR, 9.53; 95% CI, 4.11-22.13; P (c) < 0.001) and high risk group versus controls (P < 0.0001; OR, 8.27; 95% CI, 4.28-16.0; P (c) < 0.001) respectively. GGACCT haplotype was found to be positively associated with breast cancer (P < 0.0001; OR, 12.17; 95% CI = 5.12-28.92; P (c) < 0.001) and high risk group (P, 0.03; OR, 2.95; 95% CI, 1.20-7.26; P (c), 0.005) in relation to controls. While GGGCCT haplotype was significantly related with high risk group in comparison to cancer (P, 0.0002; OR, 5.71; 95% CI, 2.18-14.99; P (c), 0.003) and controls (P, 0.0002; OR, 2.48; 95% CI, 1.55-3.96; P (c), 0.003). CONCLUSION: TNF-LTA locus could serve as an important biomarker for breast cancer predisposition in Indian population. PMID- 18409073 TI - [Drugs for postoperative analgesia: routine and new aspects: Part 2: opioids, ketamine and gabapentinoids]. AB - In part 1 of this review, perioperative aspects of the use of non-opioids (acetaminophene, dipyrone, traditional NSAR, coxibs) were discussed. In part 2 the perioperative aspects of opioids (weak opioids: tramadol, tilidine with naloxone, strong opioids: morphine, piritramide, oxycodone, hydromorphone, fentanyl, methadone, buprenorphine) and coanalgesics (gabapentinoids; ketamine) will now be presented. The main aim of the review is to describe the use, risks and cost of some substances to facilitate the differential indication. New aspects concerning the use of gabapentinoids and ketamine are discussed. PMID- 18409071 TI - Activation of ErbB3, EGFR and Erk is essential for growth of human breast cancer cell lines with acquired resistance to fulvestrant. AB - Seven fulvestrant resistant cell lines derived from the estrogen receptor alpha positive MCF-7 human breast cancer cell line were used to investigate the importance of epidermal growth factor receptor (ErbB1-4) signaling. We found an increase in mRNA expression of EGFR and the ErbB3/ErbB4 ligand heregulin2 (hrg2) and a decrease of ErbB4 in all resistant cell lines. Western analyses confirmed the upregulation of EGFR and hrg2 and the downregulation of ErbB4. Elevated activation of EGFR and ErbB3 was seen in all resistant cell lines and the ErbB3 activation occurred by an autocrine mechanism. ErbB4 activation was observed only in the parental MCF-7 cells. The downstream kinases pAkt and pErk were increased in five of seven and in all seven resistant cell lines, respectively. Treatment with the EGFR inhibitor gefitinib preferentially inhibited growth and reduced the S phase fraction in the resistant cell lines concomitant with inhibition of Erk and unaltered Akt activation. In concert, inhibition of Erk with U0126 preferentially reduced growth of resistant cell lines. Treatment with ErbB3 neutralizing antibodies inhibited ErbB3 activation and resulted in a modest but statistically significant growth inhibition of two resistant cell lines. These data indicate that ligand activated ErbB3 and EGFR, and Erk signaling play important roles in fulvestrant resistant cell growth. Furthermore, the decreased level of ErbB4 in resistant cells may facilitate heterodimerization of ErbB3 with EGFR and ErbB2. Our data support that a concerted action against EGFR, ErbB2 and ErbB3 may be required to obtain complete growth suppression of fulvestrant resistant cells. PMID- 18409074 TI - [Expression patterns of non-viral transfection with GFP in the organ of Corti in vitro and in vivo. Gene therapy of the inner ear with non-viral vectors]. AB - BACKGROUND: Diseases of the inner ear such as presbycusis, tinnitus, sudden hearing loss, and vertigo affect many patients, but so far there are no specific therapy options. Gene therapy might become a potential modality of treatment. Viral vectors are standard in animal models to date. Future considerations, however, call for a further evaluation of non-viral transfection methods. MATERIAL AND METHODS: The non-viral transfection agents Metafectene, Superfect, Effectene, and Mirus TransIT were incubated with a plasmid coding for GFP. In vivo, the plasmid-agent mix was injected via the membrane of the round window, and 48 h later the inner ear was perfused, harvested, decalcified, and histologically evaluated for GFP expression. RESULTS: Cationic lipids (Metafectene) and dendrimers (Superfect) were able to transfect cells in the area of the organ of Corti and lead to GFP expression. The polyamine (Mirus TransIT) did show expression of GFP in the area of Rosenthal's canal and in the area of the inner hair cell. The combination of a non-liposomal lipid with a DNA condensing component (Effectene) did not show transfection of the organ of Corti. In the area of the spiral ganglia cells, GFP expression was seen with all the transfection agents. CONCLUSIONS: Non-viral transfection agents are able to introduce a reporter gene in cells of the inner ear in vitro and in vivo. There are, however, differences in the efficiency of the transfection. They might be an alternative in gene therapy of the inner ear. Further investigations to elucidate their potential are needed. PMID- 18409075 TI - Laboratory evaluation of a phytosteroid compound of mature leaves of Day Jasmine (Solanaceae: Solanales) against larvae of Culex quinquefasciatus (Diptera: Culicidae) and nontarget organisms. AB - Crude mature leaves extract of Day Jasmine, Cestrum diurnum (Solanaceae: Solanales), was investigated for larvicidal activities against Culex quinquefasciatus, the vector of human filariasis. All the graded concentrations (0.1%, 0.2%, 0.5%, 1%, 1.5%, 2%, 2.5%, and 3% v/v) showed significant (P < 0.05) larval mortality, and results of regression equations revealed that the mortality rates were positively correlated with the concentrations of the extract (R (2) close to 1). LC(50), LC(95), and LC(99) values were calculated at different time intervals, and the lowest value was recorded at the 72-h bioassay for third instar larvae. Significant changes in the larval mortality (F < 0.05) was also recorded between the periods of exposure and between instars during t test and single analysis of variance analysis. No mortality was noticed for nontarget organisms, such as Oreochromis niloticus niloticus and Chironomid larvae within 72 h of postexposure to 1%, 2%, and 3% crude plant extract under the laboratory condition. Qualitative and chromatographic analysis of the crude extract of C. diurnum revealed the presence of many bioactive phytochemicals. The bioassay experiment with the third-instar larval form established that the presence of a steroid compound (R (f), 0.67) was responsible for mosquitocidal activity. PMID- 18409076 TI - Leukocytosis is linked to thrombosis at diagnosis, while JAK2 V617F mutation is associated with thrombosis during the course of essential thrombocythemia. PMID- 18409077 TI - Multiple brain infarctions induced by imatinib mesylate in a patient with clonal eosinophilia. PMID- 18409078 TI - Primary non-Hodgkin lymphomas in the small and large intestine: clinicopathological characteristics and management of 40 patients. AB - To investigate the clinicopathological characteristics and optimal treatment modalities of primary non-Hodgkin lymphoma (NHL) in the small and large intestine. Forty patients with primary NHL in the small and large intestine were studied retrospectively. All cases were reclassified according to the World Health Organization (WHO) classification of lymphoma in 2001. Fourteen patients had primary disease in the small intestine, which were all of B-cell origin with diffuse large B-cell lymphoma (DLBCL) diagnosed in 5 of 14 (35.7%) patients and mucosa-associated lymphoid tissue (MALT) lymphoma in 8 of 14 (57.1%) patients. Ileum was the most commonly involved site (8 of 14 patients, 57.1%), followed by jejunum (2 of 14 patients, 14.3%) and duodenum (1 of 14 patients, 7.1%). Twenty five patients had primary colorectal lymphoma, with B-cell origin accounting for 92.0% and T-cell origin for 8.0% of these patients. The ileocaecal region has the highest involved rate (13 of 25 patients, 52.0%), followed by colon (7 of 25 patients, 28.0%) and rectum (3 of 25 patients, 12.0%). Compared with surgery alone, post-operation chemotherapy or chemoradiotherapy can significantly improve DLBCL patients' event-free survival (EFS). However, no post-operation treatment modality can improve OS or EFS for patients with MALT lymphoma. B-cell lymphoma is the most common pathological type of intestinal lymphomas. Chemotherapy containing treatment modality is an effective way to improve intestinal lymphoma patients' EFS, especially for those with DLBCL subtype. PMID- 18409079 TI - Increased incidence of interstitial pneumonia by CHOP combined with rituximab. AB - Several authors have reported interstitial pneumonia (IP) during rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) therapy, while others have encountered Pneumocystis jirovecii pneumonia during rituximab combined bi-weekly CHOP. Herein, we report that 13 of 90 (14%) patients developed IP during R-CHOP therapy, compared with none of 105 patients treated with CHOP alone as a historical control. There were no differences in baseline data between patients undergoing the two therapies. Among R-CHOP-treated patients, serum beta D-glucan was increased in 8 of 12 (75%) IP patients compared with none of 30 non IP patients examined. In five IP patients who underwent sputum evaluation, two were positive for P. jirovecii by the polymerase chain reaction and another two were positive for Candida albicans. No other organisms were detected as causative pathogens. Treatment with steroids, sulfamethoxazole-trimethoprim (ST), and antifungals was effective. Our results suggest that R-CHOP raises the incidence of IP, possibly through increasing the susceptibility to P. jirovecii and fungal infection. The need for prophylactic antifungals and ST during R-CHOP should be evaluated by randomized controlled trials. PMID- 18409080 TI - Reserpine can confer stress tolerance and lifespan extension in the nematode C. elegans. AB - Though the lifespan extension mechanism is partly understood from C. elegans to mice, a viable pharmacological intervention is not yet feasible. Here, we report that reserpine largely known as an antipsychotic-antihypertensive drug, can extend C. elegans lifespan. Chronic reserpine treatment from embryo stage or young adults extends the C. elegans lifespan robustly at 25 degrees C. Most importantly, the reserpine treated long lived worms are active (locomotion and pharyngeal pumping) for a long time thereby conferring high quality throughout life. Reserpine mediated lifespan extension is independent of the daf-16 pathway and partly requires serotonin. Reserpine treatment makes the worms highly thermotolerant. Thus, in addition to its known function, reserpine is able to provide stress tolerance and lifespan extension in C. elegans. PMID- 18409081 TI - Single nucleotide polymorphism (SNP) at the GHR gene and its associations with chicken growth and fat deposition traits. AB - 1. The growth hormone receptor (GHR) plays crucial roles on chicken growth and metabolism. 2. The full cDNA of the chicken GHR gene was scanned for single nucleotide polymorphisms (SNP) by means of denaturing high-performance liquid chromatography (DHPLC). Three SNP, C6540334T, C6542011T and G6631778A, were genotyped in a F(2) designed full-sib resource population to analyse their associations with chicken growth and fat deposition traits. 3. Fifty-five SNP and two other variations were identified in the 8908 bp region of the GHR gene. Among the 55 SNP, 10 were located in coding exons (6 resulted in changes of amino acids) and 45 were in non-coding regions (introns, 5'UTR and 3'UTR). The nucleotide diversity (theta), corrected for sample size of chicken GHR gene, is 1.45 x 10(-3). Fourteen PCR-RFLP markers were developed in the chicken GHR gene. 4. The G6631778A was associated with body weight at 63 d (BW63), dressed weight (DW) and subcutaneous fat thickness (SFT), BW35 and BW49 (P < 0.01) as well as hatch weight (HW) and BW42 in the male population. However, G6631778A was only associated with BW28 in the female population. G rather than A was dominant for chicken growth and fat deposition. Haplotypes based on the three SNP were associated with BW21, BW70, BW77 and SFT, BW7, BW35, BW42, BW49 and BW56 in males, and associated with BW7 and BW14 in females. For growth in males, the H2 and H6 haplotypes had positive and negative effects, respectively; meanwhile H6 was predominant for fat deposition. PMID- 18409083 TI - Effect of the naked neck gene on carcase composition and immunocompetence in chicken. AB - 1. An experiment was conducted to study the effect of naked neck gene (Na) on carcase measurements and some immunological responses of chickens. 2. The presence of naked neck gene in a single or double state increased body weight. 3. The naked neck genotypes (NaNa or Nana) exhibited higher relative weight of dressed carcase, drumstick and breast muscles compared to normally feathered individuals (nana). The proportion of abdominal fat was decreased in both naked neck genotypes compared with normally feathered ones. 4. A higher (non-specific) cell-mediated immune response was evoked in Na gene bearing birds. 5. Higher concentrations of antibodies occurred after injection of sheep red blood cells into naked neck birds, particularly at 14 d post-secondary injection. 6. The NaNa birds had a higher heterophil/lymphocyte ratio than Nana and nana siblings. PMID- 18409082 TI - Divergent selection for shape of growth curve in Japanese quail. 4. Carcase composition and thyroid hormones. AB - 1. Changes in the relative weights of carcase, abdominal fat, breast and leg muscles, and plasma thyroid hormone concentrations occurring during the first 6 weeks of postnatal growth were analysed in males of HG and LG lines divergently selected for high and low relative body weight (BW) gain between 11 and 28 d of age, respectively, and constant adult BW. 2. The second week of postnatal life was a critical age at which the HG males exhibited a relatively faster growth in comparison to their LG counterparts and permanently exceeded LG males in the percentage by weight of carcase, breast and leg muscle. A higher production of muscle tissues was associated with lower accumulation of abdominal fat before sexual maturity. 3. In general, the plasma T(3) level of HG quail exceeded that of LG quail. Nevertheless, significant differences were found only at 14, 21 and 28 d of age, that is, in the period during which the highest inter-line differences in relative growth rate were noted. Also the T(3)/T(4) ratio followed a similar trend while plasma T(4) level showed no clear and consistent inter-line differences. 4. The results suggest that the selection for the shape of the growth curve, like the selection for body fat, modifies the carcase quality owing to shortening/prolongation of the acceleration growth phase. Individuals with a short acceleration phase of the growth curve are characterised by low carcase quality during the fattening period. PMID- 18409084 TI - Immune responses induced by DNA vaccines encoding Newcastle virus haemagglutinin and/or fusion proteins in maternal antibody-positive commercial broiler chicken. AB - 1. The immune responses induced by recombinant plasmids containing Newcastle disease virus (NDV) F (pVAX.nd.f) or HN (pcDNA.nd.hn) genes separately or in combination in bi-cistronic (pIRES.nd.hn.f) constructs were evaluated in maternal antibody-positive commercial chicks. 2. Immunofluorescence and immunoperoxidase tests demonstrated the expression of both F and HN proteins in Vero cells. Real time PCR analysis revealed the expression of HN and/or F genes in muscle, peripheral blood mononuclear cells (PBMC), spleen and liver after immunisation. 3. Chicks inoculated intramuscularly thrice (two booster doses) with pVAX.nd.f and pcDNA.nd.hn did not develop detectable haemagglutination inhibiting (HI) antibodies. In contrast, an increase in a NDV-specific cell-mediated immune response was demonstrated. 4. After challenge with virulent NDV, chicks immunised with the recombinant plasmids as well as those in control groups succumbed to Newcastle disease. 5. Based on these results, it is concluded that DNA vaccines containing HN and/or F genes fail to protect commercial chicks, possibly due to interference from maternal antibodies. PMID- 18409085 TI - Aspects of meat and eating quality of broiler chickens reared under standard, maize-fed, free-range or organic systems. AB - 1. The effects of rearing chickens using standard, maize-fed, free-range or organic production systems on meat quality and sensory characteristics were evaluated. The standard system used either Ross or Cobb birds with a slaughter age of approximately 40 d, the other systems in the trial used Hubbard birds with considerably older slaughter ages, up to 72 d in the case of organic systems. 2. Paired breast fillets from 120 birds, 30 from each rearing system, were used. The meat quality variables, ultimate pH, colour coordinates L*, a*, b* and water holding capacity were measured and taste panel assessments were made using 8 point category scales of texture, juiciness, abnormal flavour, flavour liking and overall flavour. 3. There were significant differences between rearing systems, with fillet muscles from birds grown under the standard system having a higher ultimate pH. Differences were also seen in colour with fillets from birds reared under a standard system having a smaller hue angle than those grown using the maize-fed system which had the highest. 4. Fillets from birds reared in the standard system were rated by the taste panel as more tender and juicy. There were no significant differences in chicken flavour. Based on hedonic assessments of flavour liking and overall liking, by a small panel of assessors, meat from birds produced in the standard system was most preferred and that from organic systems the least preferred. Meat from free-range and maize-fed systems was intermediate in preference. This result reveals a trend, but does not infer consumer acceptance. PMID- 18409086 TI - Comparative antibacterial activity of avian egg white protein extracts. AB - 1. Egg white proteins from the eggs of domestic chicken (Gallus gallus), turkey (Meleagris gallopavo), duck (Anas platyrhynchos) and goose (Anser anser) were analysed in order to compare the antimicrobial activity of these products. 2. Albumen from each species was sampled and analysed by SDS-PAGE and Western blotting. Antimicrobial activity and lysozyme activity were measured. 3. Ovotransferrin and ovalbumin were identified in all species while c-type lysozyme was present in chicken, turkey and duck egg white samples, but not in goose. 4. Galliformes appear to possess albumens with greater antimicrobial activity than those of the Anseriformes. This can be attributed to higher concentrations of ovotransferrin and the broad acting c-type lysozyme. PMID- 18409087 TI - Antimicrobial activity of cuticle and outer eggshell protein extracts from three species of domestic birds. AB - 1. The eggshell cuticle is the proteinaceous outermost layer of the eggshell which regulates water exchange and protects against entry of micro-organisms. In this study, we investigated the hypothesis that the cuticle may also reduce microbial contamination by providing a chemical defence. 2. Outer eggshell and cuticle protein was extracted from domestic chicken (Gallus gallus), duck (Anas platyrhynchos) and goose (Anser anser) eggs by HCl and urea treatment, respectively. Antimicrobial activity of the extracts against Gram-positive and Gram-negative bacteria was evaluated. 3. C-type lysozyme, ovotransferrin and ovocalyxin-32 were identified in all extracts by Western blotting. All extracts from all species demonstrated lysozyme enzymatic activity. Immobilised c-type lysozyme retained some enzymatic activity. Protein extracts demonstrated activity against Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis suggesting the action of antimicrobial proteins in addition to lysozyme. 4. The results suggest that the antimicrobial outer eggshell and cuticle proteins present in a number of avian species may be a mechanism which enhances avian reproductive success. PMID- 18409088 TI - Effects of dietary phytase on performance and nutrient metabolism in chickens. AB - 1. A broiler growth study was conducted to compare the effect of different concentrations of an Escherichia coli-derived phytase on performance, apparent metabolisable energy (AME), nitrogen (N), amino acid and mineral metabolisability, sialic acid excretion and villus morphology when fed to broiler chickens. 2. Female Ross 308 broilers (480) were reared in floor pens from 0 to 28 d of age. All birds were fed on nutritionally complete starter (0 to 21 d of age) and grower diets (21 to 28 d of age) with the exception that they were low in P (28 and 23 g/kg available P, respectively). These maize-soy diets were supplemented with 0, 250, 500 or 2500 phytase units (FTU)/kg feed. 3. Between 21 and 28 d of age, two birds from each floor pen were selected, and each pair placed in one of 32 metabolism cages (two birds per cage). Feed intake was recorded and excreta collected for the last 2 d of the feeding period, and AME, N, amino acid and mineral metabolisability coefficients and endogenous losses were determined following a total collection procedure. 4. Feed intake and weight gain increased in a linear manner in response to phytase dose, with an average increase of approximately 11.7 and 13.5%, respectively, compared with chickens fed on the low-P diet. Birds given diets with 2500 FTU weighed 6.6% more and had a 2.4% higher feed conversion efficiency (FCE) than those fed on diets containing 500 FTU. 5. Enzyme supplementation increased the intake of AME and metabolisable N by 10.3 and 3.9%, respectively, principally through increases in feed intake. Birds given enzyme-supplemented diets also improved their intake of metabolisable amino acids and P by approximately 14 and 12.4%, respectively, compared with birds fed on the control diet. Enzyme supplementation did not affect ileal villus morphometry of the birds. PMID- 18409089 TI - Production and carcase traits in broiler chickens given diets supplemented with inorganic trivalent chromium and an organic acid blend. AB - 1. The study was conducted to ascertain the effects of supplemental organic acids and chromium (Cr) on production and carcase traits of broiler chickens. 2. A total of 120 1-d-old broiler chicks were divided into 4 treatment groups in a 2 x 2 factorial design (each treatment group contained 6 replicates with 5 birds per replicate). 3. The diets were supplemented with an organic acid blend containing ortho-phosphoric, formic and propionic acid and calcium propionate (1 g/kg diet) and inorganic trivalent chromium (Cr(3+)) as chromic chloride hexahydrate (0.5 mg/kg diet) either independently or together as a combination for 35 d. 4. Individual supplementation of organic acids and Cr(3+) and their combination significantly improved the food conversion ratio, hot and dressed carcase weight and weight of the wholesale cuts compared to the control group of birds. 5. Organic acids, either independently or along with Cr(3+), increased total accretion of ash in carcase. Protein accretion was improved by dietary Cr(3+) and organic acid supplementation compared to the control group and a further improvement in this regard was observed when Cr(3+) and organic acid were supplemented together. Across the treatment groups meat fat content and fat accretion were lower in birds receiving dietary Cr(3+) supplementation. 6. Circulatory Cr(3+) and meat Cr(3+) concentration increased compared to the other treatment groups when Cr(3+) was supplemented to the birds. 7. It was concluded that, instead of individual supplementation, a combination of Cr(3+) and organic acids may improve the production and carcase traits of broilers more effectively presumably because of an additive effect. PMID- 18409090 TI - Polysaccharidase preparations added to a wheat-based diet: effects on performance and digestive parameters of broiler chickens held at three different locations. AB - 1. Three experiments of 39 d were simultaneously conducted to assess the efficacy of three carbohydrase combinations on performance, apparent nutrient digestibility, intestinal morphology and digestive organ relative weights of chickens fed on a wheat-based diet. 2. Experiment 1 was performed under laboratory conditions, with the birds housed in cages. In experiment 2, the birds were placed in floor pens built in a commercial farm, and raised with their contemporaries at the farm. In experiment 3, broilers were reared under industrial management circumstances in commercial farms. A two-phase feeding programme was used: starter from 1 to 21 d of age and finisher from 22 to 39 d. The basal diet without supplementation (BD) and three enzyme preparations (BDF, BDP and BDR) were evaluated. 3. No differences in growth traits were observed among treatments in experiments 1 and 2. 4. When overall growing period was evaluated, all enzyme combinations improved apparent DM ileal digestibility of feed in the three experiments, but significant differences were only found in experiments 2 and 3. For this same period, an increase in apparent CP ileal digestibility of broiler chickens fed supplemented diets against the control group was found in experiment 3. Moreover, a significant greater apparent ileal fat digestibility in the enzyme treatments compared with the control group was shown when broilers were raised under industrial conditions (experiment 3). 5. Enzyme combinations had no effect on villus heights and crypt depths of broilers in experiment 1, whereas, in experiment 2, a greater villus height was found in BDF and BDP treatments than in control birds. 6. There were no differences in digestive organ relative weights among dietary treatments except for pancreas in experiment 1. PMID- 18409091 TI - Comparative digestibility of amino acids of maize, sorghum, finger millet and pearl millet in cockerels and Japanese quails. AB - 1. The objective of the present study was to determine the apparent digestibilities of amino acids of maize, low tannin sorghum, pearl millet and finger millet in adult cockerels and Japanese quails. 2. Adult White Leghorn cockerels (25 weeks of age) and adult Japanese quails (15 weeks of age) were used to determine the apparent digestibilities of amino acids in cereal grains. 3. The digestibilities of amino acids in sorghum determined with quails were similar to cockerels, with the exception of histidine, which was significantly lower in quails. 4. In finger millet, the digestibilities of methionine and leucine were significantly higher, while that of histidine and cystine were significantly lower in adult quails than in adult cockerels. 5. The digestibilities of pearl millet amino acids were similar in quails and cockerels, except for that of histidine, which was significantly lower in quails. 6. Almost all the amino acids in maize, except histidine, had significantly higher digestibility values in quails than in cockerels. Histidine digestibility of all the 4 cereal grains was significantly lower in quails than in cockerels. 7. The amino acid digestibility values for certain cereal grains determined with cockerels do not support their use in formulating practical quail diets. The amino acid digestibility values of maize were underestimated by using cockerels while that of finger millet was overestimated. PMID- 18409092 TI - The influence of caecectomy on amino acid availability of three feedstuffs for ganders. AB - 1. Experiments were conducted to assess the influence of caecectomy on amino acid availability (AAA) of three feedstuffs for goose. 2. Nine caecectomised and 9 intact Yangzhou ganders, 24 weeks old, were used in these experiments. Fish meal, soybean meal and cottonseed meal were used as the sole source of protein. The endogenous amino acid (AA) losses were evaluated by a nitrogen (N)-free diet method. The influence of caecectomy on apparent amino acid availability (AAAA) in fish meal, soybean meal and cottonseed meal was assessed in experiment 1 and true amino acid availability (TAAA) of three protein diets was determined in experiment 2. 3. Results showed that, in the soybean meal and cottonseed meal, the AAAA and TAAA of most AA determined by the intact ganders were higher than in the caecectomised ganders; in the fish meal, the AAAA and TAAA of most AA determined by the intact ganders were lower than in the caecectomised ganders. 4. Results of the present study suggest that the effect of caecectomy on AAA in geese was dependent on the feedstuff assayed, and it was better to use caecectomised poultry for AAA assessment. PMID- 18409093 TI - Effects of mannanoligosaccharide in broiler chicken diets on growth performance, energy utilisation, nutrient digestibility and intestinal microflora. AB - 1. A study was undertaken to evaluate the effects of mannanoligosaccharide (MOS, Bio-MOS, Alltech Inc.) on the growth performance, energy utilisation, nutrient digestibility and intestinal microflora of birds given a sorghum-wheat based diet. Two MOS levels (1 and 2 g/kg) were included in the diet. 2. Inclusion of MOS at both levels in the diet improved the apparent metabolisable energy (AME) values of the diet. However, these effects were not as pronounced as those of zinc bacitracin (ZnB) treatment. Dietary ZnB also significantly improved the net energy value of the diet. No significant differences between the different levels of MOS were noticed in the growth performance, AME and net energy values of the diet. Compared to the negative control, inclusion of 2 g/kg MOS tended to improve feed conversion efficiency (FCE) in the starter phase. 3. Dietary MOS did not affect the apparent total tract digestibility of nutrients compared to the negative control. In contrast, ZnB significantly improved the protein digestibility and tended to increase the starch digestibility. The addition of MOS reduced the concentration of arabinose in the soluble non-starch polysaccharides (NSP) fraction in the excreta of birds; whereas, the concentrations of individual sugars in the insoluble NSP and free sugar fractions were increased by ZnB. 4. A decrease in the populations of lactobacilli and coliforms in the ileal and caecal lumen was observed for MOS and ZnB treatments. Correspondingly, pH and microbial fermentation in the gut was altered. The addition of MOS tended to reduce the coliform load at the gut mucosa. 5. Results from the current study suggest that MOS can improve the apparent energy utilisation of the diet and tend to improve FCE of birds in the first three posthatch weeks, which may be partly related to the modulatory effects of MOS on the gut microflora. PMID- 18409094 TI - Effect of reducing dietary mineral content on growth performance, water intake, excreta dry matter content and blood parameters of broilers. AB - 1. The objective of this study was to investigate whether it is possible to reduce phosphorus (P) excretion by feeding a lower level of dietary P than that currently recommended, without compromising welfare, and whether this is sensitive to the level of dietary calcium (Ca). 2. A randomised complete block design was used with a total of 224 1-d-old male chicks of a commercial genotype in two batches. Chicks had a 10-d adaptation period. After this, they were moved to cages and allocated to one of 4 dietary treatments (4 and 3 birds per cage in batch 1 and 2, respectively). A standard level of Ca and P (8.3 and 6.6 g/kg, respectively) was compared with three lower levels of Ca (7.3, 6.3 and 5.3 g/kg) all at the same reduced level of P (5.5 g/kg). Records were made of live weight of birds on an individual basis and feed and water consumption on a cage basis. Every 7 d, the dry matter (DM) content of a sample of mixed excreta from each cage was measured. Birds were slaughtered and samples taken for analysis of plasma mineral content, retention of N, Ca and P and tibia bone strength. 3. The initial reduction in dietary mineral level did not adversely affect performance or bone strength. However, decreasing dietary Ca level below 7.3 g/kg, at constant P, led to reduced feed consumption and bone strength. Dietary treatment had no significant overall effect on water intake, water : feed ratio, excreta DM content, Ca and P retention or metabolisability of DM. There was no significant correlation between mean plasma Ca or P level and DM content of rectal digesta. 4. In conclusion, reducing dietary P from 6.6 to 5.5 g/kg to reduce potential environmental impact of broiler production had an adverse effect on bird welfare when the level of Ca was reduced below 7.3 g/kg, as a result of decreased feed intake in the early growth period and reduced skeletal strength. PMID- 18409095 TI - Effects of different feed additives alone or in combination on broiler performance, gut microflora and ileal histology. AB - 1. The objectives of this experiment were to investigate the effects of different xylanases, alone or in combination with different organic acid and oligosaccharide sources, on bird performance, gut microflora and ileal histology. 2. Birds were given a diet based on a commercial formulation, which was split into 8 batches. Batch 1 contained the antibiotic growth promoter Avilamycin and acted as the positive control. To batch 2 the enzyme Allzyme PT was added and to batch 3 Allzyme PT was added with the organic acid and oligosaccharide mixture Avimos. To batch 4, Allzyme PT was added with the oligosaccharide mixture Biomos. To batch 5, yeast extract 2012 was added with the organic acid mixture Gustor and the enzyme xylanase XP20. To batch 6, yeast extract 2012 and feed acidifier Gustor were added as before, with the enzyme Avizyme 1300. Batches 7 and 8 both acted as negative experimental controls, with no added growth promoters. 3. A total of 64 birds were housed in individual wire cages in each of three consecutive experimental replicates (24 birds/treatment). Birds were fed ad libitum from 7 to 28 d and a 7-d excreta collection was carried out to determine apparent metabolisable energy (AME) content. 4. At 28 d, the birds were killed and viscosity of jejunal digesta supernatant and gizzard weight were determined. Samples were taken from the crop, ileum and caecum and analysed for viable presumptive lactic acid bacteria and coliforms. The overall microbial flora was determined using denaturing gradient gel electrophoresis of 16S ribosomal DNA followed by DNA sequence analysis in order to assign amplicons to a bacterial species. Ileal sections were also collected for histological analysis. 5. Total live weight gain (12%) and gain:feed (9%) were significantly improved for all diets containing additives, compared to the negative control diets. All diets containing xylanases gave significantly lower in vivo viscosity values than the positive and negative controls. Diet treatment significantly affected viable coliform numbers in the ileum and also viable lactobacilli in the ileum and caecum. A substantial proportion of the bacteria present in the GI tract (40%) belong to unknown species. No effects of diet treatment on histological measurements were observed in this study. 6. All the additive combinations studied were at least as effective as the antibiotic growth promoter and the results for Allzyme PT suggest that xylanase alone is as effective as any of the combinations studied. PMID- 18409096 TI - Dietary conjugated linoleic acid improves antioxidant capacity in broiler chicks. AB - 1. The influence of dietary conjugated linoleic acid (CLA) on the antioxidant status in the absence or presence of endotoxin exposure was studied with male broiler chicks. 2. In experiment 1, a total of 240 1-d-old broilers were allotted into 4 dietary groups (0, 2.5, 5.0 or 10.0 g pure CLA/kg) to study the influence of CLA on growth performance and antioxidant defence systems. The results showed that growth performance was not altered by 42 d of CLA consumption. Increased total superoxide dismutase (TSOD) activities in liver, serum and muscle were observed in chicks given 10.0 g CLA/kg diet. Dietary CLA at 10.0 g/kg also markedly elevated liver catalase (CAT) activity. Malondialdehyde (MDA), a marker of lipid peroxidation, decreased in liver, serum and muscle in chicks given 5.0 and 10.0 g CLA/kg diet. 3. In experiment 2, a total of 120 1-d-old broilers were fed on a control diet (without CLA) or 10.0 g CLA/kg diet. Half of the birds fed on each diet were injected intraperitoneally with 0.25 mg/kg body weight of Salmonella enteritidis lipopolysaccharide (LPS) at 16, 18 and 20 d of age. Decreased glutathione peroxidase (GSH-Px), TSOD activity and increased ceruloplasmin and MDA concentrations were seen in the challenged chicks. Dietary CLA prevented the loss of body weight gain and feed conversion ratio of chicks followed repeated endotoxin exposure. CLA partially inhibited the increase of serum ceruloplasmin and MDA at 17 and 21 d of age and notably suppressed the decrease of serum TSOD activity at 21 d of age. 4. These results suggested that dietary CLA enhances the activity of antioxidant enzymes including TSOD and CAT. Supplementation of CLA has been shown to ameliorate the antioxidant balance and performance of chicks during oxidative stress. PMID- 18409097 TI - Effects of high CO2 level during early incubation and late incubation in ovo dexamethasone injection on perinatal embryonic parameters and post-hatch growth of broilers. AB - 1. A total of 1200 Cobb broiler breeder eggs were incubated under either standard conditions (C group) or high CO(2) levels (CO(2) group) during the first 10 d of incubation. The CO(2) level of the CO(2) incubator was attained gradually by a natural build-up of CO(2) due to air-tight closure of the incubator. From d 10 of incubation onwards, all eggs were incubated under standard incubation conditions. At d 18 of incubation, the eggs of both incubation groups (CO(2) and C group) were either injected with water-soluble dexamethasone (DEXA group) or with saline (0.9% NaCl; saline group) or were not injected (control). 2. Body weights, plasma hormonal concentrations (T(3), T(4) and corticosteroid) and glucose concentrations were measured regularly during the perinatal (at IP, internal pipping) and post-hatch period (at 1, 2, 4 and 6 weeks post-hatch). Additionally, hatchability and pattern of embryonic mortality were determined after hatch. 3. The results showed that high CO(2) levels during the first 10 d of incubation or dexamethasone injection at d 18 of incubation decreased embryo mortality, mainly because of a reduction in embryo malpositioning. However, a combination of a CO(2) incubation and a dexamethasone injection led to an increase in embryo mortality and therefore a decrease in hatchability. 4. Although dexamethasone injection at embryonic d 18 decreased body weight in the second week of the rearing period and CO(2) incubation increased body weight during the first 2 weeks of the rearing period, no consistent effect of both the incubation and injection treatments on body weight at slaughter age was observed. PMID- 18409098 TI - Arm coordination in elite backstroke swimmers. AB - In this study, we assessed arm coordination in the backstroke over increasing speeds by adapting the index of coordination originally used in the front crawl. Fourteen elite male backstroke swimmers swam four trials of 25 m at the speeds corresponding to the 400-m, 200-m, 100-m, and 50-m events. The six phases of the arm stroke were identified by video analysis and then used to calculate the index of coordination, which corresponded to the time between the propulsive phases of the two arms. With increases in speed, the elite swimmers increased the stroke rate, the relative duration of their arm pull, and their index of coordination, and decreased the distance per stroke (P < 0.05). Arm coordination was always in catch-up (index of coordination of -12.9%) because the alternating body-roll and the small shoulder flexibility did not allow the opposition or superposition coordination seen in the front crawl. This new method also quantified the relative duration of the hand's lag time at the thigh, which did not change ( approximately 2%) with increasing speed for the elite swimmers. The index of coordination enables coaches to assess mistakes in backstroke coordination, particularly in the hand's lag time at the thigh. PMID- 18409099 TI - Heart rate and blood lactate concentration of male road-race motorcyclists. AB - Although motorcycling performance strongly depends on the characteristics of the motorcycles and capabilities of the riders, little information is available on the physiological profiles of riders. The aim of this study was to evaluate the physical load of official international men's road-race motorcycling competitions. Data were obtained from 34 male riders during the 2005 European Road-Race Motorcycling Championship (categories classified by size of engine: 125 GP, 250 GP, and 600 cc) during free practices, qualifying sessions, and official races. Participants' heart rates were recorded and blood lactate concentrations determined. During races, heart rates were most often above 90% of maximum heart rate (frequency of occurrence: 125 GP = 92.9%, s = 5.3; 250 GP = 93.6%, s = 7.3; 600 cc = 93.2%, s = 10.2). The heart rate distribution during riding showed main effects between phases of competition, engine sizes, and different portions of the race (P < 0.001). No difference was observed between riders on and not on the podium at the end of the race. Peak blood lactate concentrations after the qualifying sessions (5.2 mmol . l(-1), s = 1.2) and official races (6.0 mmol . l( 1), s = 2.1) were higher (P < 0.001) than at baseline. The present results show that road-race motorcycling imposes a high load on the riders, who should possess adequate fitness to maintain high-speed rides and minimize the effects of fatigue during competition. PMID- 18409100 TI - An objective method for depicting team performance in elite professional rugby union. AB - Using a two-study approach, we examined a methodology for objectively depicting team performance and form in a professional rugby union side. Study 1 developed standardized indicators to examine a team's performance in a single match relative to their previous matches over a domestic season via a performance report and form chart. This resulted in standardized performance indicators that provided instant and coherent feedback on the team's performance relative to previous standards. Study 2 then utilized this methodology to compare a match between two professional sides, played the following season, to assess the extent to which the performance by one team affected that of the other. Comparison of the two teams' performances, for the match they played against each other, identified a drop in relative performance (against previous standards) for both teams on the same performance indicators. This appeared to be due to the match being a close, hard-fought contest particularly in the tackle and around the gain line. The findings of the two studies suggest an accurate and viable methodology for depicting team performance that is superior to the global measures previously adopted. Presenting teams' current standardized performances (i.e. form) on a single visual scale also has utility for coaches within an applied setting. Future research should further investigate the effect of one team's performance on another through both individual and team outcomes. PMID- 18409101 TI - ATP and phosphocreatine utilization in single human muscle fibres during the development of maximal power output at elevated muscle temperatures. AB - In this study, we examined the effect of muscle temperature (Tm) on adenosine triphosphate (ATP) and phosphocreatine utilization in single muscle fibres during the development of maximal power output in humans. Six male participants performed a 6-s maximal sprint on a friction-braked cycle ergometer under both normal (Tm = 34.3 degrees C, s = 0.6) and elevated (T(m) = 37.3 degrees C, s = 0.2) muscle temperature conditions. During the elevated condition, muscle temperature of the legs was raised, passively, by hot water immersion followed by wrapping in electrically heated blankets. Muscle biopsies were taken from the vastus lateralis before and immediately after exercise. Freeze-dried single fibres were dissected, characterized according to myosin heavy chain composition, and analysed for ATP and phosphocreatine content. Single fibres were classified as: type I, IIA, IIAX25 (1 - 25% IIX isoform), IIAX50 (26 - 50% IIX), IIAX75 (51 75% IIX), or IIAX100 (76 - 100% IIX). Maximal power output and pedal rate were both greater (P < 0.05) during the elevated condition by 258 W (s = 110) and 22 rev . min(-1) (s = 6), respectively. In both conditions, phosphocreatine content decreased significantly in all fibre types, with a greater decrease during the elevated condition in type IIA fibres (P < 0.01). Adenosine triphosphate content was also reduced to a greater (P < 0.01) extent in type IIA fibres during the elevated condition. The results of the present study indicate that after passive elevation of muscle temperature, there was a greater decrease in ATP and phosphocreatine content in type IIA fibres than in the normal trial, which contributed to the higher maximal power output. PMID- 18409102 TI - Validity of the VmaxST portable metabolic measurement system. AB - The aim of this study was to assess and compare the validity of the portable VmaxST telemetry metabolic measurement device with that of a standard measurement system (Vmax29). Thirty asymptomatic, moderately active males provided written, informed consent and completed two maximal graded treadmill exercise tests (Bruce) using the VmaxST and the Vmax29 metabolic measurement systems. Tests were performed in random order on separate days to obtain peak values for time to exhaustion, heart rate, systolic and diastolic blood pressure, oxygen consumption (VO2), carbon dioxide production (VCO2), ventilation (VE), and respiratory exchange ratio (RER). Multivariate analysis of variance revealed no significant main effect (P = 0.88) between the two systems across any variable, suggesting similar measurement capabilities between the two systems. Linear regression analyses revealed moderate to high coefficients of determination for VO2 (r2 = 0.99), VCO2 (r2 = 0.99), VE r2 = 0.99), and RER (r2 = 0.89). Furthermore, Bland Altman analyses demonstrated that the VmaxST yielded similar values to the Vmax29, suggesting good agreement between the two systems. Agreement was confirmed when the differences between the methods resulted in a small range as identified by the 95% limits of agreement. Findings from the current study confirm that the VmaxST is a valid device for measuring metabolic and physiological variables during exercise within a controlled laboratory setting. PMID- 18409103 TI - Goal orientations, motivational climate, and prosocial and antisocial behaviour in youth football: exploring their temporal stability and reciprocal relationships. AB - In this study, we examined the temporal stability and reciprocal relationships among task and ego orientation, task- and ego-involving climates, and prosocial and antisocial behaviour in youth football. Male (n = 156) and female (n = 24) footballers (mean age 14.1 years, s = 1.8) completed questionnaires towards the beginning and end of a regular season. Questionnaires measured goal orientation, perceived motivational climate, and frequency of prosocial and antisocial behaviours. Structural equation modelling indicated moderate covariance stability between the beginning and end of the season. Subsequent analyses revealed a significant decrease only in perceptions of task-involving climate. In the cross lagged analyses, prosocial behaviour at the beginning of the season positively predicted task-involving climate at the end of the season. Antisocial behaviour at the beginning of the season positively predicted both ego orientation and ego involving climate at the end of the season and a reciprocal relationship was revealed whereby ego orientation at the beginning of the season positively predicted antisocial behaviour at the end of the season. Task orientation at the beginning of the season negatively predicted ego-involving climate at the end of the season. All cross-lagged relationships were weak. This exploratory study offers limited support for bi-directional relationships between personal, environmental, and behavioural variables but provides useful insight into the covariance stability, change, and interrelationships between motivational and moral constructs over a competitive season. PMID- 18409104 TI - The effect of rowing headgear on forced convective heat loss and radiant heat gain on a thermal manikin headform. AB - Both radiant and forced convective heat flow were measured for a prototype rowing headgear and white and black cotton caps. The measurements were performed on a thermal manikin headform at a wind speed of 4.0 m . s(-1) (s = 0.1) in a climate chamber at 22.0 degrees C (s = 0.05), with and without radiant heat flow from a heat lamp, coming from either directly above (90 degrees ) or from above at an angle of 55 degrees . The effects of hair were studied by repeating selected measurements with a wig. All headgear reduced the radiant heat gain compared with the nude headform: about 80% for the caps and 95% for the prototype rowing headgear (P < 0.01). Forced convective heat loss was reduced more by the caps (36%) than by the prototype rowing headgear (9%) (P < 0.01). The radiant heat gain contributed maximally 13% to the net heat transfer, with or without headgear, showing that forced convective heat loss is the dominant heat transfer parameter under the chosen conditions. The results of the headgear - wig combinations were qualitatively similar, with lower absolute heat transfer. PMID- 18409105 TI - Comparison of physiological responses to graded exercise test performance in outrigger canoeing. AB - The aim of this study was to establish a graded exercise test protocol for determining the peak physiological responses of female outrigger canoeists. Seventeen trained female outrigger canoeists completed two outrigger ergometer graded exercise test protocols in random order: (1) 25 W power output for 2 min increasing by 7.5 W every minute until exhaustion; and (2) 25 W power output for 2 min increasing by 15 W every 2 min to exhaustion. Heart rate and power output were recorded every 15 s. Expired air was collected continuously and sampled for analysis at 15-s intervals, while blood lactate concentration was measured immediately after and 3, 5, and 7 min after exercise. The peak physiological and performance variables examined included peak oxygen uptake (VO2peak), minute ventilation, tidal volume, ventilatory thresholds 1 and 2, respiratory rate, respiratory exchange ratio, heart rate, blood lactate concentration, power output, performance time, and time to VO2peak. There were no significant differences in peak physiological responses, ventilatory thresholds or performance variables between the two graded exercise test protocols. Despite no significant differences between protocols, due to the large limits of agreement evident between protocols for the peak physiological responses, it is recommended that the same protocol be used for all comparison testing to minimize intra individual variability of results. PMID- 18409106 TI - Contextual influences on baseball ball-strike decisions in umpires, players, and controls. AB - Baseball umpires, players, and control participants with no baseball experience were asked to call balls and strikes for video clips. In a basic judgement task, umpires and players were significantly better at calling pitches than controls. In a direct information task, borderline pitches were presented following clips of definite balls and definite strikes. Participants called target pitches closer to the strike end of the scale when viewed after definite balls than when they followed definite strikes. Similarly, when borderline pitches were shown in different pitch counts, participants called pitches more towards the strike end of the scale when there were three balls in the count (3-0, 3-2). These findings indicate that the standard for evaluation changes based on the context in which stimuli are processed. Moreover, the strength of the contextual factors is illustrated in that the effects were shown in observers with and without experience in the task. Overall, however, umpires had a greater tendency to call strikes, indicating that they may use a norm of "hastening the game". PMID- 18409107 TI - The effect of biofeedback training on affective regulation and simulated car racing performance: a multiple case study analysis. AB - The foundation of this study was based on an idiosyncratic concept, which uses probabilistic determinations (Kamata, Tenenbaum, & Hanin, 2002) to verify the utility and effectiveness of a biofeedback intervention by manipulating affective performance states in a race-car simulator. Nine males completed five separate time-trials of a simulated racing task and were then randomly assigned to one of three arousal regulation treatment conditions: (1) optimal, (2) poor, and (3) attention control. Following the biofeedback intervention, participants underwent another series of race trials to determine the effectiveness of the arousal regulation intervention. The results indicated that there were relative similarities in the strength and direction of the perceived and physiological states between the participants; however, the subtle details of the participants' unique performance zones and the probability of achieving each zone were revealed to be unique among the participants. The results also indicated that: (a) the biofeedback manipulation resulted in the expected changes for each participant, and (b) there were some large individual differences among the participants, necessitating the idiosyncratic approach. Limitations and future directions are also addressed. PMID- 18409108 TI - Centre of pressure sway characteristics during static one-legged stance of athletes from different sports. AB - The frequency of one-legged stance and two-legged stance differs considerably among sports. We therefore expect the balance ability of athletes from different sports to vary. This study compared the sway characteristics during a static one legged stance of soccer players, basketball players, swimmers, and non-athletes. The centre of pressure sway during one-legged stance of ten male participants representing each of the four groups was measured using a stabilometer. Centre of pressure sway was assessed by four sway factors: sway velocity, anterior posterior sway, horizontal sway, and high-frequency sway. None of the four groups of participants showed significant differences in body sway between standing on the dominant leg and standing on the non-dominant leg. The soccer players had more high-frequency sway and less anterior-posterior sway and horizontal sway than the basketball players, swimmers, and non-athletes. These results suggest that soccer players have superior ability to maintain a stable one-legged stance. Further study is required to determine how much of the superior balance ability in soccer players is innate and how much is developed through training, as well as to determine the relationship between balance ability and playing performance. PMID- 18409109 TI - Measures of cardiac autonomic control in women with differing volumes of physical activity. AB - In this study, we investigated heart rate variability in women who exercise regularly, but with differing volumes of physical activity. Forty women participated and were allocated to either a "high" or "moderate" activity group. Heart rate as R-R intervals was recorded during supine rest and analysed using time domain, frequency domain, and Poincare plot techniques. All time domain and Poincare analysis measures revealed significant differences in heart rate variability between the "high" and "moderate" activity groups. Frequency domain analysis also revealed a significant difference between the two groups in absolute measures of high-frequency and low-frequency power, but not between measures such as the low-frequency/high-frequency ratio, proposed to represent the sympathetic/parasympathetic balance of cardiac control. The results demonstrate that there is a significant difference in heart rate variability between women who engage in "high" compared with "moderate" volumes of physical activity, which may have implications for cardiac autonomic health. PMID- 18409112 TI - Antifouling activity of the sponge metabolite agelasine D and synthesised analogs on Balanus improvisus. AB - This study reports a screening study for antifouling (AF) activity of the natural compound agelasine D isolated from marine sponges of the genus Agelas and 20 synthesised analogs of agelasines and agelasimines. Agelasine D, together with two of the analogs, ie AV1003A and AKB695, displayed a strong inhibitory effect on settlement of Balanus improvisus cypris larvae. Agelasine D had an EC50 value of 0.11 microM while the two analogs AV1033A and AKB695 had EC50 values of 0.23 and 0.3 microM, respectively. None of these three compounds affected larval mortality as was the case with several of the analogs tested. Moreover, the effect of AV1033A and AKB695 was reversible. When cyprids after 24 h exposure to the compounds were transferred to fresh seawater, the settlement frequency compared with the controls was completely recovered. The properties of the agelasine D analogs AV1003A and AKB695 make them highly attractive candidates as AF agents in future marine coatings. PMID- 18409111 TI - Quantification of lipids and protein in thin biofilms by fluorescence staining. AB - The efficiency of removing unwanted biofilm from surfaces in industrial water systems was examined by fluorescence microscopy and image analysis. A quantitative assay for in situ determination of biofilm components was developed and tested on thin biofilms grown in reactors as well as real biofilms sampled from a fish processing factory. Different fluorescent dyes for in situ detection of protein, lipid and total organic matter were tested. It was possible to determine the approximate amounts, concentrations and coverage of the different components by correlating the fluorescent intensity of the biofilm components to standard solutions immobilised as a biofilm. The quantification methods were evaluated as a strategy for determining the efficiency of different disinfection/cleaning procedures, showing that quantification of these biofilm components was fast and reliable for optimisation of cleaning in place procedures. However, the approach also showed that bacterial cells, as investigated by culture-independent procedures, were killed but not removed by most disinfection procedures tested, potentially leading to surfaces which are easily recolonised. PMID- 18409113 TI - Alpha-galactosides: antinutritional factors or functional ingredients? AB - This review focuses on updated information about alpha-galactosides, their chemical structure, biosynthesis, plant physiological functions, occurrence in foods, positive and negative physiological effects in animals, changes during food processing, and their potential application as prebiotics in the food industry. Although alpha-galactosides are considered as the main flatus-causing factors, they are also involved in several important functions during plant and seed development and beneficially stimulate the growth and activity of living bifidobacteria and lactobacilli in the human colon. We focus here also on legumes as a source of this kind of prebiotics as potential health promoters. PMID- 18409114 TI - Healthy indexes in public health practice and research: a review. AB - The scientific community has long been interested in the overall quality of diets owing to the fact that it is important for each individual's health through a healthy, varied, and balanced diet. Much research has been conducted on methods used to measure dietary quality. These studies led to the determination of numerous indices, some very simple and some much more complex. Indices that examine diets for several attributes concurrently are able to provide a measure of overall diet quality that is not possible when only single nutrients or food groups are examined. Using these indices, diet quality may be evaluated in relation to nutrient adequacy, compliance with dietary guidelines, or nutrition recommendations, association with risk of chronic diseases, or mortality, and used to assess interventions. However, there are several factors that should be taken into consideration for the design of an index, like the purpose of its use, as well as its simplicity in daily practice. The general purpose of an index is to synthesize a large amount of information into a single useful indicator. The purpose of this review was to present and to critically review the most commonly used dietary indices, and how they reflect various aspects of diet quality. The majority of these indices fail to recognize the various inter-relationships between their components, as well as their accuracy for estimating specific health outcomes. Thus, the development of weighted dietary indices that adequately assess a dietary pattern and its relationship to the burden of a disease is considered essential. PMID- 18409115 TI - The initial freezing temperature of foods at high pressure. AB - The Pure water (P,T)-phase diagram is known in the form of empirical equations or tables from nearly a century as a result of Bridgman's work. However, few data are available on other aqueous systems probably due to the difficulty of high pressure measurements. As an alternative, six approaches are presented here to obtain the food phase diagrams in the range of pressure 0.1-210 MPa. Both empirical and theoretical methods are described including the use of an artificial neural network (ANN). Experimental freezing points obtained at the laboratory of the authors and from literature are statistically compared to the calculated ones. About 400 independent freezing data points of aqueous solutions, gels, and foods are analysed. A polynomial equation is the most accurate and simple method to describe the entire melting curve. The ANN is the most versatile model, as only one model allows the calculation of the initial freezing point of all the aqueous systems considered. Robinson and Stokes' equation is successfully extended to the high pressures domain with an average prediction error of 0.4 degrees C. The choice of one approach over the others depends mainly on the availability of experimental data, the accuracy required and the intended use for the calculated data. PMID- 18409116 TI - Phase separation in biopolymer gels: a low- to high-solid exploration of structural morphology and functionality. AB - Phase separation in protein and polysaccharide gels remains one of the basic tools of achieving the required structural properties and textural profile in food product formulations. As ever, the industrialist is faced with the challenge of innovation in an increasingly competitive market in terms of ingredient cost, product added-value, and expectations of a healthy life-style to mention but a few. It appears, however, that a gap persists between the fundamental knowledge and a direct application to food related concepts with a growing need for scientific input. Furthermore, within the context of materials science, there is a tendency to examine research findings in either low- or high-solid systems without considering synergistic insights/benefits to contemporary needs, spanning the full range of relevant time-, length-, and concentration scales. This review highlights the latest attempts made to utilize and further develop fundamental protocols from the advanced synthetic polymer research as a source of inspiration for contemporary bio-related applications in low- and intermediate-solid composite gels. Then, it takes advantage of this school of thought to "force a passage" through the phase topology and molecular dynamics of binary biopolymer mixtures at high levels of co-solute. It is hoped that these phenomenological and fundamental tools should be able to bridge the divide in the analysis of the two "types" of composite materials (from low to high solids) thus dealing effectively with the specific and often intricate problems of their science and applications. PMID- 18409117 TI - Results of inspections in health hazard industries in a region of the state of Washington. AB - Results from state of Washington Occupational Safety and Health Administration (OSHA) health inspections were reviewed to characterize the level of control that existed at the work sites, identify common problems, and assess the success of targeting programs in finding and correcting uncontrolled health hazards. Data were obtained from 170 inspections conducted by industrial hygienists in a four county area in 12 industries, including 10 that were targeted. Inspection data were accessed through the use of electronic databases and inspection reports then summarized by the following industries: bathtub refinishing, carbide tool and saw sharpening, fibercement siding installation, furniture refinishing, health care clinics, janitorial floor waxing service, landscaping, lawn maintenance, tree service, road construction, stone countertop fabrication, truck bed lining, warehouse and cold storage, and wood floor finishing. Targeted health hazards included worker exposure to airborne contaminants, noise, and bloodborne pathogens typical of the industry. Method and effectiveness of control of health hazards were evaluated by counting work sites with violations associated with engineering control, personal protective equipment, hearing conservation, or training. Results are presented by industry for the number and percentage of work sites that failed to provide required protection. Poor control of health hazards was generally found across all inspected industries. Follow-up inspections and self-reports of abatement found that more than 85% were able to successfully control the hazards and abate the violations. The results are further discussed as they relate to methods of control and risk and identifying existing and emerging high health hazard industries. Based on employment data and the poor hazard control that was found, most if not all the industries can be described as high health hazard, small-employer industries. The results can be used for the planning of interventions in other regions and industries. PMID- 18409118 TI - [Case report: severe hypothermia in a newborn infant - challenges in preclinical emergency medicine]. AB - On the basis of a case report the prehospital management of a newborn child with deep accidental hypothermia (22oC) is discussed. The child was found in a garbage can. The continuous resuscitation during the transport into the clinic is done in an incubator and the child survives without neurologic damages. The used measures of the resuscitation are discussed on the basis of the therapy. PMID- 18409119 TI - [Non-invasive ventilation in prehospital emergency medicine]. AB - Non-invasive ventilation has not yet been established in prehospital emergency medicine. This most likely due to missing technical prerequisites. Meanwhile emergency ventilators feasible for prehospital NIV are available. Recognizing the pathophysiology of acute respiratory insufficiency, treatment with NIV is superior in comparison to treatment with oxygen and medication only. The advantages of NIV may lead to reduced morbidity and mortality as long as attention is paid to possible contraindications. PMID- 18409120 TI - [Non-invasive ventilation in prehospital emergency medicine - contra]. AB - The non-invasive ventilation (NIV) has been practiced successfully for years in the clinical routine. Now this therapeutic option should be transferred in the pre hospital emergency service. But there are many risks, they must be mentioned: As a matter of fact an effective clinical therapy can not be brought to the streets easily - even if there are good and well-tried conservative therapies and just at last just a few patients. PMID- 18409121 TI - [Anesthesia and obesity - perioperative management of the obese patient]. AB - The prevalence of obesity is increasing and a growing number of people with overweight present for surgery. For preoperative evaluation, the patients have to be checked for obesity-related diseases. The assurance whether the equipment can stand the increased stress needs a thorough check up. Although there is no general tendency for a higher rate of difficult intubation or increased rates of aspiration, there is a need of particular consideration. Cardiovascular function as well as tolerance to surgical stress is diminished. Pulmonary function is deterioated and impaired for at least 48 hours after surgery, which also applies to renal, hepatic function and to coagulation and glucose hemostasis. The pharmacokinetics of most anaesthetics is altered in view of initial distribution, elimination and duration of effects. Obese people without significant impairments are suitable for ambulatory surgery. PMID- 18409122 TI - [Therapy without improved prognosis?]]. PMID- 18409123 TI - [Acute renal failure - how and when to treat?]. AB - Acute renal failure is a common condition in intensive care units. The negative impact of acute renal failure on mortality has been demonstrated in recent studies. All critically ill patients should be regarded as a high risk population for renal failure. The optimization of intravasal fluid status and mean arterial pressure are preventive strategies in these patients. The use of nephrotoxic drugs (including radiocontrast media) should be avoided if possible. In cases of established acute renal failure today therapeutic strategies are still limited to best supportive care. The use of diuretics can facilitate fluid balance, however they seem to have an adverse effect on excretional renal function. A number of patients with acute renal failure need extracorporal renal support. Overload of potassium or fluids, severe acidosis, uremic pericarditis or uremic encephalopathy are urgent indications for the start of renal replacement therapy. Small randomized trials give some evidence that an early start of renal replacement therapy may be beneficial in critically ill patients. In this patient group renal replacement therapy should be considered when serum urea concentrations exceed 100mg/dl and/or when early signs of indications mentioned above are present. Large randomized multicenter trials have shown that a favourable effect on mortality can only be achieved when renal replacement therapy is supplied with a sufficient dose. Daily hemodialysis or continuous hemofiltration with a filtrate volume of 35ml/kg/h is regarded as a standard of care. There is still controversy whether continuous hemofiltration is superior to intermittent hemodialysis. Large meta-analyses could not show a difference in mortality with either one of the two therapy options. PMID- 18409124 TI - [Acute renal failure - anticoagulation in continuous renal replacement therapy]. AB - Anticoagulation plays a pivotal role for the efficacy of continuous renal replacement therapy. The equilibrium between avoiding bleeding complications and keeping the system open considering patients} multiple diseases demands an individual and dynamic reasoned anticoagulation regime. For patients at risk of bleeding citrate anticoagulation has shown high efficacy and lesser bleeding complications than the standard heparin or other alternative anticoagulants such as regional protamin-heparin-, prostaglandin-heparin-anticoagulation or no anticoagulation. PMID- 18409125 TI - [Heparin induced thrombocytopenia and anticoagulation in renal replacemant therapy]. AB - The decision for an anticoagulant for renal replacement therapy (RRT) in patients with acute renal failure and heparin-induced thrombocytopenia (HIT) has to be made carefully. Based on results from the literature argatroban is favoured in patients without hepatic dysfunction, referring to its short halftime and easy feasable monitoring. In the case of coexsisting hepatic disorder, danaparoid provides a safe alternative therapy. However, long halftime and the difficult elimination of the substance are unfavourable. Lepirudin represents another possible anticoagulant therapy. Bleeding complications and monitoring of the ecarin clotting time imposes limitations. Experiences with bivalirudin, fondaparinux and prostaglandines are limited and future trials will have to determine the significance of their application in RRT in HIT patients. Furthermore it has to be proven whether the combination of alternative anticoagulants with citrate prolongates circuit halftime of CVVH. PMID- 18409126 TI - [Reorganization of the interdisciplinary emergency unit at the university clinic of Gottingen]. AB - Configuration of the interdisciplinary emergency unit within the university clinic of Gottingen was successfully reorganized during the past two years. All emergencies except traumatologic, gynecologic and pediatric emergencies are treated within this functional unit which is guided by the center of internal medicine. It is organized in a three shift operation manner over a period of 24 hours. Due to a close interdisciplinary collaboration between different departments patients receive optimal diagnostic and therapeutic treatment within a short period of time. To improve processes within the emergency department a series of measures were taken including the -establishment of an intermediate care unit for unstable patients, setting up of special diagnostic and therapeutic units for the acute coronary syndrome as well as stroke, implementation of standardized clinical pathways, establishment of an electronic data processing network in close communication with all diagnostic entities, introduction of a quality assurance system and reduction of medical costs. Reorganization measures lead to a substantial optimization and acceleration of emergency proceedings and thus, provides optimal patient care around the clock. In addition, medical costs could clearly be reduced at the interface between preclinical and clinical emergency medicine. PMID- 18409127 TI - [German research network on neuropathic pain: working on future therapy]. AB - Founded in 2002, the German Research Network on Neuropathic Pain includes the major institutions, authorities and medical disciplines engaged in therapy and research on Neuropathic pain. Together they work on a better understanding of the pathophysiology, the development of new therapies and the appropriate usage of established interventions for the treatment as well as the prevention of neuropathic pain. The network's central project is the establishment of a data bank, currently including standardized information of more than 1500 patients with neuropathic pain regarding socio-economic, psycho-social, psychological data and the results of quantitative sensory testing (QST), a standardized protocol implemented by the network. PMID- 18409128 TI - [Improvement of registration accuracy for navigated-control drill in mastoidectomy (autopilot)]. AB - BACKGROUND: The goal of this study is the improvement of the surgical accuracy of a navigate-controlled drill for mastoidectomy in a lab test. METHODS: For lab tests an artificial model of the temporal bone with color-coded injury identification of the facial nerve (solution of 0.5 mm) was used. Two different registration methods were examined: (group 1) navigation bow with 4 integrated markers at the upper jaw; (group 2) landmark registration with 4 titanium micro screws. An optical navigation system was used. The targets were illustrated by 3 titanium screws within the range of the planum mastoideum. The accuracy of the navigate-controlled drill in drilling the planned cavity were evaluated at 20 temporal bone models. The measurement of the registration accuracy was evaluated by deviation between the target screw and the calculated position in the navigation system. The evaluation of the resulted cavities was done by 5 senior surgeons with the help of the microscope. RESULTS: The registration accuracy shows a maximum deviation between the real position and the calculated position of 1,73 MM in group of 1 and 0.93 MM in group 2. In group 1 the nerve was hurt in 5/20 cases and a maximum deviation of - 1.5 mm (Std 0.25 mm) (drilled beyond the nerve) was measured. In group 2 the nerve was not hurt, a maximum deviation of 0.5 mm (too early stopped before the nerve) was measured. CONCLUSIONS: Significantly better results of the registration and drilling accuracy show up in group group 2. Thus the preconditions for clinical use are fulfilled. PMID- 18409129 TI - [Quality of life in children with obstructive sleeping disorder after tonsillectomy, tonsillotomy or adenotomy]. AB - BACKGROUND: To study the effectiveness of adenotomy (AT), adenotonsillectomy (ATE) and adenotonsillotomy (ATT) with respect to the quality of life improvement in children with obstructive sleep disorders (OSD), OSA 18 survey and Brouillette score were used. METHODS: Prospectively, 92 children with an age ranging from 2 to 6 years with OSD underwent AT, ATE or ATT at the University hospital of Ulm (Germany), respectively. 30 age-matched children served as controls. Caregivers were requested to complete the Brouillette score and the OSA 18 survey, which are validated instruments for detecting symptoms and quality of life change in children with OSDs, at the initial office visit prior to surgery (BS/1 and OSA 18/1) and 7 - 14 months after surgery (BS/2 and OSA 18/2). RESULTS: The mean total score of the OSA 18 survey for AT, ATE and ATT was reduced significantly after these operations (p < 0.001). Similar results were also observed in the Brouillette score (p < 0.002). The improvement of life quality in the AT group was less effective than in the ATE and ATT group. CONCLUSION: ATE and ATT have about the same effectiveness to improve life quality for children with OSD. Children after ATE or ATT have a comparable life quality as healthy children (Mann Whitney U-test; p approximately 0.15), while children who only received AT have a slightly lower life quality after the operation. In summary, ATT was recommend. PMID- 18409140 TI - Canine and feline dirofilariasis: life cycle, pathophysiology, and diagnosis. AB - Infection with the intravascular parasite Dirofilaria immitis is an increasingly recognized problem in domestic dogs and cats. Heartworm infection is preventable; however, once an animal is infected, heartworm disease and potentially life threatening complications can develop. An understanding of the heartworm life cycle and transmission season, the limitations of testing methods, and the clinical signs of heartworm disease is necessary to provide clients with appropriate information regarding prevention and treatment of heartworm infection in their pets. PMID- 18409141 TI - Canine and feline dirofilariasis: prophylaxis, treatment, and complications of treatment. AB - Several agents are available for the prevention of heartworm infection. Melarsomine is up to 96% efficacious (after two doses) as a heartworm adulticide in infected dogs. However, treatment of dogs infected with D. immitis can be expensive, and adulticide therapy in patients with moderate to severe heartworm disease can be associated with life-threatening complications. Patients with clinical signs associated with pneumonitis benefit from short-term therapy with antiinflammatory doses of corticosteroids before and after adulticide treatment. Strict cage rest for 4 to 6 weeks is mandatory after adulticide administration. Microfilaricide therapy is indicated for microfilaremic patients. The prognosis is good in patients with mild to moderate infection, fair to guarded in severe cases, and poor to grave (even with treatment) in patients with caval syndrome, severe pulmonary thromboembolism, or congestive heart failure. PMID- 18409142 TI - Understanding behavior: the multispecies household. AB - Although pet owners frequently have more than one species of pet, information on how to ensure a peaceful coexistence between species is scarce. Learn how to advise owners about safely introducing and keeping members of potential predator and prey species in the same house. PMID- 18409143 TI - Chronic pancreatitis in dogs and cats. AB - Pancreatitis is the most common disorder of the exocrine pancreas in dogs and cats. Clinical diagnosis of chronic pancreatitis is challenging because the disease is usually mild or subclinical and because its clinical signs are often the same as those of complicating or concurrent diseases. Obtaining a detailed history, performing a thorough physical examination, and conducting tests that are sensitive and specific for pancreatitis are crucial in diagnosing chronic pancreatitis. Initial management of an acute episode of chronic pancreatitis largely involves supportive and dietary measures, while long-term management of chronic pancreatitis is based on dietary modification. Management of complications and concurrent diseases is crucial in animals with chronic pancreatitis. PMID- 18409144 TI - The potential role of Akt phosphorylation in human cancers. AB - Akt/protein kinase B (PKB) is a serine/threonine kinase which is implicated in mediating a variety of biological responses including cell growth, proliferation and survival. Akt is activated by phosphorylation on two critical residues, namely threonine 308 (Thr308) and serine 473 (Ser473). Several studies have found Akt2 to be amplified or overexpressed at the mRNA level in various tumor cell lines and in a number of human malignancies such as colon, pancreatic and breast cancers. Nevertheless, activation of Akt isoforms by phosphorylation appears to be more clinically significant than Akt2 amplification or overexpression. Many studies in the past 4-5 years have revealed a prognostic and/or predictive role of Akt phosphorylation in breast, prostate and non-small cell lung cancer. Several publications suggest a role of phosphorylated Akt also in endometrial, pancreatic, gastric, tongue and renal cancer. However, different types of assays were used in these studies. Before assessment of P-Akt can be incorporated into routine clinical practice, all aspects of the assay methodology will have to be standardized. PMID- 18409145 TI - Real-time reverse-transcription PCR to quantify a panel of 19 genes in breast cancer: relationships with sentinel lymph node invasion. AB - At the Centre Oscar Lambret, the anticancer centre of the North of France, sentinel lymph node (SLN) procedures are routinely performed for localized (T0 T1, N0, M0) breast carcinoma without any previous treatment, in order to prevent the deleterious effects of axillary lymph node dissection. The present study was undertaken to assess if the expression in the tumor of a panel of 19 genes would allow to predict histological SLN involvement. We looked at cytokeratin 19 (CK19), mucin-1 (MUC1), mammaglobin (MGB1), cyclin D1 (CCND1), the four members of the HER/ErbB growth factor receptor family (EGFR, HER2-4), insulin-like growth factor-1 receptor (IGF-1R), estradiol receptors (ERalpha, ERbeta), progesterone receptor (PR), vascular endothelial growth factors (VEGF, VEGF-C), urokinase-like plasminogen activator (uPA), matrix metalloproteinases 2 and 9 (MMP2, MMP9), ets related transcription factor ERM, and E-cadherin (CDH1). Their expression was quantified by real-time RT-PCR in 134 breast cancer samples and the relationships with SLN metastases were analyzed. A slight increase (35-40%) in CK19 and HER3 expression was observed in the tumors of patients with SLN metastases compared to those of patients without metastases, even if neither CK19 expression nor HER3 expression allowed to distinguish patients with micrometastases from patients with macrometastases. We conclude that the tumoral expression of biological parameters involved in cell proliferation or playing a critical role in the metastatic process, including tumor invasion and angiogenesis, is not strongly associated with SLN metastases. PMID- 18409146 TI - Relationship between genetic polymorphism of CYP1A1 at codon 462 (Ile462Val) in colorectal cancer. AB - AIMS AND BACKGROUND: The enzyme cytochrome P450 plays an important role in the metabolization and detoxification of various compounds. CYP1A1 is a polymorphic enzyme and some of its alleles have been correlated with an increased risk of developing various types of cancer. The aim of this study was to investigate the incidence of the polymorphism A-->G (Ile462Val, exon 7) in colorectal cancer patients and the correlation of this polymorphism with others risk factors. PATIENTS AND METHODS: 114 Brazilian patients with colorectal cancer were matched by age and sex to 114 healthy individuals. DNA was extracted from peripheral blood and the genotypes of the polymorphisms were assessed by PCR-restriction fragment length polymorphism. RESULTS: In the case group 64 subjects were male, 53 were alcohol users and 68 were smokers. In the control group 61 were male, 67 were alcohol users and 53 smokers. There were 14 subjects with wild-type homozygous A/A, 97 with heterozygous A/G, and 3 with homozygous mutated G/G in the cancer group versus 81 subjects with wild-type homozygous A/A and 33 with heterozygous A/G in the control group. The presence of the G allele (OR 5.14, 95%CI 3.15-10.80) was associated with an increased risk of colorectal cancer (p=0.001). The prevalence of smokers was higher in the cancer group (p=0.047, OR 1.71, 95%CI 1.03-3.11). CONCLUSION: These results suggest a positive association between the A-->G polymorphism and the risk of colorectal cancer. In addition, smoking was also a colorectal cancer risk. We did not find any correlation between this polymorphism and sex, grade of differentiation, stage, or evolution of the disease. PMID- 18409147 TI - CYP1A1 *2B and *4 polymorphisms are associated with lung cancer susceptibility in Mexican patients. AB - BACKGROUND: CYP1A1 is a gene involved in the high aryl hydrocarbon hydroxylase inducible phenotype, which is a genetically-determined variation among individuals that has been associated with lung cancer risk. More specifically, CYP1A1 *2B and *4 polymorphisms have been associated with high susceptibility to lung cancer among cigarette smokers. MATERIALS AND METHODS: DNA was obtained from blood samples and we studied by PCR-RFLP the distribution of CYP1A1 *2B (n=248) and *4 (n=222) polymorphisms in healthy controls and 222 lung cancer patients from a Mexican population. RESULTS: Comparisons between groups showed an increased risk for lung cancer patients of *2B/*2B (18%; OR 7.6; 95% CI 3.0-19.2) and *4/ *4 genotypes (15%; OR 11.45; 95% CI 2.19-59.85) compared to the control group (1% for *2B/ *2B and 4.4% for *4/ *4). A significant association between lung cancer and homozygous *2B/ *2B passive smokers and *4/*4 ever (cigarettes) and passive smokers was also observed (p<0.05). Multivariate analysis revealed an increased risk for the *2B/*2B genotype (OR 6.83), as well as for *4/*4 (OR 28.8). CONCLUSION: The results of the study indicate a significant association between *2B/*2B and *4/*4 genotypes and the risk of developing lung cancer among Mexicans. PMID- 18409148 TI - Antiphospholipid antibodies and acute-phase response in non-metastatic colorectal cancer patients. AB - AIM: To investigate the plasma levels and prevalence of the most common antiphospholipid antibodies, as well as their relationships with several plasma markers of inflammation, in order to characterize some aspects of cancer thrombophilia. MATERIALS AND METHODS: Eighty-three cancer patients with non metastatic colorectal solid tumors and 94 control subjects were tested for the presence of IgG/IgM/IgA anti-cardiolipin and anti-Beta2-glycoprotein I antibodies and of several acute-phase reactants, i.e., fibrinogen, factor VIII:C and C4b binding protein. RESULTS: In cancer patients the plasma levels of the acute-phase reactants and the IgA/IgG anti-cardiolipin and IgA anti-Beta2- glycoprotein I antibodies were significantly higher; the acute-phase reactants were significantly correlated with anti-cardiolipin antibodies; the prevalence of antiphospholipid antibodies was not significantly higher. CONCLUSIONS: In patients with non-metastatic colorectal cancer the acute-phase response is associated with antiphospholipid generation. This could represent a further pathogenetic mechanism for the short-term post-surgery thrombotic complications of patients with colorectal cancer. PMID- 18409149 TI - Coagulation factor levels in non-metastatic colorectal cancer patients. AB - There is evidence that high plasma levels of factor (F) VIII, FIX, FXI and fibrinogen are independent risk factors for venous thromboembolism. AIM: To determine the plasma concentrations of several coagulation factors and C4b binding protein (C4BP) in a group of patients with non-metastatic colorectal cancer in order to investigate some aspects of cancer-acquired thrombophilia. METHODS: Plasma fibrinogen, FII, FV, FVII, FVIII, FIX, FX, FXI and FXII activity levels and C4BP concentrations were determined in 73 patients with non-metastatic colorectal cancer (48 colon and 25 rectum) and in 67 matched control subjects. No one in either group had had previous thrombotic events. RESULTS: Mean plasma concentrations of fibrinogen (functional and antigen), FVIII, FIX, FV and C4BP were significantly higher in colorectal cancer patients than in control subjects, while FVII and FXII levels were significantly decreased. Several correlations were found between the increased coagulation factors and C4BP concentrations, while FVII was highly correlated with FXII. CONCLUSIONS: In colorectal cancer patients high plasma fibrinogen, FVIII and FIX levels might represent further risk factors for venous thrombotic complications in the immediate post-surgery period, while decreased FVII and FXII concentrations may be an index of intravascular coagulation activation, still in a subclinical phase. PMID- 18409150 TI - Assessment of the biological variation of plasma tissue inhibitor of metalloproteinases-1. AB - BACKGROUND: Tissue inhibitor of metalloproteinases-1 (TIMP-1) measurements in plasma may be useful for the early detection and prognosis of colorectal cancer (CRC). Data on analytical performance and normal intra- and interindividual biological variation are required in order to interpret the utility of TIMP-1 in CRC. The aim of this study was to establish the biological and analytical variation of plasma TIMP-1 in volunteers. MATERIAL AND METHODS: Three separate studies were undertaken. 1: Plasma was collected from 23 volunteers 6 times within a 3-week period, first in September 2004 (round [R] 1), then repeated in May 2005 (R2) and May 2006 (R3) in the same group of individuals. TIMP-1 levels were determined by the MAC15 ELISA assay and with the Abbott ARCHITECT i2000 Immunoanalyzer. 2: Circadian variation was evaluated in plasma collected 7 times within a 24-hour period (n=16). 3: Effects of physical exercise were evaluated in plasma collected before and after bicycling (n=14). In studies 2 and 3 TIMP-1 levels were determined with the MAC15 ELISA assay only. RESULTS: A significant correlation between TIMP-1 MAC15 and ARCHITECT i2000 was shown (rs=0.78, p<0.002), with consistently higher levels being detected by the ARCHITECT i2000. Median levels of TIMP-1 (ARCHITECT) at 8 a.m. in each round were 74.9 ng/mL (range 65.7-89.9) (R1), 87.3 ng/mL (range 72.7-127.9) (R2), and 81.9 ng/mL (range 66.8-113.6) (R3). The within-subject variation was 10.7%, the variation between rounds was 7.4%, and the intraclass correlation was 46.2%. Comparison between the 3 rounds and time of collection showed that TIMP-1 values decreased by 11% after storage for more than 16 months (p=0.0002). A systematic circadian variation in plasma TIMP-1 levels was not observed (p=0.17). No significant variation of plasma TIMP-1 was found in relation to physical exercise (p=0.92 [global test]). CONCLUSION: Levels of plasma TIMP-1 in volunteers show limited circadian, day-to day, week-to-week and season-to-season variation. In addition, physical exercise has no impact on plasma TIMP-1 levels. Possible storage-dependent decreases in plasma TIMP-1 levels warrant further investigation. PMID- 18409151 TI - Identification of alphaB-crystallin, a biomarker of renal cell carcinoma by SELDI TOF MS. AB - Spectrometric-based surface-enhanced laser desorption/ionization ProteinChip (SELDI-TOF) facilitates rapid and easy analysis of protein mixtures and is often exploited to define potential diagnostic markers from sera. However, SELDI- TOF is a relatively insensitive technique and unable to detect circulating proteins at low levels even if they are differentially expressed in cancer patients. Therefore, we applied this technology to study tissues from renal cell carcinomas (RCC) in comparison to healthy controls. We found that different biomarkers are identified from tissues than those previously identified in serum, and that serum markers are often not produced by the tumors themselves at detectable levels, reflecting the nonspecific nature of many circulating biomarkers. We detected and characterized aB-crystallin as an overexpressed protein in RCC tissues and showed differential expression by immunohistochemistry. We conclude that SELDI-TOF is more useful for the identification of biomarkers that are synthesized by diseased tissues than for the identification of serum biomarkers and identifies a separate set of markers. We suggest that SELDI-TOF should be used to screen human cancer tissues to identify potential tissue-specific proteins and simpler and more sensitive techniques can then be applied to determine their validity as biomarkers in biological fluids. PMID- 18409152 TI - Circulating cytokeratin 19 fragments in patients with benign nodules and carcinomas of the thyroid gland. AB - Cytokeratin 19 (CK19) is an acidic protein of 40 kDa that is part of the cytoskeleton of epithelial cells and is highly expressed by differentiated thyroid carcinomas, mainly of the papillary subtype. The soluble fragments of CK19 (Cyfra 21.1) can be measured by immunometric assays employing specific monoclonal antibodies. The present study was planned to assess the serum expression of Cyfra 21.1 in patients with benign thyroid nodules and thyroid malignancies. We enrolled 135 patients with histologically proven benign thyroid nodules (n=79) and thyroid carcinomas (n=56). No differences were found in serum Cyfra 21.1 levels between patients with benign nodules and patients with carcinomas. When thyroid malignancies were subdivided according to tumor histology, serum Cyfra 21.1 increased significantly from classical differentiated thyroid carcinomas (papillary or follicular) to less differentiated or undifferentiated carcinomas (poorly differentiated or anaplastic). CK19 release into the bloodstream is strongly related to the apoptotic pathway, and particularly to hyperproliferation-related apoptosis. These pathways characterized anaplastic and poorly differentiated thyroid carcinoma but not classical forms of differentiated thyroid carcinoma. Consequently, Cyfra 21.1 may be regarded as a circulating marker of poorly differentiated and anaplastic thyroid carcinoma. Additionally, a role of Cyfra 21.1 as a dedifferentiation marker in patients with classical differentiated thyroid carcinomas may be postulated and should be explored by further focused studies. PMID- 18409153 TI - Reduced clinicopathological influence of hormone-dependence on breast carcinomas in women older than 70 years. AB - In order to evaluate the influence of hormone dependence on the features of infiltrating ductal carcinoma of the breast we have assayed the cytosolic levels of estrogen receptor (ER), progesterone receptor (PR), pS2 and cathepsin D in 53 women aged over 70 years and in 95 women aged between 55 and 70 years. Tumor size, axillary involvement, distant metastasis, histological grade, ploidy and S phase were taken into account. Carcinomas of women aged over 70 did not show higher concentrations or higher positive results for ER and PR than those of women in the 55-70-year age group. In older patients, negativity for ER was associated only with higher S-phase fraction, while negativity for PR was not associated with any of the parameters analyzed. In the younger subgroup, negativity for ER was associated with larger tumor size, higher S-phase fraction, lymph node involvement, histological grade 3 and lower pS2 values. Negativity for PR was associated with the same parameters, as well as with a higher frequency of recurrence. Our results suggest a reduced influence of hormone dependence on the clinicopathological features of breast carcinomas in patients older than 70 years compared with women aged between 55 and 70 years. PMID- 18409156 TI - In-line solid-phase extraction preconcentration in capillary electrophoresis tandem mass spectrometry for the multiresidue detection of quinolones in meat by pressurized liquid extraction. AB - We have developed and validated a CE-MS/MS method using an in-line SPE device (analyte concentrator, AC) to determine eight quinolones of veterinary use whose maximum residue levels in animal edible tissues are established by the EU Council Regulation 2377/90, i.e., danofloxacin, sarafloxacin, ciprofloxacin, marbofloxacin, enrofloxacin, difloxacin, oxolinic acid, and flumequine. Different parameters affecting the AC performance, such as its design (in this case frit free), the kind of sorbent (Oasis MCX), sample pH, volume, and composition of the elution plug and injection time were studied. The method was validated using standard solutions obtaining LODs between 17 and 59 ng/L. Finally, a pressurized liquid extraction (PLE) method was developed to determine these antibiotics in chicken muscle samples. The whole analytical method was validated in terms of linearity (r2 >or= 0.992), recoveries (63-112%), repeatability and intermediate precision (RSD 1 ppm) and allowed high throughput. In contrast, the combination of liquid-liquid extraction (LLE) and electrokinetic injection (EK) was time consuming but did allow detection at the ppb level. Both approaches were fully validated according to ICH guidelines and SFSTP protocols for two pharmaceutical compounds (ecstasy and methadone (MTD)). Deuterated internal standards (IS) in the analytical procedures were used and good quantitative performance was obtained in terms of trueness and precision (repeatability and intermediate precision) since accuracy profiles were within the acceptance limits (30% for biological assay). Methods were linear over the concentration range of 0.50-175 ng/mL and 0.25-5 microg/mL for LLE-EK and PP-HD procedures, respectively. The LLE EK methodology was finally successfully applied to quantitation of ecstasy and MTD in real cases obtained from toxicology. PMID- 18409163 TI - Separation of cytokinin isomers with a partial filling-micellar electrokinetic chromatography-mass spectrometry approach. AB - A new method based on partial filling-MEKC (PF-MEKC) directly coupled to ESI-MS was developed for the simultaneous separation and determination of 13 structurally similar cytokinins, including various geometric and positional isomers of cytokinins. On the basis of the resolution of the neighboring isomer peaks, different parameters (i.e., pH and concentration of buffer, surfactant concentrations, length of the injected micellar plug, organic modifier, and applied separation voltage) were optimized to achieve a satisfactory PF-MEKC separation. Under optimum conditions, the separation of 13 cytokinin standards was accomplished within 25 min. MS/MS with multiple reaction monitoring detection was carried out to obtain sufficient selectivity. PF-MEKC-MS/MS allowed for the direct identification and confirmation of the cytokinins present in banana (Musa spp.) pulp sample after extraction and purification. Finally, trans-zeatin riboside (ZR) and trans-zeatin (Z) were unambiguously identified in banana pulp. It is anticipated that the current PF-MEKC-MS method can be applied to analyze cytokinins in a wide range of biological samples. PMID- 18409164 TI - Development of a quantitative, validated capillary electrophoresis-time of flight mass spectrometry method with integrated high-confidence analyte identification for metabolomics. AB - A CE-MS method was developed and validated for the quantitative analysis of negatively charged metabolites by making use of the high mass accuracy and the quantitation capabilities of a TOF mass analyzer in combination with automated feature extraction and database search. Metabolites of the central carbon metabolism were quantified with an LOD and lower LOQ (LLOQ) of 0.2-2 and 1-4 microM, respectively. The method was used to elucidate metabolic changes in the Escherichia coli deletion mutant PntAB-UdhA that lacks nicotinamide nucleotide transhydrogenase function, under both stationary and exponential growth conditions. The reproducibility of metabolite extraction and CE-TOF-MS analysis ranged from 3.7 to 22.7 and 7.9 to 22.6%, respectively, while the biological variance was 3.4-31.3%. We observed significant differences in metabolite abundance, particularly in the citrate cycle, between wild-type and mutant E. coli. Overall, more than 600 features were found by automated feature detection, which resulted in approximately 150 high-confidence metabolite identifications. Concomitant analyses with two different GC-MS methods allowed not only crossvalidation of the quantitative results obtained by the various methods, but also led to a more comprehensive coverage of the E. coli metabolome. PMID- 18409165 TI - Electrical field-assisted solid-phase extraction coupled on-line to capillary electrophoresis-mass spectrometry. AB - A substantial demand currently exists for analytical methods affording the determination of very low concentrations of analytes in complex matrices, such as those of environmental and biological samples, as simply as possible. However, the pretreatment of complex samples, which is unavoidable prior to CE-MS analysis, is usually complicated and time-consuming. In this work, we used voltage-assisted SPE for the first time as an alternative to conventional treatments for preconcentrating and purifying analytes. To this end, we used a simple flow system coupled on-line to CE-MS equipment. The system is quite robust and provides reproducible peak areas (the precision ranges from 2.5 to 3.8%). Also, it provides increased sensitivity affording the determination of trace amounts (nanogram per liter levels) of analytes in only a few milliliters of sample. The proposed system was applied to the determination of members of two compound families (viz. tetracyclines and amines). PMID- 18409166 TI - Serum protein profiling in mice: identification of Factor XIIIa as a potential biomarker for muscular dystrophy. AB - Protein profiling in blood serum by fractionation and MS analysis has been applied in mice to assess its applicability as a fast, economical alternative to current DNA and RNA analyses for diagnosis of neuromuscular disorders. Mass spectra of peptides and proteins were generated using serum from dystrophin deficient mdx and control mice by WCX ClinProt bead fractionation, followed by MALDI-MS. Double cross-validatory linear discriminant and logistic regression data analysis methods were compared with a new Bayesian logistic regression method. These were evaluated on their ability to discriminate between healthy and dystrophic samples, and to identify the discriminatory peaks in the mass spectra. All three approaches classified the spectra with comparable misclassification rates (between 18.4 and 20.6%), with much overlap between the differential peaks identified between the methods. The differential peak pattern from the Bayesian method was sparser and easier to interpret than from the other two methods, without compromising classifying strength. One of the two main differentiating peaks at m/z 3908 was identified as an N-terminal peptide of coagulation Factor XIIIa, previously identified in human serum. This work underlines the translational aspect of serum protein profiling in mice and supports a further study with serum from patients with neuromuscular disorders. PMID- 18409167 TI - Application of 16O/18O reverse proteolytic labeling to determine the effect of biofilm culture on the cell envelope proteome of Porphyromonas gingivalis W50. AB - Porphyromonas gingivalis is an oral pathogen linked to chronic periodontitis. The bacterium exists as part of a polymicrobial biofilm accreted onto the tooth surface. An understanding of the changes to the proteome especially of the cell envelope of biofilm cells compared with planktonic cells could provide valuable insight into the molecular processes of biofilm formation. To establish which proteins changed in abundance between the planktonic and biofilm growth states, the cell envelope fractions of two biological replicates of P. gingivalis cultivated in a chemostat were analysed. Proteins were separated by 1-D SDS-PAGE, in-gel digested with trypsin in the presence of H216O or H218O and identified and quantified by LC-MALDI TOF/TOF-MS. Using a reverse labeling strategy we identified and quantified the changes in abundance of 81 P. gingivalis cell envelope proteins. No form of bias between the labels was observed. Twenty four proteins increased in abundance and 18 decreased in abundance in the biofilm state. A group of cell-surface located C-Terminal Domain family proteins including RgpA, HagA, CPG70 and PG99 increased in abundance in the biofilm cells. Other proteins that exhibited significant changes in abundance included transport related proteins (HmuY and IhtB), metabolic enzymes (FrdAB) and immunogenic proteins. PMID- 18409168 TI - Proteomic investigation of the adaptation of Lactococcus lactis to the mouse digestive tract. AB - Lactic acid bacteria are used on an industrial scale for the manufacturing of dairy products. It is now intended to develop novel applications of lactic acid bacteria that could be used as living vehicles for the targeting of antigens or therapeutics to the digestive mucosa. The aim of this study was to analyze the adaptations of Lactococcus lactis, a model lactic acid bacteria to the digestive tract and to identify functions required for colonization of the intestine. For this purpose, we combined gnotobiology with proteomics: axenic mice were colonized with a dairy L. lactis strain and the bacterial proteome was examined by 2-DE. As compared to cultures in broth, the proteome profile of bacteria grown in the intestine indicates the activation of metabolic pathways involved in various carbon sources assimilation and suggests the adoption of a mixed acids fermentative metabolism. We identified the product of the ywcC gene as essential for the colonization of the digestive tract and demonstrated that the corresponding gene product (YwcC) possesses a phosphogluconolactonase activity, suggesting an important role of the pentose phosphate pathway for the development of L. lactis in the digestive environment. PMID- 18409172 TI - Is alpha-lipoic acid a scavenger of reactive oxygen species in vivo? Evidence for its initiation of stress signaling pathways that promote endogenous antioxidant capacity. AB - The chemical reduction and oxidation (redox) properties of alpha-lipoic acid (LA) suggest that it may have potent antioxidant potential. A significant number of studies now show that LA and its reduced form, dihydrolipoic acid (DHLA), directly scavenge reactive oxygen species (ROS) and reactive nitrogen species (RNS) species and protect cells against a host of insults where oxidative stress is part of the underlying etiology. However, owing to its limited and transient accumulation in tissues following oral intake, the efficacy of nonprotein-bound LA to function as a physiological antioxidant has been questioned. Herein, we review the evidence that the micronutrient functions of LA may be more as an effector of important cellular stress response pathways that ultimately influence endogenous cellular antioxidant levels and reduce proinflammatory mechanisms. This would promote a sustained improvement in cellular resistance to pathologies where oxidative stress is involved, which would not be forthcoming if LA solely acted as a transient ROS scavenger. PMID- 18409173 TI - Estrogen is a modulator of vascular inflammation. AB - Vascular inflammation underlies the pathogenesis of atherosclerosis. Atherosclerotic changes in the vasculature lead to conditions such as coronary artery disease and stroke, which are the major causes of morbidity and mortality worldwide. Epidemiological studies in premenopausal women suggest a beneficial role for estrogen in preventing vascular inflammation and consequent atherosclerosis. However, the benefits of estrogen areabsent or even reversed in older postmenopausal subjects. The modulation of inflammation by estrogen under different conditions might explain this discrepancy. Estrogen exerts its antiinflammatory effects on the vasculature through different mechanisms such as direct antioxidant effect, generation of nitric oxide, prevention of apoptosis in vascular cells and suppression of cytokines and the renin-angiotensin system. On the other hand, estrogen also elicits proinflammatory changes under certain conditions, which are less completely understood. Some of the mechanisms underlying a possible proinflammatory role for estrogen include increased expression of the proinflammatory receptor for advanced glycation end products, increased tyrosine nitration of cellular proteins, and generation of reactive oxygen species through an uncoupled eNOS. In this review, we have presented evidence for both antiinflammatory and proinflammatory pathways modulated by estrogen and how interactions among such pathways might determine the effects of estrogen on the vascular system. PMID- 18409175 TI - On the way to selective PARP-2 inhibitors. Design, synthesis, and preliminary evaluation of a series of isoquinolinone derivatives. AB - PARP-1 and PARP-2 are members of the family of poly(ADP-ribose)polymerases, which are involved in the maintenance of genomic integrity under conditions of genotoxic stimuli. The different roles of the two isoforms under pathophysiological conditions have not yet been fully clarified, and this is partially due to the lack of selective inhibitors. We report herein the synthesis and preliminary pharmacological evaluation of a large series of isoquinolinone derivatives as PARP-1/PARP-2 inhibitors. Among them, we identified the 5 benzoyloxyisoquinolin-1(2 H)-one derivative as the most selective PARP-2 inhibitor reported so far, with a PARP-2/PARP-1 selectivity index greater than 60. PMID- 18409176 TI - Effects of peripheral chloro substitution on the photophysical properties and in vitro photodynamic activities of galactose-conjugated silicon(IV) phthalocyanines. AB - A series of silicon(IV) phthalocyanines with two axial isopropylidene-protected galactose moieties and one, two, or eight chloro group(s) on the periphery of the macrocycle have been synthesised and spectroscopically characterised. The photophysical properties and in vitro photodynamic activities of these compounds have been studied and compared with those of the nonchlorinated analogue. All the compounds, with the exception of the octachlorinated counterpart which has a limited solubility, are essentially nonaggregated in N,N-dimethylformamide. The fluorescence quantum yield decreases and the singlet oxygen quantum yield increases as the number of chloro substituent increases, which is in accord with the heavy-atom effect. The non-, mono-, and dichlorinated phthalocyanines formulated with Cremophor EL are all photodynamically active against HT29 human colon adenocarcinoma and HepG2 human hepatocarcinoma cells with IC(50) values ranging from 0.03 to 1.05 microM. The photocytotoxicity as well as the efficiency to generate intracellular reactive oxygen species decrease along this series because of the increase in aggregation tendency upon chloro substitution. The nonchlorinated analogue exhibits the highest potency and can target the lysosomes of HT29 cells, whereas the monochlorinated counterpart is not localised in the lysosomes. PMID- 18409177 TI - Crystal structures of a second g protein-coupled receptor: triumphs and implications. PMID- 18409178 TI - A new glycorotaxane molecular machine based on an anilinium and a triazolium station. PMID- 18409179 TI - Mutations in the chromatin-associated protein ATRX. AB - ATRX belongs to the SNF2 family of proteins, many of which have been demonstrated to have chromatin remodeling activity. Constitution mutations in the X-encoded gene give rise to alpha thalassemia mental retardation (ATR-X) syndrome and a variety of related conditions that are often associated with profound developmental delay, facial dysmorphism, genital abnormalities, and alpha thalassemia. Acquired mutations in ATRX are observed in the preleukemic condition alpha thalassemia myelodysplastic syndrome (ATMDS). Mutations in ATRX have been shown to perturb gene expression and DNA methylation. This is a comprehensive report of 127 mutations including 32 reported here for the first time. Missense mutations are shown to cluster in the two main functional domains. The truncating mutations appear to be "rescued" to some degree and so it appears likely that most if not all constitutional ATRX mutations are hypomorphs. PMID- 18409180 TI - Rhombencephalosynapsis: prenatal imaging and autopsy findings. AB - OBJECTIVES: Rhombencephalosynapsis is a rare, but increasingly recognized, brain malformation characterized by congenital fusion of the cerebellar hemispheres and absence of the vermis. Rhombencephalosynapsis is associated with significant developmental delay, seizures and involuntary head movements. We report four cases, with correlation of prenatal and postnatal imaging and autopsy findings. METHODS: Over a 2-year period, four cases of rhombencephalosynapsis were diagnosed in the perinatal period, three in one center and one in another center. The clinical cases were reviewed, and correlation was made between the prenatal and postnatal imaging and autopsy findings where available. RESULTS: All cases presented initially with ventriculomegaly on prenatal ultrasound examination. Subsequent magnetic resonance imaging (MRI) established the diagnosis in two cases and postnatal MRI established the diagnosis in a further two cases. Autopsy was available and confirmed the diagnosis in two cases. In one case the pregnancy was terminated, two infants died in the neonatal period and one died in infancy. CONCLUSIONS: The cases in this perinatal series of rhombencephalosynapsis showed a very poor prognosis. The presence of ventriculomegaly on prenatal ultrasound imaging should alert the physician to consider rhombencephalosynapsis in the differential diagnosis. MRI appears to be the imaging modality of choice in establishing the diagnosis. PMID- 18409181 TI - Color M-mode propagation velocity, but not its ratio to early diastolic inflow velocity, changes throughout gestation in normal human fetuses. AB - OBJECTIVES: Color M-mode propagation velocity (Vp) is a measure of diastolic function in adults and, when combined with early diastolic inflow velocity (E), the ratio E/Vp reflects ventricular filling pressure. Early detection of diastolic compromise may benefit fetal patients at risk for developing heart failure. The objectives of this study were to measure values for Vp and inflow peak E in a group of normal fetuses, to analyze age-dependent alterations in these measurements, and to evaluate the interobserver and intraobserver variability of the measurements. METHODS: Thirty-two normal fetuses at between 20 and 35 weeks' gestation underwent echocardiography. Color M-mode Vp was measured from the four-chamber view for the right (RV) and left (LV) ventricles, and mitral and tricuspid inflow velocities were determined by pulsed-wave Doppler ultrasound. The values obtained were compared with previously reported findings in adults. RESULTS: Adequate tracings were obtainable in 23 patients for the RV and 29 for the LV. Mean Vp values for the RV (15.3 +/- 3.2 cm/s) and LV (20.8 +/- 5.6 cm/s) were lower than normal adult values, and Vp values were significantly lower for the RV than the LV (P < 0.001). Applying Bazett's heart rate correction, values for RV (23.4 +/- 4.8 cm/s) and LV (31.9 +/- 8.7 cm/s) remained lower than normal adult values. There was a linear correlation of Vp with gestational age for the RV (R = 0.69, P < 0.001), and the ratio of E/Vp corrected for heart rate for the RV (1.51 +/- 0.26) remained constant throughout gestation. Interobserver bias was high but intraobserver bias low, at 19 and 1.1%, respectively. CONCLUSIONS: Vp is lower in fetal than in adult life. Vp for the RV changes in a manner indicative of improving diastolic function throughout normal gestation, providing insight into the alterations in diastolic function with gestation that contribute to increases in cardiac output. The use of Vp to assess diastolic function disturbance in fetuses is feasible, but high interobserver variability is problematic. PMID- 18409182 TI - Psychometric properties of the Retrospective Self Report of Inhibition (RSRI) in a representative German sample. AB - The present study examined the internal consistency, factorial structure, and construct validity of the German version of the Retrospective Self Report of Inhibition (RSRI), a questionnaire measure of behavioral inhibition. The research was based on data from a German prospective-longitudinal community study of 3021 adolescents and young adults (aged 14-24 years at baseline). Diagnostic assessment was based on the DSM-IV/M-CIDI and general psychopathological distress was assessed with SCL-90-R. Results of confirmatory factor analysis indicated adequate fit of the two-factor model, suggested by the authors of the original version. Indices of internal consistency of the RSRI and its subscales 'social/school' and 'fear/illness' were shown to be sufficient for the total sample and even higher in subgroups of subjects with certain DSM-IV diagnoses. Associations with variables such as mental distress, parental psychopathology, and DSM-IV disorders were in line with theoretical assumptions and confirm different aspects of the validity (convergent, concurrent, predictive) of the instrument. The psychometric properties of the German RSRI were found to be comparable to those of the English version. The applicability of this questionnaire in German-speaking countries is therefore recommended for adolescents and young adults. PMID- 18409183 TI - Defecation proctography and translabial ultrasound in the investigation of defecatory disorders. AB - OBJECTIVES: Defecation proctography is the standard method used in the investigation of obstructed defecation. Translabial ultrasound has recently been shown to demonstrate rectocele, enterocele and rectal intussusception. We performed a comparative clinical study to determine agreement between the two methods. METHODS: Thirty-seven women scheduled to undergo defecation proctography for obstructed defecation were recruited. Using both proctography and translabial ultrasound, we determined the anorectal angle, presence of a rectocele and rectocele depth, rectal intussusception and prolapse. Measurements were obtained by operators blinded to all other data. All patients rated discomfort on a scale of 0-10. RESULTS: Six women did not attend defecation proctography, leaving 31 cases for comparison. The mean age was 53 years. Patients rated discomfort at a median of 1 (range 0-10) for ultrasound and 7 (range 0-10) for defecation proctography (P < 0.001). Defecation proctography suggested rectocele and rectal intussusception/prolapse more frequently than did ultrasound. While the positive predictive value of ultrasound (considering defecation proctography to be the definitive test) was 0.82 for rectocele and 0.88 for intussusception/prolapse, negative predictive values were only 0.43 and 0.27, respectively. Cohen's kappa values were 0.26 and 0.09, respectively. There was poor agreement between ultrasound and defecation proctography measurements of anorectal angle and rectocele depth. CONCLUSIONS: Translabial ultrasound can be used in the initial investigation of defecatory disorders. It is better tolerated than defecation proctography and also yields information on the lower urinary tract, pelvic organ prolapse and levator ani. Agreement between ultrasound and defecation proctography in the measurement of quantitative parameters was poor, but when intussusception or rectocele was diagnosed on ultrasound these results were highly predictive of findings on defecation proctography. PMID- 18409184 TI - Back injuries among union carpenters in Washington State, 1989-2003. AB - BACKGROUND: There is limited information on occupational back pain specific to carpenters despite their known exposures to recognized occupational risk factors and limited opportunities for modified work due to the predominantly heavy nature of their work. METHODS: By combining union records with worker's compensation claims, we describe work-related back injuries, including associated medical diagnoses, among a well-defined cohort of union carpenters between 1989 and 2003. High risk subgroups were explored based on age, gender, union tenure, and predominant type of work. Paid lost time claims were contrasted to less serious events, and injuries sustained from overexertion activities were contrasted with those sustained through more acute trauma. RESULTS: Back injuries occurred at an overall rate of 6.2/200,000 hours worked. Most injuries were coded in the compensation records as sprains, but there was little agreement between these nature of injury codes and ICD9 diagnosis codes. Injury rates declined most significantly over time for injuries secondary to overexertion. In multivariate analyses, we observed similar patterns of risk for the types of claims evaluated despite disparate mechanisms and severity. Those who worked predominantly in residential carpentry or drywall installation were consistently at greatest risk. CONCLUSIONS: Overexertion injuries from manual materials handling activities are responsible for the largest burden of back injuries among these carpenters, but a growing proportion of injuries result from acute traumatic events. Interventions are called for which specifically address risk among residential carpenters and drywall installers. These data provide additional evidence that Bureau of Labor Statistics data underestimate work-related injuries. PMID- 18409185 TI - Reassessment of occupational exposure limits. AB - BACKGROUND: Although the Netherlands currently has its own procedure for evaluating chemical compounds and setting occupational exposure limits (OELs), most of these limits were originally adopted in the 1970s from threshold limit values (TLVs) set by the American Conference of Governmental Industrial Hygienists (ACGIH). However, beginning in the late 1980s, criticism about non scientific considerations being used to set TLV's suggested that TLVs might not offer sufficient health protection to workers. This situation prompted the Dutch Ministry of Social Affairs and Employment to request that the Health Council of the Netherlands reassess the health protection of MAC values that were contained in the 1994 Dutch MAC list. METHODS: Criteria documents were prepared for 161 compounds. They were evaluated by a committee of the Health Council of the Netherlands consisting of international experts who reassessed the toxicological hazards of these substances and recommended, whenever possible, health-based OELs. The results of the reassessment by the Health Council were compared with the MAC values of the 1994 Dutch MAC list, ACGIH TLVs, and existing German OELs. RESULTS: The toxicological database met the committee's criteria for a health based OEL for only about 40% of the compounds. CONCLUSIONS: Many older MAC values were either too high or not scientifically supported and therefore not health based. PMID- 18409186 TI - The interobserver variability of digital rectal examination in a large randomized trial for the screening of prostate cancer. AB - BACKGROUND: To analyze to what extent the percentage of suspicious digital rectal examination (DRE) findings vary between examiners and to what extent the percentage of prostate cancers (PCs) detected in men with these suspicious findings varies between examiners. METHODS: In the first screening round of the European Randomized study of Screening for PC (ERSPC) Rotterdam, 7,280 men underwent a PSA-determination and DRE of whom 2,102 underwent prostate biopsy (biopsy indication PSA > or = 4.0 ng/ml and/or suspicious DRE and/or TRUS). Descriptive statistics of DRE-outcome per PSA-range were used to determine the observer variability of six examiners. Because this analysis did not correct properly for other predictors of a suspicious DRE (PSA-level, biopsy indication, TRUS-outcome, prostate volume and age), a logistic regression analysis controlling for these explanatory variables was performed as well. RESULTS: In 2,102 men biopsied, 443 PCs were detected (PPV = 21%). For all PSA levels the percentage suspicious DRE varied between examiners from 4% to 28% and percentage PC detected in men with a suspicious DRE varied from 18% to 36%. Logistic regression analysis showed that three of six examiners considered DRE significantly more often abnormal than others (ORs 3.48, 2.80, 2.47, P < 0.001). For all examiners the odds to have PC was statistically significantly higher in case of a suspicious DRE (ORs 2.21-5.96, P < 0.05). This increased chance to find PC was not significantly observer-dependent. CONCLUSIONS: Three of six examiners considered DRE significantly more often suspicious than the others. However, under equal circumstances a suspicious DRE executed by each examiner increased the chance of the presence of PC similarly. PMID- 18409187 TI - Kinematics of turning maneuvers in the southern flying squirrel, Glaucomys volans. AB - The ability to change direction is essential to any animal that moves around in a complex, 3D environment. In this study we present the first 3D description of body positions during gliding turns in a mammalian gliding specialist, the southern flying squirrel, Glaucomys volans. In addition, we used these kinematic data to estimate the aerodynamic forces generated by the animals and rotational velocities and accelerations of the body while turning. These results were compared with similar measurements of flying squirrels during straight glides. The two individuals used in this study differed significantly in limb position asymmetries between the two sides of their bodies and also were significantly different in measures of turning performance. The individual with better performance used limb positions consistent with a primarily lift-based turning mechanism (banked turn), whereas the individual with poorer performance used limb positions consistent with a primarily drag-based turning mechanism (crabbed turn). Both individuals employed limb movements continuously through the gliding turn, but these movements did not have any consistent relationships with body rotations or lateral acceleration. As compared with straight glides, squirrels used significantly higher angles of attack and had lower lift-to-drag ratios, but did not differ in glide angle. Contrary to the typical view of maneuvering during gliding as a simple, static form of locomotion, the results presented here indicate that mammalian gliding is a complex behavior comprising the interplay of many components of limb position and wing shape that affect the balance of forces that control the turn. PMID- 18409188 TI - A DNA resequencing array for pathogenic mutation detection in hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HCM) is a heterogeneous autosomal dominant cardiac disorder with a prevalence of 1 in 500. Over 450 different pathogenic mutations in at least 16 genes have been identified so far. The large allelic and genetic heterogeneity of HCM requires high-throughput, rapid, and affordable mutation detection technologies to efficiently integrate molecular screening into clinical practice. We developed a custom DNA resequencing array that contains both strands of all coding exons (160), splice-site junctions, and 5'UTR regions of 12 genes that have been clearly implicated in HCM (MYH7, MYBPC3, TNNT2, TPM1, TNNI3, MYL3, MYL2, CSRP3, PLN, ACTC, TNNC1, and PRKAG2). We analyzed a first series of 38 unrelated patients with HCM (17 familial, 21 sporadic). A total of 953,306 bp across the 38 patients were sequenced with a mean nucleotide call rate of 96.92% (range: 93-99.9%). Pathogenic mutations (single nucleotide substitutions) in MYH7, MYBPC3, TNNI3, and MYL3 (six known and six novel) were identified in 60% (10/17) of familial HCM and 10% of sporadic cases (2/21). The high-throughput HCM resequencing array is the most rapid and cost-effective tool for molecular testing of HCM to date; it thus has considerable potential in diagnostic and predictive testing, and prognostic stratification. PMID- 18409189 TI - Epidermal growth factor receptor (EGFR) expression in prostatic adenocarcinoma after hormonal therapy: a fluorescence in situ hybridization and immunohistochemical analysis. AB - BACKGROUND: The progression of normal prostatic epithelium to androgen-dependent cancer and, eventually, hormone-refractory prostate cancer is a complex process involving many different growth regulatory signals. Activation of epidermal growth factor receptor (EGFR) has been implicated in prostate cancer cell growth. METHODS: This study was undertaken to investigate both amplification of EGFR gene by fluorescence in situ hybridization (FISH) and over-expression of EGFR by immunohistochemical staining in prostate tissue from 71 patients treated by hormonal therapy. RESULTS: EGFR gene amplification was present in 1 of 71 tumors, and polysomy of chromosome 7 was present in 24 of 71 tumors. Immunohistochemically, EGFR expression was demonstrable in 57 of 71 tumors. Membranous immunostaining for EGFR was observed in >75% of tumor cells in 11% of cases, in 51-75% of tumor cells in 20% of cases, in 26-50% of tumor cells in 21% of cases, in 11-25% of tumor cells in 21% of cases, and in 1-10% of tumor cells in 7% of cases. No immunostaining for EGFR was seen in 20% of cases. There was no correlation between EGFR protein expression and gene amplification. There was also no correlation between EGFR expression and clinicopathological characteristics or clinical outcome. CONCLUSIONS: We found that EGFR gene expression was detectable in 35% of this large series of hormone-treated prostate cancer, and that EGFR protein is frequently expressed in tissue from these patients. EGFR over-expression may serve as a reasonable target for therapeutic intervention in this otherwise difficult to treat subset of prostate cancer. PMID- 18409191 TI - The infevers autoinflammatory mutation online registry: update with new genes and functions. AB - Infevers (Internet Fevers; http://fmf.igh.cnrs.fr/ISSAID/infevers), a website dedicated to mutations responsible for hereditary autoinflammatory diseases, was created in 2002 and has continued to evolve. This new version includes eight genes; six were already present: MEFV, MVK, TNFRSF1A, NLRP3, NOD2, PSTPIP1, and two are new, LPIN2 and NLRP7. Currently, Infevers contains over 540 sequence variants. Several new database functions were recently instituted. The website now accepts confidential data and complex alleles. For each gene, a newly created menu offers: 1) a tabular list of the variants that can be sorted by several parameters; 2) a gene graph providing a schematic representation of the variants along the gene; 3) statistical analysis of the data according to the phenotype, alteration type, and location of the mutation in the gene; 4) the cDNA and gDNA sequences of each gene, showing the nucleotide changes along the sequence, with a color-based code highlighting the gene domains, the first ATG, and the termination codon; and 5) a "download" menu making all tables and figures available for the users, which, except for the gene graphs, are all automatically generated and updated upon submission of the variants. Finally, the entire database was curated to comply with the HUGO Gene Nomenclature Committee (HGNC) and HGVS nomenclature guidelines, and wherever necessary, an informative note was provided. Infevers has already proven useful for the scientific community with a mean number of visits per month of 200 in 2002 and 800 in 2007, and its new design will lead to a more comprehensive comparative analysis and interpretation of auto-inflammatory sequence variants. PMID- 18409190 TI - Altered expression of 12S/MT-RNR1, MT-CO2/COX2, and MT-ATP6 mitochondrial genes in prostate cancer. AB - BACKGROUND: Prostate cancer is one of the commonest cancers worldwide and is responsible for nearly 6% of all male cancer deaths. Despite this relevance, the mechanisms involved in the development and progression of this malignancy remain unknown. The involvement of polypeptides of the mitochondrial respiratory chain, the Krebs cycle and the glutathione antioxidant system in this type of cancer has been previously described, although no publication has focused on the expression of mitochondrial genes in the prostate of PCa patients. METHODS: We have determined by reverse transcription-quantitative PCR (RT-qPCR) the relative amount of the transcripts of eight mitochondrial genes (MT-ND2, MT-ND4, MT-ND6, MT-CYB, 12S/MT-RNR1, 16S/MT-RNR2, MT-CO2/COX2, MT-ATP6), and four nuclear genes (COX11, GSR, CS, ACO2), all of them key players in the normal metabolism of mitochondria. Additionally we analyzed the expression of Cyclophilin A (PPIA). RESULTS: We observed differential expression of mitochondrial 12S/MT-RNR1, MT CO2/COX2, and MT-ATP6 transcripts in tumor samples when compared to their paired normal samples. CONCLUSIONS: The amount of mitochondrial 12S/MT-RNR1, MT CO2/COX2, and MT-ATP6 transcripts is significantly decreased in tumor samples when compared to their paired normal sample, suggesting that mitochondrial gene expression is altered in PCa. PMID- 18409193 TI - Weak self-association of human growth hormone investigated by nitrogen-15 NMR relaxation. AB - The self-association of human growth hormone(hGH) was investigated using 15N NMR relaxation.The investigation relies on the 15N R1 and R2 relaxation rates and the heteronuclear{1H}-15N NOEs of the backbone amide groups at multiple protein concentrations. It is shown that the rotational correlation time of hGH in solution depends strongly on its concentration, indicating a significant degree of self-association.The self-association is reversible and the monomers in the aggregates are noncovalently linked. Extrapolation of the relaxation data to zero concentration predicts a correlation time of 13.4 ns and a rotational diffusion anisotropy of 1.26 for monomeric hGH, in agreement with the rotational diffusion properties estimated by hydrodynamic calculations. Moreover, the extrapolation allows characterization of the backbone dynamics of monomeric hGH without interference from self-association phenomena, and it is found that hGH is considerably more flexible than originally thought. A concerted least-squares analysis of the 15N relaxations and their concentration dependence reveals that the self-association goes beyond a simple monomer-dimer equilibrium, and that tetramers or other multimeric states co-exist in fast exchange with the monomeric and dimeric hGH at sub-millimolar concentrations. Small changes in the 1H and 15N amide chemical shifts suggest that a region around the C-terminus is involved in the oligomer formation. PMID- 18409192 TI - Body mass index and magnetic resonance markers of brain integrity in adults. AB - OBJECTIVE: Obesity and being overweight during adulthood have been consistently linked to increased risk for development of dementia later in life, especially Alzheimer's disease. They have also been associated with cognitive dysfunction and brain structural alterations in otherwise healthy adults. Although proton magnetic resonance spectroscopy may distinguish between neuronal and glial components of the brain and may point to neurobiological mechanisms underlying brain atrophy and cognitive changes, no spectroscopic studies have yet assessed the relationships between adiposity and brain metabolites. METHODS: We have utilized magnetic resonance imaging and proton magnetic resonance spectroscopic imaging data from 50 healthy middle-aged participants (mean age, 41.7 +/- 8.5 years; 17 women), who were scanned as control subjects for another study. RESULTS: After adjustment for age and sex, greater body mass indices (BMIs) correlated with: (1) lower concentrations of N-acetylaspartate (spectroscopic marker of neuronal viability) in frontal (p = 0.001), parietal (p = 0.006), and temporal (p = 0.008) white matter; (2) lower N-acetylaspartate in frontal gray matter (p = 0.01); and (3) lower concentrations of choline-containing metabolites (associated with membrane metabolism) in frontal white matter (p = 0.05). INTERPRETATION: These results suggest that increased BMI at midlife is associated with neuronal and/or myelin abnormalities, primarily in the frontal lobe. Because white matter in the frontal lobes is more prone to the effects of aging than in other lobes, our results may reflect accelerated aging in individuals with high levels of adiposity. Thus, greater BMI may increase the odds of developing an age related disease, such as Alzheimer's disease. PMID- 18409194 TI - Three-dimensional model of the human urotensin-II receptor: docking of human urotensin-II and nonpeptide antagonists in the binding site and comparison with an antagonist pharmacophore model. AB - Human urotensin-II (hU-II) is a cyclic peptide that plays a central role in cardiovascular homeostasis and is considered to be the most potent mammalian vasoconstrictor identified to date. It is a natural ligand of the human urotensin II (hUT-II) receptor, a member of the family of rhodopsin-like G-protein-coupled receptors. To understand the molecular interactions of hU-II and certain antagonists with the hUT-II receptor, a model of the hUT-II receptor in an active conformation with all its connecting loops was constructed by homology modeling. The initial model was placed in a pre-equilibrated lipid bilayer and re equilibrated by several procedures of energy minimization and molecular dynamics simulations. Docking studies were performed for hU-II and for a series of nonpeptide hUT-II receptor antagonists in the active site of the modeled receptor structure. Results of the hU-II docking study are in agreement with our previous work and with experimental data showing the contribution of the extracellular loops II and III to ligand recognition. The docking of hU-II nonpeptide antagonists allows identification of key molecular interactions and confirms a previously reported hU-II antagonist pharmacophore model. The results of the present studies will be used in structure-based drug design for developing novel antagonists for the hUT-II receptor. PMID- 18409195 TI - Chronic toxic encephalopathy caused by occupational solvent exposure. PMID- 18409196 TI - Inclusions of amyotrophic lateral sclerosis-linked superoxide dismutase in ventral horns, liver, and kidney. AB - Mutant superoxide dismutases type 1 (SOD1s) cause amyotrophic lateral sclerosis by an unidentified toxic property. In a patient carrying the G127X truncation mutation, minute amounts of SOD1 were found in ventral horns using a mutant specific antibody. Still, both absolute levels and ratios versus wild-type SOD1 were considerably greater than in other central nervous system areas and peripheral organs. Inclusions of mutant SOD1 were abundant in motoneurons but were also seen in hepatocytes and kidney epithelium. This first examination of mutant SOD1 in both central nervous system and peripheral organs supports the notion that enrichment of misfolded SOD1s might explain the particular vulnerability of motor areas. PMID- 18409198 TI - Genetic mimics of congenital lymphocytic choriomeningitis virus encephalitis. PMID- 18409199 TI - Superoxide dismutase may lead to cytoskeletal abnormalities and amyotrophic lateral sclerosis. PMID- 18409200 TI - High risk studies and developmental antecedents of anxiety disorders. AB - The past two decades have witnessed significant growth in our understanding of the developmental antecedents of anxiety disorders. In this article, we review studies of offspring at risk for anxiety disorders, longitudinal studies of the course of anxiety disorders in clinical, epidemiologic, and at-risk samples, studies of hypothesized temperamental risk factors for anxiety, and give a brief overview of the literature on environmental risk factors. Clear developmental antecedents to anxiety disorders identified include (1) childhood anxiety disorders [in particular, separation anxiety and overanxious disorder/general anxiety disorder (GAD)], (2) behavioral inhibition which predicts later social phobia, (3) anxiety sensitivity which predicts later panic disorder, and (4) negative affectivity, which predicts a spectrum of psychopathology including anxiety disorders. Further prospective studies are needed to examine the roles of environmental factors such as parenting practices, peer influences, stressful life events, and perinatal stressors. Future studies could benefit from (1) beginning earlier in development and following individuals into adulthood, (2) assessing the overlap between multiple temperamental constructs, (3) greater use of observational measures of temperament and of parent-child and peer interactions, (4) greater attention to parental psychopathology which may confound associations noted, (5) exploration of other features of anxiety disorders (neurofunctional correlates, cognitive features, other aspects of emotional regulation) as potential precursors, and (6) intervention studies exploring whether modifying developmental antecedents can alter the course of anxiety disorders. PMID- 18409201 TI - No major role for the EMX2 gene in schizencephaly. AB - Schizencephaly (SCH) is a rare disorder of cerebral cortical development, characterized by full thickness clefts spanning the wall of the cerebral hemispheres that are lined and surrounded by polymicrogyric cortex. Based on pathological analysis, SCH was originally considered to have multiple causes including infectious and vascular injuries, and toxic agents. However, a few reports of familial SCH have suggested a possible genetic etiology. Ten years ago two articles identified EMX2 as the first causative gene for human SCH in 13 of 18 patients, although for several putative mutations no pathogenic role was demonstrated. Here, we reinterpret the original articles as showing a significantly lower mutational rate (17%) than originally reported (72%), and provide results of EMX2 sequencing in 39 new SCH patients, detecting no pathogenic mutations. We conclude that the reported association between SCH and EMX2 mutations is not adequately supported by current data, and that diagnostic testing of EMX2 is not justified, as any results would be uninterpretable. PMID- 18409202 TI - Compound heterozygosity for two MSH6 mutations in a patient with early onset colorectal cancer, vitiligo and systemic lupus erythematosus. AB - Lynch syndrome (hereditary non-polyposis colorectal cancer, HNPCC) is an autosomal dominant condition caused by heterozygous germline mutations in the DNA mismatch repair (MMR) genes MLH1, MSH2, MSH6, or PMS2. Rare cases have been reported of an inherited bi-allelic deficiency of MMR genes, associated with multiple cafe-au-lait spots, early onset CNS tumors, hematological malignancies, and early onset gastrointestinal neoplasia. We report on a patient with vitiligo in segments of the integument who developed systemic lupus erythematosus (SLE) at the age of 16, and four synchronous colorectal cancers at age 17 years. Examination of the colorectal cancer tissue showed high microsatellite instability (MSI-H) and an exclusive loss of expression of the MSH6 protein. Immunohistochemical analysis of normal colon tissue also showed loss of MSH6, pointing to a bi-allelic MSH6 mutation. Sequencing of the MSH6 gene showed the two germline mutations; c.1806_1809delAAAG;p.Glu604LeufsX5 and c.3226C > T;p.Arg1076Cys. We confirmed that the two mutations are on two different alleles by allele-specific PCR. To our knowledge, neither parent is clinically affected. They did not wish to be tested for the mutations identified in their daughter. These data suggest that bi-allelic mutations of one of the MMR genes should be considered in patients who develop early-onset multiple HNPCC-associated tumors and autoimmune disorders, even in absence of either hematological malignancies or brain tumors. PMID- 18409203 TI - The arthrochalasia type of Ehlers-Danlos syndrome (EDS VIIA and VIIB): the diagnostic value of collagen fibril ultrastructure. PMID- 18409204 TI - Revisiting Wittgenstein on Kohler and Gestalt psychology. AB - In an article in this journal, Nicholas Pastore rejected Ludwig Wittgenstein's critique of Wolfgang Kohler and Gestalt psychology. Pastore appears not to have appreciated Wittgenstein's argument that Kohler mistook conceptual questions for factual ones. A simi-lar confusion seems to underlie at least some aspects of contemporary neuroscience. Be that as it may, Wittgenstein has had minimal influence on the research practices of psychologists while Kohler remains influential. This outcome would not have surprised Wittgenstein, who predicted that scientists would not see his work as relevant to theirs. PMID- 18409205 TI - From Vygotsky to Vygotskian psychology: introduction to the history of the Kharkov School. AB - Around the end of the 1920s, Vygotsky introduced his integrative framework for psycho-logical research to the Soviet Union. This framework was not abandoned and forgotten until its rediscovery in Russia and America in the 1950s, as some claim. In fact, even after his untimely death in 1934, Vygotsky remained the spiritual leader of a group of his for-mer students and collaborators, who became known as the Kharkov School. This paper reconstructs the early intellectual history of Vygotskian psychology, as it emerged, around the time of Vygotsky's death, in the research program of the Kharkov School. PMID- 18409206 TI - Spanish experience with German psychology prior to World War I. AB - An increase in interest for German scientific psychology followed the rise of liberalism in late nineteenth-century Spain. This paper deals with Spanish scholars' endeavors to participate in German psychology: It outlines the intellectual and institutional background of Spanish preoccupation with German philosophy and psychology, and deals with the personal experience and testimony of two Spanish philosophers, Eloy Luis Andre and Juan Vicente Viqueira Lopez, who traveled to Leipzig, Berlin, and Gottingen between 1909 and 1914 to gain firsthand experience in the nascent science of psychology in Germany at that time. PMID- 18409207 TI - Toward a transnational history of the social sciences. AB - Historical accounts of the social sciences have too often accepted local or national institutions as a self-evident framework of analysis, instead of considering them as being embedded in transnational relations of various kinds. Evolving patterns of transnational mobility and exchange cut through the neat distinction between the local, the national, and the inter-national, and thus represent an essential component in the dynamics of the social sciences, as well as a fruitful perspective for rethinking their historical development. In this programmatic outline, it is argued that a transnational history of the social sciences may be fruitfully understood on the basis of three general mechanisms, which have structured the transnational flows of people and ideas in decisive ways: (a) the functioning of international scholarly institutions, (b) the transnational mobility of scholars, and (c) the politics of trans-national exchange of nonacademic institutions. The article subsequently examines and illustrates each of these mechanisms. PMID- 18409208 TI - Closing the gaps. PMID- 18409209 TI - A simple model for metal cation-phosphate interactions in nucleic acids in the gas phase: alkali metal cations and trimethyl phosphate. AB - Threshold collision-induced dissociation techniques are employed to determine the bond dissociation energies (BDEs) of complexes of alkali metal cations to trimethyl phosphate, TMP. Endothermic loss of the intact TMP ligand is the only dissociation pathway observed for all complexes. Theoretical calculations at the B3LYP/6-31G* level of theory are used to determine the structures, vibrational frequencies, and rotational constants of neutral TMP and the M+(TMP) complexes. Theoretical BDEs are determined from single point energy calculations at the B3LYP/6-311+G(2d,2p) level using the B3LYP/6-31G* optimized geometries. The agreement between theory and experiment is reasonably good for all complexes except Li+(TMP). The absolute M+-(TMP) BDEs are found to decrease monotonically as the size of the alkali metal cation increases. No activated dissociation was observed for alkali metal cation binding to TMP. The binding of alkali metal cations to TMP is compared with that to acetone and methanol. PMID- 18409210 TI - The role of local biosynthesis of auxin and cytokinin in plant development. AB - Plant hormones are tightly regulated in response to environmental and developmental signals. It has long been speculated that biosynthesis of hormones occurs broadly in plant organs and that intricate, spatiotemporal regulation of hormones in developing organ primordia is achieved through transport and signal perception. However, recent identification of genes crucial for biosynthesis of auxin and cytokinin reveals that localized hormone biosynthesis also plays an important role in organ growth and patterning. PMID- 18409211 TI - A quaternary question. Response. PMID- 18409212 TI - [Methods of fetal surveillance during labor. Guidelines]. PMID- 18409214 TI - Introspections. Joy of caring. PMID- 18409213 TI - [Prevention of cervical cancer. Guidelines]. PMID- 18409215 TI - [GKV-WSG--what is the status today?]. PMID- 18409216 TI - [What type of surgeon does the surgical practice of the future need?]. PMID- 18409217 TI - [Revised regulations in German federal contract physicians and national insurance comprehensive professional practice guideline subsequent to the modified contract physician legal directive]. PMID- 18409218 TI - [Basic aspects of authorization of hospital physicians for participating in contract physician health care]. PMID- 18409219 TI - [Surgeons in Norway]. PMID- 18409220 TI - [Reorganization of ambulatory care--benefit for the family physician, specialist and patient ]. PMID- 18409221 TI - [Professional taxation problems in physician enterprises]. PMID- 18409222 TI - [Sterilization of medical products in the physician's practice]. PMID- 18409223 TI - [DRG practice: osteosynthesis of fractures--what are material combinations?]. PMID- 18409224 TI - [Communication with patients]. PMID- 18409225 TI - [Classification of physician directors]. PMID- 18409226 TI - [Physician treatment error and legal ramifications] ]. PMID- 18409227 TI - A new technique of inguinal hernia repair--neither similar to nor modification of Desarda's repair. PMID- 18409228 TI - Formation of Indian Medical Federation. PMID- 18409229 TI - Snakebite management in India. PMID- 18409230 TI - Snakebite management in India. PMID- 18409231 TI - Generic drugs. PMID- 18409232 TI - Re. Operation Blessing in New Orleans: PDA president Johnston and president-elect Gamba reach out to help. PMID- 18409233 TI - A comparative review of periodontal and peri-implant soft tissue. Part 2. PMID- 18409234 TI - Cardiovascular diabetology: clinical, metabolic and inflammatory facets. Preface. PMID- 18409235 TI - Re: in memoriam: Ramon James Evans (1932-2007). Pain Res Manag 2007; 12:244. PMID- 18409236 TI - Possible roles of endogenous plant viral sequences and transgenes containing viral sequences in both virus resistance and virus emergence. PMID- 18409237 TI - Aim and contents--1st International Symposium on Green Technology and Ecosystem Health, Hangzhou, China, Oct. 13-15, 2007. Preface. PMID- 18409238 TI - Obesity and metabolism. Preface. PMID- 18409239 TI - E-prescribing. PMID- 18409241 TI - Rethinking informed consent. PMID- 18409240 TI - Smoke-free laws. PMID- 18409243 TI - Louisiana State Board of Dentistry update. PMID- 18409242 TI - [The cardiology nurse at the heart of care]. PMID- 18409244 TI - "Tap into a healthier smile" fluoridation initiative will need grassroots advocacy. PMID- 18409245 TI - The 5 deadly misconceptions with 6 links of survival: are you truly prepared for medical emergencies? PMID- 18409246 TI - The removable partial denture: is it irrelevant in dental school curriculum? PMID- 18409247 TI - Finding direction for senior oral health. PMID- 18409248 TI - Florida's dentists making a difference. Broadening access to oral health care. PMID- 18409249 TI - Buker marks 20 years at the FDA. PMID- 18409250 TI - The true last years and legacy of Dr. John Hinchman Stokes. PMID- 18409251 TI - Community health workers. PMID- 18409252 TI - Osteoporosis breaks bones and budgets. PMID- 18409253 TI - Putting wound-related pain on the health-care agenda. PMID- 18409254 TI - Assessment of wound pain: overview and a new initiative. AB - All wounds have the potential to cause pain, and the nature of the pain varies with the type of wound. Many factors may exacerbate wound pain, including infection, trauma at dressing changes and poor technique when applying compression therapy. Failure to assess wound pain or inadequate pain assessment can cause the patient further anguish and extended suffering. Nurses caring for patients with painful wounds need to identify the source of the pain and exacerbating factors, and determine whether it has nocicoceptive and/or neuropathic elements in order to optimize pain management for the individual patient. This article examines the assessment of wound pain and introduces an initiative that has been developed to improve the assessment process. The 'Heal not Hurt' initiative is an excellent example of the profession and industry working together to implement best practice guidance in patient-centered pain free wound care in clinical care. PMID- 18409255 TI - 'Heal not Hurt': piloting an intitiative on wound pain assessment. AB - As a number of consensus and position documents have made clear, wound pain is an issue that all nurses involved in wound care should take seriously. Based on the 'Heal not Hurt initiative supported by Molnlycke, a wound assessment tool has been adopted by East Riding of Yorkshire PCT. This article gives an overview of the documentation and argues that thorough assessment is essential in wound care and proper documentation is the key to successful clinical outcomes. PMID- 18409256 TI - The cost of reducing pain associated with chronic wound management. AB - The significant financial costs associated with care of chronic wounds are commonly cited, but health economic analyses are often undermined by incomplete data and poor evidence of the effectiveness of interventions. This article highlights additional elements of cost associated with wounds that are rarely if ever measured: the human costs. In discussing recent studies into this area, it concludes that patient priorities may differ from those of health professionals and budget holders, but must be considered an essential component of future cost studies. PMID- 18409257 TI - [IARC evaluations: how are they conducted and what suggestions are derived therefrom?]. AB - The IARC Monographs on the Evaluation of Carcinogenic Risks to Humans are considered by several Institutions a point of reference for the prevention of adverse effects (namely cancer) of occupational exposures. The conclusions which lead to the Classification of a substance or an industrial process are the results of a vote within a Working Group of experts and are expressed in terms of "evidence" based on both epidemiological and experimental data or "other relevant data". Several points are discussed which include the methods of identification of the experts, the criteria of evaluation of the available data, the meaning of some definitions and finally the impact of IARC conclusions and Classification on Public Health regulations and secondarily on compensation procedures or trial decisions. Three recent examples regarding the Classification of Free Crystalline Silica, Shift-Work and Vinyl Chloride Monomer, which cannot be accepted as such without some criticism, are reported. In conclusion the Authors appreciate the effort of the IARC to provide a source of information which "may assist national and international authorities in making risk assessments and in formulating decisions concerning any necessary preventive measures". However, they suggest that the IARC "overall evaluation" should not be taken as confidently as if no doubt at all would exist on the final statement and the consequent Classification. Some suggestions as to the solution of the question are also provided. PMID- 18409258 TI - [Chemical risk in health environment: contribution to the evaluation and management of the risk to health]. AB - Among the many workplaces where there is a chemical risk, hospitals are surely one of the most complex when it comes to risk assessment. Numerous highly toxic substances are routinely used; clinical decisions often mean health workers come into contact with latest-generation drugs whose secondary effects are not yet fully known; they may have to execute procedures using different chemicals at the same time, as often happens in therapy or surgery. We studied two provincial hospitals in order to make a preliminary risk assessment, using an algorithm model. The chemical risk in the environments analyzed was mainly moderate, though some specific settings call for closer attention. Certain at-risk conditions can be solved by simple procedural changes, but others need to be tackled with a view to residual risk management. Some critical points came to light, linked to the algorithm employed, which need further examination to fit the model better to this particular context. PMID- 18409259 TI - [Collateral effects of a project of latex rubber removal in a hospital institution]. AB - Within a stepwise approach to remove natural rubber latex products from the hospital of Cuneo, latex gloves for examination were completely replaced by nitrile powder free gloves. About a month after their introduction in the whole facility, 31 health care workers (HCW) presented with symptoms of dermatitis characterized by erythema, itching, dryness and chapping on the dorsum of the hands. The symptoms typically appeared 1-3 days after the first nitrile gloves employment, 1-3 hours after they were put on. On the basis of both a detailed clinical examination and specific laboratory tests a diagnosis of irritant contact dermatitis was confirmed in all symptomatic HCW. Samples of nitrile gloves were submitted for analysis by a university toxicological laboratory. Evident traces of a powder containing calcium carbonate were found inside of the gloves. Therefore the declaration of the firm that the product was absolutely powder free must be considered wrong. We suppose that frequent and prolonged contacts between the alcaline pH of the powder found in the gloves and the acid pH of the skin of the hands - together with other individual and environmental factors - could have determined the onset of the symptoms in the HCW. In spite of this adverse reaction, a significant reduction (-68%) of the incidence of latex sensitization and latex-related symptoms was observed in 2006 compared to 2005. We conclude that the complete removal of latex gloves from the workplace is a favourable preventive measure and that switching to nitrile powder free devices is a safe and cost-effective alternative to the latex gloves. However, it is always advisable to demand to the producer a formal declaration of compliance to the security standards of the medical devices. PMID- 18409260 TI - [Occupational exposure to environmental pollution: observational study about a newspaper-agents group]. AB - Environmental pollution constantly menaces urban people's health. This public health question has a remarkable interest in occupational medicine, because some professions require operations outdoor for most of the day. Therefore airborne pollutants could be considered occupational risk factors. Our object is health evaluation of a newspaper-agents group of Palermo, in association with pollution level monitored during the 2006. We examined these subjects regarding especially lung, ear, kidney, liver and cardiovascular system, with particular consideration for endothelial structure. RESULTS: high prevalence of carotid artery damage can't exclude a concausal role of airborne pollutants. PMID- 18409261 TI - [Report on the incidence of accidents and occupational diseases in the sector of waste disposal]. AB - Waste management is continuously evolving, moreover, lack of resources and environmental hygiene issues have given more importance to recycling, establishing the basis of an increased risk in these workers. The aim of this study was to provide a description of occupational health of Italian waste workers in order to highlight key points for ad hoc interventions. Data about injuries and professional disease were extracted from the Italian National Labour Insurance Institute (INAIL) Waste work is a well-known cause of occupational accidents, however occupational disease appear to be less-known with many "unclassified" events. While new researches are needed in order to clarify these emerging issues, education could be the key to assure the waste workers health safety. PMID- 18409262 TI - Lung cancer in subjects suffering from silicosis in the Province of Genoa from 1979 to 2004. AB - The Authors, after a survey of the latest experimental and epidemiological studies concerning the possible role of crystalline silica in lung carcinogenicity, examine the cases acknowledged by the INAIL in Genoa concerning subjects suffering from silicosis who, thereafter, developed a lung cancer in the period 1979 - 2004. Distribution of the degree of disability, of the length of occupational exposure to silica, of the workers' age at the time of death and of the different fields of activity have been tested; smoking habits and histological classification of the neoplasms have also been assessed, when available. Our data seem to agree with the point of view of the Authors who so far have expressed perplexities about the role played by silica in lung carcinogenesis, especially if associated with the exposure to powerful carcinogenic agents such as cigarette smoke. At present we share the opinion of those who think that a final regulation should be delayed up to the time when it will be possible to identify the occupational source of silica with such characteristic as to make it actually carcinogenic. PMID- 18409263 TI - [Mental health and life style changes in young workers]. AB - Lifestyle's alterations are hazardous for health. On one hand they produce a high rate of mortality and disease, on the other hand they cause a reduction of work outcomes and an increase of occupational accidents with important consequences for both worker's health and his/her financial status. The aim of this study is to review the scientific literature for possible relationships between mental health and lifestyle alterations of young workers. We considered as lifestyle factors the attitudes towards: smoking, alcohol consumption, eating, use of medications and doping substances, physical activity and sleeping. From our study it clearly emerges the existence of correlation between lifestyle habits and mental health; in fact behavioural alterations can produce problems of mental health and vice versa. Furthermore, some work peculiarities can lead to psychic disturbances and/or to unhealthy habits which can themselves cause negative effects on working activity. It is very important for young workers to understand that unhealthy behaviours, which can be corrected, are hazardous in terms of health and safety for both the single worker and the collectivity and that those behaviours can enhances the other working risks. Because there is a close interaction between mental health and lifestyles, it would be necessary a careful promotion of mental health on workplaces and to take all the preventive measures, with particular regard for those related to the work organization, in order to reduce the onset, exacerbation and unmasking of mental disorders and psychological difficulties. In working environment, the occupational health physician and his relationship with the patient are of fundamental importance. During preventive and periodic medical examinations, the occupational health physician should take detailed information on young worker's habits regarding smoking, alcohol consumption, eating, physical activity, sleeping, pharmacological abuse and possible presence of mental disorders and furthermore he/she should actively take part in the information and education process of the worker. PMID- 18409264 TI - [Forensic implications and stress ina group of anaestheticians: a cross sectional study]. AB - Aim of the study is the appraisal of stress in a group of the 500 anaestheticians and in a control group. Two anonymous self-administered questionnaires were utilized, one for the subjective appraisal of the perceived various risk related to job, and one for the subjective appraisal of stress: VRS questionnaire of Tarsitani and Biondi. The results of the study showed that all the anaestheticians identified the perception of the emotional aspects related to work as a risk factor. They put it mainly in relation with the forensic implications. The total score of scale VRS and the score for cluster depression, anxiety, aggressiveness are higher in the anesthetists than the control group. PMID- 18409265 TI - [Occupational backache: crossover study]. AB - The authors briefly review the risk factors for the spine involved in working activities. Three hundred workers doing different jobs were studied for their general physical condition, job-related postures, the site of any back pain, and motor activity practiced. In line with other studies, data confirm that no worker category is immune from back problems, but highlight the positive role of motor activity in pain perception. PMID- 18409266 TI - [Risk factors for musculoskeletal complaints among microscope workers]. AB - OBJECTIVES: To describe the prevalence of musculoskeletal complaints among microscope workers, and to investigate the relationship between complaints in different body regions and individual as well work-related factors. METHODS: A detailed questionnaire on musculoskeletal symptoms was distributed to 176 microscope workers. The questionnaire contained questions on individual factors, workstation characteristics, work organization and psychosocial factors. RESULTS: The 1-month prevalence of symptoms at any body site was 76,1%. Symptoms were most commonly reported at the neck (53,4%), followed by the lower back (39,2%), upper back (23,9%), shoulders (21%), hand/wrist (21%) and arms (6,3%). Female gender was associated with complaints in all sites except low back. Hours of microscope use was associated with neck, arm and hand/wrist pain, whereas duration of work time without breaks was associated with neck, low back, and arm pain. Poor workstation ergonomics was associated with symptoms in the neck, shoulders, upper and lower back. Work pace was also identified as a risk factor for low back complaints. CONCLUSION: This study supports the need for ergonomic intervention at workplace and ergonomic training of microscope workers. PMID- 18409267 TI - [Industrial manufacturing of the wool: pathology secondary to the biomechanical involvement of the upper limbs and the course in relation to work tasks]. AB - In years 2003-2005, 1248 laborers of Zegna Baruffa - Lane Borgosesia Company (establishment of Borgosesia and Vallemosso) have been visited and we selected 102 cases of pathology from upper extremity work-related musculoskeletal disorders in the units with working risk. Subsequently from April 2005 to May 2006 we have controlled the clinical evolution of the enlisted laborers and we noted that in 54% of the cases the clinical symptomatology has been resolved with a rest works and a correct therapy. The diagnosis, the treatment and the change of working duty can resolve a meaningful number of clinical cases. PMID- 18409268 TI - [Cycle ergometers in rehabilitation medicine: technical characteristics and selection criteria]. AB - The cycle ergometer is a commonly used means of testing and training patients with energetic disabilities (particularly in the area of cardiopulmonary diseases). Recent evidence suggests that cycle ergometers can also be useful in patients with subacute and chronic stroke, brain injury, chronic degenerative neurological conditions, and in spinal cord injury. Commercially available cycle ergometers show wide differences in terms of structure and function that have a direct impact on the specific rehabilitation protocols. The aims of this review paper are: a) to briefly review the physiology and biomechanics of exercise on a cycle ergometer; b) to review and discuss the technical specifications of the cycle ergometers suited to rehabilitation settings; c) to provide guidelines for selecting appropriate cycle ergometers for the different categories of rehabilitation patients. First, the physiology of exercise on the cycle ergometer and its biomechanical features are discussed, including the patterns of muscular activity during down- and up-stroke. Upright and recumbent ergometers and their specific clinical indications are compared. Finally, the technical characteristics of the cycle ergometers (load, motor, resistance, flywheel, belt, resilience, pedals, frame, display) are described and the specific requisites for the different patient categories undergoing rehabilitation are discussed in detail. Finally, guidelines are offered for identifying the main technical requisites for appropriate cycle ergometer selection in the different disabilities. PMID- 18409269 TI - Smoking in Hollywood movies: impact on teen smoking with special reference to German adolescents. AB - This paper summarizes studies that have linked exposure to movie smoking and smoking initiation among adolescents. Much of the research linking exposure to smoking to movies with adolescent smoking comes from studies of U.S. children and their exposure to smoking in Hollywood movies. Cross-sectional and longitudinal studies have assessed such exposure and have found a strong, independent association with smoking onset. A first study conduced in Germany reveals that smoking in internationally distributed movies is a risk factor for ever and current smoking among European adolescents, too. It is concluded that limiting exposure of young adolescents to movie smoking could have important world-wide public health implications. PMID- 18409270 TI - [Maternal smoking: effect on neonatal's health]. AB - The influence of maternal smoking on the higher miscarriage rate, premature births and premature detachment of the placenta was proved in numerous so far undertaken medical researches. Apart from that smoking has an influence on fetal developmental retardation, low birth weight as well as the baby's abnormal development. The investigated population comprised of 100 healthy women that gave birth to their children after the 37th week of pregnancy and their newborns. The pregnant women's morphology parameters, the newborns' health (after the third minute of life according to the Apgar's scale) and the birth weight were assessed. The results achieved were analysed taking into consideration the problem of maternal smoking. The lower Apgar's notes as well as lower birth weight of the newborns whose mothers used to smoke during pregnancy were observed. The fact of being pregnant is the most significant argument for women to give up or diminish the habit of smoking. PMID- 18409271 TI - [Socio-economic determinants of smoking during pregnancy and birth weight of newborns]. AB - In this survey, we have assessed a socio-economic status of pregnant women smoking during pregnancy and the impact of this status on birth weight of their newborns. The research, we have done on the group of 431 pregnant women. Smoking habit has been declared by 21.6% of women and passive exposure to tobacco smoke has been stated by 32.5% of respondents. We have observed a significant association between active and passive smoking and socio-economic status of respondents. The newborns of actively smoking mothers were lighter of about 216 g according to those ones of mothers who did not smoke and were not exposed to passive smoking, and were lighter than newborns of mothers who were not exposed to passive smoking of about 146 g. We have not observed any significant link between active and passive exposure to tobacco smoke during pregnancy, and a risk of delivering low weighted child. However, in case of newborns weighted above 2500 g., their mothers have not been smoking and have not been exposed to tobacco smoke significantly more often. PMID- 18409272 TI - [Passive smoking at teenager's home suffering from asthma bronchiale and control at the disease with Asthma Control Test]. AB - Allergies of the respiratory system are very often at children. Passive smoking may predispose to allergies. The last news shows that smoking at home increases incidence of asthma. It's concerned passive smokers, especially children. Analysis of controlling asthma, exacerbations was made with ACT test. The larger amount of points in ACT test was obtain the better control of asthma was ascertain. Parents were smokers. In this research 164 children and teenagers between 12 and 18 years old from Swiecie and Bydgoszcz, patients Allergologic Outpatients Clinic in 2005-2006, took part. Asthma Control Test was made after the treatment was started. This results show that there is a strong dependence between smoking at home and controlling asthma bronchiale. The correct score--25 points (means that asthma was controlled properly) was at 75% of patients with no smoking at home. At smoker's home every second child has controlled the disease properly. In families where parents smoke a lot every fifth child has no control of the disease. In families, where parents didn't smoke it was only 3%. This analysis shows that there is a strong dependence between frequency of smoking and amounts of points in ACT test. In families where parents smoke rarely children received maximum score in ACT test than in families where patients smoke a lot. There is also dependence between the age of the members of the household and smoking. It is terrifying that there are far more smoking parents at the age of 35 and younger. In this study the ACT was recognized as useful test to control asthma. If smoking parents undergo antinicotine therapy treatment of asthma will be improved. PMID- 18409273 TI - [Asthma bronchial among children and youth exposed to tobacco smoke]. AB - Purpose of this work was to judge progress of diagnosed bronchial asthma among children and youth that are exposed on tobacco smoke in aspect of number of exacerbation of disease, therapy (doses of corticosteroids) and number of hospitalizations. There were two groups of patients with bronchial asthma under examination, first one were children and youth that are exposed on tobacco smoke, and the other group of children and youth that are not exposed on tobacco smoke. We discovered statistically significant difference in number of exacerbation of disease among those two groups, also bigger therapeutic and prophylactic doses of inhaled steroids during treatment, and much more often hospitalizations among children and youth that are exposed on tobacco smoke. PMID- 18409274 TI - Consequences of passive smoking in home environment. AB - Passive smoking means cigarette smoke inhaling by people other than smokers. Passive smoker inhales tobacco smoke coming not only from side-stream, but also smoke exhaled by the smoker. Long-term tobacco smoke inhaling increases the risk of appearance of smoke related diseases (for example COPD, heart diseases), including the most dangerous types of cancer, which only few smokers realize. The aim of this study was to check whether tobacco smoke inhaling in home environment from childhood to adulthood has an influence on respiratory system of adults. The study included adults. In the study two types of participants division were used. Among 1481 persons two groups were separated. Group 1 contains people, who have never lived in home environment with active smokers, altogether 465 persons. Group 2 contains people who from birth have been exposed to cigarette smoke inhaling (altogether 1016 persons). With help of the author's questionnaire the information concerning demographic features and smoking habit were gathered. The patients underwent doctor's examination. They had a spirometric test and a chest radiogram. Statistically significant differences appear among persons with higher education who belong to two different groups. The number of children who have lived in smoke free rooms during childhood and adolescence and finished studies is bigger than the number of active smokers' children (p<0.001). Over 90% of never smokers have inhaled cigarette smoke since childhood in home environment. Chronic obstructive pulmonary disease has been diagnosed at 47.5% of active smokers, 48.3% of ex-smokers and up to 44.7% of passive smokers. Cigarette smoking and smoke inhaling for more than twenty years has a significant influence on the appearance of COPD. No statistically significant differences were noted among the three groups of participants. (p>0.05). Cigarette smoke inhaling at childhood and adolescence should be taken seriously because it causes development of chronic diseases like COPD. GOLD 2006 standards convince that at every stage of COPD development the effects of this disease may be partly reversed if one stops smoking cigarettes. That is why a child should not be exposed to cigarette smoke at any stage of its life. Permanent inhalation of tobacco smoke since early childhood in home environment influences equal occurrence of COPD at passive, ex- and active smokers. Reduction of tobacco consumption, better care concerning passive smokers, increasing consciousness of parents smoking in home environment may protect their children from serious health consequences in the future. PMID- 18409275 TI - [Analysis of the influence of nicotynism on selected diseases of the head and neck]. AB - Cigarette smoking is still an actual problem. Poland has a leading position in consumption of cigarettes. The aim of this research is analysis of morbidity risk factors of selected neck and head diseases. In our study we examined 387 patients. Among these patients were 187 patients with malignant tumors, 68 with pre-cancer diseases, 40 with nonmalignant neoplasms of the salivary gland and 92 patients with chronic sinusitis. PMID- 18409276 TI - [The impact of smoking cigarettes on insomnia and excessive daytime sleepiness symptoms among teachers]. AB - One of the most important problems in sleep medicine is Sleep Related Breathing Disorder and Insomnia. The excessive daytime sleepiness is basic symptom of those disorders. In this study we analyze some parameters of insomnia severity and excessive daytime sleepiness among smoking and nonsmoking teachers. In this study we used the own questionnaire about smoking, and Athens Insomnia Scale; Insomnia Severity Index; Epworth Sleepiness Scale; Fatigue Severity Scale. The 94 teachers was included to the study, 99% was female. We don't observe any differences between smoking and no-smoking group in any parameters. In the next step we categorize results and there no significant differences between frequency in both groups. There no significant differences between in both groups even we include persons who smoking in the past. Additional studies are needed to describe the connection between smoking cigarettes and sleep problem in different aged group. PMID- 18409277 TI - [Tobacco smoking among hospitalised patients with arterial hypertension]. AB - Currently 33% of the Polish adult population smoke tobacco (42% males and 25% females), which is a serious issue, particularly in the context of cardiovascular diseases. A study of the extent of smoking among patients with arterial hypertension (AH) served as the basis for the present article. The studied group comprised 100 patients hospitalised due to AH, 46 males and 54 females. A survey, aided by a specially designed questionnaire, was the method of the medical part of the study. It concentrated on health-related behaviour patterns conducive, or not, to AH treatment such as: systematic control of blood pressure, proper diet, physical activity, and tobacco smoking. The respondents were also asked to indicate the sources of their knowledge on the proper, in this context, lifestyle. The study revealed that 46.0% of the subjects smoked; most of them males, inhabitants of a large city (both genders), over 50-year-old, and with secondary school education. The majority of the group, while declaratively fully aware, thanks to the received information, of the necessity of the cessation of smoking - fails to react appropriately. Tobacco smoking constitutes a vital issue among the AH patients. The physician is not always the source of knowledge on the threats, associated with the addiction. The fact that 2/3 of the patients aware of the necessity of smoking cessation fails to do so is a reason for concern. PMID- 18409278 TI - [Chosen determinants of smoking among persons with rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) is a chronic connective tissue disease with characteristic symmetric joints infibulations leading to disability. Due to progressive character of the disease, with periods of worsening and remission, the treatment should be complex and focused on the inhibition of the progress of the changes. The aim of the study was opinion of effect of smoking for functionality in person with rheumatoid arthritis. Material consists of 23 persons with RA. HAQ Disability Score were used. Additionally the questionnaire for estimation of health attitudes was used. Results of investigations prove lack of connections between smoking and correct functionality in persons with rheumatoid arthritis. PMID- 18409279 TI - [Influence of tobacco smoking on physical efficiency of young woman (part I)]. AB - For over 20 years Poland has been one of the countries with the world's highest consumption of tobacco. In view of the widespread popularity of this addition, of particular importance becomes the problem of women becoming addicted to tobacco smoke. Therefore, the objective of this work was to develop and conduct preliminary surveys aimed at the determination of the scale of active and passive cigarette smoking among young women as well as the assessment of smoking on physical fitness. The research took a form of an anonymous questionnaire administered to 80 female secondary school graduates who were passing their entrance examinations to the University of Physical Education in Wroclaw. The physical capacities were assessed on the basis of fitness tests and their results were subject to statistical analysis. One identified a significant impact of cigarette smoking on reducing the physical fitness and, in consequence, lower scores in the fitness test among smoking girls when compared to the scores of non smoking girls (respectively 17.5+/-2.0; 21.0+/-4.5; p<0.0001). It was shown that smoking more than 10 cigarettes per day and on the fitness test day has considerably influenced their physical fitness (p<0.002; p<0.0004). Moreover, smoking women devoted less time to exercises and had a higher BMI when compared to non-smoking women. PMID- 18409280 TI - [Influence of tobacco smoking on physical efficiency of young men (part II)]. AB - Cigarette smoking by the young is becoming an ever more serious problem. It has been found that starting smoking at a younger age entails higher risks than when doing so at an older age. The life expectancy of people who take up this addiction before the age of 15 was demonstrated to be 8 years shorter. Research indicates that smoking considerably reduces physical fitness of young people. Therefore, the objective of this work was to assess the influence of smoking on the physical fitness of secondary school graduates passing their entrance examination to the University of Physical Education on the basis of surveys administered to them. The results were subject to statistical analysis. One identified a considerable influence of the number of cigarettes smoked per day and smoked on the examination day on the score earned during fitness tests (p<0.0003; p<0.005). It was also demonstrated that smoking youth devoted less time to the weekly physical workout than non-smoking--which also had its adverse impact on the test results. Non-smoking men did their workout 7.3+/-2.5 hours per day on average, while smoking men--4.6+/-3.2 hours (p<0.0001). These data confirm the fact that along with an increase in the number of cigarettes smoked and the addiction duration, the physical fitness of smokers goes down. PMID- 18409281 TI - [Analysis of dietary habits and dietary intake in hypertensive smokers]. AB - The aim of this study was to compare dietary habits and nutrient intake of hypertensive smokers with hypertensive non-smokers. The study population comprised 30 hypertensive smokers and 35 non-smokers, aged 30-60y. The participants were subjected to a triple 24-h diet recall and to a dietary habits questionnaire. The results showed that smokers had unhealthy patterns of nutrient intake. Smokers declared consuming too much fat, cholesterol and too low antioxidant vitamins, calcium and fibre. It was found that concentration of vitamin E in smokers' daily diet is significant lower than in non-smokers. Smokers consumed significantly higher saturated fatty acids comparing to non smokers. In conclusion, smokers have unhealthy diet and it may intensify harmful effects of smoking in their organism. PMID- 18409282 TI - [The effect of tobacco smoking during pregnancy on concentration of uric acid in matched-maternal cord pairs]. AB - Intensity of oxidative stress caused by prooxidant-antioxidant imbalance may have a pathophysiological effect in fetus of women smoking during pregnancy. Uric acid is the main contributor (60%) to TRAP (total radical trapping antioxidant parameters) which is markers of the plasma total antioxidants capacity. Therefore the aim of the study was to estimate the effect of tobacco smoking on concentration of uric acid in blood of pregnant women and in umbilical cord blood. Correlations between uric acid and total plasma antioxidant capacity in mothers and their babies were also studied. Healthy, pregnant women (n=147) were divided into groups non-smoking and smoking according to questionnaire declaration and concentration of cotinine in serum and urine. Concentration of uric acid in plasma was measured by enzymatic method using available kits from bioMERIEUX (France). We observed that, in both of studied group plasma concentration of uric acid increased during pregnancy. In group of smoking women level of uric acid was lower by 20% in I, by 25% in II and by 15% in III trimester than in group of tobacco abstinent. The differences were statistically significant, although the range concentrations in individual patients in both studied groups were similar (from 102.0 micromol/l to 408.0 micromol/l). In umbilical cord blood of children of smoking mothers mean level of uric acid was lover by 18% than in control group. Plasma concentration of uric acid was significantly correlated with total plasma antioxidant capacity both in smoking (r=0.75; p<0.001) and tobacco abstinent (r=0.60; p<0.05) pregnant women. The similar correlation was observed in umbilical cord blood in both groups (smoking: r=0.74; p<0.001, non-smoking: r=0.60; p<0.05). Reduced concentration of uric acid in plasma of women smoking tobacco during pregnancy suggest, that utilization of this antioxidant for neutralization of free radical present in cigarette smoking is enhanced and may have a negative effect on total plasma antioxidant capacity in mothers and their babies. PMID- 18409283 TI - [The concentration of the pregnancy-associated plasma protein A (PAPP-A) in pregnant women smoking tobacco--preliminary study]. AB - An improvement in prenatal screening for chromosomal defects in the first trimester of pregnancy has been achieved by combining sonography and biochemical markers. Among these markers exist the metalloproteinase pregnancy-associated plasma protein-A (PAPP-A) which promotes cell growth by proteolytic cleavage of insulin-like growth factor binding proteins (IGFBPs) causing the release of IGFs. The reduced level of PAPP-A in smokers in first-trimester has been reported previously. However, smoking influence on maternal level of PAPP-A during pregnancy has not been completely determined. The aim of this preliminary study was the investigation of PAPP-A concentrations in smoking women in the first- and third-trimester of pregnancy. The study comprised 40 pregnant women: 20 nonsmokers and 20 cigarette smokers. Only normal singleton pregnancies were included. Maternal serum levels of PAPP-A were measured with the KRYPTOR rapid random-access immunoassay analyser (Brahms GmbH, Germany) using time-resolved amplified cryptate emission technology (TRACE). In first-trimester of pregnancy we observed the lower of PAPP-A median values in smokers (2.77 IU/L, range 1.12 9.28 IU/L) by 24% than in nonsmokers (3.64 IU/L, range 1.68-6.79 IU/L). Next, we indicated that in third-trimester of pregnancy the PAPP-A median values were lower in smokers (48.45 IU/L, range 15.18-176.2 IU/L) until by 50% in comparison to the controls (96.56 IU/L, range 25.59-164.0 IU/L). Further studies will be continued to assessment the influence of some components from tobacco smoke on maternal serum PAPP-A in all trimesters of pregnancy. PMID- 18409284 TI - [Serum folate and homocysteine concentrations in women smoking during pregnancy and in umbilical cord blood of newborns]. AB - In metabolism of homocysteine several enzymes and vitamin cofactors are involves. Genetic abnormalities in these enzymes or nutritional deficiency vitamins, especially of folate may lead to hyperhomocysteinemia, a known risk factor for some pregnancy complications. High maternal homocysteine and low folate levels correlate with low birth weight. Maternal smoking affected significantly total homocysteine concentration in infants. Studies in this area are still scarce and report on limited number of patients. The aim of our study was to assess serum folate and total homocysteine (tHcy) concentrations in smoking pregnant women and in their newborn infants as compared with nonsmoking. The study consisted of 57 pregnant women, who qualified into two groups: smoking (n=28) and nonsmoking (n=29). The serum concentrations of folate were determined by electrochemiluninescent method and tHcy by fluorescence polarization immunoassay. We shown, that serum homocysteine concentrations were significantly higher in smoking as compared with nonsmoking pregnant women (p<0.05) as well as in umbilical cord blood of their newborns (p<0.001). The folate levels were comparable in serum both groups of mothers, but in infants born to smoking women were lower by 20%. In addition, the maternal serum levels of homocysteine and folate showed a significant positive correlation's with these parameters in newborns. Average birth weight infants born to smoking mother was significantly lower than nonsmoking cigarettes (p<0.05). It seems that tobacco smoking during pregnancy affected folate and homocysteine levels in serum of mothers and their infants. Smoking exposure is also associate with reduced birth weight. PMID- 18409285 TI - [The influence of tobacco smoke on homocysteine level in plasma of healthy males]. AB - The aim of the study was to evaluate the influence of tobacco smoke on homocysteine concentration in plasma. 168 healthy males between the age of 18 and 60 were qualified for the study. The exposure to tobacco smoke in examined group was determined on the basis of personal questionnaire and cotinine concentration in plasma. Three groups were chosen: non-smokers (control, 53 people), passive (48 people) and active smokers (67 people). The average levels of plasma cotinine concentration for selected groups were 4.3; 18.3 and 115 ng/ml respectively. Significant difference (p=0.019) of plasma homocysteine concentration between passive smokers and non-smokers was determined (10.4 and 9.2 micromol/L, respectively). A difference of homocysteine concentration in plasma for active smokers relatively to control group was higher and was 12.2 vs 9.2 micromol/L, p<0.001. Concentrations of plasma homocysteine were significant and positively correlated with concentrations of cotinine in plasma and Pearson's correlation coefficient (r) was 0.341 for passive smokers and 0.678 for active ones. The results obtained indicate that passive smoking has effect as toxic as active smoking which, inter alia, is manifested by increase in concentration of aterogenic homocysteine in plasma for both passive and active smokers. PMID- 18409286 TI - [Smoking influence on the level of homocysteine and 5-methyltetrahydrofolic acid in active and non smokers]. AB - Cigarette smoking is associated with oxidative stress and increased risk of many chronic diseased. Smoking inducts depletion of cellular antioxidant and is also known to be associated with an increased homocysteine level. Exposure to tobacco smoke has negative impact on the folic acid level. Folic acid is cofactor by demethyla-tion of homocysteine to non toxic methionine. The aim of this study was to determine the concentration of total plasma homocysteine and 5 methyltetrahydrofolic acid in active and non smokers and to evaluate the influence of the tobacco smoke exposure on urinary cotinine levels. The results indicated significant increase of total plasma homocysteine in smokers, than non smokers. We also investigated the negative influence of tobacco smoke on the level of folic acid in plasma and it suggests, that additional supplementation of folic acid in smokers is necessary. PMID- 18409287 TI - [Cigarette smoking results in the number of CD8+Fas Ligand+ T cytotoxic lymphocytes in bronchoalveolar lavage (BAL) fluid of patients with idiopathic pulmonary fibrosis (IPF)]. AB - Idiopathic pulmonary fibrosis (IPF) is an interstitial lung disease with unfavourable outcome. Tobacco consumption in IPF exacerbates the clinical manifestations and limits the time of patient survival. The cyto-immunological alterations caused by smoking in IPF patients need particular explanation. BAL was carried out in 21 non-treated patients with IPF, subdivided according to the smoking status (n=7 for smokers). BAL routine cytology was completed by; immunotyping, including T cell major subsets (CD4 and CD8) stained for Fas, Fas ligand (FasL) and TNFR-1, late apoptosis/cell cycle analyses (BAL cells were permeabilized and stained with PI) and TUNEL assay. BAL cytology in IPF, as compared with control group, was characterized by significantly higher total cell and macrophage number, increased lymphocyte, neutrophile and eosinophile percentage and relatively low CD4/CD8 ratio. Cigarette smoking in IPF resulted in enhanced BAL lymphocyte CD8 cell percentage and number, as compared with nonsmoking subgroup and further decline in CD4/CD8 ratio (0.41+/-0.15 vs 1.23+/ 0.29 in nonsmokers, median +/- SEM, p<0.05). The percentage of CD8, but not CD4 cells carrying Fas Ligand was significantly increased in IPF smokers (12.0+/-3.1% vs 3.7+/-0.9% in nonsmokers, median +/- SEM, p<0.05). Apoptosis rate of BAL macrophages and lymphocytes was enhanced in IPF, as compared with controls (confirmed by both techniques), but without remarkable changes, if compared one IPF subgroup to another. The number and percentage of CD8+FasL+ was negatively correlated with vital capacity (VC) values in IPF patients, but not with BAL inflammatory cell apoptosis rate. Cigarette smoking enhanced a percentage as well as a total number of both BAL CD8 and BAL CD8+FasL+ cells in IPF patients. BAL cytotoxic cells (CD8+FasL+ lymphocytes) seem to have impact on impaired lung function in smoking IPF patients. PMID- 18409288 TI - [The evaluation of level of epigenetic indicator--5-methylcytosine in smoking and non-smoking patients with mild essential hypertension]. AB - 5-methylcytosine (m5C, 5mC) is a nucleotide occurring naturally in genomic DNA and play an important role in regulation of genes expression. Methylation of cytosine in DNA is an epigenetic modification and different intrinsic and extrinsic factors can influence on its level. For example, it is subject to modification and/or degradation by the free radicals which are commonly present in environment of human, among others the cigarette smoke. The reactions of m5C with free radicals lead to origination of many products which effect is decrease of level of m5C in DNA (hypomethylation) and excessive expression of genes inducing development of different diseases, especially cardiovascular system diseases. The aim of the study was statement if exist differences of level of 5 methylcytosine in DNA between smoking and non-smoking patients suffering from mild essential hypertension. The study group was composed of 30 patients suffering from mild essential hypertension (21 females and 40 males) aged from 18 to 55 years (32.4+/-10.3 years). The group of smoker was composed of 13 patients (5 females and 8 males) and the group of non-smokers was composed of 17 patients (7 females and 10 males). 3-5 ml of blood was sampled on EDTA and then thin-layer chromatography analysis of 5-methylcytosine level in DNA after previous enzymatic hydrolysis of DNA and radioactive phosphorus labeling [32p] was performed. The mean level of 5-methylcytosine (m5C) were 1.30+/-0.56 [%] in non-smoking patients, and 1.28+/-0.42 [%] in smoking patients suffering from mild essential hypertension. There is no significant statistically differences between non smoking and smoking patients (p>0.4). In the study the following conclusion was drawn: the level of m5C in DNA of patients suffering from mild essential hypertension in the study is independent of smoking (p>0.4) in patients with mild essential hypertension. However it supposes, out of regard for theoretic datum suggestive such influence, the study should be performed in more frequent group of patients. PMID- 18409289 TI - [Does tobacco smoke influence the concentration of asymmetric dimethylarginine (ADMA) in healthy males plasma?]. AB - Toxic effects of tobacco smoke may manifest by the increase of concentration of aterogenic compounds in the human body. One of them is asymmetric dimetyloarginine (ADMA) which is perceived not only as a significant risk factor of endothelium dysfunction and cardiovascular disease but also predicts all-cause and cardiovascular mortality independently of established and emerging cardiovascular risk factor. The aim of this work was to check to what degree ADMA concentration in plasma changes for people exposed to tobacco smoke in various intensity. The study population consisted of 168 healthy males between the age of 18 and 60. The exposure to tobacco smoke in examined groups was determined on the basis of questionnaires and cotinine concentration in plasma. Three groups were selected: non-smokers (53 people), passive smokers (48 people) and active smokers (67 people). Insignificant differences of ADMA concentration between passive and active smokers in relation to non-smokers were determined, 15.4% (0.45 vs. 0.39 mmol/l,) and 7.7% (0.42 vs. 0.39 mmol/l) respectively. There was no significant relationship between concentration of ADMA and cotinine in plasma (r=0.173; p=0.239 for passive smokers and r=0.218; p=0.076 for active smokers). The results obtained indicate that tobacco smoke has small influence on ADMA level in healthy males' plasma. PMID- 18409290 TI - [Is hypotensive effectiveness of amlodipine therapy in smokers connected with endothelin-1 (ET-1) concentration?]. AB - An interaction between a vasoconstrictive factor--endothelin-1 (ET-1) and nicotine is currently a subject of investigations. Many studies make us consider whether worse hypotensive effectiveness of many antihypertensive agents applied in smokers may be connected with elevated ET-1 level. The aim of this study was evaluation of hypotensive effectiveness of amlodipine in smoking (group A) and non-smoking (group B) hypertensive subjects. Assessment of plasma ET-1 concentration in group A and B and evaluation of influence of 30-day amlodipine therapy on plasma ET-1 concentration in group A and B. The examined group comprised 31 patients with essential hypertension (grade I and II). The result of the amlodipine therapy (5 mg/day) was statistically significant decrease in systolic (SBP) and diastolic (DBP) blood pressure in group B. In group A non significant reduction of SBP and DBP was observed. In group A plasma ET-1 concentration was significantly higher than in group B. We noticed non significant decrease in plasma ET-1 concentration in both groups. These results do not prove the hypothesis that worse hypotensive effectiveness of amlodipine applied in smokers may be connected with elevated ET-1 level. Despite of higher ET-1 concentrations in hypertensive smokers one could not observe significant influence of the therapy on changes of plasma ET-1 concentration in both groups. PMID- 18409291 TI - [Cervical cytology picture--tobacco smoking and inadequate patients' history taking]. AB - Cardio-vascular diseases as well as neoplasms (especially upper respiratory cancer and lung cancer) the most commonly mentioned consequences of tobacco smoking. But among tobacco-related diseases there are several other conditions including cervical cancer. In the present study we included 1166 females who underwent gynecological cytology examination at Obstetrical and Gynecological Office in University Hospital in Bydgoszcz from January till June 2007. All samples were stained according to Papanicolaou procedure and evaluated according to Bethesda classification. We analyzed also information regarding smoking habit which should be included in patients' form. In our studies we found that in 67% of cases physicians did not indicate patients' smoking status. According to patients records we received only 3% were smokers while 30% were non-smokers. In cytological smears from smokers we found about 3 times higher incidence of high grade lesions comparing with non-smokers (6% vs. 1.7%). However, atrophic and inflammatory changes were more often fund among non-smokers (30% vs. 18%). In conclusion, we can state, that in almost 70% of studied cases smoking habit information was ignored by clinicians. Lesions with high risk of cervical cancer were found more commonly in smoking patients. PMID- 18409292 TI - [Influence of smoking on selected microelements in serum and hair of adults]. AB - The aim of this study was to assess the concentrations of iron, zinc and copper in serum and hair of smokers and nonsmokers. The study population comprised 162 subjects aged 30-60 years, in this 47 smokers and 115 non smokers. All subjects had their blood and hair samples collected for analysis. Content of the analysed microelements was assayed with atomic spectrophotometry using Zeiss AAS-3 spectrophotometer. Statistical analysis was done with Statistica 6.0. The results showed that smokers had significantly lower concentration of copper in serum than non smokers. The analysis of molar ratio of minerals in serum showed significantly lower ratio of zinc to copper in smokers as compared to non smokers. The content of minerals in hair of both groups was insignificant. Based on the obtained results it is justified to conclude that smoking can influence on copper metabolism in organism. PMID- 18409293 TI - [Influence of passive smoking on content of aluminium in pharyngeal tonsils from children living in Southern Poland]. AB - The characteristic of occurrence Al content in pharyngeal tonsils from girls and boys living in Southern Poland is presented in this article. Analysis of content of Al in pharyngeal tonsils was observed in four groups of children: girls exposed to tobacco smoke and unexposed to tobacco smoke, boys exposed to tobacco smoke and unexposed to tobacco smoke. Arithmetic mean of contents of aluminium in pharyngeal tonsils from exposed girls was 15.94+/-8.08 microg/g, in unexposed girls was 14.11+/-5.08 microg/g, in exposed to tobacco smoke boys--16.68+/-11.17 microg/g and unexposed boys 18.08+/-10.56 microg/g was observed. Differences between analysed groups of children were not statistically important. The cross correlation analysis between content of Al and other metals Ba, Ca, Cr, Cu, Fe, Mg, Mn, Zn, K, Na showed repeating co-dependences between Al and Fe, Mg and Zn in pharyngeal tonsils from children exposed to tobacco smoke. In unexposed children was only one co-dependence between Al and Ca. PMID- 18409294 TI - [Influence of environmental tobacco smoking on different barium contents in pharyngeal tonsils]. AB - The aim of this study was analysis of content of barium in pharyngeal tonsils from children living on Southern Poland in during exposure on ETS. Barium contents in pharyngeal tonsils were higher for no exposure ETS children (0.099 microg/g) in comparison to exposure by ETS (0.033 microg/g), p=0.0042. The interaction between Ba and Ca, Fe, K or higher arithmetic and geometric mean, statistical range of changes contributed the role ETS in relation to children. PMID- 18409295 TI - [Influence of smoking tobacco on the occurrence metals in some parts and profiles of femur head]. AB - The smoking tobacco makes the negative influence on state of osseous tissue, many research shows, that the smoking tobacco states the factor of risk of appearance of osteoporosis. The aim of this study was determined of occurrence of 12 metals in separate parts and profiles of femur head in three studied groups: smoker persons, non-smoker and smoker in past. Human femur heads were obtained from people living in Upper Silesia cities in Poland during endoprosthetoplasty of coxa. The femur heads were cut into 5 cross-sections A-E using a stabilizing device provided with a fixed and a movable part maintained at a 12 degrees angle. As the next step, fragments of articular cartilage, cortical bone and trabecular bone were sectioned off from the cross-sections. In smoker group the large content of chrome of, nickel, zinc pays back attention. In case of individual parts of femur head notice, that the largest place in articular cartilage has accumulating metals. The largest contents of metals were observed in the external sections A and E. PMID- 18409296 TI - [Lead content in blood of children living near zinc smelter plant exposure on environmental tobacco smoking (ETS)]. AB - In this paper the results of concentration of lead in the blood of children living near and background the plant smelter are presented. 510 girls and 567 boys from schools located at Tarnowskie Gory and Miasteczko Slaskie took part in the study. Concentration of lead content in the blood of children was determined by the ASA method with the apparatus Pye Unicam SP-9. Smoking of parents increased the level of lead in the blood. Higher concentration of lead was show in blood girls (8.09 microg/100 ml) and boys (9.78 microg/100 ml) exposure to tobacco smoke in comparison to girls (6.19 microg/100 ml) and boys (6.81 microg/100 ml) living in no smoking conditions. PMID- 18409297 TI - [Nicotine in the hair as a biomarker of tobacco smoke by pregnant women- preliminary study]. AB - Determination of xenobiotics in hair has become common in evaluation of exposure to drugs. It can be used to assess the drug intake among adults and exposure to these substances in prenatal period. The aim of this study was the nicotine determination in hair of newborns' whose mothers were smoking during pregnancy. Also the relationship between concentration of cotinine--the main metabolite of nicotine in the urine of mothers and nicotine concentration in their infants' hair has been evaluated. The group of 15 women who had been smoking during pregnancy and 10 non-smoking and not exposed to ETS women and their newborns were the subject of the study. The cotinine levels in maternal urine samples was measured by the high performance liquid chromatography (HPLC) with a spectrometric detection, with the use of norephedryne as an internal standard, after a prior extraction with the use of liquid-liquid technique. Nicotine assess in infants' hair was carried out by means of gas chromatography coupled with mass spectrometry, with the use of ketamine as an interior standard, after the prior extraction with the use of liquid-liquid technique. The mean cotinine concentration in the group of smoking women amounted to 1132.5+/-1236.3 ng/mg creatinine and the level of nicotine in hair of children amounted to 1.9+/-3.2 ng/mg of hair. In the group of non-smoking women the urine cotinine test showed that two patients were exposed to the tobacco smoke. Nicotine in their children's hair was below the detection limit. The demonstrated correlation between the nicotine concentration in infants' hair and the cotinine levels in mothers' urine shows that the source of fetal exposure to tobacco smoke constituents is smoking during pregnancy. The applied gas chromatography method/mass spectrometry method allows to evaluate the level of nicotine in hair of the newborns, whose mothers were smoking during pregnancy, however it seems that in the case of exposure to ETS a more sensitive method, such as gas or liquid chromatography coupled with tandem mass spectrometry has to be applied. PMID- 18409298 TI - [Application of the immunoassay for determination of cotinine in the urine]. AB - Verification of the results of questionnaire studies of exposure to tobacco smoke require determination of the biomarkers in body fluids. For performing such measurements the fast and simple method is needed. The aim of this study was to estimate usefulness of enzyme-linked immunosorbent assay (ELISA) in routine cotinine determination in urine of active smokers and compare it with reference high performance liquid chromatography method (HPLC). In the study participated 15 non smoking and 15 smoking women. In the urine of tese women cotinine concentration of cotinine was measured by the means of HPLC and ELISA methods. The ELISA method, used in quantity measurement of cotinine in urine, is of great accuracy, sensitivity and specificity which were proved by comparison with high performance liquid chromatography (HPLC) reference method. Number of false positive and false negative results obtained by ELISA method did not exceed 10%. A high correlation coefficient, r=0.9056, between results of determining cotinine in urine by means of ELISA and HPLC methods, confirmed the utility of ELISA method to estimate the tobacco smoke exposure. The differences in cotinine concentration values obtained by ELISA and HPLC methods did not depend on cotinine concentrations in the urine samples. Although numerous advantages of ELISA method used to detect and determine cotinine, it should be taken into consideration that results might be overestimated and cross-reactivity with other xenobiotics present in urine must be concern. PMID- 18409299 TI - [Concentration of cotinine and trans-3'-hydroxycotinine and its glucuronides in urine of smoking pregnant women]. AB - To assess the exposure to tobacco smoke constituents, the biomarkers are used. In most studies conducted among active smokers, the cotinine is usually utilised as such a biomarker. The aim of this study was to estimate an application of determining two nicotine metabolites, cotinine and trans-3'-hydroxycotinine (3 HC) in order to assess cigarette smoking among pregnant women. There were 25 patients (15 smokers, 10 non-smokers) admitted to Delivery Ward in Gynecology and Obstetrics Clinical Hospital of the University of Medical Sciences in Poznan. The free and total form of cotinine and trans-3'-hydroxycotinine in urine were determined by the means of high performance liquid chromatography with spectrometry detection. The results of this studies indicated that in urine both cotinine and trans-3'-hydroxycotinine are conjugated with glucoronide acid in the high degree. The linear correlation between free and conjugated form of cotinine and whole cotinine concentration indicates the possibility of application of all three of them as biomarkers of tobacco smoke exposure. The high conjugation of trans-3'-hydroxycotinine with glucuronic acid (over 80 %), and high correlation between glucuronide and total trans-3'-hydroxycotinine concentration proves necessity of having the urine samples hydrolysed or determining the 3' hydroxycotinine glucoronide, when application of 3-HC as a biomarker of tobacco smoke exposure is concerned. Our studies confirm other authors' observations that division of 3'-hydroxycotinine and cotinine concentrations might be used as a predictor of differences in cotinine metabolite ratio (polimorphism). The studies shown that using analytical equipment, nowadays available in the most laboratories (HPLC), it is possible to determined two biomarkers which are very useful and give various information, no matter whether they are conjugated with glucuronic acid, or not. PMID- 18409300 TI - [Selected psychosocial factors in 15-year-old current smokers and ex-smokers]. AB - The aim of the paper was the study selected differences between 15-year-old adolescent current smokers and ex-smokers with regard to: self-efficacy, family relations, school psychosocial environment and perception of peer smoking. The anonymous survey was carried out in 2006, within HBSC study (Health Behaviour in School-aged Children), using a representative sample of 2287 persons in the mean age 15.7 years. Optional HBSC 2001/02 question about smoking status was applied again in Poland (i.e. whether a student is a smoker, ex-smoker or never smoker). We compared a group of N=365 smokers and N=384 ex-smokers, while never smokers were temporally excluded from our analysis. The international standard questionnaire was used. The results showed that adolescent ex-smokers in comparison with current smokers had more often higher self-efficacy (41% and 35.3%), assessed family relations as very good (42.1% and 31.4%), more seldom assessed school psychosocial environment as poor (40.5% and 52.1%), and more seldom considered their friends as heavy smokers (38.3% and 67.9%). The differences between smokers and ex-smokers were statistically significant mostly in girls. In the prevention strategies it is very important to take under consideration gender-specific psychosocial characteristics including: individual factors, family, school and peer environment. Our findings reveal that many psychosocial factors may be protective against tobacco dependence. PMID- 18409301 TI - [Smoking among school children from the Wielkopolska region]. AB - Smoking defines as a gate open for development other addictions in a group of adolescents. The aim of the study was the estimation the phenomena of smoking among children and adolescents as well as the knowledge concerning the negative effects of it in Big Poland area in every type of school. In the study participated 1533 pupils from every kind of schools. The questionnaire own concept was used in the study. It estimated the frequency of smoking and knowledge concerning the effects of smoking in respondents. The results underwent the statistical analysis. The results show that 88.78% children and adolescents state that they do not have any problems with smoking. 37% non-smoking pupils state they started smoking in the past. The most important reason for smoking was the influence of social environment - at school or in a place of living. The age structure shows the pupils start to smoke between 13-15 yrs of age. 56.4%. The knowledge about negative effects of smoking is on media level, in most case true. Pupils try to stop smoking often but the trials are not as effective as they expect. The result show there is the necessity of making some efforts toward more effective educational activities and preparation some specialist support in addiction treatment. PMID- 18409302 TI - [The analysis of smoking conditionings for junior high school students]. AB - The use of psychoactive substances by children and teenagers is an important public healthcare issue in Poland. Starting smoking by teenagers increases the likelihood of addiction to smoking later as well as getting numerous smoking related diseases. The aim of the study was to examine the smoking prevalence and its conditionings for the 15-year-olds living in a city of 60.5 thousand people as well as provide the group profile. The study included 114 randomly selected students. 9.6% of the study group were smokers, out of whom 3.5% junior high students were occasional smokers and 6,1% declared everyday smoking (boys only). Over half of the students (57.1%) have never smoked. There has been a coexistence of smoking and using other psychoactive substances. The smokers used vodka or other types of alcohol far more often than the non-smokers, and they become drank. All of them have been drunk at least once. Approximately 40% of the smokers experimented with drugs. The conditionings of smoking for junior high school students have been defined. The study showed an important relation between smoking by junior high school students and being raised in dysfunctional family or being raised in a healthy lifestyle family of non-smokers. PMID- 18409303 TI - [Cigarette smoking by secondary school students in the country of Rzeszow]. AB - A last ten years cigarette smoking has become very popular. The most startling finding is the increase in the amount of young people who are becoming smokers. At is well known, that smoking cigarettes causes irreversible changes in the bodies of young people. The main objective of the thesis was to document and interpret the spread of cigarette smoking among secondary school of aged students in Rzeszow country. The research was carried--out in June, 2007. 193 students took part in the research study, they were between the ages of 13 and 16 years old, 52.84% were girls and 47.16% were boys. This group of young people was specifically selected to cover both the city and its surrounding area in the country of Rzeszow. Of the students who took part in the research 103 (53.36%) were from the surrounding countryside and 90 (46.63%) from the city. The research findings were gathered using diagnostic techniques, including questionnaires prepared by the author. To analyze the statistics the author used the test of independence chi Parsons Square, and the variety analyze (ANOVA). The research showed that the majority of young people (83.94%) withheld the fact that they were smoking cigarettes. 67.24% of the students in the study were secondary school aged boys. Most of them (83.87%) smoke between 1 to 5 cigarettes per day. Even for those who smoked, when asked they said, that health was most important to them. It is surprising, that smokers who smoke everyday chose health as their most important value. After analyzing the conditions which affect smoking, we found that many of their colleagues (77.20%), teachers (63.73%) and parents (39.89%) smoked also; although that didn't affect the regularity of smoking by the students surveyed. PMID- 18409304 TI - [Smoking in the aspect of wholesome behaviors in adolescents]. AB - It has been compared behaviour of wholesome adolescents (14-17 years old) in this study, the healthy and suffering from epilepsy and headache. The IZZ and LKZ research methods have been used. The research group has been divided according to sex and age. It has been proved the greater escalation of health-oriented behaviour in patients, particularly girls. PMID- 18409305 TI - [The problem of tobacco addiction among high-school teenagers]. AB - Tobacco addiction is a serious health and economic problem concerning the whole globe. High-school teenagers are a group of society whose habits, knowledge about harmfulness of smoking tobacco have an influence on future development. The aim of the thesis was to define the problem of high-school teenagers' addiction to nicotine. The research was conducted among 341 young people from different high schools. The research group consisted of students: High School no. 11 and, Artistic High School in Poznan, High School in Klodawa and two schools in Norway, Askim Ungdomskole--teenager school and Videregaende Skole in Askim--high school. The representative sample group numbered: 287 in Poland and 54 persons in Norway. Smoking cigarettes and addiction to nicotine is considered to be a medical problem, which is related to the occurrence of many diseases, as well as a social problem which regards the biggest part of youths. PMID- 18409306 TI - [Pharmacy students' exposition to tobacco smoke--a trial of assessment of causes and prevalence]. AB - Prevalence of tobacco smoking among young people is one of the most important aspects in prevention of smoking and smoking-related diseases. The aim of this study was assessment of pharmacy students' exposition to tobacco smoke (active and passive smoking) and searching for circumstances promoting tobacco smoking and exposition to ETS (environmental tobacco smoke). In the examined cohort of 161 students we found 37 smokers (23%; 21.2% women and 31% men), among them 14 (8.7%) everyday smokers. ETS affected 155 persons (96.3%) and usually took place in a circle of friends. The most common factors promoting tobacco smoking were: pleasure connected with smoking, smoking as a support in stress situation or as a facilitation of society functioning and getting to know new friends. We must emphasize again the leading role of prevention of smoking initiation in young people in all preventive activities. PMID- 18409307 TI - [Students of the Faculty of Medicine and of the Division of Public Health of Medical University of Bialystok towards nicotine addiction against the background of the National Health Program in Poland]. AB - In the years 1998-2005 in Poland a National Health Program was being implemented. One of its chief guidelines was to reduce the popularity of tobacco smoking, recognised as a proven, single factor influencing the etiology and the course of many diseases. The objective of the work was to establish whether the program's guidelines were reflected in individual attitudes of senior students of the Faculty of Medicine and of the Division of Public Health of Medical University of Bialystok. The research was conducted with the use of an anonymous questionnaire, filled in by the students before the classes dedicated to the issues of nicotine addiction within the framework of the family medicine thematic block. The analysis of the results did not show a decrease in the number of student smokers in comparison to the previous years; however, it was observed that more than a half of the current smokers had tried and was going to try to give up smoking, and the non-smokers strongly objected to passive smoking. PMID- 18409308 TI - [The phenomenon of spreading smoking cigarettes among the students in Institute of Cosmetology and Heath Care in Bialystok]. AB - Tobacco smoking is one of the principal public Heath problems. The Word Heath Organization declared the addiction of tobacco smoking as a separate disease, International Classification of Diseases, ICD number F17. The health service workers play a huge part in establishing pro-health attitudes. They are expected to participate in campaigns promoting a tobacco-free lifestyle. It is important that the persons mentioned were nonsmokers. The main goal of the study was to estimate the phenomenon of spreading smoking cigarettes among the students and accomplishing their knowledge and awareness about threats resulting from tobacco smoking. The study was performed among 501 students. They answered questions concerning smoking and knowledge about nicotine dependence. The research touched such questions as conviction about damage caused by smoking, the source of information, causes and effects of smoking, about environment whether they are, the interviewee's smoking preventive activities. PMID- 18409309 TI - [Frequency of smoking tobacco among the students of the last year of the Faculty of Health Sciences]. AB - Smoking tobacco is one of the most frequent and most dangerous addictions among the Poles, at the same time--despite the many dramatic results--it is the most belittled of threats. It is difficult to understand especially those smokers who, due to their future or present job should be free from tobacco smoke. The aim of the work was to establish the participation of the smoke inhalers among the students of the last years of studies, focusing on the particular socio demographic features. 162 students were tested, that means all who are the last year students at the Faculty of Health Sciences of the Medical University of Lodz. Using the auditoria survey, the studies were carried out between the 1st to 15th March 2007. The filled in surveys were handed back in by 92.6% of students (150 female and male students). Among the 150 of the tested, 58 people confessed to smoking (38.6%). The ratio of the smoking female students was 34.0% and smoking male students 46.4%. In the past, there were close to 65% of smokers among the tested. Over 54% of the asked people smoked their first cigarette in the high school. Majority of smokers (30.5%) smoked from 5 to 10 cigarettes a day. Majority of smokers (70.4%) confirmed they smoked everywhere where they wished. From among 58 smokers, 4 people could be pharmacologically addicted to nicotine. Almost all of them would like to quit smoking. The ratio of smoking students of the last years of the Faculty of Health Sciences of the Medical University of Lodz was very high in 2007. There was practically every second male student who smoked and close to every third female one. Great majority of the smokers put the health of the people around them who did not smoke at risk because they smoked everywhere they pleased. There is a need to undertake some efficient preventive actions directed at the problem of smoking among the students, especially of the departments which produce the personnel of the health centres. PMID- 18409310 TI - [Tobacco smoking among students from Universities in Rzeszow]. AB - The habit of tobacco smoking has existed for centuries, it has spread all over the world and become one of the most important health risk factors. It is assumed that 21 illnesses are related to tobacco smoking. Among those illnesses there are the following: tuberculosis, some types of cancer, circulatory system diseases, pneumothorax, chronic obliterative pneumonia, bronchitis, flu, stomach ulcer and some diseases among babies such as low birth weight and the syndrome of sudden death. Systematic monitoring of smoking cigarettes among students who are especially prone to smoking addiction can be used in activities concerning so called health risk management. The aim of the study was to try to assess tobacco smoking among students from the city of Rzeszow representing different faculties. 555 students representing two largest Universities in Podkarpackie Province: Rzeszowski University and Technical University were asseesed in this study. To present the data statistically the chi-square test of independence was used. The studies show that most of the students have never smoked cigarettes (71.2%) and more that a half of the students who smoke regularly say that they might give up smoking in the future. The number of cigarettes smoked daily was different on different faculties. Students representing humanities and science faculties smoke more cigarettes daily than students representing medical faculties. PMID- 18409311 TI - [Tobacco smoking amongst students in the Medical Faculty of Wroclaw Medical University]. AB - The main aim of health promotion and diseases profilaxis is a struggle with smoking, which is a well known factor in many disorders, i.e. malignant carcinomas, noncarcinomatous diseases of respiratory system and cardiovascular diseases. The aim of the study was the analysis of the smoking level amongst the students of 6th year of the Medical Faculty of Wroclaw Medical University. Amongst 131 polled women--116 were non-smoking persons (88.5%), 15 smoking (11.5%). Amongst 55 polled men--43 these are non-smoking persons (78%), 12 smoking (22%). Out of smoking women, the most women (6) is smoking from 11 to 15 cigarettes per day, out of smoking men, the most (5 men)--6-10 cigarettes per day. Smoking women began smoking during the secondary school the most often. 8 of men began the smoking in the secondary school. The majority of examined didn't try to limit smoking. Respondents would expect the biggest support from close persons during giving up smoking Only 59% of women and 64% of men disagree definitely to smoking in their presence. In the consequence of above results surprising seems still high percentage of smokers amongst examined, scantiness of taking attempts of giving up smoking and indifference of non-smoking medical students towards smoking in their presence. PMID- 18409312 TI - [The knowledge level of smoking harm in the students of Wroclaw Medical University, Bialystok Medical University and the Public Higher Medical Professional School in Opole]. AB - The main aim of health promotion and diseases prophylactic is a struggle with smoking, which is a well known factor in many disorders, i.e. malignant carcinomas, noncarcinomatous diseases of respiratory system and cardiovascular diseases. The aim of the study was the analysis of the knowledge level of smoking harm and its consequences in 1051 students of Wroclaw Medical University, Bialystok Medical University and the Public Higher Medical Professional School in Opole. The respondents answered to the anonymous, voluntary questionnaire. The little percentage of students in all centres gave correct answer to the subject of amount of carcinogens contained in the tobacco and the tobacco smoke still correct answers concerning concrete carcinogens were rare. Students aren't also convinced that the smoking can cause so strong psychophysical addiction, like taking drugs: heroine and cocaine. Majority polled is confirming that the smoking is a cause of the cancer of larynx, vascular diseases or chronic bronchitis, as well as an influence on a birth weight in newborn babies. Depending on the examined centre--convincing that smoking is the risk factor of bladder cancer isn't already so universal, the similar situation is taking place at examining the knowledge on the subject of association between smoking and osteoporosis. Admittedly the knowledge on the subject of nicotine substitute therapy isn't alien to students, they have the difficulty with correct giving available preparations on the Polish market. The knowledge of students is also scarce on the subject of changes in the total number of smokers in Poland. PMID- 18409313 TI - [Comparison of the knowledge level of stationary and non stationary students of the Public Higher Medical Professional School in Opole on the subject of the health results of smoking]. AB - The smoking is a multifaceted social problem. Early beginning smoking by children and young people are increasing the risk of the quicker appearance of tobacco related diseases and mortality of people in a productive age. It is a cause of the higher death rate than AIDS, the alcoholism, car accidents, drugs, fires, homicides and suicides assessed together in the world. Even though smoking is making greatest risk, its prevention is possible and should be an element of the health education understood widely. The aim of the study was the ccomparison of the knowledge level of stationary and non-stationary students of the Public Higher Medical Professional School in Opole on the subject of the health results of smoking. Students of non-stationary studies demonstrated the higher level of the knowledge on the subject of health results of smoking. The stationary students, young people, don't realize a nicotine addiction as the strong risk fully--11.2% of examined is negating a fact that the smoking can cause so strong addiction, like taking drugs. PMID- 18409314 TI - [The analysis of smoking amongst students in the Public Higher Medical Professional School in Opole]. AB - Making conditional on the smoking is making the essential problem of the contemporary society. Conducted examinations are pointing in Poland and in the world that the number of young people which are reaching for the first cigarette is soaring together with age. The aim of the study was an analysis of smoking amongst students of the Public Higher Medical Professional School in Opole. Group of 290 stationary and non-stationary students of the Nursing Institute and the Obstetrics Institute of the Public Higher Medical Professional School in Opole was put through an examination. Out of polled --72 students (24.8%) is smoking cigarettes but 218 students (75.2%) are non-smoking persons. 13.8% of the examined group of non-stationary students is smoking cigarettes, however smoking cigarettes is declaring 29% of the examined on stationary studies. Polled the most, well as far as 70.8%, began smoking cigarettes during the secondary school. 62.5% smoking persons tried to give up smoking having the harmfulness of smoking in mind. Unfortunately these attempts ended in failure. The students would expect the biggest support from close persons (the husband, the wife) during giving up smoking, 93.1% of examined is confirming such an opinion. PMID- 18409315 TI - [Consumption of psychoactive substances--awareness of health consequences among the students of university schools in Poznan]. AB - The goal of this paper is to estimate the phenomenon of consuming psychoactive substances such as: alcohol, nicotine, caffeine and narcotics among the students of Poznan's universities, and evaluating the level of consciousness of the dangers resulting from using those substances. The authors wanted to check, whether the consumption of psychoactive substances depends on such traits as: sex, place of living, subjective evaluation of one's health, the type of university they attend, and whether the respondents think that the knowledge passed onto them on the universities about the dangers resulting from consuming such substances is sufficient, and whether they know how to help an addicted person. The research, done with the use of a survey, was conducted among 504 students from six universities in Poznan: Medical University (16.7% of the respondents), University School of Economics (15.3%), University School of Agriculture (162%), University School of Physical Education (16.1%), Poznan Technical University School (184%) and Poznan University (17.3%). The research has shown, that the most of the students consume alcohol (81.1% of the respondents), followed by caffeine (75.8%). The third place was taken by narcotics (38%), and the fourth by cigarettes (20%). Most people that smoke are the ones that are renting an apartment by themselves. No statistic difference was found in the usage of cigarettes between women and men, nor was there a relation between the subjectively evaluated state of one's health, or the attended university (the students of the Medical University smoke as much as the others). The average ago of the initiation into tobacco usage of the respondents was 17 years of age, which is a time when one doesn't have a legal right to obtain cigarettes. PMID- 18409316 TI - [Introduction to the Preventive Medicine Program "Prevention 2006-2009" in the Institute of Cosmetology and Health Care in Bialystok]. AB - Smoking is still a common habit in Poland. Nowadays, it is the way of coping with stress, it is used to become calm. It is also widely accepted by people from various backgrounds included medicine environment. Unfortunately, the number of young people--tobacco addicts is increasing. Survey proceeding in University implementing of the health promotion's program was to estimate the phenomenon of spreading smoking cigarettes among the students and accomplishing their knowledge and awareness about threats resulting from tobacco smoking. The study was performed among 501 students. They answered questions concerning smoking and knowledge about nicotine dependence. The research touched such questions as conviction about damage caused by smoking, the source of information, causes and effects of smoking, about environment whether they are, the interviewee's smoking preventive activities. Implanting health promotion's program is an interdisciplinary subject which glue together such sciences as medicine, psychology, sociology, social politics and many others. PMID- 18409317 TI - The influence of two generations of smokers on cigarette smoking among young people. AB - In the 21st century, 4.2 million out of 1.3 milliard cigarette smokers die every year. In the West Europe in the year 2002, 105.9 million people smoked cigarettes. In Europe the number of young smokers grows by 500 thousand every year. It is stated that 9 million active smokers live in Poland. Although generally the consumption of tobacco decreases, still the scale of the problem is large. In 1996 Polish government passed the health protection act concerning the consequences of smoking. Comprehensive actions bring measurable effects, because 77% of the Poles (among them 27% of smokers) want the prohibition of smoking in public places. The aim of the study was to show the influence of two generations of smokers on cigarette smoking among young people. 244 active smokers, who have smoked cigarettes consecutively, were recruited to the study. People recruited to the study were divided into two age groups. The first group consisted of people at the age from 18 to 25, the other of people at the age of 65 and older. The differences were statistically significant for young people between the age of 18 and 25 and for people > or =65 who had elementary (p<0.0002) and incomplete high education (p<0.000). In the young group twice more people (26.7%) started smoking in the primary school than in the senior group (p<0.009). Before matriculation exam there are 41.7% of regular smokers. It is twice more than the seniors when they were their age (p<0.008). In the other group girls up to 18 years of age smoked three times less often than boys. Their peers 40 years later (in the first group 41.7% of young people) are regular smokers twice more often. In the group of young smokers most of the participants smoked up to 5 cigarettes a day or half a package a day, when in the group of the participants aged 65 and older only 6.5% of them smoke up to 5 cigarettes a day (p<0.00002). Seniors prefer to smoke a package of cigarettes a day, some of them even up to 30 cigarettes per 24 hours (20.6%) which makes a significant statistical difference in comparison to the younger group (p<0.047). Older people smoke two packages daily four times more often than younger people. Only permanent anti-nicotine education of all social groups, cooperation with anti-nicotine clinics, non-government organizations, participation in campaigns fighting with nicotine dependence, may influence the further reduction of the number of smokers in Poland. Reminding parents about not smoking in the presence of children and sensitivity of the society to smoking adolescents may change the state of health of the Poles. PMID- 18409318 TI - [Exposure to tobacco smoke of pregnant women--results of prospective study in Lodz region]. AB - The aim of the study was to analyze the active smoking by pregnant women in different weeks of pregnancy. The study population consisted of 136 women before 22 weeks of pregnancy. The women were interviewed three times during pregnancy. From all women included into the study the saliva sample was collected three times to verified smoking status in pregnancy. Cotinine level in saliva was analyzes using Liquid Chromatography with Tandem Mass Spectrometry. Based on interview about 25-30% of pregnant women smoked cigarettes during pregnancy. This percentage was higher when the smoking status was verified using cotinine level in saliva (33-41%). In the group of pregnant women who smoked cigarettes there was no decreasing trend in cotinine level during pregnancy whereas in nonsmokers the decreasing trend was observed. Unmarried women, those with lover educational level, and unemployed had the highest risk of smoking during pregnancy. PMID- 18409319 TI - [Analysis of spreading of smoking habit among pregnant women admitted to the Prenatal Outpatient Clinic in 2005-2006]. AB - A harmful influence smoking on health is well known and documented. Smoking during pregnancy has negative influence not only on mothers but also on embryos. In spite of pregnancy is a very good reason to stop smoking only every third pregnant stop smoking during pregnancy. Nicotine, carbon monoxide and cyanide, components of smoke, are very toxic. They have negative influence on immunological mechanisms, and may cause some abnormalities. The aim of this article was to analyse smoking among pregnant, patients of Prenatal Outpatient Clinic where risk of abnormalities and defects during pregnancy is very high. 1088 pregnant, admitted to the prenatal clinic during 2005-2006, was analysed. 288 women were smokers, which made 26% of all patients. There was 23.2% high educated and 43.3% with basic education. Despite of the fact that smoking has negative influence on embryos, pregnant still are smokers. That's why antinicotine therapy should be used during pregnancy. PMID- 18409320 TI - [The study of healthy attitudes and knowledge related to smoking habit among young women living north-easterly region of Poland]. AB - The aim of presented study was to estimate the phenomenon of spreading smoking cigarettes and knowledge about it among young women living north-easterly region of Poland, which are not connected with medical or cosmetic sector. All informations was collected from the pool questionnaire, which was filled in by each person independently. The questionnaire study was curried out on 327 young women. They were quaranteed anonymity. The research touched such questions as knowledge related to number of harmful substances in cigarettes, professional activity, care for appearance and keep fit, the environment of the interviewed women. PMID- 18409321 TI - [Behaviour concerning smoking among the patients making use of advice in women health centres]. AB - The level of knowledge in the society about the harmful influence of smoking is increasing systematically. But there are still many people ignoring the warnings and prohibitions concerning smoking. The results of the research show that it is highly worrying that there are people for whom smoking is incredibly dangerous, e.g. children, youth, women, especially pregnant women. The aim of the work was to establish the percentage of smoking women among the patients of the women health centre, with the special focus on pregnant women. There were 120 women encompassed in this study in the health centre in Opoczno and 120 women using a similar health centre in Lodz between the 1st and the 15th March 2007, using a auditoria survey questionnaire. The collected data was worked out statistically. In the group of 240 tested people, 87 admitted to smoking, which is 36.3% of the respondents. Among the 185 women who were not pregnant, but were smoking, there were 75 (40.5%) and in the group of 55 pregnant women, there were 12 who smoked (f=0.22). Over 22% of the smoking women smoked over 10 cigarettes a day. From among 87 of the surveyed, 35.6% claimed they smoked everywhere they wanted. Majority of the respondents that is 52.9% lived with at least one other smoking person. Over 70% of them would like to quit smoking. Almost 48% stated their doctor has never talked with them about the influence of smoking on their health and almost 42% stated that no nurse or midwife has ever talked to them about this subject. Frequency of smoking among the tested people who were using the women health centre was high. Especially worrying was the percentage of the smoking pregnant women--every fifth of them smoked. PMID- 18409322 TI - [Cigarette smoking among nursing personnel employed in the oncology departments, in the context of the educational tasks performed]. AB - Cigarette smoking has become a popular occurrence in the 21st Cent. It was recognised by the WHO as a disease and classified as the one associated with the addiction. The list of illnesses related to smoking is still expanding while the adult smokers keep living shorter. These observations should then be reflected in creating "a fashion for non-smoking" among the health care professionals. The analysis of the extent of cigarette smoking among the nursing personnel, employed in the oncologic departments was the chief objective of the present study. The diagnostic survey was the employed method of research. It was aided by a questionnaire. The studied group comprised 100 male and female nurses, randomly selected, and employed in the oncologic departments. The survey was performed between April and June, 2007. The return rate was 71%. The results will be confronted with the earlier assessment done by the male and female nurses. The study revealed that the majority of the nursing personnel, employed in the oncologic departments do not smoke. The main reasons behind the cessation of smoking by the respondents, the former smokers, were: health considerations (45.5%), and the social ones (36.4%). It was also established that the personnel are aware of the influence of the demonstrated behavioural patterns, both conducive and detrimental to health, on the fact of being credible during a health education session, performed by them. Cigarette smoking by nursing personnel, employed in the oncologic departments is not a widespread occurrence. The personnel should, as often as possible, direct their educational actions on the patient and his/her family, and emphasise the influence of the proper life style on the preservation of health, its improvement, and the prevention of diseases or complications. Alerting the nursing personnel to the association between the behaviour patterns of the therapeutic team, and the behaviour patterns and attitudes conducive to health of their patients, is important. PMID- 18409323 TI - [Life competencies and smoking among teachers]. AB - Smoking cigarettes is one of the most common factor disturbing health and a part of negative life style. The aim of the study was to estimate the frequencies of smoking cigarettes and their conditions among teachers of all types of schools- primary as well as secondary. 94 persons participated in the study. 20.2% of them were smoking and almost 80% were not smoking. 27% persons smoked in the past. Psychologicale test standarised and normalised and author's questionnaire of own concept used in the study. RESULTS: 74% smoking women did it after breakfast, 53% at working place. 52.6% women stated that stress is a main reason of smoking and 36.8% used cigarettes after an eating. Almost 63% smoke during brakes at work and in social circumstances. Nie przerywajac pracy pali 10.5% of respondents smoked during work activities and 42% during resting time. According to opinion of examined group, the joy, relax are the effects of smoking. There were no significant differences and correlations between fear as a state and character and smoking. However the frequency of smoking in the group was lower in comparison to medium values in population of central part of Europe in a group of women. PMID- 18409324 TI - [Can smoking habit be a real risk factor of coronary heart disease in firefighters?]. AB - Active firefighting is strongly associated with exposure to health and live threads (smoke, toxic substances, high temperature) and stress (human suffering, injury, death). Tobacco smoking as a form of stress reduction is an important social and health problem in this group. Smoking as one of the well recognized cardiovascular risk factors may be associated with a risk of death from coronary heart disease in firefighters. The group of 174 active firefighters from wielkopolska region was examined based on Fagerstrom's and Scheiner's questionnaire. 46% of examined firefighters were active smokers. Mean age of smoking initiation was 16 years, motivation was indicated as an influence of school fellows, curiosity and social situations. Smoking was continued mainly due to professional stress, influence of friends and social situations. 99% considered smoking as harmful and 84% expressed high motivation to quit smoking. It seems to be of special importance to undertake prophylactic measures for oligosymptomatic cardiologic problems and creation of special programs allowing to maintenance of physical fitness among firefighters. Another area of concern should be psychological education on stress-lowering techniques and on general knowledge on a harm of smoking. PMID- 18409325 TI - [Selected socio-medical factors influencing health awareness of the old persons in the area of tobacco smoking]. AB - The aim of the research is a subjective health assessment by the old and their level of health awareness. The research was carried out by means of the SF-36 survey (abridged version) done to examine 71 old people (47 women, 24 men), former smokers or sporadic smokers. The SF-36 scale was employed to evaluate health condition in the area of physical efficiency, psychosocial functions and patients' general opinions about their health. Most of the examined were people with low health self-assessment, health (locomotive disability) and economic problems. The insufficient health awareness level of the examined elderly people indicated a necessity of implementation of education programmes and easy access to health promotion institutions. PMID- 18409326 TI - [Influence of tobacco products' advertisements on behaviour of the 'Quit and Win' competition]. AB - Smoking cigarettes, pipes or cigars is in fact inhaling harmful tobacco smoke that is created as a result of burning. Harmful substances that are part of this smoke get inside all the organs, upsetting their activities and the proper running of the life processes. In many countries, spreading the habit of smoking has caused the unwanted changes in the health state of the people. This fact does not prevent the tobacco concerns from tricky advertisement of their products. In the work there have been presented the opinions of the participants of the 'Quit and Win' competition concerning the influence of promotion and advertising of tobacco products on their smoking behaviour. The subject of the analysis are the answers received through the postal survey in June 2001 from the 900 participants of the 'Quit and Win' competition (52.9% of all the participants) organized in the region of Lodz and Kalisz at the end of the 2nd International Antinicotine "Quit and Win" Campaign.. The result have shown that in the group of 900 respondents, 160 people (17.8%) claimed that promoting tobacco has become an obstacle in sustaining tobacco abstinence in their case, and 192 people (21.3%) did not have any opinion on that subject. Though majority of the respondents (58.1%) in the group of 900 people claims that promoting cigarettes in their case had no influence on their decisions concerning smoking, many of them are people who are of contrary opinion or are unable to make any evaluation. In the case of tobacco producers, making this effort to convince us about cigarettes being not harmful proved ineffective. Giving into the influence of the insidious cigarette advertising by the adults make lead the conclusion that frequency with which adolescent and very young people take up smoking may be a result of such promotion. Eliminating tobacco advertisements as a relevant factor leading to smoking, will enable to increase the ratio of non-smokers in the society. PMID- 18409327 TI - [Tobacco smoking in workplaces]. AB - Smoking in a workplace can be an important source of the exposure to environmental tobacco smoke (ETS). Harmful effect of ETS on the human health is well proved phenomenon. Exposure to ETS increases the risk of the cardiovascular diseases, the respiratory symptoms and illnesses, and the lung cancer. The installation of the ventilation and filtration systems or separation of smokers and non-smokers decrease the exposure to ETS but do not solve the problem. One reasonable and economically justified solution is complete restriction of tobacco smoke in closed area of the workplaces. PMID- 18409328 TI - [Chromosome alterations in tobacco smoke-associated tumors]. AB - A role of tobacco products in cancer incidence is commonly known and accepted. It is estimated that roughly 1/3 of all the cancers is resulted from previous exposure to tobacco. An impact of tobacco smoke carcinogens in formation of DNA lesions and mutations is well established. Contrary to that, less is known about rearrangements of chromosomes. Nevertheless, there are many indications associating rearrangements of chromosome arms 3p, 3q, 8q, 9p, 17p i 18q with a clastogenic activity of tobacco smoke. An evolution of cytogenetics from conventional techniques to molecular cytogenetics provides an opportunity to find some links between chromosome aberrations and activation of oncogenes as well as deactivation of tumor suppressor genes. PMID- 18409329 TI - [Tobacco intake and the risk of urogenital disorders]. AB - The association between tobacco intake, in different forms (smoking and smokeless tobacco), and frequency of urogenital disorders was analyzed. Based on wide bibliography the authors have concentrated especially on tobacco related urogenital neoplasm. Some types of neoplasm, like bladder carcinoma, were significantly tobacco dependent, others were not so clearly documented. PMID- 18409330 TI - [Methods of exact nonparametric inference for non-typical distributions of experimental data]. AB - In medical research we frequently find data sets with specific structure such as small data sets, unbalanced, sparse or heavily tied. The peculiar properties of those sets influence the p-value which quantity is used in decision making process. Four examples of experimental data, for which estimation of asymptotic p value leads to erroneous decision, are presented. PMID- 18409331 TI - [Do calcification of placenta reveal only maternal cigarette smoking?]. AB - Maternal smoking during pregnancy is associated with not only obstetrical and pediatrician complications with fetus, newborn and child but also with placenta. The aim of the work was the estimation of placental calcification between maternal cigarette smoking and non-smoking pregnant women. All women have taken vitamin supplements during time of experiment. The study group consisted of 200 pregnant women (100 smokers and 100 non-smokers). All patients were admitted and delivered singleton births in the Gynecological and Obstetric Clinical Hospital in Poznan, between 2004-2007. A questionnaire was used as a study tool. s. The study groups were similar under age, education, obstetrical interview and cigarette smoking (5-10 cigarettes/day). Ultrasound investigations showed placental calcification in 36% vs 45% pregnancy at 30 +/- 2 weeks of gestation and in 52% vs 57% at 40 +/- 2 weeks of pregnancy as well as in 55% vs 58% placentas in macroscopic examination, between smoking and non-smoking women. Cigarette smoking should not be considered as exclusive factor leading to formation deposits of calcium in placenta. The consumption of vitamin supplements probably also involve calcification of placenta tissue. PMID- 18409332 TI - [Smoking cigarettes and pain--implications for the postoperative period]. AB - Smoking cigarettes poses a number of relevant medical and social problems. Impact of smoking on pain threshold and tolerance may be of significance for surgical patients, who are prompted to abstain from cigarettes before operation. Association between smoking and pain perception is complex. Experimental data bring evidence for analgesic action of nicotine and tobacco smoke acting via nicotinic acetylochline receptors (nAChR). However, clinical studies are unequivocal. Smoking is connected with some pain syndromes. Smokers take much more analgesics than non-smokers and probability of developing opioid dependence is increased in this group of patients. Smokers also present with altered mechanism of stress-induced analgesia and both gender and pain modalities influence their pain perception. Some studies demonstrate increased requirements for postoperative opioid analgesia in smoking patients. Strategies for postoperative pain treatment in smokers should involve regional techniques and clonidine. PMID- 18409333 TI - [Cigarette smoking and immunity disorder]. AB - Cigarette smoking addiction is one of the cardinal factors the diseases of respiratory system, cardio-vascular system and neoplasms. A part of them is the effect of immunosupression. It includes both morphologic changes (atrophia of lymphoid tissue) and cell functional disorder of the immune system. Cell-mediated response Th1-type, which is responsible for pathogens invasion, is depleted, whereas Th2-type is increased and has its effect on higher frequency of allergic diseases. Susceptibility to neoplasmatic changes is also augmented. PMID- 18409334 TI - [Smoking-related diseases--the role of physician (pathologist) in diagnosis and preventive activities]. AB - Tobacco smoke is rich in biologically active substances. Among them several may have an harmful impact on health consequences. All tobacco-related diseases are in fact tobacco-nonspecific and might also develop in tobacco-free circumstances. In this paper there are presented some current data indicated a significant role of tobacco smoke in development of selected disease. PMID- 18409335 TI - [Risk factors for smoking relapse after delivery--review of the study results and analysis of the study conducted in Polish population]. AB - Most of women who quit smoking during pregnancy return to smoking within 12 months after delivery. Since smoking cessation is more likely during pregnancy than at other times, interventions to maintain quitting postpartum may give the best opportunity for a long-time abstinence. It is still not clear what kind of advice or counseling should be given to help prevent the post-delivery relapse. To find an answer to that question, we need to know which factors predispose the women for and which protect from restarting smoking after delivery. The profile of women who relapse to smoking can be described as living with smokers. Breastfeeding and positive experience related to the quitting of smoking protect the women from returning to smoking after delivery. Those factors should be taken into account while creating antismoking intervention. PMID- 18409336 TI - [Intervention programmes aimed at the reduction of tobacco smoking among young people: methodological issues]. AB - Tobacco smoking concerns to over one in four Polish teenagers. Many adolescent tobacco programmes focus on preventing teenagers from starting to smoke, but some programmes have been aimed at helping those teenagers already smoking to quit. Complex programmes, including those tailored to the young person's preparation for quitting, and behavioral therapy programmes show some promise. However, the number and quantity of interventions do not yet provide enough evidence to judge effectiveness. Pharmacological treatments such as nicotine replacement therapy and bupropion have not yet been sufficiently tested in adolescents. Interventions used different definitions of smoking adolescent, quitting and smoking verification therefore their results have not been comparable. Some approaches may be worthy of consideration but there is still a need to provide stronger and more persuasive evidence before large scale investment in programmes. In the polish references has not been found a description and evaluation of the complex and effective programmes including psycho-social intervention for smoking teenagers. PMID- 18409337 TI - [Varenicline--a new chance for smokers?]. AB - Cigarette smokers believe that quitting smoking is the hardest thing in their life. Modern pharmacology offers a wide range of various drugs and therapies that might support smoking cessation. One of the most effective drug is varenicline. Varenicline has been introduced this year to Polish pharmaceutical market by Pfizer company under the trade name Champix. The aim of the study was to critically review all new information about Champix pharmacology and pharmacokinetics but also evaluate safety and economical aspects of using this new drug. From the presented data it might be concluded that nowadays varenicline is the most effective drug used in smoking cessation (about 40%). The unique properties of varenicline, which is a partial agonist and antagonist to alpha4beta2 nicotinic receptor, are the reasons of dual mechanism of the drug action. The agonist effect is caused by binding to nicotinic receptors and stimulating receptor-mediated activity. The antagonist effect occurs when varenicline blocks the ability of nicotine to activate nicotinic receptors. The most frequently adverse effects of varenicline are: nausea, headache, insomnia, and abnormal dreams. Although the price of complete therapy with Champix seems to be quite high, but potential benefits (in case of therapy finished successfully) are much higher than these costs. Pharmacological properties of varenicline described in the article, along with its high relative effectiveness and safeness, make the drug very attractive and promising for the smokers who want to quit. PMID- 18409338 TI - [Chemoprevention of tobacco-related lung cancer by cruciferous vegetable]. AB - Lung cancer is the most common malignant disease in the world and the major cause of death from cancers. Around 80-90% of all human lung cancers are related to cigarette smoke. Tobacco smoke contains at least 60 carcinogens capable of causing tumors, 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and polycyclic aromatic hydrocarbons (PAH) are among the most prominent pulmonary carcinogens. Prevention is the most effective way to reduce lung cancer mortality. Chemoprevention is a cancer preventive strategy to inhibit, delay or reverse carcinogenesis using naturally occurring or synthetic chemical agents. A number of epidemiological studies have shown an inverse relationship between cruciferous vegetable consumption and cancer, especially those of lung and stomach. Crucifers, such as broccoli, cauliflower, brussel sprouts and cabbage, contain a family of secondary plant metabolites known as glucosinolates, which are unique to these vegetables. Upon hydrolysis, glucosinolates yield a number of breakdown products, mostly isothiocyanates, with supposed chemopreventive properties, as shown in animal experiments. It appears that significant portion of the chemopreventive effects of isothiocyanates may be associated with the inhibition of the metabolic activation of carcinogens by cytochrome P450s (Phase I), coupled with strong induction of phase II of detoxifying enzymes. PMID- 18409339 TI - [Turkish (Aztec) tobbaco (Nicotiana rustica L.) unnoticeable danger?]. AB - Tobacco harmful influence on men's health is usually linked with smoking cigarettes. Alternative ways of intake tobacco are also dangerous. Nicotine and tobacco-specific N-nitrosamines with carcinogenic activity are not only present in tobacco smoke but also of nicotine-rich product for chewing. There are some internet sources about Turkish (Aztec) tobacco (Nicotiana rustica L.) as easy available material for home-made tobacco mixtures. A some reports about dangerous effects connected with chewing tobacco and similar to smoking-related disorders are present in current literature. PMID- 18409340 TI - [Genetically modified tobacco--chance or threat for smokers?]. AB - Tobacco (Nicotiana sp.) is of the most often used model plants for research in the field of physiology, biochemistry, molecular biology and genetic engineering. Tobacco was also the first plant subject to genetic transformation--in 1983 the plant was introduced with the kanamycin resistance gene. The internet database PubMed includes roughly 3,500 records concerning transgenic tobacco, out of which nearly 200 works were published this year. Advancement of state-of-the-art techniques of genetic engineering offers new, as yet unprecedented opportunities to take advantage of tobacco. Apparently tobacco is not only a source of carcinogenic tobacco smoke, but also a plant which due to genetic modifications may be used in a positive way in medicine and biological sciences. The aim of the article is to summarize the role of genetically modified Nicotiana genus plants in modern medicine and environmental protection. The publication describes the usage of transgenic plants in the tobacco industry and evaluates possible effects of such applications. Moreover, potential possibilities of deployment of genetically modified tobacco for production of anti-virus and antibacterial vaccinations, interferon, antibodies, etc. have been characterized. PMID- 18409341 TI - [A persistence of tobacco smoking addiction in economically severely underdeveloped regions of Poland--the case of the Green Forest and White Forest]. AB - In the 19-th and early 20-th centuries the regions of the Green Forest and White Forest (Central Poland) comprised a reservoir of rural people living on the verge of biological existence. Despite material deficiencies, tobacco smoking addiction was common--nearly 100% of the male population was smokers. An analysis of ethnographic findings suggests that the permanence of addiction could have been caused by 1. the early initiation of smoking and acceptance of smoking by under ages, 2. the spreading of the practice of replacing tobacco, when in deficit, by other materials such as dried potato leaves, clover leaves or even feces of horses. One can assume that stimuli linked with non-tobacco-exposed smoking may also have an important influence on maintaining addiction. PMID- 18409342 TI - [Diagnostic strategy and prognostic significance of syncope in patients with organic heart disease]. AB - Chronic heart failure (CHF) is a very serious problem in the health care systems of all developed countries. Immense progress that has been made in medical sciences for the last twenty years, has not only caused remarkable lifespan prolongation, but has also resulted in increasing number of patients suffering from CHF. One of the most common chronic heart failure symptoms is syncope. Pathophysiologic background of this phenomenon is complicated and frequently connected with heart failure cause. It has been proved in a few studies that syncope in CHF patients is a unfavorable prognostic factor and can be a predictor of sudden cardiac death. Patients demonstrating syncope and suspected of organic heart disease should undergo echocardiographic examination, prolonged electrocardiographic monitoring and if necessary - electro-physiological study. Patients reporting chest pain suggestive of ischeamia before or after loss of consciousness should udergo an electrocardiographic stress test as a first evaluation step. The most probable mechanism of syncope in patients with structural heart disease is arrhythmia. It has been proven that patients with heart disease-related syncope have higher mortality rates than population without structural heart abnormalities. Higher incidence of sudden cardiac death in CHF patients experiencing syncope should not be forgotten by the physician. All causes of syncope should be carefully diagnosed, especially in this population. It may help to administer appropriate treatment and reduce sudden cardiac death risk. PMID- 18409343 TI - [Effectiveness evaluation of knee joint 90Y radiosynovectomy]. AB - INTRODUCTION: Chronic knee synovitis with effusion, because of the special role of knee joint is an important therapeutic problem. This leads to searching for new treatment modalities. One of them is radiosynovectomy based on anti proliferative and anti-inflammatory activity of ionizing radiation. It is made using 90Y mainly (high energy of beta [electrons] radiation [2.2 MeV], large average penetration in soft tissues [3.6 mm] and long physical half-life [2.7 days]). MATERIAL AND METHOD: Analyzed material is comprised of 30 patients (33 treatments) suffering from proliferative synovitis of knee joint treated by radiosynovectomy using intra-articular injection of 6 mCi 90Y. In 20 cases the reason of disease was non-specific reactive arthritis, in 5 rheumatoid arthritis, in 3 villonodular synovitis, in 3 psoriasis and in 2 ulcerative colitis. Symptoms duration varied from 3 to 144 months (mean 43). A knee circumference at the treatment day varied from 33.5 cm to 49 cm (mean 41). The operation was based on knee biopsy, evacuation of exudate and delivery of 6 mCi of colloid 90Y. Follow up ranged up to 14 months (mean 4.4). Patients were examined 2 weeks, 1, 3, 6 and 12 months after treatment. During examination a knee circumference was measured, a knee mobility, temperature and patella floating symptom were examined. Pain in treated region was assessed and amount of exudate was measured. RESULTS: A knee circumference and exudate amount enlarged during following controls, and compared between the treatment day and the last control did not differ significantly (41 vs. 41.6 cm and 43 vs. 42.5 ml respectively) but number of biopsies decreased. A percentage of patients with impaired knee mobility also did not change (59% vs. 58%). Probably, it was caused by resignation from control examination when symptoms disappeared. A percentage of patients without pain relief decreased from 43.5% 2 weeks after treatment to 20% one year later and a patient percentage with complete pain relief increased from 8.5% to 60%. A patient percentage with increased knee temperature and with floating patella decreased significantly (54 vs. 25% and 83 vs. 48% respectively). CONCLUSIONS: Obtained results do not allow to form univocal conclusions regarding effectiveness of 90Y radiosynovectomy. The decreasing biopsies number, decreasing percentage of patient with increased knee temperature and with floating patella; increased percentage of patients with analgetic effect and with total pain relief show a necessity of renewed evaluation of this treatment modality on the base of bigger patients number and longer and more precise observation. PMID- 18409344 TI - [Stereotactic extracranial radiosurgery of liver tumors using respiratory gating- the method presentation]. AB - Radiotherapy has not been widely accepted in the treatment of primary and secondary liver tumors due to its limited efficacy. This is caused by insufficient radiation doses and suboptimal techniques, which were used in the past. However, a present technological progress in radiotherapy has led to the application of modern and sophisticated radiation techniques into the treatment of liver tumors. The excellent example of such progress is stereotactic body radiation therapy (SBRT) (or radiosurgery), with respiratory gating. Such treatment method enables delivery of high radiation doses precisely to the tumor with sparing of surrounding critical tissues. It is possible by application of multiple beams and a narrow margin around the tumor considering its respiratory movement. This method of treatment may be applied under the condition of very precise radiological imaging and accurate patients' immobilization. The study presents a method of stereotactic body radiation therapy with respiratory gating introduced in the MSC Cancer Center and Institute of Oncology in Gliwice. The treatment criteria are as follows: patients in good general condition with inoperable primary and secondary tumors, 1-3 lesions with a maximal diameter of 6 cm. The fractionation schedule consists of three doses of 12 Gy delivered in weekly intervals to the total dose of 36 Gy normalized to the minimum in the PTV. Pre-treatment preparation procedures included: patient immobilization in a vacuum pillow, monitoring of respiratory cycle, the choice of respiratory phase suitable for irradiation and computed tomography (CT) for treatment planning. After preparation of treatment plan, the X-ray simulation is performed with verification of the position of isocenter. A respiratory tracking is performed during simulation. At the last stage, patient is immobilized at the couch of linear accelerator, the verification of the isocenter position using MV or kV images is performed and respiratory tracking is once again conducted. Irradiation delivered intermittently in the selected respiratory phase using automatical connection between respiratory tracking system and accelerator. The SBRT with respiratory gating appears to be a promising method of local treatment in patients with inoperable liver tumors. PMID- 18409345 TI - [Own experiences in treatment of the external and middle ear tumors from data collected between 1992-2001]. AB - We discuss a retrospective study of patients with malignant tumors of the external and middle ear operated at the Otolaryngology Department of the Jagiellonian University in Cracow between 1992-2001. We performed 56 operations of malignant lesions in that period of time. We evaluate primary localization of the tumors, local progression, histopathological recognition, performed treatment and distant results of therapy. In our population the most common tumor was basal cell carcinoma located on the auricle in T1 stage. The most common lesion located in the middle ear or middle and external ear was the squamous cell carcinoma in T2 stage. Only surgical treatment was performed in about 60% of patients. 40% required combined therapy with surgery and postoperative radio and chemiotherapy. 80% of patients survived 5 years after performed treatment. PMID- 18409346 TI - [Mean number of surgical interventions and number of engaged physicians' in general surgical departments in different health care systems]. AB - AIM: We examined some data from surgical departments during health care reform (1998-2004) in two different regions of Poland. MATERIAL AND METHODS: We analyzed number of surgical interventions and number of engaged physicians' in 60 surgical departments in two different regions of Poland from 1998 to 2004 during health care reform. RESULTS: We showed significant systematical increase of number of surgical intervention in major surgical departments (more than 30 beds). In minor surgical departments (less than 30 beds) increase number of surgical interventions were observed only to the year 2001. We didn't showed changes in number of engaged physicians' in the analyzed period. We observed that ratio: number of surgical intervention/number of engaged physicians' increase only to the year 2002. CONCLUSION: This preliminary data showed beneficial trends in health care during the reform. PMID- 18409347 TI - The evaluation of the effectiveness of multiple dose intradermal hepatitis B re vaccination in hemodialyzed patients not responding to standard method of immunization. AB - Hepatitis B is a serious epidemiological problem in uremic patients treated with renal replacement therapy. A high proportion of hemodialyzed patients do not respond to the standard method of intramuscular (i.m.) hepatitis B vaccination. Low-dose intradermal (i.d.) inoculations and supplementary i.m. injections have been reported to improve the responsiveness in formerly non responding uremic patients. We applied a inoculation schedule of 10 microg Engerix B i.d. in 49 pts and i.m. (control group) in 13 pts once a week during 12 consecutive weeks in order to compare the effectiveness of the various ways of immunization in maintenance dialyzed patients not responding to standard vaccination. Serum anti HBs antibody level, as well as biochemical and immunological parameters were examined. Already one month after initiation of the cycle, 57.1% of patients in the i.d. group responded by achieving the minimum protective anti-HBs antibody level (>10 IU/I.); while 14.3% reached full adequate anti-HBs antibody level (>100 IU/I.). After the full therapy period, anti-HBs antibody level >100 IU/I. was achieved in 42.9% of the patients, while a total of 81.7% of patients reached the anti-HBs antibody level >10 IU/I. In 18.4% of patients no response was observed. Surprisingly similar results were achieved in the i.m. group. Twelve months after termination of the inoculation cycle we noted decrease of anti-HBs antibody level; the values >100 IU/ I. was observed only in 18.4% of the study group, while 87.8% reached a titre >10 IU/I. We found a relationship between the effectiveness of immunization and RBC count, total serum protein and albumin levels and GGTP activity. Mitogen stimulation indexes in both groups were 4-5 times lower in comparison to reference values in the general population. In the study group that did not respond to vaccination, mitogen stimulation indexes were 2 times lower as compared to the group characterized as having a good response. In conclusion, the route of injection seems to be less important than the frequency and number of doses of the vaccine. Anemia and malnutrition may be responsible for the worse response to vaccination against hepatitis B virus. PMID- 18409348 TI - The influence of sodium profiling on blood volume and intradialytic hypotension in patients on maintenance hemodialysis. AB - An accurate evaluation of water content in the body of hemodialyzed patients seems to be an important problem in chronic dialysotherapy. Acute intradialytic hypotension observed in 20-33% of hemodialysis patients is a very common complication of this kind of renal replacement therapy. The study was performed in 40 uremic patients, treated with hemodialysis at the Nephrology Clinic of the University Hospital in Cracow. In every patient, 3 model dialysis sessions were carried out. Total fluid removal was the same during every hemodialysis. The first model hemodialysis (HD1) was performed with constant dialysate sodium concentration (140 mmol/L), the second (HD2) with linear and the third (HD3) with expotential decrease of dialysate sodium concentration (from 144 to 136 mmol/L). Every hemodialysis was also monitored continuously with Crit-Line 2 R system (In Line Diagnostics, Riverdale, UT). Before and after the first model hemodialysis (HD1), ultrasound examination of abdominal cavity was performed. The measurement of inferior vena cava diameter (mm), circumference (mm), area (mm2), at hepatic veins orifice-level, on expiration was performed. The fluid removal during the first model hemodialysis resulted in significant reduction of the vena cava inferior diameter, circumference and area, measured with ultrasound on expiration. The statistically significant lower fall of blood volume after the first and second hour of the second model hemodialysis session (HD2) was observed when compared to the first hemodialysis (HD1) - p<0.05 was observed. The statistically significant lower frequency of hypotension during the second hemo dialysis session (HD2) as compared to HD1 (chi2=5.25 p<0.05). Differences among HD1 and HD3 and HD2 and HD3 did not reach statistical significance. The monitoring of hemodialysis with the Crit-Line instrument permits for optimalization of dry weight of dialyzed patients and allows reaching higher ultrafiltration rates during dialysis without hypotensive episodes. The changes in the blood volume, approximately 5% per one hour of dialysis session are an optimal value for these patients. PMID- 18409349 TI - [Transferrin microheterogeneity in patients treated by maintenance haemodialysis]. AB - Transferrin (Tf) plays a crucial role in iron transport in the human body. In physiologically status Tf occurs in four variants. The concentration of Tf and its variants is changing in several conditions, especially during acute phase response, which may take place in end-stage renal disease (ESRD). The aim of this study was the assessment of the associations between changes of glycosylation of transferrin and selected blood cell count and iron metabolism parameters in ESRD patients treated by maintenance hemodialysis (mHD) with long lasting anaemia despite treatment by rHuEPO. Studies were carried out on a group of 55 ESRD patients treated by HD (ESRD group). The reference values were obtained from a group of 20 healthy volunteers (HV group). The Tf concentration was determined by the rocket immunoelectrophoresis according to Laurell. To estimate the microheterogenity of Tf the crossed affinoimmunoelectrophoresis according to Bog Hansen was used. The concentration of selected hematological variables, and selected liver metabolism variables was assessed by routine laboratory tests, while the concentration of cytokines was measured by ELISA method, and selected acute phase proteins, i.e. C-reactive protein, by immunonephelometry method. Tf in studied sera was divided into four variants Tf1, Tf2, Tf3, Tf4. The percentage contents of variants in HV was Tf1=4%, Tf2=9%, Tf3=82%, Tf4=4%. In the sera of ESRD group the percentage contents of the Tf1, Tf2, Tf4 variants increased to 9%, 16%, 10% respectively, while the Tf3 variant decreased to 65%. Serum concentration of Tf was lower (1441 +/- 526 mg/L) in ESRD patients (p<0.0001) vs. 3539 +/- 955 mg/L in HV group. During this study we have revealed that the quantity and the quality of Tf is changing in ESRD patients. The observed correlations between concentration of Tf3 and selected hematological variables and also between Tf3 and rHuEPO, suggests that the decrease concentration of the Tf3 may lead to an impaired iron transport to the cells and may be one of the factors affecting anaemia in patients treated by HD and rHuEPO. PMID- 18409350 TI - [Peritoneum as a specific membrane]. AB - This paper is a review of the literature describing the complexity of the peritoneal membrane structure and functions, mechanisms of the peritoneum failure as a dialyzing membrane and strategies for its protection during prolonged peritoneal dialysis. PMID- 18409351 TI - [The role of nitric oxide (NO) in acute pancreatitis]. AB - The hypothesis of the role of nitric oxide (NO) in acute pancreatitis (AP) is variable. According to some authors, increased level of nitric oxide has a positive influence on the course of AP. On the other hands, others show negative role of NO in this disease. NO plays many different roles in the organism. It regulates blood pressure, relaxes smooth muscle, plays a role in neurotransmition (for instance in specific and nonspecific immunological responses). NO regulates microcirculation within the pancreas and has an impact on the clinical courses of the mild as well as severe forms of the disease. It is also produced in the lungs alveoli and it is partly responsible for pulmonary complications of AP. PMID- 18409352 TI - [The conjugated linoleic acids in prevention and treatment of obesity]. AB - The results of up-to-date performed experimental studies indicated that conjugated linoleic acids (CLA) contained in diet lead to significant both adipose tissue and body mass decrease. It suggests that the CLA-rich diet or CLA contained medicines can be useful in prevention and treatment of obesity in humans. The CLA-contained medicines are more and more popular, therefore the principle aim of this review is a description of CLA chemical structure, presence in natural products and moreover, their potential influence on adipose tissue mass in humans. Unfortunately, the results of recent trials are disappointing. Most of them indicated that (contrary to experimental studies) CLA were not profitable in adipose tissue mass decrease in humans. Moreover, in a few cases a deterioration of serum lipid profile, glycemia enhancement and decrease of tissue insulin sensitivity were observed. Thus, there is no evidence confirming the positive influence of CLA on adipose tissue mass in humans. However, there are also a few publications, in which the CLA-associated decrease of adipose tissue mass in humans was noted. The results of these studies do not let us totally decline the opinion that CLA are profitable also in humans. This contradictory data definitely needs further studies. PMID- 18409353 TI - [Mechanisms of nickel allergy]. AB - Nickel allergy constitutes a serious health problem of modern societies. Hypersensitivity to this metal is found in 13% adults and 8% children. Risk factors for nickel allergy are: female gender and early exposure to nickel, e.g. piercing. Various mechanisms of inducing nickel allergy are possible, which is also reflected in the different clinical pictures. Nickel can induce allergic reaction in 3 different ways: 1) it binds to carrier protein in the extracellular space and subsequently is processed and presented by antigen presenting cell (APC) in the context of MHC class II molecule, which activates CD4+ lymphocytes, 2) Ni penetrates into the cell where it binds to intracellular proteins, and subsequently it is presented in the context of MHC class I molecule, which activates CD8+ lymphocytes, 3) Ni can "bridge" MHC molecule together with the TCR receptor on lymphocyte without actually filling the antigen-binding site, which is in analogy to superantigens. Both Th2/Tc2 (IL-4, IL-5, IL-13) and Th1/ Tc1 (IFNgamma) take their part in the development of contact allergy to nickel. The trafficking of the effector cells to target organs (where the inflammatory reaction actually takes place) is controlled by homing antigens and chemokine receptors that are expressed on their surface. The accumulation of effector cells in a target organ can determine the symptoms of nickel allergy (the skin, mucosa etc.). The acquisition of nickel tolerance is possibly dependent on the IL-10 secretion by specific lymphocytes. PMID- 18409354 TI - [Type IV of hypersensitivity and its subtypes]. AB - Type IV of hypersensitivity reaction is usually manifested in the skin in different clinical pattern. According to traditional Gell and Coombs classification, the mechanism of IV type of allergic reaction has been associated with contact allergy with the activity of lymphocytes Th1 secreting interferon gamma. Now, this vision seems to be too simplified. In the last years there were publications, which can throw a new light on these complicated mechanisms leading to the development of the type IV of allergy, especially to drugs, nickel and other haptens and also can explain the differentiation of clinical pattern in respective patients. The skin symptoms in type IV of hypersensitivity are triggered by activation of specific T-cell CD4+ and CD8+. Immunohistochemical and functional analysis of reactive T-cell has shown that the delayed hypersensitivity reaction depends on the secreted cytokines. For example maculo papular exanthema may be either triggered by Th1 or Th2 in nature and cytokines interferon gamma, tumor necrosis factor alfa or interleukin-4, 5 and 13. Bullous reactions (i.e. Stevens-Johnsons Syndrome or toxic epidermal necrolysis) are characterized by widespread keratinocyte apoptosis, a consequence of high CD8+ T cell involvement and the molecular cytotoxicity of Fas, perforin and granzyme B. Pustular exanthema reactions are stimulated via the T-cell release of 11-8 and granulocyte-monocyte colony-stimulatig factor (GM-CSF). For the better understanding of these inflammatory cascades deleted type IV of hypersensitivity reactions have been re-classified into four main subtypes: 1. IVa with Th1 and monocyte directed and cytokines: IFNgamma, IL-1, IL-2, 2. IVb with Th2 and eosinophils directed and cytokines: L-5, IL-4, IL-13, 3. IVc with T CD8+ directed and cytokines: perforin, granzyme B, Fas Ligand, 4. IVd with T CD4+, CD8+ and neutrophil directed and cytokines: IL8, GM-CSF. Clinically delayed hypersensitivity eruptions are often an overlap of cytokine pathways, with one preferential reaction dominating the final picture. Type IVa and IVc play a role inthe mechanism of contact dermatitis, however type IV b in chronic asthma, chronic allergic rhinitis and maculo-papular exanthema with eosinophilia, type IV c in bullous reactions (i.e. Stevens-Johnsons Syndrome or toxic epidermal necrolysis), so type IV d in pustular exanthema reactions (i.g. AGEP - Acute Generalized Exanthematosus Pustule, Behcet disease). This different clinical pattern of allergic disease mainly including drug allergy to nickel and other haptens as well as chronic asthma and allergic rhinitis may be explained by above mechanisms. The study of different mechanisms of four subtypes of type IVof allergic reaction may be helpful in the differential diagnostics and in the treatment of allergic diseases. PMID- 18409355 TI - [Drug-induced lupus erythematosus and systemic lupus erythematosus--differences and similarities]. AB - Drug-induced lupus (DIL) is a lupus-like syndrome. Its clinical and laboratory features are very similar to idiopathic lupus. First description of DIL is from 1945. Since that time over 80 medications are known to be responsible for development of the disease. Nevertheless mechanisms responsible for this process are not known enough. Nowadays direct and indirect mechanisms in DIL are known. The slow acetylator status has an important position. Moreover the role of estrogens and anti-TNF-alpha therapy are taken into consideration. The T cells' participation and the methylation of DNA and their role in graft-vs.-host disease, which is the murine model for DIL, are considered among others. Researches point also to the LFA-1 and the IL-6 as factors responsible for DIL's development. Another element which could be significant in this process is the presence of antimielo-peroxydase antibodies. Also the role of disruption of T cell maturation in the thymus induced by the drugs' metabolites may have an important position. Nevertheless it is still very hard to say which of those mechanisms is the main one. In spite of everything it's not clear enough if the observed clinical manifestations are caused by drugs/their metabolites,- DIL or that they are consequences of a induction of silent process leading to SLE development. PMID- 18409356 TI - [Hypophysitis]. AB - Hypophysitis is a rare endocrine disorder with a female predilection affecting mainly young women during late pregnancy and in the postpartum period. The clinical, histopathological and morphological findings and the association of the disease with other autoimmune disorders allow most cases of hypophysitis to be included among the autoimmune diseases. This potentially life-threatening condition should be suspected especially in women of reproductive age who present with hypopituitarism or evidence of pituitary mass-induced headaches and visual symptoms. The natural history of hypophysitis is variable. At the present time, the treatment is only symptomatic but there is no absolute agreement among endocrinologists about the optimal management of this condition. In this review, aetiology, symptoms, clinical classification, diagnosis and treatment of hypophysitis are discussed with a special emphasis on the most recent literature. PMID- 18409357 TI - [Antiphospholipid antibodies with HCV infection. Innocent proteins or risk factor?]. AB - Infection with hepatitis C virus (HCV) may be associated with a wide spectrum of immunological abnormalities. HCV tends to induce nonspecific autoimmune reactions, as demonstrated by the high prevalence of various autoantibodies, including antiphospholipid antibodies (aPL). The aPL antibodies (lupus anticoagulant and anticardiolipin antibodies) are a heterogeneous family of immunoglobulins reactive with complexes of phospholipids and plasma proteins (cofactors). The most important of these protein cofactors are beta2-glycoprotein (beta2-GPI) and prothrombin. The antiphospholipid syndrome (APS) is an autoimmune disorder characterized by arterial or venous thrombosis, recurrent fetal losses in association with the presence of antiphospholipid (aPL) antibodies. Increased prevalence of aPL antibodies in several bacterial, parasitic, and viral infections have been reported. Most of the published data agree that anticardiolipin antibodies are frequently found in patients with chronic HCV infection, but they do not appear to be of clinical importance. Some studies, however, have found an increased incidence of thrombotic disorders in patients with chronic hepatitis C virus (HCV) who manifest aPL positivity. More prospective, long-term studies are required in order to address whether HCV is involved or not in the etiopathogenesis of APS. PMID- 18409358 TI - [Diagnostic and treatment processes and the requirement of patient's consent in doctors' opinions]. AB - Contemporary medicine, together with the increase in invasiveness into the patient's body, exerts mounting pressure on state institutions and medical chambers aiming at the development of suitable regulations protecting patients against excessive interference with their bodies and psyche, and also allowing for their full autonomy. A gradual shift in the doctors' behaviour from a paternalistic approach towards partnership becomes more apparent. Its purpose is to take shared decisions concerning diagnostic methods and treatment. This process is best noticed when it comes to the issue of taking medical measures after obtaining patient's conscious agreement. The term 'conscious agreement' emerged in the legal terminology fairly late, in 1957, during the lawsuit Salgo vs. Leland Stanford Junior University Board of Trustees. Much later it found its place in the legal regulations concerning everyday medical routines. The fundamental factor of the patient's autonomy and simultaneously a remedy to the risk of patients acting against their own good is the process of obtaining the sick person's conscious agreement prior to anticipated medical measures, both therapeutic and diagnostic. The patient's informed conscious agreement to diagnostic process and treatment is not only the crucial prerequisite of doctor's lawful behaviour but also a significant evaluation criterion with regard to doctor's deeds and intentions. We requested doctors to share their opinion, in the form of an anonymous feedback form, on the issue of obtaining from patients their conscious agreement to diagnostic and therapeutic measures. The survey was carried out among 231 physicians of different specialisations from the Lublin Province and Podkarpackie Province employed both in hospitals, outpatient clinics and in health centres for basic medical care. Almost all the respondents declared the knowledgw of the regulations on the protection of patients' rights; however, only half of the surveyed persons expressed the opinion that the regulations concerning patient's rights refer also to diagnostic examination. Doctors clearly differed with regard to the meaning of the notion of 'giving conscious consent by the patient' and tended to associate such a patient's declaration only with invasive procedures. The survey results allowed us to outline the level of knowledge and the degree of compliance with patient's rights related to the necessity of obtaining conscious consent to medical treatment. PMID- 18409359 TI - [Severe, life-threatening renal anemia treatment in patients who do not accept heterologous blood transfusion for religious reasons--case report]. AB - The management of severe anemia in patients who do not accept heterologous blood transfusion for religious reasons presents many different challenges both at the level of treatment strategy and ethics. Recently, new interventional treatment strategies have become available, including human and bovine hemoglobin substitutes and high-dose recombinant human erythropoietin. We present the successful management of two Jehovah's Witnesses patients with severe, life threatening anemia caused by chronic renal failure and exacerbated by sepsis. PMID- 18409360 TI - [Sarcoidosis--rare clinical form]. AB - The paper presents a case of a 46-year old male patient in whom numerous asymptomatic round shadows in both lungs were diagnosed. The pulmonary diagnostic tests confirmed the diagnosis of neoplastic dissemination of unknown origin. The broader internal diagnostics did not confirm a neoplastic cause of the changes. The histopathological examination of the supraclavicular lymphonodus lead to the diagnosis of chronic, III degree sarcoidosis including the involvement of the lung parenchyma and peripheric lymph nodes. The lung radiography - atypical for sarcoidosis (owing to the lack of hilus lymphadenopathy) - was the cause of the false primary diagnosis. PMID- 18409361 TI - [Lung tuberculosis in a patient with rheumatoid arthritis--case report]. AB - In 1993 WHO announced the tuberculosis threat, as 1/3 of the world's population was infected with Mycobacterium tuberculosis. Only in 2002 there were 9 million new cases recorded and yearly mortality rate of this disease was 3 million. Comparing to the European Union countries such as France, Germany, or Great Britain, Polish statistics show a two times higher incidence rate for 100000 people. In 2004 the greatest number of new cases was recorded in the following regions: Mazowiecki, Silesian, and Lodz district. As screening was abandoned, general practitioners play an important role in detecting this disease. The authors of the paper present a case of a man aged 67 with atypical rheumatoid arthritis and tuberculosis recidivation. PMID- 18409362 TI - [Thrombolysis during cardiopulmonary resuscitation--own clinical observations- case report]. AB - Thrombolytic therapy is contraindicated in the course of cardiopulmonary resuscitation (CPR). If the primary cause of cardiac arrest is myocardial infarction or massive pulmonary embolism, fibrynolysis may be life-saving. We present a case report of a woman admitted to the Intensive Care Unit with cardiac arrest with symptoms suggesting myocardial infarction or pulmonary embolism. After unsuccessful conservative CPR a single dose of 500000 IU streptase was administered. Heart action returned 10 minutes later, nevertheless the patient needed mechanical ventilation and circulatory system stabilization therapy (catecholamines) in doses dependent on haemodynamic parameters. During hospitalization she regained consciousness. She presented no neurological defects and after 5 days was discharged to the Cardiology Department. Electrocardiography and echocardiography done after successful resuscitation was specific to infero lateral myocardial infarction. Although safety and efficacy of thrombolytic therapy at resuscitation was extensively studied, this procedure is still controversial. Till now, there is no data concerning thrombolytic treatment in such clinical situations, which are based on clinical trials, and such treatment is introduced in dramatic situations, as a last, lifesaving option. PMID- 18409363 TI - [Surgical treatment of the lower extremities varicose veins throughout the ages]. AB - The authors describe a historical review of surgical treatment of varicose veins from antiques to modern methods. Diseases of the veins have been recognized and attempts of their surgical resection have been described since antiquity. Hippocrates wrote about surgical treatment of varicose veins, Celsus and also Galien himself described varicose veins ligatures. The end of 19th century with Trendelenburg and Madelung was a start of modern period of varicose vein surgery. Parallel to surgical the less invasive methods of varicose veins treatment developed and gave a base for modern sclerotherapy. PMID- 18409364 TI - [Centrosome as "a brain" of an animal cell]. AB - Centrosomes are the major centre of microtubule nucleation and microtubule minus ends concentration in animal cells. Microtubule plus-ends are directed to a nucleus and chromosomes or to a cell cortex. The crossing of signal transduction pathways and the network of interactions between signal molecules controlling cell cycle are revealed in centrosomes. The ability of centrosomes for reduplication suggests the existence of hypothetic template elements. It is attractive to suggest the essential role of specific centrosome-associated RNAs in biogenesis of centrosomes. Untranslated RNAs playing a structural role and mRNAs that are localized in centrosomes to regulate protein synthesis in close proximity to mitotic apparatus may be present among these RNAs. Centrosomes positioning plays the important role in determining of cell polarity. Centrosomes are critical for the formation and support of cilia and flagella having motility or sensory functions. PMID- 18409365 TI - [The history of myxosporean (Myxozoa Grasse, 1970, Myxosporea Butschli, 1881) life and nuclear cycles studies]. AB - The paper presents a historic review of various hypothesis concerning the myxozoan life and nuclear cycles. The comparison of DAPI- and Feulgen-image cytometry results of DNA amount in myxozoan actinospora and myxospora nuclei, in connection with the new data on the animal life and nuclear cycle, has been performed. Possible reasons for the data discrepancy are considered. The further perspectives of myxozoan biology, cytology, karyology and taxonomy investigation in Russia are discussed. PMID- 18409366 TI - [Chromatin structure within sites of DNA replication]. AB - Conformational changes in chromatin structure are nowadays the object of intensive research due to its importance for proper regulation of intranuclear processes. The fine structure of chromatin within the DNA replication sites was studied in in situ fixed cells and cells permebilized by low ionic strength solutions in the presence of divalent cations. The latter method provides visualization of higher level chromatin structures such as globular chromomeres and chromonema fibres. Nascent DNA was detected immunochemically using anti-BrdU antibodies on the surface of ultrathin sections prepared from Epon-embedded material. It was shown that newly replicated DNA preferentially localized within the zones filled with globular and fibrillar elements with characteristic diameter of 30 nm, and not in chromonema fibres, while after replication had been completed DNA became embedded into as thick as 60-80 nm chromonema elements. The results obtained are discussed in the context of conception of hierarchical folding of chromatin fibers. PMID- 18409367 TI - [Estimating of changes in the amyloid and prion content of yeast cells]. AB - An attempt was made at estimating the overall amyloid content of yeast cells by treating crude cellular lysates with thioflavin T, the agent specifically staining amyloid fibrils. We demonstrated that overproduction of the yeast chaperone Hsp104p, as well as GuHCI treatment of the [PSI+] cells led both to elimination of the [PSI+] factor and to a stable decrease of the overall amyloid content estimated by intensity of fluorescence (IF) of the thioflavin T. At the same time, overexpression of gene SUP35, coding the protein prionizable to [PSI+], led to generation of [PSI+] clones with higher IF of thioflavin T. Cytoduction in the crosses involving PSI factor leads to considerable enhancement of IF; cytoductants with the nucleus of the recipient [psi-] strain not only got [PSI+] factor from the donor strain but also increased their amyloid content. In these model experiments all treatments modifying one of the yeast prions, [PSI+] factor, led to a predictable shift of IF of thioflavin T that behaved like a cytoplasmic hereditary determinant. The data obtained show that IF of thioflavin T staining gives reliable estimates of cellular amyloid content and that mitotically stable shift of IF after a battery of treatments modifying cellular prion set provides quantitative estimate of the input of prionizable protein molecules to the amyloid pool. The combination of thioflavin staining and prionotropic treatments applied here can be possibly used for future attempts of checking yeast strains for cryptic prions. PMID- 18409368 TI - [The microtubule system in endothelial barrier dysfunction: disassembly of peripheral microtubules and microtubules reorganization in internal cytoplasm]. AB - Endothelial cell barrier dysfunction is often associated with dramatic cytoskeletal reorganization, activation of actomyosin contraction and finally gap formation. At present time the role of microtubules in endothelial cell barrier regulation is not fully understood, however a number of observations allow to assume that microtubules reaction is the extremely important part in development of endothelial dysfunction. These observations have been forced us to examine the role of microtubule system reorganization in endothelial cell barrier regulation. In quiescent endothelial cells microtubule density is the highest in the centrosome region and insignificant near the cell margin. The analysis of microtubules distribution after specific antibodies staining using the method of measurement of their fluorescence intensity has shown that in control endothelial cells the reduction of fluorescence intensity from the cell center to its periphery is described by the equation of an exponential regression. The hormone agent, thrombin (25 nM), causes rapid increase of endothelial cell barrier permeability accompanied by fast decrease in quantity of peripheral microtubules and reorganization of microtubule system in internal cytoplasm of endothelial cells (the decrease of fluorescence intensity is described by the equation of linear regress already through 10 min after the beginning of the treatment). Both effects are reversible -- through 60 min after the beginning of the treatment the microtubule network does not differ from normal one, so the microtubule system is capable to adapt for influence of a natural regulator thrombin. The microtubules reaction develops more quickly, than reorganization of the actin filaments system, which responsible for the subsequent changes in the cell shape during barrier dysfunction. Apparently, the microtubules are the first part in a circuit of the reactions leading to the pulmonary endothelial cell barrier compromise. PMID- 18409369 TI - [The study of DNA integrity in somatic cells and spermatozoa by single cell gel electrophoresis with silver staining]. AB - In present work we studied DNA damage in human and bovine lymphocytes and spermatozoa by means of single cell gel electrophoresis followed by silver staining. The spontaneous frequency of DNA damage estimated manually in spermatozoa from healthy donors did not exceed 9% (on average -- 4.8 +/- 1.2%). The frequency of DNA damages in bull sperm after short (less than a year) and long period (more than 20 years) of cryopreservation was assessed as 3.1 +/- 0.9 and 4.3 +/- 0.5%, correspondingly. The comparative estimation of DNA damages in lymphocytes followed by silver staining is a valuable tool to estimate DNA damage in populations of somatic and reproductive cells. PMID- 18409370 TI - [Immunofluorescent analysis of meiotic recombination and interference in the domestic cat]. AB - The aim of this work was an analysis of frequency, density and distribution of recombination sites in male meiosis of the domestic cat. The study was carried out using immunofluorescent staining of synaptonemal complex (SC) proteins, centromeric proteins and mismatch repair protein MLH1, a reliable marker of the sites of crossing over. We mapped 2633 sites of crossing over at 1098 individual autosomes. On the basis of these data the total length of the domestic cat genetic map was estimated as 2176 centimorgans. We found a typical for all mammals studied positive correlation between the length of SC and the number of recombination sites. The domestic cat demonstrated the highest among mammals density of recombination and the lowest interference. PMID- 18409371 TI - [Cytokine-mediated apoptosis of granulocytes eosinophilicus in eosinophilia]. AB - The influence of recombinant forms of cytokincs (IL-5, IL-3 and eotaxin) on programmed destruction of eosinophils obtained from patients with diseases associated with high eosinophilia of blood (malignant diseases of system of blood, opisthorchosis) was studied in vitro. It was shown that all examined categories of patients irrespective of their nosology demonstrated a low level of spontaneous apoptosis of eosinophils. Cultivation of cosinophils isolated from peripheral blood of the patients with invasion O. felineus in vitro with r-IL-5, r-IL-3 and r-eotaxin decreased the number of eosinophilic cells undergoing apoptosis in comparison with spontaneous one. At the same time, incubation of eosinophils obtained from the patients with malignant diseases of system of blood, associated with cosinophilic syndrome, with r-IL-5, r-IL-3 and r-eotaxin allowed to ascertain the absence of sensitivity of eosinophilic cells to the antiapoptotic effect of these cytocines. PMID- 18409372 TI - [Recognition and lysis by natural killers of tumor cells with participation of laminin]. AB - According to the data obtained in the present work, the receptor complex of mouse natural killers (NK) includes laminin, antibody to which blocks EK-activity (NKA regardless of the presence of complement. Preincubation of mouse splenocytes with anti-laminin serum led to a decrease in their NKA towards tumor cells-targets (CT), the NKA activity decreasing 2 times with respect to cultivated cells of rat hepatoma HTC, while 10 times - to cultivated cells of human erythroblastosis K562. Pretreatment of aplenocytes with noraml nonimmune serum did not lead to a change of NKA. Quite different was the pattern after the tumor cell preincubation with anti-laminin serum: pretreatment of CT K562 led to a twofold decrease in sensitivity of these cells to NK-lysis, whereas the pretreatment of CT K562, on the contrary, made them twice sensitive to NK-lysis. Electrophoretic separation of protein of CT plasma membranes with subsequent immunoblotting with anti laminin immune serum revealed the presence oflaminin on HTC cell plasma membrane, which was identified as laminin 8/9 by the mass-spectrometry method, while no laminin was detected on K562 cells. Preincubation of splenocytes with laminin did nor affect NKA with respect to CT K562 and HTC. Pretreatment of CT K562 and HTC with laminin decreased the NKA to zero. The obtained data allow suggesting a doubtless participation of laminin and its receptors in CT cytolysis by NK. PMID- 18409373 TI - [Micronuclear and antitumor activities of two newly synthesized pyrimidine derivatives]. AB - Micronuclear and antitumor activities of two newly synthesized iodides of pyrazolo[1,5a]pyrimidines were investigated using mice model. It has been shown that both compounds are slightly toxic and have no antitumor activity in mice with Ehrlich's ascites carcinoma. Only one of them has weak micronuclear activity. Both compounds increase substantially micronuclear as well as antitumor activities of cyclophosphamide. PMID- 18409374 TI - [The study of non-organic substances content in Henneguya oviperda (Cohn, 1895) and Myxobolus pseudodispar (Gorbunova, 1936) myxospore valves]. AB - The analysis of organic and non-organic substances in the valves of long stored in water Henneguya oviperda (Gohn, 1895) and Myxobolus pseudodispar (Gorbunova, 1936) myxospores has been made using the cytochemical methods and atomic absorptional spectroscopy. The high content of Si (up to 94 %) and the presence of Ca, Mg, Fe, Mn cations (Cu, Al, Ni, Cr in fewer amounts) have been detected. PMID- 18409375 TI - [MADS-box genes controlling inflorescence morphogenesis in sunflower]. AB - MADS-box genes play an important role in plant ontogeny, particularly, in the regulation of floral organ induction and development. Eight full-length cDNAs of HAM (Helianthus annuus MADS) genes have been isolated from sunflower. They encode MADS-box transcription factors expressed in inflorescence tissues. In the frames of the ABCDE model, the HAM proteins were classified according to their structural homology to known MADS-box transcription factors. The HAM45 and HAM59 genes encode the homeotic C function and are involved in the control of the identity of pistil and stamens, while the HAM75 and HAM92 genes determine the A identity of floral and inflorescence meristems and petal identity. The HAM31. HAM2, HAM63, and HAM91 genes encode the B function and are involved in the formation of petals and stamens; and the HAM137 gene encodes the E function. Analysis of the expression of MADS-box genes in sunflower has demonstrated that the structural and functional differences between the ray and tubular flowers in the inflorescence could be a consequence of the lack of HAM59 expression during ray flower initiation. PMID- 18409377 TI - [Cell elongation as an inseparable component of growth in terrestrial plants]. AB - The review is dedicated to the role of cell elongation in plant growth and morphogenesis. The ratios of cell division to elongation, cell competence for the initiation of elongation, main features of the metabolism of elongating cells, and physiological processes realizing elongation have been considered on the examples of seed germination and growth of roots, stems, and leaves. A special attention was paid to the vacuole as a specific feature of plant cells, pathways of its formation, and its role in maintenance of ion and water homeostasis in the elongating cell. The plant can modify its morphology according to changes in the environmental conditions via cell elongation. PMID- 18409376 TI - [Expression of new mutant alleles of AS1 and AS2 genes controlling leaf morphogenesis in Arabidopsis thaliana]. AB - We have studied the morphology and vein branching of rosette leaves in Arabidopsis thaliana mutants as and sa, which proved to be alleles of the A. thaliana AS1 and AS2 genes, respectively. We have also analyzed the localization of bioactive auxin, as measured by the expression of the DR5::GUS transgene, as well as the expression patterns of BP, as measured by the expression of the BP::GUS transgene in leaves of the mutants. In mature leaves of the mutants, BP was expressed ectopically. Furthermore, the mutants showed some defects in the localization and concentration of free auxin compared to the wild type. Our results of studying new alleles of AS1 and AS2 support their role in control of class I KNOX genes and auxin transport. PMID- 18409378 TI - [High molecular weight protein detected in higher plant cells by antibodies against dynein is associated with vesicular organelles including Golgi apparatus]. AB - The cytoplasmic dynein is a multisubunit complex driving organelles along microtubules to their minus-end. We used antibodies against two functional domains (motor and microtubule-binding) of one of principal components of the complex--dynein heavy chain of slime mould Dictyostelium discoideum--to test root meristem cells of wheat Triticum aestivum. The antibodies reacted with a high molecular weight protein (> 500 kDa) in the total cell extract and the band recognized by the antibodies in plant extracts had a lower electrophoretic mobility than the high molecular weight band of mammalian dynein. Antibodies coupled to protein A-Sepharose precipitated the high molecular weight protein from the purified cell extracts. Immunocytochemical analysis demonstrated that the antigen recognized by antibodies against dynein heavy chains is associated with the vesicles whose localization depends on the cell cycle stage. The antigen positive vesicles were localized to the perinuclear region in interphase and early prophase, to the spindle periphery and to spindle pole region during mitosis, and to the interzonal region in the period of fragmoplast and cell plate formation. Some antigen-positive vesicles also reacted with antibodies against Golgi protein markers. The obtained data indicate that higher plant cells contain a high molecular weight protein interacting with antibodies against the motor and microtubules-binding domains of Dictyostelium dynein heavy chain. The revealed antigen was associated with the vesicular structures in the cytoplasm including the Golgi apparatus. PMID- 18409379 TI - [Aquaporins in gametogenesis of vertebrate animals]. AB - A review of the data on the presence, localization, and supposed role of aquaporin water channels in oocytes of Xenopus laevis, oogenesis and maturation of teleosts Sparus auratus and Oncorhynchus mykiss, oogenesis and oocyte maturation of rats and mice, and spermatogenesis of several mammalians. PMID- 18409380 TI - [Analysis of germ cell populations in ejaculate of men infected with herpes simplex virus]. AB - Cytological and molecular genetics methods were used to study sperm from patients with sperm infected with herpes simplex virus (HSV) as indicated by virological and immunocytochemical tests. The following methods were used: (1) sperm analysis to evaluate the morphology and functional properties of sperm; (2) fluorescence in situ hybridization (FISH) with DNA probes specific for chromosomes 1, X, and Y to evaluate nondisjunction frequencies of these chromosomes in sperm; and (3) quantitative analysis of immature germ cells in the ejaculate to identify spermatogenic abnormalities. The total sperm count and the count of sperm with normal motility proved similar to the norm. FISH analysis demonstrated no difference in the nondisjunction frequency of chromosomes 1, X, and Y between infertile patients with HSV-infected sperm and fertile donors. Comparative quantitative analysis of immature germ cells from the ejaculate has demonstrated a significant and considerable (threefold) increase in the number of spermatocytes I at the prepachytene stages of prophase I (preleptotene, leptotene, and zygotene) in HSV patients compared to normal donors. At the same time, HSV patients demonstrated a significant decrease in the number of spermatocytes I, a decrease in the proportion of spermatocytes II and spermatids, and a twofold increase in the number of unidentifiable immature germ cells. The data obtained indicate a partial spermatogenic arrest at the early stages of meiotic prophase I in HSV patients, which prompts further research into the cellular mechanisms of abnormal spermatogenesis after viral infection in humans. PMID- 18409381 TI - [Gonadogenesis and color characteristics of ovaries in Japanese mitten crab Eriocheir japonicus]. AB - The gonadogenesis was studied in adult and juvenile females of Japanese mitten crab Eriocheir japonicus (Crustacea: Decapoda, Grapsida) inhabiting the rivers of the Maritime Territory. The morphometric parameters of oocytes at different stages of maturity were determined using the methods of computer morphometry and color characteristics were evaluated using the Munsell Book of Color. As a result, a color table was compiled for the ovaries from the beginning of development to gonad maturity, which included light yellow (sandy), yellow, beige, light purple, light brown, dark purple, brown (chocolate), and dark brown (brown). The regular changes in the ovary color of E. japonicus proved to closely correlate with the gonadogenesis, namely, with the composition of cells at each stage of gonad maturity. PMID- 18409382 TI - [N-arachidonoyl dopamine is a possible factor of the rate of tentacle formation in freshwater hydra]. AB - The effect of N-arachidonoyl dopamine, haloperidol, and their mixture on the rate of tentacle formation was studied during regeneration of the gastral and basal fragments of freshwater hydra. Some concentrations of haloperidol inhibited the tentacle formation, which was more pronounced in the basal fragment. N arachidonoyl dopamine accelerated the tentacle formation in both fragments, particularly, in the basal one (an inversion of the natural difference in the rate of tentacle formation between the gastral and basal fragments). After the exposure to the mixture of these drugs, the effects of each of them were observed. Mass spectrometry assay has demonstrated endogenous N-arachidonoyl dopamine in the intact hydra homogenate. The possible involvement of this acyl neurotransmitter in the regulation of the rate of tentacle formation in regenerating hydra is discussed. PMID- 18409383 TI - [Macro- and microevolution of bacteria in symbiotic systems]. AB - Using the examples of diverse interactions among prokaryotes and eukaryotes, the relationships between molecular and population mechanisms of evolution of symbiotic bacteria are addressed. Their circulation in host-environment systems activates microevolutionary factors that direct combinative or reductive genome evolution in facultative, ecologically obligatory, and genetically obligatory symbioses. Due to intense systemic intra-genome rearrangements and horizontal gene transfer, two types of gene systems evolve in these bacteria: (1) controlling the pathogenesis-like processes of host recognition and penetration and (2) responsible for mutualistic interactions that are related to nitrogen fixation and its transfer to the host. The evolution of gene systems of type 1 is directed by individual (Darwinian, frequency-dependent) selection, which is responsible for gene-for-gene interactions between the partners. In the evolution of the type 2 systems, group (interdeme, kin) selection plays the key role, being responsible for the development of bacterial traits beneficial for the host. Using the legume--rhizobia symbiosis as an example, it is shown that evolution of mutualism can be described in terms of biological altruism, whose regularities are common for intraspecific and interspecific relationships. Macroevolutionary rearrangements of bacterial genomes result from the structural changes in their populations, wherein various selection modes are combined with stochastic processes (genetic drift, population waves) induced in the symbiotic systems. PMID- 18409384 TI - [Quasi-adaptive response to alkylating agents in Escherichia coli and Ada-protein functions]. AB - In 2005 we have described in exponentially growing E. coli cells a new fundamental genetic phenomenon,--quasi-adaptive response to alkylating compounds (quasi-Ada). Phenotypic expression of quasi-Ada is similar to the true Ada response. However, in contrast to the letter, it develops in the course of pretreatment of the cells by a sublethal dose of nonalkylating agent, an NO containing dinitrosyl iron complex with glutathione (DNICglu). To reveal the mechanisms of quasi-adaptation and its association with the function of the Ada regulatory protein, here we used a unique property of dual gene expression regulation of aidB1 gene, a part of the Ada-regulon, namely its relative independence from Ada protein in anaerobic conditions. Based on the results of aidB1 gene expression analysis an EPR spectra of E. coli MV2176 cells (aidB1::lacZ) in aerobic and anaerobic conditions after the corresponding treatments, we conclude that the function and the spatial structure of meAda and [(Cys-)2Fe+(NO+)2]Ada are identical and thus the nitrosylated protein represents a regulator of the Ada regulon gene expression during quasi-adaptation development. PMID- 18409385 TI - [Isolation of antibiotic resistance bacterial strains from East Siberia permafrost sediments]. AB - A collection of bacterial antibiotic resistance strains isolated from arctic permafrost subsoil sediments of various age and genesis was created. The collection included approximately 100 strains of Gram-positive (Firmicutes, Arthrobacter) and Gram-negative bacteria (Bacteroidetes, gamma-Proteobacteria, and alpha-Proteobacteria) resistant to aminoglycoside antibiotics (gentamycin, kanamycin, and streptomycin), chloramphenicol and tetracycline. Antibiotic resistance spectra were shown to differ in Gram-positive and Gram-negative bacteria. Multidrug resistance strains were found for the first time in ancient bacteria. In studies of the molecular nature of determinants for streptomycin resistance, determinants of the two types were detected: strA-strB genes coding for aminoglycoside phosphotransferases and genes aadA encoding aminoglycoside adenylyltransferases. These genes proved to be highly homologous to those of contemporary bacteria. PMID- 18409386 TI - [Genetic polymorphism of Siberian spruce (Picea obovata Ledeb) in middle Siberia]. AB - Based on the analysis of 22 loci controlling allozyme variation of MDH, SKDH, 6 PGD, IDH, PEPCA, GOT, FDH, LAP, PGI, PGM, SOD, and GDH, the data on within- and among-population variation for nine cenopopulations of Siberian spruce (Picea obovata Ledeb.) located along the Yenisei meridian, from 65 degrees 50' NL to 52degrees 14' NL, were obtained. It was demonstrated that 86.36% of the loci, tested in the species analyzed, were polymorphic. The mean number of alleles per locus was 2.91, and the observed and expected heterozygosity constituted 0.161 and 0.168, respectively. More than 97% of total variation occurred within the populations, while the proportion of the among-population variation constituted only 2.3% (Fst =0.0230). Genetic distance (DN) between the populations examined varied from 0.0019 to 0.0115, averaging at 0.0051. It was shown that in the part of the Siberian spruce range examined there was no close association between the level of the genetic differentiation of the populations and the geographic distance between them. It seems likely, that this finding can be associated with the fact that Siberian spruce, growing on this territory, is the intrazonal species, i.e., it is not attached to a certain forest zone, and its distribution is mostly determined by local ecological conditions. PMID- 18409387 TI - [Recombinant genome of cereals: the pattern of formation and the role in evolution of polyploid species]. AB - The pivotal-differential model of evolution of polyploid species of cereals has been experimentally reproduced, and the pattern of the formation of a recombinant genome has been analyzed. It has been found that mutual substitution of chromosomes of the original genomes is subjected to selection pressure and, hence, is nonrandom. The selection occurs at the level of homeologs, whose selective advantages are determined by interactions between the genotype and the environment. If a homeolog has distinct selective advantages, the chromosomal composition of the corresponding homeologous group is completed rapidly, which leads to the formation of intergenomic recombination at the level of whole chromosomes. If homeologs have the same competitiveness, the composition of the group is stabilized more slowly. Domination of the genetic systems of the basic genome ensures a high rate of pairing of homeologous chromosomes of the recombinant genome during meiosis, which leads to recombinations at the level of chromosomal segments. It has been demonstrated that different combinations of chromosomes from original genomes are selected at different conditions of plant growth. PMID- 18409388 TI - [Genealogical analysis of the diversity of spring barley cultivars released in former Czechoslovakia and modern Czechia]. AB - Genealogical analysis was employed in studying the time course of changes in genetic diversity of spring barley cultivars released in former Czechoslovakia and the modem Czech Republic. Cultivars from different regions proved to significantly differ in the distribution of dominant ancestor contributions, suggesting a specificity of original ancestors to different cultivation conditions. A comparison of cultivar groups differing in end use showed that the genetic diversity of malting cultivars was significantly lower than that of feed cultivars, although modern malting and feed cultivars of Czechia and Slovakia have virtually the same genetic basis. Temporal analysis showed that diversity tended to increase through decades. While new original ancestors were introduced in pedigrees, especially in the past 30 years, the number of local landraces and old cultivars gradually decreased. The losses accounted for about two-thirds of the local germplasm. Thus, a substantial increase in genetic diversity was accompanied by genetic erosion of the local spring barley gene pool of former Czechoslovakia. A cluster structure was observed for the set of spring barley cultivars released in the postwar period. The coefficient of parentage averaged overall possible pairs of cultivars introduced in the Czech National List was estimated at 0.11. It was concluded that the genetic diversity of modern spring barley cultivars in the Czech Republic is at an acceptable level. PMID- 18409389 TI - [Analysis of serial sequences of seeds in sugar beet plants upon apozygotic reproduction]. AB - Dichotomic serial sequences of fruits of two types (with and without seeds) on shoots were analyzed in sugar beet upon apozygotic reproduction. Serial sequences were analyzed using serial statistical tests. The distribution of fruits of the two types was nonrandom in most cases. The variation of serial sequences is considered in terms of the epigenetic variation of discrete characters in plants. PMID- 18409390 TI - [Construction and molecular and cytogenetic analyses of euploid (2n = 42) and telocentric addition (2n = 42 + 2t) alloplasmic lines (Hordeum marinum subsp gussoneanum)-Triticum aestivum]. AB - Individual plants from the BC1F5 and BC1F6 backcross progenies of barley--wheat (= H. geniculatum All.) (2n = 28) x T. aestivum L. (2n = 42)] and the BC1F6 progeny of their amphiploids were used to obtain alloplasmic euploid (2n = 42) lines L-28, L-29, and L-49 and alloplasmic telocentric addition (2n = 42 + 2t) lines L-37, L-38, and L-50. The lines were examined by genomic in situ hybridization (GISH), microsatellite analysis, chromosome C-banding, and PCR analysis of the mitochondrial 18S/5S repeat. Lines L-29 and L-49 were characterized by substitution of wild barley chromosome 7H1 for common wheat chromosome 7D. In line L-49, common wheat chromosomes 1B, 5D, and 7D were substituted with homeologous barley chromosomes. Lines L-37, L-38, and L-50 each contained a pair of telocentric chromosomes, which corresponded to barley chromosome arm 7H'L. All lines displayed heteroplasmy for the mitochondrial 18S/5S locus; i.e., both barley and wheat sequences were found. PMID- 18409391 TI - [Mapping of the quantitative trait loci (QTL) associated with grain quality characteristics of the common wheat grown under different environmental conditions]. AB - The quantitative trait loci (QTL) associated with individual characteristics of grain and flour quality in wheat lines grown under contrasting environmental conditions were mapped. Overall, 22 QTL that manifested under contrasting environmental conditions with various significances were detected on 10 chromosomes. Grain hardness and vitreousness were associated with three loci on chromosomes 5D, 6A, and 3A, while the gluten content, with two loci on chromosomes 5B and 7A. Dough extensibility was associated with only one QTL localized in the region of Glu-A1 locus. One of the loci determining flour and dough strengths is located in the region of Gli-B1 and Glu-B3 loci and the rest, in various regions of chromosomes 1B, 5D, and 4B, where no particular genes associated with grain quality have been yet found. The detected QTL can be used in further experiments on genetic control of gluten formation and quality in wheat. PMID- 18409392 TI - [Genetic regulation of the centromere division in rye and wheat univalent chromosomes in dimonosomics during meiotic anaphase I]. AB - Cytogenetic analysis of meiosis in the wheat--rye dimonosomics 1Rv-1A, 1Ron-1A, 2R-2D, 5R-5A, and 6R-6A was conducted. C-banding was used to study the segregation pattern of each of two univalent chromosomes during the first meiotic division. It has been shown that the division frequency of the centromeric regions of all rye chromosomes in the pair studied is significantly higher than in the wheat chromosomes. The ANOVA performed suggest that the plant genotype contributes significantly (at P = 0.05) to the behavior pattern of univalent chromosomes in meiosis. The data obtained demonstrate that the rye and wheat chromosomes studied are involved in genetic regulation of centromere division in meiotic anaphase I (AI). The presence of rye chromosome 2R and wheat chromosome 2D suppresses the division of centromeres of the sister chromatids in AI. Rye chromosomes 1Rv, 1Ron, 5R, and 6R induce equational division; however, rye chromosome 1Rv increases to a greater degree the frequency of equational division of wheat chromosome 1A as compared with chromosome 1Ron. PMID- 18409393 TI - [Restriction fragment variation of the mitochondrial gene of cytochrome b in some taxa of the Eelpout family (pisces, perciformes, zoarcidae) from the northern Sea of Okhotsk]. AB - The first analysis of restriction fragment length polymorphism (RFLP) of the mitochondrial gene of cytochrome b of fish from the Eelpout Family (Zoarcidae) from the northern Sea of Okhotsk has been performed. The mean genetic distances between the subfamilies Zoarcinae and Lycodinae, subfamilies Lycodinae and Gymnelinae, and subfamilies Zoarcinae and Gymnelinae are 11.14, 13.87, and 14%, respectively. Species of the genus Lycodes (Lycodinae) are characterized by a mtDNA small divergence (on average, 2.04%) and are divided into two groups on the basis of lateral line morphology. Magadania skopetsi, an endemic of Tauiskaya Bay of the Sea of Okhotsk, is genetically close to Hadropareia middendorffii from the same subfamily (Gymnelinae). The mtDNA sequences of species from the genus Lycogrammoides (Lycodinae) differ by 3.83%. The genetic differences between the eelpout Zoarces elongatus Zoarces sp. (d = 4.54%) suggest that the latter is a separate species. PMID- 18409394 TI - [Classification analysis of a latent dinucleotide periodicity of plant genomes]. AB - The information decomposition (ID) method has been used for searching dinucleotide periodicities, including latent ones, in plant genomes. In nucleotide sequences of genomes of various plants from the GenBank database, 14766 sequences with a periodicity of two nucleotides have been found. Classification of the periodicity matrices of the detected DNA sequences has yielded 141 classes of dinucleotide periodicity. Since ID does not detect periodicities with nucleotide deletions or insertions, modified profile analysis (MPA) has been applied to the obtained classes to reveal DNA sequences with dinucleotide periodicities containing nucleotide deletions and insertions. Combined use of ID and MPA has permitted the detection of 80 396 DNA sequences with dinucleotide periodicities in the genomes of various plants. The biological role of dinucleotide periodicity in the detected sequences is discussed. PMID- 18409395 TI - [Molecular analysis of the phylogenetic relationships among the diploid aegilops species of the section Sitopsis]. AB - RAPD analysis was used to study the intraspecific variation and phylogenetic relationships of S-genome diploid Aegilops species regarded as potential donors of the B genome of cultivated wheat. In total, 21 DNA specimens from six S-genome diploid species were examined. On a dendrogram, Ae. speltoides and Ae. aucheri formed the most isolated cluster. Among the other species, Ae. searsii was the most distant while Ae. longissima and Ae. sharonensis were the closest species. The maximum difference between individual accessions within one species was approximately the same (0.18-0.22) in Ae. bicornis, Ae. longissima. Ae. sharonensis, and Ae. searsii. The difference between the clusters of questionable species Ae. speltoides and Ae. aucheri corresponded to the intraspecific level; the difference between closely related Ae. longissima and Ae. sharonensis corresponded to the interspecific level. The section Sitopsis of the genus Aegilops includes six diploid species containing the S genome, which is regarded as an ancestor of the B genome of cultivated wheat. The species of the section are thought to be closest to the genus Triticum. Note that the taxonomic status of some forms of the section Sitopsis is questionable. For instance, Ae. speltoides and Ae. aucheri are variously considered as individual species or as a single species, Ae. speltoides. The situation with Ae. longissima and Ae. sharonensis is similar. Thus, although the group includes only diploid species and is well studied morphologically, its phylogeny and taxonomy are still questionable. PMID- 18409396 TI - [Phenomenological models of the forest plantations growth]. AB - Phenomenological models of the forest plantations growth are analyzed. Those derived from the Verhulst's model are shown to fail describing qualitative effects reflecting tree growth, phytomass withdrawals, and plantation restoration. The method of phase portraits is used for exploration of the forest ecosystem dynamics, which allows to describe regulatory mechanisms of the growth processes, regulation delay, and feedback types. A bistable phenomenological model is suggested herewith to characterize dynamic processes in the forest ecosystems. Principal patterns of formation of the forest plantations at different stages of the forest generation processes are considered on the basis of that model, and ecological effects responsible for the plantation dynamics are revealed. PMID- 18409397 TI - [Fungi: individuals, populations, and speciation]. AB - Under considerations are specific features of the fungal individuals associated with their mycelial life mode and affecting their population structure and speciation. Special attention is paid to the sympatric speciation which can be caused by trophic niche segregation, by use of different host plants, by invasion to the plants at different stage of their ontogeny, and by differences in the weather requirements. Decrease of genetic interchanges in subdivided populations promoting speciation without spatial isolation is achieved by homotallism and pseudohomotallism, by a cassette mechanism of switching among various breeding systems, and by complete lost of sexual process. Recombination reduction in agamic fungi is achieved by means of vegetative incompatibility. As the latter is similar functionally to the immune system (recognition of an alien culture and death of conjugated cells), it is possible that the fungi were the first to have developed mechanisms of sympatric speciation on the basis of the simpliest immune system. PMID- 18409398 TI - [How and why the mechanisms of ageing and carcinogenesis originated: a hypothesis]. AB - A hypothesis explaning the nature of ageing and carcinogenesis is proposed. The hypothesis is based on the notion of a unitary organism as a module of a colony that has lost the ability to reproduce asexually. The evolutionary forerunner of the old age involution was the asexual iteroparity of colony modules, and the evolutionary forerunner of carcinogenesis was the semelparity of such colonies; the evolutionary forerunner or tumour regression was the switch of the colony module from semelparity to iteroparity. The donor-acceptor relations of a colony module emerge in unitary organisms in the course of ageing and carcinogenesis. The mechanisms of ageing and carcinogenesis originated at the stages of Metazoa evolution when the organisms had not yet acquired the nervous and the endocrine systems. PMID- 18409399 TI - [Geographic variation as a result of evolution of the traits with broad and narrow norms of reaction in the moor frog (Rana arvalis)]. AB - Females reproductive, size, and age characteristics were studied in isolated local populations of Rana arvalis in the southern and northern parts of its range. The yearlings of the southern populations used to get larger by their first overwintering due to earlier beginning of the breeding season, as compared with the yearlings of the northern population. As a result, "southern" females become sexually mature at the age of two years while the "northern" ones become mature at the age of three years. This causes geographic differences in age composition among two populations, the "southern" reproductive females being younger on average than the "northern" ones. The earlier female maturation in the first case is not compensated by respective rise of the growth rate; to the contrary, the "southern" females grow more slowly during the first two years of their life and appear to be smaller than the "norhern" ones. These reproduction and growth patterns arise supposedly due to paedomorphosis, which causes specific reproductive characteristics, namely decrease in the egg size, increase in the reproductive effort and more strong correlation between female fertility and body size. Local and geographic differences are expressed not in the extent but in the structure of reproductive pattern, as no negative correlation was revealed between female reproductive age and body size in the southern populations. Southern habitats cannot be considered as "unfavourable with respect to body size", so the geographic differences under consideration cannot be explained by optimization of the reproductive strategies at population level. Paedomorphosis appears as a result of the female maturation rate possessing a wider norm of reaction than the growth rate. At the same time, fixation of the specific growth rate narrows norm of reaction of some other characters important for the phenotype reproductive fitness thus predetermining their subsequent evolution. PMID- 18409400 TI - [The notion of ecological optimum and the determination of this optimum in freshwater poikilothermic animals]. AB - The notion of ecological optimum is discussed as related to aquatic poikilothermic animals. Publications on the analysis of the influence of thermal factor on vital functions of invertebrates with the subsequent statement of the general pattern of optimal environmental factors are reviewed. It is concluded that the optimum includes not simply the range of the factor's values on the tolerance curve or the dose of the factor providing for the most favourable life conditions, but the oscillations of the factor within the optimum range, i.e., the astatic optimum, as termed by Prof. A.S. Konstantinov. It is shown, by the example of the influence on organisms and populations of zooplankton of stepwise changes of temperature, increased doses of mineral phosphorus, and low-power laser radiation, that to determine the optimum of an environmental factor it is important to use not only its absolute values and the parameters of cyclic changes, but also the presence or absence of stepwise changes, also determining the direction of the dynamics and the duration of influence of the "dose", as well as the presence and character of the after-effects (inertial action) of the factor on organisms. It is suggested to introduce more detailed and concrete definitions to distinguish between the "static" and then "dynamic" optimum. The former includes the range of optimum values of the factor on the tolerance scale and the "dose" of every factor matching the organism's requirements and providing for the maximum favourable life conditions. The latter includes optimum parameters of cyclic changes (frequency and amplitude) of the factor, along with determining their position in the range of optimum values (at its lower, middle, or upper part), the presence or absence of stimulating effects of stepwise changes of the factor, including the duration of influence of some "dose" of the factor and the order of alteration of the higher and the lower values (dynamics direction), as well as the presence or absence and the character of after-effects (inertial action) of the factor on the organism. PMID- 18409401 TI - [Degradation succession of heterotrophic flagellate communities in microcosms]. AB - Degradation succession of heterotrophic flagellate communities was studied in samples of detritus and water from a small swamped lake in the environs of the settlement Borok, Yaroslavl province. Three stages of succession with different species composition of the heterotrophic flagellate community were clearly recognized. Cumulative properties of the community change with time. Maximum species richness, diversity, and abundance were observed on the fourth day of succession; their values gradually decreased later. The trophic and taxonomic community structure was most diverse at the earlier stages of succession. Primordial food (bacteria) concentration affects only the size of peak community properties, but has no influence on the pattern of succession changes. The influence of predators considerably reduced heterotrophic flagellate abundance and can shift the time of the stage with maximum species diversity in the course of succession. PMID- 18409402 TI - [Morpha striata in the members of the genus Rana (Amphibia, Anura), the reasons of adaptability to environmental changes]. AB - Under investigation is a complex of inherited physiological properties of the morpha striata (a monogenous dominant mutation) in two species of the genus Rana. Insufficient effectiveness of the potassium-sodium pump responsible for the skin transport in amphibians had lead to formation of a number of compensative physiological mechanisms in this morpha. The yearlings of the morpha striata are characterized by highly dynamic hemopoetic system playing important role in individual adaptations to unstable environments. Such a high level of metabolism in the morpha striata promotes rising of adaptive potential of the nervous system due to decrease of the excitability threshold, but causes shortening the life span. Therefore, physiological differences correlated with polymorph structure of the close species can be of crucial importance in their adaptations under existence in the natural and artificial geochemical anomalies and in anthropogenically disturbed ecosystems. PMID- 18409403 TI - [Comporative characteristics of action potentials in animals and higher plants]. AB - The current comparative characteristics of action potentials in animals and higher plants are given. The main differences in parameters, mechanisms of generation, and functional significance of propagated electrical signals are examined. PMID- 18409404 TI - [MRI of fetal central nervous system malformations]. AB - Ultrasonography is the method of choice for prenatal malformation screening, but it does not always provide sufficient information for correct diagnosis or adequate abnormality evaluation. Fetal magnetic resonance imaging (MRI) is considered a valuable second line imaging tool for confirmation, completion and correction of complex fetal ultrasonography findings. Fetal MRI has proven its value in pathologies of the central nervous system. MR imaging's role includes nor only confirming or excluding possible lesions, but also defining their full extent, aiding in their characterization, and demonstrating other associated abnormalities. The article describes the most common anomalies of CNS discover by fetal MR. PMID- 18409405 TI - [MRI brain imaging data in children with cavum septi pellucidi and vergae]. AB - BACKGROUND: Cavum septum pellucidum (CSP) is commonly known marker of neurodevelopmental brain malformation. In MR imaging it is seen either as an isolated finding or with association of others developmental brain changes e.g. cavum vergae (CV). AIM OF THE STUDY: was to assess the frequency of CSP and CV associated with others brain developmental malformations in children. We also classified clinical syndromes in children with these structural brain malformations. MATERIAL AND METHODS: The study was performed on 55 children, hospitalised in the Department of Pediatric Neurology, Jagiellonian University in Krakow, between 1998-2006 with diagnosed CSP and/or CV in MRI. There were 29 girls and 26 boys aged 4 months-16 years old. Clinical history, clinical status and neurological findings in physical examination were main indication to perform brain MRI. MRI scans was done in SE T1, FSE T2, PD, FLAIR, IR T1 sequences, 3 and 5 mm depth, in vertical, frontal and axial surfaces, performed on 1,5 T machine Signa Horizon HiSpeed General Electric. RESULTS: There were 38 children with CSP, 12 children with CSP and CV, and 5 with CV. In 18/55 children (33%) CSP was isolated, but in 37/55 children (67%) CSP was coexisted with other brain structural malformations. The average age of diagnosis in isolated CSP was 10 years old, in coexisting malformation was 5 years. In half of the children, the reason of neuroimaging was epilepsy, in this 6/18 patient with isolated CSP or CV. 58% of the study group had a normal mental development, but 35% of children with additional brain malformation and 11% of children with isolated CSP and CV were mentally retarded. CONCLUSION: 1. The majority of the children with CSP and CV had also coexisting brain malformations diagnosed in MRI. 2. CSP and CV coexisting with large brain malformation e.g. dysplasia septo-optica and agenesis of corpus callosum. were diagnosed earlier. 4. Mental retardation was diagnosed in 1/3 children with different structural brain malformation but also in 11% of children with CSP and CV as well. PMID- 18409407 TI - [MRI and MRA data in children with migraine with aura]. AB - INTRODUCTION: One of indications to perform structural MRI and MRA is migraine with aura. Results may influence migraine management. THE AIM: THE AIM of this study was an assessment of MRI and MRA results in children with symptoms of migraine with aura and also the analysis if MR results influence migraine management. MATERIALS AND METHODS: During the year 2006, 20 children with migraine were hospitalized in the Department of Pediatric Neurology Jagiellonian University Krakow. There were 11 girls and 9 boys, aged 6-18 years (mean 13,7 years) with symptoms of migraine with aura. In 12 of them visual, in 9 sensory, in 6 dysphasic and in 3 motor aura was present. In 7 children two or more aura types coexisted. In 9 cases family history of migraine was positive. MRI was performed in all children, while additional MRA in 18 children and digital cerebral angiography in 1 of them. MRI was performed in SE T1, FSE T2, PD, FLAIR and IR T1 sequences with 3 or 5 millimetre thickness of slices, in the sagital, coronal and transversal plane, using a 1,5 T system (HiSpeed GE). 3D TOF techniques were used to perform MRA. RESULTS: Results of MRI were normal in 11/20. In other children brain pathology was detected, within lateral ventricular asymmetry (2), demyelination (2), dilatation of CSF spaces (2) and vascular malformation (2). Isolated pathology, such as a calcification of the pineal gland and focal demyelination of vascular origin were detected in individual children. MRA in 16 children did not revealed pathology, however in 2 other children slight asymmetry of distal insular branches of middle cerebral arteries was detected. Aneurysm of the right anterior communicans artery was suspected in one child and it was confirmed by MRA. In one case digital angiography confirmed described on MRI result left vertebral and cerebral posterior arteries hypoplasia and also revealed collaterals in the posterior skull cavity. CONCLUSIONS: Detection of vascular pathology in 20 % of patients with migraine with aura using MRI and MRA is an argument to perform both examinations in such patients. PMID- 18409406 TI - [Assessment of the Doppler cerebral blood flow measurement in infants with perinatal trauma]. AB - BACKGROUND: Birth trauma may lead in infants to developmental delay, cerebral palsy, epilepsy and complications with increasing risk of the death. Several complications are of vascular origin. The aim of the study was the assessment of the transcranial Doppler-sonographic (TCD) measurement of cerebral blood flow in infants with perinatal CNS pathology. MATERIAL AND METHODS: 14 infants, 9 females and 5 males, within 12 premature infants, with the CNS perinatal pathology were examined. In children during the neonatal period respiratory distress syndrome, sepsis, apnoe and seizures were also observed. Neuroimaging was performed in each neonate and TCD measurement as well, in 1-7 month of age. Pulsatility index (PI) and resistive index (RI) in anterior cerebral artery (ACA) and in middle cerebral artery (MCA) were established. RESULTS: In 10 children neuroradiologic examination revealed pathological changes: intraventricular hemorrhage, subdural hematoma, subarachnoid hemorrhage, porencephalic cyst of parietal lobes, agenesis of corpus callosum or lateral ventricles dilatation. In 7 children of this group blood flow changes were registered and in 3 of them (21%) the blood flow was normal. The most severe decrease of blood flow in ACA and MCA associated with decreased PI and RI were detected in a case after subdural hematoma operation, prior to death. In other 4 children with normal brain imaging but abnormal neurological examination, cerebral blood flow was disturbed. In three of them blood flow velocity in ACA was decrease with PI increase and in other one blood flow velocity in MCA was increased. CONCLUSIONS: 1. In 79% of the infants with CNS perinatal pathology transcranial Doppler identified blood flow changes. 2. The further research is necessary for the confirmation that a significant decrease of blood flow velocities and indexes in such cases is a poor prognostic factor. PMID- 18409408 TI - [Results of cerebral angiography performed after neuroradiological indications in children with CNS diseases]. AB - INTRODUCTION: Despite of the constant progress in the imaging of vessels, cerebral angiography characterises with the best specificity and sensitivity in detection of arterial, arteriovenous malformations and aneurysms. THE AIM: The aim of this study was to compare the results of cerebral angiography and other neuro-radiologic methods in cases suggesting the probability of vascular malformations in children with neurologic syndroms. MATHERIAL AND MEHTODS: In the year 2006 cerebral angiography was performed in 20 children since it was advised after radiological examinations in children hospitalized in the Department of Pediatric Neurology UJ CM in Krako w. The group of children aged 3-17 years (mean 11 years) consisted of 11 girls and 9 boys. The commonest clinical diagnoses were epilepsy, less common cranial neuropathies, migraine, hemiparesis and agiomatosis of skin. Before cerebral angiography was indicated, other radiological examinations were performed, within CT in 7 cases, CTA in one, MRI in 17 children and MRA in 5. In some children other examinations were also performed, within transcranial doppler evaluation of cerebral blood flow, EEG, video EEG, lumbar puncture and ultrasound eye evaluation. RESULTS: The pathological changes were detected in 5 CT examinations, 1 CTA, 7 MRI and in 4 MRA, suggesting cerebral vascular malformations. Cerebral angiography excluded vascular malformations in 15 children. In 5 children angiography confirmed however the presence of vascular pathology. In 2 of them arterial hypoplasia in the posterior circulation was detected, in 2 cases supratentorial artery hypoplasia and a single venous cerebellar malformation. Children were diagnosed with migraine with aura in two cases, within one with paresis of the right leg and single cases with skin angiomatosis, syncope and sleep myoclonus. CONCLUSIONS: Cerebral angiography allowed to exclude vascular pathology in 75% of children, in whom previous neuroradiological examinations suggested such pathology. It was important for their quality of life. In 25% of patients angiography confirmed results of previous neuroradiological tests detecting vascular pathology. MR with contrast medium is more sensitive than angiography in detection of venous angiomas. PMID- 18409409 TI - [Brain MRI data in children with so called primary generalized seizures]. AB - INTRODUCTION: Diagnosis of primary generalised seizures may be not precise because difficulties exist with the assessment of information of focal onset of these seizures. THE AIM: The aim was to examine if the careful searching for partial component of seizures is necessary in children with so called primary generalized seizures. MATERIAL AND METHODS: The examined group included 45 children aged 6 month to 18 year (mean 11 year) within 19 girls and 26 boys, hospitalized during the year 2006 in the Department of Pediatric Neurology UJ CM in Krakow. The history indicated the primary generalized character of seizures. The tonic-clonic seizures were observed in 39 children, myoclonic in 3, infantile spasms in 1 and absence in other 2. In 6 patients familiar epilepsy was diagnosed in first-degree cousins. Neuropediatric examination was normal apart from 4 children with dysmorphic features of the face. Intellectual development was normal in 43 children, however it was close to the lower range in one and mildly delayed in the other one. Generalized paroxysmal bioelectric activity was detected in 43 children and in 2 children EEG pattern was normal. MRI was performed in SE T1, FSE T2, PD, FLAIR and IR T1 sequences with 3 or 5 millimetre thickness of layer in the sagital, coronal and transversal plane, using a 1.5 T signa horizon HiSpeed General Electric. These examinations were performed in the Voxel MR Laboratory in University Children's Hospital in Krakow. RESULTS: MRI results were normal in 14/45 children and in others pathological. In 9/45 MR revealed subarachnoidal cyst, in 8 lateral ventricles asymmetry, in 5 mild demyelination, in 9 subarachnoid space and lateral ventricles dilatation. In 2/45 children brain tumors (hamartoma of the hypothalamus and DNET of the temporal lobe), in 1 gliosis of the occipital lobe were revealed, however other insignificant changes were also observed. Dual or multiple MRI pathology was revealed in 14 children. In17/45 children abnormalities were localized. CONCLUSIONS: Existence of focal pathology in MRI of 38% children with the history of so called primary generalized seizures suggested the necessity of more careful searching for partial component of seizures. PMID- 18409410 TI - [Results of control MRI examinations performed due to radiological indication- preliminary report]. AB - INTRODUCTION: MR is very important examination of central nervous system in children in the evaluation of differences dependent of sex and age as well as in the diagnostics. Radiological indications are important in many cases to perform a control study, to evaluate a dynamic changes or to establish the diagnosis. THE AIM: The aim of this study was to examine the results of control MRI examination in children with neurologic syndromes. MATERIAL AND METHODS: In the year 2006 control MRI was performed in 22 cases since it was advised after previous MRI examinations in children hospitalized in the Department of Pediatric Neurology UJ CM in Krakow. The group of children aged 4-18 years (mean 12 years) consisted of 14 boys and 8 girls. The commonest clinical diagnoses were epilepsy, encephalopathies, hemiparesis, cranial neuropathies, headache, anizocoria, vertigo, paraparesis and NF I. RESULTS: Repeated MRI in 14 children did not revealed significant changes, independently from clinical changes. In 3 patients positive changes or regres of pathological foci were observed within the subarachnoid hemorrhage. In 3 children MRI revealed progressive changes, within in the child with NFI, and in patient with pyramidolextrapyramidal syndrome which progressed from hemiparesis to quadriplegia and also in a child with disseminated symptoms, such as ataxia, piramidal syndrome and cranial nerve palsies. In 2 other children the repeated MRI helped to establish the final diagnosis. The evolution of neuroradiological changes in the first child with the history of hemiparesis indicated ischemia as a cause of neurological syndrome. The other patient with cranial neuropathies of VII, VIII and XII nerves associated with intellectual impairment was diagnosed with CADASIL only with the help of control MRI examination. CONCLUSIONS: In 9% children the second MRI which helped to analyze evolution of changes was important to establish the final diagnosis. The control MRI examination in 64% children did not revealed significant changes independently of clinical changes, such as recovery or aggravation of symptoms, what explains the decision to repeat MR examination in unclear cases. PMID- 18409411 TI - [Trigeminalgia caused by neurovascular compression in 12 years old girl]. AB - Trigeminalgia is one of the most frequent clinical problems, common in adults but also found in children. In this paper we described a case of 12 years old girl with symptomatic trigeminalgia caused by neurovascular compression, hospitalized in the Department of Pediatric Neurology Jagiellonian University in Krakow. It creates a very difficult diagnostic problem. The girl was first unsuccessfully treated with carbamazepine and afterwards the surgery of neurovascular decompression was performed. We emphasis the crucial role of MR and MRA in cases refractory to classic pharmacotherapy. PMID- 18409412 TI - [The role of the magnetic resonance spectroscopy in the diagnosis of temporal lobe epilepsy]. AB - Epilepsy is the most common neurological disease. Depending on the site of the origin of seizures epilepsy may be divided into the focal and generalised forms. Partial epilepsy most often occurs in the drug-resistant form, the most frequent variant of which is temporal lobe epilepsy--TLE. In the case of focal lesions the removal of the area responsible for the origin of seizures may result in good therapeutic outcome. It is, therefore, necessary to precisely localise an epileptogenic focus. This may be achieved using neuropsychological tests, electroencephalography (EEG), magnetic resonance imaging (MRI), positron emission tomography (PET), single photon emission computed tomography (SPECT), functional magnetic resonance imaging (fMRI), magnetic resonance spectroscopy (MRS) and diffusion magnetic resonance imaging (dMRI). There is a considerable group of patients in whom the cause of the disease is a subtle structural lesion that could not be found using CT or MRI, or a functional cause. In such cases novel techniques are used that show functional causes, e.g. the change in blood perfusion, cellular components concentration or neurotransmitter binding. Those techniques involve: functional MR, PET, SPECT, dMR and MRS. Due to the limitations of other methods magnetic resonance spectroscopy has played an increasing role in the imaging diagnostics of epilepsy. MRS is a non-invasive method of the assessment of the chemical composition of a tissue sample. In persons with epilepsy HMRS of the hippocampus may show the changes in specific metabolites. Several studies have also shown that HMRS is a useful non-invasive method of monitoring the response to anti-epileptic drugs. PMID- 18409413 TI - [The neurochemical mechanisms of temporal lobe epilepsy: an update]. AB - BACKGROUND: Last decade witnessed a remarkable progress in understanding the molecular background of temporal lobe epilepsy. That success was due not only to undisputable achievements in the development of electrochemical and biological techniques, but also to the well-defined neuropathological features of partial epilepsy and the availability of appropriate animal models. THE AIM: The aim of the present paper was to review recent reports concerning those issues. Special emphasis was put on glutamatergic and GABAergic systems, but the putative role of some neuropeptides, ion channels and intracellular signaling pathways in the pathomechanism of temporal epilepsy was also discussed. RESULTS: The most intriguing and still unsolved problem with studying the basic mechanism of partial epilepsy is a mutual relationship between hippocampal damage and epileptogenesis, in particular the interaction of glia and neuronal factors in this long-term dynamic process. CONCLUSIONS: It is pointed out that better understanding of these molecular processes may have clinical implications in the future as regards preventing partial seizures, an accurate diagnosis and obtaining safer and more efficient antiepileptic drugs. PMID- 18409414 TI - [Caffeine and antiepileptic drugs: experimental and clinical data]. AB - BACKGROUND: Caffeine, a methylxanthine derivative, is contained in coffee or tea, chocolate as well as in some beverages. In addition, it may be added to some analgesics. At high doses, similarly to other methylxanthine derivatives (theophylline, pentoxifylline) caffeine induces seizure activity in rodents. THE AIM OF STUDY: If caffeine intake from coffee drinking resulting in pharmacologically active plasma caffeine concentrations--can lead to diverse interactions with other medications. RESULTS: Since 90s of the XX century, there are experimental data available pointing to the caffeine-induced impairment of the protective activity of a number of antiepileptic drugs in basic models of epilepsy in rodents. Acute caffeine, in doses far below its convulsive potential (almost 10-20 fold lower than the ED50 of the methylxanthine of 2.03 mmol/kg for the induction of seizures), produced a significant reduction in the anticonvulsant effects of carbamazepine, phenobarbital, phenytoin, and valproate against maximal electroshock-induced seizures in mice. This interaction was pharmacodynamic in nature since caffeine did not affect the plasma concentrations of these anti-epileptics. Interestingly, there was no tolerance to this hazardous effect of caffeine since its administration at the same dosages (0.12-0.24 mmollkg) also resulted in the impairment of the protection provided by antiepileptic drugs, this effect being even more pronounced in the case of phenobarbital and carbamazepine. In case of newer antiepileptics, both acute and chronic caffeine decreased the protective potential of gabapentin and topiramate but not that of lamotrigine and tiagabine. CONCLUSIONS: The existing clinical data confirm the experimental results in that caffeine intake in epileptic patients results in increased seizure frequency. It may be concluded that epileptic patients should limit their daily intake of caffeine. PMID- 18409415 TI - [Usefulness of MR CSF flow examination in children with hydrocephalus]. AB - BACKGROUND: MR examination allows the determination of the absence or presence of CSF-flow as well as its direction and velocity. THE AIM OF THE STUDY: is the presentation of the value of MR examination CSF-flow and its usefulness in diagnosis of hydrocephalus in children. MR CSF-flow examination may be performed in the cerebral aqueduct of Sylvius and in the drain of the ventriculo-peritoneal shunt. METHODS: The examination big in classical MR examination: axial and sagittal T1-weighted scans (TR 500, TE 12) and T2-weighted scans (TR 5000, TE 100). The CSF flow examination is performed in 2D CINE/GR/PC (TR 33, TE 9.1, FA 30 degrees, FOV 12 x 12 cm, slice thickness 5 mm, acquisition matrix 256 x 192) in the plane perpendicular to the examined structure. The gating sequence is compatible with the patient's heart rate. Two regions of interest is established: area of flow (ROI 1) and the area of background (ROI 2). CSF flow is calculated using Flow Analysis software. CSF-flow in CINE phase-contrast technique is used to qualitative assessment of flow. CONCLUSIONS: MR examination allows the determination of the absence or presence of CSF-flow and its direction and velocity in the cerebral aqueduct of Sylvius and in the drain of the ventriculo peritoneal shunt. This examination enables the differentiation between the shunt dependent and shunt-independent type of hydrocephalus, which is of great significance in clinical management of patients after a shunt implantation. PMID- 18409416 TI - [Physical and other methods therapy of the spasticity in children]. AB - BACKGROUND: Spasticity is a very bothering symptom, which aggravates disability and prevents in many cases a successful treatment. THE AIM: The aim of this paper was to remind different methods of treatment of spasticity, with a special attention to neurorehabilitation. RESULTS: In the research the complex pathogenesis of spasticity was referred, as well as factors influenced its aggravation and associated symptoms. Methods of evaluation of degree of spasticity were also recalled. The advantage and disadvantage of neurorehabilitation were also discussed. NDT-Bobath method as well as other kinesiotherapy methods were described with the necessity to make individualised program. The methods of rehabilitation which help in the reduction of muscle tone were refered, as well as the importance of botulin toxin, in the context of lack of efficacy of drugs in the reduction of spasticity, which cannot in fact help to reach better results of rehabilitation. The effectiveness of spasticity therapy with baclofen pump and other surgical methods were also mentioned. CONCLUSIONS: The decision to treat spasticity must be justified and depended on its intensity. It is necessary to develop new methods to obtain the durable effect of therapy of spasticity. PMID- 18409417 TI - [A pediatric neuropsychological evaluation]. AB - BACKGROUND: Pediatric neuropsychology is currently one of the most expansive fields of psychology, developing between clinical neurology and other neurosciences. THE AIM OF STUDY: This work presents the main possibilities of a pediatric neuropsychological evaluation in known clinical situations and the status of the neuropsychological profession. DESCRIPTION: The necessity of evaluating the level of cognitive functioning in children from high risk groups, children with somatic, behavioural, emotional or developmental disorders was reminded. Clinical examples are described, when a neuropsychological evaluation of the child reveals important information, becoming a valuable part of the medical diagnosing process. Diagnostic methods most commonly used for assessing the level of cognitive functioning are presented, baring in mind its division into specific cognitive domains: executive functions, learning and memory, attention, visuo-motor and verbal skills. Most popular of the international and domestic IQ tests are described, including suggestions about testing children with various degrees of intellectual disability. Among the methods are tests for assessing children in age groups from 0-18. The authors described a neuropsychological interpretation of the evaluation results, including tests, behavioural observation, interviews and other diagnostic methods. Some limitations of the neuropsychological assessment methods are also mentioned, such as questionable ecological validity. CONCLUSION: the knowledge of possibilities and the necessity of the neuropsychological evaluation in children should become much more common, especially in the medical environment. The status of the neuropsychologist in Poland need a precise legal regulation. PMID- 18409418 TI - Predator diversity and ecosystem functioning: density modifies the effect of resource partitioning. AB - The link between biodiversity and ecosystem functioning is now well established, but the challenge remains to develop a mechanistic understanding of observed effects. Predator-prey interactions provide an opportunity to examine the role of resource partitioning, thought to be a principal mediator of biodiversity function relationships. To date, interactions between multiple predators and their prey have typically been investigated in simplified agricultural systems with limited scope for resource partitioning. Thus there remains a dearth of studies examining the functional consequences of predator richness in diverse food webs. Here, we manipulated a species-rich intertidal food web, crossing predator diversity with total predator density, to simultaneously examine the independent and interactive effects of diversity and density on the efficiency of secondary resource capture. The effect of predator diversity was only detectable at high predator densities where competitive interactions between individual predators were magnified; the rate of resource capture within the species mixture more than doubled that of the best-performing single species. Direct observation of species-specific resource use in monoculture, as quantified by patterns of prey consumption, provided clear evidence that species occupied distinct functional niches, suggesting a mechanistic explanation of the observed diversity effect. PMID- 18409419 TI - Habitat saturation drives thresholds in stream subsidies. AB - Understanding how abundance regulates the effects of organisms on their ecosystems remains a critical goal of ecology, especially for understanding inter ecosystem transfers of energy and nutrients. Here we examined how territoriality and nest-digging by anadromous salmon mediate trophic subsidies to stream fishes. Salmon eggs become available for consumption primarily by the digging of salmon that superimpose their nests on previous nests. An individual-based model of spawning salmon predicted that territoriality and habitat saturation produce a nonlinear effect of salmon density on numbers of available eggs to resident predators. Field studies in Alaskan streams found that higher densities of salmon produce disproportionately more eggs in stream drift and in diets of resident fishes (Arctic grayling and rainbow trout). Bioenergetics model simulations indicated that these subsidies produce substantially enhanced growth rates of trout. These results demonstrate that small changes in salmon abundance can drive large changes in subsidies to stream food webs. Thus, the ecological consequences of population declines of keystone species, such as salmon, will be exacerbated when behavior generates nonlinear impacts. PMID- 18409420 TI - Responses of a bacterial pathogen to phosphorus limitation of its aquatic invertebrate host. AB - Host nutrition is thought to affect the establishment, persistence, and severity of pathogenic infections. Nutrient-deficient foods possibly benefit pathogens by constraining host immune function or benefit hosts by limiting parasite growth and reproduction. However, the effects of poor elemental food quality on a host's susceptibility to infection and disease have received little study. Here we show that the bacterial microparasite Pasteuria ramosa is affected by the elemental nutrition of its aquatic invertebrate host, Daphnia magna. We found that high food carbon:phosphorus (C:P) ratios significantly reduced infection rates of Pasteuria in Daphnia and led to lower within-host pathogen multiplication. In addition, greater virulent effects of bacterial infection on host reproduction were found in Daphnia-consuming P-deficient food. Poor Daphnia elemental nutrition thus reduced the growth and reproduction of its bacterial parasite, Pasteuria. The effects of poor host nutrition on the pathogen were further evidenced by Pasteuria's greater inhibition of reproduction in P-limited Daphnia. Our results provide strong evidence that elemental food quality can significantly influence the incidence and intensity of infectious disease in invertebrate hosts. PMID- 18409421 TI - Phenology: response, driver, and integrator. PMID- 18409422 TI - Linkages between phenology, pollination, photosynthesis, and reproduction in deciduous forest understory plants. AB - Light availability in the understory of deciduous forests changes drastically within the growing season due to the foliage dynamics of canopy trees. Because flowering phenology, photosynthetic characteristics, and fruiting success respond to such strong seasonality in light availability, we hypothesized that understory plants in such ecosystems should describe distinct phenological groups or syndromes where "syndrome" is defined only as a set of characteristics that co occur. To identify these phenological syndromes, we studied the flowering phenology, fruit or seed set, and photosynthetic characteristics for 18 perennial understory herbaceous species that differed in reproductive strategy over eight years in a deciduous forest in northern Japan. Three phenological groups emerged from this study: (1) spring bloomers, flowering and fruiting before the completion of canopy closure; (2) early-summer bloomers, flowering during the progress of canopy closure and fruiting after canopy closure; and (3) late-summer bloomers, flowering and fruiting after canopy closure. The spring bloomers had high photosynthetic rates and high fruiting abilities, but the flowering time varied considerably among years due to yearly fluctuations of snowmelt date. Bumble bee-pollinated species of spring bloomers showed variable seed-set success, while fly-pollinated species showed relatively stable seed sets over the years. The early-summer bloomers showed low fruiting abilities irrespective of pollination success, reflecting severe resource limitation with decelerating light availability during fruit development. Although the late-summer bloomers showed low photosynthetic rates under low-light conditions, high fruit-set success was attained if pollination was sufficient. These results support our hypothesis that phenological syndromes may be found in deciduous forest understory plants. Given that reproductive success of bee-pollinated spring bloomers is highly susceptible to seasonal fluctuation, climate change may have its strongest impacts on this group. PMID- 18409423 TI - Global warming and flowering times in Thoreau's Concord: a community perspective. AB - As a result of climate change, many plants are now flowering measurably earlier than they did in the past. However, some species' flowering times have changed much more than others. Data at the community level can clarify the variation in flowering responses to climate change. In order to determine how North American species' flowering times respond to climate, we analyzed a series of previously unstudied records of the dates of first flowering for over 500 plant taxa in Concord, Massachusetts, USA. These records began with six years of observations by the famous naturalist Henry David Thoreau from 1852 to 1858, continued with 16 years of observations by the botanist Alfred Hosmer in 1878 and 1888-1902, and concluded with our own observations in 2004, 2005, and 2006. From 1852 through 2006, Concord warmed by 2.4 degrees C due to global climate change and urbanization. Using a subset of 43 common species, we determined that plants are now flowering seven days earlier on average than they did in Thoreau's times. Plant flowering times were most correlated with mean temperatures in the one or two months just before flowering and were also correlated with January temperatures. Summer-flowering species showed more interannual variation in flowering time than did spring-flowering species, but the flowering times of spring-flowering species correlated more strongly to mean monthly temperatures. In many cases, such as within the genera Betula and Solidago, closely related, co occurring species responded to climate very differently from one another. The differences in flowering responses to warming could affect relationships in plant communities as warming continues. Common St. John's wort (Hypericum perforatum) and highbush blueberry (Vaccinium corymbosum) are particularly responsive to changes in climate, are common across much of the United States, and could serve as indicators of biological responses to climate change. We discuss the need for researchers to be aware, when using data sets involving multiple observers, of how varying methodologies, sample sizes, and sampling intensities affect the results. Finally, we emphasize the importance of using historical observations, like those of Thoreau and Hosmer, as sources of long-term data and to increase public awareness of biological responses to climate change. PMID- 18409424 TI - Phenology of mixed woody-herbaceous ecosystems following extreme events: net and differential responses. AB - Ecosystem responses to key climate drivers are reflected in phenological dynamics such as the timing and degree of "green-up" that integrate responses over spatial scales from individual plants to ecosystems. This integration is clearest in ecosystems dominated by a single species or life form, such as seasonally dynamic grasslands or more temporally constant evergreen forests. Yet many ecosystems have substantial contribution of cover from both herbaceous and woody evergreen plants. Responses of mixed woody-herbaceous ecosystems to climate are of increasing concern due to their extensive nature, the potential for such systems to yield more complex responses than those dominated by a single life form, and projections that extreme climate and weather events will increase in frequency and intensity with global warming. We present responses of a mixed woody herbaceous ecosystem type to an extreme event: regional-scale pinon pine mortality following an extended drought and the subsequent herbaceous green-up following the first wet period after the drought. This example highlights how reductions in greenness of the slower, more stable evergreen woody component can rapidly be offset by increases associated with resources made available to the relatively more responsive herbaceous component. We hypothesize that such two phase phenological responses to extreme events are characteristic of many mixed woody-herbaceous ecosystems. PMID- 18409425 TI - Effects of climate change on phenology, frost damage, and floral abundance of montane wildflowers. AB - The timing of life history traits is central to lifetime fitness and nowhere is this more evident or well studied as in the phenology of flowering in governing plant reproductive success. Recent changes in the timing of environmental events attributable to climate change, such as the date of snowmelt at high altitudes, which initiates the growing season, have had important repercussions for some common perennial herbaceous wildflower species. The phenology of flowering at the Rocky Mountain Biological Laboratory (Colorado, USA) is strongly influenced by date of snowmelt, which makes this site ideal for examining phenological responses to climate change. Flower buds of Delphinium barbeyi, Erigeron speciosus, and Helianthella quinquenervis are sensitive to frost, and the earlier beginning of the growing season in recent years has exposed them to more frequent mid-June frost kills. From 1992 to 1998, on average 36.1% of Helianthella buds were frosted, but for 1999-2006 the mean is 73.9%; in only one year since 1998 have plants escaped all frost damage. For all three of these perennial species, there is a significant relationship between the date of snowmelt and the abundance of flowering that summer. Greater snowpack results in later snowmelt, later beginning of the growing season, and less frost mortality of buds. Microhabitat differences in snow accumulation, snowmelt patterns, and cold air drainage during frost events can be significant; an elevation difference of only 12 m between two plots resulted in a temperature difference of almost 2 degrees C in 2006 and a difference of 37% in frost damage to buds. The loss of flowers and therefore seeds can reduce recruitment in these plant populations, and affect pollinators, herbivores, and seed predators that previously relied on them. Other plant species in this environment are similarly susceptible to frost damage so the negative effects for recruitment and for consumers dependent on flowers and seeds could be widespread. These findings point out the paradox of increased frost damage in the face of global warming, provide important insights into the adaptive significance of phenology, and have general implications for flowering plants throughout the region and anywhere climate change is having similar impacts. PMID- 18409426 TI - Phenological sequences reveal aggregate life history response to climatic warming. AB - Climatic warming is associated with organisms breeding earlier in the season than is typical for their species. In some species, however, response to warming is more complex than a simple advance in the timing of all life history events preceding reproduction. Disparities in the extent to which different components of the reproductive phenology of organisms vary with climatic warming indicate that not all life history events are equally responsive to environmental variation. Here, we propose that our understanding of phenological response to climate change can be improved by considering entire sequences of events comprising the aggregate life histories of organisms preceding reproduction. We present results of a two-year warming experiment conducted on 33 individuals of three plant species inhabiting a low-arctic site. Analysis of phenological sequences of three key events for each species revealed how the aggregate life histories preceding reproduction responded to warming, and which individual events exerted the greatest influence on aggregate life history variation. For alpine chickweed (Cerastium alpinum), warming elicited a shortening of the duration of the emergence stage by 2.5 days on average, but the aggregate life history did not differ between warmed and ambient plots. For gray willow (Salix glauca), however, all phenological events monitored occurred earlier on warmed than on ambient plots, and warming reduced the aggregate life history of this species by 22 days on average. Similarly, in dwarf birch (Betula nana), warming advanced flower bud set, blooming, and fruit set and reduced the aggregate life history by 27 days on average. Our approach provides important insight into life history responses of many organisms to climate change and other forms of environmental variation. Such insight may be compromised by considering changes in individual phenological events in isolation. PMID- 18409427 TI - Nitrogen limitation of net primary productivity in terrestrial ecosystems is globally distributed. AB - Our meta-analysis of 126 nitrogen addition experiments evaluated nitrogen (N) limitation of net primary production (NPP) in terrestrial ecosystems. We tested the hypothesis that N limitation is widespread among biomes and influenced by geography and climate. We used the response ratio (R approximately equal ANPP(N)/ANPP(ctrl)) of aboveground plant growth in fertilized to control plots and found that most ecosystems are nitrogen limited with an average 29% growth response to nitrogen (i.e., R = 1.29). The response ratio was significant within temperate forests (R = 1.19), tropical forests (R = 1.60), temperate grasslands (R = 1.53), tropical grasslands (R = 1.26), wetlands (R = 1.16), and tundra (R = 1.35), but not deserts. Eight tropical forest studies had been conducted on very young volcanic soils in Hawaii, and this subgroup was strongly N limited (R = 2.13), which resulted in a negative correlation between forest R and latitude. The degree of N limitation in the remainder of the tropical forest studies (R = 1.20) was comparable to that of temperate forests, and when the young Hawaiian subgroup was excluded, forest R did not vary with latitude. Grassland response increased with latitude, but was independent of temperature and precipitation. These results suggest that the global N and C cycles interact strongly and that geography can mediate ecosystem response to N within certain biome types. PMID- 18409428 TI - Context matters: matrix vegetation influences native and exotic species composition on habitat islands. AB - The extensive research on plant communities of natural-habitat islands has primarily focused on the "islands." The island analogy, however, potentially limits understanding of processes influencing composition on habitat islands because the nature of their matrix is overlooked. We determine how plant community structure of the surrounding matrix influences vegetation on volcanic outcrops in the modified landscape of Banks Peninsula, New Zealand. Our primary purpose is to address whether the matrix is more important for recently established exotic species than it is for well-established native species and whether such invasion by exotics has led to homogenization of the outcrop flora. To test this, we examined our data at three spatial scales: that of the entire outcrop flora, between individual outcrops and their immediate surrounding matrix, and between individual outcrop faces and the individual releves of the immediate surrounding matrix. We found that 81% of the native flora and 90% of the exotic flora also occur in the matrix. This high level of species shared between the outcrop and matrix persists at the scale of individual outcrop faces (68% of the total flora of individual faces is shared with the matrix). We predicted that floras from different outcrops would vary in their distinctiveness from their immediate matrix. We found Bray-Curtis distance coefficient values to range from 0.26 to 0.64; these were even higher at the outcrop-face scale. Variability in outcrop distinctiveness relates primarily to the outcrop face properties of area, vegetation height, and soil depth, and matrix properties of vegetation structure and vegetation heterogeneity. The effect of the vegetation structure of the matrix is more pronounced on the exotic than on the native outcrop flora. The component of composition and structure of the matrix that was independent of outcrop properties and local environment accounts for similar levels of explainable variation in total and native composition (29-31%), but considerably more (40%) in composition of exotic species. Our results support our prediction that, as the surrounding matrix becomes more modified, invasion by exotics makes outcrop vegetation less distinct from its matrix. PMID- 18409429 TI - Constitutive and induced defenses to herbivory in above- and belowground plant tissues. AB - A recent surge in attention devoted to the ecology of soil biota has prompted interest in quantifying similarities and differences between interactions occurring in above- and belowground communities. Furthermore, linkages that interconnect the dynamics of these two spatially distinct ecosystems are increasingly documented. We use a similar approach in the context of understanding plant defenses to herbivory, including how they are allocated between leaves and roots (constitutive defenses), and potential cross-system linkages (induced defenses). To explore these issues we utilized three different empirical approaches. First, we manipulated foliar and root herbivory on tobacco (Nicotiana tabacum) and measured changes in the secondary chemistry of above- and belowground tissues. Second, we reviewed published studies that compared levels of secondary chemistry between leaves and roots to determine how plants distribute putative defense chemicals across the above- and belowground systems. Last, we used meta-analysis to quantify the impact of induced responses across plant tissue types. In the tobacco system, leaf-chewing insects strongly induced higher levels of secondary metabolites in leaves but had no impact on root chemistry. Nematode root herbivores, however, elicited changes in both leaves and roots. Virtually all secondary chemicals measured were elevated in nematode induced galls, whereas the impact of root herbivory on foliar chemistry was highly variable and depended on where chemicals were produced within the plant. Importantly, nematodes interfered with aboveground metabolites that have biosynthetic sites located in roots (e.g., nicotine) but had the opposite effect (i.e., nematodes elevated foliar expression) on chemicals produced in shoots (e.g., phenolics and terpenoids). Results from our literature review suggest that, overall, constitutive defense levels are extremely similar when comparing leaves with roots, although certain chemical classes (e.g., alkaloids, glucosinolates) are differentially allocated between above- and belowground parts. Based on a meta-analysis of induced defense studies we conclude that: (1) foliar induction generates strong responses in leaves, but much weaker responses in roots, and (2) root induction elicits responses of equal magnitude in both leaves and roots. We discuss the importance of this asymmetry and the paradox of cross-system induction in relation to optimal defense theory and interactions between above- and belowground herbivory. PMID- 18409430 TI - Testing the evolution of increased competitive ability (EICA) hypothesis in a novel framework. AB - The "evolution of increased competitive ability" (EICA) hypothesis proposes that escape from natural enemies, e.g., after transcontinental introductions, alters the selection regime because costly defenses no longer enhance fitness. Such an evolutionary loss of defenses enables resources to be directed toward growth or other traits improving performance. We tested the EICA hypothesis in a novel framework in which the natural enemy is the traveler that follows its widespread host by accidental or deliberate (biocontrol) introductions. In a greenhouse experiment we used populations of Senecio vulgaris from North America, Europe, and Australia that differ in the history of exposure to the rust fungus Puccinia lagenophorae. Contrary to what is predicted by EICA, we found no evidence for increased levels of resistance to the rust fungus in plant populations with a longer history of rust exposure (Australia). Similarly, there was no evidence for reduced fecundity in these populations, although vegetative vigor, measured as secondary branching and growth rate, was lower. The maintenance of high rust resistance in populations with no (North America) or only a short history (Europe) of rust exposure is surprising given that resistance seems to incur considerable fitness costs, as indicated by the negative association between family mean resistance and fitness in the absence of disease observed for all three continents. The comparison of population differentiation in quantitative traits with estimates of differentiation in amplified fragment length polymorphic (AFLP) markers suggests that a number of fitness-related traits are under divergent selection among the studied populations. The proposed framework to test changes in the evolutionary trajectory underlying EICA can be employed in an expanded range of systems. These may include investigations on a cosmopolitan weed or crop when an antagonist is expanding its geographic range (such as our study), studies along a chronosequence of introduction time with expected increasing accumulation of natural enemies over time, or comparisons between introduced plant populations that differ in exposure time to biocontrol organisms. PMID- 18409431 TI - Transgenerational plasticity in the sea: context-dependent maternal effects across the life history. AB - Maternal effects can have dramatic influences on the phenotype of offspring. Maternal effects can act as a conduit by which the maternal environment negatively affects offspring fitness, but they can also buffer offspring from environmental change by altering the phenotype of offspring according to local environmental conditions and as such, are a form of transgenerational plasticity. The benefits of maternal effects can be highly context dependent, increasing performance in one life-history stage but reducing it in another. While maternal effects are increasingly well understood in terrestrial systems, studies in the marine environment are typically restricted to a single, early life-history stage. Here, I examine the role of maternal effects across the life history of the bryozoan Bugula neritina. I exposed maternal colonies to a common pollution stress (copper) in the laboratory and then placed them in the field for one week to brood offspring. I then examined the resistance of offspring to copper from toxicant-exposed and toxicant-naive mothers and found that offspring from toxicant-exposed mothers were larger, more dispersive, and more resistant to copper stress than offspring from naive mothers. However, maternal exposure history had pervasive, negative effects on the post-metamorphic performance (particularly survival) of offspring: offspring from toxicant-exposed mothers had poorer performance after six weeks in the field, especially when facing high levels of intraspecific competition. Maternal experience can have complex effects on offspring phenotype, enhancing performance in one life-history stage while decreasing performance in another. The context-dependent costs and benefits associated with maternally derived pollution resistance may account for why such resistance is induced rather than continually expressed: mothers must balance the benefits of producing pollution-resistant larvae with the costs of producing poorer performing adults (in the absence of pollution). PMID- 18409432 TI - Simulated predator extinctions: predator identity affects survival and recruitment of oysters. AB - The rate of species loss is increasing at a global scale, and human-induced extinctions are biased toward predator species. We examined the effects of predator extinctions on a foundation species, the eastern oyster (Crassostrea virginica). We performed a factorial experiment manipulating the presence and abundance of three of the most common predatory crabs, the blue crab (Callinectes sapidus), stone crab (Menippe mercenaria), and mud crab (Panopeus herbstii) in estuaries in the eastern United States. We tested the effects of species richness and identity of predators on juvenile oyster survival, oyster recruitment, and organic matter content of sediment. We also manipulated the density of each of the predators and controlled for the loss of biomass of species by maintaining a constant mass of predators in one set of treatments and simultaneously using an additive design. This design allowed us to test the density dependence of our results and test for functional compensation by other species. The identity of predator species, but not richness, affected oyster populations. The loss of blue crabs, alone or in combination with either of the other species, affected the survival rate of juvenile oysters. Blue crabs and stone crabs both affected oyster recruitment and sediment organic matter negatively. Mud crabs at higher than ambient densities, however, could fulfill some of the functions of blue and stone crabs, suggesting a level of ecological redundancy. Importantly, the strong effects of blue crabs in all processes measured no longer occurred when individuals were present at higher-than-ambient densities. Their role as dominant predator is, therefore, dependent on their density within the system and the density of other species within their guild (e.g., mud crabs). Our findings support the hypothesis that the effects of species loss at higher trophic levels are determined by predator identity and are subject to complex intraguild interactions that are largely density dependent. Understanding the role of biodiversity in ecosystem functioning or addressing practical concerns, such as loss of predators owing to overharvesting, remains complicated because accurate predictions require detailed knowledge of the system and should be drawn from sound experimental evidence, not based on observations or generalized models. PMID- 18409434 TI - Shading facilitates sessile invertebrate dominance in the rocky subtidal Gulf of Maine. AB - Dramatic shifts in community composition occur between vertical and horizontal rocky surfaces in subtidal environments worldwide, yet the forces mediating this transition are poorly understood. Vertical rock walls are often covered by lush, diverse communities of sessile suspension-feeding invertebrates, while adjacent horizontal substrates are dominated by algae, or corals in the tropics. Multiple factors, including light, sedimentation, water flow, and predation have been proposed to explain this pattern, but experimental tests of these hypotheses are lacking. We manipulated light level and predation to test whether variation in these mechanisms could be responsible for the shift in composition of sessile communities between vertical and horizontal surfaces in the rocky subtidal Gulf of Maine. Shaded horizontally oriented granite plots were dominated by invertebrates (e.g., ascidians, barnacles, bryozoans) after 25 months. Unshaded plots were dominated by macroalgae, which was virtually absent in shaded plots. Exclusion of grazers with cages had no effect on percent cover of invertebrates or algae. Preferential settlement of invertebrate larvae to shaded plots, due to larval behaviors such as negative phototaxis, did not seem to play a role. Shading likely affects post-settlement mortality of invertebrates by alleviating competition for space with algae, although greater abundance of micropredators in algal-dominated communities could also be important. Communities on shaded plots lacked many taxa present on natural wall communities, likely due to greater disturbance on horizontal substrates and/or lack of sufficient time for colonization of these taxa. These results suggest that light plays a key role in controlling the structure, composition, and function of shallow subtidal communities. PMID- 18409433 TI - Controls of spatial variation in the prevalence of trematode parasites infecting a marine snail. AB - Geographic variability in abundance can be driven by multiple physical and biological factors operating at multiple scales. To understand the determinants of larval trematode prevalence within populations of the marine snail host Littorina littorea, we quantified many physical and biological variables at 28 New England intertidal sites. A hierarchical, mixed-effects model identified the abundance of gulls (the final hosts and dispersive agents of infective trematode stages) and snail size (a proxy for time of exposure) as the primary factors associated with trematode prevalence. The predominant influence of these variables coupled with routinely low infection rates (21 of the 28 populations exhibited prevalence <12%) suggest broad-scale recruitment limitation of trematodes. Although infection rates were spatially variable, formal analyses detected no regional spatial gradients in either trematode prevalence or independent environmental variables. Trematode prevalence appears to be predominantly determined by local site characteristics favoring high gull abundance. PMID- 18409435 TI - Whole-lake dissolved inorganic 13C additions reveal seasonal shifts in zooplankton diet. AB - Sustained whole-lake additions of 13C-enriched dissolved inorganic carbon (DIC), intended to increase experimentally the delta13C of DIC in the epilimnion of a small lake with high dissolved organic carbon (DOC), were made during three seasonal periods (spring, summer, and autumn). Coupled with carbon and nitrogen stable isotope analysis of zooplankton and several of their putative food sources, these additions were used to investigate seasonal changes in the relative contributions of different food sources to zooplankton diet in the lake. Four main potential food sources were considered: phytoplankton, heterotrophic bacteria (HB), methanotrophic bacteria (MOB), and green sulfur bacteria (GSB). Because the number of potential food sources exceeded the number of isotopes analyzed, a computer program (IsoSource) was used to estimate the range of possible contributions of the various food sources. During all three periods the added inorganic 13C quickly increased the epilimnetic DIC delta13C by between 18 per thousand and 21 per thousand above the initial value of approximately -21 per thousand. This 13C enrichment of DIC was rapidly transmitted to the particulate organic matter (POM), which included photosynthetic phytoplankton. In spring and summer, delta13C of both adult and juvenile Daphnia increased by approximately 10 per thousand, indicating that Daphnia utilized autochthonous carbon. However, this 13C labeling of Daphnia was not so obvious during the autumn period, when their delta13C generally decreased. According to the IsoSource model outputs based on both delta13C and delta15N values, Daphnia utilized all four potential food source types during spring, summer, and autumn, but in different proportions. The possible contribution of phytoplankton to Daphnia diet was substantial (25-71%) in all seasons. The possible contributions of the bacterial food sources were more variable. The possible contribution of GSB was minor (0 20%) at all times and negligible in autumn. The possible contribution of HB was higher but very variable. Methanotrophic bacteria always made a significant contribution to Daphnia diet and were likely the single most important food source in autumn. Since both HB and MOB in this high-DOC lake probably depend largely on allochthonous organic carbon, our results highlight the seasonal variability in the potential importance of ecosystem subsidies in lake food webs. PMID- 18409436 TI - Species size and distribution jointly and differentially determine diatom densities in U.S. streams. AB - Among the most studied relationships in ecology are those of population density with (1) body size and (2) species distribution. The first relationship, in conjunction with metabolic rate, determines the energy flows through species communities, whereas the second relationship shows how local communities are influenced by the species history of dispersal and establishment. Traditionally, these two relationships have been examined separately. Here, I explored how diatom density was affected by cell size (biovolume) and species distribution in benthic and planktonic stream habitats all the way from individual localities and hydrologic systems (regions) to the entire United States. At all scales, density was predominantly a negative function of biovolume and a positive function of distribution. Biovolume was more strongly related to density in the benthos than in the phytoplankton. Partial regressions revealed that biovolume, by itself, explained a substantially higher percentage of the variance in density at local than at regional and continental scales. Conversely, species distribution was a much more important descriptor of density at larger scales and a slightly better predictor than biovolume at local scales. At large scales density was explained primarily by distribution and, to a lesser extent and only in the benthos, by the covariance of distribution and biovolume, whereas biovolume was a marginal predictor in all habitats. This discovery suggests that the strong relationships between density and body size, reported for populations ranging from unicellular algae to mammals, may be less direct than previously thought but mediated by large-scale species distributions. PMID- 18409437 TI - Maintenance of variable responses for coping with wetland drying in freshwater turtles. AB - Aquatic animals inhabiting temporary wetlands must respond to habitat drying either by estivating or moving to other wetlands. Using radiotelemetry and capture mark recapture, we examined factors influencing the decisions made by individuals in a population of freshwater turtles (Chelodina longicollis) in response to wetland drying in southeastern Australia. Turtles exhibited both behaviors, either remaining quiescent in terrestrial habitats for variable lengths of time (terrestrial estivation) or moving to other wetlands. Both the proportion of individuals that estivated terrestrially and the time individuals spent in terrestrial habitats increased with decreasing wetland hydroperiod and increasing distance to the nearest permanent wetland, suggesting behavioral decisions are conditional or state dependent (i.e., plastic) and influenced by local and landscape factors. Variation in the strategy or tactic chosen also increased with increasing isolation from other wetlands, suggesting that individuals differentially weigh the costs and benefits of residing terrestrially vs. those of long-distance movement; movement to other wetlands was the near universal strategy chosen when only a short distance must be traveled to permanent wetlands. The quality of temporary wetlands relative to permanent wetlands at our study site varies considerably and unpredictably with annual rainfall and with it the cost-benefit ratio of each strategy or tactic. Residency in or near temporary wetlands is more successful during wet periods due to production benefits, but movement to permanent wetlands is more successful, or least costly, during dry periods due to survival and body condition benefits. This shifting balance may maintain diversity in response of turtles to the spatial and temporal pattern in wetland quality if their response is in part genetically determined. PMID- 18409438 TI - Mechanistic analysis of interspecific competition using foraging trade-offs: implications for duck assemblages. AB - The role of interspecific competition and resource partitioning in determining the composition of species assemblages is often controversial. In many cases data on species co-occurrence or resource use (prey or habitat) have been interpreted without a clear understanding of how, or even whether, phenotypic differences constrain performance to allow resource partitioning or how these constraints and the density of resources and competitors should shape resource selection by each species. Instead, predictions have been based on assumed constraints, possibly leading to conflicting results. One such controversy involves the role of bill morphology in mediating resource partitioning among dabbling ducks (Anas spp.). To determine whether incorrect assumptions may have contributed to this controversy, I constructed mechanistic models that predict filter-feeding performance for seven species of ducks directly from bill morphology and kinetics and compared these predictions to those of earlier studies that tested the bill morphology hypothesis. The models predicted that species should share a preference for their most profitable (primary) prey while partitioning their less profitable (secondary) prey by size. Consequently, ducks should forage in the same habitats and exhibit high overlap in prey size when competitor/resource ratios are either high or low. In contrast, earlier studies expected that resource partitioning should always be evident, which implicitly assumes that species partition their primary resources. The models also predicted that the ecological similarity of species in assemblages should increase as prey abundance and size variability declines, contrary to the expectations of an earlier study. A more consistent understanding of the mechanisms regulating assemblages of dabbling ducks, and other species, might emerge if patterns of resource use and species co-occurrence were predicted directly from a mechanistic understanding of how performance trade-offs affect resource selection in the context of varying resource and competitor densities. PMID- 18409439 TI - Cavity excavation and enlargement as mechanisms for indirect interactions in an avian community. AB - Direct and indirect species interactions within ecological communities may play a strong role in influencing or maintaining community structure. Complex community interactions pose a major challenge to predicting ecosystem responses to environmental change because predictive frameworks require identification of mechanisms by which community interactions arise. Cavity-nesting communities are well suited for mechanistic studies of species interactions because cavity nesters interact through the creation of and competition for cavity-nest sites. In this study, we use a cavity-nest web as a predictive framework for identifying potential indirect species interactions within a cavity-nesting community. From 2002 to 2005, we monitored abundance and nests of cavity-nesting birds in the longleaf pine (Pinus palustris) ecosystem. Using a nest-web approach, we identified a potential indirect interaction between the Red-cockaded Woodpecker (Picoides borealis) and large secondary cavity nesters, mediated by the Northern Flicker (Colaptes auratus). We used structural equation modeling to test a path model of this interaction, using cavity excavation and enlargement as mechanisms which drive the relationship between these species. Through experimental manipulation of cavity availability, we blocked links described in our model, confirming cavity creation and enlargement as processes that influence community structure. We found that a single-species management technique could potentially disrupt this indirect relationship by affecting Northern Flicker cavity excavation behavior. This study is the first demonstration of how experimental cavity manipulation can be used to test inferred processes derived from a nest web and highlights the need to understand how mechanisms underlying species interactions can complicate ecosystem responses to environmental change. PMID- 18409440 TI - Consumer resource matching in urbanizing landscapes: are synanthropic species over-matching? AB - Population responses of synanthropic species to urbanization may be explained by the resource-matching rule, which postulates that individuals should distribute themselves according to resource availability. According to the resource-matching rule, urban habitats will contain greater densities if they provide better resources than rural habitats. However, because resource availability is density dependent, individuals in urban areas would ultimately achieve fitness levels comparable to, but no better than, individuals in less urban areas. Some ecologists suggest that synanthropic birds may not conform to the resource matching rule and may instead overmatch (i.e., overexploit) in urban habitats, ultimately leading to lower fitness despite greater resource levels. Using the Northern Cardinal (Cardinalis cardinalis) as a focal species, we evaluated if Cardinal populations in urban and rural habitats were consistent with predictions of consumer resource matching. During 2003-2006 we documented population density, adult body condition, apparent survival, and annual reproductive productivity of Cardinals in riparian forest stands within urban (n = 8 stands) and rural (n = 6 stands) landscapes in Ohio, USA. Density of Cardinals in urban forests was four times that found in more rural forests. Mark-resight data from 147 males and 125 females over four years indicated that apparent survival rates were similar between urban and rural landscapes (phi = 0.64, SE = 0.039 for males and phi = 0.57, SE = 0.04 for females). Similarly, body condition indices of 168 males, 142 females, and 118 nestlings did not differ significantly between landscapes. Annual reproductive productivity (mean number of fledglings per pair over breeding season) of 294 pairs was comparable for urban (2.4 +/- 0.18 [mean +/- SE] and rural (2.1 +/- 0.18) young birds. Thus, contrary to recent suggestions, we find that high densities of certain synanthropic species in urban landscapes are consistent with expectations of consumer resource matching. PMID- 18409441 TI - An empirical evaluation of the insurance hypothesis in diversity-stability models. AB - An important stabilizing mechanism in most diversity stability models is the insurance hypothesis, which involves correlation/covariance relationships among species. These models require that species do not fluctuate synchronously over time: that is, the correlation between pairs of species does not equal 1.0. However, the strength of this stabilizing mechanism increases as correlations decline away from 1.0, especially as they become more negative and also as the summed covariance across all species pairs becomes more negative. We evaluated the importance of the insurance hypothesis as a stabilizing mechanism by examining a variety of terrestrial assemblages using long-term data from the Global Population Dynamics Database, the Breeding Bird Survey, and a long-term site in southeastern Arizona, USA. We identified co-occurring assemblages of species and calculated the Spearman rank correlations of all pairs of species and the summed covariance of the entire assemblage. We found that, in most assemblages, positive correlations were two to three times more common than negative and that the magnitude of the positive correlations tended to be stronger than the negative correlations. For all but three assemblages, the summed covariance was positive. Data from larger spatial scales tended to exhibit more positive correlations, but even at the smallest spatial scales, positive correlations outnumbered negative. We suggest that species often covary positively because coexisting species respond similarly to fluctuations in their resource base driven by climatic fluctuations. As such, our review suggests that the insurance hypothesis may not be a strong mechanism stabilizing fluctuations in natural terrestrial communities. PMID- 18409442 TI - Field-scale roles of density, temperature, nitrogen, and predation on aphid population dynamics. AB - Robust analyses of noisy, stage-structured, irregularly spaced, field-scale data incorporating multiple sources of variability and nonlinear dynamics remain very limited, hindering understanding of how small-scale studies relate to large-scale population dynamics. We used a novel, complementary Bayesian and frequentist state-space model analysis to ask how density, temperature, plant nitrogen, and predators affect cotton aphid (Aphis gossypii) population dynamics in weekly data from 18 field-years and whether estimated effects are consistent with small-scale studies. We found clear roles of density and temperature but not of plant nitrogen or predators, for which Bayesian and frequentist evidence differed. However, overall predictability of field-scale dynamics remained low. This study demonstrates stage-structured state-space model analysis incorporating bottom-up, top-down, and density-dependent effects for within-season (nearly continuous time), nonlinear population dynamics. The analysis combines Bayesian posterior evidence with maximum-likelihood estimation and frequentist hypothesis testing using average one-step-ahead residuals. PMID- 18409443 TI - Bayesian methods for analyzing movements in heterogeneous landscapes from mark recapture data. AB - Spatially referenced mark-recapture data are becoming increasingly available, but the analysis of such data has remained difficult for a variety of reasons. One of the fundamental problems is that it is difficult to disentangle inherent movement behavior from sampling artifacts. For example, in a typical study design, short distances are sampled more frequently than long distances. Here we present a modeling-based alternative that combines a diffusion-based process model with an observation model to infer the inherent movement behavior of the species from the data. The movement model is based on classifying the landscape into a number of habitat types, and assuming habitat-specific diffusion and mortality parameters, and habitat selection at edges between the habitat types. As the problem is computationally highly intensive, we provide software that implements adaptive Bayesian methods for effective sampling of the posterior distribution. We illustrate the modeling framework by analyzing individual mark-recapture data on the Glanville fritillary butterfly (Melitaea cinxia), and by comparing our results with earlier ones derived from the same data using a purely statistical approach. We use simulated data to perform an analysis of statistical power, examining how accuracy in parameter estimates depends on the amount of data and on the study design. Obtaining precise estimates for movement rates and habitat preferences turns out to be especially challenging, as these parameters can be highly correlated in the posterior density. We show that the parameter estimates can be considerably improved by alternative study designs, such as releasing some of the individuals into the unsuitable matrix, or spending part of the recapture effort in the matrix. PMID- 18409444 TI - The factorial decomposition of the Mahalanobis distances in habitat selection studies. AB - The Mahalanobis distances have been introduced in habitat selection studies for the estimation of environmental suitability maps (ESMs). The pixels of raster maps of a given area correspond to points in the multidimensional space defined by the mapped environmental variables (ecological space). The Mahalanobis distances measure the distances in this space between these points and the mean of the ecological niche (i.e., the hypothesized optimum for the species) regarding the structure of the niche. The map of these distances over the area of interest is an estimated ESM. Several authors recently noted that the use of a single optimum for the niche of a species may lead to biased predictions of animal occurrence. They proposed to use instead a minimum set of basic habitat requirements, found by partitioning the Mahalanobis distances into a restricted set of biologically meaningful axes. However, the statistical approach they proposed does not take into account the environmental conditions on the area where the niche was sampled (i.e., the environmental availability), and we show that including this availability is necessary. We used their approach as a basis to develop a new exploratory tool, the Mahalanobis distance factor analysis (MADIFA), which performs an additive partitioning of the Mahalanobis distances taking into account this availability. The basic habitat requirements of a species can be derived from the axes of the MADIFA. This method can also be used to compute ESMs using only this small number of basic requirements, therefore including only the biologically relevant information. We also prove that the MADIFA is complementary to the commonly used ecological-niche factor analysis (ENFA). We used the MADIFA method to analyze the niche of the chamois Rupicapra rupicapra in a mountainous area. This method adds to the existing set of tools for the description of the niche. PMID- 18409445 TI - The top-down mechanism for body-mass-abundance scaling. AB - Scaling relationships between mean body masses and abundances of species in multitrophic communities continue to be a subject of intense research and debate. The top-down mechanism explored in this paper explains the frequently observed inverse linear relationship between body mass and abundance (i.e., constant biomass) in terms of a balancing of resource biomasses by behaviorally and evolutionarily adapting foragers, and the evolutionary response of resources to this foraging pressure. The mechanism is tested using an allometric, multitrophic community model with a complex food web structure. It is a statistical model describing the evolutionary and population dynamics of tens to hundreds of species in a uniform way. Particularities of the model are the detailed representation of the evolution and interaction of trophic traits to reproduce topological food web patterns, prey switching behavior modeled after experimental observations, and the evolutionary adaptation of attack rates. Model structure and design are discussed. For model states comparable to natural communities, we find that (1) the body-mass abundance scaling does not depend on the allometric scaling exponent of physiological rates in the form expected from the energetic equivalence rule or other bottom-up theories; (2) the scaling exponent of abundance as a function of body mass is approximately -1, independent of the allometric exponent for physiological rates assumed; (3) removal of top-down control destroys this pattern, and energetic equivalence is recovered. We conclude that the top-down mechanism is active in the model, and that it is a viable alternative to bottom-up mechanisms for controlling body-mass-abundance relations in natural communities. PMID- 18409446 TI - Biomass and toxicity responses of poison ivy (Toxicodendron radicans) to elevated atmospheric CO2: comment. PMID- 18409447 TI - Top 10 things every healthcare security director must get right! AB - For the security professional there is nothing unfamiliar about the ten issues presented by the author. Getting them right, he claims, is another matter. The article provides many recommendations and suggestions for performance improvement and liability avoidance. But the greatest opportunities for achieving success will be found in the answers that the reader gives to the host of significant questions posed. PMID- 18409448 TI - Gang culture from the streets to the emergency department. AB - Gangs, gang activity, and the results of such activities, can have a special impact on hospitals, their employees, and others in the hospital community. In this article, the authors spell out in detail the extent of the gang culture to assist hospital professionals in understanding its many ramifications and guide them in taking actions to reduce the impact of gangs on hospital operations. PMID- 18409449 TI - A visitor control system for patients who were victims of violent crimes. AB - The dangers of further violence to patient victims of gang-related crime as well as violence against members of their families are frankly discussed in this article by the author who describes a special visitor control system he developed to prevent such violence from spreading into the hospital. PMID- 18409450 TI - Police encounters of the worst kind. AB - Incidents where prisoners are taken to a local hospital for treatment and use the opportunity to try to escape are rare, but not rare enough, the author reports, citing incidents where police, security officers, hospital personnel, or citizens were killed, wounded, or otherwise injured when the patient prisoner resorted to violence. The article presents a program in which police and hospital authorities can cooperate in doing a better job of preventing such incidents. PMID- 18409452 TI - Verbal judo training of a security staff: a comprehensive cancer center's experience. AB - How a large hospital with over 250 security officers improved the communications practices of its line officers through a training program that successfully adapted the Verbal Judo Course to the specific needs and requirements of hospital security in-service training. PMID- 18409451 TI - Developing an emergency code response team for behavioral issues. AB - An emergency code response team is seen as the answer by the author when intervention issues arise in connection with violence or potential violence by patients or others in the hospital. In the article he explains what is involved in recruiting and training such a team, the roles of its members in a crisis situation, and the preparation and follow-up necessary to perform its duties successfully. PMID- 18409453 TI - Who's in your facility? Screening and managing vendors. AB - This two perspective article addresses the issue of vendor screening and identification. It describes a process now in place at a hospital that complies with new Federal regulations for screening and managing vendors who enter a healthcare facility. PMID- 18409454 TI - Serial murder by healthcare professionals. AB - The prosecution of Charles Cullen, a nurse who killed at least 40 patients over a 16-year period, highlights the need to better understand the phenomenon of serial murder by healthcare professionals. The authors conducted a LexisNexis search which yielded 90 criminal prosecutions of healthcare providers that met inclusion criteria for serial murder of patients. In addition we reviewed epidemiologic studies, toxicology evidence, and court transcripts, to provide data on healthcare professionals who have been prosecuted between 1970 and 2006. Fifty four of the 90 have been convicted; 45 for serial murder, four for attempted murder, and five pled guilty to lesser charges. Twenty-four more have been indicted and are either awaiting trial or the outcome has not been published. The other 12 prosecutions had a variety of legal outcomes. Injection was the main method used by healthcare killers followed by suffocation, poisoning, and tampering with equipment. Prosecutions were reported from 20 countries with 40% taking place in the United States. Nursing personnel comprised 86% of the healthcare providers prosecuted; physicians 12%, and 2% were allied health professionals. The number of patient deaths that resulted in a murder conviction is 317 and the number of suspicious patient deaths attributed to the 54 convicted caregivers is 2113. These numbers are disturbing and demand that systemic changes in tracking adverse patient incidents associated with presence of a specific healthcare provider be implemented. Hiring practices must shift away from preventing wrongful discharge or denial of employment lawsuits to protecting patients from employees who kill. PMID- 18409455 TI - Security challenges and risk management strategies for child abduction. AB - Infant and child abduction is a serious risk exposure for hospitals. Such a horrific event can impose monumental injury on family members, the facility and its staff, as well as the community. Having the right security plan and liability coverage in place is essential to managing a risk, that, while not statistically common, can become high profile. PMID- 18409456 TI - Infant abduction: taking a new look at "false" alarms. AB - Complacent responses by staff to repeated "false" alarms have resulted in circumventing infant protection systems by kidnappers, reports the author, who makes the case for taking a new approach to and new attitude about the purpose and functioning of hospital alarm systems. Rather than looking at a so-called false alarm as an unnecessary nuisance, we need to see it as an opportunity to test our total alarm response system and be grateful that it worked as it was intended, he demonstrates in this article. PMID- 18409457 TI - The art and science of proactive security programming. AB - The renewed effort on the part of law enforcement and private security to prevent crimes from happening, given new emphasis after the events of 9/11, is described by the author who provides guidance in fashioning a proactive crime control program. The article also presents a number of ways for mitigating crime risk exposure. PMID- 18409458 TI - The future of security training. AB - This article reports on the brave new world of private security training, whether in healthcare or any other industry. It gives details and advice on new requirements, new approaches, and new resources for the forward-looking security director. PMID- 18409459 TI - Crisis communications. AB - Whether healthcare security is directly or indirectly involved in a crisis that will involve the media, it will pay to better familiarize yourself with the kind of planned responses discussed in this article to mitigate negative outcomes. PMID- 18409460 TI - Securing our hospitals--an executive summary of the GE security & IAHSS healthcare benchmarking study. AB - The landmark study, published in October 2007, enables hospital security directors to compare their facilities to others and use the benchmarking information to help improve their programs. In this article, the author who directed the study's research, describes what she sees as some of its key findings. The complete presentation, which covers 48 detailed charts, can be downloaded at the IAHSS website (www.iahss.org). Go to the member welcome page to access the GE Study. PMID- 18409461 TI - HEICS to HICS, some observations and experiences. AB - There has been a marked improvement in the capability of hospitals to respond to emergency situations with the introduction of HICS in 2005, the author says, citing a wealth of available materials for developing a robust response plan. He warns security directors, however, about current practices which may be keeping them out of the emergency management loop. PMID- 18409462 TI - Negligent hiring liability--a primer for healthcare administrators. AB - Healthcare institutions are particularly vulnerable to negligent hiring liability which holds employers responsible for their employees' improper conduct. Several methods are simple and inexpensive yet many employers neglect to employ them effectively, the author, an attorney and security management consultant, points out. In this article, he spells out what can be done to screen applicants to avoid and/or better defend against negligent hiring lawsuits. PMID- 18409463 TI - [Use of academic urological terminology and professional risks: a warning]. PMID- 18409464 TI - [The urodynamic "in vitro"]. PMID- 18409465 TI - [Development of nephrectomy during the nineteenth century and its introduction into Spain]. AB - INTRODUCTION AND OBJECTIVES: After first nephrectomy lumbar made by Dr. Simon in 1869, the surgical technique was established and the procedure access was followed by several surgeons with initial controversy between the use of the intra or extraperitoneal access. We carried out a review about the evolution of this surgical procedure among XIX century and of those who were pioneers with this Kind of technique in our country. MATERIAL AND METHODS: We have reviewed books of history of the Urology, the chapters of surgery in classic urology volumes and the oral presentations to the Spanish Medical congresses, until beginning of XX century and the published nephrectomy procedures in medical literature of that time. RESULTS: We analyzed different statistical studies appeared at the time about results of the operation, the decrease of mortality in the nephrectomies made by extraperitoneal access demonstrated its advantage soon. We found out 35 kidney removal performing the lumbar access, carried out in Spain in the last third of the XX century, three by Dr Federico Rubio and Gall in Madrid, in Barcelona by the urologist Dr. Victor Azcarreta Colan and Jose Pages and by the surgeons Drs. Esquerdo and Salvador Cardinal, from Sevilla, Dr Antonio Solano and the canary urologist Dr. Jose Hernandez Perez, who were pioneers performing this procedure in our country. CONCLUSIONS: The introduction of nephrectomy by lumbar access in Spain was delayed in relation to the rest of Europe, due to the secular delay that we dragged, only when adequate conditions of the operating rooms, as much as anaesthesia and asepsis and surgical devices tools, guaranteed the success operation, the main surgeon begun to perform it. PMID- 18409466 TI - [Algorithm for the treatment of patients with obstructive uropathy secondary to prostate cancer]. AB - BACKGROUND: One of the prostate cancer progression complications is the obstructive uropathy, by infiltration and compression of the distal ureteral section, that can entail to an acute renal insufficiency, with affectation of the quality of life and the survival of these patients. The treatment of prostate cancer with secondary ureterohidronefrosis is palliative and following the present tendencies, the positioning of a nephrostomy is considered. MATERIALS AND METHODS: A search was made in PUBMED and the most representative articles were reviewed. The algorithm was constructed with the daily routine clinical base, the protocol of our center and with the scientific evidence available in medical literature. RESULTS: An algorithm of decisions sets out to define the urinary derivation in patients with obstructive uropathy secondary to prostate cancer. CONCLUSIONS: The indication to place a nephrostomy in patients with obstructive uropathy secondary to prostate cancer must be approached individually, according to the general conditions and the quality of life of the patient with base in scales defined in literature (ECOG and Karnofsky) and in factors of good or bad prognosis, always considering ethical considerations and the consent of the patient and his family. PMID- 18409467 TI - [Recent changes in prostate cancer mortality in Spain. Trends analysis from 1991 to 2005]. AB - OBJECTIVE: [corrected] To describe the evolution of prostate cancer mortality in Spain during the period 1991-2005. SUBJECTS AND METHODS: A trend analysis for age standardized mortality rates (European standard population) was performed, using joinpoint regression analysis, which allows estimation of the annual percent change of rates and to find significant changes in such trend. RESULTS: Age standardized mortality rates in Spain reached their peak value in 1996. The joinpoint analysis identified two different periods in the trend of the age standardized rates: a first one of increase in rates between 1991 and 1996 (2.1% annual increase) and a second period starting in 1996, in which rates decline at an annual rate of 2.7%. CONCLUSION: The tendency of prostate cancer seen in Spain resembles that of industrialized areas, with an increase in its age standardized death rates that suffers a downturn by the end of the past decade. Mortality declines for prostate cancer are now evident in 14 out of the 17 Autonomous Communities. Increases in PSA screening and better treatment of early-stage disease, possibly acting in combination, remain plausible hypotheses. PMID- 18409468 TI - [Importance of microlithiasis in testicular germ cell tumor ultrasound]. AB - INTRODUCTION: Testicular microlithiasis (TM) is an infrequent finding in testicular ultrasound and its clinical importance has not been completely defined. We analyzed the ultrasounds of patients with testicular germ cell tumors in order to analyze the correlation between TM, histological findings and clinical variables. METHODS AND MATERIALS: Fifty-seven patients with germ cell tumors and radical orchiectomy were included. Clinical, pathological, and echographic data were analyzed. RESULTS: TM was observed in 27 men (48.27%) and was absent in 30 (52.6%). Patients with TM had a greater likelihood of nonseminomatous germ cell tumors (NSGCT) vs seminomatous (55.6% vs 30%, p=0.05), stage II/III testicular cancer (51.8% vs 16.7%, p=0.005), positive surgical margins (18.5% vs 0%, p=0.021), and spermatic cord invasion (14.8% vs 0%, p=0.048). No significant difference was found in respect to other histopathological variables. CONCLUSION: This study showed that TM in testicular tumors is associated to NSGCT, advanced clinical stage, positive surgical margins, and spermatic cord invasion. PMID- 18409469 TI - [Ancillary studies in the differential diagnosis of epithelial renal cell tumors with granular cells]. AB - INTRODUCTION AND OBJECTIVES: Differential diagnosis of renal neoplasms with granular cells may pose difficulties and implications on ontogeny and prognosis. Our aims are to characterize the pattern of immunostaining and to search for a useful diagnosis panel. METHODS: We studied with colloidal iron staining (Mowry's modified method) and 22 commonly used immunomarkers 22 conventional carcinomas (CC), 37 chromophobe carcinomas (CPC), 8 oncocytomas (OC), and 7 collecting duct carcinomas (CDC) with granular cells. Cases with not entirely clear diagnosis were excluded. RESULTS: Colloidal iron staining was diffuse, strong, reticular, and cytoplasmic in 32 CPC cases, the diffuse and strong pattern was not observed in other tumors. The more useful diagnostic panel was cytokeratin 7 (CK7)/CD10/vimentine (vim). The 8 OC were negative for the three antibodies. The most common profile for CC was CK7-/CD10+/vim+, the CPC profile was CK7+/CD10 /vim-, and CDC did not show a particular profile. With these three antibodies specificity was >90% for the differential diagnosis. Adding colloidal iron staining specificity was 100%. CONCLUSIONS: Morphologic features, colloidal iron staining (modified Mowry's method), and immunostaining with CK7/CD10/vim permit the final diagnosis with high specificity. However, a 100% specific marker does not exist at the present time. PMID- 18409470 TI - [Quality of life in patients with urinary incontinence]. AB - OBJECTIVE: To assess the quality of life of female patients with urinary stress incontinence (USI) who were attended at the urology department of the University Hospital at Albacete. METHOD: Between November 2001 and December 2005, 126 patients with USI were surveyed in our hospital using the King's Health Questionnaire (KHQ), which is a specific instrument to measure the quality of life among female patients with urinary incontinence (UI). RESULTS: The mean age of patients was 57.09 anos (SD: 9.57) and the mean BMI was 28.14 Kg/m2 (SD: 4.66). The mean evolution of UI was 114.48 months, with a median of 96 months. During the last week of the study period, a total of 73 women (57.94%) had more than 10 urine leaks a day. As regards the number of sanitary towels used a day, 82.5% (104 cases) stated they used less than 6. The urodynamic diagnosis was described as normal in 8 cases (6.3%), 16 cases (12.7%) were diagnosed as mixed urinary incontinence and, finally, 102 cases (81%) suffered genuine USI. The best scores were presented in the Personal Relationships scale with a mean score of 26.8, whereas the worse scores were noted in that of Impact of Urinary Incontinence, whose mean score was 82.96. Generally, the lowest scores (better quality of life) were obtained in the youngest age groups. Scores in the scales of General Health, Impact on daily activities, Impact on physical activity, Impact on social activity and Emotions, were significantly higher in patients who had undergone a histerectomy. Patients whose evolution of urinary incontinence was longer presented lower scores in the Personal Relationships scale (r=0.179; p=0.045). CONCLUSIONS: The quality of life of female patients with U.I. is affected, which limits both their physical activity and image. Their quality of life worsens with age, a greater extent of incontinence, greater urinary symptomatology and when episodes of urinary infection are associated. PMID- 18409471 TI - [Questionnaire for evaluation of sexual function in women with genital prolapse and/or incontinence. Validation of the Spanish version of "Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12)"]. AB - This article summarizes the work done to adapt and to validate the short form of Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) for its use in Spain. It will become the first validated questionnaire in this country for the evaluation of the sexual function in women with Pelvic Organ Prolapse and/or Urinary Incontinence. PATIENTS AND METHOD: 49 women who visited a specialized unit with symptoms of pelvic floor were included. Patients filled in the Spanish version of the questionnaire to validate (PISQ-12), the Urinary Incontinence Questionnaire (ICIQ-UI-SF); the Female Sexual Function Questionnaire (FSM) and the Bladder Control Autoevaluation Questionnaire (CACV). Factibility, reliability and validity of the new questionnaire were evaluated. RESULTS: Factibility: 99.83% of the sample answered all the items (only one patient did not answer one of the items); average administration time 3.5 (1.5) minutes. RELIABILITY: Cronbach's alpha was 0,829. VALIDITY: PISQ-12 correlation with FSM was 0,71; with ICIQ-UI-SF it was -0,038; with the CACV "symptoms" dimension the correlation was -0,30 and with the "discomfort" dimension it was -0,40. The existence of the same three dimensions of the PISQ-12 original version in the adapted Spanish questionnaire is checked through a factorial analysis. The score in PISQ-12 was worse (lower) in the case of women with Hyperactive Bladder symptoms and discomfort measured with the CACV questionnaire and in women with sexual dysfunction measured with FSM. PISQ-12 is an instrument with the appropriate psychometric characteristics to evaluate sexual function in women with pelvic floor problems. PMID- 18409472 TI - [Renal graft outcome in patients with associated liver transplant]. AB - INTRODUCTION: Nearly 50% of liver transplant recipients have some degree of renal failure; patients in haemodialysis treatment have a higher risk of suffering hepatic diseases related to viral infections or concomitant pathologies. Improvement in surgical and organ preservation techniques and immunosuppressive therapy has permitted multiorganic transplants in patients needing both liver and kidney organs. OBJECTIVES: To review our results in renal transplants in those patients with liver and kidney transplants. MATERIAL AND METHOD: Retrospective study of the 15 patients with liver and kidney transplants performed in our Hospital. We have reviewed patients main characteristics, liver and renal failure causes, renal graft and patient outcome and complications relate to renal transplant. RESULTS: Between 1975 and December 2006 we performed 1483 kidney transplants and between 1991 and December 2006, 409 liver transplants. We performed multiorganic liver and kidney transplants to 15 patients (4 women and 11 men). The average for liver transplant recipients was 52.5+/-9.3 years (range 37-61) and for kidney transplant recipients was 51+/-12.5 years (35-66). Cold ischemia was 6.4+/-5.4 hours (6-8) in simultaneous liver-kidney transplant and 20.5+/-5.4 (8-27 hours) in non-simultaneous ones. Three patients had a renal transplant before the liver one (two functioning which had no changes after hepatic transplant but the other was lost due to IgA glomeruloneprhitis relapse and received a simultaneous kidney-liver transplant). Six patients received a simultaneous kidney-liver transplant and eight patients a renal transplant between 16 and 83 months (x=50.5+/-25.9 months) after the liver transplant. A renal graft was lost due to renal vein thrombosis and two due to IgA relapse; the others were functioning between 6 and 264 months of follow-up (x=92.5+/-66.7) with creatinine levels of 1.86+/-mg/100, (range 1-4.5). Four patients died due to hepatic failure between 8 months and 21 years after renal transplant and another died of oesophagus cancer 14 years after the kidney transplant, in all cases with functioning renal graft. There were no cases of kidney graft acute rejection in simultaneous transplants but there were five in non-simultaneous ones. Immunotherapy was based on steroids and tacrolimus. CONCLUSIONS: Liver-kidney transplants are worthy options in patients with hepatic and renal end failure. Acute rejection seems to have fewer incidences in simultaneous liver-kidney transplantation. PMID- 18409473 TI - [The use of double J stent for prevention of urological complications in kidney transplants]. AB - OBJECTIVE: To analyze the impact of the use of the double J stent in the incidence of urological complications, like fistula urinary and ureteral obstruction in kidney transplants. MATERIALS AND METHODS: Between August 2003 and December 2005, 59 adult recipients underwent renal transplant. A retrospective study was conducted on two groups of patients: Group A with double J stent and group B without it. We reviewed the urological complications: fistula, ureteral obstruction and urinary tract infection. RESULTS: Group A: One ureteral obstruction and two urinary fistulas (7%) were developed in this group. 13 patients (46%) had a positive urinary culture. In only one case was necessary to retire the double J stent because of ureteral obstruction. Group B: Four patients (13%) developed ureteral obstruction and another five (16%) developed urinary fistula. 9 patients (29%) had a positive urinary culture. CONCLUSION: The routine insertion of a double J stent in kidney transplants reduces the number of early complications urinary fistula and ureteral obstructions. PMID- 18409474 TI - [Validation of the Spanish version of the Benign Prostatic Hyperplasia Impact Index Questionnaire. "Validart Study"]. AB - INTRODUCTION: Benign Prostatic Hyperplasia is a chronic disease that affects Health related quality of life (HRQL). In Spain there are no specific questionnaires to assess HRQL in patients suffering from BPH. The objective of this study was to validate the Spanish version of the Benign Prostatic Hyperplasia Impact Index (BII) questionnaire. MATERIAL AND METHODS: Study participants were men over 49 with confirmed BPH diagnosis and evaluated in three visits. Psychometric properties of the questionnaire were analyzed in terms of validity, reliability and sensitivity to change. RESULTS: A total of 1,719 men with mean (SD) age 68.36 were evaluated and classified according to the BPH severity. Both longitudinal and construct validity (r>0.6) of the BII questionnaire were moderate-high. Internal consistency (alpha=0.91), test-retest reliability (CCI=0.8) and sensitivity to change (effect size=0.6) showed good results. The HRQL improved significantly during the monitoring period (p<0.01). DISCUSSION: There were no available specific HRQL questionnaires for Spanish population with BPH until now. The Spanish version of the BII questionnaire fulfils with the psychometric properties that are necessary to evaluate HRQL in patients with BPH; furthermore, it has been confirm the considerable impact of this disease in the quality of life of this patients. PMID- 18409476 TI - [Obstructive symptoms in young woman by left hydrosalpinx]. AB - We describe the case of a 43 years old woman with obstructive and irritative symptoms after having an hysterectomy. Bladder outlet obstruction was observed in an urodinamyc study. She was later diagnosed with left hidrosalpinx. Salpingectomy was performed, as a result all symptoms have disap- PMID- 18409477 TI - [Horseshoes kidney isthmus carcinoma. A case report]. AB - Horseshoe kidney is the most frequent fusion abnormality of the kidney. The incidence of renal carcinoma in patients with horseshoe kidney is similar to those with normal anatomy. Its special anatomical features must be borne in mind for both surgical approach and conservative surgery. We present a horseshoe kidney isthmus carcinoma case report in which we performed conservative surgery of both renal units. PMID- 18409478 TI - [Bladder infection by schistosoma: an uncommon cause of haematuria]. AB - Immigration is a real phenomenon in our country with direct consequences into the public health system. In the attendance to these patients we do not have to forget the existence unusual diseases in our setting. We presented the clinical case of a patient with hematuria caused by bladder schistosomiasis. PMID- 18409479 TI - [Ectopic ureter as cause of pyonephrosis and urinary incontinence]. AB - Ectopic ureter accounts with an incidence of 1 in 2000 newborns. When present, ectopic ureter can be associated with duplex kidneys in an 85 % of the cases. Clinical manifestations of this malformation include incontinence and urinary tract infections. Ectopic ureter frequently occurs in association with a dysplastic upper pole renal moiety. When a poorly functioning upper pole segment is present, a standard surgical treatment is upper pole heminephrectomy. A 23 years old woman presented with left renal colic pain, fever and urinary leak. Ultrasound, intravenous pyelogram and antegrade pyelogram revealed a partial duplex right kidney and a complete duplex left kidney with hydronephrosis and ectopic insertion into the urethra of the left upper pole moiety. Following diagnosis upper pole heminephrectomy and partial ureterectomy was performed. PMID- 18409480 TI - [Scrotal mass as result of the extension of a pancreatic pseudocyst]. AB - There are several causes for orchitis and among them there are inflammatory process. We want to communicate the case of a young man with a clinical history and physical examination compatible with orchitis produced by the extension of a pancreatic pseudocyst that was not suspected in the beginning and discovered by imaging procedures. PMID- 18409481 TI - [Biourology.com]. PMID- 18409482 TI - [Obtape sling complication]. PMID- 18409483 TI - [Obstructive fungus ball in kidney allografts]. PMID- 18409484 TI - [Two patients with testicular microlithiasis]. PMID- 18409485 TI - [Large ureteral stone]. PMID- 18409486 TI - [The results of dynamic observation over patients with stomach and duodenum ulcer diseases after Helicobacter pylori eradication]. PMID- 18409487 TI - [Metabolism of acetylcholine and clinical characteristics of stomach ulcer]. PMID- 18409488 TI - [Argon-plasmic coagulation in the local treatment of acute erosive-ulcer damages of the stomach and duodenum in conditions of multiple-discipline hospital]. PMID- 18409489 TI - [Grounds and perspectives of using geviskon--novel in Russia alginate-containing drug--in treatment of acid-dependent diseases of the digestive system]. PMID- 18409490 TI - [ACE inhibitors in the treatment of patients with arterial hypertension and pathology of the digestive system]. PMID- 18409491 TI - [Morphologic and functional changes of upper parts of the digestive tract in patients with chronic diffuse diseases of the liver with portal hypertension according to endoscopy, pH-data, and bacteriological tests for helicobacteriosis]. PMID- 18409492 TI - [Difficulties of differential diagnostics of inflammatory intestinal diseases]. PMID- 18409493 TI - [Correction of functional and structural changes of the intestine by ursosan in patients with absence of the gall-bladder]. PMID- 18409494 TI - [New approach to diagnostics and correction of portohepatic hemodynamics in patients with chronic liver diseases]. PMID- 18409495 TI - [State of the muscular layer of the stomach fundal section during feeding with dispersed food (according to the experimental and morphological study)]. PMID- 18409496 TI - [Role of functional state of the respiratory system and risk factors in development of pneumonia after surgical procedures on the upper part of the abdominal viscera]. PMID- 18409497 TI - [Problems of diagnostics and treatment of isolated and associated liver trauma]. PMID- 18409498 TI - [Morphological changes of the pyloric sphincter in pylorostenosis of patients with ulcer disease]. PMID- 18409499 TI - [H1-receptors: spontaneous activity, agonism, and reversed agonism]. PMID- 18409500 TI - [Using pollen in the complex treatment of patients with malnutrition]. PMID- 18409501 TI - [Complex approach to the differential diagnostics of chronic pancreatitis: results of radio-visualization, mini-invasive interventions with ultrasonic control, and quality of life assessment]. PMID- 18409502 TI - [Antiviral therapy of patients with chronic hepatitis C with normal level of aminotransferases]. PMID- 18409503 TI - [Endometriosis of the sigmoid, complicated with relapsing intestinal obstruction in the large intestine]. PMID- 18409504 TI - [Imaging of gout]. PMID- 18409505 TI - [Gout and hyperuricemia: Introduction]. PMID- 18409506 TI - [Historical review of gout and hyperuricemia investigations]. AB - Historical development of gout and hyperuricemia investigations was reviewed. Gout has been a recognized disease since the fifth century B.C. In 1683, Sydenham described the detailed clinical features of the disease based on his own condition. Leeuwenhoek (1679) first described crystals in a gouty tophus, which were identified as uric acid by Wollaston (1797). Since uric acid clearance of hyperuricemia was markedly lower than that in normal controls, early investigators considered that the main cause of hyperuricemia was urate underexcretion. However, in the 1940s, studies on uric acid metabolism using isotope tracer techniques demonstrated that a part of hyperuricemia resulted from urate overproduction, which was detected in approximately one-third of all gouty patients. In the 1970s, micropuncture, microinjection and microperfusion methods as well as stop-flow methods demonstrated that uric acid transports in nephron were suspected to consist of four steps, that were glomerular filtration, reabsorption, secretion and postsecretory reabsorption. The majority of filtrated uric acid was almost completely reabsorbed, followed by secretion and postsecretory reabsorption at a proximal site in the tubulus. Each proportion of transports to the glomerular filtration(100%) was estimated approximately 99%, 50% and 40%, respectively. Subsequently, about 10% of the filtrate was excreted in the urine. The authors (1999) suggested that the secretion rate of hyperuricemic patients was significantly lower than that of normal controls but postsecretory reabsorption was not. Therefore, the decrease in the secretion rate was suspected to be the main cause of underexcretion. Dunkan (1960) reported a family demonstrating hyperuricemia associated with severe renal damage that progressed rapidely. Currently, this disease is called familial juvenile hyperuricemic nephropathy (FJHN), and was recently found to be the result of a variation in uromodulin. Enomoto (2002) found a number of urate transporters in the cell surface of the tubulus, among which URAT1 was the most effective in reabsorbing urate from the tubulus lumen to the cells. The urate was released to the blood vessel side by the other transporter OAT. Therefore, URAT1 was suspected to be a cause of underexcretion. As the mechanism underlying overproduction of uric acid, de novo purine nucleotide synthesis has been shown to be increased. In some cases, the increase in de novo synthesis is the result of gene mutation in purine nucleotide synthesis enzymes, such as PRPP synthetase (Sperling, 1973) as well as hypoxanthine guanine phosphoribosylpyrophosphate synthetase (Seegmiller, 1967). However, the mechanism in majority of the overproduction has not yet been clarified and is currently under investigation. PMID- 18409507 TI - [Definition and classification of hyperuricemia]. AB - Hyperuricemia (serum urate level >7 mg/dL) is caused by accelerated generation of uric acid and/or impaired excretion in the kidney. It is classified into three types (overproduction, underexcretion and mixed types). The typical cases with the type of uric acid overproduction are hypoxanthine guanine phosphoribosyl transferase deficiency due to HPRT gene abnormality, excessive consumption of purine-rich diet, and cytolysis induced by chemotherapy for blood neoplasm, those with the type of underexcretion are familial juvenile hyperuricemic nephropathy due to uromodulin gene abnormality and abrupt body weight loss due to low calorie diet, and those with the mixed type are glucose 6-phosphatase deficiency due to glucose 6-phosphatase gene abnormality and excessive consumption of alcohol beverages. PMID- 18409508 TI - [Revised version of Guideline for the Management of Hyperuricemia and Gout]. AB - Guideline for the management of hyperuricemia and gout, first edition was published in 2002 from Japanese Society of Gout and Nucleic Acid Metabolism, and has contributed for the improvement in the management of patients by primary physicians. During five years after the publication of this first edition, large amount of evidences has been accumulated, thus it is time to revise the guideline. Japanese Society of Gout and Nucleic Acid Metabolism has organized the committee for the revision of guideline, and the operations have begun. In this revision, we pay attention to the quality in the process of guideline edition, application of AGREE checklist and patients perspective is considered. PMID- 18409509 TI - [Epidemiology of hyperuricemia and gout in Japan]. AB - It has been reported that the prevalence of hyperuricemia in Japan is 20-25% in adult male population and has been increasing at least until a few years ago. Although epidemiological data for the prevalence of gout in Japan is limited, a recent study conducted in a local area of Wakayama prefecture reported a prevalence of 1.1% in men. Japanese national census data revealed that the number of individuals visiting hospitals with a self-diagnosis of gout has been increasing. The census data also showed that the proportion of individuals with obesity(BMI > or = 25) has been increasing in male population, probably due to decreasing physical exercise. This epidemic of obesity may be one cause for a recent increase in hyperuricemic individuals in Japan. PMID- 18409510 TI - [Urate production and regulation]. AB - The amount of urate depends on the balance between production and excretion. Although only about 10% of hyperuricemia results from overproduction of urate, purine biosynthesis has substantial impacts on the pathogenesis of hyperuricemia. Here, regulation of purine biosynthesis pathway and relevant mechanisms are discussed. Genetic mechanisms for urate overproduction, dietary effects on urate amount and new drug therapy inhibiting urate production are also discussed. PMID- 18409511 TI - [Molecular mechanisms of urate transport in renal tubules: localization and function of urate transporters]. AB - Urate, a naturally occurring product of purine metabolism, is present at higher levels in human blood than in other mammals, because humans have an effective renal urate reabsorption system in addition to their evolutionary loss of hepatic uricase by mutational silencing. The urate transporter URAT1 encoded by SLC22A12 is a urate anion exchanger regulating blood urate levels. URAT1 is localized in the apical membrane of renal proximal tubules and targeted by uricosuric and antiuricosuric agents. Idiopathic renal hypouricemia is due to the genetic defect of SLC22A12. Recently it has been shown that the proximal tubule apical membrane organic anion transporter OAT4 transports urate at low affinity and responsible for the hyperuricemia cased by thiazide diuretics. Transport of urate via URAT1 is driven by the intracellular lactate that is accumulated by Na+/lactate cotransporter slc5a8 and slc5a12. URAT1 is proposed to be involved in the multimolecular complex "transportsome" that allows the cooperation of multiple transporters. PMID- 18409512 TI - [Physiology and dynamics of uric acid in hyperuricemia]. AB - Gout is a type of inflammatory arthritis that is triggered by the crystallization of monosodium urate (MSU) within the joints and is often associated with hyperuricemia. Further understanding the physiology and dynamics of uric acid in human is required to make sure the disease mechanism of both gout and hyperuricemia in clinic. The amount of urate in the body consists urate pool, and which depends on the balance between dietary intake, synthesis, and excretion. Uric acid is a weak acid that exists largely as MSU, the ionized form, in urate pool at physiologic pH. But the solubility of MSU is influenced of pH, temperature and protein of blood and tissue. PMID- 18409513 TI - [Idiopathic hyperuricemia with overproduction of uric acid]. AB - Idiopathic hyperuricemia is generally defined as hyperuricemia caused by unknown origin. Idiopathic hyperuricemia is categorized as overproducing or underexcretion of uric acid. Overproduced uric acid is caused by increased biosynthesis of purine bodies, hypermetabolization of purine bodies, or increased intake of dietary purine bodies. Idiopathic hyperuricemia with overproducing uric acid can be diagnosed by amount of excreted uric acid in the urine. Recently it has been identified that hypertension are frequently associated with myogenic hyperuricemia converted from overproduced hypoxanthine in the skeletal muscles. Some anti-hypertensive drugs including alpha1 blocker, ACE inhibitor, alphabeta blocker, or long-acting Ca blocker attenuated the myogenic hyperuricemia. Thus, these drugs may be helpful in the management of hypertension with hyperuricemia. PMID- 18409514 TI - [Primary hyperuricemia due to decreased renal uric acid excretion]. AB - Hyperuricemia reflects extracellular fluid supersaturation for uric acid. Although dietary, genetic, or disease-related excesses in urate production underlie hyperuricemia in some cases, impaired renal excretion of uric acid is the dominant cause of hyperuricemia. This type of hyperuricemia may be primary (idiopathic) and unassociated with an identifiable disorder. Two important candidates that may affect renal urate excretion were identified recently. One is an organic anion transporter (OAT) family member called urate transporter (URAT) 1. URAT1 has highly specific urate transport activity, exchanging this anion with others including most of the endogenous organic anions and drug anions that are known to affect renal uric acid transport. Another is uromodulin (UMOD), which is the key protein for the pathogenesis of familial juvenile hyperuricemic nephropathy that is characterized by early onset of hyperuricemia and renal failure. The role of these proteins in the cause of hyperuricemia is under investigation. PMID- 18409515 TI - [Familial juvenile hyperuricemic nephropathy (FJHN)]. AB - Familial juvenile hyperuricemic nephropathy (FJHN) is an autosomal-dominant disease characterized by hyperuricemia of underexcretion type, gout, and chronic renal failure. Recent discovery of uromodulin mutations as a cause of FJHN and MCKD2 led a new concept, i.e. uromodulin-associated kidney disease (UAKD). The genotype-phenotype correlation and genetic heterogeneity of FJHN are reviewed. PMID- 18409516 TI - [Deficiencies of hypoxanthine guanine phosphoribosyltransferase (HPRT)]. AB - Inherited mutations of a purine salvage enzyme, hypoxanthine guanine phosphoribosyltransferase (HPRT, EC 2.4.2.8; MIM308000), give rise to Lesch-Nyhan syndrome (MIM300322) or HPRT-related gout called as Kelley-Seegmiller syndrome (MIM300323). In contrast with the most severe phenotype of classical Lesch-Nyhan disease (LND), the least severe phenotype is characterized by hyperuricemia without any neurological or behavioral abnormality, and designated HPRT-related hyperuricemia (HRH). In between these two extremes are phenotypes involving hyperuricemia and varying degrees of neurobehavioral abnormality but without self injury, designated HPRT-related neurological dysfunction (HRND). Marked genetic heterogeneity of HPRT deficiency is well known. More than 300 different mutations in the HPRT gene (HPRT1 which located in Xq26.1), deletion, insertions, duplications, abnormal splicing and point mutations at different sites of the coding region from exons 1 to 9, have been identified. PMID- 18409517 TI - [Increased activity of PRPP synthetase]. AB - PRPP(phosphoribosyl pyrophosphate) synthetase catalyzes the formation of PRPP from ATP and ribose 5-phosphate. Human PRPP synthetase exists as heterogeneous aggregates composed of the 34kDa catalytic subunits (PRSI and PRSII) and other 39kDa and 41kDa components designated PRPP synthetase-associated protein (PAP39 and PAP41). A syndrome of increased activity of PRPP synthetase, an X-linked dominant-inherited disorder, is one of the models of gout caused by increased production of uric acid. By now, around twenty cases have been reported over the world. Two different molecular mechanisms underlie this syndrome: (1) point mutation in the gene coding the primary structure of PRPP synthetase causes the substitution of an amino acid residue and, consequently, the regulatory defects, those are resistant traits to allosteric nucleotide feedback inhibition; (2) increased transcription of PRPP synthetase mRNA causes overproduction of this enzyme protein. The mechanism producing increased mRNA is, however, not elucidated. The Japanese case has been found to be caused by the second mechanism. PMID- 18409518 TI - [Pathophysiology and treatment of secondary hyperuricemia]. AB - Hyperuricemia caused secondly from acquired disorders which affect production or excretion of uric acid is defined as secondary hyperuricemia. Many conditions are associated with this type of hyperuricemia and are classified into three types according to the underlying pathophysiology, including accelerated purine nucleotide degradation, ATP breakdown and purine de novo biosynthesis as overproductive type, use of drugs affecting renal urate handling and renal insufficiency as underexcretion type, or overintake of alcohol or fructose as mixed type. Determining uric acid clearance and urate excretion is important for pointing out original disorder; however, sometimes the result from correcting causal factor should be waited for to fix up a final diagnosis. Anti hyperuricemia agents are used according to the pathophysiology. PMID- 18409519 TI - [Pathological mechanisms of gouty arthritis]. AB - Gout is a disease caused by the deposition of monosodium urate monohydrate (MSU) crystals. Precise mechanisms underlying the initiation of acute gout, however, are not known. Recent investigations provided novel evidence in the pathology of acute gout. A number of studies indicated that MSU crystals can act as a "danger signal" which resembles exogenous adjuvants, and toll-like receptor(TLR)-mediated pathways and/or MyD88-dependent IL-1 receptor pathways are involved in acute gout. Up-regulation of the triggering receptor expressed on myeloid cells 1(TREM 1) in phagocytes by the stimulation with MSU crystals has been demonstrated. Furthermore, pathological significance of NALP 3 inflammasome in gout has been also demonstrated. These findings provide a new insight into the mechanisms underlying the initiation of MSU crystal-induced acute inflammation. PMID- 18409520 TI - [Tophaceous gout]. AB - Identification of tophi indicates a definitive diagnosis of gout. However, recently they are rarely encountered. Tophi are most often seen in tissues that have a poor blood supply and low temperature, such as the ear helix and first MP joint. The nodules are yellowish-white, and non-tender, and range in size from 1 mm to 7 cm. Aspiration yields a chalky-like material that appears as needle-like crystals under light microscopy. In more advanced cases, tophi have a "punched out" appearance with an "overhanging" margin on X-ray images. Differential diagnoses include rheumatoid nodules, xanthoma tuberosa, and CPPD crystal deposition diseases. Tophi respond well to anti-hyperuricemic therapy, during which they gradually decrease in size. However, a huge nodule may need to be surgically removed. PMID- 18409521 TI - [Urolithiasis and nephropathy complicated with gout]. AB - Urolithiasis is a clinically important complication of gout. For effective prevention of this complication, it is necessary to comprehend the factors known to be important in its development. Our analysis of stones in gout patients revealed that the incidence of common calcium salt stones was over 60%, while that of uric acid stones was only about 30%. This implies that the disruption of uric acid metabolism promotes not only uric acid stones but also calcium salt stones. Twenty-four per cent of gout patients showed acidic urine throughout the day. Urinary management, which consists of hydration and alkalization of urine, is indispensable along with control of the serum urate level in the treatment of gout. PMID- 18409522 TI - [How do we set the standard value of serum uric acid levels?]. AB - In most medical facilities in Japan, either uricase-catalase or uricase peroxidase method has been adopted as a sensitive determination of serum uric acid concentration. However, the values obtained from the same patients at different time points are often variable with those methods. Accelerated generation of uric acid and impaired excretion in the kidney are promoted by several dietary factors, such as foods with higher content of sugars (fructose and xylitol), fat and purine bases, and by alcohol consumption, starvation and dehydration. In contrast, hyperglycemia and excess salt ingestion are conductive to accelerate urate excretion. Physicians should notice representative factors fluctuating serum uric acid levels as described above. PMID- 18409523 TI - [Establishment of therapeutic goal and plan of gout and asymptomatic hyperuricemia]. AB - Gout is a crystal deposition disease. European and Japanese guidelines of management for gout recommend that serum urate concentration should be maintained below 6.0 mg/dL to promote crystal dissolution leading to prevention of recurrent gouty attack. Although allopurinol is recommended to be an adequate drug for urate lowering therapy in all gouty patients by European guideline, it is desirable that allopurinol is indicated in patients with overproduction type and benzbromarone in patients with underexcretion type, recommended by Japanese guideline. Asymptomatic hyperuricemia dose not equate to gout. As there is no evidence to support treatment of isolated hyperuricemia with urate lowering therapy currently, it is difficult to establish lowering goal of serum urate level in patients with asymptomatic hyperuricemia. Advice regarding lifestyle and treatment of associated comorbidity should be preferred to urate lowering therapy. However, urate lowering therapy may be indicated in high risk patients with hyperuricemia who are suffered from hypertension, diabetes mellitus, ischemic heart disease and renal insufficiency. PMID- 18409524 TI - [Practical strategies for lifestyle modification in people with hyperuricemia and gout treatment through diet, physical activity, and reduced alcohol consumption]. AB - There has been an explosive increase in the prevalence of hyperuricemia and gout in Japan, suggesting the recent lifestyle change may be a key factor leading to this pathophysiological condition. In addition, people with hyperuricemia are often associated with various morbid conditions constituting the metabolic syndrome, such as abdominal obesity, hypertension, dyslipidemia and impaired glucose tolerance. Therefore, healthy lifestyle interventions would be a basic therapeutic approach not only to hyperuricemia but to metabolic syndrome, though it is not easy to promote behaviour changes. This review focuses on strategies for lifestyle intervention for clinical practice, including how we advise patients on appropriate diets, physical activity and alcoholic beverage consumption. PMID- 18409525 TI - [Uricosuric agent]. AB - Urate lowering treatment is indicated in patients with recurrent acute attacks, tophi, gouty arthropathy, radiographic changes of gout, multiple joint involvement, or associated uric acid nephrolithiasis. Uricosuric agents like benzbromarone and probenecid are very useful to treat hyperuricemia as well as allopurinol (xanthine oxidase inhibitor). Uricosuric agents act the urate lowering effect through blocking the URAT1, an urate transporter, in brush border of renal proximal tubular cells. In order to avoid the nephrotoxicity and urolithiasis due to increasing of urinary urate excretion by using uricosuric agents, the proper urinary tract management (enough urine volume and correction of aciduria) should be performed. PMID- 18409526 TI - [Inhibitors of xanthine oxidoreductase]. AB - Inhibitors of xanthine oxidoreductase decrease production of uric acid, thus they act as hypouricemic drugs. Allopurinol, a prototypical xanthine oxidoreductase inhibitor, has been widely prescribed for treatment of gout and hyperuricemia. However, severe side effects of allopurinol may occur in patients with renal insufficiency. Recently, novel nonpurine selective inhibitors of xanthine oxidoreductase have been developed as potential alternatives to allopurinol. They have different inhibition mechanisms, utilizing the enzyme structure and the reaction mechanism. Such variation of the inhibition mechanism affects/in vivo/hypouricemic effects of the inhibitors. PMID- 18409527 TI - [Other antihyperuricemic agents]. AB - It has been reported that hyperuricemia might be responsible for cardiovascular diseases as well as gout and renal injury. Hypertension and hyperlipidemia, which are also responsible for cardiovascular diseases, are often associated with hyperuricemia. Thus, the treatment of hypertension and hyperlipidemia associated with hyperuricemia is also important. Losartan, an antihypertensive agent, and fenofibrate, an antihyperlipidemic agent, are known to have uric acid lowering effects. Both agents are useful for hyperuricemia with associated with hypertension and hyperlipidemia. In this section, we reported the characteristics and usefulness of these two agents in hyperuricemic patients with hypertension and hyperlipidemia. PMID- 18409528 TI - [New antihyperuricemic medicine: febuxostat, Puricase, etc]. AB - An inhibitor of xanthine dehydrogenase (XDH), allopurinol, and uricosuric agents, probenecid and benzbromarone, have been used for more than 20 years in the treatment of hyperuricemia and gout. However, they are inconvenient in some situations. With regard to allopurinol, the dosage reduction is recommended in patients with renal insufficiency for preventing from rare adverse effect, bone marrow depression. Benzbromarone also has quite rare adverse effect, fulminant hepatitis. Recently several new therapies have been developed such as new XDH inhibitors urate transporter (URAT) 1 inhibitor, and a modified recombinant uricase. The dosage reduction of the new XDH inhibitors, febuxostat and FYX-051, is not necessary in patients with renal insufficiency because renal excretion is not main excretory pathway. JTT-552 is a first medicine targeting on URAT1. Polyethylene glycol (PEG) conjugation with recombinant uricase sufficiently reduces the immunogenicity to permit repeated dosing and the clinical trials are ongoing for patients with treatment-failure gout and hyperuricemia. PMID- 18409529 TI - [Relationship between hyperuricemia and metabolic syndrome]. AB - Metabolic syndrome is a cluster of cardiovascular risk factors such as hypertriglyceridemia, hypertension, insulin resistance, based on visceral fat accumulation. Hyperuricemia is also thought as a one of the complications of metabolic syndrome. Hyperinsulinemia caused by insulin resistance induces the low excretion type hyperuricemia. In contrast visceral fat accumulation itself causes the hypersynthetic type hyperuricemia through increased fatty acid influx into the liver. Recently hyperuricemia is suggested to play a causal role for the metabolic syndrome. Xanthine oxido-reductase, a key enzyme of uric acid metabolism was indicated as one of regulatory factors in adipocyte differentiation. These studies may shed a new light on the understanding of the relationship between hyperuricemia and metabolic syndrome. PMID- 18409530 TI - [AMPD genes and urate metabolism]. AB - AMPD (AMP deaminase), an enzyme catalyzing AMP to IMP, plays an important role in purine/urate metabolism. AMPD is encoded in 3 genes for 3 isozymes M, L and E (H), respectively. In humans, 2 AMPD deficiencies have been reported in skeletal muscles (AMPD1 deficiency) and red blood cells (AMPD3 deficiency). AMPD1 deficiency has been found in patients with metabolic myopathy. AMPD regulates important purine nucleotides including ATP, ADP, AMP, IMP et al. Also, AMPD deficiency may change the level of adenosine, an important bioactive molecule. In addition, AMP activated protein kinase (AMPK) activity, controlled by intracellular AMP, has an important role as an energy sensor. Therefore, AMPD may control the systemic metabolic status by changing AMPK activity through the AMP level. PMID- 18409531 TI - [Uromodulin mutation and hyperuricemia]. AB - The discovery of uromodulin mutations as a cause of FJHN and MCKD2 raises a new question; Why the mutant uromodulin causes uricemic underexcretion and hyperuricemia? Moreover, an old and still unsolved question is now highlighted; What is the physiological function of uromodulin? Recent experimental data on intracellular trafficking of uromodulin mutants and histopathological findings on renal biopsy specimens of patients are introduced. Hypotheses on mechanisms of FJHN/MCKD2 and supporting and contrary experimental findings are reviewed. PMID- 18409532 TI - [Adenine phosphoribosyltransferase deficiency and its purine metabolism]. AB - Adenine phosphoribosyltransferase (APRT) deficiency is a genetic disorder that causes 2,8-dihydroxyadenine (DHA) urolithiasis. Based on the level of residual enzyme activity in cell extracts, two types of APRT deficiency have been identified. Type I is complete enzyme deficiency. Type II shows residual activity in cell lysates, but enzyme activity is not demonstrable in intact cells. Patients with type II have at least one M136T allele and have been identified mainly in Japanese. Thus, in Japanese, patients with type I deficiency are homozygous for APRT*Q0 (null alleles) and patients with type II deficiency are either homozygotes with APRT*J (M136T allele) or compound heterozygotes with APRT*J/APRT*Q0. PMID- 18409533 TI - [New treatment for spasticity--intrathecal baclofen therapy]. AB - Since its introduction in the late 1980s, intrathecal baclofen(ITB) therapy has become the standard treatment for severe generalized spasticity of both spinal and cerebral origin. More than 50,000 pumps were implanted in the world. Long term efficacy of ITB therapy is excellent, although it is frequently associated with complications such as infections, catheter malfunctions, and cerebrospinal fluid leaks. There are some reports of death with withdrawal, overdose and infection. For a good results we need to discuss factors involved in selecting the appropriate treatment modalities for the individual. The treatment of these patients is optimized in the setting of a multidisciplinary team. PMID- 18409534 TI - [Guidelines for the management of Helicobacter pylori--Maastricht III-2005 and Japanese guidelines]. AB - The guidelines on the management of Helicobacter pylori were updated at the European Helicobacter study group third Maastricht consensus conference in March 2005. Especially, this conference emphasis on the management of non ulcer dyspepsia, GERD, and the patients who use non steroidal anti-inflammatory drug. Eradication of H. pylori is recommended in patients with peptic ulcer, low grade MALT lymphoma, atrophic gastritis, unexplained iron deficiency anemia, chronic idiopathic thrombocytopenic purpura and first degree relatives of patients with gastric cancer. H. pylori eradication is less effective than proton pomp inhibitor(PPI) treatment in preventing ulcer recurrence in long term NSAIDs users. This meeting also emphasized on the relationship between H. pylori and gastric cancer. The guideline concluded that H. pylori eradication has the potential to reduce the risk of gastric cancer development. Japanese guideline in 2003 does not mention the effect of eradication for prevention of gastric cancer. The H. pylori eradication and new strategy should be desirable for global strategy of gastric cancer prevention. PMID- 18409535 TI - [Rasmussen encephalitis and non-herpetic acute limbic encephalitis]. AB - Rasmussen syndrome (RS) and non-herpetic acute limbic encephalitis (NHALE) have pathophysiological background related with autoimmunity to glutamate receptors (GluRs) after infections. RS and NHALE were reviewed, depending mainly on our recent studies. RS is the prototype of autoimmune-mediated epilepsy. In patients with RS, several kinds of autoantibodies against neuronal molecules, for example, GluR3, GluRepsilon2 (NMDA-R2B), etc., are reported. These autoantibodies are not specific for RS. About autoantibodies against GluR3, significance and stimulating effects to GluR3 are controversial. Autoantibodies against GluRepsilon2 were detected in all patients within six months from epilepsy onset, and in some patients at chronic stage. These data suggest that autoantibodies against GluRepsilon2 may be involved in the pathological mechanisms in the early stage, but we could not confirm the effect of the autoantibodies from RS patients on excitatory postsynaptic NMDA current using patch clump methods. However, anti double-stranded DNA antibodies in patients with SLE are reported to cross-react with n-terminal of GluRepsilon2, and cause neuronal apoptosis in rat hippocampus, ensuing memory impairment, and emotional behavior impairment in mice. Therefore, autoantibodies against GluRepsilon2 may contribute to the cognitive and behavioral changes in RS. Concerning about cellular immunity in RS, lymphocytes stimulating tests revealed peripheral lymphocytes sensitized by antigens containing GluRepsilon2. Cytotoxic T cells (CTLs) excreting Granzyme B were reported in resected brain tissue, and we confirmed the elevated levels of Granzyme B, not in sera, but in CSF. These data suggest that CTLs activated by infection invade into CNS, and recognize neural antigens, and excrete Granzyme B. The incidence of NHALE is 4.1/1 million/year in Japanese adults. Our study in 91 adult patients with NHALE revealed the following characteristics. Mean onset age was 35.2 +/- 16.9 years old, and preceding infections existed in 68.7% of patients, and predominant symptoms at the onset were psychiatric symptoms (33.3%) and convulsions (25.0%). CSF showed slightly elevated cell counts (55.5 +/- 139.9), protein levels (48.1 +/- 36.0 mg/dl), and IgG levels (4.5 +/- 3.9 mg/dl). MRI lesions with high intensity were found in 40.8% (DWI) and 54.2% (FLAIR) of patients in various stages after onsets. Autoantibodies against GluRepsilon2 in sera were detected in approximately 60% of NHALE patients from acute to chronic stages, and the autoantibodies in CSF were detected in 51.8% (acute stage), 41.4% (recovery stage), 28.6% (chronic stage) of patients and included epitopes to n terminal of GluRepsilon2 (NT1). These data suggest that autoantibodies against GluRepsilon2 produced in sera after infection infiltrate into CNS through damaged BBB in acute stages, and affect n-terminal of GluRepsilon2. In chronic stage, recovery of function of BBB reduces levels of the autoantibodies in CSF. Because BBB in hippocampi and amygdala are vulnerable, autoantibodies against GluRepsilon2 including epitopes to n-terminal may contribute to the limbic symptoms around onset. Among several autoantibodies related with NHALE, autoantibodies against GluRepsilon2 were found in patients around 15-34 years old, autoantibodies against VGKC were around 50.4 years old, autoantibodies against NAE were around 59 years old, autoantibodies against Hu were around 61.5 years old. These data suggest that autoantibodies related with NHALE have age dependent heterogeneity. PMID- 18409536 TI - [Clinical features and course of 5 cases with HIV encephalopathy]. AB - Clinical features and courses of 5 cases with HIV encephalopathy were reported. The subjects were among the patients registered as HIV patients at the Nagoya Medical Center, between 1996 and 2005. There were 458 patients with HIV infection including 127 cases of AIDS. All patients suffered from severe immunological deficiency when HIV encephalopathy developed. Other opportunistic infections had also occurred in three patients. HIV encephalopathy was one of the presenting manifestations of HIV infection in four patients, and no patients had received antiretroviral therapy. HAART improved motor disturbance and their ADL became independent except for one case. Improvements in neuropsychological examination scores were noted in all cases. Recovery from psychiatric symptoms, however, was incomplete. Four patients could not work, and 3 needed psychological treatment due to behavioral abnormalities. HIV encephalopathy is not a lethal disease but the functional prognosis was very poor. New therapy is needed for HIV encephalopathy. PMID- 18409537 TI - [Case of Gerstmann-Straussler-Scheinker syndrome (GSS-P102L) mimicking variant Creurtzfeldt-Jakob disease in clinical manifestation and MRI findings]. AB - We reported a 51-year-old woman with Gerstmann-Straussler-Scheinker syndrome (GSS P102L) manifesting characteristic MRI findings. At the age of 45, She developed gait disturbance with muscle atrophy in the lower limbs and positive plantar flexor sign. Subsequently, sensory disturbance such as refractory pain in the lower limbs and ataxic gait were developed at the age of 49. Following these clinical symptoms, she finally demonstrated rapid progressive cognitive dysfunction. Just after presenting cognitive dysfunction, cranial MRI was performed. Cranial MRI with diffusion-weighted imaging and FLAIR imaging demonstrated abnormal high intensity lesions in the bilateral pulvinar, caudate nuclei and cerebral cortex. The degree of high signal at the pulvinar was less than those of the cortex and caudate nuclei. A proline-for-leucine substitution at codon 102 of the prion protein gene was demonstrated. These results allowed the diagnosis of GSS (P102L). This is a rare case of GSS (P102L) presenting with high intensity lesions in the bilateral pulvinar on MRI. PMID- 18409538 TI - [Syndrome of inappropriate secretion of anti-diuretic hormone associated with limbic encephalitis due to herpes simplex virus infection--a case report]. AB - We report a case of syndrome of inappropriate secretion of anti-diuretic hormone (SIADH) associated with limbic encephalitis. A 79-year-old woman was admitted with a complaint of fever, disturbance of consciousness and generalized seizure. Her conscious level was E1V2M4 by Glasgow coma scale. Physical examination showed generalized seizure, neck stiffness, hyperreflexia and flaccid paralysis in her all extremities, and pathological reflexes. Blood analysis revealed hyponatremia, decrease of plasma osmolarity, spared secretion of urine sodium and increase of ADH, leading to the diagnosis of SIADH. Cerebrosponal fluid examination showed mild pleocytosis, elevated protein, and normal glucose level. Although herpes simplex virus (HSV) DNA was not detected by the polymerase chain reaction method, titers of anti-HSV IgG antibody elevated chronologically. Brain MRI revealed abnormal T2 and FLAIR high intensities in the cingulate gyrus and hippocampus bilaterally. An EEG revealed periodic synchronous discharges predominantly in the frontal areas. Based on the clinical course, laboratory data, MRI and EEG findings, we diagnosed as SIADH associated with acute limbic encephalitis caused by HSV infection. After the fluid restriction and sodium supply, plasma sodium was normalized. Administration of acyclovir and steroid was not so effective, however her condition improved gradually. Several cases of SIADH associated with limbic encephalitis have been reported; however, the pathophysiology is to be clarified. We thought that in the presented case, SIADH was caused by disturbance of the hormonal control at the hypothalamus on the pituitary gland due to the spreading of inflammation from limbic system to these areas. PMID- 18409539 TI - [Case of rheumatoid meningitis: findings on diffusion-weighted image versus FLAIR image]. AB - We report a 63-year-old man with rheumatoid meningitis. At 47-years-old, he developed rheumatoid vasculitis causing arthralgia and skin ulcer. Although the patient had been treated with prednisolone and cyclosporine A, headache and recurrent focal seizures of the right upper limb and generalized seizures developed. Brain magnetic resonance imaging showed high signal intensity lesions on FLAIR MRI and associated abnormal enhancement of the leptomeninges. Part of the lesions also showed patchy high signal intensity on diffusion-weighted imaging (DWI). This features may be useful for differentiating rheumatoid meningitis from subdural empyema, because the extent of the lesions on DWI matches the lesion on FLAIR imaging in patients with subdural empyema. Cerebrospinal fluid analysis revealed monocytic pleocytosis and negative findings for infection or malignancy. After intravenous administration of methylprednisolone (1,000 mg/day for 3 days), the patient showed improvements in headache, cerebrospinal fluid findings and abnormal hyperintensity on DWI. Rheumatoid meningitis is an extremely rare neurological manifestation, but careful attention should be paid even in the inactive stage of rheumatoid arthritis. This disease tends to present with unilateral supratentorial lesions. In this case, serial diffusion-weighted and FLAIR MRI was useful for following the leptomeningeal lesions. PMID- 18409540 TI - [Subacute encephalitis associated with anti-glutamate receptor antibodies: serial studies of MRI, 1H-MRS and SPECT]. AB - A 32-year-old man who had experienced fever and a pulsating headache of the right occipital region for a month and a transient left hemianopia and numbness in the left arm two weeks prior to presentation was admitted to our hospital because of a seizure. Fluid-attenuated inversion recovery and diffusion-weighted magnetic resonance imaging (MRI) showed high-intensity signals, without reduction of apparent diffusion coefficient value, in the right temporo-occipital cortices. Proton MR spectroscopy (1H-MRS) indicated a decrease in N-acetylaspartate, and single-photon emission CT (SPECT) showed hyperperfusion in the right temporo occipital territory. An examination of the cerebrospinal fluid showed an elevation of mononuclear cells and the presence of anti-glutamate epsilon2 receptor antibodies. All abnormalities shown by these imaging techniques were normalized in the clinical course. This report suggests that MRI, 1H-MRS and SPECT studies were useful in understanding the pathogenesis of encephalitis associated with glutamate receptor antibodies. PMID- 18409541 TI - [Case of non-alcoholic pellagra following gastrectomy]. AB - A 67-year-old man was admitted to our hospital in May 2006 because of gait disturbance, delirium and myoclonus along with dermatitis and diarrhea. Those symptoms became worse in 3 months. He had undergone a gastrectomy, including a fundectomy and jejunal pouch interposition, for early gastric cancer at the age of 65 years. He had no habit of drinking alcohol or unbalanced diet. The triad of typical dermatitis, delirium, and diarrhea led to a diagnosis of pellagra, and all the symptoms disappeared after intravenous administration of nicotinate and vitamins. With a gastrectomy, fundectomy performed with jejunal pouch interposition has been regarded as a superior method for postoperative nutrition, but may cause vitamin deficiency. Thus, vitamin deficiency must be considered as a potential cause in neurologic patients who underwent surgical treatment for disorders of digestive tract, regardless of the procedure utilized. PMID- 18409542 TI - [Computed tomography (CT) angiography with multidetector row helical CT for detecting spinal dural arteriovenous fistula]. AB - A 59-year-old man had have dysuria and left thigh pain for 2 months. He also experienced transient weakness of both legs twice, which recovered within a few hours. One month ago, he had bowel disturbance. Lumbar MRI showed a swelling of the conus medullaris. Enlarged and tortuous vessels behind the spinal cord on lumbar MRA and CT myelography was demonstrated. Though angiogram through intercostal and lumbar arteries revealed no abnormal findings, CT angiography with multidetector row helical CT (MDCT) showed abnormal vessel in intradural space at sacral level. Angiogram through left internal iliac artery revealed abnormal vessel fed by branches of the lateral sacral artery at the early arterial phase. Thus, diagnosis of dural arteriovenous fistula (DAVF) at the left S1 level was confirmed. It was indicated that CT angiography with MDCT was useful in detecting spinal DAVF especially at sacral level. PMID- 18409543 TI - [Ictal Gerstmann's syndrome in a patient with symptomatic parietal lobe epilepsy]. AB - A 34-year-old man with astrocytoma in the left parietal lobe had symptomatic partial epilepsy, and he presented transient episodes of acalculia, agraphia and finger agnosia. Occasionally he had difficulty in finding appropriate letters when making an e-mail, and difficulty in writing and calculation. Neurological examinations revealed ictal symptoms of Gerstmann's syndrome without right to left disorientation. No other higher cortical dysfunction or neurological deficits were noted. Scalp EEGs showed frequent, regional ictal discharges in the left parietal area lasting for 60-240 seconds. These clinico-electrographical observations strongly support that epileptic seizures produced a loss of cortical higher function manifesting Gerstmann's syndrome. PMID- 18409545 TI - [Overview: Products of Protein 3000 project]. PMID- 18409544 TI - [Is the presence of anti-glutamate receptor antibody positive evidence for "autoimmune" encephalitis?]. PMID- 18409546 TI - [Intracellular signal transduction]. PMID- 18409547 TI - [Structural proteomics of metabolism-related proteins]. PMID- 18409548 TI - ["Team transcription & translation"aiming at 100 structures]. PMID- 18409549 TI - [Structural genomics on development and differentiation of organisms and replication and repair of DNA]. PMID- 18409550 TI - [Structural and functional proteomics consortium for research on the proteins working in brain and nervous system]. PMID- 18409551 TI - [Protein 3000 project "Transcription and translation"]. PMID- 18409552 TI - [Protein higher-order structure formation system]. PMID- 18409553 TI - [Structural and functional biology for post translational modification and transport in eukaryotes]. PMID- 18409554 TI - [RIKEN structural genomics/proteomics initiative]. PMID- 18409555 TI - [From structures to functions: annotation by structural bioinformatics]. PMID- 18409557 TI - [Science and big stakes: invisible outcome of the "big science"]. PMID- 18409556 TI - [Dichtung und wahrheit]. PMID- 18409558 TI - [Comments on P3000 from an X-ray structural biologist]. PMID- 18409559 TI - [Protein structure initiative]. PMID- 18409560 TI - [Histone turnover in the dynamic regulation of chromatin]. PMID- 18409561 TI - [Strategy of HCV proliferation]. PMID- 18409562 TI - [Clinical usefulness and health-economic benefits of a new sheet-like medical device (SD-101) for the diagnosis of sleep apnea syndrome]. AB - The SD-101 (Kenzmedico co. Ltd., Saitama, Japan), a non-invasive medical device capable of measuring respiratory parameters during sleep, has recently been developed. It operates while placed under the body like a bed pad equipped with 162 pressure sensors, with the patient in bed. To evaluate the efficacy and safety of the SD-101 for the diagnosis of sleep apnea syndrome (SAS), we enrolled 52 patients with suspected SAS (45 men and 7 women; mean age, 45.6 +/- 10.9 years) in this study. Each subject underwent measurement using the SD-101 and a polysomnograph simultaneously, and we analyzed and compared them. In addition, health-economic benefits of the SD-101 were estimated based on the results. A significantly strong correlation was obtained between the apnea hypopnea index of PSG and its of SD-101 (r = 0.86, p < 0.0001). No adverse event due to the SD-101 occurred, while use of the SD-101 greatly reduced "feeling of being constrained" and discomfort during examination (Wilcoxon test: p < 0.0001). These findings could indicate that the SD-101 is clinically useful and will make a contribution to health-economic benefits for SAS in Japan. PMID- 18409563 TI - [Q fever in hospital-acquired pneumonia]. AB - We studied the effects of Q fever in hospital-acquired pneumonia. The subjects consisted of 121 cases with hospital-acquired pneumonia treated during the period from December 2004 till June 2007. Q fever was diagnosed using a PanBio Coxiella burnetii ELISA test kit. There were no patients with acute infection by Coxiella burnetii. It is concluded that C. burnetii cannot induce onset of hospital acquired pneumonia. PMID- 18409564 TI - [A case of chronic empyema caused by Salmonella Livingstone with soft-tissue abscess]. AB - A 65-year-old man with old tuberculous pleuritis was admitted to our hospital because of swelling and heat on the right upper abdomen on December 8, 2006. Computed tomography showed the presence of thickening and calcification of the right pleura, capsular effusion, and soft-tissue swelling of the right upper abdomen. Salmonella Livingstone was isolated from the aspiration specimen of the capsular effusion. Chronic empyema caused by Salmonella Livingstone with soft tissue abscess was diagnosed, and was treated with 400 mg per day levofloxacin. Salmonella spp. is known as a common pathogen of gastroenteritis. Pleuropulmonary Salmonella infection is rare, and only 5 cases have been reported in Japan. However, this organism can cause respiratory tract infection in a patient with an impaired host defence system such as old tuberculosis. PMID- 18409565 TI - [A patient with Mycobacterium avium lung disease presenting with rapid, progressive and multiple cavity formation, who had been treated rheumatoid arthritis with disease modifying anti-rheumatic drugs (DMARDs)]. AB - An 80-year-old woman presented with rapid, progressive and multiple cavitary lesions in both lungs. Rheumatoid arthritis had been diagnosed and been treated with prednisolone (5 mg/day) and bucillamine since 1996. Due to worsening of arthralgia, methotrexate (6 mg/week) and leflunomide (10 mg/day) had been added to the medication since 2003. In April 2005, her chest radiography revealed multiple cavities and nodules predominantly in both upper lung fields, although she complained of no respiratory symptoms. No pathogenic organisms were found, and the cavitary and nodular shadows were increased rapidly within the next 2 months. Therefore, the patient was referred to our hospital in July 2005. Repeat microbiologic findings of sputum were negative for bacteria and fungi, except for Mycobacterium avium (M. avium). She was given a diagnosis of M. avium lung disease, and it seemed to be associated with her compromised status caused by disease modifying anti-rheumatic drugs (DMARDs). She was then successfully treated with combined chemotherapy employed clarithromycin, rifampicin, ethambutol and streptomycin. So far, rapid and progressive deterioration of non tuberculous mycobacterial lung disease accompanied with an intake of DMARDs had not been reported in Japan. An increase of M. avium complex lung disease in the elderly is now becoming a problem among respiratory physicians. This case highlights the fact that patients who are scheduled to be given DMARDs, particularly elderly case, should be considered to be at an elevated risk of developing non-tuberculous mycobacterial (NTM) lung disease, and the risk of NTM infection should be excluded before prescribing drugs. PMID- 18409566 TI - [A case of idiopathic bronchial artery aneurysm connecting to the pulmonary artery]. AB - Case. A 54-year old woman was found to have an abnormal shadow in a regular checkup chest X-ray film. We suspected a bronchial artery aneurysm based on a contrast-enhanced chest CT. Aortography, bronchial arteriography and pulmonary arteriography showed a bronchial artery aneurysm, 2 cm in diameter, connecting the right bronchial artery, inferior phrenic artery, and pulmonary artery. Bronchoscopy revealed dilatation and hypervascularity of moniliform submucosal vessels below the right truncus intermedius. She underwent bronchial artery embolization several times, but new feeding vessels developed each time. We considered this case required surgical resection. Bronchoscopy after aneurysmectomy revealed decrease of moniliform submucosal vessels. Histological examination revealed a three-layer structure leading to both the pulmonary and bronchial arteries. Conclusion. We diagnosed idiopathic bronchial artery aneurysm connecting to a pulmonary artery. PMID- 18409567 TI - [A case of drug-induced interstitial pneumonia caused by TS-1]. AB - A 75-year-old man underwent right hemi-colectomy in 2003. After recurrence in November, 2005, he had been given CDDP and TS-1. He had complained of cough with white sputum and fever from the end of October, 2006, however, antibiotics did not improve his symptoms. The chest CT showed diffuse ground glass opacities (GGO) in bilateral lung fields. Steroid treatment improved his physical condition and chest x-ray. Drug-induced pneumonia caused by TS-1 was diagnosed data from a drug lymphocyte stimulation test (DLST). Drug induced interstitial pneumonia can therefore occur after long-term administration. PMID- 18409568 TI - [A case of small cell lung cancer complicated by Cushing syndrome]. AB - When a 45-year-old man visited his local hospital with a complaint of chest discomfort and coughing, an abnormal chest X-ray shadow was found. On referral to our department, pigmentation was mainly observed on his face, however, no striae cutis distensae or facial swelling was observed. After a systemic examination, Stage IV small cell lung cancer with Cushing's syndrome was diagnosed. Marked increases in ACTH levels were observed, but no abnormality was found in the pituitary MRI findings, and therefore ACTH-producing small cell lung cancer was suspected. Chemotherapy was administered in addition to appropriate treatment for Cushing's syndrome. The tumor temporarily began to decrease in size, but hypercortisolemia remained, and thus adrenal hormone synthesis-inhibiting therapy was concurrently administered. Subsequently, cortisole began to decrease, but it could not be sufficiently controlled, resulting in occasional colonization by bacteria. Hypercortisolemia is believed to induce the suppression of the immune system, thereby affecting the outcome and QOL more than the immediate symptoms of cancer. Therefore, from an early stage, it is believed that adrenal cortical hormone synthesis-inhibiting therapy should be performed concurrently with chemotherapy, if hypercortisolemia cannot be controlled by radiation and chemotherapy alone. PMID- 18409569 TI - [A case of chest pain variant asthma with headache]. AB - A 37-year-old man consulted our hospital because of severe constricting pain at the right side of the chest and head. Since his chest pain and headache improved with inhaled procaterol hydrochloride, chest pain-variant asthma was diagnosed. Not so many articles have been reported that concerned with this disease. There is a need for a better dissemination of knowledge about this disease. PMID- 18409570 TI - [A case of relapsing polychondritis monitored by repeated measurements of flow volume curve]. AB - A 57-year-old man was admitted to our hospital with hoarseness, productive cough, and dyspnea. Chest CT revealed a thickening of the tracheal wall, and the flow volume curve showed a constrictive upper airway flow pattern. Bronchoscopy revealed disappearance of cartilaginous rings and tracheal stenosis. Biopsy of the cricoid cartilage showed findings compatible with relapsing polychondritis and a diagnosis of relapsing polychondritis was made in accordance with Damiani's criteria. The flow-volume curve and symptoms were immediately improved by methylprednisolone pulse therapy, but on reduction of oral prednisorone, recurrence was identified by flow-volume curve and bronchoscopy. Administration of dapson and cyclophosphamide in addition to oral prednisolone after intravenous methylprednisolone and flow-volume curve resulted in improvement. In this case, monitoring of flow-volume curve was useful for the evaluation of airway complication in relapsing polychondritis. PMID- 18409571 TI - [A case of small cell lung carcinoma complicated by Lambert-Eaton myasthenic syndrome]. AB - A 54-year old man was admitted with general fatigue, muscle weakness and dyspnea on effort. Medical examinations led to a diagnosis of small cell lung carcinoma (SCLC) with Lambert-Eaton myasthenic syndrome (LEMS). Marked improvement of SCLC and symptoms of LEMS were recognized twice during chemoradiotherapy. On his third admission, he showed muscle weakness, dysaethesia, and neurodysfunction of the bladder and rectum. We initially considered these symptoms to be due to spinal metastasis because MRI findings showed multiple spinal metastases. However, electoromyogram and nerve conduction study demonstrated that his muscle weakness resulted from LEMS though dysethesia and neurodysfunction of bladder and rectum were caused by spinal metastasis. We believe that it is important to perform electomyogram and nerve conduction studies, not only radiographic findings, to detect the "hidden" symptoms of LEMS. PMID- 18409572 TI - [A case of lung adenocarcinoma effectively treated with sivelestat and corcicosteroid for drug-induced lung disease due to gefitinib]. AB - A 70-year-old Japanese man was re-admitted because of relapse of adenocarcinoma of the lung. He received daily administration of gefitinib as second-line chemotherapy. He was given a diagnosis of drug-induced lung disease due to gefitinib on day 6 because of hypoxemia and ground glass opacities in the bilateral lung fields. There was no response to corticosteroid pulse therapy. Continuous administration of sivelestat was intravenously added from day 9. Although mechanical ventilation was required for 10 days, lung infiltrates and hypoxia gradually improved. Sivelestat and corcicosteroid was apparently effective in this case and may be useful treatment for drug-induced lung disease due to gefitinib. PMID- 18409573 TI - [A case of rheumatoid arthritis complicated with methotrexate-induced pneumonitis and pneumocystis pneumonia]. AB - A 62-year-old woman with rheumatoid arthritis was given 4 mg/body methotrexate (MTX) every week and 5 mg prednisolone every day. She developed a severe cough starting in the evening after starting taking MTX and after a fever of 38 degrees and dyspnea appeared the patient was hospitalized. On admission, chest CT findings showed diffuse ground glass attenuation. Pathological findings of specimens obtained by transbronchial lung biopsy showed alveolitis with epithelioid cell granuloma. As a section of the specimen did not show cyst staining by Grocott stain, MTX-induced pneumonitis was diagnosed. The same day, methylprednisolone pulse therapy was started and trimethoprim-sulfamethoxazole (TMP-SMX) was given simultaneously, while MTX was discontinued. On hospital day 3, subsequent data showed a high serum level of beta-D glucan and a positive PCR result for Pneumocystis jiroveci in bronchoalveolar lavage fluid (BALF). Additional section of the specimen showed eosinophilic foamy areas on HE staining and cysts measuring 8 microm, consistent with the Pneumocystis jiroveci lesions by Grocott stain. We present a case of rheumatoid arthritis complicated by methotrexate-induced pneumonitis in which pneumocystis pneumonia was demonstrated by clinical and pathological findings. PMID- 18409574 TI - [A case of severe bronchial stenosis caused by a vascular ring in an elderly woman]. AB - We report a case of bilateral severe bronchial stenosis which was caused by compression by ascending and descending portions of the aorta. Asthma-like symptoms developed 9 years previously and dilation of the ascending aorta was noted 2 years previously. Bronchial obstruction was suspected because of diminishing breath sounds in the left hemithorax. Investigations suggested existence of a vascular ring consisting of the left aortic arch, right descending aorta and possibly the right ligamentum arteriosum. The patient fell in shock during bronchoscopy. Emergency implantations of bronchial stents in the right bronchus and then the left bronchus were made. Because of the patient's poor lung function, there was an extremely high risk for surgical treatment. Although airway stenosis caused by vascular anomaly in elderly patients is very rare, when asthma-like symptoms developed in patients with a right-sided descending aorta, physicians should consider the possibility of such condition. PMID- 18409575 TI - [An autopsy case of chronic empyema-associated angiosarcoma that presented with a rapidly growing chest wall mass]. AB - A 76-year-old woman with chronic tuberculous empyema presented with bloody sputa and anorexia in April 2005. Chest X-ray and CT scan showed no obvious change compared with previous films. She complained of a left chest pain in June 2005, and chest CT scan showed a tumor shadow enhanced heterogeneously on the left chest wall and chronic empyema. After admission, the chest wall mass grew rapidly. Neither CT- nor sonographically guided biopsy yielded a definitive diagnosis. Dynamic MRI showed a high signal intensity unlike hematoma. She died of respiratory failure 2 months after the onset of her chest pain. Autopsy revealed angiosarcoma. We should always keep in mind the early diagnosis of malignant tumor and tuberculosis in patients presenting with a chest wall mass and constitutional symptoms during follow-up of chronic tuberculous empyema. PMID- 18409576 TI - [Pulmonary metastasis of fibrosarcomatous variant of dermatofibrosarcoma protuberans: case report and review of literature]. AB - A 63-year-old man had undergone excision of a growing mass with a wide margin in the left supraclavicular fossa. A diagnosis of fibrosarcomatous variant of dermatofibrosarcoma protuberans (DFSP-FS) was made. Three years later, an abnormal chest shadow was detected on a medical checkup. Chest computed tomography showed a heterogeneously-enhanced 2-cm coin lesion with a distinct border in the right lower lobe and a 3-mm nodule in the left lower lobe. Transbronchial lung biopsy specimens from the right lung revealed a DFSP pattern. We then performed right basal segmentectomy and partial resection of the left lower lobe. DFSP is a relatively rare skin tumor that is considered to be intermediate malignancy. It frequently recurs locally but rarely has systemic metastasis. However, DFSP-FS, a subtype of DFSP, has an increased likelihood of systemic metastasis. The lung is the most common site of metastasis of DFSP-FS. DFSP-FS sometimes recurs even a long time after excision. Therefore, long-term follow-up, including chest X-ray and CT are important in DFSP-FS patients. PMID- 18409577 TI - [In vivo analysis of a genetically modified adenoviral vector targeted to human CD40 using a novel transient transgenic model]. PMID- 18409578 TI - [Role of oxidative stress in the susceptibility to smoke-induced emphysema]. PMID- 18409579 TI - [The role of CD8+ T cells in the pathogenesis of COPD]. PMID- 18409580 TI - [Approach of surgeons toward residents: can we increase the future pool of qualified surgeons?]. PMID- 18409581 TI - [New diagnostic imaging and navigation surgery]. PMID- 18409582 TI - [New diagnostic imaging for navigation surgery]. AB - Progress in diagnostic computed tomography (CT) and magnetic resonance (MR) imaging has been remarkable. Multidetector-row CT provides thin-slice images through the upper abdomen, multiphase abdominal imaging, and 3D images of high quality including CT angiography and multiplanar reformation. The development of MR units provides diffusion-weighted images for detecting abdominal tumors, and the steady-state coherent echo method can be used for imaging of vessels without using contrast media. The 3D images provided in CT and MR imaging facilitate anatomic understanding of tumors and vessels and are useful for preoperative navigation. However, we must be careful when using 3D images for diagnosis, because the subjectivity of the 3D image creator may affect the results. Therefore the original axial images should also be referred to. PMID- 18409583 TI - [Usefulness of image-navigated surgery in liver surgery]. AB - Previously, computed tomography (CT) yielded cross-sectional images reconstructed from single-slice CT. However, the recently developed multidetector-row spiral CT provides isotropic voxel data sets, giving clear and precise three-dimensional images of the intrahepatic vascular structure. The vascular anatomy of the liver and relationship between liver tumors and intrahepatic vascular structure can thus be determined. We have developed software for an image-navigated surgery system with which vessels supplying blood to tumors and main hepatic vein drainage can be identified in patients preoperatively. Virtual liver resection can then be performed on a computer using this software. This simulation surgery contributes to making subsequent actual hepatic resection safer and less invasive. PMID- 18409584 TI - [Navigation surgery in the biliary surgery and NOTES: carbon dioxide enhanced MDCT cholangiopancreatography and image overlay surgery]. AB - The preoperative image diagnosis in the biliary disease devolved from analog imaging such as PTCD or ERCP to digital data analysis such as MDCT, MRI, DIC-CT, or MRCP. In late years synchronous visualization of the biliary tracts and associated blood vessels was enabled in fusion method or MRCPA. Carbon dioxide enhanced MDCT cholangiopancreatography depicts the biliary and pancreatic duct by the negative contrasting effect of the carbon dioxide, and is ideal for the surgical navigation. Real time correspondence to the operator's demand is important, and multidirectional observation and volume rendering method are effective for the use of it during surgery. The introduction of image analysis software OsiriX improves complexity and image construction time, and it may be practical. Image overlay surgery consisted of augmented reality and mixed reality is developing toward practical use in navigated surgery. In recent years, NOTES (Natural Orifice Translumenal Endoscopic Surgery) was developed, the support system of the navigation of such flexible endoscope is desired earnestly. PMID- 18409585 TI - [Navigation surgery for pancreatic disease using multislice computed tomography]. AB - In navigation surgery, preoperatively acquired image data are used so that surgical instruments can be guided inside the body while their location is displayed on a computer monitor. It is used in cranial nerve and spinal surgery. In the field of abdominal surgery, however, surgical manipulations in the target area cause major changes in the displayed images compared with those obtained preoperatively, and therefore, with the exception of certain organs, navigation surgery is difficult to apply. In general, this type of surgery aims to use intraoperative image information to improve surgical precision, carry out the preoperative plan accurately, and avoid dangerous areas. Three-dimensional images of the vascular architecture obtained with multislice computed tomography (MS-CT) make it possible to visualize arteries, the portal vein, bile duct, and even the pancreatic duct from any angle, which cannot be done with conventional angiography. Accurate positional relationships in the affected region can be determined preoperatively by manipulating multiplanar reconstruction images at a work station. MS-CT is extremely useful in navigation for safe performance of all types of pancreatectomy. PMID- 18409586 TI - [Preoperative diagnosis of lymph node metastases and sentinel node navigation surgery in patients with upper gastrointestinal cancer]. AB - In spite of recent advances in diagnostic tools such as computed tomography, endoscopic ultrasonography, and positron-emission tomography, preoperative diagnosis of lymph node metastases in patients with upper gastrointestinal (GI) cancer has been problematic because of the low sensitivity and accuracy in the detection of micrometastases. To overcome this issue, the sentinel node (SN) concept has attracted attention in recent years and is anticipated to become a novel diagnostic tool for the identification of clinically undetectable lymph node metastases in patients with early upper GI cancer. For early-stage gastric cancer, in which a better prognosis can generally be achieved using conventional surgical approaches, individualized, minimally invasive gastrectomy based on a combination of laparoscpic surgery with SN navigation surgery should be established as the next surgical milestone. Several issues remain to be resolved in laparoscopic gastrectomy with three-dimensional computed tomography navigation. PMID- 18409587 TI - [New preoperative imaging and navigation systems for the treatment of colorectal cancer]. AB - Recently, many new preoperative imaging and navigation systems for the treatment of colorectal cancer have become available based on highly developed imaging hardware, software, and clinical techniques. In this paper, we describe and review the clinical role of these new methods. Computed tomography colonography (3D-CT) is a feasible and accurate method to diagnose the location, degree of invasion, and presence of extracolonic metastases in colorectal cancer. 3D-CT with CT angiography is also helpful to assess the vascular branching anatomy before and during surgery. With the use of polyethylene glycol solution plus contrast medium bowel preparation, 3D-CT shows the total colon without blind spots and good differentiation between polyps and residual fluid because the residual fluid is tagged with contrast medium. Magnetic resonance (MR) colonography and diffusion-weighted MR imaging provide additional preoperative information without radiation exposure. Positron-emission tomography/CT colonography has the potential to become an accepted technique for evaluation of the nonvisualized part of the colon caused by stenosis and it can increase the diagnostic yield of masses and clinically important polyps in this part of the colon. Although further prospective randomized trials are needed, sentinel node mapping and detection of micrometastases may be clinically relevant in colorectal cancer and may improve the selection of patients for chemotherapy. PMID- 18409588 TI - [Current status and future vision of hospital-based cancer registries in Japan]. PMID- 18409589 TI - [The system of Japanese Board of Cancer Therapy]. PMID- 18409590 TI - [What we can learn from a case of medical malpractice--a case of cerebral infarct arising after preventive clipping for cerebral arterial aneurysm]. PMID- 18409591 TI - [Nuss procedure for pectus excavatum in children]. PMID- 18409592 TI - [Stop the "surgeon shortage" what could be done and what should be done right now. Series 3: correction of uneven distribution of doctors]. PMID- 18409593 TI - [Stop the "surgeon shortage" what could be done and what should be done right now. Series 4: reform of national medical budget and expenditure]. PMID- 18409594 TI - Worms bear sludge load. PMID- 18409595 TI - Characterizing airplane plumes on the runway. PMID- 18409596 TI - New photovoltaics change solar costs. PMID- 18409598 TI - Shale-oil development on the fast track. PMID- 18409597 TI - Perchlorate in food. PMID- 18409599 TI - Who will pay for a global climate-technology revolution? PMID- 18409600 TI - Risks of nanotechnology remain uncertain. PMID- 18409601 TI - Why large-scale afforestation efforts in China have failed to solve the desertification problem. PMID- 18409602 TI - Membrane-aerated biofilms for high rate biotreatment: performance appraisal, engineering principles, scale-up, and development requirements. AB - Diffusion of the electron acceptor is the rate controlling step in virtually all biofilm reactors employed for aerobic wastewater treatment. The membrane-aerated biofilm reactor (MABR) is a technology that can deliver oxygen at high rates and transfer efficiencies, thereby enhancing the biofilm activity. This paper provides a comparative performance rate analysis of the MABR in terms of its application for carbonaceous pollutant removal, nitrification/denitrification and xenobiotic biotreatment. We also describe the mechanisms influencing process performance in the MABR and the inter-relationships between these factors. The challenges involved in scaling-up the process are discussed with recommendations for prioritization of research needs. PMID- 18409603 TI - Formation of pesticide nonextractable (bound) residues in soil: magnitude, controlling factors and reversibility. AB - The analysis of the coherent data on nonextractable (bound) residues (NER) from the literature and EU pesticide registration dossiers allows the identification of general trends, in spite of the large variability and heterogeneity of data. About 50% of the pesticides reviewed exhibit a low proportion of NER (less than 30% of the initial amount) while only 12% of pesticides have a proportion of NER exceeding 70%. The lowest proportion of NER was found for dinitroanilines (<20%), and the largest value was obtained for carbamates, and in particular dithiocarbamates. The presence of chemical reactive groups, such as aniline or phenol, tends to yield a larger proportion of NER. NER originating from N heteroatomic ring were found to be lower than those from phenyl-ring structures. Among the environmental factors affecting the formation of NER, microbial activity has a direct and significant effect. Concerning the NER uptake or their bioavailability, consistent data suggest that only a small percentage of the total amounts of NER can be released. The analysis of NER formation kinetics showed that incubation experiments are often stopped too early to allow a correct evaluation of the NER maturation phase. Therefore, there is a need for longer term experiments to evaluate the tail of the NER formation kinetics. Still, the heterogeneity of the NER data between pesticides and for specific pesticides calls for great care in the interpretation of the data and their generalization. PMID- 18409604 TI - Charting a path for innovative toilet technology using multicriteria decision analysis. AB - Practical and theoretically sound methods for analyzing innovative environmental technologies are needed to inform public and private decisions regarding research and development, risk management, and stakeholder communication. By integrating scientific assessments with a characterization of values, multicriteria decision analysis (MCDA) supports the ranking of alternative technology pathways on the basis of technical, financial, and social concerns. We applied MCDAto evaluate the use of NoMix urine separating toilets for managing environmental risk and postponing expensive upgrades to a large wastewater treatment plant near Zurich, Switzerland. Results indicate that, given current priorities, no single, fixed course of action (including the status quo) will be desirable to all stakeholders over the considered time horizon. However, a path forward is suggested that is not significantly disadvantageous to any stakeholder now and leaves open future options, allowing society to achieve overall greater benefits if priorities change, new environmental risks are revealed, or technology improves. While our analysis focuses on a particular catchment in Switzerland, many communities worldwide are faced with an aging and inefficient wastewater treatment infrastructure while also experiencing growth and development. Our framework can help these communities balance the conflicting objectives of diverse stakeholders and gain insight into the role that urine separation can play in transitioning to a more comprehensive and sustainable urban water management system. PMID- 18409606 TI - Near-field commercial aircraft contribution to nitrogen oxides by engine, aircraft type, and airline by individual plume sampling. AB - Nitrogen oxides (NOx) concentrations were measured in individual plumes from aircraft departing on the northern runway at Heathrow Airport in west London. Over a period of four weeks 5618 individual plumes were sampled by a chemiluminescence monitor located 180 m from the runway. Results were processed and matched with detailed aircraft movement and aircraft engine data using chromatographic techniques. Peak concentrations associated with 29 commonly used engines were calculated and found to have a good relationship with N0x emissions taken from the International Civil Aviation Organization (ICAO) databank. However, it is found that engines with higher reported NOx emissions result in proportionately lower NOx concentrations than engines with lower emissions. We show that it is likely that aircraft operational factors such as takeoff weight and aircraftthrust setting have a measurable and important effect on concentrations of N0x. For example, NOx concentrations can differ by up to 41% for aircraft using the same airframe and engine type, while those due to the same engine type in different airframes can differ by 28%. These differences are as great as, if not greater than, the reported differences in NOx emissions between different engine manufacturers for engines used on the same airframe. PMID- 18409605 TI - Bioaccumulation of pharmaceuticals and other anthropogenic waste indicators in earthworms from agricultural soil amended with biosolid or swine manure. AB - Analysis of earthworms offers potential for assessing the transfer of organic anthropogenic waste indicators (AWIs) derived from land-applied biosolid or manure to biota. Earthworms and soil samples were collected from three Midwest agricultural fields to measure the presence and potential for transfer of 77 AWIs from land-applied biosolids and livestock manure to earthworms. The sites consisted of a soybean field with no amendments of human or livestock waste (Site 1), a soybean field amended with biosolids from a municipal wastewater treatment plant (Site 2), and a cornfield amended with swine manure (Site 3). The biosolid applied to Site 2 contained a diverse composition of 28 AWls, reflecting the presence of human-use compounds. The swine manure contained 12 AWls, and was dominated by biogenic sterols. Soil and earthworm samples were collected in the spring (about30 days after soil amendment) and fall (140-155 days after soil amendment) at all field sites. Soils from Site 1 contained 21 AWIs and soil from Sites 2 and 3 contained 19 AWls. The AWI profiles at Sites 2 and 3 generally reflected the relative composition of AWls present in waste material applied. There were 20 AWls detected in earthworms from Site 1 (three compounds exceeding concentrations of 1000 microg/kg), 25 AWls in earthworms from Site 2 (seven compounds exceeding concentrations of 1000 microg/ kg), and 21 AWls in earthworms from Site 3 (five compounds exceeding concentrations of 1000 microg/kg). A number of compounds thatwere present in the earthworm tissue were at concentrations less than reporting levels in the corresponding soil samples. The AWIs detected in earthworm tissue from the three field sites included pharmaceuticals, synthetic fragrances, detergent metabolites, polycyclic aromatic hydrocarbons (PAHs), biogenic sterols, disinfectants, and pesticides, reflecting a wide range of physicochemical properties. For those contaminants detected in earthworm tissue and soil, bioaccumulation factors (BAF) ranged from 0.05 (galaxolide) to 27 (triclosan). This study documents that when AWls are present in source materials that are land applied, such as biosolids and swine manure, AWls can be transferred to earthworms. PMID- 18409607 TI - Commercial aircraft engine emissions characterization of in-use aircraft at Hartsfield-Jackson Atlanta International Airport. AB - The emissions from in-use commercial aircraft engines have been analyzed for selected gas-phase species and particulate characteristics using continuous extractive sampling 1-2 min downwind from operational taxi- and runways at Hartsfield-Jackson Atlanta International Airport. Using the aircraft tail numbers, 376 plumes were associated with specific engine models. In general, for takeoff plumes, the measured NOx emission index is lower (approximately 18%) than that predicted by engine certification data corrected for ambient conditions. These results are an in-service observation of the practice of "reduced thrust takeoff". The CO emission index observed in ground idle plumes was greater (up to 100%) than predicted by engine certification data for the 7% thrust condition. Significant differences are observed in the emissions of black carbon and particle number among different engine models/technologies. The presence of a mode at approximately 65 nm (mobility diameter) associated with takeoff plumes and a smaller mode at approximately 25 nm associated with idle plumes has been observed. An anticorrelation between particle mass loading and particle number concentration is observed. PMID- 18409608 TI - Speciation and chemical evolution of nitrogen oxides in aircraft exhaust near airports. AB - Measurements of nitrogen oxides from a variety of commercial aircraft engines as part of the JETS-APEX2 and APEX3 campaigns show that NOx (NOx [triple bond] NO + NO2) is emitted primarily in the form of NO2 at idle thrust and NO at high thrust. A chemical kinetics combustion model reproduces the observed NO2 and NOx trends with engine power and sheds light on the relevant chemical mechanisms. Experimental evidence is presented of rapid conversion of NO to NO2 in the exhaust plume from engines at low thrust. The rapid conversion and the high NO2/NOx emission ratios observed are unrelated to ozone chemistry. NO2 emissions from a CFM56-3B1 engine account for approximately 25% of the NOx emitted below 3000 feet (916 m) and 50% of NOx emitted below 500 feet (153 m) during a standard ICAO (International Civil Aviation Organization) landing-takeoff cycle. Nitrous acid (HONO) accounts for 0.5% to 7% of NOy emissions from aircraft exhaust depending on thrust and engine type. Implications for photochemistry near airports resulting from aircraft emissions are discussed. PMID- 18409609 TI - Occurrence, phase distribution, and mass loadings of benzothiazoles in riverine runoff of the Pearl River Delta, China. AB - A set of six benzothiazoles was determined in riverine runoff samples of the Pearl River Delta (PRD) collected monthly from March 2005 to February 2006. The concentrations of total benzothiazoles ranged from 220 to 611 ng/L, with benzothiazole (BT) being the most prominent (82%), followed by 2 methylthiobenzothiazole (MBT),thianaphthene (TN), and triphenylene (TP). The annual fluxes ofTN, BT, MBT, dibenzothiophene (DBT), 2-(4 morpholinyl)benzothiazole (24MoBT), and TP from the PRD to the coastal ocean were 1.94, 65.1, 10.1,0.63, 0.18, and 0.89 tons/yr, summing to yield an annual flux of 79 tons/yr for total benzothiazoles. In the PRD, approximately 1.1 x 10(5) tons of rubber are estimated to be released into the environment each year. This corresponds to the annual fluxes of 13 tons/yr for BT and 0.4 tons/yr for 24MoBT from tire particles. The annual fluxes of BT from scrap tires from Japan, Korea, Brazil, the European Union, the United States, and China were 99, 21, 36, 270, 328, and 120 tons/yr, respectively. The fluxes of 24MoBT from the same countries were 3.0, 0.5, 1.1, 8.4, 10.3, and 3.8 tons/ yr, respectively. These results indicated that tire-wear particles and scrap tires are the dominant sources of benzothiazoles in the environment. By comparison, Asia may be the major contributor to the global input of benzothiazoles from auto tires in the coming years. Overall, the six benzothiazoles under investigation appeared to be suitable tracers of pollutant inputs to surface runoff within the PRD aquatic system. In addition, 24MoBT seemed more appropriate than BT to trace tire rubber residues and therefore can be a good indicator of economic development and urbanization in a specific region. PMID- 18409610 TI - Occurrence of organophosphorus flame retardant and plasticizers in three volcanic lakes of central Italy. AB - The concentration levels, distribution, and seasonal fluctuations of 12 organophosphorus flame retardants and plasticizers (OPs), of which some are reported to be toxic to aquatic organisms, were investigated in lakes from June 2006 to June 2007. Three volcanic lakes located in the Lazio area (Central Italy) and characterized by a different anthropical impact were selected. Analysis of lake water samples showed that in closed ecosystems (hydrogeological systems), such as small volcanic lakes, OP contamination may occur even in the absence of industries and treated or untreated waste discharges. The selected substances were found at ng/L concentrations in all lakes. In the two more anthropized lakes tributyl phosphate and tripropyl phosphate were the most abundant OPs, with peaks of respectively 784 and 951 ng/L. Maximum pollution levels were reached in October-November, and concentrations decreased to a minimum value in March-April. Chlorinated OPs showed the same trend, but their concentrations were 1 order of magnitude lower and the level decreasing was shifted with respectto alkyl OPs. On the contrary, tris(2-butoxyethyl) phosphate concentrations were quite similar among all water samples analyzed, indicating that their sources were different in nature. One of the three lakes is an important source of drinkable water, so nine wells situated in its neighborhood were also examined. No correlation between lake water and groundwater contamination could be found. PMID- 18409611 TI - Occurrence and profiles of chlorinated and brominated polycyclic aromatic hydrocarbons in waste incinerators. AB - Chlorinated polycyclic aromatic hydrocarbons (CIPAHs) have been reported to occur in urban air. Nevertheless, sources of CIPAHs in urban air have not been studied, due to the lack of appropriate analytical methods and standards. In this study, we measured concentrations of 20 CIPAHs and 11 brominated PAHs (BrPAHs) in fly ash and bottom ash from 11 municipal/hazardous/industrial waste incinerators, using analytical standards synthesized in our laboratory. Concentrations of total CIPAHs and BrPAHs in ash samples ranged from <0.06 to 6990 ng/g and from <0.14 to 1235 ng/g, respectively. The concentrations of CIPAHs were approximately 100-fold higher than the concentrations of BrPAHs. 6-CIBaP and 1-CIPyr were the dominant compounds in fly ash samples. The profiles of halogenated PAHs were similar to the profiles reported previously for urban air. 1-BrPyr was the predominant BrPAH in fly ash. Concentrations of 6-CIBaP, 9,10-Cl2Phe, 9-CIAnt, and 6-BrBaP in fly ash were significantly correlated with the corresponding parent PAH concentrations. Significant correlation between sigmaCIPAH and sigmaPAH concentrations suggests that direct chlorination of parent PAHs is the mechanism of formation of CIPAHs during incineration of wastes; nevertheless, a comparable correlation was not found for BrPAHs. There was no significant correlation between the capacity and temperature of an incinerator and the concentrations of sigmaCl-/BrPAHs in ash samples, although lower concentrations of all halogenated PAHs were found in stoker-type incinerators than in fixed grate-type incinerators. Toxicity equivalency quotients (TEQs) for CIPAHs in ash samples were calculated with CIPAH potencies. Average TEQ concentrations of CIPAHs in fly ash and bottom ash were15800 pg-TEQ/g and 67 pg-TEQ/g, respectively. Our results suggest that the extent of dioxin-like toxicity contributed by CIPAHs in ash generated during waste incineration is similar to that reported previously for dioxins. Waste incineration is an important source of Cl-/BrPAHs in the urban atmosphere. PMID- 18409612 TI - Influence of nitrate on microbial reduction of pertechnetate. AB - Factors influencing microbial reduction of Tc(VII) in nitrate and radionuclide contaminated aquifer sediments were investigated using sediment microcosms containing organic electron donor, nitrate and Tc(VII). Microcosms underwent nitrate reduction followed by Tc(VII) reduction. During this transition, the microbial community changed from being dominated by bacteria affiliated with the genus Paenibacillus during the nitrate reduction phase, to those affiliated with genera Agrobacterium, Geothrix, and Desulfosporosinus during the Tc(VII) reduction phase. To investigate the mechanism of Tc(VII) reduction, the nitrate reducing strains Agrobacterium FRC-A2, Azoarcus FRC-B1, and a fermentative Clostridium FRC-C11 were isolated from sediment microcosms undergoing Tc(VII) reduction. Nitrate reducing bacteria reduced Tc(VII) effectively only in the presence of Fe(lll) and after nitrate was reduced, implying a major role for Fe(ll) as an electron shuttle in Tc(VII) reduction. It is likely that accumulation of nitrite blocks Fe(ll) production and hence Tc(VII) reduction during the active nitrate reduction phase. The pure culture of Clostridium FRC C11 is able to reduce Tc(VII) enzymatically with H2 or glucose as electron donor and deposits insoluble Tc compounds within the cells in a manner that is not significantly influenced by the presence of nitrate. These results provided a possible mechanism for Tc(VII) reduction independent of Fe(III) and not influenced by nitrate. PMID- 18409613 TI - Role of the air-water interface in the retention of TiO2 nanoparticles in porous media during primary drainage. AB - The increasing use of nanomaterials in commercial products has resulted in increased concerns about their potential environmental impacts. The overall mobility of nanomaterials in the environment may depend in part on their mobility in the unsaturated zone of the subsurface, which may provide a sink for nanomaterials, preventing their spread, or a long-term contaminant source. The objective of this work was to study the dynamic unsaturated transport of titanium dioxide (TiO2) during primary drainage to examine the role of air-water interface formation on nanomaterial retention. A specialized automated system was used to track depletion of TiO2 in the pore solution of a porous medium during dynamic drainage, while simultaneously measuring capillary pressure (Pc) and saturation (S). A continuous mass balance was used to calculate the mass of retained TiO2 nanoparticles. Experiments were specifically designed to minimize TiO2 interactions with solid surfaces to allow direct assessment of the role of the air-water interface. Results indicate that the mass of retained TiO2 increases as saturation decreases at all drainage rates, with slower drainage rates corresponding to greater retention at a given saturation. Normalizing the retained mass (M) bythe measured air-water interfacial area (A) shows near constant M/A values at high saturations (S > 0.4) and increasing M/A values with decreasing saturation as saturation drops below 0.4. This result may indicate air water interfacial adsorption at high saturations, with increasing contributions from film straining at lower saturations. PMID- 18409614 TI - Adsorption thermodynamics of p-arsanilic acid on iron (oxyhydr)oxides: in-situ ATR-FTIR studies. AB - The organoarsenical p-arsanilic acid (p-AsA) is used in the U.S. poultry industry as a feed additive and its structure resembles one of the stable biodegradation products of Roxarsone (ROX) in anaerobic environments. With the implementation of recent EPA MCL of total arsenic in drinking water (10 ppb), thereareconcernsaboutthefate of organoarsenicals introduced to the environment through the application of arsenic-contaminated manure. We report herein, for the first time, the thermodynamics of p-AsA binding to Fe-(oxyhydr)oxides using ATR FTIR. ATR-FTIR spectra were used to quantify surface coverage of p-AsA, p AsA(ads), by analyzing the broadband assigned to v(As-O) at 837 cm(-1). Adsorption isotherms were measured in situ at 298 K and pH 7 in the concentration range 1 microM to 40 mM. Values of Keq were obtained from Langmuir model fits and they range from 1411 to 3228 M(-1). We also determined the maximum adsorption capacities of Fe-(oxyhydr)oxides to p-AsA, and they range from 1.9 x 10(13) to 2.6 x 10(13) molecules/cm2. Our results suggest that p-AsA is more mobile than methylated and inorganic forms of arsenic and that the transport of nanoparticles with p-AsA(ads) might play a role in its mobility in geochemical environments. PMID- 18409615 TI - Air-water gas exchange of organochlorine pesticides in Taihu Lake, China. AB - Previous research in the Taihu Lake Region (TLR) of China found high levels of atmospheric organochlorine pesticides (OCPs). To understand the sources and the environmental behaviors of these OCPs in the TLR, research on air-water gas exchange was performed in 2004. Hexachlorocyclohexanes (HCHs), DDT related compounds (DDTs), cis-chlordane (CC), trans-chlordane (TC), heptachlor (HEPT), and alpha-endosulfan in both air and water samples were analyzed, and air-water gas exchange fluxes of these compounds were calculated. The net volatilization flux of alpha-HCH was 58 ng m(-2) day(-1), suggesting that the residue of technical HCH in the lake sediment might have been an important source of alpha HCH to the air of this region after the ban of technical HCH two decades ago. The main components of technical chlordane, TC, CC, and HEPT, each had net volatilization fluxes >230 ng m(-2) day(-1), suggesting that waste discharge from manufacturing plants in the upper region was the main source of chlordane to the lake. Unlike alpha-HCH and chlordane, o,p'-DDT and alpha-endosulfan had net deposition fluxes, suggesting that these compounds were transported through the atmosphere from land sources and then deposited into the lake. The correlation between air concentrations and ambient air temperature indicated that the current sources of o,p'-DDT and alpha-endosulfan were from land; alpha-HCH and chlordane were mainly from the lake. PMID- 18409616 TI - A photochemical source of methyl chloride in saline waters. AB - It is shown experimentallythatthe methoxy group in simple lignin-like molecules can be the source of the methyl group in CH3Cl produced by a photochemical reaction in an aqueous solution of chloride. Terrestrially derived colored dissolved organic matter (CDOM) in river water also yields CH3Cl through a photochemical process in a chloride solution. CDOM extracted from subsurface ocean waters showed some ability to enhance photochemical production of CH3Cl while CDOM from surface water showed no effect. Reactions of the kind described in this paper may be contributors to the marine source of methyl chloride and possibly other alkyl halides. PMID- 18409617 TI - Assessing the impact of nanomaterials on anaerobic microbial communities. AB - As the technological benefits of nanotechnology begin to rapidly move from laboratory to large-scale industrial application, release of nanomaterials to the environment is inevitable. Little is known about the fate and effects of nanomaterials in nature. Major environmental receptors of nanomaterials will be soil, sediment, and biosolids from wastewater treatment. Analysis of anaerobic microbial activity and communities provides needed information about the effects of nanoparticles in certain environments. In this study, biosolids from anaerobic wastewater treatment sludge were exposed to fullerene (C60) in order to model an environmentally relevant discharge scenario. Activity was assessed by monitoring production of CO2 and CH4. Changes in community structure were monitored by denaturing gradient gel electrophoresis (DGGE), using primer sets targeting the small subunit rRNA genes of Bacteria, Archaea, and Eukarya. Findings suggest that C60 fullerenes have no significant effect on the anaerobic community over an exposure period of a few months. This conclusion is based on the absence of toxicity indicated by no change in methanogenesis relative to untreated reference samples. DGGE results show no evidence of substantial community shifts due to treatment with C60, in any subset of the microbial community. PMID- 18409618 TI - Unraveling heavy oil desulfurization chemistry: targeting clean fuels. AB - The sulfur removal chemistry of heavy oils has been unraveled by systematically investigating several heavy oils with an extremely wide range of properties. The heavy oil feed and product properties have been characterized by advanced analytical methods, and these properties have been related to the sulfur conversion data observed in pilot hydrotreating units. These studies coupled with kinetic treatment of the data have revealed that the desulfurization chemistry of heavy oils is essentially controlled by the strongly inhibiting three and larger ring aromatic hydrocarbon content and surprisingly not by the content of the "hard-to-remove" sulfur compounds. Such enhanced understanding of the heavy oil sulfur removal is expected to open new avenues for catalyst/process optimization for heavy oil desulfurization and thereby assist the efficent production of clean transporation fuels. PMID- 18409619 TI - Changes in optical properties caused by UV-irradiation of aquatic humic substances from the amazon river basin: seasonal variability evaluation. AB - Aquatic humic substances (AHS) isolated from two characteristic seasons of the Negro river, winter and summer corresponding to floody and dry periods, were structurally characterized by 13C nuclear magnetic ressonance. Subsequently, AHS aqueous solutions were irradiated with a polychromatic lamp (290-475 nm) and monitored by its total organic carbon (TOC) content, ultraviolet-visible (UV-vis) absorbance, fluorescence, and Fourier transformed infrared spectroscopy (FTIR). As a result, a photobleaching up to 80% after irradiation of 48 h was observed. Conformational rearrangements and formation of low molecular complexity structures were formed during the irradiation, as deduced from the pH decrement and the fluorescence shifting to lower wavelengths. Additionally a significant mineralization with the formation of CO2, CO, and inorganic carbon compounds was registered, as assumed by TOC losses of up to 70%. The differences in photodegradation between samples expressed by photobleaching efficiency were enhanced in the summer sample and related to its elevated aromatic content. Aromatic structures are assumed to have high autosensitization capacity effects mediated by the free radical generation from quinone and phenolic moieties. PMID- 18409620 TI - Mercury mass budget estimates and cycling seasonality in the Florida Everglades. AB - We estimated the mass budget for mercury (Hg) seasonally deposited into the Florida Everglades and investigated seasonality of Hg cycling by analyzing data obtained for water, soil, flocculent detrital material (floc), periphyton, and mosquitofish collected throughout the Everglades freshwater marshes in the 2005 dry and wet seasons. Higher wet season total Hg (THg) in soil, floc, and periphyton agreed with greater Hg amounts entering these compartments during the wet season, probably owing to substantially greater Hg deposition in the wet season than in the dry season. Seasonal differences were absent for THg in surface water. Methylmercury (MeHg) showed mixed seasonal patterns, with higher water and soil MeHg and lower periphyton MeHg in the dry season but no seasonality for floc MeHg. Seasonal variations in Hg deposition, MeHg production and transport, and mass of ecosystem compartments could be responsible for the seasonality of MeHg cycling. Higher mosquitofish THg, higher bioaccumulation factors, and higher biomagnification factors from periphyton to mosquitofish were observed in the wet season than in the dry season, indicating that the wet season is more favorable for Hg bioaccumulation. The mass budget estimation agreed with this result. PMID- 18409621 TI - Sorption of phenanthrene by nonhydrolyzable organic matter from different size sediments. AB - Nonhydrolyzable organic carbon (NHC) and sorption isotherms of phenanthrene (Phen) on six size-fractionated NHC fractions in two sediments from the Pearl River and Estuary, South China, were investigated. It was found that NHC including ancient organic carbon, black carbon, resistant aquatic organic carbon, and aged soil organic carbon consists mainly of aliphatic and aromatic carbon using 13C nuclear magnetic resonance spectroscopy. The sorption isotherms of Phen by the size-fractionated NHC fractions are nonlinear and are well-fitted to the Freundlich model. For the estuary sediment, the NHC contents and the organic carbon-normalized distribution coefficients (Koc) in the size fractions increase with decreasing particle size. The clay NHC fraction contributes to 70% of the Phen sorption by the bulk NHC isolate. However, for the contaminated river sediment, the NHC contents and the Koc values exhibit no regular variations among the size fractions. The Phen sorption capacities on the size-fractionated NHC fractions of the two sediments are significantly related to their H/C ratios and aliphatic carbon, but negatively to aromatic carbon. The fine-particle NHC fractions with high aliphatic carbon and H/C ratio play a very important role in the sorption, transport, and fate of Phen by the investigated sediments. PMID- 18409622 TI - Biotransformation enzymes and thyroid axis disruption in juvenile rainbow trout (Oncorhynchus mykiss) exposed to hexabromocyclododecane diastereoisomers. AB - Juvenile rainbow trout (Oncorhynchus mykiss) were fed either a reference diet or one of three diets enriched with alpha, beta, or gamma diastereoisomers of hexabromocyclododecane (HBCD, C12H18Br6) for 56 days. This exposure period was followed by 112 days during which all fish were fed the reference diet. Potential effects of HBCD on phase I and II biotransformation enzyme activities and thyroid axis disruption were examined. Disruption of the thyroid axis was most evident in the gamma-HBCD exposed group, as indicated by lower circular FT4 and higher FT3 as well as an increase in thyroid epithelial cell height. However, fish fed the alpha-HBCD enriched diet also exhibited altered glucuronyltransferase activity and thyroid epithelial cell heights and the beta-HBCD group had altered FT4 and FT3 and glucuronyltransferase activity. T4ORD activity was not affected after 14 days, but was significantly lower among all HBCD exposed fish compared to the reference fish after 56 days. Results from these experiments indicate that all isomers have the potential to disrupt thyroid homeostasis. PMID- 18409623 TI - Effect of saline waste solution infiltration rates on uranium retention and spatial distribution in Hanford sediments. AB - The accidental overfilling of waste liquid from tank BX-102 at the Hanford Site in 1951 put about 10 t of U(VI) into the vadose zone. In order to understand the dominant geochemical reactions and transport processes that occurred during the initial infiltration and to help understand current spatial distribution, we simulated the waste liquid spilling event in laboratory sediment columns using synthesized metal waste solution. We found that, as the plume propagated through sediments, pH decreased greatly (as much as 4 units) at the moving plume front. Infiltration flow rates strongly affect U behavior. Slower flow rates resulted in higher sediment-associated U concentrations, and higher flow rates (> or =5 cm/day) permitted practically unretarded U transport. Therefore, given the very high Ksat of most of Hanford formation, the low permeability zones within the sediment could have been most important in retaining high concentrations of U during initial release into the vadose zone. Massive amount of colloids, including U-colloids, formed at the plume fronts. Total U concentrations (aqueous and colloid) within plume fronts exceeded the source concentration by up to 5 fold. Uranium colloid formation and accumulation at the neutralized plume front could be one mechanism responsible for highly heterogeneous U distribution observed in the contaminated Hanford vadose zone. PMID- 18409624 TI - Formation of aqueous MgUO2(CO3)3(2-) complex and uranium anion exchange mechanism onto an exchange resin. AB - The formation of and stability constants for aqueous Mg-UO2-CO3 complexes were determined using an anion exchange method. Magnesium concentration was varied (up to 20 mmol/L) at constant ionic strength (I = 0.101, 0.202, 0.304, 0.406, and 0.509 mol/kg NaNO3), pH 8.1, total [U(VI)] = 10.4 micromol/L under equilibrium with atmospheric CO2. The results indicate that only the MgUO2(CO3)3(2-) complex is formed. The cumulative formation constant extrapolated to zero ionic strength is similar regardless of the activity correction convention used: logbeta113(0) = 25.8 +/- 0.5 using the Davies equation and = 25.02 +/- 0.08 using specific ion interaction theory (SIT). Uranium sorption onto the exchange resin decreased in the presence of Mg putatively due to the formation of MgUO2(CO3)3(2-) that had a lower affinity for the resin than UO2(CO3)3(4-). Uranium sorption results are consistent with an equivalent anion exchange reaction between NO3- and UO2(CO3)3(4-) species to retain charge neutrality regardless of Mg concentration. No Mg was associated with the anion exchange resin indicating that the MgUO2(CO3)3(2-) complex did not sorb. PMID- 18409625 TI - Selenite reduction by mackinawite, magnetite and siderite: XAS characterization of nanosized redox products. AB - Suboxic soils and sediments often contain the Fe(II)-bearing minerals mackinawite (FeS), siderite (FeCO3) or magnetite (FesO4), which should be able to reduce aqueous selenite, thereby forming solids of low solubility. While the reduction of selenate or selenite to Se(O) by green rust, pyrite and by Fe2+ sorbed to montmorillonite is a slow (weeks), kinetically limited redox reaction as demonstrated earlier, we show here that selenite is rapidly reduced within one day by nanoparticulate mackinawite and magnetite, while only one third of selenite is reduced by micrometer-sized siderite. Depending on Fe(II)-bearing phase and pH, we observed four different reaction products, red and gray elemental Se, and two iron selenides with structures similar to Fe7Se8 and FeSe. The thermodynamically most stable iron selenide, ferroselite (FeSe2), was not observed. The local structures of the reaction products suggest formation of nanoscale clusters, which may be prone to colloid-facilitated transport, and may have a higher than expected solubility. PMID- 18409626 TI - Environmental influences on the partitioning and diffusion of hydrophobic organic contaminants in microbial biofilms. AB - A biofilm reactor was used to investigate kinetic and thermodynamic aspects of the sorption of polycyclic aromatic hydrocarbons (PAH) as model compounds for hydrophobic organic contaminants (HOC) to intact microbial biofilms. Effective diffusion coefficients are in the range of 10(-10) cm2 x s(-1) resulting in equilibration times of more than 3 days for a biofilm of 100 microm thickness. Diffusion in the biofilm was strongly temperature-dependent and increased by a factor of 3 (phenanthrene) to 6 (fluoranthene, pyrene) between 5 and 35 degrees C. Drying and rewetting of the biofilm as well as the inclusion of Ca2+ ions and of humic acids all strengthened the biofilm rigidity and slowed down the diffusion of PAH. The later two factors also influenced the thermodynamics of the process as they supported the partitioning of PAH into the biofilm. Humic acid inclusion from solution into the biofilm illustrates that a microbial biofilm can act as a primer allowing for the buildup of a particulate organic phase from dissolved organic matter. PAH metabolites (3-hydroxy-phenanthrene and 1-hydroxy-2 naphthoic acid) showed lower partition coefficients as compared to their parent compounds and 3-hydroxy-phenanthrene also showed a higher diffusion constant, indicating that these transformation products would be easily released into the water phase upon formation during PAH biodegradation in a biofilm. These results allow the quantification of the influence of environmental conditions on a biofilm's function as a sink or as a diffusion barrier for PAH from aqueous solution, and they indicate the importance of kinetic aspects of this partitioning process. PMID- 18409627 TI - Substituent effects on nitrogen isotope fractionation during abiotic reduction of nitroaromatic compounds. AB - Compound-specific analysis of nitrogen isotope fractionation is an important tool for assessing transformation pathways of N-containing organic contaminants. We investigated 15N-fractionation during the abiotic reduction of a series of nitroaromatic compounds (NACs) with intrinsic reactivities covering almost 6 orders of magnitude to evaluate substituent effects on 15N kinetic isotope effects, KIEN. Insights into reaction mechanisms and isotopic elementary reactions of NAC reduction were obtained from comparison of experimental results to density-functional theory (DFT) calculations of intrinsic KIEN. Apparent KIEN values for reduction of NACs by structural Fe(II) in octahedral layers of an iron rich clay mineral were substantial (average +la of 1.038 +/- 0.003), independent of the NACs' reactivity and ring substituent, and larger than reported previously for reduction by Fe(II) species bound to Fe(III)(oxy)hydroxides and mercaptojuglone species (1.031 +/- 0.002). DFT-calculations accounting for semiclassical contributions and quantum-mechanical tunneling yielded a KIEN for N O bond cleavage between 1.031 and 1.041, showed no substituent effect, and thus agreed well with experimental observations. Calculated transition-state structures of NAC reduction intermediates were consistent with H2O elimination from substituted N,N-dihydroxyanilines as the predominant 15N-fractionating elementary reaction. The absence of substituent effects on the apparent KIEN of NAC reduction may simplify the practical application of 15N-fractionation data for the quantification of contaminant transformation in the environment. PMID- 18409628 TI - Identification of significant factors in reburning with coal volatiles. AB - In laboratory-scale flow reactor studies, fuel staging using coal volatiles as a reburning fuel has given superior NOx reduction performance compared to reburning using natural gas. This superior performance of coal volatiles may be caused by an increased yield of hydrocarbon radicals and free nitrogen species in the reburning zone. In this study, kinetic models were used to predict the composition of coal volatiles used in reburning. The reburning process was then examined using a set of designed experiments to examine the components of the coal volatiles and their effects on NOx reduction. Within the concentration ranges studied, reburning temperature and air concentration were shown to control NOx reduction, and reburn gas component concentrations had only minor effects. PMID- 18409629 TI - Optimization of surfactant-enhanced aquifer remediation for a laboratory BTEX system under parameter uncertainty. AB - This study develops a nonlinear chance-constrained programming (NCCP) model for optimizing surfactant-enhanced aquifer remediation (SEAR) processes. The model can not only address the parameter uncertainty, but provide a reliability level for the identified optimal remediation strategy. To solve the NCCP model, stepwise cluster analysis (SCA) is used to create a set of proxy simulators for quantifying the relationships between operating conditions (i.e., pumping rate) and probabilities of benzene levels in violation of standard. Compared to conventional parametric inference techniques, SCA is independent of prior assumptions for model forms (e.g., linear or exponential ones) and capable of reflecting complex nonlinear relationships between operating conditions and probabilities. To alleviate the computational efforts in the optimization process, the generated proxy simulators are repeatedly called by simulated annealing (SA) to test the feasibility of each potential solution. The implicit of the optimal NCCP solutions is discussed through a laboratory-scale SEAR system where porosity and intrinsic permeability are treated as stochastic parameters. It is observed that well locations, environmental standards, reliability levels and remediation durations would have significant effects on optimal SEAR strategies. By comparing the predicted benzene concentration without and with remediation actions, it is indicated that the optimal SEAR process can guarantee the benzene concentration to meet the environmental standard with a high reliability level. PMID- 18409631 TI - Metal flux and dynamic speciation at (bio)interfaces. Part III: MHEDYN, a general code for metal flux computation; application to simple and fulvic complexants. AB - Metal flux at consuming interfaces (e.g., sensors or microorganisms) is simulated in environmental multiligand systems using a new numerical code, MHEDYN (Multispecies HEterogeneous DYNamics), based on the lattice Boltzmann method. The attention is focused on the computation of the maximum flux of Cu(II), that is, the flux controlled by diffusion-reaction in solution, irrespective of processes occurring at the interface. In parts III and IV of this series, three types of typical environmental complexants are studied: (a) simple ligands (OH- and C03(2 )), (b) fulvic or humic substances including many sites with broadly varying rate constants, and (c) aggregates including a broad range of sizes and diffusion coefficients. Part III focuses on computations in the presence of simple ligands and fulvic/humic substances separately, and part IV discusses the case of aggregate complexes alone and the mixtures of all ligands in typical natural waters. These papers describe the dynamic contribution of the various types of sites for fulvic and aggregate Cu(II) complexes for the first time. Whenever possible, the metal fluxes computed by MHEDYN are compared with those given by another code, FLUXY, based on a fully different mathematical approach, and very good agreement between these codes is obtained. In all cases, MHEDYN computes the concentration profile of each complex and its time evolution, as well as the steady-state flux and the corresponding contribution of each complex to the flux. The metal fluxes can be computed at a planar consuming surface such as an organism or a sensor surface, in presence of an unlimited number of complexation reactions of the metal M, and for any metal/ligand concentration ratio, with values of the physicochemical parameters ranging over many orders of magnitude. PMID- 18409630 TI - Understanding trichloroethylene chemisorption to iron surfaces using density functional theory. AB - This research investigated the thermodynamic favorability and resulting structures for chemical adsorption of trichloroethylene (TCE) to metallic iron using periodic density functional theory (DFT). Three initial TCE positions having the plane defined by HCC atoms parallel to the iron surface resulted in formation of three different chemisorption complexes between carbon atoms in TCE and the iron surface. The Cl-bridge initial configuration with the HCC plane of TCE perpendicular to the iron surface did not result in C-Fe bond formation. The most energetically favorable complex formed at the C-bridge site where the initial configuration had the C=C bond in TCE at a bridge site between adjacent iron atoms. In the C-bridge complex, one C atom formed two a bonds to different Fe atoms, while the second C atom formed a sigma bond with a second Fe atom. Surface complexation atthe C-bridge site resulted in scission of all three C-Cl bonds and also resulted in a shortening of the C==C bond to a distance intermediate between a double and a triple bond. Initial configurations with the C==C bond adsorbed at top or hollow sites on the iron surface resulted in formation of C-Fe a bonds between a single C and two adjacent Fe atoms, and the scission of only two C==Cl bonds. Bond angles and bond lengths indicated that there were no changes in bond order of the C==C bond for top and hollow adsorption. Chemisorption at the C-bridge site had an activation energy of 49 kJ/mol and an early transition state where all three C-CI bonds were activated. The early transition state and the loss of all three Cl atoms upon chemisorption are consistent with most experimental observations that TCE undergoes complete dechlorination in one interaction with the iron surface. The absence of chemisorption and scission of only two C--Cl bonds at the Cl-bridge site is consistent with experimental observations that trace amounts of chloroacetylene may also be produced from reactions of TCE with iron. PMID- 18409632 TI - Metal flux and dynamic speciation at (bio)interfaces. Part IV: MHEDYN, a general code for metal flux computation; application to particulate complexants and their mixtures with the other natural ligands. AB - Metal flux at consuming interfaces (e.g., sensors or microorganisms) is simulated in environmental multiligand systems using a new numerical code, MHEDYN (Multispecies HEterogeneous DYNamics), based on the lattice Boltzmann method. The attention is focused on the computation of the maximum flux (i.e.,the flux controlled by diffusion-reaction in solution) of Cu(II). Part III described flux computation in the presence of simple ligands and fulvic/humic substances. This paper (Part IV) discusses the case of metal complexes formed with aggregates including a broad range of sizes and diffusion coefficients and their mixture with simple and fulvic ligands under typical natural water conditions. This paper describes the dynamic contribution of the various size classes of aggregate Cu(II) complexes for the first time. In two typical waters containing mixtures of ligands, the contribution of aggregates is found to be small, whereas that of fulvics may play a major role, even under pH conditions where the lability of their Cu(II) complexes is low. These results point out the great usefulness of MHEDYN for dynamic speciation in very complex mixtures. In all cases, MHEDYN enables us to compute the concentration profile of each complex and itstime evolution, as well as the steady-state flux and the corresponding contribution of each complex to the flux. Thus, MHEDYN should be very useful for comparing theoretical predictions with experimental measurements of metal bioavailability or of dynamic sensor response in a complete aquatic medium. PMID- 18409634 TI - Evaluation of laser absorption spectroscopic techniques for eddy covariance flux measurements of ammonia. AB - An intercomparison was made between eddy covariance flux measurements of ammonia by a quantum cascade laser absorption spectrometer (QCLAS) and a lead-salt tunable diode laser absorption spectrometer (TDLAS). The measurements took place in September 2004 and again in April 2005 over a managed grassland site in Southern Scotland, U.K. These were also compared with a flux estimate derived from an "Ammonia Measurement by ANnular Denuder with online Analysis" (AMANDA), using the aerodynamic gradient method (AGM). The concentration and flux measurements from the QCLAS correlated well with those of the TDLAS and the AGM systems when emissions were high, following slurry application to the field. Both the QCLAS and TDLAS, however, underestimated the flux when compared with the AMANDA system, by 64%. A flux loss of 41% due to chemical reaction of ammonia in the QCLAS (and 37% in the TDLAS) sample tube walls was identified and characterized using laboratory tests but did not fully accountforthis difference. Recognizing these uncertainties, the agreement between the systems was nevertheless very close (R2 = 0.95 between the QCLAS and the TDLAS; R2 = 0.84 between the QCLAS and the AMANDA) demonstrating the suitability of the laser absorption methods for quantifying the temporal dynamics of ammonia fluxes. PMID- 18409633 TI - Experimental determination of LSER parameters for a set of 76 diverse pesticides and pharmaceuticals. AB - Linear solvation energy relationships (LSERs) have more recently been proposed as the method of choice to describe and/or predict the partitioning behavior of neutral organic compounds over a large range of environmental matrices and for a broad variety of compounds in a consistent manner. However, when dealing with more complex, polar compounds with multiple functional groups, it has also been noted that there is a severe lack of substance descriptors quantifying the different intermolecular interactions that these compounds may undergo. In this study, we used a system of eight reversed phase, normal phase, and hydrophilic interaction HPLC systems to determine the substance descriptors for H-bond donor (A) and acceptor (B) interactions and for polarizability and dipolarity (S) for a set of 76 complex compounds containing multiple functional groups, comprising mainly pesticides and some pharmaceuticals. The obtained substance descriptors for most compounds are unique in that values of A, S, and B are high and lie at the very upper end of the numerical range of currently known substance descriptors. The substance descriptors have been cross-compared against literature values of the octanol-water (Kow) and air-water (Kaw) partition coefficients and against a set of heptan-methanol partition coefficients (Khm) experimentally determined with a consistent methodology herein. The comparison confirmed plausibility of the substance descriptors of all except six tested compounds. The newly determined substance descriptors promise to be highly valuable in chemicalfate modeling, allowing, in conjunction with available phase descriptors, for a better representation of partitioning of polar compounds in those models. The results also reveal a systematic deviation of the log Kow values predicted with our substance descriptors from the literature values. The deviation points toward a possible problem when existing LSER equations are applied to polar, multifunctional compounds with high values of A, S, and B. Hence, the substance descriptors determined herein should also be helpful in revisiting the validity of existing LSERs for complex, polar compounds. PMID- 18409635 TI - Phage-borne peptidomimetics accelerate the development of polyclonal antibody based heterologous immunoassays for the detection of pesticide metabolites. AB - Competitive immunoassays for the detection of small analytes, such as pesticides and their metabolites, use haptens that compete with the target compounds for binding to the antibody. This competing hapten can be either the same as the immunizing hapten (homologous assay) or structurally modified mimics of the immunizing hapten (heterologous assay). Polyclonal antibody-based heterologous immunoassays have shown superior sensitivities to homologous ones, butthe synthesis of heterologous haptens may be time-consuming, requiring expertise in synthetic chemistry. In this work we demonstrate that phage display peptide libraries can be used as a source of phage-borne peptidomimetics to facilitate the development of sensitive heterologous assays. Different strategies for the isolation of these peptides were explored using two metabolites of pyrethroid insecticides. The sensitivities of the best competitive phage heterologous enzyme linked immunosorbent assays were 13 fold and 100 fold better than the homologous assay, for the glycine conjugate of trans-3-(2,2-dichlorovinyl)-2,2 dimethylcyclopropane-1-carboxylic acid and 3-phenoxybenzoic acid, respectively. The phage particles were highly versatile as tracer reagents, allowing the use of enzymatic, chemiluminescent, or immuno-polymerase chain reaction detection. The data presented here shows a new systematic procedure that enables the fast generation of several competing haptens for the rapid development of sensitive heterologous immunoassays. PMID- 18409636 TI - Effects of child care center ventilation strategies on volatile organic compounds of indoor and outdoor origins. AB - Preschool children can be at risk from harmful volatile organic compounds (VOCs) exposures in child care centers (CCCs). However, the effectiveness of ventilation for mitigating indoor VOCs exposures from myriad sources in CCCs is unclear. We investigated relationships between indoor exposures and risks of VOCs from indoor and outdoor sources under differentventilation strategies in 104 tropical CCCs. Factor analyses identified five dominant source groups of which four were associated with indoor sources, and one was associated with both indoor and outdoor sources. Indoor VOCs exposures and risks associated with indoor sources were lower in naturally (NV) and hybrid ventilated (HB) CCCs compared to air conditioned CCCs (ACMV and AC). This is attributed to enhanced dilution via higher ventilation in NV and HB CCCs compared to ACMV and AC CCCs. Conversely, there were no discernible differences in VOCs exposures and risk associated with both indoorand outdoor sources across differentventilation strategies. The observations made in this study have implications of ventilation strategies used in other settings. To mitigate VOCs exposures and risk, it is important to identify their major indoor and outdoor sources first. PMID- 18409637 TI - Reversed-phase chromatography fractionation tailored to mass spectral characterization of humic substances. AB - Large-scale structural characterization of humic substances via mass spectrometry requires reduction of complexity within nominal mass and separation of isomers, i.e., prefractionation. Humic substances (here loosely defined to encompass all humic, humic-like, and humic-containing material) are notoriously difficult to fractionate. Equally challenging is deriving information on whether and howfractionation has occurred. Here, reversed-phase high-performance liquid chromatography was used to induce tailored fractionation of Suwannee River fulvic acid (SRFA) within nominal mass. The fractionation was optimized on synthetic standards thatdiffered in polarity and had elemental formulas similar to SRFA. Fractions were analyzed via electrospray ionization ion-cyclotron resonance mass spectrometry. Kendrick and Van Krevelen comparisons showed that fractionation occurred as predicted based on known molecular formula patterns. PMID- 18409638 TI - Extraction and optical fluorescence method for the measurement of trace beryllium in soils. AB - Beryllium metal and beryllium oxide are important industrial materials used in a variety of applications in the electronics, nuclear energy, and aerospace industries. These materials are highly toxic, they must be disposed of with care, and exposed workers need to be protected. Recently, a new analytical method was developed that uses dilute ammonium bifluoride for extraction of beryllium and a high quantum yield optical fluorescence reagent to determine trace amounts of beryllium in airborne and surface samples. The sample preparation and analysis procedure was published by both ASTM International and the National Institute for Occupational Safety and Health (NIOSH). The main advantages of this method are its sensitivity, simplicity, use of lower toxicity materials, and low capital costs. Use of the technique for analyzing soils has been initiated to help meet a need at several of the U.S. Department of Energy legacy sites. So far this work has mainly concentrated on developing a dissolution protocol for effectively extracting beryllium from a variety of soils and sediments so that these can be analyzed by optical fluorescence. Certified reference materials (CRM) of crushed rock and soils were analyzed for beryllium content using fluorescence, and results agree quantitatively with reference values. PMID- 18409639 TI - Laboratory tests on mercury emission monitoring with resonating gold-coated silicon cantilevers. AB - To measure extremely low concentrations of mercury vapor in gases as encountered in flue gases of coal-fired power plants, accurate and reliable online and/or portable mercury detection systems are needed. As discussed in this communication, resonating silicon-based cantilevers coated with thin films of gold change their resonant frequency when exposed to mercury vapors and could serve as the basis for such sensing devices. Two different types of commercial AFM cantilevers, which differed by physical dimensions and surface finish, were coated with a 10 nm film of gold and were tested in streams of argon containing mercury. The argon flow rates ranged from 5.7 to 57.4 ml/min, carrying mercury vapors at concentrations between 37 and 700 microg/m3. The results show that smaller cantilevers (approximately 140 microm x 40 microm x 4 microm) with a resonant frequency of 270-275 kHz were sensitive to less than 10 picograms of mercury, whereas larger cantilevers (approximately 245 microm x 50 microm x 7 microm) with a resonant frequency of 155-165 kHz have a sensitivity about 10 times lower. The results indicate that the kinetics of mercury capture by the gold coating follows a simple power law-correlation with the mass change (delta m) being proportional to t(n), where t is the capture time and n depends strongly on the concentration of mercury in the gas. It is also demonstrated that the mercury can be stripped off the gold coating by heating to 350 degrees C, which would allowthe cantilevers to be regenerated and reused. PMID- 18409640 TI - Plant growth-promoting bacteria for phytostabilization of mine tailings. AB - Eolian dispersion of mine tailings in arid and semiarid environments is an emerging global issue for which economical remediation alternatives are needed. Phytostabilization, the revegetation of these sites with native plants, is one such alternative. Revegetation often requires the addition of bulky amendments such as compost which greatly increases cost. We report the use of plant growth promoting bacteria (PGPB) to enhance the revegetation of mine tailings and minimize the need for compost amendment. Twenty promising PGPB isolates were used as seed inoculants in a series of greenhouse studies to examine revegetation of an extremely acidic, high metal contenttailings sample previously shown to require 15% compost amendment for normal plant growth. Several isolates significantly enhanced growth of two native species, quailbush and buffalo grass, in tailings. In this study, PGPB/compost outcomes were plant specific; for quailbush, PGPB were most effective in combination with 10% compost addition while for buffalo grass, PGPB enhanced growth in the complete absence of compost. Results indicate that selected PGPB can improve plant establishment and reduce the need for compost amendment. Further, PGPB activities necessary for aiding plant growth in mine tailings likely include tolerance to acidic pH and metals. PMID- 18409641 TI - Photocatalytic degradation of RhB by fluorinated Bi2WO6 and distributions of the intermediate products. AB - Fluorinated Bi2WOs catalyst was synthesized by a simple hydrothermal process. The effects of fluorine doping on crystal structure, optical property, photoinduced hydrophilicity, surface acidity, and photocatalytic activity of the as-prepared sample were observed in detail. Fluorinated Bi2WOs presented the enhanced photoactivity for the RhB degradation under the simulative sunlight (lamda > 290 nm), which could be a synergetic effect of the surface fluorination and the doping of crystal lattice. To get a better handle on the mechanistic details of this photocatalytic system, the photodegradation process of RhB was examined. In the fluorinated Bi2WO6 system, five intermediates, namely, N,N-diethyl-N' ethylrhodamine, N,N-diethylrohodamine, N-ethyl-N'-ethylrhodamine, N ethylrhodamine, and rhodamine were thus identified, whereas the first three intermediates could only be identified in the case of the Bi2WO6 system. This result indicated that more RhB molecules were degraded via the deethylation process in the fluorinated Bi2WO6 system. It was proposed that the (F-) containing function on the catalyst surface could serve as an electron-trapping site and enhance interfacial electron-transfer rates by tightly holding trapped electrons. On the basis of the experimental results, a photocatalytic mechanism was discussed in detail. PMID- 18409642 TI - Chromium(VI) reduction kinetics by zero-valent iron in moderately hard water with humic acid: iron dissolution and humic acid adsorption. AB - In zerovalent iron treatment systems, the presence of multiple solution components may impose combined effects that differ from corresponding individual effects. The copresence of humic acid and hardness (Ca2+/Mg2+) was found to influence Cr(VI) reduction by Feo and iron dissolution in a way different from their respective presence in batch kinetics experiments with synthetic groundwater at initial pH 6 and 9.5. Cr(VI) reduction rate constants (k(obs)) were slightly inhibited by humic acid adsorption on iron filings (decreases of 7 9% and 10-12% in the presence of humic acid alone and together with hardness, respectively). The total amount of dissolved Fe steadily increased to 25 mg L(-1) in the presence of humic acid alone because the formation of soluble Fe-humate complexes appeared to suppress iron precipitation. Substantial amounts of soluble and colloidal Fe-humate complexes in groundwater may arouse aesthetic and safety concerns in groundwater use. In contrast, the coexistence of humic acid and Ca2+/Mg2+ significantly promoted aggregation of humic acid and metal hydrolyzed species, as indicated by XPS and TEM analyses, which remained nondissolved (>0.45 microm) in solution. These metal-humate aggregates may impose long-term impacts on PRBs in subsurface settings. PMID- 18409643 TI - Membrane-aerated biofilm reactor for the treatment of acetonitrile wastewater. AB - A membrane-aerated biofilm reactor (MABR) was studied for the treatment of wastewater containing acetonitrile, a typical organonitrile compound. The MABR used hydrophobic hollow fiber membranes as the diffusers for bubbleless aeration as well as the carriers for biofilm growth. The objectives were to prevent the stripping-loss of acetonitrile during aeration and to achieve acetonitrile biodegradation plus nitrogen removal simultaneously in a single biolfilm on the membranes. In the MABR, oxygen and substrates were supplied to the biofilm from opposite sides, in contrast to those from the same side in conventional biofilm bioreactors. Operational factors, including surface loading rate and upflow fluid velocity in the bioreactor, on the effect of acetonitrile biodegradation performance were examined. The profiles of dissolved oxygen concentration and microbial activities and populations in the biofilm were investigated. Experimental results showed that, with the adapted microorganisms, removal of acetonitrile at approximately 98.6 and 83.3%, in terms of total organic carbon and total nitrogen, were achieved at a surface loading rate (in terms of membrane surface) of up to 11.29 g acetonitrile/ m2 x d with an upflow fluid velocity of 12 cm/s and a hydraulic retention time of 30 h. The biofilm on the membranes developed an average thickness of about 1.6 mm in the steady state and consisted of oxic/anoxic/anaerobic zones that provided different functions for acetonitrile degradation, nitrification, and denitrification. The acetonitrile-degrading bacteria in the MABR appeared to secrete more extracellular polymeric substances that enhanced the attachment and development of the biofilm on the membranes. The study demonstrated the potential of using the MABR for the treatment of organonitrile wastewater. PMID- 18409644 TI - Electroosmotic flow stimulates the release of alginate-bound phenanthrene. AB - There is growing interest in employing electro-bioremediation, a hybrid technology of bioremediation and electrokinetics for the treatment of contaminated soil. Most present applications of electrokinetics aim at pollutant extraction, which requires transport over large distances facilitated by electroosmotic flow (EOF). They do not explicitly account for the possibility that EOF passing along soil particles stimulates the release of hydrophobic organic compounds (HOC) and locally improves pollutant bioavailability. Here, we report on the stimulated release of polycyclic aromatic hydrocarbon (phenanthrene) from model organic matter in the presence of direct current (DC) electric fields (0.5-2 V cm(-1)) typically used in electrobioremediation measures. Alginate beads were employed as a model polymer release system (MPRS) exhibiting similar release behavior as natural organic matter (NOM). In the presence of EOF the phenanthrene release flux from alginate beads was between 1.4 and 1.8-fold higher than under hydraulic flow conditions with equal bulk water velocity and 30-120-fold higherthan under stagnantwater conditions. Our data suggest that DC-electric fields (0.5-2 V cm(-1)) can stimulate the release of PAH bound to particles exposed to stagnant water zones often found at hydraulic flow regimes restricted by low permeability. PMID- 18409645 TI - Reduction of dioxin-like compound emissions from a Waelz plant with adsorbent injection and a dual baghouse filter system. AB - Previous study indicates that the polychlorinated dibenzo-p-dioxin and dibenzofurans (PCDD/F) concentration measured in the stack gas of the Waelz plant investigated reached 194 ng-TEQ/(N m3) (TEQ = toxic equivalence), due to the relatively high potential of PCDD/F formation and a low PCDD/F removal efficiency (<70%) achieved with the baghouse filter (BF). In September 2006, the Taiwan government setthe PCDD/F emission limit for existing Waelz plants as 1.0 ng-I TEQO/(N m3). The retrofit technology for reducing PCDD/F emissions from the existing Waelz plant was evaluated at the same time. Carbon-type adsorbent injection technology was adopted in early 2006 to reduce the emission of dioxin like compounds at the Waelz plant investigated. Flue gases and ambient air samplings were conducted during the two stages of retrofit to evaluate the removal efficiency of dioxin-like compounds at the Waelz plant investigated. At stage 1, by applying adsorbent injection + single baghouse filter (SBF), the PCDD/F and polychlorinated biphenyl (PCB) concentrations measured in the stack gas at the Waelz plant were 4.62 ng-TEQ/(N m3) and 0.08 ng-TEQ(WHO)/(N m3) (TEQ(WHO) = World Health Organization TEQ), respectively, as the adsorbent injection rate was controlled at 40 kg/h (or 540 mg/ (N m3)). At stage 2, the PCDD/F and PCB concentration measured at stack gas, achieved with adsorbent injection + dual baghouse filter (DBF) system, were further reduced to 0.235 + 0.04 ng-I-TEQ/(N m3) (I-TEQ = International TEQ) and 0.004 + 0.002 ng-TEQ(WHO)/(N m3) with the adsorbent injection rate at 16 kg/h (or 215 mg/(N m3)). In the meantime, the atmospheric PCDD/F concentrations measured in the vicinity area of the Waelz plant were greatly reduced from 568-1465 to 48.9-130 fg-I-TEQ/m3. Higher removal efficiency (>99.8%) achieved at a lower adsorbent injection rate (16 kg/h) of the adsorbent injection + DBF system also significantly reduced the total PCDD/-F and PCB emission flows (per kg of electric arc furnace dust treated) to 1925 ng-I-TEQ and 30.5 ng-TEQ(WHO), respectively. PMID- 18409646 TI - Abiotic degradation of hexahydro-l,3,5-trinitro-1,3,5-triazine in the presence of hydrogen sulfide and black carbon. AB - We report that hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) was rapidly destroyed by sulfides in the presence of black carbon, forming nitrite and formaldehyde, rather than toxic nitrosated reduction products. Although traditionally viewed as inactive sorbents, black carbons have been noted to participate in the destruction of certain contaminants, such as azo dyes, via quinonoid groups. However, in our experiments sulfide modification of quinones did not seem to be involved. Although at least 1.2 mM sulfides were needed for the reaction to proceed, abiotic natural attenuation of RDX in marine sediments may occur, because these concentrations are found in certain marine sediments, together with black carbon. In the absence of natural black carbons, synthetic black carbons, such as activated carbon, may be added to sediments. As compared with other in situ techniques, such as bioremediation and zero-valent iron cutoff trenches, which often generate nitrosated byproducts, this in situ, abiotic technique may be an attractive alternative. PMID- 18409647 TI - Removal efficiency and binding mechanisms of copper and copper-EDTA complexes using polyethyleneimine. AB - Copper is used extensively in semiconductor circuits as the multilayer metal. In addition to copper, waste streams often contain chelating agents like EDTA, which is widely used in the process to enhance solubility of copper, and it tends to form copper-chelated complexes. PEI--agarose adsorbents in a packed-bed column are capable of removing these anionic complexes, but the competitive binding between this chelating agent and PEI for copper is not well understood and needs to be explored. The current work focuses on investigating copper sorption by PEI agarose adsorbent in the presence of EDTA. The pH of the column is fixed at 5.5 using 0.1 M acetate buffer. The ratio of chelator to copper ions is varied. Copper binding capacity and copper breakthrough curves are compared and contrasted to results without additional chelator present. An excess of EDTA leads to an increase in the fraction of free dissociated (anionic) ligand that competes for electrostatic attraction on protonated amine groups and therefore leads to a decrease in sorption capacity in the column. However, this waste treatment technique is still feasible for the semiconductor industry as large volumes of copper-contaminated solutions from actual waste can be concentrated 12 fold. When equimolar (copper to EDTA) or higher concentrations of EDTA are present, acetate can be utilized to recover the metal; for low ratios of copper to EDTA, metal recovery is achieved using hydrochloric acid. PMID- 18409648 TI - A new route for degradation of volatile organic compounds under visible light: using the bifunctional photocatalyst Pt/TiO2-xNx in H2-O2 atmosphere. AB - The bifunctional photocatalyst Pt/TiO2-xNx has been successfully prepared by wet impregnation. The properties of Pt/ TiO2-xNx have been investigated by diffuse reflectance spectra, X-ray diffraction, transmission electron microscopy, X-ray photoelectron spectroscopy, a photoluminescence technique with terephthalic acid, and electric field induced surface photovoltage spectra. The photocatalytic activity of the sample was evaluated by the decomposition of volatile organic pollutants (VOCs) in a H2-O2 atmosphere under visible light irradiation. The results demonstrated that nitrogen-doped and platinum-modified TiO2 in a H2-O2 atmosphere could enormously increase the quantum efficiency of the photocatalytic system with excellent photocatalytic activity and high catalytic stability. The increased quantum efficiency can be explained by enhanced separation efficiency of photogenerated electron-hole pairs, higher interface electron transfer rate, and an increased number of surface hydroxyl radicals in the photocatalytic process. A mechanism was proposed to elucidate the degradation of VOCs over PtTiO(2-x)Nx in a H2-O2 atmosphere under visible light irradiation. PMID- 18409649 TI - Removal of methyl parathion from artificial off-gas using a bioreactor containing a constructed microbial consortium. AB - Methyl parathion (MP), a highly toxic organophosphorus pesticide, was widely used for agriculture crop protection. During the production of MP and the process of MP-containing wastewater treatment, MP can release into the atmosphere and will do great harm to adjacent communities. A consortium comprised of an engineered microorganism and a natural p-nitrophenol (PNP) degrader was assembled for complete mineralization of MP. We genetically engineered Escherichia coli BL21 (DE3) enabling the overexpression of methyl parathion hydrolase (MPH). In addition, we isolated Ochrobactrum sp. strain LL-1 that utilized PNP, a product of MP hydrolysis, as the sole carbon, nitrogen, and energy source. The coculture effectively hydrolyzed 0.2 mM MP and prevented the accumulation of PNP in suspended culture. A laboratory-scale bioreactor containing the dual-species consortium was developed for the treatment of artificial off-gas containing MP. The bioreactor maintained over 98% of average MP removal efficiency over a 75 day period, and PNP produced from hydrolysis of MP was degraded completely, indicating that complete mineralization of MP was achieved. The strategy of linking degrading consortium to a bioreactor may provide an alternative to physicochemical abatement technologies for the treatment of waste-gas streams containing MP as well as other PNP-substituted organophosphates. PMID- 18409650 TI - Energy, greenhouse gas, and cost reductions for municipal recycling systems. AB - Curbside recycling programs can be more cost-effective than landfilling and lead to environmental benefits from the recovery of materials. Significant reductions in energy and emissions are derived from the decrease of energy-intensive production with virgin materials. In many cities, competing priorities can lead to limited consideration given to system optimal collection and processing strategies that can drive down costs and increase revenue while simultaneously reducing system energy consumption and greenhouse gas (GHG) emissions. We evaluate three alterations to a hypothetical California city's recycling network to discern the conditions under which the changes.constitute system improvements to cost, energy, and emissions. The system initially operates with a collection zoning scheme that does not mitigate the impact of seasonal variations in consumer tonnage. In addition, two collection organizations operate redundantly, collecting recyclables from different customer types on the same street network. Finally, the system is dual stream, meaning recyclables are separated at the curbside. In some scenarios, this practice can limit the consumer participation rate leading to lower collection quantities. First, we evaluate a "business as usual" (BAU) scenario and find that the system operates at a $1.7 M/yr loss but still avoids a net 18.7 GJ and 1700 kg of greenhouse gas equivalent (GGE) per ton of material recycled. Second, we apply an alternative zoning scheme for collection that creates a uniform daily pickup demand throughout the year reducing costs by $0.2 M/yr, energy by 30 MJ/ton, and GHG emissions by 2 kg GGE/ton. Next, the two collection organizations are consolidated into a single entity further reducing vehicle fleet size and weekly vehicle miles traveled resulting in savings from BAU of $0.3 M/yr, 100 MJ/ton, and 8 kg GGE/ton. Lastly, we evaluate a switch to a single-stream system (where recyclables are commingled). We showthat single-stream recycling can increase the total amount of material collected to a degree that lowers overall net cost ($0.2 M/yr) and leads to further reductions in energy use (210 MJ/ton) and emissions(16 kg GGEton). However,there can be circumstances in which maintaining a consolidated dual stream system is preferred over single stream. A sensitivity analysis is also performed and a discussion is presented addressing the applicability of this city network to others. PMID- 18409651 TI - Supercritical CO2 desorption of activated carbon loaded with 2,2,3,3-tetrafluoro 1-propanol in a rotating packed bed. AB - Desorption of activated carbon loaded with 2,2,3,3-tetrafluoro-1-propanol (TFP) by supercritical carbon dioxide in a rotating packed bed was investigated in this study. The experimental data show that the time required to achieve complete desorption of TFP from activated carbon in a rotating packed bed was much lower than that in a static packed bed. The reduction of desorption time is attributed to the presence of centrifugal force. The supercritical CO2 desorption efficiency in a rotating packed bed was observed to increase with increasing rotation speed, pressure, and C02 flow rate. To enhance desorption efficiency, a smaller activated carbon particle size was suggested. At low operating pressures such as 8.96 and 11.72 MPa, a better desorption efficiency was found to occur at lower temperatures in a temperature range of 305-335 K. However, at high operating pressures such as 15.86 MPa, a temperature of 315 K was found to be more appropriate for desorption, as compared to other temperatures. Due to a reduction of packed bed volume and an increase in desorption efficiency, supercritical CO2 desorption in a rotating packed bed is suggested for recovering TFP from the exhaust gases. PMID- 18409652 TI - Green roof valuation: a probabilistic economic analysis of environmental benefits. AB - Green (vegetated) roofs have gained global acceptance as a technologythat has the potential to help mitigate the multifaceted, complex environmental problems of urban centers. While policies that encourage green roofs exist atthe local and regional level, installation costs remain at a premium and deter investment in this technology. The objective of this paper is to quantitatively integrate the range of stormwater, energy, and air pollution benefits of green roofs into an economic model that captures the building-specific scale. Currently, green roofs are primarily valued on increased roof longevity, reduced stormwater runoff, and decreased building energy consumption. Proper valuation of these benefits can reduce the present value of a green roof if investors look beyond the upfront capital costs. Net present value (NPV) analysis comparing a conventional roof system to an extensive green roof system demonstrates that at the end of the green roof lifetime the NPV for the green roof is between 20.3 and 25.2% less than the NPV for the conventional roof over 40 years. The additional upfront investment is recovered at the time when a conventional roof would be replaced. Increasing evidence suggests that green roofs may play a significant role in urban air quality improvement For example, uptake of N0x is estimated to range from $1683 to $6383 per metric ton of NOx reduction. These benefits were included in this study, and results translate to an annual benefit of $895-3392 for a 2000 square meter vegetated roof. Improved air quality leads to a mean NPV for the green roof that is 24.5-40.2% less than the mean conventional roof NPV. Through innovative policies, the inclusion of air pollution mitigation and the reduction of municipal stormwater infrastructure costs in economic valuation of environmental benefits of green roofs can reduce the cost gap that currently hinders U.S. investment in green roof technology. PMID- 18409653 TI - Phylogeny and growth strategy as predictors of differences in cobalt concentrations between plant species. AB - Analyses reported here quantify the contribution of plant phylogeny and plant growth strategy to soil-to-plant transfer of Co. Estimated relative mean (ERM) Co concentrations in shoots of 241 species of flowering plant were derived using a residual maximum likelihood (REML) analysis. There were significant differences in, and a loge-normal frequency distribution of, ERM Co concentrations between species. A significant percentage of interspecies variance could be assigned to taxonomic categories above the species, (Family and above 21.5%; Order and above 12.22%). Time-series analysis of ERM Co concentrations ordered in the species sequence of the Angiosperm Phylogeny Group (APG II (2003)) revealed significant autocorrelation with an increase from Commelinid Monocot to Asterid Eudicot and a pronounced peak in the Core Eudicots. ERM Co concentrations categorized by plant growth strategy sensu Grime (2001) showed an increase toward stress-tolerant strategies. Plant species are not, therefore, independent units of Co concentration--factors derived from higher levels of biological organization exert significant effects. These effects can provide the basis of new techniques for selecting plant species for biotechnologies and for predicting the exposure of organisms to Co. They show that plant phylogeny and growth strategy might help refine predictions of soil-to-plant transfer of a variety of pollutants, and suggest research that might link molecular and higher level processes in contaminated soil-plant systems. PMID- 18409654 TI - Emissions from photovoltaic life cycles. AB - Photovoltaic (PV) technologies have shown remarkable progress recently in terms of annual production capacity and life cycle environmental performances, which necessitate timely updates of environmental indicators. Based on PV production data of 2004-2006, this study presents the life-cycle greenhouse gas emissions, criteria pollutant emissions, and heavy metal emissions from four types of major commercial PV systems: multicrystalline silicon, monocrystalline silicon, ribbon silicon, and thin-film cadmium telluride. Life-cycle emissions were determined by employing average electricity mixtures in Europe and the United States during the materials and module production for each PV system. Among the current vintage of PV technologies, thin-film cadmium telluride (CdTe) PV emits the least amount of harmful air emissions as it requires the least amount of energy during the module production. However, the differences in the emissions between different PV technologies are very small in comparison to the emissions from conventional energy technologies that PV could displace. As a part of prospective analysis, the effect of PV breeder was investigated. Overall, all PV technologies generate far less life-cycle air emissions per GWh than conventional fossil-fuel-based electricity generation technologies. At least 89% of air emissions associated with electricity generation could be prevented if electricity from photovoltaics displaces electricity from the grid. PMID- 18409655 TI - Investigation of mechanisms of polycyclic aromatic hydrocarbons (PAHs) initiated from the thermal degradation of styrene butadiene rubber (SBR) in N2 atmosphere. AB - This study has been carried out to characterize the thermal decomposition of styrene-butadiene rubber (SBR), using thermogravimetric analysis (TGA) coupled to online GC/MS, and to investigate the formation and ultimate fate of chemical species produced during gasification of SBR. A preliminary mechanistic understanding has been developed to explain the formation and relationship of light hydrocarbons (C1-C4), substituted aromatics, and polycyclic aromatic hydrocarbons (PAHs) during the decomposition of SBR in a N2 atmosphere. Identification and absolute concentrations of over 50 major and minor species (from hydrogen to benzo[ghi]perylene) have been established, and the measurements have been carried out between 300 and 500 at 10 degrees C/min heating rate in a N2 atmosphere. The concentration of styrene reached 120 PPMV and the concentration of other substituted aromatics, such as toluene and ethyl benzene reached 20 and 5 PPMV, respectively. These measurements indicate PAH formation at a relatively lower temperature as compared to conventional fuel, such as coal and diesel. The PAH sequence is not simply the constructing of larger PAHs from smaller ones to achieve the complex polymer structures. It is possible to generate large PAH molecules while circumventing the typical construction pathway. PMID- 18409656 TI - Linking molecular and population stress responses in Daphnia magna exposed to cadmium. AB - DNA microarrays can be used to measure environmental stress responses. If they are to be predictive of environmental impact, we need to determine if altered gene expression translates into negative impacts on individuals and populations. A large cDNA microarray (14000 spots) was created to measure molecular stress responses to cadmium in Daphnia magna,the mostwidely used aquatic indicator species, and relate responses to population growth rate (pgr). We used the array to detect differences in the transcription of genes in juvenile D. magna (24 h old) after 24 h exposure to a control and three cadmium concentrations (6, 20, and 37 microg Cd2+ L(-1)). Stress responses at the population level were estimated following a further 8 days exposure. Pgr was approximately linear negative with increasing cadmium concentration over this range. The microarray profile of gene expression in response to acute cadmium exposure begins to provide an overview of the molecular responses of D. magna, especially in relation to growth and development. Of the responding genes, 29% were involved with metabolism including carbohydrate, fat and peptide metabolism, and energy production, 31% were involved with transcription/translation, while 40% of responding genes were associated with cellular processes like growth and moulting, ion transport, and general stress responses (which included oxidative stress). Our production and application of a large Daphnia magna microarray has shown that measured gene responses can be logically linked to the impact of a toxicant such as cadmium on somatic growth and development, and consequently pgr. PMID- 18409657 TI - Avoidance of aluminum toxicity in freshwater snails involves intracellular silicon-aluminum biointeraction. AB - Silicon (Si) ameliorates aluminum (Al) toxicity to a range of organisms, but in almost all cases this is due to ex vivo Si-Al interactions forming inert hydroxyaluminosilicates (HAS). We hypothesized a Si-specific intracellular mechanism for Al detoxification in aquatic snails, involving regulation of orthosilicic acid [Si(OH)4]. However, the possibility of ex vivo formation and uptake of soluble HAS could not be ruled out Here we provide unequivocal evidence for Si-Al interaction in vivo, including their intracellular colocalization. In snails preloaded with Si(0H)4, behavioral toxicity in response to subsequent exposure to Al was abolished. Similarly, recovery from Al-induced toxicity was faster when Si(OH)4 was provided, together with rapid loss of Al from the major detoxificatory organ (digestive gland). Temporal separation of Al and Si exposure excluded the possibility of their interaction ex vivo. Elemental mapping using analytical transmission electron microscopy revealed nanometre-scale colocalization of Si and Al within excretory granules in the digestive gland, consistent with recruitment of Si(OH)4, followed by high-affinity Al binding to form particles similarto allophane, an amorphous HAS. Given the environmental abundance of both elements, we anticipate this to be a widespread phenomenon, providing a cellular defense against the profoundly toxic Al(III) ion. PMID- 18409658 TI - Comment on "Chronic disease and early exposure to air-borne mixtures. 2. Exposure assessment". PMID- 18409659 TI - Emerging role of HIFU as a noninvasive ablative method to treat localized prostate cancer. AB - The use of high-intensity focused ultrasound (HIFU) as a method for ablation of a localized tumor growth is not new. Several attempts have been made to apply the principles of HIFU to the treatment of pelvic, brain, and gastrointestinal tumors. However, only in the past decade has our understanding of the basic principles of HIFU allowed us to further exploit its application as a radical and truly noninvasive, intent-to-treat, ablative method for treating organ-confined prostate cancer. Prostate cancer remains an elusive disease, with many questions surrounding its natural history and the selection of appropriate patients for treatment yet to be answered. HIFU may play a crucial role in our search for an efficacious and safe primary treatment for localized prostate cancer. Its noninvasive and unlimited repeatability potential is appealing and unique; however, long-term results from controlled studies are needed before we embrace this new technology. Furthermore, a better understanding of HIFU's clinical limitations is vital before this treatment modality can be recommended to patients who are not involved in well-designed clinical studies. This review summarizes current knowledge about the basic principles of HIFU and its reported efficacy and morbidity in clinical series published since 2000. PMID- 18409660 TI - Biomarker CCSA-2 may provide accurate blood test for colorectal cancer. PMID- 18409661 TI - Management of pain in the older person with cancer. Part 2: treatment options. AB - Pain in older cancer patients is a common event, and many times it is undertreated. Barriers to cancer pain management in the elderly include concerns about the use of medications, the atypical manifestations of pain in the elderly, and side effects related to opioid and other analgesic drugs. The care of older cancer patients experiencing pain involves a comprehensive assessment, which includes evaluation for conditions that may exacerbate or be exacerbated by pain, affecting its expression, such as emotional and spiritual distress, disability, and comorbid conditions. It is important to use appropriate tools to evaluate pain and other symptoms that can be related to it. Pain in older cancer patients should be managed in an interdisciplinary environment using pharmacologic and nonpharmacologic interventions whose main goals are decreasing suffering and improving quality of life. In this two-part article, the authors present a review of the management of pain in older cancer patients, emphasizing the roles of adequate assessment and a multidisciplinary team approach. PMID- 18409662 TI - Systemic therapy for lung cancer brain metastases: a rationale for clinical trials. AB - Despite the high prevalence of brain metastases in patients with metastatic lung cancer, these patients have been excluded from enrollment in clinical trials of new therapeutic drugs. The reasons for exclusion have centered on concerns that the blood-brain barrier may impede drug delivery into brain metastases, that brain metastases confer a dismal survival for metastatic lung cancer patients, and that brain metastases carry risk for cerebrovascular hemorrhage. A focused, updated review of these issues, however, clearly shows that these particular concerns are unwarranted. An extensive review of clinical trials on the efficacy of chemotherapeutic agents against lung cancer brain metastases is also provided. This collective information describes an area in need of therapeutic development and supports an initiative to evaluate novel targeted therapies for lung cancer brain metastases. PMID- 18409663 TI - New therapeutic options in gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors have until recently had a uniformly poor prognosis with lack of effective drug therapies. These tumors usually have activating mutations in either KIT or PDGFR-alpha tyrosine kinase receptors. Over the past decade, imatinib (Gleevec), a selective tyrosine kinase inhibitor has become the standard of care for the first-line treatment of patients with unresectable and metastatic disease. For patients with imatinib-resistant disease or intolerant to the side effects of imatinib, sunitinib (Sutent), a multitargeted tyrosine kinase inhibitor was recently approved. For earlier-stage disease, status post-complete surgical excision, preliminary data seem encouraging for the role of adjuvant imatinib in prolonging patients' disease free interval. The impact of neoadjuvant drug therapy needs to be further classified and explored. With additional evaluation of other tyrosine kinase inhibitors and novel therapies against other molecular markers, the treatment paradigm for this malignancy should continue to evolve. PMID- 18409664 TI - [Health management of Saipem workers with projects involving abroad activities]. AB - In remote areas and in developing countries, where adequate health-care structures are few and sparse, Occupational Medicine contributes to guaranteeing workers' health. Companies like Saipem, involved in activities that are carried out in remote, inhospitable areas must ensure the safety and guarantee the health conditions of workers in relation to the risk factors connected with the job as well as with the environment in which it is performed. In such situations, Occupational Medicine addresses both the health aspects of the workplace and of the community, and is the pivot around which revolves the health-care support of workers employed abroad in the sense of protection and enhancement of health. The risks connected with work abroad are of three main types: 1) job-related risks; 2) risks connected with the environment; 3) risks related to the organization of work and the changes in the worker's daily life. The job-related risks are similar to those connected with analogous jobs performed elsewhere. The risks connected with the environment are related to adverse climatic conditions, extreme temperatures and unknown and often dangerous flora and fauna. The occupational physician is called upon to assess the suitability of workers for jobs that are based in remote areas. The main clinical conditions that can prevent issue of the Medical Fitness Certificate to workers for long-stay jobs abroad are discussed. PMID- 18409665 TI - [The evaluation of workplace visual characteristics and ophthalmologic requirements to define fitness to work]. AB - It's necessary to proceed to an evaluation of visual function to define principal ophthalmologic requirements and fitness to work in relation of workplace characteristics. We also have to consider the effects of age on visual function, the specific work risks on eyes and sight care especially for near sight. The evaluation of these parameters permits to verify the compatibility between the psychophysics characteristics of workers and the requests of tasks and then to define fitness to its work. Moreover to the visual functions we have to consider organic visual conditions like conjunctivitis, blepharitis, stenosis of lacrimal gland, dacriocystitis, retinic diseases, muscular motility alteration of the eyes, glaucoma. Our study shows the relation between ophthalmologic requirements of workers and some job groups like professional drivers, technical clerks, precision work, height level workers and proposes a standard method to define fitness to work. Our standard method permits a specific and homogeneous evaluation of fitness to work. PMID- 18409666 TI - [Evidence Based Occupational Medicine: ten year experience with audiometric examination in a handicraft company]. AB - This work describes the audiometry threshold assessment of 1000 workers employed in different artisan categories during a period of ten-year noise professional exposure. The hearing loss noise-induced rates were determined by analysing audiometric tests at the beginning of our period of study and after 5 and 10 years of noise exposure. Environmental noise exposures were on average 88 dB(A), but near 90 dB(A) in some work categories. Workers widely used hearing protection devices, nearly at 93%, during the period we studied. The Evidence Based Occupational Medicine should find out points of reference proving the efficiency and effectiveness of occupational physicians: in this case, a positive trend in the reduction of hearing loss rate will be expected to confirm the goodness of prevention practice. Our study suggests that the levels of protection so far accepted are not effective enough in order to reduce the incidence of noise induced hearing loss in the course of the years: in despite of most accredited predicting models for hearing conservation programs, a significant percentage of workers exposed to industrial noise continues to present a high incidence of hearing loss. The Evidence Based Occupational Medicine suggests that the proposed prevention activities carried out in the described factories were not enough effective. PMID- 18409667 TI - [Management of "complicated" work fitness judgements among health care workers]. AB - The occupational physician, performing health surveillance within a hospital, may face to some difficulties due to the variety and complexity of the tasks and the health risk factors of the health care workers. One of the hardest issue for occupational physician is to provide judgement on worker's fitness. Moreover, this task could be more complicated when a impaired worker could represent an hazard for his patients and colleagues. The authors will illustrate three critical clinical cases examined in Occupational Health Unit of Luigi Sacco Hospital, Milan; furthermore, the authors will show the difficulties and the applied solutions in order to provide the judgement on worker's fitness. PMID- 18409668 TI - [Freedom from smoking for health care workers. A project from Trieste]. AB - A tree years interventional study to modify smoking habits in health workers in Trieste province was planed in the collaboration of occupational health unit and Tobacco's Dependence Study Center. The aim of this paper is refer about preliminary data of the project started in 2007 regarding smoking habits in health workers of the Azienda per i Servizi Sanitari n.1 "Triestina" (ASS1) and the Azienda Ospedaliera Universitaria Ospedali Riuniti di Trieste (AOUTS). The project consist of several actions. i) information about risks and opportunities of project; ii) pursuance of the law 51 L 3/2003; iii) Occupational Health Unit and Tobacco's Dependence Study Center collaboration; iv) follow-up of the subjects that choose the disaccustom program. During occupational medical surveillance we collected the data related to 492 workers, 37% of the cases were smokers (180). The results of test of dependence to smoke (test di Fagestrom) showed an high dependence in 19% and an high motivation to stop smoke (test di Richmond) in 39% of the smokers. More than fifty percent of this subjects gave their adhesion to the disaccustom program. PMID- 18409670 TI - [Adequacy and reliability of orthoanalyzer Ergovision for job-fitness evaluation]. AB - The aim of this research was to evaluate the Ergovision Screener (ES) accuracy e validity by a confrontation with the conventional ophthalmic check (OC), for the medical evaluation of job fitness. A population of 100 VDU operators was considered. Each subject underwent randomly both the ES and the ophthalmic check visit. Several test carried out by the Ergovision Screener were not consistent with the conventional ophthalmic check. In a number of cases, high false positive ratio have been found, which could lead to unnecessary further examinations. For all these reasons we believe that the ES is not an appropriate instrument for the medical evaluation of job fitness. PMID- 18409669 TI - [Health surveillance on workers exposed to wood dust: usefulness of the assessment of the nasal mucosa]. AB - The purpose of this paper is to evaluate alterations in the nasal mucosa in workers that for professional purpose, are exposed, for a long period of time, to wood dust (WD). The increased frequency in alterations could underline a mechanism for chronic damage that could lead to cancer This study took into account 50 cabinet workers (EW) who had been exposed to WD for an average of 33 years and were compared to 48 controls (CC). A questionnaire regarding nasal symptoms was submitted, the nasal mucosa was examined by fibroscopy, secretions were valuated, cytogram from a nasal swap was also done. 44% of the EW and 33.4% of CC showed macroscopic alterations of the mucosa (PR 1,32 IC95% 0,79-2,19). The cytogram was altered in 24% of EW and in 12.5% of CC (PR 1,92 IC95% 0,78-4,71). In EW there was an abnormal significant increase in nasal secretions compared to CS, 28% vs 11,4% (PR 2,69 IC95% 1,05-6,89). The results do not confirm our hypothesis, but they show an unexpected prevalence of alteration in the CC. While waiting for further results, we express doubts in proposing routinary specialistic evaluation to all the EW to WD. At present it is hard to pin point indicators that could help reach an early diagnosis in the development of sinus nasal cancer. PMID- 18409671 TI - [The preventive ergophthalmic evaluation of work places in call centers: a useful instrument for the prevention of work related asthenopic symptoms]. AB - The study of a correct lay-out of work places equipped with VDT/PC in call centres, if carried out before the beginning of work activities needs an interdisciplinary cooperation among OH&S operators. However, such a preventive evaluation can relevantly reduce onset and severity of asthenopic symptoms frequently reported by this kind of operators. Consequently, such preventive evaluation can avoid the necessity of carrying out those complex and expensive structural lay-out modifications, which are frequently needed to reduce VDT/PC workers' asthenopic symptoms. PMID- 18409672 TI - [Analysis and evaluation of the visual effort in remote-control public traffic operators working with computer-based equipments]. AB - The aim of this study is the objective evaluation of the visual effort in 6 public traffic controllers (4 male, 2 female, mean age 29,6), by means of electronic equipment. The electronic equipment quantify the observation distance and the observation time for each controller's occupational visual field. The quantification of these parameters is obtained by the emission of ultrasound at 40 KHz from an emission sensor (placed by the VDT screen) and the ultrasound reception by means of a receiving sensor (placed on the operator's head). The travelling time of the ultrasound (US), as the air speed is known and costant (about 340 m/s), it is used to calculate the distance between the emitting and the receiving sensor. The results show that the visual acuity required is of average level, while accommodation's and convergence's effort vary from average to intense (depending on the visual characteristics of the operator considered), ranging from 26,41 and 43,92% of accommodation and convergence available in each operator. The time actually spent in "near observation within the c.v.p." (Tscr) was maintained in a range from 2h 54' and 4h 05'. PMID- 18409673 TI - [Work efficiency and lighting conditions: an experimental study]. AB - Thirty-two voluntary subjects were selected, not suffering either from any degenerative ophthalmic diseases or refraction and ocular motility alterations. Each subject underwent close visual task experimental sessions (e.g. PC usage), under monitored experimental conditions. Aim of the study is the assessment of working efficiency effects caused by lighting conditions characterized by "according to law" illuminations, yet in presence of high or low luminance ratios in the occupational visual field". An analysis of the data showed that high luminance ratios conditions show a decrease of the performance (decrease overall efficiency, increase in the number of errors and time of execution), which where not detected with low luminance ratios conditions. Asthenopia did not show clear differences, possibly due to the effects of the intense near work which was present in both the experimental sessions. PMID- 18409674 TI - [Extended high frequency audiometry in the prevention of noise-induced hearing loss]. AB - An early detection of noise induced hearing loss (NIHL) may allow more effective protection measures. Our aim was to investigate the usefulness of high-frequency audiometry to evaluate the possibility of a future use of the high frequencies audiometry as an early indicator for noise induced hearing loss. A cross sectional study was performed involving 204 industrial noise exposed and 100 non industrial noise-exposed workers. Each subject was tested with both conventional frequency (0.25-8 kHz) and high-frequency (9-18 kHz) audiometry during the annually health surveillance campaign conducted in two Italian cement factories. As expected, noise exposed workers were found to have significantly higher hearing thresholds (P < 0.05) at both conventional and extended high frequencies. Marked differences were found for EHFA. Moreover, significant differences at EHFA were detected also in the subgroup of noise-exposed workers with normal findings at conventional audiometry. Our finding indicate that the use of the extended high frequency test may represent a useful tool for detecting early changes of hearing impairment and that it could be used in addition to the conventional test to better prevent the progression of noise hearing loss. PMID- 18409675 TI - [Carbo-dehydrated transferrin (CDT) as a marker of alcohol intake: problems with analysis and interpretation. Case report]. PMID- 18409676 TI - [Evaluation of professional exposure to chloride vinyl monomer and vinyl idene chloride for a pharmaceutical packaging worker]. AB - The study was conducted by Judicial Policy investigations of Prosecution's Office. The event was connected by a professional founded suspicion disease of a pharmaceutical worker. First information coming from the Authority indicated a chloride vinyl monomer (CVM) exposure. We applied a chemical risk assessment method to estimate real professional exposure. The method was based on the productive cycle, physical and chemical and toxicological properties. The method combined to environmental data permitted to formulate etiological hypothesis. The worker during drugs packaging was exposed to CVM and vinylidene chloride (CVDM) caused by blister warming and by glue deposition. We explain the evaluations by which we could consider the pollutant different distribution in workplaces. PMID- 18409677 TI - [Italian Program for Surveillance of Acute Pesticide-Related Illnesses: cases identified in 2005]. AB - In 2005, the Italian System for Surveillance of Acute Pesticide-Related Illnesses (SIAF) identified 625 cases, among which 520 unintentionally exposed. The majority of these subjects were men (75%) and aged 26-65 years (65%). About 63% of all exposures occurred at work. Severity for these illnesses was low for 94% and moderate for 5%. Four cases were classified as illnesses of high severity. Some 70% of all the reported exposures occurred between May and September. The active ingredients responsible for the largest number of cases were: glyphosate (n. 56), copper sulphate (n. 55), methomyl (n. = 52), metam-sodium (n. 24). Three episodes of collective environmental exposure to soil fumigants involving 23 subjects were also detected. PMID- 18409678 TI - [Occupational exposure to formaldehyde in autopsy room and in pathologic anatomy laboratories]. AB - The aim of the present study is to evaluate the personal exposure to formaldehyde in an autopsy room and in three pathological anatomy laboratories. Passive sampling for the whole workshift and HPLC analysis were performed. The final result showed a remarkable exposure of the workers enrolled. All the data obtained exceeded the NIOSH-TWA and several cases personal of exposure levels above the ACGIH-Ceiling value (0,37 mg/m3) were observed in the anatomy laboratory. PMID- 18409679 TI - [Biological monitoring in workers exposed to inorganic arsenic in a disused industrial plant in the area of Manfredonia]. AB - Inorganic arsenic and its methylated metabolities were measured in 108 spot urine samples obtained from the medical surveillance programme of workers exposed to inorganic Arsenic in July 2006. 15% of the samples showed levels higher than limit value of 35 microg/L (mean value 23,9 microg/L). After the improvement of the working conditions, in August-October 2006, we collected a urinary sample from each of the 108 workers enrolled. A questionnaire was also administrated, in order to investigate the influence of occupational and non occupational factors on the urinary arsenic excretion. The median value of urinary arsenic was 15,12 microg/L; among the 108 samples, 5% showed levels higher than limit value. A significant difference was observed in relation with sea-food consumption and aging stratification. In conclusion, we have described a significant reduction of urinary arsenic excretion between the two phases of biological monitoring, likely due to a proper hygienic work-related intervention. PMID- 18409680 TI - [Exposure to ammonium persulphate by inhalation: effect on the NANC inhibitory neurotransmitters in the guinea pig trachea]. AB - To evaluate the effect of ammonium persulphate (AP) inhalation on NANC inhibitory (i-NANC) neurotransmitters of guinea pig airways, we exposed eight guinea pigs to AP (1 mg/m3), by aerosol inhalation for 30 minutes daily for three weeks. Control animals inhaled saline aerosol. After the last exposure, the isolated trachea was mounted in an organ bath and electrically stimulated in the presence of hyoscine, piperoxane and propranolol. The i-NANC responses were evaluated as decreases in intraluminal pressure and expressed as area under the curve (AUC, Pa x seconds). The isolated tracheae were treated with a-chymotrypsin, L-NAME, zinc protoporphyrin IX and ODQ, that inhibit the production or action of the single neurotransmitters, like peptides, NO and CO. In the exposed individuals, the NANC relaxations were below 50%, as compared to controls (P < 0.01). NO and CO were the neurotransmitters responsible for all the i-NANC responses, in similar proportions either in exposed individuals or in controls. In conclusion, ammonium persulphate exposure impairs the i-NANC control of airway tone without specifically affecting any neurotransmitter. PMID- 18409681 TI - [Modifications of hepatic transaminases in workers exposed to low doses of isopropanol]. AB - Isopropanol (IPA) is a volatile solvent that is used in many industrial process. The major symptoms of acute isopropanol toxicity include dizziness, incoordination, headache, hypothermia, eye ataxia, irritation of upper respiratory tract and shortness of breath. Vomiting, hematemesis, diarrhoea and hypotension may occur following accidental ingestion of IPA. No data regarding subchronic or chronic toxicity of IPA were identified. The aim of this study was to measure the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and of gamma-glutamyltransferase (gamma-GT) of the last five years in 40 printer workers after the removal of IPA from the industry. The serum levels of ALT, AST and gamma-GT were higher in the exposed workers than in non exposed. In conclusion, the results of this study show that the removal of IPA from the industry had a positive health effect improving the hepatic function of the workers. PMID- 18409682 TI - [Occupational exposure to manganese in ferroalloy industry: neurobehavioral effects in a workers' cohort]. AB - Our Institute has been following for 20 years a group of workers of a ferroalloy industry in order to evaluate neurobehavioral effects due to manganese exposure. Five years after the last study we have planned another one, to evaluate differences in neuromotor e cognitive functions between exposed and controls and to perform a longitudinal evaluation of the results. Environmental and biological sampling were collected, liver and kidney functionality, haemochrome, iron metabolism and sieric prolactine were evaluated. Several tests were administered: postural evaluation, tremor, four tests of the SPES battery, Pursuing Aiming, five tests of the Luria Nebraska Motor Battery, Raven Progressive Matrices, Trail Making Test, Mood Scale, Brief Symptoms Inventory, neuropsychological symptoms questionnaire. Personal habits and working, living and clinical histories were collected. We evaluated 43 exposed workers and 40 controls. Exposure indicators resulted all significantly higher in exposed workers. Neuropsychological examination showed differences in Raven Progressive Matrices and Pursuit Aiming, higher tremor values and differences in postural evaluation between exposed and controls. PMID- 18409683 TI - [Fibers glass induced cytotoxicity and genotoxicity]. AB - Man-made vitrous fibers, have been widely used as a substitute for asbestos, as an insulation material. However the fibrous morphology of MMVFs raises concern about potential health hazard. The aim of our study was to assess cytotoxic and genotoxic effects induced on a human alveolar cell line A549 by exposure to glass wool fibers (GW). Cells were exposed for 72 h to 5, 50, 100 microg/ml of glass wool, after incubation the cell viability was determined by a MTT reduction assay. The genotoxic effect was studies by Comet test. An undamaged cell appeared as a nucleoid and a cell with damaged DNA as a comet. Measurement of Comet parameters: % DNA in the tail, tail length and tail momente (the product of relative tail intensity and lenght, that provides a parameter of DNA damage) were obtained from the analysis. A MTT assay indicated that glass wool caused a decrease in cell viability and this decrease was concentration-dependent. The results of the Comet test for DNA damage detection indicated in cell exposed to glass wool fibers a significant increase of mean TM value. All these results provide that the glass wool fibers can induce cytotoxicity and genotoxicity PMID- 18409684 TI - [Occupational and environmental exposure to anilide and dicarboximide pesticides]. AB - Dicarboximide fungicides (DF) such as vinclozolin, iprodione, procymidone are widely used on vines, fruit and vegetables, and anilide herbicides (AH) such as diuron, linuron, propanil are used to control weeds on hard surfaces, such as, roads, railway tracks, paths, and in crops, forestry. Italian reports on food safety found many samples contaminated by pesticides belonging to these categories, even though only few exceeding L.M.R. Since adverse effects on human health, such as endocrine disruption, have been reported, biological monitoring is essential for exposure assessment both of occupationally exposed subjects and of the general population. Common metabolites of DF and AH are dichloroanilinines such as 3,4-DCA and 3,5-DCA, urine samples from 153 subjects living in Novafeltria, central Italy, were collected for analysis of 3,4- and 3,5-DCAs, each participant was invited to complete a very detailed questionnaire. A total of 151 out of 153 samples were found to be positive for 3,5-DCA, and 81.7% were positive for 3,4-DCA. Also 33 workers, engaged in application of propanil on rice in northern Italy, were involved in the study and 3,4-DCA was determined as marker of exposure. 3,4 and 3,5 dichloroaniline are useful and promising biological indicators for monitoring occupational and environmental exposure to these classes of pesticides. PMID- 18409685 TI - [Hearing function and solvent exposure: study of a worker population exposed to styrene]. AB - The aim of the study was to investigate the ototoxic effects of occupational styrene exposure, in absence of other risk factors. Pure-tone audiometric thresholds of 32 workers exposed to styrene, but not to noise, in fibreglass reinforced plastic boat manufacturing process were detected and compared to audiometric thresholds of a control unexposed group composed by 60 subjects. Exposure to styrene was measured by urinary mandelic + phenylglyoxylic acid (mean value 149 mg/g crea, SD 80 mg/g crea). For all the frequencies investigated (0,5 1-2-3-4-6-8 KHz) the exposed group showed slight higher mean (median) audiometric thresholds (p < 0.05) compared to controls matched by age and sex, except for 8 KHz in the right ear. The present experience seems to confirm the hypothesis that styrene exposure alone can determine a weak sensorineural high-frequency hearing loss. Such slight impairment, even if statistically significant, does not remarkably limit social hearing and do not involve legal medical aspects. Sample expansion and objective diagnostic tests (auditory brainstem evoked potentials, acoustic otoemissions) are needed. PMID- 18409686 TI - Neurological and neuropsychological features in Parkinsonian patients exposed to neurotoxic metals. AB - BACKGROUND: Environmental exposure to heavy metals and especially manganese (Mn) took place in Valcamonica, Italy, where a high prevalence of Parkinsonism was observed (age and sex standardized 407/100,000; 95% CI: 393.87-420.12), and the Standardized Morbidity Ratios was associated with environmental Mn levels. METHODS: A cross sectional study compared Parkinsonian patients residents in Valcamonica with patients from Brescia, Italy. Age- and sex-matched healthy individuals were recruited as controls. The protocol included information on clinical, occupational, residential history and life habits, neuro-psychological testing, and assessment of genetic polymorphism. RESULTS: The target group included 65 patients and 52 controls from Valcamonica, 28 patients and 14 controls from Brescia. Age at onset of the disease was lower in women from both areas. After adjusting for age and age at onset, patients from Valcamonica showed more severe motor impairment at the UPDRS scale, higher damage of cognitive and motor functions at MMSE, Token and Trial Making tests. Genetic variables showed a different allelic distribution of DRD4 gene between cases and controls, outside Valcamonica, where a less frequent familiarity for parkinsonism was reported. CONCLUSIONS: Parkinsonian patients with previous exposure to metals showed a more severe neuropsychological phenotype, without detectable contribution from genetic factors. PMID- 18409687 TI - [Exposure to polycyclic aromatic hydrocarbons (PAH) in rubber forming industry]. AB - Among various chemical agents present at the workplaces in the rubber industry, a particular attention was adressed to the Polycyclic Aromatic Hydrocarbons (PAH) contained in oil and carbon black, but some questions regarding level of exposure are also controversially discussed. The literature reports that PAH may have irritant effects; moreover some of these have been recognized as probably or possibly carcinogenic to human by the International Agency for Research on Cancer, the European Union, and other institutions. In Lombardy, a study aimed to evaluate the occupational exposure in Benzo[b]fluorantene the rubber forming industry was planned during last two years. The results of environmental air monitoring (the 16 most relevant, according to the American Environmental Protection Agency, EPA) and biological monitoring (urinary 1-hydroxypyrene excretion) show that PAH exposure in these workers is not higher than that observed in other study regarding low level and acceptable exposure. PMID- 18409688 TI - [Biological monitoring in the molding of plastics and rubbers]. AB - This survey was carried out in the molding of plastics and rubbers, in the "Professional Cancer Prevention Project" sponsored by the Lombardy region with the objective of developing and implementing protocols for evaluating exposure to carcinogens through the biological monitoring. The realities of molding the thermoplastic polymer ABS, rubber, and thermosetting plastics containing formaldehyde were examined. The carcinogenic substances identified in these processes were: 1,3-butadiene, acrylonitrile and styrene in molding ABS, polycyclic aromatic hydrocarbons (PAH) in molding rubber, and formaldehyde in molding the thermosetting plastics. Only for some of these substances biological indicators are available. The limited exposure to airborne chemicals in molding ABS and the intrinsic characteristics of biological indicators available for 1-3 butadiene have determined the non applicability of biological monitoring to this situation. The absence of a biological indicator of exposure to formaldehyde has made this situation not investigable. Exposure in the rubber molding was studied in 19 subjects applying the determination not metabolized PAH in urine. The levels of these indicators were similar to those measured in other groups of subjects without occupational exposure to PAH, confirming a low airborne contamination in this workplace. PMID- 18409689 TI - [In vitro study of genotoxic and oxidative effects induced on human pulmonary cells by exposure to PAHs extracted from airborne particulate matter collected in a coke plant]. AB - Genotoxic and oxidative effect of airborne particulate matter collected in a coke plant were evaluated on lung epithelial cells (A549). We aimed to clarify the mechanism of action of complex mixtures of PAHs and to identify biomarkers of effect of lung cancer. Particulate matter was analysed by GC/MS. Genotoxic and oxidative effects induced by the exposure to the extract were evaluated by Fpg comet assay. The cells were exposed for 30 min, 2h and 4h to 0.01%, 0.02% and 0.05% of the extract. We evaluated comet percentage and analysed tail moment values of exposed and unexposed cells treated with Fpg enzyme (TMenz) and untreated (TM) that indicate respectively oxidative and direct DNA damage. We found 0.328 ng/m3 of pyrene, 0.33 ng/m3 of benzo(a)anthracene, 1.073 ng/m3 of benzo(b)fluoranthene, 0.22 ng/m3 of benzo(k)fluoranthene, 0.35 ng/m3 of benzo(a)pyrene, 0.079 ng/m3 of dibenzo(a,h)anthracene and 0.40 ng/m3 of benzo(g,h,i)perylene. A dose-dependent increase, although not significant, of TM and TMenz in the exposed cells in respect to controls was found that indicates a slight increase of both direct and oxidative damage in exposed cells. A slight increase of comet percentage was found at the highest dose. We show the high sensibility of comet assay to measure early DNA damage also at low doses suggesting the use of such test on A549 to evaluate on target organ the effects of complex mixtures of genotoxic substances. PMID- 18409690 TI - [Significance and limitation of creatinine adjustment for urinary chromium and arsenic in biological monitoring of occupational exposure to these metallic elements]. AB - Creatinine adjustment has been used to remove the influence of the effect of urine dilution on exposure biomarkers measured in spot samples. This research aimed to determine the reliability of creatinine adjustment for urinary chromium and arsenic in subjects from general population considering interferences able to influence creatinine excretion. 444 male subjects were examined and each participant was administered an anamnestic questionnaire. Chromium and arsenic were determined on second morning void urine samples by atomic absorption spectrophotometry and creatinine by Jaffe method. Urinary creatinine showed a significant negative decrease with age increasing (p < 0.001). Chromium concentrations resulted higher in subjects with renal disease, particularly after creatinine adjustment (p = 0.014). Age was negatively correlated with urinary creatinine (rho = -0.23; p < 0.001), and positively with chromium (rho = 0.13; p = 0.007) and arsenic (rho = 0.17; p = 0.004), only after creatinine adjustment. A positive correlation was also found between unadjusted chromium and arsenic (rho = 0.10; p = 0.039) and between urinary creatinine and both unadjusted chromium (rho = 0.32; p < 0.001) and arsenic (rho = 0.18; p < 0.001). An high coefficient of correlation was observed between unadjusted chromium (rho = 0.88; p < 0.001) and arsenic (rho = 0.90; p < 0.001) and the respective adjusted values. Multiple regression showed a dependence of urinary creatinine and adjusted chromium and arsenic concentrations on age. In conclusion, although chromium and arsenic seem to have a renal kinetics of excretion comparable to the creatinine one, the influence of age on creatinine elimination suggests that the expression of urinary values of these metallic elements as microg/L is more reliable. PMID- 18409691 TI - [Influence of cigarette smoking on the excretion of urinary benzene in filling station attendants]. AB - The influence of cigarette smoking on concentrations of urinary benzene, a sensitive and specific biomarker proposed for biological monitoring of exposure to very low doses of benzene, was investigated in 24 filling-station attendants and 31 workers non occupationally exposed to benzene. Environmental monitoring was performed by personal passive samplers "Radiello", and a spot urine sample was collected at the end of the work shift, from all subjects, for the determination of urinary benzene. Exposure to benzene resulted significantly higher in filling-station attendants (mean 23.3 +/- 17.0 microg/m3; range 4.5 66.3 microg/m3) than in controls (mean 4.6 +/- 2.6 microg/m3; range < 3-11.5 microg/m3), while concentrations of urinary benzene did not show any significant difference between the two groups. Considering all subjects as a single group, it was observed that urinary benzene concentrations were positively correlated with the number of cigarettes smoked during the sampling time (rho = 0.38; p = 0.047) and with airborne benzene levels (rho = 0.32; p = 0.019), and negatively correlated with the time elapsed between the last smoked cigarette and urine collecting (rho = -0.40; p = 0.045). Multiple regression analysis confirmed the influence of cigarette smoking on urinary benzene concentrations. In conclusion, our study showed the validity of urinary benzene as a biomarker for biological monitoring of exposure to very low doses of benzene, although cigarette smoking determined a prevalent etiological role at the low environmental benzene concentrations observed. PMID- 18409692 TI - [Metals and oxidative stress in Parkinson's disease from industrial areas with exposition to environmental toxins or metal pollution]. AB - BACKGROUND: Parkinson's disease (PD) is characterized by a progressive degeneration of the nigrostriatal dopaminergic pathway resulting in movement disorders. PD is a complex disease, in which and environmental factors, as exposure to toxins or metals coul be involved. OBJECTIVE: To assess if serum metals (Cu, Fe, Zn), biological variables of their metabolism, total peroxides and antioxidants were abnormal in PD, in relation to environmental exposure. METHODS: We compared levels of serum copper, iron, zinc, ceruloplasmin and transferrin, peroxides, antioxidants (TRAP) in 65 PD patients coming from an Industrial zone highly exposed to metal pollution (Valcamonica) with measures from 28 PD patients from no metal pollution areas of the province of Brescia and 52 healthy controls coming from Valcamonica and 24 from the province of Brescia. RESULTS: PD patients had higher serum concentration of zinc than controls. Only in PD patients coming from Valcamonica levels of Cu were higher than in subjects coming from the province of Brescia. Moreover, In patients with PD levels of sieric Cu significantly correlated with score of the Unified Parkinson's Disease Rating Scale (UDPRS). CONCLUSIONS: Zinc seems to be higher in PD independently from the exposition to metal pollution. Perturbation of copper metabolism in PD seems to be related to exposition to environmental toxins or metal pollution and coul be involved in the progression of the disease itself. PMID- 18409693 TI - [Biomechanical overcharge of the upper limbs in hairdressers: from the task analysis to the job/exposition matrix]. AB - The analysis of professional diseases denounced from hairdressers in Italy and in Tuscany shows among these workers a reduction of some "typical" work related diseases, like dermatitis, asthma, etc. and a raise of upper limbs disorders. The upper limbs risk assessment process is very tough for the hairdresser's activity, because the working cycle includes different tasks (shampoo, cut, dyeing, etc.) and their combination in a working day is related to customers requests. The job illustrates the first results of a project started last year from the Tuscany Technical Advisory Department for Risk Assessment and. Prevention (CONTARP) of the Italian Workers' Compensation Authority (INAIL) and PISLL "G. Pieraccini" - ASL 10 of Florence. Through a study conducted with the OCRA Check List method on a sample constituted by 12 employees of five shops of the Florentine territory, we have arrived to a job-exposure matrix that allows to draw an index of exposure to the specific risk for every task, keeping in mind the contributions of the single repetitive assignment performed by the operator during the working day. PMID- 18409694 TI - [The adoption of electrical beds in hospital clinic units: evaluation of the impact on workers' low back pathologies]. AB - With the aim of reducing the number of occupational accidents and of judgements of limited work ability among the nurses having as work task the movement of not self-sufficient patients, the Legnano Hospital has defined a project of substitution of the traditional hospital beds with electrical beds. We describe the project of substitution of the beds and the positive results both on the health status of the workers using electrical beds and on the reduction of the falls occurred to the patients. PMID- 18409695 TI - [The posturological department as clinical support for occupational medicine: clinical cases and results of a hospital unit]. AB - The posturologic visit is a not widely known medical method for the evaluation and the therapy of low back pain. We describe the clinical and instrumental method followed in our posturological clinical unit organized jointly by hospital and university and the clinical cases evaluated in two years. An individual diagnostic evaluation and a personal therapy allowed an almost generalized complete remission of the symptoms and a very low number of reactivation of low back pain after a follow up of one years. PMID- 18409696 TI - [Back school project in a company: how to prevent low back pain]. AB - This study refers to the experience of Back School training which was performed inside the Fire Brigade Department of the multicompany Petrochemical Plant of Ferrara. Our project includes integrated activities that promote employee's well being related to their workplace and their work duties with the intention of improving how the organization runs. This project has been carried out in cooperation with the Reahabilitation Medicine Department "S. Giorgio" of Arcispedale S. Anna in Ferrara. PMID- 18409697 TI - [The posturological clinic evaluation in vdt/pc operators: a second level diagnostic instrument in low back pain evaluation]. AB - The diagnosis and the therapy of the cases of low back pain in absence of organic spinal column diseases, frequently referred by vdt/pc users in connection with their work activities are frequently unsatisfactory both for the Occupational Doctors and for the patients. The specialized posturological evaluation, composed by a specific clinical examination and by specific instrumental exams, seems to be a valid instrument for a more effective management of patients affected by such clinical cases. The treatments described in this research have eliminated or greatly reduced the pain in these patients, with an efficacy prolonged until at least a year; as it is demonstrated by the few relapses referred by the workers. We think that a more frequent cooperation between Occupational Doctors and Posturologists can improve both the clinical conditions of the patients and the possibilities they have in the years to maintain their complete work ability. PMID- 18409698 TI - [Carpal tunnel syndrome in handicraft planning]. AB - The purpose of this study is to assess the prevalence of carpal tunnel syndrome (CTS) in a group of bakers and to evaluate the presence of a biomechanical risk for upper limbs in the technological cycle. Health assessment (history, clinical examination, upper limbs electromyography) and risk evaluation through Check List OCRA (Occupational Repetitive Actions - Colombini / Occhipinti) have confirmed the initial hypothesis, placing this profession between those at risk for carpal tunnel syndrome. PMID- 18409699 TI - [Active search of work related tumours: preliminary findings]. AB - The OCCAM (Occupational Cancer Monitoring) project enabled the active detection of occupational cancer cases in Lombardy Region. METHODS: OCCAM is based on a record linkage with social security files to obtain occupational histories for all subjects having worked in private firms, since 1974. It provides risks by area, site and job. RESULTS: 271 incident cancer cases obtained by hospital discharge record in the period 2001-2002 where investigate to assess eventually their occupational origin. Approximately 38% where considered to be occupational cancers. CONCLUSIONS: OCCAM provides name of the firms and their economic activity completed by information coming from OCCAM risks ascertainment and deeper knowledge on productive cycle retained by local occupational health services. Thus this system can lead to detection of many cancer cases of occupational origin suitable for compensation and determine strategies for the improvement of the work environment. PMID- 18409700 TI - [Active search of work related diseases: the example of carpal tunnel syndrome in Pordenone's area]. AB - In the recent years working related pathologies of the upper limbs have raised at the top of ranking of diseases reported to INAIL in the Northern East Area of Italy. In 2004 658 cases of tendinitis have been filed and 361 cases of the Carpal Tunnel Syndrome (CTS) reported on a total of 1317 and 864 cases respectively in the whole Italian territory. The phenomenon (6.8% of total occupational disease in Pordenone province) interests essentially women and various working environments widely represented. We have worked together with an important Diagnose and Treatment Medical Centre to find new cases of professional illnesses, initially focusing our efforts on the CTS. We have identified 26 new cases of CTS as possibly originated from working conditions and we have filled 11 reports to the Judicial Authorities. In a training meeting with Orthopedic Doctors we have disclosed the results of our research while at the same time we have also divulged our knowledge on the importance of the working environment in the onset of the CTS providing the tools for a correct reporting of suspect cases. PMID- 18409701 TI - [Differences in the prevalence of work related disorders among construction workers between the end of 90s and 2006]. AB - The aim of this work is to analyse the prevalence of occupational diseases in construction industry, comparing the results of the first years of health surveillance (1996-2000) and today's situation. We compare the prevalence of occupational diseases observed in 1348 workers during the period 1996-2000 (mean age 36,4 years, DS 11,8; mean experience in construction industry 16,9 years, DS 12,3) and the one observed in 795 different workers during 2006 (mean age 38,8 years, DS 11,1; mean experience in construction industry 18 years, DS 16,6). The prevalence of occupational diseases is respectively 12.09% and 12.83%. But analysing the single pathology it's possible to observe important differences between the two groups. In the workers observed in 2006 there is a significant reduction in prevalence of hearing loss caused by chronic exposure to noise and in prevalence of vascular disease caused by hand-arm vibrations: respectively from 94,9 to 79,2 cases/1000 workers and from 6,7 to 2.6 cases/1000 workers. Musculoskeletal diseases instead are increased from 11,1 to 37,7 cases/1000 workers, showing more attention to the problem, better diagnostics ability and more recognition by public insurance agency for occupational diseases. PMID- 18409702 TI - [Epidemiologic surveillance in occupational bladder cancer: a Tuscan experience]. AB - The percentage of bladder cancer as occupational disease in West-Europe is of 5/10%, but only a few amount of them are recognized as occupational disease from INAIL. The above mentioned research project is realized in order to decrease the gap between expected and claimed cases of occupational disease and it is conducted with the collaboration of ASL of Pisa, ASL of Empoli, Azienda Ospedaliera Universitaria Pisana and INAIL. 677 patients with bladder cancer were interviewed by phone, among them 64 subjects had a working experience compatible with neoplastic risks because had a previous occupational exposure to aromatic amines and metal working fluids. These cases were discussed into a Medical Staff and 40 cases were considered "probable" for occupational disease, 18 "possible", 3 cases are suspended for more research, 3 cases are considered "no professional disease". The research allows finding out a great number of bladder cancer, increasing the total amount of workers with occupational disease. The integrated approach with the collaboration among different institutions is surely the best way to allow and guarantee a suitable and right protection of workers with occupational disease. PMID- 18409703 TI - [Transitional bladder cancer and occupational exposure. Accuracy assessment of a screening method based on structured interview]. AB - The transitional bladder cancer is the most frequent urinary malignant neoplasm. The main risk factors are cigarette smoking, recurrent inflammatory diseases of the urinary tract, assumption of some drugs, familiarity and occupational exposure to aromatic amines and polycyclic aromatic hydrocarbons. The aim of this study is to assess the screening efficacy of a short structured interview to detect the possible occupational bladder cancer, to submit to the specialists evaluation. The sample of the study is represented by 100 subjects, hospitalized for transitional bladder cancer in the years 2004 and 2005 at the Department of Urology of the Ospedale di Circolo - Fondazione Macchi in Varese. Through the structured interview was possible to estimate two occupational exposure indexes: Length-Field Index (DS) and Length-Field-Job Index (DSM). The specialist's evaluation allowed to establish the occupational cancer origin. The analysis of the indexes' accuracy showed good operating characteristics with higher specificity and equal sensitivity for DSM in comparison to DS. Therefore the short structured interview should be considered as a valuable tool to identify urothelial carcinomas worthy of further evaluation of occupational specialist. PMID- 18409704 TI - [Rubber industry and exposure to carcinogenic chemical agents: an Italian Multicentric Study in Lombardy]. AB - Evaluating the chemical risks at the workplaces, the potential carcinogenic risk is a primary interest. Particularly, in the rubber forming industry the main attention was directed to the hot processing as a possible source of exposure to chemical agents, several of which were classified as carcinogens by the International Agency for Research on Cancer (IARC) and/or the European Union (EU). A study on occupational exposure to chemical carcinogens in the rubber forming industry was planned during last two years, involving a representative sample of firms. In the first step the occupational exposure and the use of preventive measures were evaluated by an investigation at workplace supported with standardized questionnaires. The presence of above 100 chemical agents was registered; 15 agents were classified to carcinogenic or probably carcinogenic to human. The evaluation of exposure by air sampling and monitoring of most substances of interest (acrylonitrile, 1,3-butadiene, styrene, Polycyclic Aromatic Hydrocarbons), demonstrated a low level of occupational exposure, the common hygienic prevention measure being applied. PMID- 18409705 TI - [Plastic industry and exposure to carcinogenic chemical agents: an Italian Multicentric Study in Lombardy]. AB - The potential carcinogenic risk at the workplaces is a primary interest of occupational health, but some questions are also controversially discussed. Particularly, in the plastic forming industry a great attention was directed to the hot processing and their possible exposure to monomers, some of which were classified as carcinogen by the International Agency for Research on Cancer (IARC) and/or the European Union (EU). In Lombardy, a study on occupational exposure to chemical carcinogens in the plastic forming industry was planned during last years. The aim was to recognize and promote preventive technical and medical solutions, basing on efficacy. By an investigation at workplace supported with standardized questionnaires, the presence of chemical carcinogens was registered in 59% of a representative sample of firms; but an effective possibility of exposure was found only for 34% of cases. The evaluation of exposure to monomers by air monitoring (acrylonitrile, 1,3-butadiene, styrene, formaldehyde), involving a representative sample of factory with ABS and formaldehydic resins processing, showed low level exposure, because the common hygienic prevention measures were applied; some particular occupation shoved greater exposure to formaldehyde. PMID- 18409706 TI - [Prevention of road accidents in the road haulage field]. AB - Every year many traffic accidents with fatal outcomes occur in our Country. According to the recent indications of the European Agency for Safety and Health at Work, the Piedmont region has financed the plan: Prevention of road accidents in the road haulage field. OBJECTIVES: The aims of the plan are to stimulate transport companies to the target of road safety and to improve and enforce sanitary surveillance, in order to improve the safety on road haulage and to prevent traffic injuries. METHODS: the plan foresees, over a period of two years, a few encounters with all the interested parties (companies, police forces, labour unions etc). During those encounters we have to give a questionnaire for evaluating the companies' knowledge about the problem and we have to choose a common plan with the aim of improving road safety. RESULTS AND CONCLUSIONS: The Piedmont regional plan recalls the need to increase the attention to numerous and diversified hazards for safety on road haulage. It also imposes the choice of measures that include: risk assessment, health education, technical and environmental prevention, sanitary surveillance and clinical interventions (diagnosis and rehabilitation of occupational accidents). PMID- 18409707 TI - [The enforcement of the new Italian law on alcohol assumption in work activities with high accident risk in Lombardy]. AB - The 125/2001, the national law that forbids the alcohol consumption by workers with work tasks having high accident risks, has been activated a year ago. We carried out in Lombardy a research to evaluate the state of enforcement of the law in firms having different work activities. We found a complete application of the law only in very few firms. We think that the Occupational Health doctors should give more attention to the problem of alcohol consumption at the work places, giving the firms their specific scientific and medical support, using the peculiar knowledge of our discipline. PMID- 18409708 TI - [Commuting accidents: the influence of excessive daytime sleepiness. A review of an Italian Police officers population]. AB - Commuting accidents (CA) play an important role in many systems of workers' compensation insurance and with good reason, as they generally bring about more serious consequences in terms of permanent disablement and death than ordinary occupational accidents; this usually leads to high social costs. Nevertheless, research investigations aimed at studying the possible causes underlying the phenomenon are not available in medical literature. Objective of the present study is to evaluate whether excessive daytime sleepiness (EDS) might influence the occurrence of CA. 463 CA occurred to 411 police officers in northern Italy during the period 1999 - 2002 were collected; 51.9% of the subjects were working on shifts, 48.1% were non-shift workers. The study was carried out by submitting a self-administered questionnaire to gather information on age and physical characteristics, working conditions, sleep-related problems and accidents occurrence; EDS was measured by the Epworth Sleepiness Scale (ESS). A large number of workers (36%) complained of EDS; a strict significant relationship between shift - work condition and the presence of EDS was found, thus suggesting that CA are significantly influenced by EDS. The shift work schedule adopted by Italian Police might be accountable for the disruption of the balance between circadian and homeostatic factors. PMID- 18409709 TI - [Analysis of work accidents during the years 1999-2006 in a hospital company in Lombardia]. AB - This study describe accidents occurred in the period between 1999 and 2006 in the Hospital of Cremona, in which about 2400 subjects operate. The analysis of Accident Register showed a reduction of about 30% of the total number of accidents during the examined period and a non homogeneous distribution of the various types of accidents. The most frequent accidents were prick (25.8%), trauma (22.9%) and "in itinere" accidents (7.8%). One type of accident has been little considered up to now: the aggressions. Professional nurses were the most frequently involved and the most affected units were those that belong to the Internal Medicine Department. "In itinere" accidents had the longest average prognosis (11.6 days). The repetition of accidents occurred to the same operator hasn't been analysed before now: a professional nurse had nine accidents (of various type) in the seven years considered. Probably the reduction of accident must be attributed to the effectiveness of the prevention activities undertaken during the reviewed period. Biological accidents, for which it was possible to implement prevention programs, have been markedly reduced; it was not the same for "In Itinere" accidents, that depend significantly on external factors that are not easily dismissed. PMID- 18409710 TI - [Accidents in health care workers: the experience from the Careggi Teaching Hospital]. AB - In the last eleven years in the Careggi Hospital 4941 occupational accident has been registered, and 2951 workers were involved (28.9% of the total of the workers present in the period). The highest frequency of accident has been registered within the cooking staff, about one half of whom had an accident, and 7.2% more than 0.6 accident for person-year; the more serious consequences were registered for traffic accident, more than half happened coming to the workplace before the beginning of the work. Only 154 workers registered more than 0.6 accidents for person-year. PMID- 18409711 TI - [Work related mesothelioma: analysis of cases discovered at the Section for Occupational Medicine and Toxicology of Siena University during the years 2000 2007]. AB - This study focuses on the spread of mesothelioma in Siena. The population consisted of 30 patients. The diagnosis was made through histopathological and immunoistochemical or cytological and immunoistochemical analysis. The association between malignant masothelioma and exposure to asbestos was deduced by the occupational history. The mesothelioma was noted both in traditional industries and other jobs such as the chain of manifacture, plumbers, electricians, carpenters, installers of asbestos insulation and construction workers. Thus it is possible to find other malignant and nonmalignant asbestos related diseases more frequently than mesothelioma. There is an evident risk in rebuilding, so the development of new cases due to these exposures is expected. PMID- 18409712 TI - [Malignant mesothelioma (MM) in women: findings of the Mesothelioma Register of the Friuli Venezia Giulia Region]. AB - During the period 2000-2003, the Mesothelioma Register of the Friuli Venezia Giulia identified 248 cases of MM, 44 of which (18%) were female. In 36 cases the diagnosis was "certain" and in 8 "probable" or "possible". Mean age at diagnosis was 72.8 years (SD = 12.7), and the site of the disease was the pleura in 93% of cases. Information about previous exposure to asbestos was collected in accordance with the guidelines of the National Mesothelioma Register Occupational exposure to asbestos was documented in only 8 cases and family exposure in 6 others. In the remaining cases the source of exposure was "unknown" because of insufficient data, or there were no data at all. The study highlights the role played by extra-occupational exposure to asbestos among women and the need for careful investigation into previous asbestos exposure in all females with MM. In order to improve our knowledge of the part played by factors other than occupational exposure to asbestos in triggering the disease, it is crucial to reduce he number of cases with no information or "unknown" exposure to this dangerous substance. PMID- 18409713 TI - [Biomarkers for prevention and early diagnosis of malignant pleural mesothelioma]. AB - Improved detection methods for diagnosis of asymptomatic malignant pleural mesothelioma (MPM) are essential for an early and reliable detection and treatment of this disease. Thus, focus has been on finding tumour markers in the blood. 94 asbestos-exposed subjects, 22 patients with MM, and 54 healthy subjects were recruited for evaluation of the significance of 8-hydroxy-2'-deoxy-guanosine (80HdG) in white blood cells and plasma concentrations of soluble mesothelin related peptides (SMRPs), angiogenic factors (PDGFbeta, HGF, bFGF, VEGFbeta), and matrix proteases (MMP2, MMP9, TIMP1, TIMP2) for potential early detection of MM. The area under ROC curves (AUC) indicates that 80HdG levels can discriminate asbestos-exposed subjects from controls but not from MPM patients. Significant AUC values were found for SMRP discriminating asbestos-exposed subjects from MPM patients but not from controls. VEGFbeta can significantly differentiate asbestos exposed subjects from control and cancer groups. No diagnostic value was observed for MMP2, MMP9, TIMP1, TIMP2. The sensitivity and specificity results of markers were calculated at defined cut-offs. The combination of 80HdG, VEGFbeta and SMRPs best distinguished the individual groups, suggesting a potential indicator of early and advanced MPM cancers. The combination of blood biomarkers and radiographic findings could be used to stratify the risk of mesothelioma in asbestos-exposed populations. PMID- 18409714 TI - [Asbestos-related diseases in relatives of asbestos exposed workers]. AB - Standard asbestos diagnostic protocol was applied to eleven relatives of asbestos exposure ex-workers of a cement factory in Bari. Nine wives and seventeen sons were involved as volunteers in this evaluation. In this group two pleura malignant mesotheliomas (not dose-dependent) two asbestosis and fifteen pleura plaques (dose-dependent) were detected. This situation shows high level of asbestos contamination at home. For all the test patients the contamination most probably occurred because workers carried asbestos substances home from work on their clothes. PMID- 18409715 TI - [Evaluation of a series of serum mesothelin in patients with pleural malignant mesothelioma]. AB - Pleural Malignant Mesothelioma (MM) is a highly aggressive neoplasm with a poor survival rate, hard diagnosis and treatment. The incidence of MM in Western Europe countries is expected to increase drammatically in the next 10-15 years. In spite of this drammatic scenario, at this time the only instruments for screening and early diagnosis are based on radiological tests with evident ethical and economical problems. For this reason, some authors are evaluating biological indicators with the significance of screening and early diagnosis markers. One of the most promising marker is serum mesothelin (SMRP). SMRP levels appeares to be significantly related to MM and its clinical (diagnostic/prognostic) usefulnes has been suggested. The purpose of this research is to show SMRP trend in relation both to the course of the disease and the response to therapies in some Epithelioid MM patients. The analysis of SMRP levels in these patients suggests that it may be a useful marker for monitoring the response to treatment. In fact, it was observed that SMRP increases in patients who did not respond to therapy, it tends to remain stable when therapies results into a clinical stabilization, while it decreases after surgical procedure and in case of clinical improvement. PMID- 18409716 TI - [Serum mesothelin dosages in follow-up of previously exposed workers]. AB - High dosages of Serum Mesothelin have been demonstrated to be significantly associated to Pleural Malignant Mesothelioma. We recently demonstrated that Serum Mesothelin may be clinically helpful both for diagnostic and prognostic purposes, with the best cut-off corresponding to 1 nM. We also discovered that high levels of Serum Mesothelin are significantly associated to Lung Cancer. The usefulness of this marker in secondary prevention has been suggested, though never demonstrated. We therefore started a long-term prospective cohort study including previously asbestos-exposed workers. These subjects periodically underwent both radiological tests and serum mesothelin dosages. As a mid-term goal of this longitudinal study we decided to check the variability of mesothelin dosages, comparing baseline and follow-up values, as well as the possible correlation with age, duration of exposure, smoking, any abnormality of respiratory functional tests (RFT) and/or radiological tests. At baseline, Mesothelin mean value was 0.66 +/- 0.4 (range 0.08-2.2 nM). Both age (p = 0.04) and abnormal thoracic TC (p = 0.04) were significantly correlated with increased serum mesothelin levels and increasing age. No association was found between baseline mesothelin levels and duration of asbestos exposure (p = 0.5), smoking habits (p = 0.2), abnormal RFT, DLCO (carbon monoxide diffusing capacity) or thoracic X-ray. No significant variation was observed between mesothelin values at baseline and at follow-up (p = 0.2). PMID- 18409717 TI - [Medical surveillance of previously asbestos-exposed workers: report of a case of lung cancer with high level of serum mesothelin]. AB - Recently, the number of previously asbestos-exposed workers performing, at our department, medical exams aimed at an early diagnosis of asbestos-related tumors, has been progressively increasing. The diagnostical protocol we propose to these subjects include both radiological exams and some serum markers such as mesothelin and osteopontin. In this case-report we illustrate the history of a worker who, after having diagnosed a pulmonary asbestosis, developed a Lung Cancer. The significance of this case is based on the importance of the high mesothelin dosage which prompted further radiological exams resulting into the final diagnosis. In spite of the early diagnosis and treatment the patient finally died. Nevertheless, serum markers like mesothelin and osteopontin (especially the first) may result very helpful in monitoring and screening the population of workers previously exposed to asbestos. PMID- 18409718 TI - [Mortality from pleural and peritoneal cancer in a cohort of asbestos workers, many years after start of the exposure: possible role of fibers clearance]. AB - The multistage theory of carcinogenesis assumes rates of mesothelioma increasing monotonically as a function of time since first exposure (TSFE) to asbestos. However, some authors have suggested that the increase in mesothelioma rate with TSFE might be attenuated by clearance of asbestos from the lungs. We estimated mortality time trends from pleural and peritoneal cancer in a cohort of 3443 asbestos-cement workers. The role of asbestos clearance was explored using the traditional mesothelioma multistage model, generalized to include a term representing elimination over time. We observed 139 deaths from pleural and 56 from peritoneal cancer during the period 1950-2003. The rate of pleural cancer increased during the first 40 years of TSFE and reached a plateau thereafter. In contrast, the rate of peritoneal cancer increased monotonically with TSFE. The model allowing for asbestos elimination fitted the data better than the traditional model for pleural (p = 0.02) but not for peritoneal cancer (p = 0.22). The risk for pleural cancer, rather than showing an indefinite increase, might reach a plateau when a sufficiently long time has elapsed since exposure. The different trends for pleural and peritoneal cancer might be related to clearance of the asbestos from the workers' lungs. PMID- 18409719 TI - [Exposure to asbestos dust in agricultural environment. Some significant exposure forms documented by the Hospital Unit of Occupational Medicine of Hospital Institute of Cremona]. AB - Although it is difficult to document the exposition to asbestos in the agricultural workers, mesotelioma has however been noticed in the rural environment that may seem to have low risk because of its nature. This work describes the systematic diffusion of asbestos dust in the rural environment near Cremona due to the activity of "non-agricultural" workers who have fallen ill with mesothelioma. The patient described their work in details to the occupational medical doctor underlining the circumstances and the ways asbestos dust has been diffused in the rural environment. These ways were carefully analysed by the occupational medical doctor. They explain the onset of the mesothelioma in patients and they prove a systematic and long contamination of the involved rural areas. The described diffusion of dust has probably provoked a noteworthy exposure to the agricultural workers. At the moment there aren't full blown asbestos-linked pathology but these workers are potentially at risk of getting them. So it has an importance both on individual and global level for the rural area near Cremona. PMID- 18409720 TI - [Fast stress evaluation in nurses]. AB - The aim of the study was to assess subjective stress to 92 nurses by administering the Rapid Stress Assessment (RSA) Scale, a self-rated tool. The self-administered test, easily understandable, which is quickly drawn up and which allows a quick scooring, appears to be suitable to select subjects deserving a thorough specialistic investigation. The test itself becomes therefore a useful professional means for the occupational health physician, who has to evaluate the workers' psychic and physical attitude. PMID- 18409721 TI - [The evaluation of work related stress in occupational health: diagnostic experience and therapeutic prospects]. AB - The clinical cases of psychosocial disturbances that the Occupational Doctors find in their daily activity frequently are characterized by difficult differential diagnosis. These cases frequently must be faced with limitations of the work ability not easy to define and with unclear prognosis too. A useful help in the evaluation of these cases can be offered by the Clinical Units for the evaluation of work related stress, for the specialized joined support of Psychologist and of Occupational Doctors these Units can assure. We report the protocols of diagnosis of our clinical unit and the main characteristics of the patients more recently evaluated. PMID- 18409722 TI - [Analysis of a case series of workers with mobbing syndrome]. AB - Mobbing represents nowadays a major challenge for Occupational Medicine. We examined, during the last seven years, 253 patients who asked medical assistance for psychopathological problems by them ascribed to mobbing in the working environment. All patients underwent occupational health visit, psychological counselling (including personality tests administration), and psychiatric evaluation. A clinical picture probably due to mobbing was diagnosed in 37 workers: 2 cases of Post-Traumatic Stress Disorder (PTSD), 33 of Adjustment Disorder (AD), and 2 of anxiety disorder. Regarding mobbing typology, we found 19 cases of vertical mobbing (by an employer/manager to employees), 14 cases of strategic mobbing, 3 cases of horizontal mobbing (among colleagues), and one non intentional mobbing. In conclusion, a pure mobbing syndrome was diagnosed in a lower proportion than that reported by other investigators. The described interdisciplinary approach appears useful for the diagnostic assessment of suspect mobbing cases, that in turn is crucial for prognosis and treatment, as well as in relation to medico-legal issues and work-related compensation claims. PMID- 18409723 TI - [Study of a population exposed to occupational stress: correlation among psychometrics tests and biochemical-immunological parameters]. AB - A longitudinal study was carried out to evaluate the effect of psycho-physical and occupational stress on some biochemical and immunological parameters. The study was aimed to the identification of new and reliable method for the identification of subjects at high risk of occupational stress. 101 nurses which were working at several departments were enrolled. A blood sample was collected from all subjects after have filled the questionnaires at the time T0 and at the followed time points of 4 months (T1), 8 months (T2) and 12 months (T3). The self reported questionnaires were: Rating Scale for Rapid Stress Assessment (VRS), General Health Questionnaire to 12 items (GHQ-12) Multidimensional Scale of Perceived Social Support (MSPSS) and a questionnaire on the occupational satisfaction (SOD). Haemachrome glycaemia, homocysteine, cortisol, lymphocyte numbers and their subtypes (CD4, CD8, CD19, NK CD56, NK CD57), NK activity and inflammatory cytokines were evaluated. A high reliability has been found between the psychometric tests. Correlations between biochemical and immunological variables were performed by Pearson coefficients and multiple regression analysis. Subjects with elevated value of stress evaluated as VRS and GHQ-12 score showed an altered immune response. A reduction of NK CD57 and IL-6 content better characterize the occupational satisfaction. PMID- 18409724 TI - [Sex specific differences in physiologic response to stress evaluated by means of salivary cortisol]. AB - The psychophysiological mechanism behind the development of stress-related diseases includes a long-term both increase and decrease in circulating cortisol levels, leading to an allostatic disregulation of the hypothalamic-pituitary adrenal (HPA) axis. This research explores the relationship between perceived stress (assessed by means of the Job Strain Model) and neuroendocrine response quantified by means of repeated measures of salivary cortisol in 46 call-centre operators. Job strain influenced the total amount of cortisol response to waking, but not the cortisol excretion in the remainder of the day. The cortisol response to waking showed gender-specific differences, women excreting greater cortisol than men [AUC(t): coeff (IC 95%) = 16.2 (5.3-27.1); AUC(i): coeff (IC 95%) = 8.3 (2.4-14.2); MnInc: coeff (IC 95%) = 5.2 (1.6-8.9)]. In long run the gender specific differences of the dis-regulation of the hypothalamic -pituitary-adrenal (HPA) axis can be related to differences on prevalence of autoimmune diseases. PMID- 18409725 TI - [Risk assessment method for psychosocial factors at workplace: a North-West Tuscany area project]. AB - The rising awareness of psychosocial risks at workplace means that it is ever more important to prepare methods to assess psychosocial factors in occupational environment. This project of north west tuscany area has the aim to realize an instrument for a gradual risk assessment for this kind of factors without the support of specialists. A decisional flow chart helps to approach the risk assessment step by step on the basis of company features, management and organization problems and company symptoms of stress. The final assessment combines the evaluation of perceived risks with job analysis realizing a matrix containing 5 risk levels, which suggests the priority of preventive measures. The intermediate level (level three) represents the activation level for the medical surveillance. This experience means a proposal for a quantitative assessment of psychosocial risks at workplace. PMID- 18409726 TI - [A method for the measurement of workplace violence in health care workers]. AB - Aiming to register workplace violence in health care workers, we adopted two evaluation methods: direct and indirect. The direct method is represented by the Violent Incident Form (VIF), a 16 items checklist, which resumes the key aspects of violence events, identifying spatial and temporal circumstances, aggressor, type of violence and consequences. Indirect method is constituted by the observation of injury reports, through three sources: the Prevention and Protection Service (SPP), the Public Relation Office and the Police Position of the First Aid Unit. Although the instruments adopted, thus not comparable each other, pointed out a rather high number of violence events suffered by health care workers (direct method: 34 reports in the period 2002-2006; indirect method: 25 subjects, 8.5% of total); in most cases, the events were perpetrated by a patient, and directed against female of nurses. The most reported types of violence were: kicking, beating, hitting (indirect method) and verbal assault/threats (direct method). The study pointed out that workplace violence is an important risk factor, and therefore it is necessary to consider it for preventive intervention. PMID- 18409727 TI - [Cardiovascular and hormonal strain in waste collection workers employed in Amsa SpA, municipality of Milan, in permanent day and night work]. AB - Aim of the study is to explore in shiftworkers: a) blood pressure and hormonal variations; b) dyslipidemia and blood glucose levels; c) insulin resistence syndrome. We have assessed 48 male workers employes in Amsa SpA, a large municipal enterprise in charge of street cleaning and domestic waste collection, in permanent day and night work as hand sweepers, motor sweepers and delivery tricar drivers. 24 of those workers (daily and nightly) were normotensive and 24 were hypertensive. Our medical checks were: physical examination: BMI; laboratory findings (blood): glucose, total cholesterol, triglycerides, endothelin, insulin, FFA, HOMA S, HOMA B, HOMA R; assay of salivary cortisol and urinary cortisol in 24 h; 24 h pressure monitoring. RESULTS: Nightly hypertensive: increased consumption in wine and coffee, weight and BMI, total cholesterol and FFA and endothelin. Nightly normotensive: increased consumption in cigarettes and salivary cortisol. Daily hypertensive: increased total cholesterol; 24 h pressure monitoring showed more pronounced variations of pressure in night workers both normotensive and hypertensive in working time. CONCLUSIONS: Night shiftwork looks like more stressfull than day shiftwork. Insuline resistance isn't noticed in all four groups. PMID- 18409728 TI - [Algorithm validation for pesticide exposure risk evaluation by environmental monitoring data]. AB - The risk evaluation related to the occupational exposure to hazardous substances can be performed either by analytical data coming from environmental and/or biological monitoring or by using models and mathematical algorithms. Models are often considered economically more advantageous, nevertheless, in order to assure their reliability, they need to be validated by objective monitoring data. A predictive model for the evaluation of risks due to the exposure to pesticides in agriculture was elaborated, and, for model validations, 20 tomato farms were examined. For each farm, information necessary to the risk evaluation by using the elaborated algorithm were collected and, simultaneously, an environmental monitoring for the evaluation of inhaling and cutaneous exposure was carried out. The potential risk was estimated by comparing analytical data from environmental monitoring with respect to ADI (Admissible Daily Intake) values, for each investigated pesticide. Then, the accordance between the potential risk and the risk given by the elaborated model was evaluated and an agreement of 85.6% was found, showing a substantial reliability of the proposed model. PMID- 18409729 TI - Risk characterization and proposal of exposure thresholds to indoor chemical agents: the Index European project. AB - The European INDEX project was finalised to identify priorities and to assess the needs for an European Community strategy and action plan in the area of indoor air pollution. A list of 5 chemicals, with potential of high indoor concentrations, uncontested health impacts, and effective risk management, were selected to be regulated with priority: Formaldehyde, Carbon Monoxide, Nitrogen Dioxide, Benzene and Naphtalene. For each compound, guideline values and management options were suggested. PMID- 18409730 TI - [Model of chemical risk assessment (D.L 25/2002), applied in 19 chemical industries of the Province of Bergamo]. AB - The aim of this work is to describe a model for the evaluation of chemical risk, according to the legislative decree 25/2002, planned by U.O.O.M.L. "Ospedali Riuniti di Bergamo", in collaboration with "Gruppo Industriali Chimici dell'Unione Industriali-Provincia di Bergamo" and applied in 19 Chemical Industries of the Province of Bergamo. The objective of the model is the rationalization of the procedures to ponder the levels of exposure and to value different typologies of risk using and manipulating chemical substances with the purpose to optimize the relationship among the attention for safety, workers health and the employment of resources. PMID- 18409731 TI - [Risk perception and representation in construction sites: a preliminary study]. AB - The number of building injuries and professional health disease make evidence of safety problems in this field. The study of human factors may help us in observing this problem in a better way (more global). The aims of this study is to investigate construction workers risk perception and representation. Which are the risk? Which risk is perceived as more dangerous? What is Risk?. These are some question this study try to give an answer, asking about worker perception and representation of risk. Qualitative analysis shows which are the meanings workers use when they talk about risk, underlining the influence of personal, group, organizational and cultural variables. PMID- 18409732 TI - [Evaluation and management of pesticide risk in agriculture: the experience of the Region of Lombardy (Italy)]. AB - The evaluation of chemical risk in agriculture is complicated because of difficulties in obtaining measures representative of working conditions. This is the reason why experiences finalized at producing risk estimates are running. In this frame, a Regional working group has developed the project "Pesticide exposure and risk profiles in agriculture". Priority scenarios have been selected and the main variables correlated with pesticide exposure have been pointed out. A value for each variable has been defined. The sum of these values allows the definition of "Exposure Indices" (EI), which can be reduced by multiplication for a coefficient calculated based on use of personal protective devices, training and education and equipment conditions. A Risk Index is calculated as the product of EI per a toxicity index, calculated based on the risk phrases of the substances used ("Risk Profile"). Risk Profiles allow the production of risk estimates and the definition of the appropriate preventive interventions. Next phase will be addressed at the validation of the model, to be carried out through the determination of the levels of concordance between the risk class allocation obtained from the model and the one obtained from environmental and biological measures, in the same groups of workers. PMID- 18409733 TI - [The impact of personal protective equipment in reducing risk for operators exposed to pesticides: from theory to practice]. AB - In Europe, the use of plant protection products is authorized provided the operator risk assessment carried out does not show exposure levels above the health-based triggers. Two models are currently available (German and UK models) to estimate exposure levels of agricultural operators. They consider, among different variables, the use of Personal Protective Equipment (PPE), based on their capacity to reduce exposure through the application of specific "reduction factors", derived from laboratory or field studies. Such factors, which date back to '90s, could be obsolete, as indicated by recent literature, posing problems on the reliability of current estimates. Therefore, the exposure estimates produced in Europe from 2005 to 2007, for 52 active ingredients in 395 scenarios, have been collected and analysed to check whether the application of recent exposure reduction factors might have an impact on the final outcomes. In some case the exposure levels resulted underestimated, highlighting the problem of operator safety with regard to the use of specific PPE and to an adequate training programme. PMID- 18409734 TI - [Chemical pollutants "exposure assessment" experiences: mathematical modelling vs environmental monitoring]. AB - This contribution presents the employment of two risk estimation mathematical modelling--InfoRisCh and MoVaRisCH--in the context of an exposure assessment updating for different dangerous products in a petrolchemical facility, and therefore the comparison with occupational safety team and environmental monitoring results. PMID- 18409735 TI - [Ten years of protection of the maternity leave of women working in Pistoia]. AB - We have analyzed 1175 interventions about pregnant women who came, from 1997 to 2006, to our public occupational health service, asked for the advance the abstention of work during pregnancy. Requests for advance the abstention of work during pregnancy came from service sector (welfare, school and cleaning--29%), shoe industry (14.4%), textile and clothing industry (11.4%), commerce (characterized by prolonged standing straight 13%), health service (8.7%), tourism (8%). The first seven years (1997-2003) the prevailing risks have been the muscular effort and the postures, while the last three years (2004-2006) have increased biological and chemical risks. The most important principles of labour protection for pregnant women is the agreement with every subject, and the information of the social parties, but above all the direct information of the women. PMID- 18409736 TI - [Sick leave risk factors in a sample of women employed in the public sector. The SEMM study]. AB - OMS reports describe an increase of levels of sickness leaves (SL) at work in all industrialized countries in last decades. Aims of this study are to assess incidence of SL from work in a sample of women public employees and to identify significant factors, among individual, behavioural, familiar, occupational and work strain related. The survey took place between April 1992 and December 1998. After the baseline examination, a two years follow-up was conducted, to detect any SL longer than one day. N. 8123 female employees of the Municipality of Milan were enrolled, undergoing a baseline health examination. They belong to the following municipality sectors (SM): kindergarten, register office, clerks and police. The overall response rate was 76% (3698 females and 2448 males). The present analysis was restricted to female only, who showed almost double incidence of SL than men. Interpolation of logistic models allowed to identify the following predictive independent factors: physical activity in leisure time, work length and social support at work, for incidence of short SL; physical activity at work, hypertension and smoking habit for intermediate SL; and perceived work strain and number of familiar contacts for long SL. PMID- 18409737 TI - [Emerging problems in enforcement of safe maternity and feeding protection at work: a public prevention service experience]. AB - We have examined the claims for advance maternity leave or prolonged benefits for breastfeeding addressed to Occupational Health Unity of Local Health Service 11 by women at harmful works in the period 2002-2005. The most frequent occupations were: shoemaker (29%), service company's employee (7%), tanners (7%), leather industry's employee (6%) and food industry's employee (6%). The most important risk factors were: bound postures, manual load handling, chemical hazards and biological agents. The numbers of claims increased during the period of interest. In the work place often risks for pregnancy and breastfeeding are not correctly assessed and women workers are not informed on their rights. The Occupational Health Unity of Local Health Service 11 tried to correct the lack of information for workers, employers, workers' representatives in health and safety and enterprise's occupational health physician. PMID- 18409738 TI - [Injuries from violence in the hospital: assaults in the psychiatric unit]. PMID- 18409739 TI - [Health status of women at work: work risks and living conditions]. AB - The increase of women at work calls for a new attention to a full health protection, besides the fertility and reproduction. Health Surveillance in workplaces can give much information about health state of women and men, when the evaluation takes into account physiological and social differences between the sexes. The study reports the health data from a working population, 675 women and 7991 men, employed in different work activities. The results showed no significant difference of health state between women and men, except a greater prevalence of the respiratory pathologies in men and psychosomatic disorders in women. Prevalence of muscle-skeletal diseases, psychosomatic disorders and recurring headache have been higher in married than in unmarried women. among married women, prevalence of pathologies have been related to number of children. No difference have been found between unmarried and married men, except a greater prevalence of psychic disorders in youngest. Results confirm the interaction between domestic and working load on health state of women. Under the same work conditions, women are subjected to a higher physical and mental load that reduces the endurance of strain and stress and increases the prevalence of some pathologies, as musculoskeletal chronic degenerative diseases and psychological disorders. PMID- 18409740 TI - [Work and depression in women after the birth of a child]. AB - OBJECTIVE: To analyze the main factors associated to depression at 8 months postpartum, looking more particularly at women's employment. METHOD: 352 women responded to a face-to-face questionnaire few days after the birth, at the Maternity Hospital "Burlo Garofolo" in Trieste, and 292 of them responded to a telephone interview 8 months later. Psychological distress 8 months after delivery was evaluated with the General Health Questionnaire (GHQ). RESULTS: 8 months postpartum, 5% of women were depressed. 32% of women were non-satisfied with their current working status, without differences among those working and those staying at home. 13% of sample reported problems related to work (such as being fired or no flexible working hours). The women's actual working status was not associated with depression, while the congruence between what the woman was doing (working/at home) and her wishes was. The relationship was still significant after adjusting for other factors associated with depression. CONCLUSIONS: Employment dissatisfaction is negatively associated with women's health after childbirth. To promote mothers' wellbeing, the question of women's employment after birth should be addressed. PMID- 18409741 TI - [Pregnancy and work in the area Livorno]. AB - This document describes the observatory on working women in Livorno area. The sources of information are 1) individual information sheets collected from midwives with a specially designed questionnaire for all pregnant women from 1/4/2003 to 30/12/2006 2) data from the procedures of interdiction for risk work. The analysis of data sheets shows the following information: 9% of pregnant women are foreign and 70% are workers. The sectors where women are mainly employed are the following: Administrative (34%), Commerce (17%), Health (10%), Cleaning (6%), School (4%), Metal Industry (3%), Hair stylists (2%), Food (2%). The analysis of data by procedure of interdiction for risk work shows that the most represented sectors are: Commerce (20%), Health (14%), Cleaning (22%), Metal Industry (21%), School (12%, Hair stylists (8%) and Food (3%). There is an evident increase in the number of women who come to our occupational health service and in the number of interdiction of risk work. However there are still problems in some critical areas with predominantly female workforce (cleaning, commerce) and in case of foreign workers and women with insecure contracts. PMID- 18409742 TI - [The role of occupational physician in the application of the 2006 "agreement on workers' health protection through the good handling and use of crystalline silica and products containing it": the experience in mining sector]. AB - The 2006 "Agreement on Workers' Health Protection Through the Good Handling and Use of Crystalline Silica and Products Containing it" between social parts defines a standardization of exposition control methods and medical surveillance. The Occupational Physician is integral part in exposition evaluation process and risk stratification in which derives the medical surveillance program. This study presents a first application of the European agreement in mining sector and the role of Occupational Physician in the evaluation of the risk to define methods of prevention. In particular it will be precised the choice of homogenous groups, the classification of exposed workers from results of workplace monitoring, the choice of technical prevention and individual protection equipments, and then the strategy of medical surveillance. PMID- 18409743 TI - [Crystalline silica can induce oxidative stress by inhibiting glyoxalase system in bronchial epithelial cells]. AB - Chronic inflammation and reactive oxygen species (ROS) production induced by crystalline silica are involved in the development of silicosis and lung cancer pathogenesis. ROS can generate lipid peroxydation of cell membranes that can produce methylglyoxal (MG), a strong cell proliferation inhibitor and apoptosis inducer. MG is naturally removed by glyoxalase I (GI) and glyoxalase II (GII) through a glutathione (GSH) dependent mechanism. Therefore mRNA expression of glyoxalases is correlated to MG concentration and oxidative stress. OBJECTIVES: evaluate oxidative stress induced by crystalline silica by glyoxalases mRNA expression and methylglyoxal concentration MATERIAL AND METHODS: In bronchial epithelial cell culture (BEAS-2B), exposed to 50 microg/cm2 crystalline silica (Min-U-Sil 5), for 2, 6, 12, and 24 hours, GI and GII mRNA levels and MG intracellular concentration were measured respectively by Real-Time PCR and HPLC. RESULTS: Crystalline silica exposure induced a significant reduction in mRNA expression of glyoxalases and an increase of MG intracellular concentration. CONCLUSIONS: The results suggest a possible use of MG and mRNA expression of GI and GII as crystalline silica induced oxidative stress indicators. PMID- 18409744 TI - [Role of screening tests for indirect diagnosis of tuberculosis in health care workers: Mantoux and the new tests on blood ELISA]. AB - The Tuberculosis infection in recent years has become always more a threat. The failure in the attempt to stop it (O.M.S. Millennium Global Plan) brought to the revision of the world control strategy to at least contain this disease (The Global Plan to Stop TB 2006-2015). Due to these severe facts it is even more important now to elaborate more sensitive and specific methods to find out, as fast as possible, the infected cases. As of today, the main TB infection screening test is the Skin PPD test (Mantoux). Recently new tests for the population screening are in use; these tests are based on the evaluation of immunity cell-mediated. They (QFT-G) do not have the typical limits of the Skin Test and they are more suitable as serial tests and therefore more useful, according to us, in the screening programs of the TB infection in low prevalence countries, like Italy. PMID- 18409745 TI - [Emerging infectious diseases among swine workers]. AB - In the last years emerging infections represent an important problem of public health and occupational medicine. Biological agents and their hosts exist in a precariously balanced and continuously evolving relationship, influenced by their environment. RNA viruses are responsible for most of the emerging diseases. Epidemics that recently affected the world of work are zoonoses, such as cases of SARS in healthcare staff Dutch poultry workers infected with the avian virus A/H7N7 in 2003, the current threat of avian flu A/H5N1 to poultry workers. Workers at risk include those who are in contact with live or dead infected animals, with aerosols, dust or surfaces contaminated by animal secretions, persons engaged in animal breeding and trade, veterinaries, and others. Pigs are at risk of acquiring many viral and bacterial diseases and, consequently, could be able to transmit some of these infections to occupationally exposed subjects. The aim of our study is to set out some emerging zoonosis that could affect swine workers, an occupational sector where a proper assessment of biological risks is difficult to perform. PMID- 18409746 TI - [The risk management in health personnel tuberculosis]. AB - Within work environment and even more among health service areas, to deepen the monitoring efforts and awareness of the workers in order to accomplish a more effective and fast response against morbidity indexes and prevalence of tbc, constitute the necessary prerequisite for any subsequent action of prevention and control. On the other hand, one of the employer's obligations toward a health facility is to identify professionally exposed subjects (Legislative Decree 626/94 art.78), and perform risk assessment. Continuous report from various hospitals concerning suspicious contacts between employees and patients affected by tbc further increase the threat. At AOUP in Palermo from 2003 to May 2007, 39 cases of suspected contact between employees and patients affected by tbc were reported to the 'Staff Union for Health Related Issues' by various hospital departments and the 'Office of the Medical Competent', 34 of these reported cases have pulmonary involvement. The above mentioned situation made necessary to launch an effective program capable of addressing the risks of tbc in both circumstances of suspected or certain contact and during normal work activities. PMID- 18409747 TI - [Screening of latent tuberculosis infection in health care workers by QuantiFERON TB and tuberculin skin test]. AB - Recent guidelines (MMWR 2005) recommend the use of QuantiFERON-TB (QFT-TB) as an alternative to the tuberculin skin test (TST) for the screening of latent tuberculosis infection (LTBI) in health care workers (HCWs). MATERIALS AND METHODS: 590 HCWs were screened for LTBI by TST and QFT-TB. Results were compared with risk factors for LTBI, determined by questionnaires. RESULTS: Both tests were significantly associated with non-Italian nationality [TCT (OR = 9.17), QFT TB (OR = 3.65)], age > or = 45 years old [TCT (OR = 1.81), QFT-TB (OR = 2.36)], history of household contacts [TCT (OR = 2.65), QFT-TB (OR = 3.37], occupational exposure to tuberculosis (TB) patients [TCT (OR = 2.14), QFT-TB (OR = 1.93)]. 55 cases were discordant (28 QFT-TB-negatives/TCT-positives; 27 QFT-TB-positives/TCT negatives). Both tests were not associated with workplace risk factors or TB risk level assessed in different hospital units. CONCLUSIONS: In HCWs employed in a low TB incidence area both QFT-TB and TCT were more associated with non occupational risk factors (nationality, age, household contacts) than with main determinants of workplace risk for LTBI. PMID- 18409748 TI - [Susceptibility to varicella-zoster, measles, rosacea and mumps among health care workers in a Northern Italy hospital]. AB - BACKGROUND: Nosocomial transmission of varicella (V), rubella (R), mumps (Mu) and measles (Me) may be a significant cause of morbidity in health care workers (HCW). Susceptible HCW might be a relevant part of the workforce. METHODS: A seroprevalence study was performed in a public hospital. Antibodies (Ab) against V, R, Mu and Me were determined by ELISA. Sociodemographic, clinical, occupational data and sera were obtained during health surveillance. RESULTS: 2934 tests on 1106 HCW were performed. Seropositivity was 91% for V, 89% for R, 80% for Mu, 92% for Me. No significant differencies were found for the variables tested, except for age (< 36 years) and certain job tasks (e.g. residents). 22% of HCW tested were seronegative for at least 1 virus. More seronegatives were detected as compared with the majority of literature data. DISCUSSION: This study showed a relevant number of susceptible to V, R, Mu and Me. Seroprevalence data are useful for risk assessment, HCW health surveillance, to evaluate fitness for work and to promote vaccination programmes, according to scientific guidelines. OP should include serological screening for Me, Mu, V and R to protect HCW and third parties. PMID- 18409749 TI - [Tuberculin skin test (TST) survey among healthcare workers (HCWs) in hospital: a systematic review of the literature]. AB - OBJECTIVES: To summarize the evidence on the Tuberculin Skin Test (TST) survey among healthcare workers (HCWs) in hospital, in the last decade. METHODS: A systematic search was conducted in PubMed and in the Proceedings of the congresses of the Italian Society of Occupational Health and Industrial Hygiene (SIMLII); researches conducted in areas with medium-high incidence of Tuberculosis were excluded. FINDINGS: 25 publications were identified. The compliance of HCWs to TST surveys ranged from 80 to 100%, the incidence of TST conversion from 0.07 to 3.7% and the prevalence of positive TST from 5,7 to 57%. TST-conversion and positive TST were significantly associated to BCG vaccination and foreign-born. Most studies found an association between TST-conversion and occupational exposure in certain location (emergency facilities, internal medicine, infectious diseases, intensive care units, pathology, microbiology) and occupational categories (nurses, laboratory technicians). In some studies, period of hospital employment do not increased TST conversion rate. DISCUSSION: Positive TST and TST conversion rate seem to be associated with occupational exposure to Mycobacterium Tuberculosis in hospital, considering non-occupational factors. Evidence from literature seemed to support the use of TST in the tuberculosis screening programme of HCWs in hospital. PMID- 18409750 TI - [Anesthetic gases exposure: findings from a 13 year environmental and biological monitoring in a hospital company]. AB - The evolution of the modern anaesthesiologic technique aimed to detect more manageable and less toxic aesthetic gas, but adequately efficacy. This study considered the environmental and biological evaluation of the Careggi Hospital in Florence between 1991 and 2005. The pollution threshold of each aesthetic gas used (Nitrous oxide, Isoflurane and Sevoflurane) is reduced as time passed. The higher values about the pollution threshold have been found in the ophthalmology and in the otolaryngology operating room. We observed clinical and biological data of 74 health care workers (HCW). The anaesthetists and assistants were the more exposition. We didn't found any correlation between the exposure at gas and the clinical, and biological data. Some neurological and nonspecific symptoms (like head-ache and reduction of attention threshold) appear from the clinical informations; these symptoms suggest a possible effects on the HCW' health also for low pollution threshold. PMID- 18409751 TI - [Vaccination and immunization in healthcare workers, towards specific infections: measles, varicella, rubella and mumps. The experience in a hospital in Lombardy]. AB - Lombardy Region, with the Deliberation NoVIII (22-12-2005), about vaccinations in childrens and adults, suggest to offer to the healthcare workers (HCW) of 'Infectious diseases' and of 'Obstetrics and 'Pediatrics' Department, the vaccines for varicella and measles, mumps and rubella (MMR). We performed in 120 HCW of our hospital the dosage of antibodies versus these infectious diseases, in order to protect both workers and critical patients. The study results show that more than 80% of the HCW was immune to all the four infectious diseases. The percentage of immunisation to measles, varicella and rubella exceeded the 90%, while 87.5% of HCW was immune to mumps. We are going to offer the vaccine to the operators that are not immune, but we are also thinking about offer it to the HCW working with critical patient. PMID- 18409752 TI - [Latex risk prevention in San Paolo Hospital in Milan]. AB - Several epidemiologic studies have highlighted that latex sensitization prevalence rate has increased over twenty years both in the general and working population, mainly among health care workers. Such subjects can develop immediate local or systemic hypersensitivity reactions up to anaphylactic shock. First, at San Paolo Hospital in Milan, it has been determined latex sensitization and allergy prevalence rates in health care workers involved in health surveillance. Subsequently an interdisciplinary task group has been constituted in order to plan latex pathology preventive actions in health workers and to identify the preventive measures that must be applied in allergic patients. These facets are tightly one another linked. Since latex allergy primary prevention is the most effective, but difficult to put into effect. Operational protocols, by which recognize latex allergy risk factors and reduce exposure to this substance, have developed for both workers and users. Latex allergy and sensitization occurrence will not probably be erased by these procedures application, but they will be reduced within the limits as low as possible. PMID- 18409753 TI - [Prevalence of Staphylococcus aureus methicillin-resistant (MRSA) among health care workers]. AB - Methicillin-resistant Staphylococcus Aureus (MRSA) is a type of Staphylococcus that is resistant to certain antibiotics. These antibiotics include methicillin and other more common antibiotics such as oxacillin, penicillin and amoxicillin. Staphylococcus infections, including MRSA, occur most frequently among persons in hospitals and healthcare facilities. The present study was performed to investigate the in vitro activity of oxacillin and other antimicrobial agents against S. aureus strains obtained from nursing personnel. The study included 56 hospital personnel of Universitary Policlinic of Messina. S. aureus strain was isolated in 14 samples (25%); resistent patterns have been studied and results have demonstrated: none methicillin resistant, while 14% oxacillin and tetraciclin resistant. The incidence of methicillin sensitive was 100%, while 86% proved to be sensitive to oxacillin and tetraciclin. In conclusion, the usually hygienic methods (disposable gowns, hygienic hand disinfection after each patients contact, masks use when is a risk of aerosolization of MRSA) are indicate for significantly reducing of these strains. Continuing education programmes can help to increase awareness among hospital staff. PMID- 18409754 TI - [The environmental monitoring of the exposure to chemical contamination in operating rooms]. AB - The medical staff which works in an operating room is exposed to danger due to the chemical contamination found in the air. The results of this research depend to hormonal and haematochemical variations. The chemical contamination can be the cause of pathologies of the respiratory organs, the skin, the mucosa and the immune system. After a preventive evaluation of the production processes and the working procedures, some researchers have estimated the environmental risk caused by low concentrations of chemical products. In order to control the levels of the chemical agents, they have used an integrated system set up by a gaschromatography-mass spectrometer that has found some levels of chemical agents peculiar to a low pollution, characterized by the low concentration under the levels of the so-called ACGIH (2006) of hesane, toluene, ethylbenzene, o-xylene and naphthalene. According to the latest studies is very important to develop working methods and scientific knowledges direct to environmental, medical and toxicological problems. They are necessary to guarantee a greater protection of the human health and the safety at work. PMID- 18409755 TI - [Latex allergy in hospital workers--follow-up of symptomatic subjects]. AB - Latex allergy may be manifested in a variety of clinical disturbances such as contact urticaria, angioedema, rhinoconjunctivitis, asthma, and anaphylactic reactions. The aim of this follow-up study was to determine whether a change in glove use from powdered to powder-free latex gloves at previously surveyed hospital workers reduced the work-related symptoms. 53 hospital workers with work related symptoms where followed up 4,6 years after first investigation. On re examination, 54.7% use both powdered and powder-free gloves, 37.7% used only latex free gloves and 7.6% stopped the glove use. At follow-up, in 68% of subjects there was the remission or the improvement of work-related symptoms. The improvement of symptoms was greater in workers using powder-free gloves than in others (p < 0.005). In conclusion our study shows that preventive measures, such as the use a powder-free latex gloves, are sufficient to induce a reduction of work-related symptoms. PMID- 18409756 TI - [Evaluation of the genotoxicity of the extremely low frequency-magnetic fields (ELF-MF) in workers exposed for professional reasons]. AB - Whether or not ELF-MF has genotoxic potential is a controversial issue. In the present study, we investigated the genotoxic effect using cytogenetic assays (CA SCE - MN), in 109 workers exposed to ELF-MF. The mean value of occupational exposure in the whole group was 0.35 microT. According to the exposure level the workers were stratified in two different groups: low exposed (n. 39, TWA < or = 0.2 microT) and higher exposed (n. 70; TWA > 0.2 microT): the groups did not significantly differ for the examined variables. Due to these results we decided to re-evaluate the effect only considering the highest exposed workers: we selected 31 workers exposed to TWA levels exceeding 1 microT, and compared vs. the low exposed subjects: again, the difference in the groups examined were not significant. At multivariate regression analysis was also applied: no correlation was observed with cytogenetic biomarkers. The results of this study does not support the hypothesis of any direct genotoxic effect of ELF-MF, at least at the environmental levels currently found in an occupational settings. PMID- 18409757 TI - [Professional exposure to natural ultraviolet radiation: risk assessment and management and preventing strategies]. AB - The aim of our study was to verify the impact of work risk factor in causing cutaneous neoplasia on a group of patients and to assess the risk of exposure to UV on outdoor workers. The survey was divided in three phases: a) Questionnaire, b) Dermatological clinical examination. c) Exposure measurement by spectral radiance method (mW/cm2). 248 subjects were enrolled, 100 of which affected by basal cellular carcinoma (BCC) (68% M and 32% F: mean age 70 +/- 11.93 y) 130 by melanoma (42% M and 58% F; mean age 53 +/- 17.35 y) and 18 by squamous cellular carcinoma (33% F and 67% M: mean age 78 +/- 11.41 y). In addiction 22 healthy male subjects exposed to UV were examined (mean age 38.59 +/- 13.65 y; specific working age 11.88 +/- 10.92 y). The assessment of UV exposure was performed over a 24 days timeframe during summer 2007, all over the working day. Furthermore, maximum allowed exposure time (t(max)) without protection was calculated according to ACGIH guidelines in order to prevent cutaneous alterations. It turned out that the UV exposure, both in working and spare time, is the root cause for squamous cellular cancer developing. Furthermore our results proved that also BCC is related to the overall UV exposure. No clear link to the UV exposure was found for cutaneous melanoma. No precancerous lesions were observed on shortly exposed subjects. The measurements performed revealed, as expected, lower exposure during first and last hours of the day without any particular constrain on t(max), and maximum exposure in the timeframe between 10:00 AM and 4:00 PM. Therefore, it is necessary to pay higher attention to the UV exposure and related consequences on employees health in order to provide with the most suitable preventing measures such the usage of protections depending from the requested exposure time. PMID- 18409758 TI - [ELF-MF occupational exposure: repeated measures on workers]. AB - The aim of our study was to estimate occupational exposure to Extremely Low Frequency-Magnetic Fields (ELF-MF). Using personal dosimeters we evaluated occupational exposure in 543 workers employed in 9 occupational settings (about 150 jobs), representative of the main occupational activities in Emilia-Romagna region. In the whole sample, the median Time-Weighted Average (TWA) exposure resulted 0.14 microT (5 degrees - 95 degrees percentiles: 0.04 - 2.50 microT); Tile production presented the highest occupational setting TWA (0.46 +/- 1.83 microT), while the lower job TWA was Stock Clerk in Tile Production and Stock Clerk in Garment Production. After 6-9 months we repeated measurement in about 10% of the original sample (about 48 subjects) to verify the resulted obtained. At repeated measurement analysis with SPSS 12.0 no variation was found compared the previous monitoring. Individual occupational exposure to ELF-MF evaluated in a relatively large group of workers engaged in the main occupational activities proved low values compared to the proposed occupational limit. PMID- 18409759 TI - [Collaboration between occupational doctors in the public prevention services and qualified occupational doctors: description of an experience in the sandstone sector of Alto Mugello]. AB - BACKGROUND: The present course of work-related injuries imposes an effective collaboration among public service's doctors and qualified occupational doctors. We refer to a positive interaction in a prevention plan of hand-arm vibrations exposure during phases of dressing in the sandstone sector of Alto Mugello. METHODS: Both were acquired data on exposition (risk assessment, measures on tools, times and levels of exposition, etc.) and data on workers' health (an high number of workers showed suspect angioneurotic disorders). The symptomatic workers were subjected to a clinical-instrumental examination that highlighted a clear correlation between high hand-arm vibrations exposition and functional injury. RESULTS: Sharing informations is a precious tool both for Public Prevention Services, it enables in fact to realize an accurate representation of workers' health state, and for qualified occupational doctors to manage risk with employers and to carry out an effective health surveillance. PMID- 18409760 TI - [Regional planning of prevention measures in work environment: Veneto Region experience]. AB - In Veneto, from 1999, activities for health promotion in workplaces have been defined like a three - years planning, based on European, national and local policy, and on work accidents and injuries epidemiologist data. Activity plans have always been managed by using Plan - Do - Check - Act quality cycle, with in itinere and ex post evaluation steps to control system performances, and to define necessary activities for a never- ending improvement. PMID- 18409761 TI - [Implementation of a safety and health planning system in a teaching hospital]. AB - University Hospital "L. Sacco" had started in 2006 a two-year project in order to set up a "Health and Safety Management System (HSMS)" referring to the technical guideline OHSAS 18001:1999 and the UNI and INAIL "Guidelines for a health and safety management system at workplace". So far, the following operations had been implemented: Setting up of a specific Commission within the Risk Management Committee; Identification and appointment of Departmental Representatives of HSMS; Carrying out of a training course addressed to Workers Representatives for Safety and Departmental Representatives of HSMS; Development of an Integrated Informative System for Prevention and Safety; Auditors qualification; Inspection of the Occupational Health Unit and the Prevention and Safety Service: reporting of critical situations and monitoring solutions adopted. Short term objectives are: Self-evaluation through check-lists of each department; Sharing of the Improvement Plan among the departments of the hospital; Planning of Health and Safety training activities in the framework of the Hospital Training Plan; Safety audit. PMID- 18409762 TI - [Quality oriented education: the experience of the Prevention Department of the Empoli Local Health Company n 11]. AB - Quality management (QM) is a key issue for the Italian National Health System (NHS). Regione Toscana is currently undertaking a pilot programme to promote QM in Prevention Departments (PD). An introductory training to such programme was devised in the PD of the Local Health Unit no.11 Empoli, since the management was persuaded that all staff should be provided with basic knowledge concerning quality before being enrolled in QM programmes. Managers in charge of PD Units were offered a one-day training,focused on their role in promoting and achieving a QM culture, as an opportunity to be directly involved in the drafting of the DP quality handbook and in improving working procedures. The whole staff took part in a more extensive two-days course, including the basic concepts about QM in health systems, and the illustration of the programme in Regione Toscana; they were also asked to draw up some working procedures according to the draft already used in the Local Health Unit no.11, to be included in the DP quality handbook. The goal of this training programme was to implement a process capable of improving integrated working procedures through the competence, autonomy and responsibility of PD professionals. PMID- 18409763 TI - [An integrated prevention intervention for the management of SSUEm118 operator with hearing loss]. AB - Noise risk is moderate in hospital but there are some areas, as SSUEm118, where the background noise is not negligible. In this context it is important not to underestimate hearing loss even minimal, because the noise interference can cause errors of understanding, as the case in exam: an operator 118. The clinical assessment of this worker with liminar tonal audiometry and vocal audiometry revealed a mild unilateral mixed hearing loss with good voice response. The equivalent sound pressure level (Leq) throughout the SSUEm118 area is under the first attention level, the index NR is comparable to office type environments and the distance between speaker and listener for satisfactory face-to-face communication is suitable. The present study proposes a multi-multidisciplinary approach to define prevention and protection measures and health surveillance plan with audiometric testing and possible judgment of the unfitness to specific work. For this specific case advices given to SSUEm118 administrator allowed a good repositioning of the worker. PMID- 18409764 TI - [Rhabdomyolysis in a worker exposed to paraffinic mineral oils. Unusual association with bronchiolitis obliterans and organizing pneumonia (BOOP)]. AB - We describe a 29-year-old worker; exposed to metal dust, sawdust and paraffinic mineral oils in a factory producing accessories for leather articles and clothing, who came to observation with fever (39 degrees C), chest pain and marked increase of muscular enzymes. Chest computed tomography showed two areas of lung consolidation, that subsequently appeared enlarged with central escavation. Lung biopsy lead to the diagnosis of BOOP. The case has been reported to the judicial and workers' compensation authorities as probable occupational toxic myopathy. Indeed, exposure to paraffinic mineral oils is a known, though unusual, cause of rhabdomyolysis. The association with BOOP has never been previously described. PMID- 18409765 TI - [Occupational exposure and chronic heart failure severity]. AB - Chronic heart failure (CHF) is characterized by the inability of the heart to supply the body with sufficient amount of blood for metabolic and circulatory needs. The main risk factors for CHF development are: hypertension, type 2 diabetes, obesity, smoking, chronic kidney diseases. Many occupational exposures, such as extremes of heat or cold temperatures, prolonged exposure to noise, vibrations, pesticides, can contribute to etiology of this disease. The aim of our study was to evaluate if work can affect CHF severity. We analyzed retrospectively the first 76 smokers aged over 65 years who presented to the outpatient Clinic of Chronic Heart Failure. The patients were divided in 4 groups based on their previous job: white-collars, farmers, steelworkers and subjects performing different occupational activities (hairdressers, firemen, masons). Our results showed that farmers had a reduced left ventricular ejection fraction compared with white-collars (p = 0.0045) although NYHA class and the presence/absence of CHF risk factors were not different between the two groups. This data suggests that the farmer job could be associated with the severity of CHF. PMID- 18409766 TI - [Scleroderma cluster among type-setters]. AB - The etiology of systemic sclerosis, probably multifactorial, is not yet well defined. Among the many endogenous and exogenous factors probably involved, occupational elements may play an essential role. Here we report a cluster of local scleroderma and systemic sclerosis, which occurred in a small group of typography workers exposed to polyvinyl-acetate glues, containing up to 1% of vinyl-acetate. Vinyl acetate exposure has been associated with acidification of the intracellular environment, which is thought to produce cytotoxic and/or mitogenic responses that are the sentinel pharmacodynamic steps toward cancer. Autoantibody production in systemic sclerosis depends upon intracellular acidification. More studies are needed to clarify the relationship between vinyl acetate exposure and scleroderma. PMID- 18409767 TI - [Exercise electrocardiography in the sanitary surveillance of workers with physical strain]. AB - To evaluate the opportunity of exercise electrocardiography (ECG) in the sanitary surveillance of workers with physical strain, we estimated the energy consumption of the duties of 22 electrical workers (males; age: 35-56 years). They subsequently underwent Treadmill exercise ECG, determining for each worker the maximal METs (multiples of basal metabolism) and the critical potency (P(CRIT)). In one subject, myocardial ischemia arose 9 minutes after the beginning of the test. The others interrupted the test after 7-13 minutes for tiring; 5 of them showed ventricular extra systoles, paired or isolated. Twelve subjects presented arterial hypertension, at rest and/or during exercise. Ergonomic analysis revealed that the occupational duties were between 1.5 and 8.0 METs. The energy consumption of the job on the whole was 4-6 METs (medium intensity). The maximal METs reached by the examined subjects were between 8.8 and 15.6; however, only 11 workers went reassuringly over the 4 METs required by duty analysis at P(CRIT). One subject was declared unfit for the job, and a judgement of partial idoneity was expressed for 3 workers. Preventive and therapeutic indications were given to 12 subjects. Aerobic training was suggested to 10 workers. The study indicates that an ergonomic evaluation is advisable for the most energy consuming occupational duties. In such cases, the sanitary surveillance should include a cardiologic assessment with exercise ECG, reproducing the physical strain of the specific job. PMID- 18409768 TI - [Specific immunotherapy and relocation in occupational allergic bakers]. AB - Occupational allergy to components of wheat flour is the main cause of rhinitis and asthma of workers in bakeries and similar activities. An immunological mechanism IgE-mediated is involved and the sensitising properties of some proteins of wheat where assessed. Nowadays it is possible to have an extract to be used for specific immunotherapy. The aim of this treatment should be a reduction of individual immunological reactivity and the possibility of going on the particular activity of allergic bakers, pastry makers or pizza makers. An observational crossectional retrospective study was performed on 41 sensitised workers that were diagnosed in the same occupational health unit. All underwent a subcutaneous specific immunotherapy (SCIT) with the same schedule and the same extract (Lofarma Allergeni, Milan) for 4 or more years, without avoiding their work activity. The outcome was investigated after five or ten years. Data were collected by a questionnaire. 34 subjects on 41 are still at work with an acceptable quality of life and a normal working efficiency, mainly in their small enterprises. In the "old" subgroup (19 cases), treated in the past, several bakers still at work stopped SCIT even from 4-10 years. In the "new" subgroup (15 cases), still in treatment, symptoms and drug use during the work activity resulted to be reduced or absent in the majority of cases. According to results of other immunotherapies by allergenic vaccines (pollens, mites) also for wheat flour occupational allergy a specific treatment seems to be possible and SCIT may be an useful tool to reduce and control the biological individual effects of allergy. By the occupational point of view wheat flour SCIT allows a relocation in many of cases and may be associated to other intervention of environmental prevention at workplaces, improving the relocation of occupational allergic subjects when requested. PMID- 18409769 TI - [Metabolic syndrome and work: identification of populations at risk]. AB - OBJECTIVE: To assess whether shift work has a cause-effect nexus with the incidence of metabolic syndrome. DESIGN: Retrospective follow-up study, case control type. RESEARCH METHODS AND PROCEDURES: A total of 202 female and male nurses, aged 23-60 years, having a 1 year minimal follow-up, and without any metabolic syndrome criterion at the first visit were evaluated. The ATP modified criteria were applied for the diagnosis of metabolic syndrome. Moreover smoking habitus, alcohol consumption, educational level and physical activity were considered as bias factors. The sample was divided in two groups doing or not night shifts. RESULTS: The metabolic syndrome incidence resulted significantly greater in shift workers than in other workers (OR: 4.10, 95% CI 1.34-12.55, p = 0.01). The difference was more evident in subjects aged < 40 years (OR = 6.6, 95% CI 1.05-40.85, p = 0.04) the different metabolic syndrome incidence between two groups was detectable even after the multivariate analyse, which considered all the bias factors evaluated. CONCLUSIONS: This study shows for the first time that the night shift work is associated with a greater risk to develop metabolic syndrome in workers healthy in baseline conditions. PMID- 18409770 TI - [Metabolic syndrome in visual display units users]. AB - BACKGROUND: Reports about medical consequences from sedentary work are contradictory. It might be associated with the metabolic syndrome (MS), a collection of cardiovascular risk factors including hypertension, dyslipidemia, insulin resistance and central obesity. No data are currently available on workers using visual display units (VDIU), a potential high risk group, given the sedentariness inherent in this work. PATIENTS AND METHODS: We evaluated MS prevalence in 1547 VDU users with a mean age of 29.7 years and in a control group of 892 individuals with a mean age of 30.2 years who performed non-sedentary work, selected on the basis of similar demographic data. Physical examination and laboratory tests useful for MS diagnosis were performed. RESULTS: MS prevalence was 3.10% in VDU users vs 2.01% in controls (OR 2.048, 95% CI 1.169 to 3.587, p = 0.012). Significance persisted after controlling for confounding factors (e.g, smoking and leisure activity) in a multivariate analysis (OR 1.555, 95% CI 1.03 to 2.690, p < 0.05). CONCLUSIONS: MS should carefully considered when performing health surveillance programmes in VDU users. PMID- 18409771 TI - [The reference values selection and validity check also are a spirometry quality issue]. AB - Spirometric measurements are widely used for routine clinical assessment of workers with occupational exposures to bronchial irritants or sensibilizing agents, however their interpretation requires appropriate reference values for predicting normality. We studied 1319 subjects 18-25 years old (754 males--57.1% e 565 females--42.9%) with BAIRES-Biomedin spirometer and observed values were compared with different reference equations for italian workers: CECA 1971, ERS 1993, Scotti and coll. 1986, Pistelli and coll. 2000. All reference equations studied have significantly differences for one or more index, even if the observed values are all near to 100% of predicted values. The ERS 1993 reference values were always statistically different from observed values. In conclusion we suggest prudence in interpreting spirometry in young people and we hope the identification of new spirometry reference values for Italian people. PMID- 18409772 TI - [In vitro percutaneous absorption of silver nanoparticles]. AB - There is a growing interest in the debate on nanoparticle safety for topical use. The benefits of nanoparticles have been shown in several scientific fields, but little is known about their potential to penetrate the skin lies. This study aims at evaluating in vitro silver nanoparticles skin penetration. Experiments were performed using the Franz diffusion cell method with intact and damaged human skin. Physiological solution was used as receiving phase and 70 microg/cm2 of silver nanoparticles dispersed in synthetic sweat were applied as donor phase to the outer surface of the skin for 24h. The receptor fluid measurements were performed by Electro Thermal Atomic Absorption Spectroscopy (ETAAS). Silver concentration of 0.2 microg/L was found in the receiving solutions of two cells, in which damaged skin membranes were set up. In the other tests, we obtained a silver concentration below the limit of detection in the receiving cells. Our experimental data show that silver nanoparticles permeation through intact and damaged skin is negligible. These findings are consistent with previously published results. Further researches are necessary to explore skin absorption of silver nanoparticles. PMID- 18409774 TI - [Dermatitis and VDU work]. AB - Skin disorders like rosacea, seborrhoeic dermatitis, non-specific erythema and acne can be VDU work linked. At present, many environmental and individual causes are involved in the development of these disorders, but the former appear to be the issue to investigate more in depth. PMID- 18409773 TI - [In vitro percutaneous absorption of chromium powder and the effect of skin cleanser]. AB - Occupational chromium dermatitis occurs frequently among cement and metal workers, workers dealing with leather tanning and employees in the ceramic industry. The present study, using an in-vitro system, evaluated percutaneous absorption of chromium powder and the effect of rapid skin decontamination with a common detergent. Experiments were performed using the Franz diffusion cell method with human skin. Physiological solution was used as receiving phase and a suspension of chromium powder in synthetic sweat was used as donor phase. The tests were performed without or with decontamination using the cleanser 30 minutes after the start of exposure. The amount of chromium permeated through the skin was analysed by Inductively Coupled Plasma Atomic Emission Spectroscopy and Electro Thermal Atomic Absorption Spectroscopy. Speciation analysis and measurements of chromium skin content were also performed. We calculated a permeation flux of 0.843 +/- 0.25 ng cm(-2) h(-1) and a lag time of 1.1 +/- 0.7 h. The cleaning procedure significantly increased chromium skin content, whereas skin passage was not increased. These results showed that chromium powder can pass through the skin and that skin decontamination did not decrease skin absorption. Therefore, it is necessary to prevent skin contamination when using toxic agents. PMID- 18409775 TI - [Night workers and plasmatic cortisol]. AB - The aim of the study is to evaluate whether occupational exposure to night work could cause alterations in the levels of plasmatic cortisol. The interest toward this argument arises form several studies in scientific literature referring the presence of an alteration in the synthesis and release of cortisol in workers exposed to night work. We studied a population of workers employed in night security service and monitoring service of alarm systems in different museums compared to a control group not performing shift-work and/or night work. The exposed and control subjects were compared by age, length of service, smoking habit (n. cigarettes per day), habitual consumption of alcoholic drinks (n. glass of wine/beer per day). We evaluated the levels of plasmatic cortisol on 50 workers exposed to night work, all males of whom 30 smokers and 20 non-smokers and on 50 controls of whom 30 smokers and 20 non-smokers. PMID- 18409776 TI - [Effectiveness studies in the Italian literature on occupational health (1990 2006)]. AB - INTRODUCTION: Evidence-based prevention (EBP) is gaining the growing interest of the scientific community in Occupational Health (OH). To highlight the Italian contribution to EBP, a systematic review was performed, on the main OH-journals publishing in Italian language. METHODS: Studies that have evaluated interventions carried out to improve workers' health were found out from the articles published in the period 1990-2006, by means of hand-searching. The selected articles were classified on occupational health topics as health intervention, study design, risk factor, authors affiliation. RESULTS: 16 publications were selected, which represent less than 0.5% of all papers published on the 8 Italian journals examined. The most frequent study-design is the pre-post intervention evaluation; 2 out of 16 (25%) studies are randomized trial. The interventions are directed mainly to control the effects of noise and biomechanical overload; textile, health-care and energy production are the most frequent industrial settings. We did not found researches aimed to evaluate the effectiveness of interventions performed in the metalware industry or addressed to injuries prevention. DISCUSSION: The systematic approach of EBP can be applied to most of the occupational health interventions and effectiveness evaluation should represent a challenge for the occupational health field. The results of the hand searching among publication in Italian could raise interesting debate about methodology and outcomes in the scientific community. PMID- 18409777 TI - [Study of the olfactory function of a group of workers with significant lead exposure]. AB - The aims of this study are to verify the potential lead damage on olfactory function and to identify early effects due to lead exposure. Our diagnostic evaluation included: (i) questionnaire to collect data about work and clinical history, (ii) olfactory evaluation: threshold test (Single-Starcaise) and identification/discrimination test (Wright). Lead exposure was evaluated by air sampling and biological monitoring (PbB, lead in blood). A sample of 18 exposed workers (mean age: 41.3 +/- 7.8; years exposure: 8.38 +/- 6) and of 39 controls (mean age: 41.9 +/- 9.7) were evaluated. The comparison between the threshold test of two groups confirmed a worse olfactory function in exposed (-4.97 log(10)vol/vol) compared to controls (-6.37 log(10)vol/vol), while the Wright test didn't show any significant correlation. The study didn't find a significant association between individual PbB levels and the threshold test. Knowledge of the effect of chronic occupational exposure to industrial chemicals on olfactory function is largely incomplete, but supports the hypothesis that olfactory neuroepithelium is susceptible to environmental exposures to chemicals. Occupational-related olfactory impairment is usually sub-clinical, and can be only detected using adeguate quantitative olfactory function testing procedures for quality research in this field. PMID- 18409778 TI - [Nanoparticles and health effects: an evidence based approach]. AB - The possibility of health effects related to nanoparticles (less than 100 nanometers diameter) exposure may be considered as an emerging problem. Current approaches are usually partial and do not consider the multidisciplinary available data. The purpose of this work is to define the problem using an evidence-based approach: we characterized the possible nanoparticles-human interactions by collection of all pertinent scientific data available in the literature and by their critical evaluation. The distinction between non intentional nanoparticles ("ultrafine particles", UFP) and intentional nanoparticles ("intentionally produced nanomaterials" or simply "nanomaterials", IPN) is a critical item. Each single type of nanoparticle is more relevant than isolated metric characteristics. Globally, the experimental studies suggest a possible inflammatory acute effect. The available data regard mainly the UFP. Regarding IPN, the studies about attaints on human health are actually few and limited: no cardiovascular. respiratory and coagulation effect were showed as consistent with exposure. No human study about IPN chronic effect is available, and any human data is not showed relating carcinogenic and sensitizing effects. PMID- 18409779 TI - [Interaction between carbon nanotubes and cardiovascular autonomic nervous system regulation: proposal of an animal model and preliminary findings]. AB - BACKGROUND: Altered autonomic cardiovascular regulation (ACR) may mediate the association between single-wall carbon nanotubes (SWCNT) exposure and adverse cardiovascular events. MATERIAL AND METHODS: 400 mg of SWCNT in 400 ml of phosphate buffer saline (PBS) or 400 ml of PBS were randomly given to 7 Wystar Kyoto rats (400 g body wt) previously implanted in abdominal aorta with a telemetry transmitter for recordings of arterial pressure signals. Recordings were performed at baseline, 24 hours and two weeks after intratracheal instillation. The beat-by-beat time series of systolic arterial pressure (SAP) and PR interval were analyzed to identify sequences of three or more consecutive beats in which SAP and PR changed in the same (baroreflex sequences) or in the opposite direction (nonbaroreflex sequences). The mean individual slope of the sequences was calculated and taken as a measure of the baroreflex (BRS) and nonbaroreflex sensitivity for that period. RESULTS: The 24 hour BRS response showed a 100% increase (from 4.6 to 9.2 msec/mmHg) in controls, whereas it was blunted in cases (from 5.1 to 6.1 msec/mmHg) (p < 0.05). CONCLUSIONS: Our study demonstrates that this rat model is suitable to study the ACR during exposure to SWCNT and suggests a blunted BRS response after SWCNT instillation. PMID- 18409780 TI - [Compliance to regular health surveillance in workers with different work related risk factors]. AB - The Author examined in The Time of Twelve Months (October 2006/October 2007) a sample of 371 Workers exposed at different Types of Risck Factors (See Tab. I) with the Aim of to Express one Judgment of Fitness at The Specific Task and PreVenction and Therapie of the Professionals Diseases. The 74.52% of the Workers to Obtain one Judgment of Fitness Absolute and the 25.48% one Judgment of Fitness with Specific Limitation. The Casistic to understanding a Number of 316 Male (85.17%) and 55 Female (14. 83%). The Middle Age of the Sample it's 38. 17 Year with DS:9.77, and it's therefore juvenile, middle. To come Explained in the Tab. III the Percentage of Internistic Diseases obtained in The Fitness with Limitation. The Symptoms of Phatologie most attend aren't: The Arterial Hypertension (29.17%), following to Low Back Pain/Artrhosis (15.63%) afterwards the Ear Deafness for Noise (14.58%) and The sharp or Chronic Bronchitis (12.50%). The Subject Workers aren't dependent to Questionnnaire about the Job Satisfaction (JCQ di Karasek R) about the Stress (Pisa Stress Questionnaire of CA Pruneti at 32 Items) and about the Alexithimic Characteristic (Test Tas-20). The Outcomes aren't explained in Tab. IV. It's Already explain, that Judments of Not Fitness absoluted to have Heart repercussion about the Psychosomatic Health of the Workers and of Your Family System with one increased Psychosocial Risck and Inability, and therefore Judgment of Fitness at Work with specific Limitation aren't to indicate for adaptation the Worker at the best Work and Task. PMID- 18409781 TI - [Passive smoking at work: proposal for a smokeless company policy]. AB - Tobacco smoke contains about 4000 chemical substances, among which cancer agents (benzene, formaldehyde, polonium-210). There is sufficient evidence that inhalation of tobacco passive smoke causes human risks (cancer, cardio-vascular and respiratory system diseases). With the no-smoking act, issued by minister Sirchia in 2003, smoke is forbidden in indoor space in Italy, but much has still to be done in practice. Difficulties in application depend on the facts that smokers psychologically and hardly oppose and sanctions are complicated to apply. In this work we will argue the following problems and purpose respective alternatives: 1) To consider passive smoking risk in the 626/94 act into a specific section. 2) To transform administrative sanctions into a fund finalized to smoke cessation programs. 3) Increase formation-information smoke cessation programs, especially focused on positive effects from stop smoking. PMID- 18409782 TI - [Biostatistical study on health surveillance of Military Navy personnel in Sicilia and Calabria regions]. AB - The Maritime Military Health submit to periodic medical examination civilian employees exposed to health risks and all military personnel. The latest, because of particular use in operational duties, as well as the characteristic of seafarers employed, undergoes an extensive medical protocol. In 2006 there were around 3200 access to the Service of Medicine Labor Navy Hospital in Augusta (SR). We wanted then to make a collection of data with purpose to highlight the diseases that most often affect the area of military jurisdiction. The illnesses were divided into two main groups: those who did not produce any result on the job because of insignificant impact functional, and those for which they have been taken to unfitness, temporary or permanent. From a statistical evaluation of the acquired data, it results that most affected systems are the skeletical, hematologic, psychic, ENT, heart, the Body Mass Index. PMID- 18409783 TI - [Fitness to work and diabetes in health care workers]. AB - Diabetes mellitus is a disease of great social relief that has a deep impact also in working life. Our short paper wants to highlight some critical aspects that occupational physician must deal with both in preventive and periodic health evaluation for diabetic subjects. In our study, performed on health care workers, we reported 16 cases of diabetes; 10 of them were nurses, 5 sanitary auxiliaries and 1 sanitary technical operator. The judgment has been of full suitability to the specific task for 7 subjects, whereas for 4 workers it has been stated suitability with limitation for problems related with diabetes or its complications. The judgment with limitation for the other 5 subjects was due to pathologies not related with diabetes. The shift work, and mainly the night shift, is the risk that diabetic subjects cannot effort when the disease is not sufficiently compensated. The small number of diabetic workers, 16 over 1994 visited (0.8%), suggests for a spontaneous choice of diabetic people towards other kind of work without shift risk. PMID- 18409784 TI - [Health surveillance in ridden work environments by means of the MM040/IAQ Questionnaire]. AB - Complaints and symptoms related to the indoor environment experienced by 3380 workers were collected from 26 workplaces by using the Italian modified version of the Swedish questionnaire MM-040 and the Finnish IAQ-Indoor Air Quality questionnaire. Complaints related to environmental factors, and air related symptoms, were common in indoor workers. Women reported work related symptoms more often than men, the difference being significant. The Indoor Air Quality questionnaire is a suitable tool for the occupational health physician in investigating indoor air problems. In the Italian version six general symptoms were added to the symptom scale, in order to make the questionnaire feasible for analysis of general health, during medical surveillance at work. PMID- 18409786 TI - [PC use in university students]. AB - This study was aimed at investigating features related to PC use and prevalence of musculoskeletal symptoms among university students. A cross-sectional study was carried out among 98 university students. A questionnaire was used to collect data on personal characteristics, PC use and symptoms prevalence. Mean duration of daily PC use was 8,3 (DS 1,9) hours. The prevalences of neck, hand/wrist and shoulder pain in the previous month were 76%, 69% and 56% respectively. The findings of the present study suggest the needing of specific ergonomic training in university students. PMID- 18409785 TI - [The physician with a reduced work ability]. AB - The occupational health physician charged of medical surveillance of hospital workers is often mandated to manage impaired physicians. Some "red flags" may be observed during medical surveillance of workers, and this should elicit the suspect that the colleague is impaired by chemical, psychiatric, or neurological disease. Three emblematic cases are discussed. While English-speaking countries have faced the problem since the 70's, Italy currently lacks policy to deal with impaired physicians. PMID- 18409787 TI - [Application and effects of smoking ban in bars and restaurants of Rome]. AB - Both active and passive tobacco smoke is carcinogenic. In the last years the most important countries of European Community developed and passed smoke-free public places and smoke-free workplace legislations. The aim of this study was to investigate the real application of smoking ban in bars and restaurants of Rome and to value social, economic and health effects caused by the application of the law. The study was carried out in 200 public places (100 restaurants and 100 bars) with an inspection of the sites and the administration of a questionnaire to the managers of the public places. Results demonstrate that smoking ban in public places is widely respected and that the application of the law had a very positive impact on the quality of life and health of workers and general population. PMID- 18409788 TI - [Institute for preventive medicine and health promotion for a population based on university students]. AB - During academic year 2006-2007 we have been trying to restart the Institute for Preventive Medicine of Universita Cattolica in Milan for all university students. Thanks to a co-operation between Milan Universita Cattolica Health Centre and the Postgraduate School in Medicine for Work of Universita Cattolica of Rome, we have studied with the utmost care the structural and functional background of the Health Centre. Through questionnaires we have calculated all students' needs and if they felt quality in the service. In addition to that, we moved on and we investigated all ideas relating to future projects in order to restart all Institutes for Preventive Medicine in each University without additional costs for all University administrations. PMID- 18409789 TI - [The demonstration of the connection of cause in the professional allergic pathology not tabulated. Utility and limits of the allergometric tests]. AB - The Sentences of the Constitutional Court no 179/88 and 206/88 have of done modified the system insurance of the professional illnesses. To the old tabular system to the pathologies for which the legal conceitedness of origin is in force they are added those pathologies that need to be shown with burden of test of the applicant. And necessary to resort to the criterion pathogenetic for the geneses pathologies plurifactorialy or to non certain etiology. These innovations determine the necessity of a methodological iter forced for the recognition of the professional illness not table. The authors therefore they propose an iter diagnostic that passes through the recognition of the connection of cause. To such intention the authors examine meaning and limits of the suggested investigations. PMID- 18409790 TI - [Body mass index in shift workers: relation to diet and physical activity]. AB - Many studies investigated adverse effects of rotating shift work, deriving from modifications of circadian rhythms and alimentary behaviours. This may cause the disregulation of glycolipidic metabolism and of insulin secretion, and the development of central obesity. We conducted the present study in order to assess the differences in body mass index (BMI) between day and shift workers, in relation to diet quality and physical activity. DESIGN AND METHODS: The study included 341 male subjects (165 day workers and 176 shift workers). Body weight, height, BMI, waist and hip circumferences were measured. Physical activity and diet quality were assessed by self-reported questionnaires. RESULTS: Shift workers had higher BMI than day workers (27.6 +/- 3.9 vs 26.7 +/- 3.6), and this correlation persisted after adjustment for age. The diet quality was better in shift workers and the level of physical activity was similar in the two groups. DISCUSSION: The results of this study indicate that shift work is an independent risk factors in the development of overweight and central obesity. In fact, although the diet quality was even better in shift workers, and the level of physical activity was similar between day and shift workers, we found that shift workers had higher BMI. PMID- 18409791 TI - [New tools for the study of occupational allergies related to animals]. AB - Animal laboratory workers are exposed at the Laboratory Animal Allergy (LAA). About 10% of them develop asthma. Currently, by the use of innovative methodologies, numerous allergens from the laboratory animals have been sequenced and analyzed. Between them, the allergens by rats and mouse, the experimental models more utilized. Proteomic approach or protein microarray permit the study of several allergens, belonging to the proteins known as lipocalins, and of immunological response in susceptible individuals. Moreover, availability of on line data banks permit a knowledge more and more detailed and up-to-date regarding the allergens of the interest. Between the advantage of the proteomic there is the use of small amount of biological fluids, as the serum, in association with the possibility to studied a great number of allergens on a single support. The study of LAA by the use of proteomic will permit to identify the animal allergens more accurately, providing also a valid tool at the methodologies traditionally used, and for the planning of the preventive and protective measures. PMID- 18409792 TI - [Tobacco smoking among Italian physicians]. AB - A survey of the international literature published between 1985 and 2000 reveals high smoking rates among Italian physicians. Since 1985 smoking rates among physicians have gradually declined, similarly to those of general population. However, prevalence rates seem to vary between northern and southern Italy, with the highest rates in the southern regions. Studies examining smoking rates by gender reveal that, while smoking trends among males have declined somewhat, the latter remained relatively stable among female physicians. From an international perspective, declines in the absolute smoking rates of Italian physicians have not kept pace with those of many other countries, since at least one-quarter of Italian physicians currently smoke. Furthermore, most of the Italian physicians smoke while they are at work. This represents an occupational health problem that needs to be addressed by all levels of management. In meeting this challenge, occupational medicine has an important role to play in helping reduce the prevalence of smoking among health care workers. PMID- 18409793 TI - [Tobacco's smoke in the workplaces, webpages in the ISPESL's website]. AB - The authors present the section, part of the ISPESL's website, dedicated to tobacco smoke at work. In this subdivision many topics regarding problems caused by tobacco smoke in the workplaces are gathered and discussed so that different personnel responsible for health and prevention at work can find a technical answer to take part to the improvement of the psychophysical welfare of both smokers and non smokers. The general information section has collected the relative Italian and international laws regarding smoking in the workplace along with representative court cases, and some publications and essays which have been presented on this topic. Inside the section dedicated to the employers and companies, the authors have posted useful importation on smoking in the workplace which includes some tools that can turn to be interesting to those who are supposed to be involved with prevention, the updated addresses of the national public antismoking centers, experiences and activities against tobacco smoke of national and international companies, events regarding smoke at work, informative sheets and leaflets for smokers, link to other site that argue about this topic. Moreover, there is a section dedicated to a forum where Internet user can share their experiences and thoughts on workplaces free from tobacco smoke. PMID- 18409794 TI - [Job fitness management regarding university school of nursings students: our experience over three years (2005-2007)]. AB - Based on definition of worker according to D.Lgs.626/94, art.2, c.1 also the University Students before to start their pratical training in Hospital must be submitted to sanitary surveillance by Occupational Health Physician. Aim of this paper is to report about the Job Fitness Management regarding no. 231 University School of Nursing's Students that in some cases has involved the formulation of limitations and/or prescription. PMID- 18409795 TI - [Pilot study of professional exposure and hearing functionality of orchestra musicians of a national lyric theatre]. AB - The sound exposure of classic orchestra musicians is not well assessed in Italy even though the protectionistic law includes all the working environments, as stated in the European Directive 2003/10/EC relative to the protection of workers against noise-induced damage. Scientific data in the literature show that the musicians are exposed to high daily personal exposure levels. These levels can systematically exceed the 85 dB(A) in the case of percussionists. In this study, preliminary results are shown relative to an experimental campaign performed in a lyric National Theatre. The aim of the study was the assessment of the risk due to high sound level in the theatre musicians and their cochlear functionality. The noise levels were measured by using wearable phonometers and multichannel sound analyzers. The signals were recorded during the whole working time in the rehearsal hall and in the fossa. The cochlear functionality was monitored before and immediately after rehearsal in a sample of volunteers by means of otoacoustic emission tests. Both TEOAE and DPOAE were measured using the ILO292 portable system. The data show that OAE-based tests can effectively discriminate between different classes of exposure. A good correlation was found between the daily exposure level and the DPOAE level. PMID- 18409796 TI - [Preventing measures facilitate the preservation of the levels of employment]. AB - The implementation of the European Directive No 391/89, among the other news, introduced the obligation for the occupational physician to actively participate to the assessment of risks. The reduction of the incidence of "historical" professional pathologies and the arising of work-related disease is making the role of the occupational physician, in the survey and risk assessment, ever more important, especially in consideration of the physiological and biomechanical peculiar competences required. In addition, a correct risk assessment allows to optimise the valorisation of human resources. The occupational physician, in other terms, can not only protect the health of workers, but "save" jobs too. The survey carried out in a logistic unit of a large-scale distributors group, documented herein, can give evidence of that. PMID- 18409797 TI - [Asthenopia in VDT users: our experience]. AB - The aim of this study was to investigate the relationship between asthenopia and work with video display terminals (VDT). 62 office workers (29 males and 33 females), that use VDT for more than 20 hours each week, were investigated by means of standardized questionnaires (elaborated by the Italian Society of Occupational Medicine and Industrial Hygiene). The authors analyzed the prevalence of occupational asthenopia and its correlation with the duration of weekly VDT use and the presence of refractive alterations. It was demonstrated a prevalence of asthenopia of 51% and a statistically significant correlation between the entity of asthenopic symptoms and number of weekly hours and years of work at VDT, in particular in subjects with presbyopia. PMID- 18409798 TI - [Smoke habit in a population of apprentices in middle of Italy]. AB - This study assessed the smoking habit in 7,159 apprentices (females: 53.3%) in central Italy. In the period 1996-2006, each apprentice underwent a compulsory health examination to assess fitness for work carried out by specialists in Occupational Medicine. Demographic data and information on schooling, smoking habit, alcohol and coffee intake were collected by a questionnaire. Overall, 51.9% of apprentices were smokers. Smokers were fewer among female apprentices than among the males (48.3% vs 55.9%; chi-square test, p < 0.001). No decrease in the prevalence of smokers was evident during the study period. Low educational level, and daily intake of alcohol and coffee significantly increased the risk of being a smoker. Finding a high percentage of smokers among Italian apprentices with a low educational level who are employed in jobs for which few qualifications are required indicates the need for work-place activities against tobacco smoking habit. Specialists in Occupational Medicine who perform regular health surveillance of workers could play a role in the anti-smoking campaign by integrating their routine work with brief counselling sessions aimed at preventing smoking in young workers and helping smokers to stop and by collaborating in work-place health promotion programmes. PMID- 18409799 TI - [Smoking habit among workers in Campania region]. AB - Smoking is still now the main avoidable cause of disease, disability and mortality in industrialized countries. This habit is still very common in workplaces, where anti-smoke efforts seem to be less incisive than among general populations. The study analyzed the diffusion of smoke habit in 8111 male workers in Campania region, employed in different work activities (white collars, blue collars, drivers, cleaning civil servants, porters), so as to evaluate work related features, affecting its assumption and maintenance. Among all workers, smokers prevalence (42.7%) was higher than national male population. Percentages of smokers were highest among drivers (60.7%) and civil servants (52.5%), slightly lower among industry workers (47.3%) and lower among white collars (36.4%). The highest prevalence were found in 41-50 years age group (46.8%), but only among white collars aging was associated with higher smokers prevalence. Lower education degrees and two working variables, shifts and handwork, have been related with significantly smoking habit assumption. Results emphasized that health promotional programs are necessary to reduce smoke habit among workers, particularly among professionally and culturally unqualified subgroups. PMID- 18409800 TI - [Suitability to the specific task of drivers suffering from hypertension]. AB - The italian legislation defines the minimal psychophysical requirements for driving licence, that is indispensabile to be employed, but leaves extensive discretion in the evaluation of the cardiovascular diseases. Hypertension, the most important risk factor for heart disease, is not considered a contraindication to drive. Uncontrolled blood pressure or target organ damage expose to acute complications that may well preclude safe driving and be the cause of crashs; therefore the role of the occupational physician is so important in the sanitary surveillance of the drivers of vehicles for transportation of people and/or goods. PMID- 18409801 TI - [Formation and quality: an integrated pattern of professional growing]. AB - The cultural formation the professional man's knowledges and his way to operate. It represents a resource to elaborate organizational and operative methods compatible with the user's needs. A good formation permeates the professional man's processing technique and it accompanies the definition of strategies, targets and decisions. The formation makes the professional man acquire specialist knowledges and actions to reduce the fragmentation of the human communication and it favours the efficacy of all the interventions for the health and the security of employment. It is necessary to promote many operative methods integrated with a qualitative system in order to favour the managerial abilities of a professional man together with his competences inside our social and productive world. PMID- 18409802 TI - [Alcohol levels determinations in workers communities: a standardised practice proposal]. AB - Italian law 125/2001 implementation requires to Occupational Health Physicians to carry out alcohol levels checks in the workplaces. Firstly, this contribution proposes to specify the methodological differences between breathing alcohol concentration, BrAC, and blood alcohol concentration, BAC, and also to show the different sampling strategies. Therefore, a standardised practice proposal for alcohol levels determinations in workers communities is presented, in the contest of a modern Occupational Health services "integrated quality system". PMID- 18409803 TI - [Work and alcohol abuse: the issues]. AB - Law n. 125 "Framework law regarding alcohol and alcohol related problems" came into force in Italy in 2001. According to WHO, approximately 10-30% of work accidents would be alcohol related. Alcoholic beverages at workplace may produce an even remarkable increase of direct risks, because of psychophysical alteration and indirect risks produced by added effect of alcoholic beverages with industrial toxic substances. Even with low alcohol rates, the reaction time becomes longer, error frequency in response to visual or hearing stimulations is higher and hence the risk of accidents becomes sensible. Besides, alcohol-related working performance problems may be caused not only by drinking at work but also by drinking a lot of alcoholic beverages outside working time. In particular, art.15 of L 125/01 in compliance with alcohol use at workplace, forbids consumption or administration of alcoholic and superalcoholic beverages during activities at high risk for safety. In this context, the norm provides power-duty to make alcoholmetric concentration checks only to company physician and prevention and safety occupational health service of national health units. PMID- 18409804 TI - [Development of an educational programme for schools to promote the awareness on child labour]. AB - The main aim of the ISPESL project "Child Labour: to know in order to take action" is to introduce the subject of the problem of child labour in primary schools. This is done through pedagogical tools and communication methods, using the child's curiosity and point of view as a starting point and giving support to the child as he/she expresses his feelings, messages and emotions and at the same time, develops his own reaction to the problem. This project has been developed within the activities of ISPESL as WHO Collaborating Center and addressed to all the reference figures interacting with school such as teachers, educators and headmaster, but also families and local Institutions who promote understanding and awareness on child labour. The project aims at giving a general picture of the phenomenon, by focusing on the multiple and complex causes affecting the physical and psychosocial well-being of children and on fostering the increase of awareness among children on child labour issues. Observation and investigation tools will be developed and used in specific training modules suited to the cultural and geographic framework of each field of teaching, in order to allow the children to acknowledge and express their better awareness on child labour. PMID- 18409805 TI - [Risk factors for smoking among young apprentices]. PMID- 18409806 TI - [Consequences of work environment on the ability judgment in a hospital company in Piemonte]. AB - The aim of this study is to analyze whether the changes recently occurred in the Italian society and in the occupational context can influence the prevalence of job restrictions among health care workers (HCW) of the Cuneo Hospital, between 1998 and 2006. After subdivision in: nurses, physicians, auxiliary nurses, administrative staff, and technicians, sociodemographic data of HCW were collected. In 2006, the total amount (+176), number of part-time (+274), mean age (+2.6 ys) and seniority (+2.1 ys) of HCW--this latter not for physicians and technicians-- were significantly increased (p < 0.001) compared to 2000. The prevalence of job inability--obtained dividing all the new restrictions expressed for each year between 1998-2000, 2001-2003, and 2004-2006 by the number of HCW working at the end of each period--increased, respectively, from 12.1% to 12.7%, to 16.1% (chi2 = 18.6; p < 0.001). The higher percentage of inability was observed among physicians (+ 183%) and administrative staff (+7/1%). In each period, subjects with job restrictions resulted to be younger than the whole HCW (p < 0.001). In conclusion, the number of job restrictions is rising but it doesn't seem to be associated to the increase of age and seniority among HCW. PMID- 18409807 TI - [Evaluation of the working load in subjects using equipment fitted with VDU]. AB - In Italy the health and safety of VDU workers is regulated by section VI of Legislative Decree 626/94 (with later modifications). In compliance with this law, employers shall be obliged to analyse workstations, assess and reduce risks; they shall also identify any worker who habitually uses display screen equipment as significant part of his normal work (20 hours per week). Our study presents a new method, named WODSE (Workers of Display Set Equipment), for the objective evaluation of VDU workers effort. It is a software created for measurement of VDT usage time, able to calculate weekly use for each worker. We applied this method to tellers of a great national services agency, obtaining an improvement of VDU risk assessment and, consequently, management. PMID- 18409808 TI - [Health surveillance in health care workers]. AB - In a complex reality as the hospital, occupational physician plays a key role in risk prevention through health surveillance. The medical examination is intended to highlight any possible deterioration of workers' health and to detect workers' hypersusceptibility to occupational hazards. We report biostatistic data resulted from health surveillance conducted on health care workers in 2005 and 2006 in a universitary hospital, with particular regard to the judgement of the fitness to work and the reasons that has determined it. Our report, in agreement with data available in literature, shows that manual patient lifting is one of the most common professional hazards within the hospital and occupational physician must pay a special attention to it, promoting an integrated answer. PMID- 18409809 TI - [Feasibility study of biological monitoring of chemical agents by means of evaluation of the effects on their in vitro effects on the complement system]. AB - Immunological methods for the study of the plasma complement system have been standardized in order to be good and reproducible indicators of some biological effects of the substances under study in in vitro experiments. The substances tested were not capable of interfering within 10 times the possible hypothetical plasma concentration reached in vivo with the function of the different reagents used in the study of complement. Five substances (Skin-ACGIH) have been studied for their effects on the complement system in vitro; four of them could be fully studied (allylic alcohol, cyclohexanone, phenol, dimethylacetamide). After this deep insight we can conclude that: 1. These substances are capable of interfering with the immune response through their complement activating capacity 2. These substances, throughout complement activation, can induce inflammation and reduction of important defensive functions that are complement mediated. 3. The results obtained encourage to study the complement system and especially CH50 in workers exposed to the selected substances in order to verify the possibility to enclose this test in the medical surveillance program. PMID- 18409810 TI - [Isotopic dilution HPLC/MS/MS urinary trans, trans-muconic quantitative analysis among refinery workers]. AB - The determination of trans, trans muconic acid (t,t-MA) in human urine, a biomarker suggested by the American Conference of Governmental Industrial Hygienists for occupational benzene exposure, with a limit value of 500 microg/g creatinine, is usually performed by means of gas or liquid chromatographic methods. Almost all the HPLC methods make use of strong ionic exchange cartridges for sample cleaning, reverse phase separation and detection either by UV at lambda = 259 nm or; more recently, by electrospray tandem mass spectrometry: yet, not all of these methods have been validated for quantitative analysis considering also the matrix effect. This paper presents the quantitative analysis of t,t-MA in 94 end-shift urine samples from workers of an oil refinery performed by means of an HPLC/MS/MS analytical method that uses a commercially available deuterium labeled isotope as internal standard, that during the validation has highlited the problem of interferences due to urine ion suppression effect and to the interference from isobaric ions both for the analyte and the internal standared. The following mean values have been obtained: 47.37 microg/g creatinine for non smokers non occupationally exposed to benzene, 97.40 microg/g creatinine for non smokers exposed to benzene, 142.38 microg/g creatinine for smokers non occupationally exposed and 149.08 microg/g creatinine for smokers occupationally exposed to benzene. The results obtained demonstrate that using this analytical method for urinary t,t,MA it is possible to discriminate among groups with different levels of benzene exposure, due to all the possible benzenene sources: environmetal, occupational, due to smoking, and their possible combinations. PMID- 18409811 TI - [Study on the interaction between the skin detoxifying enzyme glutathione S transferase and the substances listed in the CE/39/2000 rules with the "skin" annotation, finalized to the biological monitoring of exposed subjects]. AB - The interaction among chemicals listed in the Directive CE/39/2000 with skin notation and glutathione S-transferase (GSTP1-1) was studied by following two different experimental approaches. The compounds were incubated with the purified GST isoenzyme GSTP1-1 as well as with the human keratinocytes (PR5) selectively expressing GSTP1-1. Some of the molecules affected the enzymatic activity of both the purified and the intracellular GSTP1-1. In particular, 1,2-dichlorobenzene (DCB), ethylbenzene (ETB), cumene, Sulphotep and 2-eptanone (2-EPT) behaved as inhibitors of the purified GSTP1-1 enzyme, with different inhibition properties according to molecular structure. With the exception of Sulphotep showing a Ki value of 0.2 mM, all compounds reported above were characterized by high Ki values (between 2 and 16 mM) and therefore by low affinity towards GSTP1-1. These results make unlikely the use of a biosensor, based on immobilized GSTP1-1, for the detection of these molecules. On the contrary, Sulphotep can be the object of future investigations. It has to be stressed that the above listed compounds were effective on human keratinocytes, at concentrations two order of magnitude lower than that effective on purified GSTP1-1. In particular, cumene and DCB triggered a clear increase of the intracellular GSTP1-1 activity at concentrations lower than 0.1mM. These interesting results let to hypothesize the use of GSTP1-1 present in the keratinocytes as a marker for biological monitoring of workers exposed to these compounds as well as to evaluate the skin permeability of toxic compounds, not yet identified with a skin notation. PMID- 18409812 TI - [Endocrine disruptors: literature review on toxicology and application field in occupational medicine]. AB - Endocrine Disruptors (ED) and their possible impact on human health have become a topic of discussion and an area of active research in toxicology. Endocrine disrupting compound are found in plastics, detergents, surfactants, pesticides and industrial chemicals. This review will focus attention on exposure to ED in occupational context. ED are defined as an extrogenous substance or mixture that alters the function of the endocrine system and consequently causes adverse health effects in an intact organism, or its progeny. ED increases the incidence of endocrine diseases and disorders, affects the development, the immune system and influence the course of immunological disorders including autoimmune diseases. ED affects the reproductive system, they may be responsible for oligospermia, sperm characteristics, testicular steroidogenesis, testicular atrophy, uterus weight increases and proliferation of vaginal epithelium. Adverse effects are related to dose and potency of xenobiotics, ED in high doses may cause a developmental, reproductive and tumorigenic effects, however there isn't agreement that in low exposure there are effects. Symptoms from occupational exposure are reversible after exposure has end, however xenoestrogen exposure during critical stages of developmental (during pregnancy) can also result in irreversible damage. PMID- 18409813 TI - [Benzene low dose exposure: comparison of different biologic markers among exposed workers and the general population]. AB - Exposure to low doses of benzene: biological monitoring exposure in a group of occupational exposed workers and a group of general population. Even if exposure to benzene is considerably decreased in last years, the interest for this cercinogenic agent is still important. Benzene is used as additive in gasoline and it is present in cigarette smoke. Therefore benzene is considered an important toxic agent not only for occupational exposed people, but also for general population, in particular if living in high polluted areas. Aim of the present study is evaluation of trans, trans-muconic acid in urine of workers exposed to benzene and in the general population, making a distinction between subjects living in an urban centre and subjects living in rural areas. Each subject was invited to complete a questionnaire about job, diet, residence, hobbies and tobacco-smoking. Every one was submit to a physical examination, exams as ECG, audiometry, spirometry and laboratory test to investigate about blood count, liver and kidney function. It was used as biological exposure index the urinary trans, trans-muconic acid (t,t-MA). RESULTS: t,t-MA was increased in smokers compared to nonsmokers, in exposed workers (t,t-MA = 0.09 mg/L, DS 0.22) compared to general population (t,t-MA = 0.08 mg/L, DS 0.21) and in people living in urban zone compared to people living in rural areas. In particular, among occupational exposed subjects, we found concentration of urinary t,t-MA greater in the subjects living in urban zone (t,t-MA = 0.10 mg/L, DS 0.23) in comparison with the subjects living in rural areas (t,t-MA = 0.09 mg/L, DS 0.3). The same result we found in general population divided in two groups, people living in urban zone (t,t-MA = 0.09 mg/L, DS 0.21) and people living in rural areas (t,t-MA = 0.07 mg/L, DS 0.21). We have to observe that the results obtained have not a statistical relevance and the concentrations of t,t-MA in urines is always less than biologic limits. Anyway, in consideration of the carcinogenic properties of benzene and of its dose-response relation, without a treshold dose, we believe that the environmental and occupational monitoring is very important. PMID- 18409814 TI - [Matrix metalloproteinases: new effect biomarkers in the occupational exposure of toxic agents]. AB - Matrix metalloproteinases belong to a growing family of proteases controlled by specific tissue inhibitors, involved in tissutal flogosis, wound healing, cancer invasion and metastasis. We developed an in vitro model to screen for potential toxic compounds in professional exposure. Human keratinocytes (HaCat) were used as target cells while matrix metalloproteinases (MMP) were selected as responders, developing and in vitro model of allergic dermatitis. The chemical agents: potassium dichromate and nickel sulphate as positive teste, because represent the main etiological agents of allergic dermatitis. Nickel contact at very low concentrations (10(-5), 10(-6) M) induced upregulation of MMP-2 and IL-8 mRNA production; chromium contact at very low concentrations killed all cells. Actually, our in vitro research is based on analysis of cytotoxic effects of xenobiotics on human lung fibroblasts; simultaneously we verified serum increasing in vivo of MMP-9, determinated in workers serum, exposed to anesthetic gas (fluorane). In only six exposed workers we observed MMP9 increasing over than normal range. Actually, we are continuing our research on a more representative sample. PMID- 18409815 TI - [Correlation between exhaled benzene and environmental exposure in refinery workers]. AB - Benzene is still present in various working processes. Many methods are available for the biological monitoring of benzene exposure. The Authors present a study concerning a series of 23 workers of a refinery exposed to benzene. The measurement of benzene in alveolar air was preferred to quantify exposure. In this paper, adopting an original, simple and reliable device to collect the samples of alveolar air, a good correlation was found between the levels of benzene in the alveolar air and the environmental individual sampling. PMID- 18409816 TI - [Salivary analyses for biologic monitoring: possibility and perspectives]. AB - The use of saliva for biological monitoring offers several interesting advantages, in terms of cost, non-invasive sampling, no risk of infection and little likelihood of the sample being polluted. Tests for measuring chemicals in saliva are available for many endogenous analytes, and xenobiotics. Saliva tests for inorganic compounds have so far not given satisfactory results, even using highly sensitive methods such as ICP/MS or atomic absorption. However, testing saliva for pesticides holds promise: the close correlation between the plasma compartment and saliva for some of these compounds has led to proposals for assaying diazinon, trichloropyridinol, atrazine, and paraquat, for instance, in saliva as biological indicators of occupational exposure. In industrial settings too significant correlations have been found between blood, urine and saliva assays. Investigations of exposure, for example, to plastifiers (phthalates), methanol, isopropanol and m-xylene have given comparable, equally reliable data, although highly sensitive instrumentation is needed. This approach appears to offer a wide range of applications, and certainly merits further study. PMID- 18409817 TI - [Mercury toxicity: trace element interactions and detoxification activity by L arginine]. AB - Mercury (Hg) exposure makes happen disease to humans and animals spreading in all body compartments, especially in liver and kidney. In these ones, copper, zinc, manganese and iron were investigated to assess perturbation of essential metals' homeostasis due to Hg chronic intoxication. Because L-arginine, is able to induce beneficial influence on immunologic functions on mice intoxicated with Hg, we also studied the efficiency of detoxification process before and after treatment with this aminoacid. Adding L-arginine to diet of the intoxicated mice we achieved a good restoration to normal homeostatic conditions. PMID- 18409818 TI - [Biological monitoring of exposure to benzene, toluene and xylenes in urban traffic wardens by LC-MS analysis]. AB - Benzene, toluene and xylenes (BTX) are aromatic hydrocarbons widely used as industrial chemicals. Due to their formation in many combustion processes, they are widespread environmental pollutants. Thus, the general population undergoes lifelong exposure to these pollutants, and some categories of workers are exposed at particularly high levels. In order to evaluate human exposure to BTX, mercapturic acids (MA) have recently been proposed as the most specific biomarkers, but only S-phenyl mercapturic acid (S-PMA) has been validated as a biomarker for benzene. Furthermore, S-benzyl mercapturic acid (S-BMA) and S benzyl methyl mercapturic acid (S-BMMA), metabolites of toluene and o-xylene, respectively, have been less extensively investigated. In this paper, we evaluated exposure to BTX in a group of 354 traffic wardens by measuring the urinary concentration of S-PMA, S-BMA and S-BMMA using a method based on liquid chromatography-tandem mass spectrometry. This analytical method was fully validated according to U.S. Food and Drug Administration guidance. All samples showed a measurable concentration of S-BMA (mean: 12.84 microg/g (creatinine)), while S-PMA and S-BMMA resulted quantifiable only in about 30% of the analyzed urines (mean: 1.76 and 3.98 microg/g(creatinine), respectively). PMID- 18409819 TI - [Trans-muconic (t,t-MA) and S-phenilmercapturic acid: two biologic markers of benzene low dose exposure]. AB - The aim of our study is the comparison between two different biological exposure index, trans-trans-muconic acid (t,t-MA) and S-phenylmercapturic acid (S-PMA), that are minor metabolites of benzene, in the exposure to low concentrations of benzene, to estimate which shows better correlation with the environmental exposure. The study has been conducted upon a sample of 105 male workers in a petrolchemical plant in Sicily. The environmental monitoring data has shown exposure levels within acceptable limits compared with TLV, and these levels have been confirmed by biological monitoring data. The comparison between biomarkers didn't point out particular differences and this data is probably connected to low found levels of exposure. PMID- 18409820 TI - [Urinary 1-OHP excretion of urban traffic exhausts exposed workers]. AB - The vehicles exhaust in the traffic represents an important contributing factor for common air pollutants inhalation. The aim of this study was to investigate, the possible correlation between occupational exposition to polycyclic aromatic hydrocarbons (PAHs) generated from incomplete combustion, by dosage of urinary 1 hydroxypyrene (1-OHP u) as biomarker of internal dose for polycyclic aromatic hydrocarbons. In the study were included 29 workers exposed to urban traffic exhausts and of 18 non exposed workers. In boths populations was considered the tobacco smoke exposition. The results showed that exposed had levels of 1-OHP u in end-of-shift samples generally higher than those in pre-shift samples and showed an association to tobacco smoke exposition. In the control population tobacco smoke exposition appeared associated to the highest levels of 1-OHP u. The study suggests that the occupational exposure to traffic exhausts is associated with increased urinary excretion of 1-OHP. In addition, by our results seems to be a correlation between high levels of 1-OHP in the urine and increased risk of PAHs inhalation. PMID- 18409821 TI - [Genetic polymorphisms (GSTT1 e GSTM1) and urinary excretion of t,t-muconic acid among refinery workers]. AB - Many xenobiotics agents are metabolized by enzymes mechanisms through Phase I, activating substances procancerogene through oxidative reactions, and / or through mechanisms Phase II, acting on metabolic intermediate products of oxidative processes with conjugation reactions with endogenous mediators, in order to generate hydrophilic products that can be easily excreted by the body. Among the enzymes Phase II is a heterogeneous group represented by glutathione S transferase. Genetic polymorphisms encoding for these enzymes (GSTs) are responsible phenotypic expression of enzymes specifically involved in the detoxification and elimination of different genotoxic agents (IPA, toluene, benzene). Accordingly, the authors have investigated a population of subjects professionally exposed to benzene (used in active refining and storage of crude oil) in order to assess the genetic profile in relation to possible null genotype (responsible for the failure phenotypic expression of protein) of polymorphism GSTT1 and GSTM1 and correlate the impact that the genotype effect of normal metabolic pathway t, t-muconico. PMID- 18409822 TI - [Promoter effect induced by HgCl2 by studying the intercellular communication]. AB - This work aims at assessing at molecular level the effect caused by the HgCl9 intercellular communication inhibition at non-cytotoxic doses. On the basis of our previous experiences, we exposed the human keratinocytes (HUKE) at 10 nM of HgCl2 for 24 hours Next, we estimated: a) the protein expression of connexines Cx43, Cx32 and Cx26 by western blotting; b) the amount of mRNA corresponding to the three connexines by semi-quantitative RT-PCR; and c) the production of reactive oxygen species in HgCl2 treated cells using a specific probe, i.e. DCF in confocal microscopy. Our study demonstrated a higher expression of the transcripts for Cx26, Cx32, Cx43, and a higher amount of proteins Cx43, Cx32 and Cx26, compared to the negative controls. Furthermore, we studied the effect of HgCl2 on the ROS production in keratinocytes, by the analysis in confocal microscopy carried out with the DCF, fit for marking the oxygen free radicals. In HgCl2 treated keratinocytes we obtained an increase of the ROS production compared to controls; and further the mitochondrions resulted the place of ROS production. The results of this study suggest that non-cytotoxic HgCl2 concentrations, might cause an unbalancing of the redox cellular state (ROS increased level), and we can assume that the activation of a redox signalling involves the inactivation of gap junctions. PMID- 18409824 TI - [Urinary 1-naphthol in the general population of Umbria]. AB - INTRODUCTION: Naphthalene, the most volatile polycyclic aromatic hydrocarbon (PAH), was recently classified as possible human carcinogen by International Agencies for Research on Cancer Humans may be exposed to naphthalene from a wide variety of sources, including occupation, environment, personal habits. We assessed urinary excretion of 1-naphthol (1-NAF), biomarker of naphthalene exposure, in non-occupationally exposed subjects. MATERIALS AND METHODS: Urinary 1-NAF, 1-hydroxypyrene (1-OHP), biomarker of exposure to pyrene and cotinine, biomarker of smoking habits, were measured in 104 adults (53 men, 51 women). RESULTS: 1-NAF concentrations overlapped in males and females (median: men 0.35 Microg/g creat; women: 0.46 microg/g creat). Median concentration of 1-NAF was 6 fold higher in smokers compared to nonsmokers (respectively, 7.7 microg/g creatinine vs 1.3 microg/g creatinine). Between smokers, urinary cotinine was positively correlated to 1-naphthol (rho: 0.69; p < 0.01) and 1-OHP (rho: 0.53; p < 0.01). Higher 1-OHP concentrations were found in smokers (median: smokers 0.16 microg/g creatinine, not-smokers 0.05 microg/g creatinine;). CONCLUSIONS: In our study population, we found that 1-NAF excretion is much higher as compared to 1 OHP excretion. This is due to the ubiquitous presence of naphthalene in the environment. Smoking considerably increase the exposure to naftalene. PMID- 18409823 TI - [Immune disorders and chronic exposure to organic solvents. Case-control study in the operators who are in organic synthesis laboratories]. AB - The industrial society characterized by the continuing innovation in science and technology has resulted in the introduction of new business cycles chemicals which could act as "triggers" for the initiation of alterations, still latent in the clinical aspect, dependents the immune system in people who are professionally exposed. In recent years, numerous studies have shown over exposed to organic solvents some immunological abnormalities, regarding, for example: complementemy, immunoglobulinic classes, typing lymphocyte (CD3, CD4, CD8) and appearance anti-nucleo of antibodies (ANA). Our study was conducted, in health surveillance, of operators of the University of Palermo, on a sample of subjects who were exposed to organic solvents and compared with a control group not exposed, considering some parameters such as possible indicators of Immunological Trim disturbance. PMID- 18409825 TI - [Diagnostic agreement between two different analytic methods for in vitro measurement of specific IgE and comparison with clinical evaluation and in vivo test for ubiquitarian and professional allergens]. AB - This study surveys the diagnostic agreement between two different analytic systems for the measurement of serum specific IgE (sLgE), Immulite 2000 (IML) and ImmunoCap (CAP), with clinic evaluation and in vivo test. Our sample is represented by 33 adults (27 females and 6 males), with suspect of immediate allergic disease and suspect of sensitization to at least one allergen of the study panel (18 allergens: inhalant, alimentary and professional). All patients have been subjected to Skin Prick Test (SPT) for all the studied allergens, therefore sIgE measurement with IML and CAP systems has been executed (twice for each method) only for the allergens which resulted positive to the diagnostic suspect and to SPT, and for an equal number of allergens chosen by bound randomisation. In vitro methods (IML and CAP), evaluated by K Cohen statistics, give good diagnostic agreement. In the same way, the diagnostic agreement of the used analytic systems both with clinical suspect and with in vivo test is satisfying, with higher K values for IML. PMID- 18409826 TI - [Environmental exposure to inorganic lead and neurobehavioural tests among adolescents living in the Sulcis-Iglesiente, Sardinia]. AB - In several recent studies blood lead levels below 100 microg/L have been associated with reduced neurocognitive capacities and neurobehavioural performances in children, with no clear evidence of the lowest "safe" level not associated to adverse effects on the CNS. We analyzed blood lead concentration and the results of 5 neurocognitive tests, two derived from the Swedish Performance Evaluating System (SPES) and three from the Wechsler Intelligence Scale for Children-Revised (WISC-R), in 139 Sardinian adolescents living in Portoscuso, a town 2 Km far from a lead smelter, and in 72 age-matched students living in S. Antioco, a town about 15 Km far from the same smelter. The blood lead concentrations were lower than 100 microg/l in almost subjects, but, in average, significantly higher particularly among males, in the Portoscuso group compared to controls. The regression coefficients derived from the multivariate stepwise analysis, adjusted for the student's age and gender and for the educational and socio-economic level of parents, showed that reduced performances in neurocognitive test were significantly associated to increasing blood lead concentrations. According to previous our surveys in the same area and to the recent scientific literature on this topic, the present study suggests the need to further low the blood lead levels, considered by the U.S. CDC in 1991 as potentially "safe" for the children's CNS, to a threshold possibly < 50 microg/L. PMID- 18409827 TI - [Occupational exposure to hexavalent chromium during aircraft painting]. AB - Hygienists are interested in hexavalent chromium due to its genotoxic and carcinogenic effect on humans. The use of products containing hexavalent chromium is decreasing in many industrial fields because of the substitution with less toxic compounds. In the aeronautical industry, however, the chromate are added to primer paint as a corrosion inhibitor of aircrafts surfaces: so hexavalent chromium compounds are available in many primers with a composition ranging from 10% to 13%. The application of these primers by using electrostatic guns potentially exposes painting and coating workers at high concentrations of aerosols containing Cr(VI). The aim of the present study is the evaluation of professional exposure to hexavalent chromium during aircraft painting, by adopting both environmental personal sampling and biological monitoring. To valuate workers exposure levels the personal measurements results have been compared with the exposure limit values (TLV-TWA) and the urinary chromium contents with the biological exposure indices (IBE). Moreover the strategy of coupling environmental sampling with biological monitoring seems to be a useful instrument to measure the validity of the individual protection devices. PMID- 18409828 TI - [Biological monitoring in oil refinery workers]. AB - The effectiveness of t,t-muconic acid (t,t-MA) has been investigated as indicator of benzene exposure in workers of an oil refinery. 196 blue collar workers that operate in the field and 25 white collar workers who work in administration have been studied. Workers had not chronic diseases; all workers resulted fit to work. The environmental monitoring, effected with personal and fixed samplers (141 samplings), has shown that benzene air levels were lower than 170 mcg/m3. Biological monitoring was performed with measurements of blood benzene, urinary t,t-MA and urinary phenols. Each worker has been questioned regarding smoking habit, consumption of food containing sorbic acid and non-occupational exposure to benzene. Values of biological index were: t,t-MA 88 +/- 94.1 mcg/g cr, phenols 17.7 +/- 2.9 mg/g cr, blood benzene 25.4 +/- 4.3 ng/l. No significant differences were found between blue and white collars respect t,t-MA, phenols, blood benzene, non-occupational exposure to benzene, smoking habit, consumption of food containing sorbic acid. A meaningful correlation was found between the t,t-MA and the ingestion of the number of foods containing sorbic acid (r = 0.87). The results confirm that the t,t-MA seems incapable to distinguish occupationally exposed workers, at low levels of benzene exposure, from unexposed. PMID- 18409829 TI - [Assessment of the risk of arm repetitive movements among workers in the motor vehicle glass finishing industry]. AB - The aim of the study was to evaluate risk associated with biomechanical overload of the upper limbs in workers exposed to repetitive movements, employed to the finishing of glasses for motor vehicles. The risk assessment was performed using the OCRA method (OCRA index for every worker and Check-List OCRA for every workstation) and the results have been distributed for exposure levels. Altogether the results suggested the existence of risk associated with repetitive movements of the upper limbs and different risk classes (high, medium, light, very light). OCRA index and Check-List OCRA values showed together high risk in the workers with age and employment duration great (respectively 20.68% e 27.58% of workers), with especially involvement of the women, employed to jobs with high frequency. PMID- 18409830 TI - [Upper limb mechanical overcharge among health care workers]. AB - In medical circle, the disclosure of recurrent symptomatologies on the upper limbs begins to assume characteristics common to many working classes, raising not few problematic issues regarding the interactions between them and the working environment. In the AOUP of Palermo, some working categories were identified, whose activity implicates the upper limbs for instrument use, as well as repetitious movements (odontologists, ultrasonographers, endoscope technicians). The analysis covered a group of endoscopy technicians, both of the medical area and of the surgical one, the first being middle aged and with working seniority significantly higher than the latter. To these people, a RULA questionnaire was given, partially modified with the addition of some questions finalized at pointing out environmental situations that determine physical stresses on the upper limbs, correlated to the working method, to the number of made tests, to the place where they were made. From the study made, it was found that there is high risk of biomechanical overcharge on the upper limbs in the activity of endoscopy technicians. Thus the necessity of preventive corrective measures geared at avoiding on the one hand insurance claims, and on the other, the pathologies which can limit the working activity. PMID- 18409831 TI - [The participatory approach to injury prevention appeared to be an useful tool of safety education and ergonomic improvement]. AB - Participatory techniques have been adopted in vigilance of Civitavecchia's port activities, in order to control and remove injury causes. Injuries were studied by small participatory groups of port workers. As an example, seafastening operations were analyzed. The project team encouraged the workers to design ergonomic measures, or to modify work organization. Workers made suggestions and presented their points of view, that were reported to management for implementation. PMID- 18409832 TI - [Participative approach to the in hospital management of muscoloskeletal disorders]. AB - Health care workers are exposed to a wide range of musculo-skeletal hazards: manual loading (nurses, ancillary personnel, blue-collars), body posture (technicians, physicians, nurses, blue-collars), repetitive motions (clerks, blue collars, technicians). An integrated management approach to tackle musculoskeletal disorders (MSDs) was proposed in three health care organizations. An action programme, including participatory modification of tasks and jobs, and early identification and treatment of MSDs cases has been implemented, following a proper assessment of the hazards and risks. Participatory ergonomics programs proved to be effective in reducing absence rate and prevalence of workers with reduced work capacity, and in increasing workers' job satisfaction. PMID- 18409833 TI - [Low back pain in VDT operators: importance of sports activities]. AB - Video display terminal (VDT) operators are required to maintain a sitting position for extended periods of time. This transverse study assessed the prevalence of lumbar musculoskeletal disorders (low back pain, LBP) in a population of VDT operators and related results to occupational factors and other interfering variables. Data were collected using questionnaires administered to 210 public employees. LBP was defined and evaluated using the Standardized Nordic Questionnaire. The prevalence of LBP was 58.4%; it correlated significantly with the female sex, spinal disorders, sleep disturbances and stress levels. There was a significant negative correlation with exercise and sports activities and with leisure time computer use. Multiple logistic regression adjusted for age and body mass index yielded an LBP odds ratio of 0.19 (C.I.95% 0.07-0.51) for sport practicing subjects. At least one hour of exercise weekly was the cut-off for a significant reduction in LBP prevalence. The proportion of lumbar musculoskeletal symptoms was not significantly different in VDT operators and the general population. None of the occupational variables considered correlated with development of LBP. The occupational physician should recommend VDT operators to devote some leisure time to exercise. PMID- 18409834 TI - [The Italian version of Nordic Musculoskeletal Standardized Questionnaire]. AB - We translated into Italian the Nordic musculoskelethal questionnaire, as completed by Canadian IRSST with Authors' agreement in 2001, according to OMS recommendations. This translation involved the following items: aches and troubles of neck, dorsal region, low back, shoulders, elbows, hands and wrists, hips and thighs, ankles and feet in the last 12 months. The questionnaire was then submitted to reliability and stability tests. The Italian version of the questionnaire, already used in different languages, proved to be suitable and reliable also for self administration. PMID- 18409835 TI - [Evaluating patients manual handling hazards in nursing personnel of a hospital]. AB - Low back pain (LBP) and shoulder and arm disorders are common among workers exposed to risk due to manual weight lifting, and this health care problem is also very costly. We also know that nursing personnel exposed to manual patient handling activity report high number of work-related musculoskeletal disorders. The objective of our study was to evaluate the relationships between work and musculoskeletal disorders in personnel exposed to manual patient handling activity. 160 health care workers of an hospice exposed to patient handling were compared to a control group of 172 people not exposed to the handling risk. In our study we considered only subject with pathology already diagnosed, withdrawing people with disturbs but without clinical trials. The statistical evaluation using the chi2 test has not shown any meaning in the comparison between the two groups, we have calculated the Odds Ratio risk for discal hernia and protrusion finding a risk between low and modest (1.52). We think that the least prevalence of musculoskeletal disorders to the back in the group exposed to manual patient handling activity, could be explained partly with the "healthy worker" effect and partly with the workplace improvement. That is demonstrated also with the M.A.P.O. index scoring between 0 and 1.5 (negligible risk). PMID- 18409836 TI - [Whole-body vibration risk among operators in railway engines shunting ]. AB - Purpose of the present note is to assess the risk from Whole-body vibration (WBV) in operators employed in the shunting of engines within the railway stations. The study has been conducted in the cockpits of the shunting engines used within the railway station of Villa S. Giovanni (RC). The measures have been taken through accelerometer IHVM 100 Larson-Davis, placed on the seat of each locomotives for a recording time of around 15 minutes. A standard measure has been effected besides, positioning the sensor on the floor of the same locomotives. The measurements indicate that the risk to these workers is negligible because in any case the value is exceeded action daily 0.5 m/s2, having recorded values range from 0.1 to 0.2 m / s2. In conclusion it holds him necessary, to the preventive goals, in respect to how much anticipated from the D.L.gs 187/05 the necessary technical, organizational and formative measures to the containment of the risk. PMID- 18409837 TI - [The risk of manual handling loads in the hotel sector]. AB - The aim of our study is to evaluate the manual handling risk and the incidence of muscle-skeletal pathologies in the hotel compartment. Our study is conducted on 264 workers of the hotel compartment. The sample is divided on the base of the working turn in the following groups: porter (both to the plans and in the kitchen); waiters to the plans; services (gardeners and workers). The duties have been valued according to the method NIOSH. The presence of muscle-skeletal pathologies has been verified on the base to the accused symptomology, and on the presence of clinical objectivity and to the reports of checks. The data has been compared to a control group. The application of the NIOSH method has showed for each working profile an elevated synthetic index, > 3, and for porter the index is 5. The clinical data has shown an elevated incidence of pathologies of the spine, especially lumbar spine, with a high prevalence in the group of male porters. In conclusion we believe that the manual handling represents a particularly remarkable risk for the workers in the hotel compartment. PMID- 18409838 TI - [Exposure to whole body vibrations in workers moving heavy items by mechanical vehicles in the warehouse of a large retail outlet]. AB - Efficient warehouse management and item transportation is of fundamental importance in the commercial outlet in exam. Whole body vibrations have been measured in various types of machines, some of which not widely studied yet, like the electrical pallet truck. In some tasks (fork lifts drivers) vibrations propagate through the driving seat whereas in some other tasks (electrical pallet trucks, stackers), operated in a standing posture, vibrations propagate through the lower limbs. Results have been provided for a homogeneous job tasks. In particular conditions, the action level of the Italian national (and European) regulations on occupational exposure to WBV may be exceeded. The authors propose a simple system of probabilistic classification of the risk of exposure to whole body vibrations, based on the respective areas of the distribution which lay within the three risk classes. PMID- 18409839 TI - [Acute lumbago prevalence of health workers exposed to a moderate level of exposure index MAPO]. AB - We conduced a cross sectional study in the period Gen - Dec 2006 to examine the relationship between acute lumbago in health workers and exposition to a moderate level of exposure index Movement and Assistance of Hospital Patients (MAPO). The study ruled 240 health workers (M: 180; F: 60), the mean age was 44,9 years (range 24 - 64); was evaluated the occurrence of acute lumbago in the last 12 months. The objective of this study was to describe the trends over a specific time of the association between the moderate MAPO index and acute lumbago in this sample of health care workers. The results indicate that healthcare workers exposed to moderate MAPO index appear to incur a greater risk of acute lumbago than general population, but lower than that evidenced by Italian and Sweden authors between nursing staffs. Medical surveillance of exposed workers is confirmed as necessary method of secondary prevention and also it is useful in the diagnosis of worker's susceptibilities. PMID- 18409840 TI - [Biomechanical risk assessment of manual material handling in vegetables and fruit departments of supermarkets]. AB - There is little in the literature about the risks of manual handling of material in supermarkets and what there is refers solely to storehouse work. This contrasts with the substantial number of studies of the risk of repeated arm movements among supermarket cash-desk staff. The scarcity of information is partly due to the difficulties of applying widely employed, standardized evaluation methods in this sector. One of the conditions limiting the application of the NIOSH protocol in this retail sector is that lifting tasks are so often closely tied to transport. The biomechanical analysis method we used brought to light considerable risks in many of the steps investigated: unpacking the pallet, unloading the crates from the pallet to the ground, lifting them from the floor onto display stands, and filling the boxes on the stands with goods before the shop opens. Images acquired on site were analyzed in the laboratory. We selected the most indicative images, which were then studied as regards posture and biomechanics using Apalys 3.0 software (ILMCAD GmbH, Ilmenau, Germany). Biomechemical analysis was done on the following movements: unloading crates from the pallet, positioning them on fruit and vegetable department display stands, and filling the boxes on the stands. We obtained a prediction of 2720 to 5472 N for the load at the lumbosacral junction (L5-S1). Simulation of the NIOSH index gave a value of 2.69 in the only case where the Waters protocol could be applied. PMID- 18409841 TI - [Usefulness of surface electromyography of hand muscles in the assessment of myoelectric parameters changes due to repetitive manual tasks]. AB - The aim of this project was to investigate the possible role of sEMG in the diagnosis of Carpal tunnel syndrome (CTS). The study group consisted of 37 subjects, of whom 14 (control group) were not employed in manual tasks and 23 (exposed) were engaged in repetitive and forceful manual tasks. Of the 23 exposed workers, 10 reported CTS symptoms, whereas all the subjects of the control group resulted asymptomatic. The surface electromyography signal was recorded from the abductor pollicis brevis muscle, using different levels of isometric contraction: 20% and 50% of the maximum voluntary contraction (MVC), respectively. The initial values and rate of change of the average rectified value (ARV), mean power spectral frequency (MNF) and conduction velocity (CV) were calculated. Moreover the study protocol included clinical evaluation and electrodiagnostic study of the median nerve. Data from the exposed and control group were compared. Statistically significant differences between the two groups were found for ARV initial value and for CV and MNF rate of change at 50% MVC. These parameters resulted lower in the exposed group, with the lowest values among symptomatic subjects. Possible explanations may be the loss of motor units, particularly affecting the fast and fatigable type II muscle fibers, involved in the myoelectric manifestation of fatigue. In conclusion, this technique was able to show different myoelectric patterns and manifestations of fatigue between subjects exposed and nonexposed to manual intensive work, suffering or not suffering from CTS. PMID- 18409842 TI - [Function analysis of employees at cash desks in the retailing sector by means of the study of the movements of the upper limbs]. AB - Supermarket checkouts are different in term of dimensions and arrangement of the essential devices for cashier's work. It means that a different layout can involve different tasks, so it is necessary to evaluate every single system in order to define the entity of required commitment for the worker. A supermarket checkout was considered in order to evaluate its workstation and to analyze the different tasks of two cashiers with similar anthropometric measurements. In particular one of them is affected by a widespread osteoarthritis, verified by instrumental tests and clinical examination. Several geometrical measures of the workstation were carried out while the cashiers were observed and filmed by videotapes during their activities. The study has been carried out by drawing the "preferred" and "maximal" working areas, as UNI EN ISO 14738:2004 suggests. Some tasks seem to be particularly critical especially for the cashier affected by osteoarthritis, who adopts compensatory postural mechanisms in order to use some devices which are not easy to reach inside the workstation, and which can induce pain to her. Finally the authors make some considerations about the opportunity of making a new layout of the workstation in order to find a more suitable arrangement of the devices. PMID- 18409843 TI - [LBP and disability in nursing personnel performing manual handing of patients in a large Italian hospital]. AB - Low back pain (LBP) has been recognized as a major cause of sickness and disability among health care workers (HCW) who perform handling of patients. A survey was carrying out to investigate risk factors for LBP and related absenteeism and disability in a group of HCW. 333 HCW were examined in 2003 and 2005 and data were collected on personal, occupational and clinical history and on risk factors for LBP and related absenteeism. Psychosocial factors and LBP related disability were evaluated by means of Karasek's Job Content questionnaire and Oswestry Low Back disability Index. Most of our study population were female (80%) and the prevalence of acute and chronic LBP was 37%. Subjects with chronic LBP referred a significantly higher level of disability as compared to acute LBP subjects. Moreover, a significantly relationship was found (p = 0.04) between high level of disability (group III e IV of ODI index) and high perceived job demand. The introduction of questionnaires to collect disability and psychosocial factors is highly recommended in periodic health surveillance program of HCW performing manual handling of patients. PMID- 18409844 TI - [A survey on musculoskeletal disorders in physiotherapists]. AB - Only few studies focused on musculoskeletal disorders (MSD) among physiotherapists (PT). The study population consisted of 50 PT and 50 clerical workers belonging to the same three rehabilitation hospitals in Northern Italy. The participants filled in a specifically designed questionnaire focused on risk factors, symptoms and diseases related to MSD. Data were self reported. Preliminary data show a significantly higher prevalence of low back symptoms (70%) and upper limb symptoms (36%) in PT than controls (16% and 4% respectively). PT had a higher prevalence of lumbar disk degeneration (20%), shoulder disorders (14%), wrist and hand tendinopathy (10%) than clerical workers (4% and 2% respectively). Both PT and clerical workers attributed their cervical symptoms to work (85% and 95% respectively). Only PT believed that low back and upper limb symptoms were work related (83% and 94% respectively). Our survey supported the findings that PT have e high prevalence of MSD. Specific preventive intervention should be applied to reduce risk of work related MSD. PMID- 18409845 TI - [Theoretical-practical training for personnel involved in patient assistance for limiting the risk of manual handling of the patients in a hospital company]. AB - Worker education and training must represent a deep cultural change that can be achieved, fulfilled and strengthened through constant guiding steps and periodical checks regarding practical application of acquired knowledge. In our hospital, a multi-subject team has been identified to face such problem of risk reduction from inpatient mobilization in a shared and synergic manner. Therefore a theory and practical training course is being proposed, with a theoretic part of one hour and a half to be held in a classroom to small groups (not more than 20 people) and a practical part (addressed to 8 people groups) composed of 4 sections of 1 hour and a half each: once listening in class and then being subdivided in the relative belonging wards as well as after an explanation, everyone will have here the chance to perform the correct manual mobilization handlings also in association with the use of minor aid devices. At the end of each section, a referent-educator is identified for each Operative Unit: such a figure is extremely important as a reference for his/her collegues, for new employees and to support and/or substitute the outside inspector in checking operations during the year following the end of the course itself. This latest phase is thus considered absolutely necessary to realize the actual change in the staff habits. PMID- 18409846 TI - [Work related low-back pain: prevention and strategies of rehabilitation]. AB - Back disorders are a major health problem in many occupations. Physical loads at work, such as manual materials handling frequent bending and twisting, lifting, and forceful movements, have been identified as possible risk factors for low back pain (LBP). Aim of the present study was to evaluate the rehabilitative treatment in the chronic lumbago. PMID- 18409847 TI - [Prevention of low back pain diseases in a center for the disabled]. AB - Our unit of occupational medicine has been asked to solve the problem of an incrising percentage of workers affected by low-back pain in a clinic for patients with handicaps. The consequence of this affections caused frequently absences at work. We organized a course for training about correct methods for moving patient not self-sufficient. The 84 workers, all female (aged 30-45), had to sign on a card all the problems they found at work and their possible solutions according to them. Then we put the cards in a box and we red each one, signing on a blackboard the problems, using a red pen and the solutions proposed, in blue. Bed organization, turnation of patient, and the difficulties in the use of lifter were the principal problems, solved trought the training course made. Then we obtained a reduction in absence at work for low-back pain of about 40% after six months since the beginning of a course of posture exercises made in the same clinic one day a week after work. PMID- 18409848 TI - [Upper limb work-related disorders: description of an Italian hospital based case study]. AB - Using the Hospital computerized database, individual variables including referral source, gender; age, life style, work sector, clinical and work history, final clinical diagnosis and work-relatedness evaluation were retrieved and analysed. The results of this survey showed a significant increase of number of cases, especially from 1999 to 2006. Shoulder disorders showed a significant increase in the latest years of the observation period, while the other upper limb districts remained constant. The work sectors mostly affected were textile, construction, metalmechanical and food industry. The findings of this study indicates an increasing demand for clinical assessment of work-related musculoskeletal disorders of the upper limb. Therefore, it is important that a standardized diagnostic procedure is applied together with an accurate evaluation of biomechanical overload. PMID- 18409849 TI - [Hand-arm vibration syndrome and upper limbs diseases in the forest workers of Italia meridionale]. AB - Vibration exposure of the hand-arm system is associated with an increased risk of upper-limb vascular, neurological and musculoskeletal lesions, or hand-arm vibration syndrome (HAVS). The prevalence of occupational HAVS and upper-limb disorders was studied among 278 Forestry Service workers in Sicily and Calabria. Subjects who used chain-saws (18 weeks/year) had a greater prevalence of peripheral sensory-neural disturbances (28%), upper-limb musculoskeletal disorders (33%) and carpal tunnel syndrome (19%) compared with 260 manual workers from the same Corps not exposed to hand-transmitted vibration. Raynaud's phenomenon was comparable in exposed and control subjects (5.3% vs. 4.7%.) Upper limb neuropathies were significantly associated with energy-equivalent frequency weighted acceleration; exposure duration; and cumulative vibration dose (m2/s4h). The variable "years of work with vibrating tools" was strongly associated with peripheral neuropathies; carpal tunnel syndrome; and upper-limb musculotendinous syndromes. Data suggest that in Sicily and Calabria, where the climate is milder than in other areas of Italy, forestry work with hand-held vibrating tools does not entail a greater prevalence of peripheral vascular disorders (Raynaud's phenomenon), while the prevalence of occupational upper-limb neurological and musculoskeletal disorders, in which combined ergonomic and mechanical risk factors have a large pathogenic role, is significantly increased. PMID- 18409850 TI - [Cervical, thoracic and upper limb load assessment in dental professionals]. AB - In order to analyze the effect of working posture on the level of neck, back and shoulder muscular activity, a clinico-anamnestic, electromyographic and postural study of 34 dentistry was undertaken. Using surface electrodes, the level of muscular activity was recorded. The results showed that neck flexed working posture is prevalent: using stereomicroscope gave lower levels than with or without enlarging systems, such as glasses. PMID- 18409851 TI - [Risk assessment in upper limb overload]. AB - One of the most important factors of the work-related musculoskeletal disorders of the upper extremities (WMSDs) is the biomechanical overload. The purpose of this study is to evaluate the possibility to predict the upper limb repetitive stress, according to risk assessment procedures. In order to this aim, we gathered clinical-anamnestic data and risk assessment considerations of a cohort of workers in a car industry. PMID- 18409852 TI - [Low back pain, movement impairement syndromes classification]. AB - According to literature up to 85 percent of low back pain (LBP) cannot match a precise pathoanatomical diagnosis, particularly when the association between symptoms and imaging is weak. Effective diagnostic strategy is mainly needed by occupational and rehabilitation medicine. The present study is based on the hypothesis that cumulative effect of repetitive movements and sustained postures, particularly when the movement deviates from the optimal kinesiologic standard, can lead to tissue damage. Therefore this cross-sectional study answers to lack of standardization for LBP diagnosis by means of the classification proposed from Sahrmann within the Movement Impairment Syndromes (MSI) concept. The classification defines 5 categories of LBP: flexion, extension, rotation, extension-rotation and flexion-rotation. The present study results on 84 health workers with LBP match previous published data on the prevalence of MSI categories: 54.8% extension-rotation, 13% flexion-rotation, 11.9% rotation, 3.5% and 16.6% unclassified. MSI could also contribute to better understand LBP severity and the following restricted duty evaluation. Further studies to increase the sample size and to set up randomized clinical trials are needed. PMID- 18409853 TI - [Professional exposure to hardwood dusts in a group of Sicilian joiners]. AB - Wood is a system of vegetable tissues chemically formed by biopolymers (90-99%) and phenolic substances, alkaloids, glucosides and saponins (1-10%). There are two botanical groups of wood: hardwood and softwood. Aim of present study was the research of possible work related diseases with professional exposure to hardwood about 25 joiners of Palermo in 2006; in fact wood dust inhalation is very dangerous for workers health because it is cancerogenous for nasal and paranasal sinus (1 IARC). We studied beech and fraxinus dusts (hardwood) and mahogany and teak dusts (exotic hardwood) in the environmental air of joiners during wood cutting and finishing touch. We also analysed exposed workers registry to know joiners duties, exposure values to wood dust, inhalatory exposure, wood type, chemical components, individual protection safety devices, environmental protection means. Then we examined workers medical case and risk histories. At the end we noticed that particle material in environmental air was regular according law reference values (D.Lgs. 66/2000: TLV-TWA 5 mg/m3); but although environmental dust values were normal, we found nasal haemorrhages in 3 workers and vasomotory headache in other 3; we also diagnosed one paranasal sinus polypus and then irritative contact dermatitis in 3 workers. So there is an answer: "can current TLV-TWA be protective and sure? And can it guarantee health of hardwood exposed workers?". PMID- 18409854 TI - [Wood dust exposure in handicraft companies in Lazio: preliminary findings]. AB - The aim of this study is to evaluate the wood dust exposures characterizing the working more to risk. Two selectors for inhalable fraction were been used: IOM sampler (Institute Occupational Medicine) and conical sampler. The sampling time is choosen by environmental airborne dust and it has varied around three and four hours. The surveys involved some companies of the Lazio that carried out the second working of wood that predict the transformation in ultimate products. The woodworking processes investigated have been grouped in three different classes: wood dissection; planning and moulding; sanding. The results have shown that the medium concentration of wood dusts, obtained in three processes investigated ranges from 2 mg/m3 to 4 mg/m3. The higher amounts is of 16 mg/m3 and it have been obtained during the sanding. Moreover the dust collected by IOM sampler was always higher than by conical sampler probably it is due to large particulates that were projected into IOM causing an overestimate of the amount of wood dust particulate that was inhalable. This study need to of more personal sampling for being able to carry out an exhaustive statistical analysis. PMID- 18409855 TI - [Occupational diseases in agricultural workers 2002-2006]. AB - The authors have examined the data of the occupational diseases in agriculture in the period 2002-2006; such data evidence as in these years there has been an increase of the number of the denunciations of occupational diseases. The increase is referred to new emergent diseases" as occupational the syndrome of the carpal tunnel and the tendinitises. The acknowledgment of new occupational diseases must be accompanied also by an effective promotion of the security of workers. PMID- 18409856 TI - [Professional diseases in the handicraft's workers 2002-2000]. AB - The authors have examined the data supplied from the Actuarial Advising Statistics of the INAIL about occupational diseases in the handicraft's workers denounced and recognized in the period 2002 -2006. The data have been analysed by sex, age, nature and site of lesion. PMID- 18409857 TI - [Alcohol related risk survey about health workers]. AB - The link between drinking and consequent injuries during job activities has been stresses by the most important international organizations and by ISTAT. Luckily, the low 125/01 has opened to "occupational doctor" the possibility of analysing the alcohol related risks among particular working classes, i.e. hospital workers. Thus, it has been conducted a survey in our Hospital in order to understand the entity of the alcohol related risk. The 32.59 of the sample was classified as teetotallers, 45.81% was in the range of low risk probability, while the 24.58% was in the high risk one. It has also been noticed a significant link between the highest level of risk and male employees, but nothing in relation with age or the type of job. In conclusion, this study has represented the first step for our next training programs to reduce the alcohol related risks and give a support to those workers who have already drinking problems. PMID- 18409858 TI - [Epidemiology of work-related diseases in ULSS 12 Venice (Italy)]. AB - In Veneto, like in Italy, in the last years the course of the professional diseases shown a trend in reduction. This trend has had to the difficulty to recognize the professional aetiology of multifactorial diseases. In the Venice the analysis of the course of the professional diseases in last the 4-5 years has demonstrated an increase of the communications of diseases from the doctors who operate in hospital to the SPISAL for the active search for pathologies asbestos and CVM correlated; moreover it has been a reduction of the hearing loss from noise from 2000 and it has been increment of cancer of lung and mesothelioma from 2001. Emergent diseases, like the allergy, the back diseases and those tied to the organizational constriction, are sottostimate. They have been a collaboration with the doctors of hospital, the doctors of factories, the INAIL and the court. PMID- 18409859 TI - [Work related diseases: assistance to delivery among obstetricians]. AB - In the course of Sanitary Surveillance, a high incidence of pathologies of the superior limbs was encountered in a homogeneous group of midwives operating in the AOUP of Palermo. These findings induced the authors' starting a retrospective study aimed at understanding the possibility of correlation between work effected and damage to people, and geared at being able to frame the findings in the ambit of the related to occupational pathologies. It is a fact that the positions taken by midwives in the course of gynecological vaginal delivery, or those necessary to the maneuvers during childbirth, such as the lateral or the simple and/or exaggerated MacRoberts maneuver, require substantial overcharging of the spinal column and a significant engagement of the muscles of the upper limbs. The analysis of the data related to the working load effected in the various years (number of parts/years), highlighted how the group of midwives exposed to a higher risk were those with a longer working life, this in consideration of the great emphasis placed on the upper limbs. It cannot therefore be ruled out that the pathologies encountered in the group of midwives can be framed in the area of occupational related pathologies. PMID- 18409860 TI - [Prevalence and risk factors of urinary incontinence in female workers of hotels]. AB - The International Continence Society defines urinary incontinence (UI) as "a condition in which involuntary loss of urine is a social or hygienic problem and is objectively demonstrable". There are three different jorms of UI. stress urinary incontinence, urge urinary incontinence and mixed incontinence. The aim of this study was to investigate the prevalence of UI in a group of female workers in the hotel sector. The International Consultation on Incontinence Questionnaire Urinary Incontinence short form (ICIQ-UI Short Form) was administered to all female workers and data were collected about age, body mass index, number of vaginal and Caesarean delivery. Results showed a prevalence of UI widely bigger in the plans waitress than in video display terminal workers and suggest the hypothesis that manual handling of loads representing a possible occupational risk for UI. PMID- 18409861 TI - [Evaluation of noise risk in roadmen]. AB - The activity of roadman is not much knowed. The evalutation of the risk evidences the noise and vibrations as the principal polluter. The aim of our study is to go into the knowledge of the working risk of that job. The study was leaded on a sample of 287 roadmen. It was carried out an evalutation of noise exposure of the workers and a biologic evaluation that included: Medical check. Othorhin.laryngoiatric examination. Audiometric examination. Impedenzometric examination. The audiometric exam was conduced in a silent box and with a acoustic rest at least 16 h. The audiometric test were classified in conformity with Merluzzi-Pira-Bosio method. The phonometric evaluations point out, in roadmans examinated, hight exposure to noise with mean Lep d > 90 db. Biological evaluation, through medical check, showed data similar with that of a general group homogeneous for age and sex. Audiometric evaluation showed a noise acoustic trama in 60% of sample. Inpedenzometric exam confirmed the absence of transmissive damage in almost whole sample. In conclusion we can adfirm that noise is a strong factor for roadmen, with evident damage to exposed workers. PMID- 18409862 TI - [Prevalence of occupational allergic symptoms among bakers of Verona]. AB - The aim of the study is to evaluate the prevalence of respiratory and skin symptoms among bakers in the district of Verona by means of two standardized questionnaires, a clinical one administered to the bakeries employees, either directly exposed to flour dust or not exposed, and one concerning the characteristics of the bakery environment. The study involved 613 bakery workers, 401 bakers and 212 sales personnel employed in 152 bakeries. Bakers exposed to flour dust reported nasal symptoms in 29.2% of cases, ocular symptoms in 13.2%, cough, shortness of breath, chest tightness, wheezing respectively in 8.7%, 7%, 4.5% 6.2% of cases, itch and skin burning with paleness respectively in 9.7% and 8.5%. A significantly lower prevalence of these symptoms, except skin paleness, was reported among the sales personnel. In bakers, a significant correlation was found between ocular-nasal and respiratory symptoms and family or self-reported history of atopy (p = 0.001). Reported flour dust air pollution at the workplace correlated with occupational ocular-nasal and respiratory symptoms (p = 0.001). Flour dust on skin and clothes correlated with crusts and excoriations (p = 0.01). Our study confirms the high prevalence of allergic symptoms among bakers and the need of clinical assessment. PMID- 18409863 TI - [Diagnosis and insurance compensation of occupational diseases in construction industry]. AB - The aim of this study is to evaluate the outcome of 302 occupational diseases in building workers detected by UOOML Ospedali Riuniti of Bergamo and notified to INAIL from 2000 to 2005. The 41.3% of cases were accepted as work-related. Among remaining cases (58.7%), INAIL rejected 40.9% for lack or absence of documentation. 59.1% for no adhesion to legal medicine criteria. By analysis of occupational diseases detected in the last 5 years, we found an increase of muscle-skeletal disorders, for which, now, diagnostic procedure and insurance evaluation are difficult. This work shows a wide gap between reported occupational diseases of buildings workers and compensation given by INAIL. These results underlines the need of comparison among involved institutions in order to standardize statistical and diagnostic instruments. PMID- 18409864 TI - [Incidence of dysphonia in teaching staff of schools]. AB - Some categories of workers (actors, teachers, singers) use a particular working tool: their voice. In order to assess the spread of dysphonia and its possible causes across a specific class of workers--the teachers of some schools in the city of Catania--we have set up a ad hoc questionnaire. A special database has been subsequently arranged, aimed at the statistical analysis of the responses obtained. The analysis of the responses shows that the phenomenon is more present in female than male teachers, 48% versus 30%. The dysphonia occurs mainly at the end of the working day, lasts throughout the school year and improves in summer, demonstrating the importance of teaching activity in its pathogenesis. One aspect especially stands out: dysphonia occurs exclusively in those individuals suffering from diseases with possible alterations of the voice, such as allergopathies, tireopathies and gastropathies. Therefore, work activity alone does not seem able to cause a chronic dysphonia, whereas the presence of diseases causing disability of vocal cords appears to play an important role. PMID- 18409865 TI - [Chronic-degenerative diseases in health care activities: data from health surveillance of Federico II University General Hospital in Naples]. AB - Recent average life increase, as well as new habits and lifestyles assumption, has determined a change in population health profiles, as a result of progressive increase in chronic-degenerative diseases prevalence. Among these latter; musculoskeletal and cardiovascular diseases are the most frequent ones. Health care workers also suffered from this change in health profiles. This study, performed thanks to the extension of Health Surveillance to all health care workers, aimed at evaluating chronic-degenerative diseases risk among three main welfare activities (medical, nursing and auxiliary). In conclusion, data showed that nurses, mainly the shift workers, are exposed to a higher risk of chronic degenerative diseases. All other health care workers have not significant work risk factors, because of a greater burden of aging and lifestyles effects. PMID- 18409866 TI - [Exposure to pesticides and non-Hodgkin lymphoma. A meta-analysis of observational studies]. PMID- 18409867 TI - [Have the prevalence and incidence of occupational asthma and rhinitis due to laboratory animals declined in the last 25 years?]. AB - Data for time trends in the prevalence of occupational asthma (OA) and rhinitis (OR) are not known. To investigate trends in the prevalence and incidence of OA and OR over time, we reviewed the available literature. We analysed 15 cross sectional and 4 longitudinal studies published from 1980 to 2006. In workers exposed to laboratory animals (LA) the prevalence of OA ranged from 2.2% to 11.7%; the prevalence of OR ranged from 6.7% to 41.7%. When we analysed prevalence by study date using a logistic model, the estimated prevalence of OA declined from 8.7% in 1976 to 4.2% in 2001 (p < 0.003), which is -1.8% every ten years. The prevalence of OR rose slightly from 18.5% in 1976 to 19.7% in 2001 (NS). In four longitudinal studies the incidence of OA ranged from 0.4 to 3.5/100 person years, while the incidence of OR ranged from 2.0 to 10.1/100 person years. No clear trend emerged over time. This review shows a trend toward a progressive decline in the prevalence of occupational asthma due to laboratory animals, which may be due to the reduction of exposure since the early 1980s. A further reduction of exposure is needed to prevent the onset of occupational rhinitis. PMID- 18409868 TI - [Safety of professional drivers: literature review about prevention measures linked to sleeping]. AB - This study assess the need of development of a common protocol to detect occupational driver with high risk of driving accidents. Several studies have indicated that excessive sleepiness and daytime alertness cause an increased risk of driving accidents in occupational drivers. Excessive daytime sleepiness affects 4 to 12% of general population. The 37% of drivers reported that they have nodded off or fallen asleep at least once in their driving career. A substantial percentage of drivers suffer from sleep-disordered breathing, narcolepsy, sleep deprivation, poor sleep hygiene, changes in wakefulness-sleep circadian rhythm. Some important factors in determining driver sleepiness are duration of the drive, driving at night, more experience as driver, arduous schedules. There is a lack of awareness among the public and among physicians and authorities in general of the problem posed by sleepy drivers. There isn't a national surveillance system that identify sleep loss or sleepiness in relation to work. The first target of a screening program protocol should be to detect the presence of obstructive sleep apnea. Overnight polysomnography in a sleep laboratory is normal practice for diagnosing obstructive sleep apnea but it is expensive. The ambulatory approach can be used to expedite management of screening in occupational driver. PMID- 18409870 TI - [Analysis of accidents at work in building yards: correlation between risk and law defaults in yards of Palermo]. AB - In the last years the building firm underwent real boom for investments and workers but not for safety measures. In the building sector there is a great accidents risks for falls from the top. The aim of present study is to analyze trend of accidents at work in building yards according to INAIL DATA and we will identify causes and risks then we will correlate these risks with frequent law defaults that we noticed in the province of Palermo. At the end we will value possible preventive measures to avoid this phenomenon. DATA INAIL of period 2001 2005 about building sector show us a reduction of denounced accidents at work but this phenomenon is changeable. Palermo is the first sicilian province for accidents at work although the number of denounced accidents (from 2001 to 2005) is decreasing while this number in Catania is increasing, CPT of Palermo gave us data (from 2004 to 2006) about inspections in building yards. Our study interests 609 yards and 1827 work stages. We noticed the most number of rests in civil yards. PMID- 18409869 TI - [Incidence of occupational accident in Sicilian University Hospital]. AB - The evolution of the disease accident has been analysed in a University Hospital in Sicily (approx. 2500 employees/year) for the time period between 2002 and 2006. The industrial injuries index has shown a swing in course (from a value of 3.80% in 2002 to 5.17% in 2003 to 4.49% in 2004), affecting the categories of nurses and health social workers most. The phenomenon shows a constant course within the same period for medical staff, technical assistants and administrative staff. Biological risk industrial injuries have the highest frequency compared to all others (62.5% in 2002 down to 42.3% in 2006). The most interesting datum is the relative scarce entity of the phenomenon when compared to other similar hospital situations and the progressive reduction of biological risk incidents. PMID- 18409871 TI - [Accidents at work in agriculture workers in the period 2002-2006]. AB - The authors consider the accidents happened in agricultural environment in the period from 2002 to 2006. The data of the Inail evidence a decrease of the denounced events, but with a reduction of the occupied workers. Such data make demonstrate that not it there has not been an improvement. For this reason it's necessary put in action informative and formative campaigns about the culture of the prevention of agricultural accidents. PMID- 18409872 TI - [Accidents at work in handicraft workers in the period 2002-2006]. AB - The authors consider the phenomenon of the accidents at work in the handicraft workers in the last period; the data examined demonstrate that in this work environment there has not been an evident decrement of the cases in comparison to the past. It's however always necessary to promote and improve the preventional measures in these particular job places. PMID- 18409873 TI - [Nocturnal accidents in quinquennium 2002-2006]. AB - The incidence of nocturnal work accidents were examined using the data bank of Italian Workers Compensation Authority. Analysis carried out leads us to conclude that the nocturnal accidents are not more frequent than the total events. Finally nocturnal events are more dangerous and the frequency of fatal accidents in the night is higher if it is compared to that of diurnal period. PMID- 18409874 TI - [Accidents at "work in extracomunitarian workers" 2002-2006]. AB - The authors illustrate the phenomenon of accidents at work of extracommunitarian workers who work in our country. The aim is to activate competent to institution and demonstrate the so-called "black job" and to establish adequate measures apt to prevent the accidents at work for these workers. PMID- 18409875 TI - [Reactive anxiety crisis: a unique case of work injury]. AB - The present study aims to describe a unique case in view of the disease diagnosed, the conditions of onset and the management by INAIL (Italian National Institute of Insurance for Injuries at Work and Occupational Diseases). A worker, after a verbal, animated dispute with some collegues, had an acute psychiatric agitation attack and went to the nearest emergency room, where he was investigated. No neuropsychiatric alteration was diagnosed, but based on anamnestic data, the physicians diagnosed an anxiety crisis reactive to work environment. A medical certificate for injury at work was produced and sent to the Insurance Board (INAIL). The worker was off work for 105 days diagnosed with a persistent anxious depressive syndrome, secondary to the traumatic event. INAIL reassessed the case later and confirmed only the first 30 days as due to work accident, while the following period was judged as related to affectivity disturbance due to common disease, not to work environment. Our case opens new perspective for the occupational physician in the assessment of ASD as work injury and of PTSD as professional disease, suggesting to give more attention to psychiatric health of workers. PMID- 18409876 TI - [Needlestick and sharps injuries among nursing students: an emerging occupational risk]. AB - The biohazard represents a major occupational risk among workers in the health sector, this risk is not only exclusive for healthcare workers but involve also nursing students. The study reports data of a survey on injuries from accidental puncture in a group of 223 students of the third year of Nursing of Bari University. The 18% of students say they have suffered over the past 12 months an accidental puncture with sharp instruments. The cutting device most frequently involved is the needle from the syringe and insulin. The most at risk are the recovered and disposal of the needle. The biohazard in training is further compounded by factors such as lack of experience and skill manuals consolidated combined with a non perception of the risk. In the obligation of protection, training and information to students of Nursing, the University must implement programs aimed at both knowledge of the risks to which they are exposed, as well as security procedures to contain an emerging risk, which one of injury from sharp instruments, which are exposed young students not yet in employment. PMID- 18409877 TI - [Accidents at work in construction workers: a peculiar utilization of the information flow INAIL-ISPESL-Regions]. AB - Indicators generally used to describe the work accident risk, i.e. frequency and injuries severity, are not suitable for comparison among single companies in order to point out those with major risk. We propose a method to draw out construction companies with higher risk of occupational injuries in the period 2003-2005 from INAIL-ISPESL-Regioni database. These companies will be object of specific interventions for the prevention, training and supervision from Prevention and Safety in Working Environments Services, from Provincial Labor Directions and Provincial Territorial Construction Committees. PMID- 18409878 TI - [Retrospective analysis of the accidents causing work inability in a teaching hospital during the time 2002-2006]. AB - This study is a retrospective inquiry, based on the data of University Hospital "L. Sacco" in the period of five years 2002-2006. They were analyzed the relevant data to beyond 350 events cause of working inability. PMID- 18409879 TI - [Occupational injuries in nursing school students]. AB - Occupational injuries represent an important risk factor in the nurses. In this paper we have studied the characteristics of the phenomenon in a group of nursing school students of one University of the Lazio. The results show an elevated frequency of the phenomenon, characterized exclusively from biological accidents and the necessity to plan preventive measures, insisting, mainly on needles and sharps manipulation. PMID- 18409880 TI - [Technical survey of safety levels established in building sites in the Pescara area]. AB - In 2006, in the context of the initiative aimed at "Promotion of safety and health of workers in construction yards", the Operative Unit of Occupational Health, University G. d'Annunzio, in collaboration with the Construction School of the Province of Pescara and INAIL, started a survey of construction yards around Pescara aimed at collecting data regarding respect of the specific law concerning construction yards and the risk perception of workers. The survey of 100 construction yards provided useful information towards a concise picture of safety levels obtained. In addition 300 interviews were carried out with workers to find out about their knowledge of accident prevention, how important they considered the use of personal protection devices, how often they used them, reasons for not using them, the devices most used,....and any injuries that occurred. The studied showed that, although the situation as improved compared to the past, further steps need to be taken. PMID- 18409881 TI - [Malignant pleuric mesothelioma in Sicily. Epidemiologic observations during the time 1998-2005]. AB - Pleural malign mesothelioma is the only one to which we can straight attribute to an exposition, in professional and ambient ambit, to a good identified agent and, precisely, to the fibres of asbestos. Objective of our work has been to verify the incidence of the pleural malign mesothelioma in Sicily and in particular in the Messina's area, during the period 1988-2005. We have examined the epidemiologic data of the period 1988-1997 and 1998-2005 through the consultation of the Mesothelioma Sicilian Register; Mesothelioma Italian Register and of the available documentation in the Health Superior Institute. The study showed that the pleural malign mesothelioma in sicily and in Messina's area is reduced for the elimination of the asbestos and for the low use of asbestos in the sicilian industry. PMID- 18409882 TI - [Pleural mesothelioma in barman with probable occupational exposure to asbestos]. AB - The case of a barman who suffer from Malignant Mesothelioma (MM) has been signaled. For this case it has been documented a possible source of occupational exposure to asbestos caused by the presence of asbestos components in the professional espressos coffee machines. In some gaskets which are part of these coffee machines, we verified the presence of chrysotile fibers. Italian National Mesothelioma Register have reported a MM case with a professional origin arised in a barman with a certain diagnosis in 1999 (Tuscany Region). PMID- 18409883 TI - [Health surveillance in workers with a previous exposure to asbestos: a method of retrospective evaluation of exposition]. AB - The medical surveillance of the previously exposed to asbestos like method of retrospective appraisal of the exposure. The medical surveillance of the previously exposed to asbestos is effected on indication of the Competent Physician by the DL 257/2006. An aspect that often countersigns such typology of surveillance is the lack of relative data about past environmental asbestos exposure. There've been submitted to sanitary controls 140 subject employees in a steel metal company where in past activity of heavy carpentry has been developed. All the subjects have been submitted in the period 1998-2007 to medical visit, PFR with DLCO, radiography of the chest, in some cases TC and BAL. The past exposure has been resulted inclusive between 1962 and 1981. None of these workers has developed asbestosis, while the 10% of the subjects have showed bilateral pleural plaques. 12 subjects have been submitted to close examination through BAL for the determination of the internal dose of asbestos with comparison of middle values of 3.9 ca /ml. Four cases of pulmonary neoplasm and any case of mesothelioma have been diagnosed. This experience shows as the medical surveillance of a homogeneous group of workers can furnish useful data also to frame previous asbestos exposure in absence of environmental data. PMID- 18409884 TI - [Activity of the Campania Register of Mesothelioma from July 2003 to October 2007]. AB - Campania Mesothelioma Register was established in 2002; its purpose is to record every case of malignant mesothelioma that occurs in the registered population of Campania. Its aim is to identify new dangerous asbestos sources, by giving patients a questionnaire about their working and living habits. The questionnaire used is by National Mesothelioma Register (ReNaM). Analyzing carefully the answers to the questionnaire, it is possible to classify patients' exposure with a code given by ReNaM. By means of a recognition identification network, COR Campania identified 492 cases of malignant mesothelioma (pleura, pericardium, peritoneum and tunica vaginalis of the testis) diagnosed between 1996-2007. The analysis of the ReNaM questionnaire confirms a prevalence of professional exposures (71%), however unknown exposures (15%) also play an important role. The economic sector that determined the greatest number of professional exposures are Construction (17.5%), Industrial Metalwork (13.13%), Railway Car (9.3%), Vehicle production and maintenance (8.16%), Ship building (7.5%). PMID- 18409885 TI - [Case studies on the malignant mesothelioma in Pellezzano (SA)]. AB - 41 malignant mesothelioma cases were reported between January 2000 and April 2007 in the province of Salerno. The small town of Pellezzano, near Salerno, has more cases than any other urban centre in the province; five mesothelioma cases (three male and two female) in Pellezzano (population 9,171 in 1991) means a standardized incidence rate of 32.7 males and 21.8 females per 100.000 inhabitants. That's very alarming, considering that in Italy mesothelioma standardized incidence rate per 100.000 inhabitants is 2,98 for males and 0,98 for females. Campania Mesothelioma Register aims to investigate which kind of exposure caused this abnormal incidence rate. All five patients answered the questionnaire and a team of doctors performed an on-the-spot investigation. The studies verified the existence of two kind of asbestos exposure, professional (Cotton Manufacture, Construction, Foundry) and environmental (cement-asbestos pre-fabricated since 1980 earthquake), that have to be analyzed further. PMID- 18409886 TI - [Evaluation of plans for reclamation from products containing asbestos to ensure safety to the workers and the general population]. AB - Fifty-two working plans for abatement of asbestos-containing materials (ACM) were examined being presented over 12 months at a Public Occupational Health Service in Apulia (Italy) by authorized companies. Most of the interventions took place in urban areas and civil buildings and lasted few days with employment of a limited number of workers. They mainly consisted in removal practices. The weight of ACM abated varied from 30 to over 14,000 kg. Air monitoring of asbestos fibres was included not in all plans. When airborne fibres were detectable, the threshold of environmental pollution (20ff/L) was sometimes reached or exceeded. This finding appeared to be relevant for asbestos exposure of workers and general population. A higher fibre dispersion occurred during abatement of amounts of ACM more than 1,000 kg. The Occupational Health Service should be legally and operatively able to make regular workplace inspections, especially when large quantities of ACM have to be abated. PMID- 18409887 TI - [Subjective perception and comfort/discomfort ergonomic evaluation in school environment]. AB - In order to carry out the present study a school place representative of the entire Italian school have been identified, an Institute within Rome province have been selected. The first step has been the measurement of the dimensional and structural features of the classrooms considered, their furniture and work posts, also considering the activities carried on in the classrooms and their environmental conditions. Pictures and videos have been shot so as to have a deep analysis of the ergonomic level of the work-posts and, at the same time, an inquiry to understand the needs of the students with respect to the environment "classroom", the work-posts and didactical tools utilized, by means of a multiple answers questionnaire, drawing, and the implementation of the shared design method. Applying with the young users the creative sharing design method it have been produced the "diagnosis paper", where the problems and the criticalities relevant to the well-being and healthiness of the users of the school environment have been listed, and which allow to identify patterns, useful basis for future re-designs. The data obtained have been then the subject of an exact and deep analytical and statistical examination carried on by the multi-discipline. PMID- 18409888 TI - [Organization of work, hardship at work, mobbing: new problem areas that must be considered by the competent doctor in formulating fitness evaluation]. AB - In recent years, in addition to the more common occupational-health risks (chemical, physical, biological), increasing importance is given to psychological and psychosomatic illness as a result of malfunction in the organization of work: burn-out, mobbing, maladjustment at work. In order to be able to evaluate the influence of such phenomenon whilst judging fitness in some working realities in Sicily, 18 small, medium and large dimension companies with a total of 1413 employees were examined. The typology of the work embraced the industrial, health, information-technology, construction and telephony sectors. The research involved analyzing, with the respective competent physicians, (in a few cases the same physician dealt with more than one company) the Risk Evaluation document, the clinical and risk records and the fitness evaluations. The analysis of the data obtained pointed out that of the 1413 employees observed, 87% was judged fit, 2% fit with limitation and/or prescription, almost 1% not fit. Notwithstanding the type of judgment, it was ascertained how in the above evaluation, only classical "physical" risks were evaluated (noise, MMC, VDT, etc.). The worker is judged as an individual removed from the general context of the company, of it's organization, or, as one who participates only by means of his various organs and apparatus, and who can singularly become ill because of exposure to some occupational risk. As a matter of fact, in none of the DVR's of the 18 companies object of the research, was an element of evaluation of the working organization found. Nor on the communication means, on the transparency of the procedures, on the quality of relations, lack of instruments or physical sites fit to working activity, nor to emotionally extreme working activities. It must therefore be emphasized that the competent physician only possesses the individual pathologies encountered in the various working situations. He lacks elements of evaluation that would supply useful information on the working organization, and on the effects the latter can have on the health of the employees, and, therefore on their fitness to work. The latter; in our opinion, is data that cannot be disregarded in the correct judgment of psycho-physical fitness, as foreseen by the regulations in force. PMID- 18409889 TI - [Assaults and nuisances in health care environment]. AB - A descriptive exploratory survey was used to investigate the occurrence of physical workplace violence among health care workers as well as the complaints and responses to such violence. The questionnaire was used for collecting data from a sample of 355 workers. Results showed that 25.6% of hospital workers reported physical aggression at any time of the working life, 8.5% in the last year. Injured workers showed significantly higher environmental complaint score than other workers. The results of the study indicated the importance of adopting hospital policies for dealing with incidents of violence. PMID- 18409890 TI - [Job satisfaction. Use of the Job Satisfaction Scale (JSS)]. AB - Job satisfaction is an important determinant of wellbeing and a moderator of stress at work. The Warr's Job Satisfaction Scale (JSS) is probably the most used questionnaire to assess job satisfaction. The aim of this paper is to evaluate the Italian version of the JSS. The questionnaire has been distributed to 632 health care workers. Results show that the Italian version of JSS has good reliability (Cronbach's alpha = 0.94). Principal component analysis revealed that a significant percentage of the variance (52%) was explained by a single factor which included all the 16 items. Varimax orthogonal rotation yielded the same two factors observed in the original questionnaire: intrinsic and extrinsic job satisfaction. The JSS may be an useful tool in the assessment of psychosocial risk at work. PMID- 18409891 TI - [Participatory risk evaluation. The role of the surveillance]. AB - Workplace assessment is a critical issue in small firms, and even in bigger ones if workers are scattered in large working areas. The traditional "top-down" approach, based on centralized risk assessment and hardly understandable documents, may have limited impact on workers' safety behaviour. The Local Sanitary Unit RMF tried to improve workers' participation to risk assessment, through auditing techniques. Waste workers signalled occupational condition of biological and chemical risk, which had not been taken into account in the Corporate Risk Assessment Document. The participatory approach to workplace risk assessment proved to be an easy and not expensive method to increase workers' awareness of risk and compliance to hygienic measures. PMID- 18409892 TI - [Influence of work climate on the sick building syndrome]. AB - The potential risk factors for sick building syndrome (SBS) are not yet well elucidated. A questionnaire was administered concerning environmental conditions at the work place and complaints before and after the take-over of one company from another corporation. Workers had to move into new company, but were still working in old office building. Before the change, environmental objective and subjective conditions and workers' wellbeing were within normal range. The change did not involved air quality, however results of the questionnaire demonstrated an increased irritation of the mucous membranes and a reduction of well-being. The level and severity of symptoms appeared to be related to perceived indoor environment quality, and depression score. High stress related to work changes was found to be significantly associated with SBS symptoms. The psychosocial work environment can be an important predictor of SBS symptoms. PMID- 18409893 TI - [Proposal for an evaluation method of the psyco-social risks (stress) and for the orientation of the sanitary surveillance]. AB - The interest for the problematics linked to the stress in work environments has been till now limited to the consideration of its effects on health, even if the D.Lgs 626/94 obliges the employer to evaluate also the psyco-social risk, as has confirmed a sentence of the European Court of Justice. The present survey, lacking valid instruments to be found in literature, aims to experiment and evaluate a principle for the identification of causes, thus creating a model for the evaluation of risk also according to the indications published in the Document for the Consent of SIMLII in 2005, which can be used by Prevention and Protection Services and by Competent Medical Doctors. In the area of risks evaluation and of the attainment deriving from them. The model of evaluation of the risk deriving from work organization (stress), object of the present survey, has been sperimented in a sample composed of 268 employees in 13 municipal administration belonging to biographically known categories for stress risk afferent to 23 homogenuous organizational structures (traffic officers and nursery school teachers). The valued risk has been introduced in the VDR document and the indications for the sanitary surveillance have been formulated. PMID- 18409894 TI - [WOA: a questionnaire for the evaluation of work organization. Translation and validation of the Italian version]. AB - The Work Organisation Assessment questionnaire (WOAq) has been translated into Italian, back-translated, and modified. Data were collected from 160 health care workers and 50 white-collar bank employees. Employee wellbeing was assessed by the General Health Questionnaire (GHQ12), while work stress was assessed by the Karasek's Job Content Questionnaire (JCQ). Reliability, as measured by Cronbach's alpha, was very good (0.95). Principal component analysis revealed that a significant percentage of the variance (41.8%) was explained by a single summative factor which included 25 of the 28 items. Varimax orthogonal rotation yielded the same five factors observed in the original questionnaire (reward and recognition, quality of relationship with management, quality of physical environment, quality of relationship with colleagues, workload). The WOAQ sum score was positively related to social support, as measured by the Karasek's JCQ (Spearman's rho = 0.523; p < 0.001). It was negatively related both to job strain (Spearman's rho = -0.516; p < 0.001) and psychological distress GHQ (Spearman's rho = -0.365; p < 0.001). In conclusion, the Italian version of WOA maintains the original characteristics. The questionnaire has strong association with job stress and employee wellbeing, and it may be useful in risk assessment procedures. PMID- 18409895 TI - [Stalking in work environment]. AB - Stalking is a form of interpersonal victimization that can have irrevocable effects on the lives of victims but is frequently misunderstood and minimized. This article presents a case study of a woman who was stalked in the workplace by a 39-year-old male colleague, and offers recommendations for occupational health clinicians charged of medical surveillance of workers (so-called "Competent Physicians") who have to cope with stalking at the workplace. The prevalence of stalking provides many opportunities for Competent Physicians to intervene, but first they must recognize and understand the problem. The study underlines the inadequacy of the current legal and medical responses to the needs of these victims. PMID- 18409896 TI - [Two tools for health surveillance of job stress: the Karasek Job Content Questionnaire and the Siegrist Effort Reward Imbalance Questionnaire]. AB - Occupational stress is currently studied by the Job Demand/Control model of Karasek, and the Effort/Reward Imbalance model of Siegrist. In this study we have translated into Italian and validated the short form of the Job Content Questionnaire (JCQ) and of the Effort Reward Imbalance Questionnaire (ERI). The questionnaires were applied to 531 health care workers during periodical medical examinations. Estimations of internal consistency, based on the correlation among the variables comprising the set (Cronbach's alpha), in each case were satisfactory (alpha ranging from 0.76 to 0.89), with the exception of the control" scale of JCQ (alpha = 0.57). Exploratory factor analysis showed that "control" scale of JCQ, and "reward" scale of ERI could be divided into two and, respectively, three sub-scales. The Karasek's and Siegrist's models made distinct contributions to explaining perceived work stress. Both JCQ and ERI questionnaire may be useful in occupational health. PMID- 18409897 TI - [Anxiety and depression at work. the A/D Goldberg Questionnaire]. AB - The Italian version of a short anxiety and depression questionnaire developed by Goldberg et al. (1988) was applied to 1918 health care workers during periodical medical surveillance at work. Internal consistency reliability coefficient (Cronbach's alpha) value was 0.82 for anxiety scale (A), and 0.78 for depression scale (D); both scales can be retained "good". The factor structure of the two scales was examined through use of the factor analysis technique. The anxiety scale showed an homogenous pattern: a significant percentage of the variance (41.5%) was explained by a single summative factor which included all the items. Varimax orthogonal rotation of the D-scale variables yielded two factors, both typical of depression: reduced activity (6 items, 28% of variance), and poor self image (3 items, 22% of variance). The scales would be used by occupational health physicians in medical surveillance as indicators of stress levels. PMID- 18409898 TI - [Analysis of causes of occupational stress with the QFSL, questionnaire on occupational stress factors]. AB - Stressors at work are classified, according to Kalimo (1980) and Cooper (1976) in six broad categories: role within organisation, intrinsic stressors, interpersonal problems, work climate, career, social context. The 40-item Factors of Stress Questionnaire has been proposed as a screening instrument for causes of stress at work. The factor structure of the questionnaire was evaluated in 371 health care workers. Internal reliability analysis was also conducted. The Factor Analysis, according to the Kaiser rule to drop all components with eigenvalues under 1.0, gave nine principal components, with a 58% cumulative variance explained. However, most of these components had poor comprehensibility. The first component was significantly related to most of the items. The questionnaire showed good reliability (Cronbach's alpha = 0.93). PMID- 18409899 TI - [Workers risky for others on the Web]. AB - Hazardous workers (HWs) are they who, depending upon their pathologic conditions, ageing or addictive behaviour, may pose at risk the safety and health of their colleagues, customers, and other people. Physicians charged of the medical surveillance of workers (Competent Physicians, CPs) are called to assess fitting for work of HWs. The aim of this paper was to analyze the frequency of debate about HWs in the online forums of the Italian CPs' website. Results show that there is growing attention on the HWs issue. Different stakeholders are involved, and some ethic dilemma are posed. A systematic approach to the problem, and the formulation of guidelines for CPs, is highly desirable. PMID- 18409900 TI - [The relationship between mobbing and depression syndrome in the female working population of service industry: the problem statement and the prevention strategies in Sicilian environment]. AB - The concept of "work" and the social interactions in the workplace contribute to develop the working satisfaction and the social and personal identity of the adult. The aim of the survey is that of checking up on the presence of a statistically significant relation between The Depression Syndrome and The Mobbing Phenomenon. It is necessary to understand and value if and how some of the employer's behaviours could have a heavy influence on the worker's psychophysical condition, up to causing a state of anxiety and depression. It has been used the now famous "LIPT" (Leymann Inventory of Psychological Terror), elaborated by Leymann at the beginning of the 90's. It is a short anonymous questionnaire recognized all over Europe as a standard to confirm the Mobbing existence in organizational contexts. The current version of the questionnaire is called "LIPT EGE", a more suitable tool to check the seven parameters of the Mobbing determination, as well as the quantification of the consequent harm. Some scales for depressive symptoms evaluation have been added to the Mobbing questionnaire: The Self Rating Depression Scale (SDS) and The Depression Status Inventory (DSI) of Zung. The questionnaires are administered to 500 workers women in public and private corporation in Sicily, exactly in Siracusa and Catania provinces. 206 women aged 34-50 have answered the questionnaires. In addition to the age and the sex, the distinction variables of the examined people are the position in the firm, nationality, vocational qualification, and the yearly gross income. The research has begun in july and has continued on august 2006. Attention has been focused only on the administrative sectors. Through the chi2 test and the exact Fisher test, the dependence between the changeable presence of depression in the two questionnaires (Zung 1 and Zung 2) and every single variable of the questionnaire on the Mobbing has been pointed out. In particular, the results establish a connection between the presence of depression and systematic isolation, and depression and the changes in the working tasks. The survey has pointed out the working conditions that may lead to Mobbing situations and to psychological frustration; it follows that the Mobbing prevention is a key element if you want to better working life and avoid social emargination. PMID- 18409901 TI - [Evalutation of neoplasms prevalence as a consequence of stressful events in the occupational and non-occupational field]. AB - Starting from the hypothesis that stressful events, badly elaborated by individuals, can bring a decreasing activity of immunitary system and represent a co-factor in development of breast cancer, analyzing which events are more stressful and as a consequence, which are the ones causing a decreasing of immunitary defence. This study considers the breast cancer in large increasing amongst women. 60 women compiled the Events of Life Questionnaire (Qav) and they have been divided into two groups: 30 elements affected by breast cancer (experimental group) and 30 without evident pathologies (control group). Their age was included between the 25 and 75 years old, they came from the same social class, cultural extraction, instruction level. Throughout every single interview, each one lasting about an a hour with hospitalized patients, we potentially find a depressive personality. The statistic analysis gives extremely interesting results regarding negative events of the affective life: underlining a strong dependence among negative events of affective life relating to the development of the breast cancer This interest start from the evident limit in the medical practice, considering organic problems as the only for disease development, excluding emotional and psychological factors which are part of human being's life. Anyway, today is possible to find especially in onchological medicine, the consciousness that the medical or surgical intervention is not the only duty but is also important to underline, in a more complete view, that the patient's care also depends from the ability of questioning about patient's conditions in every day life; as shown in this study which underlines the necessity of psychotherapeutic intervention for patients affected by breast cancer, specially in pre and post operating phases. PMID- 18409902 TI - [Evaluation of the psychological well-being and coping strategies in a population of ex asbestos exposed on its work place]. AB - The aim of the current study was mainly to asses the coping strategies used by a sample of subjects (n = 100) and its psychological well-being. The INAIL had recognized these subjects in the past time had been massively exposed to asbestos in their work place. An anonymous self-report questionnaire, made up of (a) an explorative section, (b) the Carver's COPE, (c) the Carol Ryff's Psychological Wellbeing Scales (PWBS), has been used. Both tests parametric (Chi Square and t Student analysis) and not parametric (Mann-Whitney Test) were carried out in order to assure strength results. The survey data indicated that there were an acknowledge about the asbestos-related diseases' action and a general use of "avoidance" coping strategies. Contrary to expectation concerning the PWBS, there was a statistically significant higher score in the "exposed group" than the "control group" (n = 50) as regards the "Positive Relations with Others" and the "Personal Growth" scales. Different variables may concur to explain the results of this study. PMID- 18409903 TI - [Anxiety, job stress and job insecurity among teachers with indefinite or definite time contract]. AB - Object of this study was the occupational stress of 336 teachers (276 women and 60 men) with stable or temporary employment in schools of Pescara, town of Central Italy. The levels of anxiety were determined by STAI and STAI II, those of job strain", "job insecurity" and social support by the Karasek's questionnaire, and the perception of subjective symptoms by a 12 item test. There were no significant differences depending on the type of school. The women with temporary contract showed higher levels of "job insecurity", while the men with temporary job showed also higher values of STAI I and STAI II. The scores of anxiety of the women were positively correlated with "job strain", "job insecurity" and perception of subjective symptoms and negatively with social support, while the only correlations of STAI I and STAI II of men showing statistical significance of men was that with "job insecurity. Job strain was negatively correlated with the perception of symptoms both in women and men. These results evidence differences in the occupational stress of men and women; in particular, job insecurity may enhance anxiety in men. PMID- 18409904 TI - [Stress and work. Result of an epidemiological investigation among the voluntary postal and telecommunications personnel of the Calabria region]. AB - The Stress, in consequence of the alterations neuroendocryne to it correlated, as cause of pathology is often suitable, professional and not, among the various. But the circumstance that it can recognize causal moments in the different existential situations makes difficult to bring back the cause of a lot of pathologies to the only working stress somehow in relationship with the working activity. To overcome this "enpasse" I diagnose the authors they apply, in one epidemiological job of theirs, with methodological iter that considers, above all, the efficient and conclusive role of the working factor. PMID- 18409905 TI - [Burnout evaluation in public employees in the South of Italy]. AB - The aim of our study is to evaluate Burnout syndrome in workers of a city in Southern Italy. The study was conducted on a sample of 1779 subjects including (562 teachers, 476 care worker, 549 employees) operating from birth and residents in the province of Messina. The data and clinic information were taken during the health surveillance. All subjects were administered during the health surveillance testing Maslach Burnout Inventory (M.B.I.). The category of Teachers presents 114 (20%) subjects with the syndrome. The category of care worker 74 cases (15.5%), the category of employees presents 83 cases (15.1%). It can be said that today the class teacher is more affected than the others as witnessed by the scientific literature. PMID- 18409906 TI - [Emergency, subjectivity, stress: on the side of the helper]. AB - Rescue and emergency teams are affected by different stress factors. We investigated 201 subjects from Red Cross Organization, firemen "regular" and volunteer, policemen, and health rescue team. Mental fatigue and psychological impact are more common critical factors. 55% of subjects show burnout symptoms, more among Red Cross volunteers, and firemen. Among policemen the emotional exaustion has more pathologic results; depersonalization among fireman and personal accomplishment among Red Cross and firemen volunteers. The normality of the data from other psychophysical health investigation confirm occupational origin of the complaints. PMID- 18409908 TI - [Assessment of occupational stress among workers in rehabilitation medicine]. AB - Emerging evidence confirms the risk of distress in staff involved in the Helping Professions resulting in the possibility of reduced efficacy and efficiency of the structure which offers health services. The aim of the present study was to reveal the presence of burnout and its relationships with environmental and organizational factors of health care professionals enrolled in medical rehabilitation. 88 health care professionals underwent a psychometric assessment by filling the Organizational Checkup System Questionnaire (OCS) for measuring burnout and associated organizational factors. Data analysis demonstrated the presence of major emotional distress (p = 0.05) in the group who had an executive role (social-assistants and assistant nurses), especially regarding organization factors linked with: Overwork (p = 0.009), lack of Leadership (p = 0.01), lack of Comprehension of modifications offered by the structure (p = 0.0001) and Professional Development (p = 0.016). OCS revealed to be a sensitive psychometric instrument, especially for the identification of organizational weaknesses, which can stimulate the application of strategies to prevent the burnout syndrome and to improve coping skills. PMID- 18409907 TI - [Satisfaction and stress if the health care system: comparison of two different situations]. AB - We examined two different health care settings to identify if some stress and satisfaction factors are common. In both neonatological ward and psychiatric residence the relation with the patient has been perceived as gratifying but in paediatric setting it is perceived through procedures and protocols, on the opposite site, in the psychiatric setting the daily life care is perceived as a low technical degree. Formation of workers is indispensable tools to understand the "technical" aspects of the work, without promoving a remoteness from the patient. PMID- 18409909 TI - [Management of occupational stress among patients with cardiac diseases]. AB - The object of our study was the evaluation of psycophysic condition, stress perception and coping in a sample of subject in cardiac rehabilitation. Our study recruited 48 worker patients (44 male, M = 50.5 +/- 8.8; 4 female, M = 50.8 +/- 8.7). They were submitted to: 1) cardiac evaluation and rehabilitation in DH regimen; 2) psychological assessment; 3) work's characteristic evaluation. The evaluation were made at the moment of recruitment and six month later 42 subjects went back to work after 81.8 +/- 49 days; 2 subjects had a worsening of their clinic conditions not consistent with an occupational resumption; 4 subjects didn't go back to work. In the 42 worker subjects we have find a significant decrease of anxiety (p < 0.019) and depression levels (p < 0.004); a significant improvement of Quality of Life perception in its physical (p < 0.000) and psychic aspects (p < 0.021) and an improvement about the ability to reorganize the work to better menage stress (p < 0.012). PMID- 18409910 TI - [Follow up of person-related bullying and work-related bullying: evaluation of effectiveness of interventions]. AB - Person-related bullying is regarded as a form of stress capable of cause negative effects on workers' health, potentially leading to psycho-physical symptoms, alterations of mood and personality, psychiatric disorders such as anxiety depression disorder, chronic adjustment disorder and posttraumatic stress disorder. At the Centre for the study of psychosocial risks and pathologies at work of Verona the health state of victims of person-related bullying and work related bullying and their working conditions are periodically evaluated. On a regular bases these workers receive psychological support and, if necessary, psychiatric treatment; at the same time interventions on working environment are undertaken when possible. Verification of effectiveness is then carried out by means of interviews starting six months after the first consultation. Two questionnaires are used; a specific one for the assessment of working environment and health conditions, and Goldberg's General Health Questionnaire--G.H.Q-12. Improvement of health state related to better working conditions was obtained in general. PMID- 18409911 TI - [A follow-up study to assess stress and burnout in healthcare workers. Preliminary data]. AB - To compare data obtained in a pilot study (2002) which evaluated the risk from exposure to stress and burnout in health care workers, the same three subjective questionnaires were administered: the Job Content Questionnaire (JCQ), the State Trait Anxiety Inventory (STAI), the Maslach Burnout Inventory (MBI). The evaluation considered 294 workers employed in eight units: two of the six previous units were reorganized in four subunits. Preliminary data showed that questionnaires' scores have a concordant trend in the different units, as observed in 2002, confirming the validity of the instruments adopted. Results indicated a decreased level of the perceived stress in all the units, especially in the Third Division. We can hypothesize that the new organization, workers' turnover during the 5 years and the effect due to individual variables could have contribute to the observed variation. No significant associations, compared to 2002, between questionnaires' scores and task were found. Further evaluations, including measurement of objective parameters, will be carried out to complete the follow-up study and to determine which variables could have a role in the variation of the levels of stress and burnout's subjective perception. PMID- 18409912 TI - [Occupational risk stress in shift workers]. AB - Aim of our study was to assess the stress risk at work in a group of nurses. 197 workers (nurses, auxiliary, physiotherapists, midwives and technicians of laboratory) were enrolled divided in two sets respectively of 100 people (subset A) with daily and nightly shifts and 97 (subset B) only daily workers. The mean age was 43.29 years (DS +/- 8.13) in subset A and 46.85 years (DS +/- 7.48) in subset B. For the survey S-90, a questionnaire composed by 90 items faces to frame some of the possible factors of risk has been used and the relative effects on health's state induced by the occupational stress were analyzed. The results show a possible greater risk of daily sleepiness in workers with nightly job. The majority of hypertension workers in subgroup B could depend more on the elevated mean age of this sample. Although a greater perception of the psychological load has been found, statistically meaningful association between the insurgence of gastrointestinal symptoms and shift was not showed. PMID- 18409913 TI - [Theory models of burnout: aspects of risk assessment and preventives measures in health care operators]. AB - The figures appointed to prevention is to assess all the risks and working notes that the more complex when you take into consideration the risks related to the organization of work, especially in certain types of work in which a proper assessment can not ignore the intrinsic problems of the job done. The studies that have been employed in particular health professions speak of a phenomenon of job stress response called "Syndrome Burnout." Our study includes a description of current research, in literature, made on the issue and particularly on the theoretical models of Burnout (Models of competence / effectiveness--psychosocial Models--phases of Burnout); Below is sought to draw in some health care workers, "the individual characteristics" of the subjects that I am concerned, identify the causes and effects, outline proposals for action for risk assessment, defining prevention measures, for the early identification of symptoms, acting on variables that affect the occurrence of the syndrome. PMID- 18409914 TI - [The MMPI-2 test in the evaluation of the mobbing syndrome]. AB - MMPI-2 test is widely used in psychodiagnostic evaluation as well as in the evaluation of psychic pathologies related to psychosocial adversative events in working activities. A more efficacious use of MMPI-2 test in the working context can be achieved by the individuation of indexes able to facilitate the evaluation of mobbing syndrome. This work is based on the analysis of 150 cases (39.7% women and 60.7% men, 30 to 60 years of age), evaluated through an accurate examination of working history and a series of clinic conversations, followed by psychodiagnostic evaluation. The average of T-scores on the clinical scale, the content scale, the PK addition scale and the validation scale were calculated from tests. Pathologic high T-scores on the scales Hs, D. Hy e Pa were found in subjects having positive mobbing anamnesis. These results strongly indicate the presence of clinical specificities in workers exposed to working harassments, and that these specificities can be efficaciously evidenced by the MMPI-2 test. PMID- 18409915 TI - [Subjective perception of maladjustment risk factors]. AB - Maladjustment at work results from organizational and relational features of the work, the so-called fourth type factors; they include working hours, ways and contents of working activities, and horizontal and vertical business relations. The study reports the percentage of sensed disturbing factors in workers with maladjustment and disaffection at work. Data have been taken from 1382 white collars, 1117 males and 265 females, observed from January 2006 to June 2007 for Health Surveillance. Maladjustment prevalence was higher in females than in males. As individual variables, ageing and family care increased the prevalence of maladjustment among females, whilst a higher prevalence of maladjustment were found in youngest and unmarried males. A very different perception of work harmfulness were found between sexes. As risk factors, female have denounced more wear and tear and authoritarian management; male denounced physical strain. PMID- 18409917 TI - [The perception of stress in the European stakeholders]. AB - Stress at work is a thoroughly studied issue, also on the basis of ILO, WHO and European Parliament guidelines. Nevertheless, although there is a great amount of research on causes of occupational stress and on its impact on workers' health, few investigations have been carried out on stress perception and the known data shows that there is no unanimous point of view among the different parties in the world of work (employees, employers, trade unions and institutional bodies). While on the one hand some European stakeholders have a poor perception and recognition of stress risk (ISPESL, 2004), on the other hand one of the occupational health problems most frequently reported by workers is stress (European Foundation, 2005). The PRIMA - EF project (Psychosocial Risk MAnagement - European Framework) has been developed in this context, and it is funded by the Sixth EC Framework Program (FP6). It is carried out by an international consortium with the participation of ISPESL (Department of Occupational Health). The project deals with occupational stress management with a special focus on how such an issue is perceived by the different European stakeholders. PMID- 18409916 TI - [Migration, work flexibility and early retirement of nurses in Italy]. AB - Several work organizational changes have been occurring in the health sector in the last years. Workforce is rapidly changing and particularly nurses. In Italy there is a growing shortage of nurses (5,4/1000 inhabitants versus 9,26 in EU25) due mainly to early retirement and insufficient turnover of nurses coming from educational programmes. Work flexibility and facilitation of migration flow of foreigner nurses have been introduced in Italy to face the problem. The present study is aimed at investigating the impact of such a change in the nursing sector in Italy and its possible impact on occupational safety and health. In view of this, the main institutional and category Sources were used. According to IPASVI estimates, the shortage of nurses in Italy is 99,000. Out of a total of 342,000 active professional nurses, 20,000 are foreign, mainly from European countries. Nurses are specially affected by accidents at work, mainly musculoskeletal disorders. The study shows a higher vulnerability of migrated nurses and nurses hired through cooperatives. Therefore urgent measures are needed to promote their integration and assess educational and training needs in the field of health and safety at work. PMID- 18409918 TI - [Seasonal atypical jobs in the sector of the floriculture in Calabria--prevention of the risks and safety on the job]. AB - The search concerns over three hundred precarious workers in the sector of the floriculture of the region. They has been submitted to medical visit before the assumption. The workers that have had other experiences of job in the same productive sector; have reported to never have effected periodic sanitary controls. The medical visit is the occasion to inform the operators on the specific working risks and on the correct procedures of job. Some workers have probably reported some consequent troubles to phyto-drugs poisonings. These poisonings are not been signalled to the competent authority for fear not to be recalled to the job. The medical visit before the assumption establishes a relationship of dialogue and trust that is at the base of every correct and effective intervention for the prevention and the tutorship of the health of the workers. PMID- 18409919 TI - [Building of a method for the assessment of work related risks at the ULSS n.1 company of Belluno]. AB - This paper describes an experimental method of risk assessment in a public hospital. A uniform method of danger checking out, valid for all hospital units was developed, and a objective and standardized system of occupational risk assessment was implemented. In fact, the quantification of risk is expressed in a numerical code that allows computerization of the collected data. The method is based on check list for the danger detection, on worker's data sheet, on criteria for risk factors encoding and for risk quantification. Vantage of this system is applicability into all the hospital units, even in presence of different occupational risks. Another vantage is the involvement of staff in the management of work safety. PMID- 18409920 TI - [Microclimate problems linked to multisplit air conditioning in large offices]. PMID- 18409921 TI - [Evaluation of gasoline exposure during the tankers loading in two storage gas plants]. PMID- 18409922 TI - [Exposure to cancerogenic agents at work. Tools for their management and correct identification in relation to the type of work]. PMID- 18409923 TI - [Risk perception in a group of dentists from Abruzzo]. AB - Dentist's office represents an area with high infectious risk both for workers and patients. With our study we tried to analyse the relationship between dentists and prevention. We studied 104 dentists who had to answer to a questionnaire composed by 25 questions on topics such as job management, risk perception, measures adopted for prevention and knowledge about normatives regarding safety at work. From the answers we can conclude that dentists know the risk present in their work and use correctly the individual preventive dispositives; make blood control to test hepatitis markers, in particular HCV. But only 68% made HBV vaccination. Many dentists,even if they know the way of transmission of HIV, avoid to cure positive patient. PMID- 18409924 TI - [Noise related risk: assessment of the efficacy of the prevention in the light of the new legislation]. AB - Italian law on occupational exposure to noise identify 87 dBA as ceiling exposure level and 80 dBA as action level; the respect of ceiling level can be achieved by using devices, as ear plugs, whose efficacy must be verified. We studied 8119 audiometric of workers exposed to noise between 80 and 87 dBA. The hearing level of those who use ear plugs is significantly better than who don't use. Different devices show different efficacy, but for all, in the exposure setting studied, the level of protection that is provided is enough safe for othological protection. The workers must be informed about the efficacy of the protection and sensibilized to the use of the ear plugs within the periodical clinical surveillance. PMID- 18409925 TI - [Measurement of energy expenditure in a group of construction workers during work]. AB - The aim of this study is to show the results obtained from measuring energy expenditure (EE) during work in a group of male construction workers through a new portable device of practical use and low cost. The instrument consists of an electronic bracelet multisensor system for measuring EE, which is called Armband Sensewear Pro2 and is produced by Body Media. The disposal has 5 steel hypoallergenic sensors which detect the skin and the near body temperature, the environment temperature, the electrical skin conductance, and an internal two axes accelerometer which detects the transverse and longitudinal acceleration. The analysis of these parameters through software allows you to derive the individual EE. The study describes the results obtained by applying Armband to a group of 10 construction workers performing the following jobs: bricklayer, tiler, crane driver and "rock climber". The results allow to assess and confirm the high EE related to the work of the subjects enrolled in the study. In conclusion the authors argue that Armband, already validated experimentally in several areas of Medicine, can be a useful tool available to the Occupational Physician, in order to determine the EE for workers who perform heavy tasks, such as those that exist in the construction industry, which are characterised by high levels of EE and poor reproducibility of tasks. Moreover, Armband appears to be an useful tool in connection with the dual possibility on the one hand to assess fatigue related to certain skills and jobs, and on the other side to contribute to the metabolic evaluation of cardiac patients during rehabilitation at work. PMID- 18409926 TI - [Occupational hazards in management of an experimental constructed wetlands]. AB - The sewage from urban settlements is an important cause of water pollution. Recent Italian legislation offers new possibilities for intervention in the purification of wastewater. The "Decreto Legislativo" 152 of April 3, 2006, indicating "Environmental Standards", recommends the possible use of constructed wetlands considered particularly suitable for small and medium communities according to low operating costs, lack of energy, and manageability. The Authors, after describing the characteristics of these facility, analyze the occupational risks associated with the control and maintenance of a experimental subsurface constructed wetland (H-SSF, SubSurface Horizontal Flow) coming to the conclusion that the management of this type of facility has a low risk of exposure to biological agents. PMID- 18409927 TI - [The perception of risk in construction workers]. AB - Accidents in building sites led us to study risk perception in workers employed in this field. We disposed of 300 workers, aged 18-65, who had to answer to a questionnaries were signing among seven risks the more present in their activity, chosing among five levels. So for each risk the worker had to quantify parameters such as: danger, frequence of accidents, actitude to risk-control, training on risk. The risk of falling from high sites has been perceived as extremely dangerous but it is possible to control it. Less dangerous is eyes exposition to chemical agents or dust. Manual movementation of weight and lacerations would be the most frequent source of accidents. Moreover would be insufficient the capacity to control the rick related to Weight manual movementation and vibrations. Workers answered in questionnaries that risk training on vibration was not sufficient, on the contrary they knew the risk of falling from high site. So we conclude that to reduce accidents in building sites it is necessary make better training on field, modifying the individual behavior among workers. PMID- 18409928 TI - [Risk evaluation of contact with pesticides in Naples municipality gardeners]. AB - Pesticide use still now is a remarkable risk for all cultivation operators, including farmers and flower-growing workers. Recently, pesticides use on ornamental plant has been demonstrated to be able to determine a risk extension to general population. Aim of the project was to elaborate and to apply a method for pesticides risk evaluation in ornamental plant cultivations, as to define workers exposure and its possible diffusion to not occupationally exposed populations. Main features of method will be sampling and analysis approach. The first will estimate air dispersion of pesticides in garden centre, skin and garment smarminess in nurserymen and leaf adhered amounts, in concomitance with treatment and periodically to estimate natural decay. The latter will characterize pesticides by means of different methods (liquid and gaseous chromatography, spectrophotometry and spectrometry). Final aim will also be to identify the most reliable analytic method, as concerns accuracy, sensitivity and specificity. PMID- 18409929 TI - [The effectiveness of measurements of chemicals and biological agents in the particulate matter of the workplace: the contribution dell'INAIL-CONTARP]. AB - Quality assurance of measurements is a crucial target for the laboratories interested in improving their analytical performances. To this aim, the Italian Workers' Compensation Authority (INAIL) has undertaken inter- and intra laboratory net in the fields of chemical and biological agents measurements for workplaces risk assessment. Purposes and results of up to date experimental activities are summarized in this work. PMID- 18409930 TI - [Evaluation and risk management of the risk of robbery]. AB - The carried out work have permitted to make a technical evaluation of the risk robbery in a big Company of services and to formulate a strategy in order to prevent the criminal event and its effects on the workers' health. At this point an algorithm of prevision of risk has been formulated, taking account the elements of probability of criminal act and those ones referring to the relating damage according to the following formula: RISK(R) = PROBABILITY(P) x DAMAGE(D). The use of this method has proved useful to obtain the right evaluation risk robbery and it has permitted to elaborate a document of Health Protection to use towards the workers who are involved in the event with the active participation of the Company doctor. PMID- 18409931 TI - [Silicoasbestosis: report of three cases]. AB - Silicoasbestosis is a rare pneumoconiosis due to the combined sclerosing effect of crystalline silica and asbestos fibres. We present here three cases (males, age: 66-75 years), recently reported to the judicial and workers' compensation authorities. The exposures occurred in the building and siderurgic industries, several years before retirement. The radiological pictures were characterized by diffuse nodular opacities, interstitial alterations, emphysema, pleuric plaques and thickenings. The patients described show that, still nowadays, silicoasbestosis may be observed in the clinical setting. Accurate occupational and environmental anamnesis are helpful for the correct diagnosis which, in turn, is important for prognosis and treatment, and in relation to medicolegal issues. PMID- 18409933 TI - [Silica risk in construction industry: an investigation on raw materials]. AB - Typical construction activities like demolition, excavation and sanding can expose workers to silica, but there are few investigations carried out with the aim of evaluating the silica content in raw materials used for these activities (mortars, plasters, cement...). Our intervention has been directed in looking for free crystalline silica in samples of raw materials. We have measured the silica content in these materials comparing declared and real composition found in the products. Our intent was to obtain more information about the working activities which expose workers to free crystalline silica in construction industry and to highlight the silica presence in raw materials also when the companies don't declare it, against labeling rules based on Italian law D.lgs 65/03. PMID- 18409932 TI - [Investigation on health status of silica exposed workers in "cotto Fiorentino" companies]. AB - The manufactured in "cotto" is typical of Chianti Fiorentino with about 400 employees. In 2005. the UF PISLL encountered an exposure to silica more than TLV, particularly for some tasks; were prescribed interventions of prevention and was undertaken an investigation of the occupational health status with occupational health physician. We observed 227 workers, 208 males and 19 females, with average age of 43 years and average age working of 15 years. The habit of smoking tobacco was higher than for the general population. The assessment of exposure to silica has been detected for 59 workers (mean 0.05 mg / mc); at pulmonary function testing resulted: 10 with airway obstruction and 4 airflow limitation; by 140 chest X - ray acquired 1 was interstitial pulmonary disease and 1 was bronchopneumonia. Among the diseases not related to exposure to silica, emerging 42 cases of low back pain, 28 hearing loss, 7 with hypertension. Non uniformity in health surveillance and diagnostic criteria highlights the need to cooperate between occupational doctor in public prevention and control service and qualified occupational doctor to ensure a standard of quality in the prevention of disease in exposed to silica. PMID- 18409934 TI - [Antiblastic drug exposure: statistical analysis of the data coming from environmental monitoring of a site devoted to drug production]. AB - An environmental monitoring strategy was carried out for the determination of surface concentrations of cyclophosphamide (CP), ifosfamide (IF) and 5 fluorouracil (5-FU) in a drug preparation room of an oncology ward. Analytes were determined by wipe tests, liquid-liquid extraction with diatomaceous earths and GC/MSMS or HPLC/UV analysis. The analysed 249 samples showed concentrations of CP, IF and 5-FU varying in the ranges 0.020-18.83, 0.100-26.96, 0.740-208.9 microg/dm2, respectively. It is noteworthy that the 9.3% (CP), 18.6% (IF) and 76.3% (5-FU) of the investigated surfaces showed high contamination levels, with analytes amount higher than 0.5 microg/dm2 and a progressive contamination decrement going from workbenches, floor, hood planes and other examined surfaces (interphone, telephone etc.). A significant correlation (rhos = 0.303, p = 0.001) between the measured analyte concentration and the analyte handled amount was found only in the case of IF, and a diffuse contamination (traces of all the three analytes) was found on all investigated surfaces, even when analytes were not been used during the sampling days. PMID- 18409935 TI - [The pathogens responsible for waterborne infections: the biological risk for agricultural laborer]. AB - The presence of microbiological agents in waters intended for human consumption, irrigation and recreational activity, is a significant risk for the human health. Recent epidemiological data demonstrate an increase of the diseases caused by waterborne pathogens, especially of the gastroenteritis infections, in the industrialized countries. Numerous studies have evidenced the contamination of deep and surface waters; bacteria within contaminated water represent a highly diversified group, that are usually encountered in wastewater and sewage. In particular the pollution of the groundwater is much dangerous, from the viewpoint of drinking water safety. In fact, the groundwaters whenever polluted, have an insufficient self-purification. The aim of this paper was to estimate the presence of waterborne pathogens in groundwater intended for the irrigation. Well water samples were collected at eighteen different agricultural areas located in Calabria and analyzed for the presence of waterborne pathogens. The results shows a diffuse pollution of the wells in all of the monitorated zones. Then out of eighteen examined samples were positive for total coliforms and Enterococcus spp. and the value of total microbial contamination were higher than 100 UFC/ml; eight of these ten wells were contaminated from Salmonella spp. and Shigella spp. (minimal value 2x10(1) UFC/l; maximum value 2x10(3) UFC/l). PMID- 18409936 TI - [The importance of the airborne microorganisms evaluation in the operating rooms: the biological risk for health care workers]. AB - The operating room is a complex environment, traditionally considered at high infectious risk, for both the patients and the health care workers, they can contract diseases, because of the exposure for relatively long times to various dangerous chemical, physical and biological factors. The biological contamination in the operating rooms is mostly imputable to airborne and bloodborne microorganisms, whose primary source represent the staff: patients and operating team, while either secondary sources are the contaminate air introduced from the VCCC system and the use of the infect instruments. About 10% of the hospital infections are determined by airborne bacteria and a variable fraction of these, not only in immunocompromised patients but also in healthy people, may cause the respirators pathologies. The aim of this paper was to estimate the microbial contamination, in 20 hospitals located in three regions of the South Italy, for a total 81 operating rooms. The results show that 17 of the 20 operating units and 45 out of 81 operating rooms examined are contaminated. Periodic inspections should be carried out in order to control and lower the biological risk for both the patients and the health care workers. PMID- 18409938 TI - [Occupational exposure evaluation in some endoscopic services of Campania region sanitary structures]. AB - Peroxyacetic acid is an organic peroxide characterized by an elevated oxidative power, a remarkable biocide effectiveness (higher than the glutaraldehyde) and it can result cold germicidal at concentrations down to the 0.2%, too. In these last years, its use has been introduced in the sanitary sector in glutaraldehyde substitution (GTA), for the endoscopic material disinfection. The goal of this study has been to determine the aerodisperse peroxyacetic acid (APA) concentrations released by APA daily employment in endoscopic surgery operating rooms and sanitary endoscopy sections, related to some Campania region structures. A thirty samplings collection has been performed in "fixed posting", 15 measures in proximity of the "basins" and other 15 samplings in the closeness of the "automatic endoscope washer-disinfectors", to appraise the environmental pollution level during the whole working turn. Besides, 18 personal dosimetries have been effected in the APA solutions activation and decanting phases (load and unloading) employees: six "automatic endoscope washer-disinfectors" evaluations and twelve "basins" measures, for all through the operation (around 15-20 minutes). Air monitoring was performed by using vials containing alkaline silica gels with methyl p-tolylsulfoxide (MTSO) (1 l/min) and subsequently analysis by high performance liquid chromatography techniques (HPLC) equipped with a UV detector. The environmental investigations have underlined that APA average values result inferior to the specific TLV-TWA (around 1/10) and it reaches the same conclusion in comparison with TLV-STEL (around 1/4). Besides, the APA exposure founded in all the examined structures results smaller in comparison to the previously glutaraldehyde evaluation. PMID- 18409937 TI - [Prevalence of hepatitis virus A, B, and C markers according to the geographic origin of medical students]. AB - Antibodies against viral hepatitis A, B and C were evaluated in 221 students of graduate courses of the Medical School of Padua University born in countries different from Italy. Data were compared with those measured in 362 students born in Veneto Region and 87 students born in Centre-Southern Italy. The results showed a high, significant prevalence of positive antibodies against hepatitis A in students from Africa (94.7%), Asia (60.9%), Centre-Southern America (60.9%) and East Europe (52.7%); in Italy, the prevalence was significant in student from Centre-Southern Italy (19.5%). A high prevalence of hepatitis B antibodies was observed in 33.3% of Africa students (two subjects HBsAg positive), in 22.6% of students from East Europe (five subjects HBsAg positive) and in 12.5% of Asian and Centre-Southern American students (one HBeAg positive). Finally, infection with hepatitis C is sporadic and without a significant geographic distribution (three subjects only). PMID- 18409939 TI - [Tetanus vaccination and antibody tritation: compulsory vaccination at work and the role of the occupational medicine in the induction of protective effects in the population in general]. AB - In Italy in the last ten years tetanus incidence has been considerably decreased. It is a result first of the application of national laws that make compulsory the tetanus vaccine for some workers' group and for the population in general, and on the other hand for the working class as a consequence of D.Lgs 626/94. It was carried out by the U.O.O.M.L. of Garbagnate an activity of sanitary surveillance towards the working class with a tetanus biological risk (i.g. builder, metalworker, cleaning staff policemen, health visitors, cooks, etc.). During this activity it was considered the possibility of a tetanus vaccine cycle for those people that couldn't provide any certification about previous tetanus vaccine. The justification for this propose is to reduce the risk of side effects due to iperimmunization in case of close vaccinations and the safety and low cost of the tritation test of antibody anti-tetanus toxin method. Therefore it was decided to determine the personal condition of tetanus immunization for all those people without a valid certification. It resulted that 38% of people subject to sanitary surveillance were not protected and 55 years people resulted to be the group with highest risk. This result was also confirmed by the Ministry of Health that considers older population the highest risk group. It becomes clear that the vaccination for working purposes protects the population in general too. In conclusion we firmly think the procedure we followed completely accomplish the general principle of healthcare for infectious risk on the application of the tit. VIII of D.Lgs. 626/94. PMID- 18409940 TI - [The d.p.r. 37/97 and the thermal comfort of the surgeons according to the new version of UNI EN ISO 7730]. AB - The Italian law, through a specific government decree (d.p.r. 37/97), recommends the minimal technological, structural and organising requirements that surgery rooms of public hospitals must assure. Nevertheless, thermal comfort is usually evaluated by the indices PMV and PPD (Fanger's indices). Microclimatic data, acquired inside 80 surgery rooms over 20 public hospitals located in South of Italy, have been reviewed taking into account the new version of the international standard UNI EN ISO 7730. This version enables to classify the workers in three categories, A, B and C each one related to a different value of the acceptable number of dissatisfied people PPD. Considering the particular task which surgeons and nurses are involved in, a percentage of dissatisfied < 6% appears more realistic and conservative respect to the 10% prescribed in the old version of the standard. So the calculations show that, in this case, less than 17% of the staff would considered the thermal conditions as comfortable. Furthermore, the range of the microclimatic parameters, as established by the low, is not sufficient to assure thermal comfort using the PMV or PPD. PMID- 18409941 TI - [Prevalence of virus hepatitis B markers among medical students]. AB - Healthcare workers are a category of subjects at risk of infections higher than general population, though the introduction of vaccination against hepatitis B generally reduced this risk. In the present research, 2137 students of graduate course for health professions were subdivide in two age classes: 25 years old or younger and older than 25 years old according to the vaccination strategy defined by the law 165/91. A high number of students (82.3%) appeared immune against hepatitis B virus, whereas 16.7% were negative. About 30% of negative subjects declared vaccination. Females and subjects 25 years old or younger showed a significantly higher prevalence of anti-HBsAg antibodies (p < 0.001) than males and older subjects respectively. The obtained results show the need to evaluate the immunity status of the healthcare workers in training with the aim to reduce the number of susceptible subjects. PMID- 18409942 TI - [Tetanus risk prophylaxis: experience at work]. AB - As widely known Italian legislation makes the injection against tetanus compulsory for some units of workers. This isn't often easily organized and accepted. The aim of our work is to bring forward our experience with a group of workers servicing in the airport where tests have been carried out to establish the correct antibody cover through periodic controls. Only 46 workers, out of the 294 tested, have shown immunity deficiency. The vaccination programme has therefore been focused only on this group, realising a considerable organization and economic saving and a better acceptance from the workers side. Programmes have been drawn up for a future vaccinal programme taking into consideration the antibody cover results. PMID- 18409943 TI - [Well-being and safety awareness in the context of hospital units related to work place quality]. AB - Several studies founded a positive relationship between environmental work stresses, perceived safety of work places and number of injuries. This study tested the relationship between cognitive dimensions of environmental quality, dimensions of affective quality, and perceived safety in the context of hospital units. A total of 204 hospital workers filled in a questionnaire measuring Perceived Hospital Environment Quality Indicators (PHEQIs: Fornara et al., 2006), Affective Qualities of Places (Perugini et al., 20021, and the Work Safety (WSS: Hayes et al., 1998). Results show a positive relationship between subjective indicators (both cognitive and affective) of environmental quality and specific dimensions of perceived safety in hospital units. PMID- 18409944 TI - [Cognitive performances in workers with chronic virus hepatitis (HBV-HCV)]. AB - Chronic condition in subjects with chronic viral hepatitis determines issues neuropsychic. The sample of 21 workers suffering from chronic viral hepatitis in drug treatment has been studied with a battery of standardized tests to assess the cognitive performance, the neurobehavioral effects and psychological disorders that interfere with quality of life, comparing the results of subjects with HBV with those of subjects suffering from HCV. The results showed that both subjects with chronic HBV and HCV have relational-work restrictions that determine long periods of absence from the workplace, with the depression, anxiety, irritability and dysphoria. It is that in patients with chronic HCV physical functioning is significantly impaired with clinical manifestations of the disease that lead to major depression and deficit cognitive function. PMID- 18409945 TI - [Dosage of antitetanic antitotoxoid antibodies in the context of antitetanic procedures: case study including a group of Italian railways workers with obligatory antitetanic vaccination]. AB - The authors relate an half-year of antitetanic vaccination activity of the Health Office of Italian Railways in Milano. The operating protocol provided a preliminary dosage of tetanic toxoid in lack of information about previous vaccination. The results have underlined a good percentage of subjects who don't require further vaccination. PMID- 18409946 TI - [Biologic risk due to accident in academic personnel]. AB - Needlestick injuries since 2004 to 2006 were evaluated in University healthcare workers that reported an accident by point, sharp or mucosal contamination. During this period, 497 accidents with instruments contamined with biological fluids were reported. The injuries were most frequent between 9 a.m. and 1 p.m. (233 accidents). There is no difference during the week (excluding Saturday and Sunday), whereas February, May, June, and July were the months at risk. The most of accidents were during the first four hours of the job. They were identified 423 known sources and compliance with follow-up was evaluated. Only 26.3% of subjects injured with known hepatitis B source, 32.3% with known HIV source, and 40% with known HCV source completed follow-up. Fortunately, no seroconversion was observed. The lack of compliance with the follow-up, also if the source is known, needs to stimulate healthcare workers to subject to the protocols and to follow the standard procedure to prevent the needlestick injuries. PMID- 18409947 TI - [Virus C hepatitis: successful therapy in a health care worker]. AB - Hepatitis C viral is a problem of population health. The World Health Organization considers Hepatitis C an epidemic, a "silent" epidemic because a patient living with Hepatitis C can be infected for decades before being discovered. Recent data show an estimated number of 170 million patients infected with hepatitis C virus in the world. Number of new infections per year has declined from an average of 240,000 in the 1980s to about 26,000 in 2004. The incidence of HCV infected patients is estimated to 500-600 new cases in a year in Italy. Chronic infection is present in 55%-85% of infected persons. Approximately one third of the patients develop cirrhosis over a number of years, which can lead to liver failure and other serious complications. There is no vaccine and no completely effective treatment. Recent data show PEG-IFN-RBV combination therapy is most effective. We describe one HCV infected individual case report with HCV genotype 1b who received combination therapy for 4 weeks. Levels of HCV RNA became undetectable after an mouth of treatment. PMID- 18409948 TI - [Influenza vaccination in health care workers of a university hospital in Milan]. AB - The study analyses the percentage of influenza vaccination in health care workers of a big university hospital in Milan during the season 2006/2007. The percentage of vaccinated subjects was 19.8%. The frequency of vaccination has also been assessed according to sex, age, job and school title. Observed data indicates that the number of vaccinated sanitary workers is far below the levels reached in other European countries and in the USA. Thus, in order to reach a higher adhesion to vaccination programs, sensitisation campaigns are necessary. PMID- 18409949 TI - [Management of latex allergy in the Pisa University Hospital: analysis of data deriving from the Registry filing both allergic patients and employees]. AB - Recently, Tuscany regional government has promulgated (Regional law n. 464/07) guidelines for setting up suitable sanitary rooms for patients allergic to latex. These guidelines enforced the managers of the Tuscany Local Health Units to promote educational activities and a correct methodology for the safe management of both health workers and patients who need latex-free diagnostical and/or therapeutical procedures. An other Regional law (n.345/05) had previously enforced all the Tuscany Local Health Units managers to implement and update a Registry of every latex-allergic patients ever hospitalized in their own local facilities. Since 2002 a multi-disciplinary task force of health operators, lead by the Preventive Medicine Section, has been elaborating and implementing a plan for the safety of latex-allergic patients of the Pisa University Hospital, along with a Registry where each of these patients were recorded. Moreover, since 2005 under the responsabilities of the staff of the Preventive Medicine Section, another Registry has been implemented and updated. In this Registry all the Hospital employees who have been diagnosed a latex allergy, through the medical exams performed within the occupational medical surveillance activity, have been filed. In this study we report the results of this preventive activity both aimed at patients and heatlh operators safety. PMID- 18409950 TI - [QuantiFERON-TB Gold test in homeless shelter staff: preliminary results]. AB - Tuberculosis (TB) continues to be a major cause of disability and death and it has become a real problem in industrialized countries. The spread of HIV, the increasing immigration rate of people from countries with endemic TB and the growth of drug-resistant Mycobacterium tuberculosis strains extend its impact. Since the spread of the infection occurs early, health care workers are particularly exposed to the risk of contracting and/or transmitting the mycobacterium. In health care settings, policies and procedures for TB control should be developed, including health surveillance. Until recently the tuberculin skin test was the only available method for diagnosing tuberculosis, however it suffers several methodological weaknesses: high rate of false positive results in vaccinated populations, the subjectivity of the evaluation and the booster effect. Recently, the introduction of new in vitro serological tests, as the Quantiferon TB-Gold in tube (QFT-TB), may overcome these problems. The QFT-TB is based on the quantification of interferon-gamma released from sensitized lymphocytes in whole blood incubated overnight with PPD from M. tuberculosis and control antigens. The present study was performed on 27 nuns (homeless shelter staff) who were at risk for contracting tuberculosis. The prevalence of positive tests was 15 out 27 (55.5%). PMID- 18409952 TI - [Methodology assessment of the first-aid in a company]. AB - The Ministerial Decree 388/03 establishes the first aid duties and organisation. It classifies the health firms into three groups, according to the types of activities carried out, to the number of the employees involved, and to the risks linked to it. The membership of a firm to a group reflects upon its standard equipment of facilities, the training required for the employees and the duties of communication to carry out to the Health Service. During the activity of vigilance we have verified that a lot of firms, belonging to our territory, have not applied correctly the body of rules regarding the first aid. For this reason a project aiming at providing information about the application of this decree has been started. PMID- 18409951 TI - [Antitetanic vaccine in a population workers of Valle d'Aosta: fire volunteers department experience]. PMID- 18409953 TI - [Assessment of radiofrequency electromagnetic fields levels in the Taranto area]. AB - Radiofrequencies are extremely common in telecommunication sectors, in industry, in medicine and domestic applications. We carried out an observed study focusing on the punctual monitoring of the electromagnetic fields generated by mobile phones handsets or by SRB radiobase stations (transmitters), installed in Taranto and throughout the borough council. The measurement campaigns have been done by ARPA Puglia, Provincial Department of Taranto from November 2005 until March 2007, on 108 transmitting stations, of which 72 are of single installation and 36 of multiple installations, with UMTS, GSM. DCS, DVB-H kind of communication. The monitoring has been done with Wandel & Goltermann EMR-300 devices; the registered frequency is within the interval between 100 KHz and 3GHz and measures have been obtained with precise time interval of 6 minutes. In most of the monitored areas the results respected limit values; only in 2 cases on 108 the attention value has been exceeded, both within borough council area. In the town of Taranto values over 6 V/m fixed by the actual rule haven't been registered in none of the 51 monitored stations. We can, therefore, define within the limits the expositions emitted by the radiobase stations and in the buildings nearby the antennas. PMID- 18409954 TI - [Measurements of professional exposure to ELF fields in some production areas within the territory of Regione Campania and comparison with the action values according to 2004/40/CE Directive]. AB - The 2004/40/CE Directive (which should be acknowledged within April 30, 2008), introduces the Action values for the prevention of occupational risk related to Extremely Low Frequency-Magnetic Fields (ELF-MF) exposure. In the Regione Campania, the following production areas have been investigated: textiles, industrial graphics, wood, manufacturing and ceramics. We have specifically monitored exposure to ELF magnetic field values for 400 workers belonging to 80 different areas chosen among those for which an exposure could be supposed. Results have been compared to the action values indicated in the 2004/40/CE directive. Used instrumentation includes a Wandel & Goltermann EFA 300 portable field analyser with B-Field Sensor isotropic spectrum analyzer for electric and magnetic field with 5 Hz to 32 kHz bandwidth. Technical rules follow CEI 211-6 Jan-2001. All measurements have been performed in continuous within two working shifts, setting the instrument to make acquisitions every five minutes. Data have been then processed to obtain RMS values of the magnetic field per each work shift. Values found have not shown any exceeding of the action values: more specifically, 90% of values was below it (500 microT) and 70% of the monitored jobs shows an occupational exposure below 0.4 microT, which is the possible threshold proposed for the man chronic effects of ELF magnetic fields based on data coming from some epidemiological studies. PMID- 18409955 TI - [Work related disease in radiation exposed? Case report]. AB - In the present work we analysed the possible recognition of professional disease in a subject exposed to ionizing radiation. Haematic parameters were evaluated during the working activity and after the risk's avoidance. Normal haematic values after the pathogenic noxa's avoidance exclude a chronic damage, which in an indispensable datum for professional disease recognition. PMID- 18409956 TI - [Radon exposition preliminary research in a Sicilian territory]. AB - Radon toxicity on human body is well known from along (in 1988 radon has been classified as first type carcinogen, after only to tobacco's smoke, as cause of lung's cancer). Based on known scientific data, preliminary study has been conducted by the AA. It concerns radon exposition on inhabitants living in a Sicilian territory featured by previous seismic events: the territory and the town of Montevago. The project has been sponsored by ARPA Palermo. The territory of Montevago has been divided in several areas in order to assign detectors homogeneously, to begin the environmental sampling. In the period between May and October 2006, instruments has been calibrated and standardization of the procedure has been completed, in collaboration with Centro Studi Nucleare Enrico Fermi del Dipartimento di Ingegneria Nucleare del Politecnico di Milano. The values obtained result in European range. PMID- 18409957 TI - [Evaluation of the occupational risk related to exposure to electromagnetic fields according to the EC Directive 2004/40 EC: exposure during pregnancy]. AB - The EU Directive 2004/40/EC on the minimum health and safety requirements regarding the exposure of workers to the risks arising from physical agents (electromagnetic fields) will introduce the need of an evaluation of the risk related to EMF occupational exposure in pregnancy. Nevertheless, data from research in this field are scarcely conclusive to date. Furthermore knowledge on this risk seems insufficient among OH physicians in Italy. Accordingly, there is an urgent need for further research, and for a diffusion of knowledge among OH physicians on possible risk to pregnancy due to occupational exposure to EMF. PMID- 18409958 TI - [Electromagnetic interference of electronic devices in workers with AICD]. AB - The Implantable Cardioverter Defibrillators and the Pacemakers are devices that produce electric signals and consequently they are touchy to the electromagnetic interferences. This touchiness may cause a temporary or permanent device's malfunction both for short, but of high intensity, and prolonged exposures. Considered the wide diffusion in the job's environments of the technologies based on the electromagnetic waves, the aim of the study is identify the presence of electronic equipment's electromagnetic interferences in AICD bearing workers. The last aim is give to the AICD bearing workers more detailed information about the prevention of inappropriate AICD shocks. PMID- 18409959 TI - [Radiation protection issues in brachytherapic treatment of prostatic cancer]. AB - Brachytherapy is an effective radiotherapeutic treatment for localized prostatic cancer. The permanent brachytherapy is a particular kind of radiotherapy which, US guided, uses permanently implanted seeds containing radioactive sources (Pd103 or I 125). The procedure is minimally invasive and allows to obtain high percentage of success which is comparable to surgery. The possibility to confine permanent radioactive implants in a well delimitated area doesn't exclude the exposition of both the medical staff and family's members of implanted patients. The radiation exposure involves the medical physicists, the radiotherapists, the Medical Radiology Technician, the anaesthetists, the surgeons, the professional nurses but also, after the brachytherapy treatment, public and family members, comforters and cares. It's necessary to consider radiation safety aspects of brachytherapy in order to reduce the risks of exposition. At the end, several studies showed that cremation of bodies could be a possible radiation source that remains in the patient's ashes, potentially inhaled by crematorium staff or members of the public. PMID- 18409960 TI - [Health promotion and computer science in radiation protection]. AB - An automatic system of clinical-diagnostic information has been applied to workers exposed to ionising radiation at the University of Naples Federico II with reference to the last 5 years. For every person exposed a computerized case sheet was elaborated recording clinical, biological, dosimetric and other preventive data. In the localized risk, capillaroscopic monitoring was used. This research has highlighted the role of medical surveillance in developing health promotion criteria and the planning of the interventions with the complete control of all data in real time. PMID- 18409961 TI - [Operational aspects of risk perception in the electromagnetic fields exposition]. AB - The increase of electromagnetic fields exposition is being associated with the increase of risk perception in the people exposed due to the uncertainty of the biological and sanitary effects. Research is being carried out on the symptomatology shown by the 45 people living near power-lines in the Benevento area and consequently exposed to electromagnetic fields. The measure of the magnetic and electric field was in the normal range while the people showed most symptoms in the subjective and behavioural sphere. The research findings suggest that risk assessement should take into account the subjectivity of the people exposed as shown in the special questionnaires with the aim of reducing the subjective and behavioural symptomatology for developing a new environmental medicine. PMID- 18409962 TI - [Radioprotection optimization measures in hospital workers]. AB - In this contribution, a revision of ionizing radiations exposed workers' dosimetric data in four hospital structures is showed, and, consequently, recommended optimization measures are identified. PMID- 18409963 TI - [Analysis of the natural radioactivity due to the radon gas in the underground of Rome]. AB - BACKGROUND: Radon is a decay product of 238Uranium which is classified by WHO/IARC as group 1 carcinogen, given its causal relationship with lung neoplasia. An annual concentration of this gas higher than 500 Bq/m3 in workplace is considered potentially dangerous by the italian legislation. No data are currently available on radon level in underground tunnels, which are a potentially important source of exposure both for workers and travellers. MATERIAL AND METHODS: Measurements have been performed in a station and within the trains. Two months integrated measures, and 5 days continuous (hourly) assessments have been performed. Integrated measurements have been performed by means of 12 passive dosimeters, containing a detector made of CR39 (polymeric type), whereas active scintillation-type dosimeters have been employed for continuous assessments. RESULTS: Two months integrated measures: radon level in the station was 665 +/- 71 Bq/m3, whereas values within the trains ranged between 96 and 117 Bq/m3. Continuous measurements: Mean radon level during work activities was 783 +/- 536 Bq/m3 and thereafter it rose to 850 +/- 131 Bq/m3 CONCLUSIONS: Radon nelle levels in underground tunnels may exceed attention levels, whereas within trains they remain well below these levels. Further studies focused to assess the risk of underground employers are needed. PMID- 18409964 TI - [The activity of the Service for Prevention and Safety in Workplaces of the Health Local Agency of Rome C towards the "competent physicians" in the enterprises of the territory]. AB - In the past seven years the Service for Prevention and Safety in Workplaces of the Health Local Agency of Rome C set out some activities towards the occupational physicians (so-called "competent physicians") of three different kinds: control and surveillance, institutional activities on demand and activities for quality improvement. In the first ambit we conducted some investigations on the procedures of the workers' medical surveillance and the role of competent physicians in the risk evaluation inside three different intervention projects (monitoring the enforcement of the D.Lgs. 626/94; intervention on the chemical risk, intervention in the car repair shops). In the second ambit we analysed 92 notifications of occupational diseases and we dealt with 27 appeals against the judgement of the fitness to work. In the third ambit we made 19 meetings on various subjects: correct procedures in medical surveillance, quality in diagnostic examinations, the enforcement of new regulations. We report the results of these interventions which let us identify the most common criticalities in the activity of the competent physicians, but also to face them with a new approach based more on peer review instead of control and surveillance. PMID- 18409965 TI - [Vigilance on health surveillance in wood sector]. AB - OBJECTIVES: A program to promote occupational health among wood dust workers in the district of Civitavecchia. METHODS: In recent years, occupational health physicians charged of medical surveillance of wood workers (Competent Physicians, CPs) had been invited to perform a peer-review of their methods and activities. In the present phase, CPs have been invited to show the result of their medical surveillance. RESULTS: One hundred forty seven wood workers were submitted to rhinoscopic examination. The prevalence of woodwork-related rhinitis and other pathologic signs, including nasal adenocarcinoma (one case), was 32.7%. The prevalence of rhinitis in woodworkers increased with years of working as a woodworker. CONCLUSION: Wood dust and chemical exposures in wood workers represent a serious risk of disease for the nasal cavity and paranasal sinuses. PMID- 18409966 TI - [Smoking: results of an interventation for health education of young workers attending to the SPISAL Service for pre-employment medical examination]. AB - Young people who start smoking are constantly increasing and the age of initiation has decreased. This work presents the results of a specific health education programme on smoking that was carried out for a group of 804 young apprentices who came to the Occupational Health Units in Belluno for a check on work fitness. The objective is to induce young non-smoking apprentices to continue as such and smokers to give up smoking. After two years, a telephone follow-up was carried out to see if the smoking habit has changed. The average age of the group was 19 years and the percentage of smokers was 34%; after two years the percentage of smokers was 29.5%. Then, the percentage of smokers has fallen and the percentage of ex smokers has increased, because smokers became non smokers during the period of observation. This results show the effectiveness of health programme on smoking directed to young apprentices already demonstrated by a previous study and confirm the withdraw rates obtained by counselling methods. Moreover, this study stressed the role of Occupational Health Units in prevention programmes addressed to the young apprentices during work fitness check. PMID- 18409967 TI - [Interaction between occupational health service and National Health Service. Appeals against duty ability judgements]. AB - The main purpose of the medical surveillance, in conformity with the Italian D.L. 626/94, is the formulation of ability judgements. The above-mentioned D.L. 626/94 gives the workers the possibility to file a petition versus the judgement itself addressing it to the ASL concerned. The aim of the present work is to report experiences made with the occupational health service as for versus appeals. About 2000 examinations a year are carried out at our occupational health service and the concerned judgements are pronounced. 29 versus appeal have been forwarded between 2002 and half 2007. The ASL decision has been favourable to the "competent" doctor judgement in 21 cases, and adverse in 8. Most appeals concerned osteoarticular problems. Finally some remarks are made about the necessity of improving the cooperation between "competent" physicians and supervision national health services. PMID- 18409968 TI - [Occupational safety and health in hospitals of Regione Lombardia. Are there differences among public, private nonprofit and private profit hospitals?]. AB - In 2006 an extensive survey took place in all hospitals of Lombardia, to assess, through inspections, the safety conditions and the efficiency of the organisations invested to assure that. During these inspections were evaluated 6 safety sections and hospital organization and efficiency, by means of a questionnaire. The aim was to assess the internal consistency of questionnaire sections, the presence of differences of such scores among public, private non profit and profit hospitals and the correlations among safety scores and among these latter and efficiency indexes. Significant differences for size factors and efficiency indexes have been pointed out among different types of hospitals. Instead, safety scores showed good internal consistency and have been resulted statistically significant correlated in most cases. For all types of hospital have been observed high values for safety scores, with exception of occupational health service, better organized in public hospitals, and of safety conditions of worker from external agencies, which have been displayed low. This last consideration stick out the need to pay more attention to safety conditions of these workers. PMID- 18409969 TI - [Occupational counselling as a potential tool for evaluating safety management at health facilities]. PMID- 18409970 TI - ["Barber's hair sinus": new clinical observation]. AB - The AA describe an unusual case of pilonidal sinus in a barber. The clinical analysis showed the presence, in all digital spaces, of erythemateous areas irregularly infiltrated by small fistular tracts and a keratinization area. In the first interdigital space of the left hand, the discharge of a bristle from the apex of a papule was also visibly evident. The histological examination showed the presence of a foreign body granuloma, with giant cells, lymphocyte and plasma cells, centered on a bristle fragment immersed in a collagenous matrix with abundant keratine, skin thickening and fibrous dermis. Using a probe, it was possible to follow the fistular path and obtain the discharge of several hair fragments, conglutinated by corneous scales, but it was not possible to achieve clinical cure of the lesion. The solution to the problem therefore required surgical therapy. PMID- 18409971 TI - [Dysphonia and cacosmia in a worker in sterilized rooms]. AB - A 39 year male pharmaceutical worker employed in a clean-room developed in 2003 acute dysphonia after environmental disinfection with glutaric aldehyde and isopropyl alcohol. Laryngoscopic examination showed glottis edema; the syndrome healed after a cycle of cortisone. In subsequent years, withdrawal from exposure to irritating chemicals was observed. The worker, however, complained for recurrent episodes of dysphonia, in the absence of abnormalities of the larynx, and gradually developed intolerance for perfume, solvents, and other smelling substances. He came to our observation in 2007, showing strong conviction that occupational exposure had a causative effect in his complaints. He was working in an office open to public, and fragrance exposure appeared to be the main cause of dysphonic episodes. We rejected the hypothesis of association between complaint and job exposure, and advised him to work in a well defined working situation, such as in a clean room, where chemical contamination may be effectively controlled over time. PMID- 18409972 TI - [Adenocarcinoma of the nose after occupational exposure to wood dust: description of a case]. AB - The occupational exposure to wood dust, classified by IARC as carcinogenic certain may determine the onset of epithelial tumors of the mass graves and nasal sinuses. The symptoms of these malignancies are aspecific (nasal obstruction, rhinorrhea), the diagnosis is often delayed. CLINICAL CASE: Worker of 59 years occupied, from 1960 to 2005, in carpentry (construction of coffins), who died in 2006 for a nasal adenocarcinoma of the mass graves extended to the cranial base, initially considered intracranial cancer given the wide cranial fossae prior to the histological diagnosis. The case is emblematic to late diagnosis and long survival (the worker has survived for more than three years after TAC diagnosis of intracranial mass occurred in 2003). CONCLUSION: Extending the cranial base of a cancer of the nose and sinuses occurred in a worker exposed to wood dust is a serious critical in the system of health surveillance in the company. This rare neoplasm is a "sentinel event" that highlights a problem of underestimation of this occupational tumor and consequent failure of occupational disease complaint is therefore required a careful accounting and reporting of cases. PMID- 18409973 TI - [Occupational stress and job insecurity in men working in a university]. AB - 77 men working in a university were investigated. Trait and state anxiety were determined by STAI I and STAI II; job strain (job demand/decision latitude), social support and job insecurity were analysed by a 46 item Karasek's questionnaire and subjective symptoms by a 12 item test. The employees of a library (mean age 49 years), in contact with students, showed significantly higher values of job strain, STAI I, STAI II and subjective symptoms than a control group of employees with similar age. Young employees and sanitary staff with temporary employment showed higher level of job insecurity than control subjects with stable position. Blood cytotoxic activity (reported in another study) was significantly lower in the old employees with job strain or in the young employees with job insecurity (but not in the sanitary staff) than in the controls; this demonstrates that not only occupational stress but also job insecurity may play an important role in affecting the health status. PMID- 18409974 TI - [Job strain and blood pressure levels in health care workers]. AB - OBJECTIVE: The aim of this study was to investigate a possible association between job strain and blood pressure levels in a sample of X-ray technologists. MATERIALS AND METHODS: A structured questionnaire S was administered to the participants, investigating individual factors and job characteristics. Job strain status was measured by means of questions derived from the Italian version of the Karasek Job Content Questionnaire. Systolic and diastolic blood pressure were then assessed during medical examination. RESULTS: No significant association was found between job strain and blood pressure levels. In addition, no association was found between high job demans, low decision latitude and blood pressure levels. CONCLUSIONS: In this study job strain was not associated with high blood pressure levels in X-ray technologists. PMID- 18409975 TI - [Ticks bite in foresters]. AB - The objective of the study is evalutation of the risk for ticks strings on foresters. The sample constituted by 325 foresters belong to Messina province as been submitted to medical examination venous tests. Whole sample had to answer to a questionnaire to consider. The prevalence of systemic and skin reactions and we have dose Immunoglobulines versus Brucella Melitensis, Rickettsie Conorii e Borrelia Burgdorferi. The results showed that the 19% has declared past stings of tick, and 4.9% reported symptoms probably deriving to a past infections determined by inquired microorganisms. The serum tests showed that 70% was positive for all microorganisms, instead only 31%. Was never infected by inquired microorganisms. In conclusion our study shows that zoonos is risk linked to stings of tick is relatively high in foresters. PMID- 18409976 TI - [Evaluation of the occurrence of a rare work related disease in a cohort of workers employed in the processing of paper]. AB - The aim of our study is evaluating occupational exposure in a factory named "Poligrafico Zecca dello Stato" in Foggia. The workers had a occupational disease "chlorine related parodontal disease" indemnified by INAIL (Italian Occupational Assurance). The study demonstrated the workers have been indemnified worked as engineer, drying and electrolysis process employed. The study showed that the workers indemnified without legal action were different in respect of workers that take legal action, because the last showed, paradoxically, a chlorine occupational exposure. The explanation of this paradox might be the production cycle non acknowledge by operators. PMID- 18409977 TI - [Work outcome evaluation of cardiac surgery patients after rehabilitation training]. AB - The aim of this study was to investigate the incidence of job resumption and the influence of clinical, psychological and functional factors in patients who underwent cardiac surgery. 859 patients were evaluated (mean age 66 +/- 9, 72% M, 28% F, 75% retired, 7% housewives and 18% still working) following admission in a Cardiac Rehabilitation Center; within 5 days of cardiac surgery. Six months after surgery, 80% of the patients resumed their previous occupation. The variables which positively influenced the latter were: a) left ventricular ejection fraction > 35% (p = 0.002), average-high education (p = 0.002), younger patients (p = 0.032), absence of complications during the 6 months post-surgery (p = 0.001), having attended frequent educational programs during the rehabilitation period (p = 0.022), a precise knowledge of the illness (p = 0.001), lack of depression (p =0.0001) and anxiety (p = 0.0001). Our data confirm, in accordance with the recent guidelines, the importance of a rehabilitative intervention which comprises physical, educational and psychological support. PMID- 18409978 TI - [Occupational brucellosis in the veterinary service of the Local Health Service in the Abruzzo Region (Italy)]. AB - During the health surveillance of the employees in the veterinary service of a Local Sanitary Service, we came across a medical case of occupational brucellosis in a veterinarian, identified by seriological data (Wright' seriological diagnosis + brucellosis specific test with identification of IgM and IgG), in a lack of significant anamnesis, anamnestic negativity of dangerous behaviours. Two medical cases of the sole seriological positivity have been identified in a veterinarian and in a health vigililant. In our Local Health Service the medical cases stated across the population have been 6 (in the period from 2000 to August 2007). The epidemiological flow of the animal brucellosis in the regional area, and in particular in our territory, shows one low incidence of positive stock farms, low incidence of oxes, sheep and goats infected. In Italy the medical cases of brucellosis denounced and approved from the National Institute for work related Injury Insurance have been 38 out of 41, in the period from 1996 to 2006. This zoonosis represents a problem of relief in the sector the breeding of the animals and the butchery of the meats in all countries; the occupational health physician, within the area of prevention, has to exercise a specific health surveillance of biological risk and take part in specific programs of information and formation, in particular on the use of individual protection devices and correct behavioural rules. PMID- 18409979 TI - [Chronic fatigue syndrome: from misunderstood illness to cause of job fitness management problem and of work total disability in a physiotherapist. Case report]. AB - The aim of this paper is to report the case of a Physiotherapist working in a big hospital, affected by Chronic Fatigue Syndrome (CFS). After the diagnosis, made in an High Specialized Center, the Occupational Health Physician, with the cooperation of the Nursing Managing Direction, the Chief of the Department of Rehabilitation and the Physiotherapists Coordinator, had to cope with the job fitness management. Afterwards the patient, in accordance with the Physician of a Trade Union Medical Office and the Occupational Health Physician, tried to obtain the disability pension, that at the end was given by the Medical Commission of the ASL. PMID- 18409980 TI - [Cardiorespiratory fitness and arterial stiffness in sedentary and not sedentary hypertensive workers]. AB - In hypertensive subjects, arterial stiffness and arterial wave early reflection are thought to be the major factor limiting cardiac reserve and exercise capacity. Aortic augmentation index (AIx) is a measure of arterial wave reflection and stiffness, and has been associated with decreased cardiorespiratory fitness. We investigated the role of physical activity at work and its effect on such association. 25 hypertensive middle-aged workers, without history of diabetes, cardiovascular disease, renal failure and inflammatory diseases, were studied. Our study confirms that AIx provides information for the prediction of VO2 peak, being also gained with a non-invasive and practical test. Inside the two groups of sedentary and non sedentary workers, the relation between AIx and VO2 peak is still significant; whereas the non sedentary group showed a worse cardiorespiratory fitness without significantly differences in arterial stiffness. PMID- 18409981 TI - [Good practices and techniques for prevention of accidents at work and occupational diseases. New database of Inail]. AB - The project "The publication of good practices and good techniques for prevention" is one the priorities of nail. This computerized system for the collection of good practices and standards of Good Technology is aimed to health and safety of workers. The basic objective of the database is to provide a valuable tool, usable, dynamic and implemented, in order to facilitate and direct the access to BP and BT it by people responsible for SSL. At the same time constitutes a tool strategically important for enterprises (especially SMEs) in terms of technological innovation and competitiveness, related to the prevention, safety and health of workers. The realization of this project has involved many of the professionals (chemists, engineers, doctors, biologists, geologists, etc.), and everyone gives his intake of qualified professional competence. PMID- 18409982 TI - [Air quality and human health: PM10 and respiratory diseases]. AB - In this paper a comparison between the air quality data collected in urban and rural areas is performed taking in account the PM10 levels allowed in the European Union and in the world and is related to the increase of respiratory diseases. The parameters considered are essentially due to the anthropogenic primary pollution: particulate matter PM10, benzene and toluene, CO and Polycyclic Aromatic Hydrocarbons. PMID- 18409983 TI - [Respiratory risk among students in an industrialized area of Sardinia: role of smoking and air pollution]. AB - Among 273 students (age 11-16 years) living in two small towns located respectively 2 (Portoscuso) and 15 kilometres (S. Antioco) far from the industrial area of Portovesme, the respiratory risk associated to cigarette smoking and environmental pollution was evaluated by the ISAAC questionnaire and standardized spirometry. The prevalence of parent's smoking was high in both groups (66.3% and 59.4%). Smoking during pregnancy was reported by 15% of mothers, without significant differences between the two schools. The prevalence of smokers among students was similar comparing the two groups (7% vs 6%). The prevalence of asthma in the whole population was significantly associated to skin positivity to common allergens, to the familiarity for asthma and to the environmental tobacco smoking, particularly if maternal. Spirometric values were significantly lower among students living in Portoscuso than among those of the S. Antioco school, with an evident interaction between the residential factor and the tobacco smoking, active and/or passive, in reducing the forced end- expiratory flows. Our results support the significant role of tobacco smoking, active and passive, particularly if derived from maternal smoking during pregnancy, in increasing the prevalence of respiratory disorders and lowering lung function in children. Living in Portoscuso, because of higher environmental exposure to airborne pollutants of industrial origin, has been shown as a relevant factor further lowering the lung function among the studied sample. PMID- 18409984 TI - [Efficacy of preventive measures for reducing professional disorders due to pumice powder]. AB - The analysis of the incidence of pumice pneumoconiosis in the period 1970-2002 allows to evaluate the positive effects, particularly on the deadly forms of the disease, of the technical interventions aimed at reducing dusts. The main interventions in the field of the workmanship of pumice were made at the beginning of 70s, with the introduction of electrofilters in the working cycle and the change of the pumice desiccation system, from plate ovens to closed-cycle rotating ovens. This eliminated the need for manual shedding of pumice stones on oven plates. That duty had caused, between 70s and 80s, an high incidence of deadly forms of pneumoconiosis, which dramatically decreased with the reduction of workers exposed to the above risk. A comparison between the period 1970-75 (before preventive technical interventions) and the period 1976-2002 clearly shows an evident decrease both of deadly forms of pneumoconiosis and of acceptance of appeals for professional silicosis. PMID- 18409985 TI - [Respiratory function and smoking habit among shipyard and dock workers]. AB - The aim of the study is to compare the loss of lung function in a group of 196 male workers employed in shipbuilding/repairing and port activities ("cases") with a group of 109 healthcare workers ("controls"). "Cases" were grouped by radiological examination (chest X-ray/HRCT) depending on the presence or absence of pleural plaques. Smoking habit was assessed in all subjects. Lung function assessment included two measures of VC, FEV1 and TLC (the latter for "cases" only). The mean interval between the two examinations was five years; changes in spirometric results (value at second examination minus value at first) were expressed as ml/year. The results showed that there was no difference in loss of lung function in the three groups over the follow-up period, even when differences in smoking habit were taken into account. The results suggest that radiological findings of pleural plaques had no bearing on deterioration in lung function. Lung function testing is widely accepted as an integral part of medical surveillance of occupational lung diseases, but the high inter and intra-subjects variability of spirometric data underlines the difficulties and limitations of lung function follow-up studies. PMID- 18409986 TI - [The risk of inhalable wood dust: assessment of workers exposure wood working factories]. AB - The International Agency for Research on Cancer RC) has classified wood dust as carcinogenic to humans based on demiological and experimental evidence. Exposure to wood dust may use respiratory and dermal symptoms and diseases. The aim of this work was to estimate occupational exposure to inhalable wood dust adopting the formal procedure described by UNI EN 689/97. The exposure of 23 workers in three different working day was measured. In total, 69 personal air samplings were carried out at five wood working factories. Inhalable fraction of airborne dust was collected on 5 microm pore size, 25 mm diameter PVC filters utilizing the IOM samplers. The quantity of the wood dust was determined with gravimetric method. The results show that about 13% of the exposure values exceed the limit of 5 mg/m3 specified by the Italian Law Decree 66/2000 and about 48% of personal exposures are lower then the limit value. Prevention measures, technological solutions and personal protection equipment should be adopted in order to reduce worker's exposure. PMID- 18409987 TI - [Study on prevalence of upper and lower airways disorders in woodworkers, using data from medical surveillance reports and exposure registers]. AB - OBJECTIVES: To evaluate correlations between exposure to wood dust, upper airways symptoms and lung function. METHODS: We have analysed medical surveillance reports of 197 woodworkers with a median wood dust TWA exposure of 2.1 mg/m3. Every worker was examined by an otorhinolaryngologist and had a spirometric test. The results have been analysed with logistic regression to correlate prevalence of symptoms and spirometric data with occupational exposure to wood dust, length of service, regular use of respiratory protection and smoking habits. RESULTS: Epistaxis (prevalence: 10.1%) correlates with no smoking habits (OR = 6.4; p = 0.01); subacute or chronic rhinitis (prevalence: 41.6%) correlates with exposure to wood dust (O = 1.37; p = 0.01) and no use of respirstory protection (OR = 1.68; p = 0.09); subacute or chronic pharyngitis (prevalence: 17.2%) has a weak but significant correlation with length of service (OR = 1.03; p = 0.05); decrease in FEF25-75 (prevalence: 19.8%) correlates with no use of respiratory protection (OR = 2.56; p = 0.02) and exposure to wood dust (OR = 1.29; p = 0.09); pathologic decrease of VC (prevalence: 5.1%) correlates with exposure to wood dust (OR = 1.69; p = 0.05). CONCLUSIONS: this study seems to confirm that chronic irritation of upper and lower respiratory tract are caused by exposure to wood dust below the european 8 hours exposure legal limit of 5 mg/m3. PMID- 18409988 TI - [Burden of COPD on health profiles in different work activities]. AB - According to WHO, COPD will be the 5th cause of disability and the 3rd cause of mortality by 2020. Even cigarette smoking represents the main cause of COPD recent studies demonstrated positive association with occupational exposure. In Europe costs of COPD were estimated at about 38,8 billion Euros in 2000; its burden is also expected to increase. Aim of the study is to evaluate the burden of COPD on health profiles in a sample of workers of Campania region. Prevalences were calculated in a sample of 908 male workers, employed in industrial (479) and tertiary (429) sectors. Results were compared with population data of ISTAT database. Higher prevalence of COPD was found in industrial workers (23.4%) than general population (3.5%) and tertiary workers (2.1%). Analysing smoking habits, higher prevalence of smokers in the industrial sector (75.9%) than tertiary (66.4%) was found, but this difference cannot explain the big difference of COPD prevalence between the two groups and suggest a positive interaction between smoking and occupational exposure. In conclusion, the results analysis underlines the burden of COPD on workers' health status, particularly in some work activities. Specific health promotion programs are necessary in these activities. PMID- 18409989 TI - [Obstructive sleep apnea syndrome (OSAS) and work]. AB - Obstructive Sleep Apnea Syndrome (OSAS) is a very common disease in work age. Aim of study is to assess the impact of OSAS in a workers population. 138 workers (M 117, F 21), age 35-65 (mean 52.66 +/- 3.042) consecutively referred to Respiratory Hospital Monaldi and to Occupational Health Medicine Department of Second University of Naples performed an anthropometric evaluation of BMI, neck and an overnight polisomnography with Embletta X10 (Flaga Medical Devices; Reykjavik, Iceland). Workers' population was divided into three groups according to the impact of daytime sleepiness on work efficiency. Occupational Health Medicine needs to evaluate the high prevalence of obesity and metabolic syndrome (OSAS, diabetes, insulin-resistance) in work age population. PMID- 18409990 TI - [Acute effects of environmental pollution on the urban vigilants airways]. AB - AIM OF THE STUDY: The purpose of this study was to evaluate whether the acute exposure to air pollution, in a group of policemen of Padua, is correlated with increased inflammatory biomarkers (exhaled nitric oxide, feNO) and alterations of bronchiolar cells (assessed by CC16 Clara cell-specific protein). METHODS: We studied 44 healthy, non-smokers divided in exposed to traffic and controls (office workers). Before and after the Monday shift serum and urinary concentration of CC16, feNo and spirometry were measured in each subject. Data on air pollutants, PM2.5, PM10, SO2, NO2, CO, O3 were collected from official bulletin online (ARPAV). RESULTS: In exposed policemen serum CC16 decreased after shift (before 4.6 +/- 0.2 vs after 6.4 +/- 0.8 ng/ml, = 0.02), while feNO increased significantly (33.2 +/- 4.4 vs 29.7 +/- 3.9 ppb, p = 0.02). feNO cross shift changes were positively correlated with environmental SO2 levels (rho = 0.48; p = 0.01). CONCLUSIONS: Our results suggest that in healthy and nonsmokers subjects the exposure to air pollution is associated with subclinical airway inflammation and decrease of bronchiolar epithelium function. PMID- 18409991 TI - [A case of occupational lung disease (welder lung) in a mechanical worker]. AB - A case of welder's lung is reported in a male hard smoker who had worked as welder-carpenter for almost 30 years. In order to establish an etiologic diagnosis, given the informed consent from the worker, open lung biopsy was performed that allowed to obtain sufficiently large specimens of affected lung to make histological, immunohistochemical and mineralogical examination. In such a way the pathologic features were accurately defined and a multidisciplinary approach was applied to differential diagnosis. The worker is recommended to undergo a strict health surveillance because of the reported association of welder's lung with lung cancer, especially in smokers. PMID- 18409992 TI - [Role of the occupational health physician in the evaluation of biological risk and cancer in a hospital setting]. AB - OBJECTIVE: Elaborating and proposing a model of behaviour which is useful for any occupational doctor of a hospital in the management of the people exposed to biological risk, with the aim of preventing or early diagnosing neoplasias caused by cancerogenic infective agents. MATERIALS AND METHODS: This study was conducted analyzing the literature data regarding biohazard work accidents which happened in health environment and the scientific evidence of the causal relationship between infective agents and development of neoplasias. RESULTS: Data in literature show that the biohazard work accidents are very numerous and there is high percentage of sub-communication of them; many infective agents that the health workers can get in contact with, after biohazard biological accidents, are cancerogenic: HBV, HCV, HIV, HP belong to group 1 of IARC classification. CONCLUSION: Health workers exposed to biological risk and in particular those who got in contact with infective biological liquids can be considered at risk of neoplasia development; for this reason, we propose a three phases behaviour model: 1. Biological risk accidents prevention; 2. Prevention of infective disease after an accident; 3. Neoplasia prevention and/or early diagnosis after the development of the infective disease. PMID- 18409993 TI - [Latex gloves dermatitis in health care workers]. AB - With regard to health care workers the irritative contact dermatitis represents about the 80% of all the dermopathies in sanitary staff whereas the allergic contact dermatitis covers approximately the 20% of the professional dermatoses. In our study 4 cases of allergy to latex in hospital nurses are presented; the clinical history is described for each of them as well as the resulting judgment of suitability to the specific work. In general population the allergy to latex is estimated to be approximately 1-6%; in sanitary staff it rises to 5-12%. We have to observe that not all the sensitive subjects show symptoms of allergy. Actually the 4 cases described represent less than 1% of the surveyed group. The sensitization is likely to be so reduced also thanks to the application of preventive guide-lines such as the one proposed by NIOSH. In Italy the criteria in preventing allergic reactions to latex are illustrated in a consensus document issued by a study-team from Italian Association of the Health Workers. PMID- 18409994 TI - [Screen dermatitis and visual display units]. AB - Over the last fifteen years, the increasing prevalence of screens and electromagnetic devices used in a number of activities has determined the occurrence of new emerging clinical problems. Hence the need to evaluate the ergonomic aspects of the professions using such equipment to study the effects of electromagnetic fields on human health. The objective difficulties to standardize the studies on this field makes controversial the debate on the evidences in support of these nosographical entities and on the possible causative factors. One interesting area is that of dermatological diseases. The authors present the results of a study on the incidence of dermatological diseases of the face and of the neck in a population of screen workers. To achieve this goal subjects underwent dermatological counselling during the periodic of monitoring visits. Subjects were then identified in two groups: screen workers and as a control group, subjects not exposed to electronic equipments. Members of the first group when compared with those of the second, suffered more frequently from skin diseases, although there was just a statistical trend. However skin diseases were distributed with different frequency in the two groups and seborrheic dermatitis was much more represented among screen workers. PMID- 18409995 TI - [Not allergic dermatitis as occupational diseases: a suggestion for an evaluation method]. AB - Aim of the study was to propose a method to define the relationship of not allergic contact dermatitis with chemical risk factors in workplace. This method evaluates the skin response to simultaneous chemical and mechanical stimulation, made with a soft bristle brush. The method has been used on 9 cases of not allergic skin diseases, observed from 2000 to 2006 in Occupational Medicine Division of "Federico II" University of Naples. The greatest part of pathologies had the polymorphous pattern of skin inflammation, characterized by subintrant erythema, suspected of occupational origin. In 6 cases a stronger reaction to both chemical and mechanical stimulation was highlighted, confirming a possible occupational etiology. In 1 case the occupational origin was excluded, while in 2 cases the skin response was uncertain. The method does not define all cases of occupational skin pathologies, but it seems to be very useful to diagnostic and insurance needs. PMID- 18409996 TI - [Occupational contact dermatitis in metal workers and gender effect]. AB - Contact dermatitis is more frequent among women for anatomical reasons and for extraprofessional exposure to irritants and detergents during homeworks. In addition sensitisation to contact haptens is different in sexes. The aim of our work was to evaluate the association between patch test skin sensitizations and professional exposure to metals analyzing data for gender. Of the 15.217 patients patch tested for dermatitis, 678 were metalworkers. The statistical analysis revealed a significant association between dermatitis and sensitisation to nickel in professional exposed women (OR = 1.68; LC50% 1.11-6.50) while metal sensitisation (Cr.Ni and Co) was not relevant in men: for them a significant association between dermatitis and sensitisation was found to quaternium (OR = 3.91; LC95% 1.18-12.9), to mercaptobenzothiazole (OR = 2.69; LC50% 1.11-6.50) and to ethylendiamine dichloride (OR = 2.53; LC95% 1-6.41). The authors stress the need to evaluate patch test sensitisation considering gender effects. PMID- 18409997 TI - [Nanotubes and occupational medicine]. AB - Carbon nanotubes (CNTs) are a man-made form of carbon, they exist in many different forms and can be chemically modified and/or functionalized with biomolecules. Pristine single-walled CNTs (SWNT) are extremely hydrophobic tubes of hexagonic carbon (graphene) with diameters as small as 0.4 nm and lengths up to micrometers. Multiwalled CNTs (MWCNT) consist of several concentric grapheme tubes and diameters of up to 100 nm. Due to their unique chemical, physical, optical, and magnetic properties, carbon nanotubes have found many uses in industrial products and in the field of nanotechnology, including in nanomedicine. From the rapidly emerging applications of CNTs arise the urgent need for toxicological studies on CNTs. Exposure to CNTs is associated with effect on DNA, pulmonary toxicity, fibrotic lesions, skin toxicity. Very interesting data are collected in a study in which CNTs were seen to connect multiple cells together. Of greatest interest was the discovery of unique intercellular carbon structures composed of SWCNT that bridged lung macrophages. These "carbon bridges" offer a novel and easily identifiable biomarker of exposure. This review brings together toxicological studies that have been carried out using carbon nanotubes, to offer a summary of the state-of-the-art in the development of carbon nanotubes toxicology. PMID- 18409998 TI - [Organic artificial fibers: properties and toxicity]. AB - Now among asbestos substitutes, the man-made organic fibers (MMOFs) find interesting applications on the industrial side. This class includes aramidic, polyacrylic, polyamides, polyolefins and polyvinylic fibers and it has been listed in category 3 of IARC classification (not classifiable as to carcinogenicity to humans). The aim of this study is to provide a panoramic regarding the toxicity of MMOFs present on the market according to their chemical physical properties. Three materials have been taken in to consideration: the polyacrylonitrile (PAN), the polyvinyl alcohol (PVA) and the Kevlar. Each of them has been observed by the Scanning Electron Microscopy and Energy Dispersive Spectroscopy to estimate the morphology and the mean diameter. The obtained mean diameter of fibers is 10.50 microm for the PAN, 16.60 microm for the PVA and 11.61 microm for the Kevlar. The last one presents fibrils having a diameters in the range 0.2-1.6 microm. The ability of the Kevlar to produce fibrils with dimensions less than 1 microm, after abrasion or mechanical stress of other kind, is the cause of numerous studies based on the evaluation of their biosolubility. Finally we show some spectra obtained from the analysis of these fibers through Fourier Transform Infrared Spectroscopy (FTIR). PMID- 18409999 TI - [Relations between health and natural and artificial environment]. AB - On June 2007 WHO issued their first report on impacts of environmental conditions on human health, "Country profiles of the environmental burden of disease". Data relevant to Italy indicate that health diseases caused by environment are 14%. Further papers demonstrated that, in highly technologically developed countries the air available in non industrial buildings (schools, offices, houses, etc.) is more polluted then the air outside and, adding to this that these are the places where population lives up to 90% of the time, this data highlight the importance and extension of the issue. This study recognize, thanks to the cross check of data from various international studies, a strong connection existing between health and psycho physic well-being of human being and a salubrious environment, outdoor and indoor; but, also, the existence of a vicious circle where the outdoor environment degradation lead to buildings indoor pollution and non eco sustainable buildings new erections worsen the outdoor environment. The challenge for the future is to guard natural environment and to achieve buildings suitable for human being needs and demands and in harmony with natural environment. PMID- 18410001 TI - [A workplace intervention aimed at increasing awareness in nursing personnel performing manual handling activities]. AB - Here we describe a workplace intervention aimed at reducing the risk of low back pain in nursing personnel. The intervention we carried out included a specific ergonomic training and an exercise program according to the Feldenkrais Method. After the intervention, we evaluated its effect on the execution of manual handling activities in nurses. We found an increased rate of correct manual handling in the post-intervention period. PMID- 18410000 TI - [Work related sensitization risk in hotel school students, the role of garlic]. AB - We want to evaluate the incidence of allergic problems in a population of catering students at the beginning and after 3 and 5 years of their course. We enrolled in the study 601 students of a professional catering school: 412 in the first year and 189 in the fifth years. We used two questionnaires to investigate respiratory problems and dermatological ones. All the subjects were also patch tested with 10 aptens. The incidence of reported symptoms of allergy was higher in fifth year than in first one. We can observe a rising in positive reaction between the two group of students to Balsam Peru, Nickel, Fragrance mix and Garlic. In particular we found an higher incidence of positive patch reaction in female population. On the contrary the rising in positive reaction to Garlic is higher in male. About two third of the subjects positive to garlic present a sensitization to diallyl disulphide. The rising of positive patch test reactions to Garlic and Balsam Peru should be caused by the contact with cooking spices. The higher incidence of allergic pathology between students during fifth year of school and students at first year is probably due to scholastic practical activity. PMID- 18410002 TI - [Technopathic hearing loss and open fitting]. AB - The purpose of this study is to value the advantage of open fitting ear-moulds in tecnopathic ipoacusies. The study was conducted on a sample of 70 tecnopathic ipoacusics with a hearing loss in the high wave frequencies (4000-8000 Hz) ammounting to at least 50 dB; 20 of which already using ear-moulds. To all of them was applied, for three weeks, an open fitting ear-mould. At the beginning and at the end of the study, the sample was given a questionary to fill out. The analysis of the questionnaries has proven that among the 20 tecnopathics already using ear-moulds: A) 16 subjects have a marked preference for the open fitting ear-moulds, B) 3 did not encounter differences between the two types of ear moulds, C) 1 declared a better adaptability to the previous ear-moulds. Over all, in the 70 subjects examined, this research's results allow us to conclude that: the use of open fitting ear-moulds improves the quality of the natural sound of one's own voice, the natural quality of sounds, and the hearing of speech in the presence of other noises, while reducing irritation of the auditory cortex. PMID- 18410003 TI - [Fireman at work]. AB - The objectives of the research are to identify, through procedures and tools psychosocial investigation, appropriate measures for the improvement of health surveillance and training, in order to promote higher standards of welfare in a population of workers exposed to systematically emergency. For the survey data was used questionnaire "The fireman to work" consists of 52 items, administered anonymously. The areas of the questionnaire include: 1) structural data. 2) labor: organizational (working at the place of residence, age, role, status); 3) interpersonal relationships: interpersonal relations horizontal and vertical (colleagues, employees, above); 4) organizational climate. 5) safety, health and welfare at work. 6) habits and lifestyles. The number of questionnaires returned and the results valid for the analysis was around 50%; Also with a percentage of returns oscillating between 0% of Bologna and 100% of Padua. Therefore it was not possible to consider the questionnaires (3741) as representative of the population in question. However the study is of great importance since is the first survey of its kind carried out in our country. PMID- 18410004 TI - [Air pollution in urban areas as a risk factor for health in the general population and in workers. Environmental and biologic monitoring of 1-3 butadiene]. AB - Exposure to 1,3-butadiene (BD), a probable carcinogen to humans, was investigated in two groups of subjects working in a refinery plant: 28 occupationally exposed workers and 21 subjects non-occupationally exposed controls. BD in blood was higher in the exposed workers than in controls. Excretion of urinary mercapturic acids N-acetyl-S-(3,4-hydroxybutyl)-l-cysteine (MI), was not influenced by occupational exposure. The present study shows that unmetabolised BD in biological fluids, represents the biomarker of choice for assessing occupational exposure to low airborne concentrations of BD. PMID- 18410005 TI - [Application of intraesophageal stent for the treatment of esophageal fistula]. AB - The results of treatment of 52 patients, suffering esophageal fistula, of them 36 -with tumoral origin and 16--due to anastomotic sutures insufficiency, were analyzed. The domestic production stents with partial or complete polytetrafluoroethylene coverage were applied. The stent was put in place under roentgenoscopy guidance according to the authors method. After the stent putting in place for esophagopulmonary fistula, occurred due to esophageal and pulmonary malignancies presence, the patients quality of life have had improved, from the first postoperative day they ingested food per vias naturalis, the respiratory signs severity of food aspiration had lessened, the dyspnea eliminated, the cough lessened and general condition improved. In the patients, suffering postoperative fistula, after the stenting performance, in conditions of anti-inflammatory and desintoxication therapy conduction the esophageal sutures insufficiency signs had subsided or purulent-septic processes have had eliminated, the fistula had closed. PMID- 18410006 TI - [Surgical treatment of biliary fistulas]. AB - The methods of diagnosis and operative treatment of internal (in 19 patients) and external (in 18) biliary fistulas, which appeared due to destructive and complicated forms of cholecystitis were analyzed. PMID- 18410007 TI - [Differentiated approach to surgical treatment of inguinal hernia recurrence in accordance with morphologic characteristics of musculo-aponeurotic structures of inguinal channel]. AB - In 44 patients, ageing 39-73 yrs and suffering inguinal hernia recurrence, the investigation of musculo-aponeurotic structures of inguinal region was conducted. In 21 patients, suffering primary inguinal hernia recurrence, in concurrence with the connective tissue complex degeneration the pronounced atrophy of tissues was revealed. In 23 patients, suffering multiple inguinal hernia recurrence, the tissues degeneration and atrophy were manifested significantly. The results of investigation trust the necessity of additional plastic material application while performing hernioplasty for inguinal hernia recurrence. To optimize the primary inguinal hernia recurrence, the combined hernioplasty, applying polypropylene mesh, was performed, and for secondary recurrence--preperitoneal allohernioplasty, using operative access through the inguinal channel. PMID- 18410008 TI - [Treatment of patients with three-atrium heart]. AB - Typical three-atrium heart constitute a rare inborn heart failure, in which pulmonary veins drain into the proximal additional camera of the left atrium, separated from distal left pericardium cavity of it own by diaphragm, in which there is one or more restrictive apertures. Of 6770 patients, consecutively operated on by one surgeon, in 15 (0.21%) ageing from 7 mo to 30 years the three atrium heart was revealed. In all the patients the surgical correction of the failure was accomplished using artificial blood circulation. The early diagnosis and surgical treatment of the patients, suffering three-atrium heart failure, secures good late results and quality of life. PMID- 18410009 TI - [Local hemostasis during the carotid endarterectomy under condition of antithrombocytic therapy]. AB - Comparative studying of preparation TachoComb application efficacy and the convenient tampon procedure for stopping of bleeding from the suture line, fixing the expanding flap, made of polytetrafluoroethylene, in arteriotomy zone was studied in the patients, operated for carotid arteries stenosis and obtaining Plavix preparation protractedly in 75 mg dosage. The preparation TachoComb application have had promoted the operative time reduction as well as the bleeding duration and the blood loss volume. PMID- 18410010 TI - [The changes of immunoreactivity in patients suffering polynodose euthyroid goiter and their influence on the results of surgical treatment]. AB - The immunity changes were studied preoperatively in 36 patients suffering polynodose euthyroid goiter and in 85--postoperatively. The immunity system disorders are more profound in the early terms after the operation and their severity directly depends on its volume. The immunity changes are significantly expressed in adenomatous transformation of the thyroid gland tissues and in postoperative hypothyroidism. PMID- 18410012 TI - [The model of gastroesophageal reflux disease]. AB - Elaboration and clinical application of new methods of treatment is impossible without their all-round approbation in experiment. It necessitates the models creation, which, due to their pathogenetic mechanisms, would be maximally adopted to clinical conditions. The experimental model of gastroesophageal reflux disease, application of which permit to eliminate the antireflux defense factors and to create the conditions for gastroesophageal reflux studying in chronic experiment, was created. PMID- 18410011 TI - [Rational application of the pneumatic tourniquet in the knee joint endoprosthesis]. AB - Two different methods of the pneumatic tourniquet application were used while the knee joint prosthesis in two groups of patients: during the all operation and during the stage of implantation of artificial joint components. It is preferent to reduce the pneumatic tourniquet application duration to realize its positive qualities, to secure optimal adhesion of cement and costal bed and also to make less severe the consequences of intra- and postoperative period. The data obtained trust that the procedure, used by us, constitutes an optimal method of the pneumatic tourniquet application while the knee joint cement prosthesis. PMID- 18410013 TI - [Restorative surgery in large intestine cancer]. PMID- 18410014 TI - [Venous thrombosis of the lower extremities. Is it possible to solve the problem today?]. PMID- 18410015 TI - [Foreign bodies of the large intestine]. PMID- 18410016 TI - [Observation of unusual cardiac lipoma]. PMID- 18410017 TI - [Three patients with tuberculous meningitis: treatment started at tentative diagnosis]. AB - A 30-year-old man and a 37-year-old woman with no history of tuberculosis developed symptoms of headache, vomiting and subsequent aggressive behaviour. After several lumbar punctures, the PCR test for tuberculosis in the cerebrospinal fluid was positive, and a definitive diagnosis of tuberculous meningitis was made. Treatment with antimycobacterial agents was not started until a few days after hospital admission. The man recovered, but was treated for brainstem tuberculoma 12 months later; the woman died on day 11 of hospitalisation. A third patient, a 31-year-old man, was admitted to the hospital for miliary tuberculosis. He had signs of progressive apathy and meningismus. Mycobacterium tuberculosis was found in his cerebrospinal fluid. Each of these patients underwent cerebrospinal fluid drainage due to communicating hydrocephalus and each had hyponatraemia. Tuberculous meningitis is a lethal complication of tuberculosis that is often diagnosed late due to the insidious nature of its symptoms. Early treatment with antituberculous drugs and dexamethasone--even before a definitive microbiological diagnosis is made--may prevent severe neurological damage and death. PMID- 18410018 TI - [Prognosis in patients operated on for colorectal carcinoma: related to but not determined by number of lymph nodes examined]. AB - The presence or absence of lymph node metastases is pivotal for the estimation of prognosis in carcinoma patients. The presence of lymph node metastases in patients with colorectal carcinoma (CRC) is an indicator of poor prognosis and adjuvant treatment is increasingly being given to these patients. For the patient much depends on effective staging of CRC. In patients with TNM stage II CRC, prognosis is related to the actual number of nodes harvested: those in whom fewer nodes are harvested fare less well. Understaging was suspected and therefore protocols now require a minimum number of nodes. Recent observational studies, however, have questioned understaging as a plausible mechanism. Tumour, biological or patient-related factors seem to be more likely. The number of nodes is not a suitable measure for the quality of pathological staging of CRC. Instead of this outcome measure, more attention should be paid to processes and future standards must be based on sound experimental evidence and not on observational surveys. PMID- 18410019 TI - [Perinatal mortality in The Netherlands: an audit is now more necessary than ever]. AB - In 1999, The Netherlands was found to have the highest perinatal mortality rate (11.4 per thousand), according to the WHO definition, in comparison with 14 other European countries. Many explanations were given, amounting essentially to a higher percentage of women with one or more risk factors (higher age at delivery, more non-western immigrants, and more multiple pregnancies). Furthermore, screening for congenital anomalies was not yet an issue at that time in The Netherlands. Finally, Dutch obstetricians and paediatricians have a restrictive policy regarding the treatment of newborns with a very poor prognosis. There are, however, also doubts about the quality of the delivered perinatal care. In an audit feasibility study in 2004 in a representative part of The Netherlands, it was found that in 9% of the cases of perinatal mortality, substandard care was (very) probably related to the mortality. In The Netherlands, a nationwide perinatal audit programme will start this year. The Netherlands, with its specific obstetric care system, has the obligation to look for the causes of perinatal mortality and to introduce improvements based on the results of a perinatal audit. Hasty conclusions are a threat to a careful audit. PMID- 18410020 TI - [Clinical reasoning and decision-making in practice. A patient with fever and pancytopenia]. AB - An 82-year-old man was admitted with a 1-week history of chilling fever and dry cough. Laboratory tests revealed pancytopenia and elevated levels of C-reactive protein and lactic dehydrogenase (LDH). Screening for infectious diseases was negative. A bone marrow biopsy showed aspecific findings. The combination of pancytopenia, persistent fever, elevated LDH and hepatomegaly (demonstrated by ultrasound examination of the abdomen) was suggestive of the haemophagocytic syndrome. This was confirmed by very high levels of ferritin and soluble interleukin-2 receptor in the blood. In addition, re-examination of the bone marrow showed several haemophagocytic histiocytes. A polymerase chain reaction for Epstein-Barr virus (EBV) revealed a very high viral load. Since the patient had a history of an increased level of anti-EBV immunoglobulin-G, this was explained by a reactivation of the EBV infection. On the sixth day in hospital the patient developed signs of bilateral pneumonia and subsequent multiple organ failure. Despite intensive treatment the patient died. Autopsy revealed no haematological or other malignancies, but did show haemophagocytosis in many organs. It was then concluded that the patient had a virus-associated haemophagocytic syndrome, due to a reactivation of EBV, for which no underlying cause was found. PMID- 18410021 TI - [Diagnostic image (365). A man with facial hyperpigmentation]. AB - A 79-year-old man developed a blue-greyish facial hyperpigmentation after amiodarone administration over 4 years. PMID- 18410022 TI - [Estimation of the incidence of tuberculosis in The Netherlands in 2030]. AB - OBJECTIVE: To estimate the number of cases of tuberculosis (TB) in 2030 for the purpose of planning future TB control. DESIGN: Statistical modelling in 5-year intervals until 2030. METHOD: The number of Dutch TB cases infected by a Dutch source was estimated using a survival model. The number ofnon-Dutch patients was estimated by calculating the proportion of culture-positive TB patients among first-generation immigrants in 2005 and applying this proportion to the projected size of the non-Dutch population. It was assumed that each non-Dutch TB patient infected by a non-Dutch source would cause one infection in the population in The Netherlands. RESULTS: The estimated number of TB cases is expected to decrease to 877 in 2010. Only a limited decrease in the number of TB patients is expected after 2010 as the number of non-Dutch TB cases increases due to increased immigration. This increase negates the expected decrease in Dutch TB patients infected by a Dutch source. In 2030, non-Dutch TB cases will account for 85% of all TB cases. The proportion of non-Dutch TB cases is greater in the 4 largest cities, i.e. Amsterdam, Rotterdam, The Hague and Utrecht (89%) than in the rest of The Netherlands (76%). CONCLUSION: The decrease in TB incidence observed over the past several years may cease by 2010 due to an increase in non-Dutch TB patients as a result of increased immigration. However, the confidence intervals associated with these estimates were large. Future TB control efforts must be organised in a flexible way so that they can be adapted to changing epidemiological situations. PMID- 18410023 TI - [Characteristics and treatment of tuberculosis patients in Dekkerswald, 2000 2005]. AB - OBJECTIVE: To describe the patient population in Dekkerswald, Nijmegen, one of two tuberculosis (TB) centres in The Netherlands. DESIGN: Descriptive, retrospective study. METHOD: Examination of medical records for all TB patients hospitalised between 2000 and 2005, including demographic, social, clinical and follow-up data. RESULTS: Data from 166 patients were analysed. Tertiary referrals accounted for 98% of all hospitalisations. Most patients (68%) were referred for clinical reasons, and 32% were referred for social reasons. Drug resistance was encountered in 23% of patients; 9% had multidrug-resistant TB. Ten percent of patients were seropositive for HIV. Toxicity and side-effects of treatment often led to changes in treatment (40%). Patients had pulmonary TB (59%), extrapulmonary TB (23%) or both (17%). Overall, 141 patients (85%) completed treatment. The TB-related mortality rate was 5%. CONCLUSION: In Dekkerswald, there is a selected patient population that is characterised by drug-resistance, comorbidity, side-effects, extrapulmonary disease and social issues. Due to the low prevalence of TB in The Netherlands, knowledge and experience regarding complex types of TB are limited. Centralisation of patient care is important to preserve and optimise this expertise. PMID- 18410024 TI - [A Moroccan woman with genital tuberculosis]. AB - A 28-year-old nulliparous Moroccan woman presented at an outpatient gynaecology clinic because she had been unable to conceive for the last 2 years. She had an enlarged right adnexus. Histopathological examination of the resected right adnexus was suspicious for tuberculosis. After isolating Mycobacterium bovis from a psoas abscess 3 months later, a 6-month regimen with anti-tuberculosis drugs was started, and the clinical response was good. Female genital tuberculosis is rare in developed countries. Symptoms are few and non-specific. The disease can be suspected incidentally during the subfertility work-up. The fallopian tubes are the most commonly affected site. A definitive diagnosis is difficult to make, even when tuberculosis is strongly suspected. Histopathological findings can support a diagnosis of tuberculosis, and molecular PCR techniques that detect DNA are promising. Treatment of genital tuberculosis is similar to that of pulmonary tuberculosis, although it is difficult to monitor treatment response. The chance of spontaneous conception after treatment is very small, and IVF is often the only treatment option. PMID- 18410025 TI - [Bilateral cataract in childhood years: always an indication for screening on a metabolic disorder]. AB - In three young patients who presented with bilateral cataracts the cause proved to be an inherited metabolic disease. The first patient was a newborn aged 7 weeks, in whom galactokinase deficiency was diagnosed. The second patient was a boy aged 8 years with cerebrotendinous xanthomatosis. The third patient was a girl who was diagnosed with cataracts at the age of 3 months. At the age of 4 years the diagnosis 'rhizomelic chondrodysplasia punctata' was established. Screening for metabolic disorders in all children with bilateral cataracts is essential, as in some disorders progressive and severe symptoms can be avoided with timely initiation of treatment. In addition, diagnosis allows for family studies and genetic counselling to take place. This may result in prevention of disease by early therapeutic intervention and prenatal screening. PMID- 18410026 TI - [Increase in extrapulmonary tuberculosis in The Netherlands associated with an increase in the number of residents with non-Dutch nationality; observational study of data from 1993-2001]. AB - OBJECTIVE: To describe the epidemiology of extrapulmonary and pulmonary tuberculosis (TB) in The Netherlands between 1993 and 2001, with special attention given to the relationship between disease site and patient nationality. DESIGN: Retrospective descriptive and comparative study. METHOD: Data were collected from all patients diagnosed with TB and entered in The Netherlands Tuberculosis Register between 1 January 1993 and 31 December 2001. Information on age, sex, nationality, year of diagnosis, culture result, disease site and HIV status was retrieved and compared. RESULTS: Of the total 13,258 TB patients, 8,216 (62%) had pulmonary TB and 5,042 (38%) had extrapulmonary TB. The proportion of extrapulmonary TB cases increased during the study period. The most common types of extrapulmonary TB were lymphatic TB, pleural TB and TB of the bones and/or joints. Patients with a non-Dutch nationality were diagnosed with extrapulmonary TB more often than Dutch patients. CONCLUSION: The proportional increase in extrapulmonary TB was explained by an increase in the number of inhabitants with non-Western ethnic backgrounds in The Netherlands. Physicians must be aware of the changing clinical picture of TB. PMID- 18410027 TI - [Dutch study on the optimal treatment strategy for patients with a first or second occurrence of gastro-oesophageal variceal bleeding: the TIPS-TRUE trial]. AB - The main options for secondary prevention of gastrooesophageal variceal bleeding are endoscopic therapy and treatment with propranolol. Creation ofa transjugular intrahepatic portosystemic shunt (TIPS) is currently considered a valuable secondary 'rescue' treatment when other therapies fail. Recent data suggest that the use of covered stents markedly increases the efficacy of TIPS, compared with conventional uncovered stents. Therefore, a multicentre randomised trial was designed to compare the effects of TIPS using covered stents with those of endoscopic therapy plus propranolol in patients with a first or second episode ofgastro-oesophageal variceal bleeding. TIPS will be performed in 4 university centres with relevant expertise. The trial will hopefully gain nationwide support, and all centres in The Netherlands are cordially invited to participate. PMID- 18410028 TI - [Theo Ooms]. PMID- 18410029 TI - [AIDS treatment in Africa]. PMID- 18410030 TI - [Overweight and mortality risk: no connection between overweight in middle and older age groups and increased mortality]. PMID- 18410031 TI - [Axial length of the eye in the first three years of life in children with congenital cataract in one or both eyes]. AB - INTRODUCTION: Ultrasound has an important role in the diagnostics of the posterior segment of the eye, when clinical examination is not possible. One of problems is congenital cataract. Using ultrasound biometry it is possible to measure the axial length of the eye in all ages and axial growth from the very beginning to its termination. OBJECTIVE: To ratify growth of the infant's eye with and without congenital cataract, and to determine the accuracy of ultrasound measuring with and without general anaesthesia. METHOD: In our study we analyzed 30 patients (48 eyes) who had congenital cataract in one or both eyes (observed group) and 20 patients (40 eyes) in whom no eye disease was diagnosed (control group). RESULTS: The observed group of 30 patients (48 eyes) with congenital cataract was divided according to age into three subgroups: 1) 4-12 months, 11 patients, 18 eyes; II) 13-24 months, 7 patients, 11 eyes; III) 25-36 months, 12 patients, 19 eyes. In control group we analyzed 20 patients (40 eyes) without congenital cataract. These patients were also divided, according to age, into three subgroups: 1) 2-12 months, 5 patients, 10 eyes; II) 13-24 months, 6 patients, 12 eyes; III) 25-36 months, 9 patients, 18 eyes. CONCLUSION: Comparing the average axial lengths of the eye in the first, second and third year of life between the observed and control groups, we did not find any statistical significance in the obtained values. When determining the accuracy of the ultrasound measuring of the axial length of the eye in the observed group, we found no statistical significance between the determined values weather the measurements were done with or without general anaesthesia (p>0.05). PMID- 18410032 TI - [Sub-tenon anaesthesia in the operation of cataract using phacoemulsification technique]. AB - INTRODUCTION: Already in 1884, Knapp reported a successful cataract operation using 5% of cocaine in eyedrops.Today, different methods of local anaesthesia in the operation of cataract are used, such as retrobulbary, peribulbary, subconjunctival, sub-tenon, topical and intracameral anaesthesia. Phacoemulsification is a modern approach to the operation of cataract that is increasingly applied in our country. Aiming to eliminate the risk of severe complications caused by retrobulbary anaesthesia, in our patients undergoing surgery to treat cataract we applied sub-tenon anaesthesia. OBJECTIVE: The aim of this work was to establish the efficacy and the safety of subtenal anaesthesia in the operation of cataract using phacoemulsification technique. METHOD: We examined 85 patients who were operated for cataract under sub-tenon anaesthesia. The operation was performed using phacoemulsification technique. The surgeon injected about 2 ml of the mixture of lidocaine (2%) and bupivacaine (0.5%) in equal proportions into the bottom of the fornix at 11 o'clock or 1 o'clock position into one of the two upper eye quadrants. After phacoemulsification and the placement of intraocular lenses, the patients were asked to reply to the questions according to a standardized protocol. Priory, the patients were informed about the test and the pain scale ranging from 1 to 10, with 1 indicating the absence of pain and 10 unbearable pain. RESULTS: Eighty-five patients underwent surgery for cataract using phacoemulsification technique and in whom posterior chamber intraocular lenses were placed under sub-tenon anaesthesia. During the application of anaesthesia, subjective pain experience was at the level of mild discomfort or non-existent in 3/4 of the patients. In relation to pain experience during surgery, it ranged from mild inconvenience to pain absence in 4/5 of the patients. The most frequent complication during surgery was eye rolling occurring in every 10th patient. In 4/5 patients the overall surgical course was assessed by the surgeon as excellent or outstanding. CONCLUSION: Sub-tenon anaesthesia applied at one point of the upper quadrant using during modern approach in the surgery of cataract, namely the phacoemulsification technique, offers good comfort to the patient during anaesthesia introduction, as well as during surgery, with a relatively low complication rate occurring during the operation, and also offering good comfort to the surgeon. PMID- 18410033 TI - [Prognostic value of QTc interval dispersion changes during exercise testing in hypertensive men]. AB - INTRODUCTION: The prognostic significance of QTc dispersion changes during exercise testing (ET) in patients with left ventricular hypertrophy is not clear. OBJECTIVE: The aim was to study the dynamics of QTc interval dispersion (QTcd) in patients (pts) with left ventricular hypertrophy (LVH) during the exercise testing and its prognostic significance. METHOD: In the study we included 55 men (aged 53 years) with hypertensive left ventricular hypertrophy and a negative ET (LVH group), 20 men (aged 58 years) with a positive ET and 20 healthy men (aged 55 years). There was no statistically significant difference in the left ventricular mass index (LVMI) between LVH group and ILVH group (160.9+/-14.9 g/m2 and 152.8+/-22.7 g/m2). The first ECG was done before the ET and the second one was done during the first minute of recovery, with calculation of QTc dispersion. The patients were followed during five years for new cardiovascular events. RESULTS: During the ET, the QTcd significantly increased in LVH group (56.8+/ 18.0 - 76.7+/-22.6 ms; p<0.001). A statistically significant correlation was found between the amount of ST segment depression at the end of ETand QTc dispersion at the beginning and at the end of ET (r=0.673 and r=0.698; p<0.01). The QTc dispersion was increased in 35 (63.6%) patients and decreased in 20 (36.4%) patients during the ET. Three patients (5.4%) in the first group had adverse cardiovascular events during the five-year follow-up. A multiple stepwise regression model was formed by including age, LVMI, QTc interval, QTc dispersion and change of QTc dispersion during the ET. There was no prognostic significance of QTc interval and QTc dispersion during five-year follow-up in regard to adverse cardiovascular events, but prognostic value was found for LVMI (coefficient beta=0.480; p<0.001). CONCLUSION: The increase of QTc interval dispersion is common in men with positive ET for myocardial ischemia and there is a correlation between QTc dispersion and amount of ST segment depression. The increase of QTc interval dispersion during negative ET in patients with left ventricular hypertrophy has no significant predictive value for adverse cardiovascular events, as shown in our five-year follow-up study, but it requires confirmation by investigation of larger groups of patients. PMID- 18410034 TI - [Determinants of body mass index in children and adolescents]. AB - INTRODUCTION: Body Mass Index (BMI) in boys and girls is predicted by parental BMI, age and occupation. OBJECTIVE: Correlation of BMI among children and adolescents in Banjaluka region (Bosnia and Herzegovina) and parental age, BMI, parents'educational level and occupation, as well as the number of family members were investigated as the possible determinants of overweight and obesity in childhood. METHOD: The study included 1204 children and adolescents (578 males, 626 females), 6-17 years old from primary and secondary schools in the Banjaluka region. BMI was calculated from height and weight using the standard formula. Each subject along with his parents answered the questionnaire that contained information about parents' height and weight, educational level and occupation, as well as the number of family members. RESULTS: In all studied children, the prevalence of overweight was 12.2% and of obesity 6.1%. Strong positive correlation was found between parental BMI and age (older than 40 years) in males and females (p<0.001), while parental higher BMI and higher educational level had positive correlation only in males (p<0.001). The number of family members showed negative correlation with overweight/obesity only in females. CONCLUSION: The prevalence of overweight and obesity in children's population in the Banjaluka region is 12.2% and 6.1%, respectively. There is a positive correlation of overweight and obesity in children with parental overweight and obesity, as well as older age, and parental higher educational level. PMID- 18410035 TI - [Validity of clinical and ultrasound examination related to arthroscopy in acute injury of the medial meniscus of the knee]. AB - INTRODUCTION: Injuries of the medial meniscus of the knee are constantly present in sports, but also in common life activities. Diagnostic procedures which are available today in the diagnosis and estimation of the severity of intraarticular knee structures are numerous, but they are not available, plausible and valid at the same manner. OBJECTIVE: The aim of this study was to compare the diagnostic validity of clinical and ultrasound examination related to arthroscopy in acute injury of the medial meniscus of the knee. METHOD: This prospective study analyzed 198 hospitalised patients, treated from 2004 to 2006 at the Belgrade Institute for Orthopaedic Surgery "Banjica" and Belgrade Special Orthopaedic Surgery Hospital "Decedra", who underwent arthroscopy. Before the arthroscopy, clinical and ultrasound examinations were performed. RESULTS: The values of sensitivity (91.1%), specificity (80.0%), positive predictive value (83.6%) and negative predictive value (88.9%) of ultrasound examination of acute injury of the medial meniscus are proportionately lower as compared to the values of sensitivity (97.2%), specificity (90.2%), positive predictive value (86.0%) and negative predictive value (98.2%) of ultrasound examination in chronic injuries of the medial meniscus of the knee. CONCLUSION: Ultrasound examination is a more useful method in the detection of lesions of the medial meniscus of the knee as related to clinical tests used in their detection in this study. PMID- 18410036 TI - [The significance of age and sex for the absence of immune response to hepatitis B vaccination]. AB - INTRODUCTION: Seroepidemiological investigations after the administration of hepatitis B vaccine have shown that even 15% of vaccinated healthy persons do not generate immune response to the vaccines currently in use. OBJECTIVE: The aim of the research is to test the immunogenicity of hepatitis B vaccine in different age groups on the adult vaccinated population sample in Serbia. METHOD: The tested general population sample consisted of 154 adult subjects. Immunization was done using the recombinant fungal vaccine obtained by genetic engineering (Euvax B vaccine, manufacturer LG, distributor Sanofi Pasteur). All tested subjects in the research received 1 ml of hepatitis B vaccine administered intramuscularly into the deltoid muscle by 0, 1, 6 schedule. RESULTS: In the tested sample, 3.13% of persons aged up to 29 years, 6.25% aged 30-35 year and 19.23% of the tested persons aged 40 years and older had no immune response. The relative risk of"no response"findings was twice higher in the group aged 30-39 as compared to the population aged up to 29 years. The detected risk was six times higher for the population of 40 years and older in comparison to the population aged up to 29 years. Also, the relative risk of"no response" findings for the population of 40 years and older was more than three times higher than for the group aged 30-39. Absent immune response in relation to sex was found to be higher in male subjects. CONCLUSION: The rates of"no response"finding was the following: 3.13% in the group aged up to 29 years, 6.25% in the group aged 30-39, as well as in the group aged 40 years and older (19.23%). Immune response in relation to age groups was statistically significantly different (p<0.001), while there was a statistically significant correlation (C=0.473; p<0.001) between the age of the subjects and the immune response. In relation to sex, the "no response"finding was found to be increased in the males, but without any statistically significant difference (p>0.05). PMID- 18410037 TI - [Face identification by computerized superposition method: methodology and two case reports]. AB - INTRODUCTION: Skull-face photograph superposition is one of the methods of identification. Digitally recorded and stored within the computer, the images of the skull and face could be superimposed on the monitor. The method requires cooperation among the anthropologist, odontologist, forensic pathologist and the computer technician so as to avoid objective and subjective errors in the identification. CASE OUTLINES: We present two cases of positive identification by superimposition. In the first case, it was a 65 year-old male with several brain operations, thus surgical skull bone-trepanations could be seen as the irregularities on the forehead, and were used as the antropological identificational figure. In the second case, it was an 83 year-old female, whose positive identification was made according a photograph taken at least 26 years before death. CONCLUSION: Face identification by computerized superposition method is usefull in all cases where both the skull and the photograph of the missing person are present, and where other methods of identification have failed due to multiple reasons. PMID- 18410038 TI - [Incindentally detected Castleman disease in a patient with allergic rhinosinusitis]. AB - INTRODUCTION: Castleman disease was for the first time described in 1956 as a mediastinal tumour mass. Etiology of this disease is still unknown. The disease can be solitary and multicentric or rarely of a mixed type. The former is often of hyaline vascular type, while the latter is of plasma cell type. CASE REPORT: Castleman disease was diagnosed in a 26-year old male patient when a well defined shadow was incidentally detected in the middle lobe of the right lung. A year before, he was diagnosed with allergic rhinitis to Ambrosia. Two years after surgery the patient was feeling well, and was without any recurrence, however, allergic rhinitis still persisted. CONCLUSION: Castleman disease can occur in any organ containing lymph tissues. Most frequently the disease is described as mediastinal, rarely as an intrapulmonary tumorous mass, and it is most frequently seen in younger persons. The solitary type of Castleman disease is surgical treatable with a prospect of good prognosis, while the multicentric and mixed types recur despite treatment with cortisone, irradiation and cytostatics. As the association between Castleman disease and allergic diseases has not been confirmed up-to-now, it could be concluded that this patient suffered from two separated diseases. PMID- 18410039 TI - Shoulder joint pathology--improved diagnosis by magnetic resonance imaging (MRI): a pictorial essay and review. AB - This paper describes the personal experience of the author with Shoulder Arthro MR. This imaging method is put into perspective with other, more widely used diagnostic methods, which also have their advantages. Indications for Shoulder Arthro MR are discussed and details on the technique and interpretaton are given. Also major shoulder pathology as impingement, rotator cuff tears and instability is discussed. Furthermore a review of the spectrum of pathology in 100 consecutively examined patients is made and treatment and outcome are listed. Lastly a pictorial review of typical pathologies in MR pictures is supplemented. After x-ray, Shoulder MR is now considered the standard among the imaging methods to diagnose shoulder disorders, as it allows for optimal depiction of pathology and diagnosis in all major imaging planes. At the same time the method gives the arthroscopist an excellent roadmap for his intervention. All in all, the treatment outcome of shoulder trauma has been improved by shoulder MR in conjunction with modern surgical and arthroscopic therapies. PMID- 18410040 TI - [Problems with current hypertension definition in children]. AB - The Task Force data have added immeasurably to our understanding of the normal distribution of blood pressure in children. However, the manner in which arterial hypertension is defined in children is not without problems. The main problem is that the current definition of hypertension in children is not based on the end organ damage assessment, but on the blood pressure height-related percentile distribution of healthy reference population. This could be overcome by introducing the relationship of blood pressure values with sensitive markers of hypertensive sequelae (such as carotid intimal-medial thickness, left ventricle mass index, retinal arteriolar narrowing and arteriovenous nicking as well as microalbumin urinary excretion) to define better the specific blood pressure values with outcomes. Furthermore, the current definition of hypertension based on the demographic and clinical characteristics of the reference population does not consider the normal evolution of reference population, as well as its ethnic differences. In addition, being based on the single occasion measurement in quite position it does not account for the possibility of transient, stress induced elevations in blood pressure known as white coat hypertension. Therefore, incorporation ambulatory 24 h blood pressure data into the definition of arterial hypertension in children may increase the definition reliability for clinical decision - making, although for such reliability the paediatric normative ambulatory blood pressure data should be improved. PMID- 18410041 TI - [Hipoccrates and psychiatry]. AB - Hippocrates, the "father of medicine", is the establisher of a new scientific approach in medicine. He has followed the Hellenic philosophical school that man is the measure of all things, so the "cult of health" is a part of the essence of life. He thoroughly systemized all parts of medicine. He is the establisher of positive (inductive) medicine, which throws away superstition. His approach to medicine is as that of art and natural sciences, and not only as a skill. In his teaching he uses dialectic principles, the establish concept of knowing aetiology, systematization of diagnostics and therapy. He underlined great importance of prognoses of disease based on empirics. He is the founder of holistic medicine, modern concept of psychophysiology, which significantly enabled the disclosure of the etiopathogenesis of psychiatric disorders. His teaching is persistently psychosomatic and weaves in the most modern concepts of psychiatry today. In his aphorisms he described diagnostics and enrolled the principals of treatment of different kind of psychiatry disorders as depression, mania, hysteria, neurotic disorders, psychosis and psychosomatic diseases. PMID- 18410042 TI - Why do we need medical writers, editors and translators? AB - Effective communication in the healthcare sector is imperative, especially nowadays in English. Many scientists and physicians in hospitals, universities and industry either do not have the time or the inclination to write about and publish their research. Medical writers, editors and translators offer a unique service as members of interdisciplinary teams in the drug approval process and medical and scientific communications. PMID- 18410043 TI - Recognition and identification of UV-absorbing leachables in EPREX pre-filled syringes: an unexpected occurrence at a formulation-component interface. AB - During the period of 1998 to 2002, there was an increase in the incidence of antibody-positive pure red cell aplasia (PRCA) in patients receiving subcutaneous administration of EPREX (epoetinum alfa). As part of the investigation of this event, the aqueous formulation containing polysorbate 80, introduced in 1998, facilitated the leaching of small-molecule, aromatic compounds from the uncoated rubber syringe stoppers. The leachables were identified using Liquid Chromatography-Mass Spectroscopy, Electrospray Ionisation-MS/MS, Dithiothreitol reduction, and Hydrogen/Deuterium exchange. The major leachable was identified as a dialkylphenol disulfide, and the majority of the remaining peaks were identified as structural variants containing different numbers of sulfur atoms in the sulfide bridge. In this report, we describe the strategies and experimental designs that were used to overcome the analytical challenges and that led to successful structural identification of the leachables in EPREX pre-filled syringes with uncoated syringe stoppers. PMID- 18410044 TI - Homogenization technique for analysis of post-iontophoretic acyclovir content in porcine skin. AB - The aim of present paper is to develop a method of analysis of acyclovir in porcine skin after iontophoretic delivery using a novel homogenization technique. There are various reported methods available for the analysis of acyclovir in skin samples. All of these methods involve assumption of complete absorption of spiked drug by skin slices for validation. In the current work this assumption has been removed. The method utilizes a step of homogenization to convert the skin into plasma-like matrix, which can be then conveniently used for validation of method as it is done for any plasma sample. It was found that this method can be conveniently used for the estimation of acyclovir in skin layers and that it also virtually removes the step of extraction. PMID- 18410045 TI - Mechanistic studies of glass vial breakage for frozen formulations. I. Vial breakage caused by crystallizable excipient mannitol. AB - The process of freeze-thaw not only subjects bioproducts to potentially destabilizing stress, but also imposes challenges to retain container integrity. Shipment and storage of frozen products in glass vials and thawing of the vials prior to use at clinics is a common situation. Vial integrity failure during freeze-thaw results in product loss and safety issues. Formulations of biomolecules often include crystallizable excipients, which can cause glass vial breakage during freeze-thaw operations. In this study, mannitol formulations served as models for mechanistic investigation of root causes for vial breakage. Several parameters and their impacts on vial breakage were investigated, including mannitol concentration (5% and 15%), different freeze-thaw conditions (fast, slow, and staging), fill configurations (varying fill volume/vial size ratio), and vial tray materials (plastic, stainless steel, corrugated cardboard, aluminum, and polyurethane foam). The results in this study were subjected to a statistical proportion test. The data showed that large fill volumes strongly correlated with higher percentage of vial cracks. Furthermore, the 15% mannitol was found to cause more breakage than 5% mannitol, especially with fast temperature gradient. Significantly more thawing vial breakage occurred in the fast compared to slow freeze-thaw with all types of vial trays. The freezing breakage was substantially lower than the thawing breakage using the fast temperature gradient, and the trend was reversed with the slow temperature gradient. An intermediate hold at -30 degrees C prior to further decrease in temperature proved to be a practical approach to minimize mannitol-induced vial breakage. Thermal mechanical analysis (TMA) and strain gage techniques were employed to gain mechanistic insights, and it was found that the primary causes for mannitol-induced vial breakage were partial crystallization during freezing and "secondary" crystallization of non-crystallized fraction during thawing. The strain on the vial's axial direction was significantly higher than the hoop direction, typically resulting in bottom lens of the vial coming off. Without a 30 degrees C hold, rapid volume expansions due to initial crystallization and secondary crystallization of mannitol were observed in TMA profiles, and these expansions were more apparent in 15% mannitol compared to 5% mannitol. With the introduction of a -30 degrees C hold step, abrupt expansions diminished in TMA profiles, suggesting that most of the mannitol crystallization occurred concurrently with ice solidification during the -30 degrees C holding step and, thus, secondary crystallization during thawing was minimal and the sudden expansion event was eliminated. Therefore, vial breakage during both freezing and thawing was reduced. PMID- 18410046 TI - Mechanistic studies of glass vial breakage for frozen formulations. II. Vial breakage caused by amorphous protein formulations. AB - In an accompanying article we have described parameters that influence vial breakage in freeze-thaw operations when using crystalizable mannitol formulations, and further provided a practical approach to minimize the breakage in manufacturing settings. Using two diagnostic tools-thermal mechanical analysis (TMA) and strain gage, we investigated the mechanism of mannitol vial breakage and concluded that the breakage is related to sudden volume expansions in the frozen plug due to crystallization events. Glass vial breakage has also been observed with a number of frozen protein formulations consisting of only amorphous ingredients. Therefore, in this study, we applied the methodologies and learnings from the prior investigation to further explore the mechanism of vial breakage during freeze-thaw of amorphous protein products. It was found that temperature is a critical factor, as breakage typically occurred when the products were frozen to -70 degrees C, while freezing only to -30 degrees C resulted in negligible breakage. When freezing to -70 degrees C, increased protein concentration and higher fill volume induced more vial breakage, and the breakage occurred mostly during freezing. In contrast to the previous findings for crystallizable formulations, an intermediate staging step at -30 degrees C did not reduce breakage for amorphous protein formulations, and even slightly increased the breakage rate. The TMA profiles revealed substantially higher thermal contraction of frozen protein formulations when freezing below -30 degrees C, as compared to glass. Such thermal contraction of frozen protein formulations caused inward deformation of glass and subsequent rapid movement of glass when the frozen plug separates from the vial. Increasing protein concentration caused more significant inward glass deformation, and therefore a higher level of potential energy was released during the separation between the glass and frozen formulation, causing higher breakage rates. The thermal expansion during thawing generated moderate positive strain on glass and explained the thaw breakage occasionally observed. The mechanism of vial breakage during freeze-thaw of amorphous protein formulations is different compared to crystallizable formulations, and accordingly requires different approaches to reduce vial breakage in manufacturing. Storing and shipping at no lower than -30 degrees C effectively prevents breakage of amorphous protein solutions. If lower temperature such as -70 degrees C is unavoidable, the risk of breakage can be reduced by lowering fill volume. PMID- 18410047 TI - Physicochemical properties of lipid emulsions formulated with high-load all-trans retinoic acid. AB - The objective of this study was to formulate high loading with good stability of all-trans retinoic acid (ATRA) lipid emulsion. Lipid emulsions loaded with ATRA were composed of lecithin, medium chain triglyceride and poloxamer-188 or polysorbate-80. The formulation factors in a particular type and amount of oil, emulsifier, and co-emulsifier on the physicochemical properties (i.e., particle size, size distribution, droplets surface charge, pH, percentage yield, drug release, and stability of lipid emulsions) were studied. The particle size of ATRA-loaded lipid emulsions was in the nano-size range of 124.4-378.2 nm with the narrow polydispersity index of 0.04-0.09, which decreased as the amount of co emulsifiers was increased. The amount of ATRA released from lipid emulsions was operated using a dialysis bag. The receptor medium was ethanol:polysorbate 80:water (10:15:75), adjusted to pH 8.5. ATRA release kinetics in this study were found to follow zero-order kinetics. As the concentration of co-emulsifiers increased, the flux of ATRA released from the lipid emulsions increased. In stability studies, the higher the amount of co-emulsifiers added, the lower the crystallization of ATRA was found. The percentage yield of ATRA was retained at about 70-90% and 60-72% after storage for 60 days at 4 degrees C and 25 degrees C, respectively. These results show a successful incorporation of ATRA into lipid emulsions with high loading capacity and good stability. PMID- 18410048 TI - Influence of beta-cyclodextrin complexation on glipizide release from hydroxypropyl methylcellulose matrix tablets. AB - Glipizide was complexed with beta-cyclodextrin in an attempt to enhance the drug solubility. The phase solubility diagram was classified as A(L) type, which was characterized by an apparent 1:1 stability constant that had a value of 413.82 M( 1). Fourier transform infrared spectrophotometry, differential scanning calorimetry, powder x-ray diffractometry and proton nuclear magnetic resonance spectral analysis indicated considerable interaction between the drug and beta cyclodextrin. A 2(3) factorial design was employed to prepare hydroxypropyl methylcellulose (HPMC) matrix tablets containing the drug or its complex. The effect of the total polymer loads (X1), levels of HPMC K100LV (X9), and complexation (X3) on release at first hour (Y1), 24 h (Y2), time taken for 50% release (Y3), and diffusion exponent (Y4) was systematically analyzed using the F test. Mathematical models containing only the significant terms (P < 0.05) were generated for each parameter by multiple linear regression analysis and analysis of variance. Complexation was found to exert a significant effect on Y1, Y2, and Y3, whereas total polymer loads significantly influenced all the responses. The models generated were validated by developing two new formulations with a combination of factors within the experimental domain. The experimental values of the response parameters were in close agreement with the predicted values, thereby proving-the validity of the generated mathematical models. PMID- 18410049 TI - Administration of i.v. chemotherapy & biotherapy agents. PMID- 18410050 TI - New faces, new beginnings. PMID- 18410051 TI - RWJF launches national program to build next generation of academic nurse leaders. PMID- 18410054 TI - Practical strategies for preventing adolescent suicide. PMID- 18410053 TI - Dental care for underserved children: what can be done? PMID- 18410055 TI - A glimpse at the holistic approach to wellness. PMID- 18410056 TI - Obesity wars continue. Home-based pediatric weight management. AB - There is a growing demand for pediatric weight management programs that can be adminisetred at home. School staff, including school nurses and food service staff, are frustrated by the pressure of handling these problems exclusively in the school; parents who want to exercise successful influence over their child's food choices are at a loss for answers. A pilot study of the SUCCESS AT HOME Program indicated that children made significant improvements in a number of health-related areas (weight, body-mass index, body circumference measures, flexibility) after nine months of using the program. Overall, parents were highly satisfied with the program and saw significant improvements in the physical health and psychological well-being of their children. PMID- 18410057 TI - Volunteering at diabetes camp: a professional experience with personal benefits. PMID- 18410058 TI - Healthy vision--what schools, parents and communities can do. PMID- 18410059 TI - [Beta-lactam antibiotics: when and how in acute otitis media and pharyngotonsillitis]. AB - Acute otitis media (AOM) and pharyngitis are very frequent diseases in pediatric parients. However, it is not always agreed which are the diagnostic criteria and empiric antibacterial treatment. It is often difficult to follow the different guidelines and not always can you choose between the different classes of antibiotics available. Furthermore, there's no unanimous agreement with respect to the most appropriate antibacterial agent to administer and the correct duration of the treatment. The aim of this article is to underline and discuss the emerging controversies in the empiric treatment of AOM and pharyngitis in pediatrics, with special attention to the use of beta-lactams in light of the most recent evidences regarding both clinical pharmacology and microbiology. PMID- 18410060 TI - [Childhood feeding, chronic-degenerative disease in adults, and nutrigenomics]. AB - Significant advances have been made in understanding the relation between dietary factors and disease prevention. However, the identification of those who will or will not benefit from dietary intervention strategies remains a major obstacle. The execution of the Human Genome Project has brought forth a wealth of information about the structure of the genome and the spectacular development of broad genomics technologies have catalyzed a new era in both medicine and nutrition. Each person is genetically unique and phenotypically distinct, and the genetic makeup that individuals inherit from their ancestors is responsible for variation in responses to food. Evidence continues to implicate dietary components and genetic susceptibilities as important determinants of chronic diseases, cancer risk and tumor behavior. Variation in incidence among and within populations with similar dietary patterns suggests that an individual's response may reflect interactions with genetic factors, which may modify gene, protein, and metabolite expression patterns. Nutrigenetics studies the genetic basis of the different individual responses to the same nutritional stimulus and Nutrigenomics is defined as the interaction between nutrition and an individual's genome. With the application of "omic" technologies, proteomic, metabolomic, transcriptomic, will increase our fundamental knowledge of the interaction between life processes and diet. The identification of diet-gene interactions will offers an opportunity to develop dietary interventions that will lead to evidence-based dietary strategies for restoring health and fitness, obviate the effects of genetic factors for preventing diet-related diseases and provide important clues about gene expression and gene modulation by environmental factors. PMID- 18410061 TI - [How to prepare a scientific paper]. PMID- 18410062 TI - [Epidemiology and clinical pictures of pharyngitis: report on the activity of a family paediatrician]. AB - OBJECTIVES: Pharyngitis (PM) is the most common child disease, that Family Paediatricians (FP) have to deal with. The aim of this study is to analyze epidemiology of PM comparing the validity of clinical scores and Rapid Antigen Detection Test (RADT) to diagnose Group A Streptococci PM. An evaluation of antibiotic prescription was performed too on the basis of the two groups. METHODS: A three-years epidemiological investigation was performed on a group of children observed in a FP office. Children was divided into two groups on the basis of the RADT (RADT+ and RADT-) and for each group statistical and epidemiological analysis was performed. RESULTS: A total of 1.445 children was gathered, 1.128 of them (78%) had RADT- and 317 (22%) RADT+. Data analysis shows significant statistical differences between data of children with RADT+, comparing to those of RADT-, regarding Milano Score and other clinical criteria, with the only exception of McIsaac Score. CONCLUSIONS: PM remains the most important children disease in an FP office. The use of RADT allows a rapids and correct etiological diagnosis and to contain to less than 50% antibiotic prescription. RADT is the diagnostic test to use in the PM practice, being easy and quick to perform, compared with clinical criteria and scores. PMID- 18410063 TI - Optimization of emergency department management of infants with bronchiolitis. AB - Bronchiolitis is the most common lower respiratory tract infection in infants < 2 years of age; in the last decades both incidence and hospitalization rate had increased, thus increasing sanitary burden. From November 2006 to March 2007, an experimental protocol was followed in the Emergency Department at G. Gaslini Children's Hospital, Genoa, Italy, which attempted to optimise the management of patients with bronchiolitis and to reduce the overall hospitalization rate therefore admitting only those patients with severe illness. All clinical evaluations of the patients were obtained administering a score (Bronchiolitis Clinical Score - BCS), to quantify both initial severity of illness and response to treatment. All patient were at first treated with inhaled epinephrine, supplemented with or substituted by other drugs, if needed, according to clinical evolution. Moreover, strict admission and discharge criteria were defined, taking into consideration the BCS, response to treatment and the presence of risk factors for severe disease, attempting to increase the role of the Short Stay Unit (SSU). The outcome evaluated were the percentage of patients discharged, admitted and managed through the SSU respectively, the length of stay and the readmission rate after discharge; data collected were then compared to that regarding patients with bronchiolitis presented at the ED from November 2005 to March 2006. Our data showed an increasing of both discharged patients (37.5% vs 25.22%) and patients managed through the SSU (25.83% vs 19.57%) and a related decrease of hospitalization (36.67% vs 55.22%); no significative difference was observed regarding the readmission rate between the two populations. We also observed a statistically significant reduction of the length of stay in the study population (2.07 +/- 2.56 vs 2.84 +/- 3.25, p = 0.005). In conclusion, the protocol proposed showed to be useful in optimizing the ED management of the patient with bronchiolitis, being able to safely reduce both admission rate and lenght of stay. PMID- 18410064 TI - [Hypocaloric diet and normocaloric diet in outpatient treatment in a group of children and adolescents]. AB - OBJECTIVE: The epidemic of obesity, mostly in pediatric age, is of increasing concern because of the impact of overweight on health status in adult life. We have evaluated the impact of two regimens (balanced hypocaloric diet or balanced normocaloric diet) in a group of children ad adolescents. MATERIAL AND METHODS: We have studied 260 patients seen at a community pediatric clinic. Of these, 45 overweight, 35 obese and 6 patients with normal weight were selected. The subjects were assigned to a hypocaloric balanced diet plus lifestyle changes or a normocaloric balanced diet plus lifestyle changes. MAIN RESULTS: In the obese patients on a hypocaloric diet, 64% of subjects remained in the "obese" category and 14% decided to shift to the "normocaloric diet". Also, the drop-out rate was 12%. In the obese patients on a normocaloric diet, 38% remained in the obese category, while 46% showed a reduction of their BMI to the "overweight" category. Moreover, the drop-out rate was lower (5%). CONCLUSION: These findings suggest that a more gentle approach to obesity/overweight in pediatric patients, with less restrictive nutritional interventions, could achieve a better compliance of the family, with a consistent reduction of overweight. PMID- 18410065 TI - [Early surgical therapy for idiopathic varicocele]. AB - The recurring questions about idiopathic varicocele are: a) timing of treatment; b) choice of therapeutical kind; c) influence of treatment on physiopathology of varicocele. In our experience, even if limited, the high ligation of spermatic vessels is simple, safe and effective, even at long term. Fifty-two boys under went surgical therapy at diagnosis--average age 12,9 y. We did not find recurrences of varicocele. Four boys showed postoperative hydrocele; no one required surgical treatment. The postoperative functional valuation, demonstrated a decreased quality of spermatograms in 9.1%. In conclusion, we suggest to early treatment of varicocele by means of a technique effective and devoid of complications. PMID- 18410066 TI - [Incontinentia pigmenti: case report]. AB - IP is an uncommon X-linked dominant disorder (incidence: 1/40.000 newborn). It is caused by mutations in NEMO. It is characterized by cutaneous lesions and dental, ocular, neurologic, nails, hair disorders. The ocular and neurologic sequelae represent the major morbidity in IP. We present a case-report with classical cutaneous features diagnostic for IP. The clinical, ophtalomologic and neurologic examinations revealed no other pathological manifestations. PMID- 18410067 TI - [Surgical reconstruction of skin layers of the leg in children: a case report]. PMID- 18410068 TI - Effect of the digenean parasites of fish on the fauna of Mediterranean lagoons. AB - Attention is drawn to the effects of parasites on their hosts, taking as a model the digenean parasites of teleosts (hereafter: fish) from lagoons along the French Mediterranean coast. Because digeneans have a heteroxenic life cycle, their impact is not limited to the definitive host, which harbours the sexual adults, but is extended to the first host (mollusc) and to the second host ("invertebrate" or fish). Adult parasites, in order to ensure efficient sexual reproduction, never cause excessive damage to their definitive host, usually only exploiting the intestinal fluids; however, the host must intensify its search for prey, which results in a diminished fitness. Within the first host, 'larval' stages of digenean parasites invade the gonads, resulting in its castration, then exhaustion and eventually death. The diversion of energy from the second hosts towards the parasites forces them to intensify their search for food, resulting in decreased fitness and an increased risk of being eaten; in addition, manipulation of the host's behaviour by parasites drives this host into the food chain of the definitive host. In lagoons, many individuals of almost all species of fish and invertebrates act as first, second and/or definitive hosts for digeneans. Obviously, parasites have a severe impact on the population dynamics of key taxa, on the food web and therefore also on the functioning of the whole lagoon ecosystem. Yet this impact has been largely overlooked or underestimated in functioning models, by ecologists, who tend to prioritize more apparent trophic relationships. PMID- 18410069 TI - Integrating genetic and parasitological approaches in the frame of multidisciplinary fish stock analysis. AB - To assess fish stocks boundaries and state, the tools of population genetics have been widely used, contributing to the evaluation of relevant parameters such as the identification of stock boundaries, the assessment of gene flow and the estimation of effective population size. Also, increasing evidences show that the monitoring of the genetic diversity level is a reliable method to check the status of fish stocks. However, genetics cannot answer all the questions. For example, in high gene flow species the genetic approach could have not enough resolution to identify stock limits, while the use of parasites as biological tags could provide insights into stock structure. Even better, the so-called holistic approach, applying simultaneously a wide range of complementary techniques, is the only one considered able to provide a reliable and complete picture of fish stocks and to address a sustainable exploitation of marine resources. The work will present some examples from multidisciplinary studies concerning commercially relevant species with different biological features: the demersal European hake (Merluccius merluccius), the small pelagic horse mackerel (Trachurus trachurus) and the large pelagic swordfish (Xiphias gladius). In all these case studies merging genetic, parasitological and environmental data helped to reveal the real patterns of stocks structure. PMID- 18410070 TI - Cophylogeny: insights from fish-parasite systems. AB - Host-parasite cophylogeny is a topic that has grasped the attention of scientists since the end of the 19th century, but the development of dedicated analytical methods only arose in the last 30 years. Research on host-parasite systems and on the development of more and more sophisticated numerical methods to estimate the degree of cospeciation has thus progressed, permitting the elaboration of evolutionary scenarios. The main outcome of these studies is that the expected clear pattern of cospeciation between many hosts and parasites is often obscure. In practice, much attention has been devoted to few host-parasite systems. Particularly, aquatic host-parasite associations have not been so extensively studied, and, after briefly reviewing the main analytical methods, this paper focuses on host-monogenean systems, because this kind of interaction is expected to be an ideal model for cophylogeny studies. But is it? And what does it tell us about the evolutionary and ecological forces driving cospeciation in the open sea? Biogeography should also be considered when possible, and it has been useful for explaining some patterns of cospeciation. It should thus be more deeply exploited in the future. We need new methods and new biological models that better, if not fully, depict patterns and thereby permit deeper understanding of processes within cophylogenetic patterns. PMID- 18410071 TI - Diversity and distribution of fish tapeworms of the "Bothriocephalidea" (Eucestoda). AB - Recent studies have demonstrated the invalidity of the Pseudophyllidea, a long term recognised order of tapeworms (Platyhelminthes: Cestoda), typical in possessing two dorsoventrally situated attachment organs called bothria. In fact, cestodes parasitic in tetrapods, especially mammals including man, form a relatively basal group called provisionally the "Diphyllobothriidea", whereas tapeworms occurring in freshwater and marine fish, with a few taxa known from amphibians (frogs and newts), belong to a more derived clade, for which the name "Bothriocephalidea" is tentatively proposed. Revision of the "Bothriocephalidea", based on literary data, study of type- and voucher specimens and extensive newly collected material made it possible to critically review the species composition of the group and to prepare a tentative list of its valid species. Out of 305 nominal taxa, only 125 species are considered to be valid. In addition, the spectrum of definitive hosts and geographical distribution of bothriocephalideans are briefly discussed. PMID- 18410072 TI - Molecular detection of sibling species in anisakid nematodes. AB - The number of sibling species of anisakid nematodes detected over the last two decades has been increased, fuelled by the use of genetic/molecular methodologies. In the present review, we summarize the biological species discovered within most of the nominal species belonging to the genera Anisakis, Contracaecum and Pseudoterranova by the use of allozyme (20-24 loci studied) and recently confirmed by us using mitochondrial cox-2 gene sequence analysis (mtDNA cox-2). Ecological evidence relating to the distributional range of the genetically detected sibling species and their host preferences, which represent data sets that can be utilized for species delimitation and definition, are summarized. PMID- 18410074 TI - Effects of a hurricane on fish parasites. AB - Hurricanes, also called tropical cyclones, can dramatically affect life along their paths, including a temporary losing or reducing in number of parasites of fishes. Hurricane Katrina in the northern Gulf of Mexico in August 2005 provides many examples involving humans and both terrestrial and aquatic animals and plants. Fishes do not provide much of an indicator of hurricane activity because most species quickly repopulate the area. Fish parasites, however, serve as a good indicator of the overall biodiversity and environmental health. The reasons for the noted absence or reduction of parasites in fishes are many, and specific parasites provide indications of different processes. The powerful winds can produce perturbations of the sediments harboring intermediate hosts. The surge of high salinity water can kill or otherwise affect low salinity intermediate hosts or free-living stages. Both can introduce toxicants into the habitat and also interfere with the timing and processes involved with host-parasite interrelationships. All these have had a major influence on fish parasite populations of fishes in coastal Mississippi, especially for those parasites incorporating intermediate hosts in their life cycles. The length of time for a parasite to become re-established can vary considerably, depending on its life cycle as well as the associated biota, habitat, and environmental conditions, and each parasite provides a special indicator of environmental health. PMID- 18410073 TI - Nematode parasites of fishes: recent advances and problems of their research. AB - Although nematodes (Nematoda) belong to the most frequent and the most important parasites of fishes in the freshwater, brackish-water and marine environments throughout the world, the present knowledge of these parasites remains still incomplete, especially as to their biology and ecology, but also taxonomy, phylogeny, zoogeography, and the like. However, a certain progress in the research of fish nematodes has been achieved during recent years. An overview of some of the most important discoveries and results obtained is presented. As an example, existing problems in the taxonomy of these nematodes are shown in the dracunculoid family Philometridae (presently including 109 species in 9 genera), where they are associated mainly with some biological peculiarities of these mostly important tissue parasites. Nematodes of the Dracunculoidea as a whole remain poorly known; for example, of 139 valid species parasitizing fishes, 81 (58%) are known by females only and the males have not yet been described for members of 8 (27%) of genera. A taxonomic revision of this nematode group, based on detailed morphological, life history and molecular studies of individual species, is quite necessary; for the time being, Moravec (2006) has proposed a new classification system of dracunculoids, where, based on previous molecular studies, the Anguillicolidae is no longer listed in Dracunculoidea, but in an independent superfamily Anguillicoloidea. Important results have recently been achieved also in the taxonomy of fish nematodes belonging to other superfamilies, as well as in studies of their geographical distribution and diversity in different parts of the world and those of their biology. Opportunities for more detailed studies of fish nematodes have recently greatly improved with the use of some new methods, in particular SEM and DNA studies. There is a need to create a new classification system of these parasites reflecting phylogenetic relationships; a prerequisite for this is taxonomic revisions of different groups based on detailed studies of individual species, including mainly their morphology, biology and genetics. Further progress should concern studies on various aspects of biology, ecology and host-parasite relationships, because these data may have practical implications. PMID- 18410075 TI - The structure of parasite communities in fish hosts: ecology meets geography and climate. AB - Parasite communities in fish hosts are not uniform in space: their diversity, composition and abundance vary across the geographical range of a host species. Increasingly urgently, we need to understand the geographic component of parasite communities to better predict how they will respond to global climate change. Patterns of geographical variation in the abundance of parasite populations, and in the diversity and composition of parasite communities, are explored here, and the ways in which they may be affected by climate change are discussed. The time has come to transform fish parasite ecology from a mostly descriptive discipline into a predictive science, capable of integrating complex ecological data to generate forecasts about the future state of host-parasite systems. PMID- 18410076 TI - Host-parasite interactions from an ecotoxicological perspective. AB - In recent years there has been an increasing number of papers showing how parasitism and pollution can interact with each other in aquatic organisms. Apart from parasitological aspects these interactions are also important in terms of ecotoxicological research. The current presentation aims at identifying three promising directions for future research in the interdisciplinary field of parasitology and ecotoxicology. 1. Parasites as sinks for pollutants within their hosts: Some parasites are able to reduce pollutant levels in the tissues of their host. The reduction of pollutants is an interesting implication since parasites are beneficial to their hosts from this perspective. In other cases free-living accumulation indicators may erroneously indicate low levels of pollution if they are infected with parasites. 2. Parasites as a diagnostic tool to test bioavailability of substances. In order to take up and accumulate pollutants the substances have to be metabolized by the host first. Accordingly, the detection of substances within endoparasites is a sign for the biological availability of pollutants. 3. Changes of biomarker responses of the host against pollutants. Parasites can alter physiological reactions of their hosts against pollutants in different ways. Therefore, in ecotoxicological studies, examining the question whether exposure to certain chemicals affects the physiological homeostasis of a test organism, it is important to use organisms that are known to be uninfected. PMID- 18410077 TI - Helminths and protozoans of aquatic organisms as bioindicators of chemical pollution. AB - There is no doubt that the aquatic environments receive large quantities of chemicals as consequence of human activities and that those substances have a detrimental effect on human health. Despite the obvious need for effective disposal of these substances, we need to understand and prevent the outcome of harmful environmental exposures. Thus, we need biomarkers and bioindicators to advance our understanding to these harmful exposures and their biological effects. In the last three decades a large number of publications has suggested that aquatic organisms and their parasites (mainly helminths and ciliate protozoans) are useful bioindicators of chemical pollution. However, the main weakness of this approach is that after exposure the population size of these parasites can increase or decrease without a consistent pattern. I suggest that this is in part due to the lack of focus on the correct spatial or temporal scales at which the environment is acting over our study object. Thus, I propose to use spatially explicit (= georeferenced) data for determining whether there is spatial structure in our study area. Spatial structure is the tendency of nearby samples to have attribute values more similar than those farther apart. These attributes are shaped by environmental variables acting at specific spatial and temporal scales. Thus, I suggest to consider these tools for determining the correct spatial or temporal scales of study, but also to record pollutant concentrations, bioindicators, biomarkers and parasites at individual host level. Combining this information with long-term monitoring programs is likely to improve our understanding of the effects of chemical pollutants over the aquatic environments. PMID- 18410078 TI - Protective immunity in fish against protozoan diseases. AB - The demand for and costs of producing land-based animal protein continues to escalate as the world population increases. Fish is an excellent protein, but the catch-fishery is stagnant or in decline. Intensive cage culture of fish is a viable option especially in countries with lakes/rivers and/or a long coastline; however, disease outbreaks will likely occur more frequently with cage culture. Hence protective strategies are needed, and one approach is to exploit the piscine immune system. This discussion highlights immunity (innate/natural and adaptive/acquired) in fish against three pathogenic protozoa (Amyloodinium ocellatum, Ichthyophthirius multifiliis and Cryptobia salmositica). Histone-like proteins in the mucus and skin of naturally resistant fish kill trophonts of A. ocellatum, and also may cause abnormal development of tomonts. Breeding of Cryptobia-resistant brook charrs is possible as resistance is controlled by a dominant Mendelian locus, and the parasite is lysed via the Alternative Pathway of Complement Activation. Production of transgenic Cryptobia-tolerant salmon is an option. Recovered fish are protected from the three diseases (acquired immunity). Live I. multifiliis theronts injected intraperitoneally into fish elicit protection. Also, a recombinant immoblizing-antigen vaccine against ichthyophthirosis has been developed but further evaluations are necessary. The live Cryptobia vaccine protects salmonids from infections while the DNA-vaccine stimulates production of antibodies to neutralize the disease causing factor (metalloprotease) in cryptobiosis; hence infected fish recover more rapidly. PMID- 18410079 TI - Parasitic diseases of cultured marine finfishes and their surveillance in China. AB - More than 40 species of marine fishes are cultured in China and a wide variety of parasites are reported as lethal pathogens of these fishes in culture conditions. In the case of net cages, the culture facilities provides a good substrate for monogenean eggs to become entangled and the intensive aggregation of fishes facilitates the transmission of parasites between hosts. Relatively thorough studies on parasitic pathogens of marine fishes in China predominately concern the ciliate Cryptocaryon irritans and capsalid monogeneans (mainly Benedenia sp. and Neobenedenia sp.). Although nearly all such reports are related to treatment procedures, no single method has proved to be adequate for the effective control of these parasitic pathogens in marine cultured fishes. The National Fisheries Technology Extension Center (NFTEC) has established surveillance systems to monitor the diseases of aquaculture, including the parasitic diseases of maricultured fishes. The national monitoring stations for diseases of cultured marine fishes are distributed in the coastal counties or cities and provide remote in situ diagnoses of diseased fishes. International cooperation and effort are required for the control of parasitic diseases of marine finfish because of both the increasing international trade of eggs (seed) and larvae and commercial products in terms of live marine finfishes, which can readily result in the transmission of pathogens. PMID- 18410080 TI - Predicting binding modes from free energy calculations. AB - To produce reliable predictions of bioactive conformations is a major challenge in the field of structure-based inhibitor design and is a requirement for accurate binding free energy predictions with structure-based methods. A series of HIV-1 reverse transcriptase inhibitors was cross-docked using a non-native crystal structure that resulted in two distinct clusters of possible conformations. One of these clusters was compatible with an existing crystal structure, whereas the other displayed a flipped heterocyclic group. Binding free energies, using the non-native crystal structure, calculated from several scoring functions, were similar for the two clusters, and no conclusion about the binding mode could be drawn from these results. The two clusters could be separated through rescoring with the linear interaction method (LIE) in combination with molecular dynamics simulations, which leads to a binding mode prediction in line with experimental crystallographic data. Further, the LIE model produces the best correlation between experimental and calculated binding free energies among the tested scoring methods. PMID- 18410081 TI - Identification of Plasmodium falciparum spermidine synthase active site binders through structure-based virtual screening. AB - Seven novel binders, binding in the active site of Plasmodium falciparum spermidine synthase, were identified by structure-based virtual screening. The binding of these compounds was experimentally verified by NMR techniques. Spermidine synthase, an enzyme involved in the polyamine pathway, has been suggested as a target for treating malaria. The virtual screening protocol combined 3D pharmacophore filtering, docking, and scoring, focusing on finding compounds predicted to form interactions mimicking those of a previously known binder. The virtual screen resulted in the selection of 28 compounds that were acquired and tested from 2.6 million starting structures. Two of the seven binders were predicted to bind in the amino substrate binding pocket. Both of these showed stronger binding upon addition of methylthioadenosine, one of the two products of the enzyme, and a known binder and inhibitor. The five other compounds were predicted to bind in the part of the active site where the other substrate, decarboxylated S-adenosylmethionine, binds. These five compounds all competed for binding with methylthioadenosine. PMID- 18410082 TI - Bioisosteric heterocyclic versions of 7-{[2-(4-phenyl-piperazin-1 yl)ethyl]propylamino}-5,6,7,8-tetrahydronaphthalen-2-ol: identification of highly potent and selective agonists for dopamine D3 receptor with potent in vivo activity. AB - In the current report, we extend the SAR study on our hybrid structure 7-{[2-(4 phenyl-piperazin-1-yl)ethyl]propylamino}-5,6,7,8-tetrahydronaphthalen-2-ol further to include heterocyclic bioisosteric analogues. Binding assays were carried out with HEK-293 cells expressing either D2 or D3 receptors with tritiated spiperone to evaluate inhibition constants (Ki). Functional activity of selected compounds in stimulating GTPgammaS binding was assessed with CHO cells expressing human D2 receptors and AtT-20 cells expressing human D3 receptors. The highest binding affinity and selectivity for D3 receptors were exhibited by (-) 34 (Ki=0.92 nM and D2/D3=253). In the functional GTPgammaS binding assay, (-)-34 exhibited full agonist activity with picomolar affinity for D3 receptor with high selectivity (EC50=0.08 nM and D2/D3=248). In the in vivo rotational study, (-)-34 exhibited potent rotational activity in 6-OH-DA unilaterally lesioned rats with long duration of action, which indicates its potential application in neuroprotective treatment of Parkinson's disease. PMID- 18410083 TI - Ring size in octreotide amide modulates differently agonist versus antagonist binding affinity and selectivity. AB - H-DPhe (2)-c[Cys (3)-Phe (7)-DTrp (8)-Lys (9)-Thr (10)-Cys (14)]-Thr (15)-NH2 (1) (a somatostatin agonist, SRIF numbering) and H-Cpa (2)-c[DCys (3)-Tyr (7)-DTrp (8)-Lys (9)-Thr (10)-Cys (14)]-Nal (15)-NH2 (4) (a somatostatin antagonist) are based on the structure of octreotide that binds to three somatostatin receptor subtypes (sst 2/3/5) with significant binding affinity. Analogues of 1 and 4 were synthesized with norcysteine (Ncy), homocysteine (Hcy), or D-homocysteine (DHcy) at positions 3 and/or 14. Introducing Ncy at positions 3 and 14 constrained the backbone flexibility, resulting in loss of binding affinity at all sst s. The introduction of Hcy at positions 3 and 14 improved selectivity for sst 2 as a result of significant loss of binding affinity at the other sst s. Substitution by DHcy at position 3 in the antagonist scaffold (5), on the other hand, resulted in a significant loss of binding affinity at sst 2 and sst 3 as compared to the different affinities of the parent compound (4). The 3D NMR structures of the analogues in dimethylsulfoxide are consistent with the observed binding affinities. PMID- 18410086 TI - Nanographenes as active components of single-molecule electronics and how a scanning tunneling microscope puts them to work. AB - Single-molecule electronics, that is, realizing novel electronic functionalities from single (or very few) molecules, holds promise for application in various technologies, including signal processing and sensing. Nanographenes, which are extended polycyclic aromatic hydrocarbons (PAHs), are highly attractive subjects for studies of single-molecule electronics because the electronic properties of their flat conjugated systems can be varied dramatically through synthetic modification of their sizes and topologies. Single nanographenes provide high tunneling currents when adsorbed flat onto conducting substrates, such as graphite. Because of their chemical inertness, nanographenes interact only weakly with these substrates, thereby preventing the need for special epitaxial structure matching. Instead, self-assembly at the interface between a conducting solid, such as the basal plane of graphite, and a nanographene solution generally leads to highly ordered monolayers. Scanning tunneling spectroscopy (STS) allows the current-voltage characteristics to be measured through a single molecule positioned between two electrodes; the key to the success of STS is the ability to position the scanning tunneling microscopy (STM) tip freely with respect to the molecule in all dimensions, that is, both parallel and perpendicular to the surface. In this Account, we report the properties of nanographenes having sizes ranging from 0.7 to 3.1 nm and exhibiting various symmetry, periphery, and substitution types. The size of the aromatic system and the nature of its perimeter are two essential features affecting its HOMO-LUMO gap and charge carrier mobility in the condensed phase. Moreover, the extended pi area of larger substituted PAHs improves the degree of self-ordering, another key requirement for high-performance electronic devices. Self-assembly at the interface between an organic solution and the basal plane of graphite allows deposition of single molecules within the well-defined environment of a molecular monolayer. We have used STM and STS to investigate both the structures and electronic properties of these single molecules in situ. Indeed, we have observed key electronic functions, rectification and current control through single molecules, within a prototypical chemical field-effect transistor at ambient temperature. The combination of nanographenes and STM/STS, with the PAHs self-assembled in oriented molecular mono- or bilayers at the interface between an organic solution and the basal plane of graphite and contacted by the STM tip, is a simple, reliable, and versatile system for developing the fundamental concepts of molecular electronics. Our future targets include fast reversible molecular switches and complex molecular electronic devices coupled together from several single-molecule systems. PMID- 18410085 TI - Combining docking, molecular dynamics and the linear interaction energy method to predict binding modes and affinities for non-nucleoside inhibitors to HIV-1 reverse transcriptase. AB - Docking, scoring, molecular dynamics (MD), and the linear interaction energy (LIE) method are used here to predict binding modes and affinities for a set of 43 non-nucleoside inhibitors to HIV-1 reverse transcriptase. Starting from a crystallographic structure, the binding modes of 43 inhibitors are predicted using automated docking. The Goldscore scoring function and the LIE method are then used to determine the relative binding free energies for the inhibitors. The Goldscore scoring function does not reproduce the relative binding affinities for the inhibitors, while the standard parametrization of the LIE method reproduces the experimental binding free energies for 39 inhibitors with an R (2) = 0.70 and an unsigned average error of 0.8 kcal/mol. The present calculations provide a validation of the combination of docking, MD, and LIE as a powerful tool in structure-based drug design, and the methodology is easily scalable for attaining a higher throughput of compounds. PMID- 18410084 TI - Ring size of somatostatin analogues (ODT-8) modulates receptor selectivity and binding affinity. AB - The synthesis, biological testing, and NMR studies of several analogues of H c[Cys (3)-Phe (6)-Phe (7)-DTrp (8)-Lys (9)-Thr (10)-Phe (11)-Cys (14)]-OH (ODT-8, a pan-somatostatin analogue, 1) have been performed to assess the effect of changing the stereochemistry and the number of atoms in the disulfide bridge on binding affinity. Cysteine at positions 3 and/or 14 (somatostatin numbering) were/was substituted with d-cysteine, norcysteine, D-norcysteine, homocysteine, and/or D-homocysteine. The 3D structure analysis of selected partially selective, bioactive analogues (3, 18, 19, and 21) was carried out in dimethylsulfoxide. Interestingly and not unexpectedly, the 3D structures of these analogues comprised the pharmacophore for which the analogues had the highest binding affinities (i.e., sst 4 in all cases). PMID- 18410087 TI - New synthetic routes to a disulfidodinickel(II) complex: characterization and reactivity of a Ni2(mu-eta2:eta2-S2) core. AB - Activation of elemental sulfur by the monovalent nickel complex [PhTt (tBu)]Ni(CO) [PhTt(tBu)=phenyl{tris[(tert-butylmethyl)thio]methyl}borate] generates the disulfidodinickel(II) complex 2. This species is alternatively accessible via thermal decomposition of [PhTt (tBu)]Ni(SCPh3). Spectroscopic, magnetic, and X-ray diffraction studies establish that 2 contains a mu eta(2):eta(2)-S2 ligand that fosters antiferromagnetic exchange coupling between the Ni (II) ions. This observation is in contrast to the lighter congener, oxygen, which strongly favors the bis(mu-oxo)dinickel(III) structure. 2 oxidizes PPh 3 to SPPh3 and reacts with O2, generating several products, one of which has been identified as [(PhTt (tBu))Ni]2(mu-S) (3). PMID- 18410088 TI - Synthesis of amidolanthanides with new chiral biaryl-based NNO ligands and their use as catalysts for enantioselective hydroamination/cyclization. AB - A new series of amidolanthanides have been prepared from the reactions between Ln[N(SiMe3)2]3 and the chiral NNO ligands, (S)-2-(pyrrol-2-ylmethyleneamino)-2' hydroxy-6,6'-dimethyl-1,1'-biphenyl (2H2) and (S)-5,5',6,6',7,7',8,8'-octahydro-2 (pyrrol-2-ylmethyleneamino)-2'-hydroxy-1,1'-binaphthyl (3H2), which are synthesized from the condensation of pyrrole-2-carboxaldehyde with 1 equiv of (S) 2-amino-2'-hydroxy-6,6'-dimethyl-1,1'-biphenyl or (S)-5,5',6,6',7,7',8,8' octahydro-2-amino-2'-hydroxy-1,1'-binaphthyl, in the presence of molecular sieves at 70 degrees C, respectively. Treatment of 2H2 with 1 equiv of Ln[N(SiMe3)2]3 (Ln=Sm, Yb) in toluene under reflux, followed by recrystallization from a toluene solution, gives the dimeric amido complexes, {2-SmN(SiMe3)2}2.0.5C7H8 (6.0.5C7H8) and {2-YbN(SiMe3)2} 2.1.5C7H8(8.1.5C7H8), in good yields. While under similar reaction conditions, the reaction of 2H2 with 1 equiv of Y[N(SiMe3)2]3 leads to the isolation of a mixture of {2-YN(SiMe3)2}2 (7a) and {(2)2Y}Y[N(SiMe3)2]2(7b) in 82% total yield; the reaction of 3H2 with 1 equiv of Ln[N(SiMe3)2]3 (Ln=Y, Yb) gives the trinuclear complexes, {(3)2Ln}2LnN(SiMe3) 2.1.5C7H8 (Ln=Y(9.1.5C7H8), Yb(10.1.5C7H8)), in good yields. All compounds have been characterized by various spectroscopic techniques and elemental analyses. The solid-state structures of compounds 2H2 and 6- 10 have been further confirmed by X-ray diffraction analyses. Complexes 6- 9 are active catalysts for the asymmetric hydroamination/cyclization of aminoalkenes, affording cyclic amines in good yields with moderate ee values. PMID- 18410089 TI - From large 12-membered macrometallacycles to ionic (NHC)2M+Cl- type complexes of gold and silver by modulation of the N-substituent of amido-functionalized N heterocyclic carbene (NHC) ligands. AB - A series of structurally diverse gold and silver complexes extending from ionic (NHC) 2M(+)Cl(-) (M=Au, Ag) type complexes to large 12-membered macrometallacycles have been prepared by the appropriate modification of the N substituent of amido-functionalized N-heterocyclic carbenes. Specifically, the ionic, [1-(R)-3-{ N-(t-butylacetamido)imidazol-2-ylidene}]2M(+)Cl(-), (R=t-Bu, i Pr; M=Au, Ag; 1b, 1c, 2b, 2c) complexes, were obtained in case of the N- t-butyl substituent of the amido-functionalized sidearm while 12-membered macrometallacycles, [1-(R)-3-{N-(2,6-di i-propylphenylacetamido)imidazol-2 ylidene}]2M2, (R=t-Bu, i-Pr; M=Au, Ag; 3b, 3c, 4b, 4c) were obtained in case of the 2,6-di i-propylphenyl N-substituent. These structurally diverse complexes of gold and silver were, however, prepared employing a common synthetic pathway involving the reactions of the imidazolium chloride salts (1a, 2a, 3a, 4a) with Ag2O to give the silver complexes (1b, 2b, 3b, 4b) and which, when treated with (SMe2)AuCl, gave the gold complexes (1c, 2c, 3c, 4c). Detailed density functional theory studies of 1b, 1c, 2b, 2c, 3b, 3c, 4b, and 4c were carried out to gain insight about the structure, bonding, and the electronic properties of these complexes. The NHC-metal interaction in the ionic 1b, 1c, 2b, and 2c complexes is primarily composed of the interaction of the carbene lone pair with the empty p orbital of the metal (5p for Ag and 6p for Au) while the same in the macrometallacyclic 3b, 3c, 4b, and 4c complexes consisted of the interaction of the carbene lone pair with the empty s orbital of the metal (5s for Ag and 6s for Au). The observation of a low energy emission in about the 580-650 nm region has been tentatively assigned to originate from the presence of weak metallophilic interaction in these macrometallacyclic 3b, 3c, 4b, and 4c complexes. PMID- 18410090 TI - Supramolecular adsorption of alkaloids by metallosalphen complexes. AB - Mono and bis-zinc(II)-centered salphen derivatives 1-5 are presented as efficient adsorption materials for pyridine-based alkaloid derivatives. The different alkaloid assemblies were studied by UV-vis and NMR spectroscopy, and high binding constants (Ks approximately 10(5)) were additionally determined for the supramolecular complexes based on nicotine. X-ray analyses furthermore revealed, together with spectroscopic solution data, a preferential positioning of the nicotine guest(s). Upon binding to bis-Zn(II)-bis-salphen complexes, the dinicotine assembly provokes a colorimetric change that may be useful for colorimetric analyses. The adsorption/desorption process of nicotine was studied using a polymeric bis-Zn(salphen) complex (5) and showed a recycling potential of this type of complexes in the binding of alkaloid compounds. PMID- 18410091 TI - Single-molecule magnets: a family of MnIII/CeIV complexes with a [Mn8CeO8]12+ core. AB - Four heterometallic, enneanuclear Mn8Ce clusters [Mn8CeO8(O2CMe)12(H2O)4] (4), [Mn8CeO8(O2CMe)12(py)4] (5), [Mn8CeO8(O2CPh)12(MeCN)4] [Mn8CeO8(O2CPh)12(dioxane)4] (6), and [Mn8CeO8(O2CCHPh2)12(H2O)4] (7) have been prepared by various methods. Their cores are essentially isostructural and comprise a nonplanar, saddlelike [MnIII8O8]8+ loop containing a central CeIV ion attached to the eight micro3-O2- ions. Peripheral ligation around the [Mn8CeO8]12+ core is provided by eight micro- and four micro3-O2CR- groups. Terminal ligation on four MnIII atoms is provided by H2O in 4 and 7, pyridine in 5, and MeCN/dioxane in 6. Solid-state magnetic susceptibility studies, fits of dc magnetization vs field and temperature data, and in-phase ac susceptibility studies in a zero dc field have established that complexes 4, 5, and 7 possess S=16, S=4 or 5, and S=6+/-1 spin ground states, respectively, but in all cases there are very low-lying excited states. The large variation in the ground-state spins for this isostructural family is rationalized as due to a combination of weak exchange interactions between the constituent MnIII atoms, and the presence of both nearest-neighbor and next-nearest-interactions of comparable magnitudes. Magnetization vs applied dc field sweeps on single crystals of 4.4H2O and 7.4H2O.3MeCN.2CH2Cl2 down to 0.04 K have established that these two complexes are new single-molecule magnets (SMMs). The former also shows an exchange-bias, a perturbation of its single-molecule properties from very weak intermolecular interactions mediated by hydrogen-bonding interactions with lattice-water molecules of crystallization. PMID- 18410092 TI - Modulating affinities of di-2-picolylamine (DPA)-substituted quinoline sensors for zinc ions by varying pendant ligands. AB - We have developed a series of di-2-picolylamine (DPA)-substituted quinoline sensors, HQ1- 4, bearing a pendant ligand at the 8 position of quinoline. UV-vis spectra of HQ1- 4 showed similar variations to that of HQ5 but with different varying extents upon the titration of zinc ions. Fluorescence intensities of HQ1, HQ3, and HQ4 were enhanced 4-6 times upon the addition of 1 equiv of zinc ions under an aqueous buffer. Somewhat unexpectedly, HQ2 is nonfluorescent in the presence of metal ions, including zinc ions. The affinities of HQ sensors are distributed in a broad range from nanomolarity to femtomolarity by varying the pendant ligands near the coordination unit. More importantly, these new sensors exhibited very high selectivity for Zn(2+) over Na(+), K(+), Mg(2+), and Ca(2+) at the millimolar level and over other transition metal ions at the micromolar level, except for Cd(2+). These findings indicated that the incorporations of the pendant groups exerted no effect on the spectroscopic properties and selectivity of the parent fluorescent sensor, with the exception of HQ2. Finally, X-ray crystal structures of ZnHQ's revealed that the auxiliary pendant groups at the 8 position participated in zinc coordination and were able to tune the affinities of HQ sensors. PMID- 18410093 TI - Charge, orbital, and magnetic ordering in YBaFe2O5 from first-principles calculations. AB - First principles calculations using the augmented plane wave plus local orbitals method, as implemented in the WIEN2k code, have been used to investigate the electronic and magnetic properties of YBaFe2O5, especially as regards the charge orbital ordering. Although the total 3d charge disproportion is rather small, an orbital order parameter defined as the difference between t2g orbital occupations of Fe2+ and Fe3+ cations is large (0.73) and gives unambiguous evidence for charge and orbital ordering. Strong hybridization between O2p and Fe e g states results in the nearly complete loss of the separation between the total charges at the Fe2+ and Fe3+ atoms. Furthermore, the relationship between the orbital ordering and charge ordering is also discussed. The dxz orbital ordering is responsible for the stability of the G-type antiferromagnetic spin ordering and the charge ordering pattern. PMID- 18410094 TI - Self-assembly of luminescent Sn(IV)/Cu/S clusters using metal thiolates as metalloligands. AB - Through the use of (Bu4N)2[Sn3S4(edt)3] (edt=SCH2CH2S(2-)) and Sn(SPh)4 as metalloligands, three neutral compounds have been obtained: [(Ph3P) 2Cu] 2SnS(edt)(2).2CH2Cl2.H2O (1a), [(Ph3P) 2Cu]2SnS(edt)2.2DMF.H2O (1b), and [(Ph3P)Cu] 2Sn(SPh)(6).3H 2O (2). Single-crystal X-ray diffraction studies revealed that compounds 1a and 1b contain the same neutral butterfly-like [(Ph3P)2Cu]2SnS(edt)2 cluster, which consists of one central SnS 5 dreich trigonal bipyramid sharing one vertex and two sides with two slightly distorted CuS 2P2 tetrahedrons. Compound 2 has a linear [(Ph3P)Cu]2Sn(SPh)6 cluster that is composed of a central distorted SnS 6 octahedron sharing two opposite planes with two slightly distorted CuS 3P tetrahedrons. Compound 1a exhibited an emission at 568 nm (tau=12.86 micros) in the solid state, while in CH 2Cl 2 solution, 1a exhibited a green emission at 534 nm (tau=4.75 micros). Compound 2 showed an intense red emission at 696 nm (tau=3.64 micros) upon excitation at 307 nm in the solid state. PMID- 18410095 TI - Copper-complex-linked polytungsto-bismuthate (-antimonite) chain containing sandwich Cu(II) ions partially modified with imidazole ligand. AB - Two sandwich-type complexes Na9n(Cu(im)4(H2O)2)1.5n(Cu(im)4(H2O))n[{Cu(im)4}{Na(H2O)2}3{Cu3(im)2(H2O)}(XW9O33 2]2n .(xH2O)n (im=imidazole, X=Bi (1), Sb(2), x=42.5 (1), 40 (2)) have been synthesized and structurally characterized. Basic frameworks of 1 and 2 are built from sandwich-type [{Na(H2O) 2} 3{Cu3(im)2(H2O)}(XW9O33)2](9-) (X=Bi or Sb) anions and [Cu(im) 4](2+) cations. The Cu(2+) and Na(+) ions in the central belt are coordinated by alpha-[XW9O33](9-) units, im, and water molecules to form {CuO4(im)}, {CuO4(H2O)}, and {NaO4(H2O)2} groups in which Cu (2+) ions are partially modified with im ligands. These groups connect alternately forming a six-membered ring including six alpha-[XW9O 33](9-) units. Neighboring anions are further linked by [Cu(im) 4](2+) cations to display an unprecedented anionic chain, which is first observed in sandwich-type tungsto-bismuthate (-antimonite) system. Two kinds of isolated copper complexes and sodium ions are located as counterions, which cause three-dimensional packings of 1 and 2 to present interesting cage structures. The magnetic properties for 1 and 2 both indicate dominant antiferromagnetic interactions among trinuclear Cu(II) clusters. PMID- 18410096 TI - A diastereo- and enantioselective synthesis of alpha-substituted syn-alpha,beta diamino acids. AB - Highly diastereo- and enantioselective additions of substituted alpha-nitroesters to imines have been developed. High diastereoselection relies on the finding that the combination of chiral proton catalyst 2b and alpha-nitro aryl esters bearing 2,6-disubstitution combine to raise substrate-controlled diastereoselection to >20:1 in favor of the syn diastereomer. Furthermore, the chiral catalyst provides enantioselection to the 99% level through control of the addition step in which the azomethine pi-faces are differentiated. The bifunctional chiral protic acid catalyst enables these reactions to proceed without separate preactivation of either substrate, leading to a straightforward synthetic protocol for the formation of alpha,beta-diamino phenyl alanine derivatives. PMID- 18410097 TI - Capturing transient structures in the elimination reaction of haloalkane in solution by transient X-ray diffraction. AB - This Communication reports simultaneous tracking of structural and kinetic information for the photoinduced elimination reaction of 1,2 diiodotetrafluoroethane in solution by transient X-ray diffraction. The transient structure of .CF2CF2I is determined to be a classical mixture whereas .CH2CH2I is bridged. Compared with the gas phase reaction, the secondary dissociation of .CF2CF2I into C2F4 and I is slowed down by a factor of 6 in solution. Transient X ray diffraction offers a complementary method for capturing transient structures in solution which might be invisible or "optically silent" in time-resolved optical spectroscopy. PMID- 18410098 TI - Oxygen-atom transfer from carbon dioxide to a Fischer carbene at (PNP)Ir. AB - Dehydrogenation of the dihydride (PNP)IrH2 with norbornylene in the presence of t butyl methyl ether leads to formation of an iridium(I) Fischer carbene complex, (PNP)Ir C(H)OtBu, by double C-H activation and loss of H2. The square planar pincer-type carbene effects quantitative oxygen-atom transfer from CO2 (1 atm) at ambient temperature to generate t-butyl formate and (PNP)Ir-CO. The iridium carbene reacts similarly with carbonyl sulfide and phenyl isocyanate, causing sulfur-atom and nitrene-group transfer, respectively. In the absence of a hydrogen acceptor, thermolysis of (PNP)IrH2 in t-butyl methyl ether under an atmosphere of CO2 also results in the formation of (PNP)Ir-CO and oxidation of t butyl methyl ether to t-butyl formate via an iridium carbene. Preliminary mechanistic studies indicate that these reactions proceed through an intermediate four-membered metallalactone. PMID- 18410099 TI - Iron-catalyzed direct arylation through directed C-H bond activation. AB - An iron-catalyzed C-C bond formation reaction of a nitrogen-containing aromatic compound with an arylzinc reagent takes place at 0 degrees C in a good to quantitative yield. The reaction involves a C-H bond activation directed by a neighboring nitrogen atom. The important additives in this reaction are 1,10 phenanthroline, tetramethylethylenediamine, and 1,2-dichloro-2-methylpropane, in the absence of which a very low product yield was observed. PMID- 18410100 TI - Synthesis and structure verification of the vaccine adjuvant QS-7-Api. Synthetic access to homogeneous Quillaja saponaria immunostimulants. AB - QS-7-Api is an exceedingly potent immuno-adjuvant isolated from the bark of Quillaja saponaria. It is significantly less toxic than QS-21, a related saponin that is currently the favored adjuvant in anticancer and antiviral vaccine clinical trials. Tedious isolation/purification protocols and uncertainty in its structural constitution have hindered the clinical development of QS-7. A chemical synthesis of QS-7-Api is described, providing structural verification of the adjuvant. A novel semisynthetic sequence to QS-7-Api has also been established, greatly facilitating access to QS-7 for preclinical and clinical evaluation. PMID- 18410101 TI - Tertiary aromatic amide for memory of chirality: access to enantioenriched alpha substituted valine. AB - A new methodology for the asymmetric synthesis of quaternary alpha-substituted amino acids using memory of chirality has been developed. This strategy employs dynamic axial chirality of tertiary aromatic amides to memorize the initial chirality of an alpha-amino acid during the enolization step. Starting from L valine, an oxazolidin-5-one containing a tertiary aromatic amide was synthesized in one step and then alkylated with various electrophiles with good yield and enantioselectivity (up to 96%). Quaternary products can be obtained enantiomerically pure by recrystallization. One-step deprotection affords enantioenriched (S)-alpha-methyl valine (ee = 94%) or enantiopure (S)-alpha isopropyl aspartic acid (ee >99%) in only three steps starting from L-valine. PMID- 18410102 TI - A QM/MM approach to interpreting 67Zn solid-state NMR data in zinc proteins. AB - We present here a (67)Zn solid-state NMR investigation of Zn(2+) substituted rubredoxin. The sample has been prepared as both a dry powder and a frozen solution to determine the effects of static disorder on the NMR line shape. Low temperature experiments have been performed at multiple fields to determine the relative contributions to the NMR line shape from the electric field gradient and the anisotropic shielding tensors. Finally we present the theoretical interpretation of the experimental results utilizing a combined quantum mechanical molecular mechanics (QM/MM) approach. Theory predicts a sizable contribution from anisotropic shielding as compared with previously examined model systems. This is in good agreement with the experimental data. PMID- 18410103 TI - Sifuvirtide screens rigid membrane surfaces. establishment of a correlation between efficacy and membrane domain selectivity among HIV fusion inhibitor peptides. AB - Sifuvirtide, a 36 amino acid negatively charged peptide, is a novel and promising HIV fusion inhibitor, presently in clinical trials. Because of the aromatic amino acid residues of the peptide, its behavior in aqueous solution and the interaction with lipid-membrane model systems (large unilammelar vesicles) were studied by using mainly fluorescence spectroscopy techniques (both steady-state and time-resolved). No significant aggregation of the peptide was observed with aqueous solution. Various biological and nonbiological lipid-membrane compositions were analyzed, and atomic force microscopy was used to visualize phase separation in several of those mixtures. Results showed no significant interaction of the peptide, neither with zwitterionic fluid lipid membranes (liquid-disordered phase), nor with cholesterol-rich membranes (liquid-ordered phase). However, significant partitioning was observed with the positively charged lipid models (K(p) = (2.2 +/- 0.3) x 10(3)), serving as a positive control. Fluorescence quenching using Forster resonance acrylamide and lipophilic probes was carried out to study the location of the peptide in the membrane models. In the gel-phase DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) membrane model, an adsorption of the peptide at the surface of these membranes was observed and confirmed by using Forster resonance energy-transfer experiments. These results indicate a targeting of the peptide to gel-phase domains relatively to liquid-disordered or liquid-ordered phase domains. This larger affinity and selectivity toward the more rigid areas of the membranes, where most of the receptors are found, or to viral membrane, may help explain the improved clinical efficiency of sifuvirtide, by providing a local increased concentration of the peptide at the fusion site. PMID- 18410104 TI - Genetic engineering combined with deep UV resonance Raman spectroscopy for structural characterization of amyloid-like fibrils. AB - Elucidating the structure of the cross-beta core in large amyloid fibrils is a challenging problem in modern structural biology. For the first time, a set of de novo polypeptides was genetically engineered to form amyloid-like fibrils with similar morphology and yet different strand length. Differential ultraviolet Raman spectroscopy allowed for separation of the spectroscopic signatures of the highly ordered beta-sheet strands and turns of the fibril core. The relationship between Raman frequencies and Ramachandran dihedral angles of the polypeptide backbone indicates the nature of the beta-sheet and turn structural elements. PMID- 18410105 TI - The role of surface basal planes of layered mixed metal oxides in selective transformation of lower alkanes: propane ammoxidation over surface ab planes of Mo-V-Te-Nb-O M1 phase. AB - The surface ab planes of the M1 phase exposed selectively after atomic layer deposition (ALD) of alumina followed by crushing showed significantly improved selectivity to acrylonitrile during propane ammoxidation. The results demonstrated the importance of surface ab planes for the activity and selectivity of the M1 phase in propane ammoxidation and general utility of surface modification by ALD in studies of catalytic behavior of surface planes in layered mixed metal oxides. PMID- 18410107 TI - Engineering stacks of aromatic rings by the interpenetration of self-assembled coordination cages. AB - One-step quantitative self-assembly of aromatic towers consisting of seven to nine discretely stacked aromatic rings is achieved from 25-27 multicomponents via the interpenetration of two identical coordination cages. PMID- 18410108 TI - Metal-carbon nanotube contacts: the link between Schottky barrier and chemical bonding. AB - The field effect transistor based on carbon nanotubes (CNT) is a very promising candidate for post-CMOS microelectronics. Transport in the CNT channel is dominated by the Schottky barriers existing at the metal source contacts. The nature of the metal and the geometry of the contact appear to influence strongly the electrical behavior, but the mechanism is still rather obscure. Extensive calculations based on density functional theory performed for both end and side contacts and for two metals of very different nature, namely, Al and Pd, allow us to identify a clear connection between the character of the chemical bonding and the height of the Schottky barrier (SBH). Our results emphasize that a low SBH for hole conduction in a CNT implies that the pi-electron system of the latter is almost exclusively involved in the chemical bonding with the metal atoms at the interface and that the bonding is not too strong so that both orbital hybridization and topology are preserved. This is the case for Pd in both end and side configurations and to a large extent for Al but in the side geometry only. On the other hand, the coupling of the metal states with the sigma-like system or, in other words, the perturbation of the conjugation of the pi-system via sp3 C-hybridization is the mechanism that enhances the SBH. This is especially evident in the end contact with Al. By showing how the chemistry at the interfaces determines the SBH, our findings open the possibility of better controlling and designing "good contacts". PMID- 18410109 TI - Janus micelles induced by olefin metathesis. AB - A facile one-step procedure for hydrophobic modification and simultaneous TEM contrast enhancement via a regioselective olefin metathesis reaction using Grubbs' catalyst is presented. Polyether diblock copolymers were investigated, and both the chain ends of the hydrophilic and the hydrophobic block were hydrophobically modified. Modification of the hydrophilic block results in nonsymmetric supramolecular structures (Janus micelles) which self-assemble into larger hierarchically organized super-micelles. PMID- 18410111 TI - Considerations regarding the use of hyperspectral imaging data in classifications of food products, exemplified by analysis of maize kernels. AB - Development of robust analytical procedures is critical when using hyperspectral imaging technology in food technology and agriculture. This study used near isogenic inbred corn lines to address two basic questions: (1) To what extent is classification accuracy increased by grinding maize kernels? (2) Can the classification accuracy of two near-isogenic inbred lines be increased by using a spectral filter to classify only certain hyperspectral profiles from each image cube? Whole kernels and ground kernels in two particle intervals, 0.250-0.354 mm (size 1) and 0.354-0.841 mm (size 2), were examined. Spectral profiles acquired from ground kernels had higher spectral repeatability than data collected from whole kernels. The classification error of discriminant functions from whole kernels was >3 times lower than that of size 1 ground particles. Applying a spectral filter to input data had negligible effect on classifications of hyperspectral profiles from whole kernels and size 2 ground particles, but for size 1 ground particles a considerable increase in accuracy was observed. Independent validation confirmed that distinction between wild type and mutant inbred maize lines could be conducted with >80% accuracy after the proposed spectral filter had been applied to hyperspectral profiles of size 1 ground particles. A combination of discriminant analysis and regression analysis could be used to accurately predict mixture ratios of the two inbred lines. The use of spectral filtering to increase the level of spectral repeatability and the use of hyperspectral imaging technology in large-scale commercial operations are discussed. PMID- 18410112 TI - Synthesis of conjugated linoleic acid by human-derived Bifidobacterium breve LMC 017: utilization as a functional starter culture for milk fermentation. AB - This study was performed to discover bifidobacteria isolated from human intestines that optimally convert linoleic acid (LA) to conjugated linoleic acid (CLA) and to optimize the culture conditions of milk fermentation. One hundred and fifty neonatal bifidobacteria were screened for CLA-producing ability, and Bifidobacterium breve LMC 017 was selected as it showed about 90% conversion of free LA in MRS broth. The selected strain showed resistance at 0.5% LA in microaerophillic conditions. When monolinolein (LA 90%) was used as a substrate for CLA production, the conversion rate was lower compared to free LA, but the growth rate was unaffected during the milk fermentation. There was no significant difference in CLA production between aerobic and anaerobic conditions, and little decline in CLA was shown after the maximal CLA level had been reached. CLA production increased by 80% with 24 h of incubation in milk containing additional skim milk (5%), where the proteins may have facilitated the production of CLA by enhancing the interaction of substrate with the bacteria. CLA production did not decline after 9 h of fermentation and an additional 12 weeks of storage with other commercial starters. This demonstrates the possibility of using this strain as a costarter in the production of CLA-enriched yogurt. PMID- 18410113 TI - Zinc distribution and speciation within rocket plants (Eruca vesicaria L. Cavalieri) grown on a polluted soil amended with compost as determined by XRF microtomography and micro-XANES. AB - Zinc distribution and speciation within different organs (root, petiole, and leaf) of the edible plant Eruca vesicaria L. Cavalieri were determined using synchrotron microbeam X-ray techniques (XRF microtomography and mu-XANES) for plants grown in polluted soil with or without compost amendment. Data on soil derived from different extraction procedures and using mu-XANES analyses on rhizospheric soil indicated that compost amendment did not significantly influence the Zn speciation and availability in soil. However, major differences were observed within the plants. Plants grown in the presence of compost were able to partly block zinc immediately outside the root endodermis in the form of zinc-phytate, while a smaller Zn fraction was allowed to xylem transport as zinc citrate. In the leaves, zinc was largely excluded from leaf cells, and about approximately 50% was in the form of phosphate precipitates, and the other 50% was complexed by cysteine and histidine residues. The reported data provide new information concerning the mechanisms of zinc tolerance in E. vesicaria L. Cavalieri, a very common edible plant in Mediterranean regions, and on the role of compost in influencing the molecular strategies involved in zinc uptake and detoxification. PMID- 18410114 TI - Shining light on the differences in molecular structural chemical makeup and the cause of distinct degradation behavior between malting- and feed-type barley using synchrotron FTIR microspectroscopy: a novel approach. AB - The objective of this study was to use advanced synchrotron-sourced FTIR microspectroscopy (SFTIRM) as a novel approach to identify the differences in protein and carbohydrate molecular structure (chemical makeup) between these two varieties of barley and illustrate the exact causes for their significantly different degradation kinetics. Items assessed included (1) molecular structural differences in protein amide I to amide II intensities and their ratio within cellular dimensions, (2) molecular structural differences in protein secondary structure profile and their ratios, and (3) molecular structural differences in carbohydrate component peak profile. Our hypothesis was that molecular structure (chemical makeup) affects barley quality, fermentation, and degradation behavior in both humans and animals. Using SFTIRM, the protein and carbohydrate molecular structural chemical makeup of barley was revealed and identified. The protein molecular structural chemical makeup differed significantly between the two varieties of barleys. No difference in carbohydrate molecular structural chemical makeup was detected. Harrington was lower than Valier in protein amide I, amide II, and protein amide I to amide II ratio, while Harrington was relatively higher in model-fitted protein alpha-helix and beta-sheet, but lower in the others (beta turn and random coil). These results indicated that it is the molecular structure of protein (chemical makeup) that may play a major role in the different degradation kinetics between the two varieties of barleys (not the molecular structure of carbohydrate). It is believed that use of the advanced synchrotron technology will make a significant step and an important contribution to research in examining the molecular structure (chemical makeup) of plant, feed, and seeds. PMID- 18410115 TI - Fourier transform infrared spectra of drying oils treated by irradiation. AB - Drying oils, such as linseed oil and tung oil, have the potential as coating materials to improve barrier properties of biobased packaging films. Oil drying is a chemical reaction in which polyunsaturated fatty acids undergo autoxidation. During drying, oils polymerize and form water-resistant films. However, drying rates tend to be too slow for practical applications. Metal driers are used in the paint industry to accelerate drying, but often driers are not safe for food contact. The objective of this work was to investigate the effect of ionizing radiation on the oxidation or drying rate of drying oils. The effect of irradiation dose on the drying rate of linseed and tung oils was monitored by FTIR spectroscopy. The peak at 3010 cm (-1) was found to be a useful index of oxidation rate. The decrease in peak intensity with time was fitted with exponential functions of the form Abs = Abs 0 exp (- t/ k), where Abs 0 is the initial absorbance and 1/ k is the rate constant for the oxidation process. Values for k were 9.91 ( R (2) = 0.98), 6.59 ( R (2) = 0.95)n and 6.44 ( R (2) = 0.97) for radiation levels of 0, 50, and 100 kGy, respectively. The k values suggested that the oxidation rate increased as the radiation dose increased from 0 to 50 kGy. A further increase to 100 kGy had only a limited effect. PMID- 18410116 TI - Preparative procedures markedly influence the appearance and structural integrity of protein storage vacuoles in soybean seeds. AB - In legumes, vacuoles serve as the final depository for storage proteins. The protein storage vacuoles (PSVs) of soybean contain electron-transparent globoid regions in which phytic acid ( myo-inositol-1,2,3,4,5,6-hexakisphosphate) is sequestered. This paper reports the effect of preparative procedures on the appearance and ultrastructural integrity of PSVs in soybeans. Electron microscopy examination of both developing and mature soybean seeds that were postfixed with osmium tetroxide revealed PSVs that had a homogeneous appearance with very few globoid crystals dispersed in them. Numerous electron-dense lipid bodies were readily seen in these cells. Omission of osmium tetroxide strikingly altered the appearance of PSVs and aided the visualization of the location of the globoids in the PSVs. In contrast to the osmicated tissue, lipid bodies appeared as electron transparent spheres. The choice of dehydration reagent or staining procedure had little influence on the appearance of the PSVs. The results of this study demonstrate the profound effect of osmium tetroxide on the appearance and structural integrity of PSVs in soybean. PMID- 18410117 TI - Apigenin inhibits platelet adhesion and thrombus formation and synergizes with aspirin in the suppression of the arachidonic acid pathway. AB - Previous studies using washed platelets demonstrated that certain flavonoids inhibit platelet function through several mechanisms including blockade of TxA(2) receptors (TPs). We aimed to analyze the binding capacity of flavonoids to TPs in platelet rich plasma (PRP), investigated their effect in flowing blood, and evaluated the ability of apigenin to improve the efficacy of aspirin in the inhibition of platelet aggregation. The binding of flavonoids to TPs in PRP was explored using binding assays and the TP antagonist [ (3)H]SQ29548. Effects of flavonoids on platelet adhesion were assessed using arterial subendothelium with annular plate perfusion chambers, and global evaluation of apigenin on high-shear dependent platelet function was determined by the PFA-100. To evaluate the ability of apigenin to potentiate the effect of aspirin, arachidonic acid-induced platelet aggregation was measured prior to and after consumption of subaggregatory doses of aspirin in the presence or absence of apigenin. Binding assays revealed that apigenin was an efficient competitor of [ (3)H]SQ29548 binding to PRP ( K i = 155.3 +/- 65.4 microM), and perfusion studies showed that apigenin, genistein, and catechin significantly diminished thrombus formation when compared to control (26.2 +/- 3.8, 33.1 +/- 5.2, and 26.2 +/- 5.2 vs 76.6 +/ 2.6%, respectively; p < 0.05). Apigenin, similarly to the TP antagonist SQ29548, significantly prolonged collagen epinephrine-induced PFA-100 closure time in comparison to the control and, when added to platelets that had been exposed in vivo to aspirin, potentiated its inhibitory effect on platelet aggregation. The inhibitory effect of some flavonoids in the presence of plasma, particularly apigenin, might in part rely on TxA(2) receptor antagonism. There is a clear increase in the ex vivo antiplatelet effect of aspirin in the presence of apigenin, which encourages the idea of the combined use of aspirin and certain flavonoids in patients in which aspirin fails to properly suppress the TxA(2) pathway. PMID- 18410118 TI - Alkenol-alkyne cross metathesis. AB - Allyl alcohols and their homologues were used in the enyne cross metathesis to prepare hydroxy-functionalized dienes. An isomerization was found to occur under prolonged heating, and a method for conversion to ( E)-diene product is also reported. PMID- 18410119 TI - Molecular fibers and wires in solid-state and solution self-assemblies of cyclodextrin [2]rotaxanes. AB - Cyclodextrin [2]rotaxanes have been prepared by coupling dimethylanilines with dicarboxylic acids using DMT-MM, in aqueous solutions of alpha-cyclodextrin, and the example illustrated shows unusual fluorescence emission and other spectroscopic behavior characteristic of the formation of molecular wires in solution, similar to the fibers observed in the solid state. PMID- 18410120 TI - Cu2+-induced intermolecular static excimer formation of pyrenealkylamine. AB - Synthesis of monopyrenylalkylamine derivative 1 and its fluorescence behavior for Cu2+ in H2O/CH3CN (1:1, v/v) were investigated. Upon Cu2+ binding, 1, bearing a sulfonamide group, exhibited a marked excimer emission at 455 nm along with a weak monomer emission at 375 nm. The excimer emission, driven by formation of an intermolecular pyrenyl static excimer upon Cu2+ binding to the sulfonamide group, is rationalized by experimental and theoretical DFT calculation results. PMID- 18410121 TI - Stereocontrolled assembly of the C3/C3' dideoxy core of lomaiviticin A/B and congeners. AB - The dideoxy core (23) of lomaiviticinone and congener 15 were derived starting from (-)-quinic acid in a stereocontrolled fashion. PMID- 18410122 TI - Testing the validity of the conventional resonance model for protonated carbonyl, imine and thiocarbonyl compounds. An Ab initio valence bond study. AB - The conventional resonance model describes protonated carbonyls, imines, and thiocarbonyls by a superposition of two structures, one pi polar-covalent and the other of carbenium type. The validity of this model is clearly supported by high level valence bond calculations, giving a 32% weight for the carbenium form in protonated carbonyl, 19% in protonated formamine and thioformaldehyde. The carbenium form is further stabilized by pi-donating substituents. Solvation effects do not fundamentally change the gas-phase picture. PMID- 18410123 TI - Semisynthesis of dimeric proteins by expressed protein ligation. AB - A one-pot synthesis of homodimeric proteins is described. The synthetic strategy is based on a double expressed protein ligation reaction between thioester peptides and a new bis-cysteinyl linker. The protocol was also applied to the synthesis of heterodimers. PMID- 18410124 TI - Trinorlupeol: a major nonsterol triterpenoid in Arabidopsis. AB - We report the structure determination of 20,29,30-trinorlup-18-en-3beta-ol (trinorlupeol) and establish this novel C 27 metabolite as a major nonsterol triterpenoid in Arabidopsis thaliana. Trinorlupeol was concentrated in cuticular waxes, notably in the plant stem, floral buds, and seedpods, but not in leaves. Based on expression data and functional characterization of A. thaliana oxidosqualene cyclases, we propose that LUP1 is the cyclase responsible for trinorlupeol biosynthesis. Also described are two oxidized trinorlupeols and additional biosynthetic insights. PMID- 18410125 TI - Proline-catalyzed direct inverse electron demand diels-alder reactions of ketones with 1,2,4,5-tetrazines. AB - An organocatalytic direct inverse electron demand Diels-Alder reaction of ketones with 1,2,4,5-tetrazines has been developed. The process is efficiently catalyzed by proline to give Diels-Alder adducts pyridazines in high yields. PMID- 18410126 TI - Structure of Ca2+-bound S100A4 and its interaction with peptides derived from nonmuscle myosin-IIA. AB - S100A4, also known as mts1, is a member of the S100 family of Ca2+-binding proteins that is directly involved in tumor invasion and metastasis via interactions with specific protein targets, including nonmuscle myosin-IIA (MIIA). Human S100A4 binds two Ca2+ ions with the typical EF-hand exhibiting an affinity that is nearly 1 order of magnitude tighter than that of the pseudo-EF hand. To examine how Ca2+ modifies the overall organization and structure of the protein, we determined the 1.7 A crystal structure of the human Ca2+-S100A4. Ca2+ binding induces a large reorientation of helix 3 in the typical EF-hand. This reorganization exposes a hydrophobic cleft that is comprised of residues from the hinge region,helix 3, and helix 4, which afford specific target recognition and binding. The Ca2+-dependent conformational change is required for S100A4 to bind peptide sequences derived from the C-terminal portion of the MIIA rod with submicromolar affinity. In addition, the level of binding of Ca2+ to both EF hands increases by 1 order of magnitude in the presence of MIIA. NMR spectroscopy studies demonstrate that following titration with a MIIA peptide, the largest chemical shift perturbations and exchange broadening effects occur for residues in the hydrophobic pocket of Ca2+-S100A4. Most of these residues are not exposed in apo-S100A4 and explain the Ca2+ dependence of formation of theS100A4-MIIA complex. These studies provide the foundation for understanding S100A4 target recognition and may support the development of reagents that interfere with S100A4 function. PMID- 18410127 TI - Human replication protein A melts a DNA triple helix structure in a potent and specific manner. AB - Alternate DNA structures other than double-stranded B-form DNA can potentially impede cellular processes such as transcription and replication. The DNA triplex helix and G4 tetraplex structures that form by Hoogsteen hydrogen bonding are two examples of alternate DNA structures that can be a source of genomic instability. In this study, we have examined the ability of human replication protein A (RPA), a single-stranded DNA binding protein that is implicated in all facets of DNA metabolism, to destabilize DNA triplexes and tetraplexes. Biochemical studies demonstrate that RPA efficiently melts an intermolecular DNA triple helix consisting of a pyrimidine motif third strand annealed to a 4 kb duplex DNA fragment at protein concentrations equimolar to the triplex substrate. Heterologous single-stranded DNA binding proteins ( Escherichia coli SSB, T4 gene 32) melt the triplex substrate very poorly or not at all, suggesting that the triplex destabilizing effect of RPA is specific. In contrast to the robust activity on DNA triplexes, RPA does not melt intermolecular G4 tetraplex structures. Cellular assays demonstrated increased triplex DNA content when RPA is transiently repressed, suggesting that RPA melting of triple helical structures is physiologically important. On the basis of our results, we suggest that the abundance of RPA known to exist in vivo is likely to be a strong deterrent to the stability of triplexes that can potentially form from human genomic DNA sequences. PMID- 18410128 TI - Molecular dynamics simulations of human LRH-1: the impact of ligand binding in a constitutively active nuclear receptor. AB - The liver receptor homologue 1 (LRH-1 (NR5A2)) belongs to the orphan nuclear receptor family, indicating that initially no ligand was known. Although recent studies have shown that ligand binding can be obtained, the biological relevance remains elusive. Here, we modify the observed X-ray ligand into a biologically more significant phospholipid (phosphatidylserine, PS) present in human, to study, by molecular dynamics (MD) simulations, the impact of the ligand on the receptor and the interaction with different cofactor peptides. Furthermore, we characterize the interactions between receptor and the cofactor peptides of DAX-1 (NR0B1), Prox1 and SHP LXXLL box 1 and 2 (NR0B2) in terms of specificity. Our MD simulation results show different interaction patterns for the SHP box2 compared to DAX-1, PROX1 and SHP box1. SHP box2 shows specific interactions at its more C terminal end while the other investigated peptides show specific interactions at several positions but particularly at the +2 site. The peptide +2 side chain interacts with a charged amino acid of the receptor, in hLRH-1 Asp372. Together with the charge clamp residues Arg361 and Glu534, Asp372 forms a triangle shaped charge clamp responsible for peptide orientation and increased affinity. The binding of the PS ligand causes no overall structural changes of the receptor but affects the interactions with cofactor peptides. The cofactor peptides from SHP decrease its interaction with the receptor upon ligand binding while DAX-1 and PROX1 are unchanged or increase. The diverse ligand binding response of the cofactor provides an opportunity for drug design with the possibility to create agonist ligands to modify cofactor interaction. PMID- 18410130 TI - Intramolecular interactions in the N-domain of cardiac troponin C are important determinants of calcium sensitivity of force development. AB - Myocardial contraction is initiated when Ca2+ binds to site II of cardiac troponin C. This 12-residue EF-hand loop (NH2-DEDGSGTVDFDE-COOH) contains six residues (bold) that coordinate Ca2+ binding and six residues that do not appear to influence Ca2+ binding directly. We have introduced six single-cysteine substitutions (italics) within site II of cTnC to investigate whether these residues are essential for Ca2+ binding affinity in isolation and Ca2+ sensitivity of force development in single muscle fibers. Ca2+ binding properties of mutant proteins were examined in solution and after substitution into rat skinned soleus fibers. Except for the serine mutation, cysteine substitution had no effect on Ca2+ binding on cTnC in solution. However, as part of the myofilament, the threonine mutation reduced Ca2+ sensitivity while the phenylalanine mutation increased Ca2+ sensitivity. Analysis of the available crystal and NMR structures reveals specific structural mechanisms for these effects. PMID- 18410129 TI - The binding and release of oxygen and hydrogen peroxide are directed by a hydrophobic tunnel in cholesterol oxidase. AB - The usage by enzymes of specific binding pathways for gaseous substrates or products is debated. The crystal structure of the redox enzyme cholesterol oxidase, determined at sub-angstrom resolution, revealed a hydrophobic tunnel that may serve as a binding pathway for oxygen and hydrogen peroxide. This tunnel is formed by a cascade of conformational rearrangements and connects the active site with the exterior surface of the protein. To elucidate the relationship between this tunnel and gas binding and release, three mutant enzymes were constructed to block the tunnel or its putative gate. Mutation of the proposed gating residue Asn485 to Asp or tunnel residue Phe359 or Gly347 to Trp or Asn reduces the catalytic efficiency of oxidation. The K mO 2 increases from 300 +/- 35 microM for the wild-type enzyme to 617 +/- 15 microM for the F359W mutant. The k cat for the F359W mutant-catalyzed reaction decreases 13-fold relative to that of the wild-type-catalyzed reaction. The N485D and G347N mutants could not be saturated with oxygen. Transfer of hydride from the sterol to the flavin prosthetic group is no longer rate-limiting for these tunnel mutants. The steady state kinetics of both wild-type and tunnel mutant enzymes are consistent with formation of a ternary complex of steroid and oxygen during catalysis. Furthermore, kinetic cooperativity with respect to molecular oxygen is observed with the tunnel mutants, but not with the wild-type enzyme. A rate-limiting conformational change for binding and release of oxygen and hydrogen peroxide, respectively, is consistent with the cooperative kinetics. In the atomic resolution structure of F359W, the indole ring of the tryptophan completely fills the tunnel and is observed in only a single conformation. The size of the indole is proposed to limit conformational rearrangement of residue 359 that leads to tunnel opening in the wild-type enzyme. Overall, these results substantiate the functional importance of the tunnel for substrate binding and product release. PMID- 18410131 TI - High-throughput single copy DNA amplification and cell analysis in engineered nanoliter droplets. AB - A high-throughput single copy genetic amplification (SCGA) process is developed that utilizes a microfabricated droplet generator (microDG) to rapidly encapsulate individual DNA molecules or cells together with primer functionalized microbeads in uniform PCR mix droplets. The nanoliter volume droplets uniquely enable quantitative high-yield amplification of DNA targets suitable for long range sequencing and genetic analysis. A hybrid glass-polydimethylsiloxane (PDMS) microdevice assembly is used to integrate a micropump into the microDG that provides uniform droplet size, controlled generation frequency, and effective bead incorporation. After bulk PCR amplification, the droplets are lysed and the beads are recovered and rapidly analyzed via flow cytometry. DNA targets ranging in size from 380 to 1139 bp at single molecule concentrations are quantitatively amplified using SCGA. Long-range sequencing results from beads each carrying approximately 100 amol of a 624 bp product demonstrate that these amplicons are competent for achieving attomole-scale Sanger sequencing from a single bead and for advancing pyrosequencing read-lengths. Successful single cell analysis of the glyceraldehyde 3 phosphate dehydrogenase (GAPDH) gene in human lymphocyte cells and of the gyr B gene in bacterial Escherichia coli K12 cells establishes that SCGA will also be valuable for performing high-throughput genetic analysis on single cells. PMID- 18410132 TI - Analysis of glycopeptides using lectin affinity chromatography with MALDI-TOF mass spectrometry. AB - Glycopeptides prepared from 1 nmol of a mixture of glycoproteins, transferrin, and ribonuclease B by lysylendopeptidase digestion were isolated by lectin and cellulose column chromatographies, and then they were analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and MALDI-quadrupole ion trap (QIT)-TOF mass spectrometry which enables the performance of MS ( n ) analysis. The lectin affinity preparation of glycopeptides with Sambucus nigra agglutinin and concanavalin A provides the glycan structure outlines for the sialyl linkage and the core structure of N glycans. Such structural estimation was confirmed by MALDI-TOF MS and MALDI-QIT TOF MS/MS. Amino acid sequences and location of glycosylation sites were determined by MALDI-QIT-TOF MS/MS/MS. Taken together, the combination of lectin column chromatography, MALDI-TOF MS, and MALDI-QIT-TOF MS ( n ) provides an easy way for the structural estimation of glycans and the rapid analysis of glycoproteomics. PMID- 18410133 TI - Single-walled carbon nanotube network ultramicroelectrodes. AB - Ultramicroelectrodes (UMEs) fabricated from networks of chemical vapor deposited single-walled carbon nanotubes (SWNTs) on insulating silicon oxide surfaces are shown to offer superior qualities over solid UMEs of the same size and dimensions. Disk shaped UMEs, comprising two-dimensional "metallic" networks of SWNTs, have been fabricated lithographically, with a surface coverage of <1% of the underlying insulating surface. The electrodes are long lasting and give highly reproducible responses (either for repeat runs with the same electrode or when comparing several electrodes with the same size). For redox concentrations 14 A) showed residual fluorescence emission, while fluorescence quenching was observed for the shortest compounds. PMID- 18410136 TI - Relation of time of introduction of cow milk protein to an infant and risk of type-1 diabetes mellitus. AB - Several studies of infant feeding show a causal relationship between time of introduction of formula containing cow protein and risk of onset of type-1 diabetes mellitus. This paper cites the literature pro and con and discusses lipocalins which might play a role in the pathogensis. beta Lactoglobulin, a major lipocalin protein in bovine milk, is homologous to the human protein glycodelin (PP14), a T cell modulator. Anti-beta lactoglobulin cross-reacts with glycodelin. The newborn intestine does not have complete "closure" and can pass food antigens. Beta lactoglobulin could generate antibody to glycodelin undermining T cell regulation of beta cells. PMID- 18410137 TI - Influence of the protein staining in the fast ultrasonic sample treatment for protein identification through peptide mass fingerprint and matrix-assisted laser desorption ionization time of flight mass spectrometry. AB - The influence of the protein staining used to visualize protein bands, after in gel protein separation, for the correct identification of proteins by peptide mass fingerprint (PMF) after application of the ultrasonic in-gel protein protocol was studied. Coomassie brilliant blue and silver nitrate, both visible stains, and the fluorescent dyes Sypro Red and Sypro Orange were evaluated. Results obtained after comparison with the overnight in-gel protocol showed that good results, in terms of protein sequence coverage and number of peptides matched, can be obtained with anyone of the four stains studied. Two minutes of enzymatic digestion time was enough for proteins stained with coomassie blue, while 4 min was necessary when silver or Sypro stainings were employed in order to reach equivalent results to those obtained for the overnigh in-gel protein protocol. For the silver nitrate stain, the concentration of silver present in the staining solution must be 0.09% (w/v) to minimize background in the MALDI mass spectra. PMID- 18410139 TI - Physical properties of sequential interpenetrating polymer networks produced from canola oil-based polyurethane and poly(methyl methacrylate). AB - Sequential interpenetrating polymer networks (IPNs) were prepared using polyurethane (PUR) synthesized from canola oil-based polyol with terminal primary functional groups and poly(methyl methacrylate) (PMMA). The properties of the material were evaluated by dynamic mechanical analysis (DMA), differential scanning calorimetry (DSC), and modulated differential scanning calorimetry (MDSC), as well as tensile properties measurements. The morphology of the IPNs was investigated using scanning electron microscopy (SEM) and MDSC. A five-phase morphology, that is, sol phase, PUR-rich phase, PUR-rich interphase, PMMA-rich interphase, and PMMA-rich phase, was observed for all the IPNs by applying a new quantitative method based on the measurement of the differential of reversing heat capacity versus temperature from MDSC, although not confirmed by SEM, most likely due to resolution restrictions. NCO/OH molar ratios (cross-linking density) and compositional variations of PUR/PMMA both affected the thermal properties and phase behaviors of the IPNs. Higher degrees of mixing occurred for the IPN with higher NCO/OH molar ratio (2.0/1.0) at PUR concentration of 25 wt %, whereas for the IPN with lower NCO/OH molar ratio (1.6/1.0), higher degrees of mixing occurred at PUR concentration of 35 wt %. The mechanical properties of the IPNs were superior to those of the constituent polymers due to the finely divided rubber and plastic combination structures in these IPNs. PMID- 18410138 TI - Quantitative analysis of surface plasma membrane proteins of primary and metastatic melanoma cells. AB - Plasma membrane proteins play critical roles in cell-to-cell recognition, signal transduction and material transport. Because of their accessibility, membrane proteins constitute the major targets for protein-based drugs. Here, we described an approach, which included stable isotope labeling by amino acids in cell culture (SILAC), cell surface biotinylation, affinity peptide purification and LC MS/MS for the identification and quantification of cell surface membrane proteins. We applied the strategy for the quantitative analysis of membrane proteins expressed by a pair of human melanoma cell lines, WM-115 and WM-266-4, which were derived initially from the primary and metastatic tumor sites of the same individual. We were able to identify more than 100 membrane and membrane associated proteins from these two cell lines, including cell surface histones. We further confirmed the surface localization of histone H2B and three other proteins by immunocytochemical analysis with confocal microscopy. The contamination from cytoplasmic and other nonmembrane-related sources is greatly reduced by using cell surface biotinylation and affinity purification of biotinylated peptides. We also quantified the relative expression of 62 identified proteins in the two types of melanoma cells. The application to quantitative analysis of membrane proteins of primary and metastatic melanoma cells revealed great potential of the method in the comprehensive identification of tumor progression markers as well as in the discovery of new protein-based therapeutic targets. PMID- 18410140 TI - 2-chloro-(4R,5R)-bis[(1R,2S,5R)-menth-1-yloxycarbonyl)]-1,3,2-dioxaphospholane: a practical chiral pool-derived reagent for determining enantiomeric purity of alcohols. AB - 2-Chloro-(4 R,5 R)-bis[(1 R,2 S,5 R)-menth-1-yloxycarbonyl)]-1,3,2 dioxaphospholane is a practical reagent for reliably determining enantiomeric purity of chiral alcohols via (31)P NMR spectroscopy. The compound is available as a crystalline solid on a 20 g scale from PCl 3 and bis[(1 R,2 S,5 R)-menth-1 yl] tartrate. It is comparatively inert toward spontaneous hydrolysis under conventional laboratory conditions but undergoes quantitative substitution of alkoxide for chloride if treated with a chiral alcohol. Nonequivalent (31)P NMR signals of diastereomeric 2-alkoxy-1,3,2-dioxophospholanes were dispersed by approximately 1.4-0.1 ppm. The associated integral ratios reflected enantiomeric purities of preweighted samples of ( R)- and ( S)-1-phenylethanol, (+)- and (-) menthol, and a set of primary, secondary, and tertiary alcohols with a precision of +/-0.4-1.0%. PMID- 18410141 TI - Kinetics and mechanism of large rate enhancement in the alkaline hydrolysis of N' morpholino-N-(2'-methoxyphenyl)phthalamide. AB - The apparent second-order rate constant (k OH) for hydroxide-ion-catalyzed conversion of 1 to N-(2'-methoxyphenyl)phthalamate (4) is approximately 10(3) fold larger than k OH for alkaline hydrolysis of N-morpholinobenzamide (2). These results are explained in terms of the reaction scheme 1 --> k(1obs) 3 --> k(2obs) 4 where 3 represents N-(2'-methoxyphenyl)phthalimide and the values of k(2obs)/k(1obs) vary from 6.0 x 10(2) to 17 x 10(2) within [NaOH] range of 5.0 x 10(-3) to 2.0 M. Pseudo-first-order rate constants (k(obs)) for alkaline hydrolysis of 1 decrease from 21.7 x 10(-3) to 15.6 x 10(-3) s(-1) with an increase in ionic strength (by NaCl) from 0.5 to 2.5 M at 0.5 M NaOH and 35 degrees C. The values of k obs, obtained for alkaline hydrolysis of 2 within [NaOH] range 1.0 x 10(-2) to 2.0 M at 35 degrees C, follow the relationship k(obs) = kOH[HO(-)] + kOH'[HO (-)] (2) with least-squares calculated values of kOH and kOH' as (6.38 +/- 0.15) x 10(-5) and (4.59 +/- 0.09) x 10(-5) M (-2) s( 1), respectively. A few kinetic runs for aqueous cleavage of 1, N'-morpholino-N (2'-methoxyphenyl)-5-nitrophthalamide (5) and N'-morpholino-N-(2'-methoxyphenyl) 4-nitrophthalamide (6) at 35 degrees C and 0.05 M NaOH as well as 0.05 M NaOD reveal the solvent deuterium kinetic isotope effect (= k(obs) (H 2) (O)/ k(obs) (D 2 ) (O)) as 1.6 for 1, 1.9 for 5, and 1.8 for 6. Product characterization study on the cleavage of 5, 6, and N-(2'-methoxyphenyl)-4-nitrophthalimide (7) at 0.5 M NaOD in D2O solvent shows the imide-intermediate mechanism as the exclusive mechanism. PMID- 18410143 TI - Unsymmetric pyrrole, thiophene, and furan-conjugated comonomers prepared using azomethine connections: potential new monomers for alternating homocoupled products. AB - Unsymmetric comonomers consisting of thiophene, pyrrole, and furan heterocycles were prepared using azomethine bonds. Photophysical investigation of the novel pi donor-donor-donor segmented compounds revealed that their singlet excited state is only partially deactivated by internal conversion unlike their all-thiophene azomethine analogues. Temperature-dependent steady-state and time-resolved emission studies demonstrated that the unsymmetric compounds deactivated efficiently their singlet excited state by intersystem crossing to populate the triplet manifold. This lower energy state is rapidly deactivated by nonradiative self-quenching. The comonomers and their anodically prepared conjugated homocoupled products are both electrochemically active, resulting in new compounds that can be mutually oxidized and reduced. Meanwhile, the oxidation potentials of the coupled products are shifted by up to 400 mV to more cathodic potentials relative to their corresponding comonomers, confirming their increased degree of conjugation. PMID- 18410142 TI - New insight into Ni(II)-catalyzed cyclization reactions of propargylic compounds with soft nucleophiles: novel indenes formation. AB - We have disclosed a very nice advance of nickel(II)-catalyzed carboannulation reactions. Highly substituted indene derivatives are readily prepared in moderate to excellent yields under very mild reaction conditions in air via a nickel(II) catalyzed cyclization of propargylic compounds with soft nucleophiles. PMID- 18410144 TI - Density functional theory studies of negishi alkyl-alkyl cross-coupling reactions catalyzed by a methylterpyridyl-Ni(I) complex. AB - Density functional theory calculations were done to examine the potential energy surfaces of Ni(I)-catalyzed Negishi alkyl-alkyl cross-coupling reactions by using propyl iodide and isopropyl iodide as model alkyl electrophiles and CH 3ZnI as a model alkyl nucleophile. A four-step catalytic cycle involving iodine transfer, radical addition, reductive elimination, and transmetalation steps were characterized structurally and energetically. The reaction mechanism for this catalytic cycle appears feasible based on the calculated free energy profiles for the reactions. The iodine transfer step is the rate-determining step for the Ni(tpy)-CH 3 (tpy = 2,2'6',2''-terpyridine) reactions with alkyl iodides. For secondary alkyl electrophiles, the oxidative addition intermediate, Ni(III), prefers to undergo decomposition over reductive elimination, whereas for the primary alkyl electrophiles, Ni(III) prefers to undergo reductive elimination over decomposition based on comparison of the relative reaction rates for these two types of steps. In addition, thermodynamic data were employed to help explain why the yield of the coupled product is very low from the Ni(II)-alkyl halide reactions with organozinc reagents. PMID- 18410145 TI - New telluride-mediated elimination for novel synthesis of 2',3'-didehydro-2',3' dideoxynucleosides. AB - Several 2',3'-dideoxynucleosides (ddNs) and 2',3'-didehydro-2',3' dideoxynucleosides (d4Ns) are FDA-approved anti-HIV drugs. Via conveniently synthesized 2,2'-anhydronucleosides, we have developed a novel synthesis of d4Ns by discovering and applying a new telluride-mediated elimination reaction. Our experiment results show that after substitution of 2,2'-anhydronucleosides with a telluride monoanion, a telluride intermediate is formed, and its elimination leads to formation of the olefin products (d4Ns). Our mechanistic study indicates that this telluride-assisted reaction consists of two steps: substitution (or addition) and elimination. By using dimethyl ditelluride (0.1 equiv) as the reagent, d4Ns can be synthesized with yields up to 90% via this telluride mediated elimination. Our novel strategy has great potential to simplify synthesis of these drugs and to further reduce cost of AIDS treatment and will also facilitate development of novel d4N and ddN analogues. PMID- 18410146 TI - Total synthesis and determination of relative and absolute configuration of multiplolide A. AB - A flexible approach for total syntheses of possible multiplolide A diastereomers establishing the relative and absolute configuration is documented. The adopted strategy features ring-closing metathesis (RCM) as the key reaction and screening of a set of substrates for the feasibility of RCM in general and for the requisite E-configuration of ring olefin in particular. Selective protecting groups manipulation prior to the assembly of the central macrocyclic core was instrumental in installing the epoxide functionality on a fully deprotected nonenolide at the end of the synthesis. PMID- 18410147 TI - One-step synthesis of low molecular weight poly(p-phenyleneethynylenevinylene)s via polyaddition of aromatic diynes by catalysis of the [Ru(p-cymene)Cl2]2/AcOH system. AB - pi-Conjugated low molecular weight polymers characterized by regio- and stereoregular alternation of phenylene and ( E)-1-en-3-yne moieties have been synthesized by polyaddition of 1,4-diethynylbenzene or of 2,5-diethynyl-1,4 alkoxybenzene monomers, employing the commercially available di-micro-chlorobis[( p-cymene)chlororuthenium(II)] complex as the metal catalyst source, under homogeneous, atom-economical, amine- and phosphine-free conditions. Bulk materials of poly( p-phenyleneethynylenevinylene) derivatives are obtained with yields larger than 80%, from which polymers readily soluble in chlorinated solvents and in tetrahydrofuran are extracted in 60-75% yields. The polymers with average degrees of polymerization in the range n AV = 4-8 display optical properties in solution similar to those of the higher molecular weights analogues. PMID- 18410148 TI - A convenient method for the assignment of relative configuration of acyclic alpha alkyl-beta-hydroxy carbonyl compounds by (1)H NMR. AB - The relative configuration of acyclic alpha-alkyl-beta-hydroxy carbonyl compounds can be determined by using (1)H NMR spectroscopy. The assignment can be achieved by recording the (1)H NMR spectrum of the syn-anti mixture. The upfield carbinol hydrogen signal belongs to the anti whereas the downfield to the syn stereoisomer. PMID- 18410149 TI - Preparation of nonsymmetrically substituted stilbenes in a one-pot two-step heck strategy using ethene as a reagent. AB - We present here a strategy for the preparation of nonsymmetrically substituted stilbenes using a one-pot two-step double Heck strategy. First a protocol is developed for the selective preparation of a range of styrenes using ethene as the alkene coupling partner. Then conditions are found for the effective coupling of the styrenes with aryl halides using a 1:1 stoichiometric ratio of the two components. The use of the microwave apparatus to perform the reactions offers a convenient method for synthesis as well as for safely, easily, and accurately loading vessels with gaseous reagents. PMID- 18410150 TI - "Diversity oriented synthesis" of functionalized chiral tetrahydropyridines: potential GABA receptor agonists and azasugars from natural amino acids via a sequential Baylis-Hillman reaction and RCM protocol. AB - The preparation of chiral tetrahydropyridine-4-carboxylates as isoguvacine analogues and azasugars with a tertiary stereocenter from L-amino acids via diastereoselective a Baylis-Hillman reaction of N-allyl-Boc alpha-aminal, followed by ring-closing metathesis and dihydroxylation sequences, is reported. PMID- 18410152 TI - Antibacterial activity of fullerene water suspensions (nC60) is not due to ROS mediated damage. AB - The cytotoxic and antibacterial properties of nC 60, a buckminsterfullerene water suspension, have been attributed to photocatalytically generated reactive oxygen species (ROS). However, in this work, neither ROS production nor ROS-mediated damage is found in nC 60-exposed bacteria. Furthermore, the colorimetric methods used to evaluate ROS production and damage are confounded by interactions between nC 60 and the reagents, yielding false positives. Instead, we propose that nC 60 exerts ROS-independent oxidative stress, thus reconciling conflicting results in the literature. PMID- 18410151 TI - Selective human serum albumin sensor from the screening of a fluorescent rosamine library. AB - A fluorescent dye library approach for the development of a bioanalyte sensor was sought. The screening of a rosamine dye library against diverse macromolecules led to the discovery of a highly sensitive human serum albumin binder, G13, with approximately 36-fold fluorescence intensity change. G13 showed a highly selective response to HSA over other macromolecules including albumins from other species. The potential use of G13 for the detection of HSA in biofluids is described. PMID- 18410153 TI - Charge separation in type II tunneling structures of close-packed CdTe and CdSe nanocrystals. AB - We report on charge separation between type II aligned CdTe and CdSe nanocrystals. Two types of electrostatically bound nanocrystal structures have been studied: first, clusters of nanocrystals hold together by Ca(II) ions in aqueous solution and, second, thin film structures of nanocrystals created with layer-by-layer deposition in combination with polyelectrolytes. In both types of structures, short interparticle distances of less than 1 nm have been achieved, whereby the isolating organic ligands on the nanocrystal surfaces and/or the polymer monolayers act as tunneling barriers between nanocrystals. We have observed an efficient quenching of photoluminescence and a reduced emission lifetime for CdTe nanocrystals in both types of type II heterostructures. This result is explained by a spatial charge separation of the photoexcited electron hole pairs due to tunneling of charge carriers through the thin organic layer between CdTe and CdSe nanocrystals. Type II heterostructures demonstrated here may find future applications in photovoltaics. PMID- 18410154 TI - Emergence of ideal membrane cascades for downstream processing. AB - An algorithm is developed for describing ideal membrane cascades for fractionation of binary and pseudo-binary mixtures. It is shown that solvent management plays a key role in determining both purification and yield. Development of efficient diafilters is needed if membrane cascades are to achieve their full potential in competing with both chromatography and simulated moving bed operations in downstream processing of proteins. Such a replacement will also be important for fractionation of higher titers and larger substrates, such as plasmids, viruses, and even whole cells. PMID- 18410155 TI - A microscale yeast cell disruption technique for integrated process development strategies. AB - Miniaturizing protein purification processes at the microliter scale (microscale) holds the promise of accelerating process development by enabling multi-parallel experimentation and automation. For intracellular proteins expressed in yeast, small-scale cell breakage methods capable of disrupting the rigid cell wall are needed that can match the protein release and contaminant profile of full-scale methods like homogenization, thereby enabling representative studies of subsequent downstream operations to be performed. In this study, a noncontact method known as adaptive focused acoustics (AFA) was optimized for the disruption of milligram quantities of yeast cells for the subsequent purification of recombinant human papillomavirus (HPV) virus-like particles (VLPs). AFA operates by delivering highly focused, computer-controlled acoustic radiation at frequencies significantly higher than those used in conventional sonication. With this method, the total soluble protein release was equivalent to that of laboratory-scale homogenization, and cell disruption was evident by light microscopy. The recovery of VLPs through a microscale chromatographic purification following AFA treatment was within 10% of that obtained using homogenization, with equivalent product purity. The addition of a yeast lytic enzyme prior to cell disruption reduced processing time by nearly 3-fold and further improved the comparability of the lysate to that of the laboratory-scale homogenate. In addition, unlike conventional sonication methods, sample heating was minimized (< =8 degrees C increase), even using the maximum power settings required for yeast cell disruption. This disruption technique in combination with microscale chromatographic methods for protein purification enables a strategy for the rapid process development of intracellularly expressed proteins. PMID- 18410156 TI - Demonstration of robust host cell protein clearance in biopharmaceutical downstream processes. AB - Residual host cell protein impurities (HCPs) are a key component of biopharmaceutical process related impurities. These impurities need to be effectively cleared through chromatographic steps in the downstream purification process to produce safe and efficacious protein biopharmaceuticals. A variety of strategies to demonstrate robust host cell protein clearance using scale-down studies are highlighted and compared. A common strategy is the "spiking" approach, which is widely employed in clearance studies for well-defined impurities. For HCPs this approach involves spiking cell culture harvest, which is rich in host cell proteins, into the load material for all chromatographic steps to assess their clearance ability. However, for studying HCP clearance, this approach suffers from the significant disadvantage that the vast majority of host cell protein impurities in a cell culture harvest sample are not relevant for a chromatographic step that is downstream of the capture step in the process. Two alternative strategies are presented here to study HCP clearance such that relevance of those species for a given chromatographic step is taken into consideration. These include a "bypass" strategy, which assumes that some of the load material for a chromatographic step bypasses that step and makes it into the load for the subsequent step. The second is a "worst-case" strategy, which utilizes information obtained from process characterization studies. This involves operating steps at a combination of their operating parameters within operating ranges that yield the poorest clearance of HCPs over that step. The eluate from the worst case run is carried forward to the next chromatographic step to assess its ability to clear HCPs. Both the bypass and worst-case approaches offer significant advantages over the spiking approach with respect to process relevance of the HCP impurity species being studied. A combination of these small-scale validation approaches with large-scale HCP clearance data from clinical manufacturing and manufacturing consistency runs is used to demonstrate robust HCP clearance for the downstream purification process of an Fc fusion protein. The demonstration of robust HCP clearance through this comprehensive strategy can potentially be used to eliminate the need for routine analytical testing or for establishing acceptance criteria for these impurities as well as to demonstrate robust operation of the entire downstream purification process. PMID- 18410157 TI - Advances in primary recovery: centrifugation and membrane technology. AB - Significant and continual improvements in upstream processing for biologics have resulted in challenges for downstream processing, both primary recovery and purification. Given the high cell densities achievable in both microbial and mammalian cell culture processes, primary recovery can be a significant bottleneck in both clinical and commercial manufacturing. The combination of increased product titer and low viability leads to significant relative increases in the levels of process impurities such as lipids, intracellular proteins and nucleic acid versus the product. In addition, cell culture media components such as soy and yeast hydrolysates have been widely applied to achieve the cell culture densities needed for higher titers. Many of the process impurities can be negatively charged at harvest pH and can form colloids during the cell culture and harvest processes. The wide size distribution of these particles and the potential for additional particles to be generated by shear forces within a centrifuge may result in insufficient clarification to prevent fouling of subsequent filters. The other residual process impurities can lead to precipitation and increased turbidity during processing and even interference with the performance of the capturing chromatographic step. Primary recovery also poses significant challenges owing to the necessity to execute in an expedient manner to minimize both product degradation and bioburden concerns. Both microfiltration and centrifugation coupled with depth filtration have been employed successfully as primary recovery processing steps. Advances in the design and application of membrane technology for microfiltration and dead-end filtration have contributed to significant improvements in process performance and integration, in some cases allowing for a combination of multiple unit operations in a given step. Although these advances have increased productivity and reliability, the net result is that optimization of primary recovery processes has become substantially more complicated. Ironically, the application of classical chemical engineering approaches to overcome issues in primary recovery and purification (e.g., turbidity and trace impurity removal) are just recently gaining attention. Some of these techniques (e.g., membrane cascades, pretreatment, precipitation, and the use of affinity tags) are now seen almost as disruptive technologies. This paper will review the current and potential future state of research on primary recovery, including relevant papers presented at the 234th American Chemical Society (ACS) National Meeting in Boston. PMID- 18410158 TI - CLOPPA-IPPP analysis of cooperative effects in hydrogen-bonded molecular complexes. Application to intermolecular 2hJ(N,C) spin-spin coupling constants in linear (CNH)n complexes. AB - The cooperative effects on NMR indirect nuclear coupling constants are analyzed by means of the IPPP-CLOPPA approach (where CLOPPA is the Contributions from Localized Orbitals within the Polarization Propagator Approach and IPPP is the Inner Projections of the Polarization Propagator). The decomposition of the J coupling allows one to classify these effects as those due to changes in the geometric structure and those that directly involve the transmission mechanisms. This latter contribution admits a further classification, taking into account its electronic origin. As an example, the cooperative effects on intermolecular 2hJ(N,C) couplings of the linear complexes (CNH)n (n = 2, 3, 4) are discussed. PMID- 18410159 TI - Unfolded protein and peptide dynamics investigated with single-molecule FRET and correlation spectroscopy from picoseconds to seconds. AB - Single-molecule fluorescence spectroscopy and correlation methods are finding increasing applications in the investigation of biomolecular dynamics, especially together with Forster resonance energy transfer (FRET). Here, we use the combination of start-stop experiments and classical fluorescence correlation spectroscopy (FCS) to obtain complete intensity auto- and cross-correlation functions from picoseconds to seconds for investigating the dynamics of unfolded proteins and peptides. In combination with distance information from single molecule transfer efficiency histograms, we can analyze the data in terms of a diffusive process on a potential of mean force to obtain intramolecular diffusion coefficients. This allows us to extend our previous analysis of the time scales of chain dynamics into the low nanosecond range for peptides and into the microsecond range for a small cold shock protein (Csp). Dynamics in short unstructured peptides can be detected down to a time scale of about 10 ns, placing a lower limit on the time scales accessible with correlation methods and currently used dye pairs. We find no evidence for microsecond fluctuations in unfolded Csp, suggesting that its global chain dynamics occur predominantly in the tens of nanosecond range. We further investigate the position dependence of these dynamics by placing donor and acceptor dyes at different positions within the chain and find a decrease in the intramolecular diffusion coefficient by a factor of 3 upon moving one of the dyes toward the center of the polypeptide. Obtaining dynamic information on a wide range of time scales from single-molecule photon statistics will be of increasing importance for the study of unfolded proteins and for biomolecules in general. PMID- 18410160 TI - Effect of alpha-lactalbumin on aerosol-OT phase structures in oil/water mixtures. AB - The ability of water-soluble, globular proteins to tune surfactant/oil/water self assemblies has potential for the formation of biocompatible microemulsions and also plays a role in protein function at biological interfaces. In this work, we examined the effect of the protein alpha-lactalbumin on Aerosol-OT (AOT) phase structures in equivolume mixtures of oil and 0.1 M brine. In this pseudo-ternary system, surfactants are free to move to either oil or water phase to adopt phase structures close to the spontaneous curvature of the surfactants. Using small angle X-ray scattering, we observed that addition of this protein changed the spontaneous curvature of the surfactant monolayer substantially. In the absence of protein, AOT adopted a negative spontaneous curvature to form spherical w/o microemulsion droplets. When less than 1 wt % of alpha-lactalbumin was added into the system, the w/o droplets became nonspherical and larger in volume, corresponding to an increase in water uptake into the droplets. As the protein-to surfactant ratio increased, protein, surfactant, and oil increasingly partitioned toward the aqueous phase. There the protein triggered the formation of o/w microemulsions with a positive spontaneous curvature. These protein-containing structures exhibited significant interparticle attraction. We also compared the influence of two oil types, isooctane and cyclohexane, on the protein/surfactant interactions. We propose that the more negative natural curvature of the AOT/cyclohexane monolayer in the absence of protein prevented protein incorporation within organic phase structures and consequently pushed the system self-assembly toward aqueous aggregate formation. PMID- 18410161 TI - Magnetic and optical properties of Cu(II)-bis(oxamato) complexes: combined quantum chemical density functional theory and vibrational spectroscopy studies. AB - Vibrational spectroscopies are shown to be highly sensitive to the structural modifications of paramagnetic mono- and trinuclear Cu(II)-bis(oxamato) complexes. The vibrational bands are assigned using density functional theory (DFT) calculations. Moreover, Raman spectroscopy investigations for different temperatures of thin films show that the onset of superexchange interactions at low temperatures does not involve a modification of the structural parameters. The influence of packing effects, however, on the magnetic properties is significant, as demonstrated by means of DFT using the broken symmetry approach. PMID- 18410162 TI - Analysis and optimization of saturation transfer difference NMR experiments designed to map early self-association events in amyloidogenic peptides. AB - Saturation transfer difference (STD) methods recently have been proposed to be a promising tool for self-recognition mapping at residue and atomic resolution in amyloidogenic peptides. Despite the significant potential of the STD approach for systems undergoing oligomer/monomer (O/M) equilibria, a systematic analysis of the possible artifacts arising in this novel application of STD experiments is still lacking. Here, we have analyzed the STD method as applied to O/M peptides, and we have identified three major sources of possible biases: offset effects, intramonomer cross-relaxation, and partial spin-diffusion within the oligomers. For the purpose of quantitatively assessing these artifacts, we employed a comparative approach that relies on 1-D and 2-D STD data acquired at different saturation frequencies on samples with different peptide concentrations and filtration states. This artifact evaluation protocol was applied to the Abeta(12 28) model system, and all three types of artifacts appear to affect the measured STD spectra. In addition, we propose a method to minimize the biases introduced by these artifacts in the Halpha STD distributions used to obtain peptide self recognition maps at residue resolution. This method relies on the averaging of STD data sets acquired at different saturation frequencies and provides results comparable to those independently obtained through other NMR pulse sequences that probe oligomerization, such as nonselective off-resonance relaxation experiments. The artifact evaluation protocol and the multiple frequencies averaging strategy proposed here are of general utility for the growing family of amyloidogenic peptides, as they provide a reliable analysis of STD spectra in terms of polypeptide self-recognition epitopes. PMID- 18410163 TI - Facile fabrication of core-in-shell particles by the slow removal of the core and its use in the encapsulation of metal nanoparticles. AB - Core-in-shell particles with controllable core size have been fabricated from core-shell particles by means of the controlled core-dissolution method. These cores in inorganic shells were employed as scaffolds for the synthesis of metal nanoparticles. After dissolution of the cores, metal nanoparticles embedded in cores were encapsulated into the interior of shell, without any damage or change. This article describes a very simple method for deriving core-in-shell particles with controllable core size and encapsulation of nanoparticles into the interior of shell. PMID- 18410164 TI - Bilateral neurotoxic amygdala lesions in rhesus monkeys (Macaca mulatta): consistent pattern of behavior across different social contexts. AB - Although the amygdala has been repeatedly implicated in normal primate social behavior, great variability exists in the specific social and nonsocial behavioral changes observed in nonhuman primates with bilateral amygdala lesions. One plausible explanation pertains to differences in social context. This study measured the social behavior of amygdala-lesioned and unoperated rhesus monkeys (Macaca mulatta) in 2 contexts. Monkeys interacted in 4-member social groups over 32 test days. They were previously assessed in pairs (N. J. Emery et al., 2001) and were therefore familiar with each other at the beginning of this study. Across the 2 contexts, amygdala lesions produced a highly consistent pattern of social behavior. Operated monkeys engaged in more affiliative social interactions with control partners than did controls. In the course of their interactions, amygdala-lesioned monkeys also displayed an earlier decrease in nervous and fearful personality qualities than did controls. The increased exploration and sexual behavior recorded for amygdala-lesioned monkeys in pairs was not found in the 4-member groups. The authors concluded that the amygdala contributes to social inhibition and that this function transcends various social contexts. PMID- 18410166 TI - Sleep-dependent learning and practice-dependent deterioration in an orientation discrimination task. AB - Learning new information requires practice. The degree of learning can be influenced by the amount of practice and whether subjects sleep between sessions. Over-practice, however, can lead to performance deterioration. The interaction between practice-dependent deterioration and sleep-dependent learning needs more study. We examine whether the amount of practice before sleep alters learning, and whether prior sleep protects against deterioration. Two groups (N = 33) were tested three times across two days on an orientation discrimination task. The High practice group was tested twice before a night of sleep and once after, at 9 a.m., 7 p.m., and 9 a.m. The Low practice group was tested once before a night of sleep and twice after, at 7 p.m., 9 a.m., and 7 p.m. Overall, both groups showed (1) deterioration with repeated, within-day testing, (2) performance improvement only after a night of sleep, (3) similar amounts of sleep-dependent learning and practice-dependent deterioration. In summary, we found that sleep resets visual contrast thresholds to a lower baseline (i.e., produces learning), but does not prevent over-practice deterioration effects. Likewise, over-practice deterioration does not influence the magnitude of overnight learning on this task. PMID- 18410167 TI - Masculinized finger-length ratios of boys, but not girls, are associated with attention-deficit/hyperactivity disorder. AB - Gonadal hormones may exert permanent organizational effects on sexually dimorphic finger-length ratios and sexually dimorphic behavior expressed in childhood attention deficit-hyperactivity disorder (ADHD). This study extended recent work examining associations between finger-length ratios (specifically, 2D:4D) and ADHD in a well-characterized, clinically diagnosed, community-recruited sample of boys and girls. A multistage, diagnostic procedure was utilized to identify 113 children with ADHD and 137 non-ADHD comparison children. Right-hand digit ratios showed significant mean differences by gender, as well as associations with ADHD diagnosis. Boys with ADHD had more masculinized digit ratios than control-group boys. More masculine right 2D:4D and 3D:4D ratios were correlated with parent- and teacher-rated inattentive and hyperactive-impulsive symptoms in boys but not in girls. Masculinized finger-length ratios were associated with hyperactive impulsive and oppositional- defiant symptoms, but associations were largest with symptoms of inattention. It is concluded that prenatal, organizational effects of gonadal hormones may play a role in the development of ADHD and contribute to explaining sex differences in the prevalence rates of this childhood disorder. PMID- 18410168 TI - Chronic immobilization stress alters aspects of emotionality and associative learning in the rat. AB - Chronic stress significantly alters limbic neuroarchitecture and function, and potentiates emotionality in rats. Chronic restraint stress (CRS) increases aggression among familiar rats, potentiates anxiety, and enhances fear conditioning. Chronic immobilization stress (CIS) induces anxiety behavior and dendritic hypertrophy in the basolateral amygdala, which persist beyond a recovery period. However, little else is known about the emotional impact of CIS as a model of chronic stress or depression. Therefore, the authors present two experiments examining emotional and learned responses to CIS. In Experiment I, the authors examine individual differences in behaviors during and after CIS, specifically: struggling, aggression, learned helplessness, inhibitory avoidance, and escape behavior. In Experiment II, the authors confirm the effects of CIS on aggression and struggling during immobilization, and correlate individual responses with aspects of conditioned fear. Here the authors report significant effects of CIS on aggression, inhibitory avoidance, escape, as well as learned aspects of fear (i.e., fear conditioning) and inescapable stress (i.e., struggling and helplessness). These results emphasize the emotional and learned responses to CIS evident during and after the stress treatment, as well as the importance of individual differences. PMID- 18410169 TI - Evidence for social anxiety and impaired social cognition in a mouse model of fragile X syndrome. AB - This study assessed social behavior in a mouse model of Fragile X syndrome (FXS), the Fmr1 (tm1Cgr) or Fmr1 "knockout" (KO) mouse. Both the KO and wild-type (WT) mice preferred to be near a novel conspecific than to be alone. However, during the initial interaction with a novel conspecific, (1) a greater proportion of the KO mice exhibited high levels of grooming; and (2) the average duration of nose contact with the stimulus mouse was significantly shorter for the KO mice, both indicative of increased arousal and/or anxiety. Both groups exhibited a robust novelty preference when the novel animal was a "preferred" mouse. However, when the novel mouse was a "nonpreferred" animal, both groups showed a diminished novelty preference but this effect was more pronounced for the WT mice. This blunted negative reaction of the KO mice to a nonpreferred animal may indicate that they were less proficient than controls in distinguishing between positive and negative social interactions. These findings provide support for the use of this animal model to study the autistic features of FXS and autism spectrum disorders. PMID- 18410170 TI - 17beta-estradiol modifies stress-induced and age-related changes in hippocampal synaptic plasticity. AB - The female steroid hormone 17beta-estradiol enhances synaptic transmission and the magnitude of longterm potentiation (LTP) in adult rodent hippocampal slices. Long-term depression (LTD), another form of synaptic plasticity, occurs more prominently in hippocampal slices from aged rodents. A decrease in LTP has been recorded in hippocampal slices from adult rodents behaviorally stressed just before tissue preparation and electrophysiological recording. Here, the authors test the hypothesis that estrogen modifies synaptic plasticity in both adult and aged rodents, whether behaviorally stressed or not. Our results indicate that estrogen enhances LTP and attenuates LTD, thus producing a protective effect against both aging and stress. These results also provide new approaches that can be used to reverse age and stress-related learning and memory dysfunction. PMID- 18410171 TI - Maternal care, isolation-induced infant ultrasonic calling, and their relations to adult anxiety-related behavior in the rat. AB - In the rat, variations in maternal care affect the development of stable individual differences in anxiety-related behavior. Here, it was asked whether such experience-dependent differences can be detected already during early life. As a measure for anxiety in pups, isolation-induced ultrasonic vocalizations were used, and their dependency on different maternal behaviors, namely licking, retrieval behavior, and responsiveness to playback of pup calls, was tested. Consistent with reported differences of adult rats with high or low levels of maternal care experienced, the rarely licked offspring appeared to be more anxious, since they emitted more calls when separated from their mother and litter. Based on these findings, it was examined whether infant calling can be used as a predictor of adult anxiety-related behavior. Results show that infant call emission was negatively correlated with immobility and calling during fear conditioning. These relationships seem to be mediated at least partly by maternal care. In total, measuring ultrasonic vocalizations can provide information about an affective trait of infant and adult rats, which gives the opportunity to study the development of emotionality from early life onward. PMID- 18410172 TI - NF-kappaB regulates prenatal stress-induced cognitive impairment in offspring rats. AB - The study was designed to investigate whether nuclear factor of kappa B (NF kappaB) plays regulating role in prenatal stress-induced cognitive impairment and oxidative damage in offspring rats. The authors used a rat model to study plasma levels of corticosterone and oxidative DNA damage (8-OH-dG), protein expression of P65/p50 NF-kappaB, and cognitive function in female and male offspring rats in middle pregnant stage and later pregnant stage. Prenatal stress affected the capability of learning and memory in the offspring, especially in later stage stressed female offspring. The levels of corticosterone and 8-OH-dG were enhanced in response to stress. Both middle and later stage stresses induced a significant decrease in P65 expression and a significant increase in P50 expression in female offspring. In addition, later stage stress induced a significant decrease in P50 expression in male offspring. These results suggest that NF-kappaB complex may be acting in a positive regulatory fashion in prenatal stress-induced cognitive impairment and that oxidative DNA damage may exacerbate the activation of NF kappaB. PMID- 18410173 TI - Advancing the spontaneous hypertensive rat model of attention deficit/hyperactivity disorder. AB - To advance the spontaneous hypertensive rat (SHR) model of attention deficit/hyperactivity disorder (ADHD), experiments examined the SHR in tasks recognized to assess functioning of the prefrontal cortex or dorsal striatal. Tasks included odor-delayed win-shift (nonspatial working and reference memory), win-stay (habit learning), and attentional set-shifting (attention and behavioral flexibility). In Experiment 1, the SHR strain was compared with Wistar-Kyoto (WKY) and Wistar-Kyoto Hypertensive (WKHT) strains on the first 2 tasks. In Experiment 2, oral methylphenidate (1.5 mg/kg) and vehicle (water) were evaluated on all 3 tasks in SHR and WKY strains. Results demonstrated that the SHR made significantly more errors in the odor-delayed win-shift, win-stay, and attentional set-shifting tasks compared with the WKY. Similar performances in the WKY and WKHT indicated that deficits observed in the SHR were not related solely to hypertension. Treating the SHR with methylphenidate eliminated strain differences in all 3 tasks. These findings provide evidence that the SHR is a valid model for studying ADHD-associated neurocognitive deficits. Moreover, the current behavioral approach is appropriate to assess novel medications developed to target ADHD-associated neurocognitive deficits. PMID- 18410174 TI - Real-time dopamine efflux in the nucleus accumbens core during Pavlovian conditioning. AB - To assess the role of dopamine input to the nucleus accumbens core in anticipatory learning, fast-scan cyclic voltammetry was combined with appetitive Pavlovian conditioning. One group of rats (Paired) received 16 tone-food pairings for at least four daily sessions while the control group (Unpaired) received the same number of unpaired tone and food presentations. Both groups showed transient dopamine responses during food presentation throughout training, confirming dopamine involvement in reward processing. Only the Paired Group, however, showed consistently timed dopamine transients during the 10-s tone presentation. Transients first appeared near the end of the tone period as each animal acquired the tone-food association and then occurred progressively sooner on subsequent sessions. Later sessions also revealed a consistently timed dopamine response soon after food delivery in Paired animals. Collectively, these results implicate phasic dopamine release in the acquisition of Pavlovian learning and also suggest an early dopamine response to the unconditioned stimulus as training continues. PMID- 18410175 TI - The roles of accumbal dopamine D1 and D2 receptors in maternal memory in rats. AB - Female rats show enhanced maternal responsiveness toward their young if they have had maternal experiences before. This kind of maternal experience-based memory is critically dependent on the mesolimbic dopamine (DA) system, especially the nucleus accumbens (NA) shell. However, the relative contributions of the two main DA receptor systems (D1 and D2) within the shell have not been delineated. This study investigates the roles of dopamine D1 and D2 receptors in maternal memory by infusing a selective D1 antagonist, SCH-23390; a selective D2 antagonist, sulpiride; or a combination D-1/D-2 antagonist, cis-Z-flupenthixol, into the NA shell of postpartum female rats. Sulpiride-infused rats showed a significantly longer latency to exhibit full maternal behavior following a 10-day pup isolation period in comparison to the controls that received a vehicle. Cis-Z-flupenthixol disrupted maternal memory to a greater extent, as rats receiving this showed the longest latencies to express maternal behavior. SCH-23390 infusions had only marginal effects. These findings suggest that both the D1 and the D2 receptor subtypes play a role in the consolidation of maternal memory and they might do so by mediating the motivational salience of pup stimulation. PMID- 18410176 TI - Differential effects of dopamine and AMPA receptor antagonists on the expression of conditioned avoidance responding in rats. AB - AMPA receptor antagonists disrupt avoidance responding, but their day-to-day effect on this behavior has not been elucidated. This study compared the multisession effect of the AMPA/kainate receptor antagonist CNQX with that of the typical antipsychotic haloperidol on the expression of avoidance responding. Rats (N = 199) were trained to move to safety on presentation of a tone in one-way active conditioned avoidance and were tested across 5 sessions. Intracerebroventricular (icv) injection of CNQX (20-min injection-test interval) produced a dose-dependent, immediate block of avoidance responding, compared with the extinction-like decline of avoidance responding produced by haloperidol (intraperitoneal [ip], 60-min injection-test interval; icv, 60 but not 20-min injection-test interval). Previous exposure to CNQX significantly reduced its efficacy, illustrating that its effects may not be specific to the conditioned safety-related stimuli that control responding in conditioned avoidance, as proposed for antidopaminergic compounds. The new multisession profile of disrupted avoidance responding illustrated by CNQX suggests different roles for glutamatergic and dopaminergic neurotransmission in conditioned avoidance responding. Results are consistent with a role for AMPA receptors in maintaining the expression of learning. PMID- 18410177 TI - Neurochemical basis of conditioned partner preference in the female rat: I. Disruption by naloxone. AB - The effects of the opioid antagonist naloxone were examined on the development of conditioned partner preference induced by paced copulation in female rats. In Experiment 1, ovariectomized, hormone-primed rats were conditioned to associate scented and unscented male rats with paced and nonpaced copulation, respectively. Female rats in Experiment 2 associated albino or pigmented male rats with paced or nonpaced copulation. Naloxone or saline was administered before each conditioning trial. During a final drug-free preference test, female rats could choose to copulate with either a pacing related or unrelated male. Saline-trained female rats in the paired group copulated preferentially with the pacing-related male rat, whereas naloxone-trained female rats did not show a preference. The authors concluded that opioids mediated the conditioned partner preference induced by paced copulation. PMID- 18410178 TI - Neurochemical basis of conditioned partner preference in the female rat: II. Disruption by flupenthixol. AB - The effects of the dopamine receptor antagonist flupenthixol were examined on the development of conditioned partner preference induced by paced copulation in female rats. In Experiment 1, ovariectomized, hormone-primed rats were conditioned to associate scented and unscented males with paced and nonpaced copulation, respectively. Females in Experiment 2 associated albino or pigmented males with paced or nonpaced copulation. Flupenthixol or saline was administered before each conditioning trial. During a final drug-free preference test, females could choose to copulate with either a pacing-related or nonpacing-related male. Saline-trained females copulated preferentially with the pacing-related male, whereas flupenthixol disrupted odor but not strain conditioning. The role of dopamine in conditioned partner preference depends on the type of stimuli to be learned. PMID- 18410180 TI - Sex versus sweet: opposite effects of opioid drugs on the reward of sucrose and sexual pheromones. AB - Endogenous opioids mediate some reward processes involving both natural (food, sweet taste) and artificial (morphine, heroin) rewards. In contrast, sexual behavior (which is also reinforcing) is generally inhibited by opioids. To establish the role of endogenous opioids for a newly described natural reinforcer, namely male sexual pheromones for female mice, we checked the effects of systemic injections of the general opioid antagonist naloxone (1-10 mg/kg) and the agonist fentanyl (0.1- 0.5 mg/kg) in a number of behavioral tests. Naloxone affected neither the innate preference for male-soiled bedding (vs. female-soiled bedding) in 2-choice tests nor the induction of place conditioning using male pheromones as rewarding stimuli, although it effectively blocked the preference for consuming a sucrose solution. In contrast, fentanyl inhibited the preference for male chemosignals without altering sucrose preference. These results suggest that, in macrosmatic animals such as rodents, opioidergic inhibition of sexual behavior might be due, at least partially, to an impaired processing of pheromonal cues and that the hedonic value of sweet-tasting solutions and sexual pheromones are under different opioid modulation. PMID- 18410181 TI - Fish on Prozac: effect of serotonin reuptake inhibitors on cognition in goldfish. AB - The authors studied the effect of fluoxetine (a specific serotonin reuptake inhibitor--SSRI) on active avoidance learning in fish. In an active, two-way, shuttle-box avoidance task we compared escape and avoidance of shock among fish receiving chronic administration of fluoxetine (Prozac), others receiving a 5 HT(1A) receptor antagonist (WAY 100,635; Sigma, St. Louis, MO) and controls. The receptor-blocked group performed significantly more Avoids than the fluoxetine group, which was lowest in performance, or than controls, which were slightly higher, and not significantly different from, the fluoxetine group. This conforms to results reported in rats. Active avoidance learning may be stimulated by the 5 HT(1A) receptor antagonist. Fluoxetine-treated fish seemed less active in their home tanks than controls or blocker-treated fish. These results suggest that at the dosages used in this experiment, the 5-HT(1A) receptor antagonist WAY 100,635 may have positive effects on cognition in fishes and its action may not necessarily be restricted to blockage of 5 HT reuptake inhibition. It seems that serotonin mechanisms may be highly conserved in vertebrate evolution. PMID- 18410179 TI - Investigating the motivational mechanism of altered saline consumption following 5-HT(1A) manipulation. AB - The precise role played by serotonin (5-HT) in taste--an issue of great interest given the involvement of serotonin in human sensory and eating disorders--is a matter of considerable debate, perhaps because of the variety of methodologies that have been brought to bear by different researchers. Here, we use multiple methods to reveal the motivational mechanism whereby 5-HT(1A) receptor activation modulates drinking behavior. Subcutaneous injections of the selective 5-HT(1A) agonist 8-hydroxy-2-di-n-propylamino-tetralin (8-OH-DPAT), a drug that reduces 5 HT release by acting on presynaptic autoreceptors, dose-dependently increased consumption of 0.45 M NaCl in a one-bottle test. In a two-bottle test, however, 8 OH-DPAT-treated animals (30 microg/kg/ml) demonstrated decreased NaCl preference- although our detection of this effect was obscured by adaptation to the drug across days. Rats' performance in a brief access test confirmed that 8-OH-DPAT decreased preference for saline by both increasing water consumption and decreasing NaCl consumption. Finally, taste reactivity tests demonstrated that the latter result does not reflect decreased NaCl palatability. Overall, the results suggest that 8-OH-DPAT-induced 5-HT hypofunction increases thirst without substantially affecting the palatability of NaCl. PMID- 18410182 TI - Iron-induced oxidative stress modulates olfactory learning and memory in honeybees. AB - Reactive oxygen species (ROS)-mediated oxidative stress tends to increase with environmental stress, aging, and age-related diseases resulting in progressive neuronal dysfunction. The purpose of the present study was to examine whether or not oxidative stress can be induced into the antennal lobes of the honeybee brain by injecting ferrous ammonium citrate (FAC). Proboscis Extension Reflex conditioning procedure was used to assay subjects' responses to odorants for evaluating the effect of oxidative stress on the olfactory learning and memory. FAC-induced inhibitory effect on olfactory learning and memory was dose-and time dependent. Injections of reduced glutathione (GSH) into the antennal lobes before FAC treatment blocked oxidative stress-mediated inhibitory effect. Injections of VK-28 prior to FAC treatment overcame oxidative stress-mediated inhibitory response. However, injections of GSH into the antennal lobes prior to mianserin/dsRNA treatment did not reverse octopamine receptor disruption-mediated inhibitory response. These results indicate that normal cellular redox is crucial for olfactory processing, and chelation of iron prevents ROS-mediated oxidative stress. Furthermore, octopamine receptor disruption, and FAC-mediated oxidative stress confer two independent mechanisms that impair olfactory learning and memory in honeybees. PMID- 18410183 TI - Locomotor and exploratory behavior in the rat following bilateral vestibular deafferentation. AB - Despite many studies of the postural and ocular reflex deficits caused by chronic bilateral vestibular loss in rats and guinea pigs, there have been few systematic studies of the effects of vestibular loss on locomotor activity and exploratory behavior over a period of several months following the lesion. In this study, the authors quantified locomotor and exploratory behavior in an open field maze at 3 weeks, 3 months, and 5 months following bilateral vestibular loss in rats. As a result of bilateral surgical vestibular lesions, rats exhibited a persistent increase in locomotor velocity, duration, and distance traveled, with a marked tendency for increased inner field activity and reduced thigmotaxis. Rats without balance-sense were also found to spend less time exploring the environment, as indicated by a decreased frequency and duration of wall-supported rearings. These results suggest that sudden and complete loss of balance-sense has persistent and complex effects on the way that rats navigate through and explore the environment. PMID- 18410184 TI - Delayed extinction and stronger reinstatement of cocaine conditioned place preference in adolescent rats, compared to adults. AB - Adolescence is a transitional period during development that is associated with a greater likelihood of addiction to drugs than any other age. One possibility for this observation is that learned associations between the rewarding experience of drugs and drug-related cues may produce greater motivational salience, and thus are more difficult to extinguish. Using an unbiased place-conditioning paradigm with two doses of cocaine (10 mg/kg or 20 mg/kg), the authors show here that adolescents require 75 +/- 17% more extinction trials than adults to extinguish cocaine place-preferences. Furthermore, once extinguished, adolescents display a greater preference for a previously cocaine-paired environment upon drug-primed reinstatement compared with adults. These results suggest that adolescent vulnerability to addiction involves robust memories for drug-associated cues that are difficult to extinguish. Therefore, drug-addicted adolescents may have a higher risk of relapse than adults, leading to greater prevalence of addiction in this population. PMID- 18410185 TI - Wheel-running behavior is altered following withdrawal from repeated cocaine in adult rats. AB - The residual effects on open-field habituation and self-generated wheel running following withdrawal from repeated cocaine (COC; 30 mg/kg for 7 days) were examined in adult male rats. Control subjects received equivolumetric injections of saline (SAL) and were either allowed to feed ad libitum or pair-fed matched (PF SAL) to COC subjects to control for the drug's potential anorexic effect. Following 10 days of withdrawal, all subjects were examined twice on each of the two assessment instruments. Results indicated that COC subjects over the two test sessions failed to increase their wheel-running rates and did not show the expected habituation in the open field. However, because both COC and PF SAL groups yielded similar effects in the open field, conclusions about cocaine's consequences on habituation could not be established independent of the drug's anorexic effect. These data provide evidence for the view that repeated cocaine impairs motivational processes responsible for engaging in self-generated naturally rewarding behaviors. Speculation concerning the neurobiobehavioral substrates for this effect is presented. PMID- 18410186 TI - Magnitude and timing of nictitating membrane movements during classical conditioning of the rabbit (Oryctolagus cuniculus). AB - A trial-by-trial, subject-by-subject analysis was conducted to determine whether generation of the conditioned response (CR) occurs on a continuous or all-or-none basis. Three groups of rabbits were trained on different partial reinforcement schedules with the conditioned stimulus presented alone on 10%, 30%, or 50%, respectively, of all trials. Plots of each rabbit's nictitating membrane movements revealed that their magnitude rose in a continuous fashion. Response growth during acquisition followed a sigmoidal curve, and the timing of CR-sized movements was largely stable throughout the experiment. The results are discussed with respect to alternative models of CR generation. PMID- 18410187 TI - Impairment in object-in-place scene learning after uncinate fascicle section in macaque monkeys. AB - Three previous experiments have shown that a disconnection of frontal cortex from inferior temporal cortex in monkeys impairs a variety of visual learning tasks but leaves concurrent object discrimination learning intact. In the present experiment, three monkeys were trained on an object-in-place task where concurrent object discrimination learning took place within unique background scenes. After surgery to transect the uncinate fascicle, the monosynaptic route between prefrontal cortex and inferior temporal cortex, all three monkeys showed an impairment relative to their preoperative performance. Combined with previously reported impairments after uncinate fascicle transection, the interaction between frontal cortex and inferotemporal cortex is likely to be important in discrimination learning in background scenes because learning depends on associating the visual elements of a scene together with the appropriate choice object. This result adds to recent evidence showing that tasks such as object-in-place learning and conditional learning are impaired after disconnection of frontal cortex from inferior temporal cortex because those tasks require the representation of temporally extended events. PMID- 18410189 TI - Emotional priming of pop-out in visual search. AB - When searching for a discrepant target along a simple dimension such as color or shape, repetition of the target feature substantially speeds search, an effect known as feature priming of pop-out (V. Maljkovic and K. Nakayama, 1994). The authors present the first report of emotional priming of pop-out. Participants had to detect the face displaying a discrepant expression of emotion in an array of four face photographs. On each trial, the target when present was either a neutral face among emotional faces (angry in Experiment 1 or happy in Experiment 2), or an emotional face among neutral faces. Target detection was faster when the target displayed the same emotion on successive trials. This effect occurred for angry and for happy faces, not for neutral faces. It was completely abolished when faces were inverted instead of upright, suggesting that emotional categories rather than physical feature properties drive emotional priming of pop-out. The implications of the present findings for theoretical accounts of intertrial priming and for the face-in-the-crowd phenomenon are discussed. PMID- 18410188 TI - Social behavior phenotypes in fragile X syndrome, autism, and the Fmr1 knockout mouse: theoretical comment on McNaughton et al. (2008). AB - Comments on the article by C. H. McNaughton et al.. Individuals with fragile X syndrome (FXS) show varying degrees of social behavior disturbances, from social anxiety to autism. This variability of social behavior phenotypes in FXS is likely to be due to interactions of Fmr1 with other gene variants and environmental factors during brain development, although very little is known about the specific genetic and neural mechanisms involved. The Fmr1 knockout mouse is an important experimental resource for elucidating the neural mechanisms of social anxiety, social reward, and social cognition. However, studies of social behavior phenotypes in the Fmr1 knockout mouse are still in early stages. McNaughton et al provide important new information on these phenotypes in the Fmr1 knockout mouse through their use of novel, detailed behavioral analysis to identify signs of increased social anxiety and social cognition deficits. Their significant refinements in measurement of social behavior phenotypes will help to advance future efforts to elucidate the genetic and neural mechanisms underlying social behavior disturbances in FXS and autism. PMID- 18410190 TI - Electrophysiological correlates of encoding and retrieving emotional events. AB - This study examined the impact of emotional content on encoding and retrieval processes. Event-related potentials were recorded in a source recognition memory task. During encoding, a posterior positivity for positive and negative pictures (250-450 ms) that presumably reflects attentional capturing of emotionally valenced stimuli was found. Additionally, positive events, which were also rated as less arousing than negative events, gave rise to anterior and posterior slow wave activity as compared with neutral and negative events and also showed enhanced recognition memory. It is assumed that positive low-arousing events enter controlled and elaborated encoding processes that are beneficial for recognition memory performance. The high arousal of negative events may interfere with controlled encoding mechanisms and attenuate item recognition and the quality of remembering. Moreover, topographically distinct late posterior negativities were obtained for the retrieval of the context features location and time that support the view that this component reflects processes in service of reconstructing the study episode by binding together contextual details with an item and that varies with the kind of episodic detail to be retrieved. PMID- 18410191 TI - Predispositions to approach and avoid are contextually sensitive and goal dependent. AB - The authors show that predispositions to approach and avoid do not consist simply of specific motor patterns but are more abstract functions that produce a desired environmental effect. It has been claimed that evaluating a visual stimulus as positive or negative evokes a specific motor response, extending the arm to negative stimuli, and contracting to positive stimuli. The authors showed that a large congruency effect (participants were faster to approach pleasant and avoid unpleasant stimuli, than to approach unpleasant and avoid pleasant stimuli) could be produced on a novel touchscreen paradigm (Experiment 1), and that the congruency effect could be reversed by spatial (Experiment 2) and nonspatial (Experiment 3) response effects. Thus, involuntary approach and avoid response activations are not fixed, but sensitive to context, and are specifically based on the desired goal. PMID- 18410192 TI - The role of working memory in decoding emotions. AB - Decoding facial expressions of emotion is an important aspect of social communication that is often impaired following psychiatric or neurological illness. However, little is known of the cognitive components involved in perceiving emotional expressions. Three dual task studies explored the role of verbal working memory in decoding emotions. Concurrent working memory load substantially interfered with choosing which emotional label described a facial expression (Experiment 1). A key factor in the magnitude of interference was the number of emotion labels from which to choose (Experiment 2). In contrast the ability to decide that two faces represented the same emotion in a discrimination task was relatively unaffected by concurrent working memory load (Experiment 3). Different methods of assessing emotion perception make substantially different demands on working memory. Implications for clinical disorders which affect both working memory and emotion perception are considered. PMID- 18410193 TI - The impact of changes in spatial distance on emotional responses. AB - Drawing on motivational approaches to emotion, the authors propose that the perceived change in spatial distance to pictures that arouse negative emotions exerts an influence on the significance of these pictures. Two experiments induced the illusion that affective pictures approach toward the observer, recede from the observer, or remain static. To determine the motivational significance of the pictures, emotional valence and arousal ratings as well as startle responses were assessed. Approaching unpleasant pictures were found to exert an influence on both the valence and the arousal elicited by the pictures. Furthermore, movement of pleasant or neutral pictures did not influence startle responses, while the second experiment showed that approaching unpleasant pictures elicited enhanced startle responses compared to receding unpleasant pictures. These findings support the view that a change of spatial distance influences motivational significance and thereby shapes emotional responses. PMID- 18410194 TI - The moderating effects of stimulus valence and arousal on memory suppression. AB - This study examined the separate and combined effects of stimulus valence and arousal on retrieval inhibition. Participants performed Anderson and Green's (2001) memory suppression task with stimuli varying across dimensions of valence and arousal. Memory was tested through free and cued recall as well as speeded recognition. Results showed that both stimulus valence and arousal influenced the extent to which participants successfully inhibited retrieval, but not in the ways anticipated. Specifically, the strongest inhibition effects were for highly arousing, pleasant words. In addition, unpleasant stimuli that were suppressed were better recalled during both cued and free-recall tasks than pleasant stimuli that were suppressed. Across all tests of memory performance, there were no significant differences between the experimental conditions for highly arousing, unpleasant words. The implications of these findings are discussed. PMID- 18410195 TI - The affective regulation of cognitive priming. AB - Semantic and affective priming are classic effects observed in cognitive and social psychology, respectively. The authors discovered that affect regulates such priming effects. In Experiment 1, positive and negative moods were induced before one of three priming tasks; evaluation, categorization, or lexical decision. As predicted, positive affect led to both affective priming (evaluation task) and semantic priming (category and lexical decision tasks). However, negative affect inhibited such effects. In Experiment 2, participants in their natural affective state completed the same priming tasks as in Experiment 1. As expected, affective priming (evaluation task) and category priming (categorization and lexical decision tasks) were observed in such resting affective states. Hence, the authors conclude that negative affect inhibits semantic and affective priming. These results support recent theoretical models, which suggest that positive affect promotes associations among strong and weak concepts, and that negative affect impairs such associations (Clore & Storbeck, 2006; Kuhl, 2000). PMID- 18410196 TI - Facial expression categorization by chimpanzees using standardized stimuli. AB - The ability to recognize and accurately interpret facial expressions are critical social cognition skills in primates, yet very few studies have examined how primates discriminate these social signals and which features are the most salient. Four experiments examined chimpanzee facial expression processing using a set of standardized, prototypical stimuli created using the new ChimpFACS coding system. First, chimpanzees were found to accurately discriminate between these expressions using a computerized matching-to-sample task, and recognition was impaired for all but one expression category when they were inverted. Third, a multidimensional scaling analysis examined the perceived dissimilarity among these facial expressions revealing 2 main dimensions, the degree of mouth closure and extent of lip-puckering and retraction. Finally, subjects were asked to match each facial expression category using only individual component features. For each expression category, at least 1 component movement was more salient or representative of that expression than the others. However, these were not necessarily the only movements implicated in subject's overall pattern of errors. Therefore, similar to humans, both configuration and component movements are important during chimpanzee facial expression processing. PMID- 18410197 TI - Is running away right? The behavioral activation-behavioral inhibition model of anterior asymmetry. AB - The measurement of anterior electroencephalograph (EEG) asymmetries has become an important standard paradigm for the investigation of affective states and traits. Findings in this area are typically interpreted within the motivational direction model, which suggests a lateralization of approach and withdrawal motivational systems to the left and right anterior region, respectively. However, efforts to compare this widely adopted model with an alternative account-which relates the left anterior region to behavioral activation independent of the direction of behavior (approach or withdrawal) and the right anterior region to goal conflict induced behavioral inhibition-are rare and inconclusive. Therefore, the authors measured the EEG in a sample of 93 young men during emotional imagery designed to provide a critical test between the 2 models. The results (e.g., a correlation between left anterior activation and withdrawal motivation) favor the alternative model on the basis of the concepts of behavioral activation and behavioral inhibition. In addition, the present study also supports an association of right parietal activation with physiological arousal and the conceptualization of parietal EEG asymmetry as a mediator of emotion-related physiological arousal. PMID- 18410198 TI - The persistence of attention to emotion: brain potentials during and after picture presentation. AB - Emotional stimuli have been shown to elicit increased perceptual processing and attentional allocation. The late positive potential (LPP) is a sustained P300 like component of the event-related potential that is enhanced after the presentation of pleasant and unpleasant pictures as compared with neutral pictures. In this study, the LPP was measured using dense array electroencephalograph both before and after pleasant, neutral, and unpleasant images to examine the time course of attentional allocation toward emotional stimuli. Results from 17 participants confirmed that the LPP was larger after emotional than neutral images and that this effect persisted for 800 ms after pleasant picture offset and at least 1,000 ms after unpleasant picture offset. The persistence of increased attention after unpleasant compared to pleasant stimuli is consistent with the existence of a negativity bias. Overall, these results indicate that attentional capture of emotion continues well beyond picture presentation and that this can be measured with the LPP. Implications and directions for future research are discussed. PMID- 18410199 TI - Emotion and working memory: evidence for domain-specific processes for affective maintenance. AB - Working memory is comprised of separable subsystems for visual and verbal information, but what if the information is affective? Does the maintenance of affective information rely on the same processes that maintain nonaffective information? The authors address this question using a novel delayed-response task developed to investigate the short-term maintenance of affective memoranda. Using selective interference methods the authors find that a secondary emotion regulation task impaired affect intensity maintenance, whereas secondary cognitive tasks disrupted brightness intensity maintenance, but facilitated affect maintenance. Additionally, performance on the affect maintenance task depends on the valence of the maintained feeling, further supporting the domain specific nature of the task. The importance of affect maintenance per se is further supported by demonstrating that the observed valence effects depend on a memory delay and are not evident with simultaneous presentation of stimuli. These findings suggest that the working memory system may include domain-specific components that are specialized for the maintenance of affective memoranda. PMID- 18410200 TI - Emotion processing effects on interference resolution in working memory. AB - The interaction between emotion and working memory maintenance, load, and performance has been investigated with mixed results. The effect of emotion on specific executive processes such as interference resolution, however, remains relatively unexplored. In this series of studies, we examine how emotion affects interference resolution processes within working memory by modifying the Recency probes paradigm (Monsel, 1978) to include emotional and neutral stimuli. Reaction time differences were compared between interference and non-interference trials for neutral and emotional words (Studies 1 & 3) and pictures (Study 2). Our results indicate that trials using emotional stimuli show a relative decrease in interference compared with trials using neutral stimuli, suggesting facilitation of interference resolution in the former. Furthermore, both valence and arousal seem to interact to produce this facilitation effect. These findings suggest that emotion facilitates response selection amid interference in working memory. PMID- 18410201 TI - A social-cognitive model of trait and state levels of gratitude. AB - Three studies tested a new model of gratitude, which specified the generative mechanisms linking individual differences (trait gratitude) and objective situations with the amount of gratitude people experience after receiving aid (state gratitude). In Study 1, all participants (N = 253) read identical vignettes describing a situation in which they received help. People higher in trait gratitude made more positive beneficial appraisals (seeing the help as more valuable, more costly to provide, and more altruistically intended), which fully mediated the relationship between trait and state levels of gratitude. Study 2 (N = 113) replicated the findings using a daily process study in which participants reported on real events each day for up to 14 days. In Study 3, participants (N = 200) read vignettes experimentally manipulating objective situations to be either high or low in benefit. Benefit appraisals were shown to have a causal effect on state gratitude and to mediate the relationship between different prosocial situations and state gratitude. The 3 studies demonstrate the critical role of benefit appraisals in linking state gratitude with trait gratitude and the objective situation. PMID- 18410202 TI - Moral elevation can induce nursing. AB - There is little extant research on the psychological or physiological response to witnessing good deeds. The authors call the emotional reaction to virtue "moral elevation" and the authors examined its effects on mother-infant dyads. Breastfeeding women who watched a morally elevating video were more likely to nurse their infants and were marginally more likely to hug them, compared to women who watched an equally enjoyable comedy video. Both of these effects suggest that moral elevation may involve the release of oxytocin, a hormone associated with lactation and affiliation. PMID- 18410203 TI - A new mode of fear expression: perceptual bias in height fear. AB - Emotion and psychopathology researchers have described the fear response as consisting of four main components--subjective affect, physiology, cognition, and behavior. The current study provides evidence for an additional component in the domain of height fear (perception) and shows that it is distinct from measures of cognitive processing. Individuals High (N = 35) and Low (N = 36) in acrophobic symptoms looked over a two-story balcony ledge and estimated its vertical extent using a direct height estimation task (visual matching), and an indirect task (size estimation); the latter task seems to exhibit little influence from cognitive factors. In addition, implicit and explicit measures of cognitive processing were obtained. Results indicated that, as expected, the High Fear group showed greater relative, implicit height fear associations and explicit threat cognitions. Of primary interest, the High (compared to Low) Fear group estimated the vertical extent to be higher, and judged target sizes to be greater, even when controlling for the cognitive bias measures. These results suggest that emotional factors such as fear are related to perception. PMID- 18410204 TI - Expressive writing buffers against maladaptive rumination. AB - This study examined whether ruminative style moderated the effects of expressive writing. Sixty-nine participants were assessed for ruminative style and depression symptoms at the beginning of their 1st college semester. Participants were then randomized to either an expressive writing or a control writing condition. Changes in depression symptoms were assessed 2, 4, and 6 months later. Results showed that a brooding ruminative style moderated the effects of expressive writing such that among those assigned to the expressive writing condition, individuals with greater brooding scores reported significantly fewer depression symptoms at all of the follow-up assessments relative to individuals with lower brooding scores. In contrast, reflective pondering ruminative style did not moderate the effects of expressive writing on depression symptoms. These findings suggest that expressive writing could be used as a means of reducing depression symptoms among those with a maladaptive ruminative tendency to brood. PMID- 18410205 TI - The specificity of maternal parenting behavior and child adjustment difficulties: a study of inner-city African American families. AB - The specificity of the association between 2 parenting behaviors (warmth and supervision) and 2 indicators, aggressive behavior and depressive symptoms, of major child outcomes (externalizing problems and internalizing problems) was examined among 196 inner-city African American mothers and their school age children. Given the growing number of African American families affected by HIV/AIDS and demonstrated compromises in parenting associated with maternal infection, the moderating role of maternal HIV/AIDS was also examined. Findings from longitudinal analyses supported the specificity of maternal warmth but not of maternal supervision. Maternal warmth was a stronger predictor of decreases in child aggressive behavior than of decreases in depressive symptoms. In addition, maternal warmth was a stronger predictor of decreases in aggressive behavior than was maternal supervision. Parenting specificity was not moderated by maternal HIV/AIDS. Clinical implications and future research directions are discussed. PMID- 18410206 TI - Evaluating a brief prevention program for improving marital conflict in community families. AB - Marital conflict is related to well-being in children and adults (E. M. Cummings & P. T. Davies, 2002). Marital conflict is likely most effectively ameliorated before it becomes clinically significant. However, families without significant problems may be unwilling to participate in couples therapies or other lengthy or intensive interventions. Responding to this gap, the authors developed a 4 session psychoeducational program about marital conflict for community families. Couples with children 4-8 years of age were randomly blocked into 1 of 3 groups: (1) a parent-only group (n = 24), (2) a parent-child group (n = 33), or (3) a self-study group (n = 33). Pre- and posttest and 6-month and 1-year assessments were conducted. This report evaluates (a) whether participation in a psychoeducational program for parents improved marital conflict, especially concerning ways of expressing disagreements, and (b) whether changes in marital conflict subsequently improved marital satisfaction, parenting, and child adjustment. Greater constructive and less destructive marital conflict was observed at all assessments for treatment groups, and these changes were linked with improvements in other family processes. The findings support the promise of brief, psychoeducational programs for improving marital conflict for community samples. PMID- 18410207 TI - Long-term effects of the death of a child on parents' adjustment in midlife. AB - The death of a child is a traumatic event that can have long-term effects on the lives of parents. This study examined bereaved parents of deceased children (infancy to age 34) and comparison parents with similar backgrounds (n = 428 per group) identified in the Wisconsin Longitudinal Study. An average of 18.05 years following the death, when parents were age 53, bereaved parents reported more depressive symptoms, poorer well-being, and more health problems and were more likely to have experienced a depressive episode and marital disruption than were comparison parents. Recovery from grief was associated with having a sense of life purpose and having additional children but was unrelated to the cause of death or the amount of time since the death. The results point to the need for detection and intervention to help those parents who are experiencing lasting grief. PMID- 18410208 TI - Sense of control predicts depressive and anxious symptoms across the transition to parenthood. AB - In this study, the authors examined the relationship between sense of control and depressive and anxious symptoms for mothers and fathers during the 1st year of parenthood. Participants were 153 dual-earner, working-class couples who were recruited during the 3rd trimester of pregnancy at prenatal education courses. Data were collected 1 month antenatally and 1, 4, 6, and 12 months postnatally. Sense of control was decomposed into 2 distinct parts: an enduring component and a malleable component that changes with context. Consistent with a cognitive theory of emotional problems, results demonstrated that a sense of control served a protective function for mental health outcomes. A higher sense of enduring control predicted lower levels of psychological distress for new parents, and increases in control over time predicted decreases in depression and anxiety. Findings hold implications for interventions with expectant parents, such as expanding prenatal education courses to include strategies for enhancing and maintaining a sense of personal control. PMID- 18410209 TI - Ambiguous absence, ambiguous presence: a qualitative study of military reserve families in wartime. AB - The "Global War on Terrorism" has resulted in reservists being deployed at an ever-increasing rate. However, because reservists and their families are unaccustomed to deployments, many families may experience boundary ambiguity, a state in which family members are uncertain in their perception about who is in or out of the family and who is performing which roles and tasks within the family. This qualitative description study examined boundary ambiguity in military reserve families over time. A sample of 34 reservists, spouses, and parents was interviewed 7 times within the 1st year of the reservists' return from Iraq. During deployment, all family members experienced boundary ambiguity. Gathering information and attending a family support group provided some relief for families. After the reservists returned, couples as well as those who had experienced additional life events or losses experienced the highest levels of boundary ambiguity. However, this boundary ambiguity dissipated over time, as families tended to restabilize once the reservists had returned to work and a routine had been established. PMID- 18410210 TI - Longitudinal associations between parents' hostility and siblings' externalizing behavior in the context of marital discord. AB - This study examined longitudinal associations between parents' hostility and siblings' externalizing behavior in the context of interparental discord. The sample included 116 families (mothers, fathers, 2 siblings) assessed in middle childhood, when siblings were, on average, 8 and 10 years old, and in adolescence, at average ages of 14 and 16 years. Parents reported on their hostility toward each child and on each child's externalizing problems. Raters observed interparental hostility, and parents rated their marital quality. Results indicated both within-family and between-families effects. Specifically, the child who received more parental hostility than his or her sibling showed greater increases in externalizing problems than his or her sibling; this association was moderated by marital discord. In addition, the child who exhibited more behavioral problems than his or her sibling received greater increases in hostile mothering than did his or her sibling. Between-families effects were evident, in that children's externalizing problems were associated with increases in mothers' hostility toward both children in the family. Results support transactional models of development and family systems theory. PMID- 18410211 TI - Change in caregiver depression as a function of the Strong African American Families Program. AB - A randomized prevention trial contrasted families who participated in the Strong African American Families Program (SAAF), a preventive intervention for rural African American parents and their 11-year-olds, with control families. This article focuses on the program's effect on primary caregivers' depressive symptoms. Among the 167 caregivers with elevated scores on the Center for Epidemiologic Studies-Depression Scale, SAAF participation was associated with reduced depressive symptoms, enhanced parenting, and perceived improvements in youth behavior. Change in parenting (consistent discipline, youth monitoring, and open communication) but not change in youth intrapersonal competencies significantly mediated intervention effects on caregivers' depression. Results support the link between reduced depressive symptoms and stronger family relationships, particularly the importance of enhanced parenting efficacy in alleviating depressive symptoms. PMID- 18410213 TI - Maternal depression and parenting in relation to child internalizing symptoms and asthma disease activity. AB - This study tested a hypothesized model of the relationship between maternal depression and child psychological and physical dysfunction mediated by parenting and medication adherence. A sample of 242 children with asthma, aged 7 to 17, participated with their mothers. Maternal depression was assessed by self-report, and parenting was observed during family interaction tasks. Internalizing symptoms were assessed by self- and clinician reports. Asthma disease activity was assessed according to National Heart Lung and Blood Institute guidelines, and medication adherence was evaluated with a 24-hr recall method. Structural equation modeling indicated that negative parenting partially mediated the relationship between maternal depression and child internalizing symptoms. Child internalizing symptoms, in turn, mediated the associations between both maternal depression and negative parenting and asthma disease activity. Medication adherence did not mediate the link from maternal depression to disease activity. Thus, maternal depression was linked to child psychological dysfunction both directly and indirectly via negative parenting but linked to physical dysfunction only indirectly through psychological dysfunction. These findings suggest that diagnosing and treating depression in mothers of children with asthma would enhance child well-being both psychologically and physically. PMID- 18410212 TI - Establishing family foundations: intervention effects on coparenting, parent/infant well-being, and parent-child relations. AB - This study investigated the ability of a theoretically driven, psychosocial prevention program implemented through childbirth education programs to enhance the coparental relationship, parental mental health, the parent-child relationship, and infant emotional and physiological regulation. A sample of 169 heterosexual, adult couples who were expecting their 1st child was randomized to intervention and control conditions. The intervention families participated in Family Foundations, a series of 8 classes, delivered before and after birth, that was designed as a universal prevention program (i.e., it was applicable to all couples, not just those at high risk). Intent-to-treat analyses indicated significant program effects on coparental support, maternal depression and anxiety, distress in the parent-child relationship, and several indicators of infant regulation. Intervention effects were not moderated by income, but greater positive impact of the program was found for lower educated parents and for families with a father who reported higher levels of insecure attachment in close relationships. These findings support the view that coparenting is a potentially malleable intervention target that may influence family relationships as well as parent and child well-being. PMID- 18410214 TI - Prospective associations from family-of-origin interactions to adult marital interactions and relationship adjustment. AB - To test the social learning-based hypothesis that marital conflict resolution patterns are learned in the family of origin, longitudinal, observational data were used to assess prospective associations between family conflict interaction patterns during adolescence and offspring's later marital conflict interaction patterns. At age 14 years, 47 participants completed an observed family conflict resolution task with their parents. In a subsequent assessment 17 years later, the participants completed measures of marital adjustment and an observed marital conflict interaction task with their spouse. As predicted, levels of hostility and positive engagement expressed by parents and adolescents during family interactions were prospectively linked with levels of hostility and positive engagement expressed by offspring and their spouses during marital interactions. Family-of-origin hostility was a particularly robust predictor of marital interaction behaviors; it predicted later marital hostility and negatively predicted positive engagement, controlling for psychopathology and family-of origin positive engagement. For men, family-of-origin hostility also predicted poorer marital adjustment, an effect that was mediated through hostility in marital interactions. These findings suggest a long-lasting influence of family communication patterns, particularly hostility, on offspring's intimate communication and relationship functioning. PMID- 18410215 TI - Risky sexual behavior among married alcoholic men. AB - The current study explored whether the wives of men entering alcoholism treatment are at risk for sexually transmitted infections (STIs) exposure as a result of their husbands' sexual risk behaviors. The extramarital relationships of married alcoholic men entering outpatient treatment (n = 125) were compared with those of a demographically matched community sample of nonalcoholic married men (n = 125). The proportion of alcoholic men who reported 1 or more extramarital affairs in the previous year (14%) was significantly higher than that of the community sample (4%). Additionally, only 2 alcoholic husbands and 1 nonalcoholic husband reported that his wife was aware of the extramarital relationship. For both groups, none of the men who engaged in extramarital relationships reported consistent use of condoms when having sexual intercourse with their wives or with their extramarital partners. These results suggest that wives of alcoholic men are unknowingly placed at risk for indirect exposure to STIs as a result of their husbands' sexual risk behaviors. Thus, infidelity in treatment-seeking alcohol abusing men represents a significant public health issue. PMID- 18410217 TI - Intention to communicate BRCA1/BRCA2 genetic test results to the family. AB - Guided by the theory of planned behavior, this analysis explores the communication skills of women who had genetic testing for BRCA1 and BRCA2. The key outcome was intention to tell test results to adult first-degree relatives. The theory predicts that global and specific attitudes, global and specific perceived social norms, and perceived control will influence the communication of genetic test results. A logistic regression model revealed that global attitude (p < .05), specific social influence (p < .01), and perceived control (p < .05) were significant predictors of intention to tell. When gender and generation of relatives were added to the regression, participants were more likely to convey genetic test results to female than to male relatives (p < .05) and were also more likely to communicate test results to children (p < .01) or siblings (p < .05) than to parents. However, this association depended on knowing the relative's opinion of genetic testing. Intention to tell was lowest among participants who did not know their relative's opinion. These results extend the theory of planned behavior by showing that gender and generation influence intention when the relative's opinion is unknown. PMID- 18410216 TI - Sampling and recruitment in studies of cultural influences on adjustment: a case study with Mexican Americans. AB - Research examining how cultural factors affect adjustment of ethnic minority individuals would be strengthened if study samples better represented the diversity within these populations. To recruit a representative sample of Mexican American families, the authors implemented a multiple-step process that included sampling communities to represent diversity in cultural and economic conditions, recruiting participants through schools, using culturally attractive recruitment processes, conducting interviews in participants' homes, and providing a financial incentive. The result was a sample of 750 families that were diverse in cultural orientation, social class, and type of residential communities and were similar to the census description of this population. Thus, using culturally appropriate adaptations to common recruitment strategies makes it possible to recruit representative samples of Mexican Americans. PMID- 18410218 TI - Attachment insecurity and perceptions of housework: Associations with marital well-being. AB - This exploratory study examines the interactive effects of attachment insecurity and perceptions of housework on 2 dimensions of marital well-being--satisfaction and perceptions of fairness. Participants were 148 married couples obtained from an area probability sample as part of a larger study. Multilevel modeling analyses with the couple as the unit of analysis showed that women who scored high and men who scored low on the dimension of attachment anxiety and reported that their spouses performed more routine housework (i.e., prepares meals) also reported being over-benefited. Women who scored high and men who scored low on the dimension of attachment avoidance and reported that their spouses performed more intermittent housework (i.e., yard work) reported greater marital satisfaction. These results highlight the role of attachment orientations in explaining why perceptions of housework may have more or less prominent effects on marital well-being. PMID- 18410219 TI - Child sexual abuse, early family risk, and childhood parentification: pathways to current psychosocial adjustment. AB - In this study, the authors examined the role of parentification (children assuming adult-like roles in the family) as it relates to family risk (parental psychopathology, parental illness, and domestic violence), child sexual abuse (CSA), and psychosocial adjustment in 499 college women. Structural equation modeling was used to test a model of direct, indirect, and mediational pathways through which CSA, family risk, and parentification contributed to later psychosocial maladjustment. Results indicate that CSA and family risk independently and directly predicted higher levels of maladjustment, but only family risk positively predicted parentification in childhood. Parentification was unexpectedly related to less maladjustment. Parentification failed to mediate the relation between early family risk and maladjustment. Findings suggest that family risk factors may contribute to parentification and that parentification is not always related to poorer psychosocial outcomes. Future research should examine the impact of parentification on other aspects of functioning and should assess how individual, familial, and cultural variables (e.g., age, gender, duration, perceived fairness, ethnicity, and family support) moderate the impact of parentification on long-term adjustment. PMID- 18410220 TI - Family functioning in families of children with anxiety disorders. AB - The authors examined maternal and paternal reports of family functioning and their relationship with child outcomes as well as the association between anxiety and depression in family members and family functioning. Results reveal that maternal and paternal reports of family functioning were both significantly associated with worse child outcomes, including child anxiety disorder (AD) severity, anxiety symptoms, and child global functioning. Maternal and paternal anxiety and depression predicted worse family functioning, whereas child report of anxiety and depression did not. Parents of children with ADs reported significantly worse family functioning and behavior control, but only fathers reported worse problem solving and affective involvement compared with fathers of children with no psychological disorders. Findings from this study suggest that paternal as well as maternal anxiety and depression play a role in worse family functioning in children with ADs and that unhealthier family functioning is associated with worse child outcomes in this population. PMID- 18410221 TI - Forty years of allocated seats for Sami medical students - has preferential admission worked? AB - INTRODUCTION: This article examines the effects of a special admission policy for Sami medical students in Norway. In the 1960s, public health and health care were found to be poorer in Sami communities than in the rest of Norway. There were few doctors and none of them spoke Sami. Sami school leavers found it difficult to gain admittance to medical schools. In response to this situation, the medical faculty at the University of Bergen adopted a special admissions policy for Sami students in 1963. The University of Tromso did the same in 1991. In this study we have analyzed whether the allocated Sami seats produced the desired outcomes. In assessing the outcomes, the study takes into account the considerable improvements in public health and health care in the last 40 years, wider use of the Sami language and generally higher educational achievements among the Sami. METHODS: This retrospective study was set in two medical schools in Norway. The study population is students admitted to medical school on allocated Sami seats, in the two periods 1963-1986 at the University of Bergen, and 1991-2000 at the University of Tromso. After a question identified the Sami students, whether they had practised or were practising medicine was determined. RESULTS: In total 38 students were admitted on the allocated Sami seats, and 32 graduated. Of the candidates, 93% had practised medicine in one of the two northernmost counties in Norway. Graduates during the 1960s and 1970s were more likely to have worked as GPs in the main areas of Sami habitation than the Sami physicians who graduated later. CONCLUSIONS: The Sami doctors admitted to medical school on allocated Sami seats have practiced in Finnmark or Troms, counties where most of the Sami people live. However, this study was unable to establish whether admission on these grounds led to more Sami doctors working in the main areas of Sami habitation. Regarding the workplace location variable, there were no differences between Sami and other physicians from the northern part of Norway who were educated at the University of Tromso. PMID- 18410222 TI - Quality of life in patients with sickle cell disease in Jamaica: rural-urban differences. AB - INTRODUCTION: Quality of life (QOL) refers to people's ability to function in the ordinary tasks of living. It moves beyond direct manifestations of illness to the patient's personal morbidity. These assessments are an important aspect of chronic disease management. Sickle cell disease (SCD) is a chronic and potentially, quite a debilitating disease. The disease is severe and may result in significant morbidity, as well as a shortened life span. It is the most common genetic disorder seen in Jamaica and impacts on physical, psychological, social and occupational wellbeing. Jamaica is a developing country where support systems that exist for patients with SCD are sparse. Health related QOL has been shown to be poorer in people living in the rural areas as compared with urban populations. Utilization of comprehensive sickle cells disease services has also been shown to be lower for individuals with the disease living in rural areas than for those living in urban areas. As there are rural-urban differences in Jamaica's health services, it is hypothesized that there may be rural-urban differences in the experiences of the disease and the QOL of these patients in these subgroups. METHODS: The SF 36 v2 (Short Form 36) questionnaire has been validated for use in the Jamaican SCD population. This validated questionnaire was interviewer administered to 166 patients presenting to an urban clinic for routine health maintenance visits and to 90 patients presenting to the rural clinics for routine visits. Socio-demographic information was also collected on these two groups. Multiple linear regression analyses were performed to study predictors of QOL in these two sub-populations. The study received ethical approval from the University of the West Indies/University Hospital of the West Indies Ethics Committee. RESULTS: There were no significant differences in the measured socio demographic characteristics of the rural and urban patients. Living in rural areas compared with urban areas (p <0.001), being employed (p 0.008), and having the heterozygous SC disease versus homozygous sickle cell (SS) disease (p 0.04) were all associated with improved QOL. The rural group also had better physical and mental health scores, as well as perceiving fewer limitations in their daily living activities as a result of their disease. CONCLUSIONS: The study shows that the rural SCD sample assessed their QOL as higher than the urban sample. It is possible that this surprising finding may be the result of lower levels of social support for urban SCD patients. However, further work could help elucidate why rural subgroups of people with this disease appear to enjoy much better QOL despite limited health care access. PMID- 18410224 TI - Berberine promotes osteoblast differentiation by Runx2 activation with p38 MAPK. AB - Berberine (BBR) has been implicated in bone biology. Although BBR reduces osteoporosis by enhancing BMD and inhibiting osteoclast activity, the effects of BBR on osteoblasts during the process of osteogenesis have not been thoroughly studied. In osteoblastic cells, BBR enhanced the expression of osteogenic marker genes including osteopontin and osteocalcin and promoted the transcriptional activity of the key osteogenic transcription factor Runx2. In osteoblasts, BBR increased the binding of Runx2 to the promoter region of osteopontin. The recruitment of co-factors such as p300 and HDAC1 to the promoter regions of osteopontin and osteocalcin was regulated by BBR, resulting in an enhancement in the expression of those genes. Furthermore, BBR activated p38 mitogen-activated protein kinase (MAPK) and increased cyclooxygenase 2 (COX2) expression, which are key factors in osteoblast differentiation. Consistently, a p38 MAPK-specific inhibitor attenuated the effect of BBR on osteogenesis, whereas p38 MAPK overexpression augmented BBR-induced osteogenic gene expression. Moreover, BBR stimulated bone area formation in calvarial organ culture. Taken together, these findings indicate that BBR promotes osteoblast differentiation through activation of Runx2 by p38 MAPK. Therefore, BBR may be a potential therapeutic agent to treat bone-related disorders including osteoporosis. PMID- 18410225 TI - Randomized controlled trial of the effects of calcium with or without vitamin D on bone structure and bone-related chemistry in elderly women with vitamin D insufficiency. AB - There are few data on the relative effects of calcium supplementation with or without extra vitamin D on BMD in patients selected for low vitamin D status. The aim of this study is to evaluate the relative importance of vitamin D and calcium treatment on BMD and bone-related chemistry in elderly women with vitamin D insufficiency. Three hundred two elderly women (age, 77.2 +/- 4.6 yr) with serum 25(OH)D concentrations <60 nM participated in a 1-yr randomized, double-blind, placebo-controlled trial. All subjects received 1000 mg calcium citrate per day with either 1000 IU ergocalciferol (vitamin D(2)) or identical placebo (control). The effects of time and time treatment interactions were evaluated by repeated measures ANOVA. At baseline, calcium intake was 1100 mg/d, and 25(OH)D was 44.3 +/- 12.9 nM; this increased in the vitamin D group by 34% but not the control group after 1 year (59.8 +/- 13.8 versus 45.0 +/- 13.3 nM, p < 0.001). Total hip and total body BMD increased significantly, and procollagen type I intact N terminal propeptide (PINP) decreased during the study with no difference between the treatment groups (hip BMD change: vitamin D, +0.5%; control, +0.2%; total body BMD change: vitamin D, +0.4%; control, +0.4%; PINP change: vitamin D, -3.9%; placebo, -2.8%). Although the fasting plasma and urine calcium increased in both groups equally, there was no detectable change in serum PTH. The increase in 25(OH)D achieved with vitamin D supplementation had no extra effect on active fractional intestinal calcium absorption, which fell equally in both groups (vitamin D, -17.4%; control, -14.8%). In patients with a baseline calcium intake of 1100 mg/d and vitamin D insufficiency, vitamin D(2) 1000 IU for 1 year has no extra beneficial effect on bone structure, bone formation markers, or intestinal calcium absorption over an additional 1000 mg of calcium. Vitamin D supplementation adds no extra short-term skeletal benefit to calcium citrate supplementation even in women with vitamin D insufficiency. PMID- 18410226 TI - Significance of serum TRACP in rheumatoid arthritis. AB - Human serum contains two related isoforms of TRACP: TRACP 5a and TRACP 5b. Serum TRACP 5a protein is increased in about one third of rheumatoid arthritis (RA) sera. This study was undertaken to examine the significance of serum TRACP isoforms 5a and 5b as disease markers of inflammation and bone destruction in RA. One hundred eighteen patients were recruited including 50 with RA (25 with nodules), 26 with osteoarthritis (OA), and 42 with other rheumatic diseases. Twenty-six healthy adults served as controls. Serum TRACP 5a activity, TRACP 5a protein, and TRACP 5b activity were determined by in-house immunoassays. C reactive protein (CRP) was determined by in-house immunoassay using commercial antibodies and CRP. Other commercial markers included bone-specific alkaline phosphatase (BALP), C-telopeptides of type-I collagen (ICTP), cartilage glycoprotein-39 (YKL-40), and IgM rheumatoid factors (IgM-RF). Mean TRACP 5a protein was significantly elevated only in RA compared with healthy controls and other disease groups. TRACP 5a protein correlated significantly only with IgM-RF in RA. Among RA patients, mean TRACP 5a protein and IgM RF were significantly higher in nodule formers. In contrast, TRACP 5b activity was slightly elevated in RA and correlated with BALP, ICTP, and YKL-40 but not with IgM-RF or CRP. Mean TRACP 5b activity was no different in RA patients with or without nodules. TRACP isoforms could be useful disease markers in RA; TRACP 5a protein may be a measure of systemic inflammatory macrophage burden and disease severity. TRACP 5b activity is a marker for osteoclast number and perhaps local or systemic bone destruction. PMID- 18410227 TI - Low serum 25-hydroxyvitamin D levels associated with falls among Japanese community-dwelling elderly. AB - Previous studies have shown that low serum 25-hydroxyvitamin D [25(OH)D] level is a risk factor for falls among the elderly in European and North American populations. We used a cross-sectional community-based survey to study the association of serum 25(OH)D level and falls among Japanese community-dwelling elderly. A total of 2957 elderly persons (950 men and 2007 women) 65-92 yr of age who participated in mass health examinations for the prevention of geriatric syndrome for the elderly underwent an interview, blood analysis, and physical performance testing. Experience of falls over the previous year was assessed in an interview. Physical performance tests of handgrip strength, stork standing time with the eyes open, and normal waking speed as risk factors for falls among the elderly were conducted. Serum albumin and 25(OH)D concentrations were analyzed. Mean 25(OH)D concentration was significantly lower in women than in men (p < 0.001). Women showed a significant decline of 25(OH)D level with increased age (p < 0.001). There was also a significant difference in the prevalence of 25(OH)D insufficiency [25(OH)D level < 20 ng/ml] between the sexes (p < 0.001). The rate of falls was significantly higher in the lowest quartile of 25(OH)D level in women (p = 0.02) and in women with 25(OH)D insufficiency (p = 0.001). Women also showed significant declines in all three fall-related physical performance tests. Multiple logistic regression analysis showed significant and independent associations between 25(OH)D level and experience of falls in women only (p = 0.01). Low 25(OH)D level was significantly associated with a high prevalence of falls in Japanese elderly women because of their inferior physical performance. Low serum 25(OH)D levels appear preventable and easily treated; there is an evident need for greater awareness to screen and thus prevent this condition. PMID- 18410228 TI - Vitamin D receptor-dependent 1 alpha,25(OH)2 vitamin D3-induced anti-apoptotic PI3K/AKT signaling in osteoblasts. AB - Osteoblast apoptosis plays a crucial role in bone remodeling. Physiological doses of 1 alpha,25(OH)(2)-vitamin D(3) (1,25D) protect osteoblasts against apoptosis by means of mechanisms only partially understood. We studied activation of an Akt survival cascade downstream of 1,25D nongenomic stimulation of phosphatidylinositide-3'-kinase (PI3K) in osteoblastic cells. We measured a dose- and time-dependent 1,25D induction of Akt phosphorylation (p-Akt) in cultured osteoblastic cells. Maximal response was achieved with 10 nM 1,25D after 5 min. We found that staurosporine (STSP)-induced apoptosis was significantly reduced in 1,25D-pretreated osteoblasts. 1,25D prosurvival effects were abolished when cells were preincubated with inhibitors of PI3K activation. By means of siRNA silencing, we proved that 1,25D induction of p-Akt requires a classic vitamin D receptor (VDR) in osteoblasts. Furthermore, non-osteoblastic CV-1 cells transfected with an enhanced green fluorescent protein (EGFP)-VDR construct responded to 1,25D treatment with a rapid p-Akt response associated with increased cell survival not detected in native, nontransfected cells. We measured increased levels of p-Akt substrates p-Bad and p-FKHR and significantly reduced activity of caspases 8 and 3/7 after 1,25D treatment. In addition, 1,25D-induced protection against apoptosis was abolished when osteoblasts were preincubated with pertussis toxin. We conclude that anti-apoptotic effects of 1,25D in osteoblasts occur through nongenomic activation of a VDR/PI3K/Akt survival pathway that includes phosphorylation of multiple p-Akt substrates and reduction of caspase activities. PMID- 18410229 TI - BMD and bone geometry in transtibial and transfemoral amputees. AB - Prolonged unloading of bone(s) in the residual limb after amputation may cause significant bone loss in the hip and distal bony end of the residual limb. The purpose of this study was to examine the effect of amputation on bone geometry, volumetric BMD (vBMD), and areal BMD (aBMD) by comparing the intact and residual limbs in unilateral transfemoral and transtibial amputees. Amputees (seven above knee; seven below-knee) and two groups of nonamputee control subjects gave informed consent to participate in this study. aBMD of the dual proximal femur, lumbar spine, and total body was assessed using DXA. Bone geometry and vBMD were assessed at the distal ends of the residual limb and intact limb and at a comparable cross-sectional slice of the intact limb using pQCT (Stratec XCT 3000). There were no significant group differences in age, height, weight, physical activity, time as an amputee, hours wearing a prosthesis per day, or total body and lumbar spine BMD and BMC. There were significant side x group interactions for total hip, femoral neck, and trochanter BMD, with the amputated side having lower BMD, and differences being most severe in above-knee amputees. Total and cortical vBMD and area were significantly lower at the end of the residual limb compared with the similar slice of the intact limb for both above- and below-knee amputees. In conclusion, amputees exhibited large decrements in BMD, both at the hip and at the end of the residual limb, compared with the intact side. These lower BMD values put amputees, particularly the above-knee amputees, at increased risk for osteoporosis and fragility fractures in the hip. PMID- 18410231 TI - Sodium and bone health: impact of moderately high and low salt intakes on calcium metabolism in postmenopausal women. AB - High salt intake is a well-recognized risk factor for osteoporosis because it induces calciuria, but the effects of salt on calcium metabolism and the potential impact on bone health in postmenopausal women have not been fully characterized. This study investigated adaptive mechanisms in response to changes in salt and calcium intake in postmenopausal women. Eleven women completed a randomized cross-over trial consisting of four successive 5-wk periods of controlled dietary intervention, each separated by a minimum 4-wk washout. Moderately low and high calcium (518 versus 1284 mg) and salt (3.9 versus 11.2 g) diets, reflecting lower and upper intakes in postmenopausal women consuming a Western-style diet, were provided. Stable isotope labeling techniques were used to measure calcium absorption and excretion, compartmental modeling was undertaken to estimate bone calcium balance, and biomarkers of bone formation and resorption were measured in blood and urine. Moderately high salt intake (11.2 g/d) elicited a significant increase in urinary calcium excretion (p = 0.0008) and significantly affected bone calcium balance with the high calcium diet (p = 0.024). Efficiency of calcium absorption was higher after a period of moderately low calcium intake (p < 0.05) but was unaffected by salt intake. Salt was responsible for a significant change in bone calcium balance, from positive to negative, when consumed as part of a high calcium diet, but with a low calcium intake, the bone calcium balance was negative on both high and low salt diets. PMID- 18410230 TI - Relationship of blood lead levels to incident nonspine fractures and falls in older women: the study of osteoporotic fractures. AB - Lead is stored in the skeleton and can serve as an endogenous source for many years. Lead may influence the risk of fracture, through direct effects on bone strength or indirectly by disturbing neuromuscular function and increasing the risk of falls. The objective of this analysis is to test the hypothesis that women with higher blood lead levels experience higher rates of falls and fracture. This was a prospective cohort study of 533 women 65-87 yr of age enrolled in the Study of Osteoporotic Fractures at two U.S. research centers (Baltimore, MD; Monongahela Valley, PA) from 1986 to 1988. Blood lead levels (in microg/dl) were measured in 1990-1991 by atomic absorption spectrophotometry and classified as "low" (or=8; upper 15th percentile). Total hip BMD was measured by DXA twice, 3.55 yr apart. Information on falls was collected every 4 mo for 4 yr. Incident nonspine fractures were identified and confirmed over 10 yr. Cox proportional hazards models were used to estimate the hazard ratio (HR) and 95% CI of fracture. Generalized estimating equations were used to calculate the incident rate ratio of falls (95% CI). The mean blood lead level was 5.3 +/- 2.3 (SD) microg/dl (range, 1-21 microg/dl). Baseline BMD was 7% lower in total hip and 5% lower in femoral neck in the highest compared with lowest blood lead group (p < 0.02). Hip bone loss tended to be greater in the high lead group, but differences were not significant. In multivariable adjusted models, women with high blood lead levels had an increased risk of nonspine fracture (HR = 2.50; 95% CI = 1.25, 5.03; p trend = 0.016) and higher risk of falls (incident rate ratio = 1.62; 95% CI = 1.07, 2.45; p trend = 0.014) compared with women with lowest lead level. Blood lead levels are associated with an increased risk of falls and fractures, extending the negative health consequences of lead to include osteoporotic fractures. PMID- 18410232 TI - Theoretical implications of the biomechanical fracture threshold. AB - Because of the dichotomous nature of a bone fracture, when Phi, the ratio of the applied impact force to the bone strength, is greater than a critical value--the biomechanical fracture threshold--fracture should occur. We sought to elucidate the conceptual implications of this biomechanical fracture threshold with application to hip fracture. We used data from the PaTH study, a 2-yr clinical trial in postmenopausal women treated with alendronate, PTH, or their combination. Outcomes included the force applied to the hip in a sideways fall as estimated from subject height and weight; femoral strength as determined by QCT based finite element analysis; the load-to-strength ratio Phi; and total hip areal BMD from DXA. Results indicated that those with "very low" femoral strength (<2000 N) invariably had load-to-strength ratio Phi values well above the theoretical biomechanical fracture threshold (Phi = 1), but those with "moderately low" femoral strength (2000-4000 N) displayed Phi values both above and below the theoretical biomechanical fracture threshold. This finding implies that the risk of a hip fracture can be high in those with only moderately low BMD because femoral strength can be low relative to fall impact forces. The observed weak correlation between areal BMD and the load-to-strength ratio Phi (r2 = 0.14) suggests that consideration of the biomechanical fracture threshold may improve fracture risk assessment, particularly for those in the osteopenic range. Regarding treatment effects, only those subjects having load-to-strength ratio Phi values within a relatively narrow "transition zone" of +/- 20% of the assumed biomechanical fracture threshold at baseline were predicted to change fracture status during the trial. In theory, outcomes of fracture trials may be dominated by the responses of those within the "transition zone" at baseline, and treatment benefits in terms of fracture efficacy may depend the patient's baseline status with respect to the biomechanical fracture threshold. We conclude that consideration of the theoretical implications of the biomechanical fracture threshold may lead to new insights and advances in the assessment and treatment of osteoporosis. PMID- 18410233 TI - Functional adaptation to loading of a single bone is neuronally regulated and involves multiple bones. AB - Regulation of load-induced bone formation is considered a local phenomenon controlled by osteocytes, although it has also been hypothesized that functional adaptation may be neuronally regulated. The aim of this study was to examine bone formation in multiple bones, in response to loading of a single bone, and to determine whether adaptation may be neuronally regulated. Load-induced responses in the left and right ulnas and humeri were determined after loading of the right ulna in male Sprague-Dawley rats (69 +/- 16 days of age). After a single period of loading at -760-, -2000-, or -3750-microepsilon initial peak strain, rats were given calcein to label new bone formation. Bone formation and bone neuropeptide concentrations were determined at 10 days. In one group, temporary neuronal blocking was achieved by perineural anesthesia of the brachial plexus with bupivicaine during loading. We found right ulna loading induces adaptive responses in other bones in both thoracic limbs compared with Sham controls and that neuronal blocking during loading abrogated bone formation in the loaded ulna and other thoracic limb bones. Skeletal adaptation was more evident in distal long bones compared with proximal long bones. We also found that the single period of loading modulated bone neuropeptide concentrations persistently for 10 days. We conclude that functional adaptation to loading of a single bone in young rapidly growing rats is neuronally regulated and involves multiple bones. Persistent changes in bone neuropeptide concentrations after a single loading period suggest that plasticity exists in the innervation of bone. PMID- 18410234 TI - In vivo microMRI-based finite element and morphological analyses of tibial trabecular bone in eugonadal and hypogonadal men before and after testosterone treatment. AB - Osteoporosis is a major public health problem in men. Hypogonadal men have decreased BMD and deteriorated trabecular bone architecture compared with eugonadal men. Testosterone treatment improves their BMD and trabecular structure. We tested the hypothesis that testosterone replacement in hypogonadal men would also improve their bone's mechanical properties. Ten untreated severely hypogonadal and 10 eugonadal men were selected. The hypogonadal men were treated with a testosterone gel for 24 mo to maintain their serum testosterone concentrations within the normal range. Each subject was assessed before and after 6, 12, and 24 mo of testosterone treatment by microMRI of the distal tibia. A subvolume of each microMR image was converted to a microfinite element (microFE) model, and six analyses were performed, representing three compression and three shear tests. The anisotropic stiffness tensor was calculated, from which the orthotropic elastic material constants were derived. Changes in microarchitecture were also quantified using newly developed individual trabeculae segmentation (ITS)-based and standard morphological analyses. The accuracy of these techniques was examined with simulated microMR images. Significant differences in four estimated anisotropic elastic material constants and most morphological parameters were detected between the eugonadal and hypogonadal men. No significant change in estimated elastic moduli and morphological parameters was detected in the eugonadal group over 24 mo. After 24 mo of treatment, significant increases in estimated elastic moduli E(22) (9.0%), E(33) (5.1%), G(23) (7.2%), and G(12) (9.4%) of hypogonadal men were detected. These increases were accompanied by significant increases in trabecular plate thickness. These results suggest that 24 mo of testosterone treatment of hypogonadal men improves estimated elastic moduli of tibial trabecular bone by increased trabecular plate thickness. PMID- 18410235 TI - Expression and regulation of the vitamin D receptor in the zebrafish, Danio rerio. AB - Vitamin D and vitamin D metabolites such as 25-hydroxyvitamin D and 1alpha,25 dihydroxyvitamin D [1alpha,25(OH)(2)D(3)] circulate in the serum of fish. The receptor for 1alpha,25(OH)(2)D(3) (VDR) has previously been cloned from fish intestine, and ligand binding assays have shown the presence of the VDR in the gills, intestine, and liver of fish. Using immunohistochemical methods with specific antibodies against the VDR, we now report that the VDR is widely expressed in tissues of the adult male and female zebrafish, Danio rerio, specifically in epithelial cells of gills, tubular cells of the kidney, and absorptive cells in the intestine. Additionally, the VDR is expressed in the skin, the olfactory organ, the retina, brain, and spinal cord. Sertoli cells of the testis, oocytes, acinar cells of the pancreas, hepatocytes, and bile duct epithelial cells express substantial amounts of the receptor. Osteoblast-like cells and chondrocytes also express VDR. Preimmune serum and antiserum preadsorbed with Danio VDR protein fails to detect VDR in the same tissues. The VDR is also present in the developing eye, brain, and otic vesicle of 48- and 96 h postfertilization zebrafish embryos. Parenteral administration of 1alpha,25(OH)(2)D(3) increases concentrations of VDR in intestinal epithelial cells but not in epithelial cells of the gills. Lithocholic acid, however, does not alter concentrations of VDR after parenteral administration. The data suggest that VDR is widely distributed in tissues of the zebrafish, D. rerio, and is likely to play important roles in epithelial transport, bone, and endocrine function. Furthermore, concentrations of the receptor seem to be regulated by its ligand, 1alpha,25-dihydroxyvitamin D but not by lithocholic acid. Zebrafish may serve as a useful model in which to assess the function of the VDR in diverse tissues. PMID- 18410236 TI - PDGF receptor beta is a potent regulator of mesenchymal stromal cell function. AB - Mesenchymal stromal cells (MSCs) in bone marrow are important for bone homeostasis. Although platelet-derived growth factor (PDGF) has been reported to be involved in osteogenic differentiation of MSCs, the role remains controversial and the network of PDGF signaling for MSCs has not been clarified. To clarify the underlying regulatory mechanism of MSC functions mediated by PDGF, we deleted the PDGF receptor (PDGFR)beta gene by Cre-loxP strategy and examined the role of PDGF in osteogenic differentiation of MSCs and fracture repair. In cultured MSCs, the mRNA expression of PDGF-A, -B, -C, and -D as well as PDGFRalpha and beta was detected. Depletion of PDGFRbeta in MSCs decreased the mitogenic and migratory responses and enhanced osteogenic differentiation as evaluated by increased alkaline phosphatase (ALP) activity and mRNA levels of ALP, osteocalcin (OCN), bone morphogenetic protein (BMP) 2, Runx2, and osterix in quantitative RT-PCR. PDGF-BB, but not PDGF-AA, inhibited osteogenic differentiation accompanied by decreased ALP activity and mRNA levels, except for BMP2. These effects of PDGF-BB were eliminated by depletion of PDGFRbeta in MSCs except that PDGF-BB still suppressed osterix expression in PDGFRbeta-depleted MSCs. Depletion of PDGFRbeta significantly increased the ratio of woven bone to callus after fracture. From the combined analyses of PDGF stimulation and specific PDGFRbeta gene deletion, we showed that PDGFRbeta signaling distinctively induces proliferative and migratory responses but strongly inhibits osteogenic differentiation of MSCs. The effects of PDGFRalpha on the osteogenic differentiation were very subtle. PDGFRbeta could represent an important target for guided tissue regeneration or tissue engineering of bone. PMID- 18410238 TI - Protein kinase C isozymes as potential therapeutic targets in immune disorders. AB - BACKGROUND: Members of the protein kinase C (PKC) family are key signalling mediators in immune responses, and pharmacological inhibition of PKCs may be useful for treating immune-mediated diseases. OBJECTIVE: To review and discuss the insights gained so far into various PKC isozymes and the therapeutic potential and challenges of developing PKC inhibitors for immune disorder therapy. METHODS: A literature review of the role of PKCs in immune cell signalling and recent studies describing immune functions associated with PKC isozyme deficiency in relevant mouse disease models, followed by specific case studies of current and potential therapeutic strategies targeting PKCs. RESULTS/CONCLUSION: There is vast amount of data supporting PKC isozymes as attractive drug targets for certain immune disorders. Although the development of specific PKC isozyme inhibitors has been challenging, some progress has been made. It remains to be seen if broad-scale or isozyme-selective inhibition of PKC will have clinical efficacy. PMID- 18410237 TI - Initiation of protein synthesis: a target for antimicrobials. AB - BACKGROUND: Translation initiation is a basic and universal biological process that employs significantly different components and displays substantially different mechanisms in bacterial, archaeal and eukaryotic cells. A large amount of detailed mechanistic and structural information on the bacterial translation initiation apparatus has been uncovered in recent years. OBJECTIVE: to understand which translation initiation steps could represent a novel or underexploited target for the discovery of new and specific antibacterial drugs. METHODS: Brief descriptions of the properties and mechanism of action of the major antibiotics that have a documented direct inhibitory effect on bacterial translation initiation are presented. RESULTS/CONCLUSIONS: Considerations and predictions concerning a future scenario for research and identification of bacterial translation initiation inhibitors are presented. PMID- 18410239 TI - Novel therapeutic targets in the treatment of IBD. AB - BACKGROUND: Advances in mucosal immunology have revealed a broad set of new therapeutic targets to resolve inflammation and symptoms in patients with inflammatory bowel diseases (IBD). OBJECTIVE: Despite the enormous success of anti-TNF therapies in IBD, these treatments have limited efficacy, and there continues to be concerns regarding their toxicity. Thus, a considerable unmet need exists for better treatment of these disorders. METHODS: New therapeutic targets include other pro-inflammatory cytokines such as IL-6, IL-12, IL-17 or IFN-gamma, and others. In addition, molecules directing trafficking of inflammatory cells such as integrin alpha(4)beta(7) or intracellular adhesion molecule 1 (ICAM-1) might be attractive candidates as anti-inflammatory targets. Targeting intestine-specific homing by blocking chemokine receptors such as CCR9 might provide a new avenue for treatment in the future. CONCLUSION: All these and many other different therapies are currently being investigated in IBD, the challenge will be to develop more effective therapies than those currently available. PMID- 18410240 TI - The therapeutic potential of neuronal K V 7 (KCNQ) channel modulators: an update. AB - BACKGROUND: Neuronal KCNQ channels (K(V)7.2-5) represent attractive targets for the development of therapeutics for chronic and neuropathic pain, migraine, epilepsy and other neuronal hyperexcitability disorders, although there has been only modest progress in translating this potential into useful therapeutics. OBJECTIVE: Compelling evidence of the importance of K(V)7 channels as neuronal regulatory elements, readily amenable to pharmacological modulation, has sustained widespread interest in these channels as drug targets. This review will update readers on key aspects of the characterization of these important ion channel targets, and will discuss possible current barriers to their exploitation for CNS therapeutics. METHODS: This article is based on a review of recent literature, with a focus on data pertaining to the roles of these channels in neurophysiology. In addition, I review some of the regulatory elements that influence the channels and how these may relate to channel pharmacology, and present a review of recent advances in neuronal K(V)7 channel pharmacology. CONCLUSIONS: These channels continue to be valid and approachable targets for CNS therapeutics. However, we may need to understand more about the roles of neuronal K(V)7 channels during the development of disease states, as well as to pay more attention to a detailed analysis of the molecular pharmacology of the different channel subfamily members and the modes of interaction of individual modulators, in order to successfully target these channels for therapeutic development. PMID- 18410241 TI - Upregulating substance P levels to treat obstructive sleep apnea. AB - BACKGROUND: The neuropeptide (tachykinin) substance P is widely distributed in the central and peripheral nervous systems. Substance P has been suggested to function as a neurotransmitter, cotransmitter, or neuromodulator in the central and peripheral nervous system. substance P also influences sleep physiology. Neurokinin 1 (NK-1) receptors may also be implicated in the control of sleep/wake behavior. Obstructive sleep apnea syndrome (OSAS) is defined as repeated episodes of upper airway occlusion during sleep with subsequent excessive daytime sleepiness (EDS). Substance P levels are found to be significantly lowered in patients with OSAS. OBJECTIVE: The aim of this review was to investigate the relationship between substance P, EDS and other OSAS complications. METHODS: The literature was searched using standard methods. Medline and Embase were searched systematically from 1974 to the end of February 2008 for relevant articles published in English. RESULTS/CONCLUSION: EDS seen in some OSAS patients may be associated with various pathophysiological mechanisms including changes in substance P levels. Intravenous substance P administration in OSAS patients can be effective in the treatment of EDS. Further studies on the possible relationship between low serum substance P and hypertension, reproductive function disorders, memory and learning problems in OSAS cases is required. PMID- 18410242 TI - Targeting the type 1 insulin-like growth factor receptor as a treatment for cancer. AB - BACKGROUND: The type 1 insulin-like growth factor receptor (IGF1R) plays a critical role in transformation, invasion and apoptosis protection, and is an attractive cancer treatment target. OBJECTIVE: To review IGF1R antibodies and kinase inhibitors that are in preclinical and clinical development, and to discuss questions that will influence the success of this approach in clinical practice. METHODS: This review is drawn from published literature, meeting abstracts and online resources. RESULTS/CONCLUSION: IGF1R blockade is generally well tolerated although it can induce hyperglycaemia. Single-agent activity has been documented in Ewing's sarcoma but not thus far in common solid tumours. Key issues include identification of factors that influence sensitivity to IGF1R blockade, and how most effectively to combine IGF1R inhibitors with other treatments. PMID- 18410243 TI - Targeting of extracellular proteases required for the progression of pancreatic cancer. AB - BACKGROUND: Pancreatic ductal adenocarcinoma (PDA) is the fourth leading cause of cancer-related death in the United States. Its lethality is due, in large part, to its resistance to traditional chemotherapeutics. As a result, there is an enormous effort being put into basic research to identify proteins that are required for PDA progression so that they may be specifically targeted for therapy. OBJECTIVE: To compile and analyze the evidence that suggests that extracellular proteases are significant contributors to PDA progression. METHODS: We focus on three different extracellular protease subclasses expressed in PDA: metalloproteases, serine proteases and cathepsins. Based on data from PDA and other cancers, we suggest their probable roles in PDA. RESULTS/CONCLUSIONS: Of the proteases expressed in PDA, many appear to have overlapping functions, based on the substrates they process, making therapeutics complicated. Two protease families most likely to have unique, critical functions during tumor progression, and therefore strong potential as therapeutic targets, are the a disintegrin and metalloproteases (ADAMs) and the cathepsins. PMID- 18410244 TI - Chemokines and antagonists in non-Hodgkin's lymphoma. AB - BACKGROUND: Chemokines and their receptors are a large family of molecules that control the trafficking of immune cells during their development and in response to inflammation. Non Hodgkin's lymphoma (NHL) derives from the neoplastic transformation of lymphocytes at different stages of differentiation and may show systemic, nodal and extranodal localisation as well as metastasis in different sites. OBJECTIVE: To investigate the role of the chemokine system in the pathology of NHL and as a potential drug target in this disease. METHOD: The expression of chemokines and receptors by different NHL subtypes as well as their likely functional role in terms of lymphoma tissue localisation, lymphoma growth, tumour angiogenesis and recruitment of immune cells are reviewed. The data regarding antagonists or chemokines as potential therapeutic agents for NHL is discussed. RESULTS/CONCLUSIONS: NHL's express functional chemokine receptors, which, at least in part, dictate tissue localisation and perhaps metastatic potential. Different types of chemokine antagonists have been tested in vitro and in vivo and several peptide or small molecule inhibitors have reached Phase I/II clinical trials for different diseases. Although the field is in its early days, interesting compounds are being developed, directed especially against homeostatic chemokine receptors. PMID- 18410245 TI - Small interfering RNA therapy in cancer: mechanism, potential targets, and clinical applications. AB - BACKGROUND: Small interfering RNA (siRNA) has become a powerful tool in knocking down or silencing gene expression in most cells. siRNA-based therapy has shown great promise for many diseases such as cancer. Major targets for siRNA therapy include oncogenes and genes that are involved in angiogenesis, metastasis, survival, antiapoptosis and resistance to chemotherapy. OBJECTIVES: This review briefly summarizes current advances in siRNA therapy and clinical applications in cancers, especially in pancreatic cancer. METHODS: This review article covers several aspects of siRNA therapy in cancer, which include the types of siRNA, the delivery systems for siRNA, and the major targets for siRNA therapy. Specific attention is given to siRNA in pancreatic cancer, which is our main research focus. RESULTS/CONCLUSION: siRNA can be introduced into the cells by using either chemically synthesized siRNA oligonucleotides (oligos), or vector-based siRNA (shRNA), which allows long lasting and more stable gene silencing. Nanoparticles and liposomes are commonly used carriers, delivering the siRNA with better transfection efficiency and protecting it from degradation. In combination with standard chemotherapy, siRNA therapy can also reduce the chemoresistance of certain cancers, demonstrating the potential of siRNA therapy for treating many malignant diseases. This review will provide valuable information for clinicians and researchers who want to recognize the newest endeavors within this field and identify possible lines of investigation in cancer. PMID- 18410246 TI - The P2X7 receptor as a therapeutic target. AB - BACKGROUND: The P2X7 receptor is present in a variety of cell types involved in pain, inflammatory processes and neurodegenerative conditions, thus it may be an appealing target for pharmacological intervention. The extensive use of high throughput screening (HTS) followed by a hit-to-lead (HtL) program, has prompted a number of firms to identify highly selective and metabolically stable small molecules possessing activity for both the rat and human P2X(7) receptor, which provide a novel therapeutic approach to the treatment of pain as well as neurodegenerative and inflammatory disorders. OBJECTIVE: To describe the current status of and potential for development of P2X(7) receptor-antagonists. METHODS: A literature review. RESULTS/CONCLUSIONS: We describe the recent discoveries of novel P2X(7) receptor-selective antagonists, along with their biological activity and therapeutic potential. PMID- 18410248 TI - Macromolecular modeling with rosetta. AB - Advances over the past few years have begun to enable prediction and design of macromolecular structures at near-atomic accuracy. Progress has stemmed from the development of reasonably accurate and efficiently computed all-atom potential functions as well as effective conformational sampling strategies appropriate for searching a highly rugged energy landscape, both driven by feedback from structure prediction and design tests. A unified energetic and kinematic framework in the Rosetta program allows a wide range of molecular modeling problems, from fibril structure prediction to RNA folding to the design of new protein interfaces, to be readily investigated and highlights areas for improvement. The methodology enables the creation of novel molecules with useful functions and holds promise for accelerating experimental structural inference. Emerging connections to crystallographic phasing, NMR modeling, and lower resolution approaches are described and critically assessed. PMID- 18410249 TI - Structural biology of the tumor suppressor p53. AB - The tumor suppressor protein p53 induces or represses the expression of a variety of target genes involved in cell cycle control, senescence, and apoptosis in response to oncogenic or other cellular stress signals. It exerts its function as guardian of the genome through an intricate interplay of independently folded and intrinsically disordered functional domains. In this review, we provide insights into the structural complexity of p53, the molecular mechanisms of its inactivation in cancer, and therapeutic strategies for the pharmacological rescue of p53 function in tumors. p53 emerges as a paradigm for a more general understanding of the structural organization of modular proteins and the effects of disease-causing mutations. PMID- 18410247 TI - Single-molecule studies of RNA polymerase: motoring along. AB - Single-molecule techniques have advanced our understanding of transcription by RNA polymerase (RNAP). A new arsenal of approaches, including single-molecule fluorescence, atomic-force microscopy, magnetic tweezers, and optical traps (OTs) have been employed to probe the many facets of the transcription cycle. These approaches supply fresh insights into the means by which RNAP identifies a promoter, initiates transcription, translocates and pauses along the DNA template, proofreads errors, and ultimately terminates transcription. Results from single-molecule experiments complement the knowledge gained from biochemical and genetic assays by facilitating the observation of states that are otherwise obscured by ensemble averaging, such as those resulting from heterogeneity in molecular structure, elongation rate, or pause propensity. Most studies to date have been performed with bacterial RNAP, but work is also being carried out with eukaryotic polymerase (Pol II) and single-subunit polymerases from bacteriophages. We discuss recent progress achieved by single-molecule studies, highlighting some of the unresolved questions and ongoing debates. PMID- 18410250 TI - Who develops severe malaria? Impact of access to healthcare, socio-economic and environmental factors on children in Yemen: a case-control study. AB - OBJECTIVE: To investigate the impact of socio-economic and environmental factors on developing severe malaria in comparison with mild malaria in Yemen. METHOD: Case-control study comparing 343 children aged 6 months to 10 years diagnosed with WHO-defined severe malaria (cases) at the main children's hospital in Taiz and 445 children with mild malaria (controls) diagnosed in the health centres, which serve the areas where the cases came from. RESULTS: In univariate analysis, age <1 year, distance from health centre, delay to treatment and driving time to health centre were associated with progression from mild to severe malaria. In multivariate analysis, distance to nearest health centre >2 km was significantly associated with progression to severe disease. Environmental and vector control factors associated with protection from acquiring malaria (such as sleeping under bednets) were not associated with protection from moving from mild to severe disease. CONCLUSIONS: Innovative ways to improve access to antimalarial treatment for those living more then 2 km away from health centres such as home management of malaria, especially for infants and young children, should be explored in malaria-endemic areas of Yemen. PMID- 18410252 TI - Autologous serum eyedrops: literature review and implications for transfusion medicine specialists. AB - Persistent corneal epithelial defects (CEDs) are caused by many diseases that are usually associated with decreased production of tears or reduced corneal sensitivity. Although surgical treatments are available for severe cases, CEDs are still difficult for ophthalmologists to treat. One treatment for CEDs that is uncommonly used in the United States is autologous serum eyedrops (ASEs). The first application of ASEs was described in 1984. This landmark report was followed by multiple studies that carefully evaluated the epithelial-promoting properties of ASEs and refined the means of ASE preparation. A number of clinical studies suggested the efficacy of ASEs in various ophthalmologic conditions. This article reviews the efficacy and complications of ASE use reported in these studies. Given that ophthalmologists may consult the blood bank to request ASEs, transfusion medicine physicians should be aware of the issues related to ASE preparation, storage, and potential utility. PMID- 18410251 TI - Auto-antibodies to receptor tyrosine kinases TrkA, TrkB and TrkC in patients with chronic Chagas' disease. AB - The Chagas' disease parasite Trypanosoma cruzi promotes survival and differentiation of neurones by binding and activating nerve growth factor (NGF) receptor TrkA. The functional mimic of NGF in T. cruzi is a surface-bound and shed immunogenic protein [neurotrophic factor/trans-sialidase (TS)], which raised the possibility that immune response to T. cruzi in general and to neurotrophic factor/TS in particular leads to loss of immunological tolerance to host NGF and/or the NGF-binding partner TrkA. In testing this hypothesis, we found that sera of individuals with chronic Chagas' disease bear unique IgG2 autoantibodies that bind TrkA and TrkA family members TrkB and TrkC (ATA). Binding of ATA to Trk receptors is specific because the autoantibodies did not cross-react with five other growth factor receptors, NGF and other neurotrophins, and T. cruzi. Thus, individuals with chronic Chagas' disease produce unique antibodies that react with pan-Trk receptors, one of which (TrkA) T. cruzi exploits to inhibit host cell apoptosis and to promote cellular invasion. PMID- 18410253 TI - Plasmapheresis in asparaginase-induced hypertriglyceridemia. AB - BACKGROUND: Asparaginase is an essential component of the chemotherapy regimens during the induction and intensification phases for acute lymphoblastic leukemia and lymphoblastic lymphoma. The administration of asparaginase can cause elevation of the triglyceride levels. CASE REPORT: This study reports the case of an adult patient presenting with acute pancreatitis secondary to severe hypertriglyceridemia after asparaginase treatment during the consolidation phase of lymphoblastic lymphoma. This condition was successfully treated with plasmapheresis. CONCLUSIONS: Our case demonstrates that plasmapheresis in severe cases of hypertriglyceridemia offers a safe and efficient treatment option for symptomatic patients. PMID- 18410254 TI - Inactivation of parvovirus B19 during STIM-4 vapor heat treatment of three coagulation factor concentrates. AB - BACKGROUND: To enhance the viral safety margins, nanofiltration has been widely integrated into the manufacturing process of plasma-derived medicinal products. Removal of smaller agents such as parvovirus B19 (B19V) by filtration, however, is typically less efficient. Because recent investigations have demonstrated that B19V may be more heat sensitive than animal parvoviruses, the potential B19V inactivation by a proprietary vapor heating procedure (STIM-4) as incorporated into the manufacturing processes of several nanofiltered coagulation factor concentrates was investigated. STUDY DESIGN AND METHODS: An infectivity assay based on quantitative reverse transcription-polymerase chain reaction (TaqMan, Applied Biosystems) detection of B19V mRNA after inoculation of a permissive cell line (UT7 Epo S1 cells) was used to investigate the virus inactivation capacity of the STIM-4 vapor heat treatment as used during the manufacture of nanofiltered second-generation Factor VIII inhibitor-bypassing activity (FEIBA), F IX complex, and FVII products. RESULTS: In contrast to animal parvoviruses, both B19V genotypes investigated, that is, 1 and 2, were shown to be surprisingly effectively inactivated by the STIM-4 vapor heat treatment process, with mean log reduction factors of 3.5 to 4.8, irrespective of the product intermediate tested. CONCLUSION: The newly demonstrated effective inactivation of B19V by vapor heating, in contrast to the earlier used animal parvoviruses, results in significant B19V safety margins for STIM-4-treated coagulation factor concentrates. PMID- 18410255 TI - Observational studies on the effect of dietary antioxidants on asthma: a meta analysis. AB - BACKGROUND AND OBJECTIVE: It has been suggested that the rapid increase in asthma prevalence may in part be due to a decrease in the intake of dietary antioxidants, including vitamin C, vitamin E and beta-carotene. Epidemiological studies investigating the association between dietary antioxidant intake and asthma have generated inconsistent results. A meta-analysis was undertaken to examine the association between dietary antioxidant intake and the risk of asthma. METHODS: The MEDLINE database was searched for observational studies in English-language journals from 1966 to March 2007. Data were extracted using standardized forms. Pooled odds ratios (OR) with 95% confidence intervals (CI) were calculated using a random effects model. Ten studies were eligible for inclusion. Seven studies, comprising 13 653 subjects, used asthma or wheeze as their outcome; three studies explored the effect of antioxidant intake on lung function. RESULTS: A higher dietary intake of antioxidants was not associated with a lower risk of having asthma. The pooled OR for having asthma were 1.06 (95% CI: 0.79-1.43) for subjects with a higher dietary vitamin C intake compared with those with a lower intake; 0.88 (95% CI: 0.61-1.25) for vitamin E; and 1.12 (95% CI: 0.77-1.62) for beta-carotene. There was no significant association between dietary antioxidant intake and lung function except for a positive association between vitamin C intake and an increase in FEV(1) (29.1 mL, 95% CI: 0.4-58.6, P = 0.05). CONCLUSIONS: This meta-analysis does not support the hypothesis that dietary intake of the antioxidants vitamins C and E and beta carotene influences the risk of asthma. PMID- 18410256 TI - Diagnosis of pulmonary aspiration: a mouse model using a starch granule test in bronchoalveolar lavage. AB - BACKGROUND AND OBJECTIVE: Pulmonary aspiration (PA) is a significant respiratory disease in children. However, the diagnosis of aspiration is often difficult owing to the poor efficacy of currently available diagnostic tests. The aim of this study was to assess in a mouse model the specificity of starch granule detection in BAL as a new method for detecting PA in children. METHODS: Twenty BALB/c mice were divided into the following groups according to the solution instilled into the airways: corn flour milk 7.5%-a source of starch (CM), Pseudomonas aeruginosa, normal saline and a control group. BAL was performed 2 days after instillation. Detection of starch granules and lipid-laden macrophages were compared in BAL. RESULTS: Starch granules were detected in BAL fluids from all mice in the CM group (food aspiration model), whereas no starch granules were detected in the other three groups, demonstrating a sensitivity and specificity of 100%. On the other hand, lipid-laden macrophages were found in all mice from all the groups studied. CONCLUSIONS: The detection of starch granules in BAL is a simple and highly specific method for the diagnosis of PA in an experimental model. Clinical studies using the starch granule detection method in BAL should be tested in at risk patients to evaluate the utility of this method for investigating PA. PMID- 18410257 TI - Predictors for wheezing phenotypes in the first decade of life. AB - BACKGROUND AND OBJECTIVE: This study examined prenatal, perinatal and early childhood predictors of wheezing phenotypes in the first decade of life. METHODS: Information on current wheezing, was collected prospectively from five surveys conducted every 2 years over the first decade of life. Five wheezing phenotypes were defined: non-wheezers, preschool, primary-school, intermittent and persistent wheezers. Logistic regression with adjustment for survey design was used to determine the predictors of wheezing phenotypes. RESULTS: Data on 2711 children were used in the analysis. Early respiratory infection, the child's allergy and parental asthma were significant risk factors for preschool, intermittent and persistent wheeze. The child's allergy and parental asthma had stronger associations with persistent wheeze than with preschool wheeze. Breastfeeding was a significant predictor of both preschool and intermittent wheezing. Daycare attendance was a risk factor for preschool wheeze but a protective factor for primary-school wheezing. Crowding at home was a protective factor for both preschool and primary-school wheeze. Parental smoking was a significant factor for preschool wheeze. CONCLUSION: This study identified different predictors for each wheezing phenotype with some degree of overlap. The observed differential effects for these conditions raises the possibility that there are different aetiologies for asthma among children. PMID- 18410258 TI - Neutrophil elastase activity in acute lung injury and respiratory distress syndrome. AB - BACKGROUND AND OBJECTIVE: Neutrophil elastase (NE) may play a key role in the development of acute lung injury (ALI) or ARDS. NE activity (NEA) was measured in patients with ALI treated with a selective NE inhibitor. METHODS: NEA and NE alpha1-antitrypsin (NE-AT) complex were measured in plasma before, during and after the administration of the selective NE inhibitor, sivelestat, in 32 patients with a diagnosis of ALI or ARDS. NEA index (NEAI) was calculated as NEA/NE-AT. The sequential organ failure assessment (SOFA) score and the ratio PaO(2)/fraction of inspired oxygen (FiO(2)) were measured. RESULTS: NEA and NE-AT was raised in all patients. Sivelestat reduced NEAI and NEA (P < 0.01 for both) but not NE-AT and NEA, and NEAI returned to pretreatment levels. NEA correlated closely with NE-AT before, but not after treatment. No relationship was observed between these indices and SOFA score or PaO(2)/FiO(2) ratio. CONCLUSIONS: Sivelestat reduced NEA and NEAI in patients with ALI or ARDS suggesting NE inhibition. A larger study is needed to determine the relationship of NEA, NE-AT and NEAI with the outcome of ALI/ARDS. PMID- 18410259 TI - Lung cancer in patients with chronic pyothorax. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to describe the features of lung cancers associated with chronic tuberculous pyothorax. METHODS: Clinicopathological data from patients with coexisting lung cancer and chronic latent pyothorax caused by tuberculosis (TB) were analysed, and cancer tissue samples were investigated for the presence of Epstein-Barr virus. RESULTS: Twelve patients were identified, and all had a history of tuberculous pleuritis or surgical intervention for TB. The interval between the onset of TB and lung cancer was more than 30 years in nine patients and the most frequent symptom was chest pain (six patients). All cancers were in the ipsilateral lung to the pyothorax, and in nine of the 12 patients the cancers were located adjacent to the pyothorax. In situ hybridization analysis for Epstein-Barr virus-encoded small RNA failed to show positive signals in any of the six cancer tissues examined. CONCLUSIONS: Lung cancer associated with chronic pyothorax always developed in the ipsilateral lung to the pyothorax, and there was no evidence for the presence of Epstein-Barr virus in the cancer tissues examined. PMID- 18410260 TI - Common functional polymorphisms in the cathepsin S promoter in Japanese subjects: possible contribution to pulmonary emphysema. AB - BACKGROUND AND OBJECTIVE: Cathepsin S is involved in the pathogenesis of COPD in murine models overexpressing interferon (IFN)-gamma and IL-13. It is widely accepted that genetic factors partly influence susceptibility to COPD; however, the association of genetic polymorphisms in the cathepsin S gene with COPD has not been reported previously. In this study, functional polymorphisms in the 5' flanking region of the human cathepsin S gene were identified and their association with COPD phenotypes was investigated. METHODS: Genetic polymorphisms were detected by single strand conformation polymorphism analysis and direct sequencing. Genotypes and haplotypes were determined using amplification refractory mutation systems and single strand conformation polymorphism analysis. A promoter-reporter assay was used to compare transcriptional activities between the haplotypes on stimulation with IFN-gamma. Haplotype analysis was performed on 204 smoking and 104 non-smoking Japanese subjects. RESULTS: Four novel polymorphisms were detected in the 5'-flanking region and three major haplotypes were identified. The haplotypes were associated with distinct promoter activities on stimulation with IFN-gamma and there was a trend indicating that higher transcriptional activity was linked to less pulmonary emphysema. CONCLUSION: Novel functional polymorphisms were identified in the cathepsin S gene, which has a possible association with pulmonary emphysema in the Japanese population. PMID- 18410261 TI - Evaluation of the anti-inflammatory effect of infliximab in a mouse model of acute asthma. AB - BACKGROUND AND OBJECTIVE: To evaluate the potential role of anti-tumour necrosis factor (TNF)-alpha mAb (infliximab) on the inflammatory response in a mouse model of acute asthma. METHODS: BALB/c mice received intraperitoneal (i.p.) ovalbumin (OVA) on days 0 and 14, 100 microg of OVA intranasally on day 14 and 50 microg of OVA intranasally on days 25, 26 and 27. The low-dose (2.5 mg/kg) and high-dose (6.25 mg/kg) infliximab groups received i.p. infliximab before each i.p. sensitization and on challenge days 1, 6, 13, 20 and 27. The control group received i.p. injections of normal saline with alum on days 0 and 14 and normal saline without alum on days 14, 25, 26 and 27. RESULTS: There were statistically significant decreases in the numbers of BAL fluid (BALF) neutrophils, eosinophils, as well as lung eosinophils in both the low- and high-dose infliximab groups when compared with the control OVA sensitized/challenged group. The lower dose of infliximab did not alter lung neutrophil counts, but a marked decrease was seen with the high dose of infliximab. After treatment with low and high doses of infliximab, BALF levels of regulated on activation normal T cell expressed and secreted (RANTES), granulocyte macrophage-colony stimulating factor (GM-CSF), TNF-alpha, IL-6, macrophage inflammatory protein (MIP)-2, and levels of RANTES, IL-4, GM-CSF, TNF-alpha, IL-6 and MIP-2 in lung tissue were significantly decreased when compared with the control OVA sensitized/challenged group. There was a significant decrease in BALF IL-4 only in the high-dose infliximab group. CONCLUSIONS: These results show that an anti-TNF-alpha mAb has a considerable anti-inflammatory effect on allergen-induced lung inflammation in an animal model of acute asthma. PMID- 18410262 TI - Airway inflammation in employees involved in cultivating Japanese mushrooms (bunashimeji). AB - BACKGROUND AND OBJECTIVE: Chronic inhalation of spores may cause respiratory symptoms such as productive cough and sputum. The purpose of this study was to determine the clinical pathophysiology of airway inflammation caused by bunashimeji spores and to investigate whether the spores have direct toxic inflammatory effects. METHODS: Sensitized employees with respiratory symptoms and a stimulation index (SI) > 200%, and non-sensitized employees with a SI < 200% were enrolled. They underwent sputum induction and chest high-resolution computed tomography (HRCT). The in vitro effect of bunashimeji spore solutions on normal human bronchial epithelial (NHBE) cell cultures was investigated using the air liquid interface method. Bunashimeji spore solution was added at 10(4) or 10(6) spores per 20 microL/well. The interleukin (IL)-8 and epithelial neutrophil activating peptide-78 (ENA-78) concentrations in the medium and IL-8 mRNA expression of NHBE cells were assessed after each stimulation. RESULTS: Sensitized employees were divided into 14 with normal HRCT and 9 with abnormal HRCT. Fifteen of the sensitized group and five of the non-sensitized group had a productive cough and sputum. The neutrophil counts in induced sputum were significantly higher in subjects with abnormal HRCT than in those with normal HRCT. IL-8 and ENA-78 concentrations following stimulation with 10(4) and 10(6) spores were significantly increased compared with PBS only on day 9. IL-8 mRNA expression due to spore stimulation was significantly increased compared with control. IL-8 mRNA expression with 10(6) spore stimulation was significantly increased on days 6 and 12 compared with 10(4) spores. CONCLUSION: The inhalation of spores directly produces toxic inflammatory effects in the airways, independent of the degree of sensitization. PMID- 18410263 TI - Poor interpretation of pulmonary function tests in patients with concomitant decreases in FEV1 and FVC. AB - BACKGROUND AND OBJECTIVE: A new interpretative strategy for pulmonary function tests (PFT) has been proposed by the American Thoracic Society/European Respiratory Society (ATS/ERS) Task Force. To assess the accuracy of this strategy, clinical diagnosis was compared with the PFT interpretation in patients showing concomitant decreases in FEV(1) and FVC. METHODS: A retrospective study was conducted of 681 patients with normal FEV(1)/FVC and low FVC who underwent lung volume measurements and spirometry on the same date between July and November 2005 at Asan Medical Center, Seoul, Korea. Patients were clinically diagnosed by the consensus decision of two respiratory physicians, and the kappa coefficient was calculated to compare the clinical diagnosis with the PFT interpretation using the ATS/ERS strategy. RESULTS: The PFT interpretation showed an obstructive pattern in 205 patients and a restrictive pattern in 476. Of the 205 patients with an obstructive pattern on PFT, 44 were clinically diagnosed with obstructive, 97 with restrictive and 17 with mixed disease, whereas 47 patients had no disease. Of the 476 patients with a restrictive pattern on PFT, 11 were clinically diagnosed with obstructive, 369 with restrictive and 60 with mixed disease, whereas 36 patients had no disease. The kappa coefficient was 0.35 (95% confidence interval: 0.26-0.44; P < 0.0001). CONCLUSIONS: The weak agreement between the clinical diagnosis and the PFT interpretation in patients showing concomitant decreases in FEV(1) and FVC suggests that other clinical findings should be assessed in addition to PFT. PMID- 18410264 TI - More than a gut feeling: the human visceral brain re-visited. PMID- 18410265 TI - Oesophageal afferent pathway sensitivity in non-erosive reflux disease. AB - Patients with non-erosive reflux disease (NERD) report symptoms which commonly fail to improve on conventional antireflux therapies. Oesophageal visceral hyperalgaesia may contribute to symptom generation in NERD and we explore this hypothesis using oesophageal evoked potentials. Fifteen endoscopically confirmed NERD patients (four female, 29-56 years) plus 15 matched healthy volunteers (four female, 23-56 years) were studied. All patients had oesophageal manometry/24-h pH monitoring and all subjects underwent evoked potential and sensory testing, using electrical stimulation of the distal oesophagus. Cumulatively, NERD patients had higher sensory thresholds and increased evoked potential latencies when compared to controls (P = 0.01). In NERD patients, there was a correlation between pain threshold and acid exposure as determined by DeMeester score (r = 0.63, P = 0.02), with increased oesophageal sensitivity being associated with lower DeMeester score. Reflux negative patients had lower pain thresholds when compared to both reflux positive patients and controls. Evoked potentials were normal in reflux negative patients but significantly delayed in the reflux positive group (P = 0.01). We demonstrate that NERD patients form a continuum of oesophageal afferent sensitivity with a correlation between the degree of acid exposure and oesophageal pain thresholds. We provide objective evidence that increased oesophageal pain sensitivity in reflux negative NERD is associated with heightened afferent sensitivity as normal latency evoked potential responses could be elicited with reduced afferent input. Increased oesophageal afferent pain sensitivity may play an important role in a subset of NERD and could offer an alternate therapeutic target. PMID- 18410266 TI - Central and peripheral role of the nociceptin/orphaninFQ system on normal and disturbed colonic motor function and faecal pellet output in the rat. AB - In this study, seeking further information on the role of the nociceptin/orphanin FQ (N/OFQ)-ergic system in normal and disturbed colonic motor function in rats, we compared the colonic effects of UFP-112, a novel highly potent agonist, with those of N/OFQ. When injected intracerebroventricularly (i.c.v.) and intraperitoneally (i.p.), UFP-112 and N/OFQ increased bead expulsion time in a statistically significant and dose-related manner and reduced the percentage of rats with castor oil-induced diarrhoea. UFP-112 showed greater efficacy, higher potency and longer-lasting inhibitory effects than N/OFQ, and pretreatment with UFP-101, a selective antagonist, blocked the N/OFQ analogue-induced responses in both tests. When injected i.c.v., UFP-112 and N/OFQ inhibited corticotrophin releasing factor- and restrain stress-stimulated faecal pellet excretion significantly and in a dose-related manner. Conversely, when injected peripherally both peptides significantly inhibited colonic propulsive motility but did so in a non-dose-related manner. In conclusion, these findings indicate that, in the rat, the central and peripheral N/OFQ systems have an inhibitory role in modulating distal colonic propulsive motility under physiological and pathological conditions. UFP-112 therefore promises to be a useful pharmacological tool for investigating the role of the N/OFQ system in motor functions in the distal colonic tract under physiological and pathological conditions. PMID- 18410267 TI - Clinicopathological and immunohistochemical findings in an autopsy case of tuberous sclerosis complex. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant, multisystem disorder caused by mutations in either the TSC1 or TSC2 genes and characterized by developmental brain abnormalities. In the present study we discuss the neuropathological findings of a 32-year-old patient with a germ-line mutation in the TSC2 gene. Post mortem MRI combined with histology and immunocytochemical analysis was applied to demonstrate widespread anatomical abnormalities of gray and white matter structure. TSC brain lesions were analyzed for loss of heterozygosity (LOH) on chromosome 16p13. The neuropathological supratentorial abnormalities were represented by multiple subependymal nodules (SENs) and cortical tubers. In addition to cerebral cortical lesions, cerebellar lesions and hippocampal sclerosis were also observed. LOH was not found in the cortical tubers and SENs of this patient. Immunocytochemical analysis of the TSC brain lesions confirmed the cell-specific activation of the mTOR pathway in cortical tubers, SENs and cerebellum, as well as differential cellular localization of hamartin and tuberin, the TSC1 and TSC2 gene products. Examination of the pathological brain regions revealed activated microglial cells and disruption of blood-brain barrier permeability. Predominant intralesional cell-specific distribution was also detected for the multidrug transporter protein P-gp, possibly explaining the mechanisms underlying the pharmacoresistance to antiepileptic drugs. Autopsy findings confirm the complexity of the brain abnormalities encountered in TSC patients and proved useful in clarifying certain aspects of the pathogenesis, epileptogenesis and pharmacoresistance of TSC lesions. PMID- 18410268 TI - Peripheral and multiple cranial neuropathies in a 74-year-old woman. PMID- 18410269 TI - An autopsy case of spinal muscular atrophy type III (Kugelberg-Welander disease). AB - We report an autopsy case of a 67-year-old man clinicogenetically diagnosed as having spinal muscular atrophy (SMA) type III (Kugelberg-Welander disease), showing slowly progressive muscle wasting and weakness of the extremities. His brother showed similar manifestations. Autopsy revealed neuronal loss and severe gliosis in the anterior horns of the spinal cord, a marked neurogenic change of skeletal muscles and mild degeneration of cardiomyocytes. Chromatolytic change was seen in the anterior horn, but not in the Clarke's and thalamic nuclei. The anterior spinal roots were atrophic, and there was loss of myelinated fibers with abundant glial bundles. In addition, degeneration was also observed in the posterior column and dentate nucleus. The pathological features were essentially similar to those of SMA I. Chronic change was prominent while acute change was mild in degree, corresponding to a very long clinical course. PMID- 18410270 TI - Malignant non-teratoid medulloepithelioma of ciliary body with retinoblastic differentiation: a case report and review of literature. AB - We present a 6-year-old child with intraocular and extraocular mass and high intraocular pressure. The tumor mass involved a disorganized anterior segment and extended through the medial cornea and sclera. A preliminary diagnosis of retinoblastoma with extraocular extension was made. An exenteration of the left globe and orbital tissue was performed. Histological examination showed that the lesion, which occupied the posterior chamber, involved the ciliary body, extending into the iris, sclera and cornea, projecting beyond the cornea anteriorly and extending to the retina posteriorly. The tumor cells were diffusely immunoreactive to vimentin, neuron specific enolase and CD 138. The medulloepithelioma cells were focally positive to cytokeratin (AE1/AE3), cytokeratin 18, CD56,CD57, S100, HMB-45 and bcl2 while areas of retinoblastic differentiation showed diffuse immunoreactivity to synaptophysin, neurofilament and CD138 with focal immunoreactivity to calretinin. All tumor cells showed no immunoreactivity to cytokeratin 7, cytokeratin 20, epithelial membrane antigen, carcinoembryonic antigen, desmin, GFAP, and chromogranin. Nuclear staining for P53 was seen in 80% of tumor cells. The ki-67 index was 90%. The tumor was described as malignant intraocular non-teratoid medulloepithelioma with retinoblastic differentiation arising from the ciliary body. Tumor satellites were seen in the adjacent periocular soft tissue. The treatment involved exenteration of the left globe and orbital tissue with secondary skin graft following chemotherapy. The patient is well and has no recurrence after 1 year of treatment. We report that medulloepithelioma can present as a case of infantile glaucoma, can show signs of intraocular calcifications and can show retinoblastic differentiation. PMID- 18410271 TI - Paraganglioma in pituitary fossa. AB - Paragangliomas are neuroendocrine tumors of paraganglionic tissue which are extremely rare in the sellar area. We present a case of sellar paraganglioma with parasellar and suprasellar extension in a 70-year-old man who presented with headache. CT demonstrated sellar mass with suprasellar and right parasellar extension. The lesion was removed subtotally via a trans-sphenoidal approach. Histopathological diagnosis was paraganglioma. Differential diagnosis between paragangliomas and unusual types of pituitary adenomas, especially null-cell adenomas, is made by presence of cell nests (Zellballen) in paraganglioma, lack of immunopositivity for pituitary hormones, cytoplasmic immunopositivity for chromogranin A and neuron-specific enolase, and immunonegativity for cytokeratin 19, cytokeratin 20 and endomysial antibody. In the case of a mass in the sellar region, paraganglioma, although rare, should be included in the differential diagnosis. PMID- 18410272 TI - Marinesco-Sjogren syndrome with atrophy of the brain stem tegmentum and dysplastic cytoarchitecture in the cerebral cortex. AB - Marinesco-Sjogren syndrome (MSS) is a progressive multisystem disease with autosomal recessive inheritance characterized by cataracts, mental retardation, and cerebellar ataxia. Recently, two causative genes for MSS, SIL1 and SARA2, have been identified. On the other hand, the histopathologic features of the CNS in this syndrome have not yet been clarified in detail. We report here the features of an autopsy case of MSS with progressive myopathy, in which atrophy of the cerebellum and brain stem tegmentum, retinal degeneration, and dysplastic cytoarchitecture in the cerebral cortex were evident. An elder brother of the patient showed quite similar symptoms, implying an autosomal recessive mode of inheritance. However, we detected no mutations in the available genes. This case appears to represent an unusual example of MSS manifesting widespread developmental anomaly and neuronal degeneration in the CNS. PMID- 18410273 TI - An autopsy case of Fabry disease with neuropathological investigation of the pathogenesis of associated dementia. AB - The pathogenesis of dementia associated with Fabry disease was examined neuropathologically in an autopsy case. The patient was a 47-year-old computer programmer who developed renal failure at the age of 36, necessitating peritoneal dialysis, and thereafter suffered in succession episodic pulmonary congestion, bradyacusia, heart failure, and dementia, before dying of acute myocardial infarction. MRI of the brain demonstrated leuko-araiosis. The CNS parenchyma showed widespread segmental hydropic swelling of axons in the bilateral cerebral and cerebellar deep white matter in addition to neuronal ballooning due to glycolipid storage in a few restricted nuclei and multiple tiny lacunae. Hydropic axonal swelling was also sparsely distributed in the pyramidal tract, pedunculus cerebellaris superior and brachium colliculi inferioris, but wallerian degeneration of these tracts was absent. Additional features included angiopathy of the subarachnoidal arteries due to Fabry disease, such as medial thickening resulting from glycolipid deposition in smooth muscle cells (SMCs) and adventitial fibrosis with lymphocytic infiltration, together with widespread subtotal or total replacement of medial SMCs by fibrosis, associated with prominent intimal fibrous thickening and undulation of the internal elastic membrane of medium-sized (1000-100 microm diameter) arteries. The findings in this case suggest that axonopathic leukoencephalopathy due to multisegmental hydropic swelling of axons in the bilateral cerebral deep white matter is responsible for the dementia associated with Fabry disease, and may be caused by ischemia resulting from widespread narrowing and stiffening of medium-sized subarachnoidal arteries and progressive heart failure. PMID- 18410274 TI - Presence of crystalline inclusions in the peripheral nerve of a patient with IgA lambda monoclonal gammopathy of undetermined significance. AB - Association of a peripheral neuropathy with an IgA monoclonal gammopathy of undetermined significance (MGUS) is not commonly observed and is sometimes considered as coincidental. We present a case in which the nerve biopsy revealed the presence of crystalline inclusions in the endoneurium, a very unusual finding. A 75-year-old man complained of paresthesiae in both feet and unsteady gait for 6 months. He had no weakness, but deep tendon reflexes were absent and vibratory sensation distally diminished in both legs. An IgA lambda MGUS was evidenced in his serum at 10.2 g/L with 7% plasma cells in his bone marrow and no lytic lesion at skeletal examination. A superficial peroneal nerve biopsy was performed and showed numerous crystalline inclusions in the endoneurium. These were located in the cytoplasm of macrophagic histiocytes or free in the vicinity of nerve fibers. There was also a marked loss of myelinated nerve fibers and several "onion bulb" formations surrounding either isolated remyelinating fibers or small clusters of remyelinating fibers. Such crystalline inclusions have mainly been observed in the cytoplasm of plasma cells in cases of multiple myeloma, and correspond to non-secreted IgA or IgG immunoglobulins with a kappa or rarely lambda light chain. Such inclusions have also been reported in the cytoplasm of the epithelial cells from corneal fragments, in patients with multiple myeloma or IgG MGUS, and in the tubular cells from the kidney of patients with multiple myeloma and a nephrotic syndrome. In the literature, there is only one very briefly mentioned case of neuropathy associated with a myeloma and with crystalline inclusions present in the epineurium. Thus, in dysglobulinemic neuropathy, nerve fibers can be damaged by three kinds of interstitial deposits, easily identified by immunohistochemistry and at ultrastructural examination: the well known amyloid fibrils, granulo-fibrillar deposits and also crystalline inclusions. PMID- 18410275 TI - A case of metaplastic meningioma with extensive xanthomatous change. AB - We report a case of metaplastic meningioma with extensive xanthomatous change occurring in a 61-year-old woman. Magnetic resonance imaging demonstrated a well demarcated, dura-based mass measuring 7 cm in maximal diameter in the right occipital to parietal area. Under a clinical diagnosis of meningioma, right parietal craniotomy was performed. Histologically, the tumor showed extensive xanthomatous change together with the common features of meningothelial meningioma. Tumor cells showed diffuse xanthomatous change in various quantities, and xanthomatous tumor cells frequently showed a gradual transition to the non xanthomatous counterpart. The xanthomatous tumor cells showed immunopositivity for epithelial membrane antigen (EMA), vimentin, fatty acid synthase and several histiocytic markers (CD68, Ki-M1p, MAC387, lysozyme, alpha 1-antitrypsin and alpha 1-antichymotrypsin). In addition, there was patchy infiltration of foamy macrophages. Since these histiocytes did not show immunopositivity for EMA, these cells were distinguished from the xanthomatous tumor cells. These two types of xanthomatous changes comprised approximately two-thirds of the whole tumor tissue. Any atypical features such as necrosis, frequent mitotic figures or brain parenchymal invasion were not observed. We diagnosed this case as xanthomatous meningioma, WHO grade I. PMID- 18410276 TI - Neuromyelitis optica/Devic's disease: gene expression profiling of brain lesions. AB - Neuromyelitis optica (NMO), also known as Devic's disease, is an inflammatory demyelinating disease that affects selectively the optic nerves and the spinal cord, possibly mediated by an immune mechanism distinct from that of multiple sclerosis (MS). Recent studies indicate that NMO also involves the brain. Here, we studied gene expression profile of brain lesions of a patient with NMO by using DNA microarray, along with gene expression profile of the brains of Parkinson disease and amyotrophic lateral sclerosis patients. We identified more than 200 genes up-regulated in NMO brain lesions. The top 20 genes were composed of the molecules closely associated with immune regulation, among which marked up regulation of interferon gamma-inducible protein 30 (IFI30), CD163, and secreted phosphoprotein 1 (SPP1, osteopontin) was validated by real time RT-PCR, Northern blot and Western blot analysis. Pathologically, CD68(+) macrophages and microglia expressed intense immunoreactivities for IFI30 and CD163 in NMO lesions, consisting of inflammatory demyelination, axonal loss, necrosis, cavity formation, and vascular fibrosis. KeyMolnet, a bioinformatics tool for analyzing molecular interaction on the curated knowledge database, suggested that the molecular network of up-regulated genes in NMO brain lesions involves transcriptional regulation by the nuclear factor-kappaB (NF-kappaB) and B lymphocyte-induced maturation protein-1 (Blimp-1). These results suggest that profound activation of the macrophage-mediated proinflammatory immune mechanism plays a pivotal role in development of NMO brain lesions. PMID- 18410277 TI - The activation of ERK1/2 MAP kinases in glioblastoma pathobiology and its relationship with EGFR amplification. AB - The ERK1/2 activated protein kinase (MAPK) pathway is a critical signaling system that mediates ligand-stimulated signals for the induction of cell proliferation, differentiation and survival, involved in malignant transformation. The purpose of this study was to determine the activation of ERK1/2 in this tumor, and to determine the relationship of ERK1/2 activation with the amplification/overexpression of EGFR as well as with 9p21 locus gene alterations, both of which are genetic factors frequently associated with glioblastoma. We used immunohistochemistry and Western blot analysis to analyze the activation of ERK1/2 in 22 patients with glioblastoma, and we studied the amplification/overexpression of EGFR; as well as the molecular alterations in 9p21 locus genes. Positive immunostaining ERK1/2 was observed in 86.4% of the tumors, displaying mainly nuclear immunolocalization; and by immunoblotting, ERK1/2 was activated in 68% of the cases. The 70% of cases with EGFR amplification presented activated ERK1/2. The joint presence of amplified EGFR and alterations in the 9p21 genes was observed in 50% of the cases, whereas the simultaneous occurrence of these two phenomena with the activation of ERK1/2 was observed in 40% of the cases. Our results suggest that the activation of ERK1/2 is implicated in the pathobiology of glioblastoma. This activation of ERK1/2 is probably related in part to the amplification of EGFR as well as to alterations in 9p21 locus genes (homozygous deletion and promoter methylation). However, the activation of ERK1/2 also involves pathways that are independent of the EGFR. PMID- 18410278 TI - Bcl-2 expression in a papillary tumor of the pineal region. AB - Neuroepithelial papillary tumor of the pineal region (PTPR) has been described by several groups and recognized by the 2007 World Health Organization Classification of Tumors of the Central Nervous System. The proto-oncogen Blc-2 can function as an apoptosis suppressor and can promote neoplastic transformation. It may also be involved in neuroendocrine differentiation in some tumors. As PTPRs express neuroendocrine markers, we investigated the expression of Bcl-2 in tumoral cells of a new case of PTPR in a 42-year-old woman. Bcl-2 immunostaining was detected in the cytoplasm of the tumoral cells; staining intensity was heterogeneous from cell to cell and more intense in papillary areas. This intense expression of Bcl-2 in one case of PTPR with a high proliferation index (8%) might be related to the malignancy of this neoplasm. It will be interesting to investigate the prognosis impact of Bcl-2 expression in a large series of PTPRs. PMID- 18410279 TI - An atypical autopsy case of Lewy body disease with clinically diagnosed major depression: a clinical, radiological and pathological study. AB - We report an 84-year-old woman who was clinically diagnosed with late-life major depression (LLMD) and having a diffuse type of dementia with Lewy bodies (DLB) neuropathologically. Clinically, this case showed depressive mood, anxiety, and irritation, but did not show cognitive dysfunction, visual hallucination, fluctuation of alertness and parkinsonism, which define the criteria for diagnosing DLB. Neuropathological examination demonstrated abundant Lewy-related pathology including Lewy bodies and neurites in the hippocampal region and the cerebral cortex, and moderate levels in brain stem nuclei including the substantia nigra, locus ceruleus and dorsal raphe nucleus. These findings suggest the possibility that Lewy-related pathology is associated with the depressive symptoms. Furthermore, it must be noted that some patients diagnosed with LLMD clinically may develop pathology of DLB without the typical or usual clinical symptoms. PMID- 18410280 TI - MM2-cortical-type sporadic Creutzfeldt-Jakob disease with early stage cerebral cortical pathology presenting with a rapidly progressive clinical course. AB - We report the case of a 67-year-old man with MM2-cortical-type sporadic Creutzfeldt-Jakob disease (sCJD) with a rapidly progressive clinical course of 5 months. Initial symptoms were progressive memory disturbance and dementia. MRI revealed high signal-intensity lesions on diffusion-weighted images in the bilateral frontal and occipital cortices. Myoclonus and periodic sharp-wave complexes on the electroencephalogram were observed in the early disease stage. The clinical diagnosis was typical sCJD. Neuropathologic examination at autopsy showed widespread, characteristic cerebral neocortical involvement with large confluent vacuole-type spongiform change. Spongiform degeneration was also evident in the striatum and medial thalamus. In the cerebellar cortex, slight depletion of Purkinje neurons was evident without spongiform change in the molecular layer or apparent neuron loss in the granule cell layer. The inferior olivary nucleus showed slight hypertrophic astrocytosis without neuron loss. Prion protein (PrP) immunostaining showed widespread, characteristic perivacuolar type PrP deposits with irregular plaque-like PrP deposits in the cerebral neocortex, striatum and medial thalamus. We believe this patient showed early stage cerebral cortical pathology of MM2-cortical-type sCJD, which may provide clues regarding the pathologic progression of this rare sCJD subtype. Although MM2-cortical-type sCJD generally shows slow progression without myoclonus or periodic sharp-wave complexes, the present patient showed a rapidly progressive clinical course similar to that of MM1-type sCJD. PMID- 18410281 TI - CNS lymphomatoid granulomatosis with lymph node and bone marrow involvements. AB - Lymphomatoid granulomatosis (LYG) in the CNS is an uncommon lymphoproliferative disease with characteristic angiocentric lymphoreticular proliferative and granulomatous lesions exhibiting low-grade malignant potential. Here we report a rare case of CNS-LYG, which disseminated to the lymph node and bone marrow. A 50 year-old man was diagnosed with CNS-LYG based on brain biopsy showing perivascular infiltration of CD3-positive small T-lymphocytes without overt nuclear atypism. Eight months after the initial neurological symptoms, inguinal lymph node swelling was found and histopathologically diagnosed as peripheral T cell lymphoma. TCRgamma-gene rearrangement study using both paraffin-embedded specimens of brain and inguinal lymph node demonstrated an identical clonal band. Considering the clinical course, we concluded lymph node involvement of CNS-LYG, suggesting the malignant potential of CNS-LYG. PMID- 18410283 TI - Prevention of paracentesis-induced circulatory dysfunction: midodrine vs albumin. A randomized pilot study. AB - BACKGROUND/AIMS: Large-volume paracentesis in patients with cirrhosis and ascites induces arterial vasodilatation and decreases effective arterial blood volume, termed paracentesis-induced circulatory dysfunction (PICD), which can be prevented by costly intravenous albumin. Vasoconstrictors, e.g. terlipressin, may also prevent PICD. The aim was to compare the less expensive vasoconstrictor midodrine, an alpha-adrenoceptor agonist, with albumin in preventing PICD. METHODS: Twenty-four patients with cirrhosis and ascites were randomly assigned to be treated with either midodrine (n=11) (12.5 mg three times per day; over 2 days) or albumin (n=13) (8 g/L of removed ascites) after large-volume paracentesis. Effective arterial blood volume was assessed indirectly by measuring plasma renin and aldosterone concentration on days 0 and 6 after paracentesis; renal function and haemodynamic changes were also measured. PICD was defined as an increase in plasma renin concentration on day 6 by more than 50% of the baseline value. RESULTS: PICD developed in six patients of the midodrine group (60%) and in only four patients (31%) of the albumin group. Six days after paracentesis, the aldosterone concentration increased significantly in the midodrine group, but not in the albumin group. CONCLUSIONS: This pilot study suggests that midodrine is not as effective as albumin in preventing circulatory dysfunction after large-volume paracentesis in patients with cirrhosis and ascites. PMID- 18410282 TI - Role of hepatic phospholipids in development of liver injury in Mdr2 (Abcb4) knockout mice. AB - BACKGROUND/AIMS: Multidrug resistance protein 2 (Abcb4) gene knockout mice (Mdr2( /-)) lack phosphatidylcholine (PC) excretion into bile and spontaneously develop sclerosing cholangitis, biliary fibrosis and hepatocellular carcinomas. We therefore aimed to test whether formation and hepatic retention of abnormal PC metabolites contribute to the pathogenesis of liver injury in Mdr2(-/-) mice. METHODS: Mdr2(-/-) mice were either fed a diet supplemented with soybean lecithin 2.5% w/w [phosphatidylcholine-enriched diet (PCD), to increase hepatic PC content] or a choline-deficient diet (CDD, to reduce hepatic PC content) for 4 weeks; controls received chow with energy and nutrient content equivalent to PCD and CDD. Serum liver tests, liver histology, markers of fibrosis, cholangiocyte activation, cell proliferation and thin-layer chromatography for phospholipid (PL) composition were carried out. RESULTS: PCD decreased serum alkaline phosphatase and total bilirubin levels compared with controls, while liver histology as well as hepatic hydroxyproline content as markers of liver fibrosis did not differ among groups. Both PCD and CDD decreased hepatocellular proliferation compared with controls. Hepatic, serum and biliary PLs remained unchanged despite dietary manipulations and no potentially toxic PL metabolites were detected. CONCLUSIONS: Mdr2(-/-) mice maintain stable hepatic, serum and biliary PL metabolism in response to dietary PC manipulations. Our findings therefore suggest that liver injury in Mdr2(-/-) mice is not due to formation of toxic PL metabolites. PMID- 18410284 TI - Mitochondrial electron transport inhibition in full genomic hepatitis C virus replicon cells is restored by reducing viral replication. AB - BACKGROUND/AIM: Hepatitis C virus (HCV) core protein has been shown to inhibit mitochondrial electron transport and to increase reactive oxygen species (ROS) in vitro and in vivo. The aim of this study was to investigate whether inhibiting HCV replication could restore the mitochondrial redox state and electron transport activity. METHODS: We measured ROS, mitochondrial reduced glutathione content, and mitochondrial complex I, II, III and IV activities and protein expression in full genomic HCV replicon cells and cured cells that had been prepared by eliminating HCV RNA from replicon cells by interferon (IFN)-alpha treatment. RESULTS: Cured cells had significantly lower ROS production and greater mitochondrial glutathione content than replicon cells. Complete inhibition of HCV replication by IFN-alpha restored complex I and IV activities by 20-30% (P<0.01) and complex I expression (P<0.05). Treatment with fluvastatin, one of the 3-hydroxy-3-methylglutaryl co-enzyme A reductase inhibitors, which is known to have anti-HCV activity, partially inhibited core protein expression and restored complex I activity in full genomic HCV replicon cells to a lesser degree (P<0.05). CONCLUSIONS: Our results show that the mitochondrial redox state and electron transport activity can be restored by reducing HCV replication. PMID- 18410285 TI - kinC/D-mediated heterogeneous expression of spo0A during logarithmical growth in Bacillus subtilis is responsible for partial suppression of phi 29 development. AB - The host of the lytic bacteriophage phi 29 is the spore-forming bacterium Bacillus subtilis. When infection occurs during early stages of sporulation, however, phi 29 development is suppressed and the infecting phage genome becomes trapped into the developing spore. Recently, we have shown that Spo0A, the key transcriptional regulator for entry into sporulation, is directly responsible for suppression of the lytic phi 29 cycle in cells having initiated sporulation. Surprisingly, we found that phi 29 development is suppressed in a subpopulation of logarithmically growing culture and that spo0A is heterogeneously expressed during this growth stage. Furthermore, we showed that kinC and, to a minor extent, kinD, are responsible for heterogeneous expression levels of spo0A during logarithmical growth that are below the threshold to activate sporulation, but sufficient for suppression of the lytic cycle of phi 29. Whereas spo0A was known to be heterogeneously expressed during the early stages of sporulation, our findings show that this also occurs during logarithmical growth. These insights are likely to have important consequences, not only for the life cycle of phi 29, but also for B. subtilis developmental processes. PMID- 18410286 TI - The beta-1,3-glucanosyltransferase gas4p is essential for ascospore wall maturation and spore viability in Schizosaccharomyces pombe. AB - Meiosis is the developmental programme by which sexually reproducing diploid organisms generate haploid gametes. In yeast, meiosis is followed by spore morphogenesis. The formation of the Schizosaccharomyces pombe ascospore wall requires the co-ordinated activity of enzymes involved in the biosynthesis and modification of its components, such as glucans. During sporogenesis, the beta 1,3-glucan synthase bgs2p synthesizes linear beta-1,3-glucans, which remain unorganized and alkali-soluble until covalent linkages are set up between beta 1,3-glucans and other cell wall components. Several proteins belonging to the glycoside hydrolase family 72 (GH72) with beta-1,3-glucanosyltransferase activity have been described in other organisms, such as the Saccharomyces cerevisiae Gas1p or the Aspergillus fumigatus Gel1p. Here we describe the characterization of gas4(+), a new gene that encodes a protein of the GH72 family. Deletion of this gene does not lead to any apparent defect during vegetative growth, but homozygous gas4Delta diploids show a sporulation defect. Although meiosis occurs normally, ascospores are unable to mature or to germinate. The expression of gas4(+) is strongly induced during sporulation and a yellow fluorescent protein (YFP)-gas4p fusion protein localizes to the ascospore periphery during sporulation. We conclude that gas4p is required for ascospore maturation in S. pombe. PMID- 18410287 TI - Population genetics and phylogeography of freshwater mussels in North America, Elliptio dilatata and Actinonaias ligamentina (Bivalvia: Unionidae). AB - Extrinsic and intrinsic forces combined shape the population structure of every species differently. Freshwater mussels are obligate parasites to a host fish during a juvenile stage (glochidia). Elliptio dilatata (ED) and Actinonaias ligamentina (AL) are co-occurring freshwater mussel taxa with similar North American distribution and share some potential host fish. Using mitochondrial DNA, we determined the genotypes of 190 + individuals from collection sites in at least two tributaries in the Lake Erie and Ohio River watersheds, along with the Ouachita and Strawberry rivers in the southeast. Both species had followed a stepping-stone model of dispersal, with greater pairwise genetic structure among collection sites of ED. Also, phylogeographical analysis for ED found significant geographical structuring of haplotype diversity. Overall, within-population variation increased significantly from north to south, with low genetic diversity in the Strawberry River. We calculated significant among-population structure for both species (ED: Phi(ST) = 0.62, P < 0.001; AL: Phi(ST) = 0.16, P < 0.001). Genetic analysis identified the Ouachita River as an area of significant reproductive isolation for both species. Results for AL indicated dispersal into northern areas from two genetically distinct glacial refugia, where results for ED indicated dispersal followed by low gene flow in northern areas. The conservation strategies for mussels that co-occur in the same 'bed' could be species specific. Species such as ED have management units on the population scale, where AL has a more homogeneous genetic structure across its range. PMID- 18410288 TI - Genetic discontinuity, breeding-system change and population history of Arabis alpina in the Italian Peninsula and adjacent Alps. AB - Arabis alpina is a widespread plant of European arctic and alpine environments and belongs to the same family as Arabidopsis thaliana. It grows in all major mountain ranges within the Italian glacial refugia and populations were sampled over a 1300 km transect from Sicily to the Alps. Diversity was studied in nuclear and chloroplast genome markers, combining phylogeographical and population genetic approaches. Alpine populations had significantly lower levels of nuclear genetic variation compared to those in the Italian Peninsula, and this is associated with a pronounced change in within-population inbreeding. Alpine populations were significantly inbred (F(IS) = 0.553), possibly reflecting a change to the self-incompatibility system during leading edge colonization. The Italian Peninsula populations were approaching Hardy-Weinberg equilibrium (outbreeding, F(IS) = 0.076) and genetic variation was highly structured, consistent with independent local 'refugia within refugia' and the fragmentation of an established population by Quaternary climate oscillations. There is very little evidence of genetic exchange between the Alps and the Italian Peninsula main distribution ranges. The Alps functioned as a glacial sink for A. alpina, while the Italian Peninsula remains a distinct and separate long-term refugium. Comparative analysis indicated that inbreeding populations probably recolonized the Alps twice: (i) during a recent postglacial colonization of the western Alps from a Maritime Alps refugium; and (ii) separately into the central Alps from a source outside the sampling range. The pronounced geographical structure and inbreeding discontinuities are significant for the future development of A. alpina as a model species. PMID- 18410289 TI - Geographical variation of genetic and phenotypic traits in the Mexican sailfin mollies, Poecilia velifera and P. petenensis. AB - Comparing the patterns of population divergence using both neutral genetic and phenotypic traits provides an opportunity to examine the relative importance of evolutionary mechanisms in shaping population differences. We used microsatellite markers to examine population genetic structure in the Mexican sailfin mollies Poecilia velifera and P. petenensis. We compared patterns of genetic structure and divergence to that in two types of phenotypic traits: morphological characters and mating behaviours. Populations within each species were genetically distinct, and conformed to a model of isolation by distance, with populations within different geographical regions being more genetically similar to one another than were populations from different regions. Bayesian clustering and barrier analyses provided additional support for population separation, especially between geographical regions. In contrast, none of the phenotypic traits showed any type of geographical pattern, and population divergence in these traits was uncorrelated with that found in neutral markers. There was also a weaker pattern of regional differences among geographical regions compared to neutral genetic divergence. These results suggest that while divergence in neutral traits is likely a product of population history and genetic drift, phenotypic divergence is governed by different mechanisms, such as natural and sexual selection, and arises at spatial scales independent from those of neutral markers. PMID- 18410290 TI - Factors affecting the reproductive success of dominant male meerkats. AB - Identifying traits that affect the reproductive success of individuals is fundamental for our understanding of evolutionary processes. In cooperative breeders, a dominant male typically restricts mating access to the dominant female for extended periods, resulting in pronounced variation in reproductive success among males. This may result in strong selection for traits that increase the likelihood of dominance acquisition, dominance retention and reproductive rates while dominant. However, despite considerable research on reproductive skew, few studies have explored the factors that influence these three processes among males in cooperative species. Here we use genetic, behavioural and demographic data to investigate the factors affecting reproductive success in dominant male meerkats (Suricata suricatta). Our data show that dominant males sire the majority of all offspring surviving to 1 year. A male's likelihood of becoming dominant is strongly influenced by age, but not by weight. Tenure length and reproductive rate, both important components of dominant male reproductive success, are largely affected by group size and composition, rather than individual traits. Dominant males in large groups have longer tenures, but after this effect is controlled, male tenure length also correlates negatively to the number of adult females in the group. Male reproductive rate also declines as the number of intra- and extra-group competitors increases. As the time spent in the dominant position and reproductive rate while dominant explain > 80% of the total variance in reproductive success, group composition thus has major implications for male reproductive success. PMID- 18410291 TI - Origin and genetic diversity of Western European populations of the potato cyst nematode (Globodera pallida) inferred from mitochondrial sequences and microsatellite loci. AB - Native to South America, the potato cyst nematode Globodera pallida is one of the principal pests of Andean potato crops and is also an important global pest following its introduction to Europe, Africa, North America, Asia and Oceania. Building on earlier work showing a clear south to north phylogeographic pattern in Peruvian populations, we have been able to identify the origin of Western European populations with high accuracy. They are all derived from a single restricted area in the extreme south of Peru, located between the north shore of the Lake Titicaca and Cusco. Only four cytochrome b haplotypes are found in Western Europe, one of them being also found in some populations of this area of southern Peru. The allelic richness at seven microsatellite loci observed in the Western European populations, although only one-third of that observed in this part of southern Peru, is comparable to the allelic richness observed in the northern region of Peru. This result could be explained by the fact that most of the genetic variability observed at the scale of a field or even of a region is already observed at the scale of a single plant within a field. Thus, even introduction via a single infected potato plant could result in the relatively high genetic variability observed in Western Europe. This finding has important consequences for the control of this pest and the development of quarantine measures. PMID- 18410292 TI - Recent divergence with gene flow in Tennessee cave salamanders (Plethodontidae: Gyrinophilus) inferred from gene genealogies. AB - Cave organisms occupy a special place in evolutionary biology because convergent morphologies of many species demonstrate repeatability in evolution even as they obscure phylogenetic relationships. The origin of specialized cave-dwelling species also raises the issue of the relative importance of isolation vs. natural selection in speciation. Two alternative hypotheses describe the origin of subterranean species. The 'climate-relict' model proposes allopatric speciation after populations of cold-adapted species become stranded in caves due to climate change. The 'adaptive-shift' model proposes parapatric speciation driven by divergent selection between subterranean and surface habitats. Our study of the Tennessee cave salamander complex shows that the three nominal forms (Gyrinophilus palleucus palleucus, G. p. necturoides, and G. gulolineatus) arose recently and are genealogically nested within the epigean (surface-dwelling) species, G. porphyriticus. Short branch lengths and discordant gene trees were consistent with a complex history involving gene flow between diverging forms. Results of coalescent-based analysis of the distribution of haplotypes among groups reject the allopatric speciation model and support continuous or recurrent genetic exchange during divergence. These results strongly favour the hypothesis that Tennessee cave salamanders originated from spring salamanders via divergence with gene flow. PMID- 18410293 TI - Ancient DNA and conservation: lessons from the endangered kiwi of New Zealand. AB - Conservation genetics typically seeks to map the distribution of contemporary genetic variation across space, and to use the resulting genetic parameters to infer any likely short-term evolutionary consequences for rare and endangered species. Recent developments in the study of ancient DNA now enable the extension of genetic variation studies backwards in time and provide a context by which to interpret contemporary levels of genetic variation, in addition to any patterns of genetic change over time. Ancient DNA research can also help to determine past levels of genetic diversity, identify species' boundaries and reveal former ranges among morphologically cryptic taxa. Ancient DNA sequence data for the New Zealand kiwi (Apteryx spp.) are presented and we show that most ancient populations of rowi and tokoeka exhibited private mitochondrial DNA haplotypes. Moreover, we illustrate that the extinction of these populations has led to the loss of considerable genetic variation. We also use ancient DNA methodology to determine past species distributions for brown kiwi and great spotted kiwi whose bones are morphologically indistinguishable. PMID- 18410294 TI - Population genetic structure across dissolved oxygen regimes in an African cichlid fish. AB - Ecological isolation is a process whereby gene flow between selective environments is reduced due to selection against maladapted dispersers, migrant alleles, or hybrids. Although ecological isolation has been documented in several systems, gene flow can often be high among selective regimes, and more studies are thus needed to better understand the conditions under which ecological gradients or divergent selective regimes should influence population structure. We test for ecological isolation in a system in which high plasticity occurs with respect to traits that are adaptive in alternate forms under different environmental conditions. Pseudocrenilabrus multicolor victoriae is a widespread haplochromine cichlid fish in East Africa that exploits both normoxic (normal oxygen) rivers/lakes and hypoxic (low oxygen) swamps. Here, we examine population structure, using mitochondrial DNA and microsatellites, to determine if genetic divergence is significantly increased between dissolved oxygen regimes relative to within them, while controlling for geographical structure. Our results indicate that geographical separation influences population structure, while no effects of divergent selection with respect to oxygen regimes were detected. Specifically, we document (i) genetic clustering according to geographical region, but no clustering according to oxygen regime; (ii) higher genetic variation among than within regions, but no effect of oxygen regime on genetic variation; (iii) isolation by distance within one region; and (iv) decreasing genetic variability with increasing geographical distance from Lake Victoria. We speculate that plasticity may be facilitating gene flow between oxygen regimes in this system. PMID- 18410295 TI - Speciation with gene flow could be common. AB - The likelihood of speciation in the face of homogenizing gene flow (i.e. without complete geographical isolation) is one of the most debated topics in evolutionary biology. Demonstrating this phenonemon is hampered by the difficulty of isolating the effects of time since population divergence vs. gene flow on levels of molecular genetic differentiation. For example, weak genetic differentiation between taxa could be due to recent divergence, gene flow, or a combination of these factors. Nonetheless, a number of convincing examples of speciation with gene flow have recently emerged, owing in part to the development of new analytical methods designed to estimate gene flow specifically. A recent example of speciation with gene flow in salamanders (Niemiller et al. 2008) further advances our understanding of this phenonemon, by showing that gene flow between cave and spring salamanders was ongoing during speciation, rather than having occurred after a long period of allopatric divergence. Future work on the ecological and genetic factors reducing gene flow will likely increase our understanding of the conditions that faciliate divergence in the face of gene flow. PMID- 18410296 TI - Two challenges for the classification of sexual dysfunction. AB - INTRODUCTION: The current classification of sexual function (in particular, the Diagnostic and Statistical Manual of Mental Disorders, DSM-IV) has lately attracted significant criticism at both research and clinical levels. Despite this, there has been a reluctance to return to the drawing board. Instead, attempts to improve the system have been marginal, constrained by the need to secure professional consensus, the desire for continuity with traditional categories, and the emphasis on diagnostic agreement (reliability). AIM: In this article, we examine two key challenges currently faced by the DSM: how to effectively acknowledge the relational context of sexual problems and how to avoid pathologizing normal variation. RESULTS: We raise some possible new directions, such as ways in which relational processes could be integrated into the current system, and possibilities for introducing a dimensional rather than a categorical model of sexual function. CONCLUSIONS: We argue that if the next version of DSM (version V) is to avoid the weaknesses inherent in the present system, then a return to the drawing board is precisely what is required. PMID- 18410297 TI - The effects of citalopram and fluoxetine on sexual behavior in healthy men: evidence of delayed ejaculation and unaffected sexual desire. A randomized, placebo-controlled, double-blind, double-dummy, parallel group study. AB - INTRODUCTION: Selective serotonin reuptake inhibitors (SSRIs) are known to induce delayed orgasm and delayed ejaculation, while their effect on other aspects of sexual function, such as sexual motivation, arousal, and erectile function are unclear. AIM: In order to evaluate the effect of chronic administration of two SSRIs, citalopram and fluoxetine, on normal sexual function, we studied the parameters of male sexual behavior, erectile function, and ejaculation on 48 healthy male volunteers, aged 29.5 +/- 4.9, in a randomized, placebo-controlled, double-blind, double-dummy study. Methods. The subjects were randomized to receive placebo (16 subjects), or fluoxetine (20 mg/day) (16 subjects) or citalopram (20 mg/day) for the first week, and 40 mg/day in the following 3 weeks (16 subjects). MAIN OUTCOME MEASURES: Sexual function was investigated at the screening and at the end of the study by means of test of penile erection (TPE) and masturbation ejaculation latency time (MELT) performed during visual erotic stimulation, and at each visit by self-filled questionnaires (International Index Erectile Function [IIEF-15] and Golombock Rust Inventory of Sexual Satisfaction [GRISS]). RESULTS: All the erectile parameters, evaluated by means of RigiScan Plus during TPE, were not significantly different when both fluoxetine and citalopram were compared with placebo. A delay in the ejaculation time was observed both during citalopram and during fluoxetine treatment when compared with placebo, reaching a statistical significance only with citalopram. During the treatment with citalopram and fluoxetine, the IIEF-15 score of all items decreased except for those items related to sexual desire; however, the scores were significantly lower only for the citalopram treatment. CONCLUSIONS: The treatment with citalopram or with fluoxetine was confirmed to delay ejaculation, but was significant only for citalopram. Citalopram and fluoxetine did not affect sexual desire. Citalopram and fluoxetine did not directly affect penile erection as objectively assessed by RigiScan, although an impairment in the subjective assessment of erectile function was observed, but was significant only for citalopram, and it was thought to be a possible consequence of the delayed ejaculation perceived as a trouble. PMID- 18410299 TI - Magnetic resonance imaging and the female sexual response: overview of techniques, results, and future directions. AB - INTRODUCTION: Magnetic resonance imaging (MRI) is becoming a frequently used tool in the study of sexual physiology and neurophysiology. AIM: This report reviews various MRI methods used to study the female sexual arousal response. METHODS: Retrospective review of pertinent literature. RESULTS: Dynamic genital MRI studies enable the visualization of the physiologic arousal response that provides the direct observation of the time course and magnitude of this response, along with the variability that appears to occur in women with sexual arousal disorder. Functional brain MR studies are described and summarized along with an overview of what we have learned. Finally, the speculation on how we may be able to use MRI technology to better understand the female sexual response and to help in validating new drug treatments or in devising new treatment strategies for sexual dysfunction is also presented. CONCLUSIONS: Neuroimaging has already been proven as an invaluable research tool to study the sexual response in women both in the pelvis as well as within the brain. Using these techniques, major inroads are being made to improve the understanding of the sexual arousal process in women. PMID- 18410298 TI - Sexual satisfaction among patients with erectile dysfunction treated with counseling, sildenafil, or both. AB - INTRODUCTION: Sexual satisfaction is linked to life satisfaction, and erectile dysfunction (ED) may lead to an impaired quality of life (QOL). AIM: Our goal was to evaluate the QOL among Brazilian patients with ED, before and after three kinds of treatment. METHODS: Men aged 25-55 years, with a diagnosis of psychogenic or mixed ED, according to the Classification of Mental and Behavioral Disorders of the International Classification of Diseases, 10th edition, and the Standard Practice in Sexual Medicine, were randomly assigned to three treatment groups: counseling, sildenafil, and sildenafil plus counseling. At baseline each group had 40 patients. Sildenafil was provided in 50 mg that could be adjusted to 100 mg. The patients could initially take one to two tablets per week and the entire treatment lasted for 3 months. Counseling was provided in group sessions that took place once a week. They were evaluated at baseline and after 3 months of treatment with the Male Sexual Quotient (MSQ) and the Sexual Health Inventory for Men (SHIM). MAIN OUTCOME MEASURES: The correlation between the patients' MSQ score and scores on the SHIM. RESULTS: One hundred seventeen patients were enrolled. The three groups were similar according to age, marital status, mean time of ED, and ED severity and etiology. At baseline, MSQ and SHIM total scores were not different among the three groups. MSQ scores increased from 41.2 +/- 15.3, 38.7 +/- 18.0, and 46.8 +/- 17.0 to 48.5 +/- 15.3, 63.8 +/- 21.6, and 70.0 +/- 17.3 after counseling, sildenafil, and sildenafil plus counseling, respectively (P < 0.05). SHIM scores also increased significantly (9.6 +/- 4.1, 9.7 +/- 4.1, and 10.2 +/- 3.9 to 12.1 +/- 3.9, 16.7 +/- 5.6, and 17.7 +/- 4.5 after counseling, sildenafil, and sildenafil plus counseling, respectively) (P < 0.05). There were no serious adverse events related to sildenafil, and no patient was withdrawn from the study because of an adverse event. CONCLUSIONS: The three treatments were significantly efficient, and the best treatment was sildenafil associated with counseling. PMID- 18410300 TI - Risk factors for female sexual dysfunction in the general population: exploring factors associated with low sexual function and sexual distress. AB - INTRODUCTION: No previous population-based studies have used validated instruments to measure female sexual dysfunction (FSD) in Australian women across a broad age range. AIM: To estimate prevalence and explore factors associated with the components of FSD. MAIN OUTCOME MEASURES: Sexual Function Questionnaire measured low sexual function. Female Sexual Distress Scale measured sexual distress. Methods. Multivariate analysis of postal survey data from a random sample of 356 women aged 20-70 years. RESULTS: Low desire was more likely to occur in women in relationships for 20-29 years (odds ratio 3.7, 95% confidence intervals 1.1-12.8) and less likely in women reporting greater satisfaction with their partner as a lover (0.3, 0.1-0.9) or who placed greater importance on sex (0.1, 0.03-0.3). Low genital arousal was more likely among women who were perimenopausal (4.4, 1.2-15.7), postmenopausal (5.3, 1.6-17.7), or depressed (2.5, 1.1-5.3), and was less likely in women taking hormone therapy (0.2, 0.04 0.7), more educated (0.5, 0.3-0.96), in their 30s (0.2, 0.1-0.7) or 40s (0.2, 0.1 0.7), or placed greater importance on sex (0.2, 0.05-0.5). Low orgasmic function was less likely in women who were in their 30s (0.3, 0.1-0.8) or who placed greater importance on sex (0.3, 0.1-0.7). Sexual distress was positively associated with depression (3.1, 1.2-7.8) and was inversely associated with better communication of sexual needs (0.2, 0.05-0.5). Results were adjusted for other covariates including age, psychological, socioeconomic, physiological, and relationship factors. CONCLUSIONS: Relationship factors were more important to low desire than age or menopause, whereas physiological and psychological factors were more important to low genital arousal and low orgasmic function than relationship factors. Sexual distress was associated with both psychological and relationship factors. PMID- 18410301 TI - Genital responsiveness in healthy women with and without sexual arousal disorder. AB - INTRODUCTION: Most pharmacological treatments that are currently being developed for women with sexual arousal disorder are aimed at remedying a vasculogenic deficit. AIM: This study investigated whether pre- and postmenopausal women with sexual arousal disorder are less genitally responsive to visual sexual stimuli than pre- and postmenopausal women without sexual problems. METHOD: Twenty-nine medically healthy women with sexual arousal disorder (15 premenopausal and 14 postmenopausal), diagnosed using the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition (DSM-IV) criteria, and 30 age-matched women without sexual problems (16 premenopausal and 14 postmenopausal) were shown sexual stimuli depicting cunnilingus and intercourse. MAIN OUTCOME MEASURE: Genital arousal was assessed as vaginal pulse amplitude (VPA) using vaginal photoplethysmography. RESULTS: Results showed no significant differences between the two groups in mean and maximum VPA, nor in latency of VPA response. CONCLUSION: Women with sexual arousal disorder diagnosed according to DSM-IV criteria were not less genitally responsive to visual sexual stimuli than women without such problems. These findings are in line with previous studies. The sexual problems these women report are clearly not related to their potential to become genitally aroused. We argue that the DSM-IV criteria for sexual arousal disorder are in need of revision. In medically healthy women, impaired genital responsiveness is not a valid diagnostic criterion. PMID- 18410303 TI - After 15 years, has the time come to revise the term "erectile dysfunction?". PMID- 18410302 TI - Exploring sexual attitudes of students in health professions. AB - INTRODUCTION: Patients' sexual concerns are frequently underestimated because of health professionals' reluctance to address sexual health issues. Though it has been documented that sexual attitudes are extremely influential in everyday clinical practice, limited data exist on identifying health professionals' attitudes. AIM: To explore sexual attitudes in medical students and students in allied health professions. MAIN OUTCOME MEASURES: The Derogatis Sexual Functioning Inventory (DSFI)-Attitude Subscale was used to assess sexual attitudes. METHODS: The sample included 1st and 4th year college students enrolled in the following academic departments/schools of two academic institutions leading to health professions: medical school, psychology, pharmaceutical school, nursing and midwifery. Demographic data were obtained relating to sexual behaviors and information sources on sexual issues. Data were analyzed using independent samples t-test and two-way analysis of variance, as well as logistic regression and Pearson's correlation coefficient. RESULTS: A total of 714 students (81.9% females) participated in the study: 48.5% 1st year students and 51.5% seniors with a mean age of 20.17 years (SD = 1.87, range 17 25). Using iterative cluster analysis on DSFI scores, participants were divided in conservative (N = 167), liberal (N = 224), and neutral (N = 323) clusters. A significant gender difference on sexual attitudes was obtained (P < 0.001) with male students being more liberal compared to females (mean = 18.26 and mean = 11.13, respectively). Differences were also revealed for the field but not for the year of study. Analysis also revealed that liberalism in sexual attitudes is more likely to be affected by a liberal stance toward religion (OR: 2.39), receiving information for sexual matters mainly from peers (OR: 1.86), and media influence on students' sexual life (OR: 1.68). CONCLUSIONS: Gender, personal values, and experiences influence students' attitudes toward sexual issues. Since negative attitudes can impede effective sexual health consultations, it is imperative to incorporate courses on effective communication and human sexuality in the medical and allied health professions curricula that will enhance students' awareness of their own values and prejudices. PMID- 18410304 TI - Phenotypic assessment of endothelial microparticles in diabetic and nondiabetic men with erectile dysfunction. AB - INTRODUCTION: Endothelial microparticles (EMPs) may play a role as biomarkers of vascular injury. EMPs are higher in men with diabetes diabetic men with erectile dysfunction (ED) than in nondiabetic potent men. AIM: The aim of this study was to quantize different phenotypic circulating EMP levels among diabetic and nondiabetic patients with ED, and to determine whether EMPs are released as a result of activation or apoptosis. METHODS: We studied 30 type 2 diabetic and 24 nondiabetic subjects with symptomatic ED from at least 6 months, and 20 nondiabetic men without ED matched for age and weight with diabetic and nondiabetic subjects. Erectile function was assessed by completing the International Index of Erectile Function (IEEF)-5, which consists of Items 5, 15, 4, 2, and 7 from the full-scale IIEF-15. A score of 21 or less indicates the presence of ED. MAIN OUTCOME MEASURES: EMP levels in plasma were quantified by flow cytometry. Markers for apoptosis (platelet/endothelial cell adhesion molecule 1/CD31 antigen) and activation (E-selectin/CD62E antigen) were compared. Endothelium-dependent flow-mediated dilation (FMD) was evaluated in the right brachial artery with a high-resolution ultrasound machine following reactive hyperemia. RESULTS: Diabetic patients were found to have the highest levels of EMP31+; diabetic and nondiabetic men with ED were found to have significantly higher levels of EMP62+ than nondiabetic men without ED. The EMP62/EMP31 ratio, an index of endothelial activation (high ratio) or apoptosis (low ratio), was lowest in diabetic men with ED (0.20). In the whole group of 54 men with ED (diabetic and nondiabetic), there was an inverse correlation between FMD and the number of circulating EMPs (P < 0.05). CONCLUSIONS: The presence of diabetes in subjects with ED is associated with a different pattern of endothelial cell injury. The phenotypic assessment of EMPs in diabetic patients with ED is consistent with increased apoptotic activity. PMID- 18410305 TI - Socioeconomic status, not race/ethnicity, contributes to variation in the prevalence of erectile dysfunction: results from the Boston Area Community Health (BACH) Survey. AB - INTRODUCTION: Few population-based studies have assessed variations in the burden of erectile dysfunction (ED) by race/ethnicity. AIM: To estimate prevalence rates of ED by race/ethnicity and determine the contribution of behavioral risk factors, chronic illnesses, and socioeconomic factors to potential race/ethnic differences in ED. METHODS: The Boston Area Community Health (BACH) Survey is a study of urologic symptoms in a racially and ethnically diverse population. BACH used a multistage stratified random sample to recruit 2,301 men aged 30-79 years from the city of Boston. Self-reported race/ethnicity was defined as Black, Hispanic, and White. Socioeconomic status (SES) was defined as a combination of education and household income. MAIN OUTCOME MEASURES: ED assessed using the 5 item International Index of Erectile Function (IIEF-5). IIEF-5 scores were used both as a continuous variable and dichotomized as /=17. RESULTS: Overall prevalence of ED (defined as an IIEF-5 score or= 3 months on or= 3 months with 256 microg ml(-1)) and trimethoprim (MIC of >32 microg ml(-1)); few were resistant to clindamycin (MIC(90) 4 microg ml(-1)), erythromycin (MIC(90) 8 microg ml(-1)) and tetracycline (MIC(90) 8 microg ml(-1)); but none was resistant to azithromycin (MIC(90) 4 microg ml(-1)), ciprofloxacin (MIC(90) 1 microg ml(-1)) or gentamycin (MIC(90) 4 microg ml(-1)). Most strains restricted with SmaI, but a combination of SmaI-KpnI digestion was more discriminatory. MLST analysis yielded four sequence types (ST), and ST-2624 was the predominant one. Phylogenetic analysis revealed a high degree of recombination for glnA and pgm genes. The predominant serotypes were O:3 and O:5. Most strains had lowest cytotoxicity potential with Caco-2 cells, medium cytotoxicity with INT-407 and Hep-2 cells and high cytotoxicity with CHO cells. CONCLUSION: A low degree of antimicrobial resistance, 13 PFGE profiles, 4 ST and a large variability in cytotoxicity assays were found for these strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first characterization of C. jejuni strains isolated from broilers in Puerto Rico. The genetic diversity of these strains suggests that several techniques are needed for strain characterization. PMID- 18410343 TI - Characterization of an Helicobacter pylori environmental strain. AB - AIMS: To investigate the main genotypic virulence markers and the phenotypic features of an environmental Helicobacter pylori strain, named MDC1. METHODS AND RESULTS: The H. pylori MDC1 genotypic status was evaluated by PCR amplification. The mosaicism in vacA alleles was expressed by the s1m1 allelic combination, as found in strains which are strong vacuolating cytotoxin producers; the number of cagA variable EPIYA motifs displayed P1P2P3P3 pattern and the iceA1 was recorded between the iceA allelic types and the babA2 gene found in strains causing more severe disease. The biofilm formation was evaluated on a polystyrene surface in static conditions by scanning electron microscopy and confocal scanning laser microscopy. Helicobacter pylori MDC1 displayed a dense mature biofilm with cells in a coccoid morphology persistent in time in which the expression of the luxS gene, related to the quorum-sensing signalling, was always detected. CONCLUSIONS: Helicobacter pylori MDC1 strain had the main virulence markers closely related to gastric pathogenesis and displayed a well-structured biofilm which allowed this bacterium to be more protected in the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The persistence of the environmental virulent H. pylori strain in a clustered state suggests a long-term survival of this bacterial community outside of the host, enabling the bacterial transmission with important clinical repercussions. PMID- 18410344 TI - Investigation into the effect of detergents on disinfectant susceptibility of attached Escherichia coli and Listeria monocytogenes. AB - AIMS: Investigate the effect of detergent treatment on susceptibility of attached Escherichia coli and Listeria monocytogenes to subsequent disinfectant treatment. METHODS AND RESULTS: Plate counts show that E. coli attached to stainless steel surfaces became significantly more susceptible to benzalkonium chloride (BAC) after treatment with sodium alkyl sulfate (SAS) and fatty alcohol ethoxylate (FAE). No change in susceptibility was observed with Sodium dodecyl sulfate (SDS). L. monocytogenes became significantly less susceptible to BAC after treatment with SAS and SDS yet no change in susceptibility was observed with FAE. Flow cytometry using the fluoresceine propidium iodide revealed significant increases in cell membrane permeability of both organisms by SAS and FAE, although the effect was much greater in E. coli. No change was observed with SDS. Hydrophobic interaction chromatography showed that both organisms became less hydrophobic following treatment with SAS and SDS but FAE had no effect. CONCLUSIONS: In E. coli, detergents that increase susceptibility to BAC increase membrane permeability. In L. monocytogenes, detergents that reduce susceptibility to BAC lower cell surface hydrophobicity. SIGNIFICANCE AND IMPACT OF THE STUDY: Detergents can influence the sensitivity of pathogenic food borne micro-organisms to BAC. PMID- 18410345 TI - Characterization of Sro1, a novel stress responsive protein in Schizosaccharomyces pombe. AB - The large amount of available genome sequencing data presents a huge challenge in the form of orphan sequences. This study reports the detailed functional characterization of one such orphan sequence in Schizosaccharomyces pombe. We identified this gene as a prominently upregulated 1.4 kb transcript in a screen for Cigarette smoke extract responsive genes in S. pombe and named it Stress Responsive Orphan 1 (Sro1). We report various functions of Sro1 in regulation of cellular behaviour under stress conditions. We show that this gene (Sro1) responds to a variety of stress conditions and that the expression of the gene is regulated mainly through the stress activated protein kinase (SAPK) Sty1 and its downstream transcription factor Atf1. Deletion of Sro1 also significantly alters the reactive oxygen species (ROS) generation profiles and the cell-cycle progression of S. pombe during stress conditions. The stress-specific alteration of the ROS generation profiles and checkpoint activation resulting from deletion of the gene suggest that Sro1 might be a key player in determining cellular responses/fate under stress conditions. PMID- 18410346 TI - Characterization of the recombinant diaminobutyric acid acetyltransferase from Methylophaga thalassica and Methylophaga alcalica. AB - Diaminobutyric acid acetyltransferase (EctA) catalyzes the acetylation of diaminobutyric acid to gamma-N-acetyl-alpha,gamma-diaminobutyrate with acetyl coenzyme A. This is the second reaction in the ectoine biosynthetic pathway. The recombinant EctA proteins were purified from two moderately halophilic methylotrophic bacteria: Methylophaga thalassica ATCC 33146T and Methylophaga alcalica ATCC 35842T. EctA found in both methylotrophs is a homodimer with a subunit molecular mass of c. 20 kDa and had similar properties with respect to the optimum temperature for activity (30 degrees C), Km for diaminobutyrate (370 or 375 microM) and the absence of requirements for divalent metal ions. The enzyme from M. thalassica exhibited a lower pH optimum and was inhibited both by sodium carbonates and by high ionic strength but to a lesser extent by copper ions. PMID- 18410347 TI - Phosphate and sulfur limitation responses in the chloroplast of Chlamydomonas reinhardtii. AB - Phosphorus (P) and sulfur (S) are two macronutrients that photosynthetic organisms require in relatively large amounts despite their levels in the environment often being limited. Accordingly, to adapt to random changes in macronutrient concentrations, plants and algae must sense and respond in a coordinated fashion. The unicellular green alga Chlamydomonas reinhardti is a widely used model organism for the study of P and S stress responses. Herein, we review the current knowledge of P and S nutrient stress responses, highlighting the roles of P and S key global-regulator proteins in mediating signals that link P and S detection to different chloroplast nutrient stress responses. PMID- 18410348 TI - Intravenous iron attenuates postvaccination anti-HBsAg titres after quadruple hepatitis B vaccination in dialysis patients with erythropoietin therapy. AB - BACKGROUND: Anaemia in patients with end-stage renal disease (ESRD) is commonly treated with recombinant human erythropoietin (rHuEPO), often in combination with an adjuvant iron supplement. There is much evidence that rHuEPO can influence the immune response by its effect on lymphocytes. Also, iron catalyses the formation of radicals and increases the risk of major infections by negatively affecting the immune system. The relationship between antibodies to hepatitis B surface antigen (anti-HBsAg) responsiveness after hepatitis B vaccination and rHuEPO/adjuvant iron supplementation has not been reported before. AIM: To determine the effects of subcutaneous erythropoietin and intravenous (i.v.) iron therapy on the responsiveness of anti-HBsAg after quadruple hepatitis B vaccination among ESRD patients. METHODS: Retrospective medical records were reviewed in a hospital with a tertiary teaching facility. Eighty-three ESRD patients, including 51 who underwent haemodialysis and 32 who underwent peritoneal dialysis therapy, received a quadruple recombinant hepatitis B vaccine. We investigated anti-HBsAg titres in those patients who either received rHuEPO alone (n = 50) or rHuEPO in combination with i.v. iron (n = 33). RESULTS: We found that the postvaccination anti-HBsAg titre was significantly lower in the rHuEPO plus i.v. iron group when compared with the group with rHuEPO alone (p < 0.05). The increment of anti-HBsAg between the initial month and the seventh month was positively correlated with therapeutic rHuEPO dosages in the group with rHuEPO alone (r = 0.303, p = 0.033). This relationship was not present in the rHuEPO with i.v. iron group (r = -0.289, p = 0.229). CONCLUSIONS: The levels of anti-HBsAg after hepatitis B vaccination are positively correlated with the dose of rHuEPO treatment during the vaccinated period among ESRD patients without i.v. iron supplementation. Also, i.v. iron negatively impacts the responsiveness of anti-HBsAg titre after hepatitis B vaccination in ESRD patients who have undergone rHuEPO therapy. PMID- 18410349 TI - The effect of parental consanguinity on the clinical and laboratory findings of rheumatoid arthritis. AB - AIMS: We aimed to evaluate the frequency of consanguinity among the parents of patients with rheumatoid arthritis (RA) and the influence of parental consanguinity on several clinical and laboratory parameters which reflect the severity of the disease. METHODS AND PATIENTS: The study population consisted of 265 patients with RA which were divided into two groups with respect to the presence or absence of consanguinity between their parents. The frequency of parental consanguinity was compared with the general population. The two groups were compared with respect to family history of RA, the age of onset, the age at which RA was diagnosed, duration of the disease, the presence of rheumatoid nodules, vasculitis, serositis and the need for orthopaedic surgery, amyloidosis, the presence and level of rheumatoid factor and anti-cyclic citrullinated peptide antibodies, erosive changes on radiographs, and the need for anti-tumour necrosis factor therapy. RESULTS: Twenty-one patients (8%) had parents who were consanguineous, which was not more frequent compared with the general population (14%). The mean age of disease onset and the mean age at which RA was diagnosed were lower in patients with parental consanguinity, although the difference was not statistically significant. The other clinical and laboratory parameters were also not different between the two groups. CONCLUSION: The present data suggests that parental consanguinity has no effect on disease severity, and the frequency of consanguinity is not increased among the parents of patients with RA. A possible exception is the earlier disease onset and age at diagnosis which needs to be confirmed by larger studies. PMID- 18410350 TI - Measuring flushing symptoms with extended-release niacin using the flushing symptom questionnaire: results from a randomised placebo-controlled clinical trial. AB - INTRODUCTION: Niacin is underutilised because of flushing. Lack of a quantitative tool to assess niacin-induced flushing has precluded the objective evaluation of flushing associated with extended-release (ER) niacin formulations. We developed the Flushing Symptom Questionnaire((c)) (FSQ), a quantitative tool to assess patient-reported flushing, and assessed its ability to characterise ER niacin induced flushing. METHODS: This study focused on the responses to one question in the FSQ, the Global Flushing Severity Score (GFSS), reported on a 0-10 scale (none = 0, mild = 1-3, moderate = 4-6, severe = 7-9 and extreme = 10) to assess flushing during ER niacin initiation (week 1) and maintenance (weeks 2-8). RESULTS: Flushing severity with ER niacin was greatest during week 1 and remained greater than placebo for the study duration. During weeks 2-8, 40% of patients on ER niacin vs. 8% of those on placebo had > 1 day/week with 'moderate or greater' GFSS. CONCLUSIONS: In conclusion, the GFSS component of the FSQ was a sensitive and responsive quantitative measure of ER niacin-induced flushing that will aid in the objective comparison of novel strategies intended to improve tolerability and adherence to niacin, an agent proven to reduce cardiovascular risk. PMID- 18410351 TI - Clinical characteristics of patients with diabetic polyneuropathy: the role of clinical and electromyographic evaluation and the effect of the various types on the quality of life. AB - OBJECTIVE: This study was performed to identify the relationship between the quality of life and polyneuropathy which is one of the complications of diabetes. METHODS: Total 111 patients with diabetes mellitus were taken into the study as type 1 and type 2. Patients were accepted having polyneuropathy according to their electroneuromyography (ENMG) results. To evaluate the quality of life in the patients Short Form 36 (SF-36) and World Health Organization Quality of Life Questionnaire abbreviated version (WHOQOL-BREF) were used. RESULTS: Clinical polyneuropathy was found in 46% of the patients, while polineuropathy was found in 63% of the patients with evaluation ENMG. The patients with polyneuropathy had poor quality of life according to SF-36 and WHOQOL-BREF (p < 0.001). The mean quality of life scores of patients who had sensoriomotor and mix polyneuropathy, were lower than sensory type and axonal polyneuropathy. CONCLUSION: Diabetic polyneuropathy influences the quality of life in a negative way. The quality of life scores of patients who had polyneuropathy continuing with mixed pathogenesis and sensoriomotor type, become worse for this reason, even if the patients do not have any clinical polyneuropathy, this being evaluated with ENMG. PMID- 18410352 TI - Effects of reiki in clinical practice: a systematic review of randomised clinical trials. AB - INTRODUCTION: The aim of this systematic review is to summarise and critically evaluate the evidence for the effectiveness of reiki. METHODS: We searched the literature using 23 databases from their respective inceptions through to November 2007 (search again 23 January 2008) without language restrictions. Methodological quality was assessed using the Jadad score. RESULTS: The searches identified 205 potentially relevant studies. Nine randomised clinical trials (RCTs) met our inclusion criteria. Two RCTs suggested beneficial effects of reiki compared with sham control on depression, while one RCT did not report intergroup differences. For pain and anxiety, one RCT showed intergroup differences compared with sham control. For stress and hopelessness a further RCT reported effects of reiki and distant reiki compared with distant sham control. For functional recovery after ischaemic stroke there were no intergroup differences compared with sham. There was also no difference for anxiety between groups of pregnant women undergoing amniocentesis. For diabetic neuropathy there were no effects of reiki on pain. A further RCT failed to show the effects of reiki for anxiety and depression in women undergoing breast biopsy compared with conventional care. DISCUSSION: In total, the trial data for any one condition are scarce and independent replications are not available for each condition. Most trials suffered from methodological flaws such as small sample size, inadequate study design and poor reporting. CONCLUSION: In conclusion, the evidence is insufficient to suggest that reiki is an effective treatment for any condition. Therefore the value of reiki remains unproven. PMID- 18410353 TI - Tuberculosis-HIV co-infection: policy and epidemiology in 25 countries in the WHO European region. AB - OBJECTIVES: The aims of this study were to collect and review tuberculosis (TB) HIV data for Europe and to provide an overview of current health policies addressing co-infection. METHODS: We collected reported cases of TB-HIV from the 25 most affected member states of the World Health Organization (WHO) European region. Countries were also asked whether they had implemented health policies covering collaborative TB-HIV activities and what their main achievements, obstacles and needs were in addressing TB-HIV. RESULTS: Twenty countries reported registering a total of 6925 TB-HIV cases in 2005. Among TB patients tested, 3.3% were found to be HIV-positive, up from 2.1% in 2004. The male-to-female ratio was 2.7:1. The largest percentage of co-infections were reported in people aged 25-34 years (47.8%). Recommended TB-HIV policies have been implemented in many of the countries. CONCLUSIONS: Case-finding has improved in many countries and the reported incidence of TB-HIV is increasing in the European region, particularly among young adults, although it remains low compared to other parts of the world. Strengthened coordination of existing TB and HIV services is still needed. PMID- 18410356 TI - Natural variation of gliding motility in a centimetre-scale population of Myxococcus xanthus. AB - A major challenge in microbial evolutionary ecology is to understand how fitness related traits vary in natural populations of microorganisms at defined spatial scales and subsequently to identify the forces that maintain such variation. The Gram-negative soil bacterium Myxococcus xanthus is a model system for the study of gliding motility, which is driven by two complementary motility systems in this species and is central to its social lifestyle. We tested whether the ecological context of a centimetre-scale M. xanthus population allows the coexistence of diverse motility-related phenotypes. Swarming rates among 26 clones isolated at the centimetre scale were found to vary greatly in multiple laboratory environments. This variation appears to be motility-specific, as it is not explained by a correlated variation in intrinsic growth rate. In contrast to the common reference strain DK1622, most isolates swarmed faster on hard agar than on soft agar, highlighting the difficulty of inferring species characteristics from laboratory reference strains. These isolates also varied greatly in swarm morphology and in the effect of nutrient limitation on swarming rate. Our results show that diverse swarming phenotypes can coexist in a small scale bacterial population. PMID- 18410354 TI - Delayed diagnosis of HIV infection and late initiation of antiretroviral therapy in the Swiss HIV Cohort Study. AB - OBJECTIVES: To investigate delayed HIV diagnosis and late initiation of antiretroviral therapy (ART) in the Swiss HIV Cohort Study. METHODS: Two sub populations were included: 1915 patients with HIV diagnosis from 1998 to 2007 and within 3 months of cohort registration (group A), and 1730 treatment-naive patients with CD4>or=200 cells/microL before their second cohort visit (group B). In group A, predictors for low initial CD4 cell counts were examined with a median regression. In group B, we studied predictors for CD4<200 cells/microL without ART despite cohort follow-up. RESULTS: Median initial CD4 cell count in group A was 331 cells/microL; 31% and 10% were <200 and <50 cells/microL, respectively. Risk factors for low CD4 count were age and non-White race. Homosexual transmission, intravenous drug use and living alone were protective. In group B, 30% initiated ART with CD4>or=200 cells/microL; 18% and 2% dropped to CD4 <200 and <50 cells/microL without ART, respectively. Sub-Saharan origin was associated with lower probability of CD4 <200 cells/microL without ART during follow-up. Median CD4 count at ART initiation was 207 and 253 cells/microL in groups A and B, respectively. CONCLUSIONS: CD4<200 cells/microL and, particularly, CD4<50 cells/microL before starting ART are predominantly caused by late presentation. Earlier HIV diagnosis is paramount. PMID- 18410357 TI - Cyanobacterial diversity in Salar de Huasco, a high altitude saline wetland in northern Chile: an example of geographical dispersion? AB - The diversity of Cyanobacteria in water and sediment samples from four representative sites of the Salar de Huasco was examined using denaturing gradient gel electrophoresis and analysis of clone libraries of 16S rRNA gene PCR products. Salar de Huasco is a high altitude (3800 m altitude) saline wetland located in the Chilean Altiplano. We analyzed samples from a tributary stream (H0) and three shallow lagoons (H1, H4, H6) that contrasted in their physicochemical conditions and associated biota. Seventy-eight phylotypes were identified in a total of 268 clonal sequences deriving from seven clone libraries of water and sediment samples. Oscillatoriales were frequently found in water samples from sites H0, H1 and H4 and in sediment samples from sites H1 and H4. Pleurocapsales were found only at site H0, while Chroococcales were recovered from sediment samples of sites H0 and H1, and from water samples of site H1. Nostocales were found in sediment samples from sites H1 and H4, and water samples from site H1 and were largely represented by sequences highly similar to Nodularia spumigena. We suggest that cyanobacterial communities from Salar de Huasco are unique - they include sequences related to others previously described from the Antarctic, along with others from diverse, but less extreme environments. PMID- 18410358 TI - Attention impairment in rolandic epilepsy: systematic review. AB - PURPOSE: Conflicting evidence about impairment of attention systems and the absence of a working model of attention has contributed to lack of clarity about comorbidity of attention problems in rolandic epilepsy (RE). Impairments in distributed attention systems may inform a disease model for RE, as well as direct interventions. METHODS: We used a systematic review of the relevant literature published in English from 1990 to 2006 to evaluate impairment in attention in RE. The Mirsky and Posner models of attention were evaluated for applicability, and studies were reviewed for design, instrumentation, and congruence with the Posner model of attention. RESULTS: Fourteen studies were identified: seven using a cross-sectional design (six active EEG abnormalities; one EEG remission) and seven longitudinal studies (abnormal EEGs and follow-up until normalized). According to the Posner model of attention, 12 studies employed measures that tapped the alerting network, 11 studies the orienting network, and eight the executive network. Nearly all controlled studies demonstrated impairments in all tested attention networks. In contrast, uncontrolled studies uniformly did not demonstrate impairments. Follow-up studies demonstrated complete or near complete resolution of attention impairments. DISCUSSION: The weight of evidence, defined as the majority of studies evaluated, suggests that all three attention systems are impaired in children with active centrotemporal spikes (CTS), implying a more widespread functional cortical disturbance in RE than previously held. These impairments resolve upon EEG remission, suggesting a common pathological basis to the autosomal dominant CTS trait. Sources of methodological variation are discussed with recommendations for future investigations. PMID- 18410359 TI - Epileptic encephalopathy with continuous spikes and waves during sleep in children with shunted hydrocephalus: a study of nine cases. AB - PURPOSE: We present a series of nine patients with early-onset hydrocephalus who had seizures and continuous spikes and waves during slow sleep (CSWS) associated with neurocognitive and motor deterioration. METHODS: Six boys and three girls aged 9-16 years (mean 11.3 years) were studied. [Correction added after online publication 12-Apr-2008: Number of girls and boys has been updated.] All patients underwent clinical examinations, electroencephalographic evaluations, neuroradiological imaging and neuropsychological assessment at first examination. Antiepileptic drugs (AEDs) were given in all cases and changed according to clinical and EEG evolution. RESULTS: Onset of epilepsy occurred from age 8 to 60 months (mean 19.6 months and median 14 months) with focal seizures with or without secondary generalized tonic-clonic seizures. Between ages 6 and 13 years (mean 10.4 years and median 8 years), hyperkinesia, aggressiveness, and poor socialization appeared in all nine cases. Reduced attention span, deterioration of language, and temporospatial disorientation were found in three of them. Negative myoclonus was found in two patients. The EEG showed CSWS. Response to change in treatment was good in all patients. None of the patients had relapses, seven of them have remained seizure free, and two continued having sporadic focal motor seizures during 2-5 years (mean 3 years) of follow-up. CONCLUSION: In children with early-onset hydrocephalus, particularly with behavioral and language disturbances and/or motor deterioration, CSWS should be considered. Periodic EEG recordings during sleep should be done in these children. The early identification of this particular electroclinical picture is crucial to start adequate treatment to avoid progressive cognitive deterioration. PMID- 18410360 TI - Temporal lobe epilepsy surgery and the quest for optimal extent of resection: a review. AB - The efficacy of surgery to treat drug-resistant temporal lobe epilepsy (TLE) has been demonstrated in a prospective randomized trial. It remains controversial which resection method gives best results for seizure freedom and neuropsychological function. This review of 53 studies addressing extent of resection in surgery for TLE identified seven prospective studies of which four were randomized. There is considerable variability between the intended resection and the volumetrically assessed end result. Even leaving hippocampus or amygdalum behind can result in seizure freedom rates around 50%. Most authors found seizure outcome in selective amygdalohippocampectomy (SAH) to be similar to that of lobectomy and there is considerable evidence for better neuropsychological outcome in SAH. Studies varied in the relationship between extent of mesial resection and seizure freedom, most authors finding no positive correlation to larger mesial resection. Electrophysiological tailoring saw no benefit from larger resection in 6 of 10 studies. It must be concluded that class I evidence concerning seizure outcome related to type and extent of resection of mesial temporal lobe structures is rare. Many studies are only retrospective and do not use MRI volumetry. SAH appears to have similar seizure outcome and a better cognitive outcome than TLR. It remains unclear whether a larger mesial resection extent leads to better seizure outcome. PMID- 18410361 TI - Clinical differences between patients with nonepileptic seizures who report antecedent sexual abuse and those who do not. AB - PURPOSE: To investigate clinical differences between patients with psychogenic nonepileptic seizures (PNES) who report antecedent sexual abuse, and patients who do not. METHODS: In a consecutive series of 176 patients with video-EEG confirmed PNES without epilepsy, we compared patients who reported antecedent sexual abuse with those who did not report sexual abuse, in respect of a range of demographic and clinical variables. RESULTS: Fifty-nine women (45%) and 5 men (11%) reported sexual abuse. Those reporting sexual abuse had earlier onset PNES (28.5 vs. 33.1 years, p = 0.0319) and greater delay from onset to diagnosis (median 5.2 vs. 3.2 years, p < 0.0137). They more often drew social security benefits (p = 0.0054) and were less often in cohabiting relationships (p = 0.0006). Those who reported sexual abuse had poorer mental health on a range of indicators. Their spells were more often "convulsive" (p = 0.0419), were more severe (p = 0.0011), were more likely to have emotional triggers (p = 0.0045) and to include prodromes (p = 0.0424) and flashbacks (p < 0.0001). A history of nocturnal spells (p = 0.0109), injury during spells (p = 0.0056), and incontinence during spells (p = 0.0083) were also more common in the patients reporting sexual abuse. DISCUSSION: Our results suggest that patients with PNES who report sexual abuse have more severe PNES, are more likely to have PNES with features that suggest epilepsy, and are psychiatrically more unwell than those who do not report sexual abuse. PMID- 18410362 TI - Language lateralization in temporal lobe epilepsy using functional MRI and probabilistic tractography. AB - PURPOSE: Language functional magnetic resonance imaging (fMRI) is used to noninvasively assess hemispheric language specialization as part of the presurgical work-up in temporal lobe epilepsy (TLE). White matter asymmetries on diffusion tensor imaging (DTI) may be related to language specialization as shown in controls and TLE. To refine our understanding of the effect of epilepsy on the structure-function relationships, we focused on the arcuate fasciculus (ArcF) and the inferior occipitofrontal fasciculus (IOF) and tested the relationship between DTI- and fMRI-based lateralization indices in TLE. METHODS: fMRI with three language tasks and DTI were obtained in 20 patients (12 right and 8 left TLE). The ArcF, a major language-related tract, and the IOF were segmented bilaterally using probabilistic tractography to obtain fractional anisotropy (FA) lateralization indices. These were correlated with fMRI-based lateralization indices computed in the inferior frontal gyrus (Pearson's correlation coefficient). RESULTS: fMRI indices were left-lateralized in 16 patients and bilateral or right-lateralized in four. In the ArcF, FA was higher on the left than on the right side, reaching significance in right but not in left TLE. We found a positive correlation between ArcF anisotropy and fMRI-based lateralization indices in right TLE (p < 0.009), but not in left TLE patients. No correlation was observed for the IOF. CONCLUSIONS: Right TLE patients with more left-lateralized functional activations also showed a leftward-lateralized arcuate fasciculus. The decoupling between the functional and structural indices of the ArcF underlines the complexity of the language network in left TLE patients. PMID- 18410363 TI - A case-control evaluation of the ketogenic diet versus ACTH for new-onset infantile spasms. AB - PURPOSE: ACTH is currently the standard first-line therapy for new-onset infantile spasms, but it has significant side effects. We hypothesized the ketogenic diet (KD), previously reported as beneficial for intractable infantile spasms, would have similar efficacy, but better tolerability than ACTH when used first-line. METHODS: We conducted a retrospective chart review of all infants started on the KD (n = 13) and high-dose ACTH (n = 20) for new-onset infantile spasms at our institution since 1996. RESULTS: Infants were spasm-free in 8 of 13 (62%) infants treated with the KD within 1 month, compared to 18 of 20 (90%) treated initially with ACTH, p = 0.06. When effective, median time to spasm freedom was similar between ACTH and the KD (4.0 vs. 6.5 days, p = 0.18). Those treated with ACTH were more likely to have a normal EEG at 1 month (53% vs. 9%, p = 0.02), however, use of the KD led to EEG normalization within 2-5 months in all eight who became spasm-free. In the five children in whom the KD was unsuccessful, four became spasm-free subsequently with ACTH or topiramate immediately. Side effects (31% vs. 80%, p = 0.006) and relapse rate after initial success (12.5% vs. 33%, p = 0.23) were lower with the KD. DISCUSSION: In this retrospective study, the KD stopped spasms in nearly two-thirds of cases, and had fewer side effects and relapses than ACTH. ACTH normalized the EEG more rapidly, however. Further prospective study of the KD as, with a 2-week time limit if unsuccesful, first-line therapy for infantile spasms is warranted. PMID- 18410364 TI - Neural basis of photo/chromatic sensitivity in adolescence. AB - PURPOSE: To determine a psychophysiological basis for age visual sensitivity to chromatic and achromatic stimuli. METHODS: We investigated the effects of achromatic and four isoluminant color combinations (blue/red, blue/green, green/red, and blue/yellow), luminance ratio changes in color combinations (blue/red; 1:1, 3:4, 4:3) and contrast changes (3 to 100%) on steady-state electroretinograms (ERGs) and visual evoked potentials (VEPs) in 32 healthy teenagers and 30 young adults. RESULTS: We found that (1) dual peaks at 9 and 18 Hz with a dip at 12 Hz were observed in VEPs with all isoluminant color combinations, (2) VEP responses were significantly enhanced and the 12-Hz dip became unclear with luminance ratio changes between two colors with a nonantagonistic relationship (blue/red), and (3) VEP amplitudes were significantly increased when the contrast was increased. These characteristics were more evident in teenagers than young adults; however, ERGs were qualitatively similar between the two groups. DISCUSSION: The visual cortex is differently modulated by different color-luminance combinations, and higher sensitivity to color-luminance combinations in the visual cortex in teenagers is responsible for the high prevalence of photo/chromatic sensitivity in adolescence. PMID- 18410365 TI - Decreased hippocampal volume on MRI is associated with increased extracellular glutamate in epilepsy patients. AB - PURPOSE: Temporal lobe epilepsy (TLE) is associated with smaller hippocampal volume and with elevated extracellular (EC) glutamate levels. We investigated the relationship between the hippocampal volume and glutamate in refractory TLE patients. METHODS: We used quantitative MRI volumetrics to measure the hippocampal volume and zero-flow microdialysis to measure the interictal glutamate, glutamine, and GABA levels in the epileptogenic hippocampus of 17 patients with medication-resistant epilepsy undergoing intracranial EEG evaluation. The relationships between hippocampal volume, neurochemical levels, and relevant clinical factors were examined. RESULTS: Increased EC glutamate in the epileptogenic hippocampus was significantly related to smaller ipsilateral (R(2)= 0.75, p < 0.0001), but not contralateral hippocampal volume when controlled for glutamine and GABA levels, and for clinical factors known to influence hippocampal volume. Glutamate in the atrophic hippocampus was significantly higher (p = 0.008, n = 9), with the threshold for hippocampal atrophy estimated as 5 microM. GABA and glutamine levels in the atrophic and nonatrophic hippocampus were comparable. Decreased hippocampal volume was related to higher seizure frequency (p = 0.008), but not to disease duration or febrile seizure history. None of these clinical factors were related to the neurochemical levels. CONCLUSIONS: We provide evidence for a significant association between increased EC glutamate and decreased ipsilateral epileptogenic hippocampal volume in TLE. Future work will be needed to determine whether the increase in glutamate has a causal relationship with hippocampal atrophy, or whether another, yet unknown factor results in both. This work has implications for the understanding and treatment of epilepsy as well as other neurodegenerative disorders associated with hippocampal atrophy. PMID- 18410366 TI - The FLEP scale in diagnosing nocturnal frontal lobe epilepsy, NREM and REM parasomnias: data from a tertiary sleep and epilepsy unit. AB - PURPOSE: To test the usefulness of the FLEP scale in diagnosing nocturnal frontal lobe epilepsy (NFLE), arousal parasomnias, and REM sleep behavior disorder (RBD). METHODS: The FLEP scale was applied to 71 subjects (60 male; 11 female; aged 54 +/- 21) referred to an outpatient's sleep and epilepsy unit for diagnostic assessment of nocturnal motor-behavioral episodes, which turned to be arousal parasomnias (11 subjects), NFLE (14 subjects), or idiopathic RBD (46 subjects), based on the findings of in-lab full night video polysomnography with extended EEG montages. RESULTS: The sensitivity of the scale as a diagnostic test for NFLE was 71.4%, the specificity 100%, the positive predictive value 100%, and the negative predictive value 91.1%. The FLEP scale gave an incorrect diagnosis in 4/71 (5.6%) of the cases, namely NFLE patients with episodes of nocturnal wandering, and uncertain diagnostic indications in 22/71 subjects (30.9%). CONCLUSIONS: The FLEP scale shows high positive and negative predictive values in diagnosing NFLE versus arousal parasomnias and RBD. However, the scale is associated with a real risk of misdiagnosis in some patients and gives uncertain indications in about one-third of cases, mainly RBD. Our investigation highlights the inadequacy of some of the items in the scale. The item investigating wandering, as presently formulated, may be unable to distinguish nocturnal wandering from sleepwalking. The items about "recall" and "clustering" of the events throughout the night may increase the likelihood of mistaking RBD for seizures. Further testing of the reliability of the FLEP scale items appears to be needed. PMID- 18410367 TI - Seizure remission and relapse in adults with intractable epilepsy: a cohort study. AB - PURPOSE: To investigate the cumulative probabilities of >or=12 month seizure remission and seizure relapse following remission, and to test the associations of clinical characteristics with these two study end points in a prevalence cohort of intractable adult epilepsy patients during medical management. METHODS: A retrospective cohort study of intractable epilepsy patients seen in 2001 at a single center was conducted. Kaplan-Meier analysis was used to estimate the cumulative probabilities of seizure remission and subsequent seizure relapse. Cox proportional hazards models were used to estimate the association (1) between clinical factors and >or=12 month seizure remission and (2) between clinical factors and seizure relapse following remission. RESULTS: One hundred eighty seven subjects met the eligibility criteria for intractable epilepsy. The estimate of probability of remission was about 4% per year. Seizure remission was temporary for some individuals, as 5 out of 20 subjects with remission ultimately relapsed. No clinical factors predicted the likelihood of achieving >or=12 month seizure remission or subsequent seizure relapse. DISCUSSION: Some people with intractable epilepsy achieve >or=12 month seizure remission during medical treatment. Remission, however, is only temporary for some individuals. We were unable to identify clear predictors for remission. PMID- 18410368 TI - An unusual pathologic feature and phenotype associated with familial hyperkalemic periodic paralysis. PMID- 18410369 TI - Assessment of idiopathic normal pressure patients in neurological practice: the role of lumbar infusion testing for referral of patients to neurosurgery. AB - BACKGROUND AND PURPOSE: In neurological practice patients with tentative idiopathic normal pressure hydrocephalus (iNPH) usually are referred to neurosurgery based on clinical and radiological findings. Hydrodynamic assessment using lumbar infusion testing might be helpful in selecting patients. To retrospectively analyse lumbar infusion tests done in neurological practice in iNPH patients to see how infusion test results relate to the clinical course and shunt response. MATERIALS AND METHODS: Sixty-three consecutive patients with Possible/Probable iNPH were tested during a 1-year period. The pre-operative lumbar infusion tests were assessed according to two strategies: (i) Determining the resistance to cerebrospinal fluid (CSF) outflow (R(out)). (ii) Quantification of the CSF pressure (CSFP) pulsatility during lumbar infusion (Q(pulse)). The results were related to the prospectively followed clinical course and shunt response after 12 months. RESULTS: The lumbar infusion-derived parameters R(out) and Q(pulse) related weakly. Shunt response after 12 months was not related to R(out), but was highly related to the Q(pulse). False negative results of lumbar infusion testing were observed in 16% of the patients. DISCUSSION: In neurological practice lumbar infusion testing may be useful for determining which patients to refer to neurosurgery. Our data favour determination of CSFP pulsatility (Q(pulse)) rather than R(out) for prediction of shunt response. PMID- 18410370 TI - A descriptive study on constructional impairment in frontotemporal dementia and Alzheimer's disease. AB - BACKGROUND AND PURPOSE: Frontotemporal dementia (FTD) is a degenerative disorder characterized, in its frontal variant, by psychiatric onset, deficits in executive functions and sparing of memory and visuo-spatial abilities. Studies on visuo-spatial abilities in FTD and Alzheimer's disease (AD) have mainly focused on constructional abilities (CA) and have yielded contrasting data, often depending either on the task used or biases in the clinical selection of patients. The aim of our study was to evaluate the frequency of the CA impairment in FTD-fv and AD patients at the early stage of the disease, and to verify if some distinctive features of this impairment can characterize each pathology. METHODS: The copy-drawing task (Rey Complex Figure, RCF) of 41 patients with probable mild AD and 15 patients with probable mild FTD-fv were analysed by means of quantitative and qualitative indices. RESULTS: Data showed that both AD and FTD patients displayed similar RCF scores, as well as execution strategies, type of errors and global analysis. The FTD group only had a significant deficit in the analysis of inner details. PMID- 18410372 TI - Monitoring treatment with cyclosporine microemulsion in myasthenia gravis. AB - PURPOSE: To examine whether the monitoring of cyclosporine (CsA) blood concentrations is of benefit in CsA microemulsion pre-concentrate (MEPC) therapy for myasthenia gravis (MG). METHODS: We measured CsA blood concentrations both 2 h after administration (C2) and immediately before administration (C0) and examined associations with changes to clinical parameters in 20 MG patients treated with CsA MEPC in an unblinded, 6-month prospective open trial. RESULTS: Initial dose of CsA MEPC (4.7 +/- 0.5 mg/kg/day) provided both high C2 levels and safe C0 levels. Disease severity, daily dose of prednisolone, acetylcholine receptor-antibody titre levels and levels of interleukin-2 production by peripheral blood mononuclear cells were significantly reduced following treatment with CsA MEPC. A significant correlation existed between C2 levels following the initial dose and clinical improvement in responder MG patients. C0 levels were significantly higher in patients who exhibited increased serum creatinine or hypertension compared with patients free from side effects. Body mass index of individual patients was significantly correlated with C0 level, and may thus offer a useful marker to predict C0 levels. DISCUSSION: CsA MEPC was effective at suppressing symptoms and T-cell-dependent pathogenesis of MG, and monitoring of C2 and C0 levels can be useful to estimate efficacy and safety of the drug. PMID- 18410371 TI - A 71-nucleotide deletion in the periaxin gene in a Romani patient with early onset slowly progressive demyelinating CMT. AB - BACKGROUND: Mutations in the periaxin (PRX) gene cause autosomal recessive demyelinating neuropathy Charcot-Marie-Tooth (CMT) type 4F. To date, 10 non-sense or frameshift PRX mutations have been reported in patients with early-onset neuropathy and further disease course consistent with either Dejerine-Sottas neuropathy or slow-progressive demyelinating CMT. METHODS: We sequenced 59 patients from 55 Czech families including four unrelated patients of Romani (Gypsy) origin with early-onset CMT displaying decreased nerve conduction velocities. RESULTS: We identified a novel homozygous mutation c.3286_3356del71 (K1095fsX18) in one Romani patient showing very slow disease progression. Amongst non-Romani Czech CMT patients, PRX mutations have been proven to be very rare. PMID- 18410373 TI - Brain barrier dysfunction in Cuban epidemic optic neuropathy. AB - BACKGROUND AND PURPOSE: There are practically no references to cerebrospinal fluid (CSF) studies in tropical or nutritional neuropathies. In the present paper we present the results of CSF studies in patients with Cuban Epidemic Optic Neuropathy (CEON) during epidemic and endemic periods, with an appraisal as to the contribution of brain barriers' function in the pathophysiology of this disease. METHODS: Two hundred and five patients with CEON were studied during the epidemic period (1992-1993) and 12 patients outside the outbreak (1995-1997). CSF protein determination and electrophoresis were carried out, as well as serum and CSF albumin and immunoglobulin G (IgG) quantitation for calculating IgG and Q(alb) indexes, in order to evaluate intrathecal IgG synthesis and the permeability of the blood-CSF barrier (B-CSF B). RESULTS: One fourth of the patients had increased permeability of the B-CSF B, but damage was more frequent between 16 and 60 days from onset of disease, disappearing after 120 days. B-CSF B dysfunction was more prevalent in patients with severe neurological impairment, although it was not related to the severity of ophthalmological damage. The group of patients studied outside of the outbreak (endemic period) showed similar results. DISCUSSION: The possible association of increased permeability of the B CSF B with oxidative stress, which lies on the basis of this epidemic outbreak, is discussed. PMID- 18410374 TI - Hypokalemic thyrotoxic periodic paralysis: clinical characteristics and predictors of recurrent paralytic attacks. AB - BACKGROUND AND PURPOSE: To study the clinical characteristics of hypokalemic thyrotoxic periodic paralysis (hoTPP) and identify the predictors of recurrent paralytic attacks before achieving the euthyroid status. METHODS: We retrospectively analyzed 45 hoTPP patients who were admitted during the 7-year study period. RESULTS: A tendency towards male predominance was observed among the 45 patients (91.1%, 41/45). The mean onset age was 32.9 +/- 10.0 years (range: 16-54 years). No significant differences were observed in the onset age between male and female patients. Precipitating factors included rest/sleep at night, hot weather, upper respiratory tract infections (URIs), and excessive physical activities. Atypical weakness was observed in nine (20%, 9/45) patients. One patient initially diagnosed with sporadic periodic paralysis eventually developed hoTPP. DISCUSSION: In provocative tests, hypokalemia was not a consistent finding during paralytic attacks. Before achieving the euthyroid status, the rate of recurrent attacks was as high as 62.2%, and peaked in the first 3 months after hoTPP was diagnosed. Patients with URIs exhibited a higher incidence of recurrent paralytic attacks than those without (odds ratio = 13.00; 95% confidence interval = 1.08-156.08; P = 0.04). PMID- 18410375 TI - Correlation between sleep and cognitive functions after hemispheric ischaemic stroke. AB - BACKGROUND: The aim of this study was to test the hypothesis of a link between sleep and cognitive functions, particularly memory and attention, after stroke. METHODS: We studied 11 consecutive patients with first-ever hemispheric ischaemic stroke within eight days after symptoms onset and nine of them at least three months after stroke. Sleep EEG was recorded with a portable system. Cognitive functions were assessed using a standardized battery of tests allowing the estimation of the most relevant domains of cognition. Five age-matched healthy subjects served as controls. RESULTS: The patients were aged 43 +/- 12 years (18 59). In five patients stroke was right-sided and in six patients left-sided. In the acute stroke phase a correlation between attention and amounts of slow wave sleep (SWS), Rapid eye movement (REM) sleep and sleep efficiency was found. In the recovery phase verbal/figural memory and attention significantly improved in most patients. Furthermore, an association between (i) verbal/figural (non verbal) memory and amounts of SWS, REM sleep and sleep efficiency, and between (ii) attention and sleep efficiency was observed. CONCLUSIONS: The results point to a link between sleep and cognitive functions and their recovery after hemispheric stroke. Further studies are needed to determine the specific nature of this link. PMID- 18410376 TI - Event-related delta oscillatory responses of Alzheimer patients. AB - BACKGROUND AND PURPOSE: Alzheimer type of dementia (AD) is the most common neuropsychiatric morbidity in elderly individuals. Event-related oscillations (ERO) provide an useful tool for detecting subtle abnormalities of cognitive processes with high temporal resolution. METHODS: In the present report, event related oscillations of patients with AD were analyzed by using a visual oddball paradigm. A total of 22 mild probable AD subjects according to NINCDS-ADRDA criteria and 20 age-, gender-, and education-matched healthy control subjects were compared. AD group consisted from 11 untreated patients and 11 patients treated with cholinesterase inhibitor. Oscillatory responses were recorded from 13 scalp electrodes. RESULTS: Significant differences in delta frequency range were seen between the groups by using repeated measures of anova analysis [F(9.120) = 2.228; P = 0.022]. Post-hoc analyses using Wilcoxon test showed that at mid- and left central regions, (Cz, C3) peak amplitudes of delta responses of healthy subjects were significantly higher than either group. Also cholinesterase inhibitors did not have effect on delta oscillatory responses. CONCLUSIONS: Our findings imply that the delta oscillatory responses at central locations are highly instable in mild probable AD patients regardless of treatment when compared to the healthy aged controls. This study supports the importance of oscillatory event-related potentials for investigating AD brain dynamics. PMID- 18410377 TI - Invading populations of an ornamental shrub show rapid life history evolution despite genetic bottlenecks. AB - Human-mediated species introductions offer opportunities to investigate when and how non-native species to adapt to novel environments, and whether evolution has the potential to contribute to colonization success. Many long-established introductions harbour high genetic diversity, raising the possibility that multiple introductions of genetic material catalyze adaptation and/or the evolution of invasiveness. Studies of nascent invasions are rare but crucial for understanding whether genetic diversity facilitates population expansion. We explore variation and evolution in founder populations of the invasive shrub Hypericum canariense. We find that these introductions have experienced large reductions in genetic diversity, but that increased growth and a latitudinal cline in flowering phenology have nevertheless evolved. These life history changes are consistent with predictions for invasive plants. Our results highlight the potential for even genetically depauperate founding populations to adapt and evolve invasive patters of spread. PMID- 18410378 TI - Bioimaging of the unbalanced expression of microRNA9 and microRNA9* during the neuronal differentiation of P19 cells. AB - Generally, the 3'-end of the duplex microRNA (miR) precursor (pre-miR) is known to be stable in vivo and serve as a mature form of miR. However, both the 3'-end (miR9) and 5'-end (miR9*) of a brain-specific miR9 have been shown to function biologically in brain development. In this study, real-time PCR analysis and in vitro/in vivo bioluminescent imaging demonstrated that the upstream region of a primary miR9-1 (pri-miR9-1) can be used to monitor the highly expressed pattern of endogenous pri-miR9-1 during neurogenesis, and that the Luciferase reporter gene can image the unequal expression patterns of miR9 and miR9* seen during the neuronal differentiation of P19 cells. This demonstrates that our bioimaging system can be used to study the participation of miRs in the regulation of neuronal differentiation. PMID- 18410380 TI - Structural bases for recognition of Anp32/LANP proteins. AB - The leucine-rich repeat acidic nuclear protein (Anp32a/LANP) belongs to a family of evolutionarily-conserved phosphoproteins involved in a complex network of protein-protein interactions. In an effort to understand the cellular role, we have investigated the mode of interaction of Anp32a with its partners. As a prerequisite, we solved the structure in solution of the evolutionarily conserved N-terminal leucine-rich repeat (LRR) domain and modeled its interactions with other proteins, taking PP2A as a paradigmatic example. The interaction between the Anp32a LRR domain and the AXH domain of ataxin-1 was probed experimentally. The two isolated and unmodified domains bind with very weak (millimolar) affinity, thus suggesting the necessity either for an additional partner (e.g. other regions of either or both proteins or a third molecule) or for a post translational modification. Finally, we identified by two-hybrid screening a new partner of the LRR domain, i.e. the microtubule plus-end tracking protein Clip 170/Restin, known to regulate the dynamic properties of microtubules and to be associated with severe human pathologies. PMID- 18410379 TI - Pathways and products for the metabolism of vitamin D3 by cytochrome P450scc. AB - Cytochrome P450scc (CYP11A1) can hydroxylate vitamin D3 to produce 20 hydroxyvitamin D3 and other poorly characterized hydroxylated products. The present study aimed to identify all the products of vitamin D3 metabolism by P450scc, as well as the pathways leading to their formation. Besides 20 hydroxyvitamin D3, other major metabolites of vitamin D3 were a dihydroxyvitamin D3 and a trihydroxyvitamin D3 product. The dihydroxyvitamin D3 was clearly identified as 20,23-dihydroxyvitamin D3 by NMR, in contrast to previous reports that postulated hydroxyl groups in positions 20 and 22. NMR of the trihydroxy product identified it as 17alpha,20,23-trihydroxyvitamin D3. This product could be directly produced by P450scc acting on 20,23-dihydroxyvitamin D3, confirming that hydroxyl groups are present at positions 20 and 23. Three minor products of D3 metabolism by P450scc were identified by MS and by examining their subsequent metabolism by P450scc. These products were 23-hydroxyvitamin D3, 17alpha hydroxyvitamin D3 and 17alpha,20-dihydroxyvitamin D3 and arise from the three P450scc-catalysed hydroxylations occurring in a different order. We conclude that the major pathway of vitamin D3 metabolism by P450scc is: vitamin D3 --> 20 hydroxyvitamin D3 --> 20,23-dihydroxyvitamin D3 --> 17alpha,20,23 trihydroxyvitamin D3. The major products dissociate from the P450scc active site and accumulate at a concentration well above the P450scc concentration. Our new identification of the major dihydroxyvitamin D3 product as 20,23-dihydroxyvitamin D3, rather than 20,22-dihydroxyvitamin D3, explains why there is no cleavage of the vitamin D3 side chain, unlike the metabolism of cholesterol by P450scc. PMID- 18410381 TI - Identification of a novel matrix protein contained in a protein aggregate associated with collagen in fish otoliths. AB - In the biomineralization processes, proteins are thought to control the polymorphism and morphology of the crystals by forming complexes of structural and mineral-associated proteins. To identify such proteins, we have searched for proteins that may form high-molecular-weight (HMW) aggregates in the matrix of fish otoliths that have aragonite and vaterite as their crystal polymorphs. By screening a cDNA library of the trout inner ear using an antiserum raised against whole otolith matrix, a novel protein, named otolith matrix macromolecule-64 (OMM 64), was identified. The protein was found to have a molecular mass of 64 kDa, and to contain two tandem repeats and a Glu-rich region. The structure of the protein and that of its DNA are similar to those of starmaker, a protein involved in the polymorphism control in the zebrafish otoliths [Sollner C, Burghammer M, Busch-Nentwich E, Berger J, Schwarz H, Riekel C & Nicolson T (2003) Science302, 282-286]. (45)Ca overlay analysis revealed that the Glu-rich region has calcium binding activity. Combined analysis by western blotting and deglycosylation suggested that OMM-64 is present in an HMW aggregate with heparan sulfate chains. Histological observations revealed that OMM-64 is expressed specifically in otolith matrix-producing cells and deposited onto the otolith. Moreover, the HMW aggregate binds to the inner ear-specific short-chain collagen otolin-1, and the resulting complex forms ring-like structures in the otolith matrix. Overall, OMM 64, by forming a calcium-binding aggregate that binds to otolin-1 and forming matrix protein architectures, may be involved in the control of crystal morphology during otolith biomineralization. PMID- 18410382 TI - Collective behavior in gene regulation: the cell is an oscillator, the cell cycle a developmental process. AB - The finding of a genome-wide oscillation in transcription that gates cells into S phase and coordinates mitochondrial and metabolic functions has altered our understanding of how the cell cycle is timed and how stable cellular phenotypes are maintained. Here we present the evidence and arguments in support of the idea that everything oscillates, and the rationale for viewing the cell as an attractor from which deterministic noise can be tuned by appropriate coupling among the many feedback loops, or regulons, that make up the transcriptional respiratory attractor cycle. The existence of this attractor also explains many of the dynamic macroscopic properties of the cell cycle and appears to be the timekeeping oscillator in both cell cycles and circadian rhythms. The path taken by this primordial oscillator in the course of differentiation or drug response may involve period-doubling behavior. Evidence for a relatively high-frequency timekeeping oscillator in yeast and mammalian cells comes from expression array analysis, and GC/MS in the case of yeast, and primarily from macroscopic measures of phase response to perturbation in the case of mammalian cells. Low-amplitude, genome-wide oscillations, a ubiquitous but often unrecognized attribute of phenotype, may be a source of seemingly intractable biological noise in microarray and proteomic studies. These oscillations in transcript and protein levels and the repeated cycles of synthesis and degradation they require, represent a high energy cost to the cell which must, from an evolutionary point of view, be recovered as essential information. We suggest that the information contained in this genome-wide oscillation is the dynamic code that organizes a stable phenotype from an otherwise passive genome. PMID- 18410383 TI - Collective behavior in gene regulation: post-transcriptional regulation and the temporal compartmentalization of cellular cycles. AB - Self-sustained oscillations are perhaps the most studied objects in science. The accomplishment of such a task reliably and accurately requires the presence of specific control mechanisms to face the presence of variable and largely unpredictable environmental stimuli and noise. Self-sustained oscillations of transcript abundance are, in fact, widespread and are not limited to the reproductive cycle but are also observed during circadian rhythms, metabolic cycles, developmental cycles and so on. To date, much of the literature has focused on the transcriptional machinery underlying control of the basic timing of transcript abundance. However, mRNA abundance is known to be regulated at the post-transcriptional level also and the relative contribution of the two mechanisms to gene-expression programmes is currently a major challenge in molecular biology. Here, we review recent results showing the relevance of the post-transcriptional regulation layer and present a statistical reanalysis of the yeast metabolic cycle using publicly available gene-expression and RNA-binding data. Taken together, the recent theoretical and experimental developments reviewed and the results of our reanalysis strongly indicate that regulation of mRNA stability is a widespread, phase-specific and finely tuned mechanism for the multi-layer control of gene expression needed to achieve high flexibility and adaptability to external and internal signals. PMID- 18410384 TI - Collective behavior in gene regulation: metabolic clocks and cross-talking. AB - Biological functions governed by the circadian clock are the evident result of the entrainment operated by the earth's day and night cycle on living organisms. However, the circadian clock is not unique, and cells and organisms possess many other cyclic activities. These activities are difficult to observe if carried out by single cells and the cells are not coordinated but, if they can be detected, cell-to-cell cross-talk and synchronization among cells must exist. Some of these cycles are metabolic and cell synchronization is due to small molecules acting as metabolic messengers. We propose a short survey of cellular cycles, paying special attention to metabolic cycles and cellular cross-talking, particularly when the synchronization of metabolism or, more generally, cellular functions are concerned. Questions arising from the observation of phenomena based on cell communication and from basic cellular cycles are also proposed. PMID- 18410385 TI - Cell dynamics and the onset of collective behaviour in gene regulation. PMID- 18410386 TI - Evaluating Emdogain and healing of replanted teeth using an intra-individual experimental-control study design. AB - The purpose of the present investigation was to use an intra-individual experimental-control study design to explore if application of Emdogain prior to re-plantation after a dry extra-alveolar period of 60 min would promote a favorable healing of the periodontal ligament cells. Ten patients, for whom already decisions had been taken to extract two maxillary premolars because of crowding, participated in the study. The teeth were extracted and endodontic treatment was performed extra orally. The experimental tooth and its alveolar socket were covered with Emdogain prior to replantation. The contra lateral tooth served as a control and was replanted without any prior treatment. The teeth were stabilized with a retainer for 3-7 days and the patients were followed up every third week. After 13 weeks, the teeth were finally extracted and prepared for histological examination. Radiographs were taken before the study period, at day 29 and prior to the final extraction. The results were in favor of Emdogain, but the overall difference between the Emdogain-treated tooth and its control was rather small, and it seemed questionable if the registered differences could be of any obvious practical clinical importance. Histologically, all the teeth showed some degree of pathology after such a long dry extra-oral time and the outcome seemed to be more correlated to the individual than to the treatment. Given more favorable conditions regarding storage medium and/or extra-oral time, Emdogain might still be of value for an uncomplicated healing after replantation. PMID- 18410387 TI - In vitro evaluation of casein phosphopeptide-amorphous calcium phosphate as a potential tooth transport medium: viability and apoptosis in L929 fibroblasts. AB - Casein phosphopeptides (CPP) are derived from casein, which accounts for 80% of the total protein in bovine milk . The purpose of this in vitro study was to evaluate the potential use of a CPP-amorphous calcium phosphate (CPP-ACP) preparation as a transport medium for avulsed teeth. L929 fibroblastic cell line was plated in 24-well culture plates. Following incubation, the cells were treated with 10(-3), 10(-4), 10(-6), 10(-8), 10(-12) dilutions of a water-based CPP-ACP paste (Tooth Mousse, GC Corp., Tokyo, Japan). Untreated cells served as controls. The L929 cells were counted at the 1st, 3rd and 7th days. Propidium iodide/acridine orange staining was used to assess apoptosis of treated cells and of the positive control. For each concentration (dilution), statistical analysis of cell survival within time was performed using two-way analysis of variance (ANOVA, P = 0.05). One way ANOVA and Tukey tests were applied to compare the effect of different concentrations on cell survival at each evaluation day (P = 0.05). Except for the 10(-3) and 10(-4) dilutions, all groups demonstrated an increase in cell numbers at days 1 and 3, followed by a decrease at day 7. Irrespective of the increase or decrease in cell viability, time-dependent changes for each dilution group were significantly different. Cells in the 10(-3) and 10(-4) dilution groups demonstrated a rapid apoptotic response. A relatively few number of apoptotic cells were observed in the 10(-6) and 10(-8) dilution groups, while no sign of apoptosis was evident in the 10(-12) dilution group and control. These results suggest that when highly diluted, the tested CPP-ACP preparation may help preserve L929 cell viability in the short term without inducing apoptosis. PMID- 18410388 TI - Treatment of root surface in delayed tooth replantation: a review of literature. AB - The time elapsed between a trauma and tooth replantation usually ranges from 1 to 4 h. The chances of root surface damage are higher when tooth replantation is not performed immediately or if the avulsed tooth is not stored in an adequate medium. This invariably leads to necrosis of pulp tissue, periodontal ligament cells and cementum, thus increasing the possibility of root resorption, which is the main cause of loss of replanted teeth. This paper presents a comprehensive review of literature on root surface treatments performed in cases of delayed tooth replantation with necrotic cemental periodontal ligament. Journal articles retrieved from PubMed/MedLine, Bireme and Scielo databases were reviewed. It was observed that, when there are no periodontal ligament remnants and contamination is under control, replacement resorption and ankylosis are the best results and that, although these events will end up leading to tooth loss, this will happen slowly with no loss of the alveolar ridge height, which is important for future prosthesis planning. PMID- 18410389 TI - Etiology and environment of dental injuries in 12- to 14-year-old Ontario schoolchildren. AB - This study reports on the etiology and environment where dental injuries occurred and assesses the relationship between dental trauma, socio-economic status and dental caries experience. A population-based, matched case-comparison study was undertaken in 30 schools in two Ontario communities. Dental hygienists calibrated in the use of the Dental Trauma Index (DTI) screened 2422 children aged 12 and 14 years using DTI and Decayed, Missing and Filled Teeth indices. Cases (n = 135) were children with evidence of dental injury. Controls (n = 135) were children randomly selected after screening and matched with cases according to age and gender. Questionnaires were mailed to parents and children. Prevalence of dental injury was 11.4%, mostly minor injuries 63.7% (enamel fracture not involving dentin), affecting one upper central incisor (70.4%). The mean age at the time of dental injuries was 9.5 years (SD = 1.49; range: 6-13 years). Dental trauma most often occurred among boys at school because of falls or while playing sports. The relationship between dental injuries and the socio-economic indicators chosen was not statistically significant. However, a statistically significant direct relationship (P < 0.001) was shown between increased caries experience and dental injuries. This should focus attention on possible common-risk factors such as health-related behavioral problems that may affect both dental disease and dental injuries. PMID- 18410390 TI - Impact of treated and untreated dental injuries on the quality of life of Ontario school children. AB - A population-based, matched case-comparison study was undertaken in 30 schools in two Ontario communities to measure the impact of dental trauma on quality of life (QoL) in Canadian school children. Dental hygienists screened 2422 children aged 12-14 years using the dental trauma index, the decayed, missing and filled teeth index (DMFT) and the aesthetic component of the index of orthodontic treatment needs (AC-IOTN). Cases (n = 135) were children with evidence of previous dental trauma. Controls (n = 135) were classmates matched for age and gender. Oral health-related QoL was assessed using mailed Child Perception Questionnaires (CPQ(11-14)) completed by all children. Data were analyzed using simple and multiple conditional logistic regressions after adjusting for DMFT and AC-IOTN, CPQ(11-14), overall impact and item-specific impacts. Approximately 64% of injuries were untreated enamel fractures and just over 30% were previously injured restored teeth. Untreated children experienced more chewing difficulties (P = 0.026), avoided smiling (P = 0.029) and experienced affected social interactions (P = 0.032) compared with their non-injured peers. When treated and non-injured groups were compared, the only statistically significant effect was difficulty in chewing (P = 0.038). Injured children who were untreated experienced more social impact than their non-injured peers. Restoration of injured teeth improved aesthetics and social interactions but functional deficiencies persisted as a result of periodontal or pulpal pain. PMID- 18410391 TI - Elementary school staff knowledge about dental injuries. AB - Elementary school staff can play a crucial role in managing traumatic dental injuries (TDIs) because they are often in proximity to children and are frequently called upon to assist with children's accidents. International studies reveal that elementary school personnel have little knowledge about emergency dental care and management. The purpose of this study was to assess the knowledge, practice and experience regarding TDIs among a sample of elementary school personnel in the USA. Assessment was performed using a demographic questionnaire and a newly developed TDI survey instrument. Results revealed a wide distribution of responses. Overall, dental trauma knowledge among this group was poor. The majority of respondents were not well-versed regarding TDIs, their management, the benefits of timely care or treatment costs. However, staff reported a keen interest in receiving more TDI information and training. TDI education and management are needed among all elementary school staff members to improve the prognosis of these accidents when they occur. PMID- 18410392 TI - Fracture resistance and histological findings of immature teeth treated with mineral trioxide aggregate. AB - The objective of the present study was to test the hypothesis that the fracture strength of calcium hydroxide and mineral trioxide aggregate (MTA)-filled immature teeth decreased over time. Immature mandibular incisors from sheep were extracted and the pulps were extirpated using an apical approach with a barbed broach, and the teeth were divided into three experimental groups. Group 1: untreated teeth. Group 2: the root canals were filled with calcium hydroxide paste. Group 3: the root canals were filled with MTA. All specimens were kept in saline with 1% antibiotics at 4 degrees C for certain periods of time: 2 weeks, 2 months, and 1 year. Then they were tested for fracture strength in an Instron testing machine. The results were subjected to statistical analysis by the Tukey Kramer tests. A P-value (<0.05) was considered statistically significant. One tooth from each group was selected randomly for a histological study, examining matrix metalloproteinases (MMP2 and MMP14) and tissue inhibitor of metalloproteinase (TIMP). The results showed the mean fracture strengths decreased over time for all the three groups. Although the untreated teeth showed the highest value (45.5 MPa) at 2 weeks, the fracture strengths decreased significantly after 2 months (P < 0.05). On the other hand, the teeth treated with calcium hydroxide or MTA decreased, but not significantly over time (P > 0.05). For the MTA-treated teeth, the fracture strengths were not found significantly different from the untreated or calcium hydroxide-treated teeth at 2 weeks or 2 months (P > 0.05). However, the strength was significantly higher in the MTA group compared with the other two groups after 1 year (P < 0.05). Immunofluorescence images revealed expression of collagen type 1, MMP-2 and MMP 14 in both untreated and endodontically treated teeth. However, TIMP-2 was only observed in the MTA-treated teeth. In conclusion, the teeth with root treatment with MTA showed the highest fracture resistance at 1 year (P < 0.05). An explanation could be that MTA induced the expression of TIMP-2 in the dentin matrix and thereby possibly prevented destruction of the collagen matrix. PMID- 18410393 TI - Anal intraepithelial neoplasia in HIV infection. AB - Human papillomavirus (HPV) infections belong to the most common sexually transmitted infections worldwide. While the immune system eliminates most HPV infections over time in immunocompetent individuals, HPV infections tend to persist in immunodeficient individuals. In HIV-infected men who have sex with men (MSM), anal HPV prevalence is more than 90% and infections with multiple HPV types are common. Consequently, HPV-associated anogenital malignancies occur with high frequency in patients with HIV infection. Anal intraepithelial neoplasia (AIN) is a potential precursor lesion of squamous cell carcinoma of the anus. Like its cervical counterpart, cervical intraepithelial neoplasia (CIN), AIN is causally linked to persistent infections with high-risk HPV types such as HPV16 or HPV18. As AIN and CIN share distinct biological similar-ities, AIN screenings analogous to Pap smear programs for CIN have been recommended in high-risk populations to reduce the incidence of anal carcinoma. These screenings include cytological analysis followed by high resolution anoscopy in case of anal dysplasia. Treatment guidelines for AIN are not yet available. Therapeutic strategies can be divided into topical (e.g. trichloroacetic acid, podophyllotoxin, imiquimod, photodynamic therapy) and ablative (e. g. surgical excision, laser ablation, infrared coagulation, electrocautery) measures. However, controlled studies on AIN treatment have not been performed. The impact of HPV vaccination on AIN development will also need to be assessed. Long-term follow-up of these patients is essential to gain more insight into the natural history of anogenital HPV infection in HIV-positive MSM. PMID- 18410394 TI - The use of a linear oscillating device in periodontal treatment: a review. AB - BACKGROUND: The Vector system is an ultrasonic instrument that was introduced in 1999, and treatment outcomes with Vector have been compared with those achieved with the "gold standard" of scaling and root planing with Gracey curettes as well as conventional ultrasonic scalers. The aim of this paper is to review the existing literature regarding the Vector method. MATERIAL AND METHODS: This review is based on a MEDLINE search of the use of Vector up to January 2008 (focused on in vitro findings, periodontal treatment, peri-implantitis therapy, clinical and microbiological data, and patient considerations). The MEDLINE search identified 128 papers, of which 18 were found to be relevant to this review. A hand search of the periodontal literature over the same period resulted in identification of a further 10 relevant papers. CONCLUSION: Vector used for treatment of chronic periodontitis results in clinical and microbiological outcomes comparable to those achieved by manual instrumentation and conventional ultrasonic instruments. Vector is less efficient when removing large masses of calculus, however, and cannot be recommended for the treatment of peri implantitis. Vector may be particularly useful in periodontal maintenance care as it is well tolerated by patients and results in less removal of cementum than other instruments. PMID- 18410395 TI - Vitamin D receptor polymorphism (-1056 Taq-I) interacts with smoking for the presence and progression of periodontitis. AB - AIM: The aim of this analysis was to investigate the relationship between a vitamin D receptor (VDR) polymorphism and the diagnosis and progression of periodontitis. MATERIAL AND METHODS: Data were derived from two different studies, including 231 subjects with healthy periodontium, 224 aggressive periodontitis and 79 chronic periodontitis (CP) patients in a case-control investigation. Sixty-one of these CP patients also took part in an observational study with a 1-year follow-up, in which progression of periodontitis was determined at the subject level. All 534 subjects provided a blood sample from which genomic DNA was extracted to study VDR -1056 TaqI polymorphism. RESULTS: The interaction between smoking and VDR polymorphism was associated with the diagnosis of periodontitis in Caucasians [p=0.001, odds ratio (OR)=1.33, 95% confidence intervals (CI)=1.12-1.57] and all subjects (p=0.033, OR=1.60, 95% CI=1.04-2.48). In the longitudinal study, subjects were divided into two clusters at 1 year according to the median number of progressing sites (Delta cumulative attachment loss >2 mm). Logistic regression analysis revealed that the interaction between VDR Taq-I polymorphism and smoking showed limited evidence of association with the "severe progression" cluster (p=0.033, OR=15.24, 95% CI=1.24 187.42). CONCLUSIONS: Vitamin D receptor Taq-I TT polymorphism was moderately associated with both the presence and the progression of periodontitis in smokers, while no association was detected in non-smoking individuals. VDR genetic factors may interact with smoking in the pathogenesis of periodontitis. PMID- 18410396 TI - Antibody levels to single bacteria or in combination evaluated against myocardial infarction. AB - BACKGROUND: Evidence is accumulating that oral bacteria are associated with myocardial infarctions (MI). We were interested in studying the differences in the association between single bacteria or bacteria in combination and the relation to C-reactive protein (CRP). MATERIAL AND METHODS: We examined the levels of antibodies against four major periodontal pathogens Porphyromonas gingivalis (PG), Aggregatibacter actinomycetemcomitans (AA), Tannerella forsythia (TF) and Treponema denticola (TD) and CRP in 548 men with a self-reported history of MI to 625 controls who took part in the Oslo II study in 2000. RESULTS: The mean levels of bacterial antibodies were higher for the cases than the controls, but not significant as standard deviations were large. The level of CRP was higher in the cases than the controls (p=0.010). Logistic regression analyses comparing the upper quartile value with the lower value of one of either four antibodies (anti-AA, anti-TF, anti-TD and anti-PG) were significantly associated (p=0.032) with MI. Equivalent analyses of either three bacteria showed significant associations for anti-AA, anti-TD and anti-PG (p=0.036) and anti-AA, anti-PG and anti-TF (p=0.040). CRP showed an increased relative risk with increasing quartile value; trend, p=0.016, but not in multivariate analysis including the oral antigens. CONCLUSIONS: No single bacterium but rather combinations were related to increasing relative risk for MI independent of known cardiovascular risk factors. PMID- 18410398 TI - Rapid assessment of risks to a mobile marine mammal in an ecosystem-scale marine protected area. AB - Ecosystem-scale networks of marine protected areas (MPAs) are important conservation tools, but their effectiveness is difficult to quantify in a time frame appropriate to species conservation because of uncertainties in the data available. The dugong (Dugong dugon) is a mobile marine species that occurs in shallow inshore waters of an ecosystem-scale network of MPAs (the Great Barrier Reef World Heritage Area [GBRWHA]). We developed a rapid approach to assess risk to dugongs in the region and evaluate options to ameliorate that risk. We used expert opinion and a Delphi technique to identify and rank 5 human factors with the potential to adversely affect dugongs and their sea grass habitats: netting, indigenous hunting, trawling, vessel traffic, and poor-quality terrestrial runoff. We then quantified and compared the distribution of these factors with a spatially explicit model of dugong distribution. We estimated that approximately 96% of habitat of high conservation value for dugongs in the GBRWHA is at low risk from human activities. Using a sensitivity analysis, we found that to decrease risk, commercial netting or indigenous hunting had to be reduced in remote areas and the effects of vessel traffic, terrestrial runoff, and commercial netting had to be reduced in urban areas. This approach enabled us to compare and rank risks so as to identify the most severe risks and locate specific sites that require further management attention. PMID- 18410397 TI - Clinical assessment of peri-implant tissues in patients with varying severity of chronic periodontitis. AB - PURPOSE: This retrospective study assessed the health of peri-implant tissues in patients with varying severity of chronic periodontitis. MATERIALS AND METHODS: Sixty-one subjects aged 44 to 70 years (median age 58 years) were recruited. Based on severity of periodontitis, 31 subjects were classified as having severe generalized chronic periodontitis, and the remaining 30 subjects had mild or no periodontitis. Social and medical histories were obtained from each patient. A comprehensive periodontal examination included: plaque index, gingival index, bleeding index, probing depth, clinical attachment level, and radiographic bone loss. Data were analyzed using Fisher's exact and chi-square tests for categorical variables, and t-test for continuous variables. RESULTS: There was a statistically significant greater loss of attachment (p < .05) around implants in the group with severe periodontitis compared to the no/mild periodontitis group. CONCLUSION: Because of the greater loss of clinical attachment around implants placed in patients with generalized severe chronic periodontitis, close monitoring of these patients is suggested to prevent both development of peri implantitis and recurrence of periodontal infection. PMID- 18410399 TI - Finding hope in the millennium ecosystem assessment. AB - Over the past quarter century, a new scientific activity has emerged: collective assessments by large numbers of scientists from different disciplines combining their expertise to better understand human interrelations with nature and to inform policy. The Millennium Ecosystem Assessment exceeded all such assessments before it in both the breadth of its coverage and the depth of its analysis of socioecological system dynamics. The findings are not encouraging. Nearly all ecosystems are being degraded and will continue to be degraded for decades to come even if policy changes are initiated now. For scientists participating in the assessment, the MA had another disconcerting aspect. It clearly shows that our fragmented, disciplinary knowledges cannot simply be combined to form an understanding of a whole complex system. Counterbalancing the despair of the findings and scientific difficulties of aggregating specialized knowledges, the MA demonstrated the potential of a deliberative democratic approach to grappling with complex problems. PMID- 18410400 TI - Economic valuation of biodiversity conservation: the meaning of numbers. AB - Recognition of the need to include economic criteria in the conservation policy decision-making process has encouraged the use of economic-valuation techniques. Nevertheless, whether it is possible to accurately assign economic values to biodiversity and if so what these values really represent is being debated. We reviewed 60 recent papers on economic valuation of biodiversity and carried out a meta-analysis of these studies to determine what factors affect willingness to pay for biodiversity conservation. We analyzed the internal variables of the contingent-valuation method (measure of benefits, vehicle of payment, elicitation format, or timing of payment) and anthropomorphic, anthropocentric and scientific factors. Funding allocation mostly favored the conservation of species with anthropomorphic and anthropocentric characteristics instead of considering scientific factors. We recommend researchers and policy makers contemplate economic valuations of biodiversity carefully, considering the inherent biases of the contingent-valuation method and the anthropomorphic and anthropocentric factors resulting from the public's attitude toward species. Because of the increasing trend of including economic considerations in conservation practices, we suggest that in the future interdisciplinary teams of ecologists, economists, and social scientists collaborate and conduct comparative analyses, such as we have done here. Use of the contingent-valuation method in biodiversity conservation policies can provide useful information about alternative conservation strategies if questionnaires are carefully constructed, respondents are sufficiently informed, and the underlying factors that influence willingness to pay are identified. PMID- 18410401 TI - Effects of forest fragmentation and habitat degradation on West African leaf litter frogs. AB - Habitat degradation alters the dynamics and composition of anuran assemblages in tropical forests. The effects of forest fragmentation on the composition of anuran assemblages are so far poorly known. We studied the joint influence of forest fragmentation and degradation on leaf-litter frogs. We specifically asked whether the processes structuring leaf-litter anuran assemblages in fragmented forests are the same as those in continuous forests. We analyzed anuran assemblages with respect to habitat characteristics, including fragmentation and degradation parameters. In comparison with continuous forests, species richness and diversity were lower and assemblage composition was altered in forest fragments. These changes seemed to be mainly caused by habitat degradation rather than forest fragmentation. Availability of aquatic sites for breeding, vegetation structure (including those variables indicating degradation), and leaf-litter cover had the most influence on the presence of single species. The comparatively small impact of fragmentation on anurans might be due to the location of the study area; it still possessed large tracts of continuous forest. These forest blocks may stabilize the regional rainforest climate and thus weaken the effects of fragmentation. PMID- 18410402 TI - Estimating sustainable bycatch rates for California sea lion populations in the Gulf of California. AB - Commercial and subsistence fisheries pressure is increasing in the Gulf of California, Mexico. One consequence often associated with high levels of fishing pressure is an increase in bycatch of marine mammals and birds. Fisheries bycatch has contributed to declines in several pinniped species and may be affecting the California sea lion (Zalophus californianus) population in the Gulf of California. We used data on fisheries and sea lion entanglement in gill nets to estimate current fishing pressure and fishing rates under which viable sea lion populations could be sustained at 11 breeding sites in the Gulf of California. We used 3 models to estimate sustainable bycatch rates: a simple population-growth model, a demographic model, and an estimate of the potential biological removal. All models were based on life history and census data collected for sea lions in the Gulf of California. We estimated the current level of fishing pressure and the acceptable level of fishing required to maintain viable sea lion populations as the number of fishing days (1 fisher/boat setting and retrieving 1 day's worth of nets) per year. Estimates of current fishing pressure ranged from 101 (0-405) fishing days around the Los Machos breeding site to 1887 (842-3140) around the Los Islotes rookery. To maintain viable sea lion populations at each site, the current level of fishing permissible could be augmented at some sites and should be reduced at other sites. For example, the area around San Esteban could support up to 1428 (935-2337) additional fishing days, whereas fishing around Lobos should be reduced by at least 165 days (107-268). Our results provide conservation practitioners with site-specific guidelines for maintaining sustainable sea lion populations and provide a method to estimate fishing pressure and sustainable bycatch rates that could be used for other marine mammals and birds. PMID- 18410403 TI - Increased apoptotic potential and dose-enhancing effect of gold nanoparticles in combination with single-dose clinical electron beams on tumor-bearing mice. AB - High atomic number material, such as gold, may be used in conjunction with radiation to provide dose enhancement in tumors. In the current study, we investigated the dose-enhancing effect and apoptotic potential of gold nanoparticles in combination with single-dose clinical electron beams on B16F10 melanoma tumor-bearing mice. We revealed that the accumulation of gold nanoparticles was detected inside B16F10 culture cells after 18 h of incubation, and moreover, the gold nanoparticles were shown to be colocalized with endoplasmic reticulum and Golgi apparatus in cells. Furthermore, gold nanoparticles radiosensitized melanoma cells in the colony formation assay (P = 0.02). Using a B16F10 tumor-bearing mouse model, we further demonstrated that gold nanoparticles in conjunction with ionizing radiation significantly retarded tumor growth and prolonged survival compared to the radiation alone controls (P < 0.05). Importantly, an increase of apoptotic signals was detected inside tumors in the combined treatment group (P < 0.05). Knowing that radiation-induced apoptosis has been considered a determinant of tumor responses to radiation therapy, and the length of tumor regrowth delay correlated with the extent of apoptosis after single-dose radiotherapy, these results may suggest the clinical potential of gold nanoparticles in improving the outcome of melanoma radiotherapy. PMID- 18410404 TI - Expression of RUNX3 in salivary adenoid cystic carcinoma: implications for tumor progression and prognosis. AB - Runt-related transcription factor-3 (RUNX3), being a tumor suppressor gene in gastric cancer, plays an important role in inhibiting cellular growth by participating in the transforming growth factor-beta-dependent apoptosis. The aim of this study was to determine the expression of RUNX3 in normal salivary glands and adenoid cystic carcinomas (ACCs), comparing the results with clinicopathological factors and patient survival. The quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis and Western blot analysis revealed the expression of RUNX3 both in normal salivary glands and ACCs. Nuclear and cytoplasmic immunoreactivities against RUNX3 in ductal luminal cells and acinous cells, but immunonegative in myoepithelial cells, were detected in normal salivary glands. In ACC, the RUNX3 immunostaining was shown in the cytoplasm of tumor cells; however, no nuclear location of RUNX3 was found. Lower RUNX3 expression showed significant correlation to distant metastasis and histological growth pattern (P = 0.009 and P = 0.025, respectively). On univariate analysis, low level of RUNX3 immunolabeling (P = 0.012), stage T4 (P = 0.017), lymph node involvement (P = 0.007), and distant metastasis (P < 0.001) were significantly associated with decreased overall survival. Multivariate analysis showed only distant metastasis had an independent prognostic effect on overall survival (P = 0.043). Our results demonstrate the expression of RUNX3 in normal salivary glands and salivary ACCs. The low level of RUNX3 protein in salivary ACCs might play a pivotal role in tumor progression and have prognostic values in ACCs. PMID- 18410405 TI - Gli1 contributes to the invasiveness of pancreatic cancer through matrix metalloproteinase-9 activation. AB - The hedgehog (Hh) signaling pathway has been reported to be associated with the growth of pancreatic cancer, but its role in the invasive phenotype is poorly understood. Therefore, we investigated the role of the Hh pathway in pancreatic cancer cell invasiveness using a Matrigel invasion assay. Blockade of the Hh pathway by cyclopamine inhibited pancreatic cancer cell invasion in association with a decreased expression of matrix metalloproteinase (MMP)-9. By contrast, activation of the Hh pathway by the addition of exogenous Sonic hedgehog increased cell invasion and MMP-9 expression. Stable transfection of pancreatic cancer cells with Gli1 increased their invasiveness, which was associated with activation of MMP-9. We also showed that inhibition of MMP-9 by small interfering RNA blocked the increased invasiveness of Gli1-transfected cells. Furthermore, inhibition of Gli1 by small interfering RNA suppressed the invasiveness and MMP-9 expression of pancreatic cancer cells. Taken together, these findings suggest that members of the Hh pathway, especially Gli1, play an important role in the invasiveness of pancreatic cancer cells through the regulation of MMP-9 expression. PMID- 18410406 TI - Polyketides in insects: ecological role of these widespread chemicals and evolutionary aspects of their biogenesis. AB - Polyketides are known to be used by insects for pheromone communication and defence against enemies. Although in microorganisms (fungi, bacteria) and plants polyketide biogenesis is known to be catalysed by polyketide synthases (PKS), no insect PKS involved in biosynthesis of pheromones or defensive compounds have yet been found. Polyketides detected in insects may also be biosynthesized by endosymbionts. From a chemical perspective, polyketide biogenesis involves the formation of a polyketide chain using carboxylic acids as precursors. Fatty acid biosynthesis also requires carboxylic acids as precursors, but utilizes fatty acid synthases (FAS) to catalyse this process. In the present review, studies of the biosynthesis of insect polyketides applying labelled carboxylic acids as precursors are outlined to exemplify chemical approaches used to elucidate insect polyketide formation. However, since compounds biosynthesised by FAS may use the same precursors, it still remains unclear whether the structures that are formed from e.g. acetate chains (acetogenins) or propanoate chains (propanogenins) are PKS or FAS products. A critical comparison of PKS and FAS architectures and activities supports the hypothesis of a common evolutionary origin of these enzyme complexes and highlights why PKS can catalyse the biosynthesis of much more complex products than can FAS. Finally, we summarise knowledge which might assist researchers in designing approaches for the detection of insect PKS genes. PMID- 18410407 TI - Cerebral amyloid angiopathy and parenchymal amyloid deposition in transgenic mice expressing the Danish mutant form of human BRI2. AB - Familial Danish dementia (FDD) is an autosomal dominant neurodegenerative disease clinically characterized by the presence of cataracts, hearing impairment, cerebellar ataxia and dementia. Neuropathologically, FDD is characterized by the presence of widespread cerebral amyloid angiopathy (CAA), parenchymal amyloid deposition and neurofibrillary tangles. FDD is caused by a 10-nucleotide duplication-insertion in the BRI(2) gene that generates a larger-than-normal precursor protein, of which the Danish amyloid subunit (ADan) comprises the last 34 amino acids. Here, we describe a transgenic mouse model for FDD (Tg-FDD) in which the mouse Prnp (prion protein) promoter drives the expression of the Danish mutant form of human BRI(2). The main neuropathological findings in Tg-FDD mice are the presence of widespread CAA and parenchymal deposition of ADan. In addition, we observe the presence of amyloid-associated gliosis, an inflammatory response and deposition of oligomeric ADan. As the animals aged, they showed abnormal grooming behavior, an arched back, and walked with a wide-based gait and shorter steps. This mouse model may give insights on the pathogenesis of FDD and will prove useful for the development of therapeutics. Moreover, the study of Tg FDD mice may offer new insights into the role of amyloid in neurodegeneration in other disorders, including Alzheimer disease. PMID- 18410408 TI - Combining etanercept and acitretin in the therapy of chronic plaque psoriasis: a 24-week, randomized, controlled, investigator-blinded pilot trial. AB - BACKGROUND: Combination treatments may increase efficacy while reducing dosages and side-effects of individual agents. No randomized controlled trials have been published combining biologics with conventional agents for psoriasis. OBJECTIVES: To investigate the efficacy and safety of the association of acitretin and etanercept in the treatment of moderate to severe chronic plaque psoriasis. METHODS: A 24-week, randomized, controlled, investigator-blinded pilot trial was conducted. Sixty adult patients with moderate to severe chronic plaque psoriasis were randomized into three groups to receive etanercept 25 mg twice weekly subcutaneously, oral acitretin 0.4 mg kg(-1) daily or etanercept 25 mg once weekly plus acitretin 0.4 mg kg(-1) daily. The primary end point was a 75% or greater improvement in Psoriasis Area and Severity Index (PASI) from baseline (PASI 75) at week 24. RESULTS: At week 24, PASI 75 response was achieved by 10 of 22 patients in the etanercept group (45%), six of 20 in the acitretin group (30%) and eight of 18 (44%) in the group treated with etanercept plus acitretin (P = 0.001 for both etanercept groups compared with acitretin alone). A 50% or greater improvement from baseline in PASI was achieved by 15 of 22 (68%), 10 of 20 (50%) and 12 of 18 (67%) patients, respectively (P = 0.001). The safety profiles of the three groups were similar. CONCLUSIONS: A combined therapeutic regimen with etanercept 25 mg once weekly and acitretin 0.4 mg kg(-1) daily is as effective as etanercept 25 mg twice weekly, and more effective than acitretin alone. Although larger studies are needed to confirm these results, the etanercept/acitretin association could offer several advantages in the therapy of moderate to severe chronic plaque psoriasis. PMID- 18410409 TI - Detection of human papillomavirus type 56 in Bowen's disease involving the nail matrix. AB - BACKGROUND: As Bowen's disease of the nail apparatus is quite rare, there have been only a few reports on the prevalence of human papillomavirus (HPV) infection in this condition. OBJECTIVES: The purpose of this study was to clarify the association of HPV with this disease involving the nail apparatus. METHODS: Five patients with Bowen's disease of the nail apparatus were investigated clinically, virologically and histologically. Total DNAs extracted from excised skin lesions were analysed using polymerase chain reaction (PCR) for the presence of HPV DNA and the amplified products were subjected to DNA sequence analyses. Histological localization of HPV DNA was examined by in situ hybridization. RESULTS: In three of five patients, HPV was detected by PCR amplification, and subsequent sequence analyses of the PCR products showed the sequences of HPV type 56. A common clinical feature of the three HPV-positive patients was longitudinal melanonychia. In contrast, the two HPV-negative patients presented with a convex nail deformity and a periungual ulcerative lesion. In two of three positive cases, there was a silent point mutation in the L1 gene of each HPV. In the remaining one case, the nucleotide sequence was consistent with the consensus sequence of HPV 56. Sequence analyses of the E6 gene revealed the infection of different variants of HPV 56 among the three cases. The viral genomes were located in keratinocyte nuclei upon in situ hybridization. CONCLUSIONS: HPV 56 may be involved in the carcinogenesis of Bowen's disease affecting the nail matrix with longitudinal pigmentation. PMID- 18410410 TI - Ultraviolet C inactivation of dermatophytes: implications for treatment of onychomycosis. AB - BACKGROUND: Onychomycosis responds to systemic antifungals and sometimes to topical lacquers, but alternative treatments are desirable. Topical application of germicidal ultraviolet (UV) C radiation may be an acceptable and effective therapy for infected nails. OBJECTIVES: To test the ability of UVC to inactivate dermatophyte suspensions in vitro and to sterilize a novel ex vivo model of nail infection. METHODS: Trichophyton rubrum, T. mentagrophytes, Epidermophyton floccosum and Microsporum canis suspensions were irradiated with UVC (254 nm) at a radiant exposure of 120 mJ cm(-2) and surviving colony-forming units quantified. T. rubrum infecting porcine hoof slices and human toenail clippings was irradiated with UVC at radiant exposures of 36-864 J cm(-2). RESULTS: In vitro studies showed that 3-5 logs of cell inactivation in dermatophyte suspensions were produced with 120 mJ cm(-2) UVC irradiation. Depending on factors such as the thickness and infectious burden of the ex vivo cultures, the radiant exposure of UVC needed for complete sterilization was usually in the order of tens to hundreds of J cm(-2). Resistance of T. rubrum to UVC irradiation did not increase after five cycles of subtotal inactivation in vitro. CONCLUSIONS: UVC irradiation may be a less invasive treatment option for onychomycosis, when the appropriate consideration is given to safety. PMID- 18410411 TI - Chanarin-Dorfman syndrome caused by a novel splice site mutation in ABHD5. PMID- 18410412 TI - Extremely low frequency electromagnetic field enhances human keratinocyte cell growth and decreases proinflammatory chemokine production. AB - BACKGROUND: Proliferation and differentiation of keratinocytes are central processes in tissue regeneration after injury. Chemokines, produced by a wide range of cell types including keratinocytes, play a regulatory role in inflammatory skin diseases. Several studies have shown that an electromagnetic field (EMF) can influence both inflammatory processes and repair mechanisms including wound healing on different tissue models. OBJECTIVES: To elucidate the effect of extremely low frequency EMF (ELF-EMF) on keratinocyte proliferation and production of chemokines [RANTES, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1 alpha and interleukin (IL)-8] in order to evaluate a potential therapeutic use of magnetic fields. METHODS: The human keratinocyte cell line HaCaT was exposed at 1 mT, 50 Hz for different lengths of time and compared with unexposed control cells. Cell growth and viability were evaluated at different exposure times by cell count and trypan blue exclusion. Chemokine production and expression were analysed by enzyme-linked immunosorbent assay (ELISA) and by real-time polymerase chain reaction. Total NF-kappaB p65 was quantified by ELISA. RESULTS: Significantly increased growth rates were observed after 48 h of EMF exposure as compared with control cells, while no difference in cell viabilities were detected. Gene expression and release of RANTES, MCP-1, MIP 1 alpha and IL-8 were significantly reduced after 72 h of exposure. NF-kappaB levels became almost undetectable after only 1 h of EMF exposure, and were inversely correlated with cell density. CONCLUSIONS: Our results show that ELF EMF modulates chemokine production and keratinocyte growth through inhibition of the NF-kappaB signalling pathway and thus may inhibit inflammatory processes. ELF EMF could represent an additional therapeutic approach in the treatment of skin injury. PMID- 18410413 TI - Updates from the British Association of Dermatologists 87 th annual meeting, 10 13 July 2007, Birmingham, U.K. AB - This is a synopsis of the main research and clinical findings presented at the British Association of Dermatologists meeting held during 10-13 July 2007 in Birmingham, U.K. The conference highlighted the recent biological, epidemiological and therapeutic advances that have been made recently in the field of dermatology. The authors focus on the more important advances or summaries of findings, but this is not meant as a substitute for reading the conference proceedings and related references quoted in this article. PMID- 18410414 TI - MUM1 expression in cutaneous CD30+ lymphoproliferative disorders: a valuable tool for the distinction between lymphomatoid papulosis and primary cutaneous anaplastic large-cell lymphoma. AB - BACKGROUND: Primary cutaneous CD30+ lymphoproliferative disorders include lymphomatoid papulosis (LyP) and primary cutaneous CD30+ anaplastic large T-cell lymphoma (ALCL). Because of overlapping histological features, it is impossible to distinguish ALCL from LyP on histological grounds. MUM1 (Multiple Myeloma oncogene 1) is expressed in systemic ALCL and classical Hodgkin lymphoma. MUM1 expression has not been studied in detail in CD30+ lymphoproliferative disorders. OBJECTIVES: To examine the expression of MUM1 in CD30+ lymphoproliferative disorders and to assess its value as a diagnostic marker. METHODS: Thirty-one formalin-fixed paraffin-embedded specimens of LyP (n = 15), primary cutaneous ALCL (n = 10), secondary cutaneous infiltrates of systemic ALCL (n = 4) and secondary cutaneous Hodgkin lymphoma (n = 2) were analysed by immunohistochemistry with a monoclonal antibody against MUM1. RESULTS: Positive staining for MUM1 was observed in 13 cases of LyP (87%), two cases of primary cutaneous ALCL (20%), four cases of secondary cutaneous ALCL (100%) and two cases of secondary cutaneous Hodgkin lymphoma (100%). In 11 of 13 LyP cases (85%), MUM1 was displayed by the majority, i.e. 50-90%, of the tumour cells. In contrast to LyP and secondary cutaneous ALCL, only two cases of primary cutaneous ALCL (20%) harboured MUM1-positive tumour cells. There was a statistically significant difference in the expression of MUM1 between LyP and primary cutaneous ALCL (P = 0.002) and between primary cutaneous ALCL and secondary cutaneous ALCL (P = 0.015). CONCLUSIONS: MUM1 expression is a valuable tool for the distinction of LyP and ALCL and thus represents a novel adjunctive diagnostic marker in CD30+ lymphoproliferative disorders. PMID- 18410416 TI - Association between beta-blockers, other antihypertensive drugs and psoriasis: population-based case-control study. AB - BACKGROUND: Several case reports have associated use of beta-blockers with an increased risk of psoriasis or psoriasiform drug eruptions. OBJECTIVES: To study the association between use of beta-blockers and other antihypertensive drugs and the risk of developing a first-time diagnosis of psoriasis. METHODS: We conducted a case-control analysis on the U.K.-based General Practice Research Database. We identified cases with an incident psoriasis diagnosis between 1994 and 2005 and matched them to one control patient on age, sex, general practice, calendar time (same index date) and years of history in the database. Conditional logistic regression was used to estimate adjusted odds ratios (ORs) with 95% confidence intervals (CIs) of developing a first-time psoriasis diagnosis in relation to previous exposure to antihypertensive drugs, stratified by exposure timing (current vs. past use) and exposure duration based on the number of prescriptions. RESULTS: The study encompassed 36 702 cases with a first-time psoriasis diagnosis and the same number of matched controls. Adjusted ORs for current use of 1-4, 5-19 or >or= 20 prescriptions for beta-blockers, as compared with nonuse, were 0.93 (95% CI 0.76-1.13), 1.10 (95% CI 0.97-1.24), and 1.10 (95% CI 1.01-1.20), respectively. The risk estimates for current use of other antihypertensives at any exposure duration were all close to 1.0. CONCLUSIONS: This large population-based case-control analysis does not support the current proposition that beta-blocker use is associated with an increased risk of psoriasis, nor did we find evidence for a substantially altered psoriasis risk for other antihypertensive drugs. PMID- 18410417 TI - Interstitial granulomatous drug reaction following intravenous ganciclovir. PMID- 18410415 TI - Polymorphisms in the interleukin 13 and GATA binding protein 3 genes and the development of eczema during childhood. AB - BACKGROUND: Atopic eczema is characterized by Th2-dominant immunity with the cytokine interleukin 13 and the transcription factor GATA binding protein 3 playing a critical role. OBJECTIVES: We assessed the association of polymorphisms in the IL13 and GATA3 genes with childhood eczema. METHODS: A birth cohort (n = 1456) was established on the Isle of Wight in 1989 and followed at the ages of 1 (n = 1167), 2 (n = 1174), 4 (n = 1218) and 10 years (n = 1373) to determine the prevalence of allergic disease including eczema. At 4 and 10 years, skin prick testing was performed. Whole blood samples (n = 923) were obtained at the 10-year assessment, stored frozen, and genotyped. Five polymorphisms from IL13 and seven from GATA3 were genotyped for this analysis. Repeated measurement analyses were conducted for the occurrence of eczema at ages 1, 2, 4 and 10 years. All analyses were adjusted for maternal and paternal eczema, low birth weight (< 2500 g), breastfeeding >or= 3 months and age. RESULTS: IL13 was not associated with childhood eczema. For GATA3, the single nucleotide polymorphism (SNP) rs2275806 (promoter region) showed an increased odds ratio for atopic eczema independent of whether the comparison group had a positive skin prick test. The SNP rs444762 (intron 3 region) was associated with atopic eczema in comparison with children without eczema. The increased relative risks remained significant after adjustment for multiple testing only for rs2275806 (P < 0.05). CONCLUSIONS: A SNP in GATA3 is associated with atopic eczema. This finding highlights the importance of GATA3 as an immune-modulating gene in atopic eczema. PMID- 18410418 TI - Unilateral segmental acneiform naevus: a model disorder towards understanding fibroblast growth factor receptor 2 function in acne? PMID- 18410419 TI - Chronic hepatitis B reactivation and systemic glucocorticosteroid therapy. PMID- 18410420 TI - Topical 5-fluorouracil in Darier disease. PMID- 18410421 TI - The things our patients do to avoid operations (do-it-yourself Mohs' surgery)! PMID- 18410422 TI - Transforming growth factor-beta1-induced CD4+CD25+ regulatory T cells in vitro reverse and prevent a murine lupus-like syndrome of chronic graft-versus-host disease. AB - BACKGROUND: The suppressive mechanism of CD4+CD25+ regulatory T cells (Tregs) is poorly understood. It is also not known how to obtain enough peripheral Tregs, and how to make them effective in ameliorating a murine lupus-like syndrome of chronic graft-versus-host disease (cGVHD). OBJECTIVES: To confirm the contribution of transforming growth factor (TGF)-beta1 in the function of CD4+CD25+ Tregs in vitro, and to identify in vivo suppressive effects of different Tregs generated through TGF-beta1. METHODS: Suppressive effects of freshly isolated CD4+CD25+ Tregs, TGF-beta1-expanded CD4+CD25+ Tregs (eTregs) and TGF-beta1-induced CD4+CD25+ Tregs (iTregs) in vitro were assessed. Reverse transcription-polymerase chain reaction was used to detect Foxp3. The respective roles that different Tregs might play in controlling murine lupus-like syndrome of cGVHD were analysed. RESULTS: TGF-beta1 was necessary for expanding the existing CD4+CD25+ Tregs in vitro, as well as converting peripheral CD4+CD25- T cells to CD4+CD25+ Tregs through upregulating CD25 and Foxp3. These eTregs and iTregs had a suppressive effect similar to that of freshly isolated CD4+CD25+ Tregs. The inhibitory function of iTregs could be partially blocked by anti-TGF beta1. Importantly, it was revealed for the first time that both eTregs and iTregs had an inhibitory effect on reversing the morbidity of mice that had already developed anti-dsDNA, and iTregs gave more suppression than eTregs. Besides, iTregs could prevent the onset and slow the progress of disease in a significantly dose-dependent manner. CONCLUSIONS: Results indicate that TGF-beta1 signalling is required to maintain the suppression of CD4+CD25+ Tregs in vitro and in vivo. Together, this study suggests a possible therapeutic role for iTregs in the treatment of murine lupus-like syndrome of cGVHD. PMID- 18410424 TI - A novel frameshift mutation in the KIND1 gene in Turkish siblings with Kindler syndrome. PMID- 18410425 TI - Rapidly involuting congenital haemangioma associated with transient thrombocytopenia and coagulopathy: a case series. AB - Rapidly involuting congenital haemangioma (RICH) may present with thrombocytopenia, low fibrinogen and elevated fibrin degradation products and D dimers. Such complications have rarely been reported. We wished to define the clinical characteristics of the thrombocytopenia and coagulopathy associated with RICH, to emphasize the transient nature of this haematological complication and to distinguish these abnormalities from true Kasabach-Merritt phenomenon (KMP). We present a case series of seven patients with large RICH who presented with thrombocytopenia and coagulopathy during the first week of life. Clinical and haematological characteristics were recorded retrospectively. Two of the patients were treated with embolization due to early signs of high-output cardiac failure; four patients received oral corticosteroids in the range of 2 mg kg(-1) daily; one patient did not receive any treatment in the neonatal period, although the tumour was excised at 6 months of age. Two patients with platelet counts lower than 10 x 10(9) L(-1) received a platelet transfusion. There were no bleeding complications and only one patient presented with petechiae. In all seven patients, platelet counts started to increase at > 2 weeks of age and the coagulopathy resolved. We conclude that RICH may present with thrombocytopenia and coagulopathy similar to mild KMP early in the neonatal period. However, in contrast to true KMP, these abnormal laboratory findings are self-limited and are usually not complicated by bleeding problems. PMID- 18410426 TI - Stromal CD10 expression is correlated with invasiveness and proliferation of extramammary Paget disease. PMID- 18410427 TI - Laparoscopic sentinel node dissection for prostate carcinoma: technical and anatomical observations. AB - OBJECTIVE: To report experience with sentinel node (SN) lymphadenectomy which allows an assessment of the exact location of radioactive and of tumour-bearing lymph nodes, and evaluate differences in timing of the scintigraphy and surgery. PATIENTS AND METHODS: The study included 35 patients who opted for external beam radiation therapy for prostate carcinoma of intermediate or poor prognosis. Agreement was reached between the participating urologists and the physicians of the nuclear medicine department on the definition of the relevant anatomical areas. The time between a transrectal intraprostatic injection with the radioactive nanocolloid and the laparoscopic SN procedure varied from 5 h to 26 h. Scintigrams were merged with the computed tomography scans until combined methods became available. A laparoscopic gamma-probe was used to identification the SNs, and an extensive laparoscopic node dissection undertaken in the same procedure. Lymph nodes were submitted to the pathologist in such a way that their exact location could be reconstructed. After surgery a graphic report was produced showing the exact location of the lymph nodes. RESULTS: Of the 35 patients 40% were node positive; a mean of 13.5 nodes were resected, and there were no false-negative results. The location of the vast majority of the tumour positive SNs was around the bifurcation of the external and internal iliac artery, and so involved nodes from the internal iliac, external iliac, communis and obturator basins. Of the six SNs outside the extended node dissection area, two were positive but only one of them exclusively so (lateral to the external iliac artery). The scintigrams did not change after 4 h, and the operation should be done within 24 h to have sufficient radioactivity in the nodes to be detected by the probe. There were eight complications (23%) but only one could be attributed to the SN procedure; the others were thought to be related to the extended laparoscopic node dissection. CONCLUSION: The laparoscopic SN procedure is a reliable tool for diagnosing prostate cancer-bearing lymph nodes, but the extended laparoscopic node dissection has, in this series and others, too many complications for it to be attractive for diagnostic purposes. The SN procedure makes an extended node dissection unnecessary in most patients. PMID- 18410428 TI - The role of lymph-node dissection in the treatment of upper urinary tract cancer: a multi-institutional study. AB - OBJECTIVES: To determine the role of lymph-node (LN) dissection in patients undergoing surgery for upper urinary tract (UUT) cancer. PATIENTS AND METHODS: We reviewed the clinicopathological data from 312 patients with UUT cancer treated predominantly by nephroureterectomy. The relationship between clinical characteristics and cancer-specific survival (CSS) was analysed, focusing on node related information. RESULTS: In all, 166 patients had LN dissection while 146 did not (pNx). Multivariate analysis showed that T stage, grade and pN status were significant variables for CSS. The difference in survival between the pN0 and pNx groups remained significant in a multivariate analysis. The median (range) number of LNs removed was 6 (1-65). There was no significant difference in CSS between the 72 patients with fewer than six LNs removed and the 78 with six or more removed. CONCLUSIONS: LN dissection is important for postoperative stratification of patients with UUT cancer because node-positive disease was one of the variables with a significant adverse effect on survival. In addition, the significant difference in survival between the pN0 and pNx groups might indicate a therapeutic benefit of LN dissection, although removing more LNs did not uniformly increase the probability of CSS. PMID- 18410429 TI - Outcome of pregnancy in women with a history of vesico-ureteric reflux. AB - OBJECTIVES: To review the evidence relating to the outcome of pregnancy in women with vesico-ureteric reflux (VUR) or a previous history of VUR and to identify the factors contributing to morbidity in pregnancy, with particular emphasis on the role of renal scarring. METHODS: Searches were carried out in Medline, Pubmed and MD Consult using various combinations of the keywords including: vesicoureteral reflux and pregnancy, maternal vesicoureteral reflux, vesicoureteral reflux in adulthood, reflux nephropathy and pregnancy. All data quoted in this review are from original articles. RESULTS: The published studies showed that women with VUR that was not associated with renal scarring had no increase in the incidence of gestational hypertension, pre-eclampsia or fetal morbidity, regardless of whether their VUR was diagnosed in childhood or adulthood. However, women with VUR and normal kidneys did have higher incidence of urinary tract infection during pregnancy, which was not modified by ureteric re-implantation. Renal scarring was the primary risk factor for morbidity during pregnancy and this risk was independent of the presence or absence of VUR at the time of pregnancy. CONCLUSION: The evidence does not support the practice of correcting low-grade VUR in girls with unscarred kidneys because this will reduce their risk of complications in pregnancy. The presence of renal scarring rather than the presence or absence of reflux is the principal determinant of morbidity during pregnancy. PMID- 18410430 TI - Management of renal cancer in the tyrosine kinase inhibitor era: a view from 3 years on. AB - In the last 3 years there has been a dramatic increase in the treatment options for patients with metastatic renal cancer. In addition to the cytokines interferon and interleukin 2, recently approved agents include sorafenib, sunitinib, temsirolimus and bevacizumab. A plethora of agents that are likely to have clinical activity are currently in the development 'pipeline'. This brief review is intended to overview recent developments, and to identify advances that are likely to influence treatment decisions. PMID- 18410431 TI - Phosphodiesterase type 5 inhibitors in the management of erectile dysfunction secondary to treatments for prostate cancer: findings from a Cochrane systematic review. PMID- 18410432 TI - Genetic polymorphism and pathogenesis of benign prostatic hyperplasia. PMID- 18410433 TI - How long is too long? The effect of the duration of anaesthesia on the incidence of non-urological complications after surgery. AB - OBJECTIVE: To examine the association between the duration of anaesthesia and non urological complication (NUC) rates after surgery, as the increasing complexity of minimally invasive and laparoscopic procedures in urology has resulted in longer surgery and anaesthesia, and information on the effect of this on NUC rates is limited. PATIENTS AND METHODS: We identified 2196 patients who had open radical nephrectomy or nephron-sparing surgery at our institution between 1989 and 2002. Patients were subdivided into groups I, II, and III according to the duration of general anaesthesia (<4, 4-6 and > or =6 h, respectively). NUCs after surgery, and mortality during and after surgery, were evaluated. RESULTS: There were 1340, 723, and 133 patients in groups I, II and III, respectively. The incidences of any NUC were 3.1%, 5.8% and 13.5%, respectively. The odds ratios for the likelihood of a NUC were 1.91 (P = 0.004) and 4.84 (P < 0.001) for groups II and III, respectively. These differences remained significant even after adjusting for patient and tumour characteristics. Perioperative mortality was highest in group III, at 2.3%, vs 0.4% in groups I and II. CONCLUSIONS: Longer anaesthesia is associated with an increase in the incidence of perioperative complications and mortality, especially when the duration of anaesthesia is >6 h. This increase appears to be independent of patient's preoperative health status, tumour extent and blood loss. PMID- 18410434 TI - Type 4 phosphodiesterase inhibitor suppresses experimental bladder inflammation. AB - OBJECTIVE: To evaluate the effects of orally administered YM976, a specific inhibitor of type 4 phosphodiesterase (PDE4), on bladder activity in a rat model with hydrochloric acid (HCl)-induced cystitis (IC), hypothesizing that a PDE4 inhibitor might suppress bladder overactivity and bladder pain responses in bladder-hypersensitive disorders such as IC. MATERIALS AND METHODS: Wistar rats with HCl-induced IC were treated with YM976 or vehicle and their voiding observed and assessed by cystometry. The severity of bladder inflammation (BI) was quantified using the BI index (BII), which comprises three factors (oedema, leukocyte infiltration and haemorrhage). Nociceptive neural activity was also examined using an immunohistochemical study of spinal c-fos expression. RESULTS: YM976 significantly reduced the number of voids, and the volume per void was significantly higher than in control (vehicle) group. Cystometry showed a significant increase in bladder capacity, voided volume and voiding efficiency, and a decrease in the amplitude of voiding pressure in rats treated with YM976. All BII scores were significantly lower in the YM976 than in the control group. c fos expression in the spine was less in the YM976 than in the control group. CONCLUSIONS: Oral administration of YM976 significantly improved the voiding behaviour and histological damage in rats with IC induced by HCl. These results indicate that PDE4 inhibitor might be effective in relieving bladder symptoms with IC. PMID- 18410435 TI - Renal tumour surgery in elderly patients. AB - OBJECTIVE: To analyse morbidity, renal function and oncological outcome in patients aged >or=80 years who had surgery for renal tumours, as in the elderly such surgery is controversial in relation to life-expectancy and other causes of death. PATIENTS AND METHODS: Between 1990 and 2006, in our institution 1625 patients had surgery to treat solid renal tumours suspected to be renal cell carcinoma (RCC); 62 (4%) were aged >or=80 years (mean 82.5), and 73% of these elderly patients had radical nephrectomy (RN) and 27% nephron-sparing surgery (NSS). RESULTS: The median (range) follow-up was 3.1 (0.2-14.1) years (89% of the patients). There was no perioperative mortality. There were only minor complications in 47% of patients, most (34%) being temporary increases in serum creatinine level. Histopathologically, 10% of the 62 patients had benign lesions and 90% had RCC. Of the 56 patients with RCC, the stage was pT1a in 34%, pT1b in 25%, pT2 in 5% and pT3 in 36%. For those treated with RN the median (range) serum creatinine level before and after RN was 1.0 (0.7-1.8) and 1.4 (1.0-2.8) mg/dL (P < 0.05), and for those treated with NSS were 1.1 (0.7-4.4) and 1.2 (0.7-4.8) mg/dL (not significant), respectively. The 5-year overall survival was 68% and the cancer-specific survival was 85%. CONCLUSIONS: Surgery for renal tumours is safe in elderly patients, with a low perioperative morbidity and a good overall survival rate. Patients should be selected carefully according to comorbidities, biological age and social support. PMID- 18410436 TI - Urinary fibronectin as a predictor of a residual tumour load after transurethral resection of bladder transitional cell carcinoma. AB - OBJECTIVE: To determine whether the level of urinary fibronectin predicts the residual tumour load after transurethral resection (TUR) of bladder transitional cell carcinoma (TCC). PATIENTS AND METHODS: Urine samples were collected from 167 consecutive patients with suspected bladder cancer admitted for TUR. Samples were taken both before and after surgery. Bladder tumour fibronectin (BTF) was analysed using a solid-phase chemiluminescent immunometric test. Creatinine in urine was also determined and the BTF/creatinine ratio calculated. RESULTS: Patients were divided into a control group of 41 whose previous diagnosis was negative for BT and another of 126 with a positive diagnosis for BT, which was further subdivided into those with and without residual tumour, according to findings from specimens obtained during the second procedure (repeat TUR or cystectomy). After the second procedure, 68 patients (56%) had no residual tumour, whereas 54 (44%) did. Four patients with BT who did not have the second procedure were excluded from the study. The median BTF and BTF/creatinine value in the control group was 33.2 microg/L and 51.4 microg/g, respectively, before the first TUR, and 29.6 microg/L and 46.7 microg/g, respectively, after the first TUR. There were no statistically significant changes in BTF and BTF/creatinine ratio (P = 0.61 and 0.79, respectively). In the group with TCC, the BTF decreased from 211.9 to 97.3 microg/L (P = 0.02) and the BTF/creatinine ratio from 281.6 to 146.5 microg/g (P = 0.009) for those with residual tumour, while it decreased from 195.1 to 34.0 microg/L (P = 0.007) and the BTF/creatinine ratio decreased from 249.1 to 53.7 microg/g (P = 0.003) for those with no residual tumour. After initial TUR, the patients with residual tumour had significantly greater levels of BTF and BTF/creatinine than did those with no residual tumour (P = 0.004 and 0.006, respectively). The receiver operating characteristic curves showed an optimum threshold of 67.8 microg/L and 81.3 microg/g for BTF and the BTF/creatinine in detecting residual tumour, respectively, with a sensitivity of 91.4% and 89.0%, respectively, and a specificity of 87.8% and 85.6%, respectively. CONCLUSION: Urinary fibronectin, in addition to being one of the best markers for diagnosing bladder carcinoma, can be used to determine the presence of residual tumour load after TUR of bladder TCC. PMID- 18410437 TI - Cystectomy in the elderly: does the survival benefit in younger patients translate to the octogenarians? AB - OBJECTIVE: To determine whether the survival benefit achieved with radical cystectomy (RC, the reference standard for treating muscle-invasive bladder cancer) in younger patients justifies its use in octogenarians. PATIENTS AND METHODS: We used the Surveillance Epidemiology and End Results data of the National Cancer Institute and identified 10 807 patients from 1992-2004 who were diagnosed with muscle-invasive bladder cancer, and were treated with either RC or radiotherapy. The data were analysed for age, gender, race, extent of lymphadenectomy and cause of death. We stratified the patients by age groups (<60, 60-69, 70-79 and >79 years), and used Kaplan-Meier survival analysis to compare treatment strategies by age group. RESULTS: In all, 8034 patients had RC and 2773 radiotherapy; RC was the primary method of treatment in all age groups except for octogenarians. Those who had RC had a sizeable overall survival advantage in all age groups, except for the octogenarians (18 vs 15 months). This small survival advantage improved only slightly (23 vs 15 months) when excluding patients having nodal or distant metastasis. The octogenarians who have RC with a limited pelvic lymph node dissection or RC alone receive little (16 vs 15 months) or no survival benefit. However, cancer-specific survival was significantly higher in those who had RC, including octogenarians. CONCLUSIONS: Octogenarians have some benefit to cancer-specific survival from RC if it includes a standard lymphadenectomy. The issue is how to better select the patients, as the overall survival advantage in these patients over radiotherapy is negligible. PMID- 18410438 TI - Changes to the contractile function of ureter smooth muscle after partial infravesical obstruction in fetal sheep. AB - OBJECTIVE: To measure spontaneous contractile activity, and the responses to agonists using in vitro preparations of sheep ureter after a period of bladder outlet obstruction (BOO) initiated at mid-gestation. MATERIAL AND METHODS: Date mated Romney Marsh ewes, bearing fetuses of 70-75 days of gestation (term in this breed is 145 days) were used. Five fetuses underwent urachal obstruction and partial urethral constriction for 30 days. Six fetuses at the same gestational time underwent a sham operation (control group). Small strips of mid-ureter were cut in the longitudinal axis, and in the cross-sectional (transverse) plane of the ureter. Spontaneous contractile activity and the response to carbachol, high K(+) (120 mM) solution and alpha,beta-methylene ATP (ABMA) were characterized by measuring the magnitude of evoked responses and the magnitude and frequency of spontaneous activity. RESULTS: The ureters from fetuses with BOO were significantly larger in diameter and had expanded lumens. The proportion of smooth muscle was not significantly different between the BOO and control groups. Spontaneous contractile activity in all preparations was resistant to atropine and the neurotoxin, tetrodotoxin. With transverse sections, the magnitude of spontaneous contractions was smaller in the BOO group, but the frequency was greater. The response to carbachol was also smaller in the BOO group, but the response to high-K(+) solution was similar to that of the control group. ABMA did not generate a response in any preparation. With longitudinal ureteric preparations, spontaneous or agonist-induced activity was negligible in the control group, while preparations from the BOO sheep had spontaneous activity and responded to carbachol and high-K(+) solutions. CONCLUSION: These results show that in fetuses with BOO and dilated ureters absolute ureteric contractile activity is diminished. However, there is functional reorganization of the muscle layers that generates more force in the longitudinal rather than the transverse axis. This would contribute to a reduced ability of the ureter to propel urine and contribute to the development of raised upper tract pressures. PMID- 18410439 TI - Evaluation of dynamic sentinel lymph node biopsy in patients with squamous cell carcinoma of the penis and palpable inguinal nodes. AB - OBJECTIVE: To evaluate dynamic sentinel lymph node biopsy (DSLNB) in patients with squamous cell carcinoma (SCC) of the penis and palpable inguinal lymph nodes, using inguinal lymph node dissection (ILND) as the reference standard to assess the reliability of DSLNB, as using radioscintigraphy and colloidal blue dye injection to locate the SLN was reported to be a useful technique to avoid ILND in men with SCC of the penis and clinically impalpable nodes. PATIENTS AND METHODS: The study included 23 consecutive men with SCC of the penis and clinically palpable inguinal nodes treated between August 1999 and July 2006. On the day before surgery the patient had the SLN located by subcutaneous injection of 60 MBq (99m)Tc-nanocolloid 2 cm proximal to the penile tumour. The following day the patient was taken to the operating room for DSLNB, resection of the penile tumour and simultaneous ILND, if considered indicated (G2-3 and/or T3-4 primary tumour). During surgery 2 mL of colloidal blue dye was injected in the same area as the previous (99m)Tc-nanocolloid injection. The SLNs were located during surgery using a gamma-probe and visualization of blue dye in the node(s), which were then surgically removed. After partial or total penectomy, selected patients had ILND through a 10-cm subinguinal incision. The primary tumour, SLNs and ILND specimens were assessed histopathologically, using haematoxylin and eosin staining only. RESULTS: Biopsy of the primary tumour showed SCC grade 1 in six, grade 2 in 13 and grade 3 in two patients. The clinical T stage was T1 in two, T2 in seven, T3 in 13 and T4 in one. There were clinically palpable inguinal lymph nodes bilaterally in 19 and unilaterally in four men. Scintigraphy before surgery showed inguinal nodes bilaterally in 12 and unilaterally in eight patients, while there were no nodes in three. Surgery comprised partial penectomy in 14, radical penectomy in eight and circumcision alone in one patient. Simultaneous bilateral ILND was done in 15 patients. Inguinal node metastases were present in four of the 23 (17%) patients; the SLN was falsely negative in three (13%), one of whom had a small focus of cancer in the SLN that was missed on initial histopathological examination, and in two the dynamically located SLN contained no cancer, but node metastases were found in the ILND specimen. CONCLUSION: The relatively high false-negative rate of DSLNB indicates that it is not sufficiently reliable to replace complete ILND in men with a high suspicion of nodal metastases, i.e. a high-grade or high-stage primary lesion with clinically palpable inguinal nodes. PMID- 18410440 TI - Application of transcript profiling in formalin-fixed paraffin-embedded diagnostic prostate cancer needle biopsies. AB - OBJECTIVE: To investigate the feasibility of transcript profiling in diagnostic formalin-fixed and paraffin-embedded (FFPE) biopsies for prostate cancer. MATERIALS AND METHODS: Laser-capture microdissection (LCM) was used to microdissect glandular epithelium as well as stromal tissue in archival prostate needle biopsies. Optimized RNA extraction, reverse transcription and real-time PCR (QPCR) protocols were used to detect transcript expression. RNA degradation effects were assessed using hydrolysed cell line RNA and matched xenograft FFPE and frozen tumours. RESULTS: LCM and RNA extraction was achieved in all biopsies from a pilot cohort of five patients. cDNA produced was successfully used to detect expression of glyceraldehyde-3-phosphate dehydrogenase, RPL13, prostate specific antigen, vimentin, inhibitor of differentiation/DNA binding 1 (Id-1) and polycomb group protein enhancer of zeste homolog 2 (EZH2) transcripts. In the cell line and xenograft models, we investigated the effect of RNA degradation on transcript quantification by QPCR. In both models normalization of transcript quantity with a housekeeping gene resulted in restored expression in all degraded samples to within a 50% difference of control samples. Using an extended cohort of 29 biopsies, we tested application in detecting differences in EZH2 and Id-1 expression between malignant and benign epithelium. The results confirmed that our technique was capable of quantifying significant differences in expression between malignant and benign epithelium consistent with the reported trends. CONCLUSION: This study reports the use of standard FFPE needle biopsies for transcript profiling and supports the concept of molecular prognostic studies in tissue acquired at diagnosis in prostate cancer. PMID- 18410441 TI - Mitochondrial DNA in serum of patients with prostate cancer: a predictor of biochemical recurrence after prostatectomy. AB - OBJECTIVE: To investigate the role of circulating mitochondrial DNA (mtDNA) in patients with localized prostate cancer, as recent reports show that patients with advanced cancer have increased levels of mtDNA. PATIENTS AND METHODS: DNA was isolated from the serum of 100 patients with prostate cancer and 18 with benign prostate hyperplasia (BPH). A quantitative real-time polymerase chain reaction was used to amplify 79 bp and 230 bp fragments of the mitochondrial 16s RNA gene, the short fragment representing total mtDNA, including mtDNA truncated by apoptosis, and the long fragment representing mostly mtDNA from other cell death entities. mtDNA integrity was defined as the ratio of long to short mtDNA fragments. RESULTS: The short and long mtDNA levels, and mtDNA integrity, were similar in patients with BPH and cancer (P = 0.940, 0.211 and 0.441, respectively), and were not correlated with clinical or pathological variables, e.g. age, prostate-specific antigen (PSA) level, cT stage, pT stage, seminal vesicle infiltration, lymph node invasion, or Gleason score (P = 0.075 to 0.961). However, patients with high levels of short mtDNA (>75th percentile) had a greater risk of PSA progression and this variable was the strongest predictor of PSA recurrence in a multivariate Cox analysis (P = 0.023; hazard ratio 0.31; 95% confidence interval 0.113-0.851). CONCLUSION: Circulating mtDNA levels did not distinguish between patients with prostate cancer or BPH. However, there was a significant increase in short mtDNA fragments in patients with early PSA recurrence after radical prostatectomy. PMID- 18410442 TI - Histological verification of 11C-choline-positron emission/computed tomography positive lymph nodes in patients with biochemical failure after treatment for localized prostate cancer. AB - OBJECTIVES: To evaluate the potential of (11)C-choline-positron emission tomography (PET)/computed tomography (CT) for planning surgery in patients with prostate cancer and prostate-specific antigen (PSA) relapse after treatment with curative intent. PATIENTS AND METHODS: We retrospectively reviewed the charts of 10 patients with PSA recurrence after either external beam radiation (two) or radical retropubic prostatectomy (eight) for prostate cancer, and who had a laparoscopic lymphadenectomy for suspicious lymph nodes detected on (11)C-choline PET/CT. The histological results and PET/CT findings were compared. RESULTS: In all, 22 suspicious lymph nodes were found on PET/CT, and 14 on conventional CT or magnetic resonance imaging. Comparing the conventional imaging showed concordance in 13 lymph nodes. Three of the 10 patients had no metastatic lymph node disease on definitive histology. The mean (SD) PSA level for these patients was 1.0 (0.4) ng/mL, whereas that in patients with lymph node metastases was 15.1 (9.2) ng/mL (statistically significant difference, P < 0.05). The positive predictive value was seven of 10. All of the patients initially regressed, with PSA increases after lymphadenectomy. Two of the patients are being managed by watchful waiting, two had radiotherapy of the prostate fossa and two had chemotherapy with docetaxel. Four patients were treated by hormone-deprivation therapy. After a mean (SD) follow up of 11 (7) months, one patient died, one has PSA progression, but none of those with negative histology has clinical signs of local recurrence. CONCLUSIONS: (11)C-choline-PET is a valuable tool for detecting recurrent prostate cancer, but the limited positive predictive value should lead to a critical interpretation of the results. PMID- 18410443 TI - Robotic salvage retropubic prostatectomy after radiation/brachytherapy: initial results. AB - OBJECTIVE: To report on the initial four patients who had robotic salvage retropubic prostatectomy (SRP) for biochemical recurrence after radiation therapy, and to review the surgical outcome of robotic cystoprostatectomy for bladder cancer in two patients who previously had prostate cancer. PATIENTS AND METHODS: Since February 2006, four patients had SRP for biochemical failure after radiation and/or brachytherapy. Transrectal biopsy of the prostate confirmed locally recurrent disease and a metastatic evaluation including bone scan and computed tomography of the abdomen and pelvis were negative in all cases. The SRP was done using a six-port transperitoneal approach. An additional two patients had a robotic cystoprostatectomy for bladder cancer, in whom radiation was provided previously for prostate cancer. A retrospective analysis of the immediate and short-term surgical outcome was reviewed. RESULTS: SRP was completed in all patients with no major complication or conversion to an open approach. The mean operative duration was 125 min, the mean (range) blood loss was 117 (50-250) mL and the mean hospital stay was 2.7 days. Of the four patients undergoing SRP, three had extracapsular extension and the first two had positive margins, while no patients had rectal injuries or significant blood loss. The lymph nodes were negative in all the patients. Three patients were continent within a month while one continued to use two to three pads/day at 3 weeks of follow-up. In the two patients who had cystoprostatectomy there were no major complications or increased surgical difficulty. CONCLUSIONS: SRP is technically possible and with limited perioperative morbidity. Further studies are warranted to validate the oncological and functional outcomes of SRP after radiation and/or brachytherapy. Moreover, the robotic approach for radical cystoprostatectomy in patients who have had prostate radiation is feasible, with no increase in perioperative morbidity. PMID- 18410444 TI - Neoadjuvant targeted therapy and advanced kidney cancer: observations and implications for a new treatment paradigm. AB - OBJECTIVE: To evaluate our early experience with neoadjuvant therapy (sunitinib or sorafenib) in advanced renal cell carcinoma (RCC), to explore the effect on both tumour biology and potential for downstaging advanced tumours, as systemic therapy for RCC has historically resulted in little if any primary tumour response, but recent experience with targeted therapy suggests otherwise. PATIENTS AND METHODS: The preliminary experience with neoadjuvant therapy for the surgical management of RCC was reviewed at two large referral centres. Several unique patients were identified who had a novel response to systemic therapy that altered the surgical strategy. RESULTS: Four patients who had targeted therapy before surgery are described and in whom there were effects on tumour biology not seen previously with chemotherapy and cytokine therapy. The selected patients who had neoadjuvant targeted therapy had shrinkage of a tumour thrombus in the inferior vena cava, nodal involvement, renal fossa recurrence and tumour within a solitary kidney. CONCLUSIONS: The introduction of new molecular agents has revolutionized the treatment of patients with metastatic RCC. Responses to targeted therapy within the primary tumour, tumour thrombus, renal fossa recurrence, and lymph node metastases are novel findings not seen during treatment with immunotherapeutic-based strategies. This might be a signal for urological surgeons to re-evaluate the paradigm for the surgical management of advanced RCC. Potential applications are presented to encourage further investigations with targeted therapy in the neoadjuvant setting. PMID- 18410445 TI - The opiorphin gene (ProL1) and its homologues function in erectile physiology. AB - OBJECTIVE: To determine if ProL1, a member of the opiorphin family of genes, can modulate erectile physiology, as it encodes a peptide which acts as a neutral endopeptidase inhibitor, other examples of which (Vcsa1, hSMR3A) modulate erectile physiology. MATERIALS AND METHODS: We cloned members of the opiorphin family of genes into the same mammalian expression backbone (pVAX); 100 microg of these plasmids (pVAX-Vcsa1, -hSMR3A, -hSMR3B and -ProL1) were injected intracorporally into retired breeder rats and the affect on erectile physiology assessed visually, by histology and by measuring the intracavernous pressure (ICP) and blood pressure (BP). As a positive control, rats were treated with pVAX hSlo (expressing the MaxiK potassium channel) and as a negative control the empty backbone plasmid was injected (pVAX). We also compared the level of expression of ProL1 in corporal tissue of patients not reporting erectile dysfunction (ED), ED associated with diabetes and ED not caused by diabetes. RESULTS: Gene transfer of plasmids expressing all members of the opiorphin family had a similar and significant effect on erectile physiology. At the concentration used in these experiments (100 microg) they resulted in higher resting ICP, and histological and visual analysis showed evidence of a priapic-like condition. After electrostimulation of the cavernous nerve, rats had significantly better ICP/BP than the negative control (pVAX). Gene transfer of pVAX-hSlo increased the ICP/BP ratio to a similar extent to the opiorphin homologues, but with no evidence for a priapic-like condition. Corpora cavernosa tissue samples obtained from men with ED, regardless of underlying causes, had significant down-regulation of both hSMR3A and ProL1. CONCLUSION: All members of the human opiorphin family of genes can potentially modulate erectile physiology. Both hSMR3 and ProL1 are down regulated in the corpora of men with ED, and therefore both genes can potentially act as markers of ED. PMID- 18410447 TI - The addition of liposomal doxorubicin to bortezomib, thalidomide and dexamethasone significantly improves clinical outcome of advanced multiple myeloma. AB - Relapsed/refractory myeloma has a poor outcome because of multi-drug resistance, patient low-performance status and toxicity of conventional chemotherapy. To improve results, standard chemotherapeutics and drugs targeting the microenvironment are applied at the same time. Bortezomib, by inhibiting proteasome function, may enhance chemosensitivity to other drugs and overcome drug-resistance. Notably, doxorubicin and bortezomib may reciprocally increase their efficacy. Thus, to improve outcome whilst minimizing therapy-related toxicity, liposomal doxorubicin was added to a bortezomib-based combination. From January 2004, relapsed/refractory myeloma patients referred to our Institution received bortezomib 1.0 mg/m(2) i.v. twice weekly for 2 weeks in a 28-d cycle for up to six cycles, oral dexamethasone 24 mg with the standard scheduling and thalidomide 100 mg continuously (VTD). From January 2005, liposomal doxorubicin, 50 mg/m(2) (30 mg/m(2) for patients older than 75 years), was added on day 4 of each cycle [VTD plus Myocet (MyVTD)]. In total, 70 patients were treated: 28 received VTD and 42 MyVTD. Baseline demographic and clinical characteristics were similar between the two groups. Toxicity was manageable although more pronounced with MyVTD. The overall response rate (81% vs. 50%, P = 0.009), time to progression (19 vs. 11 months, P = 0.01) and progression-free survival (15 vs. 8 months, P = 0.001) were significantly higher with MyVTD regimen, suggesting an improved quality of response. PMID- 18410448 TI - Inferring relative numbers of human leucocyte genome replications. AB - Genome duplication inevitably results in replication errors. A priori, the more times a genome is copied, the greater the average number of replication errors. This principle could be used to 'count' mitotic divisions. Although somatic mutations are rare, cytosine methylation is also copied after DNA replication, but measurably increases with aging at certain CpG rich sequences in mitotic tissues, such as the colon. To further test whether such age-related methylation represents replication errors, these CpG rich 'clock' sequences were measured in leucocytes. Leucocytes within an individual have identical chronological ages (time since birth) but their mitotic ages (numbers of divisions since the zygote) may differ. Neutrophils, B-lymphocytes, and red cell progenitors are produced from relatively quiescent stem cells throughout life, but T-lymphocyte production largely ceases after puberty when the thymus disappears. However, T-lymphocyte genomes may continue to replicate throughout life in response to immunological stimulation. Consistent with this biology, clock methylation significantly increased with aging for T-lymphocyte genomes, but no significant increase was measured in other cell populations. Moreover, this methylation was greater in genomes isolated from their corresponding neoplastic populations. These studies tentatively support the hypothesis that methylation at certain CpG rich sequences in leucocytes could record their mitotic ages. PMID- 18410449 TI - Venous thromboembolism in hospitalised patients: a public health crisis? AB - The majority of hospitalised patients are at risk of venous thrombosis and one in ten hospital deaths are due to pulmonary embolism. A report from the House of Commons Health Committee published in 2005 stated that each year over 25 000 people in England die from venous thromboembolism developing during hospitalisation. The report also stated that, whilst many of these deaths are preventable by thromboprophylaxis, this is not as widely used as it should be. This article reflects on the background prompting the report of the Health Committee and the subsequent response to this, including the publication of a National Institute for Health and Clinical Excellence guideline on reducing the risk of venous thromboembolism in patients undergoing surgery and the strategy for venous thrombosis prevention outlined by the Chief Medical Officer for England. It is anticipated that, by 2009, risk assessment will be mandatory for all hospitalised patients in England; the same strategy is likely for the whole UK. PMID- 18410446 TI - Fatty acid synthase is a novel therapeutic target in multiple myeloma. AB - This study investigated the biological significance of the inhibition of fatty acid synthase (FAS) in multiple myeloma (MM) using the small molecule inhibitor Cerulenin. Cerulenin triggered growth inhibition in both MM cell lines and MM patient cells, and overcame the survival and growth advantages conferred by interleukin-6, insulin-like growth factor-1, and bone marrow stromal cells. It induced apoptosis in MM cell lines with only modest activation of caspase -8, -9, -3 and PARP; moreover, the pan-caspase inhibitor Z-VAD-FMK did not inhibit Cerulenin-induced apoptosis and cell death. In addition, treatment of MM cells with Cerulenin primarily up-regulated apoptosis-inducing factor/endonuclease G, mediators of caspase-independent apoptosis. Importantly, Cerulenin induced endoplasmic reticulum stress response via up-regulation of the Grp78/IRE1alpha/JNK pathway. Although the C-Jun-NH(2)-terminal kinase (JNK) inhibitor SP600215 blocked Cerulenin-induced cytotoxicity, it did not inhibit apoptosis and caspase cleavage. Furthermore, Cerulenin showed synergistic cytotoxic effects with various agents including Bortezomib, Melphalan and Doxorubicin. Our results therefore indicate that inhibition of FAS by Cerulenin primarily triggered caspase-independent apoptosis and JNK-dependent cytotoxicity in MM cells. This report demonstrated that inhibition of FAS has anti-tumour activity against MM cells, suggesting that it represents a novel therapeutic target in MM. PMID- 18410450 TI - Relapse prediction in acute myeloid leukaemia patients in complete remission using WT1 as a molecular marker: development of a mathematical model to predict time from molecular to clinical relapse and define optimal sampling intervals. AB - We hypothesized that Wilms tumour 1 gene (WT1) expression levels in acute myeloid leukaemia (AML) patients might have predictive value and reveal molecular relapse kinetics. WT1 level was determined at diagnosis, during therapy and post-therapy follow-up in 89 patients who reached first complete remission (CR1) (952 samples, median 8 samples/patient, range 2-38). CR1 bone marrow (BM) WT1 level above normal (based on 39 healthy donors) was an independent adverse prognostic factor regarding both disease-free survival [hazard ratio (HR) 4.46, P = 0.001] and overall survival (HR 2.62, P = 0.019). By grouping 34 BM and 99 peripheral blood (PB) complete remission samples in monthly intervals prior to clinical relapse, disease development was delineated and a simple mathematical model constructed, that allowed for the prediction of relapse detection rates (RDRs) as well as median times [t(m)s] from WT1 positivity to clinical relapse. BM sampling was required to obtain RDRs above 93% and t(m)s above 67 d. Acceptable RDRs and t(m)s (81% and 44 d, respectively) could be acquired by bimonthly PB sampling. In conclusion, CR1 WT1 expression is an independent prognostic factor in AML. According to our model, BM is superior for relapse prediction, but PB samples are useful when shorter sampling intervals are possible. PMID- 18410451 TI - JAK2 V617F mutation is associated with increased risk of thrombosis in Chinese patients with essential thrombocythaemia. PMID- 18410452 TI - Expression of P2X5 in lymphoid malignancies results in LRH-1-specific cytotoxic T cell-mediated lysis. AB - Minor histocompatibility antigens (MiHA) selectively expressed by haematopoietic cells are attractive targets for specific immunotherapy after allogeneic stem cell transplantation (SCT). Previously, we described LRH-1 as a haematopoietic lineage restricted MiHA that is capable of eliciting an allogeneic cytotoxic T lymphocyte (CTL) response after SCT and donor lymphocyte infusion. Importantly, the gene encoding LRH-1, P2X5, is not expressed in prominent graft-versus-host disease target tissues such as skin, liver and gut. Here, we investigate whether LRH-1-specific immunotherapy may be exploited for the treatment of lymphoid malignancies. We examined P2X5 mRNA expression in a large panel of patient samples and cell lines from different types of lymphoid malignancies by real-time quantitative reverse transcription polymerase chain reaction. P2X5 mRNA was highly expressed in malignant cells from all stages of lymphoid development. Furthermore, all LRH-1-positive lymphoid tumour cell lines were susceptible to LRH-1 CTL-mediated lysis in flow cytometry-based cytotoxicity assays. However, interferon-gamma production was low or absent after stimulation with some cell lines, possibly due to differences in activation thresholds for CTL effector functions. Importantly, primary cells from patients with lymphoid malignancies were effectively lysed by LRH-1-specific CTL. These findings indicate that MiHA LRH-1 is a potential therapeutic target for cellular immunotherapy of lymphoid malignancies. PMID- 18410453 TI - Advances in the understanding of mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is a well-defined lymphoid neoplasm characterized by a proliferation of mature B lymphocytes expressing CD5 that may show a spectrum of morphological and phenotypic features broader than initially described. Although some patients may follow an indolent clinical evolution, in most of them the tumour has an aggressive behaviour with poor response to conventional chemotherapy. The genetic hallmark is the t(11;14)(q13;q32) translocation leading to the overexpression of cyclin D1, which is considered the initial oncogenic event. In addition to this translocation, MCL may carry a high number of secondary chromosomal and molecular alterations that target regulatory elements of the cell cycle machinery and senescence (BMI1/INK4/ARF/CDK4/RB1), DNA damage response pathways (ATM/CHK2/p53), and cell survival signals. The knowledge of these mechanisms and their influence on the behaviour of the tumour are facilitating the development of prognostic models with a more precise prediction of the clinical evolution of the patients. This information coupled with the availability of a new generation of innovative drugs targeting basic molecular process of the tumour cells, should facilitate the design of new therapeutic protocols able to overcome the resistance of this aggressive lymphoma to conventional treatments and improve the life expectancy of the patients. PMID- 18410454 TI - Prediction of survival using absolute lymphocyte count for newly diagnosed patients with multiple myeloma: a retrospective study. AB - Absolute lymphocyte count (ALC) recovery after autologous stem cell transplantation for multiple myeloma (MM) has been reported to be an independent prognostic factor for clinical outcome. The role of ALC on survival in newly diagnosed untreated MM patients is unknown. Between 1994 and 2002, we analysed retrospectively 537 MM patients of 1835 consecutive MM patients that were neither uniformly treated nor part of a clinical trail, but originally diagnosed and followed at the Mayo Clinic. The primary endpoint was to assess the role of ALC at the time of MM diagnosis on overall survival (OS). The median follow-up was 35.1 months (range: 1-152.5 months). ALC, as a continuous variable, was identified as prognostic factor for OS (Hazard ratio = 0.473, 95% confidence interval = 0.359-0.618, P < 0.0001). MM patients with an ALC >/=1.4 x 10(9)/l experienced superior OS compared with MM patients with an ALC <1.4 x 10(9)/l (65 vs. 26 months, P < 0.0001). Multivariate analysis identified ALC as an independent prognostic factor for OS. This study showed that, in newly diagnosed MM, ALC is an independent prognostic factor for OS, suggesting a significant role of host immune status in the survival of MM. PMID- 18410455 TI - The use of intravenous iron in patients with cancer-related anaemia. AB - Intravenous iron has become the standard of care in patients with renal failure receiving treatment with erythropoiesis stimulating agents (ESAs) to treat true and functional iron deficiency and to prevent its development in haemodialysis patients. In cancer-related anaemia, several recently published, randomised studies suggested that intravenous iron improved haemoglobin response rates in ESA-treated patients compared to those treated with oral iron or placebo. The data supporting the efficacy of intravenous iron instead of oral iron in this setting are increasingly persuasive but larger randomised trials are needed before definitive recommendations are made. PMID- 18410456 TI - Chronic lymphocytic leukaemia and acute myeloid leukaemia are not associated with AKT1 pleckstrin homology domain (E17K) mutations. PMID- 18410457 TI - Farnesyltransferase inhibitors and their potential role in therapy for myelodysplastic syndromes and acute myeloid leukaemia. AB - Novel strategies are required for treatment of acute myeloid leukaemia (AML) and higher risk myelodysplastic syndrome (MDS) patients who are not eligible for intensive chemotherapy and/or allogenic stem cell transplantation. As activating RAS mutations are frequent in these diseases, one novel approach, consisting of interfering with isoprenylation of RAS proteins by farnesyltransferase inhibitors (FTIs), has been proposed. Clinical phase II studies with the oral FTIs tipifarnib and lonafarnib in previously untreated AML, MDS and chronic myelomonocytic leukaemia yielded rather encouraging results while results in relapsed and/or refractory AML were disappointing. Results of a phase III trial in untreated AML in the elderly with tipifarnib were also disappointing. Clinical responses were not related to RAS mutations, suggesting additional actions of FTIs on other molecular targets. The combination of existing FTIs with other treatments, such as chemotherapy (in AML) and hypomethylating agents (in MDS and AML), is under investigation. Ongoing studies will also determine if gene profiling analysis may help to identify patients that will respond to FTIs. PMID- 18410458 TI - Feasibility of prophylaxis and immune tolerance induction regimens in haemophilic children using fully implantable central venous catheters. AB - Venous access represents the major barrier to the feasibility of prophylaxis and immune tolerance induction (ITI) in haemophilic children. Ports improve treatment feasibility, but their duration is limited by infectious complications. This study aimed at evaluating whether or not ports allow haemophilic children to maintain the treatment regimen in the long term. Children were prospectively followed-up and underwent port removal either for complications or transition to peripheral veins. Of 27 ports (17 used for prophylaxis and 10 for ITI), 25 were removed after a median of 3.3 years. Inhibitor children showed a younger age at port insertion (P = 0.02), an earlier occurrence of infections (P = 0.006) at a higher rate (P = 0.00001) and an earlier removal for infection (P = 0.05) than non-inhibitor patients. Daily port use was associated with earlier infections at a higher rate compared to less frequent use (P = 0.02). Port removal after a median of 0.8 years prevented ITI completion in 50% of children, while it hampered the maintenance of prophylaxis in 27% of patients. This study showed that ports improved the feasibility of prophylaxis in the majority of non inhibitor children, while they were not suitable for inhibitor children who require a prolonged ITI regimen with daily infusions. PMID- 18410459 TI - Reassessing insurers' access to genetic information: genetic privacy, ignorance, and injustice. AB - Many countries have imposed strict regulations on the genetic information to which insurers have access. Commentators have warned against the emerging body of legislation for different reasons. This paper demonstrates that, when confronted with the argument that genetic information should be available to insurers for health insurance underwriting purposes, one should avoid appeals to rights of genetic privacy and genetic ignorance. The principle of equality of opportunity may nevertheless warrant restrictions. A choice-based account of this principle implies that it is unfair to hold people responsible for the consequences of the genetic lottery, since we have no choice in selecting our genotype or the expression of it. However appealing, this view does not take us all the way to an adequate justification of inaccessibility of genetic information. A contractarian account, suggesting that health is a condition of opportunity and that healthcare is an essential good, seems more promising. I conclude that if or when predictive medical tests (such as genetic tests) are developed with significant actuarial value, individuals have less reason to accept as fair institutions that limit access to healthcare on the grounds of risk status. Given the assumption that a division of risk pools in accordance with a rough estimate of people's level of (genetic) risk will occur, fairness and justice favour universal health insurance based on solidarity. PMID- 18410460 TI - Administrative gatekeeping - a third way between unrestricted patient advocacy and bedside rationing. AB - The inevitable need for rationing of healthcare has apparently presented the medical profession with the dilemma of choosing the lesser of two evils. Physicians appear to be obliged to adopt either an implausible version of traditional professional ethics or an equally problematic ethics of bedside rationing. The former requires unrestricted advocacy of patients but prompts distrust, moral hazard and unfairness. The latter commits physicians to rationing at the bedside; but it is bound to introduce unfair inequalities among patients and lack of political accountability towards citizens. In this paper I shall argue that this dilemma is false, since a third intermediate alternative exists. This alternative, which I term 'administrative gatekeeping', makes it possible for physicians to be involved in rationing while at the same time being genuine advocates of their patients. According to this ideal, physicians are required to follow fair rules of rationing adopted at higher organizational levels within healthcare systems. At the same time, however, they are prohibited from including considerations of cost in their clinical decisions. PMID- 18410461 TI - The medical decision-making process and the family: the case of breast cancer patients and their husbands. AB - OBJECTIVES: The objectives of the study were (1) to assess similarities and differences between breast cancer patients and their husbands in terms of doctor patient/spouse relationships and shared decision making; and (2) to investigate the association between breast cancer patients and husbands in terms of preference of type of doctor, doctor-patient relationship, and shared decision making regarding medical treatment. METHOD: Fifty-seven women with breast cancer, and their husbands, completed questionnaires measuring doctor-patient/spouse relationships (paternalism, autonomy), and decision making regarding medical treatment. RESULTS: Patients believe they have a key role in the medical decision making process (93%) and that the participation of their husbands, and their agreement with the decision, is important (84% and 89%, respectively). Both breast cancer patients and their husbands prefer a shared decision-making process to paternalistic or autonomy-based approaches. CONCLUSION: In contrast to legal and bioethical approaches, which focus on the patient as the primary decision maker, this study reflects a practical recognition of the role of the breast cancer patient's husband in the decision-making process. It also reflects a relational rather than an individualistic perception of patient autonomy. PMID- 18410462 TI - Beyond abortion: the looming battle over death in the 'culture wars'. AB - By concentrating on abortion, the culture wars have avoided facing a crisis about the end of life. This paper explores four themes: (1) the technological transformation of birth and death into matters of decision, not matters of fact; (2) abortion as the nexus of Eros (sex) with Thanatos (death); (3) the real crisis, conveniently masked by our obsession with sex, looming at the end of life, not at its beginning; (4) the surplus-repression that protects us from assuming responsibility for choosing between life and death. PMID- 18410463 TI - Bioethics in China. AB - Historically, the preconditions for the emergence of bioethics in China. were political reforms and their applications. The Hanzhong Euthanasia Case and the publication of Qiu Ren-zong's academic work Bioethics played a significant role in the development of bioethics in China. Other contributory factors include the establishment of the Chinese Society of Medical Ethics/Chinese Medical Association (C.M.A), the publication of the Journal of Chinese Medical Ethics, and the teaching and education of bioethics in China. Major achievements of bioethics in China include the establishment of ethics committee and ethics review system, active international communication and cooperation among the academic circles, and the successful management of the 8th World Congress of Bioethics in Beijing in 2006. Chinese bioethics focus on native Chinese realities and conditions, absorb the international research achievements in relevant fields, and combine international ideas with traditional Chinese doctrines. Admittedly, there are still some aspects to be improved, yet bioethics has attracted a lot of attention from the core leadership in China and has gained sound financial support, which augers well for its further development. This article also briefly introduces the development of bioethics in Hong Kong and Taiwan, China. PMID- 18410464 TI - Brain histamine H1 receptor occupancy of orally administered antihistamines, bepotastine and diphenhydramine, measured by PET with 11C-doxepin. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: 'Bepotastine besilate' is a novel second-generation antihistamine developed in Japan and its antiallergic effects have already been demonstrated by various studies. However, only a few clinical studies regarding its sedative property are available. In addition, histamine H(1) receptor occupancy (H(1)RO) of this new antihistamine has never been measured by positron emission tomography (PET). WHAT THIS STUDY ADDS: This paper provides the first measurement result of cerebral H(1)RO of bepotastine besilate (approximately 15%) as determined by PET. This result is in accordance with the clinical classification of bepotastine as a second-generation antihistamine. In addition, the relationship between subjective sleepiness and cerebral H(1)RO of this second-generation antihistamine is demonstrated for the first time using a placebo-controlled crossover study design. AIMS Antihistamines are frequently used for treating various allergic diseases, but often induce sedation. The degree of sedation can be evaluated by measuring histamine H(1) receptor occupancy (H(1)RO) in the brain using positron emission tomography (PET). The aim was to measure H(1)RO of bepotastine, a new second-generation antihistamine, and to compare it with that of diphenhydramine. METHODS: Eight healthy male volunteers (mean age +/- SD 24.4 +/- 3.3 years) were studied after single oral administration of bepotastine (10 mg), diphenhydramine (30 mg) or placebo, by PET imaging with (11)C-doxepin in a crossover study design. Binding potential ratio and H(1)ROs were calculated using placebo data and were compared between bepotastine and diphenhydramine in the anterior and posterior cingulate gyri (ACG and PCG, respectively), superior and inferior frontal cortices (SFC and IFC, respectively), orbitofrontal cortex (OFC), insular cortex (IC), lateral and medial temporal cortices (LTC and MTC, respectively), parietal cortex (PC), occipital cortex (OC) and sensorimotor cortex (SMC). Plasma concentration of each antihistamine was measured, and its correlation to H(1)RO was examined. RESULTS: H(1)RO after bepotastine treatment was significantly lower than that after diphenhydramine treatment in all cortical regions (P < 0.001). Mean H(1)ROs of bepotastine and diphenhydramine were 14.7% and 56.4%, respectively. H(1)ROs of both bepotastine and diphenhydramine correlated to their respective drug plasma concentration (P < 0.001). CONCLUSION: Oral bepotastine (10 mg), with its relatively low H(1)RO and thus minimal sedation, has the potential for use as a mildly or slightly sedative antihistamine in the treatment of various allergic disorders. PMID- 18410465 TI - Complex regional pain syndrome type I in children. AB - BACKGROUND: Complex Regional Pain Syndrome type I (CRPS I) is a potentially incapacitating syndrome which can occur after a minor injury or operation to a limb. It is a disorder characterized by pain, sensory and motor disturbances. CRPS I is well known in adults, but a relatively new diagnostic entity in children. The clinical presentation of CRPS I in children is, to some extent, different from adults and therefore sometimes not recognized early. The aim of this study was to search for differences in patient characteristics between children and adults with CRPS I. METHODS: We have performed a retrospective chart review of 78 children (age or =15 mm. RESULTS: UTIs developed in 4 of 106 infants from group I and 5 of 19 infants from group II. Surgical interventions were performed on 1 of 106 patients of group I and 7 of 19 patients of group II. These differences were statistically significant (p-values 0.004 and <0.001, respectively). In group I, 6 of 106 patients had VUR; none of them required surgical intervention and only two developed a UTI (one of whom also had contralateral ureteropelvic junction obstruction). Five of 19 infants in group II had underlying VUR, four of them with associated anomalies, 1 infant required surgical correction and 4 developed UTIs. CONCLUSION: Infants with ANH up to 15 mm have a low incidence of UTIs and surgery and a low incidence and benign course of underlying VUR. Therefore, noninvasive postnatal follow-up is justified and standard voiding cystourethrography should not be performed, but only in cases of ureteric dilatation. PMID- 18410469 TI - Prevention of spontaneous abortion in the CBA x DBA/2 mouse model by intravaginal TGF-beta and local recruitment of CD4+8+ FOXP3+ cells. AB - PROBLEM: Activation of latent transforming growth factor (TGF)-beta in seminal plasma has been suggested by Robertson et al. to promote maternal tolerance to paternal antigens. A possible consequence reported by Tremellen et al. is increased pregnancy rates in women undergoing IVF. A decreased spontaneous abortion rate has also been postulated. Seminal plasma contains many factors besides TGF-beta, and a critical test of the hypothesis was required. The purpose of the present study was to directly test the effect of pure TGF-beta. METHOD OF STUDY: Pharmaceutical grade bioactive TGF-beta3 with a bovine serum albumin (BSA) carrier 0.1-1% in phosphate-buffered saline (PBS) was given into the vaginal tract of CBA/J female mice at the time of mating with DBA/2 males. One microgram Salmonella enteritidis lipopolysaccharide was given intraperitoneally to augment occult losses and spontaneous resorptions assessed on day 13.5 of pregnancy. The effect of TGF-beta3 on recruitment of lymphomyeloid cells to the vaginal wall and vaginal lumen of unmated mice in estrus was assessed using immunohistochemistry and flow cytometry. RESULTS: Two nanogram of intravaginal TGF-beta3 in 0.1% BSA PBS or 20 ng in 1% BSA-PBS reduced abortion rates. Protection was comparable to that achieved by immunization with BALB/c spleen cells. Fraction V BSA, a binder of TGF-betas, had some activity, and could reduce availability of added TGF beta3. CD11c dendritic cells, CD3+ T cells, and CD25+ cells were recruited to the vaginal wall by 48 hr after TGF-beta3 treatment, and cellularity of vaginal exudates increased. Foxp3+ cells were present in increased numbers, and appeared to be CD8+ and CD4+ 8+. Semen, but not TGF-beta3, stimulated a physiological polymorphonuclear leukocyte exudate. CONCLUSION: Intravaginal bioactive TGF-beta3 can enhance success of pregnancy in vivo in an established model of abortion. The result could be explained by the independent ability of TGF-beta to promote a regulatory T-cell response. PMID- 18410470 TI - Presence of antisperm antibodies reactive with peptide epitopes of FA-1 and YLP12 in sera of immunoinfertile women. AB - PROBLEM: Recent studies in several laboratories are focused on delineating sperm antigens that are relevant to fertility and examining involvement of antibodies to these antigens in human immunoinfertility. Our laboratory has characterized two such antigens, namely fertilization antigen (FA-1) and YLP12 dodecamer sequence that are involved in sperm-oocyte binding. The present study was conducted to examine the occurrence of isoantibodies to various peptide epitopes of human and murine FA-1 antigen and YLP12 peptide in sera of immunoinfertile and fertile women. METHOD OF STUDY: Sera from 67 immunoinfertile and 19 fertile women were collected. Various peptides based up on human and murine FA-1 antigen and YLP(12) were synthesized, and examined for immunoreactivity with these sera by using ELISA. Four immunodominant sequences, two each from human (hFA-1 82-97aa and hFA-1 200-219aa) and mouse (mFA-1 2-19aa and mFA-1 117-136aa) FA-1 antigen, were selected for the present study. Another human FA-1 sequence, hFA-1 220 240aa, that was not in the immunodominant region was used as a control. RESULTS: For human FA-1 peptides, 41.8% of the immunoinfertile sera reacted positively (>or=2 SD units) with hFA-1 82-97aa, 24.6% (16/65) with hFA-1 200-219aa, and 3% (2/66) with hFA-1 220-240aa peptide. For two murine FA-1 peptides, 41.7% (25/60) of the immunoinfertile sera reacted positively with mFA-1 2-19aa, and 41.5% (27/65) with mFA-1 117-136aa peptide. For the YLP12 dodecamer peptide, 43.3% (29/67) of the immunoinfertile sera reacted positively. None of the sera from fertile women reacted positively with any of these peptides. CONCLUSION: In conclusion, our data indicate that the immunoinfertile women have circulating isoantibodies against at least two immunodominant peptide epitopes of human and murine FA-1 antigen and YLP12 peptide sequence. These peptides may find clinical application in the specific diagnosis and treatment of female infertility and contraceptive vaccine development. PMID- 18410471 TI - Fluvastatin inhibits hepatitis C replication in humans. AB - BACKGROUND Hepatitis C viral (HCV) infection is the leading cause of death due to liver disease in the United States. Currently, pegylated interferon and ribavirin produce sustained viral remission in only 50% of patients. Additional agents are needed to increase the cure rate. In vitro experiments show strong antiviral effects of fluvastatin against HCV. OBJECTIVES: To assess the safety and antiviral effects of fluvastatin in chronic HCV carriers. METHODS: 31 veterans with chronic HCV were prospectively given oral doses of fluvastatin, 20 to 320 mg/day, for 2-12 weeks with weekly monitoring of HCV RNA and liver tests. Reductions of viral load (P < 0.01) versus a control group were considered suppressive. RESULTS: With 80 mg a day or less, 11/22 (50%) patients responded by lowering HCV RNA. The first lowering occurred within 4 weeks (9/11, 82%). The greatest weekly change in HCV RNA level was a 1.75 log(10) reduction. When lowered in responders, the viral load remained relatively constant for 2-5 weeks (7/9, 78%), or on the next test rebounded immediately to a non-significant change from, baseline (n = 2). Continued lowering of virus was seen in 2/19 (22 %) patients when the study ended. We found no evidence of liver tests worsening. CONCLUSIONS: FLV used as monotherapy in vivo showed suppressive effects of HCV clinically that are modest, variable, and often short-lived. These findings support "proof-of-concept" for pilot trials combining fluvastatin with standard therapy. Statins and fluvastatin, in particular, appear to be safe for use in hepatitis C. PMID- 18410472 TI - The power and robustness of maximum LOD score statistics. AB - The maximum LOD score statistic is extremely powerful for gene mapping when calculated using the correct genetic parameter value. When the mode of genetic transmission is unknown, the maximum of the LOD scores obtained using several genetic parameter values is reported. This latter statistic requires higher critical value than the maximum LOD score statistic calculated from a single genetic parameter value. In this paper, we compare the power of maximum LOD scores based on three fixed sets of genetic parameter values with the power of the LOD score obtained after maximizing over the entire range of genetic parameter values. We simulate family data under nine generating models. For generating models with non-zero phenocopy rates, LOD scores maximized over the entire range of genetic parameters yielded greater power than maximum LOD scores for fixed sets of parameter values with zero phenocopy rates. No maximum LOD score was consistently more powerful than the others for generating models with a zero phenocopy rate. The power loss of the LOD score maximized over the entire range of genetic parameters, relative to the maximum LOD score calculated using the correct genetic parameter value, appeared to be robust to the generating models. PMID- 18410473 TI - Alleles of DLA-DRB1 are not unique in German Shepherd dogs having degenerative myelopathy. PMID- 18410474 TI - Genetic variation in the pro-melanin-concentrating hormone gene affects carcass traits in Bos taurus cattle. AB - Mammalian pro-melanin-concentrating hormone (PMCH) has previously been shown to affect feed intake in rodent species. The objectives of this study were to sequence the Bos taurus PMCH gene in order to identify any existing genetic variants and to evaluate whether these affected carcass traits. An A-to-T SNP was identified at position -134 relative to the ATG start codon (g.-134A>T). The alleles at this SNP were significantly associated with average fat and grade fat in two crossbred populations of Bos taurus cattle. The g.-134T allele may introduce a binding site for the transcriptional repressor, adenovirus E4 promoter binding protein, which may contribute to this effect. The g.-134A allele occurred in 67% of cattle examined and was associated with higher fat levels. PMID- 18410475 TI - Assignment of 115 genes from HSA9 and HSA14 to SSC1q by RH mapping to generate a dense human-pig comparative map. AB - A large number of significant QTL for economically important traits including average daily gain have been located on SSC1q, which, as shown by chromosome painting, corresponds to four human chromosomes (HSA9, 14, 15 and 18). To provide a comprehensive comparative map for efficient selection of candidate genes, 81 and 34 genes localized on HSA9 and HSA14 respectively were mapped to SSC1q using a porcine 7000-rad radiation hybrid panel (IMpRH). This study, together with the cytogenetic map (http://www2.toulouse.inra.fr/lgc/pig/cyto/genmar/htm/1GM.HTM), demonstrates that SSC1q2.1-q2.13 corresponds to the region ranging from 44.6 to 63.2 Mb on HSA14q21.1-q23.1, the region from 86.5 to 86.8 Mb on HSA15q24-q25, the region from 0.9 to 27.2 Mb on HSA9p24.3-p21, the region from 35.1 to 38.0 Mb on HSA9p13, the region from 70.3 to 79.3 Mb on HSA9q13-q21 and the region from 96.4 to 140.0 Mb on HSA9q22.3-q34. The conserved synteny between HSA9 and SSC1q is interrupted by at least six sites, and the synteny between HSA14 and SSC1q is interrupted by at least one site. PMID- 18410476 TI - An equine chromosome 3 inversion is associated with the tobiano spotting pattern in German horse breeds. AB - The tobiano white-spotting pattern is one of several known depigmentation phenotypes in horses and is desired by many horse breeders and owners. The tobiano spotting phenotype is inherited as an autosomal dominant trait. Horses that are heterozygous or homozygous for the tobiano allele (To) are phenotypically indistinguishable. A SNP associated with To had previously been identified in intron 13 of the equine KIT gene and was used for an indirect gene test. The test was useful in several horse breeds. However, genotyping this sequence variant in the Lewitzer horse breed revealed that 14% of horses with the tobiano pattern did not show the polymorphism in intron 13 and consequently the test was not useful to identify putative homozygotes for To within this breed. Speculations were raised that an independent mutation might cause the tobiano spotting pattern in this breed. Recently, the putative causative mutation for To was described as a large chromosomal inversion on equine chromosome 3. One of the inversion breakpoints is approximately 70 kb downstream of the KIT gene and probably disrupts a regulatory element of the KIT gene. We obtained genotypes for the intron 13 SNP and the chromosomal inversion for 204 tobiano spotted horses and 24 control animals of several breeds. The genotyping data confirmed that the chromosomal inversion was perfectly associated with the To allele in all investigated horses. Therefore, the new test is suitable to discriminate heterozygous To/+ and homozygous To/To horses in the investigated breeds. PMID- 18410477 TI - Familiality of major depressive disorder and gender differences in comorbidity. AB - OBJECTIVE: Gender differences exist in the prevalence and psychiatric comorbidity of major depressive disorder (MDD). This study investigates whether familiality of MDD contributes to observed gender differences in comorbidity. METHOD: Familial (f-MDD) and non-familial (nf-MDD) MDD cases from a population sample were assessed for comorbid dysthymia, anxiety disorders and alcohol-related disorders using the Composite International Diagnostic Interview (CIDI). Logistic regression analyses were performed to examine the effect of f-MDD on gender differences in comorbidity, adjusted for confounders. RESULTS: Women with f-MDD reported significantly more comorbid dysthymia and generalized anxiety disorder (GAD) than their male counterparts; women with nf-MDD reported significantly more comorbid simple phobias and agoraphobia than their male counterparts. Gender differences in comorbid panic disorder and alcohol-related disorders occurred independently of the familial load. Adjustment for age of onset, severity and recurrence of MDD did not change these results. CONCLUSION: Models to explain comorbidity patterns of MDD differ by gender. Familiality of MDD should be taken into account. PMID- 18410478 TI - Meristem activity during flower and ovule development in tomato is controlled by the mini zinc finger gene INHIBITOR OF MERISTEM ACTIVITY. AB - Plants have the ability to form new organs as a result of indeterminate growth ensured by specific regions of pluripotent cells, called meristems. Flowers are produced by the activity of floral meristems which differ from vegetative meristems in their determinate fate. Transcriptional complexes associating C-, D- and E-type MADS box proteins are responsible for flower determinacy by controlling the stem cell population within the floral meristem. We report here that the INHIBITOR OF MERISTEM ACTIVITY (IMA) gene encoding a mini zinc finger (MIF) protein from tomato (Solanum lycopersicum) regulates the processes of flower and ovule development. IMA inhibits cell proliferation during floral termination, controls the number of carpels during floral development and acts as a repressor of the meristem organizing centre gene WUSCHEL. Underexpression of IMA prevents the formation of gametophytic tissue, which is replaced by an undetermined sporophytic tissue. Molecular analyses of MADS box gene expression revealed that IMA participates in the termination of floral meristems and initiation of ovule primordia by activating D-type gene expression. Taken together, our data indicate that both termination of floral meristems and differentiation of nucelli during ovule development require a similar mechanism involving the repression of WUSCHEL and the activation of D-class genes. PMID- 18410479 TI - A gene expression atlas of the model legume Medicago truncatula. AB - Legumes played central roles in the development of agriculture and civilization, and today account for approximately one-third of the world's primary crop production. Unfortunately, most cultivated legumes are poor model systems for genomic research. Therefore, Medicago truncatula, which has a relatively small diploid genome, has been adopted as a model species for legume genomics. To enhance its value as a model, we have generated a gene expression atlas that provides a global view of gene expression in all major organ systems of this species, with special emphasis on nodule and seed development. The atlas reveals massive differences in gene expression between organs that are accompanied by changes in the expression of key regulatory genes, such as transcription factor genes, which presumably orchestrate genetic reprogramming during development and differentiation. Interestingly, many legume-specific genes are preferentially expressed in nitrogen-fixing nodules, indicating that evolution endowed them with special roles in this unique and important organ. Comparative transcriptome analysis of Medicago versus Arabidopsis revealed significant divergence in developmental expression profiles of orthologous genes, which indicates that phylogenetic analysis alone is insufficient to predict the function of orthologs in different species. The data presented here represent an unparalleled resource for legume functional genomics, which will accelerate discoveries in legume biology. PMID- 18410480 TI - Glycine-rich RNA-binding protein 7 affects abiotic stress responses by regulating stomata opening and closing in Arabidopsis thaliana. AB - Despite the fact that glycine-rich RNA-binding proteins (GRPs) have been implicated in the responses of plants to environmental stresses, their physiological functions and mechanisms of action in stress responses remain largely unknown. Here, we assessed the functional roles of GRP7, one of the eight GRP family members in Arabidopsis thaliana, on seed germination, seedling growth, and stress tolerance under high salinity, drought, or cold stress conditions. The transgenic Arabidopsis plants overexpressing GRP7 under the control of the cauliflower mosaic virus 35S promoter displayed retarded germination and poorer seedling growth compared with the wild-type plants and T-DNA insertional mutant lines under high salinity or dehydration stress conditions. By contrast, GRP7 overexpression conferred freezing tolerance in Arabidopsis plants. GRP7 is expressed abundantly in the guard cells, and has been shown to influence the opening and closing of the stomata, in accordance with the prevailing stress conditions. GRP7 is localized to both the nucleus and the cytoplasm, and is involved in the export of mRNAs from the nucleus to the cytoplasm under cold stress conditions. Collectively, these results provide compelling evidence that GRP7 affects the growth and stress tolerance of Arabidopsis plants under high salt and dehydration stress conditions, and also confers freezing tolerance, particularly via the regulation of stomatal opening and closing in the guard cells. PMID- 18410481 TI - Genome-wide transposon tagging reveals location-dependent effects on transcription and chromatin organization in Arabidopsis. AB - The interphase nucleus exists as a highly dynamic system, the physical properties of which have functional importance in gene regulation. Not only can gene expression be influenced by the local sequence context, but also by the architecture of the nucleus in three-dimensions (3D), and by the interactions between these levels via chromatin modifications. A challenging task is to resolve the complex interplay between sequence- and genome structure-based control mechanisms. Here, we created a collection of 277 Arabidopsis lines that allow the visual tracking of individual loci in living plants while comparing gene expression potential at these locations, via an identical reporter cassette. Our studies revealed regional gene silencing near a heterochromatin island, via DNA methylation, that is correlated with mobility constraint and nucleolar association. We also found an example of nucleolar association that does not correlate with gene suppression, suggesting that distinct mechanisms exist that can mediate interactions between chromatin and the nucleolus. These studies demonstrate the utility of this novel resource in unifying structural and functional studies towards a more comprehensive model of how global chromatin organization may coordinate gene expression over large scales. PMID- 18410482 TI - The cytoplasmic-localized, cytoskeletal-associated RNA binding protein OsTudor SN: evidence for an essential role in storage protein RNA transport and localization. AB - Previous studies have demonstrated that the major storage protein RNAs found in the rice endosperm are transported as particles via actomyosin to specific subdomains of the cortical endoplasmic reticulum. In this study, we examined the potential role of OsTudor-SN, a major cytoskeletal-associated RNA binding protein, in RNA transport and localization. OsTudor-SN molecules occur as high molecular-weight forms, the integrity of which are sensitive to RNase. Immunoprecipitation followed by RT-PCR showed that OsTudor-SN binds prolamine and glutelin RNAs. Immunofluorescence studies using affinity-purified antibodies show that OsTudor-SNs exists as particles in the cytoplasm, and are distributed to both the protein body endoplasmic reticulum (ER) and cisternal ER. Examination of OsTudor-SN particles in transgenic rice plants expressing GFP-tagged prolamine RNA transport particles showed co-localization of OsTudor-SN and GFP, suggesting a role in RNA transport. Consistent with this view, GFP-tagged OsTudor-SN is observed in living endosperm sections as moving particles, a property inhibited by microfilament inhibitors. Downregulation of OsTudor-SN by antisense and RNAi resulted in a decrease in steady state prolamine RNA and protein levels, and a reduction in the number of prolamine protein bodies. Collectively, these results show that OsTudor-SN is a component of the RNA transport particle, and may control storage protein biosynthesis by regulating one or more processes leading to the transport, localization and anchoring of their RNAs to the cortical ER. PMID- 18410483 TI - The Arabidopsis GSQ5/DOG1 Cvi allele is induced by the ABA-mediated sugar signalling pathway, and enhances sugar sensitivity by stimulating ABI4 expression. AB - ABI4 encodes an AP2 family transcription factor that is a central regulator in sugar responsive gene expression in plants. Sugar-induced ABI4 regulates plant genes essential for photosynthesis, and carbon, nitrogen and lipid metabolism. ABI4 activity is induced via the ABA-mediated sugar signalling pathway, which is initiated by the glucose sensing protein hexokinase. Natural variation in sugar sensitivity was used to identify new loci involved in sugar signalling. Five quantitative trait loci (QTLs) for glucose sensitivity (GSQ1-GSQ5) were identified in a Ler/Cvi recombinant inbred line (RIL) population. The GSQ3, GSQ4 and GSQ5 loci are positioned in regions not previously associated with known sugar-sensing genes. GSQ5 was fine mapped and cloned using a candidate-gene approach. The GSQ5 locus was shown to encode the DELAY OF GERMINATION 1 (DOG1) gene. DOG1 was previously identified as a major locus in seed dormancy control. Glucose addition induced the expression of the GSQ5/DOG1 Cvi allele, whereas the Ler and Col alleles did not respond to glucose. Positive feedback was observed between the ABA-mediated sugar signalling pathway and the GSQ5/DOG1 Cvi allele. Expression of the GSQ5/DOG1 Cvi allele requires the ABA-mediated sugar signalling pathway, of which ABI4 is an important component. In addition, sugar induction of ABI4 was promoted by the GSQ5/DOG1 Cvi allele. PMID- 18410484 TI - The MYB98 subcircuit of the synergid gene regulatory network includes genes directly and indirectly regulated by MYB98. AB - The female gametophyte contains two synergid cells that play a role in many steps of the angiosperm reproductive process, including pollen tube guidance. At their micropylar poles, the synergid cells have a thickened and elaborated cell wall: the filiform apparatus that is thought to play a role in the secretion of the pollen tube attractant(s). MYB98 regulates an important subcircuit of the synergid gene regulatory network (GRN) that functions to activate the expression of genes required for pollen tube guidance and filiform apparatus formation. The MYB98 subcircuit comprises at least 83 downstream genes, including 48 genes within four gene families (CRP810, CRP3700, CRP3730 and CRP3740) that encode Cys rich proteins. We show that the 11 CRP3700 genes, which include DD11 and DD18, are regulated by a common cis-element, GTAACNT, and that a multimer of this sequence confers MYB98-dependent synergid expression. The GTAACNT element contains the MYB98-binding site identified in vitro, suggesting that the 11 CRP3700 genes are direct targets of MYB98. We also show that five of the CRP810 genes, which include DD2, lack a functional GTAACNT element, suggesting that they are not directly regulated by MYB98. In addition, we show that the five CRP810 genes are regulated by the cis-element AACGT, and that a multimer of this sequence confers synergid expression. Together, these results suggest that the MYB98 branch of the synergid GRN is multi-tiered and, therefore, contains at least one additional downstream transcription factor. PMID- 18410485 TI - Coexpression of wild-type and mutant prion proteins alters their cellular localization and partitioning into detergent-resistant membranes. AB - Transmissible spongiform encephalopathies (TSEs) are a group of diseases of infectious, sporadic and genetic origin, found in higher organisms and caused by the pathological form of the prion protein. The inheritable subgroup of TSEs is linked to insertional or point mutations in the prion gene prnp, which favour its misfolding and are passed on to offspring in an autosomal-dominant fashion. The large majority of patients with these diseases are heterozygous for the prnp gene, leading to the coexpression of the wild-type (wt) (PrP(C)) and the mutant forms (PrPmut) in the carriers of these mutations. To mimic this situation in vitro, we produced Fischer rat thyroid cells coexpressing PrPwt alongside mutant versions of mouse PrP including A117V, E200K and T182A relevant to the human TSE diseases Gestmann-Straussler-Scheinker (GSS) disease and familial Creutzfeldt Jakob disease (fCJD). We found that coexpression of mutant PrP with wt proteins does not affect the glycosylation pattern or the biochemical characteristics of either protein. However, FRET and co-immunoprecipitation experiments suggest an interaction occurring between the wt and mutant proteins. Furthermore, by comparing the intracellular localization and detergent-resistant membrane (DRM) association in single- and double-expressing clones, we found changes in the intracellular/surface ratio and an increased sequestration of both proteins in DRMs, a site believed to be involved in the pathological conversion (or protection thereof) of the prion protein. We, therefore, propose that the mutant forms alter the subcellular localization and the membrane environment of the wt protein in co-transfected cells. These effects may play a role in the development of these diseases. PMID- 18410486 TI - The ubiquitin E3 ligase MARCH7 is differentially regulated by the deubiquitylating enzymes USP7 and USP9X. AB - Protein modification by one or more ubiquitin chains serves a critical signalling function across a wide range of cellular processes. Specificity within this system is conferred by ubiquitin E3 ligases, which target the substrates. Their activity is balanced by deubiquitylating enzymes (DUBs), which remove ubiquitin from both substrates and ligases. The RING-CH ligases were initially identified as viral immunoevasins involved in the downregulation of immunoreceptors. Their cellular orthologues, the Membrane-Associated RING-CH (MARCH) family represent a subgroup of the classical RING genes. Unlike their viral counterparts, the cellular RING-CH proteins appear highly regulated, and one of these in particular, MARCH7, was of interest because of a potential role in neuronal development and lymphocyte proliferation. Difficulties in detection and expression of this orphan ligase lead us to search for cellular cofactors involved in MARCH7 stability. In this study, we show that MARCH7 readily undergoes autoubiquitylation and associates with two deubiquitylating enzymes - ubiquitin-specific protease (USP)9X in the cytosol and USP7 in the nucleus. Exogenous expression and short interfering RNA depletion experiments demonstrate that MARCH7 can be stabilized by both USP9X and USP7, which deubiquitylate MARCH7 in the cytosol and nucleus, respectively. We therefore demonstrate compartment specific regulation of this E3 ligase through recruitment of site-specific DUBs. PMID- 18410487 TI - AP-1 and AP-3 mediate sorting of melanosomal and lysosomal membrane proteins into distinct post-Golgi trafficking pathways. AB - The adaptor complexes AP-1 and AP-3 are localized to endosomes and/or the trans Golgi network (TGN). Because of limitations in analysing intracellular adaptor function directly, their site of function is a matter of ongoing uncertainty. To overcome this problem and to analyse adaptor sorting at the TGN, we reconstituted vesicle formation from Golgi/TGN-enriched membranes in a novel in vitro budding assay. Melanocytes were metabolically labelled followed by a 19 degrees C temperature block to accumulate newly synthesized proteins in Golgi membranes, which were then enriched by subcellular fractionation and used as donor membranes for vesicle formation in vitro. The incorporation of the melanosomal proteins tyrosinase and tyrosinase-related protein 1 (TRP-1) as well as Lamp-1 and 46 kDa mannose-6-phosphate receptor (MPR46) into Golgi/TGN-derived vesicles was temperature, nucleotide, cytosol, ADP ribosylation factor 1 and adaptor dependent. We show that sorting of TRP-1 and MPR46 was AP-1 dependent, while budding of tyrosinase and Lamp-1 required AP-3. Depletion of clathrin inhibited sorting of all four cargo proteins, suggesting that AP-1 and AP-3 are involved in the formation of distinct types of clathrin-coated vesicles, each of which is characterized by the incorporation of specific cargo membrane proteins. PMID- 18410489 TI - Heat acclimation and cross-tolerance against anoxia in Arabidopsis. AB - Arabidopsis seedlings are highly sensitive to low oxygen and they die rapidly when exposed to anoxia. Tolerance to anoxia depends on the ability to efficiently use carbohydrates through the fermentative pathway, as highlighted by the lower tolerance displayed by a mutant devoid of alcohol dehydrogenase. Other mechanisms of tolerance are also possible and may include a role for heat-induced genes. In fact, heat shock proteins (HSPs) are induced by anoxia. This suggests that there may be a cross-adaptation mechanism between heat and anoxic stress, and in this work, we studied the acclimation of Arabidopsis seedlings both to low oxygen and heat. The results show that seedlings subjected to hypoxia or heat pretreatment survive anoxia much better. Interestingly, we also observed an increased anoxia tolerance in heat-treated alcohol dehydrogenase (adh) mutant plants. On the other hand, anoxic pretreatment does not confer tolerance to heat stress. The success of the induction of HSPs by anoxia is in direct relation to the amount of sucrose available, and this in turn relates to how well seedlings will survive under anoxia. HSP transcripts were also detected during seed development and germination, two hypoxia-prone processes, suggesting that hypoxia-induced HSP expression is physiologically relevant. PMID- 18410488 TI - Arp11 affects dynein-dynactin interaction and is essential for dynein function in Aspergillus nidulans. AB - The dynactin complex contains proteins including p150 that interacts with cytoplasmic dynein and an actin-related protein Arp1 that forms a minifilament. Proteins including Arp11 and p62 locate at the pointed end of the Arp1 filament, but their biochemical functions are unclear (Schroer TA. Dynactin. Annu Rev Cell Dev Biol 2004;20:759-779). In Aspergillus nidulans, loss of Arp11 or p62 causes the same nuclear distribution (nud) defect displayed by dynein mutants, indicating that these pointed-end proteins are essential for dynein function. We constructed a strain with S-tagged p150 of dynactin that allows us to pull down components of the dynactin and dynein complexes. Surprisingly, while the ratio of pulled-down Arp1 to S-p150 in Arp11-depleted cells is clearly lower than that in wild-type cells, the ratio of pulled-down dynein to S-p150 is significantly higher. We further show that the enhanced dynein-dynactin interaction in Arp11 depleted cells is also present in the soluble fraction and therefore is not dependent upon the affinity of these proteins to the membrane. We suggest that loss of the pointed-end proteins alters the Arp1 filament in a way that affects the conformation of p150 required for its proper interaction with the dynein motor. PMID- 18410490 TI - Seasonal differences in photosynthesis between the C3 and C4 subspecies of Alloteropsis semialata are offset by frost and drought. AB - The regional abundance of C(4) grasses is strongly controlled by temperature, however, the role of precipitation is less clear. Progress in elucidating the direct effects of photosynthetic pathway on these climate relationships is hindered by the significant genetic divergence between major C(3) and C(4) grass lineages. We addressed this problem by examining seasonal climate responses of photosynthesis in Alloteropsis semialata, a unique grass species with both C(3) and C(4) subspecies. Experimental manipulation of rainfall in a common garden in South Africa tested the hypotheses that: (1) photosynthesis is greater in the C(4) than C(3) subspecies under high summer temperatures, but this pattern is reversed at low winter temperatures; and (2) the photosynthetic advantage of C(4) plants is enhanced during drought events. Measurements of leaf gas exchange over 2 years showed a significant photosynthetic advantage for the C(4) subspecies under irrigated conditions from spring through autumn. However, the C(4) leaves were killed by winter frost, while photosynthesis continued in the C(3) plants. Unexpectedly, the C(4) subspecies also lost its photosynthetic advantage during natural drought events, despite greater water-use efficiency under irrigated conditions. This study highlights previously unrecognized roles for climatic extremes in determining the ecological success of C(3) and C(4) grasses. PMID- 18410491 TI - Altered photosynthetic electron channelling into cyclic electron flow and nitrite assimilation in a mutant of ferredoxin:NADP(H) reductase. AB - The mechanism by which plants regulate channelling of photosynthetically derived electrons into different areas of chloroplast metabolism remains obscure. Possible fates of such electrons include use in carbon assimilation, nitrogen assimilation and redox signalling pathways, or return to the plastoquinone pool through cyclic electron flow. In higher plants, these electrons are made accessible to stromal enzymes, or for cyclic electron flow, as reduced ferredoxin (Fd), or NADPH. We investigated how knockout of an Arabidopsis (Arabidopsis thaliana) ferredoxin:NADPH reductase (FNR) isoprotein and the loss of strong thylakoid binding by the remaining FNR in this mutant affected the channelling of photosynthetic electrons into NADPH- and Fd-dependent metabolism. Chlorophyll fluorescence data show that these mutants have complex variation in cyclic electron flow, dependent on light conditions. Measurements of electron transport in isolated thylakoid and chloroplast systems demonstrated perturbed channelling to NADPH-dependent carbon and Fd-dependent nitrogen assimilating metabolism, with greater competition in the mutant. Moreover, mutants accumulate greater biomass than the wild type under low nitrate growth conditions, indicating that such altered chloroplast electron channelling has profound physiological effects. Taken together, our results demonstrate the integral role played by FNR isoform and location in the partitioning of photosynthetic reducing power. PMID- 18410492 TI - Photosynthesis in aquatic adventitious roots of the halophytic stem-succulent Tecticornia pergranulata (formerly Halosarcia pergranulata). AB - In flood-tolerant species, a common response to inundation is growth of adventitious roots into the water column. The capacity for these roots to become photosynthetically active has received scant attention. The experiments presented here show the aquatic adventitious roots of the flood-tolerant, halophytic stem succulent, Tecticornia pergranulata (subfamily Salicornioideae, Chenopodiaceae) are photosynthetic and quantify for the first time the photosynthetic capacity of aquatic roots for a terrestrial species. Fluorescence microscopy was used to determine the presence of chloroplasts within cells of aquatic roots. Net O(2) production by excised aquatic roots, when underwater, was measured with varying light and CO(2) regimes; the apparent maximum capacity (P(max)) for underwater net photosynthesis in aquatic roots was 0.45 micromol O(2) m(-2) s(-1). The photosynthetic potential of these roots was supported by the immunolocalization of PsbA, the major protein of photosystem II, and ribulose-1-5-bisphosphate carboxylase/oxygenase (Rubisco) in root protein extracts. Chlorophyllous aquatic roots of T. pergranulata are photosynthetically active, and such activity is a previously unrecognized source of O(2), and potentially carbohydrates, in flooded and submerged plants. PMID- 18410493 TI - Plasmodium falciparum Sec24 marks transitional ER that exports a model cargo via a diacidic motif. AB - Exit from the endoplasmic reticulum (ER) often occurs at distinct sites of vesicle formation known as transitional ER (tER) that are enriched for COPII vesicle coat proteins. We have characterized the organization of ER export in the malaria parasite, Plasmodium falciparum, by examining the localization of two components of the COPII machinery, PfSec12 and PfSec24a. PfSec12 was found throughout the ER, whereas the COPII cargo adaptor, PfSec24a, was concentrated at distinct foci that likely correspond to tER sites. These foci were closely apposed to cis-Golgi sites marked by PfGRASP-GFP, and upon treatment with brefeldin A they accumulated a model cargo protein via a process dependent on the presence of an intact diacidic export motif. Our data suggest that the cargo binding function of PfSec24a is conserved and that accumulation of cargo in discrete tER sites depends upon positive sorting signals. Furthermore, the number and position of tER sites with respect to the cis-Golgi suggests a co-ordinated biogenesis of these domains. PMID- 18410494 TI - The role of laccase in prostaglandin production by Cryptococcus neoformans. AB - Recently, it has been demonstrated that the opportunistic fungal pathogen Cryptococcus neoformans can synthesize authentic immunomodulatory prostaglandins. The mechanism by which this takes place is unclear as there is no cyclooxygenase homologue in the cryptococcal genome. In this study, we show that cryptococcal production of both PGE(2) and PGF(2 alpha) can be chemically inhibited by caffeic acid, resveratrol and nordihydroguaiaretic acid. These polyphenolic molecules are frequently used as inhibitors of lipoxygenase enzymes; however, blast searches of the cryptococcal genome were unable to identify any homologues of mammalian, plant or fungal lipoxygenases. Next we investigated cryptococcal laccase, an enzyme known to bind polyphenols, and found that either antibody depletion or genetic deletion of the primary cryptococcal laccase (lac1 Delta) resulted in a loss of cryptococcal prostaglandin production. To determine how laccase is involved, we tested recombinant laccase activity on the prostaglandin precursors, arachidonic acid (AA), PGG(2) and PGH(2). Using mass spectroscopy we determined that recombinant Lac1 does not modify AA or PGH(2), but does have a marked activity toward PGG(2) converting it to PGE(2) and 15-keto-PGE(2). These data demonstrate a critical role for laccase in cryptococcal prostaglandin production, and provides insight into a new and unique fungal prostaglandin pathway. PMID- 18410495 TI - The Ustilago maydis Cys2His2-type zinc finger transcription factor Mzr1 regulates fungal gene expression during the biotrophic growth stage. AB - The smut fungus Ustilago maydis establishes a biotrophic relationship with its host plant maize to progress through sexual development. Here, we report the identification and characterization of the Cys(2)His(2)-type zinc finger protein Mzr1 that functions as a transcriptional activator during host colonization. Expression of the U. maydis mig2 cluster genes is tightly linked to this phase. Upon conditional overexpression, Mzr1 confers induction of a subset of mig2 genes during vegetative growth and this requires the same promoter elements that confer inducible expression in planta. Furthermore, expression of the mig2-4 and mig2-5 genes during biotrophic growth is strongly reduced in cells deleted in mzr1. DNA array analysis led to the identification of additional Mzr1-induced genes. Some of these genes show a mig2-like plant-specific expression pattern and Mzr1 is responsible for their high-level expression during pathogenesis. Mzr1 function requires the b-dependently regulated Cys(2)His(2)-type cell cycle regulator Biz1, indicating that two stage-specific regulators mediate gene expression during host colonization. In spite of a role as transcriptional activator during biotrophic growth, mzr1 is not essential for pathogenesis; however, conditional overexpression interfered with proliferation during vegetative growth and mating ability, caused a cell separation defect, and triggered filamentous growth. We discuss the implications of these findings. PMID- 18410496 TI - Protein N-glycosylation determines functionality of the Saccharomyces cerevisiae cell wall integrity sensor Mid2p. AB - The fungal cell wall is a highly dynamic structure that is essential to maintain cell shape and stability. Hence in yeasts and fungi cell wall integrity is tightly controlled. The Saccharomyces cerevisiae plasma membrane protein Mid2p is a putative mechanosensor that responds to cell wall stresses and morphological changes during pheromone induction. The extracellular domain of Mid2p, which is crucial to sensing, is highly O- and N-glycosylated. We showed that O mannosylation is determining stability of Mid2p. If and how N-glycosylation is linked to Mid2p function was unknown. Here we demonstrate that Mid2p contains a single high mannose N-linked glycan at position Asn-35. The N-glycan is located close to the N-terminus and is exposed from the plasma membrane towards the cell wall through a highly O-mannosylated domain that is predicted to adopt a rod-like conformation. In contrast to O-mannosylation, lack of the N-linked glycan affects neither, stability of Mid2p nor distribution at the plasma membrane during vegetative and sexual growth. However, non-N-glycosylated Mid2p fails to perceive cell wall challenges. Our data further demonstrate that both the extent of the N linked glycan and its distance from the plasma membrane affect Mid2p function, suggesting the N-glycan to be directly involved in Mid2p sensing. PMID- 18410497 TI - DNA dynamics vary according to macrodomain topography in the E. coli chromosome. AB - The organization of the Escherichia coli chromosome has been defined genetically as consisting of four insulated macrodomains and two less constrained regions. Here we have examined the movement of chromosomal loci by tracking fluorescent markers in time-lapse microscopy during a complete cell cycle. Analysing the positioning, the segregation pattern and the motility of markers allowed us to show that the dynamic behaviour of loci belonging to various macrodomains and less constrained regions is radically different. In macrodomains constraints on mobility are apparent whereas in non-structured regions, markers exhibited a greater motility that may explain their ability to interact with flanking macrodomains. Following replication, duplicated markers belonging to macrodomains show a colocalization step and this landmark is not apparent in non-structured regions. Chromosome segregation occurs in three steps: first, the origin-proximal half of the chromosome consisting of the Ori macrodomain and the two non structured region segregates concomitantly in a short period of time. Second, the Right and Left macrodomains segregate progressively following the genetic map. Third, the Ter macrodomain is rapidly segregated before division, after a significant period of colocalization. Macrodomain territories defined as cellular spaces occupied by the different macrodomains can be identified. PMID- 18410498 TI - The Maurer's cleft protein MAHRP1 is essential for trafficking of PfEMP1 to the surface of Plasmodium falciparum-infected erythrocytes. AB - During the intra-erythrocytic development of Plasmodium falciparum, the parasite modifies the host cell surface by exporting proteins that interact with or insert into the erythrocyte membrane. These proteins include the principal mediator of cytoadherence, P. falciparum erythrocyte membrane protein 1 (PfEMP1). To implement these changes, the parasite establishes a protein-trafficking system beyond its confines. Membrane-bound structures called Maurer's clefts are intermediate trafficking compartments for proteins destined for the host cell membrane. We disrupted the gene for the membrane-associated histidine-rich protein 1 (MAHRP1). MAHRP1 is not essential for parasite viability or Maurer's cleft formation; however, in its absence, these organelles become disorganized in permeabilized cells. Maurer's cleft-resident proteins and transit cargo are exported normally in the absence of MAHRP1; however, the virulence determinant, PfEMP1, accumulates within the parasite, is depleted from the Maurer's clefts and is not presented at the red blood cell surface. Complementation of the mutant parasites with mahrp1 led to the reappearance of PfEMP1 on the infected red blood cell surface, and binding studies show that PfEMP1-mediated binding to CD36 is restored. These data suggest an important role of MAHRP1 in the translocation of PfEMP1 from the parasite to the host cell membrane. PMID- 18410499 TI - The essentiality and involvement of Streptococcus intermedius histone-like DNA binding protein in bacterial viability and normal growth. AB - Streptococcus intermedius histone-like DNA-binding protein (Si-HLP) is a homodimeric protein and, conserved with Escherichia coli HU, a well-documented nucleoid-associated protein (NAP). In E. coli, HU plays important roles as both structural and regulatory factors, but it is not essential for E. coli viability. Streptococcal HLP has been found to bind host cells and induce cytokine production, but its physiological role remains poorly defined. In the present study, using gene insertion knockout and tetracycline-regulated antisense RNA expression techniques, we determined whether Si-HLP is essential for bacterial viability and normal growth in S. intermedius. The Si-HLP-downregulated S. intermedius strain showed alterations in its morphology and surface properties. Downregulation of Si-HLP led to an expanded nucleoid to fill the intracellular space. Transcription levels of several genes, including virulence-associated factors, were found to be activated or repressed in the antisense Si-hlp RNA expressing strain by real-time PCR and reverse-transcription PCR. Collectively, these data suggest that Si-HLP serves as an essential NAP governing the nucleoid architecture and controlling the gene transcription profile in S. intermedius. PMID- 18410500 TI - Mitogen-activated protein kinase pathway mediates N-(4-hydroxyphenyl)retinamide induced neuronal differentiation in the ARPE-19 human retinal pigment epithelial cell line. AB - We have shown previously that N-(4-hydroxyphenyl)retinamide (4HPR, fenretinide), a retinoic acid derivative, induces neuronal differentiation in cultured human retinal pigment epithelial (RPE) cells [Chen et al., J. Neurochem., 84 (2003), 972]. We asked the question whether the mitogen-activated protein kinase (MAPK) pathway is involved in the regulation of the 4HPR-induced neuronal differentiation of RPE (ARPE-19) cells. When we treated ARPE-19 cells with 4HPR, c-Raf and MEK1/2 kinase were activated resulting in activation of the downstream effector ERK1/2 and of SAPK/JNK. By blocking the upstream kinase MEK1/2 with specific inhibitor U0126 we abrogated the 4HPR-induced phosphorylation of ERK1/2 and SAPK/JNK, indicating that the neuronal differentiation occurs through a positive cross-talk between the ERK and the SAPK/JNK pathways. Both U0126 and the suppression of ERK1/2 expression with small interfering RNA effectively blocked the 4HPR-induced neuronal differentiation of RPE cells and the expression of calretinin. The activated ERK1/2 then induced a sequential activation of p90RSK, and increase in phosphorylation of transcription factors c-fos and c-jun leading to transcriptional activation of AP-1. Taken together, our results clearly demonstrate that c-Raf/MEK1/2 signaling cascade involving ERK1/2 plays a central role in mediating the 4HPR-induced neuronal differentiation and calretinin expression in the human ARPE-19 retinal pigment epithelial cell line. PMID- 18410502 TI - Accumulation and clearance of alpha-synuclein aggregates demonstrated by time lapse imaging. AB - Aggregates of alpha-synuclein are the pathological hallmark of sporadic Parkinson's disease (PD), and mutations in the alpha-synuclein gene underlie familial forms of the disease. To characterize the formation of alpha-synuclein aggregates in living cells, we developed a new strategy to visualize alpha synuclein by fluorescence microscopy: alpha-synuclein was tagged with a six amino acid PDZ binding motif and co-expressed with the corresponding PDZ domain fused to enhanced green fluorescent protein (EGFP). In contrast to the traditional approach of alpha-synuclein-EGFP fusion proteins, this technique provided several fold higher sensitivity; this allowed us to compare alpha-synuclein variants and perform time-lapse imaging. A C-terminally truncated alpha-synuclein variant showed the highest prevalence of aggregates and toxicity, consistent with stabilization of the alpha-synuclein monomer by its C-terminus. Time-lapse imaging illustrated how cells form and accumulate aggregates of alpha-synuclein. A substantial number of cells also reduced their aggregate load, primarily through formation of an aggresome, which could itself be cleared from the cell. The molecular chaperone Hsp70 not only prevented the formation of aggregates, but also increased their reduction and clearance, underlining the therapeutic potential of similar strategies. In contrast to earlier assumptions build-up, reduction and clearance of alpha-synuclein aggregation thus appear a highly dynamic process. PMID- 18410501 TI - Cathepsin L participates in the production of neuropeptide Y in secretory vesicles, demonstrated by protease gene knockout and expression. AB - Neuropeptide Y (NPY) functions as a peptide neurotransmitter and as a neuroendocrine hormone. The active NPY peptide is generated in secretory vesicles by proteolytic processing of proNPY. Novel findings from this study show that cathepsin L participates as a key proteolytic enzyme for NPY production in secretory vesicles. Notably, NPY levels in cathepsin L knockout (KO) mice were substantially reduced in brain and adrenal medulla by 80% and 90%, respectively. Participation of cathepsin L in producing NPY predicts their colocalization in secretory vesicles, a primary site of NPY production. Indeed, cathepsin L was colocalized with NPY in brain cortical neurons and in chromaffin cells of adrenal medulla, demonstrated by immunofluorescence confocal microscopy. Immunoelectron microscopy confirmed the localization of cathepsin L with NPY in regulated secretory vesicles of chromaffin cells. Functional studies showed that coexpression of proNPY with cathepsin L in neuroendocrine PC12 cells resulted in increased production of NPY. Furthermore, in vitro processing indicated cathepsin L processing of proNPY at paired basic residues. These findings demonstrate a role for cathepsin L in the production of NPY from its proNPY precursor. These studies illustrate the novel biological role of cathepsin L in the production of NPY, a peptide neurotransmitter, and neuroendocrine hormone. PMID- 18410503 TI - Alpha-synuclein aggregation and cell death triggered by energy deprivation and dopamine overload are counteracted by D2/D3 receptor activation. AB - Progressive degeneration and intraneuronal Lewy bodies made of filamentous alpha synuclein (alpha-syn) in dopaminergic cells of the nigrostriatal system are characteristics of Parkinson's disease (PD). Glucose uptake is reduced in some of the brain regions affected by PD neurodegenerative changes. Defects in mitochondrial activity in the substantia nigra have been observed in the brain of patients affected by PD and substantia nigra lesions can induce the onset of a secondary parkinsonism. Thus, energy starvation and consequently metabolic impairment to dopaminergic neurons may be related to the onset of PD. On this line, we evaluated the effect of nutrient starvation, reproduced 'in vitro' by glucose deprivation (GD), in primary mesecephalic neuronal cultures and dopaminergic-differentiated SH-SY5Y cells, to evaluate if decreased glucose support to dopaminergic cells can lead to mitochondrial damage, neurodegeneration and alpha-syn misfolding. Furthermore, we investigated the effect of dopamine (DA) treatment in the presence of a DA-uptake inhibitor or of the D(2)/D(3) receptor (D(2)R/D(3)R) agonist quinpirole on GD-treated cells, to evaluate the efficacy of these therapeutic compounds. We found that GD induced the formation of fibrillary aggregated alpha-syn inclusions containing the DA transporter in dopaminergic cells. These alterations were accompanied by dopaminergic cell death and were exacerbated by DA overload. Conversely, the block of DA uptake and D(2)R/D(3)R agonist treatment exerted neuroprotective effects. These data indicate that glucose starvation is likely involved in the induction of PD related pathological changes in dopaminergic neurons. These changes may be counteracted by the block of DA uptake and by dopaminergic agonist treatment. PMID- 18410505 TI - Cerebral ischemia/stroke and small ubiquitin-like modifier (SUMO) conjugation--a new target for therapeutic intervention? AB - Transient cerebral ischemia/stroke activates various post-translational protein modifications such as phosphorylation and ubiquitin conjugation that are believed to play a major role in the pathological process triggered by an interruption of blood supply and culminating in cell death. A new system of post-translational protein modification has been identified, termed as small ubiquitin-like modifier (SUMO) conjugation. Like ubiquitin, SUMO is conjugated to the lysine residue of target proteins in a complex process. This review summarizes observations from recent experiments focusing on the effect of cerebral ischemia on SUMO conjugation. Transient global and focal cerebral ischemia both induced a rapid, dramatic and long-lasting rise in levels of SUMO2/3 conjugation. After transient focal cerebral ischemia, SUMO conjugation was particularly prominent in neurons located at the border of the ischemic territory where SUMO-conjugated proteins translocated to the nucleus. Many SUMO conjugation target proteins are transcription factors and sumoylation has been shown to have a major impact on the activity, stability, and cellular localization of target proteins. The rise in levels of SUMO-conjugated proteins is therefore likely to have a major effect on the fate of post-ischemic neurons. The sumoylation process could provide an exciting new target for therapeutic intervention. PMID- 18410504 TI - Glial connexins and gap junctions in CNS inflammation and disease. AB - Gap junctions facilitate direct cytoplasmic communication between neighboring cells, facilitating the transfer of small molecular weight molecules involved in cell signaling and metabolism. Gap junction channels are formed by the joining of two hemichannels from adjacent cells, each composed of six oligomeric protein subunits called connexins. Of paramount importance to CNS homeostasis are astrocyte networks formed by gap junctions, which play a critical role in maintaining the homeostatic regulation of extracellular pH, K+, and glutamate levels. Inflammation is a hallmark of several diseases afflicting the CNS. Within the past several years, the number of publications reporting effects of cytokines and pathogenic stimuli on glial gap junction communication has increased dramatically. The purpose of this review is to discuss recent observations characterizing the consequences of inflammatory stimuli on homocellular gap junction coupling in astrocytes and microglia as well as changes in connexin expression during various CNS inflammatory conditions. PMID- 18410506 TI - Poly (ADP-ribose) polymerases (PARPs) 1-3 regulate astrocyte activation. AB - Besides their traditional role in maintaining CNS homeostasis, astrocytes also participate in innate immune responses. Indeed, we have previously demonstrated that astrocytes are capable of recognizing bacterial pathogens such as Staphylococcus aureus, a common etiologic agent of CNS infections, and respond with the robust production of numerous proinflammatory mediators. Suppression of Poly (ADP-ribose) polymerase-1 (PARP-1), a DNA repair enzyme, has been shown to attenuate inflammatory responses in several cell types including mixed glial cultures. However, a role for PARP-1 in regulating innate immune responses in purified astrocytes and the potential for multiple PARP family members to cooperatively regulate astrocyte activation has not yet been examined. The synthetic PARP-1 inhibitor PJ-34 attenuated the production of several proinflammatory mediators by astrocytes in response to S. aureus stimulation including nitric oxide, interleukin-1 beta, tumor necrosis factor-alpha, and CCL2. The release of all four mediators was partially reduced in PARP-1 knockout (KO) astrocytes compared to wild-type cells. The residual inflammatory mediator expression detected in PARP-1 KO astrocytes was further blocked with PJ-34, suggesting either non-specific effects of the drug or actions on alternative PARP isoforms. Reduction in PARP-2 or PARP-3 expression by siRNA knock down revealed that these isoforms also contributed to inflammatory mediator regulation in response to S. aureus. Interestingly, the combined targeting of either PARP 1/PARP-2 or PARP-2/PARP-3 attenuated astrocyte inflammatory responses more effectively compared to knock down of either PARP alone, suggesting cooperativity between PARP isoforms. Collectively, these findings suggest that PARPs influence the extent of S. aureus-induced astrocyte activation. PMID- 18410507 TI - Human brain aminopeptidase A: biochemical properties and distribution in brain nuclei. AB - Aminopeptidase A (APA) generated brain angiotensin III, one of the main effector peptides of the brain renin angiotensin system, exerting a tonic stimulatory effect on the control of blood pressure in hypertensive rats. The distribution of APA in human brain has not been yet studied. We first biochemically characterized human brain APA (apparent molecular mass of 165 and 130 kDa) and we showed that the human enzyme exhibited similar enzymatic characteristics to recombinant mouse APA. Both enzymes had similar sensitivity to Ca(2+). Kinetic studies showed that the K(m) (190 mumol/L) of the human enzyme for the synthetic substrate-l-glutamyl beta-naphthylamide was close from that of the mouse enzyme (256 mumol/L). Moreover, various classes of inhibitors including the specific and selective APA inhibitor, (S)-3-amino-4-mercapto-butyl sulfonic acid, had similar inhibitory potencies toward both enzymes. Using (S)-3-amino-4-mercapto-butyl sulfonic acid, we then specifically measured the activity of APA in 40 microdissected areas of the adult human brain. Significant heterogeneity was found in the activity of APA in the various analyzed regions. The highest activity was measured in the choroids plexus and the pineal gland. High activity was also detected in the dorsomedial medulla oblongata, in the septum, the prefrontal cortex, the olfactory bulb, the nucleus accumbens, and the hypothalamus, especially in the paraventricular and supraoptic nuclei. Immunostaining of human brain sections at the level of the medulla oblongata strengthened these data, showing for the first time a high density of immunoreactive neuronal cell bodies and fibers in the motor hypoglossal nucleus, the dorsal motor nucleus of the vagus, the nucleus of the solitary tract, the Roller nucleus, the ambiguus nucleus, the inferior olivary complex, and in the external cuneate nucleus. APA immunoreactivity was also visualized in vessels and capillaries in the dorsal motor nucleus of the vagus and the inferior olivary complex. The presence of APA in several human brain nuclei sensitive to angiotensins and involved in blood pressure regulation suggests that APA in humans is an integral component of the brain renin angiotensin system and strengthens the idea that APA inhibitors could be clinically tested as an additional therapy for the treatment of certain forms of hypertension. PMID- 18410508 TI - Methamphetamine-induced neurotoxicity and microglial activation are not mediated by fractalkine receptor signaling. AB - Methamphetamine (METH) damages dopamine (DA) nerve endings by a process that has been linked to microglial activation but the signaling pathways that mediate this response have not yet been delineated. Cardona et al. [Nat. Neurosci. 9 (2006), 917] recently identified the microglial-specific fractalkine receptor (CX3CR1) as an important mediator of MPTP-induced neurodegeneration of DA neurons. Because the CNS damage caused by METH and MPTP is highly selective for the DA neuronal system in mouse models of neurotoxicity, we hypothesized that the CX3CR1 plays a role in METH-induced neurotoxicity and microglial activation. Mice in which the CX3CR1 gene has been deleted and replaced with a cDNA encoding enhanced green fluorescent protein (eGFP) were treated with METH and examined for striatal neurotoxicity. METH depleted DA, caused microglial activation, and increased body temperature in CX3CR1 knockout mice to the same extent and over the same time course seen in wild-type controls. The effects of METH in CX3CR1 knockout mice were not gender-dependent and did not extend beyond the striatum. Striatal microglia expressing eGFP constitutively show morphological changes after METH that are characteristic of activation. This response was restricted to the striatum and contrasted sharply with unresponsive eGFP-microglia in surrounding brain areas that are not damaged by METH. We conclude from these studies that CX3CR1 signaling does not modulate METH neurotoxicity or microglial activation. Furthermore, it appears that striatal-resident microglia respond to METH with an activation cascade and then return to a surveying state without undergoing apoptosis or migration. PMID- 18410509 TI - Enhanced expression of Harvey ras induced by serum deprivation in cultured astrocytes. AB - Trophic deprivation contributes to astrocyte damage that occurs in acute and chronic neurodegenerative disorders. Unraveling the underlying mechanisms may pave way to novel cytoprotective strategies. Cultured mouse astrocytes responded to trophic deprivation with a large and transient increase in the expression of p21(ras), which was secondary to an enhanced formation of reactive oxygen species (ROS) detected by cytofluorimetric analysis after preloading with 2',7' dichlorofluorescein diacetate. The increase in p21(ras) levels was largely attenuated by the reducing agent, N-acetylcysteine, which was proven to reduce ROS formation in astrocytes subjected to serum deprivation. We extended the analysis to the Ha-Ras isoform, which has been implicated in mechanisms of cytotoxicity. We found that serum deprivation enhanced the expression and activity of Ha-Ras without changing Ha-Ras mRNA levels. The increase in Ha-Ras levels was sensitive to the protein synthesis inhibitor, cycloheximide, suggesting that serum deprivation increases translation of preformed Ha-Ras mRNA. The late decline in Ha-Ras levels observed after 60 min was prevented by the proteasome inhibitor, MG132, as well as by the selective mitogen-activated protein kinase (MAPK) inhibitor, PD98059. Serum deprivation led to the activation of the MAPK pathway in cultured astrocytes, as shown by an increase in phosphorylated extracellular signal-regulated kinase 1/2 levels after 5 and 30 min. Finally, using the siRNA technology, we found that an acute knock-down of Ha Ras was protective against astrocyte damage induced by serum deprivation. We conclude that cultured astrocytes respond to trophic deprivation with an increased expression in Ha-Ras, which is limited by the concomitant activation of the MAPK pathway, but is nevertheless involved in the pathophysiology of cell damage. PMID- 18410511 TI - Altered expression of key dopaminergic regulatory proteins in the postnatal brain following perinatal n-3 fatty acid dietary deficiency. AB - The consequences of maternal linolenic acid (LNA, 18:3n-3) dietary deficiency on key dopamine (DA)-associated regulatory proteins in mesolimbic and mesocortical structures of the postnatal rat brain have been investigated. A marked (4.5-fold) decrease of the DA-synthesizing enzyme tyrosine hydroxylase accompanied by a down regulation (approx 7.5-fold) of the vesicular monoamine transporter (VMAT-2) and a depletion of VMAT-associated vesicles in the hippocampus were observed in deficient offspring compared with adequately fed controls. The DA transporter (DAT) was not affected by the LNA deficiency indicative of a DAT/VMAT-2 ratio increase that may enhance the risk of damage of the dopaminergic (DAergic) terminal. A robust increase in DA receptor (DAR1 and DAR2) levels was noticed in the cortex and striatum structures possibly to compensate for the low levels of DA in synaptic clefts. Microglia activation characterized by enhanced levels of ED1 antibody and nuclear internalization of p65 NFkappaB was noticed following LNA deficiency. Diminished levels of 22:6n-3 docosahexaenoic acid (Schiefermeier and Yavin 2002), the most ubiquitous metabolite generated by LNA is proposed to reduce the anti-oxidant arsenal in the developing brain and cause microglia activation and enhanced oxidative stress to increase the risk of certain DA associated neurological disorders. PMID- 18410510 TI - Intra-neural administration of fractalkine attenuates neuropathic pain-related behaviour. AB - There is increasing evidence that a number of cytokines and their receptors are involved in the processes that lead to the development and maintenance of neuropathic pain states. Here we demonstrate that levels of CX3CR1 (the receptor for the chemokine fractalkine) mRNA in lumbar dorsal root ganglia (DRG) increase 5.8-fold 7 days after sciatic nerve axotomy, and 1.7- and 2.9-fold, 3 and 7 days respectively, after the spared nerve injury (SNI) model of neuropathic pain. In contrast, no significant change in the levels of fractalkine mRNA is apparent in the DRG after axotomy or SNI. The increase in CX3CR1 mRNA is paralleled by a 3.9- and 2.1-fold increase in the number of CX3CR1-positive macrophages in the DRG 7 days after axotomy and SNI, respectively. Expression of CX3CR1 in macrophages is also markedly increased in the sciatic nerve proximal to site of injury, by 25.7 fold after axotomy and 16.2-fold after SNI, 7 days after injury. Intra-neural injection into the sciatic nerve of 400 ng or 100 ng of fractalkine in adult 129OlaHsd mice significantly delayed the development of allodynia for 3 days following SNI. Further, CX3CR1 knockout (KO) mice display an increase in allodynia for three weeks after SNI compared to strain-matched Balb/c controls. Taken together, these results suggest an anti-allodynic role for fractalkine and its receptor in the mouse. PMID- 18410512 TI - Striatal histone modifications in models of levodopa-induced dyskinesia. AB - Despite recent advances in the treatment of Parkinson disease (PD), levodopa remains the most effective and widely used therapy. A major limitation to the use of levodopa is the development of abnormal involuntary movements, termed levodopa induced dyskinesia (LDID), following chronic levodopa treatment. Since recent studies have suggested that modifications of chromatin structure may be responsible for many long-lasting changes in brain function, we have examined post-translational modifications of striatal histones in two models of LDID: an acute murine model and a chronic macaque monkey model, both exposed to 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP). In the primate model, which closely resembles human LDID, we observed that chronic levodopa and the appearance of LDID was associated with marked deacetylation of histone H4, hyperacetylation and dephosphorylation of histone H3, and enhancement of the phosphorylation of extracellular signal-regulated kinase (ERK). In the murine model of acutely rather than chronically induced LDID, dopamine depletion and levodopa treatment also induced deacetylation of histone H4 and phosphorylation of ERK, but histone H3 exhibited decreased trimethylation and reduced rather than enhanced acetylation. These data demonstrate striking changes in striatal histones associated with the induction of LDID in both animal models. The pattern of changes observed, as well as the behavioral features, differed in the two models. However, both models exhibit marked deacetylation of histone H4, suggesting that inhibitors of H4 deacetylation may be useful in preventing or reversing LDID. PMID- 18410513 TI - High cholesterol-induced neuroinflammation and amyloid precursor protein processing correlate with loss of working memory in mice. AB - Recent findings suggest that hypercholesterolemia may contribute to the onset of Alzheimer's disease-like dementia but the underlying mechanisms remain unknown. In this study, we evaluated the cognitive performance in rodent models of hypercholesterolemia in relation to neuroinflammatory changes and amyloid precursor protein (APP) processing, the two key parameters of Alzheimer's disease pathogenesis. Groups of normal C57BL/6 and low density lipoprotein receptor (LDLR)-deficient mice were fed a high fat/cholesterol diet for an 8-week period and tested for memory in a radial arm maze. It was found that the C57BL/6 mice receiving a high fat diet were deficient in handling an increasing working memory load compared with counterparts receiving a control diet while the hypercholesterolemic LDLR-/- mice showed impaired working memory regardless of diet. Immunohistochemical analysis revealed the presence of activated microglia and astrocytes in the hippocampi from high fat-fed C57BL/6 mice and LDLR-/- mice. Consistent with a neuroinflammatory response, the hyperlipidemic mice showed increased expression of cytokines/mediators including tumor necrosis factor alpha, interleukin-1beta and -6, nitric oxide synthase 2, and cycloxygenase 2. There was also an induced expression of the key APP processing enzyme i.e. beta site APP cleaving enzyme 1 in both high fat/cholesterol-fed C57BL/6 and LDLR-/- mice accompanied by an increased generation of C-terminal fragments of APP. Although ELISA for beta-amyloid failed to record significant changes in the non transgenic mice, a threefold increase in beta-amyloid 40 accumulation was apparent in a strain of transgenic mice expressing wild-type human APP on high fat/cholesterol diet. The findings link hypercholesterolemia with cognitive dysfunction potentially mediated by increased neuroinflammation and APP processing in a non-transgenic mouse model. PMID- 18410514 TI - Spastin oligomerizes into a hexamer and the mutant spastin (E442Q) redistribute the wild-type spastin into filamentous microtubule. AB - Spastin, a member of the ATPases associated with various cellular activities (AAA) family of proteins, is the most frequently mutated in hereditary spastic paraplegia. The defining feature of the AAA proteins is a structurally conserved AAA domain which assembles into an oligomer. By chemical cross-linking and gel filtration chromatography, we show that spastin oligomerizes into a hexamer. Furthermore, to gain a comprehensive overview of the oligomeric structure of spastin, we generated a structural model of the AAA domain of spastin using template structure of VPS4B and p97/VCP. The generated model of spastin provided us with a framework to classify the identified missense mutations in the AAA domain from hereditary spastic paraplegia patients into different structural/functional groups. Finally, through co-localization studies in mammalian cells, we show that E442Q mutant spastin acts in a dominant negative fashion and causes redistribution of both wild-type spastin monomer and spastin interacting protein, RTN1 into filamentous microtubule bundles. PMID- 18410516 TI - Mechanisms of non-steroid anti-inflammatory drugs action on ASICs expressed in hippocampal interneurons. AB - The inhibitory action of non-steroid anti-inflammatory drugs was investigated on acid-sensing ionic channels (ASIC) in isolated hippocampal interneurons and on recombinant ASICs expressed in Chinese hamster ovary (CHO) cells. Diclofenac and ibuprofen inhibited proton-induced currents in hippocampal interneurons (IC(50) were 622 +/- 34 muM and 3.42 +/- 0.50 mM, respectively). This non-competitive effect was fast and fully reversible for both drugs. Aspirin and salicylic acid at 500 muM were ineffective. Diclofenac and ibuprofen decreased the amplitude of proton-evoked currents and slowed the rates of current decay with a good correlation between these effects. Simultaneous application of acid solution and diclofenac was required for its inhibitory effect. Unlike amiloride, the action of diclofenac was voltage-independent and no competition between two drugs was found. Analysis of the action of diclofenac and ibuprofen on activation and desensitization of ASICs showed that diclofenac but not ibuprofen shifted the steady-state desensitization curve to more alkaline pH values. The reason for this shift was slowing down the recovery from desensitization of ASICs. Thus, diclofenac may serve as a neuroprotective agent during pathological conditions associated with acidification. PMID- 18410518 TI - Region-selective alterations of glucose oxidation and amino acid synthesis in the thiamine-deficient rat brain: a re-evaluation using 1H/13C nuclear magnetic resonance spectroscopy. AB - Thiamine deficiency provides an effective model of selective neuronal cell death. (1)H and (13)C-NMR was used to investigate the effects of thiamine deficiency on the synthesis of amino acids derived from [1-(13)C]glucose in vulnerable (medial thalamus; MT) compared to non-vulnerable (frontal cortex; FC) brain regions. Following 11 days of thiamine deficiency, a time-point associated with the absence of significant neuronal cell death, regional concentrations of glutamate, glutamine and GABA remained unaffected in FC and MT; however, decreased levels of aspartate in MT at this time-point were a predictor of regional vulnerability. De novo synthesis of glutamate and GABA were unaffected at 11 days of thiamine deficiency, while synthesis of [2-(13)C]aspartate was significantly impaired. Glucose loading, which has been shown to exacerbate symptoms in patients with thiamine deficiency, resulted in further decreases of TCA cycle flux and reduced de novo synthesis of glutamate, aspartate and GABA in thiamine-deficient (TD) rats. Isotopomer analysis revealed that impaired TCA cycle flux and decreased aspartate synthesis due to thiamine deficiency occurred principally in neurons. Glucose loading deteriorated TD-related decreases in TCA cycle flux, and concomitantly reduced synthesis of aspartate and glutamate in MT. PMID- 18410515 TI - Expression of microRNAs and their precursors in synaptic fractions of adult mouse forebrain. AB - We have characterized the expression of microRNAs and selected microRNA precursors within several synaptic fractions of adult mouse forebrain, including synaptoneurosomes, synaptosomes and isolated post-synaptic densities (PSDs), using methods of microRNA microarray, real time qRT-PCR, Northern blotting and immunopurification using anti-PSD95 antibody. The majority of brain microRNAs (especially microRNAs known to be expressed in pyramidal neurons) are detectably expressed in synaptic fractions, and a subset of microRNAs is significantly enriched in synaptic fractions relative to total forebrain homogenate. MicroRNA precursors are also detectable in synaptic fractions at levels that are comparable to whole tissue. Whereas mature microRNAs are predominantly associated with soluble components of the synaptic fractions, microRNA precursors are predominantly associated with PSDs. For seven microRNAs examined, there was a significant correlation between the relative synaptic enrichment of the precursor and the relative synaptic enrichment of the corresponding mature microRNA. These findings support the proposal that microRNAs are formed, at least in part, via processing of microRNA precursors locally within dendritic spines. Dicer is expressed in PSDs but is enzymatically inactive until conditions that activate calpain cause its liberation; thus, we propose that synaptic stimulation may lead to local processing of microRNA precursors in proximity to the synapse. PMID- 18410517 TI - FasL shedding is reduced by hypothermia in experimental stroke. AB - Protection by mild hypothermia has previously been associated with better mitochondrial preservation and suppression of the intrinsic apoptotic pathway. It is also known that the brain may undergo apoptotic death via extrinsic, or receptor-mediated pathways, such as that triggered by Fas/FasL. Male Sprague Dawley rats subjected to 2 h middle cerebral artery occlusion with 2 h intraischemic mild hypothermia (33 degrees C) were assayed for Fas, FasL and caspase-8 expression. Ischemia increased Fas, but decreased FasL by approximately 50-60% at 6 and 24 h post-insult. Mild hypothermia significantly reduced expression of Fas and processed caspase-8 both by approximately 50%, but prevented ischemia-induced FasL decreases. Fractionation revealed that soluble/shed FasL (sFasL) was decreased by hypothermia, while membrane-bound FasL (mFasL) increased. To more directly assess the significance of the Fas/FasL pathway in ischemic stroke, primary neuron cultures were exposed to oxygen glucose deprivation. Since FasL is cleaved by matrix metalloproteinases (MMPs), and mild hypothermia decreases MMP expression, treatment with a pan-MMP inhibitor also decreased sFasL. Thus, mild hypothermia is associated with reduced Fas expression and caspase-8 activation. Hypothermia prevented total FasL decreases, and most of it remained membrane-bound. These findings reveal new observations regarding the effect of mild hypothermia on the Fas/FasL and MMP systems. PMID- 18410519 TI - Pre-synaptic and post-synaptic localization of EphA4 and EphB2 in adult mouse forebrain. AB - The ephrin receptors EphA4 and EphB2 have been implicated in synaptogenesis and long-term potentiation in the cerebral cortex and hippocampus, where they are generally viewed as post-synaptic receptors. To determine the precise distribution of EphA4 and EphB2 in mature brain synapses, we used subcellular fractionation and electron microscopy to examine the adult mouse forebrain/midbrain. EphA4 and EphB2 were both enriched in microsomes and synaptosomes. In synaptosomes, they were present in the membrane and the synaptic vesicle fractions. While EphA4 was tightly associated with PSD-95-enriched post synaptic density fractions, EphB2 was easily extracted with detergents. In contrast, both receptors were found in the pre-synaptic active zone fraction. By electron microscopy, EphA4 was mainly detected in axon terminals, whereas EphB2 was more frequently detected in large dendritic shafts, in the hippocampus and cerebral cortex. However, in the ventrobasal thalamus, EphB2 was detected most frequently in axon terminals and thin dendritic shafts. The localization of EphA4 and EphB2 in multiple compartments of neurons and synaptic junctions suggests that they interact with several distinct scaffolding proteins and play diverse roles at synapses. PMID- 18410520 TI - Electron tomography and immunonanogold electron microscopy for investigating intracellular trafficking and secretion in human eosinophils. AB - Electron tomography (ET) has increasingly been used to understand the complexity of membrane systems and protein-trafficking events. By ET and immunonanogold electron microscopy, we recently defined a route for vesicular transport and release of granule-stored products from within activated human eosinophils, cells specialized in the secretion of numerous cytokines and other proteins during inflammatory responses. Here, we highlight these techniques as important tools to unveil a distinct eosinophil vesicular system and secretory pathway. PMID- 18410521 TI - Frabin and other related Cdc42-specific guanine nucleotide exchange factors couple the actin cytoskeleton with the plasma membrane. AB - Frabin, together with, at least, FGD1, FGD2, FGD3 and FGD1-related Cdc42-GEF (FRG), is a member of a family of Cdc42-specific gua-nine nucleotide exchange factors (GEFs). These proteins have multiple phosphoinositide-binding domains, including two pleckstrin homology (PH) domains and an FYVE or FERM domain. It is likely that they couple the actin cytoskeleton with the plasma membrane. Frabin associates with a specific actin structure(s) and induces the direct activation of Cdc42 in the vicinity of this structure(s), resulting in actin reorganization. Furthermore, frabin associates with a specific membrane structure(s) and induces the indirect activation of Rac in the vicinity of this structure(s), resulting in the reorganization of the actin cytoskeleton. This reorganization of the actin cytoskeleton induces cell shape changes such as the formation of filopodia and lamellipodia. PMID- 18410522 TI - Flavonoid-mediated presenilin-1 phosphorylation reduces Alzheimer's disease beta amyloid production. AB - Glycogen synthase kinase 3 (GSK-3) dysregulation is implicated in the two Alzheimer's disease (AD) pathological hallmarks: beta-amyloid plaques and neurofibrillary tangles. GSK-3 inhibitors may abrogate AD pathology by inhibiting amyloidogenic gamma-secretase cleavage of amyloid precursor protein (APP). Here, we report that the citrus bioflavonoid luteolin reduces amyloid-beta (Abeta) peptide generation in both human 'Swedish' mutant APP transgene-bearing neuron like cells and primary neurons. We also find that luteolin induces changes consistent with GSK-3 inhibition that (i) decrease amyloidogenic gamma-secretase APP processing, and (ii) promote presenilin-1 (PS1) carboxyl-terminal fragment (CTF) phosphorylation. Importantly, we find GSK-3alpha activity is essential for both PS1 CTF phosphorylation and PS1-APP interaction. As validation of these findings in vivo, we find that luteolin, when applied to the Tg2576 mouse model of AD, decreases soluble Abeta levels, reduces GSK-3 activity, and disrupts PS1 APP association. In addition, we find that Tg2576 mice treated with diosmin, a glycoside of a flavonoid structurally similar to luteolin, display significantly reduced Abeta pathology. We suggest that GSK-3 inhibition is a viable therapeutic approach for AD by impacting PS1 phosphorylation-dependent regulation of amyloidogenesis. PMID- 18410523 TI - Imaging of primary human hepatocytes performed with micron-sized iron oxide particles and clinical magnetic resonance tomography. AB - Transplantation of primary human hepatocytes is a promising approach in certain liver diseases. For the visualization of the hepatocytes during and following cell application and the ability of a timely response to potential complications, a non-invasive modality for imaging the transplanted cells has to be established. The aim of this study was to label primary human hepatocytes with micron-sized iron oxide particles (MPIOs), enabling the detection of cells by clinical magnetic resonance imaging (MRI). Primary human hepatocytes isolated from 13 different donors were used for the labelling experiments. Following the dose finding studies, hepatocytes were incubated with 30 particles/cell for 4 hrs in an adhesion culture. Particle incorporation was investigated via light, fluorescence and electron microscopy, and labelled cells were fixed and analysed in an agarose suspension by a 3.0 Tesla MR scanner. The hepatocytes were enzymatically resuspended and analysed during a 5-day reculture period for viability, total protein, enzyme leakage (aspartate aminotransferase [AST], lactate dehydrogenase [LDH]) and metabolic activity (urea, albumin). A mean uptake of 18 particles/cell could be observed, and the primary human hepatocytes were clearly detectable by MR instrumentation. The particle load was not affected by resuspension and showed no alternations during the culture period. Compared to control groups, labelling and resuspension had no adverse effects on the viability, enzyme leakage and metabolic activity of the human hepatocytes. The feasibility of preparing MPIO-labelled primary human hepatocytes detectable by clinical MR equipment was shown in vitro. MPIO-labelled cells could serve for basic research and quality control in the clinical setting of human hepatocyte transplantation. PMID- 18410524 TI - Mitochondrial nutrients improve immune dysfunction in the type 2 diabetic Goto Kakizaki rats. AB - The development of type 2 diabetes is accompanied by decreased immune function and the mechanisms are unclear. We hypothesize that oxidative damage and mitochondrial dysfunction may play an important role in the immune dysfunction in diabetes. In the present study, we investigated this hypothesis in diabetic Goto Kakizaki rats by treatment with a combination of four mitochondrial-targeting nutrients, namely, R-alpha-lipoic acid, acetyl-L-carnitine, nicotinamide and biotin. We first studied the effects of the combination of these four nutrients on immune function by examining cell proliferation in immune organs (spleen and thymus) and immunomodulating factors in the plasma. We then examined, in the plasma and thymus, oxidative damage biomarkers, including lipid peroxidation, protein oxidation, reactive oxygen species, calcium and antioxidant defence systems, mitochondrial potential and apoptosis-inducing factors (caspase 3, p53 and p21). We found that immune dysfunction in these animals is associated with increased oxidative damage and mitochondrial dysfunction and that the nutrient treatment effectively elevated immune function, decreased oxidative damage, enhanced mitochondrial function and inhibited the elevation of apoptosis factors. These effects are comparable to, or greater than, those of the anti-diabetic drug pioglitazone. These data suggest that a rational combination of mitochondrial targeting nutrients may be effective in improving immune function in type 2 diabetes through enhancement of mitochondrial function, decreased oxidative damage, and delayed cell death in the immune organs and blood. PMID- 18410525 TI - Development of a novel antimicrobial peptide, AG-30, with angiogenic properties. AB - The utility of various synthetic peptides has been investigated in clinical trials of the treatment of cancers, infectious diseases and endocrine diseases. In the process of functional gene screening with in silico analysis for molecules with angiogenic properties, we generated a small peptide, angiogenic peptide (AG) 30, that possesses both antimicrobial and pro-inflammatory activities. AG-30 has an alpha-helix structure with a number of hydrophobic or net positively charged amino acids and a propensity to fold into amphipathic structures. Indeed, AG-30 exhibited antimicrobial activity against various bacteria, induced vascular endothelial cell growth and tube formation in a dose-dependent manner and increased neovascularization in a Matrigel plug assay. As a result, AG-30 up regulated expression of angiogenesis-related cytokines and growth factors for up to 72 hrs in human aortic endothelial cells. To further evaluate the angiogenic effect of AG-30 in vivo, we developed a slow-release AG-30 system utilizing biodegradable gelatin microspheres. In the ischaemic mouse hind limb, slow release AG-30 treatment results in an increase in angiogenic score, an increase in blood flow (as demonstrated by laser Doppler imaging) and an increase in capillary density (as demonstrated by immunostaining with anti-CD31 antibody). These data suggest that the novel peptide, AG-30, may have therapeutic potential for ischaemic diseases. PMID- 18410526 TI - Endothelial cells from cord blood CD133+CD34+ progenitors share phenotypic, functional and gene expression profile similarities with lymphatics. AB - The existence of endothelial progenitor cells (EPC) with high cell-cycle rate in human umbilical cord blood has been recently shown and represents a challenging strategy for therapeutic neovascularization. To enhance knowledge for future cellular therapy, we compared the phenotypic, functional and gene expression differences between EPC-derived cells generated from cord blood CD34(+) cells, and lymphatic and macrovascular endothelial cells (EC) isolated from human foreskins and umbilical veins, respectively. Under appropriate culture conditions, EPC developed into fully matured EC with expression of similar endothelial markers as lymphatic and macrovascular EC, including CD31, CD36, von Willebrand factor FVIII, CD54 (ICAM-1), CD105 (endoglin), CD144 (VE-cadherin), Tie-1, Tie-2, VEGFR-1/Flt-1 and VEGFR-2/Flk-1. Few EPC-derived cells became positive for LYVE-1, indicating their origin from haematopoietic stem cells. However they lacked expression of other lymphatic cell-specific markers such as podoplanin and Prox-1. Functional tests demonstrated that the cobblestone EPC derived cells up-regulated CD54 and CD62E expression in response to TNF-alpha, incorporated DiI-acetylated low-density liproprotein and formed cord- and tubular like structures with capillary lumen in three-dimensional collagen culture--all characteristic features of the vascular endothelium. Structures compatible with Weibel-Palade bodies were also found by electron microscopy. Gene microarray profiling revealed that only a small percentage of genes investigated showed differential expression in EPC-derived cells and lymphatic EC. Among them were adhesion molecules, extracellular matrix proteins and cytokines. Our data point to the close lineage relationship of both types of vascular cells and support the theory of a venous origin of the lymphatic system. PMID- 18410527 TI - Curcumin-induced fibroblast apoptosis and in vitro wound contraction are regulated by antioxidants and heme oxygenase: implications for scar formation. AB - Fibroblast apoptosis plays a crucial role in normal and pathological scar formation and therefore we studied whether the putative apoptosis-inducing factor curcumin affects fibroblast apoptosis and may function as a novel therapeutic. We show that 25-microM curcumin causes fibroblast apoptosis and that this could be inhibited by co-administration of antioxidants N-acetyl-l-cysteine (NAC), biliverdin or bilirubin, suggesting that reactive oxygen species (ROS) are involved. This is supported by our observation that 25-microM curcumin caused the generation of ROS, which could be completely blocked by addition of NAC or bilirubin. Since biliverdin and bilirubin are downstream products of heme degradation by heme oxygenase (HO), it has been suggested that HO-activity protects against curcumin-induced apoptosis. Interestingly, exposure to curcumin maximally induced HO-1 protein and HO-activity at 10-15 microM, whereas, at a concentration of >20-microM curcumin HO-1-expression and HO-activity was negligible. NAC-mediated inhibition of 25-microM curcumin-induced apoptosis was demonstrated to act in part via restored HO-1-induction, since the rescuing effect of NAC could be reduced by inhibiting HO-activity. Moreover pre-induction of HO-1 using 5-microM curcumin protected fibroblasts against 25-microM curcumin induced apoptosis. On a functional level, fibroblast-mediated collagen gel contraction, an in vitro wound contraction model, was completely prevented by 25 microM curcumin, while this could be reversed by co-incubation with NAC, an effect that was also partially HO-mediated. In conclusion, curcumin treatment in high doses (>25 microM) may provide a novel way to modulate pathological scar formation through the induction of fibroblast apoptosis, while antioxidants, HO activity and its effector molecules act as a possible fine-tuning regulator. PMID- 18410528 TI - Endothelial cell differentiation of human breast tumour stem/progenitor cells. AB - Breast tumour stem cells have been reported to differentiate in the epithelial lineage but a cross-lineage potential has not been investigated. We aimed to evaluate whether breast tumour stem cells were able to differentiate also into the endothelial lineage. We isolated and cloned a population of breast tumour stem cells, cultured as mammospheres that expressed the stem markers nestin and Oct-4 and not epithelial and endothelial differentiation markers, and formed serially transplantable tumours in SCID mice. When cultured in the presence of serum, mammosphere-derived clones differentiated in the epithelial lineage. When cultured in the presence of VEGF, the same clones were also able to differentiate in the endothelial lineage acquiring endothelial markers and properties, such as the ability to organize in Matrigel into capillary-like structures. In the transplanted tumours, originated from mammospheres, we demonstrate that some of the intratumour vessels were of human origin, suggesting an in vivo endothelial differentiation of mammosphere-derived cells. Finally, endothelial cell clones originated from mammospheres were able, when implanted in Matrigel in SCID mice, to form after 7 days a human vessel network and, after 3-4 weeks, an epithelial tumour suggesting that in the endothelial-differentiated cells a tumourigenic stem cell population is maintained. In conclusion, the results of the present study demonstrate that stem cells of breast cancer have the ability to differentiate not only in epithelial but also in endothelial lineage, further supporting the hypothesis that the tumour-initiating population possesses stem cell characteristics relevant for tumour growth and vascularization. PMID- 18410531 TI - Direct and indirect fitness consequences of female choice in a crustacean. AB - Understanding the evolution and maintenance of female mate choice requires information on both the benefits (the sum of direct and indirect benefits) and costs of selective mating. In this study, I assessed the fitness consequences of female mate choice in a freshwater crustacean. In Hyalella amphipods, males attempt to form precopulatory pairs with females. Large males, bearing large posterior gnathopods, tend to be over-represented in precopulatory pairs. I show that females receive both direct (reduced risk of predation while paired) and indirect (sexy sons) benefits from mating with these males. Furthermore, the behavioral mechanisms used to filter male phenotypes carry no detectable energetic cost for females. Thus, females that choose males with successful phenotypes are expected to have higher Darwinian fitness than females that mate at random. This study shows that direct and indirect selection act together to favor large male size, which explains the sexual size dimorphism and size-based mating biases observed in this species. PMID- 18410529 TI - Resistin modulates glucose uptake and glucose transporter-1 (GLUT-1) expression in trophoblast cells. AB - The adipocytokine resistin impairs glucose tolerance and insulin sensitivity. Here, we examine the effect of resistin on glucose uptake in human trophoblast cells and we demonstrate that transplacental glucose transport is mediated by glucose transporter (GLUT)-1. Furthermore, we evaluate the type of signal transduction induced by resistin in GLUT-1 regulation. BeWo choriocarcinoma cells and primary cytotrophoblast cells were cultured with increasing resistin concentrations for 24 hrs. The main outcome measures include glucose transport assay using [(3)H]-2-deoxy glucose, GLUT-1 protein expression by Western blot analysis and GLUT-1 mRNA detection by quantitative real-time RT-PCR. Quantitative determination of phospho(p)-ERK1/2 in cell lysates was performed by an Enzyme Immunometric Assay and Western blot analysis. Our data demonstrate a direct effect of resistin on normal cytotrophoblastic and on BeWo cells: resistin modulates glucose uptake, GLUT-1 messenger ribonucleic acid (mRNA) and protein expression in placental cells. We suggest that ERK1/2 phosphorylation is involved in the GLUT-1 regulation induced by resistin. In conclusion, resistin causes activation of both the ERK1 and 2 pathway in trophoblast cells. ERK1 and 2 activation stimulated GLUT-1 synthesis and resulted in increase of placental glucose uptake. High resistin levels (50-100 ng/ml) seem able to affect glucose uptake, presumably by decreasing the cell surface glucose transporter. PMID- 18410530 TI - Progressive silencing of p14ARF in oesophageal adenocarcinoma. AB - The frequency of oesophageal adenocarcinoma is increasing in Western countries for unknown reasons, and correlates with a corresponding increase in the pre malignant condition, Barrett's Oesophagus, which raises the risk of adenocarcinoma by some 40- to 125-fold. We have examined how disease progression correlates with changes in expression of the p14ARF (ARF) tumour suppressor, a key regulator of the p53 tumour suppressor pathway that is silenced in some 30% of cancers overall, but for which a role in oesophageal cancer is unclear. We have used quantitative PCR, RT-PCR, methylation-specific PCR and chromatin immunoprecipitation to examine the regulation and function of ARF in oesophageal adenocarcinoma tissue specimens and cell lines. We find highly significant reductions (P< 0.001) in ARF expression during disease progression from normal oesophageal epithelium to Barrett's Oesophagus to adenocarcinoma, with 57/76 (75%) adenocarcinomas displaying undetectable levels of ARF expression. Retention of ARF expression in adenocarcinoma is a highly significant indicator of increased survival (P< 0.001) and outperforms all clinical variables in a multivariate model. CpG methylation as well as histone H3 methylation of lysines 9 and 27 contribute independently to ARF gene silencing in adenocarcinoma cell lines and can be reversed by 5-aza-2'-deoxycytidine. The results suggest that silencing of ARF is involved in the pathogenesis of oesophageal adenocarcinoma and show that either DNA or histone methylation can provide the primary mechanism for ARF gene silencing. Silencing of ARF could provide a useful marker for increased risk of progression and poor prognosis. PMID- 18410532 TI - Sexual selection and interacting phenotypes in experimental evolution: a study of Drosophila pseudoobscura mating behavior. AB - Sexual selection requires social interactions, particularly between the sexes. When trait expression is influenced by social interactions, such traits are called interacting phenotypes and only recently have the evolutionary consequences of interacting phenotypes been considered. Here we investigated how variation in relative fitness, or the opportunity for sexual selection, affected the evolutionary trajectories of interacting phenotypes. We used experimentally evolved populations of the naturally promiscuous Drosophila pseudoobscura, in which the numbers of potential interactions between the sexes, and therefore relative fitness, were manipulated by altering natural levels of female promiscuity. We considered two different mating interactions between the sexes: mating speed and copulation duration. We investigated the evolutionary trajectories of means and (co)variances (P) and also the influence of genetic drift on the evolutionary response of these interactions. Our sexual selection treatments did not affect the means of either mating speed or copulation duration, but they did affect P. We found that the means of both traits differed among replicates within each selection treatment whereas the Ps did not. Changes as a consequence of genetic drift were excluded. Our results show that although variable potential strengths of sexual interactions influence the evolution of interacting phenotypes, the influence may be nonlinear. PMID- 18410533 TI - A rare model limits the distribution of its more common mimic: a twist on frequency-dependent Batesian mimicry. AB - Batesian mimics are predicted to lose their fitness advantage not only in the absence of an unpalatable model, but also when the mimic becomes relatively abundant. The phenotypic hybrid zone between mimetic and nonmimetic admiral butterflies, comprising the polytypic Limenitis arthemis species complex, offers an ideal opportunity to test these predictions because the position of the hybrid zone is hypothesized to be controlled by the geographic range of Battus philenor, the chemically defended model. We used 29 years of observational field data from a continental-scale butterfly monitoring program, the 4th of July Butterfly Counts, to show that (1) the advantage of mimicry does not extend beyond the range of the model, (2) in contrast to expectations, the mimicry complex is maintained even where the model is rare and (3) the sharp phenotypic transition between mimetic and nonmimetic admiral populations occurs over a very narrow spatial scale corresponding to the limit of the model's range. These results suggest that, even at very low densities, there is selection for Batesian mimicry and it maintains the geographic position of this hybrid zone. Our findings highlight the value of large-scale, long-term citizen science monitoring programs for answering basic ecological and evolutionary questions. PMID- 18410534 TI - Heritability and correlation structure of nectar and floral morphology traits in Nicotiana alata. AB - The heritability and genetic basis of nectar traits have been rarely studied in the field, where plants are exposed to environmental factors that could mask underlying genetic effects. Heritabilities and variance components were estimated for nectar and morphological traits of Nicotiana alata, using a partial diallel design. The main experiment was conducted in a Missouri experimental garden using a randomized block design with three plant density treatments, whereas a smaller experiment was conducted near native Brazil habitat to compare the environmental variance in traits between Missouri and Brazil. Significant heritability was detected for nectar volume and energy content, and for corolla tube length. Phenotypic correlations were significant between all traits investigated, whereas significant genetic correlations were only found between nectar volume and energy and between corolla limb width and mouth diameter. There were no significant family-by-density interactions detected in the Missouri field environment. All traits differed significantly between Missouri and Brazil environments, but significant genetic by environment (G x E) interactions between Missouri and Brazil were detected for only one trait. This study shows that nectar traits can be heritable despite considerable environmental variation. PMID- 18410535 TI - Did tectonic activity stimulate oligo-miocene speciation in the Indo-West Pacific? AB - Analyses of molecular phylogenies of three unrelated tropical marine gastropod genera, Turbo, Echinolittorina, and Conus, reveal an increase in the rate of cladogenesis of some Indo-West Pacific (IWP) clades beginning in the Late Oligocene or Early Miocene between 23.7 and 21.0 million years ago. In all three genera, clades with an increased rate of diversification reach a maximum of diversity, in terms of species richness, in the central IWP. Congruence in both the geographical location and the narrow interval of timing suggests a common cause. The collision of the Australia and New Guinea plate with the southeast extremity of the Eurasian plate approximately 25 Mya resulted in geological changes to the central IWP, including an increase in shallow-water areas and length of coastline, and the creation of a mosaic of distinct habitats. This was followed by a period of rapid diversification of zooxanthellate corals between 20 and 25 Mya. The findings reported here provide the first molecular evidence from multiple groups that part of the present-day diversity of shallow-water gastropods in the IWP arose from a rapid pulse of speciation when new habitats became available in the Late Oligocene to Early Miocene. After the new habitats were filled, the rate of speciation likely decreased and this combined with high levels of extinction (in some groups), resulted in a slow down in the rate of diversification in the genera examined. PMID- 18410536 TI - Is floral specialization an evolutionary dead-end? Pollination system transitions in Ruellia (Acanthaceae). AB - Pollination systems frequently reflect adaptations to particular groups of pollinators. Such systems are indicative of evolutionary specialization and have been important in angiosperm diversification. We studied the evolution of pollination systems in the large genus Ruellia. Phylogenetic analyses, morphological ordinations, ancestral state reconstructions, and a character mapping simulation were conducted to reveal key patterns in the direction and lability of floral characters associated with pollination. We found significant floral morphological differences among species that were generally associated with different groups of floral visitors. Floral evolution has been highly labile and also directional. Some specialized systems such as hawkmoth or bat pollination are likely evolutionary dead-ends. In contrast, specialized pollination by hummingbirds is clearly not a dead-end. We found evidence for multiple reverse transitions from presumed ancestral hummingbird pollination to more derived bee or insect pollination. These repeated origins of insect pollination from hummingbird-pollinated ancestors have not evolved without historical baggage. Flowers of insect-pollinated species derived from hummingbird pollinated ancestors are morphologically more similar to hummingbird flowers than they are to other more distantly related insect-pollinated flowers. Finally, some pollinator switches were concomitant with changes in floral morphology that are associated with those pollinators. These observations are consistent with the hypothesis that some transitions have been adaptive in the evolution of Ruellia. PMID- 18410537 TI - Defined order of evolutionary adaptations: experimental evidence. AB - Organisms often adapt to new conditions by means of beneficial mutations that become fixed in the population. Often, full adaptation requires several different mutations in the same cell, each of which may affect a different aspect of the behavior. Can one predict order in which these mutations become fixed? To address this, we experimentally studied evolution of Escherichia coli in a growth medium in which the effects of different adaptations can be easily classified as affecting growth rate or the lag-phase duration. We find that adaptations are fixed in a defined and reproducible order: first reduction of lag phase, and then an increase of the exponential growth rate. A population genetics theory explains this order, and suggests growth conditions in which the order of adaptations is reversed. We experimentally find this order reversal under the predicted conditions. This study supports a view in which the evolutionary path to adaptation in a new environment can be captured by theory and experiment. PMID- 18410538 TI - Fixation probabilities depend on life history: fecundity, generation time and survival in a burst-death model. AB - The burst-death model has been developed to describe the life history of organisms with variable generation times and a burst of a fixed number of offspring. The model also includes an optional constant clearance rate, such as washout from a chemostat, and the possibility of sustained periods of population growth followed by severe bottlenecks, as in serial passaging. In this model, a beneficial mutation can either increase the burst rate or the burst size, or reduce the clearance rate, thus increasing survival. In this article we examine the effects of these three possible mechanisms on both the Malthusian fitness and the fixation probability of the lineage. We find that equivalent relative increases in the burst rate or burst size confer equivalent increases in the Malthusian fitness of a lineage, whereas increasing survival typically has a more moderate effect on Malthusian fitness. In contrast, for beneficial mutations that confer the same increase in fitness, mutations that increase survival are the most likely to fix, followed by mutations that increase the burst rate. Mutations that increase the burst size are the least likely to fix. These results imply that mutant lineages with the highest Malthusian fitness are not, in many cases, the most likely to escape extinction. PMID- 18410539 TI - Role of type IV secretion in Helicobacter pylori pathogenesis. AB - Helicobacter pylori is a highly successful human-specific gastric pathogen that colonizes more than half the world's population. Infection with this bacterium can induce gastric pathologies ranging from chronic gastritis to peptic ulcers and even cancer. Virulent H. pylori isolates harbour the cag (cytotoxin associated genes) pathogenicity island, a 40 kb stretch of DNA that encodes components of a type IV secretion system (T4SS). This T4SS forms a pilus for the injection of virulence factors into host target cells such as the CagA oncoprotein. This is accomplished by a specialized adhesin of the pilus surface, the CagL protein, which binds to and activates host cell integrins for subsequent delivery of CagA across the host cell membrane. Injected CagA becomes tyrosine phosphorylated by Src and Abl family kinases and mimics a host cell protein in binding and activation of multiple signalling factors. Here we review the recent advances in the characterization of phosphorylation-dependent and phosphorylation independent signalling activities of CagA and the T4SS which include the induction of membrane dynamics, actin cytoskeletal rearrangements and the disruption of cell-to-cell junctions as well as proliferative, pro-inflammatory and anti-apoptotic nuclear responses. The contribution of these signalling cascades to H. pylori pathogenesis is discussed. PMID- 18410540 TI - Sterile abscesses complicating monoclonal gammopathy of undetermined significance. PMID- 18410541 TI - Gene expression profiling of Polycomb, Hox and Meis genes in patients with acute myeloid leukaemia. AB - The Polycomb group (PcG) of genes is important for differentiation and cell-cycle regulation and is aberrantly expressed in several cancers. To analyse the role of deregulated PcG genes in acute myeloid leukaemia (AML), we determined by RQ-PCR the expression of the PcG genes BMI-1, MEL18, SCML2, YY1 and EZH2, and the downstream PcG targets HOXA4, HOXA9 and MEIS1 in diagnostic bone marrow samples from 126 AML patients. There was a general overexpression of the genes in AML patients compared to 20 healthy donors, except of HOXA4 and MEL18, which both displayed a wide range of expression levels within the AML subgroups. Among the AML patients with normal karyotype, a low HOXA4 level was associated with a shorter overall survival (P = 0.005). In addition, expression levels of MEL18 and EZH2 were significantly (P < 0.025) higher in patients with complex karyotype and lower in CBF-mutated patients. The t(8;21) vs. inv(16) positive patients showed significantly different expression of SCML2, BMI-1, YY1, HOXA9 and MEIS1 (P < or = 0.01). Comparisons between the PcG and PcG-regulated genes and a number of clinical and molecular data revealed correlations to genes involved in DNA methylation (DNMT1, DNMT3B), apoptosis (BAX, CASPASE 3) and multidrug-resistance (MDR1, MRP ) (P < 0.01). In conclusion, our data suggest that the role of PcG and PcG-regulated genes in leukaemogenesis varies between, as well as within karyotypic subgroups. PMID- 18410542 TI - Stem cell transplantation in poor-risk chronic lymphocytic leukemia: assessment of post-transplant minimal residual disease using four- and six-color flow cytometry and allele-specific RQ-PCR. AB - A total of 178 bone marrow samples were taken for minimal residual disease (MRD) analysis after 34 stem cell transplantations for poor-risk chronic lymphocytic leukemia, and 86 of them were analyzed in parallel by flow cytometry and allele specific oligonucleotide-PCR (ASO-PCR). ASO primer was successfully designed for all patients whose frozen diagnosis samples were available. Flow cytometry and ASO-PCR were concordant, i.e. both either positive or both negative, in 78% of the analyses. Flow cytometry did not detect MRD in any of the samples that were PCR-negative cases. In contrast, ASO-PCR detected MRD in samples that were negative for MRD by flow cytometry in 22% of the analyses. In one patient, the immunophenotype but not the IgV(H) gene sequence had changed during a course of the disease, and MRD could not be followed by flow cytometry. In the remaining cases, the discrepancy was due to a higher sensitivity of ASO-PCR. Autologous stem cell transplantation resulted in clinical complete response in 87% (20/23) of the patients. By flow cytometry, 35% (8/23) of autotransplanted patients became MRD-negative, but only 12.5% (2/16) PCR-negative (sensitivity of ASO-PCR <0.001 and <0.01, respectively). All allotransplanted patients achieved or maintained hematological CR, and five out of nine patients (56%) became PCR negative (sensitivity of PCR between <0.001 and <0.003), two of them having non myeloablative conditioning. None of the patients who became PCR-negative after allogeneic transplantation have relapsed. PMID- 18410543 TI - Primary aldosteronism (PA) and endothelial progenitor cell (EPC) bioavailability. AB - OBJECTIVE: Patients with primary aldosteronism (PA) experience more cardiovascular events than patients with essential hypertension matched for risk factor profile. Endothelial progenitor cells (EPC) represent a bone marrow derived cell population implicated in vascular healing whose number correlates to the cardiovascular risk factor profile. Aldosterone has been reported to decrease EPC proliferation in rats. DESIGN AND PATIENTS: We assessed (i) the growth characteristics of EPC from six PA patients and six matched normotensive controls; (ii) the growth characteristics of EPC treated with increasing doses of aldosterone. MEASUREMENTS: Senescence and cell-cycle analysis of EPC from PA patients and normotensive controls and of aldosterone-treated EPC from healthy volunteers. RESULTS: No difference was found in the senescence rate between EPC from PA patients (72.4% senescent cells) and controls (70.7%, P > 0.05). No difference was also found in the cell-cycle distribution determined by FACS (controls: 75.2% cells in G0/G1 phase; PA: 73.5%, P > 0.05). Incubation of EPC with aldosterone did not modify their senescence rate (controls: 72.4% senescent cells; aldosterone 10 nmol/l: 70.9%; aldosterone 100 nmol/l 71.6%, P > 0.05 for all comparisons) and cell-cycle distribution (controls: 73.3% cells in G0/G1 phase; aldosterone 10 nmol/l: 74.9%; aldosterone 100 nmol/l: 75.4%, P > 0.05 for all comparisons). No expression of the mineralocorticoid receptor (MR) transcript was found in EPC by RT-PCR analysis. CONCLUSIONS: High aldosterone levels, both in PA patients and in vitro, exert no direct or indirect effect on EPC growth characteristics. PMID- 18410544 TI - Long-term basal and dynamic evaluation of hypothalamic-pituitary-adrenal (HPA) axis in acromegalic patients. AB - OBJECTIVE: Long-term effects of trans-naso-sphenoidal surgery (TNS) or long acting somatostatin analogs (SSA) on the function of hypothalamic-pituitary adrenal (HPA) axis have been poorly investigated. Aim of this study was to evaluate HPA axis integrity during the follow-up in patients with GH-secreting pituitary adenomas and preserved HPA function post-TNS or prior SSA. DESIGN AND PATIENTS: This retrospective study investigated 36 acromegalic patients (16M and 20F, age: 47 +/- 13 years), 20 of whom cured by TNS and 16 controlled by SSA therapy (12 previously operated and 4 in primary medical therapy), before and after long-term follow-up (median: 72 months, range: 12-240). No patient previously underwent radiotherapy. MEASUREMENTS: HPA function was studied by morning circulating cortisol and ACTH levels, 24-h urinary free cortisol (UFC) and cortisol response to low-dose short Synacthen test (LDSST, 1 microg) with a peak > 500 nmol/l as cut-off for normal function. RESULTS: Serum basal cortisol, ACTH and UFC levels were in the normal range and did not significantly change over time. As far as the cortisol peak after LDSST is concerned, 12 patients (32%, 8 TNS and 4 SSA) developed biochemical hypoadrenalism. None of the patients in primary medical therapy showed cortisol peak < 500 nmol/l. No significant correlations between HPA axis deterioration and follow-up duration, serum GH/IGF I levels, occurrence of other pituitary deficiencies, presence of secondary empty sella, changes in tumour or residual volume were observed. CONCLUSIONS: The HPA axis function must be carefully monitored over the time by dynamic testing in all acromegalic patients, independently from the type of treatment. PMID- 18410545 TI - IGF system and peripheral arterial disease: relationship with disease severity and inflammatory status of the affected limb. AB - OBJECTIVES: IGF-1 and its binding proteins are involved in the pathogenesis of atherosclerosis. We designed this study to unravel the relationship of the IGF system with peripheral arterial disease (PAD). DESIGN: Case-control, cross sectional study. MEASUREMENTS: Serum levels of IGF-1, IGFBP-3 and acid labile subunit (ALS) were measured in 96 PAD patients and 89 controls. In 28 patients who underwent peripheral angiography, C-reactive protein (CRP), IGF-1, IGFBP-3 and ALS were measured in blood from femoral vein of the affected limb and aorta. RESULTS: Compared to controls, PAD patients showed lower levels of IGFBP-3 (3569 +/- 115 vs. 3106 +/- 107 microg/l, P < 0.01), and ALS (12.2 +/- 0.5 vs. 8.3 +/- 0.5 mg/l, P < 0.01). In PAD, concentrations of IGFBP-3 and ALS were significantly lower in patients with ankle/brachial index less than median than in those with a less severe PAD. In the affected limb, CRP venous-arterial difference correlated negatively with that of IGF-1 (rho = -0.57, P < 0.01), and positively with that of IGFBP-3 (rho = 0.63, P < 0.01). At multivariate analysis, a high transfemoral gradient of CRP was independently associated with a low transfemoral gradient of IGF-1 (beta coefficient = -0.48, P < 0.01), and a high transfemoral gradient of IGFBP-3 (beta coefficient = 0.22, P < 0.05). CONCLUSIONS: This study is the first to demonstrate that the systemic levels of IGF axis components are associated with the presence and severity of PAD, and that the inflammatory status of the ischaemic limb affects the transfemoral concentrations of IGF-1 and IGFBP-3. Due to the importance of IGF axis in modulating atherosclerotic plaque progression, our data may contribute to a better understanding of PAD pathophysiology. PMID- 18410546 TI - Diagnosis and management of amiodarone-induced thyrotoxicosis: similarities and differences between North American and European thyroidologists. AB - OBJECTIVE: To investigate how North American thyroidologists assess and treat amiodarone-induced thyrotoxicosis (AIT) and to compare the results with those of the same questionnaire-based survey previously carried out among European thyroidologists. DESIGN: Members of the American Thyroid Association (ATA) with clinical interests were sent by e-mail a questionnaire on the diagnosis and management of AIT, 115 responses were received from the United States and Canada, representing about one-third of ATA members with clinical interests. RESULTS: The majority of respondents (91%vs. 68% in Europe, P < 0.05) see < 10 new cases of AIT per year, and AIT seems less frequent than amiodarone-induced hypothyroidism (AIH) in North America (34% and 66% of amiodarone-induced thyroid dysfunction, respectively, vs. 75% and 25%, respectively, in Europe, P < 0.001). When AIT is suspected, in North America hormonal assessment is mostly based on serum free T4 (FT4) and TSH measurements, while serum free T3 (FT3) determination is requested less frequently than in Europe; thyroid autoimmunity is included in the initial assessment less than in Europe. Most commonly used additional diagnostic procedures include, as in Europe, thyroid colour-flow Doppler sonography, and to a lesser extent, thyroid radioactive iodine uptake and scan, but Europeans tend to request multiple tests more than North Americans. Withdrawal of amiodarone is more often considered unnecessary by North American thyroidologists (21%vs. 10% in Europe in type 1 AIT, P < 0.05, 34%vs. 20% in type 2 AIT, P < 0.05). In type 1 AIT thionamides represent the treatment of choice for North Americans as well as for Europeans, but the former use them as monotherapy in 65%vs. 51% of Europeans (P < 0.05) who more often consider potassium perchlorate as an useful addition (31%vs. 15% of North Americans, P < 0.01). Glucocorticoids are the selected treatment for type 2 AIT, alone (62%vs. 46% in Europe, P < 0.05) or in association with thionamides (16%vs. 25% in Europe, P = NS). After restoration of euthyroidism, thyroid ablation in the absence of recurrent thyrotoxicosis is recommended in type 1 AIT less frequently by North Americans. If amiodarone therapy needs to be reinstituted, prophylactic thyroid ablation is advised by 76% in type 1 AIT, while a 'wait-and-see' strategy is adopted by 61% in type 2 AIT, similar to behaviour of European thyroidologists. CONCLUSION: Similarities and differences exist between expert North American and European thyroidologists concerning the diagnosis and management of AIT. While differences reflect the frequent uncertainty of the underlying mechanism leading to AIT, similarities may represent the basis to refine the diagnostic criteria and to improve the therapeutic outcomes of this challenging clinical situation. PMID- 18410547 TI - Polymorphisms in the brain-specific thyroid hormone transporter OATP1C1 are associated with fatigue and depression in hypothyroid patients. AB - INTRODUCTION: Some hypothyroid patients continue to have significant impairments in psychological well-being, despite adequate treatment with levothyroxine (LT4). T4 transport across the blood-brain barrier is one of the crucial processes for thyroid hormone action in the brain. OATP1C1, a thyroid hormone transporter expressed at the blood-brain barrier, is considered to play a key role in delivering serum T4 to the brain. OBJECTIVE: To examine whether polymorphisms in OATP1C1 are determinants of well-being, neurocognitive functioning and preference for replacement therapy with a combination of LT4 and liothyronine (LT3). DESIGN AND PARTICIPANTS: We studied 141 patients with primary autoimmune hypothyroidism, adequately treated with LT4 monotherapy and participating in a randomized clinical trial comparing LT4 therapy with LT4-LT3 combination therapy. OUTCOME MEASUREMENTS: Different questionnaires on well-being and neurocognitive tests were performed at baseline. Serum thyroid parameters, OATP1C1-intron3C > T, OATP1C1-Pro143Thr and OATP1C1-C3035T polymorphisms were determined. RESULTS: Allele frequencies of the OATP1C1 polymorphisms in patients with primary hypothyroidism were similar to those of healthy controls. Both the OATP1C1 intron3C > T and the OATP1C1-C3035T polymorphism, but not the OATP1C1-Pro143Thr polymorphism, were associated with symptoms of fatigue and depression. OATP1C1 polymorphisms were not associated with measures of neurocognitive functioning or preference for combined LT4-LT3 therapy. CONCLUSIONS: OATP1C1 polymorphisms are associated with fatigue and depression, but do not explain differences in neurocognitive functioning or appreciation of LT4-LT3 combination therapy. Future studies are needed to confirm these findings. PMID- 18410548 TI - Aryl hydrocarbon receptor interacting protein (AIP) gene mutation analysis in children and adolescents with sporadic pituitary adenomas. AB - OBJECTIVE: Pituitary adenomas occur rarely in childhood and adolescence. Pituitary adenoma predisposition (PAP) has been recently associated with germline mutations in the aryl hydrocarbon receptor interacting protein (AIP) gene. The aim of the study was to examine the proportion of germline AIP mutations in apparently sporadic paediatric pituitary adenomas. DESIGN: Genomic DNA was analysed for mutations in the AIP gene, by PCR amplification and direct sequencing. PATIENTS: A population-based cohort consisting of 36 apparently sporadic paediatric pituitary adenoma patients, referred to two medical centres in Italy, was included in the study. Patients were either less than 18 years at diagnosis, or showed clinical evidence of adenoma development before the age of 18 years. RESULTS: A heterozygous in-frame deletion Y248del (c.742_744delTAC) was identified in one GH-secreting adenoma patient. Loss of heterozygosity (LOH) analysis of tumour DNA revealed the loss of the wild-type allele. First degree relatives carrying the mutation were clinically unaffected. CONCLUSIONS: While mutations were absent in non-GH-secreting adenoma patients, germline AIP mutations can be found in children and adolescents with GH-secreting tumours, even in the absence of family history. The present study reports the AIP mutation analysis results on patients of a single ethnic origin. Clearly, further studies are needed to improve our knowledge on the role of AIP in paediatric pituitary adenomas. PMID- 18410549 TI - Absence of sonic hedgehog (Shh) germline mutations in patients with thyroid dysgenesis. PMID- 18410550 TI - Correlation of circulating full-length visfatin (PBEF/NAMPT) with metabolic parameters in subjects with and without diabetes: a cross-sectional study. AB - OBJECTIVE: Here we use a novel ELISA that is specific for full-length visfatin (PBEF/NAMPT), compare it with the existing C-terminal based assay and use it to investigate associations of visfatin with metabolic parameters. DESIGN, PATIENTS AND MEASUREMENTS: We established the specificity and effectiveness of the new ELISA and evaluated the associations of full-length visfatin with clinical, anthropometric and metabolic parameters in a cross-sectional study of 129 Thai subjects, consisting of 50 outpatients with type 2 diabetes and 79 healthy volunteers. RESULTS: The new ELISA accurately recovered full-length recombinant visfatin and detected visfatin secreted by primary human and rat adipocytes. We found serum full-length visfatin was significantly higher in subjects with diabetes compared to their nondiabetic peers (median 2.75 vs. 2.22 ng/ml, P = 0.0142). After adjustment for age, gender and traditional metabolic risk factors, adjusted mean visfatin remained significantly higher in the diabetes group (3.80 vs. 2.10 ng/ml, P = 0.0021). On Spearman univariate correlation analysis, visfatin was significantly associated with resistin (r = 0.30, P = 0.0011), but not with any other anthropometric or metabolic variables, including adiponectin multimers. On multiple linear regression analysis, the only covariates independently associated with visfatin were diabetes (t = 3.11, P = 0.0024) and log resistin (t = 2.68, P = 0.0086). CONCLUSIONS: Circulating visfatin is independently associated with diabetes and resistin concentration, but is not related to adiponectin multimers or other metabolic covariates. These data are suggestive of a potential role of visfatin in subclinical inflammatory states. PMID- 18410551 TI - Changes in endothelial function and its association with plasma osteoprotegerin in hypothyroidism with exercise-induced silent myocardial ischaemia. AB - OBJECTIVE: Hypothyroidism is associated with an increased risk for cardiovascular disease. Exercise-induced silent myocardial ischaemia (SI) is an early stage of coronary artery disease. Recently, many studies have shown that endothelial dysfunction is an early physiological event in atherosclerosis, and osteoprotegerin (OPG) acts as an important regulatory molecule in the vasculature. The aim of this study was to investigate the alteration of endothelial function and its association with plasma OPG in hypothyroidism with SI. METHODS: Forty-eight female postmenopausal hypothyroid patients with normal rest electrocardiography (ECG) were selected. Of these, 19 cases had SI. Twenty healthy females without SI were selected as controls. High-resolution ultrasound was used to measure brachial artery diameter at rest, after reactive hyperaemia and after sublingual glyceryltrinitrate (GTN). Plasma OPG concentration was measured in duplicate by a sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: Flow-mediated arterial dilation (FMD) in the total hypothyroid group, the hypothyroidism with SI group and the hypothyroidism without SI group was 3.51 +/- 0.62%, 3.20 +/- 0.54% and 3.72 +/- 0.60%, respectively, significantly lower than that in the controls (5.08 +/- 0.61%) (P < 0.01). Compared with the hypothyroidism without SI group, FMD in the hypothyroidism with SI group was significantly lower (P < 0.05). Plasma OPG levels in the total hypothyroid group, patients with SI and patients without SI were significantly higher than in the control group (P < 0.05). Compared with patients without SI, OPG levels were significantly higher in patients with SI (P < 0.05). On multiple regression analysis, low density lipoprotein cholesterol (LDL-C), lipoprotein (a) [Lp(a)], C reactive protein (CRP), OPG, TSH, free T3 (FT3) and thyroid peroxidase antibody (TPO-Ab) were found to be significant factors that were associated with FMD. Logistic analysis also showed that LDL-C, TSH, OPG, CRP and FMD were independently and significantly associated with SI in hypothyroidism. CONCLUSION: Impaired endothelial function and increased levels of OPG exist in hypothyroid patients, especially those with SI. These findings support the growing concept that endothelial dysfunction may be associated with vascular disease, and subsequently elevated plasma OPG may have a role in the development of vascular dysfunction in hypothyroid patients. PMID- 18410552 TI - Depot-specific steroidogenic gene transcription in human adipose tissue. AB - CONTEXT: Sex steroids (androgens and oestrogens) and corticosteroids (glucocorticoids and mineralocorticoids) have a major impact on fat distribution. Several genes involved in steroid synthesis and metabolism, such as 11beta hydroxysteroid dehydrogenase type 1 and aromatase, are known to be expressed within adipose tissue, thus modulating local steroid levels; however, our knowledge of which genes are expressed and at what level is incomplete. OBJECTIVE: To detect by real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) which of 13 key steroidogenic genes are transcribed within human adipose tissue and to assess whether mRNA levels differ significantly between the subcutaneous abdominal and omental adipose depots. PATIENTS: Eight women undergoing caesarean section [age 29.1 +/- 6.5 years, body mass index (BMI) 28.9 +/- 8.4 kg/m(2)]. RESULTS: Genes transcribed in both depots were StAR (steroidogenic acute regulatory protein), CYP11A1 (side-chain cleavage enzyme), HSD3B2 (3beta-hydroxysteroid dehydrogenase type 2), CYP21B (21 hydroxylase), CYP19 (aromatase), HSD11B1 (11beta-hydroxysteroid dehydrogenase type 1), HSD17B3, HSD17B5, HSD17B7 (17beta-hydroxysteroid dehydrogenase types 3, 5 and 7) and SRD5A2 (5alpha-reductase type 2). All but SRD5A2 varied significantly in abundance between depots. CYP17 (17alpha-hydroxylase), CYP11B1 (11beta-hydroxylase) and CYP11B2 (aldosterone synthase) transcription were not detected. CONCLUSIONS: This study confirms and significantly extends our knowledge of steroidogenic gene expression within adipose tissue, showing that transcript levels are depot specific. We have demonstrated that de novo synthesis from cholesterol of sex steroids, cortisol and aldosterone is not possible because of the absence of key steroidogenic mRNAs. Instead, the pattern of transcription suggests that 11-deoxycorticosterone, a mineralocorticoid, would be the ultimate product of any de novo adipose synthesis. PMID- 18410553 TI - A comparison between rimonabant and metformin in reducing biochemical hyperandrogenaemia and insulin resistance in patients with polycystic ovary syndrome (PCOS): a randomized open-label parallel study. AB - CONTEXT: Weight loss and metformin therapy are reported to be beneficial in improving the biochemical hyperandrogenaemia and insulin resistance of polycystic ovary syndrome (PCOS). Rimonabant has been found to reduce weight and improve the metabolic profile in patients with obesity, type 2 diabetes and metabolic syndrome. OBJECTIVE: To compare the effects of insulin sensitization with metformin to weight reduction by rimonabant on biochemical hyperandrogenaemia and insulin resistance in patients with PCOS. DESIGN: A randomized, open-label parallel study. SETTING: Endocrinology outpatient clinic in a referral centre. SUBJECTS: Twenty patients with PCOS and biochemical hyperandrogenaemia with a body mass index (BMI) >or= 30 kg/m(2) were recruited. INTERVENTION: Patients were randomized to 1.5 g daily of metformin or 20 mg daily of rimonabant. MAIN OUTCOME MEASURES: The primary end-point of the study was a change in total testosterone. RESULTS: After 12 weeks of rimonabant there was a significant reduction (mean +/- SEM) in weight (104.6 +/- 4.6 vs. 98.4 +/- 4.7 kg, P < 0.01), waist circumference (116.0 +/- 3.3 vs. 109.2 +/- 3.7 cm, P < 0.01), hip circumference (128.5 +/- 4.0 vs. 124.1 +/- 4.2 cm, P < 0.03), waist-hip ratio (0.90 +/- 0.02 vs. 0.88 +/- 0.01, P < 0.01) free androgen index (FAI) (26.6 +/- 6.1 vs. 16.6 +/- 4.1, P < 0.01), testosterone [4.6 +/- 0.4 vs. 3.1 +/- 0.3 nmol/l (132.7 +/- 11.5 vs. 89.4 +/- 8.65 ng/dl), P < 0.01] and insulin resistance as measured by the homeostasis model assessment (HOMA) method (4.4 +/- 0.5 vs. 3.4 +/- 0.4, P = 0.05). There was no change in any of these parameters in the metformin-treated group. CONCLUSION: This study suggests that the weight loss through rimonabant therapy may be of use in patients with PCOS and appears superior to insulin sensitization by metformin in reducing the FAI and insulin resistance in obese PCOS patients treated over a 12-week period. PMID- 18410554 TI - Discriminative power of three indices of renal calcium excretion for the distinction between familial hypocalciuric hypercalcaemia and primary hyperparathyroidism: a follow-up study on methods. AB - BACKGROUND: Familial hypocalciuric hypercalcaemia (FHH) must be differentiated from primary hyperparathyroidism (PHPT) because prognosis and treatment differ. In daily practice this discrimination is often based on the renal calcium excretion or the calcium/creatinine clearance ratio (CCCR). However, the diagnostic performance of these variables is poorly documented. AIM: To appraise the power of various simple biochemical variables to differentiate between FHH and PHPT using calcium sensing receptor (CASR) gene analysis and histopathological findings as gold standards. DESIGN: Follow-up approach (direct design). MATERIALS: We included 54 FHH patients (17 males and 37 females, aged 18 75 years) with clinically significant mutations in the CASR gene and 97 hypercalcaemic patients with histologically verified PHPT (17 males and 80 females, aged 19-86 years). All PHPT patients became normocalcaemic following successful neck exploration. RESULTS: Based on receiver operating characteristic (ROC) curve analysis, the CCCR was only marginally better, as judged by the area under curve (AUC = 0.923 +/- 0.021 (SE)), than the 24-h calcium/creatinine excretion ratio (AUC = 0.903 +/- 0.027) and the 24-h calcium excretion (AUC = 0.876 +/- 0.029). However, overlap performance analysis disclosed that the CCCR included fewer patients with PHPT together with the FHH patients than the other two variables at different cut-off points. Based on the ROC curve, the optimal cut-off point for diagnosing FHH using CCCR was < 0.0115, which yielded a diagnostic specificity of 0.88 and a sensitivity of 0.80. Overlap analysis revealed that a cut-off point for CCCR at < 0.020 would sample 98% (53/54) of all patients with FHH and include 35% (34/97) of the PHPT patients. CONCLUSION: Our results support the use of the CCCR as an initial screening test for FHH. We suggest a two-step diagnostic procedure, where the first step is based on the CCCR with a cut-off at < 0.020, and the second step is CASR gene analysis in patients with FHH or PHPT. PMID- 18410555 TI - GH and IGF-I excess control contributes to blood pressure control: results of an observational, retrospective, multicentre study in 105 hypertensive acromegalic patients on hypertensive treatment. AB - CONTEXT: Approximately one-third of patients with acromegaly have concomitant hypertension. The outcome of hypertension after treatment of acromegaly is unknown. OBJECTIVE: To evaluate the role of GH and IGF-I control on systolic (SBP) and diastolic blood pressure (DBP) levels. PATIENTS: One hundred and five hypertensive patients (60 women, 45 men) with active disease receiving treatment for hypertension at their diagnosis of acromegaly. DESIGN: Observational, retrospective, multicentre. MEASUREMENTS: At diagnosis and after 24 months (median) of treatment we measured serum GH and IGF-I levels, blood pressure levels, left ventricular (LV) mass index (LVMi), early-to-late mitral flow velocity (E/A, as a measure of diastolic function) and LV ejection fraction (LVEF, as a measure of systolic function). RESULTS: At the diagnosis of acromegaly, hypertension was mild in 41.1% and severe in 58.9%. Serum GH and IGF I levels did not differ in patients with mild or severe hypertension. After 24 months of treatment, all patients had a notable decrease in both GH and IGF-I levels, and achieved significantly lower levels of DBP, heart rate and LVMi; 76 patients (71%) had achieved control of GH and IGF-I levels. Only the patients with controlled acromegaly achieved significantly lower SBP levels and significantly improved cardiac systolic and diastolic function. A higher dose of antihypertensive drugs and/or an increased number of drugs to control hypertension were significantly greater in patients with uncontrolled (32.3%) than in those with controlled acromegaly (7.8%; P = 0.004). CONCLUSION: Hypertensive patients with controlled acromegaly achieved improved control of hypertension and of cardiac diastolic and systolic function. The use of antihypertensive drugs was significantly less in patients achieving control of acromegaly. PMID- 18410556 TI - Prospective evaluation of transient elastography for the diagnosis of hepatic fibrosis in Asians: comparison with liver biopsy and aspartate transaminase platelet ratio index. AB - BACKGROUND: Transient elastography (TE) is a reliable non-invasive predictor of hepatic fibrosis, but data on TE in Asians are limited. AIM: To evaluate prospectively the accuracy of TE for diagnosis of hepatic fibrosis in Asians compared with APRI (aspartate transaminase to platelet ratio index). METHODS: One hundred and twenty consecutive patients who underwent liver biopsy were enrolled. TE (Fibroscan) was performed by two independent operators. Fibrosis was graded by two independent pathologists using the METAVIR classification. Area under receiver operating curves (AUROC) were used to evaluate the accuracy of TE and APRI in diagnosing significant fibrosis (F >or= 2) and cirrhosis (F4). RESULTS: Predominant aetiologies were hepatitis B (48%), non-alcoholic steatohepatitis (14%) and hepatitis C (8%). TE was unsuccessful in five patients (4.2%) because of small inter-costal space (three patients), obesity and ascites. There was good correlation between TE and fibrosis (r = 0.606). AUROC for diagnosis of significant fibrosis was 0.856 (95% CI 0.779-0.932) for TE and 0.673 (95% CI 0.568-0.777) for APRI. AUROC for diagnosis of cirrhosis was 0.924 (95% CI 0.857 0.990) for TE and 0.626 (95% CI 0.437-0.815) for APRI. Optimal TE value was 9.0 kPa for diagnosis of significant fibrosis and 16.0 kPa for cirrhosis with specificity/sensitivity/PPV/NPV/accuracy of 82.6%/85.2%/80.9%/86.7%/84.1% and 88.9%/82.7%/32.0%/98.8%/83.2%, respectively. CONCLUSIONS: Transient elastography is a reliable predictor of hepatic fibrosis in Asians. Failure of TE in Asians is commonly because of small inter-costal space. TE is superior to APRI for non invasive diagnosis of hepatic fibrosis and cirrhosis. PMID- 18410557 TI - Review article: current management of non-alcoholic fatty liver disease and non alcoholic steatohepatitis. AB - BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of metabolic syndrome. AIM: To assess the epidemiological impact and the current management of patients with NAFLD. METHODS: Published peer reviewed literature and abstracts concerning NAFLD and non-alcoholic steatohepatitis (NASH) were reviewed. Articles specifically related to epidemiology, diagnosis and current treatment strategies for NAFLD and NASH are summarized. RESULTS: NAFLD is strongly associated with the epidemic of obesity and type-2 diabetes mellitus, and is estimated to affect about 20-30% of the population in the US. From the spectrum of NAFLD, only patients with biopsy proven NASH (estimated prevalence in the US population is about 3-5%) have been convincingly shown to progress to cirrhosis, liver failure and hepatocellular carcinoma. The clinical manifestation of NAFLD is usually absent or subtle, with abnormal aminotransferases or incidental radiographic findings of fatty liver. The pathogenesis of NAFLD is attributed to a multi-hit process involving insulin resistance, oxidative stress, apoptotic pathways, and adipocytokines. In 2008, there is no established treatment for NAFLD. Weight loss and treatment for each component of metabolic syndrome. Nevertheless, a large number of agents are being considered in clinical trials of patients with NASH. CONCLUSIONS: Awareness of the tremendous impact of NAFLD as an important cause of chronic liver disease is increasing along with a great deal of information about its pathogenesis. Future, well-designed clinical trials that target specific pathways involved in the pathogenesis of NASH are urgently needed. PMID- 18410558 TI - Health-economic analysis: cost-effectiveness of scheduled maintenance treatment with infliximab for Crohn's disease--modelling outcomes in active luminal and fistulizing disease in adults. AB - BACKGROUND: Infliximab has been shown to be efficacious in moderate-to-severe Crohn's disease (CD). AIM: To evaluate the cost-effectiveness of scheduled maintenance treatment with infliximab in luminal and fistulizing CD patients. METHODS: Markov models were constructed to simulate the progression of adult CD patients with and without fistulae during treatment with infliximab (5 mg/kg). Transitions were estimated from published clinical trials of infliximab. Standard care, comprising immunomodulators and/or corticosteroids was used as a comparator. An average weight of 60 kg was used to estimate the dose of infliximab. The costs and outcomes were discounted at 3.5% over 5 years. The primary effectiveness measurement was quality-adjusted life years (QALYs) estimated using EQ-5D. One-way and probabilistic sensitivity analyses were performed by varying the infliximab efficacy estimates, costs and utilities. RESULTS: The incremental cost per QALY gained was pound 26,128 in luminal CD and pound 29,752 in fistulizing CD at 5 years. Results were robust and remained in the range of pound 23,752- pound 38,848 for luminal CD and pound 27,047- pound 44,206 for fistulizing CD. Patient body weight was the most important factor affecting cost-effectiveness. CONCLUSION: Eight-week scheduled maintenance treatment with infliximab is a cost-effective treatment for adult patients suffering from active luminal or fistulizing CD. PMID- 18410559 TI - Epidemiological study of gastro-oesophageal reflux disease: reflux in spouse as a risk factor. AB - BACKGROUND: Gastro-oesophageal reflux disease (GERD) is a growing health-care problem with variable distribution. AIM: To assess GERD prevalence and risk factors and their possible correlation with pathophysiology in a population-based study. METHODS: Individuals aged 18-65 years were enrolled through random cluster sampling in Tehran. Previously validated self-administered questionnaires were used. RESULTS: Of the 2500 questionnaires, 2057 were analysed (mean age: 34.8 +/- 13.0 years, 55.1% female). Frequent GERD was seen in 18.2%. Minor symptoms increased prevalence. Female gender (OR: 1.55, 95% CI: 1.01-2.41), BMI >30 kg/m(2) (OR: 1.79, 95% CI: 1.03-3.12), less education (OR: 1.52, 95% CI: 1.02 2.27), smoking (OR: 1.83, 95% CI: 1.12-2.99), NSAID use (OR: 4.23, 95% CI: 1.66 10.74) and GERD in spouse (OR: 1.82, 95% CI: 1.18-2.82) were associated with frequent GERD on multivariable analysis. GERD in first-degree relatives (OR: 1.73, 95% CI: 1.23-2.43) and asthma (OR: 4.09, 95% CI: 1.27-13.15) correlated with infrequent GERD. Minor symptoms correlated with GERD history in first-degree relatives, coffee consumption and NSAID use. Prevalence in the past 3 months was similar to that in the past 12 months (P < 0.05). CONCLUSIONS: Gastro-oesophageal reflux disease is common in Tehran. The association of 'infrequent symptoms' with GERD history in first-degree relatives and 'frequent symptoms' with GERD history in spouse may point to the presence of yet unknown precipitating environmental factors inducing GERD in a genetically susceptible host. Minor GERD symptoms seem to have independent contribution to GERD. Assessing GERD in the past 3 months predicts prevalence in the past year. PMID- 18410562 TI - Clinical trial: effectiveness of Lactobacillus rhamnosus (strains E/N, Oxy and Pen) in the prevention of antibiotic-associated diarrhoea in children. AB - BACKGROUND: Convincing evidence that probiotic administration can lower the risk of antibiotic-associated diarrhoea is limited to certain micro-organisms. AIM: To determine the efficacy of administration of Lactobacillus rhamnosus (strains E/N, Oxy and Pen) for the prevention of antibiotic-associated diarrhoea in children. METHODS: Children (aged 3 months to 14 years) with common infections were enrolled in a double-blind, randomized, placebo-controlled trial in which they received standard antibiotic treatment plus 2 x 10(10) colony forming units of a probiotic (n = 120) or a placebo (n = 120), administered orally twice daily throughout antibiotic treatment. Analyses were by intention to treat. RESULTS: Any diarrhoea (>or=3 loose or watery stools/day for >or=48 h occurring during or up to 2 weeks after the antibiotic therapy) occurred in nine (7.5%) patients in the probiotic group and in 20 (17%) patients in the placebo group (relative risk, RR 0.45, 95% confidence interval, CI 0.2-0.9). Three (2.5%) children in the probiotic group developed AAD (diarrhoea caused by Clostridium difficile or otherwise unexplained diarrhoea) compared to nine (7.5%) in the placebo group (RR 0.33, 95% CI 0.1-1.06). No adverse events were observed. CONCLUSION: Administration of L. rhamnosus (strains E/N, Oxy and Pen) to children receiving antibiotics reduced the risk of any diarrhoea, as defined in this study. PMID- 18410560 TI - Review article: molecular, pathological and therapeutic features of human enteric neuropathies. AB - BACKGROUND: Considerable information has been gathered on the functional organization of enteric neuronal circuitries regulating gastrointestinal motility. However, little is known about the neuropathophysiological mechanisms underlying gastrointestinal motor disorders. AIM: To analyse the most important pathological findings, clinical implications and therapeutic management of idiopathic enteric neuropathies. METHODS: PubMed searches were used to retrieve the literature inherent to molecular determinants, pathophysiological bases and therapeutics of gastrointestinal dysmotility, such as achalasia, gastroparesis, chronic intestinal pseudo-obstruction, Hirschsprung's disease and slow transit constipation, to unravel advances on digestive disorders resulting from enteric neuropathies. RESULTS: Current data on molecular and pathological features of enteric neuropathies indicate that degenerative and inflammatory abnormalities can compromise the morpho-functional integrity of the enteric nervous system. These alterations lead to a massive impairment in gut transit and result in severe abdominal symptoms with associated high morbidity, poor quality of life for patients and established mortality. Many pathophysiological aspects of these severe conditions remain obscure, and therefore treatment options are quite limited and often unsatisfactory. CONCLUSIONS: This review of enteric nervous system abnormalities provides a framework to better understand the pathological processes underlying gut dysmotility, to translate this knowledge into clinical management and to foster the development of targeted therapeutic strategies. PMID- 18410563 TI - Underutilization of gastroprotective strategies in aspirin users at increased risk of upper gastrointestinal complications. AB - BACKGROUND: Aspirin use is with an increased risk of upper gastrointestinal complications (UGICs). Proton pump inhibitors (PPIs) decrease the risk of UGICs among aspirin users. The distribution of risk factors for UGIC and PPI utilization among aspirin users remains uncharacterized. AIM: To determine the prevalence and predictors of PPI use in high-risk aspirin users. METHODS: Using questionnaires and administrative records, we collected information on aspirin and PPI utilization and risk factors for UGICs from a stratified random sample of subjects with established cardiovascular disease. We calculated the proportion of aspirin users with UGIC risk factors and determined the prevalence of PPI use among aspirin users with risk factors. Regression analysis was used to determine predictors of PPI use among aspirin users. RESULTS: Overall response rate was 35%, of whom 86% were regular aspirin users. Seventy-one per cent of aspirin users had at least one risk factor (in addition to cardiac disease) for the development of UGICs. Although a history of UGIC was predictive of PPI use, 44% of aspirin users with a prior history of UGICs did not receive a concomitant PPI, and only 23% of subjects with additional UGIC risk factors were prescribed a PPI. CONCLUSION: There is a high prevalence of UGIC risk factors among aspirin users, and many are not prescribed PPIs as a gastroprotective strategy. PMID- 18410564 TI - Non-oxidative glucose disposal is reduced in type 2 diabetes, but can be restored by aerobic exercise. AB - Whole-body glucose utilization consists of mitochondrial glucose oxidation and non-oxidative glycogen synthesis. We examined whether reduction of both non oxidative glucose disposal and glucose oxidation contributes to insulin resistance in type 2 diabetes. We also examined the effects of exercise on these two components. Whole-body glucose disposal rate (GDR, mg/kg/min) was evaluated in 37 type 2 diabetic (T2DM) and 17 non-diabetic (non-DM) subjects as the mean of glucose infusion rate during steady state in the euglycaemic-hyperinsulinaemic clamp study. Glucose oxidation rates were assessed by indirect calorimetry, and non-oxidative GDR was calculated by subtracting glucose oxidation rate from GDR. Intramyocellular lipid (IMCL) content of the soleus muscle was measured using (1)H-magnetic resonance spectroscopy. In 10 T2DM subjects, the changes in oxidative and non-oxidative glucose disposal during clamp were examined after 3 month exercise intervention. GDR (2.93 +/- 1.55 vs. 4.55 +/- 1.83, p = 0.001) and non-oxidative GDR (1.45 +/- 1.52 vs. 3.01 +/- 1.87, p = 0.002) were significantly lower in T2DM than in non-DM subjects. Glucose oxidation rate was comparable in the two groups, and inversely correlated with IMCL (n = 15, r =-0.565, p = 0.028). GDR (2.28 +/- 1.67 to 4.63 +/- 2.42, p = 0.021) and non-oxidative GDR (0.72 +/- 1.27 to 2.26 +/- 1.91, p = 0.047) were increased after exercise intervention, although the change in glucose oxidation rate was not significant. In summary, reduction of non-oxidative glucose disposal may contribute to decreased whole-body glucose utilization. In addition, exercise improves insulin resistance mainly by increasing non-oxidative glucose disposal in type 2 diabetes. PMID- 18410565 TI - Editorial. Symposium on Erythropoiesis and Red Cell Disorders. PMID- 18410566 TI - Genetic variation and susceptibility to infection: the red cell and malaria. AB - There is now convincing evidence that the extremely high frequencies of certain genetic disorders of the red cell involving haemoglobin, the red-cell membrane, or its metabolic pathways reflect relative resistance to malaria over thousands of years. At least some progress has been made towards an understanding of the cellular mechanisms involved, although much remains to be learnt. As well as the extremely valuable information that this field is providing about how exposure to infection has moulded the current structure of the human genome, recent research in this field is starting to provide some valuable new approaches to the better control of parasitic and other infections that remain a major global health problem. PMID- 18410567 TI - Developments in the therapeutic use of erythropoiesis stimulating agents. AB - Recombinant human erythropoietin (rHuEPO) has become the standard therapy for treatment of the anaemia of chronic kidney disease (CKD) during the past two decades. In addition, rHuEPO can be indicated for the treatment of cancer patients on chemotherapy and surgical patients to avoid allogeneic red blood cell transfusion. This review first describes recent attempts in developing rHuEPO congeners (mutated and pegylated rHuEPOs) and mimetics with prolonged half-lives and improved application requirements. Secondly, the pathophysiological background of the regulatory guideline, that blood haemoglobin levels in anaemic CKD or cancer patients on chemotherapy should not be raised above the target value of 120 g/l, is discussed. Finally, potential novel indications are considered for the use of rHuEPO and its analogues as pleiotropic cytoprotectant agents for cardio-, nephro-, hepato- and neuroprotection. PMID- 18410569 TI - Current status of globin gene therapy for the treatment of beta-thalassaemia. AB - beta-Thalassaemia major is a congenital anaemia for which there is presently no curative therapy other than allogeneic haematopoietic stem cell transplantation. This therapeutic option, however, is not available to most subjects for whom there is no available human leucocyte antigen-matched bone marrow donor. The transfer of a regulated globin gene in autologous haematopoietic stem cells is therefore a directly needed alternative treatment. This strategy, simple in principle, raises major challenges in terms of controlling transgene expression, which ideally should be erythroid-specific, differentiation- and stage restricted, elevated, position independent, and sustained over time. Using lentiviral vectors, it has been demonstrated that an optimized combination of proximal and distal transcriptional control elements permits lineage-specific and elevated beta-globin expression in vivo, resulting in the correction of anaemia and secondary organ damage in beta-thalassaemic mice. Several groups have extended these findings to various models of beta-thalassaemia and sickle cell disease. Different globin vectors, however, do not express beta-globin at the same level, and accordingly require the delivery of markedly different vector copy numbers to correct anaemia. Insulators are under investigation to assess whether they might enhance globin gene expression or vector safety. Altogether, recent advances in globin vector design bode well for upcoming clinical trials to assess the therapeutic value of globin gene transfer. PMID- 18410568 TI - The human side of hypoxia-inducible factor. AB - When humans are exposed to hypoxia, systemic and intracellular changes operate together to minimise hypoxic injury and restore adequate oxygenation. Emerging evidence indicates that the hypoxia-inducible factor (HIF) family of transcription factors plays a central regulatory role in these homeostatic changes at both the systemic and cellular levels. HIF was discovered through its action as the transcriptional activator of erythropoietin, and has subsequently been found to control intracellular hypoxic responses throughout the body. HIF is primarily regulated by specific prolyl hydroxylase-domain enzymes (PHDs) that initiate its degradation via the von Hippel-Lindau tumour suppressor protein (VHL). The oxygen and iron dependency of PHD activity accounts for regulation of the pathway by both cellular oxygen and iron status. Recent studies conducted in patients with rare genetic diseases have begun to uncover the wider importance of the PHD-VHL-HIF axis in systems-level human biology. These studies indicate that, in addition to regulating erythropoiesis, the system plays an important role in cardiopulmonary regulation. This article reviews our current understanding of the importance of HIF in human systems-level physiology, and is modelled around the classic physiological response to high-altitude hypoxia. PMID- 18410570 TI - Genetic modifiers of the beta-haemoglobinopathies. AB - Identification of the molecular basis of the beta-thalassaemias and sickle cell disease (SCD) has made it clear that patients with the same beta-globin genotypes can have very variable patterns of clinical expression. Extensive biochemical and pathophysiological studies over the last 50 years have derived two major modifiers--innate ability to produce fetal haemoglobin and co-inheritance of alpha-thalassaemia, subsequently validated by family and population studies. However, these two modifiers do not explain the full clinical spectrum. Genetic studies have been successful in identifying modifiers if the loci have a major clinical effect and if the genetic variants are common. It is possible that additional modifiers could be uncovered using genetic approaches but success will depend on large sample sizes of well-characterised patients with well-defined phenotypes. Since some of the complications, such as overt stroke in SCD, are relatively rare events, intermediate end-points that contribute to the phenotype, such as Transcranial Doppler velocity (a major predictor of stroke in SCD), could be integrated within the genetic analysis. Integrating multiplex genetic testing with clinical and laboratory data to generate predictive models shows potential, but such genetic approaches also require large datasets. PMID- 18410571 TI - Ribosomal dysfunction and inherited marrow failure. AB - Impairment of ribosome biogenesis or function characterizes several of the inherited bone marrow failure syndromes: Diamond-Blackfan anaemia, dyskeratosis congenita (DC), Shwachman-Diamond syndrome and cartilage-hair hypoplasia. These syndromes exhibit overlapping but distinct clinical phenotypes and each disorder involves different aspects of ribosomal biogenesis. The clinical characteristics of each syndrome are briefly reviewed. Molecular studies of ribosome biogenesis and function in each of these syndromes are discussed. Models of how impairment of ribosomal pathways might affect haematopoiesis and tumorigenesis are explored. PMID- 18410572 TI - Studies in haemoglobin E beta-thalassaemia. AB - Haemoglobin E beta-thalassaemia is the commonest form of severe thalassaemia in many Asian countries, but little is known about its natural history, the reasons for its clinical diversity, or its optimal management. Despite its frequency, haemoglobin E beta-thalassaemia is often managed in an ill-defined and haphazard way, usually by demand transfusion. We studied a cohort of Sri Lankan patients with haemoglobin E beta-thalassaemia over 5 years, and identified several genetic and environmental factors possibly contributing to the phenotypic diversity of the disorder. These included modifiers of haemoglobin F production, malaria and age-related changes in adaptation to anaemia. Our findings suggest that in many patients, haemoglobin E beta-thalassaemia can be managed without transfusion, even with low haemoglobin levels. Age-related changes in the pattern of adaptation to anaemia suggest that more cost-effective approaches to management should be explored. PMID- 18410573 TI - Eosinophilic ulcer of the oral mucosa: a distinct entity or a non-specific reactive pattern? AB - Eosinophilic ulcer of the oral mucosa (EUOM) is an uncommon self-limited oral condition that clinically manifests as a solitary ulceration with elevated indurate borders affecting the tongue, buccal mucosa or lip. Microscopically, it is characterized by a polymorphic inflammatory infiltrate with a prominent polymorphonuclear eosinophilic component extending deep into the submucosa, underlying muscle and salivary glands. Large mononuclear cells probably corresponding to histiocytes, myofibroblasts or activated lymphoid cells are also frequently observed. The exact pathogenetic mechanisms implicated in the development of EUOM are poorly understood; however, the possibility that trauma may play a role in its development has been often postulated. Since its original description, the possibility that EUOM could be either considered an individualized disorder or a non-specific reactive pattern secondary to several stimuli has been discussed. EOUM may show some overlapping features with some entities such as atypical histiocytic granuloma, mucosal angiolymphoid hyperplasia with eosinophilia, and Kimura disease. The clinical and histopathological features and the differential diagnosis of EUOM are reviewed and its existence as a distinct disease discussed. PMID- 18410574 TI - Effects of fluorides on Candida albicans. AB - AIMS: To assess whether a short exposure of Candida albicans to commonly used fluorides would affect growth, cell surface hydrophobicity, and adherence to buccal epithelial cells. METHODS: Candida albicans ATCC 90028 and 11 clinical isolates were used. Minimal inhibitory concentrations (MICs) of sodium fluoride (NaF) and of an amine fluoride / stannous fluoride combination (AmF / SnF2) were determined. Yeasts were exposed to MICs of tested agents for 1 h. Subsequently, their growth was recorded spectrophotometrically. Their cell surface hydrophobicity was assessed with n-hexadecane. Adherence to buccal epithelial cells was determined microscopically. Phosphate buffered saline (PBS) and chlorhexidine digluconate (CHX) served as controls. All results were analyzed by one-way ANOVA. RESULTS: MICs of AmF / SnF(2) and CHX varied between 1 and 4 microg ml(-1), whereas those of NaF were 15 000 microg ml(-1). Statistically significant growth inhibition was detected after AmF / SnF(2) (OD(24 h) +/- SD 0.457 +/- 0.059) and CHX (0.175 +/- 0.065) in comparison with PBS (0.925 +/- 0.087) and NaF (0.813 +/- 0.081). All strains demonstrated uniform behavior. Only minor changes in cell surface hydrophobicity and adherence to buccal epithelial cells (BEC) were detected. CONCLUSION: Growth inhibition of AmF / SnF(2) was comparable with that of CHX whereas NaF had a weaker effect. Exposure to the fluorides did not seem to alter the cell surface hydrophobicity nor adherence to BEC. PMID- 18410575 TI - Analysis of appositional bone formation using a novel rat experimental model. AB - AIM: To analyze the process of appositional bone formation using our original rat experimental model. MATERIALS AND METHODS: Rats were anesthetized and a ring made of polytetrafluorethylene was placed on the parietal bone surface in the surgical procedure. The time course of appositional bone formation was analyzed with histomorphometry and in situ hybridization for type I collagen and bone sialoprotein. RESULTS: The rat experimental model allowed new bone to be formed on the pre-existing bone surface and persist for 12 weeks. We demonstrated that bone is apposed actively for the first 4 weeks and less actively thereafter. CONCLUSIONS: The experimental model may contribute to biological analysis for appositional bone formation expected to occur in clinical procedures such as alveolar bone augmentation and sinus lifting. PMID- 18410576 TI - Secreted protein acidic, rich in cysteine induces pulp cell migration via alphavbeta3 integrin and extracellular signal-regulated kinase. AB - AIM: The aim of this study was to investigate the influence of secreted protein acidic, rich in cysteine (SPARC) on the migration of human dental pulp (HDP) cells. METHODS: Secreted protein acidic, rich in cysteine was applied in the lower chamber of the chemotaxis apparatus and migration was determined by counting the cells that migrated through the membrane. To determine the signaling pathway involved, cells were incubated with inhibitors for 30 min prior to the migration assay. RESULTS: The results indicated that SPARC induced HDP cell migration in a dose-dependent manner via extracellular signal-regulated kinase (ERK). The migration could be inhibited both by the anti-alphavbeta3 integrin antibody and by suramin, a non-selective growth factor receptor and G-protein coupled receptor antagonists. The anti-alphavbeta3 integrin antibody could also inhibit ERK activation, suggesting the possible role of alphavbeta3 integrin on the regulation of ERK and cell migration. Interestingly, both suramin and SB225002, another G-protein coupled receptor antagonist, suppressed ERK activation. CONCLUSIONS: Secreted protein acidic, rich in cysteine could act as a chemotactic factor and facilitate migration, possibly through the G-protein coupled receptor, alphavbeta3 integrin and ERK. The data support that SPARC could play a crucial role in dental pulp tissue repair by inducing dental pulp cell migration. PMID- 18410577 TI - Patterns of oral manifestation of HIV/AIDS among 225 Nigerian patients. AB - OBJECTIVE: The purpose of this study was to determine the prevalence of the oral manifestations of HIV/AIDS and to correlate the prevalence of these lesions with the stages of the disease in the Ife-Ijesa zone, Nigeria. No comprehensive data were available for correlating it with the staging of HIV/AIDS in this region. SUBJECTS: The pattern of oral HIV lesions as classified by the EC-Clearinghouse was studied in 225 confirmed consecutive HIV-infected patients in this zone. METHODS: Clinical dental examinations were conducted under natural daylight on all consenting HIV patients, sitting in an upright chair, using dental mirrors and probes. RESULTS: The prevalence of oral HIV lesions was 84.0%, with lesions ranging in number from one to six. The commonest HIV lesion was pseudo-membranous candidiasis (43.1%) followed by erythematous candidiasis (28.9%), angular cheilitis (28.9%), linear gingival erythema (24.0%) and ulcerations (8.9%). Lesions less commonly found were oral hairy leukoplakia (1.3%) and salivary gland swellings (1.3%). Heterosexual intercourse was the most common mode of transmission (94.7%) and HIV-1 (96.9%) the most prevalent pathogen among the study population. The majority of the patients were in the WHO clinical stage III (59.1%) and presented late. CONCLUSIONS: The prevalence of oral HIV lesions in the present study was high. PMID- 18410578 TI - The prevalence of oral mucosal lesions in adults from the Turin area. AB - AIM: To assess the prevalence of oral mucosal lesions (OML) and evaluate its association with tobacco and alcohol consumption and the wearing of removable dentures in an adult population from the Turin area, Italy. MATERIALS AND METHODS: A retrospective study, based on an invitational self-selected screening, was performed on 4098 subjects. It included clinical examination plus biopsies when necessary. Patient history included age, sex, denture wearing and risk habits. Internationally accepted criteria were adopted to classify the OMLs. RESULTS: Males were observed to have more OMLs (557/2040; 27.3%vs 471/2058; 22.89%). Overall OML prevalence was linked to risk habits and age. Tobacco was linked to leukoplakia, melanin pigmentation, smoker's palate, frictional lesions and papilloma. It was negatively related to recurrent aphthous stomatitis and oral lichen planus. Alcohol was linked to leukoplakia, frictional lesions and median rhomboid glossitis. The tobacco-alcohol association was linked to frictional lesions, leukoplakia, melanin pigmentation and smoker's palate. Denture wearers had an overall higher prevalence of OMLs, in particular candidiasis, traumatic and frictional lesions. CONCLUSIONS: The prevalence of OMLs in Turin seems to be comparable to those in other European studies and emphasize that risk habits and denture wearing have some relationship with the presence of OMLs. PMID- 18410579 TI - Scope of practice, referral patterns and lesion occurrence of an oral medicine service in Australia. AB - AIM: The purpose of this study was to examine the scope of practice, lesion occurrence and utilisation of referral-based hospital and private practice oral medicine and oral pathology (OMP) services in Australia. MATERIALS AND METHODS: Clinical records of patients referred to a hospital (n=500) and private (nbequals;1104) OMP clinic were audited. For each patient, the following parameters were recorded: age, gender, source of referral, reason for referral, site of lesion/condition if applicable, medical and drug history, diagnostic services utilised, clinical and histopathological diagnoses rendered, medications prescribed and further treatment required. RESULTS: A majority of the referrals were generated by general dental practitioners. The most commonly seen problems were epithelial hyperplasia/hyperkeratosis, oral candidosis, oral lichen planus, xerostomia, recurrent aphthous ulcers and burning mouth syndrome. OMP specialists requested diagnostic imaging for 13% of hospital and 9.42% of private patients, diagnostic biopsies were required for 18.4% of hospital and 19.3% of private patients, blood tests were ordered for 14.4% of hospital and 12.13% of private patients, while medications were prescribed for approximately 36% of hospital and 51% of private patients. CONCLUSIONS: This study is the first to detail the scope of practice, lesion occurrence and utilisation of services offered by OMP specialists in Australia. The demand for OMP services is strong. PMID- 18410580 TI - Fatty acid synthase expression in squamous cell carcinoma of the tongue: clinicopathological findings. AB - BACKGROUND: Overexpression of fatty acid synthase (FAS), the cytosolic enzyme responsible for the conversion of dietary carbohydrates to fatty acids, has been reported in several human malignancies and pointed as a potential prognostic marker for some tumors. This study investigated whether FAS immunohistochemical expression is correlated with the clinicopathological characteristics of oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: The clinical features of 102 patients with OSCC of the tongue treated in a single institution were obtained from the medical records and all histopathological diagnoses were reviewed. The expression of FAS was determined by the standard immunoperoxidase technique in formalin-fixed and paraffin-embedded specimens and correlated with the clinicopathological characteristics of the tumors. RESULTS: Eighty-one cases (79.41%) were positive for FAS. Microscopic characteristics such as histological grade (P < 0.05), lymphatic permeation (P < 0.001), perineural infiltration (P < 0.05), and nodal metastasis (P < 0.02) were associated with FAS status. A significantly lower survival probability for patients with advanced clinical stage (log-rank test, P < 0.001), lymph nodes metastasis (log-rank test, P < 0.001), presence of vascular permeation (log-rank test, P = 0.05), and perineural invasion (log-rank test, P = 0.01) was observed in the studied samples. CONCLUSION: The expression of FAS in OSCC of the tongue is associated with the microscopic characteristics that determine disease progression and prognosis. PMID- 18410581 TI - Exposure confounding causes dominance of antiplasmodial IgG3 subclass antibody responses in uncomplicated malaria. PMID- 18410582 TI - Statistical consideration of the role of potential confounders on the association between immunological responses and disease. PMID- 18410583 TI - Melatonin, consciousness, and traumatic stress. AB - Descartes intuitively anticipated the so-called 'binding problem' of consciousness and thought that the pineal gland enables spatio-temporal integration in cognitive processing. Recent findings indicate that a major role in the process of temporal integration and binding involve neurons in suprachiasmatic nuclei, specifically targeting the pineal gland and other structures, and control the neuroendocrine rhythms. Melatonin is an endocrine output signal of the clock and provides circadian information as an endogenous synchronizer which stabilizes and reinforces circadian rhythms. This integrative process occurs at the different levels of the circadian network via gene expression in some brain regions and peripheral structures that enables integration of circadian, hormonal, and metabolic information and creating temporal order of bodily and mental experience. This specific temporal order is reflected in associative sequentiality that is necessary for cognition, behavior and all processes of memory consolidation that must preserve all information in the temporal causal order and synchrony. In this context, recent findings suggest that melatonin could be a potential regulator in the processes that contribute to memory formation, long-term potentiation, and synaptic plasticity in the hippocampus and other brain regions. There is evidence that stress disrupts normal activity and memory consolidation in the hippocampus and prefrontal cortex, and this process leads to memories that are stored without a contextual or spatiotemporal frame. These findings emphasize a specific role of melatonin in mechanisms of consciousness, memory and stress and are also consistent with reported studies that indicate melatonin alterations under stressful conditions and in mental disorders. PMID- 18410584 TI - Effect of oral melatonin on the procoagulant response to acute psychosocial stress in healthy men: a randomized placebo-controlled study. AB - Acute mental stress is a potent trigger of acute coronary syndromes. Catecholamine-induced hypercoagulability with acute stress contributes to thrombus growth after coronary plaque rupture. Melatonin may diminish catecholamine activity. We hypothesized that melatonin mitigates the acute procoagulant stress response and that this effect is accompanied by a decrease in the stress-induced catecholamine surge. Forty-five healthy young men received a single oral dose of either 3 mg melatonin (n = 24) or placebo medication (n = 21). One hour thereafter, they underwent a standardized short-term psychosocial stressor. Plasma levels of clotting factor VII activity (FVII:C), FVIII:C, fibrinogen, D-dimer, and catecholamines were measured at rest, immediately after stress, and 20 min and 60 min post-stress. The integrated change in D-dimer levels from rest to 60 min post-stress differed between medication groups controlling for demographic and metabolic factors (P = 0.047, eta(p)(2) = 0.195). Compared with the melatonin group, the placebo group showed a greater increase in absolute D-dimer levels from rest to immediately post-stress (P = 0.13; eta(p)(2) = 0.060) and significant recovery of D-dimer levels from immediately post-stress to 60 min thereafter (P = 0.007; eta(p)(2) = 0.174). Stress-induced changes in FVII:C, FVIII:C, fibrinogen, and catecholamines did not significantly differ between groups. Oral melatonin attenuated the stress-induced elevation in the sensitive coagulation activation marker D-dimer without affecting catecholamine activity. The finding provides preliminary support for a protective effect of melatonin in reducing the atherothrombotic risk with acute mental stress. PMID- 18410585 TI - Regression of herpes viral infection symptoms using melatonin and SB-73: comparison with Acyclovir. AB - Infection with Herpes simplex virus type 1 (HSV-1) typically causes lesions of the mouth, face, skin, esophagus, or brain. Herpes simplex virus type 2 (HSV-2) usually causes infections of the genitals, rectum, skin, hands, or meninges. The herpes viruses are a major cause of blindness from keratitis. The usual drugs used for herpes are Vidarabine, Acyclovir, Penciclovir and Ganciclovir; they are associated with several complications. The aim of this study was to investigate if a formulation containing 2.5 mg melatonin and 100 mg SB-73 would help patients with herpes, and to compare the preparation with 200 mg Acyclovir. SB-73 is a mixture of magnesium, phosphate, fatty acids extracted from Aspergillus sp. which has anti-herpes virus properties. A single blind randomized study was performed in which 70 patients underwent treatment using the supplement cited above (group A) and 75 received treatment of 200 mg Acyclovir (group B). Sixty-seven patients of the group A (95.7%) reported a complete regression of symptoms after 7 days of treatment. By comparison, 64 subjects (85.3%) of the Acyclovir reported regression of symptoms in the same period. There was statiscally significant difference between the groups (P < 0.05). PMID- 18410587 TI - Combinational polymorphisms of four DNA repair genes XRCC1, XRCC2, XRCC3, and XRCC4 and their association with oral cancer in Taiwan. AB - BACKGROUND: Many single nucleotide polymorphisms (SNPs) have been found to be associated with oral cancer but the biological interactions through SNPs are seldom addressed. In this study, we focused on the joint effect for SNP combinations of four DNA repair genes, X-ray repair cross-complementing groups (XRCCs) 1-4, involved in major cancer-related pathways. METHODS: Single nucleotide polymorphism genotyping was determined using by polymerase chain reaction-restriction fragment length polymorphism in this study (case = 103, control = 98). Different numbers of combinational SNPs with genotypes called the pseudo-haplotypes from these chromosome-wide genes were used to evaluate their joint effect on oral cancer risk. RESULTS: Except for XRCC2 rs2040639-AG, none of these SNPs was found to individually contribute to oral cancer risk. However, for two combined SNPs, the proportion of subjects with oral cancer was significantly higher in the pseudo-haplotype with AG-CC genotypes in rs2040639-rs861539 (XRCC2 XRCC3) compared with those with non-AG-CC genotypes. Similarly, the pseudo haplotype of rs2040639-rs861539-rs2075685 (XRCC2-XRCC3-XRCC4) and rs2040639 rs861539-rs2075685-rs1799782 (XRCCs 1-4) with specific genotype pattern (AG-CC-TG and CT-AG-CC-TG) among three and four combinational SNPs were significantly associated with oral cancer. After controlling for age, gender, smoking, drinking, and betel nut chewing, the estimated odds ratio of oral cancer were 2.45, 5.03, and 10.10 for two, three and four specific SNP combinations, respectively, comparing these specific pseudo-haplotypes to their corresponding non-pseudo-haplotypes. CONCLUSION: We have identified the potential combined XRCCs 1-4 SNPs with genotypes that were associated with oral cancer risk and may have an impact on identification of a high-risk population. PMID- 18410589 TI - Introduction: atherosclerosis as inflammation: a controversial concept becomes accepted. PMID- 18410586 TI - Melatonin represses oxidative stress-induced activation of the MAP kinase and mTOR signaling pathways in H4IIE hepatoma cells through inhibition of Ras. AB - Reactive oxygen species (ROS) have been implicated in the pathogenesis of a variety of diseases, and antioxidant treatment is currently being investigated as a potential therapy to attenuate the detrimental effects of ROS-mediated oxidative stress. Melatonin is a potent naturally produced antioxidant, which acts through various mechanisms to ameliorate the toxic effects of ROS. However, little is known about the mechanisms of signaling pathways through which melatonin acts to reverse the effects of ROS. In the present study, the effect of melatonin treatment on the hydrogen peroxide (H(2)O(2))-induced activation of the mitogen-activated protein kinase (MAPK) and mammalian target of rapamycin (mTOR) signaling pathways was assessed in H4IIE hepatoma cells. It was found that melatonin strongly attenuated H(2)O(2)-induced activation of the ERK1/2 and p38 MAP kinases, as well as several of their downstream targets. Melatonin also attenuated the H(2)O(2)-induced phosphorylation of Akt and the Akt substrate mTOR, as well as a downstream target of mTOR action, 4E-BP1. Upregulation of ERK1/2, p38, and Akt signaling by H(2)O(2) was accompanied by activation of Ras, an effect that was blocked by melatonin. Overall, the results suggest that melatonin acts to prevent many of the H(2)O(2)-induced alterations in the MAPK and mTOR signaling pathways through inhibition of Ras, at least in H4IIE hepatoma cells. PMID- 18410590 TI - Autoimmunity in atherosclerosis: a protective response losing control? AB - Atherosclerosis is a chronic inflammatory disease characterized by accumulation of oxidized lipoproteins, increased cell death and hypertrophic degeneration of the arterial intima. The disease process is associated with local formation of modified self antigens that are targeted by both innate and adaptive immune responses. Although it remains to be firmly established it is likely that these autoimmune responses initially have a beneficial effect facilitating the removal of potentially harmful rest products from oxidized LDL and dying cells. However, studies performed on hypercholesterolaemic mice deficient in different components of the immune system uniformly suggest that the net effect of immune activation is pro-atherogenic and that atherosclerosis, at least to some extent, should be regarded as an autoimmune disease. These observations point to the possibility of developing new treatments for atherosclerosis based on modulation of immune responses against plaque antigens, an approach presently tested clinically for several other chronic inflammatory diseases with autoimmune components. Pilot studies in animals have provided promising results for both parental and oral vaccines based on oxidized LDL antigens. The time when this concept is ready for clinical testing is rapidly approaching but it will be important not to underestimate the difficulties that will be encountered in transferring the promising results from experimental animals into humans. PMID- 18410591 TI - Oxidation-specific epitopes are important targets of innate immunity. AB - During the oxidation of LDL, a central pathophysiological component of atherogenesis, a wide variety of chemical and physical changes occur leading to the generation of oxidation-specific neoepitopes. These epitopes are not only immunogenic, leading to adaptive humoral responses, but are also a prominent target of multiple arcs of innate immunity. The pattern recognition receptors (PRRs) of innate immunity are germ line encoded, conserved by natural selection, and bind to pathogen-associated molecular patterns (PAMPs) common on multiple structures. However, it is not intuitive as to why they should recognize oxidation-specific neoepitopes. Yet it is clear that multiple macrophage scavenger receptors, which are classic PRRs, recognize oxidation-specific epitopes, such as those found on oxidized LDL (OxLDL). Other innate proteins, such as C-reactive protein, also bind to OxLDL. Natural antibodies (NAbs), the humoral arc of innate immunity, provide a nonredundant role in the first line of defence against pathogens, but are also believed to provide important homeostatic house-keeping functions against self-antigens. Our work demonstrates that oxidation-specific epitopes, as found on OxLDL, are a major target of NAbs. In this review, we will discuss the specific example of the prototypic NAb T15/E06, which is increased in atherosclerotic mice and mediates atheroprotection, and discuss the potential role of NAbs in atherogenesis, and in inflammation in general. We also review data that oxidation-specific epitopes are generated whenever cells undergo programmed cell death, forming a common set of PAMPs recognized by oxidation-specific PRRs on macrophages, NAbs and innate proteins. We present the hypothesis that oxidation-specific epitopes on apoptotic cells exerted evolutionary pressure for the conservation of these PRRs and also serve to maintain the expansion of a substantial proportion of NAbs directed to these stress-induced self-antigens. PMID- 18410592 TI - Regulatory T-cell immunity and its relevance to atherosclerosis. AB - Atherosclerosis is a chronic inflammatory disease of the arterial wall where both innate and adaptive immune responses contribute to disease initiation and progression. Recent studies from several groups suggest that subtypes of T cells, called regulatory T cells, previously shown to maintain immunological tolerance, inhibit the development and progression of atherosclerosis. Here, we review the current knowledge on the regulatory T-cell response and the major cytokines involved in its modulation in the context of atherosclerosis. PMID- 18410593 TI - Microsomal prostaglandin E synthase-1 inhibition in cardiovascular inflammatory disease. AB - Prostaglandins (PGs), particularly PGE2 and prostacyclin (PGI2), are potent mediators of pain and inflammation. Both atherosclerosis and aortic aneurysm exhibit the hallmarks of inflammation. However, randomized trials of inhibitors of PG synthesis--nonsteroidal anti-inflammatory drugs--reveal that they predispose to cardiovascular risk. This appears to be consequent to inhibition of PGI2 and PGE2 formed by cyclooxygenase-2 (COX-2). Inhibitors of microsomal PGE synthase-1 (mPGES-1) are being developed for relief of pain and interest has focused on their potential impact on the cardiovascular system. Deletion of mPGES 1 retards atherogenesis and limits aortic aneurysm formation in hyperlipidaemic mice. However, it does not predispose to thrombogenesis and has a limited impact on blood pressure compared to inhibition of COX-2. This occurs despite the potential of the suppressed PGE2 in affording cardioprotection via its EP2 and EP4 receptors. However, deletion of mPGES-1 permits rediversion of the PGH2 substrate to other PG synthases and augmented formation of PGI2 and PGD2 mitigates this effect. However, increased PGI2 may also attenuate relief of pain. Pain relief seems likely to be a nuanced indication for mPGES-1 inhibitors, but they have therapeutic potential in syndromes of cardiovascular inflammation, cancer and perhaps in neurodegenerative disease. However, as the products of substrate rediversion vary according to cell type, these drugs may have contrasting impact amongst individuals at varied stages of disease evolution. PMID- 18410594 TI - From vulnerable plaque to atherothrombosis. AB - Plaque rupture precipitates approximately 75% of all fatal coronary thrombi. Therefore, the plaque prone to rupture is the primary focus of this review. The lipid-rich core and fibrous cap are pivotal in the understanding of plaque rupture. Plaque rupture is a localized process within the plaque caused by degradation of a tiny fibrous cap rather than by diffuse inflammation of the plaque. Atherosclerosis is a multifocal disease, but plaques prone to rupture seem to be oligofocal at most. PMID- 18410596 TI - Role of microparticles in atherothrombosis. AB - Cell activation or apoptosis leads to plasma membrane blebbing and microparticle (MP) release in the extracellular space. MPs are submicron membrane vesicles which express a panel of phospholipids and proteins specific of the cells they are derived from. Exposure of negatively charged phospholipids and tissue factor confers a procoagulant potential to MPs. MPs accumulate in the lipid core of the atherosclertotic plaque and is a major determinant of its thrombogenecity. Elevation of plasma MPs levels, particularly those of endothelial origin, reflects cellular injury and is considered now as a surrogate marker of vascular dysfunction. Thus, MPs can be seen as triggers of a vicious circle for they promote prothrombogenic and pro-inflammatory responses as well as cellular dysfunction within the vascular compartment. A better knowledge of MP composition and biological effects as well as the mechanisms leading to their clearance will probably open new therapeutic approaches in the treatment of atherothrombosis. PMID- 18410597 TI - Identifying the susceptibility genes for coronary artery disease: from hyperbole through doubt to cautious optimism. AB - The genetic basis of coronary artery disease (CAD) is complex, and the fact that an alarmingly high proportion of reported associations between genetic variants and CAD are not replicated has generated uncertainty as to whether molecular genetics is ever going to deliver on the promises delivered in the late 1990s. However, during 2007, the first generation of large-scale genome-wide association studies using high-density, single nucleotide polymorphism genotyping arrays have revealed genetic variants that are robustly associated with CAD and CAD-related traits such as type 2 diabetes and obesity. In particular, a robust susceptibility locus for CAD has been identified on chromosome 9p21. Also, evidence has been obtained that multiple rare alleles with fairly strong phenotypic effects may contribute to the genetic heritability of CAD, in addition to common variants with a modest impact on risk. Furthermore, new mechanistic connections have been discovered between different common complex diseases including CAD. This review focuses on the challenges and recent advances of molecular genetics in dissecting the molecular pathophysiology of atherothrombosis and defining novel targets for treatment. PMID- 18410595 TI - The molecular mechanisms of the thrombotic complications of atherosclerosis. AB - Our evolving knowledge of the cellular and molecular mechanisms underlying atherosclerosis has helped uncover the underlying causes behind thrombotic complications of this disease. Most fatal coronary thrombosis result from fibrous cap rupture or superficial erosion. Recent research has established a role for matrix metalloproteinases in the regulation of aspects of plaque structure related to propensity to disrupt and provoke thrombosis. Inflammatory pathways impinge on proteinase activity and aspects of oxidative stress that may favour plaque disruption. Novel molecular imaging strategies may permit visualization of proteinase activity in vivo, providing a new functional window on pathophysiology. PMID- 18410598 TI - Proteolysis of low density lipoprotein particles by Porphyromonas gingivalis microorganisms: a novel biochemical link between periodontitis and cardiovascular diseases? PMID- 18410599 TI - Hepatitis B and fatty liver: causal or coincidental? PMID- 18410600 TI - Antiviral therapy for various stages of HBV-related diseases: lamivudine and beyond. PMID- 18410601 TI - Chemotherapy for advanced hepatocellular carcinoma. PMID- 18410602 TI - Role of magnetic resonance cholangiopancreatography and other non-invasive imaging modalities in assessing bile duct stones. PMID- 18410603 TI - Liver stem cells: a scientific and clinical perspective. AB - The promise of liver stem cells lie in their potential to provide a continual and readily available source of liver cells that can be used for gene therapy, cellular transplant, bioartificial liver-assisted devices, drug toxicology testing and use as an in vitro model to understand the developmental biology of the liver. Both the rodent and human embryonic stem cell, bone marrow hematopoietic stem cell, mesenchymal stem cell, umbilical cord blood cell, fetal liver progenitor cell, adult liver progenitor cell as well as the mature hepatocyte have been reported to be capable of self-renewal, giving rise to daughter hepatocytes both in vivo and in vitro. These cells can repopulate livers in animal models of liver injury and seemingly improve liver function. However, significant challenges still exist before these cells can be used in humans. These include lack of consensus in immunophenotype of liver progenitor cells, uncertainty of the physiological role of reported candidate stem/progenitor cell, practicality in obtaining sufficient quantity of cells for clinical use and concerns over ethics, long-term efficacy and safety. Current molecular techniques of stem cell identification are confounded by cell fusion, horizontal gene transfer, incomplete differentiation and fetal microchimerism. Reports of stem cell transplantation and phase 1 trials of bone marrow transplantation in humans for liver diseases are exciting but require more robust verification. We review the evidence for various candidate stem cells, human clinical trials reported to date and highlight the challenges facing clinicians in their quest to use liver stem cells to save lives. PMID- 18410604 TI - Current treatment of HIV/hepatitis B virus coinfection. AB - Coinfection with HIV and hepatitis B virus (HBV) has become a significant global health problem. Liver disease is now one of the leading causes of morbidity and mortality in individuals with HIV, particularly those with viral hepatitis. There are a number of agents available with dual activity against HIV and HBV, and effective treatment depends on understanding the potential advantages and pitfalls in using these agents. There are a number of unresolved issues in the management of HIV/HBV coinfection. These include the role of liver biopsy, the significance of normal aminotransferase levels, serum HBV DNA threshold for treatment, treatment end-points, and the treatment of HBV when HIV does not yet require treatment. Treatment of HBV should be considered in individuals with HIV/HBV coinfection with evidence of significant fibrosis (>/=F2), or with elevated serum HBV DNA levels (>2000 IU/mL). Sustained suppression of serum HBV DNA to below the level of detection by the most sensitive available assay should be the goal of therapy, and, at present, treatment of HBV in HIV/HBV coinfection is lifelong. If antiretroviral therapy is required, then two agents with anti-HBV activity should be incorporated into the regimen. If antiretroviral therapy is not required, then the options are pegylated interferon, adefovir or the early introduction of antiretroviral therapy. Close monitoring is necessary to detect treatment failure or hepatic flares, such as immune reconstitution disease. Further studies of newer anti-HBV agents in individuals HIV/HBV coinfection may advance treatment of this important condition. PMID- 18410605 TI - Columnar-lined esophagus: time to drop the eponym of "Barrett": Historical review. AB - There can be few medical conditions that have been surrounded by as much confusion about their definition or terminology as columnar-lined esophagus (CLE); approximately 30 different terms and eponyms have been used to describe this condition. The history of this condition can be divided into five stages: (i) descriptive stage, 1906-1950; (ii) "argument" stage, 1950-1963; (iii) "significant" stage, 1963-1973; (iv) surveillance stage, 1973-1990; and (v) refined research stage, 1990-present. The use of the eponym "Barrett's" to describe CLE is not justified from a historical point of view. Lining of the lower esophagus by columnar epithelium was termed "Barrett's esophagus" after the presentation by Barrett in 1957. Although this finding has been attributed to Barrett, the work of others, including Tileston, Lortat-Jacob, and Allison and Johnstone, preceded Barrett's description. The historical aspects of CLE were reviewed to show how little Norman Barrett had contributed to the core concept of this condition in comparison to the contributions of other investigators, particularly the contribution of Philip Allison. Based on many discussed historical facts, we are not in favor of retaining the term "Barrett's esophagus" and we propose that CLE be henceforth referred to as "columnar-lined esophagus". PMID- 18410606 TI - Gastroesophageal reflux disease in Asia: a condition in evolution. AB - Widespread epidemiological changes, rising prevalence and gradual shifts in patterns of disease manifestations: this is the changing face of gastroesophageal reflux disease (GERD) in Asia. Are we witnessing a disease in evolution or merely the result of increased and more accurate case reporting that comes with advancing diagnostic technology, better medical facilities and heightened awareness of the disease? Do the figures reported really reflect the actual scenario or is there more to it than meets the eye? In this article, we take you back in time to review relevant developments over the past decade or so. We will draw on findings from across Asia, take an in-depth look at prevailing trends and patterns and examine some of the most plausible explanations behind the dynamics of this epidemiological transition. PMID- 18410607 TI - Randomised comparison of the FerX Ella antireflux stent and the ultraflex stent: proton pump inhibitor combination for prevention of post-stent reflux in patients with esophageal carcinoma involving the esophago-gastric junction. AB - BACKGROUND AND AIM: Metal stents placed across the gastroesophageal junction in patients with malignant dysphagia frequently present with reflux symptoms. We compared an antireflux stent with a standard open stent used in combination with proton pump inhibitor medication. METHODS: Forty-nine patients with dysphagia due to inoperable carcinoma in the lower third of the esophagus were randomly selected to receive either a antireflux valve stent (FerX-Ella) (n = 22) or a covered standard open stent (Ultraflex), which was combined with proton pump inhibitors such as omeprazole (n = 26). The technical success, the presence of reflux, and complications were recorded. RESULTS: Reflux was seen in 3/22 patients (13.6%) in the FerX-Ella group and in 2/26 patients (7.7%) in the Ultraflex and proton pump inhibitor combination group (P-value not significant). In both groups, a significant improvement in the dysphagia score was seen and no statistically significant difference was detected between the two groups (P = 0.84). The FerX-Ella stents migrated more frequently (32%) than the Ultraflex stents (23%). This also necessitated surgical intervention more frequently in the FerX-Ella group (2/22, 9.1%) compared to the Ultraflex group (1/26, 3.8%). CONCLUSION: The antireflux stent had no demonstrable advantages compared to the combination of standard open stent and proton pump inhibitor medication. PMID- 18410608 TI - Combination therapy of thymosin alpha-1 and lamivudine for HBeAg positive chronic hepatitis B: A prospective randomized, comparative pilot study. AB - BACKGROUND AND AIM: Monotherapy of lamivudine, interferon-alpha (IFN-alpha), and thymosin alpha-1 (Talpha1) is unlikely to be sufficient for the eradication of a chronic hepatitis B virus (HBV) infection. The aim of our study is to elucidate whether the combination of Talpha1 and lamivudine is superior to lamivudine monotherapy in hepatitis B e antigen (HBeAg) positive naive patients with chronic hepatitis B. METHODS: Sixty-seven patients were assigned to two different groups in a randomized manner. The combination group (n = 34) received Talpha1 (1.6 mg subcutaneously, twice a week) and lamivudine (100 mg orally, daily) for 24 weeks, followed by continuous lamivudine therapy. The monotherapy group (n = 33) received lamivudine monotherapy continuously. RESULTS: The incidence of HBeAg seroconversion at 24 weeks was 26.5% (9/34) in the combination group and 6.1% (2/33) in the monotherapy group (P = 0.024). However, there was no statistically significant difference between 26.5% (9/34) in the combination group and 12.1% (4/33) in the monotherapy group at 52 weeks (P = 0.138). The emergence of viral breakthrough gradually increased to 35.3% (12/34) in the combination group, and to 21.2% (7/33) in the monotherapy group at 52 weeks (P = 0.201). CONCLUSIONS: The combination treatment of Talpha1 and lamivudine did not have an obvious benefit of virological and biochemical response as compared to the lamivudine monotherapy during the combination period. In addition, after the cessation of Talpha1 treatment, the combination therapy did not prevent the occurrence of viral and biochemical breakthroughs. PMID- 18410609 TI - Early activation of mucosal dendritic cells and macrophages in acute Campylobacter colitis and cholera: An in vivo study. AB - BACKGROUND AND AIM: Macrophages and dendritic cells are closely related mononuclear phagocytic cells. Little is known about their in vivo role in acute intestinal bacterial infections in humans. We undertook to evaluate these cells in rectal mucosal biopsies of patients with acute colitis. METHODS: All mucosal mononuclear phagocytic cells in rectal biopsies of patients with acute Campylobacter colitis (n = 5), shigellosis (n = 5), and cholera (n = 10) were evaluated ultrastructurally and compared with those in controls (n = 5). RESULTS: Mononuclear phagocytic cells in the superficial rectal mucosa showed a higher prevalence of ultrastructural features of activation in Campylobacter colitis and cholera than in controls. A lower prevalence of features of activation with increased monocytes was seen in shigellosis. Cells with the ultrastructural morphology of activated dendritic cells constituted 41% and 45% of all mononuclear phagocytic cells in two of five patients with Campylobacter colitis and 4-22% of cells in four of 10 patients with cholera. Their presence in patients with Campylobacter colitis was associated with significant surface epithelial damage and prominent acute inflammatory changes in the mucosa. CONCLUSIONS: This is the first ultrastructural study to show activated macrophages and dendritic cells in vivo in acute Campylobacter colitis and cholera. Dendritic cell activation occurred early in the clinical course of these infections. Surface epithelial damage may play a role in the activation of dendritic cells. PMID- 18410610 TI - Enhanced expression of mucin 6 glycoprotein in cholangiocarcinoma tissue from patients in Thailand as a prognostic marker for survival. AB - BACKGROUND AND AIM: Cholangiocarcinoma (CCA) is a mucin-producing cancer that has poor prognosis. Mucin 6 (MUC6) is a mucin that is normally co-expressed with the trefoil factor family-2 (TFF2) trefoil peptide. Both MUC6 and TFF2 have been reported to be involved in the progression of many types of cancers. The aim of this study was to determine the expression of MUC6 and TFF2 in CCA tissues and associate these results with clinical data. METHODS: MUC6 and TFF2 were detected in CCA tissues by immunohistochemistry. The correlations of MUC6 and TFF2 expressions with clinical data were analyzed. RESULTS: We determined the significant co-expression of both proteins in serial CCA tissues. The high expressions of MUC6 and TFF2 were demonstrated in 37% and 31% of patients, respectively. The expression levels decreased in the advanced stage of CCA when clinical metastasis was exhibited. The high expression of either protein showed a correlation with prolonged postoperative survival time, but only a high expression of MUC6 is significantly correlated with a 5-year survival rate. A multivariate Cox regression analysis revealed that a low expression of MUC6, high expression of TFF2, age of patients >56 years, tumor size >5 cm, and poorly differentiated histological type were independent, poor prognostic indicators for CCA. CONCLUSION: MUC6 showed a good correlation with the survival of CCA patients. It may be of value to propose that MUC6 is a good prognostic marker for CCA management. PMID- 18410611 TI - Detection of HBV core promoter and precore mutations helps distinguish flares of chronic hepatitis from acute hepatitis B. AB - BACKGROUND AND AIM: Acute exacerbation of chronic hepatitis B has to be distinguished from acute hepatitis, because treatment strategies differ between them. METHODS: Mutations in the core promoter and precore region of hepatitis B virus (HBV) were determined in 36 patients with acute exacerbation of chronic hepatitis B, in whom alanine aminotransferase (ALT) increased above 500 IU/L, as well as the 36 patients with acute hepatitis. RESULTS: Mutations in the core promoter (A1762T/G1764A) and precore region (G1896A) were more frequent in patients with acute exacerbation of chronic hepatitis than acute hepatitis (81% vs 19%; P < 0.0001 and 58% vs 6%; P < 0.0001, respectively). Of the 19 patients with mutations in both the core promoter and precore region, 17 (89%) had acute exacerbation of chronic hepatitis. In contrast, among the 32 patients with the wild-type for both the core promoter and precore region, 29 (89%) developed acute hepatitis. By multivariate analysis, the double mutation in the core promoter was predictive of acute exacerbation in chronic hepatitis with the highest odds ratio at 26.4. CONCLUSIONS: In patients with hepatitis B having ALT levels >500 IU/L, mutations in the core promoter and precore region are useful in distinguishing acute exacerbation of chronic from acute HBV infection. Detection of these mutations would be useful for commencing prompt antiviral treatments on patients with acute exacerbation of chronic hepatitis for a better prognosis. PMID- 18410612 TI - Education of imaging. Hepatobiliary and pancreatic: biliary stent causing imminent duodenal perforation. PMID- 18410613 TI - Education of imaging. Gastrointestinal: post-operative esophageal fistula. PMID- 18410615 TI - Education of imaging. Hepatobiliary and pancreatic: adenosquamous carcinoma of the pancreas. PMID- 18410614 TI - Education of imaging. Gastrointestinal: diagnostic dilemma of tattoed small bowel. PMID- 18410616 TI - Education of imaging. Hepatobiliary and pancreatic: iatrogenic hemobilia. PMID- 18410617 TI - Malignant nodular hidradenoma of the skin: report of seven cases. AB - BACKGROUND: Malignant nodular hidradenoma (MNH) is an infrequent, highly malignant, primary skin tumour derived from eccrine sweat glands. Most tumours occur in elderly individuals. MNH has very poor prognosis, high recurrence and a high rate of metastases. The best method of treatment is still unclear: radical surgical excision is widely used, and selective lymph node dissection is also suggested. The value of the adjuvant radiotherapy and chemotherapy has not been confirmed. PATIENTS AND TREATMENT: Seven MNH patients (4 men, 3 women, age 60-87 years) were treated between 1991 and 2007 in the Dermatology Unit of San Donato Hospital of Arezzo and in the Section of Dermatology of University of Siena, Italy. Tumours varied from 0.8 to 4.4 cm in size. All patients underwent local excision; five also had lymph node dissection. One patient underwent adjuvant radiotherapy, and three received chemotherapy. RESULTS: Six of seven patients died, with survival varying from 15 to 45 months. Distant metastases occurred in two patients. Survival time was inversely proportional to the size of the tumour. CONCLUSIONS: MNH is an aggressive tumour and should be diagnosed and excised as early as possible. Histological parameters are paramount, but correct diagnosis also calls for attention to clinical presentation and any history of recurrence or recent enlargement of long-standing lesions. In our experience, radiotherapy and chemotherapy do not seem to prolong survival. PMID- 18410618 TI - Actinic keratosis: an occupational and environmental disorder. AB - Solar ultraviolet light electromagnetic waves are a known environmental carcinogenic agent closely associated with the development of skin cancer in light-complexioned individuals. Outdoor workers have higher annual exposure to ultraviolet light. We will review the topic of actinic keratoses among these individuals as this common rudimentary form of superficial cutaneous squamous cell carcinoma is explored in greater detail. PMID- 18410619 TI - Recall urticaria related to levofloxacin. PMID- 18410620 TI - Positivity of iron studies in pruritus of unknown origin. PMID- 18410621 TI - Surgical management of the eyelid and bulbar conjunctiva in a patient with nevus of Ota. PMID- 18410622 TI - A case of pretibial myxoedema associated to ectopic secreting thyroid nodule on thyroglossal duct residue. PMID- 18410623 TI - The role of immune serological parameters and allergological tests in psoriasis. PMID- 18410624 TI - Repigmentation of skin and hairs in stable vitiligo by transplantation of autologous melanocytes in fibrin suspension. PMID- 18410625 TI - Recurrent cuniculatum squamous cell carcinoma of the fingers and virus. PMID- 18410626 TI - Multiple and diffuse lipomas occurring after total body electron beam therapy for mycosis fungoides. PMID- 18410627 TI - Efficacy and tolerability of clindamycin phosphate and salicylic acid gel in the treatment of mild to moderate acne vulgaris. PMID- 18410628 TI - Successful therapy of a patient with therapy recalcitrant generalized bullous scleroderma by extracorporeal photopheresis and mycophenolate mofetil. PMID- 18410629 TI - Plantar exfoliation and desquamation during efalizumab therapy. PMID- 18410630 TI - Nevus depigmentosus affecting the iris and skin: a case report. PMID- 18410631 TI - Macular amyloidosis presenting in an incontinentia pigmenti-like pattern with subepidermal blister formation. PMID- 18410632 TI - Bcl-2 expression in dermal lymphocytes in lichen planus and psoriasis vulgaris. PMID- 18410635 TI - Special article: chronic granulomatous disease in the United Kingdom and Ireland: a comprehensive national patient-based registry. AB - There are no epidemiological studies from the British Isles of chronic granulomatous disease, characterized by recurrent, life-threatening bacterial and fungal infections and inflammatory sequelae. Patients were enrolled in a national registry and medical records were analysed. Of 94 subjects, 69 had X-linked disease, 16 had autosomal recessive disease and nine were unknown. Prevalence was 7.5/million for 1990-99 and 8.5/million for 1980-89. Suppurative adenitis, abscesses and pneumonia presented commonly. Twenty-three of 30 patients who underwent high resolution computerized tomography had chronic respiratory disease. Inflammatory sequelae included bowel stricture and urogenital tract granulomata. Growth failure was common; 75% of those measured were below the population mean. All patients received prophylactic antibiotics and 93% anti fungal prophylaxis. Interferon gamma was used to treat infection, but rarely as prophylaxis. Despite prophylaxis, estimated survival was 88% at 10 years but 55% at age 30 years. Morbidity remains significant, severe infectious complications common. Curative treatments including stem cell transplantation should be considered for patients with frequent or serious complications. PMID- 18410636 TI - Restoration of anti-tetanus toxoid responses in patients initiating highly active antiretroviral therapy with or without a boost immunization: an INITIO substudy. AB - INITIO is an open-labelled randomized trial evaluating first-line therapeutic strategies for human immunodeficiency virus-1 (HIV-1) infection. In an immunology substudy a tetanus toxoid booster (TTB) immunization was planned for 24 weeks after initiation of highly active antiretroviral therapy (HAART). All patients had received tetanus toxoid immunization in childhood. Generation of proliferative responses to tetanus toxoid was compared in two groups of patients, those receiving a protease inhibitor (PI)-sparing regimen (n = 21) and those receiving a PI-containing (n = 54) regimen. Fifty-two participants received a TTB immunization [PI-sparing (n = 15), PI-containing (n = 37)] and 23 participants did not [PI-sparing (n = 6) or PI-containing (n = 17)]. Cellular responses to tetanus antigen were monitored by lymphoproliferation at time of immunization and every 24 weeks to week 156. Proportions with a positive response (defined as stimulation index > or = 3 and Delta counts per minute > or = 3000) were compared at weeks 96 and 156. All analyses were intent-to-treat. Fifty-two participants had a TTB immunization at median 25 weeks; 23 patients did not. At weeks 96 and 156 there was no evidence of a difference in tetanus-specific responses, between those with or without TTB immunization (P = 0.2, P = 0.4). There was no difference in the proportion with response between those with PI-sparing or PI containing regimens at both time-points (P = 0.8, P = 0.7). The proliferative response to tetanus toxoid was unaffected by initial HAART regimen. Anti-tetanus responses appear to reconstitute eventually in most patients over 156 weeks when treated successfully with HAART, irrespective of whether or not a TTB immunization has been administered. PMID- 18410637 TI - Health behaviours, risk-taking and conceptual changes among schoolchildren aged 7 to 19 years in semi-rural Sweden. AB - BACKGROUND: This study covers a broad age group (7-19 years) and includes a wide range of themes. The aim is to describe how various behaviours, complaints and conceptual changes come into play and to discuss the factors that might support or hamper the happiness and well-being of growing children and adolescents. We also discuss the implications of our findings for future prevention programmes. METHODS: This cross-sectional study included all schoolchildren in a semi-rural district in Sweden (2181 pupils). A structured classroom questionnaire was used but the children were also given the opportunity to describe in their own words what was important for their happiness and well-being. RESULTS: Mean response rate was 85%. Most changes in behaviour occurred between 11 and 14 years of age. Girls had a more rapid process of change than boys. Both girls and boys experienced stress in their relations with peers, parents and teachers. Gender differences in emotional support were prominent. Regardless of age and sex, all schoolchildren asked for a richer choice of leisure time activities, a place where they could meet with friends and caring teachers, parents and adults in the surrounding community. CONCLUSIONS: Adolescence was perceived as a risky and problematic period not only by adults but also by the adolescents themselves. However, the perceived risks and the worries differed. While the adults mainly worried about the early onset of smoking and drug use, the schoolchildren worried about their social situation and their personal relationships. After decades of preventive programmes in Swedish schools, only modest results are seen. To be effective, future preventive programmes probably have to focus more on the conceptual world of the growing child. PMID- 18410638 TI - Risk factors for smoking among school adolescents in Greece. AB - OBJECTIVE: We conducted this study among school adolescents to identify factors, which influence schoolchildren to smoke. METHODS: We carried out a cross sectional study in a sample of 924 students of all classes (ages 12-18 years old) in 15 public high schools in a semi-urban area in Crete, Greece, using a questionnaire. The questionnaire comprised of 46 questions covering children's lifestyle habits regarding daily activities and leisure time, frequency of risk taking behaviour, knowledge about the hazards and long-term consequences of smoking. RESULTS: 23.9% of participants were experimental smokers and 18.6% were current smokers. 11.4% of the total population was daily smokers. There was a significant increase in the prevalence of experimental and current smokers with school grades, while peaks in last grades were observed. Boys started smoking earlier than girls, mean (standard error) age 13.4 (2.3) years vs. 14.1 (2.3) years, P = 0.01. Stepwise logistic regression analysis showed a positive relationship between current smoking and having brother or sister smoking [odds ratio (95% confidence interval) 2.7 (1.7-4.4) and 1.8 (1.1-3.3) respectively], having more than three friends who were smokers [2.6 (2-3.4)] and last school grade [1.4 (1.2-1.7)]. Students appeared to be informed about long-term smoking hazards and had negative views on children who smoke especially in the lower grades. CONCLUSIONS: Prevention programmes should be imposed early in elementary schools while cessation policies should target at all grades, in particular at critical grades depending on population-specific characteristics. PMID- 18410639 TI - The relationship between physical fitness and motor competence in children. AB - BACKGROUND: This study examined the relationship between physical fitness and motor competence in children aged 9-10 years. METHODS: A sample of 67 children (mean age 9.7, SD 0.3 year) participated in the study. To assess motor competence, each child completed the Movement Assessment Battery for Children. Physical fitness was measured by the Test of Physical Fitness, comprising nine different tasks including running, jumping, throwing and climbing. RESULTS: The results show a strong and significant correlation between motor competence and physical fitness within the sample. CONCLUSIONS: These results are relevant to the work of maintaining and developing both sufficient physical fitness and motor competence in children as these factors are important contributors to their health and well-being. PMID- 18410640 TI - 'Always the guiding hand': parents' accounts of the long-term implications of developmental co-ordination disorder for their children and families. AB - BACKGROUND: There has been little investigation of the long-term implications of developmental co-ordination disorder (DCD) and, particularly, its impact on families. This is despite the prevalence of the disorder (4.5% to 6%) of the child population, and the high rates of referral to occupational therapy departments. The study reported here was part of an evaluation of an innovative screening clinic for the assessment of children with DCD. METHODS: Within the case study approach of the evaluation, questionnaires, including open-end questions, were sent to 70 families. All had attended the screening clinic 6 years earlier and had little subsequent contact with the occupational therapy service. Semi-structured interviews were held with 12 mothers who volunteered to participate. These enabled specific issues raised in the questionnaires to be explored in greater depth. Interviews were audiotaped and full text transcripts produced for analysis. RESULTS: Over half the questionnaires were returned despite the length of time elapsed since hospital contact. Parents who responded reported a high persistence of problems in their children. Difficulties spanned motor and academic performance, emotional/behavioural responses and social interaction. Twenty-eight children (80%) of respondents were reported as having difficulties in three or more areas. Bullying was a commonly identified problem. At interview mothers spoke at length about their experiences and reported feeling stressed and distressed. Mothers reported a lack of support and expressed feelings of isolation. They said that their time investment in their child with DCD had pronounced effects on themselves and other family members. Specifically they highlighted time spent fighting the system, primarily for educational support. CONCLUSIONS: The study suggests a need for occupational therapists to reframe their current ideas regarding service provision, with improved support for families, increased interagency working and more service-user involvement. PMID- 18410641 TI - Healthcare needs and healthcare satisfaction from the perspective of parents of children with chronic conditions: the DISABKIDS approach towards instrument development. AB - AIM: Increasingly, families' perspectives are taken into account in the appraisal of health services. The objective of this study was to cross-culturally analyse concepts related to healthcare needs, healthcare utilization and the appraisal and satisfaction with care of parents of children with chronic conditions with the aim of developing a cross-cultural measure. METHODS: Several approaches were employed in the study: (i) a deductive approach integrating existing measurements; and (ii) an inductive approach based on focus groups. Focus groups were conducted in seven countries with mothers and fathers as well as their children with seven different chronic conditions, and qualitatively analysed. RESULTS: As a result of an evaluation of the different existing methodological approaches, the basic structural components were identified: healthcare needs, the receipt of services, problems with receiving services as well as the appraisal of and satisfaction with the quality of care. While items referring to existing healthcare services were primarily derived by the work of an expert group, items related to quality of care and satisfaction with services mainly evolved from the focus group work. From the focus groups, 367 statements were extracted, which were further processed in a Q-sort rating by a multinational expert group in order to identify domains and salient items. The draft questionnaire to be pilot tested cross-nationally consisted of 101 items which were reduced on the basis of psychometric findings. CONCLUSION: On the basis of results of focus groups and existing evidence, a comprehensive measure should be employed in paediatric health services research including structural, process and outcome parameters of care from the perspective of parents. PMID- 18410642 TI - Family-centred service coordination in childhood health and disability services: the search for meaningful service outcome measures. AB - BACKGROUND: Potential service outcome measures were tested for their utility in the assessment of the quality of 'family centred' service coordination in the provincial network of children's disability services in Manitoba, Canada. METHODS: This study is based on in-home survey data provided by 103 mothers at 6 and 18 months following assignment of a 'dedicated' service coordinator. Service outcome indicators included measures of parent self-esteem, parenting stress, family functioning and the need for family support resources. RESULTS: Hierarchical regression analyses showed no relationship between level of quality of family-centred service coordination and standardized psychosocial measures of parent and family functioning. However, family centredness of service coordination was found to predict significant reduction in level of family need for psychosocial support resources after 18 months of contact with a service coordinator. CONCLUSIONS: Outcome measures that are focused on specific and tangible results of service coordination appear to be of higher utility in service quality assessment than are more global, standardized measures of parent and family functioning. PMID- 18410643 TI - Included but isolated: early intervention programmes provision for children and families with chronic respiratory support needs. AB - PURPOSE: To measure enrollment of children with mechanical respiratory support needs within the Massachusetts early intervention programmes (EIP) and describe challenges in execution of individual family service plans (IFSPs). METHODS: Mixed methods provided a complementary assessment. Quantitative data were collected in 2005 from the Commonwealth of Massachusetts EIP administrative database as part of a cross-sectional state census. Qualitative data were retrieved from EIP regional coordinators in a key informant focus group. Descriptive statistics were used for quantitative survey data. Audio-recordings were transcribed verbatim and a qualitative, thematic analysis was undertaken. RESULTS: Four hundred and eighty children requiring mechanical respiratory support at the time of EIP enrollment were identified between 1 July 1997 and 1 January 2005. Focus group analysis revealed themes including: (1) barriers to community transition; (2) community expertise and training; (3) interface with medical providers; and (4) the role of web-based resources. Isolation of families emerged as a recurrent and resounding concept, relating to all of the themes. CONCLUSIONS: Findings support the assumption that implementation of IFSPs for the cohort of children with chronic mechanical respiratory support needs in EIPs is challenging. Barriers reflect inconsistent care coordination and practical encumbrances, contributing to the physical and social isolation of these children and their families. PMID- 18410644 TI - Efficacy of the Incredible Years Programme as an early intervention for children with conduct problems and ADHD: long-term follow-up. AB - BACKGROUND: This study examined the long-term efficacy of the Incredible Years (IY) BASIC Parenting Programme delivered as a preventive intervention with parents of pre-school children who display signs of attention deficit hyperactivity disorder (ADHD) and conduct problems. Families were followed up after the completion of a controlled trial with 11 Sure Start areas in North and Mid-Wales and North West England. METHODS: Participants in the study were 50 pre school children whose parents had received the intervention. Child ADHD symptoms were assessed at baseline, at follow-up one (6 months after baseline); at follow up two (12 months after baseline); and at follow-up three (18 months after baseline). Families in the original waiting-list control group were not assessed after follow-up one as they had subsequently received the same intervention. RESULTS: The significant post-intervention improvements in child ADHD symptoms evident at follow-up one were maintained over time, as demonstrated by statistical and clinical stability of measures. No significant differences were found for ADHD symptoms across each follow-up, indicating that the gains made post intervention were maintained for at least 12 months, with 57% of the sample maintaining scores below the clinical cut-off on the Conners. Eighty-six, 58, and 30 per cent respectively had maintained at least a modest, large, or very large improvement in ADHD symptoms at follow-up three. CONCLUSIONS: Findings from this study suggest that the IY psychosocial treatment programme is a valuable intervention in the longer term for many pre-school children displaying early signs of ADHD. PMID- 18410645 TI - The Leader Observation Tool: a process skills treatment fidelity measure for the Incredible Years parenting programme. AB - BACKGROUND: Despite recognition of the need to deliver evidence-based programmes in the field of mental health, there is little emphasis on implementing such programmes with fidelity. Attempts by programme developers to ensure adherence to their programmes include the development of training, manuals and content scales, but these alone may be insufficient to ensure fidelity in replication. Observational measures lend themselves as a potentially useful assessment of intervention outcomes, providing accurate and objective accounts of the intervention process. AIM: To develop a reliable and valid observational treatment fidelity tool of process skills required to deliver the Incredible Years (IY) BASIC parenting programme effectively. METHODS: An objective observational fidelity measure was developed to assess adherence to the IY BASIC parenting programme protocol. Observations were conducted on 12 IY BASIC parenting programme groups, attended by parents of pre-school children displaying signs of early onset conduct disorder. RESULTS: The Leader Observation Tool (LOT) achieved high internal reliability and good code-recode and inter-rater reliability. Evidence of concurrent validity was also obtained. CONCLUSIONS: Having demonstrated that the LOT is a reliable and valid measure of implementation fidelity, further research is necessary to examine the relationship between LOT scores and intervention outcome. PMID- 18410646 TI - Transitional care for adolescents with rheumatic diseases: urgent need for more research. PMID- 18410649 TI - Exploring the mechanisms of intrapartum transmission of HIV. Does elective caesarean section hold the key? PMID- 18410650 TI - Maternal depression and medication exposure during pregnancy: comparison of maternal retrospective recall to prospective documentation. AB - OBJECTIVE: Outcome investigations of prenatal maternal depression and psychotropic exposure rely extensively on maternal retrospective recall. This study compared postnatal recall to prospective documentation of illness and medication exposures. DESIGN: Prospective cohort and retrospective case-control studies. SETTING: Emory Women's Mental Health Program (prospective study) and Emory University Department of Psychology (retrospective study). SAMPLE: A total of 164 women who participated in both the prospective and retrospective studies. METHODS: Women with a history of mental illness were followed during pregnancy for prospective prenatal assessments of depression and medication exposures. At 6 months postpartum, some of these women also participated in a retrospective study during which they were asked to recall prenatal depression and medication use. Agreement between prospective and retrospective documentation of exposures was analysed. MAIN OUTCOME MEASURES: Occurrence of maternal depression during pregnancy and maternal use of pharmacological agents during pregnancy. RESULTS: There was only moderate agreement (k = 0.42) in prospective versus retrospective reporting of prenatal depression. Positive predictive value for recalling depression was 90.4%; however, negative predictive value for denying depression was only 53.8%. Participants accurately recalled psychotropic use but significantly underreported use of nonpsychotropic medications. CONCLUSIONS: Studies using retrospective data collection may be susceptible to systematic recall bias with underreporting of maternal depression and use of nonpsychotropic agents during pregnancy. PMID- 18410651 TI - Survey of prenatal screening policies in Europe for structural malformations and chromosome anomalies, and their impact on detection and termination rates for neural tube defects and Down's syndrome. AB - OBJECTIVE: To 'map' the current (2004) state of prenatal screening in Europe. DESIGN: (i) Survey of country policies and (ii) analysis of data from EUROCAT (European Surveillance of Congenital Anomalies) population-based congenital anomaly registers. SETTING: Europe. POPULATION: Survey of prenatal screening policies in 18 countries and 1.13 million births in 12 countries in 2002-04. METHODS: (i) Questionnaire on national screening policies and termination of pregnancy for fetal anomaly (TOPFA) laws in 2004. (ii) Analysis of data on prenatal detection and termination for Down's syndrome and neural tube defects (NTDs) using the EUROCAT database. MAIN OUTCOME MEASURES: Existence of national prenatal screening policies, legal gestation limit for TOPFA, prenatal detection and termination rates for Down's syndrome and NTD. RESULTS: Ten of the 18 countries had a national country-wide policy for Down's syndrome screening and 14/18 for structural anomaly scanning. Sixty-eight percent of Down's syndrome cases (range 0-95%) were detected prenatally, of which 88% resulted in termination of pregnancy. Eighty-eight percent (range 25-94%) of cases of NTD were prenatally detected, of which 88% resulted in termination. Countries with a first-trimester screening policy had the highest proportion of prenatally diagnosed Down's syndrome cases. Countries with no official national Down's syndrome screening or structural anomaly scan policy had the lowest proportion of prenatally diagnosed Down's syndrome and NTD cases. Six of the 18 countries had a legal gestational age limit for TOPFA, and in two countries, termination of pregnancy was illegal at any gestation. CONCLUSIONS: There are large differences in screening policies between countries in Europe. These, as well as organisational and cultural factors, are associated with wide country variation in prenatal detection rates for Down's syndrome and NTD. PMID- 18410652 TI - Delayed umbilical cord clamping at birth has effects on arterial and venous blood gases and lactate concentrations. AB - OBJECTIVE: To estimate the influence of delayed umbilical cord clamping at birth on arterial and venous umbilical cord blood gases, bicarbonate (HCO3-), base excess (BE) and lactate in vigorous newborns. SETTING: University hospital. DESIGN: Prospective observational. SAMPLE: Vaginally delivered term newborns. MATERIAL AND METHODS: Umbilical cord arterial and venous blood was sampled repeatedly every 45 seconds (T(0)= time zero; T(45)= 45 seconds, T(90)= 90 seconds) until the cord pulsations spontaneously ceased in 66 vigorous singletons with cephalic vaginal delivery at 36-42 weeks. Longitudinal comparisons were performed with the Wilcoxon signed-ranks matched pairs test. Mixed effect models were used to describe the shape of the regression curves. MAIN OUTCOME MEASURES: Longitudinal changes of umbilical cord blood gases and lactate. RESULTS: In arterial cord blood, there were significant decreases of pH (7.24-7.21), HCO3- (18.9-18.1 mmol/l) and BE (-4.85 to -6.14 mmol/l), and significant increases of PaCO(2) (7.64-8.07 kPa), PO(2) (2.30-2.74 kPa) and lactate (5.3-5.9 mmol/l) from T(0) to T(90), with the most pronounced changes at T(0)-T(45). Similar changes occurred in venous blood pH (7.32-7.31), HCO3- (19.54-19.33 mmol/l), BE (-4.93 to -5.19 mmol/l), PaCO(2) (5.69-5.81 kPa) and lactate (5.0-5.3 mmol/l), although the changes were smaller and most pronounced at T(45)-T(90). No significant changes were observed in venous PO(2). CONCLUSION: Persistent cord pulsations and delayed cord clamping at birth result in significantly different measured values of cord blood acid-base parameters. PMID- 18410653 TI - Lactate concentration in umbilical cord blood is gestational age-dependent: a population-based study of 17 867 newborns. AB - OBJECTIVE: To study the influence of gestational age on lactate concentration in arterial and venous umbilical cord blood at birth and to define gestational age specific reference values for lactate in vigorous newborns. DESIGN: Population based comparative. SETTING: University hospitals. SAMPLE: Vigorous newborns with validated umbilical cord blood samples. MATERIAL AND METHODS: From 2000 to 2004, routine cord blood gases, lactate and obstetric data from two university hospitals were available for 17 867 newborns from gestational week 24 to 43. After validation of blood samples and inclusion only of singleton pregnancies aimed for vaginal delivery, 10 700 women remained. Among those, reference values were defined in 10 169 vigorous newborns, that is in newborns with a 5-minute Apgar score corresponding to the gestational age-specific median value minus 1 point score, or better. MAIN OUTCOME MEASURES: Cord lactate concentration relative to gestational age. RESULTS: The arterial and venous lactate concentrations increased monotonously with gestational age from 34 weeks. Considerable differences were found between mean and median values, but after logarithmic transformation the log-lactate values were normally distributed. Simple linear regression analysis showed a significant association between the log-lactate values and gestational age (P < 10(-6), R(2)= 0.024). Reference curves were constructed after anti-logarithmic transformation. Both the gestational age and the time of the second stage of labour influenced, independently of each other, the lactate concentrations. CONCLUSIONS: Lactate concentrations in arterial and venous umbilical cord blood are increasing significantly with advancing gestational age. PMID- 18410654 TI - Ethnic differences in preterm birth and its subtypes: the effect of a cumulative risk profile. AB - OBJECTIVE: To explore the effect of potentially explanatory risk factors on ethnic differences in the prevalence of preterm birth (PTB) and its subtypes. DESIGN: Prospective population-based cohort study. SETTING AND POPULATION: Pregnant women from Amsterdam attending their first antenatal visit for obstetric care. A total of 8266 women participated (response rate 67%). Ethnicity was based on the country of birth of the pregnant woman's mother: the Netherlands, Surinam, the Antilles, Turkey, Morocco, Ghana and other non-Dutch countries. Exclusion criteria were multiple births and gestational age at delivery less than 24 weeks. METHODS: Risk factors were obtained using a multilingual questionnaire and from the Dutch Perinatal Registration. Risk factors were summed into a cumulative risk score. Multiple logistic regression analyses were performed. MAIN OUTCOME MEASURES: Odds ratios with 95% CIs were calculated for total, spontaneous and iatrogenic (medically indicated) preterm births for the ethnic minority groups versus the Dutch reference group. RESULTS: After adjustment for all risk factors, the Surinamese (OR 1.6, 95% CI 1.2-2.4), Ghanaian (OR 2.0, 95% CI 1.1-3.6) and Antillean (OR 1.6, 95% CI 0.8-3.3) women had a higher risk of PTB compared with the Dutch women, in particular for iatrogenic preterm birth (OR 2.1, 95% CI 1.0 4.4; OR 3.2, 95% CI 1.0-10.4; OR 3.6, 95% CI 1.1-11.2, respectively). The ethnic minority groups had a higher cumulative risk score (ranging from 2.1 to 3.7) compared with the Dutch group (1.8). Adjustment for the cumulative risk score considerably decreased the risk of PTB among the Surinamese (OR 1.2, 95% CI 0.9 1.7), Ghanaian (OR 1.3, 95% CI 0.8-2.3) and Antillean (OR 1.2, 95% CI 0.6-2.4) women. CONCLUSIONS: A cumulation of risk factors, mainly observed among the ethnic minority groups, contributes to the explanation of ethnic differences in PTB prevalence. PMID- 18410655 TI - Maternal obesity, length of gestation, risk of postdates pregnancy and spontaneous onset of labour at term. AB - OBJECTIVE: To investigate the effect of maternal body mass index (BMI) on postdates pregnancy, length of gestation and likelihood of spontaneous onset of labour at term. DESIGN: Retrospective cohort study. SETTING: Swedish Medical Birth Register. POPULATION: A total of 186 087 primiparous women (of whom 143 519 had spontaneous onset of labour at term) who gave birth between 1998 and 2002. METHODS: Mann-Whitney test, one-way analysis of variance, linear regression and single variable logistic regression. MAIN OUTCOME MEASURES: Postdates pregnancy (>/=294 days or 42(+0) weeks), length of gestation and likelihood of spontaneous onset of labour at term. RESULTS: About 6.8% of pregnancies delivered postdates. Higher maternal BMI (kg/m(2)) during the first trimester was associated with longer gestation (P < 0.001) as was a greater change in BMI between the first and third trimesters (BMI measured on admission prior to delivery) with mean (SD) gestation at delivery of 280.7 (8.6) and 283.2 (8.6) days for increases in BMI of <2 and >/=10 kg/m(2), respectively. Higher BMI during the first trimester was associated with a lower chance of spontaneous onset of labour at term. Compared with BMI 20 to <25 kg/m(2), the odds ratios (95% CI) for spontaneous onset of labour at term were 1.21 (1.15-1.27) for BMI of <20 kg/m(2), 0.71 (0.69-0.74) for BMI of 25 to <30 kg/m(2), 0.57 (0.54-0.60) for BMI of 30 to <35 kg/m(2) and 0.43 (0.40-0.47) for BMI of >/=35 kg/m(2). Higher BMI during the first trimester (BMI of >/=35 kg/m(2) compared with BMI of 20 to <25 kg/m(2)) was also associated with an increased risk of complications including stillbirth (OR 3.90, 95% CI 2.44 6.22), gestational diabetes (OR 5.61, 95% CI 4.61-6.83) and caesarean section (OR 2.39; 95% CI 2.20-2.59). CONCLUSIONS: Higher maternal BMI in the first trimester and a greater change in BMI during pregnancy were associated with longer gestation and an increased risk of postdates pregnancy. Higher maternal BMI during the first trimester was also associated with decreased likelihood of spontaneous onset of labour at term and increased likelihood of complications. PMID- 18410656 TI - The risk of unexplained antepartum stillbirth in second pregnancies following caesarean section in the first pregnancy. AB - OBJECTIVE: To determine if a previous caesarean section increases the risk of unexplained antepartum stillbirth in second pregnancies. STUDY DESIGN: Retrospective cohort study. SETTING: Large Canadian perinatal database. POPULATION: 158 502 second births. METHODS: Data were obtained from a large perinatal database, which supplied data on demographics, pregnancy complications, maternal medical conditions, previous caesarean section and pregnancy outcomes. MAIN OUTCOME MEASURES: Total and unexplained stillbirth. RESULTS: The antepartum stillbirth rate was 3.0/1000 in the previous caesarean section group compared with 2.7/1000 in the previous vaginal delivery group (P= 0.46). Multivariate logistic regression modelling, including terms for maternal age (polynomial), weight >91 kg, smoking during pregnancy, pre-pregnancy hypertension and diabetes, did not document an association between previous caesarean section and unexplained antepartum stillbirth (OR 1.27, 95% CI 0.92-1.77). CONCLUSION: Caesarean section in the first birth does not increase the risk of unexplained antepartum stillbirth in second pregnancies. PMID- 18410657 TI - Substandard care in maternal mortality due to hypertensive disease in pregnancy in the Netherlands. AB - OBJECTIVES: To review the standard of care in cases of maternal mortality due to hypertensive diseases in pregnancy and to make recommendations for its improvement. DESIGN: Care given to women with hypertensive disease in pregnancy was audited and substandard care factors identified. SETTING: Confidential enquiry by the Dutch Maternal Mortality Committee (MMC) from the Netherlands Society of Obstetrics and Gynaecology. POPULATION: All maternal deaths reported to the MMC due to hypertensive disease in pregnancy in the Netherlands during the years 2000-04. METHODS: Assessment for substandard care factors using a checklist based on the Dutch guideline of 'Hypertensive Disorders in Pregnancy'. MAIN OUTCOME MEASURES: Substandard care in cases of maternal mortality due to hypertensive diseases in pregnancy. RESULTS: A total of 27 cases of maternal death due to hypertensive disease in pregnancy were reported to the committee in the study period. In 26 cases (96%), substandard care factors were present, of which in 17 cases (63%), these were for more than five different items. In community midwifery care, the most frequent substandard care factor was no testing for proteinuria when clearly indicated (41%). In hospital care, the most frequent substandard care was related to insufficient diagnostic testing when indicated (41%), insufficient management of hypertension by obstetricians (85%), no use or inadequate use of magnesium sulphate (67%), inadequate stabilisation before transport to tertiary care centres and/or delivery (52%) and failure to consider timely delivery (44%). CONCLUSIONS: Education of pregnant women concerning danger signs of hypertensive disease should be improved. Training of midwives and obstetricians should be improved in the following areas: performing basic diagnostic tests, adequate management of hypertension and eclampsia, with more attention to treatment of systolic blood pressure. This training should be guided by clear local protocols. Delivery should not be delayed in serious cases of hypertensive disease in pregnancy, not only after 32-34 weeks but also in early-onset pre-eclampsia as maternal risks often outweigh possible fetal benefits of temporising management. PMID- 18410658 TI - Swyer syndrome: presentation and outcomes. AB - OBJECTIVE: To establish the spectrum of presentation, natural history and gynaecological outcomes in women with Swyer syndrome. DESIGN: Retrospective notes review. SETTING: Tertiary referral centre for disorders of sex development. POPULATION: A total of 29 adult women with Swyer syndrome. METHODS: Information was collected on age at diagnosis, biometric characteristics, timing of gonadectomy, histology of gonad, bone mineral density, uterine size and fertility. MAIN OUTCOME MEASURES: Age at diagnosis, risk of gonadal malignancy, bone mineral density, uterine size. RESULTS: With regard to presentation, 26/29 (90%) women in this series presented with delayed puberty, and the median age at diagnosis was 17.2 years (range 0-55 years). The median age at gonadectomy was 18 years (range 9-33 years). Histology of the gonad was available in 22 women and demonstrated streak gonads with no evidence of malignancy in 12, dysgerminoma in 7 and gonadoblastoma in 3. The youngest patient diagnosed with dysgerminoma was 10 years old. The median height of the women was 1.73 m (range 1.54-1.95 m). Twelve out of the 20 (60%) women had evidence of osteopenia on dual energy X-ray absorptiometry scan. The uterine size and shape was assessed in eight women after completion of induction of puberty, and the uterine cross-section was found to be significantly lower than that in normal controls. Fertility was achieved with ovum donation in three women, all of whom had live births and one subsequently had a second successful pregnancy. CONCLUSION: Early diagnosis of Swyer syndrome is necessary in view of the risk of dysgerminoma that can develop at an early age. Adequate hormone replacement is required to maintain bone mineral density and may improve the uterine size and shape. PMID- 18410659 TI - Efficacy of laparoscopic subtotal hysterectomy in the management of menorrhagia: 400 consecutive cases. AB - STUDY OBJECTIVE: To assess the safety and patient satisfaction of laparoscopic subtotal hysterectomy (LSH) using a standardised surgical technique. DESIGN: Prospective observational study. SETTING: Princess Royal University Hospital, Chelsfield Park Hospital and Sloane Hospital, Kent, UK. PATIENTS AND MATERIALS: Four hundred consecutive women with menorrhagia underwent LSH. The procedure was performed using the Plasma Kinetic Bipolar Diathermy (Gyrus International Ltd, Berkshire, UK) for pedicle ligation and the Lap Loop system (Roberts Surgical Healthcare Ltd, Kidderminster, UK) to detach the cervix. An electromechanical morcellator (Morcellex; Ethicon Women's Health and Urology, Cincinnati, OH, USA) was used to remove the uterus from the abdominal cavity. MAIN OUTCOME MEASURES: Patient satisfaction, morbidity rates and readmission rates. RESULTS: A total of 400 LSH were performed between February 2003 and November 2006. The principal clinical indication for hysterectomy was menorrhagia. The mean duration of surgery was 46.4 minutes. The mean operative blood loss was 126 ml. Concurrent surgery was performed in 141 women. Minor and major perioperative complications were encountered in 5% (n= 20) of women. The major complication rate was 1.2% (n= 5): three women (0.75%) with bladder perforation, two women (0.5%) with bowel injury and one woman (0.25%) with a vesicocervical fistula. Eight women (2%) suffered from cyclical vaginal bleeding postoperatively. CONCLUSIONS: LSH is a safe and effective treatment for menorrhagia and other menstrual disorders when hysterectomy is indicated. Women appreciate the quick recovery period, reduced time off work and faster return to normal activity. Our data suggest that LSH can replace abdominal hysterectomy in selected cases. PMID- 18410660 TI - Modelling the cost-effectiveness and capacity impact of changes to colposcopy referral guidelines for women with mild dyskaryosis in the UK Cervical Screening Programme. AB - OBJECTIVE: To evaluate the capacity implications and health economic impact of new guidelines recommending referral to colposcopy after one mild result during cervical screening rather than after two consecutive mild results. DESIGN: A mathematical model of the country's colposcopy services and the clinical pathways from smear result through to treatment is constructed. The model incorporates national questionnaire data on referral numbers and management practices, routine data and published research results. SETTING: All English NHS colposcopy services. POPULATION: Women aged 25 to 64 years. METHODS: The national average workload impact of the change in referral guidelines is predicted, and the impact in differing local circumstances is evaluated within the model. A long-term health economic model examines the resulting costs and predicted change in quality-adjusted life years (QALYs). MAIN OUTCOME MEASURES: Colposcopy workload implications for single mild dyskaryosis referral and cost per QALY analysis. RESULTS: We found that single mild dyskaryosis referral implies, on average, a 21% increase in colposcopy workload for services not currently operating this policy. The health economic model predicted a cost per QALY gained as a result of the implementation of the new referral guidelines of around pound7,500. CONCLUSIONS: Referral after one mild result will increase workload at colposcopy; however, it may be possible to counterbalance the additional workload by altering other clinical practice. The change to referral guidelines would be considered cost-effective in comparison with many interventions routinely available on the NHS. PMID- 18410661 TI - Inferior vena cava filter use in pregnancy: preliminary experience. AB - Thromboembolism is the most common direct cause of maternal mortality in the UK. Inferior vena cava (IVC) filter placement is indicated in conditions where recurrent thromboembolism occurs despite adequate anticoagulation or when anticoagulation is contraindicated. The safety of IVC filter use in pregnancy is uncertain, as there are limited data available. In this study, we have reviewed pregnancy outcome in women with IVC filter use. Twelve pregnancies in six women, delivered in our hospital in the past 11 years, were identified from obstetric and radiology databases to have an IVC filter in situ. In four pregnancies, an IVC filter was placed during pregnancy. In eight pregnancies, an IVC filter was already in situ before pregnancy and continued for the entire duration of pregnancy. There were no antenatal complications noted due to IVC filter placement and no recurrent thromboembolism noted in pregnancies with an IVC filter in situ before conception. The mode of delivery was based on obstetric reasons in all cases. The mean birthweight was 2982 g, and all babies were born in good condition with Apgar scores within normal range. In conclusion, this case series did not identify any problems associated with IVC filter placement or continuation in pregnancy. PMID- 18410662 TI - Massive postpartum haemorrhage after uterus-conserving surgery in placenta percreta: the danger of the partial placenta percreta. AB - Placenta percreta is a rare but potentially life-threatening condition associated with high maternal mortality and morbidity rates, usually arising from severe obstetric haemorrhage. Due to rising caesarean section rates, an increase in the incidence of morbidly adherent placentas (accreta, increta and percreta) has been observed. Various treatment strategies have been employed in different centres, ranging from performing a caesarean hysterectomy at the time of delivery to leaving the placenta in situ, with or without adjuvant internal iliac and uterine arterial embolisation and/or methotrexate therapy. In the case of placenta percreta, irrespective of the treatment method employed, women are still at high risk of life-threatening haemorrhage and morbidity secondary to placental invasion beyond the confines of the uterine serosa into surrounding organs, most commonly the bladder. We describe an unusual case of a partially adherent placenta percreta in which partial separation of the normally implanted placenta led to torrential haemorrhage on the third postoperative day after the placenta was left in situ at the time of delivery. We therefore advise caution in following a conservative approach in the treatment of cases of placenta percreta in which the percreta feature is only partial and will discuss the merits and disadvantages of alternative options. PMID- 18410663 TI - Weight gain restriction for obese pregnant women: a case-control intervention study. PMID- 18410666 TI - Increased perinatal mortality and morbidity in monochorionic versus dichorionic twin pregnancies: clinical implications of a large Dutch cohort study. PMID- 18410668 TI - Cervical stitch (cerclage) for preventing pregnancy loss: individual patient data meta-analysis. PMID- 18410669 TI - Peripartum hysterectomy in the UK: management and outcomes of the associated haemorrhage. PMID- 18410671 TI - Errors and misquotations in study on narghile and newborn birthweight. PMID- 18410674 TI - Cross-sectional prospective survey to study indication-based usage of antimicrobials in animals: results of use in cattle. AB - BACKGROUND: Indication-based data on the use of antimicrobials in animals were collected using a prospective cross-sectional survey, similarly as for surveys carried out in human medicine, but adapting the questionnaire to include veterinary-specific issues. The participating veterinarians were randomly selected from a sample population of practising veterinarians. The sampling was stratified to take into account the proportions of different types of veterinary practice in the country. All patients consulting the veterinary practice during a 1-week period were included in the study and veterinarians returned a completed questionnaire for each patient receiving antimicrobial treatment. As cattle received most of the treatments, results from the survey are given using cattle as an example species. RESULTS: The survey was sent to 681 veterinarians, of whom 262 (39%) responded. In total 2850 questionnaires were completed. The largest quantities of antimicrobials, measured in kilograms, were used for cattle, followed by pigs, dogs and horses. The species that were treated most were cattle (n = 1308), dogs (n = 989) and cats (n = 311). For cattle, the most common reason for treatment was acute mastitis (52%), followed by dry-cow therapy (21%), subclinical mastitis (6%) and treatment for acute enteritis (4%). The remaining treatments covered 17% of cattle patients and 15 different indications. For acute mastitis, parenteral or intramammary treatment was used in 36% and 34% of the cases, respectively. The remaining 30% received both treatments simultaneously. Of the parenteral treatments (n = 459), benzyl penicillin was used in 83% of the treated animals (n = 379), while fluoroquinolones were used in 49 cases (11%). Of the 433 cows receiving intramammary treatment, ampicillin combined with cloxacillin was most commonly used (n = 157; 36%), followed by cephalexin+streptomycin (n = 113; 26%). CONCLUSION: This cross-sectional prospective survey provided a useful method for the collection of information on the indication-based use of antimicrobials in different animal species. Cattle were the most commonly treated animal species during the study period. The most common indication for antimicrobial use in cows was mastitis. Benzyl penicillin was the drug most frequently used for the treatment of mastitis, which seems appropriate according to the national guidelines on the use of antimicrobials in cattle in Finland. PMID- 18410675 TI - ReXSpecies--a tool for the analysis of the evolution of gene regulation across species. AB - BACKGROUND: Annotated phylogenetic trees that display the evolution of transcription factor binding in regulatory regions are useful for e.g. 1) narrowing down true positive predicted binding sites, providing predictions for binding sites that can be tested experimentally, and 2) giving insight into the evolution of gene regulation and regulatory networks. RESULTS: We describe ReXSpecies, a web-server that processes the sequence information of a regulatory region for multiple species and associated (predicted) transcription factor binding sites into two figures: a) An annotated alignment of sequence and binding sites, consolidated and filtered for ease of use, and b) an annotated tree labeled by the gain and loss of binding sites, where the tree can be calculated from the data or taken from a trusted taxonomy, and the labels are calculated based on standard or Dollo parsimony. For genes involved in mammalian pluripotency, ReXSpecies trees highlight useful patterns of transcription factor binding site gain and loss, e.g. for the Oct and Sox group of factors in the 3' untranslated region of the cystic fibrosis transmembrane conductance regulator gene, which closely match experimental data. CONCLUSION: ReXSpecies post processes the information provided by transcription factor binding site prediction tools, in order to compare data from many species. The tool eases visualization and successive interpretation of transcription factor binding data in an evolutionary context. The ReXSpecies URL can be found in the Availability and requirements section. PMID- 18410676 TI - Whole genome comparative studies between chicken and turkey and their implications for avian genome evolution. AB - BACKGROUND: Comparative genomics is a powerful means of establishing inter specific relationships between gene function/location and allows insight into genomic rearrangements, conservation and evolutionary phylogeny. The availability of the complete sequence of the chicken genome has initiated the development of detailed genomic information in other birds including turkey, an agriculturally important species where mapping has hitherto focused on linkage with limited physical information. No molecular study has yet examined conservation of avian microchromosomes, nor differences in copy number variants (CNVs) between birds. RESULTS: We present a detailed comparative cytogenetic map between chicken and turkey based on reciprocal chromosome painting and mapping of 338 chicken BACs to turkey metaphases. Two inter-chromosomal changes (both involving centromeres) and three pericentric inversions have been identified between chicken and turkey; and array CGH identified 16 inter-specific CNVs. CONCLUSION: This is the first study to combine the modalities of zoo-FISH and array CGH between different avian species. The first insight into the conservation of microchromosomes, the first comparative cytogenetic map of any bird and the first appraisal of CNVs between birds is provided. Results suggest that avian genomes have remained relatively stable during evolution compared to mammalian equivalents. PMID- 18410677 TI - The influence of multi-morbidity and self-reported socio-economic standing on the prevalence of depression in an elderly Hong Kong population. AB - BACKGROUND: There has been an increasing prevalence of both depression and chronic medical conditions globally but the relationship between depression and multi-morbidity is not well understood. The aim of the present study was to investigate the relationship between depression, multi-morbidity (number of chronic medical conditions, and measures of socioeconomic standing (SES) in an elderly Hong Kong population. METHODS: Cross sectional study. Information on clinically relevant depressive symptoms, measured by the Geriatric Depression Scale (GDS), and demographic and chronic medical conditions were collected using standardized questionnaires. Information collected on SES included educational status (ES), maximum ever income (MEI), and self-perceived social standing in local community (SES-COM) and in Hong Kong generally (SES-HK). Analysis was conducted using multiple logistic regression RESULTS: Depression rates were similar in men and women (GDS caseness 8.1% vs 8.4%). Multi-morbidity of chronic medical conditions was common (40% of men and 46% of women had three or more). In the overall sample, the prevalence of depression was associated with the number of chronic medical conditions (OR 1.27; CI: 1.16-1.39). In addition, SES-HK and SES-COM were significant independent variables. CONCLUSION: In this elderly Hong Kong population, depression prevalence rose markedly with number of chronic medical conditions and SES-HK and SES-COM. PMID- 18410678 TI - Natural language processing in aid of FlyBase curators. AB - BACKGROUND: Despite increasing interest in applying Natural Language Processing (NLP) to biomedical text, whether this technology can facilitate tasks such as database curation remains unclear. RESULTS: PaperBrowser is the first NLP-powered interface that was developed under a user-centered approach to improve the way in which FlyBase curators navigate an article. In this paper, we first discuss how observing curators at work informed the design and evaluation of PaperBrowser. Then, we present how we appraise PaperBrowser's navigational functionalities in a user-based study using a text highlighting task and evaluation criteria of Human Computer Interaction. Our results show that PaperBrowser reduces the amount of interactions between two highlighting events and therefore improves navigational efficiency by about 58% compared to the navigational mechanism that was previously available to the curators. Moreover, PaperBrowser is shown to provide curators with enhanced navigational utility by over 74% irrespective of the different ways in which they highlight text in the article. CONCLUSION: We show that state-of-the-art performance in certain NLP tasks such as Named Entity Recognition and Anaphora Resolution can be combined with the navigational functionalities of PaperBrowser to support curation quite successfully. PMID- 18410679 TI - Spectrophotometric determination of etodolac in pure form and pharmaceutical formulations. AB - BACKGROUND: Etodolac (ETD) is a non-steroidal anti-inflamatory antirheumatic drug. A survey of the literature reveals that there is no method available for the determination of ETD in pure form and pharmaceutical formulations by oxidation-reduction reactions. RESULTS: We describe three simple, sensitive and reproducible spectrophotometric assays (A-C) for the determination of etodolac in pure form and in pharmaceutical formulations. Methods A and B are based on the oxidation of etodolac by Fe3+ in the presence of o-phenanthroline (o-phen) or bipyridyl (bipy). The formation of the tris-complex on reaction with Fe3+-o-phen and/or Fe3+-bipy mixtures in acetate buffer solution at optimum pH was demonstrated at 510 and 520 nm with o-phen and bipy. Method C is based on the oxidation of etodolac by Fe3+ in acidic medium, and the subsequent interaction of iron(II) with ferricyanide to form Prussian blue, with the product exhibiting an absorption maximum at 726 nm. The concentration ranges are 0.5-8, 1.0-10 and 2-18 microg mL(-1) respectively for methods A, B and C. For more accurate analysis, Ringbom optimum concentration ranges were calculated, in addition to molar absorptivity, Sandell sensitivity, detection and quantification limits. CONCLUSION: Our methods were successfully applied to the determination of etodolac in bulk and pharmaceutical formulations without any interference from common excipients. The relative standard deviations were or =10; 66.7% of NoV outbreaks occurred in restaurants; no differences in the attack rates were observed according to the etiology. Hospitalizations were more common (p = 0.03) in bacterial outbreaks (8.6%) than in NoV outbreaks (0.15%). Secondary cases accounted for 4% of cases in NoV outbreaks compared with 0.3% of cases in bacterial outbreaks (p < 0.001) CONCLUSION: Norovirus outbreaks were larger but less frequent than bacterial outbreaks, suggesting that underreporting is greater for NoV outbreaks. Food handlers should receive training on the transmission of infections in diverse situations. Very strict control measures on handwashing and environmental disinfection should be adopted in closed or partially-closed institutions. PMID- 18410690 TI - Coordinated prophylactic surgical management for women with hereditary breast ovarian cancer syndrome. AB - BACKGROUND: Women with BRCA1 or BRCA2 mutations have a substantially increased risk of breast and ovarian cancer compared with the general population. Therefore, prophylactic mastectomy (PM) and bilateral salpingo-oophorectomy (BSO) have been proposed as risk-reduction strategies for BRCA1/2 mutation carriers. We aimed to assess the feasibility of coordinated PM and BSO in hereditary breast ovarian cancer syndrome. METHODS: High risk women for breast and ovarian cancer who underwent coordinated PM and BSO were included in this study. Clinical characteristics and surgical and oncologic outcomes were retrospectively reviewed. RESULTS: Twelve patients underwent coordinated PM and BSO. Ten had history of previous breast cancer. Autologous breast reconstruction was performed in ten patients. The mean age at surgery was 43 (range 34-65). Mean operating time was 9.3 hours (range 3-16) with a mean postoperative hospitalization of 5.4 days (range 4-8). Intraoperatively, there were no major surgical complications. Postoperatively, one patient developed an abdominal wound dehiscence, another reoperation for flap congestion; one had umbilical superficial epidermolysis, and one patient developed aspiration pneumonia. At a mean follow-up of 84 months, 10 of patients were cancer-free. Although no patients developed a new primary cancer, two developed a distant recurrence. CONCLUSION: Coordinated PM and BSO is a feasible procedure with acceptable morbidity in selected high-risk patients that desire to undergo surgery at one operative setting. PMID- 18410691 TI - A probe-treatment-reference (PTR) model for the analysis of oligonucleotide expression microarrays. AB - BACKGROUND: Microarray pre-processing usually consists of normalization and summarization. Normalization aims to remove non-biological variations across different arrays. The normalization algorithms generally require the specification of reference and target arrays. The issue of reference selection has not been fully addressed. Summarization aims to estimate the transcript abundance from normalized intensities. In this paper, we consider normalization and summarization jointly by a new strategy of reference selection. RESULTS: We propose a Probe-Treatment-Reference (PTR) model to streamline normalization and summarization by allowing multiple references. We estimate parameters in the model by the Least Absolute Deviations (LAD) approach and implement the computation by median polishing. We show that the LAD estimator is robust in the sense that it has bounded influence in the three-factor PTR model. This model fitting, implicitly, defines an "optimal reference" for each probe-set. We evaluate the effectiveness of the PTR method by two Affymetrix spike-in data sets. Our method reduces the variations of non-differentially expressed genes and thereby increases the detection power of differentially expressed genes. CONCLUSION: Our results indicate that the reference effect is important and should be considered in microarray pre-processing. The proposed PTR method is a general framework to deal with the issue of reference selection and can readily be applied to existing normalization algorithms such as the invariant-set, sub array and quantile method. PMID- 18410692 TI - Self-testing for cancer: a community survey. AB - BACKGROUND: Cancer-related self-tests are currently available to buy in pharmacies or over the internet, including tests for faecal occult blood, PSA and haematuria. Self-tests have potential benefits (e.g. convenience) but there are also potential harms (e.g. delays in seeking treatment). The extent of cancer related self-test use in the UK is not known. This study aimed to determine the prevalence of cancer-related self-test use. METHODS: Adults (n = 5,545) in the West Midlands were sent a questionnaire that collected socio-demographic information and data regarding previous and potential future use of 18 different self-tests. Prevalence rates were directly standardised to the England population. The postcode based Index of Multiple Deprivation 2004 was used as a proxy measure of deprivation. RESULTS: 2,925 (54%) usable questionnaires were returned. 1.2% (95% CI 0.83% to 1.66%) of responders reported having used a cancer related self test kit and a further 36% reported that they would consider using one in the future. Logistic regression analyses suggest that increasing age, deprivation category and employment status were associated with cancer related self-test kit use. CONCLUSION: We conclude that one in 100 of the adult population have used a cancer-related self-test kit and over a third would consider using one in the future. Self-test kit use could alter perceptions of risk, cause psychological morbidity and impact on the demand for healthcare. PMID- 18410693 TI - A general approach to simultaneous model fitting and variable elimination in response models for biological data with many more variables than observations. AB - BACKGROUND: With the advent of high throughput biotechnology data acquisition platforms such as micro arrays, SNP chips and mass spectrometers, data sets with many more variables than observations are now routinely being collected. Finding relationships between response variables of interest and variables in such data sets is an important problem akin to finding needles in a haystack. Whilst methods for a number of response types have been developed a general approach has been lacking. RESULTS: The major contribution of this paper is to present a unified methodology which allows many common (statistical) response models to be fitted to such data sets. The class of models includes virtually any model with a linear predictor in it, for example (but not limited to), multiclass logistic regression (classification), generalised linear models (regression) and survival models. A fast algorithm for finding sparse well fitting models is presented. The ideas are illustrated on real data sets with numbers of variables ranging from thousands to millions. R code implementing the ideas is available for download. CONCLUSION: The method described in this paper enables existing work on response models when there are less variables than observations to be leveraged to the situation when there are many more variables than observations. It is a powerful approach to finding parsimonious models for such datasets. The method is capable of handling problems with millions of variables and a large variety of response types within the one framework. The method compares favourably to existing methods such as support vector machines and random forests, but has the advantage of not requiring separate variable selection steps. It is also works for data types which these methods were not designed to handle. The method usually produces very sparse models which make biological interpretation simpler and more focused. PMID- 18410694 TI - Food prices, inequity, and our responsibilities. PMID- 18410695 TI - Do national research priorities align with burden of disease? PMID- 18410696 TI - Out of the box. Public Health Nutrition. PMID- 18410697 TI - Feasibility of using retinol-binding protein from capillary blood specimens to estimate serum retinol concentrations and the prevalence of vitamin A deficiency in low-resource settings. AB - Vitamin A deficiency (VAD) is a significant public health problem in many countries. While cost-effective interventions are available to control VAD, reliable information is needed to the track progress of control programmes. However, assessment of VAD is uncommon because current approaches are expensive and not feasible in low-resource settings. The present study explores the utility of retinol-binding protein (RBP), analysed by enzyme-linked immunosorbent assay from capillary blood, as an alternative measure of serum retinol concentrations in populations. The study collected matched panels of venous and capillary blood from pre-school children in Chiang Mai, Thailand. Of a total sample of 195 children, there were no differences between RBP from venous blood, RBP from capillary blood or retinol from capillary blood relative to retinol from venous blood. Receiver-operating characteristic curve analysis suggested a cut-off of RBP < 0.825 micromol l(-1) had optimal screening proficiency relative to retinol <0.70 micromol l(-1). For the purpose of population assessment, all three parameters performed well in screening for VAD relative to retinol from venous blood. There were no differences in the estimates of VAD between children stratified by inflammation status. Lower RBP concentrations were found in children in the early convalescent stage of infection than in children with no infection or in the late convalescent stage. This study provided evidence of the biological comparability between retinol and RBP estimated from venous blood and capillary blood. This is a critical observation as it provides empirical evidence that RBP from capillary blood is a surrogate measure of serum retinol concentrations. PMID- 18410698 TI - Functional foods or functionally marketed foods? A critique of, and alternatives to, the category of 'functional foods'. PMID- 18410699 TI - Alcohol and illicit drug dependence among parents: associations with offspring externalizing disorders. AB - BACKGROUND: Previous research indicates that alcohol and drug dependence constitute aspects of a general vulnerability to externalizing disorders that accounts for much of the parent-offspring resemblance for these and related disorders. This study examined how adolescent offspring risk for externalizing psychopathology varies with respect to parental alcoholism and illicit drug dependence. METHOD: Data from the Minnesota Twin Family Study, a community-based investigation of adolescents (age 17 years, n=1252) and their parents, were used. Lifetime diagnoses of alcohol and drug dependence (among both parents and offspring) and offspring attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, adult antisocial behavior, and nicotine dependence were assessed via structured interviews. RESULTS: Parental alcohol dependence and parental drug dependence were similarly associated with increased risk for nearly all offspring disorders, with offspring of alcohol and drug dependent parents having approximately 2-3 times the odds for developing a disorder by late adolescence compared to low-risk offspring. Compared to parental dependence on other illicit drugs, parental cannabis dependence was associated with weaker increased risk for offspring externalizing disorders. CONCLUSIONS: Both parental alcohol and drug dependence are independently associated with an increased risk for a broad range of externalizing psychopathology among late adolescent offspring. PMID- 18410700 TI - The relationship of dementia prevalence in older adults with intellectual disability (ID) to age and severity of ID. AB - BACKGROUND: Previous research has shown that adults with intellectual disability (ID) may be more at risk of developing dementia in old age than expected. However, the effect of age and ID severity on dementia prevalence rates has never been reported. We investigated the predictions that older adults with ID should have high prevalence rates of dementia that differ between ID severity groups and that the age-associated risk should be shifted to a younger age relative to the general population. METHOD: A two-staged epidemiological survey of 281 adults with ID without Down syndrome (DS) aged 60 years; participants who screened positive with a memory task, informant-reported change in function or with the Dementia Questionnaire for Persons with Mental Retardation (DMR) underwent a detailed assessment. Diagnoses were made by psychiatrists according to international criteria. Prevalence rates were compared with UK prevalence and European consensus rates using standardized morbidity ratios (SMRs). RESULTS: Dementia was more common in this population (prevalence of 18.3%, SMR 2.77 in those aged 65 years). Prevalence rates did not differ between mild, moderate and severe ID groups. Age was a strong risk factor and was not influenced by sex or ID severity. As predicted, SMRs were higher for younger age groups compared to older age groups, indicating a relative shift in age-associated risk. CONCLUSIONS: Criteria-defined dementia is 2-3 times more common in the ID population, with a shift in risk to younger age groups compared to the general population. PMID- 18410701 TI - Dual task performance in early Alzheimer's disease, amnestic mild cognitive impairment and depression. AB - BACKGROUND: The dual task paradigm (Baddeley et al. 1986; Della Sala et al. 1995) has been proposed as a sensitive measure of Alzheimer's dementia, early in the disease process. METHOD: We investigated this claim by administering the modified dual task paradigm (utilising a pencil-and-paper version of a tracking task) to 33 patients with amnestic mild cognitive impairment (aMCI) and 10 with very early Alzheimer's disease, as well as 21 healthy elderly subjects and 17 controls with depressive symptoms. All groups were closely matched for age and pre-morbid intellectual ability. RESULTS: There were no group differences in dual task performance, despite poor performance in episodic memory tests of the aMCI and early Alzheimer's disease groups. In contrast, the Alzheimer patients were specifically impaired in the trail-making test B, another commonly used test of divided attention. CONCLUSIONS: The dual task paradigm lacks sensitivity for use in the early differential diagnosis of Alzheimer's disease. PMID- 18410702 TI - Association of BMI and nutritional habits with hypertension in the adult population of Croatia. AB - OBJECTIVE: The purpose of the present study was to investigate the distribution of hypertension, its nutritional and other determinants and their association in Croatia. DESIGN: Population-based, cross-sectional study. SETTING: Croatia. SUBJECTS: Representative random sample of 9070 adults (18 years and older) from all Croatian regions, selected using multistage sampling. RESULTS: Overall prevalence of hypertension (arterial blood pressure >or=140/90 mmHg) in Croatia was 45.5 %, of which 40 % were newly diagnosed hypertensives. There was no difference in the percentage of hypertensives between the continental and Mediterranean parts of Croatia. Newly diagnosed hypertension was found to be positively associated with increased BMI, age and alcohol consumption in the total sample and with moderate consumption of coffee in women, as well as inversely associated with regular consumption of fruits and vegetables in the total sample and high consumption of coffee in the total sample and in men. No association with newly diagnosed hypertension was recorded for use of animal fat, consumption of whole milk, smoked meat products, for adding salt, smoking and leisure-time physical activity. CONCLUSIONS: Hypertension is a major public health problem in both continental and Mediterranean parts of Croatia. Maintenance of normal body mass, its reduction if already increased, and regular consumption of fruits and vegetables are proposed as priorities for primary prevention of hypertension in Croatia. PMID- 18410703 TI - Low dietary calcium in European postmenopausal osteoporotic women. AB - OBJECTIVE: The WHO recommends a daily Ca intake for postmenopausal women of 1300 mg. The objective of the present study was to assess the dietary Ca intake in European postmenopausal osteoporotic women. DESIGN, SETTING AND SUBJECTS: Assessment of dietary Ca intake (food and supplements) was performed with a validated self-questionnaire in 8524 osteoporotic women from nine European countries (Belgium, Denmark, France, Germany, Hungary, Italy, Poland, Spain and the UK). RESULTS: Mean age of the patients was 74.2 (sd 7.1) years, mean BMI was 25.7 (sd 4.2) kg/m2. Of the study population, 37.2 % of the women took Ca supplements. The mean dietary intake of Ca was 930.7 (sd 422.9) mg/d. The lowest Ca intake was found in Hungary (586.7 (sd 319.1) mg/d) and the highest in Denmark (1145.6 (sd 463.0) mg/d). In the whole study population, only 19.1 % of the women had a dietary Ca intake >1300 mg/d. Only 17.1 % of women aged over 75 years achieved 1300 mg/d compared with 20.5 % of women aged less than 75 years (P = 0.0001 for the difference between the two groups). CONCLUSION: Dietary intake of Ca is very low in European postmenopausal women. A greater awareness is needed to resolve this public health problem. PMID- 18410704 TI - Intakes of fruits, vegetables and carbohydrate and the risk of CVD. AB - BACKGROUND: Low-carbohydrate diets could lead to reduced fruit and vegetable intake, which may be protective against CVD. The role of carbohydrate intake in modifying the association between fruits and vegetables and CVD has not been evaluated. OBJECTIVE: To evaluate whether carbohydrate intake affects the association between fruits and vegetables and CVD. DESIGN: We included participants from two large prospective studies, the Nurses' Health Study (NHS) and the Health Professionals' Follow-Up Study (HPFS). We followed 70 870 eligible NHS females for 16 years and 38 918 eligible HPFS males for 14 years. Diet was assessed from an FFQ updated every 4 years. Our primary outcome was ischaemic CVD (fatal and non-fatal myocardial infarction and ischaemic stroke). We used Cox proportional hazards models to evaluate the relationship between fruits and vegetables and ischaemic CVD within groups with low, moderate or high carbohydrate intake. RESULTS: Fruit intake was strongly related with carbohydrate intake, but vegetables showed a very small correlation. Vegetable intake showed stronger associations with ischaemic CVD among the low carbohydrate group (multivariate risk ratio (RR) = 0.82 for an increment of 3 servings/d; 95 % CI 0.68, 0.99); green leafy vegetables and carotene-rich fruits and vegetables followed a similar pattern. Total fruit intake was associated with a lower risk of ischaemic CVD only among participants with moderate carbohydrate intake (RR = 0.81 comparing extreme quintiles; 95 % CI 0.70, 0.94). CONCLUSIONS: Total vegetables, green leafy vegetables and carotene-rich fruits and vegetables showed stronger associations with ischaemic CVD among the low carbohydrate group. No consistent trends were observed for fruit intake. PMID- 18410705 TI - Impact of chewing betel-nut (Areca catechu) on liver cirrhosis and hepatocellular carcinoma: a population-based study from an area with a high prevalence of hepatitis B and C infections. AB - BACKGROUND: Chewing betel-nuts (Areca catechu) is carcinogenic but the risk for hepatocellular carcinoma (HCC) and liver cirrhosis (LC) is little considered. Worldwide 600 million people chew betel, including emigrants from palm-growing countries. OBJECTIVE: We aimed to assess the relationships and dose-response effects of betel chewing on LC and HCC risks, since habit cessation could reduce the increased risks of HCC and LC found in such communities. SUBJECTS: Screening 60 326 subjects aged 30-79 years in a population-based study in Taiwan identified LC in 588 and HCC in 131 subjects. Demographic features, hepatitis B/C infections, other risk factors and betel chewing were noted. Multiple Cox regression models were used to assess independent relationships, interactions and synergisms between age, betel chewing and hepatitis B/C. RESULTS: Betel chewing increased LC and HCC risk 4.25-fold (95 % CI 2.9, 6.2) in current chewers and 1.89-fold (95 % CI 1.13, 3.16) in ex-chewers v. never-chewers, with dose effects for quantity, duration and cumulative exposure in chewers. Subjects without hepatitis B/C infections had 5.0-fold (95 % CI 2.87, 9.03) increased risk of LC/HCC v. never-chewers, and betel chewing had an additive synergistic effect on hepatitis B/C-related risks. Risk reduction with betel habit cessation could exceed that expected from immunization programmes for hepatitis B and C. CONCLUSION: Increased risks of cirrhosis and hepatocellular cancer were found in betel chewers free of hepatitis B/C infection, and these risks were synergistically additive to those of hepatitis B/C infections. Estimated risk reduction from effective anti-betel chewing programmes would be sizeable. PMID- 18410706 TI - Prevalence of obesity and overweight among urban adults in Bulgaria. AB - OBJECTIVE: The prevalence of obesity is increasing in transitional countries, but extensive data on some countries, such as Bulgaria, are still lacking. Therefore, the objective of the present study was to estimate the prevalence of overweight and obesity, relative to gender and income, among adults in Sofia, Bulgaria. DESIGN: Cross-sectional survey to collect data on diet, health, BMI and income using a brief questionnaire on diet and income. Data were analysed using analysis of covariance to determine differences within and between income and gender groups. SETTING: Sofia, Bulgaria. SUBJECTS: Adults living in the city of Sofia, Bulgaria. RESULTS: For adults 30-60 years of age, 35.1 % were overweight and 6.2 % were obese. The proportion of overweight and obesity was higher among men than women (44.8 % v. 32.4 % and 6.0 % v. 4.7 %, respectively). With respect to income, BMI decreased as income increased. For men, BMI was highest for the lowest and highest income groups, whereas for women lower income was associated with a higher BMI. CONCLUSION: The prevalence of overweight and obesity appears to be an emerging problem in some sectors of Bulgarian society, based on our data from the largest urban area of the country. These data provide new information on the divergence in health and disease risk in a country that is still economically challenged and may be facing the nutrition transition. PMID- 18410707 TI - Defining 'low in fat' and 'high in fat' when applied to a food. AB - OBJECTIVES: To describe four different methods of identifying indicator foods that are high, medium or low in fat with reference to dietary patterns and to use these indicator foods to test three sets of definitions of 'high', 'medium' and 'low' in fat from 'banding schemes' developed by the Coronary Prevention Group (CPG), the Food Standards Agency (FSA) and Sainsbury's. METHODS: Indicator foods were developed using food intake data from the UK National Diet and Nutrition Survey and two parameters: (i) probability of the food being consumed by an individual with a high-fat diet (Method 1); and (ii) the contribution of the food to the fat intake of the average diet of consumers (Methods 3 and 4). Method 2 used both parameters. The three banding schemes were tested by assessing their levels of agreement with methods in categorising indicators. RESULTS: Sensitivity in identifying high, medium and low fat indicators was highest with the CPG banding scheme (high and medium fat indicators) and Sainsbury's scheme (low fat indicators) (Methods 2, 3 and 4). The levels of agreement (kappa coefficient) were 0.68 for the CPG scheme; 0.51 for the Sainsbury's scheme; and 0.41 for the FSA scheme (Method 3). CONCLUSIONS: It is possible to use indicator foods related to dietary patterns of a specific population to generate more rational definitions of 'high', 'medium' and 'low' in fat. This could be the starting point for the development of indicator foods for testing more complex nutrient profile models (i.e. those that consider more than one nutrient). PMID- 18410708 TI - Urinary tract infection and adverse outcome of pregnancy. AB - Urinary tract infections (UTIs) during pregnancy are among the commonest health problems world wide, specially in developing countries, including Egypt. It has several adverse outcomes not only on the mother but also on the fetus as well.. The aim of this study is to determine the incidence of UTIs during pregnancy, study the main risk factors associated with such infections and find the impact of these infections on some pregnancy outcomes namely the gestational age and birth weight. A follow-up study on 249 pregnant women attending the ante natal care clinic at Zagazig university hospital. They were recruited over a period from 1st of September to 30th of or November, 2005. The outcome could be recorded for 201 of them. Data were collected through a pretested questionnaire, repeated urine analyses and recording of outcome of pregnancy. The study revealed that the incidence of UTIs during pregnancy was 31.3%. The commonest organisms were Klebsiella and E-coli. Several socio-demographic characteristics were found significantly associated with UTIs, age being 30 years and more, illiterates and low educational level, low socio-economic level and those with unsatisfactory personal hygiene and those using underwear clothes other than cotton. Significant associations with UTIs were also found in multigravidae 4th and more, those having more than one child and those who previously suffered UTIs. The only predicting variable with UTIs was low socio-economic level. The study revealed that the probability of delivering premature infants and low birth weights was significantly higher among those who experienced UTIs during pregnancy. Multivariate analysis revealed that UTI was one of the main contributors to pre mature deliveries. CONCLUSION AND RECOMMENDATIONS: Urinary tract infections with pregnancy still constitute a big problem with high incidence. It has a great impact on pregnancy outcome mainly pre-mature labor. So, the study recommends health education about personal hygiene, repeated urine cultures early in pregnancy and during the third trimesters, specially for low socio-economic pregnant women. Specific antibiotics should be prescribed to avoid adverse pregnancy outcome. PMID- 18410709 TI - Assessment of health related quality of life among school children with cancer in Alexandria. AB - In recent years, the necessity of measuring health related quality of life (HRQOL) among children with cancer has been stressed. The aim of the present work was to assess the HRQOL in terms of physical, emotional and social functioning, to identify some predictors of QOL and to describe the scholastic achievement and nutritional status among school children with cancer in Alexandria. A cross sectional study of 215 students with cancer who attended the oncology school health insurance clinic of Sporting Student's Hospital in Alexandria governorate within a period of one month (June 2005) was conducted. Every child was subjected to the Pediatric Oncology Quality Of Life scale to assess the quality of life with cancer. Anthropometric measurements including weight and height were measured for each child and BMI was calculated. Review of records of each child was done to take the last heamoglobin level. Lymphoma was the most common tumour (34.9%), followed by leukemia (24.2%), brain tumour (13.0%), osteosarcoma (7.9%), and Wilm's tumour (7%). About one third of the sample (36.7%) reported poor QOL which was more evident in the physical domain (54.4%), as compared to 34.4% in emotional domain and 26% in the social domain. Sex (p<0.05), age (p<0.05), complications (p<0.01), place of treatment (p<0.05), place of diagnosis (p<0.05) and treatment (p<0.05) were all significantly associated with QOL. However after adjustment for other variables, poor QOL was more likely among students of younger age, students with complications, and students whose diagnosis was done at the student hospital. About two thirds of the sample (68.5 %) reported poor scholastic achievement, 25.1% were underweight and 87.4% were anemic. CONCLUSION AND RECOMMENDATIONS: HRQOL measurement should be done as a routine for all children with cancer in order to identify specific therapies that require modifications in management. More efforts are needed to improve the health care in the student's hospital, especially for young children with cancer. PMID- 18410710 TI - Nutritional factors affecting the menarcheal state of adolescent school girls in Alexandria. AB - The nutritional status of adolescents plays a dominant role in determining the maturation status. The aim of the present work was to assess the mean age of menarche and the main nutritional factors affecting it. A cross-sectional study of 1606 girls was conducted in primary and preparatory schools in Alexandria. Every girl was subjected to anthropometric assessment including weight, height, mid upper arm circumference (MUAC), waist circumference, hip circumference and triceps skin-fold thickness. BMI and body fat percentage were calculated. A 24 hours diet recall method was used to assess the dietary intake. The mean age of menarche was 11.98+/-0.96 years. The mean MUAC, triceps skin-fold thickness, waist circumference and hip circumference were significantly higher among menstruating girls as compared to non-menstruating. (p<0.01). Only 7.5% of the females less than the 5th percentile of BMI (thinness) were menstruating, while the corresponding figure for those at or more than 85th percentile (overweight) was 65.6% and this was statistically significant (X 2 (2) =102.8, P<0.001). Girls who attained menstruation demonstrated a higher significant mean percent of body fat (43.40+/-10.0) as compared to non menstruating ones (35.41+/-7.87), ( t=17.09, P<0.001). The oldest age at menarche was noted when the protein, iron and caloric intake was less than 80% of the RDAs. However after adjustment of other variables direct relation was detected between age of girls and their age of menarche and those in private school had earlier age of menarche than those in governmental one. The nutritional status of the adolescents had a significant association with the onset of menstruation and the age at menarche. PMID- 18410711 TI - Anemia among female college students attending the University of Sharjah, UAE: prevalence and classification. AB - Anemia remains a public health challenge worldwide. Very few studies have been conducted on anemia in the United Arab Emirates and they have focused on children and pregnant women. Little is known about anemia among college female students from a preliminary study of iron deficiency anemia that was conducted on medical college students. This study aimed to estimate the prevalence of anemia among female college students attending the University of Sharjah (UoS) in the United Arab Emirates (UAE). A secondary analysis of the records of 258 complete blood cell count results from consented female college students. Hemoglobin (Hb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) were used to determine the prevalence of and classify anemia according to RBC indices and disease severity. Emirati students represented 50.8% of the studied population. The overall prevalence of anemia (Hb <12g/dL) was 26.7% and the majority (88.4%) of the 69 anemic students had mild anemia, whereas 7.2% were moderately anemic and 2.3% Emirati students were severely anemic (hemoglobin <7g/dL). About 15.9% of the anemia cases were microcytic (MCV <80fL) and 1.6% were macrocytic (MCV >96fL). The results showed that anemia constitutes a health problem among female college students in the UAE, and most of the detected anemia was microcytic with mild severity. This warrants further study on a larger sample of healthy college students to validate these findings and eventually encourage the development of directed educational and nutritional programs to safeguard the health of these future mothers. PMID- 18410712 TI - Impact of helicobacter pylori infection on the activities of urease and lipase enzymes in patients with giardiasis. AB - The study of the effect of Giardia lamblia and Helicobacter pylori organisms coexistence on the activities of urease and lipase enzymes was the aim of this work which was done through choosing 50 cases of giardiasis in addition to 10 normal individuals chosen as a control group (free from giardiasis). It is well known that H. pylori is considered one of the most important causes of gastric and duodenal inflammations which could predispose to ulcers and hypochlorhydria leading to increased susceptibility to giardiasis as it is known that HCl acts as a chemical barrier to microbes. The biochemical tests were done to investigate the activity of both urease and lipase enzymes extracted from the gastric juice of patients and controls. A significant increase in urease activity in the group having combined infection (giardiasis and H.pylori) than the group infected with G.lamblia alone and the control group was found. The same findings were obtained regarding the lipase activity. In the present work, both infections H. pylori and G. lamblia coexisted in 75% of epigastric pain cases which could be explained on the basis that both organisms predispose to each other. PMID- 18410713 TI - Seroprevalence of measles, mumps, rubella and varicella among staff of a hospital in Riyadh, Saudi Arabia. AB - This study aimed at screening immune status of Health Care Workers (HCWs) against measles, mumps, rubella, and varicella, and assessing the reliability of the history of previous illness due to these infections or vaccination against them as an indicator of the immune status, as checked by laboratory tests. Demographic data, history of previous illnesses and previous vaccinations were collected using a self-administered questionnaire. Also, serologic screening was done for these infections. Antibodies tested using ELISA. Among tested hospital staff, 71.8%, 60.3%, 47.9% and 68.4% reported history of infection or vaccination against measles, mumps, rubella and varicella respectively, while laboratory results proved that 4.5%, 10.8%, 12.9% and 11.3% were susceptible respectively. Susceptibility was most frequent among housekeeping staff against measles and varicella, while administrative staff were the most susceptibles to mumps, and nurses were the most susceptibles to rubella. Sensitivity of past history of illness as an indicator of the immune status, was found to be (85%, 83%, 66%, 86%); Specificity was (81%, 81%, 89%, 87%); Positive Predictive value (PV) was (99%, 98%, 98%, 98%) and Negative PV was (16%, 29%, 22%, 39%) for measles, mumps, rubella and varicella respectively. Screening for measles, mumps, rubella and varicella among hospital staff is mandatory to detect those who are susceptible for infections and should be vaccinated, and so preventing transmission of these infections to their colleagues or patients. PMID- 18410714 TI - Cesarean section deliveries in one health insurance hospital in Alexandria. AB - Cesarean section (CS) rates have been increasing world wide, raising the question of the appropriateness of the selection of cases for the procedure. The World Health Organization (WHO) states that no region in the world is justified in having a cesarean rate greater than 10 to 15 percent. The aim of the work was to determine the trend of cesarean section deliveries in Gamal Abdel Naser Hospital which is affiliated to the Health Insurance Organization (HIO). The study was conducted through a descriptive retrospective approach. The study sample included the a)recorded deliveries between 1998-2005 (n=15917) for estimating the trend of cesarean section deliveries, and b) the medical records of CS deliveries at 2002 in the hospital (n=837) for identifying the indications of CS and their adequacy as a source of information for evaluation of CS deliveries . The study revealed that; cesarean section rate was high and increasing during the period from 1998 - 2005. The highest percent was in the year 2004 (57.9%). The trend of increase was significant (c for linear trend = 162.717, p= 0.000). Thursdays accounted for the highest percent of both admissions and deliveries, while Fridays accounted for the lowest percent . More than one half of deliveries occurred between 2 pm to before 8 pm. More than three quarters of the study sample (77.9%) did not have trial labour. Only 12.8% of the total study sample had induction and the outcome of induction was dystocia in 85%. The main indication of cesarean section was previous CS (41.2%), fetal distress (17.6%), failed trial and failure to progress (11.4%), cephalo-pelvic disproportion (10.3%), abnormal presentation (5.6%) and ante-partum hemorrhage (3.2%). Patient's records lack most of the essential information so it was not possible to verify recorded indication to justify caesarean section. PMID- 18410715 TI - Health impacts on workers in landfill in Jeddah City, Saudi Arabia. AB - All workers (twenty nine workers) in the Jeddah landfill were interviewed. They were asked to answer a precoded questionnaire. Jeddah landfill lies in Breman region on the east of the highway. It was constructed on the year 1982. Its total area is about 1.5 square kilometers. The daily solid waste weight disposed off in the landfill is from three to four thousands tons. There is a network of pipes and wells to collect gases which are burnt. There is no network for the disposal of the leachate. The personnel are 29 they work in shifts. The majority (62.1%) are from Bangladesh. This can be explained by the fact that they are poor, ignorant and work in any job regardless the hazards that my result from such work. It was found that 17.9% had accidents. Errors in dealing with instruments and falls each constitutes 20% of accidents' causes. Other causes represent 60%. The major four health problems are respiratory infections and/or allergy, eye infections, gastrointestinal tract (GIT) infections, and musculoskeletal injuries representing 65.5%, 48.3%, 20.7%, and 17.2%, respectively. It is recommended for landfill workers to wear protective clothes and equipment, have work-related training courses, and be medically supervised. PMID- 18410716 TI - Assessment of the mean daily dietary intake of acrylamide in alexandria. AB - Acrylamide is a toxic and potentially cancer causing chemical formed in thermally treated starchy foods. The objective of the present study is to estimate the mean daily dietary intake of acrylamide in Alexandria governorate. A total of 822 subjects (417 males and 405 females) were randomly selected, from different places such as clubs, schools, university, nursery schools, homes and companies, from different socio-demographic sectors of the Alexandria population. The age of the subjects ranged from less than four to more than fifty years old. Subjects were divided into seven age groups. A twenty four hours recall was used to record types and quantities of the food stuffs taken through three consecutive days including the week end. Samples of consumed starchy foods - presumed to contain high levels of acrylamide were collected and analyzed for acrylamide. Thirteen food items were analyzed for acrylamide. The highest acrylamide contents were recorded in chips potato and Fried potatoes (French fries) (1500 +/- 645 and 540 +/- 213 microg/kg respectively). Cakes were the lowest in mean acrylamide contents (12 +/- 12 microg/kg). The highest mean daily dietary intake of acrylamide (3.82 microg/kg body weight) was for the age group from 3 - < 6 years old subjects, while the lowest acrylamide intake (0.49 microg/kg body weight) was that of the age group above 50 years old. The overall estimated mean daily dietary intake of acrylamide was 1.75 microg/kg body weight. No significant differences between males and females were found among various studied age groups regarding the mean daily dietary intake of acrylamide. Chips potato represents the major source of dietary acrylamide (46%). The second most contributing agent in the mean daily dietary intake of acrylamide for studied Alexandria population was fried potato (19%). Bread contributes to 17 % of the mean daily dietary acrylamide intake. In conclusion, the fried and deep fried starchy foods consumed by the studied Alexandria population contain high levels of acrylamide. The majority of the estimated mean dietary daily intake of acrylamide is from fried potato products. Controlling the intake, especially in young children can thus reduce the high acrylamide intake. PMID- 18410717 TI - Factors associated with geographic migration among a cohort of injection drug users. AB - We sought to determine factors associated with migration among injection drug users in Vancouver, Canada. We examined migration patterns among participants in the Vancouver Injection Drug Users Study. All participants were residents of Vancouver at the time of recruitment. Correlates of migration, defined as living outside of Greater Vancouver between June 1999 and May 2005, were identified using generalized estimating equations. Various factors were negatively associated with migration including frequent crack cocaine smoking (AOR = 0.44, 95% CI: 0.37-0.52), current methadone use (AOR = 0.50, 95% CI: 0.40-0.63), frequent heroin injection (AOR = 0.51, 95% CI: 0.41-0.64), requiring help injecting (AOR = 0.60, 95% CI: 0.47-0.77), sex trade involvement (AOR = 0.64, 95% CI: 0.51-0.82), living in unstable housing (AOR = 0.69, 95% CI: 0.58-0.83), public injecting (AOR = 0.75, 95% CI: 0.60-0.94), and incarceration (AOR = 0.77, 95% CI: 0.61-0.96). Alcohol use was positively associated with migration in this analysis (AOR = 1.25, 95% CI: 1.05-1.48). Our findings suggest that participants who migrated were less at risk for HIV infection, given lower levels of reported risk-taking. PMID- 18410719 TI - Resolving animal phylogeny: a sledgehammer for a tough nut? AB - While great progress toward discovering the evolutionary relationships of major animal groups has been made in the past 2 decades, significant portions of the animal tree remain unresolved. A recent manuscript in Nature by Dunn and colleagues tackles this problem using massive sequence data sets from many taxa the so-called "phylogenomic" approach. PMID- 18410718 TI - Patterning lessons from a dorsalized embryo. AB - A paper by Nunes da Fonseca and colleagues in this issue of Developmental Cell shows that, to pattern its dorsoventral axis, the beetle Tribolium utilizes many of the same genes used in flies, but in very different ways: rather than relying on maternal information, it uses Dorsal and Dpp as part of two coordinated ancestral self-organized systems. PMID- 18410720 TI - When ATPases pontin and reptin met telomerase. AB - Pontin and reptin are conserved AAA+ ATPases identified in chromatin-remodeling complexes. In a recent issue of Cell, Venteicher et al. provide new insight into the function of pontin and reptin in telomerase biogenesis, which is important for cellular senescence, aging, and cancer. These unexpected findings have implications for new avenues for development of effective therapeutic drugs in human disease. PMID- 18410721 TI - Brushed aside and silenced. AB - Mammalian genomes are highly organized in the 3D space of cell nuclei, but whether this affects gene function is unclear. Three papers now show that spatial relocation of a gene directly affects expression, and surprisingly, that of its neighbors. PMID- 18410722 TI - Lectins in the endoplasmic reticulum: the sweet side of protein quality control. AB - In a recent study in Nature Cell Biology, Christianson et al. provide intriguing insights into the mechanisms of mammalian protein quality control in the endoplasmic reticulum. Their findings open up new perspectives on the versatility and diversity of how protein quality control sorts out defective polypeptides to prevent damage to the cell. PMID- 18410723 TI - To thine own self be true: self-fusion in single-celled tubes. AB - In this issue of Developmental Cell, Rasmussen et al. (2008) investigate the morphogenesis of the Caenorhabditis elegans pharynx. Their results highlight the usefulness of this system for investigating the molecular mechanisms behind the unusual cell behaviors that underlie the formation of single-celled tubes during animal development. PMID- 18410724 TI - Plant hormones and signaling: common themes and new developments. AB - About 200 plant biologists convened in Keystone, Colorado, for the "Plant Hormones and Signaling" symposium, which was organized by Joanne Chory, Joe Ecker, and Mark Estelle. The meeting was run concurrently with the "Plant Innate Immunity" symposium organized by Jonathan Jones and Jane Glazebrook. In this report, we summarize the progress in plant hormones and signaling. PMID- 18410725 TI - Linking kinetochore-microtubule binding to the spindle checkpoint. AB - The spindle checkpoint blocks cell-cycle progression until chromosomes are properly attached to the mitotic spindle. Popular models propose that checkpoint proteins associate with kinetochores to produce a "wait anaphase" signal that inhibits anaphase. Recent data suggest that a two-state switch results from using the same kinetochore proteins to bind microtubules and checkpoint proteins. At least eight protein kinases are implicated in spindle checkpoint signaling, arguing that a traditional signal transduction cascade is integral to spindle checkpoint signaling. PMID- 18410726 TI - Epigenetic blocking of an enhancer region controls irradiation-induced proapoptotic gene expression in Drosophila embryos. AB - Drosophila embryos are highly sensitive to gamma-ray-induced apoptosis at early but not later, more differentiated stages during development. Two proapoptotic genes, reaper and hid, are upregulated rapidly following irradiation. However, in post-stage-12 embryos, in which most cells have begun differentiation, neither proapoptotic gene can be induced by high doses of irradiation. Our study indicates that the sensitive-to-resistant transition is due to epigenetic blocking of the irradiation-responsive enhancer region (IRER), which is located upstream of reaper but is also required for the induction of hid in response to irradiation. This IRER, but not the transcribed regions of reaper/hid, becomes enriched for trimethylated H3K27/H3K9 and forms a heterochromatin-like structure during the sensitive-to-resistant transition. The functions of histone-modifying enzymes Hdac1(rpd3) and Su(var)3-9 and PcG proteins Su(z)12 and Polycomb are required for this process. Thus, direct epigenetic regulation of two proapoptotic genes controls cellular sensitivity to cytotoxic stimuli. PMID- 18410727 TI - Otefin, a nuclear membrane protein, determines the fate of germline stem cells in Drosophila via interaction with Smad complexes. AB - Nuclear envelope proteins play important roles in chromatin organization, gene regulation, and signal transduction; however, the physiological role of these proteins remains elusive. We found that otefin (ote), which encodes a nuclear lamin-binding protein [corrected], is essential for germline stem cell (GSC) maintenance. We show that Ote, as an intrinsic factor, is both necessary and sufficient to regulate GSC fate. Furthermore, we demonstrate that ote is required for the Dpp/BMP signaling pathway to silence bam transcription. By structure function analysis, we demonstrate that the nuclear membrane localization of Ote is essential for its role in GSC maintenance. Finally, we show that Ote physically interacts with Medea/Smad4 at the bam silencer element to regulate GSC fate. Thus, we demonstrate that specific nuclear membrane components mediate signal-dependent transcriptional effects to control stem cell behavior. PMID- 18410728 TI - Inactivation of a human kinetochore by specific targeting of chromatin modifiers. AB - We have used a human artificial chromosome (HAC) to manipulate the epigenetic state of chromatin within an active kinetochore. The HAC has a dimeric alpha satellite repeat containing one natural monomer with a CENP-B binding site, and one completely artificial synthetic monomer with the CENP-B box replaced by a tetracycline operator (tetO). This HAC exhibits normal kinetochore protein composition and mitotic stability. Targeting of several tet-repressor (tetR) fusions into the centromere had no effect on kinetochore function. However, altering the chromatin state to a more open configuration with the tTA transcriptional activator or to a more closed state with the tTS transcription silencer caused missegregation and loss of the HAC. tTS binding caused the loss of CENP-A, CENP-B, CENP-C, and H3K4me2 from the centromere accompanied by an accumulation of histone H3K9me3. Our results reveal that a dynamic balance between centromeric chromatin and heterochromatin is essential for vertebrate kinetochore activity. PMID- 18410729 TI - Golgi localization of glycosyltransferases requires a Vps74p oligomer. AB - The mechanism of glycosyltransferase localization to the Golgi apparatus is a long-standing question in secretory cell biology. All Golgi glycosyltransferases are type II membrane proteins with small cytosolic domains that contribute to Golgi localization. To date, no protein has been identified that recognizes the cytosolic domains of Golgi enzymes and contributes to their localization. Here, we report that yeast Vps74p directly binds to the cytosolic domains of cis and medial Golgi mannosyltransferases and that loss of this interaction correlates with loss of Golgi localization of these enzymes. We have solved the X-ray crystal structure of Vps74p and find that it forms a tetramer, which we also observe in solution. Deletion of a critical structural motif disrupts tetramer formation and results in loss of Vps74p localization and function. Vps74p is highly homologous to the human GMx33 Golgi matrix proteins, suggesting a conserved function for these proteins in the Golgi enzyme localization machinery. PMID- 18410730 TI - Cell autonomy of HIF effects in Drosophila: tracheal cells sense hypoxia and induce terminal branch sprouting. AB - Drosophila tracheal terminal branches are plastic and have the capacity to sprout out projections toward oxygen-starved areas, in a process analogous to mammalian angiogenesis. This response involves the upregulation of FGF/Branchless in hypoxic tissues, which binds its receptor Breathless on tracheal cells. Here, we show that extra sprouting depends on the Hypoxia-Inducible Factor (HIF)-alpha homolog Sima and on the HIF-prolyl hydroxylase Fatiga that operates as an oxygen sensor. In mild hypoxia, Sima accumulates in tracheal cells, where it induces breathless, and this induction is sufficient to provoke tracheal extra sprouting. In nontracheal cells, Sima contributes to branchless induction, whereas overexpression of Sima fails to attract terminal branch outgrowth, suggesting that HIF-independent components are also required for full induction of the ligand. We propose that the autonomous response to hypoxia that occurs in tracheal cells enhances tracheal sensitivity to increasing Branchless levels, and that this mechanism is a cardinal step in hypoxia-dependent tracheal sprouting. PMID- 18410731 TI - Notch signaling and morphogenesis of single-cell tubes in the C. elegans digestive tract. AB - During organogenesis of the C. elegans digestive system, epithelial cells within a cyst-like primordium develop diverse shapes through largely unknown mechanisms. We here analyze two adjacent, dorsal epithelial cells, called pm8 and vpi1, that remodel their shapes and apical junctions to become donut-shaped, or toroidal, single-cell tubes. pm8 and vpi1 delaminate from the dorsal cyst epithelium and migrate ventrally, across the midline of the cyst, on a transient tract of laminin. pm8 appears to encircle the midline by wrapping around finger-like projections from neighboring cells. Finally, pm8 and vpi1 self-fuse to become toroids by expressing AFF-1 and EFF-1, two fusogens that are each sufficient to promote crossfusion between other cell types. Notch signaling in pm8 induces AFF 1 expression, while simultaneously repressing EFF-1 expression; vpi1 expresses EFF-1 independent of Notch. Thus, the adjacent pm8 and vpi1 cells express different fusogens, allowing them to self-fuse into separate, single-cell tubes while avoiding crossfusion. PMID- 18410732 TI - Collective epithelial migration and cell rearrangements drive mammary branching morphogenesis. AB - Epithelial organs are built through the movement of groups of interconnected cells. We observed cells in elongating mammary ducts reorganize into a multilayered epithelium, migrate collectively, and rearrange dynamically, all without forming leading cellular extensions. Duct initiation required proliferation, Rac, and myosin light-chain kinase, whereas repolarization to a bilayer depended on Rho kinase. We observed that branching morphogenesis results from the active motility of both luminal and myoepithelial cells. Luminal epithelial cells advanced collectively, whereas myoepithelial cells appeared to restrain elongating ducts. Significantly, we observed that normal epithelium and neoplastic hyperplasias are organized similarly, suggesting common mechanisms of epithelial growth. PMID- 18410734 TI - Multiple roles of Notch signaling in the regulation of epidermal development. AB - Recent studies have shown that Notch signaling plays an important role in epidermal development, but the underlying molecular mechanisms remain unclear. Here, by integrating loss- and gain-of-function studies of Notch receptors and Hes1, we describe molecular information about the role of Notch signaling in epidermal development. We show that Notch signaling determines spinous cell fate and induces terminal differentiation by a mechanism independent of Hes1, but Hes1 is required for maintenance of the immature state of spinous cells. Notch signaling induces Ascl2 expression to promote terminal differentiation, while simultaneously repressing Ascl2 through Hes1 to inhibit premature terminal differentiation. Despite the critical role of Hes1 in epidermal development, Hes1 null epidermis transplanted to adult mice showed no obvious defects, suggesting that this role of Hes1 may be restricted to developmental stages. Overall, we conclude that Notch signaling orchestrates the balance between differentiation and immature programs in suprabasal cells during epidermal development. PMID- 18410733 TI - Intra-endodermal interactions are required for pancreatic beta cell induction. AB - The cellular origin of signals that regulate pancreatic beta cell induction is not clearly defined. Here, we investigate the seeming paradox that Hedgehog/Smoothened signaling functions during gastrulation to promote pancreatic beta cell development in zebrafish, yet has an inhibitory role during later stages of pancreas development in amniotes. Our cell transplantation experiments reveal that in zebrafish, Smoothened function is not required in beta cell precursors. At early somitogenesis stages, when the zebrafish endoderm first forms a sheet, pancreatic beta cell precursors lie closest to the midline; however, the requirement for Smoothened lies in their lateral neighbors, which ultimately give rise to the exocrine pancreas and intestine. Thus, pancreatic beta cell induction requires Smoothened function cell-nonautonomously during gastrulation, to allow subsequent intra-endodermal interactions. These results clarify the function of Hedgehog signaling in pancreas development, identify an unexpected cellular source of factors that regulate beta cell specification, and uncover complex patterning and signaling interactions within the endoderm. PMID- 18410735 TI - Self-regulatory circuits in dorsoventral axis formation of the short-germ beetle Tribolium castaneum. AB - The rel/NF-kappaB transcription factor Dorsal controls dorsoventral (DV) axis formation in Drosophila. A stable nuclear gradient of Dorsal directly regulates approximately 50 target genes. In Tribolium castaneum (Tc), a beetle with an ancestral type of embryogenesis, the Dorsal nuclear gradient is not stable, but rapidly shrinks and disappears. We find that negative feedback accounts for this dynamic behavior: Tc-Dorsal and one of its target genes activate transcription of the IkB homolog Tc-cactus, terminating Dorsal function. Despite its transient role, Tc-Dorsal is strictly required to initiate DV polarity, as in Drosophila. However, unlike in Drosophila, embryos lacking Tc-Dorsal display a periodic pattern of DV cell fates along the AP axis, indicating that a self-organizing ectodermal patterning system operates independently of mesoderm or maternal DV polarity cues. Our results also elucidate how extraembryonic tissues are organized in short-germ embryos, and how patterning information is transmitted from the early embryo to the growth zone. PMID- 18410737 TI - Uncoupling Sonic hedgehog control of pattern and expansion of the developing limb bud. AB - Sonic hedgehog (Shh), which regulates proliferation in many contexts, functions as a limb morphogen to specify a distinct pattern of digits. How Shh's effects on cell number relate to its role in specifying digit identity is unclear. Deleting the mouse Shh gene at different times using a conditional Cre line, we find that Shh functions to control limb development in two phases: a very transient, early patterning phase regulating digit identity, and an extended growth-promoting phase during which the digit precursor mesenchyme expands and becomes recruited into condensing digit primordia. Our analysis reveals an unexpected alternating anterior-posterior sequence of normal mammalian digit formation. The progressive loss of digits upon successively earlier Shh removal mirrors this alternating sequence and highlights Shh's role in cell expansion to produce the normal digit complement. PMID- 18410736 TI - Vertebrate CASTOR is required for differentiation of cardiac precursor cells at the ventral midline. AB - The CASTOR (CST) transcription factor was initially identified for its role in maintaining stem cell competence in the Drosophila dorsal midline. Here we report that Xenopus CST affects cardiogenesis. In CST-depleted embryos, cardiomyocytes at the ventral midline arrest and are maintained as cardiac progenitors, while cells in more dorsal regions of the heart undergo their normal program of differentiation. Cardia bifida results from failed midline differentiation, even though cardiac cell migration and initial cell fate specification occur normally. Our fate mapping studies reveal that this ventral midline population of cardiomyocytes ultimately gives rise to the outer curvature of the heart; however, CST-depleted midline cells overproliferate and remain a coherent population of nonintegrated cells positioned on the outer wall of the ventricle. These midline-specific requirements for CST suggest the regulation of cardiomyocyte differentiation is regionalized along a dorsal-ventral axis and that this patterning occurs prior to heart tube formation. PMID- 18410738 TI - Amino acid sensing using aminoacyl-tRNA synthetase. AB - The detection of amino acids using aminoacyl-tRNA synthetases (ARSs) as the molecular recognition element was proposed, and the binding activity and specificity of ARSs were evaluated. Using this rapid and easy method, from 15 to 50 microM tyrosine could be measured specifically. The method suggested in this article could be realized without an amino acid labeling process or a large volume of organic solvents, and the time for measurement was reasonable. PMID- 18410739 TI - HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues. AB - We report robust HPLC/UV methods for quantifying retinyl esters (RE), retinol (ROL), and retinal (RAL) applicable to diverse biological samples with lower limits of detection of 0.7, 0.2, and 0.2 pmol, respectively, and linear ranges greater than 3 orders of magnitude. These assays function well with small, complex biological samples (10-20mg tissue). Coefficients of variation range from 5.9 to 10.0% (intraday) and from 5.9 to 11.0% (interday). Quantification of endogenous RE, ROL, and RAL in mouse serum and tissues (liver, kidney, adipose, muscle, spleen, testis, skin, brain, and brain regions) reveals utility. Ability to discriminate spatial concentrations of ROL and RE is illustrated with C57BL/6 mouse brain loci (hippocampus, cortex, olfactory bulb, thalamus, cerebellum, and striatum). We also developed a method to distinguish isomeric forms of ROL to investigate precursors of retinoic acid. The ROL isomer assay has limits of detection between 3.5 and 4.5 pmol and has a linear range and coefficient of variation similar to those of the ROL/RE and RAL assays. The assays described here provide for sensitive and rigorous quantification of endogenous RE, ROL, and RAL to elucidate retinoid homeostasis in disease states such as Alzheimer's disease, type 2 diabetes, obesity, and cancer. PMID- 18410740 TI - Communications between bone cells and hematopoietic stem cells. AB - The skeletal system, while characterized by a hard tissue component, is in fact an extraordinarily dynamic system, with disparate functions ranging from structural support, movement and locomotion and soft-organ protection, to the maintenance of calcium homeostasis. Amongst these functions, it has long been known that mammalian bones house definitive hematopoiesis. In fact, several data demonstrate that the bone microenvironment provides essential regulatory cues to the hematopoietic system. In particular, interactions between the bone-forming cells, or osteoblasts, and the most primitive Hematopoietic Stem Cells (HSC) have recently been defined. This review will focus mainly on the role of osteoblasts as HSC regulatory cells, discussing the signaling mechanisms and molecules currently thought to be involved in their modulation of HSC behavior. We will then review additional cellular components of the HSC niche, including endothelial cells and osteoclasts. Finally, we will discuss the potential clinical implications of our emerging understanding of the complex HSC microenvironment. PMID- 18410741 TI - The dynamic interplay between osteoclasts and the immune system. AB - Investigation into arthritis, as well as numerous bone phenotypes found in mice lacking immune-related genes, has highlighted the importance of the interplay between the bone and immune systems, which has led to the emergence and evolution of the field of osteoimmunology. RANKL stimulates osteoclastogenesis through nuclear factor of activated T cells (NFAT) c1, which is also a crucial regulator of immunity. In rheumatoid arthritis, bone destruction is caused by the enhanced activity of osteoclasts, which is mainly dependent on interleukin-17-producing helper T cells (T(H)17). The scope of osteoimmunology has been extended to encompass a wide range of molecular and cellular interactions. The framework of osteoimmunology will provide a scientific basis for future therapeutic approaches to diseases related to both of these systems. PMID- 18410743 TI - Spectral monitoring of photoirradiated skin lipids: MS and IR approaches. AB - An investigation of the effects of UV(A) irradiation on the stratum corneum lipids was carried out in vitro on films. The modifications of their conformational order were studied by FTIR and the formation of new entities was detected by mass spectroscopy. The results show not only differences in behaviour of the three lipid classes (fatty acids (FA), ceramides (CER), and cholesterol), but also variation within a class, depending on the molecules structure. Upon UV(A) irradiation, beta scission occurs on all the components, saturated and unsaturated. Moreover, unsaturated FA or CER having a double bond on their FA moiety may become saturated or may be transformed into their free radical form. Unsaturated FA are more sensitive to UV(A) and lead more easily to oxygenated components than unsaturated CER. The chemical effects are irradiation dose dependent but do not deeply influence the supramolecular organisation of these lipids. The global conformation of the lipids stays in an orthorhombic state, a decrease of the packing density however is observed. PMID- 18410744 TI - The holistic approach to teach laboratory medicine. AB - Laboratory diagnostics has undergone relevant changes in organization and complexity, providing new opportunities and risks. The skill and expertise required for laboratory professionals to fulfil the need of a continuously evolving scenario in diagnostics embrace now a variety of scientific, managerial and organizational issues. Translating into clinical practice new insights from basic science, particularly regarding omics, requires construction of a new and complex core curriculum for laboratory professionals. In addition, the integration of different areas of diagnostics within the same laboratory service is expected to improve efficiency and effectiveness. PMID- 18410742 TI - Regulation of bone remodeling by the central and peripheral nervous system. AB - The homeostatic nature of bone remodeling has become a notion further supported lately by the demonstration that neuropeptides and their receptors regulate osteoblast and osteoclast function in vivo. Following initial studies reporting the presence of nerves and nerve-derived products within the bone microenvironment and the expression of receptors for these neuropeptides in bone cells, new experimental and mechanistic evidence based on in vivo murine genetic and pharmacologic models recently demonstrated that inputs from the central and peripheral nervous system feed into the already complex regulatory machinery controlling bone remodeling. The function of a number of "osteo-neuromediators" has been characterized, including norepinephrine and the beta2-adrenergic receptor, Neuropeptide Y and the Y1 and Y2 receptors, endocannabinoids and the CB1 and CB2 receptors, as well as dopamine, serotonin and their receptors and transporters, Calcitonin gene-related peptide, and neuronal NOS. This new body of evidence suggests that neurons in the central nervous system integrate clues from the internal and external milieux, such as energy homeostasis, glycemia or reproductive signals, with the regulation of bone remodeling. The next major tasks in this new area of bone biology will be to understand, at the molecular level, the mechanisms by which common central neural systems regulate and integrate these major physiological functions, the relative importance of the central and peripheral actions of neuropeptides present in both compartments and their relationship, and how bone cells signal back to central centers, because the definition of a homeostatic function implies the existence of feedback signals. Together, these findings shed a new light on the complexity of the mechanisms regulating bone remodeling and uncovered new potential therapeutic strategies for the design of bone anabolic treatments. This review summarizes the latest advances in this area, focusing on investigations based on in vivo animal studies. PMID- 18410745 TI - Meeting the curriculum needs for different career paths in Laboratory Medicine. AB - There are a number of career paths in Laboratory Medicine and several clinical practice models for the discipline. This article summarizes the state of current training at the medical student and residency/post-graduate levels, emphasizing practice in the U.S., and the challenges of education in the discipline to meet the needs of diverse career paths. Data regarding effectiveness of current pedagogical approaches are discussed along with a brief review of evolving didactic methodologies. The recently published curriculum in Laboratory Medicine (Clinical Pathology) by the Academy of Clinical Laboratory Physicians and Scientists is reviewed, including its major emphases and the importance of competency assessment. Finally, the future of Laboratory Medicine and Pathology and the need to train for that future is expanded upon. PMID- 18410746 TI - NeoR6 inhibits HIV-1-CXCR4 interaction without affecting CXCL12 chemotaxis activity. AB - Aminoglycoside-arginine conjugates (AACs) are multi-target HIV-1 inhibitors. The most potent AAC is neomycin hexa-arginine conjugate, NeoR6. We here demonstrate that NeoR6 interacts with CXCR4 without affecting CXCL12-CXCR4 ordinary chemotaxis activity or loss of CXCR4 cell surface expression. Importantly, NeoR6 alone does not affect cell migration, indicating that NeoR6 interacts with CXCR4 at a distinct site that is important for HIV-1 entry and mAb 12G5 binding, but not to CXCL12 binding or signaling sites. This is further supported by our modeling studies, showing that NeoR6 and CXCL12 bind to two distinct sites on CXCR4, in contrast with other CXCR4 inhibitors, e.g. T140 and AMD3100. This complementary utilization of chemical, biology, and computation analysis provides a powerful approach for designing anti-HIV-1 drugs without interfering with the natural function of CXCL12/CXCR4 binding. PMID- 18410747 TI - Thioredoxins of a parasitic nematode: comparison of the 16- and 12-kDA thioredoxins from Haemonchus contortus. AB - Thioredoxins are a family of small proteins conserved through evolution, which are essential for the maintenance of cellular homeostasis. The "classic" thioredoxin, identified in most species, is a 12-kDa protein with a Cys-Pro-Gly Cys (CPGC) active site. However, in nematodes a larger protein, 16 kDa, with a Cys-Pro-Pro-Cys (CPPC) active site was identified. We report that in the parasitic nematode Haemonchus contortus, both the 12-kDa (HcTrx1) and the 16-kDa (HcTrx3) species are expressed through the life cycle. However, the HcTrx3 is expressed at higher concentrations. Recombinant HcTrx1 and HcTrx3 were produced and both reduced insulin at a rate similar to that observed with ovine (host) and Escherichia coli thioredoxins and both were regenerated by a mammalian thioredoxin reductase, demonstrating that they have similar thioredoxin activity. Unlike mammalian thioredoxins, both proteins were able to reduce oxidised glutathione and hydrogen peroxide. This suggests essential roles for these proteins in response to oxidative stress and the host immune attack. Analysis of ivermectin-resistant H. contortus showed that expression of both genes were increased in a drug-resistant strain relative to a sensitive strain. Involvement in drug resistance identifies these thioredoxin proteins as potential drug targets for parasite control. PMID- 18410756 TI - Creating an academic culture during residency training. PMID- 18410757 TI - Which comes first? The obesity or the insulin? The behavior or the biochemistry? PMID- 18410758 TI - A boost to the study of insulin secretion in children and adolescents. PMID- 18410759 TI - PHIShing for clarity in the management of apparent life-threatening events. PMID- 18410760 TI - From liquid to air: breathing after birth. PMID- 18410761 TI - Insulin resistance, hyperinsulinemia, and energy intake in overweight children. AB - OBJECTIVE: To examine the relationship between energy intake during a buffet meal and indexes of insulin dynamics in overweight children. STUDY DESIGN: Ninety-five nondiabetic, overweight (body mass index > or = 95th percentile) children (age 10.3 +/- 1.4 years) selected lunch from a 9835-kcal buffet eaten ad libitum after an overnight fast. The associations between energy intake and measures of insulin dynamics, in the postabsorptive state and during a 2-hour hyperglycemic clamp, were determined. Covariates in the statistical model included race, sex, skeletal age, fat-free mass, fat mass, socioeconomic status, and number of foods in the buffet rated as acceptable. RESULTS: Energy intake was positively associated with the fasting homeostasis model assessment for insulin resistance index (beta = 0.24, P = .042), fasting insulin/glucose ratio (beta = 0.24, P = .044), first phase insulin (beta = 0.23, P = .032), and first-phase C-peptide (beta = 0.21, P = .046); energy intake was negatively associated with clamp-derived insulin sensitivity (beta = -0.29, P = .042). Each 10% decrease in clamp-derived insulin sensitivity predicted a 27-kcal greater energy intake. CONCLUSIONS: Insulin resistance and hyperinsulinemia are associated with greater energy intake after an overnight fast in overweight children. These associations suggest mechanisms whereby insulin resistance may contribute to excessive weight gain in children. PMID- 18410762 TI - Measures of beta-cell function during the oral glucose tolerance test, liquid mixed-meal test, and hyperglycemic clamp test. AB - OBJECTIVE: To evaluate clinically useful measures of beta-cell function derived from the oral glucose tolerance test (OGTT) or mixed-meal (ie, Boost) tolerance test to assess insulin secretion in comparison with the gold standard, the hyperglycemic clamp (Hyper-C) test. STUDY DESIGN: We hypothesized that OGTT/Boost derived measures are useful estimates of beta-cell function and correlate well with insulin secretion measured by the Hyper-C test. This study was designed to assess the correlation between the ratio of the early incremental insulin/glucose responses at 15 and 30 minutes (DeltaI(15)/DeltaG(15) and DeltaI(30)/DeltaG(30)) of the OGTT and the Boost test with insulin secretion measured during the Hyper-C test (225 mg/dL). The same indices were evaluated using C-peptide. A total of 26 children (14 males, 12 females; mean age, 9.9 +/- 0.2 years; mean body mass index = 22.1 +/- 1.2 kg/m(2)) underwent a 2-hour Hyper-C test (225 mg/dL) and 3-hour OGTT and Boost tests with measurements of glucose, insulin, and C-peptide. RESULTS: Correlations between Hyper-C- and OGTT-derived measures of insulin secretion were stronger for the 15-minute index than for the 30-minute index of insulin secretion and stronger for C-peptide levels than for insulin levels (r = .7, P < .001 for first-phase C-peptide vs both OGTT and Boost, DeltaC(15)/DeltaG(15)). CONCLUSIONS: In children with normal glucose tolerance, C peptide rather than insulin level measured after 15 minutes of the OGTT or Boost test provides a reliable estimate of beta-cell function that correlates well with Hyper-C-derived insulin secretion. PMID- 18410763 TI - Dermatological complications of continuous subcutaneous insulin infusion in children and adolescents. AB - OBJECTIVES: To describe the dermatological changes associated with continuous subcutaneous insulin infusion (CSII) therapy in youth with type 1 diabetes mellitus (T1D). To assess their association with duration of CSII, age, adiposity, HbA1(c), insulin dose, insulin brand, infusion set or site. STUDY DESIGN: We conducted a cross-sectional study of 50 consecutive patients with T1D who were using CSII for >6 months (26 female; age, 13.3 +/- 3.5 years [mean +/- SD]; CSII duration, 2.8 +/- 1.7 years; HbA1(c), 7.7% +/- 1.1%). A grading scale was devised. Ultrasound scanning was performed in 8 subjects. RESULTS: The mean (+/-SD) severity score was 6.3 +/- 3.5 (range, 0-14; maximum possible, 69). Most common were scars <3 mm diameter (94%), erythema not associated with nodules (66%), subcutaneous nodules (62%), and lipohypertrophy (42%). There was a significant negative correlation between severity score and body mass index z score (r = -0.3, P = .039), but no correlation with HbA1(c), insulin brand or site. Infusion sets inserted at 90 degrees were associated with lower scores (P = .03). Less than 5% of patients and parents considered stopping CSII because of skin concerns. Ultrasound scanning results of CSII sites revealed mild increased echogenicity of the dermis and hypodermis. CONCLUSIONS: Dermatological changes were frequent, with increased severity associated with lower adiposity. These complications were not associated with glycemic control, nor did they prompt most to consider stopping CSII. PMID- 18410764 TI - Variation in inpatient resource utilization and management of apparent life threatening events. AB - OBJECTIVE: To report national variations in diagnostic approaches to apparent life-threatening events (ALTEs) and resource utilization. STUDY DESIGN: Using the Pediatric Health Information System, we studied children who were age 3 days to 5 months at admission and were discharged with an International Classification of Diseases, Ninth Revision (ICD-9) code potentially identifiable as ALTE. Multiple analysis of variance was used to determine whether the variances in adjusted charges, length of stay (LOS), and diagnostic studies were hospital-related after controlling for other covariates. Logistic regression was used to study the association of readmission rates with discharge diagnosis and specific diagnostic studies. RESULTS: The study group comprised 12,067 patients, with a mean LOS of 4.4 days (standard deviation +/- 5.6 days) and mean adjusted charges of $15,567 ($28,510) per admission. The mean in-hospital mortality rate was 0.56% (n = 68), and the rate of 30-day readmission was 2.5%. The most common discharge diagnoses were gastroesophageal reflux 36.9% (48.3%) and lower respiratory tract infection 30.8% (46.2%). Mean LOS, total adjusted charges, and use of diagnostic studies varied considerably across hospitals, and hospital-level differences were a significant contributor to the variance of these outcomes after controlling for covariates (P < .001). There was an increased likelihood of readmission for patients discharged with a diagnosis of cardiovascular disorders (odds ratio [OR] = 1.68; 95% confidence interval [CI] = 1.30 to 2.16) and gastroesophageal reflux (OR = 1.32; 95% CI = 1.03 to 1.69) compared with other discharge diagnoses. CONCLUSIONS: There is considerable hospital-based variation in care for patients hospitalized for conditions potentially identifiable as ALTE, particularly in the evaluation and diagnosis of gastroesophageal reflux, which may contribute to adverse clinical and financial outcomes. An evidence-based national standard of care for ALTE is needed, as are multi-institutional initiatives to study different diagnostic and management strategies and their effect on patient outcomes. PMID- 18410765 TI - Extreme and conventional cardiorespiratory events and epidemiologic risk factors for SIDS. AB - OBJECTIVE: To test the hypotheses that there is a lack of correlation between extreme events and epidemiologic risk factors for sudden infant death syndrome (SIDS), and if conventional events are normal, their numbers should increase once a circadian decrease in breathing rate is established. In addition, the number of events should decrease with maternal smoking. STUDY DESIGN: Three outcome variables were derived from the Collaborative Home Infant Monitoring Evaluation (CHIME) of 1082 infants: (1) at least 1 extreme event lasting > or = 30 seconds, (2) at least 1 conventional event lasting > or = 20 seconds, and (3) being part of the 50% of infants with the most events. RESULTS: Multivariate logistic regression analyses found that extreme events were not statistically associated with any known SIDS risk factors and occurred less often during the early morning. Healthy term infants had significantly fewer of these events compared with preterm infants, subsequent siblings of infants with SIDS, and infants with an apparent life-threatening event, a finding that was not evident after 43 weeks (3 weeks postterm). Conventional events increased during the night, whereas maternal smoking was associated with a decrease in conventional events. Apneic episodes persisting for > or = 40 seconds occurred in 1.8% of the infants. CONCLUSIONS: Extreme events are associated with immaturity and do not seem to be immediate precursors of or causally related to SIDS. PMID- 18410766 TI - Does placental inflammation relate to brain lesions and volume in preterm infants? AB - OBJECTIVES: To evaluate the association between histologic inflammation of placenta and brain findings in ultrasound examinations and regional brain volumes in magnetic resonance imaging in very-low-birth-weight (VLBW) or in very preterm infants. STUDY DESIGN: VLBW or very preterm infants (n = 121) were categorized into 3 groups according to the most pathologic brain finding on ultrasound examinations until term. The brain magnetic resonance imaging performed at term was analyzed for regional brain volumes. The placentas were analyzed for histologic inflammatory findings. RESULTS: Histologic chorioamnionitis on the fetal side correlated to brain lesions in univariate but not in multivariate analyses. Low gestational age was the only significant risk factor for brain lesions in multivariate analysis (P < .0001). Histologic chorioamnionitis was not associated with brain volumes in multivariate analyses. Female sex, low gestational age, and low birth weight z score correlated to smaller volumes in total brain tissue (P = .001, P = .0002, P < .0001, respectively) and cerebellum (P = .047, P = .003, P = .001, respectively). In addition, low gestational age and low-birth-weight z score correlated to a smaller combined volume of basal ganglia and thalami (P = .0002). CONCLUSIONS: Placental inflammation does not appear to correlate to brain lesions or smaller regional brain volumes in VLBW or in very preterm infants at term age. PMID- 18410767 TI - Neurodevelopmental outcome of preterm infants with severe intraventricular hemorrhage and therapy for post-hemorrhagic ventricular dilatation. AB - OBJECTIVE: To evaluate the neurodevelopmental outcome of preterm infants with a grade III or IV hemorrhage and to assess the effect of routine low-threshold therapy of post-hemorrhagic ventricular dilatation (PHVD) on neurodevelopmental outcome. STUDY DESIGN: Of the 214 preterm infants (< or = 34 weeks gestational age), 94 (44%) had a grade III intraventricular hemorrhage (IVH), and 120 (56%) had a grade IV hemorrhage. We evaluated the natural evolution of IVH, the need for intervention for PHVD, and neurodevelopmental outcome at 24 months corrected age. RESULTS: PHVD developed significantly more often in the surviving infants with a grade III hemorrhage (53/68, 78%) than in infants with a grade IV hemorrhage (40/76, 53%; P = .002). Intervention for PHVD was required significantly more often in the grade III group, than in the grade IV group (P < .001). In the grade III group, cerebral palsy developed in 5 of the 68 surviving infants (7.4%), compared with 37 of the 76 infants (48.7%) with a grade IV hemorrhage (P < .001). The mean developmental quotient (DQ) in the grade III group was 99, and in the grade IV-group it was 95 at 24 months corrected age. CONCLUSIONS: Short-term neurodevelopmental outcome of preterm infants with grade III or IV hemorrhage was better than reported earlier. Requiring intervention for PHVD only had a negative effect on DQ in infants with a grade IV hemorrhage. Infants with cerebral palsy had significantly lower DQs, irrespective of the severity of IVH. PMID- 18410769 TI - Neonatal vaccination with an acellular pertussis vaccine accelerates the acquisition of pertussis antibodies in infants. AB - OBJECTIVES: Because young infants are at highest risk of pertussis complications, this study assessed whether neonatal acellular pertussis (aP) vaccination could provide earlier immunity. STUDY DESIGN: Neonates (n = 121) were randomly assigned (1:1) to receive either aP or hepatitis B vaccine (HBV) (controls) vaccine at birth, followed by vaccination with DTaP-HBV-IPV/Hib at 2, 4 and 6 months. Immune responses were measured. Reactogenicity was assessed for 7 days after each dose. RESULTS: The aP birth dose was followed by few adverse events. Reactogenicity of subsequent vaccine doses did not differ between groups. Seven serious adverse events were reported from each group; none were related to the study vaccines. At 3 months of age, vaccination with aP at birth had induced significantly higher antibody responses to the 3 pertussis antigens compared with controls. At 7 months, geometric mean/concentrations of antibodies against pertussis antigens were similar in both groups, and all subjects had reached "seroprotective" antibody concentrations against diphtheria, tetanus, and poliovirus types 1, 2, and 3. Geometric mean/concentrations of antibodies to haemophilus influenzae type b (Hib) and HBV were significantly lower in the aP group. CONCLUSIONS: Early neonatal immunization with aP was safe, well tolerated, and resulted in earlier antibody responses, seen after the first dose of a DTaP combination vaccine. Birth dose of aP did not induce immunologic tolerance to pertussis antigens but appear to dampen responses to Hib and HBV vaccines. PMID- 18410770 TI - Respiratory viruses in laryngeal croup of young children. AB - OBJECTIVES: To determine the viral cause of laryngeal croup by use of highly sensitive methods, and including recently recognized viruses in the analysis. STUDY DESIGN: One hundred forty-four consecutive children with hoarse voice and inspiratory stridor attending the emergency department were enrolled. Age- and season-matched children presenting with a wheezing illness served as control subjects (n = 76). Nasopharyngeal swabs were analyzed by polymerase chain reaction for rhinovirus and enterovirus, coronavirus, respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza A and B virus, human bocavirus, human metapneumovirus, adenovirus, and Mycoplasma pneumoniae. RESULTS: Virus infection was documented in 80% of patients with croup and 71% of control subjects. Children with croup had significantly more positive test results for PIV 1 and 2 (31% vs 4% and 6% vs 0%, respectively) and significantly fewer positive test results for RSV (15% vs 28%) than wheezing children. Rhinoviruses and enteroviruses were present equally in both groups (21% vs 25%). There was no significant difference in the frequency of influenza A virus or human bocavirus. Few subjects with adenovirus or M. pneumoniae were detected. CONCLUSION: Acute laryngeal croup is most often associated with PIV, RSV, rhinovirus, and enterovirus. Rhinovirus and enterovirus appeared equally often in croup and in wheezing illness. During late fall, they were found in 39% and 40%, respectively, of the tested samples. PMID- 18410771 TI - Lack of piped water and sewage services is associated with pediatric lower respiratory tract infection in Alaska. AB - OBJECTIVES: To determine the association between the high incidence of lower respiratory tract infection (LRI) documented among young Alaskan children and the absence of modern water service (in-home piped water/septic system or water delivered by closed haul truck) found commonly in rural Alaskan communities. STUDY DESIGN: A community-level analysis was performed of all 108 Alaskan communities with at least 15 children <2 years of age enrolled in Medicaid during 1998-2003. Community LRI incidence rates were determined from a Medicaid database with standard LRI billing codes. Potentially confounding community-level demographic variables were obtained, as was availability of water service. RESULTS: During linear regression analysis, the percentage of households with modern water service in a community predicted community-level outpatient (beta = 0.53; P < .001) and inpatient (beta = -0.15; P = .088) LRI incidence rates when controlling for the degree of household crowding, unemployment, adult education, tobacco cigarette use, wood stove use, and poverty. Modest improvements in water service delivery were not shown to be associated with changes in LRI burden. CONCLUSIONS: Lack of modern water service in Alaska is associated with high pediatric LRI incidence. These communities should receive modern water service, but this intervention alone may not dramatically reduce LRI burden. PMID- 18410772 TI - Infectious, malignant, and autoimmune complications in pediatric heart transplant recipients. AB - OBJECTIVE: To review clinical courses of pediatric heart transplant survivors after 5 years from transplantation for infections, lymphoproliferative, and autoimmune diseases. STUDY DESIGN: A total of 71 patients were examined in 2 groups, infant recipients (underwent transplant <1 year of age, n = 38) and older recipients (underwent transplant >1 year, n = 33). All patients received comparable immunosuppression. Calculated occurrence rates were reported as means per 10 years of follow-up with SEs. Differences were examined by using Poisson regression. RESULTS: Infant recipients had significantly higher (P < .001) occurrence rates of severe (mean, 2.04 +/- 0.5) and chronic infections (mean, 4.58 +/- 0.67) compared with older recipients (means, 0.37 +/- 0.19 and 1.87 +/- 0.70, respectively). Types of infections were similar to those in the general population with extremely rare opportunistic infections; however, they were more severe and resistant to treatment. Autoimmune disorders occurred at a frequency comparable with lymphoproliferative diseases and were observed in 7 of 38 infants (18%). Most common were autoimmune cytopenias. CONCLUSIONS: Infant heart transplant recipients who survive in the long term have higher occurrence rates of infections compared with older recipients. Autoimmune disorders are a previously unrecognized morbidity in pediatric heart transplantation. PMID- 18410773 TI - Anthropometric and behavioral patterns associated with weight maintenance after an obesity treatment in adolescents. AB - OBJECTIVE: To identify anthropometric and behavioral characteristics associated with weight maintenance after an obesity treatment. STUDY DESIGN: Adolescents (n = 72) enrolled in a 9-month obesity treatment were observed 1 and 2 years after discharge. Two groups, "successful" versus "limited or no success," were created on the basis of the differences in body mass index (BMI) z-score between inclusion and end of follow-up. Anthropometric and behavioral characteristics were compared between groups. RESULTS: Both groups showed a decrease in BMI z score between inclusion and end of follow-up, 2.09 +/- 0.68 SD for the successful group and 0.65 +/- 0.43 SD for the group with limited or no success. Groups did not differ during treatment for any of the anthropometric characteristics considered, whereas differences clearly appeared 1 year after treatment and generally stabilized during the second year. Later adiposity rebound, trend for lower BMI in the mother, and, during follow-up, lower total energy intake, more energy at breakfast, and less snacking and television were recorded in the successful group. CONCLUSIONS: Weight loss maintenance can neither be predicted with anthropometry during treatment nor with behavioral characteristics at inclusion, but can be estimated 1 year after discharge. Early life factors should also be taken into account for predicting treatment outcome. PMID- 18410774 TI - Double-blind placebo-controlled trial of amitriptyline for the treatment of irritable bowel syndrome in adolescents. AB - OBJECTIVES: To determine the efficacy of amitriptyline (AMI) in treating irritable bowel syndrome (IBS) in adolescents. STUDY DESIGN: Adolescents 12 to 18 years with newly diagnosed IBS were surveyed with a symptom checklist, pain rating scale, visual analog scale, and IBS quality of life (QOL) questionnaire. Subjects were randomized in a double-blinded fashion to receive AMI or placebo, and again completed surveys at 2, 6, 10, and 13 weeks. RESULTS: Thirty-three patients (24 female) were enrolled. Patients receiving AMI were more likely to experience improvement from baseline in overall QOL at 6, 10, and 13 weeks (P = .019, .004, and .013). Patients receiving AMI were also more likely to experience a reduction in IBS-associated diarrhea at 6 and 10 weeks (P = .029 for both), a reduction in periumbilical pain at 10 weeks (P = .018), and a reduction in right lower quadrant pain at 6, 10, and 13 weeks (P = .014, .039, and .004). CONCLUSION: AMI significantly improves overall QOL in adolescents with IBS and should be a therapeutic option for adolescents with this disorder. PMID- 18410776 TI - High growth rate of girls with precocious puberty exposed to estrogenic mycotoxins. AB - OBJECTIVE: To test the hypothesis that human puberty timing can be advanced by environmental estrogen exposure. STUDY DESIGN: We analyzed serum mycoestrogen contamination via high-performance liquid chromatography (HPLC) in 32 girls affected by central precocious puberty (CPP) and in 31 healthy female control subjects. All 32 patients received triptorelin (TR) for more than 12 months after diagnosis. RESULTS: Increased serum levels of zearalenone (ZEA; 933.7 +/- 200.3 pg/mL; 95% CI, 723.5-1143.9) and of its congener alpha-zearalenol (106.5 +/- 1.9 pg/mL; 95% CI, 104.5-108.5) contaminated 6 girls with CPP, who were from a bounded Tuscany area. At diagnosis, ZEA levels correlated with patient height (r = 0.906, P < .05) and weight (r = 0.887, P < .05), but not with bone age. In patients who were mycotoxin-positive, height (F = 4.192; P < .01), weight (F = 3.915; P < .01), and height velocity (F = 2.777, P < .05) were higher than patients who were mycotoxin-negative during 12-months TR treatment. Height correlated with weight both in patients who were mycotoxin-positive (r = 0.986, P < .001) and in patients who were mycotoxin-negative (r = 0.994, P < .001). Body mass index, bone age, and gonadal secretion was not different in patient groups before and during TR treatment (P > .05). CONCLUSIONS: Mycoestrogenic zearalenone is suspected to be a triggering factor for CPP development in girls. Because of its chemical resemblance to some anabolic agents used in animal breeding, ZEA may also represent a growth promoter in exposed patients. PMID- 18410777 TI - Dose-response relationships between iron deficiency with or without anemia and infant social-emotional behavior. AB - OBJECTIVE: To assess dose-response relationships between severity of iron deficiency (ID) and infant social-emotional behavior. STUDY DESIGN: The study group was a cohort of 9- to 10-month-old African-American infants (n = 77 with final iron status classification). The infants were given oral iron for 3 months. Social-emotional outcomes included mother and examiner ratings at 9 and 12 months and quantitative behavioral coding from videotape at 12 months. General linear model analyses tested for linear effects of iron status group (ordered from worst to best: iron-deficient anemia [IDA], nonanemic iron-deficient [NA ID], iron sufficient [IS]) and determined thresholds for effects. RESULTS: There were significant (P <.05) linear effects of poorer iron status for shyness (increasing, maternal rating), orientation-engagement, and soothability (decreasing, examiner ratings), and the following quantitatively coded behaviors: positive affect (decreasing) and latencies to engage with the examiner (increasing) and move away from the examiner (decreasing). The threshold for all but 1 effect was ID with or without anemia versus IS. CONCLUSIONS: Infant social emotional behavior appears to be adversely affected by ID with or without anemia. ID without anemia is not detected by common screening procedures and is more widespread than IDA. Infant social-emotional behavior can profoundly influence the care-giving environment, with repercussions for overall development. PMID- 18410778 TI - Longitudinal analysis of resting energy expenditure in patients with cystic fibrosis. AB - OBJECTIVE: To assess whether elevated resting energy expenditure (REE) in female patients with cystic fibrosis (CF) persists longitudinally during late childhood and puberty. STUDY DESIGN: REE and body composition were measured 3 times in 2 years in 86 children with CF. Pubertal status, bacterial colonization, liver disease, pancreatic and pulmonary function, and genotype were determined, and linear mixed model analyses were used to determine predictors and changes in REE longitudinally. RESULTS: REE did not change with time allowing for fat free mass, pancreatic insufficiency (PI), or severe mutations. Pulmonary function and liver disease were not significant predictors of REE. Percentage predicted REE compared with control data was higher (P = .002) in female patients with CF (109.5%) and lower in male patients with CF (104%) and persisted with time. In post-menarchal female patients with CF, REE adjusted for fat free mass was 366 kJ/d lower than in pre-menarchal female patients, but still 112% predicted. CONCLUSIONS: This longitudinal study demonstrates that REE is elevated in patients with CF with PI and severe mutations. The elevation of percentage predicted REE was greater in female patients than male patients and persisted for 2 years, and during pubertal maturation, independent of pulmonary and liver disease. These results highlight the need for a high-energy diet throughout childhood and adolescence, particularly in female patients with PI. PMID- 18410779 TI - Environmental tobacco smoke and interleukin 4 polymorphism (C-589T) gene: environment interaction increases risk of wheezing in African-American infants. AB - OBJECTIVES: To determine whether infants exposed to environmental tobacco smoke (ETS) having the interleukin 4 (IL-4) or interleukin 13 (IL-13) gene polymorphisms were at increased risk of wheezing. STUDY DESIGN: A birth cohort of 758 infants was evaluated annually by a questionnaire, physical examination, and skin prick testing. DNA samples from 560 children were genotyped for IL-4 C-589T and IL-13 C-1112T. The relationship of ETS exposure and genotype with the outcome of wheezing was analyzed. RESULTS: At the time of evaluation, mean age was 13.4 +/- 2.2 months. The prevalence of sensitization was 29%, and wheezing without a cold was 26.2%. The interaction of ETS exposure and the CT/TT genotypes for IL-4 C-589T showed a significant association with wheezing (odds ratio: 10.84; 95% confidence interval: 1.12-104.64, P = .04) in African-American infants. CONCLUSIONS: In African-American infants with a family history of atopy, the interaction of ETS and IL-4 C-589T demonstrated a 10-fold risk associated with wheezing without a cold. PMID- 18410780 TI - Effect of ascertainment and genetic features on the phenotype of Klinefelter syndrome. AB - OBJECTIVE: To describe the Klinefelter Syndrome (KS) phenotype during childhood in a large cohort. STUDY DESIGN: Clinical assessment, measurement of hormonal indices of testicular function, and parent of origin of extra X chromosome were assessed in a cross-sectional study of 55 boys with KS, aged 2.0 to 14.6 years, at an outpatient center. RESULTS: Mean height and body mass index SD scores (SDS +/- SD) were 0.9 +/- 1.3 and 0.4 +/- 1.4, respectively. Mean penile length and testicular volume SDS were -0.5 +/- 0.9 and -0.9 +/- 1.4. Testosterone levels were in the lowest quartile of normal in 66% of the cohort. Other features included clinodactyly (74%), hypertelorism (69%), elbow dysplasia (36%), high arched palate (37%), hypotonia (76%), and requirement for speech therapy (69%). Features were similar in boys in whom the diagnosis was made prenatally versus boys in whom the diagnosis was made postnatally. There was no evidence for a phenotypic effect of parent of origin of the extra X chromosome. CONCLUSIONS: Boys with KS commonly have reduced penile length and small testes in childhood. The phenotype in boys with KS does not differ according to ascertainment or origin of the extra X chromosome. Boys with KS may be identified before puberty by tall stature, relatively decreased penile length, clinodactyly, hypotonia, and requirement for speech therapy. PMID- 18410781 TI - Enzyme replacement therapy for mucopolysaccharidoses: opinions of patients and families. AB - OBJECTIVES: To assess the opinions of individuals with mucopolysaccharidoses (MPS) and their parents regarding the use of enzyme replacement therapy (ERT). STUDY DESIGN: A validated questionnaire, including hypothetical clinical scenarios about ERT for MPS, was distributed to members of MPS support groups in the United States and Australia. RESULTS: The questionnaire was completed by 249 MPS support group members. Overall, 92% were in favor of ERT where MPS causes severe physical problems but does not affect intellect, and 69% were in favor of ERT where the physical limitations are mild and intellect is spared. Only 47% were in favor of ERT where severe physical and intellectual problems are well established; however, 77% were in favor of ERT in this situation if treatment begun early prolongs life and improves quality of life. CONCLUSION: Most respondents were in favor of ERT for MPS, even where it would not alter the intellectual deterioration. The medical community has a responsibility to advocate for their patients in situations where ERT is appropriate and recognize the economic burden and "family function burden" ERT can incur. PMID- 18410782 TI - Urinary loss of erythropoietin after intravenous versus subcutaneous epoetin-beta in preterm infants. AB - Hematopoietic and non-hematopoietic effects of recombinant erythropoietin (Epo) given to preterm infants are controversially discussed. Because renal loss of Epo was significantly higher after intravenous versus subcutaneous Epoetin-beta administration, we suggest a reconsideration of whether subcutaneous recombinant Epo is more efficient and safer because of lower peaks of circulating Epo. PMID- 18410783 TI - Neonatal vitamin B12 deficiency secondary to maternal subclinical pernicious anemia: identification by expanded newborn screening. AB - A neonate with elevated propionylcarnitine on the newborn screen was found to have methylmalonic acidemia due to vitamin B(12) deficiency. The mother was also vitamin B(12)-deficient. This case illustrates the utility of expanded newborn screening for detection of vitamin B(12) deficiency, allowing prompt treatment and prevention of potential sequelae. PMID- 18410785 TI - Screening for domestic violence and childhood exposure in families seeking care at an urban pediatric clinic. AB - In a survey of 133 caregivers in a pediatric clinic, 30 women (23%) disclosed domestic violence, with 2 reporting coercive control but not physical violence. Seventeen women stated that a child had been exposed as well. Domestic violence is not a "private" adult problem; further study of an appropriate pediatric-based screener is needed. PMID- 18410786 TI - Coarctation of the aorta. PMID- 18410787 TI - Bupropion assists with tobacco cessation in adolescents but relapse is high. PMID- 18410788 TI - Exercise training increases physical fitness for children with cerebral palsy. PMID- 18410789 TI - Honey improves cough in children compared to no treatment. PMID- 18410790 TI - Use of caffeine for apnea of prematurity also has long-term neurodevelopmental benefits. PMID- 18410791 TI - Tonsillectomy for periodic fever, aphthous stomatitis, pharyngitis, and adenitis syndrome is not always successful. PMID- 18410792 TI - The role of tonsillectomy in management of periodic fever, aphthous stomatitis, pharyngitis, and adenopathy: Unanswered questions. PMID- 18410794 TI - Cellular mechanisms of resistance to anthracyclines and taxanes in cancer: intrinsic and acquired. AB - Taxanes and anthracyclines are two of the most potent and broadly effective classes of chemotherapeutic agents. However, resistance to these agents is common and significantly limits their potential. As such, there is a great need to understand the mechanisms underlying de novo and acquired resistance to these agents. Beyond the resistance barrier lies even greater potential to significantly alter the natural course of human cancer. This review discusses what we currently understand about the mechanisms of resistance to taxanes and anthracyclines. Preclinical models suggest a role for ATP-binding cassette transporters, tubulin isoforms, microtubule-associated proteins, tubulin gene mutations, and mitotic checkpoint signaling proteins in resistance to taxanes. Preclinical models also suggest that drug transport proteins, antioxidant defenses, apoptotic signaling, and topoisomerase modulation may mediate anthracycline resistance. Many of these hypotheses remain untested in appropriately designed clinical studies, but limited clinical evidence will be reviewed. Epothilones represent a novel class of non-taxane microtubule stabilizing agents with distinct drug-resistance profiles. Potential mechanisms behind these differences and their potential role in the treatment of both taxane and anthracycline-refractory patients are discussed. PMID- 18410795 TI - Preclinical investigations with epothilones in breast cancer models. AB - The epothilones constitute a novel class of microtubule inhibitors that act like the taxanes by hyperstabilizing tubulin polymerization, thus disrupting functioning of the mitotic spindle. Natural epothilones produced by myxobacteria, and second- or third-generation partially or fully synthesized analogs, have been explored as cancer chemotherapy agents to replace or follow the taxanes. For those epothilones that have gone on to clinical development (epothilone B, ixabepilone, BMS-310705, ZK-EPO, KOS-862, and KOS-1584), preclinical investigations in breast cancer models are reviewed. All of these epothilones improve upon the cytotoxic activity of paclitaxel in various human breast cancer cell lines in vitro, but are also highly active in lines that are resistant to paclitaxel. Comparable antitumor activity has been demonstrated against nude mouse xenografts of paclitaxel-sensitive and -resistant breast cancer lines. Additionally, some analogs have reduced toxicity or increased water solubility that may permit oral administration, while others with enhanced tissue penetration show promise in animal models of breast cancer brain or bone metastasis and may provide benefits in patients with poor-prognosis advanced breast cancer. PMID- 18410796 TI - Clinical studies with epothilones for the treatment of metastatic breast cancer. AB - Standard cytotoxic chemotherapy of locally advanced or metastatic breast cancer includes the microtubule-stabilizing taxanes, but like other cytotoxic drugs their effectiveness is compromised by resistance that is either inherent or develops during treatment. Epothilones, which also stabilize microtubules but by a different mechanism, are in clinical development primarily to overcome taxane or multidrug resistance, based on potent preclinical antitumor activity against resistant tumor lines. Ixabepilone is the best-studied epothilone clinically and is active in patients with metastatic breast cancer that has been pretreated with, or had established resistance to, taxanes and/or anthracyclines. In a phase III trial in patients with anthracycline-pretreated or -resistant and taxane resistant locally advanced or metastatic breast cancer, adding ixabepilone to capecitabine significantly improved progression-free survival and the overall response rate compared with capecitabine alone. The primary toxicities associated with ixabepilone treatment are neuropathy and neutropenia, but both are generally manageable. Other epothilones currently in clinical studies are KOS-862, patupilone, ZK-EPO, BMS-310705, and KOS-1584, which have all shown activity in patients with pretreated or resistant metastatic breast cancer. PMID- 18410797 TI - Emerging agents in the treatment of anthracycline- and taxane-refractory metastatic breast cancer. AB - Despite the recent trend toward treatment of early stage breast cancer aggressively with anthracyclines and taxanes, nearly half of those women will have metastatic recurrence. Moreover, because of the increasing prior exposure to these drugs, far more women facing first-line therapy for recurrent disease will now have developed anthracycline- and taxane-refractory metastatic breast cancer (ATRMBC), presenting a major therapeutic challenge. A number of established drugs are showing promise in this setting: capecitabine alone or combined with lapatinib; gemcitabine; vinorelbine; and oxaliplatin. At the same time, a variety of new drugs are emerging for potential use in ATRMBC. Among the drugs in clinical development that have shown promising activity include novel classes of compounds (camptothecins and epothilones), newer members of established classes (pemetrexed and vinflunine), and agents with novel mechanisms of action (the mitosis inhibitor E7389 and the ascidian-derived anticancer compound trabectedin). Several molecularly targeted agents are also being evaluated in ATRMBC, including interleukin-2 receptor-binding denileukin diftitox, and 17 (allylamino)-17-demethoxygeldanamycin (17-AAG), which inhibits the protein chaperone heat shock protein 90. PMID- 18410798 TI - Cutaneous ultraviolet exposure and its relationship to the development of skin cancer. AB - Skin cancer is becoming an increasingly important public health problem. Multiple studies have now demonstrated a relationship between ultraviolet exposure and increased risk of developing skin cancer. However, the specifics of that association are somewhat different for malignant melanoma, basal cell carcinoma, and squamous cell carcinoma. A better understanding of the mechanisms that allow cutaneous ultraviolet radiation to induce neoplasia will result in the development of better future sun-protection agents and strategies. PMID- 18410799 TI - Rationale for sunscreen development. AB - The risks associated with cumulative or overexposure to ultraviolet (UV) radiation are now well identified. They involve both UVB (290-320 nm) and UVA (320-400 nm) components. UVB radiation is still considered to be the major factor responsible for most harmful effects of solar exposure. But the detrimental role of UVA has been increasingly documented. As a result, the development of a new generation of sunscreens that provide effective protection throughout the whole UV radiation spectrum has become a prime concern and a major issue. These full spectrum protections must be carefully designed by selecting UV filters with highly reliable stability to ensure optimal safety and efficiency during the entire period of sun exposure. For this purpose, extensive research on the photochemical behavior of various classes of UV filters has been carried out. This has led to the discovery of safe systems that prevent sun-induced degradation of potent UVA filters and to the development of new UVA filters with a high photostability. PMID- 18410800 TI - Skin DNA photodamage and its biological consequences. AB - It is well established that ultraviolet (UV) radiation from sunlight damages skin cells' DNA. Wavelengths in the UVB range are absorbed by DNA and can induce mutagenic lesions such as pyrimidine dimers. On the other hand, genotoxic effects of solar UVA are mainly mediated by the activation of endogenous photosensitizers resulting in the generation of a local oxidative stress. Exogenous chemicals, such as drugs like psoralens or fluoroquinolones, sometimes amplify UV-induced harmful effects. DNA damage can lead to mutations and genetic instability. This is one of the reasons why sunlight overexposure increases the risk of skin cancer. But DNA photolesions can also be involved in other skin-specific responses to UV radiation: erythema, immunosuppression, and melanogenesis are examples reported in the literature. The aim of this short review is to summarize the general knowledge in the field of UV-induced DNA damage. Besides the biological consequences of DNA photolesions, this article also deals with technologies used for their detection and shows how helpful such approaches can be to assess photoprotection provided by sunscreens. PMID- 18410801 TI - Broad-spectrum sunscreens provide better protection from solar ultraviolet simulated radiation and natural sunlight-induced immunosuppression in human beings. AB - BACKGROUND: It is well established that ultraviolet (UV) radiation induces immunomodulatory effects that may be involved in skin cancer. Recent studies have shown that UVA (320-400 nm) and UVB (290-320 nm) radiation are immunosuppressive. As a result, sunscreens, which mainly absorb UVB, may be less effective in preventing UV radiation-induced immunosuppression than broad-spectrum products. OBJECTIVE: We sought to study the effects of UVA exposure on human delayed-type hypersensitivity (DTH) response and compare the efficacy of sunscreens having different levels of sun-protection factor (SPF) and UVA protection against both solar-simulated radiation and outdoor real-life sunlight exposure conditions. METHODS: DTH was assessed using a kit which includes 7 recall antigens that most of the participants encountered during childhood immunization. Evaluation of DTH test response was made 48 hours after test application before and after UV exposure with or without sunscreens. RESULTS: In unprotected participants, the response to DTH tests was significantly reduced irrespective of UV types of exposure (full-spectrum UVA, long UVA, solar-simulated radiation). A UVB sunscreen failed to protect from solar-simulated radiation-induced immunosuppression. In contrast, a broad-spectrum sunscreen with the same SPF but providing a high protection in the UVA range significantly reduced local UV induced immunosuppression and prevented the distant effects. In the outdoor study, as compared with DTH responses obtained before sun exposure, no alteration of immune response was detected when the skin was protected by a broad-spectrum sunscreen having a high protection level in the UVA (SPF 25, UVA protection factor 14). Conversely a broad-spectrum sunscreen with lower protection against UVA (SPF 25, UVA protection factor 6) failed to prevent UV-impaired response. LIMITATIONS: These results have been obtained after repeated exposure. Additional experiments obtained under acute exposure are in progress. CONCLUSION: These findings clearly demonstrated the role of UVA in the induction of photoimmunosuppression together with the need for sunscreen products providing efficient photoprotection throughout the entire UV spectrum. PMID- 18410802 TI - An organotypic model of skin to study photodamage and photoprotection in vitro. AB - Acute or repetitive sun exposures are known to elicit cutaneous damages such as sunburn but also long-term effects such as photoaging or cancers. Determination of early biological events occurring after ultraviolet (UV) exposure is essential for photoprotection. Using skin reconstructed in vitro containing both a dermal equivalent and a fully differentiated epidermis, the effects of UV light (UVB and UVA) were investigated. UVB-induced damage was essentially epidermal, with the typical sunburn cells and DNA lesions, whereas UVA radiation-induced damage was mostly located within the dermal compartment. The model and end points used for UVB- and UVA-induced damages appeared to be very useful for the in vitro evaluation of sunscreens after topical application, in particular to investigate its protective effects against the effects of UVR, and allowed us to distinguish the efficiency of absorbers depending on their absorption spectrum. PMID- 18410803 TI - The benefit of daily photoprotection. AB - BACKGROUND: It is now recognized that both ultraviolet (UV)-A and UVB wavelengths participate in the generation of photodamaged human skin during sun exposure. During usual daily activities, an appropriate protection against solar UV exposure should prevent clinical, cellular, and molecular changes potentially leading to photoaging. OBJECTIVE: This study was designed to evaluate in human beings the protection afforded by a day cream containing a photostable combination of UVB and UVA filters and thus protect against the UV-induced skin alterations. RESULTS: In solar-simulated radiation exposed and unprotected skin sites we observed melanization. The epidermis revealed a significant increase in stratum corneum and stratum granulosum thickness. In the dermis, an enhanced expression of tenascin and a reduced expression of type I procollagen were evidenced just below the dermoepidermal junction. Although no change in elastic fibers in exposed buttock skin was seen, a slightly increased deposit of lysozyme and alpha-1 antitrypsin on elastin fibers was observed using immunofluorescence techniques. A day cream with photoprotection properties was shown to prevent all of the above-described alterations. LIMITATIONS: This study was performed on a limited number of patients (n = 12) with specific characteristics (20-35 years old and skin type II and III). Two dermal alterations were evaluated by visual assessment and not by computer-assisted image analysis quantification. CONCLUSION: Our in vivo results demonstrate the benefits of daily photoprotection using a day cream containing appropriate broad-spectrum sunscreens, which prevent solar UV-induced skin damages. PMID- 18410804 TI - Positive outcomes from an immediate and ongoing intervention for child witnesses of intimate partner violence. AB - OBJECTIVE: Children who witness intimate partner violence (IPV) experience many psychological and social problems similar to those of the victimized parent. Intervention programs for children who witness IPV have not been adequately evaluated. Two of the most important target areas in treating the children are to improve understanding that (1) the violence is not their fault and that (2) development of a safety plan is valuable in case of future violence. We evaluated a unique immediate and ongoing intervention program for children who witness adult IPV. METHODS: This was a retrospective review of progress report results both pre- and postintervention of an ongoing IPV intervention program. POPULATION AND INTERVENTION: The study was completed in a largely Hispanic city of 500,000. The intervention program included a number of unique aspects including immediate intervention at the time of police calls for adult IPV, children's art therapy and sand tray therapy, and a unique coloring book to establish a child safety plan in the event of recurrent IPV. Progress report forms included 16 qualitative questions that evaluated the child's understanding of various important concepts pre- and postintervention. INCLUSION: All children who entered the program in the last 3 years and had completed data sets for all 16 questions were included. Responses to questions pre- vs postintervention were compared using Wilcoxon signed ranks test. RESULTS: Fifty-eight children had complete data sets pre- and postintervention. Mean age was 8.5 +/- 3.5 years (range, 3-17), and 49% were male. Mean length of therapy was 7.4 +/- 5.2 months (range, 1-31), with a mean number of sessions of 9.7 +/- 11.7 (range, 1-59). For 15 of the 16 evaluation questions, a statistically significant improvement in postintervention evaluations compared with preintervention evaluations was found (P < .01). This included a significant improvement in the percentage of children who were aware that violence was not their fault (59% preintervention vs 84% postintervention; difference, 25%; 95% confidence interval, 9-41) and a significant improvement in the percentage of children who knew and understood a safety plan in case of recurrent episodes of violence exposure (32% preintervention vs 93% postintervention; difference, 61%; 95% confidence interval, 47-75). CONCLUSIONS: Among children who were exposed to adult IPV and subsequently underwent immediate and ongoing treatment, there was a significant improvement in the percentage of children who were aware that violence was not their fault and in the percentage of children who were aware of safety planning. PMID- 18410805 TI - Heart rate variability measures as predictors of in-hospital mortality in ED patients with sepsis. AB - OBJECTIVE: To determine the predictive capability of heart rate variability (HRV) measures of patients with sepsis in the ED for in-hospital death. METHODS: This was a prospective, observational study. A consecutive cohort of patients visiting the ED of a university teaching hospital who met the criteria of sepsis over a 6 month period were enrolled in this study. General demographics, vital signs, laboratory data, and Mortality in Emergency Department Sepsis score were obtained in the ED; the in-patient medical record was reviewed; and a series of continuous 10-minute electrocardiographic signals were recorded for off-line HRV analysis to assess the in-hospital mortality of the patients. RESULTS: One hundred thirty-two patients aged 27 to 86 years who met the inclusion criteria were enrolled. According to the in-hospital outcome, the patients were categorized into 2 groups: nonsurvivors (n = 10) and survivors (n = 122). The baseline HRV measures, including SDNN, TP, VLFP, LFP, and LFP/HFP ratio, of nonsurvivors were significantly lower, whereas the nHFP was significantly higher, than those of survivors. Multiple logistic regression model identified SDNN and nHFP as the significant independent variables in the prediction of in-hospital mortality for ED patients with sepsis. The receiver operating characteristic area for SDNN and nHFP in predicting the risk of death was 0.700 and 0.739, respectively. CONCLUSIONS: Heart rate variability measures, especially the SDNN and nHFP, may be used as valuable predictors of in-hospital mortality in patients with sepsis attending the ED. PMID- 18410806 TI - The efficacy of low-dose antivenom therapy on morbidity and mortality in snakebite cases. AB - Similar to the cases seen around the world, snakebite causes mortality and morbidity in Turkey. The venom of different types of snake in the region of Cukurova causes serious systemic and local tissue damage. METHODS: We performed this prospective study on 45 patients who complained of snakebite. We grouped the patients according to their clinical presentations to facilitate treatment and follow-up period procedures. RESULTS: Whereas the patients with grade 1 envenomation did not receive any antivenom, grades 2 and 3 patients received 2.70 +/- 0.77 and 4.88 +/- 1.65 vials of antivenom, respectively. One of our patients had to undergo finger amputation but there were no deaths. Allergic reactions developed after antivenom therapy in 8 patients (17.8%). CONCLUSIONS: When we considered the cost and complications of the antivenom treatment, it was seen that low-dose antivenom treatment effectively treated the patients with venomous snakebite injuries in our region. PMID- 18410807 TI - Admission ST-segment elevation in lead aVR as the factor improving complex risk stratification in acute coronary syndromes. AB - This study aimed to analyze the prognostic value of the presence of ST elevation in lead aVR [aVR(+)] in initial standard electrocardiogram (ECG) performed on admission in combination with clinical variables and Thrombolysis in Myocardial Infarction (TIMI) risk score for unstable angina/non-ST-elevation myocardial infarction (UA/NSTEMI). In 205 consecutive patients with UA/NSTEMI, we retrospectively evaluated admission ECG for aVR(+) of more than 0.5 mm. With the use of multivariate analysis, admission aVR(+) was found to be a strong and independent predictor of 30-day mortality. Mortality also increased with the severity of aVR(+): 2.2%, 10.8%, 13.8%, 22.2%, 50% (P value for trend <.0001). In prespecified low-risk groups by clinical factors, those with aVR(+) had higher death rates than those without aVR(+): 16.1% vs 2.2% (P = .04), 13.9% vs 1.1% (P = .001), 12.4% vs 1.1% (P = .002), 9.6% vs 1.2% (P = .02), and 6.7% vs 0% (P = .05) for patients with negative troponin, heart rate of 110 beats per minute or less, systolic blood pressure greater than 90 mm Hg, Killip I class on admission, and age 70 years or younger, respectively. Patients with aVR(+) compared to patients without aVR(+) had higher death rates in the low- and intermediate-risk groups by TIMI risk score. Our findings suggest that aVR(+) has significant prognostic value in patients with UA/NSTEMI and may provide an additional prognostic value to the conventional cardiovascular risk factor, particularly in patients in the low-risk and intermediate-risk groups. PMID- 18410808 TI - Neurologic and neuropsychological symptoms during the first year after an electric shock: results of a prospective multicenter study. AB - OBJECTIVES: To assess the prevalence of neurologic and neuropsychological symptoms in the short-term and 1 year after an electric shock and to explore whether any of these were associated with risk factors. METHODS: Patients presenting to one of 21 EDs between October 2000 and November 2004 were eligible to be enrolled in a prospective observational study after an electric shock if they had risk factors for late arrhythmias. Telephone follow-up was done to evaluate the appearance of symptoms. RESULTS: A total of 30 (26%) of 114 patients complained of neurologic or neuropsychological symptoms at a median of 52 days post-electric shock. At 1 year, 24 (28%) of 86 patients complained of neurologic or neuropsychological symptoms. None of the risk factors evaluated were associated with the symptoms. CONCLUSION: The prevalence of the symptoms we observed should alarm all emergency physicians that the effect of electricity can cause late neurologic and neuropsychological manifestations. PMID- 18410809 TI - The analysis of risk factors of impacting mortality rate in severe multiple trauma patients with posttraumatic acute respiratory distress syndrome. AB - OBJECTIVE: We hypothesize that not all of the traditional risk factors of impacting mortality rate in commonly traumatic populations with posttraumatic acute respiratory distress syndrome (ARDS) are independently associated with those patient populations identified with severe multiple trauma. Rather, we postulate that there may exist significantly different impacting degrees of specific risk factors in stratified patients (surviving beyond 24 and 96 hours)- more severe multiple trauma with higher injury score and long-term mechanical ventilation as well. METHODS: This is a retrospective cohort study regarding trauma as a single cause for emergency intensive care unit admission. Twenty-two items of potential risk factors of impacting mortality rate were calculated by univariate and multivariate logistic analyses to find distinctive items in these severe multiple trauma patients. RESULTS: The unadjusted odds ratio and 95% confidence intervals of mortality rate were found to be associated with 6 (out of 22) risk factors, namely, (1) Acute Physiology and Chronic Health Evaluation II score, (2) duration of trauma factor, (3) aspiration of gastric contents, (4) sepsis, (5) pulmonary contusion, and (6) duration of mechanical ventilation. Significant results also appeared in stratified patients. CONCLUSIONS: Impact of pulmonary contusion and Acute Physiology and Chronic Health Evaluation II score contributing to prediction of mortality may exist in the early phase after trauma. Sepsis is still a vital risk factor referring to systemic inflammatory response syndrome, infection, secondary multiple organ dysfunction, etc. Discharging trauma factors as early as possible becomes the critical therapeutic measure. Aspiration of gastric contents in emergency intensive care unit admission could lead to incremental mortality rate due to aspiration pneumonia. Long-standing mechanical ventilation should be constrained because it is likely to cause severe refractory complications. PMID- 18410810 TI - The effect of different relieving methods on the outcome of out-of-hospital cardiac arrest patients with nontraumatic hemopericardium in the ED. AB - AIMS: This study aimed to assess the impact of different methods of draining nontraumatic hemopericardium on outcome from patients with out-of-hospital cardiac arrest (OHCA), identify independent predictors of return of spontaneous circulation (ROSC), and examine the ineffective rate of decompression based on subxiphoid pericardiotomy (SP) and percutaneous pericardial catheter drainage (PCD). METHODS: Adult patients with OHCA who presented to the ED between May 1, 2000, and October 30, 2006, with moderate to massive nontraumatic hemopericardium were recruited and stratified into 4 groups according to the relieving methods of hemopericardium. Charts were reviewed for various demographic data, resuscitation records, management, and outcome. Patient outcome was recorded as survival to hospital discharge and ROSC, as primary end points. Effective decompression was recorded as a secondary end point. We compared the outcome between the groups. RESULTS: A total of 1491 OHCA resuscitation records were prospective collected. There were 23 OHCA patients with moderate to massive nontraumatic hemopericardium. The overall ROSC rate was 39.1% (9/23). There was a clear difference in the ROSC rate between 4 groups (P < .05). The overall rate of survival to hospital discharge was 4.3% (1/23). There was no significant difference in the rate of survival to hospital discharge between the groups. Relieving methods was an independent predictor of ROSC in OHCA patients with nontraumatic hemopericardium (odds ratio, 0.17; 95% confidence interval, 0.4 0.70). There was a significant statistical difference in adequate relief of hemopericardium based on SP and PCD (P < .01). CONCLUSION: The early effective decompression method is associated with an increased rate of ROSC for OHCA patients with nontraumatic hemopericardium. Subxiphoid pericardiotomy has a better effective decompression of hemopericardium than PCD. PMID- 18410811 TI - Hair apposition technique for scalp laceration repair: a randomized controlled trial comparing physicians and nurses (HAT 2 study). AB - OBJECTIVES: The hair apposition technique (HAT) is a new method of closing scalp lacerations in which hairs on either side of the wound are twisted together and secured with a tissue adhesive. We aimed to compare the effectiveness, complications, and benefits of HAT performed by nurses or doctors in a randomized, prospective trial. METHODS: We conducted the study in the ED from November 2002 to February 2005. Subjects were randomized to receive HAT either by doctors or nurses. All wounds were evaluated 7 days later. The outcomes wound infection, wound healing, bleeding, and overall complications were measured, setting +/-5% in the differences of the outcomes between the doctors and nurses as equivalence. RESULTS: There were 88 and 76 patients in the doctor and nurse groups, respectively. There were no significant differences in all short-term outcomes between the doctors and nurses except for length of the procedure. The doctors had a shorter mean duration of procedure than the nurses (9.0 +/- 5.6 vs 12.8 +/- 7.5 minutes, P = .001). CONCLUSION: The HAT can be safely performed by trained nurses with equivalent outcomes as doctors. PMID- 18410812 TI - Prognostic value of the Thrombolysis in Myocardial Infarction risk score in a unselected population with chest pain. Construction of a new predictive model. AB - INTRODUCTION: The Thrombolysis in Myocardial Infarction (TIMI) risk score (TRS) has proven to be a useful and simple tool for risk stratification of patients with chest pain in intermediate- and high-risk populations. There is little information on its applicability in daily clinical routine with unselected populations. AIMS: The aims of the study were to prospectively analyze the predictive value of the TRS in a heterogeneous population admitted for chest pain and to construct where possible a new modified model with a greater prognostic capacity. POPULATION AND METHODS: Seven hundred eleven consecutive patients were admitted over a 1-year period to the cardiology unit for chest pain without ST segment elevation. Thrombolysis in Myocardial Infarction risk score variables, relevant medical history variables, in-hospital examination results, and therapy information were collected. Cardiac events at 1 and 6 months were recorded. RESULTS: Seventy-one (9.8%) patients had a compound event (myocardial infarction/revascularization/cardiac death) at 6 months. On multivariate analysis, the variables associated with cardiac events were left ventricular ejection fraction (EF) of <35% (hazard ratio [HR] = 2.9, P = .002), diabetes (HR = 1.8, P = .02), and TRS (HR = 1.3, P = .007). Events at 6 months were 2.3% for a TRS of 0/1, 4.2% for 2, 10.2% for 3, 11.0% for 4, and 18.7% for a score of more than 5. A new modified scale was constructed to include EF and diabetes as independent variables, and this yielded an increase of 44% in the combined event at 6 months per score unit increase (HR = 1.44, P = .001). The modified scale showed a greater predictive capacity than the original model. CONCLUSIONS: The TRS is an important short- and long-term prognostic predictor when applied to an unselected population consulting for chest pain. The inclusion of diabetes and EF as variables in the model increases predictive capacity at no expense to simplicity. PMID- 18410813 TI - Predicting the development of anemia by measuring the diameter of the inferior vena cava of patients with spinal cord injury. AB - PURPOSE AND METHODS: We retrospectively investigated whether measurement of the diameter of the inferior vena cava (IVC) is useful in predicting the development of anemia in 12 patients with spinal cord injury. Measurement of the IVC was performed in conjunction with computed tomography scan upon arrival. The subjects were divided into two groups: those with a minimum (Min) hemoglobin (Hb) of less than 10 g/dL, and those with a Min Hb greater than 10 g/dL during hospitalization. RESULTS: The average diameter of IVC in the Hb<10 g/dL group was smaller that that in the Hb>10g/dL group. Blood pressure was not significantly different between the two groups. The difference between the Min and initial Hb value and incidence of blood transfusion in the Hb<10g/dL group was significantly greater than that in the Hb>10g/dL group. CONCLUSION: To predict the development of anemia, measurement of the IVC of patients with spinal cord injury may therefore be useful. PMID- 18410814 TI - Derivation of a clinical prediction rule for evaluating patients with abdominal pain and diarrhea. AB - OBJECTIVE: The objective of the study was to develop a simple prediction rule to reliably identify abdominal pain patients with diarrhea who may require surgical intervention. METHODS: We performed a secondary analysis of a prospective cohort study of adults with acute nontraumatic abdominal pain and diarrhea in an urban emergency department (ED). Structured data collection included 109 historical and 28 physical examination items, laboratory and radiographic results, and final diagnosis. The main outcome was operative intervention. RESULTS: One thousand patients were enrolled; 174 patients with diarrhea were included in this analysis. Patients had a mean age of 39 +/- 16 years and were likely to be female (64%) and black (60%). Fifteen (9%) patients received a surgical intervention from the ED. Clinical variables associated with the need for surgical intervention using univariate analysis were age older than 40 years, constant pain, and peritonitis on examination. Using recursive partitioning multivariate analysis, the derived prediction rule included 2 variables: age older than 40 years and constant pain. This rule had a sensitivity of 1.0 (95% confidence interval, 0.78-1.0) and specificity of 0.23 (95% confidence interval, 0.16-0.30). CONCLUSION: Patients older than 40 years with constant abdominal pain and diarrhea are likely to have a surgical cause of their symptoms. PMID- 18410815 TI - Does lack of social support lead to more ED visits for older adults? AB - OBJECTIVE: The goals of this study were to (1) determine whether level of social support and living situation predicted emergency department (ED) use among older adults and (2) identify correlates of ED visits according to whether the patient was admitted to the hospital. METHODS: Secondary analysis of a longitudinal, prospective study was conducted. RESULTS: In adjusted analyses, subjects who lived alone were 60% more likely to visit the ED than those who lived solely with their spouse. Neither type nor level of social support as measured by the Duke Social Support Index predicted ED use. Indicators of poor physical health (prior hospitalization, poorer self-rated health, and functional disability) were predictors of ED visits that resulted in hospitalization; however, these were not significantly associated with outpatient ED visits. DISCUSSION: Older adults who live alone are more likely to visit the ED. Additional study is needed to understand the determinants of outpatient ED visits. PMID- 18410816 TI - Are racial disparities in ED analgesia improving? Evidence from a national database. AB - OBJECTIVE: Ethnic disparities in emergency department (ED) analgesic use have been noted previously; the purpose of this study was to determine if current ED practice has been altered subsequent to the widespread recognition of these inequalities. METHODS: Using data from the National Hospital Ambulatory Care Survey, we analyzed ED analgesic treatment with respect to race for the complaints of headache (HA) and long bone fractures (LBF) for 1995 to 1999 (period A) and compared that with data for 2000 to 2003 (period B). We compared the use of "any analgesics" and "opioids" among blacks, whites, and Hispanics. RESULTS: For both HA and LBF, improvements over time were noted in all 3 ethnic subgroups. A statistically significant increase was documented in the treatment of HA among Hispanics using any analgesia (71.3% vs 80.8%, P = .011). Although individual differences between the 2 periods in the LBF cohort were not statistically significant for any of the 3 ethnic groups, there was an overall improvement noted when combining all ethnicities: among all patients treated for LBF, 66% received some analgesic and 42.8% received opioid analgesia during period A vs 75% and 51%, respectively, during period B (all P < .001). Furthermore, in the latest study period, there were no differences in the frequency of analgesic administration for LBF with respect to race (blacks, 43%; whites, 48%; Hispanics, 43.8%; P > .1). CONCLUSION: There is evidence that previously described racial inequalities in analgesic use have decreased over time. PMID- 18410817 TI - Trauma systems and timing of patient transfer: are we improving? AB - INTRODUCTION: The regionalization of trauma services is based on the premise that injured persons presenting to nontertiary facilities will be stabilized and rapidly transported to a more definitive center. Although trauma systems seem to improve outcomes for urban patients, this same benefit has not been shown for rural patients. There are many factors associated with the decision to transfer injured patients to a regional trauma center, including referral hospital and patient age, for example. The purpose of this study is to examine factors that influence the timing of transfer of trauma patients and specifically to determine if establishing specific trauma systems has led to any changes in transfer timing over time. METHODS: The trauma registry at the University of Wisconsin was queried for all patients admitted between July 1, 1999, and June 30, 2005. Patients were included in this study if they had been transferred to the university hospital after evaluation at an outside hospital. The registry variables that were abstracted were age, referring hospital, emergency department (ED) time at referring hospital, injury severity score (ISS), the presence of a head injury, performance of a head computed tomography (CT), mode of transport, and the date of ED evaluation. RESULTS: There were 1656 patients with ISS higher than 9 transferred during the period. The mean ED time was 153 +/- 82 minutes. Emergency department time was significantly shorter for those with ISS higher than 25 and for those transported by helicopter. Four hundred ninety-two (30%) patients had a head CT performed at the outside hospital, of which 221 (44%) were repeated at the trauma center. The mean ED time for those in whom a CT was performed was significantly longer than those without CT (179 +/- 81 vs 142 +/- 84 minutes). The ED times were slightly longer for level III hospitals (158 +/- 82 minutes) than for level IV hospitals (137 +/- 74 minutes). Emergency department times were longer for older patients. The times in the ED showed an upward, but not statistically significant, trend. After controlling for all other variables, ED times were not significantly different over the period studied. CONCLUSION: Development of a statewide trauma system and outreach education has not significantly affected transfer times from nontrauma centers in our system. Outreach educational efforts should focus on systematic trauma evaluation, prompt transfer, and limitation of nontherapeutic testing. PMID- 18410818 TI - Inadequate provision of postintubation anxiolysis and analgesia in the ED. AB - INTRODUCTION: Patients intubated in the emergency department (ED) often have extended ED stays. We hypothesize that ED intubated patients receive inadequate postintubation anxiolysis and analgesia after rapid sequence induction (RSI). METHODS: This was a retrospective cohort study of every adult intubated in a tertiary-care ED (July 2003-June 2004). Patients were included if they underwent RSI, remained in the ED for more than 30 minutes post intubation, and survived to admission. Presuming a mean patient weight of 70 kg, we defined adequacy of anxiolysis and analgesia on the provision postintubation of weight-based doses of lorazepam (0.77 mg/h) or midazolam (4.2 mg/h) and fentanyl (35 microg/h), referenced from pharmaceutical texts. Demographic data, time in ED, and dosage of each medication given were abstracted. The proportion, with 95% confidence intervals (CIs), of patients receiving inadequate anxiolysis and analgesia were computed. RESULTS: One hundred seventeen patients met the inclusion criteria. Mean time in the ED was 4.2 hours (SD +/- 3.1 hours). Thirty-nine patients received no anxiolytic (33%, CI 25%-43%), and 62 received no analgesic (53%, CI 44%-62%). Twenty-three patients received neither anxiolytic nor analgesic (20%, CI 13%-28%). Of 70 patients given postintubation vecuronium, 67 received either no or inadequate anxiolysis or analgesia (96%, CI 87%-99%). Overall, 87 of 117 patients received no or inadequate anxiolysis (74%, CI 65%-82%); and 88 of 117 received no or inadequate analgesia (75%, CI 66%-83%). CONCLUSION: Patients undergoing RSI in the ED frequently receive inadequate postintubation anxiolysis and analgesia. PMID- 18410820 TI - Principles of war surgery: current concepts and future perspectives. AB - Medical support is an important part of military operations. The aim of war surgery is to achieve the return of the greatest number of injured to combat and the preservation of life, limb, and eyesight. War surgery is different from current traumatology because of many reasons. Because hemorrhage is the most common cause of death in military trauma, airway preservation and effective control of bleeding represent the highest priorities in war injuries. Wound excision (the so-called debridement) is a significant part in the management of war injuries. It involves excision of all foreign objects and contaminants and dead/nonviable tissue that--if not removed--would become a medium for infection. Broad-spectrum antibiotics should be administered and tetanus prophylaxis measures should be taken, as indicated. Delayed wound closure (usually after 4-5 days) is the standard procedure after wound excision. Recently, changes in the dogma of war necessitated significant changes in the organization schema of military services supporting modern military operations. The concept of highly mobile, easily deployed, forward surgical facilities is the most important change in the philosophy of modern war injury. Military surgeons are now facing new challenges; appropriate education is required to achieve success in their mission. PMID- 18410819 TI - Assessment of severity measures for acute asthma outcomes: a first step in developing an asthma clinical prediction rule. AB - OBJECTIVE: As a first step in the development of an asthma prediction rule, our primary objective was to assess the association of 8 candidate predictor variables with 2 clinically relevant asthma outcomes. METHODS: Among a cohort of 125 adults hospitalized with an asthma exacerbation, we examined models to identify clinical variables associated with length of stay (LOS) and clinically significant asthma exacerbations within 3 months after hospitalization (3-month exacerbation). Eight candidate predictor variables were chosen, including age, sex, race, pulsus paradoxus, prior endotracheal intubation for asthma, hospitalization within 5 years for asthma, and 2 chronic asthma severity scores. RESULTS: We found independent associations between LOS and pulsus paradoxus (P = .005), prior intubation (P = .03), sex (P = .03), and prior hospitalization (P = .019). Among men, 52% had a 3-month exacerbation in comparison with 25% of women; and in multivariable analysis, male sex was independently associated with 3-month exacerbation (adjusted odds ratio = 5.1; 95% confidence interval = 1.37-18.9; P = .015). Participants with 3-month exacerbation had higher Johns Hopkins Allergy and Asthma Composite (JHAAC) chronic severity scores (median = 77; interquartile range = 57-91) than those who did not (median = 54; interquartile range = 35-69; P < .001) (for 40-unit increase, adjusted OR for 3-month exacerbation = 1.54; 95% confidence interval = 1.16-2.03; P = .003). In multivariable analysis, male sex and the JHAAC severity score were independently associated with 3-month exacerbation. CONCLUSIONS: Elevated pulsus paradoxus, prior intubation for asthma, and 5-year asthma hospitalization are independently associated with LOS. Race, 5-year asthma hospitalization, and JHAAC score predict 3-month asthma exacerbation. These variables warrant consideration for use in the development of an asthma prediction rule. PMID- 18410821 TI - The cyanoacrylate topical skin adhesives. AB - Each year there are over 7 million lacerations requiring wound closure in the emergency department. Traditionally, most lacerations have been closed with sutures. Topical cyanoacrylate skin adhesives offer many advantages over traditional wound closure devices. Recently, the Food and Drug Administration (FDA) has reclassified the topical skin adhesives. As a result, new topical skin adhesives are expected to enter the market in the near future. This article will review the structure and function of cyanoacrylates as well as their advantages, indications, and usage. PMID- 18410822 TI - The pediatric electrocardiogram part III: Congenital heart disease and other cardiac syndromes. AB - Approximately 1% of newborns are affected by congenital heart disease (CHD), and although many lesions of CHD have trivial hemodynamic and clinical implications, some clinically significant lesions are asymptomatic in the immediate newborn period and may present after discharge from the well baby nursery. Because of this, CHD should be considered in the differential diagnosis of any ill-appearing newborn, regardless of the presence of cyanosis. In addition, the number of children, adolescents, and adults with surgically repaired or palliated CHD continues to grow within the United States and other developed countries. It is in this population that arrhythmias are particularly prone to develop, and knowledge of the common arrhythmias associated with CHD is mandatory for the acute care provider. PMID- 18410823 TI - Conflict of interest in research. PMID- 18410825 TI - Acute myeloid leukemia presenting as one-and-a-half syndrome. AB - One-and-a-half syndrome is a clinical disorder featuring extraocular movements characterized by horizontal conjugate gaze palsy with internuclear ophthalmoplegia. It usually results from a unilateral lesion of the midbrain, and the most common cause of this syndrome in young women is multiple sclerosis. We report the case of a 38-year-old woman diagnosed as having acute myeloblastic leukemia presenting with characteristic neurologic and imaging features of one and-a-half syndrome. Hyperleukocytosis, cancer procoagulants, tissue factor expression, and the increased proteolysis of coagulation factors by leukemic cells may all contribute to the propensity for thrombotic vascular occlusion. The optimal treatment of acute brain infarction in acute leukemia patients with hyperleukocytosis remains unclear. However, this patient illustrates that leukapheresis alone can provide rapid and effective relief of visual symptoms without neurologic sequela. To achieve better outcomes and survival, clinicians must maintain a heightened awareness of this distinctly unusual manifestation. PMID- 18410826 TI - Not all acute lower back pain is benign--paravertebral abscess and colonic cancer. AB - Low back pain with sciatica is one of the most common complaints of patients presenting to the ED, and it is usually managed on an outpatient basis. However, acute lower back pain not always derives from a benign cause. We report here the case of a 63-year-old diabetic man who presented to the ED complaining of acute low back pain with sciatica and fever. The cause was a large paravertebral abscess by Streptococcus milleri, and this was the first presenting sign of an unknown underlying colonic cancer. PMID- 18410827 TI - An under-recognized complication of treatment of acute severe asthma. AB - A 39-year-old man presented to the emergency department (ED) in severe respiratory distress. He had a prior diagnosis of brittle asthma and had been admitted on several occasions but never previously ventilated. Therapy given in the first 3 hours of arrival included nebulized salbutamol (5 mg, x5), ipratropium bromide (0.5 mg), intravenous hydrocortisone (200 mg), and magnesium sulfate (2 g). His arterial blood gases continued to deteriorate. He was then given an intravenous bolus of salbutamol (250 microg) and heliox via facemask. His worsening status necessitated invasive ventilation. His hypercapnia and resultant respiratory acidosis improved rapidly, but there was a concurrent accumulation of lactic acid resulting in acidemia. This patient had lactic acidosis as a direct effect of administration of salbutamol. The development of hazardous salbutamol-induced toxicity in acute severe asthma is discussed. PMID- 18410828 TI - Spontaneous septic diskitis: a common complaint with a serious pathologic cause that should not been overlooked. AB - Many people who have low back pain are likely to be out of work and taking medication and probably make demands on both primary and secondary health care as well as the private sector. Septic diskitis is a rare cause of back pain, accounting for less than 0.01% of cases in the primary care setting (Carragee EJ. Pyogenic vertebral osteomyelitis. J Bone Joint Surg Am. 1997;79:874-80). It has been associated with a high morbidity and mortality in the past. Although it is an unusual complication in a postoperative patient, it is even more rare in a nonoperative patient. It accounts for 2% of all osteomyelitis (Maiuri F, Iaconetta G, Gallicchio B, Manto A, Briganti F. Spondylodiscitis. Clinical and magnetic resonance diagnosis. Spine. 1997;22:1741-6) and may occur spontaneously, after surgery (especially after epidural anesthesia), in the immunosuppressed, and systemic infections. It has the potential to be an orthopedic pitfall in the emergency department because it can be presented with nonspecific symptoms, nonspecific laboratory abnormalities, and nondiagnostic radiographic studies. We present a case of low back pain in a previously healthy individual as a result of spontaneous septic diskitis, which had been initially misdiagnosed in the private sector as herniation of an intervertebral disk. PMID- 18410829 TI - Adolescent pneumopericardium and pneumomediastinum after motor vehicle crash and ejection. AB - A 15 year old male was an unrestrained passenger in a high speed motor vehicle crash followed by ejection. The patient was noted to have evidence of bilateral pneumothorax upon arrival in the Emergency Department. Bilateral chest tubes were placed under sterile conditions; however, the left pneumothorax remained, and a second left chest tube was placed. Repeat chest radiographs revealed extensive subcutaneous emphysema, pneumomediastinum, and pneumopericardium. Needle aspiration of the pericardium returned significant quantities of air, an immediate improvement in blood pressures followed. An 18-gauge triple lumen catheter was placed into the pericardial space for additional withdrawal of air via syringe. Mechanisms have been proposed to explain the development of tension pneumopericardium after chest trauma. Early diagnosis is crucial, and may be found on initial chest radiographs. Computed tomography is also an effective method for evaluating the presence of air in the pericardial space and may assist in establishing the diagnosis. Tension pneumopericardium requires immediate recognition and decompression to prevent cardiac tamponade with a fatal circulation collapse, an entity that is as serious as the tamponade resulting from hemopericardium. Traumatic pneumopericardium is rare, but can be a complicated finding associated with high-speed blunt chest trauma. Patients with evidence of pneumopericardium should be closely monitored, particularly those supported by positive pressure ventilation. PMID- 18410830 TI - Delayed hypersensitivity reaction from black henna tattoo manifesting as severe facial swelling. AB - We report on a 14-year-old boy who was presented to the emergency department with an acute swelling of the face and scalp 3 days after using a new hair dye. The patient had applied a black henna tattoo 1 year earlier. Patch testing revealed an allergy to the potent skin sensitizer paraphenylenediamine, a common ingredient of hair dyes and also found in black henna tattoo. It is important for emergency physicians to be aware of the possibility of a delayed type-IV hypersensitivity reaction from black henna tattoos manifesting as an acute contact dermatitis. These patients may have gross facial swelling but should not be treated for angioedema. PMID- 18410831 TI - A rare incidental finding in a case of painless jaundice. AB - Portal vein thrombosis secondary to protein C deficiency is a rare finding. Diagnosing a portal vein thrombosis itself is difficult due to nonspecific symptoms such as nausea, vomiting, anorexia, and weight loss. Proving that a protein C deficiency is the cause of a portal vein thrombosis is even more difficult as an extensive and thorough workup is required to rule out malignancies, myeloproliferative disorders, and hypercoaguable states which can all lead to thromboses. Patients require anticoagulation to prevent two dangerous complications of portal vein thrombosis; portal hypertension leading to esophageal varices with massive hemetemesis and extension of thrombus from the portal vein into the mesenteric veins leading to intestinal ischemia and death. In this case report, we present a patient with the complaint of painless jaundice who was found to have an incidental finding of portal vein thrombosis secondary to protein C deficiency. The different etiologies of portal vein thrombosis, along with diagnosis and treatment options will be discussed and highlighted. PMID- 18410832 TI - Transarterial embolization in the management of life-threatening hemorrhage after maxillofacial trauma: a case report and review of literature. AB - There are many reasons for hypotension in trauma patients. Life-threatening hemorrhage associated with maxillofacial trauma is considered rare. Here, we present a 25-year-old patient with maxillofacial trauma complicated by life threatening hemorrhage after a traffic accident. At the emergency department, massive epistaxis was noted. Nasal packing and blood transfusion were performed, but vital signs of the patient were still unstable. Cerebral angiography revealed contrast extravasation from the left superficial temporal and internal maxillary arteries of the left external carotid artery. After transarterial embolization was performed, the hemorrhage immediately stopped. When common treatment such as nasal packing, correction of coagulopathy, reduction of fractures, and arterial ligation fails to control the hemorrhage, transarterial embolization can offer a safe alterative to surgical exploration. PMID- 18410833 TI - An avoidable abdominal surgery: pneumatosis coli. AB - We report on the case of a 90-year-old man who presented to the emergency department with constipation for 1 week and abdominal fullness for 2 days. Abdominal plain film radiography disclosed intramural air in the colon, which indicated pneumatosis coli (PC). Exploratory laparotomy was performed immediately under the impression of ischemic bowel disease. Through examination of the mesentery, the intestine and colon revealed no sign of perforation and ischemia. Surgery for PC is limited to patients with signs of perforation, peritonitis, intra-abdominal abscess, or bowel ischemia. Conservative treatment with oxygen supply, hyperbaric oxygen therapy, and antibiotics remain to be the mainstay for most patients with PC. PMID- 18410834 TI - Pelvic ectopic kidney with acute pyelonephritis: wolf in sheep's clothing. AB - The differential diagnosis of right lower quadrate pain at the emergency department is quite perplexing. We describe a 38-year-old woman presenting with characteristic clinical and laboratory features of ruptured appendicitis with severe sepsis. However, contrast-enhanced computed tomography scan of the abdomen established the diagnosis of pelvic ectopic kidney with acute pyelonephritis. Antibiotic treatment eventually achieved satisfactory resolution without compromise of renal function. It is should be addressed that, even with the advent of modern imaging modalities, there can be a diagnostic pitfall in general practice when managing right lower quadrate pain in patients with typical presentations of acute appendicitis without sonographic evidence, as illustrated in this case. In conclusion, early recognition using exquisite imaging studies with raised awareness in the clinical setting and prompt antibiotic treatment can avoid unnecessary intervention, preserve renal function, and prevent a life threatening catastrophe. PMID- 18410835 TI - Occult lacerations to the epiglottis and pharynx by glass fragments. AB - Injury to the epiglottis and pharynx is very rare in neck trauma overall. We experienced such a rare case in which a 43-year-old woman had lacerations to the epiglottis and pharynx. It was difficult to determine the exact site of the damage by observing the cut. We believed there was no damage to the lower respiratory tract because her vital signs were stable and her consciousness was clear. However, initial computed tomography revealed emphysema and showed that fragments from a glass vase had pierced the posterior pharyngeal wall. PMID- 18410836 TI - Acute hydrogen sulfide toxicity due to sewer gas exposure. AB - Hydrogen sulfide toxicity is a known risk for individuals working in the petroleum, sewer, maritime, and mining industries. Concern regarding exposure has led to the development of safety precautions and treatment guidelines. The US government imposes safety measures including self-contained breathing masks and exposure time limits to hydrogen sulfide gas. Current treatment methods, however, are not strongly supported by research. Acute exposure to hydrogen sulfide gas still poses a significant life threat. In this report, we discuss a case of a sewer worker exposed to deadly concentrations of hydrogen sulfide. Safety precautions and treatment options available to those exposed to high doses of hydrogen sulfide gas are explored. PMID- 18410837 TI - Proton pump inhibitor-induced rhabdomyolysis and hyponatremic delirium. AB - Many consider proton pump inhibitors (PPIs) to be devoid of adverse effects; however, PPIs can cause serious adverse effects. We report on the case of an emergency department patient with omeprazole-induced hyponatremic delirium and rhabdomyolysis. Proton pump inhibitors are initially overlooked as a cause of hyponatremia and myopathy in the emergency department, as with our case. This case is also unique because no publication has reported on both hyponatremia and severe myopathy after PPI therapy. The patient recovered completely after cessation of omeprazole and initiation of intravenous saline hydration and desmopressin infusion. PMID- 18410838 TI - Backward somersault as a cause of childhood stroke: a case report of isolated middle cerebral artery dissection in an adolescent boy. AB - Arterial dissection is a rare but serious cause of acute childhood stroke. Here, we present the case of a 15-year-old adolescent boy, an art school student, who suddenly experienced left hemi-paralysis after performing repeated backward somersaults, diagnosed as right-sided middle cerebral artery (MCA) hyperacute infarct due to isolated MCA dissection. Middle cerebral artery dissection in childhood is rare, as in our patient; however, pediatricians and emergency physicians should take MCA dissection into consideration in the differential diagnosis when hyperdense MCA sign is detected from unenhanced cranial computed tomography (CT) of childhood and/or adolescent stroke, with a special history such as backward somersault as in this case. Besides, unenhanced cranial CT and further imaging studies, including CT angiography and magnetic resonance imaging, provide a crucial role in early diagnosis to design proper treatment of acute childhood stroke in the ED. PMID- 18410839 TI - Unusual presentation of acute ovarian torsion in an adolescent. AB - Ovarian torsion occurs when an ovary twists about its vascular pedicle. The process causes abdominal pain and will result in ovarian tissue loss if not diagnosed and managed expeditiously. Acute ovarian torsion is an uncommon cause of abdominal pain in adolescents, and, as a result, it is often mistaken for other processes. We present a case of ovarian torsion in an adolescent female who presented with abdominal pain and marked abdominal distension, which was initially suggestive of a gravid abdomen. She was ultimately found to have a very large cystic teratoma of the right ovary with concomitant ovarian torsion. There are no reported cases of ovarian cystic teratoma with acute torsion presenting with an abdominal mass suggestive of a gravid abdomen. PMID- 18410840 TI - Posterior myocardial infarction: unique diagnosis to an elusive problem. AB - The clinical presentation of posterior myocardial infarction is not easy. The diagnosis is often missed due to lack of ST-segment elevation in standard 12-lead electrocardiogram. The diagnosis is made by seeing ST-segment elevation in the posterior leads V7, V8, and V9, which are typically placed in the left posterior axillary line, left midscapular line, and halfway between the mid scapular and left paraspinal line, respectively. The investigators describe a case of posterior myocardial infarction where additional posterior leads were placed in the left paraspinal line, right paraspinal line, and right midscapular line, displaying more prominent current of injury than seen with traditional posterior lead placement. This may lead to a more robust identification of posterior myocardial infarction that, in turn, may allow for adequate treatment and triage. PMID- 18410841 TI - Pseudo-subarachnoid hemorrhage. AB - The computed tomographic (CT) finding of pseudo-subarachnoid hemorrhage (SAH) mimics that of SAH. In brain swelling due to acute hypoxic anoxic encephalopathy, the increased attenuation in the basal cisterns and subarachnoid spaces on CT scans is a characteristic finding of acute SAH. PMID- 18410842 TI - Rhabdomyolysis: a lesson on the perils of exercising and drinking. AB - Acute abdomen is a common presentation to the emergency department (ED), accounting for 5% to 10% of ED visits. Of these, 10% require surgery, and 25% go undiagnosed. Usually, most of the cases of undiagnosed abdominal pain are in young women with pelvic etiologies, although occasionally, unusual causes of abdominal pain lead to diagnostic dilemmas and can have adverse clinical outcomes. We present an unusual etiology of abdominal pain in a young man, who presented with acute onset of right lower quadrant pain accompanied by nausea and vomiting. He was an amateur boxer who had recently intensified his training regimen and admitted to binge drinking for several days before presentation. The initial diagnosis was acute appendicitis, but a computed tomographic scan done revealed a normal appendix. Creatine kinase level was then checked and found to be significantly elevated, and a diagnosis of isolated abdominal wall rhabdomyolysis was made. PMID- 18410843 TI - Life-threatening subcutaneous hematoma. AB - Neurofibromatosis is rare genetic disorder that predisposes patients to tumor formation. Although the clinical manifestations are variable, vascular malformations are known to occur. We report a case of life-threatening massive subcutaneous hemorrhage after minor trauma in a patient with neurofibromatosis. Physicians need to be aware that bleeding from vascular malformations, even in the subcutaneous space, may be life threatening. PMID- 18410844 TI - Missed epidural brain abscess after furunculosis. AB - Community-associated methicillin-resistant Staphylococcus aureus (ca-MRSA) has been implicated as a major cause of cutaneous skin infections. Invasive infections from ca-MRSA have also been reported, including endocarditis, pneumonia, and necrotizing fasciitis. We describe a case of a missed ca-MRSA epidural brain abscess in a patient with a recent furunculosis who underwent a lumbar puncture for meningitis workup without a prior head computed tomography. When dealing with invasive infections, physicians need to consider ca-MRSA as a possible cause. PMID- 18410846 TI - Narcolepsy in children: a single-center clinical experience. AB - Although the initial manifestations of narcolepsy in children may differ from those with adult onset, hypersomnia remains the most common presenting sign. This study aimed to (1) describe the clinical and polysomnographic features, and treatment outcomes, of a group of children with narcolepsy, and (2) describe other sleep disorders to be considered in the differential diagnosis of hypersomnia and which may coexist with narcolepsy. A retrospective review of 125 children referred in 1 year for hypersomnia revealed 20 patients (16%) with narcolepsy. Of these, only 15% exhibited cataplexy, 10% experienced hallucinations, and none manifested sleep paralysis. Eighty-five percent of children with narcolepsy had sleep-disordered breathing on polysomnography. Body mass indices of these children were higher than for healthy, age-matched controls subjects. Other polysomnography findings included periodic limb movements of sleep (25%) and parasomnias (5%). The multiple sleep latency test revealed a mean sleep latency of 6.14 minutes, with sleep-onset rapid eye movement periods (median, 2/5 naps). Treatment with modafinil and sodium oxybate provided optimal control of daytime sleepiness. Physicians should routinely screen for hypersomnia in children by obtaining a detailed history and, in appropriate situations, ordering polysomnographic testing to rule out narcolepsy and other causes of hypersomnia. PMID- 18410845 TI - Presentation and diagnosis of mitochondrial disorders in children. AB - The first disorder of mitochondrial function was described by Luft in 1959. Over the ensuing decades, multiple cases of mitochondrial dysfunction were reported, and the term "mitochondrial disorder" arose to describe any defect in the mitochondrial electron transport chain. The sequence of the mitochondrial genome was elucidated in 1981 by Anderson et al., and during the next 20 years, >200 pathogenic point mutations, deletions, insertions, and rearrangements were described. Most of the original cases were adults, and the diagnosis of a mitochondrial disorder in an adult patient became relatively straightforward. Adults present with well-defined "mitochondrial syndromes" and generally carry mitochondrial DNA mutations that are easily identified. Children with mitochondrial disorders are much harder to define. Children are more likely to have a nuclear DNA mutation, whereas the "classic" syndromic findings tend to be absent. This review describes both the varying presentations of mitochondrial disorders and the common laboratory, imaging, and pathologic findings related to children. PMID- 18410847 TI - Magnetic resonance imaging in children with bilateral spastic forms of cerebral palsy. AB - We analyzed the relationship between magnetic resonance image findings in children with bilateral spastic cerebral palsy and its stages of severity in term and preterm children. Magnetic resonance image findings of 102 children (66 male and 36 female) with bilateral spastic cerebral palsy (median age, 2.5 years; range, 3 months to 15 years) were reevaluated. The study group consisted of children with confirmed perinatal asphyxia. Hypoxic-ischemic events were diagnosed in 64% of the children. Significant abnormalities relevant to cerebral palsy were evident on imaging in 85/102 (83%) children (in 77% of term and 93% of preterm children). Enlargement of the ventricles alone (48%) or accompanied by periventricular white-matter damage (25%) was the most frequent finding in term and preterm children, but was more highly expressed in preterm children (P < 0.05). White-matter damage was more often found in preterm children (P < 0.05). Enlargement of the lateral ventricles and periventricular leukomalacia may be attributable to ischemic damage to the neonatal brain. Significant correlations were found between magnetic resonance image findings and severity of cerebral palsy (P < 0.05). Detection of brain abnormalities in children with cerebral palsy may prove useful in prognoses as well as in medical consultations and management. PMID- 18410848 TI - Very early arterial ischemic stroke in premature infants. AB - Early stroke in the premature infant has rarely been described. Presented here are the cases of 23 infants, born between 23 and 35 weeks gestational age, with focal arterial ischemic stroke occurring before 44 weeks gestational age. Ten (43%) were male. Five children (22%) were half of a twin pair; no co-twin died. The most commonly affected territory was the middle cerebral artery territory. Three children with extreme prematurity (< or =26 weeks) had cerebellar infarcts. Twelve children had unilateral or bilateral intraventricular hemorrhages (grade 3 or higher in 8 of the 12). Twelve children had white matter injury: periventricular leukomalacia, hypoxic-ischemic encephalopathy, or both. Most children had multiple comorbidities, and the median neonatal intensive care unit stay was 63 days (range, 14-365). One child died in the neonatal intensive care unit (age 123 days). All 22 survivors were left with disabilities. Seventeen (77%) had cerebral palsy, 10 (45%) had epilepsy, and 17 (77%) had cognitive impairment. Arterial ischemic stroke appears to add to the neurologic disabilities commonly associated with prematurity. PMID- 18410849 TI - Self-concept in adolescents with epilepsy: biological and social correlates. AB - The purposes of this study were to (1) compare self-esteem in teens with epilepsy to the normative mean, and (2) identify which neurologic/epilepsy and social/familial variables are associated with self-esteem. Thirty-seven adolescents (aged 12-18 years) attending a pediatric neurology clinic completed the Piers-Harris 2 Self-Concept Scale, Family Assessment Measure III, Child Attitude to Illness Scale, and a brief questionnaire about current seizure status (frequency, severity, and number of antiepileptic drugs). Neurology clinic charts were reviewed for seizure types, etiology, age at diagnosis, and number of failed therapies. While Total Piers-Harris t score and most subscales did not differ significantly from the normative mean, teens with epilepsy had higher scores on Behavioral Adjustment (P < 0.04) and Physical Appearance and Attributes (P < 0.03). On univariate analysis, number of current antiepileptic drugs (P < 0.05) and Attitude to Illness and Family Function scores (P < 0.02 for both) were significantly associated with self-esteem. On linear regression analysis, only the Family Function score (P < 0.02) and number of antiepileptic drugs (P < 0.05) were associated with total self-concept. We conclude that self-concept in teens with epilepsy is most strongly associated with Family Function. With the exception of current number of antiepileptic drugs used, epilepsy-specific factors are of minimal importance. PMID- 18410850 TI - Discontinuation of medications after successful epilepsy surgery in children. AB - To evaluate the need for antiepileptic drugs after successful epilepsy surgery in pediatric patients, we retrospectively reviewed patients who had epilepsy surgery and were seizure free or had rare nondisabling auras during the first 6 postoperative months. Association between drug discontinuation and seizure recurrence was evaluated using Cox proportional hazards multivariable survival analysis. Medications were withdrawn in 68 of 97 patients, seizure free (or with rare nondisabling auras) for >6 months after surgery; 57 of the 68 (84%) remained seizure free; the other 11 (16%) had seizure recurrence after 68 months (median). Seizure recurrence was controlled with medication in 7 of the 11 patients (3 have rare seizures, 1 frequent auras). Discontinuing medications at <6 mo, compared with later or no withdrawal, had significant risk for seizure recurrence (hazard ratio 5.8; 95% confidence interval 1.8, 17.5; P = 0.003). Of 29 patients who continued drugs, 28 (97%) remained seizure free after 37 months (median). Freedom from seizures 6 months after surgery predicted good outcome (95% seizure free, with or without medication). If discontinuation is offered after 6 months, the majority of patients (84%) can be expected to remain seizure free with no further need for medication. Although seizure breakthrough is possible in a smaller percentage, restarting drugs is likely to restore seizure control. PMID- 18410851 TI - Symptomatic parieto-occipital epilepsy as sequela of perinatal asphyxia. AB - Perinatal hypoxic-ischemic encephalopathy in term infants has been well documented as a cause of selective brain damage over the parasagittal brain regions in cases of partial asphyxia. We describe 10 patients (7 boys, 3 girls) who developed normally after perinatal asphyxia, and who later experienced parieto-occipital epilepsy. We retrospectively reviewed medical records, electroencephalograms, and neuroimaging findings. All patients had a history of perinatal asphyxia requiring intensive care. Subsequent developmental milestones were apparently normal. Seizures started at ages ranging from 10 to 168 months (mean, 72 months). The seizures comprised complex partial seizures with visual symptomatology in 5 cases, complex partial seizures without visual symptomatology in 4 cases, and focal motor seizures with secondary generalization in the remaining case. Brain magnetic resonance imaging demonstrated localized parieto occipital lesions in 9 cases, and normal findings in the remaining one. Electroencephalograms revealed predominantly occipital foci in 8. In all patients, the epileptogenic foci involved either unilateral or bilateral parieto occipital regions. Intelligence quotients and visuospatial cognitive functions were impaired in 7 patients. Seizures were finally controlled in only 4 patients. Symptomatic parieto-occipital epilepsy caused by mild perinatal hypoxic-ischemic encephalopathy is sometimes pharmaco-resistant and accompanied by mild cognitive and visuospatial impairments. PMID- 18410852 TI - Three patients with severe bilateral frontoparietal polymicrogyria. AB - The bilateral frontoparietal polymicrogyria syndrome is characterized anatomically by predominantly frontoparietal diffuse polymicrogyria and clinically by generalized tonic-clonic seizures and mental retardation developing in early childhood. Despite the diffuse nature of the cortical involvement, patients do gain the ability to speak and to walk. In addition, epilepsy can be relatively well controlled with antiepileptic drugs. In three cases of bilateral frontoparietal polymicrogyria, the patients demonstrated consistently similar clinical and neuroimaging characteristics, with more severe structural and developmental abnormalities than those reported previously. The designation proposed here recognizes such cases as a severe form of bilateral frontoparietal polymicrogyria. An ethnic difference or a causal gene other than GPR56 might be responsible for the difference in severity between the classical and the more severe forms of bilateral frontoparietal polymicrogyria. PMID- 18410853 TI - Mycoplasma pneumoniae encephalopathy: recovery after intravenous immunoglobulin. AB - In a case of Mycoplasma pneumoniae encephalopathy in a 7-year-old girl, the microorganism was identified by serologic and pharyngeal-swab polymerase chain reaction methods. Failure to detect or isolate the pathogen in the cerebrospinal fluid and the significant rapid clinical recovery of the patient after intravenous immunoglobulin administration suggest an immune-mediated mechanism. In critically ill patients with M. pneumoniae encephalopathy, prompt intravenous immunoglobulin treatment may be beneficial and can be considered as a primary or alternative therapeutic approach, especially when use of corticosteroids is insecure. PMID- 18410854 TI - Usefulness of multifocal VEP in a child requiring perimetry. AB - We examined the reliability of multifocal visual evoked potentials for evaluating visual-field defects in a child with epilepsy and an arachnoid cyst. Multifocal visual evoked potentials were measured both when perimetry was difficult and several years later, when kinetic perimetry became possible in a child with epilepsy and homonymous hemianopia, as suggested by computed tomography of the brain. The peak latency and amplitude of response waves were used for assessment. The recordings of multifocal visual evoked potentials at both times revealed marked decreases in amplitude in the left visual hemifield. This area of decreased amplitude corresponded to the location of the lesion observed with imaging techniques, and was consistent with the left homonymous hemianopia observed via kinetic perimetry. The objective evaluation of visual-field defects through multifocal visual evoked potentials may be useful in children in whom conventional perimetry is difficult. PMID- 18410855 TI - Dilated perivascular spaces: an informative radiologic finding in Sanfilippo syndrome type A. AB - Mucopolysaccharidosis type IIIA, or Sanfilippo syndrome type A, is a lysosomal storage disorder caused by deficiency of heparan N-sulfamidase, resulting in defective degradation and subsequent storage of heparan sulfate. It is characterized by progressive nervous system involvement. Cribriform changes in the corpus callosum, basal ganglia, and white matter, diffuse high-intensity signal in the white matter, and cerebral atrophy have been described in patients with this disorder. This case report describes a child with Sanfilippo syndrome type A who exhibited fairly mild clinical findings but an unusual magnetic resonance imaging pattern that included multiple moderate-sized cysts (probably enlarged perivascular spaces) within the corpus callosum and an abnormal appearance of the clivus and cervical vertebrae. This case calls attention to the variety of appearances possible with magnetic resonance imaging in Sanfilippo syndrome type A. PMID- 18410856 TI - Autosomal-dominant guanosine triphosphate cyclohydrolase I deficiency with novel mutations. AB - Dopa-responsive dystonia in children, including guanosine triphosphate cyclohydrolase I deficiency, is an important subcategory of treatable dystonia characterized by a dramatic, sustained response to levodopa. Early diagnosis is difficult, however, because of the heterogeneity of the clinical phenotype. We report on two Korean children affected with dopa-responsive dystonia caused by a novel missense mutation of the guanosine triphosphate cyclohydrolase I gene. One child exhibits a novel sporadic mutation, and the other child demonstrates autosomal-dominant inheritance. PMID- 18410857 TI - Postvaricella acute transverse myelitis in a previously vaccinated child. AB - We describe a 14-year-old boy with acute transverse myelitis after breakthrough varicella infection, despite immunization with the Varicella zoster virus vaccine 8 years earlier. He recovered fully after treatment with intravenous corticosteroids and acyclovir. To our knowledge, there are no previously reported cases of postvaricella acute transverse myelitis in vaccinated individuals. Our report emphasizes that the Varicella zoster virus booster vaccine may be necessary to prevent not only acute varicella, but also its postinfectious neurologic complications. PMID- 18410858 TI - Magnetic resonance imaging findings in Alexander disease. PMID- 18410859 TI - The reuse of anesthesia breathing systems: another difference of opinion and practice between the United States and Europe. PMID- 18410860 TI - Emotional intelligence and its correlation to performance as a resident: a preliminary study. AB - STUDY OBJECTIVE: To test the hypothesis that emotional intelligence, as measured by the Bar-On Emotional Quotient Inventory (EQ-I) 125 (Multi Health Systems, Toronto, Ontario, Canada) personal inventory, would correlate with resident performance. DESIGN: Prospective survey. SETTING: University-affiliated, multiinstitutional anesthesiology residency program. PARTICIPANTS: Current clinical anesthesiology years one to three (PGY 2-4) anesthesiology residents enrolled in the University of Pittsburgh Anesthesiology Residency Program. MEASUREMENTS: Participants confidentially completed the Bar-On EQ-I 125 survey. Results of the individual EQ-I 125 and daily evaluations by the faculty of the residency program were compiled and analyzed. MAIN RESULTS: There was no positive correlation between any facet of emotional intelligence and resident performance. There was statistically significant negative correlation (-0.40; P < 0.05) between assertiveness and the "American Board of Anesthesiology essential attributes" component of the resident evaluation. CONCLUSIONS: Emotional intelligence, as measured by the Bar-On EQ-I personal inventory, does not strongly correlate to resident performance as defined at the University of Pittsburgh. PMID- 18410861 TI - Comparing the effect of arginine vasopressin on ear and finger photoplethysmography. AB - STUDY OBJECTIVE: To test whether the relative insensitivity of craniofacial vessels to catecholamines differs in response to arginine vasopressin. DESIGN: Prospective, observational human study. SETTING: University hospital. PATIENTS: 8 ASA physical status I and II women scheduled for elective myomectomy. INTERVENTIONS: Patients underwent elective myomectomy surgery with intrauterine injection of arginine vasopressin. MEASUREMENTS: Finger, ear, and forehead photoplethysmographs were monitored. Changes in the plethysmographic amplitudes were recorded before and after arginine vasopressin injection. MAIN RESULTS: In all subjects, ear photoplethysmographic amplitude (but not oxygen saturation) decreased precipitously (62% +/- 10%; P < 0.001) after arginine vasopressin injection. In contrast, there was no significant decline in the finger signal (4.5% +/- 27%; P = 0.19). The forehead plethysmograph decreased in amplitude, but this finding did not achieve significance (33% +/- 18%; P = 0.18). CONCLUSION: In contrast to prior observations during adrenergic activation, arginine vasopressin induced relatively greater vasoconstriction at the ear and forehead than at the finger. This finding has potential implications with respect to arginine vasopressin's effect on blood flow and indicates that monitoring the ear plethysmographic signal may provide useful information during arginine vasopressin administration. PMID- 18410862 TI - The hemodynamic effects of phenoxybenzamine in neonates, infants, and children. AB - STUDY OBJECTIVES: To evaluate the hemodynamic effects of the long-acting, alpha adrenergic blocker, phenoxybenzamine, in children of different age groups. DESIGN: Retrospective chart review. SETTING: Tertiary-care, congenital cardiac surgery center. MEASUREMENTS: The data of 75 pediatric patients who received phenoxybenzamine while undergoing surgical repair of congenital heart defects on cardiopulmonary bypass (CPB) were studied. Patients were selected in three age groups: younger than one month (n = 25), one to 12 months (n = 25), and one to 5 years (n = 25). All patients received a full dose of phenoxybenzamine 1 mg/kg. Demographics, CPB duration, mean arterial pressure on CPB, mean flow on CPB (normalized to body surface area), and central-to-peripheral temperature gradients were recorded. Systemic vascular resistance index (SVRI) was calculated. MAIN RESULTS: Cardiopulmonary bypass duration was significantly longer in the age group of younger than 1 mo than in the older groups. Mean CPB flow/body surface area was similar in all children (3.45 +/- 0.9, 3.74 +/- 0.69, and 3.48 +/- 0.59 L/min/m2, respectively; P < 0.28). However, mean SVRI was significantly lower in children younger than 1 mo (997.3 +/- 233, 1196.9 +/- 394, and 1168.83 +/- 227 dynes/cm2m5; P < 0.04). Temperature gradient was significantly narrower in patients younger than one month than those who were one to 12 months and one to 5 years at the end of cooling (0.90 degrees C +/- 0.1 degrees C, 1.04 degrees C +/- 3.61 degrees C, 1.4 degrees C +/- 3.07 degrees C; P < 0.001) at end-rewarming and termination of CPB (4.58 degrees C +/- 2.36 degrees C, 6.23 degrees C +/- 4.17 degrees C, 7.32 degrees C +/- 3.46 degrees C; P < 0.02). Multivariate analysis showed that patient age was a significant variable affecting response to phenoxybenzamine, after adjusting for duration of CPB (P = 0.31), mean hematocrit on CPB (P = 0.86), and core cooling temperature (P = 0.34). CONCLUSION: The effect of phenoxybenzamine on SVRI, cooling, and rewarming on CPB varies with age as shown by more profound vasodilatation and narrower temperature gradients. PMID- 18410863 TI - Optimal conditions for Laryngeal Mask Airway insertion in children can be determined by the trapezius squeezing test. AB - STUDY OBJECTIVE: To evaluate the trapezius squeezing test as a criterion of adequate anesthetic depth for Laryngeal Mask Airway (LMA) insertion in children without neuromuscular blocking agents. DESIGN: Prospective, randomized clinical trial. SETTING: Operating room of a university hospital. PATIENTS: 45 ASA physical status I and II children, aged one to 6 years, undergoing minor surgical procedures. INTERVENTIONS: An LMA was inserted in each child after the trapezius squeezing test turned to be negative. MEASUREMENTS: Presence of coughing, gagging, gross purposeful movements, breath holding, laryngospasm, or desaturation during or within one minute of LMA insertion were recorded. Elapsed time of LMA insertion, end-tidal sevoflurane concentration, blood pressure, and heart rate values were also noted. MAIN RESULTS: LMA insertion was successful in 41 patients. Elapsed time to complete insertion was 5.7 +/- 1.9 minutes, and end tidal sevoflurane concentration was 3.6 +/- 1.1 vol%. No marked hemodynamic changes occurred in any child. CONCLUSIONS: The trapezius squeezing test is a reliable clinical indicator to assess adequate anesthetic depth for LMA insertion in children without neuromuscular blocking agents. PMID- 18410864 TI - Efficacy of lornoxicam for acute postoperative pain relief after septoplasty: a comparison with diclofenac, ketoprofen, and dipyrone. AB - STUDY OBJECTIVES: To compare the efficacy of injectable lornoxicam with diclofenac, ketoprofen, and dipyrone for acute postoperative pain. DESIGN: Prospective, randomized, placebo-controlled, double-blind study. SETTING: University hospital. PATIENTS: 200 ASA physical status I patients who were scheduled for elective septoplasty with general anesthesia. INTERVENTIONS: Patients were divided into 5 groups according to the intramuscularly administered analgesic drug: lornoxicam 8 mg (twice daily), diclofenac 75 mg (twice daily), ketoprofen 100 mg (twice daily), dipyrone 1 g (three times daily), and placebo (twice daily). MEASUREMENTS: Pain intensity was evaluated with a 0 to 100 mm Visual Analog Scale (VAS) at the 2nd, 4th, 6th, 8th, 12th, 16th, 20th, and 24th hour postoperatively. Intramuscular pethidine 1 mg/kg was administered to patients requiring additional analgesia, and treatment-related adverse effects were noted. MAIN RESULTS: Pethidine requirement was found to be significantly higher in the placebo group (1.8 mg/kg per 24 hours; 95% confidence interval, 1.5 2.2) than in the other groups (P = 0.001). No significant difference in opioid requirement was found among the treated groups (P > 0.05). Postoperative VAS scores were significantly lower at specific hours in the treatment groups when compared with placebo group (P < 0.05). No statistically significant difference in adverse effects was found among the groups studied (P > 0.05). CONCLUSIONS: Efficacy of lornoxicam in the management of acute postoperative pain was not superior to that of other nonopioid analgesics used in this study. PMID- 18410865 TI - Reusable Laryngeal Mask Airways can be used more than 40 times. AB - STUDY OBJECTIVE: To compare the mechanical properties of classic, reusable Laryngeal Mask Airways (LMAs) that have been used more than 100 times with one unused LMA. DESIGN: Laboratory testing of devices used clinically. SETTING: Metropolitan university hospital. MEASUREMENTS: Cuff and tube specimens from LMAs that had been used at least 100 times were tested for elongation, tensile strength, stiffness, and tear strength using standard American Society of Testing and Materials protocols. Samples from an unused LMA were analyzed in the same manner for comparison. MAIN RESULTS: Tensile strength of the cuff samples was found to increase by approximately 25%. There was an increase in cuff stiffness and decreased tear strength similar to the manufacturer-reported trends. Cuff elongation decreased by 30%. Results for the LMA tube differed depending on whether the samples were taken in the machine direction or transverse direction. Tensile strength decreased by 30% in machine direction and, on average, very little in transverse direction. CONCLUSIONS: The material in reusable classic LMAs does not lose its strength after 100 uses to the extent that its manufacturer claims. At least 100 uses may be considered safe for these devices. PMID- 18410866 TI - Brain tumors may alter the relationship between bispectral index values and propofol concentrations during induction of anesthesia. AB - STUDY OBJECTIVE: To compare propofol-predicted effect-site concentrations (PropCe) and bispectral index (BIS) of the electroencephalogram during induction of anesthesia in patients with small brain tumors and to analyze BIS and PropCe at loss of consciousness (LOC). DESIGN: Prospective investigation. SETTING: Operating theater of a university hospital. PATIENTS: 26 ASA physical status I and II patients, 13 of whom were scheduled for nontumor spinal surgeries, and the other 13, for brain surgery for small brain tumor removal. INTERVENTIONS: Anesthesia was induced with a propofol 1% constant infusion rate of 200 mL/h until LOC. MEASUREMENTS: BIS, PropCe, heart rate, and mean arterial pressure were analyzed at the beginning of the propofol infusion and every 30 seconds during induction. MAIN RESULTS: The BIS values were significantly higher in patients with brain tumors in the period from 150 to 210 seconds, with PropCe similar to patients without brain tumors. Loss of consciousness occurred 3.6 +/- 0.8 minutes in patients without brain tumors and 3.9 +/- 0.7 minutes in patients with brain tumors. No differences were observed between groups in the time to LOC (3.6 +/- 0.8 in group 1 vs 3.9 +/- 0.7 in group 2) or in BIS at LOC (48.7 +/- 11.4 in group 1 vs 58.6 +/- 21.7 in group 2). CONCLUSIONS: For similar propofol concentrations, patients with small brain tumors show higher BIS values on induction of anesthesia with propofol. PMID- 18410867 TI - Preoperative beta-blocker use: impact of national guidelines on clinical practice. AB - STUDY OBJECTIVE: To determine if recommendations regarding perioperative beta blocker therapy were followed by an increase in the number of eligible presurgical patients receiving beta-blockers and the number achieving the recommended heart rate (HR <60 beats per minute [bpm]). DESIGN: Retrospective, observational study. SETTING: Tertiary-care teaching hospital. MEASUREMENTS: The records of all 718 patients who underwent elective vascular surgery or coronary artery bypass grafting between January 2001 and March 2002 (pre-guideline) and those who did so between April 2002 and September 2003 (post-guideline) were reviewed. Percentage of eligible patients who received beta-blockers preoperatively and the target HR achieved in pre-guideline versus post-guideline patients were recorded. Differences were assessed using the unpaired t test and chi2 analysis. A P value of less than 0.05 is reported. MAIN RESULTS: Fifty percent of the post-guideline patients in the vascular surgery group were receiving beta-blockers at the time of preanesthetic evaluation versus 48% of pre guideline patients (P = nonsignificant [NS]). Mean HR in the vascular surgery post-guideline beta-blocker group (70 +/- 14 bpm) was higher than in the pre guideline beta-blocker group (65 +/- 11 bpm) (P < 0.01). Only 22% of those vascular surgery patients in the post-guideline group who were taking beta blockers achieved the target HR of less than 60 bpm versus 29% of the vascular surgery patients taking beta-blockers in the pre-guideline group (P = NS). In the coronary artery bypass grafting group, 80% of post-guideline patients received beta-blocker before anesthesia assessment versus 75% of pre-guideline patients (P = NS). Mean HR in the post-guideline beta-blocker group (67 +/- 15 bpm) was similar to the pre-guideline beta-blocker group (64 +/- 13 bpm) (P = NS). Only 28% of the post-guideline patients who were receiving beta-blockers achieved the target HR of less than 60 bpm, which was not significantly different from the 17% achieved in the pre-guideline group (P = NS). CONCLUSION: At our institution, preoperative beta-blocker use was not significantly changed by publication of the recommendations. PMID- 18410868 TI - Neurogenic pulmonary edema in a fatal case of subarachnoid hemorrhage. AB - Neurogenic pulmonary edema (NPE) is caused by a variety of central nervous system lesions and may appear as a subclinical complication. The fulminant form of NPE is always life-threatening. Many pathophysiologic mechanisms have been implicated in the development of NPE, but the exact interaction remains unknown. We report a case of a fulminant NPE with fatal consequences associated with a subarachnoid hemorrhage. Treatment focuses on ventilatory support and measures to reduce intracranial pressure. PMID- 18410869 TI - Ventilatory management of the patient with hyperimmunoglobulinemia E (Job) syndrome. AB - Hyperimmunoglobulinemia E (Job syndrome) is a primary immunodeficiency that features abscesses of the skin, lung, and viscera; pneumonia; and elevated levels of serum immunoglobulin E. Pulmonary infections lead to lung abscesses, pneumatoceles, and bronchiectasis. We report the ventilatory management of a patient with Job syndrome. PMID- 18410870 TI - A novel view for visualizing a left pulmonary artery thromboembolus with intraoperative transesophageal echocardiography. AB - Visualization of thromboembolic material in the pulmonary artery is often difficult on transesophageal echocardiography, especially in the left pulmonary artery, because of the position of the left main bronchus. We present a case in which thromboembolic material within the midleft pulmonary artery was incidentally diagnosed using additional, modified transesophageal echocardiography views, in a patient undergoing mitral valve repair. PMID- 18410871 TI - A case of refractory intraoperative hypotension treated with vasopressin infusion. AB - A 56-year-old man, treated with an angiotensin II receptor antagonist for hypertension, presented for placement of a cochlear implant during general anesthesia. Intraoperatively, there was profound hypotension that was resistant to decreasing the anesthetic depth, fluid administration, as well as bolus doses of phenylephrine, ephedrine, and epinephrine. Hypotension was eventually successfully treated with a vasopressin infusion (0.06 U/min). Vasopressin may be a useful agent in such scenarios because its effect is not dependent on either adrenergic or angiotensin receptors, both of which may be affected by angiotensin II receptor antagonists. PMID- 18410872 TI - A possible perianesthetic serotonin syndrome related to intrathecal fentanyl. AB - Serotonin syndrome occurs with selective serotonin reuptake inhibitors, opioids, and other serotonergic agents. We describe a possible serotonin syndrome related to intrathecal fentanyl in a patient taking multiple drugs and substances such as ergot alkaloids, marijuana, methylenedioxy-N-methylamphetamine, and ephedrine. PMID- 18410874 TI - Trust but verify: cooperation cannot mean abdication in the operating room. PMID- 18410873 TI - Occurrence of intermittent Wolff-Parkinson-White syndrome during intravenous sedation. AB - Patients with intermittent Wolff-Parkinson-White (WPW) syndrome, defined as intermittent loss of the delta waves, can show occasional conduction through the accessory pathway. WPW syndrome often causes paroxysmal supraventricular tachycardia or atrial fibrillation. However, it may be difficult to identify the abnormalities preoperatively because of their only intermittent occurrence. We report a case in which exogenously administered epinephrine and an autonomic imbalance may have precipitated the abrupt occurrence and disappearance of the delta waves. PMID- 18410875 TI - Recurrent fever, chills, and arthralgia with local anesthetics containing epinephrine-metabisulfite. PMID- 18410876 TI - Fluid restriction in the management of postobstructive pulmonary edema: wise or otherwise? PMID- 18410877 TI - The Laryngeal Mask Airway may help to protect the lungs and control bleeding in a patient with significant pharyngeal bleeding. PMID- 18410878 TI - Ultrasound-guided transabdominus plane block. PMID- 18410879 TI - The use of a Laryngeal Mask Airway in cardiac anesthesia and intra-arterial injection of rocuronium. PMID- 18410880 TI - How is body mass index protective in maintenance dialysis? PMID- 18410881 TI - Public health approach to addressing hyperphosphatemia among dialysis patients. AB - Elevated serum phosphorus levels are a major source of morbidity and mortality for the 350,000 Americans receiving chronic dialysis treatment. Despite the widespread application of medical and behavioral interventions, the prevalence of hyperphosphatemia remains exceedingly high. At first glance, a public health perspective may seem inappropriate for addressing a disorder of mineral metabolism among patients receiving a life-sustaining treatment. However, we analyzed this topic from a public health perspective and identified many opportunities to improve the management of hyperphosphatemia, including (1) media and cultural messages about food, (2) the availability of appropriate foods and medications, (3) physical structures such as the location of products in grocery stores, and (4) social structures such as food-labeling laws. PMID- 18410882 TI - Impact of prealbumin levels on mortality in patients with acute kidney injury: an observational cohort study. AB - OBJECTIVE: We evaluated the prognostic value of prealbumin (transthyretin) levels in patients with acute kidney injury (AKI). STUDY DESIGN: This was a longitudinal, single-center, observational cohort study. SETTING: The setting was a university-affiliated hospital. PARTICIPANTS: One hundred and sixty-one consecutive patients with AKI matched at least one of the Risk, Injury, Failure, Loss, End Stage (RIFLE) criteria for increased serum creatinine, and had requested a nephrology consultation between April 2000 and March 2006. METHODS: We conducted a computer-assisted review of all cases of AKI seen by a nephrologist between April 2000 and March 2006. We used Cox proportional hazards models to estimate the risk of in-hospital mortality associated with serum prealbumin level at the beginning of the nephrology consultation. RESULTS: Clinicopathologic variables were compared between patients with a serum prealbumin level <11 mg/dL (the exposed group; cutoff point, median) and patients with a serum prealbumin level > or =11 mg/dL (the control group). In-hospital mortality rates associated with prealbumin levels were 45.0% for <11 mg/dL, and 19.8% for > or =11 mg/dL (P = .001). After adjusted analysis, the presence of a serum prealbumin level <11 mg/dL was significantly associated with increased in hospital mortality (hazard ratio, 2.10; 95% confidence interval, 1.08 to 4.08), compared with patients with the same Liano scoring, the same RIFLE classes, and the same treatment for AKI, but with a serum prealbumin level > or =11 mg/dL. In addition, each increment of 5 mg/dL of serum prealbumin was associated with an adjusted 29% decrease of in-hospital mortality (hazard ratio, 0.71; 95% confidence interval, 0.52 to 0.96). CONCLUSIONS: Serum prealbumin levels <11 mg/dL were strongly associated with a higher risk of death, independent of AKI severity, comorbid illnesses, serum C-reactive protein levels, and other possible confounders. The measurement of serum prealbumin levels may be an inexpensive and useful tool in the evaluation of the risk profiles of AKI patients. PMID- 18410883 TI - Positive effects of protein restriction in patients with chronic kidney disease. AB - OBJECTIVES: The potential benefit or harm of low-protein diets (LPDs) for patients with chronic kidney disease has been debated. This study sought to investigate the effects of treatment with LPDs on nutritional markers, morbidity, and survival during subsequent dialysis. A second objective was to evaluate the effect of LPDs on renal function and the start of dialysis. DESIGN: This was a retrospective study of medical records. SETTING: The setting was an outpatient nephrology and dialysis clinic. PATIENTS: One-hundred twenty-two renal patients were recruited from the central dialysis registry of one clinic. The patients had been followed by a nephrologist for > or =6 months before dialysis. Sixty-one patients were treated with LPDs, and an equal number of control patients not treated with LPDs were matched for sex, age, dialysis modality, diabetes, and start of dialysis. MAIN OUTCOME MEASURES: Main outcome measures included weight and weight change, serum albumin, glomerular filtration rate, morbidity, and mortality. RESULTS: There was less mean weight loss in the LPD group the year before dialysis (0.14 kg/month, compared with the control group at 0.36 kg/month, P < .05). The level of serum albumin was higher in the LPD group at the start of dialysis (P < .01). The mean rate of progression during the 6 months before dialysis was lower in the LPD group (4.1 mL/min/year) than in the control group (13.4 mL/min/year) (P < .001). The LPD group had fewer days of hospitalization at the start of dialysis than the control group (8.2 vs 15.4 days, respectively, P < .01). There was no difference in mortality between groups 1, 2, or 5 years after starting dialysis. CONCLUSIONS: Low-protein diets can reduce patient morbidity, preserve renal function, relieve uremic symptoms and improve nutritional status. The results suggest that LPDs can postpone the start of dialysis for 6 months, and entail substantial cost-savings. Low-protein diets should be used more generally in the renal community. PMID- 18410884 TI - Assessment of nutritional status among ESRD patients in Jordanian hospitals. AB - OBJECTIVE: Our objective was to assess nutritional status and compare quality of treatment among hemodialysis patients in public and private hospitals in Jordan. DESIGN: We utilized a cross-sectional survey. SETTING: Our setting involved hospital hemodialysis units. PATIENTS: This study was undertaken in five large Jordanian hospitals between 2004 and 2005. One hundred and eighty participants diagnosed with end-stage renal failure (ESRD) were enrolled. These participants (91 women and 89 men) who underwent hemodialysis treatment were recruited using a convenience sampling technique. Data from participants who received hemodialysis treatment in public hospital settings were compared with equivalent data from participants treated in private hospital settings. MAIN OUTCOME MEASURES: Subjective global assessment (SGA), anthropometry, and biochemical measurements were used as evaluative tools. RESULTS: In the anthropometric measurement of triceps skinfold thickness (TSF), we found a statistically significant difference (P < .05) between participants treated in the two hospital settings. In addition, approximately 62% of all participants, regardless of treatment site, were found to be moderately to severely malnourished. Anthropometric measurements showed some significant increase between prehemodialysis and posthemodialysis weight and body mass index in private hospitals; this was not unexpected. There were no statistically significant differences in the measured mean levels of eight different biochemical parameters, with the exception of plasma phosphorus and sodium levels. CONCLUSIONS: The prevalence of malnutrition and the quality of treatment in our two groups of participants were similar. PMID- 18410885 TI - Relevance of nutritional route and intercellular adhesion molecule-1 in patients with acute renal failure and its prognostic implications. AB - OBJECTIVE: Nutritional support and the route of nutrition are important conditions for patients with acute renal failure (ARF) in intensive care units (ICUs). Enteral nutrition (EN) is the primary route of nutrition in these patients because of a lower rate of complications. A lack of enteral feeding was reported to increase intercellular adhesion molecule-1 (ICAM-1) in experimental models. Serum soluble ICAM-1 (sICAM-1) level is an independent predictor of mortality in predialysis patients. However, the effect of nutritional route on serum ICAM-1 level is unknown in ARF patients. The aim of this study was to investigate the relationship between route of nutrition and serum ICAM-1 level and its prognostic implications in ICU ARF patients. METHODS: In total, 64 ICU patients with ARF were assessed according to their clinical features, route of nutrition, laboratory parameters, serum sICAM-1 levels, presence of infection, Acute Physiology and Chronic Health Evaluation (APACHE) III scores, and outcomes on their first nephrology consultation day. RESULTS: Thirty-two patients died during the follow-up period. The mortality rate and infection rate were higher in the parenteral nutrition (PN) group compared with the EN group (64% vs 42%, P = .05, and 84% vs 64%, P = .05, respectively). The route of nutrition influenced the serum sICAM-1 level. Parenteral nutrition was associated with a higher serum sICAM-1 level compared to EN (434 ng/mL [range 255 to 1,240] vs 217 ng/mL [range 123 to 296], respectively, P = .0004). The APACHE III score was found to be an independent prognostic factor among the parameters of nutritional route, presence of infection, serum albumin level, and serum sICAM-1 level. CONCLUSIONS: Patients with ARF as supported by PN had a lower serum albumin level, and a higher APACHE III score, sICAM-1 level, and mortality and infection rate. Serum sICAM-1 levels did not independently predict mortality in the present set of ARF patients. PMID- 18410886 TI - Impact of vitamin D status and obesity on C-reactive protein in kidney-transplant patients. AB - OBJECTIVE: We examined whether vitamin D status and obesity are associated with low-grade systemic inflammation, as assessed by serum concentrations of C reactive protein (CRP) in an adult population of kidney-transplant patients. DESIGN: This was a single-center, cross-sectional study. SETTING AND PATIENTS: Data were collected between December 2005 and April 2006 from 161 adult (aged >18 years) kidney-transplant patients (mean age, 53.1 years; SD, 11.5 years; females/males, 78/83), with a median kidney-graft age of 7.0 years and serum CRP levels < or =10 mg/L. METHODS: Vitamin D status was assessed by serum concentrations of 25-hydroxyvitamin D (25-OHD) and 1,25-dihydroxyvitamin D (1,25(OH)(2)D). The presence of low-grade systemic inflammation was assessed by serum CRP. Selected lifestyle factors and anthropometric variables were determined in a subgroup of patients (n = 90). Bivariate correlation and multiple regression analyses were performed. RESULTS: Hypovitaminosis D (ie, S-25-OHD < or =75 nmol/L) was present in 73% of females, and 89% of males. The median level of serum CRP was 1.7 mg/L. Vitamin D status was not significantly associated with serum CRP. In the patient subgroup, indices of obesity (body mass index, body weight, and fat mass) correlated positively, and lean body mass correlated negatively, with serum CRP. Only fat mass remained significantly associated with serum CRP in multiple regression analysis. CONCLUSIONS: No impact of vitamin D status on low-grade systemic inflammation was found. Fat mass correlated positively with CRP, suggesting that obesity may increase the risk of cardiovascular disease and chronic allograft rejection in kidney-transplant patients. PMID- 18410887 TI - Acute rise of omega-3 polyunsaturated fatty acids during hemodialysis treatment. AB - OBJECTIVE: Hemodialysis patients have an extremely high rate of cardiac arrhythmia-induced sudden cardiac death, although the risk during the hemodialysis procedure is relatively low. A higher blood content of long-chain omega-3 polyunsaturated fatty acids (PUFAs) is believed to reduce the risk of sudden cardiac death. We performed this study to measure the effect of a single standard hemodialysis treatment on plasma and erythrocyte omega-3 PUFA levels in chronic hemodialysis patients. DESIGN: This was a prospective, observational study. SETTING: The study was performed in one outpatient hemodialysis unit. PATIENTS: Study subjects were all chronic, stable hemodialysis patients. INTERVENTIONS: There were no interventions. MAIN OUTCOME MEASURES: Plasma and erythrocyte fatty-acid levels were measured before and immediately after a hemodialysis session. RESULTS: Plasma levels of long-chain PUFAs, including the omega-3 fatty acids of interest, all rose, whereas those of shorter-chain or more saturated fatty acids either remained unchanged or fell. A similar trend was seen in erythrocytes, though the results did not reach statistical significance. CONCLUSIONS: The hemodialysis procedure induces acute increases of long-chain omega-3 PUFAs in the blood. This effect may help explain why malignant cardiac arrhythmias occur relatively infrequently during hemodialysis. PMID- 18410888 TI - Bioelectric impedance predicts total body water, blood pressure, and heart rate during hemodialysis in children and adolescents. AB - OBJECTIVE: The purposes of our study were to: (1) assess if changes in the volemic status of children and adolescents over the course of standard dialysis could be observed using bioelectric impedance (BIA); and (2) evaluate whether the variability of blood pressure (systolic blood pressure, SBP; diastolic blood pressure, DBP) and heart rate (HR) could be explained by independent variables from BIA data. DESIGN: We used a randomized, single-blinded treatment and repeated-measures design. SETTING: This study took place at the DaVita Children's Dialysis Center (Chicago, IL). PATIENTS: There were 7 subjects, aged 10 to 16 years. INTERVENTION: Two identical standard hemodialysis (HD) sessions were completed, with data collected five times during each HD session: pre-HD, intra HD (hours 1, 2, and 3), and post-HD. Endpoints included total body water (TBW), resistance (R), reactance (Xc), bioimpedance vector |Z|, supine and sitting SBP, DBP, and HR. Standing SBP, DBP, and HR were collected pre-HD and post-HD. RESULTS: No differences were observed in TBW between HD sessions for all subjects. However, TBW decreased throughout the HD sessions for all subjects (although no significant differences were seen between hour 3 and post-HD). Reactance (representative of extracellular water) correlated with supine, sitting, and standing SBP (r = 0.55, 0.59, and 0.51, respectively; P < or = .008). The bioimpedance vector increased beginning at hour 1 (P < .001), reflective of a decline in tissue hydration over the course of HD. CONCLUSIONS: Weight gain in end-stage kidney disease patients is largely fluid. Thus, the use of BIA during HD may aid in the prediction of cardiovascular instability before the development of symptoms, because intravascular hypovolemia and hypotension can result from excessive ultrafiltration below the critical dry weight. In addition, BIA explains, in part, the variability of SBP, DBP, and HR during HD. We suggest that our data also demonstrates the delay in mobilization of fluid from the interstitial space for plasma refill, as evidenced by the delayed change in |Z| over HD. Bioelectric impedance is useful for explaining changes in volemic status and, in part, the variability of SBP, DBP, and HR during HD. PMID- 18410889 TI - Resting energy expenditure of children and adolescents undergoing hemodialysis. AB - OBJECTIVE: We compared the resting energy expenditure (REE) of clinically stable children and adolescents on hemodialysis with that of healthy individuals, to investigate whether the current energy recommendations from the National Kidney Foundation/Kidney Disease Outcomes Initiative (NKF/DOQI) are adequate. PATIENTS AND METHODS: A cross-sectional study was conducted in 25 children and adolescents (15 male, SD 12.3 +/- 3.1 years old) undergoing hemodialysis (study group) under stable clinical conditions. Patients were pair-matched by sex and age with 25 healthy individuals (control group). The REE was measured by indirect calorimetry. Nutritional status was evaluated by body mass index Z-score, and growth deficit was evaluated by height-for-age Z-score. Body composition was assessed by dual-energy X-ray absorptiometry. The total energy expenditure (TEE) was estimated by multiplying the REE by the physical activity factor. The recommended dietary allowance (RDA) for daily energy intake was compared with the estimated TEE in the study group, considering children's age-specific height. RESULTS: All anthropometric parameters were significantly lower in the study group. Malnutrition and stunting were found in up to 56% and 60%, respectively, of the children in the study group. The absolute REE was significantly lower in the study group than in the control group (1,067 +/- 191 kcal/day vs 1,372 +/- 290 kcal/day, respectively, P < .01). However, when the REE was adjusted for its main determinant, ie, lean body mass, no significant difference was observed between groups (P = .12). In the study group, the RDA for energy was above the TEE in all age groups, and the mean percentage of RDA/TEE was 124.2% +/- 14.2%. CONCLUSIONS: Children and adolescents on hemodialysis had an important deficit of energy and protein stores. The similar adjusted REE found between hemodialysis patients and their matched healthy pairs suggests that under stable clinical conditions, the REE of these subjects is normal. Moreover, the energy recommendation set by he NKF/DOQI is appropriate for this population. PMID- 18410890 TI - Let's end the debate on "which weight?". PMID- 18410891 TI - The clinical outcome of Hurler syndrome after stem cell transplantation. AB - Hurler syndrome (HS) is a severe inborn error of metabolism causing progressive multi-system morbidity and death in early childhood. At present, stem cell transplantation (SCT) is the only available treatment that can prevent central nervous system disease progression in HS patients. Although SCT has been shown to be effective for several important clinical outcome parameters, the reported clinical outcome after successful SCT is variable among HS patients and there are still some major limitations. This review will focus on the clinical outcome of HS patients after successful SCT, with particular emphasis on the long-term outcome and complications. In addition, factors that are suggested to contribute to the variable outcome are outlined, as well as the limitations of SCT in HS patients. PMID- 18410893 TI - Preengraftment serum C-reactive protein (CRP) value may predict acute graft versus-host disease and nonrelapse mortality after allogeneic hematopoietic stem cell transplantation. AB - In a mouse model, inflammatory cytokines play a primary role in the development of acute graft-versus-host disease (aGVHD). Here, we retrospectively evaluated whether the preengraftment C-reactive protein (CRP) value, which is used as a surrogate marker of inflammation, could predict posttransplant complications including GVHD. Two hundred twenty-four adult patients (median age, 47 years; range: 18-68 years) underwent conventional stem cell transplantation (CST, n = 105) or reduced-intensity stem cell transplantation (RIST, n = 119). Patients were categorized according to the maximum CRP value during neutropenia: the "low CRP" group (CRP < 15 mg/dL, n = 157) and the "high-CRP" group (CRP >or= 15 mg/dL, n = 67). The incidence of documented infections during neutropenia was higher in the high-CRP group (34% versus 17%, P = .004). When patients with proven infections were excluded, the CRP value was significantly lower after RIST than after CST (P = .017) or after related than after unrelated transplantation (P < .001). A multivariate analysis showed that male sex, unrelated donor, and HLA mismatched donor were associated with high CRP values. The high-CRP group developed significantly more grade II-IV aGVHD (P = .01) and nonrelapse mortality (NRM) (P < .001), but less relapse (P = .02). The present findings suggest that the CRP value may reflect the net degree of tissue damage because of the conditioning regimen, infection, and allogeneic immune reactions, all of which lead to subsequent aGVHD and NRM. PMID- 18410892 TI - The allogeneic effect revisited: exogenous help for endogenous, tumor-specific T cells. AB - The "allogeneic effect" refers to the induction of host B cell antibody synthesis or host T cell cytotoxicity, including tumoricidal activity, by an infusion of allogeneic lymphocytes. We show that treatment of mice with cyclophosphamide (Cy) followed by CD8(+) T cell-depleted allogeneic donor lymphocyte infusion (Cy + CD8(-) DLI) induces regression of established tumors with minimal toxicity in models of both hematologic and solid cancers, even though the donor cells are eventually rejected by the host immune system. The optimal antitumor effect of Cy + CD8(-) DLI required the presence of donor CD4(+) T cells, host CD8(+) T cells, and alloantigen expression by normal host but not tumor tissue. The results support a model in which a donor CD4(+) T cell-mediated graft-versus-host (GVH) reaction effectively awakens antitumor immunity among Cy-resistant host CD8(+) T cells. These events provide the cellular mechanism of the "allogeneic effect" in antitumor immunity. Cy + CD8(-) DLI may be an effective and minimally toxic strategy for awakening the host immune response to advanced cancers. PMID- 18410894 TI - CD134-allodepletion allows selective elimination of alloreactive human T cells without loss of virus-specific and leukemia-specific effectors. AB - Graft-versus-host disease (GVHD) remains a frequent and severe complication of allogeneic stem cell transplantation (SCT). One approach to reducing alloreactivity is to deplete the graft of alloreactive T cells. Global T cell depletion results in poor immune reconstitution with high mortality from viral infections and disease relapse. Therefore, an approach to selectively deplete alloreactive T cells without compromising other responses would be highly beneficial. We undertook studies to identify an inducible activation marker expressed on alloreactive effector T cells following culture with HLA-mismatched allostimulators. Compared to other markers, CD134 was superior because of its negative baseline expression and rapid upregulation after activation. Depletion of CD134(+) cells from responder populations dramatically reduced specific alloreactivity as determined by reduction of helper T cell precursor frequencies below the threshold predicting development of clinical GVHD while retaining responses to third-party alloantigens. CD134-allodepleted populations retained effectors specific for the Wilms' tumor (WT1) leukemia antigen as determined by WT1 specific pentamers, and CMV-specific effectors as determined by CMV-specific pentamers and CMV-specific ELISpot. Thus, use of CD134-allodepleted grafts may improve allogeneic SCT by reducing GVHD without loss of pathogen-specific and leukemia-specific immunity. PMID- 18410895 TI - Sirolimus in combination with cyclosporine or tacrolimus plus methotrexate for prevention of graft-versus-host disease following hematopoietic cell transplantation from unrelated donors. AB - In 2 consecutive prospective clinical trials, we evaluated the efficacy of sirolimus together with a calcineurin inhibitor (cyclosporine or tacrolimus) and low-dose methotrexate for prevention of graft-versus-host disease (GVHD) after unrelated hematopoietic cell transplantation (HCT). Nine patients received sirolimus with cyclosporine, and 17 received sirolimus with tacrolimus. The incidence of grade II-IV GVHD was 77%, with the median onset at day 7 after HCT. Because of toxicity, administration of sirolimus was discontinued earlier than planned in 11 patients, but after the onset of GVHD. Three patients developed renal failure requiring hemodialysis. Accrual in both studies was terminated because of lack of efficacy. In these studies, the addition of sirolimus to regimens containing a calcineurin inhibitor and methotrexate appeared to cause toxicity and provided no detectable improvement in preventing GVHD. PMID- 18410896 TI - Similar and promising outcomes in lymphoma patients treated with myeloablative or nonmyeloablative conditioning and allogeneic hematopoietic cell transplantation. AB - We compared the outcomes of 141 consecutive patients who received allogeneic transplantation with either myeloablative (MA) or nonmyeloablative/reduced intensity (NMA) conditioning for non-Hodgkin and Hodgkin lymphoma at the University of Minnesota. All patients were transplanted between 1997 and 2004. NMA transplant recipients were older and received umbilical cord blood grafts more frequently (MA: 6 [9%]; NMA: 33 [43%], P < .001). NMA patients had more advanced disease and 30 (39%) patients had undergone prior autologous transplantation. The 4-year overall survival (OS) (MA: 46% versus NMA: 49%; p = .34) and the 3-year progression-free survival (PFS) (MA: 44% versus NMA: 31%; P = 0.82) were similar after MA or NMA conditioning. However, MA conditioning resulted in significantly higher 1-year treatment-related mortality (TRM) (MA: 43% versus NMA: 17%; P < .01) but a lower risk of relapse at 3 years (MA: 11% versus NMA: 36%; P < .01). We conclude that similar transplant outcomes are achieved after allogeneic hematopoietic stem cell transplantation using MA conditioning in younger patients and NMA conditioning in older patients or those with prior autologous transplantation not eligible for MA conditioning. Modifications to refine patient assignment to the preferred conditioning intensity and reduce relapse risks with NMA approaches are needed. PMID- 18410897 TI - Neuropotency of human mesenchymal stem cell cultures: clonal studies reveal the contribution of cell plasticity and cell contamination. AB - Various studies have shown neuropotency of bone marrow-derived human mesenchymal stem cells (hMSC) based on the appearance of cells with neural phenotype before or after neural induction protocols. However, to date, it is unclear which mechanisms account for this observation. We hypothesized that neural phenotypes observed in hMSC cultures can be because of both intrinsic cell plasticity and contamination by cells of neural origin. Therefore, we characterized 38 clones from hMSC cultures by assessing their adipogenic/osteogenic potential with specific mesenchymal differentiation protocols, and their molecular neural phenotype by RT-PCR analysis before and after exposure to a defined neural stem cell (NSC) medium for 8 days (neural protocol). We found 33 clones with mesenchymal potential and 15 of them also showed a neural phenotype. As neural phenotypes were maintained during the neural protocol, this suggested neural cell plasticity in 39% of all clones through pluripotency. Importantly, we were able to induce neural phenotypes in 11 of mesenchymal clones applying the neural protocol, demonstrating neural cell plasticity in 29% of all clones through the mechanism of transdifferentiation. Finally, 2 of 5 nonmesenchymal clones (5% of all clones) displayed a neural phenotype indicating neural cell contamination of hMSC cultures. In conclusion, we found 2 different ways of neuropotency of hMSC cultures: cell plasticity and cell contamination. PMID- 18410898 TI - Hematopoietic stem cell transplantation in adults with acute myeloid leukemia. AB - Hematopoietic stem cell transplantation (HSCT) is an integral part of the treatment of many patients with acute myeloid leukemia (AML). Despite extensive study, the appropriate role and timing of allogeneic and autologous transplantation in AML are poorly defined. This review critically analyzes the extensive literature, focusing on the recent advances, and provides practical recommendations for the use of HSCT in AML. PMID- 18410899 TI - Excellent outcome of allogeneic hematopoietic stem cell transplantation using a conditioning regimen with medium-dose VP-16, cyclophosphamide and total-body irradiation for adult patients with acute lymphoblastic leukemia. AB - We retrospectively evaluated the outcomes of 37 adult patients with acute lymphoblastic leukemia (ALL) undergoing allogeneic hematopoietic stem cell transplantation (allo-SCT) conditioned with medium-dose VP-16 (VP, 30 mg/kg), cyclophosphamide (CY, 120 mg/kg), and fractionated total-body irradiation (TBI, 12 Gy) (medium-dose VP/CY/TBI). The median age of the patients was 26 years. Thirteen patients underwent transplantation from HLA-matched related donors (MRD), 18 patients underwent transplantation from HLA-matched unrelated donors (MUD), and 6 patients underwent transplantation from HLA-mismatched donors (MMD). Thirty-two patients received bone marrow and 4 patients received peripheral blood stem cells. Ten patients were Philadelphia chromosome-positive (Ph(+)) and 35 patients were in complete remission (CR) at transplantation. All of the patients achieved engraftment, and grade 3 organ toxicity before engraftment occurred in 27 patients. Grade II-III acute graft-versus-host disease (GVHD) and chronic GVHD (cGVHD) occurred in 15 and 18 patients, respectively. No patient developed grade IV acute GVHD (aGVHD) or died of GVHD. At median follow-up of 35.1 months, 32 patients were alive and all Ph(+) patients were alive. Three patients died of relapse and 2 died of transplant-related mortality (TRM). The actuarial 3-year overall survival (OS) rate, relapse rate, and TRM rate were 89.2%, 8.1%, and 5.4%, respectively. Non-CR at transplantation, MRD, and no aGVHD were significant adverse prognostic factors for survival. Medium-dose VP/CY/TBI for adult ALL patients was associated with lower relapse rate and no increase in toxicity, resulting in better survival. PMID- 18410900 TI - IV busulfan dose individualization in children undergoing hematopoietic stem cell transplant: limited sampling strategies. AB - We currently calculate area under the busulfan concentration time curve (AUC) using 7 plasma busulfan concentrations (AUC7) drawn after the first of 16 i.v. busulfan doses given as a 2-hour infusion every 6 hours. The aim of this study was to develop and validate limited sampling strategies (LSSs) using 3 or fewer busulfan concentration values with which to reliably calculate AUC in children undergoing hematopoietic stem cell transplant (HSCT). Children in the development group (44) received i.v. busulfan at Sick Kids; the validation group consisted of 35 children who received care at CHU Ste-Justine. Busulfan doses given and subsequent plasma busulfan concentrations were recorded. LSSs using 1 to 3 concentration-time points were developed using multiple linear regression. LSS were considered to be acceptable when adjusted r(2) > 0.9, mean bias <15% and precision <15%. Extent of agreement between the AUC7 values and the LSS AUC was assessed by the intraclass correlation coefficient (ICC) and Bland-Altman (BA) analysis. Agreement was considered to be excellent when the lower limit of the 95% confidence limit of the ICC exceeded 0.9 and when the limits of agreement in the BA analysis were +/-15% for both AUC and dose. Administration of the theoretic adjusted busulfan doses based on each LSS was simulated and cases where the resulting AUC was >1500 or <900 microM x min were noted. LSSs using 1, 2, or 3 plasma busulfan concentrations were developed that showed excellent agreement with AUC7 and adjusted busulfan doses. In the validation sample, only the 2- and 3-point LSSs demonstrated acceptable precision and lack of bias. LSSs using 2 or 3 plasma busulfan concentrations can be used to reliably estimate busulfan AUC after IV administration in children undergoing HSCT. PMID- 18410901 TI - Umbilical cord blood transplantation after reduced-intensity conditioning for elderly patients with hematologic diseases. AB - Although allogeneic hematopoietic stem cell transplantation is a potentially curative approach for advanced hematologic diseases, its application to elderly people is limited because of their comorbid physical conditions and lower chance of finding suitable related donors. Umbilical cord blood transplantation with reduced-intensity pretransplant conditioning (RI-UCBT) is 1 way to avoid these obstacles. We analyzed elderly patients aged 55 years and older with hematologic diseases who underwent RI-UCBT at our institute to assess feasibility and effectiveness of this treatment approach. Among the 70 patients included, 50 died, 74% of them from nonrelapse causes. Infection was the primary cause of death. Estimated overall survival and progression-free survival at 2 years were both 23%. In multivariate analyses, standard-risk diseases, age younger than 61 years, grade 0-II acute graft-versus-host disease, and the absence of preengraftment immune reaction were significantly associated with better overall survival. RI-UCBT is a potentially curative and applicable approach for elderly patients. Higher mortality, especially from nonrelapse causes, is the biggest problem to be solved to increase the feasibility of this approach. PMID- 18410902 TI - Myeloablative intravenous busulfan/fludarabine conditioning does not facilitate reliable engraftment of dual umbilical cord blood grafts in adult recipients. AB - The efficacy of once-daily intravenous busulfan with fludarabine as a preparative regimen for partially matched umbilical cord blood transplantation has not been formally studied. We randomized 10 adult patients with myeloid malignancies to receive either concurrent or sequential administration of intravenous busulfan 130 mg/m(2) once daily x 4 days and fludarabine 40 mg/m(2) daily x 4 days, followed by dual umbilical cord blood transplantation. The median combined cryopreserved total nucleated cell dose was 3.6 x 10(7)/kg recipient body weight (range: 2.8-4.5 x 10(7)/kg). Graft-versus-host disease (GVHD) prophylaxis was provided by tacrolimus and mycophenolate mofetil (MMF). Donor-derived neutrophil recovery was observed in only 2 of 10 patients, resulting in premature closure of the study as per graft failure stopping rules. We conclude that the myeloablative conditioning regimen of once-daily intravenous busulfan with fludarabine provides insufficient immunosuppression to allow for engraftment of partially matched, dual umbilical cord blood grafts. PMID- 18410903 TI - Infectious complications from high-dose chemotherapy and autologous stem cell transplantation for metastatic germ cell tumors. AB - High-dose chemotherapy with autologous stem cell transplantation (ASCT) is increasingly utilized in patients with relapsed and refractory germ cell tumors (GCT). Infectious complications are common after ASCT for hematologic malignancies, but their epidemiology in GCT patients has not been described. To identify infectious complications of ASCT for GCT, we conducted a retrospective study of patients treated at our institution, a tertiary-care cancer center in New York City between 1994 and 2006. Patients received ciprofloxacin prophylaxis but no routine antifungal or antiviral prophylaxis. In addition, patients were housed in shared rooms of 2 with standard precautions during hospitalizations. Overall, 107 patients with relapsed or refractory GCT were treated with 1-2 cycles of paclitaxel/ifosfamide and 1-3 cycles of high-dose carboplatin/etoposide with ASCT. Sixty (56%) of 107 patients developed 95 total infections, including 33 catheter-associated bloodstream infections. Fungal, viral, and nosocomial infections were uncommon. There were no infection-related deaths. In conclusion, serious morbidity from infection is uncommon among GCT patients receiving high dose chemotherapy with ASCT. Isolation and aggressive antifungal and antiviral prophylaxis is not warranted in these patients. PMID- 18410904 TI - Successful reduced-intensity conditioning allogeneic HSCT for HIV-related primary effusion lymphoma. PMID- 18410905 TI - An evaluation, at the 1-year stage, of a 3-year project to introduce practice education facilitators to NHS Tayside and Fife. AB - As a result of the Scottish Executive Health Department (SEHD) initiative, Facing the Future [Scottish Executive Health Department, 2002. Facing the Future. SEHD, Edinburgh], a need was identified for a group of experienced nurses and midwives to help support students and mentors within practice settings. It was agreed to fund one hundred "G" grade posts across NHS Scotland for an initial period of three years. As a result of this initiative, in 2004 a group of fifteen Practice Education Facilitators (PEFs) were employed by NHS Tayside and NHS Fife in Scotland. Their brief was to maintain and improve learning environments within the practice setting. This paper describes an evaluation of this new role as carried out at key stages within the first year of the project. A combination of focus groups and questionnaires were used, both to analyse the perceptions of the NHS staff that PEFs would be working with and to explore the lived experience of the PEFs during their first year in post. There was a general welcome for the role across all groups. Perceptions varied on the PEF role itself, most staff groups expecting PEFs to work mainly with students. The PEFs, however, saw the main thrust of their work with mentors, their associates and other staff. PMID- 18410906 TI - Windows and mirrors: reflections of a module team teaching the arts in nurse education. AB - The five-year experience of a group of nursing lecturers teaching the expressive arts within a Scottish degree programme is outlined and discussed. The place of the arts is contextualised within curriculum developments and module content, sequencing, thematic development, mode of delivery, assessment, student evaluation and pedagogical approaches are all addressed. Relationship to practice is discussed in terms of the art of nursing, reflection, ethics and spirituality. Future developments are discussed in terms of drawing upon the wider resources of the humanities, rather than merely expressivist sources of art. The paper concludes by encouraging the teaching of the arts in nurse education to remain responsive to practice issues and to consideration of students' learning needs. PMID- 18410907 TI - Thank-you cards: reclaiming a nursing student ritual and releasing its transformative potential. AB - The giving of a "thank-you card" to the staff of a health service in which clinical experience was gained, is common practice amongst nursing students in Australia. Group reflection, or debriefing, following the clinical experience is also a common practice. As rituals in nursing, they can become routinised, taken for-granted and have little meaning or influence. This paper discusses an educational activity devised by the author that aimed to transform a relatively innocuous practice into one that had empowering potential for students, giving them voice in the health service culture and emphasizing the need for a more humanized workforce, one that actively seeks out opportunities to give each other helpful feedback so that change is ongoing. The activity drew upon narrative pedagogy, showing students how stories combined with rituals have transformative potential not only for themselves as students, but for the entire culture of nursing. The paper argues that cultural change need not be monumental for it to have enduring effect and it is within the jurisdiction of even the most junior student. The "thank-you card" ritual that students commonly use can be reclaimed and extended so that students become activists in the building of a more humanistic, supportive nursing and learning culture. PMID- 18410908 TI - Melanin-concentrating hormone (MCH) immunoreactivity in non-neuronal cells within the raphe nuclei and subventricular region of the brainstem of the cat. AB - Neurons that utilize melanin-concentrating hormone (MCH) as a neuromodulator are localized within the postero-lateral hypothalamus and zona incerta. These neurons project diffusely throughout the central nervous system and have been implicated in critical physiological processes such as energy homeostasis and sleep. In the present report, we examined the distribution of MCH immunoreactivity in the brainstem of the cat. In addition to MCH+ axons, we found MCH-immunoreactive cells that have not been previously described either in the midbrain raphe nuclei or in the periaqueductal and periventricular areas. These MCH+ cells constituted: 1. ependymal cells that lined the fourth ventricle and aqueduct, 2. ependymal cells with long basal processes that projected deeply into the subventricular (subaqueductal) parenchyma, and, 3. cells in subventricular regions and the midbrain raphe nuclei. The MCH+ cells in the midbrain raphe nuclei were closely related to neuronal processes of serotonergic neurons. Utilizing Neu-N and GFAP immunohistochemistry we determined that the preceding MCH+ cells were neither neurons nor astrocytes. However, we found that vimentin, an intermediate-filament protein that is used as a marker for tanycytes, was specifically co-localized with MCH in these cells. We conclude that MCH is present in tanycytes whose processes innervate the midbrain raphe nuclei and adjacent subependymal regions. Because tanycytes are specialized cells that transport substances from the cerebrospinal fluid (CSF) to neural parenchyma, we suggest that MCH is absorbed from the CSF by tanycytes and subsequently liberate to act upon neurons of brainstem nuclei. PMID- 18410909 TI - Proinflammatory cytokine production by cultured neonatal rat microglia after exposure to blood products. AB - Periventricular germinal matrix hemorrhage is a devastating complication of preterm birth. Inflammation appears to play a role in brain damage after premature birth and hypoxia. The effects of rat blood plasma and serum on cytokine expression by cultured rat microglial cells were investigated. We analyzed mRNA expression levels of tumor necrosis factor (TNF)-alpha, interleukin 6 and protease activated receptor-1 and -4 by quantitative RT-PCR. Protein expression for TNFalpha was done using immunocytochemistry and ELISPOT assays. Plasma and serum had dose dependent toxic effects on microglia as measured by lactate dehydrogenase release assay and activated caspase-3 immunocytochemistry. High concentrations of plasma enhanced TNFalpha mRNA expression and protein production, while high concentrations of serum enhanced IL-6 mRNA expression. This study suggests that soluble components of blood might be differentially responsible for up regulating production of the cytokines TNFalpha and IL-6 by microglia from immature rodent brain. PMID- 18410910 TI - Voltage-gated K+ channel (Kv) subunit expression of the guinea pig spiral ganglion cells studied in a newly developed cochlear free-floating preparation. AB - The spiral ganglion accommodates the cell bodies of the acoustic nerve fibres connecting the hair cells to the central nervous system. As the ionic channels containing various voltage-gated K+ channel (Kv) subunits play pivotal roles in determining the functional properties and firing behaviour of the spiral ganglion cells (SGCs), every piece of information concerning the Kv expression of the SGCs is valuable. In the present work a comprehensive immunohistochemical analysis was performed to describe the expression of 9 Kv subunits in the guinea pig cochlea on traditional wax-embedded sections as well as employing a newly developed preparation that allowed confocal analysis, reconstruction of the three dimensional appearance and precise morphological characterisation of the SGCs. Besides determining their Kv expression patterns, differences between type I and type II SGCs were sought. SGCs showed positivity for 8 out of the 9 Kv subunit specific antibodies with varying intensity and proportion of the immunopositive cells; whereas no obvious Kv3.2 positivity could be noted. Type I and type II cells demonstrated similar expression patterns for all subunits tested, with the exception of Kv1.2, whose presence was confirmed in only 50% of the type II cells. Although the present findings suggest that type I and type II cells do not differ fundamentally in the Kv subunits they possess; they also imply that SGCs may not form a homogeneous cell population, and might provide explanation of the previously noted heterogeneity of the membrane properties of the SGCs. PMID- 18410911 TI - Excitatory amino acid transporter expression by astrocytes is neuroprotective against microglial excitotoxicity. AB - Glutamate-induced excitotoxicity is considered as a major cause of neurodegenerative disease. Excitatory amino acid transporters (EAATs) on glial cells are responsible for the homeostasis of extracellular glutamate in the central nervous system which may contribute to the prevention of excitotoxic neurodegeneration. However, the differential EAAT expression in astrocytes and microglia is not fully understood. In this study, we compared the expression of EAATs in astrocytes and microglia, and we assessed the neuroprotective and neurotoxic function of astrocytes and microglia by a co-culture system. RT-PCR analyses detected that astrocytes expressed each EAAT (EAAT1-5) whereas microglia did not express EAAT4. Western blot analyses demonstrated that astrocytes express a much larger amount of membrane-localized EAATs than microglia. Astrocytes prevented excito-neurotoxicity by the reduction of exogenous glutamate whereas microglia did not. Conversely, activated microglia released an excess of glutamate that induced excitotoxic neuronal death. Astrocytes rescued neurons from microglial glutamate-induced death in a ratio-dependent manner. Inhibition of EAATs abolished glutamate uptake and the neuroprotective effect of astrocytes, but it did not alter any microglial neurotoxic or neuroprotective effects. These results revealed that astrocytic EAATs can counteract microglial glutamate induced neuronal death whereas microglial EAATs are inconsequential to neurotoxicity and neuroprotection. PMID- 18410913 TI - Pharmacology of glucocorticoids: beyond receptors. AB - Glucocorticoid hormones are important regulators of homeostasis. They are used clinically as highly effective anti-inflammatory compounds and have been prescribed for more than fifty years for a variety of conditions. They mediate their genomic actions by binding to two different intracellular receptors in target cells. The pharmacology of glucocorticoids largely depends on ligand concentration and receptor expression levels in target tissue. However, their genomic actions also critically depend on coactivators and corepressors recruitment. We discuss how various non-receptor factors affect glucocorticoid potency and efficacy with respect to their genomic effects. Differential recruitment of coregulators may account for many ligand- and cell-specific effects of glucocorticoids. This is best illustrated by the recent identification of selective glucocorticoid receptor agonists that induce distinct conformational changes to the receptors resulting in altered protein-protein interactions and consequently different regulation of gene expression. We conclude that these new molecular insights will contribute to the design of safer glucocorticoids that retain full pharmacological properties with reduced side-effects. PMID- 18410912 TI - Neuroprotective effects of neuregulin-1 on B35 neuronal cells following ischemia. AB - We previously showed that neuregulin-1 (NRG-1) protected neurons from death in vivo following focal ischemia. The goal of this study was to develop an in vitro rat ischemia model to examine the cellular and molecular mechanisms involved in the neuroprotective effects of NRG-1 on ischemia-induced neuronal death. Rat B-35 neuroblastoma cells differentiated by serum withdrawal, developed enhanced neuronal characteristics including, neurite extension and upregulation of neuronal markers of differentiation. When B35 neurons were subjected to oxygen glucose deprivation (OGD)/reoxygenation or glutamate, widespread neuronal death was seen after both treatments. Treatment with NRG-1 immediately after OGD significantly increased neuronal survival. NRG-1 administration also resulted in a significant decrease in annexin V, an early marker of apoptosis. However, the neurotoxic actions of glutamate were unaffected by NRG-1. The neuroprotective effects of NRG-1 were prevented by an inhibitor of the phosphatidylinositol-3 kinase/Akt pathway. These results provide a new model to gain insight into the mechanisms employed by NRG-1 to protect neurons from ischemic brain injury. PMID- 18410914 TI - Regulation of G protein-coupled receptor signalling: focus on the cardiovascular system and regulator of G protein signalling proteins. AB - G protein-coupled receptors (GPCRs) are involved in many biological processes. Therefore, GPCR function is tightly controlled both at receptor level and at the level of signalling components. Well-known mechanisms by which GPCR function can be regulated comprise desensitization/resensitization processes and GPCR up- and downregulation. GPCR function can also be regulated by several proteins that directly interact with the receptor and thereby modulate receptor activity. An additional mechanism by which receptor signalling is regulated involves an emerging class of proteins, the so-called regulators of G protein signalling (RGS). In this review we will describe some of these control mechanisms in more detail with some specific examples in the cardiovascular system. In addition, we will provide an overview on RGS proteins and the involvement of RGS proteins in cardiovascular function. PMID- 18410915 TI - Immune modulation and graft protection by gene therapy in kidney transplantation. AB - Kidney transplantation represents the therapy of choice for many patients with end-stage renal disease. However, the success of renal engraftment is hindered by a number of factors, the most important of which being adverse effects of systemic immunosuppressive therapy, chronic transplant dysfunction and a severe shortage of donor kidneys. Gene therapy approaches may provide valuable strategies in each of these areas. First, gene therapy holds the potential of local therapy, thus circumventing systemic side effects of chronic immunosuppression. Second, chronic transplant dysfunction may be addressed by innovative strategies to induce local immune tolerance, immune suppression and additional graft protecting mechanisms. Third, gene therapy may be instrumental in increasing the quality of the grafts by limiting ischemia-reperfusion injury, especially in non-heart beating donors, thereby expanding the donor pool. In this article, we give an overview of the current state of gene therapy in experimental models of kidney transplantation. PMID- 18410916 TI - Cells, mediators and Toll-like receptors in COPD. AB - Chronic obstructive pulmonary disease (COPD) is a global health problem. Being a progressive disease characterized by inflammation, it deteriorates pulmonary functioning. Research has focused on airway inflammation, oxidative stress, and remodelling of the airways. Macrophages, neutrophils and T cells are thought to be important key players. A number of new research topics received special attention in the last years. The combined use of inhaled corticosteroids and long acting beta(2)-adrenoceptor agonists produces better control of symptoms and lung function than that of the use of either compound alone. Furthermore, collagen breakdown products might be involved in the recruitment and activation of inflammatory cells by which the process of airway remodelling becomes self sustaining. Also, TLR (Toll-like receptor)-based signalling pathways seem to be involved in the pathogenesis of COPD. These new findings may lead to new therapeutic strategies to stop the process of inflammation and self-destruction in the airways of COPD patients. PMID- 18410917 TI - Pharmacological manipulation of memory reconsolidation: towards a novel treatment of pathogenic memories. AB - Well-consolidated memories, when retrieved, may return to a transiently fragile state, and need to be consolidated again in order to be maintained. This process has been referred to as memory reconsolidation and presumably serves to modify or strengthen memory traces. In recent years, our understanding of the neurobiological mechanisms underlying this phenomenon has increased rapidly. Here, we will briefly review some of the pharmacological evidence, stressing a crucial role for the brain's major neurotransmitter systems, such as glutamate and noradrenaline, in memory reconsolidation. Pharmacological intervention of reconsolidation processes may have clinical relevance, especially for the treatment of psychiatric disorders that are characterized by pathological memories, including post-traumatic stress disorder and addictive behaviour. PMID- 18410918 TI - Peripheral cannabinoid CB1 receptors inhibit evoked responses of nociceptive neurones in vivo. AB - We investigated the effect of peripheral administration of a selective cannabinoid CB1 receptor agonist arachidonyl-2-choroethylamide (ACEA), on evoked responses of primary afferents in vivo. Extracellular recordings were made from filaments of the saphenous nerve that responded to noxious mechanical stimulation of their receptive fields and effects of ACEA (30 and 50 microg/100 microl, i.a.) were studied. ACEA significantly inhibited evoked responses, effects that were blocked by co-administration of the cannabinoid CB1 receptor antagonist AM251 (30 microg/100 microl). These results demonstrate a cannabinoid CB1 receptor-mediated inhibition of primary afferent nociceptor excitability and provide further support for a peripheral site of action of cannabinoids. PMID- 18410919 TI - Rho kinase inhibitors: a novel therapeutical intervention in asthma? AB - In asthma, inflammatory mediators that are released in the airways by recruited inflammatory cells and by resident structural cells result in airway hyperresponsiveness caused by increased bronchoconstriction. In addition, chronic inflammation appears to drive remodelling of the airways that contributes to the development of fixed airway obstruction and airway hyperresponsiveness in chronic asthma. Airway remodelling includes several key features such as excessive deposition of extracellular matrix proteins in the airway wall (fibrosis) and increased abundance of contractile airway smooth muscle encircling the airways. Current asthma therapy fails to inhibit these features satisfactorily. This review focuses on Rho kinase as a potential drug target in asthma, as compelling evidence from animal models and ex vivo studies suggests a central role for this enzyme and its associated signalling in acute and chronic airway hyperresponsiveness. PMID- 18410920 TI - Arginine homeostasis in allergic asthma. AB - Allergic asthma is a chronic disease characterized by early and late asthmatic reactions, airway hyperresponsiveness, airway inflammation and airway remodelling. Changes in l-arginine homeostasis may contribute to all these features of asthma by decreased nitric oxide (NO) production and increased formation of peroxynitrite, polyamines and l-proline. Intracellular l-arginine levels are regulated by at least three distinct mechanisms: (i) cellular uptake by cationic amino acid (CAT) transporters, (ii) metabolism by NO-synthase (NOS) and arginase, and (iii) recycling from l-citrulline. Ex vivo studies using animal models of allergic asthma have indicated that attenuated l-arginine bioavailability to NOS causes deficiency of bronchodilating NO and increased production of procontractile peroxynitrite, which importantly contribute to allergen-induced airway hyperresponsiveness after the early and late asthmatic reaction, respectively. Decreased cellular uptake of l-arginine, due to (eosinophil-derived) polycations inhibiting CATs, as well as increased consumption by increased arginase activity are major causes of substrate limitation to NOS. Increasing substrate availability to NOS by administration of l-arginine, l-citrulline, the polycation scavenger heparin, or an arginase inhibitor alleviates allergen-induced airway hyperresponsiveness by restoring the production of bronchodilating NO. In addition, reduced l-arginine levels may contribute to the airway inflammation associated with the development of airway hyperresponsiveness, which similarly may involve decreased NO synthesis and increased peroxynitrite formation. Increased arginase activity could also contribute to airway remodelling and persistent airway hyperresponsiveness in chronic asthma via increased synthesis of l-ornithine, the precursor of polyamines and l-proline. Drugs that increase the bioavailability of l-arginine in the airways - particularly arginase inhibitors - may have therapeutic potential in allergic asthma. PMID- 18410921 TI - Mechanisms of allergy and asthma. AB - Allergies are the result of an inappropriate reaction against innocuous environmental proteins. The prevalence and severity of allergic diseases has increased dramatically during the last decade in developed countries. Allergen specific T helper (Th) cells play a pivotal role in the pathogenesis of allergic hypersensitivity reactions. These Th cells activate a complex immune reaction that triggers the release of potent mediators and enhances the recruitment of inflammatory cells, which in turn elicit an inflammatory response that leads to the clinical symptoms of allergic disease. The current therapies for allergic diseases focus primarily on control of symptoms and suppression of inflammation, without affecting the underlying cause. However, the knowledge about the pathophysiology of allergic diseases has substantially increased, offering new opportunities for therapeutic intervention. In this review, we will focus on current insights into the mechanism of allergic reactions. PMID- 18410922 TI - Aberrant gene promoter methylation in plasma cell dyscrasias. AB - The aberrant methylation of promoter CpG island is known to be a major inactivation mechanism of tumour-related genes. To determine the clinicopathological significance of gene promoter methylation in monoclonal gammopathies, we analysed the methylation status of 6 tumour suppressor genes and their association with loss of gene function. Methylation status of the genes p14, p15, p16, hMLH1, MGMT, and DAPK was determined by methylation-specific PCR in 52 cases: 30 MM, 13 MGUS, and 9 plasmacytomas, comparing them with their protein expression by immunohistochemistry, and association between methylation status, protein expression, and clinical characteristics was assessed. The methylation frequencies were 50% for p16, 17% for p15, 10% for hMLH1, 23% for MGMT and 30% for DAPK in MM samples, and 38%, 15%, 8%, and 15% for p16, p15, MGMT and DAPK respectively in MGUS samples. In plasmacytomas samples we found methylation of p16 in 55%, p15 in 22%, MGMT in 67% and DAPK in 44%. hMLH1 was unmethylated in all cases of MGUS and plasmacytomas. Immunohistochemistry showed that gene methylation was closely associated with a loss of protein expression. Our study demonstrates that methylation-mediated silencing is a frequent event in monoclonal gammopathies: 83% of MM, 46% of MGUS and 77% of plasmacytomas have at least one gene methylated, affecting different molecular pathways involved in cell cycle, DNA repair and apoptosis. This high prevalence of aberrant promoter hypermethylation suggests that monoclonal gammopathies carry a CpG island methylator phenotype, therefore the development of new DNA demethylation agents may be a potential therapeutic use in this disease. PMID- 18410923 TI - Dual-specificity phosphatases in the hypo-osmotic stress response of keratin defective epithelial cell lines. AB - Although mutations in intermediate filament proteins cause many human disorders, the detailed pathogenic mechanisms and the way these mutations affect cell metabolism are unclear. In this study, selected keratin mutations were analysed for their effect on the epidermal stress response. Expression profiles of two keratin-mutant cell lines from epidermolysis bullosa simplex patients (one severe and one mild) were compared to a control keratinocyte line before and after challenge with hypo-osmotic shock, a common physiological stress that transiently distorts cell shape. Fewer changes in gene expression were found in cells with the severely disruptive mutation (55 genes altered) than with the mild mutation (174 genes) or the wild type cells (261 genes) possibly due to stress response pre-activation in these cells. We identified 16 immediate-early genes contributing to a general cell response to hypo-osmotic shock, and 20 genes with an altered expression pattern in the mutant keratin lines only. A number of dual specificity phosphatases (MKP-1, MKP-2, MKP-3, MKP-5 and hVH3) are differentially regulated in these cells, and their downstream targets p-ERK and p-p38 are significantly up-regulated in the mutant keratin lines. Our findings strengthen the case for the expression of mutant keratin proteins inducing physiological stress, and this intrinsic stress may affect the cell responses to secondary stresses in patients' skin. PMID- 18410924 TI - What is slow axonal transport? AB - While the phenomenon of slow axonal transport is widely agreed upon, its underlying mechanism has been controversial for decades. There is now persuasive evidence that several different mechanisms could contribute to slow axonal transport. Yet proponents of different theories have been hesitant to explicitly integrate what were, at least initially, opposing models. We suggest that slow transport is a multivariate phenomenon that arises through mechanisms that minimally include: molecular motor-based transport of polymers and soluble proteins as multi-protein complexes; diffusion; and en bloc transport of the axonal framework by low velocity transport and towed growth (due to increases in body size). In addition to integrating previously described mechanisms of transport, we further suggest that only a subset of transport modes operate in a given neuron depending on the region, length, species, cell type, and developmental stage. We believe that this multivariate approach to slow axonal transport better explains its complex phenomenology: including its bi directionality; the differing velocities of transport depending on cargo, as well differing velocities due to anatomy, cell type and developmental stage. PMID- 18410925 TI - Cellular life span and the Warburg effect. AB - Enhanced glycolysis is observed in most of cancerous cells and tissues, called as the Warburg effect. Recent advance in senescent biology implicates that the metabolic shift to enhanced glycolysis would be involved in the early stage during multi-step tumorigenesis in vivo. Enhanced glycolysis is essential both in the step of immortalization and transformation, as it renders cells resistant to oxidative stress and adaptive to hypoxic condition, respectively. ES, immortalized primary, and cancerous cells display the common concerted metabolic shift, including enhanced glycolysis with reduced mitochondrial respiration by poorly characterized mechanism. Discovery of a novel regulatory mechanism for such a metabolic shift might be essential for the future development of cancer diagnosis and anti-cancer therapy. PMID- 18410927 TI - Subthalamic nucleus stimulation in Parkinson's disease: the other side of the medallion. PMID- 18410926 TI - Effects of intrathecal administration of pituitary adenylate cyclase activating polypeptide on lower urinary tract functions in rats with intact or transected spinal cords. AB - Effects of intrathecally administered pituitary adenylate cyclase activating polypeptide-38 (PACAP-38, 0.1-30 microg) on lower urinary tract function were examined in unanesthetized, decerebrate rats with an intact spinal cord and after chronic spinal cord transection (SCT). PACAP-38 was also studied in rats with intact or bilaterally transected hypogastric nerves (HGNs), to determine if sympathetic pathways to the bladder influenced responses. In SCT rats with intact HGNs under isovolumetric conditions, 30 mug of PACAP-38 but not lower doses (0.1 10 microg) increased (mean 194%) bladder contraction amplitude (BCA). In SCT rats with sectioned HGNs, 10 microg and 30 microg of PACAP-38 increased BCA by 62% and 195%, respectively. On the other hand, during continuous infusion cystometrograms (CMGs) in SCT rats with intact or sectioned HGNs, PACAP-38 (10 microg and 30 microg) markedly reduced or completely suppressed BCA (60% and 90%, respectively) and reduced external urethral sphincter (EUS) EMG activity (58% and 91%, respectively). During CMGs in spinal cord intact rats, with intact HGNs PACAP-38 30 microg increased BCA (26%) but after HGN section PACAP-38 10 microg and 30 microg increased BCA by 21% and 35%. These results suggest that after SCT, PACAP 38 activates spinal circuitry to facilitate the parasympathetic outflow to the urinary bladder and that the elimination of sympathetic pathways enhances this effect. The decrease in BCA by PACAP-38 during CMGs in SCT rats is most reasonably attributed to a reduction in urethral outlet resistance due to suppression of excitatory EUS reflexes. PMID- 18410928 TI - Selective ablation of proliferating astrocytes does not affect disease outcome in either acute or chronic models of motor neuron degeneration. AB - Astrocytes play important roles in normal CNS function; however, following traumatic injury or during neurodegeneration, astrocytes undergo changes in morphology, gene expression and cellular function known as reactive astrogliosis, a process that may also include cell proliferation. At present, the role of astrocyte proliferation is not understood in disease etiology of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), a fatal motor neuron disorder that is characterized by a relatively rapid degeneration of upper and lower motor neurons. Therefore, the role of astrocyte proliferation was assessed in both acute and chronic mouse models of motor neuron degeneration, neuroadapted sindbis virus (NSV)-infected mice and SOD1(G93A) mice, respectively. While astrocytes proliferated in the lumbar spinal cord ventral horn of both disease models, they represented only a small percentage of the dividing population in the SOD1(G93A) spinal cord. Furthermore, selective ablation of proliferating GFAP(+) astrocytes in 1) NSV-infected transgenic mice in which herpes simplex virus-thymidine kinase is expressed in GFAP(+) cells (GFAP-TK) and in 2) SOD1(G93A)xGFAP-TK mice did not affect any measures of disease outcome such as animal survival, disease onset, disease duration, hindlimb motor function or motor neuron loss. Ablation of dividing astrocytes also did not alter overall astrogliosis in either model. This was likely due to the finding that proliferation of NG2(+) glial progenitors were unaffected. These findings demonstrate that while normal astrocyte function is an important factor in the etiology of motor neuron diseases such as ALS, astrocyte proliferation itself does not play a significant role. PMID- 18410929 TI - Echinococcus multilocularis: the parasite-host interplay. AB - Alveolar echinococcosis (AE) is a severe chronic helminthic disease caused by the intrahepatic tumor-like growth of the metacestode of Echinococcus multilocularis. Metacestodes are fluid-filled, asexually proliferating vesicles, which are entirely covered by the laminated layer, an acellular carbohydrate-rich surface structure that protects the parasite from immunological and physiological reactions on part of the host. The E. multilocularis metacestode has acquired specific means of manipulating and using the immunological host response to its own advantage. These include the expression of distinct immunoregulatory parasite molecules that manipulate and interfere in the functional activity of macrophages and T cells. Recent research findings have led to a better understanding of the protein- and glycoprotein composition of the laminated layer and the E/S fraction of the metacestode, including Em2- and Em492-antigens, two metacestode antigen fractions that exhibit immunosuppressive or -modulatory properties. Understanding of the events taking place at the host-parasite interface is the key for development of novel immuno-therapeutical and/or chemotherapeutical tools. PMID- 18410930 TI - Parasitic leiomyoma in the abdominal wall after laparoscopic myomectomy. AB - OBJECTIVE: To report a case of parasitic leiomyoma of the abdominal wall after laparoscopic myomectomy. DESIGN: Case report. SETTING: Large medical center. PATIENT(S): A 31-year-old woman with a newly developed palpable mass in the abdominal wall near the trocar site of a previous laparoscopic myomectomy. INTERVENTION(S): Excision of the mass. MAIN OUTCOME MEASURE(S): Histopathologic examination of the mass. RESULT(S): A mass measuring 3.2 x 2.2 x 2.0 cm was excised. The lesion was located in the subfascial area of the abdominal wall, near the trocar site of a previous laparoscopic myomectomy. Histopathologic examination confirmed leiomyoma. CONCLUSION(S): This result shows that myoma fragments sometimes are found to be scattered in the abdominal cavity after laparoscopic myomectomy, suggesting that all myoma fragments, however small they may be, must be completely removed. PMID- 18410931 TI - Oocyte vitrification--women's emancipation set in stone. AB - The techniques of vitrification of oocytes and the subsequent warming process being used today are now producing results far superior to the results that are obtained with slow-freezing techniques, and it would seem that this is the method of female fertility preservation that will be widely used in the near future. The reported success of the use of this method should stimulate a renewed debate on oocyte storage for fertility preservation without a medical indication. PMID- 18410932 TI - GnRH antagonists may affect endometrial receptivity. AB - OBJECTIVE: HOXA10 is an essential regulator of endometrial receptivity. To determine the effect of GnRH antagonists on endometrial receptivity, we assessed endometrial HOXA10 expression in GnRH antagonist, GnRH agonist, and natural cycles. DESIGN: Prospective case-control study. SETTING: University academic medical center. PATIENT(S): Nineteen subjects were included: 12 subjects underwent controlled ovarian hyperstimulation with recombinant FSH and used either a GnRH antagonist or a GnRH agonist; seven control subjects underwent natural cycles. INTERVENTION(S): Pipelle endometrial biopsies were obtained 11 days after hCG administration or spontaneous LH surge in untreated cycles, respectively. Immunohistochemistry was used to assess HOXA10 protein expression in endometrial glands and stroma. MAIN OUTCOME MEASURE(S): Endometrial HOXA10 protein expression. RESULT(S): HOXA10 expression was significantly decreased in endometrial stromal cells in GnRH antagonist-treated cycles compared with GnRH agonist-treated cycles or natural cycle control subjects. There was no significant difference in glandular cell HOXA10 expression among the three groups. CONCLUSION(S): Use of GnRH antagonists may be associated with impaired HOXA10 expression in endometrial stromal cells and thus may affect endometrial receptivity. PMID- 18410933 TI - Characterization of natural killer cells in nonobese diabetic/severely compromised immunodeficient mice during pregnancy. AB - OBJECTIVE: To characterize uterine natural killer (uNK) cells in nonobese diabetic/severely compromised immunodeficient (NOD/SCID) mice and investigate the potential role of these cells in pregnancy tolerance. DESIGN: An animal model based study. SETTING: Academic research center in a university. ANIMAL(S): Syngeneic pregnant NOD/SCID mice were compared with non-immunodeficient BALB/c mice. INTERVENTION(S): Induction of Toll-like receptor (TLR) agonists. MAIN OUTCOME MEASURE(S): Flow cytometric analysis was performed to detect the percentage of cell subsets, and standard (51)Cr release assay was performed to determine cytotoxicity. RESULT(S): The dominant subset of uNK cells in NOD/SCID mice is DX5 (CD49b)(+), asialo ganglio-N-tetraosylceramide(+), CD25(+), CD122(+), Thy-1 (CD90)(hi), c-kit (CD117)(hi), and interleukin-10(+). In addition, the percentage of interferon-gamma(+) subset was slightly increased in response to selected TLR agonists in the NOD/SCID mice, whereas the corresponding percentage in BALB/c mice could be increased dramatically. Such an effect could be abrogated by inhibitors, including LY294002, SP600125, and PD98059. The significant increase of interferon-gamma(+) NK cell percentage in BALB/c mice was concomitant with the increase of the embryo resorption rate. In contrast, the resorption rate in NOD/SCID mice was not significantly increased upon the induction of polyinosinic polycytidylic acid or lipopolysaccharide. As expected, the NK cells from NOD/SCID mice display a detectable but lower cytotoxicity than BALB/c, as determined by standard (51)Cr release assay. In addition, the uNK cells from NOD/SCID mice also display a hyposensitivity to lipopolysaccharide-induced production of inducible nitric oxide synthase. CONCLUSION(S): A considerable percentage of immature NK cells were detected at the fetomaternal interface in NOD/SCID mice. These cells were hyposensitive to the stimulation of selected TLR agonists. Such a status seemed to be beneficial for the maintenance of pregnancy. PMID- 18410934 TI - Advantage of conservative surgical management of large ovarian neoplasms in adolescents. AB - OBJECTIVE: To measure the postoperative size and volume of the ovary after cystectomy for a neoplasm >or=10 cm. DESIGN: Retrospective case series. SETTING: Academic medical center. PATIENT(S): Nine patients who underwent surgical management for an ovarian neoplasm at Children's Hospital Boston between November 2003 and November 2007. INTERVENTION(S): Patients were evaluated by ultrasound postoperatively. MAIN OUTCOME MEASURE(S): Sonographic size and volume of the ovary after cystectomy for an ovarian neoplasm >or=10 cm. RESULT(S): The mean age of the patients was 14.2 +/- 2.2 years, ranging between 10 and 17 years. The mean preoperative size of the ovarian neoplasm was 14.8 +/- 3.8 cm, ranging between 10 and 20 cm. Serum tumor markers were negative in seven out of nine patients. There were no borderline tumors or malignancies identified. There was no statistically significant difference between the size or volume of the affected ovary and the contralateral ovary at the time of postoperative evaluation. CONCLUSION(S): Postoperative ultrasound revealed that the affected ovary resumed its normal size and volume despite the attenuated appearance of the ovarian cortex at the time of surgery. Ovarian cystectomy should be the preferred surgical approach for adolescents to ensure conservation of ovarian tissue. PMID- 18410935 TI - Sexual and organ function in patients with symptomatic prolapse: are pessaries helpful? AB - OBJECTIVE: To evaluate quality of life and pelvic organ and sexual function before and during pessary use in patients with symptomatic pelvic organ prolapse and to determine reasons which lead to cessation of pessary use. DESIGN: Prospective observational study. SETTING: Tertiary referral center. PATIENT(S): Patients with symptomatic stage II or more prolapse of the anterior, posterior, or apical vaginal wall with or without uterus were included in this study. INTERVENTION(S): We used the Female Sexual Function Index questionnaire and the Sheffield prolapse questionnaire. For quality of life we used the King's Health Questionnaire. MAIN OUTCOME MEASURE(S): Main outcome measures were quality of life and sexual and pelvic organ function. RESULT(S): A total of 73 women participated in this study; 31 were sexually active. Desire, lubrication, and sexual satisfaction showed statistically significant improvement, and orgasm remained unchanged. Statistically significant improvement in the feeling of bulge occurred during therapy, stool outlet problems were significantly improved, overactive bladder symptoms were significantly better, and pessaries did not significantly alter incontinence. CONCLUSION(S): Pessaries have been shown to be a viable noninvasive treatment for pelvic organ prolapse improving organ and sexual function as well as general wellbeing. PMID- 18410936 TI - Endometriosis of the ureter and bladder are not associated diseases. AB - OBJECTIVE: To verify whether bladder and ureter endometriosis had the same clinical features and disease behavior. DESIGN: Case-control study. SETTING: Multidisciplinary group in Sao Paulo, Brazil. PATIENT(S): A total of 690 patients were submitted to laparoscopy with histologically diagnosis of endometriosis between July 1999 and December 2006. Twelve of these patients had lesions affecting the ureter and 26 had lesions affecting the bladder. A control group consisted of 652 patients in whom endometriosis was not affecting either the ureter or the bladder. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Clinical and surgical features of patients with ureteral or bladder endometriosis. RESULT(S): No patients with ureteral endometriosis had lesions affecting the bladder. Compared with the control group, patients with ureteral endometriosis had more advanced disease (Stages III and IV) according to the American Society of Reproductive Medicine (ASRM) staging classification (100% vs. 65.5%); they also had more retrocervical (83.3% vs. 21.6%) and rectum-sigmoid lesions (91.7% vs. 17.9%). Compared with the control group, more patients with bladder endometriosis had cyclic dysuria and/or hematuria (34.6% vs. 9.8%), more advanced stages of the disease (88.4% vs. 65.5%), and an association with endometriosis of the rectum-sigmoid (65.3% vs. 17.9%). CONCLUSION(S): Ureter endometriosis is not associated with the bladder disease; however, it is associated with advanced ASRM stages and with retrocervical and rectum-sigmoid lesions. PMID- 18410937 TI - Prevention of recurrent adnexal torsion. AB - OBJECTIVE: To report a case of adnexal torsion after in vitro fertilization (IVF) with two subsequent episodes of contralateral adnexal torsion and a novel approach for reducing the risk of recurrence. DESIGN: Case report. SETTING: University-based IVF program. PATIENT(S): A 32-year-old woman who conceived with IVF and experienced sequential bilateral adnexal torsion. Left adnexal torsion was diagnosed with laparoscopic detorsion performed 2 days after embryo transfer. At 7 weeks' gestation, right adnexal torsion occurred and was managed with laparoscopic detorsion. Subsequently, right adnexal torsion recurred at 10 weeks' gestation, and laparoscopic detorsion with shortening of the uteroovarian ligament was performed. INTERVENTION(S): Gonadotropin ovulation induction, IVF, and laparoscopic detorsion of both right and left adnexa with shortening of the right uteroovarian ligament. MAIN OUTCOME MEASURE(S): Preservation of adnexa after torsion and successful pregnancy. RESULT(S): Successful pregnancy and birth; resolution of torsion, prevention of recurrence with viable bilateral adnexa after detorsion and shortening of the utero-ovarian ligament with novel use of laparoscopic Endoloop. CONCLUSION(S): This is a unique case of multiple episodes of adnexal torsion following IVF with a new form of treatment using the laparoscopic Endoloop. Management of the infertility patient should be conservative and warrants ovarian preservation whenever possible. Multiple sequential episodes of adnexal torsion during a single pregnancy are a rare complication of IVF. Shortening of the utero-ovarian ligament is an alternative to oophoropexy to prevent recurrence. PMID- 18410938 TI - Comparison of ultraminilaparotomy for myomectomy through midline vertical incision or modified Pfannenstiel incision--a prospective short-term follow-up. AB - OBJECTIVE: To evaluate the short-term therapeutic outcome of myomectomy using ultraminilaparotomy (UMLT) through a midline vertical incision (MVI) or a modified Pfannenstiel incision (MPI) in the treatment of myomas. DESIGN: Controlled, nonrandomized clinical study. SETTING: University-affiliated medical center. PATIENT(S): Ninety-eight patients with symptomatic, uncomplicated myomas warranting myomectomy. Forty-three patients underwent UMLT myomectomy by MVI and 55 by MPI. INTERVENTION(S): UMLT myomectomy through MI or MPI access. MAIN OUTCOME MEASURE(S): The outcome was measured by comparing incision length, blood loss, operative time, postoperative pain, complications, success rate, postoperative recovery, and the return to work capability in both groups. RESULT(S): General characteristics of the patients were similar in both groups. There were no statistical differences in postoperative recovery, complications, and success rate between the two groups. However, the operative technique seemed to be easier and more acceptable in the MVI group compared with that in the MPI group, because of the smaller incision wound, less operation time, and less blood loss. By contrast, less postoperative pain and an earlier return to work capability were noted in the MPI group. CONCLUSION(S): This study has demonstrated that UMLT myomectomy using either a MVI or MPI can be applied in the successful management of uncomplicated myomas. The MPI technique was more complicated, but yielded less wound pain and earlier postoperative recovery for the women during this 1-year short-term follow-up. PMID- 18410939 TI - Fibrotic tissue in the endometrioma capsule: surgical and physiopathologic considerations from histologic findings. AB - Ovarian tissue inadvertently excised along with endometriomas was associated with the presence of pericystic fibrosis and serum levels of CA-125, representing the only clinical parameter for predicting follicle loss before surgery. A consecutive series of 91 ovarian endometriotic cysts were studied, and an inverse relationship between serum CA-125 and the thickness of the fibrotic tissue of the endometrioma capsule was demonstrated. PMID- 18410940 TI - Impact of obesity on women's health. AB - OBJECTIVE: To review the impacts of obesity on women's reproductive health and fertility. DESIGN: Literature review. SETTING: Academic medical center. PATIENT(S): Forty articles are referenced. The number of patients evaluated in each of the studies varies from 33 to 213,208. INTERVENTION(S): Articles were identified from an Ovid/Medline search using the search terms obesity, dysfunctional uterine bleeding, contraception, miscarriage, infertility, and weight loss. MAIN OUTCOME MEASURE(S): The impacts of obesity on reproductive health and fertility. RESULT(S): Obesity is associated with early puberty, aberrant menstrual patterns, decreased contraceptive efficacy, ovulatory disorders, an increased miscarriage rate, and worse assisted reproductive technology outcomes. Losing weight can ameliorate many of these problems. CONCLUSION(S): Obesity is one of the most significant causes of morbidity and mortality in the U.S. Providers must educate patients about the impacts of obesity on reproductive health and fertility. PMID- 18410941 TI - A multicenter randomized comparison of the Endocapsule and the Pillcam SB. AB - BACKGROUND: Video capsule endoscopy has been shown to be the single most effective endoscopic procedure for identifying the source of obscure GI bleeding (OGIB). OBJECTIVE: Our purpose was to report on the Food and Drug Administration pivotal trial in which the Endocapsule (EC) (Olympus America, Allentown, Pa) was compared with the Pillcam SB (PSB) (Given Imaging, Yoqneam, Israel) in patients with OGIB. DESIGN: A novel trial design was used in which the EC and the PSB were swallowed by the same patient 40 minutes apart, in randomized order. SETTING: Four academic medical centers. PATIENTS: Patients with OGIB aged 18 to 85 years who had either been transfused or who had a hematocrit of <31% in males or <28% in females. INTERVENTIONS: Both video capsules were swallowed in random order. Videos from the PSB were read locally for patient management. All videos were then read by at least 2 independent readers for normal versus abnormal categorization, a diagnosis, capsule transit time, reading time, and a subjective assessment of image quality. MAIN OUTCOME MEASUREMENT: Categorization of videos as either normal or abnormal. RESULTS: Data from 51 of 63 enrolled patients were analyzed. Nine patients were excluded for technical reasons and 3 for protocol violations. Twenty-four videos were read as normal and 14 as abnormal from both capsules. Disagreement occurred in 13. No adverse events were reported for either capsule. Overall agreement was 38 of 51 (74.5%) with a kappa of 0.48, P = .008. LIMITATIONS: Although ingestion order was randomized, the videos could not be read blind owing to a different shape of the image margin. CONCLUSIONS: (1) Both devices were safe and had a comparable diagnostic yield within the range previously reported. (2) There was a subjective difference in image quality favoring the EC. (3) This study design provided unique information about capsule movement in the small intestine and the lack of electromechanical interference between 2 different capsules. PMID- 18410942 TI - A hemolytic method for the measurement of nephritic factor. AB - The absence of a simple and widely applicable test for the measurement of NF activity has hampered the accumulation of evidence bearing on its nephritogenicity. The extensive modification of a screening test for this autoantibody, reported here, has increased the range and precision of the test and made it less laborious. C3b deposited on sheep E by the reaction of NF with NHS forms a C5 convertase which, with addition of rat EDTA serum, leads to hemolysis of the cells proportionate under the right conditions to the concentration of NF in the reaction mixture. The calibration line is straight or slightly concave and passes through the origin. The method detects the activity of both the NF of the amplification loop, NFa, found in MPGN type II, and the NF of the terminal pathway, NFt, found in MPGN types I and III. Interday coefficients of variation ranged from 6.6% to 13.5% and intraday from 7.0% to 12.6%. Although serum C3 levels can be markedly depressed when NF levels are high, C3 levels and NF activity generally correlate poorly. The C3 level could be low and NF absent or, occasionally, NF present with the C3 level normal. NF activity was absent from the stored serum of patients with active SLE, AGN or with an IgA nephropathy. PMID- 18410943 TI - Heterorhabditidoides chongmingensis gen. nov., sp. nov. (Rhabditida: Rhabditidae), a novel member of the entomopathogenic nematodes. AB - During a recent soil sample survey in Eastern China, a new entomopathogenic nematode species, collected from the Chongming Islands in the southern-eastern area of Shanghai, was discovered. Morphological characteristics of different developmental stages of the nematode combined with molecular data showed that this nematode is a new genus of Rhabditidae, and described as Heterorhabditidoides chongmingensis gen. nov., sp. nov., for that it shares more morphological characteristics with heterorhabditids than with steinernematids. For males, the papillae formula of bursa is 1, 2, 3, 3, with constant papillae number in the terminal group, stoma tubular-shaped and about 1.5 head width; cheilorhabdions cuticularized, esophageal collar present and long, median bulb present. For infective juveniles, EP=90 (80-105)microm, ES=104 (92-120)microm, tail length=111 (89-159)microm, and a=19.1 (15-21). The percentages of the nucleotides A, T, C and G in the ITS1 regions of the new species are significantly different from those of heterorhabditids and other rhabditids. Molecular phylogenetic trees based on 18S rDNA and the internal transcribed spacer (ITS) sequences data revealed that the new entomopathogenic nematode species forms a monophyletic group, which is a sister group of the clade comprised of some genera of Rhabditidae. PMID- 18410944 TI - Functional role of Notch signaling in the developing and postnatal heart. AB - In the developing heart, Notch signaling plays an essential role in several key developmental processes, such as epithelial-to-mesenchymal transition and myocyte proliferation and differentiation. The importance of Notch in cardiac development has been demonstrated in knockout mice carrying null mutations in genes encoding components of the Notch pathway. Furthermore, humans with inactivating mutations in the Notch ligand Jagged1 suffer from Alagille syndrome, a condition characterized by several cardiac defects. Notch1 receptor haploinsufficiency has also been involved in aortic valve disease in humans. In addition, accumulating evidence indicates that Notch may also regulate homeostasis in the adult heart. Notch may protect the heart from an excessive and detrimental hypertrophic response and increase cardiomyocyte survival. Emerging evidence also suggests that Notch could be important for cardiac tissue renewal by controlling the maintenance and commitment of a cardiac stem cell compartment. PMID- 18410945 TI - Epidemiology of familial amyloid polyneuropathy in Japan: Identification of a novel endemic focus. AB - BACKGROUND: Familial amyloid polyneuropathy (FAP) is distributed worldwide with several endemic foci including two major foci in Japan. OBJECTIVE: To elucidate a nationwide epidemiology of FAP in Japan. DESIGN, SETTING, AND PATIENTS: (i) We analyzed the data of FAP patients registered by the Ministry of Health, Labour, and Welfare, Japan, during 2003-2005. (ii) As Ishikawa prefecture was found to be a novel endemic focus, we examined 27 FAP patients in Ishikawa to characterize their clinical and genetic features in comparison with other endemic foci. RESULTS: (i) The prevalence of familial amyloidosis in Japan was estimated to be 0.87-1.1 per 1,000,000 persons. Nagano prefecture had the highest prevalence (11 15.5), followed by Kumamoto (10.1-10.3), and then Ishikawa (3.5-4.2). (ii) All the FAP patients in Ishikawa had transthyretin (TTR) type FAP; all the families had a TTR Val30Met mutation except one family with a Leu58Arg mutation. FAP with Val30Met mutation in Ishikawa was characterized by late onset, high penetrance, and moderate autonomic dysfunction. CONCLUSIONS: Ishikawa prefecture is the third endemic focus of FAP in Japan. FAP with TTR Val30Met mutation in Japan can be classified to (i) early-onset and endemic (Nagano and Kumamoto), (ii) late-onset and endemic (Ishikawa), and (iii) late-onset and non-endemic types. PMID- 18410946 TI - Down-regulation of NR2B receptors partially contributes to analgesic effects of Gentiopicroside in persistent inflammatory pain. AB - Gentiopicroside is one of the secoiridoid compound isolated from Gentiana lutea. It exhibits analgesic activities in the mice. The anterior cingulate cortex (ACC) is a forebrain structure known for its roles in pain transmission and modulation. Painful stimuli potentiate the prefrontal synaptic transmission and induce glutamate NMDA NR2B receptor expression in the ACC. But little is known about Gentiopicroside on the persistent inflammatory pain and chronic pain-induced synaptic transmission changes in the ACC. The present study was undertaken to investigate its analgesic activities and central synaptic modulation to the peripheral painful inflammation. Gentiopicroside produced significant analgesic effects against persistent inflammatory pain stimuli in mice. Systemic administration of Gentiopicroside significantly reversed NR2B over-expression during the chronic phases of persistent inflammation caused by hind-paw administration of complete Freunds adjuvant (CFA) in mice. Whole-cell patch clamp recordings revealed that Gentiopicroside significantly reduced NR2B receptors mediated postsynaptic currents in the ACC. Our findings provide strong evidence that analgesic effects of Gentiopicroside involve down-regulation of NR2B receptors in the ACC to persistent inflammatory pain. PMID- 18410947 TI - Effects of the selective neurotensin antagonist SR 142948A on 3,4 methylenedioxymethamphetamine-induced behaviours in mice. AB - Neurotensin is one of the genes previously found up-regulated in mice striatum after acute injection of MDMA (9 mg/kg). In order to examine the pharmacological significance of this effect, the involvement of the neurotensinergic system in MDMA-induced behaviors was explored in mice using the neurotensin receptor antagonist SR142948A (1mg/kg). We found that acute administration of the antagonist inhibited the MDMA-elicited locomotor activity. SR142948A pre treatment had no effect on the acquisition of conditioned place preference (CPP) to MDMA but abolished the expression of this behavior. We also studied the effects of acute and repeated exposure to MDMA on the mRNA level of neurotensin in mice striatum. Kinetic analysis of the regulation 1, 2, 6 and 12h after acute injection of MDMA showed that the drug transiently up-regulates neurotensin mRNA in this structure. The time course of the modulation suggests that the effects observed with SR142948A are attributable to the release of a preexisting endogenous pool rather than the newly synthesized peptide. Repeated exposure to MDMA following the same injection pattern used in the CPP paradigm revealed an increase in mRNA level of neurotensin in mice striatum. These results indicate that endogenous neurotensin plays a role in both the acute locomotor activity and the expression of CPP induced by MDMA. PMID- 18410948 TI - Additive neuroprotection of GABA A and GABA B receptor agonists in cerebral ischemic injury via PI-3K/Akt pathway inhibiting the ASK1-JNK cascade. AB - Co-activation of GABA A and GABA B receptors results in neuroprotection during in vitro ischemia. However, it is unclear whether this mode of action is responsible for its neuroprotective effects in animal models of ischemia in vivo, and the precise mechanisms are also unknown. This study compared the neuroprotective efficacies of muscimol, a GABA A receptor agonist, and a GABA B receptor agonist baclofen in rat brain ischemia. The additive neuroprotection could be obtained in the hippocampal CA1 pyramidal cells prominently when muscimol and baclofen were co-applied. In particular, our study showed that co-activation of GABA A and GABA B receptors could strongly increase Akt activation and inhibit ASK1 activation by phosphorylation of serine 83 of ASK1. PI-3K inhibitor LY294002 reversed the increasing Akt activation and ASK1 (S83) phosphorylation. Moreover, MKK4/MKK7-JNK signaling activation was inhibited during ischemia/reperfusion (I/R) by co treatment of muscimol with baclofen. JNK substrate, Bcl-2 and c-jun phosphorylation were also attenuated. Our results indicated that co-activation of GABA A receptor and GABA B receptor exerted neuroprotective effect via PI-3K/Akt pathway, which could inhibit the ASK1-c-Jun N-terminal protein kinase (JNK) cascade. PMID- 18410950 TI - A spectroscopic study of the bromination of the endocrine disruptor ethynylestradiol. AB - Bromination of ethynylestradiol (EE2) was studied using fluorescence and conventional and stopped-flow absorbance spectroscopy. EE2 transformations in the presence of bromine were determined to proceed through sequential reactions. The first step is very rapid and results in the formation of a brominated cyclohexadienone that is susceptible to reduction but in oxidative conditions it undergoes a transformation into monobromo-EE2 in a first-order OH- -accelerated reaction. The transformation from monobromo-EE2 to dibromo-EE2 follows a similar mechanism. The rates of reactions of bromine-substituted EE2 species are orders of magnitude lower than that of the parent compound indicating that these species can persist in conditions typical for water treatment operations. PMID- 18410949 TI - Replication-defective virus vaccine-induced protection of mice from genital herpes simplex virus 2 requires CD4 T cells. AB - Replication-defective herpes simplex virus 2 (HSV-2), used as an immunization strategy, protects against HSV-2 challenge in animal models. The roles of replication-defective virus-induced T cell subsets in control of HSV-2 infection have not been established. Mice lacking B cells (microMT) were immunized, depleted of CD4 or CD8 T cells, and then challenged intravaginally with HSV-2 to elucidate T cell subset contributions in the absence of virus-specific antibody. Immunized, CD4-depleted microMT mice developed severe infection of the genital tract and nervous system. In contrast, depletion of CD8 T cells from microMT mice did not attenuate protection. Immunized wild-type mice depleted of CD4 T cells also developed more severe HSV-2 infection than mice from which CD8 T cells were depleted. Thus, immunization with replication-defective virus induces T cell responses that effectively control HSV-2 infection in the absence of HSV-immune antibody, and CD4 T cells play the predominant role in this protective effect. PMID- 18410951 TI - "Ni-struvite" - a new biomineral formed by a nickel resistant Streptomyces acidiscabies. AB - Biomineralization dependent on bacterial activity has been described for struvite which is formed in soils, guano, putrescent matter and sediments. A new biomineral containing nickel instead of magnesium, Ni(NH4)(PO4) . 6H2O, has been identified. It was formed by nickel resistant Streptomyces acidiscabies E13, and putatively named nickel struvite. The mineral formation is dependent on biological activity since non-viable bacterial cells are not capable to induce formation of Ni-struvite under identical conditions. Formation of Ni-struvite was observed on colony surfaces upon prolonged incubation of solid minimal or complex media containing elevated concentrations of 8-15mM NiCl2. The formation of magnesium containing crystals was not observed although Mg2+ is present in the medium. However, the process was not depending on desiccation since small crystals attached to the mycelial biomass of the bacteria were observed microscopically also in liquid cultures of nickel supplemented minimal and complex media after two weeks of incubation. The capacity to induce biomineralization of a nickel containing mineral is postulated to constitute a resistance factor, allowing the soil bacterium to withstand high nickel concentrations. The strain shows nickel resistance as an adaption to its habitat, since this bacterium was isolated from a former uranium mining site in Eastern Thuringia, Germany, where nickel concentrations of up to 2000ppm (translating to appr. 30mM) occur as a result of former mining activities. PMID- 18410952 TI - Haemostatic effects of phytoestrogen genistein in postmenopausal women. AB - INTRODUCTION: Genistein is an isoflavone phytoestrogen derived from the soybean which acts as natural selective estrogen receptor modulator. Various studies have pointed out its cardioprotective role. The aim of the study was to evaluate the haemostatic effects of genistein in postmenopausal women. MATERIAL AND METHODS: In this double-blind placebo-controlled trial we enrolled 104 healthy postmenopausal women with osteopenia. 53 patients (mean age 54.9+/-4.2 yr; BMI 23.4+/-3.2 Kg/m(2)) received genistein (54 mg/day) and 51 patients (mean age 55.4+/-4.3 yr; BMI 23.6+/-3.6 Kg/m(2)) received an identical placebo-tablet. Both groups received a calcium and vitamin D supplement. Plasma levels of D-dimer (DD), plasminogen activator inhibitor-1 (PAI-1) and prothrombin fragment 1+2 (F1+2) were measured at baseline and after 6 and 12 months of treatment. RESULTS: Baseline characteristics of the two groups were similar. Compared with placebo, genistein decreased significantly DD (p<0.001), but did not affect PAI-1 and F 1+2 plasma levels. CONCLUSION: The results of our study do not confirm effects of genistein on activation of the haemostatic system, but on the contrary the significant decrease of DD could indicate a possible cardioprotective role of genistein in postmenopausal women. PMID- 18410954 TI - Topotecan weekly bolus chemotherapy for relapsed platinum-sensitive ovarian and peritoneal cancers. AB - OBJECTIVE: Topotecan at a dose of 1.5 mg/m(2) on days 1 to 5 of a 21-day cycle is an approved therapy for recurrent ovarian cancer. However, heavily pretreated patients may be predisposed to hematologic adverse events. This prospective study, therefore, investigates the safety and efficacy of an alternate weekly schedule of topotecan in patients with recurrent ovarian or peritoneal cancer. METHODS: Patients with potentially platinum-sensitive recurrent ovarian or peritoneal cancer were treated with 4.0 mg/m(2) weekly topotecan as tolerated until disease progression. Antitumor response and safety were assessed. Dose reductions, delays, or omissions were implemented for grades 3-4 adverse events. RESULTS: Of the 41 enrolled patients (median age, 62 years; range, 42 to 82 years), 39 patients had ovarian cancer, and 2 patients had peritoneal cancer. The median platinum-free interval was 11.7 months. A median of 9 topotecan cycles (range, 1 to 45 doses) was administered. Weekly topotecan was well tolerated: 7 (17%) patients had grades 3-4 neutropenia, and 9 (22%) had grades 3-4 fatigue. No grade 4 thrombocytopenia or anemia was reported. Of 38 response-evaluable patients, 1 (3%) had a complete response, 8 (21%) had a partial response, 16 (42%) had stable disease, and 13 (34%) had progressive disease. CONCLUSIONS: Weekly topotecan was well tolerated in patients with platinum-sensitive ovarian or peritoneal cancer at first relapse, with a hematologic profile that compared favorably with that of the 5-day topotecan regimen. Moreover, antitumor activity was similar to that reported for the 5-day regimen. PMID- 18410955 TI - Changes in amygdala neural activity that occur with the extinction of context dependent conditioned fear stress. AB - The purpose of the present study was to characterize functional changes in the amygdala that accompany the extinction of context-dependent conditioned fear stress in a rat, an animal model of anxiety. Specifically, the effect of extinction of conditioned fear-induced cyclic AMP responsive element-binding protein (CREB) phosphorylation in the amygdala was investigated using immunohistochemistry. Experiments demonstrated that CREB phosphorylation in the basal nucleus of the amygdala decreased with the extinction of context-dependent conditioned fear-induced freezing behavior. These data suggest that the basal nucleus of the amygdala plays an essential role in the expression of context dependent conditioned fear. Further, this is the first study to demonstrate that CREB phosphorylation in the basal nucleus of the amygdala changes in parallel with the extinction of context-dependent conditioned fear. PMID- 18410958 TI - Evidence of a role for B cell-activating factor of the TNF family in the pathogenesis of chronic rhinosinusitis with nasal polyps. AB - BACKGROUND: The polypoid form of chronic rhinosinusitis (chronic rhinosinusitis with nasal polyps [CRSwNP]) is a highly prevalent disease that often requires surgical intervention for treatment. Nasal polyps contain large quantities of B lymphocytes and immunoglobulin as well as eosinophils. OBJECTIVES: The objective of this study was to investigate the expression of B cell-activating factor of the TNF family (BAFF), an important regulator of class-switch recombination and immunoglobulin production, in patients with chronic rhinosinusitis (CRS). METHODS: We collected nasal tissue and nasal lavage fluid from patients with CRS and control subjects. We assayed mRNA for BAFF and B-lymphocyte markers, CD20 and transmembrane activator and calcium-modulator and cyclophilin ligand interactor, by using real-time PCR, and assayed BAFF protein by using ELISA and immunohistochemistry. RESULTS: BAFF mRNA was significantly increased in nasal polyps from patients with CRSwNP (P < .001) compared with inferior turbinate tissue from patients with CRS or healthy subjects. BAFF protein was also elevated in polypoid tissue and nasal lavage from patients with CRSwNP. Immunohistochemistry showed considerable BAFF staining in mucosal epithelial cells in nasal polyps along with unidentified cells in the lamina propria. Expression of mRNA for BAFF in sinonasal tissue was significantly correlated with CD20 and transmembrane activator and CAML interactor in sinus tissue. IgA, an immunoglobulin isotype known to activate eosinophils, was also significantly elevated in the polypoid tissue. CONCLUSION: Overproduction of BAFF in nasal polyps may contribute to the pathogenesis of CRSwNP via the local induction of IgA and activation of eosinophils. PMID- 18410959 TI - Oral tolerance, food allergy, and immunotherapy: implications for future treatment. AB - The lumen of the gastrointestinal tract is exposed daily to an array of dietary proteins. The vast majority of proteins are tolerated through suppression of cellular or humoral responses, a process known as oral tolerance. However, in approximately 6% of children and 4% of adults in the United States, tolerance to a given dietary antigen either is not established or breaks down, resulting in food hypersensitivity. Although food allergies can result in sudden and life threatening symptoms, their prevalence is remarkably low considering the complexities of the gut-associated mucosal system. Suppression involves signaling by an array of nonprofessional antigen-presenting cells, dendritic cells, and regulatory T cells, as well as lymphocyte anergy or deletion. Several factors, including antigen properties, route of exposure, and genetics and age of the host, contribute to the development of oral tolerance. Although the current standard of care for patients with food allergies is based on avoidance of the trigger, increased understanding of the mechanisms involved in tolerance has shifted focus of treatment and prevention toward inducing tolerance. Data from early-phase clinical trials suggest both sublingual and oral immunotherapy are effective in reducing sensitivity to allergens. In this article we review the mechanisms of tolerance, discuss aberrations in oral tolerance, and provide information on novel prevention and treatment paradigms for food allergy. PMID- 18410961 TI - Asthma exacerbations during the first trimester of pregnancy and the risk of congenital malformations among asthmatic women. AB - BACKGROUND: Uncontrolled maternal asthma during pregnancy has been hypothesized as a cause of congenital malformation, but literature is scare on this topic. OBJECTIVE: The aim of this study was to investigate whether asthmatic women who had an exacerbation during the first trimester of pregnancy were more at risk of having a baby with a congenital malformation. METHODS: From the linkage of 3 Canadian administrative databases, we reconstructed a cohort of 4344 pregnancies of asthmatic women. Asthma exacerbations were assessed during the first trimester of pregnancy and were defined as a filled prescription for oral corticosteroids, an emergency department visit, or a hospitalization for asthma. Congenital malformations were assessed at birth and during the first year of life of the newborn by using diagnoses recorded in the databases. Generalized estimating equation models were used to estimate adjusted odds ratios of congenital malformations in association with asthma exacerbations. RESULTS: In the cohort we identified 398 (9.2%) babies with at least 1 malformation and 261 (6.0%) with a major malformation. The crude prevalences of malformations were 12.8% and 8.9%, respectively, for women who had and those who did not have an exacerbation. The adjusted odds ratio for all malformations was 1.48 (95% CI, 1.04-2.09) when comparing women who had and those who did not have an exacerbation. The corresponding figures were 1.32 (95% CI, 0.86-2.04) for major malformations. CONCLUSION: Asthma exacerbations during the first trimester of pregnancy were found to significantly increase the risk of a congenital malformation. PMID- 18410960 TI - Anti-IL-5 (mepolizumab) therapy reduces eosinophil activation ex vivo and increases IL-5 and IL-5 receptor levels. AB - BACKGROUND: Anti-IL-5 might be a useful therapeutic agent for eosinophilic disorders, yet its immunologic consequences have not been well characterized. OBJECTIVE: We sought to characterize the hematologic and immunologic effects of anti-IL-5 in human subjects. METHODS: The effects of 3-month infusions of mepolizumab were assessed in 25 patients with a variety of eosinophilic syndromes. Samples with increased IL-5 levels after therapy were analyzed by using size exclusion filtration. Immunoreactive IL-5 fraction and plasma samples were subsequently precipitated with saturating concentrations of protein A/G. RESULTS: Twenty-three patients responded to anti-IL-5 therapy with a decrease in blood eosinophil counts and a reduced percentage of CCR3(+) cells by 20- and 13 fold, respectively (P < .0001). Responsiveness was not related to the levels of baseline plasma IL-5 or the presence of FIP1L1-PDGFRA fusion gene. Persistently decreased blood eosinophilia remained for 3 months after final infusion in 76% of subjects. Therapy was associated with a large increase in blood IL-5 levels, likely because of a circulating IL-5/mepolizumab complex precipitated with protein A/G, a significant increase in eosinophil IL-5 receptor alpha expression, and increased percentage of CD4(+) and CD8(+) cells producing intracellular IL-5 (P < .05). Additionally, anti-IL-5 therapy decreased eotaxin-stimulated eosinophil shape change ex vivo. CONCLUSIONS: Anti-IL-5 therapy induces a dramatic and sustained decrease in blood eosinophilia (including CCR3(+) cells), decreased eosinophil activation, and increased circulating levels of IL-5 in a variety of eosinophilic disorders. Increased levels of IL-5 receptor alpha and lymphocyte IL-5 production after anti-IL-5 therapy suggest an endogenous IL-5 autoregulatory pathway. PMID- 18410962 TI - Anti-CD20 (rituximab) treatment for atopic eczema. PMID- 18410963 TI - The soluble form of a disintegrin and metalloprotease 33 promotes angiogenesis: implications for airway remodeling in asthma. AB - BACKGROUND: A disintegrin and metalloprotease (ADAM)-33 is a susceptibility gene for asthma and chronic obstructive pulmonary disease whose function remains unknown. OBJECTIVE: Because asthmatic bronchoalveolar lavage fluid contains high levels of soluble ADAM33 (sADAM33), which includes the catalytic domain, we postulated that its release from cell membranes might play functional roles in airway remodeling by promoting angiogenesis. METHODS: The proangiogenic activity of the highly purified catalytic domain of ADAM33 or a catalytically inactive mutant was studied in vitro (Matrigel assay), ex vivo (human embryonic/fetal lung explants) and in vivo (chorioallantoic membrane assay). The regulation of sADAM33 release from cells overexpressing full-length ADAM33 and its biological activity were characterized. RESULTS: We show that the purified catalytic domain of ADAM33, but not its inactive mutant, causes rapid induction of endothelial cell differentiation in vitro, and neovascularization ex vivo and in vivo. We also show that TGF-beta(2) enhances sADAM33 release from cells overexpressing full length ADAM33 and that this truncated form is biologically active. CONCLUSION: The discovery that sADAM33 promotes angiogenesis defines it as a tissue remodeling gene with potential to affect airflow obstruction and lung function independently of inflammation. As TGF-beta(2) enhances sADAM33 release, environmental factors that cause epithelial damage may synergize with ADAM33 in asthma pathogenesis, resulting in a disease-related gain of function. This highlights the potential for interplay between genetic and environmental factors in this complex disease. PMID- 18410965 TI - Nonylphenol and octylphenol in human breast milk. AB - Human milk is the most important form of nourishment for newborn children. Its consumption is strongly recommended by health authorities also for other important advantages. Unfortunately, in the last three decades a great number of investigations have shown the occurrence of several environmental contaminants in human milk, especially those with lipophilic properties. This study investigates the presence of nonylphenol, octylphenol (OP), nonylphenol monoethoxylate (NP1EO) and two octylphenol ethoxylates (OPEOs) (namely OP1EO and OP2EO), in human breast milk of Italian women. NP was the contaminant found at the highest levels with mean concentrations of 32 ng/mL, about two orders of magnitude higher than OP (0.08 ng/mL), OP1EO (0.07 ng/mL) and OP2EO (0.16 ng/mL). In the group of study a positive correlation among fish consumption and levels of NP in the milk was observed, in accordance with the evidence that seafood represents one of the most important sources of exposure to this group of contaminants in Italy. On the basis of the concentrations found in the breast milk samples, a maximum NP daily intake of 3.94 microg/kg/day can be calculated, which is close to the Tolerable Daily Intake (TDI) of 5 microg/kg body weight (bw) proposed by the Danish Institute of Safety and Toxicology. In the cases of OP no TDI is available, but its intake is at least six orders of magnitude lower than the NOAEL of 10 mg/kg/day derived from a two generation study on rats. PMID- 18410964 TI - The CCK-system underpins novelty-seeking behavior in the rat: gene expression and pharmacological analyses. AB - Cholecystokinin (CCK) and its receptor CCK-2R have been shown to promote emotional responsivity and behavioral sensitization to psychostimulants in the rat. An animal model has been developed based on locomotor response to a novel inescapable environment. Animals exhibiting consistent differences in locomotor response to novelty have been termed as high and low responder rats (HR and LR, respectively). This paradigm is deemed to model sensation-seeking, a personality trait closely associated with substance abuse. The present study provides genetic and pharmacological evidence that the CCK-ergic system modulates this behavior. Distinctive patterns of CCK-related gene expression in HR and LR animals occurred beyond the mesolimbic pathways. CCK gene expression was higher in hippocampus, amygdala, and prefrontal cortex, but lower in the ventral tegmental area of HR relative to LR rats. Levels of CCK-2R mRNA were more elevated in LR animals in some areas of the forebrain such as the prefrontal cortex, nucleus accumbens, and hippocampus. Additionally, CCK-2R blockade with the antagonist LY225.910 (0.5 mg/kg) removed phenotype differences in sustained exploration of novel stimuli (i.e., a novel-object) in HR and LR rats exposed to an enriched open-field test series. Finally, CCK-2R blockade also altered M(2) and 5-HT(7) receptor gene expression in the mediodorsal thalamus (a strategic structure for corticothalamic trafficking) in a phenotype-dependent manner. Taken together, the findings reported here suggest that distinct CCK-ergic function may contribute to promoting individual differences in novelty-seeking behavior. PMID- 18410966 TI - Mutagenic properties of PM2.5 air pollution in the Padana Plain (Italy) before and in the course of XX Winter Olympic Games of "Torino 2006". AB - PM2.5 is one of the most important aspects of environmental health. This air pollutant is breathable and it is implicated in several chronic adverse health effects such as the decrease of respiratory functionality and cancer. Several in vitro bioassays are able to predict the mutagenic/carcinogenic activity of the environmental pollutants and mixtures of them. In this study PM2.5 air pollution was daily monitored in three cities located in the Northern part of Italy and the mutagenic properties of the PM2.5 organic extracts were also assessed. Samplings lasted 14 months and cover the period of the Winter Olympic Games of "Torino 2006". In this work, the levels of PM2.5, its mutagenic properties (detected with Salmonella typhimurium assay), the role of the Olympic Games as environmental factor and some meteorological data are discussed. The mean concentration of PM2.5 measured in Torino was 45.4 (+/-30.6) microg/m3, in Pavia 37.6 (+/-25.6) microg/m3, in Verona 43.1 (+/-28.5) microg/m3. Findings of the monthly pool bioassay were in Torino 107 (+/-104) net revertans/m3, in Pavia 108 (+/-89) net revertans/m3, in Verona 128 (+/-109) net revertans/m3. The Olympic Games period data show that PM2.5 pollution and its load of mutagenic potential are different and partially independent phenomena. The Olympic Games had not a great impact on the PM2.5 pollution. The exclusive PM2.5 gravimetric analysis shows a potential human risk if compared with the latest international guide values but it does not describe exhaustively the human health risk associated to the presence of this particular air pollutant. Moreover, the chemical and biological activity qualification of the PM organic extracts as a whole, can instead improve the knowledge. PMID- 18410967 TI - The clinical diagnosis of Legionnaires' disease: the diagnostic value of combining non-specific laboratory tests. PMID- 18410968 TI - Treatment of Clostridium difficile disease in patients not responding to metronidazole. PMID- 18410969 TI - A prospective evaluation of the Infection Probability Score (IPS) in the intensive care unit. AB - OBJECTIVES: Identification of infection remains a major challenge, particularly in acutely ill patients. The Infection Probability Score (IPS) was developed to help rule out infection in acutely ill patients. In the present study, we determined the IPS in acutely ill, intensive care unit (ICU) patients to assess its use in the diagnosis and treatment of infection. METHODS: In this prospective, observational study, we enrolled 107 consecutive patients who were admitted to the ICU without antibiotic therapy. Patients were allocated to four groups according to the probability of infection determined from clinical and microbiological data and their IPS values were then evaluated daily throughout the ICU stay. RESULTS: The IPS was higher in patients with the highest clinical probability of infection and decreased significantly in these patients after 5 days of effective antimicrobial therapy. The IPS remained below the cut-off value in non-infected patients. Patients in whom inadequate antimicrobial therapy was administered had a greater mortality than the other patients. CONCLUSIONS: The IPS had a good predictive value for diagnosis of infection. In addition, dynamic evaluation of this score may help to assess the response to therapy. PMID- 18410970 TI - Interventions that are helpful for depression and anxiety in young people: a comparison of clinicians' beliefs with those of youth and their parents. AB - BACKGROUND: The aim of the research was to assess the consensus of clinicians about the appropriate treatments for depression and anxiety in youth, to discover any major differences between clinicians' beliefs and those of young people and their parents, and to assess whether clinicians' beliefs were consistent with clinical practice guidelines for young people. METHODS: Postal surveys were carried out on the beliefs of Australian general practitioners, psychiatrists, psychologists, and mental health nurses about what are helpful interventions for depression and social phobia in a 15-year old and in a 21-year old. The clinician data were compared to findings from an earlier national survey of Australian youth aged 12-25 years and their parents. RESULTS: The clinicians showed consensus about the helpfulness of a number of professions, reducing substance use, cognitive-behavior therapy, counseling, physical activity, relaxation training, and (for social phobia) meditation. Antidepressants were generally recommended only for depression in a 21-year old. Young people and their parents showed much lower endorsement than clinicians of antidepressants and cognitive behavior therapy, while clinicians had much lower endorsement than the public of informal supports such as family, friends and support groups. LIMITATIONS: The survey was adapted from one designed for the public and did not allow clinicians to express complex opinions. CONCLUSIONS: Clinicians show substantial consensus about appropriate treatments. However, there are some major differences in beliefs between clinicians and young people, which may be a barrier to optimal help-seeking. PMID- 18410971 TI - Soluble tumor necrosis factor receptor 1 and tissue inhibitor of metalloproteinase-1 in hemolytic uremic syndrome with encephalopathy. AB - Enterohemorrhagic Escherichia coli (EHEC) induces hemorrhagic colitis and hemolytic uremic syndrome (HUS). Morbidity and mortality are increased in HUS patients with neurologic complications. To determine the pathogenesis of the central nervous system (CNS) involvement in HUS by EHEC, we determined the serum concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), soluble TNF receptor 1 (sTNFR1), IL-10, interferon-gamma (IFN-gamma), IL-2, IL-4, soluble E-selectin (sE-selectin), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1) during the acute stage in children with HUS with or without CNS involvement. Serum concentrations of IL-6, IL-10, sTNFR1, sE-selectin, MMP-9, and TIMP-1, but not TNF-alpha, IFN-gamma, IL-2, or IL-4, were significantly higher in patients with HUS with encephalopathy compared with controls. Serum IL-6, sTNFR1 and TIMP-1 concentrations were significantly higher in patients with HUS with encephalopathy compared with those with HUS without encephalopathy (P=0.031, P=0.005, and P=0.007, respectively) and those with acute colitis without HUS (P=0.011, P<0.001, and P=0.005, respectively). There were no significant differences in hemoglobin, platelet counts, leukocyte counts, or serum concentrations of IL-10, sE-selectin, MMP-9, aspartate aminotransferase, lactate dehydrogenase, blood urea nitrogen, creatinine, or C-reactive protein between the HUS patients with and without encephalopathy. Our preliminary study suggests that serum IL-6, sTNFR1 and TIMP-1 levels, particularly sTNFR1 and TIMP 1, are important for predicting neurological complications in patients with HUS. PMID- 18410972 TI - Brain Natriuretic Peptide (BNP) regulates the production of inflammatory mediators in human THP-1 macrophages. AB - Brain Natriuretic Peptide (BNP), besides retaining vasodilatory, diuretic and natriuretic properties, is a vasoactive hormone that it is also involved in several cardiac diseases as well as severe sepsis and septic shock. All these conditions are characterized by an ongoing inflammatory response consisting in a complex interaction of pleiotropic mediators derived from plasma or cells, including monocytes and macrophages. However, the relationship between this hormone and inflammation remains to be elucidated. Therefore, the aim of the present study was to evaluate a possible BNP immunomodulatory activity on macrophages. Our results demonstrate that BNP regulates the production of major inflammatory molecules, such as reactive oxygen- and nitrogen species (ROS and RNS), leukotriene B(4) (LTB(4)), prostaglandin E(2) (PGE(2)); modulates the cytokines (TNF-alpha, IL-12 and IL-10) profile, and affects cell motility. These results furnish novel and brand-new proofs on BNP ability of modulating the production of inflammatory mediators in macrophages whose role has broad implications in inflammatory states where increased BNP levels have been reported. PMID- 18410973 TI - Catalyzed reporter deposition-fluorescent in situ hybridization (CARD-FISH) detection of Dehalococcoides. AB - Members of the genus Dehalococcoides are well-known for their capacity to reductively dechlorinate chlorinated organic pollutants. The availability of quantitative and sensitive detection methods is of major interest for research on the ecology of those environmentally important micro-organisms. In this paper we describe the development of a Catalyzed Reporter Deposition-Fluorescent In Situ Hybridization (CARD-FISH) for detection of Dehalococcoides cells in enrichment cultures using two oligonucleotide sequences which target two different lineages of Dehalococcoides as probes. Both sequences were previously applied in conventional FISH as probes. Conjugation of the probe to horseradish peroxidase (HRP) did not change the specificity of the probes and bright fluorescent signals were obtained. Despite the use of higher concentrations of probe and the application of longer exposure times in the conventional FISH procedure, CARD FISH resulted in more intense signals. The CARD-FISH method was applied to a vinyl chloride (VC)-reductively-dechlorinating enrichment culture. Only the probe targeting the CBDB1 lineage of Dehalococcoides reacted with the sample which was in agreement with previous nucleic acid based analysis. The culture consisted of 51%+/-8% of Dehalococcoides cells. Furthermore, the CARD-FISH probes for detecting Dehalococcoides were combined with FISH probes for simultaneous detection of either Bacteria or Archaea which should allow rapid insight into the relative dynamics of the different members of dechlorinating communities as a response to environmental changes. Overall, CARD-FISH proved to be a rapid, reliable and convenient method to detect and quantify Dehalococcoides cells. PMID- 18410974 TI - Rapid isolation of host-independent Bdellovibrio bacteriovorus. AB - A new method of isolating host-independent Bdellovibrio bacteriovorus has been developed. Filtered suspensions of host-dependent cells are dropped in small volumes onto 0.2 microm membranes laid on rich media agar. Significant growth is observed within 1-2 days; these cells were confirmed to be B. bacteriovorus using microscopic observations and PCR. PMID- 18410975 TI - Pulsed dose rate brachytherapy as the sole adjuvant radiotherapy after breast conserving surgery of T1-T2 breast cancer: first long time results from a clinical study. AB - BACKGROUND AND PURPOSE: To evaluate the long time outcome with regard to local tumour control, cosmetic outcome and side effects of a short (5 days) accelerated interstitial brachytherapy treatment delivered to the surroundings of the operated sector. PATIENTS AND METHODS: Between 1993 and 2003 we treated 50 women with early T1 and T2 breast cancer. Radical sector resection was performed and followed later with an interstitial pulsed dose rate (PDR) brachytherapy of 50Gy in 5 days. The treatment was centred on the tumour with a margin of 30mm. One patient was treated bilaterally. The patients were followed for a median of 86 (32-126) months. RESULTS: Ipsilateral breast cancer recurrence was seen in 3 patients (6%). Two of them occurred outside the treated volume. The 5- and 7-year rates of actuarial local control were 96% and 96%, respectively, overall survival 88% and 85%, disease free survival 88% and 88%, respectively. A dosimetrical analysis showed that the partial breast irradiation covered a median of 31% of the total breast volume. Fat necrosis was seen in 12% and local (moderate-strong) fibrosis in 26% of the patients. Independent cosmetic scoring showed good or excellent result in 56% of the patients. CONCLUSIONS: Local outcome is favourable and very similar to other published studies of accelerated partial breast irradiation. Our long time cosmetic results are lower than other published results. PMID- 18410976 TI - Predictors of acute grade 4 swallowing toxicity in patients with stages III and IV squamous carcinoma of the head and neck treated with radiotherapy alone. AB - PURPOSE: The purpose of the study was to investigate the predictive factors for acute grade 4 swallowing toxicity in an attempt to identify which patients may benefit from early intervention with enteral feeding during curative radiation treatment for localised Stages 3-4 squamous cell carcinoma of the head and neck. It was hypothesised that craniocaudal length of the treatment field to the upper neck and pharynx would correlate with grade 4 swallowing toxicity due to the increased volume of pharynx irradiated. PATIENTS AND METHODS: Toxicity data were collected prospectively as part of a phase III randomised trial (TROG 91:01) that assigned patients to either conventional (CRT) or accelerated radiotherapy (ART). Patients were randomly assigned to either CRT, using a single 2 Gy per day to a dose of 70 Gy in 35 fractions in 49 days or to ART, using 1.8 Gy twice a day to a dose of 59.4 Gy in 33 fractions in 24 days. Treatment allocation was stratified for site and stage. Accrual commenced in 1991 and the trial was closed in 1998 when the target of 350 patients was reached. Potential factors were analysed that predicted for Grade IV swallowing toxicity. RESULTS: The treatment field lengths >82mm for the second phase increased the probability of requiring intervention with percutaneous endoscopic gastrostomy (PEG) or Nasogastric tube (NGT). The probability of grade 4 swallowing was 36% if the phase 2 treatment length was >82mm vs 16% for less < or = 82mm(p=0.0001). A predictive enteral grading score (PEG score) was derived using the Cox regression coefficients: Field length of the boost volume >82mm scored 3 points, Stage grouping greater than 1 scored 1 point, altered fractionation scored 2 points, ECOG greater than 1 scored 1 point. The PEG score was 45% if the score was 6 and 19% if the score was <6 (p=0.0). CONCLUSIONS: More attention needs to be focused on developing robust dose and volume constraints for the pharyngeal mucosa and the musculature in order to reduce the need for enteral feeding. Patients with PEG score of 6 or greater are at high risk of requiring enteral feeding during radiation treatment and should be considered for prophylactic PEG or NG feeding. PMID- 18410977 TI - Screening for distant metastases in head and neck cancer patients by chest CT or whole body FDG-PET: a prospective multicenter trial. AB - BACKGROUND AND PURPOSE: The aim of the study was to define the added value of whole body FDG-PET in screening for distant metastases in patients with head and neck squamous cell carcinoma and risk factors. MATERIALS AND METHODS: In a multi center prospective study between 1998 and 2003, 145 consecutive HNSCC patients with risk factors for distant metastases underwent chest CT and whole body FDG PET for screening of distant metastases. The data of 92 evaluable patients who developed distant metastases or who had a follow-up of at least 12 months were analyzed. Besides their performance in clinical practice, the operational characteristics of PET and CT using ROC analyses were investigated. RESULTS: Pretreatment screening identified distant metastases in 19 patients (21%). FDG PET had a higher sensitivity (53% vs. 37%) and positive predictive value (80% vs. 75%) than CT. The combination of CT and FDG-PET had the highest sensitivity (63%). The ROC analyses of the five point ordinal scales revealed that the "area under the curve" (AUC) of FDG-PET was significantly higher as compared to CT. CONCLUSION: In HNSCC patients with risk factors, pretreatment screening for distant metastases by chest CT is improved by FDG-PET. PMID- 18410978 TI - Proceedings of the International Symposium on Vinegars and Acetic Acid Bacteria 2005. May 8-12, 2005. Reggio Emilia, Italy. PMID- 18410979 TI - The H syndrome: a genodermatosis characterized by indurated, hyperpigmented, and hypertrichotic skin with systemic manifestations. AB - BACKGROUND: The association of cutaneous hyperpigmented, hypertrichotic, and indurated patches associated with hearing loss, short stature, cardiac anomalies, hepatosplenomegaly, scrotal masses, and hypogonadism has not, to our knowledge, been previously recognized as a disease entity. OBJECTIVE: We describe 10 patients with the above-mentioned findings. METHODS: Patients were clinically examined and extensive laboratory evaluation was performed. RESULTS: We describe 10 patients from 6 Arab consanguineous families with hyperpigmented, hypertrichotic, and indurated cutaneous patches involving the middle and lower parts of their bodies. In addition, patients displayed short stature, sensorineural hearing loss, cardiac anomalies, hepatosplenomegaly, and scrotal masses. Laboratory evaluation revealed growth hormone deficiency and hypergonadotropic hypogonadism with azoospermia. Cutaneous histopathologic examination showed hyperpigmentation of the basal layer with seborrheic-keratosis like acanthosis, histiocytic infiltration, and a perivascular mononuclear infiltrate with plasma cells and mast cells throughout the dermis and subcutaneous fat. Comparison with several patients, recently reported in the medical literature, with similar cutaneous findings is made. LIMITATIONS: Laboratory evaluation in some patients was incomplete because of lack of cooperation. CONCLUSIONS: We suggest that our patients represent a novel multisystemic autosomal recessive inherited disorder. We call this constellation of symptoms the "H syndrome." PMID- 18410980 TI - Safety of dermatologic drugs used in pregnant patients with psoriasis and other inflammatory skin diseases. AB - In patients with psoriasis, there is an increased availability of drugs for treatment. However, there are important questions about drug safety for mothers with psoriasis and their fetuses. Currently, there are limited safety data for many of the medications used. In this article, we review current pregnancy risk information for medications commonly used in the treatment of psoriasis. In addition, a list of teratology information resources is included to help practicing clinicians find up-to-date information regarding the safety of the medications they prescribe. PMID- 18410981 TI - Drug-induced morphea: report of a case induced by balicatib and review of the literature. AB - Drug-induced scleroderma has been rarely reported, mostly with the features of diffuse scleroderma or acrosclerosis, and exceptionally with the characteristics of morphea. We report the case of an adult white woman, enrolled in a double blind, placebo-controlled, multicentric trial evaluating the efficacy and safety of the cathepsin K inhibitor balicatib for osteoporosis. Typical morphea lesions developed on the patient's trunk 9 months after the beginning of therapy. Lesions completely resolved after drug withdrawal and a single brief course of systemic steroids. No recurrence occurred in a 2-year follow-up. Fifteen cases of drug induced morphea could be retrieved from the literature. Drug withdrawal determined complete remission in only a few patients. Different drug classes have been implicated. Some of these, including balicatib, alter directly connective tissue metabolism. PMID- 18410982 TI - Synthesis and use of 4-peptidylhydrazido-N-hexyl-1,8-naphthalimides as fluorogenic histochemical substrates for dipeptidyl peptidase IV and tripeptidyl peptidase I. AB - Gly-Pro-, Gly-Pro-Met- and Ala-Ala-Phe-N'-(2-hexyl-1,3-dioxo-2,3-dihydro-1H benzo[de]isoquinolin-6-yl)-hydrazides are synthesized by guanidinium/uronium type condensing reagent and used as fluorogenic substrates to localize dipeptidyl peptidase IV and tripeptidyl peptidase I activities in mammalian tissue sections. Enzyme hydrolysis releases 2-hexyl-6-hydrazino-1H-benzo[de]isoquinoline-1,3(2H) dione, which couples with piperonal to form insoluble fluorescent hydrazone, precipitating on the enzyme locations and marking them. The fluorescent technique reveals precisely the enzymes locations at the lack of background noise in a single incubation step. It avoids most of the drawbacks of the previously proposed fluorescent histochemical techniques and can be valuable for the in situ studies of these enzymes in norm and pathology. PMID- 18410983 TI - Effects of air pollution from road transport on growth and physiology of six transplanted bryophyte species. AB - Motor vehicles emit a cocktail of pollutants; however, little is known about the effects of these pollutants on bryophytes located in roadside habitats. Six bryophyte species were transplanted to either a woodland or a moorland site adjacent to a motorway, and were monitored over sevenmonths from autumn through to spring. All species showed an increase in one or more of the following near the motorway: growth, membrane leakage, chlorophyll concentration, and nitrogen concentration. The strongest effects were observed in the first 50-100 m from the motorway: this was consistent with the nitrogen dioxide pollution profile, which decreased to background levels at a distance of 100-125 m. It is hypothesised that motor vehicle pollution was responsible for the effects observed, and that nitrogen oxides had a key influence. The observed effects may lead to changes in vegetation composition with significant implications for nature conservation and management of roadside sites. PMID- 18410984 TI - Psychological approaches in the treatment of specific phobias: a meta-analysis. AB - Data from 33 randomized treatment studies were subjected to a meta-analysis to address questions surrounding the efficacy of psychological approaches in the treatment of specific phobia. As expected, exposure-based treatment produced large effects sizes relative to no treatment. They also outperformed placebo conditions and alternative active psychotherapeutic approaches. Treatments involving in vivo contact with the phobic target also outperformed alternative modes of exposure (e.g., imaginal exposure, virtual reality, etc.) at post treatment but not at follow-up. Placebo treatments were significantly more effective than no treatment suggesting that specific phobia sufferers are moderately responsive to placebo interventions. Multi-session treatments marginally outperformed single-session treatments on domain-specific questionnaire measures of phobic dysfunction, and moderator analyses revealed that more sessions predicted more favorable outcomes. Contrary to expectation, effect sizes for the major comparisons of interest were not moderated by type of specific phobia. These findings provide the first quantitative summary evidence supporting the superiority of exposure-based treatments over alternative treatment approaches for those presenting with specific phobia. Recommendations for future research are also discussed. PMID- 18410985 TI - Categorical and dimensional approaches to the measurement of disruptive behavior in the preschool years: a meta-analysis. AB - Preschool disruptive behavior problems were investigated in a meta-analysis of 26 studies using categorical and/or dimensional approaches to assessment. The review sought to distinguish early disruptiveness from normative preschool conduct by showing that, irrespective of assessment methodology: (a) disruptiveness can be adequately measured in the preschool years; (b) early disruptiveness is stable over time; and (c) disruptive children referred for clinical services in the preschool years can be distinguished from non-referred peers. Results indicated that: categorical and dimensional approaches to measurement of early disruptiveness provide comparable data (effect size d=2.29); both approaches yield comparable estimates of the stability of preschool disruptive behavior over time (categorical approach: d=1.15; dimensional approach: d=0.84); and both approaches discriminate between referred and non-referred preschoolers (d=1.05 and d=.95). Limitations of the existing literature and of this analysis are discussed, as are suggestions for future research. PMID- 18410986 TI - Stigma, social inequality, and HIV risk disclosure among Dominican male sex workers. AB - Some quantitative behavioral studies in the USA have concluded that bisexually behaving Latino men are less likely than White men to disclose to their female partners that they have engaged in same-sex risk behavior and/or are HIV positive, presumably exposing female partners to elevated risk for HIV infection. Nevertheless, very little theoretical or empirical research has been conducted to understand the social factors that promote or inhibit sexual risk disclosure among Latino men who have sex with men (MSM), and much of the existing literature has neglected to contextualize disclosure patterns within broader experiences of stigma and social inequality. This paper examines decisions about disclosure of sex work, same-sex behavior, and sexual risk for HIV among male sex workers in two cities in the Dominican Republic. Data derive from long-term ethnography and qualitative in-depth interviews with 72 male sex workers were used to analyze the relationships among experiences of stigma, social inequality, and patterns of sexual risk disclosure. Thematic analysis of interviews and ethnographic evidence revealed a wide range of stigma management techniques utilized by sex workers to minimize the effects of marginality due to their engagement in homosexuality and sex work. These techniques imposed severe constraints on men's sexual risk disclosure, and potentially elevated their own and their female partners' vulnerability to HIV infection. Based on the study's findings, we conclude that future studies of sexual risk disclosure among ethnic minority MSM should avoid analyzing disclosure as a decontextualized variable, and should seek to examine sexual risk communication as a dynamic social process constrained by hierarchical systems of power and inequality. PMID- 18410987 TI - Transcriptional regulation of bone marrow thrombopoietin by platelet proteins. AB - Platelet production is regulated primarily by the cytokine thrombopoietin (TPO). Although TPO is expressed in several different tissues, only in the bone marrow has the level of expression been reported to increase in response to reduced numbers of platelets. In these studies, we demonstrate that platelet granule proteins are able to transcriptionally repress TPO mRNA expression in a marrow stromal cell line as well as in primary bone marrow stromal cell cultures. Like TPO mRNA, secretion of TPO protein was also suppressed by serum treatment. Reporter gene constructs indicate that DNA elements located in an approximately 1.9-kb region between 250-bp upstream of the transcriptional initiation site and the middle of the second intron are able to mediate the transcriptional repression. PMID- 18410988 TI - Stem cell plasticity: clinical implications. PMID- 18410989 TI - Combined Th2 cytokine deficiency in donor T cells aggravates experimental acute graft-vs-host disease. AB - The role of T helper (Th) 1 and Th2 polarization in acute graft-vs-host-disease (GVHD) is unclear. We investigated the role of Th2 cytokine secretion by utilizing donor T cells that cannot make interleukin (IL)-4, IL-5, IL-9, and IL 13 from quadruple cytokine-deficient (Quad-KO) animals in a well-characterized BALB/c-->C57BL/6 model of allogeneic bone marrow transplantation. B6 recipients of BALB/c Quad-KO T cells demonstrated greater clinical severity, target organ damage, and mortality from GVHD than recipients of BALB/c wild-type (WT) T cells. When compared with donor T cells that are deficient in signal transducers and activators of transcription 6 signaling or the signature Th2 cytokine, IL-4, Quad KO T cells demonstrated greater GVHD mortality. Mechanistic studies demonstrated that Quad-KO T cells demonstrated enhanced T-cell proliferation than WT T cells when stimulated with either allogeneic antigen-presenting cells or with nonspecific stimuli, such as anti-CD3 monoclonal antibody. Quad-KO T cells also secreted greater amounts of Th1 cytokines and IL-17 compared to WT T cells. Deficiency of Th2 cytokines, however, did not alter the allospecific cytotoxic responses, the numbers of immunoregulatory CD4(+)CD25(+) Foxp3(+) T cells or their suppressive functions. Our data thus unequivocally demonstrate that deficiency of the four classical Th2 cytokine enhances T-cell proliferative responses and aggravates GVHD. PMID- 18410990 TI - Suspended sediment, nitrogen and phosphorus concentrations and exports during storm-events to the Tuross estuary, Australia. AB - This paper presents a process for estimating pollutant loads from water quality data, to improve catchment-scale modelling in the region for resource management purposes. It describes a program to estimate suspended sediment, total and dissolved nitrogen and phosphorus loads to the Tuross estuary from the Tuross River catchment (1810 km(2)) of coastal southeast Australia. Event-based water quality sampling results obtained during storm events in 2005 are presented. Event 1, during July 2005 was the largest storm event in terms of peak flow for 3.5 years. Other events monitored were also in July, November and December 2005. The early July 2005 event had a flow-weighted mean suspended sediment (SS) concentration during the first 4 days of 63 mg L(-1). Of the events monitored, this was unusual as it was preceded by drought and had the largest SS concentrations (peaking at 180 mg L(-1)) during the rising-stage. In contrast, the November event had a much lower flow-weighted SS mean (28 mg L(-1)), even though peak flow magnitudes were similar. The July and November 2005 events had peak flows of 12,360 and 11,330 ML day(-1). Low-cost rising-stage siphon samplers were used to collect samples during the rapidly rising phase of these events. The use of such samplers and consideration of time-lead/lag flow adjustments, quantified using cross-correlation analysis to account for hysteresis effects, were incorporated into the load estimation techniques. The technique is a potentially useful approach for understanding relationships between water quality concentrations and flow for modelling catchment source strengths and transport processes. PMID- 18410991 TI - Chronic pain may change the structure of the brain. AB - Recently, local morphologic alterations of the brain in areas ascribable to the transmission of pain were detected in patients suffering from phantom pain, chronic back pain, irritable bowl syndrome, fibromyalgia and two types of frequent headaches. These alterations were different for each pain syndrome, but overlapped in the cingulate cortex, the orbitofrontal cortex, the insula and dorsal pons. These regions function as multi-integrative structures during the experience and the anticipation of pain. As it seems that chronic pain patients have a common "brain signature" in areas known to be involved in pain regulation, the question arises whether these changes are the cause or the consequence of chronic pain. The author suggests that the gray matter change observed in chronic pain patients are the consequence of frequent nociceptive input and should thus be reversible when pain is adequately treated. PMID- 18410992 TI - Pain and sleep: establishing bi-directional association in a population-based sample. PMID- 18410993 TI - Potential therapeutic usefulness of hydrogen peroxide in conditions of brain ischemia. PMID- 18410994 TI - The role of the basal ganglia in data processing. AB - Complex cerebral activities are likely to be composed of massively repeated simple data processing tasks since the cortical data processing unit, the cortical mini-column, is found throughout the cortex with only minor variations. It has been proposed that one task performed by the cortical mini-column may be to match afferent sensory data to learnt datasets in a process known as automatic association. We hypothesize that basal ganglia circuits, through the relative signal of the nigro-striatal and striato-pallidal pathways, determine the matching threshold for dataset matching within cortical mini-columns. Basal ganglia circuits are in a unique position to use parallel information to modulate the parameters of auto-association to increase the speed of data processing tasks. This hypothesis can explain motor symptoms in Parkinson's disease and also predicts that over and underactivity of basal ganglia circuits (the 'on' and 'off' states) will lead to characteristic errors in sensory data interpretation in all modalities - false negative data recognition when 'off' and false positive data recognition when 'on'. As a preliminary exploration of this hypothesis 16 patients with advanced Parkinson's disease were tested in voice and face recognition when 'off' and 'on'. Each patient exhibited errors in the recognition task according to basal ganglia activity as predicted by our hypothesis. Further experiments to test the hypothesis are proposed. PMID- 18410995 TI - Molecular evidence demonstrating local treatment failure is the source of distant metastases in some patients treated for breast cancer. AB - PURPOSE: To examine the clonality relationships among initial invasive breast carcinoma (IBC), ipsilateral breast failure (IBF), and distant metastasis (DM) to determine the effect of local tumor recurrence on the development of DMs. METHODS AND MATERIALS: A total of 18 patients treated with breast-conserving therapy who developed an IBF followed by DMs were studied using a 20 informative-marker, polymerase chain reaction-based allelic imbalance clonality assay. RESULTS: Four relationships were identified. First, in 7 cases, the IBF and DMs were clonally related to the initial IBC as one progressively genetic unstable process. Second, in 3 cases, the IBF and DMs were each clonally related to the IBC but clonally distinct from each other. Third, in 3 cases, the IBC and the IBF were clonally related and the DMs were clonally related to the IBFs, with a weak relationship to the initial IBC. Finally, in 5 cases, the IBF was clonally distinct from the initial IBC (new second primary) and the DMs were clonally related to the IBF and clonally distinct from the initial IBC. CONCLUSION: These findings provide molecular evidence demonstrating that some DMs can directly develop from IBFs and support the importance of local tumor control in the overall treatment of breast cancer patients. PMID- 18410996 TI - Radiosensitization of human vascular endothelial cells through Hsp90 inhibition with 17-N-allilamino-17-demethoxygeldanamycin. AB - PURPOSE: In addition to invasive tumor cells, endothelial cells (ECs) of the tumor vasculature are an important target for anticancer radiotherapy. The purpose of the present work is to investigate how 17-N-allilamino-17 demethoxygeldanamycin (17AAG), known as an anticancer drug inhibiting heat shock protein 90 (Hsp90), modifies radiation responses of human vascular ECs. METHODS AND MATERIALS: The ECs cultured from human umbilical veins were exposed to gamma irradiation, whereas some EC samples were pretreated with growth factors and/or 17AAG. Postirradiation cell death/survival and morphogenesis were assessed by means of terminal deoxynucleotidyl transferase biotin-deoxyuridine triphosphate nick end labeling or annexin V staining and clonogenic and tube-formation assays. The 17AAG-affected expression and phosphorylation of radioresistance-related proteins were probed by means of immunoblotting. Dominant negative or constitutively activated Akt was transiently expressed in ECs to manipulate Akt activity. RESULTS: It was found that nanomolar concentrations of 17AAG sensitize ECs to relatively low doses (2-6 Gy) of gamma-irradiation and abolish the radioprotective effects of vascular endothelial growth factor and basic fibroblast growth factor. The drug-induced radiosensitization of ECs seems to be caused by prevention of Hsp90-dependent phosphorylation (activation) of Akt that results in blocking the radioprotective phosphatidylinositol 3-kinase/Akt pathway. CONCLUSIONS: Clinically achievable concentrations of 17AAG can decrease the radioresistance intrinsic to vascular ECs and minimize the radioprotection conferred upon them by tumor-derived growth factors. These findings characterize 17AAG as a promising radiosensitizer for the tumor vasculature. PMID- 18410998 TI - Is a loose-seed nomogram still valid for prostate brachytherapy in a stranded seed era? AB - PURPOSE: To characterize the amount of activity required to treat the prostate with stranded (125)I radioactive seeds and compare our stranded data with the amount of activity recommended when individual seeds are implanted using a Mick applicator. METHODS AND MATERIALS: Data from two groups of patients at The University of Texas M. D. Anderson Cancer Center who were treated with prostate brachytherapy as monotherapy were analyzed. The first group included 100 patients implanted with individual seeds in 2000 and 2001. The second group comprised 81 patients for whom stranded seeds were implanted in 2006 and 2007. Seeds in both groups were (125)I seeds with an air kerma strength of 0.497 U per seed (0.391 mCi per seed). The prescribed dose to planning target volume was 145 Gy. RESULTS: The total implanted activity and the number of seeds used were significantly lower in the second group (p < 0.0001) than in the first group. The reduction in activity in the stranded-seed group was approximately 23% for a 20-cm(3) prostate and approximately 15% for a 60-cm(3) prostate. With equivalent activity between the two groups, the stranded-seed treatment covered a larger treatment volume with the prescribed dose. CONCLUSIONS: The amount of activity required to effectively treat a prostate of a given volume was lower with stranded seeds than with loose seeds. Our experience suggests that prostate brachytherapy that uses stranded seeds leads to a more efficient implant with fewer seeds and lower overall activity, resulting in improved homogeneity. PMID- 18410997 TI - Parapharyngeal extension of nasopharyngeal carcinoma: still a significant factor in era of modern radiotherapy? AB - PURPOSE: To retrospectively analyze the prognostic value of parapharyngeal space (PPS) extension after conformal radiotherapy for nasopharyngeal carcinoma. PATIENTS AND METHODS: Between 1998 and 2005, 700 patients were treated with conformal radiotherapy at 2 Gy/fraction daily to a total of 70 Gy. All patients underwent staging with magnetic resonance imaging. The incidence of PPS was determined, and the extent of involvement was further subclassified regarding the presence or absence of carotid space (CS) involvement. The prognostic parameters, including age, gender, stage, chemotherapy, additional boosting, and extent of PPS involvement, were analyzed by univariate and multivariate analyses. RESULTS: The median duration of follow-up was 51 months, and the 5-year overall survival rate for the whole group was 73%. The overall incidence of PPS extension was high (74%), and 29% had additional extension to the CS. Multivariate analysis showed age, gender, chemotherapy, T stage, and N stage to be significant prognostic factors, but not PPS involvement with or without CS extension. In the subgroup of patients with Stage T2 disease (n = 242), the presence of PPS involvement alone or PPS plus CS extension had no statistically significant effect in terms of local control (p = 0.68), distant metastases (p = 0.34), or overall survival (p = 0.24) compared with those without PPS involvement (Stage T2a). CONCLUSIONS: With better tumor delineation by magnetic resonance imaging and improved coverage using modern radiotherapy techniques, PPS extension per se no longer predicts for disease outcome. Hence, subcategorizing Stage T2 disease is no longer important in future International Union Against Cancer/American Joint Committee on Cancer classifications. PMID- 18410999 TI - Hdm2 and nitric oxide radicals contribute to the p53-dependent radioadaptive response. AB - PURPOSE: The aim of this work was to characterize the radioadaptive response at the molecular level. METHODS AND MATERIALS: We used wild-type (wt) p53 and mutated (m) p53-containing cells derived from the human lung cancer H1299 cell line, which is p53-null. Cellular radiation sensitivities were determined with a colony-forming assay. The accumulations of p53, the human homolog of endogenous murine double minute 2 (Hdm2), and inducible nitric oxide synthase were analyzed with Western blotting. Quantification of chromosomal aberrations was estimated by scoring dicentrics per cell. RESULTS: In wtp53 cells, it was demonstrated that the lack of p53 accumulation was coupled with the activation of Hdm2 after low dose irradiation (0.02 Gy). Although NO radicals were only minimally induced in wtp53 cells irradiated with a challenging irradiation (6 Gy) alone, NO radicals were seen to increase about two- to fourfold after challenging irradiation subsequent to a priming irradiation (0.02 Gy). Under similar irradiation conditions with a priming and challenging irradiation in wtp53 cells, induction of radioresistance and a depression of chromosomal aberrations were observed only in the absence of 5, 5'-(2, 5-Furanidiyl)bis-2-thiophenemethanol (RITA) or Nutlin 3 (p53-Hdm2 interaction inhibitors), aminoguanidine (an inducible nitric oxide synthase inhibitor), and c-PTIO (an NO radical scavenger). On the other hand, in p53 dysfunctional cells, a radioadaptive response was not observed in the presence or absence of those inhibitors. Moreover radioresistance developed when wtp53 cells were treated with isosorbide dinitrate (an NO-generating agent) alone. CONCLUSIONS: These findings suggest that NO radicals are initiators of the radioadaptive response, acting through the activation of Hdm2 and the depression of p53 accumulations. PMID- 18411000 TI - Reporting late rectal toxicity in prostate cancer patients treated with curative radiation treatment. AB - PURPOSE: Long-term rectal toxicity is a concern for patients with prostate cancer treated with curative radiation. However, comparing results of late toxicity may not be straightforward. This article reviews the complexity of reporting long term side effects by using data for patients treated in our institution with hypofractionated irradiation. METHODS AND MATERIALS: Seventy-two patients with localized prostate cancer treated with hypofractionated radiotherapy alone to a dose of 66 Gy in 22 fractions were prospectively assessed for late rectal toxicity according to the Common Toxicity Criteria, Version 3, scoring system. Ninety percent of patients had more than 24 months of follow-up. Results are compared with data published in the literature. RESULTS: We found an actuarial incidence of Grade 2 or higher late rectal toxicity of 27% at 30 months and a crude incidence of Grade 2 or higher late rectal toxicity of 18%. This was mostly severe toxicity documented during follow-up. The incidence of Grade 3 rectal toxicity at the last visit was 3% compared with 13% documented at any time during follow-up. CONCLUSION: Comparison of late toxicity after radiotherapy in patients with prostate cancer must be undertaken with caution because many factors need to be taken into consideration. Because accurate assessment of late toxicity in the evaluation of long-term outcome after radiotherapy in patients with localized prostate cancer is essential, there is a need to develop by consensus guidelines for assessing and reporting late toxicity in this group of patients. PMID- 18411001 TI - Prospective evaluation of pretreatment executive cognitive impairment and depression in patients referred for radiotherapy. AB - PURPOSE: Cancer patients are at risk of cognitive impairment and depression. We sought to ascertain the prevalence of executive, visuospatial, memory, and general cognitive performance deficits before radiotherapy in a radiation oncology clinic referral population and correlate the neurocognitive measures with the depression symptom burden. METHODS AND MATERIALS: A total of 122 sequential patients referred for radiotherapy evaluation were administered a test battery composed of the Executive Interview (EXIT25), Executive Clock Drawing Task (CLOX1 and CLOX2), Mini Mental State Examination (MMSE), Memory Impairment Screen (MIS), and Geriatric Depression Scale (GDS). The mean age +/- standard deviation was 58 +/- 17 years. Of 122 patients, 24 (20%) had been referred for breast cancer, 21 (17%) for gastrointestinal cancer, 17 (14%) for genitourinary disease, and 8 (7%) for brain lesions; the rest were a variety of tumor sites. The cognitive performance among the tumor cohorts was compared using Bonferroni corrected analysis of variance and Tukey-Kramer tests. Pearson correlation coefficients were determined between each cognitive instrument and the GDS. RESULTS: Of the 122 patients, 52 (43%) exhibited a detectable executive cognition decrement on one or more test measures. Five percent had poor memory performance (MIS), 18% had poor visuospatial performance (CLOX2), and 13% had poor global cognition (MMSE). Patients with brain tumors performed substantially worse on the EXIT25. No between-group differences were found for CLOX1, CLOX2, MIS, or GDS performance. The EXIT25 scores correlated significantly with the GDS scores (r = 0.26, p = 0.005). CONCLUSIONS: The results of this study have shown that patients referred for radiotherapy exhibit cognitive impairment profiles comparable to those observed in acutely ill medical inpatients. Executive control impairment appears more prevalent than global cognitive deficits, visuospatial impairment, or depression. PMID- 18411002 TI - Small molecular inhibitor of transforming growth factor-beta protects against development of radiation-induced lung injury. AB - PURPOSE: To determine whether an anti-transforming growth factor-beta (TGF-beta) type 1 receptor inhibitor (SM16) can prevent radiation-induced lung injury. METHODS AND MATERIALS: One fraction of 28 Gy or sham radiotherapy (RT) was administered to the right hemithorax of Sprague-Dawley rats. SM16 was administered in the rat chow (0.07 g/kg or 0.15 g/kg) beginning 7 days before RT. The rats were divided into eight groups: group 1, control chow; group 2, SM16, 0.07 g/kg; group 3, SM16, 0.15 g/kg; group 4, RT plus control chow; group 5, RT plus SM16, 0.07 g/kg; group 6, RT plus SM16, 0.15 g/kg; group 7, RT plus 3 weeks of SM16 0.07 g/kg followed by control chow; and group 8, RT plus 3 weeks of SM16 0.15 g/kg followed by control chow. The breathing frequencies, presence of inflammation/fibrosis, activation of macrophages, and expression/activation of TGF-beta were assessed. RESULTS: The breathing frequencies in the RT plus SM16 0.15 g/kg were significantly lower than the RT plus control chow from Weeks 10-22 (p <0.05). The breathing frequencies in the RT plus SM16 0.07 g/kg group were significantly lower only at Weeks 10, 14, and 20. At 26 weeks after RT, the RT plus SM16 0.15 g/kg group experienced a significant decrease in lung fibrosis (p = 0.016), inflammatory response (p = 0.006), and TGF-beta1 activity (p = 0.011). No significant reduction was found in these measures of lung injury in the group that received SM16 0.7 g/kg nor for the short-course (3 weeks) SM16 at either dose level. CONCLUSION: SM16 at a dose of 0.15 g/kg reduced functional lung damage, morphologic changes, inflammatory response, and activation of TGF-beta at 26 weeks after RT. The data suggest a dose response and also suggest the superiority of long-term vs. short-term dosing. PMID- 18411003 TI - Arterial chemoradiotherapy for locally advanced tongue cancer: analysis of retrospective study of therapeutic results in 88 patients. AB - PURPOSE: To retrospectively investigate the therapeutic results of arterial injection therapy by way of the superficial temporal artery for 88 cases of Stage III and IV (M0) tongue cancer and to clarify the factors that affected the therapeutic results. METHODS AND MATERIALS: We administered intra-arterial chemoradiotherapy by continuous infusion of carboplatin in 39 patients between January 1993 and July 2002. Systemic concurrent chemotherapy was given to 19 of these patients. We administered intra-arterial chemoradiotherapy with cisplatin with sodium thiosulfate to 49 patients between October 2002 and December 2006. Concurrent systemic chemotherapy was given to 38 of these patients. RESULTS: The 3-year local control rate was 72% (T2-T3, 80%; and T4, 48%), and the 3-year survival rate was 57% (Stage III, 67%; Stage IV, 43%). On univariate analysis, age, T stage, N stage, overall stage, systemic chemotherapy, difference in intra arterial chemotherapy, and performance status had a significant effect on survival. On multivariate analysis, N stage, systemic chemotherapy, difference in intra-arterial chemotherapy, and artery selected had a significant effect on survival. CONCLUSIONS: The therapeutic results of intra-arterial chemoradiotherapy using the superficial temporal artery were not inferior to the results of surgery. In particular, the results of arterial injection therapy using cisplatin with sodium thiosulfate were excellent, and we believe it will be a new therapy for advanced tongue cancer. PMID- 18411004 TI - Low-dose radiation potentiates the therapeutic efficacy of folate receptor targeted hapten therapy. AB - PURPOSE: Human cancers frequently overexpress a high-affinity cell-surface receptor for the vitamin folic acid. Highly immunogenic haptens can be targeted to folate receptor-expressing cell surfaces by administration of folate-hapten conjugates, rendering the decorated tumor cell surfaces more recognizable by the immune system. Treatment of antihapten-immunized mice with folate-hapten constructs results in elimination of moderately sized tumors by the immune system. However, when subcutaneous tumors exceed 300 mm(3) before initiation of therapy, antitumor activity is significantly decreased. In an effort to enhance the efficacy of folate-targeted hapten immunotherapy (FTHI) against large tumors, we explored the combination of targeted hapten immunotherapy with low-dose radiotherapy. METHODS AND MATERIALS: Mice bearing 300-mm(3) subcutaneous tumors were treated concurrently with FTHI (500 nmol/kg of folate conjugated to fluorescein isothiocyanate, 20,000 U/dose of interleukin 2, and 25,000 U/dose of interferon alpha) and low-dose radiotherapy (3 Gy/dose focused directly on the desired tumor mass). The efficacy of therapy was evaluated by measuring tumor volume. RESULTS: Tumor growth analyses show that radiotherapy synergizes with FTHI in antihapten-immunized mice, thereby allowing for cures of animals bearing tumors greater than 300 mm(3). More importantly, nonirradiated distal tumor masses in animals containing locally irradiated tumors also showed improved response to hapten immunotherapy, suggesting that not all tumor lesions must be identified and irradiated to benefit from the combination therapy. CONCLUSIONS: These results suggest that simultaneous treatment with FTHI and radiation therapy can enhance systemic antitumor activity in tumor-bearing mice. PMID- 18411005 TI - When is CT-based postoperative seroma most useful to plan partial breast radiotherapy? Evaluation of clinical factors affecting seroma volume and clarity. AB - PURPOSE: To evaluate the effect of the time from surgery and other clinical factors on seroma volume and clarity and establish the optimal time to use the computed tomography (CT)-based seroma to plan partial breast irradiation (PBI). METHODS AND MATERIALS: A total of 205 women with early-stage breast cancer underwent planning CT after breast-conserving surgery. One radiation oncologist contoured the seroma volume and scored the seroma clarity, using a standardized Seroma Clarity Score scale, from 0 (not detectable) to 5 (clearest). Univariate and multivariate analyses were performed to evaluate the associations between the seroma characteristics and the interval from surgery and other clinical factors. RESULTS: The mean interval from surgery to CT was 84 days (standard deviation 59). During postoperative Weeks 3-8, the mean seroma volume decreased from 47 to 30 cm(3), stabilized during Weeks 9-14 (mean 21) and was involuted beyond 14 weeks (mean 9 cm(3)). The mean seroma clarity score was 3.4 at Weeks 3-8, 2.5 at Weeks 9-14, and 1.6 after 14 weeks. The seroma clarity was greater in patients aged >or=70 years. The seroma volume and clarity correlated significantly with the volume of excised breast tissue but not with the maximal tumor diameter, surgical re-excision, or chemotherapy use. CONCLUSION: The optimal time to obtain the planning CT scan for PBI is within 8 weeks after surgery. During Weeks 9-14, the seroma might remain adequately defined in some patients; however, after 14 weeks, alternate strategies are needed to identify the PBI target. The lack of correlation between the seroma volume and tumor size suggests that the CT-based seroma should not be the sole guide for PBI target volume definition. PMID- 18411006 TI - Ethanol inhibition of angiotensin II-stimulated Tyr705 and Ser727 STAT3 phosphorylation in cultured rat hepatocytes: relevance to activation of p42/44 mitogen-activated protein kinase. AB - Angiotensin (Ang) II-stimulated phosphorylation of signal transducer and activator transcription (STAT) 3 in rat hepatocytes and the effects of ethanol on this activation were investigated. Angiotensin II (100 nM) stimulated Tyr705 and Ser727 phosphorylation of STAT3 and formation of sis-inducing factor complexes. In the presence of U-0126 (10microM), a p42/44 mitogen-activated protein kinase (MAPK) kinase inhibitor, Ang II further increased Tyr705 phosphorylation of STAT3 but completely abrogated Ser727 phosphorylation of STAT3. Inhibition of p42/44MAPK also increased STAT3 DNA-binding activity. Pretreatment with ethanol (100mM) for 24h resulted in decrease in Tyr705 phosphorylation of STAT3 by ethanol alone and inhibition of Tyr705 phosphorylation of STAT3 stimulated by Ang II. Although ethanol potentiates Ang II stimulated p42/44 MAPK activation in hepatocytes, ethanol inhibited Ser727 phosphorylation of STAT3 stimulated by Ang II. Angiotensin II-stimulated STAT3-binding activity was not significantly affected by ethanol treatment. These results suggest a negative regulation of Ang II-stimulated STAT3 tyrosine phosphorylation and STAT3-binding activity through p42/44 MAPK activation in hepatocytes. However, ethanol modulation of Ang II stimulated STAT3 phosphorylation occurs by MAPK independent mechanisms. Ethanol potentiation of MAPK signaling without suppression of STAT3 function may modulate the course of alcoholic liver injury. PMID- 18411008 TI - Effects of various osmolarity on human red blood cells in terms of potassium efflux and hemolysis induced by free radicals. AB - OBJECTIVES: To study in an in vitro model of erythrocytes exposed to free radicals the effects of hyperosmolarity and hypoosmolarity on the induced potassium efflux and hemolysis. DESIGN SETTING: Erythrocytes were separated from plasma and suspended in 3 different phosphate buffer solutions (HYPO, ISO, and HYPER) containing, respectively, 100, 150 and 200 mmol/l of Na. Free radicals were generated from 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH). Potassium efflux (flame photometry) and hemolysis (Drabkin method) were measured. Measurements were expressed in % (versus total) and area under % versus time curves were calculated (% min). An ANOVA was used for statistical analysis. RESULTS: In presence of AAPH, hemolysis was significantly greater in HYPO (732.45+/-40% min) and lower in HYPER (578.97+/-15% min) as compared to ISO (608.30+/-42% min). Potassium efflux was significantly increased in HYPER (7508.04+/-85% min) as compared with HYPO (5308.74+/-62% min) and ISO (6561.39+/ 62% min). DISCUSSION: Our data suggest that hyperosmolarity increases the resistance of the red blood cells when exposed to free radicals as shown by the decreased hemolysis. In such conditions, the potassium efflux analysis appears to be inappropriate to evaluate the free radical effects on erythrocytes. PMID- 18411009 TI - Reliability, validity, and responsiveness of the daily sleep interference scale among diabetic peripheral neuropathy and postherpetic neuralgia patients. AB - To evaluate the psychometric characteristics of the Daily Sleep Interference Scale (DSIS) in patients with painful diabetic peripheral neuropathy (DPN) or postherpetic neuralgia (PHN), a post hoc secondary analysis of data from eight randomized clinical trials (four DPN and four PHN) was performed. Data were pooled within patient populations when assessment weeks were the same. The DSIS was completed by 1,124 DPN and 1,034 PHN patients. Patient-reported outcomes, including a Daily Pain Diary, the Short-Form McGill Pain Questionnaire, SF-36 Health Survey, Profile of Mood States, MOS-Sleep Scale (MOS-SS), EQ-5D, and Patient Global Impression of Change, were used to validate the DSIS. Test-retest reliability was high for both samples (intraclass correlation coefficient>0.90). The DSIS showed good construct validity, with moderate to high correlations between the DSIS and weekly mean pain scores (r=0.48-0.80), MOS-SS sleep disturbance subscale (r=0.45-0.64), MPQ-SF Pain Experience (r=0.37-0.61), and VAS (r=0.42-0.72). The DSIS showed good discriminant validity in both groups; high and low MOS-SS sleep disturbance groups had significantly different DSIS scores (P<0.001). DPN patients who improved minimally on the Patient Global Impression of Change and in pain scores improved 1.5-2 DSIS points on average; for PHN, patient scores improved an average of 1-2 points. The DSIS demonstrated robust test-retest reliability, good construct and discriminant validity and responsiveness in painful DPN and PHN patients. A 1-2 point change on the DSIS might be interpreted as an important difference. PMID- 18411007 TI - Exposure-dependent effects of ethanol on the innate immune system. AB - Extensive evidence indicates that ethanol (alcohol) has immunomodulatory properties. Many of its effects on innate immune response are dose dependent, with acute or moderate use associated with attenuated inflammatory responses, and heavy ethanol consumption linked with augmentation of inflammation. Ethanol may modify innate immunity via functional alterations of the cells of the innate immune system. Mounting evidence indicates that ethanol can diversely affect antigen recognition and intracellular signaling events, which include activation of mitogen activated protein kinases, and NFkappaB, mediated by Toll-like receptors, leading to altered inflammatory responses. The mechanism(s) underlying these changes may involve dose-dependent effects of ethanol on the fluidity of cell membrane, resulting in interference with the timely assembly or disassembly of lipid rafts. Ethanol could also modify cell activation by specific interactions with cell membrane molecules. PMID- 18411010 TI - Efficacy and safety of transdermal buprenorphine: a randomized, placebo controlled trial in 289 patients with severe cancer pain. AB - Strong opioids are recommended for treating severe cancer pain in the advanced stages of the disease. Few data are available concerning the efficacy of buprenorphine in cancer pain. We compared transdermal buprenorphine 70 microg/h (BUP TDS) to placebo in an enriched design study. Opioid-tolerant patients with cancer pain requiring strong opioids in the dose range of 90-150 mg/d oral morphine equivalents entered a two-week run-in phase, during which they were converted to BUP TDS. Patients who could be stabilized on BUP TDS were randomized to BUP TDS or placebo patch for a two-week maintenance phase. Rescue medication (buprenorphine sublingual tablets 0.2mg) was allowed as required. Response was defined as a mean pain intensity of <5 (0-10 scale) and a mean daily buprenorphine sublingual tablet intake of < or =2 tablets during the maintenance phase. Of 289 patients who entered the run-in phase, 100 discontinued treatment due to lack of efficacy or adverse events; 189 patients continued treatment in the maintenance phase (94 BUP TDS, 95 placebo), of whom 31 discontinued treatment (7 BUP TDS, 24 placebo). A significant difference in the number of treatment responders was observed: 70 BUP TDS (74.5%, 65.7-83.3) vs. 47 placebo (50%, 39.9 60.1) (P=0.0003). This result was supported by a lower daily pain intensity, lower intake of buprenorphine sublingual tablets and fewer dropouts in the BUP TDS group. The incidence of adverse events was slightly higher for BUP TDS. In conclusion, BUP TDS 70 microg/h is an efficacious and safe treatment for patients with severe cancer pain. PMID- 18411011 TI - Fatigue and its risk factors in cancer patients who seek emergency care. AB - Cancer patients visiting the emergency center (EC) are seldom assessed or treated for severe fatigue, a common symptom in sick patients due to acute medical conditions arising from cancer and cancer treatment. We provide a profile of cancer-related fatigue within the EC setting. Using a single-item screening tool derived from the Brief Fatigue Inventory, 928 patients (636 with solid tumors, 292 with hematological malignancies) triaged in the EC of a tertiary cancer center rated their fatigue at its worst in the last 24 hours. Patient demographic and clinical factors were retrospectively reviewed from medical records. The chief complaints of patients seeking emergency care included fever, pain, gastrointestinal symptoms, dyspnea, fatigue, and bleeding. More than half (54%) reported severe fatigue (seven or higher on a 0-10 scale) upon EC admission. Moderate to severe pain was highly associated with fatigue severity. Patients with severe fatigue were more likely to be unstable and unable to go home after EC care. In multivariate logistic regression analysis for severe fatigue, the significant risk factors for patients with solid tumors included dizziness (odds ratio [OR]=3.59), severe pain (OR=1.98), poor performance status (OR=1.81), and being female (OR=1.56). Dyspnea was significantly associated with severe fatigue in patients with hematological malignancies (OR=4.74). Although fatigue was not the major reason for an ER visit, single-item fatigue-severity screening demonstrated highly prevalent severe fatigue in sicker EC cancer patients and in those patients who also suffered from other symptoms. PMID- 18411012 TI - Supporting patients and their caregivers after-hours at the end of life: the role of telephone support. AB - Twenty-four hour access is accepted as a gold standard for palliative care service delivery, yet minimal data exist to justify the cost of this initiative to health care planners and policy makers. Further, there is scant information concerning optimal and efficient methods for delivering after-hours palliative care advice and support, particularly in regional and rural settings. This article reports on an evaluation of a local after-hours telephone support service in regional Australia. A centralized after-hours telephone support service was provided by generalist nurses at a Multipurpose Service in a rural community. A mixed-method evaluation, including semistructured interviews, was undertaken after 20 months of operation. During the period March 31, 2005 until November 15, 2006, 357 patients were registered as part of the Mid North Coast Rural Palliative Care Program. Ten percent of patients or their caregivers accessed the After-Hours Telephone Support Service, representing 55 occasions of service. The most common reason for contacting the service was for reassurance surrounding medication usage, symptom management, and anxiety. This experience demonstrates proof of concept that acceptable palliative care advice can be provided by generalist nurses in a cost-efficient manner. Common patterns emerged in utilization that can assist in service planning and staffing formulae. There is also a need to investigate mechanisms of interfacing with larger scale call centers, to explore the differences within generic and disease-specific approaches, and assess the appropriateness of after-hours telephone support with different cultural groups. PMID- 18411013 TI - Fatigue in the general Korean population: application and normative data of the Brief Fatigue Inventory. AB - The purpose of this study was to provide normative data for the Brief Fatigue Inventory (BFI) in the general Korean population so that the results for the general population could be compared with those for patients. We constructed a questionnaire that included the BFI and items on demographic characteristics and conducted a population-based, cross-sectional survey in 1,000 individuals. We used multivariate logistic analysis to investigate factors associated with "usual" and "worst" fatigue. The internal consistency was very high (Cronbach's alpha=0.96) and construct validity was confirmed by factor analysis. All patients had a mean+/-SD BFI score of 4.33+/-2.48 for "worst" fatigue and of 4.07+/-2.27 for "usual" fatigue, and the global BFI score was 3.44+/-2.05. The prevalence of each moderate-to-severe fatigue type was similar in severity of fatigue, with 55.2% in "usual" fatigue, and 57.3% in "worst" fatigue. Among the types of fatigue, the prevalence of severe fatigue was lowest for "usual" fatigue (16.5%). In multivariate analyses, the group aged 40-59 years had greater levels of "usual" and "worst" fatigue compared with the group aged 20-29 years. Poor general health and the presence of comorbidities were also associated with increased "usual" and "worst" fatigue. Regular physical activity was associated with reduced levels of "worst" fatigue. The normal values of BFI with proper psychometric properties may help us to better understand the correlates of fatigue in the general population and patients. Our findings indicate that comorbidities should be considered when comparing fatigue data from the general population with data from patients. PMID- 18411014 TI - Evaluation and management of treatment-related diarrhea in patients with advanced cancer: a review. AB - Diarrhea is a common and significant problem among patients with advanced cancer. Treatment-induced diarrhea can be severe and be associated with life-threatening dehydration and electrolyte abnormalities. The causes of diarrhea among patients with advanced cancer are diverse and some causes of diarrhea require specific therapies. Thus, careful evaluation of the underlying cause is necessary. Palliative care clinicians, particularly those dealing with patients receiving ongoing disease-modifying therapies, must be familiar with the common causes of diarrhea among cancer patients and the strategies to evaluate and manage these common and distressing symptoms. This article addresses four major issues: 1) a review of the causes of treatment-related diarrhea, focusing on diarrhea caused by chemotherapy, targeted therapies, and radiotherapy; 2) differential diagnosis and an approach to evaluation; 3) general management considerations; and 4) cause specific issues in management. PMID- 18411015 TI - Assessing analgesic regimen adherence with the Morisky Medication Adherence Measure for Taiwanese patients with cancer pain. AB - The purposes of this study were (1) to examine the psychometric properties of the Taiwanese version of the Morisky Medication Adherence Measure (MMAM), including its validity and reliability, (2) to investigate levels of analgesic regimen adherence, and (3) to explore the predictors of adherence to the analgesic regimen in a sample of Taiwanese cancer patients with pain. One hundred thirty five patients receiving analgesics for cancer pain participated in this study. Instruments consisted of the Taiwanese version of the MMAM, the Barriers Questionnaire-Taiwan form, the Chinese version of the Brief Pain Inventory, the American Pain Society Outcome Questionnaire, Karnofsky Performance Status, and a demographic questionnaire. Analgesic use ratios were calculated. The Taiwanese version of the MMAM had good psychometric properties for measuring adherence with the analgesic regimens taken by Taiwanese cancer pain patients. Reliability was supported by good internal consistency Cronbach alpha and test-retest coefficients. Validity was corroborated by good known group validity, construct validity, and criterion-related validity. The majority of the patients (51%) showed low levels of medication adherence. The significant predictors for the medication adherence score were age, the Barriers Questionnaire score, and satisfaction with pain management by clinicians after entering pain severity, pain interference with daily life, age, gender, education, types of analgesics used, functional status, and satisfaction with pain management as independent variables. The model accounted for 63% of the variance in the medication adherence score. The Taiwanese version of the MMAM shows excellent reliability and validity. The use of this reliable, valid, simple, and easily administered tool can improve communication between patients and clinicians about use of analgesics and further improve the analgesic regimen adherence. PMID- 18411016 TI - The role of psychological and behavioral variables in quality of life and the experience of bodily pain among persons living with HIV. AB - With increased life expectancy of individuals living with HIV, quality of life (QOL) has become a focus of treatment. More research is needed to address pain related QOL and modifiable variables, such as health behaviors, depressive symptoms, and coping styles, which could be included in treatment protocols to improve QOL among individuals with HIV. Objectives of this study were to (1) examine relationships among health behaviors, psychological variables, and QOL, particularly pain-specific QOL, (2) examine the relationships among coping, depressive symptoms, and QOL, and (3) compare QOL scores of individuals with HIV and population-based normative data. HIV positive men and women not currently on highly active antiretroviral therapy were recruited during regular visits to an HIV outpatient clinic. They completed the Medical Outcome Study Health Survey SF 36 scale, which includes a physical components scale, a mental components scale, and a bodily pain subscale. They also completed questionnaires assessing health behaviors, depressive symptoms, and coping styles. Participants (n=97) scored significantly lower on most aspects of QOL than age-matched Canadian and U.S. norms. Hierarchical multiple regressions revealed that physical activity and CD4 cell count were independently related to lower physical components scale scores; smoking and depressive symptoms were independently associated with lower mental components scale scores; and education, physical activity, and depressive symptoms were independently associated with lower pain-related QOL. Depressive symptoms mediated the relationship between coping styles and the mental components scale and pain-related QOL. Results suggest that targeting depressive symptoms, physical activity, and coping strategies as part of comprehensive treatment protocols could help improve pain-specific QOL and overall QOL among individuals with HIV. PMID- 18411017 TI - Palliative sedation in a specialized unit for acute palliative care in a cancer hospital: comparing patients dying with and without palliative sedation. AB - Palliative sedation is undergoing extensive debate. The aims of this study were to describe the practice of palliative sedation at a specialized acute palliative care unit and to study whether patients who received palliative sedation differed from patients who did not. We performed a systematic retrospective analysis of the medical and nursing records of all 157 cancer patients who died at the acute palliative care unit between 2001 and 2005. Palliative sedation, defined as continuous deep sedation prior to death, was used for 43% of all deceased patients. In 87% of the sedated patients, it was started in the last two days before death. Sedated and nonsedated patients did not differ in survival after admission (eight days vs. seven days, P=0.12). Sedated patients were younger (55 years vs. 59 years, P=0.04) and more often had malignancies of the digestive tract (P<0.01). In both groups, common symptoms at admission were pain (79% vs. 87%, P=0.23), constipation, (40% vs. 48%, P=0.46), and dyspnea (32% vs. 29%, P=0.77). On the day that palliative sedation was started, sedated patients more often suffered from dyspnea and delirium than nonsedated patients at a comparable day before death. The most important indications for palliative sedation were terminal restlessness (60%) and dyspnea (46%). We conclude that at the studied acute palliative care unit, patients who ultimately received palliative sedation did not have symptoms different than nonsedated patients at admission, but on the day at which the sedation was started, they suffered more often from delirium and dyspnea. PMID- 18411018 TI - Intravenous ibandronate rapidly reduces pain, neurochemical indices of central sensitization, tumor burden, and skeletal destruction in a mouse model of bone cancer. AB - Over half of all chronic cancer pain arises from metastases to bone and bone cancer pain is one of the most difficult of all persistent pain states to fully control. Currently, bone pain is treated primarily by opioid-based therapies, which are frequently accompanied by significant unwanted side effects. In an effort to develop nonopioid-based therapies that could rapidly attenuate tumor induced bone pain, we examined the effect of intravenous administration of the bisphosphonate, ibandronate, in a mouse model of bone cancer pain. Following injection and confinement of green fluorescent protein-transfected murine osteolytic 2472 sarcoma cells into the marrow space of the femur of male C3H/HeJ mice, ibandronate was administered either as a single dose (300 microg/kg), at Day 7 post-tumor injection, when tumor-induced bone destruction and pain were first evident, or in three consecutive doses (100 microg/kg/day) at Days 7, 8, and 9 post-tumor injection. Intravenous ibandronate administered once or in three consecutive doses reduced ongoing and movement-evoked bone cancer pain-related behaviors, neurochemical markers of central sensitization, tumor burden, and tumor-induced bone destruction. These results support limited clinical trials that suggest the potential of ibandronate to rapidly attenuate bone pain and illuminate the mechanisms that may be responsible for limiting pain and disease progression. PMID- 18411019 TI - Does palliative care improve quality? A survey of bereaved family members. AB - Palliative care is the interdisciplinary specialty that aims to relieve suffering and improve the quality of care for patients with serious illness and their families. Although palliative care programs are becoming increasingly prevalent in U.S. hospitals, the impact of hospital palliative care consultation programs on the quality of care received by family members is not well understood. We conducted prospective quantitative telephonic interviews of family members of patients who died at Mount Sinai Medical Center between April and December 2005 using the validated "After-Death Bereaved Family Member Interview," to assess quality of medical care at the end of life. Multivariable techniques were used to compare family satisfaction of palliative care patients vs. usual care patients controlling for age, race (white vs. nonwhite), diagnosis (cancer vs. noncancer), socioeconomic status (Medicaid vs. non-Medicaid), and functional status (number of dependent activities of daily living). One hundred ninety eligible subjects were contacted and successful interviews were completed with 149 (78.4%) family members (54 palliative care and 95 usual care patients). Palliative care showed benefit, with 65% of palliative care patients' family members reporting that their emotional or spiritual needs were met, as compared to 35% of usual care patients' family members (P=0.004). Sixty-seven percent of palliative care patients' family members reported confidence in one or more self-efficacy domains, as compared to 44% of usual care patients' family members (P=0.03). Our study shows that palliative care consultation is associated with improved satisfaction, with attention to family and enhanced self-efficacy. Palliative care offers a unique approach by integrating the needs of the family into the care of the patient. PMID- 18411020 TI - Re: subcutaneous methadone-an issue revisited. PMID- 18411021 TI - The concentration-dependent induction of cell death by trimethyltin chloride in rat liver epithelial IAR20 cells. AB - In this study we intended to evaluate the cytotoxic and genotoxic effects of trimethyltin chloride (TMT), one of the most widely used organometallic compounds, on liver cells with the rat liver epithelial cell line IAR20. The results showed that TMT significantly inhibited cell growth in a concentration dependent manner and caused an increase in DNA damage. Additionally, Hoechst 33342 staining and flow cytometry detected increases in apoptotic and necrotic cells. Western blots demonstrated that the Bcl-2 family of proteins was involved in the apoptotic process, but p53 was not concerned. PMID- 18411022 TI - Autoantibody-mediated neuroinflammation: pathogenesis of neuropsychiatric systemic lupus erythematosus in the NZM88 murine model. AB - Autoantibodies play an important role in central nervous system manifestations of neuropsychiatric systemic lupus erythematosus (NPSLE). Previous studies have shown that the lupus-prone NZM88 strain has major neural deficits and high titers of serum IgG to brain antigens. ELISA was performed to detect the presence of IgG in different brain regions of NZM88 mice and to compare the levels with NZM2758 mice and control strains (NZW and BALB/c). There was a substantial increase of IgG in the substantia nigra (SN) and hypothalamus (HT) of brains from NZM88 mice compared to control NZW and BALB/c mice, whereas NZM2758 mice had more IgG in the cortex. The increased presence of IgG in the NPSLE-prone NZM88 mouse brain was paralleled by increased TNF-alpha and IL-12 in the SN and HT regions; significantly elevated expression of MHC Class-II was also observed in the SN of NZM88 mice and cortex of NZM2758 mice. A co-culture system of dopaminergic neurons and microglia was used to demonstrate that NZM88 sera modifies dopaminergic cell activity only in the presence of microglia and that TNF-alpha is synthesized and released in this co-culture. This study demonstrates a functional link between the autoantibodies, the activation of microglia, and neuronal function associated dopamine production, which is suggested to be causally related to the predominant NPSLE syndromes. PMID- 18411023 TI - The accumulative effects of modifiable risk factors on inflammation and haemostasis. AB - Various modifiable risk factors have been associated with inflammation and haemostasis, although the accumulative effects have not yet been examined. We therefore explored additive and independent associations of modifiable risk factors (smoking, alcohol, cholesterol, obesity, hypertension, physical activity) with inflammatory (CRP) and haemostatic (fibrinogen) markers. Data were collected from a sample of 7670 healthy asymptomatic participants (45.9% men, aged 46.2+/ 15.6 years). A graded increase in the risk of inflammation (CRP> or =3 mg/L) with increasing numbers of modifiable risk factors was demonstrated (odds ratio for > or =4 risk factors=5.09, 95% CI, 3.96-6.55). Similar associations were found in relation to haemostasis. Central adiposity was the strongest independent predictor of inflammation (OR=3.45, 95% CI, 3.07-3.87) although smoking most strongly predicted haemostasis (OR=2.19, 95% CI, 1.94-2.48). These findings suggest that targeting multiple risk factors is likely to have the greatest benefit for cardiovascular prevention. PMID- 18411024 TI - Outcome after thoracic aortic injury: experience in a level-1 trauma center. AB - Traumatic rupture of the thoracic aorta is a potentially fatal injury that leads to death in 75-90% of cases at the time of injury. In this report, we present our experience with traumatic thoracic aortic injury and compare the outcome in patients with respect to their hemodynamic stability at presentation and the timing of surgical repair. We performed a retrospective data analysis of the medical records of 30 patients who had sustained a traumatic rupture of the thoracic aorta during the period from January 1, 2000 to October 30, 2005. The demographic data, mechanism of injury, modality of diagnosis, location of injury, other associated injuries, hemodynamic stability at presentation, response to resuscitation, timing of aortic repair, as well as the resultant morbidities and mortalities were reviewed. Traumatic rupture of the thoracic aorta was diagnosed in 30 patients. The injury was located in the ascending aorta in two patients, in the aortic isthmus in 25 patients, and in the descending aorta (distal to the isthmus) in three patients. Associated injuries included head injury (50%), C spine (23.3%), lung injury (80%), and visceral (63%) and extremity (60%) injury. Seven patients (23%) were pronounced dead on arrival to the emergency room, 14 patients (47%) were hemodynamically unstable upon arrival, and nine patients (30%) were hemodynamically stable. In the unstable group, two patients (14%) expired before operative repair, 11 patients (79%) underwent emergent repair of the thoracic aorta resulting in 46% mortality, and one patient (7%) underwent delayed repair after initial stabilization with a splenectomy for a splenic laceration. In the stable group, three patients (33%) underwent early (within 24 hr) aortic surgery, while repair was delayed in six (67%). The mean duration of time spent prior to delayed repair was 20.85 days (range 2-53, median = 25). There was no mortality in this group. Acute traumatic thoracic aortic rupture remains a highly fatal condition. Hemodynamic instability remains the main mortality risk factor. Delayed repair is safe and is not associated with increased risk of aortic rupture in hemodynamically stable patients. PMID- 18411025 TI - Arterial thrombosis after intravenous infusion of oral bacterium in a rat model. AB - Oral bacteria have been detected at atherosclerotic plaque, aneurysms, and thrombosed arteries in Buerger disease. We explored a possible relationship between the oral bacterium Porphyromonas gingivalis and arterial thrombosis at proximal and distal sites in rats. Eighteen rats underwent subcutaneous placement of an infusion pump connected to the jugular vein. The Pg infusion group received a continuous infusion of P. gingivalis for 2 weeks, and the controls received normal saline. At 2 and 4 weeks, specimens were obtained from the iliac, superficial, and below-knee arteries, which were studied pathologically and by polymerase chain reaction (PCR) analysis to detect P. gingivalis-specific DNA. The Pg infusion group had thrombosis in 33.3% at 2 weeks and in 55.6% at 4 weeks, but normal arterial wall structure was preserved without any features of infection. Positive PCR findings were recognized in 73.3% and 22.2% at 2 and 4 weeks, respectively. At 4 weeks, thrombosis was observed in a higher proportion, with the below-knee specimens having an especially high thrombus rate (83.3%). No control specimen had thrombosis or positive PCR results. Bacteremia due to the oral pathogen P. gingivalis may lead to thrombus formation in the peripheral arteries, especially in small-sized arteries. PMID- 18411026 TI - Long-term outcome following stent reconstruction of the aortic bifurcation and the role of geometric determinants. AB - We assessed the long-term patency of kissing stent reconstruction of the aortoiliac bifurcation and identified variables that may influence it. We retrospectively reviewed our experience with stent-reconstruction procedures of the aortoiliac bifurcation from January 1998 through June 2005. The impact of demographic variables, vascular risk factors, disease location and characteristics, stent material and design, and stenting configuration on stent patency was assessed using univariate and multivariate analysis. In particular, we evaluated the effect of geometric mismatch between the protruding segment of the stents and the distal aortic lumen. Sixty-six patients underwent aortobi iliac stent reconstruction. Indications were bifurcation or bilateral proximal iliac disease in 52 patients and unilateral ostial disease requiring contralateral protection in 14 patients. Limited disease (TASC A and B) was present in 40 limbs in 19 patients; extensive/diffuse disease (TASC C and D) was present in 78 limbs in 47 patients. Complete occlusions were present in 37 limbs in 28 patients (bilateral in nine patients). Self-expanding stents were used in 56 procedures and balloon-expandable stents in 10. Crossing configuration was used in 43 procedures, while abutting configuration was used in 23 procedures. Technical success was achieved in 62 patients (94%), with all four failures due to inability to cross a chronically occluded limb. Three of these patients underwent aortomono-iliac stenting with a crossover femoral-femoral bypass graft, with the remaining one opting for no further interventions. Median combined follow-up was 37 +/- 27 months (range 0-102). Hemodynamically significant restenosis developed in nine patients (14%). The management of restenosis was endovascular in eight patients and was successful in all (balloon dilation in four, restenting in three, thrombolysis and stenting in one) and operative in one patient who developed aortic occlusion and underwent aortobifemoral grafting. Survival table analysis showed primary and assisted patency rates at 4 years of 81% and 94%, respectively. The mortality rate during follow-up was 19 (cardiac cause in eight, pulmonary cause in three, and malignancy in five). Univariate analysis showed radial mismatch (aortic lumen dead space around the protruding segment of the stents), female gender, prior occlusion, and residual stenosis to be significant predictors of restenosis. Multivariate logistic regression analysis showed radial mismatch to be the only significant determinant of restenosis, although the statistical power of the model was limited by the small number of restenoses. Stent reconstruction of the aortoiliac bifurcation for occlusive disease is effective and durable, even with complex aortoiliac disease and long segment occlusions. Most restenoses are amenable to endovascular treatment, with excellent long-term assisted patency. Geometric variables related to individual aortic anatomy and disease pattern (patient-dependent) and stenting configuration (operator-dependent) may have an impact on long-term patency. PMID- 18411027 TI - Does diabetes mellitus play a role in restenosis and patency rates following lower extremity peripheral arterial revascularization? A critical overview. AB - Patients with diabetes mellitus (DM) are at increased risk of developing lower extremity peripheral arterial disease (PAD). The effect of DM on restenosis and patency rates in patients with PAD undergoing surgical revascularization or percutaneous interventions has not been fully clarified. We therefore critically reviewed the role of DM in restenosis, as well as primary and secondary patency rates in these patients. We searched Medline for studies investigating the effect of DM on restenosis (primary and secondary patency) rates in patients undergoing surgical/percutaneous interventions for the treatment of lower extremity PAD. Search terms used were "diabetes and peripheral arterial disease," "angioplasty," "restenosis," "revascularization," "patency rates," and "in-stent restenosis." Diabetic patients with PAD have similar restenosis, primary patency, and secondary patency rates compared with nondiabetic patients. However, mortality and amputation rates are increased in patients with DM. This increased risk of mortality and amputation may distort the estimation of restenosis and patency rates. Strict glucose control should be implemented in diabetic patients. Additionally, the use of antiplatelet agents and statins may have a beneficial effect on restenosis and patency rates. The role of radiation therapy in preventing restenosis remains to be determined. Patients with PAD and DM should receive optimal medical therapy to improve cardiovascular outcome and decrease functional decline. The direct involvement of vascular surgeons in the management of PAD patients is essential to reduce the incidence of cardiovascular events and mortality rates. PMID- 18411028 TI - Outcomes of endoluminal therapy for ostial disease of the major branches of the aortic arch. AB - While aggressive endoluminal therapy for occlusive disease of the major branches of the arch of the aorta (brachiocephalic [BCA], left common carotid [LCCA], and left subclavian [LSCA] arteries) is commonplace, long-term outcomes in this population are unclear. We examined the long-term outcomes of endoluminal therapy for ostial aortic arch disease at a single tertiary referral academic medical center. A prospective database of patients undergoing endovascular treatment of aortic arch vessel atherosclerotic occlusive disease between 1990 and 2004 was maintained and retrospectively analyzed. Patients with stenotic ostial lesions of the major thoracic aorta branches were selected. Angiograms were reviewed in all cases to assess lesion characteristics. Patency was assessed by routine clinical and, in the LCCA and LSCA, duplex ultrasound follow-up at 1, 6, and 12 months postintervention and every 12 months thereafter. Results were standardized to current Trans-Atlantic Inter-Society Consensus and Society for Vascular Surgery criteria. Kaplan-Meier analyses were performed to assess time-dependent outcomes. Factor analyses were performed using a Cox proportional hazard model for time dependent variables. Data are presented as mean +/- SEM. Forty-four patients (average age 64 +/- 2 years, 59% male) underwent 26 LSCA, 11 LCCA, and eight BCA interventions for primary indications of arm ischemia (29%), prevention or treatment of coronary steal syndrome (29%), or cerebrovascular signs/symptoms (42%). The technical success rate was 98%, with a 90-day mortality rate of 0% and a major adverse event rate of 2%. There were no strokes and no upper extremity embolic events. Cumulative patency was 88 +/- 8% at 3 years, with a reintervention rate of 7%. The overall symptom recurrence rate was 4%. No local or systemic factors were associated with poor outcomes. Endoluminal stenting for ostial disease of the branches of the aortic arch provides excellent and long term patency rates with low morbidity, mortality, and secondary intervention rates. With an overall technical success of 98%, our results parallel those for lesions located more distally in the arch branches and support the continued use of percutaneous therapy for atherosclerotic disease throughout the arch branches. PMID- 18411029 TI - Measure what matters: institutional outcome data are superior to the use of surrogate markers to define "center of excellence" for abdominal aortic aneurysm repair. AB - Outcome analysis is increasingly being used to develop health-care policy and direct patient referral. For example, the Leapfrog Group health-care quality initiative has proposed "evidence-based hospital" referral criteria for specific procedures including elective abdominal aortic aneurysm repair (AAA-R). These criteria include an annual hospital AAA operative volume exceeding 50 cases and provision of intensive care unit (ICU) care by board-certified intensivists. Outcomes after AAA-R are reportedly influenced by presentation (intact vs. ruptured), operative approach (endovascular vs. open, transperitoneal vs. retroperitoneal), surgeon subspecialty, case volume (hospital and surgeon), and provision of postoperative care by an intensivist. The purpose of this study was to compare our single-center results with those of high-volume centers to assess the validity of the concept that surrogate markers, such as case volume or intensivist involvement, can be used to estimate procedural outcome. A retrospective review was performed of AAA-Rs at one low-volume academic medical center from January 1994 to March 2005. Demographic data, aneurysm diameter and location, operative indications, and repair approach were documented. Postoperative complications, mortality rates, and hospital and ICU length of stay (LOS) were noted and compared to established benchmarks. During the study period, 270 patients underwent AAA-R (annual mean = 27 hospital cases and 13.4 cases/attending vascular surgeon). ICU care was provided by a dedicated vascular surgery service without routine intensivist involvement. Open, elective, infrarenal AAA-R was performed in 161 patients (60%), with a 2.5% hospital mortality rate (30-day, 3.1%). Thirty-three (12%) patients underwent elective endovascular aneurysm repair (EVAR), with no mortality. Both ICU (3.7 vs. 1.4 days, p = 0.03) and hospital (9.2 vs. 2.8 days, p = 0.002) LOS were significantly reduced after EVAR compared to open repair. Hospital LOS was significantly lower after open retroperitoneal repair compared to transperitoneal repair (6.1 vs. 10.3 days, p = 0.001). Thirty-five patients (13%) underwent ruptured AAA-R, with only 34.3% mortality (in-hospital and 30-day). Forty-one patients (15%) underwent repair of complex aortic aneurysms, with 14.1% mortality. There are increasing societal and economic pressures to direct patient referrals to "centers of excellence" for specific surgical procedures. Although our institution meets neither of the Leapfrog Group's proposed criteria, our mortality and LOS for both intact and ruptured infrarenal AAA-R are equivalent or superior to published benchmarks for high-volume hospitals. Individual institutional outcome results such as these suggest that patient referral and care should be based upon actual, carefully verified outcome data rather than utilization of surrogate markers such as case volume and subspecialist involvement in postoperative care. PMID- 18411030 TI - Total robotic ligation of inferior mesenteric artery for type II endoleak after endovascular aneurysm repair. AB - We present a case of totally robotic ligation of the inferior mesenteric artery (IMA) for treatment of a persistent endoleak from the IMA into the aneurysm sac after endovascular aneurysm repair (EVAR). An 84-year-old male underwent EVAR with a Gore Excluder stent graft for an asymptomatic infrarenal abdominal aortic aneurysm. Follow-up computed tomographic (CT) scan showed persistent type II endoleak from the IMA, with progressive enlargement of the aneurysm sac from 5 to 6.1 cm over an 18-month period. In this case, the patient underwent ligation of the IMA using the da Vinci Surgical System for the treatment of retrograde flow into the aneurysm sac. The total operating time was 249 min; of this, the robotic assistance time was approximately 180 min. No intraoperative complications occurred. The estimated blood loss was 50 mL and the urine output 650 mL. The patient was extubated immediately after the procedure and tolerated a regular diet the following day. He was discharged home with a urinary catheter on postoperative day 2. CT scan postoperatively and at 3-month follow-up demonstrated an occluded IMA and stabilization of the aneurysm sac size. PMID- 18411031 TI - Follow-up of aneurysm neck diameter after endovascular repair of abdominal aortic aneurysms. AB - We assessed aneurysm neck diameter change after endovascular abdominal aortic aneurysm repair (EVAR) and its relationship to stent-graft diameter. Ninety-eight patients with abdominal aortic aneurysm who underwent EVAR were studied with multislice computed tomography following a standardized protocol. A preoperative study and immediate postoperative, 6-month, 1-year, and 2-year follow-up studies were performed. The aneurysm neck was measured from adventitia to adventitia, 6 mm below the lowermost renal artery, in planar images performed perpendicular to the vessel axis (real axial section). Baseline and follow-up neck diameters were compared with stent-graft diameters. For statistical analysis, a one-way analysis of variance with repeated measures was used. Pearson's correlation coefficient was used to examine the correlation between the change in neck diameter and stent graft diameter. The average neck diameter was 22.38 mm (range 16-32.5) on the preoperative study and 23.35 mm (17-33.9) on the immediate postoperative, 24.35 mm (18.2-34.5) on the 6-month, 24.36 mm (18-34.5) on the 1-year, and 24.39 mm (17.8-35.7) on the 2-year follow-up. The mean device diameter was 24.08 mm (20 32). A significant increase in average neck diameter was found between the preoperative, immediate postoperative control, and 6-month control. There was no significant increase in the average neck diameter between the 6-month, 1-year, and 2-year follow-up. Baseline mean stent-graft oversizing was 1.7 mm, which decreased to -0.31 mm at latest follow-up. Dilation of the neck did not significantly exceed the endograft diameter in 83 cases (87.36%). An enlargement of the infrarenal aneurysm neck occurred during the first 6 months after EVAR. No significant variation in neck diameter occurred between the 6-month and 2-year follow-up visits. In the majority of cases, dilation of the aneurysm neck does not significantly exceed stent-graft diameter and, therefore, is possibly related to the presence of the endograft. PMID- 18411032 TI - Endovascular repair of a chronic aortocaval fistula using a thoracic aortic endoprosthesis. AB - Penetrating abdominal trauma with injury to the aorta and vena cava usually requires emergent intervention and is frequently lethal. Formation of a chronic aortocaval fistula (ACF) is an uncommon late complication of these injuries. We report a case of an ACF presenting 17 years after a gunshot wound to the abdomen, with progressive congestive heart failure as the presenting symptom. The ACF was successfully treated with an endoprosthesis designed for the thoracic aorta. PMID- 18411033 TI - Midterm patency following atherectomy for infrainguinal occlusive disease: a word of caution. AB - There has been widespread initial enthusiasm for peripheral atherectomy using the SilverHawk device. We sought to evaluate our midterm patency following infrainguinal atherectomy. Nineteen consecutive patients underwent 23 separate atherectomy procedures on 20 limbs from March 2005 through June 2006 (11 males, age 66 +/- 14 years). The primary lesions were atherosclerotic (n = 18) and vein graft stenoses (n = 2). Three additional procedures were redo atherectomies for restenotic lesions. The TASC classification of the primary lesions was A in 3, B in 9, and C in 8. The median number of treated lesions per limb was 2 (range 1 4). The location of the most distal native vessel stenosis was the superficial femoral artery in 12, popliteal artery in six, and crural artery in two. Atherectomy was successful in 18 primary procedures and all three repeat atherectomy procedures. Touch-up balloon dilatation was used in five procedures. Complications included one groin hematoma and two perforations, treated with stenting in one and bypass grafting in one. Preoperative ankle-brachial index and transmetatarsal pulse volume recording were 0.51 +/- 0.16 and 3.3 +/- 0.8, respectively, which at 1-month improved to 0.80 +/- 0.16 and 2.4 +/- 0.4 (p < 0.001). Only two vessels remained patent at 12 months. Recurrence developed in 16 of the successful primary procedures, including both vein graft lesions and all three repeat atherectomy procedures. The mode of recurrence was restenosis in 14 and occlusion/thrombosis in five. Secondary interventions included balloon angioplasty/thrombolysis in two, stenting in three, redo atherectomy in three, vein bypass grafting in five, and observation alone in one. Major limb amputation was required in five patients. Primary patency rates per treated limb at 3, 6, and 12 months were 38%, 10%, and 10%. The corresponding assisted patency rates were 50%, 23%, and 10%. Our experience suggests a very poor midterm patency of excisonal atherectomy using the SilverHawk device, although a 74% limb salvage rate was maintained through secondary interventions. Liberal use of this technology is associated with high cost and frequent requirement of reintervention. PMID- 18411034 TI - The performance of the time-frequency analysis software (TF32) in the acoustic analysis of the synthesized pathological voice. AB - The purpose of this study was to examine the algorithm-measuring capabilities used in the Time-Frequency Analysis Software Program for 32-bit Windows (TF32) for measuring fundamental frequency (F0), its dependent measures, and signal-to noise ratio (SNR). The stability, accuracy, and linearity of its algorithm to systematic changes in aspiration noise and/or spectral slope (to mimic the perceptual characteristics of breathiness, roughness, and hoarseness) were evaluated using its analysis output to five female and five male synthesized voices. TF32 was used to calculate F0, Jitter%, Shimmer%, and SNR for each of the synthesized signals. The findings indicate that although TF32 produced stable results for male synthesized samples, they were not accurate when measuring F0, Jitter%, and Shimmer% with the addition of noise and variations in open quotient independently and in combination. In contrast, TF32 was neither stable nor accurate in making the same measurements for female synthesized samples. However, TF32 was stable and accurate in measuring SNR for male and most of female voices. These results point to an inappropriate F0 extraction algorithm in TF32 and stress the need for further research to remediate the algorithm or to identify a superior one. PMID- 18411035 TI - Resonant voice in acting students: perceptual and acoustic correlates of the trained Y-Buzz by Lessac. AB - This study aimed to investigate perceptually and acoustically Lessac's Y-Buzz and sustained productions of Brazilian Portuguese habitual /i/ vowels pre- and posttraining and to verify the presence of formant tuning and its association with the perception of a more resonant voice. The subjects of this study were 54 acting students, 31 female and 23 male, with no voice problems, distributed in seven groups. Each group received four weekly sessions of training. For the pretraining recording, they were asked to sustain the vowel /i/ in a habitual mode three times at self-selected comfortable frequencies and intensity. After training, they repeated the habitual /i/ and also the trained Y-Buzz. Five voice specialists rated how resonant each sample sounded. The fundamental frequency (F(0)), the first four formant frequencies, the distance between the frequencies of F(1) and F(0) were measured, as well as the harmonic frequency (H(2)) frequency and the difference between F(1) and H(2) in the case of male voices (Praat 4.4.33, Institute of Phonetic Sciences, University of Amsterdam, The Netherlands). The trained Y-Buzz was considered more resonant than the habitual /i/ samples, regardless the gender and demonstrated a lowering of the four formant frequencies. F(1) was especially lower in both groups (288Hz-female and 285Hz-male), statistically significant in the female group. The F(1)-F(0) difference was significantly smaller for the female Y-Buzz (52Hz), as well as F(1)-H(2) in the case of the male Y-Buzz (12Hz), suggesting formant tuning. It was not possible to establish association between the perceptual grades and measures F(1)-F(0) or F(1)-H(2). PMID- 18411036 TI - Acoustic analysis of aperiodic voice: perturbation and nonlinear dynamic properties in esophageal phonation. AB - Esophageal voice is a method of voice production after total laryngectomy. Previous research suggests that perturbation analysis may inaccurately measure aperiodic voices and that nonlinear dynamic methods may be more appropriate for analyzing signals of this type. Therefore, we hypothesized that nonlinear dynamic analysis would be more capable than perturbation parameters for reliable measurement of the aperiodic esophageal voice. The study design was acoustic comparison of esophageal and normal voice cohorts using nonlinear dynamic and perturbation measures. Twenty subjects in two age-matched groups participated in the study. Jitter, shimmer, signal-to-noise ratio (SNR), correlation dimension, and second-order entropy were measured from audio recordings of subjects' voices. Jitter and shimmer values were significantly higher and SNR values were significantly lower for esophageal voices than for normal voices. Error (err) count values, which indicate perturbation analysis reliability, were 0 in normal voices and significantly higher in esophageal voices. Error was attributable to signal aperiodicity and demonstrated that perturbation analysis yielded questionable results for esophageal voice. However, nonlinear dynamics measures analyzed both cohorts reliably and indicated that esophageal voice was significantly more chaotic than normal voice. The results demonstrated the capability of nonlinear dynamic methods to reliably quantify both aperiodic and periodic signals and differentiate normal from esophageal voices. It is suggested that nonlinear dynamic analysis be used preferentially for acoustic characterization of aperiodic voices, such as esophageal voice. Future research should focus on clarification of perturbation analysis reliability and further application of nonlinear dynamic measures to aperiodic voices. PMID- 18411037 TI - The effect of specific versus combined warm-up strategies on the voice. AB - Although actors and singers typically warm-up the voice before performing, little is known about the effects of vocal warm-up on the voice. This study was designed to determine the relative effectiveness of specific versus combined warm-up strategies on the voice by group comparison. Twenty participants, 10 male and 10 female actors, completed two warm-up protocols, at least 1 week apart, in a counter-balanced order. Measures of phonation threshold pressure (PTP), jitter, noise-to-harmonics-ratio (NHR), and self-perceived vocal effort were obtained. For the males, there was no significant difference in PTP difference values between vocal warm-up only (specific) and vocal plus aerobic warm-up (combined) conditions. For the females, however, a significant difference was found, with a greater reduction in PTP in the combined warm-up condition. A significant difference was also found in male jitter percent values, with significantly lower values in the specific condition than in the combined condition. There were no significant differences in the remaining comparisons. Females appeared to respond more to the vocal warm-up when it had an aerobic exercise component. It is possible that the males' greater level of physical fitness impacted this finding. PMID- 18411038 TI - Individual allocation had an advantage over cluster randomization in statistical efficiency in some circumstances. AB - OBJECTIVE: To investigate the trade off between performing an individual randomized trial with a Complier Average Causal Effect analysis and accepting the fact that there will be some contamination, with a cluster randomized trial and the subsequent effect on the sample size and power of the trial. STUDY DESIGN AND SETTING: Monte Carlo simulations were undertaken to generate trial data where there was some contamination of control participants. The trials were simulated so the null hypothesis was false. Assessments were made of whether a type II error had been committed (i.e., whether the false null had not been rejected). RESULTS: As contamination increases, the power of the study to detect a true difference between the two groups declines. Using a Complier Average Causal Effect approach, unless anticipated contamination exceeds 30%, retains a sample size advantage over the cluster randomized design despite a relatively small cluster (i.e., 10) and reasonably small ICC (i.e., 0.04). CONCLUSION: If contamination can be measured precisely or estimated then in some circumstances individual allocation has an advantage over cluster randomization in statistical efficiency. PMID- 18411039 TI - Reporting of adverse events in systematic reviews can be improved: survey results. AB - OBJECTIVE: To assess how information about adverse events is included in systematic reviews. STUDY DESIGN AND SETTING: We included all new Cochrane reviews published in the Cochrane Database of Systematic Reviews (CDSRs) and all new reviews (2003--2004) in the Database of Abstracts of Reviews of Effects (DAREs) in Issue 1 2005 of The Cochrane Library. RESULTS: More than half of Cochrane (44/78) and DARE (46/79) reviews assessed drug interventions. The rest assessed surgery (Cochrane [12]; DARE [10]), psychosocial, educational, or physiotherapy interventions (22; 23). Seventy-six percent (59/78) of Cochrane reviews mentioned adverse events as an outcome compared with 48% (38/79) of DARE reviews. Most reviews mentioning adverse events were of drug interventions (Cochrane [41/59]; DARE reviews [29/38]). Considering reviews that mentioned adverse events, 95% (56/59) of Cochrane reviews included only randomized trials and 73% (43/59) included an analysis of adverse events. For 10 Cochrane reviews, adverse events had not been reported by the included trials. In contrast, 58% (22/38) of DARE reviews mentioning adverse events included only randomized trials, the rest included both randomized and nonrandomized studies. CONCLUSIONS: Most Cochrane reviews of drug interventions considered adverse events. This was not the case for DARE reviews and for Cochrane reviews of nondrug interventions. This could be improved. PMID- 18411040 TI - Trial sequential analysis reveals insufficient information size and potentially false positive results in many meta-analyses. AB - OBJECTIVES: To evaluate meta-analyses with trial sequential analysis (TSA). TSA adjusts for random error risk and provides the required number of participants (information size) in a meta-analysis. Meta-analyses not reaching information size are analyzed with trial sequential monitoring boundaries analogous to interim monitoring boundaries in a single trial. STUDY DESIGN AND SETTING: We applied TSA on meta-analyses performed in Cochrane Neonatal reviews. We calculated information sizes and monitoring boundaries with three different anticipated intervention effects of 30% relative risk reduction (TSA(30%)), 15% (TSA(15%)), or a risk reduction suggested by low-bias risk trials of the meta analysis corrected for heterogeneity (TSA(LBHIS)). RESULTS: A total of 174 meta analyses were eligible; 79 out of 174 (45%) meta-analyses were statistically significant (P<0.05). In the significant meta-analyses, TSA(30%) showed firm evidence in 61%. TSA(15%) and TSA(LBHIS) found firm evidence in 33% and 73%, respectively. The remaining significant meta-analyses had potentially spurious evidence of effect. In the 95 statistically nonsignificant (P>or=0.05) meta analyses, TSA(30%) showed absence of evidence in 80% (insufficient information size). TSA(15%) and TSA(LBHIS) found that 95% and 91% had absence of evidence. The remaining nonsignificant meta-analyses had evidence of lack of effect. CONCLUSION: TSA reveals insufficient information size and potentially false positive results in many meta-analyses. PMID- 18411041 TI - The reporting quality of meta-analyses improves: a random sampling study. AB - OBJECTIVES: To determine the overall quality of reporting of meta-analyses (QUOROMs) of randomized controlled trials (RCTs) based on the QUOROM statement, to compare the reporting quality of paper-based articles and Cochrane reviews, and to determine whether compliance with the statement improves over time. STUDY DESIGN AND SETTING: A random sample of systematic reviews or meta-analyses of RCTs was selected from Medline (2000-2005). RESULTS: A total of 161 articles were included. The mean QUOROM score was 12.3 (95% confidence interval [CI]: 12.0, 12.6), which rose from 10.5 (95% CI: 8.8, 12.1) in 2000 to 13.0 (95% CI: 12.2, 13.8) in 2005. The mean QUOROM scores of Cochrane reviews and paper-based articles were 14.2 (95% CI: 13.9, 14.5) and 11.7 (95% CI: 11.3, 12.1), respectively. Compared with the paper-based articles, the Cochrane reviews had better reporting quality in the abstract section, while the quality of their trial flows was poor. The fulfillment of most QUOROM items improved with time. A linear relation of the QUOROM score with time was revealed. CONCLUSIONS: The reporting quality of meta-analyses improves with time. The reporting quality of Cochrane reviews is better compared with paper-based articles. Room still exists for improvements in the reporting quality of both Cochrane and paper-based articles. PMID- 18411042 TI - Cardiologists' charting varied by risk factor, and was often discordant with patient report. AB - OBJECTIVE: To assess the completeness of cardiac risk factor documentation by cardiologists, and agreement with patient report. STUDY DESIGN AND SETTING: A total of 68 Ontario cardiologists and 789 of their ambulatory cardiology patients were randomly selected. Cardiac risk factor data were systematically extracted from medical charts, and a survey was mailed to participants to assess risk factor concordance. RESULTS: With regard to completeness of risk factor documentation, 90.4% of charts contained a report of hypertension, 87.2% of diabetes, 80.5% of dyslipidemia, 78.6% of smoking behavior, 73.0% of other comorbidities, 48.7% of family history of heart disease, and 45.9% of body mass index or obesity. Using Cohen's k, there was a concordance of 87.7% between physician charts and patient self-report of diabetes, 69.5% for obesity, 56.8% for smoking status, 49% for hypertension, and 48.4% for family history. CONCLUSION: Two of four major cardiac risk factors (hypertension and diabetes) were recorded in 90% of patient records; however, arguably the most important reversible risk factors for cardiac disease (dyslipidemia and smoking) were only reported 80% of the time. The results suggest that physician chart report may not be the criterion standard for quality assessment in cardiac risk factor reporting. PMID- 18411043 TI - HER2-dependent MMP-7 expression is mediated by activated STAT3. AB - MMP-7 expression is highly regulated at the level of transcription. An understanding of how the MMP-7 gene is regulated is critical to elucidate the mechanisms of MMP-7 overexpression in the early tumor development. In the present study, increased mRNA and protein expressions of MMP-7 were observed in MCF-7 cells stably overexpressing HER2 (MCF-7/HER2). The promoter activity of MMP-7 gene was upregulated in MCF-7/HER2 cells and significantly enhanced by HRG induction. Examination of the MMP-7 promoter sequence revealed three potential STAT3 binding sites within the proximal region. MMP-7 promoter activity was remarkably induced in MCF-7 cells expressing the constitutively activated STAT3 (MCF-7/STAT3C). RT-PCR and Western blot analysis confirmed the expression upregulation of mRNA and protein of MMP-7 in the MCF-7/STAT3C cells. Binding of STAT3 to MMP-7 promoter was verified by ChIP and the critical STAT3 element within the MMP-7 promoter identified by the mutagenesis of the core STAT3 recognition sequence. Increased STAT3 phosphorylation was observed in either HER2 overexpressing cells or HRG-induced cells. The data indicate that HRG-induced HER2-dependent transcriptional upregulation and protein secretion of MMP-7 are mediated by activated STAT3. The expression of MMP-7 may be attributed to HER2/STAT3 activation. PMID- 18411044 TI - Cutaneous concentration of lycopene correlates significantly with the roughness of the skin. AB - Antioxidant substances in the skin are expected to slow down photo ageing. We therefore developed the hypothesis that high levels of antioxidant substances may be correlated to lower levels of skin roughness. By utilizing modern optical non invasive in vivo methods, the structures of the furrows and wrinkles as well as the concentration of lycopene were analyzed quantitatively on the forehead skin of 20 volunteers aged between 40 and 50 years. In a first step, the age of the volunteers was correlated to their skin roughness. Here, no significant correlation was found. In a second step, a significant correlation was obtained between the skin roughness and the lycopene concentration (R=0.843). These findings indicate that higher levels of antioxidants in the skin effectively lead to lower levels of skin roughness, and therefore support our hypothesis. PMID- 18411045 TI - Influence of the delivery systems using a microneedle array on the permeation of a hydrophilic molecule, calcein. AB - Despite the advantages of drug delivery through the skin, such as easy accessibility, convenience, prolonged therapy, avoidance of the liver first-pass metabolism and a large surface area, transdermal drug delivery is only used with a small subset of drugs because most compounds cannot cross the skin at therapeutically useful rates. Recently, a new concept was introduced known as microneedles and these could be pierced to effectively deliver drugs using micron sized needles in a minimally invasive and painless manner. In this study, biocompatible polycarbonate (PC) microneedle arrays with various depths (200 and 500 microm) and densities (45, 99 and 154 ea/cm2) were fabricated using a micro mechanical process. The skin permeability of a hydrophilic molecule, calcein (622.5D), was examined according to the delivery systems of microneedle, drug loading, depth of the PC microneedle, and density of the PC microneedle. The skin permeability of calcein was the highest when the calcein gel was applied to the skin with the 500 microm-depth PC microneedle, simultaneously. In addition, the skin permeability of calcein was the highest when 0.1g of calcein gel was coupled to the 500 microm-depth PC microneedle (154 ea/cm2) as well as longer microneedles and larger density of microneedles. Taken together, this study suggests that a biocompatible PC microneedle might be a suitable tool for transdermal drug delivery system of hydrophilic molecules with the possible applications to macromolecules such as proteins and peptides. PMID- 18411047 TI - Mono-component versus binary isotherm models for Cu(II) and Pb(II) sorption from binary metal solution by the green alga Pithophora oedogonia. AB - The sorption of Cu(II) and Pb(II) by Pithophora markedly decreased as the concentration of the secondary metal ion, Cu(II) or Pb(II), increased in the binary metal solution. However, the test alga showed a greater affinity to sorb Cu(II) than Pb(II) from the binary metal solution. Mono-component Freundlich, Langmuir, Redlich-Peterson and Sips isotherms successfully predicted the sorption of Cu(II) and Pb(II) from both single and binary metal solutions. None of the tested binary sorption isotherms could realistically predict Cu(II) and Pb(II) sorption capacity and affinity of the test alga for the binary metal solutions of varying composition, which mono-component isotherms could very well accomplish. Hence, mono-component isotherm modeling at different concentrations of the secondary metal ion seems to be a better option than binary isotherms for metal sorption from binary metal solution. PMID- 18411048 TI - Tubulysin analogs incorporating desmethyl and dimethyl tubuphenylalanine derivatives. AB - A series of tubulysin analogs in which one of the stereogenic centers of tubuphenylalanine was eliminated were synthesized. All compounds were tested for antiproliferative activity towards ovarian cancer cells and for inhibition of tubulin polymerization. The dimethyl analogs were generally more active than the desmethyl analogs, and four analogs have tubulin polymerization IC(50) values similar to combretastatin A4 and the hemiasterlin analog HTI-286. PMID- 18411046 TI - Fibroblast-derived 3D matrix differentially regulates the growth and drug responsiveness of human cancer cells. AB - Recent studies have emphasized the importance of cellular microenvironment in modulating cell growth and signaling. In vitro, collagen matrices, Matrigel, and other synthetic support systems have been used to simulate in vivo microenvironments, and epithelial cells grown in these matrices manifest significant differences in proliferation, differentiation, response to drugs, and other parameters. However, these substrates do not closely resemble the mesenchymal microenvironment that is typically associated with advanced carcinomas in vivo, which is produced to a large extent by fibroblasts. In this study, we have evaluated the ability of a fibroblast-derived three-dimensional matrix to regulate the growth of a panel of 11 human tumor epithelial cell lines. Although proliferative and morphological responses to three-dimensional cues segregated independently, general responsiveness to the matrix correlated with the ability of matrix to influence drug responses. Fibroblast-derived three dimensional matrix increased beta1-integrin-dependent survival of a subset of human cancer cell lines during taxol treatment, while it sensitized or minimally influenced survival of other cells. beta1-integrin-dependent changes in cell resistance to taxol did not correlate with the degree of modulation of FAK and Akt, implying that additional signaling factors are involved. Based on these results, we propose that these matrices potentially have value as in vitro drug screening platforms. PMID- 18411049 TI - Antagonists of the human adenosine A2A receptor. Part 3: Design and synthesis of pyrazolo[3,4-d]pyrimidines, pyrrolo[2,3-d]pyrimidines and 6-arylpurines. AB - A series of pyrazolo[3,4-d]pyrimidine, pyrrolo[2,3-d]pyrimidine and 6-arylpurine adenosine A(2A) antagonists is described. Many examples were highly selective against the human A(1) receptor sub-type and were active in an in vivo model of Parkinson's disease. PMID- 18411050 TI - Prevalence of metabolic syndrome and serum marker levels in patients with four subtypes of cerebral infarction in Japan. AB - In this hospital-based cross-sectional study we investigated differences in the levels of serum atherosclerotic and fibrinolytic markers and the prevalence of metabolic syndrome (MS) among patients with four subtypes of cerebral infarctions. Blood samples were taken from 171 cerebral infarction inpatients to determine the levels of high-sensitivity C-reactive protein, serum total homocysteine, serum plasminogen activator inhibitor 1 and lipoprotein a. Subjects were also screened for MS. Atherothrombotic infarction was most prevalent, followed by lacunar and embolic infarction. The median length of hospital stay was longest for embolic infarcts. There were no statistically significant differences in serum marker concentrations. The proportion of MS varied significantly among the subtypes, and was highest among patients with embolic infarctions with the lowest high density cholesterol levels. MS was most prevalent among patients having undergone embolic events that are reported to have the worst prognoses. Further epidemiologic studies are needed to better understand the characteristics and differences in the etiology of cerebral infarction subtypes. PMID- 18411051 TI - Midpoint fixation task: quantitative assessment of visual neglect. AB - We present a method for assessing visual neglect using oculography, the "midpoint fixation" task. In the test, patients showed rightward deviation of the fixation point. Pure bias in visual neglect was considered to be assessed well by this method. Newly designed stimuli that lead to stable fixation and an effective calibration procedure are proposed. The method is suggested as useful in the assessment of visual neglect. PMID- 18411052 TI - Limbic encephalitis - a review. AB - The clinical features of limbic encephalitis are diverse and early diagnosis of the disorder is frequently difficult. Four patients with limbic encephalitis are described. An antineuronal antibody was identified in three of these patients. Antibodies directed against voltage-gated potassium channels, the N-methyl-D aspartate receptor and an unidentified neuropil antigen were each found in one patient. The fourth patient had multifocal paraneoplastic encephalitis associated with small cell lung cancer. The clinical and imaging findings associated with these antibodies and the other antineuronal antibodies described in patients with limbic encephalitis are reviewed. An approach to the diagnosis and management of limbic encephalitis is presented. PMID- 18411053 TI - Splenic angiosarcoma metastasis to the brain. AB - Angiosarcoma is a rare primary non-haematolymphoid malignant neoplasm of the spleen. This neoplastic process has a poor prognosis, with disease usually widespread by the time of presentation. In the literature there has been only six case reports of cerebral metastasis from splenic angiosarcomas. We report a 61 year-old woman who presented with a right occipital metastasis of a splenic angiosarcoma, 5 years after initial diagnosis, treated by stereotactic occipital craniotomy and resection of metastasis. PMID- 18411054 TI - Resorbability of rigid beta-tricalcium phosphate wedges in open-wedge high tibial osteotomy: a retrospective radiological study. AB - The open-wedge high tibial osteotomy (OWHTO) is a well accepted treatment modality for patients with osteoarthritis of the medial compartment associated with genu varum. To fill in the osteotomy gap 30% macroporosity rigid beta tricalcium phosphate (beta-TCP) is frequently used as a stable resorbable bone substitute. However, the resorbability of these beta-TCP wedges is not known. The aim of this study was to investigate this. Twenty-one OWHTO procedures in seventeen patients were performed with the use of 30% macroporosity rigid beta TCP wedges. The osteotomies were fixed using an angle-stable locking plate. Conventional AP and lateral radiographs were examined in order to assess the resorbability of the 30% macroporosity rigid beta-TCP wedges as a function of time. A radiological classification system consisting of five phases was used to monitor the resorption of the 30% macroporosity rigid beta-TCP wedges. The mean duration of follow-up was 62 months (+/-23 range of 28-99). In all 21 cases, remnants of the 30% macroporosity rigid beta-TCP wedges were still present at maximum follow-up. Although the boundaries between 30% macroporosity rigid beta TCP wedges and bone remained slightly visible, all osteotomies were completely consolidated and full osseointegration took place. In 16 out of 21 knees the fixation system was removed after a mean duration of 32 months (+/-19 range of 6 62). In six out of 21 knees a conversion to a knee arthroplasty was performed after a mean duration of 56 months (+/-18 range of 37-82). The OWHTO did not interfere with the placement of knee prostheses. Complete resorption of 30% macroporosity rigid beta-TCP wedges did not take place up to 8 years after operation. PMID- 18411055 TI - Regulation of AMPA receptor localization in lipid rafts. AB - Lipid rafts are special microdomains enriched in cholesterol, sphingolipids and certain proteins, and play important roles in a variety of cellular functions including signal transduction and protein trafficking. We report that in cultured cortical and hippocampal neurons the distribution of lipid rafts is development dependent. Lipid rafts in mature neurons exist on the entire cell-surface and display a high degree of mobility. AMPA receptors co-localize and associate with lipid rafts in the plasma membrane. The association of AMPARs with rafts is under regulation; through the NOS-NO pathway, NMDA receptor activity increases AMPAR localization in rafts. During membrane targeting, AMPARs insert into or at close proximity of the surface raft domains. Perturbation of lipid rafts dramatically suppresses AMPA receptor exocytosis, resulting in significant reduction in AMPAR cell-surface expression. PMID- 18411057 TI - XtalView, protein structure solution and protein graphics, a short history. AB - From a user's point-of-view we are in the Golden Age of protein crystallographic software. In the past few decades, solving protein structures has gone from a task requiring man-months of effort to a process requiring minutes on an ordinary laptop with no human intervention required. The birth of XtalView coincided with the mainstream use of synchrotron radiation, seleno-Met phasing and it continues to be used in this age of robotic crystallization, Fed-Ex data collection and fully automated structure solution "pipelines". This article is a retrospective history of protein crystallographic computing and a discussion of the current state of the art. PMID- 18411056 TI - Purification and reconstitution of the connexin43 carboxyl terminus attached to the 4th transmembrane domain in detergent micelles. AB - In recent years, reports have identified that many eukaryotic proteins contain disordered regions spanning greater than 30 consecutive residues in length. In particular, a number of these intrinsically disordered regions occur in the cytoplasmic segments of plasma membrane proteins. These intrinsically disordered regions play important roles in cell signaling events, as they are sites for protein-protein interactions and phosphorylation. Unfortunately, in many crystallographic studies of membrane proteins, these domains are removed because they hinder the crystallization process. Therefore, a purification procedure was developed to enable the biophysical and structural characterization of these intrinsically disordered regions while still associated with the lipid environment. The carboxyl terminal domain from the gap junction protein connexin43 attached to the 4th transmembrane domain (TM4-Cx43CT) was used as a model system (residues G178-I382). The purification was optimized for structural analysis by nuclear magnetic resonance (NMR) because this method is well suited for small membrane proteins and proteins that lack a well-structured three dimensional fold. The TM4-Cx43CT was purified to homogeneity with a yield of approximately 6 mg/L from C41(DE3) bacterial cells, reconstituted in the anionic detergent 1-palmitoyl-2-hydroxy-sn-glycero-3-[phospho-RAC-(1-glycerol)], and analyzed by circular dichroism and NMR to demonstrate that the TM4-Cx43CT was properly folded into a functional conformation by its ability to form alpha helical structure and associate with a known binding partner, the c-Src SH3 domain, respectively. PMID- 18411058 TI - Simulation, projection and empathy. AB - Simulationists have recently started to employ the term "empathy" when characterizing our most basic understanding of other minds. I agree that empathy is crucial, but I think it is being misconstrued by the simulationists. Using some ideas to be found in Scheler's classical discussion of empathy, I will argue for a different understanding of the notion. More specifically, I will argue that there are basic levels of interpersonal understanding-in particular the understanding of emotional expressions-that are not explicable in terms of simulation-plus-projection routines. PMID- 18411059 TI - I move, therefore I am: a new theoretical framework to investigate agency and ownership. AB - The neurocognitive structure of the acting self has recently been widely studied, yet is still perplexing and remains an often confounded issue in cognitive neuroscience, psychopathology and philosophy. We provide a new systematic account of two of its main features, the sense of agency and the sense of ownership, demonstrating that although both features appear as phenomenally uniform, they each in fact are complex crossmodal phenomena of largely heterogeneous functional and (self-)representational levels. These levels can be arranged within a gradually evolving, onto- and phylogenetically plausible framework which proceeds from basic non-conceptual sensorimotor processes to more complex conceptual and meta-representational processes of agency and ownership, respectively. In particular, three fundamental levels of agency and ownership processing have to be distinguished: The level of feeling, thinking and social interaction. This naturalistic account will not only allow to "ground the self in action", but also provide an empirically testable taxonomy for cognitive neuroscience and a new tool for disentangling agency and ownership disturbances in psychopathology (e.g. alien hand, anarchic hand, anosognosia for one's own hemiparesis). PMID- 18411060 TI - Measurement of GABAA receptor binding in vivo with [11C]flumazenil: a test-retest study in healthy subjects. AB - [(11)C]Flumazenil is widely used in positron emission tomography (PET) studies to measure GABA(A) receptors in vivo in humans. Although several different methods have been applied for the quantification of [(11)C]flumazenil binding, the reproducibility of these methods has not been previously examined. The reproducibility of a single bolus [(11)C]flumazenil measurements was studied by scanning eight healthy volunteers twice during the same day. Grey matter regions were analyzed using both regions-of-interest (ROI) and voxel-based analysis methods. Compartmental kinetic modelling using both arterial and reference region input function were applied to derive the total tissue distribution volume (V(T)) and the binding potential (BP) (BP(P) and BP(ND)) of [(11)C]flumazenil. To measure the reproducibility and reliability of each [(11)C]flumazenil binding parameter, absolute variability values (VAR) and intraclass correlation coefficients (ICC) were calculated. Tissue radioactivity concentration over time was best modelled with a 2-tissue compartmental model. V(T) showed with all methods good to excellent reproducibility and reliability with low VARs (mean of all brain regions) (5.57%-6.26%) and high ICCs (mean of all brain regions) (0.83 0.88) when using conventional ROI analysis. Also voxel-based analysis methods yielded excellent reproducibility (VAR 5.75% and ICC 0.81). In contrast, the BP estimates using pons as the reference tissue yielded higher VARs (8.08%-9.08%) and lower ICCs (0.35-0.80). In conclusion, the reproducibility of [(11)C]flumazenil measurements is considerably better with outcome measures based on arterial input function than those using pons as the reference tissue. The voxel-based analysis methods are proper alternative as the reliability is preserved and analysis automated. PMID- 18411062 TI - Phylogeny and classification of the Old World Emberizini (Aves, Passeriformes). AB - The phylogeny of the avian genus Emberiza and the monotypic genera Latoucheornis, Melophus and Miliaria (collectively the Old World Emberizini), as well as representatives for the New World Emberizini, the circumpolar genera Calcarius and Plectrophenax and the four other generally recognized tribes in the subfamily Emberizinae was estimated based on the mitochondrial cytochrome b gene and introns 6-7 of the nuclear ornithine decarboxylase (ODC) gene. Our results support monophyly of the Old World Emberizini, but do not corroborate a sister relationship to the New World Emberizini. Calcarius and Plectrophenax form a clade separated from the other Emberizini. This agrees with previous studies, and we recommend the use of the name Calcariini. Latoucheornis, Melophus and Miliaria are nested within Emberiza, and we therefore propose they be synonymized with Emberiza. Emberiza is divided into four main clades, whose relative positions are uncertain, although a sister relation between a clade with six African species and one comprising the rest of the species (30, all Palearctic) is most likely. Most clades agree with traditional, morphology-based, classifications. However, four sister relationships within Emberiza, three of which involve the previously recognized Latoucheornis, Melophus and Miliaria, are unpredicted, and reveal cases of strong morphological divergence. In contrast, the plumage similarity between adult male Emberiza (formerly Latoucheornis) siemsseni and the nominate subspecies of the New World Junco hyemalis is shown to be the result of parallel evolution. A further case of parallel plumage evolution, between African and Eurasian taxa, is pointed out. Two cases of discordance between the mitochondrial and nuclear data with respect to branch lengths and genetic divergences are considered to be the result of introgressive hybridization. PMID- 18411061 TI - Reliability of fMRI for studies of language in post-stroke aphasia subjects. AB - Quantifying change in brain activation patterns associated with post-stroke recovery and reorganization of language function over time requires accurate understanding of inter-scan and inter-subject variability. Here we report inter scan variability measures for fMRI activation patterns associated with verb generation (VG) and semantic decision/tone decision (SDTD) tasks in 4 healthy controls and 4 aphasic left middle cerebral artery (LMCA) stroke subjects. A series of 10 fMRI scans was completed on a 4T Varian scanner for each task for each subject, except for one stroke subject who completed 5 and 6 scans for SDTD and VG, thus yielding 35 and 36 total stroke subject scans for SDTD and VG, respectively. Group composite and intraclass correlation coefficient (ICC) maps were computed across all subjects and trials for each task. The patterns of reliable activation for the VG and SDTD tasks correspond well to those regions typically activated by these tasks in healthy and aphasic subjects. ICCs for activation were consistently high (R(0.05) approximately 0.8) for individual tasks among both control and aphasic subjects. These voxel-wise measures of reliability highlight regions of low inter-scan variability within language circuitry for control and post-recovery stroke subjects. ICCs computed from the combination of the SDTD/VG data were markedly reduced for both control and aphasic subjects as compared with the ICCs for the individual tasks. These quantitative measures of inter-scan variability support the proposed use of these fMRI paradigms for longitudinal mapping of neural reorganization of language processing following left hemispheric insult. PMID- 18411063 TI - Actin 5C, a promising nuclear gene for spider phylogenetics. PMID- 18411064 TI - Origin and diversification of the Greater Cape flora: ancient species repository, hot-bed of recent radiation, or both? AB - Like island-endemic taxa, whose origins are expected to postdate the appearance of the islands on which they occur, biome-endemic taxa should be younger than the biomes to which they are endemic. Accordingly, the ages of biome-endemic lineages may offer insights into biome history. In this study, we used the ages of multiple lineages to explore the origin and diversification of two southern African biomes whose remarkable floristic richness and endemism has identified them as global biodiversity hotspots (succulent karoo and fynbos). We used parsimony optimization to identify succulent karoo- and fynbos-endemic lineages across 17 groups of plants, for which dated phylogenies had been inferred using a relaxed Bayesian (BEAST) approach. All succulent karoo-endemic lineages were less than 17.5 My old, the majority being younger than 10 My. This is largely consistent with suggestions that this biome is the product of recent radiation, probably triggered by climatic deterioration since the late Miocene. In contrast, fynbos-endemic lineages showed a broader age distribution, with some lineages originating in the Oligocene, but most being more recent. Also, in groups having both succulent karoo- and fynbos-endemic lineages, there was a tendency for the latter to be older. These patterns reflect the greater antiquity of fynbos, but also indicate considerable recent speciation, probably through a combination of climatically-induced refugium fragmentation and adaptive radiation. PMID- 18411065 TI - Validation of a Persian-version of Knee injury and Osteoarthritis Outcome Score (KOOS) in Iranians with knee injuries. AB - OBJECTIVE: To adapt culturally and validate Persian-version of the Knee injury and Osteoarthritis Outcome Score (KOOS) in a sample of Iranians with knee injuries. METHODS: Cultural adaptation included providing of forward and backward translations, quality rating and pilot testing. A sample of 147 patients with anterior cruciate ligament (ACL), meniscus and combined (ACL and meniscus) injuries was asked to complete two questionnaires including the KOOS and Short Form 36 Health Survey (SF-36). The KOOS was readministered to 54 patients 6-8 days after the first visit. Test-retest reliability and internal consistency were assessed, using Intraclass Correlation Coefficient (ICC) and Cronbach's alpha, respectively. Dimensionality was assessed, using item-scale correlation after correction for overlap and construct validity, using a priori hypothesized correlations with the SF-36. RESULTS: All patients found the Persian-version of the KOOS to be clear and unambiguous in pilot testing. Minimum ICC level of 0.70 was exceeded by all subscales with the exception of Sport and Recreation (Sport/Rec) subscale. Minimum Cronbach's alpha level of 0.70 was exceeded by all subscales with the exception of Symptoms and Knee-related Quality of Life (QoL). Minimum Spearman correlation coefficient of 0.40 for each item-scale was exceeded by 34 items. All priori hypotheses were supported by the presence of higher correlations between similar constructs than between dissimilar constructs of the KOOS and SF-36. CONCLUSION: The Persian-version of the KOOS is a culturally adapted, reliable and valid outcome measure to be used in Iranian patients with knee injuries, with its psychometric properties in agreement with the original versions. PMID- 18411067 TI - The CLIP/CLAP-HSQC: pure absorptive spectra for the measurement of one-bond couplings. AB - Heteronuclear residual dipolar one-bond couplings of organic molecules at natural abundance are most easily measured using t2 coupled HSQC spectra. However, inevitably mismatched transfer delays result in phase distortions due to residual dispersive antiphase coherences in such experiments. In this article, slightly modified t2 coupled HSQC experiments with clean inphase (CLIP) multiplets are introduced which also reduce the intensities of undesired long-range cross peaks. With the corresponding antiphase (CLAP) experiment, situations where alpha and beta components overlap can be resolved for all multiplicities in an IPAP manner. A comparison of the experiments using hard pulses and shaped broadband excitation and inversion pulses on the heteronucleus is given and potential spectral artefacts are discussed in detail. PMID- 18411066 TI - Varenicline ameliorates ethanol-induced deficits in learning in C57BL/6 mice. AB - Ethanol is a frequently abused drug that impairs cognitive processes such as learning. Varenicline, an alpha4beta2 nicotinic receptor partial agonist and alpha7 nicotinic receptor full agonist prescribed for smoking cessation, has been shown to decrease ethanol consumption. The current study investigated whether varenicline could ameliorate ethanol-induced deficits in learning and whether varenicline alters blood alcohol concentration in C57BL/6 mice. Conditioning consisted of two auditory conditioned stimulus (CS; 30s, 85dB white noise)-foot shock unconditioned stimulus (US; 2s, 0.57mA) pairings. For all studies, saline or ethanol (1.0, 1.5, 2.0g/kg i.p.) was administered 15min before training, and saline or varenicline (0.05, 0.1, 0.2mg/kg i.p.) was administered 60min before either training or testing. For blood alcohol analysis, saline or varenicline (0.1mg/kg) was administered 60min before collection, and saline or ethanol (1.0, 1.5, 2.0g/kg) was administered 15min before collection. Varenicline dose dependently ameliorated ethanol-induced conditioning deficits for all three doses of ethanol when administered before training but not when administered 24h later, before testing. In addition, varenicline did not alter blood alcohol concentration. The smoking cessation aid varenicline may have therapeutic uses for treating ethanol-associated disruptions in cognitive processes. PMID- 18411068 TI - Two novel mutations in the insulin binding subunit of the insulin receptor gene without insulin binding impairment in a patient with Rabson-Mendenhall syndrome. AB - Homozygous or compound heterozygous mutations within the insulin binding domain of the human insulin receptor (INSR) are usually associated with severe impairment of insulin binding leading to Donohue syndrome ("Leprechaunism"), which is characterized by excessive hyperglycemia with hyperinsulinism, pre- and postnatal growth retardation, distinct dysmorphism and early death. Missense mutations in the beta subunits are commonly associated with a milder impairment of insulin binding and milder phenotype with prolonged survival and less dysmorphism, the so called Rabson-Mendenhall syndrome. We report on a 13-year-old girl with Donohue syndrome like dysmorphism, hyperinsulinism and prolonged survival due to two novel INSR missense mutations within the insulin binding domain. Unexpectedly, insulin binding assays and investigations of activation of central insulin signaling pathways in fibroblasts revealed no significant alterations. Instead, immunofluorescence studies showed abnormal perinuclear distribution of the INSR alpha and beta subunits. Our data indicate that the quality of insulin binding activity is correlated with survival, not with the dysmorphic phenotype, and it is not always a valid parameter for predicting INSR mutations as proposed. PMID- 18411069 TI - Newborn screening for glutaric aciduria type I in Victoria: treatment and outcome. AB - Between October 2001 and September 2007, a total number of 391,651 neonates were screened in Victoria using Tandem Mass Spectrometry and 6 newborns were diagnosed as having GA I, giving an incidence of 1:65,275 (CI: 1:29,988=1:177,861). Another patient was diagnosed through cascade screening of children born before the implementation of the expanded newborn screening program. Patients were treated by mild protein restriction (2-2.5 g/kg/day) and carnitine supplementation when well, focussing on the aggressive management of intercurrent illnesses (temporary cessation of protein intake, increase in calorie intake, IV carnitine, aggressive anti febrile and anti infectious treatment), including prophylactic admissions to hospital. Overall, our patients had 35 admissions to hospital, of which 15 were in the first year of life. None had a post infectious dystonic syndrome. Neuropsychological examinations revealed normal to high cognitive and gross motor function in all patients but one, with some deficiencies in fine motor activities and different levels of speech abnormalities in all patients. Since therapeutic approaches for GA I, although not uniform, are well established and have been documented to be effective, newborn screening for this disorder should prove justified. A therapeutic approach of dietary modification, IV carnitine and aggressive treatment of intercurrent illness seems to prevent the severe neurological complications of GA I. More in-depth consideration of speech and language function is necessary to document specific deficits in children with GA I and plan proactive interventions. PMID- 18411070 TI - Experimental validation of the filtering approach for dose monitoring in proton therapy at low energy. AB - The higher physical selectivity of proton therapy demands higher accuracy in monitoring of the delivered dose, especially when the target volume is located next to critical organs and a fractionated therapy is applied. A method to verify a treatment plan and to ensure the high quality of the hadrontherapy is to use Positron Emission Tomography (PET), which takes advantage of the nuclear reactions between protons and nuclei in the tissue during irradiation producing beta(+)-emitting isotopes. Unfortunately, the PET image is not directly proportional to the delivered radiation dose distribution; this is the reason why, at the present time, the verification of depth dose profiles with PET techniques is limited to a comparison between the measured activity and the one predicted for the planned treatment by a Monte Carlo model. In this paper we test the feasibility of a different scheme, which permits to reconstruct the expected PET signal from the planned radiation dose distribution along beam direction in a simpler and more direct way. The considered filter model, based on the description of the PET image as a convolution of the dose distribution with a filter function, has already demonstrated its potential applicability to beam energies above 70 MeV. Our experimental investigation provides support to the possibility of extending the same approach to the lower energy range ([40, 70] MeV), in the perspective of its clinical application in eye proton therapy. PMID- 18411071 TI - Distractor P3 is associated with attentional capture by stimulus deviance. AB - OBJECTIVE: A simple distractor elicits a large P3 when the standard and target are difficult to discriminate in the three-stimulus oddball paradigm. This study investigated whether the distractor P3 reflects attentional capture by stimulus deviance or cognitive interference with maintaining the standard representation. METHODS: Event-related brain potentials were recorded from 12 participants who performed a visual three-stimulus oddball paradigm. Four task conditions were defined by a combination of two presentation types of distractor stimuli (central or bilateral) and two levels of standard/target discrimination difficulty (easy or difficult). Bilateral distractors had stimulus deviance but did not interfere with maintenance of the standard representation. RESULTS: Central distractors elicited a P3, the amplitude of which was larger in the difficult task than in the easy task. In contrast, bilateral distractors elicited a large P3 in both the easy and difficult tasks. CONCLUSIONS: Distractor P3 reflects attentional capture by stimulus deviance, rather than cognitive interference with maintaining the standard representation. SIGNIFICANCE: This is the first report showing that simple distractors can elicit large anteriorly distributed P3 in an easy task. The present findings contribute to the clinical application of distractor P3 to assess the cognitive function of deviant processing. PMID- 18411072 TI - Rhythmic arm cycling suppresses hyperactive soleus H-reflex amplitude after stroke. AB - OBJECTIVE: Rhythmic arm cycling movement suppresses the amplitude of soleus H reflexes in neurologically intact participants. This suppression is greater when the movement frequency is increased. If rhythmic arm movement can still suppress the amplitude of H-reflexes in the legs after stroke, it could potentially be used as a rehabilitation technique to reduce exaggerated reflexes such as those occurring in spasticity. The purpose of this study was to test for maintenance of this suppressive effect after stroke. Since a portion of the effect of arm cycling had previously been ascribed to subcortical and spinal mechanisms, we hypothesized that the suppressive effect of arm cycling would be partially maintained after stroke. METHODS: Participants with history of single chronic (> 6 months) stroke performed rhythmic arm cycling at approximately 1 Hz and also at the highest frequency possible ( approximately 1.5 Hz). Soleus H-reflexes were evoked in the more and less affected legs simultaneously and full recruitment curves obtained. RESULTS: H-reflex amplitudes in both the more and less affected legs of stroke participants were significantly suppressed during arm cycling. However, the extent of the suppression is weaker compared to neurologically intact and age-matched subjects. CONCLUSIONS: Neural activity related to arm cycling can still access interlimb pathways after stroke and activate spinal control mechanisms leading to suppression of H-reflex amplitudes. SIGNIFICANCE: The suppressive effect of arm cycling could be exploited in the modification of exaggerated muscle afferent reflexes in leg muscles after stroke. Whether this has a significant effect on modulation of spasticity requires further substantiation. PMID- 18411073 TI - Expression of the chick Sizzled gene in progenitors of the cardiac outflow tract. AB - The secreted Frizzled receptor related proteins (Sfrp's) belong to a protein family that comprises antagonists and modifiers of Wnt and BMP signalling. Here we report the isolation and expression pattern of the Sfrp gene "Sizzled" in the chick. Sizzled genes, as well as the closely related crescent genes, exist in the genomes of fishes, frogs and chicks, but not of mammals. The chicken Sizzled gene (Szl) is initially expressed in the anterior endoderm of gastrulating and early head fold embryos. An additional, separate expression domain develops at the posterior end of the embryo from the head process stage onwards. Szl transcripts are then detected in precardial mesodermal cells, are transiently transcribed in the straight heart tube, and later prominently in the splanchnic mesoderm surrounding the arterial pole of the developing heart, the anterior heart field. These cells are subsequently recruited to form the cardiac outflow tract. cSzl expression is downregulated when the septation of the outflow tract by neural crest derived cells begins. PMID- 18411074 TI - Neonatal anemia. AB - Neonatal anemia and the need for red blood cell (RBC) transfusions are very common in neonatal intensive care units. Neonatal anemia can be due to blood loss, decreased RBC production, or increased destruction of erythrocytes. Physiologic anemia of the newborn and anemia of prematurity are the two most common causes of anemia in neonates. Phlebotomy losses result in much of the anemia seen in extremely low birthweight infants (ELBW). Accepting a lower threshold level for transfusion in ELBW infants can prevent these infants being exposed to multiple donors. PMID- 18411075 TI - Treatment of muscular contraction deformities with botulinum toxin type A after latissimus dorsi flap and sub-pectoral implant breast reconstruction. AB - Unusual and probably underestimated complications following breast reconstruction with a latissimus dorsi (LD) flap and sub-pectoral implant are the LD muscle twitching and the breast contour deformities from pectoralis major (PM) muscle contraction. Surgical muscle denervation is usually indicated for these complications. Botulinum toxin A (BTX-A) infiltration has been described in reducing breast contour deformity in sub-pectoral implant after breast augmentation or reconstruction. Between January 2002 and April 2006, 71 consecutive patients underwent delayed unilateral breast reconstructions with LD flap and sub-pectoral implant after mastectomy. All patients reporting discomforting signs and symptoms from muscle contraction in the reconstructed breast were included in this prospective study. Thirteen patients (18.3%) were selected and treated with BTX-A percutaneous local injections. Signs and symptoms were evaluated, after 4, 8 and 12 months, by the patients and by a panel of three physicians not involved in the study, using a five-point scale. During the study period all patients reported a decrease or disappearance of the signs and symptoms. After 12 months, 11 patients received three BTX-A infiltrations, demonstrating considerable improvements compared to the pre-treatment status. Wilcoxon matched pairs rank sum test showed a statistical difference between pre treatment and post-treatment scores after 14 days (P<0.01) and 12 months (P<0.001). Our experience shows that muscular contraction deformities after breast reconstruction with a LD flap plus implant are not uncommon complications. The use of BTX-A infiltrations is an effective, not surgical, low cost and low risk procedure to treat these complications. It is an easy procedure to be performed on an outpatient basis with a temporary effect but safely repeatable and reproducible; it avoids hospitalisation or further surgical procedures and demonstrates tolerable latency with satisfactory outcomes. PMID- 18411076 TI - The population of health economic models is critical. PMID- 18411077 TI - Hemopoietic growth factors show promise as therapeutic agents. AB - Hemopoietic growth factors such as erythropoietin and colony-stimulating factors may have important therapeutic applications. PMID- 18411078 TI - Primary hyperparathyroidism. AB - Many studies of asymptomatic primary hyperparathyroidism by noninvasive bone densitometry are beginning to show that demineralization is relatively common. This patient showed a pattern that is most typical of primary hyperparathyroidism, namely, normal bone mineral density of the vertebral spine and significant reduction of bone mineral density of the distal radius. PMID- 18411079 TI - Familial benign hypercalcemia. AB - Familial benign hypercalcemia closely mimics asymptomatic primary hyperparathyroidism. It needs to be distinguished from hyperparathyroidism, as surgical treatment is not indicated outside of exceptional circumstances. PMID- 18411080 TI - Bone-derived proteins. AB - The diagnosis and treatment of metabolic bone disease has been limited by a lack of understanding of bone cell physiology and of suitable markers for various bone cell functions in different diseases. The identification of noncollagenous proteins and their measurement in serum has added to our knowledge. At this stage, of all the bone-derived proteins, bone Gla protein (BGP) is apparently the most useful, but it is best used in conjunction with other tests. The diagnostic usefulness of other noncollagenous proteins, procollagen, and local factors still remains to be documented. PMID- 18411081 TI - Calcium signaling and gonadotropin secretion. AB - Agonist activation of pituitary gonadotrophs by gonadotropin-releasing hormone (GnRH) stimulates rapid InsP(3)-dependent peaks of calcium mobilization and luteinizing hormone (LH) release, followed by sustained increases in calcium influx and hormone secretion. Receptor-mediated calcium entry through L-type and dihydropyridine-insensitive calcium channels accounts for the sustained elevation of cytosolic calcium during GnRH action, and for most of the gonadotropin secretory response. Protein kinase C contributes to the phase of sustained LH release from GnRH-stimulated gonadotrophs, and also to gonadotropin synthesis. Calcium-dependent inactivation of L channels occurs during GnRH action, and appears to be a primary factor in the onset of desensitization of gonadotropin secretion. PMID- 18411082 TI - Regulation of pulmonary surfactant protein synthesis in fetal lung: a major role of glucocorticoids and cyclic AMP. AB - Augmented synthesis of the lipoprotein, pulmonary surfactant, is initiated in fetal lung toward the end of-gestation. Inadequate surfactant synthesis by the lungs of premature infants can result in respiratory distress syndrome, the leading cause of neonatal morbidity and mortality in developed countries. The surfactant-associated proteins act with surfactant glycerophospholipids to reduce alveolar surface tension, and mediate the reutilization of secreted surfactant components by type II cells. Genes encoding the surfactant proteins SP-A, SP-B, and SP-C have been isolated and characterized. Recent findings suggest that surfactant protein gene expression in fetal lung is under multifactortal control and is regulated by glucocorticoids, cAMP, growth factors, and insulin. PMID- 18411083 TI - Signal transduction and the ras gene family: molecular switches of unknown function. AB - The ras family consists of 20 or more genes that encode small GTP/GDP-binding proteins, of 20-26 kDa, the functions of which are unknown. This article discusses possible roles of the ras proteins in signal transduction and the interaction of p21(ras) and other members of the ras family with GTPase accelerating proteins (GAPS) that may be regulatory elements of the signaling machinery. PMID- 18411084 TI - Mammosomatotropes: current status and possible functions. AB - Recent studies show that mammosomatotropes-cells that secrete both growth hormone and prolactin-are common in normal pituitary tissue. It is proposed that these dual hormone secretors serve as transitional cells for the differentiation of classical mammotropes during development, and for the functional interconversion of growth hormone and prolactin secretors in adults. PMID- 18411085 TI - Thyroid and steroid hormone regulation of proto-oncogene expression. AB - In contrast to the established effects of peptide growth factors on specific proto-oncogene expression, the actions of steroid and thyroid hormones are less clearly defined. However, there is increasing evidence that these hormones, acting through structurally related DNA-binding nuclear receptor proteins, influence proto-oncogene expression. This influence may determine the function of steroid and thyroid hormones in regulation of cell proliferation and maturation, and provide insight into the role of these hormones in oncogenesis. PMID- 18411086 TI - Parathyroid hormone-like peptide: molecular characterization and biological properties. AB - Parathyroid hormone-like peptide (PLP) is a newly discovered novel peptide that interacts with renal and osseous parathyroid hormone (PTH) receptors through its amino-terminal sequence, which is homologous with PTH. Human PLP is the product of a complex single-copy gene, existing in several isoforms which result from alternative exon usage. The peptide was implicated as a causal agent in the hypercalcemia associated with a variety of malignancies, but it may also modulate calcium homeostasis in some normal physiological states and mediate cellular growth or differentiation. PMID- 18411087 TI - The immunogenetics of insulin-dependent (Type I) diabetes. AB - Insulin-dependent diabetes develops when a genetically predisposed individual is exposed to an as-yet-unknown environmental insult. A major part of genetic susceptibility to the disorder is encoded close to or within the HLA-DQ region, but non-HLA-linked genes are also implicated. PMID- 18411088 TI - Experimental uses of neurohypophysial hormone analogs. AB - Structural analogs of neurohypophysial peptides with specific agonistic and antagonistic activities are valuable tools for research on these hormones and their receptors. They may also provide new or improved therapeutic agents. PMID- 18411089 TI - Endocrine alterations associated with the abuse of cocaine. AB - Cocaine-induced disturbances in central nervous system neurotransmitter function may also occur in tuberoinfundibular neurons and these alterations may result in secondary clinical endocrine abnormalities. PMID- 18411090 TI - Is insulin a factor in the genesis of the vascular complications of diabetes? AB - Several recent studies report that insulin may be a pathogenic factor in cardiovascular disease and the vascular complications of diabetes. The route of administration of insulin may be partially responsible for vascular side effects that occur in the management of diabetes. These side effects may be overcome by the development of a more physiologic route of administration than by subcutaneous injection. PMID- 18411091 TI - Polycystic ovary syndrome. AB - This case illustrates the very common endocrine problem of polycystic ovary syndrome (PCOS). The diagnosis was not clear initially because of the absence of clinical evidence of androgen excess and of a normal random serum luteinizing hormone (LH) concentration. Futher investigations, however, confirmed polycystic ovaries on ultrasound and revealed a raised serum LH and elevated testosterone despite the lack of hirsutism. The patient's anovulatory infertility was treated by low dose human menopausal gonadotrophin (HMG), which resulted in ovulation of a single dominant follicle and pregnancy in the first cycle of treatment. This article discusses the endocrine features of hirsute and nonhirsute patients with PCOS, the choice of treatment for induction of ovulation, and comments on the possible mechanisms underlying PCOS. PMID- 18411092 TI - Cyclic GMP and the second messenger hypothesis. AB - A plasma membrane form of guanylate cyclase appears to contain a single transmembrane domain that divides the protein into a highly conserved intracellular domain and a variable extracellular domain. Various extracellular peptides can bind directly to guanylate cyclase to increase the production of the second messenger, cyclic GMP. PMID- 18411094 TI - Receptor-effector coupling by G-proteins implications for endocrinology. AB - Discovered serendipitously in the course of studies on the mechanism of glucagon stimulation of hepatic cyclic AMP formation, G-proteins have emerged as an expanding family of signal transducers, coupling diverse receptors and effectors. Quantitative and/or qualitative changes in G-proteins may profoundly affect hormone action, and can lead to clinically apparent endocrine dysfunction. PMID- 18411093 TI - Molecular genetics of congenital adrenal hyperplasia. AB - More than 95% of cases of congenital adrenal hyperplasia are attributable to steroid 21-hydroxylase (21-OH) deficiency. In normal individuals, there are usually two 21-OH genes on each chromosome 6, a functional 21-OH gene-CYP21B-and a closely related 21-OH pseudogene-CYP21A-which is defective in expression. Recent advances have shown that the pathologic mutations that contribute to 21-OH deficiency arise as a consequence of unequal crossover and gene conversion-like mechanisms that involve sequence interaction between the normally functional 21 OH gene and its pseudogene. PMID- 18411095 TI - Familial multiple endocrine neoplasia type 1 Mutation of a tumor suppressor gene. AB - Familial multiple endocrine neoplasia type 1 (FMENI) is caused by mutation of a gene on the long arm of chromosome 11. Inactivation of both alleles at this locus in one cell is thought to cause loss of growth inhibition and development of a monoclonal tumor. PMID- 18411096 TI - Distinct functional domains on the insulin receptor beta-subunit Do they provide a molecular basis for "selective" insulin resistance? AB - The insulin receptor is seen as having a number of structurally and functionally distinct domains. Modifications of particular domains may lead to the partial crippling of receptor function, which could give rise to selective insulin resistant states. PMID- 18411097 TI - The plasma membrane and thyroid hormone entry into cells. AB - A growing body of evidence indicates that many cells accumulate thyroid hormone by energy-dependent receptor-mediated pathways. Their contributions to the regulation of thyroid hormone metabolism and action are now being addressed. PMID- 18411098 TI - Growth factors and pituitary tumors. AB - Growth within the anterior pituitary gland is probably controlled by several interacting extracellular messenger molecules, including hypothalamic peptides, target gland hormones, and several growth factors acting in autocrine or paracrine fashion. Adenoma formation may result from abnormal production of such factors or their specific cellular receptors, loss of local inhibitory influences, activation of the intracellular secondary message pathways conveying the mitogenic signal to the nucleus, or deregulation of the nuclear processes controlling mitosis. PMID- 18411099 TI - Complexity of steroid hydroxylase gent expression in the adrenal cortex A microcosm of regulated transcription. AB - Regulation of the expression of steroid hydroxylase genes in the adrenal cortex involves mechanisms required for maintenance of optimal steroidogenesis, tissue specificity, and ontogeny of the steroidogenic pathway. Evaluation of the molecular basis of this complexity promises to unfold new aspects of regulated eukaryotic gene expression. PMID- 18411100 TI - Pituitary cells producing more than one hormone human pituitary adenomas. AB - The existence of cells capable of producing more than one hormone in nontumorous human adenohypophyses and pituitary adenomas has been conclusively proved. In light of the evidence, current concepts on pituitary structure, function, and regulation as well as adenoma cytogenesis and classification have to be reconsidered. PMID- 18411101 TI - 44-2C cells a neuroendocrine model to study neuropeptide, oncogene, and growth factor gene regulation. AB - The neuroendocrine 44-2C cells synthesize, secrete, and manifest differential regulation of calcitonin (CT), CT gene-related peptide (CGRP), neurotensin (NT), and somatostatin (SS). These cells maintain differentiated function when grown in serum-free, growth factor and hormone-deprived milieu. The cells continue to secrete CT, CGRP, NT, and SS and differentially respond to cellular secretagogues. In serum-free cultures, the cells produce biologically active acidic fibroblast growth factor (aFGF) and differentially regulate the expression of the protooncogene fos (c-fos). PMID- 18411102 TI - The role and regulation of testicular opioids. AB - Endogenous opioids are present in the male gonad and may be involved in the local control of testicular function. Testicular opioids are mainly produced in Leydig cells and exert direct paracrine inhibitory actions on the tubule, which in turn may have indirect Sertoli-cell-mediated effects on Leydig cell steroidogenesis. Gonadotropins and CRF stimulate production of gonadal opioids, and the presence of a gonadotropin milieu appears to be a requisite for the inhibitory paracrine actions of opioids on steroidogenesis. The recognition of such multiple intratesticular hormonal interactions should provide insights into the mechanisms involved in idiopathic infertility in men. PMID- 18411103 TI - Hippocampal-steroid complexity. AB - Interaction of glucocorticoid hormones with hippocampal receptors can modulate complex endocrine and behavioral processes. A current article on hippocampal neuronal degeneration in response to long-term adrenalectomy emphasizes the complex and specific cellular mechanisms that underlie such effects. PMID- 18411104 TI - Congenital adrenal hyperplasia. AB - An autosomal recessive disorder, congenital adrenal hyperplasia, results from a deficiency in the activity of one of the five enzymes required for cortisol biosynthesis. More than 90% of cases are related to deficiency in 21-hydroxylase enzyme activity, which is required for the conversion of 17OH-progesterone to 11 deoxycortisol. Treatment of congenital adrenal hyperplasia consists of steroid replacement to ensure normal growth and reproductive potential. PMID- 18411105 TI - Useful clinical assays for vitamin D metabolites. AB - Advances in our understanding of the physiology of vitamin D and its importance in health and disease have depended on the accurate measurement of its metabolites in blood. Assays that were once cumbersome and insensitive are now performed easily, are highly sensitive, reproducible, and relatively inexpensive. The availability of these modern techniques has facilitated the clinical diagnosis and treatment of patients. PMID- 18411106 TI - Mutations of the human insulin receptor gene. AB - Mutations in the insulin receptor gene have been identified in patients with genetic forms of insulin resistance. These mutations provide insight into structure-function relationships of the insulin receptor, and also into the causes of insulin resistance in human disease. PMID- 18411107 TI - Clinical applications of somatostatin analogs. AB - The somatostatin analog Sandostatin is successfully used in the treatment of metastatic endocrine pancreatic tumors, carcinoids, and acromegaly. In addition, somatostatin receptors are also present on other tumors in man, therefore making it possible to demonstrate these tumors by the administration of (123)I-coupled to a somatostatin analog. PMID- 18411108 TI - Target tissue specificity of mineralocorticoids. AB - Mineralocorticoid receptors have equivalent, high affinity for aldosterone and for the physiological glucocorticoids cortisol and corticosterone, which circulate at much higher concentrations. In physiologic mineralocorticoid target tissues, glucocorticoids are excluded from mineralocorticoid receptors by conversion to their receptor-inactive 11-keto metabolites, by the enzyme 11beta hydroxysteroid dehydrogenase. Aldosterone, in contrast, escapes metabolism by cyclizing its 11-OH group with the reactive -CHO at position 18. In most endocrine systems, specificity is conferred in large part by the ability of the receptor to distinguish between signal and noise. Why, for mineralocorticoid action, specificity is prereceptor-a nonmetabolized signal, and an enzyme that excludes noise-remains to be explored. PMID- 18411109 TI - The role of fibroblast growth factors in the central nervous system. AB - The fibroblast growth factors are well-characterized mitogens that are found in the central nervous system (CNS). Their physiological roles are not yet known, but increasing evidence suggests their involvement in CNS development, injury responses, and possibly oncogenesis. PMID- 18411110 TI - Characterization of a cyclic AMP regulatory element DNA-binding protein. AB - Signal transduction pathways converge ultimately at the level of transcriptional activation to produce specific patterns of gene expression in response to environmental stimuli. The initiation of transcription mediated by these signaling pathways is regulated by the coordinate expression and/or activation of specific transcription factors that bind to the control regions of eukaryotic genes. Specific insights into the mechanisms underlying transcriptional activation have arisen from the studies of the structure and functions of eukaryotic transcription factors. One of these factors, a cyclic (c)AMP response element binding protein (CREB), has only recently been discovered and appears to play a key role in the regulation of gene expression in response to the activation of the cAMP-dependent protein kinase A. The transcription factors related to CREB, c-jun and c-fos, mediate transcriptional responses to activators of protein kinase C. PMID- 18411111 TI - Second messengers of insulin action. AB - The molecular events involved in coupling the insulin receptor to the regulation of cellular metabolism remain unknown. Recent studies indicate that some of insulin's actions may be mediated by a novel oligosaccharide. This molecule is generated in cells by the insulin-dependent hydrolysis of a novel membrane glycolipid, termed a glycosyl-phosphatidylinositol. This glycolipid is structurally similar to a newly described protein anchor. The evaluation of the hormonal regulation of this new glycolipid may yield information on a new mechanism of signal transduction. PMID- 18411112 TI - Parathyroid hormone-receptor interactions. AB - Identification of sites within the antagonist peptide of parathyroid hormone (PTH) that are "tolerant" of a wide range of amino acid substitutions has led to the design of new PTH antagonists. These antagonists have increased potency because of substitution, at appropriate sites, of amino acids that increase the interaction of the ligand with its receptor but do not cause signal transduction. Similar modifications in the parathyroid hormone-related protein (PTHrP) antagonist led to antagonists with increased potency. Further, the partial agonism of this analog could be removed by exchange of residues between PTH and PTHrP. PMID- 18411113 TI - Peptide YY and neuropeptide Y in the gut: availability, biological actions, and receptors. AB - Intensive research on the actions of peptide YY(PYY) and neuropeptide Y (NPY) on the gut has been stimulated by findings of potent anti-secretory effects in small intestine and the discovery of their common receptor in this tissue. There is evidence that the hormone PYY and the neurotransmitter NPY are involved in the regulation of fluid and electrolyte secretion, motility, and blood flow in the intestine. Intestinal PYY/NPY receptors may have pharmacological value for the treatment of diarrhea. PMID- 18411114 TI - Control of thyroid cell and follicle growth: recent advances and current controversies. AB - The regulation of cell proliferation within the thyroid follicle is a coordinated and finely balanced process involving integration of the action of pituitary thyrotropin with the effects of permissive and inhibitory growth factors of autocrine and paracrine origin. Our understanding of the cellular interactions and intracellular signalling processes involved in thyroid follicular growth control has been considerably assisted by in vitro cell culture techniques that enable thyroid follicular cells to be maintained and studied under conditions closely approximating those in vivo. PMID- 18411115 TI - Cloning of the LH/CG receptor: implications for a unique G-protein coupled receptor. AB - The proteins encoded by the rat and porcine luteinizing hormone (LH)/chorionic gonadotropin (CG) receptor cDNAs appear to be unique members of the G -protein coupled receptor family. Although they have the characteristic seven transmembrane domains, the LH/CG receptors have relatively low homology to other members o f this family, do not have a G-protein-coupling domain corresponding to that of the a-adrenergic receptor, and contain an unusual long extracellular domain. Experience in the molecular dissection of this receptor family will help guide investigation of the LH/CG receptor's functional domains. Further studies are needed to clarify the origin and significance of the various mRNA species hybridizing on Northern blot analyses. Finally, the rat and porcine receptor cDNAs should permit cloning of the human LH/CG receptor, as well as cloning of the thyroid-stimulating hormone and follicle-stimulating hormone receptors. PMID- 18411116 TI - Hirsutism. AB - The clinical presentation and management of the hirsute woman is illustrated in cases of idiopathic hirsutism, polycystic ovarian disease, and nonclassical congenital adrenal hyperplasia. Therapeutic regimens manage hyperandrogenic signs, induce ovulation, and protect the endometrium. PMID- 18411117 TI - The brain angiotensin system: insights from mapping its components. AB - Mapping of components of the angiotensin (Ang) system in the brain suggests that it serves multiple central roles, including regulation of fluid and electrolyte balance, central autonomic control, and pituitary hormone release. PMID- 18411118 TI - "APUD" cells: fact and fiction. AB - Pearse's "APUD" cell theory helps define the ontogeny and differentiation programs of small polypeptide hormone-secreting cells of diverse tissue origins. Cells with such features may arise through one of multiple related pathways of epithelial cell differentiation ongoing in tissues of all embryonic lineages. PMID- 18411119 TI - Antibody markers for type 1 diabetes: the issues. AB - The detection of islet-cell-specific autoantibodies before clinical onset may mean that diabetes mellitus can be predicted, with important implications for targeting those to whom preventive treatment might be given. A large number of such markers have been proposed, but only high-titer islet cell antibodies have so far demonstrated the specificity and sensitivity to be of potential clinical value. PMID- 18411120 TI - The role of sulfation in thyroid hormone metabolism. AB - Sulfate conjugation is a significant metabolic reaction for thyroxine and especially so for triiodothyronine and lower iodothyronines in rats. Triiodothyronine sulfation has also been demonstrated in humans. Sulfation accelerates the deiodinative breakdown of iodothyronines by the type I iodothyronine deiodinase in liver and thus represents a rate-limiting step in one of the elimination pathways of thyroid hormone. PMID- 18411121 TI - Regulatory elements of the pro-opiomelanocortin gene: pituitary specificity and glucocorticoid repression. AB - A short 543-bp fragment o f the pro-opiomelanocortin gene is sufficient for pituitary-specific expression and, in the anterior pituitary gland, for repression of pro-opiomelanocortin transcription by glucocorticoids. Within this 5'-flanking fragment of the gene, multiple regulatory elements contribute to tissue-specific expression and a single glucocorticoid receptor binding site acts as a "negative glucocorticoid response element." The current model for glucocorticoid repression depends on the mutually exclusive binding of the glucocorticoid receptor and of a positive transcription factor, the chicken ovalbumin upstream promoter element transcription factor, to overlapping DNA sequences within the negative glucocorticoid response element. PMID- 18411122 TI - Specificity of the mineralocorticoid receptor: crucial role of 11beta hydroxysteroid dehydrogenase. AB - Research on the enzyme 11beta-hydroxysteroid dehydrogenase, initially performed on congenital deficiency of the enzyme, and later work on deficiency of the enzyme after licorice and carbenoxolone administration, led to the hypothesis that 11beta-hydroxysteroid dehydrogenase conferred specificity on the mineralocorticoid receptor. PMID- 18411123 TI - Advances in Paget's disease treatment. PMID- 18411124 TI - Interleukin-6 a putative mediator of anterior pituitary hormone secretion during the stress of infection? AB - Interleukin-6 may have an important role as a cytokine involved in the neuroendocrine-immune response. It is produced by adenohypophysis and may modulate pituitary hormone release during the stress of infection. PMID- 18411125 TI - Management of constitutional delay in growt and puberty. AB - Constitutional delay in growth and puberty is a variant of normal growth and development that can cause a significant degree of psychological disturbance in otherwise healthy children, and is most often seen in boys of pubertal age. Careful assessment is necessary to rule out other endocrine or nonendocrine diseases. In some patients, therapy with oxandrolone or testosterone may be necessary to advance growth and/or pubertal development and thereby prevent serious psychological disturbance that can persist even into adult life. In the majority, however, reassurance will usually suffice. PMID- 18411126 TI - Advances in techniques for measurement of parathyroid hormone Current applications in clinical medicine and directions for future research. AB - Immunometric assays that measure intact parathyroid hormone are replacing traditional radio immunoassays because they provide better discrimination of parathyroid gland function in classic disorders of calcium homeostasis, although radio immunoassays and bioassays continue to have roles in some clinical and research situations. The sensitivity and high precision of Immunometric assays will enable definition of parathyroid hormone's role in chronic and subtle disturbances of calcium and bone homeostasis, such as bone loss and calcium stone disease, in addition to assisting the clinician in the differential diagnosis of hypercalcemic and hypocalcemic disorders. PMID- 18411127 TI - Oncogenes, antioncogenes, and the regulation of cell growth. AB - A variety of proto-oncogenes are present in normal cells, and many of these genes are expressed in different cell types in a tissue-specific and developmentally specific fashion. Although proto-oncogenes have normal functions, apparently related to cell proliferation and differentiation, these genes are able to cause cancer when they are expressed inappropriately (usually as a result of a mutation). Most oncogene mutations promote tumor growth by inducing autonomous activity of proteins, which normally transmit growth signals that are triggered by extracellular factors. Studies of oncogenes and suppressor genes (antioncogenes) have greatly contributed to our understanding of the regulation of normal cell growth by focusing on the molecular mechanisms by which the signals for cell proliferation exert their effects. PMID- 18411128 TI - Therapeutic potential of insulinlike growth factor i. AB - Human recombinant insulinlike growth factor I is a promising therapeutic agent for diseases characterized by relative insulin resistance, e.g., diabetes mellitus, obesity, and hypertriglyceridemia, since it suppresses growth hormone, insulin, C-peptide, and triglyceride levels and lowers the total cholesterol to high-density lipoprotein-cholesterol ratio. Moreover, insulinlike growth factor administration increases kidney function in healthy subjects (glomerular filtration rate, renal plasma flow) and may prove useful in the treatment of degenerative disorders of cartilage and bone (arthrosis, osteoporosis) as well as in catabolic states. PMID- 18411129 TI - Proinsulin trafckin and processing in the pancreatic B cell. AB - Insulin is synthesized as a precursor, preproinsulin, in the rough endoplasmic reticulum of the pancreatic B cell. The combination of precursor processing and the movement of products in vesicles from one subcellular compartment to the next results in insulin becoming stored in secretory granules ready for release in response to a secretagogue. PMID- 18411130 TI - Human autoimmune thyroid disease A mechanistic update. AB - Autoimmune thyroid disease refers to a group of common, often familial, disorders of thyroid function, which are associated with disturbances in both cellular and humoral immunity. Current research into autoimmune thyroid disease is characterizing specific target antigens and the cellular and molecular events that lead to tissue-specific immune dysfunction. The thyroid gland is an accessible and useful model for the study of human autoimmune disease. This review highlights some of the more active research areas. PMID- 18411131 TI - The use of immunoradiometric assay for the measurement of ACTH in human plasma. AB - Bioassay and immunoassay techniques for the measurement of ACTH in human plasma have provided sensitive and specific results but have also met with some skepticism as to their reliability in some clinical circumstances. The recent development of a supersensitive two-site immunoradiometric assay for ACTH may resolve sole of the limitations of previous assays and greatly facilitate the evaluation of pituitary-adrenal disorders. PMID- 18411132 TI - Clinically useful calcitonin assays. AB - Calcitonin assays are essential tools for research into calcitonin (CT) and C cell physiology and pathophysiology. Several existing radio immunoassays for CT, based on polyclonal antisera, are generally reliable for detection and follow-up of patients having medullary thyroid carcinoma. Occasionally, however, these assays suffer from problems of specificity and sensitivity. Two-site immunometric techniques for CT offer potential for great improvement in sensitivity, specificity, and turnaround time over those based on classical competitive binding radioimmunoassays. PMID- 18411133 TI - Cushing's syndrome: difficulties in diagnosis. AB - Establishment of the precise cause in a patient with Cushing's syndrome remains a major clinical challenge. The following case of a female patient with cyclical Cushing's syndrome illustrates how recent advances such as the introduction of the corticotropin-releasing hormone (CRH) test (in determining the cause of ACTH production) and high-resolution computed tomographic scanning may help in the diagnosis of Cushing's syndrome. As this case study illustrates, precise diagnosis requires thorough initial investigation and possible successive testing. PMID- 18411134 TI - Pregnancy and autoimmune thyroid disease. AB - Immunological and endocrine changes related to pregnancy influence the course and development of autoimmune thyroid disease (AITD). Conversely, the presence of AITD can affect both the mother and her child-both in utero and in the neonatal period. The successful management of AITD in pregnancy requires a knowledge of the differential placental transfer of various hormones, antibodies, and drugs. PMID- 18411135 TI - Newborn screening, prenatal diagnosis, and prenatal treatment of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency can be detected by newborn screening. Benefits of screening for affected newborns include the prevention of severe adrenal crisis, its sequelae, and progressive signs of androgen excess. First-trimester prenatal diagnosis is possible by HLA typing and/or DNA analysis of genes within the HLA complex of chorionic villus cells; for second-trimester diagnosis, hormonal measurement o f amniotic fluid and HLA typing or DNA analysis of amniotic cells are used. Results of prenatal treatment of CAH have varied, and the efficacy of prenatal treatment by maternal glucocorticoid therapy requires further investigation. PMID- 18411136 TI - Immune system and bone remodeling. AB - The skeleton undergoes continuous cellular remodeling in a manner similar to that of replacement of a wall, brick by brick. Products of immune cells are likely to modulate all of the cellular events involved in bone remodeling. The local factors that modulate bone cell function and bone remodeling-osteotropic cytokines or growth-regulatory factors-are similar to those involved in other body processes such as wound healing, immune function, oncogenesis, angiogenesis, and embryogenesis. PMID- 18411137 TI - The search for the elusive adrenal steroidogenic "regulatory" protein. AB - Inhibition of protein synthesis blocks ACTH-stimulated steroidogenesis. Researchers have proposed that a "regulatory" protein functions at an intramitochondrial site to promote the translocation of cholesterol substrate from the outer to the inner membrane where the rate-limiting P-450(scc) enzyme complex is located. To date three different proteins have been put forth as a "regulatory" protein. These are structurally distinct, yet appear to perform the same in vitro function; they are widely distributed, and are normally involved in disparate activities unrelated to steroidogenesis. Thus, it is difficult to understand how a physiological role for the three proteins in ACTH-stimulated steroidogenesis might exist. PMID- 18411138 TI - Growth hormone and the microvascular complications of diabetes. AB - The basis for the hypothesis that growth hormone is a permissive factor in the pathogenesis of diabetic microvascular complications is a weak one. The best way forward in this research will be to devise a pharmacological method of suppressing growth hormone secretion in diabetic subjects. PMID- 18411139 TI - Calcium and calmodulin regulation of prolactin gene expression. AB - Prolactin gene transcription is critically dependent upon intracellular calcium and the calcium-binding protein calmodulin. The effects of calcium and calmodulin on the prolactin gene appear to involve defined regions of the 5'-flanking sequence, and interactions between nuclear proteins and this DNA may be related to mechanisms of transcriptional control by pituitary cell-type-specific trans acting factors. PMID- 18411140 TI - Diabetes and hypertension: new concepts in treatment. AB - Since diabetes and hypertension frequently coexist in the same patient, it is important to consider that many antihypertensive agents may have diverse effects on carbohydrate metabolism, lipid profiles, and overall glycemic control, thus further confounding the management of patients with both diseases. The two cases below illustrate the potential metabolic effects of four commonly used antihypertensive medications (thiazide diuretics, beta-blockers, angiotensin converting enzyme inhibitors, and calcium channel blockers) and emphasize the complex interaction between these two common diseases and their treatments. PMID- 18411141 TI - Imaging techniques for identifying parathyroid tumors. AB - Recent advances in imaging techniques, including high-resolution sonography, parathyroid scintigraphy, computerized tomography, magnetic resonance imaging, venography and venous sampling, arteriography, and fine-needle aspiration biopsy, have made it possible consistently to identify parathyroid glands enlarged owing to adenoma formation, diffuse hyperplasia, or carcinoma. Whereas the routine use of these techniques in the previously unoperated patient with suspected hyperparathyroidism is debated, most experts would agree to their utility in patients with prior negative neck exploration in whom elevated serum calcium and parathyroid hormone persist. PMID- 18411142 TI - Growth hormone-binding proteins. AB - Two growth hormone-binding proteins (GH-BPs) have recently been discovered in human plasma. Both are specific for human GH; one has high affinity and the other has low affinity. The high-affinity BP corresponds to the extracellular portion of the GH receptor, whereas the low-affinity BP does not appear to be related to the receptor. About 45% of GH in plasma is complexed with the high-affinity BP, and another 5% with the low-affinity BP. Plasma BP levels are low in fetal life and rise in the first few years after birth to achieve adult levels, which stay constant. Some GH-resistant forms of short stature (Laron dwarfism, pygmies) are associated with absent, nonfunctional, or decreased levels of high-affinity BP in plasma. The BPs prolong the half-life of GH by confining its distribution and protecting it from degradation; they dampen oscillations and contribute to a circulating hormone reservoir/buffer. Certain animals, in particular the rabbit, pig, and pregnant mouse, possess similar GH-BPs. PMID- 18411143 TI - Modulation of hormone response elements by promoter environment. AB - Hormone response elements (HREs) are nucleotide sequences that confer onto promoters the ability to alter their transcriptional pace in response to hormones. Growing evidence indicates that the functional activity of HREs can be significantly modulated by their promoter environment, making it possible for genes containing the same HRE to display diversity in their responsiveness to a given hormone signal. PMID- 18411144 TI - The thyroglobulin gene: evolutionary and regulatory issues. AB - The coding information for thyroglobulin synthesis is contained in a large transcription unit, which is made from the juxtaposition of short repetitive gene modules and of a copy of an ancient acetylcholinesterase homologue. Minor alternatively spliced transcripts with conservation of the reading frame seem to be common. Whether they have a role or represent noise in the splicing phenomena is unknown. Transcription of the thyroglobulin gene is controlled by cAMP through a pathway involving sequence motifs and trans-acting factors that differ from those identified so far in other systems. PMID- 18411145 TI - Ontogeny of pituitary-gonadal interactions: current advances and controversies. AB - The different compartments of the fetal hypothalamic-pituitary-gonadal axis, the hypothalamus, anterior pituitary, and gonads, probably start their embryonic development independently, and become fully interactive as the last link o f their maturation. The developing hypothalamic-pituitary-gonadal axis offers a good model for studies on the mechanisms of regulation of fetal hormonal systems. It is evident that fetal hormonal functions are not the same as those of the adult on a smaller scale, but that there are fundamental differences between the fetus and adult in basic features of the mechanisms o f reproductive hormone action. PMID- 18411146 TI - Signal transduction processes leading to acrosomal exocytosis in mammalian spermatozoa. AB - Many components of intercellular signaling involved in species-specific sperm binding to the egg's extracellular matrix, the zona pellucida, and the induction of acrosomal exocytosis, an absolute prerequisite to successful fertilization, have properties similar to intercellular signaling mechanisms controlling somatic cell function. Sperm-associated receptors for zona pellucida glycoproteins have been postulated to serve as the initial components of signal transduction cascades leading to the stimulation of cellular effector systems that modulate sperm function. Such receptor-effector systems appear to be coupled through guanine nucleotide-binding regulatory proteins (G proteins). PMID- 18411147 TI - Immunosuppressive treatment of Graves' ophthalmopathy. AB - Steroids or orbital irradiation are effective in approximately 60% of patients with severe Graves' ophthalmopathy; the improvement of proptosis and extraocular muscle dysfunction, however, is limited. Better results are obtained by the combination of orbital irradiation and systemic steroids. Cyclosporine alone is not very effective, but the combination of cyclosporine and low-dose prednisone (administered after a previous insufficient response to high-dose steroids) is effective in 59% of cases. Disease activity rather than disease duration is probably the main determinant of the therapeutic outcome. PMID- 18411148 TI - Clinically useful and readily available techniques for measurements of bone mineral and body composition by photon or x-ray absorptiometry. AB - Technical develoments in dual-energy x-ray absorptiometry permit the estimation of bone mass or density and of body fat with one single, quick, nontraumatic measurement. Bone density is the best available means of estimating skeletal fracture risk. Improved fat quantification can be made independent from the generally made assumption in such calculations that bone represents a fixed fraction of fat-free body mass. PMID- 18411149 TI - The tissue-specific processing of Pro-ACTH/Endorphin recent advances and unsolved problems. AB - Pro-ACTH/endorphin (also called proopiomelanocortin) synthesis in corticotropes and melanotropes continues to serve as a model for investigating the series of enzymatic steps that convert inactive preprohormones to product peptides during transit of the molecules from the rough endoplasmic reticulum through the Golgi and into secretory granules. The complexity of the biosynthesis of bioactive peptides is increased by the widespread occurrence of tissue-specific and developmentally regulated posttranslational processing. Transfection of cDNAs encoding wild-type and mutant preproneuropeptide Y into AtT-20 corticotrope tumor cells revealed the importance of primary sequence in determining the extent of cleavage of the peptide precursor. The purification and cloning of several peptide-processing enzymes, including KEX-1, KEX-2, carboxypeptidase E, and peptidyl-glycine alpha-amidating monooxygenase, have provided important information about intracellular peptide processing, and are beginning to provide information about the sorting of soluble and membrane-associated components of secretory granules. PMID- 18411150 TI - beta-Core chemical and clinical properties. AB - beta-Core is the most abundant hCG-related molecule in pregnancy urine. The structure of beta-core as well as aspects of its metabolic clearance suggest that beta-core is a metabolic fragment of the hCG-beta subunit. The occurrence of beta core in the urine of patients with a broad spectrum of malignancies imparts an important role to beta-core as a tumor marker. The recent development of antisera with enhanced specificity and sensitivity for beta-core will facilitate further studies on the clinical significance of this molecule. PMID- 18411151 TI - Regulatory peptides produced in the anterior pituitary. AB - Several peptide families have been detected in the anterior pituitary of several species, including man, and for many of them evidence for local synthesis has been found. Although a paracrine action seems evident for a few, the precise function of most of these peptides remains unknown. PMID- 18411152 TI - Hypothalamic transplantation repair of reproductive defects in hypogonadal mice. AB - The defect of the hypogonadal mouse, resulting in infantile reproductive organs and severely reduced gonadotropin levels, is due to a truncation of the gene encoding for preprogonadotropinreleasing hormone. The hypogonadal mouse bearing a graft containing normal gonadotropin-releasing hormone neurons may show testicular development, seminal vesicle growth, and increased gonadotropin production in males. Normalization of gonadotropin levels in females is frequently associated with the capacity for a reflex ovulation followed by pregnancy and bearing of live young. All of these phenomena are dependent on the outgrowth of gonadotropin-releasing hormone axons from the graft to the host median eminence and the hypophysial portal capillaries. PMID- 18411153 TI - The role of immunopeptides in the regulation of anterior pituitary hormone release. AB - The anterior pituitary lobe secretes hormones that regulate the functioning of the immune system which, in turn, produces thymic hormones and interleukin proteins capable of altering neuroendocrine responsiveness. Interleukin-1 is released during inflammation and activates the hypothalamic-pituitary-adrenal axis, which subsequently diminishes the immune response. Interleukin-6 (IL-6) stimulates prolactin and growth hormone release in vitro from anterior pituitary cells which, in turn, are capable of producing IL-6. The possible production of IL-6 by the anterior pituitary in situ suggests an autocrine and/or paracrine role for this cytokine in the regulation of hormone release. PMID- 18411154 TI - Insulinlike growth factor binding proteins. AB - Insulinlike growth factor binding proteins (IGFBPs), which are often present in extracellular fluids, have the potential to markedly alter IGF-receptor interaction. There are at least four specific forms of IGFBPs in blood. These forms are also secreted by a number of cell types. Their rates of secretion and clearance appear to be regulated, mostly by hormones. Their major role seems to be to alter the interaction of IGFs with their cell surface receptors. Other functions such as transport out of the vasculature and modulating target cell actions also appear to be important. PMID- 18411155 TI - Chorionic gonadotropin secretion in normal, nonpregnant humans. AB - A chorionic gonadotropinlike material is present in blood and urine of normal, nonpregnant humans, is secreted in pulsatile fashion in parallel with luteinizing hormone, is stimulated by gonadotropin-releasing hormone, and is suppressed by GnRH agonists and estrogen. The source is probably the pituitary gland. PMID- 18411156 TI - Measurement of parathyroid hormone-related peptide in the circulation. AB - Newly developed assays for the measurement of circulating levels of parathyroid hormone-related peptide may become useful in distinguishing the syndrome of humoral hypercalcemia of malignancy from hypercalcemia of other etiologies. PMID- 18411157 TI - Hyperthyroidism due to Graves' disease in aged patients. AB - Hyperthyroidism due to Graves' disease is more prominent in older patients than was originally thought. Aged patients with this disorder have characteristic clinical and hormonal features that are different from those in younger patients. Thus, Graves' disease in aged patients is not merely a milder form of the disease seen in youth, but is qualitatively different in many aspects. Owing in part to incomplete understanding of the etiology of Graves' disease, the pathophysiologic reasons for these differences are unknown. PMID- 18411158 TI - Insulin in the brain what is its role? AB - Insulin and insulin receptors are present in brain and appear to function outside the realm of glucose homeostasis. Insulin receptors in the brain are structurally unique and appear to be found exclusively on neurons. Evidence from a variety of experimental systems indicates that insulin plays a role in neuronal growth and differentiation; additional functions may exist in the adult brain. Insulin action in neurons may be associated with unique signal transduction pathways. PMID- 18411159 TI - Pineal gland interface between the photoperiodic environment and the endocrine system. AB - The photoperiodic message that the pineal gland conveys to the organism is encoded in the circadian melatonin rhythm. Melatonin is a ubiquitously acting hormone that mediates seasonal changes in reproduction in nonhuman mammals and may have reproductive consequences in humans as well. Additionally, melatonin may relate to the function o f the immune system, hormone-responsive tumor growth, circadian rhythm disturbances, and a number of other processes. PMID- 18411160 TI - The impact of gene mapping techniques on the management of multiple endocrine neoplasia type 2. AB - There has been sustained progress toward the identification of the gene for multiple endocrine neoplasia type 2. Closely linked and flanking DNA markers have been identified, and it is now possible to assign gene carrier status in informative families at risk with a >90% certainty by the use of molecular genetic techniques. Application of-these techniques, however, requires an understanding of their current limitations and caution in their use of clinical decision making. PMID- 18411161 TI - Neuroendocrine peptides in milk. AB - Milk, which is a mammal-specific biologic fluid, contains several neuroendocrine peptides at concentrations higher than those found in plasma. These neuroendocrine peptides can be synthesized or processed in the mammary gland or excreted into milk through various pathways. In addition, certain milk proteins, notably casein, can be enzymatically processed to release "exorphins," peptides with opioid activities. In suckling mammals, hormones and neuropeptides are absorbed through the gastrointestinal tract and appear intact in the plasma. This absorption is age dependent and could have physiologic significance in neonatal development. PMID- 18411163 TI - Molecular basis of Laron dwarfism. AB - An autosomal recessive disorder, Laron-type dwarfism, results from peripheral unresponsiveness to growth hormone. Mutations in the growth hormone receptor have recently been identified in this syndrome. Analysis of patients with Laron-type dwarfism should provide insight into the mechanisms of hormone receptor binding and signal transduction pathways of this receptor, which belongs to a new class of transmembrane receptors. PMID- 18411162 TI - Intrathyroidal iodine heterogeneity of iodocompounds and kinetic compartmentalization. AB - In the normal thyroid, but not necessarily in the goitrous gland, the bulk of iodine is bound to thyroglobulin. Even in the normal thyroid-and much more so in goiters-iodine is contained in many different compartments with widely differing kinetics, biochemical composition, localization, and physiologic significance. Any change of thyroid function profoundly affects intrathyroidal iodine kinetics and produces a redistribution of stored iodine. This must be taken into account whenever the impact of a global change in intrathyroidal iodine stores on thyroid function and growth is studied in vivo. PMID- 18411164 TI - Adrenoleukodystrophy presenting as addison's disease in children and adults. AB - X-linked adrenoleukodystrophy (ALD) is a peroxisomal genetic disorder that causes adrenal insufficiency, demyelination in the central nervous system, and increased levels of very long chain fatty acids in tissues and body fluids. Although most cases appear in childhood as a devastating degenerative disorder or in adulthood as a milder disorder affecting the spinal cord, many patients have adrenal insufficiency prior to the onset of their neurologic deterioration for many years. Addison's disease without neurologic involvement may also remain the only clinical manifestation of X-linked ALD. Because of the prognostic implications, the need for genetic counseling, and the potential benefit of therapeutic interventions, any boy or adult male with Addison's disease must be tested for X linked ALD. PMID- 18411165 TI - Skeletal aging understanding the functional and structural basis of osteoporosis. AB - Recent years have seen considerable progress in our understanding of the normal processes by which peak adult bone mass is acquired and maintained, the physical, nutritional, and hormonal factors that regulate bone mass throughout adult life, and the critical role of reproductive hormonal replacement for maintaining bone mass within the early years after menopause. It is now apparent that loss of bone quantity is not sufficient to explain increased fracture risk. Bone fragility depends not only on its material properties, of which mineral content is an important component, but also on its microarchitecture and geometry. Age-related trabecular bone loss is manifest not simply as a global loss of bone, but is characterized by a loss of trabecular connectivity and increased cortical porosity. Appreciation of these issues along with understanding the lifelong process of bone remodeling are central to formulating rational strategies for preventing and treating osteoporosis. PMID- 18411166 TI - Osteoporosis Current practice and future perspectives. AB - Initiation of estrogens or other drugs as preventive measures for osteoporosis should be based upon objective estimates of actual, future fracture risk. Bone mass measurements, when considered in the context of age, life expectancy, expected bone loss, and other risk factors, enable improved patient risk stratification, and more rational treatment choices. PMID- 18411167 TI - Steroid hormones are multifunctional messengers to the brain. AB - Because the brain regulates the endocrine system and also responds to it, the changing environment as well as the experiences of the animal can change and shape properties of hormone-sensitive nerve cells. Depending upon the physiologic status of the organism, the effects of actions of steroid hormones can be trophic or damaging to the nervous system. Future research on the actions of steroids on the brain may advance the notion that the endocrine system has some responsibility for determining the individual traits of an animal in brain function and behavior. PMID- 18411168 TI - Modulation of IgA synthesis by neuroendocrine peptides. AB - Prolactin, estrogen, and progesterone together induce proliferation of the mammary glands with a coincident increase in the IgA-secreting plasma cells in the tissue. Of these three hormones, Prolactin is the most effective single peptide in stimulating IgA production. Vasoactive intestinal peptide (VIP) and somatostatin have also been shown to modulate IgA production. Although more extensive investigation is required, delineation of an immune axis involving prolactin, VIP, and somatostatin in mucosal immune tissue such as mammary gland is a promising area of research with clinical relevance to neonatal resistance to infection. PMID- 18411169 TI - Hormones and breast cancer. AB - The study of steroid hormone receptors in human breast cancer provides insight into tumor biology and forms the rationale for many therapeutic modalities. Clinical assays for estrogen and progesterone receptors improve the selection of patients who may benefit from hormonal therapy. In addition, receptor content in breast cancer tissue is useful for determining prognosis. Together with other new prognostic factors, hormone receptors help to distinguish among patients with primary breast cancer who are at risk for early recurrence. PMID- 18411170 TI - Nutritional rickets. AB - Nutritional rickets was once felt to be a disease of the past. In recent years, dietary patterns and the prevalence of unsupplemented breast feeding has led to a mild resurgence in rickets. The cases presented show a wide spectrum of nutritional disease, illustrating the common risk factors, the usefulness of vitamin-D metabolite assays in making a definitive diagnosis, and the response to vitamin-D therapy. PMID- 18411171 TI - Gonadotropin-releasing hormone agonist testing of pituitary-gonadal function. AB - The development of gonadotropin-releasing hormone (GnRH) agonists has provided a unique means to functionally assess the pituitary-gonadal axis in both males and females. These agonists, when given in a dose sufficient to stimulate the gonadotropes and induce a gonadal steroid response, have provided insights into normal reproductive physiology, hyperandrogenic conditions such as the polycystic ovary syndrome (PCOS), and disorders of pubertal development. This review provides an overview of the use of such agonists as probes of the functional status of the pituitary-gonadal axis in both normal and abnormal reproductive states. PMID- 18411172 TI - Contributions of growth hormone receptor and postreceptor defects to growth hormone resistance in malnutrition. AB - Malnutrition results in poor growth and is associated with resistance to growth hormone (GH) action. The mechanisms involved in the GH resistance depend on the severity and the timing of the nutritional insult. Stringent dietary restrictions such as fasting may produce GH resistance by reducing the number of GH receptors. Less severe nutritional deprivation such a short-term protein restriction may cause GH insensitivity mainly through postreceptor mechanisms. PMID- 18411173 TI - Transgenic mice for the study of diabetes mellitus. AB - Several recent studies have utilized transgenic technology to explore basic questions in the pathophysiology of diabetes mellitus. The ultimate expression of altered glucose homeostasis is a theme common to them. The experimental models have been diverse, however, and, in some instances, resulted in unexpected biologic effects. Many of the studies have examined the autoimmune etiology of insulin-dependent diabetes mellitus by expressing regulatory molecules of the immune system as transgenes in islet beta cells. The molecules have included products of the major histocompatibility complex (MHC), cytokines, and other cell surface antigens. Ectopic expression of these transgenes resulted in altered immune responses directed against islets, and these transgenic mice now serve as important models to study mechanisms of immunologic tolerance. Transgenic technology is also being used to explore basic aspects of islet beta-cell physiology and insulin metabolism. beta-cell function is disrupted by transgenic beta-cell expression of molecules such as calmodulin and H-ras. Hyperexpression of insulin as a transgene can result in a syndrome resembling features of non insulin-dependent diabetes. PMID- 18411174 TI - The renin-angiotensin system in the pituitary gland. AB - There is evidence that angiotensin II, in addition to being generated in the circulating blood, is synthesized in the anterior pituitary lobe and other endocrine tissues. Angiotensin II produced locally may act on pituitary cell receptors to modulate or mediate the action of other hormonal factors. However, tissue angiotensins may be synthesized by a different mechanism than most other neuroendocrine peptides. A precise understanding of the mode of formation of local angiotensin II is necessary for the comprehension of its physiologic role in the pituitary gland. PMID- 18411176 TI - Multipotential pituitary cells that contain adrenocorticotropin (ACTH) and other pituitary hormones. AB - Anterior pituitary cells that contain adrenocorticotropin (ACTH) are morphologically and functionally heterogeneous. Immunolabeling has identified stellate subtypes with scattered or peripherally arranged granules, and ovoid or irregularly shaped cells with a denser accumulation of granules, which overlap morphologically with other cell types such as small gonadotropes or thyrotropes. Dual cytochemical labeling studies on the same or serial fields have identified cells that store ACTH with LH, FSH, TSH, or prolactin. Evidence is presented to suggest that they are multipotential cells with the capacity to augment corticotropes or one of the other cell populations. PMID- 18411175 TI - Vasoactive intestinal polypeptide: biologic role in health and disease. AB - Vasoactive intestinal polypeptide (VIP), a neuropeptide with wide distribution in the central and peripheral nervous systems, has a broad spectrum of biologic actions. Usually acting as a neurotransmitter or neuromodulator but sometimes also as a blood-borne hormone, it participates in the regulation of a variety of major body functions and may be an important factor in the pathogenesis of several diseases. PMID- 18411177 TI - The modern management of macroprolactinomas. AB - Macroprolactinomas will usually shrink with dopamine agonist therapy, often to within the pituitary fossa; definitive treatment with radiotherapy can then achieve eventual normalization of serum prolactin in the majority of patients. Persistent suprasellar extensions, even when relatively large, can usually be successfully decompressed by the transsphenoidal route. Stalk compression can also give rise to hyperprolactinemia that will normalize with bromocriptine, but tumors causing it will rarely, if ever, show significant shrinkage in response to the drug. PMID- 18411178 TI - The polymerase chain reaction and its uses in endocrinology. AB - The polymerase chain reaction (PCR) represents a major technological development for molecular biologists and will have a great impact on molecular endocrinology. It is a novel technique that amplifies specific DNA sequences with remarkable efficiency. Repeated cycles of denaturation, primer annealing and extension carried out with the heat-stable enzyme, Taq polymerase, leads to exponential increases in the target DNA sequences. Because o f its sensitivity and specificity, this technique is being widely applied in all areas of molecular biology and human genetics. This article describes the basic PCR technique and explores its potential uses and applications to the field of endocrinology. PMID- 18411179 TI - Molecular biology of growth hormone receptors. AB - Growth hormone (GH), synthesized in and secreted from the anterior pituitary lobe, has multiple effects at both the cellular and the organismal levels, including promotion of body growth, stimulation of intermediary metabolism, and transcriptional regulation of specific genes. These effects are believed to be mediated by high-affinity cell-surface receptors, which have recently been cloned and characterized and found to be members of a newly described receptor family. Although GH receptor expression is widespread, the cellular mechanisms by which GH binding to those receptors initiates cellular responses are still unclear. Functional expression of the cloned receptor in tissue culture has resulted in substantial progress toward that goal. PMID- 18411180 TI - Regulation of pituitary gonadotropin gene expression outline of intracellular signaling pathways. AB - Genes encoding pituitary gonadotropins, luteinizing hormone, and follicle stimulating hormone, as well as their transcripts, have been isolated from different species and characterized. The position of introns in the genes is highly conserved in different species, and it appears that sequences encoding gonadotropin subunits evolved from an ancestral gene. Expression of gonadotropin genes is regulated mainly by gonadotropin-releasing hormone (GnRH) and by gonadal steroids (estradiol and testosterone). Other factors, notably thyroid hormones and gonadal peptides (follistatin, inhibin, and activin), also play a role in this regulation. Although the intracellular mechanisms of GnRH signal transduction are still unclear, now a body of evidence suggests the involvement of protein kinases C and A in intermediary steps of gonadotropin synthesis and release. PMID- 18411181 TI - The role of myo-inositol in the pathogenesis of diabetic complications. AB - It is tempting to speculate that all diabetic complications are generated by a single biochemical abnormality. The observation that intracellular inositol depletion occurs in diabetes mellitus has led to the hypothesis that this may be of importance in the pathogenesis of diabetic complications. The central role of inositol and inositol derivatives in cellular function has given support to this theory, but as yet no fully substantiated mechanistic model can be offered. PMID- 18411182 TI - Body weight, body fat, and ovulation. AB - Women with moderate weight loss (10%-15% of ideal weight) have secondary or primary amenorrhea. Athletic women also may have amenorrhea, or anovulatory cycles, though body weight may be in the normal range, since muscles are heavy (80% water compared with 5%-10% water in adipose tissue). The amenorrhea, which is usually reversible with weight gain, decreased exercise, or both, is due to hypothalamic dysfunction; the pituitary-ovary axis is intact, suggesting that this type of amenorrhea is adaptive, preventing an unsuccessful pregnancy. Evidence is presented that the high percentage of body fat (26%-28%) in mature women is necessary for regular ovulatory cycles and may influence reproductive ability directly: (a) as an extragonadal source of estrogen by aromatization of androgen to estrogen, (b) by influencing the direction of estrogen metabolism to more potent or less potent forms, or (c) by changes in the binding properties of sex-hormone-binding globulin. Indirect signals may be abnormal control of temperature and changes in energy metabolism that accompany excessive leanness. PMID- 18411183 TI - Islet amyloid polypeptide: the cause of type-2 diabetes? AB - Islet amyloid polypeptide (IAPP or amylin), first identified as the peptide deposited as amyloid in type-2 diabetic pancreas and insulinoma, turns out to be a peptide produced in the pancreatic beta-cell secretory granule that is costored and coreleased with insulin. Experimental evidence suggests that, under certain conditions, IAPP can counter insulin action in peripheral tissue and inhibit insulin release from the pancreas. IAPP therefore appears to respond to the same physiologic stimuli as insulin, but has opposing biologic actions. The role of IAPP, both in normal physiology and in pathology, remains unclear, but current evidence suggests against a role as a circulating hormone in favor of a paracrine or autocrine modulator of insulin secretion. PMID- 18411184 TI - Paget's disease of bone. AB - Paget's disease of bone is a common disorder of older patients that may sometimes cause a variety of signs and symptoms referable to the skeleton. An intrinsic bone-remodeling abnormality in this disease may lead to bone deformity, pain, and various arthritic or neurologic complications. Characteristic radiographic changes can be seen, and typically the serum alkaline phosphatase level and total urinary hydroxyproline excretion are elevated. Current therapies that interfere with excess osteoclast activity are helpful in many cases, but treatment choices and expected responses must be considered in the context of the individual patient. PMID- 18411185 TI - Tumor necrosis factor, cytokines, and the hyperlipidemia of infection. AB - The hypertriglyceridemia of infection is produced by many of the cytokines that regulate the immune response, including the tumor necrosis factors, interleukin 1, interleukin-6, and the interferons. In rodents, this hypertriglyceridemia is primarily due to the ability of these cytokines to stimulate hepatic lipogenesis and very low density lipoprotein production without causing significant changes in adipose tissue lipoprotein lipase or triglyceride clearance. Induction of hypertriglyceridemia is not directly linked to the syndrome of wasting. Rather, the increase in hepatic production of lipoproteins is part of the acute-phase response and may contribute to host defense. PMID- 18411186 TI - Current status of brain hypophysiotropic factors: morphologic aspects. AB - Nearly 40 putative neurotransmitters and other chemical messengers, mostly peptides, are present in the median eminence that constitutes the final common pathway for signals from the brain to the pituitary. The majority of them are produced in perikarya located in different nuclei of the hypothalamus; however, some of them arise from the brainstem. The neurons contacting capillaries of the median eminence (hypophysiotropic neurons) are intermixed with neurons containing the same transmitter (hypophysiotropic factor(1)), but projecting to other areas of the brain. Depending on their site of release, the hypophysiotropic factors may function as neurohormones acting an the pituitary or neurotransmitters affecting the activity of other neurons in the central nervous system. Based on retrograde tracing studies in combination with immunocytochemistry, the origin of many nerve terminals in the median eminence has been determined. PMID- 18411187 TI - Anti-Mullerian hormone and intersex states. AB - Anti-Mullerian hormone (AMH), alias Mullerian-inhibiting substance or factor, plays a key role in fetal sex differentiation. The cloning of the human gene, a member of the transforming growth-factor-beta family and the development of immunochemical reagents recognizing circulating human AMH have opened new perspectives for clinical research. AMH assays and genetic studies now provide meaningful information regarding testicular function in infancy and the molecular basis of a rare form of male pseudohermaphroditism, the persistent Mullerian duct syndrome. PMID- 18411188 TI - The antisteroid RU486: its cellular and molecular mode of action. AB - RU486 is a very powerful antisteroid hormone compound, with antiprogestin and antiglucocorticosteroid clinical applications. High-affinity binding to the receptor ligand-binding domain results in essentially antagonistic function; however, some agonistlike effects have been observed. This review discusses the implications of recent discoveries concerning hsp90 (heat-shock protein M(r) approximately 90,000)-receptor interaction, receptor-DNA binding, and the involvement of transcription factors in the mechanism of action of RU486. PMID- 18411189 TI - ASEICA's 25th anniversary. PMID- 18411190 TI - An interview with Purificacion Garcia de Miguel, chief of Haematology-Oncology and the Paediatric Bone Marrow Transplantation Unit of the Hospital La Paz, Madrid, and President of the Spanish Society of Paediatric Oncology (SEOP). Interview. PMID- 18411191 TI - Genetic changes in small cell lung carcinoma. AB - Small cell lung carcinoma (SCLC) accounts for approximately 15% of all lung cancer cases. Despite a frequently good response to first-line treatment with chemotherapy and/or radiotherapy, early relapse occurs in the majority of patients and 5-year survival is only about 5%. Therefore, there is a need to develop novel treatments to improve the outcome of patients with SCLC. To fulfil this need, it is critical to gain further understanding on the molecular basis of SCLC and specifically to identify novel therapeutic targets. Clinical trials with molecularly targeted agents have been performed with little success in the past, but recently many promising oncogenic pathways have been discovered and novel targeted therapies are under evaluation. In this review, we summarise the most relevant genetic and signalling pathway alterations reported to date in SCLC and discuss the potential therapeutic implications of such events. PMID- 18411192 TI - Angiogenesis inhibitors in the treatment of non-small-cell lung cancer (NSCLC). AB - Lung cancer is a leading cause of death worldwide and, although some progress has been made in its treatment, the results remain poor. Better knowledge in tumour biology has allowed us to design anti-target drugs and incorporate them in the treatment of non-small-cell lung cancer (NSCLC). One of the most widely used targeted approaches in this type of tumour has been the inhibition of angiogenesis. Several strategies blocking the VEGF pathway, either at the ligand or recepor level, have been studied and developed. In this review, we present an up-to-date analysis of the current inhibitors of angiogenesis in the treatment of NSCLC. PMID- 18411193 TI - On the relevance of defining protein structures in cancer research. AB - Are three-dimensional structures of proteins relevant in the study of cancer? The knowledge of the three-dimensional structure of a protein is crucial to gain a full understanding of its function, and structural determination has already shown its potential for guided drug design. The knowledge of the structures of proteins and their complexes with other biological macromolecules helps to elucidate functional networks and provide a better understanding of the functionally relevant behaviour of the molecular machinery of the cell. To study the cell, we must be able to work with proteins, to elucidate how they diffuse and move, to know their interacting partners, and to understand the changes induced by those interactions. Three-dimensional structures give us a picture of the protein and thereby the opportunity to introduce mutations that alter its affinity and specificity for other interactions helping us to understand the physico-chemical mechanisms that control their function. In turn these can lead to the development of novel therapies. PMID- 18411194 TI - Implementation of a comprehensive cancer plan. A health planning experience. AB - Here we describe the working method used to implement an Integrated Cancer Health Plan in Andalusia (Spain) and to set out some of the lines of work for improving quality of care and health network planning. Four main initial considerations were made: (1) work must be centred on patients and respect for their autonomy; (2) cancer requires action at every healthcare level; (3) integration of expert professionals is to be encouraged; and (4) relevant information and data should be systematically used for planning. Two operative approaches were also established: structured participation and evidence-based healthcare. This methodology was used in various planning programmes, largely in relation to healthcare resources. In this article, as examples, we describe the development of two types of programmes: one for radiotherapy equipment and another for the management of lymphoedema in breast cancer patients. Analysis of results obtained against cancer is always controversial. It is necessary to review the healthcare structure used in terms of effectiveness and excellence. Comprehensive or integrated health plans are a useful model for a wide-ranging and multi-level approach to cancer. The structured and real participation of experts and an evidence- based healthcare strategy proved very useful in this public health planning experience. PMID- 18411195 TI - An easy, rapid and objective mathematical method to identify fatty acid synthase (oncogenic antigen-519) modulators with potential anticancer value. AB - Fatty acid synthase (FASN) is a novel druggable target for metabolically treating and preventing human malignancies. We envisioned that if loss of sensitivity to C75 (a slow-binding FASN inhibitor) occurs in parallel with loss of FASN expression and/or activity, a mathematical assessment of the nature of the interaction between investigational FASN modulators and C75 may predict the ability of experimental compounds to regulate FASN. We statistically compared the arithmetical sums of the anti-proliferative effects obtained when FASN modulators and C75 were used as single agents to those observed experimentally when agents were actually combined in a sequential schedule (i.e., FASN modulator-->C75). A reduced sensitivity to C75 (antagonism) occurred when compounds down-regulated FASN activity/expression, while an enhanced C75 efficacy (synergism) was found following exposure to FASN up-regulators. This "C75-sensitivity test" might offer an easy, rapid and objective method to identify FASN inhibitors with potential anticancer value in human cancer. PMID- 18411196 TI - Expression of blood group-related antigens in neoplastic uterine cervix. AB - Expression of blood group antigens in normal, displastic and tumoral uterine cervix from 35 hysterectomised women with carcinoma of the cervix was investigated; the results were correlated with patients' ABH phenotype and secretor status. We used an indirect immunoperoxidase technique and a panel of monoclonal antibodies and lectins directed against different antigenic specificities. Anomalous expression of blood group antigens in premalignant lesions from cervix was found. Partial loos of expression of blood group antigens and some lectins in different grades of cervical intraepithelial neoplasia, and a total loss of expression in CIN III and in infiltrating carcinoma of the cervix from secretor patients was revealed. The findings herein described confirm the importance of these antigens as tumour markers and they might be useful for the study of cervical carcinogenesis. PMID- 18411197 TI - Responsibilities in cancer preventive care in Greece. A physicians' survey. AB - BACKGROUND: Colorectal cancer is the second leading cause of cancer death in European countries. Differences in screening implementation may explain USA vs. European survival differences. The proportion of European primary care physicians advising colorectal screening has been reported to be inconsistent. We therefore hypothesised the presence of a belief-related bias among European physicians regarding who is responsible for cancer screening delivery. OBJECTIVES: To index beliefs in cancer screening implementation among a wide sample of Greek physicians. Study design Cross-sectional survey. METHODS: Three hundred and sixty six physicians involved in primary care activities in 15 provinces answered a questionnaire about responsibility in cancer screening delivery. Results 22.4% and 7.6% of physicians declared that the health system and the patients, respectively, have the main responsibility for cancer screening implementation, while 70 % advocated patient-health system co-responsibility. Beliefs were statistically correlated to age (p=0.039) and specialisation category (p=0.002). Patients' will was mainly indicated by internists, trainee internists and physicians older than 30, while GPs, trainee GPs and house officers were mainly health system-oriented. Worryingly, when physicians were asked about which specialty should inform the population, 81% indicated family doctor (for-fee service) while the involvement of free-from-fee specialities was inconsistent. CONCLUSION: A considerable disorientation about responsibilities in cancer screening delivery was observed in our study sample. Continual medical education and clear redefinition of primary care physicians' activities are required. PMID- 18411198 TI - Megestrol acetate-induced adrenal insufficiency. AB - Megestrol acetate is a synthetic progestin that has been used since the 1970s for the treatment of advanced cancer and subsequently to treat anorexia, cachexia and weight loss in AIDS patients. It has been shown that high doses or prolonged treatment with this drug may cause Cushing's syndrome, new-onset diabetes and suppression of plasma ACTH and cortisol levels. Megestrol acetate may cause suppression of the pituitary-adrenal axis due to the affinity of this compound for the glucocorticoid receptor. Recognising the glucocorticoid-like activity of megestrol and its effects at the axis level is important for the diagnosis of sub clinical adrenal insufficiency. We present the case of a 74-year-old woman with infiltrating ductal breast carcinoma refractory to prolonged hormonal treatment with megestrol acetate, presenting with adrenal insufficiency. PMID- 18411199 TI - Retroperitoneal Castleman's disease with colon cancer. A rare association. AB - Castleman's disease (CD) is a rare disorder of uncertain aetiology characterised by massive proliferation of lymphoid tissue usually localised as mediastinal masses, although abdominal involvement has been reported. Localised forms are usually associated with a good prognosis, but several more aggressive multifocal variants have been observed. Two different histologic subtypes have been described: the hyaline vascular type, more common in unicentric CD and usually asymptomatic, and the plasma cell form. Unicentric CD may be associated with an increased risk of lymphoma, but there was no reported increased risk of other malignancies. A patient with plasma cell subtype unicentric CD localised in retroperitoneum associated with an adenocarcinoma of ileocaecal valve and liver metastasis is reported. PMID- 18411200 TI - Multiple bone lesions resembling a metastatic origin. An unexpected diagnosis. AB - Lytic and blastic lesions have been associated to malignant tumours, such as solid cancer (breast cancer, renal cancer, prostate cancer, malignant melanoma or thyroid tumours). Although a mixed pattern with lytic and blastic lesions is due to metastatic tumour, this is not the only possible origin. The following case shows a systematic. This case report shows the number of tests that were made in order to discover the origin of osteolytic and osteoblastic lesions and it is notable that there is not an occult neoplasia on every occasion. PMID- 18411201 TI - Out-of-pocket health expenditure in a population covered by the Family Health Program in Brazil. AB - BACKGROUND: The Brazilian public health system, free and universal, should limit out-of-pocket health expenses. However, Brazil was reported as one of the countries with the highest proportion of families experiencing catastrophic expenditure. This study was designed to assess occurrence of high health spending in a low-income population, as well as the pattern of out-of-pocket health payments. METHODS: A cross-sectional study was done in Porto Alegre, Brazil, in 2003, with a sample representative of families covered by the Family Health Program. Health expenses were recorded with reference to 30 days prior to the interview and income data were collected with reference to the previous calendar month. Health expenditure was explored in terms of total household health expenditure >5, 10 and 20% of household income and >40% households capacity to pay. RESULTS: The final study sample included 869 households. Medicines were responsible for 47% of household expenditure with health; second came private health plans which accounted for 22%. The richest spent, on average, 70 times more them the poorest with health plans, 26 times more with dental treatment and six times more with medicines. About 16% households committed 20% or more of their income with health, independent of economic position. Similarly, 12% of the households had health expenditure in excess of 40% of their capacity to pay. CONCLUSION: The proportion of income spent on health was similar across economic groups, but this 'equality' is achieved at an unacceptably high level. Specific strategies to reduce such vulnerability are needed. PMID- 18411202 TI - The CGView Server: a comparative genomics tool for circular genomes. AB - The CGView Server generates graphical maps of circular genomes that show sequence features, base composition plots, analysis results and sequence similarity plots. Sequences can be supplied in raw, FASTA, GenBank or EMBL format. Additional feature or analysis information can be submitted in the form of GFF (General Feature Format) files. The server uses BLAST to compare the primary sequence to up to three comparison genomes or sequence sets. The BLAST results and feature information are converted to a graphical map showing the entire sequence, or an expanded and more detailed view of a region of interest. Several options are included to control which types of features are displayed and how the features are drawn. The CGView Server can be used to visualize features associated with any bacterial, plasmid, chloroplast or mitochondrial genome, and can aid in the identification of conserved genome segments, instances of horizontal gene transfer, and differences in gene copy number. Because a collection of sequences can be used in place of a comparison genome, maps can also be used to visualize regions of a known genome covered by newly obtained sequence reads. The CGView Server can be accessed at http://stothard.afns.ualberta.ca/cgview_server/ PMID- 18411203 TI - DAhunter: a web-based server that identifies homologous proteins by comparing domain architecture. AB - We present DAhunter, a web-based server that identifies homologous proteins by comparing domain architectures, the organization of protein domains. A major obstacle in comparison of domain architecture is the existence of 'promiscuous' domains, which carry out auxiliary functions and appear in many unrelated proteins. To distinguish these promiscuous domains from protein domains, we assigned a weight score to each domain extracted from RefSeq proteins, based on its abundance and versatility. A domain's score represents its importance in the 'protein world' and is used in the comparison of domain architectures. In scoring domains, DAhunter also considers domain combinations as well as single domains. To measure the similarity of two domain architectures, we developed several methods that are based on algorithms used in information retrieval (the cosine similarity, the Goodman-Kruskal gamma function, and domain duplication index) and then combined these into a similarity score. Compared with other domain architecture algorithms, DAhunter is better at identifying homology. The server is available at http://www.dahunter.kr and http://localodom.kobic.re.kr/dahunter/index.htm. PMID- 18411204 TI - Accuracy and application of the motif expression decomposition method in dissecting transcriptional regulation. AB - Understanding transcriptional regulation is a major goal of molecular biology. Motif expression decomposition (MED) was recently introduced to describe the expression level of a gene as the sum of the products of the binding strengths of its cis-regulatory motifs and the activities of the corresponding trans-acting transcription factors (TFs). Here, we use computer simulation to examine the accuracy of MED. We found that although MED accurately rebuilds gene expression levels from decomposed motif binding strengths and TF activities, estimates of motif binding strengths and TF activities are unreliable. Nonetheless, MED provides accurate estimates of relative binding strengths of the same motif in different genes and relative activities of the same TF under different conditions. We found that reasonably accurate results are achievable with genome wide expression data from only 30 conditions and that MED results are robust to the existence of unknown occurrences of known motifs, although they are less robust to the presence of unknown motifs. With these understandings, judicious use of MED will likely provide useful information about eukaryotic transcriptional regulation. As an example, MED results are used to demonstrate that motifs generally have higher binding strengths when appearing in multiple copies than appearing in one copy per promoter. PMID- 18411205 TI - DNA bar coding and pyrosequencing to analyze adverse events in therapeutic gene transfer. AB - Gene transfer has been used to correct inherited immunodeficiencies, but in several patients integration of therapeutic retroviral vectors activated proto oncogenes and caused leukemia. Here, we describe improved methods for characterizing integration site populations from gene transfer studies using DNA bar coding and pyrosequencing. We characterized 160,232 integration site sequences in 28 tissue samples from eight mice, where Rag1 or Artemis deficiencies were corrected by introducing the missing gene with gamma-retroviral or lentiviral vectors. The integration sites were characterized for their genomic distributions, including proximity to proto-oncogenes. Several mice harbored abnormal lymphoproliferations following therapy--in these cases, comparison of the location and frequency of isolation of integration sites across multiple tissues helped clarify the contribution of specific proviruses to the adverse events. We also took advantage of the large number of pyrosequencing reads to show that recovery of integration sites can be highly biased by the use of restriction enzyme cleavage of genomic DNA, which is a limitation in all widely used methods, but describe improved approaches that take advantage of the power of pyrosequencing to overcome this problem. The methods described here should allow integration site populations from human gene therapy to be deeply characterized with spatial and temporal resolution. PMID- 18411206 TI - Genome level analysis of rice mRNA 3'-end processing signals and alternative polyadenylation. AB - The position of a poly(A) site of eukaryotic mRNA is determined by sequence signals in pre-mRNA and a group of polyadenylation factors. To reveal rice poly(A) signals at a genome level, we constructed a dataset of 55 742 authenticated poly(A) sites and characterized the poly(A) signals. This resulted in identifying the typical tripartite cis-elements, including FUE, NUE and CE, as previously observed in Arabidopsis. The average size of the 3'-UTR was 289 nucleotides. When mapped to the genome, however, 15% of these poly(A) sites were found to be located in the currently annotated intergenic regions. Moreover, an extensive alternative polyadenylation profile was evident where 50% of the genes analyzed had more than one unique poly(A) site (excluding microheterogeneity sites), and 13% had four or more poly(A) sites. About 4% of the analyzed genes possessed alternative poly(A) sites at their introns, 5'-UTRs, or protein coding regions. The authenticity of these alternative poly(A) sites was partially confirmed using MPSS data. Analysis of nucleotide profile and signal patterns indicated that there may be a different set of poly(A) signals for those poly(A) sites found in the coding regions. Based on the features of rice poly(A) signals, an updated algorithm termed PASS-Rice was designed to predict poly(A) sites. PMID- 18411207 TI - Uncovering signal transduction networks from high-throughput data by integer linear programming. AB - Signal transduction is an important process that transmits signals from the outside of a cell to the inside to mediate sophisticated biological responses. Effective computational models to unravel such a process by taking advantage of high-throughput genomic and proteomic data are needed to understand the essential mechanisms underlying the signaling pathways. In this article, we propose a novel method for uncovering signal transduction networks (STNs) by integrating protein interaction with gene expression data. Specifically, we formulate STN identification problem as an integer linear programming (ILP) model, which can be actually solved by a relaxed linear programming algorithm and is flexible for handling various prior information without any restriction on the network structures. The numerical results on yeast MAPK signaling pathways demonstrate that the proposed ILP model is able to uncover STNs or pathways in an efficient and accurate manner. In particular, the prediction results are found to be in high agreement with current biological knowledge and available information in literature. In addition, the proposed model is simple to be interpreted and easy to be implemented even for a large-scale system. PMID- 18411208 TI - Structure and function of the regulatory C-terminal HRDC domain from Deinococcus radiodurans RecQ. AB - RecQ helicases are critical for maintaining genome integrity in organisms ranging from bacteria to humans by participating in a complex network of DNA metabolic pathways. Their diverse cellular functions require specialization and coordination of multiple protein domains that integrate catalytic functions with DNA-protein and protein-protein interactions. The RecQ helicase from Deinococcus radiodurans (DrRecQ) is unusual among RecQ family members in that it has evolved to utilize three 'Helicase and RNaseD C-terminal' (HRDC) domains to regulate its activity. In this report, we describe the high-resolution structure of the C terminal-most HRDC domain of DrRecQ. The structure reveals unusual electrostatic surface features that distinguish it from other HRDC domains. Mutation of individual residues in these regions affects the DNA binding affinity of DrRecQ and its ability to unwind a partial duplex DNA substrate. Taken together, the results suggest the unusual electrostatic surface features of the DrRecQ HRDC domain may be important for inter-domain interactions that regulate structure specific DNA binding and help direct DrRecQ to specific recombination/repair sites. PMID- 18411209 TI - Copper sensing function of Drosophila metal-responsive transcription factor-1 is mediated by a tetranuclear Cu(I) cluster. AB - Drosophila melanogaster MTF-1 (dMTF-1) is a copper-responsive transcriptional activator that mediates resistance to Cu, as well as Zn and Cd. Here, we characterize a novel cysteine-rich domain which is crucial for sensing excess intracellular copper by dMTF-1. Transgenic flies expressing mutant dMTF-1 containing alanine substitutions of two, four or six cysteine residues within the sequence (547)CNCTNCKCDQTKSCHGGDC(565) are significantly or completely impaired in their ability to protect flies from copper toxicity and fail to up-regulate MtnA (metallothionein) expression in response to excess Cu. In contrast, these flies exhibit wild-type survival in response to copper deprivation thus revealing that the cysteine cluster domain is required only for sensing Cu load by dMTF-1. Parallel studies show that the isolated cysteine cluster domain is required to protect a copper-sensitive S. cerevisiae ace1Delta strain from copper toxicity. Cu(I) ligation by a Cys-rich domain peptide fragment drives the cooperative assembly of a polydentate [Cu(4)-S(6)] cage structure, characterized by a core of trigonally S(3) coordinated Cu(I) ions bound by bridging thiolate ligands. While reminiscent of Cu(4)-L(6) (L = ligand) tetranuclear clusters in copper regulatory transcription factors of yeast, the absence of significant sequence homology is consistent with convergent evolution of a sensing strategy particularly well suited for Cu(I). PMID- 18411211 TI - Prevalence and significance of an isolated long QT interval in elite athletes. PMID- 18411210 TI - CSI-Tree: a regression tree approach for modeling binding properties of DNA binding molecules based on cognate site identification (CSI) data. AB - The identification and characterization of binding sites of DNA-binding molecules, including transcription factors (TFs), is a critical problem at the interface of chemistry, biology and molecular medicine. The Cognate Site Identification (CSI) array is a high-throughput microarray platform for measuring comprehensive recognition profiles of DNA-binding molecules. This technique produces datasets that are useful not only for identifying binding sites of previously uncharacterized TFs but also for elucidating dependencies, both local and nonlocal, between the nucleotides at different positions of the recognition sites. We have developed a regression tree technique, CSI-Tree, for exploring the spectrum of binding sites of DNA-binding molecules. Our approach constructs regression trees utilizing the CSI data of unaligned sequences. The resulting model partitions the binding spectrum into homogeneous regions of position specific nucleotide effects. Each homogeneous partition is then summarized by a position weight matrix (PWM). Hence, the final outcome is a binding intensity rank-ordered collection of PWMs each of which spans a different region in the binding spectrum. Nodes of the regression tree depict the critical position/nucleotide combinations. We analyze the CSI data of the eukaryotic TF Nkx-2.5 and two engineered small molecule DNA ligands and obtain unique insights into their binding properties. The CSI tree for Nkx-2.5 reveals an interaction between two positions of the binding profile and elucidates how different nucleotide combinations at these two positions lead to different binding affinities. The CSI trees for the engineered DNA ligands exhibit a common preference for the dinucleotide AA in the first two positions, which is consistent with preference for a narrow and relatively flat minor groove. We carry out a reanalysis of these data with a mixture of PWMs approach. This approach is an advancement over the simple PWM model and accommodates position dependencies based on only sequence data. Our analysis indicates that the dependencies revealed by the CSI-Tree are challenging to discover without the actual binding intensities. Moreover, such a mixture model is highly sensitive to the number and length of the sequences analyzed. In contrast, CSI-Tree provides interpretable and concise summaries of the complete recognition profiles of DNA binding molecules by utilizing binding affinities. PMID- 18411212 TI - Successful treatment of rheumatoid vasculitis-associated cutaneous ulcers using rituximab in two patients with rheumatoid arthritis. PMID- 18411213 TI - Vitamin D deficiency in systemic lupus erythematosus: prevalence, predictors and clinical consequences. AB - OBJECTIVES: We aimed to establish the prevalence, predictors and clinical consequences of vitamin D deficiency in patients with SLE. METHODS: Cross sectional study including patients fulfilling ACR criteria for the classification of SLE. Serum 25(OH)D levels at 30 and 10 ng/ml were the cut-off values for vitamin D insufficiency and vitamin D deficiency, respectively. SLE activity was measured by SLEDAI and irreversible organ damage by the SLICC-ACR index. Fatigue was quantified using a 0-10 visual analogue scale (VAS). RESULTS: Ninety-two patients (90% women, 98% white) were included in the study. Sixty-nine (75%) and 14 (15%) patients presented with vitamin D insufficiency and deficiency, respectively. Female sex (P = 0.001), treatment with HCQ (P = 0.014) and treatment with calcium and vitamin D (P = 0.049) predicted higher levels of 25(OH)D. Photosensitivity [odds ratio (OR) 3.5] and photoprotection (OR 5.7) predicted vitamin D insufficiency and deficiency, respectively. Higher age (OR 0.95) and HCQ use (OR 0.29) protected against vitamin D deficiency. Patients with vitamin D deficiency had a higher degree of fatigue as quantified by a 0-10 VAS (mean 5.32 vs 4.03, P = 0.08). No relation was seen between vitamin D insufficiency or deficiency and disease duration, SLEDAI or SLICC-ACR indexes. CONCLUSIONS: Vitamin D insufficiency and deficiency are common in patients with SLE and are associated with sun avoidance. HCQ prevented vitamin D deficiency. Vitamin D deficiency was related to a higher degree of fatigue. Vitamin D levels had no relation with SLE severity. PMID- 18411214 TI - Autonomic nervous symptoms in primary Sjogren's syndrome. AB - OBJECTIVES: Objective signs of autonomic dysfunction (AD) have been reported in patients with primary SS (pSS) while the presence of associated symptoms has not been systematically studied. Therefore, the aims of this study were (i) to assess the presence and severity of various AD symptoms in pSS patients and (ii) to relate AD symptoms to other clinical features of pSS. METHODS: Thirty-eight pSS patients and 200 population-based controls were studied for presence and severity of AD symptoms using the Autonomic Symptom Profile (ASP), a validated self completed questionnaire evaluating various AD symptoms. In addition, patients were investigated by three different objective autonomic nervous function tests. RESULTS: pSS patients scored significantly higher in the parasympathetic [secretomotor disorder, urinary disorder, gastroparesis (females only) and pupillomotor disorder] as well as sympathetic (orthostatic intolerance and vasomotor disorder) ASP domains compared with controls. Consequently, the standardized ASP total score was significantly increased in pSS patients [1.77 (0.57, 3.15) vs - 0.21 (-0.82, 0.72); P = 0.00] and 45% of pSS patients had an ASP total score >/=2 s.d. Furthermore, the autonomic nervous function tests showed signs of objective parasympathetic and sympathetic dysfunction as well. However, the ASP domain and total scores showed limited associations with the objective autonomic nervous function test parameters as well as clinical and serological factors of pSS. CONCLUSIONS: pSS patients showed subjective and objective signs of both a parasympathetic and a sympathetic dysfunction. However, AD symptoms showed limited associations with objective autonomic nervous function as well as other clinical features of the disease. PMID- 18411216 TI - The UK Clinical Research Network--building a world-class infrastructure for clinical research. PMID- 18411215 TI - Bolus infusion of human urinary trypsin inhibitor improves intractable interstitial pneumonia in patients with connective tissue diseases. AB - OBJECTIVE: Interstitial pneumonia (IP) associated with CTDs often progresses despite conventional immunosuppressive treatment. We investigated the efficacy of human urinary trypsin (UT) inhibitor (ulinastatin) on refractory IP. METHODS: Five patients with IP received UT inhibitor (3 x 10(5) U) infusion into the internal jugular vein, three times in a single day. The response to this therapy was assessed clinically and by chest CT, PaO(2) and serum KL-6. The kinetics of UT inhibitor was determined in arterial blood. We measured serum levels of monocyte chemotactic protein-1 and TGF-beta1, which are thought to be involved in the pathogenesis of IP. RESULTS: Serum concentrations of UT inhibitor increased immediately to >150 U/ml after infusion of 3 x 10(5) U of UT inhibitor. The treatment resulted in clinical and radiological improvements in four patients, and allowed reduction of oxygen therapy following improvement of hypoxaemia within 1 month. UT inhibitor decreased serum levels of KL-6 in all patients and had no adverse effects. MCP-1 and TGF-beta1 concentrations were higher in the patients than in normal subjects, and infusion of 3 x 10(5) U of UT reduced the concentrations within 3 h of infusion. CONCLUSION: UT inhibitor bolus infusion therapy is a potentially useful therapeutic strategy for intractable IP based on the different mechanism of action relative to conventional immunosuppressive therapy and lack of serious treatment-related adverse effects. PMID- 18411217 TI - Effects of vitamin D on expression of Toll-like receptors of monocytes from patients with Behcet's disease. AB - OBJECTIVES: Recent studies have shown the immunomodulatory effect of vitamin D(3) through down-regulation of Toll-like receptor (TLR) expression in human monocytes. To understand the implication of innate immunity with the role of vitamin D affecting TLR expression in Behcet's disease (BD), we focused on the association between the TLR expression and the serum vitamin D concentration in BD. METHODS: The expression of TLR2, TLR4 and CD16 on monocytes was detected by flow cytometric analysis and RT-PCR. Serum 25-hydroxyvitamin D [25(OH)D] levels were measured in the patients with BD, psoriasis and healthy controls, and then the expression of TLRs was correlated with the value of serum 25(OH)D levels. To assess the influence of vitamin D(3) on expression and function of TLRs in vitro, human monocytes were treated with increasing concentrations of 1,25(OH)(2)D(3). RESULTS: We found that the monocytes of active BD patients showed higher expressions of TLR2 and TLR4 than those of controls, and serum 25(OH)D levels tended to be lower in active BD. Furthermore, 25(OH)D levels were inversely correlated with the expressions of TLR2, TLR4 and clinical indicators. In vitro analysis showed that vitamin D(3) was found to dose-dependently suppress the protein and mRNA expressions of TLR2 and TLR4. TNF-alpha synthesis was also decreased upon TLR ligand stimulation in vitamin D(3)-treated monocytes. CONCLUSION: These results suggest that the inflammation triggered through TLR2 and TLR4 is important in the pathogenesis of BD. And it seems possible that vitamin D may be used as a therapeutic option by modulating TLR2 and TLR4 expression of monocytes in BD. PMID- 18411218 TI - Why do we choose rheumatology? Implications for future recruitment--results of the 2006 UK Trainee Survey. AB - OBJECTIVES: Against changes to junior doctor career structure under MMC (Modernizing Medical Careers), and uncertainty about the future place of rheumatology, we explored critical factors in choice of rheumatology as a speciality, and asked what factors might govern choices of prospective trainees. Using these data, we developed suggestions to enhance future recruitment. METHODS: A postal survey was sent to rheumatology specialist registrars (SpRs) on the Joint Committee for Higher Medical Training (JCHMT) database between December 2005 and January 2006, and concurrently by e-mail to the Rheumatologists at Training e-mail list. RESULTS: Seventy-three percent (165/227) of trainees responded. Of them, 89.1% had previous senior house officer (SHO) experience in rheumatology and 81.8% made a career decision in favour of rheumatology during their SHO post. The top four ranked factors influencing choice of rheumatology were SHO experience, subject matter, inspirational consultants and lifestyle aspects; 89.1% would still choose rheumatology now. Factors felt to be negatively influencing future trainees came under three key themes: poor student or postgraduate exposure, employment and service delivery issues (including concern over the future place of rheumatology in primary vs secondary care), and perceived poor profile of rheumatology. Factors positively influencing future candidates were subject matter, work/life balance and prior exposure to the speciality. CONCLUSIONS: Early postgraduate experience is key to choice of speciality. An overwhelming majority of trainees decide speciality during SHO experience. With ongoing changes in career structure, it is critical that rheumatology is incorporated into foundation and speciality training programmes and essential that continued measures are taken to improve the image of rheumatology. PMID- 18411219 TI - Course and prognosis of shoulder symptoms in general practice. AB - OBJECTIVES: To investigate the course and prognosis of shoulder pain in the first 6 months after presentation to the general practitioner. We separately studied patients with acute, subacute and chronic shoulder pain, as duration of symptoms at presentation has been shown to be the strongest predictor of outcome. METHODS: A prospective cohort study with 6 months follow-up was carried out in The Netherlands, including 587 patients with a new episode of shoulder pain. Patients were categorized as having acute (symptoms <6 weeks), subacute (6-12 weeks) or chronic (>3 months) shoulder pain. The course of shoulder pain, functional disability and quality of life was analysed over 6 months. Patient and disease characteristics, including physical and psychosocial factors, were investigated as possible predictors of outcome using multivariable regression analyses. RESULTS: Acute shoulder symptoms showed the most favourable course over 6 months follow-up, with larger pain reduction and improvement of functional disability. Patients with chronic shoulder symptoms showed the poorest results. The multivariable regression analysis showed that predictors of a better outcome at 6 months for acute shoulder pain were lower baseline disability scores and higher baseline pain intensity (explained variance 46%). Predictors of a better outcome for chronic shoulder pain were lower scores on pain catastrophizing and higher baseline pain intensity (explained variance 21%). CONCLUSIONS: The results indicate that, besides a different course of symptoms in patients presenting with acute or chronic shoulder pain, predictors of outcome may also differ with psychosocial factors being more important in chronic shoulder pain. PMID- 18411220 TI - Overuse and inappropriate prescribing of proton pump inhibitors in patients with Clostridium difficile-associated disease. AB - BACKGROUND: Clostridium difficile is the most common infectious cause of colitis and has been increasingly diagnosed in hospitalized patients. The number of prescriptions for proton pump inhibitors (PPIs) has also increased significantly over time. Few studies have reported an association between C. difficile associated disease (CDAD) and PPI use. AIM: To assess the extent and appropriateness of PPI prescribing in patients diagnosed with C. difficile infection. METHODS: We prospectively studied PPI prescriptions in 138 hospitalized patients diagnosed with C. difficile infection over a 4-month period. Clostridium difficile infections were diagnosed by the presence of C. difficile toxin in the stools. The appropriateness of prescriptions and relevant investigations were assessed by interview of patients and review of patient records. RESULTS: Sixty-four percent (88 of 138) of all patients who developed C. difficile infections were on PPIs. A valid indication for PPIs therapy was not apparent in 63% of the patients. CONCLUSION: There appears to be a widespread and inappropriate use of PPIs in hospital practice. Reduction of unnecessary PPIs use may be an additional strategy to reduce the incidence of this infection. PMID- 18411221 TI - Clinical and serological profile of primary biliary cirrhosis in young and elderly patients. PMID- 18411222 TI - Dynameomics: a multi-dimensional analysis-optimized database for dynamic protein data. AB - The Dynameomics project is our effort to characterize the native-state dynamics and folding/unfolding pathways of representatives of all known protein folds by way of molecular dynamics simulations, as described by Beck et al. (in Protein Eng. Des. Select., the first paper in this series). The data produced by these simulations are highly multidimensional in structure and multi-terabytes in size. Both of these features present significant challenges for storage, retrieval and analysis. For optimal data modeling and flexibility, we needed a platform that supported both multidimensional indices and hierarchical relationships between related types of data and that could be integrated within our data warehouse, as described in the accompanying paper directly preceding this one. For these reasons, we have chosen On-line Analytical Processing (OLAP), a multi-dimensional analysis optimized database, as an analytical platform for these data. OLAP is a mature technology in the financial sector, but it has not been used extensively for scientific analysis. Our project is further more unusual for its focus on the multidimensional and analytical capabilities of OLAP rather than its aggregation capacities. The dimensional data model and hierarchies are very flexible. The query language is concise for complex analysis and rapid data retrieval. OLAP shows great promise for the dynamic protein analysis for bioengineering and biomedical applications. In addition, OLAP may have similar potential for other scientific and engineering applications involving large and complex datasets. PMID- 18411223 TI - Dynameomics: design of a computational lab workflow and scientific data repository for protein simulations. AB - Dynameomics is a project to investigate and catalog the native-state dynamics and thermal unfolding pathways of representatives of all protein folds using solvated molecular dynamics simulations, as described in the preceding paper. Here we introduce the design of the molecular dynamics data warehouse, a scalable, reliable repository that houses simulation data that vastly simplifies management and access. In the succeeding paper, we describe the development of a complementary multidimensional database. A single protein unfolding or native state simulation can take weeks to months to complete, and produces gigabytes of coordinate and analysis data. Mining information from over 3000 completed simulations is complicated and time-consuming. Even the simplest queries involve writing intricate programs that must be built from low-level file system access primitives and include significant logic to correctly locate and parse data of interest. As a result, programs to answer questions that require data from hundreds of simulations are very difficult to write. Thus, organization and access to simulation data have been major obstacles to the discovery of new knowledge in the Dynameomics project. This repository is used internally and is the foundation of the Dynameomics portal site http://www.dynameomics.org. By organizing simulation data into a scalable, manageable and accessible form, we can begin to address substantial questions that move us closer to solving biomedical and bioengineering problems. PMID- 18411224 TI - Dynameomics: mass annotation of protein dynamics and unfolding in water by high throughput atomistic molecular dynamics simulations. AB - The goal of Dynameomics is to perform atomistic molecular dynamics (MD) simulations of representative proteins from all known folds in explicit water in their native state and along their thermal unfolding pathways. Here we present 188-fold representatives and their native state simulations and analyses. These 188 targets represent 67% of all the structures in the Protein Data Bank. The behavior of several specific targets is highlighted to illustrate general properties in the full dataset and to demonstrate the role of MD in understanding protein function and stability. As an example of what can be learned from mining the Dynameomics database, we identified a protein fold with heightened localized dynamics. In one member of this fold family, the motion affects the exposure of its phosphorylation site and acts as an entropy sink to offset another portion of the protein that is relatively immobile in order to present a consistent interface for protein docking. In another member of this family, a polymorphism in the highly mobile region leads to a host of disease phenotypes. We have constructed a web site to provide access to a novel hybrid relational/multidimensional database (described in the succeeding two papers) to view and interrogate simulations of the top 30 targets: http://www.dynameomics.org. The Dynameomics database, currently the largest collection of protein simulations and protein structures in the world, should also be useful for determining the rules governing protein folding and kinetic stability, which should aid in deciphering genomic information and for protein engineering and design. PMID- 18411225 TI - Site-directed mutagenesis of the hinge peptide from the hemagglutinin protein: enhancement of the pH-responsive conformational change. AB - Environmentally responsive proteins and peptides are increasingly finding utility in various engineered systems due to their ability to respond to the presentation of external stimuli. A classic example of this behavior is the influenza hemagglutinin (HA) fusion protein. At neutral pH, HA exists in a non-fusogenic state, but upon exposure to low pH, the conformation of the structure changes to expose a fusogenic peptide. During this structural change, massive rearrangements occur in a subunit of HA (HA2). Crystallography data has shown that a loop of 28 amino acids (residues 54-81) undergoes a dramatic transition from a random coil to an alpha-helix. This segment connects to two flanking helical regions (short and long) to form a long, continuous helix. Here, we report the results of site directed mutagenesis study on LOOP-36 to further understand the mechanism of this important stimulus-responsive peptide. The conformational transition of a bacterially expressed LOOP-36 was found to be less dramatic than has been previously reported. The systematic mutation of glutamate and histidine residues in the peptide to glutamines (glutamine scanning) did not impact the conformational behavior of the peptide, but the substitution of the glycine residue at position 22 with alanine resulted in significant pH-responsive behavior. Therefore this mutant stimulus-responsive peptide may be more valuable for future protein engineering and bionanotechnology efforts. PMID- 18411226 TI - Directed evolution on the cold adapted properties of TAB5 alkaline phosphatase. AB - Psychrophilic alkaline phosphatase (AP) from the Antarctic strain TAB5 was subjected to directed evolution in order to identify the key residues steering the enzyme's cold-adapted activity and stability. A round of random mutagenesis and further recombination yielded three thermostable and six thermolabile variants of the TAB5 AP. All of the isolated variants were characterised by their residual activity after heat treatment, Michaelis-Menten kinetics, activation energy and microcalorimetric parameters of unfolding. In addition, they were modelled into the structure of the TAB5 AP. Mutations which affected the cold adapted properties of the enzyme were all located close to the active site. The destabilised variants H135E and H135E/G149D had 2- and 3-fold higher kcat, respectively, than the wild-type enzyme. Wild-type AP has a complex heat-induced unfolding pattern while the mutated enzymes loose local unfolding transitions and have large shifts of the Tm values. Comparison of the wild-type and mutated TAB5 APs demonstrates that there is a delicate balance between the enzyme activity and stability and that it is possible to improve the activity and thermostability simultaneously as demonstrated in the case of the H135E/G149D variant compared to H135E. PMID- 18411227 TI - In vitro DNA recombination by L-Shuffling during ribosome display affinity maturation of an anti-Fas antibody increases the population of improved variants. AB - The use of random mutagenesis in concert with protein display technologies to rapidly select high affinity antibody variants is an established methodology. In some cases, DNA recombination has been included in the strategy to enable selection of mutations which act cooperatively to improve antibody function. In this study, the impact of L-Shuffling DNA recombination on the eventual outcome of an in vitro affinity maturation has been experimentally determined. Parallel evolution strategies, with and without a recombination step, were carried out and both methods improved the affinity of an anti-Fas single chain variable fragment (scFv). The recombination step resulted in an increased population of affinity improved variants. Moreover, the most improved variant, with a 22-fold affinity gain, emerged only from the recombination-based approach. An analysis of mutations preferentially selected in the recombined population demonstrated strong cooperative effects when tested in combination with other mutations but small, or even negative, effects on affinity when tested in isolation. These results underline the ability of combinatorial library approaches to explore very large regions of sequence space to find optimal solutions in antibody evolution studies. PMID- 18411229 TI - Role of inducible nitric oxide synthase in induction of RhoA expression in hearts from diabetic rats. AB - AIMS: Recent studies from our laboratory demonstrated that increased expression of the small GTP-binding protein RhoA and activation of the RhoA/rho kinase (ROCK) pathway play an important role in the contractile dysfunction associated with diabetic cardiomyopathy in hearts from streptozotocin (STZ)-induced diabetic rats. Nitric oxide (NO) has been reported to be a positive regulator of RhoA expression in vascular smooth muscle, and we have previously found that the expression of inducible NO synthase (iNOS) is increased in hearts from STZ diabetic rats. Therefore, in this study, we investigated the hypothesis that induction of iNOS positively regulates RhoA expression in diabetic rat hearts. METHODS AND RESULTS: To determine whether NO and iNOS could increase RhoA expression in the heart, cardiomyocytes from non-diabetic rats were cultured in the presence of the NO donor sodium nitroprusside (SNP) or lipopolysaccharide (LPS) in the absence and presence of the selective iNOS inhibitor, N(6)-(1 iminoethyl)-l-lysine dihydrochloride (L-NIL). In a second study, 1 week after induction of diabetes with STZ, rats were treated with L-NIL (3 mg/kg/day) for 8 more weeks to determine the effect of iNOS inhibition in vivo on RhoA expression and cardiac contractile function. Expression of iNOS was elevated in cardiomyocytes isolated from diabetic rat hearts. Both SNP and LPS increased RhoA expression in non-diabetic cardiomyocytes. The LPS-induced elevation in RhoA expression was accompanied by an increase in iNOS expression and prevented by L NIL. Treatment of diabetic rats with L-NIL led to a significant improvement in left ventricular developed pressure and rates of contraction and relaxation concomitant with normalization of total cardiac nitrite levels, RhoA expression, and phosphorylation of the ROCK targets LIM (Lin-11, Isl-1, Mec-3) kinase and ezrin/radixin/moesin. CONCLUSION: These data suggest that iNOS is involved in the increased expression of RhoA in diabetic hearts and that one of the mechanisms by which iNOS inhibition improves cardiac function is by preventing the upregulation of RhoA and its availability for activation. PMID- 18411228 TI - The molecular phenotype of human cardiac myosin associated with hypertrophic obstructive cardiomyopathy. AB - AIM: The aim of the study was to compare the functional and structural properties of the motor protein, myosin, and isolated myocyte contractility in heart muscle excised from hypertrophic cardiomyopathy patients by surgical myectomy with explanted failing heart and non-failing donor heart muscle. METHODS: Myosin was isolated and studied using an in vitro motility assay. The distribution of myosin light chain-1 isoforms was measured by two-dimensional electrophoresis. Myosin light chain-2 phosphorylation was measured by sodium dodecyl sulphate polyacrylamide gel electrophoresis using Pro-Q Diamond phosphoprotein stain. RESULTS: The fraction of actin filaments moving when powered by myectomy myosin was 21% less than with donor myosin (P = 0.006), whereas the sliding speed was not different (0.310 +/- 0.034 for myectomy myosin vs. 0.305 +/- 0.019 microm/s for donor myosin in six paired experiments). Failing heart myosin showed 18% reduced motility. One myectomy myosin sample produced a consistently higher sliding speed than donor heart myosin and was identified with a disease-causing heavy chain mutation (V606M). In myectomy myosin, the level of atrial light chain 1 relative to ventricular light chain-1 was 20 +/- 5% compared with 11 +/- 5% in donor heart myosin and the level of myosin light chain-2 phosphorylation was decreased by 30-45%. Isolated cardiomyocytes showed reduced contraction amplitude (1.61 +/- 0.25 vs. 3.58 +/- 0.40%) and reduced relaxation rates compared with donor myocytes (TT(50%) = 0.32 +/- 0.09 vs. 0.17 +/- 0.02 s). CONCLUSION: Contractility in myectomy samples resembles the hypocontractile phenotype found in end-stage failing heart muscle irrespective of the primary stimulus, and this phenotype is not a direct effect of the hypertrophy-inducing mutation. The presence of a myosin heavy chain mutation causing hypertrophic cardiomyopathy can be predicted from a simple functional assay. PMID- 18411230 TI - Selective attenuation of Toll-like receptor 2 signalling may explain the atheroprotective effect of sphingosine 1-phosphate. AB - AIMS: Vascular inflammation is a major atherogenic factor and Toll-like receptor (TLR) 2 ligands, including bacterial and serum lipoproteins, seem to be involved in atherogenesis. On this basis, we analysed the effect of lipoproteins and different lipid components on TLR2-dependent signalling. METHODS AND RESULTS: In TLR2-transfected human embryonic kidney 293 cells and human monocytes, oxidized low-density lipoproteins inhibited nuclear factor (NF)-kappaB-driven transcriptional activity and chemokine gene expression in response to TLR2 ligands. Sphingosine 1-phosphate (S1P) and oxidized palmitoyl-arachidonoyl phosphatidylcholine, but not lipoprotein-carried lysophospholipids, inhibited TLR2 activation. Silencing experiments in TLR2-transfected 293 cells showed that the S1P-mediated attenuation effect is mediated by S1P receptors type 1 and type 2. To address the physiological significance of these findings, additional experiments were performed in human peripheral blood monocytes and monocyte derived macrophages. In both cell types, S1P selectively attenuated TLR2 signalling, as NF-kappaB and extracellular signal-regulated kinase activation, but not c-Jun amino terminal kinase phosphorylation, were inhibited by physiologically relevant concentrations of S1P. Moreover, the attenuation of TLR2 signalling was partially reverted by pharmacological inhibition of phosphoinositide 3-kinase (PI3K) and Ras pathways. In addition, S1P inhibited the chemokine gene expression elicited by TLR2, but not by TLR4 ligands. CONCLUSION: These findings disclose a cross-talk mechanism between lipoprotein components and TLR in which engagement of S1P receptors exert selective attenuation of TLR2 dependent activation via PI3K and Ras signalling. A corollary to these data is that the negative cross-talk of S1P receptors and TLR2 signalling might be involved in the atheroprotective effects of S1P. PMID- 18411231 TI - Whole gene deletion of the hepatocyte nuclear factor-1beta gene in a patient with the prune-belly syndrome. PMID- 18411232 TI - Trichloroethylene disrupts cardiac gene expression and calcium homeostasis in rat myocytes. AB - We have been investigating the molecular mechanisms by which trichloroethylene (TCE) might induce cardiac malformations in the embryonic heart. Previous results indicated that TCE disrupted expression of genes encoding proteins involved in regulation of intracellular Ca2+, [Ca2+](i), in cardiac cells, including ryanodine receptor isoform 2 (Ryr2), and sarcoendoplasmatic reticulum Ca2+ ATPase, Serca2a. These observations are important in light of the notion that altered cardiac contractility can produce morphological defects. The hypothesis tested in this study is that the TCE-induced changes in gene expression of Ca2+ associated proteins resulted in altered Ca2+ flux regulation. We used real-time PCR and digital imaging microscopy to characterize effects of various doses of TCE on gene expression and Ca2+ response to vasopressin (VP) in rat cardiac H9c2 myocytes. We observed a reduction in Serca2a and Ryr2 expression at 12 and 48 h after exposure to TCE. In addition, we found significant differences in Ca2+ response to VP in cells treated with TCE doses as low as 10 parts per billion. Taken all together, our data strongly indicate that exposure to TCE disrupts the ability of myocytes to regulate cellular Ca2+ fluxes. Perturbation of calcium signaling alters cardiac cell physiology and signal transduction and may hint to morphogenetic consequences in the context of heart development. These results point to a novel area of TCE biology and, if confirmed in vivo, may help to explain the apparent cardio-specific toxicity of TCE exposure in the rodent embryo. PMID- 18411233 TI - A mixture of five phthalate esters inhibits fetal testicular testosterone production in the sprague-dawley rat in a cumulative, dose-additive manner. AB - Phthalate diesters are chemicals to which humans are ubiquitously exposed. Exposure to certain phthalates during sexual differentiation causes reproductive tract malformations in male rats. In the fetal rat, exposure to the phthalates benzylbutyl phthalate (BBP), di(n)butyl phthalate (DBP), and diethylhexyl phthalate (DEHP) decreases testicular testosterone production and insulin-like 3 hormone mRNA levels. We characterized the dose-response effects of six individual phthalates (BBP, DBP, DEHP, diethyl phthalate [DEP], diisobutyl phthalate [DiBP], and dipentyl phthalate [DPP]) on gestation day (GD) 18 testicular testosterone production following exposure of Sprague-Dawley rats on GD 8-18. BBP, DBP, DEHP, and DiBP were equipotent (ED50 of 440 +/- 16 mg/kg/day), DPP was about threefold more potent (ED50 = 130 mg/kg/day) and DEP had no effect on fetal testosterone production. We hypothesized that coadministration of these five antiandrogenic phthalates would reduce testosterone production in a dose-additive fashion because they act via a common mode of toxicity. In a second study, dams were dosed at 100, 80, 60, 40, 20, 10, 5, or 0% of the mixture. The top dose contained 1300 mg of total phthalates/kg/day including BBP, DBP, DEHP, DiBP (300 mg/kg/day per chemical), and DPP (100 mg DPP/kg/day). This mixture ratio was selected such that each phthalate would contribute equally to the reduction in testosterone. As hypothesized, testosterone production was reduced in a dose-additive manner. Several of the individual phthalates and the mixture also induced fetal mortality, due to pregnancy loss. These data demonstrate that individual phthalates with a similar mechanism of action can elicit cumulative, dose additive effects on fetal testosterone production and pregnancy when administered as a mixture. PMID- 18411234 TI - Dose- and route-dependent teratogenicity, toxicity, and pharmacokinetic profiles of the hedgehog signaling antagonist cyclopamine in the mouse. AB - The Hedgehog (Hh) signaling pathway is an essential regulator of embryonic development and appears to play important roles in postnatal repair and cancer progression and metastasis. The teratogenic Veratrum alkaloid cyclopamine is a potent Hh antagonist and is used experimentally both in vitro and in vivo to investigate the role of Hh signaling in diverse biological processes. Here, we set out to establish an administration regimen for cyclopamine-induced teratogenicity in the mouse. The dysmorphogenic concentration of cyclopamine was determined in vitro via mouse whole-embryo culture assays to be 2.0 microM. We administered cyclopamine to female C57BL/6J mice at varied doses by oral gavage, ip injection, or osmotic pump infusion and assessed toxicity and pharmacokinetic (PK) models. Bolus administration was limited by toxicity and rapid clearance. In vivo cyclopamine infusion at 160 mg/kg/day yielded a dam serum steady-state concentration of approximately 2 microM with a corresponding amniotic fluid concentration of approximately 1.5 microM. Gross facial defects were induced in 30% of cyclopamine-exposed litters, with affected embryos exhibiting cleft lip and palate. This is the first report describing the PKs and teratogenic potential of cyclopamine in the mouse and demonstrates that transient Hh signaling inhibition induces facial clefting anomalies in the mouse that mimic common human birth defects. PMID- 18411235 TI - Nasal continuous positive airway pressure with heliox versus air oxygen in infants with acute bronchiolitis: a crossover study. AB - OBJECTIVE: The purpose of this work was to evaluate the effects of administering either heliox or air oxygen in combination with nasal continuous positive airway pressure in infants with refractory bronchiolitis. PATIENT AND METHODS: We conducted a prospective, interventional, single-center, crossover study in a teaching hospital including infants 1 month to 2 years of age, consecutively admitted to the PICU from February 2004 to February 2005 for treatment of severe acute bronchiolitis unresponsive to therapy. Patients with a clinical score (Modified Wood's Clinical Asthma Score) of >5, arterial oxygen saturation of <92%, or transcutaneous CO(2) pressure of >50 mmHg despite supportive therapy, nebulized L-epinephrine, and heliox therapy through a nonrebreathing reservoir face mask were eligible. During the study period, 40 infants with bronchiolitis were admitted to the PICU; 12 fulfilled inclusion criteria. A predetermined balanced sequential allocation to either 30 minutes of treatment with nasal continuous positive airway pressure with heliox or to air-oxygen nasal continuous positive airway pressure was performed. Measurements were taken at baseline and after 30 minutes of each treatment. RESULTS: Baseline mean values were as follows: nasal continuous positive airway pressure of 7.2 cmH(2)O; clinical score of 7.7 points; transcutaneous CO(2) pressure of 61.6 mmHg; and arterial oxygen saturation of 88.6%, with the fraction of inspired oxygen at 35.4%. Clinical score, transcutaneous CO(2) pressure, and arterial oxygen saturation improved during the study time with both heliox-nasal continuous positive airway pressure and air-oxygen-nasal continuous positive airway pressure: after 1 hour, the clinical score fell 1.7 points, transcutaneous CO(2) pressure decreased 8.2 mmHg, and arterial oxygen saturation increased by 7.7%. Improvement in clinical score was double with heliox-nasal continuous positive airway pressure compared with the air-oxygen-nasal continuous positive airway pressure (2.12 vs 1.08 points), and the fall in the transcutaneous CO(2) pressure was greater with heliox-nasal continuous positive airway pressure compared with air-oxygen-nasal continuous positive airway pressure (9.7 vs 5.4 mm Hg). There was no difference in arterial oxygen saturation between groups. No patients required endotracheal intubation. No adverse effects attributable to either of the study interventions were detected. CONCLUSIONS: Nasal continuous positive airway pressure improves the clinical score and the CO(2) elimination of infants with refractory bronchiolitis. These positive effects are significantly enhanced when nasal continuous positive airway pressure is combined with heliox instead of air oxygen. Both techniques are noninvasive, seem safe, and may reduce the need for endotracheal intubation. PMID- 18411236 TI - Evaluation of long-term treatment of children with congenital lactic acidosis with dichloroacetate. AB - OBJECTIVE: The purpose of this research was to report results on long-term administration of dichloroacetate in 36 children with congenital lactic acidosis who participated previously in a controlled trial of this drug. PATIENTS AND METHODS: We conducted a randomized control trial, followed by an open-label study. Data were analyzed for each patient from the time they began treatment through May 2005. RESULTS: Subject exposure to dichloroacetate totaled 110.42 years. Median height and weight increased over time, but the standardized values declined slightly and remained below the first percentile. There were no significant changes in biochemical metabolic indices, except for a 2% rise in total protein and a 22% increase in 24-hour urinary oxalate. Both the basal and carbohydrate meal-induced rises in lactate were blunted by dichloroacetate. The median cerebrospinal fluid lactate also decreased over time. Conduction velocity decreased and distal latency increased in peroneal nerves. Mean 3-year survival for all of the subjects was 79%. CONCLUSIONS: Oral dichloroacetate is generally well tolerated in young children with congenital lactic acidosis. Although continued dichloroacetate exposure is associated with evidence of peripheral neuropathy, it cannot be determined whether this is attributable mainly to the drug or to progression of underlying disease. PMID- 18411238 TI - Perceived anxiety and plasma cortisol concentrations following rock climbing with differing safety rope protocols. AB - OBJECTIVES: To examine how different safety rope protocols impact on subjective anxiety and self-confidence levels and plasma cortisol concentrations and the relationship between subjective states and cortisol during rock climbing. METHODS: Participants (n = 12) were tested in three climbing conditions that were designed to invoke low, moderate and high physical and mental stress. Plasma cortisol concentrations were collected before and after climbing and participants reported subjective anxiety and self-confidence states for each climb. RESULTS: Repeated measures analysis of variance showed significant differences between conditions for somatic anxiety (F(2, 22) = 7.74, p = 0.009), self-confidence (F(2, 22) = 9.52, p = 0.001) and change in plasma cortisol concentration (F(2, 22) = 3.71, p = 0.041). Preplanned polynomial comparisons showed that these were linear effects; somatic anxiety was higher in the higher stress conditions whilst self-confidence was lower. Plasma cortisol concentration change was also linear. Regression analyses showed cubic relationships between plasma cortisol concentration and cognitive anxiety (R(2) = 0.452), self-confidence (R(2) = 0.281) and somatic anxiety (R(2) = 0.268). CONCLUSIONS: There is a relationship between plasma cortisol concentration and subjective anxiety and self-confidence states during rock climbing. Changes in the way the safety rope is organised can impact on anxiety, cortisol concentration and self-confidence during rock climbing. PMID- 18411237 TI - Effect of low-dose naloxone infusion on fentanyl requirements in critically ill children. AB - OBJECTIVE: Sedating critically ill patients often involves prolonged opioid infusions causing opioid tolerance. Naloxone has been hypothesized to limit opioid tolerance by decreasing adenylate cyclase/cyclic adenosine monophosphate activation. The study purpose was to investigate the effect of low-dose naloxone on the maximum cumulative daily fentanyl dose in critically ill children. METHODS: We conducted a double-blinded, randomized, placebo-control trial from December 2002 through July 2004 in a university PICU. We enrolled 82 children age 1 day to 18 years requiring mechanical ventilation and fentanyl infusions anticipated to last for >4 days were eligible for enrollment. Those receiving additional oral analgesia or sedation, having a history of drug dependence or withdrawal, or having significant neurologic, renal, or hepatic disease were excluded. In addition to fentanyl infusions, patients received low-dose naloxone or placebo infusions. Medications were adjusted using the Modified Motor Activity Assessment Scale. Withdrawal was monitored using the Modified Narcotic Withdrawal Scale. Intervention was a low-dose naloxone infusion (0.25 microg/kg per hour) and the main outcome variable was the maximum cumulative daily fentanyl dose (micrograms per kilogram per day). RESULTS: There was no difference in the maximum cumulative daily fentanyl dose between patients treated with naloxone (N = 37) or those receiving placebo (N = 35). Adjustment for the starting fentanyl dose also failed to reveal group differences. Total fentanyl dose received throughout the study in the naloxone group (360 microg/kg) versus placebo (223 microg/kg) was not statistically different. Placebo patients trended toward fewer rescue midazolam boluses (10.7 vs 17.8), lower total midazolam dose (11.6 mg/kg vs 23.9 mg/kg), and fewer rescue fentanyl boluses (18.5 vs 23.9). CONCLUSIONS: We conclude that administration of low-dose naloxone (0.25 microg/kg per hour) does not decrease fentanyl requirements in critically ill, mechanically ventilated children. PMID- 18411239 TI - Randomized efficacy trial of early preconception counseling for diabetic teens (READY-girls). AB - OBJECTIVE: To develop and assess the feasibility of an early preconception counseling program for adolescents called READY-Girls (Reproductive-health Education and Awareness of Diabetes in Youth for Girls). RESEARCH DESIGN AND METHODS: A total of 53 adolescent females with type 1 diabetes between 16 and 19.9 years of age were randomized into groups receiving a CD-ROM, a book, or standard care (control) and given one comprehensive session. Outcomes were assessed at baseline, immediately after, and at 3 months. RESULTS: Teens who received the CD and those who received the book demonstrated significant (P < or = 0.05) sustained improvement (over 3 months) in knowledge, perceived benefits of both receiving preconception counseling and using effective family planning, and perceived more support with reproductive health issues. CONCLUSIONS: Clinical feasibility of the program was demonstrated. Both the CD and the book appeared to be efficacious formats for the short term. Future studies should examine repeated boosters of a CD and a book, which are not meant to replace but rather to reinforce and supplement health professional education. PMID- 18411240 TI - Long-term follow-up of oral glucose tolerance test-derived glucose tolerance and insulin secretion and insulin sensitivity indexes in subjects with glucokinase mutations (MODY2). AB - OBJECTIVE: We investigated the natural history of glucokinase (GCK)-related maturity-onset diabetes of the young type 2 (MODY2), notably the factors associated with deterioration of hyperglycemia over time. RESEARCH DESIGN AND METHODS: We report an 11-year follow-up of glucose tolerance and indexes of insulin secretion and insulin sensitivity derived from oral glucose tolerance tests in 33 MODY2 subjects. RESULTS: The variation between tests of glucose tolerance (expressed as the area under the glucose curve) was 6.9 +/- 3.2% (mean +/- SEM), but individual results ranged from -20 to 61%. Deterioration of glucose tolerance between tests was associated with decreased insulin sensitivity, while insulin secretion remained stable. CONCLUSIONS: Glucose tolerance can remain stable over many years in subjects with MODY2 due to the relative stability of the GCK-related beta-cell defect. However, the development of insulin resistance may have an important role in the deterioration of the glucose tolerance and in the long-term evolution of the disorder. PMID- 18411241 TI - Longitudinal association of glucose metabolism with retinopathy: results from the Australian Diabetes Obesity and Lifestyle (AusDiab) study. AB - OBJECTIVE: We determined the longitudinal association of glucose metabolism with retinopathy in a sample of the Australian population. RESEARCH DESIGN AND METHODS: The Australian Diabetes Obesity and Lifestyle (AusDiab) study is a national, longitudinal study of adults aged > or =25 years from 42 randomly selected areas of Australia. Retinopathy was assessed at baseline in 1999-2000 and 5 years later in 2004-2005 in participants identified as having diabetes (based on self-report and oral glucose tolerance test) and impaired glucose metabolism and in a random sample with normal glucose tolerance. Complete retinal data were available for 1,192 participants. Photographs were graded at two time points according to a simplified version of the Wisconsin grading system. RESULTS: The 5-year incidences of retinopathy were 13.9 and 3.0% among those with known and newly diagnosed diabetes at baseline, respectively. Of those who developed incident newly diagnosed diabetes at follow-up, 11.9% had retinopathy at baseline compared with 5.6% of those who did not progress to incident newly diagnosed diabetes (P = 0.037). After adjustment for factors identified as risk factors for diabetes, individuals with retinopathy signs at baseline were twice as likely to develop incident newly diagnosed diabetes compared with those who did not have retinopathy signs at baseline. CONCLUSIONS: The 5-year incidence of retinopathy was 13.9% among individuals with known diabetes. Nondiabetic individuals with retinopathy signs at baseline had a twofold higher risk of developing incident newly diagnosed diabetes 5 years later. This result provides further evidence that mild retinopathy signs may be a preclinical marker of underlying microvascular disease and future diabetes risk. PMID- 18411243 TI - Clinical characteristics of carbohydrate-deficient transferrin (%disialotransferrin) measured by HPLC: sensitivity, specificity, gender effects, and relationship with other alcohol biomarkers. AB - AIMS: The sensitivity and specificity of the relative disialotransferrin amount (%DST), considered the primary single target for measurement of the alcohol biomarker carbohydrate-deficient transferrin (CDT), were compared with the absolute CDT amount determined by the CDTect assay and with GGT and AST. METHODS: Serum samples (n = 1387) were collected within the WHO/ISBRA Study on State and Trait Markers of Alcohol Use and Dependence. The subjects had been classified as "nondrinkers" (26%), "light/moderate drinkers" (50%), or "heavy drinkers" (24%) by use of the WHO/ISBRA Interview Schedule. An HPLC candidate reference method for CDT was used to quantify individual transferrin glycoforms. RESULTS: No gender difference in %DST was noted for nondrinkers, but light/moderate and heavy drinking males had significantly higher levels than females. Of the alcohol biomarkers examined, %DST showed the strongest correlation with self-reported alcohol intake, except for female heavy drinkers. The area under the %DST ROC curve for male (0.83) and female (0.82) heavy drinkers was significantly higher compared with CDT by CDTect (0.68) and GGT (0.69). At the 40, 60, or 80 g ethanol/day thresholds, %DST showed lower test sensitivity in women but there was no significant gender difference in overall accuracy according to ROC curve analysis. CONCLUSIONS: %DST measured by HPLC showed overall higher sensitivity for "heavy drinking" and better correlation with recent high alcohol intake, compared with the absolute CDT amount, and GGT and AST. The observation that several "light/moderate drinkers" had elevated %DST levels and some also a measurable asialotransferrin indicated misclassification with the WHO/ISBRA Interview Schedule and emphasize the limitations of self-reports of drinking. PMID- 18411244 TI - SNX18 is an SNX9 paralog that acts as a membrane tubulator in AP-1-positive endosomal trafficking. AB - SNX9, SNX18 and SNX30 constitute a separate subfamily of PX-BAR-containing sorting nexin (SNX) proteins. We show here that most tissues express all three paralogs, and immunoprecipitation and immunofluorescence experiments demonstrated that the SNX9-family proteins act as individual entities in cells. Their SH3 domains displayed a high selectivity for dynamin 2, and the PX-BAR units had the capacity to tubulate membranes when expressed in HeLa cells. As previously described for the PX-BAR domain of SNX9 (SNX9-PX-BAR), purified SNX18-PX-BAR caused liposome tubulation in vitro and had a binding preference for PtdIns(4,5)P(2). However, contrary to SNX9, which primarily acts in clathrin mediated endocytosis at the plasma membrane, endogenous SNX18 localized to AP-1- and PACS1-positive endosomal structures, which were devoid of clathrin and resistant to Brefeldin A. Moreover, a gamma-adaptin recognition motif was defined in a low-complexity region of SNX18, and a complex of endogenous SNX18 and AP-1 could be immunoprecipitated after Brefeldin A treatment. Overexpression of SNX18 sequestered AP-1 from peripheral endosomes and resulted in the formation of short SNX18-decorated tubes with distinct dynamin puncta. The results indicate that SNX9-family members make up discrete membrane-scission units together with dynamin, and suggest that SNX18 mediates budding of carriers for AP-1-positive endosomal trafficking. PMID- 18411245 TI - A new role for kinesin-directed transport of Bik1p (CLIP-170) in Saccharomyces cerevisiae. AB - Bik1p is the budding yeast counterpart of the CLIP-170 family of microtubule plus end tracking proteins, which are required for dynein localization at plus ends and dynein-dependent spindle positioning. CLIP-170 proteins make up a CAP-Gly microtubule-binding domain, which sustains their microtubule plus-end tracking behaviour. However, in yeast, Bik1p travels towards plus ends as a cargo of the plus-end-directed kinesin Kip2p. Additionally, Kip2p behaves as a plus-end tracking protein; hence, it has been proposed that Bik1p might track plus ends principally as a cargo of Kip2p. Here, we examined Bik1p localization in yeast strains expressing mutant tubulin lacking the C-terminal amino acid (Glu tubulin; lacking Phe), the interaction of which with Bik1p is severely impaired compared with wild type. In Glu-tubulin strains, despite the presence of robust Kip2p comets at microtubule plus ends, Bik1p failed to track plus ends. Despite Bik1p depletion at plus ends, dynein positioning at the same plus ends was unperturbed. Video microscopy and genetic evidence indicated that dynein was transported at plus ends in a Kip2p-Bik1p-dependent manner, and was then capable of tracking Bik1p-depleted plus ends. These results indicate that Bik1p interactions with tubulin are important for Bik1p plus-end tracking, and suggest alternative pathways for Bik1p-Kip2p-dependent dynein localization at plus ends. PMID- 18411246 TI - Mug27 is a meiosis-specific protein kinase that functions in fission yeast meiosis II and sporulation. AB - Several meiosis-specific proteins of Schizosaccharomyces pombe play essential roles in meiotic progression. We report here that a novel meiosis-specific protein kinase, Mug27 (also known as Ppk35), is required for proper spore formation. This kinase is expressed by the mug27(+) gene, which is abruptly transcribed after horsetail movement. This transcription is maintained until the second meiotic division. Green fluorescent protein (GFP)-tagged Mug27 appears at the start of prometaphase I, localizes to the spindle pole body (SPB) and then translocates to the forespore membrane (FSM) at late anaphase II. In the mug27Delta strain, smaller spores are produced compared with those of the mug27(+) strain. Moreover, spore viability was reduced by half or more compared with that of the mug27(+) strain. The protein-kinase activity of Mug27 appears to be important for its function: the putative kinase-dead Mug27 mutant had similar phenotypes to mug27Delta. Our results here indicate that the Mug27 kinase localizes at the SPB and regulates FSM formation and sporulation. PMID- 18411247 TI - Pathway selection to the axon depends on multiple targeting signals in NgCAM. AB - Similar to most differentiated cells, both neurons and epithelial cells elaborate distinct plasma membrane domains that contain different membrane proteins. We have previously shown that the axonal cell-adhesion molecule L1/NgCAM accumulates on the axonal surface by an indirect transcytotic pathway via somatodendritic endosomes. MDCK epithelial cells similarly traffic NgCAM to the apical surface by transcytosis. In this study, we map the signals in NgCAM required for routing via the multi-step transcytotic pathway. We identify both a previously mapped tyrosine-based signal as a sufficient somatodendritic targeting signal, as well as a novel axonal targeting signal in the cytoplasmic tail of NgCAM. The axonal signal is glycine and serine rich, but only the glycine residues are required for activity. The somatodendritic signal is cis-dominant and needs to be inactivated in order for the axonal signal to be executed. Additionally, we show that the axonal cytoplasmic signal promotes apical targeting in MDCK cells. Transcytosis of NgCAM to the axon thus requires the sequential regulated execution of multiple targeting signals. PMID- 18411248 TI - Organellar dynamics during the cell cycle of Toxoplasma gondii. AB - The protozoan phylum Apicomplexa encompasses approximately 5000 species of obligate intracellular parasites, including those responsible for malaria and toxoplasmosis. Rather than dividing by binary fission, apicomplexans use a remarkable mechanism for replication, assembling daughters de novo within the cytoplasm. Here, we exploit time-lapse microscopy of fluorescent markers targeted to various subcellular structures in Toxoplasma gondii tachyzoites to determine how these unicellular eukaryotes efficiently package a complete set of organelles, maintaining the highly polarized organization necessary for host cell invasion and pathogenesis. Golgi division and elongation of the apicoplast are among the first morphologically observable events, associated with an unusual pattern of centriolar migration. Daughter parasites are assembled on cytoskeletal scaffolding, whose growth proceeds from the apical end, first encapsulating the divided Golgi. Further extension of the cytoskeletal scaffold results in partitioning of the apicoplast, nucleus, endoplasmic reticulum, and finally the mitochondrion, which enters the developing daughters rapidly, but only very late during the division cycle. The specialized secretory organelles (micronemes and rhoptries) form de novo. This distinctive pattern of replication -- in which organellar segregation spans approximately 75% of the cell cycle, completely encompassing S phase -- suggests an unusual mechanism of cell cycle regulation. PMID- 18411249 TI - Binding of ATP to UAP56 is necessary for mRNA export. AB - The major-histocompatibility-complex protein UAP56 (BAT1) is a DEAD-box helicase that is deposited on mRNA during splicing. UAP56 is retained on spliced mRNA in an exon junction complex (EJC) or, alternatively, with the TREX complex at the 5' end, where it might facilitate the export of the spliced mRNA to the cytoplasm. Using confocal microscopy, UAP56 was found to be concentrated in RNA-splicing speckled domains of nuclei but was also enriched in adjacent nuclear regions, sites at which most mRNA transcription and splicing occur. At speckled domains, UAP56 was in complexes with the RNA-splicing and -export protein SRm160, and, as measured by FRAP, was in a dynamic binding equilibrium. The application of an in vitro FRAP assay, in which fluorescent nuclear proteins are photobleached in digitonin-extracted cells, revealed that the equilibrium binding of UAP56 in complexes at speckled domains was directly regulated by ATP binding. This was confirmed using a point mutant of UAP56 that did not bind ATP. Point mutation of UAP56 to eliminate ATP binding did not affect RNA splicing, but strongly inhibited the export of mRNA to the cytoplasm. PMID- 18411250 TI - Dissecting the role of PtdIns(4,5)P2 in endocytosis and recycling of the transferrin receptor. AB - Endocytosis and recycling of membrane proteins are key processes for nutrient uptake, receptor signaling and synaptic transmission. Different steps in these fission and fusion cycles have been proposed to be regulated by physiological changes in plasma membrane (PM) phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P(2)] concentration. Here, we use a chemical enzyme-translocation strategy to rapidly reduce PM PtdIns(4,5)P(2) levels while monitoring clathrin mediated endocytosis and recycling. PtdIns(4,5)P(2) hydrolysis blocked transferrin receptor endocytosis and led to a marked increase in the concentration of transferrin receptors in the PM, suggesting that endocytosis is more sensitive to changes in PtdIns(4,5)P(2) than recycling. Reduction of PM PtdIns(4,5)P(2) levels led to a near complete dissociation of Adaptor protein 2 (AP-2) from the PM but had only a small effect on clathrin assembly. This argues that receptor-mediated PtdIns(4,5)P(2) reduction preferentially suppresses AP-2 mediated targeting of cargo to endocytic sites rather than the assembly of clathrin coats or recycling of endocytic vesicles. PMID- 18411251 TI - Downregulation by lipopolysaccharide of Notch signaling, via nitric oxide. AB - The Notch signaling pathway appears to perform an important function in inflammation. Here, we present evidence to suggest that lipopolysaccharide (LPS) suppresses Notch signaling via the direct modification of Notch by the nitration of tyrosine residues in macrophages. In the RAW264.7 macrophage cell line and in rat primary alveolar macrophages, LPS was found to inhibit Notch1 intracellular domain (Notch1-IC) transcription activity, which could then be rescued by treatment with N(G)-nitro-l-arginine, a nitric oxide synthase (NOS) inhibitor. Nitric oxide (NO), which was produced in cells that stably express endothelial NOS (eNOS) and brain NOS (bNOS), also induced the inhibition of Notch1 signaling. The NO-induced inhibition of Notch1 signaling remained unchanged after treatment with 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ), a guanylyl-cyclase inhibitor, and was not found to be mimicked by 8-bromo-cyclic GMP in the primary alveolar macrophages. With regards to the control of Notch signaling, NO appears to have a significant negative influence, via the nitration of Notch1-IC, on the binding that occurs between Notch1-IC and RBP-Jk, both in vitro and in vivo. By intrinsic fluorescence, we also determined that nitration could mediate conformational changes of Notch1-IC. The substitution of phenylalanine for tyrosine at residue 1905 in Notch1-IC abolished the nitration of Notch1-IC by LPS. Overall, our data suggest that an important relationship exists between LPS mediated inflammation and the Notch1 signaling pathway, and that this relationship intimately involves the nitration of Notch1-IC tyrosine residues. PMID- 18411252 TI - Dynamic analysis identifies novel roles for DLG-1 subdomains in AJM-1 recruitment and LET-413-dependent apical focusing. AB - Cell-cell junctions are composed of a diverse array of specialized proteins that are necessary for the movement and integrity of epithelia. Scaffolding molecules, such as membrane-associated guanylate kinases (MAGUKs) contain multiple protein protein interaction domains that integrate these proteins into macromolecular complexes at junctions. We have used structure-function experiments to dissect the role of domains of the Caenorhabditis elegans MAGUK DLG-1, a homolog of Drosophila Discs large and vertebrate SAP97. DLG-1 deletion constructs were analyzed in directed yeast two-hybrid tests as well as in vivo in a dlg-1 null mutant background. Our studies identify novel roles for several key domains. First, the L27 domain of DLG-1 mediates the physical interaction of DLG-1 with its binding partner, AJM-1, as well as DLG-1 multimerization. Second, the PDZ domains of DLG-1 mediate its association with the junction. Third, using dynamic in vivo imaging, we demonstrate that the SH3 domain is required for rapid lateral distribution of DLG-1 via a LET-413/Scribble-dependent pathway. Finally, we found that inclusion of the SH3 domain can ameliorate dlg-1 mutant phenotypes, but full rescue of lethality required the complete C terminus, which includes the GUK and Hook domains, thereby demonstrating the importance of the C-terminus for DLG-1 function. Our results represent the first in vivo analysis of requirements for the L27 domain of a Discs-large/SAP97 protein, identify a crucial LET 413/Scribble regulatory motif and provide insight into how MAGUK subdomains function to maintain epithelial integrity during development. PMID- 18411253 TI - TbG63, a golgin involved in Golgi architecture in Trypanosoma brucei. AB - Golgins are coiled-coil proteins that have been implicated in the structure and function of the Golgi complex. Here, we identify and characterize a trypanosomal golgin, TbG63, showing that it has a C-terminal membrane anchor and an N-terminus that projects into the cytoplasm. TbG63 in procyclic parasites is localized to the Golgi and interacts with the active, GTP-form of TbRab1A. Overexpression of TbG63 has dramatic effects on Golgi architecture -- effects that require the N terminus -- whereas depletion has little, if any, effect on the growth rate. By contrast, in the bloodstream form of the parasite, depletion of TbG63 slows growth, although it has no obvious effect on the transport of a variant surface glycoprotein (VSG) or on Golgi structure. TbG63 might be a useful tool to study the structure and functioning of the Golgi complex. PMID- 18411254 TI - The molecular mechanism underlying Roberts syndrome involves loss of ESCO2 acetyltransferase activity. AB - Roberts syndrome/SC phocomelia (RBS) is an autosomal recessive disorder with growth retardation, craniofacial abnormalities and limb reduction. Cellular alterations in RBS include lack of cohesion at the heterochromatic regions around centromeres and the long arm of the Y chromosome, reduced growth capacity, and hypersensitivity to DNA damaging agents. RBS is caused by mutations in ESCO2, which encodes a protein belonging to the highly conserved Eco1/Ctf7 family of acetyltransferases that is involved in regulating sister chromatid cohesion. We identified 10 new mutations expanding the number to 26 known ESCO2 mutations. We observed that these mutations result in complete or partial loss of the acetyltransferase domain except for the only missense mutation that occurs in this domain (c.1615T>G, W539G). To investigate the mechanism underlying RBS, we analyzed ESCO2 mutations for their effect on enzymatic activity and cellular phenotype. We found that ESCO2 W539G results in loss of autoacetyltransferase activity. The cellular phenotype produced by this mutation causes cohesion defects, proliferation capacity reduction and mitomycin C sensitivity equivalent to those produced by frameshift and nonsense mutations associated with decreased levels of mRNA and absence of protein. We found decreased proliferation capacity in RBS cell lines associated with cell death, but not with increased cell cycle duration, which could be a factor in the development of phocomelia and cleft palate in RBS. In summary, we provide the first evidence that loss of acetyltransferase activity contributes to the pathogenesis of RBS, underscoring the essential role of the enzymatic activity of the Eco1p family of proteins. PMID- 18411255 TI - The S18Y polymorphic variant of UCH-L1 confers an antioxidant function to neuronal cells. AB - A number of studies have associated the S18Y polymorphic variant of UCH-L1 with protection from sporadic Parkinson's Disease (PD). The mechanism involved in this protective function is unknown, but has generally been assumed to be linked to the ubiquitin-proteasome system (UPS). In the current study, we have investigated the effects of overexpression of UCH-L1 and its variants, including S18Y, in neuronal cells. We find that S18Y, but not WT, UCH-L1 confers a specific antioxidant protective function when expressed at physiological levels in human neuroblastoma cells and primary cortical neurons. In contrast, neither WT nor S18Y UCH-L1 appear to directly impact the proteasome, although they both lead to stabilization of free ubiquitin. Lack of WT mouse UCH-L1 in neurons derived from gad mice led to a decrease of free ubiquitin, but no overall decrease in UPS function or enhanced sensitivity to oxidative stress. We conclude that the S18Y variant of UCH-L1 confers a novel antioxidant function that is not present in the WT form and that this function may underlie the protective effects of this variant in certain PD populations. Our results furthermore provide indirect evidence for the importance of oxidative stress as a pathogenetic factor in certain forms of sporadic PD. PMID- 18411257 TI - Plant genome horizons: Michael Bennett's contribution to genome research. PMID- 18411258 TI - Retroelement insertional polymorphisms, diversity and phylogeography within diploid, D-genome Aegilops tauschii (Triticeae, Poaceae) sub-taxa in Iran. AB - BACKGROUND AND AIMS: The diploid goat grass Aegilops tauschii (2n = 2x = 14) is native to the Middle East and is the D-genome donor to hexaploid bread wheat. The aim of this study was to measure the diversity of different subspecies and varieties of wild Ae. tauschii collected across the major areas where it grows in Iran and to examine patterns of diversity related to the taxa and geography. METHODS: Inter-retroelement amplified polymorphism (IRAP) markers were used to analyse the biodiversity of DNA from 57 accessions of Ae. tauschii from northern and central Iran, and two hexaploid wheats. Key Results Eight IRAP primer combinations amplified a total of 171 distinct DNA fragments between 180 and 3200 bp long from the accessions, of which 169 were polymorphic. On average, about eight fragments were amplified with each primer combination, with more bands being amplified from accessions from the north-west of the country than from other accessions. CONCLUSIONS: The IRAP markers showed high levels of genetic diversity. Analysis of all accessions together did not allow the allocation of individuals to taxa based on morphology, but showed a tendency to put accessions from the north-west apart from others regions. It is speculated that this could be due to different activity of retroelements in the different regions. Within the two taxa with most accessions, there was a range of IRAP genotypes that could be correlated closely with geographical origin. This supports suggestions that the centre of origin of the species is towards the south-east of the Caspian Sea. IRAP is an appropriate marker system to evaluate genetic diversity and evolutionary relationships within the taxa, but it is too variable to define the taxa themselves, where more slowly evolving morphological, DNA sequence or chromosomal makers may be more appropriate. PMID- 18411260 TI - Aggressiveness of cancer-care near the end-of-life in Korea. AB - OBJECTIVE: The aim of this study was to examine the appropriateness of chemotherapy and care in Korean cancer patients near the end-of-life. METHODS: We designed a retrospective cohort composed of patients diagnosed as having metastatic cancer and who received palliative chemotherapy at Seoul National University Hospital in 2002. Two hundred and ninety-eight patients who died of cancer were evaluated in terms of the appropriateness of the cancer-care they received, including chemotherapy. RESULTS: Median duration of chemotherapy was 6.02 months compared with 8.67 months for median overall survival. The median period between last chemotherapy and death was 2.02 months. Of the 298 patients, 50.3% received chemotherapy during the last 2 months of life. Furthermore, 17 patients (5.7%) died within 2 weeks after receiving chemotherapy. The proportion who visited an emergency room (ER) more than once during the last months of life was 33.6%, and the average number of ER visits after a diagnosis of cancer was 1.72. Only 9.1% of patients were referred to a hospice consultation service and only 11.7% of patients agreed with written DNR. CONCLUSIONS: Among patients who died of cancer, significant proportions were found to have received chemotherapy up to the end-of-life and to have visited ERs. Hospice referrals and discussions about DNR were not conducted well during the end-of-life period in Korea. PMID- 18411261 TI - Entropy-driven cAMP-dependent allosteric control of inhibitory interactions in exchange proteins directly activated by cAMP. AB - Exchange proteins directly activated by cAMP (EPACs) are guanine nucleotide exchange factors for the small GTPases Rap1 and Rap2 and represent a key receptor for the ubiquitous cAMP second messenger in eukaryotes. The cAMP-dependent activation of apoEPAC is typically rationalized in terms of a preexisting equilibrium between inactive and active states. Structural and mutagenesis analyses have shown that one of the critical determinants of the EPAC activation equilibrium is a cluster of salt bridges formed between the catalytic core and helices alpha1 and alpha2 at the N terminus of the cAMP binding domain and commonly referred to as ionic latch (IL). The IL stabilizes the inactive states in a closed topology in which access to the catalytic domain is sterically occluded by the regulatory moiety. However, it is currently not fully understood how the IL is allosterically controlled by cAMP. Chemical shift mapping studies consistently indicate that cAMP does not significantly perturb the structure of the IL spanning sites within the regulatory region, pointing to cAMP-dependent dynamic modulations as a key allosteric carrier of the cAMP-signal to the IL sites. Here, we have therefore investigated the dynamic profiles of the EPAC1 cAMP binding domain in its apo, cAMP-bound, and Rp-cAMPS phosphorothioate antagonist-bound forms using several 15N relaxation experiments. Based on the comparative analysis of dynamics in these three states, we have proposed a model of EPAC activation that incorporates the dynamic features allosterically modulated by cAMP and shows that cAMP binding weakens the IL by increasing its entropic penalty due to dynamic enhancements. PMID- 18411262 TI - Ganglioside inhibition of neurite outgrowth requires Nogo receptor function: identification of interaction sites and development of novel antagonists. AB - Gangliosides are key players in neuronal inhibition, with antibody-mediated clustering of gangliosides blocking neurite outgrowth in cultures and axonal regeneration post injury. In this study we show that the ganglioside GT1b can form a complex with the Nogo-66 receptor NgR1. The interaction is shown by analytical ultracentrifugation sedimentation and is mediated by the sialic acid moiety on GT1b, with mutations in FRG motifs on NgR1 attenuating the interaction. One FRG motif was developed into a cyclic peptide (N-AcCLQKFRGSSC-NH(2)) antagonist of GT1b, reversing the GT1b antibody inhibition of cerebellar granule cell neurite outgrowth. Interestingly, the peptide also antagonizes neurite outgrowth inhibition mediated by soluble forms of the myelin-associated glycoprotein (MAG). Structure function analysis of the peptide point to the conserved FRG triplet being the minimal functional motif, and mutations within this motif inhibit NgR1 binding to both GT1b and MAG. Finally, using gene ablation, we show that the cerebellar neuron response to GT1b antibodies and soluble MAG is indeed dependent on NgR1 function. The results suggest that gangliosides inhibit neurite outgrowth by interacting with FRG motifs in the NgR1 and that this interaction can also facilitate the binding of MAG to the NgR1. Furthermore, the results point to a rational strategy for developing novel ganglioside antagonists. PMID- 18411263 TI - The Legionella autoinducer synthase LqsA produces an alpha-hydroxyketone signaling molecule. AB - The opportunistic pathogen Legionella pneumophila replicates in human lung macrophages and in free-living amoebae. To accommodate the transfer between host cells, L. pneumophila switches from a replicative to a transmissive phase. L. pneumophila harbors a gene cluster homologous to the Vibrio cholerae cqsAS quorum sensing system, encoding a putative autoinducer synthase (lqsA) and a sensor kinase (lqsS), which flank a response regulator (lqsR). LqsR is an element of the L. pneumophila virulence regulatory network, which promotes pathogen-host cell interactions and inhibits entry into the replicative growth phase. Here, we show that lqsA functionally complements a V. cholerae cqsA autoinducer synthase deletion mutant and, upon expression in L. pneumophila or Escherichia coli, produces the diffusible signaling molecule LAI-1 (Legionella autoinducer-1). LAI 1 is distinct from CAI-1 (Cholerae autoinducer-1) and was identified as 3 hydroxypentadecan-4-one using liquid chromatography coupled to high resolution tandem mass spectrometry. The activity of both LqsA and CqsA was abolished upon mutation of a conserved lysine, and covalent binding of the cofactor pyridoxal 5' phosphate to this lysine was confirmed by mass spectrometry. Thus, LqsA and CqsA belong to a family of pyridoxal 5'-phosphate-dependent autoinducer synthases, which produce the alpha-hydroxyketone signaling molecules LAI-1 and CAI-1. PMID- 18411264 TI - Phosphorylation and stabilization of TAp63gamma by IkappaB kinase-beta. AB - Post-translational modification of the p53 family members is key to their regulation. Here we report the phosphorylation of TAp63gamma, but not DeltaNp63gamma, by IkappaB kinase beta (IKKbeta). Activation of IKKbeta by gamma radiation or tumor necrosis factor-alpha led to increased TAp63gamma protein levels in cells. IKKbeta, but not its kinase-defective mutant IKKbeta-K44A, led to this observed stabilization of TAp63gamma. This stabilization of TAp63gamma in response to gamma radiation was significantly decreased in the absence of IKKbeta. Phosphorylation of TAp63gamma blocks ubiquitylation and possible degradation of this protein. We postulate that phosphorylation of TAp63gamma by IKKbeta stabilizes the TAp63gamma protein by blocking ubiquitylation-dependent degradation of this protein. PMID- 18411265 TI - Interleukin 1alpha-induced NFkappaB activation and chemokine mRNA stabilization diverge at IRAK1. AB - Interleukin 1alpha (IL-1alpha) is capable of driving pro-inflammatory gene expression through both the initiation of transcription and by prolonging the half-life of short-lived mRNAs. Although the signaling events linking the IL-1 receptor to the activation of NFkappaB and the initiation of transcription have been well characterized, less is known about the signaling events linking to mRNA stabilization. As a model to study the control of mRNA stability we have used the mouse chemokine KC, expression of which requires both NFkappaB-driven transcription and stabilization of the constitutively unstable mRNA. We have evaluated the role of signaling adaptors known to play a role in IL-1alpha-driven NFkappaB activation in the generation of mRNA stability. Surprisingly, although TRAF6 is essential for NFkappaB activation, it is not required for IL-1alpha induced mRNA stabilization. IRAK1, which is recognized to function upstream of TRAF6, is required for both mRNA stabilization and activation of NFkappaB. Consistent with the previous findings, the TRAF6 interaction sites in IRAK1 are required for NFkappaB activation but do not play a role in mRNA stabilization. These findings indicate that signals from the IL-1 receptor segregate into at least two separate pathways at the level of IRAK1; one couples through TRAF6 to NFkappaB activation while a second utilizes a TRAF6-independent pathway that is responsible for mRNA stabilization. PMID- 18411266 TI - Splice-specific functions of gephyrin in molybdenum cofactor biosynthesis. AB - Gephyrin is a multifunctional protein involved in the clustering of inhibitory neuroreceptors. In addition, gephyrin catalyzes the last step in molybdenum cofactor (Moco) biosynthesis essential for the activities of Mo-dependent enzymes such as sulfite oxidase and xanthine oxidoreductase. Functional complexity and diversity of gephyrin is believed to be regulated by alternative splicing in a tissue-specific manner. Here, we investigated eight gephyrin variants with combinations of seven alternatively spliced exons located in the N-terminal G domain, the central domain, and the C-terminal E domain. Their activity in Moco synthesis was analyzed in vivo by reconstitution of gephyrin-deficient L929 cells, which were found to be defective in the G domain of gephyrin. Individual domain functions were assayed in addition and confirmed that variants containing either an additional C5 cassette or missing the C6 cassette are inactive in Moco synthesis. In contrast, different alterations within the central domain retained the Moco synthetic activity of gephyrin. The recombinant gephyrin G domain containing the C5 cassette forms dimers in solution, binds molybdopterin, but is unable to catalyze molybdopterin (MPT) adenylylation. Determination of Moco and MPT content in different tissues showed that besides liver and kidney, brain was capable of synthesizing Moco most efficiently. Subsequent analysis of cultured neurons and glia cells demonstrated glial Moco synthesis due to the expression of gephyrins containing the cassettes C2 and C6 with and without C3.1. PMID- 18411267 TI - Decreased ceramide transport protein (CERT) function alters sphingomyelin production following UVB irradiation. AB - Increased cellular ceramide accounts in part for UVB irradiation-induced apoptosis in cultured human keratinocytes with concurrent increased glucosylceramide but not sphingomyelin generation in these cells. Given that conversion of ceramide to non-apoptotic metabolites such as sphingomyelin and glucosylceramide protects cells from ceramide-induced apoptosis, we hypothesized that failed up-regulation of sphingomyelin generation contributes to ceramide accumulation following UVB irradiation. Because both sphingomyelin synthase and glucosylceramide synthase activities were significantly decreased in UVB irradiated keratinocytes, we investigated whether alteration(s) in the function of ceramide transport protein (or CERT) required for sphingomyelin synthesis occur(s) in UVB-irradiated cells. Fluorescently labeled N-(4,4-difluoro-5,7 dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-d-erythro-sphingosine (C(5) DMB-ceramide) relocation to the Golgi was diminished after irradiation, consistent with decreased CERT function, whereas the CERT inhibitor N-(3-hydroxy 1-hydroxymethyl-3-phenylpropyl)dodecanamide (1R,3R isomer) (HPA-12) produced an equivalent effect. UVB irradiation also induced the rapid formation of a stable CERT homotrimer complex in keratinocytes as determined by Western immunoblot and mass spectrometry analyses, a finding replicated in HeLa, HEK293T, and HaCaT cells and in murine epidermis. Ceramide binding activity was decreased in recombinant CERT proteins containing the UVB-induced homotrimer. The middle region domain of the CERT protein was required for the homotrimer formation, whereas neither the pleckstrin homology (Golgi-binding) nor the START (ceramide binding) domains were involved. Finally like UVB-treated keratinocytes, HPA-12 blockade of CERT function increased keratinocyte apoptosis, decreased sphingomyelin synthesis, and led to accumulation of ceramide. Thus, UVB-induced CERT homotrimer formation accounts, at least in part, for apoptosis and failed up regulation of sphingomyelin synthesis following UVB irradiation, revealing that inactive CERT can attenuate a key metabolic protective mechanism against ceramide induced apoptosis in keratinocytes. PMID- 18411268 TI - Identification of a new urate and high affinity nicotinate transporter, hOAT10 (SLC22A13). AB - The orphan transporter hORCTL3 (human organic cation transporter like 3; SLC22A13) is highly expressed in kidneys and to a weaker extent in brain, heart, and intestine. hORCTL3-expressing Xenopus laevis oocytes showed uptake of [(3)H]nicotinate, [(3)H]p-aminohippurate, and [(14)C]urate. Hence, hORCTL3 is an organic anion transporter, and we renamed it hOAT10. [(3)H]Nicotinate transport by hOAT10 into X. laevis oocytes and into Caco-2 cells was saturable with Michaelis constants (K(m)) of 22 and 44 microm, respectively, suggesting that hOAT10 may be the molecular equivalent of the postulated high affinity nicotinate transporter in kidneys and intestine. The pH dependence of hOAT10 suggests p aminohippurate(-)/OH(-), urate(-)/OH(-), and nicotinate(-)/OH(-) exchange as possible transport modes. Urate inhibited [(3)H]nicotinate transport by hOAT10 with an IC(50) value of 759 microm, assuming that hOAT10 represents a low affinity urate transporter. hOAT10-mediated [(14)C]urate uptake was elevated by an exchange with l -lactate, pyrazinoate, and nicotinate. Surprisingly, we have detected urate(-)/glutathione exchange by hOAT10, consistent with an involvement of hOAT10 in the renal glutathione cycle. Uricosurics, diuretics, and cyclosporine A showed substantial interactions with hOAT10, of which cyclosporine A enhanced [(14)C]urate uptake, providing the first molecular evidence for cyclosporine A-induced hyperuricemia. PMID- 18411269 TI - Structure and function of LGP2, a DEX(D/H) helicase that regulates the innate immunity response. AB - RNA recognition receptors are important for detection of and response to viral infections. RIG-I and MDA5 are cytoplasmic DEX(D/H) helicase proteins that can induce signaling in response to RNA ligands, including those from viral infections. LGP2, a homolog of RIG-I and MDA5 without the caspase recruitment domain required for signaling, plays an important role in modulating signaling by MDA5 and RIG-I, presumably through heterocomplex formation and/or by serving as a sink for RNAs. Here we demonstrate that LGP2 can be coexpressed with RIG-I to inhibit activation of the NF-kappaB reporter expression and that LGP2 protein produced in insect cells can bind both single- and double-stranded RNA (dsRNA), with higher affinity and cooperativity for dsRNA. Electron microscopy and image reconstruction were used to determine the shape of the LGP2 monomer in the absence of dsRNA and of the dimer complexed to a 27-bp dsRNA. LGP2 has striking structural similarity to the helicase domain of the superfamily 2 DNA helicase, Hef. PMID- 18411271 TI - Substrate specificity and membrane topology of Escherichia coli PgpB, an undecaprenyl pyrophosphate phosphatase. AB - The synthesis of the lipid carrier undecaprenyl phosphate (C(55)-P) requires the dephosphorylation of its precursor, undecaprenyl pyrophosphate (C(55)-PP). The latter lipid is synthesized de novo in the cytosol and is also regenerated after its release from the C(55)-PP-linked glycans in the periplasm. In Escherichia coli the dephosphorylation of C(55)-PP was shown to involve four integral membrane proteins, BacA, and three members of the type 2 phosphatidic acid phosphatase family, PgpB, YbjG, and YeiU. Here, the PgpB protein was purified to homogeneity, and its phosphatase activity was examined. This enzyme was shown to catalyze the dephosphorylation of C(55)-PP with a relatively low efficiency compared with diacylglycerol pyrophosphate and farnesyl pyrophosphate (C(15)-PP) lipid substrates. However, the in vitro C(55)-PP phosphatase activity of PgpB was specifically enhanced by different phospholipids. We hypothesize that the phospholipids are important determinants to ensure proper conformation of the atypical long axis C(55) carrier lipid in membranes. Furthermore, a topological analysis demonstrated that PgpB contains six transmembrane segments, a large periplasmic loop, and the type 2 phosphatidic acid phosphatase signature residues at a periplasmic location. PMID- 18411270 TI - Prosegment of tripeptidyl peptidase I is a potent, slow-binding inhibitor of its cognate enzyme. AB - Tripeptidyl peptidase I (TPP I) is the first mammalian representative of a family of pepstatin-insensitive serine-carboxyl proteases, or sedolisins. The enzyme acts in lysosomes, where it sequentially removes tripeptides from the unmodified N terminus of small, unstructured polypeptides. Naturally occurring mutations in TPP I underlie a neurodegenerative disorder of childhood, classic late infantile neuronal ceroid lipofuscinosis (CLN2). Generation of mature TPP I is associated with removal of a long prosegment of 176 amino acid residues from the zymogen. Here we investigated the inhibitory properties of TPP I prosegment expressed and isolated from Escherichia coli toward its cognate protease. We show that the TPP I prosegment is a potent, slow-binding inhibitor of its parent enzyme, with an overall inhibition constant in the low nanomolar range. We also demonstrate the protective effect of the prosegment on alkaline pH-induced inactivation of the enzyme. Interestingly, the inhibitory properties of TPP I prosegment with the introduced classic late infantile neuronal ceroid lipofuscinosis disease associated mutation, G77R, significantly differed from those revealed by wild type prosegment in both the mechanism of interaction and the inhibitory rate. This is the first characterization of the inhibitory action of the sedolisin prosegment. PMID- 18411272 TI - A common theme in interaction of bacterial immunoglobulin-binding proteins with immunoglobulins illustrated in the equine system. AB - The M protein of Streptococcus equi subsp. equi known as fibrinogen-binding protein (FgBP) is a cell wall-associated protein with antiphagocytic activity that binds IgG. Recombinant versions of the seven equine IgG subclasses were used to investigate the subclass specificity of FgBP. FgBP bound predominantly to equine IgG4 and IgG7, with little or no binding to the other subclasses. Competitive binding experiments revealed that FgBP could inhibit the binding of staphylococcal protein A and streptococcal protein G to both IgG4 and IgG7, implicating the Fc interdomain region in binding to FgBP. To identify which of the two IgG Fc domains contributed to the interaction with FgBP, we tested two human IgG1/IgA1 domain swap mutants and found that both domains are required for full binding, with the CH3 domain playing a critical role. The binding site for FgBP was further localized using recombinant equine IgG7 antibodies with single or double point mutations to residues lying at the CH2-CH3 interface. We found that interaction of FgBP with equine IgG4 and IgG7 was able to disrupt C1q binding and antibody-mediated activation of the classical complement pathway, demonstrating an effective means by which S. equi may evade the immune response. The mode of interaction of FgBP with IgG fits a common theme for bacterial Ig binding proteins. Remarkably, for those interactions studied in detail, it emerges that all the Ig-binding proteins target the CH2-CH3 domain interface, regardless of specificity for IgG or IgA, streptococcal or staphylococcal origin, or host species (equine or human). PMID- 18411273 TI - Expression of the novel wheat gene TM20 confers enhanced cadmium tolerance to bakers' yeast. AB - Cadmium causes the generation of reactive oxygen species, which in turn causes cell damage. We isolated a novel gene from a wheat root cDNA library, which conferred Cd(II)-specific tolerance when expressed in yeast (Saccharomyces cerevisiae). The gene, which we called TaTM20, for Triticum aestivum transmembrane 20, encodes a putative hydrophobic polypeptide of 889 amino acids, containing 20 transmembrane domains arranged as a 5-fold internal repeating unit of 4 transmembrane domains each. Expression of TaTM20 in yeast cells stimulated Cd(II) efflux resulting in a decrease in the content of yeast intracellular cadmium. TaTM20-induced Cd(II) tolerance was maintained in yeast even under conditions of reduced GSH. These results demonstrate that TaTM20 enhances Cd(II) tolerance in yeast through the stimulation of Cd(II) efflux from the cell, partially independent of GSH. Treatment of wheat seedlings with Cd(II) induced their expression of TaTM20, decreasing subsequent root Cd(II) accumulation and suggesting a possible role for TaTM20 in Cd(II) tolerance in wheat. PMID- 18411274 TI - The in vitro RNA synthesizing activity of the isolated arterivirus replication/transcription complex is dependent on a host factor. AB - The cytoplasmic replication of positive-stranded RNA viruses is associated with characteristic, virus-induced membrane structures that are derived from host cell organelles. We used the prototype arterivirus, equine arteritis virus (EAV), to gain insight into the structure and function of the replication/transcription complex (RTC) of nidoviruses. RTCs were isolated from EAV-infected cells, and their activity was studied using a newly developed in vitro assay for viral RNA synthesis, which reproduced the synthesis of both viral genome and subgenomic mRNAs. A detailed characterization of this system and its reaction products is described. RTCs isolated from cytoplasmic extracts by differential centrifugation were inactive unless supplemented with a cytosolic host protein factor, which, according to subsequent size fractionation analysis, has a molecular mass in the range of 59-70 kDa. This host factor was found to be present in a wide variety of eukaryotes. Several EAV replicase subunits cosedimented with newly made viral RNA in a heavy membrane fraction that contained all RNA-dependent RNA polymerase activity. This fraction contained the characteristic double membrane vesicles (DMVs) that were previously implicated in EAV RNA synthesis and could be immunolabeled for EAV nonstructural proteins (nsps). Replicase subunits directly involved in viral RNA synthesis (nsp9 and nsp10) or DMV formation (nsp2 and nsp3) exclusively cosedimented with the active RTC. Subgenomic mRNAs appeared to be released from the complex, whereas newly made genomic RNA remained more tightly associated. Taken together, our data strongly support a link between DMVs and the RNA-synthesizing machinery of arteriviruses. PMID- 18411275 TI - The catalytic domain of insulin-degrading enzyme forms a denaturant-resistant complex with amyloid beta peptide: implications for Alzheimer disease pathogenesis. AB - Insulin-degrading enzyme (IDE) is central to the turnover of insulin and degrades amyloid beta (Abeta) in the mammalian brain. Biochemical and genetic data support the notion that IDE may play a role in late onset Alzheimer disease (AD), and recent studies suggest an association between AD and diabetes mellitus type 2. Here we show that a natively folded recombinant IDE was capable of forming a stable complex with Abeta that resisted dissociation after treatment with strong denaturants. This interaction was also observed with rat brain IDE and detected in an SDS-soluble fraction from AD cortical tissue. Abeta sequence 17-27, known to be crucial in amyloid assembly, was sufficient to form a stable complex with IDE. Monomeric as opposed to aggregated Abeta was competent to associate irreversibly with IDE following a very slow kinetics (t(1/2) approximately 45 min). Partial denaturation of IDE as well as preincubation with a 10-fold molar excess of insulin prevented complex formation, suggesting that the irreversible interaction of Abeta takes place with at least part of the substrate binding site of the protease. Limited proteolysis showed that Abeta remained bound to a approximately 25-kDa N-terminal fragment of IDE in an SDS-resistant manner. Mass spectrometry after in gel digestion of the IDE .Abeta complex showed that peptides derived from the region that includes the catalytic site of IDE were recovered with Abeta. Taken together, these results are suggestive of an unprecedented mechanism of conformation-dependent substrate binding that may perturb Abeta clearance, insulin turnover, and promote AD pathogenesis. PMID- 18411276 TI - Leukotriene E4 activates peroxisome proliferator-activated receptor gamma and induces prostaglandin D2 generation by human mast cells. AB - Cysteinyl leukotrienes (cys-LTs) are potent inflammatory lipid mediators, of which leukotriene (LT) E(4) is the most stable and abundant in vivo. Although only a weak agonist of established G protein-coupled receptors (GPCRs) for cys LTs, LTE(4) potentiates airway hyper-responsiveness (AHR) by a cyclooxygenase (COX)-dependent mechanism and induces bronchial eosinophilia. We now report that LTE(4) activates human mast cells (MCs) by a pathway involving cooperation between an MK571-sensitive GPCR and peroxisome proliferator-activated receptor (PPAR)gamma, a nuclear receptor for dietary lipids. Although LTD(4) is more potent than LTE(4) for inducing calcium flux by the human MC sarcoma line LAD2, LTE(4) is more potent for inducing proliferation and chemokine generation, and is at least as potent for upregulating COX-2 expression and causing prostaglandin D(2) (PGD(2)) generation. LTE(4) caused phosphorylation of extracellular signal regulated kinase (ERK), p90RSK, and cyclic AMP-regulated-binding protein (CREB). ERK activation in response to LTE(4), but not to LTD(4), was resistant to inhibitors of phosphoinositol 3-kinase. LTE(4)-mediated COX-2 induction, PGD(2) generation, and ERK phosphorylation were all sensitive to interference by the PPARgamma antagonist GW9662 and to targeted knockdown of PPARgamma. Although LTE(4)-mediated PGD(2) production was also sensitive to MK571, an antagonist for the type 1 receptor for cys-LTs (CysLT(1)R), it was resistant to knockdown of this receptor. This LTE(4)-selective receptor-mediated pathway may explain the unique physiologic responses of human airways to LTE(4) in vivo. PMID- 18411277 TI - The miR-200 family inhibits epithelial-mesenchymal transition and cancer cell migration by direct targeting of E-cadherin transcriptional repressors ZEB1 and ZEB2. AB - MicroRNAs are small non-coding RNA molecules that can regulate gene expression by interacting with multiple mRNAs and inducing either translation suppression or degradation of mRNA. Recently, several miRNAs were identified as either promoters or suppressors of metastasis. However, it is unclear in which step(s) of the multistep metastatic cascade these miRNAs play a defined functional role. To study the functional importance of miRNAs in epithelial-mesenchymal transition (EMT), a process thought to initiate metastasis by enhancing the motility of tumor cells, we used a well established in vitro EMT assay: transforming growth factor-beta-induced EMT in NMuMG murine mammary epithelial cells. We found that members of the miR-200 family, organized as two clusters in the genome, were repressed during EMT. Overexpression of each miRNA individually or as clusters in NMuMG cells hindered EMT by enhancing E-cadherin expression through direct targeting of ZEB1 and ZEB2, which encode transcriptional repressors of E cadherin. In the 4TO7 mouse carcinoma cell line, which expresses low levels of endogenous E-cadherin and displays a mesenchymal phenotype, ectopic expression of the miR-200 family miRNAs significantly increased E-cadherin expression and altered cell morphology to an epithelial phenotype. Furthermore, ectopic expression of each miR-200 miRNA cluster significantly reduced the in vitro motility of 4TO7 cells in migration assays. These results suggested that loss of expression of the miR-200 family members may play a critical role in the repression of E-cadherin by ZEB1 and ZEB2 during EMT, thereby enhancing migration and invasion during cancer progression. PMID- 18411278 TI - New Determinant for the CaVbeta2 subunit modulation of the CaV1.2 calcium channel. AB - Ca(v)beta subunits support voltage gating of Ca(v)1.2 calcium channels and play important role in excitation-contraction coupling. The common central membrane associated guanylate kinase (MAGUK) region of Ca(v)beta binds to the alpha interaction domain (AID) and the IQ motif of the pore-forming alpha(1C) subunit, but these two interactions do not explain why the cardiac Ca(v)beta(2) subunit splice variants differentially modulate inactivation of Ca(2+) currents (I(Ca)). Previously we described beta(2Deltag), a functionally active splice variant of human Ca(v)beta(2) lacking MAGUK. By deletion analysis of beta(2Deltag), we have now identified a 41-amino acid C-terminal essential determinant (beta(2)CED) that stimulates I(Ca) in the absence of Ca(v)beta subunits and conveys a +20-mV shift in the peak of the I(Ca)-voltage relationship. The beta(2)CED is targeted by alpha(1C) to the plasma membrane, forms a complex with alpha(1C) but does not bind to AID. Electrophysiology and binding studies point to the calmodulin interacting LA/IQ region in the alpha(1C) subunit C terminus as a functionally relevant beta(2)CED binding site. The beta(2)CED interacts with LA/IQ in a Ca(2+) and calmodulin-independent manner and need LA, but not IQ, to activate the channel. Deletion/mutation analyses indicated that each of the three Ca(v)beta(2)/alpha(1C) interactions is sufficient to support I(Ca). However, beta(2)CED does not support Ca(2+)-dependent inactivation, suggesting that interactions of MAGUK with AID and IQ are crucial for Ca(2+)-induced inactivation. The beta(2)CED is conserved only in Ca(v)beta(2) subunits. Thus, beta(2)CED constitutes a previously unknown integrative part of the multifactorial mechanism of Ca(v)beta(2)-subunit differential modulation of the Ca(v)1.2 calcium channel that in beta(2Deltag) occurs without MAGUK. PMID- 18411280 TI - Control of the timing of promoter escape and RNA catalysis by the transcription factor IIb fingertip. AB - Transcription factor IIB (TFIIB) recruits RNA polymerase II to promoters and inserts a finger domain into its active site, with unknown consequences. Here we show that that the tip of this finger is important for two transcription initiation functions. First, TFIIB acts as a catalytic cofactor for initial RNA bond formation. It does so via a pair of fingertip aspartates that can bind magnesium, placing TFIIB within a family of proteins that insert finger domains to alter the catalytic functions of RNA polymerase. Second, the TFIIB fingertip mediates the timing of the release of TFIIB that is associated with appropriate promoter escape. These initiation requirements may assist in RNA quality control by minimizing functional synthesis when RNA polymerase becomes inappropriately associated with the genome without having been recruited there by TFIIB. PMID- 18411279 TI - Regulation of glycan structures in animal tissues: transcript profiling of glycan related genes. AB - Glycan structures covalently attached to proteins and lipids play numerous roles in mammalian cells, including protein folding, targeting, recognition, and adhesion at the molecular or cellular level. Regulating the abundance of glycan structures on cellular glycoproteins and glycolipids is a complex process that depends on numerous factors. Most models for glycan regulation hypothesize that transcriptional control of the enzymes involved in glycan synthesis, modification, and catabolism determines glycan abundance and diversity. However, few broad-based studies have examined correlations between glycan structures and transcripts encoding the relevant biosynthetic and catabolic enzymes. Low transcript abundance for many glycan-related genes has hampered broad-based transcript profiling for comparison with glycan structural data. In an effort to facilitate comparison with glycan structural data and to identify the molecular basis of alterations in glycan structures, we have developed a medium-throughput quantitative real time reverse transcriptase-PCR platform for the analysis of transcripts encoding glycan-related enzymes and proteins in mouse tissues and cells. The method employs a comprehensive list of >700 genes, including enzymes involved in sugar-nucleotide biosynthesis, transporters, glycan extension, modification, recognition, catabolism, and numerous glycosylated core proteins. Comparison with parallel microarray analyses indicates a significantly greater sensitivity and dynamic range for our quantitative real time reverse transcriptase-PCR approach, particularly for the numerous low abundance glycan related enzymes. Mapping of the genes and transcript levels to their respective biosynthetic pathway steps allowed a comparison with glycan structural data and provides support for a model where many, but not all, changes in glycan abundance result from alterations in transcript expression of corresponding biosynthetic enzymes. PMID- 18411281 TI - Signaling and cross-talk by C5a and UDP in macrophages selectively use PLCbeta3 to regulate intracellular free calcium. AB - Studies in fibroblasts, neurons, and platelets have demonstrated the integration of signals from different G protein-coupled receptors (GPCRs) in raising intracellular free Ca(2+). To study signal integration in macrophages, we screened RAW264.7 cells and bone marrow-derived macrophages (BMDM) for their Ca(2+) response to GPCR ligands. We found a synergistic response to complement component 5a (C5a) in combination with uridine 5'-diphosphate (UDP), platelet activating factor (PAF), or lysophosphatidic acid (LPA). The C5a response was Galpha(i)-dependent, whereas the UDP, PAF, and LPA responses were Galpha(q) dependent. Synergy between C5a and UDP, mediated by the C5a and P2Y6 receptors, required dual receptor occupancy, and affected the initial release of Ca(2+) from intracellular stores as well as sustained Ca(2+) levels. C5a and UDP synergized in generating inositol 1,4,5-trisphosphate, suggesting synergy in activating phospholipase C (PLC) beta. Macrophages expressed transcripts for three PLCbeta isoforms (PLCbeta2, PLCbeta3, and PLCbeta4), but GPCR ligands selectively used these isoforms in Ca(2+) signaling. C5a predominantly used PLCbeta3, whereas UDP used PLCbeta3 but also PLCbeta4. Neither ligand required PLCbeta2. Synergy between C5a and UDP likewise depended primarily on PLCbeta3. Importantly, the Ca(2+) signaling deficiency observed in PLCbeta3-deficient BMDM was reversed by re-constitution with PLCbeta3. Neither phosphatidylinositol (PI) 3-kinase nor protein kinase C was required for synergy. In contrast to Ca(2+), PI 3-kinase activation by C5a was inhibited by UDP, as was macropinocytosis, which depends on PI 3-kinase. PLCbeta3 may thus provide a selective target for inhibiting Ca(2+) responses to mediators of inflammation, including C5a, UDP, PAF, and LPA. PMID- 18411282 TI - Phosphorylation by casein kinase 1 regulates tonicity-induced osmotic response element-binding protein/tonicity enhancer-binding protein nucleocytoplasmic trafficking. AB - The osmotic response element-binding protein (OREBP), also known as tonicity enhancer-binding protein (TonEBP) or NFAT5, is the only known osmo-sensitive transcription factor that mediates cellular adaptations to extracellular hypertonic stress. Although it is well documented that the subcellular localization and transactivation activity of OREBP/TonEBP are tightly regulated by extracellular tonicity, the molecular mechanisms involved remain elusive. Here we show that nucleocytoplasmic trafficking of OREBP/TonEBP is regulated by the dual phosphorylation of Ser-155 and Ser-158. Alanine scanning mutagenesis revealed that Ser-155 is an essential residue that regulates OREBP/TonEBP nucleocytoplasmic trafficking. Tandem mass spectrometry revealed that Ser-155 and Ser-158 of OREBP/TonEBP are both phosphorylated in living cells under hypotonic conditions. In vitro phosphorylation assays further suggest that phosphorylation of the two serine residues proceeds in a hierarchical manner with phosphorylation of Ser-155 priming the phosphorylation of Ser-158 and that these phosphorylations are essential for nucleocytoplasmic trafficking of the transcription factor. Finally, we have shown that the pharmacological inhibition of casein kinase 1 (CK1) abolishes the phosphorylation of Ser-158 and impedes OREBP/TonEBP nuclear export and that recombinant CK1 phosphorylates Ser-158. Knockdown of CK1alpha1L, a novel isoform of CK1, inhibits hypotonicity-induced OREBP/TonEBP nuclear export. Together these data highlight the importance of Ser-155 and Ser-158 in the nucleocytoplasmic trafficking of OREBP/TonEBP and indicate that CK1 plays a major role in regulating this process. PMID- 18411283 TI - Matrix metalloproteinase processing of CXCL11/I-TAC results in loss of chemoattractant activity and altered glycosaminoglycan binding. AB - The CXCR3 chemokine receptor regulates the migration of Th1 lymphocytes and responds to three ligands: CXCL9/MIG, CXCL10/IP-10, and CXCL11/I-TAC. We screened for potential regulation of T cell responses by matrix metalloproteinase (MMP) processing of these important chemokines. The most potent of the CXCR3 ligands, CXCL11, was identified by matrix-assisted laser desorption ionization time-of flight mass spectrometry as a substrate of the PMN-specific MMP-8, macrophage specific MMP-12, and the general leukocyte MMP-9. The 73-amino acid residue CXCL11 is processed at both the amino and carboxyl termini to generate CXCL11-(5 73), -(5-63), and -(5-58) forms. NH2-terminal truncation results in loss of agonistic properties, as shown in calcium mobilization and chemotaxis experiments using CXCR3 transfectants and human T lymphocytes. Moreover, CXCL11-(5-73) is a CXCR3 antagonist and interestingly shows enhanced affinity to heparin. However, upon COOH-terminal truncation to position 58 there is loss of antagonist activity and heparin binding. Together this highlights an unexpected site for receptor interaction and that the carboxyl terminus is critical for glycosaminoglycan binding, an essential function for the formation of chemokine gradients in vivo. Hence, MMP activity might regulate CXCL11 tissue gradients in two ways. First, the potential of CXCL11-(5-73) to compete active CXCL11 from glycosaminoglycans might lead to the formation of an antagonistic haptotactic chemokine gradient. Second, upon further truncation, MMPs disperse the CXCL11 gradients in a novel way by proteolytic loss of a COOH-terminal GAG binding site. Hence, these results reveal potential new roles in down-regulating Th1 lymphocyte chemoattraction through MMP processing of CXCL11. PMID- 18411284 TI - Inhibition of Neisseria gonorrhoeae epithelial cell interactions by vaginal Lactobacillus species. AB - High levels of Lactobacillus, the dominant genus of the healthy human vaginal microbiota, have been epidemiologically linked to a reduced risk of infection following exposure to the sexually transmitted pathogen Neisseria gonorrhoeae. In this work, a cell culture model of gonococcal infection was adapted to examine the effects of lactobacilli on gonococcal interactions with endometrial epithelial cells in vitro. Precolonization of epithelial cells with Lactobacillus jensenii, Lactobacillus gasseri ATCC 33323, or L. gasseri ATCC 9857 reduced gonococcal adherence by nearly 50%. Lactobacilli also inhibited gonococcal invasion of epithelial cells by more than 60%, which was independent of the effect on adherence. Furthermore, lactobacilli were able to displace adherent gonococci from epithelial cells, suggesting that these organisms have potential as a postexposure prophylactic. Thus, vaginal lactobacilli have the ability to inhibit gonococci at two key steps of an infection, which might have a significant effect in determining whether the gonococcus will be able to successfully establish an infection following exposure in vivo. PMID- 18411285 TI - Utilization of an intracellular bacterial community pathway in Klebsiella pneumoniae urinary tract infection and the effects of FimK on type 1 pilus expression. AB - Klebsiella pneumoniae is an important cause of urinary tract infection (UTI), but little is known about its pathogenesis in vivo. The pathogenesis of the K. pneumoniae cystitis isolate TOP52 was compared to that of the uropathogenic Escherichia coli (UPEC) isolate UTI89 in a murine cystitis model. Bladder and kidney titers of TOP52 were lower than those of UTI89 at early time points but similar at later time points. TOP52, like UTI89, formed biofilm-like intracellular bacterial communities (IBCs) within the murine bladder, albeit at significantly lower levels than UTI89. Additionally, filamentation of TOP52 was observed, a process critical for UTI89 evasion of neutrophil phagocytosis and persistence in the bladder. Thus, the IBC pathway is not specific to UPEC alone. We investigated if differences in type 1 pilus expression may explain TOP52's early defect in vivo. The type 1 pilus operon is controlled by recombinase mediated (fimE, fimB, and fimX) phase variation of an invertible promoter element. We found that K. pneumoniae carries an extra gene of unknown function at the 3' end of its type 1 operon, fimK, and the genome lacks the recombinase fimX. A deletion mutant of fimK was constructed, and TOP52 Delta fimK had higher titers and formed more IBCs in the murine cystitis model than wild type. The loss of fimK or expression of E. coli fimX from a plasmid in TOP52 resulted in a larger phase-ON population and higher expression levels of type 1 pili and gave TOP52 the ability to form type 1-dependent biofilms. Complementation with pfimK decreased type 1 pilus expression and biofilm formation of TOP52 Delta fimK and decreased UTI89 biofilm formation. Thus, K. pneumoniae appears programmed for minimal expression of type 1 pili, which may explain, in part, why K. pneumoniae is a less prevalent etiologic agent of UTI than UPEC. PMID- 18411287 TI - Beta-hydroxybutyrate abrogates formation of bovine neutrophil extracellular traps and bactericidal activity against mammary pathogenic Escherichia coli. AB - Escherichia coli is an important bacterial species isolated from bovine mastitis. The rate of neutrophil recruitment into the mammary gland and their bactericidal activity largely affect the severity and outcome of the disease. Ketosis is a common metabolic disease, and affected dairy cows are known to have increased risk for mastitis and other infectious conditions. The disease is associated with high blood and milk levels of beta-hydroxybutyrate (BHBA), previously shown to negatively affect neutrophil function by unknown mechanisms. We show here that the mammary pathogenic E. coli strain P4 activates normal bovine neutrophils to form neutrophil extracellular traps (NETs), which are highly bactericidal against this organism. Preincubation of these neutrophils with increasing concentrations (0.1 to 8 mmol/liter) of BHBA caused a fivefold decrease of E. coli P4 phagocytosis, though intracellular killing was unaffected. Furthermore, BHBA caused a 10-fold decrease in the NETs formed by E. coli P4-activated neutrophils and a similar decrease in NET bactericidal activity against this organism. These negative effects of BHBA on bovine neutrophils might explain the increased susceptibility of ketotic cows to mastitis and other infectious conditions. PMID- 18411286 TI - Multiple roles of phospholipase A2 during lung infection and inflammation. PMID- 18411288 TI - Nasal vaccination with the 40-kilodalton outer membrane protein of Porphyromonas gingivalis and a nontoxic chimeric enterotoxin adjuvant induces long-term protective immunity with reduced levels of immunoglobulin E antibodies. AB - In this study, we demonstrated that the 40-kDa outer membrane protein of Porphyromonas gingivalis (40-kDa OMP) nasally administered with a nontoxic chimeric adjuvant that combines the A subunit of mutant cholera toxin E112K with the pentameric B subunit of heat-labile enterotoxin from enterotoxigenic Escherichia coli (mCTA/LTB) elicited a long-term protective immune response. Immunization with the 40-kDa OMP and mCTA/LTB induced high levels of 40-kDa-OMP specific immunoglobulin G (IgG) and IgA antibodies (Abs) in sera and elicited a significant IgA anti-40-kDa OMP Ab response in saliva. These Ab responses were maintained for at least 1 year after the immunization. Although using adjuvant mCTA/LTB gave Ab responses in the saliva comparable to those obtained using native cholera toxin (nCT) as the adjuvant, the levels of total IgE and 40-kDa OMP-specific IgE Abs as well as interleukin-4 levels induced by the immunization with mCTA/LTB were lower than those induced by the immunization with nCT. Importantly, IgG Abs generated by nasal immunization with the 40-kDa OMP plus mCTA/LTB inhibited the coaggregation and hemagglutinin activities of P. gingivalis. Furthermore, the mice given nasal 40-kDa OMP plus mCTA/LTB showed a significant reduction of alveolar bone loss caused by oral infection with P. gingivalis even 1 year after the immunization compared to the loss in unimmunized mice. Because mCTA/LTB is nontoxic, nasally administered 40-kDa OMP together with mCTA/LTB should be an effective and safe mucosal vaccine against P. gingivalis infection in humans and may be an important tool for the prevention of chronic periodontitis. PMID- 18411289 TI - Role for myosin II in regulating positioning of Salmonella-containing vacuoles and intracellular replication. AB - Salmonella enterica serovar Typhimurium grows within host cells in a permissive compartment termed the Salmonella-containing vacuole (SCV). These bacteria use two distinct type III secretion systems (T3SS) to deliver virulence proteins (effectors) into cells. Effectors secreted by the Salmonella pathogenicity island 1 (SPI-1)-encoded T3SS mediate invasion and early SCV maturation steps, while those secreted by the SPI-2 T3SS affect the SCV at later stages postinfection. Some SPI-2 effectors modulate microtubule motor activity on the SCV. Here, we show that the actin-based motor myosin II also affects SCV dynamics during infection. Following invasion, myosin II is required for SCV positioning near the nucleus of host cells. Later, myosin II counteracts the activities of the SPI-2 effectors PipB2 and SseJ to maintain SCV positioning and stability, respectively. Myosin II activity was required for maximal bacterial growth in macrophages. Rho kinase activity was required for SCV positioning. The effector SopB, a known activator of Rho GTPases, was found to be required for SCV positioning, and transfection of cells with SopB was sufficient to induce myosin II phosphorylation. These studies reveal a novel role for myosin II in controlling SCV dynamics during infection and suggest that SopB activates myosin II. PMID- 18411290 TI - Protective effect of an extract from Ascaris suum in experimental arthritis models. AB - We investigated the effect of an extract from a helminth (Ascaris suum) in zymosan-induced arthritis (ZYA) or collagen-induced arthritis (CIA). Rats and mice, respectively, received 1 mg and 0.1 mg zymosan intra-articularly (i.a.). Test groups received an A. suum extract either per os (p.o.) or intraperitoneally (i.p.) 30 min prior to i.a. zymosan. Controls received saline. Hypernociception was measured using the articular incapacitation test. Cell influx, nitrite, and cytokine levels were assessed in joint exudates. The synovia and distal femoral extremities were used for histopathology. Cartilage damage was assessed through determining glycosaminoglycan (GAG) content. DBA/1J mice were subjected to CIA. The test group received A. suum extract i.p. 1 day after CIA became clinically detectable. Clinical severity and hypernociception were assessed daily. Neutrophil influx was determined using myeloperoxidase activity. The A. suum extract, either i.p. or p.o., significantly and dose-dependently inhibited cell influx and hypernociception in ZYA in addition to reducing GAG loss and ameliorating synovitis. The A. suum extract reduced i.a. levels of NO, interleukin-1beta (IL-1beta), and IL-10 but not tumor necrosis factor alpha (TNF alpha) in rats subjected to ZYA while reducing i.a. IL-10, but not IL-1beta or TNF-alpha, levels in mice. Clinically, mice subjected to CIA treated with the A. suum extract had less severe arthritis. Hypernociception, myeloperoxidase activity, and synovitis severity were significantly reduced. These data show that a helminth extract given p.o. protects from arthritis severity in two classical arthritis models. This A. suum effect is species independent and functions orally and parenterally. The results show clinical and structural benefits when A. suum extract is given either prophylactically or therapeutically. PMID- 18411291 TI - gp96 is a human colonocyte plasma membrane binding protein for Clostridium difficile toxin A. AB - Clostridium difficile toxin A (TxA), a key mediator of antibiotic-associated colitis, requires binding to a cell surface receptor prior to internalization. Our aim was to identify novel plasma membrane TxA binding proteins on human colonocytes. TxA was coupled with biotin and cross-linked to the surface of HT29 human colonic epithelial cells. The main colonocyte binding protein for TxA was identified as glycoprotein 96 (gp96) by coimmunoprecipitation and mass spectrum analysis. gp96 is a member of the heat shock protein family, which is expressed on human colonocyte apical membranes as well as in the cytoplasm. TxA binding to gp96 was confirmed by fluorescence immunostaining and in vitro coimmunoprecipitation. Following TxA binding, the TxA-gp96 complex was translocated from the cell membrane to the cytoplasm. Pretreatment with gp96 antibody decreased TxA binding to colonocytes and inhibited TxA-induced cell rounding. Small interfering RNA directed against gp96 reduced gp96 expression and cytotoxicity in colonocytes. TxA-induced inflammatory signaling via p38 and apoptosis as measured by activation of BAK (Bcl-2 homologous antagonist/killer) and DNA fragmentation were decreased in gp96-deficient B cells. We conclude that human colonocyte gp96 serves as a plasma membrane binding protein that enhances cellular entry of TxA, participates in cellular signaling events in the inflammatory cascade, and facilitates cytotoxicity. PMID- 18411292 TI - Helicobacter hepaticus infection promotes colon tumorigenesis in the BALB/c-Rag2( /-) Apc(Min/+) mouse. AB - Adenomatous polyposis coli (APC) mutations are linked to human and mouse colorectal cancers. The Apc multiple intestinal neoplasia (Min) mouse mutation causes adenomas to develop throughout the small and large intestines. The BALB Min (C.B6-Apc(Min/+)) congenic strain was generated by backcrossing into BALB/c the Apc(Min) allele from C57BL/6J-Apc(Min/+) mice. BALB-Min mice have a low tumor multiplicity (27.4 small intestine tumors/mouse) and a relatively long life span (>1 year) that makes them amenable to long-term studies. To investigate the interplay of the adaptive immune system and intestinal tumorigenesis, the immunodeficient compound mutant strain BALB-RagMin (C.Cg-Rag2(-/-) Apc(Min/+)) was generated. BALB-RagMin mice had a significant increase in tumors in the small, but not large, intestine relative to their BALB-Min counterparts (43.0 versus 24.0 tumors/mouse, respectively). The results suggest that the adaptive immune system plays a role in either the elimination or the equilibrium phase of cancer immunoediting in the small intestine in this model. We investigated the effect of the enterohepatic bacterial pathogen Helicobacter hepaticus on liver and intestine tumorigenesis in BALB-RagMin mice. H. hepaticus-infected BALB RagMin mice developed moderate hepatitis, moderate typhlitis, and mild colitis. There were no differences in small intestine and cecal tumor multiplicity, regionality, or size relative to that in uninfected mice. However, H. hepaticus infected BALB-RagMin mice had a significant increase in colon tumor incidence relative to uninfected BALB-RagMin mice (23.5% versus 1.7%, respectively). The data suggest that H. hepaticus, which is present in many research colonies, promotes colon tumorigenesis in the BALB-RagMin mouse and that it has the potential to confound colon tumorigenesis studies. PMID- 18411293 TI - The signal recognition particle pathway is required for virulence in Streptococcus pyogenes. AB - The signal recognition particle (SRP) pathway is a universally conserved pathway for targeting polypeptides for secretion via the cotranslational pathway. In particular, the SRP pathway is thought to be the main mechanism for targeting polypeptides in gram-positive bacteria, including a number of important human pathogens. Though widely considered to be an essential cellular component, recent advances have indicated this pathway may be dispensable in gram-positive bacteria of the genus Streptococcus under in vitro conditions. However, its importance for the pathogenesis of streptococcal disease is unknown. In this study, we investigated the importance of the SRP pathway for virulence factor secretion in the human pathogen Streptococcus pyogenes. While the SRP pathway was not found to be essential for viability in vitro, SRP mutants demonstrated a medium-specific growth defect that could be rescued by the addition of glucose. We also observed that a distinct subset of virulence factors were dependent upon the SRP pathway for secretion, whereas others were completely independent of this pathway. Significantly, deletion of the SRP pathway resulted in mutants that were highly attenuated in both a zebrafish model of necrotic myositis and a murine subcutaneous ulcer model, highlighting the importance of this pathway in vivo. These studies emphasize the importance of the SRP pathway for the in vivo survival and pathogenesis of S. pyogenes. PMID- 18411294 TI - Initial delay in the immune response to Francisella tularensis is followed by hypercytokinemia characteristic of severe sepsis and correlating with upregulation and release of damage-associated molecular patterns. AB - "Francisella tularensis subsp. novicida" intranasal infection causes a rapid pneumonia in mice with mortality at 4 to 6 days with a low dose of bacteria (10(2) bacteria). The short time to death suggests that there is a failure of the innate immune response. As the neutrophil is often the first cell type to infiltrate sites of infection, we focused on the emigration of neutrophils in this infection, as well as cytokines involved in their recruitment. The results indicated that there was a significant delay in the influx of neutrophils into the bronchoalveolar lavage fluid of F. tularensis subsp. novicida-infected mice. The delay in neutrophil recruitment in F. tularensis subsp. novicida-infected mice correlated with a delay in the upregulation of multiple proinflammatory cytokines and chemokines, as well as a delay in caspase-1 activation. Strikingly, the initial delay in the upregulation of cytokines through 1 day postinfection was followed by profound upregulation of multiple cytokines and chemokines to levels consistent with hypercytokinemia described for severe sepsis. This finding was further supported by a bacteremia and the cellular relocalization and release of high-mobility group box-1 and S100A9, both of which are damage-associated molecular pattern molecules and are known to be mediators of severe sepsis. PMID- 18411295 TI - Fas/CD95-mediated apoptosis of type II cells is blocked by Toxoplasma gondii primarily via interference with the mitochondrial amplification loop. AB - The intracellular protozoan Toxoplasma gondii induces persistent infections in various hosts and is an important opportunistic pathogen of humans with immature or deficient immune responses. The ability to survive intracellularly largely depends on the blocking of different proapoptotic signaling cascades of its host cell. Fas/CD95 triggers an apoptotic cascade that is crucial for immunity and the outcome of infectious diseases. We have determined the mechanism by which T. gondii counteracts death receptor-mediated cell death in type II cells that transduce Fas/CD95 ligation via caspase 8-mediated activation of the mitochondrial amplification loop. The results showed that infection with T. gondii significantly reduced Fas/CD95-triggered apoptosis in HeLa cells by inhibiting the activities of initiator caspases 8 and 9 and effector caspase 3/7. Parasitic infection dose dependently diminished cleavage of caspase 8, the BH3 only protein Bid, and the downstream caspases 9 and 3. Importantly, interference with Fas/CD95-triggered caspase 8 and caspase 3/7 activities after parasitic infection was largely dependent on the presence of caspase 9. Within the mitochondrial amplification loop, T. gondii significantly inhibited the Fas/CD95 triggered release of cytochrome c into the host cell cytosol. These results indicate that T. gondii inhibits Fas/CD95-mediated apoptosis in type II cells primarily by decreasing the apoptogenic function of mitochondria. PMID- 18411296 TI - The surface-exposed carboxyl region of Mycoplasma pneumoniae elongation factor Tu interacts with fibronectin. AB - Mycoplasma pneumoniae is a bacterial pathogen of the human respiratory tract that causes a wide range of airway diseases as well as extrapulmonary symptoms. It possesses a distinct, differentiated terminal structure, termed the attachment organelle, that mediates adherence to the host respiratory epithelium. Previously, we reported that surface-associated M. pneumoniae elongation factor Tu (EF-Tu, also called MPN665) serves as a fibronectin (Fn)-binding protein, facilitating interactions between mycoplasmas and extracellular matrix. In the present study, we determined that binding of M. pneumoniae EF-Tu to Fn is primarily mediated by the EF-Tu carboxyl region. A 179-amino-acid region spanning the carboxyl terminus (designated EC; amino acids 192 to 394) binds Fn in a dose dependent manner. Further analysis of carboxyl constructs (ED3 and ED4) and their deletion truncations (ED3.1, ED3.2, and ED4.1) revealed that the carboxyl region possessed two distinct sites with different Fn-binding efficiencies. Immunogold electron microscopy using antibodies raised against recombinant ED3 and ED4 demonstrated the surface accessibility of the EF-Tu carboxyl region. Competitive binding assays using intact radiolabeled mycoplasmas and purified recombinant ED3 and ED4 proteins, along with antibody blocking assays, reinforced the role of the surface-exposed EF-Tu carboxyl region in Fn binding. Alkali and high-salt treatment of mycoplasma membranes and Triton X-114-partitioned mycoplasma fractions confirmed the stable association of EF-Tu within the mycoplasma membrane. These observations highlight the unique, multifaceted, and unpredictable role of the classically defined cytoplasmic protein EF-Tu relative to cellular function, compartmentalization, and topography. PMID- 18411297 TI - DEC1 modulates the circadian phase of clock gene expression. AB - DEC1 suppresses CLOCK/BMAL1-enhanced promoter activity, but its role in the circadian system of mammals remains unclear. Here we examined the effect of Dec1 overexpression or deficiency on circadian gene expression triggered with 50% serum. Overexpression of Dec1 delayed the phase of clock genes such as Dec1, Dec2, Per1, and Dbp that contain E boxes in their regulatory regions, whereas it had little effect on the circadian phase of Per2 and Cry1 carrying CACGTT E' boxes. In contrast, Dec1 deficiency advanced the phase of the E-box-containing clock genes but not that of the E'-box-containing clock genes. Accordingly, DEC1 showed strong binding and transrepression on the E box, but not on the E' box, in chromatin immunoprecipitation, electrophoretic mobility shift, and luciferase reporter assays. Dec1-/- mice showed behavioral rhythms with slightly but significantly longer circadian periods under conditions of constant darkness and faster reentrainment to a 6-h phase-advanced shift of a light-dark cycle. Knockdown of Dec2 with small interfering RNA advanced the phase of Dec1 and Dbp expression, and double knockdown of Dec1 and Dec2 had much stronger effects on the expression of the E-box-containing clock genes. These findings suggest that DEC1, along with DEC2, plays a role in the finer regulation and robustness of the molecular clock. PMID- 18411298 TI - CHIP deficiency decreases longevity, with accelerated aging phenotypes accompanied by altered protein quality control. AB - During the course of biological aging, there is a gradual accumulation of damaged proteins and a concomitant functional decline in the protein degradation system. Protein quality control is normally ensured by the coordinated actions of molecular chaperones and the protein degradation system that collectively help to maintain protein homeostasis. The carboxyl terminus of Hsp70-interacting protein (CHIP), a ubiquitin ligase/cochaperone, participates in protein quality control by targeting a broad range of chaperone substrates for proteasome degradation via the ubiquitin-proteasome system, demonstrating a broad involvement of CHIP in maintaining cytoplasmic protein quality control. In the present study, we have investigated the influence that protein quality control exerts on the aging process by using CHIP-/- mice. CHIP deficiency in mice leads to a markedly reduced life span, along with accelerated age-related pathophysiological phenotypes. These features were accompanied by indications of accelerated cellular senescence and increased indices of oxidative stress. In addition, CHIP /- mice exhibit a deregulation of protein quality control, as indicated by elevated levels of toxic oligomer proteins and a decline in proteasome activity. Taken together, these data reveal that impaired protein quality control contributes to cellular senescence and implicates CHIP-dependent quality control mechanisms in the regulation of mammalian longevity in vivo. PMID- 18411299 TI - Ribonomic analysis of human Pum1 reveals cis-trans conservation across species despite evolution of diverse mRNA target sets. AB - PUF family proteins are among the best-characterized regulatory RNA-binding proteins in nonmammalian species, but relatively little is known about mRNA targets or functions of mammalian PUF proteins. In this study, we used ribonomic analysis to identify and analyze mRNAs associated with ribonucleoproteins containing an endogenous human PUF protein, Pum1. Pum1-associated mRNAs were highly enriched for genes encoding proteins that function in transcriptional regulation and cell cycle/proliferation, results consistent with the posttranscriptional RNA regulon model and the proposed ancestral functions of PUF proteins in stem cell biology. Analysis of 3' untranslated region sequences of Pum1-associated mRNAs revealed a core Pum1 consensus sequence, UGUAHAUA. Pum1 knockdown demonstrated that Pum1 enhances decay of associated mRNAs, and relocalization of Pum1 to stress granules suggested that Pum1 functions in repression of translation. This study is the first in vivo genome-wide mRNA target identification of a mammalian PUF protein and provides direct evidence that human PUF proteins regulate stability of associated mRNAs. Comparison of Pum1-associated mRNAs to mRNA targets of PUF proteins from Saccharomyces cerevisiae and Drosophila melanogaster demonstrates how a well-conserved RNA binding domain and cognate binding sequence have been evolutionarily rewired to regulate the collective expression of different sets of functionally related genes. PMID- 18411300 TI - Identification of proteins associating with glycosylphosphatidylinositol- anchored T-cadherin on the surface of vascular endothelial cells: role for Grp78/BiP in T-cadherin-dependent cell survival. AB - There is scant knowledge regarding how cell surface lipid-anchored T-cadherin (T cad) transmits signals through the plasma membrane to its intracellular targets. This study aimed to identify membrane proteins colocalizing with atypical glycosylphosphatidylinositol (GPI)-anchored T-cad on the surface of endothelial cells and to evaluate their role as signaling adaptors for T-cad. Application of coimmunoprecipitation from endothelial cells expressing c-myc-tagged T-cad and high-performance liquid chromatography revealed putative association of T-cad with the following proteins: glucose-related protein GRP78, GABA-A receptor alpha1 subunit, integrin beta3, and two hypothetical proteins, LOC124245 and FLJ32070. Association of Grp78 and integrin beta3 with T-cad on the cell surface was confirmed by surface biotinylation and reciprocal immunoprecipitation and by confocal microscopy. Use of anti-Grp78 blocking antibodies, Grp78 small interfering RNA, and coexpression of constitutively active Akt demonstrated an essential role for surface Grp78 in T-cad-dependent survival signal transduction via Akt in endothelial cells. The findings herein are relevant in the context of both the identification of transmembrane signaling partners for GPI-anchored T cad as well as the demonstration of a novel mechanism whereby Grp78 can influence endothelial cell survival as a cell surface signaling receptor rather than an intracellular chaperone. PMID- 18411301 TI - The TSC1-TSC2 complex is required for proper activation of mTOR complex 2. AB - The mammalian target of rapamycin (mTOR) is a protein kinase that forms two functionally distinct complexes important for nutrient and growth factor signaling. Both complexes phosphorylate a hydrophobic motif on downstream protein kinases, which contributes to the activation of these kinases. mTOR complex 1 (mTORC1) phosphorylates S6K1, while mTORC2 phosphorylates Akt. The TSC1-TSC2 complex is a critical negative regulator of mTORC1. However, how mTORC2 is regulated and whether the TSC1-TSC2 complex is involved are unknown. We find that mTORC2 isolated from a variety of cells lacking a functional TSC1-TSC2 complex is impaired in its kinase activity toward Akt. Importantly, the defect in mTORC2 activity in these cells can be separated from effects on mTORC1 signaling and known feedback mechanisms affecting insulin receptor substrate-1 and phosphatidylinositol 3-kinase. Our data also suggest that the TSC1-TSC2 complex positively regulates mTORC2 in a manner independent of its GTPase-activating protein activity toward Rheb. Finally, we find that the TSC1-TSC2 complex can physically associate with mTORC2 but not mTORC1. These data demonstrate that the TSC1-TSC2 complex inhibits mTORC1 and activates mTORC2, which through different mechanisms promotes Akt activation. PMID- 18411302 TI - Multiple yeast genes, including Paf1 complex genes, affect telomere length via telomerase RNA abundance. AB - Twofold reductions in telomerase RNA levels cause telomere shortening in both humans and the yeast Saccharomyces cerevisiae. To test whether multiple genes that affect telomere length act by modulating telomerase RNA abundance, we used real-time reverse transcription-PCR to screen S. cerevisiae deletion strains reported to maintain shorter or longer telomeres to determine the levels of their telomerase RNA (TLC1) abundance. Of 290 strains screened, 5 had increased TLC1 levels; 4 of these maintained longer telomeres. Twenty strains had decreased TLC1 levels; 18 of these are known to maintain shorter telomeres. Four strains with decreased TLC1 RNA levels contained deletions of subunits of Paf1C (polymerase II associated factor complex). While Paf1C had been implicated in the transcription of both polyadenylated and nonpolyadenylated RNAs, Paf1C had not been associated previously with the noncoding telomerase RNA. In Paf1C mutant strains, TLC1 overexpression partially rescues telomere length and cell growth defects, suggesting that telomerase RNA is a critical direct or indirect Paf1C target. Other factors newly identified as affecting TLC1 RNA levels include cyclin dependent kinase, the mediator complex, protein phosphatase 2A, and ribosomal proteins L13B and S16A. This report establishes that a subset of telomere length genes act by modulating telomerase RNA abundance. PMID- 18411303 TI - Phosphorylation of fibroblast growth factor (FGF) receptor 1 at Ser777 by p38 mitogen-activated protein kinase regulates translocation of exogenous FGF1 to the cytosol and nucleus. AB - Exogenous fibroblast growth factor 1 (FGF1) signals through activation of transmembrane FGF receptors (FGFRs) but may also regulate cellular processes after translocation to the cytosol and nucleus of target cells. Translocation of FGF1 occurs across the limiting membrane of intracellular vesicles and is a regulated process that depends on the C-terminal tail of the FGFR. Here, we report that translocation of FGF1 requires activity of the alpha isoform of p38 mitogen-activated protein kinase (MAPK). FGF1 translocation was inhibited after chemical inhibition of p38 MAPK or after small interfering RNA knockdown of p38alpha. Translocation was increased after stimulation of p38 MAPK with anisomycin, mannitol, or H2O2. The activity level of p38 MAPK was not found to affect endocytosis or intracellular sorting of FGF1/FGFR1. Instead, we found that p38 MAPK regulates FGF1 translocation by phosphorylation of FGFR1 at Ser777. The FGFR1 mutation S777A abolished FGF1 translocation, while phospho-mimetic mutations of Ser777 to Asp or Glu allowed translocation to take place and bypassed the requirement for active p38 MAPK. Ser777 in FGFR1 was directly phosphorylated by p38alpha in a cell-free system. These data demonstrate a crucial role for p38alpha MAPK in the regulated translocation of exogenous FGF1 into the cytosol/nucleus, and they reveal a specific role for p38alpha MAPK mediated serine phosphorylation of FGFR1. PMID- 18411304 TI - Pak1 and Pak2 mediate tumor cell invasion through distinct signaling mechanisms. AB - Pak kinases are thought to play critical roles in cell migration and invasion. Here, we analyze the roles of Pak1 and Pak2 in breast carcinoma cell invasion using the transient transfection of small interfering RNA. We find that although both Pak1 and Pak2 contribute to breast carcinoma invasion stimulated by heregulin, these roles are mediated by distinct signaling mechanisms. Thus, whereas the depletion of Pak1 interferes with the heregulin-mediated dephosphorylation of cofilin, the depletion of Pak2 does not. The depletion of Pak1 also has a stronger inhibitory effect on lamellipodial protrusion than does the depletion of Pak2. Interestingly, Pak1 and Pak2 play opposite roles in regulating the phosphorylation of the myosin light chain (MLC). Whereas the depletion of Pak1 decreases phospho-MLC levels in heregulin-stimulated cells, the depletion of Pak2 enhances MLC phosphorylation. Consistent with their opposite effects on MLC phosphorylation, Pak1 and Pak2 differentially modulate focal adhesions. Pak2-depleted cells display an increase in focal adhesion size, whereas in Pak1-depleted cells, focal adhesions fail to mature. We also found that the depletion of Pak2, but not Pak1, enhances RhoA activity and that the inhibition of RhoA signaling in Pak2-depleted cells decreases MLC phosphorylation and restores cell invasion. In summary, this work presents the first comprehensive analysis of functional differences between the Pak1 and Pak2 isoforms. PMID- 18411305 TI - Emilin1 deficiency causes structural and functional defects of lymphatic vasculature. AB - Lymphatic-vasculature function critically depends on extracellular matrix (ECM) and on its connections with lymphatic endothelial cells (LECs). However, the composition and the architecture of ECM have not been fully taken into consideration in studying the biology and the pathology of the lymphatic system. EMILIN1, an elastic microfibril-associated protein, is highly expressed by LECs in vitro and colocalizes with lymphatic vessels in several mouse tissues. A comparative study between WT and Emilin1-/- mice highlighted the fact that Emilin1 deficiency in both CD1 and C57BL/6 backgrounds results in hyperplasia, enlargement, and frequently an irregular pattern of superficial and visceral lymphatic vessels and in a significant reduction of anchoring filaments. Emilin1 deficient mice also develop larger lymphangiomas than WT mice. Lymphatic vascular morphological alterations are accompanied by functional defects, such as mild lymphedema, a highly significant drop in lymph drainage, and enhanced lymph leakage. Our findings demonstrate that EMILIN1 is involved in the regulation of the growth and in the maintenance of the integrity of lymphatic vessels, a fundamental requirement for efficient function. The phenotype displayed by Emilin1(-/-) mice is the first abnormal lymphatic phenotype associated with the deficiency of an ECM protein and identifies EMILIN1 as a novel local regulator of lymphangiogenesis. PMID- 18411306 TI - Leptin affects endocardial cushion formation by modulating EMT and migration via Akt signaling cascades. AB - Blood circulation is dependent on heart valves to direct blood flow through the heart and great vessels. Valve development relies on epithelial to mesenchymal transition (EMT), a central feature of embryonic development and metastatic cancer. Abnormal EMT and remodeling contribute to the etiology of several congenital heart defects. Leptin and its receptor were detected in the mouse embryonic heart. Using an ex vivo model of cardiac EMT, the inhibition of leptin results in a signal transducer and activator of transcription 3 and Snail/vascular endothelial cadherin-independent decrease in EMT and migration. Our data suggest that an Akt signaling pathway underlies the observed phenotype. Furthermore, loss of leptin phenocopied the functional inhibition of alphavbeta3 integrin receptor and resulted in decreased alphavbeta3 integrin and matrix metalloprotease 2, suggesting that the leptin signaling pathway is involved in adhesion and migration processes. This study adds leptin to the repertoire of factors that mediate EMT and, for the first time, demonstrates a role for the interleukin 6 family in embryonic EMT. PMID- 18411307 TI - Phosphorylation of SDT repeats in the MDC1 N terminus triggers retention of NBS1 at the DNA damage-modified chromatin. AB - DNA double-strand breaks (DSBs) trigger accumulation of the MRE11-RAD50-Nijmegen breakage syndrome 1 (NBS1 [MRN]) complex, whose retention on the DSB-flanking chromatin facilitates survival. Chromatin retention of MRN requires the MDC1 adaptor protein, but the mechanism behind the MRN-MDC1 interaction is unknown. We show that the NBS1 subunit of MRN interacts with the MDC1 N terminus enriched in Ser-Asp-Thr (SDT) repeats. This interaction was constitutive and mediated by binding between the phosphorylated SDT repeats of MDC1 and the phosphate-binding forkhead-associated domain of NBS1. Phosphorylation of the SDT repeats by casein kinase 2 (CK2) was sufficient to trigger MDC1-NBS1 interaction in vitro, and MDC1 associated with CK2 activity in cells. Inhibition of CK2 reduced SDT phosphorylation in vivo, and disruption of the SDT-associated phosphoacceptor sites prevented the retention of NBS1 at DSBs. Together, these data suggest that phosphorylation of the SDT repeats in the MDC1 N terminus functions to recruit NBS1 and, thereby, increases the local concentration of MRN at the sites of chromosomal breakage. PMID- 18411308 TI - Constitutive phosphorylation of MDC1 physically links the MRE11-RAD50-NBS1 complex to damaged chromatin. AB - The MRE11-RAD50-Nijmegen breakage syndrome 1 (NBS1 [MRN]) complex accumulates at sites of DNA double-strand breaks (DSBs) in microscopically discernible nuclear foci. Focus formation by the MRN complex is dependent on MDC1, a large nuclear protein that directly interacts with phosphorylated H2AX. In this study, we identified a region in MDC1 that is essential for the focal accumulation of the MRN complex at sites of DNA damage. This region contains multiple conserved acidic sequence motifs that are constitutively phosphorylated in vivo. We show that these motifs are efficiently phosphorylated by caseine kinase 2 (CK2) in vitro and directly interact with the N-terminal forkhead-associated domain of NBS1 in a phosphorylation-dependent manner. Mutation of these conserved motifs in MDC1 or depletion of CK2 by small interfering RNA disrupts the interaction between MDC1 and NBS1 and abrogates accumulation of the MRN complex at sites of DNA DSBs in vivo. Thus, our data reveal the mechanism by which MDC1 physically couples the MRN complex to damaged chromatin. PMID- 18411309 TI - DDA3 recruits microtubule depolymerase Kif2a to spindle poles and controls spindle dynamics and mitotic chromosome movement. AB - Dynamic turnover of the spindle is a driving force for chromosome congression and segregation in mitosis. Through a functional genomic analysis, we identify DDA3 as a previously unknown regulator of spindle dynamics that is essential for mitotic progression. DDA3 depletion results in a high frequency of unaligned chromosomes, a substantial reduction in tension across sister kinetochores at metaphase, and a decrease in the velocity of chromosome segregation at anaphase. DDA3 associates with the mitotic spindle and controls microtubule (MT) dynamics. Mechanistically, DDA3 interacts with the MT depolymerase Kif2a in an MT-dependent manner and recruits Kif2a to the mitotic spindle and spindle poles. Depletion of DDA3 increases the steady-state levels of spindle MTs by reducing the turnover rate of the mitotic spindle and by increasing the rate of MT polymerization, which phenocopies the effects of partial knockdown of Kif2a. Thus, DDA3 represents a new class of MT-destabilizing protein that controls spindle dynamics and mitotic progression by regulating MT depolymerases. PMID- 18411310 TI - Flightless-I regulates proinflammatory caspases by selectively modulating intracellular localization and caspase activity. AB - Caspase-1 and caspase-11 are proinflammatory caspases that regulate cytokine production and leukocyte migration during pathogen infection. In an attempt to identify new intracellular regulators of caspase-11, we found that Flightless-I, a member of the gelsolin superfamily of actin-remodeling proteins, interacts and regulates both caspase-11 and caspase-1. Flightless-I targets caspase-11 to the Triton X-100-insoluble cytoskeleton fraction and the cell leading edge. In addition, Flightless-I inhibits caspase-1 activation and caspase-1-mediated interleukine-1beta (IL-1beta) maturation. The physiological relevance of these findings is supported by the opposing effects of Flightless-I overexpression and knockdown on caspase-1 activity and IL-1beta maturation. Our results suggest that Flightless-I may be a bona fide caspase-1 inhibitor that acts through a mechanism similar to that of cytokine response modifier A, a potent caspase-1 inhibitor from the cowpox virus. Our study provides a new mechanism controlling the localization and activation of proinflammatory caspases. PMID- 18411311 TI - Cell polarization during monopolar cytokinesis. AB - During cytokinesis, a specialized set of proteins is recruited to the equatorial region between spindle poles by microtubules and actin filaments, enabling furrow assembly and ingression before cell division. We investigate the mechanisms underlying regional specialization of the cytoskeleton in HeLa cells undergoing drug-synchronized monopolar cytokinesis. After forced mitotic exit, the cytoskeleton of monopolar mitotic cells is initially radially symmetric but undergoes a symmetry-breaking reaction that simultaneously polarizes microtubules and the cell cortex, with a concentration of cortical furrow markers into a cap at one side of the cell. Polarization requires microtubules, F-actin, RhoA, Myosin II activity, and Aurora B kinase activity. Aurora B localizes to actin cables in a gap between the monopolar midzone and the furrow-like cortex, suggesting a communication between them. We propose that feedback loops between cortical furrow components and microtubules promote symmetry breaking during monopolar cytokinesis and regional specialization of the cytoskeleton during normal bipolar cytokinesis. PMID- 18411312 TI - Spinophilin participates in information transfer at immunological synapses. AB - The adaptive immune response is initiated by the presentation of peptides bound to major histocompatibility complex molecules on dendritic cells (DCs) to antigen specific T lymphocytes at a junction termed the immunological synapse. Although much attention has been paid to cytoplasmic events on the T cell side of the synapse, little is known concerning events on the DC side. We have sought signal transduction components of the neuronal synapse that were also expressed by DCs. One such protein is spinophilin, a scaffolding protein of neuronal dendritic spines that regulates synaptic transmission. In inactive, immature DCs, spinophilin is located throughout the cytoplasm but redistributes to the plasma membrane upon stimulus-induced maturation. In DCs interacting with T cells, spinophilin is polarized dynamically to contact sites in an antigen-dependent manner. It is also required for optimal T cell activation because DCs derived from mice lacking spinophilin exhibit defects in antigen presentation both in vitro and in vivo. Thus, spinophilin may play analogous roles in information transfer at both neuronal and immunological synapses. PMID- 18411313 TI - Decoding ARE-mediated decay: is microRNA part of the equation? AB - Messenger ribonucleic acids (mRNAs) containing adenine/uridine-rich elements (AREs) in their 3' untranslated region are particularly labile, allowing for the regulation of expression for growth factors, oncoproteins, and cytokines. The regulators, effectors, and location of ARE-mediated decay (AMD) have been investigated by many groups in recent years, and several links have been found between AMD and microRNA-mediated decay. We highlight these similarities, along with recent advances in the field of AMD, and also mention how there is still much left unknown surrounding this specialized mode of mRNA decay. PMID- 18411314 TI - Spinophilin and the immune synapse. AB - Extensive alterations in cellular organization are known to accompany the responses of sensitized T cells to target cells presenting an antigen of interest. Now, equally if not more dramatic changes are found to take place in cells presenting an antigen. With the help of a spinophilin-GFP fusion protein, Bloom et al. (Bloom, O., J.J. Unternaehrer, A. Jiang, J.-S. Shin, L. Delamarre, P. Allen, and I. Mellman. 2008. J. Cell Biol. 181:203-211) have captured a remarkable polarization of the cellular architecture of dendritic cells presenting an antigen to T cells. PMID- 18411316 TI - Antimicrobial susceptibilities of geographically diverse clinical human isolates of Leptospira. AB - Although antimicrobial therapy of leptospirosis has been studied in a few randomized controlled clinical studies, those studies were limited to specific regions of the world and few have characterized infecting strains. A broth microdilution technique for the assessment of antibiotic susceptibility has been developed at Brooke Army Medical Center. In the present study, we assessed the susceptibilities of 13 Leptospira isolates (including recent clinical isolates) from Egypt, Thailand, Nicaragua, and Hawaii to 13 antimicrobial agents. Ampicillin, cefepime, azithromycin, and clarithromycin were found to have MICs below the lower limit of detection (0.016 microg/ml). Cefotaxime, ceftriaxone, imipenem-cilastatin, penicillin G, moxifloxacin, ciprofloxacin, and levofloxacin had MIC(90)s between 0.030 and 0.125 microg/ml. Doxycycline and tetracycline had the highest MIC(90)s: 2 and 4 microg/ml, respectively. Doxycycline and tetracycline were noted to have slightly higher MICs against isolates from Egypt than against strains from Thailand or Hawaii; otherwise, the susceptibility patterns were similar. There appears to be possible variability in susceptibility to some antimicrobial agents among strains, suggesting that more extensive testing to look for geographic variability should be pursued. PMID- 18411315 TI - Whole-genome pyrosequencing of an epidemic multidrug-resistant Acinetobacter baumannii strain belonging to the European clone II group. AB - The whole-genome sequence of an epidemic, multidrug-resistant Acinetobacter baumannii strain (strain ACICU) belonging to the European clone II group and carrying the plasmid-mediated bla(OXA)(-)(58) carbapenem resistance gene was determined. The A. baumannii ACICU genome was compared with the genomes of A. baumannii ATCC 17978 and Acinetobacter baylyi ADP1, with the aim of identifying novel genes related to virulence and drug resistance. A. baumannii ACICU has a single chromosome of 3,904,116 bp (which is predicted to contain 3,758 genes) and two plasmids, pACICU1 and pACICU2, of 28,279 and 64,366 bp, respectively. Genome comparison showed 86.4% synteny with A. baumannii ATCC 17978 and 14.8% synteny with A. baylyi ADP1. A conspicuous number of transporters belonging to different superfamilies was predicted for A. baumannii ACICU. The relative number of transporters was much higher in ACICU than in ATCC 17978 and ADP1 (76.2, 57.2, and 62.5 transporters per Mb of genome, respectively). An antibiotic resistance island, AbaR2, was identified in ACICU and had plausibly evolved by reductive evolution from the AbaR1 island previously described in multiresistant strain A. baumannii AYE. Moreover, 36 putative alien islands (pAs) were detected in the ACICU genome; 24 of these had previously been described in the ATCC 17978 genome, 4 are proposed here for the first time and are present in both ATCC 17978 and ACICU, and 8 are unique to the ACICU genome. Fifteen of the pAs in the ACICU genome encode genes related to drug resistance, including membrane transporters and ex novo acquired resistance genes. These findings provide novel insight into the genetic basis of A. baumannii resistance. PMID- 18411317 TI - Susceptibility of Chlamydia trachomatis to the excipient hydroxyethyl cellulose: pH and concentration dependence of antimicrobial activity. AB - Hydroxyethyl cellulose (HEC) is used as a neutral excipient in microbicides used against sexually transmitted pathogens. However, HEC inhibits the infection of cervical epithelial cells by Chlamydia trachomatis at pH 5 in a concentration dependent manner. At pH 7, infection is inversely dependent on the concentration of HEC, possibly due to pH-dependent calcium sequestration. PMID- 18411318 TI - In vitro susceptibility of isolates of Francisella tularensis types A and B from North America. AB - Due to concern that Francisella tularensis, the causative agent of tularemia, may be used as a bioterrorist weapon, the Clinical and Laboratory Standards Institute recently provided a susceptibility testing method with breakpoints. Here, 169 isolates (92 type A and 77 type B) from North America were tested against seven antimicrobial agents (streptomycin, gentamicin, tetracycline, doxycycline, ciprofloxacin, levofloxacin, and chloramphenicol) used for the treatment of tularemia. The MICs for all of the isolates fell within the susceptible range. In addition, all isolates had MICs for erythromycin of 0.5 to 4 microg/ml, in contrast to an MIC of >256 microg/ml for the common laboratory strain LVS (live vaccine strain). PMID- 18411319 TI - First case of emergence of atovaquone-proguanil resistance in Plasmodium falciparum during treatment in a traveler in Comoros. PMID- 18411320 TI - Local treatment of experimental Pseudomonas aeruginosa osteomyelitis with a biodegradable dilactide polymer releasing ciprofloxacin. AB - A biodegradable system of poly-D,L-dilactide releasing ciprofloxacin was assessed in a Pseudomonas aeruginosa osteomyelitis model after inoculation of the test pathogen into the left tibia of 76 New Zealand White rabbits; 31 were controls (group A), and 45 were implanted with the polymer at the infection site (group B). The rabbits were killed on a weekly basis, and cancellous bone was harvested for histopathology and for estimation of bacterial growth and the concentrations of ciprofloxacin. Tibial X ray was performed immediately before the animals were killed. The total number of fistulas with purulent discharge that developed after inoculation of the pathogen was counted, and fistulas with purulent discharge were found in 16 animals in group A (51.6%) and 3 animals in group B (6.7%) (P < 0.0001). The animals in group A had a profound loss of body weight compared to the animals in group B. The main radiological finding was the presence of sequestra in 25 animals (80.6%) in group A and 6 animals in group B (13.3%) (P < 0.0001). The bacterial load in group B was significantly reduced compared to that in group A, possibly due to the prolonged local antibiotic release at concentrations exceeding even 80 times the MIC for the test pathogen. The histology of animals killed after week 49 revealed a mild inflammatory reaction accompanied by diffuse fibrosis and new bone formation in group A animals and the presence of small polymer particles in group B animals. It is concluded that the system described achieved eradication of the pathogen, accompanied by clinical and radiologically confirmed benefits, so this treatment may be a candidate for the management of difficult orthopedic infections. PMID- 18411321 TI - Activities of clindamycin, daptomycin, doxycycline, linezolid, trimethoprim sulfamethoxazole, and vancomycin against community-associated methicillin resistant Staphylococcus aureus with inducible clindamycin resistance in murine thigh infection and in vitro pharmacodynamic models. AB - Controversy exists about the most effective treatment options for community associated methicillin-resistant Staphylococcus aureus (CA-MRSA) and about the ability of these strains to develop inducible resistance to clindamycin during therapy. Using both in vitro pharmacodynamic and murine thigh infection models, we evaluated and compared several antimicrobial compounds against CA-MRSA. Strains with inducible macrolide lincosamide-streptogramin type B (iMLS(B)) resistance and strains in which resistance was noninducible were evaluated. Two levels of inocula (10(5) and 10(7)) were evaluated for clindamycin activity in the in vivo model. In both models, the antimicrobial evaluation was performed in triplicate, and bacterial quantification occurred over 72 h, with drug doses that were designed to simulate the free drug area-under-the-concentration-time curve values (fAUCs) obtained from human samples. When the activity of clindamycin against the iMLS(B) strains was evaluated, constitutive resistance was noted at 24 h (MIC of >256), and failure was noted at an inoculum of > or =10(6) in the in vivo models. However, at a low inoculum (10(5)) in the murine thigh-infection model, clindamycin demonstrated modest activity, reducing the CFU/thigh count for clindamycin resistance-inducible strains at 72 h (0.45 to 1.3 logs). Overall, administration of daptomycin followed by vancomycin demonstrated the most significant kill against all strains in both models. Against the clindamycin noninducible strain, clindamycin and doxycycline demonstrated significant kill. Doxycycline, linezolid, and trimethoprim-sulfamethoxazide (not run in the murine model) demonstrated bacteriostatic activity against clindamycin resistance inducible isolates. This study demonstrates that clindamycin's activity against the iMLS(B) strains tested is partially impacted by inoculum size. At present, there are several alternatives that appear promising for treating clindamycin resistance-inducible strains of CA-MRSA. PMID- 18411322 TI - Pharmacodynamic characterization of ceftobiprole in experimental pneumonia caused by phenotypically diverse Staphylococcus aureus strains. AB - Ceftobiprole (BPR) is an investigational cephalosporin with activity against Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA) strains. The pharmacodynamic (PD) profile of BPR against S. aureus strains with a variety of susceptibility phenotypes in an immunocompromised murine pneumonia model was characterized. The BPR MICs of the test isolates ranged from 0.25 to 2 mug/ml. Pharmacokinetic (PK) studies were conducted with infected neutropenic BALB/c mice; and the BPR concentrations were measured in plasma, epithelial lining fluid (ELF), and lung tissue. PD studies with these mice were undertaken with eight S. aureus isolates (two methicillin-susceptible S. aureus strains, three hospital acquired MRSA strains, and three community-acquired MRSA strains). Subcutaneous BPR doses of 2 to 125 mg/kg of body weight/day were administered, and the change in the number of log(10) CFU/ml in lungs was evaluated after 24 h of therapy. The PD profile was characterized by using the free drug exposures (f) determined from the following parameters: the percentage of time that the concentration was greater than the MIC (T > MIC), the maximum concentration in serum/MIC, and the area under the concentration-time curve/MIC. The BPR PK parameters were linear over the dose range studied in plasma, and the ELF concentrations ranged from 60 to 94% of the free plasma concentration. fT > MIC was the parameter that best correlated with efficacy against a diverse array of S. aureus isolates in this murine pneumonia model. The 80% effective dose (ED(80)), ED(50), and stasis exposures appeared to be similar among the isolates studied. BPR exerted maximal antibacterial effects when fT > MIC ranged from 6 to 22%, regardless of the phenotypic profile of resistance to beta-lactam, fluoroquinolone, erythromycin, clindamycin, or tetracycline antibiotics. PMID- 18411323 TI - Decrease of atazanavir and lopinavir plasma concentrations in a boosted double human immunodeficiency virus protease inhibitor salvage regimen. AB - The human immunodeficiency virus protease inhibitor combination of atazanavir (ATV)-lopinavir-ritonavir was reported to exhibit a mutual pharmacoenhancement of plasma lopinavir and ATV concentrations which may be beneficial for salvage patients. We identified 17 patients in our pharmacokinetic database taking this combination and found conflicting results. Plasma concentrations of both ATV and lopinavir were modestly, although not significantly, decreased when the drugs were coadministered. Therefore, patients should be selected carefully for this regimen and frequent clinical and therapeutic drug monitoring is strongly advised. PMID- 18411324 TI - Selection of replicon variants resistant to ACH-806, a novel hepatitis C virus inhibitor with no cross-resistance to NS3 protease and NS5B polymerase inhibitors. AB - We have discovered a novel class of compounds active against hepatitis C virus (HCV), using a surrogate cellular system, HCV replicon cells. The leading compound in the series, ACH-806 (GS-9132), is a potent and specific inhibitor of HCV. The selection of resistance replicon variants against ACH-806 was performed to map the mutations conferring resistance to ACH-806 and to determine cross resistance profiles with other classes of HCV inhibitors. Several clones emerged after the addition of ACH-806 to HCV replicon cells at frequencies and durations similar to that observed with NS3 protease inhibitors and NS5B polymerase inhibitors. Phenotypic analyses of these clones revealed that they are resistant to ACH-806 but remain sensitive to other classes of HCV inhibitors. Moreover, no significant change in the susceptibility to ACH-806 was found when the replicon cellular clones resistant to NS3 protease inhibitors and NS5B polymerase inhibitors were examined. Sequencing of the entire coding region of ACH-806 resistant replicon variants yielded several consensus mutations. Reverse genetics identified two single mutations in NS3, a cysteine-to-serine mutation at amino acid 16 and an alanine-to-valine mutation at amino acid 39, that are responsible for the resistance of the replicon variants to ACH-806. Both mutations are located at the N terminus of NS3 where extensive interactions with the central hydrophobic region of NS4A exist. These data provide evidence that ACH-806 inhibits HCV replication by a novel mechanism. PMID- 18411326 TI - Activity of pradofloxacin against Porphyromonas and Prevotella spp. Implicated in periodontal disease in dogs: susceptibility test data from a European multicenter study. AB - Collaborating veterinarians from five European countries collected subgingival bacterial samples from dogs exhibiting clinical periodontal disease. Sterile endodontic paper points were used for collection of the samples, which were transported to a central laboratory for susceptibility testing. Anaerobic bacteria were isolated and Porphyromonas and Prevotella isolates identified to the species level; susceptibility to pradofloxacin and metronidazole was determined using the CLSI agar dilution methodology. A total of 630 isolates, 310 of Porphyromonas spp. and 320 of Prevotella spp., were isolated. Pradofloxacin MIC data for all isolates were in the range of < or =0.016 to 1 microg/ml, the overall MIC(50) was 0.062, and the overall MIC(90) was 0.25 microg/ml. There were no differences in activity against Porphyromonas and Prevotella isolates or in the pradofloxacin susceptibility distributions from the different European countries. All isolates were within the wild-type distribution and were fully susceptible to pradofloxacin. Metronidazole was also highly active against these strains: 316 of 320 Prevotella strains (98.8%) and 309 of 310 Porphyromonas strains (99.7%) were susceptible (MICs of < or =8 microg/ml). However, three Prevotella strains had intermediate metronidazole susceptibility (MICs of 16 microg/ml), while one Prevotella and one Porphyromonas strain were metronidazole resistant (MICs of 128 and 256 microg/ml, respectively). Pradofloxacin, a novel broad-spectrum fluoroquinolone, demonstrates a high degree of antianaerobic activity against strains isolated from clinical cases of periodontal disease and shows activity against metronidazole-resistant isolates. The broad-spectrum activity of pradofloxacin makes it a suitable candidate for the treatment of periodontal disease in dogs. PMID- 18411325 TI - Discordant patterns of genetic variation at two chloroquine resistance loci in worldwide populations of the malaria parasite Plasmodium falciparum. AB - Mutations in the chloroquine resistance (CQR) transporter gene of Plasmodium falciparum (Pfcrt; chromosome 7) play a key role in CQR, while mutations in the multidrug resistance gene (Pfmdr1; chromosome 5) play a significant role in the parasite's resistance to a variety of antimalarials and also modulate CQR. To compare patterns of genetic variation at Pfcrt and Pfmdr1 loci, we investigated 460 blood samples from P. falciparum-infected patients from four Asian, three African, and three South American countries, analyzing microsatellite (MS) loci flanking Pfcrt (five loci [approximately 40 kb]) and Pfmdr1 (either two loci [approximately 5 kb] or four loci [approximately 10 kb]). CQR Pfmdr1 allele associated MS haplotypes showed considerably higher genetic diversity and higher levels of subdivision than CQR Pfcrt allele-associated MS haplotypes in both Asian and African parasite populations. However, both Pfcrt and Pfmdr1 MS haplotypes showed similar levels of low diversity in South American parasite populations. Median-joining network analyses showed that the Pfcrt MS haplotypes correlated well with geography and CQR Pfcrt alleles, whereas there was no distinct Pfmdr1 MS haplotype that correlated with geography and/or CQR Pfmdr1 alleles. Furthermore, multiple independent origins of CQR Pfmdr1 alleles in Asia and Africa were inferred. These results suggest that variation at Pfcrt and Pfmdr1 loci in both Asian and African parasite populations is generated and/or maintained via substantially different mechanisms. Since Pfmdr1 mutations may be associated with resistance to artemisinin combination therapies that are replacing CQ, particularly in Africa, it is important to determine if, and how, the genetic characteristics of this locus change over time. PMID- 18411327 TI - A role for DPPX modulating external TEA sensitivity of Kv4 channels. AB - Shal-type (Kv4) channels are expressed in a large variety of tissues, where they contribute to transient voltage-dependent K+ currents. Kv4 are the molecular correlate of the A-type current of neurons (I(SA)), the fast component of I(TO) current in the heart, and also of the oxygen-sensitive K+ current (K(O2)) in rabbit carotid body (CB) chemoreceptor cells. The enormous degree of variability in the physiological properties of Kv4-mediated currents can be attributable to the complexity of their regulation together with the large number of ancillary subunits and scaffolding proteins that associate with Kv4 proteins to modify their trafficking and their kinetic properties. Among those, KChIPs and DPPX proteins have been demonstrated to be integral components of I(SA) and I(TO) currents, as their coexpression with Kv4 subunits recapitulates the kinetics of native currents. Here, we explore the presence and functional contribution of DPPX to K(O2) currents in rabbit CB chemoreceptor cells by using DPPX functional knockdown with siRNA. Additionally, we investigate if the presence of DPPX endows Kv4 channels with new pharmacological properties, as we have observed anomalous tetraethylammonium (TEA) sensitivity in the native K(O2) currents. DPPX association with Kv4 channels induced an increased TEA sensitivity both in heterologous expression systems and in CB chemoreceptor cells. Moreover, TEA application to Kv4-DPPX heteromultimers leads to marked kinetic effects that could be explained by an augmented closed-state inactivation. Our data suggest that DPPX proteins are integral components of K(O2) currents, and that their association with Kv4 subunits modulate the pharmacological profile of the heteromultimers. PMID- 18411328 TI - Antibody-based validation of CNS ion channel drug targets. PMID- 18411329 TI - How highly charged anionic lipids bind and regulate ion channels. PMID- 18411330 TI - How to validate a heteromeric ion channel drug target: assessing proper expression of concatenated subunits. PMID- 18411331 TI - Ion channels as drug targets: the next GPCRs. PMID- 18411333 TI - Clearance of extracellular K+ during muscle contraction--roles of membrane transport and diffusion. AB - Excitation of muscle often leads to a net loss of cellular K + and a rise in extracellular K+([K+]o), which in turn inhibits excitability and contractility. It is important, therefore, to determine how this K+ is cleared by diffusion into the surroundings or by reaccumulation into the muscle cells. The inhibitory effects of the rise in [K+] o may be assessed from the time course of changes in tetanic force in isolated muscles where diffusional clearance of K+ is eliminated by removing the incubation medium and allowing the muscles to contract in air. Measurements of tetanic force, endurance, and force recovery showed that in rat soleus and extensor digitorum longus (EDL) muscles there was no significant difference between the performance of muscles contracting in buffer or in air for up to 8 min. Ouabain-induced inhibition of K+ clearance via the Na+,K+ pumps markedly reduced contractile endurance and force recovery in air. Incubation in buffer containing 10 mM K+ clearly inhibited force development and endurance,and these effects were considerably reduced by stimulating Na+,K+ pumps with the 2 agonist salbutamol. Following 30-60 s of continuous stimulation at 60 Hz, the amount of K + released into the extracellular space was assessed from washout experiments. The release of intracellular K+ per pulse was fourfold larger in EDL than in soleus,and in the two muscles, the average [K+] o reached 52.4 and 26.0 mM, respectively, appreciably higher than previously detected. In conclusion, prevention of diffusion of K+ from the extracellular space of isolated working muscles causes only modest interference with contractile performance. The Na+,K+ pumps play a major role in the clearance of K+ and the maintenance of force. This new information is important for the evaluation of K+ -induced inhibition in muscles, where diffusional clearance of K+ is reduced by tension development sufficient to suppress circulation. PMID- 18411334 TI - Perspectives on how to drug an ion channel. PMID- 18411332 TI - Lipid bilayer-mediated regulation of ion channel function by amphiphilic drugs. PMID- 18411335 TI - Implementing safe routes to school: application for the socioecological model and issues to consider. AB - The newly established national Safe Routes to School (SRTS) program has the potential to positively influence individuals, communities, and the environment regardless of race, ethnicity, or socioeconomic status. Many communities are applying their interest in physical activity promotion toward creating policies and programs to encourage active travel, though many barriers exist. SRTS legislation provides funds to address some of the barriers and improve the ability of students to safely walk and bicycle to school. SRTS requires that 70% to 90% of the funds be used for infrastructure projects (i.e., engineering treatments, such as sidewalk construction), and 10% to 30% for noninfrastructure activities, such as education, encouragement, and enforcement. The socioecological model (SEM) is widely used in public health and includes five levels of influence on behavior, from individual to public policy. Application of the SEM to SRTS provides a framework for a comprehensive approach to improve active travel to school. PMID- 18411336 TI - The 6-step model for community empowerment: revisited in public housing communities for low-income senior citizens. AB - The 6-step community empowerment model was replicated in communities with different geographical, racial, and age backgrounds from the original application. Resident groups of Blue Ribbon Health Panels (BRHPs) in federally funded senior housing in Pennsylvania followed the 6 steps to identify community health issues, to develop strategies to address priority issues, and to implement the strategies in collaboration with partner agencies. The 6-step model served as an operationalization strategy of community empowerment by facilitating quick accomplishments of communities' desired outcomes, legitimizing and motivating BRHP efforts. Community capacities to actively participate and collaborate influenced the model's progress in this replication study, as did partner agencies' capacities to adhere to the community-based participatory and collaborative orientation of the project. Community capacity development and partnership facilitation would be important for a community empowerment project, as well as consistent and clear communication among everyone involved in the process. PMID- 18411337 TI - TL1A-DR3 interaction regulates Th17 cell function and Th17-mediated autoimmune disease. AB - T helper type 17 (Th17) cells play an important pathogenic function in autoimmune diseases; their regulation, however, is not well understood. We show that the expression of a tumor necrosis factor receptor family member, death receptor 3 (DR3; also known as TNFRSF25), is selectively elevated in Th17 cells, and that TL1A, its cognate ligand, can promote the proliferation of effector Th17 cells. To further investigate the role of the TL1A-DR3 pathway in Th17 regulation, we generated a TL1A-deficient mouse and found that TL1A(-/-) dendritic cells exhibited a reduced capacity in supporting Th17 differentiation and proliferation. Consistent with these data, TL1A(-/-) animals displayed decreased clinical severity in experimental autoimmune encephalomyelitis (EAE). Finally, we demonstrated that during EAE disease progression, TL1A was required for the optimal differentiation as well as effector function of Th17 cells. These observations thus establish an important role of the TL1A-DR3 pathway in promoting Th17 cell function and Th17-mediated autoimmune disease. PMID- 18411338 TI - Regulation of inflammatory responses by IL-17F. AB - Although interleukin (IL) 17 has been extensively characterized, the function of IL-17F, which has an expression pattern regulated similarly to IL-17, is poorly understood. We show that like IL-17, IL-17F regulates proinflammatory gene expression in vitro, and this requires IL-17 receptor A, tumor necrosis factor receptor-associated factor 6, and Act1. In vivo, overexpression of IL-17F in lung epithelium led to infiltration of lymphocytes and macrophages and mucus hyperplasia, similar to observations made in IL-17 transgenic mice. To further understand the function of IL-17F, we generated and analyzed mice deficient in IL 17F or IL-17. IL-17, but not IL-17F, was required for the initiation of experimental autoimmune encephalomyelitis. Mice deficient in IL-17F, but not IL 17, had defective airway neutrophilia in response to allergen challenge. Moreover, in an asthma model, although IL-17 deficiency reduced T helper type 2 responses, IL-17F-deficient mice displayed enhanced type 2 cytokine production and eosinophil function. In addition, IL-17F deficiency resulted in reduced colitis caused by dextran sulfate sodium, whereas IL-17 knockout mice developed more severe disease. Our results thus demonstrate that IL-17F is an important regulator of inflammatory responses that seems to function differently than IL-17 in immune responses and diseases. PMID- 18411339 TI - Bmi1 regulates memory CD4 T cell survival via repression of the Noxa gene. AB - The maintenance of memory T cells is central to the establishment of immunological memory, although molecular details of the process are poorly understood. In the absence of the polycomb group (PcG) gene Bmi1, the number of memory CD4(+) T helper (Th)1/Th2 cells was reduced significantly. Enhanced cell death of Bmi1(-/-) memory Th2 cells was observed both in vivo and in vitro. Among various proapoptotic genes that are regulated by Bmi1, the expression of proapoptotic BH3-only protein Noxa was increased in Bmi1(-/-) effector Th1/Th2 cells. The generation of memory Th2 cells was restored by the deletion of Noxa, but not by Ink4a and Arf. Direct binding of Bmi1 to the Noxa gene locus was accompanied by histone H3-K27 methylation. The recruitment of other PcG gene products and Dnmt1 to the Noxa gene was highly dependent on the expression of Bmi1. In addition, Bmi1 was required for DNA CpG methylation of the Noxa gene. Moreover, memory Th2-dependent airway inflammation was attenuated substantially in the absence of Bmi1. Thus, Bmi1 controls memory CD4(+) Th1/Th2 cell survival and function through the direct repression of the Noxa gene. PMID- 18411340 TI - Native and aspirin-triggered lipoxins control innate immunity by inducing proteasomal degradation of TRAF6. AB - Innate immune signaling is critical for the development of protective immunity. Such signaling is, perforce, tightly controlled. Lipoxins (LXs) are eicosanoid mediators that play key counterregulatory roles during infection. The molecular mechanisms underlying LX-mediated control of innate immune signaling are of interest. In this study, we show that LX and aspirin (ASA)-triggered LX (ATL) inhibit innate immune signaling by inducing suppressor of cytokine signaling (SOCS) 2-dependent ubiquitinylation and proteasome-mediated degradation of TNF receptor-associated factor (TRAF) 2 and TRAF6, which are adaptor molecules that couple TNF and interleukin-1 receptor/Toll-like receptor family members to intracellular signaling events. LX-mediated degradation of TRAF6 inhibits proinflammatory cytokine production by dendritic cells. This restraint of innate immune signaling can be ablated by inhibition of proteasome function. In vivo, this leads to dysregulated immune responses, accompanied by increased mortality during infection. Proteasomal degradation of TRAF6 is a central mechanism underlying LX-driven immune counterregulation, and a hitherto unappreciated mechanism of action of ASA. These findings suggest a new molecular target for drug development for diseases marked by dysregulated inflammatory responses. PMID- 18411341 TI - Essential role of TNF receptor superfamily 25 (TNFRSF25) in the development of allergic lung inflammation. AB - We identify the tumor necrosis factor receptor superfamily 25 (TNFRSF25)/TNFSF15 pair as critical trigger for allergic lung inflammation, which is a cardinal feature of asthma. TNFRSF25 (TNFR25) signals are required to exert T helper cell 2 (Th2) effector function in Th2-polarized CD4 cells and co-stimulate interleukin (IL)-13 production by glycosphingolipid-activated NKT cells. In vivo, antibody blockade of TNFSF15 (TL1A), which is the ligand for TNFR25, inhibits lung inflammation and production of Th2 cytokines such as IL-13, even when administered days after airway antigen exposure. Similarly, blockade of TNFR25 by a dominant-negative (DN) transgene, DN TNFR25, confers resistance to lung inflammation in mice. Allergic lung inflammation-resistant, NKT-deficient mice become susceptible upon adoptive transfer of wild-type NKT cells, but not after transfer of DN TNFR25 transgenic NKT cells. The TNFR25/TL1A pair appears to provide an early signal for Th2 cytokine production in the lung, and therefore may be a drug target in attempts to attenuate lung inflammation in asthmatics. PMID- 18411342 TI - Robert Menard: Tailing malaria parasites to the red blood cell. Interviewed by Hema Bashyam. PMID- 18411343 TI - Biological ageing and cardiovascular disease. PMID- 18411344 TI - B-type natriuretic peptide: a treatment, not a diagnostic marker. PMID- 18411345 TI - Identification of multiple plaque ruptures by optical coherence tomography in a patient with acute myocardial infarction: a three-vessel study. PMID- 18411346 TI - Natriuretic peptides or psychometric tests? Prognostic markers in congestive heart failure. PMID- 18411347 TI - Closure devices for femoral punctures. PMID- 18411349 TI - A heartless patient. PMID- 18411350 TI - The heart rate hypothesis: ready to be tested. AB - There is increasing evidence that increased heart rate may be an independent risk factor for cardiovascular morbidity and mortality both in patients with ischaemic heart disease and in the general population. Elevated heart rate in coronary artery disease is a major determinant of oxygen consumption and appears to evoke most episodes of ischaemia. Increased resting heart rate may also contribute to the development of atherosclerosis, facilitate plaque destabilisation and initiate arrhythmias, leading to acute coronary events and sudden death. Reducing heart rate is a central aim in the treatment of stable angina pectoris; this therapeutic approach may have an essential role in lowering the incidence of cardiovascular morbidity and mortality in patients with pre-existing ischaemic heart disease. However, this heart rate hypothesis has not thus far been proven. Evidence suggests that the use of heart rate-lowering drugs may have a beneficial effect; however, most treatments for angina have additional negative inotropic effects on the heart. This hypothesis can now be tested following the recent development of selective heart rate drugs. PMID- 18411351 TI - Failed endothelialisation of a percutaneous atrial septal defect closure device. PMID- 18411352 TI - Two- and three-dimensional imaging of coarctation shelf in the human fetus. PMID- 18411353 TI - Coil embolisation for diffuse pulmonary arteriovenous malformations. PMID- 18411354 TI - Ezetimibe for the treatment of primary (heterozygous-familial and non-familial) hypercholesterolaemia: NICE technology appraisal guidance. PMID- 18411355 TI - Commentary on NICE technology appraisal guidance on ezetimibe for the treatment of primary (heterozygous-familial and non-familial) hypercholesterolaemia. PMID- 18411357 TI - Antibiotic prophylaxis in cardiac surgery. PMID- 18411359 TI - Prognostic value of LV long-axis versus short-axis systolic function in heart failure. PMID- 18411360 TI - Troubleshooting implantable cardioverter-defibrillator related problems. PMID- 18411361 TI - Bleeding and transfusion in acute coronary syndromes: a shift in the paradigm. PMID- 18411362 TI - Radiation exposure in invasive cardiology. PMID- 18411364 TI - Nicorandil-induced colonic ulceration. PMID- 18411365 TI - Fat chance. PMID- 18411366 TI - Why does this controversy still exist? PMID- 18411367 TI - Perception versus reality: overcoming barriers to colorectal cancer screening. PMID- 18411368 TI - A weighty issue: medication as a cornerstone of medical obesity management. PMID- 18411370 TI - Should Canadians be offered systematic screening for colorectal cancer?: yes. PMID- 18411371 TI - Should Canadians be offered systematic screening for colorectal cancer?: no. PMID- 18411374 TI - [Do family physicians make enough house calls?]. PMID- 18411375 TI - Preventive guidelines. PMID- 18411376 TI - Patient-friendly guidelines. PMID- 18411377 TI - Lead exposure during breastfeeding. AB - QUESTION: Owing to the recent concerns of lead (Pb) leaking into tap water, one of our female patients is concerned about the effects of Pb exposure to newborns while breastfeeding. How should I advise her and should she switch to formula feeding? ANSWER: Lead exposure through drinking tap water while breastfeeding is not associated with any serious concerns in most available studies. There is currently no safe level of Pb exposure, but environmental exposure within Canada is low. At present, Pb levels in drinking water are carefully monitored by Health Canada and are not likely to be of concern to breastfeeding mothers. Switching to formula feeding is not necessary and not recommended, as improperly prepared formula can have higher Pb levels. PMID- 18411378 TI - Dermacase. Atypical fibroxanthoma. PMID- 18411379 TI - Practice tips. Ethnic elderly with dementia: overcoming the cultural barriers to their care. PMID- 18411380 TI - Management of bone metastases. PMID- 18411383 TI - Teaching pharmacotherapeutics to family medicine residents: a curriculum. AB - PROBLEM BEING ADDRESSED: Medication prescribing is becoming increasingly complex, and the need for formal curricula in pharmacotherapeutics and medication prescribing in accredited family medicine residency programs has been advocated. OBJECTIVE OF PROGRAM: The main objective of the pharmacotherapeutic curriculum is to support the development of family medicine residents' pharmacotherapeutic knowledge and medication prescribing skills required for rational prescribing. PROGRAM DESCRIPTION: The curriculum has 4 main components: 1) a medication prescribing framework based on the main tasks and key decisions related to the prescribing of medications, 2) 12 pharmacotherapeutic topics identified in the needs assessment, 3) a 5-step process for session design used by the curriculum development team, and 4) a description of specific roles of facilitators involved in delivering the curriculum. Formative evaluation of the curriculum using resident focus groups has helped to inform the further development of its components. CONCLUSION: A formalized curriculum was created to build knowledge of pharmacotherapeutics and effective medication prescribing skills, which are necessary for the current complex environment of patient care and medication management. PMID- 18411381 TI - Vitamin B12 and health. AB - OBJECTIVE: To review recent evidence that suggests vitamin B12 is associated with risk reduction for some chronic diseases and birth defects. QUALITY OF EVIDENCE: A MEDLINE search from 1999 to 2007 was performed using the key word vitamin B12. The most relevant articles (129) dealt with cardiovascular disease, cancer, mental health, and birth outcomes;most studies presented level II evidence. MAIN MESSAGE: Vitamin B12 might confer health benefits; however, such benefits are difficult to ascertain because of the complementary functions of vitamin B12 and folic acid. Vitamin B12 might lower high homocysteine levels below a threshold level achieved by folic acid alone. Furthermore, the interactions between the nutritional environment and genotype might have an important influence on vitamin B12, chronic disease risk, and risk of neural tube defects. CONCLUSION: Vitamin B12 might help protect against chronic disease and neural tube defects, but more research, particularly in the area of nutritional genomics, is needed to determine how vitamin B12 might augment the benefits of folic acid. Some consideration should be given to the potential value of fortifying foods with vitamin B12 in addition to the current mandatory folic acid fortification of grains. PMID- 18411382 TI - Role for a sense of self-worth in weight-loss treatments: helping patients develop self-efficacy. AB - OBJECTIVE: To recommend strategies for enhancing patients' sense of self-worth and self-efficacy in order to give them sufficient faith in themselves to make healthier choices about their weight. SOURCES OF INFORMATION: PubMed, PsycINFO, Google Scholar, and APA Journals Online were searched for original research articles on treatment models and outcome review articles from 1960 to the present. Key search terms were weight loss, weight-loss treatments, diets and weight loss, psychology and obesity, physiology and obesity, and exercise and weight loss. Most evidence was level I and level II. MAIN MESSAGE: In spite of extensive research, there is widespread belief that the medical system has failed to stem the tide of weight gain in North America. The focus has been on physiologic, behavioural, and cultural explanations for what is seen as a relatively recent phenomenon, while the self-perception of overweight individuals has been largely overlooked. Professional treatments have consisted mainly of cognitive behavioural therapies and rest on the premise that overweight patients will effectively apply the cognitive behavioural therapy principles. In the long run, professional and commercial programs are often ineffective. We need treatments that include strategies to repair ego damage, enhance the sense of self-worth, and develop self-efficacy so that overweight patients can become the agents of change in their pursuit of well-being. CONCLUSION: Self-efficacy correlates positively with success in all realms of personal endeavour, and we can help our overweight patients become more self-reliant. PMID- 18411384 TI - Can family physicians help patients initiate basal insulin therapy successfully?: randomized trial of patient-titrated insulin glargine compared with standard oral therapy: lessons for family practice from the Canadian INSIGHT trial. AB - OBJECTIVE: To determine whether FPs could help patients implement bedtime basal insulin therapy as successfully as diabetes experts could. DESIGN: National, multicentre, randomized, open-label trial designed to assess use of bedtime basal insulin therapy compared with use of standard oral-agent therapy for patients with type 2 diabetes being treated by diabetes experts or FPs. SETTING: Nineteen endocrinologist or expert sites and 34 family practices. PARTICIPANTS: A total of 405 adult patients with hemoglobin A1c (HbA1c) values of 7.5% to 11.0% who were taking 0 to 2 oral agents. INTERVENTION: Participants were randomized to receive either basal insulin therapy using glargine self-titrated according to a patient algorithm or conventional therapy with physician-adjusted doses of oral agents for a period of 24 weeks. MAIN OUTCOME MEASURES: The primary outcome was time to achieve 2 consecutive HbA1c values < or = 6.5%. Secondary outcomes were the proportion of subjects who achieved these HbA1c values, a fasting plasma glucose level < or = 5.5 mmol/L, and 2 consecutive HbA1c values < or = 7.0%; incidence, rate, and severity of hypoglycemia; daily variations in blood-glucose levels; and participants' lipid profiles. Post-hoc analysis sought to determine whether patients' outcomes differed in terms of the above measures depending on whether they had been treated by diabetes experts or FPs. RESULTS: A total of 206 patients were randomized to the glargine group, and 199 to the oral agents group. In total, 145 patients were followed by experts and 260 by FPs. Mean reductions in HbA1c and fasting plasma glucose levels and rates of hypoglycemia were comparable in the 2 groups. Patients of both types of physicians achieved significantly greater reductions in fasting plasma glucose with glargine than with oral agents (FPs: -4.14 vs -2.45 mmol/L, P = .0001; experts: -3.47 vs -2.19 mmol/L, P = .0013). Patients of FPs achieved significantly greater reductions in HbA1c levels with glargine than with oral agents (FPs: -1.64 vs -1.26%, P = .0058; experts: -1.41 vs-1.24%, P = .3331). Final mean insulin doses were higheramong FPs' patients than among experts' patients (41.74vs 31.66 units, P = .015). Family physicians were more aggressive in their use of insulin, while experts used more oral agents. There were no significant differences inefficacy of treatment. CONCLUSION: In most settings, FPs could easily implement the patient-driven bedtime basal insulin protocol used in this study. PMID- 18411385 TI - The evidence is in. Why are IUDs still out?: family physicians' perceptions of risk and indications. AB - OBJECTIVE: Intrauterine devices (IUDs) for contraception are used infrequently in Canada despite their well-studied safety and efficacy. The purpose of this study was to investigate FPs' perceptions of the risks of and indications for using IUDs, as these perceptions might be a key factor in why IUDs are underused. DESIGN: Mailed survey. SETTING: Kingston, a midsized city in Ontario. PARTICIPANTS: All FPs practising in Kingston. MAIN OUTCOME MEASURES: The primary outcome was the proportion of FPs who ranked the importance of risks of and contraindications to using IUDs in keeping with rankings in established clinical guidelines.Secondary outcomes were FP IUD prescription and insertion patterns, their perceptions of patients' access to physicians who insert IUDs, and their interest in a course on IUD insertion. RESULTS: Response rate was 81%. Contrary to the evidence, more than 60% of FPs thought pelvic inflammatory disease and ectopic pregnancy were major risks, and nearly half thought failure of IUDs was a major risk. Fewer than one-third would recommend IUDs as an option for nulliparous women, for postcoital contraception, for women with moderate-sized fibroids, or for women with pelvic inflammatory disease during the last year,though none of these are contraindications according to established clinical guidelines. About 82% of FPs prescribed IUDs, and 41% inserted them. Newer graduates were more likely than older graduates to prescribe only levonorgestrel IUDs (36% vs 8%). About 93% of survey respondents reported that patients' access to physicians who insert IUDs was not a factor in recommending IUDs to them. CONCLUSION: A substantial proportion of surveyed FPs believed that the side effects of IUDs were more severe than is supported by clinical guidelines and were misinformed about the range of women who could benefit from IUDs. These misconceptions likely contribute to the low rate of IUD use in Canada. PMID- 18411386 TI - Medical students' views on training in intellectual disabilities. AB - OBJECTIVE: To examine undergraduate medical training in the field of intellectual disabilities (ID) from the perspective of clinical clerks. DESIGN: Cross sectional self-administered survey. SETTING: Clerkship rotations at Queen's University in Kingston, Ont, and the University of Toronto in Ontario in 2006. PARTICIPANTS: A total of 196 upper-year undergraduate medical students (clerks). MAIN OUTCOME MEASURES: Contact with people with ID, training in the field of ID, perceptions of current training in ID, and views on ways to improve the curriculum in the area of ID. RESULTS: Most students (85.6%) had received some didactic and clinical training in managing patients with ID,but most of these (93.3%) believed that curriculum enhancements, especially more contact with patients with ID and more time in the curriculum for training in care of people with ID, were necessary. CONCLUSION: This study found that the curriculum enhancements long recommended by experts in the field of ID were also desired by clinical clerks. This finding adds considerable weight to the recommendation that improvements in training in ID should be incorporated into undergraduate medical education programs. PMID- 18411388 TI - Croup. PMID- 18411387 TI - Patients' perceptions and experiences of family medicine residents in the office. AB - OBJECTIVE: To examine patients' perceptions and experiences of having family medicine residents in the office. DESIGN: Descriptive survey; questionnaire completed by patients. SETTING: Outpatient office of 4 family physicians in the greater Vancouver area, affiliated with the Department of Family Medicine at the University of British Columbia. PARTICIPANTS: A total of 265 English-speaking adult patients attending the office. MAIN OUTCOME MEASURES: Patients' self reported perceptions and experiences of having family medicine residents in the office. RESULTS: Response rate was 94.7% (251 of 265 patients completed the questionnaire). Although 81% of respondents had seen residents in the office, 59% did not understand a resident's training or thought residents were medical students. The 3 main reasons participants gave for choosing to have residents involved in their care were the following: to contribute to training future doctors (61.8%); to obtain 2 opinions instead of 1 (20%); and residents are most up-to-date (11.2%). The most common reasons for choosing not to see residents were the following: to continue relationships with their own doctors (54.2%); to avoid the need to repeat history (18.6%); and the perception that residents are less experienced (16.9%). Having a resident involved in their care was perceived as a positive experience by 95.5% of respondents who had seen residents. Overall satisfaction with care and overall comfort in dealing with residents were ranked good to excellent by 91.8% and 90.8% of respondents, respectively. About 71% of patients said they would choose to have residents involved in their care. CONCLUSION: Respondents reported very positive experiences with having family medicine residents in the office. Overall comfort and satisfaction with seeing family medicine residents was reported to be extremely high, and most patients surveyed would choose to have family medicine residents involved in their care. Patients needed to know more about the resident's level of training and the role of residents in patient-resident interactions. PMID- 18411389 TI - Prevention of kernicterus: new guidelines and the critical role of family physicians. PMID- 18411390 TI - Journey toward change: a family practice's experience with program development. PMID- 18411391 TI - Are we over-doctoring?: thoughts on bedside manner. PMID- 18411392 TI - The kindness of strangers: an elective in Ghana. PMID- 18411395 TI - The X-factor. PMID- 18411399 TI - Drug transporter and cytochrome P450 mRNA expression in human ocular barriers: implications for ocular drug disposition. AB - Studies were designed to quantitatively assess the mRNA expression of 1) 10 cytochrome P450 (P450) enzymes in human cornea, iris-ciliary body (ICB), and retina/choroid relative to their levels in the liver, and of 2) 21 drug transporters in these tissues relative to their levels in human small intestine, liver, or kidney. Potential species differences in mRNA expression of PEPT1, PEPT2, and MDR1 were also assessed in these ocular tissues from rabbit, dog, monkey, and human. P450 expression was either absent or marginal in human cornea, ICB, and retina/choroid, suggesting a limited role for P450-mediated metabolism in ocular drug disposition. In contrast, among 21 key drug efflux and uptake transporters, many exhibited relative expression levels in ocular tissues comparable with those observed in small intestine, liver, or kidney. This robust ocular transporter presence strongly suggests a significant role that transporters may play in ocular barrier function and ocular pharmacokinetics. The highly expressed efflux transporter MRP1 and uptake transporters PEPT2, OCT1, OCTN1, and OCTN2 may be particularly important in absorption, distribution, and clearance of their drug substrates in the eye. Evidence of cross-species ocular transporter expression differences noted in these studies supports the conclusion that transporter expression variability, along with anatomic and physiological differences, should be taken into consideration to better understand animal ocular pharmacokinetic and pharmacodynamic data and the scalability to human for ocular drugs. PMID- 18411400 TI - Inhibitory effects of seven components of danshen extract on catalytic activity of cytochrome P450 enzyme in human liver microsomes. AB - The potential for herb-drug interactions has recently received greater attention worldwide, considering the fact that the use of herbal products becomes more and more widespread. The goal of this work was to examine the potential for the metabolism-based drug interaction arising from seven active components (danshensu, protocatechuic aldehyde, protocatechuic acid, salvianolic acid B, tanshinone I, tanshinone IIA, and cryptotanshinone) of danshen extract. Probe substrates of cytochrome P450 enzymes were incubated in human liver microsomes (HLMs) with or without each component of danshen extract. IC(50) and K(i) values were estimated, and the types of inhibition were determined. Among the seven components of danshen extract, tanshinone I, tanshinone IIA, and cryptotanshinone were potent competitive inhibitors of CYP1A2 (K(i) = 0.48, 1.0, and 0.45 microM, respectively); danshensu was a competitive inhibitor of CYP2C9 (K(i) = 35 microM), and cryptotanshinone was a moderate mixed-type inhibitor of CYP2C9 (K(i) = 8 microM); cryptotanshinone inhibited weakly and in mixed mode against CYP2D6 activity (K(i) = 68 microM), and tanshinone I was a weak inhibitor of CYP2D6 (IC(50) = 120 microM); and protocatechuic aldehyde was a weak inhibitor of CYP3A4 (IC(50) = 130 and 160 microM for midazolam and testosterone, respectively). These findings provided some useful information for safe and effective use of danshen preparations in clinical practice. Our data indicated that it was necessary to study the in vivo interactions between drugs and pharmaceuticals with danshen extract. PMID- 18411401 TI - Hepatocellular binding of drugs: correction for unbound fraction in hepatocyte incubations using microsomal binding or drug lipophilicity data. AB - Analogous to the fraction unbound in microsomes (fu(mic)), fraction unbound in hepatocyte incubations (fu(hep)) is an important parameter in the prediction of intrinsic clearance and potential drug-drug interactions. A recent study by Austin et al. (Drug Metab Dispos 33:419-425, 2005) proposed a linear 1:1 relationship between the extent of binding to microsomes at 1 mg/ml and to hepatocytes at 10(6) million cells/ml. The current study collates a fu(mic) and fu(hep) database for 39 drugs to examine the relationship between binding in microsomes and hepatocytes. A new nonlinear empirical equation is proposed as an alternative to the linear relationship to relate binding between the two systems. The nonlinear equation results in higher prediction accuracy and lower bias in comparison to the linear relationship, in particular for drugs with fu(hep) < 0.4. The proposed equation is further extended to allow direct prediction of fu(hep) from drug lipophilicity data by substituting the fu(mic) term by the Hallifax and Houston predictive equation (Drug Metab Dispos 34:724-726, 2006). The prediction accuracy of this approach is high for relatively hydrophilic drugs (logP/D < or = 2.5), whereas less accurate prediction of the fu(hep) is observed for lipophilic drugs (logP > 3), consistent with the limitations observed for microsomal binding predictive tools. In conclusion, the proposed nonlinear equations provide an accurate predictive tool to estimate fu(hep) for the in vitro-in vivo extrapolation of intrinsic clearance and inhibition parameters. PMID- 18411402 TI - Bioactivation of flutamide metabolites by human liver microsomes. AB - Flutamide, a widely used nonsteroidal antiandrogen drug for the treatment of prostate cancer, has been associated with rare incidences of hepatotoxicity in patients. It is believed that bioactivation of flutamide and subsequent covalent binding to cellular proteins is responsible for its toxicity. A novel N-S glutathione adduct has been identified in a previous bioactivation study of flutamide (Kang et al., 2007). Due to the extensive first pass metabolism, flutamide metabolites such as 2-hydroxyflutamide and 4-nitro-3 (trifluoromethyl)phenylamine (Flu-1) have achieved plasma concentrations higher than the parent in prostate cancer patients. In vitro studies in human liver microsomes were conducted to probe the cytochrome P450 (P450)-mediated bioactivation of flutamide metabolites and identify the possible reactive species using reduced glutathione (GSH) as a trapping agent. Several GSH adducts (G1, Flu 1-G1, Flu-1-G2, Flu-6-Gs) derived from the metabolites of flutamide were identified and characterized. A comprehensive bioactivation mechanism was proposed to account for the formation of the observed GSH adducts. Of interest were the formation of a reactive intermediate by the desaturation of the isopropyl group of M5 and the unusual bioactivation of Flu-1. Studies using recombinant P450s suggested that the major P450 isozymes involved in the bioactivation of flutamide and its metabolites were CYP1A2, CYP3A4, and CYP2C19. These findings suggested that, in addition to the direct bioactivation of flutamide, the metabolites of flutamide could also be bioactivated and contribute to flutamide-induced hepatotoxicity. PMID- 18411403 TI - The mibefradil derivative NNC55-0396, a specific T-type calcium channel antagonist, exhibits less CYP3A4 inhibition than mibefradil. AB - A novel mibefradil derivative, NNC55-0396, designed to be hydrolysis-resistant, was shown to be a selective T-type Ca(2+) channel inhibitor without L-type Ca(2+) channel efficacy. However, its effects on cytochromes P450 (P450s) have not previously been examined. We investigated the inhibitory effects of NNC55-0396 toward seven major recombinant human P450s--CYP3A4, CYP2D6, CYP1A2, CYP2C9, CYP2C8, CYPC19, and CYP2E1--and compared its effects with those of mibefradil and its hydrolyzed metabolite, Ro40-5966. Our results show that CYP3A4 and CYP2D6 are the two P450s most affected by mibefradil, Ro40-5966, and NNC55-0396. Mibefradil (IC(50) = 33 +/- 3 nM, K(i) = 23 +/- 0.5 nM) and Ro40-5966 (IC(50) = 30 +/- 7.8 nM, K(i) = 21 +/- 2.8 nM) have a 9- to 10-fold greater inhibitory activity toward recombinant CYP3A4 benzyloxy-4-trifluoromethylcoumarin-O-debenzylation activity than NNC55-0396 (IC(50) = 300 +/- 30 nM, K(i) = 210 +/- 6 nM). More dramatically, mibefradil (IC(50) = 566 +/- 71 nM, K(i) = 202 +/- 39 nM) shows 19-fold higher inhibition of CYP3A-associated testosterone 6beta-hydroxylase activity in human liver microsomes compared with NNC55-0396 (IC(50) = 11 +/- 1.1 microM, K(i) = 3.9 +/- 0.4 microM). Loss of testosterone 6beta-hydroxylase activity by recombinant CYP3A4 was shown to be time- and concentration-dependent with both compounds. However, NNC55-0396 (K(I) = 3.87 microM, K(inact) = 0.061/min) is a much less potent mechanism-based inhibitor than mibefradil (K(I) = 83 nM, K(inact) = 0.048/min). In contrast, NNC55-0396 (IC(50) = 29 +/- 1.2 nM, K(i) = 2.8 +/- 0.3 nM) and Ro40-5966 (IC(50) = 46 +/- 11 nM, K(i) = 4.5 +/- 0.02 nM) have a 3- to 4 fold greater inhibitory activity toward recombinant CYP2D6 than mibefradil (IC(50) = 129 +/- 21 nM, K(i) = 12.7 +/- 0.9 nM). Our results suggest that NNC55 0396 could be a more favorable T-type Ca(2+) antagonist than its parent compound, mibefradil, which was withdrawn from the market because of strong inhibition of CYP3A4. PMID- 18411404 TI - A high-resolution map of nucleosome positioning on a fission yeast centromere. AB - A key element for defining the centromere identity is the incorporation of a specific histone H3, CENPA, known as Cnp1p in Schizosaccharomyces pombe. Previous studies have suggested that functional S. pombe centromeres lack regularly positioned nucleosomes and may involve chromatin remodeling as a key step of kinetochore assembly. We used tiling microarrays to show that nucleosomes are, in fact, positioned in regular intervals in the core of centromere 2, providing the first high-resolution map of regional centromere chromatin. Nucleosome locations are not disrupted by mutations in kinetochore protein genes cnp1, mis18, mis12, nuf2, mal2; overexpression of cnp1; or the deletion of ams2, which encodes a GATA like factor participating in CENPA incorporation. Bioinformatics analysis of the centromere sequence indicates certain enriched motifs in linker regions between nucleosomes and reveals a sequence bias in nucleosome positioning. In addition, sequence analysis of nucleosome-free regions identifies novel binding sites of Ams2p. We conclude that centromeric nucleosome positions are stable and may be derived from the underlying DNA sequence. PMID- 18411405 TI - Population genetic analysis of shotgun assemblies of genomic sequences from multiple individuals. AB - We introduce a simple, broadly applicable method for obtaining estimates of nucleotide diversity from genomic shotgun sequencing data. The method takes into account the special nature of these data: random sampling of genomic segments from one or more individuals and a relatively high error rate for individual reads. Applying this method to data from the Celera human genome sequencing and SNP discovery project, we obtain estimates of nucleotide diversity in windows spanning the human genome and show that the diversity to divergence ratio is reduced in regions of low recombination. Furthermore, we show that the elevated diversity in telomeric regions is mainly due to elevated mutation rates and not due to decreased levels of background selection. However, we find indications that telomeres as well as centromeres experience greater impact from natural selection than intrachromosomal regions. Finally, we identify a number of genomic regions with increased or reduced diversity compared with the local level of human-chimpanzee divergence and the local recombination rate. PMID- 18411406 TI - Transcription factor and microRNA motif discovery: the Amadeus platform and a compendium of metazoan target sets. AB - We present a threefold contribution to the computational task of motif discovery, a key component in the effort of delineating the regulatory map of a genome: (1) We constructed a comprehensive large-scale, publicly-available compendium of transcription factor and microRNA target gene sets derived from diverse high throughput experiments in several metazoans. We used the compendium as a benchmark for motif discovery tools. (2) We developed Amadeus, a highly efficient, user-friendly software platform for genome-scale detection of novel motifs, applicable to a wide range of motif discovery tasks. Amadeus improves upon extant tools in terms of accuracy, running time, output information, and ease of use and is the only program that attained a high success rate on the metazoan compendium. (3) We demonstrate that by searching for motifs based on their genome-wide localization or chromosomal distributions (without using a predefined target set), Amadeus uncovers diverse known phenomena, as well as novel regulatory motifs. PMID- 18411407 TI - What can we learn from gene expression profiling of mouse oocytes? AB - Mammalian ooplasm supports the preimplantation development and reprograms the introduced nucleus transferred from a somatic cell to confer pluripotency in a cloning experiment. However, the underlying molecular mechanisms of oocyte competence remain unknown. Recent advances in microarray technologies have allowed gene expression profiling of such tiny specimens as oocytes and preimplantation embryos, generating a flood of information about gene expressions. So, what can we learn from it? Here, we review the initiative global gene expression studies of mouse and/or human oocytes, focusing on the lists of maternal transcripts and their expression patterns during oogenesis and preimplantation development. Especially, the genes expressed exclusively in oocytes should contribute to the uniqueness of oocyte competence, driving mammalian development systems of oocytes and preimplantation embryos. Furthermore, we discuss future directions for oocyte gene expression profiling, including discovering biomarkers of oocyte quality and exploiting the microarray data for 'making oocytes'. PMID- 18411408 TI - Expression of genes involved in prostaglandin E2 and progesterone production in bovine cumulus-oocyte complexes during in vitro maturation and fertilization. AB - Prostaglandin E(2) (PGE(2)) and progesterone appear to be critical mediators of cumulus expansion and the resumption of oocyte meiosis. The aim of this study was to identify the types of prostaglandin E synthase (PTGES) expressed in the bovine cumulus-oocyte complex (COC), to characterize their temporal expression during the periconceptional interval using an in vitro model of maturation (IVM) and fertilization (IVF), and to compare their expression with the level of steroidogenic gene expression. Real-time RT-PCR analysis revealed that enzymes related to the PGE(2) biosynthesis pathway were mainly expressed during IVM. Transcripts encoding PTGES1-3 were detected in bovine COCs. Only the expression of PTGES1 significantly increased during IVM whereas that of PTGES2 and PTGES3 remained unchanged. The induction of PTGES1 expression paralleled the induction of prostaglandin G/H synthase-2 (PTGS2) expression and the amounts of PGE(2) secreted by maturing COCs. Concomitantly, cholesterol side chain cleavage cytochrome P450 expression was significantly upregulated in maturing COCs and the high level of expression persisted in fertilized COCs. The expression of the StAR protein remained constant during IVM and then decreased significantly during IVF. Expression of the progesterone catabolic-related enzyme, 20alpha-hydroxysteroid dehydrogenase significantly decreased throughout the periconceptional interval. This was associated with a rising level of progesterone released by COCs in the culture media. In conclusion, our results suggest that the periconceptional differentiation of the bovine COC includes the transient induction of PGE(2) biosynthetic activity via the PTGS2/PTGES1 pathway during the maturation period and the increasing ability to produce progesterone from the immature to the fertilized stages. PMID- 18411409 TI - The interactions between cysteamine, cystine and cumulus cells increase the intracellular glutathione level and developmental capacity of goat cumulus denuded oocytes. AB - To improve in vitro maturation (IVM) of denuded oocytes (DOs), we observed the interactive effects of cysteamine, cystine and cumulus cells on the glutathione (L-gamma-glutamyl-L-cysteinyl-glycine; GSH) level and developmental capacity of goat IVM oocytes. Cysteamine supplementation increased the GSH level and blastocyst rates of both cumulus-oocyte complexes (COCs) and DOs, while the addition of cystine increased the GSH level and blastulation only in the presence of cumulus cells (COCs or DOs co-cultured on a cumulus cell monolayer). Simultaneous supplementation of cysteamine and cystine increased the GSH content and blastulation of co-cultured DOs to a level similar to that of COCs matured without thiol supplementation. Co-culture without thiol supplementation improved DOs' GSH synthesis but not blastulation. The results suggest that DOs cannot utilize cystine for GSH synthesis unless exogenous cysteamine is supplied by either cumulus cells or supplementation. Thus, while the addition of cystine alone is enough to improve IVM of COCs, improvement of DOs requires supplementation of both cystine and cysteamine. Synergic actions between cysteamine, cystine and cumulus cells restore the GSH level and developmental capacity of goat DOs. PMID- 18411410 TI - Expression of genes involved in early cell fate decisions in human embryos and their regulation by growth factors. AB - Little is understood about the regulation of gene expression in human preimplantation embryos. We set out to examine the expression in human preimplantation embryos of a number of genes known to be critical for early development of the murine embryo. The expression profile of these genes was analysed throughout preimplantation development and in response to growth factor (GF) stimulation. Developmental expression of a number of genes was similar to that seen in murine embryos (OCT3B/4, CDX2, NANOG). However, GATA6 is expressed throughout preimplantation development in the human. Embryos were cultured in IGF I, leukaemia inhibitory factor (LIF) or heparin-binding EGF-like growth factor (HBEGF), all of which are known to stimulate the development of human embryos. Our data show that culture in HBEGF and LIF appears to facilitate human embryo expression of a number of genes: ERBB4 (LIF) and LIFR and DSC2 (HBEGF) while in the presence of HBEGF no blastocysts expressed EOMES and when cultured with LIF only two out of nine blastocysts expressed TBN. These data improve our knowledge of the similarities between human and murine embryos and the influence of GFs on human embryo gene expression. Results from this study will improve the understanding of cell fate decisions in early human embryos, which has important implications for both IVF treatment and the derivation of human embryonic stem cells. PMID- 18411411 TI - Control of nuclear remodelling and subsequent in vitro development and methylation status of porcine nuclear transfer embryos. AB - We attempted to control the nuclear remodelling of somatic cell nuclear transfer embryos (NTs) and examined their subsequent development and DNA methylation patterns in pigs. Porcine foetal fibroblasts were fused to enucleated oocytes treated with either 5 mM caffeine for 2.5 h or 0.5 mM vanadate for 0.5 h. After activation, NTs were cultured in vitro for 6 days to examine their development. The nuclear remodelling type of the reconstituted embryos was evaluated 1 h after fusion. Methylated DNA of in vitro-fertilised (IVF) embryos and NTs at various developmental stages and of donor cells was detected using a 5-methylcytosine (5 MeC) antibody. Caffeine-treated NTs induced premature chromosome condensation at a high rate (P<0.05), whereas most vanadate-treated NTs formed a pronucleus-like structure. Although cleavage rates to the two-cell stage did not differ among groups, delayed cleavage was observed in the vanadate-treated group. The blastocyst formation rate was significantly reduced by vanadate treatment compared with caffeine-treated and non-treated (control) NT groups (P<0.05). The apoptotic cell index of NT blastocysts was lower in the caffeine-treated group than in other groups (P<0.05). The methylation patterns were similar among NTs, but more hypermethylated DNA was observed at the four-cell stage of control and vanadate-treated NTs when compared with that in IVF embryos (P<0.05). Thus, the nuclear remodelling type controlled by caffeine or vanadate treatment can affect in vitro development and the methylation status of NTs in relation to nuclear reprogramming. PMID- 18411412 TI - A new O6-alkylguanine-DNA alkyltransferase inhibitor associated with a nitrosourea (cystemustine) validates a strategy of melanoma-targeted therapy in murine B16 and human-resistant M4Beu melanoma xenograft models. AB - Chemoresistance to O(6)-alkylating agents is a major barrier to successful treatment of melanoma. It is mainly due to a DNA repair suicide protein, O(6) alkylguanine-DNA alkyltransferase (AGT). Although AGT inactivation is a powerful clinical strategy for restoring tumor chemosensitivity, it was limited by increased toxicity to nontumoral cells resulting from a lack of tumor selectivity. Achieving enhanced chemosensitization via AGT inhibition preferably in the tumor should protect normal tissue. To this end, we have developed a strategy to target AGT inhibitors. In this study, we tested a new potential melanoma-directed AGT inhibitor [2-amino-6-(4-iodobenzyloxy)-9-[4-(diethylamino) ethylcarbamoylbenzyl] purine; IBgBZ] designed as a conjugate of O(6)-(4 iododbenzyl)guanine (IBg) as the AGT inactivator and a N,N diethylaminoethylenebenzamido (BZ) moiety as the carrier to the malignant melanocytes. IBgBZ demonstrated AGT inactivation ability and potentiation of O(6) alkylating agents (cystemustine, a chloroethylnitrosourea) in M4Beu highly chemoresistant human melanoma cells both in vitro and in tumor models. The biodisposition study on mice bearing B16 melanoma, the standard model for the evaluation of melanoma-directed agents, and the secondary ion mass spectrometry imaging confirmed the concentration of IBgBZ in the tumor and in particular in the intracytoplasmic melanosomes. These results validate the potential of IBgBZ as a new, more tumor-selective, AGT inhibitor in a strategy of melanoma-targeted therapy. PMID- 18411413 TI - The effect of subchronic administration of 7-(4-fluorobenzyloxy)-2,3-dimethyl-1 {[(1S,2S)-2-methylcyclopropyl]methyl}-1H-pyrrolo[2,3-d]pyridazine (CS-526), a novel acid pump antagonist, on gastric acid secretion and gastrin levels in rats. AB - In the present report, we evaluated the effect of the novel acid pump antagonist 7-(4-fluorobenzyloxy)-2,3-dimethyl-1-{[(1S,2S)-2-methylcyclopropyl]methyl}-1H pyrrolo[2,3-d]pyridazine (CS-526) and 2-[3-methyl-4-(2,2,2-trifluoro-ethoxy) pyridin-2-ylmethanesulfinyl]-1H-benzimidazole (lansoprazole) on rebound gastric acid secretion, using an intragastric dialysis membrane perfusion model and on the serum and antral gastrin level after a 14-day treatment in rats. The effect of CS-526 on gastric acid secretion was almost constant during the 14 days of treatment. After the 14-day treatment, gastric acid secretion had returned to pretreatment levels. However, CS-526 slightly increased and lansoprazole potently increased gastric acid secretion thereafter. In the posttreatment period, the influence on rebound gastric acid secretion by lansoprazole treatment was significant, but that by CS-526 was not. The serum gastrin concentration after the 14-day treatment with CS-526 did not increase significantly, even at 100 mg/kg/day. On the other hand, lansoprazole at 100 mg/kg/day significantly elevated the serum gastrin concentration. After the 14-day treatment with CS-526 at 100 mg/kg/day, the antral gastrin content significantly increased. Lansoprazole at the doses of 30 and 100 mg/kg/day also significantly increased the antral gastrin content after the 14-day treatment. The elevation of the serum gastrin level after the lansoprazole treatment was suppressed by the concomitant administration of CS-526. In conclusion, CS-526 has a potent antisecretory effect on gastric acid secretion without rebound gastric hypersecretion. Moreover, CS 526 had minimal effects on the serum and antral gastrin elevation. It is suggested that these effects on gastric acid secretion and serum gastrin after subchronic treatment with CS-526 would be beneficial in clinical use. PMID- 18411414 TI - A concept analysis of cultural sensitivity. AB - Cultural sensitivity is used ubiquitously, yet different meanings are constructed. An improved understanding of the term as described within health care and in general would enhance nurses' understanding and communication with professionals and clients. To uncover the current meaning of cultural sensitivity, a concept analysis was performed. Findings included the attributes of knowledge, consideration, understanding, respect, and tailoring. Necessary antecedents were diversity, awareness, and an encounter. The consequences were effective communication, effective intervention, and satisfaction. A definition of cultural sensitivity was proposed. Providers may benefit from recognizing and addressing the identified antecedents and attributes to obtain the positive consequences of employing cultural sensitivity. PMID- 18411415 TI - Placenta growth factor augments endothelin-1 and endothelin-B receptor expression via hypoxia-inducible factor-1 alpha. AB - Pulmonary hypertension (PHT) develops in sickle cell disease (SCD) and is associated with high mortality. We previously showed that erythroid cells produce placenta growth factor (PlGF), which activates monocytes to induce proinflammatory cytochemokines, contributing to the baseline inflammation and severity in SCD. In this study, we observed that PlGF increased expression of endothelin-1 (ET-1) and endothelin-B receptor (ET-BR) from human pulmonary microvascular endothelial cells (HPMVECs) and monocytes, respectively. PlGF mediated ET-1 and ET-BR expression occurred via activation of PI-3 kinase, reactive oxygen species and hypoxia inducible factor-1 alpha (HIF-1 alpha). PlGF increased binding of HIF-1 alpha to the ET-1 and ET-BR promoters; this effect was abrogated with mutation of hypoxia response elements in the promoter regions and HIF-1 alpha siRNA and confirmed by chromatin immunoprecipitation analysis. Furthermore, PlGF-mediated ET-1 release from HPMVECs and ET-BR expression in monocytes creates a PlGF-ET-1-ET-BR loop, leading to increased expression of MCP 1 and IL-8. Our studies show that PlGF-induced expression of the potent vasoconstrictor ET-1 and its cognate ET-BR receptor occur via activation of HIF-1 alpha, independent of hypoxia. PlGF levels are intrinsically elevated from the increased red cell turnover in SCD and in other chronic anemia (eg, thalassemia) and may contribute to inflammation and PHT seen in these diseases. PMID- 18411416 TI - NOTCH1 extracellular juxtamembrane expansion mutations in T-ALL. AB - Heterodimerization domain (HD) mutations in NOTCH1 induce ligand-independent activation of the receptor and contribute to the pathogenesis of one-third of human T-cell lymphoblastic leukemias (T-ALLs). Here we report a novel class of activating mutations in NOTCH1 leading to aberrant activation of NOTCH1 signaling in T-cell lymphoblasts. These so-called juxtamembrane expansion (JME) alleles consist of internal duplication insertions in the vicinity of exon 28 of the NOTCH1 gene encoding the extracellular juxtamembrane region of the receptor. Notably, structure-function analysis of leukemia-derived and synthetic JME mutants demonstrated that the aberrant activation of NOTCH1 signaling is dependent on the number of residues introduced in the extracellular juxtamembrane region of the receptor and not on the specific amino acid sequence of these insertions. JME NOTCH1 mutants are effectively blocked by gamma-secretase inhibitors and require an intact metalloprotease cleavage site for activation. Overall, these results show a novel mechanism of NOTCH1 activation in T-ALL and provide further insight on the mechanisms that control the activation of NOTCH1 signaling. PMID- 18411417 TI - Extracorporeal photopheresis reverses experimental graft-versus-host disease through regulatory T cells. AB - Extracorporeal photopheresis (ECP), a technique that exposes isolated white blood cells to photoactivatable 8-methoxypsoralen and ultraviolet A radiation, is used clinically to treat cutaneous T-cell lymphoma and immune-mediated diseases such as graft-versus-host disease (GVHD). ECP is thought to control these diseases in part through direct induction of lymphocyte apoptosis, but its effects on the immune system beyond apoptosis remain poorly characterized. We have developed a novel method for incorporating ECP treatment into well-established and clinically relevant murine models of GVHD to examine its effects during an ongoing immune response. We demonstrate that the transfer of cells treated with ECP reverses established GVHD by increasing donor regulatory T cells and indirectly reducing the number of donor effector lymphocytes that themselves had never been exposed to psoralen and ultraviolet A radiation. PMID- 18411418 TI - Long-term results of the fludarabine, cyclophosphamide, and rituximab regimen as initial therapy of chronic lymphocytic leukemia. AB - Early results of the fludarabine, cyclophosphamide, and rituximab (FCR) regimen in 224 patients showed that it was highly active as initial therapy of chronic lymphocytic leukemia. In this report, we present the final results of all 300 study patients at a median follow up of 6 years. The overall response rate was 95%, with complete remission in 72%, nodular partial remission in 10%, partial remission due to cytopenia in 7%, and partial remission due to residual disease in 6%. Two patients (< 1%) died within 3 months of starting therapy. Six-year overall and failure-free survival were 77% and 51%, respectively. Median time to progression was 80 months. Pretreatment characteristics independently associated with inferior response were age 70 years or older, beta2-microglobulin twice the upper limit of normal (2N) or more, white cell count 150 x 10(9)/L or more, abnormal chromosome 17, and lactate dehydrogenase 2N or more. No pretreatment characteristic was independently associated with decreased complete remission duration. The risk of late infection was 10% and 4% for the first and second years of remission, respectively, and less than 1.5% per year for the third year onward. In a multivariate analysis of patients receiving fludarabine-based therapy at our center, FCR therapy emerged as the strongest independent determinant of survival. PMID- 18411419 TI - Eight-year experience with allogeneic stem cell transplantation for relapsed follicular lymphoma after nonmyeloablative conditioning with fludarabine, cyclophosphamide, and rituximab. AB - Nonmyeloablative stem cell transplantation in patients with follicular lymphoma has been designed to exploit the graft-versus-lymphoma immunity. The long-term effectiveness and toxicity of this strategy, however, is unknown. In this prospective study, we analyzed our 8-year experience. Patients received a conditioning regimen of fludarabine (30 mg/m(2) daily for 3 days), cyclophosphamide (750 mg/m(2) daily for 3 days), and rituximab (375 mg/m(2) for 1 day plus 1000 mg/m(2) for 3 days). They were then given an infusion of human leukocyte antigen-matched hematopoietic cells from related (n = 45) or unrelated donors (n = 2). Tacrolimus and methotrexate were used for graft-versus-host disease (GVHD) prophylaxis. Forty-seven patients were included. All patients experienced complete remission, with only 2 relapses. With a median follow-up time of 60 months (range, 19-94), the estimated survival and progression-free survival rates were 85% and 83%, respectively. All 18 patients who were tested and had evidence of JH/bcl-2 fusion transcripts in the bone marrow at study entry experienced continuous molecular remission. The incidence of grade 2-IV acute GVHD was 11%. Only 5 patients were still undergoing immunosuppressive therapy at the time of last follow-up. We believe that the described results are a step forward toward developing a curative strategy for recurrent follicular lymphoma. PMID- 18411420 TI - The proteosomal degradation of fusion proteins cannot be predicted from the proteosome susceptibility of their individual components. AB - It is assumed that the proteosome-processing characteristics of fusion constructs can be predicted from the sum of the proteosome sensitivity of their components. In the present study, we observed that a fusion construct consisting of proteosome-degradable proteins does not necessarily result in a proteosome degradable chimera. Conversely, fusion of proteosome-resistant proteins may result in a proteosome-degradable composite. We previously demonstrated that conserved influenza proteins can be unified into a single fusion antigen that is protective, and that vaccination with combinations of proteosome-resistant and proteosome-degradable antigens resulted in an augmented T-cell response. In the present study we constructed proteosome-degradable mutants of conserved influenza proteins NP, M1, NS1, and M2. These were then fused into multipartite proteins in different positions. The stability and degradation profiles of these fusion constructs were demonstrated to depend on the relative position of the individual proteins within the chimeric molecule. Combining unstable sequences of either NP and M1 or NS1 and M2 resulted in either rapidly proteosome degraded or proteosome resistant bipartite fusion mutants. However, further unification of the proteosome-degradable forms into a single four-partite fusion molecule resulted in relatively stable chimeric proteins. Conversely, the addition of proteosome resistant wild-type M2 to proteosome-resistant NP-M1-NS1 fusion protein lead to the decreased stability of the resulting four-partite multigene products, which in one case was clearly proteosome dependent. Additionally, a highly destabilized form of M1 failed to destabilize the wild-type NP. Collectively, we did not observe any additive effect leading to proteosomal degradation/nondegradation of a multigene construct. PMID- 18411421 TI - Using structural analysis to generate parasite-selective monoclonal antibodies. AB - Diagnosis of eukaryotic parasitic infection using antibody-based tests such as ELISAs (enzyme-linked immunosorbent assays) is often problematic because of the need to differentiate between homologous host and pathogen proteins and to ensure that antibodies raised against a peptide will also bind to the peptide in the context of its three-dimensional protein structure. Filariasis caused by the nematode, Brugia malayi, is an important worldwide tropical disease in which parasites disappear from the bloodstream during daylight hours, thus hampering standard microscopic diagnostic methods. To address this problem, a structural approach was used to develop monoclonal antibodies (mAbs) that detect asparaginyl tRNA synthetase (AsnRS) secreted from B. malayi. B. malayi and human AsnRS amino acid sequences were aligned to identify regions that are relatively unconserved, and a 1.9 A crystallographic structure of B. malayi AsnRS was used to identify peptidyl regions that are surface accessible and available for antibody binding. Sequery and SSA (Superpositional Structural Analysis) software was used to analyze which of these peptides was most likely to maintain its native conformation as a synthetic peptide, and its predicted helical structure was confirmed by NMR. A 22-residue peptide was synthesized to produce murine mAbs. Four IgG(1) mAbs were identified that recognized the synthetic peptide and the full-length parasite AsnRS, but not human AsnRS. The specificity and affinity of mAbs was confirmed by Western blot, immunohistochemistry, surface plasmon resonance, and enzyme inhibition assays. These results support the success of structural modeling to choose peptides for raising selective antibodies that bind to the native protein. PMID- 18411423 TI - Cholinesterase inhibitors for delirium: what is the evidence? PMID- 18411424 TI - Time to defibrillation after in-hospital cardiac arrest. PMID- 18411422 TI - Conformational change upon ligand binding and dynamics of the PDZ domain from leukemia-associated Rho guanine nucleotide exchange factor. AB - Leukemia-associated Rho guanine nucleotide exchange factor (LARG) is a RhoA specific guanine nucleotide exchange factor (GEF) that can activate RhoA. The PDZ (PSD-95/Disc-large/ZO-1 homology) domain of LARG interacts with membrane receptors, which can relay extracellular signals to RhoA signal transduction pathways. Until now there is no structural and dynamic information about these interactions. Here we report the NMR structures of the LARG PDZ in the apo form and in complex with the plexin-B1 C-terminal octapeptide. Unobservable resonances of the residues in betaB/betaC and betaE/alphaB loops in apo state were observed in the complex state. A distinct region of the binding groove in the LARG PDZ was found to undergo conformational change compared with other PDZs. Analysis of the (15)N relaxation data using reduced spectral density mapping shows that the apo LARG PDZ (especially its ligand-binding groove) is flexible and exhibits internal motions on both picosecond to nanosecond and microsecond to millisecond timescales. Mutagenesis and thermodynamic studies indicate that the conformation of the betaB/betaC and betaE/alphaB loops affects the PDZ-peptide interaction. It is suggested that the conformational flexibility could facilitate the change of structures upon ligand binding. PMID- 18411425 TI - Time to defibrillation after in-hospital cardiac arrest. PMID- 18411426 TI - Time to defibrillation after in-hospital cardiac arrest. PMID- 18411427 TI - Time to defibrillation after in-hospital cardiac arrest. PMID- 18411428 TI - Breast-cancer stromal cells with TP53 mutations. PMID- 18411429 TI - Breast-cancer stromal cells with TP53 mutations. PMID- 18411430 TI - Breast-cancer stromal cells with TP53 mutations. PMID- 18411431 TI - Paclitaxel plus bevacizumab for metastatic breast cancer. PMID- 18411432 TI - Expansion of the health workforce and the HIV Epidemic. PMID- 18411433 TI - Restricted diffusion on MR imaging of an acute cerebral lesion. PMID- 18411434 TI - Advising about return to work is a core activity for most occupational physicians. PMID- 18411435 TI - Bibliography. Current world literature. Biliary tract. PMID- 18411436 TI - Peru makes progress on maternal health. PMID- 18411437 TI - Community involvement saves newborn infants in India. PMID- 18411438 TI - [Secondary correction of nasolabial-complex and angle of mouth after insufficient dynamic temporal muscle transfer]. PMID- 18411439 TI - Ultraviolet-A and -B differentially modify the tyrosine-kinase profile of human keratinocytes and induce the expression of Arg+. AB - To investigate the expression profile of protein tyrosine kinases (PTKs) in normal human epidermal keratinocytes (NHEK) in response to UVA and UVB we employed a reversed transcriptase polymerase chain reaction (PCR) approach using degenerate primers derived from the conserved catalytic domain of PTKs. Quantitative real-time PCR with specific primers was used to confirm the influence of UV on the expression of the identified PTKs. Arg (Abelson-related gene, Abl2) was the PTK with the highest prevalence (30% of all PTKs) and UVA led to a further induction of Arg expression reaching nine-fold mRNA baseline expression at 17 h after irradiation. UVB was followed by an initial downregulation and a subsequent increase in Arg mRNA reaching five-fold baseline levels after 24 h. We conclude that UVA and UVB differentially modify the expression of PTKs in NHEK, and that Arg appears to have a major role in the response of keratinocytes to UV. These results provide a basis for further studies of PTK in UV-induced signaling that regulates protective responses, cell growth and carcinogenesis in the skin. PMID- 18411441 TI - [The prevalence and progression of the stomach and duodenum ulcer diseases among the employees of the Motor License and Inspection Department of Tatarstan]. PMID- 18411440 TI - [Society against heartburn]. PMID- 18411442 TI - [The role of tumor necrosis factors (TNF-alpha) in patients with chronic obstructive lung disease in combination with stomach and duodenum ulcer diseases]. PMID- 18411443 TI - [Characteristics of gastroesophageal reflux disease in elderly people]. PMID- 18411444 TI - [Problems of resistance during treatment of chronic hepapitis B with nucleoside and nucleotide analogues: diagnostics, prophylactics and treatment]. PMID- 18411445 TI - [Ulcer disease: critical analysis of contemporary conceptions of pathogenesis]. PMID- 18411446 TI - [Column and upper limb muscoloskeletal disorders in drivers helpers]. AB - The past two decades have witnessed a sharp rise in the incidence of work-related musculoskeletal disorders (WMSD) off the upper extremities and low back pain. The high frequency of these Pathologies and the estimated costs induced a large number of searcher to indentify causes etiological and preventive measure. Epidemiological surveys have inquired people mainly devoted to health clinic, where nurses and health assistants partner; are few data on the rescuers drivers and operators of emergencies. In the sample considered the 3.9% presented abnormalities in at least one section of the column, the 3.8% presented borne disorders of the cervical spine and backbone for the past 12 months. No major disturbances emerged dependents of the upper limbs, pain in the wrist, paraesthesia day and night; No subject also reported disturbances hands as evidenced from examination goal. The 2.3% reported having had an accident at work with prognosis of less than 30 days and 06% over 30 days. The results showed that, through prevention program it is possible to reduce musculoskeletal disorders in specific occupations such as drivers and operators of emergencies. PMID- 18411447 TI - [Estimate of the previous exposure to asbestos through mineralogical examination of the bronchoalveolar lavage (BAL) fluid in a population working in a sugar industry]. AB - In this study the past professional exposure and the health state of sugar refinery workers in Arezzo (Italy) are reviewed. The medical examinations, confirmed by chest CT, showed asbestos pleural thickening and one asbestosis case. Mineralogical analysis of bronchoalveolar lavage fluid (BALF) was carried out in this population to measure the pulmonary burden. Total fiber's concentration was not different in either subjects with or without asbestos related disease. Notably, comparison between total fiber's concentration in sugar refinery workers and subjects employed in other industries, revealed a lower professional exposure of the former with respect to shipyard workers and electric plants. In other groups of Italian sugar refinery workers, a concentration of asbestos fibers higher than in the studied population was found. Thus, the study highlights the necessity to develop standardized methodologies to carry out health intervention programmes in workers previously exposed to asbestos. PMID- 18411448 TI - [Shunting esophagocolonoplasty in the treatment of postburn cicatricial esophageal stricture]. AB - The prospective analysis of the results of treatment of postburn cicatricial esophageal stricture (PCES) in 338 (32%) patients, using shunting retrosternal esophagocolonoplasty (SRECP), constituting 76.1% of esophagoplasties performed, was conducted. Postoperative mortality was 4.7% (16 patients died). Early postoperative complications had occurred in 116 (34.5%) patients and the late--in 68 (24.9%). SRECP constitutes the method of first choice in patients, suffering tubular PCES and in formation of fistula, diverticulum and false ways. The standard variant of the procedure includes the performance of wide one-raw esophago-colonoanastomosis in end-to-end fashion with isoperistaltic passage of transplant. Such a tactic owes significant advantages, such as the lower risk of specific severe early and late postoperative complications occurrence, and also secures the esophagoplasty functional adequacy. PMID- 18411449 TI - [The choice of the surgical tactic in the treatment of acute purulent-necrotizing pancreatitis]. AB - Experience of treatment of 70 patients, suffering postnecrotic complications of an acute pancreatits, was presented. The method of three-dimension surgical reconstruction of purulent-necrotic focus, basing on the ultrasound investigation data analysis, was proposed. Three groups of patients were delineated accordance with pancreatic necrotic affection spread. The infectioning criterion, depending on the necrotic focus volume, was elaborated. Indications for surgical treatment were strictly directed on the basis of focus infectioning, it had envisaged the performance of programmed staged miniinvasive operative interventions. PMID- 18411450 TI - [Optimization of the program of complex perioperative treatment of patients with advanced purulent peritonitis]. AB - Comparative estimation of a standard and optimized curative programs, based on investigation of the systemic inflammation markers, the endogenic toxemia severity and intraabdominal pressure dynamics in 137 patients, suffering purulent peritonitis, was studied up. The efficacy of optimized program of complex treatment, application of which have had promoted the metabolic disorders elimination and the complicated postoperative course rate essential reduction, was suggested. PMID- 18411451 TI - [Late dysfunction of renal allotransplant: terminology, pathogenesis, morphological diagnosis]. PMID- 18411452 TI - [A knife wounding of the abdominal cavity organs with damage of the vena cava inferior, intestine, ureter]. PMID- 18411453 TI - [Surgical treatment of pulmonary cancer of the lung apex with subclavicular artery prosthesis]. PMID- 18411454 TI - Meth use among HIV-infected MSM poses multiple physical & behavioral problems. Scientists are closer to understanding why. PMID- 18411455 TI - Circumcision does not affect women's STD risk. PMID- 18411456 TI - New model of adherence teaches IMB skills. Motivation often is the biggest barrier. AB - Researchers suggest combining motivation assessment with intervention aimed at providing information, motivation, and behavioral skills. PMID- 18411457 TI - Veterans have higher rate of non-AIDS-related cancers. Clinicians should screen for certain cancers. AB - HIV-positive veterans are more likely to develop cancer than uninfected veterans, study shows. PMID- 18411458 TI - FDA notifications. FDA provides more info in response to Viracept recall. PMID- 18411459 TI - Microbicide researchers are focusing on adherence strategies as setbacks mount. IAS conference held uninspiring news. PMID- 18411460 TI - Expert offers look at issues in microbicide research. Here is what needs to be overcome. PMID- 18411461 TI - Bioethics program prepares Africans to work in HIV care. PMID- 18411462 TI - FDA notifications. Maraviroc approved as a CCR5 co-receptor antagonist. PMID- 18411463 TI - FDA notifications. FDA grants tentative approval of efavirenz. PMID- 18411464 TI - Hormonal contraception and HIV risk: a review. PMID- 18411465 TI - Lactic acidosis in Africans receiving HAART. PMID- 18411466 TI - "I don't want to grow up." Transitioning HIV-infected adolescents to adult care. PMID- 18411467 TI - Six rapid HIV tests are available. How do they compare? PMID- 18411468 TI - Update: the state of HIV research at MCLNO's HOP Clinic. PMID- 18411469 TI - Medication adherence: what are the barriers and facilitators? PMID- 18411470 TI - Update: the oral lesions associated with immune suppression. PMID- 18411471 TI - An examination of disclosure issues for HIV-infected adolescents and young adults. PMID- 18411472 TI - Progress notes: a New Orleans HIV clinic 22 months post-Katrina. PMID- 18411473 TI - Pain management in HIV/AIDS: an update for clinicians. PMID- 18411474 TI - Reality demands change: people over age 50 living with HIV continues to increase. PMID- 18411475 TI - Depression and distress in older HIV+ adults. PMID- 18411476 TI - Cardiovascular disease and HIV. PMID- 18411477 TI - Taking care of each other. AB - The first decade of AIDS was defined by death and activism, the second by medicines and hope. How can we rally again to make the third decade one of successful aging? PMID- 18411478 TI - CDC is working on action plan to target HIV epidemic among US Latino population. Recent MMWR highlights concern. PMID- 18411479 TI - Latinos and HIV epidemic. Intervention "effective program" for Latino youth. Intervention works equally well with boys and girls. PMID- 18411480 TI - IDU researchers face myriad of ethical dilemmas. IRBs may raise the wrong concerns. PMID- 18411481 TI - Effects of antiretroviral agents on lipid panels. PMID- 18411482 TI - Screening for abacavir hypersensitivity. PMID- 18411483 TI - FDA notifications. Revised guidelines for pregnant women. PMID- 18411484 TI - FDA notifications. FDA draft guidance is about new drug development. PMID- 18411485 TI - FDA notifications. Accelerated approval for raltegravir tablets. PMID- 18411486 TI - FDA notifications. Tentative approval to Aptivus for combination ART. PMID- 18411487 TI - Flu shot reminder. AB - A good time to get the annual shots is October or November, before the flu season begins. PMID- 18411488 TI - New kind of antiretroviral, KP-1461; clinical trial recruiting. Interview with Stephen Becker, M.D. AB - KP-1461, an experimental HIV drug already in a phase II trial, works so differently from other antiretrovirals that at first glance it looked like science fiction, and we found it hard to take seriously as a current possibility today. In fact this drug is highly credible, and based on elegant science that goes back at least 25 years. KP-1461 is the only antiretroviral in human use or testing that can eradicate HIV from laboratory cell cultures. No one knows how it will work in people -- but we might know by the second quarter of 2008, when the current phase II trial could be complete. AIDS Treatment News interviewed Dr. Stephen Becker, a leading AIDS physician and researcher who is vice president of clinical development at Koronis Pharmaceuticals, in Seattle, Washington. PMID- 18411490 TI - T-wave alterans magnitude, T-wave amplitude, and heart rate. Commentary on "Characterization of T wave alterans with ambulatory electrocardiography". PMID- 18411489 TI - Elevated serum polybrominated diphenyl ethers and thyroid-stimulating hormone associated with lymphocytic micronuclei in Chinese workers from an E-waste dismantling site. AB - In this study, we recruited 49 subjects from one village close to an electronic waste (e-waste) site (exposed group) and another located 50 km away from the e waste site (control group). We found that serum levels of polybrominated diphenyl ethers (median PBDEs, 382 ng/g lipid weight; range, 77-8452 ng/g lipid weight) and thyroid-stimulating hormone (median TSH, 1.79 microIU/mL; range, 0.38-9.03 microIU/mL) and frequencies of micro-nucleated binucleated cells (MNed BNC; median, 5% per hundred; range, 0-96% per hundred) were significantly higher in the exposed group than in the control group (158 ng/g, range of 18-436 ng/g, and p < 0.05; 1.15 microIU/mL, range of 0.48-2.09, and p < 0.01; and 0% per hundred, range of 0-5% per hundred, and p < 0.01, respectively). A history of working with e-waste was significantly associated with increased MNed BNC frequencies (odds ratio (OR), 38.85; 95% confidence interval (CI) = 1-1358.71, p = 0.044), independent of years of local residence, a perceived risk factor. However, there was no association between PBDEs exposure and oxidative DNA damage. Therefore, the exposure to PBDEs at the e-waste site may have an effect on the levels of TSH and genetoxic damage among these workers, but this needs to be validated in large studies. PMID- 18411491 TI - Health-related quality of life in rheumatoid arthritis. PMID- 18411492 TI - Evidence from many countries links higher income to better health status. Introduction. PMID- 18411493 TI - [Workplace mobbing]. AB - Workplace mobbing is a hostile and unethical communication, systematically aimed from one or more individuals towards mostly one individual, who are forced into a helpless position and are held in it by constant bullying. This article describes some of the most important characteristics of mobbing: offensive behaviour, organizational and non-organizational causes of this behaviour, the victim and the consequences. Modern business environment is complex, dynamic, volatile, and requires better ability to adjust. Constant changes are a part of organizational reality, but they also produce an ideal environment for all kinds of conflicts. Conflicts are inevitable in every organization, but the task of its management is to identify them and resolve before they affect the workforce, productivity and costs. The idea is to avert psychological abuse and aberrant behaviour such as mobbing which that may cause physical and mental disorders. Mobbing is a problem of the modern society; as a violation of human rights it is relatively new and unrecognised in Croatia. Abuse is mostly psychological: it affects the victim's health and life, quality of work, productivity, profitability, and may lead to significant economic losses in the community. Mobbing can be averted by joint forces that would involve employee and management, medical and legal professionals, and even community as a whole. The more an organization pursues excellence based on trust and business ethics, the higher the probability that mobbing will be averted or stopped. PMID- 18411494 TI - Screening newborns: just the beginning. Newborn screening and tracking diagnosed children into adulthood varies among states. PMID- 18411495 TI - License to serve: a model law could make it easier for medical volunteers to respond immediately during emergencies. PMID- 18411496 TI - Consumer food subsidies in India: proposals for reform. PMID- 18411497 TI - Richer and taller: stature and living standards in China, 1979-1995. PMID- 18411498 TI - Voting by older adults with cognitive impairments. AB - This presidential election year reminds us of the importance of each vote and of the integrity of the electoral process. Recent elections have been decided by very narrow margins. In this context, the voting rights and capacity of persons with dementia warrant attention. About 4.5 million Americans currently live with dementia. Whether these citizens should vote raises a host of ethical, legal, and practical issues. At what point does someone lose the capacity to vote, and who decides? What kinds of assistance should these voters get, and who should provide it? And how can the voting rights of residents in long-term care facilities be protected? PMID- 18411499 TI - Use of GeneSearch breast lymph node assay to detect sentinel node metastases in early stage breast cancer. PMID- 18411500 TI - Genetic testing for long QT syndrome. PMID- 18411501 TI - Computer-assisted navigation for total knee arthroplasty. PMID- 18411502 TI - Off-label uses of sorafenib and sunitinib. PMID- 18411503 TI - Artificial intervertebral disc arthroplasty for treatment of degenerative disc disease of the cervical spine. PMID- 18411505 TI - [Abstracts of the 11th Atherosclerosis Congress sponsored by the Czech Atherosclerosis Society in cooperation with IAS. Spindleruv Mlyn, 6-8 December 2007]. PMID- 18411504 TI - Report of the Poison Control Centre for the period 1 January - 31 December 2007. PMID- 18411506 TI - [Communication in life science research: Research communication activity in genome studies]. PMID- 18411508 TI - [Thiosubstrates of different cholinesterases]. AB - Review of the own and literature data on substrate specificity with use of thiosubstrates for cholinesterases of various species. Dependence of cholinesteratic hydrolysis parameters on various elements of their structure is considered: the acyl part, alkyl "bridge" between ester atom and onion group, and ammonium grouping of molecule of 44 thioesters. A comparative enzymological analysis of the substrate specificity is performed with use of thiocholine esters of acetic, propionic, and butyric acids for 40 cholinesterase preparations of mammals, insects, molluscs, and plants. PMID- 18411507 TI - [Signal pathway of insulin and insulin-like growth factor 1 (IGF-1) as a potential regulator of lifespan]. AB - The experimental material accumulated for two decades allows concluding that regulation of lifespan has hormonal control based on the evolutionary conservative insulin/IGF-1 receptor signal pathway. Data obtained on the commonly accepted models of longevity - nematode Caenorhabditis elegans, Drosophila Drosophila melanogaster, and rodents - demonstrate that reduction of the insulin/IGF- 1 signal pathway leads to an increase of the lifespan. There is shown involvement of the longevity mechanism of a large group of genes whose products perform control of metabolism, alimentary behavior, reproduction, resistance to oxidative stress. Discussed in this review are current concepts of the insulin/IGF-1 signal system as a regulatory "longevity module" and of its possible role in prolongation of life in the higher vertebrates, including human. PMID- 18411509 TI - [Use of properties and regulation peculiarities of enzymes of glycogenolysis in fish skeletal muscle depending on peculiarities of motor activity of species]. AB - Levels of activity, properties, and peculiarities of activation of glycogen phosphorylase (GP; EC 2.4.1.1) and glycogen phosphorylase kinase (GPK; EC 2.7.1.38) were studied in the white skeletal muscle of fish differing in motor behavior. No differences in the GP and GPK activity levels were revealed in laskir Diplodus annularis (L.), horse mackerel Trachurus mediterraneus ponticus, salmon Salmo trutta morphario, scorpena Scorpaena porcus, Scophtalnus maeoticus, and carp Cyprinus carpio; however, properties of the isolated enzymes and peculiarities of formation of their activated forms during swimming in a hydrodynamic tube are determined by functional peculiarities of the muscle tissue and are associated with the motor activity character of the species. In fish capable for the spurt type of swimming (scorpena, salmon) the more rapid ion regulation plays the predominant role. In other species, the glycogenolysis hormonal regulation leading to a change of the GPK activity index has been found. PMID- 18411510 TI - [PC12 cells transfected with human mutant gene causing one of Alzheimer's disease forms have a high sensitivity to oxidative stress]. AB - Used in this work are PC12 cells transfected with human gene expressing amyloid precursor protein of beta-peptide and carrying the so-called "Swedish mutation" leading to the appearance of one Alzheimer's disease family forms. It has been shown that the PC12 cells transfected with this mutant gene, at action of various hydrogen peroxide concentrations, die to the significant greater degree than the used for comparison PC12 cells transfected with analogous human gene of the wild type or than vector-transfected cells. It has been found that ganglioside GM1 at micro- or nanomolar concentrations is able to increase viability of the PC12 cells transfected with the mutant gene causing a significant accumulation of endogenous amyloid beta-peptide. The obtained data confirm an important role of oxidative stress in injury and death of brain nerve cells in Alzheimer's disease. PMID- 18411511 TI - [Mathematical model for cooperation of ion pump, symport, and antiport in epithelial cells]. AB - Transcellular transport in epithelial cells plays an important role in providing such physiological functions as excretion of cytotoxic substances or reabsorption of metabolites useful for the body life activity. These functions have been shown to be performed by the mechanisms - symport, antiport, ion pumps, and channels - that often function cooperatively. Kinetic models of the substrate transport with the aid of the above mechanisms are widely described in the literature. Much less attention is paid to modeling of cooperative activity of transporters that have different transport mechanisms. In this work we propose a mathematical model for flux coupling of three transporters - the ion pump, symporter, and antiporter as well as of two substrates, one of which (A) can be transported simultaneously by the symport and antiport mechanisms, while the other (B) - only by the latter mechanisms. Analysis of the model has shown that for the pair of substrates (A and B) the flux coupling becomes possible if the following conditions are met: (1) the substrate A flux into the internal cell volume using the symport mechanism is to exceed its antiporter-realized flux in the opposite direction; (2) probability of reorientation from one side of membrane to the other side for the antiporter loaded with the substrate is to be essentially higher than that for empty transporter. The proposed model can be used for comparing efficiency both of excretion and of reabsorption of cell metabolites in representatives of different taxa. PMID- 18411512 TI - [Peculiarities of Ca2+ regulation of functional activity of myocardium of frog Rana temporaria]. AB - To elucidate role of intra- and extracellular Ca2+ in regulation of rhythm and strength of frog heart contractions, there were studied ECC and isometric contraction of myocardium preparations in response to verapamil, adrenaline, and blockers of alpha- and beta-adrenoreceptors. It has been shown that after an intramuscular injection of verapamil (6 mg/kg), bradycardia develops, the heart rate (HR) decreasing by 50-70 %. Further, the cardiac arrest occurred; however, administration to the animals of adrenaline (100 mg/kg) restored the cardiac rhythm for a short while. After an intramuscular injection of adrenaline at doses of 0.1-10 mg/kg, no essential changes were observed in the potential action amplitude and HR; an increase of the administered adrenalin concentration to 100 mg/kg was not accompanied by the cardiac rhythm stimulation, as this takes place in homoiothermal animals and human; on the contrary, an essential HR deceleration was revealed. Phentolamine (5 mg/kg) gradually decelerated HR rhythm by 32-45 %. The potential amplitude changed insignificantly. A subsequent intracardiac injection of adrenaline (100 mg/kg) on the background of block of alpha adrenoreceptors produced acceleration of the rhythm (by 13-21%) and fall of the electrogram amplitude. These results can indicate that in the frog heart, phentolamine interacts predominanty with alpha-adrenoreceptors. An intracardiac administration of propranolol (1 mg/kg) to frogs promoted inhibition of beta adrenergic receptors and produced a gradual cardiac rhythm deceleration. In experiments on assessment of verapamil effect on the character of contractions this preparation at a concentration of 150 microM was established to produce a significant dosedependent decrease of the contraction strength. A rise of verapamil concentration in the sample to 200 microM led to a decrease of the amplitude, on average, by 68-70 % and in individual preparations -- by 80-85 %; however, administration into the sample of adrenaline (10 microM) restored the cardiac contraction strength. Adrenaline (1 nM--100 microM) increased markedly the contraction amplitude. Phentolamine (10 microM) did not inhibit transmission of contractile signal to cardiomyocytes; this was manifested in that the contraction amplitude after addition of adrenaline (10 microM) into the sample was approximately the same as in the sample containing no phentolamine. Propranolol (10 microM) eliminated the stimulatory action of adrenaline (10 microM). The results of these experiments indicate that in the frog ventricular cardiomyocytes the main adrenaline acceptors are beta-adrenoreceptors. PMID- 18411513 TI - [Evolutionary-physiological aspects of adaptation of the Pacific salmon fry of the Oncorhynchus genus to migration to the sea water]. AB - A complex of adaptive changes occurring in the Pacific salmon fry in the process of migration to the sea is described, including behavior, ion content in carcasses, and morphological changes in Stannius bodies, gill epithelium, and nephron tubular epithelium. Participating in experiments with transfer from fresh water into a two-layer aquarium (the lower layer - sea water, the upper layer - fresh water) were smolts of chum salmon and underyearlings of masu salmon as well as the trachurus and leiurus forms of the three-spined stickleback Casterosteus aculeatus. All fish, regardless of their salt preference, at once after placement into the two-layer aquarium, occupied the sea water zone, at the very bottom of the aquarium. After 1 h, there started brief excursions of masu salmon and chum salmon to the upper, fresh water layer; however, both forms of the three-spined stickleback did not participate in these excursions. After 12 h, the chum salmon settled down in the lower, sea water layer, while the masu salmon - in the upper, fresh water layer. Both forms of the three-spined stickleback never left the sea water layer and felt quite comfortably on the aquarium bottom. It seems that the high tolerance of the both stickleback forms to wide salinity limits allows them to choose the convenient position regardless of the water salt composition. By analyzing the material obtained for three years (2001-2003) on structure and functions of the gill epithelium chloride cells (CC), we have come to the conclusion that the fresh water fry of two salmon species, chum and masu salmons, caught at the same time and practically in the same water reservoirs can be divided into three groups. The underyearlings of the masu salmon as a rule are characterized by the thickened epithelium of secondary gill lamellae, but by a very small number of CC. In smolts of chum salmon, on the contrary, the epithelium is sufficiently thin, but enriched in the CC that demonstrate an active structure in the very beginning of migration to sea. However, with approaching the sea (and with an increase of terms of migration) the CC activity drops, but their amount does not change. And only after migration to the sea the CC activity rises again, although their amount seems to remain unchanged. The described peculiarities of behavior and of the ion composition regulation in the migrating salmon fry confirm the hypothesis that the salmons evolutionized in fresh water, that the Oncorhynchus genus appeared in large spaces of saltish waters, such as the Japan Sea at the period of the early Pleistocene, and that learning of fry of the Oncorhynchus genus (for instance, of O. gorbuscha and O. keta) is the most specialized in the salmons migrating to the sea, whereas the fresh water species of chars (Salvelinus) and of trouts (Salmo) are more primitive. PMID- 18411514 TI - [Changes of the serotonergic system activity in fish tissues during an increase of water salinity]. AB - Changes of environment parameters produce essential effects on the serotonergic system activity in the body tissues. This paper describes study of the serotonergic system activity in various tissues of half-migratory fish - the Caspian roach Rutilus rutilus caspicus and the bream Abramis brama orientalis - caught in the fresh and brackish water. Using the method of indirect solid-phase ELISA test, the serotonergic system activity was assessed by determining in the studied fish tissues of the amount of serotonin-modulating anticonsolidation protein (SMAP) that is in the linear correlation with serotonin level. A significant change of the SMAP level in the Caspian roach and bream brain was found under effect on increased water salinity. Analysis of the SMAP amount in the bream liver did not reveal any differences in the animals caught in the brackish and fresh water. At the same time, in the bream gill there was a small rise of the SMAP content in the fish caught in the brackish water. The revealed in this experiment elevation of the SMAP amount in the Caspian roach and bream brain reflecting a corresponding increase of the serotonergic system activity indicates involvement of adaptive readjustments in the animal body. PMID- 18411515 TI - [Protein 70 kDa in control of sleep and thermoregulation]. AB - Studies of expression of molecular chaperones of the family of Heat Shock Proteins 70 kDa (HSP70) in the mouse and rat brain during sleep deprivation do not answer the question whether the HSP70 produce somnogenic effect. In the present work there are studied effects of exogenous Hsp70 that is known to be able to penetrate into living cells in vitro and to acquire properties of endogenous chaperone. Hsp70 was microinjected into the third brain ventricle of rats and pigeons at the beginning of the inactive period of the day when under natural conditions the sleep duration increases and the somato-visceral parameters decrease. Hsp70 was found to enhance this natural process and to produce an additional increase in the total time of slow-wave sleep, a more pronounced inhibition of the muscle contractive activity, and a deeper decrease in the brain temperature. A similarity in effects of Hsp70 in rats and pigeons was revealed. In both species the somnogenic effect of Hsp70 in is realized by activation of mechanisms of maintenance of in longer episodes of in slow-wave sleep. The hypothermic Hsp70 effect seems to be associated with a decrease in the muscle contractive activity level, rather than with an enhancement in peripheral vasodilation and with an increase of heat loss. A hypothesis is put forward that the neuroleptic effect of Hsp70 that includes the somnogenic, myorelaxing, and hypothermic effects is mediated by activation of GABAA receptors of the main inhibitory brain system. PMID- 18411516 TI - [Compensatory reaction during degeneration of arcuate nucleus dopaminergic neurons in rats]. AB - The study has been carried out to verify the authors' hypothesis that degeneration of dopaminergic (DA-ergic) neurons of the hypothalamic tuberoinfundibular system and concomitant development of hyperprolactinemia are accompanied by involvement of compensatory synthesis of dopamine (DA) by non dopaminergic neurons expressing single complementary enzymes of synthesis of this neurotransmitter. Degeneration of DA-ergic neurons was produced by a stereotaxic injection into the brain lateral ventricles of 6-hydroxydopamine (6-OHDA) - a specific neurotoxin of DA-ergic neurons. 14 and 45 days after the toxin administration there were determined concentration of prolactine in peripheral blood by methods of immunoenzyme and radioimmunological analyses as well as the DA amount in the arcuate nucleus by the method of highly efficient liquid chromatography with electrochemical detection. In a part of the animals, slices were prepared from the mediobasal hypothalamus (arcuate nucleus and medial eminence) and perfused with Krebs-Ringer medium; then the DA concentration was determined in the slices and in the incubation medium. 14 days after the neurotoxin administration there were revealed an increase of blood prolactine concentration and a decrease of DA concentration in the arcuate nucleus in vivo as well a decrease of the total DA amount in the slices and incubation medium in experiments in vitro. 45 days after the neurotoxin administration, all the above parameters returned to the normal level. This, the obtained data indicate that the hyperlactinemia and DA deficit appearing during degeneration of the arcuate nucleus DA-ergic neurons seem to be compensated due to an enhancement of DA synthesis by non-dopaminergic monoenzyme neurons of arctuate nucleus. PMID- 18411517 TI - [Effect of harmaline on firing pattern of rat cerebellar Purkinje cells in ontogenesis]. AB - In this work, responses of rat Purkinje cells to intraperitoneal administration of the hallucinogenic alkaloid harmaline (0.15 mg/kg) were studied in the course of ontogenesis. The experiments were carried out on Wistar rats of three age groups: rat pups (13-18 days), adult animals (2-7 months), and aged rats (25-36 months). In Purkinje cell firings, two types of electric reactions were revealed; they were similar in all age group of the animals. In cells with the 1st type of reactions, in response to the harmaline administration there was recorded a significant increase of frequency of complex spikes, accompanied by disappearance of simple spikes. In the activity of Purkinje cells of the 2nd type, the complex spike frequency also increased; however, the firing simple spikes were preserved, although with a decrease of their frequency as compared with norm. Essential changes of activity of the cerebellar Purkinje cells were found in the rat pups and aged animals in comparison with adult rats, which agrees well with immaturity of various cerebellar structures in the first case and with involutionary changes in the second case. PMID- 18411519 TI - [Postnatal development of the cerebellar Purkinje cell shape in guinea pig ontogenesis]. AB - In sagittal cerebellum sections, morphometrical study of cerebellum of mature born animals - guinea pigs - was performed using Nissl's procedure. A change of shape and volume of Purkinje cells and their nuclei in the course of the guinea pig postnatal ontogenesis was studied. It has been shown that both the growth process itself and the rate of formation of the definite form of Purkinje cells and of their nuclei in the course of ontogenesis proceeds non-uniformly. The most intensive growth of vertical and horizontal diameters of Purkinje cells and of their nuclei is observed during the 1st and 4th weeks of postnatal life. Especially rapid is an increase of horizontal diameters of Purkinje cells and of their nuclei, which impairs the ovoid-bear-like shape to the cerebellar Purkinje cells of adult guinea pigs. PMID- 18411518 TI - [Architectonics of the central nervous system in Acoela, Plathelminthes, and Rotifera]. AB - Based on the literature and own data, consecutive stages of development of the central nervous system (CNS) in the lower Bilateria are considered - separation of brain from parenchyma, formation of its own envelopes, and development of the stem and orthogonal nervous system. Results of histochemical (cholinergic and catecholaminergic) and immunocytochemical (5-HT- and FMRFamid immunoreactive) studies of the CNS in representatives of Acoela, free living and parasitizing Plathelminthes and Rotifera are considered. The comparative analysis makes it possible to describe development and complication of the initially primitive Bilateria plexus nervous system. A special attention will be paid to the Acoela phylogenesis, based on molecular-biology data and results of study of their nervous system. PMID- 18411520 TI - [Kinetics of positive inotropic reaction of the snail Helix pomatia heart to serotonin]. PMID- 18411521 TI - Computer-assisted self-interviews: a cost effectiveness analysis. AB - Computer-assisted self-interview (CASI) questionnaires are being used with increased frequency to deliver surveys that previously were administered via self administeredpaper-and-pencil questionnaires (SAQs). Although CASI may offer a number of advantages, an important consideration for researchers is the assessment modality's immediate and long-term costs. To facilitate researchers' choice between CASI and SAQ, this article provides theoretical cost models with specific parameters for comparing the costs for each assessment type. Utilizing these cost models, this study compared the cost effectiveness in a health behavior study in which both CASI (n = 100) and SAQ (n = 100) questionnaires were administered. Given the high initial costs, CASI was found to be less cost effective than SAQ for a single study. However, for studies with large sample sizes or when CASI software is to be used for multiple studies, CASI would be more cost effective and should be the assessment mode of choice. PMID- 18411522 TI - Computer assessment of interview data using latent semantic analysis. AB - Clinical interviews are a powerful method for assessing students' knowledge and conceptualdevelopment. However, the analysis of the resulting data is time consuming and can create a "bottleneck" in large-scale studies. This article demonstrates the utility of computational methods in supporting such an analysis. Thirty-four 7th-grade student explanations of the causes of Earth's seasons were assessed using latent semantic analysis (LSA). Analyses were performed on transcriptions of student responses during interviews administered, prior to (n = 21) and after (n = 13) receiving earth science instruction. An instrument that uses LSA technology was developed to identify misconceptions and assess conceptual change in students' thinking. Its accuracy, as determined by comparing its classifications to the independent coding performed by four human raters, reached 90%. Techniques for adapting LSA technology to support the analysis of interview data, as well as some limitations, are discussed. PMID- 18411523 TI - RAP: a computer program for exploring similarities in behavior sequences using random projections. AB - A computer program (RAP, for random projection) for exploring similarities between and within sequences of behavior is presented. Given a time window of a sequence, the program calculates a signature, a real-valued vector that is a random projection of the contents of the window (i.e., the codes occurring within it and their relative location, or onset and offset times) into an arbitrary K dimensional space. Then, given two different time windows from the same sequence or from different sequences, their similarity is computed as an inverse function of the Euclidean distance between their respective signatures. By defining moving (overlapped or not overlapped) windows along each sequence and calculating similarities between every pair of windows from the two sequences, a map of similarities or possible recurrent patterns is obtained; the RAP program represents them as gray-level lattices, which are displayed as mouse-sensitive images in an HTML file. Computation of similarities is based on the random projection method, as presented by Mannila and Seppanen (2001), for the analysis of sequences of events. The program reads sequence data files in Sequential Data Interchange Standard (SDIS) format (Bakeman Quera, 1992,1995a). PMID- 18411524 TI - Windows executable software for the progressive demasking task. AB - The progressive demasking (PDM) task is a perceptual identification task using degraded stimulus presentation procedures, but with the advantage of providing response time (RT) measures on each trial. Participants simply press a response button whenever they recognize a word. In this article, we present newly developed, easy-to-use freeware for PDM. Considering the difficulty of programming such a paradigm with standard experimental software, we developed a program based on high-level C libraries to be executed on Windows XP computers. The program execution is millisecond accurate forRT measurements and generates the fewest possible display duration errors. PMID- 18411525 TI - Computer programming for generating visual stimuli. AB - Critical to vision research is the generation of visual displays with precise control over stimulus metrics. Generating stimuli often requires adapting commercial software or developing specialized software for specific research applications. In order to facilitate this process, we give here an overview that allows nonexpert users to generate and customize stimuli for vision research. We first give a review of relevant hardware and software considerations, to allow the selection of display hardware, operating system, programming language, and graphics packages most appropriate for specific research applications. We then describe the framework of a generic computer program that can be adapted for use with a broad range of experimental applications. Stimuli are generated in the context of trial events, allowing the display of text messages, the monitoring of subject responses and reaction times, and the inclusion of contingency algorithms. This approach allows direct control and management of computer generated visual stimuli while utilizing the full capabilities of modern hardware and software systems. The flowchart and source code for the stimulus-generating program may be downloaded from www.psychonomic.org/archive. PMID- 18411526 TI - External validation of the computerized, group administrable adaptation of the "operation span task". AB - One of the most widely used tasks for measuring working memory capacity is the operation span task (OSPAN; Turner & Engle, 1989). This task has almost always been applied individually, and stimuli presentation is controlled by the experimenter. Recently, De Neys, d'Ydewalle, Schaeken, and Vos (2002) improved the administration procedure by designing an automated, group-administrable version of the task (GOSPAN). They found GOSPAN to be reliable, and they also provided evidence on its validity (a significant positive correlation between GOSPAN and OSPAN scores). However, an external test of GOSPAN validity is still lacking. In this work, we present such a validation for the automated version, when the task is administered both individually (Experiment 1) and to groups (Experiment 2). There are abundant previous data on the relation between working memory capacity and reading comprehension. In this work, this relation is studied using an automated OSPAN version to measure working memory capacity. Given that our results are similar to those found using the original OSPAN, our data support the external validity of the automated version of the task. We also tested the reliability of the task and found high internal consistency in both experiments. PMID- 18411527 TI - A graphical representation of the mediated effect. AB - Mediation analysis is widely used in the social sciences. Despite the popularity of mediation models, few researchers have used graphical methods, other than structural path diagrams, to represent their models. Plots of the mediated effect can help a researcher better understand the results of the analysis and convey these results to others. This article presents a method for creating and interpreting plots of the mediated effect for a variety of mediation models, including models with (1) a dichotomous independent variable, (2) a continuous independent variable, and (3) an interaction between an independent variable and the mediating variable. An empirical example is then presented to illustrate these plots. Sample code for creating plots of the mediated effect in R and SAS is also included, and may be downloaded from www.psychonomic.org/archive. PMID- 18411528 TI - Diffusion model analysis with MATLAB: a DMAT primer. AB - The Ratcliff diffusion model has proved to be a useful tool in reaction time analysis. However, its use has been limited by the practical difficulty of estimating the parameters. We present a software tool, the Diffusion Model Analysis Toolbox (DMAT), intended to make the Ratcliff diffusion model for reaction time and accuracy data more accessible to experimental psychologists. The tool takes the form of a MATLAB toolbox and can be freely downloaded from ppw.kuleuven.be/okp/dmatoolbox. Using the program does not require a background in mathematics, nor any advanced programming experience (but familiarity with MATLAB is useful). We demonstrate the basic use of DMAT with two examples. PMID- 18411529 TI - Examining mixed unknown distributions (mud). AB - A function, written in R, for testing whether the distribution of responses in one condition can be considered a combination of the distributions from two other conditions is described. The important aspect of this function is that it does not make any assumptions about the shape of the distributions. It is based o nthe Kolmogorov-Smirnov D statistic. The function also allows the user to test more specific and, hence, more statistically powerful hypotheses. One hypothesis, that the mixture does not capture the middle third of the distribution, is included as a built-in option, and code is provided so that other alternatives can easily be run. A power analysis reveals that the function is most likely to detect a difference between the combined conditions' distribution and the other distribution when the center of the other distribution is near the midpoint of the two original distributions. Critical p values are estimated for each set of distributions, using bootstrap methods. An example from human memory research, exploring the blending hypothesis of the misinformation effect, is used for illustrative purposes. PMID- 18411531 TI - WolStat: a new statistics package for the behavioral and social sciences. AB - WolStat differs in several important ways from other statistics packages. Data are entered using a text editor, as opposed to a data grid. WolStat performs all possible simple to simple simple simple simple main effects tests and multiple comparison tests for ANOVAs (independent groups, split plot, and randomized block factorial) with up to five factors. More nonparametric statistics can be performed than with most other statistics packages. Latin squares can be randomly generated. Uniformly distributed random integers (with or without replacement) or normally distributed random numbers can be generated. WolStat provides a tutorial and assistance for selecting orthogonal comparisons. WolStat also provides general and specific models for fixed, mixed, and random ANOVAs with one through five factors. PMID- 18411530 TI - Using statistical smoothing to estimate the psycholinguistic acceptability of novel phrases. AB - Native speakers of languages perceive differences in the acceptability of phrases even when those phrases are both grammatical and novel (previously unseen). We suggest that smoothing, a statistical technique used by natural language processing engineers, provides several candidate mechanisms for investigating this phenomenon. We describe the creation of a large data set of predictions from several smoothing algorithms about the acceptability of unseen grammatical phrases and a novel experimental method for the pairwise comparison of these models. We use this method to compare three smoothing methods and consider the results in light of the differences among the models. We argue that the data support the idea that similarity in this domain is best thought of as a form of asymmetric representational distortion and that the informational basis over which such estimates are made is broad, rather than narrow, as has been previously suggested. PMID- 18411532 TI - Robust principal components: a generalized variance perspective. AB - This article compares several methods for performing robust principal component analysis, two of which have not been considered in previous articles. The criterion here, unlike that of extant articles aimed at comparing methods, is how well a method maximizes a robust version of the generalized variance of the projected data. This is in contrast to maximizing some measure of scatter associated with the marginal distributions of the projected scores, which does not take into account the overall structure of the projected data. Included are comparisons in which distributions are not elliptically symmetric. One of the new methods simply removes outliers using a projection-type multivariate outlier detection method that has been found to perform well relative to other outlier detection methods that have been proposed. The other new method belongs to the class of projection pursuit techniques and differs from other projection pursuit methods in terms of the function it tries to maximize. The comparisons include the method derived by Maronna (2005), the spherical method derived by Locantore et al. (1999), as well as a method proposed by Hubert, Rousseeuw, and Vanden Branden (2005). From the perspective used, the method by Hubert et al. (2005), the spherical method, and one of the new methods dominate the method derived by Maronna. PMID- 18411533 TI - Facial expressions of emotion (KDEF): identification under different display duration conditions. AB - Participants judged which of seven facial expressions (neutrality, happiness, anger, sadness, surprise, fear, and disgust) were displayed by a set of 280 faces corresponding to 20 female and 20 male models of the Karolinska Directed Emotional Faces database (Lundqvist, Flykt, & Ohman, 1998). Each face was presented under free-viewing conditions (to 63 participants) and also for 25, 50, 100, 250, and 500 msec (to 160 participants), to examine identification thresholds. Measures of identification accuracy, types of errors, and reaction times were obtained for each expression. In general, happy faces were identified more accurately, earlier, and faster than other faces, whereas judgments of fearful faces were the least accurate, the latest, and the slowest. Norms for each face and expression regarding level of identification accuracy, errors, and reaction times may be downloaded from www.psychonomic.org/archive/. PMID- 18411534 TI - Confounds in pictorial sets: the role of complexity and familiarity in basic level picture processing. AB - Complexity is conventionally defined as the level of detail or intricacy contained within a picture. The study of complexity has received relatively little attention-in part, because of the absence of an acceptable metric. Traditionally, normative ratings of complexity have been based on human judgments. However, this study demonstrates that published norms for visual complexity are biased. Familiarity and learning influence the subjective complexity scores for nonsense shapes, with a significant training x familiarity interaction [F(1,52) = 17.53, p < .05]. Several image-processing techniques were explored as alternative measures of picture and image complexity. A perimeter detection measure correlates strongly with human judgments of the complexity of line drawings of real-world objects and nonsense shapes and captures some of the processes important in judgments of subjective complexity, while removing the bias due to familiarity effects. PMID- 18411535 TI - Age of face matters: age-group differences in ratings of young and old faces. AB - Young (n = 24) and old (n = 24) participants rated 160 faces of young and old individuals taken from the CAL/PAL Face Database (Minear & Park, 2004) with regard to attractiveness, likeability, distinctiveness, goal orientation, energy, mood, and age. Ratings are reported for each face separately. Further analyses showed that the age groups differed in their ratings of young and old faces. On average, old participants evaluated the faces as more positive (i.e., more attractive, more energetic) than did young participants. In line with research on a negative aging stereotype, old faces were judged as less positive than young faces. They were, for instance, seen as less attractive, less likeable, less distinctive, less growth-oriented, and less energetic. The findings of the present study can serve as a basis for the selection of appropriate facial stimuli in age-comparative studies of face perception, face processing, or memory for faces. All face-specific data are archived at www.psychonomic.org/archive. PMID- 18411536 TI - Psycholinguistic norms and face naming times for photographs of celebrities in French. AB - A set of 105 photographs of celebrities has been standardized in French on distinctiveness, proper name agreement, face agreement, age of acquisition (AoA), and subjective frequency. Statistics on the collected variables for photographs are provided. The relationships between these variables have been analyzed. Face naming latencies have also been collected for the photographs of celebrities, and several multiple regression analyses have been carried out on naming latencies and percentages of tip-of-the-tongue (TOT) phenomena. Themain determinants of naming speed included AoA, face agreement, and name agreement. In addition, AoA, together with distinctiveness and face agreement, reliably predicted the percentages of TOTs. The norms, photographs of the celebrities, and spoken naming latencies corresponding to the celebrities are available on the Internet at norms.celebrities.googlepages.com and should be of great use to researchers interested in the processing of famous people. PMID- 18411537 TI - CVC syllables for investigating the phonetic sensitivity of Mandarin and English speakers. AB - Although many individual speech contrasts pairs have been studied within the cross-language literature, no one has created a comprehensive and systematic set of such stimuli. This article justifies and details an extensive set of contrast pairs for Mandarin Chinese and American English. The stimuli consist of 180 pairs of CVC syllables recorded in two tokens each (720 syllables total). Between each CVC pair, two of the segments are identical, whereas the third differs in that a segment drawn from a "native" phonetic category (either Mandarin, English, or both) is partnered with a segment drawn from a "foreign" phonetic category (nonnative to Mandarin, English, or both). Each contrast pair differs by a minimal phonetic amount and constitutes a meaningful contrast among the world's languages (as cataloged in the UCLA Phonological Segment Inventory Database of 451 languages). The entire collection of phonetic differences envelops Mandarin and English phonetic spaces and generates a range of phonetic discriminability. Contrastive segments are balanced through all possible syllable positions, with noncontrastive segments being filled in with other "foreign" segments. Although intended to measure phonetic perceptual sensitivity among adult speakers of the two languages, these stimuli are offered here to all for similar or for altogether unrelated investigations. PMID- 18411538 TI - Corpora of Vietnamese texts: lexical effects of intended audience and publication place. AB - This article has two primary aims. The first is to introduce a new Vietnamese text-based corpus. The Corpora of Vietnamese Texts (CVT; Tang, 2006a) consists of approximately 1 million words drawn from newspapers and children's literature, and is available online at www.vnspeechtherapy.com/vi/CVT. The second aim is to investigate potential differences in lexical frequency and distributional characteristics in the CVT on the basis of place of publication (Vietnam or Western countries) and intended audience: adult-directed texts (newspapers) or child-directed texts (children's literature). We found clear differences between adult- and child-directed texts, particularly in the distributional frequencies of pronouns or kinship terms, which were more frequent in children's literature. Within child- and adult-directed texts, lexical characteristics did not differ on the basis of place of publication. Implications of these findings for future research are discussed. PMID- 18411539 TI - AGL StimSelect: software for automated selection of stimuli for artificial grammar learning. AB - Artificial grammar learning (AGL) is an experimental paradigm that has been used extensively incognitive research for many years to study implicit learning, associative learning, and generalization on the basis of either similarity or rules. Without computer assistance, it is virtually impossible to generate appropriate grammatical training stimuli along with grammatical or nongrammatical test stimuli that control relevant psychological variables. We present the first flexible, fully automated software for selecting AGL stimuli. The software allows users to specify a grammar of interest, and to manipulate characteristics of training and test sequences, and their relationship to each other. The user therefore has direct control over stimulus features that may influence learning and generalization in AGL tasks. The software, AGL StimSelect, enables researchers to develop AGL designs that would not be feasible without automatic stimulus selection. It is implemented in MATLAB. PMID- 18411540 TI - Portuguese category norms for children. AB - This study presents Portuguese category norms for children of three different age groups: preschoolers (3- to 4-year-olds), second graders (7- to 8-year-olds), and preadolescents (11- to 12-year-olds). Three hundred Portuguese children (100 in each group) completed an exemplar-generation task. Preschoolers generated exemplars for 13 categories, second graders generated exemplars for 17 categories, and preadolescents generated exemplars for 21 categories. For each group, responses within each category were organized according to frequency of production in order to derive exemplar-production norms for sets of tested categories. The results also included information about the number of responses and exemplars, idiosyncratic and inappropriate responses, and commonality and diversity indexes for all the categories. A comparison of these children's norms with the Portuguese adult norms was also presented. The full set of norms may be downloaded from www.psychonomic.org/archive. PMID- 18411541 TI - Semantic feature production norms for a large set of objects and events. AB - Semantic features produced by speakers of a language when given a word corresponding to a concept have provided insight into numerous behavioral phenomena concerning semantic representation in language-impaired and -unimpaired speakers. A number of theories concerning the organization of semantic memory have used features as their starting point. Here, we provide a set of feature norms collected from approximately 280 participants for a total of 456 words (169 nouns referring to objects, 71 nouns referring to events, and 216 verbs referring to events). Whereas a number of feature norms for object concepts already exist, we provide the first set of norms for event concepts. We have used these norms (for both objects and events) in research addressing questions concerning the similarities and differences between the semantic representation of objects and events and in research concerning the interface between semantics and syntax, given that events can be expressed in language as nouns or verbs. Some of this research is summarized here. These norms may be downloaded from www.psychonomic.org/archive. PMID- 18411542 TI - ONESC: a database of orthographic neighbors for Spanish read by children. AB - In this article, we present a database of orthographic neighbors for words that Spanish children read during elementary education. The reference dictionary for lexical entries and frequencies (which had its origin in Martinez & Garcia, 2004) comprises approximately 100,000 words and is the result of accumulating the words read by a sample of children from first to sixth grades. Using the criterion for orthographic neighbors described by Coltheart, Davelaar, Jonasson, and Besner (1977), we present basic statistics related to neighborhood size as a function of the positions of divergent letters, the cumulative frequency of the neighbors, and the numbers of neighbors of higher, lower, and equal frequency. We also attempt to illustrate and unravel the nature of the relationships among the variables neighborhood size, length, and frequency in the distribution of neighbors. The database described in this article is available at www.psychonomic.org/archive. PMID- 18411543 TI - Word associations: norms for 1,424 Dutch words in a continuous task. AB - This study describes the collection of a large set of word association norms. In a continuous word association task, norms for 1,424 Dutch words were gathered. For each cue, three association responses were obtained per participant. In total, an average of 268 responses were collected for each cue. We investigated the relationship with similar procedures, such as discrete association tasks and exemplar generation tasks. The results show that the use of a continuous task allows the study of weaker associations in comparison with a discrete task. The effects of the continuous tasks were investigated for set size and the availability characteristics of the responses, measured through word frequency, age of acquisition, and imageability. Finally, we compared our findings to those of a semantically constrained version of the association task in which participants generated responses within the domain of a semantic category. Results of this comparison are discussed. The Appendix cited in this article is available at www.psychonomic.org/archive. PMID- 18411544 TI - Au pairs are rarely male: norms on the gender perception of role names across English, French, and German. AB - A list of role names for future use in research on gender stereotyping was created and evaluated. In two studies, 126 role names were rated with reference to their gender stereotypicality by English-, French-, and German-speaking students of universities in Switzerland (French and German) and in the U.K. (English). Role names were either presented in specific feminine and masculine forms (Study 1) or in the masculine form (generic masculine) only (Study 2). The rankings of the stereotypicality ratings were highly reliable across languages and questionnaire versions, but the overall mean of the ratings was less strongly male if participants were also presented with the female versions of the role names and if the latter were presented on the left side of the questionnaires. PMID- 18411545 TI - Word associations: network and semantic properties. AB - A number of properties of word associations, generated in a continuous task, were investigated. First, we investigated the correspondence of word class in association cues and responses. Nouns were the modal word class response, regardless of the word class of the cue, indicating a dominant paradigmatic response style. Next, the word association data were used to build an associative network to investigate the centrality of nodes. The study of node centrality showed that central nodes in the network tended to be highly frequent and acquired early. Small-world properties of the association network were investigated and compared with a large English association network (Steyvers & Tenenbaum, 2005). Networks based on a multiple association procedure showed small world properties despitebeing denser than networks based on a discrete task. Finally, a semantic taxonomy was used to investigate the composition of semantic types in association responses. The majority of responses were thematically related situation responses and entity responses referring to parts, shape, or color. Since the association task required multiple responses per cue, the interaction between generation position and semantic role could be investigated and discussed in the framework of recent theories of natural concept representations (Barsalou, Santos, Simmons, & Wilson, in press). PMID- 18411546 TI - A database of associative strengths from the strength-sampling model: a theory based supplement to the Nelson, McEvoy, and Schreiber word association norms. AB - The strength-sampling model of free association (Nelson, McEvoy, & Dennis, 2000) claims that the probability of word association in free-association norms results from a sampling process. For a given cue word, each response word has an underlying distribution of strength values. In the free-association task, presentation of the cue word activates a random sample of strengths, one for each response. The highest strength wins, and its response is reported. In the present work, gradient descent was used to compute the theoretical mean strengths for each cue-response pair in the Nelson, McEvoy, and Schreiber (2004) norms. The resulting database may be downloaded from www.psychonomic.org/archive/. PMID- 18411548 TI - Comparison of automated procedures for ARMA model identification. AB - This article evaluates the performance of three automated proceduresfor ARMA modelidentification commonly available in current versions of SAS for Windows: MINIC, SCAN, and ESACF. Monte Carlo experiments with different model structures, parameter values, and sample sizes were used to compare the methods. On average, the procedures either correctly identified the simulated structures or selected parsimonious nearly equivalent mathematical representations in at least 60% of the trials conducted. For autoregressive models, MINIC achieved the best results. SCAN was superior to the other two procedures for mixed structures. For moving average processes, ESACF obtained the most correct selections. For all three methods, model identification was less accurate for low dependency than for medium or high dependency processes. The effect of sample size was more pronounced for MINIC than for SCAN and ESACE SCAN and ESACF tended to select higher-order mixed structures in larger samples. These findings are confined to stationary nonseasonal time series. PMID- 18411547 TI - ML-DEs: a program for designing efficient multilevel studies. AB - The multilevel model is increasingly used as a flexible tool in the statistical analysis of dependent behavioral research data. A drawback of this model's flexibility is that it complicates designing the study. For example, an important additional consideration in the design of a multilevel study is choosing the number and the size of the clusters to sample to ensure sufficient efficiency as quantified by precision, bias, or statistical power. To help researchers in designing their multilevel study, a user-friendly simulation tool is introduced (Multilevel Design Efficiency Using Simulation, or ML-DEs), also allowing for design questions that have not been dealt with analytically in the literature, while avoiding complex specifications of simulation studies. ML-DEs generates MLwiN macros for running the simulations and handles its output using R scripts to compare the designs' efficiencies for both fixed and random parameters, allowing for small sample sizes, unbalanced data, and more than two levels. PMID- 18411549 TI - Rebus puzzles as insight problems. AB - Research on insight-the phenomenon of suddenly solving an apparently intransigent problem--has been hampered because stimulus problems have been few, ad hoc, heterogeneous, and difficult to solve. Responding to the need for a larger pool of problems of a similar type and of varying level of difficulty, we report an experiment testing the validity of rebuses as insight problems. A rebus combines verbal and visual clues to a common phrase, such as PAINS ("growing pains"). Solving a rebus requires breaking implicit assumptions of normal reading, similar to the restructuring required in insight. We hypothesized that, the more implicit assumptions are involved, the more difficult the solution. The results of a two part experiment supported the hypothesis, with participants solving more problems involving one assumption than they did problems involving two or more. Also, rebus performance correlated significantly with self-rated insight and with scores on remote associates, but not with general verbal ability. The findings suggest that rebus puzzles may be a useful source of theoretically grounded insight problems. PMID- 18411550 TI - Deciphering the secret code: a new methodology for the cross-cultural analysis of nonverbal behavior. AB - This article addresses methodological issues in the description and evaluation of nonverbal behavior (NVB) in cross-cultural studies. Videos containing cultural samples of NVB are of little use when analyzing the effects of NVB across cultures. Since video recordings confuse visible behavior with the physical appearances of the actors, they are likely to activate stereotypes derived from hints about an actor's culture and ethnicity that override the effects of the observed NVB. As a solution for this problem, we suggest here a unified computerized method for the transcription and experimental simulation of NVB. This approach makes use of advanced 3-D animation tools to generate detailed protocols of NVB that can be used to generate culture-free stimulus materials using standardized virtual characters. PMID- 18411551 TI - New and updated tests of print exposure and reading abilities in college students. AB - The relationship between print exposure and measures of reading skill was examined in college students (N = 99, 58 female; mean age = 20.3 years). Print exposure was measured with several new self-reports of reading and writing habits, as well as updated versions of the Author Recognition Test and the Magazine Recognition Test (Stanovich & West, 1989). Participants completed a sentence comprehension task with syntactically complex sentences, and reading times and comprehension accuracy were measured. An additional measure of reading skill was provided by participants' scores on the verbal portions of the ACT, a standardized achievement test. Higher levels of print exposure were associated with higher sentence processing abilities and superior verbal ACT performance. The relative merits of different print exposure assessments are discussed. PMID- 18411552 TI - Developing a generic approach to online automated analysis of writing and drawing tests in clinical patient profiling. AB - Writing and drawing tests are widely used in the clinical environment for the diagnosis of a variety of neuropsychological conditions. Conventional assessment of these tests involves the inspection by trained assessors of the completed patient response. This article describes the development of a computer-based framework for data capture, automated feature analysis, and result reporting for a range of drawing- and writing-based test batteries. In developing this framework, we have exploited the commonality between tasks while allowing for both flexibility in configuration across condition-specific testing requirements and extensibility for future test development. Using the two example clinical conditions of visuospatial neglect and dyspraxia, we illustrate the advantages of utilizing a computer-based analysis system, describe a structured approach to system implementation, and demonstrate the generality of this implementation for different conditions of interest, which extends to feature selection and design. PMID- 18411553 TI - Creating physically accurate visual stimuli for free: spectral rendering with RADIANCE. AB - Visual psychophysicists, who study object, color, and light perception, have a demand for software that produces complex but, at the same time, physically accurate stimuli for their experiments. The number of computer graphic packages that simulate the physical interaction of light and surfaces is limited, and mostly they require the purchase of a license. RADIANCE (Ward, 1994), however, is freely available and popular in the visual perception community, making it a prime candidate. We have shown previously that RADIANCE's simulation accuracy is greatly improved when color is coded by spectra, rather than by the originally envisaged RGB triplets (Ruppertsberg & Bloj, 2006). Here, we present a method for spectral rendering with RADIANCE to generate hyperspectral images that can be converted to XYZ images (CIE 1931 system) and then to machine-dependent RGB images. Generating XYZ stimuli has the added advantage of making stimulus images independent of display devices and, thereby, facilitating the process of reproducing results across different labs. Materials associated with this article may be downloaded from www.psychonomic.org. PMID- 18411554 TI - Prevalence and reporting of near-point binocular convergence and acuity vision deficits in potential research participants. AB - Visual abilities at near-point distances of 200 self-reported normal vision college students were assessed. Results show that normal visual functioning with no deficits is the exception. The most common visual deficits observed involved binocular lateral posture and fusion convergence deficits. Binocular and monocular acuity deficits also had high prevalence rates, although not as prevalent as the binocular convergence deficits. In addition, 1,340 articles published in four major journals from the years 1997-2004 were examined to ascertain the extent to which authors report the visual capabilities of their participants. Reporting of research participant visual abilities by authors using visual stimuli in experiments appears to be lacking. The results are discussed in terms of the potential impact that visual deficits may have on results obtained in experiments in which visual stimuli are used and the importance of reporting the visual assessment tests and procedures used to assess the visual abilities of potential participants. PMID- 18411555 TI - Affective auditory stimuli: characterization of the International Affective Digitized Sounds (IADS) by discrete emotional categories. AB - Although there are many well-characterized affective visual stimuli sets available to researchers, there are few auditory sets available. Those auditory sets that are available have been characterized primarily according to one of two major theories of affect: dimensional or categorical. Current trends have attempted to utilize both theories to more fully understand emotional processing. As such, stimuli that have been thoroughly characterized according to both of these approaches are exceptionally useful. In an effort to provide researchers with such a stimuli set, we collected descriptive data on the International Affective Digitized Sounds (IADS), identifying which discrete categorical emotions are elicited by each sound. The IADS is a database of 111 sounds characterized along the affective dimensions of valence, arousal, and dominance. Our data complement these characterizations of the IADS, allowing researchers to control for or manipulate stimulus properties in accordance with both theories of affect, providing an avenue for further integration of these perspectives. Related materials may be downloaded from the Psychonomic Society Web archive at www.psychonomic.org/archive. PMID- 18411556 TI - The high-hurdle technique put to the test: failure to find evidence that increasing loading times enhances data quality in Web-based studies. AB - Two Web-based experiments examined the usefulness of artificially delaying the loading of the first page of the study. The idea pursued in this technique is to filter out less-motivated respondents through a higher respondent burden in the form of waiting time. Participants who remain in the study despite having had to wait for the first page of the study to appear on the screenare expected to be more highly motivated, and thus to produce data of higher quality. In both experiments, as expected, the longer the loading time, the lower the likelihood of people responding to the study. However, contrary to expectation, the dropout rate and quality of data were independent of the loading time. Therefore, artificially delaying the loading of the first page of the study is counterproductive. PMID- 18411557 TI - Telemetric monitoring in the behavior sciences. AB - This article reviews recent advances in telemetrics, a class of wireless information systems technology that can collect and transmit a wide variety of behavioral and environmental data remotely. Telemetrics include wearable computers that weave on-body sensors into articles of clothing, ubiquitous computers that embed sensors and transmitters seamlessly into the environment, and handheld devices, such as mobile phones and personal digital assistants, that can record cognitive and affective states. Examples of telemetric applications are provided to illustrate how this technology has been used in the behavioral sciences to unobtrusively and repeatedly gather physiological, behavioral, environmental, cognitive, and affective data in natural settings. Special issues relating to privacy and confidentiality, practical considerations, and statistical and measurement challenges when telemetrics are used are also discussed. PMID- 18411558 TI - Development of a magnetic-resonance-compatible photoplethysmograph amplifier for behavioral and emotional studies. AB - The purpose of the present study was to develop a magnetic resonance (MR) compatible photoplethysmograph (PPG) system that can measure the raw PPG signal during MR image acquisition. The system consists of an optic sensor that measures the optic signal, an optic cable that transmits a near-infrared optic signal, a signal amplifier, and a filter for noise removal. To minimize interactive noise, only the optic cable and the optic sensor module are located inside the MR room; the signal amplifier and filter are located outside the MR room. An experiment verified that a reliable PPG signal can be obtained without causing a deterioration in the MR image. PMID- 18411559 TI - The sensitivity of a palm-based psychomotor vigilance task to severe sleep loss. AB - In this study, we evaluated the sensitivity of a 5-min personal digital assistant psychomotor vigilance test (PDA-PVT) to severe sleep loss. Twenty-one participants completed a 10-min PVT-192 and a 5-min PDA-PVT at two hourly intervals during 62 h of sustained wakefulness. For both tasks, response speed and number of lapses (RTs > 500) per minute significantly increased with increasing hours of wakefulness. Overall, standardized response speed scores on the 5-min PDA-PVT closely tracked those of the PVT-192; however, the PDA-PVT was generally associated with more lapses/minute. Closer inspection of the data indicated that when the level of sleep loss and fatigue became more severe (i.e., Day 3), the 5-min PDA-PVT was not quite as sensitive as the 10-min PVT-192 when 2 to 10-sec foreperiods were used for both. It is likely, however, that the observed differences between the two devices was due to differences in task length. Thus, the findings provide further evidence of the validity of the 5-min PDA-PVT as a substitute for the 10-min PVT-192, particularly in circumstances in which a shorter test is required and/or the PVT-192 is not as practical. PMID- 18411560 TI - Maze Suite 1.0: a complete set of tools to prepare, present, and analyze navigational and spatial cognitive neuroscience experiments. AB - Maze Suite is a complete set of tools that enables researchers to perform spatial and navigational behavioral experiments within interactive, easy-to-create, and extendable (e.g., multiple rooms) 3-D virtual environments. Maze Suite can be used to design and edit adapted 3-D environments, as well as to track subjects' behavioral performance. Maze Suite consists of three main applications: an editing program for constructing maze environments (MazeMaker), a visualization/rendering module (MazeWalker), and an analysis and mapping tool (MazeViewer). Each of these tools is run and used from a graphical user interface, thus making editing, execution, and analysis user friendly. MazeMaker is a .NET architecture application that can easily be used to create new 3-D environments and to edit objects (e.g., geometric shapes, pictures, landscapes, etc.) or add them to the environment effortlessly. In addition, Maze Suite has the capability of sending signal-out pulses to physiological recording devices, using standard computer ports. Maze Suite, with all three applications, is a unique and complete toolset for researchers who want to easily and rapidly deploy interactive 3-D environments. PMID- 18411561 TI - Diagnostic test evaluation of a nasal flow monitor for obstructive sleep apnea detection in sleep apnea research. AB - In this diagnostic test evaluation of a nasal flow monitoring device for obstructive sleep apnea (OSA), 34 patients referred for polysomnography were studied at home for three consecutive nights with the monitor. The mean age of subjects (+/-SD) was 41.9+/-10.3 years, and their mean apnea-hypopnea index (AHI) was 31.5+/-27.2. The difference between the average AHI from three nights at home on the monitor and the polysomnogram (PSG) result was 1.8+/-17.1. The area under the receiver operating characteristic curve (AUC) for PSG AHI > or = 10 was .96. With a threshold AHI of 18 on the flow monitor, sensitivity was .92, specificity .86, positive predictive value .96, and negative predictive value .75. For detecting severe OSA (AHI > or = 30), the AUC was .85. With knowledge of appropriate thresholds and the pretest risk of OSA, the flow monitor can be used to detect or exclude OSA for sleep-related research, as well as to identify severe cases needing priority for further evaluation. PMID- 18411562 TI - An n-back task using vibrotactile stimulation with comparison to an auditory analogue. AB - We report a vibrotactile version of the common n-back task used to study working memory. Subjects wore vibrotactile stimulators on three fingers of one hand, and they responded by pressing a button with the other hand whenever the current finger matched the one stimulated n items back. Experiment 1 showed a steep decline in performance as n increased from 1 to 3; each additional level ofn decreased performance by 1.5 d' units on average. Experiment 2 supported a central capacity locus for the vibrotactile task by showing that it correlated strongly with an auditory analogue; both tasks were also related to standard digit span. The vibrotactile version of n-back may be particularly useful in dual task contexts. It allows the assessment of cognitive capacity in sensory-impaired populations in which touch remains intact, and it may find use in brain-imaging studies in which vibrotactile stimuli impose a memory load. PMID- 18411563 TI - Medical malpractice and court allegations in Switzerland. PMID- 18411564 TI - Health care cost containment. PMID- 18411565 TI - The resistible growth of health care costs. AB - Rather than our routinely blamed ageing demography, pharmaceutical promotion and the medical business, not research, are responsible for our ever growing health bill. To keep essential health care affordable, only what has been proved necessary and cost effective should be financed by some kind of risk mutualisation system. Hedonistic care should be left to the free market. From conception to death, a devastating culture of medicalization and therapeutic agressivity has turned naturally inexpensive processes, such as conception, birth, ageing and death, into over-priced medical achievements. The increasing lack of personal and social responsibility triggered by the market, such as junk food, tobacco, drugs, sedentarity or trash media, multiply life-threatening illnesses such as diabetes 2, obesity, cardiovascular diseases and all kinds of cancers. Screenings require millions of participants and intense statistical analysis to prove any efficacy. Screenings, testings and proactive practices make people sick and produce more patients than they save lives , while generating exceptional returns on investments thanks to state and insurance financing; they should be put under public control. New drugs are unaffordable in spite of their dubious efficacy which often relies on biased and underpowered studies. Because they target desperate, debilitating, up to now incurable diseases like metastatic cancers, multiple sclerosis, Alzheimer, polyarthritis, Crohn disease, patients and their families want them by any means and at any price. The answer to the North-South health gap is in a global deal: a declining demographic trend, already well under way and free circulation not only of goods but also of people which would in the long run shape up the age pyramid of a progressively mixed population. That could also save lives at both ends of the human chain: those who die from starvation and those who die from overfeeding. PMID- 18411566 TI - Abdominal Transplant Surgery and Transplant Coordination University Hospitals Leuven 1997-2007: an overview. AB - The transplant surgery and transplant coordination department was created in 1997 to meet up with the demand of the growing abdominal transplant surgery and organ procurement activity at the University Hospitals in Leuven. Since then, the procurement activity has increased and is currently distributed within the University Hospital Gasthuisberg and a network of approximately 25 collaborative hospitals. The profile of the donors has changed with older donors and more co morbidity factors (obesity, hypertension, etc.). This donor activity represents approximately 30% of the national donor pool. Over the last 10 years, more than 1100 kidneys, more than 500 livers, approximately 50 pancreas, and 5 intestines have been transplanted in both adults and children. One year survival equal to- or exceeding 90% has been achieved for all abdominal organs and this compares favorably with international registries. More than 40 multi-visceral transplants {liver in combination with abdominal (kidney, pancreas, intestine) or thoracic (heart, double lung, heart-lung) organs} have been performed with results equivalent to isolated liver transplants and very little immunological graft loss (probably due to the immunoprotective effect of the liver). A live donation program was started for the kidney (40 cases) and for the liver (10 cases) in adults and children and no surgical graft loss has been seen so far. Introduction of new machine perfusion systems (and development of donor protocols) has made it possible to restart a non-heart-beating donor program for kidney transplantation. Experimental demonstration that livers tolerate short periods of warm ischemia has also allowed to start liver transplantation from non-heart-beating donors. In the future, machine perfusion of livers, viability testing, and biological modulation are likely to widen the use of marginal livers for transplantation and improve the results. An immunomodulatory protocol proven in the lab to induce the development of regulatory T cells has been applied clinically to 5 consecutive intestinal transplants. All 5--at the time of writing--have been rejection-free and have achieved nutritional independence. Continuous research and development is warranted to increase the organ donor pool (currently the solely limiting factor of transplantation) and to optimize long-term graft and patient outcome. PMID- 18411567 TI - Importance of informatics and database management in transplant coordination. AB - Advanced use of informatics within modern health care has become essential. Transplantation and transplant coordination, a high technically advanced and very specific niche within health care, is strongly depending on time management and exclusion of possible pitfalls within an acute organization at both the donor and the recipient sites. Based on the increased donor and transplant activity, we stratified two goals. The first goal was to improve working methods at the donor site. The second goal was to reduce administrative tasks and increase quality follow-up at the recipient side. For the donor process, we designed a Donor Database, that was created for donor registration and quality data reporting. A 24/24 h accessible website was created and was linked with clinical pathways and reports. For the liver transplant process, we built another database system in FileMaker pro, creating a quality follow-up and reporting methods. Based on a retrospective analysis and review of two executive time periods, we saw a clear improvement in the donor reporting method, and the quality of the procedure. Possible mistakes within the acute organization were easily detected based on clinical pathways provided by the website on one hand, and integrated within the database system on the other hand. We succeeded in bringing high-quality informatics to the floor of donor and transplant procedures and follow-up. Retrospective analysis showed a definite improvement, with a positive impact on data reporting, time management and administrative follow-up. PMID- 18411568 TI - What is the limiting factor for organ procurement in Belgium: donation or detection ? What can be done to improve organ procurement rates? AB - In trying to overcome the growing gap between demand and offer of organs for transplantation, solutions are usually searched for by comparing successful and unsuccessful models in different countries. In particular, one element in the more successful countries such as for instance presumed consent, or one element in the less successful countries such as for instance refusal by relatives, are seen as possible reasons for these differences. This article tackles the problem of organ donor shortage through a new multi-level approach. Organ donation can indeed be analyzed on three different levels: the macro-level, the meso-level and the micro-level. The macro-level refers to the governmental structure where legislation, policies and funding are three essential elements necessary to make donation possible. The meso-level refers to the health care organization and the professionals who surround the process of organ donation and transplantation. Facilitating this process through standardized protocols and improving detection of organ donors are the two major elements. The micro-level refers to the individual believes and personal attitudes towards organ donation. This new multi level approach gives a thorough and complete analysis of problems and allows to propose potential solutions to try to overcome the chronic organ shortage. PMID- 18411570 TI - Donor categories: heart-beating, non-heart-beating and living donors; evolution within the last 10 years in UZ Leuven and Collaborative Donor Hospitals. AB - Over the past 10 years, the University Hospitals Leuven and their group of Collaborative Donor Hospitals (approximately 20) have tried to maximize their contribution to the national and Eurotransplant donor pool. In this time period, 1042 potential donors and 703 effective donors were coordinated and their organs allocated through Eurotransplant. This activity represented approximately 30% of the national donor pool and approximately 32% of the national organ pool. For Belgium, the non-heart-beating donor activity represented 11.38% of all donors in 2006. Since 1997, 167 potential live donors have been screened in our center. Of these, 48 transplants (28.74%) (39 kidneys--9 livers) have been performed. A boost of screened candidates was seen over the last 3 years, with a 500% increase of records being evaluated. Although the Belgian live donation activity remains one of the lowest in the world, there has been a clear increase over the last 3 years with about 10% of all kidney transplant activity originating now from live donors. PMID- 18411569 TI - Evolution in allocation rules for renal, hepatic, pancreatic and intestinal grafts. AB - Organ transplantation is the victim of his own success. The results of transplantation are excellent and more patients are activated on the waiting list. The need for organs exceeds the supply. Which criteria are used to allocate available grafts to patients on the waiting list ? Organ allocation and finding the "best match" between donor and recipients, is the goal of Eurotransplant, the organ sharing organization for seven European countries (Austria, Croatia, Germany, Luxemburg, Slovenia, The Netherlands and Belgium). Last decade, the allocation system has switched from a "center-driven" (organ allocated to a center) to a "patient-driven" system (organ allocated to a particular patient). For the allocation of abdominal organs some general allocation rules are followed: blood group compatibility, priority for high urgencies. The allocation of kidneys is based on a point score system based on waiting time, HLA and donor location (to reduce the cold ischemia time). In addition to this standard allocation procedure, there are still specific procedures for pediatric recipients and for candidates > or = 65 year old. There is also an "acceptable" mismatch program for recipients at high immunological risk. The liver allocation system recently changed and is now based on the MELD score, a formula that calculates the probability of death within 3 months on the waiting list. For pancreas and intestine, the system is based on blood group, medical urgency, waiting time, donor region and weight (for intestine). PMID- 18411571 TI - Living donor kidney transplantation: analysis of the first 40 cases performed in UZ Leuven. AB - Living donation kidney transplantation has been popular worldwide to try to increase the donor pool. In Belgium, the rate of living donation kidney transplantation has been traditionally relatively low compared to other countries. This is--in part--due to the relatively higher cadaveric organ offer that is available in Belgium (around 25 donors per million inhabitants), compared to other countries. However, the increasing waiting times on cadaveric waiting list and the superiority of the results of live donation versus cadaveric kidney transplantation have led to a reappraisal of this strategy. In our center a living donation kidney transplant programme was started in 1997. Since then 40 cases of live donation kidney transplantation have been performed and are reported herein. PMID- 18411572 TI - Liver transplantation from non-heart-beating donors: current status and future prospects in an experimental model. AB - The widening gap between supply and demand for liver transplantation has prompted many transplant centers to use donors after cardiac death or non-heart-beating donors. These livers--contrary to "classical" brain-dead donors--are exposed to an unavoidable period of warm ischemia, jeopardizing graft function post transplantation. In a newly developed preclinical model of liver transplantation, we studied--in a biologically unmodified environment--the exact tolerance of the liver to warm ischemia. Following the evidence that liver transplantation from non-heart-beating donors is feasible and safe, provided that warm and cold ischemia are kept short, a clinical programme of liver transplantation from non heart-beating donors was successfully initiated in our and other Belgian centers. Recently, we demonstrated that the tolerance of such livers to warm ischemia could be substantially improved when some of the previously identified mechanisms leading to graft non-function were tackled by a multi-factorial pharmacological strategy. Meanwhile, cold storage has proven to be insufficient to optimally preserve organs from non-heart-beating donors. As an alternative, machine perfusion preservation was found to consistently improve outcome in kidney transplantation from non-heart-beating donors. Similarly, machine perfusion preservation could improve the preservation of livers, allowing to predict viability prior to transplantation and to ameliorate tolerance to warm ischemia. At present, the definition and development of optimal machine perfusion settings are under investigation at our institution. PMID- 18411573 TI - Intestinal transplantation: from the laboratory to the clinics. AB - The intestine has long been seen as a "forbidden" organ to transplant. This is because the first attempts at Intestinal Transplantation (ITx) were defeated by rejection, technical problems, infection and graft versus host disease. Results of ITx have improved in the short-term (70 to 80% 1-year patient survival) but remain inferior to other solid organ transplants in the long-term (5 years patient survival of 50% or less). Reasons for this difference between intestine and other organ transplants are reviewed. Development of immunomodulatory protocols--e.g. protocols aiming at reducing the rejection response and facilitating engraftment--are described. Our center experience with a consecutive series of five intestinal transplants utilizing a new protolerogenic protocol and low immunosuppression is described. At time of writing, these five patients are rejection-free, nutritionally independent and lead a normal life. PMID- 18411574 TI - Pancreas transplantation: towards minimization of technical graft loss. AB - The objective of the study is to determine whether evolution in surgical techniques results in minimization of technical graft loss after simultaneous pancreas-kidney transplantation (SPKTx). Eighty consecutive patients with insulin dependent diabetes mellitus and end-stage nephropathy, and who received SPKTx, were studied. Patient, pancreas and kidney survival at 5 years were 89, 84 and 84% respectively. The porto-systemic venous anastomosis laterally to the arterial vessels resulted in exclusion of technical graft loss due to venous thrombosis. The switch from exocrine bladder drainage to enteric drainage lowered significantly the posttransplant surgical morbidity. This series indicates that excellent short- and long-term survival can be obtained after SPKTx and this with a low perioperative surgical morbidity. PMID- 18411575 TI - Milestones in pancreas transplantation in Belgium. PMID- 18411576 TI - The history of the EuroSPK - Study Group. AB - The EuroSPK Study group was created during the 4th Spitzingsee 1997 workshop in Kuhtai, Austria. Thanks to W. Land for the incentive to gather European Centres- with Switzerland and Israel--and propose them to joint efforts and share data in the field of pancreas transplantation. Today, two prospective randomized studies have been already performed; a lot of data and results have been generated and worldwide spread. The spirit of the group will continue with a new interest in innate immunity and prevention of the ischemic reperfusion injury in pancreas transplantation. PMID- 18411577 TI - Beta-cell replacement by transplantation in diabetes mellitus: when pancreas, when islets; how to allocate deceased donor pancreases in 2007? PMID- 18411578 TI - Emerging science, emerging ethical issues: who should fund innate alloimmunity suppressing drugs? AB - An emerging body of evidence suggests that the innate immune system plays a critical role in allograft rejection. Any injury to the donor organ, e.g. the reperfusion injury, induces an inflammatory milieu in the allograft which appears to be the initial event for activation of the innate immune system. Injury induced intragraft damage- associated molecular patterns (DAMPs) are recognized by donor-derived and recipient-derived, TLR4/2-bearing immature dendritic cells (iDCs). After recognition, these cells mature and initiate allorecognition/alloactivation in the lymphoid system of the recipient. Indeed, the key "innate" event, leading to activation of the adaptive alloimmune response, is the injury-induced, TLR4-triggered, and NFkappaB-mediated maturation of DCs ("innate alloimmunity"). Time-restricted treatment of innate immune events would include 1) treatment of the donor during organ removal, 2) in-situ/ex-vivo treatment of the donor organs alone, and 3) treatment of the recipient during allograft reperfusion and immediately postoperatively. Treatment modalities would include 1) minimization of the oxidative allograft injury with the use of antioxidants; 2) prevention of the TLR4-triggered maturation of DCs with the use of TLR4-antagonists; 3) inhibition of complement activation with the use of complement inhibiting agents. According to data from clinical and experimental studies it can be assumed that successful suppression of innate alloimmune events results in either subsequent significant reduction in, or even complete avoidance of the currently applied adaptive alloimmunity-suppressing drugs. However, in view of the time-restricted period of treatment, and the fear to potentially destroy its own business with currently applied alloimmunity-suppressing drugs, the pharmaceutical industry is still, but quite legitimately, reluctant to invest in the high cost of clinical development of those drugs for transplant patients because there are no marketing interests. On the other hand, clinical development of innate alloimmunity-suppressing drugs is urgently warranted. But: Who should fund? In this article, three options are explored which may contribute to a solution of the problem: 1) provision of incentives to companies for drug development; 2) conduction of clinical trials in developing countries; and 3) creation of a public-private professional partnership in analogy to the "European Rare Diseases Therapeutic Initiative" (ERDITI). We suggest and recommend the creation of such a partnership which may be called: "The European Initiative for the Suppression of Innate Alloimmunity" ("EISIA"). In analogy to ERDITI, the main goals of this organization should be:--to provide a streamlined facilitated process of collaboration between Academic Teams/Transplant Centres, Study Groups, and Pharma Companies to develop innate alloimmunity-suppressing drugs;--to give Academic Teams/Transplant Centres facilitated access to a large variety of compounds, developed by companies for other indications, which can be evaluated pre-clinically and, if warranted, clinically;--to guarantee the continuity all the way from research to development and commercialisation of the drug. If preclinical studies uncover the potential of a compound for suppressing innate alloimmune events, the Pharma Partner who has rights to this compound will either develop himself the drug for organ transplantation indication or allow its development by the academic team or a third party if he has no intentions of developing himself. PMID- 18411579 TI - Desire for a child and breast cancer. AB - About 10% of breast cancers occur in women who are younger than 40 years of age. For many of them, the breast cancer diagnosis will occur when they are still planning pregnancy. Most breast cancers are diagnosed at an early stage of the disease, i.e. stage I or II, which is associated with a high survival rate (5 years-survival ranging between 97% and 79% respectively) (1). Many of these patients will use adjuvant endocrine therapy. This treatment has no direct impact on their fertility, but postpones a possible pregnancy, since pregnancy is contra indicated during Tamoxifen treatment. On the other hand, chemotherapy increases the risk of premature ovarian failure, of early menopause, and of definitive sterility. This may result in an increased risk of depression and impaired quality of life. Furthermore, those women who remain fertile will often be advised to avoid pregnancy in the near future, in order to ensure the absence of breast cancer recurrence. Nevertheless, fertility decreases with age. Possible strategies, which permit optimal treatment of breast cancer and maintain the possibility of pregnancy, should be systematically discussed with the patient as soon as possible during treatment planning (2). Gynecologists and surgeons should encourage such patients to participate in multi-center studies evaluating strategies to preserve their fertility. Life continues after cancer; the prospect of pregnancy and child birth are part of a positive project. PMID- 18411580 TI - Yield of routine imaging after curative colorectal cancer treatment. AB - BACKGROUND/AIM: The use of imaging in the follow-up of patients after curative colorectal cancer resection is much debated. The American Society of Colon and Rectal Surgeons did not recommend routine imaging. This retrospective study assesses the yield of routine imaging to detect recurrent disease. METHODS: In 1998, 108 consecutive patients underwent curative resection for colorectal carcinoma. Minimum followup in our institution was 3 years. Multidisciplinary follow-up at a joint clinic consisted out of a history, clinical examination, serum carcinoembryonic antigen (CEA), chest X-ray and abdominal ultrasound, at least every 6 months. Colonoscopy was performed within 1 year after operation and every 3 to 5 years thereafter. The incidence, timing, means of detection and resectability of recurrence were studied. RESULTS: The recurrence rate was 22% (24 patients): liver metastases (11), extra-hepatic recurrence (10) and combined recurrence (3). Recurrent disease occurred in stage II or III cancer, except for two patients. It was diagnosed at a median of 21.5 months (range 4-79) after surgery. Means of detection were: symptoms in 2 (peritoneal disease, 8%), increasing CEA in 15 (63%), routine imaging in 6 (25%), and abdominal CT-scan in one patient. Curative resection of recurrent disease was possible in ten patients (42%): in 6/15 recurrences detected by CEA, in 3/6 recurrences detected by routine imaging, in 1 liver metastasis detected by CT and in none of the symptomatic patients. CONCLUSIONS: A CEA level increasing above 5.0 microg/L was the most important diagnostic tool. However, one quarter of the recurrences were detected by routine imaging and half of them could be resected for cure. These data support routine imaging during follow-up. PMID- 18411581 TI - The effect of distance on the postoperative follow-up of patients with breast carcinoma. AB - PURPOSE: To assess the effect of in or out of city residence of patients with breast carcinoma, where breast surgery unit treatment and follow-up is made postoperatively. METHOD: 234 patients operated on for breast carcinoma at the Breast Surgery Unit were retrospectively studied. Patients were divided into two groups; patients living in the major city where the Breast Surgery Centre is located and patients living in smaller cities, districts, towns and villages out of the city. The distance of patients' residences from the Breast Centre has also been determined in kilometres. The number of patients and the frequency of check up visits were compared in both groups. RESULTS: The number of patients residing in the city centre where the Breast Unit is located was 156 (66.7%). Comparing the frequency of patients' visits for check-up during the postoperative period, there were no differences between the two groups during the first four years. However, the patients living out of the city did not visit the Breast Unit for check-ups during the fifth postoperative year. Moreover, when the patients were classified into two groups with known and unknown outcomes, it was observed that those patients with unknown outcomes lived further away from the city where the Breast Surgery Unit was located compared to those with known outcomes (p = 0.002). DISCUSSION: Living within or out of the major city centre where the Breast Surgery Unit is located does not have any effect on the frequency of follow-up visits or the number of patients applying for check ups during the first four years postoperatively. However, there were gradual decreases over the course of time in both groups and these differences became apparent during the 5th year. In addition to this, the distance was also found to be an important factor for patients with unknown outcomes in the present study. The combination of living outside the city where The Breast Unit was located and the distance may have a negative effect on follow-ups. There is a need for new, larger scale, studies with longer follow-ups to show how this difference will change over a longer time period. PMID- 18411582 TI - Fistula and stenosis after 135 (pharyngo)laryngectomies. AB - BACKGROUND: Despite progress made with reconstruction, morbidity is still elevated after (pharyngo)laryngectomy. The present study was designed to determine the incidence and predisposing factors of the complications following (pharyngo)laryngectomy. METHODS: Primary surgical treatment was delivered in 60 patients. Seventy-five patients underwent surgical salvage following radiotherapy. Different factors were evaluated as potentially predisposing to fistula formation. RESULTS: Stenosis is rare: 5.1% in the present series. A pharyngocutaneous fistula developed in 48.8% of patients. After a multivariate analysis, the site of the tumour was defined as a significant risk factor for pharyngocutaneous fistula formation. CONCLUSIONS: Despite progress made with reconstruction, morbidity is still elevated after major resection of the phary golarynx. Stenosis, a frequent complication when partial pharyngectomy is needed, is rare: 5.1% in the present series. Fistulas are relatively frequent but the majority resolved either with local irrigation or with subsequent flaps. The site of the tumour was a significant risk factor for pharyngocutaneous fistula formation, as defined by a multivariate analysis. PMID- 18411583 TI - Extended transsternal thymectomy for myasthenia gravis: a report of 19 consecutive cases. AB - BACKGROUND: Thymectomy is considered as an effective therapeutic option for patients with myasthenia gravis (MG). This study reports the experience of our centre's investigation into the efficacy and the safety of the procedure and the influence of different pre-operative factors on the surgical outcome. METHODS: A retrospective chart review/interview was made of 19 consecutive patients who underwent extended transsternal thymectomy for MG from 1992 to 2003. The severity of the disease was determined according to the Osserman Classification. Efficacy was measured by determining the change in clinical status, the rate of remission during follow-up, and the reduction in medication requirements after thymectomy. Complete remission (CR) was defined as asymptomatic off medication for 6 months. The CR rate was calculated using the Kaplan-Meyer method. RESULTS: The mean age of the patients at surgery was 34 years (range, 9-63) and 78.9% were female. Mean length of follow up was 86 months (range, 24-163). The overall complication rate was 10.6% (1 episode of atrial fibrillation and a left recurrent laryngeal nerve palsy that resolved after the first postoperative month). There was no operative mortality. The mean hospital stay was 9.4 days (range, 5-23). The crude CR rate was 32% (n = 6). The Kaplan-Meier estimate of CR was 42% at 6 years. Age, gender, duration of symptoms, thymic histology, Osserman stage and the presence of thymoma were not identified as prognostic variables. The average daily dose of Medrol and Mestinon decreased significantly between the pre-operative period and the last follow-up (Medrol, p = 0.0081; Mestinon, p = 0.0013). CONCLUSIONS: Transsternal thymectomy for MG is safe and effective. It benefits patients with MG at all stages. Patients with thymoma are not associated with poorer remission rates. Complete responses are durable, as the CR rate remains stable over time. PMID- 18411584 TI - Surgery of carotid body tumour: 14 cases in 7 years. AB - OBJECTIVE: The goal was to review our experience in the management of carotid body tumours. MATERIAL AND METHODS: A retrospective study was performed of patients in whom carotid body tumour was diagnosed between 1998 and 2005. Data were retrieved from hospital discharge files. RESULTS: Fourteen patients were operated on. There were five patients in Shamblin class I, 4 in class II, and 5 in class III. Duplex examination was performed in all patients. Computerized tomography scans were performed in eight (57%) patients and magnetic resonance imaging scans in five patients (36%). Angiography was performed in all patients, preoperative embolization was attempted in 5 (36%). The blood loss for these patients was not less than for those without embolization. Three patients (21%) had postoperative cranial nerve deficits. All the deficits resolved. The internal carotid artery was injured in two patients and the external carotid artery was injured in three patients (36%). No stroke occured. CONCLUSION: Surgical resection is the treatment of choice for carotid body tumours. Observation of these tumours is not recommended because progressive growth is associated with increased risk of neurological deficits. Early surgical management is recommended to avoid neurological deficit due to a Shamblin class III tumour. We also do not recommend embolization. PMID- 18411585 TI - Granular cell tumour of the male breast. AB - Granular cell tumour (GCT) is a rare neoplasm that can be found in multiple sites throughout the body. Occasionally GCT is located in the breast. In general, it appears as a singular benign lesion, although it can be multi-focal and rare cases with malignant behaviour have been reported. We report a rare case of granular cell tumour of the nipple in the male breast, treated by wide local excision. PMID- 18411586 TI - Lipid-rich carcinoma of the breast: a case report. AB - Lipid-rich carcinoma is a rare variant and accounts for < or = 2% of all breast cancer diagnoses. We report a case occurring in a 53-year-old female. The patient presented with a painless, right breast mass. Clinical examination and mammography suggested malignancy. Subsequent modified radical mastectomy revealed the diagnosis of lipid-rich carcinoma. The morphological features, differential diagnosis and treatment along with a brief review of the literature are discussed in this article. Lipid-rich carcinoma (L-RC) is a very rare variant of breast carcinomas with an aggressive clinical course and poor prognosis. It presents only 1% to 2% of all breast cases. It is classified as a specific variety of mammary carcinoma because the tumour cells possess abundant vacuolated cytoplasm which is strongly positive when stained for neutral fat. Aboumrad first described it in 1963 as lipid-secreting carcinoma. However, Ramos and Taylor renamed it as lipid-rich breast carcinoma. In China, the first case was reported in 1984. Herein, we report a case of lipid-rich carcinoma occurring in a 53-year-old female patient, and the literature is reviewed. PMID- 18411587 TI - Endovascular treatment of axillary artery dissection following anterior shoulder dislocation. AB - Injury to the axillary artery is a rare complication of anterior shoulder dislocation. Open surgical repair is technically demanding because of the anatomical position of the vessel and the propensity for concomitant injuries. Standard surgical exposure techniques involve extensive dissection, including a combination of supraclavicular or infraclavicular incision, median sternotomy, and thoracotomy causing significant morbidity and mortality rates. Endovascular techniques may offer an alternative to these surgically demanding procedures. We present a patient with a traumatic dissection of the axillary artery following anterior shoulder dislocation who was successfully managed with an endovascular stent. PMID- 18411588 TI - Plexus injury after reduction of anterior caudal dislocation of the shoulder. AB - The case is reported of a 45-year-old male patient who fell when his ladder slipped away underneath him. He sustained a bilateral dislocation of the shoulders (bilateral luxatio erecta humeri). The injury on the one hand a glenohumeral dislocation of the humerus head right to ventral and caudal, accompanied by a suspected fracture of the tuberculum major and on the other hand a glenohumeral dislocation of the left humerus head to ventral and caudal, accompanied by a suspected total collum fracture. Based on this case and the pertinent literature, the patho-physiology, diagnosis and treatment of this rare injury are discussed. After immediate closed reduction, soft tissue damage, fractures or neurovascular lesions should be operated on as soon as possible. PMID- 18411589 TI - Double sigmoid colon perforation by a migrated biliary stent. AB - The case of a female patient with a covered double perforation of the sigmoid colon is described. The perforation was due to a migrated plastic biliary stent inserted 3 weeks earlier for benign biliary obstruction and was facilitated by the presence of expressed diverticulosis of the colon. The lack of peritonitis and the early surgical procedure allowed a successful primary reconstruction after the colonic resection. Since the use of biliary stents is constantly increasing, such complications as stent migration and perforation should always be kept in mind by surgeons. PMID- 18411590 TI - Caecum and ascending colon herniated in the lesser sac through the Winslow's hiatus. A case report. AB - Herniation of the caecum and the ascending colon in the lesser sac through the Winslow's hiatus is an uncommon cause of intestinal occlusion. The clinical presentation can be acute or chronic. The diagnosis often remains unclear before surgical operation, even with the assistance of abdominal CT, providing an image of a digestive structure in the lesser sac and the presence of mesentery between the portal triad and the vena cava. The treatment consists of reducing the internal hernia, which could be considered under laparoscopy. PMID- 18411591 TI - Torsion of the gallbladder: two case reports. AB - Torsion of the gallbladder is a rare disease. It is simply defined as a rotation of the gallbladder on its mesentery along the axis of the cystic duct and cystic artery. The aetiology is unknown. However, several factors are postulated as playing causative roles. The disease symptoms mimic acute cholecystisis. The current value of radiological imaging is limited, but clinically, volvulus of the gallbladder is an acute surgical situation. It may be treated and diagnosed by a laparoscopic approach. PMID- 18411592 TI - Pericardiotomy: the first cardiac operation. PMID- 18411593 TI - Spinal cord stimulation. New regulation of national health insurance. AB - Since January 1, 2008, the Belgian national health insurance (INAMI/RIZIV) edited a new agreement for the prolongation of the pilot-study on spinal cord stimulation for chronic critical unreconstructable lower limb ischemia. After a short introduction and a summary of the results of the initial Belgian pilot study (2000-2005) on spinal cord stimulation, the official new text is now published in both languages. PMID- 18411594 TI - Current limitations to the histopathological diagnosis of some frequently encountered bone tumours. AB - The final diagnosis of a bone tumour comes in many cases like the last piece of a puzzle which requires integration of clinical, imaging and pathological data. However there are situations in which a discrepancy exists between histology and imaging studies and where histology alone cannot be decisive. This paper reviews such situations. PMID- 18411595 TI - Outcome of cemented Neer II hemiarthroplasty in displaced humeral head fractures. AB - The functional and radiological outcome of cemented Neer II hemiarthroplasty performed within six weeks after a fracture-dislocation or a three/four-part humeral head fracture was evaluated in 48 patients with an average age of 73 years (range: 45 to 89), with an average follow-up period of 44 months (range: 6 to 106). Thirty-two (67%) patients were satisfied or very satisfied. Twenty-nine (61%) patients had no or mild pain, 17 (35%) had moderate pain and two (4%) had severe pain. Constant Score averaged 49 (range: 8 to 71) compared to 78 (range: 40 to 100) for the nonoperated shoulder (p < 0.001). The 26 (57%) patients with abnormal radiographs (malpositioning or loosening of the prosthesis, non-union of tuberosities, heterotopic ossification, and/or glenoid erosion) had a significantly lower Constant Score (45 [range: 8 to 69] versus 55 [range: 31 to 71], p = 0.013). The functional outcome is disappointing and related to the radiographic status. The patients however seem to be satisfied despite a rather stiff shoulder and in our opinion a non-acceptable pain relief. PMID- 18411596 TI - Helix wire osteosynthesis of proximal humerus fractures: unacceptably high rate of failure. AB - The aim of this study was to assess the success rate of osteosynthesis of proximal humerus fractures with titanium helix wires. We retrospectively reviewed the charts of 37 patients who underwent fixation with a helix wire after fracture of the proximal humerus from June 2003 till June 2006. Three patients were excluded because of insufficient details of admission and follow-up. Median age was 64 years (range: 22 to 89). Fractures were classified according to Neer's classification: there were two 3-fragment fractures, thirty 2-fragment fractures and two undisplaced fractures. Internal fixation was performed on average 2.5 days after trauma. During operation either one (18) or two (16) helix wires were used depending on fracture stability. Postoperative immobilisation included approximately 2 weeks in a fixed arm sling and 2 weeks in a collar and cuff. First follow-up was at an average of 2.3 weeks and final follow-up at 14.8 weeks. Follow-up included both radiological and clinical assessment. Already at first follow-up 7 (20%) fractures were displaced, and further surgery was indicated. At final follow-up 9 more fractures were labelled as failures. There was an overall failure rate of 47%. Although helix wire osteosynthesis is a less invasive surgical technique, a failure rate of 47% overshadows the effectiveness of this method. We do not recommend this method for treatment of proximal humeral fractures. PMID- 18411597 TI - Evaluation of functional outcome and patient satisfaction after arthroscopic elbow arthrolysis. AB - Arthroscopic arthrolysis is indicated for stiffness and pain caused by intrinsic stiffness and early arthritis of the elbow joint. Previous studies have demonstrated the benefits in relieving pain and improving motion, but none have reported the specific functional recovery. To understand the functional outcome and patient satisfaction, 26 patients were reviewed at a mean follow-up of 25 months. All were manual workers or strength athletes. Pre- and post-operative evaluation included the Elbow Functional Assessment score, patient satisfaction and return to work and sports. Function improved significantly in 87% and the overall Elbow Functional Assessment score raised from a preoperative 48 to a postoperative 84. Arthroscopic arthrolysis not only improved pain and the range of motion, but also restored the elbow function and returned patients to their desired level of activity. PMID- 18411598 TI - Treatment of scaphoid non-unions with a corticocancellous graft and Herbert screw fixation: results at five years follow-up. AB - In a retrospective survey, 16 patients with 18 established nonunions of the scaphoid treated with a sandwich graft and Herbert screw fixation, were reviewed: 14 patients were very satisfied with the outcome. The mean DASH score was 17 (SD 18.4). Flexion was 76%, extension 79% and gripping force 85% of the contralateral side. Union was achieved in 13 scaphoid bones. There was a significant increase in carpal height. Despite these findings there was also an increase in osteoarthritis. However this was not related to the correction or absence of correction of the preoperatively existing DISI. There was no correlation between radiological presence of osteoarthritis and clinical outcome. PMID- 18411599 TI - Mid-term results of concomitant epiphyseal fixation and trochanteric osteotomy for severe chronic slipped capital femoral epiphysis. AB - The management of severe forms of slipped capital femoral epiphysis (SCFE) has been the subject of intense debate in the literature, and controversy remains as to whether the proximal femoral epiphysis should be realigned by intracapsular or extracapsular osteotomies or just fixated in situ. The aim of this study is to evaluate the late results of treatment of severe unreduced slipped capital femoral epiphyses by combined epiphyseal stabilisation in situ using a single cancellous screw and biplane corrective trochanteric osteotomy. Eighteen hips with severe chronic slipped capital femoral epiphysis before physeal closure were treated by combined epiphyseal fixation in situ using a single cancellous screw, and biplane corrective osteotomy fixed by an angled blade plate. The average follow-up period was 8 years. All patients achieved near-normal hip flexion, internal rotation and abduction, and most were able to bear weight in the early postoperative period. A satisfactory correction of the head-shaft angle was obtained post-operatively on both antero-posterior and frog leg lateral radiographs. There was no instance of chondrolysis, avascular necrosis or early osteoarthrosis. This procedure appears to offer a workable solution to the problem posed by the severely slipped capital femoral epiphysis. PMID- 18411600 TI - The percutaneous compression plate versus the dynamic hip screw: a meta-analysis. AB - Intertrochanteric fractures are a significant orthopaedic burden. The aim of this study was to assess how the Percutaneous Compression Plate (PCCP) technique performs compared to the traditional dynamic hip screw (DHS). A meta-analysis of all head to head trials (1995-2006) comparing the two techniques was performed. Early mortality (< or = 1 year) was the major outcome of interest. Secondary outcomes of interest included operation time (minutes), blood transfusion requirements, post-operative infection and length of stay in hospital (days). There was a decreased trend in overall mortality in the PCCP group [CI 0.84, (0.48 to 1.47), Chi-square = 1.36, p = 0.51]. Similar trends favouring the PCCP technique were seen with the other outcomes. PCCP being a relatively new technique has the potential to become the gold standard in the repair of intertrochanteric hip fractures. However, owing to the limitations of this meta analysis, a large randomised controlled trial is required. PMID- 18411601 TI - Achieving the required medial offset and limb length in total hip arthroplasty. AB - The magnitude of the medial offset and limb length discrepancy after a total hip arthroplasty (THA) significantly affects the biomechanics of the hip. If both of these components are not properly restored, the rate of dislocation may increase. In addition limb length inequality can be a cause for legal problems. We have used a method of intraoperative assessment to restore both the length and the medial offset, and assessed this by comparing the medial offset and leg length in the pre- and post-operative radiographs in 39 consecutive THAs. The median medial offset was 93.9% (range: 85 to 100) preoperatively and 94.2% (range: 85 to 110) postoperatively, compared with the unaffected contralateral side. The median limb length discrepancy was improved from a preoperative -4.84 mm (range: 0 to -30) to a postoperative -0.06 mm (range: -9 to +16). In conclusion, this technique is a simple, accurate and reliable way of restoring the medial femoral offset and correcting the limb length inequality. PMID- 18411602 TI - Total hip replacement for acute femoral neck fracture: a survey of National Joint Registries. AB - Joint registers audit the performance of different types of prosthesis. The data from these registers is invaluable, as the conclusions are evidence based and are not based on the results from a small clinical trial or a case series. The 3rd National Joint Registry Annual Clinical Report (2005) of England and Wales was compared with the latest available online report of other national joint registers, particularly with reference to the usage of total hip replacement performed for fracture of the neck of the femur. In Sweden, total hip replacement is performed for the management of a fracture of the neck of the femur six times more often than in England and Wales, four times more often than in Australia, twice more often than in Canada. Hip fracture registers could provide us with the much needed clinical evidence that could help us solve the last controversy of this unsolved fracture. PMID- 18411603 TI - Patients are more satisfied than they expected after joint arthroplasty. AB - The aim of this study was to compare the preoperative patients' expectations with their postoperative satisfaction after arthroplasties of the hip and knee, using a visual analogue scale. The comparison was made in a group of 44 patients after 44 primary knee and hip joint arthroplasties. A visual analogue scale (VAS) was used for the assessment of expectation and satisfaction. The mean preoperative expectation VAS was 14.8 (SD: 14.3). The mean patient satisfaction at time of follow up was 13.0 (SD : 21.1). We found no agreement in the preoperative patient's expectation satisfaction versus postoperative satisfaction (p = 0.66). Moreover in our study, the patients expected to be less satisfied than they actually were at follow-up, which is shown with the Bland and Altman method. It appears that patients are not capable of predicting the outcome of the joint arthroplasty, which could be influenced by negative preoperative information on complications and risks. Pain and functional disability are probably the most important factors for the patients' satisfaction after arthroplasty surgery. PMID- 18411604 TI - Experimental investigation of negative pressure intrusion techniques of acetabular cementation in total hip arthroplasty. AB - The main mode of failure of the acetabular component in total hip arthroplasty is aseptic loosening. Successive generations of cementation techniques have evolved to alleviate this problem. This paper evaluates one such method, Negative Pressure Intrusion Cementation. Two groups of machined bovine cancellous bone samples were created; experimental (n = 26) and control (n = 26). The experimental group was cemented using the negative pressure technique and the control group was cemented in the absence of negative pressure. The relative cement intrusion depths were then assessed for each group using MicroCT. These samples were then further machined and tested to failure in torsion to estimate their mechanical properties. Results show mean cement intrusion depth for the negative pressure group to be 8,676 microm and 6,042 microm for the control group (p = 0.078). Mechanical testing revealed a greater mean torque in the negative pressure group (1.6223 Nm versus 1.2063 Nm) (p = 0.095). This work quantifies the effect of negative intraosseous pressure on cement intrusion depth in cancellous bone and for the first time relates this to increased mechanical strength. PMID- 18411605 TI - Accuracy and reproducibility of computer-assisted measurement of polyethylene wear in knee arthroplasty. AB - When using computer-assisted methods to evaluate polyethylene wear in knee arthroplasty (TKA), variations in the inclination of the X-ray beam and lack of standard calibration may affect accuracy and reproducibility. To address these issues, we evaluated the polyethylene thickness of unimplanted specimens of known dimensions using the Imagika software. Radiographs were taken with small controlled variations in the inclination of the X-ray beam. Reproducibility was studied based on triplicate measurement of 132 fluoroscopic images by three observers. Calibration was tested against a reference based on a spherical metal ball with a known diameter. The mean differences between the measured and true values ranged from 0.6 mm to 0.8 mm. The repeatability coefficient revealed a maximum variation of 0.43 mm for the same observer, and 0.39 mm between observers. There were significant differences between the measurements of polyethylene thickness performed using two different calibration methods. The variance of measurements was lower with digitized images than with fluoroscopic images. Imagika was not efficient to measure wear in TKA. PMID- 18411606 TI - Predictors for blood transfusion following total knee arthroplasty: a prospective randomised study. AB - A prospective randomised study was conducted on 121 patients undergoing total knee arthroplasty, to identify factors predicting the need for postoperative transfusion. Patients were additionally distributed into two groups, one of which received local adrenaline into the surgical field before wound closure, while the other served as control. A statistically significant correlation was noted between preoperative haemoglobin level, haematocrit and erythrocyte count and the need for transfusion; haemoglobin level emerged as the main predictor. No correlation was observed with patients' age, sex, body mass index, blood pressure or number of comorbidities. Local administration of adrenaline into the surgical field did not result in any reduction in blood loss, nor did it modify transfusion requirements. PMID- 18411607 TI - An analysis of different types of surgical fixation for avulsion fractures of the anterior tibial spine. AB - This is a retrospective study of 25 patients, looking at the outcome of open reduction and fixation of displaced anterior tibial spine avulsion fractures with absorbable and non-absorbable materials. The mean period of follow-up was 44 months (range, 21 to 88 months). The results of surgery were assessed clinically and radiologically. Outcome was evaluated by using the Lysholm knee scoring system. Anterior cruciate ligament laxity was assessed by using a KT-1000 arthrometer and range of movements with a goniometer. Overall, children did better than adults, but age per se did not appear to affect the final outcome. There was no significant difference between children fixed with absorbable or non absorbable materials. Adults fixed with non-absorbable material had significantly better results than those fixed with absorbable material. Herbert screws had a tendency to migrate into the bone substance, and are best avoided. Protected early mobilisation in a knee brace was found to be safe and helped to regain early range of movements in the knee joint. PMID- 18411608 TI - Extracorporeal shock-wave therapy (ESWT) with a new-generation pneumatic device in the treatment of heel pain. A double blind randomised controlled trial. AB - Although low-energy extracorporeal shock wave therapy (ESWT) is widely used to treat a variety of soft tissue disorders, no precise algorithm has been accepted in clinical management. Furthermore, the clinical use of a new generation pneumatic device has not yet been evaluated. We performed a double blind randomised controlled trial on a group of 25 patients with heel pain from chronic plantar fasciitis, to assess the efficacy of ESWT. The main outcome measure was the patients' subjective assessment of pain by means of a Visual Analog Scale (VAS) and the Roles and Maudsley Score before ESWT, early after treatment and six months later. There appeared to be a significant placebo effect with low-energy ESWT in patients with heel pain, and there was also lack of evidence for the efficacy of ESWT when compared to sham therapy. PMID- 18411609 TI - Mini-open percutaneous transarticular screw fixation for acute and late atlantoaxial instability. AB - The treatment of atlantoaxial instability by means of posterior transarticular screw fixation combined with a Gallie fusion is an established method when direct anterior odontoid screw fixation is not indicated or has failed. In this retrospective study, the results of a modified percutaneous mini-open transarticular C1-C2 screw fixation are presented. Between February 1998 and March 2006, 47 patients with acute or late (after failed conservative treatment) atlantoaxial instability were treated with the modified technique. Their average age was 74.9 years. There were no intraoperative injuries to neural structures or blood vessels; 96.8% of the screws were placed correctly. A revision operation was necessary in one patient because of infection at the graft donor site. No patient experienced a neurological complication. Three patients died during hospitalisation, 6 others later on; 6 could not be traced, leaving thirty-two patients or 68% available for follow-up. The average clinical follow-up was 42 months (range: 12 to 91). The results with respect to the pain and activity status were good or excellent in more than 90% of cases. The radiographic follow up averaged 25 months (range: 12 to 75). Bony fusion was documented in all cases. The modified technique of transarticular screw fixation presented here is a safe and functionally satisfactory method of achieving stabilisation of the atlantoaxial complex. Special cannulated instruments are not required. This mini open transcutaneous technique is an alternative to the conventional open procedure, and reduces operation time as well as blood loss. PMID- 18411610 TI - A simple external fixator for complex finger fractures. AB - Complex fractures of phalangeal bones in the hand are difficult to treat. External fixation is an effective method of treatment particularly when internal fixation is not possible due to comminution and associated soft tissue injury. A variety of commercial fixators are available for the treatment of hand fractures. However, these fixators are costly and need a sufficient degree of expertise and familiarity for their use. We describe a very simple fixator which is stable, lightweight and can be easily constructed with materials readily available in most trauma theatres. PMID- 18411611 TI - Lymphangiomyoma of the shoulder: an exceptional localisation. A case report. AB - Lymphangioleiomyomatosis (LAM) is a rare condition usually occurring in young women of childbearing age. It is characterised by the presence of abnormal smooth muscle cells (LAM cells) in the lungs, lymph nodes, and/or other organs, with a few reports of isolated extrapulmonary cases. We report the case of a 26-yr-old female who presented with a painless shoulder mass. Ultrasonography, computed tomography and magnetic resonance imaging revealed an intermuscular mass with cystic and solid tissue components, measuring 6 cm x 4 cm. The mass was removed surgically after the diagnosis of LAM was made following tru-cut biopsy. This case corresponds to an exceptional localisation of LAM. PMID- 18411612 TI - Radial nerve entrapment in a humeral fracture callus. AB - A patient with a radial nerve palsy which persisted nine months after a diaphyseal fracture of the humerus was referred to our department. Radiographs and ultrasound examination showed that the radial nerve was transected and partially entrapped in the fracture callus. This double injury was confirmed and was repaired during subsequent surgical treatment. We report this rare case of combined primary and secondary nerve lesion. PMID- 18411613 TI - Metastasis from renal cell carcinoma presenting as osteolysis in total hip arthroplasty: a case report. AB - We report a case of a pelvic metastasis from a renal carcinoma in association with a total hip arthroplasty. Mistakenly diagnosing such lesion as a granulomatous osteolytic foreign body reaction due to polyethylene debris may lead to devastating and uncontrollable haemorrhage during revision arthroplasty. PMID- 18411614 TI - Osteosynthesis of a periprosthetic fracture of the proximal femur with the distal femur LISS system. AB - In this case report, we show how it is possible to perform osteosynthesis using minimal invasive techniques instead of conventional methods. In this instance the osteosynthesis was performed on a patient in poor general condition who had presented a periprosthetic fracture of the proximal femur. For the osteosynthesis the Less invasive Stabilization System intended for distal femur fractures was used with success. PMID- 18411615 TI - False aneurysm of the anterior tibial artery following total knee arthroplasty. AB - We report a case of false aneurysm involving a branch of the anterior tibial artery following total knee arthroplasty, which presented as an enlarging swelling over the antero-lateral aspect of the leg. False aneurysms are a rare complication of total knee replacement and most of those that do occur arise from the popliteal artery. The swelling was investigated and diagnosed as a false aneurysm and the patient underwent successful percutaneous embolisation of the feeding vessel. This is currently the preferred treatment for small false aneurysms. This procedure, carried out under local anaesthesia, has a low rate of complications and avoids further surgical exploration. PMID- 18411616 TI - Treatment of congenital pseudarthrosis of the tibia with native bovine BMP: a case report. AB - Bone morphogenetic proteins (BMP) have been shown to induce bone formation and union in long bone defects and nonunions. We report a case of congenital pseudarthrosis of the tibia treated with a composite implant consisting of a biocoral frame, collagen carrier, and native bovine BMP extract. A six-year-old boy had persisting congenital proximal tibial pseudarthrosis despite six prior operations. At surgery, the sclerotic surfaces of both fragments were excised, fixation was performed using Ilizarov's device, and the composite implant and an autograft were applied to the nonunion site. Three months after the operation, radiographs showed union, and at four months, the Ilizarov device was removed. Two years later, the proximal pseudarthrosis remained clinically and radiologically united. It is concluded that BMP may contribute to the healing of congenital tibial pseudarthrosis of the tibia. PMID- 18411617 TI - Non-operative management of a talar body fracture in a skeletally immature patient. AB - We present the case of an 11-year-old girl who presented to our Emergency Department having fallen from a height of 8.5 meters. Amongst the sustained injuries was a comminuted fracture of the body of the talus. This was managed conservatively with an excellent outcome. We discuss the case with reference to the literature in this field, particularly highlighting comparisons between cases from patients with mature bone and those with immature bone. PMID- 18411619 TI - [Research in preservation and renal transplant]. PMID- 18411618 TI - Progression of spondylolysis to isthmic spondylolisthesis in an adult without accompanying disc degeneration: a case report. AB - Progression of spondylolysis to spondylolisthesis in adults is very rare. It is always accompanied by disc degeneration at the slip level, or at a lower level. The intervertebral disc is indeed the main structure that opposes the anteriorly directed shear forces. Of course, the disc degeneration might also be a consequence, rather than a cause of the slip. The authors describe an unusual case of progression of spondylolysis to spondylolisthesis in an adult, without any disc degeneration. They are not aware of a similar case in the literature. In 1999, an aircraft engineer with known asymptomatic spondylolysis was involved in a low impact motorcycle accident, after which a Grade I spondylolisthesis L4 was diagnosed. There was no predisposing disc space narrowing at any vertebral level. There may have been a certain degree of microscopic disc degeneration L4L5, a possibility which was confirmed by the development of a disc hernia L4L5, seven years after trauma. This case illustrates the potential for progression of spondylolysis to spondylolisthesis in an adult, without radiographical signs of disc degeneration at any level. The minimal trauma might have played a role. The authors recommend that patients with known spondylolysis who sustain acute exacerbation of their back pain should have standing radiographs. PMID- 18411620 TI - [Research and surgery]. AB - From the Second World War onwards an amazing development in science and clinical & surgery practice has taken place: antibiotics, blood storage, cardiac surgery, organ and tissue transplant, complete joint replacement, total intravenous nutrition or minimally invasive surgery. Paradoxically, during the last two decades of the XXth century, social popularity of doctors has decreased at the same time of some important changes: increases in costs of medical attention, trials against medical mistakes, some doubts on the real role of Academic Public Hospitals and rising in the importance of alternative medicine. Increasing complexity of biomedical research in the continuous changing age of molecular biology has promoted an increasing scepticism regarding clinics and surgeons are able to keep on contributing to medical advances. The study of the contemporaneous History of Medicine demonstrates that some of the more significant achievements have been accomplished by surgeons. Undoubtedly Science and Clinics must get adapted to these times of change and persist in generating important findings. Current Clinical and surgical practice is completely determined by yesterdays' scientific research; tomorrow won't be different. PMID- 18411621 TI - [Experimental renal preservation units: is this activity necessary in Spain?]. AB - Renal graft preservation research is considered complementary but at the same time important activity in high-quality Kidney Transplant Programs. It provides information on limitations of kidneys to be transplanted and to discard those organs with high probability of failure. Unification of criteria in the selection of organs is essential. We think that this activity should be encouraged by Health Institutions although requires investment in staff and technology. PMID- 18411622 TI - [Design of experimental models in surgical investigation]. AB - INTRODUCTION: Research in Medicine is essentially based on three knowledge resources: diseased people (natural and primary), cadaveric bodies (Pathology primary resource) and experimental animals, whom constitutes physiopathologic knowledge resource. Experimental advances reached in the last century have determined the change of the concept "experimental animal" to a wider term: "experimental model". OBJECTIVES: This paper tryes to clarify this concept. To attain this goal, we must define previous considerations in the meaning of the concept "experimental models" and its two key-elements: statistics and design of experiments. PMID- 18411623 TI - [Cell energetic loading in experimental renal transplant with different periods of warm ischemia]. AB - INTRODUCTION AND GOALS: Renal procurement after a period of heart st op demands a previous knowledge of ischemia-reperfusion injuries means. To study cell injury mechanisms an experimental study has been designed in pigs, with different rangres of warm ischemia (0-30-45 and 90 min). The main goal was to research on the basis of ischemic injury. MATERIAL AND METHODS: Biochemical parameters (creatinine, urine output), energetic loading (ATP, ADP, AMP and global energetic loading) and pathological studies as long as survival analysis by 5th day were completed. RESULTS: Animal survival and graft viability range from 100% at 5th day in control and 30 min warm ischemia groups to 60% in 90 min warm ischemia group. Creatinine levels rises at 1st, 3rd and 5th day, especially in those non viable organs. ATP levels decrease after warm ischemia period, increases ADP and AMP levels after reperfusion in those viable organs. CONCLUSIONS: Prolonged periods of warm ischemia do not result necessarily in non-viable kidneys. Viable organs recover nucleotide levels early. Study of energetic cell loading levels is a good way to get on better in the knowledge of injury mechanisms after ischemia reperfusion.